PMID- 10452034 TI - [Evaluation of the growth and contraction of composite skin]. AB - OBJECTIVE: To evaluate the growth and contraction of different composite skin grafts. METHODS: The status of the following composite skin grafts was evaluated over 2 years: allogeneic sucking pig dermis and autologous epidermis (group 1, n = 19), allogeneic young pig dermis and autologous epidermis (group 2, n = 7), autologous epidermis (group 3, n = 8). The effects of gestation on the composite skin grafts were also assessed. RESULTS: It showed that composite skin grafts had smooth surface and no scar formation during 2 to 124 weeks after transplantation. After week 36, the surface area of each group enlarged with different growth rates: group 1 > group 2 > group 3 (P < 0.01). Whole piece grafting was significantly better than joining together of smaller pieces after week 24 (P < 0.001). During pregnancy, CS had good growth ability and elasticity (P < 0.01). Histological study showed that group 1 had rapid growth of skin papillae and epidermal ridges similar to normal skin at week 24. CONCLUSION: As CS graft with trypsin treated allogeneic sucking pig dermis as a substitute for dermis showed the advantages of rapid growth, good elasticity and stability, it is suggested that it could be used in clinical practice. PMID- 10452035 TI - [Advances in the experimental study of fetal plastic surgery]. PMID- 10452036 TI - [Repairing of serious facial deformities with osseointegrated implant supporting fixed prosthesis]. PMID- 10452037 TI - [Experimental study of temporal expression of endogenous TGF alpha in partial thickness scalded wounds]. AB - Investigating spatial and temporal expression and distribution of endogenous TGF alpha, and clarifying its role in burn wound healing. Immunohistochemical and mRNA dot blot hybridization approaches were adopted. Thermal injury can induce TGF alpha gene expression. The expression peaks at the time when repair cells migrate and proliferate most actively, and declines on near completion of epithelialization. Otherwise, the mRNA expression of EGF receptor manifests the same change as TGF alpha's. Immunohistochemical investigation indicates that TGF alpha is mainly distributed in the survived hair follicles and reepithelialized cell layers, and the immunoreactive staining is proportional to epidermal cell growth and migration. Thermal injury can induce endogenous TGF alpha and EGF receptor expression, which is well controlled and correlated with the migration and proliferation of keratinocytes. Endogenous TGF alpha plays a positive role in regulation of wound healing. PMID- 10452038 TI - [Experimental study on immunity and burn wound healing]. AB - The immune system is involved in the complicated process of wound healing. In this study, we at first set up an optimal immuno-suppression model with low-doze radiation with 60Co and found that both the ratio of Th/Ts in peripheral blood and the rate of T cells adhesion to tumour cells dropped persistently, and there were no significant injuries to skin tissue DNA and cells cycle in 20 days after 2Gy radiation with 60Co. Then we investigated the influence of immuno-suppression on burn wound healing. The results showed that both ability of collagen repair and rates of wound healing after burn injury were inhibited in immuno-suppression rats. This indicated that it was necessary to improve immunity condition to enhance wound healing after severe burn injury. PMID- 10452040 TI - [Endoscopic subperiosteal facial rejuvenation]. AB - It is just recently that video endoscopic techniques began to be applied to facial rejuvenation in China. Since July 1995, the authors have performed 15 operations of subperiosteal rejuvenation under the video endoscope, combined with liposuction of the sub-mandible region and the nasolabial fold. The operation uses small incisions, produces satisfactory results with less edema, less numbness and without alopecia. The indications, instrumentation, surgical technique and results and presented and discussed. According to our clinical experience, the best candidates for this operation are those aged from 40 to 55. For the patients about sixty years old, a mini multiplane facelift should be carried out in the cheek and neck region following the subperiosteal facelift. The more elderly patients are not candidates for this operation. We emphasize that the dissected scalp flap should be uplifted to the maximum and anchored on the cranial bone to ensure a satisfactory result. PMID- 10452039 TI - [Effects of locally used SD zinc, SD silver and skin transplantation on the combined radiation-burn injury in the rat]. AB - Effects of local use of sulfadiazine (SD) zinc, SD silver in the wound and skin transplantation on the combined injury of 6 Gy radiation and 15% TBSA burn were investigated in the present study. In the three groups of SD zinc or SD silver plus skin grafting at 15 d after combined injury and SD zinc alone, the 15-day survival rates were 63%, 69% and 75% respectively, and significantly higher than that of control group (42%). The repair time of burn wound was shorter and the number of white blood cells was lower than that of control. Animals in the group of local use of SD silver followed by skin grafting on the 7th day all died, but all of the animals survived when the skin grafting was performed after 15 days. These results suggested that SD zinc and SD silver showed some therapeutic effects on the combined radiation-burn injury. In critical phase of the combined injury, surgical operation should be prohibited. However, in the recovery phase, skin transplantation could be performed. PMID- 10452041 TI - [Reconstruction of the skin structure of the aging]. AB - Based on the character of degeneration and change of the aging skin, reconstruction and repair of the skin structure are performed with a method of subdermal incision in order to increase the content of collagen protein and replace the degenerates collagen in the dermis. The animal experiment shows that the treated skin contracts by nearly thirty percent (P < 0.01) at 15 days after surgery. The epidermis becomes thicker and the dermis contains more fiberoblasts and collagen than that of preoperation. Seven patients were treated with the method. Afterwards their crow's feet deformity and the wrinkles of the nasolabial groove almost disappeared. PMID- 10452042 TI - [Craniofacial growth changes following premature closure of the bilateral coronal sutures in rabbits]. AB - The study was designed to examine the amount and the direction of craniofacial growth changes following bilateral coronal sutures (BCS) closure in young rabbits. At 2 weeks of age, the BCS were fused by the application of enamel adhesive. These animals and a control group also received amalgam implants on the frontonasal, coronal, and lambdoid sutures to monitor suture growth. At 4 or 8 weeks after suture fusion, separate groups of animals underwent release of fused sutures. Immobilization of BCS resulted in growth cessation of the coronal suture. The normal sutures underwent very rapid growth from 2 to 8 weeks of age. BCS release at 8 weeks after fusion resulted in growth restriction. Significant compensatory growth was also noted in the frontal and the lambdoid sutures from 2 to 8 weeks. Nasal bone length was increased significantly. Vault length and craniofacial length were reduced. The results of the present study give experimental documentation to emphasize that surgical correction of craniosynostosis should be performed at a young age to achieve better functional and cosmetic results. PMID- 10452043 TI - [Anatomic study of the facial nerve in parotidectomy]. AB - To explore the course and anatomic characteristics of the extra-cranial section of the facial nerve, the authors carried out this clinical study in 120 patients undergoing parotidectomy by dissecting and measuring the facial nerve and its branches regarding its length, diameter and numbers. The results confirmed that the temporal branch is deeper and has multiple branches; the zygomatic branch is bigger, with consistent location and fewer branches; the buccal branch exhibits three patterns: 1. the upper and lower branches, 2. one trunk and 3. one trunk with two or three branches; the marginal mandibular branch is a small and superficial with two to three branches. The authors discuss how to locate the facial nerve accurately. PMID- 10452044 TI - [Experimental study of prostacyclin on the survival of random skin flaps]. AB - The purpose of this experiment was to observe the effect of PGI2 on the survival of the random skin flap. White rats used in the experiment were divided into preoperation (prevention) group, postoperation (treatment) group and the control group. A super-long random skin flap was created on the back of the animal. Postoperatively, the treatment group received intravenous injection of PGI2 while the control group received intravenous injection of normal saline. The blood supply, metabolism and hemorheology of the flap were analyzed. Results showed that the relative blood flow on the distal part of the flap was 41.3% in the treatment group and 22.4% in the control group. The survival rate of the flap of the treatment group was higher than that of the control. There were no differences between the prevention and the control groups. It is concluded that early postoperative injection of PGI2 could improve blood supply and rheology of the flap and increase its survival rate. PMID- 10452045 TI - [Repairing bone and soft tissue defects of the forearm with a composite segmented fibula osteoseptocutaneous flap]. AB - The segmented-fibula osteoseptocutaneous flap has been used to repair the bone and soft tissue defects of the forearm. During the operation, the fibula is divided subperiosteally at proper site according to the bony defects. The two segments of the fibula were then inserted to the radial and ulnar defects before vascular anastomosis is performed. Experience shows that anastomosis of only one group of vessels can ensure survival of both the segmented fibula and the cutaneous flap. Care should be taken to protect the muscle cuff and periosteum of the fibula, which is the source of blood supply to one segment. Four patients with bone and soft tissue defects of the forearm have been treated using this method. All the flaps survived with good function. The success of this technique contributes to the application of the free fibular flap in repair of forearm injury or osteomyelitis. PMID- 10452046 TI - [Local administration of drug during tissue expansion]. AB - In order to shorten the expansion time and reduce complications in tissue expansion, we developed a new method using local delivery of drug during tissue expansion. The procedure commenced with placing an implant syringe between the skin and the expander. While the skin was expanded, the drugs such as prednisolone and procaine were injected under the skin from the implant syringe. We have used the method in 23 cases and compared the results with the control group of 11 cases. It was found that the new method shortened the expansion time and reduced complications. The method had definite effect and the manipulation was simple and convenient. We conclude that the method is of value to assist tissue expansion and is worth employing routinely. PMID- 10452047 TI - [Serum neopterin levels after extensive burns and their relationship to endotoxemia and sepsis]. AB - The present study was performed to determine the kinetics of serum neopterin levels after major thermal injury and their relationship to endotoxemia and sepsis. This prospective study included 35 patients (32 males and three females) with total burn surface area (TBSA) greater than 30% (30%-98%), and 22 healthy volunteers who served as normal controls. The results showed that neopterin levels increased in most patients on day 3 postburn, but they were not significantly correlated with the extent of the burn surface (P > 0.05). Patients with endotoxemia had much higher neopterin values than those who showed no endotoxemia from the second week onward (P < 0.05-0.01), and circulating endotoxin and neopterin levels were positively correlated in patients who developed endotoxemia on day 14 (r = 0.368, P < 0.05) and day 21 (r = 0.439, P < 0.01) after major burns. Moreover, a high serum neopterin level was found in patients with sepsis (n = 15), and the marked elevation persisted throughout the observation period. The difference between septic and non-septic patients (n = 20) became significant on 14 and 28 days postburn. These data suggest that extensive burns can lead to an elevation of serum neopterin independent of TBSA. The endotoxin release in the circulation may be involved in the continuous formation of neopterin during the late postburn stage. In addition, the presence of a constant high neopterin level is associated with a critical event in the development of burn sepsis. PMID- 10452048 TI - [The changes and clinical significance of serum CRP, C3, Tf, and PA in the early postburn stage]. AB - The aim of this study was to investigate the regulation and clinical significance of serum acute phase proteins. The patients were divided into three groups: group I consisted of 11 patients with less than 20% of TBSA; group II 10 patients with 20-50% of TBSA and group III 11 patients with more than 50% of TBSA. Blood samples were taken at 8, 24, 48 and 72 h postburn, and in group III additional blood samples were taken on the 7th, 14th, 21st day or in septic episode. The concentration of CRP and C3, Tf, PA were detected by rate nephelometry, agar immunodiffusion and rocket immunoelectrophoresis respectively. Compared with controls, the peak value of CRP was observed during 48-72 h (P < 0.01), C3 decreased at 8 h and lowest at 24 h. Tf was lowered at 48 h, PA and Tf shared the similar changes. However, all four variables had no marked difference during infection and sepsis. The results showed no correlation between C3 and CRP (r = 0.0885, P > 0.05). The results suggested that: 1. CRP mainly reflected the extent of injury but not sepsis; 2. increased CRP was not the causative factor of decreased C3. Tf reduction accelerated bacterial growth. PMID- 10452049 TI - [Identification of the properties of haptoglobin-like substance in burn serum]. AB - In our previous study, a haptoglobin-like substance possessing immunosuppressive activity was found in the sera of burned patients. However, its properties have not yet been clarified. In the present study, we mainly investigated the properties of the haptoglobin-like substance. The results showed that the abnormal protein bands identified by SDS-PAGE were the different subunits of haptoglobin, i.e., beta, alpha 2 and alpha 1 chains. The haptoglobin in burned sera was identical to that in normal sera in molecular weight, isoelectric point, hemoglobin-binding capacity and immunochemical properties as analyzed by SDS PAGE, isoelectric focusing, immunochemical and biochemical methods. According to our results, it is suggested that the haptoglobin-like substance reported in the past is probably the haptoglobin, and that there may be no alteration in the properties of haptoglobin following burns. PMID- 10452050 TI - [Levels of serum G-CSF and supplement of GM-CSF in rats sustaining delayed resuscitation after major burn]. AB - Delayed fluid resuscitation of burn shock may lead to infection in early period following major burn injury, resulting in multiple organ failure with high mortality. The change in colony stimulation factors (CSFs) may play a role in developing infection. We assessed the levels of serum granulocyte colony stimulation factor (G-CSF), tumor necrosis factor-alpha (TNF-alpha), phagocytosis of neutrophil and counts of peripheral blood cells of rats with 30% TBSA full thickness burn having either immediate or delayed fluid replacement. We also evaluated the efficacy of granulocyte-macrophage colony-stimulating factor (GM CSF) in improving survival of rats following burn injury with a superimposed burn wound infection. THE RESULTS: 1. A delayed rise in serum G-CSF was found in delayed resuscitation group, and the levels of serum G-CSF and phagocytosis of neutrophil were lower compared with immediate resuscitation group. 2. Nonsurvival group had lower levels of serum G-CSF and higher content of TNF-alpha compared with survival. 3. Supplement of GM-CSF could significantly improve animal survival with burn wound infection following severe burn shock. CONCLUSION: Decrease in G-CSF production plays a role in developing fatal wound infection after severe burn shock; supplement of GM-CSF is beneficial in improving survival. PMID- 10452051 TI - [Effects of CD11/CD18 on adhesion between endothelial cell and PMN and its expression in early stage postburn]. AB - This study was to investigate the expressions of CD11a/CD18 and CD11b/CD18 on burn patient's polymorphonuclear neutrophil (PMN) in early postburn stage and effects of CD11/CD18 mAb on PMN-EC adhesion and endothelial cell (EC) damage induced by PMN. Adhesive rate between PMN and EC, the filtration coefficient (kf) and monolayer volume flux (Jv) were determined after incubating burn patient's PMN with human umbilical vein endothelial cell (HUVEC) for 24 hours. CD11a/CD18 monoclonal antibodies (mAb) and CD11b/CD18 mAb were used to pretreat burn patient's PMNs in order to demonstrate the role of CD11/CD18 in PMN-EC adhesion and EC damage. Expressions of CD11a/CD18 and CD11b/CD18 were determined by flow cytometry 1, 3, 5, 7 d postburn. The results indicated that expressions of CD11a/CD18 and CD11b/CD18 on burn patient's PMNs increased quickly, reaching the peak on day 1 postburn and remained in high level till 7 days postburn. The values of Kf and Jv were increased markedly after burn patient's PMNs was incubated with HUVECs, which were cultured on micropore filter membrane. When burn patient's PMNs were pretreated with CD11a/CD18 mAb and CD11b/CD18 mAb, PMN HUVEC adhesive rate was inhibited by 70%-80% and the values of Jv and kf were decreased significantly. It suggested that there were increased expressions of CD11a/CD18 and CD11b/CD18 on PMNs after burn injury. Burn patient's PMNs could damage EC and increased EC monolayer permeability (kf, Jv). CD11/CD18 mAb inhibited PMN-EC adhesion and attenuated EC damage induced by PMNs. PMID- 10452052 TI - [Analysis of antibiotic resistance and production of enterotoxin of MRSA]. AB - 100 separate Staphylococcus aureus (S. a) isolates at Beijing Jishuitan Hospital during Oct. 1994-Apr. 1995 were submitted to determine the proportion of methicillin-resistant S. a (MRSA) and susceptibility test to 12 antibiotics. MIC determinations of the antibiotics were performed. Also, production rate of enterotoxin was measured with RPLA method. The results showed that MRSA constituted 60% of all cultures. (Burns Dept. 67.4%, Internal Medicine 62.5%, Orthopedics Dept. 44%). The antibiotics with resistant rate of MRSA below 50% included vancomyin (3.3%), norvancomyin (5%) and amikacin (21.5%). The production rate of enterotoxin was 100% in MRSA and 20% in MSSA (methicillin-sensitive S. a). In this group, 35% of MRSA and 5% of MSSA produced more than 2 types of enterotoxin. PMID- 10452053 TI - [Subcutaneous vascular network flap transplantation]. PMID- 10452054 TI - [Effects of surgical flaps on wound healing and the elimination of bacteria from wound]. PMID- 10452055 TI - [Clinical application of photodynamic therapy combined with non-coherent light (red light) for treatment of port-wine stains]. AB - OBJECTIVE: This is to report the clinical application of photodynamic therapy (PDT) combined with non-coherent light (red light) for the treatment of port wine stains (PWS). METHODS: Eleven cases with PWS have been treated with PDT combined with non-coherent light since Dec. 1995. RESULTS: The facial lesions of PWS in the 11 cases disappeared completely without any scarring. CONCLUSION: PDT combined with red light that has strong skin-penetrating ability and irradiation just after intravenous injection of photosensitizing drugs are effective to embolize the malformative vasculature in pink lesions, especially in dark purple lesions and purple lesions with proliferation. PMID- 10452056 TI - [Measurement of mandibular angle protrusion and semi-blind amputation of hypertrophied mandibular angle with the help of a pre-made mold]. AB - OBJECTIVE: Mandibular hypertrophy is a subjective diagnosis to some extent. The authors tried to find a way for true objective diagnosis of mandibular hypertrophy. METHODS: Radiographic cephalometry was performed in 39 randomly selected adults. The degree of the mandibular angle was measured and summarized. According to measurement of the mandibular angle and the extent to be amputated, we designed and made various molds. RESULTS: With this pre-made mold, we amputated semi-blindly and exactly the desirable size and shape of the mandibular angle. This technique has been used in 6 patients for mandibular amputation and masseter thinning. In three patients shaving of the out layer of the hypertrophied mandible was also done. CONCLUSION: The new technique using the pre made operative mold shortens operating time, reduces surgical bleeding and tissue swelling, and can help to remove the bony block precisely. PMID- 10452057 TI - [Correction of square face]. AB - OBJECTIVE: This work was to provide an advanced operation method for square face correction. METHODS: The method is characterized by resecting hyperostosis of the mandibular angle, the bulky masseter and the buccal fat pad through intraoral or combined extra-intraoral approach. RESULTS: Good results were obtained in 10 cases after follow-up of 4 to 28 months. CONCLUSION: This operative method proved to be quite ideal and applicable to treat square face. PMID- 10452058 TI - [Multiple-layer composite rhytidectomy]. AB - OBJECTIVE: The multiple-layer composite rhytidectomy is presented in this paper for correction of the facial aging. METHODS: The operative procedures include multiple-layer dissection, suspension reconstruction of the zygomatic ligament, and thinning of the zygoma. RESULTS: The method has produced a good result in 28 patients since 1993. CONCLUSION: Study of clinical cases and cadaver dissections had led to better understanding of the aging face. With advancing age, all tissues of the face lose their balanced relationship. These changes occur in their volume shape, position, and consistency. With the multiple-layer dissection and suspension, the soft tissues of the check, forehead, nasolabial fold, lateral canthus and eyebrow can be lifted to reestablish their previously youthful relationship with the underlying skeleton. PMID- 10452059 TI - [Applied anatomy of the temporal and frontal regions in endoscopic temporal lifting]. AB - OBJECTIVE: This anatomic study was carried out in a bid to decrease damage of the temporal branch of the face nerve in endoscopic temporal lifting. METHODS: We dissected the frontal-temporal region and located the temporal branch on 36 sides of adult head specimens. RESULTS: It was found that the temporal branch is deep to the temporoparietal fascia and frontal muscle. The temporal branch has one to four branches in general. The first one is 8.1 +/- 2.1 mm anterior to articular tubercle and 33.8 +/- 4.9 mm superior to the bony lateral canthus. The temporal branches locate below a line drawn from the articular tubercle to 40 mm above the bony lateral canthus. CONCLUSION: It is suggested that the operation incisions in the temporal region be selected above this line. PMID- 10452060 TI - [The pathological action of surrounding connective tissues of alar cartilage in the cleft lip nose]. AB - OBJECTIVE: This study was to explore the pathological action of the adjacent connective tissues of alar cartilage in unilateral cleft lip nose. METHODS: Anatomic-histological and pathoanatomical study was carried on 40 normal cadavers and 30 patients with secondary deformities of unilateral cleft lip during their operations. RESULTS: Six kinds of connective tissues with different histological features were found between the alar cartilage and adjacent skin, cartilage or bone. There was no histological difference in each connective tissue between the normal and the cleft lip nose. In the cleft lip nose, due to histological differences among the six connective tissues, there existed various changes of pathological dislocations between the alar cartilage and adjacent skin, cartilage or bone. CONCLUSION: The authors suggest that the surrounding connective tissues of the alar cartilage play an important role and be the intrinsic basis to produce the characteristic deformities of the cleft lip nose. The result is helpful in understanding the anatomical and histological features of the connective tissues around the alar cartilage and the deformities of the unilateral cleft lip nose so as to select a reasonable operation method. PMID- 10452061 TI - [The application of the pedicled buccal fat pad graft in cleft palate repair]. AB - OBJECTIVE: The purpose of this paper is to provide the experience in using the pedicled buccal fat pad (BFP) graft in cleft palate repair when healing by secondary intention may need to be considered due to the width of the cleft; the qualitative or quantitative tissue constraints, the dead cavity between the oral and nasal layers or single layer closure of the hard palate with the vomer and marginal pedicled mucosoperiosteal flap. METHODS: The body and buccal extension of BFP is gently torn out from its bed and advanced into the palate vault around the posterior aspect of the maxilla to cover the raw surface of the hard palate cleft and/or fill the dead cavity between the oral and nasal tissue layers, specifically in the conjunction of the hard and soft palate. RESULTS: Six cleft palates were repaired with the BFP graft. The postoperative course was uneventful. The BFP graft is viable without the occurrence of fistula or infection. CONCLUSION: The pedicled BFP graft has provided an excellent soft tissue support for secondary epithelialization when primary dehiscence has or would have otherwise thwarted ultimate closure of the palatal defect. The pedicled BFP graft permits closure of palatal defects that could not be repaired by conventional procedures. PMID- 10452062 TI - [Techniques in fetal rabbit surgery]. AB - OBJECTIVE: Experimental fetal surgery has been a popular way to study growth and development and wound healing. The rabbit continues to be a reasonable animal model. The key to success of the experiment lies in keeping the survival rate of the operated animal above 80 percent. In this paper a simple experimental method for fetus surgery in rabbits is described. METHODS: Intravenous thiopental sodium is used for general anesthesia. Surgery is performed inside the uterus through hysterotomy or with the caudal part of the fetus out of the uterus. Amniotic fluid is restituted by instillation of warm saline at 37 degrees C. Special care is taken to suture the amnion. RESULTS: Using these techniques carefully, we have had 90 percent of success in fetal surgery experiment. CONCLUSION: A technically simple procedure has been developed for fetal rabbit surgery. PMID- 10452063 TI - [Repair of large and complicated defect with multiple expanders]. AB - OBJECTIVE: A method is introduced to obtain good aesthetic and functional results in repairing complicated defects on the face and neck, or total skin defects of the upper limb. METHODS: Two to four expanders are implanted in different areas where the color and texture of skin are similar to that of the defects, expanded simultaneously. During the second operation, several flaps corresponding to the defect areas are created and employed in one procedure. RESULTS: Six patients of 4-41 years old were treated with this technique. All of them have been followed up for 12-18 months with satisfactory results. CONCLUSION: Carefully planned, this maneuver shows better effects than the conventional method in repairing the skin defects of face, neck and upper limb. PMID- 10452064 TI - [Clinical guidelines for timing of escharectomy and skin grafting during burn shock stage in extensively burned patients]. AB - OBJECTIVE: To provide practical clinical guidelines to doctors who have no hemodynamic monitoring facilities in performing escharectomy during the shock period in extensively burned patients. METHODS: We analyzed our clinical experiences in 60 patients with extensive burn. RESULTS: Puting forward several clinical indexes for timing of escharectomy during burn shock stage: 1. Amount of fluids in the first 24 h postburn 2.6-3.0 ml.kg-1.1% TBSA-1; 2. Output of urine 80-100 ml/h; 3. Mentally fully conscious; 4. Thirst significantly alleviated and there is no nausea and vomiting; 5. Pulse 100/min; 6. Hb < or = 150 g/L; 7. Hct < or = 0.50. CONCLUSION: With the clinical indexes as guidelines, we assume that escharectomy could be performed during burn shock stage with reasonable safety. PMID- 10452065 TI - [Influence of escharectomy during burn shock stage upon the changes in endothelin and NO]. AB - OBJECTIVE: To observe influence of escharectomy during and after burn shock stage upon the changes in plasma ET and NO. METHODS: 35% TBSA full-thickness burn was produced in pigs, which were divided into two groups. Group S was a group undergoing escharectomy during burn shock stage at 24 h postburn. Group C was a group receiving escharectomy at 96 h after injury. ET and NO were determined and ET/NO was calculated. RESULTS: The levels of ET and NO were higher after injury than before injury in both groups. The level of ET was lower significantly in group S than in group C from 96 h postburn on. The level of NO was higher in group S than in group C. The ET/NO was lower in group S than in group C from PBH 24 on. CONCLUSION: Escharectomy during burn shock stage was beneficial in reducing injury of endothelial cells, decreasing exudation and edema, decreasing tissue hypoxia and improving microcirculation. PMID- 10452066 TI - [An analysis of bacterial resistance to antimicrobial agents in a burn centre]. AB - OBJECTIVE: To investigate bacterial resistance to antimicrobial agents in the background of extensive employment of third-generation cephalosporins in the treatment of burn infection. METHODS: Bacterial susceptibility testing was carried out using Kirby-Bauer method. RESULTS: Out of 259 Gram negative bacilli isolates, 31% of these strains were all resistant to cefotaxime, ceftriaxone, cefoperazone and ceftazidime. In vitro test, susceptibility of 52% third generation cephlosporins resistant strains were restored by cefoperazone/sulbactum. 23.8% of P. aeruginosa were resistant to imipenem, 51.2% to ciprofloxacin. 28.6% of A. anitratum were resistant to imipenem, 21.4% to ciprofloxacin. 9.9% of enterobacteriaceae were resistant to imipenem, 20.4% to ciprofloxacin. 26.9% of MRSA were resistant to imipenem, 73% to ciprofloxacin. No norvancomycin resistance was detected. 2.7% of E. faecalis were resistant to norvancomycin. CONCLUSION: It is likely that the observed resistance to third generation cephalosporins may be partially due to chromosome-mediated type-1 beta lactamase. PMID- 10452067 TI - [The effect of certain pro-inflammatory mediators on the pathogenesis of acute gastric mucosal lesion in early burn stage]. AB - OBJECTIVE: To appraise the effect of certain pro-inflammatory mediators on the pathogenesis of acute gastric mucosal lesion in early burn stage. METHODS: 21 patients with burn injuries of over 30% TBSA were divided into A and B groups according to the main clinical indexes during shock resuscitation and hemodynamic parameters. Fiberoptic endoscopic examination, determination of intramucosal pH (pHi) and measurement of some mediators were done immediately after admission to the hospital and 4 and 7 days after burn injury. RESULTS: It was demonstrated that the level of LPS in plasma, the content of TNF-alpha, IL-8 and ET in group B at 4 and 7 days postburn were significantly higher than those of group A, while the value of pHi in group B was markedly lower than that of group A. Damaging index of gastric mucosa was negatively correlated with pHi (r = -0.89, P < 0.05), but positively with ET (r = 0.91, P = 0.05). CONCLUSIONS: These findings suggest that the inflammatory mediators and cytokines promoted secondary damage to gastric mucosa during early postburn. It was believed that pHi was a sensitive index, and it played an important role in the development of stress ulceration. PMID- 10452068 TI - [The dynamic changes and significances of endothelins in early burned rats]. AB - OBJECTIVE: To elucidate the significance of endothelins in visceral injury at the early stage of burn. METHODS: By use of a 30% (III degree) total body surface area (TBSA) burn rat model, we measured the concentration of endothelins in plasma, heart, liver, lung, kidney, and observed the effects of PD145065, a kind of non-selective ETA-ETB receptor antagonist, on visceral impairment. RESULTS: Not only did the endothelin concentration in plasma of burnt rat elevated significantly, showing a peak at 6 hours postburn, but also the endothelin contents in internal organs apparently increased. The changes in endothelin in liver or kidney were negatively correlated with the alteration in renal or hepatic blood flow. Moreover, administration of endothelin receptor antagonist PD145065, without affecting the arterial blood pressure, could revert the decrease of renal or hepatic blood flow to some extent, reduce the water contents of heart, liver, lung, kidney postburn. CONCLUSION: The abnormal changes in endothelins postburn contribute to visceral injury in the early postburn stage in rats. PMID- 10452069 TI - [Myocardial contractile and calcium transport function after severe burn injury]. AB - OBJECTIVE: This study examined the changes in myocardial calcium transport function and what roles it played in cardiac dysfunction after severe burn injury. METHODS: Contraction relaxation properties of the left ventricule (LV) were studied in isolated heart preparations of Wistar rat 3, 8, 24 hours after 30% TBSA full thickness burn. Sarcoplasmic reticulum (SR) calcium transport function was measured by millipore filtration technique. RESULTS: The maximal rates of LV pressure elevation (+ dp/ dtmax) and depression (-dp/dtmax) in burn group were significantly lower than that in control group (P < 0.01). The initial rate and capacity of calcium uptake by SR decreased markedly in burn group than in control group (P < 0.01). In addition, calcium dependent ATPase activity and coupling ratio of SR were also markedly depressed. CONCLUSION: SR calcium transport function depression was closely related to cardiac contractile dysfunction, and the former is one of the important mechanism for cardiac contractile dysfunction after severe burn injury. PMID- 10452070 TI - [Protective role of endogenous nitric oxide to microcirculation of rats during burn shock]. AB - OBJECTIVE: To investigate the role of nitric oxide (NO) in burn shock. METHODS: A model of 40% TBSA deep II burn injury in rats was used. The effects of L-NAME, the inhibitor of NO synthase, and L-Arg, the precursor of NO synthesis, on the mean arterial pressure (MAP), microcirculation of muscle, concentration of NO2( )/NO3(-) in plasma and the survival time of animals were observed. RESULTS: The results showed that the synthesis of nitric oxide was increased significantly, accompanied with the decrease of MAP and the disturbances of microcirculation of muscle after burn. L-NAME inhibited the decrease of MAP, but aggravated the disturbances of microcirculation and shortened the survival time of animals, while L-Arg produced the contrary effects. CONCLUSION: The results indicated that the large amount of NO in blood after burn may play an early protective role to injury. PMID- 10452071 TI - [Introduction to tissue engineering]. PMID- 10452072 TI - [Post-trauma stress and enteral nutrition]. PMID- 10452073 TI - [Radioimmunoassay of substance P in human hypertrophic scar]. AB - OBJECTIVE: To measure the content of substance P in human hypertrophic scar. METHOD: The method of radioimmunoassay was used to determine the content of substance P in human hypertrophic scar, non-hypertrophic scar and normal skin. RESULT: The content of substance P in hypertrophic scar was significantly higher than that in non-hypertrophic scar and normal skin (P < 0.01). CONCLUSION: Substance P may play an important role in the process of hypertrophic scar development. PMID- 10452074 TI - [Fibronectin mRNA expression in hypertrophic scars]. AB - OBJECTIVE: This study was to compare the content of fibronectin messenger RNA in the normal skin and hypertrophic scars. METHODS: The mRNA levels of fibronectin were measured by quantitative RT-PCR. RESULT: The study revealed that in vivo, the content of fibronectin messenger RNA was higher in hypertrophic scars than in normal skin. CONCLUSION: It indicates that altered regulation of extracellular matrix components is an important factor for abnormal human wound healing. PMID- 10452075 TI - [Study on in situ expression of type I and III procollagen mRNAs in different parts of keloid and normal skin]. AB - OBJECTIVE: Based on the histomorphologic differences of three parts of keloid, i.e., invasive, proliferative and senile regions, the study was carried out to explore the differences of expression of type I and type III procollagen mRNAs in the three different parts of keloid and normal skin. METHOD: Four samples of keloid from four patients and two samples of normal skin were collected. The expression of pro alpha 1 (I) and pro alpha 1 (III) procollagen mRNAs in keloid and normal skin was investigated by in situ hybridization techniques. RESULT: Both type I and III procollagen mRNAs levels increased in keloid, especially type I, resulting in elevated ratios of type I to III procollagen mRNA. In keloid, expressive intention of type I and type III procollagen mRNAs was higher in the papillary layer than in the reticular layer. Also the expressive intention of type I and type III in keloid was higher in invasive and proliferative regions than in the senile region. However, expressive intention of type I and type III procollagen mRNAs in the invasive region of keloid was similar to that in the proliferative region. CONCLUSION: The differences of expression of type I and type III procollagen mRNAs in different pathological regions of keloid and normal skin is one cause of forming different pathological regions in keloid. PMID- 10452076 TI - [Comparative study on biological characteristics and ultrastructure of the fibroblasts derived from normal skin, hypertrophic scar and keloidin in vitro culture]. AB - OBJECTIVE: This study was to investigate the in vitro cultivation, biological characteristics and ultrastructure of the fibroblasts derived from normal skin, hypertrophic scar and keloid, to elucidate the value of their application. METHOD: A comparative study was performed on cell proliferation, cell morphology, cytogenetic feature and cell ultrastructure of fibroblasts from normal skin, hypertrophic scar and keloid by means of the technique of in vitro culture. RESULT: The results indicated that the fibroblasts isolated from normal skin, hypertrophic scar and keloid exhibited similar morphology and growth rates. The cytogenetic feature and cell ultrastructure were also alike. CONCLUSION: According to our study and that of others, we conclude that it is entirely feasible to establish an in vitro culture model of fibroblasts from normal skin for investigation, prevention and treatment of scars. PMID- 10452077 TI - [Effects of the serum obtained from smoke inhalation injury rats on PMN adhesion and transmigration in vitro]. AB - OBJECTIVE: This study was conducted to determine the changes in PMN adhesive capacity to endothelial cells and transmigration through endothelial cell (CEC) sheet in the presence of the serum from rats with smoke inhalation injury (SII). The effects of adhesion molecules CD11a, CD11b, and ICAM-1 on the PMN adhesion and transmigration were also studied. METHOD: In the present study fluorescent assay, EC sheet cultured on the 5 microns permeable membrane and antibody blocking technique were used to study the PMN adhesion and transmigration in vitro. RESULT: It was found that the serum from SII rats could promote PMN adhesion to CEC sheet. After being cultured with the serum for 12-24 h, PMN adhesion to CEC was more than three times of that in control. The serum also enhanced PMN transmigration through the CEC sheet in vitro, doubling the amount observed in control when the sheet was cultured with the serum for 24 h. Based on the above data it was believed that the SII rat serum could enhance PMN adhesion to and transmigration through lung capillary, and the percentages of PMN adhesion and transmigration were found to be positively correlated (r = 0.975). Furthermore, the antibodies against CD11a, CD11b, and ICAM-1 reduced PMN adhesion 44%, 55% and 51%, respectively, and at the same time decreased PMN transmigration 39%, 65%, and 58%, respectively. CONCLUSION: In the presence of serum obtained from SII rats PMN adhesion to and transmigration through CEC sheet increased, which is the possible underlying mechanism responsible for PMN infiltration in vitro. Moreover, the adhesion molecules such as CDA11a, CD11b, and ICAM-1 play an important role in the process. Infiltrated PMN could release oxygen free radicals and protease to injure lung tissue. PMID- 10452078 TI - [Effects of high frequency jet ventilation on respiratory airflow and gas exchange in dogs with inhalation injury]. AB - OBJECTIVE: The experiment was to investigate the effects of respiratory airflow on gas exchange. METHOD: Eight healthy dogs were anesthetized with thiopentone. A 3-D partial dynamic analyzer was used to determine the inspiratory and expiratory flow velocity, and arterial blood samples were collected for blood-gas analysis during spontaneous respiration. Intravenous muscle relaxant was given quickly and the animals were kept apneic and then vapor (105 degrees C) was inhaled for 5 seconds. HFJV was carried out at random under 5 different conditions, each for 20 minutes. Changes in inspiratory velocity (Vi), expiratory velocity (Ve), Reynole's number, PaCO2, PaO2, and PIP were observed. RESULT: 1. During spontaneous respiration, airflow was in form of laminar flow when Re < 1000, and expiratory airflow was either laminar or turbulent when 1000 < Re < 1500. 2. During HFJV, both expiratory and inspiratory airflow were turbulent when Re > 1500. 3. The expiratory velocity with HFTJV increased significantly (P < 0.05). PaCO2 decreased significantly (P < 0.05) compared with HFJV. Obvious negative correlation was found between them (r = -0.9216, P < 0.05), meanwhile inspiratory velocity had an increasing tendency. CONCLUSION: 1. HFJV could maintain normal ventilation of dogs with inhalation injury. The mechanism might be associated with turbulent flow. 2. On the basis of HFJV, HFTJV increases inverse jet airflow, thus the respiratory airflow velocity may be increased and CO2 elimination improved. PMID- 10452079 TI - [Myocardial function after fluid resuscitation with addition of anisodamine in severely burned rabbits]. AB - OBJECTIVE: To evaluate the effect of fluid resuscitation with addition of anisodamine (Ani) on myocardial function in severely burned rabbits. METHODS: III degree burn injury covering 25% total body surface area (TBSA) was produced on rabbits. LVSP, LV +/- dp/dtmax were measured with fluid resuscitation alone, administration of Ani alone, and fluid resuscitation plus Ani before burn, immediately after burn and 1, 3, 5, 7 h after burn. RESULTS: It was showed that myocardial contractility and relaxation were depressed 1 h after burn. Fluid resuscitation alone and administration of Ani alone could significantly improve myocardial contractility and relaxation only within 3 h postburn. With fluid resuscitation plus Ani (5 mg.kg-1), myocardial contractility and relaxation were improved statistically significantly as compared with control group without treatment (P < 0.01), with the values approaching the normal level. CONCLUSION: Our data demonstrate that the administration of Ani (5 mg.kg-1) in addition to fluid resuscitation is beneficial to burn injury, especially to myocardial function, and it provides a promising approach for the future treatment of burns. PMID- 10452080 TI - [Effects of supplement Zn on levels of Zn in serum, growth hormone and hydroxyproline]. AB - OBJECTIVE: In order to research the effects of supplement Zn on wound healing, the changes of Zn in serum and skin, growth hormone and hydroxyproline of skin were observed in burn rats. METHOD: 80 SD rats were randomly divided into four groups: Control group of 8 rats, N group (diet containing Zn 40 micrograms/g), W group (application of Zn on burn skin), H group (diet containing Zn 80 micrograms/g). 8 rats of each group were killed on 1, 3, 7 days after burn. RESULTS: The levels of serum Zn declined on 1 day postburn in N and W groups, whereas the content of serum Zn in H group elevated and were 2.4, 2.0 times as much as that of N and W groups respectively (P < 0.01). Zn in burn skin increased significantly on 3, 7 days in W group and was 1.3, 1.5 times as much as that of N group. Serum growth hormone increased in all groups postburn. The highest level appeared in H group. Hydroxyproline in burn skin decreased in all groups, but it began to increase on 7 day postburn in W group. CONCLUSION: Supplementation of Zn by diet rapidly raises the levels of Zn in serum, burn skin and growth hormone. It is beneficial to apply Zn to the wound to improve wound healing. Hydroxyproline increased significantly with application of Zn to burn wound. It is better to supply Zn by diet and topical application simultaneously. PMID- 10452081 TI - [Experimental high-voltage electrical burns on limbs of rabbits]. AB - OBJECTIVE: The study was designed to investigate the changes in some rheological parameters and ET-1 in venous blood following electrical burns. METHODS: One of the hind limbs of rabbits was subjected to 3000 V, 2.5 A charge of alternative electric current for 3 seconds. Rheological parameters such as red cell deformability (RED) and platelet aggregation rate (PAR), as well as endothelin-1 (ET-1) in venous blood drained from electrical burn wounds and contents of ATP in damaged muscle were measured at time intervals of 2 h, 24 h, 48 h and 72 h postburn. RESULTS: The changes in the parameters mentioned above were significant following electrical injury. The administration of PGE1 showed an advantageous effect in prevention of thrombosis and muscle "progressive necrosis" through improvement in RCD, reduction of PAR and the content of ET-1 in local circulation. CONCLUSION: RCD, PAR and the content of ET-1 in local circulation changed significantly following electric injury and may play an important role in the process of "progressive necrosis". The potential use of PGE1 in the treatment of electrical injury is proposed. PMID- 10452082 TI - [Restoration of elbow joint and its function by transposition of latissimus dorsi myocutaneous flap following serious high-voltage injury in upper limb]. AB - OBJECTIVE: This is to introduce a method for repairing and restoring the elbow joints and their function of the patients suffering from serious high-voltage electric burns in upper extremity. METHODS: We designed the transposition of latissimus dorsi myocutaneous flap (MCF) to repair the wounds of elbow, as well as restore the joint's flexion function by the motor power of the muscle. This procedure salvaged the elbow joints, which would have to be amputated before. RESULTS: 8 cases, 10 limbs were involved. All flaps survived completely and 6 of them had primary healing while the other 4 flaps healed after dressing changing. The elbow joints and their functions were satisfactorily restored in 5 limbs and the functions were also regained through further elbow joint release procedures in 2 limbs. The other 3 limbs (2 cases) lost contact with us and so the results were not clear. CONCLUSION: This method has evident advantages and satisfactory results. PMID- 10452083 TI - [Clinical characteristics of burn caused by coal mine explosion]. AB - OBJECTIVE: To investigate the characteristics of the burn injury caused by coal mine explosion so as to enhance the cure rate. METHODS: Analyse the therapeutic result after planned standard treatment of clinical patients and review historical patients. RESULTS: 1. Coal mine explosion includes two types, i.e., gas explosion and coal dust explosion. 2. This kind of burn is a combined injury with pathologic changes of burns as the main feature. Mechanical injury is the chief cause of early death. Blast injury mainly damages the lungs. The occurrence of carboxyhemoglobinemia is not often. 3. The amount of fluid infusion in the first 24 h in exudation phase is 8% less of the traditional formula. Alkaline balanced salt solution is supplied as electrolyte solution, which can provide 45% of necessary HCO3- for correction of acidosis. 4. No thorough debridement is imposed in the treatment of burn wound. Baking with electric bulb with topical SD Ag in semi-exposure state can be used. 5. Inhalation injury chiefly occurs in upper respiratory tract. The main bacterial species causing complicated lung infection are Pseudomonas aeruginosa and Staphylococcus aureus. 6. When MSOF occurs, the most frequently involved organ and system are the kidney and respiratory system. 7. The main bacteria causing systemic invasive infection are enteric bacilli and Pseudomonas aeruginosa. Enteric bacilli infection may be enterogenic. The latter infection is chiefly the result of cross infection in hospital. CONCLUSION: The burn caused by coal mine explosion is a combined injury characterized by pathologic changes of burn as the main issue. This kind of burn has two types, i.e., gas-explosion-burn and coal-dust-explosion-burn. PMID- 10452084 TI - [The flag venous flap in the dorsal finger]. AB - OBJECTIVE: To improve the design and application of the flag flap. METHOD: Based on the anatomical characteristics of the dorsal digital veins and the venous arch, the flag flap pedicled with the veins in the dorsum of the finger was used to repair the skin defects of the ventral finger and hand in six cases. RESULTS: All flaps survived with primary healing. Three months' follow-up showed that the function and appearance of the finger were satisfactory. CONCLUSION: The design of the flap broadens its application in repairing the defects on the ventral finger and hand. PMID- 10452085 TI - [Surgical therapy of hallux valgus in the young and middle-aged female]. AB - OBJECTIVE: To correct hallux valgus in the young or middle-aged female. METHOD: Through dissecting foot specimens with hallux valgus we found that the intrinsic factor that caused hallux valgus was a "capstan" like mechanism of pressure. In order to eliminate the mechanism of the deformity, we cut off the link between the adductoris hallucis and the hallux completely. The tendon of adductoris hallucis was sutured to the fascial flap of hallucis bursa passing through a hole in the caput of the metatarsus. By this way the adductoris hallucis was fixed to the 1st metatarsus strongly, thus it pulled the 1st metatarsus directly. RESULTS: Twenty feet received this operation. 18 feet were followed up for 0.5-4 years. The rate of excellent and good results was 100%. CONCLUSION: This operation by suturing the adductoris hallucis with the fascial flap of the hallucis bursa not only can relieve pain but also restore the normal appearance of the foot. PMID- 10452086 TI - [Anatomical and clinical studies on the free posterior leg flap pedicled with the lateral popliteal cutaneous artery]. AB - OBJECTIVE: To investigate the feasibility of free transference of the posterior leg flap which is supplied by the lateral popliteal cutaneous artery. METHODS: Anatomical dissection is performed in twenty-two freshly amputated adult lower extremities. RESULTS: The discovery rate of the lateral popliteal cutaneous artery is 100%. The vessel is 20.0 +/- 5.7 cm in length with an exterior diameter of 1.28 +/- 0.25 mm at the junction of the popliteal artery. The course of the artery is constant, and it is usually accompanied by the lateral sural cutaneous nerve. Seven operations using this free flap have been performed successfully. CONCLUSION: The flap is suitable for repairing medium-sized soft tissue defects or for functional reconstruction. PMID- 10452087 TI - [Regional differences in capillary density of children's skin--an enzyme histochemical and stereological study]. AB - OBJECTIVE: The purpose of this study was to investigate the capillary distribution characteristics of the human skin. METHOD: The alkaline phosphatase enzyme histochemical and stereological methods were used to measure the length density (Lv) of capillaries of normal skin. One hundred and twenty specimens were taken from 20 different sites in 6 children cadavers. RESULT: The results showed: 1. The capillary Lv of the human skin varied with the body region. Statistically significantly (P < 0.01) greater capillary Lv was found in the head-face/neck region both in the papillary and reticular dermis than in the other parts of the body (excluding palm and sole). The smallest dermal capillary Lv was noted in the lower limb. 2. Statistically significantly (P < 0.001) higher capillary Lv was found in the papillary dermis than in the reticular dermis. CONCLUSION: The study confirms the regional and laminal differences in capillary density of the normal human skin, and it demonstrates more accurately rich blood supply in the head face/neck region, which supports the hypothesis that longer random-pattern skin flaps can be safely raised in the head-face/neck than in other parts of the body. PMID- 10452088 TI - [Microsurgical repair of the defect with large area of bare tibia]. AB - OBJECTIVE: To study the best method in repairing the defect with a large area of bare tibia. METHOD: Three hundred and twenty-two cases of large bare tibia defects were treated by three microsurgical methods. RESULTS: The bare tibiae in the 322 cases ranging from 12 x 3 cm to 24.0 x 3.5 cm were covered with myocutaneous flaps. Primary wound healing was achieved in 300 cases and delayed healing in 22 cases. All the cases were followed-up for 1 to 5 years. The function of the lower limbs was evidently improved. None of them was amputated due to deficit of skin coverage. The patients were all satisfied with the operative results. CONCLUSION: Transposition of the bridging skin flap pedicaled with the contralateral posterior tibial blood vessels can cover extensive bare tibia to avoid amputation. Other myocutaneous flaps from thoraco-umbilical, tensor muscle of fascia lata and scapular-lateral chest areas are good options too. PMID- 10452089 TI - [Advances in the study of mechanism of cicatrix contracture]. PMID- 10452090 TI - [Advance in the study on biological bases of hypertrophic scar]. PMID- 10452091 TI - Effect of tetrahydropalmatine analogs on Fos expression induced by formalin-pain. AB - AIM: To study the effect of tetrahydropalmatine (THP) analogs on Fos protein expression induced by formalin-pain and elucidate analgesic mechanism of THP analogs. METHODS: The pain response to Sprague Dawley rats was induced with formalin injected s.c. into the plantar surface of the right hindpaw. Fos protein expression in brain and spinal cord was investigated with immunohistochemistry. The numbers of Fos-like immunoreactive (FLI) neurons were counted with Leica Q570 image analyzer. RESULTS: In the groups of THP analogs and D2 antagonist spiperone, FLI neurons induced by intraperitoneal (i.p.) injection of THP analogs and spiperone were mainly located in the striatum and accumbens nucleus, and a few FLI neurons were also in sensorimotor cortex. In the D1 antagonist, D1 agonist, D2 agonist, saline and vehicle groups, FLI neurons were seldom seen in the striatum and accumbens nucleus. Moreover, the Fos protein expression induced by l-THP and spiperone could be prevented by the pre-treatment of the D2 agonist quinpirole but not D1 agonist SKF38393. In the formalin-pain group, FLI neurons were mainly distributed in ascending pain afferent system (APAS) and descending pain modulation system (DPMS). Following i.p. THP analogs, however, the numbers of FLI neurons induced by formalin-pain in the APAS, such as dorsal horn (mainly laminae I, II, IV-VI) were markedly decreased, while the numbers of FLI neurons in the DPMS, such as periaqueductal gray (PAG) and reticular paragigantocellular lateral nucleus (RPLN) were significantly increased. CONCLUSION: THP analogs enhanced the activity of brainstem DPMS by the blockade of D2 receptors in the striatum and accumbens nucleus, and sequentially inhibited the inputs of peripheral pain afferent message in spinal cord level. PMID- 10452092 TI - Effect of melatonin on production of hydroxyl radical and lactate dehydrogenase during hypoxia in rat cortical slices. AB - AIM: To study the effect of melatonin on the production of hydroxyl radical (.OH) and lactate dehydrogenase (LDH) following hypoxia in cortical slice. METHODS: Cortical slice was incubated with artificial cerebrospinal fluid (ACSF) in tube. Hypoxia was achieved by substituting 91.6% N2 and 8.4% O2. The salicylate trapping method was used to measure hydroxyl radicals generated. The content of LDH in medium after hypoxia was measured by International Federation of Clinic Chemistry (IFCC) method. RESULTS: The contents of dihydroxybenzoic acid (DHBA) were increased significantly during hypoxia and reoxygenation in cortical slice. The production of DHBA in reoxygenation was decreased concentration-dependently by melatonin, but not during hypoxia 30 min. The release of LDH during hypoxia was steadily elevated and melatonin decreased the content of LDH after hypoxia. CONCLUSION: Melatonin decreased the injury and production of .OH after hypoxia. PMID- 10452093 TI - Inhibitory effects of melatonin on free intracellular calcium in mouse brain cells. AB - AIM: To study the effects of melatonin (Mel) on cortical intrasynaptosomal calcium concentration in old mice and on [Ca2+]i elevation induced by Bay-K-8644, KCl, and sodium l-glutamate in isolated brain cells of neonatal mouse, and to determine the antiaging mechanism of Mel. METHODS: [Ca2+]i was measured in an RF 5000 recording spectrofluorophotometer by preloading the synaptosomes or cells with Fura 2-AM. RESULTS: Long term of administrating Mel inhibited the overload of [Ca2+]i in old mouse cerebral cortex. The [Ca2+]i in both high (20 mg.L-1) and low dose (1 mg.L-1) of Mel groups was reduced from (434 +/- 32) nmol. L-1 (the older control group) to (330 +/- 41) and (313 +/- 56) nmol.L-1, respectively, P < 0.01. Mel 0.01, 0.1, 1, and 3 mumol.L-1 remarkably reduced [Ca2+]i elevations in isolated newborn mouse brain cells induced by Bay-K-8644, KCl, and Glu. CONCLUSIONS: The inhibitory effect of Mel on neuronal [Ca2+]i overload is involved in its antiaging effect. PMID- 10452094 TI - Attenuation of scopolamine-induced deficits in navigational memory performance in rats by bis(7)-tacrine, a novel dimeric AChE inhibitor. AB - AIM: To study the effects of 1,7-N-heptylene-bis-9,9'-amino-1,2,3,4 tetrahydroacridine [bis(7)-tacrine], a novel dimeric acetylcholine-sterase inhibitor (AChEI) derived from 9-amino-1,2,3,4-tetrahydroaminoacridine (tacrine), on scopolamine-induced spatial memory impairment. METHODS: The effects of bis(7) tacrine were investigated on the 5-d performance of young adult rats in the Morris water maze. The latency to find the platform in the water maze was measured to evaluate performance. Tacrine was used as a reference drug. RESULTS: Scopolamine (0.3 mg.kg-1, i.p.) resulted in an increase in latency period (> 100% increase) as compared with saline treated controls. Both bis(7)-tacrine and tacrine lessened the increased latency induced by scopolamine to the level of saline control group. The relative potency of bis(7)-tacrine (0.35 mumol.kg-1, i.g. or i.p.) to shorten the escape latency was 24 or 12 times of tacrine (8.52 mumol.kg-1 i.g., 4.26 mumol.kg-1 i.p.) following i.g. or i.p. administration, respectively. There appeared to be an inverse bell-shape dose-dependent effect for both compounds tested. CONCLUSION: Bis(7)-tacrine is a more potent and orally active AChEI than tacrine, and has potential for the palliative treatment of Alzheimer disease. PMID- 10452095 TI - Population distributions of allele frequency of apolipoprotein E by age and gender in Han Chinese. AB - AIM: To study apolipoproteins E (ApoE) allele frequency in Han Chinese based on age and gender from Shanghai metropolitan area. METHODS: Healthy Han Chinese people (F: 237 and M: 412) were involved in this study. ApoE gene was amplified by PCR using the forward primer: 5'-GGC ACG GCT GTC CAA GGA GCT-3' and reverse primer: 5'-GAT GGC GCT GAG GCC GCG CT-3'. The PCR product was digested directly with 5 units of CfoI and separated by a 20% polyacrylamide nondenaturing gel. RESULTS: ApoE*3 was the commonest allele which accounted for 86.4% of the isoforms, and ApoE*2 and ApoE*4 accounted for 6.2% and 7.5%, respectively. The allele and genotype frequencies were in Hardy-Weinberg equilibrium by comparison with that of the corresponding theoretical distribution (P > 0.05). CONCLUSION: The frequencies of ApoE*2, ApoE*3, and ApoE*4 were demonstrated in the normal Chinese population. PMID- 10452096 TI - Efflux transport of [3H]GABA across blood-brain barrier after cerebral ischemia reperfusion in rats. AB - AIM: To study whether the efflux transport of [3H]GABA across the blood-brain barrier (BBB) would be enhanced after cerebral ischemia-reperfusion. METHODS: Brain efflux index (BEI) of [3H]GABA was determined in ischemic-reperfused rats after [3H]GABA or [3H]GABA combined with unlabeled GABA or probenecid (Pro) was microinjected into the parietal cortex area 2 (Par 2), and brain uptake of Evans blue (EB) was assessed after i.v. EB. RESULTS: BEI in rats subjected to 10-min ischemia and 30-min, 2-h, 6-h, or 24-h reperfusion were 67%, 83%, 92%, and 87%, respectively, which were higher than that in control (58%). The brain uptake of EB was also considerably increased. Unlabeled GABA or Pro obviously decreased BEI in normal or 6-h reperfused rats, but GABA had no obvious effect on that in 5-min reperfused rats. CONCLUSION: The efflux transport of [3H]GABA was markedly enhanced after cerebral ischemia-reperfusion in rats. PMID- 10452097 TI - Effect of basic fibroblast growth factor on focal ischemic injury and antioxidant enzyme activities. AB - AIM: To explore the effect of basic fibroblast growth factor (bFGF) on focal cerebral ischemic injury and antioxidant enzyme activities. METHODS: Rats underwent 24-h middle cerebral artery occlusion by intraluminal suture. Infarction volume was shown with staining and quantitated by image analysis system. Neurologic deficit scores were determined with a 0-5 grade scale. Antioxidant enzyme activities of forebrains were detected. RESULTS: bFGF (45 micrograms.kg-1.h-1 i.v. for 3 h, started 5 min after the onset of ischemia) showed potent neuroprotective effects. Infarction volumes were decreased from 272 mm3 +/- 22 mm3 (saline-treated) to 201 mm3 +/- 30 mm3 (bFGF-treated). Neurologic deficit scores were decreased from 3.6 +/- 1.5 (saline-treated) to 2.3 +/- 1.6 (bFGF-treated). Focal cerebral ischemia induced an increase in the activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), but a decrease in the level of glutathione s-transferase (GSH-ST). Treatment of bFGF further increased the CAT activity but had no effect on the activities of SOD, GSH-PX, and GSH-ST. CONCLUSION: bFGF has a neuroprotective effect against focal cerebral ischemic injury. The elevation of CAT activity by bFGF may be involved in this effect. PMID- 10452098 TI - Effects of agmatine on tolerance to and substance dependence on morphine in mice. AB - AIM: To study the effects of agmatine on tolerance to and dependence on morphine. METHODS: Inhibitory effects of agmatine on tolerance to and substance dependence on morphine were observed in mouse tolerant models and in mouse jumping test, respectively. RESULTS: Agmatine 0.125-2.5 mg.kg-1 prevented the development of tolerant to morphine in a dose-dependent manner. Pretreatment of mice with morphine induced an over 3-fold increase in analgesic ED50 (20.1, 14.4-28.0 mg.kg 1) than those with normal saline (6.3, 5.1-7.8 mg.kg-1). Pretreatment of mice with both of agmatine and morphine made morphine loss the ability to induce tolerance. Withdrawal jumps and loss in body weight induced by naloxone in morphine-dependent mice were prevented by agmatine (2.5-10 mg.kg-1) in a dose dependent manner. ED50 of naloxone (21.4, 18.4-24 mg.kg-1) required to precipitate withdrawal jumps in mice pretreated with both agmatine and morphine was 8 times higher than that with morphine alone (2.5, 2.1-2.8 mg.kg-1). These effects of agmatine were blocked by idazoxan. CONCLUSION: Agmatine prevented tolerance to and substance dependence on morphine in mice by activation of imidazoline receptors. PMID- 10452099 TI - Alpha-momorcharin inhibits HIV-1 replication in acutely but not chronically infected T-lymphocytes. AB - AIM: To identify the anti-human immunodeficiency virus type 1 (HIV-1) activities of alpha-momorcharin (alpha-MMC) from Momordica charantia in acutely and chronically infected T-lymphocytes. METHODS: The anti-HIV activities of alpha-MMC were examined by 1) the inhibition of syncytia formation induced by HIV-1 III B; 2) reduction of p24 core antigen expression level and decrease in numbers of HIV antigen positive cells in acutely and chronically infected cultures. The cytotoxic effects of alpha-MMC was tested by trypan blue dye exclusion or colorimetric MTT assay. RESULTS: alpha-MMC was found to obviously inhibit HIV-1 III B-inducing C8166 syncytia formation and markedly reduced both expression of p24 core antigen and the numbers of HIV antigen positive cells in acutely but not chronically HIV-1-infected culture. The median effective concentration (EC50) in these assays were 0.016, 0.07, and 0.32 mg.L-1, respectively. CONCLUSION: alpha MMC is a unique component of momorcharin with anti-HIV activity, and markedly inhibited HIV-1 replication in acutely but not chronically HIV-1-infected T lymphocytes. PMID- 10452100 TI - Pharmacokinetics and peripheral platelet counts after a single dose of recombinant human thrombopoietin in rhesus monkeys. AB - AIM: To study the pharmacokinetics and the change of peripheral platelet counts after a single dose of recombinant human thrombopoietin (rhTpo). METHODS: After i.v. or s.c. injections of rhTpo in 12 rhesus monkeys, rhTpo concentration in serum was determined by ELISA. Platelets were counted by automatic microcell counter. RESULTS: The terminal half-lives of rhTpo were 12-18 h. AUC following s.c. were linearly increased with dose, while Cls were 0.061, 0.08, and 0.07 L.kg 1.h-1 in s.c. 0.5, 2, and 8 micrograms.kg-1 groups, respectively. Bioavailability was 0.50 +/- 0.18 after s.c. Single dose of rhTpo was associated with an increase in platelets (55.9%-107.4%, P < 0.05) in a dose-related manner. The peak response and the sustained days of platelet increase were dose-related. The degree of platelet increase (% x time) correlated with the systemic exposure to rhTpo (C x time). CONCLUSION: rhTpo behaved as a linear pharmacokinetics in the monkey within dose range of 0.5-8 micrograms.kg-1. PMID- 10452101 TI - Inhibitory actions of polysaccharide sulfate on action potentials and contraction of papillary muscles in guinea pigs. AB - AIM: To examine the effects of polysaccharide sulfate (PSS) on the action potentials and contractile force in guinea pig papillary muscles. METHODS: Using intracellular microelectrode to record fast (FAP) and slow (SAP) action potentials. RESULTS: PSS (> or = 50 mg.L-1) caused concentration-dependent decreases in the contractile force and the action potential duration (APD) of FAP without affecting the resting potential (RP), action potential amplitude (APA), and maximal upstroke velocity (Vmax). The Vmax, APA, and APD of BaCl2-induced SAP were concentration-dependently decreased by PSS (> or = 15 mg.L-1) and the effects were antagonized by isoprenaline (1 mumol.L-1). The APA and APD of isoprenaline-induced SAP were decreased by PSS (> or = 15 mg.L-1) in a concentration-dependent manner and the effects were attenuated by elevation of extracellular Ca2+ concentration. CONCLUSION: PSS selectively inhibited the slow inward current. PMID- 10452102 TI - Dual effects of tetrandrine on calcium-activated potassium channels in pulmonary artery smooth muscle cells. AB - AIM: To study the effects of tetrandrine (Tet) on calcium-activated potassium channels. METHODS: Inside-out configuration of patch-clamp single channel recording techniques. RESULTS: Tet 7.5 and 15 mumol.L-1 increased the open probability from control value 0.251 +/- 0.012 to 0.340 +/- 0.013 and 0.415 +/- 0.011, respectively (P < 0.01), decreased the close time from (61 +/- 15) ms to (33 +/- 10) and (28 +/- 11) ms, respectively (P < 0.01). But Tet 30 mumol.L-1 decreased the open probability and open time to (0.114 +/- 0.008) and (1.47 +/- 0.09) ms, respectively [P < 0.01 vs control (0.251 +/- 0.012) and (20 +/- 8) ms]. CONCLUSION: Tet has concentration-dependent dual effects on KCa channels in isolated rat pulmonary smooth muscle cells. PMID- 10452103 TI - Relationship between adenosine-induced vascular effects and ATP-sensitive K+ channels. AB - AIM: To study the relationship between adenosine (Ade) receptors and adenosine 5' triphosphate (ATP)-sensitive potassium (KATP) channels in rat aorta. METHODS: Isolated rat aorta rings were suspended for isometric force recording. The vascular effects of Ade were assessed in the presence or absence of functional endothelium. The interactions of Ade and pinacidil (Pin) or glibenclamide (Gli) were investigated. RESULTS: In isolated aorta preconstricted with KCl 20 mmol.L 1, Ade 3-300 mumol.L-1 induced relaxation in a concentration-dependent manner; and in 48/99 preparations from 32 rats, Ade induced initial transient constriction followed by sustained relaxation. When the functions of KATP channels were blocked with Gli 1 or 100 mumol.L-1, effects of Ade were characterized by vasoconstriction rather than vasorelaxation. The combination of Pin 1 mumol.L-1 with Ade 100 mumol.L-1 showed no synergic vasodilatory effects and did not affect Ade-induced vasoconstriction. After the removal of endothelium, Ade still induced vasoconstriction and vasorelaxation, and the constrictive effects showed no difference from those in the presence of endothelium, but the potency of vasodilatory effects became weaker with slower decrease in tension. CONCLUSION: The activation of KATP channels is involved in Ade receptor-induced vasodilation. PMID- 10452104 TI - Dinoprostone potentiates cytokines and lipopolysaccharides inducing nitric oxide production in cultured rat hepatocytes. AB - AIM: To study the effects of dinoprostone (Din) on nitric oxide (NO) production induced by lipopolysaccharides (LPS) and cytokines mixture (CM) in cultured rat hepatocytes, and the role of cAMP signaling pathway in the induction process. METHODS: Rat hepatocytes were incubated with indometacin (Ind), Din, forskolin (For), or dibutyryl cyclic GMP (db cGMP) in a medium containing CM (LPS plus TNF alpha, IL-1 beta, and IFN-gamma) for 24 h. NO production in the cultured supernatant was measured with the Griess reaction. Intracellular cAMP level was measured with radioimmunoassay. RESULTS: NO production stimulated by CM was markedly attenuated by Ind in the cultured supernatant, whereas Din and For increased nitrite concentration in the medium. In the presence of CM, both Din and For increased intracellular cAMP level, and had no effect on cAMP level in absence of the stimuli; db cGMP had no influence on the cAMP and NO production both in the presence and absence of same stimuli. CONCLUSION: Din potentiates cytokines and LPS to induce NO production in rat hepatocytes, and the induction may be regulated via intracellular second messenger cAMP signaling pathway. PMID- 10452105 TI - Securinine induced apoptosis in human leukemia HL-60 cells. AB - AIM: To study whether securinine might induce apoptosis in human leukemia HL-60 cells. METHODS: Inhibition of proliferation was measured using MTT assay. The amount of apoptotic cells was measured by flow cytometry. DNA fragmentation was visualized by DNA agarose gel electrophoresis and the cellular changes were observed by electron microscope. RESULTS: Securinine 5-80 mg.L-1 elicited typical apoptosis morphological changes and DNA fragmentation in a concentration dependent manner in HL-60 cells. Securinine inhibited HL-60 cell proliferation in a time- and concentration-dependent manner. The IC50 and 95% confidence limits were 27 (15-47) mg.L-1 after 12-h treatment with securinine. CONCLUSION: Securinine induced apoptosis in HL-60 cells. PMID- 10452106 TI - Inhibitory effects of nitric oxide and interleukin-10 on production of tumor necrosis factor alpha, interleukin-1 beta, and interleukin-6 in mouse alveolar macrophages. AB - AIM: To observe the effects of nitric oxide and interleukin-10 (IL-10) on inflammatory reaction in mouse alveolar macrophages (AM). METHODS: AM from mice were stimulated by lipopolysaccharides (LPS) 10 mg.L-1 and nitric-oxide synthase inhibitor, S-methylisothiorea sulfate (SMT) or nitric-oxide donor, S-nitroso-N acetyl-D, L-penicillamine (SNAP). The production of tumor necrosis factor alpha (TNF alpha), IL-1 beta, IL-6, and IL-10 by AM were measured by ELISA. RESULTS: After LPS-stimulation, TNF alpha, IL-1 beta, and IL-6 peaked at 6, 12, and 24 h, respectively by AM. SMT inhibited LPS-induced nitric oxide release and increased IL-1 beta and IL-6 secretions in AM, but the TNF alpha levels remained unchanged. SNAP had inhibitory effects on IL-1 beta and IL-6 secretions in a concentration dependent manner, but exerted no effect on TNF alpha release. TNF alpha, IL-1 beta, and IL-6 secretions were inhibited by recombinant IL-10, but the cytokines release was upregulated by anti-IL-10 monoclonal antibody. CONCLUSION: Both endogenous and exogenous nitric oxide and IL-10 had inhibitory effects on the LPS induced TNF alpha, IL-1 beta, and IL-6 secretions in mouse AM. PMID- 10452107 TI - Basic fibroblast growth factor enhanced LAK cell cytotoxicities against human bladder neoplasm cells. AB - AIM: To study the effects of basic fibroblast growth factor (bFGF) on the proliferation of lymphokine-activated killer (LAK) cells from patients with bladder cancer and LAK cells cytolysis against bladder tumor cells. METHODS: LAK cell proliferation was assayed in the presence of various concentrations of bFGF combined with interleukin-2 (IL-2) by cell count. Cytotoxicity of LAK cells against bladder cancer cell line EJ cells and bladder tumor cells (BTC) from patients was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. RESULTS: The proliferation of peripheral blood monocytes (PBMC) was inhibited by bFGF 5 micrograms.L-1. bFGF did not affect the stimulation of LAK cells induced by IL-2. The LAK cell numbers in the combination of IL-2 with bFGF were not significantly different compared with that treated with IL-2 alone. bFGF enhanced cytotoxicity of LAK cells against bladder cancer cell line EJ cells or BTC, respectively. CONCLUSION: Although the proliferation of PBMC was inhibited by bFGF, bFGF increased LAK cell cytotoxicity against bladder neoplasm cells. PMID- 10452108 TI - Velvet antler polypeptides promoted proliferation of chondrocytes and osteoblast precursors and fracture healing. AB - AIM: To study the effects of velvet antler (VA) total polypeptides (VATP) and VA polypeptides, VAP-A, VAP-B, and VAP-C on proliferation of chondrocytes and osteoblast precusors. METHODS: Chondrocytes (rabbit and human fetus) and osteoblast precusors (chick embryo) were incubated in the culture medium containing VATP or VAP-A, VAP-B, and VAP-C. [3H]TdR incorporation into DNA was measured. Fracture healing-promoting action of VATP was determined in rats. RESULTS: VATP 50-200 mg.L-1 and VAP-B 12.5, 25, and 50 mg.L-1 showed most marked proliferation-promoting activity for rabbit costed chondrocytes and increased incorporation of [3H]TdR from (73 +/- 9) Bq (control group) to (272 +/- 55), (327 +/- 38), and (415 +/- 32) Bq, respectively (P < 0.01). The activity of VAP-A was weaker than that of VAP-B, and VAP-C had no activity. VATP 10 and 20 mg.kg-1 by local injection into the cross-section fracture area accelerated healing of radial fracture. The healing rate of VATP-treated group was higher (75%) than that of control group (25%) (P < 0.05). CONCLUSION: VATP accelerated fracture healing by stimulating proliferation of chondrocytes and osteoblast precursors. PMID- 10452109 TI - Syntheses and biological activities of chiral piperidines-tachykinin NK3 antagonists. AB - AIM: To develop nonpeptide tachykinin NK3 antagonists. METHODS: Five tachykinin NK3 antagonists were synthesized. Receptor binding assay and oral absorption study were made. RESULTS: The 4,4-disubstituted piperidine compounds (1b, 1c, and 1d) showed stronger activities (IC50 = 5.9, 6.2, and 11 nmol.L-1, respectively) than the monosubstituted ring compound 1e (IC50 = 17 nmol.L-1). 4-Phenyl (1b) and 4-phenylsulfonylmethyl (1c) compounds were more active than the 4-fluorobenzyl compound (1d). All antagonists were found to be orally absorbable, the T1/2 of 1b (6.4 h) was more than three-fold longer than that of 1a (1.9 h). CONCLUSION: Compound 1b had the best binding activity (IC50 = 5.9 nmol.L-1) and the best AUC (2081 micrograms.h.L-1). PMID- 10452110 TI - Protirelin (thyrotropin-releasing hormone) in thyroid gland: possible involvement in regulation of thyroid status. AB - AIM: To establish the presence of the hypothalamic hormone protirelin (thyrotropin-releasing hormone, TRH) in human thyroid and to investigate whether the concentration of this peptide in the thyroid gland is sensitive to thyroid status. METHODS: A procedure has been developed for the determination of TRH in the thyroid gland, distinct from TRH-like peptides which also react with TRH antibody. RESULTS: Human thyroid was shown to contain both authentic TRH and TRH like peptides, a similar pattern was seen in a range of animal thyroids. The concentrations of TRH in non-active goiter thyroids were substantial (41.6-248 pmol.g-1); in contrast the thyroids from hyperthyroid patients contained very little TRH (0.01-2.52 pmol.g-1). CONCLUSION: The physiologic role of TRH in the thyroid is not known but the large difference between the concentrations of this hormone in non-active and hyperactive thyroids suggests that thyroidal TRH may be involved in the regulation of thyroid status. PMID- 10452111 TI - Antagonistic effects of berbamine on [Ca2+]i mobilization by KCl, norepinephrine, and caffeine in newborn rat cardiomyocytes. AB - AIM: To study the effects of berbamine (Ber) on intracellular calcium concentration ([Ca2+]i) mobilized by KCl depolarization, norepinephrine (NE), and caffeine. METHODS: [Ca2+]i was measured with fluorescent intensity (FI) by confocal microscope in single cultured cardiomyocytes of newborn rats loaded with Fluo 3-AM 2 mumol.L-1. RESULTS: FI value of [Ca2+]i in control level was 248 +/- 70 in the presence of extracellular calcium 1.5 mmol.L-1 and was not changed by Ber 3-30 mumol.L-1. KCl (60 mmol.L-1)- and NE (30 mumol.L-1)-induced [Ca2+]i mobilizations were inhibited (P < 0.01) by Ber 30 mumol.L-1, similar to that of verapamil (Ver). The inhibitory effect of Ber on [Ca2+]i induced by KCl was further increased (P < 0.05) in the presence of egtazic acid 3 mmol.L-1, but that on [Ca2+]i induced by NE was not changed. The [Ca2+]i mobilized by caffeine 80 and 160 mumol.L-1 in D-Hanks' solution was not affected (P > 0.05) by Ber and Ver. CONCLUSION: Ber possessed the antagonistic effects on [Ca2+]i increases via voltage-dependent Ca2+ channel and receptor-operated Ca2+ channel in newborn rat cardiomyocytes, but without effect on intracellular Ca2+ release. PMID- 10452112 TI - In vitro cytotoxicity of salvicine, a novel diterpenoid quinone. AB - AIM: To study the in vitro cytotoxicity of 4,5-seco-5,10-friedo-abieta-3,4 dihydroxy-5(10),6,8,13-tetraene-11, 12-dione (salvicine), a novel diterpenoid quinone compound on human tumor cell lines and its effect on cell cycle progression. METHODS: Growth inhibition of human tumor cells was measured by microculture tetrazolium assay (MTT). Cell cycle was analyzed by flow cytometry. RESULTS: Exposing tumor cell lines tested to salvicine for 72 h, in comparison with reference drugs vincristine (VCR) and etoposide (VP-16), salvicine was as cytotoxic as VP-16 and weaker than VCR in 3 leukemia cell lines. For 12 solid tumor cell lines, salvicine exhibited cytotoxic activities and was over 5.41- and 4.15-fold stronger than VCR and VP-16, respectively. Salvicine presented better activities especially against gastric and lung carcinoma cell lines. Exposing K562 leukemia cells to 9 graded concentrations of salvicine (from 0.39 to 100 mumol.L-1) for 24 h and to salvicine 10 mumol.L-1 for 7 different periods (from 1 to 48 h), the growth inhibition of cells was enhanced along with increased concentration or prolonged exposure. Cell cycle analysis demonstrated that salvicine arrested K562 cells in G1 phase and this effect was also heightened with increased concentration or extended exposure. CONCLUSION: Salvicine exhibited potent cytotoxic activities against various human tumor cell lines, and blocked K562 leukemia cells in G1 phase of cell cycle. PMID- 10452113 TI - Characterization of outward potassium current in embryonic chick heart cells. AB - AIM: To characterize a voltage-dependent outward K+ current in cultured heart cells of 14-16-day-old embryos of yellow chick. METHODS: The patchclamp technique in the whole-cell configuration was used. RESULTS: The kinetics and the pharmacology of the outward K+ current in our cell mold were different from those described in white chick. Like the calcium-activated K+ current, blocker of calcium channel, CdCl2, eliminated the current of more than 95%. Isoproterenol provoked an increase of peak amplitude (137% +/- 47%, n = 16 cells) and acceleration of activation kinetics in the outward K+ current (the time reaching a peak current reduced from 36 ms +/- 10 ms to 16 ms +/- 9 ms). This effect of isoproterenol was mimiced by cAMP. In addition, a frequency-dependent decrease in peak amplitude of the current occurred after cAMP-induced phosphorylation. CONCLUSION: There are species- and/or cell-type-specific difference in the K+ channels properties. In embryonic yellow chick heart cells, the phospholation of channel could not only modulate the activation kinetic properties of the calcium activated potassium channel, but also change their recovery kinetics. PMID- 10452114 TI - Involvement of NMDA and non-NMDA receptors in transmission of spinal visceral nociception in cat. AB - AIM: To study the role of N-methyl-D-aspartic acid (NMDA) and non-NMDA receptors in processing nociceptive visceral information in the spinal cord. METHODS: The firing of spinal dorsal horn neurons to colorectal distension (3-15 kPa, 20 s) by inflation with air of latex balloon was recorded in 25 anesthetized cats. RESULTS: 1) According to the patterns of responses to colorectal distension, the neurons with increase and decrease in firing were classified as excitatory and inhibitory, respectively. The former consisted of 17 short-latency abrupt (SLA) neurons, 11 short-latency sustained (SLS) neurons, 9 long-latency (LL) neurons. The 15 inhibited (Inh) neurons were recorded. 2) Microelectrophoretic administration of NMDA, quisqualic acid (QA), and kainic acid (KA) activated 67.6%, 78.4%, and 59.5% of the colorectal distension-excited neurons tested. Also, 60%, 86.7%, and 53.3% of Inh neurons were activated by these 3 amino acids. 3) Colorectal distension-induced excitatory responses were reduced by 35% +/- 10% and 65% +/- 14% by a selective NMDA receptor antagonist d,l-2-amino-5 phosphonovalerate (APV) and a selective non-NMDA receptor antagonist 6,7-dinitro quinoxaline-2,3-dione (DNQX), respectively. Such DNQX-induced inhibition was significantly more potent than that by APV (P < 0.05). Colorectal distension induced inhibitory responses were partially relieved by 30%-50% in 3/7 Inh neurons by DNQX, but not APV. CONCLUSION: Both NMDA and non-NMDA receptors are involved in transmission and/or modulation of spinal visceral nociceptive information and non-NMDA receptors may play more important role than NMDA receptors. PMID- 10452115 TI - Intraventricular vascular endothelial growth factor antibody increases infarct volume following transient cerebral ischemia. AB - AIM: To clarify the role of vascular endothelial growth factor (VEGF) in neuronal damage induced by cerebral ischemia. METHODS: Expression of VEGF in adult rat brain was measured by immunohistochemistry. Transient middle cerebral artery occlusion (MCAO) model was induced by placing a nylon thread in the lumen of the internal carotid artery. The infarct volume was shown with 2,3,5 triphenyltetrazolium chloride (TTC) staining and quantitated by computer image analyzer with and without VEGF antibody treatment. RESULTS: VEGF expression was widely distributed in neuronal cells besides vascular endothelial cells, and the neuronal distribution of VEGF was specific. After intraventricular treatment with VEGF antibody (0.1 g.L-1 daily, for 7 d following the ischemia), infarct volume in the antibody treatment was increased versus vehicle-treated rats [(21.6 +/- 2.7 vs 16 +/- 6) mm3, P < 0.05] respectively. CONCLUSION: Intraventricular injection of VEGF antibody increased the infarct volume after focal cerebral ischemia in rats, suggesting that expression of neuronal VEGF may be one of neuronal protective mechanisms. PMID- 10452116 TI - Effects of 7-nitroindazole on long-term potentiation induced by l-clausenamide and high-frequency stimulation in rat hippocampus in vivo. AB - AIM: To study the antagonistic effect of selective neuronal nitric-oxide synthase (nNOS) inhibitor 7-nitroindazole on the long-term potentiation (LTP) induced by l clausenamide (Cla) in rat hippocampus in vivo. METHODS: Population spike (PS) of evoked potentials was determined by extracellular recording technique in the hippocampal dentate gyrus (DG) of anesthetized rats. RESULTS: 7-Nitroindazole 2 nmol icv blocked the induction of LTP elicited by high-frequency (100 Hz) stimulation or Cla 5 nmol icv (P < 0.01), and L-arginine 225 mg.kg-1 i.p. prevented the action of 7-nitroindazole (P < 0.01). CONCLUSION: Nitric oxide produced by nNOS plays a role in the induction of Cla-induced LTP in hippocampus. PMID- 10452117 TI - Inhibition by baclofen of NMDA-activated current in rat dorsal root ganglion neurons. AB - AIM: To explore the modulatory effect of baclofen on NMDA-activated current in rat dorsal root ganglion (DRG) neurons. METHODS: Whole-cell patch-clamp technique was used to record NMDA-activated current in isolated DRG neurons. Drugs were applied by rapid solution exchange. RESULTS: Preapplication of baclofen 1-100 mumol.L-1 induced a concentration-dependent inhibition of the inward NMDA activated current markedly. NMDA (100 mumol.L-1)-activated current was inhibited by 52% +/- 14% (n = 11, P < 0.01) by preapplication of baclofen 100 mumol.L-1. The inhibitory effect of baclofen was reversible, and was removed by saclofen 100 mumol.L-1, which was a selective antagonist of GABAB receptor. CONCLUSION: Preapplication of baclofen exerts an inhibitory effect on NMDA-activated current in the primary sensory neurons. PMID- 10452118 TI - Effects of excitatory amino acids and nimodipine on calcium currents in cultured rat cortical neurons. AB - AIM: To study the effect of excitatory amino acid (EAA) and calcium channel blocker on neuronal calcium channels. METHODS: With path-clamp technique (whole cell recording), the effects of Bay-K-8644, cesium glutamate, potassium aspartate, and nimodipine (Nim) on calcium currents (ICa) in cultured cortical neurons of neonatal rats were studied. RESULTS: ICa was raised obviously by Bay-K 8644 and glutamate. ICa was raised concentration-dependently by aspartate (0.5, 5, 50 mmol.L-1), with increasing rates 15% +/- 3%, 37% +/- 3%, and 53% +/- 6%, respectively. The inhibition of ICa was obvious while adding Nim in the bath solution. With Nim 10 mumol.L-1, the inhibitory rate was 46% +/- 4%. CONCLUSION: EAA had increasing effects on neuronal calcium currents and Nim inhibited Ca2+ influx in neurons. PMID- 10452119 TI - Nitric oxide derived from endothelial cells inhibits Na+/H+ exchange in rabbit platelets activated by thrombin. AB - AIM: To study the effect of nitric oxide (NO) derived from endothelial cells on Na+/H+ exchange in rabbit platelets activated by thrombin. METHODS: Intracellular Ca2+ ([Ca2+]i) and intracellular pH (pHi) were measured by the dual-wavelength fluorophotometer with the fluorescent probes Fura-2 and 2',7'-biscarboxyethyl-5,6 carboxyfluorescein (BCECF). Effects of NO on rabbit platelets were tested by cultured bovine endothelial cells (BAEC). RESULTS: BAEC (0.1-1 x 10(9).L-1) inhibited thrombin (100 U.L-1)-induced platelet aggregation in a concentration dependent manner. This inhibiting effect was abolished by preincubating BAEC with NG-nitro-L-arginine 1 mmol.L-1. When the [Ca2+]i store was depleted with ionomycin in the presence of egtazic acid (EGTA), the increase in pHi induced by thrombin was inhibited. Refilling intracellular Ca2+ store partially reversed this effect. BAEC 2 x 10(8).L-1 inhibited thrombin (100 U.L-1)-induced elevation of pHi and mobilization of intracellular Ca2+ store (P < 0.01). No direct effect of endothelial cells on unstimulated rabbit platelets was observed. CONCLUSION: NO derived from endothelial cells inhibited thrombin-induced rabbit platelet activation by inhibiting thrombin-induced [Ca2+]i mobilization and then inhibiting the consequent Na+/H+ exchange in rabbit platelets. PMID- 10452120 TI - Effect of protopine on cytosolic Ca2+ in rabbit platelets. AB - AIM: To study the influence of protopine (Pro) on the cytoplasmic free Ca2+ concentration ([Ca2+]i) in rabbit platelets. METHODS: Measurement of [Ca2+]i of platelets in vitro by Fura 2-AM fluorescence technique. RESULTS: In the presence of CaCl2 1 mmol.L-1, Pro 10, 20, and 40 mumol.L-1 attenuated the rise in [Ca2+]i evoked by ADP from (420 +/- 57) to (320 +/- 26), (264 +/- 21), and (180 +/- 14) nmol.L-1, respectively, by arachidonic acid (AA) from (280 +/- 36) to (210 +/- 17), (184 +/- 21), and (143 +/- 16) nmol.L-1, respectively, and by platelet activating factor (PAF) from (350 +/- 42) to (282 +/- 31), (223 +/- 30), and (165 +/- 15) nmol.L-1, respectively. In the presence of egtazic acid 1 mmol.L-1, Pro 10, 20, and 40 mumol.L-1 reduced the Ca2+ release induced by ADP, AA, and PAF, respectively. Pro 10, 20, and 40 mumol.L-1 also decreased ADP-, AA-, and PAF induced Ca2+ influx. CONCLUSION: Pro inhibited not only Ca2+ release but also the influx of Ca2+. PMID- 10452121 TI - Ischemic preconditioning mediated by activation of KATP channels in rat small intestine. AB - AIM: To study whether the protective effects of ischemic preconditioning against rat small intestine ischemia/reperfusion injury could be mediated by KATP channel opener. METHODS: Preconditioning (Pc) was induced by 3 cycles of 8-min superior mesenteric artery (SMA) occlusion and 10-min reperfusion before prolonged ischemia. Cromakalim (Cro 75 micrograms.kg-1) and glibenclamide (Gli 8 mg.kg-1) were injected i.v. 10 min before prolonged ischemia and Pc, respectively. RESULTS: Compared with ischemic reperfusion (IR) group, Pc before prolonged ischemia (Pc + IR) decreased LDH release [(380 +/- 55) vs (559 +/- 49) U.L-1, P < 0.05], attenuated intestinal edema [wet weight/dry weight (WW/DW), 5.6 +/- 0.6 vs 6.34 +/- 0.29, P < 0.05], ameliorated intestinal histological damage (grading scale, 3.4 vs 5.7, P < 0.01), and improved reperfusion-induced hypotension. These effects of Pc were mimicked by Cro [LDH, (298 +/- 40) vs (559 +/- 49) U.L-1, P < 0.05; WW/DW, 5.6 +/- 0.4 vs 6.34 +/- 0.29, P < 0.05; grading scale, 3.6 vs 5.7, P < 0.01] and abolished in the presence of Gli [LDH, (624 +/- 44) vs (559 +/- 49) U.L-1; WW/DW, 6.6 +/- 0.6 vs 6.34 +/- 0.29; grading scale, 5.7 vs 5.7; P > 0.05] compared with IR group, respectively. CONCLUSION: Ischemic preconditioning on the rat small intestine is mediated by activation of KATP channels. PMID- 10452122 TI - Effects of simvastatin on left ventricular hypertrophy and function in rats with aortic stenosis. AB - AIM: To investigate the effects of simvastatin (Sim) on left ventricular hypertrophy in rats with pressure-overload cardiac hypertrophy. METHODS: The left ventricular hypertrophy (LVH) of rats was induced by partly occluding abdominal aorta below right renal artery. The rats were given i.g. Sim (1.8 and 3.6 mg.kg 1.d-1) for 8 wk. Three days after operation, left ventricular function was measured. Then the left ventricle (LV) + septum and the right ventricle (RV) were weighed. Hydroxyproline content of LV was measured. RESULTS: Eight weeks later, in the LVH group, LV weight (LVW), LVW/body weight (BW), LVW/RV weight (RVW), LV ending diastolic pressure (LVEDP), and hydroxyproline content increased by 36%, 51%, 28%, 92%, and 23%, respectively (all P < 0.01) compared with the sham group. LV + dp/dtmax and -dp/dtmax decreased by 39.2% and 39.4% (all P < 0.01). After the rats were given i.g. Sim 3.6 mg.kg-1.d-1, LVW, LVW/BW, LVW/RVW, left ventricle ending diastolic pressure (LVEDP), and hydroxyproline content decreased by 22%, 21%, 23%, 24%, and 11% compared with LVH group (all P < 0.01), LV + dp/dtmax and -dp/dtmax increased by 60% and 32% (all P < 0.01). CONCLUSION: Sim inhibited development of LV hypertrophy and improved LV function in rats with aortic stenosis. PMID- 10452123 TI - Inhibitory effects of captopril on hypoxia-induced proliferation and collagen synthesis in pulmonary vascular smooth muscle cells. AB - AIM: To study the effect of captopril (Cap) on hypoxia-induced proliferation and collagen synthesis in vascular smooth muscle cells (VSMC). METHODS: VSMC were isolated from rabbit pulmonary artery. Cultured VSMC were evaluated by incorporation of [3H]thymidine and [3H]proline, cell number, and intracellular calcium concentration ([Ca2+]i). RESULTS: Pretreatment of pulmonary VSMC with Cap 1 mumol.L-1 blocked hypoxia-induced increase in cell number and incorporation of [3H]proline and [3H]thymidine, which were decreased 25%, 21%, and 36%, respectively, as compared with hypoxic control. It also inhibited the increase of intracellular Ca2+ concentration under hypoxic condition. Addition of nifedipine inhibited hypoxia-stimulated increase in the collagen, DNA synthesis, and [Ca2+]i. Bay-K-8644 increased cell number (35%), DNA (55%), collagen synthesis (36%), and [Ca2+]i (33%) in pulmonary VSMC, that was completely abolished by Cap 1 mumol.L-1. CONCLUSION: Cap inhibited hypoxia-induced proliferation and collagen synthesis in VSMC. PMID- 10452124 TI - IL-10 and trichosanthin inhibited surface molecule expression of antigen processing cells and T-cell proliferation. AB - AIM: To study immunoinhibitory effects and preliminary mechanism of IL-10 and trichosanthin. METHODS: Surface molecule expression on antigen processing cells (APC) was stained with fluorescence and analyzed by FACScan. B7-1 mRNA expression was detected with nested RT-PCR. RESULTS: IL-10 2 mg.L-1 and trichosanthin 10 mg.L-1 inhibited B7-1 molecule expression. By contrast, they had not the same effects on ICAM-1. IL-10 and trichosanthin down-regulated LFA-1 expression, but had no regulatory effect on CD40. IL-10 and trichosanthin dramatically inhibited T-cell proliferation and IL-2 production. B7-1 mRNA expression was undetectable in APC treated with IL-10 and trichosanthin. CONCLUSION: IL-10 and trichosanthin inhibit surface molecule expression on APC. They exert multiple immunoinhibitory effects. PMID- 10452125 TI - Demethylation capacity of human fetal adrenal mitochondrial cytochrome P-450 in vitro. AB - AIM: To explore the capacity and characteristics of adrenal mitochondria to metabolize xenobiotics in vitro in human fetus. METHODS: Subcellular fractions of fetal adrenal were prepared by differential centrifugation. Mitochondrial P-450 system was proved by spectral analyses and SDS-PAGE. The formaldehyde formation contents were measured with Nash reagent. RESULTS: The erythromycin N demethylation linearly increased in the protein concentration (1-4 mg)- and incubation time (10-30 min)-dependent manners. A typical concentration-effect relationship appeared with erythromycin 0.067-1 mmol.L-1 and a positive correlation (r = 0.641, P < 0.05) existed between erythromycin N-demethylation and gestation months. The N-demethylation values (nmol.s-1/g protein) of erythromycin (2.7 +/- 0.8), benzfetamine (1.1 +/- 0.5), and aminophenazone (0.9 +/- 0.4) in mitochondria were 89% (P > 0.05), 162% (P < 0.01), and 62% (P < 0.01), respectively, of those in microsomes. There was correlation between mitochondria and microsomes in the N-demethylation of erythromycin (r = 0.708, P < 0.05) and benzfetamine (r = 0.707, P < 0.05). Troleandomycin stimulated erythromycin N-demethylation in adrenal mitochondria as well as in adrenal and liver microsomes in vitro. CONCLUSION: Fetal adrenal mitochondria, with multiple P-450 isoforms and greater capacity of demethylation, play a role in drug metabolism during fetal development. PMID- 10452126 TI - Theoretical elucidation of activity differences of five phenolic antioxidants. AB - AIM: To verify the effectiveness of structure-activity relationship (SAR) and theoretical calculation methods for antioxidants. METHODS: Preliminary elucidation on the differences of activities of 5 antioxidants was performed by SAR. Then semiempirical quantum chemistry method AM1 was employed to calculate the delta HOF value, the difference between the heat of formation of antioxidant and its free radical, which was used as a theoretical parameter to elucidate the differences of activities of the antioxidants thoroughly. RESULTS: delta HOF values of antioxidants were obtained as follows: ferulic acid, 150.58 kJ.mol-1; anion of ferulic acid, 122.64 kJ.mol-1; modified ferulic acid, 137.70 kJ.mol-1; anion of modified ferulic acid, 118.99 kJ.mol-1; salvianic acid, 134.17 kJ.mol-1; rutin, 137.83 kJ.mol-1, L-EGCG, 124.39 kJ.mol-1; paeonol, 176.79 kJ.mol-1. The differences of the antioxidant activities were elucidated, and how to further enhance the antioxidant activity was investigated as well. CONCLUSION: The SAR and calculation methods are rather effective to elucidate the differences of antioxidant activities, and present some new clues for structural modification of antioxidants to increase their activities. PMID- 10452127 TI - Pharmacokinetics of atenolol enantiomers in 12 Chinese healthy men. AB - AIM: To study the pharmacokinetics of atenolol (Ate) stereoisomers in Chinese. METHOD: A single oral dose of 100 mg of racemic Ate tablets were given to 12 healthy volunteers of Han nationality. Plasma and urine concentrations were determined by the reversed phase HPLC method. RESULTS: The disposition of d-Ate and l-Ate was conformed to one-compartment model. Maximal plasma concentration (Cmax): l-Ate (331 +/- 79) micrograms.L-1, d-Ate (342 +/- 78) micrograms.L-1. Area under blood concentration-time curve (AUC): d-Ate (2635 +/- 610) micrograms.h.L-1, l-Ate (2442 +/- 588) micrograms.h.L-1. Renal clearance (Clr): l Ate (6.9 +/- 1.2) L.h-1, d-Ate (6.5 +/- 1.3) L.h-1. CONCLUSION: The disposition of Ate stereoisomers is of stereoselectivity. PMID- 10452128 TI - Tissue distribution of recombinant human tumor necrosis factor alpha derivative in mice. AB - AIM: To study the tissue distribution and its mechanism of a new recombinant tumor necrosis factor alpha derivative (rhTNF alpha Da) in mice. METHODS: 125I rhTNF alpha Da was prepared by Iodogen method. Tissue distribution of 125I-rhTNF alpha Da in mice was studied by determining radioactivity of tetrachloroacetic acid (TCA)- precipitable fraction in tissues. The isolated heart-lung perfusion study using 125I-rhTNF alpha Da perfusate was carried out to study the distribution characteristics of 125I-rhTNF alpha Da in lung. RESULTS: Except for thyroid, AUC of the TCA-precipitable 125I-rhTNF alpha Da in tissues was highest in lung, which was 12.2-fold of that in serum, while concentrations in other tissues were all lower than that in serum. Perfusion study in vitro revealed that the concentration of radio-labeled peptide in lung was higher than that in perfusate. On the contrary, level in heart was much lower than that in perfusate. The overall distribution of 125I-rhTNF alpha Da in lungs showed rapidly equilibratory, dose-dependent, saturable, competitive, and highly affinitive, with Kd 47.6 pmol.L-1 and Bmax 348 fmol.g-1 (lung tissue). CONCLUSION: The specific distribution of rhTNF alpha Da in lungs was its distinctive characteristics. PMID- 10452129 TI - Correlation between inhibitions of morphine withdrawal and nitric-oxide synthase by agmatine. AB - AIM: To study correlation between inhibitions of naloxone-precipitated withdrawal jumps and nitric-oxide synthase (NOS) activity by agmatine. METHODS: NOS activities in mouse brain were measured by determination of concentration of [3H]citrulline, the product of [3H]arginine. RESULTS: Agmatine inhibited NOS activity in naive and morphine-dependent mouse cerebellum, forebrain, and thalamus in substrate-competitive manner in vitro. Naloxone induced withdrawal jumps and an increase in NOS activity in cerebellum, forebrain, and thalamus of abstinent mice. Pretreatment of mice with morphine plus agmatine inhibited the effect of naloxone to precipitate withdrawal jumps and increase in NOS activity. The effect of agmatine was blocked by idazoxan. CONCLUSION: The inhibitory effect of agmatine on naloxone-precipitated withdrawal jumps is related to its inhibition of NOS activity by substrate competitive manner and activation of imidazoline receptors. PMID- 10452130 TI - Dihydroetorphine-induced place preference was mediated by dopamine D1 receptors in rats. AB - AIM: To study the influence of dopamine (DA) receptor antagonists upon the rewarding property of dihydroetorphine (DHE). METHODS: Conditioned place preference (CPP) paradigm was used to characterize the rewarding effect of DHE. DA receptor antagonists were injected administered subcutaneously or peritoneally and microinjected into nucleus accumbens (NAcc). RESULTS: DHE (0.05, 0.5, and 5.0 micrograms.kg-1, s.c.) produced place preference (P < 0.01). Both the DA receptor antagonist haloperidol and the selective D1 receptor antagonist Sch-23390 attenuated the place preference produced by DHE (0.5 microgram.kg-1, s.c.). l Sulpiride and spiperone, selective D2 receptor antagonists, had no such effects. CONCLUSION: The D1 (but not D2) receptors in NAcc are crucial in the mediation of the rewarding effect of DHE. PMID- 10452131 TI - Making the quality connection. PMID- 10452132 TI - Strategic risk management: reducing malpractice claims through more effective patient-doctor communication. AB - Many malpractice suits are brought not because of malpractice nor even because of complaints about the quality of medical care but as an expression of anger about some aspect of patient-doctor relationships and communications. The theory presented is that under the stress of anxiety and physical illness, some patients regress to childhood needs; physicians are not generally trained to fill such needs. Thus, these patients, angry because of this, express their anger in malpractice suits. This theory has been taught to physicians and medical students as part of a physician continuing medical education (CME) seminar on Loss Prevention/Risk Management through demonstration of active-listening techniques to seminar participants. Physicians who understand and can respond appropriately to the emotional needs of their patients are less likely to be sued. This may also translate into a more fulfilled practice of medicine by those physicians who are most aware of the importance of a positive relationship. PMID- 10452133 TI - What is necessary for high-quality discharge summaries? AB - The objective of this study was to determine what physicians perceive to be necessary for high-quality discharge summaries. One-on-one surveys of 100 hospital-based physicians-in-training and community family physicians were conducted. Participants indicated the amount that 56 items contributed to discharge summary quality on a 15-category ordinal scale. Results were transformed to a continuous scale, extending from -6.6 ("item makes summary useless") through 0 ("item has no effect on discharge summary quality") to 10 ("item is so essential that summary is useless without it"). Quality decreased significantly when summary length exceeded 2 pages and when the delay from patient discharge to summary delivery increased. Summary content that increased quality most included admission diagnosis (mean 8.2; 95% confidence interval [7.7, 8.6]), pertinent physical examination findings (6.6 [6.0, 7.2]) and laboratory results (6.8 [6.3, 7.4]), procedures (7.1 [6.7, 7.6]) and complications in hospital (7.1 [6.6, 7.5]), discharge diagnosis (8.8 [8.4, 9.1]), discharge medications (7.9 [7.4, 8.4]), active medical problems at discharge (7.8 [7.4, 8.2]), and follow up (6.6 [6.0, 7.1]). With minor exceptions, hospital and family physicians agreed on contributors to summary quality. For this sample of physicians, summaries were of high quality when they were short, delivered quickly, and contained pertinent data that concentrated upon discharge information. PMID- 10452134 TI - Evaluation of physician preferences for guideline implementation. AB - Although current literature supports the use of evidence-based clinical practice guidelines (CPGs) by physicians, there is limited research concerning operational issues that may be inhibiting effective CPG implementation. The objective of our research was to increase understanding of clinical practice patterns by identifying physician preferences for CPG accessibility, format, content and learning strategies. Semistructured interviews were conducted with resident and faculty physicians in an academic medical center after they were presented with a CPG during treatment of a patient with acute pancreatitis. The results of our study revealed that physicians prefer CPGs in the form of evidence-based algorithms with treatment-specific information that is placed on the front of the patient chart during treatment. In addition, they felt that discussion of the guideline with colleagues, reminder notes/stickers on front of the patient chart, and verbal reminders from a nurse were the most effective means of encouraging utilization. PMID- 10452135 TI - Incorporating results of a provider attitudes survey in development of an outcomes assessment program. AB - The objective of this study was to obtain information from providers of a behavioral health service and from decision makers for organizations interacting with that service (external contacts) on their attitudes regarding outcomes assessment in their clinical practice. The goal of obtaining the information was to use it in development of a formal Outcomes Assessment Program for the service. The design was a semi-structured interview format, with entry into a computer database and qualitative analysis of responses obtained. Participants included all providers (n = 26) and a purposive sample of external contacts (n = 10) of an academic Department of Psychiatry. Results indicated differences among categories of external contacts regarding priorities of types of outcomes (general health, general mental health, disorder specific, or patient satisfaction) to be shared and absence of concordance within the service about these priorities. No guidelines were available about preferred instruments, though the Global Assessment of Functioning, the Beck Depression Inventory, and the Abnormal Involuntary Movements Scale emerged as instruments to be prioritized in the service's program. Physicians and nonphysicians differed in their perceptions as to important barriers and advantages of a clinical outcomes assessment program. In conclusion, the survey raised providers' awareness of outcomes assessment and provided information that was used in developing the service's Outcomes Assessment Program. Components of the Program that were influenced by survey input were priorities of outcomes instruments to be included and their potential audiences, time sequence of Program development, time to be allotted to outcomes assessment in clinical encounters, and content of educational experiences for providers. PMID- 10452136 TI - Platelet availability at North Iowa Mercy Health Center: a CQI positive result. AB - Physicians complained because outpatients had to wait for platelet transfusions because of product unavailability. Platelet use had increased 30%, and additional platelet shipments from our supplier were often required. A Continuous Quality Improvement (CQI) committee identified 2 positive impact factors: adjust inventory stocked and recommend a 4-unit dose for cardiac surgery, on the basis of supplier data validating that platelets in 4 concentrates exceeded the minimum apheresis standard. Subsequent to these changes, all orders have been filled immediately, and the average platelet use in cardiac surgery has decreased from 7 to 4 units. There have been no instances of inability to fill platelet orders. Currently, 96% of cardiac surgery platelet orders are for 4 units. Prior to the adjustment to inventory, 24 requests in 6 months were placed with our supplier for additional platelet deliveries. Requests have decreased to 3 in 6 months. PMID- 10452137 TI - [Should we always treat partial benign epilepsy in childhood?]. PMID- 10452138 TI - [Repercussions in children of drug addicted parents]. PMID- 10452140 TI - [Spirometric values in healthy children of the urban area of the Basque Autonomic Community]. AB - OBJECTIVE: Our objective was to obtain spirometric reference values for children. PATIENTS AND METHODS: We performed a transversal study in schools in Bilbao of children of both sexes that were between 7 and 14 years of age, non-smokers and healthy (according to a questionnaire and physical examination). A sample of 657 subjects was calculated based on the standard deviations of height for each sex and age group. The sample units were the schools and they were selected randomly. The following data were collected: age, sex, weight, height, FVC, FEV1, FEV1/FEC, PEF, FEF25, FEF50, FEF75 and FEF25-75. Forced spirometry was carried out according to the ATS guidelines by using a MasterScreen model 4.10.b (Jaeger, Germany). A descriptive statistical study was performed, as well as a normality test and linear regression. RESULTS: We studied 765 subjects (415 boys and 350 girls). The natural logarithm of the spirometric variables was taken to obtain a Gaussian distribution. These variables were related to age, weight and height. The relationship improved when associated with sex, with the best association being between height and sex. The equations obtained were only valid for FVC and FEV1, and were as follows: in boys FVC(L) = -1.968 + 0.020 height and FEV1(L) = 1.831 + 0.018 height and for girls FVC(L) = -1.879 + 0.019 height and FEV1(L) = 1.809 + 0.018 height [R2 = 0.893, 0.891, 0.868 and 0.871, respectively]. CONCLUSIONS: The pediatric reference values for forced spirometry were established for our population, complying with the current applicable guidelines. PMID- 10452139 TI - [Nutritional anthropometry and physical performance in urban adolescents of Madrid]. AB - OBJECTIVE: This work has been carried out as a cooperation project between University and High School Departments. Its objective was to evaluate the nutritional and physical status and eating habits in an urban adolescent population of Madrid. The study presents data about anthropometrical analyses and physical performance. SAMPLE AND METHODS: The sample analyzed was composed of 183 males and 222 females between 14 and 18 years of age. The nutritional status was analyzed by means of stature, weight, skinfold thickness and arm circumference, body mass index (BMI), and lean, fat and total upper arm areas. For physical fitness analysis, the EUROFIT battery was used (plate tapping, trunk and arm flexion, 10 x 5 run, horizontal jump, abdominals in 30 seconds and manual dynamometry). RESULTS AND CONCLUSIONS: With regard to the anthropometric characteristics and physical fitness, the analyzed population is slightly above the reference standard, mainly in the youngest age studied. Percentage of obese subjects was 14.7% in males and 5.8% in females. In regard to physical fitness, the males surpassed the 50th percentile of the EUROFIT battery in almost all of the trials. The average in the female subjects was slightly inferior to the reference standards. PMID- 10452141 TI - [Liver complications associated with short-term parenteral nutrition in children]. AB - OBJECTIVE: Liver dysfunction (LD) with abnormalities in biochemical liver function tests is the most common metabolic complication of parenteral nutrition (PN). The aim of this study was to estimate the prevalence of LD in children receiving short-term PN and to identify risk factors. PATIENTS AND METHODS: Data were gathered retrospectively during a 2-year period. Ninety-four children older than 28 days received PN (mean age 5.4 +/- 5.1 years). PN related LD was defined as when serum levels of one or more of the following liver function tests were increased: ALT > or = 80 IU/L, GGT > or = 120 IU/L and total bilirubin > or = 1.8 mumol/L. Children with previous liver disease were excluded (n = 17), as well as those with incomplete data (n = 16). RESULTS: LD was present in 33 children (54%). The incidence rate was 5.8 cases/100 patient days of PN. It started 9.8 +/ 6.9 days after beginning PN. The nadir appeared during the second week of PN. The following variables did not appear to significantly influence the presence of PN: age, gender, nutritional status, PN caloric load or composition and underlying disease. Length of PN (9.6 +/- 4.4 vs 19.5 +/- 10.5 days; p < 0.001) and presence of sepsis (21% vs 55%, p = 0.014) were the only variables associated with LD. It was not necessary to discontinue PN because of LD in any case. CONCLUSIONS: Early LD is present in more than 50% of our children on PN. In preventing LD we should try to avoid infection and to reduce the time on PN. PMID- 10452142 TI - [Inadequate secretion of atrial natriuretic peptide in children with acute brain injury]. AB - OBJECTIVE: Fluid and electrolyte disturbances are frequent after acute brain damage. Atrial natriuretic peptide (ANP), which can also be found in brain tissue, could be a hormone implicated in such disorders (cerebral salt wasting syndrome). PATIENTS AND METHODS: Plasma ANP levels were analyzed in 50 children with acute neurological deterioration (secondary to traumatic, infectious, convulsive tumor or vascular disorders) evaluated according to the modified Glasgow Coma Scale (GCS). Hemodynamic stability was determined by standard monitoring. ANP, renin and aldosterone levels were determined by RIA and the results compared with a control group of healthy children. RESULTS: There was an increase in ANP levels in children with brain injury in comparison to the control group (p < 0.001), but not in children under one year of age. Renin and aldosterone were also significantly increased in this group of patients, with no difference in ANP, renin or aldosterone level found in function of their GCS score. All patients were hemodynamically stable and no correlation between hemodynamic and hormone variables was seen. Mechanical ventilation did not influence the hormone levels. CONCLUSIONS: There is an important increase in ANP levels in patients with acute neurological pathologies, but it was not related to the hemodynamic condition and its importance has yet to be established. One of its possible consequences is a secondary hypotonic/hypovolemic condition, a potentially dangerous event for the patient with intracranial hypertension that needs immediate treatment. Differentiation of this syndrome from inappropriate vasopressin secretion could be very important in children with acute brain injury. PMID- 10452143 TI - [Gastric emptying in children with gastroesophageal reflux. Evaluation by real time ultrasonography of the pyloric antrum]. AB - OBJECTIVE: We have studied gastric emptying using real time ultrasound in 25 healthy children and 25 children with RGE. Our aim was to establish the time and mode of gastric emptying and the volume and the degree of antral dilation in both groups at baseline and after a test meal. We also evaluated the same parameters in the RGE cases after supplying a prokinetic. PATIENTS AND METHODS: The Bolondi technique was used to measure the basal antrum volume and emptying ratio, as well as the mode and final gastric emptying time at different time-points after a solid-liquid test meal. RESULTS: Patients with RGE have a clearly longer gastric emptying time with respect to the healthy subjects in both children less than one year of age (238 +/- 48.4 vs 176.3 +/- 36.7 minute; p = 0.03) and those 1-14 years old (206.3 +/- 48.2 vs 163.3 +/- 48.1 minutes, p = 0.03). An anomalous gastric emptying, similar to a broken line, can be observed in children suffering from RGE. In contrast, a progressive reduction of antrum dilation ration was seen in healthy children and not in patients with RGE. After cisapride, gastric volume goes down at all test times in RGE children. Therefore, a decrease in final gastric emptying time cannot be observed. CONCLUSIONS: Children suffering from RGE show an alteration in gastric emptying with a clearly greater time and an anomalous mode of emptying. The antrum remained dilated for longer periods after ingestion of test food. PMID- 10452144 TI - [Why is it preferable to use air in the radiological reduction of intestinal intussusception?]. AB - OBJECTIVE: We present our experience describing the radiological findings in the treatment of intestinal intussusception. PATIENTS AND METHODS: A retrospective study of the clinical and ultrasound findings and their treatment in 131 intussusceptions diagnosed in 126 patients was performed. RESULTS: In 102 cases, the pneumatic intussusception reduction under radiological control was possible. Twenty-nine patients underwent surgery and amongst these the pathological changes in the intestinal wall indicated resection in 9. CONCLUSIONS: The pneumatic intussusception reduction under fluoroscopic monitoring is now more accepted than hydrostatic reduction with barium. With the proper equipment and experienced staff, this is the most promising method in the management of intestinal intussusception during infancy. PMID- 10452145 TI - [Lipoprotein (a) and family history in 6-year-old children from Rivas Vaciamadrid]. AB - OBJECTIVE: The purpose of this study was to evaluate levels of lipoprotein(a) as a marker of family history of cardiovascular risk. PATIENTS AND METHODS: We have studied 673 six-year old children. Lipoprotein(a) was determined and previous family history of cardiovascular risk (hypercholesterolemia and cardiovascular disease under 55 years) was collected. RESULTS: Of the children studied, 7.42% had positive antecedents of cardiovascular risk in the parents, 51.8% in grandparents and 9.21% in parents and grandparents. The lipoprotein(a) levels were higher in these groups than in those without any family history of cardiovascular risk. Likewise, the percentage of children with lipoprotein(a) levels higher than 30 mg/dl was 15.69% in the group without a family history of problems and 25.8% and 25.69%, in the group with hypercholesterolemia antecedents and cardiovascular disease, respectively. The sensitivity, specificity and positive predictive value test did not prove that a positive family history could be used to detect the majority of children with lipoprotein higher than 30 mg/dl. CONCLUSIONS: Lipoprotein(a) levels in children are higher in those with a family history of cardiovascular disease or hypercholesterolemia, but these antecedents cannot be used as a determinant factor to detect the majority of children with lipoprotein(a) higher than 30 mg/dl. PMID- 10452146 TI - [The delay of the diagnosis of deafness in children. Justification for the establishment of screening models]. AB - OBJECTIVE: Our objective was to assess the capacity of our system of primary care to diagnose hearing loss in children in order to prevent isolation behaviors and obtain their acceptable insertion into our society. PATIENTS AND METHODS: A review of 78 cases of children, between 0 and 14 years of age, with hearing loss and followed in our Audiology Department was made. The age in which the diagnosis was made, presence of risk markers for deafness, records and motive of consultation were analyzed. RESULTS: The mean age at diagnosis was 21 months. Although there were risk markers in 45 children (57%), screening tests, mainly brainstem auditory evoked potentials, were performed in only 7. Moreover, the main motive for consultation was the lack of response to sound stimuli as observed by the parents, and not the presence of any risk marker. CONCLUSIONS: Better information for parents and tutors and a greater conscientiousness of clinicians towards the possibility of deafness in children, especially in risk populations, would reduce the age at which the deficit is detected. The availability of brainstem auditory evoked potential tests in primary care centers will allow the diagnosis to be made in most children before 12 months of age. PMID- 10452147 TI - [Etiology of central diabetes insipidus in children]. AB - OBJECTIVE: The aim of this report were to document the etiology of central diabetes insipidus (CDI) in children under fourteen years of age and to describe the time of onset of CDI after its cause. PATIENTS AND METHODS: A descriptive retrospective study using the records of 33 children diagnosed of CDI between January 1988 and December 1997 was performed. RESULTS: Brain death produced CDI in seventeen patients, intracranial tumors in five (after excision of the tumor) and seven had other causes. No etiology was detected in four. Time interval from the cause, when known, to the onset of CDI was always less than seven days. CONCLUSIONS: The most frequent etiology of CDI in children was brain death (51%). In all of our patients, the time of onset of CDI was less than one week. PMID- 10452148 TI - [Bone marrow transplantation in utero]. PMID- 10452149 TI - [Neurological and growth follow-up of cardiac patients operated on before 45 days of life]. AB - OBJECTIVE: Our aim was to study the neurological and growth disorders in patients undergoing cardiac surgery during the neonatal period. PATIENTS AND METHODS: Ninety clinical records of newborns with congenital heart disease that were operated between 1985 and 1996 and were under the age of 45 days with a minimum follow-up period of 12 months were reviewed. RESULTS: Of the 37 patient records considered, 33 (89.1%) were neurosensorially normal, 2 had minor handicaps and 2 moderate handicaps. There were 15 (40.5%) with the weight and 9 (24.3%) with the length below the 10th percentile. Although exitus, handicaps and growth disorders were more frequent in those patients operated in the extracorporal circulation, there were no significant differences. The growth disorders were greater in complex heart diseases and in pulmonary atresia, but significant differences were only related to the weight. CONCLUSIONS: Because at least 11% of these patients showed neurological disorders, a neurological follow-up is necessary. The number of post-operated patients with growth disorders was high, but catch-up growth resulted in recovery. A greater number of neurological and growth disorder has been found in complex heart disease, where only a palliative treatment could be employed. Although the complete correction of the cardiopathy may imply an early mortality, long time prognosis for survivors is much better. PMID- 10452150 TI - [Hepatic fascioliasis. A new case during childhood]. PMID- 10452151 TI - [Radial club-hand and contralateral pre-axial polydactyly and ulnar club-hand and contralateral post-axial polydactyly. Report of 2 unusual cases]. PMID- 10452152 TI - [Cerebral abscess due to Streptococcus intermedius resistant to metronidazole: a report of 3 cases]. PMID- 10452153 TI - [Microcytic anemia resistant to the oral administration of iron as a form of presentation of the Castleman disease]. PMID- 10452154 TI - [Recurrent post-prandial hypoglycemia: reactive hypoglycemia during childhood? The use of acarbose]. PMID- 10452155 TI - [Membranous atresia of the colon]. PMID- 10452156 TI - [Vocal cord dysfunction in an asthmatic child]. PMID- 10452157 TI - [Transient neonatal diabetes mellitus: a report of two cases]. PMID- 10452158 TI - [What is your diagnosis? A 2-month-old infant with respiratory distress]. PMID- 10452159 TI - [Epidemiology and scientific methodology applied to pediatrics (VI): confusion and interaction]. PMID- 10452160 TI - [Advances in neonatology]. PMID- 10452161 TI - Diagnosis and management of teeth with vertical root fractures. AB - Vertical fractures in teeth can present difficulties in diagnosis. There are, however, many specific clinical and radiographical signs which, when present, can alert clinicians to the existence of a fracture. In this review, the diagnosis of vertical root fractures is discussed in detail, and examples are presented of clinical and radiographic signs associated with these fractured teeth. Treatment alternatives are discussed for both posterior and anterior teeth. PMID- 10452162 TI - A brief review of indigenous Australian health as it impacts on oral health. AB - The indigenous population of Australia constitutes approximately 2 per cent of the total population. This group has faced significant cultural, economic and health changes since European settlement some 200 years ago. In this brief review some of the health changes that have influenced the oral health status of this community have been examined. Of major importance is the dietary change that the once nomadic indigenous community has undergone. Today's Western diet, high in sugar, low in proteins and vitamins, has resulted in a significant increase in the risk (and prevalence) of caries and periodontal disease. In addition, the high prevalence of diabetes also exacerbates the periodontal problem. The remoteness of a significant proportion of Australian indigenous communities from modern health care services and limited access to fluoridation increases the incidence of oral disease. It is also noted that the incidence of rheumatic heart disease is one of the highest in the world, thereby increasing the risk of bacterial endocarditis. It is clear that indigenous communities have unique oral health needs but the extent of these needs is not well documented. It is important that more research be undertaken to assess these needs so that appropriate oral health programmes can be developed. PMID- 10452163 TI - Oral cancer: role of the basement membrane in invasion. AB - Invasive growth of cancer cells is a complex process involving specific interactions between tumour cells and the orderly, integrated complexes of the extracellular matrix. Basement membranes have been proposed as one constituent of extracellular matrix which carries responsibility for regulating invasion and metastasis. Using a chemically induced rat tongue carcinoma model, it has been shown that components of the basement membrane and its overall structure are altered during tumour invasion, and methods have been developed to quantitate some of these differences. Since the basement membrane can be specifically characterized by its fibrous protein network of Type IV collagen and laminin, which is embedded in a heparan sulphate-rich proteoglycan matrix, these components have been targeted. In particular, the current paper presents results in the context of current concepts of early changes in neoplastic invasion of underlying connective tissues. In consequence, further elaboration of the underlying mechanisms of epithelial migration in oral cancer may allow an exploration of the use of alterations in expression of basement membrane components as prognostic indicators. PMID- 10452164 TI - The effects of the Nd:YAG laser on amalgam dental restorative material. AB - The Nd:YAG laser has been marketed as an instrument for use on both hard and soft dental tissues. Its potential for use on hard tissues is limited but it may be the instrument of choice for use in certain soft tissue procedures. The aim of this study was to examine the effects of the Nd:YAG laser on amalgam restorations which are frequently placed on tooth surfaces adjacent to areas of soft tissue which may be subjected to the laser. The amalgam used was Tytin. The laser firing was controlled by a computer and a constant repetition rate of 40 Hz was used. Energy per pulse was altered as follows: 30 mJ, 40 mJ, 60 mJ, 80 mJ, 120 mJ and 140 mJ. Exposure times of 0.05 s (2 pulses), 0.125 s (5 pulses), 0.25 s (10 pulses), 0.5 s (20 pulses), 1 s (40 pulses), 2 s (80 pulses), 3 s (120 pulses), 4 s (160 pulses), and 5 s (200 pulses) were used. The width of defect was measured using a measuring microscope with 10x magnification and it was established that the damage threshold lies between 0.125 s (5 pulses) and 0.25 s (10 pulses) for 30 mJ per pulse. Scanning electron microscope observations revealed that the melting of amalgam at exposure times of 5 s actually decreased the size of the observed defect. The data were analysed using a two-way ANOVA statistical test. There was a significant (p < 0.001) correlation between the width of the defect and exposure time up to an exposure time of 4 s and the width of defect and the energy per pulse setting. The findings indicate that amalgam restorations are damaged by inadvertent laser exposure and clinicians must take measures to protect such restorations during lasing of soft tissues. PMID- 10452165 TI - p53 expression in oral precancer and cancer. AB - Expression of the p53 tumour suppressor gene is a frequent finding in human malignancies, including oral cancer, and it has been detected in some potentially malignant lesions. The results of the present project showed that 35 of the 41 (85 per cent) oral mucosal lesions with histological evidence of epithelial dysplasia expressed p53, but the presence or absence of p53 staining could not be used to predict the outcome of potentially malignant oral mucosal lesions. PMID- 10452166 TI - The outcome of arthroscopic treatment of temporomandibular joint arthropathy. AB - Ninety patients underwent arthroscopic temporomandibular joint surgery to 124 joints for arthropathy which had failed to respond to at least six months of non surgical treatment. They were surveyed at between 6 months and 5 years (mean 2.5 years) after surgery and 63 per cent responded to the survey. They reported an 82 per cent improvement for pain (50 to 100 per cent better), 80 per cent for clicking and 82 per cent for locking. There was no morbidity following the treatment. Arthroscopic surgery should be considered for advanced temporomandibular joint arthropathy which is refractory to non-surgical treatment. PMID- 10452167 TI - Root resorption of mandibular second molar teeth associated with the presence of the third molars. AB - Although pressure resorption of second molars is associated with impacted and/or erupting third molars, the relationship between pressure resorption and age is unclear. In order to investigate this relationship, 3883 dental radiographs were studied in 3174 individuals of various ages. The incidence of superimposition of the second molar on the third molar was not elevated in any subgroup except that of women of 16-40 years of age. There were no age or sex differences for the incidence of second molar root resorption. In older individuals, root resorption associated with a completely impacted third molar was more frequent than with a partially impacted third molar, and root resorption at the apex was mainly seen in individuals over 50 years of age. There was a higher incidence of superimposition and root resorption in men than women. Apical root resorption may be seen long after the formation of completely impacted third molars in both genders. PMID- 10452168 TI - Radiographic appearance of alveolar osseous defects in relation to their anatomic location. AB - Radiographs are of limited value in the diagnosis of osseous defects. Anatomic and technical factors affect the radiographic appearance of bone lesions. This study was undertaken in order to determine whether the radiographic appearance of alveolar osseous defects depends on their location, and particularly whether there are any differences in radiographic detection between bone defects located in the maxilla or the mandible, and on the buccal or lingual aspects of the alveolar crest. Experimental bone lesions were created in the alveolar crest of a skull. Standardized periapical radiographs were obtained before and after the defects were made. After processing, pairs of radiographs were randomly mounted. Five dentists acted as observers in order to determine whether or not a change in alveolar bone was detectable at each of the five possible locations. The unpaired t test was applied for the assessment of statistically significant differences with respect to sensitivity, specificity, and positive diagnostic value in the diagnosis of bone lesions depending on their location. The results showed that the anatomic location of a lesion in the alveolar bone affected its radiographic appearance. Moreover, experimental defects were detected more often in the mandible and on the lingual surfaces of the alveolar crest. PMID- 10452170 TI - Central fibroma in the ascending ramus of the mandible. Case report. AB - A case of central fibroma involving the mandible in a 58 year old woman is described. There was slight swelling of the left cheek and bone-hard bulging was detected on palpation but the patient had not complained of the swelling. The lesion was removed under general anaesthesia and then examined histopathologically. There was no sign of recurrence eleven months after the operation. PMID- 10452169 TI - The effects of extracellular citric acid acidosis on the viability, cellular adhesion capacity and protein synthesis of cultured human gingival fibroblasts. AB - Root surface demineralization is widely used as an adjunct to periodontal treatment. To clarify the influence of citric acid root conditioning on periodontal wound healing, the effects of citric acid and associated extracellular acidosis on the viability (MTT assay), attachment and protein synthesis ([3H]-proline incorporation into trichloroacetic acid-precipitated proteins) of human gingival fibroblasts (GF) were investigated. A concentration of 47.6 mmol/L of citric acid (pH 2.3) in water led to total cell death within three minutes of incubation. Media containing 23.8 mmol/L and 47.6 mmol/L of citric acid exerted strong cytotoxicity (47 to 90 per cent of cell death) and inhibited protein synthesis (IC50 = 0.28 per cent) of GF within three hours of incubation. Incubation of cells in a medium containing 11.9 mmol/L of citric acid also suppressed the attachment and spreading of fibroblasts on culture plates and Type I collagen, with 58 per cent and 22 per cent of inhibition, respectively. Culture medium supplemented with 11.9, 23.8 and 47.6 mmol/L of citric acid also led to extracellular acidosis by decreasing the pH value from 7.5 to 6.3, 5.2 and 3.8, respectively. In addition, it was confirmed that the toxic effect of media containing citric acid was due to their acidity rather than the citrate content. Most of the citric acid-induced cell death could be prevented by adjusting the pH value of the culture medium to pH 7.5. Sodium citrate, at a concentration of 47.6 mmol/L, also exerted little cytotoxicity. The results suggested that toxicity of citric acid in specific stages of the healing process must be considered prior to its clinical application. Careful management of citric acid in order to avoid contact with tissue or the development of other demineralizing agents is important in enhancing periodontal wound healing. PMID- 10452171 TI - Adenomatoid hyperplasia in the mandibular retromolar area. Case report. AB - This paper reports a case of adenomatoid hyperplasia of minor salivary glands in the retromolar area, with a unique symptom of painful swelling. This appears to be the first reported case in the Chinese population. It would be useful for clinical dentists to be aware of this tumour. PMID- 10452172 TI - Restorative materials. PMID- 10452173 TI - Anxiety sensitivity and fear of pain in patients with recurring headaches. AB - Anxiety sensitivity (AS) plays an important role in the cognitive, affective and behavioral profiles of patients with chronic pain related to musculoskeletal injury. However, investigators have not considered whether these findings extend to patients with other classes of chronic pain. The primary purpose of this investigation was to address this issue in 72 patients with recurring headaches who completed a self-report questionnaire inventory during a treatment visit to an outpatient neurology clinic. The mean ASI score for the group (mean = 24; SD = 11) was relatively high. When patients were classified on the basis of ASI scores, 20 (28%) met criteria for high, 41 (57%) for medium and 11 (15%) for low AS. Multivariate analysis of variance confirmed that these groups differed on specific aspects of their cognitive, affective, and behavioral profiles. High AS patients reported greater depression, trait anxiety, pain-related escape/avoidance behavior and fearful appraisals of pain than did patients with medium or low AS. High AS patients also indicated greater cognitive disruption in response to pain than did patients with low AS. Groups did not differ in headache severity, physiological reactivity, change in lifestyle, anger, nor did they differ in use of over-the-counter or prescribed analgesics. Multiple regression analysis identified AS, pain-related cognitive disruption, and sensory pain experience as significant predictors of fear of pain. Lifestyle changes attributed to headache were, on the other hand, predicted by headache severity, physiological and cognitive anxiety and escape/avoidance behavior. These results provide further evidence of the important association between AS and fear responses of patients with chronic pain syndromes. Implications and future directions are discussed. PMID- 10452174 TI - Treatment of childhood memories: theory and practice. AB - With the growing interest of cognitive behaviour therapy in early developed psychopathology like personality disorders there is an increased need for therapeutic methods for more directly treating pathogenic schemas. Exploring and reinterpreting memories of early childhood experiences that are assumed to have contributed to the pathogenesis are more and more viewed as a promising way to modify core schemas. Experiential methods seem to be the most effective. This article discusses two main forms of these methods: (i) imagery with rescripting and (ii) role play, both of childhood interactions with key figures. For both, protocols are provided as guidelines for clinicians and to stimulate standardization so that this new field can be opened for experimental research. Theoretical views are discussed as to why these methods might be so effective in treating chronic problems that originated in childhood. PMID- 10452175 TI - Psychometric appraisal of the Scale for Interpersonal Behavior (SIB) in France. AB - The present study was carried out in France to evaluate the reliability and validity of the Scale for Interpersonal Behavior (SIB), a multidimensional measure of difficulty and distress in assertiveness that was originally developed in The Netherlands. This appraisal was conducted with a clinical sample (N = 166) and a general population sample (N = 150). The clinical series comprised 115 patients with social phobia and 51 patients with personality disorder, 28 of whom were of the avoidant type. Support was found for internal consistency and test retest reliability of the French SIB. Compared to controls, both social phobics and patients with an avoidant personality disorder had significantly lower mean scores on all performance scales and significantly higher ones on all distress scales, with the social phobics occupying a position in between. Findings in relation to convergent and divergent validity were quite satisfactory. Sensitivity of the French SIB for detecting change was demonstrated in a subgroup of the clinical Ss who had undergone 15 sessions of cognitive-behavioral group therapy for underassertiveness. PMID- 10452176 TI - Group cognitive-behavior therapy for patients failing to respond to pharmacotherapy for panic disorder: a clinical case series. AB - The last decade has brought exciting advances to the treatment of panic disorder and agoraphobia, and a variety of cognitive-behavioral and pharmacologic treatment strategies offer clear benefit to patients. Nonetheless, treatment nonresponse continues to be a chronic problem, and additional strategies are needed to aid patients who do not respond fully to initial interventions. In the present study, we use 'services' research to document the clinical response of pharmacotherapy nonresponders to a standard program of brief, group cognitive behavioral therapy. Patients responded well, regardless of whether they had received a full, adequate trial of pharmacotherapy. In addition to its application as an initial treatment for panic disorder, routine application of cognitive-behavioral therapy in pharmacologic treatment algorithms is encouraged, with attention to referral of pharmacotherapy nonresponders to cognitive behavioral therapy. PMID- 10452177 TI - Obsessive-compulsive disorder, anxiety disorders, and self-esteem: an exploratory study. AB - In a preliminary investigation of the link between self-esteem and obsessional problems, patients with OCD were compared with people suffering from other anxiety disorders and non-anxious controls. A questionnaire was devised which allowed the reliable coding of open ended responses focussed on issues surrounding self-worth; standardized measures of self-esteem and clinical symptomatology were also administered. Results indicated that both clinical groups differed significantly from non-clinical controls on generalized self esteem assessments. There was some evidence of OCD specific effects; obsessionals were more likely than anxious controls to link their self-worth to other people and their relationships. They also regarded the possibility of causing harm as likely to result in other people making extreme negative and critical judgements of them; the other groups expected the responses of others towards them to be more lenient. The implications for future research and for treatment of OCD are discussed. PMID- 10452178 TI - Perfectionism and depression symptom severity in major depressive disorder. AB - In recent years it has been recognized that perfectionism is a multidimensional construct and two Multidimensional Perfectionism Scales have been developed and investigated in relative isolation [Frost, R.O., Marten, P., Lahart, C., & Rosenblate, R. (1990). The dimensions of perfectionism. Cognitive Therapy and Research, 14, 449-468; Hewitt, P.L., & Flett, G.L. (1991). Perfectionism in the self and social contexts: Conceptualization, assessment and association with psychopathology. Journal of Personality and Social Psychology, 60, 456-470]. The present study sought to evaluate the association between various dimensions of perfectionism, higher-order personality dimensions, and self and observer rated depressive symptoms in a group of 145 patients with major depressive disorder. Only three of ten perfectionism dimensions (socially prescribed perfectionism, concern over mistakes and self-criticism) displayed medium to large correlations with depressive symptoms, especially self-report symptoms reflecting depressive cognitive distortions. The results are discussed in relation to the specificity of perfectionism dimensions to depression, adaptive versus maladaptive aspects of perfectionism, and in the context of previous research, much of which has relied on college student samples. PMID- 10452179 TI - Single/multi drug use. PMID- 10452180 TI - Simple conscious sedation. PMID- 10452181 TI - Electronic dental discussion lists. PMID- 10452182 TI - Graduate training expectations. PMID- 10452183 TI - Post-operative statistics. PMID- 10452184 TI - Cast appeal. PMID- 10452185 TI - Inequalities in oral health: a review of the evidence and recommendations for action. AB - Reducing inequalities in health has become one of the main health policy issues in the late 1990s. The Labour Government set up an independent inquiry into inequalities in health under Sir Donald Acheson to make recommendations on approaches to reducing health inequalities. This paper reviews the evidence on inequalities in oral health in Britain. Dramatic improvements in dental health in children and young adults have taken place in the past 30 years. The levels of caries in permanent teeth of children is low. Widening inequalities in oral health however exist between social classes, regions of England, and among certain minority ethnic groups in pre-school children. The main social class and minority ethnic differences in dental caries is in pre-school children. Wide district and regional differences also exist in prevalence of caries in young children. The area differences relate very strongly to deprivation. In adults the differences in decay experience is less unequal than in children but there are marked social class inequalities in edentulousness. Dental caries decreased in all social classes in the United Kingdom. The main causes of the inequalities are differences in patterns of consumption of non milk extrinsic sugars and fluoridated toothpaste. Improvements in oral health that have occurred over the last 30 years have been largely a result of fluoride toothpaste and social, economic and environmental factors. Oral health inequalities will only be reduced through the implementation of effective and appropriate oral health promotion policy. Treatment services will never successfully tackle the underlying cause of oral diseases. PMID- 10452186 TI - The psychology of dental patient care. 5. The determinants of dental health attitudes and behaviours. AB - This paper examines the role of psychosocial factors as determinants of health behaviour. It examines the development of dental health attitudes from the patient, professional and societal perspectives. PMID- 10452187 TI - Tooth surface loss. 11. Surgical crown lengthening. AB - Restoration of worn teeth can be made easier by surgical crown lengthening. It improves appearance and facilitates tooth preparation. Anatomical features can limit the height that can be gained. PMID- 10452188 TI - Additional studies on the distribution of stresses during vertical compaction of gutta-percha in the root canal. AB - OBJECTIVE: This study was designed to investigate the effect of certain pathological alterations of the dental structures (diminishing bone support, internal resorption, root perforation, periapical lesion) on stress distribution during root canal filling procedures by the warm vertical compaction technique. DESIGN: The computer stress analyses were done for a maxillary canine tooth model which was based on dimensions recovered from a human cadaveric maxilla scanned by CT. METHODS: The finite element method was used to calculate the stresses generated during root canal filling procedures by warm vertical compaction technique. Patterns of stress distribution associated with various alterations in dental structures were investigated. For this purpose 60 cases were simulated. The hypothetical force of 10 N is taken as a unit representation. For other magnitudes of applied force, the corresponding stresses would be scaled directly because the calculations were made for linear materials. RESULTS AND CONCLUSION: It is found that, when diminishing bone support and internal resorption are concurrently simulated, a marked increase in stress magnitudes occur (maximum von Mises stress 5.37 N/mm2). However, these values still remain much below the most frequently reported tensile strength of dentine (50-100 N/mm2). If dentist's handwork is transformed into equivalent edge tractions on gutta-percha, then stresses in dentine, even when they are corrected for 3-kg applied force, appear to remain below fracture strengths of this material. This result leads us to conclude that when warm vertical compaction technique is skillfully performed and inadvertent undue force is not applied, a premature root fracture in a large rooted maxillary anterior tooth with straight root canal anatomy is not likely to occur, even for the unfavourable conditions simulated in our model. This result, like all results derived from modelling applications, is of course contingent upon agreement between the way in which the clinical operations are performed and the way in which they are mirrored for computer representation. We believe that the approach described here avoids the spurious stresses that have been reported in similar investigations. PMID- 10452189 TI - The prevalence and impact of dental pain in 8-year-old school children in Harrow, England. AB - OBJECTIVE: To assess the prevalence, severity and impact of dental pain. DESIGN: Cross-sectional survey. SETTING: Schools in the London Borough of Harrow, England, in the summer term of 1995. SUBJECTS AND METHODS: The base population were all 2,300 8-year-old school children in Harrow. A cluster sampling of schools based on quotas from all postcode areas in Harrow was used. Data were collected through interviews with the children. MAIN OUTCOME MEASURES: Prevalence of previous toothache; prevalence of toothache in the previous 4 weeks; prevalence of toothache in the previous 4 weeks which resulted in a visit to the dentist, in stopping playing, eating, sleeping, going to school and taking painkillers. RESULTS: Of the 664 sample of children in the participating schools, 589 were interviewed (88.7%). The frequency of previous toothache was 47.5% (95% CI, 44-52) and dental pain caused crying in 17.7% (95% CI, 15-21) of children. 7.6% (95% CI, 5-11) of children had pain in the previous 4 weeks (45 children). Among these 45 children, this recent pain resulted in a visit to the dentist in 41.9% (19 children, i.e. 3.2% of all children), in stopping playing in 26.7% (12 children, i.e. 2.0% of all children), eating in 73.3% (33 children, i.e. 5.6% of all children), sleeping in 31.1% (14 children, i.e. 2.4% of all children) and in going to school in 11.1% (5 children, i.e. 0.8% of all children). CONCLUSION: Toothache in children is a sizeable problem in Harrow and had substantial consequences for children and their guardians. Freedom from disabling dental pain/discomfort is an outcome indicator of oral health and could be used as an explicit goal by dental systems. It is important to note however, that the present study did not assess the extent to which the dental pain was associated with avoidable dental problems as opposed to normal physiological processes. It is important that future work try and separate the prevalence of dental pain caused by physiological from avoidable pathological factors. In addition, future work is needed to assess how effectively and efficiently dental services are responding to people suffering with dental pain. PMID- 10452190 TI - Appraisal of a standardised periodontal referral proforma. AB - AIM: The primary objective was to assess the standard of new-patient referral communications to the Periodontology Department of a teaching hospital and evaluate any differences in referral quality between referrals that used a standardised proforma and conventional letters. Secondary objectives were to evaluate the potential influence on referral performance of age, postgraduate qualifications and country of qualification of the referring practitioners. DESIGN: A retrospective analysis of a randomised sample of all referral letters and proformae received during a nine-month period. SETTING: Letters and referral proformae received by the Periodontal Department of the teaching hospital between 1 January and 30 September 1995 were surveyed. METHODS: An objective Categorisation System for Periodontal Referral Quality (CSPRQ) was devised and tested for inter- and intra-examiner reproducibility. The referral communications in the random sample were then categorised for quality. The year, country of qualification (UK or non-UK) and possession or otherwise of postgraduate qualifications for each of the referring dentists was ascertained from the Dentists' Register. A Standard Normal Deviants (SND), or Z-test was applied to the resulting data. OUTCOME MEASURES: Acceptability or unacceptability of referral letters and proformae, related to the age (assessed by year of qualification), country of qualification and possession or otherwise of postgraduate qualifications. RESULTS: A random sample of 378 referral communications (from a total of 2663) was analysed. Sixty-three of the random sample were referral proformae. The use of the referral proforma was associated with a highly significantly better (P < 0.01) referral performance, than the use of referral letters. There was a trend for referrals from the small numbers of practitioners who were Fellows in Dental Surgery or Masters of Science to be better than those from other practitioners. There was also a trend for use of the referral proforma to enhance the referral performance of practitioners who had been qualified for more than 20 years. Use of the referral proforma highly significantly (P < 0.01) improved the quality of referrals from non-UK qualified practitioners. CONCLUSIONS: In this study, the quality of periodontal referrals was associated for the better with the use of a referral proforma. PMID- 10452191 TI - Integrating outcome measurement with clinical practice: the FACE Recording and Measurement System. Introduction. PMID- 10452192 TI - Process and outcome in mental health care delivery: a model approach to treatment evaluation. AB - In recent years, there has been a strong movement toward "evidence-based medicine," which has significantly affected approaches to the delivery of mental health care. The author highlights the cultural and socioeconomic context in which this trend has emerged. He also notes the resulting controversy over the emphasis on randomized controlled trials (RCTs). Although he does not reject the important role of such trials, he reviews some of their limitations and offers a rationale for the equally important role of naturalistic outcome studies. He concludes by introducing subsequent articles in the journal issue that describe a computerized, online approach for evaluating the effectiveness of mental health care delivery. This system, originally developed in the United Kingdom, has been adapted for use by The Menninger Clinic. PMID- 10452193 TI - The FACE Recording and Measurement System: a scientific approach to person-based information. AB - Modern health care systems increasingly require clinicians to provide person based information for purposes of outcomes and resource management. These demands have traditionally been met by the establishment of parallel data collection activities that can place an excessive burden on both clinicians and the organization. The author describes an approach to information that integrates clinical recording with measurement in a manner that both provides the required information for management purposes and supports day-to-day clinical activity. The Functional Analysis of Care Environments (FACE) Recording and Measurement System integrates clinical, management, and regulatory requirements. In so doing, it also has the potential to provide a basis for effectiveness research and protocol development that can both support and enhance clinical knowledge and practice. PMID- 10452194 TI - Measuring disability and outcomes in routine practice with the FACE Core Assessment. AB - The authors describe the results of an outcome study of 87 psychiatric inpatients and outpatients using a generic assessment and outcome measurement tool, the FACE Core Assessment. The results suggest that the multiaxial structure of the assessment tool has the potential to be used for the routine measurement of clinical outcomes as well as for multidisciplinary assessment. PMID- 10452195 TI - Implementation of an outcomes management model of treatment. AB - The author provides an overview of the new approach to clinical documentation and outcomes assessment and management that is being implemented at The Menninger Clinic as part of a larger effort to systematically study and refine the treatment provided by the Clinic. The scientific, clinical, economic, and regulatory context within which these changes are being made is discussed. Key aspects of the flow of clinical documentation, treatment planning, care recording, and the multifaceted assessment of outcomes and treatment satisfaction are summarized. This article explicates a model for the integration of the processes of clinical information recording with clinical measurement as a means for scientifically evaluating whole clinical systems and their treatment protocols and outcomes. PMID- 10452196 TI - The How Are You? scale: a quality-of-life outcomes measure for routine practice. AB - The How Are You? scale, a quality-of-life outcomes self-report measure to be completed by patients, was developed to address the concerns of the mental health consumer movement. Specific concerns included using less technical language in assessing mental health problems and developing a collaborative dialogue between clinician and patient. The How Are You? scale is a user-friendly instrument that allows patients to be actively involved in their assessment and treatment process. To implement the How Are You? scale into a recording and measurement system, its psychometric properties were assessed using two samples: (1) An employee group from The Menninger Clinic was used to assess internal consistency and stability, and (2) a patient group from The Menninger Clinic was used to assess known-group and discriminant validity, as well as sensitivity to change. The How Are You? scale was found to have good internal consistency and stability and good known-group and discriminant validity. It was also found to be sensitive to change. Although data continue to be collected at The Menninger Clinic, initial indications are that the How Are You? scale is a psychometrically sound outcomes measurement tool. PMID- 10452197 TI - Which treatments help? The patient's perspective. AB - In an effort to determine which specific aspects of treatment might account for patient satisfaction, the authors developed the Components of Treatment Questionnaire. This measure, which was administered to 500 adult inpatients along with the Client Satisfaction Questionnaire (CSQ-8; Larsen, Attkisson, Hargreaves, and Nguyen, 1979), asked patients to assess the helpfulness of their treatment. Convinced that patient input is pertinent to program design, the authors later surveyed 46 patients on treatments that they perceived as helpful and would like to receive. Respondents identified individual relationships and problem-focused therapies as the most helpful aspects of treatment. PMID- 10452198 TI - A simple method of evaluating patients' perceptions of their treatment and care. AB - Surveys of patient satisfaction have become commonplace in mental health services. However, questions about the appropriateness of "satisfaction" as a concept and its use as an approach to evaluating the quality of service provision remain as pertinent as ever. Although surveys of patients of mental health services routinely generate high levels of apparent satisfaction, noncompliance with treatment and medication continues to be a major difficulty in psychiatric care. To evaluate services properly, there is a need to access patients' dissatisfaction to determine whether it is a more valid indicator of quality of services and a better predictor of noncompliance. However, a method that is not time consuming and costly is also required. The authors introduce a new patient evaluation tool, "Your Treatment and Care," which requires patients to report on their direct experience of care according to a set of principles of good practice. The tool forms part of a broader package for obtaining patients' views of their own problems, need for care, quality of care received, and health and social outcomes. The authors also report the results of a survey using this tool. Responses of 75 patients of a psychiatric hospital in the United States are presented and compared with results from a similar survey in the United Kingdom. The comparisons show striking differences in patients' perceptions of quality of treatment and care, both within each population and across clinical contexts. PMID- 10452199 TI - Introduction to the Parent-Child Interaction Assessment. AB - The Parent-Child Interaction Assessment (PCIA) is a procedure developed to yield data about parental attunement as well as other aspects of parent, child, and dyadic functioning. The procedure involves videotaping parents and children ages 4-10 years as they go on an imaginary trip to the zoo together. They begin with a short free-play interaction and then "play out" several scenarios with toy people, animals, and blocks. After cleaning up the toys, the parent and child each are shown the videotape of their interaction. The parent and the child are interviewed about what he or she and the other person are doing, thinking, feeling, and wanting. This article describes the development and theoretical basis of the PCIA. A coding system under development and potential research and clinical applications are discussed. Case examples are provided to illustrate the PCIA. PMID- 10452200 TI - A self-report measure to screen for trauma history and its application to women in inpatient treatment for trauma-related disorders. AB - The authors developed the Trauma History Screen to screen for potentially traumatic events in women admitted for specialized treatment of trauma-related disorders. The questionnaire contains 14 items assessing commonly occurring traumas, and respondents indicate the frequency with which they have experienced the traumas as well as the age at which the trauma occurred. Six items pertaining to interpersonal trauma form an internally consistent subscale. The authors present normative data for a sample of 102 traumatized patients as well as correlations with scores from the Childhood Trauma Questionnaire, the Impact of Event Scale, and the Dissociative Experiences Scale. The findings provide some indication of convergent validity for the Trauma History Screen, and the authors recommend using the questionnaire to alert clinicians to trauma that should be explored more fully in the context of a clinical relationship. PMID- 10452201 TI - Nursing, morality, and emotions: phase I and phase II clinical trials and patients with cancer. AB - This article reflects three nurses' views of the moral dimensions of their work in caring for patients receiving phases I and II of cancer clinical trials in a dedicated cancer clinical trials unit (CCTU). The nurses took part in a semistructured, tape-recorded, group interview in which they talked about any aspect of their work that they felt demonstrated its ethical or moral dimensions. The nurses were not employed as research nurses, but had chosen to specialize in cancer and palliative care in a CCTU environment. Three key themes emerged from the interview: being valued and moral distress; caring in a climate of scientific research; and care, cure, and consequences for moral reasoning. Working in an environment suffused with moral conflicts can be painful and damaging for the professionals involved. It would appear that if nurses are to function effectively, they need to be proactive in promoting an exploration of the role that emotions play in moral decision making and in examining the contribution of emotions to what they care about and why. A commitment to a shared understanding and valuing of divergent ethical reasoning in and across professional cultures of care and research paradigms also appears to be necessary. The terms "ethics" and "morals" are used interchangeably throughout this article. PMID- 10452202 TI - Stress associated with tasks for family caregivers of patients with cancer in Hong Kong. AB - Family caregivers share the strain of cancer and are at risk of physical and psychological symptoms in relation to caregiver stress. This study investigated the relationships between perceived difficulty in managing caregiver tasks and the experience of stress symptoms among 26 family caregivers of terminally ill patients with cancer in Hong Kong. The findings revealed that more stress symptoms were experienced by caregivers who had reported increased difficulty in managing caregiver tasks (rs = 0.64; p < 0.001). Wilcoxon signed-ranks test indicated that psychological stress symptoms, rather than physical symptoms, overwhelmed most of the caregivers (z = -2.15; p < 0.05). There were significantly more negative responses to stress by caregivers who had a shorter duration of experience in providing care. In addition, caregivers who had less education were at higher risk of developing stress (p < 0.01). These findings indicate the need for the provision of support to facilitate the emotional and physical adaptation of caregivers at risk for health problems. PMID- 10452203 TI - Temporal differences in coping, mood, and stress with chemotherapy. AB - This longitudinal study examined relations among mood, coping, perceived stress, and side effects from chemotherapy in 50 individuals with stages III and IV adenocarcinoma of the lung over four consecutive combination chemotherapy courses. Results indicated that perceived stress was moderately high only at the time of pretreatment, and four coping strategies were used: seeking social support, planful problem solving, self-control, and positive reappraisal. No relations existed between coping strategies and side effects from chemotherapy, coping and perceived stress, mood and side effects, and perceived stress and side effects. Seven side effects occurred: leukopenia, decreased activity, nausea, loss of appetite, fatigue, constipation, and taste changes. In summary, receiving chemotherapy is stressful at the time of pretreatment, so nursing interventions need to be concentrated at that point. PMID- 10452205 TI - New agents in gastrointestinal malignancies: Part 2: Gemcitabine in clinical practice. AB - This is part two of the two-part series on new chemotherapy agents for gastrointestinal malignancies. The first article addressed the agent irinotecan hydrochloride (CAMPTOSAR Injection, Pharmacia & Upjohn Company, Kalamazoo, MI; also investigated under the name CPT-11) for use with metastatic colon cancer. This article discusses gemcitabine hydrochloride (GEMZAR for Injection, Eli Lilly and Company, Indianapolis, IN; also referred to as dFdC) for advanced pancreatic cancer. The article reviews the current clinical use, safety profile, and key patient management guidelines for the new and novel cytotoxic agent, gemcitabine, which was approved in 1996. This agent has demonstrated activity in pancreatic cancer, is generally well tolerated, and can safely be administered on an outpatient basis. Therapy-related side effects are manageable with appropriate monitoring and intervention, and reversible with dose modification or discontinuation. As clinical and investigational use of gemcitabine increases, the oncology nurse and other members of the health care team will need to anticipate potential treatment-associated toxicities and be knowledgeable in their early identification and management. As patient advocates, oncology nurses play a key role in treatment outcome and related quality of life through expert patient education, symptom recognition, and intervention according to individual patient tolerance. PMID- 10452204 TI - Comparing patient, parent, and staff descriptions of fatigue in pediatric oncology patients. AB - Fatigue in children and adolescents with cancer is a disruptive symptom meriting clinical intervention by nurses, but has eluded definition, measurement, and intervention. Fatigue in these patients exists within a greater context of illness, treatment, and child and family development. Any effort to define, measure, and intervene with fatigue needs to take into consideration the major components of these children and adolescents' treatment context. The purpose of this descriptive study was to learn from three perspectives (patient, parent, and staff) how fatigue is identified and defined in 7- to 12-year-old children and in 13- to 18-year-old adolescents with cancer, and what factors contribute to or alleviate this fatigue. To elicit this information, separate focus groups were conducted with patients, parents, and staff at two pediatric oncology centers. Resulting interview data were analyzed using pragmatic and semantic content analysis techniques and the Wilson concept analysis process. Findings clearly indicated that children, adolescents, parents, and staff define patient fatigue differently. The conceptual definition from the child data emphasizes the physical sensation of the fatigue; alternating and at times merging physical and mental tiredness are emphasized in the adolescent's definition. Parents and staff view themselves as responsible for alleviating patient fatigue; patients viewed rest and distraction as their primary sources of improving fatigue. The conceptual definitions of fatigue as rendered by our three groups of participants can assist pediatric oncology patients, their parents, and staff in identifying fatigue. Factors identified by these participants as contributing or alleviating fatigue could be the basis for future interventions designed to reduce fatigue in pediatric oncology patients. PMID- 10452206 TI - Cervical cancer screening knowledge and practices among Korean-American women. AB - Cervical cancer is one of the most common cancers of American women. The Papanicolaou (Pap) smear test for cervical screening is a widely used and effective means to reduce the morbidity and mortality rate from cervical cancer through early detection. Despite these benefits, many women have never been screened or are not screened at regular intervals. The purpose of this study was to examine cervical cancer screening knowledge and practices of Korean-American women. The sample consisted of 159 Korean-American women, 40 to 69 years of age. The 1987 Cancer Control Supplement questionnaire was translated into Korean and used to collect data. Twenty-six percent of the respondents never heard of the Pap smear test. Only 34% of respondents reported having had a Pap smear test for screening. The most frequently cited reason for not having had a Pap smear test was absence of disease symptoms. Results indicate that education and usual sources of health care were significant factors related to having heard of or having had a Pap smear test. The findings from this study have important implications for health practitioners and policy makers who serve this ethnic population. PMID- 10452207 TI - Psychosocial determinants of well-being in gynecologic cancer patients. AB - The purpose of this study was to investigate the well-being of women with endometrial versus cervical cancer before treatment, and its determinants at 12 to 15 months follow-up. The subjects were interviewed before treatment, then 6 and 12 to 15 months after initiation of treatment. Simultaneously, they completed questionnaires on psychological symptoms and social support. It appeared that well-being at baseline and at the follow-ups were highly correlated. Furthermore, cervical cancer patients reported significantly more symptoms on all occasions. After discriminant function analysis, the following three variables remained in the analysis: (a) well-being before treatment, (b) diagnosis, and (c) children at home. This allowed 81% of the patients to be classified as having a favorable versus an unfavorable outcome. The results of this study highlight the importance of well-being before treatment as a predictor of well-being after treatment. The necessity of individualized intervention measures is warranted. PMID- 10452208 TI - Transcutaneous electrical nerve stimulation as an adjunct for controlling chemotherapy-induced nausea and vomiting in gynecologic oncology patients. AB - BACKGROUND: To evaluate the efficacy of a miniaturized portable transcutaneous electrical nerve stimulation (TENS) unit (ReliefBand) as an adjunct to standard antiemetic therapy for controlling nausea and vomiting induced by cisplatin-based chemotherapy in gynecologic oncology patients. METHODS: Forty-two patients were enrolled in a randomized, double-blind, placebo-controlled parallel-subjects trial with a follow-up crossover trial. All patients received a standardized antiemetic protocol, then wore the ReliefBand continuously for 7 days. RESULTS: Thirty-two patients were evaluable for the parallel-subjects component, 16 in each group. The percentage of patients with absent or minimal nausea was 59% overall, which was similar to that for both the active (56%) and placebo (62%) groups. The incidence and severity of nausea and vomiting was similar for each group. Eighteen patients completed two consecutive cycles and were evaluable for the crossover component. The average age of the crossover patients and their dose intensity were comparable with those of the overall study population (56.3 versus 58.6 years and 22.7 versus 22.7 mg/m2/week, respectively). The percentage of cycles with absent or minimal nausea was 47% overall, which was similar to that of the active (50%) and placebo (44%) cycles. However, the severity of nausea was significantly lower in the active cycles during days 2 to 4. Patients averaged less than one episode of vomiting daily in each cycle. CONCLUSIONS: The ReliefBand is an effective adjunct to standard antiemetic agents for controlling nausea induced by cisplatin-based chemotherapy in gynecologic oncology patients. PMID- 10452209 TI - An intervention to increase use and effectiveness of self-care measures for breast cancer chemotherapy patients. AB - Outpatient chemotherapy treatment shifts the managing of side effects from health care providers to patients and families. The primary purpose of this quasi experimental study was to determine whether women who receive an intervention of three telephone calls and oral and written self-care measures for specific side effects will use more self-care measures and have higher effectiveness scores as measured by the Exercise of Self-Care Questionnaire (ESCQ) after treatment 4 than women who receive standard care. A second purpose was to determine if women scoring higher on the Exercise of Self-Care Agency (ESCA) Scale before chemotherapy used more and effective self-care measures during chemotherapy than women scoring lower on this scale. Forty-eight women beginning chemotherapy for breast cancer completed the ESCA pretreatment and were interviewed using the ESCQ during a sequence of therapy. The experimental group (n = 26) received additional self-care measures. Subjects with higher ESCA scores before chemotherapy treatment used more self-care measures to alleviate chemotherapy side effects. A telephone call and written self-care measures after the second, third, and fourth chemotherapy treatments did not increase use or effectiveness of self-care measures. The implications of assessing patients' self-care agency before chemotherapy are discussed and a nursing intervention recommended. PMID- 10452210 TI - Breast self-examination training: a brief review. AB - A review of the literature on breast self-examination training indicated that any training in breast self-examination improves compliance, confidence, and proficiency; the evidence is unclear about the relative effectiveness of group or individual training; practice on breast models and on the woman's own breasts should be included in breast self-examination training; additional training sessions improve compliance and proficiency; reminders increase compliance, but the effect ceases when the reminders cease; and it is particularly important for older women to search their breasts slowly and thoroughly. Several other new approaches to breast self-examination training are discussed. PMID- 10452211 TI - Motor coordination in a multi-muscle system as revealed by principal components analysis of electromyographic variation. AB - The variation in electromyographic output of twelve trigeminal muscles of the rabbit was studied to test the hypothesis that they are under the control of a small number of independent neural factors. Jaw muscle electromyograms (EMGs) of eight animals were recorded in 95 chewing sequences, each consisting of 40-75 chewing cycles. The within-sequence correlations of the EMG burst amplitudes (integrated per cycle) and burst onsets were calculated between the muscles. The correlation matrix was subjected to a principal components analysis. This method aims at describing the variation in EMG amplitude and timing by means of the smallest possible set of newly defined variables, or principal components. Of the variation in EMG amplitude values of the twelve muscles, 75-90% could be accounted for by only three principal components. Each principal component was characterized by a group of muscles with high mutual positive correlations; they had zero correlation with other principal components. The first component represents the jaw closers: most of the bilateral masseter and the medial pterygoid muscles. The second represents the openers: the bilateral digastric and lateral pterygoid muscles. This demonstrates the tight control of both the jaw openers and closers, each by a single neural factor; these two factors are independent of one another. They most likely originate from the specific inputs from primary afferents to the opener and closer motoneurons. Unexpectedly, a third independent principal component appeared to control the closing activity of the non-chewing side, posterior deep masseter muscle. It was hypothesized that this muscle acts independently of the other closers to disengage the teeth and resets the jaw for a new chewing cycle. Principal components analysis of variation in timing of EMG onset revealed a grouping of all masticatory muscles in a single cluster, independent of EMG amplitude. This supports the hypothesis that timing and amplitude of masticatory EMG patterns are controlled independently. PMID- 10452212 TI - Action potentials and relations to the theta rhythm of medial septal neurons in vivo. AB - The influence of the medial septal nucleus and the nucleus of the diagonal band of Broca (MS-DB) on the hippocampal theta rhythm includes both cholinergic and gamma-aminobutyric acid (GABAergic) components. To understand the intrinsic septal interactions and the separate contributions of the cholinergic and GABAergic septohippocampal neurons to the theta rhythm in behaving animals, it is essential to be able to identify these two classes from extracellular recordings. Here the durations of extracellularly recorded action potentials are compared with the other characteristics of the neurons. Extracellular recordings were taken from neurons of the MS-DB both in freely moving rats (114 cells) and in urethane-anesthetized rats (112 cells). These were compared with intracellular recordings taken from MS-DB neurons in urethane-anesthetized rats (58 cells). Hippocampal EEG was recorded from above the CA1 pyramidal cell layer (CAI theta) and near the hippocampal fissure (dentate theta) to compare the firing phase across cells. Here it is shown that two major types of rhythmically bursting cells in the MS-DB that had been distinguished previously in intracellular recordings in vivo are also separable in extracellular recordings in vivo on the basis of the durations of their action potentials. In both awake and anesthetized rats the main properties of the two cell types were found to differ: firing rate, phase-relation to the hippocampal theta rhythm and sensitivity of their rhythmicity to blockade of muscarinic transmission. As was previously shown for intracellular recordings in anesthetized rats, it is shown here that in awake rats, too, the more rapidly firing brief-spike (putative GABAergic) cells fired with highest probability on the negative phase of the dentate theta, whereas the more slowly firing long-spike (putative cholinergic) cells fired mostly on the positive phase. Previous work showed that in intracellular recordings from anesthetized rats the rhythmic firing of most brief-spike cells was still retained even during muscarinic blockade, but that of most long-spike cells was lost. Here we also report a recategorization according to spike duration of existing extracellular recordings taken from anesthetized rats, confirming the above observation with much larger numbers of cells. Three additional major new findings are also reported here. (1) In awake rats, muscarinic blockade has relatively little effect on either cell type. (2) Under anesthesia, the firing rates of both cell types are lower than in awake rats, but the effect is greater on the long-spike cells, where the anesthesia also reduces the rhythmicity of the cell firing. (3) Rhythmicity of the putative GABAergic cells is also retained after local injection of GABA-A antagonist, whereas that of the putative cholinergic cells is eliminated. We conclude that either systemic muscarinic blockade or urethane anesthesia alone have relatively little effect on neurons in the defined above MS-DB, but a combination of the two has profound effects on the rhythmicity of the cholinergic cells, largely sparing the GABA-ergic cells. Taken together, the results suggest that generation of theta rhythm requires a background of excitatory influences on the hippocampus (that can be maintained by either muscarinic or glutamatergic inputs) in combination with the phasic disinhibitory action mediated by the GABAergic MS-DB projection. They also provide additional support for the notion that the phasic activity in local collaterals of GABAergic MS-DB cells contributes to the phasic modulation of the firing of cholinergic septohippocampal neurons. PMID- 10452213 TI - Proximodistal structure of early reaching in human infants. AB - Nine infants were tested, at the age of onset of reaching, seated on their parent's lap and reaching for a small plastic toy. Kinematic analysis revealed that infants largely used shoulder and torso rotation to move their hands to the toy. Many changes in hand direction were observed during reaching, with later direction changes correcting for earlier directional errors. Approximately half of the infants started many reaches by bringing their hands backward or upward to a starting location that was similar across reaches. Individual infants often achieved highly similar peak speeds across their reaches. These results support the hypothesis that infants reduce the complexity of movement by using a limited number of degrees-of-freedom, which could simplify and accelerate the learning process. The proximodistal direction of maturation of the neural and muscular systems appears to restrict arm and hand movement in a way that simplifies learning to reach. PMID- 10452214 TI - Dopamine neurons in a simple GDNF-treated meso-striatal organotypic co-culture model. AB - Neurodegeneration of dopamine neurons in the ventral mesencephalon projecting to the dorsal striatum (meso-striatal system) plays a major role in Parkinson's disease. The aim of this study was to establish a simple organotypic, in vitro co culture model for investigating the survival of dopamine neurons stimulated by the novel growth factor, glial-cell-line-derived neurotrophic factor. This model should allow investigation of the effects of the dopaminergic neurotoxin, 6 hydroxydopamine, on the expression of the transcription factor c-fos and on TUNEL staining in vitro. The dopaminotrophic factor, glial-cell-line-derived neurotrophic factor, markedly enhanced dopamine tissue levels and dopamine neuron number. Nerve-fiber ingrowth of dopamine neurons into its striatal target was found to be enhanced with glial-cell-line-derived neurotrophic factor. Using an optimized protocol, it was shown that the neurotoxin 6-hydroxydopamine selectively destructed dopamine neurons. C-fos-like immunoreactivity was enhanced in the mesencephalic part of the co-slices 3 h after application of the neurotoxin. The TUNEL staining occurred 2-5 days after the application of the neurotoxin, but did not seem to be related to dopamine neurons. In conclusion, the organotypic co-culture model provides a simple model for studying survival of dopamine neurons and for observing expression of genes and proteins that could be related to Parkinson's disease. This simple model is useful for screening novel drugs and growth factors and may markedly reduce severe animal experiments. PMID- 10452215 TI - Facilitation of long-term potentiation in layer II/III horizontal connections of rat motor cortex following layer I stimulation: route of effect and cholinergic contributions. AB - The ability of layer I activation to facilitate the induction of long-term potentiation (LTP) in layer II/III horizontal connections of motor cortex (MI) was examined in rat brain slice preparations. Field potentials evoked in layer I and layer II/III horizontal pathways were recorded from radially aligned MI sites. While theta burst stimulation (TBS) of layer II/III pathways alone failed to induce LTP, simultaneous TBS of layer I and layer II/III inputs on alternate sides of the recording electrodes induced LTP in the layer II/III input in 8 out of 13 slices (mean change +20+/-6%; N=13). In the same cases, the layer I connections showed mixed effects: LTP in three cases, LTD in five cases, and no modification in five slices. Despite the facilitatory effect of layer I activation on layer II/III LTP induction, we found that the critical circuitry for this effect was outside layer I. Cutting the layer I fibers selectively in the slice did not prevent layer II/III LTP induction, while cuts preserving only layer I blocked layer II/III LTP after conjoint I+II/III TBS. Cholinergic fibers were evaluated as candidates for the facilitatory effect because they branch widely in both layers and they are thought to participate in synaptic modification. The cholinergic contribution to layer II/III LTP facilitation was investigated using bath application of muscarinic antagonists. Muscarinic blockade prevented facilitation of layer II/III LTP by layer I coactivation. Instead, conjoint stimulation in 10 microM atropine produced long-term depression (LTD) of layer II/III (-18+/-9%; N=11) as well as of layer I (-21+/-6%; N=11) horizontal responses. These results indicate that connections formed within layer I are ineffective in promoting LTP in the deeper-lying horizontal connections; the critical route by which layer I stimulation influenced LTP induction required the circuitry in the deeper layers, particularly the cholinergic system. Thus, it appears that diffuse cholinergic afferents provide an additional route to regulate activity-dependent synaptic modificaton in horizontal cortical connections. PMID- 10452216 TI - Event-related potentials and saccadic reaction times: effects of fixation point offset or change. AB - Previous studies have shown that saccadic reaction times (SRTs) are reduced if the initial fixation point (FP) disappears 200 ms (gap period) before a peripheral target is presented. This gap saccade task is associated with a negative cortical potential at the end of the gap period. To determine whether the neural processes underlying this potential account for the reduction of SRTs during gap saccade tasks, we recorded event-related potentials (ERPs) in 19 subjects performing a gap saccade task (gap duration 200 ms), a warning saccade task (the color of the FP changed 200 ms prior to target appearance) and an overlap task (the FP remained visible during the trial). SRTs were shortest during the gap task, longest during the overlap task and intermediate during the warning task. The gap and warning tasks were accompanied by the same widespread negative cortical potential with a maximum at the time of stimulus presentation. These findings indicate that the warning effect mediated by the disappearance of the FP during gap saccade tasks is responsible for the gap negativity which was observed by several authors. Our findings of shorter SRTs during the gap task than the warning task, however, suggest that the gap has an additional effect that probably depends on subcortical mechanisms. PMID- 10452217 TI - The influence of changes in the stimulation pattern on force and fusion in motor units of the rat medial gastrocnemius muscle. AB - The effects of irregularity in the pattern of stimulation on the tension produced by motor units in the rat medial gastrocnemius muscle were investigated. The effects of decreasing as well as increasing the interpulse intervals were observed for each motor unit in tetani fused to different degrees. For each motor unit type, it was found that the effects of these changes depended on the extent of tetanic fusion. Decreasing the interpulse interval produced an increase in tension during the tetanus: the more fused the profile of tetanus, the smaller the tension increase. Increasing the interpulse interval resulted in a decrease in tetanic tension. This effect was most prominent when the tetanic fusion index was approximately 0.75. This phenomenon resulted from the prolongation in relaxation when tetanic fusion increased, thereby preventing a decrease in tension when the interpulse interval increased. We also investigated the effects of introducing a short interpulse interval ("doublet") at the beginning of the stimulation. The doublets produced increased tetanic tension with a more fused profile. However, the doublet enhanced the sensitivity of the tetanus to increases in interpulse interval and decreased its sensitivity to decreases in interpulse intervals. Slow-twitch motor units appeared to be significantly less sensitive to both increases and decreases in interpulse interval than fast-twitch units. This suggests that slow-twitch units are better suited for producing long lasting contractions with a constant tension level. Conversely, the high sensibility of fast-twitch units to changes in stimulation frequency enhances their participation in regulation of tension of the muscular contraction. PMID- 10452218 TI - Modulation of mRNA expression of the neurotrophins of the nerve-growth-factor family and their receptors in the septum and hippocampus of rats after transient postnatal thyroxine treatment. II. Effects on p75 and trk receptor expression. AB - Early postnatal application of thyroid hormones to rats results in morphological changes of the septo-hippocampal cholinergic and the hippocampal mossy fiber systems. Modulation in the expression of either neurotrophins and/or their receptors is postulated to be involved in these effects. In a recent study, we showed that, after thyroxine application, the mRNA expression of neurotrophins of the nerve-growth-factor (NGF) family is significantly upregulated both in septum and hippocampus. To test whether the neurotrophin receptors (the low-affinity neurotrophin receptor p75 and the specific high-affinity receptors trkA, trkB, and trkC) were also affected by hormone administration, newborn rats were treated daily with subcutaneous injections of thyroxine until postnatal day 12 (P12) at latest. Control animals received corresponding injections of saline. The pups were sacrificed at defined intervals from P9 to P14. The septal areas and the hippocampi were analyzed using the reverse-transcription polymerase chain reaction (RT-PCR) method for quantification of p75, trkA, trkB, and trkC mRNA levels. Analysis of variance over the total investigation period revealed no significant general increases of the gene expressions of either neurotrophin receptor, neither in the septum nor in the hippocampus, although previous results have shown marked changes in neurotrophin levels. On particular postnatal days, significant upregulation could be observed in hippocampus for trkB and trkC. From these and recent data, we conclude that modulation of neurotrophin expression rather than neurotrophin-receptor expression contributes to the morphological modifications within the hippocampal mossy fiber system and the septo-hippocampal cholinergic system. PMID- 10452219 TI - Organization of projections to temporal cortex originating in the thalamic posterior intralaminar nucleus of the rat. AB - Thalamic nuclei surrounding the medial geniculate body, among which the posterior intralaminar nucleus (PIN) is one of the largest, have great importance in fear potentiated emotional behavior. Due to limited knowledge of the distribution of the cortical projections of the PIN, the connections between the temporal neocortex and the PIN were investigated by means of axonal transport of Phaseolus vulgaris leucoagglutinin or Mini-ruby. After iontophoretic injections of either tracer, anterogradely labeled terminals showed a broad, but not a diffuse, distribution in temporal and adjacent cortices (perirhinal, secondary auditory, visceral, secondary somatosensory, agranular insular cortices). A common projection to all areas was found in the upper layer I except for perirhinal cortex, where this projection was confined to the basal layer I. In selected cortical fields (ectorhinal, perirhinal, visceral cortices), an additional projection to layers III/IV was found. The corticofugal projection to the PIN originated from pyramidal neurons in layer V and - in some regions - in layer VI. The present results demonstrate a distinct and selective projection of the PIN to several areas of the temporal neocortex, which may activate inter- and intra areal cortical circuits during processing of auditory stimuli. PMID- 10452220 TI - Effects of GABA-transaminase inhibition on brain metabolism and amino-acid compartmentation: an in vivo study by 2D 1H-NMR spectroscopy coupled with microdialysis. AB - The aim of this work was to study the neurochemical effects in the brain of GABA transaminase inhibition by systemic administration of gabaculine (100 mg/kg, i.a.) in the rat. In order to investigate neurotransmitter and related amino-acid compartmentation and metabolism, we have developed an original tool: the coupling, in vivo, on the same animal, of 2D COSY 1H-NMR spectroscopy with intracerebral microdialysis. The main result is a continuous increase in GABA levels, both in the intracellular compartment (up to 3000+/-450%; P<0.001) and extracellular compartment (up to 808+/-82%; P<0.01) at the sixth hour. The intracellular increase in GABA level became significant at the first hour following gabaculine administration, whereas the extracellular level increased as of the second hour, probably indicating that accumulation of GABA in nerve endings precedes its release in synaptic clefts. Moreover, the levels of the excitatory amino acids, glutamate and aspartate, were decreased both in the intra and extracellular compartments, thus enhancing sedative effects of the drug. We also observed a decrease in the global energetic creatine-phosphocreatine pool, which also could be related to the sedative properties of gabaculine, measurable by the diminution of cortical electrical activity and mean arterial blood pressure. Finally, the coupling between 2D 1H-NMR spectroscopy and intracerebral microdialysis appears to be an original tool for investigating the cerebral metabolic effects induced by pharmacological agents, in situ, in living animals. PMID- 10452221 TI - The effects of blood lactate concentration on perception of effort during graded and steady state treadmill exercise. AB - Studies have reported that ratings of perceived exertion (RPE) are a valid tool for exercise prescription when blood lactate concentration (BLC) is used as the intensity criterion. However, few have studied the relationship between RPE and BLC during commonly used graded exercise tests (GXTs) and simulated exercise training. The purpose of this study was to determine if the RPE: BLC relationship is transferable across GXTs and a steady state exercise trial (SST). Thirteen healthy males (25+/-5.3yrs) completed two maximal treadmill tests (Bruce and Balke protocols) followed by a SST which consisted of approximately 8 minutes of exercise at each of two intensities (approximately 40% and 70% maximal heart rate reserve). BLCs and other physiological measures were compared at matched RPEs across the GXTs and SST trial at each exercise intensity using a two-way repeated measures ANOVA. There were no significant differences in BLC at a matched RPE across the exercise trials at the lower exercise intensity with the BLCs being 1.5+/-0.3, 1.6+/-0.6 and 1.3+/-0.3 mM, respectively. However, at the higher exercise intensity BLCs were significantly lower during the Balke GXT compared to the Bruce GXT and SST (1.8+/-0.6, 2.8+/-1.8 and 3.0+/-0.8 mM, p < 0.05). These results suggest that the RPE: BLC relationship may be protocol dependent during graded exercise testing as it was only transferable from the Bruce GXT to the exercise training setting at intensities in the typical prescription range of 50 85% of VO2max. PMID- 10452222 TI - Direct vs. indirect blood pressure measurement at peak anaerobic exercise. AB - The present study compared the direct intra-arterial method with the indirect conventional sphygmomanometer during all-out anaerobic exercise, in young healthy subjects. Systolic and diastolic blood pressures were measured by intraarterial means and by auscultation. Fourteen young healthy males (23+/-2 yrs) were measured at rest and during all-out anaerobic exercise. Comparisons were made with simultaneously determined intra-arterial catheter and auscultation measurements. The data suggest that indirect systolic pressure is highly correlated with the direct method at rest (r = 0.684), with mean of 107+/-7 and 101+/-6 mmHg, respectively, and during all-out anaerobic exercise (r = 0.87), with mean of 197+/-11 and 191+/-9 mmHg, respectively. Indirect diastolic blood pressure correlates well with intra-arterial at rest (r = 0.62), with mean of 84+/-11 and 77+/-9 mmHg, respectively. However, during all-out anaerobic exercise, the correlation coefficient between the direct and the indirect methods was low (r = 0.36), with mean of 101+/-12 and 103+/-9 mmHg, respectively. These results suggest that when utilizing an all-out anaerobic exercise, the indirect method is not valid for assessment of diastolic pressure. In addition, although the anaerobic test is a dynamic type of exercise, its blood pressure responses for both direct and indirect methods were similar to those seen during isometric exercise. PMID- 10452223 TI - Human muscle energetics during voluntary and electrically induced isometric contractions as measured by 31P NMR spectroscopy. AB - Electrical stimulation (ES) and voluntary contraction (VC) were compared in the quadriceps muscle of ten male volunteers. In both modes, a workload corresponding to 20% of maximal voluntary contraction was applied during 64 isometric contraction (5.5 s)-relaxation (5.5 s) cycles. The protocols were performed in a 1.5 T whole-body magnet. The Pi/PCr ratio and the intracellular pH (pHi) were monitored by 31P NMR spectroscopy during baseline, exercise and recovery periods, in a superficial region of the vastus medialis. During baseline, the Pi/PCr ratio (0.12 vs. 0.10) and the pHi (7.01 vs. 7.00) were comparable in both conditions. During exercise, the Pi/PCr ratio was higher (0.36 vs. 0.14) and the pHi was lower (6.85 vs. 7.07) during ES than during VC. For the same external work production, these results reflect a different metabolic solicitation in the ES quadriceps than in the VC ones. PMID- 10452224 TI - Doubly labelled water validation of three physical activity questionnaires. AB - This study investigated the validity of the Baecke Questionnaire, the Five City Project Questionnaire, and the Tecumseh Community Health Study Questionnaire in 19 Flemish males, using correlation and multiple stepwise regression analyses. The three questionnaires are commonly used physical activity questionnaires in epidemiological studies. The physical activity level (PAL) as measured with the doubly labelled water method was used as the criterion. The Baecke total activity index showed the highest correlation coefficient with PAL (r = 0.69, p < 0.001). Also the sweat index from the Five City Project Questionnaire, and total daily energy expenditure from the Tecumseh Community Health Study Questionnaire showed significant associations with PAL, respectively 0.57 (p < 0.05) and 0.64 (p < 0.01). Multiple stepwise regression analyses supported the findings from the correlation study. The largest individual contribution in PAL was from the activity index (45%) for the Baecke Questionnaire, the sweat index (29%) for the Five City Project Questionnaire, and total daily energy expenditure (38%) for the Tecumseh Community Health Study Questionnaire. In conclusion, the questionnaires, and certainly the Baecke Questionnaire, can provide valid data about physical activity. Therefore they are useful in epidemiological studies. PMID- 10452225 TI - Upper body power comparison between groups of cross-country skiers and runners. AB - The purpose of this study was to evaluate relationships between upper body power (UBP) and cross-country ski skating race velocity (R) in a cross sectional representation of cross-country skiers. Additionally, cross-country skiers' UBP was compared to UBP of distance runners. Participants (n = 195) were tested on a Street Arm Ergometer for UBP using a ramped maximal UBP test simulating a double poling motion. A strong relationship (r = 0.89) between UBP and RV in skiers was determined. High school skiers were separated into slow and fast groups based on reported RV; significant differences (p < 0.05) in both UBP was found to be an effective partial predictor of RV, independent of gender. Mean UBP for the cross country runners was 46% of mean UBP for the cross-country skiers. With UBP contributing so much to performance in cross-country ski racing, it is recommended that cross-country skiers focus a large portion of their training on the specific development of that fitness component. PMID- 10452226 TI - A comparison of drop jump training methods: effects on leg extensor strength qualities and jumping performance. AB - The purpose of this study was to determine the effects of two drop jump (DJ) training methods on the strength qualities of the leg extensors and vertical jumping performance. Thirty-five males were allocated to either a control group or one of two training groups, required to perform 72-90 DJ per week for 6 weeks: DJ for maximum rebound height (DJ-H) or DJ for maximum height and minimum ground contact time (DJ-H/t). The subjects were tested before and after the training on leg extensor function for maximum strength and speed-strength under concentric and stretch-shortening cycle muscular actions, and on vertical jumps from a standing position and a run-up. ANOVA with repeated measures showed that a 20% gain in reactive strength (RS) for the DJ-H/t group was significantly greater (p < 0.05) than the other groups, however, this did not transfer to vertical jumping performance. The DJ-H group did not achieve a significant training effect. It was concluded that DJ-H/t method was effective for the development of RS, but training with DJ-H was not intense and/or specific enough to stimulate gains in strength qualities of the leg extensors or jumping performance. PMID- 10452227 TI - Validation of Aerosport KB1-C portable metabolic system. AB - The purpose of this study was to test the validity of the Aerosport KB1-C portable metabolic measurement system against the criterion Douglas bag method. During cycle ergometry, simultaneous measurements of minute ventilation (VE), oxygen consumption (VO2), and carbon dioxide production (VCO2) were made at rest and at power outputs of 50, 100, 150, 200, and 250 W. No significant differences (P > 0.05) were observed for VE, VO2, and VCO2 between the KB1-C and Douglas bag at 100, 150, and 250 W, while the KB1-C was significantly different (P < 0.05) from Douglas bag values at rest, 50, and 200 W. R values were found to be significantly different (P < 0.05) at 100, 150, and 200 W, while no significant differences were observed at rest, 50, and 250 W. The fractional concentrations of oxygen (FEO2) and carbon dioxide (FECO2) were not significantly different at 50, 100, 200, and 250 W while values at rest and 150 W were significantly different (P < 0.05). These findings show that the Aerosport KB1-C portable metabolic system is acceptable for measuring oxygen uptake in the range of 1.5 and 3.5 L x min(-1), using the medium flow pneumotach setting. At lower intensities, the low-flow pneumotach setting provides acceptable results. PMID- 10452228 TI - Effects of branched-chain amino acids and carbohydrate on fatigue during intermittent, high-intensity running. AB - Experimental support for the hypothesized benefits of BCAA supplements on endurance performance is limited. However, it is theorized that the benefits may be enhanced if 1) BCAA are taken along with a pre-event carbohydrate meal as well as during exercise, and 2) the exercise is intermittent in nature. This study tested the effects of ingesting carbohydrate beverages with and without BCAA before and during intermittent high-intensity running to fatigue. Eight subjects performed 3 exercise trials consisting of intermittent shuttle running (walking, sprinting, and running) to fatigue. Subjects drank either carbohydrate drinks given 1 h before (5 mL/kg, 18% carbohydrate) and during exercise (2 mL/kg, 6% carbohydrate) (CHO), carbohydrate drinks with BCAA (7 g) added to the portions consumed 1 h before and immediately before exercise (CHO+BCAA), or flavored water placebos (P). Subjects ran longer when fed either CHO or CHO+BCAA as compared to P, with no differences between CHO and CHO+BCAA. CHO and CHO+BCAA also had higher plasma glucose and insulin, and lower FFA (p < 0.05). These findings confirm a beneficial effect of carbohydrate feedings on fatigue during exercise designed to mimic the activity pattern that occurs in sports like soccer, basketball, and hockey. They do not, however, support the hypothesis of an added benefit of BCAA supplements. PMID- 10452229 TI - The effect of arginine or glycine supplementation on gastrointestinal function, muscle injury, serum amino acid concentrations and performance during a marathon run. AB - Gastrointestinal bleeding and increased intestinal permeability have been observed in marathon runners. We sought to determine if L-arginine would be useful for prevention of these complications. Twenty-three runners were randomized to receive L-arginine (A) or glycine (placebo) (G), 10 grams 3 times daily for 14 days prior to the 1997 Houston-Methodist Marathon. Serum, stool hemoccults and lactulose:mannitol permeabilities were obtained at baseline, immediately after completion of the marathon and approximately 48 hours later. Runners rated their symptoms of nausea and vomiting, belching and indigestion, abdominal pain and bloating, diarrhea, and extremity pain on a 1-5 scale of increasing severity. The L:M was unchanged in either group during the three collections. Occult bleeding occurred in 8%/20% in A and G groups, respectively, p = NS) immediately post-marathon. No runners had occult bleeding 48 hours post race. Gastrointestinal symptom scores were minimal to nonexistent. Extremity pain scores were similar for groups A and G (2.1+/-1.4 and 2.8+/-1.6, respectively, (p = NS). Fluid intake was similar between both groups (1875+/-1547 vs. 1506+/-970 ml, p = NS). Serum amylase was normal at baseline and remained virtually unchanged. Serum lipase was normal at baseline and immediately post-race in both groups, but increased at 48 hours post-race (82.2+/-34.3 to 121.5+/-53.3 mg/dl [A], p = 0.02 and 114.3+/-55.7 to 181.9+/-162.2 mg/dl [G], p = 0.09). CPK increased significantly and similarly in both groups immediately post-race, and even more dramatically 48 hours post-race (130.3+/-130.8 to 738.8+/-902.9, p = 0.007 to 1966.5+/-3.166.0 mg/dl [A] and 140.9+/-77.9 to 863.0+/-772.3, p = 0.003 to 5619+/-10636.8mg/dl [G]). Modest post-race decreases were seen in most serum amino acids in both groups. Finish times were longer than predicted (23+/-21 and 9+/-7 min for A and G groups, respectively, p = 0.049). Our study failed to show a clear benefit of arginine supplementation for the prevention of intestinal ischemia/reperfusion injury associated with endurance running, but either a detrimental affect on performance with arginine, or enhanced performance with glycine. Skeletal muscle injury was unaffected by arginine or glycine supplementation. The delayed increase in serum lipase suggests mild pancreatic injury, affected by either arginine or glycine supplementation. PMID- 10452230 TI - Blood leukocyte and glutamine fluctuations after eccentric exercise. AB - Skeletal muscle, as a producer of glutamine, is important for lymphocytes, monocytes and macrophages. Exercise-induced muscle damage could burden the immune system by concurrently eliciting a local inflammatory response and decreasing glutamine availability. The aim of this study was to determine whether blood leukocyte and glutamine concentrations were affected in individuals with high serum creatine kinase (CK) activity (indirect indication of muscle damage) compared to those with no change in CK. Twelve females performed maximal eccentric resistance exercise using one arm and one leg. Blood leukocyte subsets and glutamine were measured at 24 and 0 h pre-exercise, and post-exercise at intervals up to 9 d post-exercise. Eleven subjects were placed in High (n = 6) and Low CK (n = 5) groups. Lymphocytes, (total, natural killer, and T), monocytes, and granulocytes did not change significantly in either group, at any time. Whole blood glutamine concentration decreased (p < 0.05) from 437 microM pre-exercise to 332 microM 3 d post-exercise in both groups. The decrease in glutamine suggests that the metabolism of the muscle may be affected by this exercise, however, the occurrence of this decrease in both groups suggests that this change was not a response to muscle damage. PMID- 10452231 TI - Skull bone mass deficit in prepubertal highly-trained gymnast girls. AB - It is known that impact loading sport can increase the bone mineral density in the stressed sites of the skeleton in athletes. However, non weight-bearing sites are seldom studied in healthy young girl athletes. In order to study the effects of a long term intensive training on the non-stressed region of the skeleton (skull), we investigated both highly-trained girl athletes, involved in sports requiring or not significant impact loading on the skeleton and a girl control group. Bone mineral content (BMC) and density (BMD) were measured in the whole body, at lumbar spine, femoral neck, trochanter, Ward's triangle, radius, head and ribs, in 60 prepubertal girls including 12 swimmers, 32 gymnasts and 16 controls. Measurements were made by DXA. There were no statistical differences between the groups as regards age, height, body weight, body mass index, lean tissue mass and dietary calcium intake. Mean BMD in gymnasts was statistically higher than in other groups for radius (p < 0.001), femoral neck (p < 0.05) and Ward's triangle (p < 0.05) while there was no difference between swimmers and controls. Head BMC was significantly lower in gymnasts compared to other groups (241.9+/-41 g vs. 285.8+/-34.7 g and 291.1+/-50.2 g respectively in swimmers and controls, p < 0.001). The same observation was made for head BMD (p < 0.01). When body weight was used as a covariant, the contribution of the head BMC to the whole body was significantly lower (p < 0.001) in gymnasts (24.97%) than in swimmers (27.88%) and controls (27.77%). When compared between groups, the slopes of the regressions for head/whole body BMC or BMD were significantly lower in gymnasts (p < 0.05) than in other groups. These data suggest that in prepubertal children the increased bone density induced by gymnastic training in the stressed sites of the body could be related to a decreased skull bone mass. PMID- 10452232 TI - Parental attitudes about exercise regarding their children with cystic fibrosis. AB - Despite the well-recognized benefits of exercise on general health and pulmonary function, lack of participation in regular exercise remains a concern with respect to children with cystic fibrosis (CF). Data are lacking regarding parental perceptions about exercise in children and adolescents with CF. Sixty nine parents of children with CF and 70 parents of healthy children completed the 'Exercise Benefits/Barrier Scale" self administered inventory. Fifteen additional items addressing exercise issues for people with CF were also included. Data were reported as total score, and subdivided into barriers and benefits scales for analysis. Data on the CF specific questions were also reported. Parents of children with CF scored lower on both the total score and benefits portion of the inventory and scored slightly greater on the barriers portion (p < .05) representing less favorable perception of the benefits of exercise along with increased barriers. The presence of a healthy child in addition to a child with CF appeared to increase both the total score and benefits score as well as resulting in a more positive outlook on exercise related uses in CF. It appears that significant differences in parental perceptions regarding exercise indeed exist not only when compared to healthy children but within varying intra structures of families with a child who has CF. PMID- 10452233 TI - Re: Vehrs, P., Morrow, J.R., Butte, N.: Reliability and concurrent validity of futrex and bioelectrical impedance. Int J Sports Med 19: 560-566, 1998. PMID- 10452234 TI - P-glycoprotein-expressing tumor cells are resistant to anticancer drugs in human gastrointestinal cancer. AB - The resistance to doxorubicin (DOX) by some tumor cells is mainly due to the effect of P-glycoprotein encoded by the multidrug resistance-1 (mdr1) gene. We tried to prove the correlations between P-glycoprotein expression and the sensitivity for anticancer drugs including DOX and other cytotoxic drugs that are currently used for gastrointestinal cancer patients. We quantified the P glycoprotein expression by flow cytometry techniques, and the sensitivity for anticancer drugs using a tetrazolium salt, 3-(4,5-di-methylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT), assay in highly purified fresh human tumor cells obtained from 25 cancer patients. The inhibition rates were the lowest in DOX and mitomycin C (MMC), compared with other drugs. The most significant correlation between DOX and MMC was seen in the inhibition rates. A significant correlation was also seen between the inhibition rates for DOX and P-glycoprotein expression, whereas only a slight correlation between the sensitivity for MMC and P-glycoprotein expression was observed. We should therefore pay close attention to the effect of P-glycoprotein when treating cancer patients, especially if both the inhibition rates of DOX and MMC are low based on the findings of an MTT assay. PMID- 10452235 TI - Functional results of colonic J-pouch anastomosis for rectal cancer. AB - The purpose of this study was to clarify the functional outcomes of colonic J pouch anastomosis (J-LAR) for lower rectal cancer in comparison with those of traditional straight anastomosis (S-LAR). A questionnaire regarding anorectal function was conducted 1 year after operation on patients who underwent J-LAR (n = 15) and S-LAR (n = 30). The clinical functions were assessed by an incontinence scoring system. The physiologic function was assessed by anorectal manometry and the balloon expulsion test. No patients demonstrated a diverting stoma. The bowel frequency (range) 1 year after operation was 4.8 (3-6) in the S-LAR group and significantly decreased to 1.8 (1-3) in the J-LAR group (P < 0.05). Complete evacuation was 50.2% (40%-60%) in the S-LAR group and significantly increased to 80.6% (60%-90%) in the J-LAR group (P < 0.05). Neorectal compliance was 2.2 (1.4 2.9) ml/mmHg in the S-LAR group and significantly increased to 3.1 (1.3-3.5) ml/mmHg in the J-LAR group (P < 0.01). No significant difference was observed between the two groups regarding the maximum resting or maximum voluntary squeezing pressure. In conclusion, our findings suggested colonic pouch anastomosis performed after a low anterior resection to support the compliance of the (neo)rectum to be an important factor for retaining a satisfactory bowel frequency. PMID- 10452236 TI - The effect of thorough preoperative disinfection on the incidence of postoperative mediastinitis. AB - This study was conducted to evaluate the effectiveness of scrubbing the skin of patients scheduled to undergo elective cardiac surgery with disinfectant solutions in an attempt to reduce the incidence of the potentially fatal postoperative complication of mediastinitis. Since April 1991, we have routinely disinfected the skin of all such patients three times preoperatively. A retrospective comparison of the incidence of mediastinitis before and after April 1991 was carried out. Microbiological examinations of the anterior chest were conducted in ten patients admitted for elective operations, and in four patients admitted for emergency surgery. Specimens were obtained before each scrubbing, before the operation, and just after skin closure. There was a significant decrease in the general incidence of mediastinitis after April 1991 (P < 0.01). Microbiologically, significant decreases in colony counts were also observed after the first (P < 0.05) and second scrubbing (P < 0.05) in the ten patients specifically studied before elective surgery. No colonies grew in the postoperative specimens from any of these patients, but colony formations were observed in two of the four patients who underwent emergency surgery (P = 0.066). Thus, we conclude that our thorough disinfection method effectively achieves adequate, prolonged suppression of the skin flora, which explains the marked decrease in the incidence of mediastinitis in our hospital since its institution. PMID- 10452237 TI - Intrathoracic light-assisted anterior limited thoracotomy in lung cancer surgery. AB - We recently developed an intrathoracic light-assisted anterior limited thoracotomy (ILAALT) for use in lung cancer surgery. A skin incision 12cm long is made below the breast, then the pectoral major muscle is divided, and the fourth intercostal space is opened with a disconnection of the anterior cartilagenous portion. The posterior skin, including the serratus anterior muscle, is drawn posteriorly using a retractor. To illuminate the posterior and apex portions of the thoracic cavity, a flexible fiber light is introduced into the thoracic cavity through the eighth intercostal space at the posterior axillary line. These techniques provided adequate exposure and sufficient illumination in the thoracic cavity, thus making surgery easy for most thoracic applications. Using this approach, we undertook 28 lung resections with a mediastinal nodal dissection for lung cancer (24 lobectomies, 2 bilobectomies, and 2 pneumonectomies) without difficulty. The mean intrasurgical blood loss was 217ml, the operative time 262min, and chest tube drainage duration 2.3 days. Except for one case, no patients required a blood transfusion. All patients underwent continuous epidural anesthesia until postoperative day (POD) 8. The mean time that other analgesic medication was required was 0.5 times per patient until POD 13, but none from POD 14 on. We thus conclude ILAALT to be low-invasive thoracotomy and is thus indicated for most types of lung cancer surgery, providing a reduction of pain as its main advantage. PMID- 10452238 TI - How different surgical methods of performing right upper lobectomy contribute to postoperative bronchial branching deformity: an experimental study. AB - It has been suggested that postoperative bronchial deformity is a complication of surgery and that the surgical methods employed may play a role in its development. We studied the relationship between postoperative bronchial branching deformity and various surgical methods, including hand suturing or mechanical stapling of right upper (RU) lobectomy, versus wedge or sleeve bronchoplasty with RU lobectomy. We used an in situ bronchial casting model in rabbits and measured the branching angles between: the trachea and the right main bronchus (RMB); the trachea and the left main bronchus; the RMB and the intermediate bronchus (IB); the IB and the right middle lobe bronchus (RMLB); and the RMLB and the coronal plane. In the mechanical stapling group, the angle between the RMB and the IB was wider than in the hand suturing group, and the RMLB diverged more laterally. In the wedge plasty group, the angle between the trachea and the RMB was wider, while that between the IB and the RMLB was narrower than in the hand suturing group. In the sleeve plasty group, the angle between the RMB and the IB and that between the IB and the RMLB were wider than in the hand suturing group. The findings of this experiment clearly demonstrate that postoperative deformity of the distal bronchus varies according to the surgical methods used. PMID- 10452239 TI - Distribution and localization of cells and collagens in the proliferated intima of arterially implanted autovein grafts. AB - We examined the microscopic features and distribution of collagens in the hyperplastic intima of arterially implanted autovein bypass grafts under conditions of a reduced blood flow with a poor distal outflow. Vascular anastomosis was made using 7-0 nonabsorbable polypropylene sutures (PP group), or absorbable polydioxanone sutures (PDS group). On the contralateral limb, an autovein bypass graft was performed under normal flow conditions (NF group). The thickness of the intima in the NF group was approximately 50 microm throughout the duration of the study, while in the PP and PDS groups, intimal hyperplasia progressed to 290+/-112 microm and 267+/-123 microm, respectively, at 13 months after grafting. Collagen accumulated significantly in both the PP and PDS groups; types IV and V collagen in particular increased considerably in the deep layer. Regardless of the suture materials, the progression of intimal hyperplasia was considered to be closely related to the poor distal outflow to be and caused by the proliferation of myofibroblasts and active production of collagen. The increase in types IV and V collagen, particularly in the deep layer of the hyperplastic intima, was due to development of numerous vasa vasora in this region. PMID- 10452240 TI - Tetanus after a resection for a gangrenous perforated small intestine: report of a case. AB - We report herein the case of a 75-year-old man who developed severe tetanus 24 h after the resection of a gangrenous perforated small intestine. It seemed that the tetanus was caused by a spillage of the intestinal contents harboring Clostridium tetani; however, this was not identified by a culture. The diagnosis of tetanus was made only when opisthotonus in this patient became evident and normal tetanus treatment proved to be successful. PMID- 10452241 TI - Intussusception of a mucocele of the appendix secondary to an obstruction by endometriosis: report of a case. AB - We treated a patient with a complete invagination of the cecum which contained a mucocele of the appendix secondary to an obstruction by endometriosis. Preoperatively, a barium enema showed a crab's claw-like area without filling in the oral side of the transverse colon. An emergency laparotomy was performed and revealed a mucocele of the appendix to have induced appendicecal invagination; however, no colonic invagination was observed. An appendicecal resection was thus done. Pathologically, the resected specimen was a mucosal hyperplasia with mucin secreting lesions of the appendix. The theories regarding the pathogenesis of appendicecal mucocele are reviewed and discussed. PMID- 10452242 TI - Peritoneal recurrence of colon cancer detected by positron emission tomography: report of a case. AB - Increased glucose metabolism has been reported to occur in association with colorectal cancer. As positron emission tomography (PET) using [18F]fluorodeoxyglucose is able to depict hypermetabolic sites, it can therefore be used to detect colorectal cancer. A 69-year-old male patient with a recurrent solitary liver metastasis from colon cancer underwent whole-body PET which revealed high [18F]fluorodeoxyglucose uptake in the lesion. Furthermore, PET revealed peritoneal metastases that had not been detected by conventional imaging methods. Consequently, PET proved useful in helping us to avoid performing unnecessary treatment for the liver metastasis. Although it is uncertain whether early identification of recurrence can prolong survival, it may help to prevent unnecessary treatments being carried out. Thus, the application of PET in carefully selected patients could be beneficial to the management of recurrent colorectal cancer. PMID- 10452243 TI - Diffuse schwannoma involving the entire large bowel with huge extramural development: report of a case. AB - Schwannoma of the large bowel is a rare clinical entity, which has reportedly been recognized to arise from one place with a submucosal tumor morphology. We present herein the unique case of a 25-year-old woman who suffered from a schwannoma diffusely involving the entire large intestine. The patient complained of abdominal distension and imaging studies revealed a giant tumor occupying the whole abdomen, but no confirmed preoperative diagnosis could be made. A laparotomy proved the huge tumor detected preoperatively to be the markedly wall thickened entire large bowel itself due to the diffuse extramural development of a neoplasm, but no other organs were involved. Biopsy specimens from the tumor were histologically diagnosed as benign schwannoma. However, because of the possibility of malignancy, we later performed a total proctocolectomy followed by an ileal J-pouch-anal canal anastomosis. The final pathological diagnosis was also that of a benign schwannoma originating from the large intestine. The patient remains well without recurrence 15 months after surgery. To the best of our knowledge, no such case of a schwannoma in the entire large bowel has yet been reported in the literature, and the pathogenesis of its occurrence remains unknown. PMID- 10452244 TI - A solitary fibrous tumor in the perianal region with a 13-year follow-up: report of a case. AB - A case of a solitary fibrous tumor (SFT) of the perianal region in a 62-year-old man is reported herein. The patient had undergone an abdominoperineal excision of the rectum for an anorectal tumor 13 years previously, and had been referred to our hospital for a perineal mass. Computerized tomography and angiography revealed a markedly hypervascular tumor measuring 11 x 8 cm in size in the pelvic cavity. After preoperative radiotherapy (total 58 Gy) and the embolization of the feeding arteries, he underwent an en bloc excision of the tumor. Microscopically, the tumor was composed of spindle shaped cells with a "patternless" arrangement in a collagenous background. There was immunohistochemical evidence that these cells were strongly positive for CD34, thus suggesting the tumor to be SFT. The previously resected anorectal tumor showed similar histological and immunohistochemical findings. The patient's recovery was uneventful. PMID- 10452245 TI - Right hepatic lobectomy for hepatocellular carcinoma which developed in primary biliary cirrhosis: report of a case. AB - The case of a 74-year-old female patient who underwent a right hepatic lobectomy for hepatocellular carcinoma (HCC) which developed in primary biliary cirrhosis (PBC) is reported herein. During a follow-up examination for Parkinson's disease, an elevation of hepatobiliary tract-related enzymes and alpha-fetoprotein was uncovered. Diagnostic imagings showed a hypervascular, solitary, and encapsulated tumor measuring about 7 cm in diameter located mainly in the posterior segment. Positive antimitochondrial and antinuclear antibodies and a preoperative liver biopsy strongly suggested well differentiated HCC developed in PBC (Scheuer's classification stage II). Since the natural prognosis of PBC estimated by the Mayo risk score was fairly good and the liver function indicated sufficient tolerance for major hepatic resection, and preoperative computed tomography (CT) volumetry showed the atrophy of the right hepatic lobe, a right hepatic lobectomy was performed. A pathological examination revealed well encapsulated, moderately differentiated HCC with, in part, well-differentiated HCC in the tumor and stage II PBC in the noncancerous region. CT volumetry performed at postoperative day 14 showed a 146% enlargement of the remnant liver. An early detection of HCC and PBC by strict screening would prevent a limitation of surgical therapy due to a deteriorated liver function. PMID- 10452246 TI - Double cancer of the gallbladder and common bile duct associated with an anomalous pancreaticobiliary ductal junction without a choledochal cyst: report of a case. AB - We report herein the case of a 37-year-old woman found to have double cancer of the gallbladder and common bile duct associated with an anomalous pancreaticobiliary ductal junction (APBDJ) without a choledochal cyst (CC). Abdominal ultrasonography showed an isoechoic mass in the gallbladder, and percutaneous transhepatic biliary drainage tubography revealed incomplete obstruction in the upper portion of the common bile duct and APBDJ. The patient underwent cholecystectomy, partial hepatic resection, pancreatoduodenectomy, and portal vein reconstruction. Pathological examination of the tumors from the gallbladder and bile duct revealed papillary carcinoma and poorly differentiated adenocarcinoma, respectively, and direct continuity was not observed between the tumors. A review of the literature on six cases of multiple primary carcinoma of the biliary tract associated with APBDJ without CC is presented following this case report. Double cancer of the biliary tract was found synchronously in five patients and metachronously in one. Gallbladder cancer showed subserosal invasion in four patients, while bile duct cancer invaded the pancreas in one patient and reached the serosa in two patients. Considering the potential for cancer to arise in the biliary tract and the difficulties associated with monitoring it, cholecystectomy and resection of the extrahepatic common bile duct may be the most appropriate treatment for patients with an APBDJ without a CC. PMID- 10452247 TI - Ruptured superior mesenteric artery aneurysm occurring in association with a heterotopic pancreas: report of a case. AB - Aneurysmal disease of the visceral arteries is found in only about 0.2% of the population, and the celiac trunk and superior mesenteric artery (SMA) are involved in less than 10% of all visceral aneurysms. We present herein the case of a 71-year-old woman who suffered rupture of a SMA aneurysm. Histological examination of the periarterial tissues which existed next to the aneurysm revealed a heterotopic pancreas. To the best of our knowledge, no other case of an SMA branch aneurysm presenting in association with a heterotopic pancreas has ever been described in either the Japanese or English literature. This is the first report to indicate that a heterotopic pancreas is a likely incidental factor predisposing to visceral aneurysms. PMID- 10452248 TI - Dermoid cyst of the spleen: report of a case. AB - A rare splenic dermoid cyst in an 11-year-old girl is reported herein. The patient was referred to us with the chief complaint of acute-onset abdominal pain. The lesion was demonstrated by ultrasonography, computed tomography, and magnetic resonance imaging to be a cystic lesion containing serous fluid. A splenectomy was performed. Both histopathological and immunohistochemical findings demonstrated the cyst to be a dermoid cyst of the spleen. To avoid the risk of infection following splenectomy, polyvalent pneumococcus vaccine was administered. The patient recovered uneventfully. We performed immunohistochemical studies with cytokeratin and electron microscopy to determine the origin of the cyst. PMID- 10452249 TI - Epiphrenic diverticulum composed of airway components attributed to a bronchopulmonary-foregut malformation: report of a case. AB - Bronchopulmonary-foregut malformation (BPFM), defined originally as pulmonary sequestration with or without communication to the esophagus, has been acknowledged to include congenital foregut diverticula. We present herein the case of a 43-year-old woman with a 9-year history of dysphagia, in whom a barium meal examination demonstrated a 2.5-cm epiphrenic diverticulum and several fistulae. A laparotomy was performed and the lower esophagus without communication to the lung was pulled down and resected, followed by an esophagogastrostomy carried out with fundopexy. Since her operation, the patient has been free of symptoms. Histologically, the diverticulum was observed to be lined by stratified squamous cells, but its shape was formed by mural cartilage, smooth muscle cells, and three ciliated-cell cysts. The dysphagia was considered to have been derived from the kinked esophagus created by the rigid diverticulum, being the possible developmental arrest of a supernumerary lung bud. These findings indicate that this case may involve BPFM in the broad sense. Although several cases of bronchogenic cysts located beneath or across the diaphragm have been reported as a subgroup of BPFM, congenital epiphrenic diverticula has rarely been described. PMID- 10452250 TI - The marked anticancer effect of combined VCR, MTX, and indomethacin against drug resistant recurrent small cell lung carcinoma after conventional chemotherapy: report of a case. AB - In a previous study, we discovered that indomethacin was an effective modulator of the sensitivity of pulmonary carcinoma cells to vincristine (VCR), methotrexate (MTX), adriamycin (ADR), and etoposide (VP-16). We describe herein the case of a 61-year-old-man with multiple brain, lung, liver, and bone metastases from small cell lung carcinoma (SCLC) that recurred after intensive chemotherapy, who showed no signs of remission following conventional chemotherapy. The general condition of the patient deteriorated until he required morphine sulfate to control his severe diffuse pain. In an attempt to improve this patient's quality of life (QOL), he was discharged from hospital and treated at the outpatient clinic with modulation therapy using indomethacin as an anodyne instead of morphine sulfate. Signs of almost complete remission to only one cycle of combination therapy with VCR, MTX, and indomethacin were observed without any obvious adverse effects. This case report serves to demonstrate that modulation therapy combined with VCR, MTX, and indomethacin may be useful in the treatment of patients with drug-resistant recurrent SCLC. PMID- 10452251 TI - Mediastinal lymph node involvement as the initial manifestation of occult thyroid cancer in the surgical treatment of lung cancer: report of a case. AB - A 63-year-old man was referred to our institute for the treatment of squamous cell carcinoma of the upper lobe of his right lung. A right upper lobectomy of the lung was performed with a mediastinal lymph node dissection. The postoperative pathological examination of the dissected specimens revealed one of the superior mediastinal lymph nodes to be morbid with micrometastasis of occult thyroid cancer, while no node involvement was seen due to lung cancer. A right lobectomy of the thyroid gland with a modified radical neck dissection was done 4 years later after the confirmation of the absence of any recurrent sign of lung cancer. In the resected specimen, papillary thyroid microcarcinoma was observed with several intraglandular metastases and right regional lymph node involvement. Eight months later, a new primary lung cancer developed in the left lung, and a left upper lobectomy of the lung with a mediastinal lymph node dissection was performed. At that time, the absence of mediastinal lymph node metastasis from lung cancer or thyroid cancer was confirmed. Mediastinal lymph node involvement as the initial manifestation of occult thyroid cancer in surgical treatment for lung cancer is rare, but it is important to be aware of the possibility of incidentally detecting occult thyroid cancer in surgical dissections in this area for lung cancer. The appropriate surgical treatment should be determined while carefully considering the prognosis of the lung cancer as well as that of any coexisting malignancy. PMID- 10452252 TI - An aberrant left subclavian artery aneurysm with right aortic arch: report of a case. AB - The case of a 41-year-old man who developed an aneurysm in his aberrant left subclavian artery is described. The patient had a right aortic arch. After a successful aortosubclavian artery bypass, symptoms due to brain ischemia disappeared. This is a very rare disease that is sometimes associated with an aortic anomaly, therefore the optimal therapeutic procedure need to be carefully selected, including the operative indications and approach. PMID- 10452253 TI - Stapled functional end-to-end anastomosis in Crohn's disease. AB - The aim of this study was to review our experience of stapled functional end-to end anastomosis following a bowel resection for Crohn's disease. Between 1988 and 1997, 62 patients underwent 63 stapled functional end-to-end anastomoses, including 54 ileocolonic, 8 ileoileal, and 1 colocolonic. One patient developed an anastomotic leak and required a laparotomy. There were no anastomotic complications in any other patients. The median duration of follow-up was 21 (rang: 4-122) months. Only one patient developed recurrence at the stapled ileocolonic anastomosis 35 months after the operation. However, this patient required no further surgical treatment. Stapled functional end-to-end anastomosis is associated with a low incidence of complications and a low incidence of early recurrence. This technique may therefore be the anastomotic configuration of choice following a bowel resection for Crohn's disease. PMID- 10452254 TI - The immunohistochemical expression of BCL-2 oncoprotein in colorectal adenocarcinoma. AB - The immunoreactivity for bcl-2 oncoprotein was determined in tumor tissue samples from 40 patients with colorectal adenocarcinoma. Positive immunoreactivity for bcl-2 oncoprotein was seen in 14 of the 40 tumor tissue samples (35%) and was found to be associated with a significantly higher incidence of synchronous liver metastasis (P = 0.043); however, there was no correlation between the immunoreactivity for bcl-2 oncoprotein and tumor size, depth of tumor invasion, lymph node metastasis, or clinical stage of the disease. Among 33 patients who had undergone curative resection, disease-free survival did not differ significantly between 10 with bcl-2-positive tumors and 23 with bcl-2-negative tumors (P = 0.498). The present study showed that the positive immunohistochemical expression of bcl-2 oncoprotein was associated with a significantly higher incidence of liver metastasis from colorectal cancer, but it had no effect on the disease-free survival of patients who had undergone curative resection. PMID- 10452255 TI - Expression of cholecystokinin-B/gastrin receptors in medullary thyroid cancer. AB - OBJECTIVE: To characterise the cholecystokinin (CCK) receptor subtypes in medullary thyroid cancer by measuring the expression of CCK-A and CCK-B/gastrin receptor mRNA. DESIGN: Open study. SETTING: Teaching hospital, Sweden. SUBJECTS: 6 patients with medullary thyroid cancer. INTERVENTION: Pentagastrin stimulation test and measurement of calcitonin concentration. Biopsy specimens were analysed using reverse transcription polymerase chain reaction (RT-PCR). MAIN OUTCOME MEASURE: Presence of CCK-A and CCK-B/gastrin receptors. RESULTS: All 6 patients with medullary thyroid cancer had positive pentagastrin tests preoperatively. CCK B/gastrin receptors but not CCK-A receptors were detected by RT-PCR in all six biopsy specimens. By contrast, no CCK receptors were found in normal thyroid tissues or in other thyroid tumours (follicular adenoma, papillary carcinoma, or anaplastic carcinoma). CONCLUSION: The presence of CCK-B/gastrin receptors in medullary thyroid tumours may have important clinical implications. PMID- 10452256 TI - Radical surgery for Budd-Chiari syndrome. Direct excision and repair for obstruction of the vena cava (Budd-Chiari syndrome) under hepatic vascular exclusion using a centrifugal pump. AB - OBJECTIVE: To attempt to reduce the incidence of hepatic and cardiac failure after radical surgery for the Budd-Chiari syndrome. DESIGN: Retrospective case study. SETTING: University hospital, Japan SUBJECTS: Three patients with obstruction of the vena cava (Budd-Chiari syndrome) by a thick membrane, thrombus, and a long stenosis, respectively. INTERVENTION: Direct excision and repair by patch dilatation under hepatic vascular exclusion using a Biomedicus centrifugal pump. RESULTS: Hepatic vascular exclusion provides good visibility and enables accurate assessment of the obstruction, proper treatment, and relatively little blood loss. There were no complications, particularly liver failure or heart failure, and the inferior vena cava stayed patent in all cases. Liver function was improved in all three patients. CONCLUSION: This technique is safe and reliable for removal of obstruction of the inferior vena cava (Budd Chiari syndrome). PMID- 10452257 TI - Major compared with limited hepatic resection for hepatocellular carcinoma without underlying cirrhosis: a retrospective analysis. AB - OBJECTIVE: To find out if patients with hepatocellular carcinoma (HCC) with no underlying cirrhosis benefit from major hepatic resection. DESIGN: Retrospective study. SETTING: University hospital, Japan. PATIENTS: 58 patients without cirrhosis and with HCC 10 cm in diameter or less. INTERVENTIONS: 25 had major and 33 had limited hepatic resections. MAIN OUTCOME MEASURES: Overall and disease free survival, and prognostic factors verified by univariate and multivariate analyses. RESULTS: 6 patients developed major complications (10%), two of whom died within 60 days of operation. There were no differences in postoperative morbidity and mortality between the two groups. The overall and disease-free survival were similar as was the incidence and pattern of intrahepatic tumour recurrence. Hepatitis B surface (HBs) antigen (positive), tumour size (smaller than 3 cm), and surgical margin (clear) were favourable indicators of disease free survival on multivariate analysis. CONCLUSIONS: Major hepatic resection should not necessarily be done for HCC without cirrhosis but it is important to take an adequate surgical margin. Overall and disease-free survival are better in patients who are HBs-antigen positive than those who are negative because most of the latter are positive for hepatitis C virus. PMID- 10452258 TI - Hepatic resection and percutaneous ethanol injection for the treatment of selected patients with more than one hepatocellular carcinoma. AB - OBJECTIVE: To report our results in a selected series of patients who had two separate hepatocellular carcinomas that were not suitable for treatment by resection alone or percutaneous injection of ethanol alone, whom we treated by a combination of the two. DESIGN: Open study. SETTING: Teaching hospital, Italy. SUBJECTS: 11 patients with Child's grade A or B cirrhosis who had two hepatocellular carcinomas in distant segments of the liver. INTERVENTIONS: Percutaneous injection of ethanol into the smaller of the two nodules before, during, or after resection. MAIN OUTCOME MEASURES: Morbidity, mortality, and recurrence. RESULTS: No patient died and there was one postoperative complication (atelectasis). During a median follow-up period of 24 months (range 8-48) two patients have developed recurrences (at 10 and 18 months). CONCLUSION: Although we have studied only a few patients for a relatively short time this double approach may be an important treatment option for suitable patients with two hepatocellular carcinomas. PMID- 10452259 TI - Intra-arterial 5-fluorouracil and intravenous folinic acid in the treatment of liver metastases from colorectal cancer. AB - OBJECTIVE: To compare two regimens of intra-arterial chemotherapy for the treatment of hepatic metastases from colorectal cancer. DESIGN: Open study. SETTING: Teaching hospital, UK SUBJECT: 57 patients with unresectable metastases confined to the liver, and an indwelling catheter in the hepatic artery. INTERVENTIONS: The first 33 patients had a 24-hour intra-arterial infusion of 5 fluorouracil (5-FU) 1500 mg/m2, together with folinic acid 200 mg/m2 intravenously for the first and last two hours of the 5-FU infusion. This was repeated at weekly intervals for six weeks followed by a two-week gap before the next cycle. The remaining 24 patients had a two-weekly regimen in which folinic acid 200 mg/m2 was infused intravenously over 2 hours followed by an intra arterial loading dose of 5-FU 400 mg/m2 over 15 minutes; 5-FU 1600 mg/m2 was then given by intra-arterial infusion over 22 hours. This was repeated on day 2 and then at two-weekly intervals. MAIN OUTCOME MEASURES: Response rate and toxicity. RESULTS: Median follow-up was 21 months, and estimated median survival 19 months. 29 patients (51%) have responded, 5 completely. There are no significant differences between the groups. Sites of progression were liver alone 26 (53%), lung alone 9 (18%), liver and lung 3 (6%), and the remainder in local or regional nodes (n = 7) or bone (n = 4). Six patients experienced WHO grade 3 or 4 toxicity. CONCLUSION: The two regimens have high response rates and cause little systemic toxicity. Intra-arterial chemotherapy for hepatic metastases from colorectal cancer is currently being compared with conventional systemic chemotherapy in a randomised controlled trial. PMID- 10452260 TI - Efficacy of monoclonal antibody 131I-B72.3 immunoscintigraphy of pancreatic adenocarcinoma xenografts in nude mice. AB - OBJECTIVE: To assess the efficacy of monoclonal antibody (MoAb) B72.3 for in vivo immunoscintigraphy of pancreatic carcinoma in nude mice. DESIGN: Experimental controlled animal study. SETTING: University hospital, The Netherlands. SUBJECTS: 11 nude mice with subcutaneously xenografted human pancreatic carcinoma. INTERVENTIONS: Specific MoAb B72.3 and non-specific MoAb MOPC21 were iodinated with 131I and injected intraperitoneally in nude mice. Scintigrams were taken on days 1-10 and tumour:non-tumour ratios of the regions of interest (tumour, thorax, abdomen, background) were calculated. The mice were then killed for in vitro tissue counts. MAIN OUTCOME MEASURES: Tumour:non-tumour ratios in vivo and in vitro. RESULTS: Results of immunoscintigraphy on days 1, 2, and 6 were compared. In the B72.3-group all ratios were only moderately raised, the tumour:background ratio being the highest (2.35 (SD 0.67)) on day 6. There were no obvious differences between the ratios of the B72.3-group and the MOPC21 group. The results of tissue counts done at the end of the study, showed that tumour:non-tumour ratios were twice as high in the B72.3-group, suggesting some specificity of this MoAb. CONCLUSION: The results of our study suggest that MoAb B72.3 is not powerful enough for in vivo detection of pancreatic cancer as assessed in this xenograft model in nude mice. PMID- 10452261 TI - Foreign body reaction to meshes used for the repair of abdominal wall hernias. AB - OBJECTIVE: To investigate the local tissue reactions to meshes that had been removed from humans. DESIGN: Open study SETTING: Surgical department of the technical University, Aachen, Germany. MATERIAL: Samples of 17 non-absorbable meshes (1 polyester, 10 polypropylene, 2 reduced polypropylene, and 4 polytetrafluorethylene, PTFE) and 1 absorbable mesh (polyglactin 910) that had been implanted for repair of abdominal wall defects. INTERVENTIONS: Light and transmission electron microscopy, immunohistochemistry, and histological examination. MAIN OUTCOME MEASURES: Signs of inflammatory response. RESULTS: Light microscopy showed chronic inflammatory tissue reaction, even after years, with pronounced differences among materials. Partial volume of inflammatory cells (%) varied from 32 in polypropylene, to 12 in expanded PTFE, 8 in polyester, and 7 in reduced polypropylene. Formation of connective tissue correlated significantly with the extent of the inflammatory reaction (p<0.01). In meshes implanted for long periods there were still numerous macrophages at the interface between tissue and polypropylene (45%), polyester (45%), expanded PTFE (25%), and reduced polypropylene (22%). There was no difference in time dependent tissue reactions (p = 0.19). CONCLUSION: Inflammation around alloplastic materials used to repair defects in the abdominal wall persists for many years. There was evidence of long term wound complications as a result of persistent foreign body reactions. Further studies are required to evaluate the long term tissue response to these materials. PMID- 10452262 TI - Complications and management of Meckel's diverticulum--a review. AB - OBJECTIVE: To review the anatomy, symptomatology, diagnosis, complications and management of symptomatic and asymptomatic Meckel's diverticula. DESIGN: Retrospective study. SETTING: Teaching hospital, The Netherlands. SUBJECTS: 27 consecutive patients who underwent resection of a Meckel's diverticulum during the 16 year period 1 January 1981-31 August 1997. MAIN OUTCOME MEASURES: The symptomatology and histopathological findings. RESULTS: 15 patients had symptoms and 12 did not. Haemorrhage and inflammation were the most common presenting symptoms. All histopathological signs of inflammation were found in those with symptomatic diverticula. Resected asymptomatic Meckel's diverticula were abnormal in only two occasions: one contained an adenocarcinoma and one a faecolith. CONCLUSION: All symptomatic Meckel's diverticula should be resected. The surgical treatment of asymptomatic Meckel's diverticula is still controversial. PMID- 10452263 TI - Diagnostic accuracy of inflammatory markers in patients operated on for suspected acute appendicitis: a receiver operating characteristic curve analysis. AB - OBJECTIVE: To analyse the diagnostic accuracy of inflammatory variables in patients operated on for suspected acute appendicitis. DESIGN: Open prospective population based study. SETTING: Teaching hospital, Norway. INTERVENTIONS: Appendicectomy in 544 patients with clinically suspected acute appendicitis. MAIN OUTCOME MEASURES: Diagnostic accuracy of inflammatory variables using receiver operating characteristic (ROC) curve analysis. Logistic regression model of inflammatory variables using results of ROC-analysis. RESULTS: A small area under the ROC curve (AUC) (between 0.56 and 0.69) indicated less diagnostic accuracy. The best cut-off values were associated with low sensitivity and specificity, varying from 46% to 88%. Age, duration of history, and histological grade of inflammation significantly influenced the test results (AUC >0.5). The white cell count (WCC) and C-reactive protein (CRP) concentration were independent predictors of acute appendicitis with cut-off values of >12.3x10(9)/L and >0 mg/L, respectively, but AUC values of over 0.5 were observed only in patients between 13 and 40 years of age. CONCLUSION: Inflammatory variables added information of limited value in the diagnosis of suspected acute appendicitis. The test results should be interpreted differently in different groups of patients. PMID- 10452264 TI - Transanal electrovaporisation of rectal tumours. AB - OBJECTIVE: To present our early results with endoscopic transanal electrovaporization of rectal tumours. DESIGN: Retrospective study. SETTING: Department of Surgery, Faculty of Medicine, University of Selcuk. SUBJECTS: 10 patients, 4 with malignant and 6 with benign lesions. INTERVENTIONS: Vaporization of the tumour using a new ridged roller electrode, the VaporTrode, and a standard 24F urological resectoscope. MAIN OUTCOME MEASURES: Morbidity and recurrence. RESULTS: There were no complications and all adenomas were vaporized completely. 2 patients with obstruction required a further treatment 3 and 5 months later because of recurrence. Median duration of operation was 22.5 minutes (range 10 38) and median duration of hospital stay was 3 days (range 1-4). CONCLUSION: The VaporTrode is a safe and potentially useful tool for transanal excision of rectal tumours, the long term efficacy of which should be tested in larger series of patients. PMID- 10452265 TI - The sialyl Lewis X analogue, CY-1503, down-regulates leucocyte-endothelium interactions and the inflammatory response. AB - OBJECTIVE: To elucidate mechanisms of protection of ischaemic liver with the sialyl Lewis X analogue CY-1503 by regulation of inflammatory mediators such as oxygen free radicals and cytokines as well as blocking the migration of leucocytes. DESIGN: Laboratory study. SETTING: Teaching hospital, Spain. ANIMALS: 122 male Sprague-Dawley rats divided into four groups: normal (n = 18), sham operated (n = 28), ischaemic controls (n = 38), and CY-1503 (n = 38). INTERVENTIONS: Warm total hepatic ischaemia for 90 minutes followed by various periods of reperfusion. MAIN OUTCOME MEASURES: Survival, liver histology, liver function, neutrophil infiltration, and free radical and cytokine concentrations. RESULTS: 2/20 ischaemic controls survived, compared with 14/20 given CY-1503. Liver function was better, as was histological appearance judged by the Suzuki score); myeloperoxidase activity was significantly decreased (n = 6 in each group, p<0.01) as were concentrations of free radicals (n = 12 in each group, p<0.05) in the group given CY-1503. CY-1503 had no effect on concentrations of the cytokines tumour necrosis factor-alpha or interleukin 1-alpha. CONCLUSIONS: CY-1503 exerts a protective effect in that it able to down-regulate concentrations of free radicals in our rat model. It is a potent inhibitor of neutrophil migration, but has no effect on cytokine concentrations. PMID- 10452266 TI - Postoperative exposure to glove powders modulates production of peritoneal eicosanoids during peritoneal wound healing. AB - OBJECTIVE: To assess the effect of postsurgical exposure of peritoneal cavity to glove powders, Hydrocote, latex proteins, and lipopolysaccharide (LPS) on eicosanoid production in peritoneal fluid and cellular distribution of eicosanoid enzymes in peritoneal wound during healing. DESIGN: Randomised experimental study. SETTING: Institute for Wound Research, USA. ANIMALS: 360 mice randomised into six groups of 60 each. INTERVENTION: Abrasion of peritoneal cavity followed by instillation of 500 microl of sterile phosphate buffered saline (PBS) alone (Control) or containing 100 microg/ml of Biosorb, Keoflo, Hydrocote, 1 mg/ml of latex proteins, or 12.5 microg/ml of LPS. Mice were killed at 1, 2, 4, 7, 14 and 28 days, and the peritoneal washing obtained from each animal and concentration of eicosanoids measured. Tissue were immunostained for cyclooxygenases and 5 lipoxygenase and thromboxane A2 (TXA2) synthetase. RESULTS: Peritoneal fluid from uninjured controls contained 3.9 (0.8), 5.2 (0.3) and 0.2 (0.02) ng/ml of thromboxane B2 (TXB2), prostaglandin E2 (PGE2) and leukotriene B4 (LTB4), respectively. These increased significantly during the first week to 6.3 (0.3), 11.7 (0.8) and 2.6 (0.1) ng/ml, p<0.05, before returning to baseline by day 14. In all the treated groups the values were significantly higher than in controls (p<0.05). Immunoreactive cyclo-oxygenases, 5-lipoxygenase and TXA2 synthetase proteins were present in various cell types in uninjured skin and peritoneum, incisional and peritoneal wounds and adhesion tissues. Staining was more intense at the site of wounds and paralleled eicosanoid concentrations during healing. There was no difference between exposed and unexposed groups. CONCLUSION: The presence of glove powders, latex proteins and LPS in peritoneal cavity cause increased eicosanoid production and aggravate the normal inflammatory reaction to tissue injury. This may contribute to the inflammatory or immune reactions and development of adhesions caused by glove powders. PMID- 10452267 TI - Intraperitoneal transplantation of isologous mesothelial cells for prevention of adhesions. AB - OBJECTIVE: To study the effect of transplantation of mesothelial cells on the formation of adhesions after peritoneal abrasion. DESIGN: Animal study. SETTING: Teaching hospital, Germany. ANIMALS: 30 isologous eight-week old Lewis rats to allow for harvesting of the greater omentum from a separate group. INTERVENTIONS: The first group (n = 10) served as mesothelial cell donors. The other animals had laparotomy and induction of adhesions by standardised abrasion of the peritoneum. The trial group (n = 10) were given a suspension of 10(6) mesothelial cells/100 g body weight intraperitoneally and the control group (n = 10) an equal volume of culture medium. After 10 days the animals were killed. MAIN OUTCOME MEASURES: Measurements of the areas of adhesions by computer aided morphometry. RESULTS: The trial group developed a mean (SD) adhesion area of 122.7 (176.7) mm2, and the controls 310.5 (179.1) mm2. The corresponding medians (range) were 51.2 (0-547.1) and 274.3 (100.6-575.4). Transplantation of mesothelial cells resulted in a significant reduction in adhesion formation (Wilcoxon test, p<0.01). CONCLUSION: Intraperitoneal transplantation of mesothelial cells is an effective way of reducing the formation of adhesions. PMID- 10452268 TI - Continuous single-layer anastomoses with monofilament poliglecaprone sutures in abdominal surgery. PMID- 10452269 TI - Spontaneous rupture of the spleen: role of ultrasound in diagnosis, treatment, and monitoring. PMID- 10452270 TI - Testicular vein tear by blunt trauma: anatomical considerations of a rare injury. PMID- 10452271 TI - Colorectal resection in the elderly by J.J. Arenal, C. Benito, M.P. Concejo and E. Ortega (Eur J Surg 1999; 165: 593-597): invited commentary. PMID- 10452272 TI - The burden of genetic disease and attitudes towards gene testing in Alport syndrome. AB - We evaluated the burden of Alport syndrome (AS) and the attitudes towards gene testing in patients with AS and their healthy family members from 37 families using a multiple-choice questionnaire. We also evaluated how the disease affected the decision to have children and the information received about the syndrome. A total of 53 individuals responded to this questionnaire. The risk of renal insufficiency and the uncertainty of the prognosis were considered the worst components of AS. Many of the respondents felt that children should be informed about AS as soon as they start asking (45%), preferably by a parent (74%). Almost all of the respondents (96%) had a positive attitude towards genetic research, which in the opinion of the majority should be aimed at better treatment and diagnosis of the disease rather than developing methods for prenatal diagnosis (89% and 75% versus 43%). AS seems to be well tolerated; 28% and 19% of the respondents found abortion acceptable in cases of an affected male and female fetus, respectively. Our study indicates a desire for prenatal tests in order to predict the health of a future child rather than for selective abortion. PMID- 10452273 TI - Hereditary glomerulopathy associated with a mitochondrial tRNA(Leu) gene mutation. AB - Several cases of hereditary glomerulopathy associated with an A to G transition at position 3243 in mitochondrial DNA, which is known to be associated with most cases of MELAS syndrome (myopathy, encephalopathy, lactic acidosis, and stroke like episodes), have been recently reported. These patients share the characteristics of hereditary progressive glomerular disease and hearing loss with Alport syndrome. We therefore screened 27 patients with kidney disease clinically mimicking Alport syndrome for the presence of the 3243 mitochondrial mutation, and found one girl with the mutation and a positive family history. Her clinical features were very similar to those of all cases reported to date. An absence of hematuria, severe kidney involvement in a female, pathological changes of focal segmental glomerulosclerosis with no basket-weave change of the glomerular capillary wall, and the frequent association of steroid-induced diabetes are the major features that distinguish this condition from Alport syndrome. Careful neurological examination may detect neuromuscular symptoms compatible with mitochondrial cytopathies. In conclusion, progressive glomerulopathy should be included in the broad spectrum of mitochondrial cytopathies, especially in cases of MELAS syndrome. This mutation should also be included in the etiologies of secondary focal segmental glomerulosclerosis and in the differential diagnosis of Alport syndrome. PMID- 10452274 TI - Autosomal dominant inheritance of multicystic dysplastic kidney. AB - Unilateral multicystic dysplastic kidney (MCDK) in a normal infant is believed to be a sporadic disorder, with an incidence of about 1 in 4,300 live births. Isolated unilateral MCDK occurring in a family without other genitourinary abnormalities has not been described. We report a family in which isolated unilateral MCDK occurred in a woman and her two children. The mother presented with a palpable abdominal mass during infancy, which on excision was found to be a MCDK. Both the children were found to have MCDK on prenatal ultrasonography, which was later confirmed on postnatal evaluation. The MCDK in the children continues to involute on follow-up urinary tract ultrasonography. The inheritance of MCDK appears to be autosomal dominant in this family. PMID- 10452275 TI - A patient with pseudohypoaldosteronism type 1 and respiratory distress syndrome. AB - We present a patient born at 36 weeks' gestation with respiratory distress, who required 6 days of mechanical ventilation, without a demonstrable infectious cause. He also developed hyponatremia, hypernatriuria, elevated serum aldosterone levels, and probable pseudohypoaldosteronism type 1 (PHA-1). This appears to be the first reported human case of both respiratory distress and a renal salt wasting process with elevated serum aldosterone. In animal models, abnormalities of subunits of the epithelial sodium channel produce respiratory distress and PHA 1. This patient's clinical presentation could be due to the same processes. PMID- 10452276 TI - Markers of endothelial cell activation and injury in childhood haemolytic uraemic syndrome. AB - Diarrhoea-associated haemolytic uraemic syndrome (D+ HUS) is usually caused by verotoxin-producing Escherichia coli. Histology shows endothelial swelling with localised thrombus. Activation of coagulation and fibrinolysis also occurs. These facts, combined with the knowledge that recovery usually follows within weeks, led us to hypothesise that verotoxin causes localised endothelial cell activation but not injury. Markers of endothelial cell activation and injury were measured serially in 30 children with acute D+ HUS, healthy children, and children receiving chronic dialysis. Interpretation of markers was complicated by the renal dysfunction characteristic of D+ HUS. Nevertheless there was no evidence for endothelial cell injury, as soluble tissue factor levels were not increased and soluble thrombomodulin levels were lower than dialysed controls (P<0.001). In the acute phase, soluble vascular cell adhesion molecule levels were raised above normal (P<0.001), but were lower than dialysed controls (P<0.001), and soluble E selectin levels were not significantly increased compared with normal controls (P=0.2). Hence, there was no evidence for endothelial cell damage or endothelial cell activation by the time children reached hospital; but this study did not exclude the possibility that endothelial cell activation occurred prior to hospital admission. PMID- 10452277 TI - ACE inhibition scintigraphy in the management of hypertension in children. AB - Angiotensin converting enzyme (ACE) inhibition scintirenography was performed to help establish the diagnosis and plan treatment of renovascular hypertension (RVH) in 57 hypertensive pediatric patients, 33 infants and 24 children older than 1 year. In 16 of 33 hypertensive infants, ACE inhibition scintirenography established the diagnosis of RVH from renal ischemia (due to aortic or renal arterial thrombi). Two scintigraphic criteria were used for the diagnosis of RVH: criterion I, ischemic and damaged kidney (a non-functioning kidney on or off ACE inhibition) and criterion II, ischemic but not damaged kidney (ACE inhibition induced deterioration of function of the kidney). When criterion I was present and the contralateral kidney was normal, ACE inhibitors could be used for treatment of hypertension without deterioration of renal function; kidneys satisfying criterion I eventually involuted or manifested growth arrest and frequently caused persistent RVH, even after resolution of the thrombus, requiring nephrectomy. When criterion II was present bilaterally, or it was associated with criterion I contralaterally, the use of antihypertensive drugs other than ACE inhibitors was necessary in order to prevent renal insufficiency or failure from ACE inhibitors. However, kidneys with criterion II showed normal growth and, following retraction or dissolution of the aortic thrombus, hypertension resolved. In 2 of 24 hypertensive children older than 1 year, the test was diagnostic of branch renal artery stenosis; RVH was cured by selective angioplasty. ACE inhibition scintirenography is useful in the evaluation and planning of treatment in children with hypertension and may predict the outcome of therapy and ultimate renal function. PMID- 10452278 TI - Diagnostic sensitivity of serum cystatin for impaired glomerular filtration rate. AB - Recently, the reciprocal of cystatin C (Cys-C), a non-glycosylated 13-kilodalton protein that is produced by all investigated nucleated cells, was found to correlate closely with glomerular filtration rate (GFR). In order to determine the diagnostic validity in children for the detection of impaired GFR, venous blood samples from 381 children (aged 1.7-18 years) with various renal pathology referred for 51Cr-EDTA clearance investigations were obtained for measurement of Cys-C as well as beta2-microglobulin (beta2-MG) and serum creatinine. Two hundred and sixteen children with clearance values >90 ml/min per 1.73 m2 constituted a control group, with a normal GFR. In the control group, Cys-C values were normally distributed with a mean of 0.94+/-0.27 mg/l and an upper reference limit (97.5th percentile) of 1.47 mg/l. In all children, there was a positive correlation between 51Cr-EDTA clearance and the reciprocal of Cys-C (r=0.64, P<0.0001), beta2-MG (r=0.59, P<0.0001), creatinine (r=0.55, P<0.0001), and the height/creatinine ratio (r=0.73, P<0.0001). Receiver-operating characteristics analysis showed that there were no significant differences between these three parameters for discriminating between patients with normal and reduced GFR, although there was a tendency towards the best diagnostic sensitivity of the GFR estimate according to the Schwartz formula. We conclude that for the detection of mildly impaired GFR, a full clearance study cannot be replaced by measurement of serum Cys-C or beta2-MG concentrations. PMID- 10452279 TI - Cystatin C as a marker for glomerular filtration rate in pediatric patients. AB - Cystatin C is a non-glycated 13-kilodalton basic protein produced by all nucleated cells. The low molecular mass and the basic nature of cystatin C, in combination with its stable production rate, suggest that the glomerular filtration rate (GFR) is the major determinant of cystatin C concentration in the peripheral circulation. Recently published studies have shown that cystatin C correlates more strongly than creatinine with GFR measured using the 51Cr-EDTA clearance. The aim of this study was to evaluate serum cystatin C as a marker for GFR in children. GFR was determined on medical indications using the 51Cr-EDTA technique in pediatric patients (2-16 years) in our renal unit. Simultaneously their cystatin C and creatinine concentrations were also measured. Of our 52 patients, 19 had a reduced renal function ( or =75% coronary artery narrowing was considered significant. In groups 2A and 2B, SPECT images were divided into 24 segments, and relative regional uptake in each segment was obtained. In group 2A, relative regional uptake of single 99mTc-sestamibi SPECT correlated well with that of dual isotope SPECT (r = 0.942). In group 2B, relative regional uptake of single 201TI SPECT correlated well with that of dual-isotope SPECT (r = 0.935). Furthermore, in low 201TI uptake segments with relative regional uptake in both single- and dual-isotope SPECT of < or =70%, the degree of concordance between single- and dual-rest 201TI was considered to be high with Bland-Altman analysis and the kappa statistic. Comparison of perfusion defect type demonstrated that, of 22 stress defects within infarct zones, 95% were irreversible and 5% were reversible. In contrast, of 28 stress defects within stenosed vessel zones in noninfarct zones, 89% were reversible and 11% were irreversible (P < 0.0001 versus infarct zones). CONCLUSION: Simultaneous dual-isotope imaging with Moore's correction method is feasible, with acceptable accuracy for detection of coronary artery disease and a small amount of crosstalk into each window. PMID- 10452304 TI - Kinetics of 123I-MIBG after acute myocardial infarction and reperfusion therapy. AB - Metaiodobenzylguanidine (MIBG) washout from the myocardium has been thought to reflect sympathetic nerve tone. After acute myocardial infarction, however, little is known about this parameter. The aim of this study was to determine the significance of cardiac washout after myocardial infarction and early reperfusion by investigating MIBG kinetics and correlating those kinetics to clinical parameters. METHODS: Sixty patients with acute myocardial infarction underwent planar MIBG and thallium imaging within 14 d of early reperfusion therapy. Global uptake and washout in myocardium, lungs and liver were calculated from early and delayed images. A regional analysis of myocardial kinetics in normal and infarcted myocardium and in an infarct border zone was also performed. Scintigraphic data were correlated with heart-rate variability as an electrophysiologic marker for autonomic tone and prevalence of arrhythmia in 52 patients. Heart-rate variability was described by time-domain indices from long term electrocardiogram recordings. An age-matched normal control group for MIBG consisted of 10 individuals without heart disease. RESULTS: The infarct patients had preserved left-ventricular ejection fraction (LVEF) (56% +/- 17%). Although late myocardial uptake was expectedly lower in infarct patients compared with healthy volunteers (2.36 +/- 0.66 versus 2.80 +/- 0.55; P = 0.04), global myocardial MIBG washout was faster (11.6% +/- 7.9% versus 0.2% +/- 10.2%, respectively; P = 0.002). Lung and liver kinetics did not differ in patients and healthy volunteers. Global MIBG washout showed a weak but significant positive correlation with the baseline heart rate (r = 0.28, P = 0.03) and an inverse correlation with LVEF (r = -0.28, P = 0.04). Washout was faster in a subgroup of 8 patients with reduced heart-rate variability (16.5% +/- 9.9% versus 10.3% +/- 8.3%; P = 0.04). Regional analysis revealed similar degrees of enhanced MIBG washout for infarcted (low perfusion, low MIBG uptake) and remote myocardium (normal perfusion, high MIBG uptake), whereas the border zone (normal perfusion, low MIBG uptake) showed a nonsignificant trend toward higher washout. CONCLUSION: After myocardial infarction, changes in MIBG kinetics occur specifically in the myocardium, whereas kinetics in lung and liver remain unchanged. Even in patients with left-ventricular function preserved by reperfusion therapy, MIBG washout is abnormal and globally increased. Enhanced washout may reflect increased sympathetic nerve tone and represent increased catecholamine turnover or impaired reuptake in the subacute phase of myocardial infarction. PMID- 10452305 TI - Sympathetic reinnervation of cardiac allografts evaluated by 123I-MIBG imaging. AB - Some heart-transplant patients present with improved heart rate response to exercise and anginal pain suggesting reinnervation of allografts. Studies performed up to 5 y post-transplantation have suggested that reinnervation is a slow process that occurs only after 1 y post-transplantation. The purpose of this study was to evaluate the extent of sympathetic reinnervation in heart-transplant patients and its relation to cardiac function. METHODS: We performed 123I metaiodobenzylguanidine (MIBG) studies and rest/exercise radionuclide ventriculography in 31 heart-transplant patients 6 mo to 12 y post transplantation. Intensity of myocardial MIBG uptake was quantified by a heart-to mediastinum ratio (HMR), and the regional distribution of MIBG was determined by tomographic studies. RESULTS: HMR correlated positively with time after transplantation (r = 0.607, P < 0.001). Patients studied from 2 to 12 y post transplantation had an HMR significantly higher than patients studied before 2 y post-transplantation (1.62 +/- 0.2 versus 1.34 +/- 0.2, P < 0.05). Myocardial MIBG uptake was anterolateral in 16 patients, anterior in 3 and anterolateral and septal in 3. Myocardial MIBG uptake was absent in 9 patients. Vasculopathy developed in 8 patients, and 5 of them (63%) had decreased myocardial MIBG uptake. Peak filling rate was higher in patients studied from 2 to 12 y post transplantation (2.7 +/- 0.8 end-diastolic volume (EDV)/s versus 2.16 +/- 0.5 EDV/s, P = 0.02). CONCLUSION: Sympathetic reinnervation increases with time after heart transplantation and is seen more frequently after 2 y post-transplantation. Complete reinnervation of the transplanted heart does not occur even up to 12 y post-transplantation. Early vasculopathy may delay the process of sympathetic reinnervation. PMID- 10452306 TI - Prognostic value of MIBG imaging in idiopathic dilated cardiomyopathy. AB - Alterations of cardiac sympathetic innervation are likely to contribute to fatal outcomes in patients with heart failure. These alterations can be evaluated noninvasively by 123I-metaiodoben-zylguanidine (MIBG) imaging. METHODS: The hypothesis that impaired cardiac sympathetic innervation, as assessed using MIBG imaging, is related to adverse outcomes was tested in 112 patients with heart failure resulting from idiopathic cardiomyopathy. Main inclusion criteria were New York Heart Association classes II-IV and radionuclide left ventricular ejection fraction (LVEF) < 40%. Patients were assessed for cardiac MIBG uptake, circulating norepinephrine concentration, LVEF, peak Vo2, x-ray cardiothoracic ratio, M-mode echographic end-diastolic diameter and right-sided heart catheterization parameters. RESULTS: During a mean follow-up of 27 +/- 20 mo, 19 patients had transplants, 25 died of cardiac death (8 sudden deaths), 2 died of noncardiac death and 66 survived without transplantation. The only independent predictors for mortality were low MIBG uptake (P < 0.001) and LVEF (P = 0.02) when using multivariate discriminant analysis. Moreover, MIBG uptake (P < 0.001) and circulating norepinephrine concentration (P = 0.001) were the only independent predictors for life duration when using multivariate life table analysis. CONCLUSION: Impaired cardiac adrenergic innervation as assessed by MIBG imaging is strongly related to mortality. MIBG imaging may help risk stratify patients with heart failure resulting from idiopathic dilated cardiomyopathy. PMID- 10452307 TI - Influence of arrhythmias on gated SPECT myocardial perfusion and function quantification. AB - Despite the importance of R-wave gating myocardial perfusion tomography for ventricular function assessment, neither prevalence of gating errors nor their influence on quantified cardiac parameters has been studied. METHODS: Arrhythmia induced anomalies in curves of counts versus projection angle for each R-wave segment were detected visually and algorithmically. Arrhythmia prevalence was tabulated for 379 patients (group 1) with prospective coronary artery disease (mean age 63+/-13 y, 47% male). Myocardial counts were analyzed from all reconstructed cinematic midventricular slices to assess arrhythmia effects on percentage of systolic count increase, generally assumed to equal percentage of wall thickening. In a separate retrospective analysis of 41 patients (group 2), with coronary artery disease (mean age 64+/-12 y, 68% male) having no significant arrhythmias, 36 of whom also underwent equilibrium radionuclide angiography, original projection data were altered to simulate arrhythmia-induced aberrant count patterns to evaluate effects on ventricular function and perfusion measurements. RESULTS: Group 1 patients consisted of 26% without gating errors, 32% with count losses only in the last R-wave interval due to inconsistent transient increase of heart rate, 24% with count decreases in several late intervals due to consistently variable rates, 8% with early interval count increases paired with late interval count decreases due to ectopic beats and 9% with erratic count changes due to atrial fibrillation. Observed count patterns were strongly associated (P < 10(-3)) with arrhythmias detected by electrocardiogram monitoring. In group 2 simulations, ventricular volumes changed by only 2%+/-9% and ejection fraction (EF) by only 1%+/-4% from control values and correlated linearly (r> or = 0.96) with control values for all simulated arrhythmias. SPECT and equilibrium radionuclide angiography EFs correlated similarly (r = 0.85-0.89) for control and all simulations. Percentage changes from control in perfusion defect extent and severity were larger than processing reproducibility limits, the largest change being for atrial fibrillation. Control wall thickening was 38%+/-17%, significantly lower (P < 10(-6)) than for simulated arrhythmias, reflecting similar observations for group 1 patients. CONCLUSION: Even though ventricular volumes and EFs were affected minimally by arrhythmias, both perfusion analysis and wall thickening were compromised. Consequently, quality assurance of gating may be critically important for obtaining accurate quantified parameters. PMID- 10452308 TI - In vivo imaging of muscarinic cholinergic receptors in temporal lobe epilepsy with a new PET tracer: [76Br]4-bromodexetimide. AB - Muscarinic acetyl cholinergic receptors (mAChRs) may be involved in the pathophysiology of partial epilepsy. Previous experimental and imaging studies have reported medial temporal abnormalities of mAChR in patients with medial temporal lobe epilepsy (MTLE). Suitable radiotracers for mAChR are required to evaluate these disturbances in vivo using PET. Dexetimide is a specific mAChR antagonist that has been labeled recently with 76Br. This first study in humans focused on regional distribution and binding kinetics of [76Br]4-bromodexetimide (BDEX) in patients with MTLE. METHODS: Ten patients with well-lateralized MTLE had combined MRI, 18F-fluorodeoxyglucose (FDG) PET and 76Br-BDEX PET studies. Time-activity curves were generated in PET-defined regions of interest, including the medial, polar and lateral regions of the temporal lobe; the basal ganglia; the external and medial occipital cortex; and the white matter. RESULTS: The highest radioactivity concentration was observed in the basal ganglia and in the cortical regions, whereas radioactivity was lower in the white matter. On late images of PET studies, 76Br-BDEX uptake was statistically significantly decreased only in the medial temporal region ipsilateral to the seizure focus (1.37 +/ 0.28, P < 0.01) as determined by FDG PET imaging, anatomic MRI and electroencephalogram correlation, compared with the contralateral medial temporal region (1.46 +/- 0.31). CONCLUSION: 76Br-BDEX concentration is reduced in the temporal lobe ipsilateral to the seizure focus in patients with MTLE. This preliminary study suggests that 76Br-BDEX is a suitable radiotracer for studies of mAChR in humans. Further studies are required to investigate the potential value of 76Br-BDEX PET in other neurological disorders with muscarinic disturbances. PMID- 10452310 TI - Technetium HMPAO SPECT study in dementia with Lewy bodies, Alzheimer's disease and idiopathic Parkinson's disease. AB - The aim of this study was to compare the regional cerebral blood flow measurements studied by SPECT in dementia with Lewy bodies (DLB) and Alzheimer's disease (AD) to determine the contribution of SPECT to the differential diagnosis of these two diseases. METHODS: SPECT analysis with 99mTc-hexamethyl propyleneamine oxime (HMPAO) was performed in 20 patients with probable DLB, 20 patients with probable AD and 20 patients with idiopathic Parkinson's disease (IPD). Ten pairs of regions of interest were analyzed. Tracer uptake was expressed as a corticocerebellar activity ratio. RESULTS: Compared with IPD, in the DLB group there was a global decrease of HMPAO uptake in cortical regions of interest except in the posterior frontal and occipital regions; in the AD group there was limited left temporal and parietal hypoperfusion. In the DLB group, frontal HMPAO uptake was significantly lower than in the AD group. Two predictive scores were established by a factorial discriminant analysis from six left cortical indices (medial frontal, lateral frontal, posterior frontal, temporoparietal, parietal and parietooccipital) and the Mini-Mental State Examination, which correctly classified 53 of 60 patients (88%) (DLB, 18 of 20; AD, 16 of 20; IPD, 19 of 20). CONCLUSION: These findings indicate the presence of diffuse cortical abnormalities in DLB and suggest that SPECT may be useful in discriminating in vivo DLB from AD, revealing mainly frontal hypoperfusion in the former group. We estimate that SPECT study increases the possibility of separating DLB and AD because both disorders share different patterns of cerebral blood flow abnormality. PMID- 10452309 TI - Global spatial normalization of human brain using convex hulls. AB - Global spatial normalization transforms a brain image so that its principal global spatial features (position, orientation and dimensions) match those of a standard or atlas brain, supporting consistent analysis and referencing of brain locations. The convex hull (CH), derived from the brain's surface, was selected as the basis for automating and standardizing global spatial normalization. The accuracy and precision of CH global spatial normalization of PET and MR brain images were evaluated in normal human subjects. METHODS: Software was developed to extract CHs of brain surfaces from tomographic brain images. Pelizzari's hat to-head least-square-error surface-fitting method was modified to fit individual CHs (hats) to a template CH (head) and calculate a nine-parameter coordinate transformation to perform spatial normalization. A template CH was refined using MR images from 12 subjects to optimize global spatial feature conformance to the 1988 Talairach Atlas brain. The template was tested in 12 additional subjects. Three major performance characteristics were evaluated: (a) quality of spatial normalization with anatomical MR images, (b) optimal threshold for PET and (c) quality of spatial normalization for functional PET images. RESULTS: As a surface model of the human brain, the CH was shown to be highly consistent across subjects and imaging modalities. In MR images (n = 24), mean errors for anterior and posterior commissures generally were <1 mm, with SDs < 1.5 mm. Mean brain dimension errors generally were <1.3 mm, and bounding limits were within 1-2 mm of the Talairach Atlas values. The optimal threshold for defining brain boundaries in both 18F-fluorodeoxyglucose (n = 8) and 15O-water (n = 12) PET images was 40% of the brain maximum value. The accuracy of global spatial normalization of PET images was shown to be similar to that of MR images. CONCLUSION: The global features of CH-spatially normalized brain images (position, orientation and size) were consistently transformed to match the Talairach Atlas in both MR and PET images. The CH method supports intermodality and intersubject global spatial normalization of tomographic brain images. PMID- 10452311 TI - Relationship among 201T1 uptake, nuclear DNA content and clinical behavior in metastatic thyroid carcinoma. AB - A prospective study of 201TI uptake was performed to compare 201TI uptake with nuclear deoxyribonucleic acid (DNA) content and clinical behavior of tumors in metastatic thyroid carcinoma and to assess the significance of 201TI uptake in evaluating clinical characteristics of thyroid carcinoma. METHODS: Fifty-six patients with metastases of differentiated thyroid carcinoma had 201TI scintigraphy. Grade of 201TI uptake was semiquantitatively assessed according to tumor-to-background ratio on 2-h late scan. Nuclear DNA content was analyzed within 3 wk of 201TI study by flow cytometry using biopsy material and was classified as diploidy or aneuploidy. Patients were followed up to examine incidence of tumor growth and/or anaplastic transformation. RESULTS: DNA content was diploidy in 48 patients and aneuploidy in 8 patients. 201TI uptake in the DNA aneuploid group (2.61 +/- 0.29) was significantly higher than that in the DNA diploid group (1.82 +/- 0.35, P < 0.01 for both groups). Tumor growth was observed in all patients with DNA aneuploidy but in only 5 of 48 patients with DNA diploidy (P < 0.01). Anaplastic transformation was observed in 3 patients in the DNA-aneuploid group but in none of the patients in the DNA-diploid group. CONCLUSION: High 201TI uptake indicates greater incidence of abnormal DNA content with aggressive clinical behavior of metastatic tumors. Thus, 201TI scintigraphy may be useful in characterizing metastatic thyroid carcinoma and in identifying those patients with poorer prognoses. PMID- 10452313 TI - Relative 99mTc-MAG3 renal uptake: reproducibility and accuracy. AB - The aim of this study was to estimate the reproducibility and accuracy of 99mTc mercaptoacetyltriglycine (MAG3) relative percentage uptake. METHODS: Reproducibility was evaluated on healthy volunteers who were submitted twice to a 99mTc-MAG3 renographic study, which used different uptake algorithms, different background corrections and different time intervals. Accuracy was evaluated in a group of patients with symmetrical or asymmetrical relative renal function, who underwent both 99mTc-dimercaptosuccinic acid (DMSA) and 99mTc-MAG3 studies, using the DMSA relative percentage uptake as a reference. RESULTS AND CONCLUSION: The methods that combined the best reproducibility and accuracy for estimating 99mTc MAG3 left-to-right uptake ratio were the integral method, with subrenal or perirenal background correction, and the Patlak-Rutland plot. The use of the integral method without background correction introduced a systematic bias, whereas the slope method resulted in high variability. Therefore these methods cannot be recommended. PMID- 10452312 TI - Renal function after tumor enucleation: assessment by quantitative SPECT of 99mTc dimercaptosuccinic acid uptake by the kidneys. AB - The purpose of this study was to evaluate the amount of functioning renal mass removed and the amount of remaining individual renal function after tumor enucleation. METHODS: Renal functional volume, percentage injected dose (%ID) per cubic centimeter of renal tissue and individual renal uptake of 24 operated and 24 contralateral kidneys were studied by two sequential SPECT quantitations of 99mTc-dimercaptosuccinic acid (DMSA) uptake by the kidneys (QDMSA). The first study was before surgery and the second study was 1 to 6 mo (mean 3.5 mo) after surgery. Mean tumor size was 3.4 +/- 0.99 cm and all tumors were confined to the renal parenchyma (stages pT1 and pT2). RESULTS: In the operated kidneys, there was a statistically significant decrease in renal cortical volume (170 +/- 46 mL after surgery versus 207 +/- 45 mL before surgery, t = 6.2, P < 0.001) and individual renal uptake (10.3% +/- 3.0% after surgery versus 13.0% +/- 2.9% before surgery, t = 5.4, P < 0.001). There was no statistically significant change after surgery compared with before surgery in the %ID per cubic centimeter of renal tissue of the operated kidneys, and in the volume, %ID per cubic centimeter and uptake of the contralateral normal kidneys. CONCLUSION: The results suggest that QDMSA is a noninvasive method able to assess changes in separate renal function. The limited functioning parenchymal loss after tumor enucleation had no effect on the opposite kidneys. PMID- 10452314 TI - Quantification of glucose transport and phosphorylation in human skeletal muscle using FDG PET. AB - PET with 2-[18F]-fluoro-2-deoxy-D-glucose (FDG) is used for quantifying glucose metabolism in brain and myocardium in vivo. We developed and validated a similar procedure for the quantification of the two initial steps of glucose metabolism in skeletal muscle in vivo. METHODS: The measurement protocol was first optimized by computer simulations. In addition to the accuracy in sampling plasma input and tissue time-activity curves, precise determination of the fractional blood volume, that is, the extracellular tissue volume fraction, plays a key role in correctness of the determined model constants. The optimized protocol was subsequently used to estimate transmembrane muscular glucose transport and hexokinase activity in six human subjects with normal or altered glucose utilization. PET was performed during the steady state of an euglycemic hyperinsulinemic clamp. RESULTS: A three-compartment model provides a better description of the experimental data than a two- or four-compartment model. Glucose clearance from the extracellular compartment into the skeletal muscle cell (K1) ranges from 0.024 to 0.093 mL/g/min. The intracellular glucose phosphorylation rate (k3) varies between 0.030 and 0.142 min(-1). The regional muscular glucose utilization, as calculated from the determined model parameters, lies between 10.7 and 83.3 micromol/kg/min and correlates with the whole-body glucose utilization as independently determined (R2 = 0.83; P < or = 0.01). CONCLUSION: We demonstrate by computer simulations that a three-compartment model can be used to characterize the first two steps of glucose metabolism in skeletal muscle. An optimized measurement protocol is developed and applied to experimental data. This experimental approach should be appropriate to test whether glucose transport or hexokinase activity is altered in disorders of muscular glucose utilization. PMID- 10452316 TI - Role of FDG PET in metastatic thyroid cancer. PMID- 10452315 TI - Value of FDG PET in papillary thyroid carcinoma with negative 131I whole-body scan. AB - The management of metastatic thyroid carcinoma patients with a negative 131I scan presents considerable problems. Fifty-four athyrotic papillary thyroid carcinoma patients whose 1311 whole-body scans were negative underwent 18F fluorodeoxyglucose (FDG) PET; the purpose was to determine whether this procedure could localize metastatic sites. We also assessed its usefulness in the management of these patients. METHODS: Whole-body emission scan was performed 60 min after the injection of 370-555 MBq 18F-FDG, and additional regional attenuation-corrected scans were obtained. Metastasis was pathologically confirmed in 12 patients and was confirmed in other patients by overall clinical evaluation of the findings of other imaging studies and of the subsequent clinical course. RESULTS: In 33 patients, tumor had metastasized, whereas 21 patients were in remission. FDG PET revealed metastases in 31 patients (sensitivity 93.9%), whereas thyroglobulin levels were elevated in 18 patients (sensitivity 54.5%). FDG PET was positive in 14 of 15 metastatic cancer patients with normal thyroglobulin levels. In 20 of 21 patients in remission, FDG PET was negative (specificity 95.2%), whereas thyroglobulin levels were normal in 16 patients (specificity 76.1%). The sensitivity and specificity of FDG PET were significantly higher than those of serum thyroglobulin. In patients with negative 1311 scans, FDG PET detected cervical lymph node metastasis in 87.9%, lung metastasis in 27.3%, mediastinal metastasis in 33.3% and bone metastasis in 9.1%. In contrast, among 117 patients with 131I scan-positive functional metastases, 131I scan detected cervical lymph node metastasis in 61.5%, lung metastasis in 56.4%, mediastinal metastasis in 22.2% and bone metastasis in 16.2%. In all 5 patients in whom thyroglobulin was false-negative with negative antithyroglobulin antibody, PET showed increased 18F-FDG uptake in cervical lymph nodes, mediastinal lymph nodes, or both. Among patients with increased 18F-FDG uptake only in the cervical lymph nodes, the nodes were dissected in 11. Metastasis was confirmed in all, even in normal-sized lymph nodes. CONCLUSION: FDG PET scan localized metastatic sites in 131I scan-negative thyroid carcinoma patients with high accuracy. In particular, it was superior to 131I whole-body scan and serum thyroglobulin measurement for detecting metastases to cervical lymph nodes. FDG PET was helpful for determining the surgical management of these patients. PMID- 10452317 TI - Potential applications of PET imaging in developing novel cancer therapies. PMID- 10452318 TI - Comparison of dual-head coincidence gamma camera FDG imaging with FDG PET in detection of breast cancer and axillary lymph node metastasis. AB - Dual-head coincidence gamma camera 18F-fluorodeoxyglucose (FDG) imaging was compared with FDG PET in the detection of breast cancer and axillary lymph node metastasis. METHODS: Both coincidence gamma camera FDG imaging and FDG PET were performed in a cylindrical phantom containing spheres of different sizes and activity ratios (5:1, 10:1 and 15:1) and in 30 women (age range 32-78 y) with suspected breast cancer. Biopsies or mastectomies were performed in all patients. Images were visually assessed, and the count ratio between tumor and normal tissue (T/N ratio) was calculated. RESULTS: In the phantom studies, coincidence gamma camera imaging visualized the smallest sphere (1.0 cm) at a ratio of 15:1 but not at ratios of 5:1 and 10:1. Coincidence gamma camera imaging visualized the other spheres (> or =1.3 cm) at all ratios. PET visualized all spheres at all ratios. In the clinical studies, 22 of 26 breast carcinomas detected by PET were also detected by coincidence gamma camera imaging.. Coincidence gamma camera imaging detected all of the carcinomas > or =2 cm in diameter (n = 10) and 12 of 16 carcinomas <2 cm. In breast carcinomas detected by both PET and coincidence gamma camera imaging, the T/N ratio in non-attenuation-corrected PET (7.12 +/- 7.13) was significantly higher than in coincidence gamma camera imaging (2.90 +/- 1.47, P < 0.005). Four of 8 axillary lymph node metastases detected by PET were detected by coincidence gamma camera imaging. Of 9 axillary lymph node metastases <1.0 cm in diameter, 7 and 3 were detected by PET and coincidence gamma camera imaging, respectively. CONCLUSION: Coincidence gamma camera imaging is useful in detecting breast carcinoma > or =2 cm in diameter but is not reliable for breast carcinoma <2 cm in diameter. Coincidence gamma camera imaging may be useless or even dangerous in the detection of axillary lymph node metastasis. PMID- 10452319 TI - A new look at breast cancer. PMID- 10452320 TI - Effects of scatter substraction on detection and quantitation in hepatic SPECT. AB - The purpose of this investigation was to examine the effects of subtractive scatter compensation methods on lesion detection and quantitation. METHODS: Receiver operating characteristic (ROC) methodology was used to measure human observer detection accuracy for tumors in the liver using synthetic images. Furthermore, ROC results were compared with mathematical models for detection and activity quantitation to examine (a) the potential for predicting human performance and (b) the relationship between the detection and quantitation tasks. Images with both low and high amounts of scatter were compared with the ideal case of images of primary photons only (i.e., perfect scatter rejection) and with images corrected by subtracting a scatter image estimated by the dual photopeak window method. RESULTS: With low contrast tumors in a low count background, the results showed that scatter subtraction improved quantitation but did not produce statistically significant increases in detection accuracy. However, primary images did produce some statistically significant improvements in detection accuracy when compared with uncorrected images, particularly for high levels of scatter. CONCLUSION: Although scatter subtraction methods may provide improved activity quantitation, they may not significantly improve detection for liver SPECT. The results imply that significant improvement in detection accuracy for the conditions tested may depend on the development of gamma cameras with better scatter rejection. PMID- 10452321 TI - Bremsstrahlung radiation exposure from pure beta-ray emitters. AB - With increasing therapeutic use of radionuclides that emit relatively high-energy (>1 MeV) beta-rays and the production in vivo of bremsstrahlung sufficient for external imaging, the potential external radiation hazard warrants evaluation. METHODS: The exposure from a patient administered beta-ray-emitting radionuclides has been calculated by extending the National Council on Radiation Protection and Measurement model of a point source in air to account for biologic elimination of activity, the probability of bremsstrahlung production in vivo and its mean energy and the absorption by the patient's body of the bremsstrahlung thus produced. To facilitate such calculations, a quantity called the "specific bremsstrahlung constant" (in C/kg-cm2/MBq-h), betaBr, was devised and calculated for several radionuclides. The specific bremsstrahlung constant is the bremsstrahlung exposure rate (in C/kg/h) in air at 1 cm from a 1 MBq beta-ray emitter of a specified maximum beta-ray energy and frequency of emission in a medium of a specified effective atomic number. RESULTS: For pure beta-ray emitters, the retained activities at which patients can be released from medical confinement (i.e., below which the effective dose equivalent at 1 m will not exceed the maximum recommended value of 0.5 cSv for infrequently exposed members of the general public) are extremely large: on the order of hundreds of thousands to millions of megabecquerels. CONCLUSION: Radionuclide therapy with pure beta ray emitters, even high-energy beta-ray emitters emitted in bone, does not require medical confinement of patients for radiation protection. PMID- 10452322 TI - Radiolabeled peptides for targeting cholecystokinin-B/gastrin receptor-expressing tumors. AB - The high sensitivity of pentagastrin stimulation in detecting primary or metastatic medullary thyroid cancer (MTC) suggests widespread expression of the corresponding receptor type on human MTC. Indeed, autoradiographic studies have demonstrated cholecystokinin (CCK)-B/gastrin receptors not only in more than 90% of MTCs but also in a high percentage of small cell lung cancers, some ovarian cancers, astrocytomas and potentially a variety of adenocarcinomas. The aim of this study was to systematically screen and optimize, in a preclinical model and a pilot clinical study, suitable radioligands for targeting CCK-B receptors in vivo. METHODS: A variety of CCK/gastrin-related peptides, all bearing the C terminal CCK receptor-binding tetrapeptide sequence Trp-Met-Asp-PheNH2 or derivatives thereof, were studied. They were radioiodinated by the lodogen or Bolton-Hunter procedures. The peptides were members of the gastrin or CCK families, which differ by the intramolecular position of a tyrosyl moiety. Their stability and affinity were studied in vitro and in vivo; their biodistribution and therapeutic efficacy were tested in nude mice bearing subcutaneous human MTC xenografts. Diethylenetriamine pentaacetic acid (DTPA) derivatives of suitable peptides were synthesized successfully, and their preclinical and initial clinical evaluations were performed, labeled with 111In. RESULTS: All members of the CCK or gastrin families were stable in serum (with half-lives of several hours at 37 degrees C); nevertheless, the stability of those peptides bearing N terminal pGlu residues or D-amino acids was significantly higher. In accordance with their comparably low affinity, nonsulfated members of the CCK family showed fairly low uptake in the tumor and other CCK-B receptor-expressing tissues. Sulfated CCK derivatives performed significantly better but also displayed a comparably high uptake in normal CCK-A receptor-expressing tissues. This effect was probably due to their similar affinity for both CCK-A and CCK-B receptors. Best tumor uptake and tumor-to-nontumor ratios were obtained with members of the gastrin family because of their selectivity and affinity for the CCK-B receptor subtype. Pilot therapy experiments in MTC-bearing animals showed significant antitumor efficacy compared with untreated controls. DTPA derivatives of minigastrin were successfully developed. In a pilot clinical study, radioiodinated and 111In-labeled derivatives showed excellent targeting of physiological CCK-B receptor-expressing organs, as well as all known tumor sites. CONCLUSION: CCK/gastrin analogs may be a useful new class of receptor-binding peptides for diagnosis and therapy of CCK-B receptor-expressing tumors, such as MTC or small cell lung cancer. Nonsulfated gastrin derivatives may be preferable because of their CCK-B receptor selectivity, hence lower accretion in normal CCK A receptor-expressing organs. PMID- 10452323 TI - Quantification of myocardial blood flow using 13N-ammonia and PET: comparison of tracer models. AB - Several tracer kinetic methods have been proposed for quantification of regional myocardial blood flow (MBF) with 13N-ammonia and PET. Merits and limitations specific to each approach, however, generally are not clear, because they have not been evaluated in the same experimental environment. Therefore, we compared six different commonly used methods (11 modifications) to characterize the accuracy of each approach. The methods included the two-parameter model (method 1), the modified two-parameter model (method 2), the four-parameter model (method 3), the graphical analysis (method 4), the first-pass extraction method (method 5) and the dose uptake index (DUI; method 6). METHODS: Eleven studies in four dogs, 16 studies in eight healthy human volunteers and 14 studies in seven patients were performed using 13N-ammonia and PET. MBF in dogs was varied with dipyridamole and coronary occlusions and was measured independently and simultaneously with microspheres. Volunteers and patients were studied at baseline and after dipyridamole. MBF and DUI were estimated using a time-activity curve (Qi[t]) derived from dynamic images and regions of interest (ROls) and using the six methods. DUI was defined as Qi(t = 2 min) x weight/dose. RESULTS: MBF estimated by methods 1-5 correlated well with microsphere MBF in dogs. MBF estimates by method 1 correlated well with those by methods 2, 4 and 5 and to a lesser degree with those by method 3 in both dog and human studies. DUI correlated poorly with MBF by microspheres and by methods 1-5 in both dog and human studies. MBF estimates by method 3 showed larger dispersion (SD/mean flow) and higher sensitivity to metabolites correction in arterial blood than those by methods 1, 2, 4 and 5. CONCLUSION: MBF can be measured accurately using 13N ammonia PET and tracer kinetic methods. DUI is a poor indicator of MBF values. The results indicate that preference should be given to the two-parameter model, incorporating geometrical ROI representation (method 2) among the compartment models, and to the graphical analysis (method 4) among the noncompartmental approaches. PMID- 10452324 TI - Evaluation of glutathione localization in brain using 99mTc meso-HMPAO. AB - The relationship between distribution of 99mTc meso-hexamethyl propyleneamine oxime (HMPAO) and glutathione (GSH) content was studied in the mouse. METHODS: The regional distributions of 99mTc meso-HMPAO and 99mTc d,I-HMPAO were examined using tissue sampling and autoradiographic methods and were compared with GSH content and distribution by a histochemical procedure. RESULTS: The uptake of 99mTc meso-HMPAO was highest in the cerebellum and lowest in the brain stem, whereas the distribution of 99mTc d,I-HMPAO was more uniform. The regional distribution of 99mTc meso-HMPAO in the mouse brain correlated with GSH content (r = 0.787), but that of 99mTc d,I-HMPAO did not. Treatment with diethyl maleate, a GSH depletor, significantly decreased the 99mTc meso-HMPAO uptake to 21%-33% of the control in every region, but the reduction of the 99mTc d,I-HMPAO uptake was moderate (58%-65% of the control). In the autoradiograph, the radioactivity of 99mTc meso-HMPAO was higher in the gray matter than in the white matter of cerebellum, and more radioactivity was found in cerebellum and in hippocampus than in forebrain without the hippocampus. This pattern of distribution was similar to the histochemical localization of GSH estimated with a sulfhydryl reagent, Mercury orange. CONCLUSION: 99mTc meso-HMPAO might be used as an imaging agent to assess GSH localization in the brain. PMID- 10452325 TI - Radiolabeled alpha(v)beta3 integrin antagonists: a new class of tracers for tumor targeting. AB - The alpha(v)beta3 integrins play an important role during tumor metastasis and tumor-induced angiogenesis. Targeting of this receptor may provide information about the receptor status of the tumor and enable specific therapeutic planning. Cyclo(-Arg-Gly-Asp-D-Phe-Val-) has been shown to be a selective alpha(v)beta3 integrin antagonist with high affinity. In this study we describe the synthesis and biological evaluation of [125I]-3-iodo-D-Tyr4-cyclo(-Arg-Gly-Asp-D-Tyr-Val-) ([125I]P2), [125I]-3-iodo-Tyr5-cyclo(-Arg-Gly-Asp-D-Phe-Tyr-) ([125I]P4) and the negative control peptide [1251]-3-iodo-D-Tyr4-cyclo(-Arg-D-Ala-Asp-Tyr-Val-) ([125I]P6). METHODS: Peptides were assembled on a solid support using fluorenylmethoxycarbonyl amino acid coupling protocols. Radioiodination was performed using the iodogen method. The in vitro binding assays were performed using isolated, immobilized alphaIIbeta3 and alpha(v)beta3 integrins. Expression of the alphaVbeta3 receptor on the different tumors was validated by immunohistochemical methods using alpha(v) and alpha(v)beta3 specific antibodies. For biodistribution studies, nude mice with melanoma M21 or mammary carcinoma MaCaF and BALB/c mice with osteosarcoma were used. RESULTS: The in vitro binding assays demonstrate that the introduction of tyrosine and subsequent iodination have no influence on the high affinity and selectivity for alpha(v)beta3. Immunohistochemical staining clearly indicates the presence of the alpha(v)beta3 integrins on the tumor tissue of the melanoma and the osteosarcoma. Pretreatment and displacement studies show specific binding of [125I]P2 on melanoma M21 bearing nude mice and osteosarcoma-bearing BALB/c mice but less specific binding on mammary carcinomas. [125I]P2 exhibits fast elimination kinetics. The accumulation in the tumor 10 min postinjection is 2.07 +/- 0.32 %ID/g for the melanoma M21 and 3.50 +/- 0.49 %ID/g for the osteosarcoma and decreases to 1.30 +/- 0.13 %ID/g and 2.03 +/- 0.49 %ID/g 60 min postinjection, respectively. [125I]P4 shows even faster elimination kinetics, resulting in a tumor accumulation of 0.40 +/- 0.10 %ID/g 60 min postinjection for the osteosarcoma bearing BALB/c mice. Both peptides reveal predominately hepatobiliary excretion. For [1251]P2, this also is confirmed by autoradiography. The negative control peptide [125I]P6 shows no specific activity accumulation. CONCLUSION: [125I]P2 exhibits high affinity and selectivity for the alpha(v)beta3 integrin in vitro and in vivo and, thus, represents the first radiolabeled alpha(v)beta3 antagonist for the investigation of angiogenesis and metastasis in vivo. PMID- 10452326 TI - Characterization of [18F]fluoroetanidazole, a new radiopharmaceutical for detecting tumor hypoxia. AB - Fluorinated derivatives of etanidazole are being explored as probes for tumor hypoxia. Our research group has synthesized [18F]fluoroetanidazole (FETA) and now reports the oxygen dependency of binding to cells in vitro, the biodistribution of the tracer in tumor-bearing mice and the analysis of metabolites in their plasma and urine. METHODS: Four cultured rodent cell lines (V79, 36B10, EMT6 and RIF1) were incubated with [18F]FETA for various times under graded O2 concentrations. We also compared the biodistributions of [18F]FETA and [18F]fluoromisonidazole (FMISO) at 2 and 4 h postinjection in C3H mice bearing KHTn tumors (130-430 mg). Reverse-phase high-performance liquid chromatography was used to distinguish metabolites from parent drugs in urine and plasma of mice injected with [18F]FETA or [18F]FMISO. RESULTS: In cells labeled in vitro, O2 levels of 600-1300 ppm inhibited binding by 50% relative to uptake under anoxic conditions (<10 ppm). These inhibitory values are not statistically different from those reported for [18F]FMISO in the same cell lines (700-1500 ppm). In the biodistribution studies, uptake in heart, intestine, kidney and tumor was similar for both tracers 4 h after injection, whereas retention of [18F]FETA in liver and lung was significantly lower. Less uptake of [18F]FETA in liver suggests that this nitroimidazole is metabolized less than [18F]FMISO. The brain-to-blood ratios indicate that [18F]FETA readily crosses the blood-brain barrier. High performance liquid chromatography of urine demonstrated that 10% of [18F]FETA derived activity was in metabolites at 2 h postinjection, with 15% in metabolites by 4 h; comparable values for [18F]FMISO were 36% and 57%, respectively. CONCLUSION: We conclude from these data that [18F]FETA holds promise as a new hypoxia tracer in patients, having oxygen dependency of binding similar to [18F]FMISO in vitro and displaying less retention in liver and fewer metabolites in vivo. PMID- 10452327 TI - 99mTc-ENS: a new radiopharmaceutical for aerosol lung scintigraphy. Comparison between different freeze-dried formulations. AB - Exogenous natural surfactant (ENS) labeled with 99mTc(99mTc-ENS) is a new radiopharmaceutical for pulmonary aerosol scintigraphy. In this study, different freeze-dried formulations were evaluated to develop a suitable and long-storage method for the ENS, the nonradioactive precursor of this radiopharmaceutical. METHODS: Two freeze-dried formulations were evaluated: the sterile ENS suspension stannous chloride altogether lyophilized (chlorlioENS) and the lyophilized sterile ENS suspension with the addition of stannous chloride as a solid drug (lioENS). These precursors were stored at room temperature for 3 mo and then labeled with 99mTc. For comparative purposes, the sterile ENS suspension with the addition of stannous chloride labeled with 99mTc(99mTc-chlorENS) was also studied. The quality controls for each radiopharmaceutical were performed by an ascending paper chromatography to determine the labeling yield percentages. The study was performed in 30 female Sprague Dawley rats, which inhaled each radiopharmaceutical by nebulization. Twenty-five minutes after the aerosol inhalation, the animals were killed to extract their organs and measure their activity in a gamma spectrometer. The data are given as the percentage of activity concentration (C%) for each organ. RESULTS: The physicochemical properties of lioENS were adequate for a freeze-dried product. The labeling yields for 99mTc-lioENS and for 99mTc-chlorENS were always greater than 95% even after nebulization. The results of the biologic distribution studies showed that the activity concentration found in lungs for these radiopharmaceuticals were 95.7% +/- 2.6% and 96.7% +/- 2.6% respectively, results that do not differ statistically. On the other hand, the activity concentration found in lungs for the 99mTc-chlorlioENS (31.3% +/- 11.1%) and its labeling yield percentages (<10%) are statistically different (P < 0.05) from the results obtained with the two radiopharmaceuticals mentioned above. CONCLUSION: Taking into account the lioENS physicochemical properties, its long shelf life and that 99mTc-lioENS shows the same radiochemical and radiopharmacological behavior of the 99mTc-chlorENS, it can be concluded that the 99mTc-lioENS can be used for aerosol lung scintigraphy. PMID- 10452328 TI - The effect of multiple pulses of light on the circadian phase response of the rat. AB - Multiple pulses of light administered to humans have been reported to result in type 0 phase responses. These results suggest the underlying pacemaker to be nonsimple. At present, results with this type of protocol have only been reported for humans. Therefore, multiple pulses of light were administered to rats. Rats were exposed to one, two, three, or four pulses of light for 5 h (1000 lux) at successive 24-h intervals. Results did not suggest a type 0 phase response. Nonetheless, results with a second, third, or fourth light exposure were not fully predictable from a phase response curve derived from a single light pulse. PMID- 10452329 TI - Influence of pinealectomy and pineal stalk deflection on circadian gastrointestinal tract melatonin rhythms in zebra finches (Taeniopygia guttata). AB - The authors examined levels of melatonin in the plasma and various tissues in intact, pinealectomized, and pineal stalk-deflected zebra finches kept under 12:12 LD to determine if the melatonin found in the gastrointestinal tract is secreted in a circadian manner. In intact and pineal stalk-deflected birds, there is a clear day-night rhythm in melatonin content of the plasma, pineal gland, eyes, proventriculus, crop, duodenum, jejunum/ileum, colon, heart, and liver. In contrast, pinealectomy abolished the day-night rhythm. These results indicate that most of the melatonin present in the gastrointestinal tract of zebra finches is of pineal origin. However, some melatonin remained. This suggests that this melatonin may be locally synthesized and has paracrine and/or autocrine functions. Nonetheless, the results do not lend support to the contention that this putative melatonin secretion by the gastrointestinal tract is circadian. PMID- 10452330 TI - Nonentrained circadian rhythms of melatonin in submariners scheduled to an 18 hour day. AB - The human circadian timing system has previously been shown to free run with a period slightly longer than 24 h in subjects living in the laboratory under conditions of forced desynchrony. In forced desynchrony, subjects are shielded from bright light and periodic time cues and are required to live on a day length outside the range of circadian entrainment. The work schedule used for most personnel aboard American submarines is 6 h on duty alternating with 12 h off duty. This imposed 18-h cycle is too short for human circadian synchronization, especially given that there is no bright-light exposure aboard submarines. However, crew members are exposed to 24-h stimuli that could mediate synchronization, such as clocks and social contacts with personnel who are living on a 24-h schedule. The authors investigated circadian rhythms of salivary melatonin in 20 crew members during a prolonged voyage on a Trident nuclear submarine. The authors found that in crew members living on the 18-h duty cycle, the endogenous rhythm of melatonin showed an average period of 24.35 h (n = 12, SD = 0.18 h). These data indicate that social contacts and knowledge of clock time are insufficient for entrainment to a 24-h period in personnel living by an 18-h rest-activity cycle aboard a submarine. PMID- 10452331 TI - No evidence for extraocular photoreceptors in the circadian system of the Syrian hamster. AB - Campbell and Murphy reported recently that 3 h of bright light (13,000 lux) exposure to the area behind the knee caused phase shifts of the circadian rhythms of both body temperature and saliva melatonin in humans. The authors tested the hypothesis that extraocular photoreception is also involved in the circadian system of the Syrian hamster. Hamsters were bilaterally enucleated (eyes removed), and their backs were shaved. Hamsters with stable free-running rhythms in constant darkness were exposed to direct sunlight for 1 or 3 hours during their subjective night. Intact (control) animals showed phase shifts as expected, but the locomotor activity of enucleated animals was unaffected by the exposure to sunlight. The authors also measured the pineal melatonin content after exposure to sunlight. Pineal melatonin content in intact animals declined markedly as expected, but no decline was observed in the enucleated hamsters. The authors conclude that extraocular phototransduction is not capable of shifting the phase of the hamster's locomotor activity rhythm or of suppressing pineal melatonin synthesis. PMID- 10452332 TI - Light and diurnal cycle affect human heart rate: possible role for the circadian pacemaker. AB - Humans and animals demonstrate diurnal rhythms in physiology and behavior, which are generated by the circadian pacemaker, located in the supra-chiasmatic nucleus (SCN). The endogenous diurnal rhythm of the SCN is synchronized to the diurnal cycle most effectively by light. However, light also influences the SCN and its output instantaneously, as is demonstrated for the immediate effects of light on SCN neuronal firing frequency and on the output of the SCN to the pineal, inhibiting melatonin secretion. In addition to this, the circadian pacemaker modulates neuronally also other organs such as the adrenal. Therefore, the authors investigated the effect of this light input to the SCN on human heart rate, using light at different phases of the day-night cycle and light of different intensities. Resting heart rate (HR) was measured in volunteers between 20 and 40 years of age during supine, awake, resting conditions, and after 2 hours of fasting. In Experiment 1, HR was measured at different times over the day-night cycle at 0 lux and at indoor light. In Experiment 2, HR was measured at different times over the day-night cycle at controlled light intensities of 0 lux, 100 lux, and 800 lux. The authors demonstrate a clear diurnal rhythm in resting HR in complete darkness, similar to that measured under constant routine conditions. Second, it is demonstrated that light increases resting HR depending on the phase of the day-night cycle and on the intensity of light. These data strongly suggest that the circadian pacemaker modulates human HR. PMID- 10452333 TI - A model of molecular circadian clocks: multiple mechanisms for phase shifting and a requirement for strong nonlinear interactions. AB - A fundamental question in the field of circadian rhythms concerns the biochemical and molecular nature of the oscillator. There is strong evidence that circadian oscillators are cell autonomous and rely on periodic gene expression. In Drosophila, Neurospora, Aplysia, and vertebrates, circadian oscillators are thought to be based on molecular autoregulatory loops composed of transcription, translation, and negative feedback by proteins on nuclear transcription. By studying a mathematical model of molecular clocks based on this general concept, the authors sought to determine which features such clocks must have to generate robust and stable oscillations and to allow entrainment by external stimuli such as light. The model produced circadian oscillations as an emergent property even though a time delay in protein synthesis and rate constants of the feedback loop were much shorter than 24 h. Along with the delay in protein production, strong nonlinear interactions in macromolecular synthesis and nuclear feedback appeared to be required for the model to show well-behaved oscillatory behavior. Realistic phase-shifting patterns induced by external stimuli could be achieved by multiple mechanisms-namely, up- and downward perturbations of protein or mRNA synthesis or degradation rates. The model makes testable predictions about interactions between clock elements and mechanisms of entrainment and may help to understand the functions of the intricate molecular interactions governing circadian rhythmogenesis. PMID- 10452334 TI - Psychogenetically selected (Roman high- and low-avoidance) rats differ in 24-hour sleep organization. AB - A comparison of sleep organization in Roman high-(RHA/Verh) and low-(RLA/Verh) avoidance rats, which differ in the way they respond to environmental stimuli and in several neuroendocrine and neurochemical parameters, was carried out. EEG sleep recordings were obtained from adult males over 12:12 light-dark periods to determine how these two psychogenetically selected rat lines might also differ in their sleep-wake cycle. There was no significant difference in total sleep time between the two lines. However, the (hypoemotional) RHA/Verh rats showed an overall increase (percentage of total sleep) in paradoxical sleep (PS) duration, with a concomitant decrease in slow-wave sleep (SWS). During the dark phase, RHA/Verh rats showed a shorter PS latency and a larger number of PS episodes. Hourly sleep scoring also revealed a more discontinuous pattern (total sleep and PS vs. SWS) during the dark phase in RHA/Verh rats. In relation to recognized neurochemical and neuroendocrine differences between them, these rat lines may prove useful in investigations of the neurobiological mechanisms underlying sleep regulation. PMID- 10452335 TI - The endogenous melatonin profile as a marker for circadian phase position. AB - Several circadian rhythms have been used to assess the phase of the endogenous circadian pacemaker (ECP). However, when more than one marker rhythm is measured, results do not always agree. Questions then inevitably arise. Are there multiple oscillators? Are some markers more reliable than others? Masking is a problem for all marker rhythms. Masking of melatonin is minimized by sampling under dim light. The dim-light melatonin onset (DLMO) is particularly convenient since it can usually be obtained before sleep. However, assessing the DLMO in low melatonin producers may be problematic, particularly with the commonly used operationally defined threshold of 10 pg/ml. This study evaluates various circadian phase markers provided by the plasma melatonin profile in 14 individuals, several of whom are low melatonin producers. The amount (amplitude) of melatonin production appears to influence the phase of many points on the melatonin profile. Accordingly, when low producers are in a data set, we now prefer a lower DLMO threshold than the one previously recommended (10 pg/ml). Indeed, there are some low producers who never exceed this threshold at any time. Radioimmunoassays are now available that have the requisite sensitivity and specificity to support the use of a lower threshold. Nevertheless, the dim-light melatonin offset (DLMOff), even when operationally defined at thresholds less than 10 pg/ml, appears to be confounded by amplitude in this study; in such cases, it may be preferable to use the melatonin synthesis offset (SynOff) because it is not confounded by amplitude and because, theoretically, it is temporally closer to the endogenous mechanism signaling the offset of production. The question of whether the termination mechanism of melatonin synthesis is related to an interval timer or to a second oscillator loosely coupled to the onset oscillator is probably best answered using the SynOff rather than the DLMOff. It is hoped that these findings will make a useful contribution to the debate on the best ways to use points on the melatonin profile to assess circadian phase position in humans. PMID- 10452336 TI - Melatonin rhythm observed throughout a three-cycle bright-light stimulus designed to reset the human circadian pacemaker. AB - Exposure to light and darkness can rapidly induce phase shifts of the human circadian pacemaker. A type 0 phase response curve (PRC) to light that has been reported for humans was based on circadian phase data collected from constant routines performed before and after a three-cycle light stimulus, but resetting data observed throughout the entire resetting protocol have not been previously reported. Pineal melatonin secretion is governed by the hypothalamic circadian pacemaker via a well-defined neural pathway and is reportedly less subject to the masking effects of sleep and activity than body temperature. The authors reasoned that observation of the melatonin rhythm throughout the three-cycle light resetting trials could provide daily phase-resetting information, allowing a dynamic view of the resetting response of the circadian pacemaker to light. Subjects (n = 12) living in otherwise dim light (approximately 10-15 lux) were exposed to a noncritical stimulus of three cycles of bright light (approximately 9500 lux for 5 h per day) timed to phase advance or phase delay the human circadian pacemaker; control subjects (n = 11) were scheduled to the same protocols but exposed to three 5-h darkness cycles instead of light. Subjects underwent initial and final constant routine phase assessments; hourly melatonin samples and body temperature data were collected throughout the protocol. Average daily phase shifts of 1 to 3 h were observed in 11 of 12 subjects receiving the bright light, supporting predictions obtained using Kronauer's phase-amplitude model of the resetting response of the human circadian pacemaker. The melatonin rhythm in the 12th subject progressively attenuated in amplitude throughout the resetting trial, becoming undetectable for >32 hours preceding an abrupt reappearance of the rhythm at a shifted phase with a recovered amplitude. The data from control subjects who remained in dim lighting and darkness delayed on average -0.2 h per day, consistent with the daily delay expected due to the longer than 24-h intrinsic period of the human circadian pacemaker. Both temperature and melatonin rhythms shifted by equivalent amounts in both bright light-treated and control subjects (R = 0.968; p<0.0001; n = 23). Observation of the melatonin rhythm throughout a three-cycle resetting trial has provided a dynamic view of the daily phase-resetting response of the human circadian pacemaker. Taken together with the observation of strong type 0 resetting in humans in response to the same three-cycle stimulus applied at a critical phase, these data confirm the importance of considering both phase and amplitude when describing the resetting of the human circadian pacemaker by light. PMID- 10452337 TI - The Registry of the International Society for Heart and Lung Transplantation: sixteenth official report--1999. PMID- 10452338 TI - Lung allograft dysfunction correlates with gamma-interferon gene expression in bronchoalveolar lavage. AB - BACKGROUND: Preceding episodes of acute cellular rejection (ACR) may predispose lung allografts to the subsequent development of irreversible dysfunction or bronchiolitis obliterans syndrome (BOS). Other histologic patterns such as bronchiolitis obliterans with organizing pneumonia (BOOP), organizing pneumonia, lymphocytic bronchiolitis and diffuse alveolar damage (DAD) may also adversely affect allograft function. We have previously reported the predominant expression of Th1 cytokines (IL-2 and interferon gamma) in rejecting and Th2 (IL-10) in a tolerant model of rat lung transplantation. Here we correlate the "Th1/Th2 paradigm" in clinical lung transplantation with histologic findings and assess the effect on serial spirometric function. METHODS: We examined the mRNA expression of IL-2, interferon gamma, IL-10 and ICAM-1 in 53 bronchoalveolar lavage (BAL) specimens from 23 lung transplant (LT) recipients utilizing qualitative "nested" reverse transcriptase polymerase chain reaction (RT-PCR). We also measured IgG1 and IgG2 levels in 44 BAL specimens by ELISA. The mRNA expression for cytokines, ICAM-1 and the IgG2/IgG1 ratios were correlated with the presence or absence of ACR and alternate "histologic patterns". Serial spirometry were analyzed for the 2-3 month interval before bronchoscopic (FOB) assessment to derive "baseline" forced expiratory volume-one second (FEV1) values. The change in FEV1 coincident with (deltaFEV1 pre) and for the 2-3 month interval subsequent to (deltaFEV1 post) FOB were expressed relative to "baseline" spirometric indexes. RESULTS: Detection of mRNA for interferon gamma and ICAM-1 correlated significantly with ACR, whereas IL-2 and IL-10 expression did not correlate. IL-10 was virtually "ubiquitous" in most BAL samples irrespective of the presence or absence of ACR. The highest correlation was observed with interferon gamma for acute cellular rejection whereupon the sensitivity was 77.7%, specificity 87.7%, positive predictive value 73.6% and negative predictive value 88.2%, although for ICAM-1 these values were 75%, 65.7%, 50.0% and 85.0%, respectively. Nevertheless, 4 of 5 episodes of respiratory tract infection (bacterial, CMV, Aspergillus spp.) were similarly associated with cytokine mRNA. The ratios of IgG2 to IgG1, a reflection of Th1/Th2 influence, were not statistically different when analyzed for the presence or absence of ACR (0.91+/ 0.53 vs. 1.02+/-0.70, respectively; p = NS). By analysis of FEV1 trends, expression of interferon gamma was associated with a greater and persistent decrement (deltaFEV1 pre: -0.265+/-0.78 liters, and post: -0.236+/-0.1161; mean +/- SE) than ACR in the absence of interferon gamma expression (+0.158 +/- +0.065 and +0.236+/-0.007 liters, respectively) (Student-Newman-Keuls, p<.05). CONCLUSION: Our findings suggest that interferon gamma mRNA expression and ICAM-1 may be valuable in both the diagnosis and prognosis for lung allograft ACR. IL 10, a Th2 cytokine, was locally expressed both in the presence and absence of ACR. Expression of mRNA for interferon y in BAL and, to a lesser extent ICAM-1, were associated with increased lung allograft dysfunction. Whether BAL cytokine "immunosurveillance" would complement or possibly supplant a specific "histologic pattern" and thereby direct different therapies after lung transplantation, may be potentially rewarding areas of further investigation. PMID- 10452339 TI - The effects of a neutrophil elastase inhibitor (ONO-5046.Na) and neutrophil depletion using a granulotrap (G-1) column on lung reperfusion injury in dogs. AB - BACKGROUND: Activated neutrophils are reported to be closely involved in ischemia reperfusion injury after lung transplantation. We investigated the beneficial effects of a new recombinant specific neutrophil elastase inhibitor, ONO-5046.Na, and an extracorporeal-type granulotrap (G-1) column on ischemia-reperfusion lung injury, by using an in situ warm lung ischemia model in dogs. METHODS: Warm ischemia was induced for 3 hours by clamping the pulmonary arteries and veins. The left main bronchus was bisected and reanastomosed prior to reperfusion. The left lung was collapsed for 3 hours. A total of 27 adult mongrel dogs were divided into three groups: the control group (n = 9) treated with a saline vehicle; the ONO group (n = 9), in which ONO-5046.Na was continuously administrated from before induced ischemia and to ending 2 hours after reperfusion; and the G-1 group (n = 9), in which a G-1 column was applied for 90 minutes starting 30 minutes before reperfusion under passive bypass support. RESULTS: Circulating neutrophils in the G-1 group decreased significantly (p<.05) compared to preischemia, and significantly decreased compared with the other groups after reperfusion. Oxygenation was improved actually and pulmonary vascular resistance was kept lower level after the administration of ONO-5046.Na. The increase of lung weight was significantly ameliorated in both the G-1 and ONO groups. In the histopathological study, lungs from the control group demonstrated diffuse alveolar edema, neutrophil infiltration, massive alveolar exudate and hemorrhage, and thickening of the interstitium. Lungs from the G-1 group showed mild swelling of the alveolar wall and neutrophil infiltration. Lungs from the ONO group showed virtually no abnormalities. CONCLUSION: This study demonstrated that a neutrophil elastase inhibitor and neutrophil depletion prevented lung reperfusion injury. These treatments may prevent ischemia and reperfusion injury in lung transplantation. PMID- 10452340 TI - Patterns and significance of CD44 expression in lung allografts. AB - BACKGROUND: Lung allograft rejection involves the interplay of multiple cellular populations, soluble mediators, and extracellular matrix proteins. The CD44 family of cell surface glycoproteins mediates a variety of cell-cell and cell matrix interactions including lymphocyte homing to sites of antigenic challenge and fibroblast migration and invasion into extracellular matrix, processes integral to lung allograft rejection. METHODS: We performed immunohistochemical staining for CD44 on biopsies from allograft recipients with differing rejection experiences: Group 1 (n = 5 patients/10 biopsies) never exceeded Grade A1 or B2 acute rejection (AR); Group 2 (n = 7 patients/26 biopsies) had 2 or more episodes of Grade A2 or higher AR and no obliterative bronchiolitis (OB); Group 3 (n = 6 patients/17 biopsies) had clinical and pathologic OB. Nine infected allograft biopsies, 8 near-normal lung sections (non-transplant controls), and 13 non transplant biopsies showing bronchiolitis obliterans organizing pneumonia (BOOP), organizing diffuse alveolar damage (DAD), or usual interstitial pneumonia (UIP) were also studied. RESULTS: Allograft biopsies demonstrated significantly more CD44 staining among lymphocytes, macrophages, Type II pneumocytes, and respiratory epithelial cells than non-transplant controls, while staining of lymphocytes, macrophages, and Type II pneumocytes did not differ significantly between allograft groups. Fibroblast CD44 staining in Group 3 biopsies significantly exceeded that of controls and Groups 1 and 2, and biopsies with AR and/or OB showed more fibroblast staining than biopsies with BOOP, organizing DAD, or UIP. Alveolar CD44-positive fibroblasts did not predict development of OB, while bronchial CD44-positive fibroblasts were followed in one case by OB. CONCLUSIONS: These findings suggest that CD44 expression is characteristic of graft-infiltrating inflammatory cells and resident parenchymal cells, and may be related to the initiation and evolution of AR and OB. PMID- 10452341 TI - Prognostic indices in heart transplant candidates after the first hospitalization triggered by the need for intravenous pharmacologic circulatory support. AB - BACKGROUND: Patients with heart failure refractory to optimal oral pharmacologic therapy have a dismal short term prognosis. Heart transplantation is the only therapy shown to improve survival in these patients. Unfortunately, due to the critical shortage of donor organs, approximately 30% of listed patients with end stage heart failure die before a suitable donor heart becomes available. The principal aim of this study was to determine whether intravenous pharmacologic circulatory support favorably influences the clinical course of heart transplant candidates or whether mechanical circulatory support should be instituted in this high risk patient population. METHODS: Data from 154 consecutive hospitalizations in 125 patients 49+/-12 years were retrospectively reviewed. The product limit method was used to estimate survival. Multiple logistic regression analysis was used to identify the clinical and hemodynamic variables that independently predict outcome after each admission in which heart transplantation did not occur. RESULTS: One year survival for the study population was 65%. This survival is significantly lower than the 91% 1 year survival in similarly ill patients undergoing heart transplantation. The Cox proportional hazard method identified serum bilirubin, blood urea nitrogen (BUN), serum sodium levels and right atrial pressure as independent prognostic indices. Serum bilirubin, BUN levels and duration of intravenous pharmacologic circulatory support were associated with a poor outcome. A composite index including serum bilirubin and BUN levels predicted outcome with a sensitivity and specificity of 79% and 77%, respectively. The addition of pharmacologic support duration increased the model's sensitivity to 95%, but did not significantly alter specificity that was 74%. Of the 125 patients hospitalized due to the need to initiate intravenous pharmacologic support for the first time (index hospitalization), 69 (55%) were discharged after optimization of medical therapy. Of 21 patients who did not undergo transplantation during the follow-up period, 18 (86%) died within 2 years of the index hospitalization. The duration of intravenous pharmacologic support beyond which prognosis dramatically worsens without heart transplantation is 21 days. CONCLUSION: Heart transplant candidates who require intravenous pharmacologic circulatory support for more than 21 days and do not receive a suitable donor heart within this period of time have a high mortality. Alternative therapies, such as implantation of a mechanical circulatory assist device should be considered in this high risk population. PMID- 10452342 TI - Nitric oxide inhalation in the treatment of primary graft failure following heart transplantation. AB - BACKGROUND: Primary graft failure from right or left ventricular insufficiency remains a serious cause of early death following heart transplantation. Inhaled nitric oxide (NO) is a potent pulmonary vasodilator that could decrease pulmonary pressure and improve right ventricular function. METHODS: Two cases of early graft failure following orthotopic heart transplantation were treated with NO inhalation. The treatment consisted of inhalation of 20 ppm of NO, introduced 4 to 6 hours following transplantation, in 2 patients supported with high doses of inotropic agents and vasopressors in addition to the intra-aortic balloon pump. RESULTS: In the first and second cases, NO inhalation resulted in a decrease in pulmonary artery pressure, in a decrease in pulmonary vascular resistance and in an increase in cardiac index. In the second patient, systemic oxygenation improved markedly 30 minutes after initiation of NO. In the 2 patients, NO inhalation, mechanical ventilation and the intra-aortic balloon pump were weaned 4 days following transplantation. CONCLUSION: Primary graft failure from donor ischemic damage, reperfusion injury or pulmonary hypertension remains a serious complication. The use of an intra-aortic balloon pump, inotropic agents and of inhaled NO appears to offer the best support for recovery of donor heart function. Primary graft failure from right or left ventricular insufficiency remains a serious cause of early mortality following heart transplantation. Ischemic damage of donor heart, reperfusion injury or pulmonary hypertension are the main causes of early graft failure. Although the cause is multifactorial, treatment of primary organ failure remains difficult with dismal results. The objective of the present study was to review the result of 2 patients with donor right heart failure following heart transplantation treated with inhaled nitric oxide (NO). PMID- 10452343 TI - An analysis of the effect of age on survival after heart transplant. AB - BACKGROUND: Advances in immunosuppression and reports of improved survival after cardiac transplantation have led to a liberalization of traditional recipient eligibility criteria, especially age. While age alone is not a contraindication to transplantation, conflicting data exists regarding long-term survival of the older transplant recipient. METHODS: One hundred-fifty three patients undergoing consecutive first time cardiac transplantation from June 7, 1985 through February 1, 1997 were studied. For purposes of analysis, patients were stratified according to age (<55 years vs. >55 years) and hospital and late outcomes determined. RESULTS: The incidence of early and late acute cellular rejection was not different based up on age. The freedom from infection at 12 months was 54+/ 5% for patients < or =55 compared to 32+/-8% for patients >55 years old (p = .04). Five year estimated survival for patients >55 years old was only 56+/-9% compared to 78+/-5% for patients < or =55 years old (p = .005). The hazard for death was highest within the first post-transplant year for older patients and was most commonly due to infection. Both advanced age and pre-transplant diagnosis of ischemic cardiomyopathy were found to be independently and additively predictive of reduced late survival. CONCLUSIONS: In the present study, late survival was adversely influenced by advanced age. Older patients (>55 years) with pre-transplant diagnosis of ischemic cardiomyopathy were particularly at high risk (risk ratio 4.6:1) for death. Given little prospect of expanding the number of donor hearts, careful selection of patients over the age of 55 with pre-transplant ischemic cardiomyopathy is warranted. PMID- 10452344 TI - Extracellular calcium concentration affects susceptibility to global ischemic injury in newborn but not adult hearts. AB - BACKGROUND: Whether immaturity in calcium handling, that persists for a time after birth, could increase sensitivity to extracellular calcium and affect the development of global ischemic injury in the newborn heart is unknown. To address this, the impact of alterations in extracellular calcium concentration on newborn vs. adult development of myocardial injury due to ischemia was studied. METHODS: In Study 1, hearts of 3-day-old piglets and adult pigs were perfused with 1 of 3 different calcium concentrations: control (0.13 mmol/L); intermediate (2.23 mmol/L); high (4.44 mmol/L) before normothermic ischemia. In Study 2, newborn hearts were allocated to perfusion with or without the L-calcium channel antagonist verapamil before high (4.44 mmol/L) calcium exposure, followed by normothermic ischemia. Tolerance to ischemia was assessed by determining the time to irreversible injury in all hearts, and maximal intraventricular pressures at peak injury. RESULTS: In adults, altering calcium did not significantly affect tolerance to ischemia. In newborns, increasing calcium exposure resulted in significantly greater intraventricular pressures at maximal injury when compared with the control (low) calcium group (p<.05). As well, newborns exposed to high calcium had a significantly shorter time to the development of ischemic injury compared with the other groups (p<.05). Those newborn hearts pretreated with an L calcium channel antagonist before the high calcium exposure did not exhibit this increased susceptibility to ischemic injury (p<.05). CONCLUSIONS: In contrast to adults, the development of ischemic injury in the newborn heart is affected by changes in extracellular calcium, that can be modified with an L-calcium channel antagonist. This information could be used to prolong the safe preservation time of newborn donor hearts harvested for transplantation, as well as to minimize postoperative ventricular dysfunction. PMID- 10452345 TI - Combined use of prostacyclin and higher perfusate temperatures further enhances the superior lung preservation by Celsior solution in the isolated rat lung. AB - BACKGROUND: The poor tolerance of the lung to ischemia and reperfusion (IR) still represents one of the limitations in clinically successful lung transplantation. Modified Euro-Collins (EC) is routinely used in lung preservation, but alternative solutions have been developed for improvement of pulmonary preservation. Celsior is an extracellular solution that has significantly reduced the IR-induced pulmonary damage in animal studies. So far, no extensive experimental studies exist concerning the influence of Celsior on pulmonary gas exchange following IR. METHODS: In an extracorporeal rat lung model 10 lungs, each, were preserved with Celsior (CE) and Celsior/prostacyclin (CEPC, 6 microg/100 ml) at 4 degrees and 15 degrees C, each, and compared to low-potassium Euro-Collins (EC-40, 40 mmol/liter potassium). After 2 hours of ischemia lungs were reventilated and reperfused using a roller pump. Oxygenation in terms of oxygen partial tension in the left atrial effluent, pulmonary vascular resistance (PVR), peak inspiratory pressure, and wet/dry ratio were monitored for 50 minutes. Furthermore, edema formation was evaluated by light microscopy. Statistical analysis was performed using ANOVA models. RESULTS: Compared to the EC-40 group, oxygenation was increased and amount of edema was reduced in most Celsior-preserved organs (p<0.032) with exception of the CEPC group at 4 degrees C (p = 0.06). Additional application of prostacyclin did not have any significant effect on oxygenation in the Celsior group. However, after temperature elevation of the CEPC perfusate to 15 degrees C, a superior partial tension of oxygen was observed (p<0.023) in contrast to the 4 degrees C groups CE and CEPC. The lowest PVR was found in the CE 4 degrees C group (p<0.02). CONCLUSIONS: Celsior provides better lung preservation than EC-40 solution. Application of prostacyclin at higher perfusate temperatures results in additional functional improvement. In vivo experiments and ultrastructural analysis are warranted for further evaluation of Celsior in lung preservation. PMID- 10452346 TI - Relation between long-term steroid treatment after heart transplantation, hypofibrinolysis and myocardial microthrombi generation. AB - BACKGROUND: Thrombotic complications and transplant coronary artery disease are among the main causes of morbidity and mortality after heart transplantation. A thrombophilic state has been described in transplant recipients, and correlated to immunosuppressive therapy with cyclosporine A or azathioprine, whereas the prothrombotic effects of steroids, even though always given, have never been duly considered. A reduced fibrinolytic capacity due to high levels of PAI-1, the most important inhibitor of plasminogen activators, was suggested to play a role in the development of cardiovascular diseases and transplant coronary artery disease. A severe hypofibrinolytic state secondary to PAI-1 increase has been found in patients with Cushing's disease, and in hypercorticism secondary to long term steroid treatment after renal transplantation. METHODS: We evaluated plasma clotting and fibrinolytic behaviors in 2 groups of heart transplant patients treated with (26 cases) or without (23 cases) steroids together with cyclosporine A and azathioprine. Twenty-five healthy subjects were studied as normal controls. The following tests were assayed at least 1 year after transplantation: fibrinogen, factor VIII coagulant activity, von Willebrand factor antigen, euglobulin lysis time, tissue plasminogen activator antigen and activity, PAI-1 antigen and activity. In addition, the presence of cardiac microthrombi was evaluated on 2 endomyocardial biopsy specimens obtained in each patient both on day 7 after heart transplantation (first control) and usually 1 year or more later (last control). RESULTS: Plasma levels of fibrinogen, factor VIII and von Willebrand factor were significantly higher in both groups of patients than in normal controls. Fibrinolytic activity was significantly reduced in transplant patients treated with steroids, compared with steroid-free patients and normal controls. In steroid-treated heart transplant recipients, the hypofibrinolytic state was due to a significant and pathological increase in PAI-1 antigen and activity levels. The fibrinolytic impairment was more evident in patients transplanted for ischemic heart disease and treated with steroids than in patients with previous dilated cardiomyopathy and treated either with or without steroids. Myocardial microthrombi were found in 2/49 cases at the first biopsy control, and in 12/49 cases at the last biopsy control after transplantation. This different prevalence was statistically significant (chi2 = 8.33, p = .003). Plasma PAI-1 activity was significantly higher and, as a consequence, euglobulin lysis time was more prolonged in microthrombi-positive patients than in microthrombi-negative ones. Among the 12 transplant recipients who developed cardiac microthrombi, 7 patients were treated with steroids and showed higher PAI 1 levels and more reduced fibrinolytic activity than the 5 steroid-free patients. CONCLUSIONS: Our data confirm the prothrombotic state induced by long-term steroid treatment, characterized by an increase in PAI-1 levels and secondary impairment of fibrinolytic capacity. In heart transplant patients, steroid related hypofibrinolysis might constitute a further risk factor for transplant coronary artery disease. PMID- 10452347 TI - Plasmapheresis with intravenous immunoglobulin G is effective in patients with elevated panel reactive antibody prior to cardiac transplantation. AB - BACKGROUND: Patients with a PRA >10% are considered to be at greater risk for the development of not only acute cellular and humoral rejection but also increased mortality when compared to nonsensitized patients following transplantation. All patients with a PRA >10% at our institution are treated with plasmapheresis and intravenous immunoglobulin G immediately prior to cardiac transplantation. METHODS: Sixteen (Group 1) of 118 patients awaiting cardiac transplantation were found to be sensitized. These patients underwent plasmapheresis followed by 20 gm of intravenous immunoglobulin G (IVIG) immediately prior to cardiac transplantation. Group 1 was compared to the remaining 102 patients with a PRA <10% (Group 2). RESULTS: Despite more patients in Group 1 having a positive crossmatch, pulmonary hypertension, and requiring mechanical circulatory support, there was no statistically significant difference in length of stay or mortality at a mean follow-up of 21.6+/-15.0 months. There was no difference in the occurrence of mild, moderate or severe cellular rejection or humoral rejection in these sensitized patients when compared to Group 2. CONCLUSIONS: Pretransplant plasmapheresis followed by intravenous immunoglobulin G may be an effective therapy that obviates the need for a prospective crossmatch and allows sensitized patients to undergo cardiac transplantation. There is no increase in the post transplant length of stay, occurrence of rejection or short term mortality. Long term follow up is necessary to evaluate whether there is a difference in the development of late rejection, transplant vasculopathy and survival. PMID- 10452348 TI - Tacrolimus-based triple-drug immunosuppression minimizes serum lipid elevations in pediatric cardiac transplant recipients. AB - BACKGROUND: Immunosuppression with corticosteroids and cyclosporine has been associated with hyperlipidemia, a risk factor for post-transplant coronary artery disease. The recent development of tacrolimus has created an alternative to cyclosporine-based triple drug immunotherapy. One potential benefit that has been reported in patients receiving tacrolimus is a minimization of elevation of both total and LDL cholesterol, compared to those increases observed in patients receiving cyclosporine-based immunosuppression. It is unclear in previous studies whether this beneficial effect is related to tacrolimus directly or to its corticosteroid sparing potential. To study this relationship, we compared lipid profiles from pediatric cardiac transplant recipients treated with corticosteroids, and either cyclosporine or tacrolimus. METHODS: The study group consisted of 23 patients (mean age = 12.3 years) with pre-transplant and serial post-transplant determinations of total cholesterol, LDL, HDL, and triglycerides. Patients were separated into 4 study groups, defined by immunosuppressive regimen (cyclosporine vs. tacrolimus) and prednisone dose (>0.10 mg/kg/day vs. < or =0.10 mg/kg/day). RESULTS: Patients who received cyclosporine and higher doses of prednisone experienced a mean 74 mg/dl increase from baseline in total cholesterol (p = .0001). None of the other 3 treatment groups demonstrated a statistically significant elevation. Similar trends were observed in LDL and triglyceride alterations between the 4 study groups. Interestingly, patients treated with tacrolimus and higher doses of prednisone demonstrated a significant rise in HDL from baseline (p = .0001), although those who received cyclosporine and higher dose prednisone failed to exhibit this rise. CONCLUSION: The minimal degree of lipid alteration seen in patients receiving tacrolimus and higher doses of prednisone indicates that this effect was not solely based upon the steroid sparing properties of tacrolimus therapy. The data also suggests a possible synergistic effect between cyclosporine and higher doses of prednisone on hyperlipidemia. Therefore, in pediatric patients requiring higher corticosteroid doses late after transplantation, use of tacrolimus rather than cyclosporine may lead to more favorable lipid profiles and help minimize the risk of post transplant coronary arteriopathy. PMID- 10452349 TI - The histology of subcutaneously implanted donor bronchial rings correlates with rejection scores of lung allografts in a primate lung transplant model. AB - BACKGROUND: The diagnosis of acute rejection in lung transplantation generally relies on transbronchial biopsies. This invasive procedure may be associated with bronchial bleeding or pneumothorax and may not be feasible in patients with severely compromised lung function. The hypothesis of the current study was that histopathological findings of donor bronchial segments implanted into the subcutaneous tissue of lung allograft recipients would predict lung tissue rejection scores, thus providing the clinician with an alternate source of information. METHODS: Unilateral left lung transplantation was performed in 34 cynomolgus monkeys as part of a drug efficacy study. After completion of the transplant procedure, 4 bronchial ring segments of the explanted recipient left lung and 4 bronchial ring segments of the non-transplanted right donor lung were implanted subcutaneously in the abdominal region. Lung allograft rejection was evaluated by open lung biopsies of the allograft performed on postoperative (PO) Day 14 and during sacrifice on PO Day 28. At the time of each biopsy, 2 donor and 2 recipient subcutaneous bronchial rings were explanted. Histologic evaluation of the lung tissue samples was performed according to the working formulation of the International Society for Heart and Lung Transplantation. Bronchial rings were independently evaluated by assessing the degree of airway narrowing; percentage of intact epithelial coverage as well as its specific histology (respiratory ciliated, flattened cuboidal, squamous); presence of lymphocytes, macrophages or spindle cells; and presence of peribronchial inflammation, luminal fibrosis, lymphocytic bronchitis or luminal mucous. Statistical analysis was performed by logistic regression. RESULTS: In the recipient bronchial rings, there was no evidence of airway narrowing. There was 98% epithelial coverage, 71% that were respiratory ciliated cells, and there was no inflammation. Donor bronchial rings showed no airway narrowing for monkeys with grade A0 to A2 rejection in tissue biopsies and a maximum narrowing (41.2%) with A4 rejection. Epithelial cell coverage was approximately 100% with grade A0-A2 and 44+/-11% with A4 rejection. Lymphocytic bronchitis was most severe in A4 rejection and minimal in A0 to A2 rejection. By logistic regression analysis, independent predictors of a likelihood of rejection were the degree of airway obliteration, the percentage of epithelial cell coverage, the degree of lymphocytic bronchitis and the product of respiratory and flattened cuboidal cell coverage. CONCLUSIONS: The current data show that histologic alterations of subcutaneously implanted donor bronchial rings correlate with lung tissue biopsy scores based on the ISHLT working formulation. Because subcutaneous bronchial rings can be explanted under local anesthesia, they may provide useful information for the diagnosis of acute allograft rejection in patients with impaired lung function, patients that obtaining lung tissue samples may not be feasible. PMID- 10452350 TI - Dobutamine stress echocardiography: experience in pediatric heart transplant recipients. AB - BACKGROUND: Transplant coronary arteriopathy causes late death and is difficult to detect noninvasively. Dobutamine stress echocardiography is being used for risk stratification in adult recipients at some transplant centers, thus we investigated its role in a pediatric population. METHODS: We performed 46 stress echo studies (mean age = 11.8 years; mean years post transplantation = 4.3). An atropine/dobutamine protocol (5-40 mcg/kg/min) was used to attain a predicted target heart rate. Serial echocardiographic images were acquired at baseline and at each increment of dobutamine and recovery, and were digitized online. Data were correlated with endomyocardial biopsy (n = 23), coronary angiography (n = 26) or autopsy (n = 6). All studies were well tolerated. RESULTS: Target heart rate was achieved in 41/46 (89%) studies. The mean heart rate significantly increased from 95 to 169 beats/min and mean systolic blood pressure from 123 to 153 mm Hg (p<.05). The mean peak pressure-rate product was 23,041 beats-mm Hg/min. Coronary arteriopathy was confirmed in 5 patients by angiography (n = 3) explanted heart (n = 1) or autopsy (n = 4). In this group, abnormalities included a new reversible wall motion abnormality (n = 2), left ventricular cavity dilation with stress (n = 3), ischemia (n = 2), increased mitral insufficiency (n = 1) and marked diastolic dysfunction (n = 1). A positive study predicted death or graft failure (p< .0005). CONCLUSIONS: Echocardiographic abnormalities during stress correlated with coronary arteriopathy in this small cohort of patients; however, larger multi-center studies are warranted to assess the utility of dobutamine stress echocardiography for risk stratification for coronary disease in pediatric transplant recipients. PMID- 10452351 TI - The history of radial glia. AB - Radial glial cells are now recognized as a transient population that serves as scaffolding for neuronal migration. The recognition of the existence and role of radial glia has not been smooth, and here we provide a brief historical overview on the pioneering studies on this subject. The histologists and embryologists Albert Kolliker and Wilhelm His performed seminal investigations on cortical morphogenesis in the last decades of the 19th century. However, the introduction of the silver impregnation Golgi technique, and its diffusion in the late 1880s, played a crucial role in the detection of radial glial processes. The radial arrangement of fibers emerging from the neuroepithelium lining the central canal was initially detected in the embryonic spinal cord by Camillo Golgi himself. The first Golgi impregnation of the cerebral cortex of mammalian fetuses was performed by Giuseppe Magini, who detected radial fibers extending from the ventricular neuroepithelium, and observed cells intercalated along these processes. Radial fibers, regarded as epithelial or ependymal processes, were then observed in the developing spinal cord and cerebral cortex by several investigators. Santiago Ramon y Cajal was the first to suggest that radial fibers were modified astrocytic processes functioning as a support during cortical histogenesis. Cajal acknowledged Magini's findings, but he criticized Magini's observations on the existence of neurons along radial fibers. With the advent of electron microscopy, the existence of radially arranged glial processes along which young neurons migrate was finally ascertained in the early 1970s by Pasko Rakic, thus opening a new era in the cellular and molecular biology of radial glia. PMID- 10452352 TI - Imidazoline receptors: from discovery to antihypertensive therapy (facts and doubts). AB - The hypothesis and indirect evidence of imidazoline receptors has been promoted since some 15 years ago and it gave a substantial impetus for research in this field, resulting in a better understanding of neuronal and cardiovascular regulatory processes. The nomenclature of the imidazoline receptors has been accepted by international forums but no direct proof for the existence of these receptors has been published. Authors summarise the most important available data, including facts and doubts as far as the discovery, characterisation, and function of imidazoline receptors and their subtypes, the differences between imidazoline receptors and alpha-2 adrenoceptors, and also on their participation in regulatory processes. PMID- 10452353 TI - Extensive neuronal cell death following intracranial transection of the facial nerve in the adult rat. AB - The aim of the present study is to examine the neuronal degeneration and the glial response following intracranial transection of the facial nerve close to the brainstem and furthermore to compare the results with a distal nerve injury. The facial nerve was cut either intracranially in the posterior cranial fossa or further distally, where it passes the parotid gland, in adult rats. Intracranial axotomy caused a massive loss of neuronal profiles. Only 26.8+/-11.3% of facial motor neuronal profiles were found ipsilateral to the nerve injury when compared to the contralateral side, following intracranial axotomy. This was statistically significant in comparison to the distal injury (72.4+/-9.5%), 4 weeks post lesion. Reactive microglial cells expressed ED1 immunoreactivity following the intracranial axotomy but not following the distal nerve injury. In conclusion, there was a large discrepancy in neuronal degeneration as well as presence of phagocytic (ED1 positive) microglia between the two lesions. The intracranial lesion model used in the present study generates a massive neuronal cell death and should therefore be a useful tool for studies on proximal cranial nerve injuries and in particular mechanisms causing cell death, which may occur following, for example, head trauma. PMID- 10452354 TI - [Arg8]-vasopressin-induced increase in intracellular Ca2+ concentration in cultured rat hippocampal neurons. AB - Changes in intracellular Ca2+ concentration ([Ca2+]i) induced by [Arg8] vasopressin (AVP) were studied in cultured rat hippocampal neurons by fura-2 fluorometry. AVP (10-1,000 nM) caused a dose-dependent increase in [Ca2+]i. The selective V1 vasopressin receptor agonist [Phe2, Ile3, Orn8]-vasopressin also induced a significant increase in [Ca2+]i, whereas the selective V2 vasopressin receptor agonist [deamino Cys1, D-Arg8]-vasopressin showed no effect. The AVP induced increase in [Ca2+]i was inhibited by the selective V1 vasopressin receptor antagonist d(CH2)5[Tyr2(Me), Arg8]-vasopressin and nonpeptide V1 antagonist OPC-21268. On the other hand, no antagonistic effects were observed with the V2 vasopressin antagonist desglycinamide-[d(CH2)5, D-Ile2, Ile4, Arg8] vasopressin and nonpeptide V2 antagonist OPC-31260. The increase in [Ca2+]i induced by AVP was abolished after removal of extracellular Ca2+. In addition, AVP-induced [Ca2+]i elevation was not affected by treatment with verapamil, which blocked the [Ca2+]i increase induced by an isotonic high K(+)-medium (50 mM). However, omega-conotoxin GVIA completely inhibited the effect of AVP. These results suggested that the AVP-induced [Ca2+]i increase in cultured rat hippocampal neurons is due to influx of Ca2+ through V1 VP receptors coupled with N-type calcium channels. PMID- 10452355 TI - Regional variations in spermine levels in the developing rat brain following exposure to lead. AB - Polyamines are ubiquitous compounds involved in growth and differentiation, and omithine decarboxylase (ODC), is their rate-limiting enzyme. In an effort to examine whether changes in ODC enzyme activity are reflected in alterations in the availability of polyamines, levels of the more stable end-product spermine, were examined following exposure to lead (Pb). Rats were lactationally exposed to 0.2% Pb-acetate from birth to weaning. At postnatal days 3, 5, 10, 15, and 20, pups were sacrificed, and spermine was extracted from their brain tissue and the extracts were analyzed by high-performance liquid chromatography coupled with a fluorescence detector. Basal spermine levels in the cerebellum were found to be higher than those in the neocortex, and exhibited a characteristic developmental profile. In Pb-exposed animals, spermine levels were attenuated in both brain regions, however, there was a rebound in cerebellar levels during the third week after birth. While basal spermine levels are in concert with reported patterns of ODC activity, they depart from each other following a toxic challenge to the cerebellum. This study suggest that developmental polyamine levels are not necessarily a direct translation of ODC activity and that modulations in their biosynthesis depend on regional stages of growth. PMID- 10452356 TI - Localization of the Norrie disease gene mRNA by in situ hybridization. AB - Norrie disease is a rare X-linked recessive neurodevelopmental disorder. The affected males manifest congenital blindness, which is often associated with hearing loss, mental retardation and psychiatric problems. Genetic linkage studies have localized the gene to the short arm of the X-chromosome and the gene has been isolated recently. The encoded protein is a member of the superfamily of growth factors containing a cystine knot motif and may be involved in cell adhesion and neurodevelopment. Molecular genetic analysis revealed a large number of missense, nonsense, deletion, and splice-site mutations among Norrie patients. In order to further determine the role of the Norrie disease gene, we studied the distribution pattern of its mRNA in the retina and in brain by in situ hybridization. The results show abundant hybridization signals in outer nuclear, inner nuclear, and ganglion cell layers of the retina in all three species (mice, rabbit, and human) examined. There was no significant expression in the vitreous body, lens, and rod outer segment. High expression levels were also observed in the cerebellar granular layer, hippocampus, olfactory bulb, cortex, and epithelium of the rabbit brain. These data suggest that the Norrie disease gene could play a critical role in the differentiation or maintenance of the differentiated state of the retina. PMID- 10452357 TI - Prenatal hypoxia-ischemia alters expression and activity of nitric oxide synthase in the young rat brain and causes learning deficits. AB - Inhibition of nitric oxide synthase (NOS) is known to possibly impair learning and memory. Our previous studies have demonstrated that prenatal hypoxia-ischemia (HI) decreases NOS expression and NOS activity in the neonatal rat brain. To investigate whether effects of prenatal HI on NOS expression continue and whether prenatal HI affects learning and memory in young rats, NOS expression and NOS activity were determined in the hippocampus of rat brains at 28 days of age following a prenatal HI insult on G17. Performances in the passive avoidance test and the Morris water maze test were also studied in these young rats prior to sampling. Rat fetuses were subjected to either a 30-min prenatal HI insult or a sham operation (SH) on gestation day 17 and rat pups were delivered naturally. Increased locomotor activity was observed in the prenatal HI rats as compared to the SH rats on postnatal days 13 and 15, but not on postnatal days 20 and 30. Prenatal HI affected learning ability in these young rats at 28 days of age, as indicated by a delayed acquisition of passive avoidance and by longer escape latency in the Morris water maze test as compared to the SH group. Prenatal HI did not affect retention of passive avoidance and spatial memory. Concomitant with these learning deficits, expression of neuronal NOS and endothelial NOS mRNAs as well as Ca2(+)-dependent NOS activity in the hippocampus of the prenatal HI rat brain were significantly decreased as compared to the SH group. These results suggest that a 30-min prenatal HI insult on gestation day 17 in rats has long-lasting effects on NOS expression and NOS activity in the offspring brain and on learning ability of these young rats. The learning deficit in offspring is possibly associated with the reduction in expression of NOS mRNA and NOS activity in the hippocampus of these animals. PMID- 10452359 TI - Neuronal life and death decisions functional antagonism between the Trk and p75 neurotrophin receptors. PMID- 10452358 TI - Discordant xenografts: different outcome after mouse and rat neural tissue transplantation to guinea-pigs. AB - Embryonic neural tissue obtained from other species has been considered as a donor tissue source in repair strategies for human neurodegenerative disorders. The neuro- and immunobiology of distantly related species combinations, discordant xenografts, need to be characterised. For this purpose, a small animal model would be an important research tool. Adult guinea-pigs, and adult rats as controls, received intrastriatal grafts of either mouse or rat embryonic ventral mesencephalic tissue. The survival rates and types of host immune response were assessed at 2 weeks after grafting using stereological techniques and semi quantitative evaluations. In the mouse-to-guinea-pig group, all transplants were rejected and no tyrosine hydroxylase-immuno reactive (TH-IR) cells remained. In the rat-to-guinea-pig group, there was good survival of TH-IR cells (5050 SEM+/ 1550), similar to that in the rat-to-rat group (4900 SEM+/-1540). In the mouse-to rat group, half of the animals had no surviving TH-IR cells (520 SEM+/-230 for the whole group). These species combinations offer inexpensive, efficient, and suitable conditions to study important survival factors for discordant xenogeneic neural tissue transplants. The factors responsible for the divergent graft outcomes between the two combinations might provide clues on how to manipulate xenogeneic tissue to increase survival rates in the future. PMID- 10452360 TI - Ca2+ mobilization and capacitative Ca2+ entry regulate DNA synthesis in cultured chick retinal neuroepithelial cells. AB - Release of Ca2+ from intracellular Ca2+ stores (Ca2+ mobilization) and capacitative Ca2+ entry have been shown to be inducible in neuroepithelial cells of the early embryonic chick retina. Both types of Ca2+ responses decline parallel with retinal progenitor cell proliferation. To investigate their potential role in the regulation of neuroepithelial cell proliferation, we studied the effects of 2,5-di-tert-butylhydroquinone (DBHQ), an inhibitor of the Ca2+ pump of intracellular Ca2+ stores, and of SK&F 96365, an inhibitor of capacitative Ca2+ entry, on DNA synthesis in retinal organ cultures from embryonic day 3 (E3) chicks and in dissociated cultures from E7 and E9 chick retinae. We demonstrate that both antagonists inhibit [3H]-thymidine incorporation in a dose-dependent manner without affecting cell viability or morphology. The inhibition of [3H]-thymidine incorporation by SK&F 96365 occurred in the same concentration range (IC50: approximately 4 microM) as the blockade of capacitative Ca2+ entry in the E3 retinal organ culture. At a concentration of 5 microM SK&F 96365. DNA synthesis was reduced by 71, 40 and 32% in the E3, E7 and E9 cultures, respectively. Application of DBHQ at concentrations which led to depletion of intracellular Ca2+ stores also inhibited [3H]-thymidine incorporation with IC50 values of 20-30 microM in the different cultures. Our results suggest the involvement of Ca2+ mobilization and capacitative Ca2+ entry in the regulation of DNA synthesis in the developing neural retina. PMID- 10452361 TI - Developmental regulation of glutamate transporters and glutamine synthetase activity in astrocyte cultures differentiated in vitro. AB - Glutamate plays an important role in brain development, physiological function, and neurodegeneration. Astrocytes control synaptic concentration of glutamate via the high affinity glutamate transporters, GLT-1 and GLAST, and the glutamate catabolizing enzyme, glutamine synthetase. In this study we show that astrocytes cultured from rat brain in various stages of development including embryonic (E18), postnatal (P1-P21) and mature (P50), show distinct patterns of GLT-1 and GLAST expression, glutamine synthetase activity, and phenotypic changes induced by dibutyryl-cyclic adenosine monophosphate. The transcripts for GLT-1 message were detectable in embryonic astrocytes only, whereas the GLAST message was highly expressed in E18 and P1-P4 astrocyte cultures, declined in P10-P21, and was undetectable in P50 astrocytes. Uptake of 3H-glutamate correlated well with GLAST expression in astrocyte cultures of all developmental stages. Glutamine synthetase activity significantly declined from high embryonic levels in P4 astrocytes and remained low throughout postnatal maturation. Exposure of astrocyte cultures to the differentiating agent, db-cAMP (250-500 microM; 6 days), resulted in a pronounced stellation, up-regulation of GLT-1 and GLAST in E18, and GLAST in P4 cultures, while it was ineffective in P10 astrocytes. By contrast, db-cAMP induced a more pronounced stimulation of glutamine synthetase activity (up to 10-fold above basal) in P10 than in E18 cultures (up to 2 times above basal). The differences in expression/inducibility of glutamate transporters and glutamine synthetase observed in astrocyte cultures derived from various stages of fetal and postnatal development suggest that astrocytes in vivo might also respond differently to environmental or injurious stimuli during development and maturation. PMID- 10452362 TI - Neuron loss in the mouse hippocampus following prenatal injection of tritiated thymidine or saline. AB - To investigate possible effects of injections of tritiated thymidine ([3H]dThd) into pregnant mice or the injection procedure itself on the proliferation of neuronal precursor cells in the fetuses, pregnant mice received intraperitoneal injections of either [3H]dThd or saline on embryonic days 12, 14, and 19, while their offspring remained untreated. A second group of dams was not injected but their male offspring received a subcutaneous injection of again either [3H]dThd or saline on postnatal day 10. Then total numbers of hippocampal pyramidal cells (areas CA1 to CA3) and granular cells (dentate gyrus) were determined stereologically for 20-day-old as well as for 80-day-old male pups. No significant differences were found for the mean total number of pyramidal cells between the investigated groups of pups. However, the mean total number of granular cells was significantly reduced in those groups in which the dams had received an intraperitoneal injection, irrespective of whether [3H]dThd or saline was injected. This revives the repeated warning in the literature to consider the effect of the injection procedure on the developing brain when interpreting possible effects of agents administered during pregnancy. PMID- 10452363 TI - FGF-9 is an autocrine/paracrine neurotrophic substance for spinal motoneurons. AB - Motoneurons need muscle-derived neurotrophic substances for their survival during the initial phase of their development, but after maturation they lose this requirement and can survive after axotomy. This suggests that some neurotrophic substances other than target-derived ones control the survival of motoneurons in adults. Because spinal motoneurons express fibroblast growth factor-9 (FGF-9) messenger RNA, we hypothesized that FGF-9 might be an autocrine or paracrine survival factor for motoneurons. FGF-9 promoted the survival of motoneurons and upregulated the choline acetyl-transferase (ChAT) activity in the dissociated cultures of ventral half of rat E13 spinal cord. Externally added FGF-9 was more effective in low density cultures, and polyclonal blocking antibody against FGF-9 significantly lowered the ChAT activity. Our results support an autocrine or paracrine role for FGF-9 in mediating the survival of spinal motoneurons. Non target-derived neurotrophic substances for motoneurons including FGF-9 should be important in the pathogenesis of motor neuron disorders in the adults, especially amyotrophic lateral sclerosis. PMID- 10452364 TI - GABAergic system in the developing mammalian retina: dual sources of GABA at early stages of postnatal development. AB - In the present work, we have characterized the maturation of the GABAergic system in mammalian retina. Immunoreactivity for GABA, GAD (glutamic acid decarboxylase, EC 4.1.1.15) -65 and -67 in the adult rat retina was localized in cells in the inner nuclear and ganglion cell layers. This pattern was established around postnatal day 8 and included transient GABA and GAD-67 expression in horizontal cells. GAD activity was very low at P1 and P4, increasing after P8, reaching maximal activity by P21 and decreasing to attain adult values by P30. GABA content was approximately constant from P1 to P13, increasing thereafter to reach adult levels. GAD protein content increased progressively with postnatal development and the two isoforms could be distinguished at P8. The disparity between retinal GABA content vs. presence and activity of the synthesizing enzyme, led us to investigate the alternative pathway for GABA synthesis that utilizes putrescine as a substrate. Highest levels of ornithine decarboxylase activity (the limiting step for putrescine synthesis) were found between P1 and P4, decreasing to very low levels after P13. The same pattern was observed for putrescine content in the retina. Highest amounts were found at P1, that decreased and remained constant after P13. Additionally, approximately 40% of tritiated putrescine incorporated by P1, P4 and adult retinas was converted into GABA. Our results suggest the existence of two different sources of GABA in mammalian retina, one that uses glutamate as a precursor and predominates in the mature nervous system and another that utilizes putrescine and is present transiently at early developmental stages. PMID- 10452365 TI - Spatiotemporal pattern of the postnatal astrogliogenesis in the rat hippocampal formation. AB - Newborn, 2-, 4-, 8-, 16- and 30-day-old Wistar rats were injected with 3H thymidine and sacrificed following 4 h. survival time. Brain sections containing the dorsal part of hippocampal formation were immunostained for S-100beta protein and subjected to autoradiography to visualize proliferating astrocytes. Microscopical observations revealed age-dependent changes in the number and distribution of proliferating astrocytes. The changes were considered as being related to the neurogenetic gradient characteristic to the hippocampal formation. PMID- 10452366 TI - Horizontal cell differentiation in the retina of the Brazilian opossum, Monodelphis domestica. AB - Differentiation of many diverse neuronal phenotypes is an essential part of nervous system development. We have studied the differentiation of horizontal cells, one of the basic neuronal types in the vertebrate retina, in a small, easily maintained marsupial by immunocytochemistry using antineurofilament and antivimentin antibodies. At birth the retina consists of proliferating neural epithelial cells, with a few early ganglion cells. Horizontal cells were first detected in 12-day-old pups; somas were within the epithelial neuroblastic layer and processes extended radially. By 19 days there were tangentially oriented dendrites and a few longer processes, the beginning of the outer plexiform (first synaptic) layer. By the time of eye opening (about 34 days) the basic histological organization of the mature retina was established. In the mature retina and during development, horizontal cell neurites in the outer plexiform layer, as well as ganglion cell axons, reacted strongly with several antineurofilament antibodies and with antivimentin; horizontal cell somas were detected only with one antineurofilament antibody. Only one population was detected, which we identify as the short-axon subtype, by comparison with horizontal cells in other marsupials and in eutherian mammals. This is the first description of the putative absence in a marsupial of one of the two horizontal cell subtypes found in most amniotes, including mammals so far studied, except murid rodents, which have only the short-axon subtype. Absence of one subtype in Monodelphis supports the hypothesis that the short-axon cell is the basic conserved phenotype of this class and suggests that experimental analysis of differentiation of horizontal cells in Monodelphis and murid rodents, compared to marsupials and eutherian mammals which have the basic two subtypes, can help elucidate mechanisms for controlling differentiation of specific cellular phenotypes and the variations in neurons within and among species. PMID- 10452367 TI - Quantitative study of plasticity in the auditory nuclei of chick under conditions of prenatal sound attenuation and overstimulation with species specific and music sound stimuli. AB - Morphological effects of prenatal sound attenuation and sound overstimulation by species specific and music sounds on the brainstem auditory nuclei of chick have been evaluated quantitatively. Changes in length, volume, neuron number, size of neuronal nuclei and glial numbers of second and third order auditory nuclei, n. magnocellularis (NM) and n. laminaris (NL), were determined from thionine-stained serial sections of control and experimental groups on posthatch day 1 using stereological methods. Significant increase in volume of both auditory nuclei attributable to increase in length of nucleus, number and size of neurons, number of glia as well as neuropil was observed in response to both species specific and music overstimulation given during the critical period of development. The enhanced development of auditory nuclei in response to enriched environment prenatally indicates a positive effect of activity on neurons which may have clinical implications in addition to providing explanation for preference to auditory cues in the postnatal life. Reduction in neuron number with a small increase in proportion of cell nuclei of large size as well as an increase in glial numbers was seen in both NM and NL of the prenatally sound attenuated chicks. The increase in size of some neuronal nuclei may probably be evidence of enhanced synthesis of proteins involved in cell death or an attempt at recovery. The dissociated response of neurons and glia under sound attenuated and auditory stimulated conditions suggests that they are independently regulated by activity dependent signals with glia also being under influence of other signals for a role in removal of dead cell debris. PMID- 10452368 TI - Carbon filaments direct the growth of postlesional plastic axons after spinal cord injury. AB - The effect of implantation of carbon filaments and fetal tissues on the axonal regeneration following contusion injury in a rat model was investigated by in situ immunofluorescence. Female Sprague-Dawley rats were subjected to severe contusion injury to the spinal cord at T9-T10. All animals were divided into 5 groups (N = 5/group): normal controls. surgical controls, with carbon filament implants, with fetal tissue implants and with implants consisting of fetal tissue cocultured with carbon filaments. After a 10-week survival period, the astroglial response was assessed by immunoreactive glial fibrillary acidic protein and the neuro-axonal profile by immunoreactive phosphorylated and nonphosphorylated neurofilament proteins. The contusion injury resulted in: (a) dramatically increased immunoreactivity of glial fibrillary acidic protein indicating injury associated reactive astrogliosis, (b) increase in immunoreactive phosphorylated neurofilament protein indicating upregulated phosphorylation of neurofilament protein, (c) with no change in the highly differentiated nonphosphorylated neurofilament protein which normally occur in the nonregenerating mature neurons. Implantation of fetal tissues alone following contusion injury did not show any appreciable change with regard to the immunoreactivities for the glial and neuronal markers studied, compared to the injury controls. However, the implantation of carbon filaments alone or together with fetal tissues directed the growth of glial fibrillary acidic protein-positive astroglia and phosphoneurofilament-positive neurites along the carbon fibers, with no effect on nonphosphoneurofilament protein. In conclusion, implantation of carbon filaments appears to be critical for facilitating the attachment of astroglia forming a substrate and scaffolding that can further support and direct the growth of postlesional plastic axons across the lesion. In addition, carbon filament prostheses in combination with fetal tissue implants provides an improved combinational approach to promote regrowth of injured neurons following injury. PMID- 10452369 TI - Social separation in infant Cebus apella: patterns of behavioral and cortisol response. AB - 34 infant tufted capuchins (Cebus apella) were separated from their social groups for a 2-hour period and videotaped in isolation at the ages of 6 months and 1 year. Baseline and 2-hour blood samples were measured for levels of serum cortisol. Compared to homecage baseline levels, passivity, locomotion and vocalizations increased during separation, while self-directed behavior and environmental exploration decreased. Both behavioral and cortisol responses to separation showed individual stability over the 6 month period, although both responses were somewhat attenuated at the later age. There was little correlation between cortisol and behavior during separations. Females vocalized more than did males during separations and showed greater cortisol increases at 6 months of age. The pattern of behavioral response seen in the 2 hours following separation appeared to be more passive than the typical 'protest' response described in many nonhuman primates, and may reflect either the physical circumstances of the separation or a characteristic of species with relaxed social bonds and considerable allomothering available to infants. PMID- 10452370 TI - Gene transfer into the mammalian inner ear using HSV-1 and vaccinia virus vectors. AB - The introduction of foreign genes into cells has become an effective means of achieving intracellular expression of foreign proteins, both for therapeutic purposes and for experimental manipulation. Gene delivery to the nervous system has been extensively studied, primarily using viral vectors. However, to date less work has focused on gene delivery to the inner ear, and existing studies have primarily used adenovirus and adeno-associated virus. Using two recombinant viral vectors, herpes simplex type 1 (HSV-1), and vaccinia virus, bearing the Escherichia coli lacZ gene, we tested gene delivery to the guinea pig cochlea in vivo with beta-galactosidase staining as an assay. The HSV-1 and vaccinia virus vectors were both found to infect and elicit transgene expression successfully in many cells in the guinea pig cochlea, including cells in the organ of Corti. These data demonstrate the feasibility of gene delivery to the inner ear using these two viral vectors. Such techniques may facilitate study of the auditory systems, and might be used to develop gene therapy strategies for some forms of hearing loss. PMID- 10452371 TI - Quantitative measure of genetic differences in susceptibility to noise-induced hearing loss in two strains of mice. AB - The CBA/CaJ (CB) and C57BL/6J (B6) inbred strains of mice were exposed for 1 h to noise intensities between 98 and 119 dB SPL. Previous studies indicated that the B6 mice exhibited permanent threshold shifts (PTS) after 1h exposure to 110 dB, whereas the CB mice did not exhibit any PTS. These differences in susceptibility to noise-induced hearing loss (NIHL) appear to be due to a gene for age-related hearing loss (AHL). The current study was designed to determine dose-response curves for NIHL over the ranges of intensities of noise that would characterize the B6 and CB inbred strains of mice. Because of the considerable differences in sensitivity to NIHL, the noise exposures for the two strains overlapped only at 110 and 113 dB. Nevertheless, the two strains exhibited two different dose response curves, offset and with different slopes. We postulate that the B6 strain of mice exhibits a more linear increase for PTS from 98-113 dB, consistent with incremental effects on some metabolic physiological mechanism(s); the abrupt transition in NIHL between 113 and 116 dB for the CB mice is consistent with an ototraumatic structural injury. PMID- 10452373 TI - Distortion product otoacoustic emissions in the CBA/J mouse model of presbycusis. AB - CBA mice do not exhibit age-related loss of auditory sensitivity or cochlear pathology until relatively late in life. Therefore, this strain is believed to be an excellent animal model for the examination of the effects of age on the cochlea. To evaluate the effects of age on outer hair cell function, 2f1-f2 distortion product otoacoustic emissions (DPOAEs) were measured for f2 between 8 and 16 kHz in CBA/J mice between 1 and 25 months of age. CBA mice exhibited mild age-related changes in DPOAE level and detection threshold at 17 months of age, and changes of 20-40 dB by 25 months of age. The DPOAE level decreased and detection threshold increased with age in a frequency-dependent manner, starting at high frequencies and eventually extending to low frequencies. The range of frequencies in which notches were observed in the DPOAE input/output (I/O) functions extended toward lower frequencies by 17 months of age. Notches were absent in the I/O functions of 25-month-old mice. The present results for a frequency range of 8-16 kHz suggest that age has modest effects on outer hair cell function in CBA mice. PMID- 10452372 TI - Distributed representation of spectral and temporal information in rat primary auditory cortex. AB - Modulations of amplitude and frequency are common features of natural sounds, and are prominent in behaviorally important communication sounds. The mammalian auditory cortex is known to contain representations of these important stimulus parameters. This study describes the distributed representations of tone frequency and modulation rate in the rat primary auditory cortex (A1). Detailed maps of auditory cortex responses to single tones and tone trains were constructed from recordings from 50-60 microelectrode penetrations introduced into each hemisphere. Recorded data demonstrated that the cortex uses a distributed coding strategy to represent both spectral and temporal information in the rat, as in other species. Just as spectral information is encoded in the firing patterns of neurons tuned to different frequencies, temporal information appears to be encoded using a set of filters covering a range of behaviorally important repetition rates. Although the average A1 repetition rate transfer function (RRTF) was low-pass with a sharp drop-off in evoked spikes per tone above 9 pulses per second (pps), individual RRTFs exhibited significant structure between 4 and 10 pps, including substantial facilitation or depression to tones presented at specific rates. No organized topography of these temporal filters could be determined. PMID- 10452374 TI - Age-related sensitivity to cisplatin ototoxicity in gerbils. AB - This study was undertaken to define the developmental period of maximal sensitivity to cisplatin ototoxicity in gerbils. Five groups were established based upon post-natal age (P) at exposure to cisplatin, P10 (n = 8), P14 (n = 8), P18 (n = 6), P22 (n = 7) and P42 (n = 7). Animals were given cisplatin, 1 mg/kg/day intraperitoneal for 4 days. In the first four groups, P10, P14, P18 and P22, distortion product otoacoustic emissions were measured at 45 days of age, when responses were expected to be developmentally stable. Distortion product grams and input-output functions were measured. There was a statistically significant difference only between P14 and P42 (P<0.01). There was a significant interaction of age and frequency in the P14 group only (P<0.01). A secondary analysis compared distortion product grams of P14 animals, exposed to cisplatin, and age-matched saline-treated animals (n = 6). There was a significant treatment effect. In summary, there was an effect of age on the cisplatin ototoxicity in gerbils. Also, there was an effect of the frequency on DPOAE levels in P14 gerbils. These data support the presence of a 'sensitive' period to cisplatin ototoxicity in gerbils. PMID- 10452375 TI - K+ and Na+ absorption by outer sulcus epithelial cells. AB - Transduction of sound into nerve impulses by hair cells depends on modulation of a current carried primarily by K+ into the cell across apical transduction channels that are permeable to cations. The cochlear function thus depends on active secretion of K+ accompanied by absorption of Na+ by epithelial cells enclosing the cochlear duct. The para-sensory cells which participate in the absorption of Na+ (down to the uniquely low level of 1 mM) were previously unidentified and the existence of a para-sensory pathway which actively absorbs K+ was previously unknown. A relative short circuit current (Isc,probe, measured as the extracellular current density with a vibrating electrode) was directed into the apical side of the outer sulcus epithelium, decreased by ouabain (1 mM), an inhibitor of Na+, K(+)-ATPase, and found to depend on bath Na+ and K+ but on neither Ca2+ nor Cl-. Isc,probe was shown to be an active current by its sensitivity to ouabain. On-cell patch clamp recordings of the apical membrane of outer sulcus cells displayed a channel activity, which carried inward currents under conditions identical to those used to measure Isc,probe. Both Isc,probe and non-selective cation channels (27.4+/-0.6 ps, n = 22) in excised outside-out patches from the apical membrane were inhibited by Gd3+ (1 mM). Ics,prob was also inhibited by 5 mM lidocaine, 1 mM quinine and 500 microM amiloride but not by 10 microM amiloride. These results demonstrate that outer sulcus epithelial cells contribute to the homeostasis of endolymph by actively absorbing Na+ and K+. An entry pathway in the apical membrane was shown to be through non-selective cation channels that were sensitive to Gd3+. PMID- 10452376 TI - Olivocochlear neurons in the chinchilla: a retrograde fluorescent labelling study. AB - Although the chinchilla is widely used as a model for auditory research, little is known about the distribution and morphology of its olivocochlear neurons. Here, we report on the olivocochlear neurons projecting to one cochlea, as determined by single and double retrograde fluorescent tracer techniques. 10 adult chinchillas were anesthetized and given either unilateral or bilateral injections of a fluorescent tracer (either Fluoro-Gold or Fast Blue) into scala tympani or as a control, a unilateral injection into the middle ear cavity. The results indicate that there are similarities as well as significant differences between the chinchilla and other species of rodents in the distributions of their olivocochlear neurons. Based on three well-labelled cases, there was a mean total of 1168 olivocochlear neurons in the chinchilla. Of these, the majority (mean 787) were small, lateral olivocochlear neurons found almost exclusively within the ipsilateral lateral superior olivary nucleus. The next largest group consisted of a mean of 280 medial olivocochlear neurons virtually all of which were located in the dorsomedial peri-olivary nucleus. Chinchilla medial olivocochlear neurons were more predominantly crossed in their projections (4:1) than in any known species. The smallest group of olivocochlear neurons (mean 101) consisted of larger lateral olivocochlear neurons (shell neurons) which were located on the margins of the superior olivary nucleus and which projected mainly (2.2:1) ipsilaterally. Double retrograde labelling was observed only in medial olivocochlear neurons and occurred in only 1-2% of these cells. The results confirm previous findings which indicated a relative paucity of fibers belonging to the uncrossed as compared to the crossed olivocochlear bundle. This, together with the strong apical bias of the uncrossed projection reported previously, offers possible explanations for the apparent absence of efferent-mediated suppressive effects of contralateral acoustic stimulation in this species. Regarding the lateral olivocochlear system, the chinchilla is shown to possess both intrinsic and shell neurons, as in the rat. PMID- 10452377 TI - Effects of a hair cell transcription factor, Brn-3.1, gene deletion on homozygous and heterozygous mouse cochleas in adulthood and aging. AB - The transcription factor Brn-3.1, is expressed in the inner ear hair cells throughout life and is necessary for the development of these cells. Mutant mice in which the Brn-3.1 encoding region has been deleted have no identifiable hair cells, greatly reduced numbers of spiral ganglion cells and are deaf. A mutation in the human homologue of this gene has been shown to be related to adult onset, sensorineural hearing loss (Vahava et al., 1998). The question whether haploinsufficiency in the mutant Brn-3.1 mouse with a mixed C57BL6/129Sv genetic background could affect the adult or aged cochlea was tested, therefore, by measuring the auditory brainstem responses and examining the cochlea's histologically at 2, 18 and 24 months of age. The heterozygotes had a comparable hearing to the wild-type animals and similar patterns of cochlear degeneration. Both groups showed an about 30 dB hearing loss beginning at 18 months of age, outer hair cell degeneration and loss of spiral ganglion neurons in the basal turn. There appeared to be no effect of Brn-3.1 haploinsufficiency on the mouse cochlea, implying that one intact copy of the gene is sufficient to maintain a normal cochlea. PMID- 10452378 TI - Gamma-aminobutyric acidergic and glycinergic inputs shape coding of amplitude modulation in the chinchilla cochlear nucleus. AB - Amplitude modulation is a prominent acoustic feature of biologically relevant sounds, such as speech and animal vocalizations. Enhanced temporal coding of amplitude modulation signals is found in certain dorsal and posteroventral cochlear nucleus neurons when they are compared to auditory nerve. Although mechanisms underlying this improved temporal selectivity are not known, involvement of inhibition has been suggested. gamma-Aminobutyric acid- and glycine-mediated inhibition have been shown to shape the dorsal cochlear nucleus and posteroventral cochlear nucleus response properties to other acoustic stimuli. In the present study, responses to amplitude modulation tones were obtained from chinchilla dorsal cochlear nucleus and posteroventral cochlear nucleus neurons. The amplitude modulation carrier was set to the neuron's characteristic frequency and the modulating frequency varied from 10 Hz. Rate and temporal modulation transfer functions were compared across neurons. Bandpass temporal modulation transfer functions were observed in 74% of the neurons studied. Most cochlear nucleus neurons (90%) displayed flat or lowpass rate modulation transfer functions to amplitude modulation signals presented at 2540 dB (re: characteristic frequency threshold). The role of inhibition in shaping responses to amplitude modulation stimuli was examined using iontophoretic application of glycine or gamma-aminobutyric acidA receptor agonists and antagonists. Blockade of gamma-aminobutyric acidA or glycine receptors increased stimulus-evoked discharge rates for a majority of neurons tested. Synchronization to the envelope was reduced, particularly at low and middle modulating frequencies, with temporal modulation transfer functions becoming flattened and less bandpass in appearance. Application of glycine, gamma-aminobutyric acid or muscimol increased the modulation gain over the low- and mid-modulation frequencies and reduced the discharge rate across envelope frequencies for most neurons tested. These findings support the hypothesis that glycinergic and gamma aminobutyric acidergic inputs onto certain dorsal cochlear nucleus and posteroventral cochlear nucleus neurons play a role in shaping responses to amplitude modulation stimuli and may be responsible for the reported preservation of amplitude modulation temporal coding in dorsal cochlear nucleus and posteroventral cochlear nucleus neurons at high stimulus intensities or in background noise. PMID- 10452379 TI - Across-species comparisons of psychophysical detection thresholds for electrical stimulation of the cochlea: I. Sinusoidal stimuli. AB - Several species have been, and continue to be, used as subjects in studies of electrical stimulation of the cochlea. Few attempts, however, have been made to determine if data obtained from different species are quantitatively or qualitatively similar. The present work compares psychophysical absolute detection threshold vs. frequency functions for sinusoidal stimuli obtained from humans, nonhuman primates, cats, and guinea pigs. Threshold data for monopolar and bipolar electrode configurations from both previously published and unpublished studies are compared. In general, within all four species, significant intersubject variation in detection threshold level was found, but slopes of threshold vs. frequency functions were relatively well conserved within a species, under the conditions studied. With one exception (cat bipolar stimulation), threshold functions reached a minimum at or near 100 Hz across species and electrode configurations. In all cases, thresholds were significantly lower for monopolar, as compared with bipolar, configurations. Statistically, there were no significant differences in absolute threshold level across species. Threshold levels increased with frequency above 100 Hz at a rate of 3.0-7.9 dB/octave, depending on both electrode configuration and species. Slopes were steeper for monopolar than for bipolar configurations. When slopes were averaged between 200 and 2000 Hz, no statistically significant differences in overall slopes were found, nor was there a significant interaction between electrode configuration and species. There were, however, consistent species differences within more restricted regions of the function. Human functions for both monopolar and bipolar stimulation were steeper than all animal functions in the range of 100-300 Hz. Within this range, the differences between slopes for human and nonhuman subjects were statistically significant. In addition, differences were noted in the frequency at which slope decreased, with slopes for nonhuman subjects showing the decrease at higher frequencies than did those for human subjects. These differences may be true species differences, or may reflect the influence of confounding variables associated with each experimental-subject model. PMID- 10452380 TI - Effects of stimulus level on electrode-place discrimination in human subjects with cochlear implants. AB - Effects of stimulus level on discrimination of one stimulation site from another were examined in 15 human subjects with Nucleus-22 cochlear implant systems. Bipolar stimulation was used in all cases with electrodes in the bipolar pair separated by 1.5 mm (center to center). Subjects were first tested at a medium loudness level, using an adaptive tracking procedure, to determine the regions of the electrode array where electrode-place discrimination was best and the regions where it was poorest. Electrode-place discrimination was then tested at three regions distributed throughout the array, which included the regions of best and poorest discrimination. At each region, electrode-place discrimination was tested at three levels: 25%, 50%, and 75% of the dynamic range. For each of these nine conditions (3 sites x 3 levels), the test-electrode pairs were loudness balanced with the reference-electrode pairs. A two-interval forced-choice same-different procedure was then used to determine discriminability of the reference-electrode pair from the nearest, apical, test-electrode pair. If P(C)max was <0.707 at all three levels, additional testing was done using the next, more apical, electrode pair as the test-electrode pair. A tendency toward better discrimination at more apical regions of the array was observed. Electrode pairs with poor discrimination typically had smaller dynamic ranges than those with good discrimination. There was a weak tendency toward better discrimination at higher levels of stimulation. However, effects of level on electrode-place discrimination were less pronounced and less consistent than previously observed effects of level on temporal discriminations. These results suggest interactions between current spread and the condition of the implanted cochlea as underlying mechanisms. PMID- 10452381 TI - Chromosomal localization of a gene responsible for vestibulocochlear defects of BUS/Idr mice: identification as an allele of waltzer. AB - Mice of the bustling mutant strain BUS/Idr have vestibulocochlear defects. bus/bus homozygotes, but not heterozygotes, are hyperactive and display an abnormal behavior such as circling, head bobbing and head tilting. To characterize BUS mice further, the auditory brain-stem response of the mutant was examined. In +/bus heterozygotes as well as control animals, the auditory brain stem response was developmentally first recorded as early as 11 days of age and heterozygous and normal adults showed typical auditory brain-stem responses with five peaks in a threshold of 40-45 dB SPL. In contrast, bus/bus homozygotes showed no auditory brain-stem response at any age in response to stimuli up to 130 dB SPL, indicating that they are deaf throughout life. Linkage analysis revealed that the responsible gene, originally designated as bus, maps on chromosome 10, 1.09+/-0.9 cM distal to D10Mit127 and D10Mit59, and 0.72+/-0.51 cM proximal to three markers, D10Mit48, D10Mit112 and D10Mit258, at a site indistinguishable from that of the Albany waltzer, v(A/b). The results of allelism tests between BUS and Albany waltzer indicated that bus is allelic with v(Alb). From these data, we propose here that the bus mutation could represent another allele of waltzer, now designated v(bus). PMID- 10452382 TI - Spatiotemporal representation of binaural difference in time and intensity of sound in the guinea pig auditory cortex. AB - Neural activity of the auditory cortex (AC) in response to a change of interaural intensity difference (IID) and interaural time difference (ITD) of sound stimuli was observed by optical recording with a 12 x 12 photodiode array and the voltage sensitive dye, RH795. Guinea pigs (280-450 g) were anesthetized with sodium pentobarbital (30 mg/kg) and supplemental doses of neuroleptic solutions. When both ears were stimulated dichotically by tone bursts (14 kHz, 75 dB SPL), excitatory optical signals appeared in both anterior (A) and dorsocaudal (DC) fields of AC. An increase of intensity of ipsilateral stimulation from 65 to 95 dB SPL caused a decrease of neural activity of isofrequency bands in both fields. An increase of ipsilateral leads from -2.5 to 10 ms resulted in a gradual decrease of the amplitude of the excitatory responses. A strong inhibition was observed in field DC and the ventral portion of field A. These results show the different spatiotemporal representation of IID and ITD sensitivities in AC. However, the ipsilateral lead inducing a large inhibition was much longer than the time difference (80 micros) calculated from the interaural distance of the guinea pig. This indicates that the longer binaural inhibition observed in AC would have a different functional significance from that of the neural system of ITD detection in the guinea pig. PMID- 10452383 TI - Positron emission tomography of cortical centers of tinnitus. AB - Tinnitus is associated with a wide variety of disorders in the auditory system. Whether generated peripherally or centrally, tinnitus is believed to be associated with activity in specific cortical regions. The present study tested the hypothesis that these cortical centers subserve the generation, perception and processing of the tinnitus stimulus and that these processes are suppressed by lidocaine and masking. Positron emission tomography was used to map the tinnitus-specific central activity. By subtracting positron emission tomography images of regional cerebral blood flow distribution obtained during suppression of the tinnitus from positron emission tomography images obtained during the habitual tinnitus sensation, we were able to identify brain areas concerned with the cerebral representation of tinnitus. Increased neuronal activity caused by tinnitus occurred predominantly in the right hemisphere with significant foci in the middle frontal and middle temporal gyri, in addition to lateral and mesial posterior sites. The results are consistent with the hypothesis that the sensation of tinnitus is associated with activity in cortical regions functionally linked to subserve attention, emotion and memory. For the first time, the functional anatomy of conditions with and without the habitual tinnitus sensation was obtained and compared in the same subjects. PMID- 10452384 TI - The effects of quinine on cochlear nerve fibre activity in the guinea pig. AB - The effect of quinine on single cochlear nerve fibre activity (n = 38) was measured in four pigmented guinea pigs, which were given 10-30 mg/kg of quinine intravenously. The frequency tuning curves of these fibres exhibited significant increases in the thresholds of both 'tip' and 'tails' regions of the frequency tuning curve, but these changes did not appear to be accompanied by significant changes in tuning, as measured by the Q'10'dB. In comparison with control fibres (n = 178) from 13 untreated animals, significant changes in the proportion of low:high spontaneous rates (SR) were also seen. Using a boundary criterion of 25 sp/s, this rate changed from 26:74% to 47:53% in control and quinine-poisoned fibres, respectively. Independent of changes in the spontaneous rate, significant increases in the mean absolute refractory period from 0.85 to 1 ms were measured following quinine administration. The absence of a significant effect on fibre tuning whilst threshold was elevated indicates that quinine does not affect the integrity of the cochlear amplifier, though appears to affect cochlear sensitivity. PMID- 10452385 TI - Development of human cochlear active mechanism asymmetry: involvement of the medial olivocochlear system? AB - To study the functional development of the medial olivocochlear system, transient evoked otoacoustic emission suppression experiments were conducted in 73 ears of 38 pre-term and 11 full-term neonates. The continuous contralateral stimulation was a broad band white noise, presented at 70 dB SPL. Efferent suppression was determined by subtracting the without-contralateral stimulation condition from the with-contralateral stimulation condition. Across this population, a mean suppression effect of contralateral stimulation on transient-evoked otoacoustic emissions was found, with most of the suppression effect observed after 8 ms. The amount of suppression is linearly, positively correlated with the conceptional age. In the subgroup of bilaterally tested neonates, the suppression of transient evoked otoacoustic emissions is similar in the right ear and the left ear in subjects whose conceptional age is less than 36 weeks and significantly higher in the right ear than in the left ear in older neonates. This last observation was seen at frequencies where transient-evoked otoacoustic emission amplitudes became higher in the right ear than in the left ear as the conceptional age increased, a finding already reported in adults. This study shows that the functional adult pattern of the medial efferent system, probably involved in the detection of signals in noise such as speech sounds, seems to appear gradually in neonates and represents one of the several arguments in favor of functional auditory lateralization in humans, with a right ear advantage. PMID- 10452386 TI - Survival-fixation of the cochlea: a technique for following time-dependent degeneration and repair in noise-exposed chinchillas. AB - To minimize problems with data interpretation due to interanimal variation in susceptibility to noise, we developed a survival-fixation paradigm which involves fixing one cochlea of an experimental chinchilla at one post-exposure time and fixing the second cochlea as much as 14-24 days later. This paradigm is analytically effective because there is a high correlation in the magnitude and pattern of damage in the left and right cochleas of binaurally exposed animals. Thus, each experimental animal provides two snapshots in the degeneration and repair continua in which it can be certain that both cochleas sustained equivalent amounts of damage during the exposure. Using this technique, the time course of degeneration of different structures and cells in the organ of Corti can be determined and primary damage can be distinguished from secondary effects. The present paper discusses the issues which had to be addressed to develop this technique and provides preliminary results from chinchillas exposed to a traumatic noise. PMID- 10452388 TI - Government advertises key FSA posts. Food Standards Agency. PMID- 10452387 TI - Micromechanical effects in the cochlea of tetracaine. AB - Local anesthetics applied in the tympanic cavity have earlier been shown to affect the gross receptor potentials in reducing the cochlear microphonics and increasing the positive summating potential. To study the effects of this drug on the mechanical responses in the cochlea, vibrations were measured using laser heterodyne interferometry in an isolated in vitro temporal bone preparation from the guinea pig. Measurements were made at a set of frequencies in the fourth cochlear turn from the Hensen's cells and the outer hair cells in response to sound applied to the ear. The tuning curves of the fundamental and the second harmonic components of the vibratory responses were plotted. When 2 mM tetracaine was applied, the high frequency slope of the second harmonic curve shifted down in frequency, this caused the frequency of the maximum of second harmonic tuning to shift down. These changes were reversible when tetracaine was washed out. Observations were also made in the temporal bone preparation in vitro with a confocal microscope. Fluorescent probes were used to label various structures in the organ of Corti. Optical sections were obtained by tilting the organ permitting a view from the side like a radial section through the organ. Images were acquired before, during and after application of tetracaine and were later analyzed with a computer program. Simultaneously, cochlear microphonics and the summating potential were obtained to monitor the electrical response of the preparation. Although the cochlear microphonics and summating potential decreased when 2 mM tetracaine was applied, structural changes were not measurable in the organ of Corti. The decrease was reversible when tetracaine was washed out. It is concluded that tetracaine affected the high frequency part of the non-linear second harmonic component, possibly by lowering the stiffness of the stereocilia bundle or the body of the outer hair cells. PMID- 10452389 TI - Neospora caninum continues to be implicated in bovine abortion cases. PMID- 10452390 TI - Orf (contagious pustular dermatitis) in farmworkers: prevalence and risk factors in three areas of England. AB - Orf is a zoonotic skin disease which is commonly self-diagnosed by those who tend sheep and goats. This paper reports the prevalence, incidence and risk factors associated with the infection in a cohort of farmworkers from three areas of England, derived from the results of self-reporting and serology. Twenty-three per cent of those employed or living on a sheep farm reported ever having had orf, and the antibody serological profiles indicated a prevalence of 4 per cent and an annual incidence of 2.8 per cen. The main risk factors associated with the infection were contact with sheep, the size of the sheep flock, and contact with dogs. PMID- 10452391 TI - Clinical and radiographic findings compared with urodynamic findings in neutered female dogs with refractory urinary incontinence. AB - Possible correlations were investigated between the clinical and urodynamic findings in 77 neutered female dogs with urinary incontinence refractory to standard medical management. The clinical data used in the analysis were age, bodyweight, breed, the characteristics of the dribbling of urine and whether it developed before or after neutering, the type of response to previous treatments, signs of polyuria/polydipsia, and the results of urinalysis and culture. In 50 of the dogs, a standardised contrast radiographic examination was made to locate the neck of the bladder and to assess the morphology of the bladder. These findings were analysed for their correlation with the results of multichannel urethral pressure profilometry and diuresis cysto-urethrometry. Logistic regression analysis revealed that neither the clinical nor the radiographic findings could predict any of the abnormal urodynamic findings. Functional abnormalities of the bladder, such as a low compliance, detrusor instability, or a low bladder capacity may therefore be particularly difficult to detect in dogs with urinary incontinence. PMID- 10452392 TI - Sudden death of two horses associated with pulmonary aspergillosis. AB - The sudden death of two horses was attributed to the rapid and acute development of pulmonary aspergillosis. One horse was making excellent postoperative progress after a jejunal resection and anastomosis for intestinal adhesions. The other horse was being treated routinely for equine protozoal myeloencephalitis (EPM). Signs of fever and an increased respiratory rate were detected shortly before death in the first horse, but no premonitory clinical signs characteristic of pulmonary infection were detected in the horse being treated for EPM. Both horses developed rapidly debilitating, acute pulmonary mycosis and died unexpectedly. PMID- 10452393 TI - Isolation of ortho- and paramyxoviruses from wild birds in Northern Ireland during the 1997 Newcastle disease epizootic. PMID- 10452394 TI - Estimation of the prevalence of severe combined immunodeficiency disease in UK Arab horses as determined by a DNA-based test. PMID- 10452395 TI - Mutational analysis of the tumour suppressor gene p53 in lymphosarcoma in two bull mastiffs. PMID- 10452396 TI - Tuberculosis in cattle and badgers. PMID- 10452397 TI - Annual warble fly survey, 1998/99. PMID- 10452398 TI - Chemical destruction of horses. PMID- 10452399 TI - Withdrawal of Immobilon LA. PMID- 10452400 TI - The farming crisis. PMID- 10452401 TI - Do giraffe roar? PMID- 10452402 TI - Balance of the sexes. PMID- 10452403 TI - Balance of the sexes. PMID- 10452404 TI - Guidelines for sugar consumption in Europe: is a quantitative approach justified? AB - OBJECTIVES: There is incongruity between the sugar consumption guidelines set in different European countries. A number have adopted maximum limits ranging from 10-25% energy, while others have no quantitative recommendations at all. This raises the question whether or not there should be a common European guideline for sugar consumption. DESIGN: This paper examines if such a goal for sugar is merited and reviews the published literature on associations between sugar consumption and dental caries, obesity and micronutrient dilution. RESULTS: Evidence showed that higher intakes of sugar were related to leanness, not obesity, and had no detrimental effects on micronutrient intakes in most people. In the case of dental caries, there was a relationship between frequency of sugar intake and the incidence of decay. However, in populations where fluoride use was adequate, associations between sugar intake and caries rarely reached statistical significance. CONCLUSIONS: The available evidence does not justify a common quantitative recommendation for sugar. It is suggested that dental caries merits a more integrated public health approach where advice on the frequency of foods containing fermentable-carbohydrates is placed in context alongside oral hygiene. PMID- 10452405 TI - Exercise training and blood lipids in hyperlipidemic and normolipidemic adults: a meta-analysis of randomized, controlled trials. AB - OBJECTIVE: To determine the effectiveness of exercise training (aerobic and resistance) in modifying blood lipids, and to determine the most effective training programme with regard to duration, intensity and frequency for optimizing the blood lipid profile. DESIGN: Trials were identified by a systematic search of Medline, Embase, Science Citation Index (SCI), published reviews and the references of relevant trials. The inclusion criteria were limited to randomized, controlled trials of aerobic and resistance exercise training which were conducted over a minimum of four weeks and involved measurement of one or more of the following: total cholesterol (TC), high density lipoprotein (HIDL-C), low density lipoprotein (LDL-C) and triglycerides (TG). SUBJECTS: A total of 31 trials ( 1833 hyperlipidemic and normolipidemic participants) were included. RESULTS: Aerobic exercise training resulted in small but statistically significant decreases of 0.10 mmol/L (95% CI: 0.02, 0.18). 0.10 (95% CI: 0.02, 0.19), 0.08 mmol/L (95% CI: 0.02, 0.14), for TC, LDL-C, and TG, respectively, with an increase in HDL-C of 0.05 mmol/L (95% CI: 0.02, 0.08). Comparisons between the intensities of the aerobic exercise programmes produced inconsistent results; but more frequent exercise did not appear to result in greater improvements to the lipid profile than exercise three times per week. The evidence for the effect of resistance exercise training was inconclusive. CONCLUSIONS: Caution is required when drawing firm conclusions from this study given the significant heterogeneity with comparisons. However, the results appear to indicate that aerobic exercise training produced small but favourable modifications to blood lipids in previously sedentary adults. PMID- 10452406 TI - Dietary intakes of polyunsaturated fatty acids and indices of oxidative stress in human volunteers. AB - OBJECTIVE: To assess whether nutritionally-relevant changes in polyunsaturated fatty acid (PUFA) intake alter indices of oxidative stress in human volunteers DESIGN: A split plot/change over dietary study where half the volunteers consumed a diet containing 5% PUFA (low PUFA) as food energy for 4 weeks and after a 6 week washout period consumed a 15% PUFA (high PUFA) diet for another 4 weeks. The second group of volunteers completed this protocol in reverse. Total fat, carbohydrate, protein and vitamin E contents of the diets were constant. SUBJECTS: 10 healthy, non-smoking, male volunteers aged 32.6 +/- 1.7 y RESULTS: There was a significant increase in whole blood oxidised glutathione (P < 0.05), an index of oxidative stress, after consumption of the high PUFA diet. Moreover, urinary thiobarbituric acid reactive substances (TBARS), an index of lipid peroxidation, significantly increased (P = 0.038) following consumption of the high PUFA diet and decreased (P = 0.031) after consuming the low PUFA diet. However, there was no change in non specific plasma indices of lipid peroxidation, conjugated dienes and TBARS, nor in red cell antioxidant enzymes glutathione peroxidase, glutathione reductase, and catalase. However, superoxide dismutase significantly decreased (13%, P=0.018) after consumption of the low PUFA diet. Total cholesterol increased by 13% (P=0.014) after consumption of the low PUFA diet. CONCLUSIONS: This study indicates that although increasing dietary levels of PUFA may favourably alter cholesterol profiles, the same dietary changes may adversely affect some indices of lipid peroxidation. Care should be taken when providing dietary advice on PUFA intake and an adequate intake of antioxidants to match any increased PUFA may be important for preventing oxidative stress. PMID- 10452407 TI - Impact of zinc supplementation on subsequent growth and morbidity in Bangladeshi children with acute diarrhoea. AB - OBJECTIVE: To assess the impact of zinc supplementation during acute diarrhoea on subsequent growth and morbidity in malnourished young children. DESIGN: Double blind randomized controlled clinical trial SETTING: International Centre for Diarrhoeal Disease Research, Bangladesh. SUBJECTS: Sixty-five children aged 3-24 months with acute diarrhoea for less than 3 d. INTERVENTION: Either elemental zinc (20 mg/d) in a multivitamin syrup or multivitamin syrup alone divided in three divided daily doses for a period of two weeks. Children were followed up weekly at home to assess subsequent growth and morbidity for a period of eight weeks. MAIN OUTCOME MEASURES: Gain in length and body weight and reduction in diarrhoea and respiratory tract infection. RESULTS: During the follow-up, zinc supplemented children showed significantly greater cumulative length gain (18.9 mm vs 14.5 mm, P <0.03) and comparable body weight gain than the children of the control group. Subsequent length gain was not correlated with initial height in the zinc-supplemented group (r=-0.13), P = 0.5), but was significantly correlated in the control group (r = -0.6, P < 0.0007). Zinc-supplemented and stunted children (< or = 90% length for age n = 18) experienced significantly fewer episodes of diarrhoea (0.07 vs 0.6, P < 0.05) and respiratory illness (1.0 vs 2.4, P < 0.01) compared to the control group. The underweight children (< or = 71% weight/age n = 38) receiving zinc-supplementation also had fewer episodes of diarrhoea (0.4 vs 1.0, P<0.04) and shorter duration of diarrhoeal episodes (1.0 vs 3.0d, P<0.04) compared to their counterparts in the control group. CONCLUSION: These results suggest that a short course of zinc supplementation to malnourished children during acute diarrhoea reduces growth-faltering and diarrhoeal and respiratory morbidity during subsequent two months. PMID- 10452408 TI - Effects of butter oil blends with increased concentrations of stearic, oleic and linolenic acid on blood lipids in young adults. AB - OBJECTIVE: The aim of this present project was to evaluate a more satisfactory effect on plasma lipoprotein profile of spreads based on dairy fat. DESIGN: This study was designed as a randomised cross-over experiment with a three-week treatment separated by a three-week wash-out period. Sixty five grams of the fat content of the habitual diets was replaced by either butter/grapeseed oil (90:10) (BG); butter oil and low erucic rapeseed oil (65:35) (BR) or butter blended in a 1:1 ratio with a interesterified mixture of rapeseed oil and fully hydrogenated rapeseed oil (70: 30) (BS). SUBJECTS: Thirteen healthy free-living young men (age 21-26 y) fulfilled the study. INTERVENTIONS: At the beginning and end of each diet period two venous blood samples were collected. Triacylglycerol and cholesterol concentrations in total plasma and VLDL, LDL, IDL and HDL fractions were measured, as were apo A-1 and apo B concentrations. Fatty acid composition of plasma phospholipids, plasma cholesterol ester and platelets was also determined. RESULTS: Significantly (P < 0.05) lower total and LDL-cholesterol concentrations were observed after the BR and BS period, compared to BG. The effect of BR and BS did not differ. BG and BR resulted in equal concentrations of HDL-C, but significantly higher than BS. Consequently, a significantly lower LDL C/HDL-C ratio was seen after the BR treatment compared to BG and BS. Apo A-1 concentrations were not significantly different, but Apo B was significantly increased after BG. CONCLUSIONS: Partially replacing milk fat with rapeseed oil seems to yield a more healthy spread. Stearic acid had a HDL-C lowering effect compared to milk fat, but did not affect LDL-C significantly. The addition of stearic acid did not improve the plasma lipoprotein profile for young men with low cholesterol levels. PMID- 10452409 TI - Sex ratio of total energy intake in adults: an analysis of dietary surveys. AB - OBJECTIVE: The ratio of the total energy intake (TEI) reported by men and women in the same dietary survey varies considerably among dietary surveys. The purpose of our study was to investigate the potential value of the sex ratio of TEI as a measure of misreporting dietary intake by comparing it with the sex ratio of biomarkers such as the 24 h urinary excretion of sodium, potassium and total nitrogen. METHODS: The sex ratio (m/f) of TEI in adults was calculated from 81 dietary surveys performed in 28 countries. The surveys were conducted in healthy and free-living populations, using the same methodology in both sexes. RESULTS: A mean sex ratio of 1.35 (s.d., 0.13) and 1.33 (s.d., 0.10) was obtained at the individual survey level and at the country level, respectively. The sex ratio of 1.40 in the younger age class (< or = 60/64 y) was significantly higher than the sex ratio of 1.27 in the older age class (> 60/64y) (P < 0.0001). The dietary assessment methodology also influenced the sex ratio (P = 0.03). Compared with the INTERSALT study, the sex ratio of TEI was higher than the sex ratios of the 24 h urinary sodium (1.23), potassium (1.20) and total nitrogen (1.25), biomarkers of dietary sodium, potassium and protein intake. Therefore the dietary survey data indicate a mean TEI of 10 476 kJ/d for men and 7784 kJ/d for women, which gives a mean sex difference of 2692 kJ/d. However if the biomarker sex ratio of 1.23, mean sex ratio of the 24 h urinary sodium, potassium and total nitrogen, is correct and the TEI of men is correctly assessed at 10476kJ/d, the TEI of women should be 8517kJ/d, a difference of only 1959 kJ/d. CONCLUSIONS: Calculated from dietary surveys, the mean sex ratio of TEI is 1.35. This sex ratio decreases with age and depends on the dietary assessment methods used. In many dietary surveys, the sex ratio of TEI is likely overestimated. PMID- 10452411 TI - Modification of the nutrition questionnaire for elderly to increase its ability to detect elderly people with inadequate intake of energy, calcium, vitamin C and vitamin D. AB - OBJECTIVE: To validate the Nutrition Questionnaire for Elderly (NQE) aimed at health care personnel -and develop and validate a modified version of the questionnaire (MNQE) to improve its sensitivity in detecting elderly people with inadequate intake of specific nutrients and energy. DESIGN: The NQE was used to rank the nutrient intake of participants as 'acceptable' (OK-->) or 'at risk' (OBS-->) with respect to energy, calcium, vitamin C and vitamin D--nutrients with a high risk of deficiency, especially among the homebound or institutionalised elderly. The sensitivity, specificity and predictive capacity were calculated by means of a 4 d estimated record (study 1). Based on the results minor adjustments of the NQE was made, and the already obtained information was used to assess the theoretical validity of the MNQE (study 2). Finally the MNQE was validated in a new population (study 3). SETTING AND SUBJECTS: Ninety-five old people ( > 65 y) living in nursing-homes (Bagsvaerd or Havdrup), or at home in the same home-care district (Valby) or in the same preventive home-visit district (Rodovre) were recruited. Excluded were those who declined to participate or who were in a terminal condition (n = 3). RESULTS: The sensitivity of the NQE regarding the intakes of energy, calcium, vitamin C and vitamin D was quite low (respectively 0.46, 0.08, 0.05 and 0.38) (study 1). However minor adjustments of the NQE increased the sensitivity both in theory (energy: 0.64, calcium: 0.95, vitamin C: 1.00 and vitamin D: 0.84) (study 2) and in practice (energy: 1.00, calcium: 0.40, vitamin C: 0.86 and vitamin D: 0.85) (study 3). Modification of the NQE did not reduce the high specificity and predictive capacity of the original version. CONCLUSIONS: Modifications of the NQE resulted in a questionnaire with a high sensitivity, specificity and predictive capacity for detecting inadequate intake of energy, calcium, vitamin C and vitamin D in elderly people. PMID- 10452410 TI - Iron deficiency in older people: interactions between food and nutrient intakes with biochemical measures of iron; further analysis of the National Diet and Nutrition Survey of people aged 65 years and over. AB - OBJECTIVE: To quantitatively investigate relationships between food consumption, nutrient intake and biochemical markers of iron status in a population of older people. DESIGN: National Diet and Nutrition Survey: cross-sectional survey of a nationally representative population of men and women aged 65 y and over. SETTING: Mainland Great Britain SUBJECTS: 1268 participants (651 men and 617 women) who provided both dietary intake and blood biochemistry. 986 were living in private households and 282 were living in residential or nursing homes. RESULTS: Intakes of alcohol, vitamin C, protein, haem and non-haem iron and fibre were positively associated with iron status. Consumption of meat, poultry and fish were positively associated with six measures of iron status, and vegetables and potatoes with four measures. Calcium, dairy foods and tea generally had a negative association with most measures of iron status. CONCLUSIONS: A varied diet containing meat, poultry and fish, vegetables and fruit, with a moderate intake of alcohol will make a positive contribution to the iron status of elderly people. PMID- 10452412 TI - The association of energy intake bias with psychological scores of women. AB - OBJECTIVE: Assess the association between reporting bias of dietary energy intake and the behavioral and psychological profiles in women. DESIGN: At baseline a series of questionnaires were administered to 37 women, (the Marlowe-Crowne Social Desirability Scale, Weinberger Adjustment Inventory (WAI), the Eating Disorder Inventory (EDI), the Restraint Scale and Sorensen-Stunkard's silhouettes). Subjects received training on how to record dietary records. Subjects recorded three days of dietary records to measure energy intake (EI) during a study to determine total energy expenditure (TEE) using doubly labeled water. Reporting accuracy (RA = EI/TEE x 100) was determined for each subject. Statistical analysis of the data used a mixed effects model accounting for within subject variability to determine if the psychological scores were associated with reporting accuracy. SETTING AND SUBJECT: Women were recruited with local advertisements in Tucson, Arizona. The women had a mean ( +/- 1 s.d.) age of 43.6 +/- 9.3 yrs, body mass index (BMI) of 28.7 +/- 8.5 kg/m2 and total body fat (%TBF) of 31.9 +/- 7.3%. RESULTS: Age and %TBF were significantly and inversely associated with RA. Furthermore, Social Desirability was negatively associated with RA. Body dissatisfaction and associating a smaller body size than one's own as being more healthy were also associated with a lower RA. CONCLUSIONS: These results suggest that Social Desirability and self image of body shape are associated with RA. Modifications in subject training may reduce the effect of these factors on RA. PMID- 10452413 TI - Nutritional status of adult inpatients in Bujumbura, Burundi (impact of HIV infection). AB - OBJECTIVE: To determine the prevalence and analyse the characteristics and causes of malnutrition among inpatients in an HIV highly endemic area. DESIGN: Prospective cross-sectional study. SETTING: Department of Internal Medicine, University Hospital, Bujumbura, Burundi. SUBJECTS: 226 adult inpatients. MAIN OUTCOMES MEASURES: Evaluation of nutritional status using anthropometry. Degree of malnutrition defined by the percentage of body weight loss (BWL), calculated by reference to the usual body weight. RESULTS: Among the 226 patients recruited (mean age: 34.4 +/- 11.9 y, M:F sex ratio: 1.72), 102 (45.1%) were HIV seropositive. 62 (60.8%) of these HIV seropositive were AIDS cases. The AIDS defining criterion was 'wasting syndrome' for 25 (40.3%) and opportunistic infection (OI) for 37 (59.7%) including 34 cases of tuberculosis (TB). The nutritional status of 119/226 patients (52.7%) was normal (BWL < or = 10%). Moderate malnutrition (10% < or = BWL < or = 20%) was observed in 47 (20.8%) and severe malnutrition (BWL > 20%) was observed in 60 (26.5%). HIV seroprevalence and, among HIV seropositive subjects, the percentage of AIDS cases increased according to decreasing level of nutrition (Chi2 for trends: P < 0.001 in both instances). The fat free mass mass of malnourished subjects was lower and the fat body mass was higher among HIV seropositive subjects than HIV seronegative subjects. Among HIV seropositive subjects, malnutrition was associated with TB (P < 0.001) and dysphagia (P < 0.05). Among HIV seronegative subjects, malnutrition was associated with decreased food availability (P < 0.003) and TB (P < 0.05). One week after admission, the mortality rate was higher among HIV seropositive subjects (10.8%) than seronegative subjects (2.4%, P=0.009). Other factors associated with death were decreased fat free mass (P < 0.01) and tricipital skinfold thickness (P < 0.04). CONCLUSIONS: The prevalence of malnutrition is high among the inpatients investigated. Main factors are HIV infection and TB. Strategies adapted to the African context should be developed to prevent, detect and treat malnutrition and associated factors, particularly among HIV seropositive subjects. PMID- 10452414 TI - Two good reasons: women's and men's perspectives on dual contraceptive use. AB - In the US, continued high rates of unintended pregnancy, combined with increases in heterosexual transmission of HIV to women, have sharply magnified concern about the factors leading to or barring the use of contraceptive methods to protect concurrently against both risks. This paper reports on results of focus group research among African-American women participating in a longitudinal study and African-American men who are either partners of the women or are of similar socio-economic status as their partners. We found a high level of agreement between men and women on the issues and problems that both sexes face. People felt that regardless of a woman's use of other contraceptive methods, a condom should always be used for protection. This belief, however, differed markedly from actual practice. Although we attempted to discern the relative salience of concern about pregnancy versus STIs, we conclude that people may not separate these two concerns in their resolve to use two methods. Furthermore, they recognized the need for dual protection, but expected conflict with their partners from using condoms as a second method because of high levels of distrust regarding sexual fidelity. Thus people are caught in a bind: distrust further increases the sense of a need for dual methods, but using condoms exacerbates the problems people have with achieving trust in relationships. PMID- 10452415 TI - Policy and politics of smoking control in Japan. AB - The tobacco industry took root well before the hazards of its products were proven scientifically. As elsewhere, smoking control policy has not proceeded automatically nor smoothly in Japan. Examination of the past political process discloses that the failure to enact effective smoking control is attributable to several factors, including the political environment, administrative inadequacy and an inactive medical community. Especially remarkable in this failure have been: the political leverage of the tobacco industry; a lack of clear leadership by the health ministry; the successive rejection of law suits in the courts; and the relatively weak health advocacy groups in Japan. The Diet had been effectively immobilized by pro-tobacco interests and without support from the government, administrative agencies have remained inactive. Since the late 1970s, a series of smoking control measures has been introduced by government agencies, propelled by the social movement. However limited, the introduction of smoking control measures was accomplished with, or at least facilitated by the following: the advocacy of local and international groups and organizations; the continuous visibility of the issue in the media; and changing public attitudes. Involvement of the Diet and the Ministry of Health and Welfare (MHW) was essential in translating commitments into government action. Nevertheless, the resulting measures were not comprehensively legislated. Moreover, they were not subject to continuous evaluation. Consequently, they have been mostly ineffective in decreasing the prevalence of smoking among the populace. Clearly, leadership by the health ministry coupled with political support are the key to advancing effective smoking control. PMID- 10452416 TI - Knowledge, beliefs and practices relevant for malaria control in an endemic urban area of the Colombian Pacific. AB - Research projects for malaria control must involve communities to elicit strategies to be successful and programs to be sustainable. Therefore, knowledge, beliefs and practices of the population concerned must be taken into account in the design of interventions against malaria transmission. We conducted a KAP study in Buenaventura, a port on the Pacific Coast of Colombia where transmission was on the increase at the beginning of this decade. The purpose of the study was to help in the design and implementation of a primary health care approach for malaria control. Both qualitative and quantitative research methods were used. The focus group technique was applied in five urban and peri-urban communities and a cross sectional survey was conducted on a random sample of 1380 subjects with a structured interview on knowledge, practices and also on demographic and epidemiological aspects. The information obtained by the two methods was comparable on knowledge of symptoms, causes and ways of malaria transmission, and prevention practices like the use of bednets or provision of health services. We discuss the relevance of obtaining this information and analyze the elements that must be taken into account for communities to become more involved in malaria control strategies. PMID- 10452417 TI - Health behaviour and the school environment in New South Wales, Australia. AB - The relationship between the school environment and health has infrequently been examined. This study sought to examine the association between school students' perceptions of their school environment, teachers' and peers' support and their health behaviours. A cross sectional descriptive survey by supervised self administration was conducted in 1996 based on the international WHO collaborative survey of school children's health and lifestyle (the HBSC Study) and extended in an Australian setting. Randomly sampled primary and secondary schools from Catholic, Independent and Government education sectors throughout New South Wales (NSW), Australia, were invited to participate. The final sample included 3918 school students attending Year 6 (primary school), Year 8 and Year 10 (high school) from 115 schools. The main outcome measures were self-reported health status and 7 health behaviours (tobacco use, alcohol use, physical activity, dental hygiene, nutritional intake, seat belt and bicycle helmet use). Independent variables included student perceptions of the school environment, perceptions of teachers' and peers' support. Girls, Year 6 students and students who have less than $19 a week to spend were significantly more likely to have positive perceptions towards their school environment, teacher(s) and peers. Students who had positive perceptions regarding their school environment and perceived their teachers as supportive were significantly more likely to engage in health promoting behaviours adjusting for age, sex and average weekly pocket money. A supportive peer environment was not associated with positive health behaviour. Health promotion practitioners need to consider the impact of the school environment on health behaviours of school students. In particular, practitioners should consider intervention models that improve the school environment as a key strategy within a health promoting school. PMID- 10452418 TI - Age differences in the effects of network composition on psychological distress. AB - The main goal of this research is to better understand age differences in the effects of social networks on mental health. Using a social network approach to revise the convoy of social support model (Kahn and Antonucci, 1980), we examine specifically how two aspects of social support networks (kin composition and convoy dimensions) influence psychological distress for older and younger samples (18-59 and 60+). We hypothesize that kin composition will influence distress in general but especially for the younger sample, while two competing hypotheses for the convoy of social support model are tested. Using data from a three-wave panel health study, structural equation models (LISREL 8.20) indicate that a greater proportion of kin in the perceived support network and the presence of family members in the inner circle of the convoy significantly reduce distress, primarily for the younger sample. Implications for the convoy model are discussed. PMID- 10452419 TI - The quest of infertile women in squatter settlements of Karachi, Pakistan: a qualitative study. AB - OBJECTIVES: There is a dearth of knowledge regarding Pakistani women's perceptions and treatment seeking behavior for infertility. We conducted this study to explore the contextual factors that influence the health-seeking behavior of infertile women in the lower socio-economic group. METHODS: In-depth interviews were conducted with 17 women, identified from clinics and community using pre-tested interview guidelines covering issues as the chronological sequence of the number and types of the health care providers consulted for infertility, household dynamics regarding decision making, perceived causes and social and psychological effects of infertility on the woman and her family. RESULTS: 14 of the 17 women interviewed presented with primary infertility and three with secondary infertility. Reasons reported for seeking treatment for primary infertility were: someone to carry on the family name, feeling alone and akhrat par maan bap bun kar uthna (getting up as a parent on judgment day). Two of the three women suffering from secondary infertility were seeking treatment because they did not have any male offspring. Of the 14 women, 11 initially sought treatment within the first two years of marriage; one woman reported that she was "coerced" by her mother-in-law to seek treatment following the first week of marriage. On average, a woman went to three health care providers (general practitioners, gynecologists and Traditional Birth Attendants) in her quest for assistance. The effects that infertility had on these women ranged from social pressure, coercion by in-laws and/or social isolation. Only one woman reported being put off by sex because it was not giving her what she 'desired'. CONCLUSION: Women in Pakistan seek care for infertility early and go to various types of health care providers. Infertility is usually treated as a clinical disease by the health care providers without considering its social ramifications. PMID- 10452420 TI - Decision-making in the physician-patient encounter: revisiting the shared treatment decision-making model. AB - In this paper we revisit and add elements to our earlier conceptual framework on shared treatment decision-making within the context of different decision-making approaches in the medical encounter (Charles, C., Gafni, A., Whelan, T., 1997. Shared decision-making in the medical encounter: what does it mean? (or, it takes at least two to tango). Social Science & Medicine 44, 681 692.). This revised framework (1) explicitly identifies different analytic steps in the treatment decision-making process; (2) provides a dynamic view of treatment decision-making by recognizing that the approach adopted at the outset of a medical encounter may change as the interaction evolves; (3) identifies decision-making approaches which lie between the three predominant models (paternalistic, shared and informed) and (4) has practical applications for clinical practice, research and medical education. Rather than advocating a particular approach, we emphasize the importance of flexibility in the way that physicians structure the decision making process so that individual differences in patient preferences can be respected. PMID- 10452421 TI - Evaluating health service equity at a primary care clinic in Chilimarca, Bolivia. AB - Policy makers and health planners generally support the concept of equitable health care. A focus on who can use a health service, or its potential access, will not necessarily lead to equitable care if people are not willing to avail themselves of the health services offered. Because equity is difficult to operationalize, outcome-based indicators such as the actual utilization of services are advocated as a means to measure equal access. This paper evaluates the utility of linking the concept of equity with a temporal and spatial analysis of clinic users at a micro scale, supplemented by a community survey. Various spatial scales were employed in the analysis. Utilization of the primary care clinic in Chilimarca, Bolivia varied considerably during the first 25 months of operation. Spatially, utilization shifted away from the targeted service area. Within the targeted service area, usage was concentrated in a few blocks of the community and generally diminished with increasing distance from the clinic. The survey further revealed place of origin, length of residence, and language spoken at home as variables differentiating users from non-users. Failure to include the spatial dimension of utilization would lead to different conclusions if only aggregate data were employed. Spatial analysis of output measures is imperfect and does not necessarily deal with all of the access issues related to acceptability. They do, however, begin to isolate areas of a defined geographic area where further investigation would assist in ascertaining, and subsequently addressing, potential problems related to equal access. PMID- 10452422 TI - National health care 'by-passing' in Bangladesh: a comparative study. AB - Using data from a survey of urban residents of Dhaka, Bangladesh, the phenomenon of by-passing national health care resources in favor of foreign ones has been analyzed. The results clearly suggest that the incidence of national health care by-passing is relatively widespread, particularly among the wealthy residents of Dhaka. The findings of this study show that the health care system in Bangladesh is costly, some specialized care is not available, and the quality of services obtained in foreign countries is perceived as better than what is available nationally. While complete prevention of national health care by-passing is not possible, improvement in health care services available within Bangladesh will help in decreasing the incidence of by-passing. PMID- 10452423 TI - Revisiting medicalisation and 'natural' death. AB - The contemporary conceptualisation of natural death in social science and health care literature may be seen as elision of potentially paradoxical ideas in which the process of dying, as opposed to the moment of death, is a key determinant of the manner in which death is regarded. In the predominant rhetoric, medical technological intervention during dying is emblematic of inhumane and unnatural death. Highly technological clinical settings, where medical intervention in the process of dying is so clearly visible, are held up as extreme examples of the metamorphosis of death from 'natural' into 'unnatural' events. This paper examines the reification of 'natural' death within these writings, focusing on the taken for granted polarisation of technology and 'natural' death with which they are underpinned. The paper then turns to an assessment of the validity of this reification by examining some ethnographic case study data concerning the experiences of the close companions of three people who died, or came near to death, within intensive care: arguably an environment in which death is at its most highly medicalised. The data, which are drawn from a wider ethnography of death and dying in two general adult intensive care units, suggest that it is perceptions of the meaning of technology, rather than its simple minimisation or absence, which determine representations of death within highly technological settings. These perceptions in their turn depend crucially on the circumstances with which dying is attended. In this study the 'natural' process of death was preserved for the companions of dying people when medical technology delivered the outcomes they expected, appeared to be amenable to human manipulation and intention, was accessible to their understanding and seemed to 'fit' with the wider context of the dying person's life. The paper concludes by arguing that it is within the phenomenology of suffering associated with the critical illness or death of a close companion that some insights may be gleaned of the relationship between individual experience, the cultural representation of 'natural' death, and the attitudinal ambivalence with which medical technology is surrounded. PMID- 10452424 TI - Plain abdominal radiography in clinically suspected appendicitis: diagnostic yield, resource use, and comparison with CT. AB - This study determined the diagnostic utility and hospital resource impact of plain abdominal radiography in emergency department patients with suspected appendicitis. The authors reviewed medical records of 821 consecutive patients hospitalized for suspected appendicitis; 78% had plain abdominal radiography. Sixty-four percent had appendicitis. Radiographic findings were noted in 51% of patients with, and 47% of patients without appendicitis; no individual radiographic finding was sensitive or specific. Specific conditions were suggested in 10% of impressions; these failed to correlate with final clinical diagnoses 57% of the time. Hospital cost per abdominal radiograph was $67; cost per specific, correct radiographic diagnosis was $1,593. This is compared with $270 per appendiceal computed tomography scan (based on recent literature data). The authors conclude that plain abdominal radiographs in patients with suspected appendicitis are neither sensitive nor specific, are frequently misleading, are costly per specific and correct diagnosis, and should not be routinely obtained on patients with suspected appendicitis. PMID- 10452425 TI - Prognostic factors in acute cardiogenic pulmonary edema. AB - The purpose of this study was to determine the clinical and biological findings at admission in the Department of Emergency Medicine associated with a poor prognosis, and to evaluate early response to treatment as a prognostic factor. It was a prospective cohort study with a 5-month follow-up. One hundred eighty-six patients admitted for acute cardiogenic pulmonary edema were included. Features were analyzed at the admission and on response to initial treatment. The main outcome measure was survival at 2 end-points: hospital discharge, and 5 months of follow-up. Multivariate analysis showed that in-hospital mortality was associated with marbleization (mottling) odd-ratio (OR) = 9.0), low diuresis (OR = 4.0), high breath rate 6 hours after admission (OR = 4.0), and chronic digoxin use (OR = 3.39). Five-month mortality was associated with a bedridden state (OR = 9.0), marbleization (mottling) (OR = 5.5), myocardial infarction (OR = 3), and poor early response to initial treatment (OR = 3.2). In addition to well-known factors, the response to initial treatment evaluated 6 hours after admission was a major determinant of outcome. PMID- 10452426 TI - Prehospital stability of diazepam and lorazepam. AB - Injectable benzodiazepines are commonly stocked on ambulances for use by paramedics. We evaluated the stability of lorazepam and diazepam as a function of storage temperature. Diazepam (5 mg/mL) and lorazepam (2 mg/mL) injectable solutions were stored for up to 210 days in clear glass syringes at three conditions: 4 degrees C to 10 degrees C (refrigerated); 15 degrees C to 30 degrees C (on-ambulance ambient temperature); and 37 degrees C (oven-heated). High-performance liquid chromatography (HPLC) analyses of syringe contents were performed at 30-day intervals. After 210 days, the reduction in diazepam concentration was 7% refrigerated, 15% at ambient temperature, and 25% at 37 degrees C. The reduction in lorazepam concentration was 0% refrigerated, 10% at ambient temperature, and 75% at 37 degrees C. Whereas diazepam retained 90% of its original concentration for 30 days of on-ambulance storage, lorazepam retained 90% of its original concentration for 150 days. The decrease in lorazepam concentration correlated with an increase in the maximum ambient temperature in San Francisco. These results suggest that diazepam and lorazepam can be stored on ambulances. When ambient storage temperatures are 30 degrees C or less, ambulances carrying lorazepam and diazepam should be restocked every 30 to 60 days. When drug storage temperatures exceed 30 degrees C, more frequent stocking or refrigeration is required. PMID- 10452427 TI - Localizing ingested coins with a metal detector. AB - This study was conducted to determine the utility of metal detection in coin localization by inexperienced operators, and determine the rate of spontaneous passage of asymptomatic esophageal coins. All children who presented to the emergency department of an urban children's hospital with a suspected coin ingestion were eligible. Coin location was predicted from metal detector results, while radiographs confirmed location. Asymptomatic patients with esophageal coins were observed for spontaneous passage. Ninety-one children (ages 9 months to 17 years) were prospectively enrolled. The metal detector had a sensitivity of 98% (53/54) in coin detection and 98% (81/83) in determining coin location as esophageal. Symptoms were poor predictors of coin location. Six of eight asymptomatic patients with esophageal coins spontaneously passed their coins. These results show that metal detection is a good screening test for coin presence and to determine coin location as esophageal. Spontaneous passage of asymptomatic esophageal coins warrants further study. PMID- 10452428 TI - Abdominal distention and shock in an infant. AB - Acute abdominal distention in the pediatric patient may be attributable to extraperitoneal fluid, masses, organomegaly, air, an ileus, a functional or mechanical bowel obstruction, or injury and blood secondary to trauma. An infant who presents to the emergency department with acute abdominal distention and shock is a true emergency for which the differential diagnosis is extensive. An unusual case of abdominal distention, ascites, hematochezia, and shock in an infant, subsequently found to have spontaneous perforation of the common bile duct is reported. This uncommon cause of abdominal distention and shock in an infant is many times left out of the differential diagnosis of an acute abdomen. The presentation may be as an uncommon acute form or a classis subacute type. This patient had hematochezia, which had not been previously reported in association with this entity. Failure to recognize and treat an acute abdomen can result in high mortality. PMID- 10452429 TI - Painless intussusception and altered mental status. AB - A 7-month-old child presented to the emergency department (ED) with 2 hours of painless, nonprojectile emesis and a normal mental status. Over a 3-hour period in the ED, the child remained pain-free, but developed hematemesis, hematochezia, and lethargy, progressing to unresponsiveness. The patient was evaluated for toxic ingestion, intracranial bleed, sepsis/meningitis, and intraabdominal pathology. The diagnosis was made by an abdominal ultrasound, which demonstrated an ileal-cecal intussusception that ultimately required surgical reduction. This case illustrates an insidious and poorly understood presentation of a common childhood affliction, as well as the utility of abdominal ultrasound in evaluating a hemodynamically stable patient with intussusception. PMID- 10452430 TI - Thyroid gland hematoma after blunt cervical trauma. AB - Thyroid hematoma is a rare cause of airway obstruction in victims of blunt trauma. The case of a 34-year-old woman who developed orthopnea after a low energy motor vehicle accident is described. Presenting greater than 24 hours after her accident, the patient noted dysphagia, tracheal deviation, and postural dyspnea. The diagnosis of thyroid gland hematoma was made with a combination of fiberoptic laryngoscopy, cervical computed tomography, and great vessel and carotid angiography. Invasive airway management was not required. The patient underwent a total thyroidectomy and recovered without complications. PMID- 10452431 TI - Tension pneumoperitoneum caused by blunt trauma. AB - Tension pneumoperitoneum (TPP), the accumulation of free intraabdominal air under pressure, is a rare event. TPP usually occurs from bowel surgery or bowel perforations. Less commonly, TPP occurs in the presence of pneumothoraces or during positive pressure ventilation. Trauma has rarely been a reported cause of TPP. The cases of 2 patients with TPP after blunt trauma are reported. The pathophysiology and management of TPP are discussed. PMID- 10452432 TI - Testing for HIV: current practices in the academic ED. AB - The objective of this study was to determine common practices for testing for Human Immunodeficiency Virus (HIV), particularly in patients with other sexually transmitted diseases (STD) in emergency departments (ED) with residency training in Emergency Medicine. Via mail, 112 directors of academic emergency medicine programs in the United States were surveyed. Surveys from 95 academic institutions were completed, returned, and included in the analysis. Three EDs (3%) routinely tested for HIV in patients with suspected STD. HIV testing was performed in the ED in 54% of responding institutions under special circumstances such as employee testing after occupational exposures (54%), cases of rape (46%), and suspicion of HIV infection by clinical manifestations other than suspected STD (36%). Based on the results it was determined that academic EDs do not routinely test for HIV in patients suspected of having a STD and have variable testing practices and policies regarding other possible HIV exposures. PMID- 10452433 TI - Boxer's fracture: an indicator of intentional and recurrent injury. AB - The objective was to describe the population of patients presenting with a boxer's fracture (BF), to determine how often BF is an intentional injury, to determine if it is a predictor of recurrent injury, and to compare the rates of intentional injury and injury recidivism between patients with BF and those with other injuries. An emergency department (ED)-based injury surveillance system (EDBISS) in a university-affiliated ED (census of 35,000) provided data on injured patients presenting between January 1, 1995 and December 31, 1996. Chart review was performed. Descriptive analyses were performed. The results showed 22,728 of 69,438 (33%) ED visits during a 2-year period were attributable to injuries. Sixty-two patients presented with BF. Mean age of BF patients was 22.1 versus 28.6 for all injured (P < .005). Ninety-two percent (57/62) of BF patients were men compared with 58% of all injured (P < .0001). Thirty-eight of 62 (61%) BF injuries were sustained after intentional punches of an object/person. Seventeen of 62 (27%) BF patients were injury recidivists. Previous studies in this same ED population showed that 6% had intentional injuries and 12% were injury recidivists. BF is usually an intentional injury and these patients are at increased risk for recurrent injury. Clinicians should focus prevention efforts on this high-risk population. PMID- 10452434 TI - Painless aortic dissection presenting as hoarseness of voice: cardiovocal syndrome: Ortner's syndrome. AB - Most of the neurological manifestations of the aortic dissection are due to neuronal ischemia secondary to either extension of the dissection process into a branch artery, or compression of an artery by the false lumen of the dissecting aortic hematoma. However, the enlarging false lumen may directly compress on an adjacent nerve, causing neuronal injury resulting in neurological symptoms. This may particularly take place when a distal intimal tear does not decompress the false lumen, resulting in formation of an expanding blind pouch. About 10% of aortic dissections are painless and may present with symptoms secondary to the complications of the dissection. Although cardiovocal syndrome, or Ortner's syndrome (hoarseness of voice due to involvement of recurrent laryngeal nerve in cardiovascular diseases) has been described with aortic dissection, it has not been reported as an initial presenting feature of this disorder. This report describes the first case of painless aortic dissection presenting with hoarseness of voice, the cardiovocal syndrome. The hoarseness remained the only symptom throughout the entire course of the disease. The aortic dissection was not suspected initially. During surgical exploration, the recurrent laryngeal nerve was found compressed by the false lumen at the level of aortic arch. Aortic root replacement was performed successfully, resulting in complete resolution of the hoarseness. The neurological manifestations of aortic dissection, and the cardiovocal syndrome, are discussed. PMID- 10452435 TI - pH-dependent cocaine-induced cardiotoxicity. AB - Severe cocaine toxicity causes acidemia and cardiac dysfunction. These manifestations are described in 4 patients who presented with seizures, psychomotor agitation, and cardiopulmonary arrest. Their initial laboratory values demonstrated acidemia and electrocardiographic findings that included a prolonged QRS complex and QTc duration and a rightward T40 ms axis deviation. Treatment of the patients with hyperventilation, sedation, active cooling, and sodium bicarbonate infusion led to the normalization of their blood pHs and reversal of their cardiac conduction disorders. Acidemia can contribute to cocaine cardiac disorders by promoting conduction delays, dysrhythmias, and depressed myocardial contractility. Good supportive care corrects the blood pH and cardiac conduction disorders and remains the major focus in the management of patients with cocaine toxicity. PMID- 10452436 TI - Intrathoracic stomach presenting as acute tension gastrothorax. AB - Total intrathoracic stomach creating pulmonary and hemodynamic compromise is a rare life-threatening complication in patients with hiatal hernia. The presentation and clinical course of this condition are discussed. Physicians should consider this entity in patients presenting with apparent tension pneumothorax without history or other evidence of trauma or positive pressure ventilation who do not respond to standard interventions. PMID- 10452437 TI - Aortic dissection. AB - The presentation of aortic dissection in the emergency department may be more subtle than the classic description of a shocked patient with "ripping" chest pain. The epidemiology, variation in presentation, investigation, and management of aortic dissection are reviewed. PMID- 10452438 TI - Wide QRS complex tachycardia: ECG differential diagnosis. AB - Wide QRS complex tachycardias (WCT) present significant diagnostic and therapeutic challenges to the emergency physician. WCT may represent a supraventricular tachycardia with aberrant ventricular conduction; alternatively, such a rhythm presentation may be caused by ventricular tachycardia. Other clinical syndromes may also demonstrate WCT, such as tricyclic antidepressant toxicity and hyperkalemia. Patient age and history may assist in rhythm diagnosis, especially when coupled with electrocardiographic (ECG) evidence. Numerous ECG features have been suggested as potential clues to origin of the WCT, including ventricular rate, frontal axis, QRS complex width, and QRS morphology, as well as the presence of other characteristics such as atrioventricular dissociation and fusion/capture beats. Differentiation between ventricular tachycardia and supraventricular tachycardia with aberrant conduction frequently is difficult despite this clinical and electrocardiographic information, particularly in the early stages of evaluation with an unstable patient. When the rhythm diagnosis is in question, resuscitative therapy should be directed toward ventricular tachycardia. PMID- 10452439 TI - Acute flank pain: an unusual presentation of a spinal AVM. AB - The authors report the case of a 6-year-old boy with a spinal cord arteriovenous malformation (AVM) who presented with acute flank pain and a delayed onset of paraplegia. An early diagnosis of a spinal cord AVM was made difficult by the absence of neurological findings on initial evaluation. Included is a description of his clinical course, and the presentation of spinal AVMs to the emergency physician is discussed. PMID- 10452440 TI - Lacrimal canaliculitis. AB - Lacrimal canaliculitis is an infection of the lacrimal duct system. The classic features of lacrimal canaliculitis are mild to severe swelling of the canaliculus, mucopurulent discharge from the punctum, and a red, pouting punctum. Canaliculotomy with systemic or topical antibiotics is the most appropriate treatment for this disorder. PMID- 10452441 TI - ECG abnormalities in tricyclic antidepressant ingestion. AB - The tricyclic antidepressant (TCA) agents are recognized for their potentially lethal cardiovascular and neurological effects in poisoned patients. The 12-lead electrocardiogram (ECG) has emerged as a popular bedside tool in the evaluation of TCA toxicity. Although the history and physical examination play a key role in the assessment of the patient with potential TCA poisoning, the presence or absence of features of the TCA toxidrome are not sufficient to detect or exclude toxicity from this class of drugs. A variety of ECG findings occur with TCA toxicity. Aside from the sinus tachycardia due principally to anticholinergic effects, TCA-toxic changes seen on the ECG are attributable primarily to the sodium channel blockade caused by these agents. The majority of patients at significant risk for developing cardiac or neurological toxicity will have a QRS complex greater than 0.10 seconds or a rightward shift of the terminal 40 ms of the frontal plane QRS complex vector. The majority of these patients will also display these changes early in their emergency department stay. However, the appearance of these findings, either alone or in combination, does not mean the patient will develop significant cardiac or neurological toxicity. The ECG can neither unequivocally rule in nor rule out impending toxicity; recognizing these limitations, the emergency physician can use this bedside tool in combination with other clinical data during the assessment of the poisoned patient. PMID- 10452442 TI - The nondiagnostic ECG in the chest pain patient: normal and nonspecific initial ECG presentations of acute MI. AB - The 12-lead electrocardiogram (ECG) is a powerful clinical tool used in the evaluation of chest pain patients, assisting in the selection of the proper therapy. Unfortunately, the ECG is diagnostic of acute myocardial infarction (AMI) in only one-half of such patients at initial hospital evaluation. In the remaining group of patients with the nondiagnostic 12-lead electrocardiogram, the ECG may be entirely normal, show nonspecific sinus tachycardia (ST) segment-T wave abnormalities, or obvious ischemic changes. In adult chest pain patients treated in the emergency department (ED), 1% to 4% of such patients with an absolutely normal ECG had a final hospital diagnosis of AMI; furthermore, patients with nonspecific electrocardiographic abnormalities experienced AMI in 4% of cases. These findings reinforce the teaching point that the history is the most important tool used in the evaluation of chest pain patients. Furthermore, overreliance on a normal or nonspecifically abnormal ECG in a patient with a classic description of anginal chest pain is dangerous. PMID- 10452443 TI - Droperidol for acute migraine headache. AB - The use of intramuscular droperidol to treat acute migraine headache has not been previously reported in the emergency medicine literature. It is a promising therapy for migraine. The authors performed a pilot review of all patients receiving droperidol for migraine in our emergency department (ED) to evaluate its efficacy. We used a retrospective case series, in a suburban ED with an annual patient census of 48,000. All patients with a discharge diagnosis of migraine headache who were treated with i.m. droperidol during a consecutive 5 month period in our ED were identified. All patients received droperidol 2.5 mg intramuscular. As per ED protocol, their clinical progress was closely followed and documented at 30 minutes after drug administration (t30). Demographic and clinical variables were recorded on a standardized, closed-question, data collection instrument. The primary outcome measurement was relief of symptoms at t30 to the point that the patient felt well enough to go home without further ED intervention (symptomatic relief). Thirty-seven patients were treated (84% female), with an ED diagnosis of acute migraine with droperidol during the study period. The mean age was 36 +/- 12 years. Analgesics had been used within 24 hours before ED presentation by 62% of patients. At t30, 30 (81%) patients had symptomatic relief, 2 (5%) felt partial relief but required rescue medication, and 5 (14%) had no relief of symptoms. Drowsiness (14%) and mild akathisia (8%) were the only adverse reactions observed following drug administration. Droperidol 2.5 mg intramuscular may be a safe and effective therapy for the ED management of acute migraine headache. Randomized, controlled trials are warranted to further validate the findings of this preliminary study. PMID- 10452444 TI - Current concepts in the treatment of anterior shoulder dislocations. AB - Anterior shoulder dislocations are commonly seen in emergency departments. With the recent proliferation of shoulder arthroscopy, the pathoanatomy has been better delineated. Arthroscopic series have confirmed the very high percentage of Bankart lesions (avulsions of the inferior glenohumeral ligament-labral complex), especially in younger patients. A high rate of recurrent dislocation in young patients has been noted in the literature with standard conservative treatment, consisting of immobilization with or without rehabilitation. This high recurrence rate is thought to be due to the Bankart lesion. Recently, investigations with the use of arthroscopic Bankart repairs have shown high success rates in preventing recurrences, with low surgical morbidity. A young, first-time, traumatic anterior dislocation patient should be referred for consideration of possible early arthroscopic Bankart repair, rather than automatically being treated conservatively with immobilization. PMID- 10452445 TI - Emergency department-based telemedicine. AB - Although many studies have been published concerning clinical telemedicine, little information is available about emergency department (ED)-based telemedicine programs. An ED-based telemedicine program was initiated in April 1996 involving the National Cheng Kung University Hospital (NCKUH) and the Provincial Peng-Hu Hospital (PPHH) under a pilot project supported by the Department of Health. This is the first telemedicine program for remote offshore island service in Taiwan. The program is synchronous in nature to the practice of telemedicine. The role of the emergency physician includes giving initial suggestions, arranging consultations, coordination, and the organization of other medical tasks, such as accompanying some of the transfers. During the 12-month period, this system was used in 275 consultations, including 24 specialty and/or subspecialty department/sections, and more than 100 members of the medical staff have participated in this project since. In a survey, 89.4% of physicians in the PPHH and 82.2% of the physicians in NCKUH rated the system as very comfortable to work with and satisfactory. According to these observations, an ED-based telemedicine program is a feasible method for carrying out remote consultations. Successful development of the partnership and program of telemedicine is based on the active participation and coordination of the medical personnel and technicians between the cooperating hospitals. PMID- 10452446 TI - Emergency medicine in Eritrea: rebuilding after a 30-year war of independence. AB - Eritrea became a member of the United Nations in 1993, after a devastating 30 year war of independence with Ethiopia. Although Eritrea suffers from many of the problems beset by the developing world, the people are determined to rebuild their country. Emergency medical care and emergency services are in their infancy, yet the signs of progress are already visible in the capital city of Asmara. This report describes the current state of the prehospital system and emergency medical care in Eritrea. PMID- 10452447 TI - Trauma: an annotated bibliography of the recent literature, 1998. PMID- 10452448 TI - Bronchial injury and pulmonary edema caused by hydrogen sulfide poisoning. PMID- 10452449 TI - Acute abdomen due to spontaneous torsion of an accessory spleen. PMID- 10452450 TI - Seizure and hyponatraemia after overdose of trazadone. PMID- 10452451 TI - Management of acute alcoholic intoxication. PMID- 10452452 TI - Medial pneumothorax: a radiographic sign that should not be overlooked on the supine view. PMID- 10452453 TI - Cheerleading as a cause of splenic rupture. PMID- 10452454 TI - Neurobiology of gender differences in reported symptoms for acute myocardial infarction. PMID- 10452455 TI - Evidence-based medicine in trauma care: whither goest thou? PMID- 10452456 TI - Dry fibrin sealant dressings reduce blood loss, resuscitation volume, and improve survival in hypothermic coagulopathic swine with grade V liver injuries. AB - OBJECTIVE: The majority of early trauma deaths are caused by uncontrolled hemorrhage, and are frequently complicated by hypothermic and dilutional coagulopathies. Any hemorrhage-control technique that achieves rapid hemostasis despite a coagulopathy should improve the outcome of these patients. We conducted this study to determine whether dry fibrin sealant dressings (DFSD) would stop bleeding from grade V liver injuries in swine that were hypothermic and coagulopathic. METHODS: Nineteen swine weighing 39.7 kg (mean and 95% confidence interval, 36.3-43.1), underwent a 60% isovolemic, hypothermic exchange transfusion with 33 degrees C 6% hetastarch to produce a dilutional and hypothermic coagulopathy. The animals then received a grade V liver injury and one of three treatments: DFSD, conventional liver packing with gauze sponges, or immunoglobulin G (IgG) placebo sealant dressing (blinded control). All animals were resuscitated with lactated Ringer's solution to their preinjury mean arterial pressure. Blood loss after treatment, mean arterial pressure, resuscitation volume, hematologic variables, and core temperature were monitored for 1 hour. RESULTS: At the time of injury, core temperature = 33.3 degrees C (95% confidence interval, 33.2-33.4), hemoglobin concentration = 4.4 g/dL (4.2 4.6), platelet count = 132 x 10(5)/microL, (93-171), prothrombin time = 21.6 seconds (19.6-23.5), activated partial thromboplastin time = 25.2 seconds (range, 22.9-27.5 seconds), and fibrinogen = 83 mg/dL (range, 76-89 mg/dL) across treatments. The posttreatment blood loss in the DFSD group was 669 mL, (range, 353-1,268 mL), which was lower (p < 0.01) than the means of 3,321 mL (range, 1,891-5,831 mL) and 4,399 mL (range, 2,321-8,332 mL) observed in the packing and IgG groups, respectively. The resuscitation volume in DFSD was 2,145 mL (range 1,310-3,514 mL), which was lower (p < 0.05) than the means of 5,222 mL (range 3,381-8,067 mL) and 5,542 mL (range 3,384-9,077 mL) in the packing and IgG groups, respectively. One-hour survival in the DFSD group was 83%, whereas survival in the packing and IgG groups were 0% (p < 0.05). CONCLUSION: In swine with a grade V liver injury complicated by a dilutional and hypothermic coagulopathy, DFSD provided simple, rapid hemorrhage control, decreased fluid requirements, and improved survival. PMID- 10452457 TI - Analysis of preventable pediatric trauma deaths and inappropriate trauma care in Montana. AB - OBJECTIVE: To determine the rates of preventable mortality and inappropriate care, as well as the nature of treatment errors associated with pediatric traumatic deaths occurring in a rural state. METHODS: Retrospective multidisciplinary consensus panel review of deaths attributed to mechanical trauma in children aged 18 years or less, occurring in Montana between October 1, 1989, and September 30, 1992. The care rendered in both preventable and nonpreventable cases was evaluated for appropriateness according to nationally accepted guidelines. Rates of pediatric preventable death and inappropriate care, as well as the nature of inappropriate care, were compared with that of the adult population. RESULTS: One hundred thirty-eight cases were reviewed. One death (less than 1%) was judged frankly preventable, 11 deaths (8%) were judged possibly preventable, giving a total preventability rate of 9% for all cases reviewed. Considering only in-hospital deaths (n = 77), the total preventability rate was 16%. The rate of inappropriate care rendered for all deaths, regardless of preventability, was 36%. The rate of inappropriate care in the prehospital phase was 16%; for in-hospital deaths, it was 47%. In the emergency department (ED), the rate was 36%, and in post-ED care, 22%. In comparison to the adult population, the rates of preventable death (9% vs. 14%) and inappropriate care in the hospital phase (64% vs. 66%) were lower. Inappropriate care for the pediatric group was more prevalent in patients less than or equal to 14 years old. The nature of inappropriate care was most frequently associated with the management of respiratory problems, including airway control and management of chest trauma. CONCLUSION: Preventable mortality from traumatic injuries in children in a rural state appears to be low, and lower than that reported for adult trauma victims in the same state. A preponderance of these preventable deaths occur in the subgroup of children less than or equal to 14 years if age. Inappropriate trauma care in children occurs frequently, particularly in the ED phase of care, and is primarily associated with the management of the airway and chest injuries. Education of ED primary care providers in basic principles of stabilization and initial treatment of the injured child 14 years old or younger may be the most effective method of reducing preventable trauma deaths in the rural setting. PMID- 10452458 TI - Blunt thoracic aortic injury: delayed or early repair? AB - OBJECTIVE: To evaluate the safety and benefit of delayed repair of blunt thoracic aortic injury (BTAI) in trauma patients with multiple injuries and to assess the financial impact of delayed repair. METHODS: A retrospective review of charts was performed on 55 patients with the diagnosis of BTAI from January 1, 1992, through December 31, 1997, at our Level I trauma center. Early repair was defined as operative repair of BTAI within 12 hours of admission. Seven patients were excluded from analysis due to death before BTAI diagnosis (two deaths were from rupture in the emergency department and five were from massive blunt trauma without rupture). The groups were compared by using a McNemar chi2 test, for which p less than or equal to 0.05 is significant. RESULTS: There were 30 patients in the early repair (ER) group repaired at 5.3+/-2.4 hours, and 18 patients in the delayed repair (DR) group repaired at 8.5 days (range, 17 hours 67 days). There were no significant differences between the ER and DR groups in age (37+/-18 years vs. 41+/-19 years), Injury Severity Score (39+/-15 vs. 45+/ 14), intensive care unit days (12+/-14 days vs. 18+/-11 days), hospital length of stay (21+/-19 days vs. 28+/-14 days), or mortality rates (7% vs. 6%). There was a trend toward longer lengths of stay in the DR group. Most DR patients required beta-blocker therapy and/or other antihypertensives for systolic BP more than 120 mm Hg during admission. There were no deaths from aortic rupture in either group. By using financial data that was available from July of 1994 onward, we performed a subset analysis of the direct costs associated with BTAI. Total direct and variable direct costs for patients undergoing delayed repair were over two times the costs for early repair patients (p < 0.05). CONCLUSION: The management of trauma patients with multiple injuries requires prioritization of injuries so that the outcomes from these injuries can be optimized. Although delayed aortic repair was safely practiced in this series, there was not an obvious outcome benefit to delayed repair. The patients undergoing late repair required increased attention to hemodynamics, and there was a trend toward increased length of stay. In addition, analysis of the costs associated with delayed repair demonstrated a twofold increase in the direct costs for delayed repair compared with early repair. PMID- 10452459 TI - Comparative study of splenic wound healing in young and adult rats. AB - BACKGROUND: Nonoperative management (NOM) of splenic injuries is a common practice in stable trauma patients. Nevertheless, age-related differences in the success rate of NOM have prompted inclusion of age among the criteria of patient selection. Elucidation of the cellular mechanism of splenic wound healing in the young versus that of adults may explain why age can be related to the success of NOM in splenic injuries. METHODS: A laceration was made in the splenic antihilar surface of 40 young and 40 adult male rats. Postoperatively, at specified intervals extending until day 21, spleens were removed, fixed, and examined by routine histopathology. In addition, sections were stained histochemically for collagen fibers and immunohistochemically for myofibroblast histomorphometry. RESULTS: The intense local hemorrhage was resorbed within 48 hours in the young rats, and within 7 days in the adults. Disappearance of germinal centers and other splenic alterations started on the first day in both groups, but regeneration of splenic parenchyma was accomplished after 14 days in the young, whereas in the adults, on day 21 it was still incomplete. Maximal myofibroblast accumulation at the laceration site was seen after two days in the young, whereas in adults only on day 4 (p < 0.0001). Collagen scars were not present in either group. Thickening of the damaged capsule, composed of collagen fibers with yellowish-green polarization colors, was observed only in adult rats. CONCLUSION: Splenic wounds heal by regeneration and not by collagen scarring. In the young, myofibroblasts accumulate in the site of injury faster than in adults. These cells may enhance contraction and increase the rate of wound healing until parenchymatic regeneration is completed. Our results may indirectly explain the higher success rate of NOM of splenic injury in young patients. PMID- 10452460 TI - Light deprivation soon after frontal brain trauma accelerates recovery from attentional deficits and promotes functional normalization of basal ganglia. AB - BACKGROUND: Light deprivation significantly accelerates recovery from attention deficits (neglect) after cortical ablation in rats. We hypothesized that light deprivation would improve recovery after traumatic contusive brain injury (TBI) and do so by enhancing dopaminergic function in the ipsilateral basal ganglia. METHODS: Adult rats received left frontal contusion injury and were placed into darkness or standard light/dark cycling for 48 hours. Neurologic evaluation included attentional and sensorimotor tasks. Amphetamine-induced production of the immediate early gene protein product Fos was quantified to determine neuronal dopaminergic response in caudate-putamen (striatum). RESULTS: Unilateral frontal TBI produced severe contralateral deficits in all tasks. Postoperative light deprivation resulted in improved recovery from attentional but not sensorimotor deficits. Five days after injury, ipsilateral striatal Fos expression was reduced by 51% in TBI rats experiencing normal light cycling (p < 0.006). In contrast, postoperative light deprivation normalized striatal Fos expression. By 6 weeks, all TBI rats demonstrated nearly full recovery and striatal Fos expression was symmetrical between the two striata. CONCLUSION: Postoperative light deprivation may improve recovery from TBI-induced attention deficits by normalizing basal ganglia function. PMID- 10452461 TI - Posttraumatic carotid cavernous fistula: frequency analysis of signs, symptoms, and disability outcomes after angiographic embolization. AB - OBJECTIVE: To increase awareness and understanding of posttraumatic carotid cavernous fistula (PTCCF) with the intent to expedite diagnosis and treatment of this disabling injury, a 14-year retrospective review of patients with angiographically identified PTCCF was conducted at this Level I trauma center. A frequency analysis of signs, symptoms, and disability was performed. The impact on disability of demographics, number of embolization attempts required for closure of the PTCCF, and time from injury to diagnosis was assessed by t test for independent samples. RESULTS: Nine patients were diagnosed with 10 PTCCFs. Mean patient age was 41.5 years. All patients with PTCCF had basilar skull fracture, loss of consciousness, bruit, and chemosis; 90% had exophthalmos; 70% had visual changes; 50% complained of headache; and 80% had some lasting disability. Mean age of patients with partial to total disability was 47 years, while the mean age of patients without lasting disability was 19.5 years (p = 0.013). No statistical correlation could be found between disability and sex, blunt versus penetrating injury, days to diagnosis, or number of embolization attempts. CONCLUSION: Patients sustaining head trauma with basilar skull fractures and presenting with the described signs and symptoms should be evaluated for PTCCF. Risk of disability does not appear to be influenced by number of attempts at embolization or time to diagnosis. However, age may have a significant impact on outcome. PMID- 10452462 TI - Vascular tone in patients with hemorrhagic shock. AB - BACKGROUND: In hemorrhagic shock, the alterations in arterial vascular tone, which are primarily regulated by adrenosympathetic influences are compensatory responses to bleeding. OBJECTIVE: The aim of this study was to evaluate vascular tone expressed by the volume elastic modulus (Ev) as a clinical monitor to detect the hypovolemic state. METHODS: Thirteen patients with hemorrhagic shock were studied. The initial Ev measurement was performed at arrival, and subsequent measurements were obtained 4 and 12 hours after arrival. Patients were divided into two groups by cluster analysis by using the Ev values at arrival and 4 hours after arrival. Circulatory parameters, the clinical course, and fluid were compared. RESULTS: The Ev values were identical at admission (cluster I vs. II: 456.4+/-197.1 vs 566.1+/-234.1 mm Hg, mean +/- SD). After 4 hours of fluid resuscitation, all patients were recovered from shock. In cluster I, the Ev remained high at 4 hours (523.4+/-75.1 mm Hg) and invasive treatments for hemostasis were required. In cluster II, the Ev significantly decreased at 4 hours (182.8+/-70.7 mm Hg, p < 0.01) and clinical courses were eventless thereafter. During 4 to 12 hours, more fluid was given in cluster I (p < 0.05). At 12 hours after arrival, the Ev values were identical in both clusters. CONCLUSION: These data demonstrate that the Ev increases in hemorrhagic shock. Furthermore, normotensive hypovolemic conditions generated by persistent bleeding can be detected by measuring the Ev. PMID- 10452463 TI - Thermal filament continuous thermodilution cardiac output delayed response limits its value during acute hemodynamic instability. AB - BACKGROUND: It has been suggested that measurement of continuous cardiac output (CCO) is an advancement in the management of critically ill patients. Our objective was to determine the accuracy of CCO during the rapid hemodynamic changes induced by hemorrhage and resuscitation. METHODS: In 12 anesthetized dogs (20.2+/-0.9 kg), pulmonary artery blood flow, our "gold standard" cardiac output, was measured with an sonographic flowprobe, whereas CCO, intermittent bolus cardiac output (ICO), and mixed venous oxygen saturation were measured with a thermodilution fiberoptic pulmonary artery catheter with a thermal filament. A graded hemorrhage (20 mL/min) was produced to a mean arterial pressure of 40 mm Hg, which was maintained at this level for 30 minutes. Total shed blood volume (701+/-53 mL) was retransfused at a rate of 40 mL/min, over 30 minutes, after which a massive hemorrhage (100 mL/min) was produced over 10 minutes. RESULTS: Hemorrhage induced significant decreases in mean arterial pressure, mixed venous oxygen saturation, and oxygen delivery, which were all restored during early resuscitation. However, CCO showed a delayed response after hemorrhage and resuscitation, compared with pulmonary blood flow, throughout the study (r = 0.549), matching only at baseline and at the end of both graded hemorrhage and resuscitation periods. There was a good correlation between ICO and pulmonary artery blood flow (r = 0.964) and no significant differences between them throughout the study. CONCLUSION: CCO has a delayed response during acute hemodynamic changes induced by hemorrhage and resuscitation. When sudden changes in mean arterial pressure or in mixed venous oxygen saturation are detected, cardiac output must be estimated by the standard bolus thermodilution technique, not by CCO. PMID- 10452464 TI - Direct current reduces accumulation of Evans Blue albumin in full-thickness burns. AB - BACKGROUND: Application of direct current (DC) to a burn wound limits extension of the zone-of-stasis and reduces wound tissue edema. OBJECTIVE: To study the effects of DC on extravasation of plasma proteins after burn by using Evans blue (EB) as a marker of plasma albumin. MATERIALS AND METHODS: Male Sprague-Dawley rats with 20% total body surface area full-thickness scalds (100 degrees C/10 sec) were used as the experimental model. Burn wounds were treated with plain nylon, silver-nylon, silver-nylon and 40 microA DC, or no dressing. EB (30 mg/kg) was injected immediately or at variably delayed postburn (PB) times and accompanied by DC application at various time intervals PB. Tissue content of Evans blue was assessed at different times after injection of the dye or infliction of burn injury. RESULTS: Evans blue albumin (EBA) concentration in untreated burn wounds (307.7 microg/g tissue) was nine times greater than in unburned skin (36.5 microg/g tissue) at 48 hours PB. When animals received a DC and EB injection immediately PB, DC treatment reduced EBA concentration by 60% at any time point PB. When EB was injected immediately PB, or at variably delayed times PB, accompanied by DC immediately PB, or at variably delayed times PB, DC reduced EBA accumulation at all examined times PB by more the 50% (p < 0.001). CONCLUSION: EBA and edema fluid accumulation in burn wound change in concert after injury and show similar response to DC treatment. PMID- 10452465 TI - Infradiaphragmatic central venous pressures reflect supradiaphragmatic pressures in stable burn patients. AB - OBJECTIVE: Because the skin of the groin is often spared, femoral central venous catheters are sometimes used in patients with extensive burns. The accuracy of central venous pressures obtained from the infradiaphragmatic location relative to the traditional supradiaphragmatic value is not known in this population. METHODS: Seventeen seriously injured but hemodynamically stable burn patients were enrolled in a protocol approved by the human studies committee in which, during scheduled central venous line rotations, supradiaphragmatic and infradiaphragmatic central venous pressures were determined simultaneously. RESULTS: These 17 patients were an average of 21.8+/-20.1 years old (range, 6 months to 61 years), and the average burn size was 60.8+/-22.6% (range, 20 to 90%). Supradiaphragmatic and infradiaphragmatic pressures correlated well, with an r value of 0.903, p less than 0.01. CONCLUSION: In the absence of clinically significant abdominal distention, infradiaphragmatic central venous pressure is an accurate reflection of supradiaphragmatic pressure, and indirectly, circulating blood volume. PMID- 10452466 TI - Polylactide bioabsorbable polymers for guided tissue regeneration. AB - OBJECTIVE: Guided tissue regeneration is a procedure to improve tissue repair, which creates an optimal environment for the intrinsic growth ability of tissues. METHODS: A prerequisite for guided tissue regeneration is the availability of materials with suitable physicochemical and biocompatibility properties for the preparation of the devices. We investigated bone and peripheral nerve guided tissue regeneration, making two conduits from poly[L-lactide-co-6-caprolactone] (PLLC--peripheral nerve) and with poly [DL-lactide] (PDLLA--bone) with different features. After the polymer synthesis and chemical characterization, the conduits were evaluated in vivo in rat sciatic nerve gaps and in rabbit radius defects. RESULTS: The results demonstrated good biocompatibility of both polymeric conduits. A good axonal regeneration and the restoration of the nerve trunk continuity, similar to that observed with autologous grafts has been obtained with PLLC conduits, that slowly degrade in about 6 months. PDLLA conduits protected the bone defect against the invasion of surrounding soft tissues; an effective bone growth bridging the defect was observed in their lumen. CONCLUSION: These results confirm the versatility of polylactides as biomaterials and will encourage further investigations on hard and soft tissues. PMID- 10452467 TI - Abbreviated injury scale unification: the case for a unified injury system for global use. AB - BACKGROUND: The Abbreviated Injury Scale (AIS), developed by the Association for the Advancement of Automotive Medicine is the most widely used anatomic injury severity scale in the world (Association for the Advancement of Automotive Medicine. The Abbreviated Injury Scale; 1985 and 1990 revisions. Des Plaines, IL: Association for the Advancement of Automotive Medicine). However, different user groups have modified the AIS system to fit their needs, and these modifications prevent ready comparison and trending of data collected in these systems in the United States and throughout the world. The United States currently has five AIS based severity systems and two AIS-based impairment systems in use, with additional revisions forthcoming. Other modified AIS systems are known to be in use in the United Kingdom and Japan. The data collected in these systems cannot be accurately combined or compared without re-coding or the use of complex "mapping" methodologies. Furthermore, the expanding use of data linked from multiple databases to answer complex medical, engineering, or policy issues emphasizes the need for coordination between severity and other injury systems. Linkage of state-wide motor vehicle crash data with data from hospital injury classification systems, mortality files, trauma registry, and national crash databases brings into immediate focus the lack of well defined relationships between the severity coding systems and these other widely used injury systems (Mango N, Garthe E. SAE Congress, February, 1998; Johnson, S, Walker, J. NHTSA Technical Report. DOT HS 808 338, Washington, DC: NHTSA; January, 1996). With the expanding use of linked data in state and national policy decisions, it is vital that consistent standards for injury descriptions, severities, and impairments be available for clinical, engineering, and policy users. METHODS: This paper compares five anatomic severity systems and two impairment systems in terms of purpose, code structure, and use and discusses the reasons for the differences between these systems. With global "harmonization" encouraging greater sharing of international data, the paper also presents the relationship of the severity and impairment systems to US morbidity and reimbursement and worldwide mortality classification systems. RESULTS: To resolve compatibility issues resulting from multiple injury systems, the authors propose that a "unified" system for global use be developed, configured by inputs from major "data owners," users, and analysts. The proposed unified system has six key attributes: backward compatibility with historical data through "maps" so no data are lost; "scalability" to allow a simple level of use for developing countries, a more complex level for crash research and a detailed level for clinical hospital use, all with data compatibility; the ability to satisfy the needs of the engineering community for injury location information and aspect, and also the clinical requirement for precise injury description; inherent integration with whole body severity scores to permit easy computation; compatibility with other injury data systems such as mortality, morbidity, and reimbursement systems; and a mechanism and process to maintain and upgrade the system into the 21st century. CONCLUSION: The authors believe that a "unified" injury system is a necessary and crucial advance from the currently fragmented injury system situation. Unified data can provide a pool of consistent international data to support a variety of important research and prevention and treatment efforts and is essential to satisfy the global needs of the medical and engineering communities. PMID- 10452468 TI - Prediction of outcome in intensive care unit trauma patients: a multicenter study of Acute Physiology and Chronic Health Evaluation (APACHE), Trauma and Injury Severity Score (TRISS), and a 24-hour intensive care unit (ICU) point system. AB - OBJECTIVE: To conduct a multicenter study to validate the accuracy of the Acute Physiology and Chronic Health Evaluation (APACHE) II system, APACHE III system, Trauma and Injury Severity Score (TRISS) methodology, and a 24-hour intensive care unit (ICU) point system for prediction of mortality in ICU trauma patient admissions. METHODS: The study population consisted of retrospectively identified, consecutive ICU trauma admissions (n = 2,414) from six Level I trauma centers. Probabilities of death were calculated by using logistic regression analysis. The predictive power of each system was evaluated by using decision matrix analysis to compare observed and predicted outcomes with a decision criterion of 0.50 for risk of hospital death. The Youden Index (YI) was used to compare the proportion of patients correctly classified by each system. Measures of model calibration were based on goodness-of-fit testing (Hosmer-Lemeshow statistic less than 15.5) and model discrimination were based on the area under the receiver operating characteristic curve (AUC). RESULTS: Overall, APACHE II (sensitivity, 38%; specificity, 99%; YI, 37%; H-L statistic, 92.6; AUC, 0.87) and TRISS (sensitivity, 52%; specificity, 94%; YI, 46%; H-L statistic, 228.1; AUC, 0.82) were poor predictors of aggregate mortality, because they did not meet the acceptable thresholds for both model calibration and discrimination. APACHE III (sensitivity, 60%; specificity, 98%; YI, 58%; H-L statistic, 7.0; AUC, 0.89) was comparable to the 24-hour ICU point system (sensitivity, 51%; specificity, 98%; YI, 50%; H-L statistic, 14.7; AUC, 0.89) with both systems showing strong agreement between the observed and predicted outcomes based on acceptable thresholds for both model calibration and discrimination. The APACHE III system significantly improved upon APACHE II for estimating risk of death in ICU trauma patients (p < 0.001). Compared with the overall performance, for the subset of patients with nonoperative head trauma, the percentage correctly classified was decreased to 46% for APACHE II; increased to 71% for APACHE III (p < 0.001 vs. APACHE II); increased to 59% for TRISS; and increased to 62% for 24-hour ICU points. For operative head trauma, the percentage correctly classified was increased to 60% for APACHE II; increased to 61% for APACHE III; decreased to 43% for TRISS (p < 0.004 vs. APACHE III); and increased to 54% for 24-hour ICU points. For patients without head injuries, all of the systems were unreliable and considerably underestimated the risk of death. The percentage of nonoperative and operative patients without head trauma who were correctly classified was decreased, respectively, to 26% and 30% for APACHE II; 33% and 29% for APACHE III; 33% and 19% for TRISS; 20% and 23% for 24-hour ICU points. CONCLUSION: For the overall estimation of aggregate ICU mortality, the APACHE III system was the most reliable; however, performance was most accurate for subsets of patients with head trauma. The 24-hour ICU point system also demonstrated acceptable overall performance with improved performance for patients with head trauma. Overall, APACHE II and TRISS did not meet acceptable thresholds of performance. When estimating ICU mortality for subsets of patients without head trauma, none of these systems had an acceptable level of performance. Further multicenter studies aimed at developing better outcome prediction models for patients without head injuries are warranted, which would allow trauma care providers to set uniform standards for judging institutional performance. PMID- 10452469 TI - Prognostic factors and management of civilian penetrating duodenal trauma. AB - OBJECTIVE: This study was designed to investigate risk factors in the final outcome of patients with civilian penetrating abdominal trauma and duodenal injuries, the value of the different surgical approaches used, and to define when more complex procedures are indicated, instead of the simple primary repair. METHODS: The study design was a retrospective review of prospectively collected data of a 4-year period (July 1992 to June 1996). RESULTS: A total of 167 patients were admitted with penetrating abdominal trauma and duodenal injuries at San Juan de Dios Hospital in Santafe de Bogota, Colombia. CONCLUSION: The independent and significant risk factors that determine the severity of duodenal injury and need for complex procedures, as identified in this series, are preoperative or intraoperative shock; Abdominal Trauma Index higher than 25; and associated injuries to the pancreas, superior mesentric vessels, and colon. These factors are associated with an increased incidence of septic complications, duodenal fistula, and late mortality. PMID- 10452470 TI - A randomized prospective trial of amphotericin B lipid emulsion versus dextrose colloidal solution in critically ill patients. AB - BACKGROUND: Amphotericin B is the agent of choice for most invasive fungal infections in critically ill patients. It is associated with at least a 50% incidence of nephrotoxicity, despite prophylactic measures such as sodium loading. Newer formulations of amphotericin B are available but are costly and have unknown bioavailability in critically ill patients. Previous trials in neutropenic and critically ill patients have demonstrated that mixing amphotericin B with 20% lipid solution (Intralipid; Clintec Nutrition, Deerfield, III) may decrease nephrotoxicity. METHODS: In this randomized, prospective clinical trial, patients with positive fungal blood cultures, tracheal/sputum cultures or peritoneal cavity cultures were randomized to receive either 0.5 mg/kg per day of amphotericin B dextrose or 1.0 mg/kg per day of amphotericin B lipid emulsion. Duration of therapy was determined by the primary care team. Weekly 24-hour creatinine clearance was measured until 2 weeks after amphotericin B therapy was completed. RESULTS: The two groups were similar based on age, white blood cell count, serum creatinine, and creatinine clearance at the beginning of therapy. The group receiving amphotericin B lipid emulsion had significantly less decrease in creatinine clearance compared with controls, despite receiving significantly more amphotericin B. CONCLUSION: Amphotericin B lipid emulsion can be given at a higher total cumulative dose than amphotericin B dextrose with less nephrotoxicity. PMID- 10452472 TI - Effect of alcohol intoxication on hemodynamic physiology and outcome in patients with traumatic cardiac tamponade. AB - BACKGROUND: Alcohol intoxication has a detrimental effect on hypovolemic shock. Our aim, was to study its effects on "pure" cardiac tamponade (i.e., without hypovolemia) in patients with penetrating chest injuries. METHODS: Thirty-five intoxicated and 15 nonintoxicated patients (blood alcohol > and < 17 mmol/L) were studied. Initial vital signs (trauma scores), special investigations (hematologic profiles, blood gases, glucose, lactate, and catecholamines), clinical progress (24- and 72-hour acute physiology and chronic health evaluation II scores) and outcome were compared. RESULTS: Intoxicated patients were older (p = 0.02) and more tachypneic on admission (p = 0.006), but no other differences were noted. Mortality was proportional to the degree of shock and was greater in patients who had "front-room" thoracotomies (p < 0.001). Despite the higher percentage of intoxicated patients who were "lifeless" or "in extremis" on admission, they fared no worse than nonintoxicated patients. CONCLUSION: Alcohol intoxication does not have an adverse affect on traumatic cardiac tamponade. PMID- 10452471 TI - Measurement of muscle protein synthesis by positron emission tomography with L [methyl-11C]methionine: effects of transamination and transmethylation. AB - BACKGROUND: Positron emission tomography with L-[methyl-11C]methionine provides a measure of regional protein synthesis rate (PSR) in skeletal muscle. However, the validity of the method depends on incorporation of methionine into protein with minimal transamination, transmethylation, or both. To test directly these assumptions, uptake of L-[methyl-14C]methionine in skeletal muscle was measured in control and cycloheximide-treated rats. METHODS: Normal and cycloheximide treated rats (n = 8/group) were injected with 50 microCi of L-[methyl 14C]methionine and arterial blood sampled over 90 minutes. After killing, thigh muscle was homogenized, centrifuged, and treated with trichloroacetic acid. PSR from circulating methionine was estimated from trichloroacetic acid-precipitable radioactivity, arterial time-activity curves, and plasma methionine concentrations. RESULTS: In normal rats, approximately 70% of the tissue radioactivity was precipitated with trichloroacetic acid. In normal animals, PSR was 0.22 nmoles x min(-1) x g(-1), in excellent agreement with previous results. In the cycloheximide-treated group, PSR was 0.0032 nmoles x min(-1) x g(-1); approximately 98% reduction compared with controls. CONCLUSION: These studies support the hypothesis that L-[methyl-11(14C]methionine accumulates in skeletal muscle as 11(14)C-labeled protein. PMID- 10452473 TI - Use of the laryngeal mask airway in air transport when intubation fails. AB - BACKGROUND: A prospective, nonrandomized cohort study was conducted to determine the effectiveness of the laryngeal mask airway (LMA) for management of the difficult airway in patients requiring air transport. METHODS: The LMA was inserted in those patients who could not be successfully intubated. Data were collected to evaluate the effectiveness of the LMA and to document any complications attributed to its use. RESULTS: Inclusion criteria were met in 17 of the 25 patients receiving an LMA. The device was inserted successfully in 16 of 17 of the patients (94%). In-flight oxygen saturation ranged from 97 to 100%, and end-tidal carbon dioxide ranged from 24 to 35 mm Hg. At arrival, initial arterial blood gas values indicated adequate oxygenation in all patients and adequate ventilation in 15 of 16 patients (94%). There was no evidence of complications. CONCLUSION: Our patient data show that when conventional methods have failed, the LMA can be safely, rapidly, and effectively used for temporary airway control. PMID- 10452474 TI - Early biochemical characterization of soft-tissue trauma and fracture trauma. AB - OBJECTIVE: The long-term outcome of trauma patients basically depends on the relation between the clearance capacity of the organism, e.g., the lungs, and the antigenic (inflammatory) load in relation to the amount of damaged and perfused tissue. It is necessary to determine quality and quantity of fracture and soft tissue damage by clinical means as early as possible. It is unknown whether biochemical markers and the impact of soft-tissue trauma correlate and whether there is a predictive value on clinical outcome. METHODS: A total of 107 trauma patients were prospectively enrolled in the study. Blood samples were collected immediately at the site of accident, at hospital admission, and every 2 hours for an interval of 24 hours, then daily. In addition to the biochemical analysis of 20 different substances, the following data were collected and correlated to the laboratory results: Injury Severity Score, polytrauma score of Hannover, modified fracture index, and soft-tissue index. These primary clinical findings as well as the laboratory data were correlated to criteria of clinical outcome such as length of stay in the intensive care unit, length of hospital stay, infections, systemic inflammatory response syndrome, sepsis, multiple organ failure score according to Goris, and finally to primary (< 72 hours), secondary (> 72 hours), and overall lethality. The determination of individual extent and severity of soft-tissue trauma is based on standard partial body volumes derived from healthy volunteers. In addition, clinical estimation of the degree of soft-tissue damage according to the usual classifications was performed. RESULTS: Significant (p > 0.05) correlations were found between fracture as well as soft-tissue trauma and intensive care unit stay, hospital stay, infections, systemic inflammatory response syndrome, multiple organ failure score, serum concentrations/activities of serum interleukin-6 and -8 and creatine kinase during the first 24 hours after trauma. Severe soft-tissue trauma was related to secondary lethality, however, without statistical significance. CONCLUSION: The amount of fracture and soft tissue damage can be estimated early by analysis of serum interleukin-6 and creatine kinase and is of great importance with regard to long-term outcome after trauma. PMID- 10452475 TI - Functional outcome of internal fixation for pelvic ring fractures. AB - OBJECTIVE: Evaluation of the functional outcome after unstable pelvic ring fractures stabilized with internal fixation. METHODS: Between January 1, 1990, and September 1, 1997, 37 patients were treated with internal fixation for unstable pelvic fracture. Demographic data, type of accident, Hospital Trauma Index-Injury Severity Score, and fracture type according to Tile classification were scored. One patient died the day after the accident from neurologic injury. A Short Form-36 health questionnaire and a form regarding functional result after pelvic trauma, adapted from Majeed et al., were returned by 31 of 36 patients (86%). Twenty-eight patients (78%) were seen for physical and radiologic examination. RESULTS: Twenty-six men and 11 women, with an average age of 34.7 years (range, 15-66 years) were included. The mean Injury Severity Score reached 30.4 (range, 16-66). According to the Tile classification, there were 16 type B fractures and 21 type C fractures. Seven patients were treated with open reduction and internal fixation of the pubic arch, 10 patients were treated with a combination of anterior open reduction and internal fixation with additional external fixation to increase the stability of the posterior ring. Nineteen patients underwent internal fixation of both anterior and posterior arch. In the remaining case, percutaneous posterior screw fixation was combined with anterior external fixation, because of estimated infectious risk. The average follow-up time was 35.6 months. Patients scored 78.6 of 100 on the Majeed score. Remarkable was the reported change in sexual intercourse in 12 patients (40%). Only 12 patients (40%) did not have complaints when sitting. On the SF-36 scales physical and social functioning, role limitations due to physical problems and vitality were limited compared with the averages for the Dutch population. Patients treated with combined anterior and posterior internal fixation scored significantly better on both the Majeed score and on the categories physical functioning, pain, general health and social functioning compared with patients with similar fractures treated with a combination of anterior internal fixation with external fixation. At the physical examination, 11 of 28 patients (39%) did not have any abnormality. Nineteen patients (68%) were back at their original job, which was physically demanding in 9 cases. There was a suspicion of nonunion of the posterior arch in two patients, which could be confirmed with a computed tomographic scan. CONCLUSION: In general, limitations in functioning are reported, even after long-term follow-up. In partially unstable fractures, solitary anterior fixation gives good results. In completely unstable fractures, patients treated with combined internal fixation anterior as well as posterior scored a better outcome compared with combined internal and external fixation. Therefore, this technique is recommended as treatment of first choice in completely unstable fractures. PMID- 10452476 TI - Elastic stable intramedullary nailing for unstable femoral fractures in children: preliminary results of a new method. AB - BACKGROUND: Many femoral fracture patterns in children cannot be stabilized sufficiently by intramedullary nailing only. Such fractures may require additional cast bracing or cerclage wiring after nailing. To overcome this problem, pediatric Ender nails that can be interlocked were designed to achieve better fracture stabilization. METHOD: Seventeen children (age, 2.5 to 15 years) were treated with this method for unstable traumatic fractures of the femur. The average follow-up period was 11.8 months. RESULTS: All fractures healed within 4 weeks in the mean. There were no major complications. CONCLUSION: This new method prevents shortening and axial deviation of the fractured femur. Start of postoperative mobilization and increase of weight-bearing is mainly determined by the child. PMID- 10452477 TI - Fatigue strength of locking screws and prototypes used in small-diameter tibial nails: a biomechanical study. AB - BACKGROUND: One major problem with the use of small-diameter nails in the treatment of tibial fractures is the high rate of fatigue fractures of the locking screws. The objective of this study was to correlate such parameters as diameter of locking screws, influence of notch effect, and material with fatigue strength and fatigue limit. METHODS: In a biomechanical study, the mechanical properties of 11 different locking screws and screw designs to be used in solid tibial nails were tested under typical fatigue conditions. RESULTS: The results showed that an increase in the screw's diameter significantly increased the fatigue strength of the tested locking screws and prototypes (p < 0.0005). A 20% increase of diameter improved fatigue strength by 25 to 70%. Absence of thread at the location where the screw contacts the nail's aperture (where the main load is transmitted to the screw and where the screw, therefore, usually tends to break) may double the fatigue strength and fatigue limit by avoiding the negative notch effect of the screw's thread. It was further significant that titanium alloys had a higher fatigue strength compared with stainless steel alloys (p < 0.001). However, the fatigue strength of titanium implants is massively reduced by minimal damage to the surface. CONCLUSION: Optimizing the core diameter of the locking screws at the aperture of the solid tibial nail by using screws without a continuous thread generates a fatigue strength two to three times higher compared with screws with a continuous thread. Increasing fatigue strength of locking screws might decrease malunion in fractures stabilized by small-diameter nails. PMID- 10452478 TI - Rectal blowout by personal watercraft water jet: case report and review of literature. PMID- 10452479 TI - The displaced open acetabular fracture: treatment with immediate open reduction and internal fixation. PMID- 10452480 TI - Thigh compartment syndrome as a result of a false aneurysm of the profunda femoris artery complicating fixation of an intertrochanteric fracture. PMID- 10452481 TI - Posttraumatic pseudoaneurysm of the sphenopalatine artery presenting as persistent epistaxis: diagnosis and management. PMID- 10452482 TI - False aneurysm of the left common carotid artery 52 years after penetrating injury of the chest. PMID- 10452483 TI - Aortic injury resulting from attempted subclavian central venous catheter placement. PMID- 10452484 TI - Traumatic rupture of the descending thoracic aorta associated with severe pulmonary contusion: requirement of full cardiopulmonary bypass for life support during surgical repair. PMID- 10452485 TI - Liver laceration in the hemodynamically unstable post-cardiac massage patient: early recognition and management--case report. PMID- 10452486 TI - Gastric rupture resulting from blunt abdominal trauma and requiring gastric resection. PMID- 10452487 TI - Diaphragmatic paralysis following minor blunt trauma. PMID- 10452489 TI - Somnambulism and trauma: case report and short review of the literature. PMID- 10452488 TI - Simultaneous revascularization and soft tissue coverage in the traumatized upper extremity with a flow-through radial forearm free flap. PMID- 10452490 TI - Bronchial blocker placement through the lumen of an in situ tracheal tube. AB - Tracheal intubation during trauma resuscitation is almost always performed with a standard endotracheal tube. Difficulties may arise if lung isolation is required later. The options for achieving lung isolation in seriously traumatized patients are briefly reviewed. Two efficient and airtight systems to allow the insertion of a bronchial blocker with minimal risks are presented. PMID- 10452491 TI - Hyperbaric oxygen treatment: a brief overview of a controversial topic. PMID- 10452492 TI - Trauma care economics: black, like "in the black". PMID- 10452493 TI - Demographics are an important aspect of any study. PMID- 10452494 TI - Seat belt syndrome. PMID- 10452495 TI - Temporary vena caval interruption device. PMID- 10452496 TI - Intracranial pressure monitor placement by midlevel practitioners. PMID- 10452497 TI - Patients with blunt carotid and vertebral artery injuries. PMID- 10452498 TI - Determining the diagnosis and prognosis of thyroid neoplasms: do special studies help? PMID- 10452499 TI - Recommendations for reporting of tumors of the adrenal cortex and medulla. Association of Directors of Anatomic and Surgical Pathology. AB - The Association of Directors of Anatomic and Surgical Pathology has developed recommendations for the surgical pathology report for common malignant tumors. The recommendations for tumors of the adrenal are reported herein. PMID- 10452500 TI - Expression of vascular endothelial growth factor (VEGF) in human thyroid neoplasms. AB - Vascular endothelial growth factor (VEGF), also known as vascular permeability factor (VPF), is an angiogenic factor that plays important roles in tumor growth. Angiogenesis studies on VEGF deal with various types of malignant tumors, but very little is known about the role or significance of VEGF in human thyroid neoplasms. Therefore, in the current study, we determined whether VEGF is found in normal and neoplastic thyroids and whether its expression is altered in different histological types of thyroid neoplasms. Reverse-transcription polymerase chain reaction (RT-PCR) analysis showed that all specimens of thyroid tumors expressed bands corresponding to 121-, 165-, and 189-amino acid forms of VEGF. Northern blot analysis showed an increase in VEGF mRNA levels in neoplastic tissues in comparison with normal thyroid samples. By nonisotopic in situ hybridization, most of the tumor cells in follicular adenomas expressed VEGF mRNA, whereas VEGF mRNA expression was identified only in epithelium of isolated follicles in normal thyroid tissues. In papillary thyroid carcinomas, an intense labeling with VEGF probe was often found in overlying tumor cells of neoplastic papillae. VEGF expression was distinctly intensified in undifferentiated carcinoma cells that were immediately adjacent to necrotic foci. The immunohistochemical localizations of VEGF protein were comparable to the localization of VEGF mRNA. In conclusion, our results suggest that the histological types of thyroid tumor may determine the vascular pattern through a paracrine mechanism involving VEGF. PMID- 10452501 TI - The differentiation of true adenomas from colitis-associated dysplasia in ulcerative colitis: a comparative immunohistochemical study. AB - Adenomas in areas involved by ulcerative colitis (UCA) are difficult to identify because of their morphological similarity to ulcerative colitis-associated dysplasia (UCD) and have an uncertain biology. Recently, a set of morphopathologic criteria were published for the diagnosis of UCA versus UCD. As a first step to analyze these criteria, we studied p53 and bcl-2 expression in groups of UCA and UCD along with a sporadic adenoma control group. Ninety lesions from UC areas (62 patients) were examined, including 24 UCA without high-grade dysplasia (HGD) and 66 UCD consisting of 43 polypoid and 23 flat dysplastic lesions (29 with HGD). Immunohistochemical p53 and bcl-2 expression were evaluated semiquantitatively. P53-positive cases were significantly less frequent in the UCA (4%) versus the UCD group (30%, P = .01) and the polypoid UCD subgroup (35%, P = .005). Moderate or strong bcl-2 expression was significantly more frequent in the UCA than in the UCD group (96% v 70%, P = .01) and in the UCA versus both polypoid and flat UCD subgroups. Comparison of UCA with low-grade dysplastic polypoid UCD cases alone showed a difference just below significance for p53 (P = .07). p53 and bcl-2 expression rates were very similar in the UCA group and the sporadic adenoma (n = 25) control group. These results show that UCA has phenotypic features more similar to sporadic adenomas than UCD and supports the concept that adenomas in UC have a biology different from UC associated dysplasia. PMID- 10452502 TI - Loss of retinoblastoma protein expression is frequent in small cell neuroendocrine carcinoma of the cervix and is unrelated to HPV type. AB - We have previously identified an inverse relationship between p53 and retinoblastoma protein (pRb) immunoreactivity in non-small cell carcinoma of the cervix. Because pRb is infrequently expressed in small cell carcinoma of the lung, we analyzed 25 small cell neuroendocrine carcinomas of the cervix to test the hypotheses that 1) lack of pRb expression is associated with the neuroendocrine phenotype in human papillomavirus (HPV)-associated cervical carcinoma and 2) the inverse relationship between p53 and pRb immunoreactivity also occurs in these tumors. HPV type was analyzed by PCR, HPV distribution by in situ hybridization and expression of p53 and pRb by immunohistochemistry. All of the tumors contained HPV sequences, with 13 tumors HPV 16 positive, 11 HPV 18 positive, and 1 HPV 45 positive. In situ hybridization showed large intranuclear dot-like signals in all positive tumors, suggesting viral integration. No multiple infections were identified. Expression of retinoblastoma protein was not detectable in 23 tumors (92%), the remaining two showing only weak, focal expression. Expression of p53 protein was variable in distribution and intensity. It did not correlate with HPV type, and there was no relationship with pRb immunoreactivity. These data indicate that, although there is no reciprocal relationship between p53 and pRb immunoreactivity in these tumors, retinoblastoma protein is infrequently expressed in HPV-containing small cell neuroendocrine carcinoma of the cervix, irrespective of infecting HPV type. This is consistent with the reported findings in small cell carcinoma of the lung and suggests that the small cell neuroendocrine phenotype may be related to the abrogation of retinoblastoma protein function. PMID- 10452503 TI - Neuroectodermal differentiation in Ewing's sarcoma family of tumors does not predict tumor behavior. AB - The observation that neuroectodermal differentiation imparts a worse prognosis to the Ewing family of tumors has been suggested by some studies and refuted by others. To assess whether the diagnosis of Ewing's sarcoma versus peripheral primitive neuroectodermal tumor (PNET) affects prognosis, we analyzed tumors from 63 analogously treated pediatric and young adult patients from the National Cancer Institute and St Jude Children's Research Hospital and retrospectively compared the results with clinical outcomes. The tumors were assessed using standard light microscopy and immunohistochemical stains for neuron-specific enolase, CD57, S100 protein, neurofilament protein, and synaptophysin with or without antigen retrieval. Ultrastructural evaluation was also performed in 39 tumors. Classification was performed using Kiel criteria as well as a modified classification. Kaplan-Meier analyses, with Mantel-Haenzel evaluation of the significance of the differences, were performed separately for localized or metastatic tumors. Using the Kiel classification on a subset of 60 cases, 39 tumors qualified as PNET and 21 as Ewing's sarcoma. Using the modified classification on a subset of 61 cases, 14 were classified as PNET, 21 as atypical Ewing's sarcoma, and 26 as Ewing's sarcoma. The addition of electron microscopy to the diagnostic armamentarium significantly increased the likelihood of identifying PNET. No significant differences in event-free or overall survival were seen using either the modified or Kiel classification, regardless of the ancillary diagnostic techniques employed. In this exploratory analysis, neuroectodermal differentiation did not play a role in clinical outcome. Confirmation of this finding will require a larger, separate study of similarly treated patients, and it may not apply to older patients. PMID- 10452504 TI - Neovascularization in early atherosclerotic lesions of human carotid arteries: its potential contribution to plaque development. AB - Neovascularization is a prominent feature of late-stage atherosclerotic lesions and their complications but is generally regarded as an insignificant, undetectable component of the earliest stages of plaque development, probably because of relatively poor histological techniques. Using an improved vascular staining procedure, we have examined the extent of neovascularization in the earliest plaque lesions. Combined monoclonal antibodies to CD31, CD34, and von Willebrand factor have provided an ultrasensitive technique with which to visualize blood vessels in early atherosclerotic lesions (n = 55) of human carotid arteries obtained through surgical procedures. Capillary-like microvessels were shown in very early atherosclerotic lesions (type II), where they were associated with the distribution of macrophages and a few immature mast cells. Neovascularization was more prominent in type III lesions with vessels of variable size, often providing a focus around which local accumulations of macrophages and apolipoproteins A-I and B were visualized. Thickened type III lesions usually showed an intricate network of microvessels, together with numerous mast cells. These studies have shown neovascularization as a prominent feature of early stages of atherosclerotic plaque development. Whereas distribution of apolipoproteins A-I and B were observed in the very earliest stages of the plaque intima, these lipids, together with macrophages, foam cells, and mast cells, were observed as perivascular accumulations in a proportion of type II and III lesions. Such findings indicate that neovascularization is an important feature of early plaque development and may provide an additional or alternative source of leukocyte and lipid accumulations relative to the arterial lumen. PMID- 10452507 TI - Comparison of pathological and biological features of symptomatic and mammographically detected ductal carcinoma in situ of the breast. AB - To determine whether the ductal carcinoma in situ (DCIS) detected mammographically or presenting clinically is the same or differs, pathological and biological (c-erbB-2 and p53 detection) features of 79 cases of pure DCIS, 5 cases with microinvasion and 8 cases with 1 to 2 mm of invasion, all detected by mammography, have been compared with 59 cases of pure DCIS, 8 cases with microinvasion and 7 cases with 1 to 2 mm invasion, all of which presented clinically. Half of the mammographically detected group were smaller than 20 mm, and there was a higher incidence of these being low grade, whereas 30% of the symptomatic cases were smaller than 20 mm, and more of this group were larger than 50 mm. For the pure DCIS, there were less high-grade and more intermediate grade cases in the mammographically detected group, although the incidence of low grade was similar between the two groups. There were more cases with a micropapillary pattern in the symptomatic group. C-erbB-2 protein was detected in 42% of the mammographically detected cases, whereas 59% of the symptomatic cases had c-erbB-2 reactivity. P53 detection was similar for both groups (33.0% and 37.0%). There were more symptomatic cases with invasion, and these were predominantly high grade, whereas the mammographically detected cases were both high and intermediate grade. Twelve of the 15 symptomatic cases with invasion expressed c-erbB-2, in comparison with 4 of the 13 mammographically detected cases, with half of the high-grade lesions in the latter group being negative. This study has shown that although there is overlap of pathological and biological features between DCIS presenting clinically and that detected mammographically, there can be differences in extent, grade, and invasion. The impact of this, however, can be determined only by clinical follow-up. PMID- 10452505 TI - Immunoreactivity of p53, Ki-67, and Bcl-2 in oncocytic adenomas and carcinomas of the thyroid gland. AB - To analyze relevant factors of neoplastic transformation in oncocytic neoplasms of the thyroid, expression of p53, Ki-67, and bcl-2 has been studied in oncocytic carcinomas (n = 17) and compared with results obtained in oncocytic adenomas (n = 20). P53 protein accumulation was found immunohistochemically in 75% of the oncocytic adenomas (15 of 20) and 88% of the oncocytic carcinomas (15 of 17). Eight of 17 of the carcinomas (47%), but only 3 of the 20 adenomas (15%), showed nuclear p53 accumulation in more than 10% of the cells, mostly in a focal pattern. Ki-67 expression also differed significantly between adenomas and carcinomas. The median of Ki-67-positive cells was 12/10 high-power fields (HPF) for adenomas and 76/10 HPF for carcinomas (P < .001). Furthermore, metastatic carcinomas had a significantly higher Ki-67 positivity than nonmetastasized carcinomas (164/10 HPF v 42/10 HPF, P < .05). Bcl-2 immunohistochemistry showed a constantly positive reaction in normal thyroid tissue. In contrast, bcl-2 protein was not detected in most of the adenomas (70%) and carcinomas (76%). In conclusion, p53 protein and Ki-67 is more prevalent in oncocytic carcinomas than in oncocytic adenomas of the thyroid, indicating that these factors may be involved in the progression of oncocytic neoplasms in the thyroid. In contrast, loss of bcl-2 appears to be an early event in the formation of oncocytic neoplasms of the thyroid. Its importance for malignant transformation is, however, unclear. PMID- 10452506 TI - Epithelioid sarcoma: an immunohistochemical analysis of 112 classical and variant cases and a discussion of the differential diagnosis. AB - Epithelioid sarcoma (ES) is a distinctive soft tissue neoplasm with a predilection for the distal extremities of young adults. This tumor typically contains nodular aggregates of epithelioid and spindle cells with zonal necrosis. The neoplastic cells are generally reported to coexpress keratin and vimentin and are often stated to be positive for CD34. However, there is no large series with extensive immunohistochemical data, there are few data with regard to expression of different keratin subtypes, and there are no large series discussing the epithelioid sarcoma subtypes. In the current study, we immunohistochemically evaluated 88 typical and 24 variant (8 angiomatoid, 9 large cell/rhabdoid, and 7 "fibroma-like") ESs. Nearly all ESs with typical histology (94%) were positive for keratin 8 (K8), whereas 72% were positive for K19, 48% for intermediate- and high-molecular-weight keratins (34betaEH12), and 22% for K7; reactivity with the latter two antibodies was usually seen in only a minority of tumor cells. Vimentin reactivity was present in all cases, EMA in 96% of cases and muscle specific actin and CD34 were noted in 41% and 52% of the cases, respectively. A few ESs (7%) showed focal cytoplasmic CD31 reactivity, but none exhibited a distinctive membrane staining pattern, and examples tested for FVIIIRAg were negative. The angiomatoid, fibroma-like, and large cell-rhabdoid ES variants had immunohistochemical profiles similar to the classic cases, supporting a common pathogenesis. Although not consistently expressed in ES, the presence of CD34 is helpful in distinguishing this entity from primary and metastatic carcinomas and other sarcomas such as malignant rhabdoid tumor. PMID- 10452508 TI - Prognostic significance of a novel proliferation marker, anti-repp 86, for endometrial carcinoma: a multivariate study. AB - Traditional prognostic factors often fail to identify a subgroup of endometrial carcinoma (EC) patients with an apparently paradoxical poor outcome. We therefore analyzed tumor cell proliferation immunohistochemically in a series of 164 endometrial carcinomas (EC) and compared its prognostic impact with that of the standard prognostic factors patient age, FIGO stage, FIGO grading, and histopathologic subtype. In addition to the established proliferation markers Ki S5 (Ki-67) and KiS4 (topoisomerase IIalpha), we used a novel monoclonal antibody (MAb), anti-repp 86, which binds to a recently described proliferation-specific protein (p86) expressed exclusively in the S, G2, and M phases of the cell cycle. anti-repp 86, Ki-S4, and Ki-S5 immunoreactive labeling indices (LI) correlated significantly with FIGO stage, FIGO grade, and myometrial invasion, but not with histopathologic subtype. By univariate analysis, conventional prognostic factors and proliferation indices were all predictive of disease-related mortality. A multivariate Cox regression analysis selected anti-repp 86 LI (P = .002), FIGO stage (P = .02), and histopathologic type as significant prognosticators of recurrence; anti-repp 86 LI (P = .001) and histopathologic type (P = .0106) also emerged as relevant predictors of mortality. A hierarchical forward regression model with the conventional prognosticators entered first and with anti-repp 86 entered next showed that anti-repp 86 and histopathologic subtype were the superior independent prognostic indicators for an increased risk of recurrence and cancer-related death. We conclude that the evaluation of anti-repp 86 immunostaining is an easily performable and exceptionally reliable method for identifying EC patients with adverse prognosis. PMID- 10452509 TI - Medullary thyroid microcarcinoma: a clinicopathologic retrospective study of 38 patients with no prior familial disease. AB - Thirty-eight patients (25 women, 13 men; mean age, 57.8 [32 to 91]) showing one or more medullary thyroid microcarcinomas (ie, < 1 cm), with no prior MEN II or medullary thyroid carcinoma history in their family, were reviewed. Follow-up was available for 29 patients (mean, 53.6 months [1 to 147]). 21 patients (72.4%) are alive and free of disease, four patients (13.8%) died during follow-up without disease, 2 patients are alive with disease (local recurrence and persistent hypercalcitoninemia) after 80 and 99 months, respectively, and 2 patients died of disease after 24 and 46 months. Most tumors were incidental pathological findings (19 of 38) or were discovered by systematic blood calcitonin measurement for a nodular thyroid disease (15 of 38). Only the four patients who had an unfavorable outcome were symptomatic cases (palpable micro-MTC, diarrhea, cervical lymph node metastasis and pulmonary metastatic disease). The two patients with metastatic disease at diagnosis died during follow-up. In univariate analysis, a symptomatic medullary thyroid carcinoma was a strong predictor of an unfavourable outcome (p < .00008), as were the preoperative calcitonin level (P = .007) and an elevated postoperative calcitonin level (P = .004). Among 30 histopathological criteria, only the presence of amyloid correlated with an unfavorable outcome (P = .018). PMID- 10452510 TI - Laminin and collagen IV in pleomorphic adenoma and carcinoma ex-pleomorphic adenoma: an immunohistochemical study. AB - Laminin and collagen have been studied in several tumor types and their immunomorphological expression correlated with tumor morphogenesis, local invasiveness, and metastatic behavior. In a series of 53 cases of pleomorphic adenomas (PA) and 16 cases of carcinomas ex-pleomorphic adenoma (Ca ex-PA), we investigated by immunohistochemistry the distribution pattern of laminin (lam) and collagen IV (col IV) at the basement membrane (BM) and the interstitial compartments, and their potential relationship with the metastatic behavior of Ca ex-PA. At the BM compartment, lam and col IV were expressed in 37 and 41 of all cases, respectively. In PA, there were lam and col IV around cell aggregates in 47.2% and 56.6% of the cases, respectively, and around isolated cells in 30.2% and 26.4%, respectively. In Ca ex-PA cases, both antigens were expressed in 56.3% and in 18.8% of the cases around cell aggregates and isolated cells, respectively. At the interstitial compartment, the fibro-hyaline matrix contained few lam in PA (19.2%) in contrast to Ca ex-PA (75%), including the benign areas of the neoplasms (90%), being the difference statistically significant (P < .001). In the Ca ex-PA group, a statistically significant difference was found on col IV deposits around tumor cell aggregates between metastasizing and nonmetastasizing neoplasms (P < .001). These findings support that laminin and collagen IV are involved in the process of malignant transformation of pleomorphic adenomas and their biological progression. PMID- 10452511 TI - Human polyoma virus in renal allograft biopsies: morphological findings and correlation with urine cytology. AB - Human polyoma virus (PV) interstitial nephritis occurs in immunosuppressed patients after reactivation of latent virus in renal epithelium. Currently, there is neither general consensus about the incidence of clinically significant PV infection in renal transplants nor conclusive evidence determining its significance in the long-term graft outcome. We evaluated 601 renal transplant biopsy specimens (from 365 patients) by routine light microscopy and immunoperoxidase stains with antibody against SV40 (which cross reacts with PV). We also examined urine samples from 200 patients (100 obtained concurrently with a renal biopsy in patients presenting with acute graft dysfunction and 100 from patients with stable graft function). Electron microscopic evaluation was performed in 50 renal biopsy specimens and in 23% of all urine samples. PV was identified in 1.8% biopsy specimens (1.9% of patients). PV interstitial nephritis showed the typical viral cytopathic changes in tubular epithelial cells associated with marked tubular damage and a disproportionately mild degree of tubulitis. There was no difference in the incidence of PV in the urine of patients with acutely deteriorating versus stable renal function (18% and 19%, respectively); however, urines with large numbers of infected cells (> 10/cytospin) and inflammatory changes in the sediments corresponded invariably to patients with acute allograft dysfunction (8 of 8), and in most cases to biopsy specimens showing PV interstitial nephritis (7 of 8). Based on these findings, urine samples seem to be the most sensitive and cost-effective screening method for PV infection; only urine samples with inflamed sediments and abundant infected cells correlate with clinically significant disease. In these cases, examination of a renal biopsy is indicated. Immunohistochemical stains are useful to confirm the presence of PV but do not increase the sensitivity of diagnosis of PV if this is not already suspected on routine light microscopy. In our material, immunostains were helpful ruling out the presence of PV in a small number of biopsy specimens (2%) that showed markedly reactive tubular cells resembling PV infection. Most patients with PV interstitial nephritis responded to decreased immunosuppression; however, the decay in graft function (based on creatinine slopes) was significantly more rapid in these patients than in matched controls. Evidence of PV infection should be systematically sought in renal biopsy specimens and urine samples from renal allograft recipients. PMID- 10452512 TI - Primary anaplastic large cell lymphoma of the central nervous system. AB - Central nervous system (CNS) involvement is extremely rare in anaplastic large cell lymphoma (ALCL), and in children only isolated cases have been reported, mainly as secondary CNS involvement. A case of fatal primary ALCL of the brain in a 13-year-old white boy is reported. Magnetic resonance imaging of the brain showed decreased absorption in T1- and T2-weighted image showed a hyperintense signal in the right parietal lobe and 2 masses in the right frontal lobe. A frontal lobe biopsy showed a pleomorphic neoplasm diffusely infiltrating the brain parenchyma and composed of large cells with bizarre, often polylobated or horseshoe-shaped nuclei. Immunohistochemical stains showed diffuse strong positivity for CD30, anaplastic lymphoma kinase protein (ALK-1), p80, leucocyte common antigen, CD45RO (UCHL1), and focal staining for epithelial membrane antigen. Immunostainings for cytokeratins, monocyte-macrophage, and B-cell markers were negative. Epstein-Barr virus latent membrane protein was not detected. To the best of our knowledge, there is only 1 case of primary ALCL of the brain in childhood previously reported in the literature. Before the biopsy, both cases were clinically misdiagnosed as mycobacterial CNS infection. Therefore, primary ALCL should also be included in the differential diagnosis when a mycobacterial CNS infection is suspected in pediatric patients; a careful cytological evaluation of the cerebrospinal fluid or cerebral biopsy are essential for an accurate diagnosis. PMID- 10452513 TI - Kaposi's sarcoma-associated herpesvirus-positive primary effusion lymphoma arising in the subarachnoid space. AB - Primary effusion lymphoma (PEL) is a rare and distinctive type of B-cell non Hodgkin's lymphoma (NHL) that occurs primarily, although not exclusively, in patients with AIDS. It usually develops as a lymphomatous effusion in the absence of a tumor mass, characteristically contains the Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8), usually also contains the Epstein Barr virus (EBV), displays a characteristic cytomorphology bridging immunoblastic and anaplastic large cell lymphoma, often expresses an indeterminate immunophenotype, and a B-cell genotype. Thus far, PEL has been limited almost entirely to the pleural, peritoneal, and pericardial cavities. We describe a NHL occurring in a gay man with AIDS that is typical of PEL in that it arose in a body cavity or space without an associated tumor mass, displays the cytomorphology typical of PEL, is a clonal B-cell neoplasm, and contains KSHV as well as EBV. This case is singularly distinctive in that it is the first case of PEL reported to arise in the subarachnoid space. This unique case further supports the strong association between KSHV and malignant lymphoma arising in body cavities and growing as an effusion. PMID- 10452514 TI - Molecular analysis of malignant triton tumors. AB - Triton tumors are rare variants of malignant peripheral nerve sheath tumor (MPNST) with muscle differentiation, often seen in patients with neurofibromatosis 1 (NF1). Individuals affected with NF1 harbor mutations in the NF1 tumor suppressor gene and develop neurofibromas and MPNSTs. The NF1 gene is expressed in Schwann cells and its expression is lost in schwannian neoplasms, suggesting a role in malignant development. Separately, there is evidence that p53 suppressor gene mutations are involved in MPNSTs. To determine the role of the NF1 and p53 genes in the development of the malignant Triton tumor we examined 2 such tumors, 1 from a 3-year-old boy without clinical manifestations of NF1 and another from a 24-year-old man with NF1. Histological analysis of these tumors showed both neural and muscle differentiation with S-100 and desmin immunoreactivity, respectively. Reverse transcribed RNA polymerase chain reaction (RT-PCR) of NF1 mRNA showed NF1 expression in the sporadic tumor. Strong nuclear immunoreactivity for p53 was observed throughout the malignant population in both tumors. This was confirmed by loss of heterozygosity for p53 in the non-NF1 patient, suggesting that p53 is involved in both hereditary and sporadic Triton tumors. The finding of preserved NF1 gene expression in the non-NF1-related Triton tumor suggests that different genetic events predispose to the development of this rare neoplasm in sporadic cases. PMID- 10452515 TI - A distinctive composite lymphoma consisting of clonally related mantle cell lymphoma and follicle center cell lymphoma. AB - Although follicle center cell lymphoma and mantle cell lymphoma are both B cell non-Hodgkin's lymphomas (NHL), they are regarded as separate entities with distinct clinical, morphological, immunophenotypic and molecular characteristics. To our knowledge, the coexistence of these 2 lymphomas in the same patient has never been reported. We describe a 70-year-old woman with a long-standing history of follicle center cell lymphoma, cytological grade I, who subsequently developed a composite lymphoma consisting of well-demarcated foci of persistent follicle center cell lymphoma surrounded by mantle cell lymphoma. This morphological interpretation was supported by the presence of both bcl-1 and bcl-2 gene rearrangements, which are molecular genetic hallmarks of mantle cell lymphoma and follicle center cell lymphoma, respectively. Polymerase chain reaction (PCR) analysis for rearranged immunoglobulin heavy chain (IgH) genes showed a dominant band identical in size in microdissected tumor cells of the follicle center cell and mantle cell lymphomas. Cloning and sequence analysis of the PCR products revealed a common clone-specific IgH gene rearrangement in these 2 lymphomas. These findings suggest that this composite lymphoma represents the unusual evolution of a malignant B-cell clone that resulted in the development of 2 morphologically distinct but clonally related B-cell NHLs. These findings also show the importance of integrating morphological, immunophenotypic, and molecular data to enhance our understanding of the complex pathogenic interrelationships in lymphomagenesis. PMID- 10452516 TI - Increased transforming growth factor-beta2 expression in the glomerular arteriole of the juxtaglomerular apparatus in a Bartter's-like syndrome. AB - Although transforming growth factor-beta (TGF-beta) has been shown to participate in regulating hormone synthesis and release, little is known about involvement of individual human TGF-beta isoforms, TGF-beta1, -beta2, and -beta3, in renin synthesis and release. We examined expression of these TGF-beta isoforms in a 50 year-old man with a Bartter's-like syndrome whose renal biopsy specimen showed hyperplasia of the juxtaglomerular apparatus (GA), mild mesangial hypercellularity, focal tubular atrophy, and interstitial fibrosis. Immunoreactivity for renin and marked expression of TGF-beta2 mRNA were noted in the glomerular arteriole of the JGA, whereas mRNA expression for TGF-beta1 was only slight and that for TGF-beta3 was still more faint. Expression of mRNAs for all 3 TGF-beta isoforms was increased in the fibrotic interstitium. This expression pattern suggests that TGF-beta2 may be involved in inducing renin synthesis and/or release in the glomerular arteriole of the JGA. PMID- 10452517 TI - Atypical immature metaplasia of cervix. PMID- 10452518 TI - The neoxanthin binding site of the major light harvesting complex (LHCII) from higher plants. AB - The localisation of the xanthophyll neoxanthin within the structure of the major light harvesting complex (LHCII) of higher plants has been investigated by site directed mutagenesis and spectroscopic methods. Mutation analysis performed on pigment binding sites in different helix domains leads to selective loss of neoxanthin for mutations on helix C thus localising this pigment between the helix C and helix A/B domains. Recombinant proteins binding two lutein molecules per polypeptide but lacking neoxanthin have been used in order to determine the contribution of neoxanthin to the absorption and linear dichroism spectra. The data were used to derive the orientation of the neoxanthin transition moment, lying in the polyene chain, which was thus determined to form an angle of 57 +/- 1.5 degrees with respect to the normal to the membrane plane where the protein is inserted. On the basis of these results we propose a model for the localisation of the carotenoid site in the LHCII structure which is still unresolved. PMID- 10452519 TI - Molecular characterization and immunolocalization of Dictyostelium discoideum protein phosphatase 2A. AB - Protein phosphatase 2A (PP2A) was previously purified from Dictyostelium and biochemically characterized. The purified PP2A holoenzyme was composed of a 37 kDa catalytic 'C-subunit', a 65 kDa 'A-subunit' and a 55 kDa 'B-subunit'. We report here the characterization of the genes encoding the Dictyostelium PP2A subunits as well as the immunolocalization of the PP2A subunits in Dictyostelium. The cDNAs encoding the B- and C-subunits were isolated from a Dictyostelium library and the deduced amino acid sequences reveal strong conservation with the mammalian PP2A homologues. Southern blot analysis suggests that each of the PP2A subunit genes is present in a single copy. The PP2A subunits were localized mainly to the cytosol in Dictyostelium cells. However, immunofluorescence confocal microscopy demonstrates that the B-subunit of PP2A is highly enriched in centrosomes, suggesting a potential role for this PP2A regulatory subunit in the centrosomal function. PMID- 10452520 TI - Knock-out of the cyaY gene in Escherichia coli does not affect cellular iron content and sensitivity to oxidants. AB - Friedreich ataxia is a recessively inherited neurodegenerative disease caused by deficiency of a highly conserved mitochondrial protein, frataxin. Frataxin deficiency results in mitochondrial iron accumulation and oxidative stress. Frataxin shows homology with the CyaY proteins of gamma-purple bacteria, whose function is unknown. We knocked out the CyaY gene in Escherichia coli MM383 by homologous recombination and we generated an E. coli MM383 strain overexpressing CyaY. Bacterial growth, iron content and survival after exposure to H2O2 did not differ among these strains, suggesting that, despite structural similarities, cyaY proteins in bacteria may have a different function from frataxin homologues in mitochondria. PMID- 10452521 TI - Molecular cloning and functional expression of a human peptide methionine sulfoxide reductase (hMsrA). AB - Oxidation of methionine residues in proteins to methionine sulfoxide can be reversed by the enzyme peptide methionine sulfoxide reductase (MsrA, EC 1.8.4.6). We cloned the gene encoding a human homologue (hMsrA) of the enzyme, which has an 88% amino acid sequence identity to the bovine version (bMsrA). With dot blot analyses based on RNA from human tissues, expression of hMsrA was found in all tissues tested, with highest mRNA levels in adult kidney and cerebellum, followed by liver, heart ventricles, bone marrow and hippocampus. In fetal tissue, expression was highest in the liver. No expression of hmsrA was detected in leukemia and lymphoma cell lines. To test if hMsrA is functional in cells, we assayed its effect on the inactivation time course of the A-type potassium channel ShC/B since this channel property strongly depends on the oxidative state of a methionine residue in the N-terminal part of the polypeptide. Co-expression of ShC/B and hMsrA in Xenopus oocytes significantly accelerated inactivation, showing that the cloned enzyme is functional in an in vivo assay system. Furthermore, the activity of a purified glutathione-S-transferase-hMsrA fusion protein was demonstrated in vitro by measuring the reduction of [3H]N-acetyl methionine sulfoxide. PMID- 10452522 TI - Role of the 14-3-3 proteins in the regulation of H+-ATPase activity in the plasma membrane of suspension-cultured sugar beet cells under cold stress. AB - All higher plants possess highly specific binding sites for fusicoccin, a metabolite of the fungus Fusicoccum amygdali Del. These sites are harboured in the plasma membranes and formed by a 14-3-3 protein dimer associated with the C terminal autoinhibitory domain of H+-ATPase. We considered the fusicoccin binding to plasma membranes to be an indicator of complexation between the 14-3-3 dimer and H+-ATPase, we assessed the effect of cold stress on the interaction of these proteins in suspension-cultured sugar beet cells and protoplasts derived from these cells. In both objects, upon lowering the temperature to 0-4 degrees C, a portion of the cytoplasmic 14-3-3 proteins became associated with the plasma membrane, which showed an increasing amount of ATPase/14-3-3 complexes and enhanced ATPase activity. Association between ATPase and 14-3-3 resulted in a several-fold rise in the H+ efflux from protoplasts and intact cells. We suppose that regulation of the H+ pumping under changing external conditions may be based on the interaction between H+-ATPase and the 14-3-3 proteins. PMID- 10452523 TI - Comparison of the roles of the C1a and C1b domains of protein kinase C alpha in ligand induced translocation in NIH 3T3 cells. AB - To explore the relative roles of the two C1 domains of protein kinase C alpha (PKC alpha) in the response to phorbol esters and related analogs, we mutated the individual C1 domains, expressed the mutated PKC alpha in NIH 3T3 cells, and then examined the ability of ligands to induce its translocation to the membrane. The C1a and C1b domains play equivalent roles for translocation in response to phorbol 12-myristate 13-acetate, mezerein, and (-)octylindolactam V. These results contrast with those previously reported for PKC delta, suggesting that the domains play different roles in different PKC isoforms. PMID- 10452524 TI - The antiproliferative activity of the murine interferon-inducible Ifi 200 proteins depends on the presence of two 200 amino acid domains. AB - Interferon-inducible proteins, p200, have a modular organization consisting of one (p203) or two (p202 and p204) 200 amino acid motifs, designated as type a or b domains. The relationship between this domain organization and the antiproliferative activity was investigated by generating a hybrid protein with the 204 a domain upstream from the 203 b domain. This 204a/203b protein inhibits the proliferation of transfected cells, delays G0/G1 progression into S phase following serum restimulation, and inhibits the E2F-mediated transcriptional activity. These results demonstrate for the first time that both a and b domains are needed for inhibition of proliferation by the Ifi 200 proteins. PMID- 10452525 TI - Localization of acidic residues involved in the proton pumping activity of the bovine heart mitochondrial bc1 complex. AB - Chemical modification of carboxyl residues in polypeptide subunits of the mitochondrial bc1 complex causes a decoupling effect, that is inhibition of the proton pumping activity, without affecting the rate of electron transfer to ferricytochrome c. The study presented here is aimed at localizing and identifying the residues whose modification results in decoupling of the complex. Glutamate-53 in subunit IX (the DCCD-binding protein) and aspartate-166 in the Rieske iron-sulfur protein are the residues modified by N,N' dicyclohexylcarbodiimide (DCCD) and N-(ethoxycarbonyl)-2-ethoxy-1,2 dihydroquinoline (EEDQ), respectively. The results obtained also suggest that the carboxy-terminal sequence of the Core protein II, which is fairly rich in acidic residues, may also play a role in the vectorial proton translocation activity of the complex. PMID- 10452526 TI - Short-term block of Na+/K+-ATPase in neuro-glial cell cultures of cerebellum induces glutamate dependent damage of granule cells. AB - Granule cells in a dissociated neuro-glial cell culture of cerebellum when exposed to ouabain (10(-3) M) for 25 min apparently swell, increase their [Ca2+]i with obvious depolarization of the mitochondrial membrane. In 3 h after ouabain was omitted from the solution, 62 +/- 3% of granule cells had pycnotic nuclei. The supplement of a solution with competitive specific antagonist of NMDA receptors, L-2-amino-7-phosphonoheptanoate (10(-4) M, APH) together with ouabain prevented cells from swelling, mitochondrial deenergization, neuronal death and increase of [Ca2+]i. These data suggest that cellular Na+/K+-ATPase inactivation in neuro-glial cell cultures of cerebellum leads to glutamate (Glu) accumulation, hyperstimulation of glutamate receptors, higher Ca2+ and Na+ influxes into the cells through the channels activated by Glu. This process leads to cell swelling, mitochondrial deenergization and death of granule cells. Possibly, the decrease of Na+/K+-ATPase activity in brain cells can lead to the onset of at least some chronic neurological disorders. PMID- 10452527 TI - PKC-beta and PKC-zeta mediate opposing effects on proximal tubule Na+,K+-ATPase activity. AB - Dopamine (DA) inhibits rodent proximal tubule Na+,K+-ATPase via stimulation of protein kinase C (PKC). However, direct stimulation of PKC by phorbol 12 myristate 13-acetate (PMA) results in increased Na+,K+-ATPase. LY333531, a specific inhibitor of the PKC-beta isoform, prevents PMA-dependent activation of Na+,K+-ATPase, but has no effect on DA inhibition of this activity. A similar result was obtained with a PKC-beta inhibitor peptide. Concentrations of staurosporine, that inhibits PKC-zeta, prevent DA-dependent inhibition of Na+,K+ ATPase and a similar effect was obtained with a PKC-zeta inhibitor peptide. Thus, PMA-dependent stimulation of Na+,K+-ATPase is mediated by activation of PKC-beta, whereas inhibition by DA requires activation of PKC-zeta. PMID- 10452528 TI - 2.0 A X-ray structure of the ternary complex of 7,8-dihydro-6 hydroxymethylpterinpyrophosphokinase from Escherichia coli with ATP and a substrate analogue. AB - The X-ray crystal structure of 7,8-dihydro-6-hydroxymethylpterinpyrophosphokinase (PPPK) in a ternary complex with ATP and a pterin analogue has been solved to 2.0 A resolution, giving, for the first time, detailed information of the PPPK/ATP intermolecular interactions and the accompanying conformational change. The first 100 residues of the 158 residue peptide contain a betaalpha betabeta alphabeta motif present in several other proteins including nucleoside diphosphate kinase. Comparative sequence examination of a wide range of prokaryotic and lower eukaryotic species confirms the conservation of the PPPK active site, indicating the value of this de novo folate biosynthesis pathway enzyme as a potential target for the development of novel broad-spectrum anti-infective agents. PMID- 10452529 TI - A missense mutation in canine C1C-1 causes recessive myotonia congenita in the dog. AB - Myotonia congenita is an inherited disorder of sarcolemmal excitation leading to delayed relaxation of skeletal muscle following contractions. Mutations in a skeletal muscle voltage-dependent chloride channel, CIC-1, have been identified as the molecular genetic basis for the syndrome in humans, and in two well characterized animal models of the disease: the myotonic goat, and the arrested development of righting (adr) mouse. We now report the molecular genetic and electrophysiological characterization of a canine CIC-1 mutation that causes autosomal recessive myotonia congenita in miniature Schnauzers. The mutation results in replacement of a threonine residue in the D5 transmembrane segment with methionine. Functional characterization of the mutation introduced into a recombinant CIC-1 and heterologously expressed in a cultured mammalian cell line demonstrates a profound effect on the voltage-dependence of activation such that mutant channels have a greatly reduced open probability at voltages near the resting membrane potential of skeletal muscle. The degree of this dysfunction is greatly diminished when heterodimeric channels containing a wild-type and mutant subunit are expressed together as a covalent concatemer strongly supporting the observed recessive inheritance in affected dog pedigrees. Genetic and electrophysiological characterization of the myotonic dog provides a new and potentially valuable animal model of an inherited skeletal muscle disease that has advantages over existing models of myotonia congenita. PMID- 10452530 TI - Oligonucleotide-peptide conjugates as potential antisense agents. AB - Oligonucleotide-peptide conjugates have several applications, including their potential use as improved antisense agents for interfering with the RNA function within cells. In order to provide robust and generally applicable conjugation chemistry, we developed a novel approach of fragment coupling of pre-synthesized peptides to the 2'-position of a selected nucleotide within an otherwise protected oligonucleotide chain attached to a solid support. PMID- 10452531 TI - Serotonin 5-HT1B and 5-HT1D receptors form homodimers when expressed alone and heterodimers when co-expressed. AB - The serotonin (5-hydroxytryptamine (5-HT)) 1B and 1D receptor subtypes share a high amino acid sequence identity and have similar ligand binding properties. In this study, we demonstrate that both receptor subtypes exist as monomers and homodimers when expressed alone and as monomers and heterodimers when co expressed. Gene expression studies have shown that there are brain regions where the 5-HT1B and 5-HT1D receptors are co-localized and where heterodimerization may occur physiologically. This is the first direct visualization of the physical association between G protein-coupled receptors of different subtypes. PMID- 10452532 TI - Subunit C of the vacuolar H+-ATPase of Hordeum vulgare. AB - The molecular cloning of the first subunit C of the plant vacuolar H+-ATPase is reported. Tonoplast vesicles were purified from barley leaves by sucrose gradient centrifugation, and the tonoplast polypeptides were separated by two-dimensional (2-D) gel electrophoresis. Using an anti-ATPase holoenzyme antibody, a polypeptide was recognized in the molecular mass range of 40 kDa with an isoelectric point of about 6.0, and tentatively identified as subunit C. The polypeptide spot was excised from about 50 2-D gels and subjected to endo Lys C proteolysis. Two proteolytic peptides were sequenced and the amino acid sequences were used to design degenerated oligonucleotides, followed by PCR amplification with cDNA template and screening of a cDNA library synthesized from Hordeum vulgare poly A mRNA of epidermis strips. The full length clone of 1.5 kbp contains an open reading frame of 1062 bp encoding a polypeptide of 354 amino acids with a molecular mass of 39,982 Da and an isoelectric point of 6.04. Amino acid identity with sequences of SUC from animals and fungi is in the range of 36.7 to 38.5%. Expression of the cloned gene was demonstrated by Northern blotting and RT-PCR. PMID- 10452533 TI - The low Mr phosphotyrosine protein phosphatase behaves differently when phosphorylated at Tyr131 or Tyr132 by Src kinase. AB - The low molecular weight phosphotyrosine protein phosphatase (LMW-PTP) is phosphorylated by Src and Src-related kinases both in vitro and in vivo; in Jurkat cells, and in NIH-3T3 cells, it becomes tyrosine-phosphorylated upon stimulation by PDGF. In this study we show that pp60Src phosphorylates in vitro the enzyme at two tyrosine residues, Tyr131 and Tyr132, previously indicated as the main phosphorylation sites of the enzyme, whereas phosphorylation by the PDGF R kinase is much less effective and not specific. The effects of LMW-PTP phosphorylation at each tyrosine residue were investigated by using Tyr131 and Tyr132 mutants. We found that the phosphorylation at either residue has differing effects on the enzyme behaviour: Tyr131 phosphorylation is followed by a strong (about 25-fold) increase of the enzyme specific activity, whereas phosphorylation at Tyr132 leads to Grb2 recruitment. These differing effects are discussed on the light of the enzyme structure. PMID- 10452534 TI - Mapping of putative binding sites on the ectodomain of the type II TGF-beta receptor by scanning-deletion mutagenesis and knowledge-based modeling. AB - Binding surfaces of the type II transforming growth factor (TGF)-beta receptor extracellular domain (TbetaRII-ECD) are mapped by combining scanning-deletion mutagenesis results with knowledge-based modeling of the ectodomain structure. Of the 17 deletion mutants produced within the core binding domain of TbetaRII-ECD, only three retained binding to TGF-beta. Comparative modeling based on the crystal structure of the activin type II receptor extracellular domain (ActRII ECD) indicates that the TbetaRII mutants which retain TGF-beta binding are deleted in some of the loops connecting the beta-strands in the TbetaRII-ECD model. Interpretation of the mutagenesis data within the structural framework of the ectodomain model allows for the prediction of potential binding sites at the surface of TbetaRII-ECD. PMID- 10452535 TI - Motifer, a search tool for finding amino acid sequence patterns from nucleotide sequence databases. AB - Motifer is a software tool able to find directly in nucleotide databases very distant homologues to an amino acid query sequence. It focuses searches on a specific amino acid pattern, scoring the matching and intervening residues as specified by the user. The program has been developed for searching databases of expressed sequence tags (ESTs), but it is also well suited to search genomic sequences. The query sequence can be a variable pattern with alternative amino acids or gaps and the sequences searched can contain introns or sequencing errors with accompanying frame shifts. Other features include options to generate a searchable output, set the maximal sequencing error frequency, limit searches to given species, or exclude already known matches. Motifer can find sequence homologues that other search algorithms would deem unrelated or would not find because of sequencing errors or a too large number of other homologues. The ability of Motifer to find relatives to a given sequence is exemplified by searches for members of the transforming growth factor-beta family and for proteins containing a WW-domain. The functions aimed at enhancing EST searches are illustrated by the 'in silico' cloning of a novel cytochrome P450 enzyme. PMID- 10452536 TI - Enzymatic synthesis of beta-lactam antibiotics using penicillin-G acylase in frozen media. AB - Penicillin-G acylase (EC 3.5.1.11) from Escherichia coli catalyzed the synthesis of various beta-lactam antibiotics in ice at -20 degrees C with higher yields than obtained in solution at 20 degrees C. The initial ratio between aminolysis and hydrolysis of the acyl-enzyme complex in the synthesis of cephalexin increased from 1.3 at 20 degrees C to 25 at -20 degrees C. The effect on the other antibiotics studied was less, leading us to conclude that freezing of the reaction medium influences the hydrolysis of each nucleophile-acyl-enzyme complex to a different extent. Only free penicillin-G acylase could perform transformations in frozen media: immobilized preparations showed a low, predominantly hydrolytic activity under these conditions. PMID- 10452537 TI - The sensitivity of versican from rabbit lung to gelatinase A (MMP-2) and B (MMP 9) and its involvement in the development of hydraulic lung edema. AB - Large chondroitinsulphate-containing proteoglycan (versican) isolated from rabbit lung was cleaved by purified gelatinase A (MMP-2) and gelatinase B (MMP-9), as well as by crude enzyme extract from rabbit lung with hydraulic edema. Gelatine zymography, performed after purification of gelatinases by affinity chromatography, demonstrated that the enzyme extract contained two main gelatinolytic bands at about 92 kDa and 72 kDa, identified by specific antisera as the latent proMMP-9 and proMMP-2, respectively. Moreover, enzyme extract from edematous lung showed an increased amount of the proteolytically activated forms of both gelatinases with respect to normal controls. These results suggest that MMP-2 and MMP-9 are involved in the breakdown of versican occurring in rabbit lung during the development of hydraulic edema. PMID- 10452538 TI - Occurrence of an HIV-1 gp160 endoproteolytic activity in low-density vesicles and evidence for a distinct density distribution from endogenously expressed furin and PC7/LPC convertases. AB - Human immunodeficiency virus (HIV) glycoprotein (gp) 160 processing by host cell proteinases is an essential step for viral fusion and infectivity. We have identified a rat liver subcellular fraction which specifically processes gp160 into gp120 and gp41. Using equilibration of microsomes in sucrose gradients, the gp160 cleavage activity was associated with particles equilibrating at low densities, well-separated from the endoplasmic reticulum, cis-Golgi network, Golgi stacks, lysosomes and plasma membrane. Its density distribution was compatible with light secretory vesicles derived from the trans-Golgi network (TGN) or to endosomes, but association with endosomes was not supported by free flow electrophoresis. Although furin and pro-protein convertase (PC) 7/LPC have been proposed as the major gp160 processing convertases, the rat liver microsomal gp160 processing activity was essentially resolved from furin and only partially overlapped PC7/LPC. These data suggest that proteinase(s) other than furin and PC7/LPC, presumably located in TGN-derived vesicles, may participate in the gp160 processing into gp120 and gp41. PMID- 10452540 TI - The X gene of hepatitis B virus shows a high level stimulation of the Rous sarcoma virus long terminal repeat in the methylotropic yeast, Pichia pastoris. AB - In order to study the transactivational property of the X gene in the methylotropic yeast Pichia pastoris, a Rous sarcoma virus-chloramphenicol acetyltransferase (RSV-CAT) cassette was co-transformed and integrated into the host yeast strain as a reporter which showed an overwhelming CAT activity. Immunoprecipitation of the yeast cell extracts with an X-specific monoclonal antibody, however, showed a low level expression of the X gene. Therefore besides a trans-effect of the X protein, the enhanced reporter activity could be a manifestation of a cis-effect of the X gene sequences also. Therefore, unlike the transactivation studies with X gene in animal cells where limited functional activity is observed, P. pastoris appears to be an excellent system to study cis- and trans-aspects of gene regulation by the X gene. PMID- 10452539 TI - Endothelin-1 induces expression of fetal genes through the interleukin-6 family of cytokines in cardiac myocytes. AB - We here examined the role of the interleukin-6 (IL-6) family of cytokines in endothelin-1 (ET-1)-induced hypertrophic responses using cultured cardiac myocytes of neonatal rats. ET-1 induced expression of IL-6 and leukemia inhibitory factor (LIF) genes. ET-1-induced LIF gene expression was abolished by inhibition of protein kinase C activity. ET-1 activated the promoter of atrial natriuretic peptide and beta-type myosin heavy chain genes through the tyrosine kinase pathway and IL-6 receptor gp130. These results suggest that the IL-6 family of cytokines mediates ET-1-induced expression of some fetal genes in cardiac myocytes. PMID- 10452541 TI - Oxythiamine and dehydroepiandrosterone induce a G1 phase cycle arrest in Ehrlich's tumor cells through inhibition of the pentose cycle. AB - Transketolase (TK) reactions play a crucial role in tumor cell nucleic acid ribose synthesis utilizing glucose carbons, yet, current cancer treatments do not target this central pathway. Experimentally, a dramatic decrease in tumor cell proliferation after the administration of the TK inhibitor oxythiamine (OT) was observed in several in vitro and in vivo tumor models. Here, we demonstrate that pentose cycle (PC) inhibitors, OT and dehydroepiandrosterone (DHEA), efficiently regulate the cell cycle and tumor proliferation processes. Increasing doses of OT or DHEA were administered by daily intraperitoneal injections to Ehrlich's ascites tumor hosting mice for 4 days. The tumor cell number and their cycle phase distribution profile were determined by DNA flow histograms. Tumors showed a dose dependent increase in their G0-G1 cell populations after both OT and DHEA treatment and a simultaneous decrease in cells advancing to the S and G2-M cell cycle phases. This effect of PC inhibitors was significant, OT was more effective than DHEA, both drugs acted synergistically in combination and no signs of direct cell or host toxicity were observed. Direct inhibition of PC reactions causes a G1 cell cycle arrest similar to that of 2-deoxyglucose treatment. However, no interference with cell energy production and cell toxicity is observed. PC inhibitors, specifically ones targeting TK, introduce a new target site for the development of future cancer therapies to inhibit glucose utilizing pathways selectively for nucleic acid production. PMID- 10452542 TI - The evolution of starch-binding domain. AB - Amylolytic enzymes belonging to three distinct families of glycosidases (13, 14, 15) contain the starch-binding domain (SBD) positioned almost exclusively at the C-terminus. Detailed analysis of all available SBD sequences from 43 different amylases revealed its independent evolutionary behaviour with regard to the catalytic domains. In the evolutionary tree based on sequence alignment of the SBDs, taxonomy is respected so that fungi and actinomycetes form their own separate parts surrounded by bacteria that are also clustered according to taxonomy. The only known N-terminal SBD from Rhizopus oryzae glucoamylase is on the longest branch separated from all C-terminal SBDs. The 3-dimensional (3-D) structures of fungal glucoamylase and bacterial CGTase SBDs are compared and used to discuss the interesting SBD evolution. PMID- 10452543 TI - Monoclonal antibody to the delta opioid receptor acts as an agonist in dual regulation of adenylate cyclase in NG108-15 cells. AB - Monoclonal antibodies generated against multiple antigenic peptides of the N terminal sequence (3LVPSARAELQSSPLV17) of the cloned delta opioid receptor immunoprecipitated a 58 kDa protein from CHAPS-solubilized NG108-15 membranes. The immunoprecipitates bound [3H]DPDPE--but not [3H]DAMGO--with a Kd of 6.4 nM and a Bmax of 75 pM. Western blot analysis revealed a distinct band of 58 kDa. The antibodies inhibited basal and PGE1-stimulated cAMP levels, and mimicked the effect of agonists manifest in a compensatory increase in cAMP formation. The antibody will be potentially useful in the analysis of functional epitopes on the delta opioid receptor. PMID- 10452544 TI - Transactivation of the WT1 antisense promoter is unique to the WT1[+/-] isoform. AB - The Wilms' tumour suppressor gene, WT1, encodes a zinc finger transcription factor that has been shown to repress a variety of cellular promoters via binding to cognate DNA elements. Our earlier work identified an antisense WT1 promoter that contains WT1 consensus sites, but is transcriptionally activated by WT1. In this study, we demonstrate that, unlike previous reports of transcriptional regulation by WT1, transactivation of the antisense promoter is unique to a single isoform of WT1. Of the four alternatively spliced isoforms in which exon 5 (at splice I) or amino acid residues KTS (at splice II) are inserted or omitted, only the WT1 isoform containing splice I and omitting splice II (WT1[+/-]) displays transactivation. We demonstrate that transregulation variations observed with WT1 isoforms are not solely attributable to differential DNA binding by [+KTS] or [-KTS] isoforms. Thus, the transactivation of the antisense promoter displays an absolute requirement for exon 5, suggesting that interaction between WT1 and other cellular factors is necessary for this regulatory function. PMID- 10452545 TI - Persistence of botulinum neurotoxin action in cultured spinal cord cells. AB - Primary dissociated fetal mouse spinal cord cultures were used to study the mechanisms underlying the differences in persistence of botulinum neurotoxin A (BoNT/A) and botulinum neurotoxin/E (BoNT/E) activities. Spinal cord cultures were exposed to BoNT/A (0.4 pM) for 2-3 days, which converted approximately half of the SNAP-25 to an altered form lacking the final nine C-terminal residues. The distribution of toxin-damaged to control SNAP-25 remained relatively unchanged for up to 80 days thereafter. Application of a high concentration of BoNT/E (250 pM) either 25 or 60 days following initial intoxication with BoNT/A converted both normal and BoNT/A-truncated SNAP-25 into a single population lacking the final 26 C-terminal residues. Excess BoNT/E was removed by washout, and recovery of intact SNAP-25 was monitored by Western blot analysis. The BoNT/E-truncated species gradually diminished during the ensuing 18 days, accompanied by the reappearance of both normal and BoNT/A-truncated SNAP-25. Return of BoNT/A truncated SNAP-25 was observed in spite of the absence of BoNT/A in the culture medium during all but the first 3 days of exposure. These results indicate that proteolytic activity associated with the BoNT/A light chain persists inside cells for > 11 weeks, while recovery from BoNT/E is complete in < 3 weeks. This longer duration of enzymatic activity appears to account for the persistence of serotype A action. PMID- 10452546 TI - Effect of protein disulfide isomerase on the regeneration of bovine ribonuclease A with dithiothreitol. AB - The role of protein disulfide isomerase (PDI) in the regeneration of ribonuclease A with dithiothreitol (DTT) was investigated at three different temperatures. The rates of formation of the native protein were markedly increased in the presence of PDI, 9-fold at 15 degrees C, 6-fold at 25 degrees C and 62-fold at 37 degrees C, respectively. In the presence of PDI, major changes were found in the distribution of intermediates in the three-disulfide region at 25 and 15 degrees C and also in the one-disulfide region at 15 degrees C, with the fast accumulation of the two native-like species des-[65-72] and des-[40-95]. The present results indicate that PDI does not alter the two major parallel pathways involving des-[65-72] and des-[40-95] in the regeneration of ribonuclease A with DTT. PMID- 10452547 TI - A novel K+ channel blocking toxin from Tityus discrepans scorpion venom. AB - A novel toxin (TdK1) was purified from the venom of the scorpion Tityus discrepans, sequenced and functionally characterized. It contains 37 amino acid residues and blocks reversible the shakerB K+ channel expressed in SF9 cells with a Kd in the order of 280 nM. The proposed systematic nomenclature for this peptide is alpha-KTx4.3. PMID- 10452548 TI - Involvement of Sp1 in basal and retinoic acid induced transcription of the human tissue-type plasminogen activator gene. AB - Transcription of the human tissue-type plasminogen activator (t-PA) gene is regulated by a multi-hormonal responsive enhancer at -7 kb. Transient co transfections of Drosophila SL2 and human HT1080 fibrosarcoma cells with t-PA reporter constructs showed that Sp1 and Sp3 activate the t-PA promoter. Moreover Sp1 (but not Sp3) binding to the promoter is involved in induction by retinoic acid (RA), a response mediated through the enhancer. The role of Sp1 is specific, since mutation of the CRE element in the promoter did not affect response to RA. In contrast, the glucocorticoid induction mediated by the enhancer is independent of these Sp1 and CRE elements. PMID- 10452550 TI - Co-introduction of an antisense gene for an endogenous seed storage protein can increase expression of a transgene in Arabidopsis thaliana seeds. AB - We have investigated whether the expression in Arabidopsis thaliana seeds of a transgene (the Phaseolus vulgaris arcelin (arc)5-I gene) could be enhanced by the simultaneous introduction of an antisense gene for an endogenous seed storage protein (2S albumin). Seeds of plants transformed with both the arc5-I gene and a 2S albumin antisense gene contained reduced amounts of 2S albumins and increased arcelin-5 (Arc5) accumulation levels compared to lines harboring the arc5-I gene only. Arc5 production could be enhanced to more than 24% of the total seed protein content, suggesting that antisense technology could be of great utility to favor high expression of transgenes. PMID- 10452549 TI - The trefoil peptides TFF2 and TFF3 are expressed in rat lymphoid tissues and participate in the immune response. AB - Members of the trefoil factor (TFF) family are mucin-associated polypeptides that are expressed along the entire length of the gastrointestinal tract. TFFs have been proposed to play a role in mucosal defence through both protective and reparative mechanisms. The potential relationship between TFFs and mucins in non gut glycoprotein-secreting epithelia has not been fully explored. In the present study we identified TFF2 and TFF3 mRNA and peptide in rat lymphoid tissues, demonstrated that TFF peptide expression in rat spleen increased 1.5- to 3-fold following experimental induction of the immune response, and showed that hTFF2 and hTFF3 (1-5 mg/ml) stimulated migration of human monocytes. Our data suggest that TFFs may in part be involved in the repair of injury through the modulation of the inflammatory response. PMID- 10452551 TI - Generation of multiple transcripts from the chicken chondromodulin-I gene and their expression during embryonic development. AB - Chondromodulin-I (ChM-I) is an angiogenesis inhibitor isolated from fetal bovine cartilage. Here, we report the nucleotide sequence of chicken ChM-I cDNA. Chicken mature ChM-I had a significantly larger N-terminal hydrophilic domain than its mammalian counterparts. Chicken embryos expressed multiple transcripts (3.3, 2.0 and 1.7 kb in size) due to the alternative utilization of polyadenylation signals, whereas only the 1.7 kb transcripts were detected in mammals. Although confined to cartilage and eye at a later stage of development, whole-mount in situ hybridization revealed the expression of ChM-I mRNA in somites, heart, bronchial arches, roof plate, retina and limb buds. The expression pattern of the gene suggests a role for ChM-I in the morphogenesis during embryonic development. PMID- 10452552 TI - Organisation of mitochondria in living sensory neurons. AB - In this work, we have examined the mitochondrial organisation in living cultured primary dorsal root ganglion (DRG) neurons. Confocal microscopy and the mitochondrial potential-sensitive fluorescent dye 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazolo carbocyanine iodide (JC-1) were used to visualise intracellular structures with a high and low membrane potential. Three dimensional reconstruction revealed a mitochondrial organisation featuring separate highly polarised mitochondria, clusters of mitochondria located mainly at the base of neurite hillocks and filamentous mitochondrial structures. Filamentous mitochondria were distributed along the cell body, especially between neurites. A functional integration between mitochondrial structures is proposed. PMID- 10452553 TI - Ap4A induces apoptosis in human cultured cells. AB - Diadenosine oligophosphates (Ap(n)A) have been proposed as intracellular and extracellular signaling molecules in animal cells. The ratio of diadenosine 5',5'''-P1,P3-triphosphate to diadenosine 5',5'''-P1,P4-tetraphosphate (Ap3A/Ap4A) is sensitive to the cellular status and alters when cultured cells undergo differentiation or are treated with interferons. In cells undergoing apoptosis induced by DNA topoisomerase II inhibitor VP16, the concentration of Ap3A decreases significantly while that of Ap4A increases. Here, we have examined the effects of exogenously added Ap3A and Ap4A on apoptosis and morphological differentiation. Penetration of Ap(n)A into cells was achieved by cold shock. Ap4A at 10 microM induced programmed cell death in human HL60, U937 and Jurkat cells and mouse VMRO cells and this effect appeared to require Ap4A breakdown as hydrolysis-resistant analogues of Ap4A were inactive. On its own, Ap3A induced neither apoptosis nor cell differentiation but did display strong synergism with the protein kinase C activators 12-deoxyphorbol-13-O-phenylacetate and 12 deoxyphorbol-13-O-phenylacetate-20-acetate in inducing differentiation of HL60 cells. We propose that Ap4A and Ap3A are physiological antagonists in determination of the cellular status: Ap4A induces apoptosis whereas Ap3A is a co inductor of differentiation. In both cases, the mechanism of signal transduction remains unknown. PMID- 10452554 TI - Peptide models for inherited neurodegenerative disorders: conformation and aggregation properties of long polyglutamine peptides with and without interruptions. AB - Several neurodegenerative diseases are caused by expansion of polyglutamine repeats in the affected proteins. In spino-cerebellar ataxia type 1 (SCA1), histidine interruptions have been reported to mitigate the pathological effects of long glutamine stretches. To understand this phenomenon, we investigated the conformational preferences of peptides containing both the uninterrupted polyglutamine stretches and those with histidine interruption(s) as seen in SCA1 normals. Our study suggests that substitution of histidines by glutamines induces a conformational change which results in decreased solubility and increased aggregation. Our findings also suggest that all the polyglutamine peptides with and without interruption(s) adopt a beta-structure and not random coil. PMID- 10452555 TI - Inhibition of HIV-1 replication by a two-strand system (FTFOs) targeted to the polypurine tract. AB - Reverse transcription of HIV-1 vRNA into the double-stranded DNA provirus involves initiation of plus-strand DNA synthesis at the polypurine tract (PPT) by reverse transcriptase (RT). The PPT is highly conserved among the known human immunodeficiency virus (HIV-1) strains and is a possible target for triplex formation. We show the effects of triple-helix formation by assays of primer extension inhibition in vitro, using a two-strand system (foldback triplex forming oligonucleotides (FTFOs)) targeted to the PPT of HIV-1. The two-stranded composition of a triple-helix is thermodynamically and kinetically superior to the three-strand system. The FTFOs inhibited the RT activity in a sequence specific manner, i.e. the triplex actually formed at the PPT and blocked the RT. The FTFOs containing the phosphorothioate groups at the antisense sequences showed greater 3'-exonuclease resistance. In HIV-1-infected MOLT-4 cells, the FTFOs containing the phosphorothioate groups at the antisense sequence sites and guanosine rich parts within the third Hoogsteen base-pairing sequence inhibit the replication of HIV-1 more effectively than the antisense oligonucleotides, indicating sequence-specific inhibition of HIV-1 replication. PMID- 10452556 TI - Opening the Schiff base moiety of bacteriorhodopsin by mutation of the four extracellular Glu side chains. AB - The quadruple bacteriorhodopsin (BR) mutant E9Q+E74Q+E194Q+E204Q shows a lambda(max) of about 500 nm in water at neutral pH and a great influence of pH and salts on the visible absorption spectrum. Accessibility to the Schiff base is strongly increased, as detected by the rapid bleaching effect of hydroxylamine in the dark as well as in light. Both the proton release kinetics and the photocycle are altered, as indicated by a delayed proton release after proton uptake and changed M kinetics. Moreover, affinity of the color-controlling cation(s) is found to be decreased. We suggest that the four Glu side chains are essential elements of the extracellular structure of BR. PMID- 10452557 TI - MAP kinase activation by mu opioid receptor involves phosphatidylinositol 3 kinase but not the cAMP/PKA pathway. AB - The involvement of protein kinases was studied in mu opioid receptor activation of mitogen-activated protein (MAP) kinase using cells transfected with the receptor clone. The cAMP/protein kinase A (PKA) pathway is known to be the major biochemical pathway for mu opioid receptor signaling. However, our data showed that stimulating adenylyl cyclase or activating PKA had no effect on mu receptor enhancement of MAP kinase activity, suggesting that the cAMP/PKA pathway is not involved in mediating the mu receptor activation of MAP kinase. Inhibition of phosphatidylinositol (PI) 3-kinase reduced mu receptor enhancement of MAP kinase activity, suggesting PI 3-kinase involvement. Together, these results show that cross-talk between the mu opioid receptor and the MAP kinase cascade is not mediated by the cAMP/PKA pathway, but involves PI 3-kinase. PMID- 10452558 TI - Activation of src tyrosine kinases by peroxynitrite. AB - In this study, we demonstrate that the phosphorylation activity of five tyrosine kinases of the src family from both human erythrocytes (lyn, hck and c-fgr) and bovine synaptosomes (lyn and fyn) was stimulated by treatment with 30-250 microM peroxynitrite. This effect was not observed with syk, a non-src family tyrosine kinase. Treatment of kinase immunoprecipitates with 0.01-10 microM peroxynitrite showed that the interaction of these enzymes with the oxidant also activated the src kinases. Higher concentrations of peroxynitrite inhibited the activity of all kinases, indicating enzyme inactivation. The addition of bicarbonate (1.3 mM CO2) did not modify the upregulation of src kinases but significantly protected the kinases against peroxynitrite-mediated inhibition. Upregulation of src kinase activity by 1 microM peroxynitrite was 3.5-5-fold in erythrocytes and 1.2-2-fold in synaptosomes, but this could be the result, at least in part, of the higher basal level of src kinase activity in synaptosomes. Our results indicate that peroxynitrite can upregulate the tyrosine phosphorylation signal through the activation of src kinases. PMID- 10452559 TI - Evidence for the presence of several phosphodiesterase isoforms in brown adipose tissue of Zucker rats: modulation of PDE2 by the fa gene expression. AB - The present study was undertaken to characterise the phosphodiesterases (PDEs) present in brown adipose tissue (BAT) of Zucker rat pups and to determine whether the capacity for degradation of cyclic nucleotides was affected by the fatty genotype. Regardless of the genotype, PDE2-4 contributed to total PDE activity, the PDE3 activity equalling the sum of PDE2 and 4 activities. In fa/fa compared to Fa/fa rats, (a) PDE2 activity was significantly increased, (b) Western blot analysis of PDE2 revealed two signals at 71 and 105 kDa, with changes in protein being in good parallelism with changes in activity, (c) the PDE2 mRNA concentration was also significantly increased. In good agreement, the cGMP concentration was decreased in BAT from fa/fa pups. PMID- 10452560 TI - ATP-dependent degradation of SulA, a cell division inhibitor, by the HslVU protease in Escherichia coli. AB - HslVU is an ATP-dependent protease consisting of two multimeric components, the HslU ATPase and the HslV peptidase. To gain an insight into the role of HslVU in regulation of cell division, the reconstituted enzyme was incubated with SulA, an inhibitor of cell division in Escherichia coli, or its fusion protein with maltose binding protein (MBP). HslVU degraded both proteins upon incubation with ATP but not with its nonhydrolyzable analog, ATPgammaS, indicating that the degradation of SulA requires ATP hydrolysis. The pulse-chase experiment using an antibody raised against MBP-SulA revealed that the stability of SulA increased in hsl mutants and further increased in lon/hsl double mutants, indicating that SulA is an in vivo substrate of HslVU as well as of protease La (Lon). These results suggest that HslVU in addition to Lon plays an important role in regulation of cell division through degradation of SulA. PMID- 10452561 TI - Propeptide of the metalloprotease of Brevibacillus brevis 7882 is a strong inhibitor of the mature enzyme. AB - A metalloprotease gene of Brevibacillus brevis (npr) was expressed in Escherichia coli in a soluble form as native Npr precursor. A significant fraction of the precursor was spontaneously processed, producing the N-terminal propeptide and the mature enzyme. A strong inhibition of the mature Npr by its own propeptide in the crude lysate was observed even in the absence of the covalent linkage between them. Pure precursor, propeptide and the mature Npr were isolated and kinetic parameters of the mature enzyme inhibition by the propeptide were determined. The inhibition is of the tight-binding competitive type with Ki 0.17 nM. Inhibition of metalloproteases from Brevibacillus megaterium and thermolysine by the heterologous propeptide of the Npr from B. brevis was much weaker or none. PMID- 10452562 TI - Introduction to symposium on chronic obstructive pulmonary disease. PMID- 10452563 TI - The pathogenesis of chronic obstructive pulmonary disease. AB - Cigarette smoking is the main risk factor for the development of chronic obstructive pulmonary disease (COPD). An accelerated rate of lung function decline that causes clinically significant COPD, however, is present in only a minority of smokers. In addition to the cumulative amount of cigarettes smoked, other environmental and genetic properties contribute to this variable physiological response. This article reviews the role of airway hyperresponsiveness, mucus hypersecretion, infection, and proteases in the development of COPD. PMID- 10452564 TI - Outpatient management of chronic obstructive pulmonary disease. AB - The outpatient management of chronic obstructive pulmonary disease (COPD) is designed to limit the decline in respiratory function over time, to relieve the symptoms and improve the patient's functional status, and to manage complications when they arise. Factors that predispose to airway inflammation, including cigarette smoking and respiratory infections, are prevented by behavioral modification programs, measures such as exercise and nutrition to improve general health, and regular vaccination. Symptoms are relieved by bronchodilator and anti inflammatory therapy, based upon the specific needs of the patient. Hypoxemia and acute infections are treated with oxygen administration and the use of antibiotics when necessary. The management of acute exacerbations of COPD is addressed elsewhere in this symposium (ie, choice of antibiotics is not discussed here). Also, certain aspects of management, such as surgical procedures, chest physical therapy, and other aspects of pulmonary rehabilitation, are also subjects of subsequent articles in this series. Although none of these modalities, except for smoking cessation and oxygen administration, have been shown to alter the course of COPD, the careful choice of the therapeutic measures discussed here can lead to significant relief of symptoms in the patient with chronic airway obstruction. PMID- 10452565 TI - The role of antimicrobial therapy in acute exacerbations of chronic bronchitis. AB - Acute exacerbations of chronic bronchitis (AECB) result in increased morbidity and mortality. The role of bacteria in AECB, the importance of antimicrobial therapy, and the choice of antimicrobial agents have been debated for decades. Fortunately, within the past few years, a number of studies and one consensus statement have been reported that have increased the understanding of the role of bacteria in AECB and suggest approaches in selecting antimicrobial therapy. This article will review these studies and present an empiric approach in treating AECB based upon the patient's presenting findings, related risk factors, and potential antimicrobial resistance patterns that may be encountered. PMID- 10452566 TI - Surgical treatment of chronic obstructive pulmonary disease. AB - Over the past several decades, a number of surgical techniques have been developed for the treatment of chronic obstructive pulmonary disease. Many of these procedures have been abandoned because of lack of efficacy and/or high morbidity and mortality. At the present time, lung transplantation, reduction pneumoplasty for giant bullous emphysema, and lung volume reduction surgery are being performed in a number of centers. Data concerning the effectiveness of these procedures is accumulating and will ultimately need careful analysis to determine long-term outcomes in this group of patients. PMID- 10452567 TI - Alternative therapies for chronic bronchitis. AB - Alternative therapies for chronic bronchitis are therapies in which clinical efficacy is not well established. These can be broadly divided into expectorants, mucolytics, mucokinetics, antiproteases, antioxidants, and immunostimulants. Data supporting these therapies may be present in disorders clinically similar to chronic bronchitis, such as cystic fibrosis. Alternatively, these therapies may be based on clinical observations or in vitro studies that suggest a potential therapeutic benefit. Establishment of these therapies is likely to require more extensive investigation before their use is routinely recommended. Until such data is available, the clinician can only use his or her clinical judgement regarding the likely risk-benefit ratio. PMID- 10452568 TI - Pulmonary rehabilitation for the patient with severe chronic obstructive pulmonary disease. AB - Pulmonary rehabilitation measures acquired a poor reputation when early studies failed to demonstrate improvements in traditional pulmonary function tests. However, current evidence now indicates that such measures benefit patients with severe chronic obstructive pulmonary disease (COPD). Alleviation of symptoms, improved quality of life, improved nutrition, and decreased health care costs are some of the benefits seen in patients who successfully complete a rehabilitation program. This article reviews the role of a comprehensive rehabilitation program in the continuing care of the patient with severe COPD, the components of such a program, and specific measures to be considered. PMID- 10452569 TI - Course and prognosis of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) continues to increase in incidence as the population of developed countries ages. Much time has been devoted to studying the natural course of this disorder and to searching for risk factors for a decline in respiratory function. The major risk factor for a decline in 1 second forced expiratory volume (FEV1) is cigarette smoking, and smoking cessation is a major part of its prevention. Other risk factors for development of COPD include passive exposure to cigarette smoke, age, and genetic factors including airway hyperreactivity, eosinophilia, and a history of atopy. This discussion presents a review of the current literature regarding the natural course and prognosis of COPD. The approach to patients with end-stage disease and the use of advance directives is considered. PMID- 10452570 TI - Use of glycoprotein IIb/IIIa inhibitors in patients with coronary artery disease. AB - GP IIb/IIIa inhibitors are one of the most exciting pharmacological advances in the treatment of coronary artery disease to be developed this decade. They have proven benefit in the treatment of patients undergoing high-risk percutaneous interventions and in patients with unstable angina or non-Q-wave myocardial infarction. In all of these patients, they should be used in conjunction with aspirin and heparin. Although bleeding complications are high in initial studies, they are much less common with increased clinical experience and careful attention to heparin dosing. Important questions that will be answered in the next several years include whether there are significant differences between the available agents and whether oral formulations will be safe and effective. PMID- 10452571 TI - Changes in platelet ATP secretion and aggregation during pregnancy and in preeclampsia. AB - BACKGROUND: Platelet secretion plays an important role in the aggregation of platelets. However, the quantitative relationship between platelet aggregation and secretion of ATP during pregnancy and in pre-eclampsia has yet to be clarified. This study is designed to determine whether platelet count, volume, aggregation, and the amount of secreted ATP change in healthy, nonpregnant women, nonpreeclamptic pregnant women, and preeclamptic pregnant women and whether beta thromboglobulin (BTG) and platelet factor 4 (PF-4) concentrations alter in nonpreeclamptic and pre-eclamptic women. METHODS: Peripheral blood was collected from 114 women. Nonpreeclamptic pregnant women were divided into four groups (gestational weeks 10, 20, 30, and 35). Platelet aggregation and ATP secretion were investigated with the use of a lumi-aggregometer. BTG and PF-4 concentrations in peripheral blood were determined in 12 pregnant and 11 preeclamptic women. RESULTS: The amount of secreted ATP upon induction by 5 microM ADP increased significantly (P < 0.05-0.01) with gestational age. On the other hand, the amount of secreted ATP induced by 5 microg/mL collagen reached the maximal value from gestational weeks 20 to 35 in nonpreeclamptic women. Significantly more platelet aggregation was induced by the ADP and collagen in nonpreeclamptic women in gestational weeks 20 and 30 than in the gestational weeks 10 or 35 (P < 0.05-0.005). The amount of secreted ATP and platelet count were significantly lower (P < 0.05) in preeclampsia than in normal pregnancy. The BTG and PF-4 concentrations were significantly higher (P < 0.05) in preeclampsia than in normal pregnancy. CONCLUSIONS: The sensitivity of platelets for ATP secretion may intensify with progression of pregnancy. In normal pregnancy, around gestational week 35, the platelets may exhibit weaker ability to aggregate but maintain the capacity to secrete ATP. In preeclampsia, secreted ATP decreased because platelets may be stimulated to undergo a partial secretion. PMID- 10452572 TI - Acute transient chylous ascites associated with acute biliary pancreatitis. AB - We report a case of acute chylous ascites secondary to acute biliary pancreatitis, the first such case reported in the literature. Surprisingly, chylous ascites was detected during elective cholecystectomy. The pathogenesis and management of this problem is discussed. PMID- 10452573 TI - Neoplasms composed of eosinophilic polygonal cells: an overview with consideration of different cytomorphologic patterns. AB - Neoplasms that are composed of large eosinophilic cells present a distinctive and challenging image to the diagnostic pathologist. Potential lineages for such lesions include several types of epithelial tissue, as well as melanocytic, lymphoid, germ-cell, and mesenchymal proliferations. Attention to morphological details represents a useful first step in the further evaluation of eosinophilic large-cell tumors. These tumors can usually be segregated into one of four cytological categories, including granular eosinophilic (oncocytic and oncocytoid), fibrillary eosinophilic (myoid), globular eosinophilic (rhabdoid and gemistocytic), and diffusely eosinophilic (hyaline-cell and deeply eosinophilic, not further specified). Correlation with anatomic locations, nuances of clinical presentation, and results of electron microscopic and immunohistochemical studies provides additional data that can be used for a specific interpretation, especially if it is applied in an algorithmic fashion. PMID- 10452574 TI - Oncocytic and granular cell neoplasms of the central nervous system and pituitary gland. AB - Oncocytic transformation is an infrequent event within the central nervous system and is limited to neoplasms of the choroid plexus, meninges, and pituitary gland. Oncocytic modifications in choroid plexus tumors seem to occur predominantly in adult patients and in the fourth ventricle and do not seem to reflect any particular biological behavior. Meningiomas showing oncocytic differentiation have been recently described and this variant probably behaves more aggressively. Pituitary oncocytomas are regarded as a subtype of null cell adenomas. Oncocytic tumors have a significantly higher risk of progression with a higher recurrence rate after radiotherapy. Oncocytic changes in astrocytic neoplasms are rare. More frequently, astrocytomas can show granular changes that result in neoplasms composed of cells with granular cytoplasm and eccentric nuclei resembling foamy macrophages. Granular cell astrocytomas may mimic non-neoplastic lesions such as cerebral infarction and demyelinating disease. Cells with granular bodies are frequent in pilocytic astrocytoma, pleomorphic xanthoastrocytoma, and ganglion cell tumors. Their presence is considered a useful diagnostic finding to distinguish these low-grade lesions from malignant gliomas. A rare neoplasm is the granular cell tumor of the infundibulum that, when symptomatic, has to be differentiated from pituitary adenomas and other more common lesions of the sellar region. PMID- 10452575 TI - Oncocytic and oncocytoid tumors of the salivary glands. AB - Primary pink cell tumors of the salivary glands constitute a heterogeneous group of benign and malignant lesions characterized by tumor cells with abundant eosinophilic cytoplasm. These tumors are composed predominantly of oncocytic, epidermoid, or myoepithelial cells. Tumors with a significant oncocytic component include Warthin's tumor, oncocytoma, and oncocytic carcinoma. An epidermoid component can be seen as a metaplastic change or as a true cellular constituent of a mucoepidermoid carcinoma. Myoepithelial cells may have an epithelioid character and as a consequence may impart a pink cell appearance in pleomorphic adenoma, myoepithelioma, and myoepithelial carcinoma. Usually most of these tumors are fairly distinct morphologically and do not present diagnostic dilemmas. In select circumstances, especially when dealing with a limited tissue sample, a systematic approach with an appropriate immunohistochemical panel should be used in order to arrive at a correct diagnosis. Accurate assessment is the key in the subsequent management and follow-up of these patients. PMID- 10452576 TI - Oxyphilic proliferations of the respiratory tract and paranasal sinuses. AB - Neoplasms of the upper respiratory comprised primarily of eosinophilic cells are, in general, rare, and they include a diverse group of lesions. Low-grade oncocytic neuroendocrine neoplasms (so-called oncocytic carcinoids) can be encountered in several locations throughout the respiratory tract. The oncocytoma and related entities, lesions that presumably arise from minor gland tissue, can likewise be seen from the nasal cavity to the lung; differences in clinical significance may relate to the location of such lesions, and are discussed herein. Granular cell tumor is another entity that can involve both the upper respiratory tract and lungs, and specific features of this lesion in different anatomic sites are highlighted. The oncocytic variant of Schneiderian papilloma is an important nasal lesion to recognize, because of important therapeutic and prognostic implications of that diagnosis. Finally, unique oncocytic variants of glomus tumor and pulmonary alveolar adenoma are discussed, as well as eosinophilic varieties of pulmonary carcinomas and mesotheliomas. PMID- 10452577 TI - Eosinophilic and granular cell tumors of the breast. AB - Eosinophilic and granular cell tumors of the breast are a heterogeneous group encompassing both epithelial and mesenchymal lesions. A granular appearance of the cytoplasm may be caused by the accumulation of secretory granules, mitochondria, or lysosomes. In the breast, mucoid carcinomas, carcinomas showing apocrine differentiation, and neuroendocrine carcinomas are well known entities, while tumors with oncocytic and acinic cell differentiation have been only recently recognized. An abundance of lysosomes is characteristic of Schwannian granular cell neoplasms, but smooth muscle cell tumors also may have this cytoplasmic feature. Awareness of all these possibilities when granular cells are found in breast lesions improves diagnostic accuracy and helps to avoid misdiagnosis of both benign lesions and malignant tumors. PMID- 10452578 TI - Oncocytic and oncocytoid tumors of the exocrine pancreas, liver, and gastrointestinal tract. AB - Tumors having typical oncocytic features very rarely affect the gastrointestinal (GI) tract, liver, and pancreas, although several tumors with prominent "pink" (eosinophilic) cytoplasm are recognizable in the digestive tract. From a practical point of view, interest in the former lesions is limited to the differential diagnosis of a primary neoplasm versus metastatic deposits of malignant oncocytic tumors from other sites. This article briefly reviews the diagnostic features and clinical significance of the currently known tumors of the alimentary tract displaying a prominent oxyphilic character. PMID- 10452579 TI - Renal tumors exhibiting granular cytoplasm. AB - Renal epithelial neoplasms consist of a group of distinct genetic and clinical entities that occasionally have overlapping morphological features. Pronounced cytoplasmic granularity or eosinophilia may be seen in a number of tumor types, including conventional (clear-cell) carcinomas, papillary carcinomas, chromphobe carcinoma, collecting duct carcinomas, and oncocytomas. Mesenchymal neoplasms such as angiomyolipomas as well as metastatic lesions such as malignant melanoma may have marked epithelial features and cytoplasmic granularity, thus mimicking a renal epithelial tumor. The same can be said for adrenal cortical neoplasms, which sometimes may be confused clinically, radiographically, and pathologically with a renal neoplasm. Close attention to morphological and cytologic detail will solve the differential diagnosis in the majority of cases, although some will require ancillary studies such as histochemistry, immunohistochemistry, and election microscopy. In a small percentage of cases molecular genetic studies are required to properly classify the tumor. PMID- 10452580 TI - Oxyphilic tumors of the female and male genital tracts. AB - A great variety of oxyphilic tumors and tumor-like lesions occur in the female and male genital tracts, particularly in the former. The ovary is the site of a wider range of oxyphilic tumors than any other organ. Not only are its most highly specialized steroid hormone-secreting tumors and tumor-like lesions often completely or predominantly oxyphilic, but occasionally its epithelial cancers have a large component of oxyphilic cells. Several rare germ-cell tumors can also be completely or extensively oxyphilic and, being a common site of metastasis, the ovary can harbor a number of metastatic oxyphilic tumors. The lower female genital tract shares some of the problems in differential diagnosis with the ovary, and can be involved by a variety of mesenchymal oxyphilic tumors as well. In the male genital tract, the testis may be the site of a number of the same oxyphilic tumor types encountered in the ovary, but much less frequently, partly because the testis is an uncommon site of metastatic spread. PMID- 10452581 TI - Tumors of the skin composed of large cells with abundant eosinophilic cytoplasm. AB - Tumors of the skin composed of large epithelioid cells displaying abundant smooth or granular eosinophilic cytoplasm are relatively rare, yet may constitute a source of diagnostic difficulties for pathologists. Lesions derived from various skin elements are represented in this group, including tumors originating from adnexal structures and connective tissue elements of the dermis. The large, pink cell or epithelioid appearance of the tumor cells may be the result of attempts to recapitulate the phenotype of a cell line normally characterized by such morphological appearance, such as apocrine epithelium, or may result from secondary changes in the cytoplasm of the tumor cells, such as accumulation of autophagosomes in granular cell tumors. The characteristic pathological features of this group of tumors is the subject of this review, with special emphasis on their differential diagnosis and the morphological features that may be of help for practicing pathologists to arrive at a correct diagnosis. PMID- 10452582 TI - Tumors of the soft tissues composed of large eosinophilic cells. AB - Soft tissue neoplasms composed of large eosinophilic cells include benign and malignant tumors with different degrees of biological aggressiveness. The main histotypes discussed in this review are the heterogeneous group of benign and malignant granular cell tumors with neural and non-neural differentiation, alveolar soft part sarcomas, rhabdomyomas, and rhabdomyosarcomas. The salient anatomic, clinical, morphological, and immunophenotypic features in differential diagnosis with metastatic melanomas, carcinomas, and paragangliomas are discussed separately for each histotype. PMID- 10452583 TI - Oncocytic lesions of the neuroendocrine system. AB - Oncocytic tumors are rare tumors that have been described in many organs throughout the body, mainly in salivary, parathyroid, thyroid, and adrenal glands and in the kidney. A wide spectrum of benign and malignant oncocytic tumors can arise in various organs of the neuroendocrine system. They are usually defined as tumors that are composed predominantly or exclusively of oncocytic cells. A majority of these tumors are benign; however, in some endocrine organs such as the thyroid the diagnosis of oncocytic/Hurthle cell carcinoma portends a more aggressive clinical course than non-Hurthle cell follicular carcinoma. Therefore, it is important to follow strict criteria to diagnose and differentiate between benign and malignant oncocytic tumors. In this review we will discuss the clinicopathologic features of various oncocytic tumors of the endocrine glands. PMID- 10452584 TI - A pilot study demonstrating clinical benefit from intralesional interferon alpha 2B in the treatment of Peyronie's disease. AB - Intralesional therapy is a less invasive method for the treatment of Peyronie's disease. The objective of this study was to evaluate intralesional injections of interferon alpha 2B (IFN-alpha-2B) as an effective alternative to the surgical treatment of Peyronie's disease. Twenty-one patients with Peyronie's disease were evaluated by use of penile duplex Doppler ultrasonography for cavernosal blood flows, degree of penile curvature, and plaque size. A questionnaire was given to all patients to assess sexual function. Each patient then received biweekly intralesional injections of 1 x 10(6) units of IFN-alpha-2B in 10 ml of normal saline over a period of 6 months. There was no placebo control group in this study. At the conclusion of the study, penile duplex Doppler imaging was repeated. A questionnaire was completed by all patients to assess changes in sexual function after treatment. Twenty patients completed the study, with all men reporting subjective softening of their plaques. Nine of 10 patients initially reporting penile pain with erection (90%) had resolution of their phallalgia while on study protocol. Thirteen patients (65%) had significant improvement in curvature, ranging from 20 to 90%. Seventeen patients (85%) demonstrated an objective 10 to 80% decrease in plaque size. Biweekly intralesional injections of Peyronie's plaques with IFN-alpha-2B resulted in a significant improvement in penile curvature, diminished pain, and reduced plaque size, and resulted in a subjective improvement in sexual function. PMID- 10452585 TI - The use of percent free PSA for early detection of prostate cancer. PMID- 10452586 TI - Bioethics and law forum. PMID- 10452587 TI - Live sperm, dead bodies. PMID- 10452588 TI - The quagmire of postmortem sperm acquisition. PMID- 10452589 TI - Attributes of fertile spermatozoa: an update. PMID- 10452590 TI - Prostatic growth rate determined from MRI data: age-related longitudinal changes. AB - Men with prostatic enlargement are at highest risk of developing symptomatic lower urinary tract symptoms (LUTS) and related outcomes, such as acute urinary retention. The study of prostatic growth rate can identify the age range at which prostate growth peaks. Evaluation of the natural course of prostate growth requires repeated intraindividual volume measurements at time intervals sufficient to document growth. Our objective was to examine age-stratified prostate growth rates from men taking part in a longitudinal study of aging using magnetic resonance imaging (MRI) of the prostate. Sixty-four men (ages 30-71 years) enrolled in the Baltimore Longitudinal Study of Aging (BLSA) who had T2 pelvic MRIs taken approximately every 2 years were studied. Men were age stratified into four groups: <45, 45-55, 56-65, and >65 years old. Whole prostate and central gland (anatomically referred to as the transition zone) volumes were determined from the MRI images by a semi-automated image analysis program. Peripheral gland volumes were calculated as the difference between whole prostate and central gland volumes. Growth rates (cc per year) were calculated as change in volume divided by the time interval. On the basis of measurements from the T2 images (n = 128), we observed a linear trend between prostate volume and age. The overall prostate growth rate was 2.36 +/- 3.52 cc per year. Age-stratified growth rates revealed that prostate growth increased with age, peaked at 4.15 +/- 4.98 cc/year for the 56-65-year-old age group and then declined rapidly for the older aged men. The central gland growth rates followed a trend similar to total prostate volume. These data suggest that there is an age-related increase in prostate growth rate that peaks in men ages 56-65 and then declines. Identification of this trend in prostate growth may aid physicians in targeting men for early diagnosis of LUTS and for possible early intervention. Future studies with a larger sample size are necessary to substantiate these findings. PMID- 10452591 TI - Origin of an 84-kDa protein with ABH blood-typing antigen activity in human seminal plasma. AB - In order to investigate the origin of an 84-kDa protein with ABH blood-typing antigen activity (p 84) and its concentration in human seminal plasma, a monoclonal antibody (mAb p 84) was produced. The protein was recognized in breast milk as well as in seminal plasma by an indirect, enzyme-linked immunosorbent assay (ELISA) using this mAb. mAb p 84 identified 84-kDa and 83-kDa forms of the protein in seminal plasma and breast milk, respectively, on immunoblotting. The mean concentration of p 84 in seminal plasma was 949 microg/ml (n = 54 subjects). There was no significant difference in the concentration of p 84 between individuals who secreted (Se) or did not secrete (non-se) the ABH antigen into their seminal plasma, nor were there any significant correlations between the concentration of p 84 and the total seminal protein concentration. An immunohistochemical study using mAb p 84 with light microscopic detection showed that p 84 was located in the cytoplasm of the inner layer of pseudostratified cuboidal epithelial cells of the seminal vesicles, but no immunoreactivity was found in the prostate. These data indicate that p 84 originates from a single tissue, the seminal vesicles, and suggest that p 84 is an ABH epitope-bearing protein that has not previously been identified but possesses some immunological properties similar to those of lactotransferrin. PMID- 10452592 TI - Temporal relation between leptin and various indices of sexual maturation in the male rat. AB - Recent studies in humans and rhesus monkeys have suggested the possibility that the adipose tissue hormone leptin has a stimulatory and/or permissive effect on the onset of puberty in the male. We evaluated this hypothesis by measuring leptin in groups of male rats between the ages of 26 days and 96 days. A statistically significant positive correlation was present between serum leptin and age, body weight, prostate, seminal vesicle, and testes weight (both absolute and as a function of body weight). A statistically significant negative correlation was present between leptin and serum FSH and alpha-inhibin. There was not a statistically significant correlation between leptin and testosterone or LH. There was a statistically significant increase in the serum leptin concentrations at day 47. This rise was coincident with the peripubertal growth spurt in the secondary sexual organs and the peripubertal testosterone rise but occurred after the prepubertal rise in testicular weight, the appearance of elongating spermatids in the testes, and the start of the decline in FSH. In animals in which the peripubertal testosterone rise was delayed by the administration of EDS, serum leptin showed statistically significant differences from control. These data do not support the hypothesis that leptin provides a trigger for the onset of puberty in the male rat. They do suggest that leptin may be involved in the secondary sexual organ growth spurt and are consistent with the hypothesis that testosterone stimulates leptin synthesis during puberty. PMID- 10452593 TI - Comparison of fresh and cryopreserved human sperm attachment to bovine oviduct (uterine tube) epithelial cells in vitro. AB - Formation of a prefertilization sperm reservoir in mammals is thought to occur via sperm cell attachment to fallopian tube or oviduct epithelial cells (OEC). Recent data suggests that such an interaction also occurs for human sperm in the fallopian tube. We have previously validated an in vitro sperm-OEC coculture system utilizing bovine OEC monolayers to study postejaculatory human sperm physiology. This study was done to evaluate aspects of human sperm attachment to OEC in coculture and to determine if such attachment and subsequent sperm survival differ between fresh and cryopreserved human sperm. In experiment 1, aliquots of fresh (n = 4) or cryopreserved sperm (n = 3) from normospermic donors were placed into coculture with OEC monolayers at dilutions ranging from 2 x 10(5) to 15 x 10(6) sperm per well. Numbers of each type of sperm attaching to OEC at each concentration were determined. In experiment 2, fresh and cryopreserved sperm from the same donors (n = 4) were put into OEC coculture to observe numbers attaching and subsequent survival time for each sperm type. Sperm attachment to OEC occurred in a linear, dose-dependent manner for fresh and cryopreserved sperm in experiment 1, both as a function of total sperm numbers and as a function of numbers of motile sperm applied (R2 > or = 0.79). However, cryopreserved sperm attached to the OEC at a slower rate than fresh (as a function of the average increase in the number of sperm attaching per unit increase in the number of sperm applied; P < 0.05), with an overall lower percentage of the total and motile sperm applied attaching to OEC (P < 0.01) for cryopreserved versus fresh sperm. Fewer cryopreserved sperm also attached to the OEC, as compared with fresh sperm, in experiment 2 (P < 0.05), even after correcting for motility differences between the sperm types. Sperm survival time in coculture was also decreased for cryopreserved sperm as compared with fresh sperm (P = 0.005). Understanding the kinetics of sperm and OEC interactions may be useful for developing improved cryopreservation protocols or bioassays of sperm function. PMID- 10452594 TI - An improved mouse sperm-oocyte plasmalemma binding assay: studies on characteristics of sperm binding in medium with or without glucose. AB - Sperm binding to the oocyte plasmalemma is crucial to subsequent steps in fertilization. However, the usual in vitro assay for sperm-oocyte binding does not distinguish between nonspecific attachment and specific binding leading to fusion and penetration. Since zona pellucida-free pronuclear zygotes should not bind sperm (because of the block to polyspermy at the level of the oocyte membrane), a procedure has been developed to remove virtually all sperm from zona free pronuclear zygotes (2PNZ). After six washes with a 90-microm-bore pipette, there were 0.5 +/- 0.2 sperm/2PNZ (n = 83). Therefore, these washing conditions were used to define sperm-oocyte binding. The relationship of binding to oocyte penetration was determined for outbred mouse oocytes coincubated in complete medium with (B6 x 129)F1 hybrid sperm (10(7)/ml). Binding was maximal (29 +/- 5 sperm per oocyte) during the first 30 minutes but decreased significantly to 4.6 +/- 1.4 by 60 minutes of coincubation (over 10 trials). Oocyte penetration was 99 +/- 1% by 30 minutes, while the number of decondensed sperm nuclei per oocyte increased significantly to 7.5 +/- 0.6 at 60 minutes. These data suggest that the block to polyspermy involves detachment of bound sperm. Similar coincubations were carried out in medium without glucose (NoG), as this medium has been reported to inhibit fusion without affecting binding. However, binding was only 11 +/- 2 at 30 minutes but increased to 25 +/- 4 at 60 minutes, suggesting that binding was retarded in NoG. When gametes were coincubated in NoG for 5 hours, about half of the oocytes were penetrated, suggesting that the lack of glucose did not inhibit fusion but instead delayed it. These data suggest that if sperm binding is to be determined in complete medium, the time of the block to polyspermy should be determined prior to selecting the appropriate time to assay binding. Furthermore, using the same coincubation period for the binding assay in control and treated sperm may not be appropriate if the treatment alters the time of maximal binding. PMID- 10452595 TI - Reproductive aging in the Brown Norway rat is characterized by accelerated germ cell apoptosis and is not altered by luteinizing hormone replacement. AB - Reproductive aging in the male Brown Norway (BN) rat is characterized by decreased Leydig cell steroidogenesis associated with seminiferous tubule dysfunction. This could be a result of a combination of a primary testicular defect and a secondary hypothalamic pituitary dysfunction. In the present study, we determined in the BN rat whether germ cell loss occurred via apoptosis. We then defined the age of onset of Leydig cell dysfunction and germ cell loss and examined whether chronic luteinizing hormone (LH) replacement would delay or prevent reproductive aging. Plasma hormone levels, testicular sperm concentrations, and germ cell apoptosis were studied in 6, 9, 12, 15, 18, and 21 month-old BN rats. Beginning at 15 months, testicular weight, sperm concentration, total sperm counts, plasma testosterone, LH, and inhibin decreased, whereas the proportion of regressed testes and plasma follicle stimulating hormone (FSH) levels increased with aging. Accelerated germ cell apoptosis involving spermatogonia, preleptotene and pachytene spermatocytes, and spermatids was evident in some tubules of the relatively normal testes from 21 month-old rats. In the regressed testes, complete cessation of spermatogenesis occurred. The apoptotic index was higher in the testes of old (21-month-old) rats in particular at stages XII-XIV when compared with younger animals. Chronic LH replacement (0.5 microg i.p. twice per day) administered to 15-month-old BN rats for 6 months did not alter plasma hormone levels, testes weight, sperm concentration or content, or the germ cell apoptotic index. In the control group, 3 out of 10 testes were regressed, whereas in the LH-replaced group, only 1 out of 12 testes was regressed. We show in this study that early reproductive aging in the BN rat began at around 15 months. Germ cell loss associated with aging occurs via apoptosis. Replacement therapy with LH for 6 months does not decrease or delay the testicular dysfunction associated with aging. It is unlikely that hypothalamic-pituitary dysregulation is the major cause of testicular aging. PMID- 10452596 TI - Potential daily sperm production, Sertoli cell number, and seminiferous tubule characteristics in beef bulls fed conventional or gossypol-enriched diets. AB - The objective of this study was to determine the amount of germ cell degeneration throughout spermatogenesis in the bull and to determine if gossypol-enriched diets enhance that degeneration. Twenty-six adult beef bulls in two different sets of treatment groups were chosen for the study. The first set of bulls were broken into three groups, receiving soybean meal (<0.03 g gossypol per bull, per day), cottonseed meal (1.8 g gossypol per bull, per day), or whole cottonseed (16 g gossypol per bull per day). The second set of bulls were fed 4 g of gossypol per day for 180 days and then were placed in their respective treatment groups of 0 g or 4 g of gossypol per day for 72 days to evaluate the effect of continued gossypol treatment versus stopping treatment. Hence, one group received gossypol for 180 days, followed by withdrawal for 72 days. The treated group received gossypol up to castration at 250 days. In both sets of bulls, significant germ cell degeneration occurred between B spermatogonia and primary spermatocytes (i.e., at the end of mitosis and into the beginning of meiosis), with no subsequent degeneration throughout the rest of spermatogenesis. For both sets of bulls, the gossypol-enriched diet fed had no effect on the nuclear volume or number per gram parenchyma of any germ cell or Sertoli cell. Therefore, gossypol did not augment the amount of germ cell degeneration that occurred between B spermatogonia and homogenate-resistant (mature) spermatids in either set of bulls. Gossypol-enriched diets also had no effect on volume density, diameter, luminal diameter, or basement membrane thickness of seminiferous tubules for the first set of bulls. The second set of bulls, with continued diets enriched with gossypol, had a larger number of B spermatogonia per gram than did bulls removed from the gossypol-enriched diet. In conclusion, the bull did experience significant degeneration of germ cells between B spermatogonia and pachytene primary spermatocytes, with no subsequent loss throughout the rest of spermatogenesis, and the gossypol-enriched diet did not further enhance germ cell degeneration. PMID- 10452597 TI - Production of endothelin by canine prostatic epithelial cells and its stimulatory effects on their growth. AB - The current study has been designed to explore expressions of endothelin (ET) receptors and ETs in the canine prostate and the effect of ETs on canine prostatic epithelial cells. ET receptors were characterized by biotinylated ET-1 binding in frozen sections of the prostate. Canine prostatic epithelial cells in primary culture were used for demonstration of ET-1 expression by reverse-phase HPLC coupled with radioimmunoassay and Northern blotting and were subjected to growth assay. Biotinylated ET-1 binding was localized in the epithelial component, and the binding was also blocked with an antagonist specific for ET(B) subtype receptors. ET-1 and ET-3 stimulated canine prostatic epithelial cell growth in vitro. The effect was also reversed in the presence of an antagonist specific for ET(B) subtype receptors. Elution profile of epithelial cell culture medium revealed two peaks, corresponding to ET-1 and big ET-1. Epithelial cells in culture expressed pre-pro-ET-1 mRNA. Canine prostatic epithelial cells expressed ET(B) receptors and ET-1. It appears most likely that the expressed ET 1 acts as an autocrine/paracrine proliferative factor on canine prostatic epithelial cells via ET(B) receptors. PMID- 10452598 TI - Acrosome reaction in dog sperm is induced by a membrane-localized progesterone receptor. AB - The aim of this study was to investigate whether the dog sperm acrosome reaction can be induced by progesterone and whether the action of progesterone is mediated by binding of progesterone to a receptor on the sperm plasma. Progesterone-BSA conjugate labeled with fluorescein isothiocyanate (P-BSA-FITC) in combination with a vital stain, ethidium homodimer, was applied to visualize the presence of the progesterone receptor on living spermatozoa. Ten mM progesterone increased the acrosome reaction in viable spermatozoa over time from 3 +/- 1% at 0 hours to 69 +/- 8% at 6 hours (six dogs). In freshly ejaculated sperm from six dogs, P-BSA FITC staining was observed in 13 +/- 1% of the viable, acrosome-intact cells, as characterized by bright fluorescence over the entire apical region. The proportion of P-BSA-FITC-stained, viable, acrosome-intact cells increased to 84 +/- 11% following 7 hours incubation in a low-calcium medium. In contrast, the majority (72 +/- 3%) of fresh epididymal sperm already demonstrated bright P-BSA FITC staining. Apparently, epididymal spermatozoa already possess the progesterone receptor. The receptor is masked at ejaculation and subsequently gradually exposed. PMID- 10452599 TI - HLA-DR antigen and HLA-DRB1 genotyping with nonobstructive azoospermia in Japan. AB - We previously reported that the HLA-A33, -B13, and -B44 antigens, which are major histocompatibility complex class I molecules, are involved in the susceptibility of nonobstructive azoospermia in Japanese men. In this report, HLA-DR antigens, which are class II molecules, are investigated by advanced DNA typing in addition to classical serological typing to study a more complex genotype of HLA-DRB2. Genotyping was performed by the polymerase chain reaction-sequence-specific primer (PCR-SSP) method of analysis and/or by a commercial rapid assay based on the polymerase chain reaction (PCR), followed by reverse dot-blot hybridization of PCR products (the Inno-LiPA assay). The allele frequencies of the HLA-DR13 antigen and the -DRB1*1302 allele were significantly higher in Japanese subjects with nonobstructive azoospermia compared with a control group of healthy Japanese men, and these alleles were associated with relative risks for nonobstructive azoospermia of 4.2 and 4.9, respectively. If we suppose this strong linkage to both HLA class I and II antigens is due to linkage disequilibrium, it may suggest the existence of a novel gene involved in spermatogenesis in the class III region, which is located between the class I and class II regions and contains several genes other than HLA. PMID- 10452600 TI - Spermatic granulomata are experimentally induced in epididymides of mice receiving high-dose testosterone implants. I. A light-microscopical study. AB - It is known that a spermatic granuloma is induced by the inflammatory reaction following leakage of spermatozoa outside the germ cell ducts and is the main clinical complication of vasectomy. In the present study, we found that spermatic granulomata were experimentally induced in the epididymides of mice treated with high-dose testosterone. Testosterone powder (0.02, 0.2, or 2 mg per gram body weight) was implanted into ICR male mice, which were then killed from 7 to 63 days after the treatment for histological examination at the light-microscopic level. The results showed that the testis exhibited little or no degenerative change; however, the epididymides were frequently affected by spermatic granulomata after day 35 in mice implanted with high-dose testosterone (2 but not 0.2 or 0.02 mg per gram body weight). Observation of the early histological changes revealed that the ductal epithelium of the epididymides became vacuolated around day 25. Thereafter, the basement membrane of the epididymal ducts was ruptured after day 30, followed by leakage of spermatozoa into the adjacent interstitial tissue. The extravasated spermatozoa were then surrounded by macrophages (= epithelioid cells) and lymphocytes, resulting in the formation of a spermatic granuloma. In contrast, other mice treated with the same dose of deoxycorticosterone or estradiol did not show the induction of spermatic granulomata. Therefore, this study demonstrated that a spermatic granuloma is specifically formed in the epididymis by testosterone and that the lesion is started by vacuolation of the epididymal duct epithelium. PMID- 10452601 TI - Effects of liver disease and transplantation on the human prostate. AB - A major determinant of late-life prostate diseases is hormonal exposure during earlier life, but the effects of androgens in midlife on the human prostate have been little studied. In order to identify hormonal effects on the prostate during the long latent period of midlife, we studied the effects of chronic androgen deficiency on the prostate during midlife by examining men with severe liver disease before and after liver transplantation. Patients (n = 15, median 57, range 38-65 years old) with severe liver disease but no known prostate disease being evaluated for liver transplantation underwent 21 prostate ultrasound studies, 12 prior to and 9 after liver transplantation, with six men undergoing both studies. Controls were 42 prostate ultrasound studies (2:1 matching) from age-matched healthy men. Total- and central-prostate volumes were measured with a 7.5-MHz biplane transducer planimetrically at 2.5-mm intervals with a stepper device from base to apex of the prostate. Overall, total- and central-prostate volumes were not significantly different between patients with chronic liver disease before and after liver transplantation and age-matched healthy controls. This appeared to be due to a bimodal distribution, with most men (12 men, 17 studies) having smaller prostate volumes and a minority (3 men, 4 studies) having previously undiagnosed, macroscopic, benign prostatic hyperplasia. The reduction in prostate volume prior to transplantation was significantly correlated with severity of liver disease (Child-Pugh score). Before liver transplantation, prostate-specific antigen (PSA) concentrations were significantly lower and prostatic acid phosphatase increased, and both were normalized after liver transplantation. Plasma testosterone concentrations were decreased before transplantation and remained low after transplantation. Sex hormone-binding globulin level was significantly elevated before and reduced to subnormal after liver transplantation. Estradiol concentrations were unchanged by liver disease or transplantation. We conclude that prostate volumes, particularly that of the central zone, are usually reduced by the functional androgen deficiency of chronic liver disease and tend to be restored toward normal by liver transplantation, depending on the degree of rectification of circulating plasma testosterone concentrations. Prostate glands with established benign prostatic hyperplasia may be less responsive to these hormonal changes. PMID- 10452603 TI - Distinguishing between sequential and nonsequentially folded proteins: implications for folding and misfolding. AB - We describe here an algorithm for distinguishing sequential from nonsequentially folding proteins. Several experiments have recently suggested that most of the proteins that are synthesized in the eukaryotic cell may fold sequentially. This proposed folding mechanism in vivo is particularly advantageous to the organism. In the absence of chaperones, the probability that a sequentially folding protein will misfold is reduced significantly. The problem we address here is devising a procedure that would differentiate between the two types of folding patterns. Footprints of sequential folding may be found in structures where consecutive fragments of the chain interact with each other. In such cases, the folding complexity may be viewed as being lower. On the other hand, higher folding complexity suggests that at least a portion of the polypeptide backbone folds back upon itself to form three-dimensional (3D) interactions with noncontiguous portion(s) of the chain. Hence, we look at the mechanism of folding of the molecule via analysis of its complexity, that is, through the 3D interactions formed by contiguous segments on the polypeptide chain. To computationally splice the structure into consecutively interacting fragments, we either cut it into compact hydrophobic folding units or into a set of hypothetical, transient, highly populated, contiguous fragments ("building blocks" of the structure). In sequential folding, successive building blocks interact with each other from the amino to the carboxy terminus of the polypeptide chain. Consequently, the results of the parsing differentiate between sequentially vs. nonsequentially folded chains. The automated assessment of the folding complexity provides insight into both the likelihood of misfolding and the kinetic folding rate of the given protein. In terms of the funnel free energy landscape theory, a protein that truly follows the mechanism of sequential folding, in principle, encounters smoother free energy barriers. A simple sequentially folded protein should, therefore, be less error prone and fold faster than a protein with a complex folding pattern. PMID- 10452602 TI - The hydrogen exchange core and protein folding. AB - A database of hydrogen-deuterium exchange results has been compiled for proteins for which there are published rates of out-exchange in the native state, protection against exchange during folding, and out-exchange in partially folded forms. The question of whether the slow exchange core is the folding core (Woodward C, 1993, Trends Biochem Sci 18:359-360) is reexamined in a detailed comparison of the specific amide protons (NHs) and the elements of secondary structure on which they are located. For each pulsed exchange or competition experiment, probe NHs are shown explicitly; the large number and broad distribution of probe NHs support the validity of comparing out-exchange with pulsed-exchange/competition experiments. There is a strong tendency for the same elements of secondary structure to carry NHs most protected in the native state, NHs first protected during folding, and NHs most protected in partially folded species. There is not a one-to-one correspondence of individual NHs. Proteins for which there are published data for native state out-exchange and theta values are also reviewed. The elements of secondary structure containing the slowest exchanging NHs in native proteins tend to contain side chains with high theta values or be connected to a turn/loop with high theta values. A definition for a protein core is proposed, and the implications for protein folding are discussed. Apparently, during folding and in the native state, nonlocal interactions between core sequences are favored more than other possible nonlocal interactions. Other studies of partially folded bovine pancreatic trypsin inhibitor (Barbar E, Barany G, Woodward C, 1995, Biochemistry 34:11423-11434; Barber E, Hare M, Daragan V, Barany G, Woodward C, 1998, Biochemistry 37:7822-7833), suggest that developing cores have site-specific energy barriers between microstates, one disordered, and the other(s) more ordered. PMID- 10452604 TI - The disulfide-coupled folding pathway of apamin as derived from diselenide quenched analogs and intermediates. AB - The sequence of apamin, an 18 residue bee venom toxin, encloses all the information required for the correct disulfide-coupled folding into the cystine stabilized alpha-helical motif. Three apamin analogs, each containing a pair of selenocysteine residues replacing the related cysteines, were synthesized to mimic the three possible apamin isomers with two crossed, parallel, or consecutive disulfides, respectively. Refolding experiments clearly revealed that the redox potential of selenocysteine prevails over the sequence encoded structural information for proper folding of apamin. Thus, selenocysteine can be used as a new device to generate productive and nonproductive folding intermediates of peptides and proteins. In fact, disulfides are selectively reduced in presence of the diselenide and the conformational features derived from these intermediates as well as from the three-dimensional (3D) structures of the selenocysteine-containing analogs with their nonnatural networks of diselenide/disulfide bridges allowed to gain further insight into the subtle driving forces for the correct folding of apamin that mainly derive from local conformational preferences. PMID- 10452605 TI - Electrostatic forces involved in orienting Anabaena ferredoxin during binding to Anabaena ferredoxin:NADP+ reductase: site-specific mutagenesis, transient kinetic measurements, and electrostatic surface potentials. AB - Transient absorbance measurements following laser flash photolysis have been used to measure the rate constants for electron transfer (et) from reduced Anabaena ferredoxin (Fd) to wild-type and seven site-specific charge-reversal mutants of Anabaena ferredoxin:NADP+ reductase (FNR). These mutations have been designed to probe the importance of specific positively charged amino acid residues on the surface of the FNR molecule near the exposed edge of the FAD cofactor in the protein-protein interaction during et with Fd. The mutant proteins fall into two groups: overall, the K75E, R16E, and K72E mutants are most severely impaired in et, and the K138E, R264E, K290E, and K294E mutants are impaired to a lesser extent, although the degree of impairment varies with ionic strength. Binding constants for complex formation between the oxidized proteins and for the transient et complexes show that the severity of the alterations in et kinetics for the mutants correlate with decreased stabilities of the protein-protein complexes. Those mutated residues, which show the largest effects, are located in a region of the protein in which positive charge predominates, and charge reversals have large effects on the calculated local surface electrostatic potential. In contrast, K138, R264, K290, and K294 are located within or close to regions of intense negative potential, and therefore the introduction of additional negative charges have considerably smaller effects on the calculated surface potential. We attribute the relative changes in et kinetics and complex binding constants for these mutants to these characteristics of the surface charge distribution in FNR and conclude that the positively charged region of the FNR surface located in the vicinity of K75, R16, and K72 is especially important in the binding and orientation of Fd during electron transfer. PMID- 10452607 TI - The NTR module: domains of netrins, secreted frizzled related proteins, and type I procollagen C-proteinase enhancer protein are homologous with tissue inhibitors of metalloproteases. AB - Using homology search, structure prediction, and structural characterization methods we show that the C-terminal domains of (1) netrins, (2) complement proteins C3, C4, C5, (3) secreted frizzled-related proteins, and (4) type I procollagen C-proteinase enhancer proteins (PCOLCEs) are homologous with the N terminal domains of (5) tissue inhibitors of metalloproteinases (TIMPs). The proteins harboring this netrin module (NTR module) fulfill diverse biological roles ranging from axon guidance, regulation of Wnt signaling, to the control of the activity of metalloproteases. With the exception of TIMPs, it is not known at present what role the NTR modules play in these processes. In view of the fact that the NTR modules of TIMPs are involved in the inhibition of matrixin-type metalloproteases and that the NTR module of PCOLCEs is involved in the control of the activity of the astacin-type metalloprotease BMP1, it seems possible that interaction with metzincins could be a shared property of NTR modules and could be critical for the biological roles of the host proteins. PMID- 10452606 TI - The progressive development of structure and stability during the equilibrium folding of the alpha subunit of tryptophan synthase from Escherichia coli. AB - The urea-induced equilibrium unfolding of the alpha subunit of tryptophan synthase (alphaTS), a single domain alpha/beta barrel protein, displays a stable intermediate at approximately 3.2 M urea when monitored by absorbance and circular dichroism (CD) spectroscopy (Matthews CR, Crisanti MM, 1981, Biochemistry 20:784-792). The same experiment, monitored by one-dimensional proton NMR, shows another cooperative process between 5 and 9 M urea that involves His92 (Saab-Rincon G et al., 1993, Biochemistry 32:13,981-13,990). To further test and quantify the implied four-state model, N <--> I1 <--> I2 <--> U, the urea-induced equilibrium unfolding process was followed by tyrosine fluorescence total intensity, tyrosine fluorescence anisotropy and far-UV CD. All three techniques resolve the four stable states, and the transitions between them when the FL total intensity and CD spectroscopy data were analyzed by the singular value decomposition method. Relative to U, the stabilities of the N, I1, and I2 states are 15.4, 9.4, and 4.9 kcal mol(-1), respectively. I2 partially buries one or more of the seven tyrosines with a noticeable restriction of their motion; it also recovers approximately 6% of the native CD signal. This intermediate, which is known to be stabilized by the hydrophobic effect, appears to reflect the early coalescence of nonpolar side chains without significant organization of the backbone. I1 recovers an additional 43% of the CD signal, further sequesters tyrosine residues in nonpolar environments, and restricts their motion to an extent similar to N. The progressive development of a higher order structure as the denaturant concentration decreases implies a monotonic contraction in the ensemble of conformations that represent the U, I2, I1, and N states of alphaTS. PMID- 10452608 TI - Dissection of the protein G B1 domain binding site for human IgG Fc fragment. AB - The contribution to the free energy of binding of each of the residues forming the binding site for a human IgG Fc fragment on the surface of the B1 domain of protein G was determined by alanine-scanning mutagenesis. The interface between these two proteins is atypical in that it is smaller than usual, polar in character, and involves two well-defined "knobs-into-holes" interactions. The bulk of the free energy of binding is contributed by three central residues, which make hydrogen bonds across the interface. Of these, the most critical interaction is formed by Glu27, which acts as a charged knob on the surface of the B1 domain, inserting into a polar hole on the Fc fragment. A single alanine mutation of this residue virtually abolishes stable complex formation. Formation of a stable interface between these two proteins is therefore dominated by a small, polar "hot spot." PMID- 10452609 TI - Structure and dynamics of the fatty acid binding cavity in apo rat intestinal fatty acid binding protein. AB - The structure and dynamics of the fatty acid binding cavity in I-FABP (rat intestinal fatty acid binding protein) were analyzed. In the crystal structure of apo I-FABP, the probe occupied cavity volume and surface are 539+/-8 A3 and 428 A2, respectively (1.4 A probe). A total of 31 residues contact the cavity with their side chains. The side-chain cavity surface is partitioned according to the residue type as follows: 36-39% hydrophobic, 21-25% hydrophilic, and 37-43% neutral or ambivalent. Thus, the cavity surface is neither like a typical protein interior core, nor is like a typical protein external surface. All hydrophilic residues that contact the cavity-with the exception of Asp74-are clustered on the one side of the cavity. The cavity appears to expand its hydrophobic surface upon fatty acid binding on the side opposite to this hydrophilic patch. In holo I-FABP the fatty acid chain interactions with the hydrophilic side chains are mediated by water molecules. Molecular dynamics (MD) simulation of fully solvated apo I FABP showed global conformational changes of I-FABP, which resulted in a large, but seemingly transient, exposure of the cavity to the external solvent. The packing density of the side chains lining the cavity, studied by Voronoi volumes, showed the presence of two distinctive small hydrophobic cores. The MD simulation predicts significant structural perturbations of the cavity on the subnanosecond time scale, which are capable of facilitating exchange of I-FABP internal water. PMID- 10452610 TI - The alpha-subunit of protein prenyltransferases is a member of the tetratricopeptide repeat family. AB - Lipidation catalyzed by protein prenyltransferases is essential for the biological function of a number of eukaryotic proteins, many of which are involved in signal transduction and vesicular traffic regulation. Sequence similarity searches reveal that the alpha-subunit of protein prenyltransferases (PTalpha) is a member of the tetratricopeptide repeat (TPR) superfamily. This finding makes the three-dimensional structure of the rat protein farnesyltransferase the first structural model of a TPR protein interacting with its protein partner. Structural comparison of the two TPR domains in protein farnesyltransferase and protein phosphatase 5 indicates that variation in TPR consensus residues may affect protein binding specificity through altering the overall shape of the TPR superhelix. A general approach to evolutionary analysis of proteins with repetitive sequence motifs has been developed and applied to the protein prenyltransferases and other TPR proteins. The results suggest that all members in PTalpha family originated from a common multirepeat ancestor, while the common ancestor of PTalpha and other members of TPR superfamily is likely to be a single repeat protein. PMID- 10452611 TI - Escherichia coli maltose-binding protein is uncommonly effective at promoting the solubility of polypeptides to which it is fused. AB - Although it is usually possible to achieve a favorable yield of a recombinant protein in Escherichia coli, obtaining the protein in a soluble, biologically active form continues to be a major challenge. Sometimes this problem can be overcome by fusing an aggregation-prone polypeptide to a highly soluble partner. To study this phenomenon in greater detail, we compared the ability of three soluble fusion partners--maltose-binding protein (MBP), glutathione S-transferase (GST), and thioredoxin (TRX)--to inhibit the aggregation of six diverse proteins that normally accumulate in an insoluble form. Remarkably, we found that MBP is a far more effective solubilizing agent than the other two fusion partners. Moreover, we demonstrated that in some cases fusion to MBP can promote the proper folding of the attached protein into its biologically active conformation. Thus, MBP seems to be capable of functioning as a general molecular chaperone in the context of a fusion protein. A model is proposed to explain how MBP promotes the solubility and influences the folding of its fusion partners. PMID- 10452613 TI - Denaturant mediated unfolding of both native and molten globule states of maltose binding protein are accompanied by large deltaCp's. AB - Maltose binding protein (MBP) is a large, monomeric two domain protein containing 370 amino acids. In the absence of denaturant at neutral pH, the protein is in the native state, while at pH 3.0 it forms a molten globule. The molten globule lacks a tertiary circular dichroism signal but has secondary structure similar to that of the native state. The molten globule binds 8-anilino-1-naphthalene sulfonate (ANS). The unfolding thermodynamics of MBP at both pHs were measured by carrying out a series of isothermal urea melts at temperatures ranging from 274 329 K. At 298 K, values of deltaGdegrees , deltaCp, and Cm were 3.1+/-0.2 kcal mol(-1), 5.9+/-0.8 kcal mol(-1) K(-1) (15.9 cal (mol-residue)(-1) K(-1)), and 0.8 M, respectively, at pH 3.0 and 14.5+/-0.4 kcal mol(-1), 8.3+/-0.7 kcal mol(-1) K( 1) (22.4 kcal (mol-residue)(-1) K(-1)), and 3.3 M, respectively, at pH 7.1. Guanidine hydrochloride denaturation at pH 7.1 gave values of deltaGdegrees and deltaCp similar to those obtained with urea. The m values for denaturation are strongly temperature dependent, in contrast to what has been previously observed for small globular proteins. The value of deltaCp per mol-residue for the molten globule is comparable to corresponding values of deltaCp for the unfolding of typical globular proteins and suggests that it is a highly ordered structure, unlike molten globules of many small proteins. The value of deltaCp per mol residue for the unfolding of the native state is among the highest currently known for any protein. PMID- 10452612 TI - Folding propensities of synthetic peptide fragments covering the entire sequence of phage 434 Cro protein. AB - The phage 434 Cro protein, the N-terminal domain of its repressor (R1-69) and that of phage lambda (lambda6-85) constitute a group of small, monomeric, single domain folding units consisting of five helices with striking structural similarity. The intrinsic helix stabilities in lambda6-85 have been correlated to its rapid folding behavior, and a residual hydrophobic cluster found in R1-69 in 7 M urea has been proposed as a folding initiation site. To understand the early events in the folding of 434 Cro, and for comparison with R1-69 and lambda6-85, we examined the conformational behavior of five peptides covering the entire 434 Cro sequence in water, 40% (by volume) TFE/water, and 7 M urea solutions using CD and NMR. Each peptide corresponds to a helix and adjacent residues as identified in the native 434 Cro NMR and crystal structures. All are soluble and monomeric in the solution conditions examined except for the peptide corresponding to the 434 Cro helix 4, which has low water solubility. Helix formation is observed for the 434 Cro helix 1 and helix 2 peptides in water, for all the peptides in 40% TFE and for none in 7 M urea. NMR data indicate that the helix limits in the peptides are similar to those in the native protein helices. The number of side chain NOEs in water and TFE correlates with the helix content, and essentially none are observed in 7 M urea for any peptide, except that for helix 5, where a hydrophobic cluster may be present. The low intrinsic folding propensities of the five helices could account for the observed stability and folding behavior of 434 Cro and is, at least qualitatively, in accord with the results of the recently described diffusion-collision model incorporating intrinsic helix propensities. PMID- 10452614 TI - Effects of N-butyldeoxynojirimycin and the Lec3.2.8.1 mutant phenotype on N glycan processing in Chinese hamster ovary cells: application to glycoprotein crystallization. AB - Heterologous gene expression in either (1) the glycosylation-defective, mutant Chinese hamster ovary cell line, Lec3.2.8.1, or (2) the presence of the alpha glucosidase inhibitor, N-butyldeoxynojirimycin facilitates the trimming of N linked glycans of glycoproteins to single N-acetylglucosamine (GlcNAc) residues with endoglycosidase H (endo H). Both approaches are somewhat inefficient, however, with as little as 12% of the total protein being rendered fully endo H sensitive under these conditions. It is shown here that the combined effects of these approaches on the restriction of oligosaccharide processing are essentially additive, thereby allowing the production of glycoproteins that are essentially completely endo H-sensitive. The preparation of a soluble chimeric form of CD58, the ligand of the human T-cell surface recognition molecule CD2, illustrates the usefulness of the combined approach when expression levels are low or the deglycosylated protein is unstable at low pH. The endo H-treated chimera produced crystals of space group P3(1)21 or P3(2)21, and unit cell dimensions a = b = 116.4 A, c = 51.4 A alpha = beta = 90 degrees , gamma = 120 degrees , that diffract to a maximum resolution of 1.8 A. PMID- 10452615 TI - Drug resistance mutations can effect dimer stability of HIV-1 protease at neutral pH. AB - The monomer-dimer equilibrium for the human immunodeficiency virus type 1 (HIV-1) protease has been investigated under physiological conditions. Dimer dissociation at pH 7.0 was correlated with a loss in beta-sheet structure and a lower degree of ANS binding. An autolysis-resistant mutant, Q7K/L33I/L63I, was used to facilitate sedimentation equilibrium studies at neutral pH where the wild-type enzyme is typically unstable in the absence of bound inhibitor. The dimer dissociation constant (KD) of the triple mutant was 5.8 microM at pH 7.0 and was below the limit of measurement (approximately 100 nM) at pH 4.5. Similar studies using the catalytically inactive D25N mutant yielded a KD value of 1.0 microM at pH 7.0. These values differ significantly from a previously reported value of 23 nM obtained indirectly from inhibitor binding measurements (Darke et al., 1994). We show that the discrepancy may result from the thermodynamic linkage between the monomer-dimer and inhibitor binding equilibria. Under conditions where a significant degree of monomer is present, both substrates and competitive inhibitors will shift the equilibrium toward the dimer, resulting in apparent increases in dimer stability and decreases in ligand binding affinity. Sedimentation equilibrium studies were also carried out on several drug-resistant HIV-1 protease mutants: V82F, V82F/I84V, V82T/I84V, and L90M. All four mutants exhibited reduced dimer stability relative to the autolysis-resistant mutant at pH 7.0. Our results indicate that reductions in drug affinity may be due to the combined effects of mutations on both dimer stability and inhibitor binding. PMID- 10452617 TI - NMR study of Ni2+ binding to the H-N-H endonuclease domain of colicin E9. AB - Ni2+ affinity columns are widely used for protein purification, but they carry the risk that Ni2+ ions may bind to the protein, either adventitiously or at a physiologically important site. Dialysis against ethylenediaminetetraacetic acid (EDTA) is normally used to remove metal ions bound adventitiously to proteins; however, this approach does not always work. Here we report that a bacterial endonuclease, the DNase domain of colicin E9, binds Ni2+ acquired from Ni2+ affinity columns, and appears to bind [Ni(EDTA)(H2O)n]2- at low ionic strength. NMR was used to detect the presence of both Ni2+ coordinated to amino acid side chains and [Ni(EDTA)(H2O)N]2-. Dialysis against > or =0.2 M NaCl was required to remove the [Ni(EDTA)(H2O)n]2-. The NMR procedure we have used to characterize the presence of Ni2+ and [Ni(EDTA)(H2O)n]2- should be applicable to other proteins where there is the possibility of binding paramagnetic metal ions that are present to expedite protein purification. In the present case, the binding of Ni2+ seems likely to be physiologically relevant, and the NMR data complement recent X-ray crystallographic evidence concerning the number of histidine ligands to bound Ni2+. PMID- 10452616 TI - p53 Family members p63 and p73 are SAM domain-containing proteins. AB - Homologs of the tumor suppressor p53, called p63 and p73, have been identified. The p63 and p73 family members possess a domain structure similar to p53, but contain variable C-terminal extensions. We find that some of the C-terminal extensions contain Sterile Alpha Motif (SAM) domains. SAM domains are protein modules that are involved in protein-protein interactions. Consistent with this role, the C-terminal SAM domains of the p63 and p73 may regulate function by recruiting other protein effectors. PMID- 10452619 TI - Tropical infectious diseases in human immunodeficiency virus-infected patients. PMID- 10452618 TI - A superfamily of archaeal, bacterial, and eukaryotic proteins homologous to animal transglutaminases. AB - Computer analysis using profiles generated by the PSI-BLAST program identified a superfamily of proteins homologous to eukaryotic transglutaminases. The members of the new protein superfamily are found in all archaea, show a sporadic distribution among bacteria, and were detected also in eukaryotes, such as two yeast species and the nematode Caenorhabditis elegans. Sequence conservation in this superfamily primarily involves three motifs that center around conserved cysteine, histidine, and aspartate residues that form the catalytic triad in the structurally characterized transglutaminase, the human blood clotting factor XIIIa'. On the basis of the experimentally demonstrated activity of the Methanobacterium phage pseudomurein endoisopeptidase, it is proposed that many, if not all, microbial homologs of the transglutaminases are proteases and that the eukaryotic transglutaminases have evolved from an ancestral protease. PMID- 10452620 TI - Threats to global health and survival: the growing crises of tropical infectious diseases--our "unfinished agenda". AB - Health, one of our most unassailable human values, transcends all geographic, political, and cultural boundaries. The health problems of the rapidly growing 80% of the world's population that live in the tropical developing countries of Asia, Africa, and Latin America pose major threats to industrialized as well as developing regions. These threats can be divided into three areas, or three "E"s: (1) emerging, reemerging, and antimicrobial-resistant infections; (2) exploding populations without improved health; and (3) erosion of our humanity or leadership if we ignore the growing health problems of the poor. Our assessment of current trends in global population distribution and resource consumption; DALY calculations, causes, and distribution of global mortality and morbidity; and the misperceptions about and maldistribution of resources for health point to the critical importance of addressing tropical infectious diseases and global health for preservation of democracy and civilization as we know it. PMID- 10452621 TI - Photo quiz. Schistosoma haematobium. PMID- 10452623 TI - The role of Chlamydia pneumoniae in atherosclerosis. PMID- 10452622 TI - Low prevalence of Chlamydia pneumoniae in atherectomy specimens from patients with coronary heart disease. AB - Coronary atherectomy specimens from 50 patients with coronary heart disease were examined for the presence of Chlamydia pneumoniae by two different methods of polymerase chain reaction (PCR) and by in situ hybridization. C. pneumoniae DNA was detected by PCR in atherosclerotic plaques of four patients (8%). Two patients' coronary atheromas were positive, both by a single-step 16S rRNA-based PCR and by an omp1-based nested PCR. The other two patients' specimens were positive only by the nested PCR. In contrast, C. pneumoniae was not detected by in situ hybridization in any of the cardiovascular tissues tested. Of three patients with evidence of C. pneumoniae in coronary atheromas, two had an antibody titer of 1:32 and the third had no specific antibodies detectable. Results of this study demonstrate a low prevalence of C. pneumoniae DNA in coronary atheromas. These findings do not support the hypothesis that the organism plays a major role in atherogenesis. PMID- 10452624 TI - Safety and immunogenicity of Haemophilus influenzae-tetanus toxoid conjugate vaccine given separately or in combination with a three-component acellular pertussis vaccine combined with diphtheria and tetanus toxoids and inactivated poliovirus vaccine for the first four doses. AB - The purpose of this randomized, controlled trial was to assess the safety and immunogenicity of a three-component acellular pertussis vaccine combined with diphtheria and tetanus toxoids and inactivated poliovirus vaccine given either separately or combined as a single injection with a Haemophilus influenzae type b tetanus toxoid conjugate vaccine. A total of 180 infants were immunized at 2, 4, and 6 months of age; 129 were given a booster dose at 16-19 months of age. Vaccine-associated adverse events were similar whether the vaccines were combined as a single injection or given separately. There were no differences in levels of antibodies to Bordetella pertussis antigens (pertussis toxoid, filamentous hemagglutinin, and pertactin), diphtheria toxoid, or the three poliovirus types. The tetanus antitoxin level after the primary three-dose series was higher in recipients of the combined vaccine (2.37 IU/mL) than in recipients of the separate injections (1.32 IU/mL; two-sided P = .0001). In contrast, combined vaccine recipients had lower levels of antibody to H. influenzae type b polysaccharide after the third dose (1.57 microg/mL) than did those given separate injections (3.22 microg/mL; two-sided P = .0026). The antibody levels were not significantly different before or 1 month after the booster dose (32.9 microg/mL vs. 47.8 microg/mL, respectively; two-sided P = .07). We conclude that the vaccines were immunogenic and well tolerated. Despite lower levels of antibody to the H. influenzae type b polysaccharide after the primary three-dose series, mixing of the vaccines in a single syringe likely induced immunologic priming, as suggested by the high antibody levels after the booster dose. PMID- 10452625 TI - Screening for Chlamydia trachomatis infection in college women with a polymerase chain reaction assay. AB - This study sought to determine factors associated with chlamydial infection in a low-prevalence college health setting and to determine the testing characteristics of a polymerase chain reaction (PCR) assay for chlamydial infection (AMPLICOR chlamydia test; Roche Diagnostic Systems, Indianapolis) in this population. Young women (n = 1,149) at a university student health clinic underwent testing for cervical chlamydial infection by PCR assay and culture; the characteristics of women with and without chlamydial infection were compared. Chlamydial infection was diagnosed for 26 students (2.3%). The sensitivity and specificity of PCR assay and culture were 85% and 100% and 54% and 100%, respectively. Students with chlamydial infection were more likely to be 20 years of age or younger, have symptoms, report prior chlamydial infection or gonorrhea, report exposure to a sexually transmitted disease (STD), be black, or have cervical signs during examination; however, none of these were significant predictors for asymptomatic women. PCR assay detected significantly more cervical infections than did culture in this college student population. These data are consistent with recommendations for testing college women with symptoms, STD exposure, or age of younger than 25 years. PMID- 10452626 TI - Intravenous colistin as therapy for nosocomial infections caused by multidrug resistant Pseudomonas aeruginosa and Acinetobacter baumannii. AB - Sixty nosocomial infections caused by Pseudomonas aeruginosa and Acinetobacter baumannii resistant to aminoglycosides, cephalosporins, quinolones, penicillins, monobactams, and imipenem were treated with colistin (one patient had two infections that are included as two different cases). The infections were pneumonia (33% of patients), urinary tract infection (20%), primary bloodstream infection (15%), central nervous system infection (8%), peritonitis (7%), catheter-related infection (7%), and otitis media (2%). A good outcome occurred for 35 patients (58%), and three patients died within the first 48 hours of treatment. The poorest results were observed in cases of pneumonia: only five (25%) of 20 had a good outcome. A good outcome occurred for four of five patients with central nervous system infections, although no intrathecal treatment was given. The main adverse effect of treatment was renal failure; 27% of patients with initially normal renal function had renal failure, and renal function worsened in 58% of patients with abnormal baseline creatinine levels. Colistin may be a good therapeutic option for the treatment of severe infections caused by multidrug-resistant P. aeruginosa and A. baumannii. PMID- 10452627 TI - Overexpression of Fas/CD95 and Fas-induced apoptosis in a patient with idiopathic CD4+ T lymphocytopenia. AB - The mechanisms of apoptosis have become better understood, in part with the discovery of Fas/CD95. We report the case of a patient characterized by a decreased CD4+ T cell count and an overexpression of Fas/CD95 resulting in apoptosis. A 54-year-old man presented with disseminated Mycobacterium xenopi infection. Analysis showed CD4+ T lymphopenia. Tests for human immunodeficiency virus (HIV) types 1 and 2 were negative. We compared the patient with eight healthy controls and five HIV-infected patients in terms of the expression of Fas/CD95 and Fas-mediated apoptosis of peripheral T lymphocytes. The percent of CD95+ cells in lymphocytes was 98% for the patient, and the mean percent of CD95+ cells in lymphocytes +/- SD for HIV-infected patients and healthy controls was 75% +/- 16% and 36% +/- 26%, respectively. The patient had a high level of spontaneous apoptosis, and apoptotic cells were all identified as being CD4+ T cells. Monoclonal antibodies to CD95 dramatically increased apoptosis of CD4+ T cells exclusively. CD4+ T lymphopenia observed in our patient correlated with an overexpression of Fas together with spontaneous and Fas-induced apoptosis. PMID- 10452628 TI - Impact of use of multiple antimicrobials on changes in susceptibility of gram negative aerobes. AB - Evaluation of antimicrobial usage vs. susceptibility relationships typically involves single agents. However, susceptibility profiles may be affected by multiple drugs. From 1992 through 1996, we studied relationships between drug usage and the susceptibility (only susceptibility rates of > or = 70%) of Acinetobacter anitratus (baumannii), Enterobacter aerogenes, Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, and Serratia marcescens to 22 agents. Linear regression was used to assess usage of each agent vs. susceptibility to it and to all agents. Only relationships with a coefficient of determination of > or = 0.5 and a negative slope were evaluated and classified as increasing drug use and decreasing susceptibility (increasing D, decreasing %S) or decreasing drug use and increasing susceptibility (decreasing D, increasing %S). The mean numbers (range) of drugs associated with a change in susceptibility were 1.7 (0-14) and 0.6 (0 7), respectively, for increasing D, decreasing %S and decreasing D, increasing %S relationships. Multiple antimicrobials are associated with susceptibility to other drugs; thus, surveillance of these relationships should not be limited to single drugs. PMID- 10452629 TI - Evolution of vaginal Candida species recovered from human immunodeficiency virus infected women receiving fluconazole prophylaxis: the emergence of Candida glabrata? Terry Beirn Community Programs for Clinical Research in AIDS (CPCRA). AB - The effect of fluconazole prophylaxis on the vaginal flora of 323 human immunodeficiency virus-infected women was evaluated in a multicenter, randomized, double-blind, placebo-controlled trial. Women with CD4 cell counts of < or = 300/mm3 received either 200 mg of fluconazole per week or placebo. Vaginal surveillance cultures were performed every 3 months. After a follow-up of 29 months, Candida albicans was recovered from 53% of patients receiving fluconazole and 68% of patients assigned placebo. Fluconazole was associated with a 50% reduction in the odds of being colonized with C. albicans but with higher rates for non-albicans Candida species. Candida glabrata was recovered from 40 women assigned fluconazole and 29 assigned placebo (relative odds, 1.96; 95% confidence interval, 0.98-3.94). Fluconazole had an early and persistent effect on the vaginal mycoflora, with the emergence of C. glabrata vaginal colonization within the first 6 months. The effect of fluconazole prophylaxis can be attributed to the reduction in vaginal C. albicans colonization; however, C. glabrata colonization rapidly supervened. PMID- 10452630 TI - Chronic active hepatitis B exacerbations in human immunodeficiency virus-infected patients following development of resistance to or withdrawal of lamivudine. AB - Lamivudine is a nucleoside analog with activity against human immunodeficiency virus (HIV) and hepatitis B virus (HBV). Patients coinfected with HIV and HBV may have hepatitis flares when lamivudine therapy is discontinued or when resistance of HBV to lamivudine emerges. This retrospective, descriptive study conducted in three tertiary care medical centers describes patients coinfected with HIV type 1 and HBV who presented with a spectrum of clinical and subclinical hepatitic responses to lamivudine withdrawal or resistance. One patient had fulminant hepatic failure and a second patient had subclinical hepatitis when lamivudine therapy was discontinued and a more efficacious antiretroviral regimen was substituted. Three patients had flares of hepatitis after 13 to 18 months of lamivudine therapy. Lamivudine withdrawal or emergence of lamivudine-resistant mutants in patients coinfected with HIV and HBV may result in severe hepatitis. Clinicians caring for patients with coinfection with HIV and HBV should be aware of the possibility that a hepatitis B flare may occur in previously asymptomatic carrier patients. PMID- 10452631 TI - Interferon gamma (IFN-gamma) deficiency in generalized Epstein-Barr virus infection with interstitial lymphoid and granulomatous pneumonia, focal cerebral lesions, and genital ulcers: remission following IFN-gamma substitution therapy. AB - A 26-year-old previously healthy woman developed granulomatous pneumonitis, encephalitis, and genital ulceration during primary Epstein-Barr virus (EBV) infection. EBV DNA was demonstrated by polymerase chain reaction analysis of serum, lung tissue, and genital ulcer specimens. Serology verified primary EBV infection. The patient lacked lymphocytes cytotoxic to autologous EBV-transformed B lymphocytes. No spontaneous or in vitro EBV-induced interferon gamma (IFN gamma) production was evident in peripheral blood. The cells had normal IFN-gamma production when stimulated with Staphylococcus aureus exotoxin A. In the bone marrow and peripheral blood, the number of large granular CD56+ lymphocytes (natural killer cells) increased 39%-55%, but no CD4 or CD8 cell lymphocytosis was initially found. A partial clinical response was achieved with treatment with acyclovir, corticosteroids, and intravenous gamma-globulin. Because of persistent granulomatous central nervous system and lung involvement, subcutaneous IFN-gamma therapy was started but was discontinued after 3 months because of development of fever, pancytopenia, and hepatitis. This therapy initiated a complete clinical recovery, which occurred parallel to development of EBV-specific cytotoxic CD8+ T lymphocytes and normalization of natural killer cell lymphocytosis. These findings provide evidence for an EBV-induced lymphoproliferative disorder due to a T lymphocyte dysfunction associated with a selective lack of IFN-gamma synthesis. PMID- 10452632 TI - Conventional respiratory viruses recovered from immunocompromised patients: clinical considerations. AB - Lower respiratory tract infection is the most common complication in the immunocompromised patient. From January 1991 to December 1995, 785 consecutive patients with suspected respiratory tract infections were studied. One hundred ninety-nine viruses were isolated from 182 (23%) of 785 bronchoalveolar lavage fluid specimens. Cytomegalovirus was isolated from 131 patients, herpes simplex virus was recovered from 31, and conventional respiratory viruses (CRVs) were recovered from 36. There were 9 influenza A viruses, 2 influenza B viruses, 7 parainfluenza viruses, 5 respiratory syncytial viruses, 5 adenoviruses, 6 enteroviruses, and 3 rhinoviruses. We identified 22 patients from whom a CRV was the only microorganism recovered; 13 patients developed pneumonia, 10 had acute respiratory failure, 5 required support with mechanical ventilation, and 5 (23%) died. In conclusion, CRVs are frequent causes of respiratory illnesses and are associated with high rates of morbidity and mortality among immunocompromised patients. PMID- 10452633 TI - Itraconazole prophylaxis for fungal infections in patients with advanced human immunodeficiency virus infection: randomized, placebo-controlled, double-blind study. National Institute of Allergy and Infectious Diseases Mycoses Study Group. AB - In a prospective, randomized, double-blind trial, 149 patients with advanced human immunodeficiency virus (HIV) infection were randomized to receive itraconazole capsules (200 mg daily) and 146 to receive a matched placebo. Both groups were monitored for evidence of fungal infections. Baseline characteristics of the two groups were similar. Failure of prophylaxis occurred in 29 (19%) of the itraconazole recipients and 42 (29%) of the placebo recipients (P = .004; log rank test). There were 6 invasive fungal infections in the itraconazole group (4, histoplasmosis; 1, cryptococcosis; 1, aspergillosis) and 19 in the placebo group (10, histoplasmosis; 8, cryptococcosis; 1, aspergillosis) (P = .0007; log-rank test). Itraconazole significantly delayed time to onset of histoplasmosis (P = .03; log-rank test) and cryptococcosis (P = .0005; log-rank test). Prophylaxis failure due to recurrent or refractory mucosal candidiasis occurred with similar frequency in the two groups (itraconazole, 15%; placebo, 16%). A survival benefit was not demonstrated. Itraconazole generally was well tolerated. Primary prophylaxis with itraconazole capsules prevents histoplasmosis and cryptococcosis in patients with HIV infection. PMID- 10452634 TI - Use of corticosteroids in glomerulonephritis related to infective endocarditis: three cases and review. AB - We report the cases of three patients treated for infective endocarditis (IE) for whom corticosteroids were added to the antibiotic treatment. They all had clinical and biological evidence of immune-mediated glomerulonephritis. The microorganisms responsible for IE were Coxiella burnetii, Streptococcus bovis, and Cardiobacterium hominis. Median duration of IE before antimicrobial therapy was 7 months. In all patients, renal function deteriorated despite appropriate antimicrobial treatment for a mean duration of 16 days, but it improved after addition of corticosteroid therapy. All patients were cured of IE. A literature review revealed four additional cases of IE-related glomerulonephritis in which adjunctive immunosuppressive therapy was considered to be effective. Although corticosteroid therapy is generally not recommended for IE, it should be considered for patients whose renal dysfunction secondary to glomerulonephritis does not improve with appropriate antimicrobial treatment, especially if the duration of the illness is long. PMID- 10452635 TI - Reduction in the incidence of methicillin-resistant Staphylococcus aureus and ceftazidime-resistant Klebsiella pneumoniae following changes in a hospital antibiotic formulary. AB - In 1995, changes in our hospital formulary were made to limit an outbreak of vancomycin-resistant enterococci and resulted in decreased usage of cephalosporins, imipenem, clindamycin, and vancomycin and increased usage of beta lactam/beta-lactamase-inhibitor antibiotics. In this report, the effect of this formulary change on other resistant pathogens is described. Following the formulary change, there was a reduction in the monthly number (mean +/- SD) of patients with methicillin-resistant Staphylococcus aureus (from 21.9 +/- 8.1 to 17.2 +/- 7.2 patients/1,000 discharges; P = .03) and ceftazidime-resistant Klebsiella pneumoniae (from 8.6 +/- 4.3 to 5.7 +/- 4.0 patients/1,000 discharges; P = .02). However, there was an increase in the number of patients with cultures positive for cefotaxime-resistant Acinetobacter species (from 2.4 +/- 2.2 to 5.4 +/- 4.0 patients/1,000 discharges; P = .02). Altering an antibiotic formulary may be a possible mechanism to contain the spread of selected resistant pathogens. However, close surveillance is needed to detect the emergence of other resistant pathogens. PMID- 10452636 TI - A silver bullet for colonization and infection with methicillin-resistant Staphylococcus aureus still eludes us. PMID- 10452637 TI - Candidemia in cancer patients: a prospective, multicenter surveillance study by the Invasive Fungal Infection Group (IFIG) of the European Organization for Research and Treatment of Cancer (EORTC). AB - In a surveillance study of candidemia in cancer patients that was conducted by the European Organization for Research and Treatment of Cancer, 249 episodes were noted; Candida albicans was isolated in 70% (63) of the 90 cases involving patients with solid tumors (tumor patients) and in 36% (58) of the 159 involving those with hematologic disease (hematology patients). Neutropenia in tumor patients and acute leukemia and antifungal prophylaxis in hematology patients were significantly associated with non-albicans candidemia in a multivariate analysis. Overall 30-day mortality was 39% (97 of 249). In a univariate analysis, Candida glabrata was associated with the highest mortality rate (odds ratio, 2.66). Two multivariate analyses showed that mortality was associated with older age and severity of the underlying disease. Among hematology patients, additional factors associated with mortality were allogeneic bone marrow transplantation, septic shock, and lack of antifungal prophylaxis. PMID- 10452638 TI - A randomized, placebo-controlled study of rifabutin added to a regimen of clarithromycin and ethambutol for treatment of disseminated infection with Mycobacterium avium complex. AB - Current guidelines suggest that disseminated Mycobacterium avium complex (MAC) infection be treated with a macrolide plus ethambutol or rifabutin or both. From 1993 to 1996, 198 AIDS patients with MAC bacteremia participated in a prospective, placebo-controlled trial of clarithromycin (500 mg b.i.d.) plus ethambutol (1,200 mg/d), with or without rifabutin (300 mg/d). At 16 weeks, 63% of patients in the rifabutin group and 61% in the placebo group (P = .81) had responded bacteriologically. Changes in clinical symptoms and time to survival were similar in both groups. Development of clarithromycin resistance during therapy was similar in the two groups; of patients who had a bacteriologic response, however, only 1 of 44 (2%) receiving rifabutin developed clarithromycin resistance, vs. 6 of 42 (14%) in the placebo group (P = .055). Thus, rifabutin had no impact on bacteriologic response or survival but may protect against development of clarithromycin resistance in those who respond to therapy. PMID- 10452639 TI - Genital ulcers: etiology, clinical diagnosis, and associated human immunodeficiency virus infection in Kingston, Jamaica. AB - Individuals presenting consecutively with genital ulcers in Kingston, Jamaica, underwent serological testing for human immunodeficiency virus (HIV) infection, chlamydial infection, and syphilis. Ulcer material was analyzed by multiplex polymerase chain reaction (M-PCR) analysis. DNA from herpes simplex virus (HSV), Haemophilus ducreyi, and Treponema pallidum was detected in 158 (52.0%), 72 (23.7%), and 31 (10.2%) of 304 ulcer specimens. Of the 304 subjects, 67 (22%) were HIV-seropositive and 64 (21%) were T. pallidum-seroreactive. Granuloma inguinale was clinically diagnosed in nine (13.4%) of 67 ulcers negative by M-PCR analysis and in 12 (5.1%) of 237 ulcers positive by M-PCR analysis (P = .03). Lymphogranuloma venereum was clinically diagnosed in eight patients. Compared with M-PCR analysis, the sensitivity and specificity of a clinical diagnosis of syphilis, herpes, and chancroid were 67.7%, 53.8%, and 75% and 91.2%, 83.6%, and 75.4%, respectively. Reactive syphilis serology was 74% sensitive and 85% specific compared with M-PCR analysis. Reported contact with a prostitute in the preceding 3 months was associated with chancroid (P = .009), reactive syphilis serology (P = .011), and HIV infection (P = .007). The relatively poor accuracy of clinical and locally available laboratory diagnoses pleads for syndromic management of genital ulcers in Jamaica. Prevention efforts should be intensified. PMID- 10452640 TI - Clinical case definitions for Argentine hemorrhagic fever. AB - Argentine hemorrhagic fever (AHF) is a potentially lethal infection in Argentina. The case-fatality ratio is >15%, but treatment reduces the mortality rate to <1%. Diagnosis is based on clinical and laboratory criteria, but no case definition has been validated. A chart review was conducted for patients hospitalized with suspected AHF. Individuals with a fourfold rise in antibody titer were classified as cases. The combination of a platelet count of <100,000/mm3 and a white blood cell (WBC) count of <2,500/mm3 had a sensitivity and specificity of 87% and 88%, respectively, thus suggesting that the use of these criteria in a case definition would be helpful for epidemiological studies of AHF. The combination of a platelet count of <100,000/mm3 and a WBC count of <4,000/mm3 had a sensitivity of 100% and a specificity of 71%; the use of these criteria in a case definition should be helpful for screening patients for therapy with immune plasma in the region where AHF is endemic. PMID- 10452641 TI - Bordetella bronchiseptica infection in human immunodeficiency virus-infected patients. AB - Bordetella bronchiseptica is a pleomorphic gram-negative coccobacillus that commonly causes respiratory tract infections in dogs. We identified nine human immunodeficiency virus (HIV)-infected persons with culture-confirmed B. bronchiseptica infections (eight respiratory tract and one disseminated infection). The respiratory illnesses ranged in severity from mild upper respiratory tract infection to pneumonia. All nine patients had had at least one AIDS-defining condition before the B. bronchiseptica infection. Two patients had household contact with dogs before their illnesses, and one had household contact with cats. Infection due to B. bronchiseptica is uncommon in HIV-infected persons. Additional data are needed to fully define the spectrum of disease due to B. bronchiseptica infections and to evaluate the possibility that this infection may be acquired from pets. Treatment of B. bronchiseptica infection should be tailored to the patient and should be based on the results of susceptibility testing. PMID- 10452642 TI - Antibody response after a four-site intradermal booster vaccination with cell culture rabies vaccine. AB - The current World Health Organization recommendation for booster vaccination of previously immunized individuals with potential exposure to rabies is two doses of vaccine intramuscularly or intradermally on days 0 and 3. We report responses to two types of postexposure treatment of healthy individuals who had received preexposure rabies vaccination 1 year previously. Group A individuals received four intradermal doses (one-fifth of the diluent volume of vaccine per dose) on day 0, and group B individuals received two intramuscular doses on days 0 and 3. Immunogenicity of the two booster regimens was assessed by titrating the amount of neutralizing antibody (Nab). We found that the booster doses of vaccine produced remarkable responses in all subjects. Nab titers of > or = 0.5 IU/mL (acceptable antibody level for protection against rabies) were detected in all subjects on day 14, and they were shown to be consistently high 1 year after the booster vaccination. We also found that the Nab titers for group A were significantly higher (two- to eightfold) than those for group B on days 5, 14, 150, and 360 after the initial booster vaccination (P < .05). Our study shows that the four-site intradermal booster regimen with use of one-fifth of the diluent volume of cell-culture rabies vaccine on day 0 is associated with a significantly higher antibody response than is the conventional booster regimen for subsequent postexposure rabies treatment of individuals who have received preexposure rabies vaccination with cell-culture rabies vaccine 1 year previously. PMID- 10452643 TI - Streptococcal meningitis in adult patients: current epidemiology and clinical spectrum. AB - Streptococci other than Streptococcus pneumoniae are a rare cause of bacterial meningitis in adults. We report 29 cases of streptococcal meningitis (1977-1997). The patients comprised 19 men and 10 women, with a mean age +/- standard deviation of 47 +/- 18 years. Nine cases were secondary to neurosurgical procedures, seven to brain abscess, five to cerebrospinal fluid pericranial fistula, and three to endocarditis. Causative microorganisms included the following: viridans group streptococci, 20 cases; anaerobic streptococci, 3; Streptococcus agalactiae, 3; Streptococcus bovis, 2; and Streptococcus pyogenes, 1. Four Streptococcus mitis strains showed decreased susceptibility to penicillin (MIC, 0.5-2 microg/mL). Five patients (17%) died. The infection is increasing in the hospital setting. Streptococci resistant to penicillin should be considered in the empirical treatment of nosocomial meningitis. In cases of community acquired infection, anaerobic streptococci or streptococci of the Streptococcus milleri group should alert the clinician to the presence of an undiagnosed brain abscess, whereas oral streptococci of the viridans group suggest the diagnosis of bacterial endocarditis. PMID- 10452644 TI - Treatment of human immunodeficiency virus 1-infected infants and children with the protease inhibitor nelfinavir mesylate. AB - An open-label study was conducted of nelfinavir mesylate, given with reverse transcriptase inhibitors to human immunodeficiency virus 1 (HIV-1)-infected infants and children 3 months to 13 years of age. Doses of nelfinavir mesylate of 20-30 mg/kg yielded drug exposures comparable to those seen in adults. The drug was well tolerated; mild diarrhea was the primary toxic effect observed. Seventy one percent (39) of the 55 evaluable subjects had an initial decrease in plasma HIV-1 RNA, of at least 0.7 log10 copies/mL; suppression of plasma HIV-1 RNA levels to < 400 copies/mL was observed in 15. Children who began taking at least one new reverse transcriptase inhibitor near the time when nelfinavir mesylate was started, and those with a > or = 24% proportion of CD4 lymphocytes, had a greater chance of achieving and maintaining a decline in plasma HIV-1 RNA to < 400 copies/mL. Suppression of viremia was achieved in children as young as 3 months of age. PMID- 10452645 TI - Determinants of vancomycin use in adult intensive care units in 41 United States hospitals. AB - We analyzed data from a prospective observational cohort study that included 108 adult intensive care units (ICUs) in 41 United States hospitals. Use of vancomycin (defined daily doses per 1,000 patient-days), nosocomial infection rates, and proportion of all Staphylococcus aureus isolates resistant to methicillin (MRSA rate) were recorded from January 1996 through November 1997. The median rate of vancomycin use was lowest in coronary care ICUs and highest in general surgical ICUs. Prior approval before use of vancomycin was required in only 26 (24%) of the 108 ICUs. In a multivariate linear regression model, rates of MRSA, central line-associated bloodstream infection, and the type of ICU were independent predictors of vancomycin use. None of the vancomycin control practices was associated with lower rates of vancomycin use; however, it is important to recognize that this database was not designed to measure rates of inappropriate use. Vancomycin use is heavily determined by rates of endemic MRSA and central line-associated bloodstream infection. Efforts to reduce these rates through infection control activities should be included in hospitals' efforts to reduce vancomycin use. PMID- 10452646 TI - Managing antibiotic use--impact of infection control. PMID- 10452647 TI - Epidemiology and clinical outcomes of patients with multiresistant Pseudomonas aeruginosa. AB - We conducted a case-series study of multiresistant Pseudomonas aeruginosa in patients who did not have cystic fibrosis. Patient characteristics, antibiotic exposures, time course of emergence of resistance, and clinical outcomes were examined. Twenty-two patients were identified from whom P. aeruginosa resistant to ciprofloxacin, imipenem, ceftazidime, and piperacillin was isolated. Nineteen (86%) had clinical infection. Patients received prolonged courses of antipseudomonal antibiotics before isolation of multiresistant P. aeruginosa. Nine of 11 patients with soft-tissue infection exhibited resolution of clinical infection but usually required surgical removal of infected tissue with or without revascularization. Overall, three patients died. In two instances in which multiple isolates with different susceptibility profiles from the same patient were available, pulsed-field gel electrophoresis profiles of serial isolates were indistinguishable or closely related. This study illustrates that multiresistant P. aeruginosa emerges in a stepwise manner after exposure to antipseudomonal antibiotics and results in adverse outcomes. PMID- 10452648 TI - Successful treatment of ceftazidime-resistant Klebsiella pneumoniae ventriculitis with intravenous meropenem and intraventricular polymyxin B: case report and review. AB - Increasing prevalence of multidrug-resistant gram-negative organisms has led to a rise in clinically significant infections with these organisms and an increasing therapeutic dilemma. We present a case of a neurosurgical patient who developed ventriculoperitoneal shunt-associated ventriculitis due to ceftazidime-resistant Klebsiella pneumoniae susceptible to cefepime, imipenem, meropenem, and polymyxin B only. Successful management was accomplished by removal of the shunt and therapy with systemic meropenem and intraventricular polymyxin B. Rapid cerebrospinal fluid (CSF) sterilization occurred, with CSF bactericidal titers of 1:32 to 1:128. Polymyxin B should be considered as adjunctive therapy for life threatening multidrug-resistant gram-negative infections. Prior literature on use of intrathecal polymyxin B in therapy for meningitis supports its potential efficacy. PMID- 10452649 TI - Clinical significance of Candida species isolated from cerebrospinal fluid following neurosurgery. AB - Twenty-one patients for whom adequate clinical data were available were identified in a retrospective review of cases of Candida species isolated from cerebrospinal fluid (CSF) following neurosurgery; 86% had indwelling cerebrospinal devices (shunts). Candida species were isolated from multiple CSF samples from 10 patients; CSF samples from seven of 10 were initially drawn through indwelling devices and those from nine of 10 were obtained by subsequent lumbar punctures. All of these patients were treated with antifungals, although therapy was delayed in 50% of cases until the second positive culture was reported. In 11 cases, Candida was the only isolate recovered from CSF samples drawn through indwelling devices; cultures of subsequent CSF samples obtained by lumbar puncture were negative in 10 of 11 cases. Only two patients for whom a single culture was positive for Candida species were treated with antifungals (both of whom were symptomatic), and none of the untreated patients died of infection. The clinical significance of a single positive CSF sample drawn through an indwelling device is difficult to assess, and a definitive diagnosis may require repeated cultures of CSF samples obtained by lumbar puncture. PMID- 10452650 TI - Infusion of lipoproteins into volunteers enhances the growth of Candida albicans. AB - Infusion of reconstituted high-density lipoproteins (rHDL) is being studied in clinical trials as an adjunctive therapy for gram-negative sepsis. Since no data are available on its possible effects in systemic candidiasis, we investigated the effect of rHDL infusion into volunteers on the growth of Candida albicans. C. albicans growth was 10- to 100-fold higher in the plasma of volunteers infused with 80 or 100 mg/kg rHDL than in plasma collected before infusion; administration of 60 mg/kg rHDL had marginal effects. In vitro, the isolated lipoprotein subfractions had a growth-promoting effect on C. albicans. These data suggest potential adverse effects of rHDL if infused into patients with systemic candidiasis. Thus, rHDL infusion into patients with sepsis caused by an unknown microorganism may be contraindicated. PMID- 10452651 TI - Progressive multifocal leukoencephalopathy in patients with AIDS receiving highly active antiretroviral therapy. AB - Recent reports suggest that human immunodeficiency virus (HIV)-associated progressive multifocal leukoencephalopathy (PML) may improve with highly active antiretroviral therapy (HAART). We observed three patients who developed PML while receiving HAART. All patients received HAART for 4-11 months and had low plasma levels of HIV-1 RNA before the onset of symptoms of PML. Antiretroviral therapy was changed in two patients, and their plasma HIV-1 RNA levels declined significantly. Despite this virologic response, PML did not improve in these patients. The third patient's HIV-1 RNA level became undetectable while he was receiving HAART, and his symptoms of PML improved after the addition of interferon alpha. Our observations suggest that PML can develop in patients who have shown clinical response to HAART. Furthermore, PML may not improve despite an adequate virologic response to HAART. Definitive therapy is still needed for PML. PMID- 10452652 TI - Bartonella henselae associated with Parinaud's oculoglandular syndrome. AB - Bartonella henselae was recovered from the conjunctival scraping of a 38-year-old woman who presented with a 2-week history of tender preauricular lymphadenopathy and a 1-day history of a red left eye. Dry adherent colonies were observed on agar plates at 21 days of incubation, and the isolate was identified through conventional and molecular tests. Polymerase chain reaction (PCR) amplification of a specific region of the 16S rRNA gene and confirmation by a separate PCR reaction with hybridization of the product with a B. henselae-specific probe confirmed the isolate as B. henselae. This is the first reported isolation of the causative agent of cat scratch disease from ocular tissue in a patient with Parinaud's oculoglandular syndrome. PMID- 10452653 TI - Liver abscess due to a Lactobacillus rhamnosus strain indistinguishable from L. rhamnosus strain GG. PMID- 10452654 TI - Pneumococcal pneumonia in adults treated at University of Kentucky Medical Center, 1995-1998: implications of pathogen resistance. PMID- 10452655 TI - Sepsis due to Streptococcus pneumoniae in a patient with alcoholism who received pneumococcal vaccine. PMID- 10452656 TI - Pneumocystis carinii pneumonia mimicking Wegener's granulomatosis. PMID- 10452657 TI - Correlation between in vitro quinolone susceptibility of Neisseria gonorrhoeae and outcome of treatment of gonococcal urethritis with single-dose ofloxacin. PMID- 10452658 TI - Noncardiogenic pulmonary edema during vivax malaria. PMID- 10452659 TI - Diagnostic polymerase chain reaction for donovanosis. PMID- 10452660 TI - Paecilomyces lilacinus infection in a heart transplant recipient and successful treatment with terbinafine. PMID- 10452661 TI - Local spread of molluscum contagiosum by electrolysis. PMID- 10452662 TI - Guillain-Barre syndrome as first manifestation of typhoid fever. PMID- 10452663 TI - Bacteremia due to Capnocytophaga species in patients with neutropenia: high frequency of beta-lactamase-producing strains. PMID- 10452664 TI - Persistence of Salmonella species in cerebrospinal fluid of patients with meningitis following ceftriaxone therapy. PMID- 10452665 TI - Bacteremia due to motile Enterococcus species: clinical features and outcomes. PMID- 10452666 TI - Worsening of endogenous Candida albicans endophthalmitis during therapy with intravenous lipid complex amphotericin B. PMID- 10452667 TI - A DNA-based probe for differentiation of Giardia lamblia group A and B isolates from northern India. PMID- 10452668 TI - Two episodes of acute renal failure, rhabdomyolysis, and severe hepatitis in an AIDS patient successively treated with ritonavir and indinavir. PMID- 10452669 TI - Serratia marcescens cellulitis following an iguana bite. PMID- 10452670 TI - Vivax malaria complicated by adult respiratory distress syndrome. PMID- 10452671 TI - Thyroid abscess caused by Mycobacterium chelonae. PMID- 10452672 TI - Prospective follow-up of 67 indinavir-experienced human immunodeficiency virus infected and AIDS patients treated with the ritonavir/saquinavir combination. PMID- 10452673 TI - Is a proton pump inhibitor necessary for the treatment of lower-grade reflux esophagitis? AB - The efficacy of histamine H2 receptor antagonist (H2RA) and proton pump inhibitor (PPI) therapies in healing reflux esophagitis was compared in a prospective randomized case-control comparative study. A total of 71 patients with grade A to D esophagitis (Los Angeles classification) were given either famotidine 20 mg twice a day (Fam; n = 35) or omeprazole 20 mg once daily (Ome; n = 36) for 8 weeks. Endoscopy was performed to assess healing in 57 patients. Healed patients were followed-up without H2RA or PPI therapy for 3 months. At the end of follow up, endoscopy was able to be performed in 33 patients. Healing rates for patients in the Fam and Ome groups were 58.6% (17/OFF and 97.4% (27/28), respectively (P < 0.001), and when limited to grade A to B, healing rates were 60.9% (14/23) and 100% (25/25), respectively (P < 0.001). Concerning Helicobacter pylori infection, healing rates for the Fam and Ome groups in H. pylori (+) patients were 90.0% (9/10) and 90.9% (10/11), respectively (P = 1.00). Remission rates in the Fam and Ome groups were 45.0% (91/2)) and 33.3% (6/18), respectively (P > 0.4). In regard to alcohol drinking, remission rates of daily and social drinkers were 7.7% and 42.4%, respectively (P < 0.03). Thus, PPI should be the drug of choice even for healing lower-grade reflux esophagitis, especially in H. pylori (-) patients. Treatment with H2RA may be an alternative choice in H. pylori (+) patients. After healing, most patients cannot sustain remission without maintenance therapy. PMID- 10452674 TI - Proportion of reflux esophagitis in 6010 Japanese adults: prospective evaluation by endoscopy. AB - Compared with findings in Western countries, the prevalence of reflux esophagitis in Oriental countries is estimated to be low. In this prospective study, we aimed to examine the proportion of reflux esophagitis in Japanese adults, as evaluated by endoscopy. Endoscopists were prospectively directed to grade esophageal mucosal breaks with esophagitis according to the Los Angeles Classification of Esophagitis in all subjects that underwent endoscopic examination. In total, 6010 subjects underwent endoscopic examination for evaluation of esophagitis grading from December 1996 to February 1998. The subjects included 4394 outpatients who were not receiving medication for gastrointestinal disease and 1616 subjects who visited the hospital for routine physical examinations. The overall proportion of esophagitis was 16.3%. Most of the subjects with esophagitis were classified as having grade A or B (proportion of grades A and B, 9.6% and 4.6%, respectively). The age-related proportion of esophagitis and of severe esophagitis (i.e., grades C and D) increased in females aged over 70 and in males aged over 80. Increased body mass index (partly due to decreased height caused by osteoporosis), and/or hiatal herniation, were related to the proportion of esophagitis in females aged over 70. These data indicated that reflux esophagitis is a common disease in Japan. However, severe esophagitis (grades C and D) is not common. PMID- 10452675 TI - Effects of aging and gastric lipolysis on gastric emptying of lipid in liquid meal. AB - Lipid delays gastric emptying, and aging is associated with changes in gastric motor function and transit. However, little is known about the effect of lipid on gastric emptying time in the elderly. To determine the effect of aging on lipid gastric emptying, we used electrical impedance tomography (EIT) to study gastric emptying of liquid meals with or without lipid in five young (23.0 +/- 0.6 years, mean +/- SEM) and six elderly (73.3 +/- 1.6 years) healthy male volunteers. These subjects drank 400 ml of non-lipid soup (triglycerides, 0 g) or lipid soup (triglycerides, 24.6g) in liquid test meals. To study the effect of lipolysis in the stomach, a liquid test meal containing 240mg of lipase in the lipid soup was also administered. Plasma cholecystokinin (CCK) concentration was measured by specific radioimmunoassay before and 30 min after the ingestion of a test meal. The gastric emptying time of the lipid soup was longer in the elderly than in the young subjects, and the time was significantly longer for lipid soup than for nonlipid soup (P < 0.05) in both the young and elderly subjects. Gastric emptying time for non-lipid soup was not significantly different between the elderly and young subjects. The administration of lipase shortened the gastric emptying time for lipid in both the elderly and the young subjects. Basal CCK concentration was significantly higher in the elderly than in the young subjects. However, there was no relationship between gastric emptying time and plasma CCK concentration after the ingestion of a test meal in the subjects overall. In conclusion, the delaying effect of lipid on gastric emptying is increased in the elderly, and the administration of lipase accelerates the emptying of lipid from the stomach. PMID- 10452676 TI - Development of poorly differentiated adenocarcinoma and carcinoid due to long term Helicobacter pylori colonization in Mongolian gerbils. AB - A Mongolian gerbil model was used to clarify whether long-term colonization by Helicobacter pylori is an important risk factor for the development of gastric cancer. Fifty-nine gerbils (3 controls and 56 gerbils inoculated with H. pylori) were killed at various times (average, 23 months) more than 12 months after H. pylori inoculation. In the H. pylori-inoculated group, poorly differentiated adenocarcinoma was observed in the pylorus of 1 gerbil, and carcinoid was observed in the fundus of the stomach in 18 gerbils. No lesions were found in the stomachs of the 3 control gerbils. The results imply that long-term colonization by H. pylori is an important risk factor for the development of gastric adenocarcinoma and carcinoid. PMID- 10452677 TI - Helicobacter pylori-negative gastric and duodenal ulcers. AB - It is unclear whether Helicobacter pylori infection is essential to the development of peptic ulcers. In this study, we examined the rates of H. pylori negativity among patients with peptic ulcers. We also attempted to clarify the characteristics of H. pylori-negative peptic ulcers to throw light on the pathogenesis of peptic ulcers. The study included 215 consecutive patients with gastric ulcers (GUs) and 120 consecutive patients with duodenal ulcers (DUs). After routine endoscopic examination and phenol red dye endoscopy, forceps biopsies were performed for culture, histology, and the rapid urease test. A patient was considered H. pylori-negative when the serum anti-H. pylori IgG and the three tests on biopsied specimens were all negative. H. pylori-negative rates were 3.2% in the patients with GUs and 1.7% in the patients with DUs. Lack of atrophy of the gastric mucosa was significantly more common in the H. pylori negative patients with GUs. A history of ulcer disease was less common and antral ulcers were more common in H. pylori-negative GU patients, but not significantly so. As the urea breath test had not been performed, the possibility of a false negative result cannot be completely ruled out, but we believe that the H. pylori negative rate in our study is more reliable than these rates in previous reports, because we visualized H. pylori distribution by phenol red dye endoscopy to avoid false-negative results in biopsies, and we used both biopsy and serum anti-H. pylori IgG findings to establish an H. pylori-negative diagnosis. Since H. pylori negative peptic ulcers certainly exist, H. pylori infection is thought not to be essential to the development of peptic ulcers. There were few differences between the characteristics of H. pylori-negative and H. pylori-positive peptic ulcers in our study. A large-scale study is required to clarify the characteristics of H. pylori-negative peptic ulcers. PMID- 10452678 TI - Relationship between Helicobacter pylori infection and histologic features of gastritis in biopsy specimens in gastroduodenal diseases, including evaluation of diagnosis by polymerase chain reaction assay. AB - We investigated the relationship between Helicobacter pylori infection and the histologic features of gastritis in gastroduodenal disease, and evaluated the diagnostic usefulness of the polymerase chain reaction (PCR) assay for the detection of H. pylori before and after eradication therapy. Endoscopic biopsy specimens from 81 patients with gastroduodenal disease were examined for the presence of H. pylori by culture and histologic examination. Histologic features of gastritis were classified according to the updated Sydney System, and results of the PCR assay were compared with those of histologic examination, using histologic scores. The density of H. pylori was significantly correlated with polymorphonuclear neutrophil activity and chronic inflammation. These findings suggest that the grades of infiltration of polymorphonuclear neutrophil cells and chronic inflammatory cells correspond to the density of H. pylori infection assessed by the updated Sydney System. Patients with positive results on PCR assay and negative results on histologic examination may have a low density of H. pylori because of severe atrophy in the gastric mucosa. Differences in results for the PCR assay and histologic examination were found in 2 of 12 patients in the detection of H. pylori after eradication therapy. According to the results of the PCR assay and histologic features before and after eradication, the gastric tissue-based PCR assay for H. pylori after eradication may be too sensitive to judge successful eradication of H. pylori. PMID- 10452679 TI - Superficial blood flow, blood volume, and blood velocity in colorectal tubular adenomas and adenocarcinomas. AB - To clarify differences in the vasculature of benign and malignant colorectal tumors, we compared blood flow (which has been examined in previous studies) as well as blood volume and blood velocity in two types of human colorectal tumors. We measured blood flow and blood volume with an endoscopic laser-Doppler flowmeter in the lesions and the normal mucosa near the lesions of 79 tubular adenomas with mild dysplasia and 38 adenocarcinomas. Blood velocity was defined as blood flow per blood volume. The ratio of each value in the lesions to that in the neighboring normal mucosa was defined as the relative value, and the mean relative values in the two groups were compared. The relative blood flow was 1.89 +/- 0.64 in the tubular adenomas and 1.49 +/- 0.66 in the adenocarcinomas (P < 0.005), and the relative blood volume was 1.10 +/- 0.14 and 0.90 +/- 0.13, respectively (P < 0.001). The relative blood velocity was approximately 1.7 in both groups (NS). This study demonstrated that the superficial blood flow in adenocarcinomas is increased due to increased blood velocity, despite blood volume being decreased. PMID- 10452680 TI - Estimation of glycogen levels in human colorectal cancer tissue: relationship with cell cycle and tumor outgrowth. AB - In this study, we quantitatively measured glycogen levels in tissue samples obtained from tumors, regions adjacent to tumor, and regions of normal colorectum to determine whether the levels were related to cell cycle and cancer growth. Glycogen levels were analyzed in relation to histopathological factors, (tumor size and stage of disease) and cell cycle progression. The glycogen level was found to be highest in the cancer tissue, lower in normal tissue, and lowest in the adjacent tissue. The difference in glycogen level between the cancer tissue and the other two regions was significant (P < 0.05). There was a negative correlation between glycogen level and tumor size, but it was not significant. The level of glycogen in cancer tissues decreased as the stage of the disease progressed, but a significant difference was not found between stages. There was a negative correlation between the glycogen level and the proliferation index. There was a positive correlation between the glycogen level and the proportion of cancer cells in G1 phase, while there was a negative correlation with S and G2M phases. Glycogen levels were highest in cancers with a high proportion of cells in G1, and decreased with progression to S phase. It may be that glycogen is utilized in the progression to S phase, and the cancer tissues are supplied with glycogen from the tumors themselves as well as their adjacent tissues. Cancer growth may be inhibited by artificial control of the glycogen level in the G1 phase of cancer cells. PMID- 10452681 TI - Suppression of hepatitis C virus by hepatitis B virus in coinfected patients at the National University Hospital of Singapore. AB - This study was carried out to compare hepatitis B virus (HBV) and hepatitis C virus (HCV) replicative states and to determine possible interference between HBV and HCV. One thousand and sixty-one consecutive patients seen at The Gastroenterology Division of the National University Hospital of Singapore between 1988 and 1995 were screened for HBV and HCV serological markers. Anti-HCV was tested using a second-generation enzyme-linked immunosorbent assay (ELISA) method. HCV-RNA was detected by a reverse-transcription polymerase chain reaction method (RT-PCR). Data were analyzed by either paired t-test or chi2 test. Two hundred and twenty-four patients were infected with HBV alone, while 117 patients were infected with HCV only. Thirty-one patients were coinfected with HBV and HCV. HCV-RNA was detected in 104 of the 117 HCV patients (88.9%), and in 12 of 29 coinfected patients (41.4%). Serum anti-HCV levels in the coinfected patients were lower than those in the HCV-infected patients. A significant difference for anti-HCV reactivity and HCV-RNA positivity was observed between HCV-infected patients and coinfected patients (P < 0.01). In contrast, HBV-DNA and hepatitis B envelope antigen (HBeAg) positive rates were similar in HBV carriers and patients coinfected with HBV and HCV. These results show a possible interaction between HBV and HCV life cycles, and suggest that HCV replication may be negatively affected by HBV. PMID- 10452682 TI - Association of cyclin D1 expression with factors correlated with tumor progression in human hepatocellular carcinoma. AB - The amplification and/or rearrangement of the cyclin D1 gene, a positive regulatory element of the G1 to S phase of the cell cycle, has been reported in various human tumors, suggesting an oncogenic role of this gene. In this study, we investigated the expression of cyclin D1 in the formalin-fixed and paraffin embedded human hepatocellular carcinoma (HCC) tissues of 25 patients, using monoclonal antibody 5D4 raised against cyclin D1. Two distinct patterns of staining were observed in HCC cells, nuclear and cytoplasmic. The nuclear staining pattern of cyclin D1 was detected in the tissues of only 2 of the 25 HCC patients (8%) examined and no particular clinicopathological characteristics were found in these patients. In contrast, the cytoplasmic staining pattern, without nuclear staining, was detected in 8 of the 25 patients with HCC (32%). A significant correlation was found between the expression of cytoplasmic cyclin D1 and patients with tumor thrombus in the portal vein (Vp), as well as those with intrahepatic metastasis (IM). These results indicate that the cytoplasmic cyclin D1 expression appears to be related to the prognosis of HCC. The Ag nucleolar organizer regions (NORs) counts in cyclin D1-positive and -negative patients were not significantly different, suggesting that immunostaining for cyclin D1 has the potential to be a unique prognostic marker in human HCC. Simultaneous immunohistochemical study with p53 antibody in the same series of HCC revealed that 88% of the patients positive for cyclin D1 also expressed p53 and that in 91% of the patients negative for p53, cyclin D1 was not expressed. These results suggest that cyclin D1 is expressed later than the alteration of p53 in the progression of human HCC. PMID- 10452683 TI - Little contribution of GB virus C to the development of primary hepatocellular carcinoma in Japan. AB - RNA of GB virus C was searched for in sera from 109 patients with primary hepatocellular carcinoma. It was detected in 11 patients (10%), and was more frequent than in 3 of 342 blood donors (0.9%) (P < 0.001). The 11 patients included 4 of 29 patients (14%) with hepatitis B surface antigen and 7 of 74 patients (9%) with antibody to hepatitis C virus. RNA of GB virus C was not detected in any of 4 patients without hepatitis B surface antigen or antibody to hepatitis C virus. These results suggest that GB virus C may contribute little to the development of primary hepatocellular carcinoma in Japan. PMID- 10452684 TI - Hepatocyte growth factor stimulates cell growth and enhances the expression of transforming growth factor alpha mRNA in AsPC-1 human pancreatic cancer cells. AB - We investigated the effects of hepatocyte growth factor (HGF) and transforming growth factor alpha (TGF alpha) on cell growth in four human pancreatic cancer cell lines. Changes in the expression of mRNAs of HGF, c-met, TGF alpha, and epidermal growth factor receptor (EGFR) by treatment with HGF and TGF alpha were observed. Cell growth with growth factors was assessed with the MTT assay and compared with basal growth without growth factors. Although HGF stimulated cell growth in AsPC-1, COLO-357, and T3M4 cells, Panc-1 cells showed no response to HGF. TGF alpha stimulated the growth of all the above cells. The expression of c met mRNA under nonstimulated conditions was detected with Northern blotting in all cells. Treatment with HGF slightly enhanced the expression of c-met mRNA only in COLO-357 cells. The intensity of EGFR expression was consistent, and HGF mRNA was not detected during induction experiments in any cell type. Concomitant treatment with HGF and TGF alpha exerted an effect that was additive or less on the growth of all cells. Expression of TGF alpha was enhanced by HGF treatment only in AsPC-1 cells. These results suggested that HGF and TGF alpha stimulated cell growth through a final common pathway of signal transduction. PMID- 10452685 TI - Enzyme-linked immunosorbent assay to detect hepatitis C virus serological groups 1 to 6. AB - By conventional serological grouping methods, it is possible to determine hepatitis C virus (HCV) serological groups for genotypes 1a, and 1b, and genotypes 2a, and 2b, but not for other genotypes, i.e., 3a, 3b, 4a, 5a, and 6a. In this study, we attempted to serologically group HCV with the Murex HCV serotyping 1 to 6 assay (Murex Diagnostics, Kent, UK), using an enzyme-linked immunosorbent assay (ELISA) based on genotype-specific peptides from the NS4 region. The subjects of this study were 365 patients infected with HCV of genotype 1a, 1b, 2a, 2b, 3a, or 3b. The sensitivity of the assay was 100% in patients with genotype 1a, 82.7% in those with 1b, 68.5% in those with 2a, 84.2% in those with 2b, 50.0% in those with 3a, and 76.5% in those with genotype 3b. The overall sensitivity was 78.4%. The specificity of the assay was 100% in the subjects with genotype 1a, 98.8% in those with 1b, 98.4% in those with 2a, 96.9% in those with 2b, 100% in those with 3a, and 100% in those with genotype 3b. The overall specificity was 98.6%. The concordance of the assay was 100% in subjects with genotype la, 81.7% in those with 1b, 67.4% in those with 2a, 81.6% in those with 2b, 50.0% in those with 3a, and 76.5% in those with genotype 3b. The overall concordance was 77.5%. We believe it would be better to serotype with the Murex HCV serotyping 1 to 6 assay, if other than serological group (Gr) 1 or Gr 2 is suspected in particular ethnic groups or in subjects with an indeterminate result with the Immucheck HCV Gr assay (Kokusai, Kobe, Japan), assuming that the genotype must be other than 1a, 1b, 2a, or 2b. PMID- 10452686 TI - Esophageal carcinoma with high serum parathyroid hormone-related protein (PTHrP) level. AB - We report two patients with esophageal carcinoma with high levels of serum parathyroid hormone-related protein (PTHrP). Patient 1 was a 66-year-old man in whom the serum calcium level was also high, and patient 2 was an 81-year-old woman. The serum PTHrP level was 411 pM (normal range, 13.8-55.3pM) in patient 1 and 94.5 pM in patient 2 (in whom the serum granulocyte colony-stimulating factor level was also high). We demonstrated PTHrP immunohistologically in esophageal carcinoma cells in both patients. After admission, patient 1 died of pneumonia on the 17th day of hospitalization (the 48th day after he had had an episode of frequent hiccuping) and patient 2 died of acute circulatory failure on the 12th day of hospitalization (the 25th day after she had vomited after a meal). Neither of these patients died of cancer. Pneumonia in patient 1 was believed to be due to weakened body defenses, while the acute circulatory failure in patient 2 was due to emaciation. Since esophageal carcinoma with humoral hypercalcemia of malignancy and leukocytosis is characterized by rapid progression and metastasis, early diagnosis and treatment are mandatory. PMID- 10452687 TI - A patient with rectal cancer associated with ulcerative colitis in whom endoscopic ultrasonography was useful for diagnosis. AB - Endoscopic ultrasonography (EUS) was helpful for the diagnosis of rectal cancer associated with ulcerative colitis. The patient was a 38-year-old Japanese man with a 19-year history of relapsing-remitting type ulcerative colitis involving the entire colon. Routine colonoscopy revealed multiple polypoid prominences in the upper portion of the rectum. EUS revealed a hypoechoic mass in the submucosa beneath and around the polypoid lesion on the most oral side. Signet ring cells were found in a biopsy specimen from this lesion. Subtotal colectomy was performed. A depressed lesion was observed around the prominence on the most oral side; histologically, this lesion was poorly differentiated mucinous and signet ring cell carcinoma extending into the subserosa. The polypoid lesion on the most anal side was well differentiated adenocarcinoma, which was limited to the mucosa. Our findings suggest that EUS is helpful for detecting invasive cancer associated with ulcerative colitis. PMID- 10452688 TI - Cystic lymphangiomas of the colon. AB - We report a patient with cystic lymphangiomas diagnosed by endoscopic ultrasonography and resected by partial polypectomy. A 42-year-old woman consulted a nearby physician because of a positive fecal occult blood test. Barium enema and colonoscopy revealed the presence of abnormalities. On March 11, 1997, she was admitted to our department for further evaluation and treatment. A barium enema examination revealed two protruding lesions in the transverse colon. Colonoscopy showed a teardrop-type mass in the left side of the transverse colon. The mass was cushion-sign positive, and its shape readily changed on respiration and with changes in body position. Another superficial smooth mass was found in the right side of the transverse colon. Ultrasonography of the colon confirmed the presence of a submucosal mass showing a cyst-like pattern. Cystic lymphangiomas were diagnosed and resected endoscopically. Histopathological examination revealed markedly dilated ducts consisting of a single layer of endothelial cells in the submucosa of the colon. The diagnosis was cystic lymphangioma. PMID- 10452689 TI - Massive rectal bleeding due to ileal tuberculosis. AB - A patient with massive rectal bleeding due to ileal tuberculosis is reported. Technetium-99m labelled red blood cell scintigraphy indicated hemorrhage from the ileum, and laparotomy was then carried out. A 70-cm segment of ileum containing ulcers and erosions was resected, and epitheloid granuloma with Langhans-type giant cell was found in the resected specimen. Massive rectal bleeding is considered a rare presenting symptom of intestinal tuberculosis. Intestinal tuberculosis, including small intestinal tuberculosis, although uncommon, should be taken into consideration as a cause of rectal bleeding. PMID- 10452690 TI - Hypertension as a paraneoplastic syndrome in hepatocellular carcinoma. AB - We report a 66-year-old man with hepatocellular carcinoma who was positive for hepatitis B surface antigen, and was hospitalized because of hypoglycemia and hypertension. His plasma renin activity was normal (2.3 ng/ml per h), but concentrations of angiotensin I (>2500 pg/ml) and II (86 pg/ml) were high. Increased angiotensin I level at sites proximal and distal from the confluence of the hepatic vein and the inferior vena cava indicated that the hypertension was provoked by overproduction of angiotensin I from the hepatocellular carcinoma. Previous reports of patients with hepatocellular carcinoma with hypertension due to abnormality of renin-angiotensin system are reviewed. PMID- 10452691 TI - Papillary adenoma of the distal common bile duct. AB - A 73-year-old man with a papillary adenoma located in the distal common bile duct is reported. He underwent pylorus-preserving pancreatoduodenectomy. The lesion in the common bile duct featured papillary proliferation of the epithelium and fibrous elements with diffuse infiltration by inflammatory cells. Positive staining for MIB-1 (Ki-67) and p53 was identified in the nuclei of the proliferative epithelium. These findings suggested the malignant potential of this lesion. Further progress in imaging diagnostic techniques should increase the frequency with which such lesions are discovered. Even now, if mural irregularities and defects are found in the extrahepatic biliary system, especially the distal common bile duct, the possibility of such borderline biliary adenoma should be taken into consideration when making a diagnosis. PMID- 10452692 TI - Optimum management of mild esophagitis in Japan. PMID- 10452693 TI - Interaction among fat, lipase, CCK, and gastric emptying. PMID- 10452694 TI - One more new gastric disease induced by Helicobacter pylori infection, enterochromaffin-like (ECL) cell carcinoid tumor. PMID- 10452695 TI - Does cytoplasmic cyclin D1 regulate the cell cycle directly? PMID- 10452696 TI - Hepatocyte growth factor and pancreatic cancer cells. PMID- 10452697 TI - Decisional balance regarding substance use among persons with schizophrenia. AB - State-of-the-science treatment of substance abuse relies on decisional balance activities (weighing pros and cons of continued substance use) to enhance motivation for change. Few data are available regarding the feasibility of these activities among persons dually diagnosed with schizophrenia and substance use disorder. To address this lacuna in the literature, we completed focus groups with 21 participants, all of whom had a schizophrenia-spectrum diagnosis and lifetime substance abuse or dependence. These key informants discussed the pros and cons of substance use as well as the pros and cons of quitting in response to a structured group interview. Our qualitative data indicate that persons living with schizophrenia can generate rich and diverse decisional balance information. We describe salient themes, contrast complementary perspectives (i.e., the pros of using and cons of quitting), and suggest treatment implications based on these findings. PMID- 10452698 TI - Transformational and transactional leadership skills for mental health teams. AB - Many treatments for persons with severe mental illness are provided by mental health teams. Team members work better when led by effective leaders. Research conducted by organizational psychologists, and validated on mental health teams, have identified a variety of skills that are useful for these leaders. Bass (1990, 1997) identified two sets of especially important skills related to transformational and transactional leadership. Leaders using transformational skills help team members to view their work from more elevated perspectives and develop innovative ways to deal with work-related problems. Skills related to transformational leadership promote inspiration, intellectual stimulation, individual consideration, participative decision making, and elective delegation. Mental health and rehabilitation teams must not only develop creative and innovative programs, they must maintain them over time as a series of leader-team member transactions. Transactional leadership skills include goal-setting, feedback, and reinforcement strategies which help team members maintain effective programs. PMID- 10452699 TI - The impact of a Community Mental Health Center on psychiatric hospitalizations in two Athens areas. AB - This paper explores the impact of a Community Mental Health Center intervention activities on the inpatient psychiatric morbidity of two areas served by this center. Athens University established this Center, the first of its kind in the Greater Athens area. A comparison among the utilization rates of inpatient psychiatric services by community residents during the years 1979, 1985, 1991 and 1995 is made. The results of the 1995 survey show a significant reduction in the number and days of hospitalization and a remarkable cut in compulsory admissions when compared with those in 1979. It is concluded that medication monitoring, outreach, domiciliary care for patients in crisis, and day care are effective and robust principles of mental health services. PMID- 10452700 TI - Case management models for persons who are homeless and mentally ill: the ACCESS demonstration project. Access to Community Care and Effective Services and Supports. AB - Persons who are homeless and mentally ill present unique challenges to service providers and human service systems. In vivo case management approaches such as assertive community treatment (ACT) have shown promise in engaging this population. This paper explores case management models employed within the ACCESS program, a five year, 18-site demonstration program enriching services for homeless persons with serious mental illness. We describe the implementation of case management with ACCESS programs and determine the extent of variation across sites using a measure of fidelity to ACT. While programs reported using four models, much similarity was found among programs on multiple dimensions. PMID- 10452701 TI - The mental health consumer movement: implications for rural practice. AB - Developing consumer-oriented programs for rural areas presents a major challenge for practitioners and policy makers. The mental health consumer movement, a successful urban creation, has yet to fully impact rural practice and be of benefit to individuals with severe and persistent mental illness. Rural mental health professionals face unique challenges and opportunities in utilizing rural strengths to foster consumer participation in the design and implementation of service delivery. The authors address the unique barriers facing rural communities and propose a self-help model as a service delivery alternative. PMID- 10452702 TI - A model for multidisciplinary peer review and supervision of behavioral health clinicians. AB - The authors describe a semi-annual program for multidisciplinary peer review and supervision that is in use at two rural adult and child mental health and drug and alcohol service agencies. The program incorporates semiannual chart reviews and face-to-face discussions held with each clinician by a multidisciplinary supervisory group. Several quality improvement issues addressed by this program are described, including improved communication, improved clinical diagnosis, the establishment of service parameters, more appropriate referrals for psychiatric and other medical care, and improved clinician skills. The program represents an inexpensive approach to peer review and supervision that can incorporate university-based consultants, improve quality of care, improve clinicians' skills, and be readily applied by clinical supervisors to most behavioral health settings. PMID- 10452703 TI - Family planning and parenthood needs of women with severe mental illness: clinicians' perspective. AB - Family planning and parenthood are important issues for women with severe and persistent mental illness (SPMI). The role of adult mental health clinicians with regard to these issues has been under investigated. Clinicians treating patients with SPMI at a large community health center completed survey forms on 419 women. Clinicians reported that a large fraction of sexually active women were not thought to be using birth control. Despite this, many clinicians had not discussed birth control with these patients. Clinicians had concerns about childcare in 72% of cases where the patient with SPMI was the primary custodian of a younger child; however, the majority of these families were not receiving child or family services. Further consideration of the role of the adult mental health clinician in addressing issues of family planning and parenting is required. PMID- 10452704 TI - Optical measurement of hematocrit and other biological constituents in renal therapy. AB - Optical sensors have advanced significantly over the past 2 decades leading to today's noninvasive optical measurement capabilities and their widespread applications in renal therapy. These measurements provide significant advantages to the clinician. For example, a given blood constituent can be monitored in real time (continuously, nondestructively), which facilitates the ability to optimally "titrate" the therapy with immediate visual feedback. Optical methods have another intrinsic advantage in that each biologic constituent has its own unique spectral "signature" allowing for simultaneous, multiple, and specific measurements of biologic analytes. Use of this budding spectral technology in renal therapy today provides for increased patient safety (by measuring plasma free hemoglobin, microemboli, clots, oxygen saturation, blood leaks, and hematocrit), measurements of dialysis dose (dialysate urea levels), dry weight (tissue water monitoring), access viability (recirculation, access blood flow), cardiac status (absolute blood volume, cardiac output), and enhanced continuous fluid management (fluid overload, critical blood volume). As microelectronics and signal processing capabilities continue to advance, so will the future of optical diagnosis and treatment. These capabilities translate directly to improved patient quality of life. PMID- 10452705 TI - Cardiac output and central blood volume during hemodialysis: methodology. AB - Cardiovascular disease is the leading cause of mortality in patients whose lives depend on hemodialysis. We developed a method for measuring cardiac output (CO) and central blood volume (CBV) in hemodialyzed patients that may help to elucidate the mechanisms and consequences of cardiac disease in this population. This report describes the technique, focusing on the main sources of error and how they can be prevented. Three principal sources of error were identified: (1) access recirculation (existing or induced during injection); (2) the second pass of the indicator through the cardiopulmonary system, exacerbated by prolonging the duration of intravenous injection; and (3) the transit time of the indicator through the dialysis blood lines. After the algorithms were adjusted to prevent the above errors, the reproducibility of CO and CBV, expressed as the absolute percent deviation from the average of duplicates (3,488 values duplicated within 5 minutes), was 4.3 +/- 3.8% for CO and 4.1 +/- 3.8% for CBV. To determine the clinical value of routine CO and CBV measurements, morbid events (nausea, vomiting, and/or muscle cramps) were prospectively recorded in 73 randomly selected hemodialysis patients. CO and CBV were measured near the beginning and near the end of 98 dialysis sessions during which 28 morbid events were identified. In 10 of these sessions, where morbid events took place within 30 minutes of the measurements, CBV appeared to be a more sensitive indicator of morbid events than CO. We conclude that CO and CBV can be routinely and reliably measured during hemodialysis if precautions are taken to avoid specifically identified sources of error. Preliminary studies suggest that these measurements may have significant prognostic value. PMID- 10452707 TI - Sodium management in dialysis by conductivity. AB - The determination of dialysate sodium concentration is one of the challenges of dialysis prescription, because no accurate information on the predialytic sodium overload is available. Too low dialysate sodium is responsible for intradialytic intolerance symptoms, whereas too high sodium may lead to long-term water sodium overload with cardiovascular hazards (hypertension, left heart failure). We propose here a biofeedback system based on noninvasive repeated measures of ionic dialysance and plasma water conductivity used here as a surrogate of plasma water sodium. This system achieves a stable postdialytic sodium pool and subsequently a dialysate sodium concentration adapted to the inter dialytic sodium load. This new tool in dialysate sodium prescription aims at reducing the morbidity related to patient sodium balance impairment. PMID- 10452706 TI - Hemodynamic monitoring during hemodialysis. AB - Intradialytic monitoring of hemodynamic parameters is an active area of research; future developments in this field will decrease intradialytic morbidity and the mortality of end-stage renal disease patients treated by hemodialysis. Recent investigations have been assisted by the development of devices that can continuously and noninvasively measure hematocrit and plasma protein concentration during the treatment. Intradialytic morbidity, fluid overload, and hypertension in chronic hemodialysis patients have been shown to be associated with either large or small intradialytic decreases in blood or plasma volume that can be routinely measured by these devices. The use of intradialytic changes in blood volume as a feedback control parameter to vary the ultrafiltration rate and dialysate sodium concentration, so called profiling, is now possible, but further research in this area is necessary to show how to optimize the control algorithms. Other, more preliminary studies suggest that monitoring of central blood volume, extracellular volume, and cardiac output during hemodialysis may permit improved hemodynamic stability during treatment and better control of blood pressure. Although optimal application of these techniques and devices remains to be shown, their routine use during maintenance hemodialysis therapy will likely be the standard of care in the near future. PMID- 10452708 TI - Biofeedback systems architecture. AB - The capability for a dialysis machine to use a measurement of the patient's status to automatically tune the dialysis session on-line is commonly addressed by physicians and bioengineers working in the hemodialysis field as "biofeedback." This paper presents the basics of mathematical modeling and control theory normally used in bioengineering, together with some advanced techniques, such as adaptive and multi-input/multi-output control systems. The architectural requirements for implementing biofeedback techniques in renal replacement therapy are then discussed, with due attention paid to the safety aspects, which play a central role in machines hosting such new techniques as well as their therapeutic mission. Finally, the blood volume tracking system, which is aimed at performing the intradialytic water removal, while maintaining a balance inside the body fluids compartments and thus preserving cardiovascular stability, is used as a paradigmatic example of such a class of advanced techniques. The significant results shown by the blood-volume-controlled treatments during a multicenter study focused on its clinical application (30% reduction of intradialysis collapses, 13% reduction of interdialysis symptoms) indicate the technical feasibility and the remarkable benefits of such systems, which get closer to a structurally complete artificial kidney. PMID- 10452709 TI - Urea sensors--a world of possibilities. AB - The arrival on the market of several monitors for the on-line measurement of urea has opened up a new range of possibilities. Already established dose measurements can be performed more easily, and a number of new parameters can be calculated. In this paper, different technologies for urea measurements are first discussed. Devices available on the market are then described according to the technology that is used and the parameters that are calculated. Finally, to show the versatility of this new possibility, a number of additional parameters are discussed that can be determined from dialysate side measurements of urea. PMID- 10452710 TI - Monitoring vascular access flow. AB - Blood flow in peripheral arterio-venous fistulae and grafts as used for hemodialysis access can be derived from measurements of the amount of access recirculation induced by reversing the dialysis blood lines and a knowledge of dialyzer blood flow rates. Furthermore, low or falling access blood flow rates are predictive of access dysfunction from the development of an intraluminal stenosis, which may be reversible, or of access thrombosis. The monitoring of access blood flow rates in hemodialysis populations is therefore suggested. Technologies including ultrasound dilution, hematocrit dilution (or concentration) differential conductivity, and thermodilution used for such monitoring are described. PMID- 10452711 TI - Focus on rehabilitation: teamwork that works. AB - The end-stage renal disease patient has progressed in the past 25 years from the patient who was chosen by a committee to continue life, to a patient who chooses to live well on dialysis. The Vanderbilt Dialysis Clinic (VDC) has developed a collaborative team approach to enable the patient to more effectively achieve this goal. The emphasis has been placed on the patient's "ability, not disability." The collaborative effort of the staff at the VDC has evolved based on the perspective of a team concept. Each discipline or member of the team has a specific goal but functions in a manner that is interdependent of the other team members. The physician, nurses, dietitians, social workers, technicians, and activities director all play an important role in the concept of rehabilitating the whole person. The key to a successful rehabilitation program is to begin with interested patients and staff to serve as a catalyst for others. The Rehabilitation Program at VDC has been based on the principles of continuous quality improvement (CQI) at the unit. The team used the CQI process to brainstorm ideas to implement the exercise program, to overcome barriers to success, and to recognize opportunities to motivate patients and staff. Physicians, nurses, technicians, and dietitians worked together to develop the screening protocols so that the staff would feel comfortable when assessing the patient for exercise. An exercise physiologist from Vanderbilt's Kim Dayani Human Performance Center participated in evaluating the exercise screening information and worked with individual patients and staff members as the program began. Equipment was acquired in the form of used stationary bicycles, smaller ergometer style pedals, hand and ankle weights, and therabands. The exercise program was initiated with only 2 to 4 patients per week participating, after which time it was evaluated and modified. VDC discovered that heightened awareness does indeed impact the level of participation in the facility. The Rehabilitation Team implemented the exercise program in October 1996 with 60% patient participation, yet within 6 months, participation decreased to 25%. One year later in conjunction with a Rehabilitation Fair, interest again peaked to a participation level of 60% in the center and 66% with the home dialysis population. While the improvement in participation may have been due to more quantifiable measurement, it is perhaps more reflective of the emphasis placed on the importance of exercise. Motivators such as an Annual Rehabilitation Fair in which all areas of the Life Options Rehabilitation Advisory Council's "5 E's"--encouragement, education, exercise, employment, and evaluation--are emphasized and have been effective in generating renewed interest among staff and patients in improving overall rehabilitation of patients at the VDC. Continuous motivation of patients and staff through ongoing program implementation, evaluation, and modification based on the CQI process is necessary to maintain everyone's level of interest. PMID- 10452712 TI - Multidisciplinary team renal rehabilitation: interventions and outcomes. AB - Multidisciplinary team renal rehabilitation produces positive patient outcomes by broadening the knowledge base and evening out the work load. Rehabilitation is no longer viewed as an extra program to patient care done by a few people but seen as an integral part of the patients' care and dialysis treatment. Focusing on the 5E's of renal rehabilitation developed by the Life Options Rehabilitation Advisory Council, Renal Care Group of the Midwest, Inc has achieved positive clinical patient outcomes, such as meeting dialysis adequacy benchmark goals. Interventions and outcomes of two of the 5E's, education and exercise, are discussed. PMID- 10452713 TI - Designing a comprehensive hospital-wide patient education program. AB - Patient education is an integral part of any hospital admission. This article describes the development of a hospital-wide program which includes the assessment of patient education needs, the role of the unit patient educators, the use of free patient education television channels, the role of the patient/family education committee, the documentation of patient education, and the orientation of health professionals to the system. Mechanisms to deal with the non-English or limited-English-speaking patients are also described. PMID- 10452714 TI - Computed tomography (CT) today. PMID- 10452715 TI - Perfusion measurements of the brain: using dynamic CT for the quantitative assessment of cerebral ischemia in acute stroke. AB - OBJECTIVE: Perfusion CT has been successfully used as a functional imaging technique for the differential diagnosis of patients with hyperacute stroke. We investigated to what extent this technique can also be used for the quantitative assessment of cerebral ischemia. METHODS AND MATERIAL: We studied linearity, spatial resolution and noise behaviour of cerebral blood flow (CBF) determination with computer simulations and phantom measurements. Statistical ROI based analysis of CBF images of a subset of 38 patients from a controlled clinical stroke study with currently more than 75 patients was done to check the power of relative cerebral blood flow (rCBF) values to predict definite infarction and ischemic penumbra. Classification was performed using follow-up CT and MR data. RESULTS: Absolute CBF values were systematically underestimated, the degree depended on the cardiac output of the patients. Phantom measurements and simulations indicated very good linearity allowing reliable calculation of rCBF values. Infarct and penumbra areas in 19 patients receiving standard heparin therapy had mean rCBF values of 0.19 and 0.62, respectively. The corresponding values for 19 patients receiving local intraarterial fibrinolysis were 0.18 and 0.57. The difference between infarct and penumbra values was highly significant (P < 0.0001) in both groups. No penumbra area was found with an rCBF value of less than 0.20. While in the heparin group only 25% of all areas with an rCBF between 0.20 and 0.35 survived, in the fibrinolytic group 61% of these areas could be saved (P < 0.05). CONCLUSION: Perfusion CT is a fast and practical technique for routine clinical application. It provides substantial and important additional information for the selection of the optimal treatment strategy for patients with hyperacute stroke. Relative values of cerebral blood flow discriminate very well between areas of reversible and irreversible ischemia; an rCBF value of 0.20 appears to be a definite lower limit for brain tissue to survive an ischemic injury. PMID- 10452716 TI - Blood-brain barrier and blood volume imaging of cerebral glioma using functional CT: a pictorial review. AB - We present five cases of cerebral glioma that illustrate the benefit of functional CT imaging of blood-brain barrier permeability and cerebral blood volume. Functional CT uses Patlak analysis of a single location dynamic sequence to extract physiological information that is useful clinically in the assessment of cerebral gliomas. Functional CT offers distinct advantages over other functional modalities, including clearer delineation of tumour, tumour grading, measurement of tumour activity and monitoring response to therapy. PMID- 10452717 TI - Functional CT of the kidney. AB - The iodinated contrast agents used for computed tomography (CT) are filtered at the glomerulus and not reabsorbed by the tubules and have pharmacokinetics comparable to inulin. They can thus measure physiological indices such as contrast clearance per unit volume, which is closely related to glomerular filtration rate per unit renal volume of kidney, after due allowance for the difference between blood and plasma clearance. In this review, we show how dynamic CT can be used to measure both regional and global blood clearance of contrast material. A single slice of kidney is scanned sequentially after bolus intravenous (i.v.) injection of contrast material. Next, time-attenuation curves are constructed and contrast clearance per unit volume is calculated using a Patlak graphical analysis. CT determination of renal volume is made and global contrast clearance can be then also calculated. In normal kidneys, clearance/volume averaged 0.49+/-0.11 ml min(-1) ml(-1) (mean +/- S.D.), and these values agreed with literature data obtained using other techniques. A negative correlation between patient's age and clearance/volume was seen. A strong correlation was observed between creatinine whole blood clearance and the global contrast clearance (the product of renal volume determined by CT and contrast clearance/volume). Dynamic CT can provide quantitative renal physiological information on a regional basis non-invasively. PMID- 10452718 TI - Tumour angiogenesis and its relation to contrast enhancement on computed tomography: a review. AB - Angiogenesis describes the formation of new blood vessels within tumours. The process is essential for tumour growth and metastasis. The development of new vessels leads to physiological changes, specifically increased perfusion, blood volume and capillary permeability, that alter contrast enhancement during computed tomography (CT). Functional CT techniques that quantify these physiological changes can provide greater insight into how angiogenesis alters contrast enhancement in routine practice and also serve as diagnostic tools in their own right. The functional information obtained can aid with tissue characterisation, such as type or grade of tumour, improve the detection of hepatic metastases, produce clearer delineation of tumours with benefits for radiotherapy planning and biopsy, and provide prognostic information. By providing a marker for tumour angiogenesis, quantitative contrast enhanced CT can improve the diagnostic assessment of patients with cancer. PMID- 10452719 TI - Hepatic metastases: the value of quantitative assessment of contrast enhancement on computed tomography. AB - OBJECTIVE: Occult and overt hepatic metastases have been the target of research in an effort to improve detection and characterisation of cancer spread and, consequently, guidance of treatment. This paper aims to illustrate the value of two quantitative techniques for assessing contrast enhancement during CT in the detection of hepatic metastases. It outlines the applications to which they can be put, and the ease of incorporation into current protocols. METHODS AND MATERIAL: The first technique, perfusion CT, uses a single location dynamic CT sequence to obtain time attenuation data whilst a short, high concentration IV bolus of contrast passes through the abdominal vasculature. Quantitative hepatic arterial and portal values are calculated, along with a perfusion image map. The second technique uses densitometric analysis during a modified contrast enhanced dual-phase liver CT examination. Semi-quantitative values are calculated from the images obtained at the 25 and 40 s times. RESULTS: Both perfusion CT and densitometric analysis have been to shown to differentiate between normal and tumour-bearing liver as defined by structural CT. Hepatic metastases are associated with increased arterial perfusion and arterial phase enhancement. Increased arterial phase enhancement on densitometric analysis in the absence of overt lesions heralds the onset of visible metastases in the liver in the ensuing 18 months. Perfusion CT has also demonstrated a correlation between high arterial perfusion around a visible metastasis and increased survival. CONCLUSION: Both techniques can provide more information than is available from conventional enhanced CT scans alone. An algorithm for the clinical application of perfusion CT is proposed. The ease with which these quantitative techniques can be performed and the extra information they provide could lead to improved staging of cancer and more appropriate patient management. PMID- 10452720 TI - Functional renal perfusion imaging with colour mapping: is it a useful adjunct to spiral CT of in the assessment of abdominal aortic aneurysm (AAA)? AB - AIM: To ensure optimal timing with pre-operative spiral CT for abdominal aortic aneurysms (AAA), an initial 'timing' single level CT is commonly performed with a small bolus of contrast. This can be exploited to obtain adjunct functional information on renal perfusion. We have investigated the potential of this to measure renal perfusion, to produce colour renal perfusion maps and to predict surgical outcome in infrarenal aortic aneurysm assessment. METHODS: We studied 21 patients being assessed for repair of infrarenal AAA. Prior to the spiral CT, a single level through the renal hili and aorta was scanned after the intravenous injection of 25 ml of contrast given at 10 ml/s. Ten 1 s duration scans were performed from 8 to 30 s after injection. Optimal timing for CT angiography can then be determined. Time-density curves were then drawn for both kidneys and aorta using regions of interest (ROIs) or pixel-by-pixel analysis. Renal cortical perfusion was measured using both ROI analysis and pseudocolour perfusion images. Following previous work, perfusion was calculated as the peak upslope of the tissue time density curve divided by peak aortic enhancement. RESULTS: Cortical mean perfusion averaged 2.48 ml/min per ml (range 0.8-3.7 ml/min per ml n = 34) and the values obtained agreed with literature expectations. Follow up in the 10 patients proceeding to AAA repair suggest low mean perfusion values and predict a raised postoperative creatinine (P < 0.05) CONCLUSIONS: Additional functional data and imaging can be obtained from the initial timing scan of a CT study, without requiring a dedicated study. PMID- 10452721 TI - Imaging of tumour therapy responses by dynamic CT. AB - The objective of this study was to investigate whether functional CT with Patlak analysis could be used to demonstrate acute changes associated with radiotherapy. Patlak analysis yields fractional vascular volume and contrast clearance per unit volume (a measure of permeability). Four tumour types (prostate, bronchus, breast and cervix) were studied pre-radiotherapy and at 1-2 weeks and 6-12 weeks post therapy. Significant rises in fractional vascular volume and contrast clearance were shown at 1-2 weeks. These indices were still significantly elevated at 6-12 weeks post-therapy. In the prostates perfusion values were also elevated reflecting a hyperemic response to radiotherapy. Dynamic CT with Patlak analysis can be used to measure important pathophysiological indices which may prove useful in assessing response to therapy of tumours. PMID- 10452722 TI - Quiz case 8. Diagnosis: intraspinal hypertrophic callus formation in ankylosing spondylitis. PMID- 10452723 TI - Image-guided core breast biopsy of ductal carcinoma in situ presenting as a non calcified abnormality. AB - OBJECTIVE: Ductal carcinoma in situ (DCIS) typically presents as calcifications which are detected mammographically. Our aim was to evaluate the less common presentations of ductal carcinoma in situ diagnosed by image-guided core biopsy and correlate with histopathologic diagnoses. METHODS AND MATERIAL: Imaging and histopathologic findings were retrospectively reviewed in 11 patients with ductal carcinoma in situ diagnosed at core biopsy that presented as noncalcified radiographic abnormalities. RESULTS: Mammography showed non-calcified, circumscribed nodules, ill-defined nodules and architectural distortion. In two patients, no mammographic abnormality was detected. Sonography showed circumscribed, round or oval, solid masses; irregular, heterogeneous masses; and a tubular structure. Histopathologic diagnoses included multiple architectural subtypes and ranged from low to high nuclear grade. CONCLUSION: Although image guided core biopsy diagnosis of ductal carcinoma is typically made when sampling calcifications, DCIS can be diagnosed following biopsy of non-calcified masses or distortion. There is no correlation between histopathologic subtype and radiologic appearance. PMID- 10452724 TI - The value of T2(*)-weighted MR images for the diagnosis of acoustic neuromas. AB - The aim of this study was to evaluate high resolution T2(*)-weighted MRI--in our case a 3D CISS sequence--for the diagnosis of acoustic neuromas. Especially to be clarified was if high-resolution T2-weighted sequences can substitute T1-weighted contrast-enhanced sequences, and in which circumstances they can give important additional information when compared with contrast-enhanced T1-weighted sequences. METHODS AND PATIENTS: The MR examinations were performed using a 1.5 Tesla unit with a circularly polarized head coil. All 20 patients (11 females, nine males, aged from 12 to 80 years) with acoustic neuromas underwent preoperative MRI with T2*-weighted 3D CISS (slice thickness, 0.7 mm; acquisition time, 8 min) and pre- and postcontrast T1-weighted 3D MP-RAGE (slice thickness, 0.9 mm; acquisition time, 8 min) sequences. The detectability of acoustic neuromas was evaluated following the consensus of three radiologists using a 3D work station (parameters: tumor presence, extent, nerve attribution). RESULTS: All tumors were detected by both contrast-enhanced 3D MP-RAGE and 3D CISS and the diameters of the lesions were equally well measured. 3D CISS was the best sequence for the attribution of a lesion to a certain nerve. Labyrinthine involvement could be better detected using 3D CISS than contrast-enhanced 3D MP RAGE. CONCLUSION: High-resolution T2(*)-weighted MRI is a very sensitive method for tumor screening which can also detect even small meatal and labyrinthine neuromas. In the case of abnormal findings (other pathology or variations, e.g. vascular loops); however, contrast-enhanced T1-weighted MRI is necessary in order to confirm the presence of a tumor with typical enhancement. PMID- 10452725 TI - Gastric ulcer bleeding: diagnosis by computed tomography. AB - A case of CT demonstration of a bleeding gastric ulcer is presented, in a patient with confusing clinical manifestations. Abdominal CT was performed without oral contrast medium administration, and showed extravasation of intravenous contrast into a gastric lumen distended with material of mixed attenuation. It is postulated that if radiopaque oral contrast had been given, peptic ulcer bleeding would probably have been masked. CT demonstration of gastric ulcer bleeding, may be of value in cases of differential diagnostic dilemmas. PMID- 10452726 TI - Vitamin A deficiency in Bangladesh: a review and recommendations for improvement. AB - OBJECTIVE: This article provides a comprehensive review of the change in vitamin A status and the extent of vitamin A deficiency among different population groups in Bangladesh up to the present time. The result of experience with different strategies and interventions designed to improve vitamin A status are then reviewed, leading to a discussion of key options for action, as well as important areas for research and evaluation. DESIGN AND SETTING: All the available data have been examined in detail, including data from nationally representative samples and nationwide surveys, as well as small studies in different population groups. Reports on the effectiveness of different intervention programmes have been used. RESULTS: Over the past three decades a number of studies, which include national nutrition surveys, have been carried out to investigate the prevalence of vitamin A deficiency among different population groups in Bangladesh, and they have demonstrated a significant public health problem. Studies have shown that the prevalence of severe deficiency, based on the prevalence of night blindness in preschool children, decreased from 3.6% in 1982 83 to 1.78% in 1989 and 0.6% in 1996. However, there is still a high prevalence of subclinical vitamin A deficiency, based on the biochemical assessment of serum retinol levels in preschool children, estimated mainly from hospital-based groups. Night blindness and Bitot's spot are also found to exist among school-age children and adolescents. Recent reports indicate that night blindness among rural mothers is as high as 1.4%. Only a limited number of studies, with small sample sizes, are available where serum retinol has been reported for school-age children, adolescents and pregnant women. Nevertheless, these studies confirm the presence of low levels of serum retinol and hence, the existence of subclinical vitamin A deficiency. Furthermore, the dietary intake of vitamin A in each population group has been found to be less than the Recommended Daily Allowance (RDA), indicating a significant risk of deficient intakes of vitamin A. To address the problem of vitamin A deficiency, the government of Bangladesh started the Nutritional Blindness Programme in 1973. The main activities of the programme include vitamin A capsule (VAC) supplementation to children of 6 months to 6 years old, nutrition education to increase the production and consumption of vitamin A rich foods, and training of primary health-care workers on the clinical diagnosis and treatment of vitamin A deficiency, VAC distribution and nutrition education. Since 1988, as a long-term strategy, Helen Keller International has been implementing community home gardening promotion projects. To date, the possibility that foods may be fortified with vitamin A, has not been explored as a possible approach in Bangladesh. CONCLUSION: Although short- to long-term prevention and control programmes are to some extent in place, to improve the situation of vitamin A deficiency, Bangladesh needs a more appropriate mix of interventions for the entire population. More operational research and evaluation are needed if a fully effective programme to alleviate the problem of vitamin A deficiency is to be developed. Finally, to achieve the goal of virtual elimination of vitamin A deficiency will require an integrated approach which brings together appropriate actions at every level, within and across the many sectors of society. PMID- 10452727 TI - Use of medicines by older people in a large British national survey, and their relation to vitamin status indices. AB - OBJECTIVE: To describe the extent of use of medicines by older people living in Britain, and to explore relationships with vitamin status indices. DESIGN: The National Diet and Nutrition Survey of people aged 65 years and over collected data from a British sample during 1994-95. The present study has made novel use of it, to relate medicine use with biochemical indices of vitamin status. SETTING: Eighty postcode areas, selected randomly from mainland Britain. SUBJECTS: 1506 people, 65 years and over gave information about use of medication, and a 4-day weighed food record. Three-quarters gave blood for status indices. RESULTS: 78% of those living in the community and 93% of those in institutions were using medication. Certain vitamin status indices: plasma retinol, erythrocyte folate and riboflavin, paradoxically suggested better status in users than in non-users of antihypertensive, gastrointestinal, central nervous system, corticosteroid or diabetic drugs. There was evidence of a link with renal insufficiency, especially for plasma retinol, but neither this nor increased nutrient intake, acute phase effects or haemoconcentration could explain the paradoxical associations. CONCLUSIONS: Caution is needed in interpreting certain vitamin status indices, especially in older people who are extensively using medicines. New vitamin indices are needed, to avoid confounding interferences. PMID- 10452728 TI - Assessing the prevalence of nutrient inadequacy. AB - OBJECTIVE: To describe an approach for assessing the prevalence of nutrient inadequacy in a group, using daily intake data and the new Estimated Average Requirement (EAR). DESIGN: Observing the proportion of individuals in a group whose usual intake of a nutrient is below their requirement for the nutrient is not possible in general. We argue that this proportion can be well approximated in many cases by counting, instead, the number of individuals in the group whose intakes are below the EAR for the nutrient. SETTING: This is a methodological paper, and thus emphasis is not on analysing specific data sets. For illustration of one of the statistical methods presented herein, we have used the 1989-91 Continuing Survey on Food Intakes by Individuals. RESULTS: We show that the EAR and a reliable estimate of the usual intake distribution in the group of interest can be used to assess the proportion of individuals in the group whose usual intakes are not meeting their requirements. This approach, while simple, does not perform well in every case. For example, it cannot be used on energy, since intakes and requirements for energy are highly correlated. Similarly, iron in menstruating women presents some difficulties, due to the fact that the distribution of iron requirements in this group is known to be skewed. CONCLUSIONS: The apparently intractable problem of assessing the proportion of individuals in a group whose usual intakes of a nutrient are not meeting their requirements can be solved by comparing usual intakes to the EAR for the nutrient, as long as some conditions are met. These are: (1) intakes and requirements for the nutrient must be independent, (2) the distribution of requirements must be approximately symmetric around its mean, the EAR, and (3) the variance of the distribution of requirements should be smaller than the variance of the usual intake distribution. PMID- 10452729 TI - What is the research base for the use of dietary supplements? AB - The market for dietary supplements in the USA was estimated as about 11.8 billion dollars in 1997 with a growth rate of 10-14% projected in the next 3 years. Data from the Food and Drug Administration collected in 1995 indicate that over 55% of adults surveyed used some type of dietary supplement. The marketing of dietary supplements in the USA has been essentially deregulated by the passage of the Dietary Supplement Health and Education Act of 1994 (DSHEA). This legislation defined dietary supplements, made manufacturers responsible for the safety of supplements and allowed certain statements of nutrition support to be made on supplement labels. The US Congress in passing the DSHEA indicated that supplements should be available on the market so that consumers could make decisions about their use for themselves and their families. Unfortunately, information about the research base for supplement claims is not readily accessible to health professionals and consumers. There is a need for authoritative reviews of the data underlying supplement claims to assist public health professionals in their role of providing advice to the public about dietary supplements. PMID- 10452730 TI - Development and testing of a quantitative food frequency questionnaire for use in Gujarat, India. AB - OBJECTIVE: To develop and test a quantitative, interviewer-administered food frequency questionnaire (FFQ) to ascertain nutrient intakes of individuals in northern India. DESIGN: A 92-item FFQ was developed based on food use and market surveys of the study area. A validation study was conducted consisting of 24-h diet recalls (24HR) administered on 6 randomly selected days over 1 year. Two FFQs were administered, one each at the beginning and end of the 1-year period. FFQ and 24HR-derived nutrient scores were compared using correlation and regression analyses and by computing differences between nutrient intakes estimated by the two methods. SETTING: Rural villages in Bhavnagar District, Gujarat, North India. SUBJECTS: 60 individuals who agreed to provide all necessary data. RESULTS: Pearson (parametric) correlation coefficients averaged 0.69 in comparing nutrient scores derived from the 24HR with those from the first FFQ and 0.72 in comparing the second FFQ (P<0.0001). Spearman correlation coefficients were virtually identical to the Pearson correlations, averaging 0.68 and 0.72, respectively. In regression analyses, most coefficients were close to 1.0 (perfect linear association). Nutrient scores were significantly and consistently higher on both FFQs relative to the 24HR. CONCLUSIONS: This FFQ produces results broadly comparable, and superior in some respects, to those commonly used in the West. Higher than average measures of association indicate its suitability for comparing exposures within this study population in reference to health-related endpoints. PMID- 10452731 TI - Plasma vitamin C: what does it measure? AB - OBJECTIVE: To examine the association between self-reported consumption of foods and plasma vitamin C levels. DESIGN: A cross-sectional analysis of dietary data and plasma vitamin C levels. Subjects placed the following foods into frequency categories: fresh fruit, leafy greens, other vegetables, fatty fish, other fish, chicken, meat, meat products, eggs, cheese and brown bread. The six frequency categories ranged from 'never' to 'at least daily'. Plasma vitamin C was measured by fluorometric assay. SETTING: A population-based cohort study in Norfolk, UK. SUBJECTS: 598 men and 566 women aged 45-74 years not taking vitamin supplements. RESULTS: Plasma vitamin C was positively correlated with intake of fresh fruit (r=0.29 in men and r=0.25 in women, P<0.001), leafy greens (r=0.20 in men P<0.001, r=0.13 in women P<0.01), other vegetables (r=0.20 in men P<0.001, r=0.14 in women P<0.01) and brown bread (r=0.28 in men, r=0.17 in women, P<0.001) and negatively associated with intake of meat products (r=-0.13 in men P=0.02, r= 0.10 in women P<0.01). The difference in plasma vitamin C between never and daily eaters of brown bread was 13.6 micronol l(-1) in men and 9.9 micromol l(-1) in women, P<0.001. CONCLUSIONS: These data suggest that plasma vitamin C is not only a marker of foods rich in vitamin C but of certain patterns of food consumption. Such patterns are likely to be population specific and might explain inconsistencies in biomarker-disease associations. PMID- 10452732 TI - Dietary intake of Indians living in the metropolitan area of Durban. AB - OBJECTIVE: To report on energy and nutrient intakes, as a risk factor for coronary heart disease in Indian South Africans. DESIGN: Cross-sectional dietary study as part of a coronary heart disease survey. SETTING: Metropolitan area of Durban, South Africa. SUBJECTS: Free-living Indian men (n = 406) and women (n = 370) 15-69 years of age. METHODOLOGY: Dietary data were collected by three interviewers using a 24-h dietary recall and expressed as median intakes of macronutrients. RESULTS: Results reported a low energy intake and the percentage of energy derived from total fat varied between 32.3 and 34.9% in men and between 33.1 and 36.1% in women. The energy intake to basal metabolic rate (EI:BMR) ratios were low for all age groups suggesting potential under-reporting. Intake of polyunsaturated fatty acids was high, more than 10% of energy, with a median polyunsaturated to saturated fatty acid ratio (P:S ratio) that varied between 1.38 and 1.96 for the various age and sex groups. Dietary cholesterol varied between 66 and 117 mg per 4.2 MJ in men and between 76 and 109 mg per 4.2 MJ in women. Dietary fibre intakes were low and varied between 8.0 and 11.0 g per 4.2 MJ in men and between 7.6 and 9.6 g per 4.2 MJ in women. CONCLUSIONS: The dietary P: S ratios were high and the effect of such a high P:S ratio on the oxidation of low density lipoprotein in this population, with a high prevalence of coronary heart disease (CHD), should be investigated as a possible risk factor for CHD. PMID- 10452733 TI - Assessment of a narrative approach to the diet history. AB - OBJECTIVE: To assess the quality of a narrative form diet history (DH). DESIGN: Reproducibility assessed with data obtained at 6-week intervals. Criterion validity assessed using energy intake to estimated energy expenditure (EI:BMR) cutoff limits. Relative validity assessed by comparing results for energy and macronutrients at baseline and month 2 of an intervention trial with those obtained from 3-day food records (FR). SETTING: Community-based dietary intervention trials for the study of metabolic syndrome in the Illawarra region of New South Wales, Australia. SUBJECTS: Reproducibility: 43 healthy female volunteers. Mean age 58.72 years (range 50-67), mean body mass index (BMI) 25.79 (range 21-36). VALIDITY: 45 healthy volunteers--18 males (mean age 46.9 years, mean BMI 27.8), and 27 females (mean age 45.7 years, mean BMI 26.2), attending a study on the effect of diet on metabolic variables. RESULTS: Reproducibility: wide SD values indicated a high degree of intraindividual variation, but correlation coefficients were comparable to those of similar studies. VALIDITY: underreporting was inconsistent with individuals, but was greater with the DH than the FR at each stage of the dietary trial (significant at month 1, P < 0.01). Underreporters indicated higher intakes of protein during the trial (P < 0.05). Weaker associations were found between the DH and FR data for energy values, but there was strong agreement for per cent fat consumed as saturated and monounsaturated fat at each stage of the trial. CONCLUSIONS: The narrative form DH performs as well as standardized interviews, but more work needs to be done at the micro level, focusing on aspects which deal with foods likely to be underreported in the particular study context. This can be achieved through continued research using combined methodologies. PMID- 10452734 TI - Diet and lifestyle characteristics associated with dietary supplement use in women. AB - OBJECTIVES: To describe the characteristics of dietary supplement users in a large cohort of women and test the hypothesis that supplement users would be more likely to have a healthier lifestyle than non-users. DESIGN: Comparison of nutrient intakes from food frequency questionnaire (FFQ) data for 8409 supplement users and 5413 non-users. Use of logistic regression modelling to determine predictors of supplement use in this cohort. SUBJECTS: 13,822 subjects from the UK Women's Cohort Study (UKWCS) for whom data on supplement use was available. RESULTS: Significant differences in nutrient intakes from FFQ were seen between the two groups, with supplement users having higher intakes of all nutrients, except for fat and vitamin B12. Use of dietary supplements was associated with being vegetarian, vegan or fish-eating, consuming more fruit and vegetables, being more physically active and having a lower alcohol intake. Supplement use was less likely in those with a body mass index above 25 and those who reported smoking regularly. CONCLUSIONS: The findings are consistent with the hypothesis that supplement use is associated with a healthier lifestyle profile and an adequate nutritional intake, suggesting that supplement users do not need to take supplements to meet a nutrient deficiency. PMID- 10452735 TI - Hopes towards 'healthy eating' [correction of 'healthy heating']: are we being realistic? PMID- 10452736 TI - Protozoan paradigms for cell biology. PMID- 10452738 TI - Gastroesophageal reflux and asthma: insight into the association. AB - Gastroesophageal reflux (GER) is a potential trigger of asthma. GER symptoms are more prevalent in asthma patients compared with control populations, with a prevalence of approximately 75%. GER symptoms are associated with respiratory symptoms and inhaler use. GER may also occur without esophageal symptoms. Abnormal esophageal acid contact times are also more prevalent in patients with asthma compared with control populations, with a prevalence of 80%. Pathophysiologic mechanisms of esophageal acid-induced bronchoconstriction include a vagally mediated reflex, heightened bronchial reactivity, and microaspiration. Esophageal acid may increase minute ventilation without evidence of bronchoconstriction. Esophageal acid is associated with the release of substance P in the bronchial mucosa, resulting in airway edema. Medical antireflux therapy with proton pump inhibitors results in asthma symptom improvement in approximately 70% of patients, similar to surgical results. Predictors of asthma response include the presence of regurgitation, proximal acid reflux, esophagitis healing with therapy, reflux-associated respiratory symptoms, or nocturnal asthma. Management of GER in adult patients with asthma should include a 3-month trial of high-dose proton pump inhibitor while monitoring asthma outcome. GER should be considered as a potential asthma trigger in all patients. PMID- 10452739 TI - Oligonucleotide therapy of allergic asthma. AB - The recent increase in the prevalence of and mortality from asthma has inspired several new molecular techniques to improve treatment. Because asthma is a disease of gene polymorphism, gene therapy is unlikely to be effective. Alternative methods use oligonucleotides (ODNs) in the form of (1) DNA vaccination expressing CpG motifs that mimic bacterial DNA or (2) antisense ODNs inhaled and locally deposited into pulmonary airways to specifically modulate receptors for inflammatory mediators. DNA vaccination, a form of "molecular immune surveillance," attenuates a TH2 predominance. Antisense directed against the adenosine A(1 ) receptor abrogates A(1 ) sensitivity, improves allergen induced immediate airway obstruction, and inhibits the expected increase in histamine responsiveness in allergic rabbits. Adenosine receptor inhibition lasts for an average of 7 days and the majority of the antisense remains in the lung. ODN therapy for asthma seem unlimited, but confirmation awaits the extension from animal models to human studies. PMID- 10452740 TI - Sublingual immunotherapy. PMID- 10452741 TI - Anaphylaxis in the general population: A frequent and occasionally fatal disorder that is underrecognized. PMID- 10452745 TI - Syk deficiency in nonreleaser basophils. AB - BACKGROUND: Peripheral blood basophils from 10% to 20% of donors fail to degranulate in response to cross-linking the high-affinity IgE receptor FcepsilonRI. The molecular mechanisms underlying the nonreleaser phenotype have not been established. OBJECTIVES: Our aim was to compare the expression of FcepsilonRI-associated protein tyrosine kinases between nonreleaser and releaser basophils. METHODS: With use of Western blotting we investigated Syk and Lyn protein levels in highly purified basophils from 3 anti-IgE nonreleasers and 2 releasers. RESULTS: We identified 3 healthy nonatopic donors whose nonreleaser basophils express FcepsilonRI normally but fail to express protein for the tyrosine kinase Syk, which is implicated in the initiation of FcepsilonRI mediated secretion. Protein levels for the tyrosine kinase Lyn are somewhat reduced but not absent in nonreleaser basophils. Levels of Lyn and Syk protein are similar in B cells, eosinophils, and neutrophils from releaser and nonreleaser donors. During these studies one nonreleaser "converted" into a releaser with concomitant basophil Syk expression. CONCLUSION: The absence of detectable Syk could explain the nonreleaser phenotype of basophils from some donors. PMID- 10452746 TI - Warming of feet elevates nasal mucosal surface temperature and reduces the early response to nasal challenge with allergen. AB - BACKGROUND: We have previously shown that hot, humid air partially reduces the early allergic response. Mechanisms for this effect have been suggested, but none has gained universal acceptance. The most likely explanations are a modification of mucosal temperature or a reduction in nasal secretion osmolality. OBJECTIVE: We sought to investigate whether increasing the nasal mucosal surface temperature by immersing feet in warm water (WW) could decrease the immediate nasal response to challenge with allergen. METHODS: We performed a randomized, 2-way crossover study on 14 subjects with seasonal allergic rhinitis outside of their allergy season. They immersed their feet in either WW (42 degrees C) or room-temperature water (RW; 30 degrees C) for 5 minutes before and during nasal challenge with diluent for the allergen extract, followed by 2 increasing doses of allergen. RESULTS: There was a statistically significant increase in nasal mucosal temperature from baseline after warming of feet (WW, 1.9 +/- 0.1 degrees C, vs RW, 0.2 +/- 0.1 degrees C; P =. 001), but there were no significant differences in body temperature (WW, 0.1 +/- 0.1 degrees C, vs RW, 0.4 +/- 0.1 degrees C; P =.1). Net changes from diluent challenge for all parameters were compared between immersion of feet in WW and RW. Immersion of feet in WW significantly inhibited allergen-induced sneezes (WW, 5.7 +/- 1.1, vs RW, 11.6 +/- 3.2; P <.01), human serum albumin levels (WW, 941.7 +/- 172.2 microg/mL vs RW, 1524.8 +/- 220.6 microg/mL; P <.01), and secretion weights (WW, 30.5 +/- 7.2 mg, vs RW, 41.8 +/- 6.8 mg; P <. 01). CONCLUSION: Our data show that warming of feet decreases the early response to nasal challenge with antigen. This inhibitory effect is probably related to the increase in the nasal mucosal temperature. PMID- 10452747 TI - Proteasome inhibition: A novel mechanism to combat asthma. AB - BACKGROUND: Nuclear factor-kappaB (NF-kappaB) is a critical transcription factor required for the regulation of many genes involved in inflammatory responses to noxious stimuli. On activation, NF-kappaB induces the transcription of numerous proinflammatory cytokines, enzymes, and cellular adhesion molecules. Blockade of the proteasome with selective inhibitors attenuates the effects of NF-kappaB, leading to suppression of the inflammatory response. OBJECTIVE: We sought to determine whether proteasome inhibitors would be active in a model of asthma. METHODS: The mouse delayed-type hypersensitivity model was used to screen a panel of compounds for in vivo activity. The proteasome inhibitor, PS-519, was shown to be the most active in this model and was selected for further development. Allergen-induced pulmonary eosinophilia in Brown Norway rats was used subsequently to determine anti-inflammatory activity in an animal model. RESULTS: Direct administration of PS-519 into the lungs significantly reduced leukocyte numbers, particularly the selective increase in eosinophils. Because steroids are the mainstay anti-inflammatory therapy in asthma, and data is available to suggest their possible interaction to suppress the activation of NF-kappaB, rats were also treated by inhalation with combinations of a steroid and the proteasome inhibitor. In both the delayed-type hypersensitivity and the animal eosinophil model, low doses of proteasome inhibitors were shown to be effective when given with low doses of steroids. CONCLUSION: Taken together, the present data suggest that proteasome inhibition may represent a novel strategy for the treatment of inflammatory lung diseases such as asthma. PMID- 10452748 TI - Perennial rhinitis: An independent risk factor for asthma in nonatopic subjects: results from the European Community Respiratory Health Survey. AB - BACKGROUND: Although clinical and experimental studies suggest that upper respiratory tract dysfunction may affect the lower airways, rhinitis is usually not studied as a potential risk factor for asthma. This is because both diseases share key elements of pathogenesis and are usually considered as different manifestations of the same underlying "atopic" state. OBJECTIVE: We sought to assess whether asthma is associated with rhinitis in the absence of immunologic disorders in a population study. METHODS: Data from 34 centers participating in the European Community Respiratory Health Survey were analyzed. Random samples of 20- to 44-year-old subjects were invited to complete a detailed questionnaire and undergo total and specific IgE measurements, skin prick tests to 9 allergens, and bronchoprovocation challenges with methacholine. RESULTS: Subjects with perennial rhinitis (n = 1412) were more likely than control subjects (n = 5198) to have current asthma. After adjustment for sex, age, smoking habit, family history of asthma, geographic area, and season at the time of examination, asthma was strongly associated with rhinitis among atopic subjects (odds ratio [OR] = 8.1; 95% confidence interval [CI] = 5.4-12.1) but also among nonatopic subjects (OR = 11.6; 95% CI = 6.2-21.9). Moreover, the association remained very strong when the analysis was restricted to nonatopic subjects with IgE levels of 80 kIU/L or less (OR = 13.3; 95% CI = 6. 7-26.5). In nonasthmatic subjects bronchial hyperresponsiveness was also more frequent in subjects with rhinitis than in those without rhinitis (OR = 1.7; 95%CI = 1.2-2.6 in nonatopic subjects with IgE levels of 0.999) over a wide range of CA concentrations (5-400 pmol). The limit of detection of CA in plasma using this method is <0.1 microM and only 50 microl of plasma is required for the assay. By coupling this method with a recently described HPLC-fluorescent method to determine acrolein, a cytochrome P450 metabolite of IFA formed during the activation of the drug by 4-hydroxylation, the two major, alternative P450 catalyzed pathways of IFA metabolism can be monitored from the same plasma samples or liver microsomal incubations and the partitioning of drug between these two pathways thereby quantitated. This assay may prove to be useful for studies of IFA metabolism aimed at identifying factors that contribute to individual differences in CA formation and in developing approaches to minimize CA formation while maximizing IFA cytotoxicity. PMID- 10452808 TI - A myeloperoxidase-specific assay based upon bromide-dependent chemiluminescence of luminol. AB - Measurement of myeloperoxidase (MPO; EC 1.11.1.7) activity is often used as a marker of neutrophil infiltration into tissues. However, most enzymatic assays for MPO are susceptible to interference from other peroxidases (including eosinophil peroxidase, EPX) and hemoproteins (such as hemoglobin and myoglobin) present in the tissues. In this report, we describe a bromide-dependent chemiluminescence (Br-CL) assay that uses luminol as a chemiluminescence probe. The assay can distinguish between MPO and nonspecific peroxidase reactions. The MPO-specific reaction is believed to proceed in two steps: (i) the enzymatic generation of hypobromous acid (HOBr) from KBr and H(2)O(2) at pH 5 and (ii) the spontaneous reaction of HOBr and H(2)O(2) with luminol to give a Br-CL signal. The assay is sufficiently sensitive to allow detection of MPO in <100 human neutrophils. Other peroxidases and hemoproteins do not interfere with the Br-CL signal. Although EPX can also oxidize bromide to generate HOBr, activities of MPO and EPX can be distinguished at different pHs. As a demonstration of the utility of the Br-CL assay, MPO activity was measured in murine tumors known to be infiltrated with neutrophils. A statistically significant correlation was seen between MPO activity and histological neutrophil counts in the tumors (r = 0.69, P < 0.01, n = 14). The assay should have wide application for measuring the neutrophil content of tissues. PMID- 10452809 TI - Analysis of the binding of the Fab fragment of monoclonal antibody NC10 to influenza virus N9 neuraminidase from tern and whale using the BIAcore biosensor: effect of immobilization level and flow rate on kinetic analysis. AB - The binding of the Fab fragment of monoclonal antibody NC10 to influenza virus N9 neuraminidase, isolated from tern and whale, was measured using an optical biosensor. Both neuraminidases, homotetramers of 190 kDa, were immobilized to avoid multivalent binding, and the binding of the monovalent NC10 Fab to immobilized neuraminidase was analyzed using the 1:1 Langmuir binding model. A contribution of mass transport to the kinetic constants was demonstrated at higher surface densities and low flow rates, and was minimized at low ligand densities and relatively high flow rates (up to 100 microl/min). Application of a global fitting algorithm to a 1:1 binding model incorporating a correction term for mass transport indicated that mass transport was minimized under appropriate experimental conditions; analysis of binding data with a mass transport component, using this model, yielded kinetic constants similar to those obtained with the 1:1 Langmuir binding model applied to binding data where mass transport had been minimized experimentally. The binding constant for binding of NC10 Fab to N9 neuraminidase from tern influenza virus (K(A) = 6.3 +/- 1.3 x 10(7) M(-1)) was about 15-fold higher than that for the NC10 Fab binding to N9 neuraminidase from whale influenza virus (K(A) = 4.3 +/- 0.7 x 10(6) M(-1)). This difference in binding affinity was mainly attributable to a 12-fold faster dissociation rate constant of the whale neuraminidase-NC10 Fab complex and may be due to either (i) the long-range structural effects caused by mutation of two residues distant from the binding epitope or (ii) differences in carbohydrate residues, attached to Asn(200), which form part of the binding epitope on both neuraminidases to which NC10 Fab binds. PMID- 10452810 TI - "Essays in the history of neurolinguistics". PMID- 10452811 TI - The 1874 aphasia debate in the Berliner Gesselschaft fur Anthropologie. PMID- 10452813 TI - Charcot's brains. PMID- 10452812 TI - Does Gustave Dax deserve to be forgotten? The temporal lobe theory and other contributions of an overlooked figure in the history of language and cerebral dominance. AB - Gustave Dax played an important role in the early history of cerebral dominance for language. He not only sent the 1836 memoir of Marc Dax, his deceased father, to Paris in 1863, but saw to it that this important document was published before Paul Broca's own article on cerebral dominance appeared later in 1865. In addition, he supported his father's contention that the left hemisphere is special for speech with 140 additional clinical cases. Gustave Dax's own unique contribution, however, has been almost completely overlooked. Although his theory lacked specificity, he preceded Meynert, Schmidt, and Wernicke in suggesting that the left temporal lobe may be especially important for speech. PMID- 10452814 TI - Falret, Seglas, Morselli, and Masselon, and the "language of the insane": a conceptual history. PMID- 10452815 TI - Freud's continuity thesis. PMID- 10452817 TI - Presence and localization of histidine decarboxylase enzyme (HDC) and histamine in Tetrahymena pyriformis. AB - Histidine decarboxylase (HDC) enzyme and its function under hormonal influences were studied in a low level of phylogeny. HDC protein is present in the unicellular ciliate Tetrahymena and its expression was not altered by insulin or histamine treatment. Starvation for 24 h enormously decreased the quantity of histamine in the cells. However, insulin influenced the activity of the HDC enzyme, demonstrated by the seven-fold quantity of histamine in the starved cells after insulin treatment. Insulin also increased the uptake of histamine from the tryptone-yeast extract medium. HDC was found in different parts of the cytoplasm, mainly in the periphery (epiplasm) of the cells. The experiments demonstrated the uptake and synthesis of histamine by Tetrahymena as well as the possibility of hormonal regulation of HDC activity. PMID- 10452816 TI - Apoptosis induced by filarial parasitic sheath protein in HEp 2 cell lines blocked by ectopic expression of bcl 2. AB - Chronic filarial patients exhibit an occult manifestation, Tropical Pulmonary Eosinophilia, (TPE), caused by an exaggerated immune response to shed and circulating filarial antigens, leading to extensive lung damage. We have attempted to examine the disease in vitro using the human epithelial cell line, HEp2. Filarial sheath proteins induce apoptosis in HEp2 cells characterized by chromatin condensation, internucleosomal DNA cleavage, positive staining for TUNEL assay and shows a sub-G1 peak on FACS analysis. In order to understand subcellular events and to analyse the protective role of bcl2, we engineered HEp2 to overexpress Bcl2 protein. HEp2 bcl2 cells do not undergo apoptosis on exposure to filarial sheath protein, indicating that filarial protein-induced apoptosis in epithelial cells proceeds via a pathway, inhibitable by overexpression of bcl 2. PMID- 10452818 TI - Effect of dexamethasone on the assembly of the matrix of fibronectin and on its receptors organization in Ehlers-Danlos syndrome skin fibroblasts. AB - The effect of dexamethasone (DEX) on the expression of fibronectin (FN), proalpha(1)(I) collagen (Col1), integrin alpha(2), alpha(5)and beta(1)subunits mRNAs, were studied by quantitative in situ hybridization (ISH) with radiolabelled probes in relationship with the organization of the extracellular matrix (ECM) of FN in human skin fibroblasts. In particular, two fibroblast strains were analysed, one derived from a control donor, typically organizing a rich ECM of FN, and the other from a patient affected by Ehlers-Danlos syndrome (EDS), which did not assemble the FN-ECM. Treatment of both fibroblast strains with 10(-7) m DEX slightly enhanced the level of FN mRNA (by about 1.5-fold), did not influence the level of alpha(5)subunit mRNA and reduced Col1, alpha(2)and beta(1)integrin subunits mRNAs by 2-3-fold. These results show that, in these cells, DEX coordinately downregulates the expression of Col1 and its specific integrin alpha(2)beta(1). Moreover, DEX regulates in a different manner the alpha(5)and beta(1)subunits forming the main FN receptor (FNR) in skin fibroblasts. Immunofluorescence microscopy evidencing the FN-ECM and integrins containing alpha(5)and beta(1)subunits showed that in control cells DEX induced a slight enhancement of the FN-ECM and of the alpha(5)beta(1)receptors patches. Therefore, in these cells the decrease of beta(1)FN receptor subunit mRNA, as well as the decrease of Col1 and its receptor mRNAs, did not influence the FN-ECM assembly. In EDS fibroblasts, DEX decreased the cytoplasmic accumulation of FN and induced the assembly of a rich FN-ECM through the formation of large FNR integrin patches, codistributing with the FN-ECM. We suggest that in EDS skin fibroblasts DEX corrects the defective FN-ECM favouring the sorting and the organization of FN and its alpha(5)beta(1)integrin receptor. PMID- 10452819 TI - The role of Ca ions in restoration of the structure of interphase and mitotic chromosomes in PK living cells after hypotonic stress. AB - The dynamics of mitotic chromosome and interphase chromatin recondensation in living PK cells during their adaptation to hypotonic medium was studied. The recondensation process was found to be slowed down by the modification of plasma membrane with low concentrations of glutaraldehyde, while osmotic reactions of glutaraldehyde-treated cells remain unchanged. The effect of glutaraldehyde can be rapidly reversed by the addition of Ca(2+)-ionophore A23187. Intracellular Ca(2+)measurements show that the adaptation to hypotonic shock is accompanied by restoration of free Ca concentration, whereas the delay of chromatin condensation in glutaraldehyde-treated cells is paralleled by the decrease of Ca level. The mechanisms implying the role of low concentration of Ca(2+)in chromatin compactization in vivo are discussed. PMID- 10452820 TI - Two-dimensional protein maps of xenopus eggs and embryos at different developmental stages. AB - Protein expression during the early development of Xenopus has been followed by 2D-polyacrylamide gel electrophoresis (PAGE). The analysis of two-dimensional maps of eggs and embryos at different stages of development has allowed the separation of more than 2000 spots. Identification of numerous polypeptides was obtained in four different ways: (1) immuno-blotting; (2) amino terminal sequence after blotting on to PVDF membranes; (3) comigration; and (4) assignment in comparison with proteins separated by 2D techniques on reference maps such as human liver, red blood cells, plasma and cerebrospinal fluid reported in the Swiss 2D-PAGE Data Base. The maps presented in this report are a step toward the study of the protein expression in Xenopus eggs and embryos and may be a powerful working tool since Xenopus embryos are popular models for the study of development. PMID- 10452821 TI - Expression of major gap junction connexin types in the working myocardium of eight chordates. AB - The alpha1 connexin (connexin43) is regarded as the major gap junction protein of the myocardium because it predominates there in mammals. Here, we show that it is not the major connexin of the working myocardium in non-mammalian vertebrates, which instead express beta1-like connexins homologous to mammalian connexin32. A phylogenetic series of hearts was immunostained with seven antibodies raised against peptide sequences specific for three distinct members of the gap junction connexin family: alpha1, beta1 and alpha5 (mammalian connexin40/avian connexin42). Working myocardium from two ascidian chordates (Ciona and Mogula), a teleost (Carassius), a frog (Xenopus) and two reptiles (Anolis and Alligator) was found to express a beta1-like connexin, rather than an alpha1-like connexin. An alpha1-like connexin was nevertheless often detected in other cardiac tissues. In the chicken (by ancestry a reptile), the developing myocardium expressed a beta1 like connexin strongly on embryonic day 6 but less strongly at hatching, and minimally in the adult. Myocardial expression of alpha5 connexin increased during development, but remained strongest in the coronary vascular endothelial and cardiac conduction tissues. The arteriolar smooth muscle of the chicken expressed alpha1 connexin throughout development, but its myocardium did not. In contrast, the working myocardium of a marsupial mammal (the opossum Trichosurus) strongly expressed an alpha1 connexin just like placental mammals. These results imply that a shift from beta1 to alpha1 connexin expression in the heart occurred prior to the evolution of the opossums. The beta and alpha connexin subfamilies have different permeabilities and gating properties, and we discuss factors that might have made this shift beneficial. PMID- 10452822 TI - Alterations in glycosyltransferases during myeloid and monocytoid differentiation of HL-60 cells. AB - HL-60, a human promyelocytic leukemia cell line, can be differentiated to myeloid lineage by all- trans retinoic acid (ATRA), dimethylsulfoxide (DMSO) and n butyric acid (n -BA), or to monocytoid(monocytic/macrophagic) lineage by phorbol 12-myristate-13-acetate (PMA) and ganglioside GM(3). The activity alterations of N -acetylglucosaminyltransferase III and V (GnT-III, GnT-V) as well as alpha-1,6 fucosyl-tranferase (alpha1,6 Fuc T) were studied during the differentiation of HL 60 cells by the above-mentioned five inducers using the fluorescence (PA)-labeled glycan-HPLC method for GnT assays and biotin-labeled glycan-LCA affinity chromatography combined with the HRP-avidin colorimetric method for alpha1,6 Fuc T assay. It was observed that after 3 days, all three enzymes decreased in HL-60 cells induced by 1 micromol/l ATRA and 0.6 mmol/l n-BA, while GnT-III and alpha1,6 Fuc T increased, but GnT-V still decreased after induction by 1% DMSO. GnT-V and alpha1,6 Fuc T declined, while GnT-III was elevated after induction by 0.1 micromol/l PMA for 3 days. In contrast, GnT-III increased after the treatment with 50 micromol/l GM(3)for 3 or 6 days, but GnT-V was not appreciably changed and alpha1,6 FucT was elevated after 6 days of GM(3)treatment. It may be concluded that the decrease of GnT-V is the common change in myeloid differentiation and the increase of GnT-III is the general alteration in monocytoid differentiation. The changes in the activities of glycosyltransferases were consistent with the structural changes in surface N -glycans previously found in our laboratory, i.e. that the antennary number of N -glycans decreased during myeloid differentiation by ATRA, and the amount of bisecting GlcNAc in N glycans increased during monocytoid differentiation by PMA. PMID- 10452823 TI - Expression of cation exchanger NHE and anion exchanger AE isoforms in primary human bone-derived osteoblasts. AB - The authors used isoform-specific antibodies against cation (NHE) and anion (AE) exchange isoforms in order to establish their specific expression and localization in dispersed human bone-derived cells. Immunocytochemical preparations of permeabilized osteoblasts probed with polyclonal antibodies were optically analysed by conventional immunofluorescence and con-focal laser scanning microscopy. These techniques demonstrated the abundant presence of epitopes of the cation exchangers NHE1 and NHE3 and the anion exchanger AE2 in these cells. The NHE1 and NHE3 isoform proteins were predominantly located in subplasmalemmal and nucleoplasmic vesicles. The AE2 isoform was densely localized to a subcellular location characteristic of the Golgi complex. The molecular identity of the AE and NHE isoforms was investigated by RT-PCR that confirmed the presence of NHE1 and NHE3 transcripts in addition to NHE4. RT-PCR and diagnostic restriction analysis of amplified AE cDNA established preferential AE2 expression. Since AE2 has been shown to act as a sulfate transporter at low pH, it is possible that it performs this function in the osteoblast Golgi complex where sulfation reactions occur post-translationally on numerous extracellular matrix macromolecules prior to secretion and mineralization. The Na(+)/H(+)exchanger proteins are regulated by mitogenic and non-mitogenic stimuli in the osseus environment and are involved in the large fluxes of ions and protons that necessarily occur during bone formation and resorption and thus play an important role in intracellular ion homeostasis in osteoblasts. PMID- 10452824 TI - Intracisternal A-particle (IAP)-mediated leukemogenesis: levels and stability of IAP mRNA in FDC-P1 cells exposed to the conditions of an irradiated environment. AB - Following injection into sublethally irradiated DBA/2 or BALB/c mice, factor dependent FDC-P1 cells undergo leukemic transformation due to oncogene activation by insertion of intracisternal A-particle (IAP) genetic elements. Similar events are observed in vitro during coculture of FDC-P1 cells with irradiated bone marrow stroma cells. To elucidate the mechanism of IAP transposition, we studied the level of IAP expression under the growth conditions preceding cell transformation. In vitro experiments showed that the type of growth factor, FDC P1 cell density, costimulation with steroid hormones or abrupt growth factor withdrawal had no effect on IAP mRNA levels (major transcripts of 7.4, 4.0 and 1.9 kb). By contrast, stimulation with suboptimal concentrations of GM-CSF or IL 3 induced a mean 2. 5-fold increase in the intensity of the 7.4 kb band, and induction of macrophage differentiation with retinoic acid resulted in an increased stability of the 4.0 kb band. Although suboptimal growth stimulation and incipient macrophage differentiation have previously been shown to occur in the process of FDC-P1 cell transformation, an increase in IAP expression could not convincingly be demonstrated in FDC-P1 cell populations isolated from irradiated BALB/c mice or stroma cell cocultures. Further experiments are required to define the role of suboptimal growth stimulation and/or macrophage differentiation in this transformation model. PMID- 10452825 TI - Changes in endogenous polyamine levels are associated with differentiation in Dictyostelium discoideum. AB - This is the first report correlating levels of polyamines and its fractions with differentiation in Dictyostelium discoideum. Temporal changes in endogenous levels of free, conjugated and bound putrescine, spermidine and spermine were analysed at critical stages of morphogenesis in this organism. No spermine was found at any given stage and putrescine was the most abundant polyamine. There was a sharp increase in the levels of both free (and total) and conjugated forms of putrescine and spermidine at the slug stage as compared to the growth phase. The levels of putrescine and spermidine were found to be higher in isolated prespore cells as compared to the prestalk cells. Remarkably, the levels of polyamine decreased at the early culminant stage. Data suggest that a moderate level of polyamines is needed for growth but it is important to have high levels of polyamines at the time of differentiation. PMID- 10452826 TI - Malignant transformation of Syrian hamster embryo (SHE) cells in primary culture by malachite green: transformation is associated with abrogation of G2/M checkpoint control. AB - Malachite green (MG), consisting of green crystals with a metallic lustre, is highly soluble in water, cytotoxic to various mammalian cells and also acts as a liver tumour promoter. In view of its industrial importance and possible exposure to human beings, MG poses a potential environmental health hazard. We have earlier reported the malignant transformation of Syrian hamster embryo (SHE) cells by MG. In this study, we have studied the effects of MG on cell cycle phase distribution of normal and MG transformed Syrian hamster embryo cells in asynchronous and synchronous cell population. DNA flow cytometric analysis indicated that culturing cells for 48 h in medium containing MG at different concentrations induced dose-dependent G2/M arrest in normal cells. Malignantly transformed cells showed no such dose-responsive accumulation of cells at the G2/M phase of the cell cycle in response to MG. Synchronization studies indicated that in the control, both in the presence and absence of MG, cells followed a normal cell cycle pattern up to 16 h. After 16 h in the absence of MG, cells continued a normal cell cycle, whereas in the presence of MG they accumulated at G2/M phase of the cell cycle. This pattern of accumulation of cells at the G2/M checkpoint control was not observed in either untreated or MG-treated transformed cells. The present study indicates efficient operation of G2/M checkpoint control in control SHE cells and its abrogation in transformed SHE cells. PMID- 10452829 TI - Selectivity in the distribution of glutamate receptors in neurons. PMID- 10452827 TI - Staurosporine-induced cell death in Tetrahymena thermophila has mixed characteristics of both apoptotic and autophagic degeneration. AB - Staurosporine blocks signal transduction associated with cell survival, proliferation and chemosensory behaviour in the ciliated protozoan, Tetrahymena thermophila. Staurosporine inhibits cell proliferation and in vivo protein phosphorylation induced by phorbol ester. It also reduces the in vitro phosphorylation of the PKC-specific substrate, myelin basic protein fragment 4 14. Our results show that cell death in the presence of staurosporine is associated with morphological and ultrastructural changes similar to both apoptosis and autophagic degeneration, but these in turn can be postponed or prevented by 8-bromo-cyclic GMP, protoporphyrin IX, hemin or actinomycin D, although phorbol ester and insulin were ineffective. The results support the notion that staurosporine-induced cell death is an active process, associated with and/or requiring de novo RNA synthesis. PMID- 10452830 TI - Detection of intranucleolar chromatin using an ultrastructural immunolabelling technique. AB - The intranucleolar chromatin has been detected by feeding HeLa cells with bromodeoxyuridine and then using an immunocytochemical technique for its detection at the ultrastructural level. Labelling was mainly observed on the dense fibrillar component (DFC), which surrounds the fibrillar centres (FC). Labelling could also be seen on the network within the granular component (GC) of the nucleoli. Ring shaped nucleoli with large FCs showed labelling, in addition, on the FCs. Nucleoli from spread preparations of cells treated with hypotonic salt solution for different periods revealed a nucleolonemal network, consisting of units comparable to rDNA transcriptional units in length and associated with tufts of fibrils and granules. Thread-like axes could be detected running through the units and in between two units. Chromatin could also be identified within these units by the immunolabelling technique. The observations strongly indicate that these units are identical with the rDNA transcriptional units. A dynamic nature of the nucleolar chromatin and nucleolar organization has been discussed. PMID- 10452831 TI - Effects of interleukin-6 on proliferation and proteoglycan metabolism in articular chondrocyte cultures. AB - Interleukin-6 (IL-6) levels are markedly increased in the synovial fluid of patients with rheumatoid arthritis or osteoarthritis. However, the effects of IL 6 on proliferation and proteoglycan metabolism in articular cartilage are not known. We demonstrated here the effects of human recombinant (hr) IL-6 on proliferation and proteoglycan metabolism in rabbit articular chondrocyte cultures. In vitro, these cells proliferated and produced abundant extracellular matrices. We found that 1-10 ng/ml of hrIL-6 inhibited proliferation to approximately 65% of control levels and suppressed colony formation induced by bFGF in soft agarose. The same concentration of hrIL-6 depressed proteoglycan synthesis to approximately 60% of control levels. Moreover, hrIL-6 significantly enhanced proteoglycan degradation induced by hrIL-1beta, although hrIL-6 alone did not affect proteoglycan degradation. These findings suggest that IL-6 is a negative regulator for chondrocyte proliferation and articular cartilage metabolism. PMID- 10452833 TI - Proliferation and collagen synthesis of human anterior cruciate ligament cells in vitro: effects of ascorbate-2-phosphate, dexamethasone and oxygen tension. AB - Clinical and experimental studies demonstrate that injured anterior cruciate ligaments (ACL) do not usually heal and that autografts used to repair the ACL rapidly weaken in the early period and take a long time to regain strength. The aim of this study was to develop an in vitro culture system in which environmental and biochemical factors influencing the proliferation and matrix synthesis of cells derived from human anterior cruciate ligaments can be studied. Primary cultures of human ACL cells were obtained by outgrowth from explants of normal ACL obtained at knee replacement for osteoarthritis in Dulbecco's minimum essential medium (DMEM). The effects of the additives 100 microm L-ascorbic acid 2-phosphate (Asc-2-P) and 10 n m dexamethasone (dex) on proliferation and collagen synthesis were assessed after 4, 8 and 12 days in culture. Ligament cells were grown at 0, 5, 10 and 21%p O(2)in the presence of 100 microm asc-2-P and 10 n m dex. DNA content was assessed using the Hoechst dye method and collagen synthesis by the incorporation of 5 mCi/ml [(3)H]proline after 3, 6 and 12 days in culture. At 21%p O(2), the presence of asc-2-P and dex induced significantly greater (P< 0.01, ANOVA) cell proliferation than with either additives alone. Greatest percentage collagen to total protein synthesis was observed when cells were grown in the presence of asc-2-P only. Least proliferation and percentage collagen to total protein synthesis was seen when both additives were omitted. Greatest cell proliferation was seen when cells were grown in 10%p O(2)and 5%p O(2)was associated with increased collagen synthesis. These results suggest that it is possible to study the effects of environmental and biochemical factors on human ACL healing in vitro. Our data suggest oxygen can influence certain biosynthetic activities of ACL cells. Low oxygen tensions lead to an increase in collagen production by ACL cells. However early responses to injury require extensive cell proliferation which may be activated at higher p O(2). Variation of p O(2)in ligaments during healing may therefore be an important modulator of successful repair. PMID- 10452832 TI - Altered phenotypic characteristics of T47d human breast cancer cells after prolonged growth in estrogen-deficient medium. AB - T47D human breast cancer cells were cultured in estrogen-deficient media for up to 32 months and the resulting cell line (L(hE(-))) exhibited unique phenotypic and genotypic characteristics. Compared to low passage (L) cells, the L(hE(-)) cells exhibited a significantly higher rate of proliferation, unique morphological features, advanced ploidy status and 5- to 10-fold higher levels of the estrogen receptor (ER) as determined by ligand binding and Western blot analysis. Sequence analysis of the DNA binding domain of the ER revealed a C-->A transversion which resulted in a H513N amino acid change. Treatment of L cells with 10 n m 17beta-estradiol (E2) resulted in a greater than two-fold increase in cell proliferation which was inhibited by tamoxifen, 4'-hydroxytamoxifen, ICI 164,384 and ICI 182,780. In contrast, 10 n m E2 caused a 70% decrease in growth of L(hE(-)) cells and this antimitogenic activity was blocked by ICI 164,384 and ICI 182,780 but not by tamoxifen or 4'-hydroxytamoxifen. L(hE(-)) cells were E2 responsive in transient transfection studies using a plasmid containing an estrogen-responsive element derived from the vitellogenin A2 gene promoter. These data show that the phenotypic and genotypic characteristics of L(hE(-)) T47D cells resemble those described for ER-negative cell lines stably transfected with the ER. PMID- 10452834 TI - Expression of the cell adhesion molecule E-cadherin by the human bone marrow stromal cells and its probable role in CD34(+) stem cell adhesion. AB - Bone marrow stroma is the physical basis of the haematopoietic microenvironment and regulates several key features of stem cell proliferation and differentiation. It plays a crucial role in maintaining haematopoietic homeostasis. Earlier studies have shown that this is achieved through interactions with the extracellular matrix and specific molecules called the cell adhesion molecules (CAMs). In this paper, we show that E-cadherin, a cell adhesion molecule which plays a crucial role in cell-cell aggregation during development, is also present in the bone marrow stroma. The expression of the CAM can also be demonstrated on a subset of CD34(+)stem cells. Stromal expression of E-cadherin is decreased when treated with lymphokine mixture, phytohaemagglutinin treated-leukocyte-conditioned medium (PHA-LCM). This is the reverse of ICAM-I expression, which increases with PHA-LCM treatment. E-cadherin shows homotypic and homophilic interaction and its presence on a subset of CD34(+)cells leads to speculation on whether this CAM has a role in adherence of primitive stem cells to the marrow stroma. PMID- 10452835 TI - Binding contribution between synaptic vesicle membrane and plasma membrane proteins in neurons: an AFM study. AB - The final step in the exocytotic process is the docking and fusion of membrane bound secretory vesicles at the cell plasma membrane. This docking and fusion is brought about by several participating vesicle membrane, plasma membrane and soluble cytosolic proteins. A clear understanding of the interactions between these participating proteins giving rise to vesicle docking and fusion is essential. In this study, the binding force profiles between synaptic vesicle membrane and plasma membrane proteins have been examined for the first time using the atomic force microscope. Binding force contributions of a synaptic vesicle membrane protein VAMP1, and the plasma membrane proteins SNAP-25 and syntaxin, are also implicated from these studies. Our study suggests that these three proteins are the major, if not the only contributors to the interactive binding force that exist between the two membranes. PMID- 10452836 TI - The native membrane fusion machinery in cells. AB - Recombinant SNAREs have been demonstrated as the minimal membrane fusion machinery. The participation of additional proteins in the regulation of membrane fusion has been suggested. In this study we provide nanometer-resolution images of native NSF oligomers and SNARE complexes isolated from neurons and the pancreas. Our study reveals the presence of new coiled rod-like structures in association with the SNARE complex only in neuronal tissue. Neuronal SNAREs were found coiled and super-coiled with these structures. The existence of NSF as pentamers in its native state is also demonstrated. The extent of coiling and super-coiling of SNAREs may regulate the potency and efficacy of membrane fusion in cells. PMID- 10452837 TI - Apoptosis in human lung epithelial cells: triggering by paraquat and modulation by antioxidants. AB - Recent results have shown that apoptosis is an important feature of the normal and injured lung epithelium, but little conclusive evidence is available about the exact intracellular mechanisms involved. In this work, we studied apoptotic cell death in the established human lung epithelial cell line, A549, by evaluating the ability of the pulmonary toxin, paraquat (1,1'-dimethyl-4, 4' bipyridylium dichloride), to act as a trigger, and assessing the ability of ascorbic acid and N-acetyl-cysteine (NAC) to modulate the process. The analysis of nuclear and cellular morphology along with TUNEL staining showed that paraquat is an inducer of apoptosis. A549 cells incubated with sublethal doses of paraquat for up to 24 h showed no apoptotic features but, their following incubation in paraquat-free medium resulted in a time-dependent appearance of apoptosis. The antioxidants, ascorbic acid and NAC, proved effective in reducing paraquat induced apoptosis, and therefore were seen as protective agents. Finally, we propose an experimental model for investigating some of the key steps in the apoptotic programme in alveolar cells. PMID- 10452838 TI - Growth of MDA-MB-231 cell line: different effects of TGF-beta(1), EGF and estradiol depending on the length of exposure. AB - The human cell line MDA-MB-231 is a prototype for the study of hormone independent breast cancer. Modification of cell growth behaviour has been observed after treating these cells with growth factors. EGF is a typical stimulatory growth factor for many cell types, whereas transforming growth factor beta(1)(TGF-beta(1)) acts with inhibitory character. Here we observed cell growth inhibition after EGF as well as after TGF-beta(1)treatments. Nevertheless, in the 42-h experiments, EGF-treated cultures grew before (18 hours) respect to the TGF beta(1)and E(2)-treated cultures (24 h), and in the 11-day experiments, EGF treated cultures started growing (7 days) after TGF-beta(1)-treated cultures (5 days). Estradiol inhibited the proliferation of these cells only after several days of treatment. PMID- 10452839 TI - Participation of two isoforms of C/EBPbeta transcription factor in the acute phase regulation of the rat haptoglobin gene. AB - Previous analyses of the mechanism of the transcriptional induction of the rat haptoglobin (Hp) gene during acute-phase (AP)-reaction have revealed the involvement of several trans-acting nucleoproteins (NPs) in controlling this process. In this study, by using antibodies against C/EBPbeta factor in Western immunoblot assay, we found that rat liver trans-acting NPs p35 and p20 are two characteristic C/EBPbeta isoforms whose expression is induced under AP conditions. DNA-binding assays identified the binding sites for these two C/EBPbeta proteins in the functionally defined elements A and C of the rat Hp gene and also revealed that they have specific binding affinity towards these elements. Under non-induced conditions, p35 was the only C/EBPbeta binding factor; however, upon AP-conditions both, 35 kDa- and 20 kDa-C/EBPbeta binding activities were significantly induced suggesting that these interactions are necessary for the activation of the Hp gene. By in vitro phosphorylation assay and selective proteolysis, we also present evidence that p35 requires phosphorylation for its DNA binding ability. Thus, we conclude that increase in binding of C/EBPbeta isoforms during AP-reaction occurs through their upregulation and structural modification. PMID- 10452840 TI - A study of the effect of theophylline on the anti-inflammatory potency of dexamethasone using alpha-aminoisobutyric acid uptake in rat skin fibroblasts: a possible mechanism through the glucocorticoid receptor. AB - The action of theophylline on the uptake of alpha-aminoisobutyric acid (AIB; an indicator of anti-inflammatory potency) stimulated by the glucocorticoid, dexamethasone, in cultured rat fibroblast monolayers was evaluated. Theophylline alone (0.1 m m) did not show significant activity (3314+/-27 cpm) compared with the baseline level (3186+/-130 cpm), but in the presence of 10 n m dexamethasone the stimulation of AIB uptake was increased to 5263+/-100 cpm, approximately to the same extent as with 100 n m dexamethasone alone (5397+/-28 cpm). Activation of glucocorticoid-receptor complexes in rat fibroblast cytosol was studied by assessing the extent of their binding to DNA-cellulose. Activated and non activated forms of glucocorticoid-receptor complexes were analysed by DEAE Sephadex chromatography. Theophylline (1 m m) was found to have a direct effect (0 degrees C), similar to that of heat (25 degrees C) on DNA-cellulose binding, that is approximately 64.5% and 68.7%, respectively, thus indicating that theophylline promotes activation of glucocorticoid receptors at low temperature. The effect of theophylline on the stimulation of AIB uptake by dexamethasone when considered in the light of its activation of GR receptors in the fibroblast cytosol indicates that this effect may be mediated by GR activation. PMID- 10452841 TI - Effects of cold acclimation on the activity levels of creatine kinase, lactate dehydrogenase and lactate dehydrogenase isoenzymes in various tissues of the rat. AB - The effects of cold acclimation on the activity levels of creatine kinase, lactate dehydrogenase and lactate dehydrogenase isoenzymes in various tissues/ organs of the rat (Rattus norvegicus) were investigated. Male Sprague-Dawley rats were divided into two groups. One group was housed at 4+/-1 degrees C (experimental group) and the other at 24+/-1 degrees C (control group) for six months. The rats were housed in single cages and had access to food and water ad libitum. The tissues/organs investigated were heart, liver, lung, kidney, gastrocnemius muscle and interscapular brown adipose tissue as well as serum. With the exception of lung, (which showed a decrease of 24%) total creatine kinase activity levels were significantly increased (P< 0.05) in all the tissues/organs investigated (17-51%) as well as serum (34%), in cold acclimated animals. Cold acclimation also resulted in significantly increased (P< 0.05) activity levels of lactate dehydrogenase in all the tissues/organs investigated (14-24%) as well as serum (35%). Cold exposure resulted in an increase of the activity levels of all the detectable isoenzymes of lactate dehydrogenase, although not always significant, in all the tissues/organs investigated as well as serum. The M(4)tetramer of lactate dehydrogenase was the only detectable isoenzyme in serum. PMID- 10452842 TI - Resumption of rapid proliferation from lag phase in cultures of Saccharomyces cerevisiae in poor nutrient conditions. Effect of surface and intracellular signalling mechanisms. AB - Saccharomyces cerevisiae was inoculated into a dilute synthetic minimal medium with glycerol as the carbon source. The number of live cells in the cultures was determined by colony counts on agar plates. Untreated control cells had doubled in number about once at the end of the first week and had gone through eight doublings by the end of the second week. Addition of either 8-bromo-cyclic guanosine monophosphate (8-bromo-cGMP) or human recombinant insulin, made the cells go through 12 and 10 doublings, respectively, by the end of the first week. In contrast, 8-bromo-cyclic adenosine monophosphate (8-bromo-cAMP) had only slight stimulating effects on cell multiplication, but if it was combined with phorbol-12-myristate-13-acetate (PMA) the cells went through about 12 doublings during the first week. Addition of LY 83583, an inhibitor of soluble guanylate cyclase, prevented cell proliferation. Further addition of 8-bromo-cGMP bypassed this inhibition. Singly, bradykinin or PMA did not affect cell multiplication. However, when these two compounds were combined, the cells went through about 10 doublings during the first week. Neither bradykinin, nor PMA had any releasing effect on the inhibition of LY 83583. These results indicate the existence of several routes leading to cell proliferation in wildtype S. cerevisiae cells. PMID- 10452843 TI - Oocyte apoptosis: like sand through an hourglass. AB - Although the study of germ cell death is arguably still in its infancy as a field, several recent breakthroughs have provided the fodder for a story, replete with episodes of apparent mass cellular suicide if not murder, that will undoubtedly serve as a research base for many laboratories over the next several years. Death is known to strike the male and female germlines with roughly equal intensity, but the innate feature of male germ cells being self-renewing while those of the female are not places the death of oocytes in a completely different light. Indeed, the functional life span of the female gonads is defined in most species, including humans, by the size and rate of depletion of the precious endowment of oocytes enclosed within follicles in the ovaries at birth. This continuous loss of oocytes throughout life, referred to by many as the female biological clock, appears to be driven by a genetic program of cell death that is composed of players and pathways conserved from worms to humans. It is on this genetic pathway, and the role of its constituent molecules in regulating female germ cell fate, that this review will focus. Emphasis will be placed on those studies using genetic-null or transgenic models to explore the functional requirement of proteins, such as Bcl-2 family members, Apaf-1, and caspases in vertebrates to CED-9, CED-4, and CED-3 in Caenorhabditis elegans, in oocyte survival and death. Furthermore, hypotheses regarding the potential impact of translating what is now known of the oocyte death pathway into new approaches for the clinical diagnosis and management of female infertility and the menopause will be offered as a means to stimulate further research in this new and exciting field. PMID- 10452844 TI - Two forms of vitellogenin, yielding two distinct lipovitellins, play different roles during oocyte maturation and early development of barfin flounder, Verasper moseri, a marine teleost that spawns pelagic eggs. AB - Two forms of vitellogenin (Vg), Vg A and Vg B, were identified in serum from estrogen-treated barfin flounder (Verasper moseri). Structural changes of lipovitellins (Lvs) derived from the two Vgs were examined during vitellogenesis and oocyte maturation. Two Lvs, vLv A and vLv B, were identified electrophoretically and immunologically in postvitellogenic oocytes. Each appeared to be composed of distinct heavy chains (vLvH A, M(r) 107,000, and vLvH B, M(r) 94,000) and light chains (vLvL A, M(r) 30,000, and vLvL B, M(r) 28,000) when analyzed by SDS-PAGE. Results from N-terminal amino acid sequencing and Western blotting using antisera to vLvH A and vLvH B verified that there are two Vg polypeptides in serum from estrogen-treated fish, Vg A (M(r) 168,000) and Vg B (M(r) 175,000), which give rise to vLvH A-vLvL A and vLvH B-vLvL B, respectively. N-terminal sequencing revealed two sequences for both phosvitin and beta' component, supporting the concept of duality for all three classes of Vg-derived yolk proteins. During oocyte maturation, native dimeric vLv B was dissociated into a native M(r) 170,000 monomer (oLv B). Meanwhile, vLv A was extensively cleaved including complete degradation of vLvH A into free amino acids. We propose that the quantitative ratio of vLv A to vLv B in postvitellogenic oocytes regulates the buoyancy of the spawned pelagic eggs by controlling availability of free amino acids which function as osmotic effectors during oocyte hydration. The vLv A/vLv B ratio likely also controls the proportional availability of different types of nutrients, free amino acids versus Lv, for use during embryonic development. PMID- 10452845 TI - Discrete enhancer elements mediate selective responsiveness of enhancer of split complex genes to common transcriptional activators. AB - In Drosophila, genes of the Enhancer of split Complex [E(spl)-C] are important components of the Notch (N) cell-cell signaling pathway, which is utilized in imaginal discs to effect a series of cell fate decisions during adult peripheral nervous system development. Seven genes in the complex encode basic helix-loop helix (bHLH) transcriptional repressors, while 4 others encode members of the Bearded family of small proteins. A striking diversity is observed in the imaginal disc expression patterns of the various E(spl)-C genes, suggestive of a diversity of function, but the mechanistic basis of this variety has not been elucidated. Here we present strong evidence from promoter-reporter transgene experiments that regulation at the transcriptional level is primarily responsible. Certain E(spl)-C genes were known previously to be direct targets of transcriptional activation both by the N-signal-dependent activator Suppressor of Hairless [Su(H)] and by the proneural bHLH proteins achaete and scute. Our extensive sequence analysis of the promoter-proximal upstream regions of 12 transcription units in the E(spl)-C reveals that such dual transcriptional activation is likely to be the rule for at least 10 of the 12 genes. We next show that the very different wing imaginal disc expression patterns of E(spl)m4 and E(spl)mgamma are a property of small (200-300 bp), evolutionarily conserved transcriptional enhancer elements, which can confer these distinct patterns on a heterologous promoter despite their considerable structural similarity [each having three Su(H) and two proneural protein binding sites]. We also demonstrate that the characteristic inactivity of the E(spl)mgamma enhancer in the notum and margin territories of the wing disc can be overcome by elevated activity of the N receptor. We conclude that the distinctive expression patterns of E(spl)-C genes in imaginal tissues depend to a significant degree on the capacity of their transcriptional cis-regulatory apparatus to respond selectively to direct proneural- and Su(H)-mediated activation, often in only a subset of the territories and cells in which these modes of regulation are operative. PMID- 10452846 TI - Pattern formation in the absence of cell proliferation: tissue-specific regulation of cell cycle progression by string (stg) during Drosophila eye development. AB - During Drosophila eye development, the posterior-to-anterior movement of the morphogenetic furrow coordinates cell cycle progression with the early events of pattern formation. The cdc25 phosphatase string (stg) has been proposed to contribute to the synchronization of retinal precursors anterior to the furrow by driving cells in G(2) through mitosis and into a subsequent G(1). Genetic and molecular analysis of Drop (Dr) mutations suggests that they represent novel cis regulatory alleles of stg that inactivate expression in eye. Retinal precursors anterior to the furrow lacking stg arrest in G(2) and fail to enter mitosis, while cells within the furrow accumulate high levels of cyclins A and B. Although G(2)-arrested cells initiate normal pattern formation, the absence of stg results in retinal patterning defects due to the recruitment of extra photoreceptor cells. These results demonstrate a requirement for stg in cell cycle regulation and cell fate determination during eye development. PMID- 10452847 TI - A paraxial exclusion zone creates patterned cranial neural crest cell outgrowth adjacent to rhombomeres 3 and 5. AB - Cranial neural crest cell migration is patterned, with neural crest cell-free zones adjacent to rhombomere (R) 3 and R5. These zones have been suggested to result from death of premigratory neural crest cells via upregulation of BMP-4 and Msx-2 in R3 and R5, consequent to R2-, R4-, and R6-derived signals. We reinvestigated this model and found that cell death detected by acridine orange staining in avian embryos varied widely numerically and in pattern, but with a tendency for an elevated zone centered at the R2/3 boundary. In situ hybridization of BMP-4 mRNA resolved to centers at R3 and R5 but Msx-2 resolved to the R2/3 border with only a faint smear from R5 to R6. Outgrowth of neural crest cells was less in isolated R3 cultures than in R1+2, R2, and R4 cultures, but R3 showed neither a decrease in outgrowth of neural crest cells nor an increase in cell death when cocultured with R1+2, R2, or R4. In addition, in serum-free culture, exogenous BMP-4 strikingly reduced neural crest cell outgrowth from R1+2 and R4 as well as R3. Thus we cannot confirm the role of intraneural cell death in patterning rhombomeric neural crest outgrowth. However, grafting quail R2 or R4 adjacent to the chick hindbrain demonstrated a neural crest cell exclusion zone next to R3 and R5. We suggest that one important pattern determinant for rhombomeric neural crest cell migration involves the microenvironment next to the neural tube. PMID- 10452848 TI - Interference with brachyury function inhibits convergent extension, causes apoptosis, and reveals separate requirements in the FGF and activin signalling pathways. AB - Brachyury plays a key role in mesoderm formation during vertebrate development. Absence of the gene results in loss of posterior mesoderm and failure of the notochord to differentiate, while misexpression of Brachyury in the prospective ectoderm of Xenopus results in ectopic mesoderm formation. Brachyury is therefore both necessary and sufficient for posterior mesoderm formation. Here we present a detailed cellular and molecular analysis of the consequences of inhibiting Brachyury function during Xenopus development. Our results show that Brachyury is required for the convergent extension movements of gastrulation, for mesoderm differentiation in response to FGF, and for the survival of posterior mesodermal cells in both Xenopus and mouse. PMID- 10452849 TI - Inhibition of neural crest migration in Xenopus using antisense slug RNA. AB - Based primarily on studies in the chick, it has been assumed that the zinc finger transcription factor Slug is required for neural crest migration. In the mouse, however, Slug is not expressed in the premigratory neural crest, which forms normally in Slug -/- animals. To study the role of Slug in Xenopus laevis, we used the injection of XSlug antisense RNA and tissue transplantation. Injection of Slug antisense RNA did not suppress the early expression of the related gene XSnail, but led to reduced expression of both XSlug and XSnail in later stage embryos, whereas the expression of another neural crest marker, XTwist, was not affected. Down-regulation of XSlug and XSnail was associated with the inhibition of neural crest cell migration and the reduction or loss of many neural crest derivatives. In particular, the formation of rostral cartilages was often highly aberrant, whereas the posterior cartilages were less frequently affected. The effects of Slug antisense RNA on neural crest migration and cartilage formation were rescued by the injection of either XSlug or XSnail mRNA. These studies indicate that XSlug is required for neural crest migration, that XSlug and XSnail may be functionally redundant, and that both genes are required to maintain each other's expression in the neural crest development of xenopus laevis. PMID- 10452850 TI - The 82F late puff contains the L82 gene, an essential member of a novel gene family. AB - Metamorphosis in Drosophila results from a hierarchy of ecdysone-induced gene expression initiated at the end of the third larval instar. A now classical model of this hierarchy was proposed based on observations of the activity of polytene chromosome "puffs" which distinguished "early" puffs as those directly induced by ecdysone and "late" puffs as those which become active as a secondary response to the hormone. We report here the isolation and characterization of the L82 gene corresponding to the extensively characterized late puff at 82F. L82 is a complex gene that spans at least 50 kb of genomic DNA, produces at least seven different nested mRNAs, and has homology to a novel gene family. In contrast to most previously characterized puff genes, the broad developmental expression pattern of L82 suggests that it is controlled by both ecdysone-dependent and ecdysone independent regulatory mechanisms. L82 mutations were identified by transgene rescue of developmental delay and eclosion lethal phenotypes. PMID- 10452851 TI - Colinear and segmental expression of amphioxus Hox genes. AB - The cephalochordate amphioxus has a single Hox gene cluster. Here we describe the genomic organization of four adjacent amphioxus genes, AmphiHox-1 to AmphiHox-4, together with analysis of their spatiotemporal expression patterns. We demonstrate that these genes obey temporal colinearity and that three of the genes also obey spatial colinearity in the developing neural tube. AmphiHox-1, AmphiHox-3, and AmphiHox-4 show segmental modulation of their expression levels, a two-segment phasing of spatial colinearity, and, at least for AmphiHox-4, asymmetrical expression. AmphiHox-2 is unlike other amphioxus Hox genes: it does not obey spatial colinearity and it has no positional expression in the neural tube. AmphiHox-2 is expressed in the preoral pit of larvae, from which the homologue of the anterior pituitary develops. We suggest that the ancestral role of chordate Hox genes was primarily in the neural tube and that chordate Hox genes can functionally diverge in a manner analogous to that of Drosophila ftz or zen. PMID- 10452852 TI - Members of the bcl-2 and caspase families regulate nuclear degeneration during chick lens fibre differentiation. AB - The optical clarity of the lens is ensured by the programmed removal of nuclei and other organelles from the lens fibre cells during development. The morphology of the degenerating nuclei is similar to that observed during apoptosis and is accompanied by DNA fragmentation. Proteins encoded by the bcl-2 proto-oncogene family are important in either promoting or inhibiting apoptosis, and caspases are involved in downstream proteolytic events. Here, the expression of bcl-2 family members (bcl-2, bax, bad, and bcl-x(s/l)) and caspases-1, -2, -3, -4, and 6 was investigated through a range of stages of chick lens development using immunocytochemistry, Western blotting, and affinity labelling for caspases using biotinylated caspase inhibitors. Using differentiating lens epithelial cell cultures, it was demonstrated that the addition to cultures of synthetic peptide inhibitors of caspases -1, -2, -4, -6, and -9 brought about a 50-70% reduction in the number of degenerating nuclei per unit area of culture, as assessed by image analysis. These effects were comparable to those seen when general inhibitors of caspases were added to cultures. On the other hand, inhibitors of caspases-3 and 8 were not effective in significantly reducing the number of TUNEL-labelled nuclei. Expression of the caspase substrates poly(ADP-ribose) polymerase (PARP) and the 45-kDa subunit of DNA fragmentation factor (DFF 45) was also observed in the developing lens. Western blots of cultures to which caspase inhibitors were added revealed alterations in the PARP cleavage pattern, but not in that of DFF. These results demonstrate a role for members of the bcl-2 family and caspases in the degeneration of lens fibre cell nuclei during chick secondary lens fibre development and support the proposal that this process has many characteristics in common with apoptosis. PMID- 10452853 TI - The type II activin receptors are essential for egg cylinder growth, gastrulation, and rostral head development in mice. AB - The type II activin receptors, ActRIIA and ActRIIB, have been shown to play critical roles in axial patterning and organ development in mice. To investigate whether their function is required for mesoderm formation and gastrulation as implicated in Xenopus studies, we generated mice carrying both receptor mutations by interbreeding the ActRIIA and ActRIIB knockout mutants. We found that embryos homozygous for both receptor mutations were growth arrested at the egg cylinder stage and did not form mesoderm. Further analyses revealed that ActRIIA(-/ )ActRIIB(+/-) and about 15% of the ActRIIA(-/-) embryos failed to form an elongated primitive streak, resulting in severe disruption of mesoderm formation in the embryo proper. Interestingly, we observed similar gastrulation defects in ActRIIA(-/-)nodal(+/-) double mutants, which, if they developed beyond the gastrulation stage, displayed rostral head defects and cyclopia. These results provide genetic evidence that type II activin receptors are required for egg cylinder growth, primitive streak formation, and rostral head development in mice. PMID- 10452854 TI - A role for xGCNF in midbrain-hindbrain patterning in Xenopus laevis. AB - Cells in the presumptive neural ectoderm of Xenopus are committed to neural fate through a process called neural induction, which may involve proteins that antagonize BMP signaling pathways. To identify genes that are induced by the BMP antagonists and that may be involved in subsequent neural patterning, we used a suppression PCR-based subtraction screen. Here we investigate the prospective activities and functions of one of the genes, a nuclear orphan receptor previously described as xGCNF. In animal cap assays, xGCNF synergizes with ectopic chordin to induce the midbrain-hindbrain marker engrailed-2 (En-2). In Keller explants, which rely on endogenous factors for neural induction, similar increases in En-2 are observed. Expression in embryos of a dominant interfering form of xGCNF reduces the expression of endogenous En-2 and Krox-20. These gain of-function and prospective loss-of-function experiments, taken with the observation that xGCNF is expressed in the early neural plate and is elevated in the prospective midbrain-hindbrain region, which subsequently expresses En-2, suggest that xGCNF may play a role in regulating En-2 and thus midbrain-hindbrain identity. PMID- 10452855 TI - Retinoic acid signaling is necessary for the development of the organ of Corti. AB - The cellular mosaic of the mammalian organ of Corti represents one of the most highly ordered structures in any vertebrate system. A single row of inner hair cells and three or four rows of outer hair cells extend along the basal-to-apical axis of the cochlea. The factors that play a role in the development of specific cell types within the cochlea are largely unknown; however, the results of previous studies have strongly suggested that retinoic acid plays a role in the development of cells as hair cells. To determine whether cochlear progenitor cells can respond directly to retinoic acid, the expression patterns for each of the RAR and RXR receptors within the embryonic cochlear duct were determined by in situ hybridization. Results indicate that RARalpha, RXRalpha, and RXRgamma are initially expressed throughout the cochlear duct. As development continues, the expression of each receptor becomes more intense in cells that will develop as hair cells. At the same time, receptor expression is down-regulated in cells that will develop as nonsensory cell types. To determine the effects of retinoic acid signaling during the development of the organ of Corti, activation of retinoid receptors was blocked in cultures of the embryonic cochlea through receptor specific antagonism or inhibition of retinoic acid synthesis. Results indicate that inhibition of retinoic acid signaling induces a significant decrease in the number of cells that develop as hair cells and a disruption in the development of the organ of Corti. These results demonstrate that cells within the developing cochlea can respond to retinoic acid and that signaling by retinoic acid is necessary for the normal development of the organ of Corti. PMID- 10452856 TI - Reversible inactivation of cell-type-specific regulatory and structural genes in migrating isolated striated muscle cells of jellyfish. AB - We have investigated, by RT-PCR and in situ hybridization, expression of genes encoding regulatory and structural proteins in migrating mononucleated striated muscle cells of the medusa Podocoryne carnea. Expression of the three homeobox genes Otx, Cnox1-Pc, and Cnox3-Pc; a specific splice variant of the myosin heavy chain gene (Myo1); and a tropomyosin (Tpm2) is stable in isolated and cultured striated muscle tissue. When grafted onto cell-free extracellular matrix (ECM), muscle cells of the tissue fragments leave their native ECM and migrate as a coherent tissue onto a host ECM until a stretched cell monolayer is formed. Shortly after the first cells of the grafted isolate have made contact with the host ECM, Otx and Cnox1-Pc expression is completely turned off in all cells of the graft, including those still adhering to their native ECM. Myo1 message disappears with a delay while the expression level of Tpm2 is strongly reduced. However, expression of the homeobox gene Cnox3-Pc, a msh-like gene, and of the ubiquitously expressed elongation factor 1 alpha is not affected by the migration process. All genes are reexpressed after 12-24 h, once migration of the cells has ceased. Our results demonstrate that the first few migrating cells induce a change in gene expression which is rapidly communicated throughout the entire tissue. Furthermore, we showed that commitment of striated muscle cells remains stable despite the transient inactivation of cell-type-specific regulatory and structural genes. PMID- 10452857 TI - The mouse Cer1 (Cerberus related or homologue) gene is not required for anterior pattern formation. AB - Cer1 is the mouse homologue of the Xenopus Cerberus gene whose product is able to induce development of head structures during embryonic development. The Cer1 protein is a member of the cysteine knot superfamily and is expressed in anterior regions of the mouse gastrula. A segmental pattern of expression with nascent and newly formed somites is also seen. This suggests an additional role in development of the axial skeleton, musculature, or peripheral nervous system. Xenopus animal cap assays and mouse germ-layer explant recombination experiments indicate that the mouse protein can act as a patterning molecule for anterior development in Xenopus, including induction of Otx2 expression, and suggest it may have a similar role in mouse development. However, we present here genetic data that demonstrate that Cer1 is not necessary for anterior patterning, Otx2 expression, somite formation, or even normal mouse morphogenesis. PMID- 10452858 TI - Transgene expression in zebrafish: A comparison of retroviral-vector and DNA injection approaches. AB - To assess alternative methods for introducing expressing transgenes into the germ line of zebrafish, transgenic fish that express a nuclear-targeted, enhanced, green fluorescent protein (eGFP) gene were produced using both pseudotyped retroviral vector infection and DNA microinjection of embryos. Germ-line transgenic founders were identified and the embryonic progeny of these founders were evaluated for the extent and pattern of eGFP expression. To compare the two modes of transgenesis, both vectors used the Xenopus translational elongation factor 1-alpha enhancer/promoter regulatory cassette. Several transgenic founder fish which transferred eGFP expression to their progeny were identified. The gene expression patterns are described and compared for the two modes of gene transfer. Transient expression of eGFP was detected 1 day after introducing the transgenes via either DNA microinjection or retroviral vector infection. In both cases of gene transfer, transgenic females produced eGFP-positive progeny even before the zygotic genome was turned on. Therefore, GFP was being provided by the oocyte before fertilization. A transgenic female revealed eGFP expression in her ovarian follicles. The qualitative patterns of gene expression in the transgenic progeny embryos after zygotic induction of gene expression were similar and independent of the mode of transgenesis. The appearance of newly synthesized GFP is detectable within 5-7 h after fertilization. The variability of the extent of eGFP expression from transgenic founder to transgenic founder was wider for the DNA-injection transgenics than for the retroviral vector-produced transgenics. The ability to provide expressing germ-line transgenic progeny via retroviral vector infection provides both an alternative mode of transgenesis for zebrafish work and a possible means of easily assessing the insertional mutagenesis frequency of retroviral vector infection of zebrafish embryos. However, because of the transfer of GFP from oocyte to embryo, the stability of GFP may create problems of analysis in embryos which develop as quickly as those of zebrafish. PMID- 10452860 TI - Joel Jay Roslyn, M.D. (1951-1999). PMID- 10452859 TI - An E box comprises a positional sensor for regional differences in skeletal muscle gene expression and methylation. AB - To dissect the molecular mechanisms conferring positional information in skeletal muscles, we characterized the control elements responsible for the positionally restricted expression patterns of a muscle-specific transgene reporter, driven by regulatory sequences from the MLC1/3 locus. These sequences have previously been shown to generate graded transgene expression in the segmented axial muscles and their myotomal precursors, fortuitously marking their positional address. An evolutionarily conserved E box in the MLC enhancer core, not recognized by MyoD, is a target for a nuclear protein complex, present in a variety of tissues, which includes Hox proteins and Zbu1, a DNA-binding member of the SW12/SNF2 gene family. Mutation of this E box in the MLC enhancer has only a modest positive effect on linked CAT gene expression in transfected muscle cells, but when introduced into transgenic mice the same mutation elevates CAT transgene expression in skeletal muscles, specifically releasing the rostral restriction on MLC-CAT transgene expression in the segmented axial musculature. Increased transgene activity resulting from the E box mutation in the MLC enhancer correlates with reduced DNA methylation of the distal transgenic MLC1 promoter as well as in the enhancer itself. These results identify an E box and the proteins that bind to it as a positional sensor responsible for regional differences in axial skeletal muscle gene expression and accessibility. PMID- 10452861 TI - Reperfusion injury of the liver: role of mitochondria and protection by glutathione ester. AB - BACKGROUND: Reperfusion injury of the liver is characterized by intravascular oxidative stress and GSH consumption. Whether mitochondria contribute to hepatocellular damage has never been elucidated. Therefore, we assessed mitochondrial function and redox state during reperfusion and the effect of glutathione monoethyl ester (GSHE) administration, which may replenish the GSH pool. MATERIALS AND METHODS: Rats were subjected to partial hepatic ischemia (90 min) followed by reperfusion. Mitochondrial function was assessed in vivo and in vitro by the KICA breath test and the ATP synthase activity. Just prior to the start of reperfusion, rats received 5 mmol/kg of GSHE or saline iv. ALT, total and oxidized (GSSG) glutathione, GSHE, and CYS were measured in plasma and liver. GSH, GSSG, malondialdehyde (MDA), and carbonyl proteins were measured in mitochondria. The extent of necrosis was also estimated. Sham-operated rats served as controls. RESULTS: Reperfusion markedly increased ALT (>1500 U/L) and doubled the liver content of MDA and carbonyl proteins. Mitochondrial GSH decreased approximately 30%, without increase of GSSG. The in vivo KICA breath test was not significantly impaired by reperfusion. In contrast, both KICA decarboxylation and ATP synthase activity were both reduced by approximately 50% in mitochondria isolated from reperfused livers. GSHE administration significantly decreased ALT ( approximately 40%), protected ATP synthase activity, and reduced the extent of necrosis. Compared to controls, plasma GSHE and plasma GSH at 1 h were lower in rats subjected to ischemia. GSHE was higher in reperfused lobes than in continuously perfused ones and the concentration of GSH was significantly higher in ischemic liver than in untreated animals, indicating that the uptake of GSHE is increased in postischemic liver. GSHE prevented the reperfusion-associated increase of oxidized products in liver and mitochondria. CONCLUSIONS: Reperfusion of ischemic liver is associated with oxidative modifications and functional impairment of mitochondria. GSHE protects against reperfusion injury, possibly by providing intra- and extracellular GSH. PMID- 10452862 TI - Optimizing fluorescent labeling of endothelial cells for tracking during long term studies of autologous transplantation. AB - BACKGROUND: The fluorescent marker PKH26 has been demonstrated to be useful for the tracking of endothelial cells in short-term studies; however, the optimal labeling conditions for long-term implants have not been determined. This study was designed to evaluate the effects of PKH26 on endothelial cell proliferation and to identify labeling conditions that would yield the greatest fluorescence over time without adversely affecting cell viability. MATERIALS AND METHODS: Canine jugular vein endothelial cells (CJVECs) were labeled with 0. 04 microM PKH26. Proliferation of labeled and control cells was assessed for 8 consecutive days by [(3)H]thymidine uptake. In a second experiment, CJVECs were labeled at concentrations of 0, 5, 8, 10, and 20 micromol/L. Cells were maintained in culture for 60 days. The fluorescence intensity of each cell population was measured using two techniques. At baseline and at 60 days, fluorescence was measured using a fluorescence-activated cell sorter. On days 14, 28, 45, and 60 fluorescence was measured by constructing gray-scale histograms from photomicrographs taken of each flask under rhodamine illumination. Mean viable cell number for each concentration was determined after 60 days. RESULTS: In the first experiment, PKH26-labeled and unlabeled CJVECs demonstrated nearly identical growth curves, suggesting that PKH26 had no adverse effect on proliferation. In the second experiment, after 60 days, the 10 and 20 microM groups displayed greater fluorescence by histogram than the 0, 5, or 8 microM groups; however, they were not significantly different from each other (mean intensity 8.2 vs 9.1, P > 0.05, Student-Newman-Keuls test for multiple comparisons). Over 60 days, the cells labeled with 20 microM PKH26 experienced the only significant decrease in viable cells compared to the unlabeled group (5.5 x 10(5) vs 9.6 x 10(5) cells/flask, P < 0.05). Importantly, we observed no significant differences in cell number between the 10 microM group and the lower concentrations compared to the unlabeled cells (P > 0.05). CONCLUSIONS: We conclude that a concentration of 10 microM PKH26 provides the optimal labeling condition for endothelial cells when long-term tracking is desired. PMID- 10452863 TI - The relative contributions of interpersonal and specific clinical skills to the perception of global clinical competence. AB - BACKGROUND: The objective structured clinical examination (OSCE), an established instrument for evaluating resident competence, was used to test the hypothesis that faculty assessment of clinical competence in residents at various levels of training may be influenced more by general skills as a physician and less by competency in the actual skills being specifically tested. In this study, advantage was taken of the anticipated observation that general surgery residents did not demonstrate improvement in their ability to perform a focused neurological assessment over time. METHODS: An OSCE, which was administered to 56 general surgery residents at all levels of training, included the assessment of a specific clinical neurosurgical problem (sciatica). Univariate and multivariate analyses were used to evaluate the relationship between the global faculty judgment of competent or noncompetent and the other performance measures that were applied. RESULTS: At different levels of training, there was no observed difference in the specific skills being tested; nevertheless, junior and senior residents were more likely than incoming interns to be judged "competent" and received better evaluations of how well they introduced themselves to the patient. The competence judgment correlated significantly with all of the other performance measures, including the skills being tested. CONCLUSIONS: The perception of competence is not solely dependent upon the particular skills under scrutiny. General competence does not guarantee competence in each specific skill set of a medical specialty. PMID- 10452864 TI - Increased rat cardiac allograft survival by the glycosaminoglycan pentosan polysulfate. AB - BACKGROUND: Modulation of the inflammatory response has proven to be of benefit in salvaging cardiac allografts at risk of irreversible injury. Pentosan polysulfate (PPS), like heparin, is a negatively charged sulfated glycosaminoglycan (GAG) that possesses anti-inflammatory properties including the ability to inhibit activation of the complement system. This study was conducted to determine the potential of PPS to prolong allograft survival in an experimental model of cardiac transplantation. MATERIALS AND METHODS: A heterotopic cardiac transplant was performed by implanting the heart from fetal Brown Norway rats into the ear pinnae of adult Lewis rats. Vehicle (saline) or PPS (30 mg/kg) was administered subcutaneously immediately after transplantation and daily thereafter (n = 6 in each group). Another GAG, heparin, was also analyzed to determine the effect of anticoagulation on transplant survival (n = 6). RESULTS: Treatment with PPS significantly (P < 0. 05) increased allograft survival time as compared to vehicle-treated animals (8.0 +/- 0.3 days vs 5.5 +/- 0.5 days). The results noted with PPS were similar to those observed in cyclosporine (10 mg/kg; n = 6)-treated animals (8.25 +/- 0.25 days). Treatment with heparin (300 U/kg/day) did not significantly prolong cardiac graft survival time, suggesting that anticoagulation is not sufficient to prolong transplant survival. Analysis of tissue histology showed diminished transplant rejection as evidenced by decreased white blood cell infiltration and cellular necrosis. CONCLUSIONS: The results of this study indicate that PPS possesses the ability to prolong cardiac transplant viability in a heterotopic cardiac transplant model, independent of its anticoagulant actions. PMID- 10452865 TI - Interpreting the judgment of surgical faculty regarding resident competence. AB - BACKGROUND: It is reasonable to propose that competence is a multifaceted characteristic defined in part by some minimum level of knowledge and skill. In this study we examined the relationship between surgical faculty's judgment of clinical competence, as measured by a surgical resident objective structured clinical examination (OSCE), and the residents' objective performance on the skills being tested. METHODS: Fifty-six general surgery residents at all levels of training participated in a 30-station OSCE. At the completion of each station, the faculty proctor made several overall judgments regarding each resident's performance, including a global judgment of competent or not competent. The competence judgment was applied to the objective percentage performance score in three different ways to construct methods for determining competence based solely upon this objective percentage score. RESULTS: The average mean competent score (MCS) across the stations was 61%, and the average mean noncompetent score (MNCS) was 38%. The difference between MCS and MNCS for each station was very consistent. Upper threshold scores above which a judgment of competent was always made, and lower threshold scores below which a judgment of noncompetent was always made were observed. Overall, the average mean and threshold scores for competent and noncompetent groups were remarkably similar. For performance scores in the range between the threshold competent and noncompetent scores at each station, measures other than objective performance on the skills being evaluated determined the judgment of competent or not competent. CONCLUSIONS: Empirically determined minimum acceptable standards for objective performance in clinical skills and knowledge appeared to have been subconsciously applied to the competence judgment by the faculty evaluators in this study. Other factors appeared to have become determinate when the objective performance score fell within a range of uncertainty. PMID- 10452866 TI - Expression of HSP70 in healing wounds of diabetic and nondiabetic mice. AB - BACKGROUND: Heat shock proteins (HSPs) stabilize intracellular processes of cells under stress. Little is known about the role of HSPs in wound healing, or whether their expression is altered by systemic disease. The focus of this study was to examine the local heat shock response to wounding in diabetic mice. METHODS: Congenitally diabetic and phenotypically normal mice underwent standardized full thickness cutaneous wounding. Mice were sacrificed at sequential time points and the wound beds excised. Tissues underwent immunohistochemical (IHC) and RT-PCR analyses for inducible HSP70. RESULTS: HSP70 protein expression in the wound bed by IHC peaked at 24 h in the nondiabetic mice. Expression of HSP70 was delayed in the diabetic mice until Day 3, which correlates with the clinical delay in healing seen in this model. The protein was especially prominent in the epithelium and in inflammatory cells migrating into the granulation tissue matrix. RT-PCR demonstrated upregulation of HSP70 mRNA within 12 h after wounding, lasting until Day 3, and decreasing thereafter in both the nondiabetic and the diabetic animals. CONCLUSION: Cutaneous wounding produces a HSP response in inflammatory cells, and expression of inducible HSP70 is delayed in diabetic mice. This delay may be related to the impaired inflammatory response of diabetics, and may contribute to impaired wound healing. The wound may be a continuing source of the heat shock response in inflammatory cells after injury. PMID- 10452867 TI - Cardiopulmonary bypass renders patients at risk for multiple organ failure via early neutrophil priming and late neutrophil disability. AB - BACKGROUND: Cardiopulmonary bypass (CPB) is associated with a systemic inflammatory response syndrome (SIRS) and these patients are recognized to be at increased risk for delayed infectious complications. We have documented that circulating neutrophils (PMNs) from patients manifesting SIRS have evidence of early postinjury priming for cytotoxicity. Consequently, we hypothesized that CPB would result in early postoperative PMN hyperresponsiveness (priming). MATERIALS AND METHODS: Six patients (mean age 50 +/- 2.9 years) who underwent CPB for CABG had sequential blood samples obtained perioperatively. PMNs were isolated and superoxide anion (O(-)(2)) generation (nmol O(-)(2)/3.75 x 10(5) PMNs/min) was measured by reduction of cytochrome c after exposure to fMLP, C5a, or PMA; elastase release (% total PMN elastase content) was measured by cleavage of AAPV pNA after exposure to fMLP or C5a. RESULTS: PMNs were activated for increased elastase release 6 h after initiation of CPB. Significant PMN priming for O(-)(2) production was discovered at 3, 6, and 12 h following CPB and for elastase release at 3 and 6 h after CPB. At 2 to 3 days after CPB, O(-)(2) generation was significantly less than that of the preoperative control. Neutrophil primability with PAF was detected at 6 h after CPB. A similar defect in PAF-primable O(-)(2) production was seen 2 and 3 days post-CPB. Direct PMN interrogation with the receptor-independent activator PMA revealed loss of integrity of the NADPH oxidase at 2 and 3 days following CPB. CONCLUSIONS: A vulnerable window exists between 3 and 12 h after CPB when PMNs are primed for enhanced cytotoxicity via O(-)(2) production and elastase release. Paradoxically, PMN oxidase integrity becomes deficient 48 h post-CPB, while protease degranulation remains intact. These events render the bypass patient at risk for multiple organ failure via both early PMN-mediated tissue injury and delayed infectious complications. PMID- 10452868 TI - Products of cyclooxygenase-2 catalysis regulate postoperative bowel motility. AB - Laparotomy involving manipulation of the small intestine causes injury, initiating an inflammatory cascade in the small bowel wall, which generates eicosanoids and proinflammatory cytokines. We have shown that ketorolac and salsalate, nonselective cyclooxygenase (COX) inhibitors, ameliorate postoperative small bowel ileus in a rodent model. Others have shown that interleukin-1 receptor antagonism improves postoperative gastric emptying. We examined whether inhibition of the proinflammatory cytokines, tumor necrosis factor alpha (TNFalpha) and interleukin-1 (IL-1), or selective blockade of cyclooxygenase-2 (COX-2), the COX isoform induced during inflammation, would accelerate postoperative small bowel transit in our model. Duodenostomy tubes were inserted into male Sprague-Dawley rats. One week later, animals were randomized to receive TNF-binding protein (TNF-bp), IL-1 receptor antagonist (IL-1ra), or saline (NS) prior to standardized laparotomy. Additional rats were gavaged preoperatively with a selective COX-2 inhibitor (NS-398) or NS. Small intestinal transit was measured as the geometric center (GC) of distribution of (51)CrO(4) at 30 min, 3 h, or 6 h (n = 5-9 rats/group) following laparotomy. Selective inhibition of COX 2 significantly increased postoperative small bowel transit compared to controls (GC 2.9 +/- 0.3 vs 2.2 +/- 0.1 at 30 min, GC 2.9 +/- 0.3 vs 2.5 +/- 0.2 at 3 h, and GC 3.3 +/- 0.3 vs 2.8 +/- 0.2 at 6 h, P < 0.05). In contrast, neither TNF-bp nor IL-1ra altered postoperative small intestinal transit in this model. Use of selective COX-2 inhibitors may accelerate recovery of postoperative bowel dysmotility without the undesirable effects (e.g., gastrointestinal irritation and anti-platelet effect) of nonselective COX inhibitors. PMID- 10452870 TI - Near-infrared spectroscopic assessment of tissue hydration following surgery. AB - Near-infrared reflectance spectroscopy and imaging were used to assess tissue hydration following elevation of reverse McFarlane dorsal rat skin flaps (N = 6). A quantitative measure of tissue water content was derived using the integrated intensities of the strong absorption bands of water centered at 970 and 1450 nm. Near-infrared spectroscopy monitored tissue hydration at discrete locations, while imaging provided hydration maps of cutaneous tissues. Both methods were rapid and noninvasive. The near-infrared results were compared to laser Doppler flux measurements. Significant changes in tissue hydration were observed upon surgical elevation with substantial regional variation along the skin flap. These results indicate the potential of near-infrared spectroscopy and imaging to monitor tissue hydration changes in the skin following surgery. PMID- 10452869 TI - Change of liver function in hypertrophying lobe of rabbit liver after portal branch ligation. AB - Preoperative portal embolization (PE) is useful for the prevention of postoperative liver failure after extended hepatectomy. However, clinical evaluation of liver function in the hypertrophying lobe after PE has not been studied. Here we report functional changes in the hypertrophying lobe using a 80% portal-branch-ligation rabbit model. Liver function was evaluated by the expression of liver-specific genes detected by Northern blot analysis and plasma disappearance rate of indocyanine green (ICG). The weight of the unligated lobe after portal ligation increased about twofold on the 7th postoperative day (POD) and about threefold on the 14th POD. The mRNA levels of the liver-specific genes (albumin, aldolase B, and tyrosine aminotransferase) in the unligated lobe decreased to about 50% on the 1st POD and returned to the preoperative levels on the 7-14th POD. In contrast, the expression of histone H2B mRNA increased on the 3rd-7th POD. The plasma disappearance rate of ICG (K-ICG) in the rabbit that has only the unligated lobe did not significantly change during the first 7 days, but then improved and recovered to 80% of that in the rabbit that has whole liver on the 14th POD. These results indicate that liver function of the hypertrophying lobe after portal branch ligation does not increase during the first 7 days despite an increase in liver weight. This finding suggests that the compensatory hypertrophying liver is enlarging without functional augmentation in the early period after PE. PMID- 10452871 TI - Hepatic apoptotic activity following transient normothermic inflow occlusion and reperfusion in the swine model. AB - Accelerated hepatic apoptosis was first described in portal vein-ligated livers but has since been reported in a variety of liver injuries. Because porto-prival states can induce apoptosis we sought to determine whether transient ischemic periods followed by reperfusion would trigger such cell death. The cytokines TNF alpha and TGF-beta are known to facilitate apoptosis and are released in response to a number of stimuli including ischemia. We also investigated alterations in plasma and tissue levels of these cytokines which might lend support to their role in increased apoptotic activity following ischemia/reperfusion. Female pigs were used as the experimental model. Inflow occlusion of portal and hepatic arterial blood was performed to a portion of the swine liver directing the entire splanchnic flow to the remaining hepatic lobes for a period of 2 h. The livers were then reperfused and plasma and tissue samples taken for determination of apoptotic activity utilizing cell death immunoperoxidase staining of 3'-OH DNA ends generated by fragmentation and ELISA assay of histone-associated DNA fragments. Plasma and tissue levels of TNF-alpha and plasma levels of TGF-beta were determined by ELISA assay. An increase in apoptotic activity following reperfusion was seen at Day 2 and Day 4 compared to preischemic values by the cell death stain. The ELISA cell death assay showed an increase in apoptotic activity at 60 min, Day 2, and Day 4. Moreover, the ELISA cell death assay showed enhanced apoptotic activity in "hyperperfused" hepatic lobes compared to preischemic, or resting, liver. This was also observed when compared to sham operated animals. Surprisingly, there was no detectable increase in plasma TNF alpha or TGF-beta levels following ischemia/reperfusion, although homogenized liver TNF-alpha levels were increased at 60 min and Day 2 following reperfusion. We conclude that transient hepatic inflow occlusion followed by reperfusion can induce increased apoptotic activity in the swine model. Furthermore, increased apoptotic activity also occurs in the hyperperfused liver raising the possibility of a locally active factor or global hepatic expression of receptor activity in response to ischemia/reperfusion. This period of ischemia/reperfusion did not produce a detectable increase in circulating cytokine levels, and accelerated apoptosis could not be linked to heightened TNF-alpha or TGF-beta plasma activity. Higher tissue levels of TNF-alpha could reflect enhanced binding to TNF cell surface receptors or amplified receptor expression. PMID- 10452872 TI - Predictability of dielectric properties for ischemic injury of the skeletal muscle before reperfusion. AB - OBJECTIVE: The purpose of this study is to assess whether or not dielectric properties could predict the severity of ischemic injury of the skeletal muscle before reperfusion to prevent the myonephropathic metabolic syndrome (MNMS). METHODS: Japanese White rabbits were anesthetized and their hind limbs were subjected to 0, 2, 4, and 6 h of ischemia and then 10 h of reperfusion (n = 4, 5, 6, and 6). Dielectric properties of the gastrocnemius muscle were measured and tan delta(m) and D were calculated from them. To assess ischemic injury, we also measured (a) creatine phosphate kinase (CPK) and aldolase, (b) spectrophotometric assay of triphenyltetrazolium chloride assay, and (c) histological evaluation by H & E staining, 10 h after reperfusion. RESULTS: The tan delta(m) increased and D decreased significantly compared with those in preischemia (P < 0.01). The tan delta(m) value just before reperfusion significantly correlated positively with the maximum value of CPK (CPK(max)) or the maximum value of aldolase (ALD(max)) (r = 0.73, 0.69) and negatively with %TTC (r = 0.66). The D value just prior to reperfusion significantly correlated negatively with CPK(max) and ALD(max) (r = 0.81, 0.86) and positively with %TTC (r = 0.79) after reperfusion, respectively. Histologically, skeletal muscle necrosis was detected in 11 out of 12 skeletal muscles with tan delta(m) values which were 3.70 or more and 11 out of 14 muscles with D values which were 1.47 or less. CONCLUSION: The severity of skeletal muscle ischemic injury can be evaluated before reperfusion by measuring the dielectric properties during ischemia. PMID- 10452873 TI - Pacing-induced cardiac failure of hypertrophic hearts: effects of cyclic GMP reduction. AB - BACKGROUND: We tested the hypothesis that pacing-induced cardiac failure of hypertrophic hearts would reduce the functional and metabolic responses of these hearts to guanylate cyclase inhibition and this was associated with alterations in cyclic GMP. MATERIALS AND METHODS: Methylene blue (MB, 2 mg/kg/min, guanylate cyclase inhibitor) was infused into the left anterior descending coronary artery in 5 control, 5 left ventricular hypertrophy (LVH), and 5 LVH pacing-induced failure dogs. Regional myocardial work was calculated as the integrated product of force and segment shortening and regional myocardial O(2) consumption (VO(2)) from coronary blood flow and O(2) extraction measurements. Cyclic GMP was determined by radioimmunoassay. RESULTS: MB increased regional work (635 +/- 169 vs 1649 +/- 500, 781 +/- 184 vs 1569 +/- 203 g * mm/min) and VO(2) (8.3 +/- 1.4 vs 10.9 +/- 1.4, 7.3 +/- 0.7 vs 9.1 +/- 0.7 ml O(2)/min/100 g) in both control and LVH dogs but not in failure dogs (536 +/- 234 vs 623 +/- 193, 3.6 +/- 1.1 vs 4.7 +/- 1.9). MB also decreased cyclic GMP in control dogs (1170 +/- 142 vs 812 +/- 105 pmol/g). LVH dogs had elevated baseline cyclic GMP (5875 +/- 949) compared to control dogs but also demonstrated decreased cyclic GMP in response to MB (2820 +/- 372). In failure dogs, basal cyclic GMP was also elevated (4650 +/- 613) compared to control dogs but there was a lack of response to MB (3670 +/ 640). CONCLUSIONS: We conclude that the myocardial function, VO(2) and cyclic GMP responses to methylene blue are diminished in the transition from hypertrophy to cardiac failure. PMID- 10452874 TI - Use of cefazolin microspheres to treat localized methicillin-resistant Staphylococcus aureus infections in rats. AB - BACKGROUND: In a previous study, the topical administration of biodegradable, controlled-release poly-(dl-lactide-co-glycolide) cefazolin microspheres could effectively prevent surgical wound infections with a sensitive strain of Staphylococcus aureus in an experimental animal model. The objective of the current study was to evaluate and compare the efficacy of topical antibiotic therapy with cefazolin microspheres to systemic cefazolin therapy for the treatment of experimental rat surgical wounds contaminated with a methicillin resistant strain of S. aureus (MRSA). METHODS: A local infection model in rats was used. MRSA was used to infect pockets surgically produced in the paraspinous muscles. Groups of rats received either topical cefazolin microspheres, topical cefazolin powder, parenteral cefazolin, or no treatment. Feces were cultured to evaluate the effect of antibiotic therapy on gut flora. RESULTS: The rate of clinical wound infection following topical application of cefazolin microspheres (13%) was significantly lower than the 53% infection rate observed in rats who had received a 2-week course of systemic cefazolin therapy (P = 0.046). Moreover, single-dose topical antibiotic therapy with cefazolin microspheres completely eradicated MRSA from the wounds of 7 of 15 (47%) animals. There was no statistically significant difference, however, in the rate of clinical wound infection between rats whose wounds were treated topically with free cefazolin powder and those treated with systemic cefazolin (P = 0.12). Importantly, selection of antibiotic-resistant bacteria was associated with systemic but not local cefazolin therapy. CONCLUSION: The results of this study suggest that topical antibiotic therapy with controlled-release cefazolin microspheres may be effective for the prevention of wound infection with both methicillin-sensitive and methicillin-resistant strains of S. aureus in selected surgical procedures that are at high risk of developing postoperative wound infection. PMID- 10452875 TI - Fish oil augments macrophage cyclooxygenase II (COX-2) gene expression induced by endotoxin. AB - BACKGROUND: Fish oil-supplemented diets have anti-inflammatory and immunomodulating effects. Although fish oil is readily incorporated into the cell membrane and influences the production of eicosanoids, the exact mechanism is not clear. This study was designed to investigate the effects of eicosapentaenoic acid (EPA), a major component of fish oil, on macrophage (Mphi) cyclooxygenase (COX) gene expression induced by LPS. METHODS: RAW 264.7 cells, a mouse Mphi cell line, were grown in EPA-rich media for 24 h. Mphi were washed and exposed to Escherichia coli LPS (10 microg/ml). Membrane lipid profile was determined by gas chromatographic analysis. COX-1 and COX-2 mRNA expressions were determined by Northern blot assays with mouse-specific cDNA probes. PGE(2) production of Mphi was measured by ELISA. Mphi production of COX-2 protein was determined by Western blot assays with an anti-COX-2 antibody. RESULTS: Incubation in EPA-rich media increased membrane EPA and decreased arachidonic acid (AA) composition. COX-2 mRNA expression was induced by EPA and further augmented by LPS stimulation. EPA also augmented Mphi production of COX-2 protein. In comparison, COX-1 mRNA expression was not affected by either LPS stimulation or EPA incubation. EPA reduced PGE(2) production by LPS-stimulated Mphi. To further support that COX-2 mRNA was regulated by COX product, exogenous PGE(2) was added to Mphi prior to LPS stimulation. PGE(2) reduced COX-2 mRNA of LPS-stimulated Mphi. CONCLUSION: EPA displaces AA and reduces PGE(2) production by LPS-stimulated Mphi. Fish oil inhibition of Mphi PGE(2) production induces COX-2 mRNA expression through a COX 2 product-mediated feedback mechanism. PMID- 10452876 TI - Expression of gastrin/CCK-B receptors does not lead to a mitogenic response to gastrin in two colon cell lines. AB - To clarify the impact of the classic gastrin/CCK-B receptor on the growth of benign and malignant colonic cells, two permanent cell lines expressing this receptor have been established. The conditionally immortalized nonmalignant colonic cell line YAMC and the colonic carcinoma cell line SW 403 were stably transfected with a plasmid encoding the gastrin/CCK-B receptor (GR), or with plasmid alone (V). Expression of the gastrin/CCK-B receptor in the transfected YAMC-GR and SW 403-GR cells was demonstrated by gastrin binding experiments. The YAMC-GR cell line did not respond mitogenically to pentagastrin or gastrin(17) in vitro and was not tumorigenic. The SW 403-GR cell line was stimulated by gastrin(17) in vitro, but the growth patterns of SW 403-GR and SW 403-V were the same in nude mice with cells injected either subcutaneously or into the spleen. These results provide further evidence that the gastrin/CCK-B receptor is not responsible for gastrin-stimulated growth in colonic tumors. PMID- 10452877 TI - Acute portal hypertension increases ileal vulnerability to platelet-activating factor in rats. AB - BACKGROUND: Patients with portal hypertension can easily develop sepsis of enteric origin after suffering severe trauma and hemorrhagic shock. Platelet activating factor (PAF) is one of the key mediators of such stress. The aim of this study was to investigate whether portal hypertension increases the vulnerability of the ileum to PAF. MATERIALS AND METHODS: Seven days after surgery, PAF (1.5 microg/kg) was intravenously injected into portal stenosis (PS) rats and sham-operated rats. The levels of tumor necrosis factor-alpha (TNF alpha), cytokine-induced neutrophil chemoattractant (CINC), and endotoxin in portal plasma were determined. The levels of PAF receptor (PAFR), TNF-alpha, and CINC mRNA in the ileum were also investigated. RESULTS: After PAF administration, PS rats showed (1) significantly higher portal plasma levels of TNF-alpha, CINC, and endotoxin; (2) higher histological damage scores in the ileum; (3) more infiltrating neutrophils in the ileum; and (4) a significantly higher mortality rate than sham-operated rats (P < 0.01). However, PAFR mRNA levels were similar in the two groups. The CINC mRNA level in the ileum of PS rats was increased from 1 to 4 h after PAF administration, while that of the sham-operated rats was transiently increased at 1 h. CONCLUSIONS: Portal hypertension increases the vulnerability of the ileum to PAF. These findings suggest that conditions which causes PAF production may be dangerous in patients with portal hypertension. PMID- 10452878 TI - Pressure volume curves in arrested heterotopic rat heart isografts: role of improved myocardial protection. AB - BACKGROUND: To minimize decreases in left ventricular (LV) compliance immediately after rat heart transplantation, we tested several different methods of myocardial protection. MATERIALS AND METHODS: Five groups of ACI rat hearts (n = 6 each) were arrested by coronary perfusion with 5 ml of UW (University of Wisconsin), UW-BDM (UW with 2,3-butanedione monoxime), CU (Columbia University), or CU-BDM solution or by LV injection of potassium chloride and Ringer's lactate immersion (KCl/RL). After abdominal isografting and blood reperfusion for 15 min, transplanted hearts (TxH) were arrested and excised. Diastolic LV pressure-volume curves (LVPVCs) were correlated with myocardial water content (MWC). Native hearts (NH) were arrested identically to TxH and maintained at 4 degrees C by immersion. LVPVCs were measured at 15-min intervals for 90 min. RESULTS: In three of four pressure intervals at Time 0, normalized LV volume (LVV) was smaller (P < 0.05, ANOVA) in KCl/RL native hearts than in the four perfusion groups. LVV decreased significantly in NH after 45-75 min; LVV decreased similarly with time in all groups. In TxH, postarrest LVVs were higher with UW-BDM, CU-BDM, and CU than with UW or KCl/RL (P < 0.05, ANOVA). Expressing LVV of TxH as a percentage of NH, UW-BDM, CU, and CU-BDM provided qualitatively better diastolic properties than KCl/RL and UW. CONCLUSIONS: Thus rat LVPVCs can be improved after heart transplantation with alternative strategies of myocardial protection. KCl arrest decreases LV filling volume in this model and should be avoided. PMID- 10452879 TI - 13CO(2) peak value of L-[1-(13)C]phenylalanine breath test reflects hepatopathy. AB - BACKGROUND: Using a rat model of hepatectomy, we investigated whether the severity of hepatopathy could be quantitatively measured from changes in expiratory (13)CO(2) levels after intravenous administration of L-[1 (13)C]methionine or L-[1-(13)C]phenylalanine. MATERIALS AND METHODS: Under nembutal anesthesia, 30 mg/kg L-[1-(13)C]phenylalanine or 40 mg/kg L-[1 (13)C]methionine was administered to rats through the femoral vein, and expiratory (13)CO(2) levels were measured for 15 min. A 30, 70, or 90% hepatectomy was performed. In the control group, simple laparotomy was performed. Breath test was conducted 20 min after laparotomy. We examined the correlation of the total (13)CO(2) output over 15 min or peak (13)CO(2) level with liver weight/body weight (%). RESULTS: In breath test graphs, L-[1-(13)C]methionine did not show any peak level during measurement. L-[1-(13)C]phenylalanine showed a specific peak level 6 +/- 1 min after administration. The correlation coefficient between total (13)CO(2) output over 15 min after L-[1-(13)C]methionine administration and liver weight/body weight was 0.922 (P < 0.001). The correlation coefficient between total (13)CO(2) output over 15 min after L-[1 (13)C]phenylalanine administration and liver weight/body weight was 0.883 (P < 0.001). The correlation coefficient between peak L-[1-(13)C]phenylalanine level and liver weight/body weight was highest, 0.927 (P < 0.001). CONCLUSION: In a breath test with intravenously administered L-[1-(13)C]methionine or L-[1 (13)C]phenylalanine, hepatopathy could be quantitatively evaluated by measuring expiratory (13)CO(2) levels over 15 min. After administration of L-[1 (13)C]phenylalanine, hepatopathy could be quantitatively evaluated in a short period by measuring the peak expiratory (13)CO(2) level. PMID- 10452880 TI - Early detection of apoptosis and fas ligand expression in hepatic graft-versus host disease after rat small bowel transplantation. AB - BACKGROUND: The liver is one of the primary targets of acute graft-versus-host disease (GVHD), which is the principal complication that occurs after allogeneic intestinal transplantation. The purpose of this study is to investigate the involvement of the Fas/Fas ligand system in hepatic GVHD after rat semiallogeneic intestinal transplantation. MATERIALS AND METHODS: Liver samples were serially harvested from LEW x BN F(1) (LBNF(1)) recipients of either LEW heterotopic intestinal allografts (group 1) or LBNF(1) isografts (group 2), on Days 1, 3, 5, 9, and 13 posttransplant. RESULTS: In group 1, hepatic injuries as assessed by either serum aspartate aminotransferase (AST) level, alanine aminotransferase (ALT) level, or cellular infiltration on HE staining became apparent after Day 13. The incidence of apoptosis, examined by terminal deoxynucleotidyl transferase mediated dUTP biotin nick end-labeling (TUNEL), was observed to steadily increase in the liver from Day 5 accompanied by a progression of GVHD: 17.5 +/- 3.1 and 3.1 +/- 0.4 cells/field (200x) in groups 1 and 2, respectively. In an immunohistochemical study, Fas was constitutively expressed in the liver in both groups, while Fas ligand was expressed most extensively on Day 13 in group 1. Immunoreactivity of both Fas and Fas ligand was observed in hepatocytes, in addition to leukocytes. Analysis by reverse transcription polymerase chain reaction also revealed the expression of Fas mRNA to be constitutive in both groups, while that of Fas ligand mRNA increased significantly from Day 5 and peaked on Day 13 in group 1, and the expression was 10 times stronger than that for isogeneic combination (group 2). CONCLUSION: Early detection of upregulated Fas ligand and increased apoptosis is thus considered to be potentially a useful tool for the diagnosis of hepatic GVHD. PMID- 10452881 TI - Addition of a water-soluble alpha-tocopherol analogue to University of Wisconsin solution improves endothelial viability and decreases lung reperfusion injury. AB - BACKGROUND: Reperfusion injury following lung preservation has been associated with free radical formation and subsequent endothelial cell damage. Trolox is a water-soluble analogue of the free radical scavenger alpha-tocopherol. We hypothesized that addition of this form of vitamin E to University of Wisconsin (UW) solution would decrease reperfusion injury and improve lung function after cold ischemic preservation. MATERIALS AND METHODS: Bovine aortic endothelial cells were cultured and stored at 4 degrees C for 12, 24, and 48 h in UW or UW + Trolox (UWT). Endothelial cell viability after storage was assessed by dimethylthiazole tetrazolium cytotoxicity assay. An isolated rat perfused lung (IPL) model was used and lungs were flushed with the respective solutions with cold storage times of 6 and 12 h. Following storage, the lungs were reperfused with fresh blood and lung function was assessed by blood gas analysis, alveolar arterial gradient, and compliance. RESULTS: There was no difference in endothelial cell viability between UW and UWT after 12 or 24 h; however, UWT had higher endothelial cell viability than UW with 48 h of cold ischemic storage. Using the IPL model, the pO2 was higher with UWT than UW after 6 and 12 h of cold ischemia. The alveolar-arterial oxygen difference was significantly lower for UWT versus UW at 6 h. UWT provided increased compliance at 6 and 12 h of ischemia. CONCLUSIONS: The addition of a water-soluble vitamin E analogue to UW solution resulted in increased endothelial cell viability after prolonged storage and improved whole lung preservation in the postreperfusion period as evidenced by higher oxygenation and increased compliance. These results are clinically relevant as the lung is extremely sensitive to reperfusion injury and UW solution is being increasingly used in lung transplantation and remains the predominant solution in abdominal organ transplantation. PMID- 10452883 TI - The role of antioxidant enzymes in the control of opossum gallbladder motility. AB - BACKGROUND: Superoxide rapidly oxidizes nitric oxide (NO) to form peroxynitrite, thus terminating the biological activity of NO. The aims of our study were to determine if superoxide alters the motor function of the gallbladder and to localize the antioxidant enzymes in the gallbladder. MATERIALS AND METHODS: Immunostaining and immunoblots were performed and enzyme activities were measured in the gallbladder. In physiologic experiments, force-displacement transducers recorded tension in gallbladder muscle strips. Superoxide was generated by the addition of xanthine with xanthine oxidase, while superoxide radicals were scavenged by the addition of superoxide dismutase (SOD) and catalase. SOD was inhibited by deithyldithiocarbamate. RESULTS: Immunostaining demonstrated superoxide dismutase and catalase immunoreactivity in ganglia situated throughout the smooth muscle. Total superoxide dismutase activity was 115 +/- 12 U/mg. Western blots detected expression of proteins of 19.4 kDa for copper/zinc SOD and 25.0 kDa for manganese SOD. Generation of superoxide increased isometric tension, while pretreatment with SOD prevented the increase in isometric tension induced by superoxide. Inhibition of SOD diminished the EFS-induced off response. CONCLUSIONS: We conclude that superoxide alters gallbladder motor function, and the presence of superoxide scavenging enzymes in enteric plexuses suggests that they may regulate gallbladder neuromuscular function by clearing endogenous superoxide. PMID- 10452882 TI - Effect of specific neutrophil elastase inhibitor on ischemia/reperfusion injury in rat liver transplantation. AB - Activated neutrophils have been implicated as playing an important role in ischemia/reperfusion injury of the liver by releasing toxic mediators such as oxygen free radicals and elastases. In the present study, we evaluated the effect of a novel, specific neutrophil elastase inhibitor (ONO-5046) on cold ischemia/reperfusion injury of the liver allograft in rodents. Livers from male Lewis rats were procured and stored cold (4 degrees C) in lactated Ringer's solution and transplanted orthotopically. Recipients were divided into three groups: Vehicle group, 5-h preservation and vehicle (n = 8); ONO-5046 group, 5-h preservation and administration of ONO-5046 (n = 8); and Control group, minimum preservation only (n = 8). Bile output after reperfusion was significantly larger in the ONO-5046 group compared to the Vehicle group (P < 0.05 or less). Sinusoidal endothelial cell function represented by the serum hyaluronic acid concentration at 120 min after reperfusion of the ONO-5046 group was significantly lower than that in the Vehicle group (17.0 +/- 7.9 vs 36.2 +/- 14.9 ng/ml, P < 0.05), whereas serum transaminase levels 120 min after reperfusion were comparable between the two groups. Liver tissue energy charge 120 min after reperfusion was significantly better in the ONO-5046 group compared to the Vehicle group (P < 0.05). Furthermore, the number of neutrophils infiltrating the allograft after reperfusion was significantly depressed in the ONO-5046 group compared to the Vehicle group (P < 0. 02). These data suggest that the neutrophil elastase might cause liver damage early after reperfusion in cold-stored liver, which can be ameliorated by the administration of a specific neutrophil elastase inhibitor, ONO-5046. PMID- 10452884 TI - Which technique, circular stapled anastomosis or double stapling anastomosis, provides the optimal size and shape of rectal anastomotic opening? AB - BACKGROUND: Two types of end-to-end colorectal anastomosis using a circular stapler, circular stapled anastomosis(CSA) and double stapling anastomosis(DSA), have become the standard technique. The possibility of anastomotic openings of the DSA being smaller and somewhat distorted, because of some portion of the stapled rectal stump not being included in the anastomotic opening, was examined. METHODS: Anastomotic openings created by CSA and DSA were photographed through a sigmoidoscope, and the maximal and minimal diameters and the areas of the anastomotic openings were measured and calculated with an image analyzer. They were examined in the swine rectum immediately after surgery and were also examined in the randomized clinical cases 3 or more months after surgery. RESULTS: It was found that larger anastomotic openings could be created as the diameter of the anastomosed intestine and the cartridge size became wider in both methods immediately after surgery. The shape of anastomotic opening created by DSA was observed to be more oval than round immediately after surgery. The anastomotic stenosis occurred in a small number of patients in both methods within 3 months after surgery. The area of the anastomotic opening created by CSA was about 25% larger than that by DSA immediately after surgery and about 30% larger 3 or more months after surgery. CONCLUSION: CSA provides a larger anastomotic opening than DSA. Anastomotic stenosis occurred in a small number of patients in both methods. PMID- 10452885 TI - The tax protein-DNA interaction is essential for HTLV-I transactivation in vitro. AB - The human T-cell leukemia virus type-1 (HTLV-I)-encoded Tax protein enhances viral gene transcription through interaction with three repeated DNA elements located in the viral promoter. These elements, called viral CREs, are composed of an off-consensus eight base-pair cyclic AMP response element (CRE), immediately flanked by sequences that are rich in guanine and cytosine residues. Recent biochemical experiments have demonstrated that in the presence of the cellular protein CREB, Tax directly binds the viral CRE G+C-rich sequences via interaction with the minor groove. To determine the functional significance of the Tax-DNA interaction, we synthesized minor groove-binding pyrrole-imidazole polyamides which bind specifically to the G+C-rich sequences in the viral CREs. At concentrations where the polyamides specifically protect the G+C-rich sequences from MPE:Fe cleavage, the polyamides block the Tax-DNA interaction. At precisely these same concentrations, the polyamides specifically inhibit Tax transactivation in vitro, without altering CREB-activated transcription or basal transcription from the same promoter. Together, these data provide strong evidence that Tax-viral CRE interaction is essential for Tax function in vitro, and suggest that targeted disruption of the Tax-DNA minor groove interaction with polyamides may provide a novel approach for inhibiting viral replication in vivo. PMID- 10452886 TI - Shutdown in protein synthesis due to the expression of mini-genes in bacteria. AB - Mutants of Escherichia coli partially deficient in peptidyl-tRNA hydrolase are killed by the expression of certain very short open reading frames (mini-genes), encoded by the wild-type bar regions of phage lambda. According to the current hypothesis, protein synthesis is shut off, and the host cells die, after essential tRNA species become sequestered due to abnormal translation termination (drop-off) of mini-gene-encoded peptides as peptidyl-tRNA. Here we study variants of bar mini-genes, both in vivo and in vitro, in order to identify the structural elements that influence this inhibition of protein synthesis. Three parameters were measured during the expression of these variants: the rates of normal translation termination, peptidyl-tRNA dissociation from the ribosome and hydrolysis of peptidyl-tRNA by peptidyl-tRNA hydrolase were measured. Previous observations that RRF, EF-G and RF3 stimulated drop-off were confirmed and extended; stimulation by these factors can reach 30-fold. Both factor-stimulated and spontaneous drop-off depended on the nature of the stop signal. The degree of inhibition of cell growth following induction of mini-gene expression could be accounted for in terms of a toxicity index comprising the three parameters above. Inhibition was greatly reduced in cells lacking RF3. Mini-genes with more efficient Shine/Dalgarno sequences killed cells even with normal peptidyl-tRNA hydrolase activity. It is proposed that the retranslation by ribosomes of mini gene transcripts with efficient ribosome binding (Shine/Dalgarno) sequences strongly contributes to the inhibitory effects of mini-gene expression on protein synthesis. PMID- 10452888 TI - Developmental regulation of replication fork pausing in Xenopus laevis ribosomal RNA genes. AB - In early Xenopus embryos, replication forks move along the rRNA genes (rDNA) at a uniform rate and terminate at multiple, apparently random sites. In contrast, a polar replication fork barrier (RFB) is found at the 3' end of the rRNA genes in Xenopus cultured cells. We have now analysed the replication intermediates of Xenopus rDNA from a wide range of developmental stages by 2D gel electrophoresis. Surprisingly, up to 15 different replication fork pausing sites (RFPs) simultaneously appear in the rDNA at the midgastrula stage, when rRNA transcription abruptly increases. They disappear during the neurula stage, except for a polar RFP at the 3' end of Xthe transcription unit, which persists to the tadpole stage. The latter RFP is found at the same location as the RFB in cultured cells; however the arrest of replication forks at this RFP is not absolute, since termination occurs at multiple positions throughout the rDNA repeat. The efficiency of fork arrest at this RFP remains constant from midgastrula to early tadpole, and decreases around hatching. The transient appearance of multiple RFPs at midgastrula may reflect some chromatin remodeling associated with developmental activation of rRNA transcription. PMID- 10452887 TI - Yeast aspartyl-tRNA synthetase residues interacting with tRNA(Asp) identity bases connectively contribute to tRNA(Asp) binding in the ground and transition-state complex and discriminate against non-cognate tRNAs. AB - Crystallographic studies of the aspartyl-tRNA synthetase-tRNA(Asp)complex from yeast identified on the enzyme a number of residues potentially able to interact with tRNA(Asp). Alanine replacement of these residues (thought to disrupt the interactions) was used in the present study to evaluate their importance in tRNA(Asp)recognition and acylation. The results showed that contacts with the acceptor A of tRNA(Asp)by amino acid residues interacting through their side chain occur only in the acylation transition state, whereas those located near the G73 discriminator base occur also during initial binding of tRNA(Asp). Interactions with the anticodon bases provide the largest free energy contribution to stability of the enzyme-tRNA complex in its ground state. These contacts also favour catalysis, by acting connectively with each other and with those of G73, as shown by multiple mutant analysis. This implies structural communication transmitting the anticodon recognition signal to the distally located acylation site. This signal might be conveyed via tRNA(Asp)as suggested by the observed conformational change of this molecule upon interaction with AspRS. From binding free energy values corresponding to the different AspRS tRNA(Asp)interaction domains, it might be concluded that upon complex formation, the anticodon interacts first. Finally, acylation efficiencies of AspRS mutants in the presence of pure tRNA(Asp)and non-fractionated tRNAs indicate that residues involved in the binding of identity bases also discriminate against non cognate tRNAs. PMID- 10452889 TI - Gene organisation, sequence variation and isochore structure at the centromeric boundary of the human MHC. AB - We have mapped and sequenced the region immediately centromeric of the human major histocompatibility complex (MHC). A cluster of 13 genes/pseudogenes was identified in a 175 kb PAC linking the TAPASIN locus with the class II region. It includes two novel human genes (BING4 and SACM2L) and a thus far unnoticed human leucocyte antigen (HLA) class II pseudogene, termed HLA-DPA3. Analysis of the G+C content revealed an isochore boundary which, together with the previously reported telomeric boundary, defines the MHC class II region as one of the first completely sequenced isochores in the human genome. Comparison of the sequence with limited sequence from other cell lines shows that the high sequence variation found within the classical class II region extends beyond the identified isochore boundary leading us to propose the concept of an "extended MHC". By comparative analysis, we have precisely identified the mouse/human synteny breakpoint at the centromeric end of the extended MHC class II region between the genes HSET and PHF1. PMID- 10452890 TI - Domain organization of Mac-2 binding protein and its oligomerization to linear and ring-like structures. AB - The multidomain Mac-2 binding protein (M2BP) is present in serum and in the extracellular matrix in the form of linear and ring-shaped oligomers, which interact with galectin-3, fibronectin, collagens, integrins and other large glycoproteins. Domain 1 of M2BP (M2BP-1) shows homology with the cysteine-rich SRCR domain of scavanger receptor. Domains 2 and 3 are related to the dimerization domains BTB/POZ and IVR of the Drosophila kelch protein. Recombinant M2BP, its N-terminal domain M2BP-1 and a fragment consisting of putative domains 2, 3 and 4 (M2BP-2,3,4) were investigated by scanning transmission electron microscopy, transmission electron microscopy, analytical ultracentrifugation and binding assays. The ring oligomers formed by the intact protein are comprised of approximately 14 nm long segments composed of two 92 kDa M2BP monomers. Although the rings vary in size, decamers predominate. The various linear oligomers also observed are probably ring precursors, dimers predominate. M2BP-1 exhibits a native fold, does not oligomerize and is inactive in cell attachment. M2BP-2,3,4 aggregates to heterogeneous, protein filled ring-like structures as shown by metal shadowed preparations. These aggregates retain the cell-adhesive potential indicating native folding. It is hypothesized that the rings provide an interaction pattern for multivalent interactions of M2BP with target molecules or complexes of ligands. PMID- 10452891 TI - Nucleosome dynamics. Protein and DNA contributions in the chiral transition of the tetrasome, the histone (H3-H4)2 tetramer-DNA particle. AB - Our laboratory has previously reported the chiral transition of DNA minicircle reconstituted tetrasomes (the particles made of DNA wrapped around the histone (H3-H4)2tetramer). This transition was induced by DNA positive torsional constraint, generated either by initial supercoiling of the loop or by its thermal fluctuations during topoisomerase relaxation. Taking into account the wrapping of the DNA around the histones into less than a turn, and its negative crossing at the entry-exit, the transition was proposed to involve a 360 degrees rotation of the loop around the particle dyad axis, and the formation of a positive crossing. The tetramer horseshoe-shaped conformation within the octamer further suggested that this process could be mediated by a reorientation of the two sector-like H3-H4 dimers about their H3/H3 interface, which would switch the overall handedness of the proteinaceous superhelix from left to right-handed. We now provide additional evidence for such a contribution of the protein by showing, through gel electrophoresis, topoisomerase relaxation and electron microscopy, that a sterical hindrance at the H3/H3 interface, introduced by covalent linking of bulky adducts through thiol oxidation of H3 cysteine 110, interferes with the transition. Such interference varies, depending on the particular SH-reagent used; but the most remarkable effect was obtained with 5, 5'-dithiobis (2-nitrobenzoic acid) (DTNB), which displaces the preferred conformation of the tetrasomes from left-handed to semi-right-handed, and at the same time preserves a significant degree of chiral flexibility. DNA contribution was evidenced by a specific fractionation of circular tetrasomes in gel electrophoresis which, together with a different positioning of control and DTNB tetrasomes on linear DNA, pointed to an interdependence between tetrasome conformation and positions. Moreover, linear tetrasomes fluctuate between crossed and uncrossed conformations in a salt-dependent equilibrium which appears to vary with their positions on the DNA. These data suggest a modulatable role of the DNA around the dyad in the transition, depending primarily on its sequence-dependent deformability. This role is played at both levels of H3-H4 dimer reorientation and lateral opening, a mechanism by which the particle may relieve the clash between its entering and exiting DNAs. These properties make the tetrasome an attractive potential intermediate in nucleosome dynamics in vivo, in particular duringX transcriptional activation and elongation. PMID- 10452892 TI - Mutational analysis of the thermostable arginine repressor from Bacillus stearothermophilus: dissecting residues involved in DNA binding properties. AB - Recently the crystal structure of the DNA-unbound form of the full-length hexameric Bacillus stearothermophilus arginine repressor (ArgR) has been resolved, providing a possible explanation for the mechanism of arginine-mediated repressor-operator DNA recognition. In this study we tested some of these functional predictions by performing site-directed mutagenesis of distinct amino acid residues located in two regions, the N-terminal DNA-binding domain and the C terminal oligomerization domain of ArgR. A total of 15 mutants were probed for their capacity to repress the expression of the reporter argC - lacZ gene fusion in Escherichia coli cells. Substitutions of highly conserved amino acid residues in the alpha2 and alpha3 helices, located in the winged helix-turn-helix DNA binding motif, reduced repression. Loss of DNA-binding capacity was confirmed in vitro for the Ser42Pro mutant which showed the most pronounced effect in vivo. In E. coli, the wild-type B. stearothermophilus ArgR molecule behaves as a super repressor, since recombinant E. coli host cells bearing B. stearothermophilusargR on a multicopy vector did not grow in selective minimal medium devoid of arginine and grew, albeit weakly, when l -arginine was supplied. All mutants affected in the DNA-binding domain lost this super-repressor behaviour. Replacements of conserved leucine residues at positions 87 and/or 94 in the C-terminal domain by other hydrophobic amino acid residues proved neutral or caused either derepression or stronger super-repression. Substitution of Leu87 by phenylalanine was found to increase the DNA-binding affinity and the protein solubility in the context of a double Leu87Phe/Leu94Val mutant. Structural modifications occasioned by the various amino acid substitutions were confirmed by circular dichroism analysis and structure modelling. PMID- 10452893 TI - Crystal structure of diaminopelargonic acid synthase: evolutionary relationships between pyridoxal-5'-phosphate-dependent enzymes. AB - The three-dimensional structure of diaminopelargonic acid synthase, a vitamin B6 dependent enzyme in the pathway of the biosynthesis of biotin, has been determined to 1.8 A resolution by X-ray crystallography. The structure was solved by multi-wavelength anomalous diffraction techniques using a crystal derivatized with mercury ions. The protein model has been refined to a crystallographic R value of 17.5% (R -free 22.6%). Each enzyme subunit consists of two domains, a large domain (residues 50-329) containing a seven-stranded predominantly parallel beta-sheet, surrounded by alpha-helices, and a small domain comprising residues 1 49 and 330-429. Two subunits, related by a non-crystallographic dyad in the crystals, form the homodimeric molecule, which contains two equal active sites. Pyridoxal-5'-phosphate is bound in a cleft formed by both domains of one subunit and the large domain of the second subunit. The cofactor is anchored to the enzyme by a covalent linkage to the side-chain of the invariant residue Lys274. The phosphate group interacts with main-chain nitrogen atoms and the side-chain of Ser113, located at the N terminus of an alpha-helix. The pyridine nitrogen forms a hydrogen bond to the side-chain of the invariant residue Asp245. Electron density corresponding to a metal ion, most likely Na(+), was found in a tight turn at the surface of the enzyme. Structure analysis reveals that diaminopelargonic acid synthase belongs to the family of vitamin B6-dependent aminotransferases with the same fold as originally observed in aspartate aminotransferase. A multiple structure alignment of enzymes in this family indicated that they form at least six different subclasses. Striking differences in the fold of the N-terminal part of the polypeptide chain are one of the hallmarks of these subclasses. Diaminopelargonic acid synthase is a member of the aminotransferase subclass III. From the structure of the non-productive complex of the holoenzyme with the substrate 7-keto-8-aminopelargonic acid the location of the active site and residues involved in substrate binding have been identified. PMID- 10452894 TI - High resolution crystal structures of the Escherichia coli lytic transglycosylase Slt70 and its complex with a peptidoglycan fragment. AB - The 70 kDa soluble lytic transglycosylase (Slt70) from Escherichia coli is an exo muramidase, that catalyses the cleavage of the glycosidic bonds between N acetylmuramic acid and N -acetylglucosamine residues in peptidoglycan, the main structural component of the bacterial cell wall. This cleavage is accompanied by the formation of a 1,6-anhydro bond between the C1 and O6 atoms in the N acetylmuramic acid residue (anhMurNAc). Crystallographic studies at medium resolution revealed that Slt70 is a multi-domain protein consisting of a large ring-shaped alpha-superhelix with on top a catalytic domain, which resembles the fold of goose-type lysozyme. Here we report the crystal structures of native Slt70 and of its complex with a 1,6-anhydromuropeptide solved at nominal resolutions of 1.65 A and 1.90 A, respectively. The high resolution native structure reveals the details on the hydrogen bonds, electrostatic and hydrophobic interactions that stabilise the catalytic domain and the alpha superhelix. The building-block of the alpha-superhelix is an "up-down-up-down" four-alpha-helix bundle involving both parallel and antiparallel helix pairs. Stabilisation of the fold is provided through an extensive packing of apolar atoms, mostly from leucine and alanine residues. It lacks, however, an internal consensus sequence that characterises other super-secondary helical folds like the beta-helix in pectate lyase or the (beta-alpha)-helix in the ribonuclease inhibitor. The 1, 6-anhydromuropeptide product binds in a shallow groove adjacent to the peptidoglycan-binding groove of the catalytic domain. The groove is formed by conserved residues at the interface of the catalytic domain and the alpha superhelix. The structure of the Slt70-1, 6-anhydromuropeptide complex confirms the presence of a specific binding-site for the peptide moieties of the peptidoglycan and it substantiates the notion that Slt70 starts the cleavage reaction at the anhMurNAc end of the peptidoglycan. PMID- 10452895 TI - Structure of bacteriorhodopsin at 1.55 A resolution. AB - Th?e atomic structure of the light-driven ion pump bacteriorhodopsin and the surrounding lipid matrix was determined by X-ray diffraction of crystals grown in cubic lipid phase. In the extracellular region, an extensive three-dimensional hydrogen-bonded network of protein residues and seven water molecules leads from the buried retinal Schiff base and the proton acceptor Asp85 to the membrane surface. Near Lys216 where the retinal binds, transmembrane helix G contains a pi bulge that causes a non-proline? kink. The bulge is stabilized by hydrogen bonding of the main-chain carbonyl groups of Ala215 and Lys216 with two buried water molecules located between the Schiff base and the proton donor Asp96 in the cytoplasmic region. The results indicate extensive involvement of bound water molecules in both the structure and the function of this seven-helical membrane protein. A bilayer of 18 tightly bound lipid chains forms an annulus around the protein in the crystal. Contacts between the trimers in the membrane plane are mediated almost exclusively by lipids. PMID- 10452896 TI - The ligandin (non-substrate) binding site of human Pi class glutathione transferase is located in the electrophile binding site (H-site). AB - Glutathione S -transferases (GSTs) play a pivotal role in the detoxification of foreign chemicals and toxic metabolites. They were originally termed ligandins because of their ability to bind large molecules (molecular masses >400 Da), possibly for storage and transport roles. The location of the ligandin site in mammalian GSTs is still uncertain despite numerous studies in recent years. Here we show by X-ray crystallography that the ligandin binding site in human pi class GST P1-1 occupies part of one of the substrate binding sites. This work has been extended to the determination of a number of enzyme complex crystal structures which show that very large ligands are readily accommodated into this substrate binding site and in all, but one case, causes no significant movement of protein side-chains. Some of these molecules make use of a hitherto undescribed binding site located in a surface pocket of the enzyme. This site is conserved in most, but not all, classes of GSTs suggesting it may play an important functional role. PMID- 10452897 TI - Solution structure of human GAIP (Galpha interacting protein): a regulator of G protein signaling. AB - The solution structure of the human protein GAIP (Galpha interacting protein), a regulator of G protein signaling, has been determined by NMR techniques. Dipolar couplings of the oriented protein in two different liquid crystal media have been used in the structure calculation. The solution structure of GAIP is compared to the crystal structure of an homologous protein from rat (RGS4) complexed to the alpha-subunit of a G protein. Some of RGS4 residues involved in the Galpha-RGS binding interface have similar orientations in GAIP (free form), indicating that upon binding these residues do not suffer conformational rearrangements, and therefore, their role does not seem to be restricted to Galpha interaction but also to RGS folding and stability. We suggest that other structural differences between the two proteins may be related to the process of binding as well as to a distinct efficiency in their respective GTPase activating function. PMID- 10452898 TI - Ligand binding induces a large conformational change in O-acetylserine sulfhydrylase from Salmonella typhimurium. AB - Covalent binding of L-methionine as an external aldimine to the pyridoxal 5' phosphate-cofactor in the K41A mutant of O-acetylserine sulfhydrylase from Salmonella typhimurium induces a large conformational change in the protein. Methionine mimics the action of the substrate O-acetyl-L-serine during catalysis. The alpha-carboxylate moiety of L-methionine in external aldimine linkage with the active site pyridoxal 5'-phosphate forms a hydrogen bonding network to the "asparagine-loop" P67-T68-N69-G70 which adopts a different conformation than in the native protein. The side-chain nitrogen of Asn69 moves more than 7 A to make a hydrogen bond to the alpha-carboxylate group of the inhibitor. As the external aldimine is formed, the PLP tilts by 13 degrees along its longitudinal axis such that C4' moves toward the entrance to the active site and the side-chain of the methionine is directed toward the active site entrance. The local rearrangement acts as a trigger to induce a large global conformational change in the protein. A subdomain comprised of beta-strand 4, alpha-helix 3, beta-strand 5 and alpha helix 4 moves towards the active site by a rotation of 7 degrees. This subdomain movement results in a reduction of the severe twist of its central beta-sheet and reduces the active site entrance to a small hole, giving access only to small molecules like sulfide, the second substrate, or acetate, the first product. PMID- 10452899 TI - Pressure/temperature effects on protein flexibilty from acrylamide quenching of protein phosphorescence. AB - Pressure is an effective modulator of protein structure and biological function. The influence of hydrostatic pressure (/=0.878, and only nine had correlation coefficients r>beta2/Pup1>/=beta1/Pre3. PMID- 10452903 TI - Maternal cocaine use and cigarette smoking in pregnancy in relation to amino acid transport and fetal growth. AB - This review covers the weight of evidence that shows the association of cocaine and cigarette smoking in pregnancy with the impaired transplacental amino acid transport which might give rise to fetal growth restriction (IUGR). Vasoconstrictive effects of both cocaine and nicotine on the placental vasculature are clearly not the only cause for inhibition of placental amino acid uptake and transfer. In vitro studies strongly suggest that cocaine decreases the activity of placental amino acid transport system A and system N, and possibly system l and system y(+), while nicotine decreases the activity of system A. These findings are supported by cordocentesis studies in human IUGR pregnancies not resulting from drug abuse. More work is needed to be done in order to understand the potential additive or synergistic effect of cocaine and cigarette smoking on fetal growth and to determine the underlying cellular mechanisms of interaction with placental amino acid transporters. PMID- 10452904 TI - J. P. Hill on placentation in primates. PMID- 10452905 TI - Folic acid and homocyst(e)ine metabolic defects and the risk of placental abruption, pre-eclampsia and spontaneous pregnancy loss: A systematic review. AB - Placental infarction or abruption, recurrent pregnancy loss and pre-eclampsia are thought to arise due to defects within the placental vascular bed. Deficiencies of vitamin B12 and folate, or other abnormalities within the methionine homocyst(e)ine pathway have been implicated in the development of such placental diseases. We conducted a systematic literature review to quantify the risk of placental disease in the presence of these metabolic defects. Studies were identified through OVID Medline between 1966 and February 1999. Terms relating to the measurement of vitamin B12, folic acid, methylenetetrahydrofolate reductase or homocyst(e)ine were combined with those of pre-eclampsia, placental abruption/infarction or spontaneous and habitual abortion. Human studies comprising both cases and controls and published in the English language were accepted. Their references were explored for other publications. Data were abstracted on the matching of cases with controls, the mean levels of folate, B12 or homocyst(e)ine in each group or the frequency of the homozygous state for the thermolabile variant of methylenetetrahydrofolate reductase. The definition of 'abnormal' for each exposure was noted and the presence or absence of the exposure of interest for each outcome was calculated as an absolute rate with a 95 per cent confidence interval. The crude odds ratios were calculated for each study and then pooled using a random effects model. Eighteen studies were finally included. Eight studies examined the risk of placental abruption/infarction in the presence of vitamin B12 or folate deficiency, or hyperhomocyst(e)inaemia. Folate deficiency was a prominent risk factor for placental abruption/infarction among four studies, though not statistically significant (pooled odds ratio 25.9, 95 per cent CI 0.9-736.3). Hyperhomocyst(e)inaemia was also associated with placental abruption/infarction both without (pooled odds ratio 5.3, 95 per cent CI 1.8-15.9) and with methionine loading (pooled odds ratio 4.2, 95 per cent CI 1.2-15.0), as was the homozygous state for methylenetetrahydrofolate reductase (pooled odds ratio 2.3, 95 per cent CI 1.1-4.9). Vitamin B12 deficiency was not a demonstrable risk factor. Eight studies examined blood levels among women with spontaneous abortion or recurrent pregnancy loss. The pooled odds ratios were 3.4 (95 per cent CI 1.2-9.9) for folate deficiency, 3.7 (95 per cent CI 0.96-16.5) for hyperhomocyst(e)inaemia following methionine challenge, and 3.3 (95 per cent CI 1.2-9.2) for the methylenetetrahydrofolate reductase mutation. Five case control studies examined the relationship between pre-eclampsia and abnormal levels of vitamin B12, folate, homocyst(e)ine or methylenetetrahydrofolate reductase. Folate deficiency was not an associated risk factor (odds ratio 1.2, 95 per cent CI 0.5-2.7), but hyper-homocyst(e)inaemia was (pooled odds ratio 20.9, 95 per cent CI 3.6-121.6). Similarly, homozygosity for the methylenetetrahydrofolate reductase thermolabile variant was associated with a moderate risk of preeclampsia (odds ratio 2.6, 95 per cent CI 1.4-5.1). Some pooled data were associated with significant statistical heterogeneity, however. There is a general agreement among several observational studies that folate deficiency, hyperhomocyst(e)inaemia and homozygosity for the methylenetetrahydrofolate reductase thermolabile variant are probable risk factors for placenta-mediated diseases, such as pre-eclampsia, spontaneous abortion and placental abruption. Vitamin B12 deficiency is less well defined as an important risk factor. Due to the limited quality of these data, including insufficient matching of cases with controls, and possible laboratory measurement bias relating to pregnancy, prospective studies are needed to confirm these findings and guide future preventative and therapeutic research. PMID- 10452906 TI - Trophoblast deportation in human pregnancy--its relevance for pre-eclampsia. AB - The maternal syndrome of pre-eclampsia is thought to result from endothelial cell damage caused by a circulating factor derived from the placenta. This study investigates the hypothesis that trophoblast deportation may be part of the process by which this factor enters the maternal circulation. The nature and incidence of trophoblast deportation was studied in uterine vein and peripheral blood taken from normal and pre-eclamptic women at caesarean section. Trophoblasts were enriched using immunomagnetic beads to deplete leucocytes and labelled with trophoblast-specific monoclonal antibodies. Syncytiotrophoblast, cytotrophoblast, cytotrophoblast clumps and anucleate trophoblast cells were found in uterine vein blood. Cytotrophoblast cells were found to be shed less frequently than syncytiotrophoblast and the majority were probably villous in origin. Trophoblasts were found in the uterine vein blood of normal pregnant women with higher levels in pre-eclampsia. However, trophoblasts were rarely found in the peripheral circulation. There was no correlation between trophoblast numbers and either the severity of the disease, the extent of placental pathology or the inhibitory effect of uterine and peripheral vein plasma on endothelial growth in vitro. Thus, it is speculated that increased trophoblast deportation in pre-eclampsia is secondary to the structural and functional changes occurring in the placenta, rather than directly linked with the circulating endothelial cell damaging factor in pre-eclampsia. PMID- 10452907 TI - Glutathione and glutathione-related enzymes in decidua and placenta of controls and women with pre-eclampsia. AB - Pre-eclampsia is a major complication of pregnancy with high morbidity and mortality rates. The aetiology is still unclear but impaired detoxification or enhanced levels of reactive (oxygen) metabolites may contribute to the development or maintenance of pre-eclampsia. Glutathione and glutathione-related enzymes, as one of the major detoxificating and free-radical scavenging systems, may play a role in controlling the disease. Seventeen normotensive pregnant women and 24 pre-eclamptic women were investigated prospectively with respect to placental and decidual levels of total glutathione (GSH), glutathione S transferase activity (GST), selenium-dependent glutathione peroxidase (SeGPX) and total glutathione peroxidase activity (TGPX, both selenium- and non-selenium dependent GPX). Decidual levels of glutathione and related enzymes were compared with placental levels, and the investigated parameters in pre-eclampsia were compared with those in normotensive pregnancy by the Mann-Whitney U -test. Clinical data were correlated with biochemical parameters by Spearman's correlation test. Glutathione levels were significantly higher in decidua as compared with placenta. Glutathione levels were elevated in pre-eclampsia and HELLP (haemolysis, elevated liver enzymes, low platelets) as compared to normotensive pregnancy for decidua and in the placenta of patients with pre eclampsia only. Glutathione S-transferase activity was not different between the two groups. In the placenta of patients with pre-eclampsia+HELLP, total glutathione peroxidase activity was elevated versus controls. Selenium-dependent glutathione peroxidase activity was higher in decidua versus placenta and in decidua of pre-eclamptic versus control subjects. Enhanced glutathione concentrations and glutathione peroxidase activities were often found in placenta and decidua in pre-eclampsia, probably as a compensatory mechanism to prevent further damage by peroxides, (oxygen) radicals or other toxins in the placenta or in the feto-placental interface. PMID- 10452908 TI - Characteristic growth of human choriocarcinoma xenografts in nude mice. AB - In order to investigate the mechanisms and regulation of trophoblast vessel invasion, we established a model using malignant trophoblast cells grown in nude mice. The human choriocarcinoma cell line Jeg-3 established fast-growing tumours within 2 weeks after subcutaneous injection into nude mice. Interestingly, instead of neoangiogenesis the tumours were characterized by large blood-filled lacunae. Staining for hCG-beta and for mouse panendothelial antigen revealed that the majority of the lacunae were lined by trophoblast cells. The blood supply of the lacunae resulted from invasion of the choriocarcinoma cells into the host vessels at the junctional zone between tumour and mouse tissue. These large blood filled lacunae led to a much faster expansion of tumours with less necrotic areas compared to tumours of non-trophoblastic origin (Hec-1A and HeLa), which revealed neovascularization. Jeg-3 choriocarcinoma cells eroded host vessels and replaced the endothelial lining in a similar way to trophoblast cells during the establishment of a haemochorial placenta. This model can be useful in investigating the cell biological mechanisms of trophoblast vessel invasion and replacement of the endothelium by trophoblast cells. PMID- 10452909 TI - Annexin V inhibits phosphatidylserine-induced intrauterine growth restriction in mice. AB - To investigate the role of phosphatidylserine (PS) derived from the activated platelets in placental circulation, we established an artificial PS-induced intrauterine growth restriction (IUGR) model in mice and examined whether annexin V, a PS-binding anticoagulant protein, could prevent the development of IUGR in the animal experiments. The ICR mice were injected in the tail vein on days 8, 11 and 14 of pregnancy with filtered PS/phosphatidylcholine (PC) microvesicles (<0.3 microm in maximal diameter). The microvesicles have a procoagulant activity which was inhibited by annexin V in a dose-dependent fashion. The mice were killed on day 18 of pregnancy and the placental and fetal weights were measured. The placental tissue specimens were examined microscopically. PS/PC vesicles induced significant reductions in fetal weights compared with PC vesicles alone. The placental tissue revealed severe congestion with fibrin depositions although the lung and kidney tissue specimens showed minimal histological changes. PS/PC microvesicles with recombinant annexin V showed no significant reductions in fetal weights in mice with PS/PC vesicles alone. These results suggest that PS from the activated platelets induces IUGR by enhancing the coagulation cascade in the placental circulation and that annexin V from the villous trophoblast prevents the development of IUGR. PMID- 10452910 TI - Expression of thioredoxin and thioredoxin reductase in placentae of pregnant mice exposed to lipopolysaccharide. AB - We have previously shown that thioredoxin and thioredoxin reductase were immunohistochemically localized in cytotrophoblasts, decidua and stromal cells in the stem villi of human placenta and that the addition of exogenous thioredoxin and thioredoxin reductase to mitochondrial fractions from human placenta displayed a protective effect on fumarase activity against oxidative stress. In this study, to investigate further the roles of thioredoxin and thioredoxin reductase in protecting pregnancy against oxidative stress, we examined the effect of lipopolysaccharide (LPS), which induces a variety of cytokines and produces radical oxygen species, on the expression of thioredoxin and thioredoxin reductase in mouse placenta. We focused on the placental protective effect in the second trimester, when the onset of placental dysfunction might occasionally lead to a critical state for the fetus. Thus we analysed placentae from mice on day 13 of pregnancy at various time points after they were injected with LPS (50 microg/kg i.p.) or saline as a control. The expressions of thioredoxin and thioredoxin reductase were evaluated by Western blotting and immunohistochemistry. Western blot analysis revealed that LPS approximately quadrupled the expression of both thioredoxin and thioredoxin reductase in the placentae of pregnant mice. When both proteins were localized immunohistochemically, it was found that the decidua and the diploid trophoblasts in the basal zone were intensively stained. Furthermore, the expression of 4 hydroxy-2-nonenal (HNE)-modified proteins, which are markers of oxidative stress, was enhanced in placenta by LPS. Our study suggests that the induced thioredoxin and thioredoxin reductase might protect the placenta from the stress induced by LPS. PMID- 10452911 TI - Labour-associated expression of intercellular adhesion molecule-1 (ICAM-1) in placental endothelial cells indicates participation of immunological processes in parturition. AB - Inflammatory cytokines induce or upregulate de novo expression of cell adhesion molecules on endothelial and epithelial cells. In order to demonstrate inflammatory reactions within placental tissues in association with normal term as well as non-infection-induced preterm labour, the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and endothelial leucocyte adhesion molecule-1 (ELAM-1) was examined by immunohistochemical methods in both trophoblastic villi (n=123) and umbilical cord (n=61). As a result, ICAM-1 immunoreactivity was exclusively localized in the endothelial cells of the fetal vascular system, while VCAM-1 and ELAM-1 were not detected. Whereas ICAM-1 was not expressed in early pregnancy (9-12 weeks of gestation), it could be weakly detected at the end of pregnancy in cases of elective caesarean delivery in the absence of labour, and was significantly more strongly expressed in cases of vaginal delivery after spontaneous onset of normal term labour. Significantly increased immunoreactivity of ICAM-1 within umbilical cord tissues was also found in association with uncontrollable preterm labour in the absence of intrauterine infection which was excluded after histological examination of fetal membranes, umbilical cord and chorionic plate. We conclude that ICAM-1 expression in the endothelium of the fetal vascular system is associated with the presence of labour and reflects participation of immune inflammatory reactions in labour-promoting mechanisms. PMID- 10452912 TI - Expression of the 11beta-hydroxysteroid dehydrogenase types 1 and 2 proteins in human and baboon placental syncytiotrophoblast. AB - We have shown that the placenta, via metabolism of maternal cortisol and cortisone by the 11beta-hydroxysteroid dehydrogenase (11beta-HSD) enzymes types 1 and 2 in the syncytiotrophoblast, regulates the maturation of the fetal pituitary adrenocortical axis in the baboon. Because the timing and regulation of fetal adrenal development by fetal ACTH in the human seem to parallel that in the baboon, we propose that the placental 11beta-HSD-1 and -2 system also has a role in regulating the development of the fetal pituitary adrenocortical axis during human pregnancy. However, although the human placenta has been shown to express the 11beta-HSD-2, it remains to be determined unequivocally whether 11beta-HSD-1 protein is present in the human placental syncytiotrophoblast. To answer this question, enriched fractions of syncytiotrophoblast were prepared from human and baboon term placentae and proteins probed with polyclonal antibodies directed to amino acids 22-36 or 66-77 of human 11beta-HSD-1. The 11beta-HSD-1 was detected by Western blot analysis as a 32-kDa protein in human and baboon syncytiotrophoblast and as a 34-kDa protein in adult baboon liver. Localization of the 11beta-HSD-1 to the syncytiotrophoblast was confirmed by immunocytochemistry following antigen retrieval. These results show that both human and baboon placental syncytiotrophoblast expressed the 11beta-HSD-1, as well as the 11beta-HSD-2, proteins. Because 11beta-HSD-1 can function as a reductase, the expression of 11beta-HSD-1 in human syncytiotrophoblast would be consistent with the ability of this tissue to convert cortisone to cortisol and provide a means by which transplacental transport of cortisol could regulate the fetal pituitary adrenocortical axis in the human, as recently shown experimentally in the non-human primate baboon model. PMID- 10452913 TI - Presence of CLA-1 and HDL binding sites on syncytiotrophoblast brush border and basal plasma membranes of human placenta. AB - It is now known that rapid placental and fetal development is associated with elevated levels of circulating high density lipoprotein (HDL) in pregnant women. The main structure implicated in the maternal-fetal exchange is the syncytiotrophoblast, composed of a brush border membrane (BBM), facing the mother, and a basal plasma membrane (BPM), facing the fetus. In order to understand the mechanisms controlling the placental and fetal supplies of cholesterol, we purified both BBM and BPM and verified the presence of HDL binding sites in these membranes. Binding studies using(125)I-HDL(3)show a single affinity binding site on BPM which has a dissociation constant (K(d)) of 3.45+/ 0.43 microg protein/ml and a maximal binding capacity (B(max)) of 5.46+/-1.69 microg protein/mg membrane proteins. In BBM, we observed two affinity binding sites, one with a K(d)of 0.62+/-0.03 microg protein/ml and another one with a K(d)of 6.57+/-0.87 microg protein/ml. Their B(max)values were 0.54+/-0.11 and 2.34+/-0.39 microg of HDL(3)/mg membrane proteins, respectively. CLA-1, a putative HDL-receptor of 85 kDa, was detected on both BPM and BBM, together with two apo A-l binding sites of 110 and 96 kDa on BPM and BBM, respectively. These results provide further evidence that human placenta possesses specific sites for HDL binding, underlining the important role of maternal HDL in the transfer of cholesterol from the maternal circulation to the placenta and the fetus. PMID- 10452914 TI - Glucose production by the human placenta in vivo. AB - The human placenta transports glucose by facilitated diffusion down a concentration gradient from mother to fetus. It has previously been considered incapable of glucose synthesis. However, recent work has demonstrated the presence in placental tissue of glucose-6-phosphatase, which is required for the final step in the synthesis of glucose. Following continuous intravenous infusion into the maternal circulation of the stable isotope, 6,6-(2)H(2)glucose, during elective caesarean section, we have observed isotope dilution in the umbilical vein, without further dilution in the umbilical artery. Using a mathematical model containing maternal, placental and fetal compartments, the data were compatible with the release of glucose by the placenta. We conclude that the human placenta at term can produce glucose. PMID- 10452915 TI - Human carboxylesterases in term placentae: enzymatic characterization, molecular cloning and evidence for the existence of multiple forms. AB - The placenta is a temporary organ that is known to metabolize numerous endogenous and xenobiotic compounds. Carboxylesterases represent a family of enzymes which hydrolyse a variety of esters, amides and thioesters. Many studies have demonstrated that carboxylesterases are widely distributed among mammalian tissues, but little is known about these enzymes in the placenta. The present study was conducted to establish the kinetic parameters of placental carboxylesterases toward several p -nitrophenol and 1-naphthol esters, and to establish the molecular basis for these enzymes in the placenta. The enzymatic rate of the hydrolysis of 1-naphthylacetate and carboxylic esters of p nitrophenol as a function of substrate concentration (0.01-1.00 m m) was examined with human placental microsomes pooled from six placentae. Data from these studies yielded a linear Lineweaver-Burk plot with each substrate examined. K(m)values for these substrates ranged from 92 to 370 microm, and V(max)values ranged from 85 to 170 nmol/mg/min. These results suggest that each substrate is hydrolysed by a single enzyme, or enzymes that are kinetically indistinguishable, or that one of them is dominant. Microsomes from all individual placentae contained esterase activity toward all four substrates, and exhibited a one- to three-fold variation. The activity toward p -nitrophenylacetate correlated well with the activity toward 1-naphthylacetate (r(2)=0.957). In contrast, the activity toward p -nitrophenylbutyrate correlated poorly with the activity toward 1-naphthylacetate (r(2)=0.121). These results suggest that placental microsomes have more than one carboxylesterase activity. Screening of a placental cDNA library with gene-trapping hybridization resulted in the isolation of three distinct cDNAs, designated PCE-1, PCE-2 and PCE-3. PCE-1 and PCE-2 have a significant sequence identity (approx 99 per cent) with liver carboxylesterases hCE and hCE-2, respectively. PCE-3 has a 96 per cent sequence identity with hCE but only at the first 874 nucleotide of the 5' end. The rest of the 1396 nucleotides of the 3' end exhibit no significant sequence identity with any known mammalian carboxylesterases. A probe derived from the 3' end of PCE-3 detected an approx 2.2 kb messenger transcript, the size of a regular carboxylesterase. However, the entire PCE-3 cDNA has multiple internal stop codons and encodes only 269 amino acids; half the size of a regular carboxylesterase. Northern blotting experiments detected the transcripts coding for PCE-1, PCE-2 or PCE-3 in all placentae, and the levels of these messengers showed an approx six-fold individual variation. Placenta 6 had the highest activity toward all four substrates, and highest levels of the messengers for PCE-1, PCE-2 and PCE-3. In contrast, placenta 1 had relatively high levels of messengers for PCE-1 and PCE 2, but the activity toward these four substrates was only moderate. These results suggest that a discrepancy between the messenger level and the enzyme protein exists or that there are other as yet unidentified carboxylesterase(s) in the placenta which contribute to the hydrolytic activity. Carboxylesterases are known to involve the detoxication and metabolic activation of various drugs, environmental toxicants and carcinogens. Therefore, placental carboxylesterases have both pharmacological and toxicological significance in the development of the fetus. PMID- 10452916 TI - Parallel Doppler assessment yolk sac and intervillous circulation in normal pregnancy and missed abortion. PMID- 10452917 TI - Reply to Dr jauniaux PMID- 10452918 TI - Best practice guidelines for the management of women with epilepsy. The Women with Epilepsy Guidelines Development Group. AB - Clinical guidelines for the treatment of epilepsy have been published. A statement on management issues for women with epilepsy has recently been produced by the American Academy of Neurology which has raised awareness of the issues facing women with epilepsy. The communication presented here aims to review current literature on specific issues relating to women with epilepsy, and proposes graded recommendations for its management within a UK health care framework. PMID- 10452919 TI - Serum prolactin levels after seizure and syncopal attacks. AB - Loss of consciousness and falling are the key features of syncope. Common accompaniments include tonic and myoclonic muscle activity, eye deviations, automatisms, vocalizations and hallucinations that may render the distinction from epileptic seizures difficult. The frequently increased levels of serum prolactin (SPRL) were observed immediately after generalized and complex partial seizures. Presumably, the hormone release is caused by the propagation of epileptic activity, usually from the temporal lobe to the hypothalamic pituitary axis. Numerous reports have demonstrated that the post-ictal SPRL level may be used to differentiate between epileptic and syncopal, non-epileptic attacks. In order to confirm the hypothesis, the SPRL levels were measured in patients with complex partial seizures (CPS) and patients with vaso-vagal syncopal attacks (VVS). The SPRL levels were prospectively measured for each patient as soon as possible after the event (within 1 hour), then 1 hour after the first determination and finally blood was sampled 24 hours later. During the study period (18 months), 18 patients with CPS and 15 patients with VVS were investigated in total. The mean values of SPRL levels in both groups were increased immediately after the event (CPS group: 1142 +/- 305 mIU/l; VVS group: 874 +/- 208 mIU/l). The elevated SPRL levels were found in 14 (78%) patients immediately after CPS and in 9 (60%) patients immediately after VVS. After examining the results of the present study we conclude that the elevated serum prolactin level after an epileptic attack is of no significant value in differential diagnosis between epileptic and vaso-vagal syncopal attacks. PMID- 10452920 TI - Are we overusing the diagnosis of psychogenic non-epileptic events? AB - In order to determine how often results of video/EEG (V-EEG) studies may change the clinical diagnosis of paroxysmal events, we prospectively studied 100 consecutive patients (75 females, 25 males) admitted for diagnosis of recurrent paroxysmal spells. The presumed diagnosis of the referring physician was obtained. Episodes were classified as epileptic seizures (ES), psychogenic non epileptic events (PNEE), or physiologic non-epileptic events (PhysNEE). Eighty seven patients had diagnostic events. A final diagnosis of ES was made in 21 patients, PNEE in 39, PNEE + ES in 20, and PhysNEE in seven. All PhysNEE were unsuspected. ES were misdiagnosed as PNEE more frequently than the reverse (57% vs. 12%, P < 0.001). Among the 64 patients with recorded events who had been suspected of having PNEE, 14 (21.9%) were misdiagnosed: two had PhysNEE and 12 (18.75%) had ES. Among the 23 patients with recorded events who were thought to have ES, 12 (39.1%) were misdiagnosed: seven had PNEE, five PhysNEE. V-EEG changed the clinical diagnosis in 29.8% of the patients with recorded events. Our data suggests that clinicians have become more aware of PNEE since the advent of V-EEG and have little problem recognizing them. However, they may be more prone to make a false-positive diagnosis of PNEE in ES with some atypical features. At this point, efforts should be channeled to better training in the proper recognition of ES that mimic PNEE. PMID- 10452921 TI - Self-assessment of well-being in a group of children with epilepsy. AB - Epilepsy is common in childhood, the prevalence being about five per 1000 children. The purpose of this study was to assess well-being in children with controlled epilepsy (but did not include those with obvious neurodeficits such as mental retardation or cerebral palsy) and compare them with age-matched healthy children. The patient group comprised of 31 children, 12 boys and 19 girls, whereas the control population group consisted of 342 children, 176 boys and 166 girls who were all in good health. All children involved in the study were aged between 9-13 years. A questionnaire was distributed to the children to complete. It consisted of 39 bipolar adjectives and a visual analogue scale was employed. The results show that the group of children with controlled epilepsy did not differ significantly from the age-matched control group. There was no significant difference between the sexes except for the dimension of vitality, where the boys scored better than the girls. Thus the well-being of children with controlled epilepsy seems to be similar to that of children from a control population. The psychometric properties of the instrument were also assessed. An assessment of well-being in children with intractable epilepsy, using a similar approach, is in progress. PMID- 10452922 TI - Acute psychotic symptoms induced by topiramate. AB - The incidence of psychosis during clinical trials of topiramate was 0.8%, not significantly different from the rate for placebo or reported rates of psychosis in patients with refractory epilepsy. We observed psychotic symptoms in five patients soon after initiation of topiramate therapy. We performed a retrospective chart review of the first 80 patients who began on topiramate after approval for clinical use, between January and April 1997. Symptoms suggestive of psychosis, including hallucinations and delusions, were sought for analysis. Cognitive effects such as psychomotor slowing, confusion, and somnolence were not included. Five patients developed definite psychotic symptoms 2 to 46 days after beginning topiramate. Dosages at symptom onset were 50-400 mg/day. Symptoms included paranoid delusions in four patients and auditory hallucinations in three. Symptoms of psychosis and other psychiatric symptoms resolved quickly with discontinuation of topiramate in three patients, dose reduction from 300 to 200 mg/day in one and with inpatient treatment and neuroleptics in another. One patient had a history of auditory hallucinations, one of aggressive and suicidal thoughts, but three had no significant psychiatric history. Physicians should be aware of the possibility of psychotic symptoms, even in patients without a previous psychiatric history, when prescribing topiramate. Symptoms resolve quickly with discontinuation. PMID- 10452923 TI - Dopey's seizure. AB - Angelman syndrome is a neurogenetic condition namely characterized by developmental delay, virtual absence of expressive verbal language, peculiar organization of movement, seizures and happy demeanor. This syndrome has been recognized since 1965, but it seems that Walt Disney presented an original depiction of it in his first full-length animated film, including myoclonic jerks and an apparently generalized tonic-clonic seizure. PMID- 10452924 TI - An analysis of neuropsychological change scores following selective temporal resection of the non-dominant temporal lobe. AB - Reliable change in neuropsychological test scores was examined in patients undergoing right-sided, selective temporal resections for the relief of intractable epilepsy. Measures were taken prior to surgery, 1-month post operatively and 1-year post-operatively. Non-verbal memory performance was more robustly measured than in previous studies. Results failed to replicate previous studies which report verbal memory deficits even following right-sided surgery. No strong evidence of a material-specific, non-verbal memory deficit was found on commonly used tests of non-verbal memory. The majority of patients failed to show reliable decline in performance following surgery indicating that fears of post operative memory decline may be unfounded. PMID- 10452925 TI - Benign rolandic epilepsy: neuropsychological findings. AB - Benign rolandic epilepsy (BRE) is a partial idiopathic epilepsy of childhood presenting with a nocturnal seizure and with a typical EEG showing centrotemporal spike and multifocal or generalized sharp slow waves. Although normal neurological and intellectual development are expected in BRE, it is not infrequent to detect subtle defects in neuropsychological functions and neuromotor development. This study included 20 cases of BRE diagnosed according to the criteria of ILAE. The patients underwent several tests of neuropsychological functions as well as detailed neurological examination and the results were compared statistically to normal controls. In the patient group, a family history of language delay or learning disability (P < 0.005), presence of consanguinity (P < 0. 05), dyspraxia in the lower extremities (to imitation) (P < 0.05), difficulties in go-no-go test (P < 0.001), as well as some problems related to language such as dysprosody (P = 0.05), minor motor deficits in the left (P < 0.05) and right upper extremity (P < 0.05) were significantly more frequent compared to the control group. One should be rather guarded against the prognosis in BRE with respect to the higher cortical functions and neurodevelopmental problems. PMID- 10452926 TI - Carbamazepine, hepatotoxicity, organic solvents, and paints. AB - Hepatotoxicity secondary to carbamazepine is a serious condition which can be fatal. However, other concomitant medications or environmental factors may be the offending agents. In this case report, hepatotoxicity secondary to organic solvents and paints is described. PMID- 10452927 TI - Failure of periconceptual folic acid to prevent a neural tube defect in the offspring of a mother taking sodium valproate. PMID- 10452928 TI - Enough intervention AB - Poetry is meant to express the emotions and thoughts often found difficult to state or even to put into prose. This clinician has dedicated his professional life to the rational treatment of epilepsy - for he understands only too well the consequences of varied interventions over a forty-year course of his own epilepsy. This poem is meant to remind other clinicians that quality of life may not always mean total seizure control. Copyright 1999 BEA Trading Ltd. PMID- 10452929 TI - Principles of successful cancer screening. AB - Screening for cancer is the application of various tests to apparently healthy individuals in order to identify who among them has occult disease, which may be either invasive disease or a precursor lesion. For any given cancer site, the potential of screening to reduce morbidity and mortality is based on well-defined criteria for the evaluation of screening effectiveness and on acceptable performance to be realized in the average community setting. Screening programs are most successful when they are organized into a system that leads to high rates of participation, high quality, and constant surveillance and evaluation. If the elements that contribute to a successful screening program are not well organized and integrated, then the fullest potential of screening will not be realized. PMID- 10452930 TI - Defining responsibility for screening. AB - Patients commonly receive medical care from multiple providers and confusion as to who is responsible for cancer screening undoubtedly contributes to inadequate recommendations. Effective screening requires successful implementation of a series of steps that begin with the initial discussion of a screening test and proceed through obtaining results and instituting appropriate follow-up. Clear definition of generalist and specialist physician roles are necessary to optimally screen the public. This article explores the differences in how generalists and specialists approach screening, describes models of care that facilitate shared responsibility for screening, and suggests strategies on how to improve communication between physicians to maximize screening performance. PMID- 10452931 TI - Issues in cytologic screening and evaluation. AB - A variety of cytologic specimens may be useful in the screening and evaluation of patients for neoplastic processes. Fine needle aspiration cytology is an example of a cost-effective procedure for mass lesions and may provide a definitive diagnosis for treatment purposes. The sensitivity of a single cytologic procedure varies from 70% to more than 90% depending on specimen type and procurement methods. Communication with the laboratory, including provision of clinical information, is important to ensure proper specimen submission and handling and will also clarify cytologic terminology and appropriate follow-up measures. PMID- 10452933 TI - Breast cancer screening: success amid conflict. AB - In randomized trials, screening mammography has been shown to reduce breast cancer mortality. There has been controversy, however, about the segment of the population that should be screened and the frequency at which screening should occur. This article details these controversial issues. The benefits that have been realized from mammography screening in overall breast cancer mortality and diagnosis of earlier stage cancers are also reviewed. PMID- 10452932 TI - The potential of molecular screening. AB - Clonal genetic alterations such as tumor suppressor, gene mutations, or microsatellite instability are a hallmark of human cancer. These genetic alterations can be detected in clinical samples using DNA amplification techniques and used for early detection of cancer. This article reviews molecular based approaches for screening of malignancies commonly encountered by the practicing general surgeon. PMID- 10452934 TI - Colorectal cancer screening: new opportunities. AB - Colorectal cancer is the third most common cancer among men and women in the United States, and the third leading cause of cancer death. Strategies currently available to screen for colorectal cancer include fecal occult blood tests, sigmoidoscopy, or both tests used in combination. Colonoscopy and double contrast barium enema are potentially preferable options because they offer improved sensitivity over currently available tests, but the feasibility of these tests for population screening remains in doubt. Future opportunities for screening include focusing special efforts to deliver screening to higher risk individuals based on family history or age and the use of molecular or computer-aided radiographic techniques as alternatives to colonoscopy. PMID- 10452935 TI - Prostate cancer: A challenge for screening. AB - Prostate cancer is the second leading cause of cancer related death in men over the age of 50 in the United States today. The serum prostate specific antigen has the highest predictive value of any diagnostic test for prostate cancer. Highly screened communities have realized an impressive downward stage migration, younger average age at detection, and an improved five-year survival rate. These improvements are necessary components for any successful screening program. PMID- 10452936 TI - Screening for gynecologic malignancies: A continuing responsibility. AB - Screening for cervical cancer with the Papanicolaou cervical smear has resulted in a decline in incidence and mortality from cervical cancer. Targeting the unscreened population is the next challenge to reduce the incidence of this disease further. Currently, there are no available screening modalities for endometrial or ovarian cancer. Breakthroughs in molecular genetics may result in screening tests for ovarian cancer. PMID- 10452937 TI - Screening for cancers of the head and neck: addressing the problem. AB - More than 90% of upper aerodigestive tract (UADT) cancers occur in people with specific lifestyle risks, including tobacco and alcohol use. More than 90% of tumors occur in easily examined parts of the head and neck, therefore, there is the possibility of identifying the vast majority of patients through selective screening. Physicians should keep in mind that the mucosa's sojourn from visually suspicious (and possibly malignant) tissue is most likely less than two years, and frequent examination of asymptomatic patients is necessary. When patients wait to bring symptoms to medical attention, their cancers will be advanced 60% of the time when the chance of cure is less than 30%. Given the difficulty of implementing regular examinations in a poorly compliant, high risk population, genetic and molecular screening tools may allow very high risk individuals to be identified. PMID- 10452938 TI - Skin cancer screening: A growing need. AB - Skin cancer is the most frequently diagnosed cancer in the United States. Melanoma is the major cause of deaths due to skin cancer, however, squamous cell carcinoma and basal cell carcinoma account for considerable morbidity also. All three types are potentially curable if diagnosed at an early stage. Visual examination of the skin by a trained observer is a simple and effective screening tool which may be utilized in a multitude of settings. Patient education and self examination augment the impact of office-based screening and mass screening programs. PMID- 10452939 TI - Screening for lung cancer: An evidence-based synthesis. AB - At present, screening for lung cancer is not recommended; however, consideration of the evidence on the basis of conventional screening criteria, including burden of disease, detectable preclinical phase, accuracy, and acceptability, suggests that screening is likely to be highly beneficial. The most important question relates to effectiveness and cost-effectiveness, which in turn is paradigm dependent. An evidence-based synthesis supports the conclusion that chest radiographs deserve to become the community standard for those at high risk for lung cancer. PMID- 10452940 TI - Unlocking the mechanisms of transcription factor YY1: are chromatin modifying enzymes the key? AB - The transcription factor YY1 is a complex protein that is involved in repressing and activating a diverse number of promoters. Numerous studies have attempted to understand how this one factor can act both as a repressor and an activator in such a wide set of different contexts. The fact that YY1 interacts with a number of key regulatory proteins (e.g. TBP, TFIIB, TAFII55, Sp1, and E1A) has suggested that these interactions are important for determining which particular function of YY1 is displayed at a specific promoter. Two groups of proteins, previously known to function as corepressors and coactivators, that now seem likely to modulate YY1's functions, are the histone deacetylases (HDAC) and histone acetyltransferases (HAT). These two groups of enzymes modify histones, and this modification is proposed to alter chromatin structure. Acetylated histones are typically localized to active chromatin while deacetylated histones colocalize with transcriptionally inactive chromatin. When these enzymes are directed to a promoter through a DNA binding factor such as YY1, that promoter can be activated or repressed. This review will discuss the recent work dealing with the different proteins that interact with YY1, with particular emphasis on ones that modify chromatin, and how they could be involved in regulating YY1's activities. PMID- 10452941 TI - Conserved cAMP responsive element and core promoter complex are critical for specificity of the distal T-cell receptor beta chain enhancer for its native promoter. AB - The Vbeta 8.1 promoter is regulated by T-cell-specific and ubiquitous transcription factors, which bind immediately upstream of and inside the core promoter region. The various Vbeta promoters contain two conserved elements, a cAMP responsive element (CRE) located upstream of the core promoter and a basal initiator flanked by two regulatory motifs. Here we have studied the interplay between the distal enhancer and its native promoter. We show that the remote enhancer acts specifically through its native promoter. Specific enhancer promoter interplay is mediated through the conserved regions of the Vbeta promoters. Importantly, the conserved CRE serves as a functional recognition element for the enhancer whereas it barely contributes to promoter activity. The other conserved regions surrounding the initiation site are critical for activators that bind at and function through the core promoter region and thereby regulate both promoter and enhancer activity. The enhancer is highly sensitive to E1A-12S, which represses both general and specific enhancer activities. Enhancer activity and promoter-enhancer specificity is, at least in part, mediated by the coactivators CBP/p300. PMID- 10452942 TI - Organization and expression of mouse nm23-M1 gene. Comparison with nm23-M2 expression. AB - Nm23 is a gene family encoding different isoforms of the nucleotide diphosphate kinase (NDPK), an enzyme involved in the synthesis of nucleoside triphosphates. In the present study, the organization and expression of the nm23-M1 gene encoding the mouse NDPKA isoform are described. This gene is about 10kb long and composed of five exons. The organization and the exon-intron boundaries are strictly conserved as compared to the human and rat related genes. The gene promoter region did not exhibit any consensus TATA box, SP1 binding element or Inr sequence. By contrast, TCF-1/LEF-1 binding elements and Pit-1 consensus sequence were present. Northern blotting and in situ hybridization methods were carried out in adult and 18.5 days post-coitum (dpc) mouse embryo, respectively. They showed tissue-specific expression of nm23-M1 transcripts, despite housekeeping gene promoter features. The strongest signals were detected in the nervous system, sensory organs and embryonic thymus. In contrast nm23-M2 mRNA was shown to be more widely expressed.The relationship between nm23-M1 gene tissue specific expression and the putative binding element of the promoter region is discussed. PMID- 10452943 TI - The TATA-less rat GAD65 promoter can be activated by Sp1 through non-consensus elements. AB - Glutamic acid decarboxylase (GAD) 65 is one of two homologous proteins responsible for the synthesis of gamma-aminobutyric acid, the most ubiquitous inhibitory neurotransmitter. In order to characterize the DNA elements responsible for controlling GAD65 expression, we cloned the 5' flanking region of the rat GAD65 gene. A major, proximal and a minor, distal region of transcription initiation were located by RACE experiments. Sequence analysis revealed that the initiation sites are located within a region devoid of TATA boxes. We investigated the functional organization of the promoter by measuring the ability of 5' deletion mutants to drive the expression of a luciferase reporter gene. The major promoter was found to be located in the region encompassing the 100bp immediately upstream of the proximal transcription initiation site. A number of near consensus GC boxes and initiator elements are found in this region, but gel shift assays suggest that they play only a minor role in transcription initiation. However, gel-shift assays and reporter gene assays suggest that Sp1 can bind to a region devoid of consensus Sp1 binding sites. PMID- 10452944 TI - Drosophila melanogaster is polymorphic for a specific repeated (CATA) sequence in the regulatory region of hsp23. AB - To identify sequence variation associated with a selection response for heat tolerance in Drosophila melanogaster, we sequenced 1400bp of the heat shock protein 23 gene (hsp23) promoter region in four heat-selected and two control lines. The region was found to be variable for a specific (CATA) repeated sequence, and the sequence CTT seems to be a hot spot for mutation. The repeated tetranucleotide sequence was located in several short repeats scattered throughout the entire region. Similar variable repeats are also located downstream the of hsp23 gene in the intergenic region between hsp23 and hsp27. We detected nine different hsp23 alleles. Their frequencies in the selection and control lines seemed to be mainly determined by genetic drift. The function of the CATA repeats is not yet known, though these regions have homology to SAR elements located in the intergenic region between two hsp70 genes, suggesting a similar function. PMID- 10452945 TI - Cloning and expression analysis of the sheep ceruloplasmin cDNA. AB - The cDNA encoding sheep ceruloplasmin (sCP) was isolated from a sheep liver cDNA library. The cDNA contig was 3530 nucleotides in length and encoded a protein of 1048 amino acids. The deduced amino acid sequence showed a high degree of conservation (87%) when compared to the human ceruloplasmin (hCP) sequence. Northern blot analysis of sheep tissue revealed that the sheep ceruloplasmin gene (sCP) was expressed primarily in the liver, but low levels of mRNA were detected in the hypothalamus, spleen and uterus. No sCP mRNA was detected in the cortex, heart, intestine or kidney. Expression was not significantly affected by hepatic copper content. Northern blot analysis of sheep liver during development demonstrated little sCP expression during fetal life, but significant levels of mRNA were observed after birth. Significantly, the developmental expression pattern of sCP was closely correlated with that of the sheep Wilson disease gene (sATP7B), suggesting that the expression of the two genes may be coordinated to ensure that copper is supplied to apoceruloplasmin. Overall, the structure and expression of sCP appeared similar to other mammals, suggesting that abnormalities in CP were not responsible for the unusual sheep copper phenotype. PMID- 10452946 TI - Cloning and characterization of the alternative promoter regions of the human LIMK2 gene responsible for alternative transcripts with tissue-specific expression. AB - We previously isolated the human LIMK2 gene and identified two alternative transcripts, LIMK2a and LIMK2b, which were differentially regulated in a tissue specific manner. To examine this differential tissue-specific expression in detail, an RNase protection assay was performed, which demonstrated three expression patterns of the LIMK2 isoforms. In digestive organs, the LIMK2a transcripts were preferentially expressed in fetal and adult tissues; in brain and lung, the LIMK2a transcript was predominantly expressed only in fetal tissue, and in placenta, the LIMK2b transcript was expressed more abundantly than that of LIMK2a. To further investigate this mechanism and the transcription factors involved, we isolated the two distinct 5' upstream regions from the phage genomic library and found that both LIMK2a and 2b promoters have a single major transcription initiation site and the characteristics of a TATA-less promoter. A luciferase reporter assay of the transcriptional activity revealed positive as well as negative regulatory regions within both promoters. The co-transfection assay suggested that the MZF-1 might regulate the expression of the LIMK2 isoforms in a different manner. The RORalpha1 might also be involved in the transcriptional regulation of the LIMK2b isoform. The genomic structure of the LIMK2 gene was also determined. These findings should lead to a better understanding of the possibly diverse functions of the LIMK family. PMID- 10452947 TI - Identification of two transcripts of canine, feline, and porcine interleukin-1 alpha. AB - Reverse transcription-polymerase chain reaction (RT-PCR) with interleukin-1alpha (IL-1alpha)-specific primers using total RNA from lipopolysaccharide (LPS) stimulated lung macrophages resulted in the amplification of two distinct cDNA fragments. Cloning and sequencing of the canine and feline fragments revealed that, except for the absence of a specific 174 nucleotide sequence, the short and the long transcripts were identical. The in-frame 174 nucleotide deletion corresponds to exon 5 of the human and murine IL-1alpha gene, which encodes the cleavage site for calpain, a protein necessary for the processing of the IL 1alpha precursor into mature IL-1alpha. The two transcripts were found in the dog, cat and pig; analysis by RT-PCR, Southern and Northern blot hybridization showed no expression of the shorter IL-1alpha mRNA in equine or bovine macrophages. Expression of the two canine IL-1alpha transcripts was also detected in synovial membranes and was coordinately up-regulated in response to Borrelia burgdorferi infection under both in-vitro and in-vivo conditions. PMID- 10452948 TI - Small subfamily of olfactory receptor genes: structural features, expression pattern and genomic organization. AB - Olfactory receptors of the OR37 subfamily are characterized by distinct sequence features and are expressed in neurons segregated in a restricted area of the olfactory epithelium. In the present study, we have characterized the complement of OR37-like genes in the mouse. Five OR37-like genes were identified. They reside within only 60kb of DNA on chromosome 4. About 70kb distant from this cluster, two additional olfactory receptor genes are located, which are members of distinct receptor subfamilies. Phylogenetic analysis demonstrated that the two physically linked receptors are closely related to the OR37 subfamily. Studies of gene expression showed that both genes are also expressed in clustered neuron populations located in the typical OR37 region of the epithelium. These data suggest the involvement of locus-dependent mechanisms for the spatial control of OR gene expression. PMID- 10452949 TI - Cloning and characterization of the gene encoding Aspergillus nidulans DNA topoisomerase II. AB - We have determined the complete nucleotide sequence of a 5544bp genomic DNA fragment from Aspergillus nidulans that encodes DNA topoisomerase II (topo II). It contains a single open reading frame of 4740bp that codes for 1579 amino acid residues with a molecular weight of 178kDa; when expressed in Escherichia coli and Saccharomyces cerevisiae the molecular weight was 180kDa. The gene (TOP2) is divided into three exons. Two introns, 54bp and 60bp in length, are located at nucleotide positions 187 and 3214 respectively. Comparison of the deduced amino acid sequence with other eukaryotic topo II sequences showed a higher degree of identity with other fungal enzymes than the human topo IIalpha. One of monoclonal antibodies raised against human topo II, 6H8, can cross-react with Aspergillus topo II. PMID- 10452950 TI - Organization and transcriptional regulation of Drosophila Na(+), K(+)-ATPase beta subunit genes: Nrv1 and Nrv2. AB - Drosophila melanogaster has two Na(+),K(+)-ATPase beta subunit genes (Nervana 1 and 2; Nrv), with tissue-specific expression patterns. Nrv1 produces a single beta subunit isoform expressed primarily in muscle tissue, whereas Nrv2 codes for two different isoforms (2.1 and 2.2) expressed in the nervous system. We have determined the complete molecular genomic organization for both Nrv genes. Only 3kb of DNA separate the 3' end of Nrv2 from Nrv1. The cDNAs from all three forms of Nrv have been mapped onto the genomic structure and all intron-exon junctions have been confirmed by direct sequencing. The genomic DNA positioned in the 5' flanking region of each Nrv gene has also been tested for tissue-specific transcriptional regulatory activity. P-element transformation vectors were constructed, which contained either 7.7kb of Nrv2 or 3.5kb Nrv1 5' flanking DNA driving expression of a lacZ reporter gene. Multiple transgenic Drosophila lines were established for each construct and analyzed for their beta-galactosidase expression pattern. The tissue-specific expression of each Nrv gene is independently regulated by the cis-element(s) present in the 5' flanking region. The Nrv2 5' flanking DNA directs expression exclusively to the nervous system, whereas Nrv1 5' flanking DNA directs expression primarily in muscle tissue. PMID- 10452951 TI - Isolation and characterisation of the retinoic acid receptor-alpha gene in the Japanese pufferfish, F. rubripes. AB - Nuclear hormone receptors (NRs) are ligand-inducible transcription factors that mediate critical functions in many species. The majority of novel NRs have hitherto been cloned from cDNA libraries by virtue of their homology to previously identified receptors. In this study, we validate a genomic DNA-based approach to isolating NRs by cloning the retinoic acid receptor-alpha (RARalpha) gene from the genome of the Japanese pufferfish, Fugu rubripes. The fRARalpha gene is more compact than its human and murine counterparts and demonstrates a highly conserved genomic organisation and amino acid sequence, generating two isoforms (fRARalpha1 and fRARalpha2) with divergent aminoterminal domains. In addition, a conserved regulatory element containing a retinoic acid response element was identified upstream of the fRARalpha2-specific exon, implying that retinoid induction of this isoform is evolutionarily conserved and critical to its function in vivo. We propose two uses for the Fugu genome in the study of NRs: the isolation of novel NRs that exhibit restricted spatio-temporal expression from genomic DNA and the identification of evolutionarily conserved promoter or intragenic regulatory DNA elements. PMID- 10452953 TI - Primary structure and features of the genome of the Lactobacillus gasseri temperate bacteriophage (phi)adh. AB - The complete DNA sequence of the Lactobacillus (Lb.) gasseri temperate phage (phi)adh was determined. The linear and double-stranded genome consists of 43.785bp with a G+C content of 35. 3% and 3' protruding cohesive ends of 12nt. Sixty-two possible ORFs were identified. On the basis of homology comparisons, some of them could be assigned to possible functions, such as a helicase, a nucleic acid polymerase and a protease. In a non-coding area of the (phi)adh genome, structural features of a potential replication origin were detected. After subcloning, this region was functional as a replicon in Lb. gasseri and Lactococcus lactis. N-terminal aa sequencing and electron microscopic analysis of intact and defective phage particles enabled the identification of two capsid protein genes. One of their products, the major head protein, seems to be processed on the posttranslational level. PMID- 10452952 TI - Molecular characterization and mapping of canine cGMP-phosphodiesterase delta subunit (PDE6D). AB - cGMP-phosphodiesterase (PDE) is composed of two catalytic (alpha and beta) and two identical inhibitory (gamma) subunits. The human gene (PDE6D) encoding a new subunit (delta) has been characterized and mapped to the long arm of chromosome 2 (HSA2q35-q36) where a new autosomal recessive retinitis pigmentosa (arRP) locus (RP26) has been localized. Characterization of the canine PDE6D shows the gene is about 4.2kb containing four exons interrupted by three introns; the size of the cDNA is 1059bp with an open reading frame (ORF) of 453bp. A single transcript of identical size (1.43kb) was detected in all tissues examined (liver, lung, spleen, kidney, heart, brain and retina), with the highest abundance in the retina. Canine PDE6D has been localized to canine radiation hybrid group 14-a, which extends conserved synteny between the dog, human chromosome 2q and mouse chromosome 1. The characterization of the canine PDE6D gene and its mapping provide important information for testing causal association of the gene with canine retinal degenerations, in particular rod-cone dysplasia 2 (rcd2) in collie dogs. This disease is characterized by abnormal retinal cGMP metabolism due to a deficiency in cGMP-PDE activity, yet the alpha, beta and gamma subunits of PDE have been excluded as candidate gene loci. PMID- 10452954 TI - Ras-mediated signaling pathway regulates the expression of a low-molecular-weight heat-shock protein in fission yeast. AB - In fission yeast, Schizosaccharomyces pombe, deficiency of ras1 gene causes an abnormal cell shape and abolishes mating ability. However, target genes of this signaling pathway are largely unknown because of the lack of an appropriate analysis system. To overcome this problem, we have started a novel project to categorize entire genes based on their expression levels under different growth conditions. Using this strategy, we screened genes whose expression levels were affected in the presence or absence of the ras1 gene product. For this purpose, we utilized high-density arrays of clones covering the entire genome of the fission yeast, and probed with labelled cDNA derived from various strains and growth conditions. Here, we demonstrate the detection of a low-molecular-weight heat-shock protein gene, hsp16, whose expression is very likely to be regulated by a ras-mediated signaling pathway, but not by the heat-shock response. PMID- 10452955 TI - Increase of collagen synthesis and deposition in the arachnoid and the dura following subarachnoid hemorrhage in the rat. AB - Arachnoidal fibrosis following subarachnoid hemorrhage (SAH) has been suggested to play a pathogenic role in the development of late post-hemorrhagic hydrocephalus in humans. The purpose of this study was to investigate the rate of collagen synthesis in the arachnoid and the dura in the rat under normal conditions and to study the time schedule and the localization of the increased collagen synthesis following an experimental SAH. We found that the activity of prolyl 4-hydroxylase, a key enzyme in collagen synthesis, was 3-fold higher in the dura than that in the arachnoid and was similar to the activity in the skin. We then induced SAH in rats by injecting autologous arterial blood into cisterna magna. After SAH, we observed an increase in prolyl 4-hydroxylase activity of the arachnoid and the dura at 1 week. At this time point the enzyme activity in both tissues was 1.7-1.8-fold compared to that in the controls and after this time point the activities declined but remained slightly elevated at least till week 4. The rate of collagen synthesis was measured in vitro by labeling the tissues with [(3)H]proline. The rate increased to be 1.7-fold at 1 to 2 weeks after the SAH in both of the tissues. Immunohistochemically we observed a deposition of type I collagen in the meninges at 3 weeks after the SAH. SAH is followed by a transient increase in the rate of collagen synthesis in the arachnoid and, surprisingly, also the dura. Increased synthesis also resulted in an accumulation of type I collagen in the meningeal tissue, suggesting that the meninges are a potential site for fibrosis. The time schedule of these biochemical and histological events suggest that meningeal fibrosis may be involved in the pathogenesis of late post-hemorrhagic hydrocephalus. PMID- 10452956 TI - Expression of human inositol monophosphatase suppresses galactose toxicity in Saccharomyces cerevisiae: possible implications in galactosemia. AB - A suppressor of galactose toxicity in a gal7 yeast strain (lacking galactose 1 phosphate uridyl transferase) has been isolated from a HeLa cell cDNA library. Analysis of the plasmid clone indicated that the insert has an ORF identical to that of hIMPase (human myo-inositol monophosphatase). The ability of hIMPase to suppress galactose toxicity is sensitive to the presence of Li(+) in the medium. A gal7 yeast strain harboring a plasmid containing cloned hIMPase grows on galactose as a sole carbon source. hIMPase mediated galactose metabolism is dependent on the functionality of GAL1 as well as GAL10 encoded galactokinase and epimerase respectively. These results predicted that the UDP-glucose/galactose pyrophosphorylase mediated pathway may be responsible for the relief of galactose toxicity. Experiments conducted to test this prediction revealed that expression of UGP1 encoded UDP-glucose pyrophosphorylase can indeed overcome the relief of galactose toxicity. Moreover, expression of UGP1 allows a gal7 strain to grow on galactose as a sole carbon source. Unlike the hIMPase mediated relief of galactose toxicity, UGP1 mediated relief of galactose toxicity is lithium insensitive. Based on our results and on the basis of available information on galactose toxicity, we suggest an alternative explanation for the molecular mechanism of galactose toxicity. PMID- 10452957 TI - In vivo assessment of microcystin-LR-induced hepatotoxicity in the rat using proton nuclear magnetic resonance (1H-NMR) imaging. AB - Microcystin-LR (MCLR)-induced hepatotoxicity was assessed in vivo in male Sprague Dawley rats (150-350 g) using magnetic resonance imaging (MRI). Following the intraperitoneal administration of MCLR (LD(50)), a region of damage, characterised by increased signal intensity on T(2)-weighted images, was seen proximal to the hepatic portal vein in the liver. Similarly, increased signal intensity was seen in the chemical-shift selective images (CSSI) of water frequency, proximal to the hepatic portal vein in the liver. This indicates that the increased signal intensity observed in the T(2)-weighted images was due to an increased amount of magnetic resonance (MR) visible protons in the tissue which represents an oedematous response. Image analysis of regions of apparent damage around the hepatic portal vein indicated a statistically significant increase in signal intensity in this region. Mitochondrial swelling and lipid inclusions were observed by transmission electron microscopy (TEM) in samples obtained from the oedematous regions of the liver using spatial coordinates from the magnetic resonance (MR) images. Massive haemorrhagic necrosis and nuclear swelling were observed by light microscopy in the centrilobular regions of the lobules. PMID- 10452958 TI - Characterization and purification of an outer membrane metalloproteinase from Pseudomonas aeruginosa with fibrinogenolytic activity. AB - A membrane proteinase from Pseudomonas aeruginosa, called insulin-cleaving membrane proteinase (ICMP), was located in the outer membrane leaflet of the cell envelope. The enzyme is expressed early in the logarithmic phase parallel to the bacterial growth during growth on peptide rich media. It is located with its active center facing to the outermost side of the cell, because its whole activity could be measured in intact cells. The very labile membrane proteinase was solubilized by non-ionic detergents (Nonidet P-40, Triton X-100) and purified in its amphiphilic form to apparent homogeneity in SDS-PAGE by copper chelate chromatography and two subsequent chromatographic steps on Red-Sepharose CL-4B (yield 58.3%, purification factor 776.3). It consisted of a single polypeptide chain with a molecular mass of 44.6 kDa, determined by mass spectrometry. ICMP was characterized to be a metalloprotease with pH-optimum in the neutral range. The ICMP readily hydrolyzed Glu(13)-Ala(14) and Tyr(16)-Leu(17) bonds in the insulin B-chain. Phe(25)-Tyr(26) and His(10)-Leu(11) were secondary cleavage sites suggesting a primary specificity of the enzyme for hydrophobic or aromatic residues at P'(1)-position. The ICMP differed from elastase, alkaline protease and LasA in its cleavage specificity, inhibition behavior and was immunologically diverse from elastase. The amino acid sequence of internal peptides showed no homologies with the known proteinases. This outer membrane proteinase was capable of specific cleavage of alpha and beta fibrinogen chains. Among the p nitroanilide substrates tested, substrates of plasminogen activator, complement convertase and kallikrein with arginine residues in the P(1)-subsite were the substrates best accepted, but they were only cleaved at a very low rate. PMID- 10452959 TI - Obese mice have higher insulin receptor levels in the hepatocyte cell nucleus following insulin stimulation in vivo with an oral glucose meal. AB - Internalization of the insulin receptor occurs following insulin binding at the cell surface, which serves to attenuate the insulin signal as well as modulate the number of surface insulin receptors. Obese animals exhibit decreased cell surface insulin receptor number as well as defects in insulin receptor internalization and processing. The insulin receptor may also translocates to the nucleus of hepatocytes and adipocytes following stimulation of cells with insulin. The objective of this study was to determine if insulin receptor trafficking to the hepatocyte cell nucleus could be observed in vivo and whether this process was altered in obese compared to lean mice. Mice were fasted for 12 h to reduce serum insulin to basal levels. Animals were then given an oral meal of glucose to stimulate the binding of insulin to receptor in vivo. Hepatocyte plasma membrane and nuclei were fractionated to purity following the glucose meal. Levels of insulin receptor were determined using insulin binding assays and a Western blotting assay using anti-insulin receptor antibody. As the amount of serum insulin increased following the glucose meal, a corresponding increase in nuclear insulin binding occurred in lean animals but not obese animals (P<0.05). Following the glucose meal, insulin receptor detected in the cell nucleus was increased in obese compared to lean mice (P<0.05). Thus insulin receptor translocation to the nucleus was demonstrated in vivo following a glucose meal in hepatocytes of both lean and obese animals. It is suggested that serum hyperglycemia and hyperinsulinemia in obese mice increased translocation of the insulin receptor to the nucleus. PMID- 10452960 TI - Aerosolized endotoxin is immediately bound by pulmonary surfactant protein D in vivo. AB - Collectins are carbohydrate binding proteins that are implicated in innate host defense. The lung collectins, surfactant proteins A and D (SP-A and SP-D), bind a variety of pathogens in vitro and influence phagocytosis by alveolar macrophages. In this report we show that SP-D binds endotoxin (lipopolysaccharide, LPS) in vivo in a rat model of acute respiratory distress syndrome (ARDS). Intratracheal aerosolization of LPS in rats resulted in the typical features of human ARDS. Total amounts of SP-D, as well as the carbohydrate binding properties of SP-D were measured in lung lavage as a function of time. The amount of SP-D did not change during 24 h. Interestingly, SP-D in lung lavage isolated from rats during the first 2 h after LPS treatment, was not able to bind to carbohydrate. Further analysis revealed that the carbohydrate binding sites of SP-D were occupied by LPS, suggesting that SP-D is an LPS scavenging molecule in vivo. Electron microscopic analysis indicated that, 1 h after LPS aerosolization, aggregates of SP-D with LPS were found in lysosomal structures in alveolar macrophages. We conclude that the lung collectin SP-D binds inhaled endotoxin in vivo, which may help to protect the lung from endotoxin-induced disease. PMID- 10452961 TI - A novel mutation in the CYBB gene resulting in an unexpected pattern of exon skipping and chronic granulomatous disease. AB - Chronic granulomatous disease is a rare inherited disorder caused by non-existent or severely decreased phagocyte superoxide production that results in a severe defect in host defense and consequent predisposition to microbial infection. The enzyme responsible for superoxide production, NADPH oxidase, involves at least five components. An absence of, or a defect in, any one of four of these proteins (p47(phox), p67(phox), p22(phox) and gp91(phox)) gives rise to the known types of chronic granulomatous disease. The most common form of inheritance is X-linked and is due to mutations in the CYBB gene that encodes gp91(phox), the large subunit of flavocytochrome b, the terminal electron donor of the oxidase. We have recently reported a large number of mutations in this gene revealing a broad range of defects, including large and small deletions, and frameshift, nonsense, missense, splice region and regulatory region mutations. Here we report a patient who has an unusual type of mutation that results in the generation of a 'pseudo exon' in the gp91(phox) mRNA and an unexpected pattern of splicing. PMID- 10452962 TI - Superoxide dismutase and hydrogen peroxide cause rapid nitric oxide breakdown, peroxynitrite production and subsequent cell death. AB - Isolated copper/zinc superoxide dismutase (Cu/Zn-SOD) or manganese superoxide dismutase (Mn-SOD) together with hydrogen peroxide (H(2)O(2)) caused rapid breakdown of nitric oxide (NO) and production of peroxynitrite (ONOO(-)) indicated by the oxidation of dihydrorhodamine-1,2,3 (DHR) to rhodamine-1,2,3. The breakdown of NO by this reaction was inhibited by cyanide (CN(-)) or by diethyldithiocarbamate (DETC), both Cu/Zn-SOD inhibitors, and the conversion of DHR to rhodamine-1,2,3 was inhibited by incubating Cu/Zn-SOD with either CN(-) or with high levels of H(2)O(2) or by including urate, a potent scavenger of ONOO( ). In the presence of phenol, the reaction of SOD, H(2)O(2) and NO caused nitration of phenol, which is known to be a footprint of ONOO(-) formation. H(2)O(2) addition to macrophages (cell line J774) expressing the inducible form of NO synthase (i-NOS) caused rapid breakdown of the NO they produced and this was also inhibited by CN(-) and by DETC. Subsequent ONOO(-) production by the macrophages, via this reaction, was inhibited by CN(-), high levels of H(2)O(2) or by urate. H(2)O(2) addition to i-NOS macrophages also caused cell death which was, in part, prevented by DETC or urate. We also found inhibition of mitochondrial respiration with malate and pyruvate as substrates, when isolated liver mitochondria were incubated with Cu/Zn-SOD, H(2)O(2) and NO. Inhibition of mitochondrial respiration was partly prevented by urate. The production of ONOO( ) by SOD may be of significant importance pathologically under conditions of elevated H(2)O(2) and NO levels, and might contribute to cell death in inflammatory and neurodegenerative diseases, as well as in macrophage-mediated host defence. PMID- 10452963 TI - Is gut the major source of proinflammatory cytokine release during polymicrobial sepsis? AB - Although studies have shown that the gut is capable of being a cytokine-producing organ and that the proinflammatory cytokines TNF-alpha, IL-1beta, and IL-6 are upregulated following the onset of sepsis, it remains unknown whether the gut is indeed the major source of the increased cytokine production under such conditions. To determine this, male rats were subjected to cecal ligation and puncture (CLP, a model of polymicrobial sepsis) or sham operation followed by the administration of normal saline solution subcutaneously (i.e., fluid resuscitation). Systemic and portal blood samples were taken simultaneously at 2, 5, 10, or 20 h after CLP or sham operation. Plasma levels of TNF-alpha, IL-1beta, and IL-6 were determined using an enzyme-linked immunosorbent assay. In additional animals, the small intestine was harvested at 10 h after CLP or sham operation and examined for TNF-alpha, IL-1beta, and IL-6 gene expression by RT PCR. The results indicate that the levels of TNF-alpha, IL-1beta, and IL-6 in both systemic and portal blood samples were significantly elevated during sepsis with the exception that the increase in IL-1beta was not significant at 2 h after CLP. However, there were no significant differences in the levels of those proinflammatory cytokines between systemic and portal blood at any points after the onset of sepsis. Moreover, there were no significant alterations in the proinflammatory cytokine gene expression in the small intestine at 10 h after CLP. Since the levels of TNF-alpha, IL-1beta, and IL-6 were not significantly increased in portal blood as compared to systemic blood and since there was no upregulation of gene expression for these cytokines, it appears that organs other than the gut are responsible for the upregulated proinflammatory cytokines during polymicrobial sepsis. PMID- 10452964 TI - Glycation of apolipoprotein E impairs its binding to heparin: identification of the major glycation site. AB - The increased glycation of plasma apolipoproteins represents a possible major factor for lipid disturbances and accelerated atherogenesis in diabetic patients. The glycation of apolipoprotein E (apoE), a key lipid-transport protein in plasma, was studied both in vivo and in vitro. ApoE was shown to be glycated in plasma very low density lipoproteins of both normal subjects and hyperglycemic, diabetic patients. However, diabetic patients with hyperglycemia showed a 2-3 fold increased level of apoE glycation. ApoE from diabetic plasma showed decreased binding to heparin compared to normal plasma apoE. The rate of Amadori product formation in apoE in vitro was similar to that for albumin and apolipoproteins A-I and A-II. The glycation of apoE in vitro significantly decreased its ability to bind to heparin, a critical process in the sequestration and uptake of apoE-containing lipoproteins by cells. Diethylenetriaminepentaacetic acid, a transition metal chelator, had no effect on the loss of apoE heparin-binding activity, suggesting that glycation rather than glycoxidation is responsible for this effect. In contrast, glycation had no effect on the interaction of apoE with amyloid beta-peptide. ApoE glycation was demonstrated to be isoform-specific. ApoE(2) showed a higher glycation rate and the following order was observed: apoE(2)>apoE(4)>apoE(3). The major glycated site of apoE was found to be Lys-75. These findings suggest that apoE is glycated in an isoform-specific manner and that the glycation, in turn, significantly decreases apoE heparin-binding activity. We propose that apoE glycation impairs lipoprotein-cell interactions, which are mediated via heparan sulfate proteoglycans and may result in the enhancement of lipid abnormalities in hyperglycemic, diabetic patients. PMID- 10452965 TI - Cutting edge: developmental switches in chemokine responses during T cell maturation. AB - We show that developmental transitions during thymocyte maturation are associated with dramatic changes in chemotactic responses to chemokines. Macrophage-derived chemokine, a chemokine expressed in the thymic medulla, attracts thymocytes only during a brief window of development, between the late cortical and early medullary stages. All medullary phenotypes (CD4 or CD8 single positive) but not immature thymocytes respond to the medullary stroma-expressed (and secondary lymphoid tissue-associated) chemokines secondary lymphoid-tissue chemokine and macrophage inflammatory protein-3beta. The appearance of these responses is associated with the phenotypic stage of cortex to medulla migration and with up regulation of mRNA for the receptors CCR4 (for macrophage-derived chemokine and thymus and activation-regulated chemokine) and CCR7 (for secondary lymphoid tissue chemokine and macrophage inflammatory protein-3beta). In contrast, most immature and medullary thymocytes migrate to thymus-expressed chemokine, an ability that is lost only with up-regulation of the peripheral homing receptor L selectin during the latest stages of thymocyte maturation associated with export to the periphery. Developmental switches in chemokine responses may help regulate critical migratory events during T cell development. PMID- 10452967 TI - Cutting edge: N-hydroxy peptides: a new class of TCR antagonists. AB - TCR antagonists are altered T cell epitopes that specifically inactivate T cells. Commonly, they are derived from agonists by amino acid side chain replacement at positions accessible to the TCR. In this paper we report for the first time that a main chain N-hydroxylation, which is not exposed at the surface of the MHC peptide complex, renders an agonist into an antagonist. These mimotopes are a new, yet undescribed class of TCR antagonists. The antagonists are about 100 times more potent than an unrelated peptide that competes for binding to the MHC molecule. The novel main chain modification enhances biostability and maintains side chain constitution and thus opens new prospects for the use of TCR antagonists in the treatment of pathological immune reactions. PMID- 10452966 TI - Cutting edge: blockade of the CD28/B7 costimulatory pathway inhibits intestinal allograft rejection mediated by CD4+ but not CD8+ T cells. AB - The effect of blocking the CD28/B7 costimulatory pathway on intestinal allograft rejection was examined in mice. Murine CTLA4Ig failed to prevent the rejection of allografts transplanted into wild-type or CD4 knockout (KO) mice but did inhibit allograft rejection by CD8 KO recipients. This effect was associated with decreased intragraft mRNA for IFN-gamma and TNF-alpha and increased mRNA for IL-4 and IL-5. This altered pattern of cytokine production was not observed in allografts from murine CTLA4Ig-treated CD4 KO mice. These data demonstrate that blockade of the CD28/B7 pathway has different effects on intestinal allograft rejection mediated by CD4+ and CD8+ T cells and suggest that these T cell subsets have different costimulatory requirements in vivo. The results also suggest that the inhibition of CD4+ T cell-mediated allograft rejection by CTLA4Ig may be related to down-regulation of Th1 cytokines and/or up-regulation of Th2 cytokines. PMID- 10452968 TI - Cutting edge: CXCR4-Lo: molecular cloning and functional expression of a novel human CXCR4 splice variant. AB - Human CXCR4 is a specific receptor for the CXC chemokine stromal cell-derived factor-1 (SDF-1) and a coreceptor for T cell line tropic strains of HIV-1. Genetic knockouts of CXCR4 and SDF-1 have delineated their critical role during embryonic cardiogenesis, leukopoiesis, and vasculogenesis. Herein, we used bioinformatics and differential strategies like isoform-specific RT-PCR and Northern blots to identify and clone a novel unspliced isoform of human CXCR4, termed CXCR4-Lo. CXCR4-Lo corresponds to a larger approximately 4. 0-kb mRNA transcript and differs from the known human CXCR4 by the first 9 aa in the functionally important NH2-terminal extracellular domain of the receptor. CXCR4 Lo-transfected rat basophil leukemia-2H3 cells responded to SDF-1 with a transient rise of intracellular Ca2+ concentration and by undergoing chemotaxis. Expression of CXCR4-Lo is noteworthy, as it may have differential affinity as a coreceptor for HIV strains in comparison with CXCR4. Furthermore, CXCR4-Lo may also provide a functional backup to CXCR4 during embryogenesis. PMID- 10452969 TI - Cutting edge: activation of NK T cells by CD1d and alpha-galactosylceramide directs conventional T cells to the acquisition of a Th2 phenotype. AB - NK T cells recognize glycolipid Ags such as alpha-galactosylceramide (alpha GalCer) presented by the MHC class I-like molecule CD1d. In this paper we have studied the in vivo effects of alpha-GalCer on the generation of adaptive immune responses. Treatment of mice with alpha-GalCer resulted in rapid activation of NK T cells and production of the cytokines IL-4 and IFN-gamma. However, after this initial stimulation, NK T cells became polarized for the production of IL-4. Further, as soon as 6 days after alpha-GalCer injection, a marked increase in serum IgE levels was observed. Administration of alpha-GalCer at the time of priming of mice with protein Ag resulted in the generation of Ag-specific Th2 cells and a profound increase in the production of IgE. Collectively, these findings indicate that alpha-GalCer may be useful for modulating immune responses toward a Th2 phenotype during prophylaxis and therapy. PMID- 10452970 TI - Cutting edge: a novel mechanism for rescue of B cells from CD95/Fas-mediated apoptosis. AB - CD95-induced apoptosis contributes to the maintenance of homeostasis in both B and T lymphocyte-mediated immunity. B cells increase CD95 expression in response to activation signals and become susceptible to CD95-induced apoptosis. Protection from CD95-mediated death signals can be induced in mature B cells by signals delivered through the B cell Ag receptor. In this paper we demonstrate for the first time that rescue from apoptosis can occur independently of de novo protein synthesis. This rescue from apoptosis prevents activation of caspase 8, the apical caspase in the CD95 death pathway, and CD95-FADD (Fas-associated death domain containing protein) association does not occur normally. Thus B cell activation signals can biochemically modify proximal elements of the CD95 death pathway and regulate the sensitivity of cells to apoptosis induction at an early stage in programmed cell death. PMID- 10452972 TI - Cutting edge: inhibition of experimental tumor metastasis by dendritic cells pulsed with alpha-galactosylceramide. AB - A unique lymphoid lineage, Valpha14 NKT cells, bearing an invariant Ag receptor encoded by Valpha14 and Jalpha281 gene segments, play crucial roles in various immune responses, including protective immunity against malignant tumors. A specific ligand of Valpha14 NKT cells is determined to be alpha galactosylceramide (alpha-GalCer) which is presented by the CD1d molecule. Here, we report that dendritic cells (DCs) pulsed with alpha-GalCer effectively induce potent antitumor cytotoxic activity by specific activation of Valpha14 NKT cells, resulting in the inhibition of tumor metastasis in vivo. Moreover, a complete inhibition of B16 melanoma metastasis in the liver was observed when alpha-GalCer pulsed DCs were injected even 7 days after transfer of tumor cells to syngeneic mice where small but multiple metastatic nodules were already formed. The potential utility of DCs pulsed with alpha-GalCer for tumor immunotherapy is discussed. PMID- 10452971 TI - Cutting edge: inflammatory signaling by Borrelia burgdorferi lipoproteins is mediated by toll-like receptor 2. AB - The agent of Lyme disease, Borrelia burgdorferi, produces membrane lipoproteins possessing potent inflammatory properties linked to disease pathology. The recent association of toll-like receptors (TLR) 2 and 4 with LPS responses prompted the examination of TLR involvement in lipoprotein signaling. The ability of human cell lines to respond to lipoproteins was correlated with the expression of TLR2. Transfection of TLR2 into cell lines conferred responsiveness to lipoproteins, lipopeptides, and sonicated B. burgdorferi, as measured by nuclear translocation of NF-kappaB and cytokine production. The physiological importance of this interaction was demonstrated by the 10-fold greater sensitivity of TLR2 transfected cells to lipoproteins than LPS. Futhermore, TLR2-dependent signaling by lipoproteins was facilitated by CD14. These data indicate that TLR2 facilitates the inflammatory events associated with Lyme arthritis. In addition, the widespread expression of lipoproteins by other bacterial species suggests that this interaction may have broad implications in microbial inflammation and pathogenesis. PMID- 10452973 TI - Distribution fate and mechanism of immune modulation following mucosal delivery of plasmid DNA encoding IL-10. AB - DNA vaccination has been widely studied in several models of vaccination and in the treatment of inflammatory diseases, even though the mechanism involved is still unclear. This report demonstrates that mucosal administration of plasmid DNA leads to rapid and widespread distribution around the body. Dissemination likely occurred via the bloodstream because plasmid DNA was present in blood plasma. The plasmid DNA was also detectable in several tissues including draining lymph nodes, spleen, liver, bone marrow, and even the dermis of ear pinnae. Except for the site of administration, plasmid DNA was no longer detectable in tissues after 3 wk postadministration. RNA and protein expression was also found in the tissues and bloodstream. Animals previously primed by HSV infection and subsequently given IL-10 DNA via the nasal mucosa, showed diminished Ag-induced delayed type hypersensitivity reactions for up to 5 wk posttreatment. The mechanism of modulation involved diminished the Ag-specific proliferation and production of Th1 cytokines. The Ag-specific silencing effects persisted beyond the duration of detectable plasmid encoded protein and was maintained upon adoptive transfer of T cells into a plasmid-free environment. The silenced T cells were not a source of IL-10, and their anergic state was reversible by exposure to Ag in the presence of exogenous IL-2. PMID- 10452974 TI - Caspase-1 regulates the inflammatory process leading to autoimmune demyelination. AB - T cell-mediated inflammation is considered to play a key role in the pathogenic mechanisms sustaining multiple sclerosis (MS). Caspase-1, formerly designated IL 1beta-converting enzyme, is crucially involved in immune-mediated inflammation because of its pivotal role in regulating the cellular export of IL-1beta and IL 18. We studied the role of caspase-1 in experimental autoimmune encephalomyelitis (EAE), the animal model for MS. Caspase-1 is transcriptionally induced during EAE, and its levels correlate with the clinical course and transcription rate of proinflammatory cytokines such as TNF-alpha, IL-1beta, IFN-gamma, and IL-6. A reduction of EAE incidence and severity is observed in caspase-1-deficient mice, depending on the immunogenicity and on the amount of the encephalitogenic myelin oligodendrocyte glycoprotein (MOG) peptide used. In caspase-1-deficient mice, reduced EAE incidence correlates with defective development of anti-MOG IFN-gamma producing Th1 cells. Finally, pharmacological blockade of caspase-1 in Biozzi AB/H mice, immunized with spinal cord homogenate or MOG35-55 peptide, by the caspase-1-inhibitor Z-Val-Ala-dl -Asp-fluoromethylketone, significantly reduces EAE incidence in a preventive but not in a therapeutic protocol. These results indicate that caspase-1 plays an important role in the early stage of the immune mediated inflammatory process leading to EAE, thus representing a possible therapeutic target in the acute phase of relapsing remitting MS. PMID- 10452975 TI - Direct neurite-mast cell communication in vitro occurs via the neuropeptide substance P. AB - Communication between nerves and mast cells is a prototypic demonstration of neuroimmune interaction. However, whether mast cell activation occurs as a direct response to neuronal activation or requires an intermediary cell is unclear. Addressing this issue, we used an in vitro coculture approach comprising cultured murine superior cervical ganglia and rat leukemia basophilic cells (RBLs; possesses properties of mucosal-type mast cells). Following loading with the calcium fluorophore, Fluo-3, neurite-RBL units (separated by <50 nm) were examined by confocal laser scanning microscopy. Addition of bradykinin, or scorpion venom, dose-dependently elicited neurite activation (i.e., Ca2+ mobilization) and, after a lag period, RBL Ca2+ mobilization. Neither bradykinin nor scorpion venom had any direct effect on the RBLs in the absence of neurites. Addition of a neutralizing substance P Ab or a neurokinin (NK)-1 receptor antagonist, but not an NK-2 receptor antagonist, dose-dependently prevented the RBL activation that resulted as a consequence of neural activation by either bradykinin or scorpion venom. These data illustrate that nerve-mast cell cross talk can occur in the absence of an intermediary transducing cell and that the neuropeptide substance P, operating via NK-1 receptors, is an important mediator of this communication. Our findings have implications for the neuroimmune signaling cascades that are likely to occur during airways inflammation, intestinal hypersensitivity, and other conditions in which mast cells feature. PMID- 10452976 TI - Inefficient phospholipase C activation and reduced Lck expression characterize the signaling defect of umbilical cord T lymphocytes. AB - Adult and neonatal immunocompetent cells exhibit important functional distinctions, including differences in cytokine production and susceptibility to tolerance induction. We have investigated the molecular features that characterize the immune response of cord blood-derived T lymphocytes compared with that of adult T lymphocytes. Our findings demonstrate that phospholipase C (PLC) isozymes, which play a pivotal role in the control of protein kinase C activation and Ca2+ mobilization, are differently expressed in cord and adult T lymphocytes. PLCbeta1 and delta1 are expressed at higher levels in cord T cells, while PLCbeta2 and gamma1 expression is higher in adult T lymphocytes. PLCdelta2 and gamma2 appear to be equally expressed in both cell types. In addition, a functional defect in PLC activation via CD3 ligation or pervanadate treatment, stimuli that activate tyrosine kinases, was observed in cord blood T cells, whereas treatment with aluminum tetrafluoride (AlF4-), a G protein activator, demonstrated a similar degree of PLC activation in cord and adult T cells. The impaired PLC activation of cord blood-derived T cells was associated with a a very low expression of the Src kinase, Lck, along with a reduced level of ZAP70. No mitogenic response to CD3 ligation was observed in cord T cells. However, no signaling defect was apparent downstream of PLC activation, as demonstrated by the mitogenic response of cord T cells to the pharmacologic activation of protein kinase C and Ca2+ by treatment with PMA and ionomycin. Thus, neonatal cord blood derived T cells show a signaling immaturity associated with inadequate PLCgamma activation and decreased Lck expression. PMID- 10452977 TI - Reduced IL-4-, lipopolysaccharide-, and IFN-gamma-induced MHC class II expression in mice lacking class II transactivator due to targeted deletion of the GTP binding domain. AB - Class II transactivator (CIITA) is an unusual transcriptional coactivator in that it contains a functionally important, GTP-binding consensus domain. To assess the functional role of the GTP-binding domain of CIITA in vivo, we have generated knockout mice that bear a mutation in the CIITA gene spanning the GTP-binding domain. Upon analysis, these mice show no detectable CIITA mRNA; hence, they represent mice with deleted CIITA rather than mice with defects in the GTP binding domain only. In these knockout mice, MHC class II expression is nearly eliminated, although a faint RT-PCR signal is visible in spleen, lymph node, and thymus, suggestive of the presence of CIITA-independent regulation of MHC class II expression. Invariant chain expression is also greatly reduced, but to a lesser extent than MHC class II. Serum IgM is not decreased, but the serum IgG level is greatly reduced, further confirming the absence of MHC class II Ag dependent Ig class switching. Induction of MHC class II expression by IL-4 or LPS was absent on B cells, and Mac-1+ cells showed no detectable induction of MHC class II by either IL-4, LPS, or IFN-gamma. These findings demonstrate a requirement for CIITA in IFN-gamma-, IL-4-, and endotoxin-induced MHC class II expression as well as the possibility of rare CIITA-independent MHC class II expression. PMID- 10452978 TI - CD28 costimulation accelerates IL-4 receptor sensitivity and IL-4-mediated Th2 differentiation. AB - The development of Th1 and Th2 cells is determined by the type of antigenic stimulation involved in the initial cell activation step. Evidence indicates that costimulatory signals, such as those delivered by CD28, play an important role in Th2 development, but little is known about how CD28 costimulation contributes to Th2 development. In this study, TCR cross-linking was insufficient for Th2 development, while the addition of CD28 costimulation drastically increased Th2 generation through the IL-4-mediated pathway. Th2 generation following CD28 costimulation was not simply explained by the enhancement of IL-4 production in naive T cells. To generate Th2 cells after TCR cross-linking only, it was necessary to add a 20- to 200-fold excess of IL-4 generated after TCR and CD28 stimulation. TCR cross-linking increased the expression level and binding property of the IL-4R, but enhanced the sensitivity to IL-4 only slightly. In contrast, as evidenced by the enhanced phosphorylation of Jak3, the IL-4Ralpha chain, and STAT6 following IL-4 stimulation, CD28 costimulation increased IL-4R sensitivity without affecting its expression and binding property. This evidence of the enhancement of IL-4R sensitivity increases our understanding of how CD28 costimulation accelerates Th2 development. PMID- 10452979 TI - Cytokine deprivation of naive CD8+ T cells promotes minimal cell cycling but maximal cytokine synthesis and autonomous proliferation subsequently: a mechanism of self-regulation. AB - Naive CD8+ T cells differentiate into effectors secreting various cytokines that aid their function. IL-2, but not IL-15, promoted this differentiation of naive CD8+ T cells into effectors. However, the amount of IL-2 present during differentiation had a dichotomous effect on their subsequent function. High concentrations of IL-2 enhanced proliferation and cell cycling initially, but the effectors subsequently failed to produce cytokines and proliferate autonomously, although CD28 expression was maintained. In contrast, suboptimal amounts of IL-2 during priming promoted apoptosis, little proliferation and cell cycling, yet the CD8+ effectors generated produced high levels of cytokines and proliferated autonomously. Interestingly, the effects of IL-2 on naive CD8+ T cells were totally opposite those on naive CD4+ T cells. Although IL-2 impaired cytokine synthesis by CD8+ T cells, the expression of LFA1 and CD44 as well as Fas dependent cytotoxicity were enhanced. However, loss of cytokine synthesis was not due to increased cytotoxicity, as inhibition occurred even in the absence of perforin/FasL. Interestingly, CD8+ effectors secreting reduced amounts of cytokines exhibited enhanced IL-2Ralpha, but reduced IL-2Rbeta, expression. Furthermore, sorted CD8+ IL-2Ralphahigh cells secreted less cytokines than IL 2Ralphalow cells. These results suggest that the presence of excessive IL-2 during the activation of naive CD8+ T cells, while promoting cell cycling initially, may compromise long-term immunity. PMID- 10452980 TI - Macrophage colony-stimulating factor induces the expression of mitogen-activated protein kinase phosphatase-1 through a protein kinase C-dependent pathway. AB - M-CSF triggers the activation of extracellular signal-regulated protein kinases (ERK)-1/2. We show that inhibition of this pathway leads to the arrest of bone marrow macrophages at the G0/G1 phase of the cell cycle without inducing apoptosis. M-CSF induces the transient expression of mitogen-activated protein kinase phosphatase-1 (MKP-1), which correlates with the inactivation of ERK-1/2. Because the time course of ERK activation must be finely controlled to induce cell proliferation, we studied the mechanisms involved in the induction of MKP-1 by M-CSF. Activation of ERK-1/2 is not required for this event. Therefore, M-CSF activates ERK-1/2 and induces MKP-1 expression through different pathways. The use of two protein kinase C (PKC) inhibitors (GF109203X and calphostin C) revealed that M-CSF induces MKP-1 expression through a PKC-dependent pathway. We analyzed the expression of different PKC isoforms in bone marrow macrophages, and we only detected PKCbetaI, PKCepsilon, and PKCzeta. PKCzeta is not inhibited by GF109203X/calphostin C. Of the other two isoforms, PKCepsilon is the best candidate to mediate MKP-1 induction. Prolonged exposure to PMA slightly inhibits MKP-1 expression in response to M-CSF. In bone marrow macrophages, this treatment leads to a complete depletion of PKCbetaI, but only a partial down-regulation of PKCepsilon. Moreover, no translocation of PKCbetaI or PKCzeta from the cytosol to particulate fractions was detected in response to M-CSF, whereas PKCepsilon was constitutively present at the membrane and underwent significant activation in M CSF-stimulated macrophages. In conclusion, we remark the role of PKC, probably isoform epsilon, in the negative control of ERK-1/2 through the induction of their specific phosphatase. PMID- 10452981 TI - A mathematical model on germinal center kinetics and termination. AB - We devise a mathematical model to study germinal center (GC) kinetics. Earlier models for GC kinetics are extended by explicitly modeling 1) the cell division history of centroblasts, 2) the Ag uptake by centrocytes, and 3) T cell dynamics. Allowing for T cell kinetics and T-B cell interactions, we study the role of GC T cells in GC kinetics, GC termination, and B cell selection. We find that GC T cells play a major role in GC formation, but that the maintenance of established GC reactions requires very few T cells only. The results therefore suggest that the termination of a GC reaction is largely caused by lack of Ag on the follicular dendritic cells and is hardly influenced by Th cells. Ag consumption by centrocytes is the major factor determining the decay rate of the antigenic stimulus during a GC reaction. Investigating the effect of the Ag dose on GC kinetics, we find that both the total size of the GC and its duration are hardly influenced by the initial amount of Ag. In the model this is due to a buffering effect by competition for limited T cell help and/or competition between proliferating centroblasts. PMID- 10452982 TI - Disruption of CD154:CD40 blocks generation of allograft immunity without affecting APC activation. AB - CD154 (CD40 ligand, gp39) interaction with its receptor CD40 has been shown to be critically important for the generation of cell-mediated as well as humoral immunity. It has been proposed that ligation of CD40 on APCs, presumably by activated Th cells, leads to increased APC function as defined by up-regulation of costimulatory molecules and enhancement of IL-12 production. In this report, we directly examined the contribution of the CD154:CD40 pathway in a murine model of allograft rejection. Generation of both the CTL and alloantibody responses following injection with allogeneic P815 tumor cells was severely compromised in CD154 knockout mice and wild-type C57BL/6 mice treated with the anti-CD154 mAb, MR1. Splenic production of IL-2, IFN-gamma, and TNF was significantly suppressed from CD154-deficient mice, indicating a lack of T cell priming. However, splenic cells from CD154 knockout mice induced comparable levels of CD86 expression and IL-12 production when compared with their wild-type littermates. The treatment of CD154-/- mice with the agonistic anti-CD40 mAb, FGK45, generated activated APCs yet failed to restore either the CTL or alloantibody responses to P815. Likewise, immunization with B7-transfected P815 tumor cells failed to generate expansion of the CTL effector population in CD154-/- mice. These results suggest that the generation of allograft immunity is dependent on the interaction of CD154 with CD40 but not primarily for the activation of APCs. PMID- 10452983 TI - Germinal center B cell apoptosis requires both caspase and cathepsin activity. AB - Follicular dendritic cells (FDCs) select B cells during germinal center (GC) reactions. The B cells that are able to bind to the FDCs receive a signal that leads to the termination of endonuclease activity in the nuclei of those B cells. This signal must be in addition to the signals transferred through the cross linkage of the B cell receptors and signals resulting from the interactions of the adhesion molecules lymphocyte function-associated Ag-1 and very late Ag-4 with ICAM-1 and VCAM-1, respectively. In this report, we present evidence that the FDCs silence all apoptotic processes in GC B lymphocytes and additionally switch off pre-present endonuclease activity. We also show that GC B cell apoptosis requires cathepsin activity downstream of caspase-3. This cathepsin activity is directly connected to endonuclease activity and therefore may be an interesting target for the antiapoptotic factors produced by FDCs. PMID- 10452984 TI - B cell apoptosis triggered by antigen receptor ligation proceeds via a novel caspase-dependent pathway. AB - In contrast to positive signaling leading to proliferation, the mechanisms involved in negative signaling culminating in apoptosis after B cell Ag receptor (BCR) ligation have received little study. We find that apoptosis induced by BCR cross-linking on EBV-negative mature and immature human B cell lines involves the following sequential, required events: a cyclosporin A-inhibitable, likely calcineurin-mediated step; and activation of caspase-2, -3, and -9. Caspase-2 is activated early and plays a major role in the apoptotic pathway, while caspase-9 is activated later in the apoptotic pathway and most likely functions to amplify the apoptotic signal. Caspase-8 and -1, which are activated by ligation of the CD95 and TNF-R1 death receptors, are not involved. Apoptosis induced by BCR ligation thus proceeds via a previously unreported intracellular signaling pathway. PMID- 10452985 TI - Norepinephrine-mediated inhibition of antitumor cytotoxic T lymphocyte generation involves a beta-adrenergic receptor mechanism and decreased TNF-alpha gene expression. AB - We have previously shown that norepinephrine (NE) inhibits the in vitro generation of anti-MOPC-315 CTL activity by spleen cells from BALB/c mice rejecting a large MOPC-315 tumor as a consequence of low-dose melphalan (l phenylalanine mustard (l -PAM)) treatment (l -PAM TuB spleen cells). Since TNF alpha plays a key role in the generation of antitumor CTL activity in this system, we determined whether NE mediates this inhibition through inhibition of TNF-alpha production. Here, we show that NE inhibits the production of TNF-alpha protein and mRNA by l -PAM TuB spleen cells stimulated in vitro with mitomycin C treated tumor cells. Flow cytometric analysis of intracellular expression of TNF alpha revealed substantial NE-mediated decreases in the percentages of TNF-alpha+ cells among CD4+ and CD8+ T cells and F4/80+ activated macrophages. NE inhibition of CTL generation was largely overcome by addition of TNF-alpha to the stimulation cultures. When the beta-adrenergic antagonist propranolol was added to the stimulation cultures of l -PAM TuB spleen cells at a concentration that prevented NE-induced cAMP elevation, the NE-mediated decrease in TNF-alpha mRNA and NE-mediated inhibition of CTL generation were reversed. Collectively, these results suggest that NE inhibits antitumor CTL generation, at least in part, by inhibiting TNF-alpha synthesis through a mechanism(s) involving beta-adrenergic receptor signaling. PMID- 10452986 TI - Induction of allograft tolerance in the absence of Fas-mediated apoptosis. AB - Using certain immunosuppressive regimens, IL-2 knockout (KO) mice, in contrast to wild-type (wt) controls, are resistant to the induction of allograft tolerance. The mechanism by which IL-2 regulates allograft tolerance is uncertain. As IL-2 KO mice have a profound defect in Fas-mediated apoptosis, we hypothesized that Fas-mediated apoptosis of alloreactive T cells may be critical in the acquisition of allograft tolerance. To definitively study the role of Fas in the induction of transplantation tolerance, we used Fas mutant B6.MRL-lpr mice as allograft recipients of islet and vascularized cardiac transplants. Alloantigen-stimulated proliferation and apoptosis of Fas-deficient cells were also studied in vivo. Fas mutant B6.MRL-lpr (H-2b) mice rapidly rejected fully MHC-mismatched DBA/2 (H-2d) islet allografts and vascularized cardiac allografts with a tempo that is comparable to wt control mice. Both wt and B6.MRL-lpr mice transplanted with fully MHC-mismatched islet allografts or cardiac allografts can be readily tolerized by either rapamycin or combined costimulation blockade (CTLA-4Ig plus anti-CD40L mAb). Despite the profound defect of Fas-mediated apoptosis, Fas deficient T cells can still undergo apoptotic cell death in vivo in response to alloantigen stimulation. Our study suggests that: 1) Fas is not necessarily essential for allograft tolerance, and 2) Fas-mediated apoptosis is not central to the IL-2-dependent mechanism governing the acquisition of allograft tolerance. PMID- 10452987 TI - CD45 is essential for Fc epsilon RI signaling by ZAP70, but not Syk, in Syk negative mast cells. AB - The ZAP70/Syk family of protein tyrosine kinases plays an important role in Ag receptor signaling. Structural similarity of Syk and ZAP70 suggests their functional overlap. Previously, it was observed that expression of either ZAP70 or Syk reconstitutes Ag receptor signaling in Syk-negative B cells. However, in CD45-deficient T cells, Syk, but not ZAP70, restores T cell receptor-signaling pathway. To study the function of Syk, ZAP70, and CD45 in mast cells, a Syk/CD45 double-deficient variant of RBL-2H3 cells was characterized. After transfection, stable cell lines were isolated that expressed ZAP70, Syk, CD45, ZAP70 plus CD45, and Syk plus CD45. IgE stimulation did not induce degranulation in parental double-deficient cells, nor in the cells expressing only CD45. ZAP70 expression did not restore Fc epsilon RI signaling unless CD45 was coexpressed in the cells. However, Syk alone restored the IgE signal transduction pathway. The coexpression of CD45 with Syk had no significant effects on the responses to FcepsilonRI aggregation. There was much better binding of Syk than ZAP70 to the phosphorylated Fc epsilon RI gamma-ITAM. Furthermore, unlike Syk, ZAP70 required CD45 to display receptor-induced increase in kinase activity. Therefore, in mast cells, ZAP70, but not Syk, requires CD45 for Ag receptor-induced signaling. PMID- 10452988 TI - Autocrine IL-12 is involved in dendritic cell modulation via CD40 ligation. AB - Ligation of CD40 on dendritic cells (DC) triggers production of IL-12. Using an adoptive transfer model we have previously shown that rIL-12 acts directly on DC to enhance presentation of an otherwise poorly immunogenic tumor peptide. Using the same experimental model, we now describe a similar adjuvanticity of CD40 ligation on peptide presentation by DC. We also explore the possibility that the IL-12 resulting from CD40 ligation directly affects the APC function of DC, mediating or contributing to the adjuvant effect of CD40 ligation. CD40 engagement in vitro and rIL-12 at concentrations in the range induced by CD40 ligation were equally effective in priming DC for presentation of the tumor peptide in vivo. Remarkably, the copresence in vitro of neutralizing Ab to IL-12, but not to TNF-alpha, IL-1beta, or IFN-gamma, ablated the enhancing effect of CD40 engagement on the APC function of DC. These data suggest a major role for autocrine IL-12 in DC modulation via CD40 ligation. PMID- 10452989 TI - Differential responses of invariant V alpha 24J alpha Q T cells and MHC class II restricted CD4+ T cells to dexamethasone. AB - NK T cells are a T cell subset in the human that express an invariant alpha-chain (V alpha 24invt T cells). Because of the well-described immunomodulation by glucocorticoids on activation-induced cell death (AICD), the effects of dexamethasone and anti-CD3 stimulation on V alpha 24invt T cell clones and CD4+ T cell clones were investigated. Dexamethasone significantly enhanced anti-CD3 mediated proliferation of V alpha 24invt T cells, whereas CD4+ T cells were inhibited. Addition of neutralizing IL-2 Ab partially abrogated dexamethasone induced potentiation of V alpha 24invt T cell proliferation, indicating a role for autocrine IL-2 production in corticosteroid-mediated proliferative augmentation. Dexamethasone treatment of anti-CD3-stimulated V alpha 24invt T cells did not synergize with anti-Fas blockade in enhancing proliferation or preventing AICD. The V alpha 24invt T cell response to dexamethasone was dependent on the TCR signal strength. In the presence of dexamethasone, lower doses of anti-CD3 inhibited proliferation of V alpha 24invt T cells and CD4+ T cells; at higher doses of anti-CD3, which caused inhibition of CD4+ T cells, the V alpha 24invt T cell clones proliferated and were rescued from AICD. These results demonstrate significant differences in TCR signal strength required between V alpha 24invt T cells and CD4+ cells, and suggest important immunomodulatory consequences for endogenous and exogenous corticosteroids in immune responses. PMID- 10452990 TI - Inhibition of Kit expression by IL-4 and IL-10 in murine mast cells: role of STAT6 and phosphatidylinositol 3'-kinase. AB - The c-kit protooncogene encodes a receptor tyrosine kinase that is known to play a critical role in hemopoiesis and is essential for mast cell growth, differentiation, and cytokine production. Studies have shown that the Th2 cytokine IL-4 can down-regulate Kit expression on human and murine mast cells, but the mechanism of this down-regulation has remained unresolved. Using mouse bone marrow-derived mast cells, we demonstrate that IL-4-mediated Kit down regulation requires STAT6 expression and phosphotidylinositide-3'-kinase activation. We also find that the Th2 cytokine IL-10 potently down-regulates Kit expression. IL-4 enhances IL-10-mediated inhibition in a manner that is STAT6 independent and phosphotidylinositide-3'-kinase dependent. Both IL-4- and IL-10 mediated Kit down-regulation were coupled with little or no change in c-kit mRNA levels, no significant change in Kit protein stability, but decreased total Kit protein expression. Inhibition of Kit expression by IL-4 and IL-10 resulted in a loss of Kit-mediated signaling, as evidenced by reduced IL-13 and TNF-alpha mRNA induction after stem cell factor stimulation. These data offer a role for STAT6 and phosphotidylinositide-3'-kinase in IL-4-mediated Kit down-regulation, coupled with the novel observation that IL-10 is a potent inhibitor of Kit expression and function. Regulating Kit expression and signaling may be essential to controlling mast cell-mediated inflammatory responses. PMID- 10452991 TI - Polarized transport of MHC class II molecules in Madin-Darby canine kidney cells is directed by a leucine-based signal in the cytoplasmic tail of the beta-chain. AB - MHC class II molecules are found on the basolateral plasma membrane domain of polarized epithelial cells, where they can present Ag to intraepithelial lymphocytes in the vascular space. We have analyzed the sorting information required for efficient intracellular localization and polarized distribution of MHC class II molecules in stably transfected Madin-Darby canine kidney cells. These cells were able to present influenza virus particles to HLA-DR1-restricted T cell clones. Wild-type MHC class II molecules were located on the basolateral plasma membrane domain, in basolateral early endosomes, and in late multivesicular endosomes, the latter also containing the MHC class II-associated invariant chain and an HLA-DM fusion protein. A phenylalanine-leucine residue within the cytoplasmic tail of the beta-chain was required for basolateral distribution, efficient internalization, and localization of the MHC class II molecules to basolateral early endosomes. However, distribution to apically located, late multivesicular endosomes did not depend on signals in the class II cytoplasmic tails as both wild-type class II molecules and mutant molecules lacking the phenylalanine-leucine motif were found in these compartments. Our results demonstrate that sorting information in the tails of class II dimers is an absolute requirement for their basolateral surface distribution and intracellular localization. PMID- 10452992 TI - Assessment of the role of determinant selection in genetic control of the immune response to insulin in H-2b mice. AB - The immune response to insulin is regulated by MHC class II genes. Immune response (Ir) gene-linked low responsiveness to protein Ags can be mediated by the low affinity of potential antigenic determinants for MHC molecules (determinant selection) or by the influence of MHC on the functional T cell repertoire. Strong evidence exists that determinant selection plays a key role in epitope immunodominance and Ir gene-linked unresponsiveness. However, the actual measurement of relative MHC-binding affinities of all potential peptides derived from well-characterized model Ags under Ir gene regulation has been very limited. We chose to take advantage of the simplicity of the structure of insulin to study the mechanism of Ir gene control in H-2b mice, which respond to beef insulin (BINS) but not pork insulin (PINS). Peptides from these proteins, including the immunodominant A(1-14) determinant, were observed to have similar affinities for purified IAb in binding experiments. Functional and biochemical experiments suggested that PINS and BINS are processed with similar efficiency. The T cell response to synthetic pork A(1-14) was considerably weaker than the response to the BINS peptide. We conclude that the poor immunogenicity of PINS in H-2b mice is a consequence of the T cell repertoire rather than differences in processing and presentation. PMID- 10452993 TI - Phenotypical and functional characterization of the CD8+ T cell repertoire of HLA A2.1 transgenic, H-2KbnullDbnull double knockout mice. AB - Homozygous HLA-A2.1 transgenic H-2KbnullDbnull double knockout (KO) mice were created. Their potential to develop HLA-A2. 1-restricted cytolytic responses was compared with that of their classical transgenic counterparts, which still express H-2Kb, Db molecules. On cell surfaces, both strains express similar amounts of chimeric (alpha 1 alpha 2 domains of human, alpha 3 cytoplasmic domains of mouse) HLA-A2.1 molecules in noncovalent association with mouse beta 2 microglobulin. Compared with mice that are totally deprived of histocompatibility class Ia molecules (H-2KbnullDbnull double KO), the expression of HLA-A2.1 in transgenic/double KO mice resulted in sizeable increase in the periphery of CD8+ T cells with a normally diversified TCR repertoire. A biased education in favor of HLA-A2.1, ascribable to the absence of H-2 class Ia molecules, was evidenced in these transgenic/double KO mice by their improved capacity to mount HLA restricted cytolytic responses, regardless of whether they were virally infected or injected with synthetic epitopic peptide. HLA class I transgenic, H-2 class Ia KO mice should represent useful animal models for the preclinical evaluation of vaccine formulations aiming at the induction of HLA class I-restricted CTL responses. PMID- 10452994 TI - Adjuvant effect of IL-12: conversion of peptide antigen administration from tolerizing to immunizing for CD8+ T cells in vivo. AB - CD8+ T cells from TCR transgenic 2C mice, specific for SIYRYYGL peptide bound to H-2Kb, were adoptively transferred into C57BL/6 recipients to allow monitoring of their location, numbers, and phenotype upon peptide challenge. Recipients were primed by s.c. injection of SIYRYYGL alone or with CFA or IL-12, and the transferred cells then tracked by flow cytometry using the 1B2 mAb specific for the 2C TCR. Peptide alone induced a transient and weak expansion of 1B2+ cells in the draining lymph nodes (DLN) by day 3, but these cells were tolerant to secondary peptide challenge. In contrast, priming with CFA/peptide resulted in a large clonal expansion of 1B2+ cells in DLN by day 3, and the cells exhibited a CD25highCD44high phenotype, blast transformation, and lytic effector function. By day 5, 1B2+ cell numbers decreased in the DLN and increased in the spleen and blood. 1B2+ cells with a memory phenotype persisted through day 60 in the DLN, spleen, and blood and responded to secondary peptide challenge. Immunization with peptide, along with IL-12, mimicked the adjuvant effects of CFA with respect to phenotype, clonal expansion, effector function, and establishment of memory. IL 12 was not unique in providing this adjuvant effect however, since CFA/peptide immunization of IL-12-deficient recipient mice also resulted in 1B2+ T cell activation and clonal expansion. Thus, CFA or IL-12 can enhance Ag-specific CD8+ T cell responses to peptide, demonstrating that an inflammatory cytokine(s) can support activation and prevent tolerance induction. PMID- 10452995 TI - Bromelain, from pineapple stems, proteolytically blocks activation of extracellular regulated kinase-2 in T cells. AB - Recently, it has emerged that extracellular proteases have specific regulatory roles in modulating immune responses. Proteases may act as signaling molecules to activate the Raf-1/extracellular regulated kinase (ERK)-2 pathway to participate in mitogenesis, apoptosis, and cytokine production. Most reports on the role of protease-mediated cell signaling, however, focus on their stimulatory effects. In this study, we show for the first time that extracellular proteases may also block signal transduction. We show that bromelain, a mixture of cysteine proteases from pineapple stems, blocks activation of ERK-2 in Th0 cells stimulated via the TCR with anti-CD3epsilon mAb, or stimulated with combined PMA and calcium ionophore. The inhibitory activity of bromelain was dependent on its proteolytic activity, as ERK-2 inhibition was abrogated by E-64, a selective cysteine protease inhibitor. However, inhibitory effects were not caused by nonspecific proteolysis, as the protease trypsin had no effect on ERK activation. Bromelain also inhibited PMA-induced IL-2, IFN-gamma, and IL-4 mRNA accumulation, but had no effect on TCR-induced cytokine mRNA production. This data suggests a critical requirement for ERK-2 in PMA-induced cytokine production, but not TCR induced cytokine production. Bromelain did not act on ERK-2 directly, as it also inhibited p21ras activation, an effector molecule upstream from ERK-2 in the Raf 1/MEK/ERK-2 kinase signaling cascade. The results indicate that bromelain is a novel inhibitor of T cell signal transduction and suggests a novel role for extracellular proteases as inhibitors of intracellular signal transduction pathways. PMID- 10452996 TI - Signals transduced by CD3epsilon, but not by surface pre-TCR complexes, are able to induce maturation of an early thymic lymphoma in vitro. AB - Development of immature CD4-CD8- (double-negative) thymocytes to the CD4+CD8+ (double-positive) stage is linked to productive rearrangement of the TCRbeta locus by signals transduced through the pre-TCR. However, the mechanism whereby pre-TCR signaling is initiated remains unclear, in part due to the lack of an in vitro model system amenable to both biochemical and genetic analysis. In this study, we establish the thymic lymphoma Scid.adh as such a model system. Scid.adh responds to Ab engagement of surface IL-2Ra (TAC):CD3epsilon molecules (a signaling chimera that mimics pre-TCR signaling in vivo) by undergoing changes in gene expression observed following pre-TCR activation in normal thymocytes. These changes include down-regulation of CD25, recombinase-activating gene (RAG)-1, RAG 2, and pTalpha; and the up-regulation of TCRalpha germline transcripts. We term this complete set of changes in gene expression, in vitro maturation. Interestingly, Scid.adh undergoes only a subset of these changes in gene expression following Ab engagement of the pre-TCR. Our findings make two important points. First, because TAC:CD3epsilon stimulation of Scid.adh induces physiologically relevant changes in gene expression, Scid.adh is an excellent cellular system for investigating the molecular requirements for pre-TCR signaling. Second, Ab engagement of CD3epsilon signaling domains in isolation (TAC:CD3epsilon) promotes in vitro maturation of Scid.adh, whereas engagement of CD3epsilon molecules contained within the complete pre-TCR fails to do so. Our current working hypothesis is that CD3epsilon fails to promote in vitro maturation when in the context of an Ab-engaged pre-TCR because another pre-TCR subunit(s), possibly TCRzeta, qualitatively alters the CD3epsilon signal. PMID- 10452997 TI - Requirement for Shc in TCR-mediated activation of a T cell hybridoma. AB - Engagement of the TCR determines the fate of T cells to activate their functional programs, proliferate, or undergo apoptosis. The intracellular signal transduction pathways that dictate the specific outcome of receptor engagement have only been partially elucidated. The adapter protein, Shc, is involved in cytokine production, mitogenesis, transformation, and apoptosis in different cell systems. We found that Shc becomes phosphorylated on tyrosine residues upon stimulation of the TCR in DO11.10 hybridoma T cells; therefore, we investigated the role of Shc in activation-induced cell death in these cells by creating a series of stably transfected cell lines. Expression of Shc-SH2 (the SH2 domain of Shc) or Shc-Y239/240F (full-length Shc in which tyrosines 239 and 240 have been mutated to phenylalanine) resulted in the inhibition of activation-induced cell death and Fas ligand up-regulation after TCR cross-linking. Expression of wild type Shc or Shc-Y317F had no significant effect. In addition, we found that Shc SH2 and Shc-Y239/240F, but not Shc-Y317F, inhibited phosphorylation of extracellular signal-regulated protein kinase and production of IL-2 after TCR cross-linking. These results indicate an important role for Shc in the early signaling events that lead to activation-induced cell death and IL-2 production after TCR activation. PMID- 10452998 TI - Intranasal exposure to protein antigen induces immunological tolerance mediated by functionally disabled CD4+ T cells. AB - In this study we examined the immunological parameters underlying the natural immunity to inhaled nonpathogenic proteins. We addressed this question by examining the effect of intranasal exposure to OVA in both wild-type mice and mice reconstituted with OVA-TCR transgenic CD4+ T cells. Intranasal administration of OVA induced an initial phase of activation during which CD4+ T cells were capable of proliferating and producing cytokines. Although many of the OVA-specific CD4+ T cells were subsequently depleted from the lymphoid organs, a stable population of such T cells survived but remained refractory to antigenic rechallenge. The unresponsive state was not associated with immune deviation due to selective secretion of Th1- or Th2-type cytokines, and the presence of regulatory CD8+ T cells was not required. Moreover, neutralization of the immunosuppressive cytokines IL-10 and TGF-beta did not abrogate the induction of tolerance. Inhibition of the interaction of T cells with CD86, but not CD80, at the time of exposure to intranasal Ag prevented the development of unresponsiveness, while selective blockade of CTLA-4 had no effect. Our results suggest that intranasal exposure to Ags results in immunological tolerance mediated by functionally impaired CD4+ T cells via a costimulatory pathway that requires CD86. PMID- 10452999 TI - A partially agonistic peptide acts as a selective inducer of apoptosis in CD8+ CTLs. AB - We have analyzed the effect of partially agonistic peptides on the activation and survival of CTL clones specific for a highly immunogenic HLA A11-restricted peptide epitope derived from the EBV nuclear Ag-4. Several analogues with substitutions of TCR contact residues were able to trigger cytotoxic activity without induction of IL-2 mRNA and protein or T cell proliferation. Triggering with these partial agonists in the absence of exogenous IL-2 resulted in down regulation of the cytotoxic potential of the specific CTLs. One analogue selectively triggered apoptosis as efficiently as the original epitope, subdividing the partial agonists into apoptosis-inducing and noninducing ligands. Analysis of early T cell activation events, induction of Ca2+ influx, and acid release did not reveal significant differences between the two types of analogue peptides. These results demonstrate that some partial agonists can dissociate the induction of CTL death from CTL activation. Peptides with such properties may serve as useful tools to study signal transduction pathways in CD8+ lymphocytes and as therapeutic agents modulating natural immune responses. PMID- 10453000 TI - Pre-T cell receptor signals are responsible for the down-regulation of Syk protein tyrosine kinase expression. AB - Thymocyte development proceeds through two critical checkpoints that involve signaling events through two different receptors, the TCR and the pre-TCR. These receptors employ two families of protein tyrosine kinases to propagate their signals, the Src and Syk families. Genetic and biochemical evidence has shown that the Src family kinases are critical for normal T cell maturation. ZAP-70, a Syk family kinase, has similarly been implicated as a critical component in thymocyte development. Although genetic evidence has suggested that Syk is involved during thymocyte development, a definitive study of Syk expression has not been performed. In this paper we report our reanalysis of Syk expression in subpopulations of murine and human thymocytes by intracellular staining and flow cytometry using anti-Syk mAbs. Syk is expressed at increased levels during the stages in which pre-TCR signaling occurs. Furthermore, Syk is down-regulated after the pre-TCR checkpoint has been passed. Syk may play an important role in thymic development during pre-TCR signal transduction. Finally, incomplete down regulation of Syk expression was noted in human thymocytes, offering a possible explanation for the distinct phenotypes of mice and humans deficient in ZAP-70. PMID- 10453001 TI - mdr1a-encoded P-glycoprotein is not required for peripheral T cell proliferation, cytokine release, or cytotoxic effector function in mice. AB - The plasma membrane transport protein P-glycoprotein (P-gp) is expressed by subsets of both CD4+ and CD8+ T cells in mice. The proportion of T cells that express P-gp goes up with age, and the P-gp-expressing subset of the CD4 memory population is hyporesponsive in many in vitro assays. The significance of P-gp expression for T cell function has not been well established, although several reports have suggested that it may promote cytokine export and/or cytotoxic T cell function. To elucidate which T cell functions may require P-gp, we have compared a variety of responses using T cells from wt and P-gp knockout mice. Protein expression and rhodamine-123 efflux studies revealed that peripheral T cells exclusively utilize the mdr1a-encoded isoform rather than the homologous mdr1b or mdr2 isoforms. Comparisons of T cells from mdr1a+/+ and mdr1a-/- mice showed no differences in proliferation or in secretion of IL-2, IL-4, IL-5, IL 10, or IFN-gamma in response to polyclonal stimulation. Moreover, mdr1a-/- T cells produced strong allospecific cytotoxic responses comparable to those of wt T cells. Our results show that P-gp is not a necessary component of peripheral T cell functional responses. Further investigation will be needed to determine the significance of P-gp expression in T lymphocytes. PMID- 10453002 TI - Levels of Ly-49 receptor expression are determined by the frequency of interactions with MHC ligands: evidence against receptor calibration to a "useful" level. AB - Ly-49 receptor expression was studied in NK cells that developed in fully MHC mismatched mixed bone marrow chimeras, in which host and donor MHC ligands were expressed solely on various proportions of hemopoietic cells or on both hemopoietic and nonhemopoietic cells. When hemopoietic cells were the only source of MHC ligand, a strong correlation between the level of down-regulation of Ly 49A, Ly-49C, and Ly-49G2 and the number of hemopoietic cells expressing their MHC ligands was observed on both donor and host NK cells. In some animals with low levels of donor hemopoietic chimerism, NK cells of donor origin expressed Ly-49 receptors at higher levels than was observed in normal mice of the same strain. This unexpected observation is inconsistent with the receptor calibration theory, which states that expression of Ly-49 inhibitory receptors is calibrated to an optimal level to maintain an NK cell repertoire that is sensitive to perturbations in normal class I ligand expression. Our data suggest a model in which Ly-49 receptors down-modulate in accordance with the frequency of their interactions with ligand-bearing cells, rather than a model in which these receptors calibrate to a specific "useful" level in response to ligands present in their environment. PMID- 10453003 TI - The role of CTLA-4 in regulating Th2 differentiation. AB - To examine the role of CTLA-4 in Th cell differentiation, we used two newly generated CTLA-4-deficient (CTLA-4-/-) mouse strains: DO11. 10 CTLA-4-/- mice carrying a class II restricted transgenic TCR specific for OVA, and mice lacking CTLA-4, B7.1 and B7.2 (CTLA-4-/- B7.1/B7.2-/- ). When purified naive CD4+ DO11.10 T cells from CTLA-4-/- and wild-type mice were primed and restimulated in vitro with peptide Ag, CTLA-4-/- DO11.10 T cells developed into Th2 cells, whereas wild type DO11.10 T cells developed into Th1 cells. Similarly, when CTLA-4-/- CD4+ T cells from mice lacking CTLA-4, B7. 1, and B7.2 were stimulated in vitro with anti-CD3 Ab and wild-type APC, these CTLA-4-/- CD4+ T cells produced IL-4 even during the primary stimulation, whereas CD4+ cells from B7.1/B7.2-/- mice did not produce IL-4. Upon secondary stimulation, CD4+ T cells from CTLA-4-/- B7.1/B7.2-/ mice secreted high levels of IL-4, whereas CD4+ T cells from B7.1/B7.2-/- mice produced IFN-gamma. In contrast to the effects on CD4+ Th differentiation, the absence of CTLA-4 resulted in only a modest effect on T cell proliferation, and increased proliferation of CTLA-4-/- CD4+ T cells was seen only during secondary stimulation in vitro. Administration of a stimulatory anti-CD28 Ab in vivo induced IL-4 production in CTLA-4-/- B7.1/B7.2-/- but not wild-type mice. These studies demonstrate that CTLA-4 is a critical and potent inhibitor of Th2 differentiation. Thus, the B7-CD28/CTLA-4 pathway plays a critical role in regulating Th2 differentiation in two ways: CD28 promotes Th2 differentiation while CTLA-4 limits Th2 differentiation. PMID- 10453004 TI - Lipopolysaccharide induces in macrophages the synthesis of the suppressor of cytokine signaling 3 and suppresses signal transduction in response to the activating factor IFN-gamma. AB - The goal of this study was to investigate how bacterial LPS affects macrophage responsiveness to the activating factor IFN-gamma. Pretreatment of macrophages with LPS for <2 h increased the transcriptional response to IFN-gamma. In contrast, simultaneous stimulation with IFN-gamma and LPS, or pretreatment with LPS for >4 h, suppressed Stat1 tyrosine 701 phosphorylation, dimerization, and transcriptional activity in response to IFN-gamma. Consistently, the induction of MHCII protein by IFN-gamma was antagonized by LPS pretreatment. Neutralizing Abs to IL-10 were without effect on LPS-mediated suppression of Stat1 activation. Decreased IFN-gamma signal transduction after LPS treatment corresponded to a direct induction of suppressor of cytokine signaling3 (SOCS3) mRNA and protein. Under the same conditions socs1, socs2, and cis genes were not transcribed. In transfection assays, SOCS3 was found to suppress the transcriptional response of macrophages to IFN-gamma. A causal link of decreased IFN-gamma signaling to SOCS3 induction was also suggested by the LPS-dependent reduction of IFN-gamma-mediated Janus kinase 1 (JAK1) activation. Further consistent with inhibitory activity of SOCS3, LPS also inhibited the JAK2-dependent activation of Stat5 by GM-CSF. Our results thus link the deactivating effect of chronic LPS exposure on macrophages with its ability to induce SOCS3. PMID- 10453005 TI - Differentiation of NK1.1+, Ly49+ NK cells from flt3+ multipotent marrow progenitor cells. AB - To delineate factors involved in NK cell development, we established an in vitro system in which lineage marker (Lin)-, c-kit+, Sca2+ bone marrow cells differentiate into lytic NK1.1+ but Ly49- cells upon culture in IL-7, stem cell factor (SCF), and flt3 ligand (flt3L), followed by IL-15 alone. A comparison of the ability of IL-7, SCF, and flt3L to generate IL-15-responsive precursors suggested that NK progenitors express the receptor for flt3L. In support of this, when Lin-, c-kit+, flt3+ or Lin-, c-kit+, flt3- progenitors were utilized, 3-fold more NK cells arose from the flt3+ than from the flt3- progenitors. Furthermore, NK cells that arose from flt3- progenitors showed an immature NK1.1dim, CD2-, c kit+ phenotype as compared with the more mature NK1.1bright, CD2+/-, c-kit- phenotype displayed by NK cells derived from flt3+ progenitors. Both progenitors, however, gave rise to NK cells that were Ly49 negative. To test the hypothesis that additional marrow-derived signals are necessary for Ly49 expression on developing NK cells, flt3+ progenitors were grown in IL-7, SCF, and flt3L followed by culture with IL-15 and a marrow-derived stromal cell line. Expression of Ly49 molecules, including those of which the MHC class I ligands were expressed on the stromal or progenitor cells, as well as others of which the known ligands were absent, was induced within 6-13 days. Thus, we have established an in vitro system in which Ly49 expression on developing NK cells can be analyzed and possibly experimentally manipulated. PMID- 10453006 TI - Analysis of a 26-kb region linked to the Mhc in zebrafish: genomic organization of the proteasome component beta/transporter associated with antigen processing-2 gene cluster and identification of five new proteasome beta subunit genes. AB - Sequencing of zebrafish (Danio rerio) bacterial artificial chromosome and P1 artificial chromosome genomic clone fragments and of cDNA clones has led to the identification of five new loci coding for beta subunits of proteasomes (PSMB). Together with the four genes identified previously, nine PSMB genes have now been defined in the zebrafish. Six of the nine genes reside in the zebrafish MHC (Mhc) class I region, four of them reside in a single cluster closely associated with TAP2 on a 26-kb long genomic fragment, and two reside at some distance from the fragment. In addition to homologues of the human genes PSMB5 through PSMB9, two new genes, PSMB11 and PSMB12, have been found for which there are no known corresponding genes in humans. The new genes reside in the PSMB cluster in the Mhc. Homology and promoter region analysis suggest that the Mhc-associated genes might be inducible by IFN-gamma. The zebrafish class I region contains representatives of three phylogenetically distinguishable groups of PSMB genes, X, Y, and Z. It is proposed that these genes were present in the ancestral PSMB region before Mhc class I genes became associated with it. PMID- 10453007 TI - Human endothelial cells augment early CD40 ligand expression in activated CD4+ T cells through LFA-3-mediated stabilization of mRNA. AB - Human endothelial cells (EC) augment CD40 ligand (CD40L) expression on PHA activated CD4+ T cells at early times (e.g., 4-6 h). Fixed EC, devoid of mRNA, are comparable to living EC in their capacity to augment early CD40L expression on CD4+ T cells. Fixed EC increase T cell mRNA expression of both IL-2 and CD40L compared with PHA alone at 6 h. EC are unable to increase the rate of transcription of CD40L compared with PHA alone as measured with a promoter reporter gene, although they do increase transcription of an IL-2 promoter reporter gene. Fixed EC prolong the half-life of CD40L mRNA >2-fold. Inclusion of anti-human LFA-3 (CD58) mAb or pretreatment of EC with an LFA-3 antisense oligonucleotide blocks EC-induced increases in CD40L expression, whereas mAb to ICAM-1 or pretreatment with ICAM-1 antisense oligonucleotide does not. Moreover, mAb to LFA-3 reverses the capacity of EC to prolong the half-life of CD40L mRNA, whereas mAb to ICAM-1, even in combination with mAb to ICAM-2, does not. We conclude that EC use LFA-3 to increase early CD40L protein expression on newly activated CD4+ T cells by stabilizing CD40L mRNA. PMID- 10453009 TI - T cell development in PU.1-deficient mice. AB - These studies address the role of PU.1 in T cell development through the analysis of PU.1-/- mice. We show that the majority of PU.1-/- thymocytes are blocked in differentiation prior to T cell commitment, and contain a population of thymocyte progenitors with the cell surface phenotype of CD44+, HSAbright, c-kitint, Thy-1 , CD25-, Sca-1-, CD4-, and CD8-. These cells correspond in both number and cell surface phenotype with uncommitted thymocyte progenitors found in wild-type fetal thymus. RT-PCR analysis demonstrated that PU.1 is normally expressed in this early progenitor population, but is down-regulated during T cell commitment. Rare PU.1-/- thymi, however, contained small numbers of thymocytes expressing markers of T cell commitment. Furthermore, almost 40% of PU.1-/- thymi placed in fetal thymic organ culture are capable of T cell development. Mature PU. 1-/- thymocytes generated during organ culture proliferated and produced IL-2 in response to stimulation through the TCR. These data demonstrate that PU.1 is not absolutely required for T cell development, but does play a role in efficient commitment and/or early differentiation of most T progenitors. PMID- 10453008 TI - A composite C/EBP binding site is essential for the activity of the promoter of the IL-3/IL-5/granulocyte-macrophage colony-stimulating factor receptor beta c gene. AB - The common beta-chain (beta c) is the main signaling component of the heterodimeric receptors for IL-3, IL-5, and GM-CSF and is primarily expressed on myeloid cells. The proximal beta c promoter is regulated by GGAA binding proteins, including PU.1, a hemopoietic specific member of the Ets family. However, it is not likely that PU.1 alone accounts for the myeloid-restricted expression of the beta c subunit. Here we describe the identification of a C/EBP binding enhancer that is located 2 kb upstream of the transcription start site. The enhancer contains two elements that bind C/EBP alpha and -beta in U937 cells, while C/EBP epsilon is also bound in extracts of HL-60 cells. Importantly, deletion of the enhancer or mutation of either of one of the C/EBP sites results in a complete loss of promoter activity in cell lines as well as in primary cells, showing the importance of C/EBP members in beta c gene activation. We further show that PU.1 has to cooperate with C/EBP proteins to induce beta c transcription. Since the beta c is already expressed on CD34+ cells, these results demonstrate that both C/EBP and PU.1 are not only important for the myeloid-specific gene regulation at later stages of myeloid differentiation. PMID- 10453010 TI - Attenuation of MHC class II expression in macrophages infected with Mycobacterium bovis bacillus Calmette-Guerin involves class II transactivator and depends on the Nramp1 gene. AB - The natural resistance associated macrophage protein 1 (Nramp1) gene determines the ability of murine macrophages to control infection with a group of intracellular pathogens, including Salmonella typhimurium, Leishmania donovani, and Mycobacterium bovis bacillus Calmette-Guerin (BCG). The expression of the resistant allele of the Nramp1 gene in murine macrophages is associated with a more efficient expression of several macrophage activation-associated genes, including class II MHC loci. In this study, we investigated the molecular mechanisms involved in IFN-gamma-induced MHC class II expression in three types of macrophages: those expressing a wild-type allele of the Nramp1 gene (B10R and 129/Mphi), those carrying a susceptible form of the Nramp1 gene (B10S), and those derived from 129-Nramp1-knockout mice (129/Nramp1-KO). Previously, we published results showing that Ia protein expression is significantly higher in the IFN gamma-induced B10R macrophages, compared with its susceptible counterpart. In this paper, we also show that the higher expression of Ia protein in B10R cells is associated with higher I-Abeta mRNA expression, which correlates with a higher level of IFN-gamma-induced phosphorylation of the STAT1-alpha protein and subsequently with elevated expression of class II transactivator (CIITA) mRNA, compared with B10S. Furthermore, we demonstrate that the infection of macrophages with M. bovis BCG results in a down-regulation of CIITA mRNA expression and, consequently, in the inhibition of Ia induction. Therefore, our data explain, at least in part, the molecular mechanism involved in the inhibition of I-Abeta gene expression in M. bovis BCG-infected macrophages activated with IFN-gamma. PMID- 10453011 TI - Lymphoid transcription of the murine CD21 gene is positively regulated by histone acetylation. AB - Intronic transcriptional control sequences influence the cell- and tissue specific expression of the CD21 gene. The interactions of such intronic control sequences, which are physically separated from the gene's promoters, suggest that factors that alter chromatin structure might be influential in this process. Accordingly, we analyzed the effect of histone acetylation on the expression of CD21 in nonexpressing T and B lymphocytes, respectively. The acetylase inhibitors sodium butyrate and trichostatin A were used to create hyperacetylated histones. The CD21 gene was specifically activated in the previously transcriptionally quiescent cells in a time- and dose-dependent fashion: the expression of a number of other genes was not influenced. These data suggest a model of cell-type specific deacetylase activity that serves to repress gene transcription when present and active. PMID- 10453012 TI - A role for the transmembrane domain in the trimerization of the MHC class II associated invariant chain. AB - MHC class II and invariant chain (Ii) associate early in biosynthesis to form a nonameric complex. Ii first assembles into a trimer and then associates with three class II alphabeta heterodimers. Although the membrane-proximal region of the Ii luminal domain is structurally disordered, the C-terminal segment of the luminal domain is largely alpha-helical and contains a major interaction site for the Ii trimer. In this study, we show that the Ii transmembrane domain plays an important role in the formation of Ii trimers. The Ii transmembrane domain contains an unusual patch of hydrophilic residues near the luminal interface. Substitution of these polar residues with nonpolar amino acids resulted in a decrease in the efficiency of Ii trimerization and subsequent class II association. Moreover, N-terminal fragments of Ii were found to trimerize independently of the luminal alpha-helical domain. Progressive C-terminal truncations mapped a homotypic association site to the first 80 aa of Ii. Together, these results implicate the Ii transmembrane domain as a site of trimer interaction that can play an important role in the initiation of trimer formation. PMID- 10453013 TI - Differential expression and distinct functions of IFN regulatory factor 4 and IFN consensus sequence binding protein in macrophages. AB - IFN regulatory factor 4 (IRF4) and IFN consensus sequence binding protein (ICSBP) are highly homologous members of the growing family of IRF proteins. ICSBP expression is restricted to lymphoid and myeloid cells, whereas IRF4 expression has been reported to be lymphoid-restricted. We present evidence that primary murine and human macrophages express IRF4, thereby extending its range of expression to myeloid cells. Here, we provide a comparative analysis of IRF4 and ICSBP expression and function in distinct cell types. These IRF proteins can form specific complexes with the Ets-like protein PU.1, and can activate transcription via binding to PU.1/IRF composite sequences. EMSA analysis revealed that murine macrophages contained both IRF4/PU.1 and ICSBP/PU.1 complexes, analogous to B cells. Over-expression of ICSBP in these macrophages activated transcription of a PU.1/IRF-dependent promoter, whereas over-expression of IRF4 had no effect on this promoter. In contrast, over-expression of either IRF4 or ICSBP in both macrophages and NIH-3T3 fibroblasts suppressed transcription of the PU.1 independent H-2Ld MHC class I promoter. In NIH-3T3 fibroblasts, IRF4 and ICSBP also synergized with exogenous PU.1 to activate transcription of a PU.1/IRF dependent promoter. Furthermore, both IRF4 and ICSBP activated transcription of the IL-1beta promoter in both cell types. While this promoter is PU.1-dependent, it lacks any known PU.1/IRF composite binding sites. Synergistic activation of the IL-1beta promoter by these IRF proteins and PU.1 was found to require PU.1 serine 148. Together, these data demonstrate that IRF4 and ICSBP are dichotomous regulators of transcription in macrophages. PMID- 10453014 TI - TCR reactivity in human nickel allergy indicates contacts with complementarity determining region 3 but excludes superantigen-like recognition. AB - Nickel is the most common inducer of contact sensitivity in humans. We previously found that overrepresentation of the TCRBV17 element in Ni-induced CD4+ T cell lines of Ni-allergic patients relates to the severity of the disease. Amino acid sequences of these beta-chains suggested hypothetical contact points for Ni2+ ions in complementarity-determining region (CDR) 1 and CDR3. To specifically address the molecular requirements for Ni recognition by TCR, human TCR alpha- and beta-chains of VB17+ Ni-reactive T cell clones were functionally expressed together with the human CD4 coreceptor in a mouse T cell hybridoma. Loss of CD4 revealed complete CD4 independence for one of the TCR studied. Putative TCR/Ni contact points were tested by pairing of TCR chains from different clones, also with different specificity. TCRBV17 chains with different J regions, but similar CDR3 regions, could be functionally exchanged. Larger differences in the CDR3 region were not tolerated. Specific combinations of alpha- and beta-chains were required, excluding a superantigen-like activation by Ni. Mutation of amino acids in CDR1 of TCRBV17 did not affect Ag recognition, superantigen activation, or HLA restriction. In contrast, mutation of Arg95 or Asp96, conserved in many CDR3B sequences of Ni-specific, VB17+ TCR, abrogated Ni recognition. These results define specific amino acids in the CDR3B region of a VB17+ TCR to be crucial for human nickel recognition. CD4 independence implies a high affinity of such receptor types for the Ni/MHC complex. This may point to a dominant role of T cells bearing such receptors in the pathology of contact dermatitis. PMID- 10453015 TI - Biased VH gene usage in early lineage human B cells: evidence for preferential Ig gene rearrangement in the absence of selection. AB - Certain VH genes are predominantly expressed in mature B cells. We hypothesized that several, mutually nonexclusive VH-dependent mechanisms operating at distinct stages during B cell development may be responsible for overrepresentation of these VH genes. In the present study, we have assessed whether one of the mechanisms involves preferential rearrangement at the pro-B cell stage. The frequency of individual VH4 and VH3 genes in rearrangement libraries from FACS purified human CD34+/CD19+ pro-B and CD34-/CD19+ pre-B cells was assessed. The in frame and out-of-frame rearrangements from both cell populations were analyzed using a high resolution PAGE system. The frequencies of individual VH gene segments among out-of-frame rearrangements from pro-B cells were determined, because these frequencies should reflect only processes before the translation of the mu-heavy chain and should not be biased by selection mechanisms. Our results demonstrate that, at the pro-B cell stage, the V4-34, V4-39, and V4-59 gene segments are the most frequently rearranged VH4 family genes, and the V3-23 and V3-30 gene segments are the most frequently rearranged VH3 family genes. This finding suggests that the predominant expression of these VH genes in peripheral mature B cells is determined to a significant degree by their preferential rearrangement during V-DJ recombination. PMID- 10453016 TI - Immunization with Treponema pallidum outer membrane vesicles induces high-titer complement-dependent treponemicidal activity and aggregation of T. pallidum rare outer membrane proteins (TROMPs). AB - The purpose of this study was to determine whether immunization with purified outer membrane vesicles (OMV) from Treponema pallidum (T.p. ) could elicit Abs capable of killing this organism. It is well established that the immunization of rabbits or mice with killed T.p. or with recombinant T.p. Ags has failed to generate serum killing activity comparable with that of infection-derived immunity. Because of the small amount of T.p. OMV obtainable, a single mouse was immunized with purified OMV. The mouse anti-OMV serum and infection-derived immune rabbit serum (IRS) were compared by reactivities on two-dimensional T.p. immunoblots and by the T.p. immobilization test, a complement-dependent killing assay. Whereas IRS detected >40 Ags, the anti-OMV serum identified only 6 Ags corresponding to proteins identified previously in the outer membrane. T.p. immobilization testing showed that IRS had a 100% killing titer of 1:44 and a 50% killing titer of 1:662. By comparison, the mouse anti-OMV serum had a significantly greater 100% killing titer of 1:1,408 and a 50% killing titer of 1:16,896. Absorption of the anti-OMV serum to remove Ab against outer membrane associated lipoproteins did not change the 100% killing titer. Freeze-fracture analysis of T.p. incubated in IRS or anti-OMV serum showed that T.p. rare membrane-spanning outer membrane proteins were aggregated. This is the first demonstration of high-titer killing Abs resulting from immunization with defined T.p. molecules; our study indicates that the targets for these Abs are T. p. rare outer membrane proteins. PMID- 10453017 TI - Partial IL-10 inhibition of the cell-mediated immune response in chronic beryllium disease. AB - Chronic beryllium disease (CBD) provides a human disorder in which to study the delayed type IV hypersensitivity response to persistent Ag that leads to noncaseating pulmonary granuloma formation. We hypothesized that, in CBD, failure of IL-10 to modulate the beryllium-specific, cell-mediated immune response would result in persistent, maximal cytokine production and T lymphocyte proliferation, thus contributing to the development of granulomatous lung disease. To test this hypothesis, we used bronchoalveolar lavage cells from control and CBD subjects to evaluate the beryllium salt-specific production of endogenous IL-10 and the effects of exogenous human rIL-10 (rhIL-10) on HLA expression, on the production of IL-2, IFN-gamma, and TNF-alpha, and on T lymphocyte proliferation. Our data demonstrate that beryllium-stimulated bronchoalveolar lavage cells produce IL-10, and the neutralization of endogenous IL-10 does not increase significantly cytokine production, HLA expression, or T lymphocyte proliferation. Second, the addition of excess exogenous rhIL-10 partially inhibited the beryllium-stimulated production of IL-2, IFN-gamma, and TNF-alpha; however, we measured no change in T lymphocyte proliferation or in the percentage of alveolar macrophages expressing HLA-DP. Interestingly, beryllium salts interfered with an IL-10-stimulated decrease in the percentage of alveolar macrophages expressing HLA-DR. We conclude that, in the CBD-derived, beryllium-stimulated cell-mediated immune response, low levels of endogenous IL-10 have no appreciable effect; exogenous rhIL-10 has a limited effect on cytokine production and no effect on T lymphocyte proliferation or HLA expression. PMID- 10453018 TI - Partial agonist effect influences the CTL response to a heterologous dengue virus serotype. AB - Activation of dengue serotype-cross-reactive memory CTL during secondary dengue virus (DV) infection is thought to be important in the pathogenesis of dengue hemorrhagic fever. To model this effect, we studied the CTL responses to DV types 2 (D2V) and 3 (D3V) in PBMC from an individual previously infected with D3V. DV specific CD8+ CTL from this donor recognized two HLA-B62-restricted epitopes on the NS3 protein, aa 71-79 (SVKKDLISY) and 235-243 (AMKGLPIRY). Both D3V-specific and D2V/D3V-cross-reactive CTL clones were detected for each epitope; all D2V reactive CTL clones could lyse D2V-infected autologous cells. CTL responses to both epitopes were detected in bulk cultures stimulated with D3V, but PBMC stimulated with D2V recognized only the 235-243 epitope. IFN-gamma enzyme-linked immunospot assay showed that the D2V (71-79) peptide (DVKKDLISY) did not efficiently activate T cells. Analysis of a CTL clone suggests that the D2V (71 79) peptide acts as a partial agonist, able to sensitize target cells for lysis and inducing only minimal proliferation at high concentrations. These results suggest that variant peptide sequences present in the heterologous DV serotype can influence the CTL response in vivo during secondary DV infection. PMID- 10453019 TI - Role of complement component C1q in the IgG-independent opsonophagocytosis of group B streptococcus. AB - We investigated the role of complement component C1q in the IgG-independent opsonophagocytosis of type III group B Streptococcus (GBS) by peripheral blood leukocytes. We report that C1q binds to type III GBS both in normal human serum deficient in IgG specific for type III capsular polysaccharide and in a low-ionic strength buffer. The dissociation constant Kd ranged from 2.0 to 5.5 nM, and the number of binding sites Bmax ranged from 630 to 1360 molecules of C1q per bacterium (CFU). An acapsular mutant strain of GBS bound C1q even better than the wild type, indicating that the polysaccharide capsule is not the receptor for C1q. In serum, binding of C1q to GBS was associated with activation of the classical complement pathway. However, normal human serum retained significant opsonic activity after complete depletion of C1q, suggesting that the serum contains a molecule that is able to replace C1q in opsonization and/or complement activation. Mannan-binding lectin, known to share some functions with C1q, appeared not to be involved, since its depletion from serum had little effect on opsonic activity. Excess soluble C1q or its collagen-like fragment inhibited phagocytosis mediated by normal human serum, suggesting that C1q may compete with other opsonins for binding to receptor(s) on phagocytes. We conclude that, although C1q binds directly to GBS, C1q binding is neither necessary nor sufficient for IgG-independent opsonophagocytosis. The results raise the possibility that additional unknown serum factor(s) may contribute to opsonization of GBS directly or via a novel mechanism of complement activation. PMID- 10453020 TI - Enhanced antigen transport across rat tracheal epithelium induced by sensitization and mast cell activation. AB - Ag challenge to the apical surface of tracheal epithelium results in a rapid ion secretory response due to the activation of mast cells. The aim of this study was to examine the impact of sensitization and specific Ag challenge on the timing, route, and quantity of Ag transported across tracheal epithelium. After sensitization of rats to a model protein, HRP, tracheal tissues were excised and mounted in Ussing chambers. Tracheas from HRP-sensitized rats, but not naive or OVA-sensitized rats, responded to apical HRP challenge with a rise in short circuit current (beginning at approximately 2 min). Photomicrographs of tissues fixed at 2 min showed that initial transepithelial HRP transport occurred via endosomes and was significantly enhanced in HRP-sensitized rats compared with both control groups. In addition, nonciliated cells, the proportion of which increased after sensitization, contained significantly more HRP than ciliated cells. The hypersensitivity response occurred only in HRP-sensitized and challenged rats and was associated with increased conductance of tracheal epithelium and overall flux of HRP across the tissue. This increased flux of Ag and elevated conductance was not observed in mast cell-deficient Ws/Ws rats. Photomicrographs of tissues fixed 90 min after challenge also showed HRP in the paracellular spaces between adjacent epithelial cells. We conclude that sensitization increases uptake of specific Ag initially via an endosomal transcellular pathway across tracheal epithelium and that, after the hypersensitivity reaction, mast cell-dependent recruitment of the paracellular pathway further augments Ag influx into airway tissue. PMID- 10453021 TI - Role of IL-10 in a neonatal mouse listeriosis model. AB - This study was undertaken to test the hypothesis that altered IL-10 production plays a role in the increased susceptibility of neonates to listeriosis. Plasma IL-10 levels were measured in neonatal and adult mice at various times after infection with Listeria monocytogenes. Relative to adults, neonatal mice had markedly increased IL-10 levels early in the course of infection with Listeria using a 90% lethal dose. Higher neonatal IL-10 responses were also observed after injecting adults and pups with equal doses of killed organisms. Splenic macrophages from neonates produced higher IL-10 levels than those of adults after in vitro stimulation with killed bacteria, confirming in vivo observations. Moreover, IL-10 blockade had differential effects in neonates and adults infected with live Listeria. In adult mice, anti-IL-10 Abs decreased bacterial burden early in the course of infection, but were no longer effective at 6 days or later after challenge. In the pups, however, the same treatment had beneficial effects both early and late during infection and resulted in increased survival. Collectively, our data suggest that an overproduction of IL-10 by macrophages may at least partially explain the increased susceptibility of neonates to listeriosis, and provide further evidence that cytokine production is different in adults and neonates. PMID- 10453022 TI - Identification of natural antigenic peptides of a human gastric signet ring cell carcinoma recognized by HLA-A31-restricted cytotoxic T lymphocytes. AB - Peptides of human melanomas recognized by CD8+ CTLs have been identified, but the nature of those of nonmelanoma tumors remains to be elucidated. Previously, we established a gastric signet ring cell carcinoma HST-2 and HLA-A31 (A*31012) restricted autologous CTL clone, TcHST-2. In the present study, we determined the natural antigenic peptides of HST-2 cells. The purified preparation of acid extracted Ags was submitted to the peptide sequencer, and one peptide, designated F4.2 (Tyr-Ser-Trp-Met-Asp-Ile-Ser-Cys-Trp-Ile), appeared to be immunogenic. To confirm the antigenicity of F4.2 further, we constructed an expression minigene vector (pF4.2ss) coding adenovirus E3, a 19-kDa protein signal sequence plus F4.2. An introduction of pF4.2ss minigene to HST-2 and HLA-A31(+) allogeneic tumor cells clearly enhanced and induced the TcHST-2 reactivity, respectively. Furthermore, when synthetic peptides of F4.2 C-terminal-deleted peptides were pulsed to HST-2 cells, F4.2-9 (nonamers), but not F4.2-8 or F4.2-7 (octamer or heptamer, respectively), enhanced the reactivity of TcHST-2, suggesting that the N-terminal ninth Trp might be a T cell epitope. This was confirmed by lack of antigenicity when using synthetic substituted peptides as well as minigenes coding F4.2 variant peptides with Ala or Arg at the ninth position of F4.2. Meanwhile, it was indicated that the sixth position Ile was critically important for the binding to HLA-A31 molecules. Thus, our data indicate that F4.2 may work as an HLA-A31-restricted natural antigenic peptide recognized by CTLs. PMID- 10453023 TI - Lipopolysaccharide binding protein and serum amyloid A secretion by human intestinal epithelial cells during the acute phase response. AB - The acute phase proteins LPS binding protein (LBP) and serum amyloid A (SAA) are produced by the liver and are present in the circulation. Both proteins have been shown to participate in the immune response to endotoxins. The intestinal mucosa forms a large surface that is continuously exposed to these microbial products. By secretion of antimicrobial and immunomodulating agents, the intestinal epithelium contributes to the defense against bacteria and their products. The aim of this study was to explore the influence of the inflammatory mediators TNF alpha, IL-6, and IL-1beta on the release of LBP and SAA by intestinal epithelial cells (IEC). In addition, the induction of LBP and SAA release by cell lines of intestinal epithelial cells and hepatic cells was compared. The data obtained show that in addition to liver cells, IEC also expressed LBP mRNA and released bioactive LBP and SAA upon stimulation. Regulation of LBP and SAA release by IEC and hepatocytes was typical for class 1 acute phase proteins, although differences in regulation between the cell types were observed. Endotoxin did not induce LBP and SAA release. Glucocorticoids were demonstrated to strongly enhance the cytokine-induced release of LBP and SAA by IEC, corresponding to hepatocytes. The data from this study, which imply that human IEC can produce LBP and SAA, suggest a role for these proteins in the local defense mechanism of the gut to endotoxin. Furthermore, the results demonstrate that tissues other than the liver are involved in the acute phase response. PMID- 10453024 TI - Stem cell factor is localized in, released from, and cleaved by human mast cells. AB - Stem cell factor (SCF) is the most important cytokine regulating human mast cell growth and functions. The immunogold technique showed SCF in the secretory granules of skin mast cells and in lung parenchymal mast cells (HLMC). Immunoreactive SCF (iSCF) was detected in cell lysates of HLMC, but not in basophils; iSCF and histamine were detected in supernatants of HLMC 3 min after challenge with anti-FcepsilonRI or anti-IgE, and iSCF in supernatants rapidly declined after 30 min, whereas histamine remained unchanged for 120 min. HPLC and electrospray mass spectrometry (ES/MS) analysis of recombinant human SCF1-166 (18,656. 9 +/- 0.9 Da) treated with chymase showed a polypeptide of 17,977.1 +/- 0.6 Da and a minor component of 697.4 +/- 0.1 Da generated by specific cleavage at Phe159. SCF1-166 and SCF1-159 similarly activated HLMC, potentiated anti-IgE induced activation of these cells, and stimulated HLMC chemotaxis. SCF159-166 had no effect on mast cells. Western blot analysis of supernatants of anti-IgE activated HLMC incubated with recombinant human SCF1-166 showed that SCF1-166 was rapidly cleaved to SCF1-159 and SCF1-144. Experiments with supernatants of anti IgE-activated HLMC incubated with SCF1-166 yielded similar results. In conclusion, SCF is stored in mast cell secretory granules and is immunologically released by human mast cells. SCF1-166 is rapidly and specifically cleaved to SCF1-159 by chymase, which retains its biological effect on mast cells. SCF is also cleaved by other proteases to several SCF species whose possible biological activities remain to be established. PMID- 10453025 TI - Mice with a targeted mutation in lymphotoxin-alpha exhibit enhanced tumor growth and metastasis: impaired NK cell development and recruitment. AB - Mice deficient in lymphotoxin (LT)-alpha lack peripheral lymph nodes and Peyer's patches and have profound defects in development of follicular dendritic cell networks, germinal center formation, and T/B cell segregation in the spleen. Although LTalpha is known to be expressed by NK cells as well as T and B lymphocytes, the requirement of LTalpha for NK cell functions is largely unknown. To address this issue, we have assessed NK cell functions in LTalpha-deficient mice by evaluating tumor models with known requirements for NK cells to control their growth and metastasis. Syngeneic B16F10 melanoma cells inoculated s.c. grew more rapidly in LTalpha-/- mice than in the wild-type littermates, and the formation of experimental pulmonary metastases was significantly enhanced in LTalpha-/- mice. Although LTalpha-/- mice exhibited almost a normal total number of NK cells in spleen, they showed an impaired recruitment of NK cells to lung and liver. Additionally, lytic NK cells were not efficiently produced from LTalpha-/- bone marrow cells in vitro in the presence of IL-2 and IL-15. These data suggest that LTalpha signaling may be involved in the maturation and recruitment of NK cells and may play an important role in antitumor surveillance. PMID- 10453026 TI - Respiratory syncytial virus stimulates neutrophil degranulation and chemokine release. AB - Neutrophil infiltration of the airways is a common finding in respiratory syncytial virus (RSV) bronchiolitis. Neutrophil-derived chemokines and neutrophil granule contents can cause further inflammation, hyperresponsiveness, and damage of the airways. In this study, peripheral blood neutrophils incubated with RSV (multiplicity of infection (MOI) = 10) induced IL-8, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and myeloperoxidase (MPO) release. In contrast, LPS induced only chemokine but not MPO release. RSV-induced chemokine and MPO release was noncytotoxic as assessed by trypan blue exclusion. The mechanism of RSV-induced chemokine release was shown to be transcription dependent since cytokine mRNA synthesis was increased with RSV stimulation and the process was inhibited by actinomycin-D. In addition, the effect of dexamethasone (dex) on mediator release was also studied. Dex significantly inhibited chemokine release but did not inhibit MPO release. The mechanism of inhibition of the release of these chemokines is probably posttranscriptional since the mRNA synthesis was not inhibited by dex. We conclude that the release of chemokines (IL-8, MIP-1alpha, MIP-1beta) and granule enzymes (MPO) by RSV-stimulated neutrophils may contribute to the pulmonary pathology in RSV bronchiolitis. These in vitro findings showing that dex failed to consistently inhibit all the RSV-induced release of neutrophil inflammatory mediators may explain the variable efficacy of corticosteroids in the treatment of RSV bronchiolitis. PMID- 10453027 TI - Altered immune responses and susceptibility to Leishmania major and Staphylococcus aureus infection in IL-18-deficient mice. AB - IL-18, formerly designated IFN-inducing factor, is a novel cytokine produced by activated macrophages. It synergizes with IL-12 in the induction of the development of Th1 cells and NK cells. To define the biological role of IL-18 in vivo, we have constructed a strain of mice lacking IL-18. Homozygous IL-18 knockout (-/-) mice are viable, fertile, and without evident histopathologic abnormalities. However, in contrast to the heterozygous (+/-) or wild-type (+/+) mice, which are highly resistant to the infection of the protozoan parasite Leishmania major, the IL-18-/- mice are uniformly susceptible. The infected IL-18 /- mice produced significantly lower levels of IFN-gamma and larger amounts of IL 4 compared with similarly infected +/- and +/+ mice. In contrast, when infected with the extracellular Gram-positive bacteria Staphylococcus aureus, the IL-18-/- mice developed markedly less septicemia than similarly infected wild-type (+/+) mice. However, the mutant mice developed significantly more severe septic arthritis than the control wild-type mice. This was accompanied by a reduction in the levels of Ag-induced splenic T cell proliferation, decreased IFN-gamma and TNF-alpha synthesis, but increased IL-4 production by the mutant mice compared with the wild-type mice. These results therefore provide direct evidence that IL 18 is not only essential for the host defense against intracellular infection, but it also plays a critical role in regulating the synthesis of inflammatory cytokines, and therefore could be an important target for therapeutic intervention. PMID- 10453028 TI - Inhibition of murine neutrophil recruitment in vivo by CXC chemokine receptor antagonists. AB - In this study, we have examined the ability of chemokine receptor antagonists to prevent neutrophil extravasation in the mouse. Two murine CXC chemokines, macrophage-inflammatory protein (MIP)-2 and KC, stimulated the accumulation of leukocytes into s.c. air pouches, although MIP-2 was considerably more potent. The leukocyte infiltrate was almost exclusively neutrophilic in nature. A human CXC chemokine antagonist, growth-related oncogene (GRO)-alpha(8-73), inhibited calcium mobilization induced by MIP-2, but not by platelet-activating factor in leukocytes isolated from the bone marrow, indicating that this antagonist inhibits MIP-2 activity toward murine leukocytes. Pretreatment of mice with GROalpha(8-73) inhibited, in a dose-dependent manner, the MIP-2-induced influx of neutrophils to levels that were not significantly different from control values. Moreover, this antagonist was also effective in inhibiting the leukocyte recruitment induced by TNF-alpha, LPS, and IL-1beta. Leukocyte infiltration into the peritoneal cavity in response to MIP-2 was also inhibited by prior treatment of mice with GROalpha(8-73) or the analogue of platelet factor 4, PF4(9-70). The results of this study indicate 1) that the murine receptor for MIP-2 and KC, muCXCR2, plays a major role in neutrophil recruitment to s.c. tissue and the peritoneal cavity in response to proinflammatory agents and 2) that CXCR2 receptor antagonists prevent acute inflammation in vivo. PMID- 10453030 TI - Acceleration and increased severity of collagen-induced arthritis in P-selectin mutant mice. AB - P-selectin plays an important role in leukocyte adherence to microvascular endothelium and is expressed in synovial tissue from patients with rheumatoid arthritis (RA). However, the contribution of P-selectin to the initiation and chronicity of joint inflammation is not well understood. In these studies, collagen-induced arthritis (CIA) was induced in P-selectin mutant (-/-) mice to explore the role of P-selectin in the development of joint inflammation. Surprisingly, CIA onset was accelerated and severity was increased in P-selectin mutant mice, compared with wild-type mice (+/+). Increased levels of anti-type II collagen IgG were detected in both nonarthritic and arthritic P-selectin mutant mice from days 14-91. In addition, splenocytes isolated from immunized and nonimmunized P-selectin mutant mice produced significantly less IL-2 and IL-4, but significantly higher levels of IL-10 and IL-5 than splenocytes from wild-type mice. These observations show that P-selectin-mediated leukocyte rolling is not required for the development of murine CIA and that P-selectin expression exerts a controlling effect on the development of Ag-driven inflammatory joint disease, possibly by mediating the recruitment and/or trafficking of specific leukocyte subtypes into lymphoid tissue or inflammatory foci. PMID- 10453029 TI - Regulation of IL-15-stimulated TNF-alpha production by rolipram. AB - Agents that increase intracellular cAMP have been shown to reduce joint inflammation in experimental arthritis, presumably by lowering the release of proinflammatory cytokines, such as TNF-alpha. Recent studies suggest that, in joints of patients with rheumatoid arthritis, TNF-alpha release from macrophages is triggered by their interaction with IL-15-stimulated T lymphocytes. In this report, we analyze the effect of rolipram, a cAMP-specific phosphodiesterase inhibitor, on TNF-alpha production in this experimental system. Cocultures of U937 cells with IL-15-stimulated T cells, but not control T cells, resulted in increased release of TNF-alpha. Pretreatment of T cells with rolipram or cAMP analogues inhibited the IL-15-stimulated increases in proliferation, expression of cell surface molecules CD69, ICAM-1, and LFA-1, and release of TNF-alpha from macrophages. Addition of PMA to T cells dramatically increased the expression of cell surface molecules, but had little or no effect on TNF-alpha release from either T cells or from cocultures, suggesting that other surface molecules must also be involved in T cell/macrophage contact-mediated production of TNF-alpha. Addition of PMA synergistically increased the proliferation of IL-15-stimulated T cells and the secretion of TNF-alpha from IL-15-stimulated T cell/macrophage cocultures. Rolipram and 8-(4-chlorophenylthio)-cAMP (CPT-cAMP) blocked these increases. Measurement of protein kinase A (PKA) activity and the use of inhibitory cAMP analogues (RpCPT-cAMP) confirmed that rolipram worked by stimulating PKA. These data suggest that PKA-activating agents, such as rolipram, can block secretion of TNF-alpha from macrophages by inhibiting T cell activation and expression of surface molecules. PMID- 10453031 TI - Accommodated xenografts survive in the presence of anti-donor antibodies and complement that precipitate rejection of naive xenografts. AB - Hamster hearts transplanted into transiently complement-depleted and continuously cyclosporin A (CyA)-immunosuppressed rats survive long-term despite deposition of anti-donor IgM Abs and complement on the graft vascular endothelium. This phenomenon is referred to as "accommodation." The hypothesis tested here is that accommodated xenografts are resistant to IgM Abs and complement that could result in rejection of naive xenografts. After first hamster hearts had been surviving in cobra venom factor (CVF) + CyA-treated rats for 10 days, a time when the anti donor IgM Ab level was maximal and complement activity had returned to approximately 50% of pretreatment levels, naive hamster hearts or hamster hearts that had been accommodating in another rat for 14 days were transplanted into those rats carrying the surviving first graft. The naive hearts were all hyperacutely rejected. In contrast, a majority of regrafted accommodating hearts survived long-term. There was widespread Ab and activated complement deposition on the vascular endothelium of accommodating first hearts, second accommodating hearts, and rejected second naive hearts. However, only the rejected naive hearts showed extensive endothelial cell damage, myocardial necrosis, fibrin deposition, and other signs of inflammation. Accommodating first and second hearts but not rejected second naive hearts expressed high levels of the protective genes A20, heme oxygenase-1 (HO-1), bcl-2, and bcl-xL. These data demonstrate that accommodated xenografts become resistant to effects of anti-donor IgM Abs and complement that normally mediate rejection of xenografts. We hypothesize that this resistance involves expression by accommodated xenografts of protective genes. PMID- 10453032 TI - Superoxide attenuates macrophage apoptosis by NF-kappa B and AP-1 activation that promotes cyclooxygenase-2 expression. AB - Macrophages are a major source of cytokines and proinflammatory radicals such as superoxide. These mediators can be both produced and utilized by macrophages in autocrine-regulatory pathways. Therefore, we studied the potential role of oxygen radical-regulatory mechanisms in reprogramming macrophage apoptosis. Preactivation of RAW 264.7 cells with a nontoxic dose of the redox cycler 2,3 dimethoxy-1,4-naphthoquinone (5 microM) for 15 h attenuated S-nitrosoglutathione (1 mM)-initiated apoptotic cell death and averted accumulation of the tumor suppressor p53, which is indicative for macrophage apoptosis. Preactivation with superoxide promoted cyclooxygenase-2 induction that was NF-kappa B and AP-1 mediated. NF-kappa B activation was confirmed by p50/p65-heterodimer formation, I kappa B-alpha degradation, and stimulation of a NF-kappa B luciferase reporter construct. Furthermore, a NF-kappa B decoy approach abrogated cyclooxygenase-2 (Cox-2) expression as well as inducible protection. The importance of AP-1 for superoxide-mediated Cox-2 expression and cell protection was substantiated by using the extracellular signal-regulated kinase-inhibitor PD98059 and the p38 inhibitor SB203580, which blocked Cox-2 expression. In corroboration, Cox-2 expression was hindered by a dominant-negative c-jun mutant (TAM67). Protection from apoptosis was verified in human macrophages with the notion that superoxide promoted Cox-2 expression, which in turn attenuated nitric oxide-evoked caspase activation. We conclude that the sublethal generation of oxygen radicals reprograms macrophages by NF-kappa B and AP-1 activation. The resulting hyporesponsiveness reveals an attenuated apoptotic program in association with Cox-2 expression. PMID- 10453033 TI - Endothelial alpha 2,6-linked sialic acid inhibits VCAM-1-dependent adhesion under flow conditions. AB - We have previously shown that costimulation of endothelial cells with IL-1 + IL-4 markedly inhibits VCAM-1-dependent adhesion under flow conditions. We hypothesized that sialic acids on the costimulated cell surfaces may contribute to the inhibition. Northern blot analyses showed that Gal beta 1-4GlcNAc alpha 2, 6-sialyltransferase (ST6N) mRNA was up-regulated in cultured HUVEC by IL-1 or IL 4 alone, but that the expression was enhanced by costimulation, whereas the level of Gal beta 1-4GlcNAc/Gal beta 1-3GalNAc alpha2,3-sialyltransferase (ST3ON) mRNA was unchanged. Removing both alpha 2,6- and alpha 2,3-linked sialic acids from IL 1 + IL-4-costimulated HUVEC by sialidase significantly increased VCAM-1-dependent adhesion, whereas removing alpha 2,3-linked sialic acid alone had no effect; adenovirus-mediated overexpression of ST6N with costimulation almost abolished the adhesion, which was reversible by sialidase. The same treatments of IL-1 stimulated HUVEC had no effect. Lectin blotting showed that VCAM-1 is decorated with alpha 2,6- but not alpha 2,3-linked sialic acids. However, overexpression of alpha 2,6-sialyltransferase did not increase alpha 2,6-linked sialic acid on VCAM 1 but did increase alpha 2,6-linked sialic acids on other proteins that remain to be identified. These results suggest that alpha 2,6-linked sialic acids on a molecule(s) inducible by costimulation with IL-1 + IL-4 but not IL-1 alone down regulates VCAM-1-dependent adhesion under flow conditions. PMID- 10453034 TI - Eotaxin potentiates antigen-dependent basophil IL-4 production. AB - Basophils are a major source of IL-4, which is a critical factor in the generation of allergic inflammation. Eotaxin induces chemotaxis mediated through the CC chemokine receptor 3 (CCR3) present on basophils as well as eosinophils and Th2 cells, thereby promoting cell recruitment. To determine whether eotaxin has other proinflammatory activity, we examined the effect of eotaxin on basophil IL-4 expression by flow cytometry. Eotaxin alone had no effect on basophil IL-4 production, but further increased allergen-stimulated IL-4 expression. Eotaxin also enhanced IL-4 release from purified basophils 2- to 4-fold, as determined by ELISA (p < 0.01). Addition of eotaxin to cultures resulted in a 40-fold left shift in the dose response to Ag. This effect was obtained with physiologic concentrations of eotaxin (10 ng/ml), was abrogated by an Ab to the CCR3 receptor, and was noted with other chemokine ligands of CCR3. Additionally, eotaxin augmented IL-3 priming of basophil IL-4 production in a synergistic manner (p < 0.01). In contrast, no priming was observed with either IL-5 or GM CSF. These results establish a novel function for eotaxin and other chemokine ligands of CCR3: the potentiation of Ag-mediated IL-4 production in basophils, and suggest a potential nonchemotactic role for CC chemokines in the pathogenesis and amplification of inflammation. PMID- 10453035 TI - Inducible expression and regulation of the alpha 1-acid glycoprotein gene by alveolar macrophages: prostaglandin E2 and cyclic AMP act as new positive stimuli. AB - We have reported that alpha 1-acid glycoprotein (AGP) gene expression was induced in lung tissue and in alveolar type II cells during pulmonary inflammatory processes, suggesting that local production of this immunomodulatory protein might contribute to the modulation of inflammation within the alveolar space. Because AGP may also be secreted by other cell types in the alveolus, we have investigated the expression and the regulation of the AGP gene in human and rat alveolar macrophages. Spontaneous AGP secretion by alveolar macrophages was increased 4-fold in patients with interstitial lung involvement compared with that in controls. In the rat, immunoprecipitation of [35S]methionine-labeled cell lysates showed that alveolar macrophages synthesize and secrete AGP. IL-1 beta had no effect by itself, but potentiated the dexamethasone-induced increase in AGP production. RNase protection assay demonstrated that AGP mRNA, undetectable in unstimulated cells, was induced by dexamethasone. Conditioned medium from LPS stimulated macrophages as well as IL-1 beta had no effect by themselves, but potentiated the dexamethasone-induced increase in AGP mRNA levels. In addition to cytokines, PGE2 as well as dibutyryl cAMP increased AGP mRNA levels in the presence of dexamethasone. When AGP expression in other cells of the monocyte/macrophage lineage was examined, weak and no AGP production by human blood monocytes and by rat peritoneal macrophages, respectively, were observed. Our data showed that 1) AGP expression is inducible specifically in alveolar macrophages in vivo and in vitro; and 2) PGE2 and cAMP act as new positive stimuli for AGP gene expression. PMID- 10453036 TI - Signaling functions of L-selectin in neutrophils: alterations in the cytoskeleton and colocalization with CD18. AB - Ligation and clustering of L-selectin by Ab ("cross-linking") or physiologic ligands results in activation of diverse responses that favor enhanced microvascular sequestration and emigration of neutrophils. The earliest responses include a rise in intracellular calcium, enhanced tyrosine phosphorylation, and activation of extracellular signal-regulated kinases. Additionally, cross-linking of L-selectin induces sustained shape change and activation of beta2 integrins, leading to neutrophil arrest under conditions of shear flow. In this report, we examined several possible mechanisms whereby transmembrane signals from L selectin might contribute to an increase in the microvascular retention of neutrophils and enhanced efficiency of emigration. In human peripheral blood neutrophils, cross-linking of L-selectin induced alterations in cellular biophysical properties, including a decrease in cell deformability associated with F-actin assembly and redistribution, as well as enhanced adhesion of microspheres bound to beta2 integrins. L-selectin and the beta2 integrin became spatially colocalized as determined by confocal immunofluorescence microscopy and fluorescence resonance energy transfer. We conclude that intracellular signals from L-selectin may enhance the microvascular sequestration of neutrophils at sites of inflammation through a combination of cytoskeletal alterations leading to cell stiffening and an increase in adhesiveness mediated through alterations in beta2 integrins. PMID- 10453037 TI - A novel function of IL-12p40 as a chemotactic molecule for macrophages. AB - IL-12p70 plays a pivotal role in regulating the Th1/Th2 balance in the initial stage of immune responses. In contrast, IL-12p40, which is produced excess over IL-12p70, has been known to down-regulate IL-12p70-mediated responses by acting as an antagonist. To investigate in vivo function of IL-12p40, RH7777 rat hepatoma cells were engineered to inducibly express mouse IL-12p40 under the tight control of doxycycline (dox). In the absence of dox, s.c. injection of these cells into syngeneic rat was shown to generate tumors. However, the induction of IL-12p40 by dox was sufficient for inhibiting tumor formation, as well as for tumor regression. Immunohistochemical analysis showed that macrophages, but not CD4+ T, CD8+ T, and NK cells, were predominantly recruited into tumor sites as early as 3 days after IL-12p40 induction. These results were further supported by the observation that IL-12p40, but not C-terminal deletion mutants by more than 5 amino acids, was able to chemoattract peritoneal macrophages in vitro, suggesting that IL-12p40, when produced in a large excess over IL-12p70 in vivo, can initially amplify the immune responses against tumors by directly recruiting macrophages. Our findings indicate that IL-12p40 may function as an effector molecule as well as an antagonist of IL-12p70. PMID- 10453038 TI - Macrolide antibiotics protect against immune complex-induced lung injury in rats: role of nitric oxide from alveolar macrophages. AB - Macrolide antibiotics have unique immunomodulatory actions apart from antimicrobial properties. We studied the effects of macrolides on IgG immune complex (IgG-ICx)-induced lung injury in rats in vivo and in vitro. Intrapulmonary deposition of IgG-ICx produced a time-dependent increase in the concentration of NO in exhaled air. There were corresponding increases in the number of neutrophils accumulated into alveolar spaces, and lung wet-to-dry weight ratio. All of these changes were inhibited by pretreatment with erythromycin or josamycin, but not by amoxicillin or cephaclor. Incubation of cultured pulmonary alveolar macrophages caused up-regulation of NO production and expression of inducible NO synthase mRNA, an effect that was dose dependently inhibited by erythromycin, roxithromycin, or josamycin. The macrolides also reduced IgG-ICx-induced release of IL-1beta and TNF-alpha, but did not alter the release of NO induced by exogenously added IL-1beta and TNF-alpha. These results suggest that macrolide antibiotics specifically inhibit immune complex-induced lung injury presumably by inhibiting cytokine release and the resultant down regulation of inducible NO synthase gene expression and NO production by rat pulmonary alveolar macrophages. PMID- 10453039 TI - Identification of genes controlling collagen-induced arthritis in mice: striking homology with susceptibility loci previously identified in the rat. AB - The susceptibility to collagen-induced arthritis in the highly susceptible DBA/1 mouse has earlier been shown to be partly controlled by the MHC class II gene Aq. To identify susceptibility loci outside of MHC, we have made crosses between DBA/1 and the less susceptible B10.Q strain, both expressing the MHC class II gene Aq. Analysis of 224 F2 intercross mice with 170 microsatellite markers in a genome-wide scan suggested 4 quantitative trait loci controlling arthritis susceptibility located on chromosomes 6, 7, 8, and 10. The locus on chromosome 6 (Cia6), which was associated with arthritis onset, yielded a logarithm of odds score of 4.7 in the F2 intercross experiment and was reproduced in serial backcross experiments. Surprisingly, the DBA/1 allele had a recessive effect leading to a delay in arthritis onset. The suggestive loci on chromosomes 7 and 10 were associated with arthritis severity rather than onset, and another suggestive locus on chromosome 8 was most closely associated with arthritis incidence. The loci on chromosomes 7, 8, and 10 all appeared to contain disease promoting alleles derived from the DBA/1 strain. Interestingly, most of the identified loci were situated in chromosomal regions that are homologous to regions in the rat genome containing susceptibility genes for arthritis; the mouse Cia6 locus is homologous with the rat Cia3, Pia5, Pia2, and Aia3; the locus on chromosome 7 (Cia7) is homologous with the rat Cia2; and the locus on chromosome 10 (Cia8) is homologous with the rat Cia4. PMID- 10453040 TI - GDKV-induced antiphospholipid antibodies enhance thrombosis and activate endothelial cells in vivo and in vitro. AB - Antiphospholipid (aPL) Abs are associated with thrombosis, pregnancy loss, and thrombocytopenia in patients with systemic lupus erythematosus or primary antiphospholipid syndrome (APS). beta2-Glycoprotein I (beta2GPI), a phospholipid binding serum protein, is involved in aPL binding to phospholipids. aPL can be generated in mice by immunization with beta2GPI, and these Abs are thrombogenic and cause pregnancy loss in mice. The objective of this study is to determine whether aPL induced by immunization with the phospholipid-binding site of beta2GPI are thrombogenic and whether they activate endothelial cells (EC) in vivo and in vitro. Murine monoclonal aPL were generated from spleen cells of a mouse immunized with GDKV, a synthetic 15-aa peptide spanning Gly274-Cys288 in the fifth domain of human beta2GPI, which represents the phospholipid-binding site of beta2GPI. The Abs generated had aPL and anti-beta2GPI activities. The effect of these Abs on thrombus formation and on EC activation in vivo was determined using a mouse model of thrombosis and microcirculation that enables examination of the adhesion of leukocyte to EC as an indication of EC activation as well as adhesion molecule expression using in vitro ELISA analysis. Mice injected with this monoclonal aPL showed a significant increase in leukocyte sticking and also produced larger thrombi that persisted longer. Exposure to GDKV induced aPL for 4 h significantly increased surface Ag expression of E-selectin, ICAM-1, and VCAM-1. These data indicate that aPL induced by immunization with the phospholipid binding site of beta2GPI are thrombogenic and activate endothelial cells. PMID- 10453041 TI - Identification of five MAGE-A1 epitopes recognized by cytolytic T lymphocytes obtained by in vitro stimulation with dendritic cells transduced with MAGE-A1. AB - MAGE genes are expressed by many human tumors of different histological types but not by normal cells, except for male germline cells. The Ags encoded by MAGE genes and recognized by T cells are therefore strictly tumor-specific. Clinical trials involving therapeutic vaccination of cancer patients with MAGE antigenic peptides or proteins are in progress. To increase the range of patients eligible for therapy with peptides, it is important to identify additional MAGE epitopes recognized by CTL. Candidate peptides known to bind to a given HLA have been used to stimulate T lymphocytes in vitro. In some instances, CTL clones directed against these synthetic peptides have been obtained, but these clones often failed to recognize tumor cells expressing the relevant gene. Therefore, we designed a method to identify CTL epitopes that selects naturally processed peptides. Monocyte-derived dendritic cells infected with a recombinant canarypoxvirus (ALVAC) containing the entire MAGE-A1 gene were used to stimulate CD8+ T lymphocytes from the blood of individuals without cancer. Responder cell microcultures that specifically lysed autologous cells expressing MAGE-A1 were cloned using autologous stimulator cells either transduced with a retrovirus coding for MAGE-A1 or infected with recombinant Yersinia-MAGE-A1 bacteria. The CTL clones were tested for their ability to lyse autologous cells loaded with each of a set of overlapping MAGE-A1 peptides. This strategy led to the identification of five new MAGE-A1 epitopes recognized by CTL clones on HLA-A3, A28, -B53, -Cw2, and -Cw3 molecules. All of these CTL clones recognized target cells expressing gene MAGE-A1. PMID- 10453042 TI - Experimental autoimmune encephalomyelitis in NF-kappa B-deficient mice:roles of NF-kappa B in the activation and differentiation of autoreactive T cells. AB - Experimental autoimmune encephalomyelitis (EAE) is an inflammatory disease of the CNS, which has long been used as an animal model for human multiple sclerosis. Development of autoimmune disease requires coordinated expression of a number of genes that are involved in the activation and effector functions of inflammatory cells. These include genes that encode costimulatory molecules, cytokines, chemokines, and adhesion molecules. Activation of these genes is regulated at the transcriptional level by several families of transcription factors. One of these is the NF-kappa B family, which is present in a variety of cell types and becomes highly activated at sites of inflammation. To test the roles of NF-kappa B in the development of autoimmune diseases, we studied EAE in mice deficient in one of the NF-kappa B isoforms, i.e., NF-kappa B1 (p50). We found that NF-kappa B1 deficient mice were significantly resistant to EAE induced by myelin oligodendrocyte glycoprotein. The resistance was primarily evidenced by a decrease in disease incidence, clinical score, and the degree of CNS inflammation. Furthermore, we established that the resistance to EAE in NF-kappa B1-deficient mice was associated with a deficiency of myelin oligodendrocyte glycoprotein-specific T cells to differentiate into either Th1- or Th2-type effector cells in vivo. These results strongly suggest that NF-kappa B1 plays crucial roles in the activation and differentiation of autoreactive T cells in vivo and that blocking NF-kappa B function can be an effective means to prevent autoimmune encephalomyelitis. PMID- 10453043 TI - Blocking antibodies induced by specific allergy vaccination prevent the activation of CD4+ T cells by inhibiting serum-IgE-facilitated allergen presentation. AB - Allergen-specific CD4+ T lymphocytes are activated at extremely low allergen concentrations in vivo as a result of serum-facilitated allergen presentation (S FAP). It is not clear at present if specific allergy vaccination (SAV) has an effect on this mechanism. Here we show that birch allergen-specific serum-IgE facilitates the presentation of Bet v 1, the major birch pollen allergen, to Bet v 1-specific CD4+ T lymphocytes by a factor of >100. This process is CD23 mediated, could be detected in sera from the majority of birch-allergic patients, and was clearly dose dependent. S-FAP of Bet v 1 was inhibited in patients undergoing long-term birch SAV, but not by sera from patients undergoing grass SAV, indicating that birch-specific Abs are involved. This resulted in decreased proliferation and IL-4, IL-5, IL-10, and IFN-gamma production of Bet v 1-specific T cells. The inhibition was already noted after 3-9 mo of SAV and could not be solely explained by increased serum levels of birch-specific IgG4. When IgG- and IgA/IgM-containing fractions of long-term SAV sera were used to inhibit S-FAP, only IgG-containing fractions were shown to inhibit S-FAP. These results indicate that blocking IgG Abs induced by SAV inhibits the occurrence of S-FAP at very low allergen concentrations, resulting in significantly higher allergen threshold levels to obtain T cell proliferation and cytokine production and thus allergen induced late-phase responses. PMID- 10453044 TI - HIV-1 Tat induces monocyte chemoattractant protein-1-mediated monocyte transmigration across a model of the human blood-brain barrier and up-regulates CCR5 expression on human monocytes. AB - AIDS dementia is characterized by neuronal loss in association with synaptic damage. A central predictor for clinical onset of these symptoms is the infiltration of monocytes and macrophages into CNS parenchyma. Chronic HIV-1 infection of monocytes also allows these cells to serve as reservoirs for persistent viral infection. Using a coculture of endothelial cells and astrocytes that models several aspects of the human blood-brain barrier, we examined the mechanism whereby the HIV-derived factor Tat may facilitate monocyte transmigration. We demonstrate that treatment of cocultures on the astrocyte side with HIV-1 Tat induced significant monocyte chemoattractant protein (MCP)-1 protein. Astrocytes, but not endothelial cells, were the source of this MCP-1 expression. Supernatants from Tat-treated cocultures induced significant monocyte transmigration, which was detected by 2.5 h after the addition of PBMC. Pretreatment of the supernatants from Tat-stimulated cocultures with an Ab to MCP 1 completely blocked monocyte transmigration. Flow cytometric analysis of Tat stimulated PBMC demonstrated that Tat up-regulated expression of the chemokine receptor, CCR5, on monocytes in a time-dependent manner. Taken together, our data indicate that HIV-1 Tat may facilitate the recruitment of monocytes into the CNS by inducing MCP-1 expression in astrocytes. These recruited monocytes may contribute to the pathogenesis of HIV-1-associated AIDS encephalitis and dementia. PMID- 10453046 TI - Introduction to the journal of neurobiology special issue: glia and steroids PMID- 10453045 TI - Pancreas-infiltrating Th1 cells and diabetes develop in IL-12-deficient nonobese diabetic mice. AB - IL-12 and IL-12 antagonist administration to nonobese diabetic (NOD) mice accelerates and prevents insulin-dependent diabetes mellitus (IDDM), respectively. To further define the role of endogenous IL-12 in the development of diabetogenic Th1 cells, IL-12-deficient NOD mice were generated and analyzed. Th1 responses to exogenous Ags were reduced by approximately 80% in draining lymph nodes of these mice, and addition of IL-12, but not IL-18, restored Th1 development in vitro, indicating a nonredundant role of IL-12. Moreover, spontaneous Th1 responses to a self Ag, the tyrosine phosphatase-like IA-2, were undetectable in lymphoid organs from IL-12-deficient, in contrast to wild-type, NOD mice. Nevertheless, wild-type and IL-12-deficient NOD mice developed similar insulitis and IDDM. Both in wild-type and IL-12-deficient NOD mice, approximately 20% of pancreas-infiltrating CD4+ T cells produced IFN-gamma, whereas very few produced IL-10 or IL-4, indicating that IDDM was associated with a type 1 T cell infiltrate in the target organ. T cell recruitment in the pancreas seemed favored in IL-12-deficient NOD mice, as revealed by increased P-selectin ligand expression on pancreas-infiltrating T cells, and this could, at least in part, compensate for the defective Th1 cell pool recruitable from peripheral lymphoid organs. Residual Th1 cells could also accumulate in the pancreas of IL-12 deficient NOD mice because Th2 cells were not induced, in contrast to wild-type NOD mice treated with an IL-12 antagonist. Thus, a regulatory pathway seems necessary to counteract the pathogenic Th1 cells that develop in the absence of IL-12 in a spontaneous chronic progressive autoimmune disease under polygenic control, such as IDDM. PMID- 10453047 TI - Glia as mediators of steroid hormone action on the nervous system: An overview. AB - This special issue on steroids and glia represents the intersection of two emerging themes in the neurosciences: (a) Glia actively modulate and participate in brain function throughout life, and (b) glia are sensitive to steroid hormones. This overview begins by reviewing some of the basic principles of steroid hormone action on the brain and introducing the various glia that inhabit the peripheral and central nervous system. A prominent theme among the articles that follow is that glia may be direct targets for steroid hormones since they possess steroid receptors and the promoter region of glial-specific genes such as glutamine synthetase contain hormone-responsive elements. The articles in this special issue discuss evidence that glia may mediate steroid action on the nervous system in the context of (a) steroid metabolism, which may control the hormonal microenvironment of neurons both in the normal and injured brain; (b) brain development including sexual differentiation; (c) synaptic plasticity which may underlie the cyclic release of luteinizing hormone releasing hormone in the female rodent brain; (d) neural repair and aging; and (e) brain immune function. Another theme among these articles is that glia influence neurons via specific secreted and cell-surface molecules, and that steroids affect this mode of communication by altering the level of glial production of these signaling molecules and/or the sensitivity of neurons to such signals. PMID- 10453049 TI - Estrogen receptor immunoreactivity in Schwann-like brain macroglia. AB - Olfactory ensheathing cells, tanycytes, pituicytes, pineal glia, retinal Muller cells, and Bergmann glia of normal male rats express concomitantly estrogen receptor, low-affinity neurotrophin receptor, antigen O4, and GFAP, markers characteristic of nonmyelinating Schwann cells. These cells were able to survive and proliferate when cultured from adult tissue, promoted neurite outgrowth, and could guide and ensheath growing neurites. We called this distinct group of growth-promoting central nervous system (CNS) macroglia aldynoglia (Greek: to make grow). Its proliferative and growth-promoting properties seem to be retained during the whole lifetime of the organism in those CNS loci where normal function depends on continuous axon renewal. Aldynoglia plasticity seems totally or partially lost with age where and when it is no longer critical, as in the case of adult cortical and spinal cord radial glia. The concomitant expression of estrogen receptor and low-affinity neurotrophin receptor may promote Schwann-like plasticity of glial cells. PMID- 10453048 TI - Immunoreactivity for intracellular androgen receptors in identified subpopulations of neurons, astrocytes and oligodendrocytes in primate prefrontal cortex. AB - Sex differences in and hormone malleability of a variety of cognitive and mnemonic functions suggest that the association cortices in human and nonhuman primates are targets of gonadal hormone stimulation. One mechanism involved in this stimulation may be genomic actions mediated by intracellular androgen receptors. To identify potential cellular targets of this influence, single- and double-labeling immunohistochemical methods were used to precisely localize androgen receptor proteins in the prefrontal association cortex of adult rhesus monkeys. In both the dorsolateral and orbitofrontal regions, receptor antibodies labeled substantial populations of small intensely immunoreactive nuclei, as well as much larger and less strongly immunoreactive nuclei in all major cellular layers and/or in underlying white matter. Double-labeling studies revealed that large and small immunolabeled nuclei were further distinguished by colocalization with different classes of cell-specific markers. Whereas the large, pale receptor immunoreactive nuclei colocalized with immunomarkers for neurons, the small, strongly immunoreactive nuclei colocalized exclusively with glial markers. Among androgen receptor-immunoreactive glia, a majority were immunoreactive for astrocyte markers, with smaller numbers of nuclei colocalized with oligodendrocyte markers; immunolabels for microglia failed to colocalize with androgen receptor immunoreactivity. This discovery of an unexpectedly large population of androgen receptor bearing glia suggests that direct functional interactions between endocrine signaling pathways and glial cells such as those coming into view in studies in subcortical and allocortical structures may also take place in the cerebral cortex and contribute to gonadal hormone stimulation of cortical processing of cognitive information. PMID- 10453050 TI - Steroid metabolism and effects in central and peripheral glial cells. AB - Hormonal steroids participate in the control of a large number of functions of the central nervous system (CNS); recent data show that they may also intervene at the level of the peripheral nervous system (PNS). Both the CNS and the PNS metabolize endogenous as well as exogenous steroids; one of the major enzymatic system is represented by the 5alpha-reductase-3alpha-hydroxysteroid complex. This is a versatile system, since every steroid possessing the delta 4-3keto configuration (e.g., testosterone, progesterone, deoxycorticosterone) may be a substrate. High levels of 5alpha-reductase are found in the white matter of the CNS and in purified myelin. The observation that, in addition to neurons, glia may be a target for steroid action is an important recent finding. The effects of progesterone, testosterone, corticoids, and their respective 5alpha and 3alpha 5alpha derivatives on the expression of glial genes are presented and discussed. It has also been found that progesterone and/or its 5alpha-reduced metabolites increase the mRNA for the two major proteins of peripheral myelin, the glycoprotein Po and the peripheral myelin protein 22, in the sciatic nerve of normal and aged animals and in Schwann cells. The hypothesis has been put forward that glycoprotein Po might be under the control of progestagens acting mainly via the progesterone receptor, and that peripheral myelin protein 22 might be controlled via an interaction of steroids with the gamma-aminobutyric acid (GABA)ergic system. It is known that tetrahydroprogesterone, the 3alpha-5alpha reduced metabolite of progesterone, interacts with the GABA(A) receptor. Our recent data show that several subunits of this receptor are present in sciatic nerve as well as in Schwann cells that reside in this nerve. These data open multiple possibilities for new therapeutic approaches to demyelinating diseases. PMID- 10453051 TI - Estrogen and microglia: A regulatory system that affects the brain. AB - Sex hormones are involved in the physiological regulation of several aspects of behavior and neuroendocrine events. It has been accepted that such effects are mediated directly by steroid actions on neurons; however, new studies have shown that the glial cells are also affected by gonadal steroids. The microglia are one specialized brain glial cell type, which is a target for estrogen actions. In fact, we believe that many of the immune and nonimmune regulatory functions of microglia in the brain are influenced directly by estrogen via expression and secretion of cytokines, and growth factors by the microglia. The present review details only a section of the known aspects of microglial function, focusing mainly on nonimmune regulatory actions in the brain and their functional relationship with sex hormones. Moreover, we present evidence for the presence of estrogen receptor-beta (ERbeta) in rat microglial cells. PMID- 10453052 TI - Oligodendrocyte development and thyroid hormone. AB - Thyroid hormone plays an important role in brain development and is essential to ensure a normal myelination. The effects of thyroid hormone are mediated by nuclear thyroid hormone receptors, which act as ligand-regulated transcription factors. There are several isoforms encoded by two genes, alpha and beta. Developmental studies have shown that alpha isoforms are widely expressed in the fetal brain, while beta isoforms expression is more restricted with a dramatic increase that begins at birth in the rat. Remarkably, receptor number reaches maximal levels by postnatal day 10, when serum thyroid hormone levels also peak and myelination is the most prominent event in the developing rat brain. Likewise, oligodendrocyte precursor cells express alpha isoforms and expression of the beta isoforms is confined to the differentiated oligodendrocytes, suggesting that these isoforms might mediate different thyroid hormone effects in the oligodendrocyte lineage. Thyroid hormone acts at multiple steps in the development of oligodendrocytes: (a) Early in development, it can function as an instructive signal for the generation of oligodendrocytes and enhance the proliferation of the committed preprecursor oligodendrocyte cells. (b) Thyroid hormone regulates the number of oligodendrocyte generated by directly promoting their differentiation. Since oligodendrocytes are produced in vitro after the same period in culture regardless of whether thyroid hormone was added to the cultures, it has been suggested that thyroid hormone is required for neither the timing nor the generation of oligodendrocytes, but is necessary to achieve adequate oligodendrocyte numbers. (c) Finally, thyroid hormone increases morphological and functional maturation of postmitoitic oligodendrocytes by stimulation of the expression of various myelin genes. PMID- 10453053 TI - Glucocorticoid control of glial gene expression. AB - The glucocorticoid signaling pathway is responsive to a considerable number of internal and external signals and can therefore establish diverse patterns of gene expression. A glial-specific pattern, for example, is shown by the glucocorticoid-inducible gene glutamine synthetase. The enzyme is expressed at a particularly high level in glial cells, where it catalyzes the recycling of the neurotransmitter glutamate, and at a low level in most other cells, for housekeeping duties. Glial specificity of glutamine synthetase induction is achieved by the use of positive and negative regulatory elements, a glucocorticoid response element and a neural restrictive silencer element. Though not glial specific by themselves, these elements may establish a glial-specific pattern of expression through their mutual activity and their combined effect. The inductive activity of glucocorticoids is markedly repressed by the c-Jun protein, which is expressed at relatively high levels in proliferating glial cells. The signaling pathway of c-Jun is activated by the disruption of glia neuron cell contacts, by transformation with v-src, and in proliferating retinal cells of early embryonic ages. The c-Jun protein inhibits the transcriptional activity of the glucocorticoid receptor and thus represses glutamine synthetase expression. This repressive mechanism might also affect the ability of glial cells to cope with glutamate neurotoxicity in injured tissues. PMID- 10453054 TI - Glial-neuronal interactions in the neuroendocrine control of mammalian puberty: facilitatory effects of gonadal steroids. AB - It is now clear that astroglial cells actively contribute to both the generation and flow of information within the central nervous system. In the hypothalamus, astrocytes regulate the secretory activity of neuroendocrine neurons. A small subset of these neurons secrete luteinizing hormone-releasing hormone (LHRH), a neuropeptide essential for sexual development and adult reproductive function. Astrocytes stimulate LHRH secretion via cell-cell signaling mechanisms involving growth factors recognized by receptors with either serine/threonine or tyrosine kinase activity. Two members of the epidermal growth factor (EGF) family and their respective tyrosine kinase receptors appear to play key roles in this regulatory process. Transforming growth factor-alpha (TGFalpha) and its distant congeners, the neuregulins (NRGs), are produced in hypothalamic astrocytes. They stimulate LHRH secretion indirectly, via activation of erbB-1/erbB-2 and erbB 4/erbB-2 receptor complexes also located on astrocytes. Activation of these receptors leads to release of prostaglandin E(2) (PGE(2)), which then binds to specific receptors on LHRH neurons to elicit LHRH secretion. Gonadal steroids facilitate this glia-to-neuron communication process by acting at three different steps along the signaling pathway. They (a) increase astrocytic gene expression of at least one of the EGF-related ligands (TGFalpha), (b) increase expression of at least two of the receptors (erbB-4 and erbB-2), and (c) enhance the LHRH response to PGE(2) by up-regulating in LHRH neurons the expression of specific PGE(2) receptor isoforms. Focal overexpression of TGFalpha in either the median eminence or preoptic area of the hypothalamus accelerates puberty. Conversely, blockade of either TGFalpha or NRG hypothalamic actions delays the process. Thus, both TGFalpha and NRGs appear to be physiological components of the central neuroendocrine mechanism controlling the initiation of female puberty. By facilitating growth factor signaling pathways in the hypothalamus, ovarian steroids accelerate the pace and progression of the pubertal process. PMID- 10453055 TI - Role of thyroid hormones and their receptors in peripheral nerve regeneration. AB - After peripheral nerve injury in adult mammals, reestablishment of functional connections depends on several parameters including neurotrophic factors, the extracellular matrix, and hormones. However, little is known about the contribution of hormones to peripheral nerve regeneration. Thyroid hormones, which are required for the development and maturation of the central nervous system, are also important for the development of peripheral nerves. The action of triiodothyronine (T3) on responsive cells is mediated through nuclear thyroid hormone receptors (TRs) which modulate the expression of specific genes in target cells. Thus, to study the effect of T3, it is first necessary to know whether the target tissues possess TRs. The fact that sciatic nerve cells possess functional TRs suggests that these cells can respond to T3 and, as a consequence, that thyroid hormone may be involved in peripheral nerve regeneration. The silicone nerve guide model provides an excellent system to study the action of local administration of T3. Evidence from such studies demonstrate that animals treated locally with T3 at the level of transection have more complete regeneration of sciatic nerve and better functional recovery. Among the possible regulatory mechanisms by which T3 enhances peripheral nerve regeneration is rapid action on both axotomized neurons and Schwann cells which, in turn, produce a lasting and stimulatory effect on peripheral nerve regeneration. It is probable that T3 up- or down-regulates gene expression of one or more growth factors, extracellular matrix, or cell adhesion molecules, all of which stimulate peripheral nerve regeneration. This could explain the greater effect of T3 on nerve regeneration compared with the effect of any one growth factor or adhesion molecule. PMID- 10453056 TI - Androgenic regulation of the central glia response following nerve damage. AB - Current research on the effects of gonadal steroids on the brain and spinal cord indicates that these agents have profound trophic effects on many aspects of neuronal functioning, including cell survival, growth and metabolism, elaboration of processes, synaptogenesis, and neurotransmission (Jones et al., 1985; Luine, 1985; Nordeen et al., 1985; Matsumoto et al., 1988a,b; Gould et al., 1990). Since many of the aspects of normal neuronal functioning altered by gonadal steroids are affected by injury to the nervous system, we initiated a series of experiments designed to exploit the trophic capabilities of steroids as therapeutic agents in neuronal injury and repair (Kujawa et al., 1989, 1991; Kujawa and Jones, 1990). Three steroid-sensitive model systems were used for these studies: the hamster facial motoneuron, the rat sciatic motoneuron, and the hamster rubrospinal motoneuron. The results of our initial series of experiments suggest that androgens, and possibly estrogens, act either directly or indirectly on the injured motoneuron and enhance elements of the neuronal reparative response that are critical to successful recovery of function. Recently, we discovered that gonadal steroids may also modulate the central glia response to nerve damage. In this review, a summary of our data identifying a therapeutic role for androgens in enhancing the reparative response of motoneurons to injury is presented. This is followed by a discussion of the effects of androgens on the glial response to injury. PMID- 10453057 TI - Role of astroglia in estrogen regulation of synaptic plasticity and brain repair. AB - Astroglia are targets for estrogen and testosterone and are apparently involved in the action of sex steroids on the brain. Sex hormones induce changes in the expression of glial fibrillary acidic protein, the growth of astrocytic processes, and the degree of apposition of astroglial processes to neuronal membranes in the rat hypothalamus. These changes are linked to modifications in the number of synaptic inputs to hypothalamic neurons. These findings suggest that astrocytes may participate in the genesis of androgen-induced sex differences in synaptic connectivity and in estrogen-induced synaptic plasticity in the adult brain. Astrocytes and tanycytes may also participate in the cellular effects of sex steroids by releasing neuroactive substances and by regulating the local accumulation of specific growth factors, such as insulin-like growth factor I, that are involved in estrogen-induced synaptic plasticity and estrogen mediated neuroendocrine control. Astroglia may also be involved in regenerative and neuroprotective effects of sex steroids, since astroglia formation after brain injury or after peripheral nerve axotomy is regulated by sex hormones. Furthermore, the expression of aromatase, the enzyme that produces estrogen, is induced de novo in astrocytes in lesioned brain areas of adult male and female rodents. Since astroglia do not express aromatase under normal circumstances, the induction of this enzyme may be part of the program of glial activation to cope with the new conditions of the neural tissue after injury. Given the neuroprotective and growth-promoting effects of estrogen after injury, the local production of this steroid may be a relevant component of the reparative process. PMID- 10453058 TI - Glial responses to steroids as markers of brain aging. AB - Glia mediate neuroendocrine and neuroimmune functions that are altered during the process of normal aging. The biological functions of glia are also important in synaptic remodeling and the loss of synaptic connections that occur during aging. These functions are carried out by changes in glia, including changes in shape, interactions with neurons and other glia, and gene expression. The predominant change that occurs in glia during aging is glial activation, which can progress to reactive gliosis in response to neurodegeneration. More markers are needed to distinguish normal and reactive glia. During aging, astrocytes hypertrophy and exhibit signs of metabolic activation, and astrocytic processes surround neurons. Microglia also become activated and subsets of activated microglial increase in number and may enter the phagocytic or reactive stage. Glial markers of brain aging and glial activation include glial fibrillary acidic protein (GFAP) and transforming growth factor (TGF)-beta1, which are increased in astrocytes and microglia, respectively. Steroids regulate the interactions between glia and neurons and glial gene expression, including GFAP and TGF-beta1. Therefore, changes in these parameters during aging may be due to altered steroid regulation. In general, the effects of steroids oppose the effects of aging. Recent data indicate that steroid treatment can decrease the expression of GFAP in the aged brain, yet GFAP is resistant to down-regulation by endogenous glucocorticoids. Cellular and molecular markers of glial activation are being used to determine how changes in neuroendocrine and neuroimmune regulation contribute to repair and functional recovery that may reverse synaptic loss and cognitive impairment during aging. PMID- 10453059 TI - Steroid-induced developmental plasticity in hypothalamic astrocytes: implications for synaptic patterning. AB - We have previously demonstrated that astrocytes in the developing arcuate nucleus of the rat hypothalamus exhibit a sexually dimorphic morphology as a result of differential exposure to gonadal steroids. Testosterone via its aromatized byproduct, estrogen, induces arcuate astrocytes to undergo differentiation during the first few days of life. These differentiated astrocytes exhibit a stellate morphology. Coincident with the steroid-induced increase in astrocyte differentiation is a reduction of dendritic spines on arcuate neurons. As a result, the arcuate nucleus of males has fewer axodendritic spine synapses than females and this dimorphism is retained throughout life. In the immediately adjacent ventromedial nucleus, neonatal astrocytes are immature and unresponsive to steroids. Neurons in this region show no change in dendritic spines in the first few days of life but do exhibit increased dendritic branching as a result of testosterone exposure. These findings illustrate the importance of distinct populations of astrocytes in restricted brain regions and their potential importance to the establishment of regionally specific synaptic patterning. Conflicting reports leave the site of steroid-mediated astrocyte responsiveness in the arcuate nucleus unresolved: Are gonadal steroids acting directly on astrocytes or are steroid-concentrating neurons mediating astrocytic responsiveness? In this review, we discuss the current understanding of astrocyte neuron interactions and the possible mechanisms for steroid-mediated, astrocyte directed synaptic patterning in the developing hypothalamus. PMID- 10453060 TI - Comparison between personal exposure to 60 Hz magnetic fields and stationary home measurements for people living near and away from a 735 kV power line. AB - This study compares stationary home measurements with a personal exposure monitor of 60 Hz magnetic fields in a group of 18 people living near a 735 kV line and 17 people living far away from the line. Most of them were white collar workers who worked during the day. They wore a personal Positron meter for 24 h, while a similar meter was left in their home, away from any appliances. For people living away from the line, the impact of residential activities appeared rather weak when considering the average intensity of the field during the awake period (at home): 0.22 microT for personal exposure versus 0.18 microT for stationary measurements (P = 0.09). The impact of residential activities during the awake period was more detectable when using the percentage of time with exposure above 0.78 microT: median 0.4 for personal vs. 0.0 for stationary measurements (P =.01). The temporal variability of the exposure during the awake period was also significantly higher for personal exposure than for stationary measurements. For people living near the line, the intensity of the magnetic field from the line dominated the personal exposure when considering the mean of measurements and the percentage of time above a threshold. However, the temporal variability was greater for the personal exposure during the awake period. Although limited due to its small sample size, the present study seems to demonstrate the usefulness of considering different indexes of exposure when assessing residential exposure to 60 Hz magnetic fields. PMID- 10453061 TI - Alterations in ornithine decarboxylase activity in the rat mammary gland after different periods of 50 Hz magnetic field exposure. AB - In a series of experiments with the chemical carcinogen DMBA (7, 12 dimethyl[a]anthracene), we recently found that exposure of female Sprague-Dawley rats in 50 Hz magnetic fields (MF) in the microtesla range significantly facilitates the development and growth of mammary tumors. One possible explanation for this finding would be enhanced proliferation of breast epithelial stem cells by MF exposure, thereby increasing the sensitivity of these cells to chemical carcinogens. In line with this possibility, we previously determined that 50 Hz, 50 microT MF exposure induces increases in ornithine decarboxylase (ODC), i.e., a key enzyme in cell proliferation, in the mammary gland of female Sprague-Dawley rats. In the present study, we examined the time course of this effect, by using different periods of exposure to a 50 Hz, 100 microT MF. Furthermore, we determined ODC in different mammary complexes of the rat mammary gland to evaluate whether differences in response to MF exist over the anterior posterior extension of this organ. Exposure of young female Sprague-Dawley rats induced marked increases in ODC in the mammary gland that were similar to ODC increases seen in "positive control" experiments with the tumor promoter 12-O tetradecanoylphorbol-13-acetate (TPA). However, this effect of MF critically depended on the duration of MF exposure, with no effect, or at least no consistent effect, for short (<1 week) or long (8 weeks and above) exposure periods, but a robust and reproducible enhancing effect on ODC activity after 2 weeks of exposure. Furthermore, we found that the effect of MF exposure depends on the part of the mammary complexes examined, the cranial thoracic (or cervical) complexes being particularly sensitive to ODC alterations in response to MF. This is in line with recent DMBA experiments of our group in which MF-induced increases in tumor development and growth were predominantly seen in this large cranial/cervical part of the mammary gland. The most likely explanation for the observed ODC changes after MF exposure is the "melatonin hypothesis," although other cellular and molecular effects of MF might be involved as well. PMID- 10453062 TI - Induction of stress proteins by electromagnetic fields in cultured HL-60 cells. AB - HL-60 cells in culture were exposed for 2 h to a sinusoidal 0.1 or 1 mT (1 or 10 Gauss) magnetic field at 60 Hz and pulse labeled after exposure with radioactive isotopes by incubation by using either [(35)S]methionine, [(3)H]leucine, or [(33)P]phosphate. The radioactive labels were incorporated into cellular proteins through synthesis or phosphorylation. Proteins were extracted from electrostatically sorted nuclei, and the heat shock/stress proteins (sp) were analyzed for synthesis and phosphorylation by two-dimensional polyacrylamide gel electrophoresis. In the control cultures (no exposure to the magnetic field), sp 72c (cognate form) was faintly observed. A 0.1 mT exposure did not show sp metabolism to be different from that of the controls; however, after a 1 mT exposure of the HL-60 cells, sp 70i (inducible form) was synthesized ([(35)S]methionine incorporation). Sp 90 was not synthesized at either field level, but was phosphorylated ([(33)P]phosphate incorporation) in the 1 mT exposure. Sp 27 (isoforms a and b) was induced after a 1 mT exposure as reflected by labeling with [(3)H]leucine. These sps were not detected after a 0.1 mT exposure. After a 1 mT exposure and labeling with [(33)P], sp 27 isoforms b and c were phosphorylated whereas isoform 'a' was not observed. Sps 70i, 72c, and 90 were identified by commercial sp antibodies. Likewise, polypeptides a, b, and c were verified as sp 27 isoforms by Western blotting. Statistical evaluation of sp areas and densities, determined from fluorographs by Western-blot analysis, revealed a significant increase in sps 90 and 27a after a 1 mT magnetic field exposure. The 1 mT magnetic field interacts at the cellular level to induce a variety of sp species. Bioelectromagnetics 20:347-357, 1999. Published 1999 Wiley Liss, Inc. PMID- 10453063 TI - Macro- and trace element concentrations in blood plasma and cerebrospinal fluid of dairy cows exposed to electric and magnetic fields. AB - Eight multiparous, nonlactating pregnant Holstein cows (at 198 +/- 35 days of gestation and weighing 608 + 24 kg) and seven nonlactating nonpregnant ovariectomized heifers (weighing 370 + 29 kg) were confined to wooden metabolism crates in an electric and magnetic field chamber. Subarachnoidal catheters were inserted before the activation of the electric and magnetic fields. For 30 days, cows and heifers were continuously exposed in separate trials to electric and magnetic fields (60 Hz, 10 kV/m, and 30 microT). Blood plasma and cerebrospinal fluid samples were collected for 3 consecutive days before the exposure period, the last 3 days of the exposure period, and for 3 days starting 5 days after the exposure period. Concentrations of Ca, Mg, Cu, Zn, Fe, Mn, Na, P, and K in blood plasma and cerebrospinal fluid were determined. Exposure to electric and magnetic fields resulted in decreased concentrations of Mg in blood plasma and in increased concentrations of Ca and P and decreased concentrations of Fe and Mn in cerebrospinal fluid. PMID- 10453064 TI - Analytic SAR computation in a multilayer elliptic cylinder for bioelectromagnetic applications. AB - The specific absorption rate (SAR) is usually considered as the basic quantity to derive the reference levels for the exposure of workers and general population. In this paper, we propose an analytical procedure for the SAR computation inside a biological elliptic cylinder model made up of layers of different biological tissues. The procedure makes it possible to obtain accurate SAR values in simplified models of biological subsystems, and it is also helpful to test numerical techniques to be used for more realistic models and to generate synthetic input data for diagnostic methodologies. For the assumed model, the calculation of the analytical solution has been obtained by generalizing a known procedure that deals only with lossless materials, and the model makes possible the calculation of the SAR for realistic human tissues. Various calculations prove the reliability of the technique. PMID- 10453066 TI - Influence of combined DC and AC magnetic fields on rat behavior. AB - The action of combined parallel static (DC) and alternating (AC) magnetic fields at the cyclotron frequencies for different biologically active ions, specifically, calcium, sodium, potassium, chlorine, magnesium and lithium, on rat behavior in the "open field" were investigated. It was shown that the DC and AC fields at the calcium cyclotron frequency lower the locomotor and exploratory activity of the rats, whereas action of the fields at the magnesium cyclotron frequency enhances these forms of behavioral activity. The effects were qualitatively alike at the weak (50 microT) and relatively strong (500 microT) DC fields with proportional changes in the frequencies and amplitudes of the AC fields. Statistically significant effects of cyclotron frequencies for other ions studied were not observed. PMID- 10453065 TI - Protective effect of low frequency low energy pulsing electromagnetic fields on acute experimental myocardial infarcts in rats. AB - This series of experiments assesses the effect of exposure to low-frequency pulsing electromagnetic fields (PEMFs) in 340 rats with acute experimental myocardial infarcts. The left anterior descending artery was ligated with suture thread, and the rats underwent total body exposure to PEMFs until they were killed. Twenty-four hours after surgery, the necrotic area was evaluated by staining with triphenyltetrazolium chloride. A significant reduction of the necrotic area was observed in the animals exposed to PEMFs compared with the nonexposed controls. Exposure for up to 6 days does not appear to affect the area of necrosis, although in exposed animals an increase of vascular invasion of the necrotic area is observed: 24.3 % as against 11.3 % in controls. No effect on the necrotic area size from exposure was found when the left anterior descending artery was occluded for 60 min, followed by reperfusion. The results reported show that exposure to PEMFs is able to limit the area of necrosis after an acute ischemic injury caused by permanent ligation of the left anterior descending artery. These data are in agreement with the protective effect of PEMFs observed on acute ischemia in skin free flaps in rats and in cerebral infarcts in rabbits. PMID- 10453067 TI - Effects of microscope objectives on magnetic field exposures. AB - Distortions in magnetic field intensity generated by commonly used microscope objectives (1x to 100x) were characterized within a Helmholtz coil-based exposure system. Objectives from a variety of manufacturers distorted applied field intensities by up to 23% in the image plane. Components that contribute to distortions include (1) nickel-chrome plating of objective housings, (2) the presence of steel springs in objectives with compression collars, and (3) steel screws or studs used to hold together separately manufactured parts. Steel springs and screws produce radially asymmetric profiles, whereas distortions generated by nickel-chrome plating are typically radially symmetric. All components can produce spatial gradients in field intensity if objectives are not perfectly aligned with exposure systems or if placed in the earth's magnetic field. Alterations in the magnitude of magnetic field intensities as well as the production of spatial gradients might have an effect on biological responses. By maintaining optical glass components and replacing metallic components, functional objectives can be reconstructed that produce no measurable effects on magnetic flux densities. PMID- 10453071 TI - Interaction between dexamethasone and butyrate in apoptosis induction: non additive in thymocytes and synergistic in a T cell-derived leukemia cell line. AB - In thymocytes butyrate and trichostatin A are unable to augment dexamethasone induced apoptosis. In cultured rat thymocytes the extent of apoptosis induced by dexamethasone alone did not increase by addition of 0.1 - 10 mM butyrate. Even more pronounced was the non-additive interrelationship between dexamethasone and trichostatin A, as trichostatin A-induced apoptosis was not only blocked by the presence of dexamethasone but dexamethasone-induced apoptosis was also partially inhibited in the presence of 0.1 - 0.5 microM trichostatin A. The fact that the non-additive relationship with dexamethasone for apoptosis induction was observed with both histone deacetylase inhibitors suggests that in thymocytes this phenomenon is related to histone acetylation. In contrast to this, in the human T cell-derived leukemia cell line CEM-C7H2, dexamethasone did not block butyrate- or trichostatin A-induced apoptosis; moreover, butyrate, in the concentration range of 0.1 - 1 mM, had a marked synergistic effect on dexamethasone-induced apoptosis. This synergism, however, was not mimicked by trichostatin A, indicating that the effect is not related to histone acetylation but rather due to a pleiotropic effect of butyrate. Furthermore, in CEM-C7H2 cells, at higher concentrations of butyrate (5 - 10 mM) or trichostatin A (0.4 - 0.8 microM), there was a minor but reproducible antagonistic effect of dexamethasone on apoptosis induced by each of the two histone deacetylase inhibitors, suggesting that this antagonistic effect too, is related to histone hyperacetylation. PMID- 10453070 TI - The role of Ets family transcription factor PU.1 in hematopoietic cell differentiation, proliferation and apoptosis. AB - The PU.1 gene encodes an Ets family transcription factor which controls expression of many B cell- and macrophage-specific genes. Expression of the gene is critical for development of lymphoid and myeloid cell lineages, since PU.1 deficient mice exhibit defects in the development of these cell lineages. The PU.1 gene is identical to the Spi-1 gene isolated from common proviral integration sites in Friend virus-induced murine erythroleukemia (MEL), and deregulated expression of the gene is believed to be an essential step of the disease. We recently demonstrated that overexpression of PU.1 inhibits erythroid differentiation of MEL cells induced with the differentiating agent DMSO. We also noticed unexpectedly that overexpression of PU.1 together with DMSO induces marked growth arrest and apoptosis in MEL cells, supporting the notion that some oncogenes induce growth inhibition and apoptosis rather than cell proliferation and transformation under specific circumstances as shown with the c-myc gene. In this review, the role of PU.1 in hematopoietic cell differentiation, proliferation and apoptosis is described and the possible molecular mechanisms of PU.1-induced effects in MEL cells are discussed. PMID- 10453072 TI - Ceramide accumulation precedes caspase-dependent apoptosis in CHP-100 neuroepithelioma cells exposed to the protein phosphatase inhibitor okadaic acid. AB - The protein phosphatase inhibitor okadaic acid (OA) dose-dependently induced apoptosis in CHP-100 neuroepithelioma cells when administered for 24 h at concentrations ranging from 10 - 100 nM. Apoptosis was largely, albeit not completely, dependent on cystein protease (caspase) activation. CPP32 processing and poly(ADP-ribose) polymerase (PARP) cleavage started to be observed only at 20 nM OA; moreover, the caspase inhibitor Z-Val-Ala-DL-Asp-fluoromethylketone (Z VAD.fmk) (100 microM) had negligible effect on apoptosis induced by 10 nM OA, but rescued from death an increasing cell fraction as OA concentration was raised from 20 - 100 nM. Cell treatment for 24 h with OA induced ceramide accumulation; the phenomenon started to be evident at 20 nM OA and reached its maximum at 50 - 100 nM OA. In cells exposed to 50 nM OA, ceramide was already elevated by 5 h; at this time, however, PARP cleavage and apoptosis were not yet observed. Z-VAD.fmk (100 microM) had no effect on ceramide elevation induced by 50 nM OA within 5 h, but markedly reduced ceramide accumulation as the incubation was prolonged to 24 h. The latter phenomenon was accompanied by elevation of glucosylceramide levels, thus suggesting that a caspase-dependent reduction of glucosylceramide synthesis might contribute to late ceramide accumulation. Short-chain ceramide (30 microM) induced apoptosis in CHP-100 cells and its effect was additive with that evoked by OA (10 - 20 nM). These results suggest that ceramide generation might be an important mechanism through which sustained protein phosphatase inhibition induces caspase activation and apoptosis in CHP-100 cells. PMID- 10453073 TI - Increased cell surface exposure of phosphatidylserine on propidium iodide negative thymocytes undergoing death by necrosis. AB - Phosphatidylserine (PS) exposure on propidium iodide negative cells using FITC labelled annexin-V has been used to quantify apoptosis in vitro and in vivo. Detection of PS within cells undergoing necrosis is also possible if labelled annexin-V specific for PS enters the cell following early membrane damage. Necrotic or late apoptotic cells can be excluded from flow cytometric analysis using propidium iodide which enters and stains cells with compromised membrane integrity. Here we show that thymocytes undergoing death exclusively by necrosis show early exposure of PS prior to loss of membrane integrity. This early exposure of PS occurs in cells treated with agents which both raise intracellular calcium levels and are also capable of interacting with protein thiol groups. We also demonstrate that PS exposure in thymocytes induced to undergo apoptosis by three different agents does not correlate with calcium rises but correlates with and precedes DNA fragmentation. PMID- 10453074 TI - Another cell death induction system: TNF-alpha acts as a ligand for Fas in vaginal cells. AB - The death receptor Fas transduces apoptotic death signaling upon stimulation with the Fas ligand. We previously reported that Fas contributes to vaginal cell death observed during the estrus cycle and after estrogen deprivation, using the functional Fas-lacking lpr and lprcg mutant mouse. In the present study, we investigated whether the Fas ligand also plays a dominant role in vaginal cell death using the functional Fas ligand-lacking gld mutant mouse. Our results demonstrated that vaginal cells of gld mice do not show any abnormalities, suggesting the possible presence of another ligand for Fas. Through our investigation, we demonstrated TNF-alpha as a ligand for vaginal Fas. Here, we propose that TNF-alpha acts for the ligand for Fas in vaginal cells, suggesting a new cell death induction system. PMID- 10453075 TI - Caspases: their intracellular localization and translocation during apoptosis. AB - The activation of the caspase family of proteases has been detected in numerous cell systems and appears to function as a common pathway through which apoptotic mechanisms may operate. Caspases are synthesized as precursors (pro-caspases) and are converted into mature enzymes by apoptotic signals. The effects of caspases in apoptosis are accomplished by the cleavage of numerous proteins located in different intracellular compartments. In the present study we have addressed the question of the subcellular localization of different pro- and active caspases as well as several other proteins, such as Apaf-1, calpain and DFF, which also play important roles in the apoptotic process. We found that at least three pro caspases (pro-caspases-2, -3 and -9) were present in both the mitochondrial and cytosolic fractions of untreated Jurkat T lymphocytes. Only pro-caspase-2 was found in the nuclear fraction. Pro-caspases-7 and -8 were found only in the cytosolic fraction. In apoptotic cells, caspases-3, -8 and -9 were present in the cytosolic fraction, whereas caspases-3 and -9 were also found in the mitochondrial fraction and caspase-7 in the microsomal fraction. Caspases-2 and 3 were present in the nuclear fraction. The selective localization of pro caspases in different subcellular compartments may play an important, but yet unknown, role in their activation. The translocation of active caspases to other subcellular compartments appears to be critical for the development of the apoptotic process. PMID- 10453076 TI - Induction of apoptosis by IFNgamma in human neuroblastoma cell lines through the CD95/CD95L autocrine circuit. AB - The CD95 (APO-1/Fas) system can mediate apoptosis in immune cells as well as in tumour cells, where it may contribute to tumour immune-escape. On the other hand, its induction by anticancer drugs may lead to tumour reduction. Interferongamma (IFNgamma) increases the sensitivity of tumour cell lines to anti-CD95 antibody mediated apoptosis. We describe induction of apoptosis by IFNgamma through the expression of CD95 and its ligand (CD95L) in human neuroblastoma cell lines. Neuroblastoma cells showed low constitutive expression of CD95 and CD95L. Subsequent to IFNgamma-modulated increase in CD95 and CD95L mRNA as well as protein levels, apoptosis was observed. Our results demonstrated that cytokine mediated apoptosis was mediated through the activation of the CD95/CD95L autocrine circuit since: (i) cell death occurred following CD95/CD95L expression and correlated with CD95 and CD95L expression levels, (ii) failed to occur in a clone which weakly upregulated CD95 and lacked CD95L induction after IFNgamma stimulation, (iii) was at least partially inhibited by using blocking F(ab')2 anti-CD95 antibody fragments and the recombinant Fas-Fc protein, that prevented the interaction between CD95 and CD95L. The intracellular molecular mechanisms elicited by IFNgamma are clearly highly complex, with several signalling pathways being activated, including the CD95 system. These findings suggest that IFNgamma may have a significant potential in the therapy of neuroblastoma in vivo. PMID- 10453077 TI - Activation of cAMP signaling transiently inhibits apoptosis in vascular smooth muscle cells in a site upstream of caspase-3. AB - Intracellular signaling pathways that are involved in protection of vascular smooth muscle cells (VSMC) from apoptosis remain poorly understood. This study examines the effect of activators of cAMP/cGMP signaling on apoptosis in non transfected VSMC and in VSMC transfected with c-myc (VSMC-MYC) or with its functional analogue, E1A-adenoviral protein (VSMC-E1A). Serum-deprived VSMC-E1A exhibited the highest apoptosis measured as the content of chromatin and low molecular weight DNA fragments, phosphatidylserine content in the outer surface of plasma membrane and caspase-3 activity (ten-, five-, four- and tenfold increase after 6 h of serum withdrawal, respectively). In VSMC-E1A, the addition of an activator of adenylate cyclase, forskolin, abolished chromatin cleavage, DNA laddering, caspase-3 activation and the appearance of morphologically-defined apoptotic cells triggered by 6 h of serum deprivation. In non-transfected VSMC and in VSMC-MYC, 6 h serum deprivation led to approximately six- and threefold activation of chromatin cleavage, respectively, that was also blocked by forskolin. In VSMC-E1A, inhibition of apoptosis was observed with other activators of cAMP signaling (cholera toxin, isoproterenol, adenosine, 8-Br cAMP), whereas 6 h incubation with modulators of cGMP signaling (8-Br-cGMP, nitroprusside, atrial natriuretic peptide, L-NAME) did not affect the development of apoptotic machinery. The antiapoptotic effect of forskolin was abolished in 24 h of serum deprivation that was accompanied by normalization of intracellular cAMP content and protein kinase A (PKA) activity. Protection of VSMC-E1A from apoptosis by forskolin was blunted by PKA inhibitors (H-89 and KT5720), whereas transfection of cells with PKA catalytic subunit attenuated apoptosis triggered by serum withdrawal. The protection of VSMC-E1A by forskolin from apoptosis was insensitive to modulators of cytoskeleton assembly (cytochalasin B, colchicine). Neither acute (30 min) nor chronic (24 h) exposure of VSMC to forskolin modified basal and serum-induced phosphorylation of the MAP kinase ERK1/2. Thus, our results show that activation of cAMP signaling delays the development of apoptosis in serum-deprived VSMC at a site upstream of caspase-3 via activation of PKA and independently of cAMP-induced reorganization of the cytoskeleton network and the ERK1/2-terminated MAPK signaling cascade. PMID- 10453078 TI - Bax antisense oligonucleotides reduce axotomy-induced retinal ganglion cell death in vivo by reduction of Bax protein expression. AB - Following transection of the optic nerve (ON), retinal ganglion cells (RGCs) upregulate Bax protein expression and undergo apoptosis. The present study aimed at reducing Bax expression in order to test whether Bax plays a causative role in the induction of secondary RGC apoptosis. Following injection into the vitreous, fluoresceinated oligonucleotides transfected RGCs in vivo at the injection site in the temporal superior retina. Following ON lesion, and repeated injections of a partially phosphorothioated Bax antisense oligonucleotide, but not following injection of control oligonucleotides, expression of Bax protein was locally inhibited, and the number of surviving RGCs was increased in Bax antisense treated rats 8 days after axotomy. Our results indicate that Bax induction is a prerequisite for the execution of RGC apoptosis following ON axotomy. While the Bax antisense strategy offers an exciting perspective to inhibit secondary neuronal degeneration in vivo, both limited transfection efficacy, and the temporal restriction of this effect currently limit the use of this approach with respect to clinical applications for the treatment of neurodegeneration. PMID- 10453079 TI - Analysis of redox regulation of cytochrome c-induced apoptosis in a cell-free system. AB - In this study, we investigated the importance of redox and Bcl-2 status on cytochrome c-mediated apoptosis. Two mouse lymphoma cell lines, LYas and LYar that express Bcl-2 protein at different levels, were used to reconstitute a cell free system. Cytoplasmic extracts made from apoptosis-sensitive LYas cells 2.5 h after exposure to 5 Gy gamma-radiation were able to induce apoptosis in isolated nuclei, whereas extracts made from LYas cells at time points earlier than 2. 5 h, or from Bcl-2-overexpressing, apoptosis-resistant LYar cells at all time points after irradiation were inactive. Apoptotic activity was restored to inactive extracts by the addition of oxidized but not reduced cytochrome c. Cytochrome c reductase was able to inhibit apoptosis in extracts made from LYas cells 2.5 h after irradiation and LYar extracts activated by addition of oxidized cytochrome c. Antioxidants, but not oxidant defensive enzymes, blocked apoptosis implying that antioxidants might alter the redox state of factors important in mediating apoptosis. These findings confirm the importance of cellular redox state during apoptosis and are consistent with a role for Bcl-2 in regulating this redox state. PMID- 10453080 TI - Nuclear localization of a new c-cbl related protein, CARP 90, during in vivo thymic apoptosis in mice. AB - This study investigates the involvement of the c-cbl protooncogene in thymocyte apoptosis occurring in vivo after hydrocortisone treatment. In the thymus of untreated mice, a few medullary and cortical thymocytes expressed p120cbl, mainly in the cytoplasm. In the cortex, their number and distribution resemble that of apoptotic cells evidenced by TUNEL staining. The expression of Cbl is rapidly increased when apoptosis is triggered by hydrocortisone. This Cbl-specific immunostaining was detected in the nucleus and is due to a Cbl-related 90 kDa protein (CARP 90). These results show that a c-cbl product could localize in the nucleus and suggest that it could be involved as a regulator of thymic apoptosis. PMID- 10453081 TI - Modulation of apoptosis of proliferating and differentiating HL-60 cells by protein kinase inhibitors: suppression of PKC or PKA differently affects cell differentiation and apoptosis. AB - The relationship between RA- or dbcaMP-mediated differentiation and subsequent apoptosis in HL-60 cells was assessed by modulating the levels of differentiation suppressing the activity of PKC and PKA with calphostin C or GF 109203X and H89, respectively. Results demonstrated that (1) RA and dbcAMP caused a dose-dependent increase in apoptosis concomitant with progressive differentiation; (2) the suppression of PKC activity resulted in an increase of apoptosis unrelated to the modulated levels of differentiation; (3) the inhibition of PKA decreased granulocytic differentiation, but did not significantly affect apoptosis; (4) the pretreatment of cells with dbcAMP strongly potentiated RA-mediated differentiation without apparent changes in apoptosis; (5) cell differentiation and apoptosis were associated with cell cycle arrest in G1 phase and G2/M phases, respectively. Our findings indicate that the functional maturity of differentiating cells is not directly related to the apoptotic programme, and suggest that induction of cell differentiation and apoptosis are regulated by separate mechanisms in which PKC and PKA are involved. PMID- 10453082 TI - Induction of stress genes by low doses of gamma rays. AB - Using cells of a human myeloid tumor cell line (ML-1), we have detected induction of several stress-responsive genes by doses of gamma rays below 50 cGy. We found a linear dose-response relationship for induction of CDKN1A (formerly known as CIP1/WAF1) and GADD45 mRNA levels over the range of 2-50 cGy, with no evidence of a threshold for induction. Although exposures to 2 and 5 cGy did not result in any detectable reduction in cloning efficiency or increased apoptosis in ML-1 cells, these exposures did produce a transient delay of cells in the phases of the cell cycle in addition to the observed up-regulation of CDKN1A and GADD45. The relative induction of genes such as CDKN1A by radiation doses that produce little toxicity indicates that surviving cells do contribute significantly to the observed stress responses. These studies should provide insight into the molecular responses to physiologically relevant doses that cannot necessarily be extrapolated from high-dose studies. PMID- 10453083 TI - A dose response for radiation-induced intrachromosomal DNA rearrangements detected by inverse polymerase chain reaction. AB - X-ray-induced intrachromosomal DNA rearrangements were detected in the 5' region of the MYC gene of cells of the human bladder carcinoma cell line, EJ-30, by using PCR with inverted primers. When the cells were allowed to repair/misrepair for 6 or 23 h after irradiation, the frequency of rearrangements increased with dose from (0.7 +/- 0.4) x 10(-5) per copy of MYC for unirradiated cells to (3.2 +/- 0.7) x 10(-5) after 30 Gy, (5.4 +/- 1.2) x 10(-5) after 70 Gy, and (5.9 +/- 1.0) x 10(-5) after 100 Gy. No significant difference was observed between 6 and 23 h of repair. Sequences obtained from the products suggest that there was no homology between the two sequences involved in the recombination event and that there was no clustering of breakpoints. The procedure is relatively simple, requiring only one digestion with a rare-cutting restriction enzyme prior to PCR amplification of the DNA purified from irradiated cells. The site of enzyme digestion is located between a pair of primer sites 120 bp apart for which the primers face in opposite directions. If no intrachromosomal rearrangement has occurred, no PCR product would be obtained. However, if an intrachromosomal rearrangement has occurred between two regions located on either side of the primer sites, an episome or duplication event would result if the rearrangement had occurred either within the same chromatid or between two sister chromatids, respectively. Digestion between the primers would linearize an episome or release a linear molecule containing the duplicated primer sites from a larger molecule. After both types of rearrangement events, the primers would be facing each other and would be located on either end of the linear molecule; and if they are less than approximately 5 kb apart, PCR amplification should result in a product. This procedure is relatively simple and rapid and does not require any cell division after irradiation or phenotypic selection of mutants. Also, quantification is based on the number of PCR products detected in a known amount of DNA, and not on a precise determination of the amount of PCR amplification that has occurred. Thus the inverse PCR procedure has the potential ofbeing used as an assay to detect variations in radiation-induced frequencies of DNA rearrangements. PMID- 10453084 TI - Regional tumor oxygenation and measurement of dynamic changes. AB - We recently described a novel approach to measuring regional tumor oxygen tension using (19)F pulse burst saturation recovery (PBSR) nuclear magnetic resonance (NMR) echo planar imaging (EPI) relaxometry of hexafluorobenzene. We now compare oxygen tension measurements in a group of size-matched R3327-AT1 Dunning prostate rat tumors made using this new method with those using a traditional polarographic method: the Eppendorf histograph. Similar oxygen tension distributions were found using the two methods, and both techniques showed that tumors with volume greater than 3.5 cm(3) were significantly (P < 0.0001) less well oxygenated than smaller tumors (volume less than 2 cm(3)). Using the (19)F EPI approach, we also examined the response to respiratory challenge. Increasing the concentration of inspired oxygen from 33% to 100% O(2) produced a significant increase (P < 0.0001) in tumor oxygenation for a group of small tumors. In contrast, no change was observed in the mean pO(2) for a group of large tumors. Consideration of individual tumor regions irrespective of tumor size showed a strong correlation between the maximum pO(2) observed when breathing 100% O(2) compared with mean baseline pO(2). These results further demonstrate the usefulness of (19)F EPI to assess changes in regional tumor oxygenation. PMID- 10453085 TI - Incorporation of an (125)I-labeled hexa-iodinated diglyceride analog into low density lipoprotein and high specific uptake by cells of cervical carcinoma cell lines. AB - The feasibility of using low-density lipoprotein (LDL) to deliver cytotoxic drugs to tumor cells has been explored since the 1980s, when cells of a number of cancer cell lines were found to have higher LDL receptor activity than normal cells. Such differential uptake between tumor and normal cells may provide a unique opportunity to use LDL as a tumor-specific carrier of radiopharmaceuticals for the clinical management of cancer. In this study, an (125)I-labeled hexa iodinated diglyceride analog, 1, 3-dihydroxypropan-2-one 1,3-diiopanoate (DPIP), was synthesized and incorporated into LDL using a fusion technique. It was found that approximately 500 [(125)I] DPIP molecules were incorporated into each LDL particle. Cells of three human cervical tumor cell lines, HeLa, SiHa and C-33A, were used to examine the cellular uptake of the [(125)I]DPIP-LDL conjugate. It was shown that the [(125)I]DPIP-LDL conjugate was specifically bound to and taken up by cervical tumor cells through an LDL receptor-mediated endocytosis pathway. The results suggest that LDL may be a selective carrier for delivering hydrophobic radiopharmaceuticals to cancer cells and particularly for the diagnosis of cervical tumors. PMID- 10453086 TI - Effect of radiation on the xanthine oxidoreductase system in the liver of mice. AB - The xanthine oxidoreductase system is one of the major sources of free radicals in many pathophysiological conditions. Since ionizing radiations cause cell damage and death, the xanthine oxidoreductase system may contribute to the detrimental effects in irradiated systems. Therefore, modulation of the xanthine oxidoreductase system by radiation has been examined in the present study. Female Swiss albino mice (7-8 weeks old) were irradiated with gamma rays (1-9 Gy) at a dose rate of 0.023 Gy s(-1) and the specific activities of xanthine oxidase (XO) and xanthine dehydrogenase (XDH) were determined in the liver of the animals. The mode and magnitude of change in the specific activities of XO and XDH were found to depend on radiation dose. At doses above 3 Gy, the specific activity of XO increased rapidly and continued to increase with increasing dose. However, the specific activity of XDH was decreased. These findings are suggestive of an inverse relationship between the activity of XO and XDH. The ratio of the activity of XDH to that of XO decreased with radiation dose. However, the total activity (XDH + XO) remained constant at all doses. These results indicate that XDH may be converted into XO. An intermediate form, D/O, appears to be transient in the process of conversion. The enhanced specific activity of XO may cause oxidative stress that contributes to the radiation damage and its persistence in the postirradiation period. Radiation-induced peroxidative damage determined in terms of the formation of TBARS and the change in the specific activity of lactate dehydrogenase support this possibility. PMID- 10453087 TI - Effects of prenatal low-dose beta radiation from tritiated water on learning and memory in rats and their possible mechanisms. AB - Pregnant adult Wistar rats were randomly divided into four groups. Three of these groups were irradiated with beta rays by a single intraperitoneal injection of tritiated water ((3)H(2)O) administered on the 13th day of gestation. The doses absorbed by their offspring were estimated to be 4.6, 9.2 and 27.3 cGy. The influence of radiation on the postnatal learning ability and memory behavior and on brain development of the offspring was investigated. The number of pyramidal cells (in areas CA1, CA2, CA3 and CA4) and neurons in the hippocampus of the offspring was also measured. In addition, the Ca(++) conductance of hippocampal pyramidal cells cultured in vitro was observed. The results showed that an exposure to 4.6 cGy could prolong avoidance response time significantly and decrease the number of hippocampal pyramidal cells in the CA1 area compared to controls. An exposure to 9.2 cGy significantly decreased the establishment of conditioned reflexes and the number of hippocampal pyramidal cells in the CA3 area. This exposure also induced the degeneration and malformation of hippocampal neurons cultured in vitro, in addition to decreasing the number of hippocampal neurons observed on each culture day. A dose of 27.3 cGy significantly decreased brain and body weights and the maximum electric conductance of Ca(++) in hippocampal pyramidal neurons. In general, dose-dependent effects were observed for most of the parameters assessed in the present study. Possible mechanisms are discussed. PMID- 10453088 TI - The adaptive response modifies latency for radiation-induced myeloid leukemia in CBA/H mice. AB - We have investigated the effect of the adaptive response on acute myeloid leukemia (AML) induced in CBA/Harwell mice by a chronic radiation exposure. Groups of mice irradiated with a total dose of 1. 0 Gy at two different chronic dose rates (0.5, 0.004 Gy/h) had similar frequencies of AML. Compared to control animals that did not develop AML, irradiation at either of these dose rates did not change the longevity of the mice that did not die of leukemia. The survival rates of irradiated mice that did develop leukemia in the two groups were not different from each other, indicating that the dose rates produced similar responses and therefore were both chronic exposures. We then tested the ability of a chronic 10-cGy (0. 5 Gy/h) exposure to ionizing radiation, mild hyperthermia (40.5 degrees C whole-body, 60 min) or treatment with interleukin-1 (1500 U i.p.) to induce an adaptive response and modify the frequency or latency of AML which resulted from a subsequent (24 h later) 1.0-Gy (0.5 Gy/h) chronic radiation exposure. The frequency of radiation-induced leukemia was not changed in mice given any of the three adapting treatments 24 h prior to the chronic 1.0-Gy dose that induced leukemia. However, the latent period for development of AML was significantly increased by both the prior low radiation dose and mild hyperthermia treatment. Injection of interleukin-1, in contrast, may have reduced the latent period. Similar to the single 1.0-Gy chronic exposure alone, none of the adapting treatments prior to that exposure influenced the survival of animals that did not develop AML. These results indicate that an earlier exposure to a small adapting dose of radiation or to a mild heat stress can influence secondary steps in radiation-induced carcinogenesis. PMID- 10453089 TI - Risks of leukemia in Japanese atomic bomb survivors, in women treated for cervical cancer, and in patients treated for ankylosing spondylitis. AB - The dose-response relationship for radiation-induced leukemia was examined in a pooled analysis of three exposed populations: Japanese atomic bomb survivors, women treated for cervical cancer, and patients irradiated for ankylosing spondylitis. A total of 383 leukemias were observed among 283,139 study subjects. Considering all leukemias apart from chronic lymphocytic leukemia, the optimal relative risk model had a dose response with a purely quadratic term representing induction and an exponential term consistent with cell sterilization at high doses; the addition of a linear induction term did not improve the fit of the model. The relative risk decreased with increasing time since exposure and increasing attained age, and there were significant (P < 0.00001) differences in the parameters of the model between datasets. These differences were related in part to the significant differences (P = 0.003) between the models fitted to the three main radiogenic leukemia subtypes (acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia). When the three datasets were considered together but the analysis was repeated separately for the three leukemia subtypes, for each subtype the optimal model included quadratic and exponential terms in dose. For acute myeloid leukemia and chronic myeloid leukemia, there were reductions of relative risk with increasing time after exposure, whereas for acute lymphocytic leukemia the relative risk decreased with increasing attained age. For each leukemia subtype considered separately, there was no indication of a difference between the studies in the relative risk and its distribution as a function of dose, age and time (P > 0.10 for all three subtypes). The nonsignificant indications of differences between the three datasets when leukemia subtypes were considered separately may be explained by random variation, although a contribution from differences in exposure dose-rate regimens, inhomogeneous dose distribution within the bone marrow, inadequate adjustment forcell sterilization effects, or errors in dosimetry could have played a role. PMID- 10453090 TI - Spontaneous and nitrosourea-induced primary tumors of the central nervous system in Fischer 344 rats chronically exposed to 836 MHz modulated microwaves. AB - We have tested an 836.55 MHz field with North American Digital Cellular (NADC) modulation in a 2-year animal bioassay that included fetal exposure. In offspring of pregnant Fischer 344 rats, we tested both spontaneous tumorigenicity and the incidence of induced central nervous system (CNS) tumors after a single dose of the carcinogen ethylnitrosourea (ENU) in utero, followed by intermittent digital phone field exposure for 24 months. Far-field exposures began on gestational day 19 and continued until weaning at age 21 days. Near-field exposures began at 35 days and continued for the next 22 months, 4 consecutive days weekly, 2 h/day. SAR levels simulated localized peak brain exposures of a cell phone user. Of the 236 original rats, 182 (77%) survived to the termination of the whole experiment and were sacrificed at age 709-712 days. The 54 rats (23%) that died during the study ("preterm rats") formed a separate group for some statistical analyses. There was no evidence of tumorigenic effects in the CNS from exposure to the TDMA field. However, some evidence of tumor-inhibiting effects of TDMA exposure was apparent. Overall, the TDMA field-exposed animals exhibited trends toward a reduced incidence of spontaneous CNS tumors (P < 0. 16, two-tailed) and ENU induced CNS tumors (P < 0.16, two-tailed). In preterm rats, where primary neural tumors were determined to be the cause of death, fields decreased the incidence of ENU-induced tumors (P < 0.03, two-tailed). We discuss a possible approach to evaluating with greater certainty the possible inhibitory effects of TDMA-field exposure on tumorigenesis in the CNS. PMID- 10453091 TI - The UVA light used during the fluorescence microscopy assay affects the level of intracellular calcium being measured in experiments with electric-field exposure. AB - In the present paper, the induction of calcium signals in neuroblastoma cells, cells of T-cell leukemia, and osteogenic sarcoma cells were investigated in relation to the UVA irradiation used in fluorescence microscopy. Methods were developed to measure both the mean UVA irradiance and the intensity profile in the UVA-illuminated area of the microscope. This allowed us to calculate the applied UVA radiant exposure of the cells during each experiment. This investigation was undertaken because of the conflicting results in the literature on the effects of electromagnetic fields on the signals of the calcium-sensitive fluorescence probe FURA-2 in lymphocytes. Taking into account that each group used a different system with different optics and lamps, these conflicting results are now at least partially understandable. Our measurements indicate that in a typical experiment with FURA-2 the cells were irradiated with up to 776 kJ m(-2) during 25 min of exposure to UVA light. This causes changes in intracellular free Ca(2+) concentrations ([Ca(2+)](i)). Designating cells in which the [Ca(2+)](i) was distinctly increased during the experiment as "responding", we found Hill-type dependences on the irradiance. Jurkat cells showed a 50% response even at 10 kJ m(-2) and osteosarcoma cells at about 60 kJ m(-2), whereas neuroblastoma cells even at the maximum possible dose responded only minimally. In the case of neuroblastoma cells, we found a dependence of this effect on the CO(2) partial pressure during the preincubation. An electrical treatment with an a.c. field (5 kHz sinusoidal, amplitude modulation 16 Hz 100%, 800 V m(-1), 5 min) had a significant effect on intracellular calcium in neuroblastoma cells only in the case of cells that were not pretreated with CO(2) with high fluences of UVA irradiation. In conclusion, these results indicate that the possibility of UVA artifacts must be considered in all experiments using fluorescence microscopy. Furthermore, our results lead to the hypothesis that oxidative stress could be the link between UVA and electric-field effects. PMID- 10453092 TI - On the assumption of negligible heat diffusion during the thermal measurement of a nonuniform specific absorption rate. AB - The problem of measuring a spatially varying specific absorption rate (SAR) by thermal techniques is treated both analytically and numerically. It is shown that, unless the measurement is attempted at an inflection point of the SAR distribution, it will be confounded by thermal diffusion within the medium. Rules of thumb are provided to enable the experimenter to gauge the thermal conduction contributions (i.e. error) to the thermally determined SAR near a spatial extremum. The simplest of these is that the width t(m) [s] of the time window, over which temperature variation associated with SAR is measured, should satisfy the inequality t(m) or = 140/90 mm Hg) for participants aged 25 years and older was 7-8% in Idere and 24 27% in Idikan. The distributions of overweight, sodium/potassium ratio, perceived stress, and social integration scores all contributed to lower hypertension risk in Idere. The effects and interactions of these identified risk factors remain poorly understood, even among people who share a common genetic background, similar diet, and many other lifestyle features. Nonetheless, the rural-urban distinction is sufficiently salient to engender a nearly threefold difference in hypertension prevalence. This disparity in disease prevalence demonstrates the sensitivity of human beings to the environmental determinants of disease and provides a sobering example of the difficulty in identifying subtle genetic effects, which can be easily overwhelmed by small differences in environmental exposures. PMID- 10453106 TI - Blood pressure in the Caribbean. AB - A review of original and published data yields the consistent finding that chronic hypertension is endemic and prevalent throughout the Caribbean area. The prominent involvement of genetics is suggested in several lines of evidence, including significant ethnic differences in blood pressure values, higher incidence in smaller isolated island populations, the discovery of polymorphic blood proteins that appear to be linked to hypertension susceptibility, and the slavery hypothesis of natural selection favoring a salt-conserving physiology in ancestral populations. Environmental factors--climatic, demographic, and cultural -exert strong influences on blood pressure levels and hypertension etiology in the Caribbean. Salt intake and other dietary behaviors, degree of community awareness of the disease, and differential treatment modalities are related to hypertension epidemiology in indigenous and migrant Caribbean populations. The traditional use of medicinal plants, historically successful in part because of the beneficial bioactivity of many antihypertensive phytochemical components, has been recently supplemented with the widespread introduction of synthetic biomedical drugs. Prospective research strategies are recommended that might further elucidate the complex gene-environment interactions contributing to blood pressure variation and hypertension patterns in the Caribbean region. PMID- 10453107 TI - Heterogeneity of genetic control of blood pressure in ethnically different populations. AB - We review the literature on statistical genetic analyses of blood pressure in samples from various ethnic backgrounds using different statistical methods and packages. We then provide the results of a complex segregation analysis performed on familial data on systolic and diastolic blood pressure in 2 ethnically different populations, Chuvashans and Turkmenians. Two types of major gene models were tested in the segregation analysis: Model type 1 tests for a Mendelian mode of transmission and estimates genotype-specific averages regardless of age and sex effect, and model type 2 estimates age and sex effects on each of 3 genotypes within the putative major genotype. In both total samples, by both types of segregation analysis, familial aggregation of both systolic and diastolic blood pressure was inconsistent with the Mendelian mode of inheritance. In the next step of analysis the pedigrees in both samples were sorted into 2 groups on the basis of 2 likelihoods as obtained under Mendelian and nontransmission models for each entire sample. This procedure resulted in the appearance of 2 subsamples (large and small) in each ethnic sample. The segregation analysis that was carried out then on the larger subsample provided consistent evidence to support the major gene effect on systolic and diastolic blood pressure in 2 ethnic groups. Interestingly, model type 2 showed that in both ethnically different large subsamples, for each sex the genotype predisposing to a larger mean value of systolic (or diastolic) blood pressure also displayed the highest rate of blood pressure increase with age. We discuss in detail possible sources of heterogeneity in familial transmission of blood pressure observed in our 2 samples, and we suggest a method to improve the analysis of heterogeneity for trait inheritance. PMID- 10453108 TI - The long good Friday: NCHD hours. PMID- 10453109 TI - Medical care of athletes in the era of performance enhancing drugs of sport (PEDOS) syndrome. AB - We have arrived at a watershed in the fight against drugs in sport. This must not be looked upon as a war. War eventually has winners and losers. In the fight against drugs in sport we have a major ethical, educational, financial, health and management problem. The solution to the problem will be complex. At this moment in time there is a suggestion that we move away from sports administering the problem, to governments doing the job of controlling drugs in sport. We must remind ourselves that there is no track record for governments doing the job. Perhaps representatives from the two groups, to include athletes, sports administrators, politicians, doctors with an interest in pharmacology, endocrinology, respiratory medicine, sport, and rehabilitation, pharmacists, lawyers, medical and political administrators and laboratory personnel, under the chairmanship of a compassionate person working together may be the answer. PMID- 10453111 TI - Clinical assessment and management of hayfever in general practice. PMID- 10453110 TI - Medicine, the media and doctors under the spotlight. PMID- 10453112 TI - Manpower in respiratory medicine: a position paper of the Irish Thoracic Society. PMID- 10453113 TI - Irish health care Web sites: a review. AB - The quality of health information on the World Wide Web is ver variable. There are no accepted standards for publishing health information on the Internet. Core standards for health Web sites include: authorship, attribution, disclosure and currency. Irish health care Web sites are disappointing. They are poorly designed. In the main, no target audience is specified, no details are given of dates that information is posted or updated and no e-mail contact addresses are available. There is a paucity of health information and lack of details on authorship and attribution of the information that is available. Information on services, aimed at a public audience, has an excessively high reading age. Sites provide static information and are not interactive. Little maintenance and updating of sites takes place and many are out of date. PMID- 10453114 TI - The Irish National Centre of Pharmacoeconomics: its rationale and role. PMID- 10453116 TI - Civil justice reforms in England: the Woolf Report and the Pre-Action Protocol. PMID- 10453115 TI - Cerebral vasculitis and haemorrhage in a HIV positive intravenous drug abuser. PMID- 10453117 TI - Aggressive angiomyxoma. PMID- 10453118 TI - Myocardial infarction in a 14 year old boy after butane inhalation. PMID- 10453119 TI - Visual hallucinations associated with Charles Bonnet Syndrome--an ever increasing diagnosis. PMID- 10453120 TI - Knowledge and practices of preventing diarrhoea in malnourished children. AB - Instructions for preventing diarrhoea, based on a knowledge-deficit model, are a common health-promotion approach aimed at the providers of child care attending nutritional rehabilitation centres. However, there is rarely an assessment of baseline knowledge to justify the need for this type of intervention and to guide its form. This study investigated the practice and knowledge of recommended diarrhoea-prevention behaviours of caregivers of 78 malnourished children consecutively admitted to a realimentation programme. Major deficits included: 39% not boiling (or not planning on boiling) drinking water after the child reached two years of age; 35% not always washing children's hands before meals; only 17% reporting that it was rare for their children to go barefoot; and the majority breastfeeding for less than one year. However, almost all measures of knowledge, based on open and closed questions, were not related to the corresponding practice. Several types of barriers to preventive practices were reported on open questions, including, "beliefs," "children as barriers," and "time." This information may be helpful in designing more effective health promotion programmes. PMID- 10453121 TI - Environmental and personal hygiene practices: risk factors for diarrhoea among children of Nigerian market women. AB - A Cross-sectional survey was carried out to determine the environmental and personal hygiene practices of mothers of children aged less than five years in two markets in Ibadan--one with poor sanitary conditions (Bodija) and the other one with better sanitation facilities (Gbagi). The study sought to identify the risk factors for diarrhoea among these children. Two hundred and sixty-six mothers in Bodija and 260 in Gbagi were interviewed. A questionnaire was used for collecting information on social and demographic characteristics, personal and environmental hygiene practices, including sources of food and water for their children, waste-disposal practices and occurrence of diarrhoea among their children aged less than five years. The educational status of the women in Bodija was lower than that of the women in Gbagi (p < 0.001). Sixty (23%) women of the Bodija market mentioned that tap water was the source of drinking water for their children, while 91 (34%) brought water from their homes, and 45 (17%) bought it from vendors in the market. The corresponding figures for women of the Gbagi market were 41 (16%), 98 (38%) and 19 (7%). Two hundred and thirty-four (90%) women in Gbagi prepared breakfast at home for their children compared to 216 (81%) women in Bodija. This difference was statistically significant (p < 0.05). Waste disposal and personal hygiene practices were poorer among the women in Bodija. Yet the occurrence of diarrhoea was not significantly different in both the markets. Risk factors for diarrhoea identified in this study were water and food bought from vendors, child defaecation practices, mothers' cleaning up practices after child's defaecation, and refuse-disposal practices. The inherent risk of sale of unwholesome food and water by vendors is a great concern for public health authorities in Nigeria. Efforts to control diarrhoea must not only be focused on improving mothers' knowledge about food hygiene but also on environmental hygiene practices within the community. PMID- 10453122 TI - Detection of Shigellae from stools of dysentery patients by culture and polymerase chain reaction techniques. AB - In Bangladesh, the isolation rates of Shigella spp. range from 11% to 12% by the conventional culture technique. Since the sensitivity of this technique is low, the polymerase chain reaction (PCR) technique was used for detecting small number of Shigellae from patients' stools. Sensitivity and specificity of the two techniques were also compared. Stool samples were collected from 41 patients with dysentery who attended the Clinical Research and Service Centre of the ICDDR,B: Centre for Health and Population Research. All stool specimens were directly plated onto MacConkey, Salmonella-Shigella, Xylose lysin deoxycholate and Hectoen enteric agar media, and Shigellae were detected following standard procedures. DNA was extracted from the stool samples, and the target sequence of invasive plasmid antigen (ipa)H locus was amplified by PCR with 130 ng each of two primers (primer H8 [5'-GTTCCTTGACCGCCTTTCCGATAC-3'] and primer H15 [5'-GCCGGTCAGCCACCCTA 3']) following standard procedures. The amplified product was hybridized using an ipaH probe. The isolation rates of Shigella dysenteriae type 1, S. flexneri, S. sonnei, and S. boydii were, respectively, 17.1%, 19.5%, 4.9% and 2.4% by the conventional method. The results of the PCR technique showed that 700 bp fragment was generated in 18 of the 18 culture-positive and in 7 of the 23 culture negative stools. One hundred twenty-three strains of Escherichia coli were also tested by PCR for identifying the enteroinvasive E. coli, but none of them yielded any positive result. This study showed that the sensitivity of the culture technique is 72% and specificity is 100%, when the PCR technique was considered as gold standard. Therefore, the PCR may be considered a more sensitive and specific technique than the conventional culture technique and has the potential to be employed in routine diagnosis of dysentery in clinical centres as well as in epidemiologic studies. PMID- 10453123 TI - Use of soursop and sweetsop juice in the management of diarrhoea in children. PMID- 10453124 TI - Cyclospora cayetanensis-associated diarrhoea in a patient with acute myeloid leukaemia. PMID- 10453125 TI - Evidence of involvement of guanylate cyclase in the prolongation of caeco-colonic transit time by indomethacin in mice. PMID- 10453126 TI - Predicting tumor metastasis in patients with oral cancer by means of the proliferation marker Ki67. AB - Recent developments of a Ki67 antibody to recombinant parts of the Ki67 nuclear antigen have provided a marker for tumor proliferation. In the present study, biopsy specimens were obtained from 20 patents with squamous cell carcinoma of the oral cavity at various sites, who also received a regional neck dissection. The patients' mean age was 61 years. Normal mucosa obtained from the surgical materials of 10 patients with a non-tumor condition was also examined as a control. The expression of Ki67 was examined immunohistochemically and the labeling index (LI) assessed in the biopsy specimens. The patients were divided into two groups; patient who already had a regional lymph node metastasis at the time of tumor resection and patient without any metastasis. All of the oral carcinoma and normal mucosa specimens were positive for Ki67, while the magnitude of staining showed a wide variation. The median LI of the patients with metastasis and without metastasis was 37.63 +/- 8.30 and 20.40 +/- 4.22 respectively, while the normal mucosa control was 7.62 +/- 1.70. The results of this study suggest that an immunohistochemical examination of the biopsy materials for the Ki67 antigen and assessed LI index should prove useful for the prediction of lymph node metastasis. PMID- 10453127 TI - Evaluation of initial attachment of human gingival fibroblast cells to biodegradable membranes in vitro by light and scanning electron microscopy. AB - Guided tissue regeneration procedures using resorbable membranes have become accepted therapy for treating periodontal defects. Resorbable collagen and synthetic polylactide and polyglycolide copolymer membranes have been found to support regeneration and preclude the need for surgical removal. This study was undertaken to assess and compare the initial attachment of human gingival fibroblast cells to four collagen-based membranes (fascia lata, fascia temporalis, dura mater, and Type I bovine collagen) and a synthetic polylactic acid-based membrane (resolut). Human gingival fibroblasts were grown from explants of normal tissue obtained during surgical reduction of retromolar tissues. Membrane specimens were placed in separate culture wells and incubated with fibroblasts for one hour. The number of adherent cells was evaluated by light microscopy using an ocular grid system and detailed examination was performed by scanning electron microscopy. The results of evaluation by light microscopy indicated that initial cell attachment was significantly less in the polylactic acid-based membrane group than in the collagen-based membrane groups (P < 0.01). However, no significant differences were found among the collagen membrane groups in terms of fibroblast attachment (P > 0.01). Scanning electron microscopy examination of fibroblasts cultured directly on barrier membranes indicated that the collagen-based membranes appeared to facilitate cell attachment, whereas the polylactic acid-based membrane exhibited a morphology that was not conducive to attachment of human gingival fibroblasts. Based on these limited in vitro results, it appears that collagen-based membranes offer greater potential than polylactic acid-based membranes for guided tissue regeneration at surgical sites. PMID- 10453128 TI - Releasing severe adhesions around the eminence and the synovial portion of the TMJ: a clinical study of combined treatment using hydraulic lavage, arthroscopic surgery and rehabilitative therapy. AB - This paper evaluates the effects of treatment with a pumping technique and arthroscopic lysis and lavage, followed by rehabilitative training, on condylar head mobility of the temporomandibular joint (TMJ). We studied 32 TMJs in 19 patients suffering from chronic closed lock with severe adhesion. The results were compared between cases with adhesions concentrated in two areas: mostly in the posterior and/or the anterior synovial portion of the upper TMJ compartment (11 joints) and mostly around the eminence (21 joints). The results showed a statistically significant improvement in condylar head movement for both groups between the initial and final stages of treatment. However, the results also suggested that patients with adhesion concentrated around the eminence are less likely to recover condylar head mobility to the same extent as those in the other group. PMID- 10453129 TI - Measurement of airborne radon concentrations at several sites in a radioactivity research laboratory. AB - Radon-222 is a natural, gaseous, radioactive nuclide released from the ground and building materials into the air. Radon and its daughter nuclides can be an important disturbance factor for the measurement of environmental radioactivity. Radon concentrations in air in a radiation laboratory were measured with PICO-RAD detectors, which directly adsorb radon gas on activated charcoal. Generally, radon concentration increased in the absence of ventilation; a high concentration was observed in a radioisotope storage room without ventilation. Concentrations were low in other rooms used for experiments and measurement, which suggests that the radiation control practice in this laboratory is satisfactory and that the influence of natural radon gas on the measurement of radioactivity is negligible. PMID- 10453130 TI - Caries inhibiting and remineralizing effect of xylitol in vitro. AB - This study aimed to determine the effect of xylitol on the development and remineralization of caries in vitro, and to compare this effect with that of fluoride alone and in combination. Two experiments were devised. In experiment 1, bovine incisors were each sectioned into 4 portions which were randomly assigned to 4 demineralizing agents: A) acidic buffer (x), B) x + 0.5 ppm fluoride, C) x + 20% xylitol, and D) x + 20% xylitol + 0.5 ppm fluoride. Caries-like lesions were produced in specimens. In experiment 2, carious lesions were produced in teeth. Five lesion-bearing slabs were cut from each tooth. While one was reserved as control (UN), others were randomly assigned to 4 remineralizing agents: 1) artificial saliva (y), 2) y + 0.05 ppm fluoride, (3) y + 20% xylitol, and 4) y + 20% xylitol + 0.05 ppm fluoride. Mineral loss (delta Z) and lesion depth (ld) were quantified after 4-week remineralization. In experiment 1, numerical values of delta Z and ld observed can be ranked as A > C > B > D. These differences were significant only in B and D when compared with A for delta Z, but not between any group for ld. In experiment 2, the numerical values of delta Z and ld for control UN (unremineralized) and remineralized groups (1-4) ranked as UN > 3 > 4 > 1 > 2. Compared with UN, this difference was significant in all groups with ld, but not in any group with delta Z. We concluded that tolerable levels of xylitol alone may not show a significant caries inhibiting and remineralizing effect, but may act as a caries inhibitor additively with fluoride. PMID- 10453131 TI - Effect of calcitonin on bone healing following titanium implant insertion. AB - The purpose of this study was to evaluate the effects of calcitonin administration on bone healing, following titanium implant insertion. Thirty adult New Zealand rabbits received one implant in each femur. Fifteen animals were randomized to provide test (calcitonin) and control (saline solution) groups. The animals were sacrificed 7, 14, 21, 28 and 42 days after the surgical procedure. The analyzed parameters were new endosteal/periosteal bone length, new endosteal/periosteal bone area and total cortical length. Considering treatment, histometric measurements showed significant differences (p < 0.05) favoring the control group considering periosteal bone length and periosteal bone area, the other parameters were statistically similar between control and test groups. PMID- 10453132 TI - Statistical analysis of salivary pH changes after the intake of black tea and yerba mate supplemented with sweeteners. AB - Many studies have demonstrated the effect of various beverages on hard tissues of the mouth. Sugar consumption has also been avoided because it is associated with caries activity, and artificial sweetener research has been promoted. In this paper, data about the buffer capacity of black tea and yerba mate supplemented with sugar or one of two artificial sweeteners (Barny diet and NutraSweet) were examined. Salivary pH variations after the ingestion of both infusions were statistically analysed. A factorial design of four variables (Infusion, Sweetener, Concentration and Time) and their effects were considered independently and combined. The addition of sucrose lowered the pH of the infusions while black tea showed a greater buffer capacity than yerba mate. Significant differences (p < 0.0001) were obtained when salivary pH values were compared between infusions supplemented with sugar or artificial sweeteners. When a factorial design was used to examine differences between Barny diet and NutraSweet, no significant two- or three-way interactions could be observed. This statistical analysis showed a small significant variation in salivary pH after the ingestion of black tea or yerba mate with sugar but not with artificial sweeteners at the recommended concentrations. PMID- 10453134 TI - So-called simple bone cyst of the jaw: a family of pseudocysts of diverse nature and etiology. AB - The nature and etiology of so-called simple bone cyst (SBC) are still a subject of debate. Our comprehensive review of the literature suggests that SBC, which appears histologically to be a single entity, has different natures and etiologies, resulting in divergent clinical features. In addition, an interesting case of mandibular SBC in an 11-year-old girl is presented with details of radiographic changes over a 7-year period. Fully documented patient records revealed that this lesion originated in the apical area of the first molar and took about 4 years to develop into a clinically evident bony expansion. PMID- 10453133 TI - Osseointegration around titanium screws placed into the areas between guided bone augmented sites compared with osseointegration around guided bone graft augmented sites in rabbit tibia. AB - The aim of this study was to compare the degree of osseointegration around titanium screws placed into the areas between guided bone augmented sites and guided bone graft augmented sites in 8 Japanese white rabbit tibia. The skin and periosteum were incised and lifted to expose the inner surfaces of both tibia. Eight rabbits were divided into 2 groups: 1) a guided bone augmentation (GBA) group of 2 rabbits, and 2) a guided bone graft augmentation (GBCA) group of 6 rabbits. In the GBA group, the bone marrow was penetrated several times in both tibial bones. Three titanium screws were inserted into the bone so that the top of each screw was 3 mm above the bone surface and covered with an expanded polytetrafluoroethylene membrane (ePTFE). In the GBGA group autogenous cortical bone (10 mm x 10 mm x 1 mm) was taken from the left tibia and grafted on the inner surface of the right tibial bone, titanium screws being similarly inserted. The surgical site was covered by ePTFE. After healing for 2 months, the animals were euthanatized, and the experimental area was prepared for histological investigation. New bone had formed on the surgical sites in the GBA group and grafted bone had survived in the GBGA group, with no sign of infection or membrane exposure. The degree of osseointegration around the screw was 67.6 +/- 11.2% (mean +/- SD) in the GBA group and 78.8 +/- 11.9% in the GBGA group. There was no significant difference between the groups (p = 0.29). The ratio of mineralized bone to bone marrow was 45.7 +/- 6.2% in the GBA group and 64.4 +/- 4.4% in the GBGA group. There was a significant difference between the groups (p < 0.05). Although there was no significant difference for osseointegration around the screw placed into the GBA and GBGA sites, the quality of newly generated bone or grafted bone was quite different. Therefore, we suggest that predictable bone augmentation beyond the bone surface can be achieved by either the GBA or the GBGA procedure with ePTFE. However, it was difficult to conclude which procedure was superior with respect to the osseointegration around the titanium screw. PMID- 10453135 TI - Teenage pregnancy. PMID- 10453136 TI - Arkansas women and osteoporosis: informed but still at risk. PMID- 10453137 TI - The accuracy of non-bronchoscopic bronchoalveolar lavage in the diagnosis of pneumocystis carinii Pneumonia in children. PMID- 10453139 TI - Hill's sign: a non-invasive clue of the severity of chronic aortic regurgitation. PMID- 10453140 TI - What's new in the public health laboratories? PMID- 10453141 TI - The Mason Brown Lecture. Scots and paediatric surgery. AB - Scotland and Scots paediatric surgeons have been in the forefront of the development of surgical services for children. Development has occurred at different rates in sections of society. Progress or obstruction has seldom been for 'scientific' reasons but much more according to the personal interests or drive of individuals. The Scots can justifiably claim some influence in the overall evolution of paediatric surgery. It is no wonder that Henley wrote after his visit to Walter Scott in Edinburgh, "The conceit of these bloody Scotsmen is atrocious". PMID- 10453142 TI - Cardiovascular surgery in the elderly. AB - Throughout the developed world, the population is aging at an alarming rate. This has become an issue of great concern. The growing number of elderly people within the community has several health and economic implications. The prevalence of atherosclerosis increases with age and directly impacts on the workload of both cardiac and vascular surgeons. The decision to operate should not be based on age alone but reflect an assessment of the risk benefit ratio of individual cases. Increased life expectancy, safer anaesthesia and less invasive surgical techniques have led to more patients being considered for surgical intervention. However, this will inevitably result in an overwhelming burden on national health resources, a factor that may force some practitioners to deny treatment on the grounds of chronological and not biological age. PMID- 10453143 TI - Dieulafoy's lesion: an obscure cause of GI bleeding. AB - Dieulafoy's lesion is an uncommon cause of major gastrointestinal bleeding and may be difficult to recognise. It consists of an arteriole that protrudes through a tiny mucosal defect, usually within 6 cm of the gastroesophageal junction on the lesser curve of the stomach. Similar lesions have also been described in the distal oesophagus, small intestine, colon, and rectum. Awareness of the condition and experience in endoscopy are the mainstay of diagnosis. Therapeutic endoscopy is the first line of treatment. It is safe, effective and has very good long term results. PMID- 10453144 TI - The accuracy of "one-stop" diagnosis for 1,110 patients presenting to a symptomatic breast clinic. AB - To minimise delay in diagnosis and reduce patient anxiety, triple assessment with immediate reporting has been used in our symptomatic breast clinic since 1991. This article examines the accuracy of the diagnostic modalities used and the efficacy of the "one-stop" diagnostic policy. The data on 1,110 new patients presenting to the symptomatic breast clinic between January and July 1993, were analysed and subsequent three year follow-up and outcome established. Fine needle aspiration cytology (FNAC) gave the highest predictive value (97.3%) with a sensitivity of 93.5% and a specificity of 98.1%. Ultrasonography provided a 97.0% prediction with a sensitivity of 88.9% and a specificity of 97.4%, whereas mammography had a prediction of 96.4% with sensitivity of 93.2% and a specificity of 96.7%. When the mammogram or ultrasound scan were reported as unequivocally benign (M1), there were no missed cancers. The false positive and false negative rates for FNAC were 0% and 1.4%, respectively. Following assessment, a diagnosis was made in 96% of patients. Sixty-two percent of the patients were discharged at the first clinic visit. Four breast malignancies were subsequently diagnosed in the discharged group; two with new microcalcifications due to ductal carcinoma in situ, one with invasive disease in a different quadrant of the breast from that originally investigated, and in one patient the cancer was missed. CONCLUSION: A "one-stop" symptomatic breast clinic provides an accurate and effective means of establishing a correct diagnosis. PMID- 10453145 TI - Effect of curative versus palliative surgical treatment for stage III pancreatic cancer patients. AB - During the last decade, significant progress has been made in pancreaticoduodenectomy for patients with pancreatic carcinoma. Pancreatic resection performed by surgeons in tertiary referral centres is therefore justified, while the indications for pancreatic resection could be extended in patients with advance stages of disease. The aim of our study is to compare the effect of curative (pancreaticoduodenectomy) versus palliative surgery in patients with stage III pancreatic cancer, during a 20-years period. We retrospectively reviewed the charts of 58 consecutive patients with stage III ductal adenocarcinoma of the head of the pancreas. 23 patients underwent pancreatoduodenectomy with curative intent while the remaining 35 patients had surgery for palliative purposes (combined biliary and gastric bypass was performed in 83%). The hospital mortality rate was similar in both groups (4% vs 6%). 43% of patients undergoing pancreaticoduodenectomy had an uncomplicated post operative course compared with 49% of patients undergoing palliative bypass. The length of surgical procedure and post-operative hospital stay in pancreaticoduodenectomy group were significant longer compared to those patients undergoing palliative bypass (p = 0.03 and p = 0.02 respectively). The overall actuarial survival was significantly (p < 0.01) longer in the group of patients who underwent pancreaticoduodenectomy compared with the group with palliative intent surgery. CONCLUSION: Pancreaticoduodenectomy with curative intent for stage III pancreatic cancer patients, could improve prognosis with similar peri operative morbidity and mortality when compared with palliative bypass. PMID- 10453146 TI - Tendon allografts in knee ligament surgery. AB - This paper describes the experiences of the Orthopaedic Service and the Scottish National Blood Transfusion Service in Edinburgh in establishing a tissue bank for allograft tendons used in knee ligament reconstruction surgery. Ninety-five tendons have been harvested from 23 donors, and 36 grafts have been implanted in 30 knees for either primary ligament reconstruction (group 1, 4 knees), revision ligament reconstruction (group 2, 11 knees), or in patients with multiple ligament deficiency (group 3, 15 knees). The basic science of ligament allografts, donor screening and the role of tendon allografts are reviewed. PMID- 10453147 TI - Standardisation of radiographic views of the fractured tibia. AB - Serial radiographs are difficult to compare when the position of the shin is not accurately controlled in rotation. We have designed a device which can be used easily by the radiographer to produce accurately standardised radiographs of the tibia. PMID- 10453148 TI - Open 'Bankart' repair for recurrent anterior dislocation of the shoulder. AB - "... the frequently recurring dislocation from trivial causes is a great and serious disability" (AS Blundell Bankart, 1923). PMID- 10453149 TI - Measurement of quality of life in surgery. AB - Quality of life has become an important outcome measure in surgery. Its use as an end-point in clinical trials is well recognised. However, quality of life measurement in surgery is currently done mainly for research purposes, and still has to find a place in routine clinical practice. This paper presents an overview of the methodology and uses of quality of life assessments in surgical practice. PMID- 10453150 TI - Auditing outcomes after treatment of Scottish patients with uveal melanoma in Liverpool. AB - Uveal melanoma is the most common primary intraocular malignant tumour, although it is a rare condition. Between January 1993 and December 1997 a total of 120 patients were referred from Scotland to the Ocular Oncology Service at St. Paul's Eye Unit in Liverpool. Of these, 98 had uveal melanoma, which was treated with primary enucleation (24), or by conservative methods, consisting of plaque radiotherapy (30), proton beam radiotherapy (19), trans-scleral local resection (19), or endoresection (6). After conservative treatment, 92% of eyes were retained, with 65% of eyes maintaining the same level of vision as at presentation or better. The main complications were retinal detachment (8), death from metastatic disease (7) and local tumour recurrence (4). This study gives an overview of the treatment of uveal melanoma and demonstrates that most patients with uveal melanoma can be treated conservatively, with the large majority retaining the eye and good vision. PMID- 10453151 TI - The quality of surgical information on the Internet. AB - The quality of surgical information on the Internet is variable. Content variation limits the use of the Internet as a reliable and safe information source both for both health professionals and patients. Medical information is a particularly sensitive area. Incorrect or misleading information may lead to potentially dangerous health behaviour, patients reading information intended for health professionals may misunderstand information or may get wrong expectations regarding treatment options. PMID- 10453152 TI - Ten essential clinical informatics skills. AB - Information skills are basic to good medical practice. Every clinician needs to understand the principles of data interpretation, the logical foundations of the diagnostic process, and the management of uncertainty in clinical knowledge. Understanding the implications of using one medium rather than another is central to developing effective communication skills as these communication options become commonplace in the community. PMID- 10453153 TI - Pancreatic pseudocyst: heparin-induced haemorrhage through the ampulla of Vater. AB - Bleeding from a pancreatic pseudocyst through the ampulla of Vater is a rare cause of overt gastrointestinal haemorrhage. Previously described mechanisms of such haemorrhage are reviewed. We report a case of a 74-year-old female with a pancreatic pseudocyst that bled into the gastrointestinal tract following the administration of heparin. We believe that this is the first reported case of its kind. PMID- 10453154 TI - Haemosuccus pancreaticus due to mucinous cystadenocarcinoma: the significance of recurrent abdominal pain, hyperamylasaemia and a pancreatic cyst in association with recurrent gastrointestinal bleeding. AB - Haemosuccus pancreaticus is a rare cause of gastrointestinal haemorrhage, and when it presents in otherwise healthy people, can prove difficult to diagnose. The cardinal features are episodic epigastric pain associated with a raised serum amylase and the passage of melaena. Failure to make the connection between recurrent gastrointestinal bleeding and apparently unrelated symptoms attributable to pancreatitis may lead to a significant delay in diagnosis. PMID- 10453155 TI - The management of femoral and tibial diaphyseal fractures. 1998;43: 374-80. PMID- 10453156 TI - Local anaesthesia: to warm or not to warm? A survey of current practice. 1999;44:167-71. PMID- 10453157 TI - Re: How to cope with unsolicited email from the general public seeking medical advice. PMID- 10453158 TI - [Clinical results of surgical repair for thoracic aortic aneurysms: intraoperative blood loss and morbidity]. AB - Clinical results of 64 patients who underwent surgical repair for thoracic aortic aneurysms were studied, focusing on the relationship between intraoperative blood loss and postoperative morbidity. Operative mortality was 22% in the urgent repair group and 9% in the elective repair group. Deep hypothermia, operative death, postoperative complication were the factors which significantly correlated to the amount of intraoperative blood loss. In patients who received deep hypothermia, larger blood loss and higher incidence of mortality and morbidity were observed. There was a significant relationship between the lowest core temperature during cardiopulmonary bypass and the lowest platelet count. Intraoperative blood loss revealed as a strong risk factor to directly influence postoperative clinical outcome of thoracic aortic operation. PMID- 10453159 TI - [A case report of concomitant operation of minimally invasive direct coronary artery bypass and left lower lobectomy]. AB - A 72-year-old man was admitted complaining of laryngeal discomfort. A coronary angiogram showed a total occlusion of the LAD, a 90% stenosis of the 1st diagonal branch and well-developed collaterals from the LCX to the LAD. A chest X-ray showed a consolidation with unclear border at the left hilus of the lung. A lung biopsy through the bronchus revealed an adenocarcinoma of the left lower lung. He underwent a concomitant CABG operation without a cardiopulmonary bypass and a left lower lobectomy with lymph node dissection. The CABG was performed by left anterior small thoracotomy in the supine position and the lobectomy was performed by left posterolateral thoracotomy in the right lateral position. An intraoperative postural conversion was useful to obtain a good field in each operation and to prevent hemodynamic deterioration due to heart compression. PMID- 10453160 TI - [Dumbbell type schwannoma of the posterior mediastinum: a report of two cases with different surgical approaches]. AB - Two cases of dumbbell type schwannoma of the posterior mediastinum are reported. Case 1 involved a 16-year-old man and case 2, a 48-year-old woman. They were asymptomatic, and in both cases an abnormal mass shadow of the left mediastinum was incidentally detected on a routine chest roentogenogram. In these cases, CT and MRI revealed that dumbbell type tumors extended to the spinal canal. The operations were performed with thoracic approach cooperated with a neurosurgeon in case 1 and with combined posterior and thoracic approach cooperated with an orthopedic surgeon in case 2. Histopathologic findings indicated all benign schwannomas. The reported 25 cases in Japan sofar are reviewed. PMID- 10453161 TI - [Clinical experience with gelatin-resorcine-formol glue for repair of postinfarction ventricular septal rupture]. AB - We report a surgical technique for treatment of postinfarction ventricular septal rupture (VSR). An 82-year-old woman underwent successful surgical repair of a VSR two days after suffering anterior myocardial infarction. After repair of the VSR with a Dacron patch, the left ventriculotomy was closed with mattress sutures over felt strips using the gelatin-resorcine-formol glue to reinforce the left ventricular wall. The patient was doing well four months after surgery. PMID- 10453162 TI - [Efficacy of mupirocin in eradicating methicillin-resistant Staphylococcus aureus from nasal discharge in carrying cardiovascular surgical patients]. AB - Methicillin-resistant Staphylococcus aureus from nasal discharge was identified in 37 (2.5%) cardiovascular patients operated between 1995 and 1997; 25 male and 12 female, ranging from 1 to 83 years (mean 63); 2 were excluded because of Arbekacin or Isodine-gel treatment. The first 17 were treated with Vancomycin inhalation (V group) and eradication was considered to have been achieved when 3 consecutive negative cultures were obtained; the subsequent 18 were treated with Mupirocin (M group) and eradication was determined by 1 negative culture. In post eradication electively operated 13 V and 15 M, postoperative MRSA infection was observed in one M (wound infection); the interval from the first nasal culture to the operation was 68 +/- 58 in V and 32 +/- 12 days in M, respectively (p < 0.05). In the remaining 7 who had to undergo emergency surgery while waiting for eradication because of progression of symptoms (2 V) or prior to instituting treatment (2 V, and 3 M), postoperative MRSA infection was observed in 2 M (both pneumonia). No deaths from infection were observed. Though the time required for conversion of the nasal culture was longer in V (13 +/- 20) than in M (7 +/- 1 days) differences were not significant. Mupirocin is easier to use, eradication can be achieved generally within a week. PMID- 10453163 TI - [Lung resection affects the postoperative arrhythmia and electrocardiographic axis deviation]. AB - After lung lobectomy or pneumonectomy, the mediastinal shift and diaphragmatic elevation are occurred. Because this phenomenon may affect the heart positional change, we studied the electrocardiographic QRS axis in the frontal plane (from leads I and III) and the postoperative arrhythmia. Seventy three patients who had no heart disease including arrhythmia before the surgery were recorded their electrocardiogram (ECG) before their surgery and after their discharge. When the postoperative ECG was recorded, they had no respiratory failure nor cancer recurrence, and their lungs were fully expanded in their thoracic cages. After right upper lobectomy (19 cases), the axis was twisted rightward slightly (2.1 degrees). Right middle lobectomy (2 cases, 9.5 degrees) and right upper and middle lobectomies (3 cases, 7.3 degrees) twisted the heart axes more rightwards. Right lower lobectomy (12 cases, -1.0 degree) and right middle and lower lobectomies (3 cases, -17.7 degrees) contorted their axes leftwards and right pneumonectomy (5 cases, 31.4 degrees) rightwards. The axes were turned rightwards after the left upper lobectomy (18 cases, 2.8 degrees) and the left lower lobectomy (7 cases, 3.9 degrees). Left pneumonectomy (4 cases, -4.0 degrees) twisted the axis leftwards. After the surgery, arrhythmias were recorded in 14 cases and, among these patients, 5 cases were required the oral anti-arrhythmic medication. Most of these cases changed their heart axes after the surgery and it is suggested that the axial deviation may contribute to their postoperative arrhythmia. PMID- 10453164 TI - [The analysis for surgical procedures of coronary-artery bypass grafting: selection of graft materials]. AB - We analyzed 222 patients undergoing coronary artery bypass grafting (CABG) in our institute. Our selection of graft materials consists of only one arterial conduit and one or more saphenous vein grafts (SVG). An arterial conduits (left internal thoracic artery (LITA) was mainly used for the left anterior descending coronary artery (LAD), while a SVG was used for coronary arterial branch. Our approach was evaluated by the mid-term prognosis as well as cardiac events. Seventy-one percent of patients with CABG could be followed. The graft patency rate was better for ITA grafts than for SVG (97.8% vs 88%). The incidence of late cardiac events was lower in patients with the LITA, to the LAD. Furthermore, these patients had a better survival rate at 4 years comparing with patients who had vein bypass grafts alone. We suggest that this selection of graft materials may be accepted in CABG for the good quality, avoiding the cardiac events. PMID- 10453165 TI - [Predonation and transfusion in open heart surgery]. AB - The efficacy of predonation of autologous blood in reducing the use homologous blood during open heart surgery was investigated. Between January 1997, and February 1998, predonation and transfusion was studied in 100 consecutive open heart operations (CABG, 77; valve surgery, 17; ASD, 5; myxoma, 1). The guidelines for autologous predonation were as follows: an age < 70 years, a weight > 40 kg and a hemoglobin > 12 g/dl. Patients in NYHA class IV or undergoing emergency operation were excluded. The blood loss during operation ranged from 195 to 1,850 ml (mean; 670 ml), being from 305 to 1,850 ml (723 ml) for CABG, from 260 to 1,020 ml (493.5 ml) for valve surgery and from 195 to 570 ml (342 ml) for ASD. The blood loss was not significantly dependent on sex or age and did not differ elective and emergent operations. Only 36.6% of patients with autologous predonation needed homologous transfusion versus 63.4% of those without predonation. Homologous transfusion was done in only 5% of the those with predonation of 800 ml versus 69% at 400 ml and 71% at 200 ml. In conclusion, autologous blood transfusion is effective for reducing the homologous blood requirement. It also seems that predonation of 800 ml may be sufficient to allow open heart surgery without blood transfusion. PMID- 10453166 TI - [A report of 4 cases of aortic operation using open proximal anastomosis under the hypothermic circulatory arrest and left lateral exposure]. AB - Four cases of aortic operations using open proximal anastomosis (OPA) under the hypothermic circulatory arrest (HCA) and left lateral approach (LLA) are reported. Three of 4 cases had extensive aortic disease from distal arch to descending thoracic aorta (Stanford type B chronic aortic dissection, double false aneurysms, and double true aneurysms). Another one case had ruptured aneurysm of thoracic aorta. LLA should have been selected in all cases, however, aortic proximal cross-clamp was impossible in them, because of giant pseudolumen, diseased lesion of aortic arch, hemothorax followed rupture of aneurysm. Therefore OPA under the HCA was performed. There were no complication associated with HCA, bleeding, neurological deficiency and respiratory dysfunction. We conclude that, although the HCA may have some problems, if there is the proper indication, OPA under the HCA is useful method at aortic operation for difficult aortic disease. PMID- 10453167 TI - [Two cases of coronary artery bypass grafting associated with antiphospholipid syndrome in systemic lupus erythematosus]. AB - We report two cases of coronary artery bypass grafting (CABG) associated with antiphospholipid syndrome (APS) in systemic lupus erythematosus (SLE). Patient 1, 65-year-old female, who had been treated for SLE with prednisolone for 11 years was transferred to our hospital due to unstable angina caused by stenosis of the left main trunk (LMT) and the left anterior descending artery (LAD). She underwent emergency CABG of the LAD using left internal thoracic artery (LITA). Post operative doppler study demonstrated patent LITA to the LAD. Patient 2, 67 year-old female who had been treated for SLE with prednisolone for 8 years was transferred to our hospital due to acute myocardial infarction caused by stenosis of the LMT and the left circumflex artery (LCX). She underwent emergency CABG of the LAD and the LCX using saphenous vein grafts (SVGs). Post operative angiography confirmed a patent SVG to the LAD and an occuluded SVG to the LCX. In cases of SLE, the frequency of occurrences of ischemic heart diseases is high. Until now, however, there are few instances reported on performing CABG for patients with SLE. We are reporting here our particular cases of APS with SLE, discussing the involvement of APS as causative factor of ischemic heart diseases and related issue of surgical and post surgical antithrombotic treatments. PMID- 10453168 TI - [A case report of surgical repair of ruptured aneurysm of sinus valsalva in aged (84 years old)]. AB - An 84-year-old man with ruptured aneurysm of Valsalva sinus was operated. Diagnosis was made by two dimensional echocardiography, cardiac catheterization, and cardiac angiography. A aneurysm was found at the right-coronary sinus region, and ruptured into the right ventricle. The ruptured aneurysm of sinus Valsalva was repaired with direct closure and Woven patch from inside the right ventricle through the pulmonary valve. This case was defined as congenital because there was no sign of inflammatory or atherosclerotic changes in the aorta, aortic valve and aneurysm. The ruptured aneurysm of sinus Valsalva is very rare in aged patients. As far as we know, this patient is one of the oldest cases who underwent successful surgical repair in this country. PMID- 10453169 TI - [Mediastinoscopic resection combined with a sternum lift technique in a case of thymic cyst]. AB - We report on a case of thymic cyst which was successfully treated with mediastinoscopic resection combined with a sternum lift technique. A 62-year-old man was admitted to our hospital with a thymic cyst, which was causing chest discomfort and was increasing in size. The tumor was located above the pulmonary artery in the anterior mediastinum. He underwent mediastinoscopic resection while lifting the inferior portion of the sternum with a Laparolift. The cyst was resected sharply and bluntly with the Harmonic Scalpel without rupture. The patient was discharged on the 6th postoperative day in a good condition. Mediastinoscopic resection of a thymic cyst through a xiphoid approach is a useful surgical approach because of the absence of postoperative pain and because it allows early discharge from the hospital. PMID- 10453170 TI - [A case of solitary metastatic lung tumor from follicular carcinoma of the thyroid in hemodialysis patient]. AB - A 61-year-old female was admitted for abnormal shadow of chest X-ray film. The patient has been treated with hemodialysis for chronic renal failure since February 16, 1982, and was performed total thyroidectomy for follicular carcinoma of thyroid on October 24, 1989. Chest X-ray on September 8, 1994 showed the abnormal shadow of right lower lung, chest CT revealed tumor shadow which is suspected to malignancy. We performed partial resection of right lung including tumor on October 19, 1994. Histological examination certified the lung metastasis of follicular carcinoma of thyroid. Postoperative course has been uneventful and there has been no episode of recurrence for 3 years and half after the surgery. We reported this very few case with solitary metastatic lung tumor from follicular carcinoma of thyroid with a review of the literature. PMID- 10453171 TI - [A case of thoracoscopic resection of a pulmonary infarction mimicking a metastatic lung tumor]. AB - We report here a rare case of pulmonary infarction which was resected as a metastatic pulmonary tumor. A 57-year-old male patient referred for exhibiting an abnormal shadow on chest X-ray 8 months after the resection of invasive adrenal tumor. The patient underwent thoracoscopic resection of the lesion as a metastatic tumor. However, intraoperative pathological examination revealed that the resected specimen was pulmonary infarction. Post-operative MRI and ultrasound images showed no evidence of venous thrombus. Pulmonary infarction can exhibit an abnormal shadow in the lung fields, and it is necessary to be distinguished from true tumor. PMID- 10453172 TI - [A case of surgical treatment for thymic Hodgkin's disease with tracheo-bronchial stenosis]. AB - An 18-year-old female was introduced our hospital for facial swelling and subcutaneous tumor of anterior neck. Chest X-ray and CT films showed mediastinal mass with tracheo-bronchial stenosis. Incisional biopsy specimens of neck tumor revealed that the tumor was suggestive of malignant lymphoma. Because of progressive dyspnea after CHOP regimens the operation was performed and histopathological examinations revealed thymic Hodgkin's disease. After operation dyspnea was improved but dysphagia and hoarseness occurred. These symptoms were getting improved. She received 6 cycles of ABVD regimens and discharged after 5 months. PMID- 10453174 TI - Dietary carotenoids and certain cancers, heart disease, and age-related macular degeneration: a review of recent research. AB - Key epidemiologic studies show associations between high dietary intakes of certain carotenoid-containing fruits and vegetables and reduced risk of prostate cancer, breast cancer, head and neck cancers, cardiovascular disease, and age related macular degeneration, although overall the evidence is inconsistent. Little is known about the potential biochemical mechanisms whereby carotenoids might protect against disease, and human intervention trials are limited to high dose beta-carotene, which is not protective against lung cancer or cardiovascular disease. Authoritative scientific organizations continue to emphasize increased consumption of fruits and vegetables but do not make specific recommendations for carotenoids because of a lack of data that directly link them to disease reduction. PMID- 10453173 TI - [A case of metachronous lung cancer in which tumor developed within the different ipsilateral lobe over a short period of time and completion pneumonectomy was required]. AB - A 51-year-old man underwent a middle-lower lobectomy for squamous cell carcinoma on February 8, 1996. In July, 1997, a computed tomography revealed a mass shadow in the right upper lung field. Completion pneumonectomy was performed. Histopathological examination showed poorly differentiated adenocarcinoma. This case was the shortest time to occurrence of second tumor in our metachronous lung cancer cases. We must always give attention to exist second primary lung cancer and double primary lung cancer after resection of primary lung cancer. PMID- 10453175 TI - Cigarette use during adolescence: effects on nutritional status. AB - Despite numerous anti-tobacco campaigns, smoking among adolescents continues to be a predominant public health issue. This report details the interrelationships between adolescence, smoking, and nutrition and health. Current data indicate that most smokers become nicotine-dependent as adolescents, which places them at risk for chronic diseases associated with continuous oxidative damage. Additionally, nicotine has antidepressant and hypermetabolic effects, which may be of particular importance during adolescence because nicotine use leads to increased dependence on tobacco, contributes to difficulty in smoking cessation, and promotes weight gain following smoking cessation. PMID- 10453176 TI - Criteria and recommendations for vitamin C intake. AB - The experimental evidence that forms the basis for setting recommended intake levels of vitamin C for humans is currently undergoing review. A recent study suggests that a vitamin C intake of 100 mg-200 mg/day is needed, which is two to threefold greater than the current Recommended Dietary Allowance. This study also proposes that individuals consume no more than 1 gram of vitamin C per day. PMID- 10453177 TI - Food-borne antibiotic-resistant Campylobacter infections. AB - A recent report has provided some important information concerning the apparent emergence of quinolone-resistant bacterial infections in the United States. In particular, this report indicates an increase in domestically acquired, quinolone resistant Campylobacter jejuni infections, likely associated with poultry consumption. PMID- 10453178 TI - Magnesium supplementation and bone turnover. AB - Two-thirds of total body magnesium content is located in the skeleton. Recently, there have been reports that high dietary magnesium intakes are associated with higher bone mineral density. The effect of magnesium supplementation on bone turnover has recently been investigated in young adults. However, the findings from these studies are conflicting. More studies are necessary to better elucidate the role of magnesium in bone health. PMID- 10453179 TI - The girl child and her equal right to 'first call' for children. AB - Using the life-cycle approach, the problems of the girl child are summarized. A number of recommendations are put forward. Also enumerated are tasks that child health care providers can focus on to address these problems. Pertinent United Nations initiatives for the protection of women/girls are listed, in addition to the output of a pre-Congress workshop on the girl child sponsored by the International Pediatric Association, the World Health Organization and UNICEF. PMID- 10453180 TI - Analysis of tumor necrosis factor-alpha production and polymorphisms of the tumor necrosis factor-alpha gene in individuals with a history of Kawasaki disease. AB - BACKGROUND: Tumor necrosis factor (TNF)-alpha plays a central role in the pathogenesis of vasculitis in Kawasaki disease (KD). To address the genetic background of KD, we investigated the level of TNF-alpha production and genetic polymorphisms in the 5' flanking region of the TNF-alpha gene in healthy children with a history of KD. METHODS: For TNF-alpha production, peripheral blood mononuclear cells (PBMC) of children with a history of KD (n = 61) and of non-KD children (n = 35) were stimulated with phorbol 12-myristate 13-acetate, toxic shock syndrome toxin-1 (TSST-1) and the culture supernatant of Staphylococcus aureus derived from a KD patient (S-6), which had several superantigenic activities. The genetic background of KD was addressed by studying polymorphisms in the 5' flanking region of the TNF-alpha gene at positions -1031 (thymine (T) to cytosine (C) change, termed -1031C), -863 (C to adenine (A), -863A), -857 (C to T, -857T), -308 (guanine (G) to A, -308A) and -238 (G to A, -238A) in KD, using dot-blot hybridization with sequence-specific oligonucleotide probes. RESULTS: The PBMC of KD patients with coronary artery lesions produced slightly higher levels of TNF-alpha in response to the bacterial products (such as TSST-1 and S-6). None of the polymorphisms in the 5' flanking region of the TNF-alpha gene were related to KD. CONCLUSIONS: These results suggest that a genetic disposition towards overproduction of TNF-alpha in response to bacterial products may be involved in the pathogenesis of KD. PMID- 10453181 TI - Microsatellite analysis of childhood leukemia: correlation of 9p and 12p chromosome abnormalities with expression of related genes. AB - BACKGROUND: Loss of heterozygosity (LOH) and DNA replication error (RER) have been thought to be involved in carcinogenesis, but have not been investigated in childhood leukemia and lymphoma. METHODS: Eighty samples from 65 patients with childhood leukemia and lymphoma were examined using seven different microsatellite markers for RER analysis. Additionally, LOH in two chromosome regions (9p and 12p) was investigated. Furthermore, expression of the TEL, TEL/AML1 and p27(KIP1) genes on 12p and the p16 gene on 9p were detected by reverse transcriptase polymerase chain reaction. RESULTS: Replication errors were detected in 5/65 patients (7.7%). Most (4/5 patients) RER were preferentially located in the 9p and 12p regions. There were two patients who had DNA abnormalities in both 9p and 12p, one with common acute lymphoblastic leukemia (ALL) showed 9p LOH and the TEL/AML1 fusion gene on 12p and the other with common ALL and 12p RER had diminished expression of both the p27(KIP1) gene on 12p and the p16 gene on 9p. CONCLUSIONS: Combined DNA alterations on 9p and 12p, involving LOH, RER and/or gene mutation and chromosomal translocation, were found in childhood acute leukemia, especially in common ALL. PMID- 10453182 TI - Clinicopathologic correlations of Henoch-Schonlein nephritis in Turkish children. AB - BACKGROUND: In this study, 66 patients with Henoch-Schonlein nephritis (HSN) were investigated clinicopathologically. METHODS: The patients were classified according to their initial presentation, histologic findings, recurrences of purpura, type of treatment and clinical outcome. Logistic regression analysis was performed. RESULTS: Sixty-eight percent of patients were hospitalized with mild renal disease. Most patients were evaluated as class I and II according to light microscopy. In addition to IgA deposition alone, 33% of patients showed IgA + C3 and 27% had IgA + IgG + C3 depositions. After the follow-up period of 3.3 years, 15 patients had minor urinary findings, 4 had active renal disease and 1 had renal insufficiency. Recurrences occurred in 37.9% of patients and 37.1% of patients with recurrences had persistent pathologic findings. Symptomatic treatment was given to 51.5% of patients, while 27.2% were given corticotherapy. CONCLUSIONS: Clinical presentation was found to be correlated with outcome. Recurrence of the disease and the type of the treatment also affected the outcome. It was also thought that mesangial IgG and C3 depositions may have a role in the pathogenesis of renal damage in HSN. PMID- 10453183 TI - Effects of iron deficiency anemia on hemoglobin A1c in type 1 diabetes mellitus. AB - BACKGROUND: The relationship between hemoglobin A1c (HbA1c) and iron status in type 1 diabetes mellitus (DM) has not been adequately studied. In this prospective investigation, we aimed to determine the effect of iron deficiency on HbA1c in diabetic patients who also had insufficient iron stores. METHODS: Thirty seven patients with type 1 DM were included in the study. Eleven of them were also iron deficient (ID) and the remaining 26 were iron-sufficient (IS). Two non diabetic control groups were selected for the ID and IS groups. All patients with ID were treated with iron at 6 mg/kg per day for 3 months. Glycemia in diabetic patients was monitored at home before breakfast and supper by a glycometer. Hemoglobin A1c was measured in all subjects at the beginning and the end of the study. RESULTS: Patients with ID DM had higher levels of HbA1c than those in the control group (P < 0.001). There were no significant differences in the weekly average glucose concentration of the patients with ID DM before and after iron supplementation. In contrast, HbA1c decreased from a mean of 10.1 +/- 2.7% to a mean of 8.2 +/- 3.1% (P < 0.05). Additionally, HbA1c in ID non-diabetic patients decreased from a mean of 7.6 +/- 2.6% to 6.2 +/- 1.4% after iron therapy (P < 0.05). CONCLUSIONS: We conclude that among type 1 DM patients with similar level of glycemia, iron deficiency anemia is associated with higher concentrations of HbA1c. In addition, iron replacement therapy leads to a drop in HbA1c in both diabetic and non-diabetic patients. The iron status of the patient must be considered during the interpretation of HbA1c concentrations in type 1 DM. PMID- 10453184 TI - Dopamine penetrates to the central nervous system in developing rats. AB - BACKGROUND: The purpose of this study was to evaluate whether or not dopamine (DA) can penetrate to the central nervous system (CNS) from the blood in the infantile period in rats. METHODS: In a preliminary experiment, we administered a 50 mg/kg dose of DA hydrochloride, intraperitoneally, to 7-day-old rats (DA 50 mg/kg group), obtaining cerebrospinal fluid (CSF) both before and at 5, 10, 20, 30, 60 and 120 min after administration. The CSF levels of DA and its main metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were then measured. Next, we investigated the DA transfer from blood to the CNS by administering doses of 1, 5, 10 and 30 mg/kg DA hydrochloride (DA 1, 5, 10 and 30 mg/kg groups). In these groups, CSF samples were obtained only at 10 and/or 60 min after DA administration, based on the results of the DA 50 mg/kg group. RESULTS: The DA concentrations in CSF significantly increased compared with values before DA administration in the DA 50 mg/kg group. The DA concentrations in the DA 30 mg/kg group, DOPAC concentrations in the DA 5, 10 and 30 mg/kg groups, and HVA concentrations in all groups were significantly higher than in the control (saline injection) group. CONCLUSIONS: These findings suggest easy DA transfer from blood to the CNS and immaturity of the blood-brain barrier for DA in the infantile period in rats. PMID- 10453185 TI - Shift in the buoyant density of hepatitis C virus particles in infants infected by mother-to-infant transmission. AB - BACKGROUND: Hepatitis C virus (HCV) particles in sera can be divided into two classes: low-density free particles and high-density immune complex particles. Previous studies have revealed that the clinical progression of HCV infection is closely associated with the occurrence of the former class, rather than the latter, in an experimental chimpanzee model and in HCV-infected adult cases. METHODS: To verify this concept in infantile cases, we prospectively analysed HCV particle populations, fractionated according to buoyant density, in serum samples from five infants infected by mother-to-infant transmission. RESULTS: In all five cases, HCV particles were predominantly high density at the age of one month. In four of five cases, low-density HCV particles became predominant in association with a decrease in maternally transmitted antibody levels. In one case, in which high serum levels of alanine aminotransferase persisted, low-density particles were predominant between the ages of 3 and 9 months, in three consecutive samples. In other cases, in which infants were asymptomatic or had transient hepatitis, low-density HCV particles were predominant at only one sampling point or not at all throughout the follow-up period. CONCLUSIONS: Maternal antibody transmitted via the placenta reacts with the HCV particles in infants infected through vertical transmission. A decrease in maternal antibody levels results in an increase in low-density free virions. It is suggested that low-density particles play an important role in liver inflammation. PMID- 10453186 TI - A mechanism for shock following stomach rupture in the newborn. AB - BACKGROUND: Stomach rupture often leads to shock and death within a short period, but the mechanism for this is not well-known. Shock may be due, in part, to endotoxin translocation and endotoxemia. METHODS: Sterile, endotoxin-free HCl (0.1 mol/L), with or without endotoxin-feeding, was injected intraperitoneally into 10-day-old rats, simulating stomach rupture in the newborn. The plasma endotoxin concentration was measured. Plasma glucose and lactate concentrations were monitored to assess physiologic response to endotoxemia and shock. RESULTS: Endotoxin feeding alone caused endotoxemia (0.49 +/- 0.12 ng/mL; P < 0.05) in 10 day-old rats, while plasma endotoxin concentration in controls was less than 0.04 ng/mL. However, the endotoxemia did not cause disruption of glucose regulation or death. Injection of HCl alone did not increase plasma endotoxin concentrations and did not alter glucose regulation. However, HCl injection into endotoxin-fed rats induced endotoxemia (211.1 +/- 70.5 ng/mL; P < 0.05), hypoglycemia, lactacidemia and mortality (45%; P < 0.05). CONCLUSION: Early development of shock following stomach rupture may be in part due to endotoxin translocation and endotoxemia. PMID- 10453187 TI - Contracted form of endocardial fibroelastosis in two siblings. PMID- 10453188 TI - Successful conservative management of chylopericardium after bilateral Blalock Taussig shunts. PMID- 10453189 TI - Fulminant ulcerative colitis associated with both masseter muscle myositis and immunoglobulin M nephropathy. PMID- 10453191 TI - A case of megalocornea-mental retardation syndrome complicated with bilateral sensorineural hearing impairment. PMID- 10453190 TI - Successful treatment of Epstein-Barr virus infection associated with myocarditis. PMID- 10453192 TI - Sotos syndrome associated with West syndrome and a visual disorder. PMID- 10453193 TI - Moya Moya syndrome in a child with hyperphosphatasia. PMID- 10453194 TI - Wilson's disease and Menkes disease. PMID- 10453195 TI - Mass screening for Wilson's disease: results and recommendations. AB - Wilson's disease is a treatable inherited disorder of copper metabolism. Established treatments include the use of oral chelating agents and the establishment of a minimum copper diet, although prognosis mainly depends on the extent of liver or nervous system damage present before treatment. Once irreversible damage has occurred, the effect of these treatments is diminished and the patient's quality of life compromised. Therefore, the establishment of a mass screening system able to detect Wilson's disease patients presymptomatically has been discussed. Recently, a monoclonal antibody specific to holoceruloplasmin has been developed. This antibody was used in a nationwide screening trial of 126,810 newborn infants, but no Wilson's disease patients were identified. However, three patients out of 24,165 were diagnosed with Wilson's disease using this specific antibody in a screening performed during the period from late infancy to elementary school. The age of 3 years is thought to be the best point for Wilson's disease mass screening. In this paper, a review of mass screening for Wilson's disease in Japan using a specific monoclonal antibody to holoceruloplasmin is presented. PMID- 10453196 TI - Molecular analysis and diagnosis in Japanese patients with Wilson's disease. AB - BACKGROUND: Wilson's disease is characterized by the toxic accumulation of copper in the liver, brain, cornea and other organs. It is caused by both impaired excretion via the bile and impaired incorporation of copper into ceruloplasmin in the liver. The Wilson's disease gene (ATP7B) has been cloned as a putative copper transporting P-type ATPase gene. We therefore analysed mutations of ATP7B in Japanese patients with Wilson's disease. METHODS: Twenty-three Japanese patients with Wilson's disease were investigated. In all patients, the ATP7B coding sequence, including exon-intron junctions, was analysed by restriction endonuclease digestion, mutation detected enhancement gel electrophoresis and/or direct sequencing analysis of amplified fragments. RESULTS: Thirteen mutations were identified, including seven missense mutations, four detections, one insertion and one exon skipping in the coding region. The most common mutations were 2874deletion(del)C in exon 13 and arginine (Arg)778 leucine (Leu) in exon 8. DISCUSSION: None of the observed mutations, except for 2302insertion(ins)C, have been previously detected in either European or North American patients. We conclude that the mutation spectrum of Wilson's disease may thus indicate a population-dependent pattern. Based on the population-dependent manner of the occurrence of ATP7B gene mutations, it may be possible to establish a molecular diagnosis system. A molecular diagnosis system is considered to be very effective for making a definitive diagnosis in very young patients and for also detecting carriers. PMID- 10453198 TI - Treatment and management of Wilson's disease. AB - Wilson's disease is an autosomal recessive disorder related to the copper metabolism. The clinical symptoms are due to copper deposition in various tissues, including liver, brain, kidney, cornea and others. The key strategy of treatment is to reduce the amount of copper in the liver and other tissues by administering both copper-chelating agents and a low copper diet. D-Penicillamine is considered to be the first choice as a copper-chelating agent. Patients require 15-25 mg/kg daily in the early stages of treatment and this drug should also be given more than 2 h before meals. Some undesirable or serious side effects, such as systemic lupus erythematosus (SLE) and nephrotic syndrome, do occur in 20-25% of all patients. In such cases, trienthylene tetramine (trientine) appears to be as effective as penicillamine. This drug is usually used when D-penicillamine has to be withdrawn. It is also sometimes administered to patients with neurological symptoms as a first-choice drug. It is given in doses of 40-50 mg/kg daily, in the same manner as for D-penicillamine. Zinc salt administration has also emerged as an interesting supportive therapy for both treatments. A dose of 5-7.5 mg/kg daily is given before meals. The copper content of the diet should be less than 1 mg/day in the early stages of treatment. Thereafter, it can be increased to 1.0-1.5 mg/day during well-controlled periods. Liver transplantation is now performed in many countries for patients with either the fulminant or chronic progressive types of Wilson's disease. PMID- 10453197 TI - The Long-Evans Cinnamon rat: an animal model for Wilson's disease. AB - The Long-Evans Cinnamon (LEC) rat is known to develop hepatitis and liver cancer spontaneously, phenomena attributed to abnormal copper metabolism. This mutant strain of rat shows some clinical features that are similar to those of Wilson's disease, including excessive copper in the liver, reduced excretion of copper into bile, a reduced level of serum copper and a remarkable decrease in serum ceruloplasmin activity. Molecular studies have revealed that the copper transporting P-type ATPase, atp7b, which is the rat gene homologous to human ATP7B, was found to be defective in the LEC rat. These observations have confirmed that the LEC rat is a rodent model for Wilson's disease. In addition, recent studies have suggested that the ATP7B protein is involved in the intracellular transport of hepatic copper. The absence or diminution of ATP7B function results in abnormal copper metabolism in the LEC rat and in patients with Wilson's disease. PMID- 10453200 TI - Molecular genetics and pathophysiology of Menkes disease. AB - The molecular genetics and pathophysiology of Menkes disease and an animal model for this disease are reviewed. The Menkes gene, located on chromosome X13.3, encodes a copper-transporting ATPase, as shown by the sequencing of a cDNA of 4500 bp. Mutations in the Menkes gene in patients with Menkes disease show great variety, including missense, nonsense, deletion and insertion mutations. Mutations in the Menkes gene have also been identified in patients with mild Menkes disease or occipital horn syndrome, showing that these diseases are allelic variants of Menkes disease. Mutations in the mottled gene, the murine homolog of the Menkes gene, have been demonstrated in mottled mutant mice that display biochemical and phenotypic abnormalities similar to those observed in patients with Menkes disease. In affected cells, copper significantly accumulates as metallothionein-bound copper in the cytosol and copper transport to the organelles, as well as copper efflux, is disturbed. As a result, cuproenzymes cannot receive the copper necessary for their normal function. Thus, the objective in treatment of Menkes disease and occipital horn syndrome is to deliver copper to the intracellular compartments where cuproenzymes are synthesized. PMID- 10453199 TI - Clinical manifestations and treatment of Menkes disease and its variants. AB - The clinical manifestations of classical Menkes disease, mild Menkes disease and occipital horn syndrome are reviewed. Menkes disease is a neurodegenerative disease with X-linked recessive inheritance. Orally administered copper accumulates in the intestine, resulting in the failure of copper absorption. The primary metabolic defect that causes copper accumulation in the intestine is present in almost all extrahepatic tissues. The blood, liver and brain are in a state of copper deficiency, which is due to defective copper absorption. The characteristic features, including neurological disturbances, arterial degeneration and hair abnormalities, can be explained by the decrease in cuproenzyme activities. DNA-based diagnosis is now possible. Mild Menkes disease and occipital horn syndrome, which show milder forms than Menkes disease, have been identified as genetic disorders resulting from mutations in the Menkes disease gene. Because the clinical spectrum of Menkes disease is wide, males with mental retardation and connective tissue abnormalities should be evaluated for biochemical evidence of defective copper transport. The treatment accepted currently is parenteral administration of copper. When treatment is started in patients with classical Menkes disease above the age of 2 months, it does not improve the neurological degeneration. When the treatment is initiated in newborn babies affected with this disease, the neurological degeneration can be prevented in some, but not all, cases. Moreover, early treatment cannot improve non neurological problems, such as connective tissue laxity. Therefore, alternative therapies for Menkes disease and occipital horn syndrome should be studied. PMID- 10453201 TI - Intracellular localization of the Menkes and Wilson's disease proteins and their role in intracellular copper transport. AB - Copper is a heavy metal ion essential for the activity of a variety of enzymes in the body. In excess, copper is a very toxic ion and therefore efficient regulation of its metabolism is required. This is dramatically illustrated by the genetic disorders X-linked Menkes disease and autosomal recessive Wilson's disease. In 1993, both the Menkes and Wilson's genes were isolated and it was found that these genes encode homologous cation copper transporting P-type ATPase proteins. The Menkes protein (ATP7A) is expressed in most tissues, except liver. In contrast, the Wilson's protein (ATP7B) is abundantly expressed in liver. Intracellular localization of those proteins was investigated. Both ATP7A and ATP7B are localized in the trans-Golgi network and post-Golgi vesicular compartment (PGVC) in the cell. This intracellular localization was altered by the copper content present in the cell. This result may support the hypothesis that ATP7A and ATP7B are involved in cellular copper transport and those proteins could be suitable models for elucidating intracellular copper metabolism. PMID- 10453202 TI - Evaluation of a rapid reagent strip test for the diagnosis of childhood meningitis. PMID- 10453203 TI - Plasmogastric passage of urea and ammonia in children with Helicobacter pylori infection. PMID- 10453204 TI - Diagnosis and therapeutic guidelines for hemolytic uremic syndrome associated with enterohemorrhagic Escherichia coli infection. Japanese Pediatric Nephrology Association. PMID- 10453205 TI - Catechol-O-methyltransferase inhibitors: new options for Parkinson's disease. PMID- 10453206 TI - [The statistics of dentistry: the state of the art, problems and the means for their solution]. PMID- 10453207 TI - [The characteristics of the oral health promotion of patients with hematological diseases]. AB - A total of 197 patients were subjected to planned sanitization of the oral cavity in St. Petersburg specialized dental hospital for short-term treatment. Critical values of blood parameters were determined for preventing the complications after planned sanitization. The probability of complications is lower if blood hemoglobin level is at least 100 g/liter, granulocyte count at least (1.5 2.0).10(9)/liter, platelet count at least 60.10(9)/liter, and erythrocyte content in patients with polycythaemia vera no more than (4.5-5.0).10(12)/liter. Preparation of patients to treatment carried out by a dentist together with a hematologist and total analgesia help eliminate stress reactions and effectively and timely correct the complications developing during treatment. This means that oral sanitization in patients with diseases of the blood system should be carried out at dental hospitals for short-term treatment. PMID- 10453208 TI - [Hemodynamic changes in the periodontium with the use of chewing gum]. AB - The effect of gum chewing (5 min after meals 3 times a day) on periodontal blood supply was studied in a group of 52 volunteers with different clinical status of the periodontium. Tonic strain in the local vessels changed (fluctuated) little (by 2-7%). Tonic strain was shifted on the inert side and in patients with periodontal diseases. Blood consumption increased by 33%. Regular gum chewing for 10 min resulted in compensatory vasoconstriction and hyperemia of the periodontium starting from the second week, irrespective of the initial clinical status of the periodontium. PMID- 10453209 TI - [Solcoseryl--dental adhesive paste in the treatment of stomatitis]. PMID- 10453210 TI - [The time characteristics of the exacerbation of chronic periodontitis and of odontogenic perimaxillary abscesses]. AB - Time course of some laboratory values and clinical manifestations of inflammatory process is studied in 52 patients with exacerbations of chronic periodontitis and 284 patients with odontogenic perimaxillary abscesses. Routine laboratory studies (clinical and biochemical analyses of the blood and urine) were carried out. Acute odontogenic inflammations are characterized by an intermittent course with periodicity of about a week. The end of the first week from the disease onset coincided with recovery in exacerbation of chronic periodontitis and with the beginning of stabilization of inflammatory process in abscess. Recovery after abscess was observed 2 weeks after disease onset. These regularities may be useful for specifying the pathogenesis, terms of check-ups, and duration of treatment. PMID- 10453212 TI - [A method for the accelerated preparation of cotton rolls for the isolation of the operating field from saliva]. PMID- 10453211 TI - [The lithotripsy of salivary stones]. AB - Salivary lithotripsy may be performed with lithotriptors used for crushing urinary concrements. In some cases the method helps escape removal of the salivary gland if the stone is localized inside the gland. Further modifications of the method are needed. PMID- 10453213 TI - [Experience with the treatment of keratocysts of the jaws]. AB - Thirteen patients aged 20-60 years (9 men and 4 women) with keratocysts of the jaws were observed. Clinical, x-ray, and microscopic picture of the disease is described. Keratocysts were removed and the resultant cavity scraped in 10 patients; in 3 the mandible was resected because of the process dissemination and vegetation of the periosteum and muscles. The diagnosis was confirmed morphologically. Enucleation with scraping is recommended in keratocysts; in extensive lesions with vegetation of tissues adjacent to the bone, the jaw is to be resected. PMID- 10453214 TI - [Assessing the results of the multimodal treatment of patients with combined jaw deformities]. AB - The results of treatment of patients with congenital combined deformations of the jaws are assessed after 6 months to 8 years. The results are analyzed for 96 patients with maxillary micro- or retrognathia and mandibular macro- or prognathia, maxillary macro- or prognathia, and mandibular micro- or retrognathia. Remote results were assessed by clinical, x-ray, and functional methods of examination. Stable esthetic and functional results were attained in all patients, which were observed during both immediate and remote periods of follow-up. No signs of deformation relapses were observed in any case. PMID- 10453215 TI - [Asymmetry of the facial skeleton in Negroid skulls]. AB - Measurements of 18 bilateral signs were carried out in 94 Negroid skulls. All parts of the facial skull are clearly asymmetrical, if regarded separately, while the summary sizes of the left and right halves of the facial skull are equal. In the upper part of the face the most asymmetrical parameters are the width of the bridge of the nose, width of the nose, and width of the palate, while the least asymmetrical are the height of the orbit and palate and length of the palate. In the mandibular part the greatest asymmetry was observed in the thickness and height of the mandibular corpus and height of the mandibular branches. The resultant combination of symmetry of the total sizes of both sides of the facial skull and asymmetry of individual parts is interpreted as an indication of the compensatory changeability of bilateral signs of the facial skull. The results are similar to data on the asymmetry of the facial skull of Mongoloid, Europeoid, and the intermediate skulls. PMID- 10453216 TI - [A method for the functional shaping of the internal surface of the base for a complete mandibular denture in the case of mobile mucosal folds]. AB - Functional lining of the internal surface of the basis of a complete mandibular denture is carried out for 21 patients aged 65-84 years with mobile folds of the mucosa on the denture bed. A more rapid accommodation (2-3 weeks) and improvement of the masticatory efficacy of dentures indicate that the method may be recommended for wide use. PMID- 10453217 TI - [Evaluation of the bioelectrical activity of the masticatory muscles in patients depending on the period since implantation]. AB - The results of electromyographic control used for evaluating the efficacy of orthodontic treatment are analyzed. Analysis of electromyographic parameters after orthodontic treatment of dentition defects with implants showed essential changes in the function of the masticatory muscles at different terms after implantation. Asymmetrical activity typical of unilateral chewing was observed, as well as high bioelectrical activity of the temporal muscles. PMID- 10453218 TI - [The demonstrative and information values of correlational methods in predicting the mesiodistal dimensions of the crowns of the canine teeth and premolars]. AB - Authors have analyzed the reliability of correlative methods with plaster-casts of 100 examines (50 boys and 50 girls) with permanent dentition, according to Moyers's and author's own methods, and methods of multiple regression analysis according to Bachmann and Traenkmann and collab., while anticipation of mesiodistal crown diameters of canines and premolar (C, P1, P2) in both jaws. The predictable amounts of crown width C, P1, P2 in the maxilla were larger than the measured on the model, according to the method: Moyers in 81% of cases, Legovic in 67% of cases, Bachmann in 35% and Traenkmann and collab. in 73% of cases. The predictable amounts of crown width C, P1, P2 in mandible were larger than the one measured on the model: Moyers in 81% of cases, Legovic in 80%, Bachmann in 45% and Traenkmann and collab. in 26% of cases, and smaller compared with the measured model: Moyers in 15% of cases, Legovic in 18%, Bachmann in 55% and Traenkmann and collab. in 74% of cases. PMID- 10453219 TI - [The extrusion of a retained upper central right incisor into the dentition]. AB - An extra tooth was the cause of retention of the upper right central incisor in patient S. After removal of the extra tooth and creation of conditions for pooling the 11th tooth by Angle's device, supplemented with a frame with hooks, the tooth was transposed into its proper position in the dentition. PMID- 10453220 TI - [The moral and legal aspects of the dentist's activities today]. PMID- 10453221 TI - [The use of gallium-based metal-containing filling materials as a replacement for mercury]. PMID- 10453222 TI - [Variants in the use of CAD/CAM systems in orthodontic dentistry]. PMID- 10453223 TI - [Advertising in dentistry: strategies, content, methods]. PMID- 10453224 TI - [The results and outlook for the use of mathematical methods and computer technology in dentistry]. AB - Development and introduction into wide practice of various mathematical methods (systemic, regression, and factor analyses, etc.) and modern computation devices (personal computers, local computer network, etc.) is shown. These devices extended the potentialities of differential diagnosis and pathogenetic (including laser) therapy of the major oral diseases (dental carries, pulpitis, periodontitis, periodontal diseases and buccal mucosa, odontogenic inflammations, tumors, etc.). PMID- 10453225 TI - [Trans-meatal hyperbaric aerosol therapy some forms of otitis media chronica]. AB - 80 patients with otitis media chronica were effectively treated with hyperbaric aerosol. The aerosol therapy was performed with the help of a special device the design of which was based on that of made in Russia ultrasonic inhalator IUP-OIM intended for aerosol therapy of the upper respiratory tracts. The inhalator was furnished with a mini-compressor for pumping aerosol in the middle ear with maximal permissible pressure of hyperbaric aerosol delivery 10 kPa. Such intratympanic pressure rules out the risk of ear barotrauma in the form of the cochlear window membrane rupture being at the same time sufficient for aerosol delivery into the nasopharynx through the mesotympanic defect of the tympanic membrane and the auditory tube, in secretory otitis--via previously made tympanostoma. The method provides also normobaric delivery of aerosol into the ear and therefore can be applied after tympanoplastic operations without the risk of the transplant dislocation. PMID- 10453226 TI - [Principles of uncomplicated epi- and epimesotympanitis treatment]. AB - The authors have treated more than 500 patients with uncomplicated epi- and epimesotympanitis. They described their approaches to surgical policy, plastic reconstruction after cleansing, performance of primary tympanoplasty without and with mastoidoplasty with formalinized cartilage, creation of the artificial antrum in the mastoid process. PMID- 10453227 TI - [Prolonged nasotracheal intubation in the treatment of respiratory tract scarring stenosis in children]. AB - Prolonged nasotracheal intubation lasting 24-153 days was applied for treatment of 12 children with forming and already established scarry stenosis of the respiratory tracts. The children had not previously undergone tracheotomy. The age of the children varied from 20 months to 7 years. Two thirds of them were admitted at the age under 3. Nine patients have developed stenosis after intubation for acute stenosing laryngotracheobronchitis. The treatment provided rather wide lumen of the respiratory tract in 10 patients. Tracheostoma was established only in 2 children. PMID- 10453228 TI - [Assessment of general and local immunity in children with affected lymphadenoid throat ring]. AB - 33 children with affected lymphadenoid throat ring and 15 healthy children were examined for local and general immunity status. Assessment of general immunity provided poor information. Measurements of secretion immunoglobulins levels with enzyme immunoassay was informative and demonstrated close similarity of the clinical and immunological indices. Concentrations of immunoglobulins in the lacunar secretion can serve a prognostic criterium. PMID- 10453229 TI - [Classification of auricle floor abnormalities in 1st and 2nd branchial arches syndromes]. AB - 1000 cases of the 1st and 2nd branchial arches syndrome and abnormalities of the floor of the auricle associated with this syndrome provided evidence for design of their classification. The classification distinguishes severe malformations of the floor of the auricle, malformations with tissue defect, deformations of the floor of the auricle itself. PMID- 10453230 TI - [Co-amoxiclav antibiotic therapy of acute otitis media, exacerbation of otitis media chronica and sinusitis in children]. AB - A comparative trial was performed of two regimens of antibiotic therapy in acute otitis, exacerbation of otitis media chronica and sinusitis in children. 100 children received combined treatment with amoxycilline and clavulanic acid (amoxiclav and autmentin). 50 control children were i.m. injected ampicilline. Oral co-amoxiclavs proved highly effective in ENT diseases in children and are superior to intramuscular ampicilline. PMID- 10453231 TI - [Orbital and intracranial complications of paranasal sinus inflammation]. AB - The examination of 2251 patients with paranasal sinusitis has detected various rhinosinusogenic orbital and intracranial complications in 75 of them (3.3% of overall number of sinusitis patients). The complications were caused by acute and chronic paranasal sinusitis in 49 and 26 patients, respectively. 55 patients had orbital, 20--different intracranial complications. The treatment in most cases consists of emergency operation on the paranasal sinuses, in intracranial complications the surgeon opens dura mater of the anterior cranial fossa and prescribes massive antibacterial, dehydration, desintoxication therapy. PMID- 10453232 TI - [Experimental choice of drugs for pharmacophysical impact on larynx]. AB - The authors validate the necessity of developing the method of laryngeal phonoelectrophoresis for treatment of acute and chronic laryngeal inflammation. Radioisotopes tracing shows high permeability of laryngeal tissue for drugs due to combined effects of phono- and electrophoresis. Maximal permeability of the dialysis membrane UB-20 was determined and optimal stereotypes of laryngeal phonoelectrophoresis with different drugs were proposed. PMID- 10453233 TI - [Efficiency of otoph and polydex ear drops in otitis treatment]. PMID- 10453234 TI - [Symptomatic therapy of pharyngeal pain]. PMID- 10453235 TI - [Anti-recurrence treatment of polypous rhinosinusitis]. AB - The authors propose a method of anti-recurrence treatment of polypous rhinosinusitides which implies drop introduction and alternating of antihistamine and antikinin drugs. The results of the treatment show that the number of recurrences reduces, remission duration increases. PMID- 10453237 TI - [Pilot experience with radio-knife "Surgitron"]. AB - A new radiosurgical knife Surgitron is described in terms of its application in ENT operations. Surgical procedures, indications, operative regimes are detailed. Advantages and disadvantages of the device design are outlined. The device is functionally and cosmetically effective. Thus, it holds promise in surgery. PMID- 10453236 TI - Noval radiosurgical technique of cuts and hemostasis in otorhinolaryngology. AB - A novel radiosurgical method of cutting tissues and coagulation of ENT operations is presented. Indications, technique, device operation regimes are provided. Radiosurgery has evident advantages: saving cut with minimal damage to the adjacent tissues, good coagulating effect warranting "dry field" for the surgeon, absence of necrosis of the wound walls, fast healing, simple operative procedure. Radiosurgery holds great promise for otorhinolaryngological practice. PMID- 10453239 TI - [Combination of bilateral otitis media chronica purulence and hemorrhagic fever with renal syndrome]. PMID- 10453238 TI - [Local frontobasal leptomeningitis in patient with pyocele of frontal sinuses]. PMID- 10453240 TI - [Acute tonsillogenic thrombosis of right jugular vein]. PMID- 10453241 TI - [Rare location of rhinogenic cerebral abscess]. PMID- 10453242 TI - [A case of aspirin triad]. PMID- 10453243 TI - [Some aspects of scleroma problem]. PMID- 10453244 TI - [Magic and religion in medicine and otorhinolaryngology]. PMID- 10453245 TI - Continuously recorded suckling behaviour and its effect on lactational amenorrhoea. AB - The hypothesis that the month-specific rate of return to ovarian cyclicity after childbirth is causally related to suckling pattern was tested for a population of New Mexican women recruited within the service area of New Mexico Highlands University and for a nationwide USA subpopulation of women recruited through membership of the Couple to Couple League (CCL). Survival analysis for time dependent covariates was used, and significant predictors of the first postpartum menses were found. Important differences were detected in the suckling pattern for the two groups and a 5:2 differential was found in their respective rates of menstrual cycle recovery. Although the two groups were comparable perinatally, daily and time-windowed breast-feeding performance fell off at twice the rate for the New Mexico population when contrasted with the CCL sample. For both populations, the introduction of solid feeds was a strong and significant predictor of returning menstrual cyclicity, independent of suckling pattern. PMID- 10453246 TI - Area variations in use of modern contraception in rural Bangladesh: a multilevel analysis. AB - This study in Bangladesh found that inter-cluster variation in the use of modern reversible methods of contraception was significantly attributable to the educational levels of the female family planning workers working in the clusters. Women belonging to clusters served by educated workers had a higher probability of being contraceptive users than those whose workers had only completed primary education. At the household level, important determinants of use were socioeconomic status and religion. At the individual level, the woman being the wife of the household head and having some education were positively related to her being a user. The model also found that inter-household variation was significantly greater than inter-cluster variation. Finally, the study concludes that after controlling for various covariates at all three levels, the clusters do not have significantly different levels of use of modern reversible methods of contraception. There are, however, some special areas where contraceptive use is dramatically low, and these contribute significantly to the observed inter cluster variation. PMID- 10453247 TI - Determinants of contraceptive discontinuation in six developing countries. AB - This analysis investigates the determinants of contraceptive discontinuation in six developing countries, using data from Phase I surveys of the DHS programme. Cumulative probabilities of discontinuation at 24 months for reasons other than the desire for another child were examined. By this time, typically about 40% of couples have stopped use and most are subsequently at risk of an unwanted conception. Discontinuation of IUD use was found to be less common than for other methods, partly perhaps because cessation of use requires a deliberate decision to have the device removed. The most important results are negative ones. Neither the schooling of couples nor their type of residence exerted appreciable influence on discontinuation. The policy and programme implications are discussed. Prior use of a method, fertility preferences and the related demographic factors of age and family size emerged as pervasive predictors of discontinuation. PMID- 10453250 TI - Child undernutrition in war-torn society: the Ethiopian experience. AB - Using data from a cross-sectional nutrition survey conducted in rural Ethiopia between March and April 1992, roughly a year after the end of one of the longest civil wars in modern human history, this study attempts to document the magnitude and correlates of childhood undernutrition in Ethiopia. Findings from the study reveal that, at the time of the survey, 59% of children in the country were exposed to long-term or chronic undernutrition (stunted); about 4% were suffering from acute problems (wasted); and about the same proportion were both stunted and wasted. Less than a third of the country's children had normal growth. The logistic-binomial regression results demonstrated the existence of significant clustering of risks of undernutrition within areas of residence and notable differentials by age of child, duration of breast-feeding, age at introduction of supplementary foods and number of under-five siblings. Significant variations were also noted by age of household head, per capita land holding, religious affiliation and prevalence of endemic diseases in an area. PMID- 10453249 TI - Psychosocial stress and the menstrual cycle. AB - The relationship between mood states, urinary stress hormone output (adrenaline, noradrenaline and cortisol) and adequacy of the menstrual cycle was examined in 120 recorded non-conception cycles from 34 women. It was hypothesized that women with higher stress levels would be more likely to experience abnormal cycles and that within women higher stress levels would positively relate to follicular phase length and inversely relate to luteal phase length. There was a non significant trend for women to report higher stress levels during oligomenorrhoeic and unclear cycles compared with normal cycles. Analysis of covariance indicated that there was no consistent relationship between the measures of stress used here and follicular or luteal phase length within women. There was also no consistent pattern of relationship between reported mood states and stress hormone excretion within women. Further research is warranted to understand the role of stress and subtle menstrual cycle abnormalities in female fertility. PMID- 10453251 TI - Social contrasts in the incidence of obesity among adult large-city dwellers in Poland in 1986 and 1996. AB - The incidence of obesity, defined as the fraction of persons with BMIs exceeding 30-0, was examined in two birth cohorts of 40-50-year-old occupationally active inhabitants of the city of Wroclaw, Poland. In both cohorts and both genders obesity increased monotonically with decreasing position on a three-level educational scale. During the 1986-1996 decade obesity increased dramatically among males with trade school education only; concomitant shifts were much smaller or absent in the college-educated groups. The contrast in obesity between the opposite ends of the educational scale has widened markedly in both genders; and the between-gender gap has declined somewhat. PMID- 10453248 TI - Adolescent transition to coitus and premarital childbearing in Sudan: a biosocial context. AB - This paper examines the biosocial basis of premarital sexual and reproductive behaviour among women in Sudan. It applies Udry's biosocial perspective, which attempts to reconcile the biological and sociological models of premarital sexual and reproductive behaviour. World Fertility Survey (WFS) data were used to study premarital first motherhood. Early puberty was found to be paramount in determining childbearing in a separate biological model, but also in a biosocial model constructed to take account of social controls. This finding suggests that social controls do not influence the biological predisposition to premarital sexual behaviour. However, given the limitations of the WFS data, conclusive evidence must await a more appropriately designed study of reproductive behaviour in Sudan. PMID- 10453252 TI - Cancer, genomics, and the National Cancer Institute. PMID- 10453253 TI - Presentation of the Kober Medal for 1999 to Jean D. Wilson physician-scientist exemplar. PMID- 10453255 TI - Medical considerations for wilderness and adventure travelers. AB - Wilderness and adventure travel has attracted a growing number of participants. Many adventure travelers are inadequately prepared for their trip and are naive about the associated risks. Physicians are frequently asked to provide medical clearance and to complete medical release forms to travelers. To provide better advice, the physician should be familiar with the rigors and difficulty of these activities and must consider the potential environment hazards and remoteness of the intended area of travel. This article provides information on trip rating scales, environmental hazards, altitude illness, hypothermia, heat illness, and adventure travel medical kits. PMID- 10453254 TI - Emerging infectious diseases and risk to the traveler. AB - This article examines the relationship between travel and emerging infections. The authors begin with an overview of disease emergence and follow with a brief infection-by-infection examination of selected emerging pathogens of particular relevance to travelers and the medical care providers who counsel them. Emphasis is given to those agents that clearly have emerged as significant new or increased risk to travelers; or are of sufficiently new interest, even in the face of inadequate data in travelers, to be of potential immediate concern. The authors also discuss several novel pathogens, such as Ebola virus, that are clearly of insignificant or minimal risk to travelers, but are the subject of frequent questions from patients requesting pre-travel advice from medical providers. PMID- 10453256 TI - Travel immunizations. AB - An updated approach to selecting and prioritizing immunizations for the international traveler is presented. This article addresses vaccines against yellow fever, typhoid fever, cholera, meningococcal meningitis, rabies, tetanus, diphtheria, measles, mumps, rubella, polio, varicella, and influenza. Vaccine preparations, dosing regimens, efficacy, adverse effects, indications, and contraindications are discussed in the context of pre-travel preparation. PMID- 10453257 TI - Prevention of malaria in travelers. AB - Malaria occurs throughout the tropics and represents a serious health threat to people exposed to risk of infection. Health care providers may be the only link between information for their traveling patients and that threat. The likelihood of infection can be reduced drastically by deliberate measures that minimize exposure to biting mosquitoes. However, personal protective measures alone rarely suffice where exposure to anopheline mosquitoes and infected human beings is appreciable. PMID- 10453258 TI - Prevention and empiric treatment of traveler's diarrhea. AB - Diarrheal illness has plagued travelers for centuries, and to this day it remains the most common medical problem affecting travelers from industrialized nations to areas of the world where substandard hygiene conditions and sanitation prevail. This article discusses the cause of diarrheal illness in travelers, as well as epidemiology, prevention, treatment, and a general approach to self treatment. PMID- 10453259 TI - Global risk of sexually transmitted diseases. AB - Travelers returning from international destinations may have particular concerns regarding their risk of having acquired sexually transmitted diseases (STDs). The authors review diagnosis, treatment, and special considerations regarding viral, bacterial and protozoan STDs in the international traveler. Diseases that mimic STDs are presented. The authors also discuss the importance of preventative measures and pre-travel counseling. PMID- 10453260 TI - Evaluation of fever in the returned traveler. AB - The differential diagnosis of a febrile illness in the returned traveler is extensive. The most commonly encountered tropical infections are malaria, dysentery, hepatitis, and dengue fever; a substantial number of febrile illnesses are never diagnosed. Malaria is by far the most important infection to consider in the returned traveler who presents with fever. As international travel continues to increase in popularity, the ongoing need for clinicians to broaden their knowledge of travel-related diseases is evident. The ability to recognize and manage tropical diseases in travelers is essential because the morbidity and mortality of these infections are often preventable with prompt therapy. When expertise in this area is lacking, febrile returned travelers should be referred to a tropical disease unit or an infectious disease consultant for urgent assessment. PMID- 10453261 TI - Eosinophilia in the returning traveler. AB - The most common cause of eosinophilia in the traveler returning from the developing world is from infection with a helminth parasite. Protozoal, bacterial, and fungal infections are uncommon causes of eosinophilia. Allergic disorders and drug reactions must also be considered, as well as numerous uncommon miscellaneous disorders. This article reviews the diagnostic features of the more common causes of eosinophilia in the returned traveler and suggested drug treatment. PMID- 10453262 TI - Chronic diarrhea in the returned traveler. AB - The persistence of gastrointestinal symptoms after travel to a developing country is one of the most common and troublesome post-travel illnesses. Few data exist to document the extent of this problem, but anecdotal examples abound among travel medicine practitioners, internists, and gastroenterologists. This article presents an approach to the patient with persistent gastrointestinal symptoms after travel. PMID- 10453263 TI - Intestinal parasites in returned travelers. AB - Parasites in the returned traveler can present in any organ system and at time periods quite distant from the activity or geographic exposure that created the risk for infection. All clinicians need to be aware that the most efficient approach to these unusual diseases starts with taking a travel history. The exciting challenge to the diagnostician and the laboratorian is to be able to respond to these clinical demands. PMID- 10453264 TI - Skin lesions in returned travelers. AB - Skin problems are commonly encountered during and after travel. Because of increasing travel and tourism to remote and previously unvisited areas, it is likely that these and other dermatologic conditions will continue to be frequent problems observed in travelers. For the nondermatologist, recognition of many of the commonly encountered skin problems can be facilitated by following the lesion oriented approach outlined in this article. PMID- 10453267 TI - 4th International Stem Cell Workshop: High dose therapy and transplantation of haemopoietic stem cells and Round Table discussion of experts in stem cell transplantation: Indications and novel approaches in allogeneic and autologous blood cell transplantation. Bad Saarow/Berlin, Germany, April 29-May 1, 1999. Abstracts. PMID- 10453266 TI - [XXXII National Congress of the Spanish Society of Pneumology and Thoracic Surgery. Madrid, Spain, 15-18 May 1999. Abstracts]. PMID- 10453265 TI - Refugee and immigrant health care. AB - Health care providers involved in caring for immigrants and refugees must be aware of overseas and domestic screening protocol recommendations, common disease entities seen in these patient populations as well as those diseases with long latency periods that may appear years to decades after U.S. arrival. This article examines these and other topics. PMID- 10453268 TI - ISIR and JSAIR'99. 7th International Symposium on Interventional Radiology and New Vascular Imaging. 28th Annual meeting of the Japanese Society of Angiography and Interventional Radiology. Osaka, Japan, May 16-19, 1999. Abstracts. PMID- 10453269 TI - 4th Meeting of the European Society of Neurosonology and Cerebral Hemodynamics. Venice, Italy, April 10-13, 1999. Abstracts. PMID- 10453270 TI - Gastrointestinal pathology. 83rd Meeting of the German Society of Pathology. Jena, Germany, 26-29 May 1999. Abstracts. PMID- 10453271 TI - Annual scientific meeting of the Thoracic Society of Australia and New Zealand. Canberra, Australia, 26 February-3 March 1999. Abstracts. PMID- 10453272 TI - [Ontology and good practices in epidemiology. Guidelines. ADELF, ADEREST, AEEMA, EPITER]. PMID- 10453273 TI - Targeting angiogenic pathways involving tumor-stromal interaction to treat advanced human prostate cancer. AB - Interfering with and preventing tumor angiogenesis is an attractive therapeutic approach for treating cancer metastases. This commentary presents treatment strategies that may enhance the effectiveness of anti-angiogenic therapy by selectively targeting newly sprouting and immature vessels, inhibiting the production of angiogenic factors, and disrupting extracellular matrices. We propose several clinical paradigms, including hormonal ablation, intermittent androgen suppression, chemotherapy, and radiation therapy, that 'injure' nascent vasculature and interrupt the cancer cell-stromal relationship, thereby potentiating the efficacy of experimental anti-angiogenic agents. These stromal epithelial interactions play an important role in the development, proliferation and dissemination of prostate cancer, as well as guiding the processes of tumor neovascularization. Successful utilization and targeting of tumor angiogenesis requires an increased understanding of tumor cell-stromal cell-endothelial cell relationships, most notably the intricate intracellular signalling cascades mediated by growth factors and the extracellular matrix. PMID- 10453274 TI - Peptide growth factors and prostate cancer: new models, new opportunities. AB - Prostate cancer is the leading form of newly diagnosed cancer cases in men in the United States. However, the molecular mechanisms contributing to the initiation, progression and ultimate development of metastatic and androgen independent disease are poorly understood. This is due in part to the difficulty in obtaining clinical samples representing early disease and the lack of animal models that recapitulate the full spectrum of the clinical disease. To this end we have developed and characterized the Transgenic Adenocarcinoma of Mouse Prostate (TRAMP) animal model that expresses the oncogene SV40 T antigen specifically in the epithelium of the prostate. TRAMP develops spontaneous autochthonous prostate cancer compelte with distant site metastasis and can progress to androgen independent disease. Changes in the fibroblast growth factor (FGF) axis and the insulin-like growth factor (IGF) axis have been examined during prostate cancer progression utilizing the TRAMP model and these data generally support observations reproted in the clinical disease. Moreover, we report novel changes in the FGF axis and IGF axis utilizing TRAMP. Thus, TRAMP can be used as a potent tool in understanding the mechanism of prostate cancer initiation and progression. PMID- 10453275 TI - Diet, androgens, oxidative stress and prostate cancer susceptibility. AB - Prostate cancer is the most common human malignancy and the second leading cause of cancer deaths among men in Western nations. Descriptive epidemiologic data suggest that androgens and/or environmental exposures, such as diet (in particular, dietary fat), play an important role in prostatic carcinogenesis. One plausible link between diet and prostate cancer is oxidative stress. This process refers to the generation of reactive oxygen species, which can then trigger a host of pro-carcinogenic processes. Recent studies also indicate that androgens increase oxidative stress within human prostate cancer cell lines. Recent data from our institution indicate that oxidative stress is higher within the benign epithelium of prostate cancer patients than men without the disease. This confirms our hypothesis and suggests that antioxidants such as lycopene, vitamin E, and selenium may play an important role in preventing disease progression. Large-scale clinical trials with some of these agents are currently in the design phase. PMID- 10453276 TI - Mechanisms, hypotheses and questions regarding prostate cancer micrometastases to bone. AB - The morbidity and mortality associated with prostate cancer can almost universally be attributed to the consequences of metastases to the bone. While clinically there have been descriptive reports of these lesions and their detection by bone scan, there is an embrrassing paucity of reports as to the mechanisms of prostate cancer cell trafficking to the bone, adaptation to the bone environment, pertubation of the normal bone reformation process and the events leading to cachexia and death. In recent years, there have been numerous in vitro studies suggesting that PSA and hK2 may play a significant biological role in these events. Also, recent data generated form reverse transcription polymerase chain reaction assays reveal that metastasis to the bone may be an early event which further underscores the need to better understand this complex and critically important process. This commentary highlights several general concepts and a few specific issues related to CaP bone metastasis with the intent of revealing numerous opportunities for further investigation and inquiry. PMID- 10453277 TI - Role of p27 in prostate carcinogenesis. AB - The cyclin dependent kinase inhibitor p27 binds to and inhibits preferentially S phase kinases thereby halting cell cycle progression. Loss of p27 expression has been shown to be associated with aggressive behavior in a variety of human epithelial tumors including prostate cancer. In this review, the role of p27 in cell cycle progression as well as its regulation by the ubiquitin-proteasome pathway are discussed. The experimental evidence pointing to the role of p27 as a tumor suppressor gene is outlined. The data generated to date on the prognostic significance of loss of p27 protein expression in human prostate cancers are summarized. Finally, the implications of the changes in p27 expression which occur as a result of androgen ablation in normal and neoplastic prostate are discussed. PMID- 10453278 TI - Molecular regulation of cell death and therapeutic strategies for cell death induction in prostate carcinoma. AB - Many of the common molecular alterations associated with prostate cancer progression involve genes known to regulate cell death susceptibility. The significance of these molecular events is discussed in the context of developing and implementing new strategies designed to restore cell death susceptibility in prostate cancer cells and overcome therapeutic resistance. PMID- 10453279 TI - Vitamin D and prostate cancer: biologic interactions and clinical potentials. AB - While prostatic cancer is usually thought of as an androgen dependent disease, it has recently become clear that numerous other hormones including 1 alpha,25 dihydroxyvitamin D3 (1,25D) can regulate its growth and differentiation. This commentary reviews the epidemiological evidence supporting a role for 1,25D deficiency in the genesis of prostatic cancer, the effects that this hormone has on benign and malignant prostatic epithelial cells in culture, the mechanisms through which it produces these effects and the potential clinical utility of 1,25D and its non-hypercalcemic analogs in the prevention and treatment of this increasingly common disease. While a wide body of information exists regarding the actions of vitamin D in other organ sites, this review addresses only prostatic studies. The substantial nature of the current information suggests that we have arrived at a point where we can no longer think of prostate cancer solely in terms of its response to androgens. PMID- 10453280 TI - Model systems of prostate cancer: uses and limitations. AB - A valid experimental model system reflects the system under study and is reproducible. Model systems of prostate cancer that accurately reflect the different disease stages are necessary to ensure a proper experimental design aimed at increasing our understanding of the biology of the disease and such models are essential tools to accelerate development of new therapies for prostate cancer. Until recently, a limited number of experimental systems were available and more suitable models derived from human specimens have only recently been developed and become available for use. In addition, transgenic techniques have also permitted the development of unique mouse models. The difficulty in establishing model systems may reflect the complex requirements necessary for cancer progression and should lead us to interpret results from model systems with caution. It is unlikely that a single model system that faithfully reflects the whole process of cancer development and progression will be developed. However, thoughtful use of the available model systems will permit the study of a significant portion of prostate cancer progression. In this review we summarize the properties of the prostate cancer model systems in use and defined their utility and limitations. This review will guide the investigator seeking models with which to test specific hypotheses pertaining to prostate cancer. PMID- 10453281 TI - Hormone resistance in prostate cancer. PMID- 10453282 TI - Insulin-like growth factors and prostate cancer. AB - Converging data from epidemiological and biological research implicate insulin like growth factor (IGF) physiology in the regulation of prostate epithelial cell proliferation and in the pathophysiology of prostate cancer. This review (1) outlines elements of IGF physiology, (2) reviews recent evidence that circulating IGF-I level is related to risk of prostate cancer, (3) provides a hypothesis concerning the biological basis for the relationship between IGF-I level and risk of prostate cancer, (4) discusses IGF-I physiology in the context of neoplastic progression of prostate cancer, and (5) discusses clinical implications of these lines of research with respect to prevention and treatment. PMID- 10453283 TI - Stem cell genes in androgen-independent prostate cancer. AB - Despite recent advances in the detection and treatment of early stage prostate cancer, there remains little effective therapy for patients with locally advanced and/or metastatic disease. Although the majority of patients with advanced disease respond initially to androgen ablation therapy, most go on to develop androgen-independent tumors that are inevitably fatal. Therefore, understanding the mechanisms by which a hormone-sensitive tumor escapes hormonal control is critical to the development of effective therapeutic modalities. The study of the differentiation pathways of normal and abnormal prostate growth has led to the development of a stem cell model for prostate cancer [1-3]. Recent work discussed in this commentary suggests that prostate tumors resist apoptosis and proliferate by adopting features of normal prostatic stem/progenitor cells. Basal cells, the putative stem/progenitor cells of the prostate, possess the phenotype of androgen independence as do most advanced prostate cancers. Therefore, the study of basal cells may prove critical to understanding prostate carcinogenesis and to the development of novel strategies for preventing and managing prostate cancer. PMID- 10453284 TI - Epigenetic mechanisms for progression of prostate cancer. AB - Epigenetic mechanisms may be the main driving force for critical changes in gene expression that are responsible for progression of prostate cancers. The three most extensively characterized mechanisms for epigenetic gene-regulation are (i) changing patterns of DNA methylation, (ii) histone acetylations/deacetylations, and (iii) alterations in regulatory feedback loops for growth factors. Several studies have indicated that DNA hypermethylation is an important mechanism in prostate cancer for inactivation of key regulatory genes such as E-cadherin, pi class glutathione S-transferase, the tumor suppressors CDKN2 and PTEN, and IGF II. Similarly, histone acetylations and deacetylations are frequently associated respectively with transcriptional activation (e.g. IGFBP-2 and p21) and repression (e.g. Mad:Max dimers) of genes linked to prostate cancer progression. Recently, histone acetyltransferase and deacetylase activities have been shown to be intrinsic with transcriptional coregulator proteins that bind to steroid receptors (e.g. SRC-1 and PCAF). Changes in regulatory feedback loops for growth factors with prostate cancer progression tend toward shifts from paracrine to autocrine control where the receptor and ligand are produced by the same cell. While there are several examples of this progression pattern in prostate tumors such as with IGF, FGF, TGF-alpha and their respective receptors, the precise mechanism (i.e. epigenetic or mutational) is less certain. In the context of treatment options, the contribution of mutational versus epigenetic events to prostate cancer progression is an important consideration. Irreversible genetic changes are likely to be less amenable to therapeutic control than are epigenetic ones. PMID- 10453285 TI - What might a stromal response mean to prostate cancer progression? AB - Many similarities exist between the stroma at sites of wound repair and reactive stroma in cancer. Common features include an elevated stromal cell proliferation, altered expression of matrix components, elevated expression of TGF beta-1, neovascularization, and expression of several common stromal markers. In addition, proliferative stromal cells at these sites generally express myodifferentiation markers. A comparison between the many common features and the biologically active molecules observed in reactive stroma in carcinoma and reactive stroma in wound repair is discussed in this review. An extended analysis of the literature suggests a functional link between mechanisms in wound repair response and the stromal reaction in many cancers including prostate cancer. We propose in this review, that the fundamental mechanisms of stroma in providing a rapid response to altered homeostasis in wounding, also provides for a tumor regulating stromal microenvironment in cancer. The functional consequences of this stromal response to carcinoma progression and how the stromal response might be used in extended diagnosis and in therapeutic approaches are discussed. PMID- 10453286 TI - Mechanistic concepts in androgen-dependence of prostate cancer. AB - Androgen blockade is the mainstay of therapy in the clinical management of advanced prostate cancer. Recent progress on two fronts--the development of newer xenograft and transgenic models and a greater understanding of nuclear receptor signaling--has provided new insight into mechanisms of androgen-dependence in prostate cancer. This review centers on the concept that perturbations in androgen receptor signaling are likely to occur early in prostate cancer and play a critical role in progression to end stage hormone-refractory disease. PMID- 10453287 TI - Prognostic markers in localized prostate cancer: from microscopes to molecules. AB - Management of patients diagnosed with localized prostate cancer is complicated by the diverse natural history of the disease and variable response to treatment. Prognostic criteria currently in use cannot fully predict tumor behavior and thus limit the ability to recommend treatment regimens with the assurance that they are the best course of action for each individual patient. The search for better prognostic markers is now focussed on the molecular mechanisms which underlay tumor behavior, such as altered cell cycle progression, apoptosis, neuroendocrine differentiation, and angiogenesis. As the number of potential molecular markers increases, it is becoming evident that no single marker will provide the prognostic information necessary to make a significant improvement in patient care. In addition, it seems likely that traditional methods of assessing the prognostic value of this multitude of new markers will prove inadequate. In this review, we briefly examine the current state of prognostication in localized prostate cancer and some of the promising new molecular markers. Next, we examine how new technologies may allow the multiplex analysis of vast numbers of markers and how computational methods such as artificial neural networks will provide meaningful interpretation of the data. In the near future, such an integrated approach may provide a comprehensive prognostic tool for localized prostate cancer. PMID- 10453288 TI - Metastasis-related genes in prostate cancer: the role of caveolin-1. AB - Prostate cancer continues to be a significant cause of death in U.S. men despite screening of asymptomatic men and extensive treatment with potentially curative therapies, predominantly radical prostatectomy and irradiation therapy. The reason for the persistent mortality resides in part in the presence of occult metastases at the time of treatment. Currently there are no curative therapies for metastatic prostate cancer. To better understand the metastatic phenotype in prostate cancer, we developed a strategy to compare and isolate mRNAs that are expressed differentially in cell lines derived from primary versus metastatic mouse prostate cancer using differential display-PCR. This strategy has proven to be successful and multiple gene sequences associated with metastasis in this model are being investigated. One of the genes isolated by this method was caveolin-1. Caveolin-1 was found to be overexpressed not only in metastatic mouse prostate cancer, but also human metastatic disease. Recent studies have indicated that suppression of caveolin expression induces androgen sensitivity in high caveolin, androgen-insensitive mouse prostate cancer cells derived from metastases. Overexpression of caveolin leads to androgen insensitivity in low caveolin, androgen-sensitive mouse prostate cancer cells. Caveolin-1, therefore, is a metastasis-related gene and a candidate gene for hormone-resistant prostate cancer in man. PMID- 10453290 TI - The role of cell motility in prostate cancer. AB - Cell motility is a critical determinant of prostate cancer metastasis. The current review discusses the role for cell motility in metastatic dissemination, the evidence that prostate cancer metastasis is dependent on increased cell motility and describes the molecules whose expression has been shown to correlate with the increased motility that accompanies prostate cancer progression. These include receptors for growth factors and cytokines that regulate cell motility as well as intracellular proteins that interact with actin or that regulate signal transduction associated with cell motility. Motility related modulators include both positive regulators of cell movement that are upregulated during tumor progression and suppressors of cell movement that are down-regulated during progression. Because altered expression of such genes may determine the metastatic potential of any particular prostate tumor, we conclude that the appearance or disappearance of motility-related molecules could be used to aid in the diagnosis and prognosis of human prostate cancer. PMID- 10453289 TI - Growth factor network disruption in prostate cancer progression. AB - Prostate cancer progression is often accompanied by alterations in expression of one or more growth factor receptors and/or their ligands. This commentary examines the hypothesis that the disruption of the normal growth factor network itself is a critical event in prostate cancer progression. If such transition points are proven, there are significant ramifications. PMID- 10453291 TI - The evolution of stress echocardiography. AB - The feasibility of using echocardiography to identify stress induced wall motion abnormalities was first demonstrated with M-mode recordings. The practical use of such a test had to await the development of 2-dimensional echocardiography whereby more wall segments could be analyzed. From the early days of 2 dimensional echocardiography there have been a succession of technological and clinical advances which have made stress echocardiography a very clinically useful tool in the management of patients with known or suspected coronary artery disease. These developments included the realization that stress-induced wall motion abnormalities produce stunned myocardium permitting immediate posttreadmill echoes to be clinically useful, the use of pharmacologic stress, the introduction of digital recording techniques so that rest and stress images could be viewed side-by-side, and more recently the advent of new imaging technologies, such as harmonic imaging of tissue to provide higher quality of stress echocardiograms. PMID- 10453292 TI - Exercise echocardiography. Principles, methods, and clinical use. AB - Stress echocardiography is composed of a family of examinations in which various forms of cardiovascular stress are combined with echocardiographic imaging to assist in the diagnosis of coronary artery disease. Exercise cardiography has evolved over the past 20 years into a routinely available clinical tool employed in both university and community hospital settings. This article discusses advantages and disadvantages of using exercise echocardiography. PMID- 10453293 TI - Pharmacologic stress echocardiography. Dobutamine and arbutamine stress testing. AB - Pharmacologic stress testing is an important noninvasive method for evaluating patients with known or suspected coronary artery disease who are unable to adequately exercise. Pharmacologic stress echocardiography using dobutamine has been developed over the last 10 to 15 years as an alternative to vasodilator stress testing using nuclear perfusion imaging. As experience has grown, digital subtraction echocardiogram has been shown to be a safe, convenient, and reliable method for stress testing in a variety of patient populations. Digital subtraction echocardiogram has comparable sensitivity, specificity, and accuracy when compared to other stress testing methods which employ cardiac imaging and is superior to the exercise echocardiogram. It has certain advantages over nuclear perfusion imaging in terms of cost and convenience. The recent addition of arbutamine echocardiography (which has been shown to be comparable to digital subtraction echocardiogram) provides another alternative method for pharmacologic stress testing. Continued improvement in echocardiographic image quality and the development of new technologies such as tissue harmonic imaging and contrast echocardiography will hopefully improve the echocardiographic evaluation of wall motion therefore increasing the diagnostic accuracy of echocardiographic stress testing. PMID- 10453295 TI - Transesophageal echocardiography with stress for the evaluation of patients with coronary artery disease. AB - Echocardiography permits a comprehensive assessment of resting regional and global left ventricular function, the presence and extent of inducible myocardial ischemia, and the identification of myocardial viability. Accordingly, stress echocardiography has become a valuable tool for the evaluation of patient with known or suspected coronary artery disease. In some patients however, a suboptimal transthoracic echocardiogram may limit the performance of interpretation of the test. Transesophageal echocardiography in combination with stress has been recently used for the evaluation of patients with coronary artery disease. This technique is semi-invasive, more time-consuming, and requires a greater degree of expertise on the part of the personnel assisting with the test. In general, complications and side-effects are self-limited and rarely affect the diagnostic accuracy of the test. Based on its ability to provide high quality images, transesophageal stress echocardiography should be considered in patients who have suboptimal transthoracic ultrasound window for the quantitative assessment of myocardial wall-thickening in clinical investigations of ischemic heart disease. PMID- 10453294 TI - Dipyridamole stress echocardiography. AB - Dipyridamole stress is the forerunner and prototype of pharmacological stress echo tests in the diagnosis of coronary artery disease. The safety of this test has been conclusively demonstrated as a result of extensive experience in large scale multicenter projects. The diagnostic accuracy of dipyridamole stress echo is comparable to dobutamine and largely a function of the employed dose. Higher dosages (up to 0.84 mg/kg) are being required to achieve good sensitivity. The prognostic value has been shown to be independent and additive to clinical, exercise echocardiogram, and angiographic data. The test positive response should be titrated on the basis of severity, extent, and timing of induced dyssynergy with low positivity being associated to more anatomically and functionally severe forms of disease. Multicenter, randomized, prospective, international studies on cost-effectiveness directly comparing a noninvasive strategy centered on stress echo versus an invasive strategy centered on coronary angiography are currently ongoing. PMID- 10453296 TI - Use of stress echocardiography for risk assessment of patients after myocardial infarction. AB - The main predictors of outcome after infarction (exercise capacity, ejection fraction, and extent of jeopardized myocardium) can all be identified using stress echocardiography. This review addresses the place of stress echocardiography in postinfarct risk evaluation, relative to clinical evaluation, and other technologies. The test is accurate for identification of multivessel disease and for predicting outcomes, is versatile, and can be used early after infarction. PMID- 10453297 TI - Assessment of myocardial viability with stress echocardiography. AB - Over the past two decades, there has been an increased realization that systolic myocardial dysfunction, outside of the setting of acute ischemia, does not necessarily imply irreversible myocardial injury. Echocardiographic techniques, particularly dobutamine stress echocardiography, have emerged as important diagnostic modalities that can identify residual viable myocardium in patients following acute myocardial infarction and in those with suspected myocardial hibernation. Dobutamine echocardiography can also help risk stratify patients with coronary artery disease and depressed ventricular function and identify patients who would benefit best from revascularization procedures. PMID- 10453298 TI - Stress echocardiography in valvular heart disease. AB - Stress echocardiography has been widely accepted as an important diagnostic and prognostic tool in the assessment of known or suspected coronary artery disease. Its use in valvular heart disease, to date, has been more limited, but is continuing to grow as the technology and the understanding of valvular disorders progress. In this article, we will review the current literature regarding the use of both exercise and pharmacological stress testing in conjunction with echocardiography in the settings of native and prosthetic mitral and aortic valve disease. We will also discuss the limitations of this modality and touch upon possible future areas of investigation. PMID- 10453299 TI - Stress echocardiography in women. AB - The diagnosis of coronary heart disease in women has been thought to be more difficult than in men, owing to the overall lower prevalence and severity of disease in women, as well as more subtle clinical presentations. Exercise electrocardiography is associated with a high rate of false-positive results. In contrast, exercise and pharmacologic stress echocardiography have been shown to have high sensitivity, specificity, and prognostic value in women, comparable to that obtained in a male population. Although exercise thallium provides high f disease accuracy, due to its cost, availability, and radiation exposure, it may not be the ideal initial test in women. Thus, compared with other modalities, the advantages of stress echocardiography include its lower cost, availability, and high diagnostic accuracy. In the evaluation of women with chest pain, the initial step should involve clinical stratification into low, moderate, or high probability groups based on symptoms, age, and cardiovascular risk factors. In women with atypical chest pain and a low probability of coronary heart disease, further testing should be avoided because any positive result is likely to be falsely positive. In those women with a moderate likelihood of disease, the most efficient and cost-effective strategy includes stress echocardiography as the initial test. This approach avoids the high rate of false-positive results with subsequent unnecessary angiography generated by exercise electrocardiography, as well as minimalizing false-negative results, which would lead to delays and potential increase in morbidity and mortality from untreated coronary heart disease. The optimal strategy for women at high clinical risk may be either exercise echocardiography or cardiac catheterization as the initial test. Although the diagnosis of CAD in women is different than in men, it is not necessarily more difficult. Astute clinical evaluation, in conjunction with judicious use of diagnostic testing, yields excellent results. PMID- 10453300 TI - Cost-effectiveness of stress-echocardiography. AB - Because of constraints on the costs of providing medical care, cardiologists in the future will find themselves challenged to provide care for their patients in the most cost-effective manner possible. Although stress-echocardiography has been shown to compare favorably with other tests in diagnostic accuracy, data on cost-effectiveness are scarce. In this article, general concepts of cost effectiveness as they relate to stress-echocardiography are reviewed and the available literature is summarized. Although definitive data are lacking, there is evidence to suggest that stress-echocardiography may prove to be cost effective in several clinical situations. PMID- 10453301 TI - Choosing the appropriate stress modality. A clinical cardiologist's perspective. AB - The principle application of stress testing is the detection of myocardial ischemia in patients suspected of having coronary artery disease. Stress testing is also used to assess physical exercise capacity in normal controls and patients with various types of heart disease, such as cardiomyopathy and valvular disease. The various choices of stress modalities are discussed in this article. PMID- 10453302 TI - Problems and opportunities for nutrition support practitioners. PMID- 10453303 TI - Principles for formulating parenteral nutrition for neonates. AB - Total parenteral nutrition in humans began with work on neonates, and every year since then there have been important new discoveries. In the past 12 months there have been important new preliminary findings in parenteral nutrition cholestasis, and in bone mineralization. There has been further work on clinical pathways and other aspects of quality assurance, and scientists continue to find new aspects. PMID- 10453304 TI - Recent advances in the placement of tubes for enteral nutrition. AB - Enteral nutrition is the preferred route for nutritional support compared with parenteral nutrition if the gastrointestinal tract is functionally preserved. Long-standing nasogastric or nasoenteric feeding tubes are not well tolerated. Alternative routes are gastrostomy and jejunostomy. Percutaneous endoscopic gastrostomy/jejunostomy or those guided by fluoroscopy, sonography or tomography should be the first choices. Laparoscopy or laparotomy gastrostomy/jejunostomy routes should be reserved for specific situations. Insufflation of the stomach with air or saline solution facilitates the placement of nasoenteric feeding tubes or percutaneous sonographic-guided gastrostomy. The gastrostomy button is a safe and aesthetic alternative, at least in children. Comparison between percutaneous endoscopic gastrostomy and surgical gastrostomy performed either via laparotomy or laparoscopy favours the first in terms of costs and risks. Whenever associated intra-abdominal procedures or anatomic difficulties arise, a laparoscopic or an open access becomes necessary. Complications with feeding tubes are not uncommon and should be promptly recognized and treated. PMID- 10453305 TI - New options for pumps and tubes: progress in enteral feeding techniques and devices. AB - In the past, blenderized tube feeds prepared in the hospital kitchen were delivered by bolus or gravity at intervals of 4-6 h. These methods lack consistent steady flow and many times are not a safe mode of delivery for those at risk of vomiting and aspiration, and cause more patient discomfort than nutritional benefit. More recently, enteral feeding administration techniques and devices have been developed for the delivery of commercially prepared enteral formulas designed for specific disease states. These technological advances have improved enteral feeding practices. PMID- 10453307 TI - Nutrient regulation of gene and protein expression. PMID- 10453306 TI - Technical aspects of trace element supplementation. AB - Routine supplementation of total parenteral nutrition mixtures with the readily available single or combination trace elements products is becoming more widespread. As more is learned about deficiency syndromes and monitoring techniques, so too must we understand more about the physicochemical interactions between individual trace elements and other nutrients, that could ultimately affect bioavailability. Expert pharmaceutical assessment of these complex reactions, that have been demonstrated to occur in solution, becomes increasingly important in order to optimize the efficacy of micronutrient therapy. PMID- 10453308 TI - Endothelial nitric oxide synthase expression and its functionality. AB - Endothelium-derived nitric oxide is a key determinant of blood pressure homeostasis and platelet aggregation, and is synthesized by the endothelial isoform of nitric oxide synthase. In the vascular wall, endothelial nitric oxide synthase is activated by diverse cell surface receptors and by an increase in blood flow. Although initially classified as a constitutive enzyme, many substances and conditions that regulate endothelial nitric oxide synthase gene expression are now described. With the recent discovery of endothelial nitric oxide synthase being compartmentalized in signal-transducing microdomains of the plasma membrane termed caveolae, the past 2 years have mostly witnessed important advances in understanding the control of the functional expression of the enzyme. PMID- 10453309 TI - Amino acid regulation of gene expression. AB - In mammals, the plasma concentration of amino acids is affected by nutritional or pathological conditions. For example, an amino acid profile alteration has been reported as a result of a deficiency of any one of the essential amino acids, a dietary imbalance of amino acids or an insufficient intake of protein. Amino acid availability regulates the expression of several genes involved in the regulation of growth, cellular function or amino acid metabolism. A limitation of several amino acids strongly increases the expression of insulin-like growth factor binding protein CHOP and asparagine synthetase genes. Elevated messenger RNA levels result from both an increase in the rate of transcription and an increase in messenger RNA stability. DNA amino acid response elements have been characterized in the promoter of CHOP and asparagine synthetase genes. The underlying mechanisms of gene regulation by amino acid limitation are not yet completely understood. The results discussed in this review demonstrate that amino acids by themselves can play, in concert with hormones, an important role in the control of gene expression. PMID- 10453310 TI - Nutrient regulation of intestinal gene expression. AB - This review examines recent advances in the dietary modulation of gene expression in the gastrointestinal tract. We have chosen to concentrate on individual genes and examine what is known about their regulation, attempting to link different studies together. PMID- 10453311 TI - Peroxisome proliferator-activated receptors: mediators of a fast food impact on gene regulation. AB - Peroxisome proliferator-activated receptors are nuclear receptors with pleiotropic effects on intra- and extracellular lipid metabolism, glucose homeostasis, inflammation control, and cell proliferation. This review addresses the respective roles of the different peroxisome proliferator-activated receptor isoforms in these different processes. PMID- 10453312 TI - The role of nutrition before and after transplantation. PMID- 10453313 TI - Intestinal transplantation. AB - Intestinal transplantation might become the alternative to definitive parenteral nutrition in patients with permanent intestinal failure. Indeed, recent advances in immunosuppressive treatment and better monitoring and control of acute rejection have brought intestinal transplantation into the realms of standard treatment of intestinal failure. This procedure may be performed in adult or paediatric patients under certain conditions. This short review focuses on the current clinical results and indications for intestinal transplantation and discusses the strategy regarding this challenging procedure. PMID- 10453314 TI - Glutamine in the support of patients following bone marrow transplantation. AB - Bone marrow transplantation is being utilized with increasing frequency in the treatment of patients with malignancy; it is also being applied to the treatment of patients with genetic diseases and as an adjunct to solid organ transplantation. The high dose cytotoxic chemotherapy, often accompanied by total body irradiation, results in severe catabolism, disruption of the gastrointestinal mucosa and marked immunosuppression. A variety of studies show that the supplementation of the amino acid glutamine, by the enteral or parenteral route, as either the free or dipeptide form, appears safe and efficacious in patients undergoing bone marrow transplantation. Further double blind controlled clinical trials of glutamine supplementation in patients undergoing bone marrow transplantation and receiving more contemporary treatment, which often includes the administration of novel combinations of cytoreductive agents and hematopoietic growth factors, are warranted. PMID- 10453315 TI - Haemodialysis and peritoneal dialysis: metabolic alterations and nutritional status. AB - In dialysis patients, malnutrition is an independent factor causing morbidity and mortality. Both inadequate alimentation and metabolic alterations, which involve nitrogen and energy metabolism, contribute to malnutrition. Future research must address the treatment of anorexia and inflammation-induced catabolism, as well as the evaluation of nutritional supplementation techniques and anabolic drugs. PMID- 10453316 TI - Gravity and space flight: effects on nutritional status. AB - The final decade of the millennium has seen an enormous amount of on-orbit life sciences research, including both short- and long-duration flight research. Life sciences dedicated Space Shuttle flights have made intensive research opportunities available to study on the acute adaptation to weightlessness. The NASA/Mir Science Program combined resources of the USA and Russia to provide the first long-duration flight opportunities for the United States since the Skylab program of the early 1970s. Many of the results of these studies are still being evaluated, and in some cases data are still being collected to assess long-term readaptation to gravity after several months in weightlessness. The surge in life sciences research during this decade serves as a preamble to the opportunities to be provided by the latest addition to the Earth-orbiting structures--the International Space Station. PMID- 10453317 TI - Metabolic sequelae of cancers (excluding bone marrow transplantation). AB - The pathogenesis of cancer anorexia/cachexia is still unclear, partly explaining why its treatment remains disappointing. Anorexia plays a central role but cancer cachexia is more complex than chronic starvation. One of the key differences is the preferential mobilization of fat and the sparing of skeletal muscle in simple starvation compared to an equal mobilization of fat and skeletal muscle in cancer patients. An increase in basal energy expenditure also appears to play a contributory role in many patients. Cytokines, essentially but not exclusively tumor necrosis factor-alpha, play an essential pathogenic role and the syndrome can be compared to a low grade chronic inflammatory state. Parenteral nutrition could facilitate the administration of complete doses of chemotherapy or radiotherapy but no significant survival benefit or decrease in treatment-induced toxicity have been demonstrated in prospective randomized trials. The gut should have the preference for nutritional support. Percutaneous endoscopic gastrostomy is used more and more often in patients with a functionally intact gastrointestinal tract, especially in patients with head and neck cancer. Progestational drugs can to some extent stimulate appetite, food intake, energy level, increase weight and decrease the severity of nausea and vomiting. However, pharmacological treatment of cancer cachexia remains disappointing and more trials with anticytokine drugs, anabolic agents or polyunsaturated fatty acids should be conducted. PMID- 10453319 TI - Bibliography. Current world literature. Technical aspects of nutritional support. PMID- 10453318 TI - Liver cirrhosis: rationale and modalities for nutritional support--the European Society of Parenteral and Enteral Nutrition consensus and beyond. AB - Evaluation of nutritional status is a major problem in patients with liver cirrhosis this is due to water retention and the effect of liver function on protein synthesis. Despite problems evaluating the patient, malnutrition has been found to be a common complication in liver cirrhosis and is associated with poorer outcome. Nutritional restrictions, like protein restriction, are no longer recommended in most patients with liver cirrhosis but are considered harmful. An intake of 1 to 1.5 g/kg protein and 25 to 40 kcal/kg body weight a day is recommended (depending on the situation of the patient). If adequate intake cannot be achieved by oral nutrition, stepwise nutritional support with the introduction of an additional late evening meal, sip feeding or tube feeding is recommended. Parenteral nutrition should be used as a second line treatment for acutely ill patients. Data indicate that improvement of nutritional status prior to liver transplantation might reduce complications. PMID- 10453320 TI - Bibliography. Current world literature. Gene and nutrition. PMID- 10453321 TI - Bibliography. Current world literature. Nutrition in wasting diseases. PMID- 10453322 TI - Antioxidants and nutrition support. PMID- 10453323 TI - Amino acids and proteins--new definitions and requirements, hormonal interactions, methodological advances and pitfalls. PMID- 10453324 TI - Counter-regulatory hormones and mechanisms in amino acid metabolism with special reference to the catabolic response in skeletal muscle. AB - There is evidence that both counter-regulatory hormones, in particular glucocorticoids, and cytokines influence amino acid and protein metabolism in skeletal muscle, and that these two groups of regulators interact in the development of muscle catabolism. Glucocorticoids stimulate muscle proteolysis during sepsis and also in other catabolic conditions. In addition, glucocorticoids regulate muscle glutamine metabolism, resulting in increased glutamine release and reduced glutamine concentrations in skeletal muscle. Glucocorticoids inhibit the glutamine transporter in skeletal muscle and stimulate glutamine synthetase activity. Proinflammatory cytokines, in particular tumor necrosis factor and interleukin-1, inhibit muscle amino acid transport by system A, and these cytokine effects are probably indirect. Most of the catabolic effects of tumor necrosis factor in skeletal muscle, including stimulated protein degradation and inhibited amino acid uptake, are mediated by glucocorticoids. PMID- 10453325 TI - Deterrents to the successful clinical use of growth factors that enhance protein anabolism. AB - Participants in competitive sports have demonstrated that the use of growth factors and other anabolic agents enhance human performance, yet physicians are slow to adopt this approach in patients who have a disease-related decrease in strength and activity. Growth factors should be thought of as the next major step forward in providing more efficient and effective nutritional support to catabolic or wasted patients. The obstacles to the use of these agents include: the lack of convincing clinical studies; concerns with safety; anabolic effects and the patient's nutritional status; interaction with diet and route of nutrient administration; the training of health professionals to administer growth factors; and cost. The benefits from these agents should be directly translated into improved patient outcome. To achieve this goal, a uniquely educated group of investigators, working with sophisticated representatives from the pharmaceutical industry, must emerge to construct appropriate protocols and determine desirable endpoints to evaluate the true societal benefits of these agents. PMID- 10453326 TI - Of flux and flooding: the advantages and problems of different isotopic methods for quantifying protein turnover in vivo: I. Methods based on the dilution of a tracer. AB - The advantages and problems, both practical and theoretical, of isotope dilution approaches to the determination of whole-body and tissue protein turnover are discussed. It was concluded that: (1) measurements made on the basis of the labelling of plasma and breath are well suited to the measurement of body amino acid oxidation and balance, but because of the problem of inhomogeneity of the body amino acid pools, this approach generally underestimates protein turnover; (2) in investigations of nutritional effects on whole-body amino acid turnover, closer attention should be paid to first-pass splanchnic amino acid metabolism; (3) the trans-organ tracer balance method, particularly if combined with the measurement of tissue amino acid labelling, is a potentially useful approach to the simultaneous and dynamic measurement of both protein synthesis and degradation; (4) leucine may be the most generally useful label for tracer level studies of both whole-body and muscle protein synthesis, as recent studies have shown quite close isotopic equilibrium between muscle-free and tRNA-bound leucine pools. PMID- 10453327 TI - Discrepancy between muscle and whole body protein turnover. AB - To understand the mechanisms of muscle wasting, muscle protein turnover is measured. Several techniques are available, including whole body protein turnover measurements and incorporation of tracer into mixed muscle protein. In this brief review, we will focus on these techniques. We conclude that whole body measurements cannot be used as a parameter of muscle protein metabolism. It appears that whole body measurements mainly represent splanchnic protein metabolism. Muscle protein synthesis preferably is measured in individual proteins or protein subfractions. PMID- 10453328 TI - The use of alpha-ketoglutarate salts in clinical nutrition and metabolic care. AB - Theoretically, alpha-ketoglutarate is a precursor of glutamine, a fact that may be of importance given the key regulatory properties of this amino acid. Although the literature suggests that glutamine synthesis accounts only for a marginal part of the disposal of exogenously supplied alpha-ketoglutarate, administered alpha-ketoglutarate has a potent 'sparing' effect on endogenous glutamine pools. When alpha-ketoglutarate is supplied as an ornithine salt, a synergistic effect of the two parts of the molecule increases the synthesis of glutamine or the 'sparing' of endogenous glutamine pools. In addition, alpha-ketoglutarate in combination with ornithine dramatically increases the synthesis of arginine, proline and polyamines, which also play key roles in metabolic adaptation to trauma. The recent literature suggests that the administration of alpha ketoglutarate in combination with ornithine improves gut morphology and functions, counteracts trauma-induced dysimmunity and exerts anabolic/anticatabolic actions on protein metabolism. PMID- 10453329 TI - Adult human amino acid requirements. AB - This brief review is concerned with the status of our current understanding of the quantitative needs for dietary amino acids in healthy adults. The nutritional significance of the conditionally indispensable amino acids is assessed and the requirements for the indispensable amino acids are discussed. Recent research involving the use of the tracer balance approach is reviewed, and the importance of resolving the current uncertainties regarding adult human amino acid needs is highlighted. PMID- 10453330 TI - Amino acid metabolism in liver disease. AB - The impairment of transsulphuration during methionine degradation in hepatic failure can be counteracted by treatment with S-adenosylmethionine. Regarding the pathogenesis of hepatic encephalopathy, no convincing evidence exists for tryptophan, glutamine or glutamate being involved. Portal-systemic shunting induced hyperammonaemia may reduce plasma branched-chain amino acids. The glucose effect on urea synthesis does not exist in cirrhosis. PMID- 10453331 TI - Perioperative nutrition. PMID- 10453332 TI - Indirect calorimetry: can this technology impact patient outcome? AB - This review of 23 papers involving indirect calorimetry published over the past 18 months shows how our understanding of the metabolic response to injury has changed, highlights the problems introduced by use of predictive equations and alterations in indirect calorimetry testing protocol, and emphasizes the need to monitor cumulative energy balance by comparing daily caloric intake to energy expenditure. PMID- 10453333 TI - Insulin resistance: a marker of surgical stress. AB - Elective surgery causes a marked, transient reduction in insulin sensitivity. The degree of the reduction is related to the magnitude of the operation. The type and duration of surgery performed, perioperative blood loss, and also the degree of postoperative insulin resistance have significant influences on the length of hospital stay. A novel approach to minimize insulin resistance after surgery is being presented and suggests that simply pretreating the elective surgical patient with sufficient amounts of carbohydrates instead of fasting can significantly reduce postoperative insulin resistance. It is not clear which mediators are the most important for the development of insulin resistance after surgery. Nevertheless, marked insulin resistance can develop after elective surgery without concomitant elevations in cortisol, catecholamines or glucagon. The main sites for insulin resistance seem to be extrahepatic tissues, probably skeletal muscle, where preliminary data suggest that the glucose transporting system is involved. PMID- 10453334 TI - Perioperative hyperglycemia, infection or risk? AB - This review covers the recent studies that have served to further our understanding of the nature of the relationship between perioperative hyperglycemia and nosocomial infection. On the one hand hyperglycemia can be a consequence of the systemic inflammatory response, and can serve as a marker of the severity of stress and the degree of immunocompetence resulting from infection or injury. Strong evidence is, however, emerging that hyperglycemia in the perioperative period can also be a significant risk factor for the development of nosocomial infection. PMID- 10453335 TI - Gut microenvironment and immune function. AB - Overreaction of the acute phase response is responsible for the two major complications to surgery, sepsis and thrombosis, but also most likely for the leading sequela to surgery, adhesion formation. The gastrointestinal tract, especially the colon, is a major player in the acute phase response and responsible for important immune functions with important interactions between the commensal flora, mucosal cells and the mucosa/gut associated lymphoid tissues. These responses can effectively be modulated by enteral nutrition, provided it is properly composed and administered. There is increasing evidence that the important clinical effects sometimes observed in enteral nutrition are more related to immunostimulatory effects than to reduction in microbial translocation. It is suggested that in order to be effective enteral nutrition should be instituted if possible before the operation, but always at least immediately after. Furthermore, much supports that the formula given should contain what has been called colonic food, e.g. plant fibres, and have a low content of saturated fat. Use of antibiotics with deleterious effects on the commensal flora should also be limited as much as possible. Lack of compliance with these requests seems to explain the lack of consistency in clinical experience of enteral nutrition, when tried in connection with trauma and clinical surgery. PMID- 10453337 TI - Bibliography. Current world literature. Protein metabolism and therapy. PMID- 10453336 TI - Nutritional support and aging in preoperative nutrition. AB - In the past 20 years, an increased interest geriatric nutrition has induced researchers to document the distribution and magnitude of nutritional problems in the elderly population. It has been observed that the prevalence of malnutrition is greatly affected by the general health status and autonomy of the elderly. Among free-living healthy elderly persons, the prevalence of protein-caloric undernutrition is low. As health and functional capacities deteriorate with age, however, the prevalence increases dramatically to 30-65% of those in home care, nursing homes or in hospital. Formal nutritional assessment has typically been absent from most published programmes of geriatric evaluation and comprehensive geriatric assessment. This is frequently because of the lack of a specific validated tool to assess nutritional status in older persons and, at least partly, to explain this phenomenon. The Mini Nutritional Assessment was developed and validated on large representative samples of elderly persons to address these specific issues. Recent experimental studies have shown that advanced malnutrition is much more difficult to treat in the elderly than in younger adults. Trials of nutritional support using oral supplements or enteral tube feeding have shown improved outcome in those identified as malnourished on admission to hospital. PMID- 10453338 TI - Human obesity: a sufficient cause. AB - It is sometimes useful to group the causes for any biological event into those that are necessary and those that are sufficient. Our current, partial understanding of human obesity has come through a focus on readily accessible necessary causes. The sufficient causes are likely to be more complex, dealing with the integration of developmental events, behavior and complex biochemical systems. PMID- 10453339 TI - Dietary lipids: more than just a source of calories. PMID- 10453340 TI - Fatty acids and endothelial cell function: regulation of adhesion molecule and redox enzyme expression. AB - Different types of dietary fatty acids have distinct, and sometimes opposing, effects on endothelial cell function. Long-chain n-3 polyunsaturated fatty acids from fish oil (eicosapentaenoic-acid-20:5n-3 and docosahexaenoic-acid-22:6n-3) attenuate cytokine-induced adhesion molecule expression (mRNA and protein) while the n-6 polyunsaturated fatty acid arachidonic acid (eicosatetraenoic-acid-20:4n 6) either has no effect, or elicits as up- or down-regulation. Conjugated linoleic acids derived from 18:2n-6, linoleic acid also reduced the expression of adhesion molecules. These polyunsaturated fatty acids also induced redox enzyme expression (mRNA and protein) in endothelial cells and modulation of redox sensitive transcription factors (e.g. nuclear factor kappa B, activated protein 1) which regulate the gene expression of adhesion molecules, redox enzymes and various stress proteins. The induction of redox enzyme expression by n-3 polyunsaturated fatty acids and conjugated linoleic acids could explain their inhibitory effects on gene transcription and adhesion molecule protein expression. Individual adhesion molecule genes and transcription factors appear to differ in their responsiveness to various oxidant stimuli and non-redox, more direct regulatory mechanisms (e.g. peroxisome proliferator activation receptor activation) might also be involved in their regulation. PMID- 10453341 TI - Lipid treatment of inflammatory bowel disease. AB - The aetiology of inflammatory bowel disease remains unclear. Local mediators such as arachidonic acid metabolites and peptide mediators (cytokines) appear to contribute to the disease process. The successful administration of neutralizing antibodies against TNF-alpha has confirmed a pathophysiological role for this cytokine in Crohn's disease. Established therapy of inflammatory bowel disease with 5-aminosalicylic acid compounds has been shown to reduce local leukotriene B4 formation by inhibiting lipoxygenases. This therapeutic mechanism formed one rationale for examining the effect of n-3 fatty acids, which also inhibit leukotriene B4 formation, on the course of these diseases. In the first study, published in 1989, we found no beneficial effects of n-3 fatty acids in patients with Crohn's disease; however, there was clinical improvement, just falling short of significance, in patients with ulcerative colitis. Since then two uncontrolled and five controlled studies have further investigated the therapeutic effect of n 3 fatty acids in patients with ulcerative colitis. The size of the patient population in the controlled studies ranged from 10 to 96 patients in the largest study. Two of these studies showed a significant improvement in clinical activity and a steroid-sparing effect, respectively. Another study found only a trend towards improvement and one trial, which also included a treatment group receiving evening primrose oil, found no beneficial effect in the 16 patients receiving n-3 fatty acids. A large, 2 year trial of n-3 fatty acids in patients with ulcerative colitis off steroids, which was recently completed at the Universities of Munich and Mainz, showed a delay of the first episode of relapse, but no reduction in the cumulative relapse rate at 2 years. Controversial results have been published for Crohn's disease. A new enteric-coated formulation reportedly increased the proportion of patients in remission where as another trial using a conventional preparation found no significant effect. PMID- 10453342 TI - n-3 polyunsaturated fatty acids and cancer. AB - n-3 Polyunsaturated fatty acids are promising molecules in cancer prevention and the potentiation of cancer treatment. Recent studies have highlighted the importance of their interactions with other food components. Their effects on tumor growth depend upon background levels of n-6 polyunsaturated fatty acids and antioxidants, and this could account for previously inconsistent results in experimental carcinogenesis. Recognition of the role of lipoperoxidation in the anti-tumor effects of polyunsaturated fatty acids, which is apparent in a variety of in-vitro or in-vivo systems, has been a major advance in the field. Consequently, n-3 polyunsaturated fatty acids appear to be excellent substrates for lipid peroxidation in situations where an oxidative stress is involved, such as in the action of several cytotoxic agents in the treatment of cancer. PMID- 10453343 TI - Fatty acids: links between genes involved in fatty acid and cholesterol metabolism. AB - Fatty acids are a major constituent of dietary fats and form an integral part of the cellular membrane and lipoproteins. The gene regulatory potential of fatty acids has long been recognized, but the precise regulatory mechanisms are unknown. The regulatory ability of fatty acids on the expression of a number of genes together with potential mechanisms and pathways of regulation are reviewed. In this review, we emphasize a key aspect of regulation mediated by the sterol regulatory element binding-protein, and its effects on sterol regulatory elements. PMID- 10453344 TI - An update on the pathways of polyunsaturated fatty acid metabolism. AB - The biosynthesis of 4, 7, 10, 13, 16-22:5 and 4, 7, 10, 13, 16, 19-22:6 from dietary linoleate and linolenate, respectively, does not totally take place in the endoplasmic reticulum but does require the participation of enzymes in the endoplasmic reticulum and peroxisomes. The absence of an endoplasmic reticulum associated acyl-CoA-dependent delta 4 desaturase also requires the controlled movement of 22- and 24-carbon polyunsaturated fatty acids between the endoplasmic reticulum and peroxisomes. PMID- 10453345 TI - Clinical use of lipid emulsions. AB - Substantial progress has been made in the understanding of the metabolism of intravenous lipid emulsions and the delivery of their various components to specific tissues or cells. Lipid emulsions should be considered not only as a means of providing energy substrates but also specific compounds that participate in the regulation of key metabolic functions. Such improved knowledge should find applications in the metabolic care of different types of patients. PMID- 10453346 TI - Lipids in enteral nutrition. AB - Lipids in enteral nutrition facilitate the presentation of a high energy source with low osmotic impact. Focus has shifted from macronutrients towards the inclusion of special nutrients and growth factors. Recent advances in the design of triacylglycerol lipids with specific structures facilitate the absorption of essential fatty acids of the n-3 series, which provide specific benefits with respect to tissue repair and to the immune system. Enteric formulations containing n-3 lipids are proving to be of value in sustaining seriously ill patients. Information from well-controlled trials is generally consistent in establishing the benefits of formulations containing n-3 lipids. PMID- 10453347 TI - Pathways of muscle protein breakdown in injury and sepsis. AB - The purpose of this article is to review evidence that the ubiquitin-proteasome proteolytic pathway plays an important role in injury- and sepsis-induced muscle catabolism. Such evidence includes upregulated gene expression of several of the components of the ubiquitin-proteasome pathway as well as energy-dependency of the injury- and sepsis-induced muscle protein breakdown. Although the ubiquitin proteasome pathway is the predominant mechanism of muscle breakdown in various catabolic conditions, other proteolytic mechanisms, in particular calcium dependent, calpain-mediated protein degradation, probably participate as well. PMID- 10453348 TI - Autoregulation of glucose production in health and disease. AB - Plasma glucose concentration is a major regulator of glucose production in experimental animals and in humans. Hyperglycemia inhibits and hypoglycemia stimulates glucose production through mechanisms which are, in part, independent of changes in glucoregulatory hormones. Glucose production does not change when gluconeogenesis is acutely inhibited or stimulated. These observations indicate an autoregulatory process by which hepatic glucose production is altered as a direct consequence of changes in glycemia. Data obtained in patients with glucokinase gene mutations indicate that glucokinase may play a role in this regulatory process. Hyperglycemia in type 2 diabetic patients and in critically ill patients may result from a blunted effect of hyperglycemia on glucose production. PMID- 10453349 TI - Energy metabolism in critically ill patients: lactate is a major oxidizable substrate. AB - The potential role of an energy defect in acute diseases is still in the centre of the pathophysiological understanding of such states and therefore of our attempts to limit or to reverse the possible deleterious consequences of such defect. In fact several recent experimental works have shown that instead of being a negative consequence, the lactate production and the related metabolic acidosis due to the stimulation of anaerobic ATP-production pathway is rather a protective adapted response. PMID- 10453350 TI - Anabolic hormones in critically ill patients. AB - Critical illness and severe injury induce a number of neuroendocrine changes that initially enhance survival. However, for skeletal muscle, the hormonal environment is such that accelerated catabolism predominates. The prevailing hypercortisolemia is exacerbated by a resistance to or decreased production of anabolic hormones, or both. The resulting loss of lean body mass compromises the patient's ability to handle acute complications and impairs efforts towards rehabilitation. For these reasons, efforts have been focused upon the restoration of the anabolic influence. Recent studies continue to demonstrate that the amelioration of muscle catabolism is possible with hormonal management. Emerging technologies and conservative administration regimens hold promise for the development of the clinical ability to restore anabolic influence, while minimizing undesirable side effects. PMID- 10453352 TI - Bibliography. Current world literature. Lipid metabolism and therapy. PMID- 10453351 TI - Glutamine: establishing clinical indications. AB - Glutamine, a non-essential amino acid, is abundant in the human body and in the food we eat; it is mainly produced in skeletal muscle and acts as a major inter organ nitrogen and carbon transporter. Its importance to the metabolism is evident during stress, when it becomes a conditionally essential amino acid when endogenous supply fails to meet increased demands. During the past 2 years, an increased understanding has been gained into the role of glutamine in metabolism. A number of new clinical studies of glutamine supplementation have shown interesting outcomes that should be influencing clinical decision-making. PMID- 10453353 TI - Bibliography. Current world literature. Nutrition in the intensive care unit. PMID- 10453354 TI - Molecular control of immune/inflammatory responses: interactions between nuclear factor-kappa B and steroid receptor-signaling pathways. PMID- 10453355 TI - The corticotropin-release inhibitory factor hypothesis: a review of the evidence for the existence of inhibitory as well as stimulatory hypophysiotropic regulation of adrenocorticotropin secretion and biosynthesis. PMID- 10453356 TI - The cyclins and cyclin-dependent kinase inhibitors in hormonal regulation of proliferation and differentiation. PMID- 10453357 TI - The insulin-related ovarian regulatory system in health and disease. PMID- 10453358 TI - Citation for the 1999 Fred Conrad Koch Award of the Endocrine Society to Dr. Ronald M. Evans and Dr. Michael G. Rosenfeld. PMID- 10453359 TI - Citation for the 1999 Ernst Oppenheimer Award of The Endocrine Society to Dr. David W. Russell. PMID- 10453360 TI - Citation for the 1999 Robert H. Williams Distinguished Leadership Award of the Endocrine Society to Dr. Joel F. Habener. PMID- 10453361 TI - Citation for the 1999 Edwin B. Astwood Lecture Award of the Endocrine Society to Dr. David D. Moore. PMID- 10453362 TI - Citation for the 1999 Monsanto Clinical Investigator Lecture Award of the Endocrine Society to Dr. Stephen J. Marx. PMID- 10453363 TI - Citation for the 1999 Gerald D. Aurbach Lecture Award of the Endocrine Society to Dr. James E. Darnell. PMID- 10453364 TI - Citation for the 1999 Sidney H. Ingbar Distinguished Service Award of the Endocrine Society to Dr. M. Susan Smith. PMID- 10453365 TI - Citation for the 1999 Roy O. Greep Lecture Award of the Endocrine Society to Dr. Ernst Knobil. PMID- 10453366 TI - Citation for the 1999 Distinguished Educator Award of the Endocrine Society to Dr. Daniel D. Federman. PMID- 10453367 TI - Citation for the 1999 Distinguished Physician Award of the Endocrine Society to Dr. Henry G. Burger. PMID- 10453368 TI - Citation for the 1999 Richard E. Weitzman Award of The Endocrine Society to Dr. Jacques Simard. PMID- 10453369 TI - Prospective evaluation of transabdominal bowel sonography in the diagnosis of intestinal obstruction in Crohn's disease: comparison with plain abdominal film and small bowel enteroclysis. AB - Transabdominal ultrasonography is a noninvasive, radiation-free method that is well tolerated by patients with acute abdominal symptoms. The aim of this study was to investigate the validity of transabdominal ultrasonography, compared with plain abdominal film and small bowel enteroclysis in the diagnosis of small bowel obstruction in patients with Crohn's disease (CD). Forty-four patients with CD ileitis or ileocolitis and acute obstructive symptoms who received ultrasonography and plain abdominal film at hospital admission were evaluated by small bowel enema before dismissal or surgery and were considered for statistical analysis. Small bowel obstruction was diagnosed by ultrasonography in 23 of 44 patients (52%), by plain abdominal film in 26 of 44 patients (59%), and by small bowel enema in 28 of 44 patients (64%); the diagnostic accuracy of plain abdominal film and ultrasonography compared with small bowel enema was 73% and 89%, respectively. Ultrasonography proved to be highly specific (100%) with no false positive results. Surgery performed in 25 of 44 patients for symptoms refractory to medical treatment confirmed the high diagnostic value of ultrasonography. The result of this study indicates that transabdominal ultrasonography is accurate and highly specific in the diagnosis of small bowel obstruction and can be considered a valuable first choice examination in CD patients with obstructive symptoms. PMID- 10453370 TI - Presenting symptoms and diagnostic lag in children with inflammatory bowel disease. AB - Presenting symptoms and their duration may affect the time that elapses prior to definitive diagnosis of inflammatory bowel disease (IBD). This study was undertaken to determine the mean duration of presenting symptoms and diagnostic lag in children with IBD. The medical records of all patients less than 19 years of age diagnosed with IBD at the pediatric gastroenterology clinic of Children's Hospital of Wisconsin between 1990-1995 were reviewed. The age at diagnosis, gender, presenting symptoms and duration, disease location, and diagnostic lag were analyzed. There were 91 children (49 male) diagnosed with IBD. Crohn's disease (CD) was diagnosed in 58, ulcerative colitis (UC) in 24, and indeterminate colitis in 9. The mean ages at diagnosis were 11.4 years for CD, 9.7 years for UC, and 7.8 years for indeterminate colitis. The most frequent presenting symptoms were abdominal pain, diarrhea, hematochezia, and weight loss. The average lag in diagnosis of CD was 7.1 months, which varied by disease location: small intestine 10.5 months, ileocolonic 7.5 months, and colonic 6.4 months. The average lag in diagnosis was 6.7 months for UC and 14 months for indeterminate colitis. Children presenting with growth failure had the longest diagnostic lag. (a) The elapsed time between symptom onset and the diagnosis of CD has decreased. (b) The diagnostic lag in CD decreases with distal colonic involvement. (c) Following onset of symptoms UC was diagnosed only slightly more rapidly than CD. PMID- 10453371 TI - Bone mineral density in children and young adults with Crohn's disease. AB - Reduced bone mineral density (BMD) has been reported in adults with Crohn's disease (CD). Less is known about abnormal BMD in children and young adults with CD. The aims of this study are to determine the prevalence of low BMD and to evaluate the effect of growth and pubertal development on BMD in children and young adults with CD. One hundred-nineteen patients with CD underwent dual-energy X-ray absorptiometry (DXA) to determine BMD. Anthropometry and pubertal development were measured. Bone age was measured only in patients older than 8 years of age and who had not grown in height during the last year. One hundred nineteen patients (72 male, 47 female) were evaluated. Seventy percent of patients had BMD z-scores < or = -1.0 and 32% had z-scores < or = -2.0. Weight and height z-scores were significantly associated with BMD z-scores. BMD z-scores based on bone age and on chronological age were highly correlated, except when the chronological age BMD z-score was < or = -2.0. BMD z-score was significantly different between males and females for the group (-1.75 +/- 1.06 vs. -1.08 +/- 1.00), respectively. Children and young adults with CD have a high prevalence of low BMD and routine evaluation by DXA is indicated. In patients with a chronological age-based BMD z-score < or = -2.0, a bone age-based BMD should be considered. PMID- 10453372 TI - Plasma and rectal adenosine in inflammatory bowel disease: effect of methotrexate. AB - In animal models, the antiinflammatory mechanism of action of methotrexate has been attributed to elevation of the extracellular concentration of the endogenous autocoid, adenosine. Our goal was to determine if methotrexate elevates adenosine concentrations in plasma and at the site of disease in patients with inflammatory bowel disease. In 10 patients with Crohn's disease or ulcerative colitis, rectal adenosine and plasma adenosine concentrations were measured before and immediately after a subcutaneous injection of methotrexate, 15 or 25 mg. The mean predose rectal adenosine concentration of 2.4 mumol/l was not significantly different from the postdose concentration of 2.1 mumol/l, p = 0.17, (paired two tailed t test). Rectal adenosine concentration tended to correlate with rectal endoscopic disease activity, r = 0.59, p = 0.067 (Spearman rank order correlation). After methotrexate injection, neither the mean daily plasma adenosine concentration, nor the plasma adenosine at any individual time point, were significantly different from preinjection values. In patients with inflammatory bowel disease, an injection of methotrexate in the clinically effective dose range does not raise rectal or plasma adenosine concentrations. A role for adenosine as the mediator of the antiinflammatory action of methotrexate is not supported. PMID- 10453373 TI - Transdermal nicotine decreases mucosal IL-8 expression but has no effect on mucin gene expression in ulcerative colitis. AB - Our goal was to determine the effect of transdermal nicotine on cytokine and mucin gene transcription in ulcerative colitis (UC). Sixty-four nonsmoking patients with active UC were randomly assigned to transdermal nicotine (maximum dose 22 mg/day) or placebo for 4 weeks. Clinical assessment and colonic mucosal biopsies were obtained at entry and after 4 weeks. Inflammatory and immunoregulatory cytokines were assessed by qualitative reverse transcriptase polymerase chain reaction (RT-PCR). Based on this initial screen. IL-8 mRNA levels were measured by RT-competitive PCR. MUC1, MUC2, MUC3, MUC4, MUC5AC, MUC5B, and MUC6 mRNA concentrations were measured by quantitative dot blot analysis. Cytokine mRNA expression, except for IL-8, was similar in all patients. IL-8 mRNA levels were significantly decreased in the colonic mucosa of nicotine treated patients who improved (p = 0.04). IL-8 mRNA values were similar before and after treatment in nonresponding nicotine-treated patients and in all placebo treated patients. Mucin gene expression was similar in all patient groups. Beneficial effects of transdermal nicotine in active UC may result from decrease of IL-8 expression at the transcriptional level. Transdermal nicotine has no effect on mucin gene transcription. PMID- 10453374 TI - In the X-ray file. PMID- 10453375 TI - Lack of support for a common etiology in Johne's disease of animals and Crohn's disease in humans. AB - The superficial similarity of Johne's disease to Crohn's disease led to the hypothesis that, like the former. Crohn's disease was caused by Mycobacterium paratuberculosis. Detailed pathologic comparisons, however, reveal little similarity between these two entities, including the lack of important extraintestinal manifestations. Attempts to recover M. paratuberculosis by culture have only rarely succeeded and the significance of spheroplasts that appear more frequently on culture is seriously in question. Five immunocytochemistry studies have failed to find mycobacterial antigens in diseased tissues and the five most recent polymerase chain reaction (PCR) attempts to find genomic evidence of M. paratuberculosis were uniformly negative. Numerous serologic studies failed to demonstrate antibody to M. paratuberculosis and attempts to show cell-mediated immunity were also unrewarding. Inoculation of numerous experimental animals with Crohn's disease tissue has failed to induce Johne's disease, and inoculation of various animal species with M. paratuberculosis has equally failed to result in Crohn's disease. Controlled studies of the treatment of Crohn's disease with antimycobacterial agents have generally resulted in no improvement, and most studies that have shown a positive response are either uncontrolled or include broad-spectrum antibiotics that may be acting on pathogens other than mycobacteria. Finally, although Johne's disease is common in farm animals, and infected animals shed M. paratuberculosis in large numbers, no record of zoonotic transmission has been recorded. PMID- 10453376 TI - Bone mineral metabolism in pediatric inflammatory bowel disease. AB - The development of reliable techniques to measure bone densitometry and evolving effective drug treatment have kindled great interest in the diagnosis and treatment of osteoporosis in adults with inflammatory bowel disease. A number of studies have examined the prevalence of abnormal bone mineral metabolism in children and adolescents. Studies, conducted over the past decade, indicate a greater likelihood of clinically significant problems in Crohn's disease than in ulcerative colitis. Corticosteroids have been proven to impair bone mineral status. It is increasingly clear that inflammation and other factors play a bigger role than malabsorbtion of minerals or vitamin D in most patients. As the use of the bisphonate class of drugs is limited in pediatric patients, there is a need to emphasize the role of diet and exercise in children and teenagers, particularly in those affected by inflammatory bowel disease. PMID- 10453377 TI - Metabolic bone disease in adults with inflammatory bowel disease. AB - Bone loss is seen frequently in those with inflammatory bowel disease (IBD). It appears to be more common in those with Crohn's disease than in those with ulcerative colitis. Corticosteroids play an important role in the development of osteoporosis. The prevalence of osteoporosis in IBD, the effects of corticosteroids on bone metabolism, and treatment options are reviewed. PMID- 10453379 TI - An approach to perirectal disease in inflammatory bowel disease. AB - The management of perirectal disease in association with IBD requires an individual approach to the patient with combined medical and surgical therapy where appropriate. No matter what therapy is used, the main goal is to alleviate symptoms and cure the problem where possible. Before undertaking management it is crucial to delineate the extent not only of the local disease but the total extent of the problem in the gastrointestinal tract. This will obviously influence the way in which the local disease is managed. What has been presented is a practical, common-sense approach to the management of what is often an extremely difficult problem. PMID- 10453378 TI - Bones and Crohn's: problems and solutions. AB - Inflammatory bowel disease may manifest in various extra intestinal manifestations. Osteopenia and various arthropathies may be debilitating. These may be related to the disease itself, patient genetics, lifestyle, or disease treatment. Calcium and vitamin D malabsorption, vitamin K deficiency, malnutrition, corticosteroid and other immunosuppressive medications, smoking, lack of exercise and postmenopausal state may all play important roles. Treatment may be undertaken to correct nutrient deficiencies, inhibit bone resorption and increase bone formation. PMID- 10453380 TI - MRI scanning in perianal Crohn's disease: an important diagnostic adjunct. PMID- 10453381 TI - Replacement of cinefilm with a digital archive and review network. AB - The replacement of cinefilm with digital cardiac angiography has accelerated significantly in recent years with the development and widespread adoption of the DICOM standard for interchange of procedure image data. While the acceptance of the Compact Disk-Recordable (CD-R) media has provided the opportunity for numerous laboratories to eliminate cinefilm in all its functions, the task is more complex for larger catheterization laboratories which require the use of high speed networks and automated libraries. The Cardiac Laboratory at the Mayo Clinic has implemented the final stages of a large-scale network and archive which meets a large institution's demanding requirements for access, speed, and storage for high volumes of digital cardiac angiographic images. Initial experience with the system demonstrates that this approach will be a successful one for the elimination of cinefilm and the employment of digital imaging and networking technology. In addition, direct access to digital image records will also facilitate the integration of image data with other clinical information acquired and stored in a digital format. PMID- 10453382 TI - Integration of haemodynamic and electrocardiographic waveform data with DICOM images. AB - The cardiac catheterization procedure involves acquisition of haemodynamic and electrocardiographic waveforms, as well as X-ray angiographic images. The DICOM standards is being extended to address robust interchange of this waveform data together with the image data, so as to present an integrated data set for the procedure. The proposed waveform information object definition is one of the new generation of DICOM extensions, and uses controlled vocabularies and coded entry sequences to represent clinically significant aspects of the data acquisition. This enables a generic waveform object to be specialized for disparate uses, e.g., haemodynamics, electrocardiograms, phonocardiograms, and electrophysiological data. PMID- 10453383 TI - Clinical importance of the DICOM structured reporting standard. AB - The purpose of the DICOM Structured Reporting (SR) specification is to improve the documentation of diagnostic images and waveforms. The specification supports the interchange of expressive compound reports in which the critical features shown by images and waveforms can be denoted unambiguously by the observer, indexed, and retrieved selectively by subsequent reviewers. Findings may be expressed by the observer as: 1) text, codes, numeric measurements: 2) computer generated coordinates of specific regions of interest within images or waveforms; or 3) reference to comparison images, sound, waveforms, curves, and previous report information. The observational and historical findings recorded by the observer may include any evidence referenced as part of an interpretation procedure. Thus, DICOM SR supports not only the reporting of diagnostic observations, but the capability to document fully the evidence that evoked the observations. This capability provides significant new opportunities for large scale collection of structured data for clinical research, training, and outcomes assessment as a routine by-product of diagnostic image and waveform interpretation and facilitates the pooling of structured data for multi-center clinical trials and evaluations. PMID- 10453384 TI - The DICOM review stations: are they truly different? AB - Digital acquisition systems are widely used nowadays. The digitization of the cath lab environment is now directed towards the change in the exchange media from an analog (i.e. cinefilm) to a digital (i.e. CD-R) medium. An important consequence of this development is the need for another type of review system. This article focuses on the replacement of the cineprojector by a digital equivalent: the DICOM review station. Since the technologies differ fundamentally, the DICOM review station has very little in common with its analog predecessor. This article explains the basics of DICOM review stations. Different approaches, both in hardware and in software, are possible, each showing their advantages and drawbacks. The impact of choices made by the industry will be illustrated by a number of commercially available DICOM review stations. Further, the article discusses the topics of diagnostic image quality and performance. In our opinion, these are the major topics when it comes to an objective comparison of the capabilities of DICOM review stations. The relation of these subjects with design choices in terms of hardware and software are discussed. PMID- 10453385 TI - Digital networking and archiving with ACOM T.O.P. AB - The international DICOM 3 standard for a single patient record exchange media (CD R) provided the necessary definition for digital recording and distribution of cardiac angiographic patient data. The medical industry developed archive and review stations that allow the replacement of the 35 mm cine film. SIEMENS launched ACOM. T.O.P. in late 1995. The benefits and technical solution are presented in this paper. Today, the solution for networked real-time image data distribution within the hospital has become reality. Further integration of hemodynamic data, connectivity of hospital information systems, easy access and distribution of such high volume data from and to remote locations is well under way. ACOM. net is used as an example for the realization of commercial products to fulfill the demands of cardiologist and hospital administrators. PMID- 10453386 TI - The status of the GE approach to the digital catheterization laboratory. AB - The digital cardiac catheterization laboratory has become a reality. The cardiac catheterization community has embraced the DICOM standard as the vehicle for filmless archiving of cardiac angiographic studies. General Electric Medical Systems (GE) provides a completely scaleable offering for filmless archiving with the GEMnet product line. The GEMnet philosophy: practicality, scaleability, reliability and upgradeability. Practicality is conforming to standards. Standards in imaging data format and communications. Scaleability of a system configuration maximizes the dollar spent to match the right system to the patient volume of a department. Reliability is a fault tolerant design, redundancy of data to insure system up-time and advanced system diagnostics to speed service time. Upgradeability is keeping pace with the evoluation of new technologies to take advantage of increases in performance and reductions in price. PMID- 10453387 TI - The Kodak approach to digital cardiovascular imaging laboratories. AB - Cine film had been a global standard in the cardiac catheterization lab for the acquisition, archive, review and exchange for decades. The advent of digital imaging in the cath lab heightened the desire to store cath procedure images in a digital format. The issues related to the development of a digital exchange standard, the data capacities and data rates, lossless and lossy compression, and techniques to obtain digital image data from the lab are discussed. In addition, Kodak's approach to cine film replacement with digital systems is described. PMID- 10453388 TI - The Camtronics experience with the filmless digital catheterization laboratory. AB - Systems based on the Archium Digital Cardiac System architecture are providing filmless operation for cardiac catheterization departments in over 80 institutions today. Filmless operation provides direct cost savings from the elimination of cine film as well as its development and management. In addition to these savings, benefits are being realized from productivity associated with changes in the workflow in the department. Image quality and processing capability consistent with the image quality and processing available in the cath lab have proven to be key components in changing workflow and improving efficiency. Solutions are now available which can deliver this level of performance for most departments including multiple lab departments with cath labs from different manufacturers. With Archium, physician productivity can be enhanced with the immediate availability of studies outside the lab and the ability to consult online. Cath lab turnover can be improved significantly. Staff productivity is realized from improved image management as well. The Archium's modular architecture has already accommodated system evolution without obsolescence of existing systems. PMID- 10453389 TI - Integrated image storage solution for the Cath department. AB - Contemporary Image Storage systems for the Catheterization department manage and distribute digital cardiac images according to the "cine-film" paradigm. The images are digital, but the applications have not changed much. This situation will change in the near future. New systems are being developed to store additional (clinical related) data with X-ray Angiographic (XA) Images. Furthermore, the image storage domains are no longer an island in the hospital infrastructure. Efficiency requires the availability of images with other data at the various "point of care" locations. This in turn raises requirements and expectations about the standards in the area of application interoperability, since no single vendor can supply the complete solution. Recent DICOM (Digital Imaging and Communications in Medicine) standardization activities play an important role in extending the current scope of image oriented storage solutions towards a more integrated imaging and information (clinical) folder for the Cath department. The paper will address the following issues: New requirements on "self-contained" Image Storage solutions for the Cath lab. How to deal with the demand for interdepartmental communication using upcoming (new) DICOM standards and HL7 (Health Level Seven) in this area. The increasing influence of computer technology, replacing vendor-specific solutions by general-accepted standards from the Information Technology (IT) world. A step-wise approach to come to an integrated clinical (patient) folder with inherent capabilities for data interchange with other Cardiology departments and the hospitals information infrastructure. PMID- 10453390 TI - How to standardize vasomotor tone in serial studies based on quantitation of coronary dimensions? AB - In patients with coronary artery disease including those after coronary bypass graft operation and heart transplantation intervention studies based on serial quantitative coronary angiography, in part combined with intravascular ultrasound, are of increasing relevance. Since vasomotor tone of epicardial coronary arteries is influenced by a variety of factors, angiographic follow-up studies require standardization of coronary tone by induction of maximal dilation. We reviewed the effects of the most potent coronary vasodilatory drug groups, calcium antagonists and nitrocompounds, on coronary diameters. Intravenous or intracoronary injections of verapamil, diltiazem, nifedipine, nicardipine, and nisoldipine can cause profound coronary dilation which has been shown to be maximal with verapamil and nisoldipine. Shortcomings of calcium antagonists include short or unknown duration of action after bolus administration, severe drop in blood pressure, and lack of commercial availability of solutions for injection of many substances. Isosorbide dinitrate induces profound coronary dilation; however, after sublingual administration marked blood pressure decrease can occur, and the duration of action and ideal dose of intracoronary isosorbide dinitrate has not been investigated yet. Injections of molsidomine and its active metabolite, SIN-1, cause longlasting, reproducible, maximal coronary dilation, although only after a waiting period of at least 10 minutes; unfortunately, SIN-1 is only commercially available in France. Nitroglycerin induces reproducible maximal coronary dilation and is easy to administer sublingually or as intracoronary bolus injection with rapid onset of action and no major side effects. The short duration of action may require repeated administrations. To date, repeated intracoronary bolus injections of 0.1 mg nitroglycerin every 10 minutes seem to be the optimal known regimen for standardization of coronary vasomotor tone in serial angiographic studies. Further investigations in this field with old and new vasodilatory drugs are recommendable. PMID- 10453391 TI - Calculation of left ventricular outflow tract area using three-dimensional echocardiography. Influence on quantification of aortic valve stenosis. AB - In 23 patients with aortic valve stenosis (14 male, 9 female, mean age 66 +/- 21.5 years) left ventricular outflow tract cross-sectional area was determined in planimetric fashion using three-dimensional echocardiography. The 3-D data-set for each patient had been acquired in the course of a multiplane transesophageal examination. Aortic valve area was determined using the continuity equation. Results obtained were compared to those calculated by continuity equation using to the conventionally determined LVOT area (a = pi [d/2]2). As reference method the results were compared to invasive measurements. 3-D planimetric determination of LVOT cross-sectional area was possible in 20 of 23 patients. In three patients, this method failed due to artefacts. The mean difference to the conventionally calculated LVOT area amounted to 0.18 cm2 (SD = 0.46). The comparison of AVA determined by continuity equation and by invasive measurement showed a mean difference of -0.074 cm2 (SD = 0.21) for the conventionally calculated LVOT area; for the planimetrically determined LVOT area the mean difference of AVA amounted to -0.03 cm2 (SD = 0.14) (p < 0.05). Planimetric determination of LVOT area using 3-D echocardiography improves the agreement of the continuity equation with invasive measurement. PMID- 10453392 TI - Time-course of dobutamine-induced wall motion abnormalities in the infarct area following thrombolytic therapy. AB - Stress-induced asynergies in the infarct area following thrombolytic therapy are considered to reflect incomplete recanalization of the culprit vessel. However, reperfusion is a dynamic process with successive pathophysiological phases, so that the timing of assessment of residual ischemia may have relevant clinical implications. We studied the time-course of dobutamine-induced homozonal asynergies in 61 (group B) survivors of uncomplicated infarction as compared to 54 (group A) control subjects showing normal response to dobutamine stress echocardiography within 10 days of acute myocardial infarction. The 79 (43 of group A and 36 of group B) patients not presenting new cardiac events underwent further dobutamine stress echo within 90 +/- 17 days, which was positive in 20 and negative in 59. Persistence of test positivity was observed in just 17/36 (47%) patients, who showed significantly more extensive dobutamine-induced asynergies as compared to pre-discharge evaluation and less frequent (p < 0.01) evidence of viable myocardium. These results arise question about the decisional impact of stress-induced wall motion abnormalities in the culprit vessel area early after thrombolysis in low-risk patients and emphasize the need to further clarify the time factor role in this setting. PMID- 10453393 TI - Estimation of the right ventricular volume and ejection fraction by transthoracic three-dimensional echocardiography. A validation study using magnetic resonance imaging. AB - AIMS: To validate the use of three-dimensional transthoracic echocardiography compared with the magnetic resonance imaging for determination of right ventricular volume and ejection fraction. METHODS AND RESULTS: We recorded transthoracic echocardiographic images starting from the apical four-chamber view in which the RV is clearly visualized in 15 healthy volunteers. The scanning plane of the RV was obtained by the rotational scanning technique in 2 degree angular increments for three-dimensional reconstruction. The RV volumes in end diastole and end-systole were calculated using a Tomtec three-dimensional reconstruction computer. We also assessed the RV by cine magnetic resonance imaging using the Siemens Magnetom Impact Expert (1.0 T). Cine gradient echo images were obtained in the short axis of the RV. The RV volume at each phase was calculated by Simpson's method. We also calculated the RV ejection fraction. The RV volumes in end-diastole and end-systole were 111 +/- 22 ml and 52 +/- 13 ml, respectively as determined by three-dimensional echo, and 115 +/- 18 ml and 55 +/ 14 ml determined by MRI. The right ventricular volumes at end-diastole and end systole determined by three-dimensional echo were correlated with the volumes determined by MRI (r = 0.94 and 0.97, respectively, p < 0.001). The RV ejection fraction determined by three dimensional echo was also correlated with the ejection fraction determined by MRI (r = 0.90, p < 0.01). CONCLUSIONS: Three dimensional transthoracic echocardiography provided reliable calculations of the right ventricular volume and ejection fraction. PMID- 10453394 TI - Assessment of the normal adult right ventricular diastolic function using M-mode echocardiographic measurement of tricuspid ring motion. AB - Right ventricular function can be evaluated echocardiographically by assessing the longitudinal motion of the tricuspid ring recorded in the apical four chamber view. In this study, we applied this technique to assess the right ventricular diastolic function in 10 healthy Japanese men (mean age: 28 +/- 6 years; age range: 20-43 years). Echocardiographic studies were performed with a phased-array imaging system using a 2.5 MHz probe. Tricuspid ring motion was measured by two dimensional echo guided M-mode echocardiogram of the right lateral tricuspid ring. The excursion of the tricuspid ring during early diastole (DS; mm) and the peak rate of change of the excursion (dS/dt; mm/sec) were measured. We also assessed the right ventricular function by cine magnetic resonance imaging (MRI). Cine gradient echo images were obtained along the short axis of the right ventricle. The right ventricular volume at each phase of the cardiac cycle was calculated by Simpson's method and time-volume curves were constructed. The peak filling rate (dV/dt; ml/s) was determined from these time-volume curves. The dS was 12.8 +/- 2.5 mm, dS/dt was 132 +/- 27 mm/sec and dV/dt was 269 +/- 66 ml/s. There were significant positive correlations between dS and dV/dt (r = 0.80, P < 0.01), and between dS/dt and dV/dt (r = 0.45, P < 0.05). Based on our results, M mode echocardiographic measurements of tricuspid ring motion may be used to assess the right ventricular diastolic function. PMID- 10453395 TI - Intracardiac echocardiography: computerized detection of left ventricular borders. AB - A semi-automated method for two- and three-dimensional analysis of intracardiac echocardiography (ICE) images and image sequences is reported based on detection of epicardial and endocardial borders using graph searching. The border detection method was applied to 50 ICE images acquired in vivo in five dogs and to 108 images in 16 volumetric ICE image sequences from eight cadaveric pig hearts. The ICE images from the in vivo study showed good correlation between computer detected and observer-defined left ventricular (LV) cavity areas and epicardial areas (r = 0.99, y = 0.98x + 0.43 [cm2]; r = 0.99, y = 0.98x + 1.11 [cm2]; respectively). In the cadaveric hearts, the LV volumes were determined with the volume measurement error of 7.6 +/- 7.7% and 11.3 +/- 11.2% for the aortic valve and mitral valve image sequences, respectively. Our method facilitates an accurate and computationally efficient approach for the quantitative assessment of ICE image data in 2D and 3D. PMID- 10453396 TI - Comparison of radionuclide angiography with three echocardiographic parameters of left ventricular function in patients after myocardial infarction. AB - AIMS: Left ventricular function is an important outcome measure in patients with coronary artery disease, in particular in patients after myocardial infarction. It is reliably assessed by radionuclide angiography, but echocardiographic wall motion scoring might be an attractive alternative. METHODS: Four days after reperfusion therapy for acute myocardial infarction both radionuclide angiography and echocardiography were performed in 90 patients. Segmental wall motion scoring (WMSI) and visual estimation of the left ventricular ejection fraction (LVEF) was done by 2 independent observers. Repeated analysis was performed 1 month after the first reading. In 41 patients the LVEF was assessed quantitatively by tracing of endocardial outlines of the left ventricle. RESULTS: Both correlation with radionuclide angiography (estimated LVEF: r = 0.71, WMSI: r = -0.68, Tracing: r = 0.59) and inter- and intra-observer variability (estimated LVEF: 19% and 15%, WMSI: 65% and 59%) were in favour of the LVEF estimation method. Correlation with radionuclide angiography measurements was related to the quality of the echocardiogram and to the extent of coronary artery disease. CONCLUSION: Simple echocardiographic estimation of left ventricular ejection fraction in patients after reperfusion therapy for acute myocardial infarction proved to be superior to quantitative assessment of ejection fraction and to segmental wall motion scoring in comparison with radionuclide angiography. PMID- 10453397 TI - BMIPP-design and development. AB - In the early 1980s a major obstacle for myocardial SPECT using iodine-123-labeled fatty acids and imaging technology available at that time was the rapid metabolism and myocardial washout of activity. Development of the 15-(p iodophenyl)-3-(R,S)-methylpentadecanoic acid (BMIPP) fatty acid analogue was based on the established effects of methyl-branching in delineating the enzymatic aberration in Refum's disease and our early studies with the tellurium (Te) substituted fatty acid analogues. Extensive animal studies with the Te-fatty acids demonstrated that this major structural alteration did not affect initial myocardial extraction, but could successfully inhibit subsequent metabolism and significantly delay washout. Tracer kinetic evaluation and metabolic studies on experimental animals and Langendorff-perfused rat hearts clearly demonstrated that introduction of methyl-branching is an effective approach which alters tracer kinetics by delaying myocardial washout of radioiodinated fatty acids by increasing myocardial retention. Although irreversible retention of iodine-123 BMIPP is not observed, subsequent extensive human studies have clearly substantiated the delayed myocardial washout of BMIPP in comparison with the p IPPA straight chain analogue. Although contemporary SPECT capabilities allow much more rapid acquisition periods, the delayed washout is still a practical benefit in relation to the use of BMIPP. Most important, the unexpected mis-match which has been widely observed between perfusion tracer distribution and the regional BMIPP distribution (i.e. BMIPP < flow tracer) has been linked to the identification of jeopardized, but viable myocardial regions. In this paper the development of BMIPP is discussed and the results of recent studies focusing on evaluating the effects of the absolute configuration of the branched methyl group using the 3(R)-BMIPP and 3(S)-BMIPP are described. PMID- 10453398 TI - Basic kinetics of 15-(p-iodophenyl)-3-R,S-methylpentadecanoic acid (BMIPP) in canine myocardium. AB - BMIPP is a radioiodinated fatty acid analogue used for myocardial single photon emission CT (SPECT) imaging based on high cardiac fatty acid metabolism. In normal dogs, 74% of the injected BMIPP was instantly extracted and was then retained in 65.3%. The washout of the retained radioactivity was low, and most of the washout was alpha- and beta-oxidation metabolites. ATP concentration plays an important role in the myocardial uptake and retention of BMIPP. The ATP-dependent BMIPP uptake at the TG pool was strongly regulated by etomoxir with modifying mitochondrial beta-oxidation and subsequent ATP production. Thus, myocardial viability was reflected on the BMIPP uptake in acute ischemia. In spite of insignificant changes in early extraction and retention. BMIPP back diffusion (r = -0.92) and full-oxidation metabolite (r = 0.78) were correlated with the severity of ischemia. Mismatched region of BMIPP with flow (Tl-201) showed decreased metabolic enzymes such as citrate synthase and 3-hydroxyacyl-CoA dehydrogenase. These data suggest that BMIPP would be feasible for detecting cellular energy state from lipid metabolism. PMID- 10453399 TI - Cardiac BMIPP imaging in acute myocardial infarction. AB - Fatty acid metabolism functions as a major energy-producing system under aerobic conditions, but it is impaired immediately after myocardial ischaemia. This imaging can provide intracellular information which cannot be obtained by angiographical, perfusional or functional analysis. 123I-BMIPP and perfusion imagings in patients with acute myocardial infarction have demonstrated three different correlations between myocardial perfusion and fatty acid metabolism: concordant defects of perfusion and BMIPP which represent scar or non-viable tissue; lower BMIPP uptake relative to perfusion (perfusion-BMIPP mismatch) which implicates metabolically damaged, often dysynergic, but viable myocardium; and equivalently normal uptakes of perfusion and BMIPP in completely salvaged myocardium. Identification of these perfusion-metabolism correlations contributes to the detection of ischaemia-related myocardial injury in viable and non-viable myocardium, to the prediction of post-ischaemic or post-interventional functional recovery and to the identification of patients who have myocardium at ischaemic risk. Further clinical investigations might reveal more clearly the pathophysiological and prognostic implications of cardiac BMIPP imaging in patients with acute myocardial infarction. PMID- 10453400 TI - BMIPP imaging to assess functional outcome in patients with acute and chronic left ventricular dysfunction. AB - Assessment of myocardial viability is an important clinical issue for patient management during the acute and chronic stages of myocardial infarction. BMIPP (15-(p-iodophenyl)-3-(R,S)-methyl pentadecanoic acid) is a free fatty acid analogue which is trapped in the myocardium, thus permitting for metabolic imaging with single photon emission computerized tomography (SPECT). Less BMIPP than flow tracers that may be observed in the areas of infarction, may reflect the metabolic shift from fatty acid to glucose utilization in ischaemic myocardium. In this sense, the combined imaging of BMIPP and a flow tracer with SPECT may provide similar and important information as fluoro-18 deoxyglucose (FDG) and positron emission tomography (PET) regarding the assessment of myocardial viability. The purpose of this article is to review the clinical impact of BMIPP in patients with acute and with chronic left ventricular dysfunction for the identification of jeopardized but viable myocardium and the prediction of the functional outcome. PMID- 10453401 TI - BMIPP in angina pectoris. AB - Radionuclide imaging with BMIPP for assessment of myocardial metabolism can assist in clinical decision making in certain subsets of patients with coronary artery disease. In stable angina, abnormal BMIPP imaging and less BMIPP uptake than Tl-201 are more likely to be seen in the distribution of vessels with severe stenoses and related to regional wall abnormalities. BMIPP SPECT imaging is a promising approach in the evaluation of patients with suspected unstable angina during the pain-free period even after stabilization. Fatty acid metabolism is easily suppressed due to the abrupt onset of ischaemia. In addition, combined imaging with BMIPP and Tl-201 can provide insights into the understanding of abnormal metabolism in myocardium in stable and unstable angina. This review addressed the clinical applications of BMIPP imaging for evaluation of stable and unstable angina. PMID- 10453402 TI - beta-Methyl-p-(123I)-iodophenyl pentadecanoic acid single-photon emission computed tomography in cardiomyopathy. AB - beta-Methyl-p-(123I)-iodophenyl pentadecanoic acid (BMIPP) is one of the branched chain free fatty acids, which has suitable characteristics for myocardial SPECT because of higher uptake and longer retention in the myocardium. Recent advances of BMIPP myocardial SPECT for evaluating cardiomyopathy were reviewed. BMIPP defects were observed in 80% patients with hypertrophic cardiomyopathy (HCM). Moreover, BMIPP uptake was reduced at sites that corresponded with hypertrophic areas, where thallium uptake was increased. The correlations between severity score and septal wall thickness and LV function were better with BMIPP SPECT, suggesting that BMIPP is more suitable for the assessment of myocardial integrity in HCM. The dissociation between BMIPP and thallium defects was not observed frequently in dilated cardiomyopathy (DCM). We carried out BMIPP myocardial SPECT to evaluate the therapeutic effects of co-enzyme Q10 on DCM patients. Hearts to the mediastinum ratio and BMIPP defect scores were significantly decreased after co-enzyme Q10 treatment. BMIPP myocardial SPECT was confirmed to be sensitive in evaluating the therapeutic effect for the perspective of metabolic SPECT imaging. Recently, a lack of myocardial uptake of BMIPP has been found in a small subset of patients (0.3%-1.2%). Cardiac radionuclide imaging using BMIPP and 18F-FDG were performed on patients with type I CD36 deficiency. The percent dose uptake of 18F-FDG was significantly higher than in normal controls. CD functions as a major myocardial long-chain fatty acid transporter and its absence may lead to a compensatory upregulation of myocardial glucose uptake. An increased frequency of CD36 deficiency was demonstrated in cardiomyopathy. Therefore, fatty acid transport proteins and their related gene defects in relation to BMIPP uptake may become an important issue in the future. PMID- 10453403 TI - BMIPP compared with thallium redistribution. AB - I-123 labeled methyl-branched fatty acid, BMIPP, has been used mainly in Europe and Japan for the evaluation of various cardiac diseases. BMIPP uptake abnormality is usually more prominent tan perfusion, representing discordant BMIPP uptake less than thallium. This finding is observed in various cardiac disease conditions such as acute myocardial infarction, stable and unstable angina, hypertrophic and dilated cardiomyopathies. Many BMIPP studies combined stress thallium imaging in patients with coronary artery disease and hypertrophic cardiomyopathy demonstrated that such discordant BMIPP uptake less than thallium is related to stress-induced ischemia as evidenced by reversible thallium defect after exercise. BMIPP imaging is able to detect metabolic alteration in the heart which is not available by perfusion imaging alone. PMID- 10453404 TI - BMIPP compared with PET metabolism. AB - I-123 15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid (BMIPP) was introduced as an approved radiopharmaceutical in Japan in 1993. This article reviews our clinical comparisons between BMIPP and PET metabolism conducted at Kyoto University. BMIPP uptake was suggested to reflect exogenous fatty acid utilization in the myocardium. By the comparison between FDG PET and BMIPP SPECT, reduced uptake of BMIPP relative to thallium was considered metabolically damaged but viable myocardium in ischemic coronary artery disease. In hypertrophic cardiomyopathy, reduction of BMIPP uptake was shown to precede suppression of oxidative or glucose metabolism. Occasionally we encounter subjects with absent myocardial BMIPP uptake. The majority of these subjects showed metabolic switching from normal free fatty acid metabolism to abnormally enhanced glucose metabolism in the fasting state. Thus, BMIPP provides us with the valuable metabolic information which is unattainable using a perfusion tracer. PMID- 10453405 TI - Myocardial imaging with 123I-BMIPP in patients with congestive heart failure. AB - First, we studied the diagnostic utility of myocardial imaging with 123I-BMIPP (BMIPP), a 3-methyl-branched fatty acid analog, in patients with various types of cardiomyopathy and left ventricular dysfunction (ejection fraction below 40%) by comparing with myocardial flow tracer imaging. The incidence of a dissociation between myocardial BMIPP and 201Tl distributions (BMIPP < 201Tl) as a marker of metabolic abnormality in viable tissue varied considerably among various heart diseases. Patients with ischemic cardiomyopathy and the dilated form of hypertrophic cardiomyopathy had a higher incidence while those with idiopathic dilated, alcoholic and hypertensive cardiomyopathy had a lower incidence. These results suggest that the marked difference between ischemic and idiopathic dilated cardiomyopathies may contribute to the differential diagnosis between these two diseases which are main basic abnormalities in congestive heart failure. Second, we investigated the relationship between myocardial BMIPP uptake and ventricular stress in patients with right ventricular pressure overload due to pulmonary hypertension. Myocardial BMIPP uptake in the right ventricle estimated by referring to uptake in the left ventricle showed a significant correlation with mean pulmonary artery pressure (mPAP) and no significant difference with myocardial 99mTc-sestamibi uptake in the 15-81 mmHg mPAP range. These results suggest that myocardial utilization of free fatty acid may be preserved in the presence of higher ventricular wall stress. PMID- 10453406 TI - Future aspects of BMIPP. AB - Iodinated fatty acid compounds have an important role in early detection of myocardial abnormalities and provide insights into pathological states in the heart. Among them, 15-(p-iodophenyl)-3R,S-methyl pentadecanoic acid (BMIPP) has been most widely used providing excellent images of the left ventricular myocardium due to high myocardial uptake and long retention. The previous chapters have focused on the basic characters and clinical applications of this compound. However, the precise mechanisms of myocardial kinetics should be further investigated under various conditions. Most of the studies showed reduced BMIPP uptake relative to perfusion in a variety of myocardial disorders, whereas an increase in BMIPP uptake relative to perfusion is often reported. The potential mechanisms of such conflicting results are discussed, but basic studies should be performed to clarify such results in detail. There are a number of clinical values of this compound. Since alteration of fatty acid is observed in the repetitive ischemia, BMIPP can be used for detecting severe ischemic episodes. The concept of 'ischemic memory' imaging can be applied for patients with unstable or vasospastic angina at rest and for those with acute myocardial infarction with successful revascularization to identify the risk area. The discordant decrease in BMIPP uptake relative to perfusion is often seen in ischemic but viable myocardium, and therefore, the combined imaging of BMIPP and perfusion can be used for assessment of tissue viability. Furthermore, abnormal BMIPP uptake is most often observed in hypertrophic cardiomyopathy, and thus, this compound can be used for an early detection and differential diagnosis of the cardiomyopathy. Although BMIPP imaging seems to be quite promising in many fields, the number of patient data remain limited. In this respect, a multicenter study with a vast majority of patients is warranted to confirm these important values of BMIPP. In addition, this attractive tracer should be available all over the world to confirm its clinical value in the near future. PMID- 10453407 TI - Cardiovascular flow patterns: what should we make of them? PMID- 10453408 TI - Analysis of flow patterns using MRI. AB - This paper describes new software programs for analysis and visualization of blood flow patterns derived from time-resolved 3D velocity data sets. Using the programs, data can be displayed in cross-sectional or 3D perspective view. Particle paths revealing the flow patterns are computed by forward and backward time integration of the velocity field. Vector arrowmaps are computed as short duration paths starting from uniformly spaced points over the lumen volume. Background, divergence, and local boundary correction is done to improve the realism of the paths. The programs have been used to visualize flow patterns from non-gated and cardiac-gated 3D velocity enclosed data in over 35 subjects. Arrowmaps are preferred for revealing local regions of different blood flow characteristics within the vessel, while particle paths are preferred for revealing global organization of the flow. They are complementary display strategies. Advanced data handling and display features are essential for analyzing and visualizing large velocity encoded data sets. PMID- 10453409 TI - Complex flow patterns in the great vessels: a review. AB - The article reviews the applications of magnetic resonance velocity mapping based on phase shifts in the protons to quantify blood flow velocity and blood flow volume. The method can be used to study normal physiology of blood flow in the aorta and its major branches, including forward and backward flow, to measure the aortic valve function in aortic valvular disease, stenosis and regurgitation, as well as pulmonary artery flow velocities in pulmonic insufficiency and regurgitation. Superior vena cava flows, pulmonary vein flows, left-to-right shunts, atrial and ventricular pulmonary conduit flows can also be measured. Two- and three-directional velocity mapping is reviewed and can be used to study three or four-D flows in the aorta and the major arteries in great detail. PMID- 10453410 TI - Cardiovascular MR angiography. PMID- 10453411 TI - 3D contrast-enhanced MR angiography using fluoroscopic triggering and an elliptical centric view order. AB - Although a variety of timing techniques and data acquisition strategies have been used for three-dimensional contrast-enhanced MR angiography, many are still limited by inadequate overall reliability, limited spatial resolution, or complexity. A technique is presented in this work in which contrast arrival is detected in the targetted vasculature in real time using MR fluoroscopy. Upon detection the operator triggers a 3D MR angiographic acquisition which uses an elliptical centric view order. It is shown that the view order intrinsically provides a high degree of venous suppression which in turn allows acquisition times of 30 seconds or longer, permitting high spatial resolution. The reliability of fluoroscopic triggering in bolus detection is shown to be over 90%. The technique provides high quality contrast-enhanced MR angiograms for many vascular regions. PMID- 10453412 TI - Contrast enhanced body magnetic resonance angiography. AB - The use of gadolinium contrast for body MRA is reviewed. Considerations for timing of the bolus of contrast are discussed. The utility of this technique is illustrated through clinical examples. Contrast enhanced MRA is rapidly replacing conventional angiography for many applications. PMID- 10453413 TI - Breathhold cardiac MRI and MRA. AB - Recent developments in MRI software and hardware have increased the speed and versatility of cardiac MRI by allowing image acquisition in a single breathhold. Many studies have shown that conventional cardiac MRI is as accurate as echocardiography or cardiac catheterization for diagnosing numerous cardiac conditions. In many cases cardiac MRI is the most accurate diagnostic technique but has not been widely adopted for routine cardiac imaging. One reason why the use of cardiac MRI has been limited is the long examination times required for conventional cardiac MRI. The development of better hardware, such as faster gradient amplifiers and dedicated surface coils, has allowed the implementation of much faster EKG-gated imaging sequences. These can be used in a single breathhold period, with a significant improvement in image quality compared with conventional sequences. Breathhold sequences can provide all the information provided by conventional cardiac MRI in a shorter time and with equal or better accuracy. Breathhold imaging will allow much wider application of MRI to routine cardiac diagnosis. PMID- 10453414 TI - Magnetic resonance imaging of complex congenital heart disease. AB - Current MR techniques enable both anatomical and functional evaluations of the heart. Although it is rarely used as a primary diagnostic tool in pediatric cardiology, it can be used as a valuable adjunct to echocardiography and angiography. MRI is particularly useful in clarification of morphology of complex congenital heart diseases. It is the most accurate method of determination of visceral and atrial situs. It is easy to evaluate the systemic and pulmonary venous connections that are very important for the Fontan type of operation, especially in patients with visceral heterotaxy. It facilitates demonstration of diminutive pulmonary arteries in patients with pulmonary atresia. It clearly demonstrates juxtaposition of the atrial appendages that is often missed by echocardiography. The anatomy of the twisted atrioventricular connections is much more clear in MRI than in echocardiography. It enables en face imaging of ventricular septal defect that provides a surgical view. We find en face imaging particularly helpful in patients with double inlet left ventricle and transposition of the great arteries where the size of the ventricular septal defect governs the blood flow into the aorta. It is often advantageous to echocardiography in defining the type of univentricular atrioventricular connections by demonstrating the position and topology of the rudimentary chamber in difficult cases. In double outlet right ventricle, the spatial relationship of the ventricular septal defect to the great arterial valves can be clearly defined by visualizing the ventricular septal defect and the outlet septum in the same imaging plane. PMID- 10453415 TI - Gadolinium contrast-enhanced three-dimensional MRA of peripheral arteries with multiple bolus injection: scan optimization in vitro and in vivo. AB - In this study, a scanning protocol was developed to image the arterial bed of the pelvis and both legs along their entire length in patients with peripheral arterial disease, using standard hard- and software. Three adjacent stations are acquired consecutively, with some small overlap; per station; one Gadolinium contrast bolus is administered. The scanning protocol was optimized in an in vitro phantom study. The optimal flip angle was found to be 50 degrees. Also, the optimal scan delay was chosen to be equal to the arrival time of the contrast bolus thereby minimizing artifacts. Three contrast bolus injections showed sufficient enhancement of the vessels after image subtraction. Finally, stenosis quantification by manual caliper was performed by five observers in the MRA images and correlated with the percent diameter reduction determined by quantitative angiography from corresponding X-ray images. The results of the MRA measurements were reproducible and intra- and inter-observer variabilities were statistically non-significant (p = 0.54 and p = 0.12, respectively). Stenosis quantification performed by four observers showed a good correlation with the X ray derived values (rp > 0.90, p < 0.02); the results from one observer were not significantly correlated. Five patients with proven peripheral disease were investigated with this new MRA scanning protocol. The images were of good quality which allowed adequate clinical evaluation; the original diagnoses obtained from X-ray examinations, were confirmed with MRA. In conclusion, peripheral arterial disease can be evaluated adequately with this MR scanning protocol. PMID- 10453416 TI - New black blood pulse sequence for studies of the heart. AB - The black blood concept is based on the signal void principle of the sequences in Spin Echo, SE or FSE, which are very useful for studying the mediastinum and heart. In this setting, new sequences are continuously introduced to eliminate the artifacts caused by breathing and heart movements. One such sequence is the Double-IR preparation Black Blood FSE. We report our experience in 97 patients, using this new pulse sequence to evaluate cardiac pathology, and establish comparisons with the conventional Spin Echo sequences. The study comprises mediastinal disease and aorta and heart explorations. We consider this new Double IR preparation FSE sequence to be an excellent choice for evaluating chest, mediastinal and cardiac images. The sequence offers improved spatial resolution of both the vessels and other chest structures with respect to conventional Spin Echo imaging. With the exception of patients presenting severe heart problems, or in the presence of intense bradycardia, the required 16 cycles in apnea are well tolerated. The purpose of the present study is to present our initial results with this new pulse sequence as applied to cardiac pathology, in comparison with conventional Spin Echo imaging. PMID- 10453417 TI - Development of a transgenic mouse model using rat insulin promoter to drive the expression of CRE recombinase in a tissue-specific manner. AB - BACKGROUND: Tissue-specific ablation of a gene using the Cre-loxP system has been used as an important tool to define its role, in addition to the total ablation, to avoid the embryonic lethality in case of wide expression of the target gene. METHODS: The RIP-Cre genetic construct was generated by standard subcloning techniques and microinjected into one cell embryo to develop the transgenic mouse line. Transgenic mice were screened by polymerase chain reaction (PCR) using DNA isolated from tell digestion. Tissue specificity of RIP was demonstrated by transient transfection of RIP-1acZ construct to NIT-1 cells (mouse insulinoma cell line) in vitro. RESULTS: The 448 nucleotides of RIP were sufficient for beta cell specific expression of the reporter gene as evidenced by the presence of blue color in the nucleus of NIT-1 cells. Isolated RIP-Cre transgene was microinjected, and PCR screening identified two independent lines of transgenic mice. Tissue specificity of RIP was demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR) using the islet RNA from the transgenic mice. CONCLUSION: We have established a tissue-specific transgenic mouse model using Cre recombinase linked to rat insulin promoter (RIP) to drive the expression of the reporter gene specifically in the beta-cells. The RIP-Cre transgenic mice will allow beta-cell specific ablation of target gene(s) to define its role in the regulation of islet physiology. PMID- 10453418 TI - Different patterns in immunoreactive anionic and cationic trypsinogen in urine and serum in human acute pancreatitis. AB - BACKGROUND: Acute pancreatitis (AP) results in elevated concentrations of trypsinogen (T) isoenzymes in serum. Immunoreactive anionic trypsinogen in urin (irAT/u) is elevated in AP, and has recently been proposed as a rapid diagnostic instrument and severity predictor. These results have not been confirmed by other groups, and irAT/u has not been further characterized. The concentration of immunoreactive cationic trypsinogen in urine (irCT/u) and the serum irAT/irCT ratio in AP have not been extensively examined. METHODS: Levels of irAT and irCT were studied in urine and serum from 50 AP patients and in urine from 41 non-AP patients. Severity was assessed according to the Atlanta classification. irAT/u was characterized by gel filtration. RESULTS: Gel filtration revealed only AT in the urine. Highly significant differences in irAT/u were seen between AP/non-AP (p < 0.0001) and mild/severe disease (p = 0.0012). The irAT/irCT ratio in serum changed from normal 0.8 to 1.3 in AP. CONCLUSIONS: IrAT and only traces of irCT were found in the urine in AP. IrAT/u was higher in AP than in other acute abdominal disorders (non-AP) and also higher in severe than in mild AP. IrAT in serum (irAT/s) increased proportionally more than irCT/s in AP, but did not discriminate mild from severe forms. High levels of irAT/u in some non-AP cases and a wide range in AP cases make the clinical value of the test questionable. PMID- 10453419 TI - Treatment of pancreatic exocrine insufficiency after pancreatic resection. Results of a randomized, double-blind, placebo-controlled, crossover study of high vs standard dose pancreatin. AB - BACKGROUND: Steatorrhea following major pancreatic resection can be difficult to control, requiring high doses of pancreatic enzyme supplements. The aim of this study was to demonstrate equivalent efficacy of high-dose and standard-dose pancreatin in treating steatorrhea after pancreatectomy. METHODS: A randomized, double-blind, crossover study was conducted with a 2-wk run-in period for stabilization on a suitable dose of standard-dose pancreatin and two 14-d treatment periods using either high-dose or standard-dose pancreatin at this dosage. Parameters used to demonstrate efficacy of treatment were stool fat excretion, stool volume, and clinical symptoms. RESULTS: Thirty-nine patients who had undergone total or partial pancreatectomy were randomised; 37 completed all parts of the study. During stabilization, the mean daily capsule intake was 19.4 (range 9-54); even so, 22 (56%) patients had stool fat excretion > 7 g/d. There were significant correlations between stool fat excretion and stool volume (p < 0.0001) and stool frequency (p < 0.01), but not with indices of abdominal pain and global symptoms. Both high-dose and standard-dose pancreatin demonstrated statistically similar efficacy in the treatment period. CONCLUSION: The use of high-dose pancreatin for the treatment of pancreatic insufficiency in patients following pancreatectomy should significantly reduce capsule intake with increased convenience and greater compliance rate. Our results, however, indicate that further progress is needed to resolve steatorrhea following pancreatic resection. PMID- 10453420 TI - One thousand faces of Langerhans islets. AB - Studies with different strains of Syrian hamsters and Syrian golden hamsters have revealed the remarkable potential of islet cells to undergo orthotopic and heterotopic metaplasia. The most common spontaneous change included the development of hepatocytes in aged and malnourished hamsters. Of the many other alterations that occurred during carcinogenesis, most of the metaplastic changes originated within the islet periphery and progressed inside and outside the islets. The development of ductular structures within islets and their progression either to structures identical to human serous cystadenoma or to highly invasive adenocarcinomas were the most common alterations. The remarkably greater invasive potential of cancer cells arising within the islets contrasted sharply with the slow growth of the tumors developing within ducts (intraductal tumors). Studies in human tissue also showed development of malignant cells within islets, and, in some cases, transition of islet cells to malignant cells was suggested. The overall results, along with recent findings in other studies in cultured human and hamster islets, indicate the enormous potential of islet cells to differentiate and undergo malignant transformation. Whether the metaplastic and malignant cells derive from stem cells embedded within islets or from transdifferentiated islet cells remains to be seen. PMID- 10453421 TI - Diagnosis, objective assessment of severity, and management of acute pancreatitis. Santorini consensus conference. AB - BACKGROUND: The diagnosis, early assessment, and management of severe acute pancreatitis remain difficult clinical problems. This article presents the consensus obtained at a meeting convened to consider the evidence in these areas. The aim of the article is to provide outcome statements to guide clinical practice, with an assessment of the supporting evidence for each statement. METHOD: Working groups considered the published evidence in the areas of diagnosis, assessment of severity, nonoperative treatment, and surgical treatment of severe acute pancreatitis. Outcome statements were defined to summarize the conclusions on each point considered. The findings were discussed and agreed on by all participants. A careful assessment was made of the strength of the available evidence (proven, probable, possible, unproven, or inappropriate). FINDINGS AND CONCLUSIONS: There is reliable evidence to support much current practice. Clear guidance can be given in most areas examined, and several areas were identified where further investigation would be helpful. Diagnosis using plasma concentrations of pancreatic enzymes is reliable. Rapid advances are taking place in the assessment of severity. Several new therapeutic strategies show real promise for the reduction of morbidity and mortality rates. Surgical debridement is required for infected pancreatic necrosis, but is less often necessary for sterile necrosis. PMID- 10453422 TI - Proximal migration of transanastomotic pancreatic stent following pancreaticoduodenectomy and pancreaticojejunostomy. AB - BACKGROUND: The use of catheters to stent the pancreaticojejunal anastomosis following pancreaticoduodenectomy is practiced by some surgeons. Their long-term effects in this setting, however, remain unknown. METHODS: A 60-yr-old woman underwent a potentially curative pylorus preserving pancreaticoduodenectomy for Stage I ampullary carcinoma. Roux-en-Y pancreaticojejunostomy was constructed over a short stent. She presented 4 yr later with abdominal pain, steatorrhea, and weight loss. Computed tomography revealed a stent within the proximal pancreatic duct, with gross upstream ductal dilatation and parenchymal features of chronic pancreatitis. RESULTS: Laparotomy revealed no disease recurrence. The stent, removed through a jejunotomy, was occluded. On-table pancreatogram demonstrated a 3-cm proximal duct stricture. Drainage was achieved with a lateral pancreaticojejunostomy (modified Puestow procedure). Recovery was uneventful, with clinical recovery of pancreatic exocrine function at 6 mo follow-up. CONCLUSION: Proximal migration of transanastomotic pancreatic stent with subsequent development of chronic pancreatitis is a potential complication following pancreaticoduodenectomy. It can be managed effectively with stent removal and a lateral pancreaticojejunostomy. PMID- 10453423 TI - Acute pancreatitis secondary to 5-aminosalicylic acid therapy in a patient with ulcerative colitis. AB - Therapy with oral 5-aminosalicylic acid (5-ASA) for ulcerative colitis has been reported to be effective and safe. We describe a case of biochemically proven mild acute pancreatitis occurring after 9 d of oral 5-ASA therapy for ulcerative colitis. A hypersensitivity mechanism seemed to be involved in the development of pancreatitis probably owing to erratic systemic absorption of the drug. We suggest clinical and biochemical monitoring for early diagnosis of pancreatitis in patients with ulcerative colitis receiving 5-ASA administration. This is the first report of acute pancreatitis developed by oral 5-ASA therapy for the treatment of ulcerative colitis in the literature of Japan. PMID- 10453424 TI - Lymphoepithelial cyst of the pancreas. No evidence for Epstein-Barr virus-related pathogenesis. AB - Compared to pseudocyst formation after prior pancreatitis, true cysts of the pancreas are rare. Pancreatic cysts with irregular wall components or a mucinous content raise the suspicion for the presence of a cystic neoplasm, and surgical resection is recommended. A case of a patient with a history of prostate cancer is described in whom a cyst of the pancreatic tail was discovered incidentally. Based on the radiographic features, which did not support the presence of a serous cystadenoma, a spleen-preserving distal pancreatectomy was performed. Histologic features were characteristic for a lymphoepithelial cyst (LEC) of the pancreas, lined with thinned squamous epithelium surrounded by benign lymphoid tissue. Since LECs of the parotid gland, which are associated with acquired human immunodeficiency, are frequently related to Epstein-Barr virus (EBV) infection, EBV in situ hybridization was performed and did not reveal evidence for EBV. Twenty-eight instances of pancreatic LECs have been reported, primarily affecting adult males, without evidence of increased numbers of EBV-positive cells. The pathogenesis, differential diagnosis, and clinical implications of lymphoepithelial pancreatic cysts are discussed. PMID- 10453425 TI - The "two-hit" pathogenetic concept of chronic pancreatitis. PMID- 10453426 TI - New concepts in understanding the pathophysiology of chronic pancreatitis. PMID- 10453427 TI - Combined high-intensity strength and aerobic training in diverse phase II cardiac rehabilitation patients. AB - BACKGROUND: Developing adequate levels of muscular strength in the cardiac rehabilitation (CR) patient helps return the patient to an active lifestyle. This study evaluated the effects and safety of an 8-week high-intensity strength training (ST) program combined with a traditional aerobic-based CR program on the muscular strength of a diverse phase II CR population. METHODS: Sixty-one phase II CR patients (age = 60.5 +/- 10.6 years) stratified by risk (high risk: n = 18, ejection fraction = 23.6 +/- 7.8%; intermediate risk: n = 19, ejection fraction = 40.0 +/- 4.6%; low risk: n = 24, ejection fraction = 58.0 +/- 7.7%) and gender (males = 46, females = 15) participated. One repetition maximum (1RM) testing was performed on the horizontal squat, shoulder press, leg extension, lat pulldown, and biceps curl. Patients performed two sets of each exercise 2 days per week at an intensity that started at 60% 1RM and progressed to 80% 1RM by week 4. Weeks 4 to 8 intensity was adjusted individually to maintain 8RM per set. Blood pressure and heart rate/rhythm responses to 1RM testing were monitored in high-risk patients. Muscle soreness and injury were monitored for all patients immediately after 1RM testing and on days 2 and 7. RESULTS: All patient groups made significant gains (P < 0.05) in muscle strength (mean increase: lower body = 15.3%, upper body = 16.7%). No injury or significant muscle soreness occurred due to 1RM testing. No abnormal heart rate/rhythm or blood pressure responses occurred in high-risk patients. CONCLUSIONS: Diverse phase II CR patients can improve their strength significantly with a combination of high-intensity strength and aerobic training. PMID- 10453428 TI - Long-term benefits of exercise maintenance after outpatient rehabilitation program in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Optimal strategies to maintain short-term benefits of an initial rehabilitation program (RP) are not known. To assess the long-term effects of exercise maintenance (EM) after an initial outpatient RP, the authors conducted a prospective study. PATIENTS AND METHODS: Fifty-eight patients with moderate to moderately severe chronic obstructive pulmonary disease who completed an initial 7-week outpatient RP were included. They were allocated into four groups according to the conditions of EM they self-selected: 15 patients received a structured EM session supervised by a physiotherapist twice a week (group A); 14 patients received a structured EM session supervised by a physiotherapist once a week (group B); 15 patients continued self EM daily at home (group C); and 14 patients did not continue EM (group D). Patients were evaluated before, immediately after, and 18 months after the initial outpatient RP. Measurements included exercise testing on a cycle ergometer and a visual analog scale to evaluate chronic dyspnea. RESULTS: After RP, all patients exhibited improvements in maximal workload (P < 0.05) and in dyspnea (P < 0.05). Improvements in maximal workload were maintained at 18 months in patients in groups A, B, and C but these only reached significance in groups B and C (P < 0.05). On the other hand, maximal workload returned to baseline values in group D (P = 0.01) at 18 months. CONCLUSION: Our results indicate definite benefits of EM after an initial outpatient RP compared with no EM. Daily EM at home appears to be as efficient as structured EM sessions supervised by a physiotherapist, once or twice a week, in moderate to moderately severe chronic obstructive pulmonary disease. PMID- 10453429 TI - Lifestyle modification program in management of patients with coronary artery disease: the clinical experience in a tertiary care hospital. AB - OBJECTIVES: The authors examined clinical outcomes in 71 male and female patients with coronary atherosclerosis who enrolled in a 2-year, independent-living, lifestyle modification program. The findings in 43 patients who completed the program were compared with those in 28 patients who dropped out of the program. BACKGROUND: Clinical studies suggest that lifestyle modification of risk factors for coronary atherosclerosis reduces subsequent cardiac events but there are very few reports of the effect of these programs in patients living independently. METHODS: Patients with diagnosed coronary atherosclerosis were managed for a 2 year period in a structured multidisciplinary program by a team that included two cardiologists, a nurse, a dietitian, an exercise physiologist, and a clinical psychologist. The overall aim of the program was to normalize or control all major reversible cardiovascular risk factors. Patients were required to participate in several weekly sessions for exercise, meditation/stress reduction training, dietary education and counseling, and participatory dinners. There was a strong emphasis on patient's self care, inclusion of support members, and regular monitoring of and feedback to patients. RESULTS: Data comparing baseline and 2-year outcomes showed a significant reduction in body weight, dietary intake of total/saturated fat and cholesterol, serum low- and high-density lipoprotein concentration, and an increase in exercise capacity. In the compliant group, the incidence of cardiac events was 2.3% over 2 years. CONCLUSION: Multidisciplinary lifestyle modification programs addressing cardiovascular risk factors are known to have a significant impact upon cardiac risk factors in patients with coronary atherosclerosis. Data show that these changes can be accomplished in independent living patients in a program offered through a routine cardiology service. However, compliance is an important issue in these self-regulated programs. PMID- 10453430 TI - Feasibility and efficacy of home exercise training before lung volume reduction. AB - BACKGROUND: Exercise training is recommended before lung volume reduction surgery (LVRS) in patients with emphysema. Unfortunately, many of these patients are referred from remote areas where there is no available rehabilitation program. The authors evaluated the feasibility and efficacy of a minimally supervised home based exercise training program. METHODS: Twenty-three emphysematous patients (age 61 +/- 6, forced expiratory volume in 1 second = 29 +/- 7% predicted [mean +/- SD]) were recruited from our LVRS program. Measurements of pulmonary function, maximal and submaximal exercise capacity, 6-minute walking distance (6 MWD), muscle strength, and quality of life with the Chronic Respiratory Questionnaire were obtained before and after training. Home-based exercise training program included muscle exercises and aerobic training, and started with detailed teaching while the follow-up was ensured through weekly phone calls and a diary filled by each patient. RESULTS: Significant increases in 6-MWD (P < 0.001), quality of life (P < 0.005), peak work rate (P < 0.05), peak oxygen consumption (P < 0.05), endurance time (P < 0.005), and muscle strength were observed in the home-based exercise training program. CONCLUSIONS: Home-based exercise training for patients in preparation for LVRS was feasible, and induced significant improvement in exercise tolerance and quality of life. PMID- 10453431 TI - Accuracy of recall of dyspnea after exercise training sessions. AB - BACKGROUND: Although clinicians often rely on patients' retrospective reporting of dyspnea, it is not known if dyspnea scores recalled after exercise are equivalent to dyspnea scores during exercise. The objective of this study was to determine whether patients could accurately recall after exercise the maximum ratings of the intensity of dyspnea and the anxiety associated with it that they experienced during exercise. METHODS: Forty-nine patients with chronic obstructive pulmonary disease (COPD) (forced expiratory volume in 1 second 0.92 +/- 0.23 L) participating in a randomized clinical trial of the impact of coached versus monitored exercise training on dyspnea rated dyspnea intensity (shortness of breath [SOB]) and dyspnea-related anxiety (DA) on a visual analog scale every 2 minutes during treadmill exercise. After each of 12 training sessions each subject was asked to rate the worst level of the two sensations that they recalled having experienced during exercise. RESULTS: For the groups as a whole, actual maximum scores for SOB and DA during exercise were highly correlated with recalled maximum values after exercise (r > or = 0.85, P < 0.0001) and the average differences were small (0-10.9 mm on a 200-mm scale). However, individual variation was substantial, limiting predictability for individual ratings. CONCLUSIONS: After exercise, patients with COPD as a group can accurately recall the worst SOB and DA that they experienced during exercise. This finding supports the further study and use of retrospective symptom ratings as a method for dyspnea assessment during exercise training in pulmonary rehabilitation. PMID- 10453432 TI - Non-physician supervision of cardiopulmonary exercise testing in chronic heart failure: safety and results of a preliminary investigation. AB - PURPOSE: In the current era of efficient use of personnel and cost containment, the use of non-physicians in selected roles previously occupied exclusively by physicians has become increasingly prevalent. Traditionally, physicians have directly supervised graded exercise testing of patients with chronic heart failure. The purpose of this prospective pilot investigation was to determine the safety and results of non-physician supervised exercise testing of these high risk patients. METHODS: Two hundred eighty-nine consecutive outpatients (211 men, 78 women) with left ventricular ejection fractions of < or = 35% were referred for cardiopulmonary exercise testing. Symptom-limited treadmill graded exercise tests were supervised by paramedical personnel with a physician immediately available, but not present in the lab. RESULTS: Nonsustained ventricular tachycardia was present during exercise in approximately 20% of patients. Test limiting hypotension was documented in 5% of subjects. Only one serious event occurred during the 289 exercise tests, an episode of ventricular fibrillation with a successful resuscitation outcome. Peak exercise respiratory exchange ratio averaged 1.10 +/- 0.14, consistent with a near-maximal patient effort. Peak oxygen uptake was 18 +/- 5 ml/kg/min. CONCLUSIONS: Supervision of cardiopulmonary graded exercise testing in properly screened patients with severe systolic left ventricular dysfunction by experienced non-physicians appears to be reasonably safe and the results are suitable for clinical decision making. Such a practice is an attractive cost-containment strategy and deserves further investigation. PMID- 10453433 TI - Lipid lowering in the cardiac rehabilitation setting. AB - PURPOSE: The authors determined the frequency and effectiveness of pharmacologic lipid lowering, guided by the recommendations of the National Cholesterol Education Program (NCEP) before and after institution of a systematic lipid assessment performed at the time of the cardiac rehabilitation entry evaluation. METHODS: The systematic lipid evaluation included a full lipid profile and a dietary evaluation at which time an active approach to pharmacologic lipid therapy was taken. Therapy was guided by the NCEP guidelines, with the collaboration of the referring physician. The frequency of lipid therapy change (starting or changing therapy) from the baseline evaluation to a 3-month follow up visit was the primary study outcome variable. The control group consisted of 51 patients with coronary heart disease (CHD) seen in 1995 at cardiac rehabilitation, who agreed to have their serum lipids measured in a double blinded fashion. There was no systematic lipid lowering intervention. The intervention group consisted of 187 patients with CHD who participated in cardiac rehabilitation in 1996 to 1997. RESULTS: At baseline, a similar percentage of patients in each group were on lipid lowering therapy: 38% (19/51) in controls versus 35% (65/187) in intervention patients. Among patients with a baseline low density lipoprotein (LDL) cholesterol of > or = 130 mg/dL, therapy was modified in 18% (4/22) of control patients compared with 52% (35/68) of intervention patients (P < 0.05). Among patients with a baseline LDL cholesterol of > or = 160, therapy was altered in 22% (2/9) control patients compared with 72% (18/25) intervention patients (P < 0.01). In both risk strata of > or = 130 mg/dL and > or = 160 mg/dL, LDL cholesterol measures were lowered to a greater degree in the intervention group. CONCLUSIONS: The performance of a systematic lipid review at the time of cardiac rehabilitation entry, with an active stance toward pharmacologic therapy, results in a threefold increase in pharmacologic modifications and lower LDL cholesterol values for cardiac rehabilitation participants. PMID- 10453434 TI - Risk stratification for exercise training in cardiac patients: do the proposed guidelines work? PMID- 10453435 TI - Nurture with ionizing radiation: a provocative hypothesis. AB - Whole body exposure to low-dose ionizing radiation appears to decrease overall cancer incidence. The data come from at least eight large studies of populations exposed to various forms of radioactive material and from more limited studies of occupational and environmental exposures to plutonium, radium, and radon. Earlier experiments in animals strongly support the protective effect that is apparent in humans. Also, experimental reports from invertebrates kept in radiation-deficient conditions suggest that ionizing radiation is essential for optimal growth and development. The combined evidence points to the presence of no-adverse-effect thresholds and of hormesis or beneficial effects at doses below those thresholds. Furthermore, according to the geological record, the high background radiation under which life first evolved has progressively attenuated up to the present. Thus it is intriguing to postulate that modern humans may live under conditions of partial deficiency of ionizing radiation; low doses of ionizing radiation may likely function as inducers of repair and detoxification mechanisms, much as low level antigenic challenges are responsible for enhanced immune competence. This hypothesis runs contrary to the prevailing consensus of regulatory default assumptions, which negate the possibility of no-effect thresholds for agents that are carcinogenic at certain levels of exposure. Still, those are dogmatic policy assumptions without scientific or even empirical justification, whereas the hypothesis advanced here has consistent observational and experimental support. The implication is that a partial deficiency of ionizing radiation could be remedied by a safe supplementation, possibly through dietary means. Dose-response data from studies of nuclear workers and populations subjected to unusual exposures suggest that safe supplementation with about 0.4 cGy/mo would be beneficial and conservative. PMID- 10453436 TI - Genistein-induced cell cycle arrest and apoptosis in a head and neck squamous cell carcinoma cell line. AB - Epidemiological studies have shown a lower incidence of breast, prostate, and colon cancers in Asian countries, particularly China and Japan, than in the United States. It is believed that genistein, a natural tyrosine kinase inhibitor and a metabolite of soy products, may be responsible for the protection from these cancers. Genistein was shown to inhibit cell proliferation and to induce cell cycle arrest at the G2-M phase in breast, prostate, and jurkat T cell leukemia cell lines. However, such studies have not been reported in squamous cell cancers of the head and neck. In this report, we show that genistein inhibits proliferation of a squamous cell carcinoma cell line HN4. Additionally, genistein caused cell cycle arrest at the S/G2-M phase and induced programmed cell death (apoptosis) in these cells. These effects appear to be dose and time dependent, irreversible, persisting when the cells were recultured in genistein free medium for up to 72 hours, and specific for tumor cells, because genistein did not affect normal keratinocytes. These results suggest that if genistein shows similar results in clinical trials, it can be a potential chemopreventive/chemotherapeutic agent for cancers of the head and neck. PMID- 10453437 TI - A case-control study of diet and testicular carcinoma. AB - No risk factor other than cryptorchidism has been consistently associated with testicular cancer, and the influence of diet on testicular cancer risk has not been extensively explored. A few studies have found increased testicular cancer risk in men whose diets are high in fat, red meats, and milk or low in fruits and vegetables. We evaluated the relationship of dietary factors and risk of testicular cancer and also examined whether this risk varied by type of testicular cancer. We conducted a hospital-based case-control study at The University of Texas M. D. Anderson Cancer Center (Houston, TX) of 160 testicular cancer cases diagnosed between 1990 and 1996 and 136 friend-matched controls. The results of multivariable logistic regression analysis showed that after adjustment for age, education, income, ethnicity, cryptorchidism, and total daily calories, increasing total fat, saturated fat, and cholesterol consumption were associated with increasing risk of nonseminoma testicular cancer, with odds ratios (ORs) for the highest vs. the lowest quartiles of 6.3, 5.3, and 4.6, respectively. The risk for seminoma testicular cancer marginally increased with increasing intake of total fat and saturated fat, with ORs for the highest vs. lowest quartiles of 1.9 and 2.1, respectively. Higher total fat consumption was nearly significantly related to increased mixed germ cell tumor risk, with an OR for highest vs. lowest quartile of 4.2. This study supports the hypothesis that diet (particularly high fat consumption) increases testicular cancer risk in young men. However, the small sample size and the possibility that these observations may be due to bias indicate that the relationship of diet and testicular cancer risk needs to be further examined within a prospective or incident case-control study. PMID- 10453438 TI - A clinical trial to selectively change dietary fat and/or energy intake in women: the Women's Diet Study. AB - Dietary fat and energy intake have been implicated in breast cancer etiology. To examine the relative importance of these dietary factors on markers of cancer risk in women, we designed an intervention trial to selectively decrease fat and/or energy intake in free-living, premenopausal women who were somewhat overweight. The study used a 2 x 2 factorial design to evaluate the independent and interactive effects of dietary fat and energy. The diets were nonintervention, low fat (15% of energy from fat, maintenance of energy intake), low energy (25% energy reduction), and combination low fat and low energy. We utilized an individualized counseling approach with self-selection of foods. Women on the low-fat and combination diets were asked to meet given daily goals for fat grams and food group exchanges, while women on the low-energy diet used only food group exchanges. Of the 113 premenopausal women randomized who were eligible for analysis, 43% were African-American. A total of 88 women completed the 12-week program, and adherence to the dietary goals was similar in both racial groups. Women on the low-fat diet were able to reduce dietary fat intake to 19% of energy by 4 weeks and to 17% by 12 weeks with a slight decrease in energy intake. Women on the low-energy diet met their energy reduction goals by four weeks while maintaining percentage of energy from fat. Women on the combination diet largely met their goals by four weeks as well. These data indicate that it is possible to selectively manipulate dietary fat and energy intake in women over a short period of time, which makes clinical studies on the relative effects of these two dietary variables on cancer risk biomarkers readily feasible. PMID- 10453439 TI - Oral niacin prevents photocarcinogenesis and photoimmunosuppression in mice. AB - Topical nicotinamide (niacinamide) has demonstrable preventive activity against photocarcinogenesis in mice. To better understand how this vitamin prevents ultraviolet (UV) carcinogenesis, we tested systemic administration of another form of the vitamin, niacin, and its capacity to elevate cutaneous nicotinamide adenine dinucleotide (NAD) content as well as to decrease photoimmunosuppression and photocarcinogenesis. BALB/cAnNTacfBR mice were fed the AIN-76A diet supplemented with 0%, 0.1%, 0.5%, or 1.0% niacin throughout the experiment. UV irradiation consisted of five 30-minute exposures per week to banks of six FS40 Westinghouse sunlamps for 22 weeks in the carcinogenesis experiments, yielding a total cumulative dose of approximately 1.41 x 10(6) Jm-2 of UV-B radiation. Dietary supplementation with 0.1%, 0.5%, or 1.0% niacin reduced the control incidence of skin cancer from 68% to 60%, 48%, and 28%, respectively, at 26.5 weeks after the first UV treatment. Two potential mechanisms by which niacin prevents tumor formation were identified. Photoimmunosuppression, critical for photocarcinogenesis, is measured by a passive transfer assay. Syngeneic, antigenic tumor challenges grew to an average of 91.6 +/- 19.7, 79.8 +/- 11.5, 41.9 +/- 11.7, or 13.2 +/- 4.1 mm2 in naive recipients of splenocytes from UV irradiated mice treated with 0%, 0.1%, 0.5%, or 1.0% niacin supplementation, respectively, demonstrating niacin prevention of immunosuppression. Niacin supplementation elevated skin NAD content, which is known to modulate the function of DNA strand scission surveillance proteins p53 and poly(ADP-ribose) polymerase, two proteins critical in cellular responses to UV-induced DNA damage. These results clearly demonstrate a dose-dependent preventive effect of oral niacin on photocarcinogenesis and photoimmunosuppression and establish the capacity of oral niacin to elevate skin NAD levels. PMID- 10453440 TI - Low doses of diallyl disulfide, a compound derived from garlic, increase tissue activities of quinone reductase and glutathione transferase in the gastrointestinal tract of the rat. AB - Diallyl disulfide (DADS), a substance that is formed from the organosulfur compounds present in garlic, is known to increase tissue activities of the phase II detoxification enzymes quinone reductase (QR) and glutathione transferase (GT) in animals. In previous experiments, however, high doses of DADS were employed and only a limited range of tissues were examined. In the present studies, increased activities of QR and GT were recorded in the forestomach, glandular stomach, duodenum, jejunum, ileum, cecum, colon, liver, kidneys, spleen, heart, lungs, and urinary bladder of rats given DADS over a wide range of dose levels. Large variations in response were recorded among the different organs, with forestomach, duodenum, and jejunum being the most sensitive to enzyme induction by DADS. In these organs, significant increases in QR activity were observed at a dose of only 0.3 mg/kg/day. Such a dose level is close to that which may be achieved through human consumption of garlic, suggesting that induction of phase II enzymes may contribute to the protection that is afforded by this vegetable against cancer of the gastrointestinal tract in humans. PMID- 10453441 TI - Consumption of plant foods and stomach cancer mortality in the seven countries study. Is grain consumption a risk factor? Seven Countries Study Research Group. AB - Plant foods are generally considered to be beneficial for health. A higher consumption of fruits, and to a lesser extent vegetables, is consistently associated with a lower risk of stomach cancer. Results on the association between stomach cancer and grain consumption are less clear. We associated plant food consumption with 25-year stomach cancer mortality at population level in the Seven Countries Study. Around 1960, > 12,000 men aged 40-59 years from 7 countries and 16 cohorts were enrolled. In each cohort, dietary information was collected in small random samples. Crude and adjusted associations were calculated for a change of 10% of mean intake. Results differed for the plant foods studied: an inverse association was observed for fruits (adjusted rate ratio = 0.96, 95% confidence interval = 0.91-0.99), a positive relation for refined grains (adjusted rate ratio = 1.07, 95% confidence interval = 1.03-1.12), and no association for total plant foods, vegetables, whole grains, and potatoes. A high intake of refined grains was correlated with a low consumption of fruits. In conclusion, high intake of refined grains may increase stomach cancer risk. However, because adjustment could only be limited in this study, high intake of refined grains may just reflect the deleterious effect of a diet low in fruits or other characteristics associated with low fruit consumption. PMID- 10453442 TI - Plasma antioxidant vitamins and carotenoids in five Japanese populations with varied mortality from gastric cancer. AB - To examine the geographic associations between plasma antioxidant levels and gastric cancer risk, we conducted an ecological study in five regions of Japan representing the threefold variation in the disease mortality within the country. Subjects were 634 men aged 40-49 years sampled randomly from the five regions with 72% response rates. Plasma concentrations of five carotenoids (beta carotene, alpha-carotene, lycopene, lutein, and zeaxanthin), alpha-tocopherol, and ascorbic acid were measured, and the mean levels were correlated with age adjusted mortality rates from gastric cancer. beta-Carotene and alpha-tocopherol were inversely correlated with gastric cancer rates (r = -0.31 and -0.89, respectively). alpha-Carotene and lycopene showed stronger inverse correlation than did beta-carotene (r = -0.67 and -0.56, respectively), but these relations disappeared after the exclusion of one outlying region in Okinawa with the lowest mortality. In contrast, ascorbic acid revealed a negative correlation with the exclusion of this outlier (r = -0.61). Lutein and zeaxanthin were not inversely associated with risk. The results suggest that plasma levels of beta-carotene and alpha-tocopherol, and possibly alpha-carotene, lycopene, and ascorbic acid, may partly account for the regional difference in gastric cancer mortality in Japan. PMID- 10453443 TI - Phase I/II study of stage III and IV non-small cell lung cancer patients taking a specific dietary supplement. AB - This phase I/II study evaluates the influence of selected vegetables (SV) that contain known antitumor components on the survival of stage III-IV non-small cell lung cancer (NSCLC) patients. All patients were treated with conventional therapies. SV was added to the daily diet of 5 stage I patients in the toxicity study group (TG) and 6 stage III and IV patients in the treatment group (SVG), but not to the diet of 13 stage III and IV patients in the control group (CG). Age, Karnofsky performance status (KPS), and body mass index of SVG and CG patients were comparable at entry. KPS declined in the CG patients (79 +/- 8 to 55 +/- 11) but improved in the SVG patients (75 +/- 8 to 80 +/- 13) one to three months after entry. Weight change in the CG, SVG, and TG patients was -12 +/- 5%, -2 +/- 2%, and +4 +/- 4%, respectively. The median survival time and mean survival of the CG patients were 4 and 4.8 months, but in the SVG patients they were 15.5 and 15 months (p < 0.01). No clinical signs of toxicity were found in the TG patients in the 24-month study period. Adding SV to the daily diet of NSCLC patients was found to be nontoxic and associated with improved weight maintenance, KPS, and survival of stage III and IV NSCLC patients. PMID- 10453444 TI - Comparison of the 60- and 100-item NCI-block questionnaires with validation data. AB - Large epidemiological studies often require short food frequency questionnaires (FFQ) to minimize the respondent burden or to control for confounding from dietary factors. In this analysis, we compared the extensively used National Cancer Institute-Block 60- and 100-item FFQs with one another and with usual intake as estimated from 12 days of validation data. The analysis focused on nutrients from different aspects of the diet, including energy, fat, saturated fat, beta-carotene, dietary fiber, and vitamin C. By use of an errors-in variables analysis, the correlations of usual intake with the two types of FFQs for these nutrients were not different. Attenuation coefficients, a measure of misclassification error, for both FFQs were of similar magnitude and indicated that substantial attenuation of logistic regression coefficients would result from either FFQ. Our results confirm previous analyses describing the validity and utility of the 60-item FFQ (Epidemiology 1, 58-64, 1990) and indicate that it is essentially equivalent to the 100-item FFQ for epidemiological analyses of major nutrients. PMID- 10453445 TI - Subcellular accumulation of beta-carotene and retinoids in growth-inhibited NCI H69 small cell lung cancer cells. AB - Delivery of beta-carotene in tetrahydrofuran slowed the growth of NCI-H69 small cell lung cancer cells. Analysis of cells and cellular fractions revealed that beta-carotene-treated cells accumulated beta-carotene as well as some polar metabolites, primarily in the crude nuclei. Cells were grown at 1 x 10(5) cells/ml and treated with 20 microM beta-carotene. Growth monitoring up to 15 days indicated an inverse relationship between the duration of beta-carotene treatment and the rate of cell growth. Reverse-phase high-performance liquid chromatography analysis of treated cells showed the presence of beta-carotene, retinoic acid, retinol, and retinal, with beta-carotene accounting for the major material recovered. When cellular fractions were analyzed for beta-carotene, it was found to be located primarily in the crude nuclei. These results demonstrate that treatment of small cell lung cancer cells with beta-carotene results in a reduced growth of the cells. Further investigation is required to show a direct effect of beta-carotene or its intracellular polar metabolites on these cells. Accumulation of beta-carotene in the nucleus suggests a need for evaluating the nuclear role for beta-carotene. PMID- 10453446 TI - Consumption of green tea causes rapid increase in plasma antioxidant power in humans. AB - Green tea contains polyphenolic antioxidants that have shown anticarcinogenic properties in animal and in vitro experimental studies. Current data regarding absorption and bioavailability of tea antioxidants in humans, however, are conflicting. In this study, plasma and urine antioxidant power after ingestion of green tea was measured using the ferric reducing/antioxidant power (FRAP) assay (US patent pending) to assess absorption, systemic distribution, and renal excretion of green tea antioxidants in healthy adults. Results showed that absorption of green tea antioxidants was rapid, with peak increase in plasma FRAP of around 4% at 40 minutes after ingestion: mean increase was 44 +/- 9 (SE) mumol/l. Excretion of polyphenolic antioxidants was also fast, peaking at 60-90 minutes, with significant correlation between urinary FRAP values and urinary total phenolic concentrations (r = 0.845, p < 0.001). In control studies, no increase in plasma or urine FRAP values was seen after intake of water. Although the amount of antioxidants absorbed was relatively small and the increase in plasma antioxidant power was of short duration, results demonstrate that some potentially anticarcinogenic polyphenolic antioxidants in green tea enter the systemic circulation soon after ingestion and cause a significant increase in plasma antioxidant status. This increase may, in turn, lower oxidative damage to DNA and so decrease risk of cancer. PMID- 10453447 TI - Quercetin-induced apoptosis in colorectal tumor cells: possible role of EGF receptor signaling. AB - Flavonoids are among the best candidates for mediating the protective effect of diets rich in fruits and vegetables with respect to colorectal cancer. To gain additional information about their growth effects on colorectal tumors and their cellular mechanisms of action, a series of related flavonoids was added to cultures of colonic tumor cells. Most compounds induced growth inhibition and cell loss at concentrations of 1-100 microM, relative effectivity being quercetin > apigenin > fisetin > robinetin and kaempferol. Myricetin was only slightly effective. Quercetin was the strongest inducer of apoptosis in a process that was reversible until 10 hours by flavonoid removal and until 24 hours by fetal calf serum. Cells were preferentially retained in the S phase. On the cellular level, quercetin sensitivity was correlated with epidermal growth factor (EGF) receptor levels, rapid growth, and poor differentiation, indicating the possibility of targeting those cells most harmful for the organism. The flavonoid transiently inhibited EGF receptor phosphorylation but had only little effect on other signaling molecules. Even after recovery of receptor phosphorylation, cells remained resistant to EGF stimulation. In summary, the data indicate that inhibition of EGF receptor kinase is an integral part of quercetin-induced growth inhibition, but additional mechanisms also contribute to the overall effect. PMID- 10453448 TI - Dietary antioxidants and lung cancer risk: a case-control study in Uruguay. AB - To examine the protective role of dietary antioxidants (carotenoids, vitamin C, vitamin E, glutathione, and flavonoids) in lung cancer risk, a case-control study involving 541 cases of lung cancer and 540 hospitalized controls was carried out in Uruguay. The relevant variables were energy adjusted using the residuals method and then categorized in quartiles. Adjusted odds ratios (ORs) for antioxidants were calculated through unconditional logistic regression. With the exception of lycopene and vitamin C, the remaining antioxidants were associated with significant reductions in risk of lung cancer. Of particular interest was the inverse association between dietary glutathione and lung cancer [OR of quartile with highest intake compared with lowest quartile = 0.42, 95% confidence interval (CI) = 0.27-0.63]. Also, carotenoids and vitamin E were associated with significant reductions in risk of lung cancer (OR = 0.43, 95% CI = 0.29-0.64 for total carotenoids and OR = 0.50, 95% CI = 0.39-0.85 for vitamin E). A joint effect for high vs. low intakes of beta-carotene and glutathione was associated with a significant reduction in risk (OR = 0.32, 95% CI = 0.22-0.46). It could be concluded that dietary antioxidants are associated with a significant protective effect in lung carcinogenesis and that the inverse association for glutathione persisted after controlling for total vegetables and fruits. PMID- 10453449 TI - Childhood cancer in relation to cured meat intake: review of the epidemiological evidence. AB - Over the past two decades a series of epidemiological studies have examined the relationship between consumption of cured meats during pregnancy and the subsequent risk of brain tumors, as well as other cancers, in the offspring. The research was prompted in large part by experimental investigations showing that transplacental exposure to certain N-nitroso compounds, i.e., nitrosoureas, could produce brain tumors in laboratory animals. Fourteen such epidemiological studies, 13 of which used the case-control approach, are reviewed here. Most of the studies showed no significant association between total cured meat intake and childhood cancer risk but more found positive than negative relationships. Furthermore, several studies reported significant positive associations for maternal and sometimes childhood or paternal consumption of one or more cured meats, with odds ratios of twofold or greater reported among the highest consumers. On the other hand, a correlation analysis found no positive concordance between temporal trends from the 1970s to 1990s in childhood brain cancer rates and cured meat consumption, inasmuch as cancer rates rose over time while residual nitrite levels in cured meats fell sharply. Because of the potential for bias, especially recall bias, and/or confounding, the relatively weak magnitude of the associations reported, and the inconsistency between study findings, at this time it cannot be concluded that eating cured meat has increased the risk of childhood brain cancer or any other cancers. Moreover, although N-nitroso compounds are sometimes found in cured meats or may be formed endogenously, there is no empirical evidence that eating cured meats results in human neural nitrosourea exposure. Nevertheless, the hypothesis that eating nitrite-cured meats may influence childhood and perhaps adult brain cancer cannot be dismissed. Unbiased evaluation of the hypothesis may derive from the conduct of cohort studies, where the interview-derived information on cured meat intake precedes, or is not otherwise associated with, the diagnosis of cancer. PMID- 10453450 TI - Food coping strategies: a century on from Rowntree. AB - The objectives of this study were to update Rowntree's food estimates to bring them into line with contemporary food habits and changing lifestyles; to determine whether food budgeting/ coping strategies of low income households observed by Rowntree at the turn of the century were still prevalent in low income groups today: and to evaluate whether expenditure by low income households on food was sufficient to meet minimum dietary recommendations. Two hundred questionnaires were completed face to face with the interviewer within the Merseyside boundary and included only those households who were in the Post code area. Many low income households are unable to access a healthier diet due to income limitations. Many used a variety of coping strategies to stretch their incomes and were also found to rely on help from outside the immediate family such as extended family and friends and voluntary agencies. Households with children were found to be spending slightly more on food than the reconstructed Rowntree minimum dietary. There are thus two issues; firstly that there are households spending slightly more than the minimum yet that are unable to achieve a healthy diet. Secondly all the low income households are spending much less per person than the average person in the United Kingdom. It is concluded that whilst sterling work is being achieved within health promotion departments and within community health settings there is still a great deal to be undertaken. The value of Home Economics teaching in schools should be better recognised, and Home Economics should be reinstated in the National Curriculum. Secondly it is recommended that a standard approach be adopted to inform both welfare benefit payments and in-work top up payments, thus allowing households on low incomes to make informed choices within a budget that will accommodate healthier eating strategies. PMID- 10453451 TI - Dietary and lifestyle patterns in the aetiology of cataracts in Nigerian patients. AB - The objective of the study was to assess the dietary and lifestyle habits of patients with cataracts. A food frequency questionnaire was used to collect information on the consumption of a wide variety of food items especially fruits, vegetables and animal and dairy products. A structured questionnaire was used to collect information on smoking and alcohol consumption habits. The study was conducted on 62 subjects made up of 31 patients with cataracts attending the Ophthalmology clinic of Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Nigeria and 31 control subjects without cataracts. There were 20 males and 11 females in each group. The study showed that higher percentages of controls than patients had adequate intakes of fruits and vegetables. Vitamin supplement usage was also higher in controls than patients. There was a strong negative association between past history of smoking, alcohol consumption and cataract. We could not demonstrate a relationship between body mass index (BMI) and cataract in the present study. PMID- 10453452 TI - The relationship between habits of food consumption and reported reactions to food in people with inflammatory bowel disease--testing the limits. AB - Anecdotally, people with inflammatory bowel disease (IBD) report that some foods make them feel better and some foods make them feel worse. This paper reports about the relationship between food habits and reactions to foods in people with IBD. A database was designed to incorporate the date of data collection, the IBD disease, food habits, the quantity consumed and a response rating of the subjects' reactions to each food. For each of 122 foods, subjects were asked simultaneously about these five categories. Thirty three subjects with Crohn's disease and 27 with ulcerative colitis completed the questionnaire in two seasons. The focus of the examination was directed to the upper and lower limit of reactions to foods. Foods that affect people negatively and foods that affect people positively were identified in relation to habits of consumption. Foods that affect people negatively were among those never consumed while foods that affect people positively were among the regularly consumed foods. Findings included an increased number of foods that caused problems for people with Crohn's disease when compared with people with ulcerative colitis. Problems with chocolate, dairy products, fats and artificial sweeteners were documented in both groups of IBD. A standard food list generated without the input of individuals with a chronic illness may be inadequate to collect data about their consumption since many of the foods on the list were reportedly never consumed and there may be other foods not on the list that are consumed by this chronically ill population. PMID- 10453453 TI - What should be done to compensate for the shortcomings of a vegetarian diet? PMID- 10453454 TI - Why is so much good dietary advice being ignored? PMID- 10453455 TI - Encouraging healthy diets in adolescent girls. PMID- 10453456 TI - Is organic food production feasible? AB - The problems of modern agriculture and whether organic methods of food production offer a practical solution in modern society are reviewed and discussed, with frank admission of drawbacks in organic methods. PMID- 10453457 TI - The 'chicken and egg' problem of co-evolution of peptides and their cognate receptors: which came first? PMID- 10453458 TI - Thyrotropin releasing hormone (TRH), the TRH-receptor and the TRH-degrading ectoenzyme; three elements of a peptidergic signalling system. PMID- 10453459 TI - Corticotropin-releasing factor (CRF) and its role in the central nervous system. PMID- 10453460 TI - CRF and CRF receptors. PMID- 10453461 TI - Neural oxytocinergic systems as genomic targets for hormones and as modulators of hormone-dependent behaviors. AB - At the molecular level, estradiol turns on the gene for oxytocin in a subset of paraventricular hypothalamic neurons and turns on the gene for the oxytocin receptor in other limbic and hypothalamic cell groups. As a result, oxytocin deposition, whose signal is transduced both through G alpha (q/11) and Gi to stimulate phosphatidylinositol turnover, facilitates electrical activity in certain hypothalamic neurons. Consequently, affiliative behaviors including those closely associated with reproduction--mating behaviors and parental behaviors- are promoted. One important aspect of this effect is the preservation of instinctive behaviors associated with reproduction, in the face of disturbances due to mild stress. PMID- 10453462 TI - Vasopressin receptors: structural functional relationships and role in neural and endocrine regulation. PMID- 10453463 TI - The oxytocin receptor. PMID- 10453464 TI - Targeted mutagenesis of the murine opioid system. PMID- 10453465 TI - Orphan receptors and the concept of reverse physiology: discovery of the novel neuropeptide orphanin FQ/nociceptin. AB - The cloning of numerous orphan members from the supergene family of G protein coupled receptors implies the existence of many as yet undiscovered neurotransmitters and neuropeptides. Recently, new technologies were developed to isolate natural ligands for orphan receptors, using the receptor as a biological sensor during the purification process. This manuscript will present the concept and technology of an approach which starts from a cloned receptor to ultimately describe the physiological functions of the transmitter system. This strategy inverts the classical order of biomedical research and was thus termed "reverse physiology". The first natural ligand isolated by this strategy is a peptide with significant similarity to the opioid peptides and has been named orphanin FQ or nociceptin (OFQ/NOC). Evidence for characterizing OFQ/NOC as a genuine neuropeptide will be reviewed. OFQ/NOC is biosynthetically derived from a larger precursor protein which may encode additional bioactive peptides. Since its discovery, a large number of studies have described numerous physiological functions of OFQ/NOC. Because of its relation to the opioid system, much attention has been focused on the involvement of OFQ/NOC in nociception, sometimes with controversial results. However, the pharmacological profile of the OFQ/NOC system suggests a clear separation from the opioids. The discovery of OFQ/NOC and the subsequent analyses of its physiological functions is an example which has already been followed by the identification of two other novel neuropeptides. The orphan receptor strategy holds a lot of promises for the postgenomic era, helping to fill the vast amount of sequence data with life. PMID- 10453466 TI - Molecular biology of the receptors for somatostatin and cortistatin. PMID- 10453467 TI - Novel neurotransmitters for sleep and energy homeostasis. AB - We have developed methodologies for identifying mRNAs with highly restricted expression within the brain. One postnatal-onset mRNA, restricted to sparse GABAergic interneurons of the cerebral cortex and hippocampus, encodes preprocortistatin, the precursor of a 14-residue peptide that shares 11 amino acids with somatostatin. Cortistatin binds to all five cloned somatostatin receptors when they are expressed in transfected cells and depresses neuronal activity, but, unlike somatostatin, it reduces locomotor activity and induces slow-wave sleep. Cortistatin, whose mRNA accumulates during sleep deprivation, apparently acts by antagonizing the effects of acetylcholine on cortical excitability, thereby causing synchronization brain slow waves. A single amino acid difference with somatostatin accounts for the dramatic differences in the effects of the two peptides on physiology and behavior. A second postnatal-onset mRNA, restricted to 1100 large neuronal cell bodies of the dorsal-lateral hypothalamus, encodes preprohypocretin, the precursor of two peptides that share homology with each other and with members of the secretin peptide family. The peptides are detected immunohistochemically in secretory vesicles at synapses of fibers that project to posterior hypothalamus and diverse targets in other brain regions. The peptides are excitatory when applied to cultured hypothalamic neurons. Recent studies by Sakurai and colleagues (1998) have identified the hypocretin peptides (called the orexins by those workers) as ligands for two orphan receptors at which they stimulate food-intake behavior. Sakurai and collaborators showed that the mRNA for these peptides accumulates during food deprivation. The hypocretin projections suggest additional homeostatic roles for the peptides. These studies suggest the common mechanism of regulation for necessary, but voluntary, behaviors (sleep and feeding) by transcription-based accumulation of peptide transmitters that create a pressure for the voluntary activities. PMID- 10453468 TI - Galanin and galanin receptors. AB - The development of a strain of galanin knockout mice has provided confirmation of a neuroendocrine role for galanin, as well as supporting results of previous physiological investigations indicating a role for galanin in analgesia and neuropathic pain, and potentially in neuronal growth and regeneration processes. Whether elevation of galanin expression in neurodegenerative disorders such as Alzheimer's disease represents a survival response or exacerbates functional deficit in afflicted individuals remains to be determined. More detailed analysis of the phenotype of the galanin knockout mouse should provide insights into the physiological role of galanin in memory and learning processes, as well as in hypothalamic function and other aspects of neuroendocrine regulation. Biochemical and molecular cloning efforts have demonstrated that the multiplicity of actions of galanin is matched by complexity in the distribution and regulation of galanin and its receptors. A focus on characterisation of galanin receptors has resulted in the molecular cloning of three receptor subtypes to date. The distribution and functional properties of these receptors have not yet been fully elucidated, currently precluding assignment of discrete functions of galanin to any one receptor subtype. It is not currently possible to reconcile available pharmacological data using analogs of galanin and chimeric peptides in functional assay systems with the pharmacological properties of cloned receptor subtypes. This highlights the value of further knockout approaches targeting galanin receptor subtypes, but also raises the possibility of the existence of additional receptor subtypes that have yet to be cloned, or that receptor activity may be modulated by regulatory molecules that remain to be identified. The development of receptor subtype-specific compounds remains a high priority to advance work in this area. The ability to selectively modulate the many different actions of galanin, through a clearer understanding of receptor structure-function relationships and neuronal distribution, promises to provide important insights into the molecular and cellular basis of galanin action in normal physiology, and may provide lead compounds with therapeutic application in the prevention and treatment of a range of disorders. PMID- 10453469 TI - The cholecystokinin--gastrin family of peptides and their receptors. PMID- 10453470 TI - Function of the neuropeptide head activator for early neural and neuroendocrine development. PMID- 10453471 TI - Invertebrate neurohormones and their receptors. PMID- 10453473 TI - Purification and characterization of the heat-labile alpha-amylase secreted by the psychrophilic bacterium TAC 240B. AB - A total of 59 bacteria samples from Antarctic sea water were collected and screened for their ability to produce alpha-amylase. The highest activity was recorded from an isolate identified as an Alteromonas species. The purified alpha amylase shows a molecular mass of about 50,000 Da and a pI of 5.2. The enzyme is stable from pH 7.5 to 9 and has a maximal activity at pH 7.5. Compared with other alpha-amylases from mesophiles and thermophiles, the "cold enzyme" displays a higher activity at low temperature and a lower stability at high temperature. The psychrophilic alpha-amylase requires both Cl- and Ca2+ for its amylolytic activity. Br- is also quite efficient as an allosteric effector. The comparison of the amino acid composition with those of other alpha-amylases from various organisms shows that the cold alpha-amylase has the lowest content in Arg and Pro residues. This could be involved in the principle used by the psychrophilic enzyme to adapt its molecular structure to the low temperature of the environment. PMID- 10453474 TI - Cresol metabolism by the sulfate-reducing bacterium Desulfotomaculum sp. strain Groll. AB - The metabolism of cresols under sulfate-reducing conditions was investigated in Desulfotomaculum sp. strain Groll. This strain grows on a variety of aromatic compounds, including para- and meta- but not ortho-cresol. Degradation of p cresol proceeded by oxidation reactions of the methyl group to yield p hydroxybenzoate, which was then dehydroxylated to benzoate. The aromatic intermediates expected for this pathway, p-hydroxybenzyl alcohol, p hydroxybenzaldehyde, p-hydroxybenzoate, and benzoate, were readily metabolized by strain Groll. Utilization of these intermediates generally preceded and inhibited the degradation of p-cresol. p-Hydroxybenzoate and benzoate were detected in culture fluid as metabolites of p-cresol. p-Hydroxybenzaldehyde and p hydroxybenzoate were detected in cultures degrading p-hydroxybenzyl alcohol. Enzyme activities responsible for utilization of p- and m-cresol, induced by growth on the respective cresol, were detected in cell-free extracts of strain Groll. The compounds detected in culture fluids and the enzyme activities detected in cell-free extracts indicate that the pathways for the degradation of p- and m-cresol converge on benzoate, followed by metabolism to benzoyl-coenzyme A (CoA). Strain Groll can utilize both cresol isomers under sulfate-reducing conditions by similar reactions, but the enzyme activities catalyzing these transformations of the two isomers appear distinct. PMID- 10453475 TI - Stress tolerance in a yeast lipid mutant: membrane lipids influence tolerance to heat and ethanol independently of heat shock proteins and trehalose. AB - The response of a yeast unsaturated fatty acid auxotroph, defective in delta 9 desaturase activity, to heat and ethanol stresses was examined. The most heat- and ethanol-tolerant cells had membranes enriched with oleic acid (C18:1), followed in order by cells enriched with linoleic (C18:2) and linolenic (C18:3) acids. Cells subjected to a heat shock (25-37 degrees C for 30 min) accumulated trehalose and synthesized typical heat shock proteins. Although there were no obvious differences in protein profiles attributable to lipid supplementation of the mutant, relative protein synthesis as determined by densitometric analysis of autoradiograms suggested that hsp expression was different. However, there was no consistent relationship between the synthesis of heat shock proteins and the acquisition of thermotolerance in the lipid supplemented auxotroph or related wild type. Furthermore, trehalose accumulation was also not closely related to stress tolerance. On the other hand, the data presented indicated a more consistent role for membrane lipid composition in stress tolerance than trehalose, heat shock proteins, or ergosterol. We suggest that the sensitivity of C18:3-enriched cells to heat and ethanol may be attributable to membrane damage associated with increases in membrane fluidity and oxygen-derived free radical attack of membrane lipids. PMID- 10453476 TI - Regulated expression of HIV-1 Rev function in mammalian cell lines. AB - In order to facilitate further investigation of Rev function, we have generated two systems for the inducible expression of Rev in mammalian cell lines (HeLa and U937) using either a tetracycline-regulated promoter or fusion of Rev to a modified form of the hormone binding domain of the estrogen receptor. In the case of the fusion of Rev to the modified hormone binding domain of the estrogen receptor, we demonstrated induction of Rev function in response to tamoxifen administration to levels comparable to that of the unmodified Rev protein. Subsequently, U937 lines were generated that retained the observed pattern of hormone-dependent function of the Rev fusion protein. In the case of the tetracycline-regulated system, cell lines (both HeLa and U937) were generated that displayed tight regulation of Rev. In the case of the HeLa cell lines, they were used for the subsequent generation of stable cell lines expressing either HIV-1 env or chloramphenicol acetyl transferase (CAT) in a Rev-dependent fashion. Using the latter cell lines, we demonstrate the ability to control Rev expression over a broad concentration range and find that, as soon as Rev expression is detectable, induction of Rev-dependent gene expression is also observed. PMID- 10453477 TI - Utilization of low molecular weight aromatic compounds by heterobasidiomycetous yeasts: taxonomic implications. AB - The utilization of low molecular weight aromatic compounds implies the operation of complex metabolic pathways. In order to investigate the taxonomic relevance of this property among heterobasidiomycetous yeasts, both at the species level and at higher taxonomic ranks, the capacity to assimilate twenty such compounds was tested in a total of 332 strains representing approximately 200 species. The substrates most frequently utilized were protocatechuic, caffeic, and p hydroxybenzoic acids, whereas cinnamic, sinapic, and syringic acids and guaiacol were never assimilated. The assimilation of the majority of the aromatic compounds investigated correlated with the utilization of protocatechuic acid. Among the Urediniomycetes, the members of the Sporidiales and those of the Naohidea-Rhodotorula minuta clade showed a good ability to utilize aromatic compounds, whereas the members of the Agaricostilbum-Kondoa group were more heterogeneous, in agreement with the four subclades known. Among the Tremellomycetidae, the members of the Cystofilobasidium and Tremella clades showed a reduced or null ability to utilize aromatic compounds. In contrast, the members of the Trichosporon clade were able to utilize phenol and similar substrates, and the representatives of the Filobasidium clade assimilated various aromatic compounds, including those requiring more complex catabolic routes. Assimilation tests using, as sole carbon and energy sources, low molecular weight aromatic compounds appear to be potentially useful in taxonomic studies of basidiomycetous yeasts. In those species in which a considerable number of strains was investigated, variable assimilation patterns were frequently observed. The possibility that such discrepant results indicate an incorrect species delimitation is discussed. PMID- 10453478 TI - Isolation and characterization of thermophilic bacteria capable of degrading dehydroabietic acid. AB - Using a semi-continuous enrichment method, we isolated two thermophilic bacterial strains, which could completely degrade abietane resin acids, including dehydroabietic acid (DhA). Strain DhA-73, isolated from a laboratory-scale bioreactor treating bleached kraft mill effluent at 55 degrees C, grew on DhA as sole carbon source; while DhA-71, isolated from municipal compost, required dilute tryptic soy broth for growth on DhA. DhA-71 grew on DhA from 30 degrees C to 60 degrees C with maximum growth at 50 degrees C; while, DhA-73 grew on DhA from 37 degrees C to 60 degrees C with maximum growth at 55 degrees C. At 55 degrees C, the doubling times for DhA-71 and DhA-73 were 3.3 and 3.7 h, respectively. DhA-71 and DhA-73 had growth yields of 0.26 and 0.19 g of protein per g of DhA, respectively. During growth on DhA, both strains converted DhA to CO2, biomass, and dissolved organic carbon. Analyses of the 16S-rDNA sequences of these two strains suggest that they belong to two new genera in the Rubrivivax subgroup of the beta subclass of the Proteobacteria. Strains DhA-71 and DhA-73 are the first two bacteria isolated and characterized that are capable of biodegradation of resin acids at high temperatures. This study provided direct evidence for biodegradation of resin acids and feasibility for biotreatment of pulp mill effluent at elevated temperatures. PMID- 10453479 TI - Cross-induction of pyrene and phenanthrene in a Mycobacterium sp. isolated from polycyclic aromatic hydrocarbon contaminated river sediments. AB - A polycyclic aromatic hydrocarbon (PAH)-degrading culture enriched from contaminated river sediments and a Mycobacterium sp. isolated from the enrichment were tested to investigate the possible synergistic and antagonistic interactions affecting the degradation of pyrene in the presence of low molecular weight PAHs. The Mycobacterium sp. was able to mineralize 63% of the added pyrene when it was present as a sole source of carbon and energy. When the enrichment culture and the isolated bacterium were exposed to phenanthrene, de novo protein synthesis was not required for the rapid mineralization of pyrene, which reached 52% in chloramphenicol-treated cultures and 44% in the absence of the protein inhibitor. In the presence of chloramphenicol, < 1% of the added pyrene was mineralized by the mixed culture after exposure to anthracene and naphthalene. These compounds did not inhibit pyrene utilization when present at the same time as pyrene. Concurrent mineralization of pyrene and phenanthrene after exposure to either compound was observed. Cross-acclimation between ring classes of PAHs may be a potentially important interaction influencing the biodegradation of aromatic compounds in contaminated environments. PMID- 10453480 TI - Subcellular localization of the Streptococcus mutans P1 protein C terminus. AB - To determine the subcellular location of the Streptococcus mutans P1 protein C terminal anchor, cell envelope fractionation experiments were conducted in combination with Western immunoblotting, using monoclonal antibody MAb 6-8C specific for an epitope that maps near the C terminus of P1 protein and also a polyclonal antibody preparation directed against the P1 C-terminal 144 amino acids (P1COOH). P1 protein was detected in cell walls but not the membrane purified from S. mutans cells by the monoclonal antibody. In contrast, P1 protein was not detected in the same cell wall preparation using the anti-P1COOH polyclonal antibody. However, proteins released from the cell walls by treatment with mutanolysin contained antigen that was recognized by the anti-P1COOH antibody, suggesting that the epitopes recognized by the antibody were masked by peptidoglycan in the cell wall preparations. When cell walls were treated with boiling trichloroacetic acid to solubilize cell-wall-associated carbohydrate, P1 antigen could not be detected in either the solubilized carbohydrate, or in the remaining peptidoglycan, regardless of whether polyclonal or monoclonal antibody was used. However, when the peptidoglycan was treated with mutanolysin, P1 antigen could be detected in the mutanolysin solubilized fraction by MAb 6-8C. Collectively, these data suggest that the C-terminal 144 amino acids of the P1 protein are embedded within the cell wall, and associated exclusively with the peptidoglycan. Furthermore, the ability of the anti-P1COOH antibody to recognize P1 antigen only after mutanolysin treatment of cell walls suggests these C terminal 144 amino acids are tightly intercalated within the peptidoglycan strands. PMID- 10453481 TI - Effects of grammatical gender on picture and word naming: evidence from German. AB - The present study investigated the effects of prior grammatical gender information (masculine, feminine, and neuter) provided by a minimal sentence context on both picture- and word-naming latencies. Named targets were nouns or pictures of concepts featuring unambiguous grammatical gender. Simple sentence fragments were presented auditorily prior to each picture or word target; the relation between these sentence primes and the word or picture target was either gender-congruent, gender-incongruent, or gender-neutral. Relative to the gender neutral baseline, reliable facilitation and inhibition effects were both observed in the picture-naming task. By contrast, only inhibition effects were observed in the word-naming task. The results suggest that the processes of picture and word naming differ qualitatively when gender information is precued. The findings are discussed with respect to the distinction between postlexical and intralexical loci of the effects of gender context on word recognition and production. PMID- 10453482 TI - Semantic influence on processing gender agreement: evidence from Hebrew. AB - In the present study we examined the influence of manipulating the animacy of the sentential subject on the size of the syntactic priming effect induced by violation of subject-predicate gender agreement in Hebrew. The agreement violation delayed naming incongruent compared with congruent predicates. This priming effect was stronger when the sentential subject was an animate than an inanimate noun. Additional experiments revealed that: (1) the interaction between the priming effect and animacy of the subject could not be explained on the basis of differences in the phonological transparency of the gender inflection in the two groups of nouns, and (2) it was sensitive to the ratio of animate/inanimate conditions in a block. We suggest that the interaction between the processing of agreement and the effect of animacy is influenced by a controlled process of verifying the coherence of a currently identified word within a built-up context. PMID- 10453483 TI - Changes in the phage typing patterns of Staphylococcus aureus strains at Concepcion, Chile, in the last 30 years. AB - Staphylococcus aureus is a ubiquitous pathogen still implicated as a common cause of community- and hospital-acquired infections. This micro-organism has demonstrated an immense adapting capacity to rapid environmental changes. In recent years, multiresistant strains have caused increasing nosocomial infections in several parts of the world. In the period 1993-94, 455 clinical isolates were typed on the basis of traditional phage typing procedure and these data were compared with others from similar studies carried out at the Department of Microbiology, University of Concepcion in 1960, 1972, and 1982. Throughout the years, phage groups have been shifting from group I to group III and examination of phage types show that types 80 and 80/81 which were the most virulent and resistant by the 1960s, had disappeared. Nowadays, types 75 and 54/75 are most frequently found, and these have been associated with methicillin-resistant S. aureus. PMID- 10453485 TI - High temperature induced antibiotic sensitivity changes in Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa, which was resistant to a wide variety of antibiotics, became sensitive to several of these antibiotics when grown and tested at 46 degrees C. Cell wall antibiotics such as penicillin G and ampicillin were only effective when added to cells growing at 46 degrees C prior to a temperature shift to 37 degrees C. Antibiotics which penetrate the cytoplasmic membrane to express their inhibiting action present a pattern different from those which are active against the outer cell wall. In order that these compounds be effective, the permeability of the cytoplasmic membrane must be further altered with agents such as EDTA which allow the penetration of actinomycin D. Inhibitors of protein synthesis, such as streptomycin and chloramphenicol, have increased access to their sites of action in cells grown at 46 degrees C. Cells grown at 46 degrees C have 40% less lipopolysaccharide (LPS) than cells grown at 37 degrees C and the LPS aggregates were of large molecular size in cells grown at 46 degrees C. Growth at 46 degrees C affects the permeability properties of the outer cell wall more than the permeability properties of the cytoplasmic membrane and this was due, in part, to the selective release of LPS of LPS-protein complexes at elevated growth temperatures. PMID- 10453484 TI - Vancomycin-resistant enterococci isolates from the rumen content of deer. AB - The total enumeration of enterococci from the rumen content of deer, their species determination, resistance to antibiotics with the predominant importance of vancomycin resistance, and their metabolic properties, were examined. Enterococci (2.0 x 10(2) CFU per ml) encountered from the rumen of deer were phenotypically allotted to the species Enterococcus casseliflavus (26.6%), E. gallinarum (26.6%) and E. faecium (6.6%). The other isolates were not phenotypically specified. Although the resistance to the other antibiotics was recorded, the emergence of vancomycin-resistant enterococci in a high percentage (60%) was revealed. The majority of strains (80%) were multiresistant with the predominant resistance shown by vancomycin. These strains produced lactic acid in the range from 0.258 mol l-1 up to 0.489 mol l-1, and most of the isolates (53.3%) manifested medium urease activity. Vancomycin-resistant enterococci were selected for other studies such as plasmid determination and bacteriocin production, and to assess their value as indicators of faecal contamination. PMID- 10453486 TI - [Construction of human genomic YAC clones with "terminal modification"]. AB - A human YAC genomic pool with more than 4,000 clones was constructed by means of "terminal modification" of YAC vector and genomic inserts, as well as optimization of relevant experimental parameters. The transformation rate is approximately 400-500 transformants per microgram DNA. Preliminary characterization of randomly sampled clones demonstrates that YAC-specific inserts, in the average size of 450 kb and ranging 150-750 kb, were detected from 80% of the clones. The vector ligation experiment indicates the efficient reduction of "coligation" by terminal modification. PMID- 10453487 TI - [The distribution of motoneuronotrophic factor 1 (MNTF1) and its receptor-like substance in the spinal cord and limb muscles of mice with motoneuron disease]. AB - Using a monoclonal antibody for the motoneuronotrophic factor 1 (MNTF1) and its idiotypic antibody we have detected immunohistochemically the changes in the expression of the positive immunoreactivities for the MNTF1 and its receptor-like substance in the cervical and lumbar spinal cord regions as well as in the limb muscles of the wobbler mice, as the disease progressed from the first to the fourth stage. The results indicated that the positive immunoreactivities for the MNTF1 and its receptor-like substance were mainly located in the soma and neurites of the anterior born motoneurons at spinal layers VIII and IX and in the sarcoplasm of the skeletal muscles of the fore- and hind-limbs, and whereas their nuclei were immunoreactively negative. In the first stage (3 weeks) of the disease there were fewer in numbers of cells containing positive immunoreactivities (P < 0.01) and lesser in their relative amount of immunoreactivities per cell (P < 0.01) in the cervical and lumbar spinal cord as well as in the fore- and hind-limbs muscles of the wobbler mice as compared to those of the normal littermates. In the fourth stage (3 months) of the disease these differences between the wobbler mice and the normal littermates were even more distinct (P < 0.01 and P < 0.01). These results suggested that as syntheses of the skeletal muscle target cell-derived MNTF1 and its receptor-like substance were reduced to levels which were not enough to ensure the survival of motoneurons and consequently leading to muscular atrophy, as the motoneuron disease in wobbler mice deteriated. PMID- 10453488 TI - [Effect of L-arg and SNP on pulmonary arterial pressure and vascular structural changes of chronically hypoxic rats]. AB - The effects of L-arginine (L-arg) and sodium nitroprusside (SNP) on pulmonary arterial pressure, the percentage of muscularization of intra-acinar vessels and ultrastructural changes of extra pulmonary artery and pulmonary arteriole of chronically hypoxic rats were studied. The results showed that: (1) Both L-arg and SNP decreased mean pulmonary arterial pressure of chronically hypoxic rats significantly. (2) Both L-arg and SNP reduced the percentage of muscularization of intra-acinar vessels of chronically hypoxic rats significantly. (3) Both L-arg and SNP protected pulmonary artery from the damages of endothelium and the changes in smooth muscle cell phenotype by hypoxia. These results suggested that exogenous nitric oxide might play a role in the protection of pulmonary arterial function and structure which alleviate the development of pulmonary hypertension induced by chronic hypoxia. PMID- 10453489 TI - [Comparison of radiographs and magnetic resonance imaging in the detection of sacroiliitis in patients with ankylosing spondylitis]. AB - The role of radiography and magnetic resonance (MR) imaging in the detection of sacroiliitis in patients with ankylosing spondylitis (AS) was compared in this study. Thirty-six sacroiliac joints in 18 patients with AS were examined with radiography and MR scan. MR images were performed with the sequence of the coronal T1 weighted image, T2 weighted image, and T2* weighted image. Images of all patients were graded according to the modified New York criteria. Statistical results showed that significant differences existed between MR imaging and radiography in the detection of sacroiliitis (P < 0.01). MR imaging was superior to plain film radiographs in visualising erosions (P < 0.01). Radiography cannot reveal the cartilage changes and bone marrow oedema, which can only be seen in MR images. In the absence of radiographic changes, MR imaging can provide objective and complementary findings of sacroiliitis in patients with AS. Due to the ability to image cartilage changes and bone marrow oedema directly, MR imaging may be particularly useful in early diagnosis of sacroiliitis. For patients without any changes on sacroiliac joint radiograph, further examination of MR imaging may be advisable. PMID- 10453491 TI - [Cloning of genes associated with taxol-induced apoptosis of human breast cancer cells]. AB - The method of mRNA differential display was employed to isolate expressing genes involved in taxol-induced apoptosis of human breast cancer cells (BCap 37). Results demonstrated that the expressive alteration of twelve clones isolated was associated with taxol-induced apoptosis identified by Northern blot. cDNA sequence of clone C3P3 was highly homologous (99%) to human S-adenosylmethionine (S-AdoMet) synthetase. Increased transcriptional level of clone C3P3 induced by taxol was parallel to enhanced enzyme activity of S-AdoMet synthetase. These results suggested that the expression of these clones isolated by the mRNA differential display be possibly involved in the regulation of taxol-induced apoptosis. Their function in details need further study. PMID- 10453490 TI - [Effect of recombinant growth hormone on amino acids metabolism in blood and urine after major operation]. AB - To determine the effects of recombinant human growth hormone (GH) on postoperative amino acids metabolism, we performed a placebo-controlled randomized double-blind trial in 18 patients after elective gastrectomy or colectomy. The same amount of calories and amino acids were administered. Research group was daily injected growth hormone 0.15 IU/(kg.day) and control group was given placebo during the first postoperative week. All urine were analyzed for nitrogen balance and excretion of 3-methylhistidine. On the seventh postoperative day while the parenteral nutrients were being infused, amino acid exchange across the forearm was measured. On the seventh postoperative day all patients' amino acids in plasma were slightly increased by comparing with preoperative day but was not statistical significance. Forearm arterial-venous difference (A-V) measured on POD + 7 was markedly different between groups. There was continued release of amino acids from the forearm in the control subjects, while there was marked amino acid uptake in the GH patients (total amino acid nitrogen exchange was -398 +/- 158 mumol/L vs +165 +/- 61 mumol/L, P < 0.01). Excretion of urinary 3-methylhistidine in control group loss more than in GH group (7.3 +/- 1.0 mumol/(kg.24 h) vs 4.4 +/- 0.3 mumol/(kg.24 h), P < 0.05). The results demonstrated that GH can increase the uptake of amino acid from forearm and decrease the excreation of 3-methylhistidine in urine after operation, but did not disturb the balance of amino acids in plasma. PMID- 10453492 TI - [Limitations of the common pre-processing method in the signal-averaged electrocardiogram]. AB - The isolation of the micro potentials from electrocardiogram using coherence averaging technique requires perfect signal alignment. The purpose of this paper was to assess the reliability of the maximum correlation coefficient (MCC) method by computer simulation. The ability of signal recognition, the impacts of correlation-window-width and threshold on accuracy of temporal alignment and on averaged results were analyzed. The results showed that the MCC method was totally insensitive to amplitude variation and relatively insensitive to the duration variation. In the process of signal alignment, the narrower correlation window-width, the less sensitivity of the maximum correlation coefficient to the misalignment would be. With the threshold of 0.99, the window-width of both 40 and 100 ms induced the deformation on averaged result in time and frequency domain. The simulation results suggested that the deficiencies of MCC method could impose serious limitations on reliability of isolating the atrial and ventricular late potentials. PMID- 10453493 TI - [Effects of captopril on the myocardial performances and hemodynamics after cardiac arrest]. AB - The effects of cardioplegia and reperfusional blood containing 4.6 mumol/L captopril (CPL) on the myocardial performances and hemodynamics of canine heart under hypothermic extracorporeal circulation were studied. Twenty four mongrel dogs were divided randomly into 3 groups. CONTROL GROUP: Only modified St. Thomas cardioplegia was used. CPL group: St. Thomas cardioplegia containing CPL 4.6 mumol/L was used and the same concentration of captopril in the circulatory blood was established before releasing the cross clamp. CPL + phenylephrine group: The concentration was the same as the CPL group, with an additional dose of phenylephrine (2 micrograms/kg) into the oxygenator and abdominal coarctation. The results revealed that the coronary blood flow and left ventricular contractility and relaxation indices of CPL group were significantly improved than those of the control group, and increasing the systemic and pulmonary vascular resistence had no effect on the cardiac function protected by captopril. It is suggested that captopril could ameliorate the recovery of myocardial performences by inhibitting myocardial and systemic renin-angiotensin system and scavenging free radicals. PMID- 10453494 TI - Effects of superoxide anion, B(alpha)P and TPA on the membrane fluidity of NIH3T3 cells. AB - The membrane fluidity of NIH3T3 cells treated with low and high concentration of cell stimulatives (extracellular generated superoxide anion(O2-.),12-O-tetra decanoyl-phorbol-13-acetate(TPA), and benzo(alpha)-pyrene[B(alpha)P]) was investigated by means of fluorescence labels 1,6-diphenyl-1,3,5 hexatriene (DPH) and N-(3-pyrene) maleimide (N(3p)M). The high concentration of O2-., TPA, B(alpha)P greatly increased the fluidity of cell membrane lipid domain. No changes of the florescence polarization of DPH was found in membrane lipid domain treated with low concentration of O2-., TPA, and B(alpha)P. However, decrease in the fluoresence polarization of N(3p)M on the cell membrane protein domain damaged by low concentration of cell stimulatives was observed, showing that these treatment could influence the conformation of membrane protein. The possible relationship between the changes of the conformation of membrane protein and the cell transformation and its carcinogenic machenisms were discussed. PMID- 10453495 TI - [Effects of the glucoprotein component of musk on the functions of rat polymorphonuclear leukocytes activated by fMLP in vitro]. AB - To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on the release of superoxide anion as well as beta glucuronidase and lysozyme of rat polymorphonuclear leukocytes activated by fMLP. An in vitro incubation system with rat polymorphonuclear leukocytes was used. Superoxide anion production was determined by cytochrome C reduction. beta glucuronidase and lysozyme release was quantitated by enzyme reactions in which phenolphthaleinglucuronic acid and micrococcus lysodeikticus were as the substrates, respectively. In comparison with control, musk-1 at final concentrations of 1-100 micrograms/ml can increase superoxide anion production by 23.0%-83.6% and decrease beta-glucuronidase and lysozyme release by 7%-47% and 9% 22%, respectively, in rat polymorphonuclear leukocytes. It is concluded that Musk 1 can significantly affect the functions of rat polymorphonuclear leukocytes. Therefore, inhibition of lysosomal enzyme release might be considered as one of the mechanisms of anti-inflammatory role of musk. PMID- 10453497 TI - [Study on the effect of ofloxacin and tanshinone II A on human leukocyte chemotactic migration in vitro]. AB - In order to verify the possible anti-inflammatory activity and their potency of ofloxacin and Tanshinone II a, the effects of different concentrations of ofloxacin, Tanshinone II a and hydrocortisone on human leukocyte migration in vitro were investigated employing agarose migration technique in which f-Met-Lea Phe was served as chemotactic factor. The results showed that all the three agents could inhibit migration of leukocytes remarkably. The effects were concentration-dependent with excellent linear relationship. The IC50 were: Tanshinone II a 0.46 microgram/ml, ofloxacin 0.63 microgram/ml, and hydrocortisone 46.52 micrograms/ml respectively. The results indicated that Tanshinone II a was the most potent agent for inhibiting migration of human leukocytes in vitro and the hydrocortisone was relatively less potent. PMID- 10453496 TI - [Expression of IL-4 and IL-10 by PBMC and tumor tissues in cancer patients]. AB - Using biological activity assay, the production of IL-4 and IL-10 by peripheral blood mononuclear cells (PBMC) was studied in patients with nasopharyngeal carcinoma and lymphoma. The IL-2, IL-4 and IFN gamma levels in both cancer patients were lower than that in normal subjects. In contrast, the IL-10 level was higher in both cancer patients. When rIL-2 was added into the cytokine production system, IL-2, IL-4, and IFN gamma level were upregulated in lymphoma patients. However, in the patients of nasopharyneal carcinoma, only IL-2 level was upregulated. IL-10 activity could not be affected by exogenous IL-2 in both cancer patients. The results of negative IL-4 activity and higher positive percentage (84.6%) of IL-10 activity in ascites from patients with ovarian carcinoma were corresponded to the patients with nasopharyngeal carcinoma and lymphoma. By using in situ hybridization technique, the expression of IL-4 and IL 10 was studied in tumor tissues from patients with gastric, esophageal and breast carcinomas. There was no differences of the percentage of IL-4 and IL-10 expression among three kinds of tumor tissues, the range of positive percentage was 40%-70%. These results indicate that the correction of abnormal upregulation of IL-10 activity should be considered for cancer immunotherapy. PMID- 10453498 TI - [The setting up of human serum calcitonin radioimmunoassay and its clinical application]. AB - This study deals with the setting up of human serum calcitonin radioimmunoassay (hCT-RIA) using the high titer and specific antibody which was prepared by our own laboratory. The final dilution of the antibody titer was 1:1,200,000 when the binding rate of Bo/T was 30%. The antibody's affinity constant (Kd) was 2.25 x 10(11) L/M. No cross reaction was found between the antibody and the following eight different peptide hormones and proteins, viz.: TSH, T3, T4, ACTH, PTH, BGP, Insulin and Gastrin. The coefficients variation of intra and inter assay were 3.2% and 9.0%, respectively. The mean recovery rate of CT was 99.8%. The sensitivity of this assay was 9.4 pg/ml. The examination of this methodology showed that all its indices met the demands of RIA. There was no difference in the mean value of serum CT between the two sexes in 232 normal subjects, but the serum CT concentration in those aged of sixty and over of men and women, were much lower than those aged of fourty group. Mean CT concentration was low in the patients with primary osteoporosis, but were significantly higher in those suffering from medullary thyroid carcinoma, primary hyperparathyroidism crisis and chronic renal failure. PMID- 10453499 TI - [The study of mutation in exon 17 of insulin receptor gene in essential hypertensive pedigrees]. AB - Using the molecular scanning technique of single-stranded conformational polymorphism, we examined the exon 17 of the insulin receptor (INSR) gene in 44 subjects of 6 essential hypertensive pedigrees and 2 normotensive pedigrees. In addition the serum levels of glucose and insulin during an oral glucose tolerance test (OGTT); blood lipid, and plasma angiotension II and angiotensinogen were done on these pedigrees. Upon direct sequence analysis, 5 individuals were found a single nucleotide substitution at the codon 1058 (CAC-->CAT), which didn't change the amino acid sequence. Among the five individuals 4 of them were from the families with history of hypertension, only one was from normotensive pedigree. Compared with those without the mutation, the individuals with the mutation had a lower ratio of fasting blood glucose to fasting serum insulin level (P < 0.01) and an elevated plasma Ang II concentration. (There was no significant difference, P > 0.05, probably due to the mutant cases which we studied were small). Thus, we conjectured that the mutation in codon 1,058 of the INSR gene might be related with the insulin resistance in hypertensive patients and subjects with the positive hypertensive history. PMID- 10453500 TI - [Study on the inclusion compound of phyllostachysin A and DMF]. AB - The crystal structure of inclusion compound phyllostachysin A[1] and DMF, C20H26O6.C3H7ON, was determined by X-ray diffraction. A colorless block crystal with dimensions of 0.6 x 0.6 x 0.8 mm was used for analysis. It shows orthorhombic, and it's space group is P2(1)2(1)2(1) with a = 10.895(3), b = 13.237(2), c = 15.373(3) A, V = 2217.0(3) A3 Z = 4, Dc = 1.302 g cm-3. Direct method was applyed to determine the structure, and the parameters were refined by least square method. The final Rf factor is 0.060 and Rw = 0.083. The result shows that the crystal is formed by Phyllostachysin A and DMF by inclusion interaction. PMID- 10453501 TI - [Differential display analysis of human fetal brain mRNAs and isolation of brain specific novel ESTs]. AB - Using the differential display technique (DDRT-PCR) originally developed by Liang and Pardee, we tried to isolate the stage-specific or tissue-specific genes in the developing human neural system (brain). With about fifty sets of arbitrary primers and anchored primers, DDRT-PCR was carried out to analyze the differentially expressed mRNAs between human fetal brains of different developmental stage or different parts of human fetal brain at the same developmental stage. About one hundred bands containing cDNA fragments of differential interest were cut out from the polyacrylamide gel, thirty-four of which were cloned and sequenced. They were accepted as novel cDNA sequences by GeneBank. In order to check the feasibility of DDRT-PCR, three of the thirty-four cDNA sequences were randomly chosen as probes for further analysis by dot blot RNA hybridization, and two proved to be specifically expressed in human brain. PMID- 10453502 TI - [The expression of MLC2-chymase fusion gene in cultured neonatal myocardial cells]. AB - A series of MLC2-chymase fusion genes consisting of nested 5' deletion of the chymase promoter region were constructed and transfected into primary cultured neonatal Wistar rat myocardial cells according to the modified calcium phosphate precipitation method. Total RNA Dot-blot hybridization and analysis of densitometric scanning demonstrated that MLC2 promoter is enough to direct human heart chymase gene transcription in cultured neonatal rat myocardial cells. The highest level of expression was found with the MLC2-chymase construct containing 6 bp of the chymase promoter region. More sequence of the chymase promoter region between MLC2 promoter and the ATG of chymase gene resulted in a subsequent decrease in the expression of human heart chymase gene. PMID- 10453503 TI - [Immunohistochemical study on the distribution and coexistence of SP, VIP and NPY in the rat submandibular gland]. AB - The distribution and coexistence of SP, VIP and NPY in the submandibur gland of rat have been studied with immunohistochemical ABC methods. The results indicated that SP, VIP and NPY-IR nerve fibers were presented as varicose and linear profiles, which mainly travelled around acini, duct and blood vessels. The ganglion cells in submandibular gland showed SP, VIP and NPY-IR all positive immunocytochemically. The findings indicated that the neuropeptides were coexisted in the same neuron body and might play the role of regulation of the secretary activity and effective blood supply. PMID- 10453504 TI - [Effect of hypoxia on distribution and activity of nitric oxide synthase in rat lung]. AB - Nitric oxide is an important cellular messenger molecule that has been implicated in a wide range of physiological and pathophysiological actions in cardiovascular, immune, and nervous systems. Nitric oxide synthase (NOS) is the sole and key enzyme responsible for the generation of nitric oxide. The effect of hypoxia-induced pulmonary hypertension on NOS in the lung is controversial. To clarify the effect of hypoxia on distribution and activity of NOS in rat lung, localization of NOS in the lungs of hypoxic and normoxic rats were studied using NADPH-diaphorase histochemical staining techniques, NOS enzyme activity in the lung homogenates was assessed by [3H] arginine to [3H] citrulline conversion. In the normoxic rat, NADPH-d was distributed in the endothelial cells of large pulmonary vessels (ID > 150 microns) and medium-sized (50 microns < ID < 150 microns) vessels but was not detected in the endothelium of small vessels (ID < 50 microns), and there was an absence of NADPH-d staining in the smooth muscle cells of small, medium, and large pulmonary vessels. However, after hypoxic exposure for two weeks, NADPH-d staining increased dramatically in the endothelial cells of large and medium-sized pulmonary vessels, and NADPH-d became markedly positive in the endothelial cells of small vessels. Hypoxia was also found to induce de novo NOS expression in the smooth muscle cells of small, medium-sized, and large pulmonary vessels. The enzyme activity of constitutive NOS(cNOS) was decreased obviously in the hypoxic rat lungs, but that of inducible NOS(iNOS) was increased significantly in the hypoxic rat lungs. These results suggested that the inhibited endothelium-derived relaxing factor (EDRF)/NO dependent vasodilation after hypoxic exposure might be induced by decreased activity of cNOS in the endothelium of pulmonary vessels, and hypoxia-induced upregulation of iNOS expression and activity in the rat lung might play an important role in the adaptation of pulmonary circulation to hypoxia. PMID- 10453505 TI - [Contents of inositol phosphates and response to phenylephrine in aorta from both young and old rats]. AB - The changes of contents of inositol phosphates in Phenylephrine stimulated contraction of aorta from both 3 month and 20 month Wistar rats were studied by means of Bioassay method, and [3H]-myo-inositol labelling, ion exchange chromatography. The results were as follows: (1) In both resting and PE-activated arteries, IP took the greatest part (67%-83%) in total amount of inositol phosphates and the contents of IP, IP2 and IP3 were significantly higher in 20 month rat aorta than that in 3 month rat. (2) There is an age-related reduction in aortic contraction in Ca(2+)-free buffer and Ca(2+)-containing buffer. These results suggest that there is an age-related difference in IP metabolism pattern and pathway, and that IP metabolism in old rat is obviously active, but the Vascular response is apparently low. PMID- 10453506 TI - [HIV-1 env glycoprotein gp120 and gp160 expressed in vaccinia virus system and their antigenicity analysis]. AB - Human immunodeficiency virus type 1 (HIV-1) is the principal causative agent of AIDS that is an important concerns to human being health. The most important antigen of HIV-1 is its envelope glycoprotein gp160 (gp120 + gp41). By means of genetic engineering techniques, HIV-1 gp120 and gp160 were constructed and expressed in the recombinant vaccinia virus system under the control of bacteriophage T7 promoter or vaccinia virus p7.5 promoter. The results showed that the recombinant gp120 and gp160 expressed in this system exist in the form as glycoproteins, resuming the native epitope conformation of the viruses from which they were derived. These recombinant gp120 and gp160 could be recognized by the virus-specific neutralizing monoclonal antibodies. The findings suggested that the recombinant HIV-1 gp120 and gp160 could be used as a candidate recombinant protein vaccines. PMID- 10453507 TI - [The relationship of the permeability coefficients of testosterone with solubility parameters of vehicles]. AB - The effects of vehicles on the penetration through rat skin of testosterone were investigated. The solubilities of testosterone in various vehicles and the fluxes through rat abdomen skin had been determined. The solubility parameters of testosterone (delta i) and vehicles (delta v(n)) were calculated by Hildebrand's method. There was a good inverse parabola correlation between the log permeability coefficients (logp) and (delta v(n)). When delta v(n) approched delta i the permeability coefficients of testosterone approched the minimum as its limit. PMID- 10453508 TI - [Painful mild scoliosis in the elderly]. AB - We followed up and analysised 25 patients with slight painful scoliosis including symptoms, signs, images, as well as treatment and effects. The results showed that the mild scoliosis with rotation in lumbar and thoracic-lumbar region will develop and aggraviate with age. In the elderly, it may cause severe low-back pain intermittent claudication and neurological defect of lower extremities due to stenosis and spondylolethesis. We think this kind of patients should be treated with the anterior Zielke operation in early stage in order to correct rotation and prevent the later stage problems. If the symptoms and signs as described above have occured, surgical decompression and internal fixation should be recommended, the operative treatment is more effective than the non-operative treatment. PMID- 10453510 TI - [Effects of the double chamber intra-aortic balloon pumping on the renal circulation]. AB - The effects of the double chamber intra-aortic balloon (DIAB) and the single chamber intra-aortic balloon (SIAB) on the hemodynamic parameters in the kidney were studied. No negative effect of both cases was observed. Before and after removing the renal nerve system, the DIAB was used and the results had been compared. The pulse produced by pumping activized the renal nerve system and reduced the renal vascular resistance. Therefore the reduction of the renal vascular resistance is an important reason for increasing the renal blood flow during intra-aortic balloon pumping. PMID- 10453509 TI - [Cardiovascular effects of diabetic autonomic neuropathy in NIDDM with coronary heart disease]. AB - 42 coronary heart disease (CHD) (21 with NIDDM) were studied with a noninvasive cardiovascular autonomic function tests Namely; 1. Heart rate response to valsalva manoeuvre; 2. Immediate HR response to standing; 3. BP response to standing; 4. BP response to sustained handgrip, on two groups; Group 1: 21 NIDDM with CHD; Group 2: 21 CHD. The results showed that the incidences of abnormal of 4 tests were: 28.6%, 42.9%, 19.0%, and 42.9% in Group 1 respectively, and all were 0% in Group 2. Significant difference (P < 0.05) was found between two groups. It suggested that cardiovascular autonomic function tests is useful in diagnosing autonomic neuropathy in NIDDM with CHD patients. PMID- 10453511 TI - [Experimental studies on somatic gene therapy for diabetes. I. Structuring of recombinant from human insulin gene and ammalian expression vector PRC/CMV]. AB - To develop a model somatic gene therapy system for diabetes, we constructed a human insulin expression vector in non B cells. As the first step, an insulin cDNA fragment of 260 bp, generated by a complet digestion of PBCA with EcoR I and BamH I, was inserted into the EcoR I/BamH I site of plasmid PBS.SK by ligation of cohevise-ended DNA to construct transition plasmid PBS.INS. Then the plasmid PBS.INS was completly digested by Hind III and Xbal I. The small DNA fragment containing insulin cDNA gene was subcloned to the expression plasmid PRC/CMV to form recombinant PRC/CMV.INS. PMID- 10453512 TI - [Amplification and sequencing of heavy chain variable region (VH) gene of a human monoclonal antibody (CM-1) with specificity to breast cancer]. AB - For rescueing the heavy chain variable region (VH) gene of CM-1--a human monoclonal antibody against breast cancer, total RNA was extracted from the CM-1 hybridoma cells. After preparing its cDNA by reverse transcription and amplifying by PCR, a DNA fragment with about 400 bp was obtained. With an internal primer, the fragment was directly sequenced and 344 bases which consisted well to known human antibody VH gene construction were read out. In addition, its corresponding amino acid sequences coincided with the 1-3 group canonical structure of human antibody variable region. So we suggested that CM-1 could be considered as an individual of human antibody family, and should aid the preparation of antibody molecules. PMID- 10453513 TI - [Antitumor effect on melanoma by in vivo direct injection of packaging cells transfected recombinant IL-2 gene]. AB - Our previous studies showed that ex vivo transfer of human IL-2 gene into murine B16 melanoma cells resulted in the expression of IL-2 activity and had inhibitory effect on tumor growth in vivo. In this study, B16 melanoma bearing mice were injected directly with IL-2 recombinant PA317 cells. The in vivo gene transfer revealed significant reduction of tumor size and prolongation of survival of tumor bearing mice. Using slot blot analysis, the expression of IL-2 mRNA in tumor mass was detected and the IFN gamma activity was also enhanced in gene transferred mice. These results indicated that in vivo direct transfer of IL-2 recombinant retrovirual vector producing cells would induce transfer gene into tumor cells and exert its antitumor effect. PMID- 10453514 TI - [Changes in t-PA, PAI-1 gene expression of coronary arteries with preconditioning in acute myocardial infarction rat]. AB - The changes of t-PA, PAI-1 gene expression and t-PA activity were investigated in coronary arteries of Acute Myocardial Infarction (AMI) rat by nucleic acid molecular hybridization and spectrophotometric assay. The effect of preconditioning on t-PA and PAI-1 was observed in this model. The results indicated that: (1) Tissue t-PA activity in ischemia was significantly decreased than that in control (P < 0.01). t-PA activity was obviously increased in preconditioning group (P < 0.01 compared with ischemia group). (2) t-PA and PAI-1 mRNA expression in ischemia were higher than that in control, especially PAI-1 4.6 times higher than that in control), while PAI-1 expression lower in preconditioning rat. (3) In situ hybridization showed t-PA and PAI-1 were primarily expressed by endothelial cells in vessels of rat heart. These data suggested that abnormal expression of PAI-1 and t-PA was involved in AMI. This fibrinolytic depress was associated with high PAI-1 expression. Preconditioning may be effective to protect myocardium through decreasing PAI-1 mRNA expression. PMID- 10453515 TI - [Beneficial effect and mechanism of 764-3 on cognitive deficit induced by brain damage in rats]. AB - Animal model of excitotoxic cognitive deficit induced by kainic acid bilaterally injection into caudate nucleus was successfully established in rats. Based on this model, chronic administration of 764-3 (8 mg/(kg.d), i.p.), an extract from Chinese traditional medicine, markedly improved the learning and memory impairment in the injured caudate nucleus rats. Moreover, attenuation of the neuronal loss in caudate nucleus following excitotoxic damage was observed after 764-3 treatment. In addition, results indicated that the mechanism of the effect of 764-3 may be related to the antagonism against the decline of Na(+)-K(+) ATPase and Ca(2+)-Mg(2+)-ATPase activity in synaptic membrane resulted from excitotoxicity. PMID- 10453516 TI - [Studies on all-spectrum analysis for X-ray diffraction of Chinese herbal medicine calculus bovis]. AB - Investigation on famouse Chinese herbal medicine-Niu huang (calculus bovis) was carried out by all-spectrum X-ray diffraction analysis. Diffraction spectrums, as well as the specific symboling peaks of calculus bovis, artificial bezoar, bile ductstone, human gallstone and hog gallstone, were recognized. The error distribution curves of d-delta d for specific symboling peaks was also obtained by calculation under diffrent testing conditions, by which we identified successfully three samples provided by a pharmaceutical factory. This article shows that all-spectrum X-ray diffraction analysis can be used to identify Chinese traditional crude drug, and provides for morphological and microscopical study. PMID- 10453517 TI - [Changes of 5-HT2A receptor binding on platelet of patients with CAD and the effects of exercise test]. AB - It is known both 5-HT2A receptors of platelet and vascular smooth muscle regulate vascular constriction and platelet aggregation to 5-HT challenge. And that 5-HT induced hypperreactivity of vascular smooth muscle and platelet were noted in patients with coronary artery disease (CAD). In order to investigate the mechanism of platelet hyperreactivity in cardiovascular disease. Platelet 5-HT2A receptor were measured by specific binding of 3H-ketanserin in patients with CAD (CAD group) and patients with chest pain but without CAD as well as healthy subjects (control group) before and after submaximal treadmill exercise. At the same time the levels of plasma 5-HT were also measured. The results showed: (1) In the resting state, 3H-ketanserin exhibited saturable binding with Bmax of 71.1 +/- 10.1 fmol/mg protein, Kd 1.35 +/- 0.20 nmol/L in control group (n = 8) and with Bmax of 102.1 +/- 22.2 fmol/mg protein, Kd 2.44 +/- 0.66 nmol/L in CAD group (n = 10). Between two groups had no significant difference (P > 0.05). And the specific binding of 2.0 nmol/L 3H-ketanserin and the level of plasma 5-HT in CAD group were higher than those in control group (P < 0.05). (2) After exercise all of the specific binding of 2.0 nmol/L 3H-ketanserin, Bmax of receptor binding and the level of plasma 5-HT were significantly increased in CAD group but not in control group. The above results suggested that the up-regulation of platelet 5 HT2A receptor was involved in the platelet hyperreactivity in CAD. The submaximal treadmill exercise had no significant effect on platelet function in control group, but it could lead to platelet 5-HT2A receptor increment and platelet activation in CAD group. Therefore, these changes of platelet might play a role in the acute myocardial ischemia induced by exercise. PMID- 10453518 TI - [The cloning, expression, purification and bioactivity identification of recombinant IGF-I]. AB - The fragment containing IGF-I cDNA and cyc1 terminator from PSK-IGF-I plasmid was cloned into PSK43 SB treated by Hind III, Klenow and Cla I, generating a recombinant PSB-IGF-I (DH), which have double Hind III sites. This plasmid was treated by Hind III and Klenow, ligated to give a recombinant plasmid PSB-IGF I(NH), which has no Hind III site. After analysed by restriction endonucleases and sequencing, the fusion site was verified to be correct. This plasmid was excised and a BamH I-Cla I fragment containing yeast alpha-factor promotor, leader sequence, IGF-I cDNA and cycl terminator was cloned into yeast episomal plasmid vector YEpHC8, generating a recombinant plasmid YEpHC8-IGF-I. Transformed this plasmid into yeast competence cells BJ1990 by using LiAc method. The expressed products were secrected into the medium broth and having the correct molecular weight of 8,100 on SDS-PAGE. The products of IGF-I were raised by using high-cell-density fermentation of Saccharomyces cerevisiae. The cleared supernatant of yeast medium was applied to Bio-Rex 70 resin, eluted with 1 mol/L ammonium acetate, pH8.0. Main peak was pooled and applied to Bio-Gel P10 (200-400 mesh). The second peak was collected and get relatively pure IGF-I proteins. The biological activities of IGF I product was assayed in NIH3T3 cells by using MTT method. The results show that the expressed IGF-I can obviously stimulate NIH3T3 cells to proliferate at the concentration ranging from 10 ng/ml to 50 ng/ml, suggesting that the protein has its biological activities. PMID- 10453519 TI - [Sequence of HLA-DQA1 promoter region in the Han people]. AB - Polymorphism of HLA-DQA1 promoter region (QAP) in the Han people has been identified. The results revealed a number of differences, some of which are in the critical class II boxes, and generally conserved in HLA-DQA1 promoter region. The major differences occurred in the X box, Y box and S box. Within the X box, the Hans carry a A at position -111, instead of a G, and a G or a A can be present at position -98. Within the S box, the Hans carry a G at position -131. Within the Y box, position -71 is a A rather than a G. Some single base substitutions have been detected from IDDM patients at the 5'-flanking region of the S box and between X box and Y box. Particularly, the insertion of CCA bases has been identified at the position between -157 and -158 in a IDDM patient. These data suggest that the polymorphism of HLA-DQA1 promoter region may play a role in susceptibility to IDDM. PMID- 10453521 TI - [A deep regional impedance method measuring the blood distribution in the lung]. AB - A new deep regional impedance method to measure the blood distribution in the lung has been developed. Using a multi-electrode system, the method simultaneously detects impedance information of chest, and back including lung and the thorax, then extracts the impedance information directly from lung with a special data processing approach in order to eliminate thorax effect on the measurement. The salt pool experiment and the preliminary application of measuring the blood distribution in the lung have been made. The results show that the method raises the relative sensitivity and the distinguishability of the blood measurement in the lung, and as a new deep regional impedance measurement method, it has a good prospects of clinical application in the future. PMID- 10453520 TI - [beta-Endorphin enhances IL-2 and IFN-gamma gene expression in human blood mononuclear cells]. AB - The effect of beta-endorphin (beta-END) stimulation of cytokines production by PHA-induced human peripheral blood mononuclear cells (PBMC) was studied by reverse transcription in combination with polymerase chain reaction (RT/PCR), slot blotting and hybridization to detect and measure messenger RNA (mRNA). beta END significantly and dose dependently enhanced IFN-gamma and IL-2 mRNA expression of T cells and naloxone (Nal) reversed the effect of beta-END on IL-2 mRNA expression. The results further demonstrate the important link between the neural system and the immune system. PMID- 10453522 TI - [Construction of poliovirus cDNA chimera with (NKNDD)n and (NANP)n]. AB - Oligonucleotides encoding one copy of NKNDD, containing a P.f. B epitope NKND, and two copies of CSP repeated B epitope NANP were synthesized. After self ligation, NKNDD and (NANP)2 were cloned into the intermediate vector pSKMM. Series of clones with different copies of NKNDD and NANP were identified. (NKNDD)n/MM with three, four, five, eight copies of NKNDD and (NANP)n/MM with four, six copies of NANP were chosen. Then (NKNDD)n and (NANP)n of these recombinants were recovered respectively and finally inserted into the N-Ag I site of poliovirus type I Mahoney strain full-length cDNA, which is contained in the expression plasmid pSV202H+. As using semi-quantitative hybridization in identifying tranformants, the procedure in our experiment for cloning different copies of NKNDD and NANP was greatly simplified. PMID- 10453523 TI - [Evaluation of emergency surgery in gestational trophoblastic tumours]. AB - We retrospectively analyzed 27 cases of gestational trophoblastic neoplasia treated by emergency surgery from 1985-1996 at PUMC hospital. Seventeen cases were diagnosed of choriocarcinoma and 10 were invasive mole. Sixteen out of 27 patients were subjected to hysterectomy because of uterine perforation or severe uterine bleeding; 4 cases received unilateral oophorectomy because of torsion of theca lutein cyst. Emergency open surgery and excision of the metastatic brain tumour were undertaken in 3 patients with elevated intracranial pressure caused by brain edema and haemorrhage; 2 patients showed lifethreatening haemorrhage from vaginal metastatic tumour and were managed by operative intervention. Partial jejunectomy was performed in 2 patients due to rupture of jejunal metastatic mass. Of these 27 cases, 17 hadn't been treated with chemical reagents and 6 cases had received one course of chemotherapy before surgical procedures. After multiple courses of combined chemotherapy postoperatively, 23 patients had achieved complete remission. It is concluded that surgical intervention plays an important role in patients with trophoblastic disease when emergency situations (e.g. life-threatening haemorrhages) occur; early diagnosis and prompt initiation of chemotherapy might have rendered surgery unnecessary. PMID- 10453524 TI - [Changes in airway space following mandibular setback using sagittal split ramus osteotomy with rigid internal fixation]. AB - The changes in airway space following mandibular setback using sagittal split ramus osteotomy and rigid internal fixation were studied in 28 Japanese patients with mandibular prognathism. Lateral cephalometric radiographs were taken immediately before and shortly after surgery as well as 5 to 12 months postoperation. The mean setback of mandible was 8.3 mm in the right side, and 8.2 mm in the left side. The mean amount of relapse of pogonion point during the following-up period was 0.52 mm horizontally, and 0.92 mm vertically. A significant decrease in airway space width and area, espacially in the lower part of airway space was found following mandibular setback shortly after surgery. Although there was some increase both in airway space width and area during the following-up period, they did not increase to their original values. This suggests that the narrowing of airway space following mandibular setback using sagittal split osteotomy can be permanent. No significant correlation was found between the amount of mandibular setback and airway space changes, and no significant correlation was found between the amount of airway space changes and relapse of pogonion point during the following-up period. PMID- 10453525 TI - [Spinal segment distribution of neural innervation related houhai acupoint- studied by CB-HRP tracing method focused on observation of the dendrites of spinal motor neurons]. AB - The spinal segment distribution of neural innervation related Houhai acupoint in rats has been investigted by using horseradish peroxidase conjugated cholera toxin B subunit (CB-HRP) tracing technique. Spinal motoneurons (Mn) in L6-S1 were labeled retrogradely by CB-HRP, and their features have been characterized: 1) their marginal plexus of the dendrites radially projected to the lateral and/or anterior funiculus and their distal portions forming subpial dendritic plexus; 2) the medial dendrites ran along the ventral margin of the gray commissure and their distal portions surrounded the central canal' forming a subependymal dendritic plexus. Both of them are Golgi-phobic. Some distal branches of dendrites penetrated to the contralateral gray matter. The dendritic arbors of the labeled motoneurons in the group of electro-acupuncture or group of acupoint inoculation with sheep red blood cells contracted in comparison with that in the normal control group. The primary sensory transganglionic labeled fibers and preterminal fibers revealed by CB-HRP in the dorsal horn and dorsal commissural nucleus of the spinal cord (L5-S2) converged and overlaped with the dendrites of retrogradely labeled motoneurons. The results indicated there is a reflect circuit of afferent-integration-efferent neuro-network in the spinal cord related Houhai acupoint. The Mns retrogradely labeled by CB-HRP in the L6-S1 segments might be involved in Houhai acupuncture. PMID- 10453526 TI - [Preliminary application of mRNA differential display technique in the isolation of genes possibly associated with tumor metastasis]. AB - We have previously reported that in vitro treatment with interferons of MA891, a spontaneous mouse mammary tumor capable of metastasizing to the lungs upon subcutaneous inoculation, could modulate its metastatic potential. IFN-gamma increases while IFN-alpha decreases lung metastasis. Identifying the genes differentially expressed in MA891 cells separately treated with IFN-alpha and IFN gamma. MA891 cells were treated in vitro with 200 U/ml IFN-alpha or IFN-gamma respectively for 48 h. Total RNA was isolated with the single step method of acid guanidinium thiocyanate. After being digested by DNase I, the two RNA samples were differentially displayed with T12MA/T/C/G x 20 arbitrary primer pairs. Most of the RT-PCR were done twice. The differentially expressed cDNA bands were isolated, reampliated and cloned. Ten differentially expressed cDNA bands were identified: T2 alpha, T2 gamma, C4 alpha, T6 alpha, T11 gamma, G15 gamma, T16 gamma, T5 alpha 1, T5 alpha 2, T17 alpha, T15 alpha, T15 gamma, named according to the type of IFN and primer pairs used. The total RNA samples of MA891 cells treated by IFN-alpha and IFN-gamma were differentially displayed with T12MA/T/C/G x 20 arbitrary primer pairs. Theoretically they can cover 95% mRNAs. It is anticipated that one or more of the ten cDNA clones may be related to the metastatic potential of MA891 cells. PMID- 10453527 TI - [Erythropoietin gene cloning and expression in S. cerevisiae]. AB - Mature human erythropoietin gene was amplified from EPO cDNA by PCR methods. The PCR product was cloned into pUC18 plasmid at Sma I site, then precisely engineered into a intermidiate vector pSK43SB which were digested with Hind III, Mung bean nuclease, and Sal I. Then degest pSK43SB-EPO plasmid with EcoR I and Cla I, the EC fragment with an alpha-factor leading sequence, EPO gene and CYC1 terminater were produced. It was then cloned into a typical high efficiency episomal expression vector YEpHC8. Human EPO protein with highly mannose glycosylated was identified by Western blot methods in both secreted and in cells proteins. N-Glycosidase F digested secreted EPO can produce 20,000 EPO without N glycosylation similar with that produced in cells. PMID- 10453528 TI - [Study on small hepatocellular carcinoma: a review of 20 years experience]. PMID- 10453529 TI - [Effect of pathogen-stimulated human CD4+ T cells on gamma delta T cells]. AB - OBJECTIVE: CD4+ helper T lymphocytes and gamma delta T cells play important role in antimicrobial immunity, however, the mechanism of interactions in question still remains to be elucidated. METHODS: CD4+ T cells and gamma delta T cells separated from peripheral blood lymphocytes of healthy donors by panning technique were stimulated with intracellular pathogen-Mycobacterium tuberculosis and human herpes virus-6 as well as extracellular pathogen-Trichomonas Vaginalis respectively in vitro. RESULTS: The stimuli induced CD4+ T cells significantly to proliferate and producing Th1-type cytokine profile-IL-2 and IFN-gamma. With a semipermeable co-culture system in vitro, antigen-activated CD4+ Th1-like cells were found to promote the proliferation and cytotoxicity of gamma delta T cells via soluble factors. CONCLUSION: Functions of gamma delta T cells might be inhanced by CD4+ cells with Th1-type cytokines in antimicrobial immunity. PMID- 10453530 TI - [Study on the effect of continuous administration of IL-1ra in BXSB mice]. AB - OBJECTIVE: To explore the consequences of IL-1 blocking in BXSB mice which is an experimental model for human SLE. METHODS: rh IL-1ra was expressed in E. coli and injected in BXSB mice. 13 of 4.5 month-age male BXSB mice were divided into two groups, one group was injected rh IL-1ra 10 times (twice a week) at dose of 400 micrograms per mouse each time, another group was injected PBS at the same time as control. We monitored serum ANA and proteinuria weekly, and detected IgG, C3 deposition and IL-6 expression in kidneys at 40th day. RESULTS: The results showed that the increased level of serum ANA and proteinuria in treatment group were not higher than in control group, the IgG, C3 deposition and IL-6 expression in kidneys and IL-6 activity in serum of the treaed group were lower than the control group, whereas no difference of GPT level in the two groups. CONCLUSION: IL-1 might play a pathogenic role in BXSB mice. Blocking or reducing IL-1 secretion would be of beneficial to the treatment of SLE. PMID- 10453531 TI - [Effects of PGMS on the functions of human platelets and endothelial cells]. AB - OBJECTIVE: To study the effects of Propylene glycol mannte sulfate (PGMS), a heparin-like substance on the functions of platelets and endothelial cells. METHODS: Using 125I labeled monoclonal antibody to measure the expression of GMP 140 on the surface of human platelets; and using naphthol blue black stain method to study the growth of human umbilical vein endothelial cells (HUVEC) and human umbilical vein smooth muscle cells (HUVSMG). RESULTS: PGMS significantly inhibited thrombin stimulated expression of GMP-140 on the surface of human platelets. IC50 was 127 micrograms/ml. Indomethacin also inhibited GMP-140 expression, but its effect was weaker than that of PGMS with the IC50 of 502 micrograms/ml. PGMS did not affect the growth of HUVSMC. On the contrary, it significantly promoted the growth of HUVEC at very low concentration (1.6 micrograms/ml). CONCLUSIONS: PGMS showes anticoagulative effects on both platelets and endothelial cells and it might well be developed as a new antithrombotic and anticoagulateve agent. PMID- 10453532 TI - [Study of mechanism of PTH modulating cytosolic free calcium in rat DCT]. AB - OBJECTIVE AND METHODS: In order to understand the mechanism of PTH in regulating cytosolic free calcium concentration ([Ca2+]i), we differentiated rat renal proximal and distal convoluted tubules (PCT&DCT) by Percoll density gradient centrifugation, and successfully gauged the effect of hPTH (1-84), dibutyryl cAMP (db cAMP, a kind of cAMP activator) and prostaglandin E2 (PGE2) on [Ca2+]i of DCT using fluorescent Ca2+ indicator Fluo 3-AM. We found that PTH, db cAMP and PGE2 all could increase [Ca2+]i. RESULTS: The results suggest that the rise in [Ca2+]i induced by PTH in DCT was through activation of cAMP, and PGE2 might be also involved in regulation of [Ca2+]i as a messenger of PTH. PMID- 10453533 TI - [The effects of E2 and it's metabolites on the proliferation of rat anterior pituitary cells in vitro]. AB - OBJECTIVE AND METHODS: The present work is determined to observe the effects of E2 (estradiol), 2-OHE1 (2-hydroxyestrone), 2-OHE2 (2-hydroxyestrodiol) on the proliferation of rat anterior pituitary cells (APC) in vitro by laser scanning confocal microscopy. RESULTS: 10(-6) mol/L E2 stimulated the growth of APC. After 2 days of incubation with E2, the DNA content of APC increased to 1.3 times of the control group (P < 0.01). 10(-6) mol/L 2-OHE2 (other than 2-OHE1) stimulated proliferative activity of APC and inhibited the inhibitory effect of peribidil (10(-5) mol/L), a dopamine receptor agonist, on the poliferative activity of rat APC. PMID- 10453534 TI - [Melatonin inhibits TRH-stimulating prolactin gene expression of anterior pituitary cells in newborn rat in vitro]. AB - OBJECTIVE AND METHODS: This work was to investigate whether melatonin (MEL) plays a role in the gene expression of prolactin (PRL), by the Method of in situ hybridyzation. RESULTS: Our results indicated that, at a higher concentration, MEL not only inhibits TRH (thyrotropin releasing hormone) stimulating PRL gene expression of anterior pituitary cell in newborn rat, but also exerts a direct inhibitory effect on PRL gene expression in vitro. CONCLUSION: These results suggest that MEL may be a regulator of PRL synthesis and secreting in vivo. PMID- 10453535 TI - [Guided tissue regeneration around dental implants in immediate extraction sockets: comparison of resorbable and nonresorbable membranes]. AB - OBJECTIVE: This study was performed to compare the efficacy of guided tissue regeneration (GTR) around dental implants immediately placed into extraction sockets by resorbable or nonresorbable membranes. METHODS: Mandibular P2, P3, and P4 of four adult beagle dogs were extracted bilaterally, and buccal standard defects were created and measured. Eighteen commercially pure titanium Steri-Oss implant fixtures were placed into the fresh extraction sockets. Four untreated implants were as controls, four implants received polytetrafluoroethylene (e PTFE, Gore-Tex) membranes, five implants received collagen membranes, and five implants received polyglactin 910 mesh. After 14 weeks, clinical measurements were taken and the dogs were sacrificed and all specimens retrieved for histologic and histomophometric evaluation. RESULTS: The average gain in bone height was 2.1 mm for untreated control sites, 3.3 mm for Gore-Tex sites, 3.8 mm for collagen sites, and 1.3 mm for polyglactin 910 sites. CONCLUSION: The results of this study indicated that Gore-Tex and collagen membrane produced good results for GTR around implants. Collagen membrane can be a valid alternative to Gore-Tex membrane to improve bone regeneration around dental implants, while polyglactin 910 mesh seemed not suitable to be used as GTR membrane in immediate implantation for its high infection rate. PMID- 10453536 TI - [Relationship between the activity of leukocyte chemokines in gingival crevicular fluid and periodontitis]. AB - OBJECTIVE AND METHODS: Chemotactic activity in gingival crevicular fluids (GCF) from 39 cases of patients with periodontitis and 10 cases of healthy donors was determined via Boyden method. RESULTS: Chemotactic activity in GCF from patients was much higher than that from healthy donors. Chemotactic activity in GCF was not relative to pocket depth and attachment loss, but markely correlative to gingival index and bleeding index. CONCLUSION: The results indicates that leukocyte chemokine activity would not reflect the degree of periodontal damage, but might be a marker for periodontal inflammatory reaction and disease activity. It is likely that interleukin-8 might be a major component among the chemokines in GCF. PMID- 10453537 TI - [Evaluation of parotid sialography and labial salivary gland biopsy to the diagnosis of Sjogren syndrome]. AB - OBJECTIVE: To investigate the relationship between the change of parotid sialograms and the results of labial gland biopsies in the patients with Sjogren syndrome (SS). METHODS: Parotid sialography and labial salivary gland biopsy were performed on 100 patients with Sjogren syndrome. RESULTS: The sialectasis of parotid terminal ducts was found in 87 patients, the widening only of main ducts was found in 8 patients, and 5 cases showed normal sialograms. The results of labial salivary gland biopsies showed, that 90 cases had focal lymphocytic sialadenitis (Zhen Linfan's grade score of 2), 59 cases had a score of 4 accoding to Chisholm's grading score; moderate scattered lymphocitic infiltration was shown in 10 patients. CONCLUSION: Our results suggested that the lesions of parotid gland and labial salivary gland in SS were concerted closely. Therefore a new way of combining the changes of parotid gland and labial salivary gland for the diagnose of SS was proposed. PMID- 10453538 TI - [The inhibitory effects of antioxidant vitamins on serum oxLDL and experimental atherosclerosis of rabbits]. AB - OBJECTIVE AND METHODS: In order to study the inhibitory effects of antioxidant vitamins on serum (low oxidative density lipoproteins, oxLDL) and experimental atherosclerosis in rabbits, 20 rabbits were fed on cholesterol rich diet and antioxidant vitamins (vitamin E, vitamin C and beta carotene) for 12 weeks. oxLDL were tested by ELISA at the beginning of experiment and after 4 weeks 8 weeks. RESULTS: The results showed that supplement of antioxidant vitamins can decrease the oxLDL level significantly and inhibited development of atherosclerosis lesion around aorta in rabbits. PMID- 10453540 TI - [Drilling decompression of the proximal tibia for osteoarthritis of the knee]. AB - OBJECTIVE: To investigate the effect of drilling decompression of the proximal tibia for the Osteoarthritis of the Knee. METHOD: The drilling decompression of the proximal tibia were performed in 21 patients (38 knees) of osteoarthritis with 99mTc-MDP scintingraphy and the intraosseous phlebography made pre postoperatively. Knee function scores and scales were evaluated. RESULT: During 6 12 months follow-up, the preoperative score of the knee function was 18-48 (average 30.78) and the postoperative score was 61-96 (average 87.95); the rates of fair and good of knee scales rose from 5% preoperatively to 86.84% postoperatively. CONCLUSION: Drilling decompression of the proximal tibia is a simple, safe, less traumatic and effective method to relieve the pain and improve the function for the early osteroarthritis of knee, For late cases it is a method to relieve pain. 99mTc-MDP scintingraphy can be used as a method to diagnose the early stage of the osteoarthritis. PMID- 10453539 TI - [Effect of acute moderate isovolumic hemodilution on the pharmacokinetics of vecuronium]. AB - OBJECTIVE: To determine the effect of acute moderate isovolemic hemodilution on pharmacokinetics of vecuronium. METHODS: Twenty-six adult patients, ASA grade I, undergoing elective plastic surgery, were randomly divided equally into the control and hemodilution group. The blood samples were taken for 5 hours following intravenous bolus of 100 micrograms/kg of vecuronium in two groups. The plasma concentrations of vecuronium were determined with an improved fluorometry and the pharmacokinetic parameters were obtained by fitting the data with a 3P87 program. RESULTS: The disposition of vecuronium can be best described by a three compartment open model. As comparison with the control group, the values of Vc and Vdss in hemodilution group were greater than that in the control, while T1/2 beta was markedly prolonged in this group. There were no significant differences in T1/2 phi, T1/2 alpha, K12, K21, K13, K31, K10, MRT, AUC, and Cl between the two groups. CONCLUSION: Acute moderate isovolemic hemodilution could change the pharmacokinetics of vecurinium, particularly in distribution and elimination phases. PMID- 10453541 TI - [Effects of calcium channel blockers and calmodulin inhibitors on the secretion of endothelin-1 in cultured endothelial cells]. AB - OBJECTIVE: The study is to investigate the effects of calcium channel blockers and calmodulin inhibitors on the secretion of endothelin-1 (ET-1) in cultured endothelial cells from human umbilical veins. RESULTS: Results showed that calcium channel blockers verapamil (5.5 x 10(-6) mol/L, 5.5 x 10(-5) mol/L), diltiazen (2.4 x 10(-4) mol/L) and calmodulin inhibitors chlorpromazine (3.1 x 10(-5) mol/L, 3.1 x 10(-4) mol/L), berbamine (1.6 x 10(-5) mol/L, 1.6 x 10(-4) mol/L) significantly decreased medium ET-1 levels in cultured endothelial cells. CONCLUSION: It was also indicated that extracellular calcium influx and calmodulin activity were necessary to the secretion of ET-1. In addition, nitroglycerine (2.2 x 10(-3) mol/L) remarkably reduced medium ET-1 levels in cultured endothelial cells, which suggested that nitric oxide might inhibit the secretion of ET-1. PMID- 10453543 TI - [Observation on apoptosis during bone growth in mice and osteogenesis in human embryo]. AB - This paper describes cell apoptosis observed in femur of adult mice and limb bud of human embryos. We observed apoptotic cells characterized by chromatin condensation, margination and apoptotic bodies. Therefore it seems that apoptosis occurs in bone as in other organs. PMID- 10453542 TI - [Changes of PDGF-alpha and beta receptor gene expression in hypoxic rat pulmonary vessels]. AB - OBJECTIVE: To reveal the role of platelet-derived growth factor-alpha and beta receptor in hypoxic pulmonary vascular remodeling. We investigated the changes of PDGF-alpha and beta receptor gene expression under hypoxic condition. METHODS: Hypoxic pulmonary hypertension rats model was reproduced by normobaric hypoxia. Changes of pulmonary PDGF-alpha and beta mRNA were examined by in situ hybridization. RESULTS: The PDGF-alpha receptor mRNA of the hypoxia group (3 days) declined to 86% compared with that of the control group, and shooting up to 217% of control at day 7, then sustained at 224% until day 14. (all P < 0.001). PDGF-beta receptor mRNA markedly increased at day 3 after hypoxia and maintained this increasing tendency consistently. Its top was 164% at day 14 (P < 0.001). The growth inhibitor, heparin had alleviated the increases of both rat pulmonary arterial pressure and PDGF receptor gene expression. CONCLUSION: Hypoxia led to the increase of PDGF-alpha and beta receptor gene expression in pulmonary vascular endothelial and smooth muscle cells, and might be involved in hypoxic pulmonary vascular remodeling. PMID- 10453544 TI - [Isolation of 28 new STSs at Xq27.3]. AB - A plasmid sublibrary of the 475 kb insert of YAC209G4 was constructed by using pBS II KS vector. The library of 3,500 clones having 100-600 bp inserts was screened with the probe of the 475 kb insert blocked with competitor DNA. Sixty unique single copy clones were found and sequenced. Checking with GenBank, 28 new STSs were obtained. Genbank accession numbers are U26560-26587. Three STSs were tested by PCR with appropriate primers using human genomic DNA as template and showed specific amplification bands as expected. PMID- 10453545 TI - [Transformation of rat hepatocytes in an in vitro primary culture by aflatoxin B1]. AB - Aflatoxin B1 (AFB1) is one of the major causative factors of hepatocellular carcinoma. In this study, the combined effects of AFB1 activated by human cytochrome p450 IA2 and c-myc in transformation of rat hepatocytes were investigated in an in vitro primary culture system. The expression vectors, Xm 6/c-myc was first constructed and their expression possibilities were examined in Alexander cells by immunocytochemistry. Then both c-myc and human cytochrome p450 IA2 expression vectors were sequentially transfected into newborn rat liver cells in serum-free primary culture. Results showed that p450 IA2 could activate AFB1 at concentrations as low as 5 ng/ml, and the activated AFB1 coupled with exogenous c-myc could induce rat hepatocytes to survive and grow beyond two-month limit in primary culture. During long-term in vitro culturing including four month in crisis, one of the randomly selected transformed hepatocytes with the growth advantage became immortalized. Immunocytochemical assays for CK-18 and rat albumin plus observed electron microscopic features clearly confirmed these cells derived from epithelial hepatocytes. Further characterization showed that the process of immortalization was associated with chromosomal abnormalities and elevated expression of TGF alpha. PMID- 10453546 TI - [The expression of rat brain constitutive nitric oxide synthase in NG108-15 cell]. AB - Full length cDNA of rat brain cNOS was inserted into the polylinker area of pRC/CMV with specific orientation and an eukaryocyte expression vector pCMVcNOS was obtained. The existence of cNOS gene was demonstrated by PCR amplification, using pCMVcNOS gene as the model and primers designed in accord with the internal sequence of cNOS gene. The insertion and orientation of pCMVcNOS were further verified by enzymatic cleavage. NG108-15 cells were transfected with pCMVcNOS by calcium phosphate DNA coprecipitation and lipofectin transfection. G418 resistant monoclonal cells were selected with a culture medium containing 600 micrograms/ml G418. NOS activity of each clone was assayed by monitoring the conversion of 3H Arginine to 3H-Citrulline. High expression cell lines were selected through measurement of the cytosol and particulate NOS activity. Out of 42 resistant monoclonal cell lines, 3 stable high expression clones have been finally selected. The increase of expressed cytosol NOS was more obvious. The result showed that the cell lines expressing cNOS at a high level had been obtained. PMID- 10453547 TI - [The frequencies of the vitamin D receptor gene alleles in adults of Han nationality in Beijing area]. AB - Vitamin D receptor (VDR) gene is considered as one of candidate genes related to the genetic basis of bone and mineral metabolism. In order to determine VDR genotypes, we analysed the polymorphisms, or genotypes of Bsm I, Apa I, and Taq I restriction enzymes, by PCR technique in 223 unrelated healthy adults of Han nationality in Beijing area. Some of the VDR genotypes were also confirmed by Southern blotting analysis. We discovered that Chinese had high frequencies of b, a, T alleles, as high as 95%, 75%, and 95% respectively, which were different from those in Caucasians and Japanese, but similar to those in Koreans (P > 0.05). This difference gave us further insight into the different pathogeny of osteoporosis in various ethnics. PMID- 10453548 TI - [IL-2, IFN-gamma, and TNF-alpha mRNA expression in peripheral blood mononuclear cells in patients with multiple sclerosis]. AB - We determined the cytokine messenger RNA (mRNA) expression patterns of peripheral blood mononuclear cells from 17 patients with multiple sclerosis (MS) and 11 normal controls, by using a quantitative reverse transcription in combination with polymerase reaction (RT/PCR) and Southern blot hybridization method. A significantly higher levels of IL-2, IFN-gamma mRNA in patient with MS compared to normal controls and significantly higher level of expressions of IFN-gamma, TNF-alpha mRNA were detected in active MS patients compared to those with stable desease (P < 0.05). These data further support the central role of some cytokines in the pathophysiology of this autoimmune disease. PMID- 10453549 TI - [cDNA cloning of human stem cell factor and its high level expression in E. coli]. AB - The total RNA of HepG2 cell was extracted as the template by ultrocentrifuge method. The full length cDNA (0.8 kb) encoding the human stem cell factor (hSCF) was amplified by RT-PCR method. The cDNA encoding mature hSCF (0.5 kb) was sequenced and was recombined into the expression vector (PBV-220). The expression level of rhSCF in E. coli DH5 alpha was about 15% of the total protein. PMID- 10453550 TI - [Effect of calcium and angiotensin II receptor on angiotensin II-stimulated protein synthesis in neonatal myocardial cells]. AB - In this study, the role of angiotensin II (Ang II) receptor, Ca2+ in the Ang II stimulated neoeatal myocytes protein synthesis were investigated and the effect of Ang II on expression of c-fos gene was observed. The results showed that the stimulatory effect of Ang II on protein synthesis was blocked by the Ang II antagonist-[Sar1 Ile8] Ang II. When verapamil, EG-TA, dantrolene, and Fura-2/AM, were added to the medium, the 3H-leucine incorporations stimulated by Ang II were reduced by 17%, 19%, 22%, and 27%. Using calcium ionophore-A23187 did not stimulate protein synthesis, but enhanced Ang II-induced 3H-leucine incorporation. In addition, Ang II could induce expression of c-fos gene. These results indicated that the hypertrophic effect of Ang II on the myocytes was mediated by Ang II receptor and depended on increase in [Ca2+]i and may be associated with enhanced expression of c-fos. PMID- 10453551 TI - [Efficient expression of human C1-inhibitor in CHO cells by using a dicistronic expression vector]. AB - The full-length cDNA fragment encoding human C1-INH was obtained by gene recombination techniques and a stable expression plasmid was constructed by inserting the human C1-INH cDNA into an efficient dicistronic expression vector pED. This plasmid was transfected into DHFR-deficient Chinese Hamster Ovary cells (CHO-dhfr-) by Lipofectin method, and the positive CHO-dhfr+ cell clones which stably express C1-INH was generated. Expression of C1-INH mRNA was detected by Northern blot. Expression of C1-INH protein was confirmed by immunoprecipitation and Western blot analysis. The level of expressed C1-INH was estimated at 0.4 microgram/ml by specific ELISA techniques. The biological activity of recombinant C1-INH was assayed by using the inhibition of C1 estrolytic activity. PMID- 10453552 TI - [VP4 protein of simian rotavirus strain SA11 expressed by a baculovirus recombinant]. AB - The complementary DNA copies of the gene 4 were prepared by reverse transcription and polymerase chain reaction (RT-PCR) from genomic RNA of simian rotavirus strain SA11. A complete VP4 gene was been inserted into a baculovirus intermediate vector pVL-1393 which was under control of the polyhedrin promoter. The outer capsid protein VP4 which was a major neutralization antigen of rotavirus, was expressed in high yield in Spodoptera frugiperda cell line. The VP4 protein expressed was recognized with a hyperimmunized serum directed against the rotavitus by immunoblot assay. Expressed VP4 protein own about 10 percent of the total proteins. The baculovirus recombinant expressed full-length VP4 protein was used to immunize animals. A high level of neutralizing antibody directed against parentalstrain SA11 virus was developed in the animals. The anti-vp4 antibody could block visible cytopathic effect (CPE) forming of SA11 virus in MA104 cells. Immunofluorescence test and Western blot also confirmed that the antibody could recognize specific antigen of rotavirus in infectious MA104 cells. The results suggest that VP4 protein expressed in a recombinant baculovirus has antigenicity and immunogenicity as well. It may be an important component in developing rotavirus recombinant and subunit vaccine. PMID- 10453554 TI - [Effect of B16 cells transfected with IL-2 gene on mouse immunity]. AB - IL-2 was introduced into mice B16 melanoma cell line by using retrovirus infection method. Both B16 and B16-IL-2 cells were treated with Mit C beforehand and were then inoculated intraperitoneally as vaccines in mice respectively. Hanks solution was used instead of the vaccine in the control group. The data showed that the tumor incidence rate was zero in the group receiving B16-IL-2 vaccine followed by B16 inoculation. However, incidence rate was 100% in both the B16 vaccine immunization group and in the control groups. Experiments also indicated that the proliferation of splenic lymphocytes induced by MLTR, the specific cytotoxicity of CTL against B16 cells, the activities of splenic NK, LAK, and the level of IL-2 secretion in mice immunized by B16-IL-2 were much higher than those in the mice immunized only with B16 cells as well as the controls. These data indicated that secretion of IL-2 in mice promoted specific and nonspecific anti-tumor immunity of mice. The theoretic basis was provided for IL-2-secreting tumor vaccine. PMID- 10453553 TI - [Study on the association of HLA-DR4 with IDDM susceptibility in a Chinese population]. AB - HLA-DR4 and its subtypes were studied by PCP/SSP, PCR/SSOP, and PCR/RFLP methods in 81 patients with IDDM and 106 normal controls in the Han's nationality Chinese population. The results showed no difference in frequencies of DR4 between the IDDM group and the controls (21.0% vs 17.9%), but DRB1*0405, as a subtype Dw15, was significantly associated with IDDM susceptibility (58.8% vs 21.1%, RR = 2.67, P < 0.05). In a family with 2 IDDM patients, it has been found the haplotype DRB1*0405-DQB1*0302 was to be cosegregated with IDDM. It was suggested that DW15 subtype might be associated with IDDM in Chinese because of linkage disequilibrium with DQw8 (DQB1*0302). As an additional evidence, the findings above mentioned may account for the different association of different DR4 subtypes with IDDM in various ethnic groups. PMID- 10453555 TI - [Detecting HPV DNA in tissue of epidermal neoplasms]. AB - Fifty-eight cases (mean age 56.1 +/- 17.1 years) of cutaneous squamous cell carcinoma were studied, including 17 cases of Bowen's disease, 5 cases of Queyrat erythroplasia, 16 cases of Bowenoid papulosis and 20 cases of invasive cutaneous squamous cell carcinoma. All cases were confirmed by pathological findings. HPV6, 11, 16, 18 DNA were detected by multiple pair primers polymerase chain reaction in paraffin-embedded tissues. The results showed that 2(2/17) cases of Bowen's disease, 1 case of Queyrat erythroplasia and 9(9/16) cases of Bowenoid papulosis were positive for HPV16 DNA and that 20 cases of invasive cutaneous sqamous cell carcinoma were negative for HPV16 DNA. HPV16 is closely associated with Bowenoid papulosis and HPV16 might be a factor of multistep carcinogenesis in rare cases of Bowen's disease and Queyrat erythroplasia. PMID- 10453556 TI - [Fully length MDR1 cDNA transfer conferring resistance to adriamycine on sensitive cells GLC]. AB - Human lung cancer leads the mortality of cancers and the chemotherapy is often uneffective because of drug resistance. In order to study the role of mdr-1 gene in resistant lung cancer, the fully length mdr-1 cDNA was transferred into a sensitive lung cancer cell line GLC. The mdr-1 cDNA was constructed in a retroviral vector, pDORneo. The transfection of recombinant plasmid was carried out by lipofectin. Supernatant containing infective viruses derived from a G418 resistant clone of package cell PA317 was used to infect GLC cell which is sensitive to chemotherapeutic agents. After G418 and adriamycine selections, three P-glycoprotein positive clones were isolated and the integration of mdr-1 cDNA was demonstrated by PCR of genomic DNA. The relative resistance of 3 clones to adriamycine as and elevated by 5.4, 6.0 respectively 7.8 times compared with the untransfected cell and the transcription of mdr-1 gene in these transfected cells as obviously enhanced by in situ hybridization. This results suggest that the mdr-1 gene plays a role in increasing drug resistance of human lung cancer. PMID- 10453557 TI - [The value of salivary beta 2 microglobulin concentration for diagnosis of Sjogren's syndrome]. AB - In this paper we measured salivary beta 2 microglobulin (beta 2 m) concentration in 40 patients with Sjogren's syndrome, including 20 patients with systemic lupus erythematosus, 12 patients with recurrent aphthous ulceration and 16 patients with single xerostomia, and in 40 normal controls. The results showed that salivary beta 2 m concentration was more significantly elevated in Sjogren's syndrome patients, compared with that in both normals and other patients. The measurement of beta 2 m concentration in saliva may offer a simple and valuable method for diagnosis of Sjogren's syndrome. PMID- 10453558 TI - [Study on optimal conditions for the determination of glucose by measurement of oxygen consumption rate]. AB - We studied optimal conditions for the determination of oxygen consumption rate (OCR). The assay, with a working range up to 35 mmol/L, had an intra-assay coefficient of variation of 0.7%-3.52%, and the inter-assay coefficient of variation was 4.86%. In spiking experiments, the recovery range was 98.9%-101%. The assay was compared with CX3 (Beckman Co.). A linear regression analysis yielded a slope of 1.05, an inercept of 3.82 and a correlation coefficient of 0.999. The serum of turbidity above +2 had a negative interference on the assay (> 3.4%). The obvious interference of hemolysis, icterus and vitamin C to the assay were not observed. PMID- 10453559 TI - [The clinical and pathological features of the hypopharyngeal lipomas]. AB - Lipomas of the hypopharynx are uncommon mesenchymal neoplasms. The tumours have been reported to represent approximately 0.6 percent of benign tumours of the larynx. To date, there are approximately 80 cases of laryngeal lipomas reported in the literature. In this paper we report a new case of hypopharyngeal lipoma and discuss its clinical and pathological features and treatment. PMID- 10453560 TI - [Alternative splicing of the FMR1 gene in human fetal tissues]. AB - OBJECTIVE: To gain more insight into biological role of the alternative splicing of the FMR1 gene, isoforms of the FMR1 mRNA from human fetal heart, spleen, liver and kidney were analyzed. METHODS: RNAs were isolated from the tissues of an abortion fetus (approximately 6 months postconception). Alternative splicing patterns of the FMR1 mRNA were analyzed by RT-PCR and cloning strategy. RESULTS: One isoform of the FMR1 mRNA was found to be dominant in all of the four tissues. The major isoform was as same as the dominant one in the fetal cortex but was different from the major isoform in adult brain. The difference between the major isoforms in the fetal tissues and adult brain was the splicing out or retaining the peptide encoded by exon 12 and exon 17. CONCLUSION: This result suggested a developmental switch of alternative splicing of the FMR1 gene. The difference between the major isoforms in the fetal tissues and adult brain suggested the two peptides may have special roles in related developmental stages. PMID- 10453561 TI - [Down-regulation effect of HIM82 on respiratory burst of neutrophil]. AB - OBJECTIVE: The early control of respiratory burst of Neutrophil (PMN) was studied by using McAb HIM82. METHODS: PMNs were isolated from healthy volunteers. Superoxide anion, Hydrogen peroxide were measured in the presence of HIM82 FMLP. Sustance P and the NEP activities was measured in the presence of HIM82 alone. RESULTS: The amount of O2.- and H2O2 produced in experimental group (by combining with HIM82) are (44.00 +/- 8.9)% (n = 8, P < 0.01) and (65.16 +/- 15.41)% (n = 8, P < 0.01) of control group respectively. The activity of NEP inactivated by PMA (without HIM82) is (43.29 +/- 9.41)% (n = 8, P < 0.05) and the activity is (66.48 +/- 15.53)% (n = 8, P < 0.05) in PMA-inactivated group with HIM82. CONCLUSIONS: The result mentioned above showed that the HIM82 monoclonal antibody exerts obvious down regulation on respiration burst induced by PMA and antagniation on inactivity of NEP. We under stand that this McAb has important regulation effect on function of PMN, it shows that the specific combination with corresponding antigen on membrane is essential for function of PMN especially for mechanism of respiratory burst of activated PMN. PMID- 10453562 TI - [The association between vitamin D receptor gene polymorphisms, bone mineral density and osteocalcin in Chinese women]. AB - OBJECTIVE: To investigate the correlations between vitamin D receptor (VDR) gene polymorphisms, and bone mineral density (BMD) as well as osteocalcin. METHODS: We used PCR-RFLP and Southern hybridization analysis to determine VDR genotypes in 202 Chinese women. BMD was measured by dual-energy X-ray absorptiometry (DEXA) and serum osteocalcin level was determined by radioimmunoassay. Analysis of covariance was used to test differences in mean BMDs across genotypes. RESULTS: We realized that VDR genotype frequencies in Chinese women were apparently different from those of Caucasians. In group of postmenopausal women, "bb" or "aa" genotype was related to lower BMD in Femoral Neck and Trochanter. No association was found between VDR genotypes and the serum osteocalcin level. CONCLUSION: There was some association between VDR genotypes and BMD in Chinese women. Its function in predicting osteoporosis still requires further study. PMID- 10453563 TI - [Separation and purefied of calf osteocalcin, set up radioimmunoassay for osteocalcin]. AB - OBJECTIVE AND METHODS: We extracted and purified osteocalcin (BGP) from calf femur by gel filtration, FPLC and HPLC subsequently. The rabbits were immunized by this antigen and produced antibody. With this antibody (final 1: 35,000 dilution) we established RIA for BGP. This assay is stable and sensitive, the kd was 2.29 x 10(11). The intra assay variation was 2.1% and the inter assay variaton was 5.6%. It has no crossreaction with insulin, parathyroid hormone or calcitonin. RESULTS: The serum BGP values were detected in normal subjects and six kinds of bone metabolic diseases. In 81 normal subjects, the average serum BGP level was (5.3 +/- 1.5) ng/ml and in postmenopausal osteoporosis the serum BGP level [(6.2 +/- 1.9) ng/ml, n = 42] was higher than the normal (P < 0.05). The serum BGP levels were increased much in primary hyperparathyroidism [(12.8 +/ 7.0) ng/ml, n = 21, P < 0.001]. Increased values of BGP were also found in patients with chronic renal failure [(8.5 +/- 2.4) ng/ml, n = 18, P < 0.01] and Paget's disease [(6.7 +/- 2.2) ng/ml, n = 12, P < 0.05]. Decreased serum BGP values were found in Cushing syndrome [(2.9 +/- 0.9) ng/ml, n = 16, P < 0.001] and hypoparathyroidism [(3.4 +/- 1.1) ng/ml, n = 42, P < 0.001]. This result showed that in diseases with high bone turnover, the serum BGP levels were increased and in diseases with low bone turnover, the serum BGP levels were decreased. CONCLUSION: The data suggest that serum BGP is a sensitive marker for bone turnover. It is very improtant in studying bone metabolic diseases. PMID- 10453564 TI - [Further clinical analysis on the prognosis of chronic lymphocytic thyroiditis]. AB - OBJECTIVE AND METHODS: There have been many reports on the incidence of hypothyroidism and thyroid carcinoma in chronic lymphocytic thyroiditis patients, however, the incidence of thyroid carcinoma had been a widely debated issue. Therefore 300 cases of chronic lymphocytic thyroiditis from 1964 to 1995 were reviewed. There were 37 males and 263 females, and 52 had a pathological diagnosis of chronic lymphocytic thyroiditis. RESULTS: Among 300 patients, 2 had coexistent thyroid papillary carcinoma. The original symptom in 64.3% patients was hypothyroidism, and the total incidence of hypothyroidism was 76%. It was also indicated in this study that the occurrence of hypothyroidism in chronic lymphocytic thyroiditis patient was associated with the elevated level of antithyroglobulin antibody, enlargement and tenderness of thyroid gland. CONCLUSION: In our study, as the low incidence of coexisted thyroid carcinoma, with the high incidence of hypothyroidism it seemed to be that the elevated antithyroglobulin antibody could be a more effective factor in the development of hypothyroidism in chronic lymphocytic thyroiditis. PMID- 10453565 TI - [Preparation of McAb against lung cancer and its application in tumor radioimmunoimaging]. AB - OBJECTIVE: In order to tracing the lung cancer, A monoclonal antibody was produced. METHOD: The McAb, named SM-1, was specifically reacted with lung cancer confirmed by the immunopathological studies, then the McAb was digested into fragments by ficin and bromelain proteases. RESULTS: Both of the enzymatic methods were very effective and the immuno-reactivity of fragments was retained well. In our localization experiments, the T/NT ratio revealed that radioiodinated McAb and fragments could be specifically uptaked by the tumour tissue and the T/NT ratio was higher in fragments group than intact IgG. After injection of 131I-IgG and fragments, the xenografts of human lung cancer bearing in nude mice were visualized distinctly during 36-48 hrs. Eighteen patients with lung cancer received Tc-99m labeled McAb intravenously, 13 cases showed radioactive accumulation in the foci of tumours. CONCLUSION: These preliminary results suggested that SM-1 and its fragments might be a potential agents for radioimmunoimaging diagnosis of lung cancer. PMID- 10453566 TI - [MA891--an established spontaneous mouse mammary adenocarcinoma cell line and its immunologic characteristics]. AB - OBJECTIVE: To characterize the immunologic properties of an established mouse mammary tumor cell line MA891. METHODS: From a spontaneous mouse mammary adenocarcionma (TA2 MA891) of TA2 mouse origin, an in vitro passaged cell line MA891 was maintained in RPMI1640 with 10% new born calf serum. The immunogenecity of MA891 was studied by classical tumor transplantation rejection assay, generation of cytotoxic T lymphocytes (CTL) by syngeneic secondary mixed lymphocyte-tumor cell culture (MLTC) and cytotoxic assay of tumor infiltrating lymphocytes (TIL). RESULTS: MA891 maintained the high metastatic potential of the parental TA2 MA891 tumor passaged in vivo. Transplantation rejection studies indicated that amputation of a hind limb with growing tumor in the footpad did not protect the mice from a second subcutaneous challenge of the same tumor. The cytotoxic activities of both CTL and TIL were shown to be very weak. CONCLUSION: MA891 is a mouse tumor of very low immunogenecity. It can be used as a mouse tumor model for studying cancer metastasis in humans. PMID- 10453568 TI - [Detection of antineutrophil cytoplasmic antibodies for the values of Wegener's granulomatosis diagnosis and activity judgment]. AB - OBJECTIVE: To investigate the clinical significance of antineutrophil cytoplasmic antibodies (ANCA) in Wegener's granulomatosis (WG) and other diseases. METHOD: Indirect immunofluorescence was applied to detect serum ANCA of 204 patients and 48 normal controls. Enzyme-linked immunosorbent assay (ELISA) was used to detect antimyeloperoxidase (MPO) antibodies. RESULTS: The frequency of ANCA in sera of WG patients was 75% and significantly higher than those in sera of other groups (P < 0.05). ELISA demonstrated that MPO was the major antigen recognized by perinuclear ANCA (P-ANCA). Cytoplasmic ANCA (C-ANCA) was always positive and high titers in sera of active WG patients, but turned to negative or low titers in remission. CONCLUSION: These results suggested that C-ANCA was specific for WG and was a marker for disease activity in WG. P-ANCA wasn't specific for WG, but related to vasculitis. PMID- 10453567 TI - [Partial deletion of mitochondrial DNA in mitochondrial encephalomyopathies]. AB - OBJECTIVE: To characterize the deletions of mitochondrial DNA (mtDNA) in Chinese patients with Kearns-Sayre syndrome (KSS) and chronic progressive external ophthalmolegia (CPEO) and identify deletion mutations of mtDNA be the etiology of these diseases. METHODS: Patients and preparation of total DNA: Two patients with KSS and two with CPEO and ten with other mitochondrial myopathies or encephalomyopathies were determined by histological and biochemical assays. Total DNA was isolated from 100 mg to 150 mg of frozen muscle obtained by biopsy using the methods described by Zeviani et al. (1988). Preparation of mtDNA probes: MtDNA were isolated and purified according to Palva's method (1985). The mtDNA was linearized by digestion with the restriction endonuclease Pvu II and separated by electrophoresis through low melting agarose gel. The 16.5 kb DNA band was recovered from the gel and labeled with alpha-p32 dCTP by random primer labeling. Southern blot analysis: Portions of five micrograms total DNA obtained from the patients and controls were digested with Pvu II, EcoR I, Hind III And Xba I respectively. The digested DNA was separated by agarose gel electrophoresis and transferred to nitrocellulose membrane and then hybridized with P32 labeled mtDNA as previously described (Zhang, 1991). RESULTS: A large-scale deletions of mtDNA were identified in two patients with KSS and two patients with CEPO. The deletions ranged in size from 2.4 kb to 5.5 kb. The proportion of mutated mtDNA in each patients ranged from 54.6% to 84.6% of total mtDNA. No detectable deletions were found in mtDNA of ten patients with other mitochondrial myopathies or encepholomyopathies and normal controls. CONCLUSIONS: Deletions of mtDNA were found in all KSS and CEPO patients detected. These results supported The view point of the deletions are important causes of physical defect in KSS and CEPO. PMID- 10453569 TI - [The use of "self-locking" artificial vertebral prosthesis in surgical treatment for spinal tumor]. AB - OBJECTIVE: To evaluate the use of "self-locking" artificial vertebral prosthesis in surgical treatment for spinal tumor. METHODS: From 1992 to 1996, 14 cases with spinal tumor and paraplegia were treated with "self locking" artificial vertebral prosthesis designed by one of the authors (Ye Qibin) in our department. There were malignant giant cell tumor 2 cases; myeloma 4 cases; metastatic tumor 4 cases and one each with chondrosarcoma, malignant fibrous histocytoma, fibrosarcoma, hemangioma. After radical resection of the tumor and thorough decompression, the vertebral prosthesis was applied to reconstruct the spinal stability. Adjunctive chemotherapy or/and radiotherapy was given postoperatively. RESULTS: At the average follow-up period of 20.6 months 10 cases are still alive, 5 of them had survived for more than 2 years. 4 cases died with the average survival period of 19.8 months after operation. Almost all the patients obtained the improvement of the quality of the life with relative pain-free and intact neurologic function after operation. CONCLUSION: With "self-locking" artificial vertebral prosthesis after thorough anterior resection of spinal tumor, it is possible to provide solid internal fixation to reconstruct the spinal stability. PMID- 10453570 TI - [Application of rigid bronchoscopy in airway management after lobectomy of lung]. AB - OBJECTIVE: Application of rigid bronchoscopy in airway management after lobectomy of lung was recommended, and its significance was discussed. METHODS: Bronchoscopies were performed in three patients scheduled for lobectomy of lung undergoing epidural plus general anesthesia. Each patient was examined two times. The first examination was done after anesthetic induction and intubation. The second was given at the end of operation. RESULTS: Rigid bronchoscopy under general anesthesia could clearly show the inner structures of trachea, bronchi and location of carcinoma. Under direct version, the secretion and sludged blood in the trachea and bronchi after lobectomy could be scavenged. CONCLUSION: It was suggested that with the use of bronchoscopy the complications including postoperative atelectasis and bronchial-pleural fistulization could be reduced. PMID- 10453571 TI - [The usefulness of dye endoscopy in diagnosis esophageal diseases]. AB - OBJECTIVE: To investigate the usefulness of Dye endoscopy in the diagnosis of the esophageal disorders. METHODS: 87 cases with esophageal diseases underwent dye endoscopy examination with Toluidin-staining and Iodine-staining or double staining method. RESULTS: In the conventional group, the conformity rate of the endoscopy to biopsy results is 80%, but in the Dye one is 95% and the accurate rate of diagnostic is 100%. Under the guiding of Dye endoscopy, the occurrence rate of Ca cell on the incisal edge is 0%. CONCLUSION: The methods enable diffentinal diagnose of benign and malignant lesion, conforming the type of esophagitis. And it is more helpful to conform the area of surgical resection and when conventional endoscopy cannot satisfactorily diagnosis. We think it is a useful and simple diagnostic method for esophageal diseases. PMID- 10453574 TI - [Cytogenetic study for a child patient with mental retardation and eye multiple malformation]. AB - An cytogenetical study was carried out for a 7 years old patient with mental retardation, eye and other multiple malformation. An extra G-positive band was found in the region of 9q12. The possible relationship between this aberration and the abnormal figures was discussed. PMID- 10453572 TI - [Prophylactic effects of magnesium sulfate and ligustrazin on the hypoxic ischemic brain damage in neonatal rats]. AB - OBJECTIVE: A model in neonatal rats was established to study pathophysiology of perinatal hypoxic-ischemic brain damage. METHODS: 7-9 day old Wistar rats were subjected to unilateral carotied artery ligation followed by hypoxic state for 3 hours (10% O2 + 90% N2, at 37 degrees C). Magnesium sulfate (0.5 mg/gBW) and ligustrazin (0.1 mg/gBW) was separately given a 30 min before hypoxia. Superoxide dismutase (SOD) and malonic dialdehyde (MDA) in the cerebral cortex and serum MDA were measured immediately after hypoxia. Neuropathologic examination was made in 4 weeks after hypoxia. RESULTS: It was found that SOD and MDA in the cerebral cortex and serum MDA in the hypoxicischemic group were significantly increased in comparision with the normal control group (P < 0.01), and that all these parameters either in the magnesium sulfate group or in the ligustrazin group were lower than in the hypoxic-ischemic group (P < 0.05). The hypoxic-ischemic group showed that there were neuronal degenerations in the gray matter, hippocampus and cerebellum which were reduced in the two treatment groups. CONCLUSION: Our results indicate that oxide free radical formation is one of the pathogenic factors of perinatal hypoxic-ischemic brain damage and magnesium sulfate and ligustrazin reduce hypoxicischemic brain damage due to indirect anti-oxidation. PMID- 10453573 TI - [Observation on the content of plasma lipoperoxide and serum superoxide dismutase in NIDDM nephrosis patients]. AB - OBJECTIVE: In order to explore the changes of MDA, SOD in each period of diabetic nephrosis patients, the contents of MDA and SOD from 40 cases of NIDDM nephrosis patients in various stages and 15 cases of normal persons (control group) were measured. METHODS: By using method of sulpho-barbiamino acid to determine the concentration of serum MDA (the reagents are provided by Juli Biomedical Engineering Institute of Nanjing). By using method of radioimmunization to determine the concentration of SOD (The reagents are provided by Huaqing Biological Technic Institute of Beijing), blood sugar, T-cho, TG, Bun, Ccr, UAE, etc. in serum were simutaneouslly determined. By adopting statistical method to analyse the mean difference in each groups and the interrelation of each data. RESULTS: The results show that the content of MDA in each groups exceeds normal control group and the content of MDA of the group of diabetic kidney failure (DKF) exceeds group of diabetic kidney early stage (DKE) obviously (P < 0.01). Only trace of MDA and SOD can be detected in the liquid of peritoneal dialysis in group of DKF. The content of SOD in each groups exceeds normal control group without exception and the content of SOD in group of DKF exceeds that of the group of diabetic kidney function injury stage obviously (P < 0.01). CONCLUSION: Our results suggest that content of SOD might have some clinical value for the diagnosis and prognosis in NIDDM nephrosis and situation of kidney function injury. PMID- 10453575 TI - [The curative effects of anti-TNF monoclonal antibody in E. coli infected mice]. AB - OBJECTIVE: To evaluate effects of monoclonal antibody (McAb) against TNFa on E. coli infected mice. METHODS: 30 mice (10 week old, Kunming) were divided equally into three groups. The first group, as a control, 200 microliters saline (NS)/mouse was injected intravenously; Second group (untreated group), only E. coli (10(7) organisms/200 microliters NS) were injected intra-abdominaly; Third group (treated group), E. coli (10(7) organisms) were injected intra-abdominal and McAb against TNF alpha 2 mg/kg dissolved in 200 microliters NS were injected intravenously. After 24 h, observed the survival rate, compared the serum TNF level in blood and investigated pathology of intestine, lung and liver. RESULTS: There was a higher survival rate in treated group, with the serum TNF level lowered significantly, and the untreated mice had severe pathologic changes in viscera. CONCLUSION: Using anti-TNF alpha McAb was effective in reducing mortality rate in mice after infected with E. coli, but could not prevent the pathologic changes. PMID- 10453576 TI - Treatment of complications due to peritoneal dialysis for chronic renal failure with traditional Chinese medicine. AB - In this paper, the experience in the treatment of complications due to continuous ambulatory peritoneal dialysis for chronic renal failure with traditional Chinese medicine (TCM) is reported. Modified Renshen Yangrong Tang (Ginseng Nutrition Decoction) was used for anorexia and hypoproteinemia; modified Xiangsha Liujunzi Tang (Decoction of Cyperus and Amomum with Six Noble Ingredients) for abdominal pain and distension; modified Da Chaihu Tang (Major Bupleurum Decoction) for peritonitis; modified Shenling Baizhu San (Powder of Ginseng, Poria and Atractylodes) for diarrhea due to insufficiency of the spleen with abundance of dampness; Lizhong Tang (Decoction for Regulating the Function of Middle-jiao) and modified Sishen Wan (Pills of Four Miraculous Drugs) for insufficiency of both the spleen and the kidney; Siwu Tang (Decoction of Four Ingredients) added with other drugs for cutaneous pruritus, and Guishao Sijunzi Tang (Decoction of Four Noble Drugs added with Chinese Angelica Root and white Peony Root) for renal anemia. The therapeutic principles of invigorating the liver and kidney, strengthening the bones and muscles, and promoting blood circulation to eliminate blood stasis were adopted in the treatment of renal osteopathy, and the therapeutic principles of invigorating the liver and kidney, expelling phlegm and resolving dampness, and promoting blood circulation to eliminate blood stasis in the treatment of hyperlipemia. Shen Tekang capsules (capsules for improving the renal function) was administered to patients for strengthening the viability and improving the nutrition state, and the recipe for treating renal function failure (both formulated by the authors) for improving the renal function so as to decrease the frequency and duration of dialysis. PMID- 10453577 TI - TCM differential treatment of 57 cases of chronic gastritis complicated by ulcerative colitis. AB - 57 cases of chronic gastritis complicated by ulcerative colitis were clinically observed. According to the TCM theory of differentiation of symptoms and signs, 4 types of the disease were divided and treated: 1) stagnation of qi in the liver, 2) dampness and heat in gastrointestinal tract, 3) asthenic cold in the spleen and stomach, and 4) coexistence of cold and heat. The therapeutic effects of chronic gastritis and ulcerative colitis were evaluated: 23 cases (40.3%) of chronic gastritis were cured and 28 cases (49.1%) improved; 25 cases (43.8%) of ulcerative colitis were cured and 28 cases (49.1%) improved. The effective rates were 89.5% and 93.0% respectively. The correlation between the therapeutic effects for these two diseases further demonstrates that their pathological mechanisms are closely interrelated, indicating that it is important to have these two diseases treated simultaneously. PMID- 10453578 TI - A clinical study on acupuncture for prevention and treatment of toxic side effects during radiotherapy and chemotherapy. PMID- 10453579 TI - Therapeutic effect of needle warming through moxibustion at twelve shu points on rheumatoid arthritis. PMID- 10453580 TI - Clinical observations on acupuncture treatment of pseudobulbar palsy--a report of 325 cases. PMID- 10453581 TI - A clinical trial on yu cong tang in treatment of senile dementia. AB - Yu Cong Tang ([symbol: see text]), a TCM decoction, was used to treat senile dementia. It has the action of invigorating the spleen, reinforcing the kidney, nourishing yin, strengthening yang, improving blood circulation, dredging the channels, removing phlegm and restoring consciousness. CT, EEG, REG, Hb, PaO2, Tch, TG and 17-OHCS were tested, showing significant differences (P < 0.05 or < 0.01) before and after the treatment. Remarkable improvement was found in clinical symptoms, and the effective rate was 94.1%. Significant differences were found between the TCM and western medicine (WM) groups in symptom improvement (P < 0.05-0.01) and therapeutic results (P < 0.001). PMID- 10453582 TI - Effects of acupuncture on the levels of endothelin, TXB2, and 6-keto-PGF1 alpha in apoplexy patients. AB - In order to delve into the mechanism governing the treatment of apoplexy by acupuncture at yangming channel points as main points, we observed the changes in the endothelin (ET) level in plasma, TXB2 and 6-Keto-PGF1 alpha levels in urine in convalescent apoplexy patients during acupuncture treatment. The results showed that the ET level in plasma in convalescent apoplexy patients was significantly higher than that in healthy subjects (P < 0.05), and the ET level in plasma in patients was decreased after one course of acupuncture treatment. It was found that before treatment the TXB2 level in urine in apoplexy patients was significantly higher than in healthy subjects, and the 6-Keto-PGF1 alpha level in urine in the patients was significantly lower than that in healthy subjects, with an increased ratio of TXB2 to 6-Keto-PGF1 alpha. After acupuncture treatment, the TXB2 level in urine was lowered with a decrease in the ratio of TXB2 to 6-Keto PGF1 alpha. All this indicated that one of the mechanisms governing acupuncture treatment of apoplexy acupuncture at yangming channel points as main points was that acupuncture could produce therapeutic effects by adjusting the imbalance of important vaso-active substances, ET, TXA2, and PGI2. PMID- 10453583 TI - 80 cases of peripheral facial paralysis treated by acupuncture with vibrating shallow insertion. AB - Eighty cases of peripheral facial paralysis were treated by acupuncture with vibrating shallow insertion. The total effective rate was 98.75% and cure rate 88.75%. There was no significant difference in therapeutic effect as compared with the conventional electroacupuncture method (P > 0.5), suggesting that shallow puncture by vibratings can also yield satisfactory therapeutic results. PMID- 10453584 TI - Clinical experience in acupuncture treatment of obesity. AB - Obesity manifests itself as derangement of qi and blood with disorder of the defensive qi in particular to be a causative factor of primary importance, meanwhile, deficiency of the primary qi in Sanjiao may also contribute to it. Fundamentally, its pathological changes are also related to dysfunction of the spleen and stomach. According to syndrome differentiation, obesity can be classified into three types, namely, heat in the stomach and intestine, spleen and stomach qi deficiency, and deficiency of primary qi. The main methods of treatment include clearing and reducing heat in the stomach and intestine; regulating qi in the fu organs and promoting water metabolism; balancing and regulating qi and blood, yin and yang, and removing stagnation of defensive qi. PMID- 10453585 TI - Treatment of spinal-cord-type cervical spondylosis by Chinese massotherapy. PMID- 10453586 TI - Effects of electro-acupuncture on somatostatin and pancreatic polypeptide in ischemic cerebrovascular diseases. AB - The levels of somatostatin (SS) in CSF and blood and pancreatic polypeptide (PP) in plasma were measured by radioimmunoassay in 64 patients with acute ischemic cerebrovascular diseases (ICVD), randomly divided into two groups: group 1 (n = 31, both electro-acupuncture and routine treatments given) and group 2 (n = 33, routine treatment) and 26 non-ICVD patients were used as controls. The points of electro-acupuncture were Quchi (LI 12), Waiguan (SJ 5) and Huantiao (GB 30) and Zusanli (St 36). After a course of treatment, the SS levels in plasma and CSF were significantly increased in the patients of group 1 with good result and their plasma PP level had no significant change. In the patients with poor result, however, the PP level was significantly decreased. The results suggested that electro-acupuncture might play an active role in alleviating the SS metabolic disturbance in CNS of ICVD patients. PMID- 10453588 TI - A review on traditional Chinese medicine in prevention and treatment of multiple sclerosis. PMID- 10453587 TI - An experimental study of electro-acupuncture on auditory impairment caused by kanamycin in guinea pigs. AB - Frequency following response (FFR) and auditory brain stem evoked potential response (ABR) were used to determine the auditory acuity in evaluating the effect of electro-acupuncture treatment of kanamycin-induced auditory impairment in guinea pigs. The succinate dehydrogenase (SDH) activity and morphological changes of the inner ear receptors were examined under the light and scanning electron microscope in cochlear spread preparations. The results showed that 1) electro-acupuncture was effective but no significant differences were found among the stimulating wave forms; 2) Tinggong (SI 19), Yifeng (SJ 17), Shenshu (UB 23), Sanyinjiao (Sp 6), Zhubin (K 9) and Waiguan (SJ 5) are all effective acupoints, especially the combination of Tinggong (SI 19), Sanyinjiao (Sp 6) and Zhubin (K 9) acupoints; 3) improvement in the cochlear function and excitability of the cortical and lower auditory center and increase of the mitochondrial SDH activity and energy supply in hair cells might contribute to the mechanism of the treatment. PMID- 10453589 TI - Ma xing er san tang for treatment of cor pulmonale at acute onset stage--a report of 250 cases. PMID- 10453590 TI - A clinical trial of tang shen ning for treatment of diabetic nephropathy. AB - This paper reports the clinical trial of Tang Shen Ning ([symbol: see text], TSN) for treating diabetic nephropathy (incipient and clinical, as divided by Mogensen). The results showed that the total effective rate in treatment group (TSN + western medicine) was 90.0%, and that in the control group (simply with western medicine), 56.7%. TSN plays important roles in decreasing proteinuria and improving renal functions. PMID- 10453591 TI - Acupuncture treatment of hysteric aphonia--a report of 27 cases. PMID- 10453592 TI - 60 cases of shoulder-arm syndrome treated by electroacupuncture at Bingfeng (SI 12). PMID- 10453593 TI - 300 cases of menopausal syndrome treated by acupuncture. PMID- 10453594 TI - 32 cases of femoral adductors syndrome treated by electroacupuncture and moxibustion. PMID- 10453595 TI - Application of acupuncture and moxibustion for keeping shape. AB - On the basis of TCM diferentiation, the authors have treated 359 adult female cases of non-obesity with undesirable body shape by combined application of body acupuncture and moxibustion and auricular acupuncture, and achieved quite good remoulding and orthopedic results, suggesting that acupuncture and moxibustion can very effectively regulate the somatotypic indexes of body weight, circumference of the chest, loin, hip and thigh, the ratio of the loin to hip, sebum thickness, obesity degree, body mass index and body fat percentage of the cases in the overweight group and orthopedic groups 1, 2. It is therefore concluded that acupuncture and moxibustion is a good therapy for obtaining a graceful body shape. PMID- 10453596 TI - Selection of acupoints by Doppler sound spectrogram for treatment of insufficient blood supply of cerebral basilar artery. PMID- 10453597 TI - Experience in the point-selection for electro-acupuncture. PMID- 10453598 TI - 48 cases of anxiety syndrome treated by massage. PMID- 10453599 TI - A clinical investigation on massage for prevention and treatment of recurrent respiratory tract infection in children. AB - The TCM therapeutic principle of "eliminating the pathogenic factors by supporting the healthy energy" was adopted in massage to treat and prevent recurrent respiratory tract infection of children. Susceptible and healthy children of the same age were used as controls. As a result, the therapeutic effect of the treatment group is significantly better than that of the controls, with all of the immunologic indexes being approximately normal when the patients were reexamined 3 and 6 months after the massage. Statistically, the difference was significant (P < 0.01). Massage has been proved very helpful in improving the general constitution, enhancing the immune functions, preventing and treating the condition, and in health care as well. PMID- 10453600 TI - The effects of radix Salviae miltiorrhizae on lipid accumulation of peroxidized low density lipoprotein in mouse peritoneal macrophages--lipid analysis and morphological studies. AB - Mouse peritoneal macrophages were incubated in DMEM with pox-LDL and Rradix Salviae Miltiorrhizae (RSM) to investigate the effects of RSM on the internalization of peroxidized low density lipoprotein (pox-LDL) by using lipid analysis and electron microscopy. Lipid peroxide (LPO) concentrations were increased slightly in the medium after incubation of macrophages with normal LDL (n-LDL), while decreased significantly in the media after incubation of macrophages with pox-LDL. In the three groups with pox-LDL, it could be found that there was a dose-dependent decrease of concentrations of LPO and total cholesterol (TCH) in the two RSM groups, and the decrease in the two RSM groups was much greater than in the group without RSM. RSM accelerated a more decrease of LPO than cholesterol contents in the media containing pox-LDL. The ultrastructural studies also showed that RSM induced the accumulation of lipid droplets in the cytoplasm of mouse peritoneal macrophages. The results suggested that RSM could accelerate the phagocytosis and degradation of pox-LDL by macrophages. PMID- 10453601 TI - The effect of bu shen sheng xue fang on beta-thalassemia at gene level. AB - Isoelectrofocusing electrophoresis, PCR-SSCP analysis with DNA direct sequencing and RT-PCR were used to analyze the constituents of hemoglobin, determine the pattern of globin gene mutation, and detect its mRNA transcription extent. The results indicated that Bu Shen Sheng Xue Fang (BSSXF) significantly increased the contents of hemoglobin (Hb) and fetal hemoglobin (HbF), and the chain ratio in patients with beta-thalassemia. The drug is markedly effective in heterozygous, but not effective in homozygous or genetic compound patients. It was suggested that the beta-globin gene defects were compensated by elevating the gamma/beta + gamma ratio, promoting the transcription and expression of gamma-globin gene in synthesizing HbF. PMID- 10453602 TI - Treatment of tennis elbow by acupuncture and moxibustion. PMID- 10453603 TI - Literature research on point injection with Chinese Angelica liquor. PMID- 10453604 TI - Dr. Zhou Zhongying's experience in TCM treatment of Parkinson's disease. PMID- 10453605 TI - A comparative observation on treatment of heart failure due to diastolic insufficiency of the left ventricle--a report of 43 cases. AB - 43 cases of heart failure due to diastolic insufficiency of the left ventricle were treated with TCM differential treatment as the treatment group, the total effective rate being 86.0%. Statistically significant difference (P < 0.05) in the therapeutic effects was found between the treatment group and control (25 cases) group. Improvement of the left ventricular diastole in the treatment group was more obvious than that of the control group (P < 0.01), while contractility of the left ventricle was also improved to a certain extent. PMID- 10453606 TI - TCM differential treatment of child hood persisting pneumonia--a report of 52 cases. AB - According to TCM differentiation of the symptoms and signs, 52 children with presisting pneumonia were divided into the following four types: 1) deficiency of body fluid due to accumulation of heat in the lung, 2) earth failing to generate metal, 3) deficiency of the Ying and Wei systems, and 4) excessiveness of pathogens due to qi deficiency. After treatment for 2-3 weeks, 40-cases were cured, 8 improved, and 4 ineffective. PMID- 10453607 TI - Treatment of atrophic cholecystitis by regulating the function of the spleen--a report of 50 cases. AB - Fifty cases of atrophic cholecystitis were treated mainly by regulation of the function of the spleen. Of them, 21 cases were cured, 18 markedly effective, and 7 effective. The total effective rate was 92.0%. By comparison of results of ultrasonography B performed before and after treatment, it was shown that both the longitudinal and transverse inner diameters of gallbladder cross section increased evidently, and the condition of atrophy was improved remarkably after treatment. PMID- 10453608 TI - Clinical observation on acupuncture treatment of uroschesis. PMID- 10453609 TI - Observation on frequency spectrum of electrogastrogram (EGG) in acupuncture treatment of functional dyspepsia. PMID- 10453610 TI - 23 cases of chronic nonspecific ulcerative colitis treated by acupuncture and moxibustion. PMID- 10453612 TI - 106 cases of non-suppurative costal chondritis treated by acupuncture at xuanzhong point. PMID- 10453611 TI - Clinical observation on the therapeutic effects of wrist-ankle acupuncture in treatment of pain of various origins. PMID- 10453613 TI - 32 cases of tennis elbow treated by the three-needle therapy on the elbow. PMID- 10453614 TI - Acupuncture treatment of phantom limb pain--a report of 9 cases. PMID- 10453615 TI - 14 cases of child bronchial asthma treated by auricular plaster and meridian instrument. PMID- 10453616 TI - 228 cases of painful swelling of the yang-heat type treated by compress with xiao zhong san. PMID- 10453617 TI - A clinical study on bing gan ling oral liquid for treatment of hepatitis C. AB - Based on the principles of cooling the blood to expel the pathogenic toxins and regulating the function of liver and activating the spleen in traditional Chinese medicine, 60 cases of chronic hepatitis C were treated with Bing Gan Ling (BGL), an oral liquid, which is composed mainly of Shuiniujiao (Cornu Bubali), Huzhang (Rhizoma Polygoni Cuspidati), Chishao (Radix Paeoniae Rubra), and Huangqi (Radix Astragali), etc. The total effective rate was 86.7%, which is considerably better than that of the control group (P < 0.01). The animal experiments and pathological studies suggest that the above-mentioned prescription has marked action of protecting the liver and lowering the activity of enzymes; and that it can ameliorate the acute liver injury induced by D-galactosamine. PMID- 10453618 TI - Clinical observation on treatment of 83 cases of posthemiplegic omalgia. AB - An analysis on 83 cases of posthemiplegic omalgia (shoulder pain) shows that the pathogenesis of the pain is closely related to the improper passive movement at the early stage of hemiplegia (62.7%). The large range of passive movement is a dangerous factor leading to omalgia. In the study of upper extremity complications, the incidence of shoulder-hand syndrome is relatively high (42.2%), and it is often accompanied by hand swelling (83.1%). The authors suggest that painless movement of the shoulder joint should be limited in a range of 90-120 degrees, massage be carried out immediately after acupuncture, and the affected upper extremity be moved passively during the needle retention. This therapeutic method is definitely effective for pasthemiplegic omalgia. PMID- 10453619 TI - Treatment of 40 cases of hydroceles with massage at qichong (St 30). PMID- 10453620 TI - Effects of electroacupuncture and twirling reinforcing-reducing manipulations on volume of microcirculatory blood flow in cerebral pia mater. AB - Effects of electroacupuncture (EA) and routine acupuncture with twirling reinforcing and reducing manipulation of the needle (RA) both at Zusanli point (St 36) on volume of microcirculatory blood flow in the cerebral pia mater were observed by fenestration of the cranial bone and laser Doppler microcirculatory blood flow analyser. Results showed that both RA and EA could increase the volume of microcirculatory blood flow in the cerebral pia mater; and that the increase in the EA group was superior to that in RA group. This suggests that a moderate and effective stimulation is a key to the production of a regulative effect on the organism. PMID- 10453621 TI - On the indications of acupuncture and moxibustion. PMID- 10453622 TI - Advances in TCM treatment of hyperprolactinemia. AB - In recent years, rapid progress has been seen in the treatment of hyperprolactinemia with TCM and materia medica. The old pattern of simply using bromocryptine to treat the disease has been changed. Most of the cases reported in this paper were primary hyperprolactinemia, with other primary diseases excluded. As TCM treatment produces definite therapeutic effects with few side effects, the methods above reported are worth recommending. PMID- 10453623 TI - Respiratory protection on offshore drilling rigs. PMID- 10453624 TI - PAT Program. Background and current status. PMID- 10453625 TI - Occupational heat exposure. PMID- 10453626 TI - Controlling silica dust from foundry casting-cleaning operations. National Institute for Occupational Safety and Health. PMID- 10453628 TI - Portable gas chromatographs. PMID- 10453630 TI - Occupational exposure to styrene in Norway, 1972-1996. AB - Occupational exposure to styrene has been of special concern in Norway since the beginning of the 1970s, especially the exposure in the reinforced plastic industry. Campaigns have been carried out by the Norwegian Directorate of Labour Inspection and various measures have been set forth to reduce the workers' exposure to styrene. Alongside the campaigns, extensive monitoring programs were carried out. The measurement results are compiled in the database EXPO at the National Institute of Occupational Health (Norway). This article presents a statistical analysis of the measurement data to assess the occupational exposure to styrene in Norway since the 1970s. Results from 7,011 measurements carried out in 234 enterprises covering the period 1972-1996 are included in the database. The analyses show occupational exposure to styrene of 33.0 ppm (median) (mean: 49.4 ppm) for this period. The analyses show a decrease from a median of 62 ppm in the 1970s to 7.1 ppm in the 1990s. The analyses do not disclose any crucial impact from any single preventive measure implemented by the authorities to reduce exposure level. The reduction in the level is probably due to a contribution from different measures. The decline in the styrene exposure coincides with a similar reduction in other countries, and it may be that international concern and technical improvements in the manufacturing process have been an important factor in the exposure level reduction in the Norwegian plastic reinforced industry as well. PMID- 10453631 TI - Conjugation of 7-oxodehydroabietic acid to lysine, a haptenation mechanism for an oxidized resin acid with dermal sensitizing properties. AB - This article explores protein conjugation of 7-oxodehydroabietic acid, a resin acid found in both aerosol from soldering with rosin flux and in rosin solids. In a murine model, conjugation (haptenation) of resin acids to proteins is required to generate antibodies against rosin. Hydroperoxy resin acids are dermal sensitizers, with haptenation thought to occur via radical mechanisms. Dermal sensitization to 7-oxodehydroabietic acid has been observed, although no radical haptenation mechanism has been proposed to explain the sensitizing properties of this compound. Conjugation of L-lysine to 7-oxodehydroabietic acid was predicted, with a Schiff base (or imine) linkage formed between C-7 of the resin acid and a free amino group of lysine. Fast atom bombardment mass spectrometry provided evidence of the conjugate; a small peak was seen for the conjugate (M+H)+ ion in aqueous ethanol with 20 mM concentrations of the free resin and amino acids. A larger conjugate peak was observed with addition of tertiary amine as a mild basic catalyst, and the intensity of the conjugate peak exceeded that of the precursors upon replacement of the ethanol with benzene. Resin acids accumulate in the plasma membrane, a non-aqueous environment apparently conducive to conjugation of 7-oxodehydroabietic acid with lysine side chains of membrane proteins. The result would be dehydroabietic acid covalently bound to protein, which could lead to interaction with immune cells having resin acid specificity. The haptenation mechanism presented may be involved in allergic contact dermatitis and occupational asthma observed from exposure to resin acid solids and aerosols. As sampling and analytical methods have been previously demonstrated for 7-oxodehydroabietic acid, this compound may be a useful exposure marker with relevance to negative health effects such as occupational asthma. PMID- 10453632 TI - An evaluation of worker lead exposures and cleaning effectiveness during removal of deteriorated lead-based paint. AB - We evaluated worker lead exposures and cleaning effectiveness during initial cleanup of 19th-century buildings with highly deteriorated lead-based paint. Eighteen rooms of similar size and condition in two university-owned buildings were selected for a pilot project to compare three methods for removing loose paint, paint chips, and dust. The methods used were: dry scraping followed by dry sweeping (no engineering or work practice controls); wet scraping and high efficiency particulate air (HEPA) vacuuming; and the latter method with the addition of a portable HEPA-filtered exhaust fan in the room providing about 40 air changes per hour. The final step for all methods was wet-mopping once with tri-sodium phosphate solution. During a single day 18 rooms were cleaned; each of three two-person work crews cleaned six rooms, two with each method. Air and surface samples were collected before, during, and after cleaning. All of the methods were potentially hazardous to workers: 44 percent of the method-based exposures (range: 5.0-360 micrograms/m3) and one of five full-shift exposures exceeded the OSHA PEL (range 9.4-110 micrograms/m3). Lowest worker exposures were during the wet scraping and vacuuming method (mean: 24 micrograms/m3). Providing general ventilation in rooms did not reduce worker exposures and appeared to increase them (mean: 73 micrograms/m3). Overall, the mean floor surface lead levels were reduced 50 percent after cleaning (from 2,600 to 1,300 micrograms/ft2), but the effectiveness of the three methods in reducing floor lead levels did not differ significantly. Overall, the method, mean paint lead concentration, pre-cleaning surface lead concentration, and work crew were significantly associated with the mean worker exposures during cleaning (p = 0.023), but not with the post-cleaning surface lead concentrations (p = 0.13). PMID- 10453633 TI - Measurements of mineral oil mist, hydrocarbon vapor, and noise in engine rooms of ships. AB - The purpose of this study was to determine the concentrations of oil mist and hydrocarbon vapor to which marine engineers are currently exposed. Measurements were also taken of the sound level in the engine room and the control rooms. Area mist concentration measurements were performed in 21 ferries, 2 cargo ships, and 1 westamaran (an express ship with two keels). Measurements were also performed for four different tasks where exposures above area level were expected. The area level of oil mist in the engine rooms of the different ships varied from not detectable to 0.53 mg/m3 (mean 0.24 mg/m3). The levels of hydrocarbons in the different ships varied from 0.2 to 14.5 mg/m3. The sound level varied from 96 to 108 dB(A) in the engine rooms, and from 70 to 90 dB(A) in the control room. When compared to other studies, it is supposed that the exposure to noise and mineral oil mist in the engine rooms of ships may represent a risk of adverse health effects for workers on Norwegian ships. The sound level may cause neurogenic hearing loss when appropriate hearing protection is not used. PMID- 10453634 TI - Chemical components of shredded paper insulation: a preliminary study. AB - We conducted an evaluation of shredded paper insulation to identify potentially toxic components. The study was to provide a preliminary characterization of a few samples of insulation currently in use. The following samples were analyzed: previously produced insulation (PPI) containing fire retardants, shredded recycled paper (PPI feedstock), freshly produced insulation (FPI), and insulation which had been installed in a residence (II). Volatile constituents were analyzed by GC-MS from headspace air of samples held at room temperature or heated to 90 degrees C. Extractable constituents were sampled by extracting with methylene chloride, and analyzing by GC-MS. Formaldehyde analysis was done according to EPA Method TO11. Headspace air at room temperature contained no detectable quantities of volatile constituents for any sample measured. In headspace air at 90 degrees C, only PPI contained traces of aliphatic and aromatic hydrocarbons and higher aldehydes, and FPI traces of toluene. Extracts of PPI contained traces of octadecadienoic acid methyl ester and aliphatic and aromatic hydrocarbons and higher aldehydes. Extracts of PPI feedstock contained traces of a substituted cyclohexenecarboxylic acid. FPI contained extractable diethyl phthalate (30-50 micrograms/g). Extracts of II contained traces of methyl palmitate, an octadecenoic acid methyl ester, and a phthalate plasticizer. No formaldehyde was detected. PPI was composed of approximately 98 percent paper fiber and 2 percent pre-gelatinized starch. PPI samples agglomerated together with less than 0.01 percent separating from clumps as fine dust. Boron and sodium were expected and confirmed because they were added to PPI and FPI as fire retardants. Chromium, copper, iron, potassium, magnesium, manganese, phosphorus, and silicon were present at detectable concentrations. Study calculations indicate that an occupant would have to completely consume all the fine particles produced from 3.3 kg of insulation per day to have an intake of boron equivalent to the EPA RfD. No other constituent appeared to be present even close to toxicologically relevant amounts. PMID- 10453635 TI - Peptide libraries in cellular immune recognition. PMID- 10453636 TI - Antigen sequence- and library-based mapping of linear and discontinuous protein protein-interaction sites by spot synthesis. AB - The knowledge (antigen-derived peptide scans)- and library (de novo)-based mapping of linear and discontinuous antibody epitopes as well as protein-protein contact sites in general by spot synthesis now is a well established technique. Due to its automation, this technique also promises great potential for applications in functional genomics. It should help to elucidate the complex network of interacting protein molecules involved in signal transduction events (Adam-klages et al. 1996; Hoffmuller et al. 1999). Although little chemistry is involved in the preparation of peptide scans or libraries and the synthesis procedure is relatively simple, the laboratories of immunologists or molecular biologists are often not equipped to perform spot synthesis. In this case scans or libraries can be purchased from commercial suppliers. PMID- 10453637 TI - The two hybrid toolbox. PMID- 10453638 TI - Evolutionary approaches to protein engineering. PMID- 10453639 TI - Phage display of combinatorial peptide and protein libraries and their applications in biology and chemistry. PMID- 10453640 TI - In vitro selection methods for screening of peptide and protein libraries. PMID- 10453641 TI - Aptamers as tools in molecular biology and immunology. AB - We have listed and described recent promising developments in the field of aptamer research. The properties and the application potential of aptamers propose an exciting future for aptamers either in the clinic or as research tools for various purposes. We have reviewed exciting examples in which the SELEX technology was applied to obtain promising tools that may help to facilitate our understanding of biological processes and to interfere at distinct points in signal transduction cascades. High affinities and specificities of aptamer/target interactions can now routinely be achieved. Furthermore, a wide spectrum of chemical modifications of nucleotides is known which greatly increase the stability of RNA molecules in biological materials, considerably enhancing their application potential. The aptamer technology shows that the combination of organic synthesis and molecular biology can contribute to interesting and promising new drug leads, which may very soon find their way into daily clinical practice or onto the laboratory benches of many researchers in the life sciences. PMID- 10453642 TI - In vitro selection of nucleic acid enzymes. PMID- 10453643 TI - Dynamic combinatorial chemistry: evolutionary formation and screening of molecular libraries. PMID- 10453644 TI - Evolutionary biotechnology--reflections and perspectives. PMID- 10453645 TI - Biology of immunoreceptor tyrosine-based inhibition motif-bearing molecules. PMID- 10453646 TI - Negative regulation of hematopoietic cell activation and proliferation by Fc gamma RIIB. PMID- 10453647 TI - The role of the SRC homology 2-containing inositol 5'-phosphatase in Fc epsilon R1-induced signaling. PMID- 10453648 TI - The unexpected complexity of Fc gamma RIIB signal transduction. PMID- 10453650 TI - HLA-specific and non-HLA-specific human NK receptors. PMID- 10453649 TI - Regulation of B cell antigen receptor signaling by the Lyn/CD22/SHP1 pathway. PMID- 10453651 TI - Regulation of NK cell functions through interaction of the CD94/NKG2 receptors with the nonclassical class I molecule HLA-E. PMID- 10453652 TI - Inhibitory role of murine Ly49 lectin-like receptors on natural killer cells. PMID- 10453653 TI - gp49: an Ig-like receptor with inhibitory properties on mast cells and natural killer cells. PMID- 10453654 TI - A novel family of inhibitory receptors for HLA class I molecules that modulate function of lymphoid and myeloid cells. PMID- 10453655 TI - Interactions of LIRs, a family of immunoreceptors expressed in myeloid and lymphoid cells, with viral and cellular MHC class I antigens. PMID- 10453656 TI - Paired immunoglobulin-like receptors of activating and inhibitory types. PMID- 10453657 TI - LAIR-1, a widely distributed human ITIM-bearing receptor on hematopoietic cells. PMID- 10453658 TI - The mast cell function-associated antigen, a new member of the ITIM family. PMID- 10453659 TI - The enigma of activating isoforms of ITIM-bearing molecules. PMID- 10453660 TI - Educational forum a laudable undertaking. PMID- 10453661 TI - Differential diagnosis and treatment strategies for biologic complications and failing oral implants: a review of the literature. AB - The aim of this article was to review the literature on differential diagnosis and treatment of biologic complications and failing implants. All types of publications, with the exception of abstracts, published in English up to December 1998, were included. A multi-layered search strategy was used. Controlled clinical trials (CCTs) were searched in the Cochrane Oral Health Group's Specialized Register of Trials. This database contains all CCTs identified in MEDLINE and EMBASE. PubMed was searched using various key words and the "related articles" feature. All identified publications were obtained and none were excluded. Infection, impaired healing, and overload are considered the major etiologic factors for the loss of oral implants. Only a few clinical and animal investigations were found that tested the validity of the proposed therapeutic approaches. The treatment of failing implants is still based mainly on empirical considerations, often derived from periodontal research, from data extrapolated from in vitro findings, or from anecdotal case reports performed on a trial-and-error basis. PMID- 10453662 TI - Comparison of interface relationships between implant components for laser-welded titanium frameworks and standard cast frameworks. AB - With the introduction of new techniques for the fabrication of frameworks for implant-supported prostheses comes the need to understand how the components used compare to those used for conventional cast frameworks. The relationship of measured machining tolerances between conventional implant components and those components used for stereo laser-welded implant frameworks was determined using a standardized protocol. Statistically significant differences in the horizontal interface relationship were found between paired implant components, which had a mean range from 23.1 to 51.7 microns. From a laboratory and clinical perspective, machining tolerances of implant components represent a variable associated with their manufacturing, which can ultimately affect the fit of a completed prosthesis. PMID- 10453663 TI - Clinical application of narrow Branemark System implants for single-tooth restorations. AB - Replacing small, single incisors with implants can be esthetically challenging and difficult because of the limited amount of bone. In this investigation, 3.0 mm-diameter implants were used to support 30 single maxillary and mandibular incisors in 21 patients. The implants have been in function for 3 to 7 years, and 29 are still stable. Only 2 complications in the mandibular incisor region have occurred; 1 implant fractured (after 5 years of function) and 1 prosthesis was replaced. The overall success rate is 96.7%. The favorable results and esthetic appearance achieved suggest that replacing small incisors where light occlusal forces are present with narrower implants is a feasible treatment option. PMID- 10453664 TI - Occlusal contacts of edentulous patients with mandibular hybrid dentures opposing maxillary complete dentures. AB - The purpose of this study was to evaluate differences between the occlusal contacts of edentulous patients with mandibular implant-supported hybrid dentures opposing maxillary complete dentures and those of patients with conventional maxillary and mandibular complete dentures. Six parameters related to occlusion were measured in the 2 groups (n = 40 for each group) using pressure-sensitive film. The mean values generated by each test group for each parameter were compared using a t test. Pearson's correlation coefficients between occlusal force, age, and time in function were examined. Occlusal force and area in the implant denture wearers were larger than those in the conventional denture wearers. No significant difference was seen in occlusal force balance between the left and right sides. The center of occlusal load in the implant denture wearers was more anterior. The maxillary denture may become more unstable in implant denture wearers. It is suggested that the stability and retention of a maxillary denture be checked and the occlusion be adjusted more frequently in the hybrid denture wearer than in the conventional denture wearer. PMID- 10453665 TI - Histologic and histomorphometric comparison of immediately placed hydroxyapatite coated and titanium plasma-sprayed implants: a pilot study in dogs. AB - The purpose of this pilot study was to make a histologic and histomorphometric comparison of hydroxyapatite-(HA) coated and titanium plasma-sprayed (TPS) root form implants that were placed in 2 mongrel dogs immediately after extraction of mandibular premolars. After 8 weeks of healing, the implant-containing segments of the mandible were removed en bloc and bone blocks including implants were sectioned. Histologic and histomorphometric analyses were performed by evaluating bone sections. The mean bone contact percentage of HA-coated implants was 61.84 +/- 7.84%, with a range of 52.09% to 75.7%, and the mean bone contact percentage of TPS implants was 51.35 +/- 12.1%, with a range of 30.1% to 70.6%. This pilot study suggests that HA-coated implants placed into fresh extraction sockets can achieve better bone contact than TPS implants, but there was evidence that the surface of the HA layer can be resorbed, so long-term stability of HA coatings in immediate implantation must be investigated. PMID- 10453666 TI - Multicenter retrospective analysis of the ITI implant system used for single tooth replacements: results of loading for 2 or more years. AB - This report involves the retrospective evaluation of ITI implants placed by a group of 12 clinicians located throughout the United States. Of the original 174 single implants placed in 129 patients reported previously, 157 were examined in 110 patients after 2 or more years (average 40.1 months). Twenty-two implants remained in the anterior and 135 implants (86%) remained in the posterior areas of the mouth, with 81 being restored with an octabutment screw-retained crown and 76 restored with a conical-abutment cemented crown (in function 2 years or longer). Occlusal screw loosening was observed in 22.2% of implants over both periods, with only 1 tooth loosening in both study periods (6 months to 2 years and > or = 2 years). Loosening of a solid conical abutment occurred in 1 additional patient, for cumulative conical abutment loosening of 5.3%. Significant radiographic bone loss was observed around 4 implants, with implant fracture noted with 3 additional implants (all mandibular first molars with hollow-screw or hollow-cylinder implant design). The survival rate at > or = 2 years was 95.5%. The data suggest that ITI implants can be a satisfactory choice for posterior single-tooth restorations. PMID- 10453667 TI - Ten-year experience in oral implant rehabilitation of cancer patients: treatment concept and proposed criteria for success. AB - Between 1988 and 1997, 18 irradiated patients (group 1, 83 implants) and 22 nonirradiated patients (group 2, 92 implants) received resection of the cancer involved mandible and floor of the mouth and subsequently underwent mandibular rehabilitation with endosseous implants. Implant-supported prostheses were placed in 26 patients, while 13 patients received implant-tissue-supported prostheses. Between 1988 and 1991, patients were restored with implant-tissue-supported prostheses (based on 2 to 4 implants). This strategy was later changed because of the development of denture-related lesions. Since 1992, group 1 patients have been restored exclusively with implant-supported prostheses on 5 to 6 implants; group 2 patients have been rehabilitated alternatively with implant-tissue supported prostheses on 4 implants. Special criteria for determining the success of implant-supported maxillofacial prostheses were developed. With a mean follow up period of 37 months, 160 implants (91%) were clinically osseointegrated. Both types of restorations provided sufficient oral rehabilitation. However, only completely implant-supported prostheses avoided soft tissue ulcers. The cumulative success rate was approximately 75% after 7 years for group 1 patients and about 86% after 10 years for group 2 patients. The success rates for implants placed after the change in strategy were approximately 86% (group 1) and 94% (group 2) after 5 years. Based on these experiences, it is suggested that irradiated patients should be restored with exclusively implant-supported prostheses, without any mucosal contact. PMID- 10453668 TI - Plasma rich in growth factors: preliminary results of use in the preparation of future sites for implants. AB - This article presents preliminary clinical evidence of the beneficial effect of the use of plasma rich in growth factors of autologous origin. The plasma is obtained from the individual patient by plasmapheresis. The macroscopic and microscopic results obtained with bone regeneration using this technique, which uses no membrane or barrier, can be observed. The incorporation of these concepts can introduce several advantages, including the enhancement and acceleration of bone regeneration and more rapid and predictable soft tissue healing. PMID- 10453669 TI - Sinus floor augmentation at the time of maxillary molar extraction: technique and report of preliminary results. AB - A technique is described for accomplishing both localized sinus augmentation and guided bone regeneration at the time of maxillary molar extraction. One hundred nine sites were treated in 92 patients. Of these, 102 procedures (94.0%) were successful and 7 (6.0%) were partially successful. Success was defined as the ability to ideally position an implant at least 10 mm in length and 4.8 mm in width without perforating the floor of the sinus or generating an implant fenestration or dehiscence. Partially successful procedures required an additional osteotome sinus lift at the time of implant placement. PMID- 10453670 TI - A comparison of 3 different endosseous nonsubmerged implants in edentulous mandibles: a clinical report. AB - The purpose of this prospective clinical study was to investigate the outcome of 3 different types of endosseous, nonsubmerged implants in the anterior part of the mandible. Fourteen older edentulous patients, 10 females and 4 males, were included. All participants received 3 different types of endosseous implants in the anterior mandible: 1 titanium plasma-sprayed cylindric implant (4 mm in diameter), 1 titanium cylindric implant with hydroxyapatite (HA) coating (4 mm in diameter), and 1 standard threaded titanium implant (3.75 mm in diameter). The 3 types of implants were originally designed to be placed in a 2-step surgical procedure. However, at this stage all implants were simultaneously provided with a temporary abutment that penetrated the mucosa. Three months later the temporary abutments were replaced by ball abutments, which were connected to an overdenture. At 12, 24, and 36 months after surgery, marginal bone resorption and Periotest values were recorded. Two patients died within the 2 first postoperative years. Five of 42 implants (11.9%) failed to osseointegrate. After 3 years, marginal bone resorption around titanium plasma-sprayed implants was significantly greater than that seen around both HA-coated and threaded titanium implants. Threaded titanium implants also had significantly better scores for marginal bone resorption than the HA-coated implants. Periotest values for HA coated cylinders were significantly lower than test values for the other implants after 3 years (P < .05). The conclusion from this investigation is that nonsubmerged implants showed impaired prognoses compared to implants placed according to the 2-stage concept. Marginal bone resorption around titanium plasma sprayed cylindric implants was clearly increased compared to the 2 other implant systems. Periotest values for HA-coated cylindric implants were superior to titanium plasma-sprayed and pure titanium implant surfaces. PMID- 10453671 TI - Augmentation grafting of the maxillary sinus and simultaneous implant placement in patients with 3 to 5 mm of residual alveolar bone height. AB - This study assessed the efficacy of augmentation grafting of the maxillary sinus with simultaneous placement of dental implants in patients with less than 5 mm of alveolar crestal bone height in the posterior maxilla prior to grafting, although the procedure has traditionally been contraindicated based on empirical data. A total of 160 hydroxyapatite-coated implants was placed into 63 grafted maxillary sinuses in 63 patients whose crestal bone height in this region ranged from 3 to 5 mm. Patients were followed for 2 to 4 years after the placement of definitive prostheses. There were no postoperative sinus complications. Following uncovering of the implants at 9 months after surgery, there was no clinical or radiographic evidence of crestal bone loss around the implants. Histologic examination of bone cores from the grafted sites revealed successful integration and a high degree of cellularity. All patients maintained stable implant prostheses during follow-up. These findings indicate that the single-step procedure is a feasible option for patients with as little as 3 mm of alveolar bone height prior to augmentation grafting, utilizing hydroxyapatite-coated implants and autogenous bone. PMID- 10453672 TI - Augmentation of the sinus floor with mandibular bone block and simultaneous implantation: a 6-year clinical investigation. AB - Between 1991 and 1995, 216 sinus-lift procedures were accomplished as part of a clinical study. The study involved placing 467 implants in the atrophic posterior maxillae of 142 female and 74 male patients. The initial bone height at the implant site was between 1 and 5 mm. The implants were supported subantrally with bone block grafts harvested from the retromolar or symphysis areas of the mandible. Perforations of the maxillary sinus membrane were observed in 51 patients; these were repaired with fibrin adhesive. The spaces remaining above the bone graft were filled with various materials. A total of 28 implants failed. All the remaining implants were deemed successfully osseointegrated, based on radiographic and clinical (including periodontal health) criteria. No patients experienced maxillary sinus complications. Clinically and radiographically, the best bone regeneration was observed in those patients in whom the surgically created space was completely grafted with autogenous bone that included a high percentage of resorption-resistant cortical bone. In those patients having bone grafts harvested from the mandibular symphysis, none of their facial profiles were adversely affected; however, some patients experienced neurosensory deficits involving the mandibular anterior incisors and adjacent alveolar mucosa. Occasionally, these symptoms persisted for up to 1 year following the procedure. PMID- 10453673 TI - A histomorphometric study of tissue interface by laminar implant test in rats. AB - Study of the implant-tissue interface is one of the fundamental issues in implantology, both odontologic and orthopedic. The characteristics of this interface will influence the success or failure of an implant. The aim of the present study was to evaluate histomorphometrically the capacity of different metals to osseointegrate employing laminar implants of zirconium, titanium, aluminum, and zirconium coated with diamond-like carbon. The experimental model herein allowed for the quantitative evaluation of the tissue-implant interface for different metals. The implants were placed in the tibiae of Wistar rats under anesthesia and allowed to remain in situ for a 30-day period. The interfaces of the zirconium and diamond-like coated zirconium implants exhibited better responses than the interface of titanium implants. Aluminum produced a local toxic effect, evidenced by osteoid formation. PMID- 10453674 TI - Le Fort I osteotomy with interpositional bone grafts and implants for rehabilitation of the severely resorbed maxilla: a 2-stage procedure. AB - A surgical procedure for the rehabilitation of severely resorbed maxillae is described. Twenty-five patients, made up of a development group of 5 and a routine group of 20, were treated with Le Fort I osteotomy using interpositional bone grafts from the iliac crest and, in a second stage, titanium implants. Altogether, 181 Branemark implants were placed, and the patients were followed for up to 5 years. The implant survival rate for the development group was 60.0% after 5 years. Life table analysis for the routine group showed a 5-year survival rate of 85.6%. Twenty-two patients received fixed prostheses and 2 received overdentures. One patient lost all implants and was rehabilitated with a prong denture. PMID- 10453675 TI - Postmortem histologic evaluation of mandibular titanium and maxillary hydroxyapatite-coated implants from 1 patient. AB - Postmortem examination of human specimens is an extremely important aspect of evaluating the relative compatibility and long-term success of endosseous implant surfaces. The bone-implant interface of 5 commercially pure titanium screw-type mandibular implants after 85 months of service and 2 hydroxyapatite- (HA) coated maxillary implants after 38 months of service were examined. All implants were stable at the time of the patient's death. The mandibular implants had an average of 65% contact with bone and the maxillary implants had an average of 47% contact. The HA coating had separated from the maxillary implants in some areas and was free within surrounding connective tissue or surrounded by invaginating sulcular epithelium. The arrangement and pattern of bone contact appeared different between HA-coated and titanium implant surfaces. PMID- 10453676 TI - Treatment of a patient with severe osteoporosis and chronic polyarthritis with fixed implant-supported prosthesis: a case report. AB - This article reports the treatment and 5-year follow-up of an 80-year-old female with a history of severe osteoporosis and chronic polyarthritis. Treatment included methotrixate disodium and acemetacin. After the last tooth was removed from the mandible, the patient was successfully treated with a fixed mandibular prosthesis supported by 6 implants placed between the mental foramina. The implants have remained osseointegrated, and peri-implant smears have been negative for bacterial colonization. Radiographic follow-up examination has revealed bone loss that is slightly greater than expected. This article focuses on the placement of implants in a patient receiving medication for chronic polyarthritis and osteoporosis. PMID- 10453678 TI - Classifying the dead: toward a history of the registration of causes of death. PMID- 10453679 TI - Cause of death as a contemporary problem. AB - Four characteristics have been identified that affect the contemporary analysis of cause of death: (1) the increasing use of cause-of-death data for public policy; (2) the growing disjuncture between cause of death and cause of poor health; (3) the problem of specifying cause of death for the elderly, who constitute a growing proportion of the population and, accordingly, of mortality; and (4) the impact of technology and medical science on the temporal comparability of mortality statistics. With regard to the first issue, statistical data in general are being used with increasing sophistication throughout society. The public is exposed to all types of data by the media,and is increasingly educated in how to interpret statistics. Government at all levels increasingly uses data to help to help identify priorities, to choose among policy options, and to evaluate outcomes of decision making. Cause-of-death data are but a specific subset of information that are being used to understand social, economic, and health issues and to make more informed private and public choices regarding these issues. Increasing use of any data set is likely to result in better data simply because greater use results in closer scrutiny of data quality with resultant emphasis on evaluation and systematic efforts to improve quality. This is happening to cause-of-death data in the United States. The National Center for Health Statistics (NCHS) initiated major efforts through two national workshops in 1989 and 1991 to improve the quality of medical certification of death through better training of physicians, medical examiners, and coroners. While there is no clear evidence that the initiatives resulting from these workshops alone resulted in data quality improvement, the indicators used by NCHS to measure quality of medical certification point to continued gradual improvement in completeness and specificity in cause-of-death data. The gap between causes of morbidity and causes of mortality is growing because advances in medical science has essentially eliminated many infectious and communicable diseases that previously caused premature death and have considerably reduced the lethality of chronic conditions that previously caused death at earlier ages. As acute conditions have moved into the background, so to speak, a variety of disabilities and impairments have moved forward as sources of illness,pain, and diminished quality of life. As a consequence, the previously clear etching in mortality statistics of a society's health has become blurred by advances in medicine, healthcare, and prevention. Mortality data continue to have value per set and are important to social and medical knowledge because of their availability as a uniquely continuous data set. However, increasingly, a complete and contemporary profile of the health of a society requires data on both morbidity and mortality as complimentary indicators of health and social development. Not only do we need to know of what causes people to die, but also what health conditions aggravate and diminish their lives and, at an aggregate level, that conditions impose the greatest health costs on the community. Increasingly, these are debilitating conditions of the elderly that may diminish life but may not cause death. Medical description of the cause of death of an elderly person has probably never been easy; however, the contemporary extension of life has made the problem more visible and substantial than in earlier times. It is likely that the relatively few octogenarians of centuries past were also afflicted by concurrent medical conditions, and that even younger persons were prematurely afflicted by conditions that are now identified with advanced age. For these population groups, the description of cause of death, as a single and singular initiating event, is problematic. PMID- 10453680 TI - The development of reporting systems for causes of deaths in Denmark. PMID- 10453681 TI - Recording the epidemiologic transition in Germany, 1816-1934. PMID- 10453682 TI - Reporting causes of death in Sweden, 1750-1950. PMID- 10453683 TI - National statistics on the causes of death in nineteenth-century Bavaria. PMID- 10453684 TI - Premises, premises: comments on the comparability of classifications. PMID- 10453685 TI - Medical causes of death in preindustrial Europe: some historiographical considerations. PMID- 10453686 TI - Nosology, mortality, and disease theory in the eighteenth century. PMID- 10453687 TI - Problems of classifying death in nineteenth-century Japan. PMID- 10453688 TI - Parochial burial registers: the case of Texas in the nineteenth century. PMID- 10453689 TI - The measurement of maternal mortality. PMID- 10453690 TI - Missouri State Medical Association membership directory. PMID- 10453691 TI - Macrophage migration inhibitory factor: cytokine, hormone, or enzyme? PMID- 10453692 TI - Voltage-dependent calcium channels: from structure to function. PMID- 10453693 TI - Aging of the male endocrine system. PMID- 10453694 TI - Effect of age on thymic development, T cell immunity, and helper T cell function. PMID- 10453695 TI - Aging and chromosomal instability. PMID- 10453696 TI - [XXIV Congress of the Spanish Society of Anesthesiology, Resuscitation and Pain Therapy. Sevilla, Spain, 11-15 May 1999. Abstracts]. PMID- 10453697 TI - Cholera, 1998. PMID- 10453699 TI - Gold Medal Award for Life Achievement in the Science of Psychology. PMID- 10453700 TI - Gold Medal Award for Enduring Contributions to Psychology in the Public Interest. PMID- 10453701 TI - Gold Medal Award for Life Achievement in the Practice of Psychology. PMID- 10453702 TI - Gold Medal Award for Life Achievement in the Application of Psychology. PMID- 10453703 TI - Award For Distinguished Teaching in Psychology. PMID- 10453704 TI - Guidelines for psychological evaluations in child protection matters. Committee on Professional Practice and Standards, APA Board of Professional Affairs. PMID- 10453705 TI - HIV and infant feeding: to breastfeed or not to breastfeed: the dilemma of competing risks. Part 1. AB - The discovery of the human immunodeficiency virus (HIV) in breastmilk in 1985, and subsequent research, supports the hypothesis that breastfeeding provides a route of transmission to the nursing baby. Various routes of infection and relative rates of transmission have been studied in many parts of the world, leading to the blanket guideline that babies of HIV-infected mothers should not be breastfed, if a safe alternative can be provided. However, due to the limits inherent in various studies and various testing methods, the exact frequency of breastmilk transmission of HIV during the course of lactation remains unknown, and the conclusions drawn are thus conflicting and confusing. Replacement feeding of young babies with non-human milks and other foods may be hazardous in poverty stricken populations in Africa and elsewhere, and still more research suggests that there are several properties in human milk that may provide specific protection to the baby of an infected mother. The possibility of providing the mother's own treated expressed breastmilk to the baby at risk of HIV infection via breastfeeding is an alternative which has yet to be fully explored and ways that this could be accomplished are examined. Those of us working with mothers and babies need more information before we can assist mothers living with HIV to make truly informed decisions about the safest way to feed their babies. Topics requiring urgent further attention are outlined. PMID- 10453706 TI - HIV and infant feeding: a policy statement developed collaboratively by UNAIDS, WHO and UNICEF, 1997. PMID- 10453707 TI - Promoting breastfeeding in the neonatal intensive care unit. AB - The technology and separation of the mother and baby in the neonatal unit make bonding difficult and create barriers to breastfeeding. This pilot study aimed to define those nursing interventions that promote breastfeeding. The neonatal nurse plays a vital role in the influence of the breastfeeding outcome. Forty-one preterm babies had their breastfeeding activities, (including kangaroo care), recorded daily. The specific phases in preterm infants of 32 weeks or more were identified as 1. stabilisation and assessing ability, 2. controlled challenge, 3. progression and regression and 4. stability of feeding activity. This study has increased the awareness among the neonatal nurses involved as to the nursing practices that promote breastfeeding and has provided a basis from which nurses can promote breastfeeding to mothers of preterm babies. PMID- 10453708 TI - An analysis of breastfeeding initiation in Tasmania by demographic and socioeconomic factors for the period 1981-1995. AB - Breastfeeding initiation and duration trend data is an important component of infant health and nutrition monitoring and surveillance systems. It is also an important basis for identifying breastfeeding promotion needs, prioritising target groups and strategies and in evaluating the effectiveness of breastfeeding promotion efforts. A collation and analysis of available Tasmanian breastfeeding initiation data was undertaken in order to help direct statewide breastfeeding promotion planning and evaluation and to begin the development of a minimum data set for statewide breastfeeding monitoring and surveillance. Perinatally collected breastfeeding initiation data (breastfeeding at discharge from hospital) was analysed against available demographic and socioeconomic variables and compared with initiation rates from interstate Australian populations. Data suggest that breastfeeding initiation rates in Tasmania have been static over the 15 years to 1995 (a three percent increase from 1981 to 1995) and that the associations between breastfeeding initiation and occupational classification, maternal age and parity in Tasmania are consistent with those reported elsewere. Whilst caution needs to be applied to between-population comparisons, breastfeeding initiation rates in Tasmania appear to be considerably lower than that reported in other states. This suggests relatively greater efforts are required to influence breastfeeding initiation. Further research is required to investigate reasons for this difference. PMID- 10453709 TI - Older babies who bite at the breast. PMID- 10453710 TI - Sizing up the new arthritis drugs. PMID- 10453712 TI - Getting to the cause of sleep problems. PMID- 10453711 TI - Can cholesterol lowering be too aggressive? PMID- 10453713 TI - Diagnosing and treating rectal problems. PMID- 10453714 TI - A brief history of hypertension. PMID- 10453715 TI - I have congestive heart failure. Should I try coenzyme Q-10? PMID- 10453716 TI - My carpal tunnel surgery was unsuccessful. What now? PMID- 10453717 TI - I have mild hypertension and my doctor recommended I eat bananas and drink orange juice. Why? PMID- 10453718 TI - Anticancer effect of 9-nitrocamptothecin liposome aerosol on human cancer xenografts in nude mice. AB - PURPOSE: To test the anticancer properties of the water-insoluble derivative of camptothecin, 9-nitrocamptothecin (9-NC) against human breast, colon and lung cancer xenografts in nude mice when administered in liposome aerosol. METHODS: The drug was formulated with dilauroylphosphatidylcholine and nebulized in a particle size of 1.6 microm +/- 2.0 mass median diameter to deliver doses of usually less than 200 microg/kg daily, 5 days per week. 9-NC liposome aerosols were generated with a Aerotech II nebulizer (CIS-USA) flowing at 101/min from a compressed air source and delivered to mice in sealed plastic cages or in a nose only exposure chamber. RESULTS: Tumor growth was greatly reduced or tumors were undetectable after several weeks of treatment. Colon tumor was least responsive. 9-NC was better than the parent compound, camptothecin, also water-insoluble, tested by aerosol in a similar liposomal preparation. Equivalent doses of 9-NC liposome preparations administered by mouth were substantially without effect while there was some effect, but limited, of the liposome preparation given intramuscularly. CONCLUSIONS: 9-NC liposome aerosol was strikingly effective in the treatment of three human cancer xenografts growing subcutaneously over the thorax in nude mice at doses much smaller than those traditionally used in mice administered by other routes. PMID- 10453719 TI - Distribution of camptothecin after delivery as a liposome aerosol or following intramuscular injection in mice. AB - PURPOSE: The plant alkaloid camptothecin (CPT) has shown significant antitumor activity against a wide variety of human tumors xenografted in nude mice. In previous studies we have found that administration of dilauroylphosphatidylcholine (DLPC) liposome aerosols containing 9 nitrocamptothecin (9-NC) inhibits the growth of human breast, colon and lung cancer xenografts. The purpose of this study was to analyze the pharmacokinetics and tissue distribution of inhaled CPT formulated in DLPC liposomes. METHODS: C57BL/6 mice with subcutaneous Lewis lung carcinoma, Swiss nu/nu mice with human lung carcinoma xenografts and BALB/c mice without tumors were used for pharmacokinetic studies of CPT administered as a liposome aerosol and BALB/c mice were given CPT intramuscularly. RESULTS: After 30 min inhalation of CPT liposome aerosol, drug was deposited in the lungs (310 ng/g) and was followed promptly by the appearance of high concentrations in the liver (192 ng/g) and with lesser amounts appearing in other organs. Drug concentration in the brain was 61 ng/g. After intramuscular injection of CPT dissolved in DMSO, drug was released from the site of injection very slowly and accumulated mainly in the liver (136 ng/g). Only trace amounts appeared in the lungs (2-4 ng/g). These results demonstrate a prompt pulmonary and later systemic distribution of CPT following liposome aerosol administration. CONCLUSIONS: The substantial concentrations of CPT in lungs and other organs following inhalation of liposome aerosol suggest the possible benefit of it and of its more active derivative, 9-NC, in the treatment of lung, liver, kidney and brain cancer in humans. PMID- 10453720 TI - Cisplatin efflux, binding and intracellular pH in the HTB56 human lung adenocarcinoma cell line and the E-8/0.7 cisplatin-resistant variant. AB - PURPOSE: Many cell lines resistant to cisplatin (DDP) have reduced DDP accumulation. We postulated that reduced accumulation of DDP in resistant cells might be due to decreased intracellular DDP binding, leading to increased passive efflux. METHODS: The total cellular ([T-DDP]), intracellular ultrafiltrable ([F DDP]) and precipitable cellular bound ([B-DDP]) DDP concentrations were all compared in the HTB56 human lung adenocarcinoma cell line and its E-8/0.7 variant that has acquired DDP resistance. Cells were exposed to 509 microM DDP for 20 min. Ultrafiltration with a 500 molecular weight cut-off separated cellular free from bound cisplatin. Fragmentation by sonication and microcentrifugal spinning precipitated cellular bound cisplatin. Flow cytometry was used to measure the intracellular pH (pHi) of the HTB56 cell line, the E-8/0.7 cell line, as well as of the OV2008 cell line and its C13 resistant variant. The DNA-bound DDP and protein-bound DDP ([P-DDP]) were also compared when equal [T-DDP] was achieved for both sensitive and resistant cells by exposing them for 1 h to two pairs of DDP concentrations, i.e. 509 vs 911 microM DDP, and 111 vs 666 microM DDP, respectively. Platinum was assayed by flameless atomic absorption spectrophotometry. RESULTS: At time 0 (end of cisplatin exposure), [T-DDP] and [B DDP] were significantly higher in the sensitive HTB56 parent cell line (P < 0.02 and P < 0.001, respectively), whereas [F-DDP] did not differ significantly (P = 0.62). Two distinct phases of T-DDP efflux were observed. In the first 10 s after DDP exposure, the rate constant for resistant cells (KR1) was 0.17 s(-1), whereas that for sensitive cells (KS1) was 0.14 s(-1). From 10 s to 50 s, however, KR2 and KS2 became 0.005 s(-1) and 0.004 s, respectively. [T-DDP] remained lower in resistant cells than in sensitive cells at 10, 30 and 50 s (all P < 0.0001). For 1 h drug exposure to 509 vs 911 microM cisplatin concentrations designed to give comparable [T-DDP] in the sensitive and resistant cell lines, only [DNA-bound DDP] was found to be significantly higher in sensitive cells (P = 0.002), whereas both [F-DDP] and [P-DDP] did not differ significantly (P = 0.18, P = 0.75, respectively). On the other hand, there were no significant differences found in [F-DDP], [P-DDP] and [DNA-bound DDP] between the two cell lines when 111 vs 666 microM DDP was used. Flow cytometry data indicated that the pHi was significantly higher in the E-8/0.7 (P < 0.0186) and C13 (P < 0.0169) resistant variants than in the sensitive parent cell lines. CONCLUSIONS: DDP binds more slowly in resistant than in sensitive lung cancer cells, despite comparable amounts of free drug. Early efflux is higher in the resistant variant. Differences between the lines with respect to DNA binding may be DDP concentration-dependent. We speculate that the reduced early binding and increased early efflux in the resistant line may be related to the higher pH in this line. A higher pH is supposed to favor production of neutral hydroxyl metabolites rather than charged aquated metabolites, and these neutral metabolites would be expected to react less readily with intracellular molecules and to efflux more readily across cell membranes. Since we have previously documented a threefold increase in glucose utilization and lactate production in the DDP-resistant variants of the human HTB56 lung cancer cell lines, and this increased lactate production would have been expected to reduce the intracellular pH instead of raising it, it is possible that our alkaline-resistant cells have a higher Na+/H+ exchanger activity which would protect them from intracellular acidification. PMID- 10453721 TI - Vincristine pharmacokinetics after repetitive dosing in children. AB - PURPOSE: We studied vincristine disposition after 169 weekly i.v. bolus injections in 32 children with acute lymphoblastic leukemia, non-Hodgkin lymphoma, or Wilms' tumor. The aim of the study was to determine intrapatient and interpatient variability in vincristine disposition and demographic, clinical, and biochemical characteristics influencing this variability. METHODS: Vincristine plasma concentrations were measured by a high-performance liquid chromatography assay with electrochemical detection. A limited sampling strategy was used based on a bayesian parameter estimation algorithm that is part of the ADAPT II software package. A two-compartment, first-order model was fitted to the data, and pharmacokinetic parameters were calculated from the model using the ADAPT II software. For statistical analysis, analysis of variance (ANOVA), t test, simple and multiple regression analysis, and non-parametric or robust equivalents were used. RESULTS: Results showed a large intrapatient and interpatient variability in distribution half-life, elimination half-life, total body clearance, apparent volume of distribution at steady state, and area under the concentration-time curve. Intrapatient variability was significantly smaller than interpatient variability for all these parameters except distribution half life. The diagnosis or treatment protocol turned out to be the most predictive characteristic; leukemia and non-Hodgkin lymphoma patients had a significantly higher total body clearance than Wilms' tumor patients. CONCLUSIONS: We conclude that both intrapatient and interpatient variability in vincristine pharmacokinetics is large in pediatric cancer patients and that variability, although significantly influenced by diagnosis, largely remains unpredictable. PMID- 10453722 TI - Phase II study of a radiotherapy/etoposide combination for patients with newly malignant gliomas. AB - PURPOSE: Etoposide, a semisynthetic derivative of podophyllotoxine, is a topoisomerase II inhibitor. This drug is currently used in several types of human cancer. The aim of this study was to evaluate the efficacity and tolerance of a near-concurrent association of radiotherapy and etoposide for newly malignant gliomas. METHODS: From May 1995 to December 1996, 30 malignant glioma patients were included in this phase II study; 16 patients underwent surgical tumor resection, and a stereotactic biopsy was performed in 14 patients. Standard cranial irradiation and six courses of etoposide (100 mg/m2, x days 1-3) were administered. The first course of etoposide was administered on days 1-3 of radiotherapy and was resumed in the week following the end of radiotherapy. Treatment was consolidated by further courses of etoposide every 4 weeks. RESULTS: Only 26 patients could be evaluated for the purpose of our study. The median age was 60.1 years, and the median Karnofsky performance score (KPS) was 80.2. The rate of objective response for evaluable patients was 34.6%, and four complete responses (CR) and five partial responses (PR) were noted. The median survival (MST) was 12 months, and the average overall survival was 12.5 months. Hematological toxicity was mild, and grade 3 or 4 neutropenia (white blood cell count < 1500/ml) was noted in three patients, without any sepsis or bleeding. CONCLUSIONS: The results obtained in this study are comparable to the best reported results on the combination of radiotherapy and nitrosoureas. The near concurrent combination of radiotherapy and etoposide seems to be effective and well tolerated in the treatment of newly malignant gliomas. PMID- 10453723 TI - Failure of taxol-based combination chemotherapy for malignant glioma cannot be overcome by G2/M checkpoint abrogators or altering the p53 status. AB - In an effort to develop more effective forms of adjuvant chemotherapy for malignant brain tumors, we sought to develop a taxol-based combination chemotherapy regimen for glioma cell lines in vitro. Here, we report that coexposure of LN-229 or T98G glioma cells to taxol and either doxorubicin, camptothecin, cytarabine, or VM26 resulted in antagonistic effects rather than additive or synergistic cytotoxicity. There were no interactions of taxol with the effects of carmustine (BCNU) or vincristine. Antagonism was more prominent in cytotoxic cell death assays than in clonogenic cell death assays and was not overcome by G2/M checkpoint abrogators such as caffeine or pentoxyfilline. Ectopic expression of mutant and wild-type p53val135 attenuated taxol cytotoxicity in both T98G cells, which are mutant for p53, and LN-229 cells, which exhibit functional wild-type p53 activity. Interestingly, wild-type p53val135 abrogated the taxol-imposed G2/M arrest in both cell lines. However, wild-type p53val135 did not promote glioma cell killing by combinations of taxol and any of the other drugs. Further, an analysis of a panel of 12 human glioma cell lines revealed no relationship between genetic or functional p53 status and taxol sensitivity. In summary, combination either with other chemotherapy drugs, with abrogators of the G2/M checkpoint, with wild-type p53 gene transfer was not a promising approach for a taxol-based combination chemotherapy regimen in malignant glioma. PMID- 10453724 TI - Phase I/pharmacokinetic trial of the novel thioxanthone SR233377 (WIN33377) on a 5-day schedule. AB - PURPOSE: SR233377 (WIN33377) is a novel 4-aminomethyl thioxanthone derivative with promising preclinical activity against solid tumors at doses substantially below the MTD. We performed a phase I trial to determine a suitable phase II dose of SR233377 when administered as a 2-h intravenous infusion for five consecutive days. METHODS: A group of 25 patients with a range of solid tumor diagnoses and good performance status received SR233377 at eight dose levels ranging from 4.8 mg/m2 per day to 74.7 mg/m2 per day. Cycles were repeated every 35 days and patients were evaluated for response following two cycles of treatment. Doses were escalated in cohorts of three using a modified Fibonacci scheme. Pharmacokinetic sampling was performed during the first cycle in all patients. RESULTS: Toxicities of SR233377 on this schedule included neutropenia, fever, nausea, and dyspnea but all were mild and not dose-limiting. Asymptomatic prolongation of the corrected QT (QTc) interval during infusion in all patients monitored at the 74.7 mg/m2 dose level prompted closure of the study. QT lengthening correlated with increasing plasma concentrations of SR233377. SR233377 Cmax values increased linearly with dose, but substantial interpatient variability in SR233377 AUC, clearance, and half-life was noted. There was no evidence of drug accumulation when day 1 and day 5 AUC and Cmax values were compared. Seven patients displayed tumor growth inhibition lasting for 4 months or more. CONCLUSIONS: We conclude that SR233377 administered on a 5-day schedule is associated with tolerable clinical symptoms and some activity against a range of solid tumors but dosing is limited by QTc prolongation, a condition that predisposes to ventricular arrhythmias. Phase II development on this schedule is not recommended based on the occurrence of this concentration-dependent effect. Further investigation of alternative schedules of administration and of SR233377 analogues is warranted. PMID- 10453725 TI - Characterization of MGI 114 (HMAF) histiospecific toxicity in human tumor cell lines. AB - PURPOSE: The acylfulvenes are a class of antitumor agents derived from the fungal toxin illudin S. One acylfulvene derivative, MGI 114 (HMAF), demonstrates marked efficacy in xenograft carcinoma models when compared to the parent acylfulvene or related illudin compounds. The maximum tolerated dose (MTD) of the two analogs in animals, however, is similar. To help elucidate the basis of the increased therapeutic efficacy of MGI 114, we determined the in vitro cytotoxicity, cellular accumulation and DNA incorporation of this drug and compared the results with those from the parent acylfulvene analog. METHODS: The cytotoxicity of acylfulvene analogs was tested in vitro against a variety of tumor cell lines. Radiolabeled MGI 114 was used for cellular accumulation and DNA incorporation studies. RESULTS: MGI 114 retained relative histiospecific toxicity towards myeloid leukemia and various carcinoma cell lines previously noted with the parent acylfulvene compound. Markedly fewer intracellular molecules of MGI 114 were required to kill human tumor cells in vitro as compared to the parent acylfulvene, indicating that MGI 114 was markedly more toxic on a cellular level. At equitoxic concentrations, however, the incorporation of MGI 114 into genomic tumor cell DNA was equivalent to that of acylfulvene. Analysis of cellular accumulation of MGI 114 into tumor cells revealed a lower Vmax for tumor cells, and a markedly lower Vd for diffusion accumulation as compared to acylfulvene. CONCLUSIONS: The addition of a single methylhydroxyl group to acylfulvene to produce MGI 114 results in a marked increase in cytotoxicity in vitro towards tumor cells as demonstrated by the reduction in IC50 values. There was a corresponding decrease in the number of intracellular molecules of MGI 114 required to kill tumor cells, but no quantitative alteration in covalent binding of the drugs to DNA at equitoxic concentrations. This indicates that cellular metabolism plays a role in the in vitro cytotoxicity of MGI 114. The equivalent incorporation into genomic DNA at equitoxic doses suggests that DNA damage produced by acylfulvene and MGI 114 is equivalent in regard to cellular toxicity and ability to repair DNA. This increased cellular toxicity, together with the decrease in diffusion rate, may explain the increased therapeutic efficacy of MGI 114 as compared to the parent acylfulvene analog. PMID- 10453726 TI - Cremophor reduces paclitaxel penetration into bladder wall during intravesical treatment. AB - PURPOSE: We have previously shown that paclitaxel, when dissolved in water and instilled into the bladder, readily penetrates the urothelium. The FDA-approved formulation uses Cremophor and ethanol to dissolve paclitaxel. In the present study, the effects of this solvent system on the urine, bladder tissue, and plasma pharmacokinetics of intravesical paclitaxel were evaluated. METHODS: Plasma, urine, and tissue pharmacokinetics were determined in five dogs treated for 120 min with paclitaxel (500 microg per 20 ml of 0.22% w/v Cremophor and 0.21% v/v ethanol) by intravesical instillation. Equilibrium dialysis was used to determine the free fraction of paclitaxel and the presence of Cremophor micelles was verified using a fluorescent probe method. RESULTS: The average bladder tissue concentration was > 1600-fold higher than the plasma concentration. Comparison of the results for paclitaxel dissolved in Cremophor/ethanol with our previous results of paclitaxel dissolved in water (500 microg per 20 ml) indicates that Cremophor/ethanol decreased the paclitaxel partition across the urothelium and reduced the average bladder tissue concentration by 75%, but did not alter the rate of paclitaxel penetration across the bladder wall, the urine pharmacokinetics or the plasma pharmacokinetics of paclitaxel. For Cremophor, the urine concentrations during the 120-min treatment ranged from 0.12% to 0.22%, and the concentration in bladder tissue from 0.00004% to 0.0009%. The threshold Cremophor concentration for micelle formation was 0.008%. We found that ethanol at concentrations up to 1% and Cremophor at concentrations below 0.01% did not alter the free fraction of paclitaxel, whereas Cremophor at higher concentrations, i.e. 0.065% and 0.25%, significantly reduced the free fraction by two- to six-fold, respectively. These results indicate that during intravesical instillation of the FDA-approved paclitaxel formulation, the concentration of Cremophor in urine was sufficient to form micelles, resulting in sequestration of paclitaxel into micelles, reduction in the free fraction of paclitaxel and consequently a reduction in paclitaxel penetration across the urothelium. In contrast, the Cremophor concentrations in bladder tissue were inadequate to form micelles and thus did not alter the drug penetration through the bladder tissue. CONCLUSIONS: We conclude that intravesical paclitaxel treatment using the FDA approved formulation provides a significant bladder tissue targeting advantage, although the advantage is lower than when paclitaxel is dissolved in water. PMID- 10453727 TI - Enhancement of cisplatin cytotoxicity by terbium in cisplatin-resistant MDA/CH human breast cancer cells. AB - PURPOSE: The development of cisplatin resistance is a major problem in the treatment of cancer patients with cisplatin chemotherapy. The membrane binding of terbium (Tb3+) has been shown to increase the cellular accumulation of cisplatin in breast cancer cells. Therefore, the ability of Tb3- to modulate the cytotoxicity of cisplatin was investigated in cisplatin-sensitive (MDA) and cisplatin-resistant (MDA/CH) MDA-MB-231 human breast cancer cells. METHODS: The cytotoxic parameters of cisplatin were determined using live cell microfluorometry and median effect analysis. RESULTS: MDA/CH cells (IC50 = 142 +/ 9 microM) were found to be approximately 3.3-fold more resistant to cisplatin than MDA cells (IC50 = 43.5 +/- 3.0 microM). In both cell lines, the IC50 value for cisplatin was reduced two-fold in the presence of 80 microM Tb3+, thus indicating that the cytotoxicity of cisplatin is increased by Tb3+. The cytotoxic activity of cisplatin alone was observed to be 5.7 and 1.6 times more potent than that of Tb3+ alone in MDA and MDA/CH cells, respectively. Combination index analyses revealed that the interaction between cisplatin and Tb3+ was only synergistic at very low indices of cell death in MDA cells. However, in MDA/CH cells, the two drugs were synergistic up to intermediate levels of cell death. CONCLUSIONS: Our results suggest that the enhancement of cisplatin cytotoxicity by Tb3- is more effective in cisplatin-resistant MDA/CH cells than in cisplatin sensitive MDA cells. Therefore, terbium is potentially useful in cisplatin combination therapy for breast cancer patients, especially for those patients who have developed resistance to the drug. PMID- 10453728 TI - A phase II study of the effectiveness of docetaxel (Taxotere) in women with advanced breast cancer previously treated with polychemotherapy. Hellenic Cooperative Interhospital Group in Oncology. AB - PURPOSE: The aim was to study the effectiveness of docetaxel (Taxotere) in patients with advanced breast cancer treated previously with polychemotherapy. PATIENTS AND METHODS: Forty-nine patients received docetaxel (100 mg/m2; 1-h i.v. infusion) and corticosteroid premedication. Forty-one patients who had received previous anthracycline treatment were divided into anthracycline-refractory and anthracycline-resistant (early and late) groups. RESULTS: Of 45 evaluable patients, 66.7% had a partial response (PR) and 2.2% a complete response (CR), giving an overall response rate (ORR) of 68.9%. The ORR in anthracycline refractory patients was 60% versus 82.6% in anthracycline-resistant patients; the difference was not significant. The ORR in early-resistance patients was 62.5% versus 93.4% in late-resistance patients (0.05 < P < 0.1). The median response duration and overall survival was 8 months (range, 4-23 + months) and 11.5 months (range, 4-31 + months), respectively, in 39 patients treated previously for metastatic disease. For 295 courses, grade 3/4 neutropenia developed in 28.6% of patients (12.5% of courses) and was febrile in 26.5% of patients (6.1% of courses), including one septic death. Hypersensitivity reactions (HSR) developed in 16.3% of patients, and fluid retention developed in 34.7% of patients (11.9% of courses). CONCLUSIONS: Docetaxel is an active second-line drug in advanced breast cancer. The time of relapse after cessation of anthracycline treatment may be a significant prognostic factor. PMID- 10453729 TI - A phase II/pharmacokinetic trial of high-dose progesterone in combination with paclitaxel. AB - PURPOSE: The purpose of this study was to investigate the effect of high-dose progesterone, an inhibitor of P glycoprotein, on the pharmacokinetics and toxicity of paclitaxel. PATIENTS AND METHODS: A total of 29 patients with various tumors were treated with single-agent paclitaxel (125 mg/m2 administered over 3 h once every 3 weeks) until progression of disease, at which point high-dose progesterone (3 g administered i.v. over 24 h) was added to the paclitaxel treatment program in 20 patients (13 women, 7 men). Pharmacokinetic studies of paclitaxel administered alone and with progesterone were performed in eight patients. RESULTS: The pharmacokinetic parameters of paclitaxel were highly variable. High-dose progesterone increased the peak plasma levels (3.00 +/- 0.94 vs. 4.15 +/- 1.63 microM; P = 0.029; mean +/- SD) and the area under the curve (AUC; 14.3 +/- 4.75 vs. 17.3 +/- 5.59 microM x h; P = 0.006) of paclitaxel. The absolute neutrophil and platelet nadir counts did not differ significantly between the paclitaxel and the combined treatment cycles. Three of the 20 patients documented to have progressive disease on paclitaxel alone had partial responses when high-dose progesterone was added to the paclitaxel regimen. CONCLUSION: Progesterone had a statistically significant impact on the pharmacokinetics of paclitaxel. The addition of high-dose progesterone to paclitaxel is feasible, but the small number of patients prevents conclusions being drawn about the clinical efficacy of combined progesterone and paclitaxel. PMID- 10453730 TI - Hereditary spastic paraplegia: mitochondrial metalloproteases of yeast. AB - Hereditary spastic paraplegia (HSP) is a genetically heterogenous group of inherited neurodegenerative disorders. Recently, an autosomal recessive form of HSP was mapped to 16q24.3, and subsequently the defective gene associated to HSP was identified and designated SPG7. The SPG7 gene product was predicted to encode a protein of 795 amino acids, and is called paraplegin. Paraplegin is highly homologous to a class of well studied yeast ATP-dependent zinc metalloproteases, which show 55%, 55% and 52% identity, respectively, to Afg3p, Rca1p and Yme1p. Mutation of either Afg3p, Rca1p or Yme1p in yeast results in pleiotropic effects with regard to growth, respiration and, particularly, in the assembly and/or degradation of more than one mitochondrial protein complex. Taking into account the homology of paraplegin to these yeast ATP-dependent zinc metalloproteases and what is known about their function, allows us to speculate as to what function paraplegin plays in normal individuals. PMID- 10453731 TI - Evaluation of the Best disease gene in patients with age-related macular degeneration and other maculopathies. AB - Vitelliform macular dystrophy (VMD2, Best disease, MIM153700) is an early onset, autosomal, dominant macular degeneration characterized by the deposition of lipofuscin-like material within and below the retinal pigment epithelium (RPE); it is associated with degeneration of the RPE and overlying photoreceptors. Recently, we cloned the gene bestrophin, which is responsible for the disease, and identified a number of causative mutations in families with VMD2. Here, we report that the analysis of bestrophin in a collection of 259 age-related macular degeneration (AMD) patients provides evidence that mutations in the Best disease gene do not play a significant role in the predisposition of individuals to AMD. However, our results suggest that, in addition to Best disease, mutations within the bestrophin gene could be responsible for other forms of maculopathy with phenotypic characteristics similar to Best disease and for other diseases not included in the VMD category. PMID- 10453732 TI - NME6: a new member of the nm23/nucleoside diphosphate kinase gene family located on human chromosome 3p21.3. AB - The NME (nm23/nucleoside diphosphate kinase) gene family in human is involved in the phosphorylation of nucleoside diphosphates and a variety of regulatory phenomena associated with development, oncogenic transformation, and metastasis. Here we report the cDNA sequence for a sixth member of this family, NME6. The cDNA sequence predicts a 186-residue protein that includes the characteristic active site motif of a nucleoside diphosphate (NDP) kinase, as well as the other residues previously identified as crucial for nucleotide binding and catalysis. The NME6 protein sequence is only 34-41% identical to the five previously reported human NME proteins, and is similarly related to prokaryotic and primitive eukaryotic NDP kinases. Compared to typical proteins of this family such as NME1 and NME2, NME6 has three additional residues located in the Kpn loop, and a 22-residue extension at the COOH-terminal. Using radiation hybrid mapping, the NME6 gene was localized to chromosome 3p21.3. The 1.3-kb transcript of NME6 is expressed at a moderately low level in many human tissues, and is most abundant in kidney, prostate, ovary, intestine, and spleen. Homologous cDNAs were also cloned and sequenced for rat and mouse. The sequence of the first 171 residues of the mouse homologue (Nm23-M6) is 94% identical to the deduced human NME6 protein. PMID- 10453733 TI - A mitochondrial cytochrome b mutation but no mutations of nuclearly encoded subunits in ubiquinol cytochrome c reductase (complex III) deficiency. AB - Ubiquinol cytochrome c reductase (complex III) deficiency represents a clinically heterogeneous group of mitochondrial respiratory chain disorders that can theoretically be subject to either a nuclear or a mitochondrial mode of inheritance. In an attempt to elucidate the molecular bases of the disease, we first determined the nucleotide sequence of three unknown subunits (9.5 kDa, 7.2 kDa, 6.4 kDa) by cyberscreening of human expressed sequence tag data bases and sequenced the 11 cDNA subunits encoding complex III in five patients with isolated complex III deficiency. No mutation in the nuclearly encoded complex III subunits was observed, but a mutation in the cd2 helix of the mitochondrial (mt) cytochrome b gene was found to alter the conformation of the bc1 complex in one patient with severe hypertrophic cardiomyopathy. The present study is highly relevant to genetic counseling as the absence of mtDNA mutations in all but one patient in our series strongly supports autosomal rather than maternal inheritance in the majority of patients with complex III deficiency. PMID- 10453734 TI - Gene expression patterns in cell lines from patients with 18q- syndrome. AB - Some studies have suggested that for trisomies, some genes are expressed far in excess of the expected 150% level and that this "dysregulation" is one of the mechanisms for the pathogenesis of trisomies. In an attempt to generalize this result to a monosomy, we examined mRNA isolated from lymphoblastoid cell lines derived from patients with 18q- syndrome, a deletion syndrome involving loss of the distal long arm of chromosome 18. Expression levels of ten chromosome 18 genes were compared between cell lines from eight patients with 18q- syndrome and four diploid controls. Gene expression was investigated by a quantitative reverse transcription polymerase chain reaction (RT-PCR) method. With the exception of the transcription factor NFATC1, which shows a tendency towards gene dosage compensation (the expression pattern correlates with IgA deficiency), all of the other genes were expressed at a level proportional to their gene copy number. This was true regardless of mRNA abundance or different patterns of gene expression (ubiquitous versus tissue-specific gene expression). These results indicate that, unlike dysregulated gene expression apparent in some trisomies, this monosomic syndrome is largely due to consequences of reduced gene expression. PMID- 10453735 TI - Hyperinsulinism-hyperammonemia syndrome caused by mutant glutamate dehydrogenase accompanied by novel enzyme kinetics. AB - Hyperinsulinism-hyperammonemia syndrome (HHS) is a recently identified genetic disorder characterized by hyperinsulinemic hypoglycemia with concomitant hyperammonemia. In patients with HHS, activating mutations in the glutamate dehydrogenase (GDH) gene have been identified. GDH is a key enzyme linking glutamate metabolism with the Krebs cycle and catalyzes the conversion of glutamate to alpha-ketoglutarate. The activity of GDH is controlled by allosteric inhibition by GTP and, so far, all the mutations of HHS patients have been located within the GTP-binding site. Characteristically, GDH from these individuals have therefore normal basal activity in conjunction with a loss of GTP inhibition. In this study, however, we have identified a novel variant GDH in a patient with a more severe form of HHS. The mutation is located outside the GTP binding site and the patient's GDH shows consistently higher activity, even in the absence of allosteric effectors. These results further support the hypothesis that the activating mutation of GDH is the cause of HHS. The mechanism leading to the activation of GDH, however, is not always related to the loss of GTP inhibition as was originally suggested. PMID- 10453736 TI - Genetic anticipation in Portuguese kindreds with familial amyloidotic polyneuropathy is unlikely to be caused by triplet repeat expansions. AB - Familial amyloidotic polyneuropathy (FAP) is a lethal autosomal dominant type of amyloidosis resulting from the deposition of transthyretin (ATTR) variants in the peripheral and autonomic nervous systems. ATTR V30M-associated FAP exhibits marked genetic anticipation in some families, with clinical symptoms developing at an earlier age in successive generations. The genetic basis of this phenomenon in FAP is unknown. Anticipation has been associated with the dynamic expansion of trinucleotide repeats in several neurodegenerative disorders, such as Huntington disease, myotonic dystrophy, and fragile X syndrome. We have used the repeat expansion detection (RED) assay to screen affected members of Portuguese FAP kindreds for expansion of any of the ten possible trinucleotide repeats. Nine generational pairs with differences in their age of onset greater than 12 years and a control pair with identical ages of onset were tested. No major differences were found in the lengths of the ten trinucleotide repeats analyzed. The distribution of the maximal repeat sizes was consistent with reported studies in unrelated individuals with no known genetic disease. The present data do not support a role for trinucleotide repeat expansions as the molecular mechanism underlying anticipation in Portuguese FAP. PMID- 10453737 TI - Ancestral origin of variation in the triosephosphate isomerase gene promoter. AB - A high frequency of nucleotide substitutions -5A/G, -8G/A, -24T/G in the triosephosphate isomerase (TPI) gene promoter has been demonstrated in African Americans. The biological significance of these promoter variants, two of which, 8G/A and -24T/G, occur within regulatory elements essential for transcription, is controversial. The geographical distribution and frequency of allelic variation in the TPI promoter was determined in 378 unrelated normal subjects from sub Saharan African (n = 103), Caribbean (n = 26), Northern European (n = 57), Mediterranean (n = 55), Middle Eastern (n = 42), Asian Indian (n = 48) and Oriental (n = 47) populations. Five haplotypes were identified: the common haplotype, -5A-8G-24T, -5G, -8A, -5G-8A, and -5G-8A-24G. All, with the exception of the -8A haplotype, were present in geographically dispersed populations. The 5G allele, which was found at varying frequency in all groups, has attained high frequency in the African, Caribbean and Oriental populations. Phylogenetic comparison suggests this may represent the ancestral promoter haplotype. Homozygosity for the -5G-8A haplotype identified in four subjects confirms that these variants are not responsible for a null allele as formerly postulated. Linkage disequilibrium between related TPI promoter haplotypes, -5G, -5G-8A and 5G-8A-24G, and a single nucleotide polymorphism at nt2262 of the TPI gene supports a single ancestral origin for these mutations which precedes the separation of African and non-African populations. PMID- 10453738 TI - Association of the human NPPS gene with ossification of the posterior longitudinal ligament of the spine (OPLL). AB - OPLL (ossification of the posterior longitudinal ligament of the spine) is a common form of human myelopathy with a prevalence of as much as 4% in a variety of ethnic groups. To clarify the genetic factors that predispose to OPLL, we have studied ttw (tiptoe walking), a mouse model that presents ectopic ossification of the spinal ligaments similar to OPLL and have found that the ttw phenotype is caused by the nonsense mutation of the gene encoding nucleotide pyrophosphatase (NPPS), a membrane-bound glycoprotein thought to produce inorganic pyrophosphate, a major inhibitor of calcification and mineralization. To investigate a possible role of NPPS in the etiology of OPLL, we have examined its genetic variations in OPLL patients. A total of 323 OPLL patients was screened by means of polymerase chain reaction/single-strand conformation polymorphism analysis covering all the exons and their surrounding introns, plus about 1.5-kb of the promoter region. We identified ten nucleotide variations in the NPPS gene; five of the alterations caused amino-acid substitutions, and two of them were found specifically in OPLL patients. Subsequently, we performed an association study using these variations and found a significant association of an allele, viz., a deletion of T at a position 11 nucleotides upstream from the splice acceptor site of intron 20 (IVS20-11delT), with OPLL; the proportion of the individuals having this deletion was significantly higher (P = 0.0029) in OPLL patients than in controls, indicating that those who have this variation may be more susceptible to the abnormal ossification of the spinal ligaments. Thus, our study suggests that NPPS plays an important role in the etiology of human OPLL. PMID- 10453739 TI - Molecular cloning and characterization of the human NUDC gene. AB - In both Aspergillus nidulans and the mouse, studies of the nuclear distribution gene C (NudC) have strongly suggested that the NudC protein interacts with NudF, which is the product of NudF, a homologue of human LIS1 (also know as PAFAH1B1), one of the causative genes for classical lissencephaly. We have isolated the human NUDC gene and its two processed pseudogenes. The human NUDC gene is highly conserved and its predicted amino acid sequence shows 94% identity to mouse NudC and 95% identity to rat NudC. The genomic structure of NUDC, its chromosomal localization, and expression pattern in human tissues were characterized. NUDC consists of at least 9 exons ranging from 66 bp to 266 bp in size and 8 introns from 92 bp to 2.0 kb in length, and the total genomic region spans about 8 kb. NUDC was mapped to 1p34-p35 by fluorescence in situ hybridization. Northern analysis showed a major 1.6 kb transcript in all fetal and adult tissues examined. Primers which amplify individual exons of NUDC were developed for mutation analysis. PMID- 10453740 TI - Comparison of complementary and genomic DNA sequencing for the detection of mutations in the HMBS gene in British patients with acute intermittent porphyria: identification of 25 novel mutations. AB - Acute intermittent porphyria (AIP) is a low-penetrant autosomal dominant disorder caused by mutations in the hydroxymethylbilane synthase (HMBS) gene. Direct detection of mutations is becoming the method of choice for the accurate identification of asymptomatic affected individuals within AIP families so that they can be advised to avoid drugs and other compounds that provoke the life threatening acute neurovisceral crises that characterise the condition. We describe a prospective comparison of direct automated sequencing of cDNA (29 patients) or genomic DNA (28 patients) to identify HMBS mutations in 57 patients referred consecutively for mutational analysis; 39 different mutations were identified in 54 patients. The sensitivity of the cDNA and genomic DNA methods was 69% and 95%, respectively, indicating that analysis of genomic DNA provides a higher mutation detection rate. Thirty mutations were restricted to a single family; only one (R173W) occurred in more than three families. Of the mutations (6 missense, 8 splice defects, 10 frameshifts, 1 nonsense), 25 have not been reported previously. One novel mutation (344+33G-->T) was located in a putative intron splice enhancer in intron7. Our results define the extent of allelic heterogeneity and the types (41% missense; 59% truncating) and distribution (35% in exons 10, 12, 14) of HMBS mutations, for AIP in the United Kingdom. PMID- 10453741 TI - Two novel mutations in a cystic fibrosis patient of Chinese origin. AB - Cystic fibrosis is rare in non-Caucasian populations, and in such populations little is known about the spectrum of mutations and polymorphisms in the CFTR gene. We studied a 23-year-old patient of Chinese ethnicity with sweat chloride values of 104 mM/l, pancreatic sufficiency, an FEV1 60% of normal, sputum cultures positive for Staphylococcus aureus and Burkholderia cepacia, and a history of allergic bronchopulmonary aspergillosis. Genetic screening for 31 common CFTR mutations was negative, leading us to search for unknown mutations using single-strand conformation polymorphism and heteroduplex analysis (SSCP/HA). Two novel mutations were detected. In exon 4, a deletion of 8 bp (451458, deltaGCTTCCTA) causes a frameshift and immediately creates a stop codon. In exon 16, mutation 3041G-->A causes the missense change G970D. Functional analysis using an isotopic flux assay indicated that the G970D mutation retains partial function; western blotting indicated that the protein is glycosylated. The patient is heterozygous for the common polymorphisms (2694T/G) in exon 14a and (GATT)6/7 in intron 6a, indicating that these variants arose in ancestors common to Caucasians and Chinese. PMID- 10453742 TI - Spinocerebellar ataxias in Spanish patients: genetic analysis of familial and sporadic cases. The Ataxia Study Group. AB - Autosomal dominant cerebellar ataxias (ADCA) are a clinically heterogeneous group of neurodegenerative disorders caused by unstable CAG repeat expansions encoding polyglutamine tracts. Five spinocerebellar ataxia genes (SCA1, SCA2, SCA3, SCA6 and SCA7) and another related dominant ataxia gene (DRPLA) have been cloned, allowing the genetic classification of these disorders. We present here the molecular analysis of 87 unrelated familial and 60 sporadic Spanish cases of spinocerebellar ataxia. For ADCA cases 15% were SCA2, 15% SCA3, 6% SCA1, 3% SCA7, 1% SCA6 and 1% DRPLA, an extremely rare mutation in Caucasoid populations. About 58% of ADCA cases remained genetically unclassified. All the SCA1 cases belong to the same geographical area and share a common haplotype for the SCA1 mutation. The expanded alleles ranged from 41 to 59 repeats for SCA1, 35 to 46 [corrected] for SCA2, 67 to 77 for SCA3, and 38 to 113 for SCA7. One SCA6 case had 25 repeats and one DRPLA case had 63 repeats. The highest CAG repeat variation in meiotic transmission of expanded alleles was detected in SCA7, this being of +67 units in one paternal transmission and giving rise to a 113 CAG repeat allele in a patient who died at 3 years of age. Meiotic transmissions have also shown a tendency to more frequent paternal transmission of expanded alleles in SCA1 and maternal in SCA7. All SCA1 and SCA2 expanded alleles analyzed consisted of pure CAG repeats, whereas normal alleles were interrupted by 1-2 CAT trinucleotides in SCA1, except for three alleles of 6, 14 and 21 CAG repeats, and by 1-3 CAA trinucleotides in SCA2. No SCA or DRPLA mutations were detected in the 60 sporadic cases of spinocerebellar ataxia, but one late onset patient was identified as a recessive form due to GAA-repeat expansions in the Friedreich's ataxia gene. PMID- 10453743 TI - Molecular analysis of hyperoxaluria type 1 in Italian patients reveals eight new mutations in the alanine: glyoxylate aminotransferase gene. AB - Systematic screening using the SSCP technique followed by sequencing of bands with abnormal mobility derived from the AGXT exons of 15 unrelated Italian patients with primary hyperoxaluria type 1 (PH1) allowed us to characterize both the mutant alleles in each individual. Eight new mutations were identified: C155del, C156ins, G244T, C252T, GAG408ins, G468A, G588A and G1098del. This study demonstrates both the effectiveness of the screening strategy chosen to identify all the mutant alleles and the high degree of allelic heterogeneity in PH1. PMID- 10453744 TI - Intragenic polymorphic missense mutations in the XLRS1 gene in families with juvenile X-linked retinoschisis. PMID- 10453745 TI - Understanding the mechanism of LCAT reaction may help to explain the high predictive value of LDL/HDL cholesterol ratio. AB - Traditionally, lecithin:cholesterol acyltransferase (LCAT) role in the reverse cholesterol transport (RCT) has been considered "antiatherogenic" as the cholesterol esterification is the prerequisite for the formation of mature high density lipoprotein (HDL) particles and may create a gradient necessary for the flow of unesterified cholesterol (UC) from tissues to plasma. However, newer data suggest that a higher esterification rate is not necessarily protective. Here we review the available data on the role of LCAT in RCT and propose that the LCAT mediated esterification of plasma cholesterol promotes RCT only in the presence of sufficient concentrations of HDL2 while this reaction may be atherogenic in the presence of high concentration of plasma low density lipoprotein (LDL) cholesterol Thus, the "protective" or potentially "atherogenic" role of LCAT depends on the quality of HDL and concentration of LDL. This hypothesis is consistent with the known high predictive value of LDL/HDL cholesterol ratio. PMID- 10453746 TI - Participation of coenzyme Q10 in the rejection development of the transplanted heart: a clinical study. AB - Coenzyme Q10 and alpha-tocopherol concentrations were assessed in 28 endomyocardial biopsies from 22 patients and in 61 blood samples from 31 patients after heart transplantation with histologically confirmed signs of rejection. The values were compared to the group of 14 patients with cardiomyopathies of unclear etiology as candidates for heart transplantation. Blood analyses were also compared with 50 healthy persons. Myocardial and blood coenzyme Q10 concentrations were already significantly decreased in the incipient phase of rejection (degree 0-1) and also in rejection phase 1 and 2. In patients without rejection signs myocardial and blood coenzyme Q10 values were similar to those of cardiomyopathic patients. No significant differences were found in alpha tocopherol concentrations in relation to signs of rejection. Increased plasma lipid peroxidation quantified as malondialdehyde production was detected in all groups of transplanted patients. The results contribute to the explanation of some pathobiochemical mechanisms participating in the rejection development of the transplanted heart. PMID- 10453747 TI - Early changes of protein synthesis in myocardium and coronary arteries induced by NO synthase inhibition. AB - The question was addressed whether short-term (4 hour) NO deficiency, inducing an increase in blood pressure in anaesthetized dogs, does influence proteosynthesis in the myocardium and coronary arteries. A potentially positive answer was to be followed by the study of the supporting role of ornithine decarboxylase for the polyamines pathway. N(G)-nitro-L-arginine-methyl ester (L-NAME) (50 mg/kg per hour) was administered i.v. to inhibit NO synthase. After the first L-NAME dose diastolic blood pressure increased from 131.8+/-2.0 to 149.4+/-3.9 mm Hg (p<0.001) and was maintained at about this level till the end of the experiment. Systolic blood pressure only increased after the first dose (from 150.8+/-1.1 to 175.0+/-5.8 mm Hg, p<0.01), returning thereafter to the control level. Similarly, the heart rate declined only after the first dose (from 190.4+/-5.3 to 147.6+/ 4.5 beats/min, p<0.01). Total RNA concentrations increased in the left cardiac ventricle (LV), the left anterior descending coronary artery (LADCA) and left circumflex coronary artery (LCCA) by 15.9+/-0.7, 29.7+/-1.3 and 17.6+/-1.0%, p<0.05, respectively. The same applied to [14C]leucine incorporation (by 86.5+/ 5.0, 33.5+/-2.6, 29.3+/-4.1%, p<0.05, respectively). The above parameters indicated an increase of proteosynthesis in the LV myocardium and both coronary arteries LADCA and LCCA after short-term NO deficiency. Surprisingly, the ornithine decarboxylase activity in the LV myocardium decreased significantly by 40.2+/-1.6% (p<0.01) but the changes were not significant in the coronary arteries. This unexpected finding makes the role of polyamines in increasing proteosynthesis during a pressure overload due to NO deficiency questionable. PMID- 10453748 TI - Heart phospholipid content and fatty acid composition in the rat after feeding different lipid supplemented diets. AB - The heart phospholipid content and fatty acid composition were examined in adult rats after four weeks of feeding lipid-supplemented diets (20 g % w/w) containing sunflower oil-lard (1:1) mixture (SL group) or margarine (M group). Our results showed a decreased cardiolipin content and distribution in both experimental groups and an increased lysophosphatidylcholine and phosphatidylcholine content and distribution in the SL group with a tendency to lower phosphatidylcholine/phospatidylethanolamine ratio in both experimental groups. In the SL group, the content of saturated fatty acids was higher and that of monounsaturated fatty acids was lower than in the control group. The M group showed inverse results. The content of saturated fatty acids was lower and that of monounsaturated was higher than in the control group. Polyunsaturated n-6 fatty acids were decreased in both experimental groups and n-3 fatty acids were increased in the M group. Feeding lipid-supplemented diets reduced n-6/n-3 and 20:4/22:6 ratios in the M group. The polyunsaturated/saturated fatty acid ratio was lower in the SL and higher in indicating the M group than in the control group. Our results are in agreement with the other reports indicating that the heart is sensitive to diet-induced lipid alterations. PMID- 10453749 TI - Long-term follow-up of the tubular secretion of creatinine in renal graft recipients. AB - The differences in glomerular filtration rate (GFR) based on creatinine clearance (Ccr) or obtained by the more exact methods are caused mainly by tubular creatinine secretion. In this study, we monitored creatinine clearance (Ccr), GFR on the basis of polyfructosan renal clearance (C(PF)) and parameters characterizing tubular creatinine secretion (Ccr/C(PF), Ccr - C(PF), Tcr/C(PF) x 100) in 12 individuals with renal grafts (Group A), 12 kidney graft donors for related transplantation (Group B), and in 27 individuals undergoing nephrectomy for a pathological process in one kidney (Group C). In the monitored groups, C(PF) and Ccr values were within the limits consistent with the normal function of a single kidney in a healthy individual. The values characterizing tubular creatinine secretion in Group A did not differ significantly from those obtained in Groups B and C. However, the parameters showed a wide range in all groups. In seven individuals with a renal graft, all the above functional parameters were monitored at three-month intervals for a period of 24 months. Significant differences in the time courses of Ccr and C(PF) due to marked intra-individual fluctuations were found in tubular creatinine secretion. The findings suggest that the rate of tubular creatinine secretion in the renal graft does not differ significantly from that in individuals with a single native (normally functioning) kidney. However, there are large inter-individual differences. The large intra-individual fluctuations in tubular creatinine secretion in the kidney graft result in significant differences in the time courses of Ccr and C(PF) and a possibility of erroneous evaluation of graft function if based exclusively on Ccr. PMID- 10453750 TI - VO2/power output relationship and the slow component of oxygen uptake kinetics during cycling at different pedaling rates: relationship to venous lactate accumulation and blood acid-base balance. AB - In this experiment we studied the effect of different pedalling rates during cycling at a constant power output (PO) 132+/-31 W (mean+/-S.D.), corresponding to 50% VO2 max, on the oxygen uptake and the magnitude of the slow component of VO2 kinetics in humans. The PO corresponded to 50% of VO2 max, established during incremental cycling at a pedalling rate of 70 rev.min(-1). Six healthy men aged 22.2+/-2.0 years with VO2 max 3.89+/-0.92 l.min(-1), performed on separate days constant PO cycling exercise lasting 6 min at pedalling rates 40, 60, 80, 100 and 120 rev.min(-1), in random order. Antecubital blood samples for plasma lactate [La]pl and blood acid-base balance variables were taken at 1 min intervals. Oxygen uptake was determined breath-by-breath. The total net oxygen consumed throughout the 6 min cycling period at pedalling rates of 40, 60, 80, 100 and 120 rev.min(-1) amounted to 7.727+/-1.197, 7.705+/-1.548, 8.679+/-1.262, 9.945+/ 1.435 and 13.720+/-1.862 l, respectively for each pedalling rate. The VO2 during the 6 min of cycling only rose slowly by increasing the pedalling rate in the range of 40-100 rev.min(-1). This increase, was 0.142 l per 20 rev.min(-1) on the average. Plasma lactate concentration during the sixth minute of cycling changed little within this range of pedalling rates: the values were 1.83+/-0.70, 1.80+/ 0.48, 2.33+/-0.88 and 2.52+/-0.33 mmol.l(-1). The values of [La]pl reached in the 6th minute of cycling were not significantly different from the pre-exercise levels. Blood pH was also not affected by the increase of pedalling rate in the range of 40-100 rev.min(-1). However, an increase of pedalling rate from 100 to 120 rev.min(-1) caused a sudden increase in the VO2 amounting to 0.747 l per 20 rev.min(-1), accompanied by a significant increase in [La]pl from 1.21+/-0.26 mmol.l(-1) in pre-exercise conditions to 5.92+/-2.46 mmol.l(-1) reached in the 6th minute of cycling (P<0.01). This was also accompanied by a significant drop of blood pH, from 7.355+/-0.039 in the pre-exercise period to 7.296+/-0.060 in the 6th minute of cycling (P < 0.01). The mechanical efficiency calculated on the basis of the net VO2 reached between the 4th and the 6th minute of cycling amounted to 26.6+/-2.7, 26.4+/-2.0, 23.4+/-3.4, 20.3+/-2.6 and 14.7+/-2.2%, respectively for pedalling rates of 40, 60, 80, 100 and 120 rev.min(-1). No significant increase in the VO2 from the 3rd to the 6th min (representing the magnitude of the slow component of VO2 kinetics) was observed at any of the pedalling rates (-0.022+/-0.056, -0.009+/-0.029, 0.012+/-0.073, 0.030+/-0.081 and 0.122+/-0.176 l.min(-1) for pedalling rates of 40, 60, 80, 100 and 120 rev.min( 1), respectively). Thus a significant increase in [La]pl and a decrease in blood pH do not play a major role in the mechanism(s) responsible for the slow component of VO2 kinetics in humans. PMID- 10453751 TI - Expression of adhesion molecules and effect of disodium cromoglycate treatment in asthmatics. AB - Allergic processes are complex disorders in which inflammatory and immunological mechanisms are involved. Many studies indicate that the adhesion molecules are upregulated in allergic inflammation, and play a critical role in the pathogenesis of allergic inflammation. Modulation of the expression of adhesion molecules may provide a potential new target for therapeutic intervention in allergic diseases. In the present study the changes expression of adhesion molecules CD11a, CD18 (LFA-1), CD54 (ICAM-1) and L-selectin (CD62L) and VLA-4 (CD49d) were analysed by flow cytometry. Serum concentrations of soluble ICAM-1, VCAM-1 and soluble low affinity receptor for IgE concentrations sCD23 were measured by ELISA in atopic patients with mild asthma before and after treatment by disodium cromoglycate (DSCG). The most significant finding was a significant decrease of ICAM-1 expression on monocytes and CD49d on monocytes and lymphocytes as well as an increase of L-selectin expression on monocytes after treatment with DSCG, without any associated effect on CD11a and CD18. The levels of soluble ICAM 1 and VCAM-1 were not changed, only the levels of soluble CD23 that plays a regulatory role in ongoing IgE production, were decreased in asthmatic patients after the treatment with DSCG. These results suggest that DSCG diminishes cell activation. PMID- 10453752 TI - Effect of disodium cromoglycate treatment on peripheral blood mononuclear cell adhesion to cultured endothelium in allergic asthmatics. AB - In this study we have compared the adhesion of peripheral blood mononuclear cells (PBMC) to human umbilical vein endothelial cells (HUVEC) in a healthy control group with two groups of allergic asthmatics, not treated or treated with disodium cromoglycate (DSCG). The adhesion and blocking experiments were performed by the flow cytometric adhesion assay. No differences in the adhesion of lymphocytes were observed in any of the groups. The monocytes obtained from DSCG non-treated patients have shown significant (P < 0.05) enhancement of adhesion to HUVEC in comparison to healthy controls. The treatment of asthmatic patients with DSCG downregulated the monocyte adhesion to cultured endothelial cells (ECs) and this was comparable to the group of normal donors. The DSCG may have a therapeutic effect on the regulation of monocyte adhesion in inflammatory and allergic diseases. The binding ability of untreated asthmatic PBMC to cultured ECs was partially inhibited by monoclonal antibody anti-CD54, suggesting that the increased EC adhesiveness for monocytes from allergic asthmatics may be at least partially dependent on the ICAM-1 adhesion pathways. Our results also indicate that the blocking agent anti-CD18 was not essential for monocyte endothelial interactions in allergic asthma. PMID- 10453753 TI - Granulocyte and plasma cytokine activity in acute cadmium intoxication in rats. AB - Changes in the number and ex vivo function of peripheral blood neutrophils were investigated following intraperitoneal administration of cadmium-chloride in rats. Besides a dose-dependent increase in the number of peripheral blood neutrophils, changes were found in the functional state of isolated polymorphonuclear leukocytes (PMNs). Increased spontaneous adhesion and activation, and TNF activity in a conditioned medium were observed in cultures of granulocytes in comparison to granulocytes from control (saline-treated) animals. Increased levels of plasma activity of inflammatory cytokines, tumor necrosis factor (TNF) and interleukin-6 (IL-6) were noted following cadmium administration. Cytological signs of pulmonary inflammation were revealed histologically and the majority of neutrophils recovered from the lungs by enzyme digestion exhibited a capacity of nitroblue tetrazolium (NBT) reduction. Our data demonstrate that acute cadmium intoxication leads to a systemic inflammatory response characterized by numerical and functional changes in the granulocyte compartment and to increased levels of inflammation-related cytokine activity in the circulation. Correlations between the increased number of peripheral blood neutrophils and IL-6 plasma activity (r=0.776, p<0.00001) and the number of neutrophils recovered from the lung tissue (r=0.893, p<0.00001) suggested that systemic cadmium-induced inflammation might be involved in the pulmonary toxicity of cadmium. PMID- 10453754 TI - Time course of isolated rat fundus response to muscarinic agonists: a measure of intrinsic efficacy. AB - The establishment of a dose-response relationship and its quantification is the usual procedure for analysing drug action on an isolated organ. However, the time course of the effect seems to be an inherent characteristic of the agonist which produces it. In our study, we have analyzed the time-response curves of four cholinergic agonists (acetylcholine, methacholine, carbachol and bethanechol) which produce tonic contractions of the isolated rat gastric fundus. The order of affinity of agonists to muscarinic receptors on the rat fundus were carbachol > bethanechol > methacholine > acetylcholine (K(A) values: 46 +/- 12, 84 +/- 21, 380 +/- 110 and 730 +/- 120 nM, respectively). The effective concentrations which produced 60% of the maximal response (EC60) were used for establishing the time response curves. The time-response curves were also recorded after partial alkylation of muscarinic receptors with phenoxybenzamine, after exposure of the isolated rat fundus to physostigmine and after addition of supramaximal concentrations of the agonists. The experimental time-response curve for acetylcholine was on the extreme left, followed by curves for methacholine, bethanechol and carbachol, respectively. Phenoxybenzamine and supramaximal doses of the agonists did not change the order of response development in time, but supramaximal doses shifted all curves to the left and phenoxybenzamine shifted all time-response curves to the right. Only physostigmine shifted the time response curve for methacholine to the right. The results of our study suggest that the response rate of the isolated rat gastric fundus to cholinergic agonists depends on the intrinsic activity of these agents, but not on their affinity for muscarinic receptors. PMID- 10453755 TI - The influence of superovulation preparations on the levels of catecholamines in eminentia mediana, the pituitary and pineal gland of the sheep. AB - The influence of hormonal superovulation preparations of FSH (450 IU) or PMSG (1500 IU), on the levels of catecholamines (dopamine, norepinephrine and epinephrine) was studied in the oestrus period using radioenzymatic methods. The administration of FSH caused a significant increase in the concentrations of norepinephrine (NE) and epinephrine (EPI) in eminentia mediana (EM) of sheep (p<0.001 and p<0.01, respectively). The pituitary gland exhibited an increase in the level of norepinephrine after administration PMSG while no marked changes were recorded for epinephrine and dopamine (DA). The administration of FSH affected the increase in pituitary epinephrine (p<0.01). The hormonal stimulation by FSH resulted in a marked decrease of dopamine (p<0.05) as well as in a significant increase of norepinephrine (p<0.05) and epinephrine (p<0.05) in the epiphysis. The comparison of the effect of hormonal preparations on the changes in catecholamine levels showed that the effect of FSH was observed mostly in eminentia mediana and the pituitary gland while that of PMSG was recorded in the epiphysis. PMID- 10453756 TI - Brain cell RNA: sexual differences in the rat. AB - Sexual differences in the total content of ribosomal RNA, established by cytophotometric measurements in neurones from selected brain regions, were studied in rats of the Wistar strain. In females of reproductive age, cyclic changes of RNA were synchronized with their oestrous cycle, the values being higher in the oestrous phase than in dioestrus. These changes were observed in pyramidal cells of the hippocampus and of the frontal cortex, in cells of anterior thalamus, of ventromedial and lateral hypothalamus and of tuberculi olfactorii. However, cycling cells were not disclosed in septum and thalamus posterior. A dependence upon the actual level of ovarial hormones was found in ventromedial hypothalamic cells only. In general, the RNA values in males of the same age corresponded to values of dioestrous females. The differences between newborn and 7-day-old pups were not marked enough and did not allow to define the critical period responsible for initiation of this sexual difference. In senescent rats, this difference persisted. During the stable phase of long lasting dioestrus, the total RNA content in cells of the frontal cortex, hippocampus and hypothalamus was higher in females than in males of the same age which may suggest a faster reduction of this substance in aged males. The prolonged influence of oestrogens in the oestrous phase of the climacteric period (preceding the permanent dioestrus) decreased the RNA values in hippocampal and hypothalamic neurones even below the level established during the permanent dioestrus (and thus reached male values). On the contrary, in frontal cortical neurones, the female values remained higher in the permanent dioestrus as well as during long-lasting oestrus. A discussion concerns the possible participation of genetic determination and of the actual state of ovarial hormones in the manifestation of sexual differences in brain cells of the rat. PMID- 10453757 TI - Reciprocal connections between the red nucleus and the trigeminal nuclei: a retrograde and anterograde tracing study. AB - An anterograde biocytin and a retrograde WGA-colloidal gold study in the rat can provide information about reciprocal communication pathways between the red nucleus and the trigeminal sensory complex. No terminals were found within the trigeminal motor nucleus, in contrast with the facial motor nucleus. A dense terminal field was observed in the parvicellular reticular formation ventrally to the trigeminal motor nucleus. The parvicellular area may be important for the control of jaw movements by rubrotrigeminal inputs. On the other hand, the contralateral rostral parvicellular part of the red nucleus receives terminals from the same zone in the rostral part of the trigeminal sensory complex, where retrogradely labelled neurones were found after tracer injections into the red nucleus. Such relationships could be part of a control loop for somatosensory information from the orofacial area. PMID- 10453758 TI - The influence of angiotensin II on sex-dependent proliferation of aortic VSMC isolated from SHR. AB - The growth response to angiotensin II (Ang II) was studied using cultured vascular smooth muscle cells (VSMC) isolated from the aortae of male and female spontaneously hypertensive rats (SHR). Systolic and mean arterial blood pressure of 10-week-old males was significantly higher when compared to age-matched females. The specific growth rate of male VSMC was significantly higher on the third and sixth day after synchronisation. Angiotensin II in concentration 10(-7) M stimulated the specific growth rate only in male VSMC during the exponential phase of growth. Moreover, doubling time was 3 hours shorter in male VSMC in comparison with the females. Our results suggest that both the increased specific growth rate and augmented growth-response of male VSMC to Ang II may explain the higher sensitivity of males to hypertensive stimuli. PMID- 10453759 TI - Cytosolic free calcium response to angiotensin II in aortic VSMC isolated from male and female SHR. AB - Previous in vitro studies have shown that vascular smooth muscle cells (VSMC) isolated from the aortae of male spontaneously hypertensive rats (SHR) proliferate more rapidly than those obtained from female SHR. Sex-dependent differences of cytosolic free calcium concentration ([Ca2+]i) were therefore studied in VSMC under basal conditions and after the stimulation by different concentrations of angiotensin II (Ang II). No significant difference in basal [Ca2+]i was found in VSMC from male and female SHR. Angiotensin II significantly increased [Ca2+]i in VSMC from both genders. This [Ca2+]i rise elicited by 10(-7) and 10(-9) M Ang II was more pronounced in cells isolated from males than in those from females. This difference may be attributed to greater mobilisation of intracellular calcium stores in male VSMC. It can be concluded that the cytosolic free calcium response to angiotensin II is augmented in VSMC of male SHR, which also grow more rapidly in response to this peptide hormone. PMID- 10453760 TI - Impaired adrenal reserve in men with spinal cord injury: results of low- and high dose adrenocorticotropin stimulation tests. AB - OBJECTIVE: To use low- and high-dose adrenocorticotropin (ACTH) tests to assess adrenal reserve in men with spinal cord injury (SCI). DESIGN: After an overnight fast, 1 microg and 200 microg ACTH were injected intravenously at time 0 and 60 minutes between 8 AM and 9 AM. Blood was withdrawn at 30-minute intervals from time 0 to 120 minutes. SETTING: All participants were recruited from the outpatient clinic of a university hospital that is a tertiary referral center. PARTICIPANTS: Forty-two men with traumatic neurologically complete SCI that had occurred more than 1 year before the study. MAIN OUTCOME MEASURES: Serum cortisol response to ACTH at times 0, 30, 60, 90, and 120 minutes. RESULTS: Twenty subjects had a serum cortisol response of <20 microg/dL 30 minutes after a 1 microg ACTH injection; 10 of these remained at this level at 30 minutes after a 200-microg ACTH injection. CONCLUSION: There is a high prevalence of impaired adrenal reserve in persons with chronic SCI. The 1-microg (low dose) ACTH test is more sensitive for detecting subclinical adrenal insufficiency than is the 200 microg (high dose) ACTH test. PMID- 10453761 TI - Correlates of life satisfaction among persons with spinal cord injury. AB - OBJECTIVE: To analyze the correlates of life satisfaction for individuals with spinal cord injury (SCI). STUDY DESIGN: Survey; follow-up of subjects studied prospectively since onset of injury. PARTICIPANTS: A total of 2,183 persons with SCI, from 1 to 20 years postinjury, self-selected for annual research (and clinical) follow-up by one of 18 model systems of SCI care. RESULTS: Life satisfaction, as measured with the Satisfaction With Life Scale (SWLS), is associated with several demographic, social, functional, and clinical characteristics. Stepwise linear regression analysis resulted in a predictor model that included the following: sex (beta weight: .07; p < .001); number of rehospitalizations in the last year (-.05; p < .05); years since injury (.13; p < .0001); sociocognitive disability as measured with the Functional Independence Measure (.06; p < .01); and three handicap components, as measured with the modified Craig Handicap Assessment and Reporting Technique: mobility (.26; p < .0001); occupation (.10; p < .001); and social integration (.11; p < .0001). Impairment (level of injury) contributed indirectly, through its impact on motor disability. Racial/ethnic group membership, motor disability, and education contributed indirectly, through their effects on handicap. CONCLUSIONS: Life satisfaction after SCI can be reliably measured by means of the SWLS. Correlates of subjective well-being parallel those suggested by earlier studies and those for the population at large. The effects of life satisfaction on social participation, health, and other aspects of life need further study. PMID- 10453762 TI - Performance of health-related quality-of-life instruments in a spinal cord injured population. AB - OBJECTIVE: General health-related quality-of-life (HRQoL) surveys have not been well tested in populations with spinal cord injury (SCI). This study evaluated the performance of 5 such instruments. DESIGN: A cross-sectional survey with instruments administered in random order during computer-assisted interviews. SETTING: A midwestern US veteran SCI program. SUBJECTS: One hundred eighty-three veterans with SCI ranging in age from 21 to 81 yrs (mean = 50.5). MEASURES: The Behavioral Risk Factor Surveillance System (BRFSS) HRQoL modules, the Quality of Well-Being scale (QWB), the Medical Outcomes Study Short-Form 36 and Short-Form 12 (SF-36, SF-12), and instrumental activities of daily living (IADL). RESULTS: Construct validity was supported by scores from the QWB, IADL, and physical health measures of the BRFSS and SF-36 showing greater impairment for quadriplegia than paraplegia. Similar constructs on the SF-36 and BRFSS were more strongly correlated than between the IADL and QWB; eg, correlation between the SF 36 Vitality scale and the BRFSS "Days full of energy" question was r = .789 (p < .01), whereas correlation between the IADL and QWB was r = -.454 (p < .01). Longer surveys (SF-36, QWB) were rated lower in subject acceptability. CONCLUSIONS: These instruments have potential for research use among patients with SCI. More studies are needed to explore the best use of instruments with apparently different domains. PMID- 10453763 TI - Return to school after spinal cord injury. AB - OBJECTIVE: To determine the typical time elapsed between discharge from an inpatient spinal cord injury (SCI) rehabilitation program and the physical return to school, and to identify barriers faced by patients attempting to return to school. STUDY DESIGN: A retrospective review of all patients ages 18 years or younger who sustained SCI between 1989 and 1995, with resultant paraplegia or tetraplegia, and who attended either primary or secondary school, completed their inpatient rehabilitation at our regional SCI center, and were using a wheelchair when discharged from the hospital. RESULTS: Fifteen of 16 eligible patients agreed to participate. The median time for subjects with paraplegia to return to school after hospital discharge was 10 days; subjects with tetraplegia required a median of 62 days. Architectural and transportation barriers that patients encountered were identified. CONCLUSION: Individuals with SCI return to school relatively soon after discharge from the hospital. Barriers do not prohibit a return to school, but they are problematic. PMID- 10453764 TI - Functional outcome in children with multiple trauma without significant head injury. AB - OBJECTIVE: To assess functional outcome and describe disability at discharge in children who have had trauma without significant head injury. DESIGN: Retrospective cohort. SETTING: National Pediatric Trauma Registry, 1988-1994. PARTICIPANTS: Patients of ages 7 to 18 years with Glasgow Coma Scale (GCS) 13 to 15 without significant anatomic head inJury. RESULTS: Functional Independence Measure (FIM) at discharge was used to assess patient outcome. There were 13,649 children meeting study criteria who had sustained 34,254 injuries. Fractures constituted 30% of all injuries. As measured by FIM, 1,522 (11.2%) patients had mild disability at discharge; 1,983 (14.5%) had moderate disability. After adjustment for age and injury severity, children with lower extremity fractures were more likely to be discharged with functional limitations than those without (relative risk, 5.43; 95% confidence interval: 5.06, 5.84). Of children with moderate disability at discharge, less than 50% were referred for rehabilitation evaluation and less than 25% for physical therapy. CONCLUSION: Functional dependence is present in a large proportion of injured children, even without significant head injury. Rehabilitation and other services may be underused in this population. Further study is required to fully assess the degree and duration of disability in these patients. PMID- 10453765 TI - Osteogenesis imperfecta in childhood: impairment and disability--a follow-up study. AB - OBJECTIVES: To evaluate differences over time (mean follow-up, 14 months) on impairment parameters (range of joint motion and muscle strength), functional limitation parameters (functional ability), and disability parameters (caregiver assistance in achieving functional skills) in osteogenesis imperfecta (OI), related to the different types of the disease. DESIGN: A prospective, descriptive study. MATERIALS AND METHODS: Fifty-four children with OI and their parents participated at the start of the study. At the end, 44 children participated in the assessment of functional skills and 42 of them participated in clinical assessment (OI type I, n = 19; OI type III, n = 13; OI type IV, n = 10). Range of joint motion was measured by means of goniometry. Generalized hypermobility was scored according to Bulbena. Manual muscle strength was scored by means of the MRC grading system. The level of ambulation was scored according to Bleck, and functional skills and caregiver assistance were scored with the Pediatric Evaluation of Disability Inventory. RESULTS: The different types of OI have impact on impairment, functional limitation, and disability. Almost all impairment parameters did not change significantly over time, whereas some disability parameters seemed to improve significantly. CONCLUSIONS: Impairment parameters in OI are presumably not always preconditions for functional limitation and disability. A 1-year follow-up revealed no significant changes in impairment parameters, whereas some disability parameters improved. Treatment strategies in OI should, therefore, focus primarily on improving functional ability, with respect to the natural course of the disease, and not only on impairment parameters. PMID- 10453766 TI - Electrodiagnosis in spinal cord injured persons with new weakness or sensory loss: central and peripheral etiologies. AB - OBJECTIVE: To assess the prevalence and causes of late neurologic decline of persons with spinal cord injury (SCI). DESIGN: Retrospective review of persons with SCI over a 9-year period. Those with complaints of new weakness or sensory loss were grouped into three categories based on clinical examination, electrodiagnosis, and imaging: (1) central pathology (ie, brain, spinal cord, or nerve root); (2) peripheral pathology (plexus or peripheral nerve); or (3) no identifiable etiology. The specific diagnoses of late neurologic decline were identified. SETTING: Regional Veterans Affairs Spinal Cord Injury Service. PATIENTS: Five hundred two inpatient and outpatient adults with SCI. RESULTS: Nineteen percent of the study population complained of new weakness and/or sensory loss. Neurologic abnormalities were noted in 13.5%, 7.2% with central and 6.4% with peripheral causes. The most common pathologies were posttraumatic syringomyelia (2.4%) and cervical (1.6%) and lumbosacral (1.2%) myelopathy/radiculopathy. A specific etiology was not determined in 6 cases (1.6%). Peripheral involvement was mostly from ulnar nerve entrapment (3.4%) and carpal tunnel syndrome (3.0%). CONCLUSIONS: Late-onset neurologic decline is common after SCI and can result from central or peripheral pathology. Regular neurologic monitoring of SCI patients is recommended, since many with neurologic decline respond favorably if diagnosed and treated early. PMID- 10453767 TI - Wheelchair pushrim kinetics: body weight and median nerve function. AB - OBJECTIVES: Individuals who use manual wheelchairs are at high risk for median nerve injury and subsequent carpal tunnel syndrome (CTS). To gain a better understanding of the mechanism behind CTS in manual wheelchair users, this study examined the relation between (1) pushrim biomechanics and function of the median nerve, (2) pushrim biomechanics and subject characteristics, and (3) median nerve function and subject characteristics. DESIGN: Case series. SETTING: Biomechanics laboratory and an electromyography laboratory. PARTICIPANTS: Thirty-four randomly recruited individuals with paraplegia who use a manual wheelchair for mobility. INTERVENTION: Subjects propelled their own wheelchair on a dynamometer at 0.9m/sec and 1.8m/sec. Bilateral biomechanical data were obtained using a force- and moment-sensing pushrim and a motion analysis system. Bilateral nerve conduction studies focusing on the median nerve were also completed. MAIN OUTCOME MEASURES: Pearson's correlation coefficients between subject characteristics, median nerve conduction studies, and propulsion biomechanics; a regression model of nerve conduction studies incorporating subject characteristics and pushrim biomechanics. RESULTS: Subject weight was significantly related to median nerve latency (r = .36, p = .03) and median sensory amplitude (r = -.43, p = .01). Height was also significantly related to median sensory amplitude (r = -.58, p = .01). Subject weight was significantly related to the peak resultant force applied to the pushrim (r = .59, p < .001). Height, weight, and weight-normalized pushrim forces were successfully incorporated into a linear regression model predicting median sensory amplitude (r = .63, p < .05) and mean median latency (r = .54, p < .05). CONCLUSION: This study found subject weight to be related to pushrim forces and median nerve function. Independent of subject weight, pushrim biomechanics were also related to median nerve function. Through weight loss and changes in pushrim biomechanics, it may be possible to prevent median nerve injury in manual wheelchair users. PMID- 10453768 TI - Home-based multicomponent rehabilitation program for older persons after hip fracture: a randomized trial. AB - OBJECTIVE: To determine whether a home-based systematic multicomponent rehabilitation strategy leads to improved outcomes relative to usual care. DESIGN: A randomized controlled trial with 12 months of follow-up. SETTING: General community; 27 home care agencies. PARTICIPANTS: Three hundred four nondemented persons at least 65 years of age who underwent surgical repair of a hip fracture at two hospitals in New Haven, CT, and returned home within 100 days. INTERVENTION: Systematic multicomponent rehabilitation strategy addressing both modifiable physical impairments (physical therapy) and activities of daily living (ADL) disabilities (functional therapy) versus usual care. MAIN OUTCOME MEASURES: A battery of self-report and performance-based measures of physical and social function. RESULTS: There was no significant difference in the proportion of participants in the two groups who recovered to prefracture levels in self care ADL at 6 months (71% vs 75%) or 12 months (74% in both groups) or in home management ADL at 6 months (35% vs 44%) or 12 months (44% vs 48%). There also was no difference between the two groups in social activity levels, two timed mobility tasks, balance, or lower extremity strength at either 6 or 12 months. Compared with participants who received usual care, those in the multicomponent rehabilitation program showed slightly greater upper extremity strength at 6 months (p = .04) and a marginally better gait performance (p = .08). CONCLUSIONS: The systematic multicomponent rehabilitation program was no more effective in promoting recovery than usual home-based rehabilitation. Compared with previous cohorts, however, participants randomized to usual care in our study received more rehabilitative and home care services and experienced a higher rate of recovery. This finding is important given the current pressures to reduce home services. The challenge is to determine the composition and duration of rehabilitation and home services that will ensure optimal functional recovery most efficiently in older persons after hip fracture. PMID- 10453769 TI - Stretching and strengthening exercises: their effect on three-dimensional scapular kinematics. AB - OBJECTIVE: To quantitatively evaluate the effects of commonly used shoulder exercises on shoulder kinematics and resting posture. STUDY DESIGN: A repeated measures design was used with measurements performed before and after a 6-week exercise program. METHOD: Twenty asymptomatic subjects with forward shoulder posture were recruited. Stretching exercises for the pectoral muscles and resisted strengthening exercises for the scapular retractors and elevators and the glenohumeral abductors and external rotators were performed three times per week for 6 weeks. A three-dimensional electromechanical digitizer was used to measure thoracic inclination and scapular orientation and position. These measurements were taken with the arm (1) at the side, (2) abducted to 90 degrees, and (3) at maximal abduction. The isometric force of glenohumeral external and internal rotation and horizontal abduction and adduction were measured with a hand-held dynamometer. All subjects were tested before and after the 6-week exercise program. Hotelling's T2 and paired t tests were used for data analysis. RESULTS: The strength of horizontal abduction and internal and external rotation increased after exercise (p < .01). The anterior inclination of the thoracic spine decreased, and the glenohumeral contribution to arm elevation increased (p < .01). Resting scapular posture did not change. As the arm was abducted to 90 degrees, the scapula showed less upward rotation and less superior translation after the exercise program (p < .01). CONCLUSION: The exercise program improved muscle strength, produced a more erect upper trunk posture, increased scapular stability, and altered scapulohumeral rhythm. PMID- 10453770 TI - Rectus femoris: its role in normal gait. AB - OBJECTIVE: To analyze the role of the rectus femoris muscle in nondisabled gait at various walking velocities using fine-wire dynamic electromyography. DESIGN: Descriptive study. Fine-wire electromyography data were collected from the rectus femoris during level walking at four walking speeds. Rectus femoris activity patterns in the loading response phase and the pre- and initial-swing phase of the gait cycle were compared using paired t tests. SETTING: A gait laboratory. SUBJECTS: Ten nondisabled adult volunteers. MAIN OUTCOME MEASURES: Amplitude of rectus femoris activity in the loading response phase and the pre- and initial swing phase during walking at four speeds. RESULTS: There was a bimodal pattern of rectus femoris activity in all subjects, at all speeds, in both phases, with high variability in the onsets, durations, and amplitudes of activity, and paired t tests revealed no significant differences (p > .05) between phases at any walking speed. CONCLUSION: Activity in the rectus femoris in the pre- and initial swing phase in nondisabled individuals at all speeds suggests that similar activity detected in individuals with stiff-legged gait may not be inappropriate. PMID- 10453771 TI - Healing times of pedal ulcers in diabetic immunosuppressed patients after transplantation. AB - OBJECTIVE: To compare the healing time of neuropathic plantar ulcers treated by total-contact casting (TCC) in diabetic, immunosuppressed patients after organ transplantation with the healing time of plantar ulcers in control nonimmunosuppressed patients. DESIGN: A case-control design with the control group matched for age, race, sex, body dimensions (height, weight, and body mass index), presence of sensory neuropathy, foot deformity presence and location, and pedal ulcer area and depth. SETTING: An outpatient physical therapy clinic in a regional tertiary-care hospital and academic medical center. PARTICIPANTS: Nine patients with chronic diabetes mellitus and a previous organ transplantation who were currently receiving lifelong immunosuppressive drug therapy were treated for a neuropathic plantar ulcer by means of TCC. Fourteen group-matched control subjects with diabetes mellitus and a plantar ulcer but who had never had an organ transplantation and were not taking immunosuppressive agents were also studied. INTERVENTIONS: TCC with partial weight-bearing using an assistive device until ulcers healed. MAIN OUTCOME MEASURE: Healing time was defined as the number of days in the total-contact cast until the skin completely closed. RESULTS: All diabetic foot ulcers healed with casting. Immunosuppressed/transplanted patients healed in a mean time of 111 +/- 25 days; ulcers of control subjects healed in 47 +/- 18 days (p < .05). All patients returned to ambulation using prescribed therapeutic footwear. None of the patients required a lower extremity amputation throughout the follow-up period. CONCLUSIONS: TCC is a highly effective and rapid method of healing neuropathic pedal ulcers in diabetic immunosuppressed/transplantation patients, although it may take several weeks longer than it would for patients who were not immunocompromised. PMID- 10453772 TI - Endurance times for low back stabilization exercises: clinical targets for testing and training from a normal database. AB - OBJECTIVE: To establish isometric endurance holding times, as well as ratios between torso extensors, flexors, and lateral flexors (stabilizers), for clinical assessment and rehabilitation targets. DESIGN: Simple measurement of endurance times in four tests performed in random order by a healthy cohort. To measure reliability, a subsample also performed the tests again 8 weeks later. SETTING: University laboratory. PARTICIPANTS: Seventy-five young healthy subjects (31 men, 44 women). RESULTS: Women had longer endurance times than men for torso extension, but not for torso flexion or for the "side bridge" exercise, which challenges the lateral flexors (stabilizers). Men could sustain the "side bridge" for 65% of their extensor time and 99% of their flexion time, whereas women could sustain the "side bridge" for only 39% of their extensor time and 79% of their flexion time. The tests proved reliable, with reliability coefficients of >.97 for the repeated tests on 5 consecutive days and again 8 weeks later. CONCLUSION: Healthy young men and women possess different endurance profiles for the spine stabilizing musculature. Given the growing support for quantification of endurance, these data of endurance times and their ratios between extensor, flexor, and lateral flexor groups in healthy normal subjects are useful for patient evaluation and for providing clinical training targets. PMID- 10453773 TI - Thoracic position effect on shoulder range of motion, strength, and three dimensional scapular kinematics. AB - OBJECTIVES: To determine the effect of thoracic posture on scapular movement patterns, active range of motion (ROM) in scapular plane abduction, and isometric scapular plane abduction muscle force. STUDY DESIGN AND METHOD: Repeated measures design. There were 34 healthy subjects (mean age, 30.2 yrs). Each subject was positioned and stabilized while sitting in both erect and slouched trunk postures. In each sitting posture a three-dimensional electromechanical digitizer was used to measure thoracic flexion and scapular position and orientation in three planes. Measurements were taken with the arm (1) at the side, (2) abducted to horizontal in the scapular plane, and (3) at maximum scapular plane abduction. In each posture, isometric abduction muscle force was measured with the arm at the side and abducted to horizontal in the scapular plane. RESULTS: In the slouched posture, the scapula was significantly more elevated in the interval between 0 to 90 degrees abduction. In the interval between 90 degrees and maximum abduction, the slouched posture resulted in significantly less scapular posterior tilting. There was significantly less active shoulder abduction ROM in the slouched posture (mean difference = 23.6 degrees +/- 10.7 degrees). Muscle force was not different between slouched and erect postures with the arm at the side, but with the arm horizontal muscle force was decreased 16.2% in the slouched position. CONCLUSION: Thoracic spine position significantly affects scapular kinematics during scapular plane abduction, and the slouched posture is associated with decreased muscle force. PMID- 10453774 TI - Evaluation and rehabilitation of patients with adult motor neuron disease. AB - Adult motor neuron disease (amyotrophic lateral sclerosis [ALS]) is a neurodegenerative disorder characterized by loss of motor neurons in the cortex, brain stem, and spinal cord, manifested by upper and lower motor neuron signs and symptoms affecting bulbar, limb, and respiratory musculature. Clinically, the disease course is characterized by progressive weakness, atrophy, spasticity, dysarthria, dysphagia, and respiratory compromise, ultimately resulting in death or mechanical ventilation in the vast majority of patients. Patterns of presentation and pathological features of the disease, along with clinical and electrophysiologic criteria for diagnosis, are discussed in this review. Since 8% to 22% of patients survive more than 10 years without ventilator use, meticulous medical and rehabilitation management is extremely important to ensure optimal health and quality of life in these patients. Major issues in the care of individuals with ALS include weakness and spasticity, impairments in activities of daily living and mobility, communication deficits and dysphagia in those with bulbar involvement, respiratory compromise, fatigue and sleep disorders, pain, and psychosocial distress. Research in ALS changes rapidly, but is currently focused on potential etiologic factors such as glutamate excitotoxicity, role of oxidative stress, autoimmunity to calcium channels, and cytoskeletal abnormalities, as well as related treatment initiatives including glutamate modulators, neurotrophic factors, antioxidants, antiapoptotic factors, and gene therapy. Recently, mutations in the gene encoding Cu/Zn superoxide dismutase were identified in a subset of familial ALS patients. Riluzole, a glutamate antagonist and Na-channel blocker, became the only drug currently approved for treatment of ALS after studies showed a small positive effect on survival. Until a definitive treatment or cure for ALS is found, the multifaceted rehabilitation team approach remains the best hope for improving health and survival in this devastating illness. PMID- 10453775 TI - Electrophysiologic recovery after vitamin E-deficient neuropathy. AB - A case report is presented of an electrophysiologic recovery from vitamin E deficient neuropathy after treatment with water-soluble vitamin E in a patient with chronic hepatobiliary disease. The patient was a 64-year-old man who had experienced progressive difficulty in ambulation, with ataxia, over the previous 3 years. The symptoms were associated with pain, tingling sensation in the extremities, and reduced fine motor activity. The patient had chronic hepatobiliary disease, with recurrent cholangitis and external drainage of bile acid through a T-tube for more than 20 years. Vitamin E level was barely detectable (<0.5 mg/L). Sensory conduction was absent in both sural nerves. Other sensory and motor conduction studies in the upper and lower extremities showed decreased amplitude. The patient was treated with water-soluble vitamin E. After 4 months of therapy, his ambulation function improved, but pain and tingling sensation in both hands remained. Sensory nerve action potentials appeared in both sural nerves, and amplitudes of other sensory nerves were increased. In a second follow-up study after 9 months, all of the evaluated parameters in the nerve conduction studies, as well as the vitamin E level, were normal. The authors conclude that vitamin E-deficient neuropathy is reversible and electrophysiologic recovery can occur with water-soluble vitamin E therapy. PMID- 10453776 TI - Analysis of the forces and position required for direct laryngoscopic exposure of the anterior vocal folds. AB - The sniffing position is traditionally considered optimal for direct laryngoscopic examination of the vocal folds. This study examined head and neck positions associated with ideal exposure of the anterior glottal commissure with a variety of laryngoscopes. A prospective investigation was done in 20 patients by comparing the force required to expose the anterior vocal folds by utilizing 3 head and neck positions with 3 different-sized tubular laryngoscopes. The completeness of anterior glottal exposure was rated and the force required to achieve this exposure was measured with a strain gauge. Three positions relating the atlanto-occipital and cervicothoracic vertebrae were analyzed: 1) extension extension. 2) sniffing: extension-flexion, and 3) flexion-flexion. Head and neck position and laryngoscope size were both statistically significant factors for achieving complete anterior vocal fold exposure. Regardless of the laryngoscope, the number of patients in whom complete exposure could be achieved increased gradually when the position was changed from extension-extension to extension flexion to flexion-flexion. Complete exposure was inversely related to larger laryngoscope size. According to the data herein, the flexion-flexion position provides the best glottal exposure for endotracheal intubation in those patients who are anatomically predisposed to difficulty in direct examination of the glottis. Because this places the laryngoscope lumen in a vertical position, this position is inappropriate for microlaryngoscopy. The study reinforced the concept that the sniffing position is the optimal position for microlaryngoscopy because it enables the use of the largest-lumened laryngoscope. This facilitates ideal exposure of the anterior vocal folds, which is necessary for phonomicrosurgery. PMID- 10453777 TI - Laryngopharyngeal sensory discrimination testing and the laryngeal adductor reflex. AB - Laryngopharyngeal sensory capacity has been determined by endoscopically administering air pulse stimuli to the mucosa innervated by the superior laryngeal nerve and asking the patient if he or she feels the stimulus. A potential shortcoming of this psychophysical testing (PT) procedure is that it is a subjective test, and patients with impaired cognition may not be able to perform the required task. In the search for an objective measure of laryngeal sensory function, we have observed that the laryngeal adductor reflex (LAR) is evoked at stimulus intensities similar to those capable of eliciting the psychophysical, or perceptual, response. The purpose of this study is to determine if the threshold for eliciting the LAR is the same as that of the sensory threshold. A specially designed endoscope was used to present air pulse stimuli (range 0.0 to 10 mm Hg) to the laryngopharynx in 20 healthy subjects and in 80 patients with dysphagia, using both PT and the LAR. The patients had a variety of underlying diagnoses, with stroke and chronic neurologic disease predominating (n = 65). In the control group and in the group of patients with dysphagia, there was no statistically significant difference between the median laryngopharyngeal sensory thresholds whether we used PT or the LAR (p>.05, Wilcoxon signed-rank test). The intraclass correlation for the total sample was .999 (U = .999, L = .998). Since psychophysical and sensorimotor reflex thresholds were not statistically significantly different and the intraclass correlation was close to a perfect correlation, we conclude that the LAR can be used as an objective and accurate clinical method of endoscopically assessing laryngopharyngeal sensory capacity. PMID- 10453778 TI - Severe pharyngeal stenosis treated with inferiorly based sternocleidomastoid myocutaneous flap. AB - Severe pharyngeal stenosis is a debilitating condition associated with apnea and dysphagia. Treatment options include local flaps and free mucosal grafts. We present 2 cases of severe stenosis. apparently from adenotonsillectomy, that failed more conservative repairs. Both were treated successfully with a sternocleidomastoid myocutaneous flap rotated in through a lateral pharyngotomy. No further treatment has been required. Technical considerations and operative planning are discussed. PMID- 10453779 TI - Prevention of airway complications in thyroplasty patients requiring endotracheal intubation. AB - Patients who have undergone silicone vocal cord medialization and require additional surgery are at risk for airway complications. There is a narrowed glottic aperture that may be prone to develop postoperative laryngeal edema and prosthesis extrusion. This study was designed to assess the management of this difficult airway and to determine the frequency of postintubation complications. We identified 82 patients who had undergone vocal cord medialization with silicone implants between 1991 and 1995. Seventeen of these patients underwent additional surgical procedures requiring general anesthesia. A retrospective review of these patients' charts was performed to determine the management of the airway and the incidence of postintubation complications. There were no postintubation complications in the 17 patients who were studied. The duration of surgery ranged from 40 minutes to 4 hours 15 minutes. Two patients were ventilated via bronchoscope, and 15 patients were intubated orally. The endotracheal tubes ranged from size 6 to size 9 (median size 8). None of the patients required perioperative steroids. All patients were successfully extubated in the recovery room. No patients required intubation or tracheotomy, and there were no implant extrusions. In this study, the incidence of postintubation airway complications in patients who had undergone previous thyroplasty was minimal. Nevertheless, the potential for airway compromise exists. We recommend preoperative discussion with the anesthesiologist, atraumatic intubation with a small endotracheal tube, and diligent observation for airway compromise. PMID- 10453780 TI - Fat implantation into Reinke's space: a histologic and stroboscopic study in the canine. AB - The scarred vocal fold is a cause of persistent dysphonia after laryngeal trauma, microsurgery, and tumor resection. The loss of Reinke's space with scar and stiffness is the primary cause. This study explores the technical aspects of endoscopic implantation of fat as an autologous implant for restoring the bulk and pliability of the vocal folds. Fat from the buccal area was harvested and prepared in 6 dogs. A mucosal flap submucosal pocket was prepared by means of microlaryngoscopy instrumentation. The fat implant was placed into the submucosal pocket. The mucosal flap was sutured endoscopically. Six weeks later, the larynx was harvested and mounted, and vocal fold oscillation was studied while driven by a humidified flow source. The vibratory patterns were studied by stroboscopy. Histologic sections of the vocal folds were made in the coronal plane. All 6 specimens had histologic evidence of viable implanted fat and/or fibrous tissue at the implant site. The site of implantation was in the superior aspect of the vocal fold, but contributed to mass in vocal fold bulk. This increase in bulk histologically corresponded to stroboscopic evidence of increased mass. On stroboscopy, the implant side continued to demonstrate good vibratory function. The study shows that fat implantation can be carried out as an endoscopic procedure. Fat implantation may be useful as a surgical procedure for restoration of Reinke's space. It may be applicable in patients with scars, sulcus vocalis, or vocal fold atrophy. PMID- 10453781 TI - Inner ear damage in TORP-operated ears: experimental study on danger from environmental air pressure changes. AB - In some cases of sudden inner ear hearing loss in ears with a total ossicular replacement prosthesis, the prosthesis has been found to be penetrating the footplate. Some authors have assumed an external pressure increase for this event. In this study I tried to estimate experimentally the pressure needed for perforating a normal footplate in a temporal bone model. From the data I concluded that "cracking" the footplate can hardly be due to 1 major event of increased pressure. On the other hand, different mechanisms making the footplate a "weak spot" must be discussed and further investigated. In those cases, secondary to thinned or even perforated footplates, environmental pressure changes may be dangerous. PMID- 10453782 TI - Mastoid buffering properties: I. Gas partial pressures. AB - Differences in the gas partial pressures between the middle ear (ME) cavity and the blood are an important factor in ME gas economy. Differences in gas partial pressures between various regions of the ME-mastoid air cell system (ME-MACS) could play a role as well. To determine whether gas partial pressure differences do occur between various compartments in the ME, we measured the rate of gas diffusion from one compartment to another in both an artificial model and in the ME-MACS of human temporal bones. The rate of gas diffusion between various areas of the ME and the mastoid tip was found to be rapid, with a half-life on the order of 2 minutes (range 0.8 to 5.3 minutes). We suggest that this high diffusion rate prevents the buildup of significant differences in gas composition in the ME-MACS system, which can therefore be regarded as a homogeneous gas pocket. PMID- 10453783 TI - Inflammatory response to chronic otitis media in DiGeorge syndrome: a case study using immunohistochemistry on archival temporal bone sections. AB - Immunohistochemical analyses were conducted on archival celloidin-embedded human temporal bone sections from an 8-month-old boy with chronic otitis media and DiGeorge syndrome. We employed antigen retrieval methods with saturated sodium hydroxide-methanol solution, microwave incubation, and proteolytic treatment to demonstrate the distribution of T-lymphocytes, B-lymphocytes, macrophages, and intercellular adhesion molecule 1 (ICAM-1) expression in the middle ear. B lymphocytes and macrophages were observed predominantly within the middle ear mucosa. T-lymphocytes were rare. Further, ICAM-1 was expressed in the vascular endothelium of the lamina propria, as well as infiltrating mononuclear cells. This suggests that the expression of ICAM-1 can be induced in the middle ear with otitis media, even if T-lymphocytes are depressed in a cell-mediated immunodeficiency disorder such as DiGeorge syndrome. PMID- 10453784 TI - Mucin gene expression in the rat middle ear: an improved method for RNA harvest. AB - Mucins are heavily glycosylated proteins characterized by high molecular weight and heterogeneous structure. Mucin genes are expressed in a tissue- or epithelium specific manner. Although mucins are known to be important structural components of the mucociliary transport system that protects epithelium against invading microorganisms, very little is known about mucin gene expression unique to the middle ear. This study demonstrated that middle ear messenger RNA specifically hybridized with rat MUC2 and human MUC2 (SMUC-41) complementary DNA probes. MUC3 and MUC5AC mucin genes, dominantly expressed in rodent intestine and trachea, were not detected in the rat middle ears in this study. The middle ear MUC2 messenger RNA harvested by lavage was characterized by a single transcript- unlike its counterpart in intestine and airways, which is characterized by polydispersity--suggestive of a better method for RNA analysis. It was concluded that rat middle ears possess a MUC2 mucin gene or homologue of human MUC2 (SMUC 41). PMID- 10453785 TI - Immunohistochemical distribution of extracellular matrix components and keratin in experimentally induced otitis media. AB - The distribution of collagen types I, III, and IV and of laminin, fibronectin, and keratin was studied in otitis media experimentally induced by Streptococcus pneumoniae in the chinchilla. The expression of interstitial collagen types I and III and of fibronectin was increased in the subepithelial space that was thickened by inflammation in the acute period of infection. The expression of collagen type IV in the subepithelial space could be seen in the early period. The epithelial cells in the middle ear changed from flat cuboidal to pseudostratified columnar in pneumococcus-inoculated ears, and the number of keratin-positive epithelial cells in the middle ear increased remarkably after infection. These results indicate that changes in epithelial cell differentiation effected by the extracellular matrix correlate with changes in expression of keratin. It is proposed that the extracellular matrix may contribute to tissue repair in the middle ear after bacterial infection by interfering with cell proliferation of epithelial cells and fibroblasts. PMID- 10453786 TI - Ankle-arm index as a screening examination for fibula free tissue transfer. AB - The frequent use of the fibula free flap for mandibular reconstruction has served as an impetus to develop a screening test for this flap. This would detect lower leg vascular disease that could jeopardize the flap or donor extremity. Data would allow a decision whether to pursue further lower leg imaging versus consideration of an alternative donor site. The ankle-arm index (AAI) was determined on a prospective series of 62 patients being considered for fibula free flap transfer. An AAI of less than 1.0 was found to be predictive of a high rate of lower leg vascular disease (by color flow Doppler imaging) that would jeopardize the flap or donor extremity. The AAI did not detect anomalous vasculature of the lower leg (N = 2). If the AAI is less than 1, an alternative donor site to the fibula should be considered, and if it is greater than 1, a color flow Doppler examination of the donor extremity is recommended. PMID- 10453787 TI - Nitric oxide synthase type 3 is increased in squamous hyperplasia, dysplasia, and squamous cell carcinoma of the head and neck. AB - The implication of nitric oxide (NO*) in the multistep process of carcinogenesis prompted us to examine the expression of endothelial constitutive nitric oxide synthase (NOS3) in head and neck squamous cell carcinoma (HNSCCa). Eleven paraffin-embedded samples of normal oral mucosa, 3 reactive oral lesions, 13 samples of squamous dysplasia, and 120 specimens of HNSCCa were immunostained with an anti-NOS3 monoclonal antibody and graded on a 0 to 4+ scale of intensity. Normal squamous mucosa demonstrated very little NOS3 expression. Areas of normal mucosa, reactive mucosa, and dysplastic lesions associated with inflammation tended to demonstrate regional expression of NOS3. Reactive mucosal lesions, squamous dysplasia, and HNSCCa demonstrated a significant (p<.0001) increase in global expression of NOS3. Therefore, NOS3 is expressed very little in histologically normal squamous mucosa, while squamous hyperplasia, dysplasia, and HNSCCa express significantly more NOS3. Regional variation in NOS3 expression appears to be associated with perilesional inflammation. PMID- 10453788 TI - Correcting vocal fold immobility by autologous collagen injection for voice rehabilitation. A short-term study. AB - We report on a short-term clinical study of injectable autologous collagen (Vocalogen) used to correct dysphonias arising from vocal fold immobility. The collagen is extracted from skin taken from the lower abdominal quadrant area or from just above the bikini line. About 30 cm2 of skin are required to provide 2 mL of injectable collagen. The histologic examination of the preparation before injection disclosed the presence of elastin fibers and some clusters of epithelial cells, beside the collagen fibers. The collagen is naturally reticulated, and the molecule is preserved in its entirety. The technique is exactly the same as that reported for bovine collagen: injection into Reinke's space, under general anesthesia, monitored by direct microlaryngoscopy. The amount injected is also similar: 1.5 mL for correction of glottic insufficiency in which the immobile vocal fold is in the intermediate position. Eight patients were injected, and the average follow-up was 4.5 months. Voice assessments made before and after the treatment included stroboscopy, subjective and perceptual judgments, and aerodynamic and acoustic measurements. The functional results were similar to those obtained with bovine collagen. No complications arose. The probability of any hypersensitivity reaction, always a possibility to be feared with bovine collagen, is negligible with the autologous collagen. Long-term results are as yet unknown, but from the fact that the collagen molecular structure is intact and there is little risk of foreign body response, it would be expected that autologous preparations would be more stable than bovine collagen; this appears to be the case in cosmetic applications. Autologous collagen could be employed for the same indications as bovine collagen, provided that a delay of 45 days (the time required to prepare the injectable collagen) is acceptable. The amount of collagen required is also a limiting factor, since the patient's own skin is the starting donor material. PMID- 10453789 TI - Leiomyosarcoma of the larynx. Case report. AB - Leiomyosarcoma involving the larynx is extremely rare and may be difficult to diagnose. Likewise, because of the rarity of this lesion, little information exists with regard to long-term follow-up or optimal management. Reported here is a review of the literature and a case report of a patient with leiomyosarcoma of the larynx treated by surgery alone with 5 years of follow-up. PMID- 10453790 TI - Quantitative glycohistochemical characterization of normal nasal mucosa, and of single as opposed to massive nasal polyps. AB - A series of 41 nasal polyps (23 single and 18 massive) and 6 normal nasal mucosa specimens was glycohistochemically investigated. Five plant lectins were used, i.e., the peanut agglutinin (PNA), the wheat germ agglutinin (WGA), the gorse seed agglutinin (UEA-I), the Maackia amurensis agglutinin (MAA), and the elderberry bark agglutinin (SNA). A neoglycoconjugate and 2 animal lectins (CL-14 and CL-16) were also used. Three quantitative features were calculated by means of computer-assisted microscopy: the percentage of tissue area specifically stained by the histochemical probe, the staining intensity, and the heterogeneity level of the staining distribution. The results show that with respect to sialic acid-glycoprotein binding characteristics as determined by SNA, MAA, and WGA probes, the normal nasal mucosa differed markedly (p<.00001) from the polyposal one. The single nasal polyps exhibited glycohistochemical characteristics that differed markedly (p = .0004) from those exhibited by the massive ones. These differences related mainly to the UEA-I, PNA, and the Thomsen-Friedenreich antigen-exposing neoglycoprotein binding characteristics. In conclusion, the present study shows unambiguously that polyposal mucosa, whether of the single or the massive type, exhibits markedly distinct glycohistochemical characteristics when compared to normal nasal mucosa, and that single nasal polyps also differ markedly from massive ones. PMID- 10453791 TI - Esophageal contractility synchronous with expiration. AB - On the basis of our previous finding that the cervical esophagus was closed during tracheoesophageal phonation, we postulated that the muscle of the cervical esophagus actually contracted during expiration and speculated on its possible regulation by the recurrent laryngeal nerve. Electromyography of the esophageal musculature and fluoroscopy of the esophagus were performed in laryngectomees to demonstrate whether the esophagus contracted during expiration or not. Electromyography, performed in 2 subjects, revealed a burst of discharges synchronous with expiration. Fluoroscopy during tracheoesophageal phonation was performed in 13 subjects to investigate the influence of resection of the esophageal branch of the recurrent nerve on esophageal motor activity. Among 13 subjects, 4 subjects who underwent paratracheal dissection (PTD) at the time of laryngectomy showed a lower superior limit of esophageal closure than did the 9 subjects without PTD, indicating that the cervical esophagus in PTD loses its contractility with the sacrifice of the esophageal branch of the recurrent nerve. We concluded that the cervical esophagus is closed by muscle contraction synchronous with expiration, preventing air entry into the stomach during deep expiration or phonation, and that the esophageal branch of the recurrent nerve is involved. PMID- 10453792 TI - Endoscopic diverticulotomy for the treatment of Zenker's diverticulum: results in 102 patients with staple-assisted endoscopy. AB - Endoscopic diverticulotomy for the treatment of Zenker's diverticulum has been reported infrequently in the literature and has engendered considerable controversy. Between March 1992 and September 1996, we attempted to treat 102 patients with endoscopic treatment for pharyngoesophageal diverticula. In 98 patients, the endoscopic surgery was successfully completed. Conversion to open surgery was required in 4 patients (3.92%). One cartridge of staples in 16 patients (16.32%), 2 cartridges in 78 patients (79.59%), and 3 cartridges in 4 patients (4.08%) were used, according to the size of the diverticulum; the median duration of the procedure was 20 minutes (10 to 60 minutes). No postoperative morbidity or mortality was recorded. Oral feeding was started following radiologic control after a median of 2 days; the median hospital stay was 4 days. The median follow-up is 16 months (1 to 45 months). Four patients operated on before the introduction of the modified stapler showed a persistent diverticular pouch: 3 underwent repeat endoscopic operation, and 1 underwent conventional open surgery. All treated patients are asymptomatic. Manometric study performed in 15 patients showed a significant reduction of basal upper esophageal sphincter pressure compared to preoperative data (48.30+/-21.74 versus 29.38+/-5.68 mm Hg; p<.01). We therefore recommend endoscopic diverticulotomy, considering that the procedure is relatively safe and effective, with minimal patient discomfort, and the results are equal to those of the external approach. This procedure offers the advantages of short hospitalization, rapid convalescence, brief operative time, absence of skin incision. predictable resolution of symptoms, and reduced morbidity. PMID- 10453793 TI - High-resolution magnetic resonance imaging of recurrent pneumoparotitis. PMID- 10453794 TI - Antipsychotic polypharmacy, Part 1: Therapeutic option or dirty little secret? AB - Antipsychotic polypharmacy is a surprisingly frequent occurrence that can be both justified and unjustifed, depending on how it is used. To the extent that this phenomenon has been unrecognized and is not being studied, it is a "dirty little secret." To the extent that careful clinicians have uncovered a useful strategy for boosting the effectiveness of available antipsychotic monotherapies, it represents an opportunity to improve the outcomes of patients with psychotic illnesses. PMID- 10453795 TI - The economic burden of anxiety disorders in the 1990s. AB - BACKGROUND: We assess the annual economic burden of anxiety disorders in the United States from a societal perspective. METHOD: Using data from the National Comorbidity Study, we applied multivariate regression techniques to calculate the costs associated with anxiety disorders, after adjusting for demographic characteristics and the presence of comorbid psychiatric conditions. Based on additional data, in part from a large managed care organization, we estimated a human capital model of the societal cost of anxiety disorders. RESULTS: We estimated the annual cost of anxiety disorders to be approximately $42.3 billion in 1990 in the United States, or $1542 per sufferer. This comprises $23.0 billion (or 54% of the total cost) in nonpsychiatric medical treatment costs, S13.3 billion (31%) in psychiatric treatment costs, $4.1 billion (10%) in indirect workplace costs, $1.2 billion (3%) in mortality costs, and $0.8 billion (2%) in prescription pharmaceutical costs. Of the $256 in workplace costs per anxious worker, 88% is attributable to lost productivity while at work as opposed to absenteeism. Posttraumatic stress disorder and panic disorder are the anxiety disorders found to have the highest rates of service use. Other than simple phobia, all anxiety disorders analyzed are associated with impairment in workplace performance. CONCLUSION: Anxiety disorders impose a substantial cost on society, much of which may be avoidable with more widespread awareness, recognition, and appropriate early intervention. PMID- 10453796 TI - Development of depression during placebo-controlled trials of bupropion for smoking cessation: case reports. AB - BACKGROUND: Recent attention has focused on the relationship between depression and smoking cessation. This article describes 5 cases of severe depression that occurred during 2 multicenter trials using bupropion for smoking cessation. METHOD: Subjects were participants in 2 randomized, double-blind, placebo controlled studies investigating the efficacy of bupropion for smoking cessation. Data from both trials were restricted to subjects at the Rochester, Minn., site in order to have access to the medical records for information on depression diagnosis, treatment, and follow-up. The first trial involved 205 smokers who received active bupropion or placebo for 7 weeks. In the second trial, 252 smokers received open-label bupropion therapy for 7 weeks. Those abstinent from smoking at the end of week 7 (N = 148) were randomly assigned to a 45-week, double-blind, relapse-prevention phase. RESULTS: In the first trial, 1 of the 205 participants (0.49%) experienced major depression during the 7-week treatment phase. In the second trial, none of the 252 subjects developed major depression during the 7-week, open-label phase. When results of both trials across the 7 week treatment phase (study 1, N = 205; study 2, N = 252) are combined, the rate of developing major depression was 0.22% (1 of 457). Of the 457 subjects, none of the 51 who received placebo and 1 (0.25%) of the 406 who received active bupropion developed major depression. In the second trial, 4 (2.7%) of the 148 subjects randomly assigned to the 45-week, relapse-prevention phase developed depression. Overall, 4 of the 5 cases from the 2 trials had a past history of major depression prior to study entry, but none had current major depression. CONCLUSION: Major depression may occur in some individuals during smoking cessation treatment with bupropion. PMID- 10453797 TI - Open trial of flutamide for treatment of obsessive-compulsive disorder. AB - BACKGROUND: Several lines of evidence suggest that gonadal steroid hormones play a role in the onset and exacerbation of obsessive-compulsive disorder (OCD). In this study, we examined the effects of treatment with flutamide, a synthetic, nonsteroidal, competitive antagonist of the androgen receptor, on OCD symptoms. METHOD: Eight outpatients meeting DSM-III-R criteria for OCD participated in an 8 week open trial of flutamide. The dose was increased from 250 mg/day to 750 mg/day over the first 4 weeks and maintained at 750 mg/day for the final 4 weeks. The primary outcome measures for OCD symptoms were the Yale-Brown Obsessive Compulsive Scale and the Maudsley Inventory and for anxiety symptoms, the Beck Anxiety Inventory and the Hamilton Rating Scale for Anxiety. Subjects also provided self-ratings of aggression and sexual interest and activity. RESULTS: There were no reductions in measures of obsession and compulsions or measures of anxiety over the 8-week trial. However, self-ratings of feelings of aggression did fall significantly over the 8-week trial (p < .001). CONCLUSION: The lack of response to treatment with flutamide, an androgen receptor antagonist, suggests that any effects of gonadal steroids to exacerbate OCD symptoms are more likely to be mediated through estrogen receptors or through mechanisms that do not involve classical intracellular androgen receptors. Future treatment trials should examine agents that antagonize estrogen receptors or otherwise inhibit estrogen activity. PMID- 10453798 TI - Mirtazapine in major depression with comorbid generalized anxiety disorder. AB - BACKGROUND: A high proportion of patients with generalized anxiety disorder (GAD) have comorbid depressive illness. The presence of anxiety in depression has significant prognostic implications. Because of mirtazapine's early anxiolytic effects, the present study was undertaken as a preliminary investigation in patients with a diagnosis of major depression with comorbid GAD. METHOD: Mirtazapine was administered to 10 patients with DSM-IV major depressive disorder and comorbid GAD in an 8-week open-label study. Mirtazapine was increased from an initial daily dose of 15 mg to a maximum daily dose of 45 mg. RESULTS: Patients were found to have significant reductions in Hamilton Rating Scale for Depression scores, Hamilton Rating Scale for Anxiety scores, and Beck Depression Inventory scores, with improvement noted after the first week of therapy and continuing improvement over the 8 weeks of study. CONCLUSION: These positive preliminary findings support the further investigation of mirtazapine's potential value as a treatment for generalized anxiety disorder in addition to its established efficacy as an antidepressant drug. PMID- 10453799 TI - Measurement of compliance with naltrexone in the treatment of alcohol dependence: research and clinical implications. AB - BACKGROUND: Medication compliance is a critical issue in pharmacotherapy. This study evaluated the clinical utility of the Medication Event Monitoring System (MEMS), a newer method for monitoring medication compliance, compared with pill count, a traditional measure, in a sample of patients treated for alcohol dependence with naltrexone. METHOD: Ninety-three outpatients meeting DSM-III-R criteria for alcohol dependence participated in a 10-week open-label study of naltrexone. They were provided with naltrexone, 50 mg daily, and concurrent counseling. Measures of medication compliance and drinking during treatment were collected every 2 weeks. RESULTS: Pill count yielded a significantly (p < .001) higher estimate of compliance (87.6%+/-18.1%) than the MEMS (80.4%+/-20.6%). However, the estimate of compliance obtained with the MEMS was more consistently correlated with treatment outcome (percentage of days abstinent, percentage of heavy drinking days, and mean alcohol amount consumed per drinking occasion) than the pill count compliance rate. In addition, classification of the sample into compliant and less compliant groups using the MEMS data yielded groups that differed more clearly on drinking outcomes than did stratification on the basis of pill count. CONCLUSION: In pharmacotherapy research, the MEMS may provide more reliable and valid information about subjects' medication compliance than pill count. Clinically, information obtained with the MEMS could be used to provide feedback to patients about their pill-taking behavior to enhance compliance and overall outcome of therapy. PMID- 10453800 TI - Self-injurious skin picking: clinical characteristics and comorbidity. AB - BACKGROUND: Repetitive skin picking, a self-injurious behavior that may cause severe tissue damage, has received scant empirical attention. The authors examined the demographics, phenomenology, and associated psychopathology in a series of 31 subjects with this problem. METHOD: Subjects were administered the Structured Clinical Interview for DSM-IV for Axis I and Axis II disorders. They also completed several mood questionnaires and a new self-report inventory designed to assess phenomenology, triggers, cognitions, emotions, and consequences associated with skin picking. RESULTS: The mean age at onset on self injurious skin picking was 15 years, and the mean duration of illness was 21 years. All subjects picked at more than one body area, and the most frequent sites of skin picking were pimples and scabs (87%). The most common comorbid Axis I diagnoses were obsessive-compulsive disorder (OCD; 52%), alcohol abuse/dependence (39%), and body dysmorphic disorder (32%). Forty-eight percent (N = 15) of the subjects met criteria for at least one mood disorder, and 65% (N = 20) for at least one anxiety disorder. The most common Axis II disorders were obsessive-compulsive personality disorder (48%) and borderline personality disorder (26%). CONCLUSION: Self-injurious skin picking is a severe and chronic psychiatric and dermatologic problem associated with high rates of psychiatric comorbidity. It may be conceptualized as a variant of OCD or impulse-control disorder with self-injurious features and may, in some cases, represent an attempt to regulate intense emotions. PMID- 10453801 TI - Late-onset adrenoleukodystrophy associated with long-standing psychiatric symptoms. AB - BACKGROUND: It is not commonly appreciated that patients with adrenoleukodystrophy (ALD) can first present in adulthood with psychiatric symptoms. METHOD: This case study involved a 31-year-old man who was referred for a neuropsychiatric assessment of tardive dyskinesia and treatment-resistant psychosis. Upon neurologic examination, he was found to have spasticity, marked hyperreflexia with clonus, and bilateral Babinski signs. T2-weighted magnetic resonance imaging demonstrated severe white matter disease. Metabolic screening revealed abnormalities of very long chain fatty acids consistent with the diagnosis of ALD. These results prompted us to review the literature on late onset ALD with attention to (1) the nature of the associated psychiatric and neurologic symptoms, (2) the neuroimaging abnormalities associated with this disorder, and (3) treatment considerations. RESULTS: Individuals with adult-onset ALD may initially present with psychiatric symptomatology. Most commonly, these patients manifest signs of mania including disinhibition, impulsivity, increased spending, hypersexuality, loudness, and perseveration. ALD patients will often have upper motor neuron findings on neurologic examination. Despite the name of the disease, patients with ALD may not have clinical evidence of adrenal dysfunction. Neuroimaging reveals diffuse, confluent white matter lesions that typically originate in the parieto-occipital region. Both neuroleptic and anticholinergic medications may result in significant side effects with little resolution of the underlying psychiatric symptoms. CONCLUSION: This case study and review of the literature illustrate the importance of performing neurologic and radiological examinations on all psychiatric patients with chronic illnesses. We emphasize the importance of reexamining and reimaging patients who are not responding to standard treatment. The clinical problem of "treatment resistance" should be seen as an indication that other diagnoses, such as an underlying metabolic disorder, need to be considered. PMID- 10453802 TI - A pilot study on risperidone metabolism: the role of cytochromes P450 2D6 and 3A. AB - BACKGROUND: The limited available information on plasma risperidone levels shows a stable relationship between daily doses of risperidone and total plasma concentration (risperidone plus its active metabolite 9-hydroxyrisperidone). The ratio between risperidone and 9-hydroxyrisperidone characterizes cytochrome P450 2D6 (CYP2D6) status. According to the manufacturer, the CYP2D6 genotype or drugs that influence CYP2D6 or other cytochrome P450 isoenzyme activity are not expected to be clinically significant. One case report suggests that CYP3A participates in the metabolism of risperidone. METHOD: A case series of 13 risperidone patients (the initial case and 12 new cases) who were genotyped for CYP2D6 were followed, and another 20 risperidone patients from a case-control study for the CYP2D6 genotype were reviewed. RESULTS: The CYP2D6 poor metabolizers, who are enzyme deficient (2/13 in the case series and 3/20 in the case-control study), did not appear to tolerate risperidone well. Drugs affecting CYP3A, in particular powerful inducers and inhibitors, resulted in at least a 2 fold decrease or increase in plasma risperidone levels. CONCLUSION: The anecdotal nature of this study is clearly a limitation. Drugs influencing CYP3A and CYP2D6 metabolic activity may significantly affect risperidone levels. Thus, plasma level monitoring of risperidone in a clinical setting may be useful, especially if patients are taking multiple medications or a CYP2D6 deficiency is suspected. New prospective studies under more controlled conditions are needed to verify these hypotheses. PMID- 10453803 TI - Clozapine reduces severe self-mutilation and aggression in psychotic patients with borderline personality disorder. AB - BACKGROUND: Clozapine has been reported to be effective in diminishing violence toward others in psychotic patients. This article describes the impact of clozapine on severe self-mutilation among patients with the dual diagnoses of borderline personality disorder and persistent psychoses. METHOD: Seven subjects known to the authors were selected for careful chart audits. These subjects had been admitted to 2 state psychiatric hospitals owing to severe self-mutilation and/or violence and subsequently treated with clozapine. A mirror-image design anchored to the start date of clozapine treatment and extending in either direction to a maximum of 1 year was used to extract data. Data extracted included incidents of self-mutilation (restraint), seclusion, the as and when needed (p.r.n.) use of medications, injuries to staff and peers, hospital privileges, and Global Assessment of Functioning (GAF) scores. RESULTS: The subjects were all white women with a mean age of 37 years. All subjects carried DSM-III-R or DSM-IV borderline personality disorder diagnoses and an Axis I disorder diagnosis. They had received trials of several psychotropic agents, often in combination and mostly without benefit. After clozapine treatment, there were statistically significant reductions in incidents of self-mutilation (restraint), seclusion, the use of p.r.n. antianxiety medications, and injuries to staff and peers. These subjects received higher levels of hospital privileges, and their GAF scores nearly doubled following clozapine treatment. Four subjects were subsequently discharged from hospital. CONCLUSION: These preliminary but nonetheless favorable results suggest that clozapine deserves careful consideration for a controlled study in patients with borderline personality disorder and psychoses, especially if the clinical issues include severe self mutilation, aggression, and violence. Until such studies are done, the risk-to benefit ratio of clozapine treatment needs to be carefully evaluated on an individualized basis in such subjects. PMID- 10453804 TI - Effects of "Ecstasy" blocked by serotonin reuptake inhibitors. PMID- 10453805 TI - Chorea and tardive dyskinesia in a patient taking risperidone. PMID- 10453806 TI - Clozapine versus risperidone in treatment-refractory schizophrenia: possible impact of dosing strategies. PMID- 10453807 TI - Panic disorder in the primary care setting: comorbidity, disability, service utilization, and treatment. AB - BACKGROUND: Increased medical service utilization in patients with panic disorder has been described in epidemiologic studies, although service use in primary care panic patients relative to other primary care patients is less well characterized. Inadequate recognition of panic has been shown in several primary care studies, although the nature of usual care for panic in this setting has not been well documented. This study aimed to document increased service use in panic patients relative to other primary care patients and to characterize the nature of their usual care for panic and their outcome. METHOD: Using a waiting room screening questionnaire and follow-up telephone interview with the Composite International Diagnostic Interview, we identified a convenience sample of 81 patients with panic disorder (DSM-IV) and a control group of 183 psychiatrically healthy patients in 3 primary care settings on the West Coast and determined psychiatric diagnostic comorbidity, panic characteristics, disability, and medical and mental health service use, including medications. A subsample (N = 41) of panic patients was reinterviewed 4-10 months later to determine the persistence of panic and the adequacy of intervening treatment received using the Harvard/Brown Anxiety Disorders Research Program study criteria for cognitive behavioral therapy (CBT) and an algorithm developed by the authors for medications. RESULTS: Seventy percent of panic patients had a comorbid psychiatric diagnosis. Patients had more disability in the last month (days missed or cut down activities) (p < .01), more utilization of emergency room and medical provider visits (p < .01), and more mental health visits (p < .05). Despite the latter, only 42% received psychotropic medication, 36% psychotherapy, and 64% any treatment. On follow-up, 85% still met diagnostic criteria for panic, and only 22% had received adequate medication (type and/or dose) and 12% adequate (i.e., CBT) psychotherapy. CONCLUSION: These findings suggest a need for improved treatment interventions for panic disorder in the primary care setting to decrease disability and potentially inappropriate medical service utilization. PMID- 10453808 TI - Invited commentary: propensity scores. AB - The propensity score is the conditional probability of exposure to a treatment given observed covariates. In a cohort study, matching or stratifying treated and control subjects on a single variable, the propensity score, tends to balance all of the observed covariates; however, unlike random assignment of treatments, the propensity score may not also balance unobserved covariates. The authors review the uses and limitations of propensity scores and provide a brief outline of associated statistical theory. They also present a new result of using propensity scores in case-cohort studies. PMID- 10453809 TI - Recent alcohol intake as estimated by the Health Habits and History Questionnaire, the Harvard Semiquantitative Food Frequency Questionnaire, and a more detailed alcohol intake questionnaire. AB - Epidemiologic studies often rely on food frequency questionnaires (FFQs) to collect information on alcoholic beverage intake. However, estimation of alcohol intake using FFQs may be of some concern because of limited questions concerning alcohol intake. The authors compared estimates of alcohol intake during the 12-24 months prior to interview obtained from the Health Habits and History Questionnaire and the Harvard Semiquantitative Food Frequency Questionnaire with those from a more extensive alcohol questionnaire, the Drinking Pattern Questionnaire, among 133 healthy subjects (75 men, 58 women) aged 35-73 years, residents of western New York State. Data were collected in 1995 during two separate interviewer-administered computer-assisted interviews conducted approximately 2 weeks apart. For each questionnaire, average daily ounces (1 oz = 30 ml) of alcohol intake from alcoholic beverages were calculated as the product of the reported beverage-specific drink size (ounces) and the average daily frequency of intake multiplied by a factor representing the percentage of alcohol provided by each beverage. Estimates of total alcohol and liquor intake, but not of beer and wine intake, tended to be higher for the Drinking Pattern Questionnaire compared with the FFQs. Spearman's correlation coefficients ranged from 0.69 to 0.84. These results suggest that although the Drinking Pattern Questionnaire produced higher estimates than either FFQ, both FFQs provide a reasonable ranking of participants' alcohol intake. PMID- 10453810 TI - Underestimation of risk associations due to regression dilution in long-term follow-up of prospective studies. AB - In prospective studies, disease rates during follow-up are typically analyzed with respect to the values of factors measured during an initial baseline survey. However, because of "regression dilution," this generally tends to underestimate the real associations of disease rates with the "usual" levels of such risk factors during some particular exposure period. The "regression dilution ratio" describes the ratio of the steepness of the uncorrected association to that of the real association. To assess the relevance of the usual value of a risk factor during particular exposure periods (e.g., first, second, and third decades) to disease risks, regression dilution ratios can be derived by relating baseline measurements of the risk factor to replicate measurements from a reasonably representative sample of study participants after an interval equivalent to about the midpoint of each exposure period (e.g., at 5, 15, and 25 years, respectively). This report illustrates the impact of this time interval on the magnitude of the regression dilution ratios for blood pressure and blood cholesterol. The analyses were based on biennial remeasurements over 30 years for participants in the Framingham Study (Framingham, Massachusetts) and a 26-year resurvey for a sample of men in the Whitehall Study (London, England). They show that uncorrected associations of disease risk with baseline measurements underestimate the strength of the real associations with usual levels of these risk factors during the first decade of exposure by about one-third, the second decade by about one-half, and the third decade by about two-thirds. Hence, to correct appropriately for regression dilution, replicate measurements of such risk factors may be required at varying intervals after baseline for at least a sample of participants. PMID- 10453811 TI - Evaluation of a population roster as a source of population controls: the Massachusetts Resident Lists. AB - Published population rosters can serve as a convenient source of population controls. The authors evaluated one such roster, the Massachusetts Resident Lists, by estimating the completeness of the Lists and by describing the differences between persons included and not included on the Lists. The subjects were cases from three case-control studies of ovarian cancer conducted in eastern Massachusetts between 1978 and 1996. For each of the three case series, more than 90% of the cases were located on the Resident Lists. Age was one of the primary differences to emerge between cases included and not included; in the most recent case series, cases younger than age 40 years were less likely than older cases to be included on the Lists. PMID- 10453812 TI - Birth-cohort phenomenon in the time trends of mortality from ulcerative colitis. AB - It has been suggested that the mortality trends of ulcerative colitis in England and Wales are shaped by an underlying birth-cohort phenomenon. This pattern implies that exposure to an environmental risk factor early in life plays a crucial role in the development of the disease. The authors tested whether the birth-cohort pattern is unique to British mortality statistics or a common feature of ulcerative colitis in western countries by using the vital statistics from England and Wales, Canada, Scotland, Switzerland, The Netherlands, and the United States. Ulcerative colitis death rates from the six countries were plotted against the periods of death or periods of birth. Mortality from ulcerative colitis increased in successive generations born throughout the second half of the 19th century. It peaked in subjects born between 1880 and 1890 and has declined since then. Strikingly similar patterns were found in the six countries and when women and men were analyzed separately. The birth-cohort pattern indicates that development of ulcerative colitis is strongly influenced by one or several environmental risk factors, which act during a short period early in life. In western countries, exposure to this risk has changed in a similar fashion. PMID- 10453813 TI - Plasma selenium levels and risk of hepatocellular carcinoma among men with chronic hepatitis virus infection. AB - Both experimental and epidemiologic studies have linked a low dietary intake of selenium with an increased risk of cancer. The authors examined the association between plasma selenium levels and risk of hepatocellular carcinoma (HCC) among chronic carriers of hepatitis B and/or C virus in a cohort of 7,342 men in Taiwan who were recruited by personal interview and blood draw during 1988-1992. After these men were followed up for an average of 5.3 years, selenium levels in the stored plasma were measured by using hydride atomic absorption spectrometry for 69 incident HCC cases who were positive for hepatitis B surface antigen (HBsAg) and/or antibodies against hepatitis C virus (mostly HBsAg positive) and 139 matched, healthy controls who were HBsAg positive. Mean selenium levels were significantly lower in the HCC cases than in the HBsAg-positive controls (p = 0.01). Adjusted odds ratios of HCC for subjects in increasing quintiles of plasma selenium were 1.00, 0.52, 0.32, 0.19, and 0.62, respectively. The inverse association between plasma selenium levels and HCC was most striking among cigarette smokers and among subjects with low plasma levels of retinol or various carotenoids. There was no clear evidence for an interaction between selenium and alpha-tocopherol in relation to HCC risk. PMID- 10453814 TI - Case-control study of non-Hodgkin's lymphoma among women and heterosexual men in the San Francisco Bay Area, California. AB - A population-based case-control study was conducted between 1988 and 1995 in the San Francisco Bay Area of California to determine risk factors for non-Hodgkin's lymphoma. Participants completed in-person interviews, and blood was drawn to test for viruses and lymphocyte subsets. This report includes data for 1,281 cases and 2,095 controls. In multivariate analyses, the factors associated with a decreased risk for non-Hodgkin's lymphoma were allergy to plants, bee and wasp stings, five or more vaccinations, drugs to lower blood cholesterol, nonsteroidal anti-inflammatory drugs, total number of sexual partners, and lifetime marijuana use, whereas an increased risk was associated with cimetidine and other histamine H2-receptor antagonists, splenectomy, gonorrhea, and body mass index. Unique to sex-specific models was an increased risk for endocrine gland disorders among women and for polio among men. Median CD3, CD4, CD8, CD20, and lymphocyte counts for non-Hodgkin's lymphoma patients were significantly lower than those for controls. These results implicate environmental factors that may influence the early stages of lymphomagenesis by stimulating the immune system. Antigen-driven B cells that accumulate to form lymphoma may be suppressed by immunologic stresses such as exposure to an increased number of sexual partners and to certain medications. A history of allergies provides evidence for a persistent capacity for B-cell differentiation and therefore a decreased accumulation of B cells. The decreased risk for non-Hodgkin's lymphoma with use of nonsteroidal anti-inflammatory drugs and cholesterol-lowering drugs is consistent with a macrophage inflammatory role in B-cell proliferation. PMID- 10453815 TI - Body mass index and colon cancer in a national sample of adult US men and women. AB - The evidence supporting obesity as a risk factor for colon cancer remains inconclusive, especially among women. The author studied the association between obesity and colon cancer in a nationally representative cohort of men and women aged 25-74 years who participated in the First National Health and Nutrition Examination Survey from 1971 to 1975 and were subsequently followed up through 1992. Among the 13,420 persons included in the analytic sample, 222 incident cases of colon cancer were identified. Height and weight were measured during the baseline examination. Compared with participants whose body mass index was less than 22 kg/m2, the hazard ratios were 1.79 (95% confidence interval (CI): 0.87, 3.71), 1.86 (95% CI: 0.86, 4.03), 2.47 (95% CI: 1.14, 5.32), 3.72 (95% CI: 1.68, 8.22), and 2.79 (95% CI: 1.22, 6.35) for participants with a body mass index of 22-<24 kg/m2, 24-<26 kg/m2, 26-<28 kg/m2, 28-<30 kg/m2, and > or = 30 kg/m2, respectively. The hazard ratios were similar for men and women. Subscapular skinfold thickness, but not triceps skinfold thickness, was positively associated with colon cancer incidence among men but not women, after adjustment for body mass index and other possible confounders. These results strongly support the hypothesis that excess body weight is a risk factor for colon cancer among both men and women. PMID- 10453816 TI - Consequences of the use of different measures of effect to determine the impact of age on the association between obesity and mortality. AB - The impact of using different measures of the effect of obesity on mortality across age groups has not been explored. The authors examined this issue by using mortality data from 62,116 men and 262,965 women in the Cancer Prevention Study-I (1960-1972). Measures of effect were calculated separately, by decade of age, for five groups of participants aged 30-79 years. The rate ratio associated with obesity declined with age, from 2.60 (men) and 1.99 (women) for participants aged 30-39 years to 1.24 (men) and 1.15 (women) for those aged 70-79 years. In contrast, the rate differences between obese and reference-weight participants increased with age, from 201 (men) and 112 (women) deaths per 100,000 person years for those in the youngest decade to 1,379 (men) and 626 (women) deaths per 100,000 person-years for those in the oldest decade. The years of life lost attributable to obesity tended to increase with age but declined for those in the oldest decade. The rate advancement period declined with decade of age. Standardization of estimates to a population changed some age-associated trends. The direction of trends varied regarding the effect of obesity on mortality across age groups, depending on the measure of effect used. PMID- 10453817 TI - Physical activity and cardiovascular disease risk in middle-aged and older women. AB - The authors investigated the relation between physical activity and cardiovascular disease (CVD) in women by following 1,564 University of Pennsylvania alumnae (mean age, 45.5 years), initially free of CVD, from 1962 until 1993. Energy expenditure was estimated from the daily number of flights of stairs climbed and blocks walked as well as the sports played and was categorized into approximate thirds (<500, 500-999, > or = 1,000 kcal/week). During 35,021 person-years, 181 CVD cases were identified. After adjustment for coronary risk factors, the relative risks of CVD were 0.99 (95% confidence interval (CI): 0.69, 1.41) and 0.88 (95% CI: 0.62, 1.25) for women who expended 500-999 and > or = 1,000, respectively, compared with <500 kcal/week (p for trend = 0.45). Only walking was found to be inversely related to CVD risk (p for trend = 0.054). Compared with women who walked <4 blocks/day, the relative risks of CVD were 0.84 (95% CI: 0.59, 1.19) and 0.67 (95% CI: 0.45, 1.01) for women who walked 4-9 and > or = 10 blocks/day, respectively. Finally, an interaction (p = 0.023) between body mass index and physical activity on CVD risk was observed, with an inverse association only for leaner (<23 kg/m2) women. These data showed no overall association of physical activity with CVD risk in women. However, walking > or = 10 blocks/day (approximately 6 miles (9.7 km)/week) was associated with a 33% decreased risk. One explanation for this finding may be that walking was reported more precisely than other kinds of activities. PMID- 10453818 TI - Domestic violence in northern India. AB - This study examined the prevalence and characteristics of wife abuse as reported by nearly 6,700 married men living in five districts of northern India during 1995-1996. In addition, associations between wife abuse and sociodemographic factors were investigated to enable two theoretical/conceptual perspectives regarding abuse to be evaluated: that abuse is more common among families under stress and among more "private" families. The district-specific percentages of men who reported physically abusing their wives ranged from 18% to 45%, with 18 40% of the men in each district having had nonconsensual sex with their wives and 4-9% having physically forced their wives to have sex. The authors used logistic regression analyses to control for a variety of sociodemographic variables and found positive associations between wife abuse and stress-related factors, including the husband having a low educational level, the couple living in poverty, the husband being young when he first lived with his wife, and the couple having multiple children. Contrariwise, there was no strong empirical support for the idea that wife abuse may be more common in more "private" families. PMID- 10453819 TI - Re: "Breastfeeding reduces risk of respiratory illness in infants". PMID- 10453820 TI - Re: "Distinguishing the effects of maternal and offspring genes through studies of 'case-parent triads'" and "a new method for estimating the risk ratio in studies using case-parental control design". PMID- 10453821 TI - Re: "Is there really a heterosexual AIDS epidemic in the United States? Findings from a multisite validation study, 1992-1995". PMID- 10453822 TI - Re: "Weight history, glucose intolerance, and insulin levels in middle-aged Swedish men". PMID- 10453823 TI - The definition of pre-eclampsia. PMID- 10453824 TI - The role of inherited thrombophilia in venous thromboembolism associated with pregnancy. AB - Venous thromboembolism is an important cause of maternal morbidity and mortality. The puerperium should be regarded as the period of greatest risk. However, fatalities in early pregnancy emphasise the need to assess thrombotic risk at all stages of pregnancy. In many cases those at increased risk are potentially identifiable on clinical grounds alone such as those with a personal or family history of venous thromboembolism, obesity, or surgery. Identification of women with multiple clinical risks for thrombosis during pregnancy remains the key to reducing the incidence of this condition. In women who present with a personal or family history of proven venous thromboembolism, thrombophilia screening should be performed in early pregnancy, since the results may influence subsequent management during pregnancy. The investigation and management of patients considered at increased risk of venous thrombosis during pregnancy requires close liaison between obstetricians and haematologists familiar with this rapidly expanding and complex field of thrombophilia. PMID- 10453825 TI - Evaluation of a definition of pre-eclampsia. AB - OBJECTIVES: To determine: 1. whether an alternative definition of gestational hypertension and pre-eclampsia stratifies women according to their risk of maternal and fetal complications; 2. whether pregnancy outcome in women with gestational hypertension differs in the presence or absence of '+' proteinuria; and 3. whether a blood pressure rise of > or = 30/15 mmHg during pregnancy is associated with adverse outcome in women who remain normotensive. DESIGN: Prospective, nested case-control study. SETTING: Community based. POPULATION: Healthy, nulliparous women (n = 1496). METHODS: Women recruited into a study investigating serum markers predictive of pre-eclampsia were classified as having gestational hypertension (systolic blood pressure > or = 140 mmHg with a rise of > or = 30 mmHg and/or diastolic blood pressure > or = 90 mmHg with a rise of > or = 15 mmHg) or pre-eclampsia (gestational hypertension plus proteinuria > or = 2+on dipstick or > 0.3 g/24 h). Maternal and fetal complications in gestational hypertension or pre-eclampsia were compared with a control group of 223 randomly selected normotensive women. The main outcome measures were severe maternal disease, preterm birth and small for gestational age infant. RESULTS: A stepwise increase in adverse maternal and fetal outcomes occurred in gestational hypertension (n = 117, 7.8%) and pre-eclampsia (n = 71, 4.8%). Severe maternal disease developed in 26.5% (21.4% severe hypertension alone, 5.1% multisystem disease) of women with gestational hypertension and 63.4% (21.1% severe hypertension alone, 42.3% multisystem disease) of women with pre-eclampsia (OR 4.8; 95% CI 2.4-9.5). Preterm birth and small for gestational age infants were more frequent in gestational hypertension (OR 1.7; 95% CI 0.5-5.4, and OR 2.0; 95% CI 1.0-3.7, respectively) and pre-eclampsia (OR 14.6; 95% CI 5.8-37.8, and OR 2.6; 95% CI 1.2-5.3) than in the normotensive group. Among women with gestational hypertension severe maternal disease was more common in women with '+' proteinuria (41.7%) than in those with no proteinuria (15.9%): OR 3.8; 95% CI 1.5 9.8. Pregnancies were uncomplicated in the 27% of normotensive women who had a rise of > or = 30 mmHg systolic blood pressure and/or > or = 15 mmHg rise in diastolic blood pressure. CONCLUSIONS: In the nulliparous population studied our definition of gestational hypertension and pre-eclampsia identified women at increasing risk of maternal and fetal complications. In gestational hypertension, the presence of proteinuria '+' was associated with a 3.8-fold increase in severe maternal disease. Normotensive women who have a rise in blood pressure > or = 30/15 mmHg had uncomplicated pregnancies. PMID- 10453826 TI - Antecedents of neonatal encephalopathy with fetal acidaemia at term. AB - OBJECTIVE: To identify the relative contribution of antenatal hypoxia, obstetric catastrophe during labour and fetal monitoring practice to the occurrence of neonatal encephalopathy associated with acidaemia at term. DESIGN: Prospective study. SETTING: Tertiary referral hospital in Auckland, New Zealand. SAMPLE: Twenty-two term babies born between January 1996 and October 1997 with umbilical artery pH < or = 7.09 (median 6.88) or 5 minute Apgar score < 7 (median 5.0), and moderate to severe encephalopathy within five hours of birth. METHODS: Antenatal and intrapartum events and fetal heart rate monitoring practice were reviewed by an experienced obstetrician. RESULTS: More than half the cases were associated with events beyond the control of the clinician: 5 of 22 (23%) had evidence of antenatal hypoxia and 5 of 22 (23%) experienced an obstetric catastrophe during labour. Use of continuous fetal monitoring techniques or the interpretation of fetal heart rate changes was suboptimal in 8 of 12 cases. Continuous monitoring was not performed at all in three cases. All pregnancies were of either low or medium risk; none had proteinuric hypertension and no case was breech, small for gestational age or had a gestational age > or = 42 weeks. CONCLUSIONS: A significant proportion of babies with encephalopathy associated with acidaemia at term experienced either antenatal hypoxia or catastrophic events beyond the control of the clinician. Further improvements in obstetric care will require greater vigilance in low to medium risk pregnancies and improved fetal monitoring practice during both induction and labour. PMID- 10453827 TI - Intrapartum umbilical artery Doppler velocimetry as a predictor of adverse perinatal outcome: a systematic review. AB - OBJECTIVE: To evaluate the diagnostic prediction of intrapartum umbilical artery Doppler velocimetry for adverse perinatal outcomes using systematic quantitative overview of the available literature. DESIGN: Online searching of MEDLINE database (January 1966-September 1997), scanning of bibliography of known primary and review articles, review of recent journal issues and that from personal files. Study selection, assessment of study quality and data extraction were all performed in duplicate under masked conditions. PARTICIPANTS: 2700 women (unselected, low, high, and combined low and high obstetric risk populations) included in eight studies selected for meta-analyses. MAIN OUTCOME MEASURES: Likelihood ratios (LRs) for positive and negative test results were generated for the following outcome measures: Apgar scores < 7 at 1 and 5 minute following delivery, small for gestational age fetus; intrapartum fetal heart rate abnormality, umbilical arterial acidosis at delivery; and caesarean section for fetal distress. RESULTS: For Apgar score < 7 at 1 minute following delivery, the pooled LR was 2.5 (95% CI 1.7-3.7) for a positive test and 1.0 (95% CI 0.9-1.1) for a negative test result. A positive test predicted an Apgar score < 7 at 5 minute following delivery with a pooled LR of 1.3 (95% CI 0.4-4.1) while a negative test had a pooled LR of 1.0 (95% CI 0.8-1.2). For the prediction of a small for gestational age fetus, the pooled LR was 3.4 (95% CI 2.3-5.1) for a positive test and 0.9 (95% CI 0.8-1.0) for a negative test. The prediction for fetal heart rate abnormality during labour was similarly disappointing: the pooled LR for a positive test result was 1.4 (95% CI 0.9-1.2) whereas a negative test result generated a pooled LR of 0.9 (95% CI 0.9-1.0). With umbilical acidosis at delivery, the pooled LR was 1.6 (95% CI 1.1-2.5) for a positive test and 1.1 (95% CI 1.0-1.2) for a negative test. The LRs for the prediction of caesarean section for fetal distress were 4.1 (95% CI 2.7-6.2) for a positive test result and 0.9 (95% CI 0.8-1.0) for a negative test result. CONCLUSION: Intrapartum umbilical artery Doppler velocimetry is a poor predictor of adverse perinatal outcomes. PMID- 10453828 TI - Misoprostol for induction of labour at term: a more effective agent than dinoprostone vaginal gel. AB - OBJECTIVE: To compare the efficacy of vaginal misoprostol and dinoprostone vaginal gel for induction of labour at term. DESIGN: A single-blind randomised comparative trial. SETTING: Induction and labour wards of a UK teaching hospital. PARTICIPANTS: Two hundred and eleven pregnant women at term in whom induction of labour was indicated, and with no contra-indication to the use of prostaglandins for the induction of labour. INTERVENTION: The women were randomly assigned to receive vaginal administration of either misoprostol 50 microg four hourly (to a maximum of four doses) or dinoprostone gel 1 mg six hourly (to a maximum of three doses). MAIN OUTCOME MEASURES: Time from induction to delivery, oxytocin requirement in labour, analgesic requirement, mode of delivery, neonatal outcome. RESULTS: The misoprostol group had a highly significant reduction in median induction-delivery interval compared with the dinoprostone group (14.4 hours vs 22.9 hours; P < 0.00001). In addition, more women delivered after only one dose (77% vs 49%; P < 0.0001, OR 3.51, 95% CI 1.94-6.35), and within 12 and 24 hours. There was a reduced need for oxytocin augmentation in labour (21% vs 47%; P < 0.0001, OR 0.30, 95% CI 0.16-0.54). There was no difference in analgesia requirement in labour, or in mode of delivery. There were no adverse neonatal outcomes associated with the use of misoprostol. Women in the misoprostol group experienced more pain in the interval between induction and being given analgesia in labour, but this did not reach statistical significance. CONCLUSIONS: Misoprostol 50 microg vaginally is a more effective induction agent than 1 mg dinoprostone vaginal gel, with no apparent adverse effects on mode of delivery, or on the fetus. The higher pain scores in the misoprostol group must be balanced against the reduction in time spent having labour induced, and the reduction in need for intravenous oxytocin augmentation. Further randomised studies must continue to exclude the possibility of rare adverse side effects. PMID- 10453829 TI - Misoprostol for induction of labour: a systematic review. AB - OBJECTIVE: To determine, from the best available evidence, the effectiveness and safety of misoprostol administered vaginally or orally for third trimester cervical ripening or induction of labour. METHODS: Clinical trials of misoprostol used for cervical ripening or labour induction in the third trimester were identified from the register of randomised trials maintained by the Cochrane Pregnancy and Childbirth Group. All identified trials were considered for inclusion in the review according to a prespecified protocol. Primary outcomes were chosen to address clinical effectiveness (delivery within 24 hours) and safety (uterine hyperstimulation, caesarean section, serious maternal and neonatal morbidity) and were determined a priori. All meta-analyses were based on the intention-to-treat principle. In the absence of heterogeneity the summary statistics have been expressed as typical relative risk (RR) and 95% confidence interval (CI). RESULTS: Vaginal misoprostol: one small study showed that the use of misoprostol results in more effective cervical ripening and reduced need for oxytocin when compared with placebo. When compared with oxytocin, vaginal misoprostol was more effective for labour induction. The relative risk of failure to achieve vaginal delivery within 24 hours was 0.48 (95% CI 0.35 to 0.66). However, the relative risks for uterine hyperstimulation with and without fetal heart rate abnormalities were 2.54 (95% CI 1.12 to 5.77) and 2.96 (95% CI 2.11 to 4.14), respectively. In three out of four trials which studied women with intact membranes and unfavourable cervices, failure to achieve vaginal delivery within 24 hours was reduced with misoprostol when compared with other prostaglandins (RR 0.71, 95% CI 0.62 to 0.81). Vaginal misoprostol was associated with increased uterine hyperstimulation both without fetal heart rate changes (RR 1.67, 95% CI 1.30 to 2.14) and with associated fetal heart rate changes (RR 1.45, 95% CI 1.04 to 2.04). There was also an increase in meconium stained amniotic fluid following vaginal misoprostol (RR 1.38, 95% CI 1.06 to 1.79). Oral misoprostol: one small trial suggests that, when compared with placebo, oral misoprostol reduces the need for oxytocin and shortens the time between induction and delivery. Compared with other prostaglandins one small trial showed a reduced need for oxytocin with oral misoprostol. Two trials compared oral with vaginal misoprostol using different doses. No significant differences were evident. CONCLUSIONS: Overall, misoprostol appears to be more effective than conventional methods of cervical ripening and labour induction. Although no differences in perinatal outcome were shown, the studies were not sufficiently large to exclude the possibility of uncommon serious adverse effects. In particular the increase in uterine hyperstimulation with fetal heart rate changes following misoprostol is a matter for concern. It is possible that, if sufficient numbers are studied, an unacceptably high number of serious adverse events including uterine rupture and asphyxial fetal deaths may occur. The data at present are not robust enough to address the issue of safety. Thus, though misoprostol shows promise as a highly effective, inexpensive and convenient agent for labour induction, it cannot be recommended for routine use at this stage. Lower dose misoprostol regimens should be investigated further. PMID- 10453830 TI - Randomised trial comparing expectant with medical management for first trimester miscarriages. AB - OBJECTIVE: To compare the efficacy of antiprogesterone (mifepristone) in combination with a synthetic prostaglandin E1 analogue (misoprostol) for outpatient treatment of miscarriages. PARTICIPANTS: One hundred and twenty-two women with first trimester miscarriages. METHODS: The women were randomised to treatment with mifepristone 400 mg orally followed by a single oral dose of 400 microg misoprostol 48 hours later (n = 60) or expectant management (n = 62). Women were re-evaluated five days later. If retained intrauterine products of conception were found with an antero-posterior diameter above 15 mm on transvaginal ultrasound, surgical evacuation was performed. RESULTS: Eighty-two percent of the women randomised to pharmacological treatment and 76% of those randomised to expectant management had an empty uterine cavity after five days. Convalescence time was 1.8 days longer for women randomised to pharmacological treatment. Pain, bleeding, complications, and satisfaction with the treatment did not differ between the groups. CONCLUSIONS: Most cases of spontaneous incomplete miscarriage will become a complete miscarriage without intervention. This study shows that outpatient treatment with a combination of antiprogesterone and a prostaglandin E1 analogue did not increase the rate of complete miscarriage, compared with expectancy alone, by a clinical important degree. PMID- 10453831 TI - Investigation of the cause of miscarriage and its influence on women's psychological distress. AB - OBJECTIVE: To determine whether knowledge of the possible cause of miscarriage reduces women's long term psychological distress. DESIGN: Prospective longitudinal study. SETTING: Harris Birthright Research Centre for Fetal Medicine, King's College Hospital Medical School, London. METHODS: In 143 women where a routine ultrasound scan at 10-14 weeks of gestation showed an anembryonic pregnancy or fetal death, investigations were carried out to help ascertain the cause of the pregnancy loss. The participants were divided into two groups according to whether the cause was identified or not, and the psychological conditions of the two groups were compared at four weeks and four months after the diagnosis of fetal loss. MAIN OUTCOME MEASURES: Anxiety, depression, grief, self-blame, worry. RESULTS: The scores for all outcome variables were significantly lower at the four-month compared with the four-week post miscarriage assessment. A fetal chromosomal abnormality was the most commonly identified cause of miscarriage, and this group reported significantly less self blame than women in whom no cause was identified. There were no significant differences between the groups on any other outcome variables. CONCLUSIONS: In women with a missed miscarriage, identification of the cause of fetal loss reduces the feelings of self-blame. PMID- 10453832 TI - Association of cerebral perfusion pressure with headache in women with pre eclampsia. AB - OBJECTIVE: To study estimated cerebral perfusion pressure and its relation to headache and scotomata in women with pre-eclampsia. DESIGN: Prospective, observational study. SETTING: University teaching hospitals. POPULATION: Seventy nine pre-eclamptic women with (n = 42) and without (n = 37) headache. Patients with scotomata were also studied separately. METHODS: Transcranial Doppler ultrasound was used to estimate the resistance index, pulsatility index, and estimated cerebral perfusion pressure in the middle cerebral artery. eCPP data were plotted on the same axes as the mean (and 5th and 95th% prediction limits) eCPP data from 63 normal pregnant women followed longitudinally through pregnancy. Data outside of the 95% prediction limits were regarded as abnormal. Data from the pre-eclamptic women were also expressed in terms of the number of standard deviations from the mean value established for normal pregnancy (multiples of the standard deviation: MOS). All studies were prior to labour, under similar conditions, and before volume expansion or treatment. Analysis of data was performed using Student's t test, Mann-Whitney U test, ANOVA, and Fisher's exact test with two-tailed P < 0.05, and receiver operating characteristic curve analysis with a one-tailed P < 0.05. MAIN OUTCOME MEASURES: Resistance index, pulsatility index, and eCPP. RESULTS: Pre-eclamptic women with headache were much more likely to have abnormal eCPP (34/42; 88%) than those without headache (18/37; 49%), P = 0.004, OR 4.5 (95% CI 1.5 to 13.9). There were no differences in terms of MOS in the resistance index or pulsatility index between the two groups, but estimated perfusion pressure, expressed as multiples of the standard deviation in the group with headache, was significantly higher than in the women without. Headache was noted in both over-perfusion and under perfusion states. Only women with headache had scotomata, and their presence was not related to the severity of the headache or any difference in resistance indices or eCPP. CONCLUSIONS: Headache in women with pre-eclampsia is strongly associated with the presence of abnormal cerebral perfusion pressure. This information may be of use in clinical management. PMID- 10453833 TI - Neutrophil function in women with pre-eclampsia. AB - OBJECTIVE: To investigate the function of neutrophils in normal pregnancy and pre eclampsia. DESIGN: Baseline levels and activated responses of peripheral blood neutrophils were measured in response to the physiological agonists, n-formyl-met leu-phe (fMLP) and zymosan activated serum. SAMPLE: Neutrophils of 16 pre eclamptic, 17 normal pregnant (third trimester) and 15 nonpregnant age-matched control women were calculated. SETTING: Antenatal Clinic, City Hospital, Nottingham. METHODS: Neutrophil superoxide anion production was determined by lucigenin-enhanced chemiluminescence; the release of secondary granule lactoferrin by ELISA; and the expression of cell surface adhesion molecules CD11b, CD18 and L-selectin (CD62L) by flow cytometric analysis. RESULTS: Superoxide anion generation was reduced in the pregnant group compared with nonpregnant controls [fMLP by 51% (P = 0.03) and zymosan activated serum by 56% (P = 0.01)] but pre-eclamptic measurements did not show a similar reduction. There were no differences between the three study groups in the plasma levels of lactoferrin, or in the stimulated expression and release of CD11b and CD18, or lactoferrin and L-selectin. The base-line levels for the production of superoxide anions; the expression of CD11b or CD18 or L-selectin; and the release of lactoferrin showed no significant differences. CONCLUSIONS: Circulating neutrophils in pregnancy and pre-eclampsia are neither activated nor primed in vivo, however the release of reactive oxygen species is diminished in normal pregnancy. In comparison, an elevation of reactive oxygen generation in pre eclampsia may highlight a role for neutrophils in the oxidative stress and pathophysiology of this disease. PMID- 10453834 TI - Early diagnosis of major cardiac defects in chromosomally normal fetuses with increased nuchal translucency. AB - OBJECTIVE: To determine the value of early ultrasound examination for prenatal diagnosis of cardiac defects in chromosomally normal fetuses with increased nuchal translucency thickness at 10-14 weeks of gestation. DESIGN: Prospective audit. SETTING: Fetal Medicine Centre. METHODS: Specialist fetal echocardiography was carried out in 398 chromosomally normal fetuses with a nuchal translucency measurement above the 99th centile (> or = 3.5 mm). In the first 75 pregnancies the cardiac scan was carried out at 17-22 weeks and the next 323 cases had one scan at 13-17 weeks and another at 20-22 weeks. Pregnancy outcome was obtained in all of the cases. RESULTS: Major cardiac defects were present in 29 (7.3% of 398) cases and in 28 of these the diagnosis was made by antenatal echocardiography. In 27 of the 29 fetuses with major cardiac defects echocardiography was carried out at 13-17 weeks and an abnormality was suspected in 24 of the cases (88%). CONCLUSIONS: Increased nuchal translucency in chromosomally normal fetuses is associated with an increased prevalence of major cardiac defects and, as such, is an indication for specialist fetal echocardiography. Most of the cardiac abnormalities are detectable on antenatal fetal echocardiography and many can be excluded by early scanning. PMID- 10453835 TI - Psychosocial predictors of low birthweight: a prospective study. AB - OBJECTIVE: To examine the role of psychosocial risk factors for low birthweight. DESIGN: A prospective study. SETTING: Obstetric outpatient clinics of the University Hospital Vrije Universiteit, Amsterdam. PARTICIPANTS: Three hundred and ninety-six nulliparous women. METHODS: Questionnaires on background variables, daily stressors, psychological and mental wellbeing, social support and work factors were completed by the women in the first, second and third trimester of pregnancy. Low birthweight for gestational age was defined at different cut off points: 1. < or = 10th customised birthweight centile (n = 69); 2. < or = 5th customised birthweight centile (n = 54); 3. < 3rd customised birthweight centile (n = 35); and 4. < or = the 10th Dutch birthweight centile (n = 40). Multivariate logistic regression was applied and the results were expressed in odds ratios and their 95% confidence intervals. RESULTS: When the cut off level was defined < or = 5th and < 3rd customised centile, the number of daily stressors in the first trimester was a statistically significant risk factor (OR 1.04, 95% CI 1.01-1.07 and OR 1.04, 95% CI 1.01-1.08). No significant psychosocial risk factors could be identified when low birthweight for gestational age was defined < or = the 10th customised birthweight centile. When low birthweight for gestational age was defined < or = the 10th Dutch birthweight centile, number of hours housekeeping per week in the first trimester (OR 1.59, 95% CI 1.03-2.46), low subjective severity rating of daily stressors in the first trimester (OR 0.41, 95% CI 0.17-0.97) and depressive mood in the first trimester (OR 1.12, 95% CI 1.01-1.24) were statistically significant psychosocial risk factors after controlling for maternal weight and height, number of cigarettes smoked per day and educational level. CONCLUSIONS: In the first trimester of pregnancy maternal psychosocial factors are associated with an increased risk of low birthweight. The specific psychosocial risk factors found were different when the definition of low birthweight was changed. Therefore, in this field of research, we suggest use of the most valid outcome measure for low birthweight, being the customised birthweight centiles. PMID- 10453836 TI - Urinary incontinence: prevalence and risk factors at 16 weeks of gestation. AB - OBJECTIVE: To evaluate the prevalence of urinary incontinence at 16 weeks of gestation and to identify possible maternal and obstetric risk factors. DESIGN: Cross-sectional study and cohort study. SETTING: Department of Obstetrics and Gynaecology, Aarhus University Hospital, Denmark. POPULATION: Cross-sectional study: 7795 women attending antenatal care. Cohort study: a sub-group of 1781 pregnant women with one previous delivery at our department. RESULTS: Prevalence and maternal risk factors: the prevalence of urinary incontinence within the preceding year was 8.9% among women at 16 weeks of gestation (nulliparae, 3.9%, para 1, 13.8%, para 2+, 16.2%). Stress or mixed incontinence occurred at least weekly in 3% of all the women. After adjusting for age, parity, body mass index, smoking, previous abortions, and previous lower abdominal or urological surgery in a logistic regression model, primiparous women who had delivered vaginally had higher risk of stress or mixed urinary incontinence than nulliparous women (OR 5.7; 95% CI 3.9-8.3). Subsequent vaginal deliveries did not increase the risk significantly. Young age, body mass index > 30, and smoking were possible risk factors for developing urinary incontinence. Obstetric factors: weight of the newborn > 4000 g (OR 1.9; 95% CI 1.0-3.6) increased the risk of urinary incontinence; mediolateral episiotomy in combination with birthweight > 4000 g also increased the risk (OR 3.5; 95% CI 1.2-10.2); a number of other intrapartum factors did not increase the risk of urinary incontinence. CONCLUSIONS: The first vaginal delivery was a major risk factor for developing urinary incontinence; subsequent vaginal deliveries did not increase the risk significantly. Birthweight > 4000 g increased the risk; episiotomy in combination with birthweight > 4000 g also increased the risk. PMID- 10453837 TI - Women's attitudes to urodynamics: a questionnaire survey. AB - OBJECTIVE: To obtain a measure of how well women tolerate urodynamic investigations and to determine how well they thought the test was explained. DESIGN: Prospective questionnaire survey. SETTING: A teaching hospital tertiary referral centre urodynamic laboratory. PARTICIPANTS: Three hundred and twenty four women attending for urodynamic investigations on 331 occasions. Questionnaires were returned from 297 women (91.7%) with six women returning two questionnaires. RESULTS: Urodynamic investigations were well tolerated by most women, with 45% feeling that the procedure was not as bad as they expected. Moderate or severe anxiety about the test was experienced by 42% and 40% felt moderately or severely embarrassed. Pain was noted by 27% of women during investigation, and by 13% after investigation. Overall distress from the procedure was less in older women and in those who had been referred from a specialist urogynaecology clinic. Distress was higher when difficulties were encountered during the investigation and in women who had investigations other than a standard cystometrogram. Women were likely to find the test less distressing when they felt they had been given adequate information about the test. CONCLUSIONS: Although urodynamic investigations are generally well tolerated, there is a significant minority of women who find the test embarrassing, painful and distressing. PMID- 10453838 TI - Use of oral contraceptives and uterine fibroids: results from a case-control study. AB - OBJECTIVE: To analyse the association between oral contraceptive use and the risk of uterine fibroids. DESIGN: We considered data collected in a case-control study on risk factors for uterine fibroids. PARTICIPANTS: We studied 843 women with uterine fibroids, whose clinical diagnosis dated back no more than two years. Controls were 1557 non-hysterectomised patients younger than 55 years admitted for acute, non-gynecological, non-hormonal, non-neoplastic conditions. RESULTS: A total of 254 cases (30.1%) and 360 controls (23.1%) reported ever using oral contraceptives: the odds ratio (OR) for ever vs never users was 1.1 (95% CI 0.8 1.3). The risk in current users was below unity when compared with never users (OR 0.3, 95% CI 0.2-0.6), while ex-users had a risk of fibroids comparable with never users (OR 1.1, 95% CI 0.9-1.4). The risk of uterine fibroids decreased with duration of oral contraceptive use: compared with never users, the estimated OR was 0.8 (95% CI 0.5-1.2) in ever users for four to six years and 0.5 (95% CI 0.3 0.9) for seven years or more (chi2 trend = 4.6, P = 0.03). CONCLUSIONS: Although the role of selection bias should be carefully evaluated, the present data suggest that uterine fibroids should not be considered a contra-indication for oral contraceptive use. PMID- 10453839 TI - Placental apoptosis is increased in post-term pregnancies. PMID- 10453840 TI - The role of high resolution ultrasound with guided cytology of groin lymph nodes in the management of squamous cell carcinoma of the vulva: a pilot study. AB - The accuracy of high resolution ultrasound with guided fine needle aspiration cytology in detecting inguinal lymph node involvement was assessed in 24 women undergoing radical vulvectomy and groin node dissection for squamous cell vulval cancer. Of the 43 groins dissected, ultrasound correctly diagnosed the lymph node status in 36, with five false positive and two false negative results. Cytology in 40 groins showed no false positive and five false negative results. The sensitivity and specificity for the combined techniques were 83% and 82% respectively. Assessed together, the combined technique failed to detect metastatic disease in two groins; in both cases the extent of nodal metastatic involvement was a solitary focus < 3 mm in diameter. The ultrasound and fine needle aspiration procedure is safe and well tolerated and can be repeated as needed for surveillance. The authors suggest that this procedure should be evaluated further to determine whether a policy of individual selection for lymphadenectomy can be implemented based on this technique. PMID- 10453841 TI - Sub-aponeurotic haemorrhage: a rare but life-threatening neonatal complication associated with ventouse delivery. PMID- 10453842 TI - Mitochondrial myopathy in a primigravid pregnancy. PMID- 10453843 TI - Insulin sensitivity in pre-eclampsia. PMID- 10453844 TI - Randomised trials in maternal and perinatal medicine: global partnerships are the way forward. PMID- 10453845 TI - Fetal ultrasound biometry: 1. Head reference values. PMID- 10453846 TI - The seven surgeons of King's: a fable by AEsop. PMID- 10453847 TI - A randomised trial of endometrial ablation versus hysterectomy for the treatment of dysfunctional uterine bleeding: outcome at four years. PMID- 10453848 TI - Steroids in acute exacerbation of asthma: how do we grade the evidence? PMID- 10453849 TI - Pericardiocentesis: blind no more! PMID- 10453850 TI - Office spirometry: does poor quality render it impractical? PMID- 10453851 TI - Tidal flow-volume analysis of ventilation during exercise: a useful approach for diagnosing the mechanism of ventilatory limitation to exercise during cardiopulmonary exercise testing. PMID- 10453852 TI - A pathogenic triad in chronic cough: asthma, postnasal drip syndrome, and gastroesophageal reflux disease. AB - BACKGROUND: Coughing may be produced by a number of different disorders in distinct anatomic sites. Chronic cough causes major functional limitation in a considerable patient population and requires careful evaluation. METHODS: Seventy eight nonsmoking patients of both genders who complained of cough for > or =3 weeks and had normal findings on plain chest radiographs were studied prospectively. Their histories were obtained, and physical examinations were performed. The diagnostic workup included pulmonary function tests, CT of the paranasal sinuses and chest, carbachol provocation test, fiberoptic rhinoscopy, fiberoptic bronchoscopy, and 24-h esophageal pH monitoring. The final diagnosis depended on clinical, radiologic, and laboratory findings; a successful response to therapy was required for confirmation. RESULTS: The causes of chronic cough were determined in all patients. Coughing was due to a single cause in 30 patients (38.5%) and multiple causes in 48 patients (61.5%). The five most important causative factors were asthma (46 patients; 58.9%), postnasal drip syndrome (PNDS; 45 patients; 57.6%), gastroesophageal reflux disease (GERD; 32 patients; 41.1%), bronchiectasis (14 patients; 17.9%), and tracheobronchial collapse (11 patients; 14.1%). INTERPRETATION: Asthma, PNDS, and GERD, alone or in combination, were responsible for 93.6% of the cases of chronic cough. The presence of these three conditions was so frequent that the expression "pathogenic triad of chronic cough" should be acknowledged in specialized literature. It is essential to consider pulmonary and extrapulmonary causes in order to prescribe a successful specific therapy for chronic cough. PMID- 10453853 TI - Corticosteroids in the emergency department therapy of acute adult asthma: an evidence-based evaluation. AB - OBJECTIVE: To review the literature to determine the benefits of corticosteroids (CCSs) (oral, IM, IV, or inhaled) in the treatment of adult patients with acute asthma presenting at an acute-care setting. SEARCH STRATEGY: A MEDLINE search was conducted using the following terms: (1) Asthma OR Wheez, AND (2) Glucocorticoids OR Steroids, AND (3) Acute OR Emerg. Other sources were the CURRENT CONTENTS database, review articles, reference sections of located studies, and a manual search of the top 15 journals for respiratory and emergency medicine. SELECTION CRITERIA: Patients were selected for the study by the following criteria: (1) English language; (2) adult patients with asthma whose acute exacerbations were the primary reason for assessment; (3) involvement in randomized, controlled trials conducted in an emergency care setting; (4) patients had participated in a study investigating a primary research question involving treatment with CCSs; and (5) outcomes based on results of pulmonary function tests and on hospital admission rates. RESULTS: At the 3-h assessment, only high doses of inhaled CCSs significantly improved pulmonary function compared with placebo (effect size [ES], 0.56; 95% confidence interval [CI], 0.15 to 0.97). On the other hand, after receiving IV CCSs, patients required at least 6 to 24 h to show moderate but nonsignificant improvements of pulmonary function (6-h ES, 0.44 [95% CI, -0.01 to 0.89]; 12-h ES, 0.54 [95% CI, -0.08 to 1.17]; and 24-h ES, 0.53 [95% CI, -0.39 to 1.45]). The data from the six studies that we used to pool information on admission rate outcome showed a 32% reduction in favor of the use of IV CCSs (relative risk [RR], 0.68 [95% CI, 0.47 to 0.99]; number needed to treat, 12.5 [95% CI, 7.1 to 50]). However, the pooled effect of the three high-quality studies showed no difference between groups (RR, 1.21; 95% CI, 0.67 to 2.18). Oral CCSs provided a similarly beneficial effect on pulmonary function when compared with parenteral administration (ES, -0.14; 95% CI, -0.82 to 0.31. Finally, the results showed a nonsignificant favorable trend toward improved outcome with medium or high doses of CCSs. CONCLUSIONS: This evidence-based evaluation suggests that the administration of parenteral CCSs to the patient on arrival at the emergency department (ED) neither improves airflow obstruction nor reduces the need for hospitalization. Parenteral CCSs probably require >6 to 24 h to begin to act. Comprehensible conclusions about admission rates in the ED setting are difficult to make. At the 3-h assessment, only high doses of inhaled CCSs (in one study) significantly improved pulmonary function compared with placebo. IV and oral CCSs appear to have equivalent effects, and there is a tendency toward improvement in pulmonary function with medium or high doses. PMID- 10453854 TI - The effect of heliox in acute severe asthma: a randomized controlled trial. AB - STUDY OBJECTIVES: To evaluate the effect of heliox on airflow obstruction and dyspnea in patients with acute severe asthma. DESIGN: A prospective, randomized, controlled study. SETTING: A university hospital. PATIENTS: Twenty-three patients presenting to the emergency department with acute severe asthma were randomized to receive 70%/30% heliox or 30% oxygen. MEASUREMENTS: Peak expiratory flow (PEF), dyspnea score, heart rate, respiratory rate (RR), and BP were measured at baseline and 20, 120, 240, 360, and 480 min after starting the test gas. After baseline, the PEF was measured by using the gas that was randomized to the treatment program. RESULTS: In the first 20 min, there was a 58.4% increase in percent predicted PEF (%PEF) in the heliox group (p<0.001), whereas there was only a 10.1% increase in %PEF for the oxygen group (p>0.1). Eighty-two percent of the heliox group had >25% improvement in %PEF at 20 min, whereas only 17% of the oxygen group did (p<0.01). The next significant improvement in %PEF in the heliox group occurred at 480 min. At the end of the study in the heliox group, the PEF did not significantly (p>0.1) change immediately after the heliox was discontinued (270.6 to 264.2 L/min). In the heliox group in the first 20 min, there was a significant decrease in dyspnea score and RR (p<0.05), but there were no further significant improvements for the rest of the study. In the oxygen group, no variables significantly improved until 360 min. CONCLUSION: Heliox rapidly improves airflow obstruction and dyspnea in patients with acute severe asthma and may be useful as a therapeutic bridge until the corticosteroid effect occurs. PMID- 10453855 TI - Serum eosinophil cationic protein and bronchial responsiveness in pediatric and adolescent asthma patients. AB - BACKGROUND: Serum eosinophil cationic protein (ECP) has been promoted as a marker of inflammatory activity in bronchial asthma. Bronchial responsiveness, measured either by inhaling pharmacologically active substances such as histamine or methacholine, or by applying physical stimuli such as the hyperventilation of cold dry air, is also considered to be an indirect marker of bronchial inflammation. OBJECTIVES: In this study, we investigated the possible relationship between serum ECP and bronchial responsiveness to both cold dry air and histamine in presently symptom- and medication-free pediatric and adolescent asthma patients. SUBJECTS: Thirty-six children and adolescents with atopic asthma were studied. METHODS: On 2 consecutive days, bronchial responsiveness was assessed nonpharmacologically by cold dry air and pharmacologically by histamine in random order. Blood samples for determination of ECP were collected before each challenge. RESULTS: Serum ECP levels correlated with neither cold dry air induced changes in FEV1 nor the provocation concentrations of histamine causing a 20% fall in FEV1. Subjects with bronchial hyperresponsiveness to cold dry air and histamine had somewhat higher levels of serum ECP than subjects with normal responses, but these differences were insignificant. CONCLUSIONS: Our results indicate a lack of relationship both between serum ECP and bronchial responsiveness to cold dry air and between serum ECP and bronchial responsiveness to histamine. PMID- 10453856 TI - Trends in the epidemiology of COPD in Canada, 1980 to 1995. COPD and Rehabilitation Committee of the Canadian Thoracic Society. AB - PURPOSE: To describe trends in the epidemiology of COPD in Canada from 1980 to 1995, in terms of perceived prevalence, mortality, and hospital morbidity. DATA SOURCES: We limited the analysis to data related to chronic bronchitis, emphysema, or chronic airway obstruction not classified elsewhere, and excluded asthma (Ninth International Classification of Diseases, codes 490 to 492 and 496). The perceived prevalence rate of COPD was derived from the 1994-1995 National Health Survey. Mortality and hospital morbidity data (from 1980 to 1995) were obtained from the Health Statistics Division of Statistics Canada. RESULTS: From the National Health Survey, it was estimated that 750,000 Canadians had chronic bronchitis or emphysema diagnosed by a health professional. Prevalence rates were the following: ages 55 to 64 years, 4.6%; ages 65 to 74 years, 5.0%; > or =75 years, 6.8%. From 1980 to 1995, the total number of deaths from COPD increased from 4,438 to 8,583. Although the age-standardized mortality rate remained stable throughout this period in men (around 45/100,000 population), it doubled in women (8.3/100,000 in 1980 to 17.3/100,000 in 1995). There were 55,782 hospital separations in 1993-1994 with COPD as the primary discharge diagnosis (compared to 42,102 in 1981-1982). In people aged > or =65 years, the age specific hospital separation rate increased over this period, especially in women > or =75 years (from 504/100,000 to 1,033/100,000). The average in-hospital length of stay was 9.6 days in 1981-1982 and 8.3 days in 1993-1994. CONCLUSION: COPD represents a major health issue in Canada and will likely remain so for decades. Physician and non-physician health professionals who provide health care, as well as those who fund it must actively encourage approaches for primary and secondary prevention of this condition as well as approaches shown to be effective in addressing its associated impairment, disability, and handicap. PMID- 10453857 TI - Ambulatory oximetry monitoring in patients with severe COPD: a preliminary study. AB - BACKGROUND: The benefits of long-term oxygen supplementation in COPD patients with hypoxemia are well established. The standard approach to prescribing oxygen uses a static assessment of oxygen requirements in a hospital or clinic setting. The assumption behind this approach is that patients will maintain a "therapeutic" hemoglobin oxygen saturation (SpO2) in the outpatient setting. We questioned the validity of this assumption, and hypothesized that many patients may demonstrate significant oxygen desaturation during normal activities of daily living. STUDY DESIGN, METHODS, AND MEASUREMENTS: We determined if oxygen supplementation maintained a therapeutic SpO2 level in patients with COPD (n = 27), using the technique of ambulatory oximetry monitoring (AOM). AOM consisted of using a portable oximeter to monitor SpO2, pulse rate, and patient activity while patients were engaged in normal activities of daily living over an extended time period (approximately 18 h). The portable oximeter collected and stored these data every 15 s over the monitored time period. Each AOM recording was manually scored for desaturation events and other key variables, including average SpO2 over the monitoring period, the average number of desaturation events per hour, and the percentage of monitored time deleted secondary to artifacts. SETTING: University-affiliated Veterans Affairs Medical Center. PATIENTS: All subjects were patients with stable COPD with no recent history of hospitalization or exacerbation of their lung disease. RESULTS: This cohort of patients demonstrated a surprising frequency of desaturation below the recommended target SpO2 value (90%), which averaged approximately 25% of AOM recording time. There was wide variability among patients in the percentage of time SpO2 was below the target value (range, 3 to 67% of AOM recording time). Motion artifact on the AOM recordings was not a major problem; an average of 8% of the recording time was deleted secondary to artifacts in this patient cohort. CONCLUSIONS: The results demonstrate that AOM is feasible and accurate with an acceptable level of motion artifact. These results also suggest that the standard approach for prescribing oxygen may lead to subtherapeutic SpO2 values in the outpatient setting. AOM holds promise as a tool to monitor the adequacy of oxygen prescriptions in the outpatient setting in patients with lung disease. PMID- 10453858 TI - Clinical and echocardiographic characteristics of significant pericardial effusions following cardiothoracic surgery and outcomes of echo-guided pericardiocentesis for management: Mayo Clinic experience, 1979-1998. AB - STUDY OBJECTIVES: This study assessed the clinical features, timing of presentation, and echocardiographic characteristics associated with clinically significant pericardial effusions after cardiothoracic surgery. The outcomes of echocardiographically (echo-) guided pericardiocentesis for the management of these effusions were evaluated. DESIGN: From the prospective Mayo Clinic Registry of Echo-guided Pericardiocentesis (February 1979 to June 1998), 245 procedures performed for clinically significant postoperative effusions were identified. Clinical features, effusion causes, echocardiographic findings, and management outcomes were studied and analyzed. Cross-referencing the registry with the Mayo Clinic surgical database provided an estimate of the incidence of significant postoperative effusions and the number of cases in which primary surgical management was chosen instead of pericardiocentesis. RESULTS: Use of anticoagulant therapy was considered a significant contributing factor in 86% and 65% of early effusions (< or =7 days after surgery) and late effusions (>7 days after surgery), respectively. Postpericardiotomy syndrome was an important factor in the development of late effusions (34%). Common presenting symptoms included malaise (90%), dyspnea (65%), and chest pain (33%). Tachycardia, fever, elevated jugular venous pressure, hypotension, and pulsus paradoxus were found in 53%, 40%, 39%, 27%, and 17% of cases, respectively. Transthoracic echocardiography permitted rapid diagnosis and hemodynamic assessment of all effusions except for three cases that required transesophageal echocardiography for confirmation. Echo guided pericardiocentesis was successful in 97% of all cases and in 96% of all loculated effusions. Major complications (2%), including chamber lacerations (n = 2) and pneumothoraces (n = 3), were successfully treated by surgical repair and chest tube reexpansion, respectively. Median follow-up duration for the study population was 3.8 years (range, 190 days to 16.4 years). The use of extended catheter drainage was associated with reduction in recurrence for early and late postoperative effusions by 46% and 50%, respectively. CONCLUSIONS: The symptoms and physical findings of clinically significant postoperative pericardial effusions are frequently nonspecific and may be inadequate for a decision regarding intervention. Echocardiography can quickly confirm the presence of an effusion, and pericardiocentesis under echocardiographic guidance is safe and effective. The use of a pericardial catheter for extended drainage is associated with lower recurrence rates, and the majority of patients so treated do not require further intervention. PMID- 10453859 TI - Lung function and cardiopulmonary exercise performance after heart transplantation: influence of cardiac allograft vasculopathy. AB - STUDY OBJECTIVE: The reduced exercise capacity observed in most patients after heart transplantation may be due to treatment with immunosuppressive drugs, deconditioning, cardiac denervation, and graft rejection. Cardiac allograft vasculopathy (CAV) is presently the major factor limiting long-term survival after transplantation. Little information is available with regard to the relationship between CAV and functional impairment in these patients. DESIGN: Prospective. SETTING: A university hospital and a large transplant center. PATIENTS: About 37+/-5 months (range, 2 to 137 months) after orthotopic heart transplantation, 120 patients underwent lung function testing, cardiopulmonary exercise testing, and right and left heart catheterization. Significant CAV was defined as a stenosis > or =70% or severe diffuse obliteration in any of the three main vessels. Group I (n = 28) had a significant CAV; group II (n = 92), without a remarkable CAV, was the control group. MEASUREMENTS AND RESULTS: Overall, the maximum heart rate was 86+/-2% of what was predicted, and the peak oxygen consumption was 18.8+/-0.7 mL/kg/min (64% of that predicted). Groups I and II did not show significant differences with regard to anthropometric data, hemodynamic measurements, or number of rejection episodes. Group I exhibited significant differences in maximum heart rate (120+/-5 vs. 134+/-3 beats/min; p<0.01), work capacity (47+/-5% vs. 59+/-3%; p<0.05), peak oxygen uptake (16+/-1 vs. 20+/-1 mL/min/kg; p<0.01), and functional dead space ventilation (31+/-2 vs. 26+/-1; p<0.01). Pretransplant status, etiology of heart failure, ischemic time, and the number of rejection episodes did not correlate with any exercise parameter. CONCLUSIONS: Following heart transplantation, patients with significant CAV show a diminished exercise capacity, a reduced oxygen uptake, and a ventilation-perfusion mismatch. Thus, CAV may be a major factor limiting exercise capacity in heart-transplant patients. PMID- 10453860 TI - Detection and evaluation of asymptomatic myocarditis in schoolchildren: report of four cases. AB - STUDY OBJECTIVE: Data on the prevalence of myocarditis in children are limited. Autopsy studies have shown that myocarditis is often undiagnosed. We attempted to investigate the clinical features of asymptomatic myocarditis in four schoolchildren detected during a mass ECG screening in schoolchildren. DESIGN AND SETTING: To evaluate asymptomatic myocarditis, we clinically examined 12 schoolchildren who were referred to Shiga University of Medical Science Hospital, Otsu, Japan, because of abnormal ST or T waves detected during ECG screening. None of the 12 children had experienced any episodes suggesting cardiac disease or Kawasaki disease. Cardiac function and myocardial viability were assessed by two-dimensional echocardiography (2-DE), thallium-201 (201Tl) myocardial scintigraphy, and cardiac catheterization. Endomyocardial biopsy specimens were examined histologically. PATIENTS: Endomyocardial biopsy specimens revealed histologic evidence of myocarditis in 4 of the 12 children with abnormal ST or T waves. RESULTS: Abnormal tracer perfusion was observed on 201Tl myocardial scintigrams in these four children, but the results of coronary arteriography were normal. 2-DE showed left ventricular hypokinesis in one child and left ventricular enlargement in one of the four children with histologic evidence of myocarditis. A second endomyocardial biopsy specimen was obtained in two of four children, showing persistent myocarditis in one child. CONCLUSIONS: This type of screening program and indepth evaluation using 2-DE and 201Tl myocardial scintigraphy appear to be helpful in identifying children with myocarditis. The present histologic investigations suggested that even asymptomatic myocarditis might result in persistent heart damage. PMID- 10453861 TI - Cardiology or primary care for heart failure in the community setting: process of care and clinical outcomes. AB - STUDY OBJECTIVES: Severity of illness, treatment choices, and clinical outcomes may vary with physician training. This study was performed to determine whether such differences exist among patients with congestive heart failure (CHF) treated by cardiologists and by noncardiologists in the community hospital setting. DESIGN: Prospective cohort study. SETTING: Ten acute-care community hospitals. PATIENTS, MEASUREMENTS, AND RESULTS: Two thousand four hundred fifty-four patients with CHF were identified and followed up for 6 months after hospital discharge. Patients who were not treated by a cardiologist (group I; n = 977) were compared with patients whose attending physician was a cardiologist (group II; n = 419) and patients who received consultative care from a cardiologist (group III; n = 1,058). When compared with group I patients, group II patients were more likely to receive the recommended diagnostic tests and treatment strategies, although some of these differences could be explained by variations in the case mix. Group II patients had higher hospital charges, but lower CHF readmission rates and better postdischarge quality-of-life measures. No differences in adjusted mortality rates were observed. CONCLUSIONS: In the community-hospital setting, the clinical practices of cardiologists are more compatible with published treatment guidelines than the clinical practices of other physicians. The benefits of cardiology specialty care include lower CHF readmission rates and better postdischarge quality-of-life measures, rather than lower mortality rates, fewer hospital charges, or shorter length of stay. PMID- 10453862 TI - Cardiopulmonary exercise testing as a screening test for perioperative management of major surgery in the elderly. AB - STUDY OBJECTIVE: To develop an integrated strategy for the identification and subsequent management of high-risk patients in order to reduce both morbidity and mortality. DESIGN: Prospective consecutive series in which all patients underwent cardiopulmonary exercise (CPX) testing. SETTING: CPX laboratory and level 3 ICU and high-dependency unit (HDU) of a metropolitan teaching hospital. PATIENTS: Five hundred forty-eight patients >60 years of age (or younger with known cardiopulmonary disease) scheduled for major intra-abdominal surgery. INTERVENTIONS: The patients were assigned to one of three management strategies (ICU, HDU, or ward) based on the anaerobic threshold (deltaT) and ECG evidence of myocardial ischemia as determined by CPX testing that was performed as part of the presurgery evaluation, and by the expected oxygen demand stress of the surgical procedure. RESULTS: Overall mortality was 3.9%. Forty-three percent of deaths were attributed to poor cardiopulmonary function, as detected preoperatively. There were no deaths related to cardiopulmonary complications in any patient deemed fit for major abdominal surgery and ward management, as determined by CPX testing. CONCLUSIONS: In elderly patients undergoing major intra-abdominal surgery, the AT, as determined by CPX testing, is an excellent predictor of mortality from cardiopulmonary causes in the postoperative period. Preoperative screening using CPX testing allowed the identification of high-risk patients and the appropriate selection of perioperative management. PMID- 10453863 TI - Reproducibility of maximal exercise ergometer testing in patients with cystic fibrosis. AB - OBJECTIVES: Exercise testing in patients with cystic fibrosis (CF) has become an important tool in assessing disease severity and predicting overall outcome. The reproducibility of maximal exercise testing was examined in adult subjects with stable CF. METHODS: Nine subjects with CF underwent a total of three maximal exercise tests carried out under identical circumstances over a 28-day period. Oxygen uptake (VO2), minute ventilation (VE), respiratory frequency (f), heart rate (HR), and arterial oxygen saturation (SaO2) were measured at rest, at end exercise, and at 40% and 70% of maximum workload. RESULTS: There were no significant differences in these measurements among the three tests. Reproducibility of exercise performance was assessed using the coefficient of variation. The mean within-subject coefficient of variation for test variables at end exercise are as follows: VO2, 6.9%; VE, 6.2%; f, 5.8%; IIR, 3.0%; and SaO2, 1.1%. The mean within-subject coefficient of variation for test variables at 40% and 70% of maximal work rates are as follows: VO2, 5.2% and 4.6%; SaO2, 0.3% and 0.9%; HR, 4.0% and 3%; VE, 5.7% and 6.5%; and f, 5.8% and 7.2%, respectively. CONCLUSIONS: Variables measured during clinical cycle ergometer exercise testing in adult patients with stable CF are reproducible. No learning effect was found on repeated testing. PMID- 10453864 TI - The penetration of ceftibuten into the respiratory tract. AB - STUDY OBJECTIVE: To determine the penetration of ceftibuten into various respiratory tissues and fluids. DESIGN: Single-dose, open-label, pharmacokinetic study. SETTING: Veterans Administration Medical Center. PATIENTS: Twelve hospitalized men aged 34 to 75 years with a variety of noninfectious pulmonary symptoms/diseases. INTERVENTIONS: Patients received a single oral dose of ceftibuten, 200 mg, prior to undergoing diagnostic fiberoptic bronchoscopy. Plasma samples for the determination of ceftibuten concentrations were collected pretreatment and up to 12 h postdosing. Nasal secretions, tracheal secretions, BAL fluid, and lung tissue from a biopsy were obtained at bronchoscopy from 2 to 7 h postdosing. MEASUREMENTS AND RESULTS: Mean pharmacokinetic parameters for ceftibuten in plasma were the following: maximum observed plasma concentration (Cmax), 8.77 microg/mL; time to reach Cmax, 2.2 h; area under the plasma concentration-time curve extraploated to infinity, 49.21 microg/h/mL; and terminal elimination half-life, 3.17 h. These parameters were similar to those obtained in studies using healthy volunteers. Mean penetration of ceftibuten into nasal, tracheal, and bronchial secretions was 47%, 50%, and 30%, respectively. Mean penetration into BAL fluid was 81%, whereas penetration into lung tissue was 39%. No patient experienced any adverse effects related to ceftibuten. CONCLUSIONS: Ceftibuten penetrates well into various tissues and fluids of the upper and lower respiratory tracts. The results support the activity of ceftibuten in the treatment of upper and lower respiratory tract infections. PMID- 10453865 TI - Proportion of community-acquired pneumonia cases attributable to tobacco smoking. AB - STUDY OBJECTIVES: To assess the population-attributable risk (PAR) of smoking and the effect of tobacco smoking on the development of community-acquired pneumonia (CAP) in adults. DESIGN: A population-based case-control study. Information on history of smoking and other risk factors was obtained by a questionnaire administered by interview. SETTING: Mixed residential-industrial area having 74,610 adult inhabitants in Barcelona, Spain. PARTICIPANTS: Two hundred five male and female patients (age, 15 to 74 years old) with CAP diagnosed between 1993 and 1995. They were matched to 475 control subjects randomly selected from the municipal census. RESULTS: Smoking any type of tobacco had an odds ratio (OR) of CAP of 2.0 for ever smokers (95% confidence interval [CI], 1.24 to 3.24); 1.88 for current smokers (95% CI, 1.11 to 3.19); and 2.14 for ex-smokers (95% CI, 1.26 to 3.65). A positive trend for increased risk of CAP was observed for an increase in the duration of the habit, the average number of cigarettes smoked daily, and cumulative cigarette consumption. Former smokers had a 50% reduction in the OR 5 years after the cessation of smoking. The risk of CAP attributable to the consumption of any type of tobacco in this population was 32.4% of cases (95% CI, 14.8 to 50.1%). In subjects without a history of COPD, the PAR of tobacco was 23.0% (95% CI, 3.3 to 42.7%). CONCLUSION: This study gives better quantitative and qualitative evidence about the effects of tobacco smoking on the occurrence of pneumonia in the adult community. PMID- 10453866 TI - The clinical host response to microbial infection in medical patients with fever. AB - STUDY OBJECTIVES: Predictors among demographic, clinical, and laboratory variables for a microbial (nonviral/nonchlamydial) infection in hospitalized medical patients with new onset of fever (temperature > or =38.0 degrees C axillary or > or =38.3 degrees C rectal) were analyzed and compared with the criteria for the systemic inflammatory response syndrome (SIRS), including an abnormal body temperature and WBC count, tachypnea and tachycardia, and sepsis, defined as SIRS and the presence of a clinical infection. DESIGN: A prospective cohort study. SETTING: Department of internal medicine at a university hospital. PATIENTS: In 300 hospitalized medical patients with new onset of fever, demographic, clinical, and laboratory variables were obtained during the first 2 days after inclusion, and peak and nadir values, when appropriate, were taken. Microbiologic results for 7 days were collected. Clinical information was used to decide on the presence of a clinical infection. MEASUREMENTS AND RESULTS: One hundred thirty-three of 300 patients (44%) had a microbial infection: 26% suffered from local microbial infection only, 9% from bacteremia only, and 9% had bloodstream plus local microbial infections. Patients with a microbial infection had a higher World Health Organization performance score at home (p<0.05), higher peak body temperature (p<0.001), higher nadir and peak WBC counts (p<0.05), lower nadir platelet count (p<0.01), higher peak alanine and aspartate aminotransferases (p<0.01), and lower nadir albumin (p<0.001) levels in blood during the first 2 days after inclusion than those without infection. Using multivariate techniques, predictors for microbial infection or bacteremia alone, independent of age, sex, underlying disease, and clinical infection, were peak temperature, peak WBC count, and nadir platelet count and albumin level. In contrast, conventional SIRS/sepsis definitions and criteria predicted microbial infection less well, mainly because tachypnea and tachycardia were of no predictive value. CONCLUSIONS: In febrile medical patients, microbial infection can be predicted with use of easily obtained clinical and laboratory variables, including peak temperature, peak WBC count, and nadir platelet count and albumin level within the first 2 days. The new model predicted microbial infection better than conventional SIRS/sepsis criteria. This may help to improve the clinical recognition of the systemic host response to microbial infection and to refine SIRS/sepsis definitions. PMID- 10453867 TI - Cisplatin, mitomycin, and interferon-alpha2a combination chemoimmunotherapy in the treatment of diffuse malignant pleural mesothelioma. AB - STUDY OBJECTIVE: To investigate the therapeutic activity and toxicity of combination chemoimmunotherapy with cisplatin, mitomycin, and interferon (IFN) alpha2a, by comparing the responses in a group of patients with diffuse malignant pleural mesothelioma (DMPM) to the responses in a control group of DMPM patients given supportive care alone. DESIGN: Patients with histopathologically confirmed DMPM were evaluated for treatment with chemoimmunotherapy. SETTING: After the initial evaluation, all patients received either chemoimmunotherapy or supportive care from the Osmangazi University Medical Faculty, Department of Chest Diseases. PATIENTS: Forty-three patients with DMPM received chemoimmunotherapy until the end of the survey; 19 patients were given supportive therapy alone after refusing chemoimmunotherapy. INTERVENTIONS: Drugs were administered according to the following schedule: IV cisplatin, 30 mg/m2 qd on days 1 and 2; IV mitomycin, 8 mg/m2 on day 1; and subcutaneous IFN-alpha2a, 4.5 million IU twice weekly. The courses were repeated every 4 weeks. RESULTS: Overall, 232 chemoimmunotherapy cycles were administered. A total of 10 objective responses (ORs) in 43 patients (23%) were assessed, including 2 complete responses (5%), 4 partial responses, and 4 regressions. Seventeen patients had stable disease, and 16 patients had progression. The median survival time was 11.5 months for the 43 patients who received chemoimmunotherapy and 7.0 months for the 19 patients who received supportive therapy alone. The difference in survival times between the chemoimmunotherapy and supportive therapy groups was not significant. However, the median survival time for the patients who had OR was 21.3 months, which is significantly longer than that of the patients who received supportive care alone and that of patients with progressive disease (6 months). The toxicities associated with the treatment schedule of this study were, for the most part, tolerable. CONCLUSIONS: The drug combination used in this study is moderately effective and well tolerated in patients with DMPM, especially in those who responded to the treatment. PMID- 10453868 TI - Modification of pleural fluid pH by local anesthesia. AB - BACKGROUND: It is a common practice to anesthetize patients before performing a thoracentesis. We postulated that this technique may cause a clinically significant difference in the pH of the pleural fluid. METHODS: We compared two methods of determining pleural fluid pH. Fifty patients undergoing diagnostic or therapeutic thoracentesis were enrolled. Two 4-mL aliquots of pleural fluid were anaerobically collected into blood gas syringes containing heparin, one before (group A) and the other after (group B) anesthetizing the patient with 5 mL of 2% mepivacaine. pH was then determined on both samples using an arterial blood gas machine. Agreement analysis was performed overall and in subcategories of pH used to define complicated (<7.1), borderline (7.1 to 7.3), or uncomplicated (>7.3) parapneumonic effusions. We analyzed these same data stratified by the volume of pleural fluid in relationship to the size of the hemithorax (<15% and >15%). RESULTS: There was a statistical difference between the mean pH in both groups (group A, 7.32; group B, 7.28; p<0.0001). There was a significant correlation between the two measures (r = 0.97; p<0.0001). Using the pH subcategories, there was 45% discordance in classification for patients with parapneumonic effusions. The pH values obtained in group B wrongly predicted whether the patient required a chest tube in two of four cases (50%). In patients with effusions that occupied <15% of the affected hemithorax, there was an 80% discordance in classification for patients with parapneumonic effusions, and the pH values obtained in group B wrongly predicted whether the patient required a chest tube in two of two cases (100%). CONCLUSIONS: Local anesthesia is typically used before thoracentesis is performed. However, in cases of suspected parapneumonic effusions that occupy <15% of the affected hemithorax, pH results may be significantly altered by use of local mepivacaine anesthesia. PMID- 10453869 TI - Effects of humidification on nasal symptoms and compliance in sleep apnea patients using continuous positive airway pressure. AB - STUDY OBJECTIVES: To evaluate the effects of humidification on nasal symptoms and compliance in sleep apnea patients using continuous positive airway pressure (CPAP). DESIGN: A randomized, crossover design was employed. SETTING: The study was conducted at two suburban community-based hospital sleep laboratories. PATIENTS: Data were collected on 38 obstructive sleep apnea patients (mean age, 44.1 years) in whom CPAP was a novel treatment. INTERVENTIONS: The interventions were heated humidity, cold passover humidity, and a washout period without humidity. MEASUREMENTS AND RESULTS: Patients were titrated with heated humidity or cold passover humidity in the laboratory and subsequently initiated on humidity. Objective compliance, self-report of factors affecting CPAP use, satisfaction with CPAP, feeling upon awakening, and daytime sleepiness were assessed at the completion of each 3-week treatment period and a 2-week washout period. Outcome measures were assessed with one-way analysis of variance followed by Scheffe post hoc comparisons. Significant main effects were observed for compliance (F2,37 = 5.2; p = 0.008), satisfaction with CPAP (F2,37 = 4.5; p = 0.01), and feeling refreshed on awakening (F2,37 = 4.4; p = 0.02). A significant decrease in daytime sleepiness was observed between baseline and each of the conditions (F3,37 = 55.5; p<0.0001), but Epworth sleepiness scale scores did not differ between conditions (all p values >0.56). CPAP use with heated humidity (5.52+/-2.1 h/night) was greater than CPAP use without humidity (4.93+/-2.2 h/night; p = 0.008). Compliance differences were not observed between CPAP use with cold passover humidity and CPAP use without humidity. Patients were more satisfied with CPAP when it was used with heated or cold passover humidity (p< or =0.05). However, only heated humidity resulted in feeling more refreshed on awakening (p<0.05). No significant differences were observed among the three groups on the global adverse side effect score (F2,37 = 2.5; p = 0.09). Specific side effects such as dry mouth or throat and dry nose were reported less frequently when CPAP was used with heated humidity compared to CPAP use without humidity (p<0.001). CONCLUSIONS: Compliance with CPAP is enhanced when heated humidification is employed. This is likely due to a reduction in side effects associated with upper airway symptoms and a more refreshed feeling upon awakening. Compliance gains may be realized sooner if patients are started with heated humidity at CPAP initiation. PMID- 10453870 TI - Neural network prediction of obstructive sleep apnea from clinical criteria. AB - STUDY OBJECTIVES: Clinical prediction models for the diagnosis of obstructive sleep apnea (OSA) have lacked the accuracy necessary to confidently replace polysomnography (PSG). Artificial neural networks are computer programs that can be trained to predict outcomes based on experience. This study was conducted to test the hypothesis that a generalized regression neural network (GRNN) could accurately classify patients with OSA from clinical data. STUDY DESIGN: Retrospective review. SETTING: Regional sleep referral center. PATIENTS: Randomly selected records of patients referred for possible OSA. MEASUREMENTS: The neural network was trained using 23 clinical variables from 255 patients, and the predictive performance was evaluated using 150 other patients. RESULTS: The prevalence of OSA in this series of 405 patients (293 men and 112 women) was 69%. The trained GRNN had an accuracy of 91.3% (95% confidence interval [CI], 86.8 to 95.8). The sensitivity was 98.9% for having OSA (95% CI, 96.7 to 100), and the specificity was 80% (95% CI, 70 to 90). The positive predictive value that the patient would have OSA was 88.1% (95% CI, 81.8 to 94.4), whereas the negative predictive value that the patient would not have OSA (if so classified) was 98% (95% CI, 94 to 100). CONCLUSIONS: Appropriately trained GRNN has the ability to accurately rule in OSA from clinical data, and GRNN did not misclassify patients with moderate to severe OSA. In this study, use of the neural network could have reduced the number of PSG studies performed. Prospective validation of the neural network for the diagnosis of OSA is now required. PMID- 10453871 TI - Spirometry in primary care practice: the importance of quality assurance and the impact of spirometry workshops. AB - OBJECTIVE: To determine the quality of spirometry performed in primary care practice and to assess the impact of formal training. DESIGN: Randomized, controlled prospective interventional study. SETTING: Primary care practice, Auckland City, New Zealand. PARTICIPANTS: Thirty randomly selected primary care practices randomized to "trained" or "usual" groups. One doctor and one practice nurse were nominated to participate from each practice. INTERVENTIONS: "Trained" was defined as participation in an "initial" spirometry workshop at week 0 and a "maintenance of standards" workshop at week 12. "Usual" was defined as no formal training until week 12, when participants they attended the same "initial" workshop provided for the trained group. The study duration was 16 weeks. Each practice was provided with a spirometer to be used at their clinical discretion. MEASUREMENTS AND RESULTS: Spirometry data were uploaded weekly and analyzed using American Thoracic Society (ATS) criteria for acceptability and reproducibility. The workshops were assessed objectively with practical and written assessments, confirming a significant training effect. However, analysis of spirometry performed in clinical practice by the trained practitioners revealed three acceptable blows in only 18.9% of patient tests. In comparison, 5.1% of patient tests performed by the usual practitioners had three acceptable blows (p<0.0001). Only 13.5% of patient tests in the trained group and 3.4% in the usual group (p<0.0001) satisfied full acceptability and reproducibility criteria. However, 33.1% and 12.5% of patient tests in the trained and usual groups, respectively (p<0.0001), achieved at least two acceptable blows, the minimum requirement. Nonacceptability was largely ascribable to failure to satisfy end-of-test criteria; a blow of at least 6 s. Visual inspection of the results of these blows as registered on the spirometer for the presence of a plateau on the volume-time curve suggests that < 15% were acceptable. CONCLUSIONS: Although a significant training effect was demonstrated, the quality of the spirometry performed in clinical practice did not generally satisfy full ATS criteria for acceptability and reproducibility. Further study would be required to determine the clinical impact. However, the ATS guidelines allow for the use of data from unacceptable or nonreproducible maneuvers at the discretion of the interpreter. Since most of the failures were end-of-test related, the FEV1 levels are likely to be valid. Our results serve to emphasize the importance of effective training and quality assurance programs to the provision of successful spirometry in primary care practice. PMID- 10453872 TI - Oral prednisolone followed by inhaled budesonide in newly diagnosed pulmonary sarcoidosis: a double-blind, placebo-controlled multicenter study. Finnish Pulmonary Sarcoidosis Study Group. AB - STUDY OBJECTIVE: To evaluate the efficacy of oral prednisolone, followed by inhaled budesonide, in patients with newly diagnosed (<3 months) stage I and stage II pulmonary sarcoidosis. DESIGN: Double-blind, placebo-controlled, parallel-group, multicenter study. SETTING: Twenty pulmonary medicine departments in Finland. PATIENTS: One hundred eighty-nine adult patients were randomized to treatment. Patients with erythema nodosum or stage IV sarcoidosis (pulmonary fibrosis), and patients requiring immediate treatment with oral corticosteroids for extrapulmonary lesions or chronic illnesses were excluded. TREATMENT: The patients received either oral prednisolone for 3 months (20 mg/d for 8 weeks, 15 mg/d for 2 weeks, and 10 mg/d for 2 weeks) followed by inhaled budesonide (Pulmicort Turbuhaler; Astra Draco; Lund, Sweden) for 15 months at 800 microg bid, or placebo tablets followed by placebo inhaler therapy. MEASUREMENTS: Chest radiographs, lung volumes (FVC), diffusing capacity of the lung for carbon monoxide (D(LCO)), serum angiotensin-converting enzyme (SACE), and beta2 microglobulin at 3-month intervals. RESULTS: After 3 months of treatment, radiographic improvements were seen in the active-treatment group when compared to the placebo-treatment group. At 6 months, the difference was still statistically significant. Later, no differences were found. In patients with initial stage I lesions, neither the FVC nor the D(LCO) (the percent predicted mean values) changed during the study, as they were normal from the beginning. In patients with initial stage II disease, the difference in the FVC mean values between the groups also remained unchanged throughout the study. In stage II patients treated for 18 months, but not earlier, the difference in D(LCO) became statistically significant; the largest differences were seen in patients with initial FVC values <80% of predicted and D(LCO) values <75% of predicted. The decrease in SACE in the active-treated stage II patients was significantly larger than in the placebo-treated patients. No difference was observed in adverse events between the active-treated patients and the placebo-treated patients. CONCLUSION: Treatment is not required for patients with stage I disease. An initial treatment with prednisolone followed by long-term inhalation of budesonide is more effective than placebo in patients with stage II disease. Sequential oral and inhaled corticosteroid therapy may be an alternative treatment regimen for stage II sarcoidosis patients, rather than long-term oral corticosteroid therapy alone. PMID- 10453874 TI - High-frequency percussive ventilation improves oxygenation in patients with ARDS. AB - STUDY OBJECTIVES: To evaluate changes in respiratory and hemodynamic function of patients with ARDS and requiring high-frequency percussive ventilation (HFPV) after failure of conventional ventilation (CV). DESIGN: Retrospective case series. SETTING: Surgical ICU (SICU) and medical ICU (MICU) of an academic county facility. MEASUREMENTS AND RESULTS: Thirty-two consecutive patients with ARDS (20 from SICU, 12 from MICU) who were unresponsive to at least 48 h of CV and were switched to HFPV were studied. Data on respiratory and hemodynamic parameters were collected during the 48 h preceding and the 48 h after institution of HFPV and compared. Between the period of CV and the period of HFPV, the ratio of PaO2 to the fraction of inspired oxygen (F(IO2)) increased ([mean+/-SE] 130+/-8 vs. 172+/-17; p = 0.027), peak inspiratory pressure (PIP) decreased (39.5+/-1.7 vs. 32.5+/-1.9 mm Hg; p = 0.002), and mean airway pressure(MAP) increased (19.2+/-1.2 vs. 27.5+/-1.4 mm Hg; p<0.001). The rate of change of PaO2/F(IO2) per hour was also significantly improved between the two periods. The same changes in PaO2/F(IO2), PIP, and MAP were observed when the last value recorded while the patients were on CV was compared with the first value recorded after 1 h of HFPV. This improvement was sustained but not amplified during the hours of HFPV. The patterns of improvement in these three parameters were similar in SICU and MICU patients as well as in volume-control and pressure-control patients. There were no changes in hemodynamic parameters. CONCLUSION: The HFPV improves oxygenation by increasing MAP and decreasing PIP. This improvement is achieved soon after institution of HFPV and is maintained without affecting hemodynamics. PMID- 10453873 TI - Pulmonary arteriovenous malformations: screening procedures and pulmonary angiography in patients with hereditary hemorrhagic telangiectasia. AB - BACKGROUND: Hereditary hemorrhagic telangiectasia (HHT) is a dominantly inherited disease with a high prevalence of pulmonary arteriovenous malformations (PAVMs). The first symptom of HHT may be stroke or fatal hemoptysis associated with the presence of PAVM. OBJECTIVE: To evaluate different screening methods applied for the identification of PAVMs. SETTING: Odense University Hospital. SUBJECTS: HHT patients with positive findings on contrast echocardiography (CE) who participated in a screening investigation and underwent pulmonary angiography (PA). METHODS: Different screening methods were evaluated against the results of PA. In a group of patients with positive findings on CE, we compared results of PA with the following: severity of dyspnea; results of pulse oximetry arterial oxygen saturation (SaO2) supine and upright; supine PaO2 in room air and while breathing 100% oxygen; size of arteriovenous shunt in supine position; chest radiograph; and intensity of contrast at CE. RESULTS: PA was performed in 25 HHT patients with positive findings on CE, 15 of whom had PAVM. Embolization therapy was recommended in 12 patients, and 3 patients had small PAVMs not accessible for therapy. In 10 patients, PAVM could not be demonstrated at PA. The sensitivity and specificity calculated for the screening procedures are as follows: 53% and 90%, respectively, for SaO2; 60% and 100%, respectively, for chest radiograph; 73% and 80%, respectively, for PaO2 in room air; 100% and 40%, respectively, for PaO2 breathing 100% oxygen; and 64% and 80%, respectively, for shunt measurement. CONCLUSION: Initial screening with CE followed by measurement of PaO2 while breathing 100% oxygen seemed to be the best screening procedure for identification of patients with PAVM. Screening with chest radiograph and pulse oximetry was shown to be insufficient. PMID- 10453875 TI - Four-year experience with a unit for long-term ventilation (respiratory special care unit) at the Cleveland Clinic Foundation. AB - BACKGROUND: In the context that special weaning units have been advocated as effective alternatives to the ICU for weaning selected patients, we initiated a Respiratory Special Care Unit (ReSCU) at the Cleveland Clinic Hospital in August 1993. The goals of the ReSCU were the following: (1) to wean ventilator-dependent patients when possible; and (2) when weaning was not possible, to optimize patient and family instruction for patients going home with ventilatory support. This study presents our 4-year experience with 212 patients managed in the ReSCU and analyzes clinical features associated with favorable clinical outcomes. METHODS: The features of the ReSCU include six private beds in a pulmonary inpatient ward staffed by nurses with special pulmonary expertise; 24-h respiratory therapy supervision; bedside and central noninvasive monitoring (i.e., continuous pulse oximetry, end tidal capnometry, and ventilator alarms); and a multidisciplinary approach involving dietitians, physical therapists, occupational therapists, social workers, and speech pathologists. All ReSCU patients were cared for primarily by a pulmonary/critical care attending physician and fellow, with consultative input solicited as deemed necessary. The criteria for admission to the ReSCU included hemodynamic stability; absence of an arrhythmia requiring telemetry; and in the attending physician's judgment, the ability to benefit from the ReSCU. RESULTS: Between August 23, 1993, and August 31, 1997, 212 patients were admitted to the ReSCU. The median age was 68 years old; 55% were women; 86% were white; and 55% were transferred from the medical ICU. Underlying reasons for ventilator dependence were ARDS from a nonsurgical cause (33%), ARDS following surgery (18%), status post-cardiothoracic surgery (13%), status post-thoracic surgery (12%), and COPD (12%). The median length of ReSCU stay was 17 days (interquartile range, 10 to 29 days). Eighteen percent (n = 38) died during the hospitalization. Among the 174 survivors, complete ventilator independence was achieved in 127 patients (60% of the 212 patient cohort), 28 patients were ventilator dependent (13% of 212 patients), and the remaining 19 patients (9%) required partial ventilatory support. Univariate analysis regarding the association of baseline characteristics with death identified lower albumin and transferrin levels, increasing age, and the physician's estimate of lower weaning likelihood as significant correlates of death. In contrast, achieving complete ventilator independence was associated with a higher serum albumin level, a nonmedical ICU referral source, a cause of respiratory failure other than COPD, and a physician's estimate of higher weaning likelihood. To analyze the financial impact of the ReSCU, we assumed that ReSCU patients would have otherwise stayed in the medical ICU and compared the charges (ICU vs ReSCU) with, for a subset of patients, the true costs of ReSCU vs. ICU care. Analyses of both charges and cost differences showed similar savings associated with ReSCU care ($13,339 per patient [charges] and $10,694 per patient [costs]). CONCLUSIONS: We conclude the following: (1) the rate of achieving complete ventilator independence in the ReSCU was high; and (2) based on our achieving clinical outcomes, which are comparable to the most favorable rates reported in other series from ventilator units, we conclude that the ReSCU can be an effective and cost-saving alternative to the ICU for carefully selected patients. PMID- 10453876 TI - Predicting 3-day and 7-day outcomes of weaning from mechanical ventilation. AB - OBJECTIVE: To determine the correlation of acute physiology and chronic health evaluation (APACHE) II score and various weaning indexes (WIs) with 3- and 7-day weaning outcomes. DESIGN: Prospective, observational. SETTING: The medical ICU of a teaching, urban hospital. METHODS: The study included 118 adults referred for weaning from mechanical ventilation (MV). Critical care physicians, critical care nurses, and respiratory care practitioners were asked to predict whether it would take < or =3 days, 4 to 7 days, or > or =8 days to wean each patient from MV. The WIs and APACHE II scores were measured or calculated. The causes of respiratory failure, the duration of MV before initiating weaning assessment, and the 3- and 7-day weaning outcomes were obtained. Significance was set at p<0.05. RESULTS: The most common causes of respiratory failure were pneumonia (38 cases) and acute exacerbation of COPD (29 cases). Fifty-seven patients (48%) were successfully weaned from MV within 3 days of weaning assessment, and 67 (57%) were weaned within 7 days. The percentages of correct prediction of 3-day weaning outcome by critical care physicians, critical care nurses, and respiratory care practitioners were 64%, 62%, and 59%, respectively; for 7-day weaning outcome, 60%, 64%, and 58%, respectively. The successfully weaned groups had significantly lower APACHE II scores and higher maximal inspiratory pressures than the unsuccessfully weaned (failure) groups. There were no significant differences between the two groups for the remaining indexes, including rapid shallow breathing, dynamic compliance, static compliance, spontaneous respiratory rate, and the ratio of PaO2 to the fraction of inspired oxygen. CONCLUSIONS: The overall severity of illness as assessed by APACHE II score correlates better with 3- and 7-day weaning outcome than the published WIs. PMID- 10453877 TI - Role of different routes of tracheal colonization in the development of pneumonia in patients receiving mechanical ventilation. AB - STUDY OBJECTIVE: To evaluate the importance of the different pathogenic pathways involved in the development of ventilator-associated pneumonia (VAP). DESIGN: Prospective study. SETTING: An 18-bed medical and surgical ICU. PATIENTS: One hundred twenty-three patients receiving mechanical ventilation (MV). INTERVENTIONS: Tracheal, pharyngeal, and gastric samples were obtained simultaneously every 24 h. In cases where VAP was suspected clinically, bronchoscopy with protected specimen brush and BAL were performed. Semiquantitative cultures of pharyngeal samples and quantitative cultures for the remaining samples were obtained. RESULTS: Tracheal colonization at some time during MV was observed in 110 patients (89%). Eighty patients had initial colonization, 34 patients had primary colonization, and 50 patients had secondary colonization. Nineteen patients had VAP, and 25 organisms were isolated. For none of these organisms was the stomach the initial site of colonization. Gram positive organisms colonized mainly in the trachea during the first 24 h of MV (p<0.001). On the contrary, enteric Gram-negative bacilli (p<0.001) and yeasts (p<0.002) colonized the trachea secondarily. Previous endotracheal intubation (p<0.005) and acute renal failure before admission to the ICU (p<0.001) were associated with colonization by Pseudomonas aeruginosa; prior antibiotics were associated with colonization by Acinetobacter baumanii (p<0.05) and yeasts (p<0.006); and cranial trauma was associated with Staphylococcus aureus colonization (p<0.035). CONCLUSIONS: Although the stomach can be a source of organisms that colonize the tracheobronchial tree, it is a much less common source of the bacteria that cause VAP. The pattern of colonization and risk factors may be different according to the type of organisms involved. PMID- 10453878 TI - Hyperlactatemia and pulmonary lactate production in patients with fulminant hepatic failure. AB - STUDY OBJECTIVES: To determine whether the lungs of patients with fulminant hepatic failure release lactate, and if so, whether this release relates to systemic lactate concentration or acid base status. Another objective was to examine the accuracy of lactate flux calculations in critically ill patients. DESIGN: Prospective observational study. SETTING: The ICU of a major teaching hospital. PATIENTS: Twelve patients with fulminant hepatic failure; 30 other critically ill patients in whom a pulmonary artery catheter was in place. INTERVENTIONS: None. MEASUREMENT AND RESULTS: The precision of whole-blood lactate measurements was assessed in 30 patients with critical illnesses of variable etiology who had a wide range of arterial lactate concentrations. The reliability of lactate measurements decreased with increasing lactate concentration. In each patient with liver failure, pulmonary lactate flux was calculated on three occasions using the Fick principle. Arterial blood lactate concentration was consistently higher than venous concentrations, indicating lactate release by the lungs (mean difference, 0.15 mmol/L; 95% confidence interval, 0.09 to 0.21; p<0.001). Mean pulmonary lactate production for the 12 patients was 83 mmol/h (range, 22 to 210 mmol/h). No patient had significant acute lung injury. Correlations were found among the arterial lactate concentration and both the arteriovenous (AV) lactate difference (p<0.025) and pulmonary lactate production (p<0.05), but not with acid-base status or cardiac output. The reliability of individual AV lactate difference calculations and pulmonary lactate flux calculations was poor. CONCLUSION: The lungs release lactate in patients with fulminant hepatic failure at a rate proportional to the degree of systemic hyperlactatemia. However, the measurement errors associated with pulmonary lactate flux calculations using the Fick principle are large, so individual measurements should be interpreted with caution. PMID- 10453879 TI - Proliferation of type II pneumocytes and alteration in their apical surface membrane antigenicity in pulmonary sarcoidosis. AB - STUDY OBJECTIVE: To evaluate both the proliferation of type II pneumocytes in the alveolitis associated with pulmonary sarcoidosis and any alteration in their surface membrane antigenicity. MATERIALS AND METHODS: We investigated 20 transbronchial lung biopsy (TBLB) specimens from 20 patients with pulmonary sarcoidosis, 7 TBLB specimens from 7 sarcoidosis patients without pulmonary involvement, and 19 normal lung specimens, using colloidal iron stain and immunostaining with anti-Thomsen-Friedenreich (TF) antigen and anti-surfactant protein-A monoclonal antibodies. RESULTS: The density of type II pneumocytes was significantly higher in the pulmonary sarcoidosis specimens ([mean +/- SD] 11.1+/ 3.7 per 1 mm alveolar septal length) than in the nonpulmonary sarcoidosis (7.8+/ 1.3) or normal lung specimens (7.2+/-0.8). TF antigen was directly expressed on the apical surface of some type II pneumocytes in the pulmonary sarcoidosis specimens, but it was completely masked by sialic acids in the nonpulmonary sarcoidosis specimens and in the normal lung tissues. CONCLUSIONS: In pulmonary sarcoidosis, type II pneumocytes proliferated and the antigenicity of the surface membrane was altered. It is suggested that these type II pneumocytes may be vulnerable to injury by natural anti-TF antibodies that are cytotoxic when present with complement. This damage may decrease alveolar surfactant and cause focal alveolar collapse proceeding to pulmonary fibrosis in some cases of pulmonary sarcoidosis. PMID- 10453880 TI - Captopril lowers coronary venous free radical concentration after direct current cardiac shocks. AB - STUDY OBJECTIVES: Direct current (DC) shocks to the heart cause morphologic and functional myocardial damage. Previous studies have suggested that acute DC shock injury is free radical mediated and that the administration of antioxidant enzymes superoxide dismutase and catalase can reduce the level of DC shock induced free radicals. Angiotensin-converting enzyme (ACE) inhibitors are clinically used drugs that may scavenge free radicals or reduce free radical generation. The objective of our study was to determine whether the ACE inhibitor captopril lowers free radicals after DC shocks. DESIGN: In six open-chest dogs, we administered 100-J DC shocks to the epicardium, before and after administration of captopril, 3 mg/kg. We used electron paramagnetic resonance measurements of arterial and coronary venous ascorbate free radical (AFR) as a real-time marker of free radical generation (total oxidative flux). MEASUREMENTS AND RESULTS: Captopril resulted in a significant lowering of coronary venous AFR concentration: the peak rise in AFR after 100-J shocks was 17.3+/-3.4% (mean +/- SEM before captopril vs. 3.2+/-4.0% after captopril; p<0.05). CONCLUSIONS: Captopril lowers coronary venous AFR concentration after high-energy epicardial shocks. PMID- 10453881 TI - Emerging concepts in the evaluation of ventilatory limitation during exercise: the exercise tidal flow-volume loop. AB - Traditionally, ventilatory limitation (constraint) during exercise has been determined by measuring the ventilatory reserve or how close the minute ventilation (VE) achieved during exercise (i.e., ventilatory demand) approaches the maximal voluntary ventilation (MVV) or some estimate of the MVV (i.e., ventilatory capacity). More recently, it has become clear that rarely is the MVV breathing pattern adopted during exercise and that the VE/MVV relationship tells little about the specific reason(s) for ventilatory constraint. Although it is not a new concept, by measuring the tidal exercise flow-volume (FV) loops (extFVLs) obtained during exercise and plotting them according to a measured end expiratory lung volume (EELV) within the maximal FV envelope (MFVL), more specific information is provided on the sources (and degree) of ventilatory constraint. This includes the extent of expiratory flow limitation, inspiratory flow reserve, alterations in the regulation of EELV (dynamic hyperinflation), end inspiratory lung volume relative to total lung capacity (or tidal volume/inspiratory capacity), and a proposed estimate of ventilatory capacity based on the shape of the MFVL and the breathing pattern adopted during exercise. By assessing these types of changes, the degree of ventilatory constraint can be quantified and a more thorough interpretation of the cardiopulmonary exercise response is possible. This review will focus on the potential role of plotting the extFVL within the MFVL for determination of ventilatory constraint during exercise in the clinical setting. Important physiologic concepts, measurements, and limitations obtained from this type of analysis will be defined and discussed. PMID- 10453883 TI - Clinical indications for noninvasive positive pressure ventilation in chronic respiratory failure due to restrictive lung disease, COPD, and nocturnal hypoventilation--a consensus conference report. PMID- 10453882 TI - Advances in the treatment of malignant pleural mesothelioma. AB - Malignant pleural mesothelioma is a neoplasm that is commonly fatal and for which there are no widely accepted curative approaches. Mesothelioma is unresponsive to most chemotherapy and radiotherapy regimens, and it typically recurs even after the most aggressive attempts at surgical resection. Multimodality approaches have been of some benefit in prolonging survival of very highly selected subgroups of patients, but they have had a relatively small impact on the majority of the patients diagnosed with this disease. As the incidence of pleural mesothelioma peaks in the United States and Europe over the next 10 to 20 years, new therapeutic measures will be necessary. This review will discuss the roles of chemotherapy, radiotherapy, surgery, and combined modality approaches in the treatment of pleural mesothelioma, as well as scientific advances made in the past decade that have led to the development of experimental techniques, such as photodynamic therapy, immunotherapy, and gene therapy, that are currently undergoing human clinical trials. These promising new avenues may modify the therapeutic nihilism that is rampant among clinicians dealing with mesothelioma. PMID- 10453884 TI - The significance of serum vs tissue levels of antibiotics in the treatment of penicillin-resistant Streptococcus pneumoniae and community-acquired pneumonia: are we looking in the wrong place? PMID- 10453885 TI - Is meta-analysis really meta-physics? PMID- 10453886 TI - Aerosol-derived airway morphometry and aerosol bolus dispersion in patients with lung fibrosis and lung emphysema. AB - OBJECTIVE: Patients with lung emphysema show increased aerosol-derived dimensions of peripheral airspaces and increased aerosol bolus dispersion (AD). To apply these tests in epidemiologic studies, the objective of this pilot study was to investigate whether morphometric changes caused by lung fibrosis can be distinguished from those caused by emphysema. DESIGN: This study was designed as a cross-sectional study in which airspace dimensions and AD in patients with emphysema and in patients with fibrosis were compared. Forty patients participated in the study: 20 patients had high-resolution CT (HRCT)-proved lung emphysema and 20 patients had HRCT-proved lung fibrosis. All patients underwent conventional lung function tests, aerosol-derived airway morphometry (ADAM), and AD measurements. RESULTS: Patients with lung emphysema showed normal dimensions of small airways but enlarged airspace dimensions in the lung periphery. Patients with fibrosis showed in all lung depths increased airspace dimensions. AD was increased in patients with emphysema but was normal in patients with fibrosis. CONCLUSIONS: These results show that when using ADAM and AD, morphometric changes caused by emphysema can be distinguished from those caused by fibrosis with high sensitivity and specificity. PMID- 10453887 TI - Clinical conference on management dilemmas: bullous hand pustules in a patient treated for lung cancer. PMID- 10453888 TI - A 27-year-old woman with cough, dyspnea, and wheezing. PMID- 10453889 TI - Pulmonary hypertension in a 45-year-old woman with recurrent deep venous thromboses. PMID- 10453890 TI - Anastomotic pulmonary hypertension after lung transplantation for primary pulmonary hypertension: report of surgical correction. AB - This report describes a patient who developed pulmonary hypertension 6 years after lung transplantation for primary pulmonary hypertension (PPH). Evaluation with right heart catheterization followed by pulmonary angiography, however, demonstrated that the pulmonary hypertension was secondary to an anastomotic narrowing of the pulmonary artery, rather than a recurrence of her PPH. Vascular complications of lung transplantation should be considered in patients who experience exertional dyspnea after lung transplantation. The suggestion of pulmonary hypertension on echocardiography should prompt further evaluation, including meticulous hemodynamic measurements. PMID- 10453891 TI - Development of nonspecific interstitial pneumonitis associated with long-term treatment of primary pulmonary hypertension with prostacyclin. AB - A young woman with primary pulmonary hypertension presented with interstitial lung disease approximately 5 years after successful treatment with IV prostacyclin. The pathology was consistent with nonspecific interstitial pneumonitis and was unresponsive to steroids and immunosuppressive medications. We speculate that further cases of this syndrome may be reported as more patients are living beyond 5 years with prostacyclin. PMID- 10453892 TI - Interferon-alpha therapy associated with the development of sarcoidosis. AB - Interferons (IFNs) have been implicated in the pathogenesis of sarcoidosis. In particular, IFN-gamma has been linked to pulmonary macrophage activation, a characteristic feature of sarcoidosis. IFN-alpha is now being administered therapeutically in a variety of conditions. To date, IFN-alpha has not been implicated in the pathogenesis of sarcoidosis. We report the case of a 50-year old woman who developed sarcoidosis while being treated with IFN-alpha for chronic myelogenous leukemia. Her disease activity correlated with the dosage of IFN-alpha. We speculate that the immunomodulatory effects of IFN-alpha triggered clinical manifestations of sarcoidosis in this patient. PMID- 10453893 TI - Disseminated Pseudallescheria boydii infection in a nonimmunocompromised host. AB - We present a highly unusual case of pulmonary Pseudallescheria boydii infection in a nonimmunocompromised host with a cavitating mass lesion. The diagnosis was confirmed by open lung biopsy. The patient was treated at another institution with course of amphotericin B, considered an ineffective therapy for this infection, and presented to us with direct extension and invasion of the left atrial appendage and the pulmonary artery, followed by massive pulmonary embolization and hematogenous dissemination to the liver, spleen, kidney, pancreas, and brain. PMID- 10453894 TI - Myocardial bridging as a cause of acute transient left heart dysfunction. AB - The significance of myocardial bridging is still a matter of debate, and although several reports have underlined its pathologic potential, myocardial bridging is often considered to be a benign phenomenon. We present here the case of a 63-year old woman with a history of acute left heart failure and ECG evidence of ischemia, and whose primary abnormality on extensive workup was myocardial bridging. This case further underlines that myocardial bridging can lead to significant cardiac events. PMID- 10453895 TI - Insulin by inhalation? PMID- 10453896 TI - Pulmonary administration of insulin as an aerosol. PMID- 10453897 TI - Herbalism for the treatment of asthma. PMID- 10453898 TI - Sialyl Lewis X-i antigen in pleural effusion. PMID- 10453899 TI - Fungal beta-glucan can yield false-positive results with the limulus amebocyte lysate endotoxin assay. PMID- 10453900 TI - Pleurodural fistulas and neurologic manifestations. PMID- 10453901 TI - Research in developing countries. PMID- 10453902 TI - Low- vs high-dose inhaled albuterol for the treatment of acute asthma. PMID- 10453903 TI - Intentional carbon monoxide poisoning. PMID- 10453904 TI - "Diagnostic" pulmonary function tests. PMID- 10453905 TI - Streptokinase for endobronchial blood clots. PMID- 10453906 TI - Reversible unilateral diaphragmatic paralysis in pulmonary embolism. PMID- 10453907 TI - Bendectin: review of the medical literature of a comprehensively studied human nonteratogen and the most prevalent tortogen-litigen. PMID- 10453908 TI - Dr. Brent and scientific debate. PMID- 10453909 TI - Response to Dr. Stuart Newman's commentary on an article entitled "Bendectin: review of the medical literature of a comprehensively studied human nonteratogen and the most prevalent tortogen-litigen". PMID- 10453911 TI - Cadmium affects testes and semen of rabbits exposed before and after puberty. AB - Semen quality and testicular characteristics were measured in 92 rabbits in three controlled experiments with males exposed to cadmium chloride (Cd) when 12 or 27 weeks old. Doses of Cd were administered subcutaneously (s.c.), orally, or intravenously (i.v.) and subsets of animals were unilaterally castrated or not to evaluate the testes and to collect semen repeatedly from males when adults. There was considerable variability but Cd given at 12 weeks of age in doses of 0.08 mmol/kg s.c., 0.20 mmol/kg orally, and 0.02 mmol/kg i.v. tended to depress sperm output of these males when adults. The 0.02 mmol/kg dose given i.v., 0.4 mmol/kg orally, and 0.16 mmol/kg s.c. were lethal to many animals. Treatment of adults resulted in a generally similar pattern of systemic toxicity, and limited comparisons suggested that testicular sensitivity was slightly less than for young males. Androgenic function usually was maintained, as indicated by normal libido and seminal volumes even in males with reduced spermatogenesis. Necropsies confirmed previous findings of hyperemia, hemorrhaging, necrosis, and destruction of all spermatogenic elements in severely affected males. PMID- 10453910 TI - Influence of environmental cadmium on testicular proliferation in roe deer. AB - The influence of cadmium on spermatogenesis in roe deer was studied, comparing animals from areas with high (n = 37) and low (n = 23) cadmium exposure. Testes were evaluated during pre-rut (May) and rut (July/August). During these periods the cadmium accumulation (mg/kg kidney) in polluted regions averaged 3.41 and 3.52 in comparison with 1.61 and 1.81 in controls. Cadmium accumulation was positively correlated with testis weight during both seasons. In May the proliferation (units of tissue polypeptid specific antigen/g parenchyma) was lower in animals with higher cadmium (24.02 compared with 78.20; P < 0.01). During the rut, testicular proliferation and spermatozoa/g testis as well as apoptosis showed no significant changes with increased cadmium contamination. The results suggest delayed proliferation during the pre-rutting period in animals with high cadmium exposure, but other indications of effects on the testis were not significant. PMID- 10453912 TI - Epididymal sperm motion as a parameter of male reproductive toxicity: sperm motion, fertility, and histopathology in ethinylestradiol-treated rats. AB - The present study was designed to characterize the effect of ethinylestradiol (EE) on epididymal sperm motion using a computer-assisted sperm analysis system (CASA), and to elucidate the correlation between sperm motion endpoints and other measures including fertility, histopathologic, and endocrinologic endpoints. EE was orally given to adult male rats at a daily dosage of 10 mg/kg for 3 and 5 d, and at daily dosages of I and 10 mg/kg for 1, 2, 3, and 4 weeks. Changes in sperm motion were first detected after one week of treatment. Of nine sperm motion parameters, the percentage of motile sperm, velocity, and amplitude of the lateral head displacement (ALH) were decreased in the 10 mg/kg dosing group. Accompanying the decreases in those parameters, the male fertility indices in the 10 mg/kg dosing group were reduced after one week of treatment, and no males in this group could impregnate intact females after 2 weeks or more of treatment. The number of sperm heads in the cauda epididymis in the 10 mg/kg dosing group was reduced to about one-half that in the control group after one week of treatment, whereas the total number of homogenization-resistant advanced spermatids in the testis was not altered and only a slight change was detected in the number and morphology of germ cells in the testis. These results suggest that reduction in the number of epididymal sperm and in sperm motion are not secondary to testicular alteration. However, after 3 weeks of treatment, the number of sperm heads in the testis was drastically reduced with severe atrophy of the seminiferous tubules both in the 1 and 10 mg/kg dosing groups. The profiling of epididymal luminal fluid proteins indicated that two major bands that migrated with molecular weights of about 22 and 23 kDa were weakened and their density was reduced to approximately 70% of the control after 5-d and one week treatments in the 10 mg/kg dosing group. Circulating testosterone declined drastically after 3 d of treatment and remained at undetectable levels with a concomitant decline of circulating LH and FSH, suggesting that EE inhibits testosterone secretion immediately via a negative feedback system, and there follow changes in the accessory reproductive organs including the epididymis. These results indicate that EE affects epididymal spermatozoa before testicular germ cells via a testosterone deficiency, when it is administered at extremely high dosages. The reduction in the sperm motion manifested as decreases in the percentage of motile sperm, ALH, and velocity, is considered to be responsible for the onset of infertility. Sperm motion analysis could be particularly useful for detecting the toxic effects of chemicals that act through the endocrinologic system on the epididymis. PMID- 10453913 TI - Cytotoxic effect of endosulfan on rat Sertoli-germ cell coculture. AB - Endosulfan induced testicular impairment has been reported in vivo in rats. The present study was conducted to evaluate the cytotoxic effects of endosulfan in vitro employing rat testicular cells in culture, that is Sertoli-germ cell coculture. Cytotoxic changes induced by endosulfan (0, 2, 20, 40, and 80 microM) in mixed cultures of Sertoli and germ cells were seen after 24 and 48 h of treatment. Endosulfan led to an increase in germ cell detachment from the Sertoli cell monolayer in a dose dependent manner. A loss in the viability of detached cells was observed in all treated groups. Substantial leakage of the cytosolic enzyme lactate dehydrogenase was observed in the medium after 24 and 48 h endosulfan treatment at concentrations of 20 to 80 microM. The extent of toxicity was greater after 48 h of treatment. PMID- 10453914 TI - Developmental toxicity in rat fetuses exposed to the benzimidazole netobimin. AB - Netobimin (NTB) is a prodrug of albendazole (ABZ) and is used as a broad-spectrum anthelmintic both in human and veterinary medicine. Pregnant Sprague-Dawley rats were treated po with 50, 59.5 and 70.7 mg/kg of NTB on Gestational Day (GD) 10. The results, observed on GD 20, demonstrated that NTB induced a significant increase of resorptions. Moreover, decreased fetal body weight and an increase in skeletal malformations were observed in treated groups. We report the first study in which vascular malformations are described in rats after the administration of a benzimidazole compound. An interesting relationship between intercostal vessel and rib malformations was found. PMID- 10453915 TI - Reproductive function in rats exposed neonatally to bisphenol A and estradiol benzoate. AB - The reproductive function in rats treated subcutaneously (s.c.) with 300 microg/g bisphenol A or 2 microg/g estradiol benzoate from postnatal Day 1 to 5 was examined after puberty as well as histolopathogic changes in reproductive organs. All male and female rats treated postnatally with estradiol benzoate showed poor reproductive capability, including adverse effects on masculine sexual behavior, and marked histopathologic alterations of the reproductive organs. In addition, estradiol benzoate markedly reduced the volume of the sexually dimorphic nucleus of the preoptic area (SDN-POA) in males. On the other hand, all male and female rats treated postnatally with bisphenol A showed normal reproductive function and no histopathologic abnormalities of reproductive organs. Bisphenol A did not affect the volume of the SDN-POA. These results indicated that neonatal exposure to estradiol benzoate affects reproductive function in male and female rats, and treatment with bisphenol A at a fairly high dose was ineffective if given postnatally to male and female rats. PMID- 10453916 TI - The perception of teratogenic risk by women with nausea and vomiting of pregnancy. AB - This study determined the advice reported to be received by women suffering from nausea and vomiting of pregnancy (NVP) from their caregivers regarding management, the teratogenic risk perception of these women and their choice of antiemetic drug use in pregnancy. A secondary objective was to determine prospectively the effect of counseling on malformation risk perception in women with NVP. The women were prospectively followed-up and questioned about the use of pharmacotherapy or other management choices as well as their perception of teratogenic risk through structured telephone interviews. The results showed that at the initial call, around 6 weeks of gestation, over three quarters of the 260 participants reported that therapy of NVP increased their teratogenic risk. This risk perception was decreased significantly after counseling. Women who reported their physicians' advice to change their diet and/or lifestyle attributed an increased risk for major malformations with antiemetics for NVP (P = 0.001), whereas women who reported advice to take antiemetic medications known to be safe to the fetus attributed no change in risk for major malformations with drugs for NVP (P = 0.002). We came to the conclusion that women are commonly hesitant to treat NVP pharmacologically due to unfounded fears of teratogenic risk. Evidenced based counseling resulted in reduced numbers of women who considered drug therapy for NVP to increase the risk of major malformations. PMID- 10453917 TI - Relationship between mechanomyogram and force during voluntary contractions reinvestigated using spectral decomposition. AB - A mechanomyogram (MMG) is considered to represent the pressure waves resulting from the lateral expansion of contracting muscle fibers. However, the actual MMG recording appears not only to reflect lateral changes of active fibers, but also to include the effect of their longitudinal shortening, because the fiber orientation, particularly in pennate muscles, is not parallel with the MMG transducer attached at the skin surface. In the present investigation, a spectral decomposition method was developed to eliminate the interference due to fiber longitudinal movement from the MMG recording. The MMG was recorded over the belly of the rectus femoris muscle, which is a pennate muscle. Vibration over the tibial tuberosity (VTT) was used as a measure of the integrated longitudinal movement of the muscle fibers. The lateral and longitudinal components included in the MMG were separated by a spectral decomposition method that is based on the coherence function of the MMG and VTT. The MMG/force relationship was compared between the original and decomposed MMG. One-third of the 12 subjects demonstrated a curvilinear relationship between the original MMG and force throughout the range of force. In the other two-thirds, the MMG saturated or reduced beyond 70% of the maximum voluntary contraction (MVC). After decomposition, the MMG increased progressively with force up to 70% MVC, beyond which it decreased in all subjects. The spectral decomposition method described here is considered to be a useful tool with which to examine in more detail the MMG/force relationship of different pennate muscles. PMID- 10453918 TI - Estrogen attenuates HSP 72 expression in acutely exercised male rodents. AB - Estrogen has been shown to reduce post-exercise skeletal muscle damage. Exercise induced muscle damage may be a factor in the elevated post-exercise expression of heat-shock proteins (HSPs). Thus, the present investigation was conducted in order to examine the influence of estrogen on post-exercise levels of HSP 72 and heat-shock cognate, HSC 73, in male and female rodents. Prior to an acute bout of treadmill running, male and female Sprague-Dawley rats received daily injections of either 40 microg x kg(-1) of beta-estradiol 3-benzoate or olive oil vehicle for 2 weeks. A two- to fourfold reduction in post-exercise HSP 72 content was observed in the heart, liver, lung and red and white vastus muscles of estradiol treated males compared with their vehicle-injected counterparts (P < 0.05). Compared to the males, the females had significantly lower post-exercise HSP 72 levels which were not affected by estradiol supplementation. Moreover, estradiol administration in male rodents resulted in a HSP response similar to that of females following exercise. Thus, the results of the present investigation suggest that estrogen is the factor responsible for the observed differences in post-exercise HSP 72 levels between males and females. PMID- 10453919 TI - A comparison of electrical activity in the triceps surae at maximum isometric contraction with the knee and ankle at various angles. AB - The purpose of this study was to test the endurance of the soleus muscle, and to examine the joint position at which it is most active, while simultaneously suppressing the activity of the gastrocnemius. Ten young males performed maximum isometric contraction of the triceps surae for 100 s, and the endurance and plantar flexion torque of this muscle were measured at various angles of the knee and ankle joints. The electromyogram was measured simultaneously and subsequently converted into integrated electromyogram (IEMG) values. With the knee flexed at 130 degrees, the rate of change in IEMG values for the soleus (0.454% x s(-1)) with the ankle in a neutral position was significantly higher than that for the medial and lateral gastrocnemius. Both with the ankle dorsiflexed at 10 degrees and in the neutral position, the rate of change in IEMG for the soleus was significantly higher with the knee flexed at 90 degrees and 130 degrees than with the knee fully extended. With the knee flexed at 90 degrees and 130 degrees, the IEMG activity of the soleus during the initial (5-10 s) and final 5 s tended to be higher than those for the medial and lateral gastrocnemius, regardless of the ankle joint position. We conclude that the position in which the soleus acts most selectively during a sustained maximum isometric contraction of the triceps surae is with the ankle in a neutral position and the knee flexed at 130 degrees. PMID- 10453920 TI - Relative contribution of arms and legs in humans to propulsion in 25-m sprint front-crawl swimming. AB - Eight male subjects were asked to swim 25 m at maximal velocity while the use of the arm(s) and legs was alternately restricted. Four situations were examined using one arm (1A), two arms (2A), one arm and two legs (1A2L) and both arms and legs (2A2L, normal swim) for propulsion. A significant mean increase of 10% on maximal velocity was obtained in 1A2L and 2A2L compared to 1A and 2A. A non significant 4% effect was obtained in 1A. This study focused on the actual contribution of leg kick in the 10% gain in maximal velocity. It was clear that the underwater trajectory of the wrist was modified by the action of the legs (most comparisons P < 0.001). Therefore it was thought that the legs enhanced the generated propulsive force by improving the propulsive action of the arm. The arm action was quantified by selecting typical phases from the filmed trajectory of the wrist, namely forward (F), downwards (D) and backwards (B). Although there was a tendency for individual changes in kinematic parameters (F, D and B) to occur with individual changes in velocity when 2A was compared to 2A2L, no relationship was found between the relative changes in F, D and B and relative changes in velocity. This was illustrated by describing the responses of three individuals who could represent three patterns of contribution by legs and arms to propulsion in high speed swimming. PMID- 10453921 TI - The use of Doppler and atrioventricular plane motion echocardiography for the detection of changes in left ventricular function after training. AB - In this study we compared the efficacy of Doppler and atrioventricular plane motion echocardiography in detecting the changes in left ventricular function caused by moderate-intensity training. Fifty-nine healthy men and women (aged 40 68 years) were divided into either a group of walkers (n = 32) or controls (n = 27). Pre-intervention, there were no significant differences between the groups for gender, age, height, mass or predicted maximal oxygen consumption. The walkers completed a progressive 18-week walking programme that resulted in an estimated mean gross energy expenditure whilst walking of 4.0 (1.3) MJ x week(-1) for the duration of the study, and 5.9 (1.7) MJ x week(-1) during the final 6 weeks. After the 18-week programme there were no significant changes in Doppler measures of early or late filling velocities. However, the walkers showed an increase in the velocity of relaxation (1.2 cm x s(-1)) (P < 0.02) of the left free wall, as measured using atrioventricular plane motion echocardiography, while the controls showed no significant changes. The findings suggest that atrioventricular plane motion echocardiography is more sensitive than Doppler echocardiography in detecting the left ventricular changes caused by exercise intervention. PMID- 10453922 TI - Cardiovascular responses to an orthostatic challenge and electrical-stimulation induced leg muscle contractions in individuals with paraplegia. AB - The purpose of this study was to investigate the cardiovascular and haemodynamic responses that occur during moderate orthostatic challenge in people with paraplegia, and the effect of electrical stimulation (ES)-induced leg muscle contractions on their responses to orthostatic challenge. Eight males with complete spinal lesions between the 5th and 12th thoracic vertebrae (PARA) and eight able-bodied individuals (AB) volunteered for this study. Changes in heart rate (fc), stroke volume (SV), cardiac output (Qc), mean arterial pressure (MAP), total peripheral resistance (TPR), limb volumes and indices of neural modulation of fc, [parasympathetic (PNS) and sympathetic (SNS) nervous system indicators] were assessed during: (1) supine rest (REST), (2) REST with lower-body negative pressure at -30 torr (LBNP -30, where 1 torr = 133.32 N/m2), and (3) for PARA only, LBNP -30 with ES-induced leg muscle contractions (LBNP + ES). LBNP -30 elicited a decrease in SV (by 23% and 22%), Qc (by 15% and 18%) and the PNS indicator, but an increase in fc (by 10% and 9%), TPR (by 23% and 17%) and calf volume (by 1.51% and 4.04%) in both PARA and AB subjects, respectively. The SNS indicator was increased in the AB group only. Compared to LBNP -30, LBNP + ES increased SV (by 20%) and Qc (by 16%), and decreased TPR (by 12%) in the PARA group. MAP was unchanged from REST during all trials, for both groups. The orthostatic challenge induced by LBNP -30 elicited similar cardiovascular adaptations in PARA and AB subjects. ES-induced muscle contractions during LBNP 30 augmented the cardiovascular responses exhibited by the PARA group, probably via reactivation of the skeletal muscle pump and improved venous return. PMID- 10453923 TI - Effect of exercise modality on oxygen uptake kinetics during heavy exercise. AB - The mechanisms responsible for the oxygen uptake (VO2) slow component during high intensity exercise have yet to be established. In order to explore the possibility that the VO2 slow component is related to the muscle contraction regimen used, we examined the pulmonary VO2 kinetics during constant-load treadmill and cycle exercise at an exercise intensity that produced the same level of lactacidaemia for both exercise modes. Eight healthy subjects, aged 22 37 years, completed incremental exercise tests to exhaustion on both a cycle ergometer and a treadmill for the determination of the ventilatory threshold (defined as the lactate threshold, Th1a) and maximum VO2 (VO2max). Subsequently, the subjects completed two "square-wave" transitions from rest to a running speed or power output that required a VO2 that was halfway between the mode-specific Th1a and VO2max. Arterialised blood lactate concentration was determined immediately before and after each transition. The VO2 responses to the two transitions for each exercise mode were time-aligned and averaged. The increase in blood lactate concentration produced by the transitions was not significantly different between cycling [mean (SD) 5.9 (1.5) mM] and running [5.5 (1.6) mM]. The increase in VO2 between 3 and 6 min of exercise; (i.e. the slow component) was significantly greater in cycling than in running, both in absolute terms [290 (102) vs 200 (45) ml x min(-1); P<0.05] and as a proportion of the total VO2 response above baseline [10 (3)% vs 6 (1)%; P < 0.05]. These data indicate that: (a) a VO2 slow component does exist for high-intensity treadmill running, and (b) the magnitude of the slow component is less for running than for cycling at equivalent levels of lactacidaemia. The greater slow component observed in cycling compared to running may be related to differences in the muscle contraction regimen that is required for the two exercise modes. PMID- 10453924 TI - Cardiovascular responses to light isometric and aerobic exercise in 21- and 59 year-old males. AB - The effects of aging on the cardiovascular response to continuous light isometric and aerobic exercise remains to be determined. Thus, the purpose of this study was to compare the cardiovascular response of young and older males during light handgrip and cycle ergometry exercise. Blood pressure, heart rate, rate pressure product, as well as pre-ejection period (derived from impedance cardiography) were obtained for 15 young [mean (SE) age: 21 (0.7) years] and 15 older males [59 (0.8) years] during and after light handgrip exercise and cycle ergometry. The parasympathetic influence on the heart was also assessed through a time-series analysis of heart period variability (HPVts). Both during and when recovering from the handgrip exercise and cycle ergometry, the older subjects exhibited a significantly higher absolute systolic and diastolic blood pressure, and rate pressure product, and a lower HRVts than the young subjects. Relative to baseline, the change in pre-ejection period was lower for the young subjects during the handgrip and cycle ergometry, tasks. These results indicate that although the sympathetic influence on both the myocardium and the vasculature was less pronounced in the older males, the aging cardiovasculature was under greater hemodynamic stress both during rest and during exposure to light isometric and aerobic challenge. PMID- 10453925 TI - Effects of a low-intensity conditioning programme on VO2max and maximal instantaneous peak power in elderly women. AB - The effects of 12 weeks of a low-intensity general conditioning programme on maximal instantaneous peak power (Wpeak) and maximal oxygen uptake (VO2max) were examined in 20 elderly women. After medical, familiarisation, and ethical procedures, the subjects were randomly divided into either a training and or a control group. The training group [n = 11; mean (SD) age 63.0 (3.1) years] agreed to take part in a 12-week training programme at an exercise intensity kept under 60% of the heart rate reserve for about 60 min, 3 times a week. The control group [n = 9; mean (SD) age 63.5 (3.3) years] did not perform any particular physical training. Before and after the training period, all participants underwent anthropometric measures and a maximal cycling test to exhaustion to measure their VO2max. In addition, Wpeak was determined 1 week later by the subjects performing a vertical jump from a squatting position on a force platform. Following training, neither the anthropometric characteristics nor the VO2max changed in either of the groups. In contrast, Wpeak increased significantly (P < 0.001) in the training group, but did not change in the control group. This result could be interpreted as the result of an improved level of neuromuscular activation. Furthermore, it shows that although muscle power declines with age at a faster rate than does aerobic power, its sensitivity to training seems to be higher than that of the aerobic system. PMID- 10453926 TI - Substrate utilization during submaximal exercise in obese and normal-weight women. AB - In this study we compared substrate use at submaximal intensities of a maximal graded exercise test (GXT) with that derived from equivalent intensities during continuous submaximal steady-state exercise in obese and normal-weight women. Sedentary obese (n = 20, body fat > 30%) and normal-weight (n = 15, body fat < or =30%) women performed three treadmill tests with concurrent metabolic measurements. Maximal oxygen consumption (VO2max) was determined using the Bruce protocol, followed by two, randomly assigned, continuous 15-min, steady-state exercise bouts, on different days; one bout at 50% and one bout at 75% VO2max. Analysis of variance revealed no significant differences between groups for blood lactate or respiratory exchange ratio (R) values at any point during exercise. Therefore, obese and normal-weight group data were combined for subsequent analyses. The R at 50% VO2max from the GXT [0.83 (0.01)] was significantly (P < 0.05) lower than at 8 min [0.90 (0.01)] and 15 min [0.89 (0.01)] of steady-state exercise, whereas at 75% VO2max, the GXT R [0.96 (0.01)] was similar to that seen at 8 min [0.96 (0.01)] and at 15 min of steady-state exercise [0.93 (0.01)]. Blood lactate values at 50% VO2max were similar between the GXT [1.66 (0.10) mM] and steady-state exercise [1.65 (0.09) mM], but at 75% VO2max the GXT blood lactate values [2.58 (0.21) mM] were lower than after 15 min of steady-state exercise [4.65 (0.46) mM]. Total exercise fat oxidation was greater at 50% compared to 75% VO2max. There was no difference in substrate use between sedentary obese and normal-weight women either at rest or during steady-state exercise at the same relative intensity. Total fat oxidation was greater during low- (50% VO2max) compared to high-intensity (75% VO2max) exercise. Data from a GXT cannot be used to predict R or substrate utilization values for the purpose of exercise prescription. PMID- 10453927 TI - Allometric scaling of isokinetic peak torque: the Nebraska Wrestling Study. AB - Allometric scaling has been used increasingly in the exercise sciences to control statistically for body size differences in physical performance variables. The purpose of this study was to use multivariate allometric scaling to examine the influence of fat-free mass (FFM) on age-related differences in strength in young club (8-13 years) and high-school (14-18 years) wrestlers. The dependent variables were log-transformed values of isokinetic peak torque for leg extension and flexion at 0.52, 3.14, and 5.24 rad x s(-1)(30, 180, and 300 x s(-1)). The independent variables used in the multiple regression analyses were log transformed values for FFM, age, and the FFM versus age interaction. The resulting regression equations were of the form: log Y = log a + b1 log X1 + b2 log X2 + bn log Xn. The initial multiple regression analyses showed significant interaction effects (P < 0.05) for all dependent variables, therefore separate regression analyses were performed for the younger and older groups of wrestlers. The results indicate that for the younger wrestlers there were increases in isokinetic peak torque at all velocities across age after controlling for FFM. The FFM scaling exponents ranged from 0.94 to 1.31. All exponents included 1.0 in the 95% confidence interval, except for extension at 3.14 rad x s(-1). For the high-school wrestlers, both FFM and age were significant for the extension data, but only FFM was significant for the flexion data. All FFM exponents included 1.0 in the 95% confidence interval. These results indicate that the relationship between FFM and peak torque differed across age. In addition, with the exception of the flexion data for the high-school wrestlers, within each group increases in isokinetic peak torque occurred across age, independent of increases in FFM. The causes of the age effect for strength are speculative, but it may be due to developmental changes in neuromuscular function, alterations in the distribution of muscle mass as a percentage of FFM and/or the distribution of FFM across body segments. PMID- 10453928 TI - Recovery of the torque-velocity relationship after short exhausting cycling exercise. AB - The effects of fatigue upon the torque-velocity (T-omega) relationship in cycling were studied in 11 subjects. Fatigue was induced by short exhausting exercise, on a cycle ergometer, consisting of 4 all-out sprints without recovery. The linear (T-omega) relationship was determined during each all-out sprint, before, during and after the exhausting exercise. The kinetics of the T-omega relationship had permitted the study of the recovery of optimal torque, optimal velocity and their corresponding maximal power outputs (Pmax), 30 s or 1 min after the short exhausting exercise. Fatigue induced a parallel shift to the left of the T-omega relationship which was partly reversed by a parallel shift to the right during recovery. After 30 s recovery optimal velocity, optimal torque and Pmax were slightly lower than the corresponding values before the exhausting exercise; after 1-min optimal velocity and optimal torque had recovered 99% and 97% of their initial values. These mechanical data suggested that the causes of exhaustion were processes that allowed fast recovery of both optimal velocity and optimal torque. PMID- 10453929 TI - Antigenic characterization of pre- and post-liver transplant hepatitis B surface antigen sequences from patients treated with hepatitis B immune globulin. AB - BACKGROUND/AIMS: The success of treatment with hepatitis B hyperimmune globulin in preventing recurrence of hepatitis B virus infection in patients undergoing orthotopic liver transplantation depends on maintaining levels of anti-HBs sufficient to neutralise hepatitis B virus and also on patient compliance. Breakthrough infections may occur, and these have been associated with the emergence of variants in HBsAg. METHODS: Three patients, two who relapsed and one who had no evidence of hepatitis B virus infection post-orthotopic liver transplantation were studied. Polymerase chain reaction and sequencing of pre- and post-orthotopic liver transplantation samples was followed by antigenic analysis of the in vitro expressed cloned sequences. RESULTS: In two patients who were treated with hyperimmune globulin, amino acid variation in the region of the immunodominant B cell epitopes of HBsAg occurred. Sequencing of clones revealed fluctuating variant sequences over time. One had clinical relapse and immune escape was evident on in vitro antigenic analysis. Patient two lost HBsAg reactivity post-orthotopic liver transplantation. There was loss of an antigenically critical cysteine molecule; sequencing of clones revealed that this was the dominant species. The third patient relapsed when protective levels of anti-HBs were not maintained; HBsAg showed no variation compared to a standard subtype sequence. CONCLUSION: These data provide strong experimental evidence of immune escape. It appears that hyperimmune globulin provides the selection pressure. In these patients, HBsAg negativity does not exclude infection of the transplanted liver. PMID- 10453930 TI - Hematopoietic stem cell transplantation from non-replicative hepatitis B virus carriers is safe. AB - BACKGROUND/AIMS: Hepatitis B virus can cause serious problems in individuals undergoing organ transplantation. The aim of this study was to evaluate the hepatic events among HBs-Ag positive recipients and HBs-Ag negative recipients who received products from hepatitis B virus carriers. METHODS: A total of 151 patients received an allogeneic hematopoietic stem cell transplantation at the Department of Hematology-Oncology, University of Ankara, between June 1989 and June 1998. Among these, eight HBs-Ag positive and four HBs-Ag negative recipients received a product from a hepatitis B virus positive donor. The median follow-up period for these 12 patients was 13.2 months. RESULTS: Three of the eight HBs-Ag positive recipients died (one from hepatic failure); of the remainder, two are HBs-Ag negative, two HBs-Ag positive with normal liver injury tests and one HBs Ag positive with elevated ALT levels. Of the four HBs-Ag negative recipients who received stem cells from a hepatitis B positive donor, two died; none of the patients in this group became HBs-Ag positive after transplantation. CONCLUSION: Hepatitis B virus infection is a common problem in patients being considered for allogeneic hematopoetic stem cell transplantation, especially in areas where hepatitis B virus infection is endemic. We believe that the presence of HBs-Ag positivity is not an absolute contraindication for allogeneic hematopoetic stem cell transplantation unless the hepatitis B virus is in a replication phase. PMID- 10453931 TI - Oral ganciclovir treatment in chronic hepatitis B virus infection: a pilot study. AB - BACKGROUND/AIMS: Ganciclovir is a nucleoside analogue with an excellent safety record; it is effective against cytomegalovirus infection in both its intravenous and its oral form. Intravenous administration of ganciclovir is active against hepatitis B virus but the efficacy of oral ganciclovir is unknown. The aim of this study was to evaluate the tolerability and primary efficacy in reducing HBV DNA levels and liver enzymes of 2 dosing schedules of oral ganciclovir in HBeAg positive and -negative patients with chronic hepatitis B. METHODS: Oral ganciclovir was administered to 15 consecutive patients with active chronic hepatitis B (age 43+/-12 years; 73% males; seven HBeAg-positive and eight negative; no cirrhosis) in a pilot, phase I, open-label study. Before treatment, all patients were screened for 8 weeks to ascertain the persistence of biochemical and virological activity, and then randomized to receive 3 g (eight patients), or 6 g (seven patients) of oral ganciclovir daily for 8 weeks; following therapy, they were closely observed for 8 more weeks. RESULTS: Baseline HBV DNA declined by 99% or 2 log10 at the end of treatment (405.0 vs 3.9 MEq/ml, respectively) and in four patients serum HBV DNA became undetectable by the Monitor assay. Oral ganciclovir suppressed HBV equally well in HBeAg-positive and -negative patients, and the 3- and 6-g daily dose regimens were equally effective. The pre-treatment viral load, however, was a determinant of response, with patients in the lowest quartile of baseline HBV DNA levels responding significantly better than those in the upper quartile (3.0 vs 0.6 log10 respectively, p=0.002). Serum alanine aminotransferase levels became normal or declined in most patients. Oral ganciclovir was very well tolerated. Within 8 weeks after stopping medication, a relapse in serum HBV DNA levels occurred in nine of the 15 patients (60%). CONCLUSIONS: These findings suggest that ganciclovir administered orally at a dose of 3 g can achieve sufficient suppression of HBV replication. This dose, and even higher doses, are well tolerated and could be used as an alternative or in combination with other antivirals for the treatment of chronic hepatitis B. PMID- 10453932 TI - Effects of long-term interferon-alpha treatment on glucose tolerance in patients with chronic hepatitis C. AB - BACKGROUND/AIMS: Interferon-a has been reported to acutely induce insulin resistance and glucose intolerance. The effects of long-term treatment with interferon-a on glucose metabolism remain unclear. METHODS: Thirty-two Japanese patients with chronic hepatitis C were given interferon-a (6x10(6)U/day) daily for 2 weeks and thereafter 3 times weekly up to 6 months. The patients received a 75-g oral glucose tolerance test before the treatment. Fifteen patients also had an intravenous glucose tolerance test for an assessment of insulin sensitivity with Bergman's minimal model. These tests were repeated 3 months after the treatment. RESULTS: Insulin sensitivity was not affected by the treatment (5.7+/ 3.8 vs 5.2+/-3.8 10(-4) x min(-1) x mU(-1) x l , not significant) and a statistically significant but minimum decrease in area under the curve of plasma glucose (1012+/-332 vs 928+/-282 mmol x l(-1) x min, p<0.01) in a 75-g oral glucose tolerance test was noted. Acute insulin response to intravenous glucose tolerance tests (214+/-275 vs 294+/-334 mU x l(-1) x min, p<0.05) increased slightly. CONCLUSION: Contrary to the known acute metabolic effects, interferon-a therapy for 3 months in patients with chronic hepatitis C did not have deleterious effects on insulin sensitivity and glucose tolerance. PMID- 10453933 TI - High prevalance of TT virus infection in Japanese patients with liver diseases and in blood donors. AB - BACKGROUND/AIMS: Although a novel DNA virus, TT virus (TTV), has been isolated from a patient with cryptogenic post-transfusion hepatitis, its pathogenic role remains unclear. To elucidate its prevalence and clinical impact in patients with liver diseases, the presence of TTV DNA was assessed in patients with liver diseases and blood donors (BDs) in Japan using two primer sets, one conventional and the other new and highly sensitive. METHODS: We studied 261 samples, 72 with chronic hepatitis associated hepatitis C virus (HCV-CH), 57 with hepatocellular carcinoma associated HCV (HCV-HCC), 12 with HCC without either HCV or hepatitis B virus (NBNC-HCC), and 120 of BDs. RESULTS: Using two primer sets, TTV DNA was detected in 68 (94.4%), 53 (93.0%), 12 (100%), and 98 (81.7%) HCV-CH, HCV-HCC, NBNC-HCC, and BDs, respectively. The prevalence was not significantly different between HCV-CH and HCV-HCC, or between HCV-HCC and NBNC-HCC. Comparison between patients with and without TTV revealed no significant differences in backgrounds or biochemical findings. Histopathological findings in patients with HCV-CH, and number, maximum diameter, and histological differentiation of HCC also did not demonstrate any relation to TTV infection. TTV strains can be divided into five groups using phylogenetic analysis, but no disease-specific group appears to exist. CONCLUSIONS: Our data suggest that: 1) TTV is very prevalent among patients with liver diseases and even among BDs in Japan, 2) TTV infection does not impact on liver damage with HCV infection, and 3) TTV infection also does not affect the development or progression of HCC. PMID- 10453934 TI - Inhibition of hepatic cell nuclear DNA fragmentation by zinc in carbon tetrachloride-treated rats. AB - BACKGROUND/AIMS: The aims of this study were to ascertain: 1) whether hepatic cell DNA fragmentation is increased in rats with early stages of liver disease induced by carbon tetrachloride; 2) whether the inhibition of DNA cleavage is involved in the hepatoprotective effects of zinc; and 3) if relationships exist between DNA fragmentation and the onset of fibrosis in this experimental model. METHODS: Twenty-one treated rats and 23 controls were divided into two groups to receive either a standard diet or one supplemented with zinc. All the animals were sacrificed 1 week later for histological and biochemical assessments, which included a DNA fragmentation index, hepatic zinc and metallothionein concentrations, fibrosis measured by hepatic hydroxyproline concentration and plasma alanine aminotransferase activity. RESULTS: Hepatic cell DNA fragmentation was increased in rats with early hepatic fibrosis and the increase was independent of hepatocytolysis, as measured by alanine aminotransferase activity. Oral zinc administration inhibited hepatic cell DNA fragmentation in the treated rats and was proportional to the hepatic concentration of the metal. The mechanism of the zinc-mediated decrease in DNA cleavage was related to an increase in the hepatic metallothionein concentration. Hepatic cell DNA fragmentation was related to hydroxyproline concentration. CONCLUSIONS: Our results suggest that apoptosis may be involved in the early transformations occurring in the liver and which can lead to the initiation of cirrhosis. As such, the potential therapeutic use of zinc supplementation would warrant further investigation. PMID- 10453935 TI - Splenectomy attenuates superoxide anion release into the hepatic sinusoids after lipopolysaccharide challenge. AB - BACKGROUND/AIMS: The aim of this study was to determine whether the spleen contributes to superoxide anion release into the hepatic sinusoids and subsequent damage to endothelial cells of the hepatic sinusoids after lipopolysaccharide challenge. METHODS: Rats were given 2 mg/kg body weight lipopolysaccharide. Three hours after the treatment, superoxide anion release into the hepatic sinusoids was examined in a liver perfusion model using the cytochrome C method. Damage to endothelial cells of the hepatic sinusoids was assessed from the purine nucleoside phosphorylase/glutamic-pyruvic transaminase ratio in the liver perfusate. To further characterize the mechanisms behind these changes, these studies were done in rats given superoxide dismutase or an anti-TNFalpha antibody. To study whether the spleen plays a role in the mechanisms, experiments with splenectomized rats were performed. RESULTS: Lipopolysaccharide challenge resulted in superoxide anion release into the hepatic sinusoids and damage to endothelial cells of the hepatic sinusoids. These changes were significantly attenuated by the treatments with superoxide dismutase or an antibody against TNFalpha, as well as by splenectomy. The hepatic macrophage and Kupffer cell populations after lipopolysaccharide challenge were significantly smaller in the rats given splenectomy than in those given a sham operation. There were no significant differences in the neutrophil populations between the two groups. Levels of TNFalpha were significantly lower in the former than the latter, whereas there were no significant differences in levels of Interleukin-8 between the two groups. CONCLUSIONS: Splenectomy reduced the superoxide anion release into the hepatic sinusoids caused by the lipopolysaccharide challenge and subsequent damage to endothelial cells of the hepatic sinusoids. This supports the view that splenectomy has a protective effect in lipopolysaccharide-induced liver injury. PMID- 10453936 TI - Primary biliary cirrhosis shows association with genetic polymorphism of tumour necrosis factor alpha promoter region. AB - BACKGROUND/AIMS: Primary biliary cirrhosis is an autoimmune disease in which increased prevalence in first-degree relatives and an association with HLA DR8 suggest a genetic background. TNFalpha is a mediator of inflammation and immunity, and is implicated in the pathogenesis of primary biliary cirrhosis, ex vivo studies having shown reduced production of TNFalpha by lymphocytes from patients. Our group has previously described a biallelic promoter-region polymorphism of the TNFA gene at position -308, and demonstrated that the rare allele, TNF*2, has increased promoter function compared with the common allele, TNF*1. A further biallelic base change has been described in the TNFA gene at 238. We conducted a case-control study to assess association of these gene polymorphisms with primary biliary cirrhosis. METHODS: Ninety-one patients and 213 controls were genotyped for both TNFA loci using restriction fragment length polymorphism analysis of PCR products. RESULTS: The high production TNFA-308*2 allele was significantly under-represented among subjects with primary biliary cirrhosis (27.5% PBC, 41.6% controls, p=0.02, pc=0.04, OR for carriage of TNF*1/*1 genotype=1.89, CI=1.10-3.32). No association was shown with the TNFA 238 polymorphism. CONCLUSION: Primary biliary cirrhosis is associated with reduced carriage of TNF*2. This is in keeping with a protective role of TNFalpha against the disease. PMID- 10453937 TI - Expression of collagens type I and IV, osteonectin and transforming growth factor beta-1 (TGFbeta1) in biliary atresia and paucity of intrahepatic bile ducts during infancy. AB - BACKGROUND/AIMS: Biliary atresia and paucity of intrahepatic bile ducts are the main causes of neonatal cholestasis leading to hepatic fibrosis. Fibrotic evolution is slow in paucity of bile ducts as compared to the rapid progression to biliary cirrhosis in biliary atresia when cholestasis persists despite hepatoportoenterostomy. Our aim was to compare the expression of collagens type I and IV, alpha-smooth muscle actin, osteonectin and transforming growth factor beta1 in biliary atresia and paucity of bile ducts. METHODS: Liver biopsies were obtained in 12 children with biliary atresia and in five with paucity of bile ducts. Collagens type I and IV, alpha-smooth muscle actin were detected with immunostaining. Collagens type I and IV, osteonectin and transforming growth factor beta1 mRNAs were detected by in situ hybridization. RESULTS: Expression of mRNA and proteins was roughly parallel. In ductular proliferation areas of biliary atresia: (1) the expression of collagens type I and IV and osteonectin was increased, and was localized to periductular myofibroblasts; (2) transforming growth factor beta1 was expressed around biliary ductules, probably in inflammatory cells, and also in biliary cells. Osteonectin expression was also increased in the lobules. In paucity of bile ducts, there was no overexpression of collagens type I and IV and transforming growth factor beta1, except in the only child with marked fibrosis. However, osteonectin expression was enhanced at the periphery of the lobules, even when fibrosis was mild or absent. CONCLUSIONS: These findings suggest that in biliary atresia ductular proliferation areas are the site of a marked production of extracellular matrix proteins in periductular myofibroblasts, probably secondary to transforming growth factor beta1 production by inflammatory cells and by biliary cells. The weak expression of transforming growth factor beta1 could explain the slow progression of fibrosis in paucity of bile ducts. PMID- 10453938 TI - Prognostic factors and long-term effects of ursodeoxycholic acid on liver biochemical parameters in patients with primary biliary cirrhosis. Dutch Multi Centre PBC Study Group. AB - BACKGROUND/AIMS: Serum bilirubin is a prognostic factor in untreated primary biliary cirrhosis (PBC), but this has been less extensively documented for patients treated with UDCA. The aims of this study were to define the effects of UDCA on serum liver tests and to assess prognostic factors in patients on prolonged UDCA treatment. METHODS: Analysis of laboratory parameters obtained before and during treatment with UDCA of 203 PBC patients who were followed for a mean of 48 months. Univariate and multivariate analyses were performed to assess the prognostic value of pre-entry and follow-up variables with respect to treatment failure and survival. RESULTS: Actuarial 5-year incidences of treatment failure and transplantion-free survival were 27 and 79%, respectively. According to the univariate analysis the following variables were significantly associated with prognosis: pre-entry presence of cirrhosis and pre-treatment levels of serum bilirubin and albumin, bilirubin levels during follow-up, the occurrence of biochemical remission and normalisation of serum bilirubin. Multivariate analysis revealed that bilirubin during follow-up was the best predictor. Alkaline phosphatase, aspartate aminotransferase and IgM decreased significantly during the first 6 months of treatment and subsequently remained at this lower level. Serum bilirubin showed the same initial pattern, but a significant increase was observed after 4 years of treatment. CONCLUSIONS: Serum bilirubin in both UDCA treated and untreated patients is the most powerful predictor of prognosis for PBC. The partial therapeutic efficacy of UDCA is illustrated by the finding that serum bilirubin, in contrast to alkaline phosphatase and the transaminases, appears to increase after 4 years of treatment. PMID- 10453939 TI - Ursodeoxycholic acid prevents hepatic cytochrome P450 isozyme reduction in rats with deoxycholic acid-induced liver injury. AB - BACKGROUND/AIMS: Hydrophobic bile acids, such as deoxycholic acid produce cholestatic liver injury. Ursodeoxycholic acid has been shown to be useful in the treatment of cholestatic liver disease. METHODS: In this study, we investigated the effects of deoxycholic acid or ursodeoxycholic acid (1% of diet, for 14 days) and their combination (1% each) on expression of hepatic cytochrome P450 isozymes, their related enzyme activities and mRNA level in rats. RESULTS: Adding 1% deoxycholic acid to chow caused a marked increase in serum total bilirubin (47 fold) and total bile acid (8-fold) concentrations and in alkaline phosphatase (2.5-fold, p<0.01) and alanine aminotransferase activities (23.5-fold, p<0.01). Adding the same dose of ursodeoxycholic acid along with the deoxycholic acid mitigated both the rise in serum total bilirubin and bile acid concentrations and that in alkaline phosphatase and alanine aminotransferase activities, although the use of ursodeoxycholic acid alone did not affect any of the above. Feeding 1% deoxycholic acid caused a decrease (48% of control) in total cytochrome P450 content in hepatic microsomes. Addition of 1% ursodeoxycholic acid along with the 1% deoxycholic acid completely prevented the decrease in total cytochrome P450 content. Feeding ursodeoxycholic acid alone did not affect the total cytochrome P450 content. The expression of cytochrome P450 2B1, 2E1, 3A2, 2C6, 2C11 and 4A1 proteins in hepatic microsomes was decreased by deoxycholic acid (44, 51, 23, 59, 30 and 74% of control, respectively). Likewise, the activities of cytochrome P450 2B1 (pentoxyresorufin O-depentylation), 2E1 (aniline p-hydroxylation) and 3A2 (testosterone 6beta-hydroxylation) isozymes and the 3A2 mRNA levels in liver were decreased by deoxycholic acid. Addition of 1% ursodeoxycholic acid to 1% deoxycholic acid also prevented the decrease in these cytochrome P450 proteins, related enzyme activities and mRNA levels in liver. CONCLUSIONS: These results indicate that, in rats with deoxycholic acid-induced liver injury, ursodeoxycholic acid prevents the decrease in hepatic cytochrome P450 isozymes and suggest that ursodeoxycholic acid is useful for the treatment of liver injury in terms of aiding the normalization of the hepatic drug-metabolizing system. PMID- 10453940 TI - Growth hormone/insulin-like growth factor 1 axis alterations contribute to disturbed protein metabolism in cirrhosis patients after hepatectomy. AB - BACKGROUND/AIM: Liver cirrhosis is accompanied by a fall in whole-body protein turnover and alterations of the growth hormone (GH)/insulin-like growth factor 1 (IGF-1) axis. However, the influence of liver cirrhosis on the GH/IGF-1 axis in the perioperative period, and the contribution of the GH/IGF-1 axis alteration in cirrhosis to postoperative nitrogen metabolism are not known. METHODS: Plasma GH, IGF-1 and IGF binding protein 3 (IGF-BP3) levels were measured sequentially in patients undergoing hepatectomy with or without cirrhosis. Postoperative nitrogen excretion and whole-body protein turnover rate were also determined. RESULTS: Preoperative plasma IGF-1 level showed a negative correlation with indocyanine green retention rate. Cirrhosis patients undergoing hepatectomy had low IGF-1 and IGF-BP3 levels, despite extremely high GH levels in the perioperative period. Perioperative IGF-1 levels were lower in patients with postoperative complications than in those without complications. Postoperative nitrogen excretion was higher and whole-body protein turnover rate was lower in patients with cirrhosis than in those without cirrhosis. The post-operative IGF-1 level showed a positive correlation with whole-body protein turnover rate. CONCLUSIONS: Postoperative hepatic IGF-1 production may be severely disturbed in patients with cirrhosis, and the impaired IGF-1 production contributes to the suppressed postoperative protein metabolism in cirrhosis. It may be appropriate to increase plasma IGF-1 level in patients with cirrhosis to enhance postoperative protein metabolism and improve the postoperative outcome. PMID- 10453941 TI - Development of quinolone-resistant strains of Escherichia coli in stools of patients with cirrhosis undergoing norfloxacin prophylaxis: clinical consequences. AB - BACKGROUND/AIM: Norfloxacin prophylaxis decreases the incidence of bacterial infections in high-risk cirrhotic patients, but may promote the development of quinolone-resistant gram-negative bacteria in stools, and eventually lead to infections due to these bacteria. The aim of the study was to evaluate the prevalence of quinolone-resistant strains of E. coli in stools on admission, and the characteristics of any nosocomial infections. METHODS: Eighty-three consecutively hospitalized cirrhotic patients were included in this prospective study. The presence of quinolone-resistant strains of E. coli in stools on admission, and the characteristics of any nosocomial infections were recorded. RESULTS: Fourteen out of 83 patients (16.8%) showed quinolone-resistant E. coli in stools (Group I), and 69 did not (Group II). Thirteen out of 14 from Group I (92.8%) and 17/69 (24.6) from Group II had received primary or secondary prophylaxis with norfloxacin (p<0.001). During hospitalization, 12/12 (100%) of patients from Group I and 25/66 (37.8%) of patients from Group II underwent norfloxacin prophylaxis. Three bacterial infections in patients from Group I, 3 from Group II patients receiving norfloxacin and 16 from Group II patients not receiving norfloxacin were recorded (p<0.05). No infections due to quinolone resistant E. coli were observed in patients colonized with these bacteria. Treatment with norfloxacin induced the development of quinolone-resistant E. coli in 6/14 (42.8%) patients in a mean time of 18.5+/-9.8 days. CONCLUSIONS: The development of quinolone-resistant strains of E. coli is significantly associated with previous administration of norfloxacin prophylaxis. However, in our series this fact is not associated with an increased incidence of quinolone-resistant E. coli or other gram-negative infections. PMID- 10453942 TI - Chronic liver injury alters basal and stimulated nitric oxide production and 3H thymidine incorporation in cultured sinusoidal endothelial cells from rats. AB - BACKGROUND/AIM: Under pathological conditions the nitric oxide synthase (NOS) mediated nitric oxide production of sinusoidal endothelial cells might be altered. Therefore, studies were performed to evaluate the nitrite formation by cultured sinusoidal endothelial cells from rat livers chronically injured by thioacetamide and the effect of endogenously or exogenously generated nitric oxide on their proliferative activity. METHODS: Basal and stimulated nitrite formation, expression of NOS and DNA synthesis were examined in sinusoidal endothelial cells isolated and cultivated from livers with incipient or advanced chemically-induced cirrhosis. RESULTS: Cultured sinusoidal endothelial cells from injured livers exhibited a reduced basal and an increased lipopolysaccharide stimulated nitrite production when compared with controls. Western blot analysis revealed a markedly reduced protein expression of endothelial NOS (eNOS) and inducible NOS (iNOS) in sinusoidal endothelial cells from both experimental groups when compared with controls. Lipopolysaccharide stimulated iNOS expression in sinusoidal endothelial cells from control livers only marginally, and from those with cirrhosis more strongly. There was no clear correlation between the amount of enzyme and nitrite formation. Cultured sinusoidal endothelial cells from livers with incipient cirrhosis showed a higher proliferative activity than controls. Endogenously-produced nitric oxide inhibited DNA synthesis in all groups in a cGMP-independent way. Exogenously-generated nitric oxide affected DNA synthesis differently in sinusoidal endothelial cells from controls and injured livers. CONCLUSION: The results provide evidence that cultured sinusoidal endothelial cells from controls and livers with incipient or advanced cirrhosis differ with respect to basal and lipopolysaccharide-stimulated nitrite production. The data can be taken as evidence that in sinusoidal endothelial cells from livers chronically injured by thioacetamide, eNOS and iNOS are aberrantly expressed and differently regulated. PMID- 10453943 TI - Investigation of albumin-synthesizing ability in rat cirrhotic liver-derived hepatocytes using primary hepatocyte culture. AB - BACKGROUND/AIMS: In cirrhosis, despite a decrease in the total number of hepatocytes, a normal serum albumin level is maintained during the compensatory stage of the disease in many cases. Therefore, to elucidate the mechanism in hepatocytes related to the regulation of the serum albumin level, the albumin synthesizing ability of individual hepatocytes was investigated in cirrhotic rats. METHODS: Cirrhotic rats were prepared by oral administration of furfural to male Wistar rats for 20 weeks. Albumin-synthesizing abilities of liver and of isolated hepatocyte culture were evaluated by measuring the albumin concentration in blood and culture supernatant. Expressions of albumin mRNA were compared using Northern blotting. Furthermore, transcriptional activity of the albumin gene was measured using the promoter domain of the gene. RESULTS: The total number of hepatocytes in rat cirrhotic liver was significantly decreased compared to that in normal rat liver. However, there were no significant differences in levels of serum albumin or albumin mRNA expression between cirrhotic and normal liver. In primary hepatocyte culture, albumin mRNA expression, the amount of albumin secretion and the albumin promoter activity were clearly enhanced in cirrhotic hepatocytes compared to normal hepatocytes. CONCLUSION: Although the total number of hepatocytes was decreased in the rat cirrhosis models used in this study, the serum albumin level was maintained and albumin-synthesizing ability was enhanced at the transcriptional level in the individual hepatocytes. These results suggest that the maintenance of serum albumin levels in compensated cirrhosis may be due to enhanced albumin synthesis by the hepatocytes. PMID- 10453944 TI - Intraportal administration of glyceryl trinitrate or nitroprusside exerts more systemic than intrahepatic effects in anaesthetised cirrhotic rats. AB - BACKGROUND/AIMS: Increased intrahepatic vascular tone can be pharmacologically manipulated in isolated cirrhotic livers. Intrahepatic endothelial dysfunction may lead to a decreased production of the potent endogenous vasodilator nitric oxide in cirrhotic livers. The aims of the study were to determine whether portal pressure can be lowered in vivo by injecting nitric oxide donors glyceryl trinitrate or nitroprusside directly in the portal vein and whether this is related to a decrease in intrahepatic resistance. METHODS: In anaesthetised CCl4 cirrhotic rats, intraportal doses of glyceryl trinitrate 0.5, 1 or 5 microg/kg/ min or nitroprusside 1, 5 or 10 microg/kg/min did not decrease portal pressure but only arterial pressure. Systemic and splanchnic haemodynamics were measured before and during 15 min intraportal infusion of glyceryl trinitrate 10 microg/kg/min or nitroprusside 20 microg/kg/min. RESULTS: Glyceryl trinitrate decreased portal pressure from 14.0+/-1.1 to 11.8+/-1.4 mm Hg, splanchnic perfusion pressure from 102+/-10 to 74+/-5 mm Hg and portal sinusoidal flow from 2.11+/-0.38 to 1.70+/-0.35 ml/min/g liver (all p<0.05). Nitroprusside did not decrease portal pressure significantly but led to a reduction of the splanchnic perfusion pressure (104+/-9 to 66+/-7 mm Hg) and the portal sinusoidal flow (2.39+/-0.50 to 1.77+/-0.31 ml/min/g liver; all p<0.05). Portal sinusoidal resistance was not altered by either drug. CONCLUSIONS: Intraportal infusion of nitric oxide donors decreased arterial pressure more than portal pressure. Portal sinusoidal resistance remained unaffected, but the liver parenchyma became less perfused with high doses. The systemic effects of nitric oxide donating drugs prevailed. PMID- 10453945 TI - Tumour suppressor protein p53 released by nuclease digestion increases at the onset of rat liver regeneration. AB - BACKGROUND/AIMS: Protein kinase CK2 (CK2) increases when cells are committed to proliferate, as in liver regeneration. This enzyme phosphorylates the tumour suppressor protein p53, whose expression controls the levels of many other cell cycle proteins. The aim of this study was to determine if CK2 was affected by p53. METHODS: Male Sprague-Dawley rats (200-250 g) were subjected to either partial hepatectomy or laparotomy and the levels and subcellular distribution of p53 were studied, following the approach used earlier for CK2. The levels of both proteins were also studied in the human cell lines HL-60 (devoid of p53) and HepG2 (with normal p53 levels) and in fibroblasts from transgenic p53-deficient mice (p53-/-) or homozygous for wild-type p53 (p53+/+). Computer-assisted search was used to detect p53 consensus sequences in genes for CK2 subunits Binding of p53 protein to some of these sequences was assayed by electrophoretic mobility shift assay. RESULTS: Rat liver p53 protein was present mainly in the fraction extracted from intact nuclei by nucleases (S1) and showed a transient increase at 6 h post partial hepatectomy, as observed previously with nuclear CK2. The human CK2a gene presents the consensus sequence for trans-activation by p53 and specific binding of p53 protein to some of these sequences was detected in vitro. Total CK2a was higher in HepG2 than in HL-60 cells but total CK2 and its cytosolic/ nuclear distribution was similar in mice (p53+/+) fibroblasts and (p53 /-) fibroblasts. CONCLUSIONS: p53 is present in the nuclease-extracted S1 fraction from liver cells, as described for CK2, and undergoes similar changes at the beginning of rat liver regeneration. However, the data on cultured cells suggest that the expression of CK2 and its subcellular localization are p53 independent events. PMID- 10453946 TI - Overexpression of Bcl-2 protects human hepatoma cells from Fas-antibody-mediated apoptosis. AB - BACKGROUND/AIMS: Fas is a cell surface antigen, that triggers apoptosis upon specific ligand or antibody binding. The proto-oncogene bcl-2 prevents apoptosis induced by various treatments. The aim of our study was to evaluate whether Bcl-2 protects hepatoma cells from Fas-mediated apoptosis. METHODS: Two human cell lines, HCC-T and HepG2 were used. Expression of Fas antigen and Bcl-2 was detected by flow cytometry and Western blotting. Cell viability and apoptotic change were examined after anti-Fas- and antisense oligodeoxynucleotide treatments. Apoptotic cells were detected by nick-end labelling and the TUNEL method. To test if Bcl-2 expression can protect HepG2 cells from Fas-mediated apoptosis, the cells were transduced using retroviral vector, LZBC, designed to coexpress E. coli beta-galactosidase and human Bcl-2. To further confirm the protective effect of Bcl-2 expression against Fas-mediated apoptosis in HepG2, Bcl-2 expressing plasmid vector was produced and a cell line stably expressing Bcl-2 was cloned. RESULTS: Western blot analysis showed constitutive Bcl-2 expression in HCC-T cells, but not in HepG2 cells. HCC-T was resistant to apoptosis after treatment with an agonist anti-Fas antibody (1 microg/ml for 3 days), whereas 33% of the HepG2 cells were killed by this treatment. Inhibition of Bcl-2 expression by transfection of antisense oligodeoxynucleotides caused spontaneous apoptosis in HCC-T, but not in HepG2 cells, suggesting that Bcl-2 is essential for survival of HCC-T cells, whereas other proteins may substitute for it in HepG2 cells. Following LZBC infection, 10% HepG2 cells were beta galactosidase-positive by X-gal staining and Bcl-2-positive. In cells surviving after anti-Fas treatment, the proportion of beta-galactosidase-positive cells increased to 50% and the beta-galactosidase activity increased 6-fold, indicating that Bcl-2 expression protected the cells from Fas-mediated apoptosis. In the cloned HepG2 cells stably expressing Bcl-2, the extent of Fas-mediated apoptosis was inversely related to the level of Bcl-2 expression. CONCLUSION: Bcl-2 confers protection to human hepatoma cells against Fas-mediated apoptosis, and is essential for survival of some, but not all, hepatoma cells. PMID- 10453947 TI - Dendritic cells with immature phenotype and defective function in the peripheral blood from patients with hepatocellular carcinoma. AB - BACKGROUND/AIMS: Defects and dysfunctions in antigen-presenting dendritic cells have been shown during carcinogenesis. The phenotype and function of dendritic cells have been studied in patients with hepatocellular carcinoma to explore the possibility of dendritic cell-based immune therapy in hepatocellular carcinoma. METHODS: The stimulatory capacity of dendritic cells in allogenic mixed leukocytes reaction, the expression of surface makers on dendritic cells, the production of cytokines and nitric oxide by dendritic cells and the levels of maturation of dendritic cells from 17 patients with hepatocellular carcinoma, 10 patients with liver cirrhosis and 10 normal controls were compared. RESULT: Dendritic cells from hepatocellular carcinoma had significantly lower capacity to stimulate allogenic T cells in allogenic mixed leukocytes reaction compared with dendritic cells from liver cirrhosis and normal controls (p<0.05). Dendritic cells from hepatocellular carcinoma expressed significantly lower levels of HLA DR and induced decreased amounts of interleukin-12 compared with dendritic cells from normal controls (p<0.05). On the other hand, dendritic cells from hepatocellular carcinoma produced significantly higher levels of nitric oxide and tumor necrosis factor-a compared with dendritic cells from liver cirrhosis and normal controls (p<0.05). The uptake of fluorescein isothiocyanate-labeled dextran revealed an immature phenotype of dendritic cells from hepatocellular carcinoma compared with dendritic cells from liver cirrhosis (p<0.05). CONCLUSION: The results of this study on the function of dendritic cells in hepatocellular carcinoma, and the prevalence of immature dendritic cells in hepatocellular carcinoma in the microenvironment of high levels of inflammatory cytokines indicate a specific defect of dendritic cell maturation during hepatocarcinogenesis. These data show that induction of dendritic cell maturation might be an approach to dendritic cell-based immune therapy during hepatocellular carcinoma. PMID- 10453948 TI - Presence of alpha-fetoprotein mRNA in blood correlates with outcome in patients with hepatocellular carcinoma. AB - BACKGROUND/AIMS: Since hematogenous spread of tumor cells may adversely affect the prognosis of patients with hepatocellular carcinoma, we prospectively analyzed whether the presence of alpha-fetoprotein (AFP) messenger RNA (mRNA) in blood, used as a marker of circulating hepatocellular carcinoma cells, correlates with outcome. METHODS: Eighty-eight patients were enrolled between December 1993 and August 1995, and 81 were followed until the end of 1997. All patients were treated with percutaneous ethanol injection therapy and/or transarterial embolization during follow-up. The status of AFP mRNA in blood was serially determined. Cumulative metastasis-free survival and overall survival were analyzed in relation to AFP mRNA and other clinical and laboratory variables. RESULTS: Among 81 patients followed, 54 were positive for AFP mRNA at entry and 27 were negative. Extrahepatic metastasis developed more frequently among the AFP mRNA-positive patients (13 of 54) than among the AFP mRNA-negative patients (2 of 27) (p=0.0296). After treatment, AFP mRNA became negative in 24 of 54 patients (44%). Cumulative metastasis-free survival and overall survival were significantly better in the 24 patients whose AFP mRNA became negative after treatment than in the 30 patients with persistently positive AFP mRNA (p= 0.0001 and p<0.0001, respectively). CONCLUSIONS: The presence or absence of AFP mRNA in blood is a predictor of outcome in patients with hepatocellular carcinoma. PMID- 10453950 TI - MRI guidance of infra-red laser liver tumour ablations, utilising an open MRI configuration system: technique and early progress. AB - BACKGROUND/AIMS: Primary and secondary liver tumours are a common clinical problem, with a poor prognosis in most cases. Surgical resection offers the best outcome, but is only appropriate for the minority. Thermal ablation techniques have been described, but the lack of an optimal means of monitoring has limited their use. We undertook a pilot study to assess the feasibility and safety of an integrated MR-guided laser thermoablation technique under local anaesthesia using a real-time colourisation thermal monitoring technique in a newly developed open MR scanner. METHODS: Liver tumours were punctured after the administration of intravenous Mangafodipir trisodium (MnDPDP) using real-time MR image guidance under local or general anaesthesia, and treated using a water-cooled interstitial fibre and a Nd-YAG laser source. Twenty-seven procedures were performed in 12 patients. Therapy was monitored using a real-time MR colourisation sequence. Thermoablation was followed by a colour change in a region of interest. RESULTS: Thermal lesions of mean size 3 cm in diameter were produced with a maximum size of 5 cm. Eight out of 12 patients were discharged the next day with few significant complications. Repeat procedures have been performed in seven of 12 patients. Two patients with lesions of 3 cm diameter have had complete tumour ablation with only one procedure. CONCLUSION: Percutaneous laser thermoablation for liver tumours performed as an integrated one-step technique in an open configuration MR scanner is described. It can be safely performed under local anaesthesia in the majority of patients, with few side effects. MR control shows the site and size of the evolving thermal lesions, allowing appropriate action to be taken in terms of further burns, time of application and power applied. PMID- 10453949 TI - Effect of transcatheter arterial chemoembolization on kidney hemodynamics and function in patients with cirrhosis and hepatocellular carcinoma. AB - BACKGROUND/AIMS: Transcatheter arterial chemoembolization (TACE) may have deleterious effect on the kidney in patients with cirrhosis and hepatocellular carcinoma. The aim of the study was to test this hypothesis. METHODS: Twenty-four patients with cirrhosis and hepatocellular carcinomas were included. They consisted of 16 patients undergoing a single TACE and eight patients undergoing diagnostic angiography. Doppler ultrasonography was used to measure hepatic artery pulsatility index (HA-PI) and renal artery pulsatility index (RA-PI) before and 1 day and 10 days after the procedure. Similarly, kidney function was assessed by measuring creatinine clearance. In addition, plasma renin activity, noradrenaline, and endothelin-1 were also measured. RESULTS: In patients receiving diagnostic angiography, no significant changes in HA-PI were observed after the procedure. In contrast, HA-PI increased significantly 1 day after the procedure (19%, p<0.01) in patients undergoing TACE, although it returned to baseline value 10 days after the procedure. In patients undergoing diagnostic angiography, no significant changes in RA-PI were observed after the procedure. Similarly, no detectable changes in RA-PI were noted in patients undergoing TACE. A transient small reduction in creatinine clearance was noted after the procedure in patients undergoing diagnostic angiography (-12%, p<0.05) and in those undergoing TACE (-11%, p<0.05). However, the effect was similar in the two groups (two-way ANOVA, p=0.72). No significant changes in plasma renin activity, noradrenaline, and endothelin-1 were observed after either diagnostic angiography or TACE. CONCLUSIONS: These results suggest that TACE per se has no deleterious effect on the kidney hemodynamics and function in patients with cirrhosis and hepatocellular carcinoma. PMID- 10453952 TI - Pulmonary hypertension following hepatopulmonary syndrome in a patient with cirrhosis. AB - We report the case of a patient with liver cirrhosis who successively developed hepatopulmonary syndrome and portopulmonary hypertension. Initially, the patient presented with severe dyspnea and hypoxemia at rest. Technetium-99 macroaggregated albumin lung perfusion scan demonstrated right-to-left shunt, and hemodynamic study revealed a hyperdynamic state with low pulmonary vascular resistance, thus confirming the diagnosis of hepatopulmonary syndrome. More than 2 years after the onset of pulmonary symptoms, a marked improvement in dyspnea and gas exchange was observed. Lung perfusion scan did not disclose any right-to left shunt and right-sided heart catheterization showed evidence of severe pulmonary hypertension. We conclude that hepatopulmonary syndrome and portopulmonary hypertension are not mutually exclusive. We hypothesize that, by reversing the phenomenon of intrapulmonary vasodilatation, the development of portopulmonary hypertension interfered with each of the potential causes of hypoxemia in hepatopulmonary syndrome (ventilation-perfusion inequalities, intrapulmonary shunting, oxygen diffusion limitation) and, as a result, led to a correction of hypoxemia. PMID- 10453951 TI - Direct hyperplasia does not enhance the kinetics of liver repopulation in a new model of hepatocyte transplantation in the rat. AB - BACKGROUND/AIMS: We have recently developed a new model of extensive liver repopulation by transplanted hepatocytes following exposure to pyrrolizidine alkaloids. In the present study, the effect of 2/3 partial hepatectomy (PH) and that of a potent direct liver mitogen, lead nitrate, were compared in their ability to modulate the kinetics of liver repopulation. METHODS: Fischer 344 rats deficient in enzymatic activity for dipeptidyl-peptidase IV (DPPIV-) were used as cell transplantation recipients. They were given 2 doses of the pyrrolizidine alkaloid retrorsine (30 mg/kg, i.p.), 2 weeks apart, followed 2 weeks later by transplantation of 2 x 10(6) hepatocytes (via the portal vein), freshly isolated from a normal congeneic DPPIV+ donor. PH was carried out or a single injection of lead nitrate (100 micromol/kg, i.v.) was administered 2 weeks post transplantation. Liver samples obtained at different time points post-treatment were processed histochemically for DPPIV activity. RESULTS: The percent of liver sections occupied by DPPIV+ hepatocytes was <1% at the time of PH or lead nitrate administration. In animals which underwent PH, it increased to 33.4+/-5.7% at 2 weeks and to 55.6+/-8.5% at 1 month. However, in animals receiving lead nitrate, these percentages were only 3.3+/-1.3% at 2 weeks and 16.2+/-3.9% at 1 month. Repeated injections of lead nitrate had no additional effect. Further experiments indicated that an acute mitogenic response to lead nitrate was present in transplanted cells, while resident hepatocytes were inhibited by retrorsine. CONCLUSIONS: These results indicate that direct mitogenic signals (such as those induced by lead nitrate), and compensatory signals (such as those elicited by PH), are not equally effective on kinetics of liver repopulation in this system. The possible reasons for these differential effects are discussed. PMID- 10453953 TI - Images in Hepatology. Liver abscesses with bilioportal fistula. PMID- 10453954 TI - TNF gene polymorphisms in primary biliary cirrhosis: a critical appraisal. PMID- 10453956 TI - Genetic basis of progressive familial intrahepatic cholestasis. PMID- 10453955 TI - Extrahepatic manifestations of HCV infection: facts and controversies. PMID- 10453957 TI - HCV NS5A 2209-2248 amino acid substitutions affect serum 2,5AS activities. PMID- 10453958 TI - Serum level of interleukin-4 is a possible marker of therapeutic response to interferon treatment in patients with chronic hepatitis C. PMID- 10453960 TI - Adeno-associated virus as a gene delivery vector for liver cells. PMID- 10453959 TI - A controlled trial of gemfibrozil in the treatment of patients with nonalcoholic steatohepatitis. PMID- 10453961 TI - Expression of p21 and p27 in hepatoma cell lines with different p53 gene profile. PMID- 10453962 TI - Pharmacy continuing education available on the Internet. AB - Access to interesting and pertinent continuing education material would be expected to encourage pharmacists to pursue postgraduate professional development. The Internet is one means of providing accessible and timely instructional material. Twenty-nine Internet sources of continuing education available for pharmacists were identified. Criteria for inclusion included free educational credit and accreditation by a recognized body. Evaluation criteria were currency of the offerings, author and sponsor identification, software requirements in addition to a browser, and methods for submitting examination answers and receiving certificates of credit. PMID- 10453963 TI - Correlation of activated clotting time and activated partial thromboplastin time to plasma heparin concentration. AB - STUDY OBJECTIVE: To determine the correlation between activated clotting time (ACT) or activated partial thromboplastin time (aPTT) and plasma heparin concentration. DESIGN: Two-phase prospective study. SETTING: University affiliated community hospital. PATIENTS: Thirty patients receiving continuous infusion intravenous heparin. INTERVENTIONS: Measurement of ACT, aPTT and plasma heparin concentrations. MEASUREMENTS AND MAIN RESULTS: Linear and log linear correlations were determined between clotting time tests and heparin concentrations. Linear correlations yielded r values of 0.58 for ACT (p=0.008) and 0.89 for aPTT (p=0.0001). Log linear correlations yielded r values of 0.60 for ACT (p=0.005) and 0.88 for aPTT (p=0.0001). A decision analysis was performed to determine possible consequences of dosage adjustments based on either test in relationship to the decision based on plasma heparin concentration. The decision analysis based on ACT disagreed with corresponding decisions based on plasma heparin concentration in 15 of 30 patients; 13 disagreements may have increased the risk of bleeding, and the other 2 may have increased the risk of thrombosis. Decisions based on aPTT disagreed with corresponding decisions based on plasma heparin concentration in 13 of 30 patients; 2 disagreements may have increased the risk of bleeding, and the other 11 may have increased the risk of thrombosis. CONCLUSION: There are significant statistical linear and log linear correlations between both clotting time tests and plasma heparin concentrations, with aPTT showing stronger correlation than ACT. However, decisions regarding heparin therapy based on ACT may increase a patient's risk of bleeding, whereas decisions based on aPTT may increase the risk of thrombus progression or rethrombosis. PMID- 10453964 TI - Abacavir: absolute bioavailability, bioequivalence of three oral formulations, and effect of food. AB - STUDY OBJECTIVES: Study A: to determine the absolute bioavailability of a single 300-mg abacavir hemisulfate tablet. Study B: to determine the bioequivalence of two oral abacavir formulations (300-mg hemisulfate tablet, 100-mg succinate caplet), the effect of food on the bioavailability of the 300-mg hemisulfate tablet, and the bioavailability of the hemisulfate tablet relative to the hemisulfate solution. DESIGN: Phase I, randomized, open-label, balanced two- (study A) and three- or four-period (study B), crossover studies. SETTING: Two clinical research centers. SUBJECTS: Six men infected with the human immunodeficiency virus (HIV), aged 27-39 years (study A), and 18 HIV-infected men and women, aged 21-50 years (study B). INTERVENTIONS: In study A, all subjects received a single, oral 300-mg tablet of abacavir hemisulfate or a single, intravenous infusion of abacavir hemisulfate 150 mg over 60 minutes. In study B, all subjects received each of three single-dose treatments: three 100-mg abacavir succinate caplets in a fasted state, one 300-mg abacavir hemisulfate tablet in a fasted state, and one 300-mg abacavir hemisulfate tablet with a high-fat breakfast. Twelve subjects in study B also received a fourth treatment of abacavir hemisulfate 300 mg as an oral solution in a fasted state. Plasma samples collected for 24 hours (study A) or 12 hours (study B), and urine samples collected for 12 hours (study A) were analyzed by validated high-performance liquid chromatographic methods. MEASUREMENTS AND MAIN RESULTS: Abacavir pharmacokinetic parameters were calculated using standard, noncompartmental methods. In study A, the geometric least square (GLS) mean absolute bioavailability of oral abacavir was 83% (range 65-107%). In study B, the hemisulfate tablet was bioequivalent to the succinate caplet, but its time to maximum concentration (Tmax) occurred 30 minutes earlier. Administration of the abacavir hemisulfate tablet with food had no effect on area under the curve from time zero to infinity (AUC0-infinity), decreased maximum concentration (Cmax) by 26%, and delayed Tmax by 38 minutes. The relative bioavailability (GLS mean AUC0 infinity ratio) of the 300-mg abacavir hemisulfate tablet to solution was 101%, Cmax was 11% lower, and Tmax was unchanged. The most common drug-related adverse events associated with abacavir were nausea, vomiting, abdominal pain, and headache, all of which were mild. CONCLUSION: Based on our results, abacavir is safe and well tolerated and can be administered with or without meals. PMID- 10453965 TI - Comparison of iothalamate clearance methods for measuring GFR. AB - STUDY OBJECTIVE: To evaluate the bias and precision of three methods of measuring glomerular filtration rate (GFR) relative to a standard method. DESIGN: Prospective, outpatient study. SETTING: University-affiliated general clinical research center. PATIENTS: Twenty-six patients with various degrees of renal function (GFR range 25-151 ml/min/1.73 m2). INTERVENTIONS: Each patient received iothalamate twice during the study visit, first as a bolus injection and then as a priming dose followed by a constant-rate infusion for 2.5 hours. MEASUREMENTS AND MAIN RESULTS: Plasma (ClpIVB) and renal clearances (ClrIVB) after bolus injection and plasma clearance during constant-rate infusion (ClpINF) were compared with standard renal clearance during constant-rate infusion (ClrINF). All three measures were highly correlated with ClrINF (r>0.90, p<0.001). The mean ClrIVB was not significantly different from ClrINF (106.3+/-30.4 vs 104.2+/-28.5 ml/min/1.73 m2) and provided a precise (8.8%, 95% CI 6.5-11.1%) and unbiased measure of GFR. Both ClpIVB and ClpINF were positively biased; values exceeded ClrINF by 11.8+/-11.1 (p=0.0001) and 10.5+/-12.5 ml/min/1.73 m2 (p=0.0003), respectively. Use of a nonrenal correction factor of 9.8 and 10.5 ml/min/1.73 m2 for infusion and bolus plasma clearance values, respectively, eliminated bias and improved the precision of these methods. CONCLUSIONS: Iothalamate renal clearance after bolus injection is a simple, accurate, and precise measurement of GFR and may be a useful alternative to the standard infusion method in clinical investigations. The corrected plasma clearance provides a simple index of GFR for clinical practice. PMID- 10453966 TI - Testosterone and andropause: the feasibility of testosterone replacement therapy in elderly men. AB - Andropause, a syndrome in aging men, consists of physical, sexual, and psychologic symptoms that include weakness, fatigue, reduced muscle and bone mass, impaired hematopoiesis, oligospermia, sexual dysfunction, depression, anxiety, irritability, insomnia, memory impairment, and reduced cognitive function. Free testosterone levels begin to decline at a rate of 1% per year after age 40 years. It is estimated that 20% of men aged 60-80 years have levels below the lower limit of normal. Although the causal relationship between declining testosterone levels and development of andropause symptoms is not firmly established, administration of testosterone to this population resulted in improvements in many areas. Most studies to date focused on physical benefits of testosterone replacement and failed to assess psychologic symptoms rigorously. Preliminary data suggest that therapy may benefit elderly men with new-onset depression. Testosterone administration is not without problems, the most worrisome being the potential for increased prostate cancer risk. Despite this concern, a limited number of studies administered the hormone weekly for up to 2 years, with only mild increases in prostate-specific antigen over control values. Currently, insufficient evidence, primarily regarding psychologic safety and efficacy, exists to warrant general administration of testosterone to elderly hypogonadal men. Further clinical investigations of this therapy in men with low testosterone levels and andropause symptoms are justified and necessary. PMID- 10453967 TI - Migraine: a comprehensive review of new treatment options. AB - Headaches are among the most common complaints reported to health care professionals and are classified by the International Headache Society as migraine, tension-type, or cluster, with additional subtypes. Classification and etiology of headache should be determined after thorough review of the patient's history. Once diagnosed, migraine can be treated by preventive or abortive measures. Recent developments add new options, including availability of drugs for intranasal administration (sumatriptan, dihydroergotamine) and 5-HT1B/1D agonists (rizatriptan, zolmitriptan, naratriptan, eletriptan). Although placebo controlled trials are available, few comparative clinical trials of these agents have been conducted; however, important pharmacologic, pharmacokinetic, and clinical differences exist among the drugs. PMID- 10453968 TI - Beta-adrenergic-blocking agents in bronchospastic diseases: a therapeutic dilemma. AB - Cardioselective beta-blockers should be administered starting with a low dosage under direct medical observation. Bronchodilators should be readily available or may be coadministered. Because of several advantages, agents such as metoprolol, atenolol, and, in some cases, esmolol should be the first agents considered. In contrast to noncardioselective agents, if bronchospasm occurs, the effect of cardioselective agents is believed to be easier to reverse. Clinicians should avoid noncardioselective beta-blockers in asthmatics, even in small doses, such as those administered as eye drops. For asthmatic patients who are intolerant to noncardioselective beta-blockers, switching to a cardioselective beta-blocker might be a safe alternative. The significance of beta2-blockade usually varies with the patient's ventilatory condition, with more serious consequences being anticipated in patients with more severe asthma. PMID- 10453969 TI - Intracellular phosphorylation of zidovudine in an in vitro hollow fiber model. AB - Combination antiretroviral therapy that includes zidovudine has proved much better in treating human immunodeficiency virus infection than monotherapy Diminished responses to zidovudine, especially when it was given alone, was likely due to factors including interpatient variability in pharmacokinetics, nonadherence, emergence of resistant mutants, and reduced cellular enzymatic processes to phosphorylate the drug. This study evaluated the intracellular metabolism of zidovudine up to 6 weeks using a hollow fiber cellular model system that simulated exposure of cells to steady-state concentrations achieved in humans. The CEM-T4 lymphocytes were exposed to simulated 200-, 600-, and 1200-mg daily doses of zidovudine. Samples were analyzed for monophosphate, diphosphate, and triphosphate metabolites of zidovudine by high-performance liquid chromatography separation and radiochemical detection. The monophosphate metabolite increased as simulated doses increased, but no corresponding increases in the active triphosphate metabolite occurred. In addition, intracellular metabolism of zidovudine did not change after exposure for 6 weeks. These results suggest that the active triphosphate metabolite of zidovudine does not change much when doses are increased or when exposed for at least 6 weeks. Hollow fiber models may be used effectively to investigate intracellular metabolism of antiviral agents and for some duration of time. PMID- 10453970 TI - Diurnal variation in plasma norepinephrine in patients with heart failure. AB - Diurnal variation in plasma norepinephrine (PNE) levels is well documented in healthy individuals but not in patients with heart failure. Therefore, we attempted to determine variations in PNE levels over 24 hours, measured hourly, in six patients with an ejection fraction below 40% and a history of heart failure of longer than 3 months. Three controls without a history of heart failure also were evaluated. Both patients and controls had diurnal variations in PNE, with highest levels occurring during the day and lowest at night. When data in patients were evaluated by 6-hour time intervals the mean value for 6:00 A.M. 12:00 noon was approximately twice as high as 12:00 midnight-6:00 A.M. (689+/-329 vs 338+/-166 pg/ml, p<0.05, respectively). Patients also had significant peak to trough variation in PNE levels compared with controls (959+/-396 vs 386+/-84 pg/ml, p<0.02, respectively). These results suggest that significant intrapatient variations in PNE occur over 24 hours in patients with heart failure. These variations may have to be accounted for when evaluating and treating patients with heart failure. PMID- 10453971 TI - Successful rechallenge with etoposide phosphate after an acute hypersensitivity reaction to etoposide. AB - Hypersensitivity reactions to etoposide are reported infrequently and consist of hypotension, hypertension, flushing, diaphoresis, dyspnea, bronchospasm, and loss of consciousness. A 23-year-old woman experienced acute bronchospasm, tachycardia, hypoxia, and moderate hypertension minutes after an infusion of etoposide was begun. Symptoms resolved within an hour after administration of intravenous fluids, methylprednisolone, diphenhydramine, and oxygen. Subsequently, the patient was given etoposide phosphate without incident. To our knowledge, this is the first report of successful rechallenge with etoposide phosphate after an acute hypersensitivity reaction to etoposide. PMID- 10453972 TI - Vinorelbine tartrate-induced pulmonary edema confirmed on rechallenge. AB - A 67-year-old woman with metastatic breast cancer experienced sudden and profound pulmonary edema within 45 minutes after completion of intravenous administration of vinorelbine tartrate on two occasions. Both times the drug was discontinued and the patient was treated aggressively with oxygen, intravenous furosemide, and a vasodilator. The patient suffered no lasting medical complications due to the reaction. Until clear documentation and the mechanism for occurrence of this reaction are known, patients receiving vinorelbine should be monitored closely, particularly in the first few hours after intravenous administration. PMID- 10453973 TI - Phaeohyphomycosis from Exphiala jeanselmei with concomitant Nocardia asteroides infection in a renal transplant recipient: case report and review of the literature. AB - A 59-year-old black man who received a cadaveric renal transplant 15 months earlier developed subcutaneous nodules on his right upper extremity that were identified as phaeohyphomycosis caused by Exophiala jeanselmei. The man was admitted 4 weeks later with a swollen left arm and had Nocardia asteroides in this area and in the apex of his left lung. He was treated with surgical excision, and itraconazole, imipenem-cilastatin, and trimethoprim sulfamethoxazole. With the potential presence of more than one microorganism in an immunocompromised patient, it is important to identify and differentiate them correctly to direct appropriate therapy. PMID- 10453974 TI - Distant and delayed mitomycin C extravasation. AB - Mitomycin C is a vesicant chemotherapeutic agent used to treat solid tumors. Its ability to cause delayed and remote tissue injury after intravenous administration is reported in the literature. Two cases of delayed and distant mitomycin C extravasation injury occurred in our institution. In both patients, no evidence of acute extravasation was visible during or immediately after administration. Within 48 hours, one patient reported erythema, burning, and pain in the hand contralateral to the administration site. The second patient developed three distinct ulcerated lesions on her forearm within 6 weeks of receiving the agent. The lesions, located at sites of venipunctures, occurred 12 15 cm proximal to the site of mitomycin infusion. Because the drug has the potential to cause such unusual and unanticipated tissue injuries, health care professionals and patients should be aware of this hazard. PMID- 10453975 TI - Delayed-onset ticlopidine-induced cholestatic jaundice. AB - An 86-year-old man experienced a rash approximately 2 weeks after starting ticlopidine therapy, necessitating discontinuation of the drug. About 1 month later, despite discontinuation, he developed jaundice and liver test abnormalities. These resolved gradually over the next few months. Based on case reports and the drug's pharmacokinetic profile, a high index of suspicion for ticlopidine-induced jaundice is prudent in patients with recent exposure to the agent who have evidence of liver damage. PMID- 10453976 TI - Factors to consider before discontinuing heparin in patients who develop thrombocytopenia. PMID- 10453977 TI - Mitochondria, oxidative stress, and antioxidant defences. AB - Mitochondria are strongly involved in production of reactive oxygen species, considered today as the main pathogenic agent of many diseases. A vicious circle of oxidative stress and damage to cellular structures can lead to either cell death by apoptosis or to a cellular energetic decline and ageing. The early involvement of mitochondria in apoptosis includes expression of pro-apoptotic factors, release of cytochrome c from the inter-membrane space and opening of the permeability transition pore: cytochrome c release appears to precede pore opening. The mitochondrial theory of ageing considers somatic mutations (deletions) of mitochondrial DNA induced by oxygen radicals as the primary cause of energy decline; experimentally, Complex I appears to be mostly affected. We have developed the Pasteur effect (enhancement of lactate production by mitochondrial inhibition) as a bio-marker of mitochondrial bioenergetics in human platelets, and found it to be decreased in aged individuals. Cells counteract oxidative stress by antioxidants; among lipophilic antioxidants coenzyme Q is the only one of endogenous biosynthesis; exogenous coenzyme Q, however, may protect cells from oxidative stress in vivo. PMID- 10453978 TI - Antioxidant activity: what do we measure? AB - Inhibition of oxidation of 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) by free radicals generated by decomposition of 2,2'-azobis(2-amidopropane) (ABAP) by antioxidants and biological material was studied. A correlation was found between the ability of various substances to delay the onset of ABTS oxidation and their rapid reduction of the ABTS+* cation radical, and between the ability to reduce the maximal rate of ABTS oxidation and slow reduction of ABTS+*. The length of the lag period of ABTS oxidation was found to be independent of ABTS concentration. Similar decrease of peroxynitrite-induced ABTS+* formation by antioxidants was observed when the antioxidants were added before and after peroxynitrite. All these findings indicate that the main effect of antioxidants in this system is reduction of ABTS+* and not prevention of its formation. Reduction of oxidation products rather than inhibition of their formation may be the predominant mode of action of antioxidants in various assays of antioxidant activity. PMID- 10453979 TI - Decomposition of lipid hydroperoxides enhances the uptake of low density lipoprotein by macrophages. AB - This study examined the roles of low-density lipoprotein (LDL) lipid oxidation and peroxide breakdown in its conversion to a form rapidly taken up by mouse peritoneal macrophages. Oxidation of the LDL without decomposition of the hydroperoxide groups was performed by exposure to gamma radiation in air saturated solutions. Virtually complete decomposition of the hydroperoxides was achieved by treatment of the irradiated LDL with Cu2+ under strictly anaerobic conditions. No uncontrolled LDL uptake by macrophages occurred when the lipoprotein contained less than 150 hydroperoxide groups per particle. More extensively oxidized LDL was taken up and degraded by mouse macrophages significantly faster than the native lipoprotein. The uptake was greatly enhanced by treatment of the oxidized LDL with Cu2+. A significant proportion of the LDL containing intact or copper-decomposed LDL hydroperoxide groups accumulated within the macrophages without further degradation. Treatment of the radiation oxidized LDL with Cu2+ was accompanied by aggregation of the particles. Competition studies showed that the oxidized LDL was taken up by macrophages via both the LDL and the scavenger receptors, whereas the copper-treated lipoprotein entered the cells only by the scavenger pathway. Phagocytosis also played an important role in the metabolism of all forms of the extensively modified LDL. Our results suggest that minimally-oxidized LDL is not recognized by the macrophage scavenger receptors unless the lipid hydroperoxide groups are decomposed to products able to derivatize the apo B protein. PMID- 10453980 TI - Oxidative damage to plant DNA in relation to growth conditions. AB - In this study we investigated the level of 8-oxo-2'-deoxyguanosine (8-oxodG) in DNA of Cardamine pratensis plants subjected to different growth conditions trying to answer the question whether factors like light and water accessibility or low temperature may have an impact on the total DNA oxidative damage. The level of this modified nucleoside was determined using HPLC coupled to UV absorbance and electrochemical detection (HPLC-UV-EC). We did not observe any statistically significant differences in 8-oxodG level between DNA of etiolated and light exposed plants as well as between DNA of regularly watered and drought-subjected plants. In contrast, we have shown that chilling (1 degree C for 28 h) brings about the increase of 8-oxodG level in DNA. PMID- 10453981 TI - Differential regulation of signaling pathways for insulin and insulin-like growth factor I. AB - The insulin receptor (IR) and the insulin-like growth factor receptor I (IGF-IR) have different functions in cell growth, apoptosis, differentation, and transformation. Although some of these differences may be explained by the relative level of receptor expression and receptor structure (alpha and beta subunits), they may also be attributed to differences in intracellular signals generated by insulin and IGF-I. The presence of hybrid receptors (IR alphabeta subunits and IGF-IR alphabeta subunits) making up the heterotetramers has added a new dimension to our understanding of the functional roles of these receptors. However, to date the results of efforts to understand the differences between these two closely related receptors have indicated mostly similarities. For example, both receptors utilize IRS-1/IRS-2 and Shc as immediate downstream adaptors, leading to activation of the Ras, Raf, ERK kinases and PI-3 kinase pathways. We have used the yeast two hybrid system to identify proteins which bind to the activated IGF-IR but not to the IR. The cytoplasmic domain of the IGF IR was used to screen a human fetal brain library and two isoforms of the 14-3-3 family were identified. 14-3-3 proteins are a highly conserved family of proteins which have recently been shown to interact with other components of the mitogenic and apoptotic signaling pathways, including Raf, BAD, Bcr/Bcr-Abl, middle-T antigen, Ksr, PKC, PI-3 kinase, ASK1 kinase, and cdc25C phosphatase. We also identified human Grb10, an adaptor protein with SH2 domain associated with the IGF-IR beta subunit. Smith's laboratory showed that Grb10 preferentially binds to the IR in intact cells. Using the interaction trap screen (active cytoplasmic domain of the IGF-IR) 55PIK and SOCS-2 proteins were also identified. However, 55PIK and SOCS-2 also interact with the IR in the yeast two hybrid system. These studies raise the possibility that 14-3-3 and Grb10 may play a role in insulin and IGF-I signal transduction and may underlie the observed differences. PMID- 10453982 TI - 4-Hydroxy-2,3-nonenal as a signal for cell function and differentiation. AB - 4-Hydroxy-2,3-nonenal (HNE) is a biologically active aldehydic end product of oxidative decomposition of omega-3 and omega-6 polyunsaturated fatty acids of membrane phospholipids, a process referred to as lipid peroxidation. HNE has been detected in several experimental and clinical conditions in which oxidative stress has been reported to occur and several authors have suggested that HNE and related 4-hydroxy-2,3-alkenals (HAKs) of different chain length may act not only as toxic and mutagenic mediators of oxidative stress-related injury but also as biological signals in normal and pathological conditions. In this paper we will review the literature supporting the concept that HNE and HAKs may act as signal molecules able to modulate biological events such as chemotaxis, signal transduction, gene expression, cell proliferation and cell differentiation. PMID- 10453983 TI - Nuclear receptors, their coactivators and modulation of transcription. AB - Nuclear receptors are ligand-dependent transcription factors which can also be activated in the absence of their lipophilic ligands by signaling substances acting on cell membrane receptors. This ligand-independent activation indicates the importance of nuclear receptor phosphorylation for their function. Nuclear receptor-mediated transcription of target genes is further increased by interactions with recruited coactivators forming a novel family of nuclear proteins. CBP/p300, a coactivator of different classes of transcription factors, including the tumor suppressor protein p53, plays a special role acting as a bridging protein between inducible transcription factors and the basal transcription apparatus, and as an integrator of diverse signaling pathways. Coactivators of nuclear receptors and associated proteins forming a multicomponent complex have an intrinsic histone acetylase activity in contrast to nuclear receptor and heterodimer Mad-Max corepressors, which recruit histone deacetylase. Similarly the Rb protein interacts with histone deacetylase to repress transcription of cell cycle regulatory genes. Targeted histone acetylation/deacetylation results in remodeling of chromatin structure and correlates with activation/repression of transcription. Recent data point to the important role of coactivator proteins associated with inducible transcription factors in transcription regulation, and in the integration of multiple signal transduction pathways within the nucleus. PMID- 10453984 TI - Isozymes delta of phosphoinositide-specific phospholipase C. AB - Phospholipase C (PLC, EC 3.1.4.11) is the major starting point in the phosphatidylinositol pathway, which generates intracellular signals that regulate protein kinase C and intracellular calcium concentration. To date, three major types of phosphoinositide-specific PLC species named beta, gamma and delta, have been characterized. This article reviews recent studies on isozymes delta of PLC. Four such isozymes have been cloned and termed delta1-4. Their structural organization, regulation of activity and the interaction with membrane lipid are considered. The intracellular localization of delta isozymes and distribution in various tissues are presented. Attention is given to the pathological conditions in which an abnormal protein level of PLC delta or its activity have been observed. PMID- 10453985 TI - Exogenous sphingosine 1-phosphate and sphingosylphosphorylcholine do not stimulate phospholipase D in C6 glioma cells. AB - In the present study we investigate the effect of exogenous sphingosine, sphingosine 1-phosphate and sphingosylphosphorylcholine on phospholipase D (PLD) activity in glioma C6 cells. The cells were prelabeled with [1-14C]palmitic acid and PLD-mediated synthesis of [14C]phosphatidylethanol was measured. Sphingosine 1-phosphate and sphingosylphosphorylcholine did not stimulate [14C]phosphatidylethanol formation either at low (0.1-10 microM) or high (25-100 microM) concentrations. On the other hand, sphingosine at concentrations of 100 250 microM strongly stimulated PLD activity as compared to the effect of phorbol ester, 12-O-tetradecanoylphorbol 13-acetate (TPA), known as a PLD activator. The effect of TPA on PLD is linked to the activation of protein kinase C. The present study also shows that sphingosine additively enhances TPA-mediated PLD activity. This is in contrast to the postulated role of sphingosine as a protein kinase C inhibitor. These results demonstrate that in glioma C6 cells sphingosine not only affects PLD independently of its effect on protein kinase C, but also is unable to block TPA-mediated PLD activity. PMID- 10453986 TI - The role of cholesterol and sphigomyelin in tyrosine phosphorylation of proteins and capping of Fcgamma receptor II. AB - Cross-linking of cell surface receptors by multivalent ligands, e.g. by antibodies, evokes their clustering -- patching. Subsequently, these clusters can be translocated by the acto-myosin machinery toward one pole of the cell and assembly cap. Patching of FcgammaRII in U937 cells correlates with tyrosine phosphorylation of several proteins while cap assembly correlates with their dephosphorylation. To study the mechanism of activation of tyrosine kinases during FcgammaRII activation we disturbed the organization of the putative plasma membrane microdomains by depletion of membrane cholesterol and sphingomyelin. Cholesterol was removed with the use of beta-cyclodextrin while sphingomyelin was decomposed by exogenous sphingomyelinase. Cyclodextrin at 5-10 mM removed about 70% of cholesterol from the cells and abolished the assembly of FcgammaRII caps thereby arresting the receptors at the patching stage. Similarly, 70 mU/ml sphingomyelinase inhibited cap formation by 60%. Cholesterol and sphingomyelin depletion also suppressed the tyrosine phosphorylation of proteins which accompanied cross-linking of FcgammaRII. The observations indicate that cholesterol and sphingomyelin can control the interactions of tyrosine kinases with clustered FcgammaRII. PMID- 10453987 TI - Insulin-like growth factor I activates insulin receptor substrate 1 and Ras in human osteosarcoma cells. AB - Insulin-like growth factor I (IGF-I) stimulates multiplication of the human osteosarcoma cell line, MG-63, by acting through IGF-I receptor. We have characterized IGF-I stimulated phosphorylation of IRS-1, activation of Ras cycle and phosphorylation of c-Jun in this cell line. Serum starved MG-63 cells were (1) IGF-I stimulated and lysates were immunoprecipitated with polyclonal IRS-1 antibody or (2) metabolically labeled with [32P]orthophosphoric acid and then cells were treated with IGF-I. Cell lysates were immunoprecipitated with p21Ras antibody (Y13-259) and bound nucleotides were analysed by thin-layer chromatography. We demonstrated tyrosine phosphorylation of IRS-1/2 immunoprecipitated from MG-63 cells stimulated with IGF-I. We also showed an increased level of GTP in p21Ras immunoprecipitates from IGF-I treated cells. Nuclear extracts prepared from 32P-labeled cells before and after addition of IGF I were immunoprecipitated with c-Jun antibody. After electrophoresis and autoradiography, phosphorylation of the c-Jun band was seen to be IGF-I independent. Phosphoamino acid analysis of the c-Jun band showed that phosphoserine was the major species. PMID- 10453988 TI - Sphingosine, sphingosylphosphorylcholine and sphingosine 1-phosphate modulate phosphatidylserine homeostasis in glioma C6 cells. AB - The effect of sphingosine, sphingosylphosphorylcholine and sphingosine 1 phosphate on L-[U-14C]serine incorporation into phosphatidylserine and phosphatidylserine-derived phosphatidylethanolamine was investigated in intact glioma C6 cells. Sphingosine, sphingosylphosphorylcholine and sphingosine 1 phosphate are potent signalling molecules which, due to their physicochemical features, may function as amphiphilic compounds. It has been found that sphingosine and sphingosylphosphorylcholine (amphiphilic cations) significantly increase [14C]phosphatidylserine synthesis and decrease the amount of 14C-labeled phosphatidylethanolamine. Sphingosine 1-phosphate (an amphiphilic anion) was without effect on phosphatidylserine synthesis but, similarly as sphingosine and sphingosylphosphorylcholine, reduced the conversion of phosphatidylserine to phosphatidylethanolamine. These results strongly suggest that sphingosine, sphingosylphosphorylcholine and sphingosine 1-phosphate can modulate cellular phospholipid homeostasis by stimulation of phosphatidylserine synthesis and an interference with phosphatidylserine decarboxylase. PMID- 10453989 TI - The decisive role of the water structure in changes of conformation of nucleic acids. AB - This review summarizes data on the structure and properties of water under normal conditions, at high salt concentration and under high pressure. We correlate the observed conformational transitions in nucleic acids with changes in water structure and activity, and suggest a mechanism of conformational transitions of nucleic acid involving these changes. We conclude that the Z-DNA form is induced only at low water activity caused by high salt concentrations and/or high pressure. PMID- 10453990 TI - Nonenzymatic hydrolysis of oligoribonucleotides. V. The elements affecting the process of self-hydrolysis. AB - Chemical instability of some of the phosphodiester bonds, often observed in large RNAs, visualizes the autocatalytic properties of this class of nucleic acids. Unexpectedly, selective hydrolysis occurs also in short oligoribonucleotides (as short as a tetramer or hexamer). Herein, we describe additional experiments which support the conclusion that the hydrolysis is not due to ribonuclease contamination but is of autocatalytic origin and is related to the sequence and structure of single-stranded oligomers. Moreover, we show that the presence in the reaction mixture of polyamines, such as spermidine, is essential for hydrolysis of oligoribonucleotides. PMID- 10453991 TI - A human putative Suv3-like RNA helicase is conserved between Rhodobacter and all eukaryotes. AB - We have cloned and sequenced a cDNA of the human homologue of the Saccharomyces cerevisiae Suv3 putative RNA helicase which is indispensable for mitochondrial function in yeast. The human Suv-3-like protein has a typical mitochondrial leader sequence. Northern blot data and analysis of ESTs in the data banks indicate that this human gene (SUPV3L1) is expressed in practically all tissues, though at different levels. Sequence homology analysis has shown a strong conservation of the protein in a number of eukaryotic organisms -- plants, mammals and fungi, but no close homologues exist in bacteria with the exception of the purple bacterium Rhodobacter sphaeroides. This gene is thus ubiquitously present in all eukaryotic organisms. PMID- 10453992 TI - Thermodynamic contribution of nucleoside modifications to yeast tRNA(Phe) anticodon stem loop analogs. AB - The determination of the structural and functional contributions of natural modified nucleosides to tRNA has been limited by lack of an approach that can systematically incorporate the modified units. We have produced a number of oligonucleotide analogs, of the anticodon of yeast tRNA(Phe) by, combining standard automated synthesis for the major nucleosides with specialty chemistries for the modified nucleosides. In this study, both naturally occurring and unnatural modified nucleotides were placed in native contexts. Each oligonucleotide was purified and the nucleoside composition determined to validate the chemistry. The RNAs were denatured and analyzed to determine the van't Hoff thermodynamic parameters. Here, we report the individual thermodynamic contributions for Cm, Gm, m1G, m5C, psi. In addition m5m6U, m1psi, and m3psi, were introduced to gain additional understanding of the physicochemical contribution of psi and m5C at an atomic level. These oligonucleotides demonstrate that modifications have measurable thermodynamic contributions and that loop modifications have global contributions. PMID- 10453993 TI - Damaged DNA-binding proteins: recognition of N-acetoxy-acetylaminofluorene induced DNA adducts. AB - Proteins which bind to the DNA damaged by genotoxic agents can be detected in all living organisms. Damage-recognition proteins are thought to be generally involved in DNA repair mechanisms. On the other hand, the relevance to DNA repair of some other proteins which show elevated affinity to damaged DNA (e.g. HMG-box containing proteins or histone H1) has not been established. Using the electrophoretic mobility-shift assay we have investigated damage-recognition proteins in nuclei from rat hepatocytes. We detected two different protein complexes which preferentially bound the DNA damaged by N-acetoxy acetylaminofluorene. One of them also recognized the DNA damaged by benzo(a)pyrene diol epoxide (yet with much lower efficiency). The proteins which bind to damaged DNA are permanently present in rat cells and their level does not change after treatment of animals with the carcinogens. Differences in the affinity of the detected damage-recognition proteins to DNA lesion evoked by either carcinogen did not correlate with more efficient removal from hepatic DNA of 2-acetylaminofluorene-induced adducts than benzo(a)pyrene-induced ones. PMID- 10453994 TI - Heterologous expression of genes in filamentous fungi. AB - Isolation of some biologically important proteins from natural sources was found to be too expensive or scarcely possible (human proteins). The problem could be solved by expression of heterologous genes. Many biologically active proteins have been successfully expressed in filamentous fungi, some of them, however, at a low level. Thus, improvement of this technique appears to be a very important task. The process comprises several steps. Some of them, such as efficient transformation, vector construction, processing of signal sequences, post translational modifications and secretion of the expressed proteins, have been intensively investigated. This review presents obstacles and problems encountered in expression of heterologous genes and discusses strategies of development in this area. PMID- 10453995 TI - Expression of ras-family genes in the cell cycle and during differentiation of the lower eukaryote Physarum polycephalum. AB - Messenger RNA levels of three ras-family genes (Ppras1, Ppras2, and Pprap1) were measured in different life forms and throughout the cell cycle of the slime mold Physarum polycephalum. All three genes are expressed at constant rates in the uninucleate amoebae and flagellates, regardless of the culture conditions (solid or liquid medium, particulate or dissolved nutrients). In the multinucleate stages (micro- and macroplasmodia) Ppras1 and Pprap1 mRNAs are somewhat less abundant, while Ppras2 is not expressed at all. The early stages of the amoeba plasmodium transition proceed without any drop in Ppras2 expression. During the synchronous cell cycle in macroplasmodia Ppras1 and Pprap1 are expressed at a constant level. PMID- 10453996 TI - 10-Undecynoic acid, an inhibitor of cytochrome P450 4A1, inhibits ethanolamine specific phospholipid base exchange reaction in rat liver microsomes. AB - 1,12-Dodecanedioic acid, the end-product of omega-hydroxylation of lauric acid, stimulates in a concentration dependent manner, phosphatidylethanolamine synthesis via ethanolamine-specific phospholipid base exchange reaction in rat liver endoplasmic reticulum. On the other hand, administration to rats of 10 undecynoic acid, a specific inhibitor of omega-hydroxylation reaction catalyzed by cytochrome P450 4A1, inhibits the ethanolamine-specific phospholipid base exchange activity by 30%. This is accompanied by a small but significant decrease in phosphatidylethanolamine content in the endoplasmic reticulum and inhibition of cytochrome P450 4A1. On the basis of these results it can be proposed that a functional relationship between cytochrome P450 4A1 and phosphatidylethanolamine synthesis exists in rat liver. Cytochrome P450 4A1 modulates the cellular level of lauric acid, an inhibitor of phospholipid synthesis. In turn, ethanolamine specific phospholipid base exchange reaction provides molecular species of phospholipids, containing mainly long-chain polyunsaturated fatty acid moieties, required for the optimal activity of cytochrome P450 4A1. PMID- 10453997 TI - Phosphorylation of yeast ribosomal proteins by CKI and CKII in the presence of heparin. AB - We have found that heparin has a different effect on Trichosporon cutaneum ribosomal protein phosphorylation by CKI and by CKII. In the presence of heparin, modification of 13 kDa, 19 kDa and 38 kDa proteins catalyzed by CKII was inhibited, while in the case of CKI, in addition to protein of 15 kDa, phosphorylation of 20 kDa and 35 kDa proteins was detected. It was also found that, in the presence of heparin, phosphorylation of P proteins (13 kDa and 38 kDa) by ribosome-bound protein kinases was inhibited. Moreover at the same conditions modification of 40 kDa protein was observed in all four yeast species tested. PMID- 10453998 TI - Modulatory effect of divalent metal cations on the phosphotyrosine activity of the frog liver acid phosphatase. AB - Frog liver acid phosphatase hydrolyzes phosphotyrosine at acidic pH optimum. Mn2+, Ca2+ and Mg2+ (but not Zn2+) ions modulate this activity by shifting its pH optimum to physiological pH. This effect is not observed when p nitrophenylphosphate is used as a substrate. Phosphoserine and phosphothreonine are not hydrolyzed under the same conditions. PMID- 10453999 TI - Plasma interference in an enzyme-linked immunosorbant assay using a commercial matched antibody pair. AB - In this study, severe plasma interference was repeatedly documented in an IL-1ra sandwich enzyme-linked immunosorbant assay (ELISA) using a commercial matched antibody pair. Several physical and biochemical treatments were used in an attempt to alleviate this plasma effect including the following: buffer optimization, sample dilution, increasing incubation temperature, heat treatment of plasma, increasing detergent concentrations, glutaraldehyde pretreatment of the plate and the addition of polyethylene glycol (PEG). Evaluation of several buffers demonstrated that the range of optical densities could be increased dramatically with the use of an appropriate buffer. Of the treatments examined, only the addition of polyethylene glycol (PEG) to the dilution buffer created a marked improvement in the ELISA, despite a resulting background increase. Further investigation demonstrated that 10% PEG in the dilution buffer added to biotinylated antibody and the streptavidin provided the greatest improvement to the sensitivity of the ELISA. PMID- 10454000 TI - Platelet-activating factor plays a role in the mechanism of major histocompatibility complex in T lymphocytes. AB - In recent studies, using a swine model of single lung transplantation, we demonstrated that IRI alone increased MHC II expression in the host's peripheral T lymphocytes. The inhibition of increased MHC II expression with TCV-309, a specific platelet-activating factor (PAF) antagonist suggested that PAF might play a role in the mechanism of increased MHC II expression. The purpose of the current study was two fold: 1) to investigate the mechanism of PAF-induced increased expression of MHC II in T lymphocytes, 2) to determine whether a specific PAF-antagonist, TCV-309, is capable of inhibiting the increased expression in an in vitro system. This study was subdivided, using four in vitro conditions: 1) purified resting T cells, 2) purified proliferating T cells, 3) PBL treated with PAF, and 4) PBL preincubated with TCV-309 and treated with PAF. The level of MHC II on T cells were measured by two color flow cytometry analysis (swine anti-CD3, MHC II-DR-(beta)antibodies). Both MHC II intensity and the number of CD3+MHC+ T cells did not change in resting purified T cells once treated with PAF, Furthermore, MHC II intensity did not change in purified proliferating T cells treated with PAF. The number of CD3+MHC+ T cells, however, increased significantly (p<0.05) from day 1 to day 4 as compared with pre treatment value (day 0) for purified proliferating T cells. Treatment of PBL with PAF (10(-7)M) resulted in a significant (p<0.05) increase in MHC II expression from day 2 to day 4 post-treatment. The number of CD3+MHC+ T cells in PBL, however, did not change significantly upon treatment with PAF. The results of this study indicated that PAF did not have a direct effect on increased MHC II expression in resting or proliferating purified T lymphocytes. However, the mechanism of PAF-induced increased expression of MHC II in T cells may be via an indirect pathway involving accessory cells. TCV-309, a specific PAF receptor antagonist, is capable of inhibiting this PAF-induced increased expression of MHC II in T cells. PMID- 10454001 TI - Optimisation of a technique for isolating lymphocyte subsets from human endometrium. AB - Human endometrium is a rich source of lymphocytes which may have unique immunoregulatory functions. The aim of this study was to compare current procedures for endometrial tissue disaggregation, and optimise a method for isolation of endometrial lymphocytes. Tissue was obtained from 41 women undergoing elective hysterectomy or dilation and curettage (D&C) for reasons of benign non-endometrial pathology. Specimens were exposed to reduction/chelation, mechanical or enzymatic disruption. Optimal single cell suspensions of high yields (mean 8.8 x 10(6) range 3.5-18 x 10(6)lymphs) and good viability (60%) were obtained, using a combination of collagenase IV (200 U/ml) and DNase I (35 U/ml). Suspensions were further purified by density gradient centrifugation. Multi-colour flow cytometry was used for analysis of endometrial lymphocyte subsets. Cell suspensions were stained with mAbs specific for CD3, CD4, CD8, CD56, CD45 and CD14, and it was clearly shown that the developed method had no effect on surface glycoprotein expression. Phenotypic analysis revealed consistent populations of endometrial large granular lymphocytes (CD56+CD3-) 54.16%, and T-cells (CD3+) 37.73%. This technique was applicable to the characterisation of T-cell populations, including CD8+ (56.6%), CD4+ (44.0%), and particularly smaller populations of CD4+CD8+(3.56%), CD4-CD8-(3.34%) and CD56+(6.3%) due to it's sensitivity. In conclusion, optimised enzymatic digestion, in combination with flow cytometry provides an effective method for phenotypic examination of small endometrial lymphocyte subpopulations. PMID- 10454002 TI - Sex-related immune changes in young mice. AB - Here we describe changes in selected immune parameters related to age and sex in young mice. We focused on the T cell compartment and studied thymuses and spleens from mice 3 to 9 weeks of age in order to bracket the time period around murine puberty. With regard to distribution of immune cells, no significant sex-related changes were seen in thymocyte expression of CD3, CD4, CD8, or CD4/CD8 or splenocyte expression of CD3, CD4, CD8, or CD45R/B220, a pan B cell marker. For splenocytes, significantly more cells were positive for CD3 in older (6-9 week old) compared with younger (3-4 week old) mice. Splenocyte and thymocyte cell proliferation as measured by DNA synthesis in response to in vitro mitogens was compared for cells from male and female mice over the ages studied. Thymocyte proliferation was not related to age or sex of the mice. For splenocytes of the youngest mice (3 weeks old), the response to a cell surface-receptor-independent mitogenic combination of phorbol ester and ionomycin induced a significantly greater response in cells from female mice compared with male mice. This trend was reversed for mice of 4-6 weeks of age, where the response by splenocytes from males was significantly greater than that by cells from females. For mice 7-8 weeks of age, splenocytes from female mice responded significantly less to stimulation by antibody to CD3, a component of the T-cell receptor. Our results demonstrate that depending on the assays employed, sexual dimorphism in the immune system may be demonstrated prior to puberty. PMID- 10454003 TI - Identification of immunodominant antigens in Mexican strains of Trypanosoma cruzi. AB - Chagas disease represents an important public health problem. In Mexico most studies have been performed using Trypanosoma cruzi' antigens extracted from strains of other geographical origins. This work was aimed at developing a reactive antigen to perform serological diagnosis of Chagas' disease, using Mexican T. cruzi strains. We prepared antigenic extracts from epimastigotes, trypomastigotes and sphaeromastigotes of three Mexican strains. Parasites homogenate was obtained by lysis and sonication, solubilized proteins were analyzed by SDS-PAGE, Western-blot assays, and ELISA to determine the reactivity against sera from chagasic reference serum and chagasic and leishmaniasic patients and healthy donors. Western Blot profiles revealed, with the reference serum, eleven main components between 212 to 25 kDa; however, five bands corresponding to 74, 44, 31, 25 and 18 kDa antigens were recognized by the T. cruzi reactive sera from Mexican chagasic patients, which did not cross-react with Leishmania mexicana. Antigens from the Tequesquitengo strain yielded the best reactivity in the enzymatic immunoassay, thus enabling us to propose their use for serodiagnoses of Chagas' disease in Mexico. PMID- 10454004 TI - Rapid onset monoclonal gammopathy in cutaneous lupus erythematosus: interference with complement C3 and C4 measurement. AB - Accurate nephelometric immunoassay requires both the analyte and the calibration standard to have the same molecular mass. Alteration in the size of the analyte can affect light scatter and yield erroneous results. We report a case where an autoantibody, a monoclonal IgM with immunoconglutinin activity, interfered with the nephelometric quantitation of plasma complement components C3 and C4. PMID- 10454005 TI - Molecular investigations implicate human endogenous retroviruses as mediators of anti-retroviral antibodies in autoimmune rheumatic disease. AB - Polymerase chain reaction using specific primers, failed to detect HTLV-I amplicons in patients with rheumatic diseases previously shown to possess antibodies to retroviral products. However, by employing broad spectrum oligonucleotide primers, 135 bp amplicons were generated from peripheral blood mononuclear cells and synovial fluid cells. Subsequent cloning and DNA sequencing revealed homology to a number of exogenous and human endogenous retroviruses (HERVs). Furthermore, in combining the presence of type B and C related endogenous retroviruses, a significant association (p=0.014) was apparent for chronic autoimmune rheumatic diseases as compared to controls. Reverse transcription polymerase chain reaction of RNA derived from patients, healthy controls and cell lines (U937, BJAB, human endothelial lung fibroblasts) demonstrated ubiquitous expression of HERV-K10 and RTVL-H2. Furthermore messenger RNA expression of HERV-K10 was enhanced in fibroblasts infected with human cytomegalovirus. It is plausible that subsequent production of HERV peptides could explain the presence of anti-retroviral antibodies in cohorts of patients with autoimmune rheumatic diseases. PMID- 10454006 TI - Adsorption of glycosaminoglycans onto coral--a new possible implant biomaterials for regeneration therapy. AB - The adsorption of glycosaminoglycans (heparin, heparan sulfate, dermatan sulfate, highly sulfated chondroitin sulfate, chondroitin sulfate, and hyaluronan) onto coral has been investigated. Granules of natural coral of specific diameter, between 100 and 500 microm, having high content of calcium (> 98%) and a homogeneous surface adsorb glycosaminoglycans with different capacity. Heparin (maximum adsorption 1.29 +/- 0.10 mg/20 mg of coral, 6.45% w/w) is adsorbed more than highly sulfated chondroitin sulfate species (maximum adsorption of 0.90 +/- 0.06 mg/20 mg of coral, 4.50% w/w), chondroitin sulfate (maximum adsorption of 0.72 +/- 0.06 mg/20 mg of coral, 3.60% w/w), dermatan sulfate (maximum adsorption of 0.70 +/- 0.06 mg/20 mg of coral, 3.50% w/w) and heparan sulfate (maximum adsorption of 0.72 +/- 0.07 mg/20 mg of coral, 3.60% w/w). Hyaluronan is not adsorbed onto granules of coral. The percentage adsorption of polyanions onto coral depends mainly on their charge density, with sulfate groups being more important than carboxyl groups. This study found no evidence that iduronic acid is more important than glucuronic acid and no role of molecular mass on the adsorption of polysaccharides onto coral was found. The adsorption of glycosaminoglycans is driven by electrostatic interactions with calcium sites of coral that are dependent on pH and blocked in the presence of large amounts of salt. Due to these peculiar properties, the combination of granules of natural coral with glycosaminoglycans makes this material potentially useful in osseointegration in bone metabolism or periodontal therapy. PMID- 10454007 TI - Anti-inflammatory properties of triblock siloxane copolymer-blended materials. AB - Implantable biomaterials often trigger a variety of adverse responses. Because polydimethyl siloxane surfaces have good hemo- and bio-compatibility, it is generally believed that surface biocompatibility may be improved by modifying biomaterial surfaces with silicone-like properties. For this, we developed a series of polycaprolactone-polydimethylsiloxane-polycaprolactone (PDMS-PCL) copolymers. By mixing the substrate material--polyvinyl chloride--with low concentrations (1.2 and 2.4%) of the PDMS-PCL copolymer, we generated materials with silicone-like surface properties as reflected by increased surface silicon content and surface contact angles. We assessed the biocompatibility of these surfaces in vitro and found that the addition of PDMS-PCL significantly reduced the percentages of surface-'denatured' fibrinogen, a critical element of genesis of many adverse responses to implanted biomaterials. Indeed, using an animal implantation model, we find that PDMS-PCL-blended materials triggered significantly weaker inflammatory responses than did polyvinyl chloride, the substrate control. The results from these experiments suggest that the use of PDMS-PCL additives (2.4%) in polymer blends is a useful means of camouflaging the substrate surface properties and improving the biocompatibility of biomaterials. PMID- 10454008 TI - Highly crosslinked, PEG-containing copolymers for sustained solute delivery. AB - Novel ionizable polymer networks were prepared from oligo(ethylene glycol) (OEG) multiacrylates and acrylic acid (AA) employing bulk radical photopolymerization techniques. The properties of these materials exhibited a complex dependence on the network structure and composition, and the materials were therefore used in the design of controlled release devices with precisely controlled properties. The release kinetics of model solute proxyphylline exhibited a strong compositional dependence, with measured diffusion coefficients varying over several orders of magnitude, depending on the polymer network structure and the pH of the release medium. Varying the OEG chain length provided a means of coarsely adjusting the proxyphylline release rate, while varying the AA content and the pH offered a more precise measure of control. PMID- 10454009 TI - A DSC study of the miscibility of poly(ethylene oxide)-block-poly(DL-lactide) copolymers with poly(DL-lactide). AB - The purpose of this study was to examine the miscibility of poly(ethylene oxide) block-poly(DL-lactide) copolymers with poly (DL-lactide). The copolymers L7E73L7 and L17E78L17 (L = carbonyloxymethylmethylene unit, OCOCH(CH3); E = oxyethylene unit, OCH2CH2) were synthesised by non-catalysed anionic polymerisation and characterised by gel permeation chromatography and 13C NMR. Blends of each of the copolymers with poly(DL-lactide) with compositions over the range from 10 to 90 wt% copolymer were cast as thin films and examined by differential scanning calorimetry (DSC) to determine glass transition temperatures (Tg) and melting temperatures (Tm). The phase diagram showed a region of miscibility above the melting point of the copolymer in the system (approx. 35-40 degrees C). Within this region the system was glassy at low mass fractions of oxyethylene in the copolymer (wE < or = 0.1) and rubbery at higher mass fractions. Below Tm a mechanically compatible glassy blend existed at low wE whilst quenching of systems of higher wE led to phase separation, the biphasic region consisting of crystalline Em-sequences of copolymer separated from non-crystalline poly(DL lactide). The phase diagram resulting from this study provides the means for the design of drug delivery systems based on blends of poly(DL-lactide) and poly(ethylene oxide)-containing components. The crystal melt boundary can be lowered by the use of block copolymers with short poly(ethylene oxide) blocks permitting the preparation of blends which are miscible at room temperature and rubbery or glassy according to composition. PMID- 10454010 TI - Transformation of modified brushite to hydroxyapatite in aqueous solution: effects of potassium substitution. AB - Brushite (dicalcium phosphate dihydrate, DCPD, CaHPO4 x 2H2O) was deposited electrolytically from calcium dihydrogen phosphate solution with and without potassium chloride (KCl) as a supporting electrolyte. The kinetics of brushite transformation to hydroxyapatite (HA, Ca5(PO4)3OH) in the presence of calcium and protein free, aqueous body fluid (Hank's balanced salt solution, HBSS) was investigated. We present evidence that the deposition of brushite in a KCl supported electrochemical bath yields a modified brushite where some of the calcium is substituted by potassium. Transformation of both normal (i.e. potassium free) and modified brushite to hydroxyapatite upon exposure to calcium and protein-free aqueous fluid (HBSS) was followed by reflectance Fourier Transform Infrared (FTIR) spectroscopy and X-ray diffraction (XRD) techniques. Changes in the morphology of the coatings were studied using scanning electron microscopy (SEM). Results indicate that modified brushite undergoes faster transformation to hydroxyapatite in HBSS in comparison to normal brushite. Our results show that the presence of potassium ions in the brushite not only favors the formation of different intermediate phases but also alters transformation rates to HA. PMID- 10454011 TI - Application of dissolution experiments to characterise the structure of pulsed laser-deposited calcium phosphate coatings. AB - A dissolution test was performed with pulsed laser (Nd: YAG, 355 nm)-deposited calcium phosphate coatings composed of hydroxyapatite (HA) and alpha-tricalcium phosphate (alpha-TCP) in different proportions, as a result of the use of different deposition rates. During immersion in a Ca2+-free Hank's solution, the dissolution kinetics were determined while other structural and compositional properties of the coatings were derived. It was possible to infer that the alpha TCP is distributed uniformly and that the coating is of a non-columnar compact grain structure. The mass ratio of the phases for each coating was also determined and was related to the X-ray diffraction intensities. When incomplete, the hydroxylation level of the HA in the coatings is completed after immersion. PMID- 10454012 TI - Accelerating effects of chitosan for healing at early phase of experimental open wound in dogs. AB - Chitosan is a polymeric beta(1 --> 4) glucosamine (2-amino-2-deoxy-D-glucose) and N-acetyl-D-glucosamine (2-acetamido-2-deoxy-D-glucose) which has been reported as a wound healing accelerator. In order to evaluate the efficacy of chitosan as an accelerator of wound healing, experimental open skin wounds were made on the dorsal side in three normal beagles. Cottonfiber-type chitosan (degree of acetylation = 18%) was applied for 15 days, and the process of wound healing was evaluated histologically and immunohistochemically. On day 3 postwounding, the chitosan-treated wounds showed histologically severe infiltration of polymorphonuclear (PMN) cells and an increase in effusion compared with that in the control. Granulation was more pronounced by the chitosan treatment on day 9 and 15 postwounding. Immunohistochemical typing of collagen I, III and IV showed increase of the production of type III collagen in the chitosan group. The appearance of mitotic cells occurred numerously in the control on postwounding day 3, and in the chitosan group on postwounding day 6. These results suggest chitosan to be having a function in the acceleration of infiltration of PMN cells at the early stage of wound healing, followed by the production of collagen by fibroblasts. PMID- 10454013 TI - Bioactive ceramics prepared by sintering and crystallization of calcium phosphate invert glasses. AB - Novel glass-ceramics were synthesized via sintering and crystallization by heating powder compacts of SiO2-free calcium phosphate invert glasses of 60CaO x 30P2O5 x 7Na2O x 3TiO2 or 60CaO x 30P2O5 x 7Na2O x 3MgO at 800-850 degrees C in air. The glass-ceramics were relatively dense materials consisting of crystalline phases such as beta-Ca3(PO4)2 and beta-Ca2P2O7 with glassy phases. The compacts were densified by the viscous flow of the glassy phases while heating. By soaking in simulated body fluid at 37 degrees C, a calcium phosphate phase was formed newly on the surface of the glass-ceramic derived from 60CaO x 30P2O5 x 7Na2O x 3TiO2 glass, while the phase was not formed on that derived from 60CaO x 30P2O5 x 7Na2O x 3MgO glass: the former was implied to show bioactivity. Composition of the glassy phase as the matrix varies with the additives such as TiO2 and MgO, and the chemical properties of the phase influence the bioactivity of the glass ceramics. The glass-ceramic derived from 60CaO x 30P2O5 x 7Na2O x 3TiO2 glass has relatively high fracture toughness of K(IC) approximately 2 MPa m(0.5) and bending strength of 100-120 MPa. PMID- 10454014 TI - Functionally graded fluoridated apatites. AB - Fluoridated hydroxyapatite was synthesized at 80 +/- 1 degree C and pH 7.4 +/- 0.2 using a gradient fluoride supply system. X-ray diffraction analysis showed a typically apatitic pattern, although the (3 0 0) reflection was broader than that of homogeneous fluorapatite. Scanning electron micrographic observation indicated that the apatite was composed of rod-like crystals similarly to fluorapatite. High-resolution transmission electron microscopy showed electron damage in the core of the crystal. When the apatite pellet was prepared, electron spectroscopy for chemical analysis showed a negative gradient of fluoride concentration with depth in the crystals. The apparent solubility in 0.5 mol/l acetate buffer solution (37 degrees C and pH 4.0) was 9.16 +/- 0.39 mmol/l, much less than that of homogeneous hydroxyapatite 32.3 +/- 1.9 mmol/l, and less than that of heterogeneous two-layer fluoridated apatite with an outer fluoride-rich layer 12.5 +/- 0.6 mmol/l, which was synthesized previously by supplying fluoride during the latter half of the experimental period. These results suggest that graded fluoridated apatite may be formed by this process and have higher acid resistance than two-layer fluoridated apatite. PMID- 10454015 TI - Physico-chemical characterization of Ca-alginate microparticles produced with different methods. AB - In the present paper the physico-chemical characterization of Ca-alginate microparticles produced with different methods is presented. Ca-alginate microparticles were obtained either by emulsification method or by dripping an aqueous alginate solution into a solution of calcium salt. Inverse Size Exclusion Chromatography (ISEC) was used for the determination of dimensions of the pores and porous volume of microparticles having a mean diameter of 220 microm when obtained by emulsification method. The same technique was used to study the variation of the pore size and porous volume with pH. The results were related with the content of calcium and sodium in the microparticles, before and after their treatment with different HCl solutions. For the particles with a mean diameter of 1.2 mm (obtained by dripping method) we adopted an other approach based on the steric exclusion of solute at equilibrium. For a convenient interpretation of the obtained data, determination of water regain, swelling degree, and scanning electron microscopy (SEM) were performed. Finally, a comparison of the characteristics of microparticles produced by ionic and epichlorohydrin crosslinkings was made. The maximum dimensions of the pores of the microparticles obtained by emulsification were found smaller than those obtained by other technics. The variation of the dimensions of the pores and porous volume with pH is not significant. The structure of the chemically crosslinked beads with epichlorohydrin is more elastic and the swelling is reversible; after drying and reswelling process, the dimensions of the pores and porous volume of these microparticles remain unchanged. On the opposite, for the microparticles obtained by emulsification or dripping method in the presence of calcium ions, these characteristics are changed after a first drying process. PMID- 10454017 TI - Quercetin inhibits LPS-induced nitric oxide and tumor necrosis factor-alpha production in murine macrophages. AB - High amounts of nitric oxide and TNF-alpha generated by activated macrophages induce several pathophysiological conditions during acute and chronic inflammation. Identification of new pharmacological reagents that can prevent TNF alpha and/or NO overproduction is of considerable medical interest. In this report we provide evidence that the overproduction of TNF-alpha and NO by LPS stimulated macrophages can be markedly inhibited by quercetin, a major active component of plant Rhododendron cinnabarium. PMID- 10454016 TI - Urinary neopterin and kynurenine in patients submitted to surgical stress with different inhalational anesthetics (halothane or isoflurane). AB - Measurements were taken of urinary levels of neopterin (NPT) and kynurenine (KYN), using an HPLC method for their simultaneous analysis in patients submitted to anesthetical surgical stress with two different inhalational anesthetics (halothane and isoflurane). We studied twenty-one women affected by uterine fibromyomatosis and submitted to total hysterectomy (mean age of 42.7+/-5.4 years). They showed the same pre-operative evaluation (ASA-1), and underwent the same i.v. anesthetic treatment. Our patients were randomized in two groups: Group A: 11 patients had halothane as an inhalational anesthetic drug for the maintenance of the anesthetic induction (mean time= 1 h). Group B: 10 patients had isoflurane. A significant decrease in urinary NPT and KYN, parallel to serum NPT, was found 4 h after anesthetic induction. Raised NPT levels appeared 24 h after A.I. with significant increased levels after 7 days. A strong correlation between urinary and serum NPT levels was seen (Rs= 0.74; p < 0.001). Significantly low KYN levels were observed both 4 h and 24 h after A.I.. In addition to the delayed increase of the excretory KYN levels, significantly raised KYN levels in Group B (isoflurane) 48 h after A.I. (10.59+/-14.31 vs 5.99+/-7.17 micromol/mol creat.; p < 0.01) were shown, whereas in Group A (halothane) we observed a progressive increase as compared to the pre-surgery values starting from 72 h after surgery. Our data seem to show that: (a) it is possible to have a biochemical and non invasive monitoring of the anesthetical surgical stress on MM "priming" activity; (b) the activation of the phagocyte compartment is one of the earlier immunological events after surgery (NPT), but the efficiency of this "priming" appears to be delayed (KYN); (c) isoflurane appears to induce an earlier recovery in MM activation. PMID- 10454018 TI - Effects of fentanyl on cellular immune functions in man. AB - In order to analyze the effects of the opioid agonist fentanyl on cells of the innate immune system, seven healthy individuals were treated intravenously with the opioid fentanyl and five subjects received a placebo. Respiratory burst of polymorphonuclear cells (PMNC) and phenotypes of peripheral blood lymphocytes (PBL) were analyzed from blood samples drawn before, 15 and 30 min after fentanyl or placebo application. In addition, in vitro effects of fentanyl on natural killer (NK) activity was assessed. Fentanyl administration affected neither superoxide production of PMNC nor circulating numbers of B-and T-lymphocytes. In contrast, NK cell (CD16+/CD56+) numbers significantly increased in response to fentanyl. However, no direct influence of fentanyl on NK cell function in vitro could be detected. These results suggest a transient effect of fentanyl on NK cell circulation which seemed to be centrally mediated rather than a direct effect of this opioid on NK cells. PMID- 10454019 TI - Early metabolic changes in peripheral blood cells of renal transplant recipients treated with cyclosporine A. AB - We histochemically examined (phosphatase acid-AcP, phosphatase alkaline-ALP, succinate dehydrogenase-SDH, lactate dehydrogenase-LDH) the peripheral blood of renal transplant recipients and controls before (day 0) and after Cyclosporine A (CsA) treatment (days 1, 2, 7 and 30). We wanted to detect the metabolic changes induced in the CsA resistant cells (leucocytes) by CsA and to evaluate the early effects determined by the drug. There was no difference in enzyme activities between the control group and renal patients before CsA treatment (day 0). AcP and ALP activity increased 1 day after CsA administration and became similar to the control by the day 30. LDH activity increased one day after CsA treatment and remained high during the treatment period (30 days), while SDH activity did not change. These enzymatic variations may suggest that the LDH enzyme is involved in the drug degradation as are other phosphatase and oxidoreductase enzymes (i.e. cytochrome P450). Moreover, the high activity of LDH, the enzyme responsible for interconversion of pyruvate in lactic acid, would indicate that anaerobic glycolysis is preferentially used in the pyruvate pathway. However, SDH did not seem to be directly involved in CsA metabolism. Our findings showed that the CsA treatment induced clear variations of the activity of the cellular phosphatase and oxidoreductase enzymes from the first days of drug administration. The variation of the enzymes studied and the appearance time and duration of the metabolic changes, may be markers of the cellular stress due to CsA internalization. PMID- 10454020 TI - Cocaine effects on immunocompetent cells: an observation of in vitro cocaine exposure. AB - This study investigated in vitro effects of cocaine on the function of T and B lymphocytes, natural killer cells and macrophages in a mouse model. In mature C57BL/6J mice (60-90 day-old), splenocytes were cultured with cocaine at different concentrations ranging from 4 to 64 microg/ml for 24 h. The exposure to cocaine in vitro was found to affect (1) T cell function, with reduced responses to stimulation of Con-A, PHA and Interleukin 2, and decreased production of gamma IFN; (2) B cell function, with reduced response to LPS; (3) natural killer cell function, with attenuated killing capacity; (4) monocyte-macrophage function, with decreased ability to inhibit the growth of tumor cells. The results of the study indicated a direct cocaine effect on four major immune competent cells, and the generally suppressive effects of in vitro cocaine exposure may be related to its in vivo action on the immune system. PMID- 10454021 TI - Facing the future. PMID- 10454022 TI - Squamous carcinoma of the tongue: review. AB - Carcinoma of the tongue is a common condition treated by oral and maxillofacial surgeons. The tongue is a complex anatomical site at the entrance of the oropharynx, bounded by the floor of mouth and lingual mandibular cortex; its form and mobility are crucial for efficient swallowing; speech, and the appreciation of taste. Single and multiple treatments have been used, but the tongue remains a difficult area to assess and treat. In this paper, we have attempted to review some of the papers published over the last 20 years with particular reference to prognosis and functional outcome. We have dealt exclusively with squamous cell carcinoma of the tongue and its management by surgery, radiotherapy, brachytherapy, photodynamic therapy, and chemotherapy. PMID- 10454023 TI - Histological distribution of cervical lymph node metastases from intraoral/oropharyngeal squamous cell carcinomas. AB - The histological frequency and distribution of cervical lymph node metastases was studied in 326 neck dissections from 253 patients with an intraoral/oropharyngeal squamous cell carcinoma. Metastasis was evident in 118 patients (47%) and 18 had bilateral metastasis. For primary sites other than the tongue, metastasis developed initially in a node(s) in the first drainage group (level I or II) with progressive involvement of neighbouring nodes ('overflow'). Simultaneous bilateral metastases were seen in some tumours of the floor of mouth, tongue and oropharynx which involved the midline. An erratic distribution of metastases suggestive of 'fast-tracking' (skip lesions and peppering) was only seen in tongue tumours. The pattern of metastatic spread indicates that level IV nodes must be included in staging and therapeutic neck dissections in tongue cancer. Metastasis to 'accessory' lymph nodes (lingual, buccal) occurs infrequently but may explain some 'local' recurrences. PMID- 10454024 TI - Micrometastasis in oral/oropharyngeal squamous cell carcinoma: incidence, histopathological features and clinical implications. AB - The prognostic importance of accurate pathological staging of oral cancer patients with established lymph node metastases is well known. The present study sought to determine the incidence and clinical significance of minimal neck disease (only micrometastases) and to evaluate clinical features of the primary tumour in relation to the pathological metastatic status. Surgical resection specimens from 178 patients with oral/oropharyngeal squamous cell carcinoma were assessed by routine techniques. Micrometastases were defined as intranodal deposits measuring in total no more than 3 mm at any level of sectioning. Fourteen patients (8%) had only micrometastases and 72 (40%) had established nodal metastases. Patients with only micrometastases were younger than those with established metastases, but there were no differences in T site, T stage or sex. The outcome (2-6 years postsurgery) of patients with only micrometastases was similar to those without cervical node metastases. Hence, the main short-term significance of micrometastases seems to be in relation to clinical staging and elective management of the NO neck. PMID- 10454025 TI - T2 carcinoma of the tongue: the histopathologist's perspective. AB - The histological characteristics and extent of tumour spread at the primary site and in the neck are described in a series of 50 patients with a T2 squamous cell carcinoma of the oral tongue. The likely site of origin was lateral tongue in 29 cases and ventral tongue in 21 cases. Simultaneous tumours were evident in four resections and multifocal dysplasia in a further 13. Lymph node metastasis was diagnosed histologically in 27 cases, including one case with bilateral metastases. Comparison of the groups, with and without metastasis, showed significant differences in tumour thickness, tumour diameter, multifactorial histological malignancy grade, vessel invasion and nerve invasion. The number of positive lymph nodes per patient ranged from 1 to 16 (mean 3), and 17 dissections showed spread beyond the nodal capsule. Postoperative follow-up ranged from 1 to 6 years and, at the time of analysis, 18 patients had died of or with disease. Loco-regional failure was related to 'high-risk' features at the primary site and extracapsular spread in the neck. PMID- 10454026 TI - T2 tongue: reconstruction of the surgical defect. PMID- 10454027 TI - Carcinoma of the tongue: the speech therapist's perspective. PMID- 10454028 TI - Pathology of the N0 neck. AB - The incidence, extent and distribution of nodal metastasis is described in 152 neck dissections from patients with an NO neck undergoing surgery for an intraoral/oropharyngeal squamous cell carcinoma. The preoperative N stage had been determined by palpation under general anaesthesia and magnetic resonance imaging. Metastasis was detected histologically in 32 (21%) of the 152 NO necks. The number of positive nodes per NO neck ranged from 1 to 6. In total, 52 positive nodes were found and 29 (56%) measured 10 mm or less in maximum dimension. Twenty-one dissections (66%) contained a single positive node and 24 (75%) showed a single positive anatomical level. Three cases showed 'skipping' of levels within the neck and one case showed 'peppering'. Seventeen (53%) of the 32 positive NO necks and 31 (60%) of the 52 positive nodes contained only 'micrometastases' (deposits < 3 mm). Microscopic extracapsular spread was evident in five NO necks including one case with extracapsular spread at multiple levels. The study concludes that preoperative staging by palpation and routine magnetic resonance imaging cannot be relied upon to detect early cervical metastatic disease, and the topographic distribution of positive nodes indicates that modified neck dissections should include level IV when the primary tumour involves the tongue. PMID- 10454029 TI - Prophylactic plating of the donor site of osteocutaneous radial forearm flaps. AB - We carried out a retrospective study of fracture morbidity of radial forearm osteocutaneous donor sites. During a period of three-and-a-half years, 12 patients had had composite flaps taken, and 5 of them had subsequently fractured the radius. We then elected to plate the radius prophylactically and found that over a period of just over two years, none of the 8 patients who had had prophylactic plating at the original harvesting operation had subsequently fractured the radius. PMID- 10454030 TI - Management of the radial composite donor site: an orthopaedic opinion. AB - Maxillofacial and plastic surgeons have reported on the management of radial composite flap donor sites, but little reference is made to orthopaedic advice. Orthopaedic surgeons manage forearm injuries more often than other specialties and we thought that an orthopaedic consensus was long overdue. The composite radial donor site subsequently fractures in up to 43% of cases. There is no agreement on the optimal management of these difficult injuries and the patient is often referred for orthopaedic advice. Options include plaster-cast immobilization, internal fixation with either a plate or intramedullary nail, and external fixation. Bone grafting may also be required. A postal questionnaire, using two case histories including radiographs, was sent to 100 consultant orthopaedic surgeons in the UK asking how they would manage the donor site primarily and how they would manage a fracture at this site. Fifty-nine adequate replies were received. Generally, six weeks of immobilization in a plaster of Paris (POP) cast was considered sufficient for the initial management of the donor site. In the event of a fracture, internal fixation with a dynamic compression plate with or without a cancellous bone graft was the most common choice. PMID- 10454031 TI - Reminder--necrotizing fasciitis complicated by Waterhouse-Friderichsen syndrome. PMID- 10454032 TI - Citation for the presentation of the Down Surgical Prize 1997 to John Langdon. PMID- 10454033 TI - Citation for the presentation of the De Puy Prize 1997 to Mr Iain Hutchison. PMID- 10454034 TI - Re: Yu et al. Smoking and the development of Warthin's tumour of the parotid gland. PMID- 10454035 TI - Cervical spine injury in maxillofacial trauma. PMID- 10454036 TI - Re: Planning the consultant workforce in oral & maxillofacial surgery. PMID- 10454037 TI - Re: Editorial 'Should there be a new specialty of head and neck surgery?'. PMID- 10454038 TI - Re: Loescher & Robinson. The effect of surgical medicaments on peripheral nerve function. PMID- 10454039 TI - Gelatin behaviour in dilute aqueous solution: designing a nanoparticulate formulation. AB - Although it has been claimed that nanoparticles can be produced from gelatin, a naturally occurring polypeptide, the commercial conversion of animal collagen to gelatin results in a heterogeneous product with a wide molecular-weight range. This is probably responsible for the widely observed variation in the experimental conditions required for nanoparticle formation. In this study, 0.2% w/v aqueous B225 gelatin solutions were incubated under various conditions of time, temperature, pH and ethanol concentration and characterized by both size exclusion high-performance liquid chromatography (HPLC) and dynamic light scattering. Gelatin was shown to be denatured when the temperature was increased to 37 degrees C (approx.) and the rate of renaturation was optimized over the temperature range 7-20 degrees C at pH 5.0, equivalent to the isoelectric point (IEP). The molecular-weight profile remained unchanged at 37 degrees C (approx.) in the pH range 5-7. When the gelatin solutions were mixed with ethanol, higher molecular-weight fractions (microgel, delta and zeta fractions, all with molecular weights > 700 kDa) precipitated at ethanol concentrations lower than those required to precipitate the lower molecular weight material ( < 700 kDa), with maximum precipitation occurring close to the isoelectric point (pH 5.0). The molecular weight profile of gelatin in solution is evidently critically affected in a time-dependent manner by both pH and temperature. These two factors influence the noncovalent interactions responsible for the molecular structure of gelatin. The molecular weight profiles, in turn, affect the phase behaviour of gelatin in hydroalcoholic solutions. Systematically investigating the effect of time, temperature, pH and ethanol concentration on the molecular-weight distribution profile of a gelatin solution enabled a robust method to be developed for the preparation of colloidal dispersions of non-aggregated gelatin nanoparticles 220-250 nm in diameter. This contrasts with the multiparticulate aggregates produced by earlier literature methods. PMID- 10454040 TI - Osmotic behaviour of polyhedral non-ionic surfactant vesicles (niosomes). AB - In addition to common spherical non-ionic surfactant vesicles (niosomes), disc like, tubular, and polyhedral niosomes have also been reported. The permeability and osmotic activity of niosomes are important in determining their use as controlled-release drug-delivery systems. These properties have been compared for polyhedral niosomes prepared by hydrating a mixture of a hexadecyl diglycerol ether (C16G2), a poly(24)oxyethylene cholesteryl ether (Solulan C24), 91:9 or 98:2, and conventional spherical niosomes prepared from the same surfactants but with cholesterol. When subjected to osmotic gradients, polyhedral niosomes, the membranes of which are in the gel phase, swell and shrink less than their spherical counterparts and they are more permeable to the hydrophilic solute 5(6) carboxyfluorescein. In 2 M NaCl the rate of release of carboxyfluorescein from polyhedral niosomes (both containing 9% Solulan C24) into either a hypotonic (water) or an isotonic medium (2 M NaCl) was low. This contrasted with similarly loaded spherical niosomes and polyhedral niosomes containing 2% Solulan C24, from which release was high in hypotonic media (e.g. water) but less in an isotonic medium (2 M NaCl). For both polyhedral and spherical niosomes encapsulating carboxyfluorescein (pKa = 6.4), release rates were higher at pH 8 than at pH 5. Polyhedral niosomes are thus, in general, less osmotically active than spherical niosomes because of their rigid but highly permeable membranes. The unusual polyhedral membrane impermeability to carboxyfluorescein co-entrapped with salt in hypotonic media is a function of Solulan C24 content, and is possibly a result of salting out of the polyoxyethylene chains; this is, therefore, a property that might be manipulated in the design of a drug-delivery system. PMID- 10454041 TI - Effect of solvent composition during preparations on the characteristics of enoxacin microparticles. AB - We have studied the effect of the solvent system during preparation on the morphology, encapsulation efficiency, and release characteristics of enoxacin microparticles intended for localized delivery to the bone for the treatment of bone infections. Microparticles of enoxacin were formulated using poly(glycolic acid-co-DL-lactic acid) (PGLA) of different viscosity grades by the solvent evaporation technique. Microparticles prepared with pure dichloromethane had smoother surfaces and less tendency to aggregate than microparticles prepared with dichloromethane-acetone solvent mixtures, which had porous surfaces. Approximately 65% of the microparticles prepared with pure dichloromethane were < 125 microm in diameter compared with 16% (approx.) of microparticles prepared with dichloromethane-acetone mixtures. Increasing the proportion of acetone from dichloromethane-acetone, 10:0, to dichloromethane-acetone, 1:1, resulted in an increase in encapsulation efficiency from 25 to 37%, and an increase in the yield of microparticles harvested from 39 to 51%. Although a further increase in the amount of acetone to dichloromethane-acetone, 1:9, had no significant effect on the yield, aggregation, or fraction of microparticles below 125 microm in diameter, the encapsulation efficiency increased to 56%. Approximately 55% of enoxacin was released in 24 h for microparticles prepared with dichloromethane acetone, 1:9, compared with 100% release in 10h and 2h for microparticles of the same size range prepared with dichloromethane-acetone, 1:1, and dichloromethane acetone, 10:0, respectively. The results suggest that the composition of the dichloromethane-acetone solvent system significantly influences the encapsulation efficiency and the rate of release of enoxacin from microparticles. This is important for the formulation of sustained-release enoxacin microparticles for the localized treatment of osteomyelitis. PMID- 10454042 TI - Evaluation of the factors influencing stomach-specific delivery of antibacterial agents for Helicobacter pylori infection. AB - Because Helicobacter pylori infection is localized in the gastric mucus layer and at the mucus layer-epithelial cell interface, we have developed amoxycillin- and metronidazole-containing chitosan microspheres for stomach-specific drug delivery. Drug-loaded porous chitosan microspheres were prepared by simultaneous crosslinking and precipitation with sodium tripolyphosphate. The release of antibacterial agents into simulated gastric fluid (SGF, pH 1.2), and the stability and permeability through gastric mucin, were examined at 37 degrees C. Because of the high porosity of drug-loaded chitosan microspheres, all the amoxycillin and metronidazole were released in 2 h. High-performance liquid chromatography assays of the antibacterial agents in SGF at 37 degrees C indicated 40% degradation of amoxycillin after 10 h. Metronidazole was completely stable for up to 24 h in SGF. Amoxycillin and metronidazole were highly permeable through the gastric mucin gel layer. The results of this study show that acid stable antibacterial agents, such as metronidazole, that rapidly permeate the gastric mucus layer would be very effective for the complete eradication of H. pylori infection when delivered specifically at the site of infection in the stomach. PMID- 10454043 TI - Comparison of different membranes with cultures of keratinocytes from man for percutaneous absorption of nitroglycerine. AB - The permeability barrier function of cell-culture membranes to the permeation of nitroglycerine was evaluated to find an alternative to skin from man for ex-vivo skin-permeation tests. The membranes were prepared, under submerged conditions, by inducing the growth of keratinocytes, from different donors, on a film of esterified jaluronic acid for different times (10, 20 and 30 days). Their permeability barrier functions were compared with those of some of the most widely used artificial membranes, silicone rubber (Silastic), cellulosic material (Cuprophan, Millipore HAWP), polysulphone membrane (Supor) and polytetrafluoroethylene membrane (TF-PTFE), and with those of biological membranes such as fresh and frozen skin, stratum corneum and epidermis from man, and hairless mouse skin. For each membrane the permeation profile was obtained and the flux was calculated. The permeation profiles for nitroglycerine were similar and linear in the first 2-3 h for all the synthetic membranes tested except TF-PTFE. For this membrane the profile was linear throughout the period considered and the amount permeating in 24 h (1603 microg cm(-2)) was significantly lower than those obtained for the other artificial membranes (between 1926 and 2508 microg cm(-2)). The amounts permeating through all the biological membranes in 24 h were in the range 520 to 781 microg cm(-2), except those for the keratinocyte-culture membranes, which were in the range 1730 to 2553 microg cm(-2). Prolonging the growth period of cultured keratinocytes did not affect nitroglycerine permeation. The findings suggest that these keratinocyte-culture membranes have some advantages--good reproducibility if obtained from the same donor; many membranes can be obtained from the same donor; the preparation is simple; they can be handled more easily than traditional cell culture membranes; and they afford constant penetration rates for a longer period than synthetic membranes. The membranes could be used for preliminary in-vitro permeation studies. PMID- 10454044 TI - Influence of infused catecholamines on the pharmacokinetics of cocaine and benzoylecgonine formation after bolus dose or continuous cocaine administration in the rat. AB - The purpose of this study was to determine whether a catecholamine infusion administered to simulate a stress state could alter the pharmacokinetics of administered cocaine and effect the formation of benzoylecgonine, its major metabolite, in the rat. In a previous investigation we determined that catecholamine infusion enhanced the toxicity of continuous cocaine infusion by reducing the time before the onset of convulsions and respiratory arrest. We postulated that this enhanced toxicity was an effect of catecholamines on the pharmacokinetics of cocaine. To test this hypothesis we studied plasma cocaine and benzoylecgonine disposition after intravenous bolus administration of cocaine (5 mg kg(-1)) to 19 male Sprague-Dawley rats and to 10 rats which received an initial loading-dose cocaine infusion of 1 mg kg(-1) min(-1) (for 5 min) followed by continuous infusion of 100 microg kg(-1) min(-1). Rats in both studies randomly received either continuous catecholamine infusion comprising adrenaline (7.25 microg mL(-1)), noradrenaline (4.4 microg mL(-1)) and dopamine (8.0 microg mL(-1)) or saline, administered at a similar rate. Bolus dose cocaine administration, simultaneously with catecholamine infusion, resulted in significantly higher Cmax levels for cocaine (3.8 compared with 2.5 microg mL( 1)) and lower distribution half-lives (3.3 compared with 5.9 min) and central compartment volumes of distribution (1.5 compared with 2.1 L kg(-1)) compared with saline infusion. Benzoylecgonine formation was significantly reduced in rats receiving catecholamines whereas the elimination half-lives (26.3 compared with 25.0 min) and systemic clearances (146 compared with 146 mL kg(-1) min(-1)) were not different. Continuous cocaine infusion (after an initial loading infusion) resulted in the doubling of plasma cocaine levels in rats receiving catecholamines compared with the control group. These data indicate that elevated plasma catecholamines have significant effects on cocaine pharmacokinetics. This might serve to explain the enhanced toxicity from concomitant cocaine and catecholamine infusion demonstrated in previous experiments. PMID- 10454045 TI - [3H]AL-5848 ([3H]9beta-(+)-Fluprostenol). Carboxylic acid of travoprost (AL 6221), a novel FP prostaglandin to study the pharmacology and autoradiographic localization of the FP receptor. AB - AL-5848 (5Z,13E)-(9 S,11R,15S)-9,11,15-trihydroxy-5,13-prostadienoic acid) is the carboxylic acid of travoprost (AL-6221), a single (+)-isomer of (+/-) fluprostenol, an FP-class prostaglandin agonist which lowers intraocular pressure. We have prepared a radioligand from this selective prostaglandin and demonstrated its utility for studying the pharmacology and autoradiographic location of the FP-receptor. Specific [3H]AL-5848 binding (84% of total) was linearly related to bovine corpus luteum tissue concentration and reached equilibrium within 275 min at 23 degrees C. Scatchard analysis of saturation isotherms indicated interaction of [3H]AL-5848 with a single class of high affinity (dissociation constant, Kd, = 33.8+/-2.9 nM, n = 4) and saturable (Bmax = 37.3+/-3.0 pmol (g wet weight tissue)(-1)) FP receptor-binding sites in bovine corpus luteum. Specific [3H]AL-5848 binding was potently inhibited by the FP receptor ligands 16-phenoxyPGF2alpha (inhibition constant Ki = 17.3 nM); cloprostenol (Ki = 56.8 nM); 17-phenyl PGF2alpha (Ki = 87.0 nM); AL-5848 (Ki = 52.1 nM); PGF2alpha (Ki = 195 nM); PHXA85 (Ki = 223 nM); (n = 3-11) but very weakly by PGD2, ZK118182, BW245C, PGE2, PGI2 and U-46619. The pharmacology of specific [3H]AL-5848 binding correlated well with the pharmacology of [3H]PGF2alpha binding in the bovine corpus luteum preparation (r = 0.98, n = 14, P<0.0001) and also with functional responses in Swiss 3T3 and rat vascular smooth muscle cells (A7r5) (r = 0.96) expressing FP receptors. Autoradiographic studies revealed high levels of specific FP-receptor binding with [3H]AL-5848 on granulosa cells in the bovine corpus luteum sections, and on longitudinal ciliary muscle, the ciliary process, the iris sphincter and the retina in eye sections from man. These studies show [3H]AL-5848 to be a high-affinity agonist radioligand capable of selectively labelling the FP prostaglandin receptor. PMID- 10454046 TI - Thromboxane A2 receptor antagonism in man and rat by a sulphonylcyanoguanidine (BM-144) and a sulphonylurea (BM-500). AB - Torasemide, a loop diuretic, has been reported to relax dog coronary artery precontracted by thromboxane A2 (TXA2), an endogenous prostanoid involved in cardiovascular and pulmonary diseases. N-cyano-N'-[[4-(3'-methylphenylamino)pyrid 3-yl]sulphonyl] homopiperidinoamidine (BM-144) and N-isopropyl-N'-[5-nitro-2-(3' methylphenylamino)-benzenesulphon yl]urea (BM-500), chemically related to torasemide, have been examined for their TXA2 antagonism. The affinity (IC50, the concentration resulting in 50% inhibition) of BM-144 and BM-500 for the TXA2 receptor of washed platelets from man was 0.28 and 0.079 microM, respectively. This is better than for sulotroban (IC50 = 0.93 microM) but less than for SQ 29548 (IC50 = 0.021 microM), two TXA2 antagonists used as reference. The aggregation of platelets from man induced by arachidonic acid was prevented by BM 144 (IC50 = 9.0 microM) and by BM-500 (IC50 = 14.2 microM). Similar results were obtained by use of U-46619, a TXA agonist, as aggregating agent (BM-144, IC50 = 12.9 microM and BM-500, IC50 = 9.9 microM). The contracting effect of U-46619 on rat stomach strip was abolished by BM-144 (IC50 = 1.01 microM) and BM-500 (IC50 = 2.54 microM). Both drugs (BM-144: IC50 = 0.12 microM and BM-500: IC50 = 0.19 microM) also relaxed rat aorta precontracted by U-46619; both were more potent than sulotroban (IC50 = 1.62 microM). The two torasemide derivatives (100 microM) did not significantly affect the myo-stimulating effect of some prostaglandins (PGE2, PGI2, PGF2alpha) or aorta contraction elicited by KCl (30 mM). They did not modify rat diuresis after administration of a 30-mg kg(-1) dose. In conclusion, BM-144 and BM-500 can be regarded as novel non-carboxylic TXA2 receptor antagonists and offer a novel template for the design of more potent molecules. PMID- 10454047 TI - Anti-inflammatory profile of N-phenylpyrazole arylhydrazone derivatives in rats. AB - A series of synthetic N-phenylpyrazole arylhydrazone compounds, rationally designed as mixed-hybrid isosteres of two known inhibitors of prostaglandin synthase and 5-lipoxygenase enzymes, BW-755c and CBS-1108, has been investigated for anti-inflammatory activity in the carrageenan-induced pleurisy model in rats. The compounds have different oxygenated substituent groups in the aryl group of the hydrazone framework to ensure a different range of redox properties. A new arylhydrazone derivative, 2,6-di-tert-butyl-4-(4-nitro-3-methyl-N-phenylpyrazol-5 yl-hydr azonomethyl)phenol, was also synthesized and tested for anti-inflammatory activity. Although all the compounds significantly inhibited (by 30-90%) neutrophil accumulation in the pleural cavity, there was great variability in the anti-oedematogenic effect of the compounds (3-96%). 5-(4'-Hydroxy-3' methoxybenzylidene)hydrazone-3-methyl-4-nitrop henylpyrazole was the most active compound in this series; it had a remarkable antiinflammatory profile, almost blocking both assays. In contrast, the compound with a 2,6-di-tert-butylated hydroxybenzene ring on the hydrazone group inhibited neutrophil migration only. These results will be useful for further structure-activity relationship studies devoted to improving the dual prostaglandin synthase-5-lipoxygenase activity of these derivatives and determining the minimum structural requirements necessary for this activity. PMID- 10454048 TI - Vascular-selective effect of lercanidipine and other 1,4-dihydropyridines in isolated rabbit tissues. AB - The aim of this study was to characterize the in-vitro vasoselectivity of lercanidipine (in comparison with lacidipine, amlodipine, nitrendipine and felodipine) by evaluating its functional calcium antagonistic activity on rabbit vascular (aorta) and cardiac tissues (heart ventricle). Although incubation with all the compounds tested elicited a concentration-dependent relaxant effect on vascular tissue precontracted with KCl (80 mM), 50% relaxation was reached at different times for each concentration and drug tested. At 10 nM concentration 50% relaxation was reached after 210 min with lercanidipine, 278 min with amlodipine, 135 min with lacidipine, 75 min with nitrendipine and 70 min with felodipine. The onset of the effect was, therefore, similar for lercanidipine, amlodipine and lacidipine, but faster for nitrendipine and felodipine. Similarly, all the compounds tested concentration-dependently reduced the force of cardiac contraction (negative inotropic activity). In this model, the time needed to reach 50% reduction in contractile force was also concentration-dependent, and the ranking order of the speed of onset of the effect (evaluated as the ratio of the IC50 values (the concentrations inhibiting contraction by 50%) calculated after 1 and 4 h incubation) was lacidipine (3.8) > amlodipine (9.6) > felodipine (39) > lercanidipine (68) = nitrendipine (89). The vasoselectivity, expressed as the ratio of the IC50 values obtained on cardiac and vascular tissue, were (for 4 h incubation) 730, 193, 95, 6 and 3 for lercanidipine, lacidipine, amlodipine, felodipine and nitrendipine, respectively, showing that lercanidipine is the most vasoselective of the calcium-antagonists tested. The results show that lercanidipine reduces the inotropic force of the rabbit heart to a lesser extent than do other calcium antagonists, and that this drug had the best heart/vessel selectivity index among the compounds tested at all the times tested. PMID- 10454049 TI - Evaluation of the protective effects of alkaloids isolated from the hooks and stems of Uncaria sinensis on glutamate-induced neuronal death in cultured cerebellar granule cells from rats. AB - We have previously shown that an aqueous extract of the hooks and stems of Uncaria sinensis (Oliv.) Havil., Uncariae Uncus Cum Ramulusis, protects against glutamate-induced neuronal death in cultured cerebellar granule cells by inhibition of Ca2+ influx. Because it is not known which components of Uncaria sinensis are active, in this study we have evaluated, by 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyl-tetrazolium bromide (MTT) staining, the neuroprotective effects of the oxyindole alkaloids corynoxeine, rhynchophylline, isorhynchophylline and isocorynoxeine, and the indole alkaloids geissoschizine methyl ether, hirsuteine and hirsutine, isolated from the hooks and stems of Uncaria sinensis, on glutamate-induced cell death. We also investigated the inhibitory effects of the compounds on 45Ca2+ influx in cultured rat cerebellar granule cells. Cell viability evaluated by the MTT assay was significantly increased by application of rhynchophylline (10(-3) M), isorhynchophylline (10(-4)-10(-3) M), isocorynoxeine (10(-4)-10(-3) M), hirsuteine (10(-4)-3 x 10(-4) M) or hirsutine (10(-4)-3 x 10(-4) M) compared with exposure to glutamate only, with the effect of isorhynchophylline being the strongest. The increased 45Ca2+ influx into cells induced by glutamate was significantly inhibited by administration of rhynchophylline (10(-3) M), isorhynchophylline (3 x 10(-4)-10(-3) M), isocorynoxeine (3 x 10(-4)-10(-3) M), geissoschizine methyl ether (10(-3) M), hirsuteine (3 x 10(-4)-10(-3) M) or hirsutine (3 x 10(-4)-10(-3) M). These results suggest that oxyindole alkaloids such as isorhynchophylline, isocorynoxeine and rhynchophylline and indole alkaloids such as hirsuteine and hirsutine are the active components of the hooks and stems of Uncaria sinensis which protect against glutamate-induced neuronal death in cultured cerebellar granule cells by inhibition of Ca2+ influx. PMID- 10454051 TI - Hepatoprotective properties in the rat of Mitracarpus scaber (Rubiaceae). AB - The effect of Mitracarpus scaber on carbon tetrachloride-induced acute liver damage in the rat has been evaluated. Results showed that treatment with Mitracarpus scaber decoction resulted in significant hepatoprotection against CCl4-induced liver injury both in-vivo and in-vitro. In-vivo, Mitracarpus scaber pretreatment reduced levels of serum glutamate-oxalate-transaminase (P < 0.01 for 250, 500 and 1000 mg kg(-1)) and serum glutamate-pyruvate-transaminase (P < 0.05 for 250 mg kg(-1) and P < 0.01 for 1000 mg kg(-1)) previously increased by administration of CCl4. In-vitro results indicated that addition to the culture medium of Mitracarpus scaber extracts significantly reduced glutamate-oxalate transaminase (P < 0.05 for 100 microg mL(-1) and P < 0.01 for 10 and 1000 microg mL(-1)) and lactate dehydrogenase activity (P < 0.05 for 10 microg mL(-1)). Mitracarpus treatment also resulted in a good ( > 93%) survival rate for the CCl4 intoxicated hepatocytes as demonstrated by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay. Moreover, as in the in-vitro assay, Mitracarpus scaber had radical-scavenging properties, shown by its reaction with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (EC50, the extract concentration resulting in a 50% reduction in the absorbance of DPPH blank solution, = 41.64+/ 1.5 microg mL(-1)). The results of this study showed that Mitracarpus scaber had antihepatotoxic potential, a finding which supports the validity of traditional usage of this drug in Mali for the treatment of liver diseases. PMID- 10454050 TI - Effects of Hypericum perforatum on levels of 5-hydroxytryptamine, noradrenaline and dopamine in the cortex, diencephalon and brainstem of the rat. AB - The plant Hypericum perforatum is used in folk medicine to treat several diseases and research attention has been recently focused on its antidepressant action. Hypericin and flavonoids are the most important constituents of the plant, but the exact role of these compounds in the effects of hypericum on mood disorders is not well known. We have investigated the contribution of these compounds to the antidepressant effects of hypericum. The effects of acute administration of hypericum extracts on levels of 5-hydroxytryptamine (5-HT), tryptophan, 5 hydroxyindoleacetic acid (5-HIAA), noradrenaline and dopamine in the cortex, diencephalon and brainstem was evaluated. The levels of these neurotransmitters were measured 1 h and 24 h after administration of two different extracts, one containing 0.3% hypericin and 6% flavonoids (Li 160; 25-500 mgkg(-1)), the other containing 0.3% hypericin and 50% flavonoids (Ph-50; 25-500 mgkg(-1)). Results from experiments performed on 5-HT turnover were compared with the effects of fluoxetine (10-80 mgkg(-1)). Li 160, Ph-50 and fluoxetine induced a significant increase in the 5-HT content of the cortex. In the diencephalon Ph-50, but not Li 160 or fluoxetine, elicited an increase in 5-HT and 5-HIAA levels. In the brainstem Ph-50 and fluoxetine caused an increase in 5-HT content; Li 160 did not change neurotransmitter content. Both Li 160 and Ph-50 caused increases of noradrenaline and dopamine in the diencephalon. In the brainstem only Ph-50 induced an increase in noradrenaline content. Our data confirm that acute administration of hypericum extracts modifies the levels of neurotransmitters involved in the pathophysiology of mood disorders. When the extracts contain a higher concentration of flavonoids the effects are more widespread and involve brain regions such as diencephalon and brainstem that are implicated in depression. PMID- 10454052 TI - Anti-ulcerogenic mechanisms of a lyophilized aqueous extract of Dalbergia monetaria L. in rats, mice and guinea-pigs. AB - The decoction of Dalbergia monetaria L. is popularly used in Brazil for the treatment of gastric ulcer and the lyophilized aqueous extract (LAE) of D. monetaria has significant anti-ulcerogenic activity and inhibits gastric ulcer lesions induced by pylorus-ligature, ethanol and hypothermic-restraint stress. This work was conducted to identify the antiulcerogenic mechanisms of action of the LAE of D. monetaria. We analysed the effect of the LAE on prostaglandin E2 (PGE2) synthesis and on the characteristics (pH, volume and total acid content) of gastric juice. The antagonism between the LAE and histamine or carbachol was also analysed. The LAE increased gastric mucosal PGE2 synthesis compared with control (89.7+/-21.5 and 52.6+/-11.8 pg mg(-1), respectively) as assayed by enzyme immunoassay in the rat stomach. The LAE reduced the total acid content of gastric juice, but did not modify pH or gastric volume significantly, in Shay rats. Dose-response curves to histamine were shifted to the right in guinea-pig isolated right atria (pD2 values were 5.77+/-0.2 and 5.42+/-0.3, respectively, in the absence and presence of the LAE), with a significant reduction in maximum response (140+/-15.1 and 98+/-13.0, respectively). The same effect was observed when the agonist was isoprenaline. The LAE had no effect on the dose-response curve to carbachol in rat fundus strips. Thus, the protective effect of the LAE on induced gastric lesions might be because of synergistic effects, e.g. increased PGE2 synthesis and antagonism of H2 histamine and beta-adrenergic receptors, reducing gastric acid secretion. Increased PGE2 synthesis results in increased protection, and antagonism of H2 histamine and beta-adrenergic receptors reduces aggressive factors against the gastric mucosa. PMID- 10454053 TI - Effect of some antidepressants on glycaemia and insulin levels of normoglycaemic and alloxan-induced hyperglycaemic mice. AB - Depression is an important problem among diabetic patients. We have investigated the effect of some antidepressant drugs on plasma glucose and insulin levels in normoglycaemic and alloxan-induced diabetic mice. For this purpose the effects of nortryptiline (as an example of a tricyclic antidepressant) and fluoxetine and sertraline (as examples of selective 5-HT re-uptake inhibitors) were examined on plasma glucose and insulin levels. Nortryptiline significantly increased glucose levels and reduced insulin levels in all animals. Although neither fluoxetine nor sertraline induced changes in insulin levels, both significantly reduced the blood glucose levels of mice. These results suggest that antidepressive treatment has important risks particularly for diabetics. Tricyclic antidepressants might induce an important decrease in glucose tolerance and worsen the control of diabetic patients. Selective 5-HT re-uptake inhibitors, on the other hand, might reduce plasma glucose independently of insulin levels. This point is particularly important and should be remembered when insulin or oral antidiabetic agents are administered to diabetics, because of the possible risk of hypoglycaemia. PMID- 10454054 TI - Influence of dietary iodine on drug-induced hypothyrodism in the rat. AB - Several compounds of pharmaceutical importance from a variety of chemical families, for example chlorpromazine and clomipramine, have been found to form charge-transfer complexes with iodine. We have investigated the influence of dietary iodine on thyroid-gland dysfunction induced by clomipramine, chlorpromazine or 2-thiazoline-2-thiol. We suggest that iodine is partly diverted from its metabolic pathway by complexation with drugs, and so the urinary concentration of iodide is increased. Both chlorpromazine and clomipramine, at doses which do not inhibit thyroperoxidase, enhanced urinary iodine excretion when dietary iodine was restricted (3.944+/-0.96 microg/day for chlorpromazine tested rats, 3.43+/-1.33 microg/day for clomipramine-tested rats, compared with 2.34+/-0.11 microg/day in control rats). Concurrently, these pharmaceutical compounds increased the level of free thyroid-stimulating hormone (TSH) in comparison with controls and induced histological modifications in, and enlargement of, the thyroid gland. We have demonstrated that drug-induced loss of iodine in the urine was associated with antithyroid action when iodine intake was limited. PMID- 10454055 TI - Brittle-ductile transitions in organic solids during comminution: a practical demonstration. PMID- 10454056 TI - Non-invasive assessment of autonomic nervous system integrity in able-bodied and spinal cord-injured individuals. AB - The purpose of this study is to identify features of the Valsalva maneuver (VM) that differentiate between able-bodied (AB, n = 14) and spinal cord-injured individuals with high (T, n = 11) and low (P, n = 10) lesions. The systolic blood pressure (BP) recovery during the VM was used as a measure of sympathetic and vagal activity, whereas from the BP rise in phase IV of the VM baroreflex sensitivity was derived. After a similar initial BP decrease in all groups, BP recovered faster and more completely in AB compared to P and T during the VM. After release of the VM, the BP overshoot was very similar in AB and P, suggesting that the BP overshoot reflects sympathetic activity onto the heart. The baroreflex sensitivity was smaller in P, but not in T, compared to AB. However, 36% of the observations of T had such a low correlation between BP and RR interval that a slope could not be determined, which caused loss of observations, but suggested a low baroreflex sensitivity as well. PMID- 10454057 TI - The effects of exposure to moderate altitude on cardiovascular autonomic function in normal subjects. AB - Cardiovascular responses to altitude have been studied on well-trained young subjects, generally at high altitudes (>4000 m). Less known are the effects of exposure to lower altitudes, easily reached by the general population. The aim of the study was to evaluate the effects of exposure to a moderate altitude (2950 m) on heart rate (HR), blood pressure (BP) profile, and cardiovascular autonomic function, and their correlation with hemoglobin oxygen saturation (HbO2S), in untrained subjects of a wide age range. Twenty-seven healthy normotensive subjects (age range 6-83; 8 children, 9 adults, and 10 elderly subjects) underwent a battery of noninvasive cardiovascular reflex tests and 24-h ambulatory BP monitoring. Corrected QT interval was also calculated. HbO2S was measured with a transcutaneous oxymeter. All measurements were performed at about 200 m (s.l.) and repeated at 2950 m. 24-h HR and systolic/diastolic BP mean values increased at 2950 m in children (% change respectively: 6.4 +/- 6.4, p<0.05; 6.5 +/- 4.0/13.5 +/- 6.9, p < 0.05), adults (4.9 +/- 8.1, NS; 6.0 +/- 5.1/8.1 +/- 5.8, p < 0.05), and elderly subjects (7.2 +/- 4.8, p < 0.05; 5.1 +/- 2.3/2.8 +/- 4.1, p < 0.05 for systolic BP only). Standard deviation of BP mean values increased during night-time in the adult group (p < 0.05). All subjects scored normal cardiovascular test results and no differences were observed after exposure to 2950m, at both 1 hour and 24 hours from arrival. After exposure to altitude, HbO2S decreased significantly in the three groups, both on arrival and after 24 hours. No correlation was found between changes in HbO2S and BP/HR responses, and cardiovascular test results. In conclusion, exposure to moderate altitudes, easily and often reached by the general population, causes a small but significant increase in BP and HR in healthy untrained subjects of a wide age range (6-83 years). Some physiological factors (eg, lower environmental temperature and lifestyle modification) together with hypoxia, possibly more than altered cardiovascular reactivity, seem responsible for this cardiovascular change. In terms of end-organ damage, the clinical relevance of this increase in BP and BP variability for repeated exposure is not known. PMID- 10454058 TI - Exercise test in Parkinson's disease. AB - In this study we assessed cardiovascular performance and metabolic response after an exercise test in Parkinsonian patients (PD patients). METHODS: 15 PD patients (10 male, 5 female; mean age:63 +/- 6.17 y; mean weight: 72.2 +/- 9.5 kg) and 15 sex, age (mean: 63.8 +/- 5.38 y), and weight-(mean: 72.2 +/- 8.69 kg) matched controls performed an exercise test using a cycle ergometer and a ramp protocol. All patients and control subjects underwent a heart rate variability test prior to the exercise test. At rest and at the end of each interval blood pressure (BP) and capillary lactate samples were taken. Heart rate was monitored continuously. RESULTS: The heart rate variability tests were abnormal in Parkinson's patients. All patients and control subjects achieved an intensity level of 75 watts, 12/15 PD patients managed 100 watts, and 7/15 PD patients managed 150 watts. 12/15 control subjects performed at an intensity level of 125 Watts and 9/15 at an intensity level of 150 Watts. There was no statistically significant difference in heart rate increase but there was a tendency to lower lactate levels at high intensity levels in PD patients. PD patients had a statistically lower systolic BP at 75, 100, 125, and 150 watts. CONCLUSION: We did not find striking differences in cardiovascular adaptation to physical work in PD patients and we propose, therefore, that it should be possible to improve cardiovascular endurance in PD patients. Previously, reports have suggested respiratory chain impairment in Parkinson's disease and poor endurance performance. However, our results do not support a clinically relevant impairment of the respiratory chain. PMID- 10454059 TI - A case of sinus arrest and vagal overactivity during REM sleep. AB - A young man presented with tachycardia and faintness after an episode of influenza. He underwent 24-h heart rate recordings, each of which documented episodes of sinus arrest lasting up to 7.2 seconds. All episodes occurred in the second half of the night and were always accompanied by severe bradycardia. Cardiac function tests failed to disclose anything abnormal. Two polysomnographic recordings demonstrated that the sinus arrests occurred during REM sleep. Power spectral analysis of heart rate variability showed that during the second half of the night there was an abnormal prevalence of vagal activity, particularly during REM sleep stages, presumably responsible for the bradycardia and fall in blood pressure. We speculate that the episodes of sinus arrest are linked to a central mechanism that triggers the autonomic imbalance during REM sleep. PMID- 10454060 TI - Clinical and physiological characteristics of autonomic failure with Parkinson's disease. AB - We analyzed the clinical and physiological features of autonomic failure with Parkinson's disease (AF-PD) in seven patients and compared them with those of autonomic failure with multiple system atrophy (AF-MSA). In AF-PD, parkinsonism was more gradually progressive than in AF-MSA, and symptoms were responsive to L dopa. All seven patients with AF-PD had orthostatic hypotension, postprandial hypotension, and constipation, but no urinary retention. Of these, three had hypohidrosis and five had frequent urination; five patients had subnormal plasma norepinephrine (NE) concentrations. Supersensitivity to NE infusion was observed in all patients. Head-up tilting (HUT) test resulted in no increase of plasma NE concentrations in both groups, but a significant increase of the plasma arginine vasopressin (AVP) concentrations in the patients with AF-PD. Urodynamic studies revealed that urinary bladder function was relatively well preserved in AF-PD in contrast to AF-MSA. In conclusion, there exists some clinical and physiological differences in autonomic features between AF-PD and AF-MSA, and postganglionic involvement predominates in AF-PD. PMID- 10454061 TI - One hundred years of adrenaline: the discovery of autoreceptors. AB - The active principle of suprenal extract that produces its pressor effects was isolated by the joint research of John Abel in 1899 and Jokichi Takamine in 1901. Within three years Elliott, working in Langley's laboratory, suggested that this active principle, referred to by British physiologists as "adrenaline" and named "Adrenalin" by Takamine, was released from sympathetic nerve terminals to act on smooth muscle cells. However, it was not until 1946 that von Euler showed that demythelated adrenaline (noradrenaline) rather than adrenaline is a sympathetic transmitter. The possibility that this sympathetic transmitter could also act on nerve terminals was not developed until 1971. Research on autoreceptors culminated in the identification of adrenergic receptors on nerve terminals different to those on muscle cells. This paper assesses the contributions that established the idea of the adrenergic autoreceptor, 100 years after the discovery of adrenaline. PMID- 10454062 TI - Harvard Report on Cancer Prevention. Volume 3: prevention of colon cancer in the United States. PMID- 10454063 TI - Incidence of thyroid cancer in Scandinavia following fallout from atomic bomb testing: an analysis of birth cohorts. AB - OBJECTIVES: The occurrence relation between radioactive fallout from nuclear testing at Novaja Semlja in north-west Russia and the incidence of thyroid cancer in Norway and Sweden was studied following a birth cohort approach. METHODS: Birth cohorts with presumably different levels of exposure were identified according to calendar year of atomic tests and previous Norwegian estimates of the population dose (born 1947-1950 received low exposure in late childhood, born 1951-1962 received the highest exposure in early childhood, born 1963-1970 were not exposed). For each one-year birth cohort the incidence rates were calculated, with denominators based on exact population figures for each year of follow-up. RESULTS: In a stratified analysis, the relative risk for the highest exposed cohorts born 1951-1962, compared to those not exposed born 1963-1970, was found to decrease with increasing age from a borderline significant relative risk (RR) of 1.7 (95 percent confidence interval, 95% CI: 1.0-3.0) for children in the age group 7-14 years to no excess risk among those 20-24 years of age (RR: 0.9; 95% CI: 0.7-1.2). The mean age at diagnosis of thyroid cancer in the age-group 7 14 years was lowest in the birth cohorts with the highest exposure. The Poisson regression analysis showed essentially the same results, with an improved fit when adding an interaction term between age and birth-cohort to a basic model with age, gender, birth-cohort and country. CONCLUSION: These results are compatible with an increased risk of thyroid cancer during childhood and adolescence for subjects exposed to radioactive fallout early in life. Alternative explanations for the pattern of incidence are discussed. PMID- 10454064 TI - Diet in adolescence and the risk of breast cancer: results of the Netherlands Cohort Study. AB - OBJECTIVES: In The Netherlands, part of the population experienced food restriction and severe famine during World War II. The purpose of this study was to study the effects of severe undernutrition during adolescence on the risk of breast cancer later in life. METHODS: We examined the hypothesis in the Netherlands Cohort Study on diet and cancer (NLCS), among 62,573 women aged 55-69 years. Baseline information on diet and other risk factors was collected with a questionnaire in 1986. Information was collected on residence in the Hunger winter (1944-1945) and War years (1940-1944) and fathers' employment status in 1932-1940 as indicators of exposure. After 6.3 years of follow-up, 1009 incident breast cases were available for analysis. RESULTS: In multivariate case-cohort analysis, residents of the western part of the country in 1944-1945 had an increased breast cancer risk (western city RR = 1.1, 95% CI: 0.9-1.4, western rural area RR = 1.5, 95% CI: 1.1-1.9). For the War years (1940-1944) we found no association between breast cancer risk and urban vs. rural residence. Women whose fathers were unemployed during the Depression years (1932-1940) had a non significant decrease in breast cancer risk (RR = 0.9, 95% CI: 0.7-1.2). Exposure to energy restriction during the adolescent growth spurt or during the period between menarche and birth of the first child did not change the RRs substantially. CONCLUSIONS: We found no clear evidence in this study for the hypothesis that energy restriction in adolescence leads to a decreased breast cancer risk. PMID- 10454065 TI - Cigarette smoking, alcohol consumption and risk of nasopharyngeal carcinoma in Taiwan. AB - OBJECTIVES: Nasopharyngeal carcinoma (NPC) is rare in most countries but occurs with relatively high frequency among southern Chinese populations throughout the world. A case-control study of NPC was conducted in Taiwan to investigate the importance of active and passive cigarette exposure and alcohol consumption as risk factors for this disease. METHODS: 375 histologically confirmed incident NPC cases (99% response rate) were prospectively identified from two hospitals in Taipei between July 1991 and December 1994 and administered a detailed questionnaire. 327 healthy community controls individually matched to cases on sex, age and residence were selected (88% response rate). RESULTS: After multivariate adjustment, the odds ratio (OR) and 95% confidence interval (CI) was 1.7 (1.1-2.9 with p = 0.03 for increasing dose-response) for those who smoked for > or = 25 years compared with non-smokers. Passive smoking during childhood or adult life was not associated with an increased risk of disease. Alcohol consumption was not associated with NPC risk. The OR for subjects with > or = 15 grams of ethanol per day compared to non-drinkers was 1.1 (95% CI = 0.7-1.7). CONCLUSIONS: Our results suggest that long term cigarette smoking is associated with NPC but that low level exposure to cigarette smoke via passive exposure and alcohol consumption are not associated with disease risk. PMID- 10454066 TI - Occupation and bladder cancer in European women. AB - OBJECTIVES: We examined the importance of occupational exposures for bladder cancer in women. METHODS: We combined data from 11 case-control studies conducted between 1976 and 1996 in six European countries. The pooled data comprised 700 incident female cases and 2425 population or hospital controls, aged 30-79 years. Lifetime occupational and smoking history were examined using common coding. RESULTS: Excess risks were found in only a few of the occupations previously identified at high risk for bladder cancer. Statistically significant excess risks were observed for metal workers, particularly blacksmiths, toolmakers and machine tool operators (OR: 2.0, 95% CI: 1.1-3.6), tobacco workers (OR: 3.1, 95% CI: 1.1-9.3), field crop and vegetable farm workers (OR: 1.8, 95% CI: 1.0-3.1), tailors and dress makers (OR: 1.4, 95% CI: 1.0-2.1), saleswomen (OR: 2.6, 95% CI: 1.0-6.9), and mail sorting clerks (OR: 4.4, 95% CI: 1.0-19.5). About 8% (95% CI: 3.1 19.9) of all bladder cancers in women could be attributed to occupation after adjusting for smoking. The attributable risk was higher in women aged less than 65 years (12%), compared to older women (4%). CONCLUSIONS: The calculation of the attributable risk on the basis of results from this analysis may have caused some overestimation of the proportion of occupational bladder cancer in women. A significant proportion, however, of bladder cancer cases among European women less than 65 years is likely to be attributed to occupation. This link between bladder cancer in women and occupational factors has received little recognition, probably because studies addressing these issues have predominantly been done in men. PMID- 10454067 TI - Association between alcohol and lung cancer in the alpha-tocopherol, beta carotene cancer prevention study in Finland. AB - OBJECTIVES: We evaluated the association between alcohol intake and lung cancer in a trial-based cohort in Finland, the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study (ATBC Study). METHODS: During an average of 7.7 years of follow up, 1059 lung cancer cases were diagnosed among the 27,111 male smokers with complete alcohol and dietary information. The relationship between alcohol and lung cancer was assessed in multivariate Cox regression models that adjusted for age, smoking, body mass index and intervention group. RESULTS: Nondrinkers, 11% of the study population, were at increased lung cancer risk compared to drinkers (RR = 1.2, 95% CI: 1.0-1.4), possibly due to the inclusion of ex-drinkers who had stopped drinking for health reasons. Among drinkers only, we observed no association between lung cancer and total ethanol or specific beverage (beer, wine, spirits) intake. We found no significant effect modification by level of smoking, dietary micronutrients or trial intervention group; however, for men in the highest quartile of alcohol intake, we observed a slight increase in risk for lighter smokers (<1 pack/day) and reduced risk among the heaviest smokers (>30 cigarettes/day). CONCLUSIONS: We concluded that alcohol consumption was not a risk factor for lung cancer among male cigarette smokers, and its effect was not significantly modified by other factors, notably smoking history. PMID- 10454068 TI - The relation of p53 gene mutations to gastric cancer subsite and phenotype. AB - OBJECTIVES: We investigated p53 gene mutations in advanced gastric cancers by direct DNA sequencing, in order to determine the frequency of mutations in gastric cancers having different epidemiological backgrounds, tumors of the cardia were compared with those arising in the antrum or corpus. Intestinal type cancers were compared with diffuse or other histologic types. We have chosen to assess the frequency of mutations solely based on DNA sequencing. METHODS: Paraffin embedded tissues from 100 gastric cancers were evaluated. The mutational status of the p53 gene in exons 5 through 9 were determined by direct sequencing of PCR products. RESULTS: Mutations in exons 5, 6, 7 and 8 were found in 35 of 100(35%)stomach cancers. One tumor had mutations in both exons 5 and 8. No mutations were detected in exon 9. p53 gene mutations were significantly more frequent in cancers of the cardia (19/35; 54%) than the antrum and corpus (16/65 (25%)) (p < or = 0.005). p53 mutations were more frequent in intestinal type cancers (28/67; 42%) than diffuse cancers or other histologic types of cancer (7/33; 21%), but the difference was not statistically significant. CONCLUSIONS: Cancers of the cardia more frequently contain p53 mutations than do antral and corpus cancers, suggesting that cancers in the proximal and distal stomach evolve through different molecular pathways. PMID- 10454069 TI - Childhood leukemia and personal monitoring of residential exposures to electric and magnetic fields in Ontario, Canada. AB - OBJECTIVES: To evaluate the risk of childhood leukemia in relation to residential electric and magnetic field (EMF) exposures. METHODS: A case control study based on 88 cases and 133 controls used different assessment methods to determine EMF exposure in the child's current residence. Cases comprised incident leukemias diagnosed at 0-14 years of age between 1985-1993 from a larger study in southern Ontario; population controls were individually matched to the cases by age and sex. Exposure was measured by a personal monitoring device worn by the child during usual activities at home, by point-in-time measurements in three rooms and according to wire code assigned to the child's residence. RESULTS: An association between magnetic field exposures as measured with the personal monitor and increased risk of leukemia was observed. The risk was more pronounced for those children diagnosed at less than 6 years of age and those with acute lymphoblastic leukemia. Risk estimates associated with magnetic fields tended to increase after adjusting for power consumption and potential confounders with significant odds ratios (OR) (OR: 4.5, 95% confidence interval (CI): 1.3-15.9) observed for exposures > or = 0.14 microTesla (microT). For the most part point-in-time measurements of magnetic fields were associated with non-significant elevations in risk which were generally compatible with previous research. Residential proximity to power lines having a high current configuration was not associated with increased risk of leukemia. Exposures to electric fields as measured by personal monitoring were associated with a decreased leukemia risk. CONCLUSIONS: The findings relating to magnetic field exposures directly measured by personal monitoring support an association with the risk of childhood leukemia. As exposure assessment is refined, the possible role of magnetic fields in the etiology of childhood leukemia becomes more evident. PMID- 10454070 TI - Research of the adaptive response induced by low-dose radiation: where have we been and where should we go? PMID- 10454071 TI - The role of adaptive responses following exposure to ionizing radiation. PMID- 10454072 TI - Radioadaptive response: responses to the five questions. PMID- 10454073 TI - Radiation exposure and adaptive processes. PMID- 10454074 TI - Adaptive response and its relationship to hormesis and low dose cancer risk estimation. PMID- 10454075 TI - The adaptive response to ionizing radiation: low dose effects unpredictable from high dose experiments. PMID- 10454076 TI - Perspectives on the adaptive response from studies on the response to low radiation doses (or to cisplatin) in mammalian cells. PMID- 10454077 TI - Adaptive response induced by low levels of radiation. Summary and comments. PMID- 10454079 TI - Stimulation of survival capacity in heat shocked cells by subsequent exposure to minute amounts of chemical stressors; role of similarity in hsp-inducing effects. AB - A brief and moderate heat shock to Reuber H35 hepatoma cells causes a rapid increase in the synthesis of heat shock proteins (hsp) and initiates the development of thermotolerance, which results in an increased ability to survive exposure to otherwise lethal temperatures. We now demonstrate that low doses of various chemical stressors (arsenite, cadmium, mercury, lead, copper, menadione and diethyldithiocarbamate (ddtc)), at concentrations that do not exert any effect in control cultures, are able to enhance the synthesis of hsps and to stimulate the development of thermotolerance when applied to cultures which were pretreated with a mild heat shock. The degree of stimulation appears to be stressor-specific, which is not only observed in the ensuing development of thermotolerance but also in the enhancement of the heat shock-induced synthesis of stress proteins. The different hsps that show an enhanced induction when heat shocked cultures are exposed to the various secondary applied low doses of chemical stressors, were found to resemble the hsp pattern that is characteristic for the secondary stressor and not for the initial heat shock. In other words, the nature of the post-treatment determines the observed pattern of enhanced synthesis of hsps. In order to analyze the origin of the stimulation of survival capacity by low doses of the mentioned stressors, we studied whether the degree of stimulation is determined by the degree of similarity between the overall stress response to heat shock and to the second stress condition when applied singly. The degree in which low doses of chemical stressors stimulate tolerance development and enhance the synthesis of hsps in cells that were previously heat shocked, appears to be related to the degree of similarity in the hsp pattern induced by both stressors. Our results support the notion that low doses of toxic compounds may, under certain conditions, have beneficial effects related to a stimulation of endogenous cytoprotective mechanisms. PMID- 10454078 TI - Hyperlipidaemia in cisplatin-induced nephrotic rats. AB - The serum and hepatic lipid concentrations were investigated in rats made nephrotic with a single intraperitoneal injection of cisplatin (6 mg kg(-1) b.wt.). The serum creatinine and urea concentrations were estimated as indices of nephrotoxicity, and the serum total bilirubin level as a liver function test. 3 The fasting serum total cholesterol, triglycerides (TG) and the cholesterol fractions associated with the various lipoproteins, as well as hepatic cholesterol and TG contents were also measured, following 5, 10 and 15 days from the cisplatin treatment. 4 The results revealed that on day 5 both serum creatinine and urea concentrations were significantly (P<0.01) increased, indicating the peak of nephrotoxicity, with no injurious effects on the liver as indicated by the unaltered serum bilirubin concentration. 5 The nephrotoxicity was accompanied by significant elevations in serum total cholesterol and TG concentrations by 49 and 42%, respectively, with significant (P < 0.05) correlations between the serum cholesterol and TG concentrations versus the serum urea (r=0.68 and r=0.60, respectively). Among the estimated lipoproteins, very low density lipoprotein (VLDL) cholesterol was severely increased to more than twofold with no severe changes in LDL- or HDL-cholesterol fractions. On day 5 the liver also showed significant accumulation of TG with no change in the cholesterol content. Animals killed 10 or 15 days post-cisplatin treatment had all the perturbed parameters returned to the normal levels. The present results indicated that rats exposed to a single cisplatin injection exhibit acute reversible nephrosis on day 5 which was accompanied by dyslipidaemia and accumulated liver TG. PMID- 10454080 TI - Acute overdose due to benzydamine. AB - Benzydamine is a nonsteroidal anti-inflammatory drug, currently available as mouthwash, aerosol, dermal cream, vaginal douche preparation, pills and otic drops. Up to now no cases of poisoning due to this drug have been reported. A 6 year-old girl with an accidental poisoning with benzydamine is described. The episode consisted of hallucinosis without other symptoms and resolved spontaneously in 17 h. PMID- 10454081 TI - Detection of cortisol administration in the horse. PMID- 10454082 TI - Testicular neoplasia of horses: an underreported condition. PMID- 10454083 TI - Hydrocortisone levels in the urine and blood of horses treated with ACTH. AB - An investigation was undertaken to demonstrate whether therapeutic treatment with ACTH raises hydrocortisone (cortisol) levels in horse urine above the limit (1000 ng/ml) established by the International Conference of Racing Authorities with the aim of controlling the abuse of cortisol and ACTH in equine sports. ACTH (200 iu) was administered i.m. to 3 Thoroughbred horses; urine and blood samples were collected at intervals afterwards and analysed by an immunoenzymatic system (ELISA) and HPLC-MS. To ascertain post exercise cortisol levels in untreated horses, 101 urine and 103 serum samples were taken from horses immediately after racing and analysed by ELISA. The peak urine level of cortisol, detected 8 h after ACTH administration, was around 600 ng/ml using either ELISA or HPLC-MS. The peak serum cortisol concentration was found to be around 250 ng/ml by ELISA, but consistently less by HPLC-MS. Mean cortisol levels in post race horses were 135.1+/-72.1 ng/ml in urine and 90.1+/-41.7 ng/ml in serum. High levels of the metabolite 20beta-dihydrocortisol in urine and the cortisol precursor 11beta desoxycortisol in serum were found. The latter showed high cross-reactivity with cortisol on ELISA. In our experiment, treatment with ACTH 200 iu i.m. did not raise urinary cortisol levels above the 1000 ng/ml threshold proposed by the ICRA. PMID- 10454085 TI - PGFM response to exogenous oxytocin and determination of the half-life of oxytocin in nonpregnant mares. AB - We investigated the half-life of oxytocin in reproductively normal mares and the prostaglandin response after oxytocin administrations. Mares were given oxytocin, 10 or 25 iu, i.v., on the day of, or 2 days after, ovulation, and frequent jugular blood samples were collected for analysis of oxytocin and Prostaglandin F metabolite (PGFM) by RIA. Neither dose of oxytocin nor day of treatment affected the half-life of the exogenous oxytocin, which was determined to be 6.8 min. A significant increase in PGFM was observed within 6 min of oxytocin administration and peak values were observed within 10 min. PGFM response after oxytocin administration on the day of ovulation appeared elevated compared to the response 2 days after ovulation. PMID- 10454084 TI - New approaches to detect cortisol administration in the horse. AB - The cortisol threshold concentration of 1.0 microg/ml in horse urine adopted by the International Federation of the racing Authorities in 1994 is specific. However, an increase in the sensitivity for the detection of cortisol administration would be helpful. Previous studies have shown that 20beta dihydrocortisol concentration in urine would be a good indicator of cortisol administration. The purpose of the present work was to estimate the population parameters and the critical values of 20beta-dihydrocortisol and 20beta dihydrocortisone concentration in urine compared with that of cortisol. Using the same probability (1.1 x 10(-4)) which was used for the establishment of the official cortisol threshold, the critical values of 5000 ng/ml for 20beta dihydrocortisol and 350 ng/ml for 20beta-dihydrocortisone were obtained. Considering these 2 critical values for 20beta-dihydrocortisol and 20beta dihydrocortisone, the time during which a horse could be declared positive is significantly increased. PMID- 10454086 TI - Attenuation by phenylbutazone of the renal effects and excretion of frusemide in horses. AB - The objectives of this study were to determine the effect of phenylbutazone premedication on the pharmacokinetics and urinary excretion of frusemide in horses; and on frusemide-induced changes in urinary electrolyte excretion. Six Standardbred mares were used in a 3-way crossover design. The pharmacokinetics and renal effects of frusemide (1 mg/kg bwt i.v.) were studied with and without phenylbutazone premedication (8.8 mg/kg bwt per os 24 h before, followed by 4.4 mg/kg bwt i.v. 30 min before frusemide administration). A control (saline) treatment was also studied. Administration of frusemide without phenylbutazone led to diuresis, natriuresis, kaliuresis and chloruresis, and altered the ratio of sodium:chloride excretion from 0.4 to 1.0 in the first hour of diuresis. When frusemide and phenylbutazone were administered, sodium and chloride excretion in the first hour were significantly (P<0.05) reduced by 40 and 32%, respectively, when compared to frusemide administrationwithout phenylbutazone. The fractional clearance of sodium and chloride was also significantly reduced. Potassium excretion, potassium fractional clearance and the ratio of sodium to chloride excretion were not affected by administration of phenylbutazone. During peak diuresis, phenylbutazone did not affect the efficiency of frusemide with respect to electrolyte excretion. The plasma disposition of frusemide was not affected by phenylbutazone. However, the renal excretion of frusemide decreased by approximately 25%. We conclude that the decreased urinary excretion of frusemide by phenylbutazone led to an attenuation of frusemide-induced increases in urinary excretion of sodium and chloride. Since the efficiency of frusemide was not affected by phenylbutazone, we conclude that phenylbutazone attenuates the renal excretion of frusemide without inhibiting the intrarenal activity of frusemide in horses. PMID- 10454087 TI - Histopathological findings in equine sinonasal disorders. AB - Biopsies collected from 79 referred cases of equine sinonasal disease, including 27 horses with primary sinusitis, 10 with secondary dental sinusitis, 19 with sinus cysts, 11 with progressive ethmoid haematomata (PEH), 4 with false nostril epidermal inclusion cysts, 4 with sinonasal polyps, 3 with sinonasal mycosis and from 2 control animals were examined histologically. Observations were made on epithelial type and integrity, cellular inflammatory response, fibroplasia and presence of potential pathogens. Chronic inflammatory changes including mucosal thickening, ulceration and significant fibroplasia, were found in the sinus mucosa with most sinus disorders, similar to those found in human chronic sinusitis. Bacteria were variably present on sinusitis mucosae but their aetiological significance was unclear. The presence of apparently irreversible changes including fibroplasia in some of these sinusitis cases may explain their poor or delayed response to treatment. Sinus cysts had histological similarities to human mucocoeles. Progressive ethmoid haematomata showed recent and older haemorrhage, as did sinus cysts (and occasionally some chronic sinusitis sections), but support for a common aetiology between sinus cysts and PEH was absent. PMID- 10454088 TI - Characterisation of the type and location of fractures of the third metacarpal/metatarsal condyles in 135 horses in central Kentucky (1986-1994). AB - The objective of this retrospective study was to provide a detailed description of the characteristics of condylar fractures represented in a population of 135 horses who sustained 145 fractures. Records and radiographic studies were examined. Fifty-nine percent of the horses were male and the majority Thoroughbreds. The distribution of fractures was 37% incomplete-nondisplaced, 30% complete-nondisplaced and 32% complete-displaced. The right front was more likely to sustain a complete-displaced fracture, whereas the left front was more likely to sustain an incomplete-nondisplaced fracture. Forelimbs (81%) and lateral condyles (85%) were more likely to be involved. Contrary to previous studies, the right forelimb was slightly more often involved than the left. Fractures tended to involve the middle portion of the condyle (59%). The mean length of all fractures was 75+/-3.8 mm. Axial fractures and medial condyle fractures tended to be longer. Fifteen percent of the fractures had definitive articular comminution. Ninety-five percent of fractures with articular comminution were associated with complete fractures. When fractures entered the middle area of the condyle, 23% had articular comminution. Eight of the fractures spiralled, all involved forelimbs. Concurrent lesions included proximal phalanx chip fractures, sesamoid fractures, sesamoiditis, proximal phalanx fractures, 'splint' bone periostitis and ligamentous injuries. The complete description of the fractures in this group of horses allows us better to define the condylar fracture, compare these fractures to previous studies and establish new data for use in defining prognosis. PMID- 10454089 TI - Results of treatment of 145 fractures of the third metacarpal/metatarsal condyles in 135 horses (1986-1994). AB - The objectives of this study were to correlate condylar fracture characteristics and type of treatment with subsequent capacity for athletic ability, and to determine the characteristics of healing that affect prognosis after fracture fixation. Medical records, post operative radiographic studies and race records were examined for 135 horses sustaining 145 fractures. Sixty-five percent of horses overall started in a race post injury (SPI) in a mean time of 9.7 months with a mean of 13.7 races post injury. Having raced pre-injury did not confer an advantage to starting post injury, though nonstarters pre-injury tended to take longer to return. For horses starting pre- and post injury, 66% improved or maintained their race class level after injury, whereas 64.2% decreased their race earnings post injury. Eighty-five percent of the fractures received internal fixation, of which 70% were complete fractures. Eighty-seven percent of horses with incomplete-nondisplaced fractures treated conservatively raced post injury. The percent SPI for incomplete-nondisplaced, complete-nondisplaced and complete displaced fractures treated with internal fixation were 74%, 58%, and 60%, respectively. Males (72%) raced post injury more frequently than fillies (53%), and may represent a truer probability of SPI. Spiral fractures tended to take longer until their first start (mean 13.3 months). Fifty-two percent of horses with articular fragments were able to race post injury. Horses were more likely to start if 2-4 month radiographic healing revealed no evidence of the fracture except the presence of lag screws. Based on this series of cases, the majority of horses, with proper treatment, were able to return to racing regardless of fracture characteristic. Prognosis appeared to be affected by the severity of the injury to the joint, the presence of articular comminution and the quality of surgical repair. PMID- 10454090 TI - Ambient temperature and relative humidity influenced packed cell volume, total plasma protein and other variables in horses during an incremental submaximal field exercise test. AB - Thermoregulation may limit exercise performance under hot and humid conditions. This study compared heart rate (HR), respiratory rate (RR), rectal temperature (Tr), packed cell volume (PCV) and total plasma protein concentration (TPP) during a submaximal incremental field exercise test under high vs. low ambient temperature and relative humidity. Ten horses were tested 3 times in summer (July) and 3 times in autumn (September). Heart rate was measured continuously, the other variables at rest and immediately after 4 min at 3.5, 4.5 and 7.0 m/s, separated by 3 min rest intervals, and after 5 and 10 min recovery. Data for all variables were significantly greater during exercise and recovery in the hot vs. cool conditions, respectively: after 4 min at 7.0 m/s, HR was 135+/-1 and 123+/ 1/min (P<0.0001), Tr was 39.0+/-0.06 and 38.0+/-0.05 degrees C (P<0.0001), RR was 99+/-3 and 50+/-3/min (P<0.0001), PCV was 48.8+/-0.06 and 42.1+/-0.3% (P<0.0001) and TPP was 7.7+/-0.14 and 7.6+/-0.12 g/l (P = 0.026). These data reflect the thermal burden during submaximal exercise under hot conditions in the field. The greater relative PCV increase in the heat probably conferred a thermoregulatory advantage and reflected a greater circulating red cell volume increase rather than a decrease of plasma volume. This study illustrates how differences in environmental conditions can affect assessment of exercise responses and how these factors must be considered in monitoring progress during fitness and acclimatisation regimes in the field. PMID- 10454091 TI - Effect of parturition on levels of vitamins A and E and of beta-carotene in plasma and milk of mares. AB - The objective of this study was to investigate in mares the effect of parturition on plasma and milk levels of retinol, beta-carotene, alpha-tocopherol and cholesterol over 12 weeks around parturition. In blood plasma of horses around parturition an increase of all these components was observed. This increase was most impressive in beta-carotene (P<0.05) and less pronounced for vitamin E, vitamin A and cholesterol. The magnitude of increase around parturition corresponded well with the magnitude of accumulation in colostrum; levels of beta carotene in colostrum were 65 times higher compared to mature milk while vitamin A, vitamin E and cholesterol were only 3 to 8 times higher. Beta-carotene concentrations in colostrum were positively correlated with corresponding plasma levels (r = 0.9; P<0.001). Reasons for the increase in plasma beta-carotene around parturition may include an improved absorption of carotene and/or reduced conversion into vitamin A as well as mobilisation from tissue storages or a reduced uptake in tissues other than the mammary gland. In conclusion, the results may point to possible component- and species-specific differences involved in the transfer of fat-soluble vitamins, beta-carotene and cholesterol from blood plasma into colostrum. PMID- 10454092 TI - Matrix metalloproteinase-2 and -9 are activated in joint diseases. AB - A study was performed to identify the activation status of the gelatinase MMPs, MMP-2 and -9, in both normal and diseased equine articular tissues. In addition, the production and activation status of equine MMP-2 and -9 by equine articular cells and tissues in response to increasing IL-1beta concentrations was assessed. The study was performed to test the hypothesis that activation of MMPs is a fundamental step in the pathogenesis of joint diseases; and that this activation is mediated by the cytokine IL-1. Using purified equine MMP-2 and -9, the molecular weights of the zymogen and activated form of equine MMP-2 and -9 were identified by a combination of gelatin zymography and a gelatin degradation assay using aminophenylmercuric acetate as a chemical activator of the molecules. Normal equine articular tissues (cartilage and synovial membrane) maintained in short-term tissue culture produced MMP-2 zymogen alone, while similar tissues obtained from a variety of pathological conditions produce both zymogen and active MMP-2, as well as MMP-9 monomer and dimer. Activated MMP-9 was an inconsistent finding. Normal equine synovial fibroblasts in monolayer culture produced zymogen MMP-2 alone under basal conditions. A mild increase in active and zymogen MMP-2 levels occurred with IL-1beta treatment. Equine synovial membrane explants demonstrated a dose-dependent increase in active and zymogen MMP-2 and MMP-9 levels following IL-1beta treatment. Monolayer chondrocyte cell cultures demonstrated a dose-dependent mild increase in active and zymogen MMP-2 following IL-1beta treatment. Explant cartilage cultures demonstrated a dose dependent mild increase in zymogen MMP-2 alone following IL-1beta treatment. This study supports the hypothesis that activation of MMPs is occurring in joint disease, and that in vitro stimulation of equine articular cells and tissues causes not only an increase in MMP production, but also an increase in amount of activated enzyme released. Further research is required to investigate the role of MMP activation in joint diseases, and to investigate the potential use of therapeutic agents, which inhibit MMP activation, in the treatment and prevention of joint diseases. PMID- 10454094 TI - Deletion of the Sry region on the Y chromosome detected in a case of equine gonadal hypoplasia (XY female) with abnormal hormonal profiles. PMID- 10454095 TI - Depletion kinetics of clenbuterol hydrochloride in competition horses. PMID- 10454093 TI - The characteristics of intestinal injury peripheral to strangulating obstruction lesions in the equine small intestine. AB - Recent studies suggest that horses requiring surgical correction of strangulating intestinal obstruction may develop post operative complications as a result of ischaemia/reperfusion injury. Therefore, the mucosal and serosal margins of resected small intestine from 9 horses with small intestinal strangulating lesions were examined for evidence of ischaemia/reperfusion injury. Severe mucosal injury and marked elevations in myeloperoxidase activity were detected at ileal resection margins (n = 4), whereas the mucosa from proximal jejunal (n = 9) and distal jejunal (n = 5) resection margins was normal. However, the serosa from jejunal resection margins had evidence of haemorrhage and oedema, and the proximal jejunal serosa had significantly increased numbers of neutrophils. Histological injury in ileal stumps is indicative of the inability fully to resect the ileum in horses with distal small intestinal strangulations. One of 4 horses subjected to ileal resection was subjected to euthanasia and found to have a necrotic ileal stump. Evidence of serosal injury and neutrophil infiltration in the proximal jejunal resection margins may predispose horses to post operative adhesions. Four of 8 horses discharged from the hospital suffered from recurrent colic in the post operative period. PMID- 10454096 TI - Unilateral Leydig cell tumour resulting in acute colic and scrotal swelling in a stallion with descended testes. PMID- 10454097 TI - Infertility in two mares with XY and XXX sex chromosomes. PMID- 10454098 TI - Anomalous aortic origin of the left coronary artery in a horse. PMID- 10454099 TI - Depressed expression of adipocyte beta-adrenergic receptors is a common feature of congenital and diet-induced obesity in rodents. AB - OBJECTIVE: To determine whether the dramatic reduction in expression and functional activity of the beta3-adrenergic receptor (AR) and beta1AR subtypes originally observed in adipose tissue of the C57BL/6J Lep(ob)/Lep(ob) ('obese') mouse are general features of all models of obesity, and whether obesity-related differences in betaAR subtype expression occur between adipose depots. DESIGN: Survey of adipose tissue betaAR expression from four mouse models of congenital obesity: the 'obese' mouse (C57BL/6J Lep(ob)/Lep(ob)), the 'diabetic' mouse (C57BL/KsJ LepRdb/LepRdb), the 'tubby' mouse (C57BL/6J tub/tub) and the 'fat' mouse (C57BL/KsJ Cpe(fat)/Cpe(fat)), and in a model of high-fat diet-induced obesity in C57BL/6J mice. MEASUREMENTS: Expression of the betaAR subtypes was measured by Northern blot hybridization in white and brown adipose depots. RESULTS: In the severely obese Lep(ob) and LepRdb mice, mRNA concentrations of beta3AR and beta1AR in white adipose tissue (WAT) were decreased by > 99% and by > 70%, respectively. More modest effects on beta3AR expression were observed in brown adipose tissue (BAT, decreased by 20 - 30%). In less severe forms of obesity, as found in the tubby and carboxypeptidase (Cpe)fat mice, and in diet induced obese B6 mice, beta3AR expression was decreased in WAT by up to 90%, with more modest decreases in interscapular BAT (IBAT). Changes in beta1AR mRNA concentrations were more variable. Beta2AR mRNA levels did not differ in most cases, with the exception that there was a 3-5-fold increase in BAT for both Lep(ob) and LepRdb mice. CONCLUSIONS: Impaired expression of adipocyte betaAR subtypes is a general feature of both genetic and dietary obesity in mice. The degree of obesity is correlated with the extent of loss of beta3AR and beta1AR expression in WAT. The distinct endocrine abnormalities associated with these obesity models may be responsible for the degree of impaired adipocyte betaAR expression. PMID- 10454100 TI - Serum leptin concentration, body composition, and gonadal hormones during puberty. AB - BACKGROUND: Recent evidence has suggested that leptin concentration is associated with gonadal hormone levels, and that changes in leptin concentration may trigger the onset of reproductive function in children. However, the concurrent changes in body composition during puberty make the independent associations between leptin and gonadal hormone concentrations in children difficult to resolve. METHODS: To investigate the nature of associations between leptin levels and pubertal maturation, serum concentrations of leptin, estradiol, and testosterone and body composition measures were examined in a sample of 152 healthy pre pubertal, pubertal, and post-pubertal children. RESULTS: Leptin concentration was nearly three-fold higher in post-pubertal girls than in pre-pubertal girls, but was relatively similar in pre- and post-pubertal boys. Significant sex differences in leptin concentration existed in prepubertal, pubertal and post pubertal children, and these remained significant after controlling for adiposity. After adjusting for total body fat, fat-free mass and age, testosterone concentration was negatively associated with leptin levels in pubertal boys, while estradiol concentration was positively associated with leptin level in pubertal girls. CONCLUSIONS: Girls have higher serum leptin concentration before, during, and after puberty than boys, even after accounting for the development of greater female adiposity. Although other factors may be involved, sexual dimorphism in leptin concentrations during puberty appears to be partly due to a stimulatory effect of estradiol on leptin concentration in females and a suppressive effect of testosterone on leptin concentration in males. PMID- 10454101 TI - Measurement of abdominal fat by magnetic resonance imaging, dual-energy X-ray absorptiometry and anthropometry in non-obese men and women. AB - OBJECTIVE: To investigate whether the dual-energy X-ray absorptiometry (DEXA) was a better predictor of abdominal fat, measured by magnetic resonance imaging (MRI) at the level of L2-L3, than anthropometric measurements in non-obese men and non obese women. DESIGN: Observational, cross sectional study. SUBJECTS: 34 healthy subjects (17 men and 17 women) aged 20-53 y with a body mass index (BMI) < 30 kg/m2. MEASUREMENTS: Fat distribution parameters including waist circumference and waist-to-hip ratio (WHR); intra abdominal fat (IAF) by MRI; and central abdominal fat (CAF) by DEXA. RESULTS: Measurement of IAF by MRI, was highly correlated to the CAF measured by DEXA. In men, both waist circumference and WHR had similar correlation to IAF as DEXA. In women, waist circumference was less well correlated with IAF than DEXA, and the WHR had a weaker non significant correlation with IAF. CONCLUSIONS: In non-obese men; DEXA, waist circumference and WHR can predict IAF equally well, while in non-obese women, DEXA is superior to waist circumference and much better than WHR. PMID- 10454103 TI - Long term results of intensive treatment of obesity in the health centre Litija, Slovenia: a clinical audit. AB - OBJECTIVES: To explore long term results of my comprehensive group therapeutic weight-loss programme for obese people which included behavioural, psychological, cognitive and physical elements. DESIGN: Retrospective analysis of patients' records and final follow-up questionnaires filled in at least 5 y after they left their group. SUBJECTS: 48 overweight people who joined the programme were followed up for at least 5 y. 14 'non completers' did not complete the programme (median initial BMI was 32.2). Other 34 were members of groups at least 4 months (median initial BMI was 31.8). RESULTS: Median weight loss of 'completers' when they left the group was 11.5 kg. 10 members achieved normal body weight. At least 5 y later 13 of them still maintained the reduced weight. In the questionnaires they indicated that the diet, weekly group meetings, exercise and group discussions made most important contribution to their success. CONCLUSIONS: The long-term intensive and comprehensive weight loss treatment offers a good opportunity to obese people to reduce their weight and maintain it afterwards. For success they have to have adequate diet, attend the group regularly, be physically active, have support from their families and friends and change their life style and eating habits. PMID- 10454102 TI - Development of the obesity epidemic in Denmark: cohort, time and age effects among boys born 1930-1975. AB - OBJECTIVE: A global epidemic of obesity is developing, but its causes are still unclear. In Denmark, two periods of steep increases in prevalence of obesity have occurred among young men born in the 1940s and 1960-70s. This study investigated the preceding changes in prevalence of obesity and in the entire body mass index (BMI = weight/height2) distribution by birth cohort, calendar time and age among Danish school boys. METHODS: Children attending Copenhagen schools 1937-1983 had annual health examinations, from which we computerized 1,037,468 measurements of height (m) and weight (kg) of 161,314 boys aged 7-13 y. Obesity was defined as age-specific BMI exceeding the 95.0, the 99.0 and the 99.9 percentile among those born 1930-1934, the latter corresponding to the prevalence of obesity among the young men in these cohorts. The median, standard deviation, skewness, and the 5th, 25th, 75th and 95th percentiles of the age-specific BMI were estimated for each birth cohort. RESULTS: The prevalence of obesity, defined by the 99.9 percentile, increased at all ages during the same birth years as among the young men, and, accordingly, at earlier calendar years. The prevalence of obesity, defined by the 95.0 percentile, showed a distinctly different pattern: a sharp increase, irrespective of age, during the calendar years 1947-1949, and thereafter a stable level until the 1970s, where a further modest increase began. The prevalence defined by the 99.0 percentile showed a mixture of the trends in those defined by the 99.9 and 95.0 percentiles. The median BMI showed small fluctuations, parallel at all ages. The standard deviation and right-sided skewness increased until birth year 1950, but were almost stable thereafter. The pattern of changes in the quartiles mostly reflected those in the median. CONCLUSIONS: The prevalence of obesity defined by the 99.9 or 99.0 percentile has increased in Danish boys born in the 1940s and since the mid 1960s, without corresponding changes in the central part of the BMI distribution. When defining obesity by the 95.0 percentile, there was a sharp distinct age-independent increase in the late 1940s. The development of the obesity epidemic is a heterogeneous phenomenon that has involved changes in environmental influences starting at preschool ages and affecting different subsets of the population, either because of selective exposure or particular susceptibility. PMID- 10454104 TI - Time-course effects of endurance training on fat oxidation in sedentary elderly people. AB - OBJECTIVE: To investigate alterations in whole body fat oxidation after 7 and 14 weeks of progressive endurance training in sedentary elderly subjects. DESIGN: Longitudinal, 14 weeks of progressive endurance training on a cycle ergometer (3 training sessions per week). Full sets of measurements were performed before, and after 7 and 14 weeks of training. SUBJECTS: 13 healthy sedentary subjects (5 men, 8 women) (age 62.8 +/- 2.3 y). MEASUREMENTS: 24 h indirect calorimetric measurements under standardised conditions: light-activity programme, fixed food composition, neutral daily energy balance. Body composition (by isotope dilution and skinfold thicknesses). Maximal oxygen consumption. RESULTS: Loss of 0.7 kg fat mass in the first 7 weeks of training and a further 2.4 kg of fat in the second 7 weeks. There was a transient increase in sleeping fat oxidation after 7 weeks of training (+26.1%), associated with transient increase in daily fat oxidation (+/- 11.9%), but fat oxidation then returned to baseline values in the second 7 weeks. There was a correlation between within-subject changes in sleeping fat oxidation after 7 weeks of training and variations in FFM (r = 0.62, P = 0.02) and maximal oxygen consumption (r = -0.56, P < 0.05). CONCLUSION: In sedentary elderly subjects, progressive endurance training was associated with a transient increase in sleeping fat oxidation and daily fat oxidation. In free living conditions, possible changes in daily fat oxidation may have induced a negative fat balance, as judged by fat mass loss. PMID- 10454105 TI - Determinants of energy expenditure and fuel utilization in man: effects of body composition, age, sex, ethnicity and glucose tolerance in 916 subjects. AB - BACKGROUND: 24-h energy expenditure (24-EE) and 24-h respiratory quotient (24-RQ) are important measurements in obesity research, but their accurate assessment is limited to few specialized laboratories. OBJECTIVES: 1) To provide comprehensive prediction equations for 24-EE, sleeping metabolic rate (SMR) and 24-RQ, based on a large number of Caucasian and Pima Indian subjects, covering a wide range of body weight and composition, body fat distribution, and age and 2) to test whether Pima Indians have lower metabolic rate and/or higher 24-RQ than Caucasians. SUBJECTS AND METHODS: 916 non-diabetic subjects, aged 31.5 +/- 11.9 y, body weight 90.5 +/- 26.1 kg (mean +/- s.d.), (561 males, 355 females; 416 Caucasians, 500 Pima Indians; 720 with normal (NGT) and 196 with impaired (IGT) glucose tolerance) spent 24 h in a respiratory chamber for measurements of 24-EE, SMR and 24-RQ. Fat-free mass (FFM) and fat mass (FM) were assessed by either hydrodensitometry or DEXA. Waist circumference and waist-to-thigh ratio (WTR) were determined as measures of body fat distribution. RESULTS: In a stepwise multiple regression analysis, FFM, FM, sex, age, WTR, and ethnicity were significant independent determinants of 24-EE (2258 +/- 422 kcal/d), explaining 85% of its variability (24-EE (kcal/d)=696 + 18.9 FFM (kg) + 10.O FM (kg) + 180 male -1.9 age (y) + 7.1 WTR (per decimal) + 44 Pima Indian). SMR (1623 +/- 315kcal/d) was determined (78% of variability) by FFM, FM, sex, age, WTR, and glucose tolerance (SMR (kcal/d) = 443 +/- 14.6 FFM (kg) + 6.9 FM (kg) + 79 male - 1.0 age (y) + 5.8 WTR (per decimal) + 38 IGT), but not by ethnicity. Adjustment for the respective variables reduced the variance in 24-EE from 422 to 162 kcal/d and in SMR from 315 to 146kcal/d. 24-RQ (0.854 +/- 0.026) was determined by waist circumference and energy balance (24-RQ = 0.88429-0.00175 waist circumference (cm) + 0.00004 energy balance (%)), but not by sex, ethnicity or glucose tolerance. With this equation only 13% of the variability in 24-RQ could be explained (residual variance 0.024). Compared to Caucasians, Pima Indians had higher 24-EE, but similar SMR and 24-RQ. CONCLUSIONS: This analysis provides comprehensive prediction equations for 24-EE, SMR and 24-RQ from their major known determinants. It confirms the previous findings that, even after adjustment for body composition, age, sex, ethnicity, and glucose tolerance, there is still considerable variability in energy expenditure and substrate oxidation that may, in part, be genetically determined. In adult Pima Indians, we found no evidence for lower metabolic rate or impaired fat oxidation that could explain the propensity towards obesity in this ethnic group. PMID- 10454106 TI - Synergistic interactions between fenfluramine and phentermine. AB - 'Fen-phen' refers to the off-label combination of the appetite suppressants fenfluramine and phentermine. The rationale for the fen-phen combination was that the two drugs exerted independent actions on brain satiety mechanisms so that it was possible to use lower doses of each drug and yet retain a common action on suppressing appetite while minimizing adverse drug effects. The focus of the present review is to consider whether fenfluramine and phentermine exert actions that are additive in nature or whether these two drugs exhibit drug-drug synergism. The fen-phen combination results in synergism for the suppression of appetite and body weight, the reduction of brain serotonin levels, pulmonary vasoconstriction and valve disease. Fen-phen synergism may reflect changes in the pharmacokinetics of drug distribution, common actions on membrane ion currents, or interactions between neuronal release and reuptake mechanisms with MAO mediated transmitter degradation. The synergism between fenfluramine and phentermine highlights the need to more completely understand the pharmacology and neurochemistry of appetite suppressants prior to use in combination pharmacotherapy for the treatment of obesity. PMID- 10454107 TI - Prevalence of overweight and central adiposity is associated with percentage of indigenous ancestry among native Hawaiians. AB - OBJECTIVE: To assess the relationship of percentage of indigenous ancestry with body mass index (BMI) and waist to hip ratio (WHR) among Native Hawaiians. DESIGN: A cross-sectional, observational study among 567 rural native Hawaiian men and women, aged 30 or older, enrolled between 1993-96. MEASUREMENTS: Ancestry was assessed by self-report, and was verified with a brief genealogical interview. Anthropometric measurements, as well as dietary and physical activity histories were also obtained. RESULTS: Percentage Hawaiian ancestry was significantly associated with increased body mass index (BMI), waist to hip ratio (WHR), and age. The association between percentage Hawaiian ancestry and both anthropometric measures persisted after adjusting for age, leisure time physical activity and total dietary energy intake. CONCLUSION: Obesity and central adiposity were highly prevalent among Hawaiians participating in this study. The association between BMI and WHR with percentage Hawaiian ancestry, unexplained by differences in diet and physical activity, suggests genetic factors play an important role in the high prevalence of obesity observed among Native Hawaiians. PMID- 10454108 TI - Lay definitions of ideal weight and overweight. AB - OBJECTIVE: To describe lay definitions of ideal weight and overweight, to determine whether they correspond with current health definitions, and to examine the relationship between lay definitions and weight-control behaviour. DESIGN: Cross-sectional survey of a representative sample of adults from Victoria, Australia. SUBJECTS: 1342 men and women, aged 18 y and older from across the weight spectrum. MEASURES: Questionnaire-based measures of current height and weight to determine body mass index (BMI); weight that is considered to be ideal; weight that is considered to be overweight; current weight-control behaviour. RESULTS: The BMI at which women considered themselves to be at their ideal weight was significantly lower than that for men. For both men and women, the BMI defined as ideal increased with age and with current weight. The average BMI at which women considered themselves to be overweight was significantly lower than that for men, and was well within the acceptable BMI range. Just over two-thirds of men, defined overweight at a level higher than the current cutoff of 25 kg/m2. For both men and women, the BMI defined as overweight increased with age and with current weight. CONCLUSION: Lay definitions of ideal weight and overweight deviate substantially from health definitions. Public health initiatives should stress that many women are already a healthy weight, and encourage these women to focus their efforts on weight maintenance rather than weight loss. Since so few men have weight goals that are consistent with current health recommendations, it will be important to raise their awareness of what constitutes a healthy weight. PMID- 10454109 TI - Environmental factors in the development of obesity in identical twins. AB - OBJECTIVE: To study environmental factors promoting obesity when genetic factors are identical. DESIGN: Monozygotic (MZ) twins discordant for overweight were examined during a 3-day stay in an inpatient setting. SUBJECTS: The subjects were selected from the Finnish Twin Cohort. The study sample consisted of 23 healthy adult MZ twin pairs (14 female, nine male) with a difference of at least 3 kg/m2 in BMI. The mean BMI was 29.5 kg/m2 for the overweight twins and 22.9 kg/m2 for their lean co-twins. MEASUREMENTS: Interviews and standardized questionnaires were used to obtain information about energy and nutrient intake, eating behaviour, physical activity history, smoking and other background factors. RESULTS: The overweight co-twins had higher disinhibition scores (P = 0.007) and hunger scores (P = 0.005) in the 3-Factor Eating Questionnaire than their lean co twins. Among women the mean daily energy intake was higher in the overweight twins than in their lean co-twins (8.8 vs 7.4 MJ; P = 0.045). In the twins discordant for smoking the mean BMI was higher in non-smokers than in smokers (BMI 29.7 +/- 4.4kg/m2 vs 23.8 +/- 3.1 kg/m2; P = 0.031). CONCLUSIONS: Difficulty in controlling eating in both sexes and high energy intake in women were related to overweight, independent of genetic background. Smoking explained the BMI difference among smoking discordant pairs. It is probable that individual twin pairs had different reasons behind the variation in weight gain resulting in non significant intrapair differences in single obesity-promoting factors. Difference in living conditions (e.g. family-and work-related factors) may have promoted different living habits, especially eating behaviour, and may have led to different weight gain in identical twins. PMID- 10454110 TI - Gender differences in relation to leptin concentration and insulin sensitivity in nondiabetic Chinese subjects. AB - OBJECTIVE: To investigate the relationship between fasting plasma leptin concentrations and insulin resistance in Chinese men and women. DESIGN: Cross sectional study design. SUBJECTS: Ninety-six nondiabetic Chinese (51 men and 45 women) with body mass index (BMI) between 18.4-35.8 kg/m2 were studied. MEASUREMENTS: Plasma glucose and insulin concentrations were measured every 30 min for 2 h after a 75 g oral glucose load. The degree of insulin resistance was assessed using a modified insulin suppression test. Plasma leptin values were determined by radioimmunoassay. RESULTS: Fasting plasma glucose, glucose areas, fasting insulin, insulin areas, most of the lipoprotein concentrations and steady state plasma glucose (SSPG) concentrations were relatively similar between men and women. Despite the fact that men had higher BMI values (26.1 +/- 0.5 vs 24.7 +/- 0.5 kg/m2, P < 0.05), fasting plasma leptin concentrations were significantly lower in men than in women (4.9 +/- 0.5 vs 9.0 +/- 0.8 ng/ml, P < 0.001). Fasting leptin values were positively related to SSPG concentrations by simple correlation analysis in both sexes. However, this relationship persisted in men (r = 0.513, P < 0.01) but not in women (r = 0.119, P = NS) after adjustment for BMI. Multiple regression analysis showed that SSPG concentrations, BMI, glucose and insulin responses together accounted for 62.5% and 52.2% of the variation in plasma leptin concentrations in Chinese men and women respectively. CONCLUSION: Fasting plasma leptin concentrations were lower in Chinese men than in Chinese women despite the higher BMI observed in men. After adjustment for BMI, plasma leptin values correlated with the degree of insulin resistance in men but not in women. PMID- 10454111 TI - No evidence for an involvement of alleles of polymorphisms in the serotonin1Dbeta and 7 receptor genes in obesity, underweight or anorexia nervosa. AB - The serotonergic (5-hydroxytryptamine, 5-HT) system has been implicated in body weight regulation and in the etiology of anorexia nervosa (AN). Here we describe the screening of the known Phe-124-Cys polymorphism in the 5-HT1Dbeta receptor gene and of the known Pro-279-Leu polymorphism in the 5-HT7 receptor gene. For association tests allele frequencies were compared between up to 393 extremely obese children and adolescents, 142 underweight students and 84 patients with AN. None of the association tests revealed nominal P-values below 0.3. We conclude that a major role of the investigated polymorphisms in body weight regulation or AN appears unlikely. PMID- 10454112 TI - Differences in whole body measurements by DXA-scanning using two Lunar DPX-L machines. AB - OBJECTIVE: To compare bone mineral and body composition results of two dual energy X-ray absorptiometry (DXA) instruments from the same manufacturer. SETTINGS: The Medical departments of Sahlgrenska University Hospital, Goteborg and University Hospital, Uppsala, Sweden. DESIGN: Unique aluminium spine phantoms (Phant A and B) delivered with each DXA machine (DXA-A and DXA-B) were cross checked on the other machine by using the 'AP spine mode'. Eight adolescents were examined on both instruments within 5 h to ascertain total body variables by using standard, as well as extended, modes of analysis. All these double examinations were undertaken on two occasions, before and after exchange of a detector on the DXA-B. SUBJECTS: Four males and four females aged 15.4-19.2 y with normal body weights, were examined on both occasions. On each occasion, the first examination was performed in Goteborg on four individuals and in Uppsala on four individuals. RESULTS: On the first occasion the phantom measurements resulted in much lower bone mineral density (BMD) values on the DXA-B than on the DXA-A. Later it was detected that a so-called R-value and the corresponding '% fat' value were out of range on the DXA-B. After exchange of detector, the difference in phantom BMD-values between the two machines had diminished. On the first occasion in vivo BMD values were lowest on the DXA-B (P < 0.01), while on the second occasion they were significantly lower on the DXA-A (P < 0.05). Soft tissue differences were greater after detector exchange and as compared to DXA-A, DXA-B underestimated body fat by 3.5 kg (13.2 vs 9.7 kg, P < 0.001) and overestimated lean tissue mass by 3.8 kg (47.1 vs 50.9 kg, P < 0.001) on the second occasion. CONCLUSIONS: The differences in results between two apparently identical Lunar DPX-L machines were not acceptable. In multicenter studies, it may be necessary to standardise results of participating machines into results of one machine by means of regression equations obtained by examining subsamples of individuals on one master machine and other participating instruments. PMID- 10454113 TI - Body mass index (weight/height2) or percentage body fat by bioelectrical impedance analysis: which variable better reflects serum lipid profile? AB - We present population data on percentage body fat (%BF) by bioelectrical impedance analysis (BIA) in 12,287 men and 6657 women aged 30-69 y. In addition, we examined relationships among BMI, %BF and serum lipids (total-cholesterol (TC), HDL-cholesterol (HDLC), TC/HDLC ratio, LDL-cholesterol (LDLC) and triglycerides (TG)) in order to determine whether body mass index (BMI, kg/m2) or %BF by BIA better reflected lipid metabolism. Women had larger %BF than men in all age groups. On the other hand, women aged < 60 y had smaller BMI than corresponding men. Regardless of age, BMI was obviously correlated with %BF (r = 0.743-0.924). As previously reported, high BMI and high %BF were strongly associated with high serum TC/HDLC ratio and TG, and low serum HDLC rather than high serum TC and LDLC. Compared with BMI, %BF by BIA was better correlated with the serum indices except for serum HDLC. These results were found in both sexes, and there was a trend in which younger subjects had stronger correlations among BMI, %BF and serum lipids. Consequently, %BF by BIA (an index of body composition) better reflects serum lipid profile than BMI. PMID- 10454114 TI - The significance and mechanism of mitochondrial proton conductance. AB - There is a futile cycle of pump and leak of protons across the mitochondrial inner membrane. The contribution of the proton cycle to standard metabolic rate is significant, particularly in skeletal muscle, and it accounts for 20% or more of the resting respiration of a rat. The mechanism of the proton leak is uncertain: basal proton conductance is not a simple biophysical leak across the unmodified phospholipid bilayer. Equally, the evidence that it is catalysed by homologues of the brown adipose uncoupling protein, UCP1, is weak. The yeast genome contains no clear UCP homologue but yeast mitochondria have normal basal proton conductance. UCP1 catalyses a regulated inducible proton conductance in brown adipose tissue and the possibility remains open that UCP2 and UCP3 have a similar role in other tissues, although this has yet to be demonstrated. PMID- 10454115 TI - Indirect measurement of mitochondrial proton leak and its application. AB - In mitochondria, ATP synthesis is coupled to oxygen consumption by the proton electrochemical gradient established across the mitochondrial inner membrane in a process termed oxidative phosphorylation. It has long been known from stoichiometric studies that ATP synthesis is not perfectly coupled to oxygen consumption. The major inefficiency in the system is leakage of protons across the mitochondrial inner membrane driven by the proton electrochemical gradient. The kinetics of the proton leak can be determined indirectly, by measuring the oxygen consumption of mitochondria under non-phosphorylating conditions (plus oligomycin) as a function of the proton electrochemical gradient. This experimental system provides a convenient means to investigate inner membrane permeability to protons and the effect of factors that may effect that permeability. In this paper we review some results from our laboratory of indirect measurement of mitochondrial proton leak and how it has been applied to investigate the effect of aging, obesity and thyroid status on proton leak. The results show that (i) proton leak in isolated liver mitochondria is not significantly different in a comparison of young and old rats, in contrast (ii) there is an apparent increase in proton leak in in situ mitochondria in hepatocytes from old rats when compared to those from young rats, (iii) proton leak in neuronal mitochondria in situ in synaptosomes is not significantly different in young and old rats, (iv) proton leak is greater in isolated liver mitochondria from ob/ob mice compared to lean controls, (v) acute leptin (OB protein) administration restores the increased leak rate in isolated liver mitochondria from ob/ob mice to that of lean controls, (vi) administration of thyroid hormone (T3) increases proton leak in rat muscle mitochondria, and (vii) proton leak in muscle mitochondria is insensitive to the presence of GDP. It is proposed that the experimental system described here for measuring proton leak, is an ideal functional assay for determining whether the novel uncoupling proteins increase inner membrane permeability to protons. PMID- 10454116 TI - UCP1, UCP2 and UCP3: are they true uncouplers of respiration? AB - The thermogenesis in brown adipose tissue (BAT) is due to the activity of a mitochondrial uncoupling protein (UCP1). This protein allows the protons pumped by the respiratory chain to re-enter the matrix without ATP synthesis. Therefore respiration is dramatically increased and produces only heat. The discovery of genes showing strong similarities with the UCP1 gene and expressed in other tissues raised the possibility that these proteins participate in the proton leak observed in mitochondria, and therefore participate in the regulation of energy expenditure. The recombinant expression of UCP1, UCP2 and UCP3 in yeast allows the comparison of the coupling state of yeast mitochondria in the presence or absence of these proteins. PMID- 10454117 TI - Structure-function relationship in UCP1. AB - The function of uncoupling protein (UCP1) as a H+ transporter regulated by nucleotide binding is elucidated. H+ transport requires fatty acids (FA) with relatively wide structural tolerance. The nucleotide binding site is specific for purine nucleotides and tolerates a number of derivatives. The strong pH dependency facilitates regulation of nucleotide binding and thus H+ translocation. The structure-function relationship of UCP1 has been analysed by various probes and by mutagenesis. According to our model, FA are a cofactor in H+ transport, providing H+ shuttling carboxyl groups in the translocation channel. By mutagenesis, additional H+ translocating groups at both sides of the translocation channel were found. Two pH sensors, controlling nucleotide binding, were identified in accordance with earlier postulates deduced from the pH dependence of nucleoside diphosphate (NDP) and nucleoside triphosphate (NTP). A common pH sensor E190 and a specific pH sensor H214 for triphosphates only, control access to the phosphate binding moiety. The three mitochondrial carrier family characteristic intrahelical arginines are essential for nucleotide binding. Mutagenesis of other charged residues reveals their role in structure stabilisation and/or has more generalised effects due to charge relay networks in UCP1. PMID- 10454118 TI - Biochemical aspects of the uncoupling proteins: view from the chair. AB - The discovery of nonshivering thermogenesis (NST) and its location in brown adipose tissue (BAT) in the 1950s to 1970s was soon followed by purification of the first uncoupling protein (UCP1) and later by cloning of the gene for UCP1 in 1985. The properties of UCP1 fully explained the long-known phenomenon of stimulated NST in BAT. An additional four 'uncoupling proteins' have been cloned in the last two years and are in search of phenomena they can explain. The four speakers in this first session of the symposium on uncoupling proteins reviewed biochemical properties of UCP1 and of three of the novel UCPs. Several suggested functions include mediation of the mitochondrial proton leak in tissues other than BAT, therefore a major role in energy expenditure, and protection against reactive oxygen species. Tools, techniques and information not yet available and for which further research is needed are reviewed. PMID- 10454119 TI - Mitochondria uncoupling proteins and obesity: molecular and genetic aspects of UCP1. AB - Genetic variation in brwon fat specific mitochondrial uncoupling protein-1 (UCP1) expression and brown adipocyte morphology, have provided models to test the hypothesis that nonshivering thermogenesis is associated with the regulation of body weight. Genetic manipulation using transgenic animals and gene targeting, has resulted in mice with an over-expression of UCP1. These variant animals consistently show that over-expression of UCP1 reduced adiposity. On the other hand, less agreement is found in models that reduce nonshivering thermogenesis. Inactivation of the UCP1 gene, by gene targeting, does not increase adiposity when compared to control animals; however, a mouse expressing the UCP1-DTA transgene (UCPI-diphtheria toxin A chain), in which there is a modest reduction in the number of brown adipocytes, becomes obese. Other phenotypes of this mouse, the hyperphagia, extreme resistance to leptin administration, retinopathy and high residual content of brown adipocytes, suggest that the effects of the transgene may be more extensive than simply a 60% reduction in the number of brown adipocytes. Ectopic expression of UCP1-DTA in the brain could explain the phenotype of this mouse in a manner more consistent with the results of other models with altered UCP1 and brown adipocyte expression. PMID- 10454120 TI - Uncoupling protein-2 (UCP2): molecular and genetic studies. AB - Thermogenesis is associated to oxygen consumption and cellular respiration. This process is coupled to adenosine-diphosphate (ADP) phosphorylation through the existence of a proton gradient across the inner mitochondrial membrane. It was postulated that proton leaks through this membrane would uncouple respiration from adenosine-triphosphate (ATP) synthesis and induce energy dissipation as heat. Such a mechanism was identified in thermogenic brown adipose tissue mitochondria which contain a unique proton carrier referred to as uncoupling protein (UCP). This UCP is activated by fatty acids and its synthesis is positively controlled by retinoids, thyroid hormones, catecholamines and rexinoids. In fact, in most types of cells, respiring mitochondria release heat and the coupling of substrate oxidation to ADP phosphorylation is under 100%. It suggested that the partial coupling of respiration to ADP phosphorylation was due to proton leaks possibly related to the brown fat UCP. This approach led to the identification of UCP2 and UCP3, two homologues of the brown fat UCP (renamed UCP1). UCP2 gene is widely expressed in tissues and cell types, whereas the UCP3 gene is dominantly expressed in skeletal muscles (and brown fat in mice). Recent genetic, biochemical and physiological studies suggest that these novel UCP2 contribute to resting metabolic rate, fat oxidation and may represent new targets for anti-obesity compounds. PMID- 10454121 TI - Uncoupling protein-3 (UCP3): a mitochondrial carrier in search of a function. AB - UCP3 is a mitochondrial protein with high homology to the established uncoupling protein, UCP1. Its high degree of homology to UCP1 suggests that UCP3 may be a true uncoupling protein. Preliminary biochemical studies are consistent with UCP3 having uncoupling activity. However, detailed functional studies are required to understand the true biochemical and physiological purpose of UCP3. These efforts should be aided by identification of humans with inactivating mutations and/or the generation of gene knockout mice lacking UCP3. PMID- 10454122 TI - Genetics of uncoupling proteins in humans. AB - Genetic studies in humans provide a method to test hypotheses about the biological roles of specific genes. So far, ten published papers have chosen to examine the hypothesis that uncoupling protein-2 (UCP2) and/or UCP3 influence energy expenditure and/or body fat accumulation. These genes were chosen because they are candidate energy expenditure genes, based on their homology to UCP1. Studies of UCP2 and UCP3 are intrinsically intertwined because the two genes are separated by only 6000 base pairs on human chromosome 11. Linkage studies in families have suggested that UCP2 and/or UCP3, or a closely linked gene, may influence resting metabolic rate (RMR) Some association studies using a 3' untranslated region insertion/deletion variant of UCP2 have produced statistically positive evidence for association with body mass index (BMI) and RMR. In contrast, association studies of UCP2 using an Ala to Val variant at amino acid 55 have produced negative results. Positive results have also been reported for association of a UCP3 splice variant with respiratory quotient in African Americans. In addition, no studies have reported linkage or association of UCP2 or UCP3 with diabetes. Overall, the results suggest that some variants of UCP2 and UCP3 may be associated with obesity traits in some populations. The UCPs, to date, show positive results in associations with obesity traits but not with diabetes traits. Further work will be needed to settle the role of UCP2 and UCP3 alleles in human body weight regulation. PMID- 10454123 TI - The long isoform uncoupling protein-3 (UCP3L) in human energy homeostasis. AB - The biological role(s) proposed for UCP3 in energy homeostasis have been based primarily upon amino acid sequence homology to UCP1. Spontaneous mutations of UCP3> have been described in humans, but not in rodents. The functional consequences-or lack thereof-of these mutations in humans will be of great importance in elucidating the biology of this protein. The results of two such studies are summarized here. PMID- 10454124 TI - Molecular and genetic aspects of the UCPs: view from the chair. PMID- 10454125 TI - Brain distribution of UCP2 mRNA: in situ hybridization histochemistry studies. AB - Uncoupling protein-2 (UCP2) is expressed in large amounts in several tissues. In the mouse brain, in situ hybridization studies have revealed an abundant expression of UCP2 mRNA in the ventral septal region, the hypothalamus, the hindbrain (medulla), the ventricular regions and the cerebellum. In the hypothalamus, a very highly intense hybridization signal is apparent in the suprachiasmatic nucleus, in the medial parvicellular and magnocellular lateral parts of the paraventricular hypothalamic nucleus, and in the arcuate nucleus. In the brainstem, UCP2 is found to be strongly expressed in the dorsal motor nucleus of the vagus nerve. The expression of UCP2 mRNA is also clearly noticeable in the choroid plexuses and in the cerebellum. The expression of UCP2 mRNA in specific regions of the brain as well as its presence in neurons with a known chemical identity suggest that UCP2 mRNA is expressed in neurons. It is as yet premature to conclude about a specific function of UCP2 in the brain. The brain distribution pattern of its transcript suggests that this mitochondrial protein could be part of neuronal circuitries involved in the control of neuroendocrine functions and autonomic responses. Assuming that the UCP2 mRNA encodes a functional uncoupling protein, it can be argued that UCP2 contributes to the metabolic rate and thermoregulation of the neuronal structures to which it is associated. In addition, by elevating oxygen consumption in the brain, UCP2 could in specific regions control the production of reactive oxygen species and thereby influence the process of neural degeneration. PMID- 10454126 TI - Thyroid hormone and other regulators of uncoupling proteins. AB - The role of the thyroid gland in the regulation of metabolic rate has been known since the last century. The knowledge that thyroid hormones increase energy expenditure, in part by lowering metabolic efficiency, dates from the 1950s. Presumably thyroid hormones regulate energy expenditure and efficiency by controlling the rate of transcription of specific genes. However, the number, identity, and relative contributions of these genes are not known. The uncoupling proteins (UCPs) are obvious candidates to mediate thyroid thermogenesis. UCP1 is not a major contributor, since thyrotoxicosis decreases UCP1 expression and inactivates brown fat. Discovery of UCP3 and its regulation by T3 in muscle is an exciting observation, consistent with a role for UCP3 in thyroid thermogenesis. Since free fatty acids appear to regulate UCP3 expression and T3 stimulates lipolysis, further experiments are required to determine if T3 regulation of UCP3 expression is direct or not. PMID- 10454127 TI - Effects of dietary deprivation, obesity and exercise on UCP3 mRNA levels. AB - Uncoupling protein-3 (UCP3) is selectively expressed in skeletal muscle of rodents and humans, and in brown adipose tissue (BAT) of rodents. C2C12 myoblast transfection with UCP3 induced a decrease in mitochondrial membrane potential suggesting that UCP3 behaves as an uncoupler of oxidative phosphorylations. Cold exposure, food restriction and fasting affect UCP3 mRNA expression differently in BAT, compared to muscle. The effects induced by cold-exposure and fasting in BAT, and by fasting in muscle, might be explained by changes in intracellular free fatty acids (FFA). A single bout of exercise or endurance training, respectively, increases or decreases muscle UCP3 expression. The effects of PPARgamma agonists and leptin on BAT and muscle UCP3 mRNA expression are also discussed. Hypotheses to explain the effects of these modulations are presented. PMID- 10454128 TI - Uncoupling protein-2 (UCP2) and uncoupling protein-3 (UCP3) expression in adipose tissue and skeletal muscle in humans. AB - Uncoupling protein-2 (UCP2) and uncoupling protein-3 (UCP3) are mitochondrial proteins that may play a role in the control of energy expenditure by uncoupling respiration from ATP synthesis. The present review focuses on data obtained in humans. UCP2 is widely expressed in the body, whereas UCP3 expression is restricted to skeletal muscle. Positive correlations have been reported between UCP2 mRNA concentrations in adipose tissue, UCP3 mRNA concentrations in skeletal muscle, and components of the metabolic rate. Fasting induces an up-regulation of UCP2 and UCP3 mRNA expression. In vivo and in vitro studies suggest that fatty acids could modulate uncoupling protein gene expression. The putative relationship between obesity, energy expenditure and uncoupling protein expression, and the unexpected rise in UCP2 and UCP3 mRNA concentrations during short-term fasting, are discussed in view of the recent data obtained in rodents and cell lines. PMID- 10454129 TI - Obesity and increased contractile activity influence the protein content of UCP2 in human skeletal muscle. AB - The newly discovered uncoupling protein-2 (UCP2) has been proposed to play a critical role in thermoregulatory and substrate oxidation processes. Skeletal muscle mRNA expression and, more recently, the protein content of UCP2 were investigated in humans. These studies have shown that the content of this protein varies quite substantially and that several factors could be responsible for its variation in human skeletal muscle. The aim of this review is to determine whether obesity and low-intensity increased contractile activity contribute to variation in muscle UCP2 content. A recent study from our laboratories revealed that, in obesity, UCP2 content in skeletal muscle is over-expressed by about 1.5 fold compared to lean. Body weight loss in obese subjects did not cause any change in skeletal muscle UCP2 content. On the other hand, when increased muscular contractile activity of knee extensor muscles is induced by several weeks of low-frequency electrical stimulation, UCP2 content increased by about 15%. Obesity and increased contractile activity do not appear sufficient, however, to explain the magnitude of the human skeletal muscle variation in UCP2 content. Since intensive efforts are being devoted to this area of research, it is expected that our understanding of the causes contributing to its variation in humans will soon be substantially improved. PMID- 10454130 TI - The physiological role of the novel uncoupling proteins: view from the chair. AB - The recently cloned uncoupling proteins -2 and -3 (UCP2, UCP3) cDNAs encode for proteins with 57-59% homology with brown adipose tissue uncoupling protein (UCP1) that can function as uncouplers of phosphorylation. The presentations of this Session of the Symposium were largely based on mRNA measurements, with one study showing that protein concentrations are well predicted by changes in mRNA in human muscles. Overall the observations suggest more complexity than anticipated. These observations are for the moment hard to reconcile with a simple role for these proteins in energy dissipation. Just the marked differences in tissue distribution of the two novel UCPs suggest they have different functions. It is possible that the activity, rather than the concentrations of these proteins, is regulated. The investigation of this possibility will hopefully enlighten us on their physiological role. PMID- 10454131 TI - The Ile93Met mutation in the ubiquitin carboxy-terminal-hydrolase-L1 gene is not observed in European cases with familial Parkinson's disease. AB - Recently an Ile93Met mutation in the ubiquitin-carboxy-terminal-hydrolase-L1 gene (UCH-L1) has been described in a German family with Parkinson's disease (PD). The authors showed that this mutation is responsible for an impaired proteolytic activity of the UCH-L1 protein and may lead to an abnormal aggregation of proteins in the brain. In order to determine the importance of this or any other mutation in the coding region of the UCH-L1 gene in PD, we performed mutation analysis on Caucasian families with at least two affected sibs. We did not detect any mutations in the UCH-L1 gene, however, we cannot exclude mutations in the regulatory or intronic regions of the UCH-L1 gene since these regions were not sequenced. We conclude that the UCH-L1 gene is not a major gene responsible for familial PD. PMID- 10454132 TI - Homologous regulation of 5-HT1A receptor mRNA in adult rat hippocampal dentate gyrus. AB - Short-term adrenalectomy induces a loss of mature granular neuronal phenotypes in the hippocampal dentate gyrus; injection of 5-HT1A receptor agonist reverses this effect. Adrenalectomy also induces an increase of expression of 5-HT1A receptor mRNA in the dentate gyrus. This study tested the effect of 5-HT1A agonist on this adrenalectomy-induced increase of 5-HT1A mRNA. Five, 9 and 18 days after adrenalectomy, 5-HT1A receptor mRNA is increased in the granular layer of the dentate gyrus. The increase is nearly 100% at day 18 after adrenalectomy. 5-HT1A agonist treatment decreased 5-HT1A mRNA both at 9 (20%) and 18 days (34%) after adrenalectomy. Our results indicated that a 5-HT1A agonist can partly reverse the adrenalectomy-induced increase of 5-HT1A mRNA and loss of mature granular neuronal phenotypes in hippocampal dentate gyrus. PMID- 10454133 TI - An immunohistochemical investigation of the opioid cell column in lamina X of the male rat lumbosacral spinal cord. AB - Tri-color immunohistochemistry was employed to examine enkephalin-like immunoreactive neurons in lamina X of the rat lumbosacral spinal cord. Serial coronal sections from levels L1 to S3 were examined. A rostral group of large (40 50 microm diameter), pyramidal-shaped enkephalin-like immunoreactive neurons were shown from levels L1 to L4-5. Essentially all of these neurons were also immunoreactive for galanin and cholecystokinin. A second enkephalin-like immunoreactive cell group, extending from L5 to approximately the S2-3 level, contained smaller (20-30 microm diameter), ovoid-shaped perikaryia. Approximately 75% of these enkephalin-like immunoreactive neurons were also immunoreactive for neuropeptide Y. Neurotensin-immunoreactivity was also present in this area, having varying amounts of co-localization with these other two peptides. These results demonstrate that the lumbosacral opioid cell column in lamina X is not a neurochemically homogenous structure. PMID- 10454134 TI - Activation of human language processing brain regions after the presentation of random letter strings demonstrated with event-related functional magnetic resonance imaging. AB - This study addresses the question, to what extent the processing of meaningless random letter strings involves classical language related brain regions. Using event-related functional magnetic resonance imaging (fMRI), which allows random stimulus presentation, we could demonstrate activation in the left inferior frontal gyrus, the left superior temporal gyrus, left parietal and occipital regions after the presentation of random letter strings compared to real words. The activation in these classical language related brain areas reflects an intense lexical evaluation process of the meaningless stimuli. Real words contrasted with random letter strings activated the left angular gyrus, bilateral precuneus and the left posterior cingulate gyrus, which may reflect the access of higher semantic associations. PMID- 10454135 TI - Slow cortical potential shifts modulate the sensory threshold in human visual system. AB - The relationship between the spontaneous slow cortical potential shifts and the detection of visual stimuli at sensory threshold were investigated. The mean slow cortical potentials preceding the detected stimuli were more negative than those preceding the missed stimuli. Accordingly, the stimulus detection performance was higher during negative compared to positive cortical potential shifts. These findings demonstrate that the cortical negativity reflects increased excitability of neural networks, thereby facilitates the detection of threshold stimuli, in contrast to cortical positivity. Therefore, at near-threshold stimulus intensities one reason for detecting the stimulus in one trial but missing it in another could be the change in the EEG baseline between the trials. PMID- 10454136 TI - Differences in human visual evoked potentials during the perception of colour as revealed by a bootstrap method to compare cortical activity. A prospective study. AB - The aim of this prospective study was to investigate the colour related information in cortical activity as it is recorded from the scalp, by comparing the shape of the potential fields. Six healthy volunteers and two volunteers with known protanopsia were used to record multichannel visual evoked potentials after stimulation with chromatic stimuli of equally perceived brightness. The scalp fields resulting from each of the four chromatic stimuli were compared in pairs, in every possible combination and for each time point, using Efron's bootstrap method. It was found that, in comparison to other stimuli responses, the long dominant wavelength stimulus results in significant differences of cortical activity. These are mainly identified in two time periods: (a) at mid-latency responses, usually during the onset and development of P100 component, and (b) after the peak (on the decline) of P100 component. Similar but less evident behaviour was identified when the responses from the short dominant wavelength stimulus were compared with those from the other stimuli. Colour effects were not significant in protanops. The proposed method can be used to locate in time and quantify the differences in cortical activity during colour perception. PMID- 10454137 TI - Neutralizing antibodies to interleukin 1-receptor reduce pain associated behavior in mice with experimental neuropathy. AB - We investigated whether interleukin-1 (IL-1), a mediator of inflammatory pain, also plays a role in pain induced by nerve injury. Female C57BL/6-mice with a chronic constrictive injury of one sciatic nerve, an established model of neurogenic hyperalgesia and allodynia, were treated with different doses (10-80 microg) of a neutralizing monoclonal rat antibody to IL-1 receptor I (anti-IL 1RI). This antibody dose-dependently reduced thermal hyperalgesia and mechanical allodynia in the animals. Furthermore, immunoreactivity for the proinflammatory cytokine tumor necrosis factor-alpha (TNF) was reduced in mice treated with the highest dose of anti-IL-1RI. Degeneration of myelinated fibers was not altered by any of the treatment schedules. We conclude that IL-1 may be a mediator of hyperalgesia after nerve lesion. PMID- 10454138 TI - Reversal of thermal hyperalgesia in a rat partial sciatic nerve ligation model by Prosaptide TX14(A). AB - We used the partial sciatic nerve ligation (PSL) model of Seltzer to assess thermal hyperalgesia after administration of Prosaptide 14-mer, TX14(A). At a dose of 200 microg/kg in Wistar rats, subcutaneous delivery of TX14(A) reversed thermal hyperalgesia at 3 and 24 h. Values declined at 48 h and returned to baseline at 72 h. A dosing study of TX(14)A gave a dependent response with 100 microg/kg having a similar potency to the 200 microg/kg study with 50 and 10 microg/kg responding somewhat lower. When TX(14)A was administered every fourth day for 12 days at 100 microg/kg, 24 h post injection values returned to baseline each time. Our results suggest that Prosaptide may have potential for therapeutic use in neuropathic pain syndromes in humans. PMID- 10454139 TI - Raphe pallidus and parapyramidal neurons regulate ear pinna vascular conductance in the rabbit. AB - We have determined whether alteration of neuronal function in raphe pallidus and the parapyramidal region alters ear blood flow, measured by an implanted Doppler ultrasonic probe, in anesthetized rabbits. Injection of GABA (5 nmol in 50 nl) increased ear flow from 6.0 +/- 1.0 to 31 +/- 10 kHz, without changing arterial pressure or heart rate. Focal electrical stimulation of raphe pallidus at low current amplitude caused ear pinna blood flow to fall from 41 +/- 6 to 9 +/- 3 kHz, again with little or no change in arterial pressure. These excitatory and inhibitory stimuli did not affect superior mesenteric blood flow. The fall in ear flow in response to electrical stimulation of raphe pallidus was not prevented by tetrodotoxin-mediated inhibition of the rostral ventrolateral medulla. Thus raphe pallidus and parapyramidal region may regulate ear pinna vascular conductance via a direct spinal projection. PMID- 10454140 TI - Imaging functional activation of the auditory cortex during focal repetitive transcranial magnetic stimulation of the primary motor cortex in normal subjects. AB - Positron emission tomography (PET) during focal repetitive transcranial magnetic stimulation (rTMS) has emerged as a promising approach to study cortical connectivity in awake humans. However, the noise caused by the discharging magnetic coil might have confounding effects on the rTMS-related cortical activation pattern. In twelve healthy volunteers, 18-fluoro-2-deoxy-D-glucose (18FDG) PET was employed to visualize the functional activation of the primary auditory cortex (PAC) during 2 Hz rTMS of the left primary sensorimotor hand area. Magnetic stimuli (1800) were applied at an intensity of 140% of motor resting threshold during the uptake period of 18FDG. Though all subjects wore earplugs, rTMS-related noise induced a consistent bilateral increase of regional glucose utilization in the PAC (P < 0.05, corrected). Thus, rTMS-related acoustic input needs to be taken into account in combined rTMS/PET studies. PMID- 10454141 TI - Expression of wild-type and V210I mutant prion protein in human neuroblastoma cells. AB - The conversion of the host-encoded prion protein (PrPc) into the insoluble, protease-resistant isoform (PrPsc) is the main pathogenic mechanism of transmissible spongiform encephalopathies. They are fatal neurodegenerative disorders, which in human occur as sporadic, inherited or familial forms. These last forms are linked to insert or point mutations of PrPc which may facilitate the spontaneous conversion into PrPsc. We have established stably transfected human neuroblastoma cells (SH-SY5Y) expressing mutant V210I, or wild-type PrPc. Both proteins were expressed and attached to the cell surface. The mutation in position 210 did not alter the biochemical properties of the protein in comparison with the wild-type protein nor induced any conformational changes similar to those observed in PrPsc. PMID- 10454142 TI - Brain-derived growth factor and nerve growth factor concentrations are decreased in the substantia nigra in Parkinson's disease. AB - Using highly sensitive sandwich enzyme-linked immunosorbent assays (ELISA), we measured for the first time the concentrations of brain-derived growth factor (BDNF) in the brain (substantia nigra, caudate nucleus, putamen, cerebellum, and frontal cortex) from control and parkinsonian patients. BDNF in the human brain (the order of ng/mg protein) was significantly lower specifically in the nigrostriatal dopamine (DA) regions from parkinsonian patients than in those from control patients. The concentration of nerve growth factor (NGF) was also significantly decreased in the substantia nigra of parkinsonian patients in comparison with that in the controls. Since BDNF and NGF may play important roles in survival and differentiation of neuronal cells, the present data indicate that the lack of neurotrophins, especially BDNF, may be involved in the pathogenesis of PD during progress of neurodegeneration of the nigrostriatal DA neurons. PMID- 10454143 TI - Differential changes in the subcellular distribution of alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionate and N-methyl-D-aspartate receptors in neonate and adult rat cortex. AB - We compared the distribution of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors and their individual subunits in synaptosomal and microsomal fractions prepared from 2-day-old (P2) and adult rat cortex. In P2 cortex more [3H]-(S)-fluorowillardiine ([3H]FW) binding to AMPA receptors was in the intracellular microsomal fraction than in the synaptosomal fraction whereas in adult rats the reverse was observed. Immunoblots with GluR1, GluR2/3, GluR4 and pan-AMPA antibodies showed the same profile. In contrast, the majority of [3H]MK-801 binding to the N-methyl-D-aspartate (NMDA) receptor and NR1 subunit immunoreactivity was present in the synaptosomal fraction at both developmental time points. These results suggest a developmental rearrangement of the distribution of AMPA receptors within neurons, a process which is likely to be important in synaptic stabilization and plasticity. PMID- 10454144 TI - In situ localization of neuronal nitric oxide synthase (nNOS) mRNA in the rat retina. AB - We performed a comparative study on the distribution of neuronal nitric oxide synthase (nNOS) immunoreactivity and mRNA in a normal rat retina using immunocytochemistry and in situ hybridization technique. As in previous studies, we found NOS immunoreactive (NOS-IR) cells and fibers in inner and outer plexiform layers (IPL and OPL), inner nuclear layer (INL) and inner photoreceptor segment (IPS). However, very little nNOS-IR could be detected in groups of amacrine cells and ganglion cells localized in ganglion cell layer (GCL). However, in situ hybridization showed that intense NOS mRNA signals were mainly found in the GCL and INL while weak or no mRNA signals were detected in IPL, OPL, outer nuclear layer (ONL) and IPS. This difference suggests that nNOS proteins may be transported through axons into the terminals in the IPL and OPL after they were synthesized with nNOS mRNA templates in the INL. In the case of nNOS mRNA in GCL, synthesizing nNOS proteins may move outside the eyeball and carry out tasks in central nervous system. The difference of nNOS mRNA and nNOS IR means that the complete concurrence of nNOS IR and in situ hybridization results may not always occur in the rat retina. PMID- 10454145 TI - Serotonin depolarizes hippocampal interneurones in the rat stratum oriens by interaction with 5HT2 receptors. AB - Patch-clamp recording techniques were used to examine the effect of serotonin (5HT) upon interneurones contained in the stratum oriens layer of hippocampal slices. Bath application of 1-20 microM 5HT depolarized neurones by the induction of an inward current at -60 mV. This inward current was Na+-dependent in nature, was mimicked by the 5HT2 receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2 aminopropane (DOI) and was inhibited by pre-incubation with the 5HT2 receptor antagonist ritanserin. PMID- 10454146 TI - Lysophosphatidic acid-induced, pertussis toxin-sensitive nociception through a substance P release from peripheral nerve endings in mice. AB - The intraplantar injection of lysophosphatidic acid (LPA) at doses of 0.1-100 pmol into the hind limb of mice showed dose-dependent nociceptive flexor responses. Repeated challenges of LPA at 100 pmol every 5 min showed constant responses at least for 30 min. The prior application of pertussis toxin (PTX) at a dose of 10 ng markedly reduced the following LPA (100 pmol) actions. In addition, the intraplantar application of CP-99994 (1 pmol), a substance P (NK1) receptor antagonist, but not CP-100263 (1 pmol), an inactive derivative, also markedly reduced the LPA responses. These findings suggest that LPA has a nociception-producing activity on sensory neurons through G(i/o) activation and substance P release from nociceptor endings. PMID- 10454147 TI - Structure and alternative splicing of the gene encoding alpha1I, a human brain T calcium channel alpha1 subunit. AB - The structure of CACNA1I, the gene encoding alpha1I, a human brain T Ca2+ channel alpha1 subunit, was determined by comparison of polymerase chain reaction amplified brain cDNA and genomic sequences. The gene consists of at least 36 exons spanning at least 115,168 basepairs of chromosome 22q12.3-13.2. The predicted protein has 2016 amino acids and 28 potential phosphorylation sites. Alternative splicing of the gene occurs at two sites: cassette exon 9 and an alternative acceptor in exon 33. Molecular diversity generated by alternative splicing and post-translational modification of this and other members of the T alpha1 subunit gene family may account for the observed heterogeneity of T currents in central neurons. PMID- 10454148 TI - Chondroitin sulfate E promotes neurite outgrowth of rat embryonic day 18 hippocampal neurons. AB - In light of controversial reports concerning the effects of chondroitin sulfates on neurite outgrowth, several glycosaminoglycans belonging to this structural class were compared with regard to their influence on axon formation by embryonic day 18 hippocampal neurons. In these studies, chondroitin sulfate A (CS-A), CS-B and CS-C proved weak or inefficient in the neurite outgrowth promotion assay. As expected, CS-D stimulated both the fraction of neurite bearing neurons and the length of their processes. This effect could be neutralized by the monoclonal antibody (mAb) 473HD. In contrast, CS-E enacted a dramatic promotion of neurite outgrowth. This effect persisted in the presence of mAb 473HD, consistent with the observation that this antibody did not react with CS-E in glycosaminoglycan transfer and blotting techniques. We conclude that CSE contains a novel glycosaminoglycan based neurite outgrowth promoting motif, which is distinct from other known activities. PMID- 10454149 TI - Implicit memory within a word recognition task: an event-related potential study in human subjects. AB - First, we recorded brain potentials from 15 healthy young subjects during the performance of a word/non-word discrimination task. During continuous visual presentation, some of the meaningful words were repeated after 86-94 s. We found a significant decrease of response time associated with the classification of repeated words which is an index for priming, an unconscious brain process. However, event-related potentials (ERPs) did not differ significantly between first and second presentations. Second, we recorded brain potentials during a following recognition test. Some of the meaningful words which were presented only once during the semantic discrimination task were repeated and had to be discriminated from randomly interspersed new words. We compared ERPs produced by incorrectly classified repeated words (misses) with ERPs produced by correctly classified new words (correct rejections). We found early ERP differences between 250 and 400 ms and later differences starting at about 500 ms after the stimulus onset. The early effect occurred over parietal scalp locations and the later effect over frontal, parietal and occipital scalp locations. This is evidence for unconscious brain activity related to the processing of missed repeated words. We suggest that the later frontal effect we found is due to an enhanced effort of the retrieval of item representations during word recognition and that the earlier parietal effect reflects partial recognition. PMID- 10454150 TI - Effect of dopamine receptor anatagonists on the calcium-dependent central function that reduces blood pressure in spontaneously hypertensive rats. AB - The effects of intracerebroventricular (i.c.v.) administration of dopamine receptor antagonists on the calcium-dependent brain function that reduces blood pressure were investigated. The systolic blood pressure of spontaneously hypertensive rats (SHR; male, 13 weeks of age) was reduced following i.c.v. administration of calcium chloride (100 microg/rat), and this effect of calcium chloride was attenuated by i.c.v. injection of eticlopride (dopamine D2 receptor antagonist, 100 microg/rat), but not by i.c.v. injection of SCH 23390 (dopamine D1 receptor antagonist, 30 microg/rat). Taking into consideration these results with our previous reports, it is suggested that calcium enhances dopamine synthesis in the brain through a calmodulin-dependent system, and that the resultant increase in dopamine levels inhibits sympathetic activity via the dopamine D2 receptor in the brain and reduces the blood pressure in SHR. PMID- 10454151 TI - Differences in the ability to process a visuo-spatial task are reflected in event related slow cortical potentials of human subjects. AB - Recent positron emission tomography (PET) and electroencephalographic (EEG) studies suggest that higher ability in a cognitive task is associated with a more efficient neuronal processing of this task. However, the validity and generalizability of these studies is limited for several reasons. We investigated 20 male and 18 female human subjects with good vs. poor spatial ability performing a visuo-spatial task (cube test). Processing-related slow event related potentials were recorded via 22 electrodes, evenly distributed over the scalp. Significant differences between good and poor performers were found in both sexes: poor subjects showed higher activity in the parietal region, and their topography was more extended into fronto-central regions. Since the amount and topography of brain activity may vary considerably depending on subjects' ability, we conclude that careful (experimental) control of task-specific ability of subjects is mandatory for cognitive neuroscience studies. PMID- 10454152 TI - Effects of age and sex on the water maze performance and hippocampal cholinergic fibers in rats. AB - We have examined if age-related deterioration of spatial memory and cholinergic innervation of the dentate gyrus is gender-specific. Aging progressively affected the performance of male and female rats in place discrimination version of the water maze task. On repeated acquisition task, only old males, but not old females, were significantly impaired relative to young and adult animals of both sexes. In parallel, we found that the age-associated reduction of the density of cholinergic fibers in the dentate gyrus was significantly more profound in old males than in age-matched females. These results suggest that, although male and female rats have an identical pattern of reference memory decline, impairment of the working memory and deterioration of the hippocampal cholinergic system are slower to develop in females than in males. PMID- 10454153 TI - Increased calcium/calmodulin protein kinase activity in astrocytes chronically exposed to ethanol: influences on glutamate transport. AB - The effects of ethanol exposures on calcium/calmodulin-dependent protein kinase activity as well as its influence on glutamate uptake were determined in astrocytes prepared from neonatal rat cerebral cortex. Acute 15-min exposure to 100 mM ethanol had no effect on Ca2+/CaM-dependent protein kinase activity. However, chronic exposure to 100 mM ethanol for 4 days elicited a significant increase in the activity of this enzyme with no parallel increase in its expression. Ca2+/CaM-independent kinase activity was less than 1% of the Ca2+/CaM dependent kinase activity and was unaffected by any of the ethanol exposures. Exposure to 100 mM ethanol for four days also resulted in a significant increase in Na+-dependent [3H]glutamate uptake which was reversed when ethanol-exposed astrocytes were co-incubated with KN-93, a specific inhibitor of Ca2+/CaM kinase. These results suggest that the effects of ethanol on glutamate transport may be mediated in part, by the level of Ca2+/CaM kinase activity. PMID- 10454154 TI - Affective properties of intra-medial preoptic area injections of testosterone in male rats. AB - On alternating days, adult male Long-Evans rats implanted with unilateral cannulae in the medial preoptic area (MPOA) received intracerebral injections of testosterone (0.05, 0.1 or 0.2 microg/0.5 microl), or saline immediately prior to confinement for 30 min to one of two compartments of a place preference apparatus. All rats received 8 days of pairings (4 hormone and 4 saline). On day 9, the rats were given a hormone-free 20-min test session during which they had access to all compartments of the apparatus. Intra-MPOA injections of testosterone (0.1 microg) produced a conditioned place preference, while injections of a higher dose (0.2 microg) produced a conditioned place aversion. The rewarding effects of intra-MPOA testosterone may in part mediate the facilatory effects of testosterone on motivational aspects of sexual behavior. PMID- 10454155 TI - Visually guided motor imagery activates sensorimotor areas in humans. AB - Stimulus-related changes in ongoing electroencephalography (EEG) over sensorimotor areas were investigated during a visually cued motor imagery task. Four subjects were instructed to imagine one-sided hand movements in response to visual cue stimuli. The EEG was recorded from central areas using 27 electrodes set at distances of 2.5 cm. The method of common spatial filters was used to extract discriminatory information of EEG patterns recorded during the two motor imagery conditions. Single EEG trials were classified in intervals of 250 ms for a 8-s period starting 3 s prior to stimulus presentation. The results suggest that perception of the visual cue stimulus modifies oscillations in sensorimotor areas specific to the indicated hand starting as soon as 250-500 ms after stimulus onset. PMID- 10454156 TI - Sensitization by prolonged glutathione depletion of kainic acid to potentiate DNA binding of the nuclear transcription factor activator protein-1 in murine hippocampus. AB - In four of four mice intracerebroventricularly injected with the inhibitor of glutathione synthesis L-buthionine-[S,R]-sulfoximine (BSO) 2 days before, an intraperitoneal injection of kainic acid (KA) invariably led to marked potentiation of DNA binding activity of the nuclear transcription factor activator protein-I (AP1) in the hippocampus at a dose which was ineffective in animals previously injected with vehicle alone. However, KA failed to potentiate binding in animals injected with BSO 1 day before. The intracerebroventricular injection of BSO induced marked and prolonged depletion of a total glutathione content in murine hippocampus for 1-2 days after administration. These results suggest that prolonged depletion of endogenous glutathione for a period longer than 1 day may lead to sensitization of KA signals to potentiate AP1 DNA binding in cell nuclei and thereby modulate de novo synthesis of particular proteins at the level of gene transcription in murine hippocampus. PMID- 10454157 TI - The effects of morphine on supraspinal and propriospinal somatocardiac reflexes in anesthetized rats. AB - In anesthetized rats, the effects of i.v. injection of morphine on supraspinally- and spinally-mediated tachycardiac reflex responses of heart rate (HR) and cardiac sympathetic nerve reflex activity were examined following electrical stimulation of either a non-segmental hindlimb (tibial) or a segmental (3rd or 4th intercostal, IC3-4) afferent nerve. In central nervous system (CNS)-intact rats, the supraspinally-mediated reflex increase in HR in response to tibial afferent nerve stimulation was augmented by morphine, whereas the increase in response to IC3-4 afferent nerve stimulation was variable. Both the supraspinally mediated myelinated (A) and unmyelinated (C) reflex discharges in the cardiac sympathetic nerve elicited by tibial afferent nerve stimulation were augmented by morphine. The effects of morphine on A- and C-sympathetic reflex discharges elicited by IC3-4 afferent nerve stimulation varied depending upon whether the discharge was of supraspinal or spinal origin. In spinal rats (spinalized at the cervical level), tibial afferent nerve stimulation did not produce any HR response, whereas IC3-4 afferent nerve stimulation produced a reflex increase in both HR and sympathetic reflex discharges of spinal origin. Furthermore, these spinal HR and sympathetic nerve reflex responses were inhibited by morphine. In conclusion, morphine depressed somatocardiac sympathetic reflexes at the spinal level, but enhanced these reflexes at the supraspinal level, leading to different effects of morphine on somatically-induced HR responses of supraspinal and spinal origins depending upon the segmental levels of afferent nerves stimulated. PMID- 10454159 TI - STAT3-mediated astrocyte differentiation from mouse fetal neuroepithelial cells by mouse oncostatin M. AB - The interleukin (IL)-6 family of cytokines share gp130 as a signal-transducing receptor component. We here show that oncostatin M (OSM), a member of this family, and its receptor components were expressed in embryonic mouse brain and that OSM induced astrocytes in cultured fetal mouse neuroepithelial cells. OSM induced promoter activation of the gene for an astrocyte marker, glial fibrillary acidic protein. The activation was completely blocked by expression of a dominant negative form of STAT3, a transcription factor activated downstream of gp130, or by introduction of a mutation in the STAT3 binding motif in the promoter. Taken together with its expression, we suggest that OSM contributes to the induction of astrocyte differentiation in a fetal developing brain via STAT3 activation. PMID- 10454158 TI - Ketamine suppresses c-fos expression in dorsal horn neurons after acute constrictive sciatic nerve injury in the rat. AB - Fos immunoreactivity within the spinal cord in a model of neuropathic pain was studied. Dorsal horn neurons in laminae I and II exhibited selective expression within the tibial, peroneal and posterior cutaneous nerve territories which, in turn, was suppressed during ketamine but not halothane anesthesia. Fos immunoreactive neurons have a unique response pattern to neuropathic pain which is sensitive to ketamine. PMID- 10454160 TI - Apolipoprotein E promoter and alpha2-macroglobulin polymorphisms are not genetically associated with Chinese late onset Alzheimer's disease. AB - In this study, we investigated two newly reported polymorphisms in association with late onset Alzheimer's disease (AD) in Chinese. They were a -491 A/T polymorphism in the Apolipoprotein E (APOE) promoter region and a five base pair deletion at exon 18 of alpha2-Macroglobin (A2M). There were 196 AD and 180 normal controls (N), which were age- and sex-matched. APOE epsilon4 alleles were significantly increased in AD vs. N (chi2 = 33.3, P < 0.000001). However, neither the -491 A/T (chi2 = 1.13, P = 0.29) nor A2M (chi2 = 0.18, P = 0.67) polymorphism was associated with AD risk, suggesting that these polymorphisms do not represent risk factors for AD in the Chinese population. PMID- 10454161 TI - Glial cell line-derived neurotrophic factor protects dopaminergic neurons from 6 hydroxydopamine toxicity in vitro. AB - Glial cell line-derived neurotrophic factor (GDNF) is a potent and specific neurotrophic factor for dopaminergic neurons. GDNF has been previously shown to protect dopaminergic neurons from lesion-induced degeneration in vivo. In this study we investigated the effect of GDNF on 6-hydroxydopamine (6-OHDA)-treated dopaminergic neurons in vitro. In dissociated cultures of embryonic rat mesencephalon, 6-OHDA exhibited a dose-dependent cytotoxicity on tyrosine hydroxylase (TH)-immunoreactive neurons. After pre-treatment with GDNF, however, 6-OHDA-induced loss of dopaminergic neurons was effectively reduced. It has been shown recently that GDNF signals through the receptor tyrosine kinase Ret and the GDNF receptor-alpha (GFR-alpha). By RT-PCR, we found both Ret- and GFR-alpha genes to be expressed in the cultured mesencephalic cells. We propose that the neuroprotective effect of GDNF on 6-OHDA-treated dopaminergic neurons in vitro is most likely mediated by functional Ret receptor signaling pathways. PMID- 10454162 TI - Glycine-immunoreactive synapses in the central nucleus of the inferior colliculus. An electron microscopic study in the cat. AB - It has been known that the inferior colliculus contains many glycinergic fibers of both intrinsic and extrinsic nature. In the present study, glycine immunoreactive (Gly-ir) synapses were examined in the central nucleus of the inferior colliculus in the cat. About half of 891 axondendritic synapses that were identified in the present study were Gly-ir. In 75% of these Gly-ir synapses, synaptic axon terminals contained pleomorphic or flattened synaptic vesicles and made symmetric synapses, while in 25% of Gly-ir synapses, synaptic axon terminals were filled with spherical synaptic vesicles and formed asymmetric synapses. Thus, Gly-immunoreactivity was detected in axodendritic synapses that formed both Gray's type I and Gray's type II synapses. PMID- 10454163 TI - Patients with palpitations referred for 24-hour ECG recording. AB - Palpitations are one of the most common reasons for referral to cardiologists; most are assessed as being medically benign. Eighty consecutive subjects referred for 24-hour ECG recording completed a questionnaire at the time of assessment and a follow-up self-report questionnaire at 18 months. Many of those with and without abnormal hearts reported distress, concern about their symptoms, and limitation of everyday activities. There were few differences between these groups at assessment or at follow-up. There was little change in mental state or in disability in either group over 18 months. Present symptomatic treatment is largely ineffective and there is a need to evaluate psychological and behavioral interventions. PMID- 10454164 TI - Is an organic/functional distinction psychologically meaningful in patients with dysphonia? AB - Dysphonia (hoarseness) is a common clinical condition and, if persistent, patients are referred to otolaryngology clinics for clinical examination. During the examination, a clinical distinction is often made among three types of patients: (1) those with a clear organic basis for dysphonia (cancer, vocal cord palsy): (2) those with some degree of organic pathology; and (3) those with an apparently functional etiology. Functional patients are often characterized as having a psychogenic disorder. This study assessed the psychological validity of the functional category in 204 out-patients (aged 17 to 87 years) with persistent hoarseness of types (2) and (3). Following clinical examination, a consultant otolaryngologist categorized patients as having functional or organic etiology. Subjects were then compared on measures of personality and psychological distress. Dysphonic subjects showed marked psychological distress compared with norms, and reported significantly more previous psychosomatic symptoms than norms, but there were no differences in personality or psychological distress between organic and functional subgroups of dysphonics. PMID- 10454165 TI - Toward empirically based criteria for the classification of somatoform disorders. AB - There is a major need for an empirical evaluation of classification criteria for somatoform disorders. The present study analyzes psychometric properties of the existing criteria for somatization disorder. The full sample consisted of 324 patients seeking help because of "psychosomatic problems." Data from a subsample of carefully diagnosed patients with somatization syndrome (n = 76) and a clinical comparison group (n = 32) permitted the analysis of the discriminative power of items. Twenty-one somatic symptoms adopted from DSM-IV and ICD-10 criteria did not exhibit the necessary psychometric characteristics (item probability, item-total correlation, etc.). Thirty-two somatic symptoms showed a satisfactory psychometric performance. A cut-off of seven or more symptoms yielded the best discrimination between low and high disability. New criteria for somatization syndrome ("polysymptomatic somatoform disorder") are proposed taking into account for the strong association of somatization and abnormal illness behavior. PMID- 10454166 TI - Culturally informed psychosomatic research. PMID- 10454167 TI - Understanding taijin kyofusho through its treatment, Morita therapy. AB - This article presents a brief review of the definition, nosology, history, clinical features, and etiology of taijin kyofusho. Special attention is also given to Morita therapy for taijin kyofusho. The term taijin kyofusho literally means the disorder (sho) of fear (kyofu) of interpersonal relations (taijin). Morita therapy was developed by Masatake Morita in the 1910s to treat the Japanese mental disorders called shinkeishitsu and taijin kyofusho. It is suggested that taijin kyofusho is an excellent example of a mental disorder in which understanding its treatment is an integral part of its conceptualization. PMID- 10454168 TI - The use of Japanese herbal medicine in the treatment of medically unexplained physical symptoms. PMID- 10454169 TI - Rethinking somatoform disorders. PMID- 10454170 TI - To be a victim and a survivor of the great Hanshin-Awaji earthquake. PMID- 10454171 TI - Somatized mental disorder among primary care Arab patients: I. Prevalence and clinical and sociodemographic characteristics. AB - The aims of this study were to estimate the prevalence of somatized mental disorder (SMD) in comparison to psychologized mental disorder (PMD) among a sample of primary health care (PHC) Arab patients, and to investigate the clinical and sociodemographic characteristics of SMD. The first stage of study was conducted by general practitioners (GPs), using the 12-item General Health Questionnaire (GHQ-12). The second stage was carried out by a psychiatrist using the Clinical Interview Schedule (CIS) and an inquiry schedule. Specific operational criteria were used to identify SMD and PMD. The estimated prevalence rate of SMD among the total screened sample was 12%. SMD patients constituted 48% of the psychiatric patients identified, whereas, for the PMD group, this figure was 42%. Educational level was significantly lower in the SMD group. Headache, backache, and abdominal pain were the most commonly presented somatic symptoms. The symptoms pursued a chronic and persistent course and most patients experienced multiple symptoms. The most common ICD-10 psychiatric diagnoses among both SMD and PMD patients were mixed anxiety and depressive disorder, generalized anxiety disorder, and mood and adjustment disorders. Recurrent depressive disorder and dysthymia were significantly more prevalent in the PMD group. The severity of psychiatric illness identified was greater among psychologizers than the somatizers of mental disorder. PMID- 10454172 TI - Development of a Chinese chronic illness-related stress inventory for primary care. AB - This study evaluates the construct validity and internal consistency of a Chronic Illness-Related Stress Inventory (CRSI) for primary care Chinese patients and studies the relationship between chronic illness-related stress and sociodemographic characteristics and indices of disease severity. A total of 301 patients were interviewed using a structured questionnaire. The responses to the CRSI were divided into a frequency scale and a severity scale. Six factors including physical integrity and discomfort, psychosocial function and economic burdens, self-fulfillment and daily life, sexual function, self-esteem, and diet limitations were obtained for each CRSI scale. Except for the last factor, all other factors and the scale as a whole for both scales have a Cronbach alpha of > 0.90. The results of the convergent and discriminant validity analysis were promising. In addition, chronic illness-related stress was related to insurance status and the self-perceived severity of disease. PMID- 10454173 TI - Natural killer (NK) cell activity and NK cell subsets in workers with a tendency of burnout. AB - The involvement of cellular immunity in the burnout syndrome remains to be elucidated. We assessed three components of burnout of the Maslach Burnout Inventory: emotional exhaustion; depersonalization (DP); and personal accomplishment, as well as natural killer cell activity (NKCA) and NK cell subsets in 42 male workers. Workers with a higher DP score showed a lower NKCA and a lower proportionality of CD57+CD16+ to total lymphocytes. There were no differences in any of the health behaviors (e.g., smoking, alcohol, or obesity) between workers showing higher burnout and those showing lower burnout. A stepwise multiple regressions analysis demonstrated that NKCA was closely correlated with DP, independent of other variables, including a stress index. These results suggest that the relationship between reduced cellular immunity and DP is not due to traditional work stress or health behavioral problems. Further studies on DP as a psychosomatic disorder as well as an occupational health problem should be performed in the future. PMID- 10454174 TI - Sympathetic activity in alexithymics with mother's low care. AB - This study examines the relationships between alexithymic characteristics and their psychological factors (i.e., maternal closeness) and the sympathetic system in a sample of male college students. At the resting period, low frequency (LF)/ high frequency (HF) ratio, as an index of sympathetic activity, was significantly higher for students with high scores on global alexithymia or its alexithymia factor (difficulty describing feelings) than those with low scores, suggesting that alexithymic students tend to indicate high sympathetic activity during the resting period. During stress, the increase of LF/HF ratio was higher for students with low scores on alexithymia than those with high scores. Although no significant differences were noted, one may speculate that the differences in stress-related activation of LF/HF are due to higher levels of LH/HF in high alexithymics prior to stress. Our findings indicate that significant association between alexithymia and sympathetic activity during the resting period was controlled for the level of mother's care, suggesting that maternal attachment in infancy and/or childhood may play an important role in the development of alexithymic characteristics and/or sympathetic activity during the resting period. PMID- 10454175 TI - Chronic burnout, somatic arousal and elevated salivary cortisol levels. AB - Burnout syndrome, comprised of the symptoms of emotional exhaustion, physical fatigue, and cognitive weariness, is believed to be a result of ineffective coping with enduring stress. This study of 111 nonshift blue-collar workers free of cardiovascular disease (CVD) examined whether chronic burnout is associated with a state of somatic and physiological hyperarousal. Results showed that 37 workers exhibited symptoms of chronic burnout, with symptoms lasting at least 6 months. These workers, compared to those with no burnout symptoms (n = 52) or nonchronic burnout symptoms (n = 22), had higher levels of tension at work, postwork irritability, more sleep disturbances and complaints of waking up exhausted, and higher cortisol levels during the work day. These results suggest that chronic burnout is associated with heightened somatic arousal and elevated salivary cortisol levels. This may be part of the mechanism underlying the emerging association between burnout and risk of CVD. PMID- 10454176 TI - Syndromic management for sexually transmitted diseases: back to the drawing board. PMID- 10454177 TI - Recurrent vulvovaginal candidiasis: why does it occur? PMID- 10454178 TI - Mucinase and sialidase activity of the vaginal microflora: implications for the pathogenesis of preterm labour. AB - Evidence linking bacterial vaginosis (BV) to chorioamnionitis and spontaneous preterm birth is mounting. Successful treatment of BV could reduce the rate of late miscarriage and preterm birth. Mucinase and sialidase activity have been implicated in the pathogenesis of BV. This study extends the work of previous studies to investigate sialidase, other known mucin degrading enzymes and overall mucin degrading activity in samples of vaginal fluid from women with and without BV. Samples from 31 women were diagnosed for BV, and tested for enzyme activity using established assays. Activity was recorded in all samples. Significant increases in activity were detected in BV samples for sialidase using a mucin (BSM P<0.005) and serum type glycoprotein (AGP P<0.005) substrates, beta galactosidase (P<0.001), and beta-N-acetylhexosaminidase (P<0.01). No significant increases in BV patients were detected in O-glycanase, proteinase, arylesterase, sulphatase or whole mucinase activities. These results support the hypothesis that certain BV-associated enzymes may detrimentally affect the mucosal barrier, permitting bacteria access to the uterus. PMID- 10454179 TI - The rate of diagnosis and demography of pelvic inflammatory disease in general practice: England and Wales. AB - Knowledge of pelvic inflammatory disease (PID) epidemiology is essential to the understanding of reproductive morbidity in women. This paper estimates the rate of PID diagnosis in general practice (GP) and the level of association between PID diagnosis and demographic factors. Diagnoses of PID were made at 1.7% of attendances amongst women aged 16 to 46. Increased risk of PID was associated with smoking (P<0.0001), younger age groups (P<0.0001) and lower socioeconomic groups (P<0.0001). Compared to patients who were married, increased risk was also associated with those patients who were widowed, separated or divorced and not cohabiting (adjusted rate ratio (RR)=1.62; confidence limits (CL) 1.35 to 1.97), and with those who were unmarried but cohabiting (adjusted RR=1.32; 95% CL 1.11 to 1.56). General practice is an important focus for the diagnosis and treatment of PID. If intervention and surveillance are to be undertaken effectively, more has to be known about the epidemiology of this important public health problem. PMID- 10454180 TI - Education attainment as a predictor of HIV risk in rural Uganda: results from a population-based study. AB - We examined the association between education and prevalent HIV-1 infection in the Rakai district, rural Uganda based on a cross-sectional analysis of a population-based cohort. In 1990, 1397 men and 1705 women aged 13 years and older, were enrolled in 31 randomly selected communities. Strata were comprised of main road trading centres, secondary road trading villages and rural villages. Sociodemographic and behavioural data were obtained by interview and serum for HIV serostatus were obtained in the home. The analysis examines the association between sex-specific prevalent HIV infection and educational attainment, categorized as secondary, primary or none. The odds ratios (ORs) and 95% confidence intervals (95% CIs) of HIV infection were estimated, using no education as the referent group. Higher levels of education were associated with a higher HIV seroprevalence in bivariate analyses (OR 2.7 for primary and 4.1 for secondary education, relative to no education). The strength of the association was diminished but remained statistically significant after multivariate adjustment for sociodemographic and behavioural variables (adjusted OR of HIV infection 1.6 (95% CI: 1.2-2.1)) for primary education and 1.5 (95% CI: 1.0-2.2) for secondary education. Stratified multivariate analyses by place of residence indicated that the association between education and HIV prevalence was statistically significant in the rural villages, but not in the main road trading centres and intermediate trading villages. Educational attainment is a significant predictor of HIV risk in rural Uganda, in part because of risk behaviours and other characteristics among better educated individuals. Preventive interventions need to focus on better educated adults and on school aged populations. PMID- 10454181 TI - Reactivity of a dual amplified chlamydia immunoassay with different serovars of Chlamydia trachomatis. AB - A study was undertaken with different serovars (D, E, F, L2, MoPn) of Chlamydia trachomatis to determine the analytical sensitivity of a new dual amplified immunoassay (IDEIA PCE Chlamydia) for detecting chlamydial lipopolysaccharide. IDEIA PCE Chlamydia incorporates a polymer conjugate consisting of multiple copies of antibody and enzyme molecules to provide signal amplification. The test was also assessed with different protein A producing strains of Staphylococcus aureus in order to assess whether the use of a multiple antibody conjugate increased nonspecific binding. The detection limits varied for each serovar with a detection limit of 38 IFU/ml obtained with serovar F and 237 IFU/ml obtained with serovar D. The incorporation of the polymer conjugate resulted in a 2-5 fold increase in analytical sensitivity compared to an earlier version of the test using a conventional conjugate. No increase in cross reactivity with protein A producing strains of S. aureus was obtained. The new dual amplified test format offers potential as a sensitive low-cost screening assay for C. trachomatis infections. PMID- 10454182 TI - Survey of risk behaviour and HIV prevalence in an English prison. AB - An anonymous, voluntary, linked cohort study was undertaken to determine the prevalence of HIV infection and identify risk factors for the spread of infection in an English prison. Three hundred and seventy-eight (68%) of the inmates participated. The HIV point prevalence was 0.26%. Injecting drug use (IDU) was the most significant HIV risk factor within 20% admitting IDU at any time, of whom 58% injected whilst in prison. Of those injecting in prison 73% shared needles. Two inmates admitted having sex with a male partner in prison. This study demonstrates that the potential exists in this setting for an outbreak of blood-borne virus infection; hepatitis B virus (HBV), hepatitis C virus (HCV) and HIV infection. Injecting drug use and needle sharing represent the greatest risk. PMID- 10454183 TI - Immunological predictors of HIV-related survival. AB - While the prognostic role of CD4 lymphocyte count is well established, little is known about how other immunological factors are associated with HIV-related survivorship. CD8 lymphocyte count is reported to inhibit growth of HIV in vitro and to deter the development of cytomeglovirus infections. Scarce information exists, however, on its association with HIV-related survivorship. The objective of the present study therefore was to assess whether survival estimation could be improved by including CD8 lymphocyte count as a prognostic factor. The study sample consisted of 904 adult HIV-infected patients who were treated between 1992 and 1997 at a large southwestern academic medical centre in the US. Survival analysis using proportional hazards modelling showed that baseline CD4 lymphocyte was highly and positively predictive of HIV-related survival. By contrast, baseline CD8 count was not predictive of survival in the study cohort. While the interaction between CD4 and CD8 was not statistically significant in the present study sample, decomposition of this effect indicates that these factors may operate synergistically to predict survival. In conclusion, these findings suggest that clinicians may improve their prognostic accuracy by relying on both CD4 and CD8 lymphocyte counts rather than relying exclusively on CD4 counts. PMID- 10454184 TI - HIV/AIDS in Bangladesh: a national surveillance. AB - Nationwide surveillance of HIV/AIDS from 1989 through 1996 in Bangladesh included several risk groups such as professional blood donors, patients with sexually transmitted diseases (STDs), pregnant women at antenatal clinics, commercial sex workers (CSWs), patients with tuberculosis, long-distance truck drivers, sailors, and non-residents. The population was enrolled by convenience sampling after taking informed consent. Among 70,676 persons tested, 80 (1.13 per 1000) were HIV positive. The prevalence rate was steady until 1994, and then increased rapidly. The rate among male heterosexuals was significantly higher than that in females (3.40 per 1000 versus 0.29 per 1000; odds ratio (OR) 11.60; 95% confidence intervals (CI) 6.45 to 21.16; P < 0.0001). Twelve per cent of patients with STDs had HIV. The HIV cases concentrated in 2 districts, Sylhet (25/72) and Chittagong (20/72), that border India and Myanmar (formerly Burma), respectively. Frequent movement of people of Bangladesh to India, Pakistan, Myanmar and Thailand, where HIV rates are higher, is one of the possible sources of spread of the cases. Bangladesh has the potential to avert epidemic spread of HIV at its early stage. PMID- 10454185 TI - High prevalence of sexually transmitted and blood-borne infections amongst the inmates of a district jail in Northern India. AB - Two hundred and forty male and 9 female jail inmates confined for various crimes in a district jail near Delhi were screened for sexually transmitted and blood borne diseases including HIV, syphilis and hepatitis B and C viral infections, skin diseases etc. The inmates were aged 15-50 years with a mean of 24.8+/-0.11. Their alleged criminal background, period of stay in the jail, drug addiction, education, birth place, marital status, sexual activity, and clinical complaints were recorded by an anonymous questionnaire. Serum samples were tested for antibodies against HIV (1+2), hepatitis C (HCV), Treponema pallidum and for hepatitis B surface antigen (HBsAg). Sputum examination was done for acid-fast bacilli. Out of the 240 men, 115 were married and 125 unmarried. One hundred and eighty-four (76.6%) men gave history of penetrative sex. Of the 184, 53 (28.8%) were homosexuals or bisexuals and 131 (71.2%) had sex with women only. Sixty of the 131 (45.8%) were faithful to their partners while 124 gave a history of having multiple sexual partners and 100 of them (80.6%) had unprotected sex. Eighty-three of these 100 also had had sex with commercial sex workers (CSWs). One hundred and twenty-six were addicted for alcohol, 44 for smack/charas and 8 had a history of intravenous drug abuse. One hundred and seventy-four were not aware of AIDS. On examination 28 of the 240 (11.6%) had active hepatitis with or without a history of jaundice in the last 2 years, 25 (10.4%) active pulmonary tuberculosis (TB) and 11 (4.6%) had syphilitic ulcers on the penis. Four-fifths of the teenagers confined to a particular barrack had moderate to severe scabies. Three males (1.3%) were found to be Western blot confirmed HIV-1 positive while 28 (11.1%) men and 2 (22.2%) women were positive for HBsAg. Twelve (5.0%) men but no women, were found to be positive for anti-HCV antibodies. Out of the 3 HIV positive persons, one was an intravenous drug user (IVDU), second was a drug addict and frequent CSW visitor while the third was a homosexual. This pilot study gives an indication that sexually transmitted and blood-borne infections are highly prevalent in jail premises and pose a threat of rapid spread of these infections through IVDU and homosexuality. PMID- 10454186 TI - Blue nails are a sign of HIV infection. PMID- 10454187 TI - Ocular and fulminant cerebral lymphoma in AIDS. PMID- 10454188 TI - Role of HTLV-1 co-infection in the AIDS epidemic in the Caribbean: a cause for concern. PMID- 10454189 TI - An audit of chlamydia treatment and contact tracing in a sexual health service. PMID- 10454190 TI - Protein phosphorylation and signal transduction. AB - It is now generally accepted that protein phosphorylation-dephosphorylation has a role in the regulation of essentially all cellular functions. Thus, it is of interest that this process is involved in signal transduction. Nonetheless, the extent to which protein phosphorylation participates in signaling is truly remarkable. Almost every known signaling pathway eventually impinges on a protein kinase, or in some instances, a protein phosphatase. The diversity of these enzymes is noteworthy, and it is of interest that many biotechnology companies are eyeing them as potentially important targets for drugs. Such drugs may have important therapeutic applications, and in any event, they certainly will be useful to investigators who study signal transduction. Indeed, this already has been proven to be true. PMID- 10454191 TI - H-series protein kinase inhibitors and potential clinical applications. AB - During the course of generating derivatives of N-(6-aminohexyl)-5-chloro-1 naphthalene-sulfonamide, a synthetic calmodulin inhibitor, we came across several analogues with shorter alkyl chains that exhibited inhibition of serine/threonine protein kinase activities in an ATP-competitive manner. Certain derivatives proved to be selective inhibitors of protein kinases useful for elucidation of relevant functions of the enzymes. One of them turned out to be a unique vasodilator that preferentially suppresses delayed cerebral vasospasm, a critical complication of subarachnoid hemorrhage, without significant changes in systemic blood pressure. The compound in question, 1-(5-isoquinolinesulfonyl) homopiperazine, was identified from sequential development of protein kinase inhibitors with isoquinolinesulfonyl structures, which occupy the adenine pocket of the ATP-binding site of the enzyme. It recently has been proposed that the target kinase responsible for vasodilation by 1-(5-isoquinolinesulfonyl) homopiperazine may be Rho-kinase, which regulates phosphorylation of myosin light chains and vasocontraction. Because protein phosphorylation plays important roles in regulation of various cellular functions, the foregoing is a good example of current progress in the development of protein kinase inhibitors with potential clinical applications. PMID- 10454192 TI - Catalytic subunit of cyclic AMP-dependent protein kinase: structure and dynamics of the active site cleft. AB - The catalytic subunit of cyclic AMP-dependent protein kinase serves as a structural template for the entire family of Ser, Thr, and Tyr specific protein kinases. We review here the dynamics of the active catalytic subunit. These dynamics correlate with an opening and closing of the active site cleft, and are considered to be a requirement for catalysis. The motions, described by a set of several crystal structures, reveal a very fluid active site cleft. This active site cleft with its dynamic opening and closing is a prime target for protein kinase inhibitors. PMID- 10454193 TI - Substrate and inhibitor recognition of protein kinases: what is known about the catalytic subunit of phosphorylase kinase? AB - Although much can be learned about the specificity of protein kinases from studies with peptide substrates, the question remains, how do kinases recognize their three-dimensional protein substrates? Information derived from such studies provides further understanding of substrate recognition and can facilitate the design of specific protein kinase inhibitors. Phosphorylase kinase (PhK) catalyzes the phosphorylation of phosphorylase b (phos. b) to form the active phosphorylase a. No other protein kinase can duplicate this reaction. Why? To probe this question and establish what features in the protein are important for substrate binding and product release, mutants of phos. b have been studied. This report shows how mutations change the properties of the protein substrate and the ability of these mutants to be phosphorylated by PhK and other kinases. Action of protein kinases on their substrates is often regulated by autoinhibitory segments. The C-terminus of the catalytic gamma-subunit of PhK contains two inhibitory sites overlapping two calmodulin-binding regions. These two peptide segments resemble sequences in phos. b and may explain why peptides of these regions are potent inhibitors of PhK. We will show results with peptide inhibitors, using various expressed forms of the catalytic subunit, which describe their modes of interaction and mechanisms of inhibition. Metal ions can change molecular interactions. With PhK, Mn2+ facilitates the use of GTP as a phosphoryl group donor and greatly increases phosphorylation of a tyrosine residue in angiotensin II. This implies that the spatial arrangement of specificity determinants can be manipulated so that PhK can utilize other substrates. PMID- 10454194 TI - Conformational diversity of catalytic cores of protein kinases. AB - X-ray crystallography of the protein kinase family has provided an impressive array of crystal structures, setting the stage for rational design of specific inhibitors of these vitally important regulators of the signaling pathways of the cell. Initial work on the first crystal structure of a protein kinase, cyclic AMP dependent protein kinase, has provided evidence of conformational changes suggested to be critical for the common catalytic event of transferring the gamma phosphate from ATP onto the targeted protein. This review updates the current status of the extent of conformational diversity of the protein kinase family and suggests that both the nature and the extent of those changes can provide a rationale for the increased occurrence of specific protein kinase inhibitors targeted at the ATP-binding site. It focuses on the fact that in addition to the sequence diversities in ATP binding clefts reported recently, there is conformational diversity in the beta sheets of the upper domains of the catalytic cores. This difference is directly related to the regulation of kinases by multiple mechanisms. PMID- 10454195 TI - KID, a Kinase Inhibitor Database project. AB - The Kinase Inhibitor Database is a small specialized database dedicated to the gathering of information on protein kinase inhibitors. The database is accessible through the World Wide Web system and gives access to structural and bibliographic information on protein kinase inhibitors. The data in the database will be collected and submitted by researchers working in the kinase inhibitor field. The submitted data will be checked by the curator of the database before entry. PMID- 10454196 TI - Small molecule inhibitors of tumor-promoted angiogenesis, including protein tyrosine kinase inhibitors. AB - Angiogenesis is an exciting and promising new area of research. The concept that tumor cells are absolutely dependent upon neovascularization to grow and metastasize has opened the door to a multitude of new approaches and targets for developing anticancer therapies. These potential new antiangiogenic therapies offer the possibility for improved efficacy and reduced toxicity relative to conventional cancer treatments without the possibility of drug resistance. In particular, reports of small molecule inhibitors of tumor-promoted angiogenesis are appearing ever more frequently in the scientific literature. For this reason, the major focus of this review will be to cover small molecule inhibitors of tumor-promoted angiogenesis. The present review concentrates on selected literature, principally from mid-1996 to mid-1998, where there are sufficient biological data to support claims of antiangiogenic activities by small molecules. In addition, a historical background is presented and some of the important issues related to this field are discussed within. PMID- 10454197 TI - Strategies toward the design of novel and selective protein tyrosine kinase inhibitors. AB - Protein tyrosine kinases play a fundamental role in signal transduction pathways. Deregulated tyrosine kinase activity has been observed in many proliferative diseases (e.g., cancer, psoriasis, restenosis, etc.). Tyrosine kinases are, therefore, attractive targets for the design of new therapeutic agents against cancer. We have built up a pharmacophore model of the ATP-binding site of the epidermal growth factor receptor (EGFR) kinase and used it for the rational design of kinase inhibitors. Several examples of the successful use of this model are presented in this review. Amongst these, 4-substituted-pyrrolo[2,3 d]pyrimidines, a new class of highly potent and selective inhibitors of the EGFR kinase, have been identified and further optimized. The most active derivatives inhibited the EGFR tyrosine kinase with IC50 values between 1 and 5 nM. In EGF dependent cellular systems, tyrosine phosphorylation, as well as c-fos mRNA expression, was inhibited with similar IC50 values. Further successful application of this pharmacophore model led to the identification and optimization of phenylamino-pyrazolo[4,3-d]pyrimidines and substituted isoflavones and quinolones, other classes of potent, selective, and ATP competitive EGFR kinase inhibitors with IC50 values in the low nanomolar range. Structure-activity relationships of both classes are discussed. PMID- 10454198 TI - Inhibition of the epidermal growth factor receptor family of tyrosine kinases as an approach to cancer chemotherapy: progression from reversible to irreversible inhibitors. AB - The rationale to inhibit the epidermal growth factor receptor (EGFR) tyrosine kinase family as an approach to cancer chemotherapy has continued to grow stronger over the last 10 years. Both preclinical and clinical data strongly support the involvement of these receptors in the formation and progression of human cancers, as well as establish a high correlation in cancer patients between receptor/ ligand expression and poor prognosis. During the past 4 years, significant progress has been made in the area of EGFR tyrosine kinase inhibitors, and new structural classes have emerged that exhibit enormous improvements with regard to potency, specificity, and in vitro and in vivo activity. Very recently, further advancements in this field have been made whereby very specific, irreversible inhibitors of the EGFR family have been synthesized that provide unique pharmacological properties and exceptional efficacy. The in vivo performance of these modern kinase inhibitors has improved to the point where several compounds are either in clinical trials or very near to that point in their development. This review will briefly address the justification for targeting the EGFR family for cancer therapeutics, and then will highlight some of the more promising kinase inhibitors that are in development. PMID- 10454199 TI - Chemical approaches to the study of protein tyrosine kinases and their implications for mechanism and inhibitor design. AB - Protein tyrosine kinases are critical enzymes for signal transduction. Using C terminal Src kinase (Csk) as a model system, we discuss progress in three main areas. First, we describe our efforts to measure the transition state of the reaction using peptide substrates containing fluorotyrosine analogs. It is shown that the Bronsted nucleophile coefficient for the reaction is near zero (similar to the nonenzymatic reaction) and the required nucleophile is the neutral phenol (rather than the more chemically reactive phenoxide anion). By studying the kinase reaction in the reverse direction, a Bronsted leaving group coefficient of -0.3 was measured, indicative of protonation of the departing phenol in the transition state. Taken together, these results strongly support a dissociative transition state mechanism for the kinase. These findings set constraints on the design of transition state analog inhibitors. Second, we describe efforts toward defining the specificity of Csk for peptide and protein substrates. The main findings are that local amino acids surrounding a phosphorylated tyrosine can influence recognition, but that long-range interactions probably are more important in a physiologic protein substrate. These findings underscore the complexities in how protein kinases select protein substrates. Third, we describe a new method in protein engineering that has been applied to the study of protein kinases. The method, expressed protein ligation, allows a general approach for ligating synthetic peptides to recombinant proteins. Using expressed protein ligation, obtaining site-specifically phosphorylated proteins and proteins with the incorporation of biophysical probes becomes relatively straightforward. We have used this method to generate a tail phosphorylated, conformationally altered Csk that showed an unexpected increase in kinase activity. PMID- 10454200 TI - Protein tyrosine kinase inhibitors as novel therapeutic agents. AB - Protein tyrosine kinases (PTKs) play a key role in normal cell and tissue development. Enhanced PTK activity is intimately correlated with proliferative diseases, such as cancers, leukemias, psoriasis, and restenosis. This realization prompted us to systematically synthesize tyrosine phosphorylation inhibitors (tyrphostins) as potential drugs. Over the years, we have demonstrated the ability to synthesize selective tyrphostins aimed at different receptor, as well as at nonreceptor, tyrosine kinases. Some of these tyrphostins have shown efficacy in vivo as antileukemic agents and antirestenosis agents. AG 490, a Jak 2 inhibitor, is potent against recurrent pre-B acute lymphoblastic leukemia. AG 1295, a selective platelet-derived growth factor receptor kinase inhibitor, inhibits 50% of balloon injury-induced stenosis in the phemoral arteries of pigs. AG 1517 (SU 5271), a potent epiderminal growth factor receptor kinase inhibitor, is currently in clinical trials for psoriasis. Similarly, SU 5416, a potent kinase inhibitor of the vascular endothelial growth factor receptor/kinase domain receptor/Flk-1, is currently in clinical trials as an anticancer agent by virtue of its strong anti-angiogenic activity. These findings demonstrate that the identification of PTKs that play a key role in a defined disease state can lead to a selective drug. Tyrphostins also show efficacy in vivo in inflammatory diseases such as sepsis, cirrhosis, and experimental autoimmune encephalitis. PMID- 10454201 TI - The epidermal growth factor receptor and its inhibition in cancer therapy. AB - Much effort has been expended in the search for inhibitors of signalling molecules that may prove to be important therapeutically in cancer. The epidermal growth factor receptor (EGFR) family and their associated ligands has been one such area extensively investigated. The complex nature of EGFR biology allows for potential opportunities for EGFR inhibitors in a number of areas of cancer therapy, including proliferative, angiogenic, invasive, and metastatic aspects. Much positive evidence of likely benefit has already been gathered from a multiplicity of laboratory experiments. Clinical trials are now urgently required to further evaluate the advantages of such agents. PMID- 10454203 TI - Inhibition of protein kinase C--do we, can we, and should we? AB - The second messenger responsive protein kinase C (PKC) family has promised much as therapeutic targets. Indeed, there has been excellent progress in the development of highly selective inhibitors for this class of kinases. The key to the further development and use of inhibitors selective within the family depends upon our understanding of the involvement of PKC in human disease. Recent work in model systems ex vivo has modified the simplistic view of PKC control and broadened the spectrum of signalling pathway involvement. The consequence is that there is a need to reassess the role of PKC that to date has coloured our view of PKC and disease. These developments, their implications, and the future assessment of PKC as a target are discussed. PMID- 10454202 TI - The transition from a pharmacophore-guided approach to a receptor-guided approach in the design of potent protein kinase C ligands. AB - The pharmacophore-guided approach used in the first phase of the design of novel protein kinase C (PKC) ligands was based on the study of the geometry of bioequivalent pharmacophores present in diacylglycerol (DAG) and in the more potent phorbol ester tumor promoters. A number of potent DAG lactones were generated by this approach, in which the glycerol backbone was constrained into various heterocyclic rings to reduce the entropic penalty associated with DAG binding. Based on the information provided by X-ray and NMR structures of the cysteine-rich, C1 phorbol ester/DAG binding domain, the DAG lactones were further modified to optimize their interaction with a group of highly conserved hydrophobic amino acids along the rim of the C1 domain. This receptor-guided approach culminated with the synthesis of a series of "super DAG" molecules that can bind to PKC with low nanomolar affinities. These compounds provide insight into the basis for PKC ligand specificity. Moreover, some of the compounds reviewed herein show potential utility as antitumor agents. PMID- 10454204 TI - Chemical inhibitors of cyclin-dependent kinases: insights into design from X-ray crystallographic studies. AB - Cyclin-dependent kinases (CDKs) are a family of protein kinases that regulate progression through the eukaryotic cell cycle. Aberrant CDK activity or function is a common defect in human tumours, resulting in unrestrained cellular proliferation. X-ray crystallographic analysis of monomeric CDK2 and CDK2 complexes has revealed how phosphorylation and cyclin binding mediate enzyme activation and how this activity can be regulated by further protein association. Current research aims to improve the selectivity and/or potency of small molecule CDK inhibitors, both to develop specific probes to study the roles of the different CDK family members in coordinating cell cycle progression, and as lead molecules for the design of therapeutically useful drugs. This design process has been assisted by the availability of a number of CDK2/inhibitor structures determined using X-ray crystallography. These structures have shown that molecules related to ATP can be accommodated in the ATP-binding site in a number of orientations, utilising interactions observed between CDK2 and its natural ligand, as well as novel interactions with CDK2 residues that lie both within and outside the active site cleft. This site can also bind inhibitors that are structurally unrelated to ATP. These results suggest that it may be possible to design pharmacologically and pharmaceutically important ATP-binding site-directed ligands that act as specific and potent inhibitors of CDK activity. PMID- 10454205 TI - Properties and potential-applications of chemical inhibitors of cyclin-dependent kinases. AB - Cyclin-dependent kinases (CDKs) trigger and co-ordinate the cell division cycle phases. They also play a role in neuronal cells and in the control of transcription. Intensive screening has led in the past few years to the identification of a series of chemical inhibitors of CDKs. Some of these compounds display remarkable selectivity and efficiency (IC50 <25 nM). Many have been co-crystallised with CDK2, and their atomic interactions with the kinase have been analysed in detail: all are located in the ATP-binding pocket of the enzyme. These inhibitors are antimitotic, they arrest cells in G1 and, at higher doses, in G2/M. Furthermore, they facilitate or even trigger apoptosis in proliferating cells. In contrast, they protect neuronal cells from apoptosis. The potential use of these inhibitors is being extensively evaluated in cancer chemotherapy (clinical trials, Phase I and II). Possible clinical applications are being investigated in other fields: cardiovascular (restenosis, tumoural angiogenesis, atherosclerosis), nephrology (glomerulonephritis), dermatology (psoriasis), parasitology (unicellular parasites such as Plasmodium, Trypanosoma, Toxoplasma, etc.), neurology (Alzheimer's disease), viral infections (cytomegalovirus, human immunodeficiency virus, herpes). We anticipate the discovery of novel selective and powerful inhibitors in the near future, and hope for their efficient applications in various human diseases. PMID- 10454206 TI - Cyclin-dependent kinases: initial approaches to exploit a novel therapeutic target. AB - Cyclin-dependent kinases (CDKs) have been recognized as key regulators of cell cycle progression. Alteration and deregulation of CDK activity are pathogenic hallmarks of neoplasia. Therefore, inhibitors or modulators would be of interest to explore as novel therapeutic agents in cancer, as well as other hyperproliferative disorders. Flavopiridol is a semisynthetic flavonoid that emerged from an empirical screening program as a potent antiproliferative agent that mechanistic studies demonstrated to directly inhibit CDKs 1, 2, and 4 as a competitive ATP site antagonist. Initial clinical trials have shown that concentrations that inhibit cell proliferation and CDK activity in vitro can be safely achieved in humans, and additional clinical trials will establish its clinical potential. To address the need for additional chemotypes that may serve as lead structures for drugs that would not have the toxicities associated with flavopiridol, compounds with a similar pattern of cell growth inhibitory activity in the National Cancer Institute's in vitro anticancer drug screen have been recognized by the computer-assisted pattern recognition algorithm COMPARE and then screened for anti-CDK activity in a biochemical screen. The benzodiazepine derivative NSC 664704 (7,12-dihydro-indolo[3,2-d][1]benzazepin-6(5H)-one) was revealed by that approach as a moderately potent (IC50 0.4 microM) inhibitor of CDK2, which in initial experiments shows evidence of causing cell cycle redistribution in living cells. NSC 664704 is, therefore, a candidate for further structural optimization, guided in part by understanding of the ATP-binding site in CDK2. This approach represents one way of combining empirical screening information with structure-based design to derive novel candidate therapeutic agents directed against an important cellular target. PMID- 10454207 TI - Inhibitors of protein kinases: CGP 41251, a protein kinase inhibitor with potential as an anticancer agent. AB - CGP 41251 was originally identified as an inhibitor of protein kinase C (PKC), inhibiting mainly the conventional PKC subtypes, and subsequently shown to inhibit the vascular endothelial growth factor (VEGF) receptor kinase insert domain-containing receptor, which is involved in angiogenesis. CGP 41251 inhibits reversibly intracellular PKC activity, induction of c-fos and the corresponding activation of the mitogen-activated protein kinase induced by either tumor promoting phorbol esters, platelet-derived growth factor, or basic fibroblast growth factor, but not by the epidermal growth factor. CGP 41251 inhibited the ligand-induced autophosphorylation of the receptors for platelet-derived growth factor, stem cell factor, and VEGF (kinase insert domain-containing receptor) that correlated with the inhibition of the mitogen-activated protein kinase activation, but did not affect the ligand-induced autophosphorylation of the receptors for insulin, insulin-like growth factor-I, or epidermal growth factor. CGP 41251 showed broad antiproliferative activity against various tumor and normal cell lines in vitro, and is able to reverse the p-glycoprotein-mediated multidrug resistance of tumor cells in vitro. CGP 41251 showed in vivo antitumor activity as single agent and inhibited angiogenesis in vivo. Thus, CGP 41251 may suppress tumor growth by inhibiting tumor angiogenesis (via its effects on the VEGF receptor tyrosine kinases) in addition to directly inhibiting tumor cell proliferation (via its effects on PKCs). PMID- 10454208 TI - CK2: a protein kinase in need of control. AB - Protein kinase CK2 is a heterotetrameric alpha2beta2 Ser/Thr protein kinase with some features unusual among the eukaryotic protein kinases: (1) CK2 recognizes phosphoacceptor sites specified by several acidic determinants; (2) CK2 can use both ATP and GTP as phosphoryl donors; and (3) the regulatory properties of CK2 are poorly understood; it is insensitive to any known second messenger and displays high basal activity. To gain insight into CK2 regulation and to understand its unusual properties, site-directed mutagenesis experiments on both subunits and X-ray crystallographic studies of the catalytic alpha-subunit were performed. The noncatalytic beta-subunit has at least three functions: (1) it protects the alpha-subunit against denaturing agents or conditions; (2) it alters the substrate specificity of the alpha-subunit; and (3) it modulates the activity of the enzyme, i.e., depending on the substrate, it increases or decreases the activity of the alpha-subunit. Mutagenesis experiments revealed that an acidic stretch between amino acids 55 and 64 has a down-regulatory and autoinhibitory function. Mutational analysis of the alpha-subunit has revealed a network of unique basic residues that are responsible for the recognition of phosphoacceptor substrates and for down-regulation by the beta-subunit and by polyanionic inhibitors. The resolution of the crystal structure of Zea mays CK2 alpha-subunit has disclosed the structural features that are responsible for high basal activity and for unusual response to nucleotide analogs. The increasing knowledge of CK2 structure-function relationships will allow the design of highly selective inhibitors of this pleiotropic kinase with oncogenic potential. PMID- 10454209 TI - Viral and host-cell protein kinases: enticing antiviral targets and relevance of nucleoside, and viral thymidine, kinases. AB - Numerous targets are known for development of antiviral agents, and some significant successes have been achieved with nucleoside analogues. These are "activated" by phosphorylation by viral and/or host-cell nucleoside kinases, the final target being principally the viral polymerase. With latency of herpes viruses, the viral thymidine kinase may be the ultimate target. Less attention has been devoted to viral protein kinases as antiviral targets, largely because 5 years ago, these the study of such enzymes was considered "still in its infancy." In the interim, identification of viral and host-cell protein kinases involved in viral gene expression, and viral replication, has made impressive advances. In conjunction with current progress in development of specific inhibitors of cellular protein kinases, and the differences in sequence motifs between these and the viral enzymes, the latter are indeed attractive targets, as are also some host-cell protein kinases. Examples include, amongst others, the essential protein kinases of vaccinia virus; the nonsegmented negative-strand RNA viruses, all essentially dependent on host-cell kinases, e.g., protein kinase CK-II (casein kinase-II), for which good inhibitors, such as halogenated benzimidazoles and benzotriazoles, are known; herpes viruses, with emphasis on human cytomegalovirus, the UL97 gene of which codes for a protein kinase that, like viral thymidine kinases, "activates," by phosphorylation, a nonpeptide antiviral acyclonucleoside ganciclovir, an analogue of the antiherpes aciclovir. The latter, in turn, is active against animal cytomegaloviruses following phosphorylation by the products of their UL97 gene homologues. Attention is also directed to the antiviral activity of the cyclic phosphate of ganciclovir, a structural analogue of the second messenger cyclic GMP. PMID- 10454210 TI - Acquisition of inhibitor-sensitive protein kinases through protein design. AB - Protein phosphorylation is the major post-translational modification used by eukaryotic cells to control cellular signaling. Protein kinases have emerged as attractive drug targets because heightened protein kinase activity has been associated with several proliferative diseases, most notably cancer and restenosis. Until now, it has been very difficult to confirm the utility of protein kinases as inhibitor targets because very few small molecules that selectively inhibit one particular kinase are known. Discovery of highly specific kinase inhibitors has been slow because the protein family contains approximately 2000 members, all of which share a conserved active site fold. Recent work in several laboratories has sought to circumvent the problem of kinase structural degeneracy by engineering drug sensitivity into Src family tyrosine kinases and mitogen-activated protein kinases through site-directed mutagenesis. By introducing a unique non-naturally occurring amino acid into a conserved region of the enzyme's binding site, a target protein kinase can be rapidly sensitized to a small molecule. Introduction of the engineered kinase into a cell line or animal model should greatly expedite the investigation of protein kinase inhibition as a viable drug treatment. The purpose of this review is to summarize these recent advances in protein kinase drug sensitization. PMID- 10454211 TI - Caged regulators of signaling pathways. AB - Although inhibitors can implicate individual protein kinases in specific signal transduction pathways, simple reversible inhibitors are unable to define both the time of initiation and the duration of protein kinase activity. How long must a specific protein kinase remain active in a particular transduction cascade to ensure that the message results in the appropriate cellular response? What is the precise temporal assembly order of individual components of an intracellular signaling complex? What, if any, temporal crosstalk is there between individual members of bisecting signaling pathways? Caged protein kinase inhibitors, as well as caged protein kinases, should serve as useful tools for temporal studies of signal transduction. We have prepared both a caged cyclic AMP-dependent protein kinase and a caged inhibitor of this enzyme. Upon photolysis, the functional activities of both species are dramatically unleashed in vitro. We have also explored both the activation and the inhibition of the cyclic AMP-dependent protein kinase-driven signaling pathway in rat embryo fibroblasts with these caged species via microinjection, and have now demonstrated that the activity of signal transduction pathways can be controlled by light in living cells. PMID- 10454212 TI - Cyclic GMP-dependent protein kinases: understanding in vivo functions by gene targeting. AB - Identifying the specific functions that are mediated by cyclic GMP (cGMP) dependent protein kinases is key to our understanding of the biological role of the nitric oxide/cGMP signaling cascade. Over the last two decades, there have been numerous reports on the functional roles of cGMP kinases. However, most studies have been performed with isolated cells and organs by using specific activators and inhibitors of cGMP kinases. Under such experimental conditions, a clear dissection between the cGMP kinase pathway and other pathways, for example, that of cyclic AMP kinase, has been difficult to achieve. In order to identify biological processes in which cGMP kinases are unambiguously involved, the genes of cGMP kinase I and cGMP kinase II have been deleted in mice. This review focuses on the physiological functions that are regulated by cGMP kinases as revealed by cGMP kinase-deficient animals. For cGMP kinase I, these functions include the contractility of vascular and gastrointestinal smooth muscle and the homeostasis of platelet activity. According to its expression pattern, the Type II cGMP kinase plays a definite biological part in transepithelial Cl- and Na+ transport in the intestine, longitudinal growth of long bones, and the regulation of the kidney renin-angiotensin-aldosterone system. Further, cGMP kinase I and II mutants reveal new aspects for the role of nitric oxide/cGMP in the induction of neuronal plasticity, such as hippocampal long-term potentiation and cerebellar long-term depression. In conclusion, inactivation of the cGMP kinase genes shows that cGMP kinases regulate very specifically distinct cellular functions by pathways that are separate from those used by cyclic AMP kinases. Mice deficient in cGMP kinases exhibit severe defects and, therefore, may serve as animal models for several human diseases, including hypertension, thrombosis, gastrointestinal dysmotility, and dwarfism. PMID- 10454213 TI - Delineation of selective cyclic GMP-dependent protein kinase Ialpha substrate and inhibitor peptides based on combinatorial peptide libraries on paper. AB - Peptide libraries on cellulose paper have proven to be valuable tools for the a priori determination of substrate specificities of cyclic AMP- and cyclic GMP dependent protein kinases (cAMP-kinase and cGMP-kinase) on the basis of octa peptide sequences. Here, we report the extension of our peptide library screens to 12-mer and 14-mer peptide sequences, resulting in highly cGMP-kinase Ialpha selective peptides. The sequences TQAKRKKSLAMA-amide and TQAKRKKSLAMFLR-amide, with Km values for cGMP-kinase Ialpha of 0.7 and 0.26 microM and Vmax values of 11.5 and 10.9 micromol/min/mg, respectively, display a high specificity for this enzyme. Furthermore, replacing the phosphate acceptor residue serine with alanine in TQAKRKKSLAMA-amide resulted in the highly cGMP-kinase Ialpha selective inhibitor peptide TQAKRKKALAMA-amide, with inhibitor constants for cGMP-kinase Ialpha and cAMP-kinase of 7.5 microM and 750 microM, respectively. Selective cGMP kinase inhibitors have the potential to play an important role in the elucidation of the distinct cellular functions of cGMP-kinase separate from those activated by cAMP-kinases, and, therefore, may play an important role as pharmaceutical targets. Molecular docking experiments of the most cGMP-kinase selective sequences on a molecular model of the catalytic domain of cGMP-kinase Ialpha suggest that they adopt unique conformations, which differ significantly from those observed for the cAMP-kinase-specific inhibitor PKI(5-24). Our results suggest that despite their structural similarities, cAMP-kinase and cGMP-kinase use distinct peptide substrate and inhibitor conformations, which could account for their unique substrate specificities. These findings are further supported by cAMP- and cGMP-kinase-selective inhibitor analogs with (D)-Ala residues at the inhibitory positions. PMID- 10454214 TI - p38 mitogen-activated protein kinase inhibitors--mechanisms and therapeutic potentials. AB - The pyridinylimidazole compounds, exemplified by SB 203580, originally were prepared as inflammatory cytokine synthesis inhibitors. Subsequently, the compounds were found to be selective inhibitors for p38 mitogen-activated protein kinase (MAPK), a member of the MAPK family. SB 203580 inhibits the catalytic activity of p38 MAPK by competitive binding in the ATP pocket. Four homologues of p38 MAPK have been identified to date, and interestingly, their biochemical properties and their respective sensitivities to the inhibitors are distinct. X ray crystallographic analysis of p38-inhibitor complexes reinforces the observations made from site-directed mutagenesis studies, thereby providing a molecular basis for understanding the kinase selectivity of these inhibitors. The p38 MAPK inhibitors are efficacious in several disease models, including inflammation, arthritis and other joint diseases, septic shock, and myocardial injury. PMID- 10454215 TI - Peptide inhibitors of the mitogen-activated protein kinase pathway: a structure mimetic peptide corresponding to the conserved inter-DFG-APE region in the kinase domain. AB - The signal transduction pathway mediated by mitogen-activated protein kinases is an attractive target for the design of pharmacologically effective inhibitors. Two specific cell-permeant small molecule inhibitors of this pathway have been reported. However, under certain circumstances, nonpermeable inhibitors, such as neutralizing antibodies and peptide inhibitors, are also useful. We present here a novel approach for such peptide inhibitor design. The procedure is based on the synthesis of a structure-mimetic peptide corresponding to a short peptide segment in the target molecule. The results obtained so far show that a peptide designed in such a way is an effective inhibitor of the pathway. The possible application of such peptides and antipeptide antibodies as probes for protein kinase regulation mechanisms is also evaluated. PMID- 10454217 TI - Pharmacology of antisense oligonucleotide inhibitors of protein expression. AB - The dramatic increase in recent years of both the amount and rate of accumulation of novel genomic sequence information has generated enormous opportunities for the development of new classes of drugs. For these opportunities to be fully capitalized upon, investigators must choose molecular targets for drug development that are likely to yield attractive therapeutic profiles. This will require rapid and effective determination of gene functions in multiple cellular settings. The development of antisense oligonucleotides as specific inhibitors of gene expression should allow such determination of gene function. In addition, the antisense oligonucleotides themselves will likely prove useful as drugs. In this review, we discuss some of the issues surrounding the use of antisense oligonucleotides as research tools to help elucidate gene function, and highlight some of the approaches that can be taken to generate and use effective antisense reagents. PMID- 10454216 TI - Mechanism of protein kinase B activation by insulin/insulin-like growth factor-1 revealed by specific inhibitors of phosphoinositide 3-kinase--significance for diabetes and cancer. AB - Protein kinase B (PKB) is a member of the second messenger subfamily of protein kinases. The three isoforms of PKB identified have an amino-terminal pleckstrin homology domain, a central kinase domain, and a carboxy-terminal regulatory domain. PKB is the major downstream target of receptor tyrosine kinases that signal via the phosphoinositide (PI) 3-kinase. The crucial role of lipid second messengers in PKB activation has been dissected through the use of the PI 3 kinase-specific inhibitors wortmannin and LY294002. Receptor-activated PI 3 kinase synthesises the lipid second messenger PI-3,4,5-trisphosphate, leading to the recruitment of PKB to the membrane. Membrane attachment of PKB is mediated by its pleckstrin homology domain binding to PI-3,4,5-trisphosphate or PI-3,4 bisphosphate with high affinity. Activation of PKB alpha and beta is then achieved at the plasma membrane by phosphorylation of Thr308/309 in the A-loop of the kinase domain and Ser473/474 in the carboxy-terminal regulatory region, respectively. The upstream kinase that phosphorylates PKB on Thr308, termed PI dependent protein kinase-1, has been identified and extensively characterised. A candidate for the Ser473/474 kinase, termed the integrin-linked kinase, has been identified recently. Activated PKB is implicated in glucose metabolism, transcriptional control, and in the regulation of apoptosis in many different cell types. Stimulation of PKB activity protects cells from apoptosis by phosphorylation and inactivation of the pro-apoptotic protein BAD. These results could explain why PKB is overexpressed in some ovarian, breast, and pancreatic carcinomas. PMID- 10454218 TI - Antisense oligonucleotide inhibition of serine/threonine kinases: an innovative approach to cancer treatment. AB - The identification of genes that confer a growth advantage on neoplastic cells and the understanding of the genetic mechanism(s) responsible for their activation have made possible a direct genetic approach to cancer treatment using nucleic acid therapeutics. Moreover, the ability to block the expression of individual genes that promote carcinogenesis provides a powerful tool to explore the molecular basis of normal growth regulation, as well as the opportunity for therapeutic intervention. One technique for turning off a single activated gene is the use of antisense oligodeoxynucleotides and their analogs for inhibition of gene expression. The serine/threonine kinases are involved in mediating intracellular responses to external signals, such as growth factors, hormones, and neurotransmitters, and are involved in cell proliferation and oncogenesis. Described herein are recent studies supporting the potential use of oligonucleotides targeting these kinases as chemotherapeutic agents for cancer treatment. The serine/threonine kinases included here are protein kinase A, protein kinase C, and c-raf-1 kinase. PMID- 10454219 TI - Ochratoxin A-binding proteins in rat organs and plasma and in different cell lines of the kidney. AB - In order to detect cellular proteins which bind the mycotoxin ochratoxin A (OTA) we coupled OTA covalently to horseradish peroxidase (HRP). The peroxidase activity of the conjugate was used to detect these proteins in Western (ligand) blot analysis. Only signals caused by OTA binding to proteins were viewable. HRP alone detected no proteins and OTA-HRP binding could be inhibited by free OTA. Several proteins from the rat intestine, liver, spleen, and kidney were detected by OTA. Also rat plasma proteins bind OTA which confirms previous findings. In all renal cell lines investigated (MDCK-C11, OK, LLC-PK1, IHKE, and SKPT) there are several proteins which bind OTA. Comparison of the PonceauS stain on the nitrocellulose sheet with the signal obtained from OTA-HRP unveiled proteins with high specific OTA binding. Especially, proteins with molecular masses between 55 and 60 kDa, 40 and 45 kDa and 25 and 30 kDa showed OTA binding in all samples. OTA was partially displaced by aspartame and phenylalanine from some but not all proteins. Binding to cytosolic and organellar proteins was comparable in all investigated cell lines. In the OK cell organellar compartment a 62 kDa protein is preferentially detected by OTA-HRP although virtually no protein band is detectable. In conclusion we have found a method to clearly detect proteins which bind OTA. With this new method we proved that OTA has the potential to bind to several proteins yet specific binding differs dramatically. Thus, highly specific binding of OTA possibly makes certain proteins a preferential target of OTA toxicity. Furthermore, binding contributes to intracellular accumulation of OTA, thus leading to a prolonged half life in the mammalian body and emphasises the toxic potential of this fungal metabolite. PMID- 10454220 TI - Anthraquinone-induced cell injury: acute toxicity of carminomycin, epirubicin, idarubicin and mitoxantrone in isolated cardiomyocytes. AB - Acute toxic effects of the antineoplastic anthraquinones carminomycin, epirubicin, idarubicin and mitoxantrone were studied in primary cultures of cardiomyocytes, which were isolated from adult rats. Both time- and concentration dependent changes of cell structure and viability (trypan blue exclusion) following incubation of myocytes with subclinical, clinical and toxic concentrations of the anthraquinones were examined by light microscopy. The area under the decay curve of viable and rod-shaped myocytes was used to express cytotoxicity of the drugs. Mitoxantrone was found to reduce cell viability and number of rod-shaped cells to the greatest extent, followed by carminomycin, idarubicin and epirubicin. A significantly lower accumulation in cardiomyocytes was obtained with epirubicin and idarubicin compared with carminomycin. An inhibitory effect on oxygen consumption by the cells occurred already at 0.1 microM with epirubicin, whereas inhibition caused by other anthraquinones was less pronounced. Our data indicate a weak association of net accumulation and the toxicity parameter IC50 for carminomycin and idarubicin. In contrast to these results, a more significant correlation of cytotoxicity and anthraquinone lipophilicity was found, which suggests that the lipophilic character of a particular anthraquinone may be an important factor in drug-induced acute cardiotoxicity. PMID- 10454221 TI - Changes in hepatic nitrogen metabolism in isolated perfused liver during the development of thioacetamide-induced cirrhosis in rats. AB - Changes in hepatic nitrogen metabolism in isolated perfused liver were studied during the induction of experimental cirrhosis by thioacetamide in female Sprague Dawley rats. Cirrhosis of the micronodular type developed during 12-week administration of thioacetamide. Despite an increase in food consumption for 4 weeks after the end of administration, the physiological changes characteristic of cirrhosis were maintained. The rate of urea excretion per unit liver weight was significantly decreased compared with pair-fed control rats both during and after thioacetamide treatment. During 4 weeks of thioacetamide treatment, the rate of urea production in perfused liver from a combination of 0.25 mM NH4Cl and 1 mM glutamine decreased slightly, without a decrease in the maximum rate of urea production from 10 mM NH4Cl. In cirrhotic rats, the rate of urea production in perfused liver from NH4Cl and/or glutamine decreased, with a decrease in the maximum rate of urea production. The Km of ureagenesis for NH3 was unchanged in cirrhotic livers. During 4 weeks of thioacetamide treatment, glutamate dehydrogenase activity decreased, but the thioacetamide-induced cirrhotic state had no effect on glutamate dehydrogenase or glutaminase activity. Glutamine synthetase activity was decreased in rats treated with thioacetamide for 4 or 12 weeks. These results are consistent with the hypothesis that the capacity for urea production from NH3 and amino acids is decreased in the development of cirrhosis. PMID- 10454222 TI - In vitro inhibition of liver monooxygenases by beta-ionone, 1,8-cineole, (-) menthol and terpineol. AB - The present study was undertaken to investigate the inhibitory effects of beta ionone, (-)-menthol, 1,8-cineole and alpha-terpineol on liver microsomal enzymes involved in the biotransformation of xenobiotic substances. The effects of beta ionone and the foregoing monoterpenoid compounds on the activity of pentoxyresorufin-O-depentilase (PROD), a selective marker for CYP2B1, were determined in a pool of liver microsomes prepared from phenobarbital-treated rats. On the other hand, the inhibitory effects of these substances on the activities of ethoxyresorufin-O-deethylase (EROD), a marker for CYP1A1, and methoxyresorufin-O-demethylase (MROD), a marker for CYP1A2, were investigated in a pool of hepatic microsomes from beta-naphthoflavone-treated rats. Beta-ionone caused a concentration-related reduction of PROD activity with an IC50 value as low as 0.03 microM. The analysis of alterations produced by beta-ionone on PROD kinetic parameters (Lineweaver-Burk double-reciprocal plot) suggested that inhibition is non-competitive (Ki = 89.9 nM). Although being less potent than beta-ionone, 1,8-cineole (IC50 = 4.7 microM), (-)-menthol (IC50 = 10.6 microM) and terpineol (IC50 = 14.8 microM) also proved to be in vitro inhibitors of PROD reaction. Results also revealed that beta-ionone was a weak inhibitor of EROD (IC50 >100 microM) and MROD (IC50 >200 microM). Neither 1,8-cineole nor terpineol -tested in concentrations up to 150 microM--caused any decrease of EROD activity while (-)-menthol, at a concentration as high as 160 microM, produced only a slight reduction of the reaction rate. Terpineol (up to 150 microM) did not induce any reduction of MROD activity while 1,8-cineole (IC50 >300 microM) and ( )-menthol (IC50 >300 microM) caused only slight decreases of the reaction rate. The potent inhibitory effects on CYP2B1 suggest that beta-ionone, and the other monoterpenoids tested, may interfere with the metabolism of xenobiotics which are substrates for this isoenzyme. PMID- 10454223 TI - Investigation of some immunological functions in a group of asphalt workers exposed to polycyclic aromatic hydrocarbons. AB - To investigate the effects of polycyclic aromatic hydrocarbons (PAHs) on the human immune system, analyses were performed on T-cell subsets (CD4+, CD8+), B cells (CD19+), serum immunoglobulin levels (IgG, IgM, IgA) and white blood cell percentages of 16 male workers who were employed in road paving operations and compared to 12 unexposed male controls. PAH exposure was assessed using urinary 1 hydroxy-pyrene (1-OHP) levels and was found to be significantly higher in workers than in the controls. While the CD4+ cell percentage and the CD4+/CD8+ ratio were significantly higher in the PAH-exposed group, the percentages of CD8+ and CD19+ cells were unchanged. There was also a significant enhancement in serum IgG levels and the percentage of monocytes in the workers compared to the control group. These data suggest that chronic exposure to PAHs may affect some immune functions in humans. PMID- 10454224 TI - In vitro exposure effects of cyclosporin A and bis(tri-n-butyltin)oxide on lymphocyte proliferation, cytokine (receptor) mRNA expression, and cell surface marker expression in rat thymocytes and splenocytes. AB - Rat thymocytes and splenocytes were exposed in vitro to the model compounds Cyclosporin A (CsA), an immunosuppressive drug, and bis(tri-n-butyltin)oxide (TBTO), an immunotoxic environmental contaminant. The lymphocyte transformation test (LTT), cytokine (receptor) mRNA expression (RT-PCR and dot blot hybridisation), and flow cytometry were evaluated as assays for in vitro immunotoxicity, at dose levels that did not show effects on viability, this being the aim of the study. LTT and RT-PCR proved useful assays. Lymphocyte transformation was suppressed by both compounds, while IL-2 mRNA expression was suppressed by CsA but not by TBTO, and both compounds suppressed IL-2R mRNA expression in splenocytes but not in thymocytes. Furthermore, the data obtained suggest that antiproliferative effects may be more relevant than apoptosis induction for TBTO induced thymus atrophy. PMID- 10454225 TI - Continued medical education through reading and observation. PMID- 10454226 TI - Early brain proton magnetic resonance spectroscopy and neonatal neurology related to neurodevelopmental outcome at 1 year in term infants after presumed hypoxic ischaemic brain injury. AB - This study investigated the accuracy of prediction of neurodevelopmental outcome at 1 year using cerebral proton magnetic resonance spectroscopy (MRS) and structured neonatal neurological assessment in term infants after presumed hypoxic-ischaemic brain injury. Eighteen control infants and 28 infants with presumed hypoxic-ischaemic brain injury underwent proton MRS investigation. Studies were carried out as soon as possible after the cerebral insult, most within 48 hours. Infants had an early structured neurological assessment at a median of 19 hours (range 0 hours to 9 days) from the presumed hypoxic-ischaemic insult and a late assessment at a median of 7 days (range 3 to 25 days) during recovery. The maximum cerebral peak-area ratio lactate:N-acetylaspartate measured by proton MRS accurately predicted adverse outcome at 1 year with a specificity of 93% and positive predictive value of 92%. Neurological assessment had a tendency for false-positive predictions. However, both early and late neurological examination can be used as a reliable indicator for a favourable outcome at 1 year having negative predictive values of 100% and 91% respectively. PMID- 10454227 TI - Age-related walking in children with spina bifida. AB - The aim of this study was to examine children with myelomeningocele (spina bifida) who were investigated according to the level of neurosegmental lesion and to determine the number who walked and their average age at commencement/cessation of walking. The data have been collected since 1978 at the Royal Children's Hospital, Melbourne during annual evaluations of patients with spina bifida and high-level paralysis. After screening for established criteria and assigning to a neurosegmental lesion level, 173 children were examined for their ability to walk, with or without aids, in a community setting. Thirty-five children had lesions at the thoracic level, and seven walked at a mean age of 4 years 6 months. Three children with a lesion at the thoracic level ceased walking at a mean age of 7 years 6 months. Ten children had lesions at the high-lumbar level (L1/2), and five walked at a mean age of 5 years 2 months. Three children who had lesions at the high-lumbar level ceased walking at a mean age of 6 years 11 months. Fifteen children had lesions at the mid-lumbar level (L3) and nine walked at a mean age of 5 years. Three ceased walking at a mean age of 7 years. Forty-five children had lesions at the low-lumbar level (L4/5) and 38 walked at a mean age of 3 years 10 months. Five ceased walking at a mean age of 9 years 1 month. Sixty-eight children had lesions at the sacral level; all of whom walked at an average age of 2 years 2 months. None had ceased walking by the end of the study period. Delay in achieving ambulation can be expected in all children with spina bifida, including those with low neurosegmental level lesions. Furthermore, some children with high lesion level, given the opportunity, might be expected to ambulate effectively in the community but may cease walking after 3 to 4 years of this activity, which is earlier than previously recorded. PMID- 10454228 TI - Spinal cord ultrasonography in children with myelomeningocele. AB - Secondary tethering of the spinal cord in people with myelomeningocele (MMC) is almost universal but there is a relatively low incidence of the secondary tethered cord syndrome (S-TCS). In view of this, we wish to explore the notion that cord pulsation, as demonstrated by ultrasound of the spinal cord, can become an independent measure for increased tension of the spinal cord in patients with MMC. One-hundred and six patients with MMC underwent ultrasonographic examination of their back, a thorough physical examination, and a careful review of their clinical history. Of these patients, 53 had MRI of their back. Cord pulsation was graded and the statistical relation between pulsation and various clinical and MRI findings was analysed. Age and arm span were not correlated with grades of cord pulsation. The absence of cord pulsation correlated with the presence of symptoms but this relation may be explained by MRI findings of certain structural cord malformations and not by tethering per se. Assuming that cord pulsation is a reliable marker for the tension within the spinal cord, the results of our study suggest that overstretching of the cord is not the main cause for the development of the S-TCS. PMID- 10454229 TI - Cutaneomuscular reflex responses recorded from the lower limb in children and adolescents with cerebral palsy. AB - Cutaneomuscular reflex (CMR) responses were recorded from lower-limb and trunk muscles in 27 subjects with cerebral palsy (CP) (spastic, 21; athetoid, six) and in neurologically healthy (control) subjects, aged 3 to 15 years, while standing. In the 21 subjects with spastic CP, but not in the six subjects with athetoid CP, CMR responses were more widely distributed between ipsilateral lower-limb and trunk muscles compared with age-matched control children. CMR responses in older subjects with CP were similar to younger control subjects, lacking supraspinally mediated, long-latency components. Short-latency, spinally-mediated, excitatory CMR components were seen simultaneously in pairs of distal, antagonistic lower limb muscles in half of the subjects with spastic CP, but in none of the control children. In subjects with spastic-type CP, the abnormal reflex responses indicate disordered spinal and supraspinal inputs to motor neurones, although there was no convincing correlation between these responses and the severity of spasticity. PMID- 10454230 TI - Does abnormal branching of inputs to motor neurones explain abnormal muscle cocontraction in cerebral palsy? AB - The common synaptic drive shared between two groups of motor neurones synchronizes the timing of discharges between the motor-neurone groups. Recordings were made of motor-unit discharges during cocontraction of ipsilateral pairs of thumb muscles in eight subjects with cerebral palsy (CP) aged 4 to 13 years and eight neurologically healthy subjects aged 4 to 12 years, and in pairs of lower-limb muscles in 21 subjects with CP and 21 control subjects, both aged 3 to 15 years. Common synaptic drive, likely to be derived at least partly from activity in branched corticospinal-tract neurones, produced motor-unit synchronization between pairs of thumb muscles in control subjects but was absent in all subjects with CP. Motor unit synchronization was not found between lower limb antagonist muscles that cocontract abnormally in CP, nor was synchronization present in more widely separated muscle pairs. Therefore, abnormal patterns of muscle activation and more widespread muscle reflex responses do not result from an abnormal distribution of common synaptic drive in CP. PMID- 10454231 TI - Does academic achievement in children with epilepsy change over time? AB - A 4-year follow-up study of academic achievement in children aged between 11 and 17 years with epilepsy or asthma was carried out to identify differences between the two samples and to identify change in achievement over time. Differences based on sex and seizure severity also were explored. There were 98 subjects in the group with epilepsy and 96 subjects in the group with asthma. Academic achievement in five areas (Composite, Reading, Mathematics, Language, and Vocabulary) was measured using school-administered group test scores. To explore change over time in condition severity, each child was categorized as having a low or high condition severity at baseline (time I) and again 4 years later, resulting in four groups: low-low, low-high, high-low, and high-high. There were too few cases in the low-high group to be included in the analyses. Data were processed using analysis of covariance (ANCOVA), intraclass correlation coefficients, and paired t tests. At follow-up the children with epilepsy continued to perform significantly worse in all five achievement areas than the children with asthma. Children with either inactive or low-severity epilepsy had mean scores comparable to national norms; those with high seizure severity had mean scores ranging from 3 to 5 points below national norms. No changes were found in academic achievement over time for either sample, even among those whose conditions improved. Although boys with high-severity epilepsy continued to have the lowest achievement scores, there was no trend for them to decline in achievement over time. PMID- 10454232 TI - Sturge-Weber syndrome: cerebral haemodynamics during seizure activity. AB - The aim of this study was to examine the haemodynamic response to seizures in three infants with Sturge-Weber syndrome by measuring regional cerebral blood flow using transcranial Doppler sonography and 99mTc HMPAO SPECT. Time-locked video/digital EEG recording was carried out for ictal studies. MRI was performed in all subjects. SPECT showed hemispheric hypoperfusion interictally in all three patients and also ictally in one of the three; a small region of hyperperfusion was seen on the same ictal scan in the latter, ie. the patient with interictal and ictal hypoperfusion. In the two older children middle cerebral artery velocity (MCAV) was reduced by between 29 and 62% in the middle cerebral artery of the predominantly affected hemisphere compared with the contralateral side. During seizures, increases of 6 to 30% in MCAV were recorded for the clinically seizing hemisphere compared with 24 to 170% for the contralateral side in four of the seizures recorded. In one infant, MCAV fell bilaterally during a seizure that generalized (-18 and -43% in the predominantly affected and contralateral side respectively). Sequential recordings in one infant suggested that, with time, the haemodynamic response to seizures of the unaffected hemisphere may decrease. These findings suggest that the venous malformation in SWS is associated with an impairment of the cerebral haemodynamic response to seizure activity. PMID- 10454233 TI - Outcome of bilateral periventricular nodular heterotopia in monozygotic twins with megalencephaly. AB - Bilateral periventricular nodular heterotopia (BPNH) is a neuronal migration disorder, characterized by grey-matter cellular rests in the periventricular regions. Patients frequently present with seizures during childhood or later in life. Monozygotic twin sisters were found to have enlarged head circumferences (>95th centile) during a routine medical visit at 6 months of age. A brain CT scan revealed grey-matter cellular rests in the subependymal regions extending to the subcortical white matter. In some areas these cell aggregates had nodular margins, consistent with the diagnosis of BPNH. At 6 years of age they are cognitively and neurologically normal. They have not had any seizures and their heads continue to grow along the 95th centile. PMID- 10454234 TI - Myoclonic encephalopathy and diabetes mellitus in a boy. AB - We describe an 18-month-old boy with insulin-dependent diabetes mellitus who developed idiopathic myoclonic encephalopathy (dancing eye syndrome) at 26 months of age. The neurological symptomatology (multifocal myoclonus, opsoclonus, ataxia, behavioural disturbance) developed within 10 to 14 days after presentation. Biological, neuroradiological, and scintigraphic examination excluded CNS infectious diseases, intoxication, or tumours. At onset of diabetes mellitus, anti-glutamic-acid decarboxylase (GAD) antibodies were observed, and markedly increased in titre when myoclonic encephalopathy occurred. Corticosteroid treatment resulted in a decrease in anti-GAD autoantibody titres and the disappearance of neurological disturbances. As GAD is expressed both in pancreatic beta-cells and cerebellar Purkinje cells, it is possible that a common autoimmune disorder in this patient may account for both the diabetes and myoclonic encephalopathy. PMID- 10454235 TI - Melatonin treatment of sleep-wake cycle disorders in children and adolescents. PMID- 10454236 TI - Familial temporal-lobe epilepsy. PMID- 10454237 TI - Novel somatic mutations in the VHL gene in Swedish archived sporadic renal cell carcinomas. AB - Frequent loss-of-function somatic mutations of the VHL gene have been detected in sporadic renal cell carcinoma (RCC), indicating that inactivation of the VHL gene plays a critical role in RCC. In this study, we collected 35 archived Swedish sporadic RCCs identified from an epidemiological study on occupational exposure and kidney cancer to test how well stored pathological specimens could be retrieved and analyzed for VHL mutations. Thirty specimens were successfully analyzed with PCR-SSCP and sequencing. Aberrant SSCP bands were detected in 16 out of the 30 samples (53%). Sequencing analysis of the aberrant bands revealed seven deletions, one insertion, one base substitution on a splicing site, six missense mutations, one silent mutation and several base substitutions in the 5' non-coding region and intron 1. Most were novel somatic mutations that have not been reported in sporadic RCC. The mutations were found in three types of non papillary RCC cases, i.e. 14 clear cells, one granular chromophilic and one sarcomatoid RCC. Interesting multiple mutations were found in three cases (5, 3, 2 mutations, respectively). PMID- 10454238 TI - An in vivo rat model for visualizing glioma tumor cell invasion using stable persistent expression of the green fluorescent protein. AB - In our studies examining mechanisms of brain tumor cell migration/invasion into host tissue, the RT2 rat glioma cell line was stably transfected utilizing a green fluorescent protein (GFP) DNA construct. Stable transfected RT2 cells demonstrate high GFP expression at initial passages, maintain expression over 30 passages and have similar in vitro and in vivo growth patterns relative to controls. In rat brain tissue, individual tumor cells can be detected without any post-treatment. Using flow cytometry and cell sorting, we are able to retrieve and culture GFP positive cells from tumor core as well as adjacent tissue and contralateral hemisphere of tumor-bearing animals. PMID- 10454239 TI - Frequent multiplication of chromosome 1q in non-invasive and papillotubular carcinoma of the breast. AB - Carcinogenesis is considered to be a multistep process that may involve cumulative genetic alterations; one of these mechanisms, gain of chromosomal material, has the potential to activate tumor-promoting genes in breast carcinogenesis. Using 12 polymorphic microsatellite markers on the long arm of chromosome 1 (1q), we examined 130 sporadic breast carcinomas for abnormalities in the copy numbers of these loci in tumor cells using a differential PCR method. We also sought correlations between alterations on 1q and several clinicopathological parameters. At every locus examined, a 2-3-fold increase in copy number of an allele in tumor material was observed in one third of the tumors (46 of 130, 35%), indicating 'multiplication' of 1q. This multiplication involved the entire long arm in majority of those tumors (43 cases, 93%). The multiplication of 1q was observed more frequently in non-invasive ductal and papillotubular histological types than in solid-tubular and scirrhous types (13/25, 52% vs. 27/90, 30%) (P = 0.041). The predominant chromosomal alterations on 1q in breast carcinomas are found to be multiplications rather than losses. The multiplication represents polysomy of the entire region of 1q, and may confer a growth advantage during development and/or progression of non-invasive ductal and papillotubular histologic types of breast carcinomas. PMID- 10454240 TI - Topical and intratumoral photodynamic therapy with 5-aminolevulinic acid in a subcutaneous murine mammary adenocarcinoma. AB - One of the most promising substances used in photodynamic therapy (PDT) is 5 aminolevulinic acid (ALA), which induces endogenous synthesis and accumulation of porphyrins in malignant cells. In this paper we have shown that both topical and intratumoral administration of ALA in a subcutaneously implanted mammary carcinoma produced a significant synthesis of porphyrins and subsequent sensitization to laser light. Porphyrin accumulation was greater when ALA was administered intratumorally and tumour/normal skin porphyrin concentration ratios were higher compared with topical application. Irradiation was optimal between 2 and 3 h after topical application of 50 mg of a 20% ALA cream and 2-4 h after intratumoral administration of 30 mg ALA/cm3. The pattern of tumour response evaluated as the delay of tumour growth was similar following either route of drug administration. Applications of PDT were performed once, twice or three times in the study. The response to successive applications was constant for the same tumour, indicating that no resistance was acquired. Microscopic analysis showed both induction of foci of necrosis and haemorrhage, morphological features of apoptotic cells and total absence of cellular immune response. This paper reports on PDT with topical ALA in a subcutaneous carcinoma leading to tumour growth delay. These findings may have great relevance in the treatment of cutaneous metastasis of mammary carcinomas. PMID- 10454241 TI - Effects of human platelet-derived growth factor-AB on sarcoma growth in vitro and in vivo. AB - Platelet-derived growth factor (PDGF) has been proposed to play an important role in the growth of tumors. In order to study the effects of PDGF-AB on tumor growth in vivo, sarcoma-bearing mice were treated with PDGF-AB. The tumors, a malignant fibrous histiocytoma and an osteosarcoma, had functional PDGF receptors in vitro, as demonstrated by stimulation of PDGF-AB using a [3H]thymidine incorporation assay. Immunohistochemistry also revealed that both sarcoma xenografts expressed PDGF receptors. The tumor-bearing mice were given human PDGF-AB for 14 days, either continuously by an intraperitoneally placed mini-osmotic pump, or by daily injections. No effects on tumor growth in vivo were observed, as measured by tumor volume, autoradiography or cell cycle distribution. The histological appearance and ploidy of the tumors remained unaltered. The results indicate that, although the tumor cells are stimulated by PDGF-AB in vitro, the in vivo milieu or tumor growth pattern may render the tumors less susceptible to exogenously administered PDGF-AB in vivo. PMID- 10454242 TI - Induction and suppression of murine CYP-mediated biotransformation by dithianon: organ- and sex-related differences. AB - With the aim of evaluating the co-carcinogenic properties of dithianon, the regio and stereo-selective hydroxylation of testosterone was used as a multibiomarker of effect for cytochrome P450 (CYP) changes. CYP-catalysed reactions have been studied in liver, kidney and lung microsomes from male and female Swiss albino CD1 mice treated i.p. with single (3 or 6 mg/kg body wt.) or repeated (3 mg/kg body wt. daily for 3 days) doses of this fungicide. Induction or suppression was recorded under various situations in different organs and sexes. In liver, all testosterone hydroxylase (TH) activities were increased in the single treatment from 2.8- (6beta-, 16alpha- and 16beta-TH activities) to 16-fold (2beta-TH activity) in males at the lower dose. In contrast, activities were reduced from 33.3% (16beta- and 17-TH activities, lower dose) to 66.4% (16beta-TH activity, higher dose) in females. In kidney, a similar pattern of modulation was achieved: induction from 2.9- to 5-fold (6beta- and 2alpha-TH activities, higher and lower doses, respectively) in males; suppression from 47.4 to 50.2% (2alpha- and 2beta TH activities, either at lower or higher doses) in females. In lung, a significant induction ranging from 7.1- to 29.3-fold (16alpha- and 2alpha-TH activities, respectively, lower dose) in males, and up to a 7-fold increase (2beta-TH activity, higher dose) in females was obtained. After repeated treatment, hepatic 6beta-, 16beta-, 2alpha- and 2beta-TH activities were reduced up to approximately 60% in males, whereas no effect was seen in females. In extrahepatic tissues, a generalized increase of different THs was observed. The increase of 6beta-TH activity (CYP3A-linked), one of the most representative isoforms in humans, was sustained in liver and kidney by means of Western immunoblotting, using rabbit polyclonal antibodies anti CYP3A1/2. On the whole, a complex pattern of induction/suppression of CYP-dependent reactions was achieved depending on sex and tissue. The data are consistent with co-toxic, co carcinogenic and promoting potentials of this fungicide and provide information of interest in evaluating the risk associated with human exposure. PMID- 10454243 TI - Chemopreventive activity of very low dose dietary tannic acid administration in hepatoma bearing C3H male mice. AB - Tannins are plant polyphenols comprising a heterogeneous group of compounds. Tannic acid is a common tannin found in tea, coffee, immature fruits, etc. and it has also been used as a food additive. An increasing body of experimental evidence supports the hypothesis that tannins exert anticarcinogenic activity in chemically induced cancers in animal models. In the present study, tannic acid was administered in very low doses in the drinking water of C3H male mice divided into three groups (75 mg/l, 150 mg/l and 300 mg/l). These animals carry a genetic defect and show a high incidence of spontaneous liver tumors (> 50%) at an age older than 12 months. The results showed a decrease in the overall incidence of hepatic neoplasms (adenomas plus carcinomas): 53.3% of animals in the control group developed hepatic neoplasms versus 33.3% in the group given a low dose of tannic acid, 26.6% in the group given a medium dose and 13.3% in the high dosage group. The difference was more pronounced in the animals with carcinomas: 4.44% of mice who received tannic acid developed carcinomas versus 33.3% of those in the control group. Tannic acid administration did not affect the PCNA labeling index of normal hepatocytes. It is concluded that tannic acid dietary intake in low doses can exert a strong dose-dependent chemoprotective activity against spontaneous hepatic neoplasm development in C3H male mice, most probably through antipromoting mechanisms. PMID- 10454244 TI - Progestins suppress estrogen-induced expression of vascular endothelial growth factor (VEGF) subtypes in uterine endometrial cancer cells. AB - Vascular endothelial growth factor (VEGF) contributes to the early advancement of uterine endometrial cancers that conserve hormone dependency via angiogenic activity. This process prompted us to study sex steroidal suppression of VEGF expression in Ishikawa cells (a line of well-differentiated uterine endometrial cancer cells). Estrogen transiently induced VEGF subtype (VEGF165 and VEGF121) secretion from Ishikawa cells. Progestins (progesterone, medroxyprogesterone acetate (MPA) and 17 alpha-hydroxyprogesterone) suppressed the estrogen-induced events. In conclusion, progestins could suppress VEGF-related angiogenic potential, which contributes to tumor growth in the early stage of uterine endometrial cancers that conserve estrogen dependency. PMID- 10454245 TI - Levels of serum cathepsin L and several glycosidases in patients operated for colorectal cancer. AB - The activities of several glycosidases and cathepsin L were determined in the blood serum of a control group of ten healthy humans in comparison with a group (group I: 32 subjects) of preoperative colorectal cancer patients (1 week before surgical exeresis) and with another two groups: group II, comprising 18 operated subjects (1 week after surgery), and group III, of 15 operated subjects (4 months after surgery). All subjects were 48-88 years old. Both 'enzyme activity' and 'specific activity' determinations of serum beta-galactosidase, alpha-L fucosidase and cathepsin L revealed peculiar profiles that differed from one another. Control values differed from those of some stages of the pathological groups, but not of others. These values were compared also with the levels of total, lipid- and glycoprotein-associated serum sialic acid. The usefulness of some assays (especially cathepsin L activity measurement) in the follow-up of the health status of humans operated for colorectal cancer is discussed. PMID- 10454246 TI - Regulation of angiogenesis in human hepatomas: possible involvement of p53 inducible inhibitor of vascular endothelial cell proliferation. AB - Mechanisms regulating angiogenesis in human hepatomas were analyzed. Huh7 hepatoma cells transplanted into athymic mice formed highly vascularized reddish tumors with abundant microvessels, while angiogenesis by PLC/PRF/5 hepatoma cells was less remarkable. However, the production of angiogenesis stimulators such as VEGF and IL6 by Huh7 cells was much less than by PLC/PRF/5 cells. In addition, the production of angiogenesis inhibitor TSP-1 by Huh7 cells was greater than by PLC/PRF/5 cells. Therefore, the difference in angiogenesis between these two hepatomas was not explained by the production of these known angiogenesis regulators. On the other hand, PLC/PRF/5 cells but not Huh7 cells secreted an inhibitor of the proliferation of vascular endothelial cells, which was enhanced by p53-gene transfer. These results suggest that the production of this p53 inducible angiogenesis inhibitor is responsible, at least partly, for the regulation of angiogenesis in human hepatomas. PMID- 10454247 TI - Uptake of glucose analogues by colonic tumour cells during growth and after treatment with hydroxyurea. AB - SW620 cells were grown in tissue culture flasks to various cell densities producing populations of cells with a range of proliferative indices. The uptake of the two glucose analogues, deoxy-D-glucose (DG) and 3-O-methylglucose (OMG) was determined and found to be associated with S-phase fraction. The strong correlation between DG and OMG uptakes suggested that proliferation-related changes in transmembrane transport accounted for the association with S-phase fraction. Treatment of SW620 cells with the cell cycle inhibitor hydroxyurea was found to increase the uptake of DG and OMG in a time-dependent manner. PMID- 10454248 TI - Insulin resistance was connected with the alterations of substrate utilization in patients with cancer. AB - To elucidate the contribution of insulin resistance to substrate utilization, insulin sensitivity and substrate oxidation were examined in 19 cancer patients and five normal controls using the euglycemic hyperinsulinemic glucose clamp technique combined with indirect calorimetry. Glucose uptake and storage were significantly decreased in cancer patients compared with that of controls. In cancer patients, both glucose storage and oxidation were positively correlated with metabolized glucose as measured by the M value in cancer patients. Decrease in glucose uptake was mainly a reflection of decreased glucose storage rather than glucose oxidation. Inversely fat oxidation was negatively correlated with both the M value and glucose oxidation in cancer patients. In cancer patients with insulin resistance, decreased glucose uptake was closely associated with a rapid decrease in glucose storage, an increase in fat oxidation and a mild decrease in glucose oxidation, suggesting that insulin resistance was connected with the alterations of substrate utilization which may induce host depletion. PMID- 10454250 TI - Failure of phenethyl isothiocyanate to inhibit hamster tumorigenesis induced by N nitrosobis(2-oxopropyl)amine when given during the post-initiation phase. AB - The chemopreventive influence of phenethyl isothiocyanate (PEITC) during the post initiation stage was investigated in the N-nitrosobis(2-oxopropyl)amine (BOP) initiated hamster tumorigenesis model. A total of 120 female 5-week-old hamsters were divided into six groups. Animals in groups 1-3, each consisting of 30 hamsters, were injected twice, subcutaneously, with BOP 7 days apart to effect initiation. Starting 1 week after the second BOP injection, hamsters in groups 1 and 2 were fed diets supplemented with 6 micromol/g and 3 micromol/g of PEITC, respectively, for 51 weeks. Animals in group 3 received a basal diet as an initiation positive control. Animals in groups 4-6, each consisting of ten hamsters, were given 6 micromol/g or 3 micromol/g of PEITC alone, or were non treated, matched negative controls for groups 1-3. At the termination of experimental week 52, the incidences and multiplicities of neoplastic lesions in the target organs including the pancreas, lung, liver and kidney were found to be comparable among the BOP-treated groups. The values for pancreatic adenocarcinomas as well as dysplastic lesions tended to increase although without statistical significance. Taken together with our previous finding that PEITC dramatically inhibited the initiation phase of BOP-induced pancreatic and lung tumorigenesis in hamsters, it can be concluded that PEITC specifically exerts chemopreventive effects only when given concomitantly with the carcinogen. PMID- 10454249 TI - Hepatotoxicity and consequently increased cell proliferation are associated with flumequine hepatocarcinogenesis in mice. AB - It has been reported that flumequine (FLU) induces hepatic tumors in mice when given orally for 18 months. We investigated possible underlying mechanisms using a two-stage mouse hepatocarcinogenesis model. After initiation with a single intraperitoneal injection of 100 mg/kg body weight diethylnitrosamine (DEN) or saline, male CD-1 mice were given 4000 ppm FLU in the diet or 500 ppm phenobarbital (PB) in drinking water for 9, 19, 24 or 30 weeks. Toxicity, evidenced by centrilobular swollen and polar hepatocytes with fatty droplets, infiltration of inflammatory cells and increased numbers of mitosis in hepatocytes, was apparent in the livers of mice treated with FLU at all time points, but its severity declined towards the termination. FLU did not induce cytochrome P-450 enzymes such as 1A1, 2B1 and 3A2 as assessed immunohistochemically, while positive expression of 8-hydroxy-2'-deoxyguanosine (8-OHdG) was increased in hepatocytes of both DEN + FLU and FLU groups compared with the relevant controls. In animals given PB, eosinophilic swelling of hepatocytes was prominent, and the hepatocytes showed strongly positive reactions for CYP 1A1 and 3A2. Altered cell foci were induced in the livers of FLU-treated animals both with and without DEN initiation, especially the former, and their development paralleled the degree of hepatic toxicity. These results suggest that FLU hepatocarcinogenicity in mice is dependent on hepatotoxic damage and consequently increased cell proliferation. Oxidative damage to DNA may also be a crucial factor. PMID- 10454251 TI - Low-protein diet promotes sodium chloride-enhanced gastric carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine in Wistar rats. AB - Sodium chloride (NaCl) initiates and promotes experimental carcinogenesis in rats. We recently found that a high-protein diet attenuates NaCl-enhanced gastric carcinogenesis in Wistar rats. To investigate the effect of a purified low protein diet on NaCl-enhanced gastric carcinogenesis induced by N-methyl-N'-nitro N-nitrosoguanidine (MNNG) in Wistar rats, rats were fed a purified diet with an equalized caloric content containing 1% or 2% NaCl and 25% casein (normal-protein diet) or 10% casein (low-protein diet) after oral treatment with MNNG for 25 weeks. In week 52, neither 1% nor 2% NaCl had a significant effect on gastric carcinogenesis in rats fed a normal-protein diet. However, oral administration of 2%, but not 1%, NaCl significantly increased the incidence of gastric cancers in rats fed a low-protein diet. Oral administration of 2% NaCl also significantly increased the bromodeoxyuridine (BrdU)-labeling index and the ornithine decarboxylase (ODC) activity and decreased apoptosis of gastric cancers in rats fed a low-protein diet. However, 2% NaCl had no significant effect on these three parameters in rats fed a normal-protein diet. These findings indicate that a low protein diet enhances the effect of NaCl in gastric carcinogenesis and that this enhancement may be mediated by increased cell proliferation and reduced apoptosis of gastric cancers. PMID- 10454252 TI - p27 expression as a prognostic factor of breast cancer in Taiwan. AB - p27Kip1 is a member of the Cip/Kip family of cyclin-dependent kinase inhibitors. It binds to a variety of cyclin/CDK complexes, inhibits kinase activity, and blocks the cell cycle. Absent or reduced p27 expression has been shown to be a significant predictor of poor survival in breast, colorectal, prostate, non-small cell lung and esophagus carcinomas. An immunohistochemical assay was performed on 169 patients with primary breast cancers to evaluate the biologic significance of p27 expression. Decreased p27 expression was significantly associated with high grade (P = 0.00025), negative estrogen receptor (P = 0.00004), and negative progesterone receptor (P = 0.0038) breast cancers. Univariate analysis reveals that p27 expression inversely correlated significantly with overall survival (P = 0.0001). By multivariate analysis, p27 predicted the overall survival independently (P = 0.0096). Our study indicates that p27 expression is an independent prognostic marker of breast cancer in Taiwan. PMID- 10454253 TI - Prognostic value of DNA ploidy response to chemotherapy in human osteosarcomas. AB - We analyzed the DNA ploidy alterations after preoperative chemotherapy in 30 patients with non-metastatic osteosarcomas of the extremities. All of the patients received intensive chemotherapy with doxorubicin, cisplatin and methotrexate as well as wide tumor resection. DNA ploidy was determined by DNA cytofluorometry using isolated and smeared cells from biopsied and resected tumors after preoperative chemotherapy. The results showed that 12 diploid and nine non-diploid osteosarcomas did not change their ploidy pattern, but nine non diploid tumors changed to a diploid pattern with the disappearance of the aneuploid cells. The nine patients with altered ploidy tumors had a better histologic response to chemotherapy and a better prognosis than the patients with non-altered tumors especially diploid tumors (P = 0.0138). Therefore, we conclude that a decrease in aneuploid cells after chemotherapy is closely correlated with a good prognosis in half of the cases of aneuploid osteosarcoma. These results also suggest that aneuploid cells are more chemosensitive than diploid cells in human osteosarcomas. PMID- 10454254 TI - Inhibitory effect of microfibril wheat bran on azoxymethane-induced colon carcinogenesis in CF1 mice. AB - Microfibril wheat bran (MFW), a processed dietary fiber prepared by milling of coarse wheat bran (WB), is softer and has a more pleasant taste than WB. In this study, we examined the inhibitory effect of MFW on azoxymethane (AOM)-induced colon carcinogenesis in female CF1 mice and compared its effect with that of WB and cellulose (CL). The mice were fed a modified AIN 76 A diet supplemented with either MFW, WB, or CL at a final concentration of 20% (w/w). Six weekly s.c. injections of AOM (10 mg/kg body weight) were administered per mouse commencing 1 week after the start of the feeding period. Control mice were injected with saline only. Thirty-three weeks after the initial injection, the mice were sacrificed, examined for tumors, and the cecal contents were analyzed to determine the moisture content and the concentrations of short-chain fatty acids (SCFA). The average number of total tumors per mouse in the MFW (2.9 +/- 0.6, P = 0.017) and WB (5.3 +/- 1.3, P = 0.373) diet groups was lower than that of the CL diet group (7.5 +/- 1.3), though there was no significant difference in tumor incidence (94.7%, 90.0% and 94.7%, respectively) between the groups. More than 90% of the tumors in each group were adenocarcinomas. The incidence of adenoma and that of carcinoma in situ in the MFW diet group (5.3% and 0%, respectively) were also lower than those in the CL diet group (26.3 and 26.3%, respectively; P = 0.180 and P = 0.046, respectively). Analysis of the cecal contents revealed a significantly higher moisture content and significantly higher concentrations of SCFA, butyrate in particular, in the MFW and WB diet groups. The results of this study indicate that the source and texture of dietary fiber can influence tumor development in CF1 mice, and more specifically that MFW is a promising and useful dietary supplement with properties serving to protect against the development of colon cancer. PMID- 10454255 TI - Association of cyclin D1 expression in lung cancer and the smoking habits of patients. AB - This study examined whether or not cyclin D1 expression is associated with the smoking habits of patients with non-small cell lung carcinomas (NSCLC). Immunohistochemistry was used to analyze 181 NSCLC samples for the expression of cyclin D1. Expression of cyclin D1 protein was found in 130 out of 181 cases (72%). A significant relationship between cyclin D1 expression and stage or histological classification was not observed. The carcinomas of smokers expressed cyclin D1 in 77% of the cases while carcinomas of non-smokers expressed this protein only 57% of the time (P < 0.01, Fisher's exact test). The correlation between smoking and cyclin D1 expression was maintained when the analysis was limited to squamous cell lung carcinomas. However, no correlation was found between cyclin D1 expression and the smoking habits of patients with adenocarcinomas. This can be explained by the fact that the development of adenocarcinomas--in contrast to squamous cell lung carcinomas--is not closely related to tobacco smoke. PMID- 10454256 TI - Evidence that ferric nitrilotriacetate mediates oxidative stress by down regulating DT-diaphorase activity: implications for carcinogenesis. AB - Ferric nitrilotriacetate (Fe-NTA) is a known complete renal carcinogen as well as renal and hepatic tumor promoter, which acts by generating oxidative stress in the tissue. However, the mechanism by which it generates this stress is not fully understood. In this study, we show that Fe-NTA down-regulates hepatic and renal quinone reductase (QR) activity dose dependently. The maximum decrease in the activity of QR was observed at 12 h in the liver and 6 h in the kidney following Fe-NTA treatment. However, at all other time points studied, QR activity was reduced. In addition, a parallel increase in protein carbonyl content, a sensitive indicator of tissue oxidative stress was observed both in the liver and kidney. The pretreatment of animals with antioxidants, butylated hydroxyanisole and butylated hydroxytoluene, prevented the observed inhibition in the activity of QR and enhanced the formation of protein carbonyl in both organs. These studies suggest that Fe-NTA-mediated generation of oxidant free radicals down regulates QR activity which may be responsible, at least in part, for the observed renal and hepatic injury and carcinogenic properties of Fe-NTA. PMID- 10454257 TI - Anti-metastatic activity of curcumin and catechin. AB - The inhibitory effects of curcumin and catechin on lung metastasis induced by B16F-10 melanoma cells were studied in female C57BL/6 mice. Curcumin and catechin significantly (P < 0.001) inhibited lung tumour formation (89.3% and 82.2%, respectively) and significantly increased the life span (143.9% and 80.8%, respectively). Moreover, lung collagen hydroxyproline and serum sialic acid levels were found to be significantly (P < 0.001) lower in treated animals compared to the untreated controls. Curcumin and catechin treatment (10 microg/ml) significantly inhibited the invasion of B16F-10 melanoma cells across the collagen matrix of the Boyden chamber. Gelatin zymographic analysis of the trypsin-activated B16F-10 melanoma cells sonicate revealed that curcumin- and catechin-treated zymograms did not show any metalloproteinase activity. Curcumin and catechin treatment did not inhibit the motility of B16F-10 melanoma cells across a polycarbonate filter in vitro. These findings suggest that curcumin and catechin inhibit the invasion of B16F-10 melanoma cells by inhibition of metalloproteinases, thereby inhibiting lung metastasis. PMID- 10454258 TI - Ethnic susceptibility to lung cancer: differences in CYP2E1, CYP1A1 and GSTM1 genetic polymorphisms between French Caucasian and Chilean populations. AB - Many investigators have reported an association between genetic polymorphisms of cytochromes P-450 CYP2E1, CYP1A1 or glutathione S-transferase Mu (GSTM1) and susceptibility to lung cancer. However, pronounced interethnic variations have been described in the frequencies of these polymorphisms, especially between Asians and Caucasians. The present study was set up to establish CYP2E1 (c1, c2 and C, D), CYP1A1 (m1, m2 and Ile, Val) and GSTM1 (null) allelic frequencies in Chileans (n = 96) who are an admixture of Native Americans and Caucasians (Spaniards). The rare allele frequencies were found to be 0.15 (c2), 0.21 (C), 0.23 (m2), 0.32 (Val) and 0.21 ('null' genotype). These values are significantly higher than those of Caucasians except for the GSTM1 'null' genotype and suggest differences in susceptibility to lung cancer between both populations. PMID- 10454259 TI - Progression-linked overexpression of c-Met in prostatic intraepithelial neoplasia and latent as well as clinical prostate cancers. AB - The c-met proto-oncogene encoding the receptor for the hepatocyte growth factor is expressed in several cancers. In the present study, c-met protein (c-Met) was detected in eight of 22 (36%) cases of prostatic intraepithelial neoplasia (PIN), five of 15 (33%) latent and 17 of 21 (81%) clinical prostate cancers, including seven metastatic lesions, using an immunohistochemical method. All seven (100%) metastatic lesions investigated demonstrated strong staining, and a correlation between c-Met expression and histology was observed. These results suggest a significant relationship between c-Met expression and progression of prostate neoplasms, including latent cancers. PMID- 10454260 TI - Prognostic significance of tumor angiogenesis in primary fallopian tube cancer. AB - Tumor angiogenesis has been found to be prognostically significant in many types of malignant tumors. We assessed tumor vascularity in 43 cases of histologically proven primary fallopian tube cancer, FIGO stage I-IV, using the highly specific endothelial cell marker CD34. Microvessel count was determined by counting CD34 positive cells at 200 x magnification. The 5-year disease-free survival probability was 43.8% (+/- 11.5%) in 24 patients whose tumors had a microvessel count < or = 19 microvessels/field and 19.7% (+/- 9.5%) in the > 19 microvessels/field group (P = 0.046). Stage and microvessel count were statistically significant for disease-free survival in univariate analysis. Therefore, a larger sample size would be required to detect an independent and statistically significant prognostic effect of microvessel density in primary fallopian tube cancer in multivariate analysis. PMID- 10454261 TI - Telomerase enzyme activity and RNA expression in adriamycin-resistant human breast carcinoma MCF-7 cells. AB - Telomerase activity has been reported in cancer cells after treatment with antineoplastic agents. Assessment of telomerase activity could be a valuable tool to measure the reduction of aggression caused by chemotherapy. This study was designed to investigate the significance of telomerase for chemotherapy with respect to Adriamycin (ADM)-resistance. MCF-7 and its ADM-resistant line (AdrR) were treated with ADM, 5-fluorouracil (5FU) or taxotere (TAXO). Telomerase activity and human telomerase RNA component (hTR) were quantitatively measured by the telomeric repeat amplification protocol assay and RT-PCR, respectively. Cell counting and MTT assay were also performed. In MCF-7, enzyme activity was significantly reduced by ADM and 5FU treatments. In AdrR, 5FU and TAXO reduced enzyme activity, while ADM significantly increased the activity. No significant changes in hTR were seen in these two cell lines after treatment with any of these drugs. When Bcl-2 expression was examined after drug treatments, ADM increased Bcl-2 expression in AdrR cells, while not changing it in MCF-7 cells. We conclude that an unusual reaction of telomerase activity in AdrR may explain, at least in part, one of the mechanisms of the malignant biological behavior related with the drug-resistance to ADM. PMID- 10454262 TI - Distinct pattern of PCR-SSCP analysis of p53 mutations in human astrocytomas. AB - Clarification of somatic mutations during the progression of human astrocytomas is important in order to understand the mechanisms underlying the development of these tumors. We analyzed surgical specimens of human astrocytomas for mutations in the p53 gene using single-strand conformation polymorphism analysis of polymerase chain reaction product (PCR-SSCP analysis) at a low pH. Klenow fragment treatment after PCR amplification was an effective means to get rid of some extra bands on the SSCP gel. Five mutations in three of 24 astrocytomas were identified by this improved SSCP method. The frequency of p53 gene mutations in astrocytomas examined was 12.5%. Further examination by direct sequencing showed that all five mutants had single-base substitutions resulting in missense mutations. The present studies revealed a loss of heterozygosity and two point mutations on the remaining allele in one of the fibrillary astrocytomas. Finally, the improvement of PCR-SSCP analysis using Klenow treatment and low pH showed a distinct electrophoresis gel pattern and could be relevant for the prognosis of human astrocytomas. PMID- 10454263 TI - A new boronated porphyrin (STA-BX909) for neutron capture therapy: an in vitro survival assay and in vivo tissue uptake study. AB - A new boronated porphyrin compound (STA-BX909) was developed as a possible agent for boron neutron capture therapy. The boron concentration was measured by an in vivo rat experimental brain tumor model and an in vitro cell culture study. This agent was compared to sodium borocaptate (BSH) which has been used in clinical trials of boron neutron capture therapy. In the 9L rat brain tumor model, STA BX909 achieved a higher boron tumor/blood ratio 24 h after injection in comparison to BSH. A boron concentration study in cultured glioma cell lines (U 251, U-87, 9L) demonstrated an increased boron concentration as a function of exposure time to STA-BX909, while the boron concentration remained stable with increasing exposure time to BSH. Use of a colony forming assay with thermal neutron irradiation revealed more cytotoxicity with STA-BX909 than BSH when the same concentration of 10B was administered. We concluded that STA-BX909 may be an effective drug for use in boron neutron capture therapy and that it merits further investigation. PMID- 10454264 TI - Inhibition of liver glutathione S-transferase placental form-positive foci development in the rat hepatocarcinogenesis by Porphyra tenera (Asakusa-nori). AB - The effects of Asakusa-nori, Porphyra tenera (PT), a popular edible seaweed in Japan, on the development of putative preneoplastic lesions, glutathione S transferase placental form (GST-P)-positive foci, in the male F344 rat liver were examined using a medium-term bioassay system. PT significantly decreased both the number and area of GST-P-positive foci in rat livers initiated with diethylnitrosamine (DEN). To investigate possible mechanisms of inhibition, effects of PT on 5-bromo-2'-deoxyuridine (BrdU) labeling in GST-P-positive foci and the surrounding area of hepatocytes were studied. The ratio of the GST-P positive foci to surrounding tissue labeling indices was decreased in the PT treated group as compared with the DEN alone group. Ornithine decarboxylase activity in the liver was slightly increased and spermidine/spermine N' acetyltransferase activity was slightly decreased in the PT-treated animals. These results suggest that PT possesses chemopreventive effects against DEN induced hepatocarcinogenesis. PMID- 10454265 TI - Transplacental administration of diethylstilbestrol (DES) causes lesions in female reproductive organs of Donryu rats, including endometrial neoplasia. AB - The effects of transplacental administration of diethylstilbestrol (DES) on female reproductive organs were investigated using Donryu rats. The animals were given subcutaneous injections of DES dissolved in olive oil at doses of 0.01 or 0.1 mg/kg on days 17 and 19 of gestation. In female offspring, clinical signs, body weights and estrous cycles were continuously assessed until all survivors were killed at month 18. A low mean litter size and shortening of period of pregnancy were recognized in the 0.1 mg/kg group. Disorder and/or suspension of the estrous cycle (so called persistent estrus) also appeared very early in the 0.1 mg/kg group. Macroscopically, the incidences of hypoplasia of the oviduct, cystic dilatation of the uterus and small size of the uterine cervix were higher in the 0.1 mg/kg group than those in the control group. Histologically, in the ovary, the incidence and degree of atrophy were increased in both 0.01 and 0.1 mg/kg groups. In the uterus, total incidences of endometrial hyperplasias were about the same in all groups. However, endometrial adenocarcinomas were dose dependently increased in the treated groups, the incidence in the 0.1 mg/kg group being significant, compared to that in the control. In the vagina, mucification was more prominent in the treated animals, especially at the higher dose, but no tumors were observed. The present results indicate that prenatal exposure to DES can produce uterine adenocarcinomas in rats, as reported earlier for mice, although its carcinogenic activity is not so strong. Increase of endometrial adenocarcinoma incidence might depend on hormonal imbalance resulting from the ovarian atrophy due to transplacental treatment of DES. PMID- 10454266 TI - The role of the coil click in TMS assessed with simultaneous EEG. AB - OBJECTIVE: We have used EEG to measure effects of air- and bone-conducted sound from the coil in transcranial magnetic stimulation (TMS). METHODS: Auditory evoked potentials to TMS were recorded in three different experimental conditions: (1) the coil 2 cm above the head, (2) the coil 2 cm above the head but rigidly connected by a plastic piece to the scalp, (3) the coil pressed against the scalp over the motor cortex. RESULTS: The acoustical click from the TMS coil evoked large auditory potentials, whose amplitude depended critically on the mechanical contact of the coil with the head. CONCLUSION: Both air- and bone conducted sounds have to be taken into account in the design and interpretation of TMS experiments. PMID- 10454267 TI - A method for generating natural-sounding speech stimuli for cognitive brain research. AB - OBJECTIVE: In response to the rapidly increasing interest in using human voice in cognitive brain research, a new method, semisynthetic speech generation (SSG), is presented for generation of speech stimuli. METHODS: The method synthesizes speech stimuli as a combination of purely artificial processes and processes that originate from the natural human speech production mechanism. SSG first estimates the source of speech, the glottal flow, from a natural utterance using an inverse filtering technique. The glottal flow obtained is then used as an excitation to an artificial digital filter that models the formant structure of speech. RESULTS: SSG is superior to commercial voice synthesizers because it yields speech stimuli of a highly natural quality due to the contribution of the man originating glottal excitation. CONCLUSION: The artificial modelling of the vocal tract enables one to adjust the formant frequencies of the stimuli as desired, thus making SSG suitable for cognitive experiments using speech sounds as stimuli. PMID- 10454268 TI - Spectral versus visual EEG analysis in mild hepatic encephalopathy. AB - OBJECTIVE: Spectral EEG analysis has been claimed to reduce subjective variability in EEG assessment of hepatic encephalopathy and to allow the detection of mild encephalopathy. METHOD: To test such assumptions, 43 digital EEG were recorded in 32 cirrhotics without overt encephalopathy or with grade 1 overt encephalopathy; 7 patients were re-tested (2-5 times) in their follow up. All patients underwent psychometric assessment. Nineteen controls were considered. EEG were blindly evaluated by two electroencephalographers and by spectral EEG analysis performed according to 3 different techniques. RESULTS: The reliability of the classification based on spectral analysis (biparietal technique) was higher than that based on a three-degree qualitative visual reading (concordance/discordance = 58/4 versus 46/16 P < 0.01) and comparable with that of semiquantitative visual assessment based on posterior basic rhythm (concordance/discordance = 55/7 P = 0.5). The accuracy of spectral EEG analysis was higher than that of qualitative visual EEG readings alone (90 versus 75%) and comparable to semi-quantitative visual assessment (87%), however, statistical significance was not reached. In the follow-up, the variations of theta and delta relative power were found to be significantly correlated with psychometric variations. CONCLUSIONS: In conclusion, spectral EEG analysis may improve the assessment of mild hepatic encephalopathy by decreasing inter-operator variability and providing reliable parameters correlated with mental status. PMID- 10454269 TI - P300 to high intensity stimuli during REM sleep. AB - OBJECTIVES: The P300 component of the auditory event-related potential (ERP) is a large amplitude positive wave peaking at approximately 300 ms following detection of a rare significant stimulus. Although P300 is typically only evoked when the subject attends to the stimulus, it may also be elicited in an awake, inattentive subject if the stimulus is sufficiently intrusive. We therefore employed an oddball task to determine if high intensity stimuli would elicit the P300 during sleep. METHODS: A loud 90 dB SPL tone pip was presented infrequently (P = 0.05) in a train of lower intensity 70 dB SPL standard stimuli. A multiple channel EEG was recorded from 8 good sleepers during a single night. RESULTS: A large amplitude parieto-central P300, peaking at 321 ms, was apparent in REM sleep to the loud deviant stimulus. In stage two non-REM sleep, a later positive wave, peaking at 446 ms, was apparent even after K-Complexes were removed from the average. This non-REM P450 was however maximum over occipito-parietal areas of the scalp. CONCLUSION: The presence of a P300 in REM sleep following a loud, rare stimulus indicates that sensory discrimination capabilities remain intact during this state. This may be associated with either pre- or conscious processing of relevant stimuli. PMID- 10454270 TI - Spatial structure of the human alpha rhythm: global correlation in adults and local correlation in children. AB - OBJECTIVE: The maturation of the neocortex during childhood and adolescence involves dramatic increases in white-matter volume. EEG recordings from children and adults were examined to determine whether there are associated changes in spatial properties of dynamic processes in the neocortex. METHODS: Spontaneous eyes-closed and eyes-open EEG were recorded at 128 electrodes in 20 children aged 6-11 years and 23 adults aged 18-23 years. The surface Laplacian algorithm was applied to improve the spatial resolution of each electrode. Power and coherence were used to characterize the spatial structure of the alpha rhythm. A stochastic field model was used to eliminate coherences that are inflated due to volume conduction. RESULTS: In adults, the alpha rhythm is characterized by very high coherence between distant electrodes. The children demonstrated reduced anterior power and coherence between anterior and posterior electrodes at the peak alpha frequency in comparison to the adults. The Laplacian alpha rhythm demonstrated much higher power in the children at both anterior and posterior electrodes, but was weakly correlated between electrodes. CONCLUSIONS: The maturation of neocortex in late childhood involves increased global correlation by long-range corticocortical connections. The local correlation that contributes power to each electrode, independent of other electrodes, is reduced as the global correlation increases. PMID- 10454271 TI - Early neurological and neuropsychological development of the preterm infant and polyunsaturated fatty acids supply. AB - OBJECTIVES: To compare the early neurological maturation of premature newborns (PT) fed breast milk (BM) or a formula containing only 18-carbon polyunsaturated fatty acids (PUFA) (A) or enriched with long chain (LC) PUFA (B). METHODS: PT enrolled the 2nd day of enteral feeding (D0) were fed BM (n = 15; 4 dropped out) or randomly assigned to A (n = 11; 2) or B (n = 14; 1) for at least 30 days (D30). Auditory and visual evoked potentials (EPs) and nerve conduction velocity (NCV) and plasma and red blood cell (RBC) phospholipid composition were determined at D0 and D30. No difference was found between groups for the D0-D30 changes in EP parameters. The maturation of motor NCV was slower in the B group than in the two other groups. In plasma, the changes were higher in B than in the BM and A groups for linoleic acid (P < 0.05), in BM versus B group for arachidonic acid (P < 0.02). In RBC, formula groups displayed higher linoleic acid level than the BM group (P < 0.05). No difference was found between groups for the changes in arachidonic and docosahexaenoic acids. CONCLUSIONS: A balanced supply of n-6 and n-3 PUFA without addition of LC-PUFA allowed an adequate early maturation of the central nerve system. The effects of LC-PUFA on the maturation of NCV remain to be confirmed. PMID- 10454272 TI - Cortical activation related to auditory semantic matching of concrete versus abstract words. AB - OBJECTIVE: The aim of this study was to examine cortical correlates of semantic memory processes in the auditory stimulus modality. METHODS: Event-related desynchronization (ERD) and synchronization (ERS) of the lower (8-10 Hz) and upper (10-12 Hz) alpha frequency bands of background EEG were studied in 10 subjects performing an auditory semantic matching task. The stimuli were abstract and concrete nouns which were presented sequentially in pairs. The task was to decide whether the two nouns belonged to the same semantic category or not. RESULTS: The presentation of the first stimulus (encoding) elicited ERS whereas the presentation of the second stimulus (semantic matching) elicited ERD. Abstract nouns presented as the second stimulus elicited ERD which was most prominent in the lower alpha frequency band, whereas the presentation of a concrete noun as the first stimulus elicited ERS, most prominently in the upper alpha frequency band. CONCLUSIONS: The auditorily elicited ERD/ERS reflects cortical activity associated with cognitive functions. The present findings demonstrate that the auditorily elicited ERD/ERS can reveal subtle differences in auditory information processing. Semantic memory processes (encoding and comparison) are reflected as varying responses in the two alpha frequency bands. PMID- 10454273 TI - Early occipito-parietal activity in a word recognition task: an EEG and MEG study. AB - OBJECTIVE: We investigated early brain activity (under 200 ms after the stimulus onset) related to the encoding and the retrieval of verbal information. METHODS: First, we compared ERPs produced by words which were encoded to ERPs produced by words from following test phases (correctly identified repetitions and correctly classified new words) in two different experiments. Experiment 1 consisted of an intentional learning paradigm and experiment 2 consisted of an incidental learning paradigm. In addition, we conducted a control experiment (experiment 3), which was a continuous recognition task with two different repetition intervals. Secondly, we conducted a magnetoencephalographic (MEG) study to further investigate early brain activity (experiment 4). The same intentional learning paradigm as in experiment 1 was used. RESULTS: We found that ERPs elicited by correctly classified test words (repeated words and new words) of both experiment 1 and experiment 2 were significantly more negative going than the ERPs elicited by the study words. This effect was apparent between 100 ms and 200 ms after the stimulus onset and was distributed over occipital and parietal scalp locations. In the control task (experiment 3), these early potential differences were missing (for both repetition intervals). Early event-related fields (ERFs) were also found to depend on the situation of the study phase and the test phase. This activity difference peaked at 120 ms after the stimulus onset. The distributions of the difference magnetic fields were occipito-parietal and thus consistent with the findings of experiment 1 (EEG-experiment). CONCLUSION: Whether the effect we defined in the present study is due to an increase of activity during the test situation or due to a decrease of activity during the study situation remains unclear. However, it might reflect attentional processes within a word recognition task depending on whether a word is encoded or an effort of word retrieval has to be made. PMID- 10454274 TI - Test-retest reliability of mismatch negativity for duration, frequency and intensity changes. AB - OBJECTIVES: The test-retest reliabilities of the mismatch negativity (MMN) elicited by deviances in sound duration, frequency, and intensity were compared. METHODS: The MMN was recorded with a 64-channel electroencephalograph (EEG) from 15 healthy adult subjects in two sessions over intervals of 1-27 days. During the recordings, subjects watched a silent movie while they were presented with one long stimulus sequence consisting of 6 types of tones. The standard tone (P = 0.8) of 75 ms in duration consisted of 3 lowest harmonic partials with 500 Hz as the fundamental frequency. Each of the 5 different deviant tones was presented with P = 0.04: frequency deviants (+/-5%, +/-10%), duration deviants (-66%, 33%), and intensity deviants (- 15 dB). RESULTS: The 66% duration decrement elicited MMN with the most replicable amplitude (r = 0.78) and latency (r = 0.76) among the deviances tested. CONCLUSIONS: Since these reliabilities considerably exceed those reported previously, these data support the use of the duration decrement deviance presented with spectrally rich tones while investigating the integrity of the patients' cognitive brain functions using the MMN. PMID- 10454275 TI - Shortening of the cortical silent period following transcranial magnetic brain stimulation during an experimental paradigm for generating contingent negative variation (CNV). AB - OBJECTIVES: We investigated changes in the cortical silent period (CSP) following transcranial magnetic cortical stimulation (TCMS) during a standard paradigm which was designed to evoke contingent negative variation (CNV) in ten normal subjects. METHODS: We recorded the motor evoked potentials (MEP) and CSP during the inter-stimulus interval (ISI) of a CNV paradigm in ten normal subjects. The CNV paradigm consisted of a visual warning stimulus (S1) followed by a visual response stimulus (S2). The CSP following TCMS on the hand motor area was recorded from mildly contracted first dorsal interosseous muscles. RESULTS: The CSP was significantly shortened during the ISI (P < 0.01, t test) with a highly significant correlation with the TCMS timing during the ISI (P < 0.02, Spearman's correlation coefficient), while the MEP amplitude and latency were unchanged. CONCLUSIONS: The results suggested that shortening of the CSP was associated with neural processes related to preparation for voluntary movement during the paradigm. PMID- 10454276 TI - Life-span changes in EEG spectral amplitude, amplitude variability and mean frequency. AB - OBJECTIVES: The objectives were to assess life-span alterations in intrasubject variability of EEG spectral amplitudes and in amplitude and mean frequency computed from spectral amplitude profiles. METHODS: EEG was recorded from the central and occipital scalp of 222 healthy males aged 4 to 90 years. Amplitude spectral profiles derived from FFT procedures provided data for the computation of amplitude variability (ampCV), absolute and relative amplitude and mean frequency for each of five spectral bands. Analysis of variance, coefficient of correlation and t test were employed in data analyses. RESULTS: The ampCV measure did not provide direct evidence of age-related intrasubject changes in EEG frequency but was useful for indexing the presence of rhythmic EEG activity. Marked decreases in absolute amplitude occurred during childhood with little change thereafter. Age changes in relative amplitude were more complex and differed for slow and fast EEG activity. We also report that an algorithm frequently used to estimate mean frequency can introduce consistent bias into mean frequency computations under some conditions. CONCLUSIONS: Life-span changes in measured EEG characteristics were generally consistent with those from earlier studies. It is important that investigators validate and describe procedures used to determine mean frequency of EEG spectral data. PMID- 10454277 TI - Instantaneous EEG coherence analysis during the Stroop task. AB - OBJECTIVE: In the present study the Stroop effect is analyzed by means of EEG coherence analysis in addition to traditional analysis of behavioral data (reaction time) and ERP analysis. Data from 10 normal subjects are examined. METHODS: In particular, a special dynamic approach for a continuous coherence estimation is applied to investigate the procedural evolution of functional cortical relationships during the Stroop task. RESULTS: The frequency band of 13 20 Hz is found to be sensitive to the discrimination between the congruent and the incongruent task conditions on the basis of instantaneous coherence analysis. The magnitude of coherence values within the time interval of late potentials and the maximal coherence values are used to assess the strength of interaction between distinct areas of the cortex. Higher coherences are observed within the left frontal and left parietal areas, as well as between them for the incongruent situation in comparison with the congruent situation. Furthermore, the time points of maximal coherence allows a procedural discrimination between both situations. The peak synchrony described by the time-points of maximal coherence correlates strongly with the reaction times mainly within the frontal area and between fronto-parietal areas in the incongruent case, whereas this correlation is restricted to the right hemisphere in the congruent case. PMID- 10454278 TI - The technical, neurological and psychological significance of 'alpha', 'delta' and 'theta' waves confounded in EEG evoked potentials: a study of peak latencies. AB - It is suggested that lack of progress in understanding the neurological and psychological significance of EEG averaged evoked potentials (AEPs) and event related potentials (ERPs) may be due to the confounding of, 'delta', 'theta' and 'alpha' frequency responses generated by the 'brain-stem' 'limbic' and 'thalamic' cortical activating systems, respectively. This hypothesis was tested in a sample of 93 adults by recording EEG responses to auditory tones and using narrow-band filters to distinguish '4 Hz', '7 Hz', and '10 Hz' component waves in the unfiltered AEP. Bivariate and multivariate statistical analyses of peak latencies provided evidence of the distinct and unitary nature of these component waves and demonstrated their relative influence on the P1, N1 and P2 peaks of the unfiltered AEP. The results also reveal a predicted negative relationship between the 10 and 4 Hz latencies which is attributed to inhibition of the delta frequency brain-stem ascending reticular activating system (BSARAS) by the alpha frequency diffuse thalamic projection system (DTPS). Correlations of the 4 Hz latencies, with age, sex, and behavioural arousal variables confirm that the 4 Hz latencies are associated with the maintenance of behavioural arousal, an important function of the BSARAS. PMID- 10454279 TI - The electrical and magnetical cerebral responses evoked by electrical stimulation of the esophagus and the location of their cerebral sources. AB - OBJECTIVES: After electrical stimulation of the esophagus cerebral responses are recordable, their cortical source is under discussion. Brain mapping using electroencephalography recordings demonstrated partially controversial results. Sources of evoked responses can be localized more easily using magnetoencephalography than electroencephalography. METHODS: We examined 22 volunteers by recording electrical somatosensory potentials after electrical stimulation of the esophagus. In 9 of these 22 subjects additional recording of magnetic fields was performed and the sources of the evoked magnetic fields were computed. RESULTS: The evoked potentials after electrical stimulation of the esophagus had a similar latency as the previously published data. The source localization done by magnetoencephalography suggest that first a region of the postcentral gyrus is activated which is temporo-lateral to the primary somatosensory cortex of the pharynx. This region is suggested to be the primary somatosensory region of the esophagus. This source was followed by a source in the parietal operculum thought being part of the secondary somatosensory cortex. Simultaneously the insular cortex was activated pointing to a parallel neuronal pathway to the central autonomic nervous system. CONCLUSION: After electrical stimulation of the esophagus somatosensory cortical areas of the temporal postcentral gyrus and the operculum are activated. In parallel activation of the insular cortex as part of the central autonomic network was found. PMID- 10454280 TI - Reciprocal modulation of somatosensory evoked N20m primary response and high frequency oscillations by interference stimulation. AB - OBJECTIVES: We examined whether the inverse relation between somatic evoked N20m primary response and high-frequency oscillations during a wake-sleep cycle (Hashimoto, I., Mashiko, T., Imada, T., Somatic evoked high-frequency magnetic oscillations reflect activity of inhibitory interneurons in the human somatosensory cortex, Electroenceph clin Neurophysiol 1996;100:189-203) holds for interference stimulation. METHODS: Somatosensory evoked fields (SEFs) from 14 subjects were measured following electric median nerve stimulation at the wrist with, and without, concurrent brushing of the palm and fingers. SEFs were recorded with a wide bandpass (0.1-1200 Hz) and then N20m and high-frequency oscillations were separated by subsequent low-pass (< 300 Hz) and high-pass (> 300 Hz) filtering. RESULTS: The N20m decreased dramatically in amplitude during interference stimulation. In contrast, the high-frequency oscillations moderately increased in number of peaks. CONCLUSIONS: These results demonstrate the presence of an inverse relation between N20m and high-frequency oscillations for interference stimulation. We speculate that the high-frequency oscillations represent a localized activity of GABAergic inhibitory interneurons of layer 4, characterized by a high-frequency spike burst (200-1000 Hz) without adaptation, and that the continuous interference stimulation induces tonic excitation of the interneurons, leading to a facilitation of responses to the coherent afferent volley elicited by the median nerve stimulation (bottom-up mechanism). On the other hand, refractoriness of the pyramidal neurons caused directly by interference stimulation along with an enhanced feed-forward inhibition from the interneurons will lead to a decrease of N20m amplitude. PMID- 10454281 TI - The value of combined ambulatory cassette-EEG and video monitoring in the differential diagnosis of intractable seizures. AB - OBJECTIVE: To evaluate the ability of combined ambulatory cassette-EEG and video monitoring (ACV) to establish a diagnosis in patients with attacks of unknown nature and its impact on their treatment. METHODS: We evaluated ACV in 125 consecutive patients with attacks of unknown nature. Most had intractable attacks suspected of being non-epileptic. Antiepileptic drugs (AEDs) were discontinued or reduced at the start of the procedure. The median duration of monitoring was 3 days (range 1-10). The ACV results and patient records were reviewed, and patients were called for additional follow-up when feasible. RESULTS: Attacks were recorded in 101 patients. They were epileptic in 20 patients, psychogenic in 60, both in 3, and of unknown nature in 18 (usually subjective episodes). The study resulted in AED discontinuation at discharge in 46 patients with recorded psychogenic seizures and 6 with recorded attacks of unknown nature. Three quarters of patients followed up were free of attacks or improved. CONCLUSIONS: ACV was effective in providing a diagnosis in two-thirds of patients. If psychogenic seizures are suspected in patients on AEDs, ACV may provide the diagnosis and may help exclude epilepsy, without the need for standard EEG-CCTV, which can then be reserved for patients undergoing presurgical evaluation. PMID- 10454282 TI - Distribution of brainstem somatosensory evoked potentials following upper and lower limb stimulation. AB - OBJECTIVE: Because somatosensory evoked potentials (SEP) to lower limb stimulation have not been recorded from the brainstem to the extent that upper limb SEPs have been studied, we compared brainstem recordings in response to both median nerve (MN) and posterior tibial nerve (PTN)stimulation. METHODS: SEPs were recorded directly from the dorsal surface of the brainstem in four patients with fourth ventricle tumors. RESULTS: Following MN stimulation, medullary SEPs were characterized by a major negativity (N1) preceded by a small positivity (P1) and followed by a large positivity (P2). In the pons, triphasic waves with predominant negativity were obtained. With PTN stimulation, similar medullary SEPs with a P1'-N1'-P2' configuration and pontine SEPs with a triphasic waveform were obtained. CONCLUSIONS: Since the distribution of PTN SEP was identical to that of MN SEP, PTN SEPs are thought to be generated by mechanisms similar to those for MN SEP. Thus, the P1' and N1' of medullary SEP would be generated by the dorsal column fibers that terminate in the nucleus, with P2' possibly arising postsynaptically in the nucleus. The triphasic PTN SEP from the pons reflects an axonal potential generated in the medial lemniscal pathway. PMID- 10454283 TI - Median and ulnar palm-wrist studies. AB - OBJECTIVES: Routine carpal tunnel electrodiagnosis frequently includes median (MPW) and ulnar (UPW) palm-wrist mixed nerve conduction latency determinations over 8 cm. Despite widespread use, normative palmar latency difference (PLD) and UPW values, and the relative utility of onset latency (OL) or peak latency (PL) measurements are controversial. The current study was conducted to determine normative values for these parameters. METHODS: MPW and UPW studies were performed unilaterally in 33 normal controls. The PLD-OL and PLD-PL were calculated. The mean, range, standard deviation, and upper limits of normal were determined. 74 hands (50 patients) with both clinical and electrophysiologic median neuropathy were also studied. RESULTS: The abnormal MPW and UPW cut-offs were both 1.8 ms (OL), and 2.3 ms (PL). The abnormal PLD cut-offs were 0.5 ms (OL and PL). Using either OL or PL, PLD parameters were similar within controls, and also within CTS patients. Using either OL or PL, UPW parameters were similar between controls and CTS patients. CONCLUSIONS: An abnormal PLD cut-off of 0.5 is recommended. This is slightly higher than some prior recommendations, however it should minimize the likelihood of false positive studies. Onset and peak latency measurements are likely to have similar clinical utility. PMID- 10454284 TI - Macro-EMG in mitochondriopathy. AB - OBJECTIVES: In a previous macro-EMG study on patients with mitochondriopathy, macro-MUAP amplitude has been shown to be the electromyographic parameter most often abnormal. We wanted to investigate the role of the macro-MUAP area and if macro-EMG is more helpful than conventional needle-EMG in detecting involvement of the skeletal muscle in patients with mitochondriopathy. METHODS: From the right brachial biceps muscles of 38 healthy subjects and 20 patients with primary mitochondriopathy, aged 23-72 y, conventional needle EMGs and macro-EMGs were recorded one after the other. RESULTS: Macro-MUAP amplitude and macro-MUAP area were not significantly different between patients and controls. The sensitivity of the macro-EMG was 35%, that of conventional needle EMG 40%. Both methods supplemented each other. CONCLUSIONS: The sensitivity of the macro-EMG to detect involvement of the skeletal muscle in patients with mitochondriopathy is similar to that of conventional needle EMG. Macro-MUAP area was not more helpful than macro-MUAP amplitude in this respect. PMID- 10454285 TI - Exclusive electrophysiological motor involvement in carpal tunnel syndrome. AB - OBJECTIVE: To define the frequency of exclusive electrophysiological motor involvement in carpal tunnel syndrome (CTS). METHODS: We reviewed the electrophysiological studies of 2727 consecutive hands with typical symptoms and signs of CTS and at least one abnormal test of the following: median distal motor latency (DML), digit two sensory conduction velocity (D2-SCV), segmental D2-SCV from wrist to palm, median-ulnar sensory latency difference from ring finger stimulation. RESULTS: Thirty-one hands (1.2%) had prolonged median DML ( > 4.4 ms) with normal SCV ( > 48 m/s). In 17 of 31 hands, segmental D2-SCV from wrist to palm or median-ulnar latency difference from ring finger stimulation were also performed with normal results in 8 hands, demonstrating a true exclusive electrophysiological motor involvement. CONCLUSIONS: In CTS, exclusive electrophysiological involvement of median motor fibers is rare. It may be related to preferential compression of the intraneural motor fascicles clumped superficially in the most volar-radial nerve quadrant or, more probably, to the fact that the recurrent thenar branch may exit the carpal tunnel through a separate ligamentous tunnel within the transverse carpal ligament where it may be preferentially or selectively compressed. PMID- 10454286 TI - Inhibition of the human primary motor area by painful heat stimulation of the skin. AB - OBJECTIVE: To prove whether painful cutaneous stimuli can affect specifically the motor cortex excitability. METHODS: The electromyographic (EMG) responses, recorded from the first dorsal interosseous muscle after either transcranial magnetic or electric anodal stimulation of the primary motor (MI) cortex, was conditioned by both painful and non-painful CO2 laser stimuli delivered on the hand skin. RESULTS: Painful CO2 laser stimuli reduced the amplitude of the EMG responses evoked by the transcranial magnetic stimulation of both the contralateral and ipsilateral MI areas. This inhibitory effect followed the arrival of the nociceptive inputs to cerebral cortex. Instead, the EMG response amplitude was not significantly modified either when it was evoked by the motor cortex anodal stimulation or when non-painful CO2 laser pulses were used as conditioning stimuli. CONCLUSIONS: Since the magnetic stimulation leads to transynaptic activation of pyramidal neurons, while the anodal stimulation activates directly cortico-spinal axons, the differential effect of the noxious stimuli on the EMG responses evoked by the two motor cortex stimulation techniques suggests that the observed inhibitory effect has a cortical origin. The bilateral cortical representation of pain explains why the painful CO2 laser stimuli showed a conditioning effect on MI area of both hemispheres. Non-painful CO2 laser pulses did not produce any effect, thus suggesting that the reduction of the MI excitability was specifically due to the activation of nociceptive afferents. PMID- 10454287 TI - Reproducibility of tendon jerk reflexes during a voluntary contraction. AB - OBJECTIVES: The present study explored whether testing tendon jerks during voluntary contraction of the test muscle would improve reproducibility by effectively 'clamping' the excitability of the motoneurone pool at firing threshold. METHODS: Tendon jerk reflexes of soleus, tibialis anterior and vastus lateralis and the soleus H reflex were recorded in 12 healthy subjects at rest and during voluntary contractions of 10-20% of maximum. Recordings were repeated 8-10 times in 5 subjects, in whom reflex symmetry was also determined. RESULTS: Not all tendon jerk reflexes could be recorded at rest, and the variability of latency and amplitude was high. All reflexes could be recorded in each subject during contractions. The latency of tendon jerk reflexes decreased by approximately 2 ms during contractions, but H-reflex latency decreased by only 0.2-0.3 ms. For the tendon jerks, an asymmetry of >3.0 ms at rest and >2.5 ms contracting would be outside 3 SD of the normal mean. In repeat studies, the coefficient of variation of reflex latency was <4% for the tendon jerk. CONCLUSIONS: A voluntary contraction could potentiate the tendon jerk by a number of mechanisms, but the most important is probably enhancement of the excitability of the motoneurone pool. The present techniques should increase the value of tendon reflex testing when assessing possible peripheral nerve, plexus and root disturbances. PMID- 10454288 TI - Standardisation of anal sphincter EMG: high and low threshold motor units. AB - OBJECTIVE: The anal sphincter muscle has a proportion of low threshold motor units (MUs) that are continuously active and other, recruitable high threshold MUs. In standard EMG recordings, motor unit potentials (MUPs) of the later seem to be of higher amplitudes. A quantitative EMG study was performed to assess possible consequences of sampling MUPs at different levels of sphincter activation. METHODS: Fifteen females without uroneurological disorders were studied. After insertion, standard concentric EMG needle was left in the anal sphincter muscle undisturbed for 1 min; then 30 s of the remaining continuous, and 1 min of voluntarily increased EMG activity were recorded on a DAT recorder. MUPs were collected and analysed by 'Multi-MUP' analysis. MUPs analysed during relaxation constituted the 'low threshold MUP pool'. MUPs sampled on activation were checked for those, already sampled during relaxation, (which were discarded), and the remaining MUPs constituted the 'high threshold MUP pool'. Parameters of both MUP pools were compared. RESULTS: High threshold MUPs were found to be significantly larger than low threshold MUPs. CONCLUSIONS: EMG investigator should be aware of the differences of MUPs sampled at various anal sphincter activity levels. For the technique of 'Multi-MUP' analysis sampling at an activity level which provides 3-5 MUPs per detection site would seem practical, providing a standardised approach suitable for comparing normative data with individual findings from most patients. PMID- 10454289 TI - Nerve conduction studies show no exclusive ulnar or median innervation of the ring finger. AB - In 2047 hands of 1260 patients referred for an analysis of their brachialgia, distal antidromic sensory conduction was investigated in the median and ulnar nerves. No median sensory nerve action potentials (SNAPs) were obtained from the ring finger (D4) in 257 hands, while ulnar D4 SNAPs were lacking in 38 hands. An obvious lesion of the relevant nerve could be demonstrated in each of these cases. In all remaining cases, in which D4 SNAPs were obtained with both median and ulnar stimulation, it could be excluded that they were due to co-stimulation. These findings are not in agreement with the occurrence of mononeural innervation of D4 as a physiological variation, which has been reported in some clinical and experimental studies. Comparison of SNAP parameters in 183 hands with increased median nerve distal latencies showed conduction to be more impaired in the fibers innervating D4 than in those supplying D3. PMID- 10454290 TI - Current understanding on the role of retinal pigment epithelium and its pigmentation. AB - Retinal pigment epithelium (RPE) is a monolayer of cuboidal cells that is strategically placed between the rod and cone photoreceptors and the vascular bed of the choriocapillaris. It has many important functions, such as phagocytic uptake and breakdown of the shedded photoreceptor membranes, uptake, processing, transport and release of vitamin A (retinol), setting up the ion gradients within the interphotoreceptor matrix, building up the blood-retina barrier, and providing all transport from blood to the retina and back. This short review focuses on the role of the pigment granules in RPE. Although the biology of the pigment granules has been neglected in the past, they do seem to be involved in many important functions, such as protection from oxidative stress, detoxification of peroxides, and binding of zinc and drugs, and, therefore, serve as a versatile partner of the RPE cell. Melanin plays a role in the development of the fovea and routing of optic nerves. New findings show that the melanin granules are connected to the lysosomal degradation pathway. Most of these functions are not yet understood. Deficit of melanin pigment is associated with age-related macula degeneration, the leading cause of blindness. PMID- 10454291 TI - Effects of 4-tertiary butylphenol on the tyrosinase activity in human melanocytes. AB - Vitiligo is a common dermatological disorder characterized by the development of complete pigment loss from focal lesions that tends to increase in size over time. The etiology of vitiligo, resulting in the disappearance of functional melanocytes from involved skin, is not clearly understood. As a consequence, no satisfactory therapy has been developed. A subtype of vitiligo, termed 'occupational' or 'contact' vitiligo, is increased in individuals who are exposed to materials containing phenolic derivatives, such as 4-tertiary butylphenol (4 TBP). Phenolic derivatives are structurally similar to tyrosine, the initial substrate of tyrosinase in the biochemical synthesis of melanin. Therefore, it has been proposed that phenolic derivatives compete with tyrosine for hydroxylation by tyrosinase and interfere with the completion of melanin synthesis and/or generate cytotoxic intermediates. Our results demonstrated that 4-TBP competitively inhibited both tyrosine hydroxylase and dihydroxyphenylalanine (DOPA) oxidase activities of tyrosinase, i.e., the first two catalytic steps in the biochemical conversion of tyrosine to melanin in cultured human melanocytes. This inhibition occurred at concentrations that did not influence the viability of melanocytes. The tyrosinase activity inhibited by 4-TBP was recovered after removing the treatment. 4-TBP did not affect the function of other enzymes, such as succinate-tetrazolium reductase, acid phosphatase and sulfatase. Since depigmentation occurred without a cytotoxic response after exposure of melanocytes to low concentration of 4-TBP, it is unclear whether the interaction between 4-TBP and tyrosinase leads to the destruction of the melanocytes in 'contact/occupational' vitiligo. PMID- 10454292 TI - Cyclic stretch up-regulates proliferation and heat shock protein 90 expression in human melanocytes. AB - Human skin is repeatedly exposed to mechanical stretching in vivo, but in an ordinary culture of skin cells this prominent feature has been neglected. In order to study whether mechanical stretching plays a role for human melanocytes, we have established a culture technique to mimic this physical stretching: primary cultures of human melanocytes were plated on silicon supports, which undergo a stretching of about 10% of the initial length. After application of repeated stretching and relaxation for 4 days, cell count was significantly (about 40%) enhanced. In addition, we found approximately 2-fold increase in heat shock protein (HSP) 90, both at the protein and mRNA level. HSP 90 is known to bind to Raf-1 and, therefore, may contribute to the Raf-1-MEK (mitogen-activated protein-kinase kinase)-MAPK (mitogen-activated protein-kinase) signaling pathway. Disruption of the Raf-1-HSP 90 multimolecular complex by geldanamycin lead to a considerable decrease in melanocyte cell count. However, geldanamycin did not reverse the stretch-induced growth stimulation. Therefore, the stretch-mediated up-regulation of HSP 90 expression in melanocytes appears to be independent of stretch-mediated growth stimulation. These findings have strong implications for the in vitro cultivation of melanocytes for transplantation purposes. PMID- 10454293 TI - Oculocutaneous albinism in the i6 mutant of the medaka fish is associated with a deletion in the tyrosinase gene. AB - Three mutant alleles (i1, i4, and i5) of the tyrosinase gene in the i locus of the medaka fish Oryzias latipes have hitherto been described, all being associated with transposable element insertion. We have recently identified another allele causing a complete albino phenotype in homozygous carriers and named it i6. Sequence comparison between the tyrosinase gene for the i6 allele (Tyr-i6) and the wild-type gene previously obtained (Tyr-i+) revealed three deletions of 8, 44, and 245 bp. The first two deletions reside in an intron and are differences in the number of tandem tetranucleotide repeats that are polymorphic even among wild-type genes, and, thus, not likely to be responsible for the i6 albino phenotype. The largest deletion spans over the last 180 bp of the second intron and the first 65 bp of the third exon. Because of this deletion, the Tyr-i6 gene lacks the branch point sequence and the acceptor site for the second intron, both being considered to be necessary for normal RNA splicing. Therefore, the 245-bp deletion is likely to be responsible for the albino phenotype. With a mutant gene of this type, unlike ones bearing transposable element insertions, the possibility of reversion mutations to the wild-type would be negligible. Therefore, fish having the i6/i6 genotype should serve as superior recipients for the tyrosinase gene in rescue experiments. PMID- 10454294 TI - Chemical analysis of melanin pigments in feather germs of Japanese quail Bh (black at hatch) mutants. AB - Bh (black at hatch) is a mutation of Japanese quails which causes darkening or lightening of the plumage in heterozygotes or homozygotes, respectively. We chemically analyzed melanin pigments in feather germs of Bh mutant embryos and in feathers of adult animals. Dark brown dorsal feathers of wild-type adult animals had white barrings, but heterozygous ones lacked clear barrings. The feathers of wild-type and heterozygote animals contained both eumelanins and pheomelanins, the latter being more pheomelanic. On the dorsal skin of 10-day old wild-type embryos, longitudinal stripes from black and yellow rows of feather germs developed; two or three longitudinal rows of black feather germs and then two or three rows of yellow feather germs next to the short central feather germs. Heterozygous embryos appeared black in plumage pigmentation, due to the presence of 'gray' feather germs in rows of dorsal feather germs that corresponded to yellow rows in wild-type embryos. Homozygous dorsal feather germs did not develop the black and yellow longitudinal stripes, but were brown. Chemical analysis showed that embryos of each genotype contained both eumelanins and pheomelanins in the feather germs; however, the eumelanin content in homozygous feather germs was very low. These results suggest that the Bh mutation causes pheomelanic changes in feathers of quails. PMID- 10454295 TI - Pheomelanin as a binding site for drugs and chemicals. AB - Certain drugs and chemicals, such as chloroquine, chlorpromazine, and 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP), are bound to melanin and retained in pigment cells for long periods. This specific retention in pigmented tissues can cause adverse effects in the skin, eye, inner ear, and pigmented nerve cells of the substantia nigra of the brain. To date, all studies have been focused on eu- and neuromelanin. In the present study, we show that chloroquine, chlorpromazine, chlomipramine, paraquat, acridine orange, and nickel, which are bound to eumelanin, also bind to synthetic pheomelanin, but the binding to pheomelanin is lower. The binding varied with the cysteine content and pH, and the results indicate that the binding is complex and includes ionic interactions. In addition, we have shown that these substances also bind to synthetic thiourea containing melanin, but to quite a low extent. We also present a microautoradiographic study on the binding of 14C-chloroquine to natural pheomelanin in vivo in yellow mice C57BL (Ay/a). Black (C57/BL) and albino (NMRI) mice were used as controls. The autoradiography demonstrated a pronounced uptake of chloroquine in the hair follicles and the dermal melanocytes in the ear of yellow mice, which was comparable to the corresponding accumulation of label in black mice. In the albino mouse, the uptake was lower and more homogeneously distributed in the skin. These results suggest that the toxicological risks of melanin-related adverse effects are applicable to persons with a high content of pheomelanin in the skin and hair. PMID- 10454296 TI - Ommochrome deficiency in an albino strain of a terrestrial isopod, Armadillidium vulgare. AB - In order to clarify the cause of ommochrome deficiency in an albino strain of the terrestrial isopod, Armadillidium vulgare, levels of xanthommatin, 3 hydroxykynurenine, 3-hydroxyanthranilic acid and tryptophan in whole body extracts of the albino and the wild type individuals were determined together with enzyme activities of kynurenine-3-hydroxylase, kynureninase and tryptophan 2,3-dioxygenase. Xanthommatin could not be detected in the albinos. The levels of 3-hydroxykynurenine and 3-hydroxyanthranilic acid were determined by high performance liquid chromatography (HPLC) with electrochemical detection and were markedly low in the albinos compared with the wild type individuals. In contrast to those, the tryptophan levels determined by HPLC with fluorescence detection did not differ significantly between the two phenotypes. In the albino A. vulgare, kynurenine-3-hydroxylase activity was lower and kynureninase activity was higher than in the wild type, although the differences were not statistically significant. Tryptophan-2,3-dioxygenase activity in the albinos was less than 10% that in the wild type. Thus, ommochrome deficiency in the albino A. vulgare is considered to be caused by the extremely low activity of tryptophan-2,3 dioxygenase. PMID- 10454297 TI - Exaggerated heart rate oscillations during two meditation techniques. AB - We report extremely prominent heart rate oscillations associated with slow breathing during specific traditional forms of Chinese Chi and Kundalini Yoga meditation techniques in healthy young adults. We applied both spectral analysis and a novel analytic technique based on the Hilbert transform to quantify these heart rate dynamics. The amplitude of these oscillations during meditation was significantly greater than in the pre-meditation control state and also in three non-meditation control groups: i) elite athletes during sleep, ii) healthy young adults during metronomic breathing, and iii) healthy young adults during spontaneous nocturnal breathing. This finding, along with the marked variability of the beat-to-beat heart rate dynamics during such profound meditative states, challenges the notion of meditation as only an autonomically quiescent state. PMID- 10454298 TI - Spatial features in body surface potential maps of patients with ventricular tachyarrhythmias with or without coronary artery disease. AB - Body surface potential maps (BSPM) from patients with coronary artery disease or no structural heart disease were analyzed with respect to their spatial features and QT/QTc dispersion in order to determine whether BSPM allows identification of patients with ventricular fibrillation. QRST integral maps and QT/QTc dispersion were acquired from simultaneous recordings of 62 ECG leads during sinus rhythm in patients with idiopathic ventricular fibrillation (n=13), ventricular fibrillation and coronary artery disease (n=22), coronary artery disease without ventricular fibrillation (n=21) and healthy controls (n=18). The Karhunen-Loeve transformation was applied to reduce the dimensionality of the data matrix of the QRST map to eight coefficients. Linear discriminant analysis allowed discrimination between idiopathic ventricular fibrillation patients and controls with high sensitivity (85%) and specificity (89%). However, discrimination between coronary artery disease patients with or without ventricular fibrillation was poor (68% and 67%, respectively). QTc dispersion calculated from BSPM was longer in idiopathic ventricular fibrillation patients than in controls (99+/-30 ms vs 70+/-14 ms, P=0.009) in contrast to QTc dispersion taken from 12-lead ECG (53+/-21 ms vs. 47+/-12 ms, P=n.s.). No significant difference was noted for coronary artery disease patients with or without ventricular fibrillation. In conclusion, repolarization disturbances detected by BSPM allow identification of ventricular fibrillation patients without structural heart disease. However, our results do not suggest a major impact of QT/QTc dispersion or QRST integral mapping for identification of ventricular fibrillation patients with coronary artery disease. PMID- 10454299 TI - D allele of the angiotensin-converting enzyme gene is a risk factor for secondary cardiac events after myocardial infarction. AB - We retrospectively examined the relationship between the genotype of the angiotensin-converting enzyme (ACE) gene or the 5,10-methylenetetrahydrofolate reductase (MTHFR) gene, and the secondary cardiac events after myocardial infarction. The study population consisted of 176 patients (ACE genotype: deletion homozygote (DD)=20, insertion/deletion heterozygote (ID)=91, insertion homozygote (II)=65; MTHFR genotype: valine homozygote (VV)=37, valine/alanine heterozygote (VA)=71, alanine homozygote (AA)=68) with acute or recent myocardial infarction at the start of the follow-up. We defined the occurrence of cardiac death, recurrent myocardial infarction, or admission due to unstable angina as the endpoint. Cardiac events related coronary intervention were excluded from the endpoints. During the follow-up (1903+/-1414 days), four patients had cardiac death, 12 patients had recurrent myocardial infarction and 13 patients had admission due to unstable angina. A Cox analysis revealed that the endpoints were significantly associated with diabetes mellitus (RR=4.423), total cholesterol level (RR=1.025) and the genotype of the ACE gene (RR=4.490). The ID or DD genotype of the ACE gene was associated with higher occurrence of the endopoints. The MTFHR gene was not associated with the endopoint. The present results suggest that the presence of the deletion allele of the ACE gene may be a risk factor for secondary cardiac events after myocardial infarction. PMID- 10454300 TI - Changes in renal function with percutaneous transluminal coronary angioplasty. AB - There is concern about adverse effects on renal function in patients with prolonged cardiac intervention procedures, when contrast media is used. To investigate this further we studied changes in renal function in 104 patients (79 male, 25 female; mean age 59.2, SD 9.8) undergoing routine elective percutaneous transluminal coronary angioplasty (PTCA), where 28 (27%) patients had concomitant stent implantation. There was associated diabetes in 15 patients (14%) and previous hypertension in 44 (blood pressure >160/90 mmHg, 44%). None of the patients were known to have congestive heart failure at the time of procedure or chronic renal failure (defined as serum creatinine >200 pmol/l). There was no significant change in mean serum urea pre- and post-PTCA (mean change -0.04 mmol/l, paired t-test P=0.90). However, there was a small rise in serum creatinine pre- and post-PTCA of borderline significance (mean change +5.8 micromol/l, P=0.051). Of the whole cohort, 65 patients (63%) had a rise in mean serum creatinine, whilst 45 (43%) showed a rise in serum urea levels. This deterioration in renal function was related to a difference in the procedure duration, but there were no statistically significant differences in mean age or volume of contrast media (Iopamide 340) between patients with or without deterioration in renal function. Patients with a rise in serum creatinine had lower baseline (pre-PTCA) serum urea and serum creatinine levels. In patients undergoing stent implantation, there was a higher quantity of contrast media, screening time and procedure duration. There were no significant differences in age, pre-PTCA serum urea and creatinine levels, and mean change in serum urea or creatinine levels in patients with and without stent usage. Whilst severe renal dysfunction following PTCA is uncommon, we suggest that some caution is necessary during PTCA or other cardiac interventions where more complex or prolonged procedures necessitating large volumes of contrast media use. PMID- 10454301 TI - Persistent dissection of carotid artery in patients operated on for type A acute aortic dissection--carotid ultrasound follow-up. AB - Over a period of 5 years, 124 patients were operated on at the National Institute of Cardiology in Warsaw for acute aortic dissection, 27 of whom died. The 97 patients discharged from the hospital were included in the present analysis. The age of the patients ranged from 25 to 73 years with a mean of 50+/-10 years. Ultrasound examination of carotid arteries was performed with the patient lying on his back using a Toshiba 340A color Doppler system with a linear array probe of 7.5 MHz. Of the 97 patients examined, 15 (15%) had a dissection of at least one of the common carotid arteries (CCA). Two had Marfan syndrome. In 11 patients the dissection involved the right common carotid artery and in four it involved both the left and right common carotid arteries. The flow in the true lumen of CCA and ICA was preserved in all patients and the degree of narrowing ranged from 30 to -70%. Only one of the 15 patients with CCA dissection had an ipsilateral neurological deficit which was already present before the aortic aneurysm operation. Ultrasound follow-up was performed in all patients with the CCA dissection found on first examination. The mean duration of follow-up was 21 months. In 14 patients the degree and extent of the dissection as well as the narrowing of the true lumen was comparable, and in one patient the false channel closed spontaneously. During follow-up there were no new major neurological events despite the persistence of the CCA dissection with different degrees of narrowing of the true lumen. Doppler ultrasound examination of the carotid arteries can supply additional information about the extent of the dissection, and help to assess the flow in the persisting 'double channel' common carotid artery during the follow-up of patients. PMID- 10454302 TI - Regression of ventricular repolarisation inhomogeneity after aortic bileaflet valve replacement in patients with aortic stenosis. AB - BACKGROUND: Aortic valve replacement relieves mechanical outflow obstruction in patients with aortic stenosis. However, there is limited information on whether aortic valve replacement can provide regression of ventricular repolarisation inhomogeneity. OBJECTIVES: To determine whether aortic valve replacement can provide regression of ventricular repolarisation inhomogeneity in patients with aortic stenosis after bileaflet aortic valve replacement. METHODS: We studied the changes of electrocardiographic QT or QTc intervals and QT or QTc dispersions of 71 patients with severe aortic stenosis and angiographically insignificant coronary lesions (<50% in diameter) before and after valve replacement (6+/-3 days after operation). Seventy-one healthy control subjects, matched for age and sex, served as control subjects. Twelve-lead electrocardiograms and echocardiographic examinations were measured before and after surgery. The QT interval was corrected for heart rate using the standard Bazett formula. QT dispersion was defined as the difference between maximal and minimal QT interval measurements occurring among any of the 12 leads on a standard electrocardiogram. QTc dispersion was calculated in a manner similar to QT dispersion. No subject had fewer than nine measurable leads. RESULTS: Left ventricular systolic blood pressure, pressure gradient across aortic valve, left ventricular mass index, and systolic wall stress were significantly reduced after valve replacement compared with before valve replacement. The QT interval significantly decreased from 425+/ 38 ms to 398+/-32 ms after replacement (P<0.0001). The QTc dispersion significantly decreased from 62+/-25 ms to 32+/-13 ms after replacement (P<0.0001). The value of QT or QTc dispersion after replacement was similar to that in controls. Univariate analysis revealed that QTc dispersion was significantly only correlated with left ventricular mass index (r=0.236, P=0.05). Multivariate analysis revealed that the best predictor of QTc dispersion was sex and left ventricular mass index (P=0.008 and 0.005, respectively). CONCLUSIONS: Our study demonstrated a favorable consequence of aortic valve replacement distinct from hemodynamic improvement. Patients with aortic stenosis before valve replacement have abnormal prolonged QT or QTc intervals and increased QT or QTc dispersions. After successful valve replacement left ventricular mass index regressed and QT or QTc intervals and QT or QTc dispersions were normalized. These findings warrant further investigation in a large trial and long-term follow-up for clinical implications. PMID- 10454303 TI - Is there any effect of Ramadan fasting on acute coronary heart disease events? AB - OBJECTIVE: We investigated whether Ramadan fasting, which concerns billions of Muslims in the world, has a negative effect on coronary heart disease patients. METHODS: Patients who were hospitalized at Emergency Center of Ankara Numune Hospital were evaluated retrospectively between the years 1991 and 1997. Patients with acute coronary heart disease events, who were hospitalized and those who died of this disease within the period of before, during, and after Ramadan were evaluated and the ratio of these cases in the total number of patients was determined along with mortality rates. RESULTS: In all the years of the study the number of cases with acute coronary heart disease events was significantly lower in Ramadan than before or after Ramadan (P=0.03). But, the ratio of this population to all patients was not statistically significant between the periods (P>0.05). In conclusion, we speculate that Ramadan fasting does not increase acute coronary heart disease events. We believe that further prospective studies should provide an opportunity to examine the relation of fasting to coronary events. PMID- 10454304 TI - Mental stress test is an effective inducer of vasospastic angina pectoris: comparison with cold pressor, hyperventilation and master two-step exercise test. AB - BACKGROUND: Cold pressor, hyperventilation and exercise stress tests were usually used for inducing an angina attack in patients with vasospastic angina pectoris. We induced vasospastic angina attack using the mental calculation stress test, and compared the results with those using other stress tests. SUBJECTS AND METHODS: Subjects were 29 patients with vasospastic angina pectoris. Their ages were 60.8+/-8.4 years. Coronary vasospasm was induced by an acetylcholine infusion test during coronary angiography. The mental stress test was performed as follows; after memorizing six digits numbers, they repeated these numbers in reverse for 5 min, and performed serial subtraction of 17 from 1000 for 5 min. Blood pressure, heart rate and ECG were recorded every 1-5 min during the mental stress test. The serum concentrations of epinephrine and norepinephrine were measured before and during the mental stress test. We compared these results with those obtained using cold pressor, hyperventilation and the Master two-step exercise stress test. RESULTS: (1) Eight of the 29 patients (28%) showed ischemic ST-T change, which was caused by the mental stress test. (2) The increase in norepinephrine was greater in patients with an ST-T change than without an ST-T change (0.11+/-0.06 vs. 0.04+/-0.04 ng/ml, P<0.01). (3) The incidence of the ST-T change caused by the mental stress test (28%) was similar to the cold pressor test (27%) and greater than that caused by the hyperventilation test (13%). The incidence of ST-T change caused by the Master two-step test was 55%. CONCLUSIONS: The mental stress test is an effective inducer of vasospastic angina attack, and attack may be induced by neurohumoral vasoconstrictive reflex and/or increased left ventricular afterload. PMID- 10454305 TI - Heparin potentiates collateral growth but not growth of intramyocardial endarteries in dogs with repeated coronary occlusion. AB - Heparin accelerates coronary collateral development in various animal models of myocardial ischemia. The purpose of this study was to clarify the beneficial effect of heparin on canine collateral development. Seventeen adult mongrel dogs were instrumented for measurements of a subendocardial segment length in the central area perfused by the left circumflex coronary artery, its flow, and left ventricular pressure. A pulsed Doppler flow probe and an externally inflatable pneumatic occluder were placed around the proximal circumflex artery. After the recovery from surgery, 2-min circumflex coronary artery occlusions were repeated eight times at 58-min intervals daily. After excluding seven dogs with well developed preexisting collateral circulation, ten dogs were randomized into two groups with (n = 5) and without (n = 5) heparin treatment. The total occlusion time until adequate collateral development (an index of collateral growth) was 164+/-34 (SD) min in dogs with heparin treatment, being significantly less than 289+/-49 min in dogs without heparin (p<0.01). In contrast, the extent of the reduction in resting blood flow of the repeatedly occluded circumflex artery (an index of neovascularization toward the ischemic area) was comparable in dogs with and without heparin (15.4+/-12.4% vs. 21.1+/-13.6%, p=NS). Heparin promotes nonsprouting angiogenesis (arteriogenesis) of preformed collateral vessels but not neovascularization toward the ischemic area in dogs with brief repetitive coronary occlusions. PMID- 10454306 TI - Intraventricular conduction delay: a prognostic marker in chronic heart failure. AB - Chronic heart failure (CHF) is associated with high mortality, and there are several established clinical and laboratory parameters that predict mortality in CHF. The purpose of this study was (a) to identify the best ECG parameter that predicts mortality, (b) to evaluate the prognostic marker of ECG against well established indicators of prognosis. Relevant data from 241 CHF patients were analysed retrospectively. Cardiopulmonary exercise testing and radionuclide ventriculogram were also performed where possible. The mean follow-up period was 31 months. On univariate analysis by the Cox proportional Hazard method, intraventricular conduction delay (IVCD) [P<0.0001, hazard ratio 1.017 (1.011 1.024)] and QTc [P<0.0001, hazard ratio 1.012 (1.006-1.017)] were identified as predictors of mortality. On bivariate analysis, IVCD and MVO2 were better predictors when combined together. A model based on multivariate analysis showed that IVCD, MVO2 and left ventricular ejection fraction (LVEF) were the best predictors of mortality. The addition of plasma sodium, age and NYHA class had no added benefit on the predictive power of the model. Further analysis of IVCD and QTc showed that, for different cut-off values, IVCD is better than QTc, and that there is a graded increase in mortality with increasing value of IVCD. We have found that IVCD is an important ECG predictor of prognosis in patients with CHF. PMID- 10454307 TI - The clinical impact of dynamic intraventricular obstruction during dobutamine stress echocardiography. AB - We selected 73 consecutive patients without myocardial-infarction, hypertrophic cardiomyopathy or hypertension complaining of effort chest discomfort/dyspnoea, and/or reporting exercise ischaemic ECG changes, and submitted them to simultaneous dobutamine stress echocardiography (DSE) and 99mTc tetrofosmin SPECT (T SPECT) and to coronary angiography to evaluate the clinical impact of intraventricular obstruction (IVO) during dobutamine infusion. Sixteen patients (22%, 7 males, mean age+/-SD 63+/-8 years, group 1) developed IVO (mean CW Doppler velocity+/-SD: 3.8+/-1.0 m/s) and 57 (41 males, mean age+/-SD 63+/-10 years, group 2) did not. The two groups had similar incidence of angina and ischaemic ECG changes at exercise tolerance test. DSE did not demonstrate wall motion abnormalities in any group 1 patient while T SPECT showed a perfusion defect in the only one with coronary artery disease (CAD). DSE reproduced symptoms in a higher percentage of patients with than without IVO, while there was no statistical difference in the reproduction of ischaemic ECG changes, despite CAD prevalence was much lower in group 1. Group 1 patients remained asymptomatic on beta-blockers at 12-month follow-up. Dobutamine-induced IVO, by reproducing symptoms, suggests that IVO plays a role in the clinical setting in patients without CAD complaining of unexplained reduced effort tolerance who should undergo DSE. PMID- 10454308 TI - Historical note: the first control study on treatment of angina pectoris--Inga Lindgren in 1950. PMID- 10454309 TI - Historical note: Inga Lindgren's paper in 1947 on the effect of cutaneous precordial anaesthesia on the electrocardiogram in patients with angina pectoris and coronary occlusion. PMID- 10454310 TI - Intrathoracic gossypiboma: magnetic resonance features. AB - Gossypiboma, a term used to describe a mass within the body composed of a cotton matrix which usually refers to a retained surgical sponge, is a rare complication of cardiac surgery. We report one case revealed by the recurrence of the anginal symptoms after successful myocardial revascularisation surgery. The preoperative diagnosis was suspected by a combination of imaging techniques and subsequently confirmed by surgical removal. We review the literature to summarise the diagnostic and therapeutic features of gossypibomas. PMID- 10454311 TI - Aortico-right ventricular tunnel. AB - A 2-year-old child with aortico-right ventricular tunnel is reported for its rarity. The right coronary artery originated from the distal end of the tunnel. The frequent occurrence of coronary artery origin abnormality with this anomaly is highlighted. PMID- 10454312 TI - Atrial standstill in a case of Kugelberg-Welander syndrome with cardiac involvement: an electrophysiologic study. AB - A patient with Kugelberg-Welander syndrome associated with junctional rhythm and restrictive cardiomyopathy is presented. An electrophysiologic study was performed. Persistent atrial standstill was demonstrated by detailed right atrium and coronary sinus mapping and failure of capture of both right atrium and coronary sinus pacing. A prolonged junctional recovery time, normal HV interval and normal pacing threshold of the right ventricle were also noted. The patient was successfully treated with cardiac pacing and diuretics. PMID- 10454313 TI - Acute myocardial infarction in a young patient during an exacerbation of ulcerative colitis. PMID- 10454314 TI - Buccal fat pad pedicle flap for midface augmentation. AB - Midface aging is characterized by soft-tissue ptosis with loss of cheek projection. Subperiosteal midface lifts may reposition the soft-tissue mounds and improve the tear trough, but may not fill the lateral cheeks in patients with significant jowls or poor bony support. Correction with alloplastic implants is helpful, but may not be accepted by many patients. During subperiosteal midface lifts, the author often excises Bichat's fat pad to decrease the jowl and to diminish face fullness. He has modified this approach and used a vascularized Bichat's fat flap to aid lateral cheek projection while still improving lower face fullness and the jowl. For the last 4 years, close to 150 patients undergoing subperiosteal midface lifts have had vascularized Bichat's fat pad flaps. The jowls were marked preoperatively. All patients had complete cheek undermining either through a buccal sulcus incision or through a crow's-foot incision, or through a muscle-sparing limited lower blepharoplasty incision. Bichat's fat pad is identified in its pocket medial to the masseter tendon. Mobilization of Bichat's fat pad is done by blunt dissection, preserving its thin fascial envelope. The "hernial saclike" pocket, excluding Stensen's duct and the buccal branches of the facial nerve, is identified and protected. Suspension is accomplished by fixation with 3-0 polydioxanone sutures either to the temporalis fascia (via the temporal incisions), to the arcus marginalis, or to the suborbicularis oculi fat pad. Fixation technique is dependent on where the fat pad is needed and the surgeon's preference. Fat pad repositioning is accomplished with a minor learning curve. The most common problems are tearing of the fat pad during fixation and temporary numbness of the long buccal nerve. Attention to leaving the capsule intact and gentle handling is essential to fixation. Nevertheless, in some patients with poor-quality fat pads, fixation is extremely difficult. Four-year results have been excellent. Further studies with magnetic resonance imaging of postoperative patients are necessary to assess longevity. Bichat's fat pad provides autologous vascularized tissue for midface fill. Placement may be lateral for cheek augmentation or medial for deep nasolabial folds. Jowl improvement also occurs with the removal of Bichat's fat pad from its pocket. PMID- 10454315 TI - The "parasite" TRAM flap for autogenous tissue breast reconstruction in patients with vertical midabdominal scars. AB - Abdominal scars play an important role in risk factors in transverse rectus abdominis musculocutaneous (TRAM) flap breast reconstruction. In particular, vertical midline scars are a difficult problem to solve. Traditional techniques include the use of a single hemiflap (which may be insufficient to achieve an adequate volume), the transfer of a double-pedicle flap (which causes major trauma to the abdominal wall), or more complicated procedures such as two free hemiflaps. Since 1991 the authors have used an innovative technique to improve vascularity in the contralateral side of a standard unipedicled TRAM flap. They call this flap the recharged TRAM flap. By means of their technique, the retrograde flow coming from the deep inferior epigastric vessels raised in continuity with a superiorly pedicled flap is used to "recharge" the contralateral rectus muscle harvested as a free flap. On the basis of hemodynamic studies, this procedure was carried out in patients with vertical midabdominal scars. This flap was named the parasite flap because the free unit survives on the vascular source of the pedicled unit-the superior epigastric artery supplying both flaps in a retrograde fashion. Sixteen patients with vertical midabdominal scars underwent this procedure. Total flap survival was observed in 15 patients. One patient developed a partial flap necrosis and 1 patient developed abdominal bulging on the pedicled side. According to the surgeons' evaluation, aesthetic outcome was considered to be good to excellent in all patients. PMID- 10454317 TI - Breast reconstruction following excision of phylloides tumor. AB - There are few papers published on breast reconstruction after excision of phylloides tumor. Six patients who had reconstruction of the breast following excision of phylloides tumor are described. All underwent wide excision or subcutaneous mastectomy followed by immediate or delayed reconstruction with implants or autologous tissue. The mean follow-up was 5 years (range, 2.5-7 years). One patient died of metastases; the others survived without evidence of recurrence. The etiology, incidence, diagnosis, and treatment of these tumors are discussed. The aesthetic results in these patients is also described. PMID- 10454316 TI - Dilute adrenaline infiltration and reduced blood loss in reduction mammaplasty. AB - The aim of this review was to assess the use of dilute adrenaline infiltration in reduction mammaplasty and to determine whether it had any associated complications. The closed technique for adrenaline infiltration was used with no reported infection. One hundred breast reductions in 50 patients were compared by dividing them into two groups of 25 patients each. Group A had 1:500,000 adrenaline in normal saline infiltration; group B did not. Both groups were matched equally for age and general health. Results showed that blood loss was less for group A when measured by the fall in postoperative hemoglobin (2.5 g per deciliter vs. 3.5 g per deciliter). This was statistically significant (p < 0.05). There was no significant difference in postoperative drainage (group A, 158 ml; group B, 182 ml). Group A required fewer blood transfusions (two vs. eight), without the risk of increased complications. Blood transfusions were given in the earlier part of the study, but currently are rarely needed. Adrenaline infiltration at this dilution is virtually free from any side effects. It decreases intraoperative blood loss and facilitates the operation without the need for blood transfusion. PMID- 10454318 TI - Deepithelialization prior to onlay grafting using the UltraPulse carbon dioxide laser. AB - When used in a defoeused mode, the UltraPulse carbon dioxide laser causes skin vaporization at a subepidermal level with minimal surrounding thermal destruction, allowing early reepithelialization from retained skin appendages. This attribute is of paramount importance if only skin resurfacing is desired. However, because the effect essentially is total removal of the epidermis, such a sequela can also be used advantageously to prepare a vascularized dermal bed prior to onlay of composite grafts or secondary overgrafting. This skin deepithelialization can be achieved in a bloodless field, permitting constant visualization of the depth of penetration, and an integrated computerized pattern generator scanner allows rapid, precise, and consistent desurfacing to the desired level. This preliminary investigation demonstrates that successful overgrafting is possible on a viable dermal plane exposed by the UltraPulse carbon dioxide laser, as another example of its role in skin deepithelialization. PMID- 10454319 TI - Velopharyngeal anthropometric analysis with MRI in normal subjects. AB - Magnetic resonance imaging (MRI) is a new method of evaluating velopharyngeal dysfunction. The anatomy of the palate and velopharyngeal closure can be visualized with MRI. Also, fast scanning with turboflash sequences allows surgeons to evaluate the speech mechanism during phonation. The dimensions of the velopharynx are another important factor in velopharyngeal closure, together with the functional performance of the soft palate during phonation. During this study, velopharyngeal anthropometric analyses were performed in cerebral MR images of 136 male and 141 female normal speakers who underwent cerebral MRI for reasons other than speech problems. Age-dependent changes in velopharyngeal dimensions were evaluated. Although the growth of the soft palate was continuous throughout life, hard palate growth ceased by age 15. By using anthropometric data available from MR images taken on normal subjects for other reasons, a large database of age-, gender-, and race-specific data on palatal and pharyngeal morphology was acquired. Comparative data of noncleft subjects in multiple age groups were obtained with MRI. PMID- 10454320 TI - Splitting advancement genioplasty: a new genioplasty technique. AB - A new genioplasty technique has been described and performed on 16 patients since 1995. The technique has been developed to avoid some undesired results of the current osseous genioplasty techniques and to achieve a more natural appearance in advancement genioplasty. According to the authors' technique, a rectangular part of the outer table of the mentum is split away from the mandible, and is advanced and fixated to the mandible. This technique can be used for advancement cases but not for reduction genioplasty. This technique was performed on 16 patients with only minor complications, including one case of wound dehiscence, one hematoma, and one case of osteomyelitis, which was managed with systemic antibiotic therapy. Aesthetic results were found to be satisfactory according to an evaluation by the authors. When the results were evaluated using pre- and postoperative photos, lip position and projection of the mentum were found to be natural in shape appearance. During the late postoperative period, the new bone formation between the advanced segment and the mandible was demonstrated radiographically. Advantages of the technique include having more contact surfaces for bony healing, a natural position of the lower lip, more natural projection of the mentum, tridimensional movement of the mentum, and improvement in the soft tissue of the neck. The disadvantages of the technique are the potential risk of infection due to dead space from the advancement, manipulation problems during surgery, and possible mental nerve injury. Splitting advancement genioplasty was found to be a useful technique for advancement genioplasty. Splitting advancement genioplasty is a more physiological osteotomy technique than most of osseous genioplasty techniques. PMID- 10454321 TI - "Outbreak" of hand injuries during Hajj festivities in Saudi Arabia. AB - Pediatric hand surgeons in Europe and North America are aware of the yearly "outbreak" of pediatric hand injuries on Halloween from carving pumpkins. This study reports another yearly "outbreak" of hand injuries--in Saudi Arabia from slaughtering sheep. During the 3 days of festivities following the Hajj ceremony, hundreds of thousands of sheep are slaughtered. Over 4 consecutive years, 298 patients attended the emergency room of Riyadh Central Hospital with injuries related to the slaughtering of animals sustained during the 3 days of festivity. Almost three quarters of patients (73%) were injured on the first day of festivity. The majority (92%) were nonprofessionals. Children accounted for 6.7% of patients. In adults, the female-to-male ratio was 1:7. The most common mechanism of injury was a knife cut (80.9%). Almost three quarters of injuries (73.5%) affected the hands, with more involvement of the left than the right hand. Only 7.7% of patients with hand injuries were admitted to the hospital for treatment. It was concluded that implementing safety measures while slaughtering, and educating the general population would be important in the prevention of these hand injuries in Saudi Arabia. PMID- 10454322 TI - Frozen section analysis in the management of skin cancers. AB - Frozen section analysis is used routinely to ensure complete removal of basal cell and squamous cell carcinomas of the skin, but the current emphasis on controlling costs raises the question as to which lesions should be evaluated with frozen section histology. A retrospective study of the clinical records and pathological reports of 51 patients selected randomly from a total of 225 patients was undertaken to determine the overall impact of frozen section analysis on the surgical management of skin cancers at this institution. The results indicate that frozen section analysis was performed on 76% of the 277 lesions evaluated, and that the results of frozen section examination were 91.1% accurate in detecting the presence or absence of tumor involvement at the surgical margins. The data also showed that the surgeons were able to estimate the margin of the skin tumor clinically and remove it entirely during the first excision 91.1% of the time. The lesions that had not been removed completely with the initial excision were those located on the periorbital region, forehead, and cheeks; were recurrent lesions; or were lesions that required more involved reconstruction than primary closure. This study shows that frozen section analysis is a valuable tool in selected situations, but routine use is not indicated for the majority of basal and squamous cell carcinomas of the skin. PMID- 10454323 TI - Introduction of cremaster muscle chamber technique for long-term intravital microscopy. AB - This study evaluated the microcirculatory hemodynamics of a new chamber implantation technique. The cremaster muscle island flap was employed. Seventeen male Sprague-Dawley rats were studied in two groups. In the control group, the standard cremaster muscle preparation with no chamber (N = 8) was used. After flap isolation, the muscle was preserved in the medial border of the hind limb and removed for observation after 24 hours. For the chamber group, the chamber was implanted after muscle isolation, and measurements were made 30 minutes postoperatively and at 24, 48, and 72 hours. The variables measured were microvessel diameter, red blood cell velocity, number of perfused capillaries, and the number of rolling, sticking, and transmigrating leukocytes in the postcapillary venules. The chamber group had a significantly greater number of perfused capillaries at 24 hours compared with controls (p < 0.05). The other variables did not differ significantly between groups at 24 hours. We can conclude that this cremaster muscle chamber model for chronic in vivo studies proved to be equal to the classic cremaster muscle preparation for chronic microcirculatory measurements for at least 24 hours. PMID- 10454324 TI - Patterns of flap loss related to arterial and venous insufficiency in the rat pedicled TRAM flap. AB - Vascular supply to the contralateral portion of the conventional transverse rectus abdominis musculocutaneous (TRAM) flap (zone IV) may become compromised, resulting in partial flap loss and requiring segmental excision. The etiology of this necrosis is not clear. This study determines skin necrosis patterns on a superiorly pedicled caudal TRAM flap during conditions of venous and arterial insufficiency, and determines whether cutaneous venous outflow can sustain a flap with venous insufficiency. Twenty-eight adult male Sprague-Dawley rats underwent superior pedicled TRAM flap elevation, and the zones were marked on the skin paddle. The animals were divided into four groups: control (group A, N = 6), arterial ligation (group B, N = 6), venous ligation (group C, N = 8), and venous ligation with cutaneous venous outflow (group D, N = 8). After 10 days, the skin paddle was photographed and the areas of necrotic skin were measured. Results showed that group B (selective arterial ligation) had 51.7 +/- 2.8% and 40.0 +/- 2.0% skin necrosis in zones I and II respectively. Zone I necrosis was significantly greater in group B compared with the control (p < 0.05). Group C (selective venous ligation) resulted in 73.8 +/- 16.4% and 93.8 +/- 33.4% skin necrosis in zones III and IV respectively. This necrosis was significantly greater compared with the control (p < 0.001). Group D rats' lateral skin necrosis compared significantly less with group C (p < 0.001). These results demonstrate that the patterns of flap necrosis in rat TRAM flaps with poor arterial inflow differ from those with venous stasis. Necrosis of the contralateral portion (zone IV) of human TRAM flaps may be related to problems with venous stasis; thus, a cutaneous venous outflow may prevent this problem. PMID- 10454326 TI - Expression of transforming growth factor beta 1, 2, and 3 proteins in keloids. AB - Keloids represent a pathological response to cutaneous injury, creating disfiguring scars with no known satisfactory treatment. They are characterized by an excessive accumulation of extracellular matrix, especially collagen. Transforming growth factor beta (TGF-beta) has been implicated in the pathogenesis of keloids. The three TGF-beta isoforms identified in mammals (TGF beta1, -beta2, and -beta3), are thought to have different biological activities in wound healing. TGF-beta1 and TGF-beta2 are believed to promote fibrosis and scar formation, whereas TGF-beta3 has been shown to be either scar inducing or reducing, depending on the study. The aim of this study was to characterize expression of TGF-beta isoforms in keloids at the protein level using Western blot analysis. The authors found that TGF-beta1 and -beta2 proteins were at higher levels in keloid fibroblast cultures compared with normal human dermal fibroblast cultures. In contrast, the expression of TGF-beta3 protein was comparable in both the normal (N = 3) and keloid (N = 3) cell lines. These findings, demonstrating increased TGF-beta1 and -beta2 protein expression in keloids relative to normal human dermal fibroblasts further support the roles of TGF-beta1 and -beta2 as fibrosis-inducing cytokines. PMID- 10454325 TI - Effects of VEGF administration following ischemia on survival of the gracilis muscle flap in the rat. AB - The incidence of free flap transplantation failure is only 3% to 5%, yet still occurs in cases in which the flap suffers prolonged ischemia. The purpose of the current study was to determine the effects of vascular endothelial growth factor (VEGF)--a potent angiogenic agent with a suspected role in the protection of endothelium--on flap survival in a model of ischemia-reperfusion injury. The model chosen was the rat gracilis muscle flap. A total of 36 adult male Sprague Dawley rats were divided into three groups (N = 12). One experimental group received VEGF treatment and the other received heparin. A third group was treated with saline and served as the control. The gracilis muscle flap was dissected and isolated based on a vascular pedicle originating at the femoral vessels. Following 3.75 hours of ischemia, induced by clamping the femoral vessels, either VEGF, heparin, or saline was infused directly into the pedicle of the flap via a cannula. The flaps were evaluated both grossly and histologically after 72 hours of reperfusion. Eleven of the 12 flaps from the VEGF group survived, whereas the survival rate was 6 of 12 and 5 of 12 flaps for the heparin- and saline-treated groups respectively. Flap survival was significantly greater in the VEGF-treated group compared with the heparin- and saline-treated groups (p < 0.025, p < 0.01 respectively). Furthermore, there was no significant difference between the heparin and saline groups. These results indicate that VEGF plays a role in reducing the damage that occurs in ischemia-reperfusion injury, and that the use of VEGF holds promise as a potential therapy for increasing flap survival. PMID- 10454327 TI - Effect of TGF-beta2 on proliferative scar fibroblast cell kinetics. AB - Keloids, hypertrophic scars, and burn hypertrophic scars are all forms of proliferative scarring characterized by overabundant matrix formation. Recently these dermal proliferative disorders have been linked clinically to the cytokine transforming growth factor beta (TGF-beta), and in vitro tests have shown it to be responsible for the activation of fibroblasts and their production and deposition of collagen. Using an established in vivo animal model of proliferative scarring, the effects of this cytokine, specifically the isoform TGF-beta2, on these scars were examined. Proliferative scar specimens were implanted into athymic, asplenic nude rats and isolated in sandwich island flaps based on the superficial inferior epigastric pedicle. After establishment of the transferred flap, the scars were injected with varying doses of TGF-beta2 or vehicle for 5 consecutive days and then again on days 10, 15, and 20. The specimens were measured weekly during the period of dosing, and a biopsy was acquired on days 30 and 60. Fibroblasts from the explanted biopsies and the original scars were grown in cell culture, and cell proliferation studies were performed and the results compared. There was a dose response to TGF-beta2, with 200 ng showing the greatest effect. From the original scar specimens, keloid scars demonstrated the greatest cell proliferation kinetics--significantly faster than nonburn and burn hypertrophic scars. After treatment with TGF-beta2, both keloids and burn hypertrophic scars showed an increase in their cell proliferation kinetics compared with vehicle alone. This was not demonstrated with the nonburn hypertrophic scars. Elevated levels of TGF-beta2 are a major contributing factor to the process of proliferative scars, but because nonburn hypertrophic scars do not result in an equally increased response to this cytokine, a truly causative role for this cytokine cannot be promulgated. Rather, it is the combination of the proliferative scar fibroblasts' abnormal response to TGF-beta2 stimulation and elevated levels of this cytokine that controls more accurately the process of keloid and burn hypertrophic scar formation. PMID- 10454328 TI - Reverse venous outflow of a free fibular osteocutaneous flap: a salvage procedure. AB - The authors report 2 patients with a massive bony defect of the tibia due to chronic osteomyelitis. They reconstructed the defect using a free vascularized fibular osteocutaneous flap. Unfortunately, venous insufficiency was diagnosed 24 hours postoperatively. The previous anastomosed veins were promptly explored. The peroneal veins of the vascularized fibular bone graft were noted to be full of thrombi. After thrombectomy, the vessels became very fragile and broke down easily. It was impossible to achieve normal antegrade venous outflow from the previous vein of the donor graft; however, they found that distal runoff of the peroneal vein achieved a reverse venous outflow from the donor graft. The great saphenous vein was dissected and reanastomosed to achieve adequate venous drainage. This procedure may offer an alternative treatment for a flap with venous insufficiency. PMID- 10454329 TI - The use of "excess" soft tissue in the repair of median facial clefts: a report of two cases. AB - The authors present an unusual case of median facial cleft in an infant with prolabiumlike soft-tissue excess between the cleft elements. This tissue was used to augment the nasal tip and lip tubercle. In another patient with median cleft, the authors were conservative in excising the midline structures during the repair, with beneficial effects. PMID- 10454330 TI - Dynamic reconstruction of the abdominal wall using a reinnervated free rectus femoris muscle transfer. AB - Dynamic reconstruction of the abdominal wall using a free reinnervated rectus femoris muscle and an island tensor fascia lata transfer was performed for a large herniation with loss of the bilateral rectus abdominis muscles of the abdominal wall. The tensor fascia lata transfer was used to close an inner side of the abdominal defect, and the rectus femoris muscle replaced the rectus abdominis muscle deficit. The motor nerve of the rectus femoris muscle was sutured to the motor branch of the intercostal nerve. Postoperatively, the transferred rectus femoris muscle was reinnervated via electromyography, and there was no abdominal protrusion and no hernia recurrence. PMID- 10454331 TI - Symptomatic forearm muscle hernia: repair by autologous fascia lata inlay. AB - Eleven cases of symptomatic muscle hernias of the forearm requiring surgical intervention have been described previously. Pain on extremity exertion and an unaesthetic bulge of the forearm were the primary indications for surgery. Advocated treatment modalities range from forearm fasciotomy to anatomic repair of the fascial defect. Although fasciotomy relieves the narrow fascial constriction around the herniated muscle reliably, it often yields an unappealing forearm deformity and incomplete resolution of pain on extremity exertion. Anatomic repair provides the theoretical advantage of restoring normal muscle fascia relationships while concomitantly improving the aesthetic appearance of the extremity. The authors report a case of symptomatic forearm muscle herniation treated successfully with an autologous fascia lata inlay graft. PMID- 10454332 TI - Quadrangular space syndrome associated with superficial radial sensory neuropathy. AB - Compression of the axillary nerve in the quadrangular space is an unusual cause of pain and paresthesia of the upper extremity. In this report, the authors present a patient with a 1-year history of an undiagnosed axillary nerve compression associated with radial sensory neuropathy who improved after surgical decompression of the quadrangular space. PMID- 10454333 TI - Epidermal nevus syndrome: a review and case report. AB - The epidermal nevus syndrome is a disease complex of epidermal nevi and developmental abnormalities of different organ systems. The authors present a case of congenital systematized epidermal nevus syndrome in a patient with skin lesions covering approximately 80% of the total body surface area. The patient underwent staged treatment of the epidermal nevi with a carbon dioxide laser utilizing two different techniques. The larger verrucous lesions were debulked initially, and the resulting defects and thinner lesions were treated using the Silk Touch modality. The lesions were dermaplaned sequentially until they were ablated completely. All wounds healed without complication, and in a 2-year follow-up the patient has experienced no recurrence in the treated areas. Epidermal nevi can be treated safely, effectively, and without recurrence with carbon dioxide laser ablation. PMID- 10454334 TI - Clear-cell basal cell carcinoma of the upper eyelid: a case report. AB - A clear-cell variant of basal cell carcinoma in a previously unreported site is described. This rare variant appeared on the upper eyelid of an 81-year-old woman. Light microscopic examination showed an intradermal tumor with circumscribed lobules composed of clear cells. Histological differential diagnosis of tumors with clear-cell formation is often confusing and can have a great influence on treatment decision making, including surgical intervention. PMID- 10454335 TI - Re: Striae distensae of augmented breasts after oral contraceptives. PMID- 10454336 TI - Massive abdominal wall hernia--coincidence of bilateral semilunar hernias and a linea alba hernia. PMID- 10454337 TI - Re: The sliding door flap for repair of vermilion defects. PMID- 10454338 TI - Multiple trichoepithelioma with basal cell carcinoma. PMID- 10454339 TI - Regulation of tumor growth and metastasis by interleukin-10: the melanoma experience. AB - Because interleukin-10 (IL-10) has potent immunosuppressive and anti-inflammatory properties and is produced by some cancers, including melanoma, we hypothesized that its production by tumor cells may contribute to the escape from immune surveillance. To test this hypothesis, we transfected human A375P melanoma cells that do not express IL-10 with the murine IL-10 gene and subsequently analyzed for changes in tumor growth and metastasis in nude mice. Surprisingly, IL-10 gene transfer resulted in a loss of metastasis and significant inhibition of tumor growth. In addition, the growth of other murine or human melanoma cells was also inhibited when they were admixed with IL-10-transfected cells before injection into nude mice. We provide evidence that IL-10 exerts its antitumor and antimetastatic activity by inhibiting angiogenesis in vivo. The in vivo decrease in neovascularization found in IL-10-secreting tumors is most likely due to the ability of IL-10 to downregulate the synthesis of vascular endothelial growth factor (VEGF), interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF alpha), IL-6, and matrix metalloproteinase-9 (MMP-9) in tumor-associated macrophages. Other studies have shown that IL-10 inhibits tumor metastasis through a natural killer (NK) cell-dependent mechanism. The inhibitory effects of IL-10 on tumor growth and metastasis were also demonstrated in other tumor models, including breast cancers. Furthermore, administration of rIL-10 into mice resulted in inhibition of tumor metastasis. Because IL-10 has little toxicity when given systemically to human volunteers, its efficacy as an antimetastatic agent should be further explored, both as an independent and in combination with other inhibitors of neovascularization. PMID- 10454340 TI - Effectiveness of IFN-gamma for liver abscesses in chronic granulomatous disease. AB - In chronic granulomatous disease, interferon-gamma (IFN-gamma) significantly reduces the incidence and severity of recurrent infections, but its effectiveness administered ex novo during acute infection has been reported in only one case. In this report, we describe two adult brothers with chronic granulomatous disease treated successfully with IFN-gamma for acute liver abscesses. Two brothers with severe recurrent infections of unknown origin were hospitalized for septic fever, malnutrition, and ultrasonographic evidence of liver abscess. Autosomal recessive chronic granulomatous disease was diagnosed based on lack of superoxide anion production by phagocytes and absence of p47-phox protein. An antibiotic regimen specifically directed against Staphylococcus aureus was ineffective, whereas treatment with 50 microg/m2 IFN-gamma s.c. thrice weekly induced complete healing with scarring within 3 months. No septic recurrence was observed during a 4-year follow-up period. In chronic granulomatous disease, IFN-gamma is effective not only in preventing but also in healing life-threatening acute infections. PMID- 10454341 TI - Cooperation of the transcriptional coactivators CBP and p300 with Stat6. AB - Interleukin-4 (IL-4) functions as a critical regulatory cytokine of the immune response. A major effect of IL-4 is the induction of specific gene expression mediated by activation of a latent transcription factor, Stat6. To understand the mechanism by which Stat6 induces gene transcription, the effects of two histone acetylase coactivators, CREB binding protein (CBP) and p300, were evaluated. Both CBP and p300 were found to cooperate with Stat6 for induction of Stat6-dependent transcription. This cooperation does not appear to be due to acetylation of Stat6. The adenoviral E1A oncoprotein, known to bind CBP and p300, can inhibit the ability of CBP and p300 to function as coactivators of Stat6. The cooperative effect of CBP and p300 depends on the presence of a carboxyl-terminal region of Stat6. Stat6 molecules lacking this region behave as negative interfering molecules for Stat6-dependent transcription. Point mutations within this region also affect transcription by Stat6 in response to IL-4, identifying a motif that appears to be required for transcription, possibly through functional cooperation with CBP/p300. PMID- 10454343 TI - Tumor necrosis factor-alpha signals to the IFN-gamma receptor complex to increase Stat1alpha activation. AB - We describe a novel mechanism of signaling interaction through which tumor necrosis factor-alpha (TNF-alpha) treatment augments interferon-gamma (IFN-gamma) induced Stat1alpha DNA-binding complexes and transcriptional activation of a Stat binding element. In TNF-alpha-treated cells, IFN-gamma-induced phosphorylation of Jak2 kinase is increased, Jak2 kinase activity is enhanced, and genetic studies indicate that TNF-alpha requires Jak2 kinase activity to enhance Stat1alpha tyrosine phosphorylation. Increased Jak2 and Stat1alpha phosphorylation are observed within minutes of coexposure to TNF-alpha/IFN-gamma, suggesting a direct signaling interaction. IFN-gamma receptor chain 1 (IFNGR-1) tyrosine phosphorylation is markedly enhanced in cells treated with TNF-alpha/IFN-gamma without alteration in receptor levels. Thus, there exists a direct signaling interaction between TNF-alpha and IFN-gamma, independent of cooperating enhancer elements, that may be relevant for cytokine action during immune-mediated host defense and inflammatory processes. PMID- 10454342 TI - Enhancing in vivo tumorigenicity of B16 melanoma by overexpressing interferon regulatory factor-2: resistance to endogenous IFN-gamma. AB - We investigated the role of interferon (IFN) regulatory factor-2 (IRF-2) as an oncoprotein in vivo, opposing endogenous IFN-gamma suppression of tumor growth. Using syngeneic IFN-gamma knockout mice, we show that endogenous IFN-gamma slows growth of the mouse melanoma cell line B16-F10 in immunocompetent mice, suggesting that tumor cell resistance to IFN-gamma may lead to greater tumorigenicity. IRF-2 is a nuclear transcription factor induced by IFN-gamma that represses numerous IFN-inducible genes, including genes that regulate cell growth, in opposition to the transcriptional activator IRF-1. B16-F10 has a marked growth inhibitory response to IFN-gamma in vitro and has very little IRF-2 induction compared with other murine tumor cell lines. We engineered B16-F10 cells to stably overexpress murine IRF-2. In vitro, these transfected cells showed a marked resistance to the growth-inhibitory effect of IFN-gamma. In normal mice the IRF-2-transfected cells grew much faster than control tumors. In syngeneic IFN-gamma knockout mice, control cells grew at a rate similar to that of IRF-2-transfected cells, implicating resistance to endogenous IFN-gamma as playing the major role in enhanced growth of IRF-2-transfected tumors in intact mice. These experiments demonstrate that (1) IRF-2 enhances B16 melanoma growth and increases resistance to IFN-gamma in vitro, and (2) IRF-2 opposes the growth suppression mediated by endogenous IFN-gamma in vivo. PMID- 10454344 TI - Comparative effects of interleukin-12 and interleukin-4 on cytokine responses by antigen-stimulated memory CD4+ T cells of cattle: IL-12 enhances IFN-gamma production, whereas IL-4 has marginal effects on cytokine expression. AB - Interleukin-12 (IL-12) and IL-4 are important immunoregulatory cytokines that determine the fate of naive T cells during antigen priming in mice and also influence cytokine synthesis by differentiated murine and human T cells. The roles of these cytokines in regulating the differentiation and effector function of bovine T cells are less well studied. We investigated the ability of human IL 12 and bovine IL-4 to modify cytokine expression by antigen-stimulated T cells from cattle immune to the protozoal parasites Babesia bovis and Babesia bigemina or reactive with Mycobacterium bovis purified protein derivative. Peripheral blood mononuclear cells (PBMC) were cultured with specific antigen and IL-4 or IL 12 for 1 week. Then viable lymphoblasts consisting of predominantly CD4+ T cells were restimulated with antigen and antigen-presenting cells (APC) with or without cytokine. Cell lines were cultured for several weeks, and following restimulation with antigen and APC in the absence of exogenous cytokine, the cell lines were analyzed for proliferation, interferon-gamma (IFN-gamma) production, and expression of IL-2, IL4-, IL-10, or IFN-gamma transcript levels using a quantitative competitive RT-PCR. IL-12 and IL-4 had no effect on the composition of CD4, CD8, or gammadelta T cells in the cell lines or on the level of antigen induced proliferation. IL-12 stimulated enhanced levels of IFN-gamma protein and transcript expression in all cell lines, with no consistent effects on IL-2 or IL 4 expression. In two B. bovis-specific cell lines, IL-12 suppressed IL-10 expression. IL-4 had no consistent effect on expression of any cytokine. These results indicate the use of IL-12 as an adjuvant to enhance type 1 cytokine responses in cattle during antigen priming. PMID- 10454345 TI - Production of influenza-stimulated tumor necrosis factor-alpha by monocytes following acute influenza infection in humans. AB - Tumor necrosis factor-alpha (TNF-alpha) is often measured in the serum or plasma of patients with severe infections, and marked elevation correlates with poor outcome. The relationship of TNF-alpha to protection from disease is frequently not observed because prospective studies of infectious agents are difficult to perform. We took advantage of a human antiviral influenza challenge study to correlate TNF-alpha production with seroconversion and symptom development. TNF alpha production was measured by ELISA in the plasma compartment or was measured by intracellular production at the single cell level in the monocyte gated population. Monocyte TNF-alpha was associated with asymptomatic seroconversion, whereas there was no change in the plasma at the times measured. Measurement of TNF-alpha at the single cell level by flow cytometry may allow for better differentiation of the protective role of this cytokine in future studies. PMID- 10454346 TI - Elevated serum thrombopoietin and interleukin-6 concentrations in thrombocytosis associated with inflammatory bowel disease. AB - Reactive thrombocytosis is a typical feature in inflammatory bowel disease (IBD). The question arose as to whether the normal negative feedback regulation of the concentration of thrombopoietin (TPO) in blood was altered in IBD patients. We measured serum immunoreactive TPO in 30 patients with active IBD, 29 patients with inactive IBD, and 56 healthy controls. The results were related to platelet and leukocyte counts and to the serum concentration of interleukin 6 (IL-6). Patients with active IBD exhibited significantly increased TPO levels (medians 112 pg/ml vs. 90 pg/ml in controls, p < 0.05) in association with thrombocytosis (428 platelets/nl blood vs. 241 platelets/nl blood in controls), leukocytosis, and increased IL-6 levels (12.9 pg/ml vs. 2.5 pg/nl in controls). In patients with inactive IBD, only platelets (322/nl) and leukocytes were above normal. Although the observation of increased TPO and IL-6 levels provides an explanation for the occurrence of thrombocytosis in IBD, the pathogenetic mechanisms underlying the elevated TPO level still need to be identified. PMID- 10454347 TI - Modulation of monocyte chemokine production and nuclear factor kappa B activity by oxidants. AB - Reactive oxygen species can directly damage tissue. In this setting, amplification of tissue damage also occurs through infiltration of inflammatory cells either acutely or chronically. Several recent studies suggest that reactive oxygen species stimulate production of certain chemokines, which are potent chemoattractants for inflammatory cells. In the present study, we examined whether oxidants, generated by the combination of xanthine and xanthine oxidase (X/XO), alter chemokine production by monocytes and U937 cells. Our findings demonstrate that X/XO stimulates monocytes, but not U937 cells, to produce increased amounts of interleukin-8 (IL-8) and monocyte chemoattractant protein. This effect is attenuated by pretreatment with dimethylsulfoxide (DMSO), a scavenger of hydroxyl radicals, but is not affected by superoxide dismutase or catalase. In contrast, X/XO-induced cytotoxicity, evidenced by lactate dehydrogenase release, is mediated primarily by hydrogen peroxide, as catalase reverses this effect. Finally, exposure to X/XO causes an increase in nuclear factor kappa B (NF-kappaB), and this effect is attenuated by DMSO. These studies suggest that reactive oxygen species can induce production of molecules that amplify inflammation through attraction of inflammatory cells. It appears the hydroxyl radical is the principal oxidant species involved in stimulation of chemokine production. PMID- 10454348 TI - Intracellular analysis of interleukin-2 induction provides direct evidence at the single cell level of differential coactivation requirements for CD45RA+ and CD45RO+ T cell subsets. AB - Through measurements of intracellular cytokine production, evidence is provided at the single cell level that triggering different cell surface molecules preferentially activates discrete human peripheral blood (PB) T cell subsets. T cell costimulation due to cross-linking a variety of individual molecules (beta1, beta2, and beta7 integrins, CD26, CD43, or CD44), in conjunction with the CD3/TCR complex, preferentially activated CD45RO+ PB T lymphocytes. CD28, however, costimulated interleukin-2 (IL-2) production in both CD45RO+ and CD45RA+ subpopulations. The amount of soluble IL-2 produced by CD28 coactivation was 15 30-fold higher than that due to integrin or CD26-dependent coactivation, although even the lowest amount of soluble IL-2 produced was in the range of the high affinity IL-2 receptor. The overall proliferative responses were similar among all costimulatory settings. This was in part due to the uniform upregulation of IL-2 receptor-alpha (IL-2R alpha) (CD25) expression on the entire T cell population activated under each of the different costimulatory conditions. The data provide direct evidence on a single cell level that activation of human CD45RA+ (naive) T cells is stringently controlled and, in these studies, limited to CD28 costimulation for induction of IL-2 production. In contrast, coactivation of CD45RO+ (memory) T lymphocytes can proceed by a variety of different PB T cell surface molecules. PMID- 10454349 TI - Detection and characterization of antibodies to PEG-IFN-alpha2b using surface plasmon resonance. AB - Some patients treated with type I interferon (IFN) preparations develop neutralizing antibodies that may abrogate any clinical benefit. We have a new complex of polyethylene glycol12000 and IFN-alpha2b (PEG-IFN-alpha2b) in clinical trials and need to be able to detect any antibodies formed specifically against the complex. We have, therefore, devised a method based on measurement of surface plasmon resonance (SPR) in the BIACORE 2000 apparatus. PEG-IFN-alpha2b is anchored to one flow cell on the sensor chip, IFN-alpha2b to another, and PEG to a third. A 20 microl serum sample flows in turn through the three cells, which are optically scanned. Any antibodies in the serum bind to the corresponding immobilized antigen, and a change in the optical signal is generated. With appropriate specific reagents, their immunoglobulin isotype can be similarly established. The automated assay can quickly test numerous sera. Very little serum is needed, and the assay is reliable and precise and can detect low alphaffinity antibodies. PMID- 10454350 TI - Antioxidants attenuate acute toxicity of tumor necrosis factor-alpha induced by brain injury in rat. AB - Tumor necrosis factor-alpha alpha (TNF-alpha) and reactive oxygen species (ROS) are produced in the brain after traumatic injury and have deleterious effects. In a rat model of closed head injury (CHI), the synthetic antioxidant from the nitroxide family, Tempol, improved recovery and protected the blood-brain barrier. Similar protection was found after CHI in heat-acclimated rats, in which the endogenous antioxidants have been shown to be elevated after CHI. The present study examined the relationship between TNF-alpha and ROS after CHI, namely, whether after CHI, antioxidants that afforded cerebroprotection also attenuated brain levels of TNF-alpha. Three groups of rats were subjected to CHI: (1) control, nontreated, (2) Tempol-treated, and (3) heat-acclimated (30 days at 34 degrees C). Four hours after injury (time for peak production of TNF-alpha), the activity of TNF-alpha was measured. Although clinical recovery was facilitated in rats of the two treated groups, TNF-alpha activity was as high as in the traumatized, untreated rats. Moreover, direct injection of TNF-alpha into mouse brain induced disruption of the blood-brain barrier, indicating its acute harmful effect. This toxic effect was attenuated by before and after treatment with Tempol. Our results support the hypothesis that in vivo antioxidants neutralize TNF-alpha toxicity, probably by interfering with activation of the transcription factor NF-kappa-B. PMID- 10454351 TI - Activation of the Jak-Stat pathway in cells that exhibit selective sensitivity to the antiviral effects of IFN-beta compared with IFN-alpha. AB - We determined whether selective activation of components of the Jak-Stat pathway by different type I interferons (IFN) occurs in human myocardial fibroblasts that exhibit much higher sensitivity to the antiviral effects of IFN-beta than of IFN alpha. Similar levels of activation of the Tyk2 kinase and the Stat3 transcription factor were induced in response to either IFN-beta or IFN-alpha treatment. However, activation of the Jak1 tyrosine kinase was detectable only in IFN-beta-treated but not IFN-alpha-treated cells. Consistent with this, tyrosine phosphorylation of Stat1 and Stat2 and formation of the IFN-stimulated gene factor 3 (ISGF3) complex occurred to a much higher degree in response to IFN-beta stimulation. These findings demonstrate that differential activation of distinct components of the Jak-Stat pathway by different type I IFN can occur. Furthermore, they strongly suggest that such selective activation accounts for the occurrence of differences in the antiviral properties of distinct type I IFN in certain cell types. PMID- 10454352 TI - Role of IFN-gamma produced after intraperitoneal transplantation of AK-5 cells in the induction of Fas ligand expression by tumor cells leading to immune evasion. AB - AK-5 tumor cells expressed Fas-L on their surface after intraperitoneal transplantation in syngeneic animals. Fas-L expression by AK-5 cells is involved in the killing of the effector cells. Thus, the tumor has developed an escape mechanism from immune attack. In the present study, we showed that Fas-L expression on AK-5 cells is regulated by interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), as injection of antibodies against IFN-gamma downregulated the expression of Fas-L by tumor cells as determined by immunostaining and Northern hybridizations. Fas-L present on the tumor cells is biologically functional, as it induced DNA fragmentation in Fas+ YAC-1 cells. We have also shown shedding of Fas-L in cell-free ascitic fluid from tumor-bearing animals. These observations suggest that such cytokines as IFN-gamma and TNF alpha play an important role in regulating the expression of Fas-L by AK-5 cells. PMID- 10454353 TI - Reduced K+ channel inactivation, spike broadening, and after-hyperpolarization in Kvbeta1.1-deficient mice with impaired learning. AB - A-type K+ channels are known to regulate neuronal firing, but their role in repetitive firing and learning in mammals is not well characterized. To determine the contribution of the auxiliary K+ channel subunit Kvbeta1.1 to A-type K+ currents and to study the physiological role of A-type K+ channels in repetitive firing and learning, we deleted the Kvbeta1.1 gene in mice. The loss of Kvbeta1.1 resulted in a reduced K+ current inactivation in hippocampal CA1 pyramidal neurons. Furthermore, in the mutant neurons, frequency-dependent spike broadening and the slow afterhyperpolarization (sAHP) were reduced. This suggests that Kvbeta1.1-dependent A-type K+ channels contribute to frequency-dependent spike broadening and may regulate the sAHP by controlling Ca2+ influx during action potentials. The Kvbeta1.1-deficient mice showed normal synaptic plasticity but were impaired in the learning of a water maze test and in the social transmission of food preference task, indicating that the Kvbeta1.1 subunit contributes to certain types of learning and memory. PMID- 10454354 TI - Deficits in memory tasks of mice with CREB mutations depend on gene dosage. AB - Studies in Aplysia, Drosophila, and mice have shown that the transcription factor CREB is involved in formation and retention of long-term memory. To analyze the impact of differential CREB levels on learning and memory, we varied the gene dosage of CREB in two strains of mutant mice: (1) CREBalphadelta mice, in which the alpha and delta isoforms are disrupted, but a third isoform beta is strongly up-regulated; (2) CREBcomp, a compound strain with one alphadelta allele and one CREBnull allele in which all CREB isoforms are disrupted. To minimize genetic background effects, CREB mutations were backcrossed into a C57BL/6 and a FVB/N strain, respectively, and studies were performed in F1 hybrids from these lines. CREBcomp but not CREBalphadelta F1 hybrids were impaired in water maze learning and fear conditioning, demonstrating a CREB gene dosage effect. However, analysis of the platform searching strategies in the water maze task suggested that CREBcomp mutants are impaired in behavioral flexibility rather than in spatial memory. In contrast to previous experiments using CREBalphadelta mice with different genetic background, the F1 hybrid CREBalphadelta and CREBcomp mice did not show deficits in a social transmission of food preference task nor in dentate gyrus and CA1 LTP as recorded from slice preparations. These data indicate that the hybrid vigor typical for F1 hybrids may compensate for a reduction in CREB levels in some tests. On the other hand, the persistence of clear behavioral deficits as shown by the F1 hybrid CREBcomp mice in water maze and fear conditioning indicates a robust and repeatable phenomenon that will permit further functional analysis of CREB. PMID- 10454355 TI - Hyperactivity and learning deficits in transgenic mice bearing a human mutant thyroid hormone beta1 receptor gene. AB - Resistance to thyroid hormone (RTH) is a human syndrome mapped to the thyroid receptor beta (TRbeta) gene on chromosome 3, representing a mutation of the ligand-binding domain of the TRbeta gene. The syndrome is characterized by reduced tissue responsiveness to thyroid hormone and elevated serum levels of thyroid hormones. A common behavioral phenotype associated with RTH is attention deficit hyperactivity disorder (ADHD). To test the hypothesis that RTH produces attention deficits and/or hyperactivity, transgenic mice expressing a mutant TRbeta gene were generated. The present experiment tested RTH transgenic mice from the PV kindred on behavioral tasks relevant to the primary features of ADHD: hyperactivity, sustained attention (vigilance), learning, and impulsivity. Male transgenic mice showed elevated locomotor activity in an open field compared to male wild-type littermate controls. Both male and female transgenic mice exhibited impaired learning of an autoshaping task, compared to wild-type controls. On a vigilance task in an operant chamber, there were no differences between transgenics and controls on the proportion of hits, response latency, or duration of stimulus tolerated. On an operant go/no-go task measuring sustained attention and impulsivity, there were no differences between controls and transgenics. These results indicate that transgenic mice bearing a mutant human TRbeta gene demonstrate several behavioral characteristics of ADHD and may serve a valuable heuristic role in elucidating possible candidate genes in converging pathways for other causes of ADHD. PMID- 10454356 TI - Alpha7 nicotinic receptor subunits are not necessary for hippocampal-dependent learning or sensorimotor gating: a behavioral characterization of Acra7-deficient mice. AB - The alpha7 nicotinic acetylcholine receptor (nAChR) subunit is abundantly expressed in the hippocampus and contributes to hippocampal cholinergic synaptic transmission suggesting that it may contribute to learning and memory. There is also evidence for an association between levels of alpha7 nAChR and in sensorimotor gating impairments. To examine the role of alpha7 nAChRs in learning and memory and sensorimotor gating, Acra7 homozygous mutant mice and their wild type littermates were tested in a Pavlovian conditioned fear test, for spatial learning in the Morris water task, and in the prepulse inhibition paradigm. Exploratory activity, motor coordination, and startle habituation were also evaluated. Acra7 mutant mice displayed the same levels of contextual and auditory cue condition fear as wild-type mice. Similarly, there were no differences in spatial learning performance between mutant and wild-type mice. Finally, Acra7 mutant and wild-type mice displayed similar levels of prepulse inhibition. Other behavioral responses in Acra7 mutant mice were also normal, except for an anxiety related behavior in the open-field test. The results of this study show that the absence of alpha7 nAChRs has little impact on normal, base-line behavioral responses. Future studies will examine the contribution of alpha7 nAChR to the enhancement of learning and sensorimotor gating following nicotine treatments. PMID- 10454357 TI - Enhanced hippocampal CA1 LTP but normal spatial learning in inositol 1,4,5 trisphosphate 3-kinase(A)-deficient mice. AB - To define the physiological role of IP(3)3-kinase(A) in vivo, we have generated a mouse strain with a null mutation of the IP(3)3-kinase(A) locus by gene targeting. Homozygous mutant mice were fully viable, fertile, apparently normal, and did not show any morphological anomaly in brain sections. In the mutant brain, the IP4 level was significantly decreased whereas the IP3 level did not change, demonstrating a major role of IP(3)3-kinase(A) in the generation of IP4. Nevertheless, no significant difference was detected in the hippocampal neuronal cells of the wild-type and the mutant mice in the kinetics of Ca2+ regulation after glutamate stimulation. Electrophysiological analyses carried out in hippocampal slices showed that the mutation significantly enhanced the LTP in the hippocampal CA1 region, but had no effect on the LTP in dentate gyrus (DG). No difference was noted, however, between the mutant and the wild-type mice in the Morris water maze task. Our results indicate that IP(3)3-kinase(A) may play an important role in the regulation of LTP in hippocampal CA1 region through the generation of IP4, but the enhanced LTP in the hippocampal CA1 does not affect spatial learning and memory. PMID- 10454358 TI - Selective abolition of the NMDA component of long-term potentiation in mice lacking mGluR5. AB - The mechanisms underlying the differential expression of long-term potentiation (LTP) by AMPA and NMDA receptors, are unknown, but could involve G-protein-linked metabotropic glutamate receptors. To investigate this hypothesis we created mutant mice that expressed no metabotropic glutamate receptor 5 (mGluR5), but showed normal development. In an earlier study of these mice we analyzed field excitatory postsynaptic potential (fEPSPs) in CA1 region of the hippocampus and found a small decrease; possibly arising from changes in the NMDAR-mediated component of synaptic transmission. In the present study we used whole-cell patch clamp recordings of evoked excitatory postsynaptic currents (EPSCs) in CA1 pyramidal neurons to identify the AMPAR- and NMDAR-mediated components of LTP. Recordings from control mice following tetanus, or agonist application (IS, 3R-1 amino-cyclopentane 1,3-dicarboxylic acid) (ACPD), revealed equal enhancement of the AMPA and NMDA receptor-mediated components. In contrast, CA1 neurons from mGluR5-deficient mice showed a complete loss of the NMDA-receptor-mediated component of LTP (LTP(NMDA)), but normal LTP of the AMPA-receptor-mediated component (LTP(AMPA)). This selective loss of LTP(NMDA) was seen in three different genotypic backgrounds and was apparent at all holding potentials (-70 mV to +20 mV). Furthermore, the LTP(NMDA) deficit in mGluR5 mutant mice could be rescued by stimulating protein kinase C (PKC) with 4beta-phorbol-12,13-dibutyrate (PDBu). These results suggest that PKC may couple the postsynaptic mGluR5 to the NMDA-receptor potentiation during LTP, and that this signaling mechanism is distinct from LTP(AMPA). Differential enhancement of AMPAR and NMDA receptors by mGluR5 also supports a postsynaptic locus for LTP. PMID- 10454359 TI - CA1 long-term potentiation is diminished but present in hippocampal slices from alpha-CaMKII mutant mice. AB - Previous work has shown that mice missing the alpha-isoform of calcium-calmodulin dependent protein kinase II (alpha-CaMKII) have a deficiency in CA1 hippocampal long-term potentiation (LTP). Follow-up studies on subsequent generations of these mutant mice in a novel inbred background by our laboratories have shown that whereas a deficiency in CA1 LTP is still present in alpha-CaMKII mutant mice, it is different both quantitatively and qualitatively from the deficiency first described. Mice of a mixed 129SvOla/SvJ;BALB/c;C57B1/6 background derived from brother/sister mating of the alpha-CaMKII mutant line through multiple generations (>10) were produced by use of in vitro fertilization. Although LTP at 60 min post-tetanus was clearly deficient in these (-/-) alpha-CaMKII mice (42.6%, n = 33) compared with (+/+) alpha-CaMKII control animals (81.7%, n = 17), alpha-CaMKII mutant mice did show a significant level of LTP. The amount of LTP observed in alpha-CaMKII mutants was normally distributed, blocked by APV (2.7%, n = 8), and did not correlate with age. Although this supports a role for alpha CaMKII in CA1 LTP, it also suggests that a form of alpha-CaMKII-independent LTP is present in mice that could be dependent on another kinase, such as the beta isoform of CaMKII. A significant difference in input/output curves was also observed between (-/-) alpha-CaMKII and (+/+) alpha-CaMKII animals, suggesting that differences in synaptic transmission may be contributing to the LTP deficit in mutant mice. However, tetani of increasing frequency (50, 100, and 200 Hz) did not reveal a higher threshold for potentiation in (-/-) alpha-CaMKII mice compared with (+/+) alpha-CaMKII controls. PMID- 10454361 TI - Different training procedures recruit either one or two critical periods for contextual memory consolidation, each of which requires protein synthesis and PKA. AB - We have used a combined genetic and pharmacological approach to define the time course of the requirement for protein kinase A (PKA) and protein synthesis in long-term memory for contextual fear conditioning in mice. The time course of amnesia in transgenic mice that express R(AB) and have genetically reduced PKA activity in the hippocampus parallels that observed both in mice treated with inhibitors of PKA and mice treated with inhibitors of protein synthesis. This PKA and protein synthesis-dependent memory develops between 1 hr and 3 hr after training. By injecting the protein synthesis inhibitor anisomycin or the PKA inhibitor Rp-cAMPs at various times after training, we find that depending on the nature of training, contextual memory has either one or two brief consolidation periods requiring synthesis of new proteins, and each of these also requires PKA. Weak training shows two time periods of sensitivity to inhibitors of protein synthesis and PKA, whereas stronger training exhibits only one. These studies underscore the parallel dependence of long-term contextual memory on protein synthesis and PKA and suggest that different training protocols may recruit a common signaling pathway in distinct ways. PMID- 10454360 TI - Impaired eye-blink conditioning in waggler, a mutant mouse with cerebellar BDNF deficiency. AB - In addition to their trophic functions, neurotrophins are also implicated in synaptic modulation and learning and memory. Although gene knockout techniques have been used widely in studying the roles of neurotrophins at molecular and cellular levels, behavioral studies using neurotrophin knockouts are limited by the early-onset lethality and various sensory deficits associated with the gene knockout mice. In the present study, we found that in a spontaneous mutant mouse, waggler, the expression of brain-derived neurotrophic factor (BDNF) was selectively absent in the cerebellar granule cells. The cytoarchitecture of the waggler cerebellum appeared to be normal at the light microscope level. The mutant mice exhibited no sensory deficits to auditory stimuli or heat-induced pain. However, they were massively impaired in classic eye-blink conditioning. These results suggest that BDNF may have a role in normal cerebellar neuronal function, which, in turn, is essential for classic eye-blink conditioning. PMID- 10454363 TI - Automated measurement of mouse freezing behavior and its use for quantitative trait locus analysis of contextual fear conditioning in (BALB/cJ x C57BL/6J)F2 mice. AB - The most commonly measured mouse behavior in fear conditioning tests is freezing. A technical limitation, particularly for genetic studies, is the method of direct observation used for quantifying this response, with the potential for bias or inconsistencies. We report the use of a computerized method based on latency between photobeam interruption measures as a reliable scoring criterion in mice. The different computer measures obtained during contextual fear conditioning tests showed high correlations with hand-scored freezing; r values ranged from 0.87 to 0.94. Previously reported strain differences between C57BL/6J and DBA/2J in context-dependent fear conditioning were also detected by the computer-based system. In addition, the use of computer-scored freezing of 199 (BALB/cJ x C57BL/6J)F2 mice enabled us to detect a suggestive gender-dependent chromosomal locus for contextual fear conditioning on distal chromosome 8 by QTL analysis. Automation of freeze scoring would significantly increase efficiency and reliability of this learning and memory test. PMID- 10454362 TI - Increase in syntaxin 1B mRNA in hippocampal and cortical circuits during spatial learning reflects a mechanism of trans-synaptic plasticity involved in establishing a memory trace. AB - It has long been proposed that the cellular and molecular mechanisms responsible for LTP may well involve the mechanisms that lead to the type of synaptic modification that occurs during learning. However, it is also known that a single memory trace is encoded in spatially distributed networks; implying that alterations of synaptic strength occur at multiple sites along circuits of connected cells. Recent evidence suggests that regulation of the gene encoding syntaxin 1B, a presynaptic protein involved in exocytosis, plays an important role in the mediation of trans-synaptic LTP, a candidate mechanism for the propagation of plasticity in neural circuits during learning. Using in situ hybridization to measure the mRNA levels at different time points after learning a spatial working or reference memory task, we show that expression of the gene encoding this protein in the hippocampal and corticoprefrontal circuits increases linearly with performance at a critical window of learning when rats are reaching between 75% and 100% of their maximal performance. No changes were observed during the early phases of learning or when rats where overtrained. The correlational analysis indicates that coordinated increases in syntaxin 1B expression occurs in hippocampal circuits during working memory and in more widespread hippocampocortical circuits during reference memory. These results suggest that a form of trans-synaptic plasticity mediated in part by regulation of the expression of syntaxin 1B may play an active role in configuring specific spatially distributed circuits during the laying down of memories. PMID- 10454364 TI - Food avoidance learning in squirrel monkeys and common marmosets. AB - Using a conditioned food avoidance learning paradigm, six squirrel monkeys (Saimiri sciureus) and six common marmosets (Callithrix jacchus) were tested for their ability to (1) reliably form associations between visual or olfactory cues of a potential food and its palatability and (2) remember such associations over prolonged periods of time. We found (1) that at the group level both species showed one-trial learning with the visual cues color and shape, whereas only the marmosets were able to do so with the olfactory cue, (2) that all individuals from both species learned to reliably avoid the unpalatable food items within 10 trials, (3) a tendency in both species for quicker acquisition of the association with the visual cues compared with the olfactory cue, (4) a tendency for quicker acquisition and higher reliability of the aversion by the marmosets compared with the squirrel monkeys, and (5) that all individuals from both species were able to reliably remember the significance of the visual cues, color and shape, even after 4 months, whereas only the marmosets showed retention of the significance of the olfactory cues for up to 4 weeks. Furthermore, the results suggest that in both species tested, illness is not a necessary prerequisite for food avoidance learning but that the presumably innate rejection responses toward highly concentrated but nontoxic bitter and sour tastants are sufficient to induce robust learning and retention. PMID- 10454365 TI - Multiple memory processes following training that a food is inedible in Aplysia. AB - In many organisms, memory after training can be separated into a number of processes. We now report that separable memory processes are also initiated by a training procedure affecting Aplysia feeding behavior, a model system for examining the neural mechanisms underlying the regulation of a complex behavior. Four distinct memory process were identified: (1) a very short-term memory that declines within 15 min, (2) a short-term memory that persists for 0.5-1.0 hr, (3) an intermediate-term memory, observed 4 hr after training, and (4) a long-term memory that is seen only after a 12- to 24-hr delay. The four memory processes can be distinguished by the different training procedures that are required to elicit them. A single 5-min training session is sufficient to elicit the very short-term memory. However, a longer training session that continues until the animal stops responding to food is needed to elicit short-term memory. Intermediate-term memory is observed only after a spaced training procedure (three 5-min training sessions separated by 30-min intervals). A single 5-min training session that does not cause either short-term or intermediate-term memory is sufficient to induce long-term memory, indicating that short- and long term memory are independent, parallel processes. Short- and long-term memory can also be separated by the effects of a post-training experience. Long-term, but not short-term, memory can be attenuated by cooling animals immediately after training. Cooling before the training does not affect either the training or the subsequent short- or long-term memory. PMID- 10454366 TI - NMDA receptor antagonism in the lateral/basolateral but not central nucleus of the amygdala prevents the induction of facilitated learning in response to stress. AB - Exposure to an acute stressful event facilitates classical eye-blink conditioning in the male rat. The facilitation persists for days after the stressor and its induction is prevented by antagonism of the N-methyl-D-aspartate (NMDA) type of glutamate receptor. To determine whether NMDA receptor antagonists prevent the facilitated conditioning by activity in the amygdala, a competitive antagonist, AP5, was injected bilaterally into the lateral/basolateral versus central nuclei of the amygdala. Local injection of D,L-2-amino-5-phosphonovaleric acid (AP5) into the lateral/basolateral nucleus before stressor exposure prevented the facilitated learning 24 hr later, whereas antagonism in the central nucleus before stressor exposure did not. To determine when the necessary activation occurs, AP5 was injected into the lateral/basolateral nucleus before versus after exposure to the acute stressful event. Blockade of NMDA receptors before but not after stressor exposure prevented the facilitated acquisition of the conditioning in response to stress. These results suggest that exposure to a stressful event transiently activates NMDA receptors in basolateral/lateral nuclei of the amygdala and thereby induces a persistent enhancement of associative learning. PMID- 10454367 TI - The specific role of cGMP in hippocampal LTP. AB - Previous results have suggested that cGMP is involved in hippocampal long-term potentiation (LTP), perhaps as the presynaptic effector of a retrograde messenger. However, other studies have failed to replicate some of those results, making the role of cGMP uncertain. We therefore reexamined this question and identified several variables that can affect the contribution of cGMP. First, brief perfusion with 8-Br-cGMP before weak tetanic stimulation produced long lasting potentiation in the CA1 region of hippocampal slices, but more prolonged perfusion with 8-Br-cGMP before the tetanus did not produce long-lasting potentiation. Second, the activity-dependent long-lasting potentiation by cGMP analogs was reduced when NMDA receptors were completely blocked, indicating that NMDA receptor activation contributes to, but is not required for, the potentiation. The amount of reduction of the potentiation differed with different protocols, and in some cases could be complete. Third, LTP produced by strong tetanic stimulation in the stratum radiatum of CA1 (which expresses eNOS) was blocked by inhibitors of soluble guanylyl cyclase or cGMP-dependent protein kinase, but LTP in the stratum oriens (which does not express eNOS) was not. The results of these experiments should help to explain some of the discrepant findings from previous studies, and, in addition, may provide insights into the mechanisms and functional role of the cGMP-dependent component of LTP. PMID- 10454368 TI - Differential induction of long-term synaptic facilitation by spaced and massed applications of serotonin at sensory neuron synapses of Aplysia californica. AB - Serotonin (5HT)-induced facilitation of synaptic transmission from tail sensory neurons (SNs) to motor neurons (MNs) in the marine mollusc Aplysia provides a cellular model of short- and long-term memory for behavioral sensitization of the tail withdrawal reflex. Synaptic facilitation at these synapses occurs in three temporal phases: short-term (STF, lasting minutes), intermediate-term (ITF, lasting more than an hour), and long-term (LTF, lasting >24 hr). STF, ITF, and LTF differ in their induction requirements: A single brief exposure of 5HT induces STF, whereas five applications are required for ITF and LTF. Moreover, STF and LTF can be induced independently. Different forms of memory often show differential sensitivity to the pattern of training trials. To begin to explore this effect at a cellular level, we examined ITF and LTF induced by one of two patterns of 5HT application: a spaced pattern (five 5-min exposures with an interval of 15 min) or a massed pattern (one continuous 25-min application). The spaced and massed patterns both induced ITF; however, spaced 5HT application was significantly more reliable at inducing LTF than was massed application. Thus, whereas induction of ITF and LTF require similar amounts of 5HT, the cellular mechanisms underlying the induction of LTF are more sensitive to the pattern of the induction trials. In the massed group, further analysis revealed a relationship between the expression of ITF and the subsequent expression of LTF, suggesting that these two processes may be mechanistically related. PMID- 10454369 TI - What do the mushroom bodies do for the insect brain? an introduction. PMID- 10454370 TI - Evolution, discovery, and interpretations of arthropod mushroom bodies. AB - Mushroom bodies are prominent neuropils found in annelids and in all arthropod groups except crustaceans. First explicitly identified in 1850, the mushroom bodies differ in size and complexity between taxa, as well as between different castes of a single species of social insect. These differences led some early biologists to suggest that the mushroom bodies endow an arthropod with intelligence or the ability to execute voluntary actions, as opposed to innate behaviors. Recent physiological studies and mutant analyses have led to divergent interpretations. One interpretation is that the mushroom bodies conditionally relay to higher protocerebral centers information about sensory stimuli and the context in which they occur. Another interpretation is that they play a central role in learning and memory. Anatomical studies suggest that arthropod mushroom bodies are predominately associated with olfactory pathways except in phylogenetically basal insects. The prominent olfactory input to the mushroom body calyces in more recent insect orders is an acquired character. An overview of the history of research on the mushroom bodies, as well as comparative and evolutionary considerations, provides a conceptual framework for discussing the roles of these neuropils. PMID- 10454371 TI - Tripartite mushroom body architecture revealed by antigenic markers. AB - We have explored the organization of the axonal lobes in Drosophila mushroom bodies by using a panel of immunohistochemical markers. These markers consist of antibodies to eight proteins expressed preferentially in the mushroom bodies: DAMB, DCO, DRK, FASII, LEO, OAMB, PKA RII, and RUT. Previous to this work, four axonal lobes, two projecting dorsally (alpha and alpha') and two medially (beta and gamma), had been described in Drosophila mushroom bodies. However, our analysis of immunohistochemically stained frontal and sagittal sections of the brain revealed three medially projecting lobes. The newly distinguished lobe, which we term beta', lies along the dorsal surface of beta, just posterior to gamma. In addition to resolving a fifth lobe, our studies revealed that there are specific lobe sets defined by equivalent marker expression levels. These sets are (1) the alpha and beta lobes, (2) the alpha' and beta' lobes, and (3) the gamma lobe and heel (a lateral projection formed by a hairpin turn of some of the peduncle fibers). All of the markers we have examined are consistent with these three sets. Previous Golgi studies demonstrate that each mushroom body cell projects one axon that branches into a dorsal lobe and a medial lobe, or one unbranched axon that projects medially. Taken together with the lobe sets listed above, we propose that there are three major projection configurations of mushroom body cell axons: (1) one branch in the alpha and one in the beta lobe, (2) one branch in the alpha' and one in the beta' lobe, and (3) one unbranched axon projecting to the heel and the gamma lobe. The fact that these neuron types exhibit differential expression levels of a number of mushroom body genes suggests that they may have corresponding functional differences. These functions may be conserved in the larvae, as several of these genes were expressed in larval and embryonic mushroom bodies as well. The basic mushroom body structure, including the denritic calyx, peduncle, and lobes, was already visible by the late stages of embryogenesis. With new insights into mushroom body organization, and the characterization of markers for developing mushroom bodies, we are beginning to understand how these structures form and function. PMID- 10454372 TI - The organization of extrinsic neurons and their implications in the functional roles of the mushroom bodies in Drosophila melanogaster Meigen. AB - Although the importance of the Drosophila mushroom body in olfactory learning and memory has been stressed, virtually nothing is known about the brain regions to which it is connected. Using Golgi and GAL4-UAS techniques, we performed the first systematic attempt to reveal the anatomy of its extrinsic neurons. A novel presynaptic reporter construct, UAS-neuronal synaptobrevin-green fluorescent protein (n-syb-GFP), was used to reveal the direction of information in the GAL4 labeled neurons. Our results showed that the main target of the output neurons from the mushroom body lobes is the anterior part of the inferior medial, superior medial, and superior lateral protocerebrum. The lobes also receive afferents from these neuropils. The lack of major output projections directly to the deutocerebrum's premotor pathways discourages the view that the role of the mushroom body may be that of an immediate modifier of behavior. Our data, as well as a critical evaluation of the literature, suggest that the mushroom body may not by itself be a "center" for learning and memory, but that it can equally be considered as a preprocessor of olfactory signals en route to "higher" protocerebral regions. PMID- 10454373 TI - Immunocytochemical mapping of an RDL-like GABA receptor subunit and of GABA in brain structures related to learning and memory in the cricket Acheta domesticus. AB - The distribution of putative RDL-like GABA receptors and of gamma-aminobutyric acid (GABA) in the brain of the adult house cricket Acheta domesticus was studied using specific antisera. Special attention was given to brain structures known to be related to learning and memory. The main immunostaining for the RDL-like GABA receptor was observed in mushroom bodies, in particular the upper part of mushroom body peduncle and the two arms of the posterior calyx. Weaker immunostaining was detected in the distal part of the peduncle and in the alpha and beta lobes. The dorso- and ventrolateral protocerebrum neuropils appeared rich in RDL-like GABA receptors. Staining was also detected in the glomeruli of the antennal lobe, as well as in the ellipsoid body of the central complex. Many neurons clustered in groups exhibit GABA-like immunoreactivity. Tracts that were strongly immunostained innervated both the calyces and the lobes of mushroom bodies. The glomeruli of the antennal lobe, the ellipsoid body, as well as neuropils of the dorso- and ventrolateral protocerebrum were also rich in GABA like immunoreactivity. The data demonstrated a good correlation between the distribution of the GABA-like and of the RDL-like GABA receptor immunoreactivity. The prominent distribution of RDL-like GABA receptor subunits, in particular areas of mushroom bodies and antennal lobes, underlines the importance of inhibitory signals in information processing in these major integrative centers of the insect brain. PMID- 10454374 TI - Early development of mushroom bodies in the brain of the honeybee Apis mellifera as revealed by BrdU incorporation and ablation experiments. AB - In the honeybee the mushroom bodies are prominent neuropil structures arranged as pairs in the dorsal protocerebrum of the brain. Each mushroom body is composed of a medial and a lateral subunit. To understand their development, the proliferation pattern of mushroom body intrinsic cells, the Kenyon cells, were examined during larval and pupal stages using the bromodeoxyuridine (BrdU) technique and chemical ablation with hydroxyurea. By larval stage 1, approximately 40 neuroblasts are located in the periphery of the protocerebrum. Many of these stem cells divide asymmetrically to produce a chain of ganglion mother cells. Kenyon cell precursors underly a different proliferation pattern. With the beginning of larval stage 3, they are arranged in two large distinct cell clusters in each side of the brain. BrdU incorporation into newly synthesized DNA and its immunohistochemical detection show high mitotic activity in these cell clusters that lasts until mid-pupal stages. The uniform diameter of cells, the homogeneous distribution of BrdU-labeled nuclei, and the presence of equally dividing cells in these clusters indicate symmetrical cell divisions of Kenyon cell precursors. Hydroxyurea applied to stage 1 larvae caused the selective ablation of mushroom bodies. Within these animals a variety of defects were observed. In the majority of brains exhibiting mushroom body defects, either one mushroom body subunit on one or on both sides, or three or four subunits (e.g., complete mushroom body ablation) were missing. In contrast, partial ablation of mushroom body subunits resulting in small Kenyon cell clusters and peduncles was observed very rarely. These findings indicate that hydroxyurea applied during larval stage 1 selectively deletes Kenyon stem cells. The results also show that each mushroom body subunit originates from a very small number of stem cells and develops independently of its neighboring subunit. PMID- 10454375 TI - Metamorphosis of the mushroom bodies; large-scale rearrangements of the neural substrates for associative learning and memory in Drosophila. AB - Paired brain centers known as mushroom bodies are key features of the circuitry for insect associative learning, especially when evoked by olfactory cues. Mushroom bodies have an embryonic origin, and unlike most other brain structures they exhibit developmental continuity, being prominent components of both the larval and the adult CNS. Here, we use cell-type-specific markers, provided by the P[GAL4] enhancer trap system, to follow specific subsets of mushroom body intrinsic and extrinsic neurons from the larval to the adult stage. We find marked structural differences between the larval and adult mushroom bodies, arising as the consequence of large-scale reorganization during metamorphosis. Extensive, though incomplete, degradation of the larval structure is followed by establishment of adult specific alpha and beta lobes. Kenyon cells of embryonic origin, by contrast, were found to project selectively to the adult gamma lobe. We propose that the gamma lobe stores information of relevance to both developmental stages, whereas the alpha and beta lobes have uniquely adult roles. PMID- 10454376 TI - Experience-expectant plasticity in the mushroom bodies of the honeybee. AB - Worker honeybees (Apis mellifera) were reared in social isolation in complete darkness to assess the effects of experience on growth of the neuropil of the mushroom bodies (MBs) during adult life. Comparison of the volume of the MBs of 1 day-old and 7-day-old bees showed that a significant increase in volume in the MB neuropil occurred during the first week of life in bees reared under these highly deprived conditions. All regions of the MB neuropil experienced a significant increase in volume with the exception of the basal ring. Measurement of titers of juvenile hormone JH) in a subset of bees indicated that, as in previous studies, these rearing conditions induced in some bees the endocrine state of high JH associated with foraging, but there was no correlation between JH titer and volume of MB neuropil. Treatment of another subset of dark-reared bees with the JH analog, methoprene, also had no effect of the growth of the MB neuropil. These results demonstrate that there is a phase of MB neuropil growth early in the adult life of bees that occurs independent of light or any form of social interaction. Together with previous findings showing that an increase in MB neuropil volume begins around the time that orientation flights occur and then continues throughout the phase of life devoted to foraging, these results suggest that growth of the MB neuropil in adult bees may have both experience-expectant and experience-dependent components. PMID- 10454377 TI - Spatiotemporal structure of olfactory inputs to the mushroom bodies. AB - A requirement to understand mushroom body (MB) function is to characterize the operations (or transformations) that they impose on incoming signals. Understanding the nature of these integrative operations requires an understanding of the inputs from other brain areas. By inputs we mean not only the anatomical pathways leading to the MBs, but also the dynamic structure of the inflow of sensory (and other) signals. Neurons are complex, capacitative, and generally nonlinear devices that transform barrages of neurochemical packets into electrical waveforms. Their modes of operation are intrinsically time dependent and therefore, their functions or roles in a circuit cannot be inferred only from structural data. Thanks to elegant anatomical, behavioral, genetic, and molecular (for review, see Crittenden et al. 1998; Hammer and Menzel 1998; Heisenberg 1998; Wolf et al. 1998) studies, there is convincing evidence that MB circuits are involved, at least in fruit flies and honeybees, in some forms of odor integration and learning. In vivo electrophysiological studies of MB neurons, however, are rare and mainly restricted to individual (or small populations of) so-called extrinsic neurons, that is, those whose processes link MBs with other brain areas (Schildberger 1983, 1984; Homberg 1984; Hammer 1993; Mauelshagen 1993; Li and Strausfeld 1997). Kaulen et al. (1984) examined extracellular potentials in the MBs of bees, using current source density analysis, and more recently, Laurent and Naraghi (1994) provided a description of stimulus-evoked activity in Kenyon cells (KCs), the intrinsic neurons of the MBs, using intracellular recordings. In this short review we will summarize the recent results from our laboratory in an attempt to provide a description of the spatiotemporal structure of olfactory inputs to the MBs and their intrinsic neurons. We will focus only on the encoding of odor quality. We will then speculate on the possible role of MB circuits for olfactory processing. PMID- 10454378 TI - Integrative properties of the Pe1 neuron, a unique mushroom body output neuron. AB - A mushroom body extrinsic neuron, the Pe1 neuron, connects the peduncle of the mushroom body (MB) with two areas of the protocerebrum in the honeybee brain, the lateral protocerebral lobe (LPL) and the ring neuropil around the alpha-lobe. Each side of the bee brain contains only one Pe1 neuron. Using a combination of intracellular recording and neuroanatomical techniques we analyzed its properties of integrative processing of the different sensory modalities. The Pe1 neuron responds to visual, mechanosensory, and olfactory stimuli. The responses are broadly tuned, consisting of a sustained increase of spike frequency to the onset and offset of light flashes, to horizontal and vertical movements of extended objects, to mechanical stimuli applied to the antennae or mouth parts, and to all olfactory stimuli tested (29 chemicals). These multisensory properties are reflected in its dendritic organization. Serial reconstructions of intracellularly stained Pe1 neurons using confocal microscopy reveal that the Pe1 neuron arborizes throughout all layers of MB peduncle with finger-like, vertically oriented dendrites. The peduncle of the MB is formed by the axons of Kenyon cells, whose dendritic inputs are organized in modality-specific subcompartments of the calyx region. The peduncular arborization indicates that the Pe1 neuron receives input from Kenyon cells of all calycal subcompartments. Because the Pe1 neuron changes its odor responses transiently as a consequence of olfactory learning, we hypothesize that the multimodal response properties might have a role in memory consolidation and help to establish contextual references in the long-term trace. PMID- 10454379 TI - Multiple sites of associative odor learning as revealed by local brain microinjections of octopamine in honeybees. AB - In a classical conditioning procedure, honeybees associate an odor with sucrose resulting in the capacity of the odor to evoke an appetitive response, the extension of the proboscis (PER). Here, we study the effects of pairing an odor with injections of octopamine (OA) as a substitute for sucrose into three putative brain sites of odor/sucrose convergence. OA injected into the mushroom body (MB) calyces or the antennal lobe but not the lateral protocerebral lobe produces a lasting, pairing-specific enhancement of PER. During pairings, OA injected into the MB calyces results in an additional pairing-specific effect, because it does not lead to an acquisition but a consolidation after conditioning. These results suggest that the neuromodulator OA has the capacity of inducing associative learning in an insect brain. Moreover, they suggest the antennal lobes and the calyces as at least partially independent sites of associating odors that may contribute differently to learning and memory consolidation. PMID- 10454380 TI - Dopamine and mushroom bodies in Drosophila: experience-dependent and -independent aspects of sexual behavior. AB - Depletion of dopamine in Drosophila melanogaster adult males, accomplished through systemic introduction of the tyrosine hydroxylase inhibitor 3-iodo tyrosine, severely impaired the ability of these flies to modify their courtship responses to immature males. Mature males, when first exposed to immature males, will perform courtship rituals; the intensity and duration of this behavior rapidly diminishes with time. Dopamine is also required for normal female sexual receptivity; dopamine-depleted females show increased latency to copulation. One kilobase of 5' upstream information from the Drosophila tyrosine hydroxylase (DTH) gene, when fused to the Escherichia coli beta-galactosidase reporter and transduced into the genome of Drosophila melanogaster, is capable of directing expression of the reporter gene in the mushroom bodies, which are believed to mediate learning acquisition and memory retention in flies. Ablation of mushroom bodies by treatment of newly hatched larva with hydroxyurea resulted in the inability of treated mature adult males to cease courtship when placed with untreated immature males. However, functional mushroom bodies were not required for the dopaminergic modulation of an innate behavior, female sexual receptivity. These data suggest that dopamine acts as a signaling molecule within the mushroom bodies to mediate a simple form of learning. PMID- 10454381 TI - Drosophila mushroom bodies are dispensable for visual, tactile, and motor learning. AB - A total of 18 associative learning/memory tests have been applied to Drosophila melanogaster flies lacking mushroom bodies. Only in paradigms involving chemosensory cues as conditioned stimuli have flies been found to be compromised by a block in the mushroom body pathway. Among the learning tasks not requiring these structures are a case of motor learning (yaw torque/heat), a test of the fly's spatial orientation in total darkness, conditioned courtship suppression by mated females, and nine different examples of visual learning. The latter used the reinforcers of heat, visual oscillations, mechanical shaking, or sucrose, and as conditioned stimuli, color, intensity contrast, as well as stationary and moving visual patterns. No forms of consolidated memory have been tested in mushroom body-less flies. With respect to short-term memory the mushroom bodies of Drosophila are specially required for chemosensory learning tasks, but not for associative learning and memory in general. PMID- 10454382 TI - Mushroom bodies suppress locomotor activity in Drosophila melanogaster. AB - Locomotor activity of single, freely walking flies in small tubes is analyzed in the time domain of several hours. To assess the influence of the mushroom bodies on walking activity, three independent noninvasive methods interfering with mushroom body function are applied: chemical ablation of the mushroom body precursor cells; a mutant affecting Kenyon cell differentiation (mushroom body miniature); and the targeted expression of the catalytic subunit of tetanus toxin in subsets of Kenyon cells. All groups of flies with mushroom body defects show an elevated level of total walking activity. This increase is attributable to the slower and less complete attenuation of activity during the experiment. Walking activity in normal and mushroom body-deficient flies is clustered in active phases (bouts) and rest periods (pauses). Neither the initiation nor the internal structure, but solely the termination of bouts seems to be affected by the mushroom body defects. How this finding relates to the well-documented role of the mushroom bodies in olfactory learning and memory remains to be understood. PMID- 10454383 TI - Cancer survival in Britain is poorer than that of her comparable European neighbours. PMID- 10454384 TI - Evidence based palliative care. There is some evidence-and there needs to be more. PMID- 10454385 TI - Doctors and nurses: changing family values? We want to hear about how doctors and nurses work together. PMID- 10454386 TI - What the millennium bug tells us about ourselves. Y2K anxiety is provoked by unlimited ambiguity with a concrete deadline. PMID- 10454387 TI - The joy of being electronic. The BMJ's website is mushrooming. PMID- 10454388 TI - Netherlands published plan for euthanasia law. PMID- 10454389 TI - Immunotherapy produces long term clinical remission. PMID- 10454390 TI - Irish doctors protest over pay and conditions. PMID- 10454391 TI - Life expectancy of Russian men falls to 58. PMID- 10454393 TI - UK hospitals assess eye damage after solar eclipse. PMID- 10454394 TI - Earlier discharge for newborns may increase health risks. PMID- 10454396 TI - Drug treatment beats prison for cutting crime and addiction rates. PMID- 10454395 TI - Global health gap widens says World Bank. PMID- 10454397 TI - Raloxifene can cut fracture risk. PMID- 10454398 TI - Screening of newborn infants for cholestatic hepatobiliary disease with tandem mass spectrometry. AB - OBJECTIVE: To assess the feasibility of screening for cholestatic hepatobiliary disease and extrahepatic biliary atresia by using tandem mass spectrometry to measure conjugated bile acids in dried blood spots obtained from newborn infants at 7-10 days of age for the Guthrie test. SETTING: Three tertiary referral clinics and regional neonatal screening laboratories. DESIGN: Unused blood spots from the Guthrie test were retrieved for infants presenting with cholestatic hepatobiliary disease and from the two cards stored on either side of each card from an index child. Concentrations of conjugated bile acids measured by tandem mass spectrometry in the two groups were compared. MAIN OUTCOME MEASURES: Concentrations of glycodihydroxycholanoates, glycotrihydroxycholanoates, taurodihydroxycholanoates, and taurotrihydroxycholanoates. Receiver operator curves were plotted to determine which parameter (or combination of parameters) would best predict the cases of cholestatic hepatobiliary disease and extrahepatic biliary atresia. The sensitivity and specificity at a selection of cut off values for each bile acid species and for total bile acid concentrations for the detection of the two conditions were calculated. RESULTS: 218 children with cholestatic hepatobiliary disease were eligible for inclusion in the study. Two children without a final diagnosis and five who presented at <14 days of age were excluded. Usable blood spots were obtained from 177 index children and 708 comparison children. Mean concentrations of all four bile acid species were significantly raised in children with cholestatic hepatobiliary disease and extrahepatic biliary atresia compared with the unaffected children (P<0.0001). Of 177 children with cholestatic hepatobiliary disease, 104 (59%) had a total bile acid concentration >33 micromol/l (97.5th centile value for comparison group). Of the 61 with extrahepatic biliary atresia, 47 (77%) had total bile acid concentrations >33 micromol/l. Taurotrihydroxycholanoate and total bile acid concentrations were the best predictors of both conditions. For all cholestatic hepatobiliary disease, a cut off level of total bile acid concentration of 30 micromol/l gave a sensitivity of 62% and a specificity of 96%, while the corresponding values for extrahepatic biliary atresia were 79% and 96%. CONCLUSION: Most children who present with extrahepatic biliary atresia and other forms of cholestatic hepatobiliary disease have significantly raised concentrations of conjugated bile acids as measured by tandem mass spectrometry at the time when samples are taken for the Guthrie test. Unfortunately the separation between the concentrations in these infants and those in the general population is not sufficient to make mass screening for cholestatic hepatobiliary disease a feasible option with this method alone. PMID- 10454400 TI - Perinatal mortality and morbidity among babies delivered in water: surveillance study and postal survey. AB - AIM: To compare perinatal morbidity and mortality for babies delivered in water with rates for babies delivered conventionally (not in water). DESIGN: Surveillance study (of all consultant paediatricians) and postal survey (of all NHS maternity units). SETTING: British Isles (surveillance study); England and Wales (postal survey). SUBJECTS: Babies born in the British Isles between April 1994 and March 1996 who died perinatally or were admitted for special care within 48 hours of birth after delivery in water or after labour in water followed by conventional delivery (surveillance study); babies delivered in water in England and Wales in the same period (postal survey). MAIN OUTCOME MEASURESE Number of deliveries in water in the British Isles that resulted in perinatal death or in admission to special care within 48 hours of birth; and proportions (of such deliveries) of all water births in England and Wales. RESULTS: 4032 deliveries (0.6% of all deliveries) in England and Wales occurred in water. Perinatal mortality was 1.2/1000 (95% confidence interval 0.4 to 2.9) live births; 8.4/1000 (5.8 to 11.8) live births were admitted for special care. No deaths were directly attributable to delivery in water, but 2 admissions were for water aspiration. UK reports of mortality and special care admission rates for babies of women considered to be at low risk of complications during delivery who delivered conventionally ranged from 0.8/1000 (0. 2 to 4.2) to 4.6/1000 (0.1 to 25) live births and from 9.2 (1.1 to 33) to 64/1000 (58 to 70) live births respectively. Compared with regional data for low risk, spontaneous, normal vaginal deliveries at term, the relative risk for perinatal mortality associated with delivery in water was 0.9 (99% confidence interval 0.2 to 3.6). CONCLUSIONS: Perinatal mortality is not substantially higher among babies delivered in water than among those born to low risk women who delivered conventionally. The data are compatible with a small increase or decrease in perinatal mortality for babies delivered in water. PMID- 10454399 TI - Population based study of social and productive activities as predictors of survival among elderly Americans. AB - OBJECTIVES: To examine any association between social, productive, and physical activity and 13 year survival in older people. DESIGN: Prospective cohort study with annual mortality follow up. Activity and other measures were assessed by structured interviews at baseline in the participants' homes. Proportional hazards models were used to model survival from time of initial interview. SETTING: City of New Haven, Connecticut, United States. PARTICIPANTS: 2761 men and women from a random population sample of 2812 people aged 65 and older. MAIN OUTCOME MEASURE: Mortality from all causes during 13 years of follow up. RESULTS: All three types of activity were independently associated with survival after age, sex, race/ethnicity, marital status, income, body mass index, smoking, functional disability, and history of cancer, diabetes, stroke, and myocardial infarction were controlled for. CONCLUSIONS: Social and productive activities that involve little or no enhancement of fitness lower the risk of all cause mortality as much as fitness activities do. This suggests that in addition to increased cardiopulmonary fitness, activity may confer survival benefits through psychosocial pathways. Social and productive activities that require less physical exertion may complement exercise programmes and may constitute alternative interventions for frail elderly people. PMID- 10454401 TI - Polymorphism in high density lipoprotein paraoxonase gene and risk of acute myocardial infarction in men: prospective nested case-control study. PMID- 10454402 TI - The screaming man. PMID- 10454404 TI - Results are unlikely to be as good in routine practice. PMID- 10454403 TI - Open randomised trial of intermittent very low energy diet together with nicotine gum for stopping smoking in women who gained weight in previous attempts to quit. AB - OBJECTIVE: To determine whether attempts to prevent weight gain will increase success rates for stopping smoking. DESIGN: 16 week, open, randomised study with 1 year follow up. SETTING: Obesity unit. SUBJECTS: 287 female smokers who had quit smoking before but started again because of weight concerns. INTERVENTION: Combination of a standard smoking cessation programme with nicotine gum and a behavioural weight control programme including a very low energy diet. A control group was treated with the identical programme but without the diet. MAIN OUTCOME MEASURE: Sustained cessation of smoking. RESULTS: After 16 weeks, 68/137 (50%) women had stopped smoking in the diet group versus 53/150 (35%) in the control group (P=0.01). Among these women, weight fell by mean 2.1 (95% confidence interval 2.9 to 1.3) kg in the diet group but increased by 1.6 (0.9 to 2.3) kg in the control group (P<0.001). After 1 year the success rates in the diet and control groups were 38/137 (28%) and 24/150 (16%) respectively (P<0.05), but there was no statistical difference in weight gain. CONCLUSIONS: Combining the smoking cessation programme with an intervention to control weight helped women to stop smoking and control weight. PMID- 10454406 TI - The qat party. PMID- 10454407 TI - ABC of intensive care. Cutting edge. PMID- 10454405 TI - Stable chronic obstructive pulmonary disease. PMID- 10454408 TI - Cancer in the developing world: a call to action. PMID- 10454409 TI - Methods in health service research. An introduction to bayesian methods in health technology assessment. PMID- 10454410 TI - Contacting GP surgeries over Christmas. I will be in my surgery on Christmas day if ministers and researchers are at work too. PMID- 10454411 TI - Hyponatraemia after orthopaedic surgery. Failsafe system is needed. PMID- 10454412 TI - Intensive care is not expensive compared with other treatments. PMID- 10454413 TI - Orbital trauma. Antibiotic prophylaxis needs to be given only in certain circumstances. PMID- 10454414 TI - Twins and the fetal origins hypothesis. Many variables differ between twins and singleton infants. PMID- 10454416 TI - "Black boxing:" response by editor of hospital doctor. PMID- 10454415 TI - Hand washing. Why I don't wash my hands between each patient contact. PMID- 10454417 TI - Referral to intensive care units should not be limited to consultants. PMID- 10454418 TI - Shakespeare knew the layered clothing sign of schizophrenia. PMID- 10454420 TI - Charles Peter Broad. PMID- 10454419 TI - Evaluation of NHS direct. Too early to draw conclusions. PMID- 10454421 TI - QBase: Anaesthesia: 1. PMID- 10454423 TI - Water births. PMID- 10454422 TI - "Unselling" the cigarette. PMID- 10454425 TI - Fancy drugs for worried folks. PMID- 10454424 TI - A patient's eye view of quality. PMID- 10454427 TI - Single screening test for neonatal cholestasis is not yet feasible from blood spots. PMID- 10454426 TI - A global look at cancer. PMID- 10454429 TI - Babies delivered in water: perinatal mortality is no higher. PMID- 10454428 TI - Social activities lengthen life in elderly Americans. PMID- 10454430 TI - A single genetic polymorphism may account for a fifth of infarctions. PMID- 10454431 TI - Weight control measures help women stop smoking. PMID- 10454432 TI - What should the developing world do about cancer? PMID- 10454433 TI - Council honorees and the Nobel Prize. Our continued anniversary celebration. PMID- 10454434 TI - Harriet Pearson Dustan. PMID- 10454435 TI - Kallikrein gene delivery inhibits vascular smooth muscle cell growth and neointima formation in the rat artery after balloon angioplasty. AB - Tissue kallikrein cleaves kininogen substrate to produce vasoactive kinin peptides that have been implicated in the proliferation of vascular smooth muscle cells (VSMCs). To explore potential roles of the kallikrein-kinin system in vascular biology, we evaluated the effects of adenovirus-mediated human kallikrein gene delivery on the growth of primary cultured VSMCs and in balloon injured rat artery in vivo. Kallikrein gene transfer into cultured rat VSMCs resulted in time-dependent secretion of recombinant human tissue kallikrein and inhibition of cell proliferation. Balloon angioplasty reduced endogenous rat tissue kallikrein mRNA and protein levels at the injured site. In rats that received adenovirus-mediated human kallikrein gene delivery, we observed a 39% reduction in intima/media ratio at the injured vessel after delivery compared with that of rats that received control virus (n=8, P<0.01). Icatibant, a specific bradykinin B(2) receptor antagonist, blocked the protective effect and reversed the intima/media ratio to that of the control rats (n=5, P<0.01). After gene delivery, human kallikrein mRNA was identified at the injured vessel and a 3 fold increase occurred in kininogenase activity. cAMP and cGMP levels in balloon injured aorta increased significantly at 4, 7, and 14 days after kallikrein gene delivery, but icatibant abolished the increase. These results provide new insights into the role of the vascular kallikrein-kinin system and have significant implications for gene therapy to treat restenosis or atherosclerosis. PMID- 10454436 TI - Angiotensin-converting enzyme inhibition modifies angiotensin but not kinin peptide levels in human atrial tissue. AB - Angiotensin-converting enzyme (ACE) converts angiotensin I (Ang I) to angiotensin II (Ang II) and metabolizes bradykinin and kallidin peptides. Decreased Ang II levels and increased kinin peptide levels are implicated in the mediation of the therapeutic effects of ACE inhibition. However, alternative non-ACE pathways of Ang II formation have been proposed to predominate in human heart. We investigated the effects of ACE inhibition on cardiac tissue levels of angiotensin and kinin peptides. High-performance liquid chromatography-based radioimmunoassays were used to measure angiotensin peptides and hydroxylated and nonhydroxylated bradykinin and kallidin peptides in right atrial appendages of subjects who had been prepared for cardiopulmonary bypass. Peptide levels in subjects who received ACE inhibitor therapy were compared with those who did not receive ACE inhibitor therapy. ACE inhibition reduced Ang II levels, which was associated with an 80% reduction in the Ang II/Ang I ratio. ACE inhibition did not modify either bradykinin or kallidin peptide levels or the bradykinin-(1 7)/bradykinin-(1-9) ratio. The 80% reduction in the Ang II/Ang I ratio by ACE inhibition indicated a primary role for ACE in the conversion of Ang I to Ang II in atrial tissue. These data support a role for reduced Ang II levels but do not support a role for increased kinin peptide levels in mediating the direct cardiac effects of ACE inhibition. PMID- 10454437 TI - Early onset salt-sensitive hypertension in bradykinin B(2) receptor null mice. AB - Kinins have been implicated in the hemodynamic adaptation to postnatal life. The present study examined the impact of bradykinin B(2) receptor (B(2)R) gene disruption on the postnatal changes in blood pressure (BP) and the susceptibility to early onset salt-sensitive hypertension in mice. B(2)R null (-/-) and wild type (+/+) mice were fed normal (NS, 1% NaCl) or high (HS, 5% NaCl) salt diets during pregnancy. After birth, the pups remained with their mothers until they were weaned and were subsequently continued on the respective maternal salt intake until 4 months of age. The age-related changes at 3 and 4 months in tail cuff BP and anesthetized mean arterial pressure at 4 months were not different in NS/B(2)R(-/-) and NS/B(2)R(+/+) mice. However, there was a mild increase in BP in NS/B(2)R(-/-) at 2 months versus NS/B(2)R(+/+). In contrast, HS/B(2)R(-/-) mice manifested early onset and persistent elevations of tail-cuff BP (P<0.05) at 2, 3, and 4 months versus other groups. MAP was also higher in HS/B(2)R(-/-) than HS/B(2)R(+/+), NS/B(2)R(-/-), and NS/B(2)R(+/+) (91+/-3 versus 75+/-5, 74+/-2, and 70+/-2 mm Hg, respectively; P<0.05). Kidney renin and angiotensin type 1 receptor mRNA levels were not different. Additional studies showed that a delay in the initiation of HS until after birth was accompanied by later development of hypertension, although postnatal discontinuation of HS resulted in a gradual return of BP to normal values by 4 months of age. The results demonstrate that (1) kinins protect the developing animal from salt-sensitive hypertension, (2) lack of B(2)R from early development does not alter the maturation of BP under conditions of normal sodium intake, and (3) exposure to a HS diet during fetal life is not sufficient in itself to induce long-term hypertension in either wild type or B(2)R null mice. PMID- 10454438 TI - Effects of potassium on blood pressure in salt-sensitive and salt-resistant black adolescents. AB - This study examined the effects of increasing dietary potassium on ambulatory blood pressure nondipping status (<10% decrease in blood pressure from awake to asleep) and cardiovascular reactivity in salt-sensitive and salt-resistant black adolescents. A sample of 58 normotensive (blood pressure, 101/57+/-9/4 mm Hg) black adolescents (aged 13 to 16 years) participated in a 5-day low sodium diet (50 mmol/24 h) followed by a 10-day high sodium diet (150 mmol/24 h NaCl supplement) to determine salt-sensitivity status. Participants showed a significant increase in urinary sodium excretion (24+/-19 to 224+/-65 mmol/24 h) and were identified as salt-sensitive if their mean blood pressure increase was >/=5 mm Hg from the low to high sodium diet. Sixteen salt-sensitive and 42 salt resistant subjects were then randomly assigned to either a 3-week high potassium diet (80 mmol/24 h) or usual diet control group. Urinary potassium excretion significantly increased in the treatment group (35+/-7 to 57+/-21 mmol/24 h). At baseline, a significantly greater percentage of salt-sensitive (44%) compared with salt-resistant (7%) subjects were nondippers on the basis of diastolic blood pressure classifications (P<0.04). After the dietary intervention, all of the salt-sensitive subjects in the high potassium group achieved dipper status as a result of a drop in nocturnal diastolic blood pressure (daytime, 69 versus 67 mm Hg; nighttime, 69 versus 57 mm Hg). No significant group differences in cardiovascular reactivity were observed. These results suggest that a positive relationship between dietary potassium intake and blood pressure modulation can still exist even when daytime blood pressure is unchanged by a high potassium diet. PMID- 10454439 TI - Genetic isolation of a chromosome 1 region affecting susceptibility to hypertension-induced renal damage in the spontaneously hypertensive rat. AB - Linkage studies in the fawn-hooded hypertensive rat have suggested that genes influencing susceptibility to hypertension-associated renal failure may exist on rat chromosome 1q. To investigate this possibility in a widely used model of hypertension, the spontaneously hypertensive rat (SHR), we compared susceptibility to hypertension-induced renal damage between an SHR progenitor strain and an SHR congenic strain that is genetically identical except for a defined region of chromosome 1q. Backcross breeding with selection for the markers D1Mit3 and Igf2 on chromosome 1 was used to create the congenic strain (designated SHR.BN-D1Mit3/Igf2) that carries a 22 cM segment of chromosome 1 transferred from the normotensive Brown Norway rat onto the SHR background. Systolic blood pressure (by radiotelemetry) and urine protein excretion were measured in the SHR progenitor and congenic strains before and after the induction of accelerated hypertension by administration of DOCA-salt. At the same level of DOCA-salt hypertension, the SHR.BN-D1Mit3/Igf2 congenic strain showed significantly greater proteinuria and histologically assessed renal vascular and glomerular injury than the SHR progenitor strain. These findings demonstrate that a gene or genes that influence susceptibility to hypertension-induced renal damage have been trapped in the differential chromosome segment of the SHR.BN D1Mit3/Igf2 congenic strain. This congenic strain represents an important new model for the fine mapping of gene(s) on chromosome 1 that affect susceptibility to hypertension-induced renal injury in the rat. PMID- 10454440 TI - Inhibition of the p53 tumor suppressor gene results in growth of human aortic vascular smooth muscle cells. Potential role of p53 in regulation of vascular smooth muscle cell growth. AB - Loss of activity of the p53 tumor suppressor gene product has been postulated in the pathogenesis of human restenosis. Although the antioncogenes p53 and retinoblastoma (Rb) susceptibility gene have been reported to play a pivotal role in cell cycle progression in various cells, the role of p53 and Rb in the growth of human vascular smooth muscle cells (VSMC) has not yet been clarified. We used antisense strategy against p53 and Rb genes by the viral envelope-liposomal method. Transfection of antisense p53 oligodeoxynucleotides (ODN) alone resulted in an increase in DNA synthesis compared with control (P<0.01). Similarly, transfection of antisense Rb ODN alone resulted in a higher DNA synthesis rate than control (P<0.01). Moreover, increase in VSMC number was only induced by transfection of antisense p53 ODN alone or cotransfection of p53/Rb ODN (P<0.01), whereas a single transfection of antisense Rb ODN had little effect on cell number. Therefore, we hypothesized that this discrepancy is due to the induction of apoptosis mediated by p53. Interestingly, apoptotic cells were markedly increased in VSMC transfected with antisense Rb ODN alone, accompanied by the induction of p53 protein. The number of apoptotic cells was attenuated by cotransfection of antisense p53 ODN (P<0.01). We finally examined the molecular mechanisms of apoptosis induced by the absence of Rb. In VSMC transfected with antisense Rb ODN, bax, a promoter of apoptosis, was significantly increased in VSMC transfected with antisense Rb ODN (P<0.01), whereas bcl-2 and Fas did not play a pivotal role in the induction of apoptosis. Overall, these data first demonstrated that the antioncogenes p53 and Rb negatively regulated the cell cycle in VSMC, suggesting that the modulation of their activity may mediate VSMC growth such as that in restenosis and atherosclerosis. The presence of p53 plays a pivotal role in the regulation of apoptosis in human VSMC growth, probably through the bax pathway. These results provide evidence that p53 is a functional link between cell growth and apoptosis in VSMC. PMID- 10454441 TI - Arterial stiffness and the development of hypertension. The ARIC study. AB - Decreased elasticity in large and medium-sized arteries has been postulated to be associated with cardiovascular diseases. We prospectively examined the relation between arterial elasticity and the development of hypertension over 6 years of follow-up in a cohort of 6992 normotensive men and women aged 45 to 64 years at baseline from the biracial, population-based Atherosclerosis Risk in Communities (ARIC) Study. Arterial elasticity was measured from high-resolution B-mode ultrasound examination of the left common carotid artery as adjusted arterial diameter change (in micrometers, simultaneously adjusted for diastolic blood pressure, pulse pressure, pulse pressure squared, diastolic arterial diameter, and height), Peterson's elastic modulus (in kilopascals), Young's elastic modulus (in kilopascals), and beta stiffness index. Incident hypertension (n=551) was defined as systolic blood pressure >/=160 mm Hg, diastolic blood pressure >/=95 mm Hg, or the use of antihypertensive medication at a follow-up examination conducted every 3 years. The age-, ethnicity-, center-, gender-, education-, smoking-, heart rate-, and obesity-adjusted means (SE) of baseline adjusted arterial diameter change, Peterson's elastic modulus, Young's elastic modulus, and beta stiffness index were 397 (5), 148 (2.0), 787 (12.7), and 11.43 (0.16), respectively, in persons who developed hypertension during follow-up, in contrast to 407 (1), 124 (0.6), 681 (3.7), and 10.34 (0.05), respectively, for persons who did not. The similarly adjusted cumulative incident rates of hypertension from the highest to the lowest quartiles of arterial elasticity were 6.7%, 8.0%, 7.3%, and 9.6%, respectively, when measured by adjusted arterial diameter change (P<0.01). One standard deviation decrease in arterial elasticity was associated with 15% greater risk of hypertension, independent of established risk factors for hypertension and the level of baseline blood pressure. These results suggest that lower arterial elasticity is related to the development of hypertension. PMID- 10454442 TI - Effects of aging and antihypertensive treatment on aortic internal diameter in spontaneously hypertensive rats. AB - The effect of antihypertensive treatment on the development of large-artery remodeling in young animals has been widely studied, but reversal of established changes in older hypertensive animals has been largely ignored, although the latter represents a better paradigm for the human condition. We studied the effect of treatment with captopril plus hydrochlorothiazide, from 3 months onward, on geometry and wall stress of the thoracic aorta of adult (9 months, maturation) and old (15 months, senescence) spontaneously hypertensive rats; normotensive Wistar-Kyoto rats were used as controls. At 3 months of age, blood pressure, medial cross-sectional area, and internal diameter were higher in spontaneously hypertensive rats than in Wistar-Kyoto rats. In both strains, medial cross-sectional area and lumen diameter increased during maturation; there was little change with senescence. Changes in blood pressure were minor. Because medial hypertrophy failed to compensate for the wider lumen and higher intraluminal pressure in spontaneously hypertensive rats, medial stress was higher in these rats than in Wistar-Kyoto rats. Captopril plus hydrochlorothiazide rapidly lowered blood pressure and medial cross-sectional area. Despite a marked fall in blood pressure, the internal diameter of the thoracic aorta of treated animals was similar to that of untreated animals after 6 months of treatment and started to fall only after the animals had been treated for 1 year. Thus, under treatment with captopril plus hydrochlorothiazide, medial stress remained elevated, even after very-long-term treatment, because medial cross-sectional area was not adapted to internal diameter. We suggest that some changes in large-artery structure associated with hypertension and aging, such as the increase in diameter, take considerable time to regress after blood pressure is lowered, and this may explain why, despite treatment, wall stress remains elevated. PMID- 10454443 TI - Microvascular responses to ischemia/reperfusion in normotensive and hypertensive rats. AB - The objective of the present study was to determine whether long-term arterial hypertension renders the microvasculature more vulnerable to the deleterious inflammatory responses elicited by ischemia and reperfusion (I/R). Intravital fluorescence microscopy was used to monitor leukocyte adherence and emigration, platelet-leukocyte aggregation, and albumin extravasation in mesenteric postcapillary venules of spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY) after 10 minutes of ischemia and subsequent reperfusion. Significant and comparable increases in leukocyte adherence/emigration and the formation of platelet aggregates were elicited by I/R in both WKY and SHR. Albumin extravasation was enhanced after I/R in SHR, but not in WKY. Monoclonal antibodies directed against the adhesion glycoproteins CD18, P-selectin, or ICAM 1 showed similar patterns of protection against the I/R-induced inflammatory responses in WKY and SHR. The enhanced albumin extravasation noted in postischemic venules of SHR was prevented by immunoneutralization of either CD18 on leukocytes or ICAM-1 on endothelial cells. These results suggest that, whereas long-term arterial hypertension does not significantly modify the leukocyte and platelet recruitment normally elicited in venules by I/R, it does result in an exaggerated albumin leakage response, which is mediated by an interaction between beta(2) (CD18) integrins on leukocytes and ICAM-1 on endothelial cells. PMID- 10454444 TI - Evaluation of common carotid hemodynamic forces. Relations with wall thickening. AB - The localization of atherosclerotic lesions is influenced by hemodynamic factors, namely, shear stress and tensive forces. The present study investigated the relationships between shear stress and circumferential wall tension and between these hemodynamic factors and the intima-media thickness (IMT) of the common carotid artery in healthy men. Fifty-eight subjects were studied. Shear stress was calculated as blood viscosityxblood velocity/internal diameter. Circumferential wall tension was calculated as blood pressurexinternal radius. Blood velocity, internal diameter, and IMT were measured by high-resolution echo Doppler. Mean shear stress was 12.6+/-3.3 dynes/cm(2) (mean+/-SD; range, 4.8 to 20.4) and was inversely related with age, blood pressure, and body mass index (BMI). Mean circumferential wall tension was 3.4+/-0.6x10(4) dynes/cm (range 2.4 to 5.6) and was directly associated with age and BMI. IMT was inversely associated with shear stress (r=0.55, P<0. 0001) and directly associated with circumferential wall tension (r=0. 43, P<0.0001). Shear stress and circumferential wall tension were inversely correlated (r=0.66, P<0.0001). In multiple regression analysis, shear stress and (marginally) cholesterol were independently associated with IMT, whereas circumferential wall tension, age, and BMI were not. These findings confirm that common carotid shear stress varies among healthy individuals and decreases as age, blood pressure, and BMI increase. Our findings also demonstrate that circumferential wall tension is directly associated with wall thickness, age, and BMI and that shear stress is associated with common carotid IMT independent of other hemodynamic, clinical, or biochemical factors. PMID- 10454445 TI - Impaired isoproterenol-induced hyperpolarization in isolated mesenteric arteries of aged rats. AB - Stimulation of vascular beta-adrenoceptors leads to membrane hyperpolarization, presumably via the beta-adrenoceptor/G(s) protein/adenylate cyclase signaling cascade; the ionic mechanisms of this phenomenon remain unclear. beta Adrenoceptor-mediated vascular relaxation is impaired with aging; however, little is known concerning whether beta-adrenoceptor-mediated hyperpolarization is altered with aging. We sought to determine the ionic mechanisms of isoproterenol induced hyperpolarization in the rat mesenteric resistance artery, as well as the age-related changes in isoproterenol-induced hyperpolarization and their underlying mechanisms. Isoproterenol-induced hyperpolarization was inhibited by high-K(+) solution and glibenclamide (10(-6) mol/L), an inhibitor of ATP sensitive K(+) channels (K(ATP)), but not by apamin, iberiotoxin, or charybdotoxin, inhibitors of Ca(2+)-activated K(+) channels. Isoproterenol induced hyperpolarization was markedly less in aged rats (>/=24 months) than in adults rats (12 to 20 weeks) (3x10(-6) mol/L; -3.1 versus -9.9 mV; P<0.001; n=8 to 9). Cholera toxin (10(-9) g/mL), an activator of G(s), evoked hyperpolarization only in adult rats. Hyperpolarization to forskolin, a direct activator of adenylate cyclase, was also reduced to some extent in aged rats (10( 5) mol/L; -8.8 versus -13 mV; P<0.05; n=6), whereas hyperpolarization to levcromakalim, a K(ATP) opener, was comparable in both groups. These findings suggest that isoproterenol elicits hyperpolarization via an opening of K(ATP) in the rat resistance artery and that isoproterenol-induced hyperpolarization is attenuated in aged rats mainly because of a defective coupling of beta adrenoceptors to adenylate cyclase and partly because of a defect at the level of adenylate cyclase, but not because of an alteration of K(ATP) per se. PMID- 10454446 TI - Apoptosis during regression of cardiac hypertrophy in spontaneously hypertensive rats. Temporal regulation and spatial heterogeneity. AB - We previously reported that increased apoptosis participates in the regression of aortic hypertrophy in spontaneously hypertensive rats. To further document the potential role of apoptosis in cardiovascular therapy, we examined apoptosis during regression of hypertrophy in the heart of spontaneously hypertensive rats receiving the antihypertensive drug enalapril (30 mg. kg(-1). d(-1)), losartan (30 mg. kg(-1). d(-1)), nifedipine (35 mg. kg(-1). d(-1)), hydralazine (40 mg. kg(-1). d(-1)), propranolol (50 mg. kg(-1). d(-1)), or hydrochlorothiazide (75 mg. kg(-1). d(-1)) for 1 to 4 weeks, starting at 10 to 11 weeks of age. Systolic blood pressure and heart rate were measured by the tail-cuff method. Markers of apoptosis included oligonucleosomal DNA fragmentation in extracted cardiac DNA or in situ in ventricular cross sections labeled with terminal deoxynucleotidyl transferase. Cardiac DNA synthesis was evaluated by [(3)H]-thymidine incorporation in vivo. All drugs reduced cardiac workload, defined as the product of blood pressure and heart rate, by >20% at 4 weeks. However, only nifedipine, enalapril, losartan, and propranolol reduced cardiac mass (>19%) within 4 weeks. Regression of cardiac hypertrophy was accompanied by a 50% to 300% increase in DNA fragmentation and a >20% reduction in DNA synthesis, resulting in a >20% reduction in cardiac DNA content after 4 weeks. Apoptosis induction occurred early and was transient within 4 weeks of nifedipine, enalapril, or losartan administration. With all regression-inducing drugs, the increase in DNA fragmentation occurred mainly in the subepicardium. Thus, transient induction of apoptosis in the subepicardium appears to be a characteristic feature of the early response to drug-induced regression of cardiac hypertrophy in spontaneously hypertensive rats. PMID- 10454447 TI - Validity of echocardiographic measurement in an epidemiological study. Project HeartBeat! AB - In Project HeartBeat!, a longitudinal study of cardiovascular disease risk factors in healthy children and adolescents, 3 samples of 40, 80, and 182 echocardiograms, respectively, were randomly selected and reread to evaluate intraobserver and interobserver variabilities and comparability between measurements of field echocardiographic technicians and reference readings at Texas Children's Hospital. Included in the evaluation were 8 M-mode echocardiographic measurements, ie, aortic root diameter, left atrial diameter, and end-diastolic and end-systolic measurements of interventricular septal thickness, left ventricular (LV) diameter, and LV posterior wall thickness; 8 Doppler measurements; and a calculated LV mass. Means and SDs of the differences of the paired measurements were used to assess the relative bias and random error of the measurements. For the intraobserver comparison, means and SDs of the differences were very small, indicating that the echo measurements were performed consistently by each project echo technician. Interobserver comparison showed statistically but not clinically significant differences between the paired readings of end-diastolic septal thickness, end-systolic LV posterior wall thickness, and 5 Doppler measurements. Comparison with reference readings at Texas Children's Hospital showed significant differences in diastolic LV diameter, systolic septal thickness, and right ventricular ejection time. These differences, however, were minimal with limited clinical significance. Mean differences in LV mass for the corresponding comparisons were -1.82, 4.50, and 0.0013 g, and the SDs were 18.79, 24.16, and 12.35 g, respectively. We conclude that the echocardiographic measurements taken from healthy children in a longitudinal study can be made accurately with acceptable reproducibility. PMID- 10454448 TI - Autonomic modulation and QT interval dispersion in hypertensive subjects with anxiety. AB - Anxiety is associated with an increased risk of sudden death. QT dispersion is a marker of cardiac repolarization instability and is seen in conditions of high risk of sudden death. The purpose of this study was to evaluate autonomic nervous system control and QT dispersion in hypertensive subjects with anxiety symptoms. In a recent preliminary study, we observed that hypertensive individuals reporting high scores on a self-assessment anxiety scale had more marked left ventricular hypertrophy. In 105 hypertensive subjects divided into 3 groups according to severity of anxiety, we evaluated autonomic control by short-term power spectral analysis of RR and arterial pressure variability at rest (baseline) and during sympathetic stress (tilt test), left ventricular mass index, and heart rate-corrected QT (QTc) dispersion. At baseline, hypertensive subjects with higher anxiety symptom scores had significantly lower high frequency RR values expressed in absolute terms (P<0.05) and in normalized units (P<0.05) than their counterparts without anxiety symptoms. Hypertensive subjects with anxiety also had a higher mean left ventricular mass index (P<0.001) and greater QTc dispersion (P<0.001). Both indexes and high frequency (P<0.05) correlated with severity of anxiety. These findings suggest that anxiety is associated with autonomic imbalance. This condition could favor an increase in left ventricular mass. Myocardial hypertrophy alone or combined with neuroautonomic imbalance may lead to QT dispersion. PMID- 10454449 TI - Effect of magnesium deficiency on autonomic circulatory regulation in conscious rats. AB - A close relationship between magnesium and cardiovascular function has been reported; however, the effect of magnesium deficiency on autonomic cardiovascular regulation has not been clarified. We investigated the effect of magnesium deficiency on the autonomic regulation of oscillations of the R-R interval, arterial blood pressure (BP), and renal sympathetic nerve activity (RSNA) by using the maximum entropy method in conscious rats. Its effect on baroreflex control of RSNA and heart rate were also investigated with a logistic function curve. Mean BP in magnesium-deficient rats was higher than that in control rats (mean+/-SE, 114.0+/-4.3 versus 101.6+/-3.4 mm Hg; P<0.05), and urinary excretion of catecholamine was increased by 2.4-fold. The fraction of low-frequency oscillation of RSNA was reduced (31.7+/-0.9% versus 36.2+/-1.5%, P<0.05) and the correlation between low-frequency oscillations of BP and RSNA was weakened in magnesium-deficient rats. There was no difference in high-frequency oscillation of the R-R interval, which is related to vagal tone, whereas sympathetic tone became dominant (square root of low-frequency/high-frequency ratio of R-R interval, 1.00+/-0.05 versus 0.67+/-0.05, P<0.0001) in magnesium-deficient rats. The maximal gain in the BP-RSNA relation tended to be reduced in magnesium deficient rats (-7.7+/-1.1% versus -12.2+/-1.9%/mm Hg, P=0. 07); however, that in the BP-heart rate relation was increased (-8. 1+/-0.7 versus -4.5+/-0.5 bpm/mm Hg, P<0.01). These results suggest that magnesium deficiency induces sympathetic excitation, which results in hypertension but attenuates the baroreflex-related response of sympathetic nerves, whereas magnesium deficiency enhances the sensitivity of the sinus node to autonomic regulation. PMID- 10454450 TI - Docosahexaenoic acid but not eicosapentaenoic acid lowers ambulatory blood pressure and heart rate in humans. AB - Animal studies suggest that the 2 major omega3 fatty acids found in fish, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), may have differential effects on blood pressure (BP) and heart rate (HR). The aim of this study was to determine whether there were significant differences in the effects of purified EPA or DHA on ambulatory BP and HR in humans. In a double-blind, placebo controlled trial of parallel design, 59 overweight, mildly hyperlipidemic men were randomized to 4 g/d of purified EPA, DHA, or olive oil (placebo) capsules and continued their usual diets for 6 weeks. Fifty-six subjects completed the study. Only DHA reduced 24-hour and daytime (awake) ambulatory BP (P<0.05). Relative to the placebo group, 24-hour BP fell 5.8/3.3 (systolic/diastolic) mm Hg and daytime BP fell 3.5/2.0 mm Hg with DHA. DHA also significantly reduced 24 hour, daytime, and nighttime (asleep) ambulatory HRs (P=0. 001). Relative to the placebo group, DHA reduced 24-hour HR by 3. 5+/-0.8 bpm, daytime HR by 3.7+/-1.2 bpm, and nighttime HR by 2. 8+/-1.2. EPA had no significant effect on ambulatory BP or HR. Supplementation with EPA increased plasma phospholipid EPA from 1. 66+/ 0.07% to 9.83+/-0.06% (P<0.0001) but did not change DHA levels. Purified DHA capsules increased plasma phospholipid DHA levels from 4.00+/-0.27% to 10.93+/ 0.62% (P<0.0001) and led to a small, nonsignificant increase in EPA (1.52+/-0.12% to 2.26+/-0.16%). Purified DHA but not EPA reduced ambulatory BP and HR in mildly hyperlipidemic men. The results of this study suggest that DHA is the principal omega3 fatty acid in fish and fish oils that is responsible for their BP- and HR lowering effects in humans. These results have important implications for human nutrition and the food industry. PMID- 10454451 TI - Multiple clinic and home blood pressure measurements versus ambulatory blood pressure monitoring. AB - To compare multiple clinic and home blood pressure (BP) measurements and ambulatory BP monitoring in the clinical evaluation of hypertension, we studied 239 middle-aged pharmacologically untreated hypertensive men and women who were referred to the study from the primary healthcare provider. Ambulatory BP monitoring was successfully completed for 233 patients. Clinic BP was measured by a trained nurse with a mercury sphygmomanometer and averaged over 4 duplicate measures. Self-recorded home BP was measured with a semiautomatic oscillometric device twice every morning and twice every evening on 7 consecutive days. Ambulatory BP was recorded with an auscultatory device. Two-dimensionally controlled M-mode echocardiography was successfully performed on 232 patients. Twenty-four-hour urinary albumin was determined by nephelometry. Clinic BP was 144.5+/-12.6/94.5+/-7.4 mm Hg, home BP (the mean of 14 self-recorded measures) was 138.9+/-13.1/92.9+/-8.6 mm Hg, home morning BP (the mean of the first 4 duplicate morning measures) was 137.1+/-13.7/92.4+/-9.2 mm Hg, daytime ambulatory BP was 148.3+/-13. 9/91.9+/-7.8 mm Hg, nighttime ambulatory BP was 125.5+/ 16.4/75. 6+/-8.9 mm Hg, and 24-hour ambulatory BP was 141.7+/-14.0/87.2+/-7.6 mm Hg. Pearson correlation coefficients of clinic, home, home morning, and daytime ambulatory BPs to albuminuria and to the characteristics of the left ventricle were nearly equal. In multivariate regression analyses, 36% (P<0.0001) of the cross-sectional variation in left ventricular mass index was attributed to gender and home morning systolic BP in models that originally included age, gender, and clinic, self-measured home morning, and ambulatory daytime, nighttime, and 24 hour systolic and diastolic BPs. We concluded that carefully controlled nonphysician-measured clinic and self-measured home BPs, when averaged over 4 duplicate measurements, are as reliable as ambulatory BP monitoring in the clinical evaluation of untreated hypertension. PMID- 10454452 TI - Diagnosis of white coat hypertension by ambulatory blood pressure monitoring. AB - White coat hypertension (WCH) is common in referred hypertensive patients. Ambulatory blood pressure monitoring (ABPM) is not free from the white coat syndrome. We examined the use of the elevation of the first and last measurements of ABPM for diagnosis of WCH in a hypertensive population that had been referred to a hospital-based hypertension unit. Data were obtained on 1350 patients for clinic and ABPM parameters. WCH, as diagnosed by conventional clinic blood pressure (BP) measurement, was compared with a variety of alternative methods determined from ABPM. In all cases, mean daytime pressure was <135 mm Hg/85 mm Hg with an elevation of clinic BP >/=140 mm Hg systolic or 90 mm Hg diastolic. The definitions tested for this elevation were first hour mean pressure, first reading, maximum reading in first hour, last hour mean pressure, last reading, maximum reading in the last hour and maximum reading in first or last hour. Elevation of the maximum pressure in the first hour or last hour above 140 mm Hg systolic or 90 mm Hg diastolic showed a high level of agreement (kappa=0.91) with classical WCH for diagnosis of the white coat syndrome. Termed ambulatory white coat hypertension, patients with this finding were older than classic white coat patients and had higher daytime (127+/-6/78+/-5 mm Hg versus 121+/-5.5/74+/-6 mm Hg, P<0.005 for systolic and diastolic) and nighttime (114+/-11/67+/-8 mm Hg versus 106+/-9/61+/-6 mm Hg, P<0.005 for systolic and diastolic) pressures. They also had a significantly greater Sokolow-Lyon index (leads V(1)+V(5), 21+/-7 mV versus 18+/-6 mV). Elevation of BP above 140 mm Hg systolic or 90 mm Hg diastolic in the first or last hour of monitoring diagnoses patients with a white coat response in whom there is a higher BP profile than in patients with classic white coat response alone. We suggest, therefore, that this is a better measure of the white coat phenomenon. PMID- 10454453 TI - Differential effects of T- and L-type calcium antagonists on glomerular dynamics in spontaneously hypertensive rats. AB - To determine whether there is a difference in the effects of T- and L-type calcium antagonists on systemic, renal, and glomerular hemodynamics, the pathological changes of N(G)-nitro-L-arginine methyl ester (L-NAME)-exacerbated nephrosclerosis and clinical alterations were investigated in spontaneously hypertensive rats (SHR). Seven groups of 17-week-old male SHRs were studied: Group 1, control; Group 2, mibefradil, 50 mg. kg(-1). d(-1); Group 3, L-NAME in drinking water, 50 mg/L; Group 4, L-NAME (50 mg/L) plus mibefradil (50 mg. kg( 1). d(-1)); Group 5, L-NAME (50 mg/L) plus amlodipine (10 mg. kg(-1). d(-1)); Group 6 and 7, L-NAME (50 mg/L) for 3 weeks followed by mibefradil (50 mg. kg( 1). d(-1)) or amlodipine (10 mg. kg(-1). d(-1)), respectively, for the subsequent 3 weeks. Both the T- and L-channel calcium antagonists similarly reduced mean arterial pressure and total peripheral resistance index. These changes were associated with significant decreases in afferent and efferent glomerular arteriolar resistances and the ultrafiltration coefficient (P<0.01). Furthermore, the histopathological glomerular and arterial injury scores and urinary protein excretion were also significantly improved (P<0.01), and left ventricular and aortic masses were significantly diminished in all treated groups. Both drugs, mibefradil and amlodipine, had effects of increasing the single-nephron glomerular filtration ratio (SNGFR), and single-nephron plasma flow (SNPF), and of reducing glomerular afferent arteriolar resistance and urinary protein excretion. Thus, the T-type (mibefradil) and L-type (amlodipine) calcium antagonists each prevented and reversed the pathophysiological alterations of L NAME-exacerbated hypertensive nephrosclerosis in SHR. The T-type calcium antagonist (mibefradil) seemed to have been more effective than the L-type amlodipine antagonist and it produced a greater reduction in afferent arteriolar resistance while preserving SNGFR. PMID- 10454454 TI - Prevention of renal damage by angiotensin II blockade, accompanied by increased renal hepatocyte growth factor in experimental hypertensive rats. AB - Hepatocyte growth factor (HGF) is a unique growth factor that has many protective functions against renal damage. Our previous study demonstrated that HGF stimulated the growth of endothelial and epithelial cells without the replication of mesangial cells. Moreover, angiotensin (Ang) II significantly decreased local HGF production in mesangial cells. Therefore, we examined the effects of Ang II blockade on renal HGF expression and renal damage in experimental hypertensive rats. An angiotensin-converting enzyme inhibitor (cilazapril; 10 mg. kg(-1). d( 1)), an Ang II type 1 receptor antagonist (E-4177; 30 mg. kg(-1). d(-1)), hydralazine (8 mg. kg(-1). d(-1)), and vehicle were administered to 16-week-old stroke-prone spontaneously hypertensive rats (SHR-SP) for 3 weeks. Renal damage was evaluated with a computer analysis system, and renal HGF mRNA was measured by Northern blot analysis. Blood pressure of SHR-SP was significantly decreased by all drug treatments compared with vehicle. Moreover, cilazapril, E-4177, and hydralazine significantly decreased the thickening and necrosis of blood vessels compared with vehicle. Similarly, degeneration and necrosis of glomeruli were also markedly improved by cilazapril and E-4177 (P<0.01). We next examined the effects of Ang II blockade on renal HGF expression in SHR-SP. Renal HGF mRNA was markedly decreased in SHR-SP compared with Wistar-Kyoto rats, although Ang II blockade by cilazapril and E-4177 but not hydralazine significantly increased renal HGF mRNA in SHR-SP. Ang II blockade significantly increased renal HGF (a protective growth factor for tubular epithelial cells); thus, we examined tubular histological appearance. Degeneration and necrosis of tubules were significantly improved by cilazapril and E-4177 treatment (P<0.01). In addition, cell infiltration into the glomeruli and hemorrhage were also significantly reduced in SHR-SP treated with cilazapril or E-4177. The present data demonstrated the prevention of renal damage by Ang II blockade in SHR-SP, which was accompanied by a significant increase in renal HGF mRNA. Given the strong mitogenic activity and antiapoptotic actions of HGF on endothelial and epithelial cells, we believe that increased local HGF production by the blockade of Ang II may improve renal function in hypertension. PMID- 10454455 TI - Comparative effect of angiotensin-converting enzyme inhibition and angiotensin II type 1 receptor antagonism on plasma fibrinolytic balance in humans. AB - Angiotensin-converting enzyme (ACE) inhibition significantly decreases plasminogen activator inhibitor-1 (PAI-1) without altering tissue plasminogen activator (tPA) during activation of the renin-angiotensin-aldosterone system in humans. Because ACE inhibitors and angiotensin II type 1 (AT(1)) receptor antagonists differ in their effects on angiotensin II formation and bradykinin degradation, the present study compared the effect of equivalent hypotensive doses of an ACE inhibitor and AT(1) antagonist on fibrinolytic balance. Plasma PAI-1 antigen, tPA antigen, plasma renin activity, and aldosterone were measured in 25 normotensive subjects (19 white, 6 black; 14 men, 11 women; mean age 38.5+/ 1.8 years; mean body mass index 25.3+/-0.7 kg/m(2)) during low salt intake alone (10 mmol Na/d), low salt intake + quinapril (40 mg PO bid), and low salt intake + losartan (50 mg PO bid). Compared with low salt alone (systolic blood pressure [BP] 118.8+/-2.2 mm Hg), both quinapril (106.3+/-2.5 mm Hg, P<0.001) and losartan (105.4+/-2. 8 mm Hg, P<0.001) reduced BP. No statistical difference was found between quinapril and losartan in their BP lowering effect. Losartan (P=0.009), but not quinapril, lowered heart rate. Both drugs significantly lowered aldosterone (P<0.001 versus low salt alone for each); however, this effect was significantly greater for quinapril than for losartan (P<0.001 for quinapril versus losartan). Treatment with quinapril, but not with losartan, was associated with a decrease in both PAI-1 antigen (P=0.03) and activity (P=0.018). PAI-1 activity was lower during treatment with quinapril than with losartan (P=0.015). The average PAI-1 antigen concentration was 13. 0+/-2.0 ng/mL during low salt alone, 10.5+/-1.6 ng/mL during quinapril treatment, and 12.3+/-2.1 ng/mL during losartan treatment. In contrast, plasma tPA antigen concentrations were reduced during treatment with losartan (P=0.03) but not with quinapril. This study provides the first evidence that ACE inhibitors and AT(1) antagonists differ in their effects on fibrinolytic balance under conditions of activation of the renin angiotensin-aldosterone system. Further studies are needed to address the mechanism for the contrasting effects of these 2 classes of drugs on fibrinolysis and to define the clinical significance of these differences. PMID- 10454456 TI - Late treatment with ramipril increases survival in old spontaneously hypertensive rats. AB - Spontaneously hypertensive rats (SHR) begin to die from cardiovascular complications at approximately 15 months of age. We tested whether chronic ACE inhibitor treatment would extend the lifespan of such old animals. We also studied cardiac hypertrophy and function, endothelial function and expression, and activity of NO synthase (eNOS). One hundred 15-month-old SHR were randomized into 3 groups, control (n=10), placebo-treated (n=45), and ramipril-treated with an antihypertensive dose of 1 mg. kg(-1). d(-1) in drinking water (n=45). Ex vivo experiments were performed after 15 months (control) and 21 months, when approximately 80% of the placebo group had died. Late treatment with ramipril significantly extended lifespan of the animals from 21 to 30 months. Fully established cardiac hypertrophy, observed in placebo-treated animals and in controls, was significantly reversed by ramipril treatment. In isolated working hearts, a significantly improved function associated with increased cardiac eNOS expression was seen versus placebo and control hearts. Endothelial dysfunction in isolated aortic rings from control and placebo-treated SHR was significantly improved by ACE inhibition and associated with enhanced NO release. Late treatment of SHR with the ACE inhibitor ramipril extended lifespan from 21 to 30 months, which is comparable to the lifespan of untreated normotensive Wistar Kyoto rats. This lifespan extension, probably due to blood pressure reduction, correlated with increased eNOS expression and activity followed by a regression of left ventricular hypertrophy and cardiac and vascular dysfunction. PMID- 10454457 TI - Angiotensin-(1-7) is a modulator of the human renin-angiotensin system. AB - The renin-angiotensin system is important for cardiovascular homeostasis. Currently, therapies for different cardiovascular diseases are based on inhibition of angiotensin-converting enzyme (ACE) or angiotensin II receptor blockade. Inhibition of ACE blocks metabolism of angiotensin-(1-7) to angiotensin (1-5) and can lead to elevation of angiotensin-(1-7) levels in plasma and tissue. In animal models, angiotensin-(1-7) itself causes or enhances vasodilation and inhibits vascular contractions to angiotensin II. The function of angiotensin-(1 5) is unknown. We investigated whether angiotensin-(1-7) and angiotensin-(1-5) inhibit ACE or antagonize angiotensin-induced vasoconstrictions in humans. ACE activity in plasma and atrial tissue was inhibited by angiotensin-(1-7) up to 100%, with an IC(50) of 3.0 and 4.0 micromol/L, respectively. In human internal mammary arteries, contractions induced by angiotensin I and II and the non-ACE specific substrate [Pro(11),D-Ala(12)]-angiotensin I were antagonized by angiotensin-(1-7) (10(-5) mol/L) in a noncompetitive way, with a 60% inhibition of the maximal response to angiotensin II. Contractions to ACE-specific substrate [Pro(10)]-angiotensin I were also inhibited, an effect only partly accounted for by antagonism of angiotensin II. Angiotensin-(1-5) inhibited plasma ACE activity with a potency equal to that of angiotensin I but had no effect on arterial contractions. In conclusion, angiotensin-(1-7) blocks angiotensin II-induced vasoconstriction and inhibits ACE in human cardiovascular tissues. Angiotensin-(1 5) only inhibits ACE. These results show that angiotensin-(1-7) may be an important modulator of the human renin-angiotensin system. PMID- 10454458 TI - Activation of the brain angiotensin system by in vivo human angiotensin converting enzyme gene transfer in rats. AB - The possibility of the brain-specific expression of a component of the renin angiotensin system was evaluated in the present study. We used the hemagglutinating virus of Japan-liposome complex to transfect human angiotensin converting enzyme (ACE) cDNA, driven by the cytomegalovirus enhancer and beta actin promoter, into the lateral cerebroventricle of male Sprague-Dawley rats. We evaluated the time course of hemodynamics, the tissue levels of angiotensin (Ang) II and vasopressin, and ACE activity. Intracerebroventricular transfection of the human ACE gene increased both blood pressure and heart rate. Transfected rats exhibited higher concentrations of brain Ang II and increased brain ACE activity. This activation of the brain angiotensin system was accompanied by increased vasopressin production. The increases in blood pressure and heart rate were abolished by intracerebroventricular administration of an ACE inhibitor or Ang II type 1 receptor antagonist. The expression of the transgene was widely distributed in the periventricular cell layer, the cortex, the hypothalamic nuclei, and the brain stem. Expression in the neuronal cells persisted for up to 14 days. Thus, this hemagglutinating virus of Japan-liposome method is a highly efficient system for gene delivery and is extremely useful for functional gene transfection. This novel hypertensive model may enable characterization of the functions of the renin-angiotensin system in the brain and determination of its role in the pathogenesis of hypertension. PMID- 10454459 TI - Renin activity and blood pressure in response to chronic episodic hypoxia. AB - Previous studies in several strains of rats have demonstrated that 35 days of recurrent episodic hypoxia (EH) (7 hours per day), with a fractional concentration of inspired oxygen that produces desaturation equivalent to the recurrent hypoxemia of sleep apnea, results in an 8 to 13 mm Hg persistent increase in diurnal systemic blood pressure (BP). Carotid chemoreceptors and the sympathetic nervous system have been shown to be necessary for development of this BP increase. Both renal artery denervation and adrenal demedullation block the BP response to chronic EH. The present study was undertaken to define further the role of the kidneys and the renin-angiotensin system in this BP increase. Separate groups of male Sprague-Dawley rats had either (1) bilateral renal artery denervation with EH, (2) sham surgery with EH, (3) sham surgery with sham EH (compressed air), (4) EH with losartan, (5) unhandled with losartan, or (6) unhandled. The experimental period lasted 35 days. Both renal-artery denervated and losartan-treated animals showed no BP change or a lowering of BP in response to EH, whereas the sham-operated EH animals showed a progressive, sustained increase in resting room air BP. BP remained at basal levels or fell in unhandled and unhandled losartan-treated animals. Plasma renin activity was elevated 4-fold versus basal levels in EH animals with renal nerves intact but remained at baseline levels in denervated animals. At the end of the experiment, renal tissue catecholamines confirmed renal denervation in those animals. In conclusion, EH causes a progressive increase in BP, mediated in part through renal sympathetic nerve activity that acts to increase renin-angiotensin system activity through angiotensin II type 1 receptors. PMID- 10454461 TI - Hypertension and hormone-related neoplasms in women. AB - The relation between hypertension and the risk of selected hormone-related neoplasms in women was investigated in a network of case-control studies conducted in Italy during 1983-1996. Cases were women younger than 75 years with histologically confirmed cancer of the breast (n=3406), endometrium (n=745), ovary (n=970), and thyroid (n=145). Controls were 3054 women admitted in the same geographic area for acute, nonneoplastic, non-hormone-related diseases. Odds ratios (ORs) of treated hypertension were computed after allowance for sociodemographic factors, smoking habits, alcohol consumption, parity, menopausal status, and body mass index (BMI) by means of unconditional logistic regression. The ORs were 1.2 (95% CI, 1.1 to 1.4) for breast cancer and 1.6 (95% CI, 1.3 to 1.9) for endometrial cancer, and the elevated ORs persisted after >/=5 years since diagnosis of hypertension. No significant association was observed for ovarian and thyroid cancer. For breast cancer, the association was apparently stronger at age 55 years or over and consequently after menopause. No appreciable effect modification was evident for endometrial cancer. Allowance for BMI did not explain the association of postmenopausal breast cancer and endometrial cancer with hypertension. The OR of postmenopausal breast cancer was 1.5 (95% CI, 1.1 to 2.0) in hypertensive women with BMI >/=30 kg/m(2) compared with normotensive women with BMI <25 kg/m(2). The corresponding figure for all endometrial cancers was 4.9 (95% CI, 3. 4 to 6.9). Even in the absence of a clear understanding of biological mechanisms, the definition of a role of hypertension on female hormone related cancers can have relevant implications on individual risk assessment. PMID- 10454460 TI - Levels of mineralocorticoids in whites and blacks. AB - Blacks appear, on average, to retain more Na than whites. A higher production rate of mineralocorticoids could explain the greater Na retention in blacks. Although production of aldosterone has been shown to be lower in blacks, the level of another mineralocorticoid may be increased. Plasma levels of deoxycorticosterone and cortisol were measured in young whites (n=23; age=16.4+/ 3.1[SD] years) and young blacks (n=25; age=13.8+/-1.3 years). Blacks had lower plasma levels of renin activity and aldosterone and lower urinary aldosterone excretion rates; thus, they appeared to be representative of blacks that retain additional Na. Plasma deoxycorticosterone levels were lower in blacks than in whites both at baseline (247+/-161 versus 381+/-270 pmol/L, P=0.048) and after stimulation with adrenocorticotropic hormone (822+/-294 versus 1127+/-628 pmol/L at 30 minutes, P=0.047; 925+/-366 versus 1440+/-834 pmol/L at 60 minutes, P=0.013). Cortisol levels were also lower in blacks at baseline (P=0.014) but were not significantly different from levels in whites after stimulation with adrenocorticotropic hormone. In a larger cohort of 407 whites (age=12.0+/-2.9 years) and 247 blacks (age=12.9+/-3.1 years), 18-hydroxycortisol excretion rates were also lower in blacks (P=0. 021). In conclusion, increased Na retention in blacks does not appear to be secondary to increased production of either aldosterone, deoxycorticosterone, cortisol, or 18-hydroxycortisol. A primary renal mechanism may mediate the increase in Na reabsorption in blacks. PMID- 10454462 TI - 5-Hydroxytryptamine(1F) receptors do not participate in vasoconstriction: lack of vasoconstriction to LY344864, a selective serotonin(1F) receptor agonist in rabbit saphenous vein. AB - Recently, several novel approaches to the treatment of migraine have been advanced, including selective 5-hydroxytryptamine (or serotonin) 1B/1D (5 HT(1B/1D)) receptor agonists such as sumatriptan and 5-HT(1F) receptor agonists such as LY344864. Many 5-HT(1B/1D) receptor agonists have been identified based on their ability to produce cerebral vascular contraction, whereas LY344864 was identified as an inhibitor of trigeminal nerve-mediated dural extravasation. In our study, several triptan derivatives were compared with LY344864 for their ability to contract the rabbit saphenous vein, a tissue used in the preclinical identification of sumatriptan-related agonists. Sumatriptan, zolmitriptan, rizatriptan, and naratriptan all contracted the rabbit saphenous vein from baseline tone, whereas LY344864 in concentrations up to 10(-4) M did not contract the rabbit saphenous vein. Furthermore, vascular contractions to sumatriptan were markedly augmented in the presence of prostaglandin F(2alpha) (PGF(2alpha)). However, even in the presence of PGF(2alpha) (3 x 10(-7) M), LY344864 did not contract the rabbit saphenous vein in concentrations well in excess of its 5 HT(1F) receptor affinity (pK(i) = 8.2). Only when concentrations exceeded those likely to activate 5-HT(1B) and 5-HT(1D) receptors (>10(-5) M) did modest contractile responses occur in the presence of PGF(2alpha). Use of these serotonergic agonists revealed a significant correlation between the contractile potency in the rabbit saphenous vein and the affinities of these agonists at 5 HT(1B) and 5-HT(1D) receptors, although contractile agonist potencies were not quantitatively similar to 5-HT(1B) or 5-HT(1D) receptor affinities. In contrast, no significant correlation existed between the contractile potencies of these serotonergic agonists in the rabbit saphenous vein and their affinity at 5-HT(1F) receptors. These data support the contention that activation of 5-HT(1F) receptors will not result in vascular contractile effects. PMID- 10454463 TI - Cationic modulation of human dopamine transporter: dopamine uptake and inhibition of uptake. AB - Effects of cations on dopamine (DA) uptake into cells expressing the human dopamine transporter and on inhibition of DA uptake by various substrates and inhibitors were investigated by using rotating disk electrode voltammetry. The Na(+) dependence of DA uptake varied with Na(+) substitutes, hyperbolic with Li(+), almost linear at 1 microM DA but hyperbolic at 8 microM DA with choline, and sigmoidal with K(+). With Na(+) substituted by Li(+), K([DA]) decreased and V(app) remained constant with increasing [Na(+)], whereas K([Na+]) decreased and V(app) increased with increasing [DA], suggesting an ordered sequence with Na(+) binding before DA. Similar trends for the Na(+)-DA interactions were observed in the presence of cocaine. Cocaine inhibited DA uptake solely by increasing K([DA]), with its K(i) not significantly different at 55 and 155 mM [Na(+)], whereas it inhibited Na(+) stimulation by reducing V(app) more than K([Na+]) at 1 microM DA, and V(app) only and less potently at 8 microM DA. Thus, cocaine may compete with DA, not with Na(+), for the transporter, and might not follow a strictly ordered reaction with Na(+). With Na(+) substituted by K(+), K([DA]) or K([Na+]) became insensitive to Na(+) or DA. K(+) impaired the DA uptake mainly by reducing V(app,) but affected cocaine inhibition by elevating K(i). Despite their different patterns for inhibiting DA uptake, nontransportable inhibitors cocaine, methylphenidate, mazindol, and 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3 phenyl-2-propyl)piperazi ne (GBR12909) showed similarly modest Na(+) dependence in their K(i) values. In contrast, substrates DA, m-tyramine, and amphetamine displayed a similarly stronger Na(+) requirement for their apparent affinities. PMID- 10454464 TI - Glutathione-dependent metabolism of cis-3-(9H-purin-6-ylthio)acrylic acid to yield the chemotherapeutic drug 6-mercaptopurine: evidence for two distinct mechanisms in rats. AB - cis-3-(9H-Purin-6-ylthio)acrylic acid (PTA) is a structural analog of azathioprine, a prodrug of the antitumor and immunosuppressive drug 6 mercaptopurine (6-MP). In this study, we examined the in vitro and in vivo metabolism of PTA in rats. Two metabolites of PTA, 6-MP and the major metabolite, S-(9H-purin-6-yl)glutathione (PG), were formed in a time- and GSH-dependent manner in vitro. Formation of 6-MP and PG occurred nonenzymatically, but 6-MP formation was enhanced 2- and 7-fold by the addition of liver and kidney homogenates, respectively. Purified rat liver glutathione S-transferases enhanced 6-MP formation from PTA by 1.8-fold, whereas human recombinant alpha, mu, and pi isozymes enhanced 6-MP formation by 1.7-, 1.3-, and 1.3-fold, respectively. In kidney homogenate incubations, PG accumulation was only observed during the first 15 min because of further metabolism by gamma-glutamyltranspeptidase, dipeptidase, and beta-lyase to yield 6-MP, as indicated by the use of the inhibitors acivicin and aminooxyacetic acid. Based on these results and other lines of evidence, two different GSH-dependent pathways are proposed for 6-MP formation: an indirect pathway involving PG formation and further metabolism to 6 MP, and a direct pathway in which PTA acts as a Michael acceptor. HPLC analyses of urine of rats treated i.p. with PTA (100 mg/kg) showed that 6-MP was formed in vivo and excreted in urine without apparent liver or kidney toxicity. Collectively, these studies show that PTA is metabolized to 6-MP both in vitro and in vivo and may therefore be a useful prodrug of 6-MP. PMID- 10454465 TI - Mechanism of intestinal absorption of an orally active beta-lactam prodrug: uptake and transport of carindacillin in Caco-2 cells. AB - Absorption characteristics of carindacillin (CIPC) were investigated using Caco-2 cells, and the results were compared with those of its parent drug, carbenicillin (CBPC). Uptake of CBPC was not affected by the metabolic inhibitor or the change in extracellular pH. CBPC appeared to be taken up into Caco-2 cells by passive diffusion. In contrast, the uptake of CIPC was greater at lower extracellular pH and was inhibited in the presence of carbonyl cyanide p-(trifluoromethoxy)phenyl hydrazone, a protonophore. Also, transport of CIPC through Caco-2 cell monolayer was energy and temperature dependent. Moreover, the uptake and transport of CIPC were significantly inhibited in the presence of various monocarboxylic acids, which are the substrates of the monocarboxylic acid transport system(s), whereas the substrates of the oligopeptide transporter had no effect on the uptake or transport of CIPC. These results suggested that the absorption of CIPC may be mediated by the monocarboxylic acid transport system(s), not by the oligopeptide transporter. Furthermore, the uptake and transport of CIPC were approximately 40 fold greater than those of CBPC. Therefore, it is likely that the participation of a carrier-mediated transport in the absorption of CIPC may significantly contribute to the improved absorption of the prodrug over the parent drug. PMID- 10454467 TI - Effects of potassium channel blockers on CO2-induced slowly adapting pulmonary stretch receptor inhibition. AB - In anesthetized, artificially ventilated rabbits with vagus nerve section, inhalation of CO(2) gas mixtures (tracheal CO(2) concentration ranging from 8.0 to 10.2%) for 60 s decreased slowly adapting pulmonary stretch receptor (SAR) activity during both inflation and deflation. The magnitude of decreased receptor activity during deflation had a more pronounced effect than that seen during inflation. CO(2) inhalation did not cause any significant change in tracheal pressure (P(T)) as an index of bronchomotor tone. Intravenous administration of 4 aminopyridine (0. 7 and 2.0 mg/kg i.v.), a K(+) channel blocker, which dose dependently increased SAR activity during deflation and had no effect on P(T), abolished or attenuated the decrease in SAR activities induced by CO(2) inhalation in a dose-dependent manner. The K(+) channel blocker tetraethylammonium (2.0 and 6.0 mg/kg i.v.) that did not significantly alter either basal SAR discharge or P(T) had no effect on the inhibitory responses of receptor activity to CO(2) inhalation. These results suggest that the inhibitory mechanism of CO(2) inhalation on SARs may be involved in the activation of 4 aminopyridine-sensitive K(+) channels in the nerve terminals of SARs. PMID- 10454466 TI - Pharmacological studies of the acute and chronic effects of (+)-3, 4 methylenedioxymethamphetamine on locomotor activity: role of 5 hydroxytryptamine(1A) and 5-hydroxytryptamine(1B/1D) receptors. AB - The 5-hydroxytryptamine(1B/1D) (5-HT(1B/1D)) antagonist 2'-methyl-4'-(5-methyl [1,2,4]oxadiazol-3-yl)-biphenyl-4-carboxyli c acid [4-methoxy-3-(4-methyl piperazin-1-yl)-phenyl]-amide (GR 127935) and 5-HT(1A) antagonist N-(2-(4-(2 methoxyphenyl)-1-piperazinyl)ethyl)-N-(2-pyridinyl)cyclohe xanecarboxamide (WAY 100635) were used to assess whether hyperactivity induced by 3 mg/kg (+)-3, 4 methylenedioxymethamphetamine [(+)-MDMA] is mediated by 5-HT(1B/1D) and/or 5 HT(1A) receptors. Activity in the periphery and center of an open field as well as rearing activity were measured in photobeam monitors. (+)-MDMA-induced peripheral and central activities were blocked by GR 127935 (0.3, 0.625, 1.25, and 2.5 mg/kg); central hyperactivity was blocked by 0.1, 0.3, and 0.625 mg/kg GR 127935. WAY 100635 (0.5-2 mg/kg) had little effect on (+)-MDMA-induced activity except for an enhancement of central activity at one dose (0.5 mg/kg). Central activity induced by (+)-MDMA increased from day 1 to day 5 of treatment with (+) MDMA (3 mg/kg), whereas peripheral, central, and rearing activity significantly increased in (+)-MDMA-treated rats pretreated daily with GR 127935 (2.5 mg/kg). Withdrawal from (+)-MDMA, but not GR 127935 + (+)-MDMA, pretreatment was associated with heightened hyperactivity induced by the 5-HT(1B/1A) agonist RU 24969 (2 mg/kg i. p.); treatments were not associated with alterations in 5-HT and 5-hydroxyindoleacetic acid content or turnover in frontal cortex. These data support a role for 5-HT(1B/1D) in mediating the acute hyperactivity evoked by (+) MDMA. The development of sensitization to (+)-MDMA was associated with supersensitivity to a 5-HT(1B/1A) agonist, suggesting that these receptors may contribute to sensitization. However, sensitization to (+)-MDMA developed even under conditions of 5-HT(1B/1D) receptor blockade, which is somewhat counter to this speculation. Perhaps, under circumstances of continued 5-HT(1B/1D) blockade, other mechanisms (e.g., dopamine) predominate in the progressive enhancement of behavior with repeated (+)-MDMA treatment. PMID- 10454469 TI - Native gamma-aminobutyric acid type A receptors from rat hippocampus, containing both alpha 1 and alpha 5 subunits, exhibit a single benzodiazepine binding site with alpha 5 pharmacological properties. AB - Evidences indicate the existence of two homologous and/or heterologous alpha subunits coassembled in a single gamma-aminobutyric acid type A (GABA(A)) receptor. However, it is unknown whether both or only one of the coassembled alpha subunits display benzodiazepine binding sites. Thus, we have investigated the association between alpha1 and alpha5 subunits and the pharmacological properties of these GABA(A) receptors from rat hippocampus. The association between alpha1 and alpha5 subunits was demonstrated by immunoblot of the anti alpha1 or -alpha5 immunoaffinity-purified receptors and by double immunopurification by anti-alpha1 and -alpha5 columns in series. The benzodiazepine binding properties of the immunoprecipitated receptors indicated the existence of pharmacologically active and inactive alpha subunits. The anti alpha5 immunoprecipitated receptors displayed exclusively low-affinity binding sites for both Cl218,872 (K(i) = 0.81 +/- 0.15 microM) and zolpidem (K(i) = 5.0 +/- 3.0 microM), in spite of the association between alpha1 and alpha5 subunits. The anti-alpha1 immunoprecipitated receptors displayed both high- and low affinity binding sites for both ligands (K(i)s = 47.5 +/- 5.2 nM and 0.7 +/- 0.06 microM for Cl218,872 and 25.0 +/- 7.0 nM, 415 +/- 200 nM and 9. 3 +/- 3.0 microM for zolpidem). Therefore, the alpha5 subunit, when coassembled with alpha1 subunit, should be pharmacologically predominant. This hypothesis was probed by immunoprecipitation of the photoaffinity-labeled receptors and by anti-alpha1 and -alpha5 double immunopurified receptors. The alpha1-alpha5 double immunopurified receptors displayed a single low-affinity binding site (K(i) = 908 +/- 105 nM) for Cl218,872, undetectable [(3)H]zolpidem binding activity, and similar [(3)H]flumazenil and [(3)H]L-655,708 binding activity (0.10 +/- 0.01 and 0.09 +/- 0.02 pmol/20 microliters of anti-alpha5 immunobeads, respectively). Thus, the native GABA(A) receptors containing alpha1 and alpha5 subunits have only one alpha subunit pharmacologically active displaying alpha5 binding properties. PMID- 10454470 TI - Ruthenium red-mediated inhibition of large-conductance Ca2+-activated K+ channels in rat pituitary GH3 cells. AB - The ionic mechanism of actions of ruthenium red was examined in rat anterior pituitary GH(3) cells. In whole-cell recording experiments, ruthenium red reversibly caused an inhibition of Ca(2+)-activated K(+) current [I(K(Ca))] in a dose-dependent manner. The IC(50) value of ruthenium red-induced inhibition of I(K(Ca)) was 15 microM. Neither carbonyl cyanide m-chlorophenyl hydrazone (CCCP; 10 microM), an uncoupler of oxidative phosphorylation in mitochondria, nor cyclosporin A (200 nM), an inhibitor of the mitochondrial permeability transition pore, affected the amplitude of I(K(Ca)). In inside-out configuration, application of ruthenium red (50 microM) into the bath medium did not change single-channel conductance but significantly suppressed the activity of large conductance Ca(2+)-activated K(+) channel (BK(Ca)) channels. The ruthenium red induced decrease in the channel activity of BK(Ca) channels was reversed by an increase in intracellular Ca(2+) concentration. Ruthenium red also shifted the activation curve of BK(Ca) channels to positive membrane potentials. The change in the kinetic behavior of BK(Ca) channels caused by ruthenium red in these cells is due to a decrease in mean open time and an increase in mean closed time. Ruthenium red (50 microM) did not affect the amplitude of voltage-dependent K(+) current but produced a significant reduction of voltage-dependent L-type Ca(2+) current. These results indicate that ruthenium red can directly suppress the activity of BK(Ca) channels in GH(3) cells. This effect is independent on the inhibition of Ca(2+) release from internal stores or mitochondria. PMID- 10454468 TI - Coupling efficiencies of beta 1- and beta 2-adrenergic receptors expressed alone or together in transfected GH3 pituitary cells. AB - The relationship between rat beta(1)- and beta(2)-adrenergic receptors (ARs) and cyclic AMP (cAMP) responses was examined by inducible expression of each subtype in transfected GH(3) pituitary cells. Increasing expression of beta(1)- or beta(2)-ARs in stably transfected subclones increased basal cAMP, increased the potency of isoproterenol in stimulating cAMP formation, but did not change the maximal response. A linear relationship was observed between log B(max) and -log EC(50) for isoproterenol, with no significant differences between beta(1)- and beta(2)-ARs. When both subtypes were coexpressed at different densities and ratios, pharmacological analysis showed that both selective and nonselective agonists exerted their effects at least partially through both subtypes. Either subtype alone activated a maximal response when the other subtype was blocked, indicating a complete redundancy in coupling. Agonists could activate responses through either subtype, with responses mediated primarily through the subtype where the agonist was most potent. The nonselective agonist isoproterenol had similar potencies for activating both subtypes; however, the density and ratio of subtypes affected the relative potencies of the selective agonists norepinephrine and zinterol. We conclude that beta(1)- and beta(2)-ARs have similar coupling efficiencies in GH(3) cells, these efficiencies are not altered by coexpression of another subtype, they couple redundantly to cAMP formation, and the relative densities of beta(1)- and beta(2)-ARs control the potencies of selective agonists. PMID- 10454471 TI - Non-cyclic AMP-dependent, positive inotropic cyclodepsipeptides with negative chronotropy. AB - The effects of natural cyclodepsipeptides (CDPs) on isolated rat cardiac tissue preparations were examined in vitro. Destruxin A, destruxin B (DB), roseotoxin B (RB), and roseocardin (RC), a novel CDP, each caused a concentration-dependent increase in the contraction force of the right atrium and the papillary and trabecular muscles of the right ventricle at 0.6 to 600 microM. RB, destruxin A, and DB did not affect the half-decay time of relaxation of the papillary muscles, but RC slightly prolonged it, although to a much lesser extent than BA 41899, a calcium sensitizer. This inotropic effect is accompanied by a prolongation of the automatic atrial contraction intervals. The RB-induced increase in the contraction force of papillary muscle was not affected by phentolamine, propranolol, pyrilamine, or cimetidine. RB- and RC-induced increases in the contraction force of papillary muscles were not affected by 3-isobutyl-1 methylxanthine or carbachol. Neither peptide changed the cyclic AMP levels in trabecular muscles. Neither RB nor RC affected the activity of Na(+),K(+)-ATPase from rat kidney. Neither RB, RC, nor DB affected the resting membrane potential or the apparent input resistance of papillary muscles. These results suggest that these CDPs produce both non-cyclic AMP-dependent positive inotropic and negative chronotropic effects. PMID- 10454472 TI - Z-350, a novel compound with alpha 1-adrenoceptor antagonistic and steroid 5 alpha-reductase inhibitory actions: pharmacological properties in vivo. AB - The alpha(1)-adrenoceptor-antagonistic and steroid 5alpha-reductase-inhibitory actions of Z-350 [(S)-4-{3-{4-{1-(4-methylphenyl)-3-[4-(2 methoxyphenyl)piperazine-1-y l]propoxy}benzoyl}indole-1-yl}butyric acid hydrochloride] were investigated in rabbits and rats in vivo. Z-350 (1-30 mg/kg), administered intraduodenally, dose-dependently inhibited phenylephrine-induced increases in prostatic urethral pressure with an ED(50) value of 3.8 mg/kg in anesthetized male rabbits, whereas the effects on mean blood pressure and orthostatic hypotensive response were weaker when compared with other alpha(1) adrenoceptor antagonists, tamsulosin and prazosin. Z-350 (1-10 mg/kg p.o.) dose dependently inhibited the prostatic steroid 5alpha-reductase activity in rats with an ED(50) value of 2.8 mg/kg. The daily oral administration of Z-350, at >==10 mg/kg for 7 days, significantly reduced the prostatic growth induced by testosterone in castrated rats, with no effect on dihydrotestosterone-induced prostatic growth. These results indicate that Z-350 exhibited alpha(1) adrenoceptor-antagonistic and 5alpha-reductase inhibitory actions at almost equal doses in vivo, and was expected to improve the bladder outlet obstruction associated with benign prostatic hyperplasia with smaller cardiovascular adverse effect. PMID- 10454474 TI - Stimulatory and inhibitory properties of aminoglycoside antibiotics at N-methyl-D aspartate receptors. AB - The effects of aminoglycoside antibiotics on N-methyl-D-aspartate (NMDA) receptors were studied using voltage-clamp recording of recombinant NMDA receptors expressed in Xenopus oocytes. A number of aminoglycosides were found to potentiate macroscopic currents at heteromeric NR1A/NR2B receptors, but not at NR1A/NR2A, NR1A/NR2C, NR1A/NR2D, or NR1B/NR2B receptors. The degree of potentiation had a rank order neomycin B > paromomycin > gentamicin C > geneticin > kanamycin A > streptomycin. Potentiation was not seen with kasugamycin and spectinomycin. The degree of stimulation paralleled the number of the amino groups in the aminoglycosides. The stimulatory effects of aminoglycosides were more pronounced at subsaturating concentrations of glycine and at acidic pH, similar to the stimulatory effects of spermine. We measured the effects of aminoglycosides at mutant NMDA receptors to determine which amino acid residues in NMDA receptor subunits are involved in stimulation. Mutations that reduced or abolished spermine stimulation also reduced stimulation by aminoglycosides. Several aminoglycosides produced a weak voltage-dependent block of NMDA receptors, but the degree of inhibition did not appear to correlate with the number of amino groups in the molecule. The results suggest that aminoglycosides having more than three amino groups have stimulatory effects that are mediated through the spermine-binding site on NMDA receptors. PMID- 10454473 TI - Further pharmacological characterization of bradykinin B1 receptor up-regulation in human umbilical vein. AB - Previous reports have provided evidence to support the view that the de novo synthesis of bradykinin (BK) B(1) receptor is involved in the induction of vascular responses in human umbilical vein (HUV). In the present study, we evaluated different pharmacological tools to further analyze this up-regulation process in HUV. Concentration-response curves to des-Arg(9)-BK, a selective BK B(1) receptor agonist, were performed after 5 h of incubation. Tumor necrosis factor-alpha potentiated BK B(1) receptor responses at 5 h without modifying the maximal response to des-Arg(9)-BK. Pyrrolidine dithiocarbamate, an inhibitor of nuclear factor-kappaB activation, produced a concentration-dependent decrease of the BK B(1) receptor sensitization. When tissues were continuously exposed to actinomycin D, a transcription inhibitor, or cycloheximide, a protein synthesis inhibitor, concentration-response curves to des-Arg(9)-BK were markedly diminished. On the other hand, transitory exposure to cycloheximide allowed the full recovery of BK B(1) receptor-sensitized responses at 5 h. Finally, continuous incubation with the N-linked glycosylation inhibitor, tunicamycin, almost completely abolished des-Arg(9)-BK-mediated responses. In summary, this sensitization process is potentiated by tumor necrosis factor-alpha and is selectively inhibited by pyrrolidine dithiocarbamate, suggesting that BK B(1) receptor up-regulation in HUV involves nuclear factor-kappaB activation. The effects of actinomycin D and tunicamycin provide evidence that the de novo synthesis of a transmembrane glycoprotein has an obligatory role in the BK B(1) up-regulation. The reversion of the cycloheximide effect on BK B(1) response indicates that the time necessary for synthesis, trafficking, and functional membrane expression of this receptor would be less than 1 h. PMID- 10454475 TI - Differing effects of N-methyl-D-aspartate receptor subtype selective antagonists on dyskinesias in levodopa-treated 1-methyl-4-phenyl-tetrahydropyridine monkeys. AB - The antiparkinsonian and antidyskinetic profile of two N-methyl-D-aspartate (NMDA) receptor antagonists, a competitive antagonist, (R)-4-oxo-5 phosphononorvaline (MDL 100,453), and a novel noncompetitive allosteric site antagonist, 4-hydroxy-N-[2-(4-hydroxyphenoxy)ethyl]-4-(4-methylbenzyl)piper idi ne (Co 101244/PD 174494), was assessed in six levodopa-treated 1-methyl-4-phenyl tetrahydropyridine-lesioned parkinsonian monkeys. The effects on motor function of these two drugs, alone and in combination with levodopa, were then correlated with NMDA subtype selectivity and apparent affinity for four diheteromeric NMDA receptor subunit combinations expressed in Xenopus oocytes. MDL 100, 453 (300 mg/kg s.c.) by itself increased global motor activity (p =. 0005 versus vehicle) and administered 15 min after a low dose of levodopa/benserazide s.c., MDL 100,453 (50, 300 mg/kg s.c.) showed dose-dependent potentiation of antiparkinsonian responses and also produced dyskinesias. Following injection of a fully effective dose of levodopa, MDL 100,453 (300 mg/kg s.c.) also produced a 25% increase in mean dyskinesia score (p =.04). In contrast, Co 101244 did not change motor activity by itself and only showed a tendency to potentiate the antiparkinsonian response when given in combination with a low dose of levodopa, which did not attain statistical significance. However, with a high dose of levodopa, Co 101244 (0.1, 1 mg/kg s.c.) displayed antidyskinetic effects (67 and 71% reduction, respectively) while sparing levodopa motor benefit. In vitro, MDL 100,453 was an NMDA glutamate-site antagonist, with approximately 5- to 10-fold selectivity for the NR1A/NR2A subtype combination (K(b) = 0.6 microM) versus NR1A in combination with 2B, 2C, or 2D. In contrast, the allosteric site antagonist Co 101244 showed approximately 10,000-fold selectivity for the NR1A/NR2B (IC(50) = 0.026 microM) versus the other three subunit combinations tested. Taken together, the data suggest that the NR2 subunit selectivity profile of NMDA receptor antagonists can play an important role in predicting behavioral outcome and offer more evidence that NR2B-selective NMDA receptor antagonists may be useful agents in the treatment of Parkinson's disease. PMID- 10454476 TI - Ex vivo reversal of heparin-mediated cardioprotection by heparinase after ischemia and reperfusion. AB - Glycosaminoglycans, including heparin, have been demonstrated both in vitro and in vivo to protect the ischemic myocardium against reperfusion injury. In the present study, we sought to determine whether the cardioprotective effects of heparin administration could be reversed by the heparin-degrading enzyme heparinase. New Zealand white rabbits were pretreated with heparin (300 U/kg i.v.) or vehicle (saline). Two hours after treatment, hearts were removed, perfused on a Langendorff apparatus, and subjected to 25 min of global ischemia, followed by 45 min of reperfusion. Hemodynamic variables were obtained before ischemia (baseline) and every 10 min throughout the reperfusion period. Compared with vehicle-treated rabbits, the left ventricular end-diastolic and left ventricular developed pressures were improved significantly (p <.05) in the heparin-treated group. Ex vivo administration of heparinase (5 U/ml) immediately before the onset of global ischemia was associated with a reversal of the heparin mediated cardioprotection. The uptake of a radiolabeled antibody to the intracellular protein myosin and creatine kinase release were used to determine membrane integrity and discriminate between viable and nonviable myocardial tissue. The uptake of radiolabeled antimyosin antibody and release of creatine kinase after reperfusion were increased in heparin-pretreated hearts exposed to heparinase, indicating a loss of membrane integrity and increased myocyte injury. These results demonstrate that neutralization of heparin by heparinase promotes increased myocardial injury after reperfusion of the ischemic myocardium. PMID- 10454477 TI - Influence of hypovolemia on the pharmacokinetics and the electroencephalographic effect of etomidate in the rat. AB - The influence of hypovolemia (removal of 30% of the blood volume) on the pharmacokinetics and pharmacodynamics of etomidate was investigated in the rat. Chronically instrumented animals were randomly allocated to either a control (n = 9) or a hypovolemia (n = 9) group, and etomidate was infused (50 mg/kg/h) until isoelectric periods of 5 s or longer were observed in the electroencephalogram. The changes observed in the electroencephalogram were quantified using aperiodic analysis in the 2.5- to 7.5-Hz frequency band and used as a surrogate measure of hypnosis. The righting reflex was used as a clinical measure of hypnosis. The etomidate dose that had to be infused to reach the electroencephalographic endpoint was almost 40% lower (p <.01) in the hypovolemic animals than in the control animals. This difference could be attributed to a decrease in clearance ( 20%; p =.06) and distribution volume (-30%; p <.01) of etomidate. Protein binding was similar in both groups. To investigate changes in end organ sensitivity during hypovolemia, the electroencephalographic effect-versus-effect-site concentration relationship was studied. The effect-plasma concentration relationship was biphasic, exhibiting profound hysteresis in both hypovolemic and control animals. Semiparametric minimization of this hysteresis revealed similar equilibrium half-lives in both groups, and the biphasic effect-concentration relationship was characterized nonparametrically by descriptors. With these descriptors, a slightly increased potency of etomidate during hemorrhage was observed. The concentration at the return of righting reflex was 16% (p <.05) lower in the hypovolemic animals. In conclusion, an increased hypnotic effect of etomidate was observed during hypovolemia that is mainly attributed to pharmacokinetic changes. Our data also suggest a small increase in central nervous system sensitivity for etomidate in hypovolemic animals. PMID- 10454478 TI - Enhancement of osteogenesis in vitro and in vivo by a novel osteoblast differentiation promoting compound, TAK-778. AB - TAK-778 [(2R,4S)-(-)-N-(4-diethoxyphosphorylmethylphenyl)-1,2,4, 5-tetrahydro-4 methyl-7, 8-methylenedioxy-5-oxo-3-benzothiepin-2-carboxyamide; mw 505.53], a novel osteoblast differentiation promoting compound, was characterized in vitro and in vivo models. TAK-778 at doses of 10(-6) M and higher promoted potently bone-like nodule formation in the presence of dexamethasone in rat bone marrow stromal cell culture. This was accompanied by increases in cellular alkaline phosphatase activity, soluble collagen release, and osteocalcin secretion. Under the culture conditions, TAK-778 also stimulated the secretion of transforming growth factor-beta and insulin-like growth factor-I, indicating that TAK-778 may exert regulatory effects on osteoblast differentiation via autocrine/paracrine mechanisms. Furthermore, the in vivo osteogenic potential of TAK-778 was studied in bony defect and osteotomy animal models, using sustained release microcapsules consisted of a biodegradable polymer, poly (dl-lactic/glycolic) acid (PLGA). Single local injection of TAK-778/PLGA-microcapsules (PLGA-MC) (0.2-5 mg/site) to rat skull defects resulted in a dose-dependent increase in new bone area within the defects after 4 weeks. When the pellet containing TAK-778/PLGA-MC (4 mg/pellet) was packed into place to fill the tibial segmental defect in rabbit, this pellet induced osseous union within 2 months, whereas the placebo pellet did not. In addition, single local application of TAK-778/PLGA-MC (10 mg/site) to rabbit tibial osteotomy site enhanced callus formation accompanied by an increase in breaking force after 30 days. These results reveal for the first time that a nonendogenous chemical compound promotes potently osteogenesis in vitro and enhances new bone formation during skeletal regeneration and bone repair in vivo and should be useful for the stimulation of fracture healing. PMID- 10454479 TI - Manipulation of the tetrahydrocannabinol side chain delineates agonists, partial agonists, and antagonists. AB - Structure-activity relation studies have established that the alkyl side chain in tetrahydrocannabinol (THC) plays a crucial role in the activation of the cannabinoid receptor. Unfortunately, the flexible nature of this side chain has hampered efforts to elucidate the precise nature of the interaction of THC with its receptors. Therefore, a series of analogs with structurally restrained side chains of varying length was synthesized and evaluated for pharmacological potency in mice and for receptor affinity. The introduction of cis double bonds inserted rigid angles, whereas triple bonds developed regions of planarity. Receptor affinity for the acetylenic and saturated side chains were the same, whereas double bond substitution increased affinity 10-fold. Moreover, the relationship between receptor affinity and potency was 10-fold less than that of Delta(8)-THC in the case of some acetylenic derivatives, whereas changing the triple bond to a double bond restored the potency/affinity ratio. Additionally, an acetylene at C2-C3 in the octyl and nonyl side chains favored antinociception by as much as 70-fold. Surprisingly, several high-affinity acetylenic derivatives, especially those with cyano substitutions at the terminus of the side chain, were partial agonists or were inactive. Some of these low-efficacy, high-affinity ligands elicited antagonistic activity. The finding that manipulations of the side chain produces high- affinity ligands with either antagonist, partial agonist, or full agonist effects reveals a critical structural feature for receptor activation. PMID- 10454480 TI - Troglitazone inhibits bicarbonate secretion in rat and human duodenum. AB - Troglitazone is a new, orally effective antidiabetic agent that decreases plasma glucose in obese patients with non-insulin-dependent diabetes mellitus. Unfortunately, troglitazone also has a propensity to cause edema. This study was designed to determine how troglitazone affects intestinal ion transport and water absorption. Short circuit current (I(sc)) was measured in rat and human duodenal mucosa in Ussing chambers. Five minutes later, the serosal addition of troglitazone caused I(sc) to decrease gradually, and after 50 min, I(sc) reached the peak of decrease. EC(50) values and maximum response to I(sc) in rat and human mucosa were 8.4 and 8.7 microM and 8.56 +/- 1.0 and 8.00 +/- 2.0 microA/cm(2), respectively. In an HCO(3)(-)/CO(2)-free system, the decrease in I(sc) caused by troglitazone was 1.31 +/- 0.83 microA/cm(2). When 10 mM acetazolamide was preadministered, the small decrease in I(sc) evoked by troglitazone (20 microM) was 4.56 +/- 0.22 microA/cm(2), whereas the preadministration of 100 microM amiloride and 100 nM tetrodotoxin did not influence the decrease in I(sc) evoked by troglitazone. The serosal preadministration of 100 nM vasoactive intestinal peptide potently enhanced the decrease in I(sc) evoked by 20 microM troglitazone (21.1 +/- 1.63 microA/cm(2)). The cyclic AMP contents of rat duodenal mucosa incubated with and without troglitazone (20 microM) for 50 min were 3.2 +/- 0.25 and 5. 8 +/- 0.46 pmol/mg protein, respectively (P <.01). These results indicate that the ionic basis for the decrease in I(sc) that is induced by troglitazone may be inhibition of electrogenic bicarbonate secretion. The alteration of intestinal ion transport by troglitazone could cause edema. PMID- 10454481 TI - Pharmacological profile of ZD1611, a novel, orally active endothelin ETA receptor antagonist. AB - The endothelins (ETs), potent vasoconstrictor peptides, have been implicated in the pathogenesis of various cardiovascular disorders. In the present study, we describe the novel, potent, orally active, selective ET(A) receptor antagonist ZD1611 [3-(4-[3-(3-methoxy-5-methylpyrazin-2-ylsulfamoyl)-2-pyridyl ]phenyl)- 2,2 dimethylpropanoic acid]. ZD1611 competitively inhibited (125)I-labeled ET-1 binding at human cloned ET(A) and ET(B) receptors with pIC(50) values of 8.6 +/- 0.1 and 5.6 +/- 0.1, respectively, showing 1000-fold selectivity for the ET(A) receptor. ZD1611 caused a parallel rightward shift of the concentration response curve to ET-1 in the rat isolated aorta yielding a concentration of antagonist that caused a 2-fold rightward shift in the ET-1-response curve (pA(2)) of 7.5 +/ 0.3. When administered i. v. to anesthetized rats and dogs, ZD1611 caused dose related rightward shifts of partial dose-response curves to the precursor of ET 1, big ET-1. Threshold doses for significant antagonist activity were determined as 0.1 mg/kg and 0.3 mg/kg in the rat and dog, respectively. Importantly, ZD1611 was able to reverse an established big ET-1-induced pressor response in pithed rats in the presence of continuous big ET-1 infusion. Failure of ZD1611 to inhibit the BQ3020 (ET(B)-selective)-induced depressor response in pithed rats indicated a lack of activity at the endothelial ET(B) receptor. ZD1611 was orally active in the rat at 0.3 mg/kg and had a duration of action of more than 7 h, and, in the dog, a dose of 0.6 mg/kg p.o. was active for at least 6 h. In conclusion, these data demonstrate that ZD1611 is a potent and orally active, selective ET(A) receptor antagonist with a long duration of action which may be of therapeutic use. PMID- 10454482 TI - Discriminative stimulus effects of morphine in squirrel monkeys: stimulants, opioids, and stimulant-opioid combinations. AB - Morphine and other mu opioids mimic and/or modulate the discriminative stimulus (DS) effects of cocaine, possibly reflecting mutual stimulation of mesolimbic dopamine activity. Less is known about the capacity of cocaine and related stimulants to modulate the DS effects of morphine. The present study investigated the effects of cocaine, amphetamine, and reference drugs, administered alone and with morphine, in squirrel monkeys trained to discriminate morphine from vehicle. Additional studies determined the ability of opioid and dopamine receptor antagonists to attenuate the DS effects of morphine and the morphine-like effects of other drugs. The DS effects of morphine were mimicked by the mu-opioid agonist fentanyl but not the delta-opioid agonists SNC 80 and BW 373U86 or the kappa opioid agonist U50,488H, and were antagonized by the opioid antagonist naltrexone but not the dopamine antagonist flupenthixol. In three of five monkeys, the DS effects of morphine also were mimicked by cocaine, amphetamine, and the dopamine transport inhibitor GBR 12909 but not the norepinephrine transport inhibitor talsupram or the serotonin transport inhibitor fluoxetine, and were antagonized by flupenthixol but not naltrexone. In this subgroup, pretreatment with cocaine or amphetamine enhanced the DS effects of morphine, whereas in the other two monkeys pretreatment with either stimulant attenuated the DS effects of morphine. The results demonstrated individual differences in morphine-like DS effects of stimulants that are mirrored by individual differences in their interactions with morphine. Furthermore, different mechanisms appear to mediate the DS effects of morphine and the morphine-like DS effects of cocaine and amphetamine. PMID- 10454483 TI - Glutathione conjugate interactions with DNA-dependent protein kinase. AB - A photoactivatable glutathione-drug conjugate (35)S-labeled-azidophenacyl glutathione (APA-SG) was synthesized and used to identify protein(s) involved in recognition and/or transport of glutathione conjugates of electrophilic drug species. A approximately 460-kDa protein was found to be highly labeled by (35)S labeled APA-SG in an Adriamycin-resistant HL-60 (HL-60/ADR) cell line and identified as the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) by amino acid sequence analysis, Western blot, and immunoprecipitation with specific antibodies. Binding specificity was confirmed by competition isotope dilution assays with purified proteins. A 15- to 20-fold increase in DNA-PKcs expression in the HL-60/ADR cell line was accompanied by an equivalent increase in (35)S labeled APA-SG binding. APA-SG, along with other glutathione conjugates and analogs inhibited the DNA-PK-mediated phosphorylation of an in vitro peptide substrate in a concentration-dependent manner. Using different antibodies to immunoprecipitate the individual components of the DNA-PK complex (DNA-PKcs, Ku70, and Ku80), it was shown that APA-SG caused a destabilization of the trimeric holoenzyme complex by dissociating the catalytic subunit from the Ku heterodimer. These data suggest that the kinase-mediated signaling is inhibited when glutathione conjugates bind to DNA-PKcs and may also indicate a possible strategy for design of novel DNA-PK inhibitors. PMID- 10454484 TI - Carrier-mediated hepatic uptake of peptidic endothelin antagonists in rats. AB - The endothelin antagonist BQ-123, an anionic cyclopentapeptide, is taken up by rat hepatocytes through active transport systems. Here, we have examined the hepatocellular uptake mechanism for several BQ-123 derivatives with anionic charges using isolated rat hepatocytes. BQ-485, a linear peptide, BQ-518, a cyclic peptide, and compound A, a cyclic peptide with a cationic moiety, were taken up by hepatocytes in a concentration-dependent manner. The uptake of BQ-485 was most efficient, whereas compound A showed comparable uptake with BQ-123. The uptake of these peptides was Na(+)- and energy-dependent, suggesting that active transport mechanisms are involved in their uptake into hepatocytes. BQ-485, BQ 518, and compound A can almost completely inhibit both the Na(+)-dependent and independent uptake of [(3)H]BQ-123, with inhibition constants (K(i)) that are comparable to the Michaelis-Menten constants (K(m)) for their Na(+)-dependent and -independent uptake, respectively. Inhibition by BQ-485 was competitive, and the uptake of BQ-485 can be inhibited by BQ-123, with K(i) values that are comparable with the K(m) values for BQ-123 uptake. The uptake of BQ-123 by COS-7 cells transfected with either Na(+)-dependent taurocholate-cotransporting polypeptide (Ntcp) or Na(+)-independent basolateral organic anion-transporting polypeptide (oatp1) was minimal. Thus, these three peptides share the transporters that also recognize BQ-123 but appear to differ from Ntcp and oatp1. PMID- 10454485 TI - Diltiazem inhibition of cytochrome P-450 3A activity is due to metabolite intermediate complex formation. AB - Diltiazem (DTZ) N-demethylation occurs by cytochrome P-450 (CYP) 3A based on the following observations: 1) a single enzyme Michaelis-Menten model of metabolite formation, 2) high correlations of DTZ N-demethylation activity to other CYP3A activities, 3) inhibition of DTZ N-demethylation activity by triacetyloleandomycin, and 4) DTZ N-demethylation activity by expressed CYP3A enzymes only. The mean K(m)s for DTZ N-demethylation in human liver microsomes and expressed CYP3A4(+b(5)) were 53 and 16 microM, respectively. A 30-min preincubation of DTZ in expressed CYPs inhibited CYP3A4(+b(5)) by 100%, of which 55% was due to formation of a metabolite intermediate complex (MIC), which is an inactive form of CYP. MIC was observed in human liver microsomes and cDNA expressed CYP3A only. In experiments to assess simultaneous MIC formation and loss of CYP3A activity, DTZ caused greater than 80% inhibition of midazolam hydroxylation after a 60-min preincubation in human liver microsomes. The rate constants for MIC formation and loss of midazolam hydroxylation activity were equivalent for the line of best fit for both data sets, which illustrates that MIC formation causes the inhibition of CYP3A activity. The mechanistic inhibition was characterized in expressed CYP3A4(+b(5)), which exhibited a concentration dependent formation of MIC by DTZ (1-100 microM) with an estimated k(inact) of 0.17 min(-1) and K(I) of 2.2 microM. The partition ratio for expressed CYP3A4(+b(5)) was substrate concentration dependent and varied from 13 to 86. This study showed that DTZ inhibition of CYP3A substrate metabolism occurs primarily by MIC formation. PMID- 10454486 TI - Calcium channels involved in K+- and veratridine-induced increase of cytosolic calcium concentration in human cerebral cortical synaptosomes. AB - Human cerebral cortical synaptosomes were used to study voltage-dependent Ca(2+) channels mediating calcium influx in human axon terminals. Synaptosomes were depolarized by elevation of the extracellular K(+) concentration by 30 mM or by the addition of veratridine (10 microM). Increase in cytosolic concentration of calcium [Ca(2+)](i) induced by either stimulus was abolished in the absence of extracellular Ca(2+) ions. omega-Agatoxin IVA inhibited the K(+)-induced [Ca(2+)](i) increase concentration-dependently (IC(50): 113 nM). omega-Conotoxin GVIA (0.1 microM) inhibited K(+)-induced [Ca(2+)](i) increase by 20%. omega Conotoxin MVIIC (0.2 microM) caused an inhibition by 85%. Nifedipine (1 microM) had no effect on K(+)-induced [Ca(2+)](i) increase. Veratridine-induced increase in [Ca(2+)](i) was inhibited by omega-conotoxin GVIA (0.1 microM) and omega Agatoxin IVA (0.2 microM; by about 25 and 45%, respectively). Nifedipine inhibited the veratridine-evoked [Ca(2+)](i) increase concentration-dependently (IC(50): 4.9 nM); Bay K 8644 (3 microM) shifted the nifedipine concentration response curve to the right. Mibefradil (10 microM) abolished the increase in [Ca(2+)](i) evoked by K(+) and reduced the increase evoked by veratridine by almost 90%. KB-R7943 (3 microM) an inhibitor of the Na(+)/Ca(2+) exchanger NCX1, decreased the increase in [Ca(2+)](i) evoked by veratridine by approximately 20%. It is concluded that the increase in [Ca(2+)](i) after K(+) depolarization caused by Ca(2+) influx predominantly via P/Q-type Ca(2+) channels and after veratridine depolarization via N- and P/Q-type, but also by L-type Ca(2+) channels. The toxin and nifedipine-resistant fraction of the veratridine response may result both from influx via R-type Ca(2+) channels and by Ca(2+) inward transport via Na(+)/Ca(2+) exchanger. PMID- 10454487 TI - Opioid antinociception in ovariectomized monkeys: comparison with antinociception in males and effects of estradiol replacement. AB - Baseline nociception and opioid antinociception were compared in male and ovariectomized female rhesus monkeys. Females were studied without estradiol replacement or during treatment with estradiol benzoate at doses (0.002 and 0.01 mg/kg/day) designed to mimic 17beta-estradiol blood levels observed during different phases of the menstrual cycle and during pregnancy. Baseline sensitivity to thermal stimuli (42-54 degrees C) was similar in male and ovariectomized female monkeys. The antinociceptive effects of the mu-opioid agonists fentanyl, morphine, butorphanol, and nalbuphine were examined at 50 and 54 degrees C. There were no sex-related differences in the antinociceptive effects of the high-efficacy mu agonist fentanyl; however, the lower-efficacy mu agonists morphine, butorphanol, and nalbuphine produced greater antinociceptive effects in males than in untreated ovariectomized females. Because butorphanol and nalbuphine have low selectivity for mu versus kappa receptors and may produce kappa-agonist effects under some conditions, the high-efficacy, kappa-selective agonist U50,488 was also studied. U50,488 also produced greater antinociceptive effects in males. Treatment with estradiol benzoate tended to enhance opioid antinociception in the ovariectomized females; however, this effect was significant only for butorphanol and U50,488 during treatment with the highest dose of estradiol benzoate. These findings suggest that opioid agonists usually produce greater antinociception in male monkeys than in females, and the magnitude of these sex-related differences may be inversely related to efficacy at mu receptors or selectivity for mu versus kappa receptors. Estradiol appears to have little effect on mu-agonist antinociception in primates but may enhance the antinociceptive effects of kappa agonists. PMID- 10454488 TI - Structure-function analysis of human cytochrome P-450 2B6 using a novel substrate, site-directed mutagenesis, and molecular modeling. AB - The structural basis for functional differences between human cytochrome P-450 2B6 and rat 2B1 was investigated. An amino acid sequence alignment predicted the location of 2B6 substrate recognition site (SRS) residues. Ten residues within these SRSs unique to 2B6 compared with 2B1, 2B4, and 2B11 were chosen for mutagenesis. Two additional sites that differ between 2B6 and 2B1 and are known to have a role in 2B1 substrate specificity were also mutated. The 2B6 mutants were expressed in Spodoptera frugiperda cells and characterized using the 2B6 specific substrate RP 73401 [3-cyclopentyloxy-N-(3,5-dichloro-4-pyridyl)-4 methoxybenzamide], the 2B1-selective substrate androstenedione, and the common substrate 7-ethoxy-4-trifluoromethylcoumarin. Mutants F107I and L363V exhibited decreased RP 73401 hydroxylation but retained most of the wild-type level of 2B6 7-ethoxy-4-trifluoromethylcoumarin O-deethylase activity. In addition, SRS exchanges were studied in which the amino acid sequence of 2B6 SRSs was converted to the sequence of 2B1. Each of these constructs, having two to seven substitutions, expressed at levels similar to 2B6 but did not acquire significant androstenedione hydroxylase activity. Docking of RP 73401 into the active site of a 2B6 homology model suggested a direct interaction with residue L363 but not with F107. Findings from this study suggest that 1) residues F107 and L363 are necessary for 2B6 RP 73401 hydroxylase activity, 2) 2B6 is able to tolerate multiple SRS substitutions without compromising protein expression levels or protein stability, and 3) conferring androstenedione hydroxylase function to cytochrome P-450 2B6 is more complex than altering a single SRS. PMID- 10454489 TI - Preclinical evaluation of newly approved and potential antiepileptic drugs against cocaine-induced seizures. AB - Seizures and status epilepticus are among the neurological complications of cocaine overdose in humans. The aim of the present study was to evaluate the protective effectiveness and therapeutic index (separation between anticonvulsive and side effect profiles) of 14 newly approved and potential antiepileptic drugs using a murine model of acute cocaine toxicity and the inverted-screen test for behavioral side effect testing. Cocaine (75 mg/kg i.p.) produces clonic seizures (approximately 90% of mice), and conventional antiepileptic drugs have been reported to be either ineffective or only effective at doses producing significant sedative/ataxic effects. Clobazam, flunarizine, lamotrigine, topiramate, and zonisamide were ineffective against seizures up to doses producing significant motor impairment. In contrast, felbamate, gabapentin, loreclezole, losigamone, progabide, remacemide, stiripentol, tiagabine, and vigabatrin produced dose-dependent protection against cocaine-induced convulsions with varied separations between their anticonvulsant and side effect profiles: the protective index values (toxic TD(50)/anticonvulsive ED(50)) ranged from 1.26 (felbamate) to 7.67 (loreclezole), and gabapentin had the highest (protective index >152). Thus, several drugs were identified with greater protective efficacy and reduced motor impairment compared with classic antiepileptic drugs. Based on the proposed mechanism of action of these new anticonvulsants, it is noteworthy that 1) drugs that enhance gamma-aminobutyric acid-mediated neuronal inhibition in a manner distinct from barbiturates and benzodiazepines offer the best protective/behavioral side effect profiles, and 2) functional antagonists of Na(+) and Ca(2+) channels are generally ineffective. Overall, this study provides the first description of the effectiveness of new antiepileptic drugs against experimentally induced cocaine seizures and points to several drugs that deserve clinical scrutiny for this indication. PMID- 10454490 TI - Discriminative stimulus effects of the nonpeptidic delta-opioid agonist SNC80 in rhesus monkeys. AB - Five rhesus monkeys were trained to discriminate the nonpeptidic, delta-opioid agonist SNC80 (0.32 mg/kg i.m.) from saline by using a food-reinforced drug discrimination procedure. Cumulative doses of SNC80 produced a dose-dependent increase in SNC80-appropriate responding and a dose-dependent decrease in response rate. In time-course studies, peak effects of the training dose of SNC80 were observed after 15 min, and these effects diminished over 240 min. In substitution studies, other piperazinyl benzamide delta agonists (SNC86, SNC162, and SNC243A) substituted for SNC80 with relative potencies similar those of SNC80. However, SNC67, the (-)-enantiomer of SNC80, did not occasion SNC80 appropriate responding up to a dose (32.0 mg/kg) that produced convulsions in one monkey. The mu agonists morphine and fentanyl and the kappa agonists U-50,488 and enadoline failed to substitute for SNC80 up to doses that eliminated responding. Two nonopioids (the N-methyl-D-aspartate antagonist ketamine and the monoamine reuptake inhibitor cocaine) also produced primarily saline-appropriate responding. Both the discriminative stimulus and rate-decreasing effects of SNC80 were antagonized by the delta-selective antagonist naltrindole (0.01-1.0 mg/kg) but not by doses of the opioid antagonist quadazocine (0.1-1.0 mg/kg) that block the effects of mu and kappa agonists. These data suggest that the discriminative stimulus effects of SNC80 are mediated by delta-opioid receptors and that the discriminative stimulus effects of delta opioids in primates can be differentiated from the effects of other opioid and nonopioid compounds. PMID- 10454491 TI - Inhibitors of calmodulin-dependent protein kinase are nonspecific blockers of voltage-dependent K+ channels in vascular myocytes. AB - The present study was undertaken to investigate the effects of specific inhibitors of calmodulin-dependent protein kinase II (CamKII) on macroscopic voltage-dependent K(+) current (K(V)) recorded from rabbit portal vein smooth muscle cells. Inhibition of L-type Ca(2+) current facilitation by 1 microM KN-62, a blocker of CamKII, was first demonstrated and provided evidence for functional CamKII activity in this preparation. KN-93, another specific and more potent inhibitor of CamKII in the rat brain, suppressed K(V) and enhanced the rate of inactivation in a dose-dependent manner, in cells dialyzed with both low (0.1 mM) and high (10 mM) EGTA pipette solution. Prolonged dialysis with 10 microM of a synthetic peptide inhibitor of CamKII (fragment 281-301) had little effect on K(V) and did not prevent the inhibitory action of KN-93 on the current. The estimated IC(50) for inhibiting peak and late currents during 250-ms steps to +60 mV (holding potential = -60 mV) were 2.9 and 0.27 microM, respectively. KN-93 also induced slight shifts of the steady-state activation (-7 mV) and inactivation (-6 mV) curves. KN-62, and KN-92, an inactive analog of KN-93, produced effects similar to those of KN-93. In current clamp experiments, 5 microM KN-93 depolarized the myocytes from a control resting membrane potential of -42.3 +/- 2.8 mV to -28.5 +/- 1.4 mV, an effect that was partially reversible after washout (-34.4 +/- 1.3 mV, n = 6). In conclusion, blockers of CamKII produce nonspecific inhibitory effects on K(V) that warrant cautious use of these compounds in physiological experiments designed to assess the role of CamKII. PMID- 10454492 TI - Contribution of nitric oxide to the presynaptic inhibition by endothelin ETB receptor of the canine stellate ganglionic transmission. AB - We previously reported that endothelin (ET) 3 inhibited presynaptically the dog stellate ganglionic transmission. Here, we report the investigation of the possible involvement of nitric oxide pathway in the endothelin-induced inhibition of the ganglionic transmission. The amount of acetylcholine released by preganglionic stimulation for 10 min was concentration-dependently inhibited after exposure to ET-3 (10(-9)-10(-6) M) or IRL-1620, endothelin ET(B) receptor agonist (10(-8)-10(-5) M). The inhibition was antagonized by pretreatment with a nonselective endothelin receptors antagonist (bosentan) and an ET(B) receptor antagonist (BQ-788) or a neuronal nitric oxide synthase inhibitor, 3-bromo-7 nitroindazole, but was not inhibited by a selective ET(A) receptor antagonist, BQ 123. The reduction induced by ET-3 was also antagonized by treatment with a selective inhibitor of soluble guanylyl cyclase, 1H-[1,2, 4]oxadiazolo[4,3 a]quinoxalin-1-one. In addition, similar reductions were also mimicked by exposure to cGMP analog, 8-bromoguanosine-3, 5-cyclic monophosphate and nitric oxide donor, S-nitroso-N-acetylpenicillamine. Exposure to ET-3 or IRL-1620 for a 30-min period increased the levels of total nitric oxide (NO), nitrite plus nitrate NO(x) concentration in the incubation medium, with the increase in NO(x) also being antagonized by BQ-788 at the same concentration. The ET-3-induced increase in NO(x) was antagonized by treatment with the same concentration of 3 bromo-7-nitroindazole or a selective inhibitor of receptor-mediated Ca(2+) entry, 1-[b-[3-(4-methoxyphenyl) propoxy]-4-methoxyphenethyl]-1H-imidazole (10(-5) M), and with a calmodulin antagonist, N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide. These results indicate that ET(B) receptor activation inhibits the sympathetic ganglionic transmission via reducing acetylcholine release from presynaptic nerve terminals, although this inhibition also seems to involve the ET(B) receptor-operated Ca(2+)-calmodulin-dependent activation of endogenous nitric oxide production. PMID- 10454493 TI - Age- and growth hormone-induced alterations in renal sulfate transport. AB - The effects of growth hormone (GH) treatment on renal sodium sulfate cotransport (NaSi-1) were studied in adult (9-10 months) and old (22-23 months) male Fischer 344 rats. All animals received twice-daily s.c. injections of recombinant human GH (hGH; 4 mg/kg) for up to 6 days. Animals were sacrificed by exsanguination on days 0, 1, 2, 3, 4, 5, and 6. Kidneys were removed, and kidney cortex was trimmed immediately and used for RNA and membrane preparations. Plasma hGH concentrations were significantly lower in old rats during the hGH treatment (P <.05). Insulin like growth factor-I (IGF-I) levels were significantly increased and remained stable after day 2 of hGH treatment in both age groups (P <.05). There was no significant difference in plasma IGF-I levels between age groups. Plasma IGF-I binding protein 3 (IGFBP-3) concentrations were significantly higher in 9- to 10 month-old rats compared with that in 22- to 23-month-old animals (P <.001). There were no significant differences in plasma IGFBP-3 concentrations between days of hGH treatment. The NaSi-1 mRNA levels were significantly lower in 22- to 23-month old rats compared with that in 9- to 10-month-old animals (P <.001). The NaSi-1 mRNA levels were significantly increased on days 2 and 3 of hGH treatment (P <.05) and then gradually decreased to the control value. The NaSi-1 protein levels in old animals (22-23 months) were also significantly lower than that of 9 to 10-month-old animals and were significantly increased from day 2 of hGH treatment, reaching a maximum level on day 3 or 4 and then returning to the baseline level in both age groups. From these results, it was concluded that 1) NaSi-1 mRNA and protein levels are lower in old animals and increase in both adult and aged rats after hGH treatment, 2) plasma IGF-I levels are similar in adult and aged rats and increase after hGH treatment, and 3) plasma IGFBP-3 levels are lower in old rats and remain unchanged after hGH treatment. PMID- 10454494 TI - Effects of ifenprodil and baclofen on exercise-induced increase of myocardial oxygen demand in normotensive rats. AB - Central glutamatergic relays are known to be present in the central sympathetic pathways. Ifenprodil (an N-methyl-D-aspartate antagonist) and baclofen (a gamma aminobutyric acid(B) agonist) are both modulators of these synapses; we previously reported their ability to reduce the cardiovascular responses induced by a central hypothalamic stimulation in rabbits. In this work, we investigated the actions of chronic treatments with these two drugs on the increase of myocardial oxygen demand induced by exercise in normotensive rats. Moreover, their effects on the baroreceptor heart rate reflex were observed. Male normotensive WKY rats were treated with placebo (two groups), baclofen, or ifenprodil for 14 days. They were then submitted to a progressively increased exercise test on a treadmill. In another three groups of animals, the same treatment was applied but, at the end, a baroreflex study was performed by the injection of phenylephrine (vagal component of the reflex) and of sodium nitroprusside (sympathetic component). Ifenprodil and baclofen reduced by nearly 50% the level of the increase of the rate x pressure product during exercise as compared with control rats. This effect appeared to be mainly due to a reduction of the hypertensive response. In the same conditions, neither baclofen nor ifenprodil significantly altered the baroreceptor heart rate reflex. The fact that these two drugs are capable of reducing the myocardial oxygen demand encourages us to test them in a model of myocardial ischemia associated with sympathetic hyperactivity. PMID- 10454495 TI - 5-Hydroxytryptamine(1A) receptor activation enhances norepinephrine release from nerves in the rabbit saphenous vein. AB - Although serotonergic receptor agonists are known to modulate release of central serotonin, less is known about the ability of serotonin to alter neurotransmission in peripheral adrenergic nerves. The present study used field stimulation (40V, 0.7 ms duration, 1-16 Hz) to contract the rabbit saphenous vein, an effect that was abolished in the presence of tetrodotoxin and prazosin (10(-6) M), consistent with stimulation of neuronal norepinephrine release. Furthermore, the field-stimulated contraction was not altered by the 5 hydroxytryptamine (5-HT)(1B/1D) receptor antagonist GR127935 (10(-6) M), but was markedly inhibited by the 5-HT(1A) receptor antagonist WAY 100635 (10(-6) M). GR127935 (10(-8) M) inhibited contraction to sumatriptan, documenting that the concentration used was sufficient to block 5-HT(1B/1D-like) vascular receptors in this tissue. Likewise, WAY 100635 (10(-6) M) inhibited contraction to the 5 HT(1A) receptor agonists (+/-)-8-hydroxydipropylaminotetralin hydrobromide (8-OH DPAT) and LY238729, without altering contraction to norepinephrine or sumatriptan. Furthermore, both 8-OH-DPAT and LY228729 enhanced the contractile response to field stimulation (1. 0-8.0 Hz) and activated norepinephrine release in the absence of field stimulation. Contractile responses of the rabbit saphenous vein to both 5-HT(1A) receptor agonists were markedly inhibited by prazosin and dextrally shifted by WAY 100635, supporting the idea that the 5 HT(1A) receptor agonists were activating presynaptic 5-HT(1A) receptors to enhance norepinephrine release even in the absence of field stimulation. Thus, in the rabbit saphenous vein, 5-HT(1A) but not 5-HT(1B/1D) receptors enhanced neurotransmitter release from adrenergic nerves. These observations suggested that serotonergic nerves or other cell types in the saphenous vein are activated by field stimulation to release serotonin, which in turn activates presynaptic 5 HT(1A) receptors on adrenergic neurons to effect norepinephrine release. To support this hypothesis, serotonin levels were measured in the saphenous vein and were increased after pargyline pretreatment (30 mg/kg s.c.), decreased after dl-p chlorophenylalanine methyl ester pretreatment (300 mg/kg s.c.), and unaltered after pretreatment with 6-hydroxydopamine hydrobromide (100 mg/kg s.c.). Thus, we provide strong evidence for the 1) presence of serotonin and its direct synthesis independent of adrenergic nerves and 2) a novel excitatory effect of presynaptic 5-HT(1A) receptor activation on adrenergic nerves in a peripheral blood vessel. PMID- 10454496 TI - Comparative pharmacology of the nonpeptide neuromedin B receptor antagonist PD 168368. AB - The mammalian peptide neuromedin B (NMB) and its receptor are expressed in a variety of tissues; however, little is definitively established about its physiological actions because of the lack of potent, specific antagonists. Recently, the peptoid PD 168368 was found to be a potent human NMB receptor antagonist. Because it had been shown previously that either synthetic analogs of bombesin (Bn) or other receptor peptoid or receptor antagonists function as an antagonist or agonist depends on animal species and receptor subtype studied, we investigated the pharmacological properties of PD 168368 compared with all currently known Bn receptor subtypes (NMB receptor, gastrin-releasing peptide receptor, Bn receptor subtype 3, and Bn receptor subtype 4) from human, mouse, rat, and frog. In binding studies, PD 168368 had similar high affinities (K(i) = 15-45 nM) for NMB receptors from each species examined, 30- to 60-fold lower affinity for gastrin-releasing peptide receptors, and >300-fold lower affinity for Bn receptor subtype 3 or 4. It inhibited NMB binding in a competitive manner. PD 168368 alone did not stimulate increases in either intracellular calcium concentration or [(3)H]inositol phosphates in any of the cells studied but inhibited NMB-induced responses with equivalent potencies in cells containing NMB receptors. PD 168368 was only minimally soluble in water. When hydroxypropyl-beta cyclodextrin rather than dimethyl sulfoxide was used as the vehicle, both the affinity and the antagonist potency of PD 168368 were significantly greater. The results demonstrate that PD 168368 is a potent, competitive, and selective antagonist at NMB receptors, with a similar pharmacology across animal species. PD 168368 should prove useful for delineating the biological role of NMB and selectively blocking NMB signaling in bioassays and as a lead for the development of more selective nonpeptide antagonists for the NMB receptor. PMID- 10454497 TI - A conditionally immortalized epithelial cell line for studies of intestinal drug transport. AB - A new cell culture model that better mimics the permeability of the human small intestine was developed for studies of passive drug transport. The intestinal epithelial cell line, 2/4/A1, conditionally immortalized with a temperature sensitive mutant of the growth-promoting oncogene simian virus 40 (SV40) large T, was grown on permeable supports. The cells grew at 33 degrees C, where the oncogene is fully active, but stopped growing and entered a differentiation program at 39 degrees C, where the oncogene is inactive. Significant cell death was observed at 39 degrees C and, therefore, growth conditions under which 2/4/A1 cells survive during the differentiation process were developed. Cells grown on extracellular matrices which contained laminin at an intermediate temperature of 37 degrees C formed viable differentiated monolayers with tight junctions, an increased expression of brush border enzymes, and a paracellular permeability that was comparable to that of the human small intestine. The permeability of 17 structurally diverse drugs gave a sigmoidal relationship with the absorbed fraction of the drugs after oral administration to humans. The relationship was compared with those obtained with the well established Caco-2 model and after in vivo perfusion of the human jejunum. The transport of drugs with low permeability in 2/4/A1 monolayers was comparable to that in the human jejunum, and up to 300 times faster than that in Caco-2 monolayers. The transport of drugs with high permeability was comparable in all models. These results indicate that 2/4/A1 monolayers are promising alternatives to Caco-2 monolayers for studies of passive drug transport. PMID- 10454498 TI - Apparent pA2 values of benzodiazepine antagonists and partial agonists in monkeys. AB - Drugs that bind to benzodiazepine recognition sites of gamma-aminobutyric acid type A receptor complexes may function as agonists in some behavioral assays and as antagonists in other behavioral assays. The present studies compared the effects of the benzodiazepines midazolam, flumazenil, bretazenil, Ro 41-7812, and Ro 42-8773 and the beta-carboline, beta-carboline-3-carboxylate-t-butyl ester (beta-CCt) under two different types of schedule-controlled responding in squirrel monkeys. One group of monkeys responded under a fixed-ratio schedule of stimulus-shock termination, and a second group of monkeys responded under a multiple fixed-ratio schedule of food presentation involving suppressed and nonsuppressed behavior. Under the schedule of stimulus-shock termination, midazolam produced dose-related decreases in response rate, and these effects were surmountably antagonized by flumazenil, bretazenil, Ro 41-7812, Ro 42-8773, and beta-CCt. Schild plot analysis of these data revealed the following mean pA(2) values: flumazenil, 7.18; bretazenil, 7.62; Ro 41-7812, 7. 06; Ro 42-8773, 6.95. Apparent pA(2) values were not calculated for beta-CCt because the CL of the slope of the Schild plot included positive values. Under the multiple schedule, midazolam, bretazenil, and Ro 42-8773 dose-dependently increased rates of suppressed responding, whereas flumazenil, Ro 41-7812, and beta-CCt had no significant rate-altering effects. Flumazenil antagonized the antisuppressant effects of midazolam and bretazenil; however, individual variability in these effects prohibited the determination of apparent pA(2) values. These results indicate that in vivo pA(2) values may be determined for benzodiazepine-site ligands. These results further demonstrate that some benzodiazepine-site ligands, e. g., bretazenil and Ro 42-8773, may function as both agonists and as competitive antagonists in vivo. PMID- 10454499 TI - Bimolecular glutathione conjugation kinetics of ethacrynic acid in rat liver: in vitro and perfusion studies. AB - The conjugation kinetics of glutathione (GSH) and ethacrynic acid (EA) were studied in rat liver perfusion studies, where efficient removal occurred (steady state extraction ratio E(ss), approximately 0.8-0.4 at concentrations ranging from 10-200 microM) despite the appreciable plasma protein binding. The declining E(ss) paralleled the saturation in GSH conjugate (EA-SG) formation; EA-SG primarily appeared in bile as the unchanged glutathionyl adduct (90%) and minimally as cleavage products. The GSH conjugation of EA in perfused liver was described by the constants K(m)(overall) of 67 microM and V(max)(overall) of 0.23 micromol/min/g liver. These differed from those observed for the bimolecular nonenzymatic (constant of 126 microM(-1) min(-1)) and enzymatic (K(m) for GSH and EA were 1.2 mM and 94 microM, respectively; V(max) of 533 nmol/min/mg liver cytosolic protein or 32 micromol/min/g liver) GSH conjugation of EA in vitro. But they were similar to those estimated for EA uptake in isolated rat hepatocytes by saturable (K(m)(uptake) = 57 microM, and V(max)(uptake) = 0.55 micromol/min/g liver) and nonsaturable (0.015 ml/min/mg) processes. At increasing EA concentrations (>25 microM), time-dependent changes were observed for E(ss) and EA-SG formation, which rapidly decreased with time after the attainment of steady state due to the rapid loss of cellular GSH. The composite data were described adequately by a physiological model that accounted for transport and the GSH dependent conjugation of EA. The results suggest that the rate-limiting process for hepatic EA GSH conjugation is cellular uptake, but cosubstrate availability controls the rate of metabolism when GSH becomes depleted. PMID- 10454500 TI - Development of a hepatocyte-specific prostaglandin E(1) polymeric prodrug and its potential for preventing carbon tetrachloride-induced fulminant hepatitis in mice. AB - A polymeric prodrug of prostaglandin E(1) (PGE(1)) was synthesized using galactosylated poly(L-glutamic acid hydrazide) (Gal-HZ-PLGA) as a biodegradable and targetable carrier to hepatocytes. Poly(L-glutamic acid hydrazide) was prepared by reacting poly(gamma-benzyl-L-glutamate) with hydrazine monohydrate, followed by reaction with 2-imino-2-methoxyethyl-1-thiogalactosides to obtain Gal HZ-PLGA after i.v. injection. (111)In-labeled galactosylated poly(L-glutamic acid hydrazide) ((111)In-Gal-HZ-PLGA) rapidly accumulated in the liver in a dose dependent manner, whereas (111)In-poly(L-glutamic acid hydrazide) did not, indicating the involvement of a galactose-specific mechanism in the uptake of (111)In-Gal-HZ-PLGA. Fractionation of liver cells revealed that (111)In-Gal-HZ PLGA was preferentially taken up by liver parenchymal cells. After being taken up by the liver, (111)In-Gal-HZ-PLGA was gradually degraded, and radioactive metabolites with low molecular weight were detected within 10 min after injection. Then, PGE(1) or [(3)H]PGE(1) was coupled to Gal-HZ-PLGA via a hydrazone bond under mild conditions to obtain PGE(1) conjugate. After i.v. injection, [(3)H]PGE(1) conjugate was rapidly taken up by the liver (more than 80% of the dose). After injection of the conjugate, (3)H radioactivity remained in the liver, representing about 70% of the dose, even at 24 h, whereas little radioactivity remained in the organ at 1 h after the injection of free [(3)H]PGE(1). Finally, its pharmacological activity was examined in mice with fulminant hepatitis induced by peritoneal injection of carbon tetrachloride. The i.v. injection of PGE(1) conjugate at a dose of 1 mg (0.074 mg PGE(1))/kg effectively inhibited the increase of plasma glutamic pyruvic transaminase activity, whereas twice this dose (0.15 mg/kg) of free PGE(1) had little effect. These results suggest that the PGE(1) conjugate is an excellent polymeric prodrug of PGE(1) for hepatitis therapy. PMID- 10454501 TI - Modulation of cytochrome P-450 gene expression in endotoxemic mice is tissue specific and peroxisome proliferator-activated receptor-alpha dependent. AB - Administration of the bacterial endotoxin lipopolysaccharide (LPS) causes induction of cytochrome P-450 (CYP) 4A mRNAs in rat liver and kidney. Because induction of the CYP4A subfamily by chemicals requires peroxisome proliferator activated receptor-alpha (PPARalpha), we determined whether CYP4A induction by LPS also requires PPARalpha by comparing the responses of PPARalpha-null (-/-) and wild-type (+/+) mice. Renal expression of CYP4A10, CYP4A14, and acyl-CoA oxidase was induced by LPS treatment in (+/+) mice, and these effects were absent in the (-/-) mice. In contrast, hepatic expression of CYP4A10 was down-regulated in the (+/+) animals, and no significant induction of acyl-CoA oxidase or CYP4A14 was detected in liver. Expression of the peroxisomal bifunctional enzyme was not significantly affected by LPS treatment. These results indicate that PPARalpha is activated in mouse kidney after LPS treatment and that this leads to modulation of some PPARalpha-regulated genes. However, the species and tissue specificity of these effects suggest that inflammatory pathways may modulate the induction via PPARalpha. Mice pair fed with LPS-treated mice showed no induction of renal CYP4A10 or CYP4A14, indicating that renal CYP4A induction during endotoxemia is not due to hypophagia. Down-regulation of CYP2A5, CYP2C29, and CYP3A11 by LPS was attenuated or blocked in the (-/-) mice, suggesting a role for PPARalpha in CYP down-regulation as well. Finally, we found that clofibrate caused an acute induction of two hepatic acute-phase mRNAs that was only partially dependent on PPARalpha. PMID- 10454502 TI - Effect of several 5-hydroxytryptamine(1A) receptor ligands on the micturition reflex in rats: comparison with WAY 100635. AB - Several novel N-arylpiperazine derivatives were synthesized and tested for their 1) affinity and functional activity on 5-hydroxytryptamine(1A) (5-HT(1A)) receptors in vitro; 2) activity in models predictive of antagonism at somatodendritic and postsynaptic 5-HT(1A) receptors; and 3) effects on the micturition reflex in anesthetized and conscious rats. These studies also included 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine hydrobromide (NAN 190), 8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4, 5]decane 7,9-dione dihydrochloride (BMY 7378), and N-[2-[4-(2-methoxyphenyl)-1 piperazinyl]ethyl-N-(2-pyridinyl)cyclohex anecarboxamide (WAY 100635). Almost all compounds were found to be potent and selective for the human recombinant 5 HT(1A) receptor, with K(i) values in the nanomolar range. [(35)S]GTPgammaS binding in HeLa cells expressing the recombinant human 5-HT(1A) receptor allowed classification of the compounds into neutral antagonists and partial agonists. Almost all neutral antagonists were active in blocking 8-hydroxy-2 dipropylaminotetralin (8-OH-DPAT)-induced forepaw treading in rats (postsynaptic model) and hypothermia in mice (somatodendritic model) with the same potency, whereas compounds showing partial agonistic activity were active in the postsynaptic model but were inactive, or poorly active, in the somatodendritic model. Neutral antagonists potently inhibited volume-induced bladder-voiding contractions in anesthetized rats. Contractions were completely blocked, and the disappearance of bladder contractions lasted 7 to 13 min after the highest doses tested. Furthermore, neutral antagonists increased bladder volume capacity in conscious rats during continuous transvesical cystometry, whereas micturition pressure was only slightly, and not dose-dependently, reduced. Partial agonists were inactive or poorly active, inducing a disappearance time of bladder contractions that did not exceed 6 min in anesthetized rats, and failing to increase bladder volume capacity in conscious rats. These findings indicate that only neutral 5-HT(1A) receptor antagonists are endowed with inhibitory effects on the bladder. PMID- 10454503 TI - Involvement of cyclic nucleotide- and calcium-regulated pathways in phenobarbital induced cytochrome P-450 3A expression in mouse primary hepatocytes. AB - Several of the hepatic microsomal cytochromes P-450 (CYP) including CYP3A are inducible by phenobarbital (PB). However, the intracellular pathways involved in the action of PB on CYP3A remain poorly known. With the aim to unravel some of the main aspects of PB signaling, we first devised a simple model of mouse cultured primary hepatocytes in which CYP3A mRNA and protein were strongly induced by PB in the absence of dexamethasone and were at maximum levels after a 48-h treatment with a 2-mM dose of PB. Under these culture conditions, we studied the effects of inhibitors and activators of different protein kinases or phosphatases on CYP3A mRNA and protein induction by PB. CYP3A-induced expression was inhibited by activators of cyclic AMP-dependent protein kinase (PKA) (dibutyryl-cyclic AMP and forskolin) whereas inhibition of PKA by PKA inhibitor enhanced induction. 8-br-cGMP produced effects similar to the activators of PKA, and so did the specific inhibitor of cGMP-dependent protein kinase, beta-phenyl 1, N(2)-etheno-8-bromoguanosine-3,5'-cyclic monophosphorothioate, Rp-isomer (Rp-8 Br-PET-cGMPS). Inhibition of Ca(2+)/calmodulin-dependent protein kinase by KN-62 or the intracellular Ca(2+) chelator BAPTA-AM produced an inhibition of CYP3A induction by PB. Specific inhibitors of protein kinase C, mitogen-activated protein kinase kinase, phosphatidylinositol-3-kinase, or serine/threonine phosphatase did not produce any effect. Taken together, our results suggest that CYP3A induction by PB is regulated positively by calmodulin-dependent protein kinase and cGMP-dependent protein kinase, and negatively by PKA in mouse hepatocytes in primary culture. PMID- 10454504 TI - AL-8810: a novel prostaglandin F2 alpha analog with selective antagonist effects at the prostaglandin F2 alpha (FP) receptor. AB - A novel analog of prostaglandin F(2alpha) [AL-8810; (5Z, 13E)-(9S, 11S,15R)-9,15 dihydroxy-11-fluoro-15-(2-indanyl)-16,17,18,19, 20-pentanor-5,13-prostadienoic acid] has been discovered with uniquely low efficacy (E(max)) at the endogenous prostaglandin F(2alpha) receptors (FP receptors) of A7r5 rat thoracic aorta smooth muscle cells and Swiss mouse 3T3 fibroblasts, as assayed by stimulation of phospholipase C activity. AL-8810 has weak agonist potency (EC(50)) of 261 +/- 44 nM (n = 3) and E(max) = 19% (relative to the full FP receptor agonist cloprostenol) in A7r5 cells and EC(50) of 186 +/- 63 nM (n = 3) and E(max) = 23% in 3T3 fibroblasts. AL-8810 exhibited properties of an apparent competitive antagonist, i.e., produced parallel dextral shifts of the agonist concentration response curves and no significant suppression of the maximal agonist-induced response, when the potent, selective FP receptor agonist fluprostenol was used. The inhibition parameters of AL-8810 were: pA(2) = 6.68 +/- 0.23 and 6.34 +/- 0.09 (n = 3-4) for A7r5 cells and 3T3 cells, respectively, with Schild slopes ranging from 0.80 to 0.92. AL-8810 concentration-dependently antagonized the response to 100 nM fluprostenol (K(i) = 426 +/- 63 nM; n = 5) in A7r5 cells. However, even at 10 microM concentration, AL-8810 did not significantly inhibit functional responses of TP, DP, EP(2), EP(4), receptor subtypes in various cell lines. AL-8810 also did not antagonize the phospholipase C-coupled V(1) vasopressin receptor in A7r5 cells. These results suggest that AL-8810 is a unique, selective antagonist at the FP receptor, a heretofore unavailable pharmacological tool that should be valuable for studying FP receptor-mediated functional responses in complex biological systems. PMID- 10454505 TI - Effects of lipid mediator antagonists on predominant mediator-controlled asthmatic reactions in passively sensitized guinea pigs. AB - The role of cysteinyl leukotrienes (cys-LTs) and thromboxane A(2) (TXA(2)) in guinea pig models of aspects of bronchial asthma was investigated. In a novel antigen (BSA)-induced asthmatic model using passively sensitized guinea pigs, pretreatment with varying doses of indomethacin controlled the ratio of followed lipid mediators, LTC(4)/D(4)/E(4) and TXB(2), in lungs of challenged guinea pigs. The predominant mediator in indomethacin-untreated asthma was TXA(2), and complete inhibition of cyclooxygenase by i.v. injection of 5-mg/kg indomethacin induced cys-LTs mainly mediated asthmatic response. Furthermore, a 1-mg/kg indomethacin dose induced an asthmatic state where both cys-LTs and TXA(2) equally participated. Either LTD(4) or TXA(2) receptor antagonists given alone inhibited the asthmatic response in conditions where the corresponding mediator plays a predominant role. The combination of LTD(4) and TXA(2) receptor antagonists exhibited significant effects irrespective of the condition used. Under conditions where both mediators equally participate, a combination of both receptor antagonists showed additive inhibition. YM158, a newly synthesized and orally active dual antagonist for LTD(4) and TXA(2) receptors, showed the same antiasthmatic effect as a combinated LTD(4) receptor antagonist and a TXA(2) receptor antagonist mixture. Therefore, broad-acting compounds such as YM158 are expected to have antiasthmatic efficacies in a broader class of asthmatic patients than single-acting drugs. PMID- 10454506 TI - Sexual dimorphism in phencyclidine in vitro metabolism and pharmacokinetics in rats. AB - Studies were conducted to determine the differences in phencyclidine (PCP) in vitro metabolism and pharmacokinetics in female and male Sprague-Dawley (SD) rats. Formation rates of five major PCP metabolites in liver microsomes were significantly higher (p <.05) in males compared with females in three different rat strains (SD, Fischer 344, and Dark Agouti). In addition, the formation rate for an irreversibly bound PCP metabolite in males was the second highest of the six metabolites measured in these studies. However, the liver microsomes from the females produced essentially no metabolite binding in any strain. To determine the in vivo consequences of these in vitro metabolism results, we determined PCP's pharmacokinetic profile in female SD rats after a pharmacologically active i.v. dose of PCP (1 mg/kg) and then compared these data with the pharmacokinetic profile in male SD rats. The value for PCP systemic (and nonrenal) clearance was more than 45% lower (p <.05) in female rats. In addition, the terminal elimination T(1/2) was significantly longer (p <.05) in the female rats (5.5 versus 3.4 h, respectively). Because the initial serum concentration, volume of distribution at steady state, and renal clearance were not significantly different between the sexes, the longer half-life was attributed directly to a decreased ability of females to metabolize the drug. Consequently, these pharmacokinetic data suggest pharmacological differences in PCP effects between female and male rats are due primarily to a decreased ability of female rats to metabolize the drug. PMID- 10454507 TI - In vivo formation and localization of 1,1-dichloroethylene epoxide in murine liver: identification of its glutathione conjugate 2-S-glutathionyl acetate. AB - The hepatotoxic effects induced by 1,1-dichloroethylene (DCE) are ascribed to cytochrome P-450 (CYP) 2E1-dependent formation of metabolites including 2,2 dichloroacetaldehyde and the DCE-epoxide. The DCE metabolites detected in incubations of liver microsomes are the acetal, the hydrate of 2,2 dichloroacetaldehyde, and the epoxide-derived GSH conjugates 2-S-glutathionyl acetyl glutathione ([B]) and 2-S-glutathionyl acetate ([C]). This study was undertaken to determine whether these DCE metabolites are also formed in vivo in murine liver. HPLC analysis of cytosol isolated from the livers of mice treated with [(14)C]DCE showed that [C] was the major conjugate formed, with lower levels of formation of [B]. The acetal was not detected in the cytosol. The formation of the epoxide-derived GSH conjugates was dose-dependent at 25 to 225 mg/kg DCE and occurred coincidentally with levels of covalent binding of DCE at the same doses. The acetal and conjugates [B] and [C] were also detected in bile collected from mice treated with DCE. Pretreatment of mice with buthionine sulfoximine decreased sulfhydryl levels and formation of conjugate [C], and increased DCE binding to liver proteins. In contrast, the levels of [C] and DCE binding were both reduced significantly in mice pretreated with the CYP2E1 inhibitor diallyl sulfone. Immunohistochemical studies indicated that protein adducts and conjugate [C] were localized in centrilobular hepatocytes and corresponded with the sites where CYP2E1 resided. Pretreatment with buthionine sulfoximine increased the amount of immunostaining. However, pretreatment with diallyl sulfone markedly decreased immunostaining for [C] in the hepatocytes. These results showed that 2,2 dichloroacetaldehyde and the epoxide are formed from DCE in vivo. PMID- 10454508 TI - Modulation of neurotransmitter release in the basal ganglia of the rat brain by dynorphin peptides. AB - Microinjection studies have found that although dynorphin peptides decrease dopamine release in the rat basal ganglia, the nonselective opiate antagonist naloxone produces the opposite effect. To investigate the contribution of the dynorphin pathways to a tonic modulation of dopamine release, a microdialysis study was undertaken, with probes implanted in the substantia nigra and the ipsilateral neostriatum. Perfusion of the substantia nigra with the nonselective antagonist naltrexone (NTX; 1-10 microM), the selective kappa-opoid receptor antagonist, nor-binaltorphimine (nor-BNI; 1-10 microM), and the selective mu opioid receptor antagonist, D-Pen-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2) (CTOP; 1-10 microM) produced an increase in dopamine release, both in substantia nigra and neostriatum. nor-BNI also produced an increase in dynorphin B release, and a similar effect was observed with the higher concentration of NTX (10 microM). At the higher concentration of NTX and CTOP, an increase in glutamate release was also observed. Perfusion of the neostriatum with NTX, nor-BNI, or CTOP increased striatal dopamine, and dynorphin B release and increased dynorphin B in the ipsilateral substantia nigra. NTX and CTOP, but not nor-BNI, increased striatal glutamate and aspartate release. The kappa-opioid agonist U-50,488H (10 microM) induced a decrease in dopamine levels, both in the substantia nigra and neostriatum, and a paradoxical increase in striatal aspartate levels. Finally, systemic administration of NTX (4 mg/kg s.c.) in awake animals significantly increased striatal dopamine levels. The results suggest that opioid peptides, either dynorphins acting on kappa-opioid receptors or enkephalins acting on mu opioid receptors, exert tonic inhibition on dopamine and dynorphin B release in both substantia nigra and neostriatum. PMID- 10454509 TI - Effects of sex and gonadectomy on cocaine metabolism in the rat. AB - The purpose of the current study is to determine whether sex differences in metabolism of cocaine (COC) exist that could contribute to the greater behavioral sensitivity of females to COC administration. To investigate this question, concentrations of COC and its two principle metabolites benzoylecgonine (BE) and ecgonine methyl ester (EME) were measured by gas chromatography/mass spectroscopy in brain and plasma collected from male and female rats that were sacrificed between 5 and 90 min after injection COC (15 mg/kg i.p.). COC concentrations did not differ in plasma or brain tissue of males and females, but sex-specific patterns of metabolite distribution were detected. BE was 2-fold higher in plasma and brain of males than females, whereas EME was much higher in brain and plasma of females. The influence of gonadal hormones on COC metabolite patterns were determined using gonadectomized and prepubertal rats. Castration of male or female rats did not alter brain or plasma COC, but did decrease BE concentrations. Seven-day-old pups injected with 15 mg/kg of COC had higher blood and brain COC than adults and relatively low levels of metabolites. No sex differences were found for COC, BE, or EME in brain or plasma of pups. These findings indicate that although gonadal steroids influence COC metabolism, these effects do not explain sex differences in COC-induced behaviors. PMID- 10454510 TI - Species differences in the transport activity for organic anions across the bile canalicular membrane. AB - Species differences in the transport activity mediated by canalicular multispecific organic anion transporter (cMOAT) were examined using temocaprilat, an angiotensin-converting enzyme inhibitor whose biliary excretion is mediated predominantly by cMOAT, and 2,4-dinitrophenyl-S-glutathione, a typical substrate for cMOAT, in a series of in vivo and in vitro experiments. Temocaprilat was infused to examine the biliary excretion rate at steady-state. The in vivo transport clearance values across the bile canalicular membrane, defined as the biliary excretion rate divided by the hepatic unbound concentrations, were 9.8, 39.2, 9.2, 1.1, and 0.8 ml/min/kg for mouse, rat, guinea pig, rabbit, and dog, respectively. The K(m) and V(max) values for ATP-dependent uptake of 2, 4 dinitrophenyl-S-glutathione into canalicular membrane vesicles were 15.0, 29.6, 16.1, 55.8, and 30.0 microM and 0.38, 1.90, 0.15, 0. 47, and 0.23 nmol/min/mg protein, yielding the in vitro transport clearance across the bile canalicular membrane (V(max)/K(m)) of 25.5, 64.2, 9.4, 8.4, and 7.7 for mouse, rat, guinea pig, rabbit, and dog, respectively. A close in vivo and in vitro correlation was observed among animal species for the transport clearance across the bile canalicular membrane. These results suggest that the uptake experiments with canalicular membrane vesicles can be used to quantitatively predict in vivo excretion across the bile canalicular membrane. PMID- 10454511 TI - In vivo inhibition of peptidylglycine-alpha-hydroxylating monooxygenase by 4 phenyl-3-butenoic acid. AB - Peptidylglycine-alpha-hydroxylating monooxygenase (PHM; EC 1.14.17. 3) catalyzes the first and rate-limiting reaction in the two-step process that alpha-amidates neural and endocrine peptides. The substrate analog 4-phenyl-3-butenoic acid (PBA) was shown in vitro to selectively inhibit PHM without affecting the activity of peptidyl-alpha-hydroxyglycine alpha-amidating lyase, the enzyme that mediates the second reaction in alpha-amidation. Inhibition of PHM activity by PBA lowered the V(max) of the enzyme without altering its K(m). Administration of PBA in vivo profoundly inhibited serum PHM activity in a dose- and time-related fashion. Maximal reductions to less than 5% of control levels were observed 3 h after a single administration (500 mg/kg). Inhibition of serum PHM activity by PBA was short-lived, being fully reversed by 24 h postinjection. PHM activity in cardiac atrium, hypothalamus, and anterior and neurointermediate lobes of the pituitary were also decreased by PBA treatment but to a lesser extent than with serum. Inhibition of PHM activity by PBA was not cumulative over time when assessed 24 h after the last of 10 daily injections (500 mg/kg). The role of protein synthesis in maintaining PHM activity in blood was demonstrated by treatment with cycloheximide, which reduced serum PHM activity and retarded the recovery of PHM activity after PBA administration. It is concluded that the metabolism and/or clearance of PBA is rapid and that de novo protein synthesis has an important role in mediating the rapid restoration of PHM activity after PBA administration. PMID- 10454512 TI - Identification of quantitative trait loci for haloperidol-induced catalepsy on mouse chromosome 14. AB - Previous studies have established that neuroleptic-induced catalepsy in mice is a highly heritable trait. The current study focuses on the detection of quantitative trait loci (QTL) for haloperidol-induced catalepsy in a BALB/cJ x LP/J F(2) intercross. One thousand thirty-seven F(2) animals were phenotyped and divided into four categories: very responsive (RR), responsive, nonresponsive, and very nonresponsive (NN). The RR and NN phenotypes comprised approximately 18% each of the total and differed in their haloperidol sensitivity by >10-fold. Sex differed significantly between the NN and RR groups (chi(2) = 14.0; p <.0002); females comprised 58% of the RR individuals but only 38% of the NN individuals. The difference between the extreme phenotypes in the number of piebald animals was highly significant (chi(2) = 30, p <. 00001). Eight percent of the RR individuals were piebald compared with 30% of the NN individuals. A genome wide scan confirmed the presence of a QTL (peak LOD = 6.4) on chromosome 14 near the piebald (Ednrb) and 5-hydroxytryptamine(2A) (Htr2a) loci. Although the parental BALB/cJ and LP/J strains differed significantly in striatal 5 hydroxytryptamine(2A) receptor binding, no marked differences were detected between the phenotypic extremes. A second QTL was detected on chromosome 14 (peak LOD = 6.9), which was located more proximally and included the Chat locus. No QTLs were detected on chromosomes 1 and 9, thus differentiating this cross from previous results obtained for a C57BL/6J x DBA/2J intercross. PMID- 10454513 TI - Cocaine-reinforced responding in rhesus monkeys: pharmacological attenuation of the hypothalamic-pituitary-adrenal axis response. AB - Intravenously self-administered cocaine produces a dose-dependent release of adrenocorticotropic hormone (ACTH) and cortisol in male rhesus monkeys. This study investigated whether the acute disruption of cortisol and/or ACTH release had any effect on ongoing cocaine-maintained responding. Four hypothalamic pituitary-adrenal (HPA) axis inhibitors were examined: etomidate and ketoconazole, both of which are cortisol synthesis inhibitors; astressin, a peptidic corticotropin-releasing factor (CRF) antagonist that binds CRF(1) receptors predominantly in the pituitary gland; and dexamethasone, a highly selective glucocorticoid receptor agonist whose long-lasting effects reduce or abolish the endogenous release of ACTH and cortisol. The reinforcing effects of a range of cocaine doses, with or without pretreatment with an HPA inhibitor, were evaluated using a fixed ratio 30 time-out 10-min schedule of reinforcement in six male monkeys. Blood was sampled before, during, and after self-administration sessions. Self-administration of cocaine increased plasma cortisol and ACTH. Pretreatment with etomidate and ketoconazole dose-dependently inhibited the cocaine-induced rise in cortisol and, at the highest doses, produced a compensatory increase in ACTH release. Astressin and dexamethasone attenuated or abolished cocaine-induced cortisol and ACTH release. Despite the efficacy exhibited by these pretreatments and the variety of mechanisms by which they inhibited the HPA axis, there was no evidence for any change in cocaine reinforced behavior (response rate or infusion number), an indication that acute changes in the ACTH or cortisol response to cocaine do not play a direct role in modulating cocaine-seeking behavior under these behavioral circumstances. PMID- 10454514 TI - Divalent cations modulate N-methyl-D-aspartate receptor function at the glycine site. AB - The modulation of the N-methyl-D-aspartate (NMDA) receptor (NMDAR) by divalent cations was examined using (+)-5-methyl-10, 11-dihydro-5H-dibenzo[a,d]cyclohepten 5,10-imine maleate ([(3)H]MK-801) binding as a functional indicator of NMDAR function. Ca(2+) and Mg(2+) produce a biphasic effect on the binding of [(3)H]MK 801 to the NMDAR channel in extensively washed adult rat brain membranes. Concentrations of Ca(2+) and Mg(2+) between 1 and 600 microM potentiate binding, but higher concentrations inhibit binding. The potentiating effect of Ca(2+) and Mg(2+) on [(3)H]MK-801 binding is due to an increase in the maximal number of binding sites (B(max)) with no effect on binding affinity (K(d)). Ca(2+)- and Mg(2+)induced potentiation is the result of an apparent increase in the affinity of the NMDAR for glycine. The ontogeny of NMDAR potentiation by Ca(2+) and Mg(2+) was also investigated. The number of [(3)H]MK-801 binding sites associated with divalent cation potentiation are present at low levels shortly after birth, and increase to peak level at 17 days of age before declining to adult levels. The potency of Ca(2+) and Mg(2+) to stimulate [(3)H]MK-801 binding did not change as a function of age. Lead (Pb(2+)) and zinc (Zn(2+)), potent inhibitors of the NMDAR, antagonize NMDAR potentiation by Ca(2+) and Mg(2+). These findings indicate that divalent cations differentially regulate NMDAR function by modulation of the glycine site. The NMDAR glycine site may be important in the regulation of glutamatergic neurotransmission by physiologically and toxicologically relevant cations. PMID- 10454515 TI - Hyperforin, a major antidepressant constituent of St. John's Wort, inhibits serotonin uptake by elevating free intracellular Na+1. AB - Extracts of Hypericum perforatum (St. John's Wort) are widely used for the treatment of depressive disorders and are unspecific inhibitors of the neuronal uptake of several neurotransmitters. Previous studies have shown that hyperforin represents the reuptake inhibiting constituent. To characterize the mechanism of serotonin reuptake inhibition, kinetic analyses in synaptosomes of mouse brain were performed. Michaelis-Menten kinetics revealed that hyperforin (2 microM) induces a decrease in V(max) by more than 50% while only slightly decreasing K(m), indicating mainly noncompetitive inhibition. By contrast, citalopram (1 nM) leads to an elevation of K(m) without changing V(max). Monensin, a Na(+)/H(+) exchanger, showed similar properties as hyperforin (decrease of V(max) without changing K(m)). Compared with classical antidepressants, such as selective serotonin reuptake inhibitors and tricyclic antidepressants, hyperforin is only a weak inhibitor of [(3)H]paroxetine binding relative to its effects on serotonin uptake. As monensin decreases serotonin uptake by increasing Na(+)/H(+) exchange, we compared the effects of hyperforin and monensin on the free intracellular sodium concentration ([Na(+)](i)) in platelets by measuring 1,3 benzenedicarboxylic acid, 4,4'-[1,4,10-trioxa-7, 13-diazacyclopentadecan-7,13 diylbis(5-methoxy-6, 2-benzofurandiyl)]bis-, tetraammonium salt (SBFI/AM) fluorescence. Both drugs elevated [Na(+)](i) over basal levels, with a maximal [Na(+)](i) of 69 +/- 16.1 mM (50 microM hyperforin) and 140 +/- 9.1 mM (10 microM monensin). Citalopram at concentrations relevant for [(3)H]serotonin uptake inhibition had no effect on [Na(+)](i). Although the mode of action of hyperforin seems to be associated with elevated [Na(+)](i) similar to those levels found with monensin, the molecular mechanism might be different, as even at high concentrations, hyperforin does not elevate free intracellular sodium concentration ([Na(+)](i)) up to the extracellular level, as monensin does. Hyperforin represents the first substance with a known preclinical antidepressant profile that inhibits serotonin uptake by elevating [Na(+)](i). PMID- 10454516 TI - Baclofen inhibits heroin self-administration behavior and mesolimbic dopamine release. AB - An emerging hypothesis to explain the mechanism of heroin-induced positive reinforcement states that opiates inhibit gamma-aminobutyric acid (GABA)-ergic interneurons within the mesocorticolimbic dopamine (DA) system to disinhibit DA neurons. In support of this hypothesis, we report that the development of heroin self-administration (SA) behavior in drug-naive rats and the maintenance of SA behavior in heroin-trained rats were both suppressed when the GABA(B) receptor agonist baclofen was coadministered with heroin. Microinjections of baclofen into the ventral tegmental area (VTA), but not the nucleus accumbens, decreased heroin reinforcement as indicated by a compensatory increase in SA behavior. Additionally, baclofen administered alone or along with heroin dose-dependently reduced heroin-induced DA release. This effect was blocked partially by intra-VTA infusion of the GABA(B) antagonist 2-hydroxysaclofen, suggesting an additional, perhaps GABA(A) receptor-mediated, disinhibitory effect. Taken together, these experiments, for the first time, demonstrate that heroin-reinforced SA behavior and nucleus accumbens DA release are mediated predominantly by GABA(B) receptors in the VTA and suggest that baclofen may be an effective agent in the treatment of opiate abuse. PMID- 10454518 TI - Evidence that the apoptotic actions of etoposide are independent of c Jun/activating protein-1-mediated transregulation. AB - We recently demonstrated that physiological induction of apoptosis by cytotoxic sphingolipid messengers proceeds via activating protein-1 (AP1)-dependent and AP1 independent mechanisms in U937 human monoblastic leukemia cells. Here we examine involvement of the stress-activated protein kinase (SAPK) cascade and AP1 in the initiation of apoptosis in U937 cells by podophyllotoxin-derived inhibitors of topoisomerase II. Induction of apoptotic cell death and DNA damage by treatment of U937 cells with etoposide (100 microM) was associated with phosphorylation and activation of the c-Jun NH(2)-terminal kinase (JNK1) SAPK enzymes p46 and p54 JNK2 and transient increases in expression of the transcription factor c-Jun, a primary JNK substrate. These responses were accompanied by a modest, but sustained, recruitment of the mitogen-activated protein kinases p42-extracellular signal receptor-activated kinase (ERK)1 and p44-extracellular signal receptor activated kinase 2. The capacity of etoposide to promote double-stranded DNA degradation and cell death was unaffected by manipulations that interfere with SAPK signaling outflow through c-Jun/AP1, including: 1) pharmacological inhibition of AP1 activity by diferuloylmethane and 2) molecular ablation of normal c-Jun function by the Jun dominant-negative mutant TAM-67. Cytotoxicity of the structurally related compound teniposide was similarly unaffected. In parallel trials, the lethal actions of ceramide (but not of sphingosine) were markedly diminished by pretreatment with diferuloylmethane or expression of TAM 67, confirming the effectiveness of these interventions in suppression of SAPK/AP1-dependent apoptosis. The involvement of AP1 in the proapoptotic actions of other inhibitors of topoisomerase II activity was also evaluated. Induction of cell death by the anthracyclines daunorubicin, daunorubicin, and idarubicin was found to be insensitive to pretreatment with diferuloylmethane or expression of TAM-67. Collectively, the present data indicate that induction of apoptosis by etoposide and related inhibitors of topoisomerase II is mediated through a cell death pathway that does not require SAPK-dependent recruitment of AP1. These findings additionally suggest that activation of the SAPK represents a consequence, rather than an underlying cause, of etoposide-induced apoptosis in myeloid leukemia cells. PMID- 10454517 TI - Stimulatory effects of silibinin and silicristin from the milk thistle Silybum marianum on kidney cells. AB - The biochemical influence of flavonolignans from the milk thistle Silybum marianum has been tested on kidney cells of African green monkeys. Two nonmalignant cell lines were selected, with the focus of the work on the fibroblast-like Vero line. Proliferation rate, biosynthesis of protein and DNA, and the activity of the enzyme lactate dehydrogenase (as a measure of the cellular metabolic activity) were chosen as parameters for the effect of the flavonolignans. Silibinin and silicristin show remarkable stimulatory effects on these parameters, mainly in Vero cells; however, isosilibinin and silidianin proved to be inactive. In vitro experiments with kidney cells damaged by paracetamol, cisplatin, and vincristin demonstrated that administration of silibinin before or after the chemical-induced injury can lessen or avoid the nephrotoxic effects. The results warrant in vivo evaluations of the flavonolignan derivatives. PMID- 10454519 TI - Characterization of MK-801-induced behavior as a putative rat model of psychosis. AB - The objective of this study was to characterize the behavior induced by the N methyl-D-aspartate receptor antagonist MK-801 (dizocilpine maleate) in rats as a model of psychosis. The temporal profile, dose dependence, age, and sex differences of the behavior are described. A gas chromatographic method for the analysis of MK-801 in plasma and brain was developed. Female rats showed 4 to 10 times more MK-801-induced behavior and displayed around 25 times higher serum and brain concentrations of MK-801 than male rats. Twenty-one neuroactive compounds, including a number of excitatory amino acid-active substances, were tested for the effect on MK-801-induced behavior. Neuroleptics blocked MK-801-induced behavior in a dose-dependent manner that correlated to their antipsychotic potency in humans. Adenosine receptor agonists and an N-methyl-D-aspartate receptor-associated glycine site antagonist showed putative antipsychotic effects. In conclusion, MK-801-induced behavior represents a rat excitatory amino acid hypofunction model of psychosis that appears to be of clinical relevance and may be of value in the search for new antipsychotic agents. PMID- 10454520 TI - Biochemical and functional profile of a newly developed potent and isozyme selective arginase inhibitor. AB - An increase in arginase activity has been associated with the pathophysiology of a number of conditions, including an impairment in nonadrenergic and noncholinergic (NANC) nerve-mediated relaxation of the gastrointestinal smooth muscle. An arginase inhibitor may rectify this condition. We compared the effects of a newly designed arginase inhibitor, 2(S)-amino-6-boronohexanoic acid (ABH), with the currently available N(omega)-hydroxy-L-arginine (L-HO-Arg), on the NANC nerve-mediated internal anal sphincter (IAS) smooth-muscle relaxation and the arginase activity in the IAS and other tissues. Arginase caused an attenuation of the IAS smooth-muscle relaxations by NANC nerve stimulation that was restored by the arginase inhibitors. L-HO-Arg but not ABH caused dose-dependent and complete reversal of N(omega)-nitro-L-arginine-suppressed IAS relaxation that was similar to that seen with L-arginine. Both ABH and L-HO-Arg caused an augmentation of NANC nerve-mediated relaxation of the IAS. In the IAS, ABH was found to be approximately 250 times more potent than L-HO-Arg in inhibiting the arginase activity. L-HO-Arg was found to be 10 to 18 times more potent in inhibiting the arginase activity in the liver than in nonhepatic tissues. We conclude that arginase plays a significant role in the regulation of nitric oxide synthase mediated NANC relaxation in the IAS. The advent of new and selective arginase inhibitors may play a significant role in the discrimination of arginase isozymes and have important pathophysiological and therapeutic implications in gastrointestinal motility disorders. PMID- 10454521 TI - Action potentials, contraction, and membrane currents in guinea pig ventricular preparations treated with the antispasmodic agent terodiline. AB - Terodiline was widely prescribed for urinary incontinence before reports of adverse cardiac effects that included bradycardia, QT lengthening, and ventricular tachyarrhythmia. The present study on guinea pig papillary muscles and ventricular myocytes was undertaken to gain insight into the cardioactive properties of the drug. Clinically relevant concentrations (<10 microM) of terodiline lengthened the action potential duration by up to 12%; higher concentrations shortened the duration in a concentration-dependent manner. The drug depressed maximal upstroke velocity in a use-dependent manner; the IC(50) value was near 150 microM in muscles driven at 1 Hz, 60 microM at 3 Hz, 38 microM at 5 Hz, and 3 microM at 1 Hz in muscles depolarized with 14 mM K(+). Submicromolar terodiline frequently had a small positive inotropic effect, whereas micromolar concentrations depressed force in a frequency-dependent manner. Voltage-clamp results on myocytes indicate that terodiline inhibits three membrane currents that govern repolarization: 1) E4031-sensitive, rapidly activating K(+) current with an IC(50) value near 0.7 microM as previously reported; 2) slowly activating, delayed-rectifier K(+) current with an IC(50) value of 26 microM; and 3) L-type Ca(2+) current with an IC(50) value of 12 microM. These findings are correlated with the changes in action potential configuration and developed tension and discussed in relation to the cardiotoxic effects of the drug. PMID- 10454522 TI - Differential effect of histamine 3 receptor-active agents on brain, but not peritoneal, mast cell activation. AB - The activation of presynaptic histamine 3 (H(3)) receptors inhibits the release of histamine and other neurotransmitters from central nervous system neurons. Rat brain mast cells (MCs) release histamine and 5-hydroxytryptamine (5-HT) in response to neuropeptides and neurotransmitters secreted from adjacent neurons. Dura MCs also degranulate in response to antidromic trigeminal nerve stimulation and with acute psychological stress. Such findings have implicated brain MCs in certain neuroinflammatory disorders, such as migraines. We investigated the ultrastructural appearance of control and stimulated thalamic/hypothalamic (brain) MCs before and after treatment with the H(3) receptor agonist N(alpha) methylhistamine (N(alpha)-mH) and the H(3) receptor antagonist thioperamide (Th). Ultrastructural investigation of brain MCs stimulated with compound 48/80 revealed extensive intragranular changes that paralleled 5-HT secretion but without degranulation by exocytosis typical of connective tissue MCs. N(alpha)-mH significantly reduced these morphological changes, as well as 5-HT release from brain MCs and neurons stimulated with KCl; conversely, Th augmented both histamine and 5-HT release from brain neurons and MCs. Neither N(alpha)-mH nor Th had any effect on peritoneal MCs. Simultaneous addition of both drugs largely antagonized each other's effects on brain MC activation and 5-HT secretion. Ultrastructural observations and lack of lactic dehydrogenase release in the perfusate excluded any cytotoxic effect. The ability of H(3) agonists to inhibit brain MC activation, as well as secretion of 5-HT from both brain MCs and neurons, may be useful in the management of migraines. PMID- 10454523 TI - Effect of calcium channel antagonists nifedipine and nicardipine on rat cytochrome P-450 2B and 3A forms. AB - Calcium channel antagonists are widely prescribed for treatment of hypertension. In this study, we examined whether treatment with the calcium channel antagonists, nicardipine, nifedipine or diltiazem, alters cytochrome P-450 2B or 3A (CYP2B or CYP3A, respectively) expression in rat liver. Western blot analyses were undertaken using antibodies specific for one or several members of these cytochrome P-450 subfamilies. Nicardipine was found to be an effective inducer of CYP3A; in particular, CYP3A23 was increased approximately 36-fold following treatment with 100 mg of nicardipine/kg/day. Nicardipine induced CYP2B forms up to approximately 3.1-fold. Nifedipine did not alter CYP3A expression but did increase CYP2B expression such that total CYP2B, CYP2B1, and CYP2B2v (a splice variant of CYP2B2) were increased approximately 5- to 15-fold after treatment with 100 mg of nifedipine/kg/day, with increases in benzyloxyresorufin O dealkylase and erythromycin N-demethylase activities, respectively. The distinct differences in cytochrome P-450 induction profile induced by nicardipine and nifedipine suggest that they may enhance cytochrome P-450 expression by different mechanisms unrelated to their effects on calcium channels. PMID- 10454524 TI - Tyrphostins that suppress the growth of human papilloma virus 16-immortalized human keratinocytes. AB - Human papilloma virus 16 (HPV16) is considered to be the causative agent for cervical cancer, which ranks second to breast cancer in women's malignancies. In an attempt to develop drugs that inhibit the malignant transformation of HPV16 immortalized epithelial cells, we examined the effect of tyrphostins on such cells. We examined the effect of tyrphostins from four different families on the growth of HPV16-immortalized human keratinocytes (HF-1) cells. We found that they alter their cell cycle distribution, their morphology, and induce cell death by apoptosis. The effects of tyrphostins on HF-1 cells are different from their effects on normal keratinocytes. Growth suppression by AG555 and AG1478 is accompanied by 30% apoptosis in HF-1 cells, but this is not observed in normal keratinocytes. Tyrphostin treatment produces distinctive morphological changes in HF-1 cells and in normal keratinocytes; however, the culture organization of normal keratinocytes is less disrupted. These differential effects of the tyrphostins on HPV16-immortalized keratinocytes compared with their effects on normal keratinocytes suggests that these compounds are suitable candidates for the treatment of papilloma. Previous and present results indicate that group 1 tyrphostins, which inhibit Cdk2 activation, and group 2 tyrphostins, represented by AG1478, a potent epidermal growth factor receptor kinase inhibitor, induce cell cycle arrest; and, in the case of HF-1 cells, apoptosis and differentiation. Cells accumulate in the G(1) phase of the cell cycle at the expense of S and G(2) + M. These compounds block the growth of normal keratinocytes without inducing apoptosis or differentiation, causing them to accumulate in G(1). AG17, which belongs to group 4, exerts its antiproliferative effect mainly by increasing the fractions of cells in G(1) with a concomitant decrease in the fraction of cells in S and G(2) + M. PMID- 10454525 TI - The human and rat recombinant receptors for advanced glycation end products have a high degree of homology but different pharmacokinetic properties in rats. AB - The accelerated formation of advanced glycation end products (AGEs) is implicated in diabetic microvascular and macrovascular complications. The binding of AGEs to their cellular surface receptor (RAGE) induces vascular dysfunction and in particular an increase in vascular permeability. We previously demonstrated that rat recombinant RAGE (rR-RAGE) produced in insect cells corrected the hyperpermeability due to RAGE-AGE interaction and that pharmacokinetic properties of rR-RAGE after i.v. administration in rats were compatible with a potential therapeutic use. In the present study, we showed that recombinant human RAGE (rH RAGE) had a similar efficacy in inhibiting AGE-induced endothelial alteration and in reducing the hyperpermeability observed in streptozotocin-induced diabetic rats. (125)I-rH-RAGE elimination half-life after i.v. administration was similar in diabetic and normal rats (53.7 +/- 7.6 and 45.3 +/- 4.0 h, respectively). The presence of AGEs is responsible for a higher distribution volume in diabetic rats compared with normal rats (15.3 +/- 2.7 and 7.7 +/- 0. 7 l/kg, respectively). Immunoreactive (125)I-rH-RAGE decreased more rapidly than did immunoreactive (125)I-rR-RAGE. The differences between (125)I-rH-RAGE and (125)I-rR-RAGE pharmacokinetics in rat may be related to differences in potential O glycosylation and protease cleavage sites between the two RAGE molecules. PMID- 10454526 TI - Retroviral-mediated expression of the P140A, but not P140A/G156A, mutant form of O6-methylguanine DNA methyltransferase protects hematopoietic cells against O6 benzylguanine sensitization to chloroethylnitrosourea treatment. AB - O(6)-Benzylguanine (6-BG) inactivates mammalian O(6)-methylguanine DNA methyltransferase (MGMT), an important DNA repair protein that protects cells against chloroethylnitrosourea (CENU) cytotoxicity. 6-BG is being tested as an approach to treat CENU-resistant tumors that overexpress endogenous MGMT. However, in addition to restoring CENU tumor cell sensitivity, 6-BG also increases the cytotoxic effects of CENUs on hematopoietic cells. Several 6-BG resistant human MGMT mutants have been characterized in Escherichia coli and are predicted to protect mammalian cells against the combination of 6-BG and CENU treatment in vivo. Two mutants, P140A and P140A/G156A, demonstrated 20- and 1200 fold more resistance to 6-BG depletion of MGMT activity compared with wild-type MGMT (WTMGMT). Here, we analyzed retroviral vectors that express either WTMGMT, the P140A or P140A/G156A mutant forms of MGMT. Retroviral-infected L1210 hematopoietic cells demonstrated similar levels of RNA in all transduced clones. However, the amount of MGMT protein and DNA repair activity was reduced in clones expressing the P140A/G156A mutant compared with those expressing WTMGMT or P140A. Expression of P140A was associated with a 4- to 8-fold increase in resistance to 6-BG depletion of MGMT in transduced L1210 clones and a 1, 3-bis(2-chloroethyl)-1 nitrosourea IC(50) of 50 microM (compared with 27.5 microM for WTMGMT) in primary murine hematopoietic cells. These results demonstrate the utility of screening 6 BG-resistant MGMT proteins in hematopoietic cells and provide evidence that the P140A mutant form of MGMT generates 6-BG- and CENU-resistant hematopoietic cells. Retrovirus vectors expressing this mutant may be useful in future human gene therapy trials. PMID- 10454527 TI - Inhibition of serotonin-induced vascular smooth muscle cell proliferation by sarpogrelate. AB - Antiproliferative behavior of sarpogrelate (Anplag, MCI-9042, (+/-)-1-[2-[2-(3 methoxyphenyl)ethyl]phenoxy]-3-(dimethylamino)-2-pro pyl hydrogen succinate hydrochloride), a serotonin 2A (5-HT2A) receptor antagonist, was established using radioactive incorporation of [(3)H]thymidine, [(3)H]uridine, and [(3)H]phenylalanine in cultured rat aortic smooth muscle cells in response to a 5 HT-induced cytokine trigger. Fluorescence-activated cell sorting was used to confirm these observations. 5-HT-induced DNA, RNA, and protein synthesis were inhibited maximally at a concentration of 1 microM sarpogrelate. Although other cytokines such as platelet-derived growth factor and endothelin also induced DNA, RNA, and protein synthesis in rat aortic smooth muscle cells, cell proliferation was not influenced by sarpogrelate, even at large pharmacological concentrations (10 microM). Sarpogrelate's antiproliferative actions were found to be more potent than ketanserin. These data indicate that sarpogrelate operates as a specific inhibitor of 5-HT-mediated cell proliferation and is a good candidate for preventing serotonin-induced neointimal hyperplasia. PMID- 10454528 TI - Functional characteristics and tissue distribution pattern of organic cation transporter 2 (OCTN2), an organic cation/carnitine transporter. AB - We have demonstrated in the present study that novel organic cation transporter (OCTN) 2 is a transporter for organic cations as well as carnitine. OCTN2 transports organic cations without involving Na(+), but it transports carnitine only in the presence of Na(+). The ability to transport organic cations and carnitine is demonstrable with human, rat, and mouse OCTN2s. Na(+) does not influence the affinity of OCTN2 for organic cations, but it increases the affinity severalfold for carnitine. The short-chain acyl esters of carnitine are also transported by OCTN2. Two mutations, M352R and P478L, in human OCTN2 are associated with loss of transport function, but the protein expression of these mutants is comparable to that of the wild-type human OCTN2. In situ hybridization in the rat shows that OCTN2 is expressed in the proximal and distal tubules and in the glomeruli in the kidney, in the myocardium, valves, and arterioles in the heart, in the labyrinthine layer of the placenta, and in the cortex, hippocampus, and cerebellum in the brain. This is the first report that OCTN2 is a Na(+) independent organic cation transporter as well as a Na(+)-dependent carnitine transporter and that OCTN2 is expressed not only in the heart, kidney, and placenta but also in the brain. PMID- 10454529 TI - Multisecond oscillations in firing rate in the globus pallidus: synergistic modulation by D1 and D2 dopamine receptors. AB - The firing rates of many basal ganglia neurons recorded in awake rats oscillate at seconds-to-minutes time scales, and the D1/D2 agonist apomorphine has been shown to robustly modulate these oscillations. The use of selective D1 and D2 antagonists suggested that both these receptor subfamilies are involved in apomorphine's effects. In the present study, spectral analysis revealed that baseline multisecond oscillations were significantly periodic in 71% of globus pallidus neurons. Baseline oscillations had a wide range of periods within the analyzed range, with a population mean of 32 +/- 2 s. Administration of the D1 agonist SKF 81297 (6-chloroPB) at 1.0 or 5.0 mg/kg significantly changed these oscillations, reducing means of spectral peak periods to 14 to 16 s (i.e., increasing oscillatory frequency). This effect was attenuated by D2 antagonist pretreatment. The D2 agonist quinpirole did not cause a significant population change in multisecond periodicities. The strongest effects on multisecond periodicities occurred after combined treatment with SKF 81297 and quinpirole. Low, ineffective doses of SKF 81297 and quinpirole, when combined, produced a significant increase in oscillatory frequency. Also, when quinpirole was administered after an already effective dose of SKF 81297, quinpirole shifted oscillations to an even faster range (typically to periods of <10 s). The dopaminergic control of multisecond periodicities in globus pallidus firing rate demonstrates D1/D2 receptor synergism, in that the effects of D1 agonists are potentiated by and partially dependent on D2 receptor activity. Modulation of multisecond oscillations in firing rate represents a novel means by which dopamine can influence globus pallidus physiology. PMID- 10454530 TI - Transcription and growth regulatory functions of the HIN-200 family of proteins. PMID- 10454531 TI - Predominance of duplicative VSG gene conversion in antigenic variation in African trypanosomes. AB - A number of mechanisms have been described by which African trypanosomes undergo the genetic switches that differentially activate their variant surface glycoprotein genes (VSGs) and bring about antigenic variation. These mechanisms have been observed mainly in trypanosome lines adapted, by rapid syringe passaging, to laboratory conditions. Such "monomorphic" lines, which routinely yield only the proliferative bloodstream form and do not develop through their life cycle, have VSG switch rates up to 4 or 5 orders of magnitude lower than those of nonadapted lines. We have proposed that nonadapted, or pleomorphic, trypanosomes normally have an active VSG switch mechanism, involving gene duplication, that is depressed, or from which a component is absent, in monomorphic lines. We have characterized 88 trypanosome clones from the first two relapse peaks of a single rabbit infection with pleomorphic trypanosomes and shown that they represent 11 different variable antigen types (VATs). The pattern of appearance in the first relapse peak was generally reproducible in three more rabbit infections. Nine of these VATs had activated VSGs by gene duplication, the tenth possibly also had done so, and only one had activated a VSG by the transcriptional switch mechanism that predominates in monomorphic lines. At least 10 of the donor genes have telomeric silent copies, and many reside on minichromosomes. It appears that trypanosome antigenic variation is dominated by one, relatively highly active, mechanism rather than by the plethora of pathways described before. PMID- 10454532 TI - Functional analysis of the SIN3-histone deacetylase RPD3-RbAp48-histone H4 connection in the Xenopus oocyte. AB - We investigated the protein associations and enzymatic requirements for the Xenopus histone deacetylase catalytic subunit RPD3 to direct transcriptional repression in Xenopus oocytes. Endogenous Xenopus RPD3 is present in nuclear and cytoplasmic pools, whereas RbAp48 and SIN3 are predominantly nuclear. We cloned Xenopus RbAp48 and SIN3 and show that expression of RPD3, but not RbAp48 or SIN3, leads to an increase in nuclear and cytoplasmic histone deacetylase activity and transcriptional repression of the TRbetaA promoter. This repression requires deacetylase activity and nuclear import of RPD3 mediated by a carboxy-terminal nuclear localization signal. Exogenous RPD3 is not incorporated into previously described oocyte deacetylase and ATPase complexes but cofractionates with a component of the endogenous RbAp48 in the oocyte nucleus. We show that RPD3 associates with RbAp48 through N- and C-terminal contacts and that RbAp48 also interacts with SIN3. Xenopus RbAp48 selectively binds to the segment of the N terminal tail immediately proximal to the histone fold domain of histone H4 in vivo. Exogenous RPD3 may be targeted to histones through interaction with endogenous RbAp48 to direct transcriptional repression of the Xenopus TRbetaA promoter in the oocyte nucleus. However, the exogenous RPD3 deacetylase functions to repress transcription in the absence of a requirement for association with SIN3 or other targeted corepressors. PMID- 10454533 TI - Dual role for Hsc70 in the biogenesis and regulation of the heme-regulated kinase of the alpha subunit of eukaryotic translation initiation factor 2. AB - The heme-regulated kinase of the alpha subunit of eukaryotic initiation factor 2 (HRI) is activated in rabbit reticulocyte lysate (RRL) in response to a number of environmental conditions, including heme deficiency, heat shock, and oxidative stress. Activation of HRI causes an arrest of initiation of protein synthesis. Recently, we have demonstrated that the heat shock cognate protein Hsc70 negatively modulates the activation of HRI in RRL in response to these environmental conditions. Hsc70 is also known to be a critical component of the Hsp90 chaperone machinery in RRL, which plays an obligatory role for HRI to acquire and maintain a conformation that is competent to activate. Using de novo synthesized HRI in synchronized pulse-chase translations, we have examined the role of Hsc70 in the regulation of HRI biogenesis and activation. Like Hsp90, Hsc70 interacted with nascent HRI and HRI that was matured to a state which was competent to undergo stimulus-induced activation (mature-competent HRI). Interaction of HRI with Hsc70 was required for the transformation of HRI, as the Hsc70 antagonist clofibric acid inhibited the folding of HRI into a mature competent conformation. Unlike Hsp90, Hsc70 also interacted with transformed HRI. Clofibric acid disrupted the interaction of Hsc70 with transformed HRI that had been matured and transformed in the absence of the drug. Disruption of Hsc70 interaction with transformed HRI in heme-deficient RRL resulted in its hyperactivation. Furthermore, activation of HRI in response to heat shock or denatured proteins also resulted in a similar blockage of Hsc70 interaction with transformed HRI. These results indicate that Hsc70 is required for the folding and transformation of HRI into an active kinase but is subsequently required to negatively attenuate the activation of transformed HRI. PMID- 10454534 TI - The G(2) checkpoint is maintained by redundant pathways. AB - While p53 activity is critical for a DNA damage-induced G(1) checkpoint, its role in the G(2) checkpoint has not been compelling because cells lacking p53 retain the ability to arrest in G(2) following DNA damage. Comparison between normal human foreskin fibroblasts (HFFs) and HFFs in which p53 was eliminated by transduction with human papillomavirus type 16 E6 showed that treatment with adriamycin initiated arrest in G(2) with active cyclin B/CDC2 kinase, regardless of p53 status. Both E6-transduced HFFs and control (LXSN)-transduced cells maintained a prolonged arrest in G(2); however cells with functional p53 extinguished cyclin B-associated kinase activity. Down regulation was mediated by p53-dependent transcriptional repression of the CDC2 and cyclin B promoters. In contrast, cells lacking p53 showed a prolonged G(2) arrest despite high levels of cyclin B/CDC2 kinase activity, at least some of which translocated into the nucleus. Furthermore, the G(2) checkpoint became attenuated as p53-deficient cells aged in culture. Thus, at late passage, E6-transduced HFFs entered mitosis following DNA damage, whereas the age-matched parental HFFs sustained a G(2) arrest. These results indicate that normal cells have p53-independent pathways to maintain DNA damage-induced G(2) arrest, which may be augmented by p53-dependent functions, and that cells lacking p53 are at greater risk of losing the pathway that protects against aneuploidy. PMID- 10454535 TI - Protein kinase A-dependent and -independent signaling pathways contribute to cyclic AMP-stimulated proliferation. AB - The effects of cyclic AMP (cAMP) on cell proliferation are cell type specific. Although the growth-inhibitory effects of cAMP have been well studied, much less is known regarding how cAMP stimulates proliferation. We report that cAMP stimulates proliferation through both protein kinase A (PKA)-dependent and PKA independent signaling pathways and that phosphatidylinositol 3-kinase (PI3K) is required for cAMP-stimulated mitogenesis. In cells where cAMP is a mitogen, cAMP elevating agents stimulate membrane ruffling, Akt phosphorylation, and p70 ribosomal S6 protein kinase (p70s6k) activity. cAMP effects on ruffle formation and Akt were PKA independent but sensitive to wortmannin. In contrast, cAMP stimulated p70s6k activity was repressed by PKA inhibitors but not by wortmannin or microinjection of the N-terminal SH2 domain of the p85 regulatory subunit of PI3K, indicating that p70s6k and Akt can be regulated independently. Microinjection of highly specific inhibitors of PI3K or Rac1, or treatment with the p70s6k inhibitor rapamycin, impaired cAMP-stimulated DNA synthesis, demonstrating that PKA-dependent and -independent pathways contribute to cAMP mediated mitogenesis. Direct elevation of PI3K activity through microinjection of an antibody that stimulates PI3K activity or stable expression of membrane localized p110 was sufficient to confer hormone-independent DNA synthesis when accompanied by elevations in p70s6k activity. These findings indicate that multiple pathways contribute to cAMP-stimulated mitogenesis, only some of which are PKA dependent. Furthermore, they demonstrate that the ability of cAMP to stimulate both p70s6k- and PI3K-dependent pathways is an important facet of cAMP regulated cell cycle progression. PMID- 10454536 TI - Suppression of Ras-induced apoptosis by the Rac GTPase. AB - Ras is an essential component of signal transduction pathways that control cell proliferation, differentiation, and survival. In this study we have examined the cellular responses to high-intensity Ras signaling. Expression of increasing amounts of the oncogenic form of human HRas, HRasV12, results in a dose-dependent induction of apoptosis in both primary and immortalized cells. The induction of apoptosis by HRasV12 is blocked by activated Rac and potentiated by dominant interfering Rac. The ability of Rac to suppress Ras-induced apoptosis is dependent on effector pathway(s) controlled by the insert region and is linked to the activation of NF-kappaB. The apoptotic effect of HRasV12 requires the activation of both the ERK and JNK mitogen-activated protein kinase cascade and is independent of p53. These results demonstrate a role for Rac in controlling signals that are necessary for cell survival, and suggest a mechanism by which Rac activity can confer growth advantage to cells transformed by the ras oncogene. PMID- 10454537 TI - The von Hippel-Lindau tumor suppressor gene inhibits hepatocyte growth factor/scatter factor-induced invasion and branching morphogenesis in renal carcinoma cells. AB - Loss of function in the von Hippel-Lindau (VHL) tumor suppressor gene occurs in familial and most sporadic renal cell carcinomas (RCCs). VHL has been linked to the regulation of cell cycle cessation (G(0)) and to control of expression of various mRNAs such as for vascular endothelial growth factor. RCC cells express the Met receptor tyrosine kinase, and Met mediates invasion and branching morphogenesis in many cell types in response to hepatocyte growth factor/scatter factor (HGF/SF). We examined the HGF/SF responsiveness of RCC cells containing endogenous mutated (mut) forms of the VHL protein (VHL-negative RCC) with that of isogenic cells expressing exogenous wild-type (wt) VHL (VHL-positive RCC). We found that VHL-negative 786-0 and UOK-101 RCC cells were highly invasive through growth factor-reduced (GFR) Matrigel-coated filters and exhibited an extensive branching morphogenesis phenotype in response to HGF/SF in the three-dimensional (3D) GFR Matrigel cultures. In contrast, the phenotypes of A498 VHL-negative RCC cells were weaker, and isogenic RCC cells ectopically expressing wt VHL did not respond at all. We found that all VHL-negative RCC cells expressed reduced levels of tissue inhibitor of metalloproteinase 2 (TIMP-2) relative to the wt VHL positive cells, implicating VHL in the regulation of this molecule. However, consistent with the more invasive phenotype of the 786-0 and UOK-101 VHL-negative RCC cells, the levels of TIMP-1 and TIMP-2 were reduced and levels of the matrix metalloproteinases 2 and 9 were elevated compared to the noninvasive VHL-positive RCC cells. Moreover, recombinant TIMPs completely blocked HGF/SF-mediated branching morphogenesis, while neutralizing antibodies to the TIMPs stimulated HGF/SF-mediated invasion in vitro. Thus, the loss of the VHL tumor suppressor gene is central to changes that control tissue invasiveness, and a more invasive phenotype requires additional genetic changes seen in some but not all RCC lines. These studies also demonstrate a synergy between the loss of VHL function and Met signaling. PMID- 10454538 TI - Myc downregulation by transforming growth factor beta required for activation of the p15(Ink4b) G(1) arrest pathway. AB - The antimitogenic action of transforming growth factor beta (TGF-beta) in epithelial cells involves cyclin-dependent kinase (cdk) inhibitory gene responses and downregulation of c-Myc expression. Although the cdk inhibitory responses are sufficient for G(1) arrest, enforced expression of c-Myc prevents G(1) arrest by TGF-beta. We investigated the basis of this antagonism by using Mv1Lu lung epithelial cell lines that conditionally express levels of human c-Myc. We show that c-Myc prevents induction of the cdk4 inhibitor p15(Ink4b) and the subsequent inhibition of G(1) cdks by TGF-beta. We assessed the significance of this effect by analyzing the oligomeric state of cdk4 in these cells. In proliferating cells, endogenous cdk4 is distributed among three populations: an abundant high molecular-mass (>400-kDa) pool of latent cdk4 that serves as a source of cdk4 for cyclin D, a low-abundance pool containing active cyclin D-cdk4 complexes, and an inactive population of monomeric cdk4. Cell stimulation with TGF-beta converts the latent and active cdk4 pools into inactive cdk4, an effect that is specifically mimicked by overexpression of p15 but not by other forms of G(1) arrest. This process of TGF-beta-induced cdk4 inactivation is completely blocked by expression of c-Myc, even though the latent and active cdk4 complexes from c Myc-expressing cells remain sensitive to dissociation by p15 in vitro. c-Myc causes a small increase in cyclin D levels, but this effect contributes little to the loss of TGF-beta responses in these cells. The evidence suggests that c-Myc interferes with TGF-beta activation of the p15 G(1) arrest pathway. TGF-beta must therefore downregulate c-Myc in order to activate this pathway. PMID- 10454539 TI - NF-kappaB induces expression of the Bcl-2 homologue A1/Bfl-1 to preferentially suppress chemotherapy-induced apoptosis. AB - Recent evidence indicates that the transcription factor NF-kappaB is a major effector of inducible antiapoptotic mechanisms. For example, it was shown that NF kappaB activation suppresses the activation of caspase 8, the apical caspase in tumor necrosis factor (TNF) receptor family signaling cascades, through the transcriptional regulation of certain TRAF and IAP proteins. However, it was unknown whether NF-kappaB controls other key regulatory mechanisms in apoptosis. Here we show that NF-kappaB activation suppresses mitochondrial release of cytochrome c through the activation of the Bcl-2 family member A1/Bfl-1. The restoration of A1 in NF-kappaB null cells diminished TNF-induced apoptosis by reducing the release of proapoptotic cytochrome c from mitochondria. In addition, A1 potently inhibited etoposide-induced apoptosis by inhibiting the release of cytochrome c and by blocking caspase 3 activation. Our findings demonstrate that A1 is an important antiapoptotic gene controlled by NF-kappaB and establish that the prosurvival function of NF-kappaB can be manifested at multiple levels. PMID- 10454540 TI - Active-site mutations in the Xrn1p exoribonuclease of Saccharomyces cerevisiae reveal a specific role in meiosis. AB - Xrn1p of Saccharomyces cerevisiae is a major cytoplasmic RNA turnover exonuclease which is evolutionarily conserved from yeasts to mammals. Deletion of the XRN1 gene causes pleiotropic phenotypes, which have been interpreted as indirect consequences of the RNA turnover defect. By sequence comparisons, we have identified three loosely defined, common 5'-3' exonuclease motifs. The significance of motif II has been confirmed by mutant analysis with Xrn1p. The amino acid changes D206A and D208A abolish singly or in combination the exonuclease activity in vivo. These mutations show separation of function. They cause identical phenotypes to that of xrn1Delta in vegetative cells but do not exhibit the severe meiotic arrest and the spore lethality phenotype typical for the deletion. In addition, xrn1-D208A does not cause the severe reduction in meiotic popout recombination in a double mutant with dmc1 as does xrn1Delta. Biochemical analysis of the DNA binding, exonuclease, and homologous pairing activity of purified mutant enzyme demonstrated the specific loss of exonuclease activity. However, the mutant enzyme is competent to promote in vitro assembly of tubulin into microtubules. These results define a separable and specific function of Xrn1p in meiosis which appears unrelated to its RNA turnover function in vegetative cells. PMID- 10454541 TI - SMG-2 is a phosphorylated protein required for mRNA surveillance in Caenorhabditis elegans and related to Upf1p of yeast. AB - mRNAs that contain premature stop codons are selectively degraded in all eukaryotes tested, a phenomenon termed "nonsense-mediated mRNA decay" (NMD) or "mRNA surveillance." NMD may function to eliminate aberrant mRNAs so that they are not translated, because such mRNAs might encode deleterious polypeptide fragments. In both yeasts and nematodes, NMD is a nonessential system. Mutations affecting three yeast UPF genes or seven nematode smg genes eliminate NMD. We report here the molecular analysis of smg-2 of Caenorhabditis elegans. smg-2 is homologous to UPF1 of yeast and to RENT1 (also called HUPF1), a human gene likely involved in NMD. The striking conservation of SMG-2, Upf1p, and RENT1/HUPF1 in both sequence and function suggests that NMD is an ancient system, predating the divergence of most eukaryotes. Despite similarities in the sequences of SMG-2 and Upf1p, expression of Upf1p in C. elegans does not rescue smg-2 mutants. We have prepared anti-SMG-2 polyclonal antibodies and identified SMG-2 on Western blots. SMG-2 is phosphorylated, and mutations of the six other smg genes influence the state of SMG-2 phosphorylation. In smg-1, smg-3, and smg-4 mutants, phosphorylation of SMG-2 was not detected. In smg-5, smg-6, and smg-7 mutants, a phosphorylated isoform of SMG-2 accumulated to abnormally high levels. In smg 2(r866) and smg-2(r895) mutants, which harbor single amino acid substitutions of the SMG-2 nucleotide binding site, phosphorylated SMG-2 accumulated to abnormally high levels, similar to those observed in smg-5, smg-6, and smg-7 mutants. We discuss these results with regard to the in vivo functions of SMG-2 and NMD. PMID- 10454542 TI - Histone acetyltransferase complexes can mediate transcriptional activation by the major glucocorticoid receptor activation domain. AB - Previous studies have shown that the Ada adapter proteins are important for glucocorticoid receptor (GR)-mediated gene activation in yeast. The N-terminal transactivation domain of GR, tau1, is dependent upon Ada2, Ada3, and Gcn5 for transactivation in vitro and in vivo. Using in vitro techniques, we demonstrate that the GR-tau1 interacts directly with the native Ada containing histone acetyltransferase (HAT) complex SAGA but not the related Ada complex. Mutations in tau1 that reduce tau1 transactivation activity in vivo lead to a reduced binding of tau1 to the SAGA complex and conversely, mutations increasing the transactivation activity of tau1 lead to an increased binding of tau1 to SAGA. In addition, the Ada-independent NuA4 HAT complex also interacts with tau1. GAL4 tau1-driven transcription from chromatin templates is stimulated by SAGA and NuA4 in an acetyl coenzyme A-dependent manner. Low-activity tau1 mutants reduce SAGA- and NuA4-stimulated transcription while high-activity tau1 mutants increase transcriptional activation, specifically from chromatin templates. Our results demonstrate that the targeting of native HAT complexes by the GR-tau1 activation domain mediates transcriptional stimulation from chromatin templates. PMID- 10454543 TI - Tat-SF1 protein associates with RAP30 and human SPT5 proteins. AB - The potent transactivator Tat recognizes the transactivation response RNA element (TAR) of human immunodeficiency virus type 1 and stimulates the processivity of elongation of RNA polymerase (Pol) II complexes. The cellular proteins Tat-SF1 and human SPT5 (hSPT5) are required for Tat activation as shown by immunodepletion with specific sera and complementation with recombinant proteins. In nuclear extracts, small fractions of both hSPT5 and Pol II are associated with Tat-SF1 protein. Surprisingly, the RAP30 protein of the heterodimeric transcription TFIIF factor is associated with Tat-SF1, while the RAP74 subunit of TFIIF is not coimmunoprecipitated with Tat-SF1. Overexpression of Tat-SF1 and hSPT5 specifically stimulates the transcriptional activity of Tat in vivo. These results suggest that Tat-SF1 and hSPT5 are indispensable cellular factors supporting Tat-specific transcription activation and that they may interact with RAP30 in controlling elongation. PMID- 10454545 TI - Phosphorylation-independent inhibition of Cdc28p by the tyrosine kinase Swe1p in the morphogenesis checkpoint. AB - The morphogenesis checkpoint in budding yeast delays cell cycle progression in G(2) when the actin cytoskeleton is perturbed, providing time for cells to complete bud formation prior to mitosis. Checkpoint-induced G(2) arrest involves the inhibition of the master cell cycle regulatory cyclin-dependent kinase, Cdc28p, by the Wee1 family kinase Swe1p. Results of experiments using a nonphosphorylatable CDC28(Y19F) allele suggested that the checkpoint stimulated two inhibitory pathways, one that promoted phosphorylation at tyrosine 19 (Y19) and a poorly characterized second pathway that did not require Cdc28p Y19 phosphorylation. We present the results from a genetic screen for checkpoint defective mutants that led to the repeated isolation of the dominant CDC28(E12K) allele that is resistant to Swe1p-mediated inhibition. Comparison of this allele with the nonphosphorylatable CDC28(Y19F) allele suggested that Swe1p is still able to inhibit CDC28(Y19F) in a phosphorylation-independent manner and that both the Y19 phosphorylation-dependent and -independent checkpoint pathways in fact reflect Swe1p inhibition of Cdc28p. Remarkably, we found that a Swe1p mutant lacking catalytic activity could significantly delay the cell cycle in vivo during a physiological checkpoint response, even when expressed at single copy. The finding that a Wee1 family kinase expressed at physiological levels can inhibit a nonphosphorylatable cyclin-dependent kinase has broad implications for many checkpoint studies using such mutants in other organisms. PMID- 10454544 TI - Impaired immune responses and B-cell proliferation in mice lacking the Id3 gene. AB - B-lymphocyte activation and proliferation induced by the B-cell receptor (BCR) signals are important steps in the initiation of humoral immune responses. How the BCR signals are translated by nuclear transcription factors into cell cycle progression is poorly understood. Id3 is an immediate-early gene responding to growth and mitogenic signals in many cell types including B cells. The primary function of the Id3 protein has been defined as that of inhibitor of basic-helix loop-helix (bHLH) transcription factors. The interaction between Id3 and bHLH proteins, many of which are essential for cellular differentiation, has been proposed as a key regulatory event leading to cellular proliferation instead of differentiation. To further investigate the role of Id3 in tissue and embryo development and the mechanism of Id3-mediated growth regulation, we generated and analyzed Id3-deficient mice. While these mice display no overt abnormality in tissue and embryo development, their humoral immunity is compromised. The amounts of immunoglobulins produced in Id3-deficient mice immunized with a T-cell dependent antigen and a type 2 T-cell-independent antigen are attenuated and severely impaired, respectively. Further analysis of lymphocytes isolated from Id3-deficient mice reveals a B-cell defect in their proliferation response to BCR cross-linking but not to lipopolysaccharide or a combination of BCR cross-linking and interleukin-4. Analyses of cultured lymphocytes also suggest involvement of Id3 in cytokine production in T cells and isotype switching in B cells. Finally, the proliferation defect in Id3-deficient B cells can be rescued by ectopic expression of Id1, a homologue of Id3. Taken together, these results define a necessary and specific role for Id3 in mediating signals from BCR to cell cycle progression during humoral immune responses. PMID- 10454546 TI - Inhibition of mitogen-activated kinase signaling sensitizes HeLa cells to Fas receptor-mediated apoptosis. AB - The Fas receptor (FasR) is an important physiological mediator of apoptosis in various tissues and cells. However, there are also many FasR-expressing cell types that are normally resistant to apoptotic signaling through this receptor. The mitogen-activated protein kinase (MAPK) signaling cascade has, apart from being a growth-stimulating factor, lately received attention as an inhibitory factor in apoptosis. In this study, we examined whether MAPK signaling could be involved in protecting FasR-insensitive cells. To this end, we used different approaches to inhibit MAPK signaling in HeLa cells, including treatment with the MAPK kinase inhibitor PD 98059, serum withdrawal, and expression of dominant interfering MAPK kinase mutant protein. All of these treatments were effective in sensitizing the cells to FasR-induced apoptosis, demonstrating that MAPK indeed is involved in the control of FasR responses. The MAPK-mediated control seemed to occur at or upstream of caspase 8, the initiator caspase in apoptotic FasR responses. Transfection with the constitutively active MAPK kinase abrogated FasR induced apoptosis also in the presence of cycloheximide, indicating that the MAPK generated suppression of FasR-mediated apoptotic signaling is protein synthesis independent. In cells insensitive to FasR-induced apoptosis, stimulation of the FasR with an agonistic antibody resulted in significant MAPK activation, which was inhibited by PD 98059. When different cell types were compared, the FasR mediated MAPK activation seemed proportional to the degree of FasR insensitivity. These results suggest that the FasR insensitivity is likely to be a consequence of FasR-induced MAPK activation, which in turn interferes with caspase activation. PMID- 10454548 TI - Base pairing between U3 small nucleolar RNA and the 5' end of 18S rRNA is required for pre-rRNA processing. AB - The loop of a stem structure close to the 5' end of the 18S rRNA is complementary to the box A region of the U3 small nucleolar RNA (snoRNA). Substitution of the 18S loop nucleotides inhibited pre-rRNA cleavage at site A(1), the 5' end of the 18S rRNA, and at site A(2), located 1.9 kb away in internal transcribed spacer 1. This inhibition was largely suppressed by a compensatory mutation in U3, demonstrating functional base pairing. The U3-pre-rRNA base pairing is incompatible with the structure that forms in the mature 18S rRNA and may prevent premature folding of the pre-rRNA. In the Escherichia coli pre-rRNA the homologous region of the 16S rRNA is also sequestered, in that case by base pairing to the 5' external transcribed spacer (5' ETS). Cleavage at site A(0) in the yeast 5' ETS strictly requires base pairing between U3 and a sequence within the 5' ETS. In contrast, the U3-18S interaction is not required for A(0) cleavage. U3 therefore carries out at least two functionally distinct base pair interactions with the pre-rRNA. The nucleotide at the site of A(1) cleavage was shown to be specified by two distinct signals; one of these is the stem-loop structure within the 18S rRNA. However, in contrast to the efficiency of cleavage, the position of A(1) cleavage is not dependent on the U3-loop interaction. We conclude that the 18S stem-loop structure is recognized at least twice during pre-rRNA processing. PMID- 10454549 TI - Targeted expression of the DNA binding domain of DRE-binding factor, a Drosophila transcription factor, attenuates DNA replication of the salivary gland and eye imaginal disc. AB - The promoters of Drosophila genes encoding DNA replication-related proteins contain transcription regulatory elements consisting of an 8-bp palindromic DNA replication-related element (DRE) sequence (5'-TATCGATA). The specific DRE binding factor (DREF), a homodimer of the polypeptide with 709 amino acid residues, is a positive trans-acting factor for transcription of DRE-containing genes. Both DRE binding and dimer formation are associated with residues 16 to 115 of the N-terminal region. We have established transgenic flies expressing the full-length DREF polypeptide or its N-terminal fragment (amino acid residues 1 to 125) under the control of the heat shock promoter, the salivary gland-specific promoter, or the eye imaginal disc-specific promoter. Heat shock induction of the N-terminal fragment during embryonic, larval, or pupal stages caused greater than 50% lethality. This lethality was overcome by coexpression of the full-length DREF. In salivary glands of the transgenic larvae expressing the N-terminal fragment, this fragment formed a homodimer and a heterodimer with the endogenous DREF. Ectopic expression of the N-terminal fragment in salivary gland cells reduced the contents of mRNAs for the 180-kDa subunit of DNA polymerase alpha and for dE2F and the extent of DNA endoreplication. Ectopic expression of the N terminal fragment in the eye imaginal discs significantly reduced DNA replication in cells at the second mitotic wave. The lines of evidence suggest that the N terminal fragment can impede the endogenous DREF function in a dominant negative manner and that DREF is required for normal DNA replication in both mitotic cell cycle and endo cycle. PMID- 10454547 TI - The mitogen-activated protein kinase kinase/extracellular signal-regulated kinase cascade activation is a key signalling pathway involved in the regulation of G(1) phase progression in proliferating hepatocytes. AB - In this study, activation of the mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signalling pathway was analyzed in proliferating rat hepatocytes both in vivo after partial hepatectomy and in vitro following epidermal growth factor (EGF)-pyruvate stimulation. First, a biphasic MEK/ERK activation was evidenced in G(1) phase of hepatocytes from regenerating liver but not from sham-operated control animals. One occurred in early G(1) (30 min to 4 h), and the other occurred in mid-late G(1), peaking at around 10.5 h. Interestingly, the mid-late G(1) activation peak was located just before cyclin D1 induction in both in vivo and in vitro models. Second, the biological role of the MEK/ERK cascade activation in hepatocyte progression through the G(1)/S transition was assessed by adding a MEK inhibitor (PD 98059) to EGF-pyruvate-stimulated hepatocytes in primary culture. In the presence of MEK inhibitor, cyclin D1 mRNA accumulation was inhibited, DNA replication was totally abolished, and the MEK1 isoform was preferentially targeted by this inhibition. This effect was dose dependent and completely reversed by removing the MEK inhibitor. Furthermore, transient transfection of hepatocytes with activated MEK1 construct resulted in increased cyclin D1 mRNA accumulation. Third, a correlation between the mid-late G(1) MEK/ERK activation in hepatocytes in vivo after partial hepatectomy and the mitogen-independent proliferation capacity of these cells in vitro was established. Among hepatocytes isolated either 5, 7, 9, 12 or 15 h after partial hepatectomy, only those isolated from 12- and 15-h regenerating livers were able to replicate DNA without additional growth stimulation in vitro. In addition, PD 98059 intravenous administration in vivo, before MEK activation, was able to inhibit DNA replication in hepatocytes from regenerating livers. Taken together, these results show that (i) early induction of the MEK/ERK cascade is restricted to hepatocytes from hepatectomized animals, allowing an early distinction of primed hepatocytes from those returning to quiescence, and (ii) mid-late G(1) MEK/ERK activation is mainly associated with cyclin D1 accumulation which leads to mitogen-independent progression of hepatocytes to S phase. These results allow us to point to a growth factor dependency in mid-late G(1) phase of proliferating hepatocytes in vivo as observed in vitro in proliferating hepatocytes and argue for a crucial role of the MEK/ERK cascade signalling pathway. PMID- 10454550 TI - Functional analysis of the yeast Glc7-binding protein Reg1 identifies a protein phosphatase type 1-binding motif as essential for repression of ADH2 expression. AB - In Saccharomyces cerevisiae, the protein phosphatase type 1 (PP1)-binding protein Reg1 is required to maintain complete repression of ADH2 expression during growth on glucose. Surprisingly, however, mutant forms of the yeast PP1 homologue Glc7, which are unable to repress expression of another glucose-regulated gene, SUC2, fully repressed ADH2. Constitutive ADH2 expression in reg1 mutant cells did require Snf1 protein kinase activity like constitutive SUC2 expression and was inhibited by unregulated cyclic AMP-dependent protein kinase activity like ADH2 expression in derepressed cells. To further elucidate the functional role of Reg1 in repressing ADH2 expression, deletions scanning the entire length of the protein were analyzed. Only the central region of the protein containing the putative PP1-binding sequence RHIHF was found to be indispensable for repression. Introduction of the I466M F468A substitutions into this sequence rendered Reg1 almost nonfunctional. Deletion of the central region or the double substitution prevented Reg1 from significantly interacting with Glc7 in two-hybrid analyses. Previous experimental evidence had indicated that Reg1 might target Glc7 to nuclear substrates such as the Snf1 kinase complex. Subcellular localization of a fully functional Reg1-green fluorescent protein fusion, however, indicated that Reg1 is cytoplasmic and excluded from the nucleus independently of the carbon source. When the level of Adr1 was modestly elevated, ADH2 expression was no longer fully repressed in glc7 mutant cells, providing the first direct evidence that Glc7 can repress ADH2 expression. These results suggest that the Reg1-Glc7 phosphatase is a cytoplasmic component of the machinery responsible for returning Snf1 kinase activity to its basal level and reestablishing glucose repression. This implies that the activated form of the Snf1 kinase complex must cycle between the nucleus and the cytoplasm. PMID- 10454551 TI - Cell cycle progression and proliferation despite 4BP-1 dephosphorylation. AB - Proliferation and cell cycle progression in response to growth factors require de novo protein synthesis. It has been proposed that binding of the eukaryotic translation initiation factor 4E (eIF-4E) to the inhibitory protein 4BP-1 blocks translation by preventing access of eIF-4G to the 5' cap of the mRNA. The signal for translation initiation is thought to involve phosphorylation of 4BP-1, which causes it to dissociate from eIF-4E and allows eIF-4G to localize to the 5' cap. It has been suggested that the ability of the macrolide antibiotic rapamycin to inhibit 4BP-1 phosphorylation is responsible for the potent antiproliferative property of this drug. We now show that rapamycin-resistant cells exhibited normal proliferation despite dephosphorylation of 4BP-1 that allows it to bind to eIF-4E. Moreover, despite rapamycin-induced dephosphorylation of 4BP-1, eIF-4E eIF-4G complexes (eIF-4F) were still detected. In contrast, amino acid withdrawal, which caused a similar degree of 4BP-1 dephosphorylation, resulted in dissociation of the eIF-4E-eIF-4G complex. Thus, 4BP-1 dephosphorylation is not equivalent to eIF-4E inactivation and does not explain the antiproliferative property of rapamycin. PMID- 10454552 TI - E2F4 actively promotes the initiation and maintenance of nerve growth factor induced cell differentiation. AB - E2F transcription factors play a critical role in cell cycle progression through the regulation of genes required for G(1)/S transition. They are also thought to be important for growth arrest; however, their potential role in the cell differentiation process has not been previously examined. Here, we demonstrate that E2F4 is highly upregulated following the neuronal differentiation of PC12 cells with nerve growth factor (NGF), while E2F1, E2F3, and E2F5 are downregulated. Immunoprecipitation and subcellular fractionation studies demonstrated that both the nuclear localization of E2F4 and its association with the Rb family member p130 increased following neuronal differentiation. The forced expression of E2F4 markedly enhanced the rate of PC12 cell differentiation induced by NGF and also greatly lowered the rate at which cells lost their neuronal phenotype following NGF removal. Importantly, this effect occurred in the absence of any significant change in the growth regulation of PC12 cells by NGF. Further, the downregulation of E2F4 expression with antisense oligodeoxynucleotides inhibited NGF-induced neurite outgrowth, indicating an important role for this factor during PC12 cell differentiation. Finally, E2F4 expression was found to increase dramatically in the developing rat cerebral cortex and cerebellum, as neuroblasts became postmitotic and initiated terminal differentiation. These findings demonstrate that, in addition to its effects on cell proliferation, E2F4 actively promotes the neuronal differentiation of PC12 cells as well as the retention of this state. Further, this effect is independent of alterations in cell growth and may involve interactions between E2F4 and the neuronal differentiation program itself. E2F4 may be an important participant in the terminal differentiation of neuroblasts. PMID- 10454553 TI - The strength of interaction at the Raf cysteine-rich domain is a critical determinant of response of Raf to Ras family small GTPases. AB - To be fully activated at the plasma membrane, Raf-1 must establish two distinct modes of interactions with Ras, one through its Ras-binding domain and the other through its cysteine-rich domain (CRD). The Ras homologue Rap1A is incapable of activating Raf-1 and even antagonizes Ras-dependent activation of Raf-1. We proposed previously that this property of Rap1A may be attributable to its greatly enhanced interaction with Raf-1 CRD compared to Ras. On the other hand, B Raf, another Raf family member, is activatable by both Ras and Rap1A. When interactions with Ras and Rap1A were measured, B-Raf CRD did not exhibit the enhanced interaction with Rap1A, suggesting that the strength of interaction at CRDs may account for the differential action of Rap1A on Raf-1 and B-Raf. The importance of the interaction at the CRD is further supported by a domain shuffling experiment between Raf-1 and B-Raf, which clearly indicated that the nature of CRD determines the specificity of response to Rap1A: Raf-1, whose CRD is replaced by B-Raf CRD, became activatable by Rap1A, whereas B-Raf, whose CRD is replaced by Raf-1 CRD, lost its response to Rap1A. Finally, a B-Raf CRD mutant whose interaction with Rap1A is selectively enhanced was isolated and found to possess the double mutation K252E/M278T. B-Raf carrying this mutation was not activated by Rap1A but retained its response to Ras. These results indicate that the strength of interaction with Ras and Rap1A at its CRD may be a critical determinant of regulation of the Raf kinase activity by the Ras family small GTPases. PMID- 10454554 TI - Interactions of TLC1 (which encodes the RNA subunit of telomerase), TEL1, and MEC1 in regulating telomere length in the yeast Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, chromosomes terminate with a repetitive sequence [poly(TG(1-3))] 350 to 500 bp in length. Strains with a mutation of TEL1, a homolog of the human gene (ATM) mutated in patients with ataxia telangiectasia, have short but stable telomeric repeats. Mutations of TLC1 (encoding the RNA subunit of telomerase) result in strains that have continually shortening telomeres and a gradual loss of cell viability; survivors of senescence arise as a consequence of a Rad52p-dependent recombination events that amplify telomeric and subtelomeric repeats. We show that a mutation in MEC1 (a gene related in sequence to TEL1 and ATM) reduces telomere length and that tel1 mec1 double mutant strains have a senescent phenotype similar to that found in tlc1 strains. As observed in tlc1 strains, survivors of senescence in the tel1 mec1 strains occur by a Rad52p-dependent amplification of telomeric and subtelomeric repeats. In addition, we find that strains with both tel1 and tlc1 mutations have a delayed loss of cell viability compared to strains with the single tlc1 mutation. This result argues that the role of Tel1p in telomere maintenance is not solely a direct activation of telomerase. PMID- 10454555 TI - Cleavage and inactivation of ATM during apoptosis. AB - The activation of the cysteine proteases with aspartate specificity, termed caspases, is of fundamental importance for the execution of programmed cell death. These proteases are highly specific in their action and activate or inhibit a variety of key protein molecules in the cell. Here, we study the effect of apoptosis on the integrity of two proteins that have critical roles in DNA damage signalling, cell cycle checkpoint controls, and genome maintenance-the product of the gene defective in ataxia telangiectasia, ATM, and the related protein ATR. We find that ATM but not ATR is specifically cleaved in cells induced to undergo apoptosis by a variety of stimuli. We establish that ATM cleavage in vivo is dependent on caspases, reveal that ATM is an efficient substrate for caspase 3 but not caspase 6 in vitro, and show that the in vitro caspase 3 cleavage pattern mirrors that in cells undergoing apoptosis. Strikingly, apoptotic cleavage of ATM in vivo abrogates its protein kinase activity against p53 but has no apparent effect on the DNA binding properties of ATM. These data suggest that the cleavage of ATM during apoptosis generates a kinase-inactive protein that acts, through its DNA binding ability, in a trans dominant-negative fashion to prevent DNA repair and DNA damage signalling. PMID- 10454556 TI - The androgen receptor amino-terminal domain plays a key role in p160 coactivator stimulated gene transcription. AB - Steroid receptors are conditional transcription factors that, upon binding to their response elements, regulate the expression of target genes via direct protein interactions with transcriptional coactivators. We have analyzed the functional interactions between the androgen receptor (AR) and 160-kDa nuclear receptor coactivators. Upon overexpression in mammalian cells, these coactivators enhance the transcriptional activity of both the amino-terminal domain (NTD) and the ligand-binding domain (LBD) of the AR. The coactivator activity for the LBD is strictly ligand-controlled and depends on the nature of the DNA-binding domain to which it is fused. We demonstrate that the NTD physically interacts with coactivators and with the LBD and that this interaction, like the functional interaction between the LBD and p160 coactivators, relies on the activation function 2 (AF2) core domain. The mutation of a highly conserved lysine residue in the predicted helix 3 of the LBD (K720A), however, blunts the functional interaction with coactivators but not with the NTD. Moreover, this mutation does not affect the transcriptional activity of the full-size AR. A mutation in the NTD of activation function AF1a (I182A/L183A), which dramatically impairs the activity of the AR, has no effect on the intrinsic transcriptional activity of the NTD but interferes with the cooperation between the NTD and the LBD. Finally, p160 proteins in which the three LXXLL motifs are mutated retain most of their coactivator activity for the full-size AR, although they are no longer functional for the isolated LBD. Together, these data suggest that in the native AR the efficient recruitment of coactivators requires a functional association of the NTD with the LBD and that the binding of coactivators occurs primarily through the NTD. PMID- 10454557 TI - Activation of silent replication origins at autonomously replicating sequence elements near the HML locus in budding yeast. AB - In the budding yeast, Saccharomyces cerevisiae, replicators can function outside the chromosome as autonomously replicating sequence (ARS) elements; however, within chromosome III, certain ARSs near the transcriptionally silent HML locus show no replication origin activity. Two of these ARSs comprise the transcriptional silencers E (ARS301) and I (ARS302). Another, ARS303, resides between HML and the CHA1 gene, and its function is not known. Here we further localized and characterized ARS303 and in the process discovered a new ARS, ARS320. Both ARS303 and ARS320 are competent as chromosomal replication origins since origin activity was seen when they were inserted at a different position in chromosome III. However, at their native locations, where the two ARSs are in a cluster with ARS302, the I silencer, no replication origin activity was detected regardless of yeast mating type, special growth conditions that induce the transcriptionally repressed CHA1 gene, trans-acting mutations that abrogate transcriptional silencing at HML (sir3, orc5), or cis-acting mutations that delete the E and I silencers containing ARS elements. These results suggest that, for the HML ARS cluster (ARS303, ARS320, and ARS302), inactivity of origins is independent of local transcriptional silencing, even though origins and silencers share key cis- and trans-acting components. Surprisingly, deletion of active replication origins located 25 kb (ORI305) and 59 kb (ORI306) away led to detection of replication origin function at the HML ARS cluster, as well as at ARS301, the E silencer. Thus, replication origin silencing at HML ARSs is mediated by active replication origins residing at long distances from HML in the chromosome. The distal active origins are known to fire early in S phase, and we propose that their inactivation delays replication fork arrival at HML, providing additional time for HML ARSs to fire as origins. PMID- 10454558 TI - The yeast trimeric guanine nucleotide-binding protein alpha subunit, Gpa2p, controls the meiosis-specific kinase Ime2p activity in response to nutrients. AB - Saccharomyces cerevisiae Gpa2p, the alpha subunit of a heterotrimeric guanine nucleotide-binding protein (G protein), is involved in the regulation of vegetative growth and pseudohyphal development. Here we report that Gpa2p also controls sporulation by interacting with the regulatory domain of Ime2p (Sme1p), a protein kinase essential for entrance of meiosis and sporulation. Protein protein interactions between Gpa2p and Ime2p depend on the GTP-bound state of Gpa2p and correlate with down-regulation of Ime2p kinase activity in vitro. Overexpression of Ime2p inhibits pseudohyphal development and enables diploid cells to sporulate even in the presence of glucose or nitrogen. In contrast, overexpression of Gpa2p in cells simultaneously overproducing Ime2p results in a drastic reduction of sporulation efficiency, demonstrating an inhibitory effect of Gpa2p on Ime2p function. Furthermore, deletion of GPA2 accelerates sporulation on low-nitrogen medium. These observations are consistent with the following model. In glucose-containing medium, diploid cells do not sporulate because Ime2p is inactive or expressed at low levels. Upon starvation, expression of Gpa2p and Ime2p is induced but sporulation is prevented as long as nitrogen is present in the medium. The negative control of Ime2p kinase activity is exerted at least in part through the activated form of Gpa2p and is released as soon as nutrients are exhausted. This model attributes a switch function to Gpa2p in the meiosis pseudohyphal growth decision. PMID- 10454559 TI - Regulated expression of focal adhesion kinase-related nonkinase, the autonomously expressed C-terminal domain of focal adhesion kinase. AB - Focal adhesion kinase (FAK) has been implicated in cellular processes that control cell adhesion, migration, cell cycle progression, and apoptosis. FRNK (FAK-related nonkinase) is the autonomously expressed, noncatalytic C-terminal portion of FAK. When ectopically expressed in cells, FRNK has been shown to act as a negative regulator of FAK activity, inhibiting cell spreading, migration, and cell cycle progression. The mechanisms that regulate FRNK expression during embryonic development and the functional role of FRNK in normal cell homeostasis remain poorly understood. Herein we show that FRNK expression in chicken cells is directed by an alternative promoter residing within an intron of FAK, positioned 3' of the exon encoding sequences for the catalytic domain and 5' of the exon encoding sequences for the C-terminal domain of FAK (e.g., FRNK). Using probes specific for FRNK, we show that FRNK expression occurs early in chicken embryogenesis, being readily detected at day 3, 6, or 9. Late in embryogenesis, at day 18, FRNK is expressed in a tissue-specific manner, predominately in lung and intestine cells. Western blot analysis of mouse tissues with a FAK-specific antibody revealed the expression of FRNK in the mouse lung. Reverse transcriptase PCR analysis of mouse lung RNA revealed the presence of spliced FRNK mRNAs containing 5' untranslated sequences derived from a positionally conserved exon present in the mouse genome. FAK is the first example of a tyrosine kinase regulated by a domain under the control of an alternative intronic promoter. It is also the first example of a focal adhesion-associated protein regulated by such a mechanism and thus represents a novel means for the modulation of cell adhesion signaling. PMID- 10454560 TI - Analysis of primary structural determinants that distinguish the centromere specific function of histone variant Cse4p from histone H3. AB - Cse4p is a variant of histone H3 that has an essential role in chromosome segregation and centromere chromatin structure in budding yeast. Cse4p has a unique 135-amino-acid N terminus and a C-terminal histone-fold domain that is more than 60% identical to histone H3 and the mammalian centromere protein CENP A. Cse4p and CENP-A have biochemical properties similar to H3 and probably replace H3 in centromere-specific nucleosomes in yeasts and mammals, respectively. In order to identify regions of Cse4p that distinguish it from H3 and confer centromere function, a systematic site-directed mutational analysis was performed. Nested deletions of the Cse4p N terminus showed that this region of the protein contains at least one essential domain. The C-terminal histone fold domain of Cse4p was analyzed by changing Cse4p amino acids that differ between Cse4p and H3 to the analogous H3 residues. Extensive substitution of contiguous Cse4p residues with H3 counterparts resulted in cell lethality. However, all large lethal substitution alleles could be subdivided into smaller viable alleles, many of which caused elevated rates of mitotic chromosome loss. The results indicate that residues critical for wild-type Cse4p function and high fidelity chromosome transmission are distributed across the entire histone-fold domain. Our findings are discussed in the context of the known structure of H3 within the nucleosome and compared with previous results reported for CENP-A. PMID- 10454561 TI - Identification by in vivo genomic footprinting of a transcriptional switch containing NF-kappaB and Sp1 that regulates the IkappaBalpha promoter. AB - In unstimulated cells, NF-kappaB transcription factors are retained in the cytoplasm by inhibitory IkappaB proteins. Upon stimulation by multiple inducers including cytokines or viruses, IkappaBalpha is rapidly phosphorylated and degraded, resulting in the release of NF-kappaB and the subsequent increase in NF kappaB-regulated gene expression. IkappaBalpha gene expression is also regulated by an NF-kappaB autoregulatory mechanism, via NF-kappaB binding sites in the IkappaBalpha promoter. In previous studies, tetracycline-inducible expression of transdominant repressors of IkappaBalpha (TD-IkappaBalpha) progressively decreased endogenous IkappaBalpha protein levels. In the present study, we demonstrate that expression of TD-IkappaBalpha blocked phorbol myristate acetate phytohemagglutinin or tumor necrosis factor alpha-induced IkappaBalpha gene transcription and abolished NF-kappaB DNA binding activity, due to the continued cytoplasmic sequestration of RelA(p65) by TD-IkappaBalpha. In vivo genomic footprinting revealed stimulus-responsive protein-DNA binding not only to the -63 to -53 kappaB1 site but also to the adjacent -44 to -36 Sp1 site of the IkappaBalpha promoter. In vivo protection of both sites was inhibited by tetracycline-inducible TD-IkappaBalpha expression. Prolonged NF-kappaB binding and a temporal switch in the composition of NF-kappaB complexes bound to the -63 to -53 kappaB1 site of the IkappaBalpha promoter were also observed; with time after induction, decreased levels of transcriptionally active p50-p65 and increased p50-c-Rel heterodimers were detected at the kappaB1 site. Mutation of either the kappaB1 site or the Sp1 site abolished transcription factor binding to the respective sites and the inducibility of the IkappaBalpha promoter in transient transfection studies. These observations provide the first in vivo characterization of a promoter proximal transcriptional switch involving NF kappaB and Sp1 that is essential for autoregulation of the IkappaBalpha promoter. PMID- 10454562 TI - MCM proteins are associated with RNA polymerase II holoenzyme. AB - MCMs are a family of proteins related to ATP-dependent helicases that bind to origin recognition complexes and are required for initiation of DNA replication. We report that antibodies against MCM2(BM28) specifically inhibited transcription by RNA polymerase II (Pol II) in microinjected Xenopus oocytes. Consistent with this observation, MCM2 and other MCMs copurified with Pol II and general transcription factors (GTFs) in high-molecular-weight holoenzyme complexes isolated from Xenopus oocytes and HeLa cells. Pol II and GTFs also copurified with MCMs isolated by anti-MCM3 immunoaffinity chromatography. MCMs were specifically displaced from the holoenzyme complex by antibody against the C terminal domain (CTD) of Pol II. In addition, MCMs bound to a CTD affinity column, suggesting that their association with holoenzyme depends in part on this domain of Pol II. These results suggest a new function for MCM proteins as components of the Pol II transcriptional apparatus. PMID- 10454563 TI - Multiple signal input and output domains of the 160-kilodalton nuclear receptor coactivator proteins. AB - Members of the 160-kDa nuclear receptor coactivator family (p160 coactivators) bind to the conserved AF-2 activation function found in the hormone binding domains of nuclear receptors (NR) and are potent transcriptional coactivators for NRs. Here we report that the C-terminal region of p160 coactivators glucocorticoid receptor interacting protein 1 (GRIP1), steroid receptor coactivator 1 (SRC-1a), and SRC-1e binds the N-terminal AF-1 activation function of the androgen receptor (AR), and p160 coactivators can thereby enhance transcriptional activation by AR. While they all interact efficiently with AR AF 1, these same coactivators have vastly different binding strengths with and coactivator effects on AR AF-2. p160 activation domain AD1, which binds secondary coactivators CREB binding protein (CBP) and p300, was previously implicated as the principal domain for transmitting the activating signal to the transcription machinery. We identified a new highly conserved motif in the AD1 region which is important for CBP/p300 binding. Deletion of AD1 only partially reduced p160 coactivator function, due to signaling through AD2, another activation domain located at the C-terminal end of p160 coactivators. C-terminal coactivator fragments lacking AD1 but containing AD2 and the AR AF-1 binding site served as efficient coactivators for full-length AR and AR AF-1. The two signal input domains (one that binds NR AF-2 domains and one that binds AF-1 domains of some but not all NRs) and the two signal output domains (AD1 and AD2) of p160 coactivators played different relative roles for two different NRs: AR and thyroid hormone receptor. PMID- 10454564 TI - Identification of cis and trans elements involved in the cell cycle regulation of multiple genes in Crithidia fasciculata. AB - Transcripts of several DNA replication genes, including the RPA1 and TOP2 genes, encoding the large subunit of nuclear replication protein A and the kinetoplast topoisomerase II, accumulate periodically during the cell cycle in the trypanosomatid Crithidia fasciculata. An octamer consensus sequence, CAUAGAAG, present in the 5' untranslated regions (UTR) of these mRNAs is required for periodic accumulation of the TOP2 and RPA1 transcripts and also for binding of a nuclear factor(s) to the 5' UTR RNAs of these genes. We show here that insertion of multiple (six) copies of this octamer sequence (6x octamer) into the 5' UTR of a reporter gene confers periodic accumulation on its transcript. Competition experiments and UV cross-linking studies show that the 6x octamer RNA and TOP2 5' UTR RNA bind to the same nuclear factor(s). Single-nucleotide substitutions in the 6x octamer that abolish the RNA gel shift also prevent cyclic accumulation of the reporter gene transcript. A protein termed cycling element binding protein, purified by affinity chromatography using 6x octamer RNA as a ligand, binds to RNAs containing wild-type octamers and not to those with mutant octamers. These results define a small sequence element in C. fasciculata mRNAs required for their cell cycle regulation and report the identification and purification of a putative regulatory protein that binds specifically to these elements. PMID- 10454565 TI - Ectopic expression of Cdc25A accelerates the G(1)/S transition and leads to premature activation of cyclin E- and cyclin A-dependent kinases. AB - Human Cdc25 phosphatases play important roles in cell cycle regulation by removing inhibitory phosphates from tyrosine and threonine residues of cyclin dependent kinases. Three human Cdc25 isoforms, A, B, and C, have been discovered. Cdc25B and Cdc25C play crucial roles at the G(2)/M transition. In the present study, we have investigated the function of human Cdc25A phosphatase. Cell lines that express human Cdc25A in an inducible manner have been generated. Ectopic expression of Cdc25A accelerates the G(1)/S-phase transition, indicating that Cdc25A controls an event(s) that is rate limiting for entry into S phase. Furthermore, we carried out a detailed analysis of the expression and activation of human Cdc25A. Activation of endogenous Cdc25A occurs during late G(1) phase and increases in S and G(2) phases. We further demonstrate that Cdc25A is activated at the same time as cyclin E- and cyclin A-dependent kinases. In vitro, Cdc25A dephosphorylates and activates the cyclin-Cdk complexes that are active during G(1). Overexpression of Cdc25A in the inducible system, however, leads to a premature activation of both cyclin E-Cdk2 and cyclin A-Cdk2 complexes, while no effect of cyclin D-dependent kinases is observed. Furthermore, Cdc25A overexpression induces a tyrosine dephosphorylation of Cdk2. These results suggest that Cdc25A is an important regulator of the G(1)/S-phase transition and that cyclin E- and cyclin A-dependent kinases act as direct targets. PMID- 10454566 TI - The antiapoptotic gene mcl-1 is up-regulated by the phosphatidylinositol 3 kinase/Akt signaling pathway through a transcription factor complex containing CREB. AB - mcl-1 is an immediate-early gene activated by the granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 3 (IL-3) signaling pathways and plays an important role in the viability response of these cytokines. In this study, we demonstrated that cytokine stimulation of mcl-1 mRNA and protein expression were attenuated by pretreatment of cells with phosphatidylinositol 3-kinase (PI3-K) inhibitors. Reporter gene assays further showed that the PI3-K/Akt signaling pathway was involved in IL-3 activation of mcl-1 gene transcription. Analysis of the mcl-1 promoter revealed that both promoter elements, SIE at position -87 and CRE-2 at -70, contribute to IL-3 stimulation of mcl-1 gene expression. Although either the SIE site or the CRE-2 site alone was sufficient to confer IL-3 inducibility on a heterologous promoter, only IL-3 activation of the CRE-2 reporter was mediated via the PI3-K/Akt pathway. The SIE binding activity was constitutively high in cells deprived of or stimulated by IL-3. In contrast, the CRE-2 binding activity was low in cytokine-starved cells and was strongly induced within 1 h following cytokine treatment of cells. In addition, cytokine induction of the CRE-2 but not of the SIE binding activity was dependent on activation of the PI3-K/Akt signaling pathway. Lastly, we showed that CREB was one component of the CRE-2 binding complex and played a role in IL-3 activation of the mcl-1 reporter gene. Taken together, our results suggest that both PI3-K/Akt-dependent and -independent pathways contribute to the IL-3 activation of mcl-1 gene expression. Activation of mcl-1 by the PI3-K/Akt-dependent pathway is through a transcription factor complex containing CREB. PMID- 10454567 TI - Two inactive fragments of the integral RNA cooperate to assemble active telomerase with the human protein catalytic subunit (hTERT) in vitro. AB - We have mapped the 5' and 3' boundaries of the region of the human telomerase RNA (hTR) that is required to produce activity with the human protein catalytic subunit (hTERT) by using in vitro assembly systems derived from rabbit reticulocyte lysates and human cell extracts. The region spanning nucleotides +33 to +325 of the 451-base hTR is the minimal sequence required to produce levels of telomerase activity that are comparable with that made with full-length hTR. Our results suggest that the sequence approximately 270 bases downstream of the template is required for efficient assembly of active telomerase in vitro; this sequence encompasses a substantially larger portion of the 3' end of hTR than previously thought necessary. In addition, we identified two fragments of hTR (nucleotides +33 to +147 and +164 to +325) that cannot produce telomerase activity when combined separately with hTERT but can function together to assemble active telomerase. These results suggest that the minimal sequence of hTR can be divided into two sections, both of which are required for de novo assembly of active telomerase in vitro. PMID- 10454568 TI - Grb10, a positive, stimulatory signaling adapter in platelet-derived growth factor BB-, insulin-like growth factor I-, and insulin-mediated mitogenesis. AB - Grb10 has been described as a cellular partner of several receptor tyrosine kinases, including the insulin receptor (IR) and the insulin-like growth factor I (IGF-I) receptor (IGF-IR). Its cellular role is still unclear and a positive as well as an inhibitory role in mitogenesis depending on the cell context has been implicated. We have tested other mitogenic receptor tyrosine kinases as putative Grb10 partners and have identified the activated forms of platelet-derived growth factor (PDGF) receptor beta (PDGFRbeta), hepatocyte growth factor receptor (Met), and fibroblast growth factor receptor as candidates. We have mapped Y771 as a PDFGRbeta site that is involved in the association with Grb10 via its SH2 domain. We have further investigated the putative role of Grb10 in mitogenesis with four independent experimental strategies and found that all consistently suggested a role as a positive, stimulatory signaling adaptor in normal fibroblasts. (i) Complete Grb10 expression from cDNA with an ecdysone-regulated transient expression system stimulated PDGF-BB-, IGF-I, and insulin- but not epidermal growth factor (EGF)-induced DNA synthesis in an ecdysone dose-responsive fashion. (ii) Microinjection of the (dominant-negative) Grb10 SH2 domain interfered with PDGF-BB- and insulin-induced DNA synthesis. (iii) Alternative experiments were based on cell-permeable fusion peptides with the Drosophila antennapedia homeodomain which effectively traverse the plasma membrane of cultured cells. A cell-permeable Grb10 SH2 domain similarly interfered with PDGF-BB-, IGF-I-, and insulin-induced DNA synthesis. In contrast, a cell-permeable Grb10 Pro-rich putative SH3 domain binding region interfered with IGF-I- and insulin- but not with PDGF-BB- or EGF-induced DNA synthesis. (iv) Transient overexpression of complete Grb10 increased whereas cell-permeable Grb10 SH2 domain fusion peptides substantially decreased the cell proliferation rate (as measured by cell numbers) in normal fibroblasts. These experimental strategies independently suggest that Grb10 functions as a positive, stimulatory, mitogenic signaling adapter in PDGF BB, IGF-I, and insulin action. This function appears to involve the Grb10 SH2 domain, a novel sequence termed BPS, and the Pro-rich putative SH3 domain binding region in IGF-I- and insulin-mediated mitogenesis. In contrast, PDGF-BB-mediated mitogenesis appears to depend on the SH2 but not on the Pro-rich region and may involve other, unidentified Grb10 domains. Distinct protein domains may help to define specific Grb10 functions in different signaling pathways. PMID- 10454569 TI - Elevated cyclin E levels, inactive retinoblastoma protein, and suppression of the p27(KIP1) inhibitor characterize early development of promyeloid cells into macrophages. AB - Cyclin-dependent kinase inhibitors such as p27(KIP1) have recently been shown to lead to cellular differentiation by causing cell cycle arrest, but it is unknown whether similar events occur in differentiating promyeloid cells. Hematopoietic progenitor cells undergo lineage-restricted differentiation, which is accompanied by expression of distinct maturation markers. Here we show that the classical growth factor insulin-like growth factor I (IGF-I) potently promotes vitamin D(3) induced macrophage differentiation of promyeloid cells, as assessed by measurement of a coordinate increase in expression of the integrin alpha subunit CD11b, the CD14 lipopolysaccharide receptor, and the macrophage-specific esterase, alpha-naphthyl acetate esterase, as early as 24 h following initiation of terminal differentiation. Addition of IGF-I to cells undergoing vitamin D(3) induced differentiation also leads to an early increase in expression of cyclin E, phosphorylation of the retinoblastoma tumor suppressor protein, and a doubling of the cell number. Early expression of CD11b (24 h) is simultaneously accompanied by inhibition in the expression of p27(KIP1). Cell cycle analysis with propidium iodide revealed that CD11b expression at 24 h following initiation of differentiation occurs at all phases of the cell cycle instead of only those cells arrested in G(0)/G(1). Similarly, development of a novel double-labeling intra- and extracellular flow-cytometric technique demonstrated that single cells expressing the mature leukocyte differentiation antigen CD11b can also incorporate the thymidine analog bromodeoxyuridine. Likewise, expression of the intracellular DNA polymerase delta cofactor/proliferating-cell nuclear antigen at 24 h is also simultaneously expressed with the surface marker CD11b, indicating that these cells continue to proliferate early in their differentiation program. Finally, at 24 h following induction of differentiation, IGF-I promoted a fourfold increase in the uptake of [(3)H]thymidine by purified populations of CD11b-expressing cells. Taken together, these data demonstrate that the initial steps associated with terminal macrophage differentiation occur concomitantly with progression through the cell cycle and that these very early differentiation events do not require the accumulation of p27(KIP1). PMID- 10454571 TI - In vitro import of a nuclearly encoded tRNA into the mitochondrion of Trypanosoma brucei. AB - All of the mitochondrial tRNAs of Trypanosoma brucei have been shown to be encoded in the nucleus and must be imported into the mitochondrion. The import of nuclearly encoded tRNAs into the mitochondrion has been demonstrated in a variety of organisms and is essential for proper function in the mitochondrion. An in vitro import assay has been developed to study the pathway of tRNA import in T. brucei. The in vitro system utilizes crude isolated trypanosome mitochondria and synthetic RNAs transcribed from a cloned nucleus-encoded tRNA gene cluster. The substrate, composed of tRNA(Ser) and tRNA(Leu), is transcribed in tandem with a 59-nucleotide intergenic region. The tandem tRNA substrate is imported rapidly, while the mature-size tRNA(Leu) fails to be imported in this system. These results suggest that the preferred substrate for tRNA import into trypanosome mitochondria is a precursor molecule composed of tandemly linked tRNAs. Import of the tandem tRNA substrate requires (i) a protein component that is associated with the surface of the mitochondrion, (ii) ATP pools both outside and within the mitochondrion, and (iii) a membrane potential. Dissipation of the proton gradient across the inner mitochondrial membrane by treatment with an uncoupling agent inhibits import of the tandem tRNA substrate. Characterization of the import requirements indicates that mitochondrial RNA import proceeds by a pathway including a protein component associated with the outer mitochondrial membrane, ATP-dependent steps, and a mitochondrial membrane potential. PMID- 10454570 TI - Role of distinct mitogen-activated protein kinase pathways and cooperation between Ets-2, ATF-2, and Jun family members in human urokinase-type plasminogen activator gene induction by interleukin-1 and tetradecanoyl phorbol acetate. AB - We have investigated the in vivo and in vitro regulation of the human urokinase type plasminogen activator (uPA) gene by interleukin-1 (IL-1) and analyzed the transcription factors and signalling pathways involved in the response of the 2.0-kb uPA enhancer to IL-1 induction and to tetradecanoyl phorbol acetate (TPA) induction. Mutational analysis showed the cooperative activity of the Ets-binding site (EBS) and the two AP-1 elements of the enhancer. The results reveal that the EBS is required for the response to both inducers mediated by Ets-2, which is regulated at a level subsequent to DNA binding, by an IL-1- and phorbol ester inducible transactivation domain. Both the IL-1 and the TPA-mediated induction result in a drastic increase of AP-1 binding to the downstream site of the enhancer (uPA 3' TPA-responsive element), while a mostly qualitative change, resulting from the interplay between ATF-2 homodimers and c-Jun-ATF-2 heterodimers, takes place at the upstream AP-1 element. The analysis of two distinct mitogen-activated protein kinase pathways shows that stress-activated protein kinase-Jun N-terminal kinase activation, resulting in the phosphorylation of ATF-2, c-Jun, and JunD, is required not only for the IL-1- but also for the TPA-dependent induction, while the extracellular signal-related kinase 1 (ERK-1) and ERK-2 activation is involved in the TPA- but not in the IL-1-dependent stimulation of the uPA enhancer. PMID- 10454572 TI - Involvement of DNA end-binding protein Ku in Ty element retrotransposition. AB - Saccharomyces cerevisiae Ty elements are retrotransposons whose life cycles are strikingly similar to those of retroviruses. They transpose via an RNA intermediate that is converted to linear double-stranded cDNA and then inserted into the host genome. Although Ty integration is mediated by the element-encoded integrase, it has been proposed that host factors are involved in this process. Here, we show that the DNA end-binding protein Ku, which functions in DNA double strand break repair, potentiates retrotransposition. Specifically, by using a galactose-inducible Ty1 system, we found that in vivo, Ty1 retrotransposition rates were substantially reduced in the absence of Ku. In contrast, this phenotype was not observed with yeast strains containing mutations in other genes that are involved in DNA repair. We present evidence that Ku associates with Ty1 viruslike particles both in vitro and in vivo. These results provide an additional role for Ku and suggest that it might function in the life cycles of retroelements in other systems. PMID- 10454573 TI - The Drosophila melanogaster DmRAD54 gene plays a crucial role in double-strand break repair after P-element excision and acts synergistically with Ku70 in the repair of X-ray damage. AB - The RAD54 gene has an essential role in the repair of double-strand breaks (DSBs) via homologous recombination in yeast as well as in higher eukaryotes. A Drosophila melanogaster strain deficient in the RAD54 homolog DmRAD54 is characterized by increased X-ray and methyl methanesulfonate (MMS) sensitivity. In addition, DmRAD54 is involved in the repair of DNA interstrand cross-links, as is shown here. However, whereas X-ray-induced loss-of-heterozygosity (LOH) events were completely absent in DmRAD54(-/-) flies, treatment with cross-linking agents or MMS resulted in only a slight reduction in LOH events in comparison with those in wild-type flies. To investigate the relative contributions of recombinational repair and nonhomologous end joining in DSB repair, a DmRad54(-/-)/DmKu70(-/-) double mutant was generated. Compared with both single mutants, a strong synergistic increase in X-ray sensitivity was observed in the double mutant. No similar increase in sensitivity was seen after treatment with MMS. Apparently, the two DSB repair pathways overlap much less in the repair of MMS-induced lesions than in that of X-ray-induced lesions. Excision of P transposable elements in Drosophila involves the formation of site-specific DSBs. In the absence of the DmRAD54 gene product, no male flies could be recovered after the excision of a single P element and the survival of females was reduced to 10% compared to that of wild-type flies. P-element excision involves the formation of two DSBs which have identical 3' overhangs of 17 nucleotides. The crucial role of homologous recombination in the repair of these DSBs may be related to the very specific nature of the breaks. PMID- 10454575 TI - Insulin-induced phosphorylation and activation of cyclic nucleotide phosphodiesterase 3B by the serine-threonine kinase Akt. AB - Cyclic nucleotide phosphodiesterase (PDE) is an important regulator of the cellular concentrations of the second messengers cyclic AMP (cAMP) and cGMP. Insulin activates the 3B isoform of PDE in adipocytes in a phosphoinositide 3 kinase-dependent manner; however, downstream effectors that mediate signaling to PDE3B remain unknown. Insulin-induced phosphorylation and activation of endogenous or recombinant PDE3B in 3T3-L1 adipocytes have now been shown to be inhibited by a dominant-negative mutant of the serine-threonine kinase Akt, suggesting that Akt is necessary for insulin-induced phosphorylation and activation of PDE3B. Serine-273 of mouse PDE3B is located within a motif (RXRXXS) that is preferentially phosphorylated by Akt. A mutant PDE3B in which serine-273 was replaced by alanine was not phosphorylated either in response to insulin in intact cells or by purified Akt in vitro. In contrast, PDE3B mutants in which alanine was substituted for either serine-296 or serine-421, each of which lies within a sequence (RRXS) preferentially phosphorylated by cAMP-dependent protein kinase, were phosphorylated by Akt in vitro or in response to insulin in intact cells. Moreover, the serine-273 mutant of PDE3B was not activated by insulin when expressed in adipocytes. These results suggest that PDE3B is a physiological substrate of Akt and that Akt-mediated phosphorylation of PDE3B on serine-273 is important for insulin-induced activation of PDE3B. PMID- 10454574 TI - RanGTP-regulated interactions of CRM1 with nucleoporins and a shuttling DEAD-box helicase. AB - CRM1 is an export receptor mediating rapid nuclear exit of proteins and RNAs to the cytoplasm. CRM1 export cargoes include proteins with a leucine-rich nuclear export signal (NES) that bind directly to CRM1 in a trimeric complex with RanGTP. Using a quantitative CRM1-NES cargo binding assay, significant differences in affinity for CRM1 among natural NESs are demonstrated, suggesting that the steady state nucleocytoplasmic distribution of shuttling proteins could be determined by the relative strengths of their NESs. We also show that a trimeric CRM1-NES RanGTP complex is disassembled by RanBP1 in the presence of RanGAP, even though RanBP1 itself contains a leucine-rich NES. Selection of CRM1-binding proteins from Xenopus egg extract leads to the identification of an NES-containing DEAD box helicase, An3, that continuously shuttles between the nucleus and the cytoplasm. In addition, we identify the Xenopus homologue of the nucleoporin CAN/Nup214 as a RanGTP- and NES cargo-specific binding site for CRM1, suggesting that this nucleoporin plays a role in export complex disassembly and/or CRM1 recycling. PMID- 10454576 TI - The Ras mutant D119N is both dominant negative and activated. AB - The introduction of mutation D119N (or its homolog) in the NKxD nucleotide binding motif of various Ras-like proteins produces constitutively activated or dominant-negative effects, depending on the system and assay. Here we show that Ras(D119N) has an inhibitory effect at a cell-specific concentration in PC12 and NIH 3T3 cells. Biochemical data strongly suggest that the predominant effect of mutation D119N in Ras-a strong decrease in nucleotide affinity-enables this mutant (i) to sequester its guanine nucleotide exchange factor, as well as (ii) to rapidly bind GTP, independent of the regulatory action of the exchange factor. Since mutation D119N does not affect the interaction between Ras and effector molecules, the latter effect causes Ras(D119N) to act as an activated Ras protein at concentrations higher than that of the exchange factor. In comparison, Ras(S17N), which also shows a strongly decreased nucleotide affinity, does not bind to effector molecules. These results point to two important prerequisites of dominant-negative Ras mutants: an increased relative affinity of the mutated Ras for the exchange factor over that for the nucleotide and an inability to interact with the effector or effectors. Remarkably, the introduction of a second, partial loss-of-function, mutation turns Ras(D119N) into a strong dominant-negative mutant even at high concentrations, as demonstrated by the inhibitory effects of Ras(E37G/D119N) on nerve growth factor-mediated neurite outgrowth in PC12 cells and Ras(T35S/D119N) on fetal calf serum-mediated DNA synthesis in NIH 3T3 cells. Interpretations of these results are discussed. PMID- 10454577 TI - Identification of novel import and export signals of human TAP, the protein that binds to the constitutive transport element of the type D retrovirus mRNAs. AB - The nuclear export of the unspliced type D retrovirus mRNA depends on the cis acting constitutive transport RNA element (CTE) that has been shown to interact with the human TAP (hTAP) protein promoting the export of the CTE-containing mRNAs. We report here that hTAP is a 619-amino-acid protein extending the previously identified protein by another 60 residues at the N terminus and that hTAP shares high homology with the predicted rat and mouse TAP proteins. We found that hTAP is a nuclear protein that accumulates in the nuclear rim and the nucleoplasm. We further demonstrated that hTAP is able to shuttle between the nucleus and the cytoplasm. Identification of the signals responsible for nuclear import (NLS) and export (NES) revealed that they are distinct but partially overlapping. NLS and NES of hTAP are active transferable signals that do not share similarities with known elements. The C-terminal portion contributes further to hTAP's nuclear retention and contains a signal(s) for nuclear rim association. Taken together, our data show that hTAP is a dynamic protein capable of bidirectional trafficking across the nuclear envelope. These data further support hTAP's role as an export factor of the CTE-containing mRNAs. PMID- 10454578 TI - Phenobarbital-responsive nuclear translocation of the receptor CAR in induction of the CYP2B gene. AB - The constitutively active receptor (CAR) transactivates a distal enhancer called the phenobarbital (PB)-responsive enhancer module (PBREM) found in PB-inducible CYP2B genes. CAR dramatically increases its binding to PBREM in livers of PB treated mice. We have investigated the cellular mechanism of PB-induced increase of CAR binding. Western blot analyses of mouse livers revealed an extensive nuclear accumulation of CAR following PB treatment. Nuclear contents of CAR perfectly correlate with an increase of CAR binding to PBREM. PB-elicited nuclear accumulation of CAR appears to be a general step regulating the induction of CYP2B genes, since treatments with other PB-type inducers result in the same nuclear accumulation of CAR. Both immunoprecipitation and immunohistochemistry studies show cytoplasmic localization of CAR in the livers of nontreated mice, indicating that CAR translocates into nuclei following PB treatment. Nuclear translocation of CAR also occurs in mouse primary hepatocytes but not in hepatocytes treated with the protein phosphatase inhibitor okadaic acid. Thus, the CAR-mediated transactivation of PBREM in vivo becomes PB responsive through an okadaic acid-sensitive nuclear translocation process. PMID- 10454579 TI - Activating signal cointegrator 1, a novel transcription coactivator of nuclear receptors, and its cytosolic localization under conditions of serum deprivation. AB - Activating signal cointegrator 1 (ASC-1) harbors an autonomous transactivation domain that contains a putative zinc finger motif which provides binding sites for basal transcription factors TBP and TFIIA, transcription integrators steroid receptor coactivator 1 (SRC-1) and CBP-p300, and nuclear receptors, as demonstrated by the glutathione S-transferase pull-down assays and the yeast two hybrid tests. The ASC-1 binding sites involve the hinge domain but not the C terminal AF2 core domain of nuclear receptors. Nonetheless, ASC-1 appears to require the AF2-dependent factors to function (i.e., CBP-p300 and SRC-1), as suggested by the ability of ASC-1 to coactivate nuclear receptors, either alone or in cooperation with SRC-1 and p300, as well as its inability to coactivate a mutant receptor lacking the AF2 core domain. By using indirect immunofluorescence, we further show that ASC-1, a nuclear protein, is localized to the cytoplasm under conditions of serum deprivation but is retained in the nucleus when it is serum starved in the presence of ligand or coexpressed CBP or SRC-1. These results suggest that ASC-1 is a novel coactivator molecule of nuclear receptors which functions in conjunction with CBP-p300 and SRC-1 and may play an important role in establishing distinct coactivator complexes under different cellular conditions. PMID- 10454580 TI - Differential roles of Akt, Rac, and Ral in R-Ras-mediated cellular transformation, adhesion, and survival. AB - Multiple biological functions have been ascribed to the Ras-related G protein R Ras. These include the ability to transform NIH 3T3 fibroblasts, the promotion of cell adhesion, and the regulation of apoptotic responses in hematopoietic cells. To investigate the signaling mechanisms responsible for these biological phenotypes, we compared three R-Ras effector loop mutants (S61, G63, and C66) for their relative biological and biochemical properties. While the S61 mutant retained the ability to cause transformation, both the G63 and the C66 mutants were defective in this biological activity. On the other hand, while both the S61 and the C66 mutants failed to promote cell adhesion and survival in 32D cells, the G63 mutant retained the ability to induce these biological activities. Thus, the ability of R-Ras to transform cells could be dissociated from its propensity to promote cell adhesion and survival. Although the transformation-competent S61 mutant bound preferentially to c-Raf, it only weakly stimulated the mitogen activated protein kinase (MAPK) activity, and a dominant negative mutant of MEK did not significantly perturb R-Ras oncogenicity. Instead, a dominant negative mutant of phosphatidylinositol 3-kinase (PI3-K) drastically inhibited the oncogenic potential of R-Ras. Interestingly, the ability of the G63 mutant to induce cell adhesion and survival was closely associated with the PI3-K-dependent signaling cascades. To further delineate R-Ras downstream signaling events, we observed that while a dominant negative mutant of Akt/protein kinase inhibited the ability of R-Ras to promote cell survival, both dominant negative mutants of Rac and Ral suppressed cell adhesion stimulated by R-Ras. Thus, the biological actions of R-Ras are mediated by multiple effectors, with PI3-K-dependent signaling cascades being critical to its functions. PMID- 10454582 TI - The oncoprotein E2A-Pbx1a collaborates with Hoxa9 to acutely transform primary bone marrow cells. AB - A recurrent translocation between chromosome 1 (Pbx1) and 19 (E2A) leading to the expression of the E2A-Pbx1 fusion oncoprotein occurs in approximately 5 to 10% of acute leukemias in humans. It has been proposed that some of the oncogenic potential of E2A-Pbx1 could be mediated through heterocomplex formation with Hox proteins, which are also involved in human and mouse leukemias. To directly test this possibility, mouse bone marrow cells were engineered by retroviral gene transfer to overexpress E2A-Pbx1a together with Hoxa9. The results obtained demonstrated a strong synergistic interaction between E2A-Pbx1a and Hoxa9 in inducing growth factor-independent proliferation of transduced bone marrow cells in vitro and leukemic growth in vivo in only 39 +/- 2 days. The leukemic blasts which coexpress E2A-Pbx1a and Hoxa9 showed little differentiation and produced cytokines such as interleukin-3, granulocyte colony-stimulating factor, and Steel. Together, these studies demonstrate that the Hoxa9 and E2A-Pbx1a gene products collaborate to produce a highly aggressive acute leukemic disease. PMID- 10454581 TI - Direct association and nuclear import of the hepatitis B virus X protein with the NF-kappaB inhibitor IkappaBalpha. AB - The X protein of hepatitis B virus (HBV) is a transcriptional activator which is required for infection and may play an important role in HBV-associated hepatocarcinogenesis. It has been suggested that X acts as a nuclear coactivator or stimulates several signal transduction pathways by acting in the cytoplasm. One of these pathways leads to the nuclear translocation of NF-kappaB. A recent report indicates that X activates NF-kappaB by acting on two cytoplasmic inhibitors of this family of transcription factors: IkappaBalpha and the precursor/inhibitor p105. We demonstrate here that X directly interacts with IkappaBalpha, which is able to transport it to the nucleus by a piggyback mechanism. This transport requires a region of IkappaBalpha (the second ankyrin repeat) which has been demonstrated to be involved in its nuclear import following NF-kappaB activation. Using deletion mutants, we showed that amino acids 249 to 253 of IkappaBalpha (located in the C-terminal part of the sixth ankyrin repeat) play a critical role in the interaction with X. This small region overlaps one of the domains of IkappaBalpha mediating the interaction with the p50 and p65 subunits of NF-kappaB and is also close to the nuclear export sequence of IkappaBalpha, therefore providing a potential explanation for the nuclear accumulation of IkappaBalpha with X. This association can also be observed upon the induction of endogenous IkappaBalpha by tumor necrosis factor alpha (TNF-alpha) treatment of Chang cells expressing X. In accordance with this observation, band shift analysis indicates that X induces a sustained NF-kappaB activation following TNF-alpha treatment, probably by preventing the reassociation of newly synthesized nuclear IkappaBalpha with DNA-bound NF-kappaB complexes. PMID- 10454583 TI - Transcriptional activation by NF-kappaB requires multiple coactivators. AB - Nuclear factor-kappaB (NF-kappaB) plays a role in the transcriptional regulation of genes involved in inflammation and cell survival. In this report we demonstrate that NF-kappaB recruits a coactivator complex that has striking similarities to that recruited by nuclear receptors. Inactivation of either cyclic AMP response element binding protein (CREB)-binding protein (CBP), members of the p160 family of coactivators, or the CBP-associated factor (p/CAF) by nuclear antibody microinjection prevents NF-kappaB-dependent transactivation. Like nuclear receptor-dependent gene expression, NF-kappaB-dependent gene expression requires specific LXXLL motifs in one of the p160 family members, and enhancement of NF-kappaB activity requires the histone acetyltransferase (HAT) activity of p/CAF but not that of CBP. This coactivator complex is differentially recruited by members of the Rel family. The p50 homodimer fails to recruit coactivators, although the p50-p65 heterodimeric form of the transcription factor assembles the integrator complex. These findings provide new mechanistic insights into how this family of dimeric transcription factors has a differential effect on gene expression. PMID- 10454584 TI - CDC25A phosphatase is a target of E2F and is required for efficient E2F-induced S phase. AB - Functional inactivation of the pRB pathway is a very frequent event in human cancer, resulting in deregulated activity of the E2F transcription factors. To understand the functional role of the E2Fs in cell proliferation, we have developed cell lines expressing E2F-1, E2F-2, and E2F-3 fused to the estrogen receptor ligand binding domain (ER). In this study, we demonstrated that activation of all three E2Fs could relieve the mitogen requirement for entry into S phase in Rat1 fibroblasts and that E2F activity leads to a shortening of the G(0)-G(1) phase of the cell cycle by 6 to 7 h. In contrast to the current assumption that E2F-1 is the only E2F capable of inducing apoptosis, we showed that deregulated E2F-2 and E2F-3 activities also result in apoptosis. Using the ERE2F-expressing cell lines, we demonstrated that several genes containing E2F DNA binding sites are efficiently induced by the E2Fs in the absence of protein synthesis. Furthermore, CDC25A is defined as a novel E2F target whose expression can be directly regulated by E2F-1. Data showing that CDC25A is an essential target for E2F-1, since its activity is required for efficient induction of S phase by E2F-1, are provided. Finally, our results show that expression of two E2F target genes, namely CDC25A and cyclin E, is sufficient to induce entry into S phase in quiescent fibroblasts. Taken together, our results provide an important step in defining how E2F activity leads to deregulated proliferation. PMID- 10454585 TI - Suppression of STAT5 functions in liver, mammary glands, and T cells in cytokine inducible SH2-containing protein 1 transgenic mice. AB - Various cytokines utilize Janus kinase (JAK) and the STAT (signal transducers and activators of transcription) family of transcription factors to carry out their biological functions. Among STATs, two highly related proteins, STAT5a and STAT5b, are activated by various cytokines, including prolactin, growth hormone, erythropoietin, interleukin 2 (IL-2), and IL-3. We have cloned a STAT5-dependent immediate-early cytokine-responsive gene, CIS1 (encoding cytokine-inducible SH2 containing protein 1). In this study, we created CIS1 transgenic mice under the control of a beta-actin promoter. The transgenic mice developed normally; however, their body weight was lower than that of the wild-type mice, suggesting a defect in growth hormone signaling. Female transgenic mice failed to lactate after parturition because of a failure in terminal differentiation of the mammary glands, suggesting a defect in prolactin signaling. The IL-2-dependent upregulation of the IL-2 receptor alpha chain and proliferation were partially suppressed in the T cells of transgenic mice. These phenotypes remarkably resembled those found in STAT5a and/or STAT5b knockout mice. Indeed, STAT5 tyrosine phosphorylation was suppressed in mammary glands and the liver. Furthermore, the IL-2-induced activation of STAT5 was markedly inhibited in T cells in transgenic mice, while leukemia inhibitory factor-induced STAT3 phosphorylation was not affected. We also found that the numbers of gamma delta T cells, as well as those of natural killer (NK) cells and NKT cells, were dramatically decreased and that Th1/Th2 differentiation was altered in transgenic mice. These data suggest that CIS1 functions as a specific negative regulator of STAT5 in vivo and plays an important regulatory role in the liver, mammary glands, and T cells. PMID- 10454586 TI - E2F1 has both oncogenic and tumor-suppressive properties in a transgenic model. AB - Using a transgenic mouse model expressing the E2F1 gene under the control of a keratin 5 (K5) promoter, we previously demonstrated that increased E2F1 activity can promote tumorigenesis by cooperating with either a v-Ha-ras transgene to induce benign skin papillomas or p53 deficiency to induce spontaneous skin carcinomas. We now report that as K5 E2F1 transgenic mice age, they are predisposed to develop spontaneous tumors in a variety of K5-expressing tissues, including the skin, vagina, forestomach, and odontogenic epithelium. On the other hand, K5 E2F1 transgenic mice are found to be resistant to skin tumor development following a two-stage carcinogenesis protocol. Additional experiments suggest that this tumor-suppressive effect of E2F1 occurs at the promotion stage and may involve the induction of apoptosis. These findings demonstrate that increased E2F1 activity can either promote or inhibit tumorigenesis, dependent upon the experimental context. PMID- 10454587 TI - Methylation-mediated transcriptional silencing in euchromatin by methyl-CpG binding protein MBD1 isoforms. AB - DNA methylation of promoter-associated CpG islands is involved in the transcriptional repression of vertebrate genes. To investigate the mechanisms underlying gene inactivation by DNA methylation, we characterized a human MBD1 protein, one of the components of MeCP1, which possesses a methyl-CpG binding domain (MBD) and cysteine-rich (CXXC) domains. Four novel MBD1 isoforms (MBD1v1, MBD1v2, MBD1v3, and MBD1v4) were identified by the reverse transcription-PCR method. We found that these transcripts were alternatively spliced in the region of CXXC domains and the C terminus. Green fluorescent protein-fused MBD1 was localized to multiple foci on the human genome, mostly in the euchromatin regions, and particularly concentrated in the pericentromeric region of chromosome 1. Both the MBD sequence and genome methylation were required for proper localization of the MBD1 protein. We further investigated whether MBD1 isoforms are responsible for transcriptional repression of human genes. A bacterially expressed MBD1 protein bound preferentially to methylated DNA fragments containing CpG islands from the tumor suppressor genes p16, VHL, and E cadherin and from an imprinted SNRPN gene. All MBD1 isoforms inhibited promoter activities of these genes via methylation. Interestingly, MBD1 isoforms v1 and v2 containing three CXXC domains also suppressed unmethylated promoter activities in mammalian cells. These effects were further manifested in Drosophila melanogaster cells, which lack genome methylation. Sp1-activated transcription of methylated p16 and SNRPN promoters was inhibited by all of the MBD1 isoforms, whereas the isoforms v1 and v2 reduced Sp1-activated transcription from unmethylated promoters as well. These findings suggested that the MBD1 isoforms have different roles in methylation-mediated transcriptional silencing in euchromatin. PMID- 10454588 TI - Functional analysis of H-Ryk, an atypical member of the receptor tyrosine kinase family. AB - H-Ryk is an atypical receptor tyrosine kinase which differs from other members of this family at a number of conserved residues in the activation and nucleotide binding domains. Using a chimeric receptor approach, we demonstrate that H-Ryk has impaired catalytic activity. Despite the receptor's inability to undergo autophosphorylation or phosphorylate substrates, we demonstrate that ligand stimulation of the chimeric receptor results in activation of the mitogen activated protein kinase pathway. The ability to transduce signals is abolished by mutation of the invariant lysine (K334A) in subdomain II of H-Ryk. Further, by in vitro mutagenesis, we show that the amino acid substitutions in the activation domain of H-Ryk account for the loss of catalytic activity. In addition to the essential aspartate residue, either phenylalanine or glycine is required in the activation domain to maintain proper conformation of the catalytic domain and thus ensure receptor autophosphorylation. Homology modelling of the catalytic domain of H-Ryk provides a rationale for these findings. Thus, the signalling properties of H-Ryk are divergent from those of other classical receptor tyrosine kinases. PMID- 10454589 TI - Trithorax and ASH1 interact directly and associate with the trithorax group responsive bxd region of the Ultrabithorax promoter. AB - Trithorax (TRX) and ASH1 belong to the trithorax group (trxG) of transcriptional activator proteins, which maintains homeotic gene expression during Drosophila development. TRX and ASH1 are localized on chromosomes and share several homologous domains with other chromatin-associated proteins, including a highly conserved SET domain and PHD fingers. Based on genetic interactions between trx and ash1 and our previous observation that association of the TRX protein with polytene chromosomes is ash1 dependent, we investigated the possibility of a physical linkage between the two proteins. We found that the endogenous TRX and ASH1 proteins coimmunoprecipitate from embryonic extracts and colocalize on salivary gland polytene chromosomes. Furthermore, we demonstrated that TRX and ASH1 bind in vivo to a relatively small (4 kb) bxd subregion of the homeotic gene Ultrabithorax (Ubx), which contains several trx response elements. Analysis of the effects of ash1 mutations on the activity of this regulatory region indicates that it also contains ash1 response element(s). This suggests that ASH1 and TRX act on Ubx in relatively close proximity to each other. Finally, TRX and ASH1 appear to interact directly through their conserved SET domains, based on binding assays in vitro and in yeast and on coimmunoprecipitation assays with embryo extracts. Collectively, these results suggest that TRX and ASH1 are components that interact either within trxG protein complexes or between complexes that act in close proximity on regulatory DNA to maintain Ubx transcription. PMID- 10454590 TI - A novel role for helix 12 of retinoid X receptor in regulating repression. AB - Nutrients, drugs, and hormones influence transcription during differentiation and metabolism by binding to high-affinity nuclear receptors. In the absence of ligand, some but not all nuclear receptors repress transcription as a heterodimer with retinoid X receptor (RXR). Here we define a novel role for helix 12 (H12) in sterically masking the corepressor (CoR) binding site in apo-RXR. Removing H12 converts RXR to a potent transcriptional repressor. The length but not the specific sequence of H12 is critical for masking RXR's intrinsic repression function. This contrasts with the amphipathic character required for mediating ligand-dependent activation and coactivator recruitment. Physiologically, we show that heterodimerization of RXR with apo-thyroid hormone receptor (TR) unmasks the CoR binding site in RXR and allows the TR-RXR heterodimer to repress. A molecular mechanism that involves sequence-specific interaction between RXR H12 and the coactivator-binding surface of the nuclear receptor is proposed for this heterodimerization-mediated unmasking. Peroxisome proliferator-activated receptor gamma does not interact as well with RXR H12, thus explaining its inability to repress transcription as an RXR heterodimer. The requirement to unmask RXR's latent repression function explains why only certain RXR partners repress transcription. PMID- 10454591 TI - Disruption of retinoblastoma protein family function by human papillomavirus type 16 E7 oncoprotein inhibits lens development in part through E2F-1. AB - Complexes between the retinoblastoma protein (pRb) and the transcription factor E2F-1 are thought to be important for regulating cell proliferation. We have shown previously that the E7 oncoprotein from human papillomavirus type 16, dependent upon its binding to pRb proteins, induces proliferation, disrupts differentiation, and induces apoptosis when expressed in the differentiating, or fiber, cells of the ocular lenses in transgenic mice. Mice that carry a null mutation in E2F-1 do not exhibit any defects in proliferation and differentiation in the lens. By examining the lens phenotype in mice that express E7 on an E2F-1 null background, we now show genetic evidence that E7's ability to alter the fate of fiber cells is partially dependent on E2F-1. On the other hand, E2F-1 status does not affect E7-induced proliferation in the undifferentiated lens epithelium. These data provide genetic evidence that E2F-1, while dispensible for normal fiber cell differentiation, is one mediator of E7's activity in vivo and that the requirement for E2F-1 is context dependent. These data suggest that an important role for pRb-E2F-1 complex during fiber cell differentiation is to negatively regulate cell cycle progression, thereby allowing completion of the differentiation program to occur. PMID- 10454593 TI - New yeast genes important for chromosome integrity and segregation identified by dosage effects on genome stability. AB - Phenotypes produced by gene overexpression may provide important clues to gene function. Here, we have performed a search for genes that affect chromo-some stability when overexpressed in the budding yeast Saccharomyces cerevisiae. We have obtained clones encompassing 30 different genes. Twenty-four of these genes have been previously characterized. Most of them are involved in chromatin dynamics, cell cycle control, DNA replication or mitotic chromosome segregation. Six novel genes obtained in this screen were named CST (chromosome stability). Based on the pattern of genomic instability, inter-action with checkpoint mutations and sensitivity to chromosome replication or segregation inhibitors, we conclude that overexpression of CST4 specifically interferes with mitotic chromosome segregation, and CST6 affects some aspect of DNA metabolism. The other CST genes had complex pleiotropic phenotypes. We have created deletions of five genes obtained in this screen, CST9, CST13, NAT1, SBA1 and FUN30. None of these genes is essential for viability, and deletions of NAT1 and SBA1 cause chromosome instability, a phenotype not previously associated with these genes. This work shows that analysis of dosage effects is complementary to mutational analysis of chromosome transmission fidelity, as it allows the identification of chromosome stability genes that have not been detected in mutational screens. PMID- 10454592 TI - A tale of three fingers: the family of mammalian Sp/XKLF transcription factors. AB - One of the most common regulatory elements is the GC box and the related GT/CACC box, which are widely distributed in promoters, enhancers and locus control regions of housekeeping as well as tissue-specific genes. For long it was generally thought that Sp1 is the major factor acting through these motifs. Recent discoveries have shown that Sp1 is only one of many transcription factors binding and acting through these elements. Sp1 simply represents the first identified and cloned protein of a family of transcription factors characterised by a highly conserved DNA-binding domain consisting of three zinc fingers. Currently this new family of transcription factors has at least 16 different mammalian members. Here, we will summarise and discuss recent advances that have been directed towards understanding the biological role of these proteins. PMID- 10454594 TI - An origin of bidirectional DNA replication is located within a CpG island at the 3" end of the chicken lysozyme gene. AB - We previously identified a broad initiation zone of DNA replication at the chicken lysozyme gene locus. However, the existence of a highly preferred origin of bidirectional replication (OBR), often found in initiation zones, remained elusive. In order to re-examine this issue we used a competitive PCR assay to determine the abundance of closely spaced genomic segments in a 1 kb size fraction of nascent DNA. A sharp peak of nascent strand abundance occurred at the 3" end of the gene, where initiation events were 17 times more frequent than upstream of the gene. This primary initiation site, active in lysozyme expressing myelomonocytic HD11 cells and non-expressing hepatic DU249 cells, was found to reside within an unusually located CpG island. While most CpG islands are found at the 5" end of genes, the lysozyme gene island extends from the 3" end of the second intron and includes approximately 1.2 kb of 3" flanking DNA. As diagnosed by methylation-sensitive restriction enzymes, the island is largely non methylated in HD11 cells, DU249 cells and inactive chicken erythrocytes. Furthermore, a DNase I hypersensitive site (HS) that is composed of two subsites separated by approximately 100 bp, was localised very close to the segment with the highest initiation activity. Our results suggest that the non-methylated CpG island and the HS provide an accessible chromatin structure for the lysozyme gene origin of replication. PMID- 10454595 TI - The effect of sodium, potassium and ammonium ions on the conformation of the dimeric quadruplex formed by the Oxytricha nova telomere repeat oligonucleotide d(G(4)T(4)G(4)). AB - The DNA sequence d(G(4)T(4)G(4)) [Oxy-1.5] consists of 1.5 units of the repeat in telomeres of Oxytricha nova and has been shown by NMR and X-ray crystallographic analysis to form a dimeric quadruplex structure with four guanine-quartets. However, the structure reported in the X-ray study has a fundamentally different conformation and folding topology compared to the solution structure. In order to elucidate the possible role of different counterions in this discrepancy and to investigate the conformational effects and dynamics of ion binding to G quadruplex DNA, we compare results from further experiments using a variety of counterions, namely K(+), Na(+)and NH(4)(+). A detailed structure determination of Oxy-1.5 in solution in the presence of K(+)shows the same folding topology as previously reported with the same molecule in the presence of Na(+). Both conformations are symmetric dimeric quadruplexes with T(4)loops which span the diagonal of the end quartets. The stack of quartets shows only small differences in the presence of K(+)versus Na(+)counterions, but the T(4)loops adopt notably distinguishable conformations. Dynamic NMR analysis of the spectra of Oxy-1.5 in mixed Na(+)/K(+)solution reveals that there are at least three K(+)binding sites. Additional experiments in the presence of NH(4)(+)reveal the same topology and loop conformation as in the K(+)form and allow the direct localization of three central ions in the stack of quartets and further show that there are no specific NH(4)(+)binding sites in the T(4)loop. The location of bound NH(4)(+)with respect to the expected coordination sites for Na(+)binding provides a rationale for the difference observed for the structure of the T(4)loop in the Na(+)form, with respect to that observed for the K(+)and NH(4)(+)forms. PMID- 10454597 TI - Oligonucleotide dendrimers: stable nano-structures. AB - DNA dendrimers with two, three, six, nine or 27 arms were reassociated as complementary pairs in solution or with an array of complementary oligonucleotides on a solid support. In all cases, duplex stabilities were greater than those of unbranched molecules of equal length. A theoretical treatment for the process of dissociation of dendrimers explains the major properties of the complexes. The favourable features of DNA dendrimers-their enhanced stability and the simple predictability of their association behaviour makes them promising as building blocks for the 'bottom up' approach to nano assembly. These features also suggest applications in oligonucleotide array/DNA chip technology when higher hybridisation temperatures are required, for example, to melt secon-dary structure in the target. PMID- 10454596 TI - Optimization of alternate-strand triple helix formation at the 5"-TpA-3" and 5" ApT-3" junctions. AB - Alternate-strand triple helix formation was optimized at the two junction steps, the 5"-TpA-3" and 5"-ApT-3" junctions. Footprint experiments, gel retardation assays and thermal denaturation measures on a sequence appropriately designed with two adjacent alternate-strand polypurine tracts points out that the addition of an adenine residue and the removal of one nucleotide should facilitate the crossing strands at the 5"-TpA-3" junction and at the 5"-ApT-3" junction, respectively. These results provide a 'switch code' for the construction of alternate-strand triple helix forming oligonucleotides which open new possibilities for extending the range of applications of antigene strategy. PMID- 10454598 TI - Regulation of the ribonucleotide reductase small subunit gene by DNA-damaging agents in Dictyostelium discoideum. AB - In Escherichia coli, yeast and mammalian cells, the genes encoding ribonucleotide reductase, an essential enzyme for de novo DNA synthesis, are up-regulated in response to DNA damaging agents. We have examined the response of the rnrB gene, encoding the small subunit of ribonucleotide reductase in Dictyostelium discoideum, to DNA damaging agents. We show here that the accumulation of rnrB transcript is increased in response to methyl methane sulfonate, 4-nitroquinoline 1-oxide and irradiation with UV-light, but not to the ribonucleotide reductase inhibitor hydroxyurea. This response is rapid, transient and independent of protein synthesis. Moreover, cells from different developmental stages are able to respond to the drug in a similar fashion, regardless of the basal level of expression of the rnrB gene. We have defined the cis -acting elements of the rnrB promoter required for the response to methyl methane sulfonate and 4 nitroquinoline-1-oxide by deletion analysis. Our results indicate that there is one element, named box C, that can confer response to both drugs. Two other boxes, box A and box D, specifically conferred response to methyl methane sulfonate and 4-nitroquinoline-1-oxide, respectively. PMID- 10454599 TI - RecG helicase activity at three- and four-strand DNA structures. AB - The RecG helicase of Escherichia coli is necessary for efficient recombination and repair of DNA in vivo and has been shown to catalyse the unwinding of DNA junctions in vitro. Despite these findings, the precise role of RecG remains elusive. However, models have been proposed in which RecG promotes the resolution of linked duplexes by targeting three-strand junctions present at D-loops. One such model postulates that RecG catalyses the formation of four-strand (Holliday) junctions from three-strand junctions. To test this model, the DNA binding and unwinding activities of RecG were analysed using synthetic three- and four-strand junctions. The substrate specificity of RecG was found to depend critically on the concentrations of ATP and MgCl(2)and under certain conditions RecG preferentially unwound three-strand junction DNA. This was at least partly due to the larger inhibitory effect of MgCl(2)on the binding of four-strand as opposed to three-strand junctions by RecG. Thus RecG may be targeted to three-strand junctions in vivo whilst still being able to branch migrate the four-strand junctions formed as a result of the initial helicase reaction. The increase in the dissociation constant of RecG on conversion of a three-strand into a four strand junction may also facilitate resolution of the four-strand junction by the RuvABC complex. PMID- 10454600 TI - Characterization of the interaction of lambda exonuclease with the ends of DNA. AB - Lambda exonuclease processively degrades one strand of double-stranded DNA (dsDNA) in the 5"-3" direction. To understand the mechanism through which this enzyme generates high processivity we are analyzing the first step in the reaction, namely the interaction of lambda exonuclease with the ends of substrate DNA. Endonuclease mapping of lambda exonuclease bound to DNA has shown that the enzyme protects approximately 13-14 bp on dsDNA, and no nucleo-tides on the single-stranded tail of the DNA product. We have developed a rapid fluorescence based assay using 2-aminopurine and measured the steady-state rate constants for different end-structures of DNA. The relative k(cat)for 5" ends decreases in the order 5" recessed > blunt >> 5" overhang. However, k(cat)/K(m)remains relatively constant for these different structures suggesting they are all used equally efficiently as substrates. From these data we propose that a single-stranded 5" overhang end can bind non-productively to the enzyme and the non-hydrolyzed strand is required to aid in the proper alignment of the 5" end. We have also measured the length-dependence of the steady-state rate para-meters and find that they are consistent with a high degree of processivity. PMID- 10454601 TI - Evolutionary conserved mechanism of transcriptional repression by even-skipped. AB - Even-skipped (Eve) is a transcriptional repressor involved in segment formation in Drosophila melano-gaster. In order to gain further insights into the mechanism of action of Eve we tested whether it would function as a transcriptional repressor in mammalian cells. We found that Eve was indeed a potent repressor in two different mammalian cell types and at several promoters. In vitro transcription assays confirmed that Eve directly represses transcription initiation when specifically targeted to a promoter. We also found that, unlike the case with transcriptional activators, Eve does not repress transcription synergistically. Analysis of the effect of Eve on preinitiation complex assembly in a crude HeLa cell nuclear extract demonstrated that the Eve repression domain functions by preventing the assembly of TFIID with the promoter. Our data support the hypothesis that Eve contains an active repression domain that functions specifically to prevent preinitiation complex formation. PMID- 10454602 TI - Structural equivalence in the transcribed spacers of pre-rRNA transcripts in Schizosaccharomyces pombe. AB - The structure of the internal transcribed spacer 2 (ITS2) in Schizosaccharomyces pombe was re-evaluated with respect to phylogenetically conserved features in yeasts, features in other transcribed spacer regions as well as the binding of transacting factors which potentially play a role in ribosomal maturation. Computer analyses and probes for nuclease protection indicate a very simple core structure consisting of a single extended hairpin which includes the interacting termini of the mature 5.8S and 25S rRNAs. Comparisons with ITS2 sequences in greatly diverging organisms indicate that the same feature also can be recognized. This is especially clear in organisms that contain very short sequences in which the putative structures are much less ambiguous. Diversity between organisms is the result of changes in hairpin length as well as the addition of branched helices. Protein binding and gel retardation studies with the S.pombe ITS2 further indicate that, as observed in the 3" external transcribed spacer (ETS) and ITS1 regions, the extended hairpin is not only the site of intermediate RNA cleavage during rRNA processing but also a site for specific interactions with one or more soluble factors. Taken together with other analyses on transcribed spacer regions, the present data suggest that the spacer regions all may act in a similar fashion, not only to organize the maturing terminal sequences, but also serve to organize specific soluble factors possibly acting with snoRNAs or in a manner which is analogous with that of the free snoRNPs. PMID- 10454603 TI - Long range cooperative interactions regulate the initiation of replication in the Tetrahymena thermophila rDNA minichromosome. AB - The Tetrahymena thermophila rDNA exists as a 21 kb palindromic minichromosome with two initiation sites for replication in each half palindrome. These sites localize to the imperfect, repeated 430 bp segments that include the nucleosome free domains 1 and 2 (D1 and D2). To determine if the D1 and D2 segments act independently or in concert to control initiation, stable DNA transformation assays were performed. Single domain derivatives of the plasmid prD1 failed to support autonomous replication in Tetrahymena. Instead, such constructs propagated exclusively by integration into endogenous rDNA minichromosomes and displayed weak origin activity as detected by 2D gel electrophoresis. D1/D1 and D2/D2 derivatives also transformed Tetrahymena poorly, showing similar replication defects. Hence, the D1 and D2 segments are functionally non-redundant and cooperate rather than compete to control initiation. The observed replication defect was greatly reduced in a plasmid derivative that undergoes palindrome formation in Tetrahymena, suggesting that a compensatory mechanism overcomes this replication block. Finally, using a transient replication assay, we present evidence that phylogenetically-conserved type I elements directly regulate DNA replication. Taken together, our data support a model in which cooperative interactions between dispersed elements coordinately control the initiation of DNA replication. PMID- 10454604 TI - Layer-by-layer deposition of oppositely charged polyelectrolytes on the surface of condensed DNA particles. AB - DNA can be condensed with an excess of poly-cations in aqueous solutions forming stable particles of submicron size with positive surface charge. This charge surplus can be used to deposit alternating layers of polyanions and polycations on the surface surrounding the core of condensed DNA. Using poly-L-lysine (PLL) and succinylated PLL (SPLL) as polycation and polyanion, respectively, we demonstrated layer-by-layer architecture of the particles. Polyanions with a shorter carboxyl/backbone distance tend to disassemble binary DNA/PLL complexes by displacing DNA while polyanions with a longer carboxyl/backbone distance effectively formed a tertiary complex. The zeta potential of such complexes became negative, indicating effective surface recharging. The charge stoichiometry of the DNA/PLL/SPLL complex was found to be close to 1:1:1, resembling poly-electrolyte complexes layered on macrosurfaces. Recharged particles containing condensed plasmid DNA may find applications as non-viral gene delivery vectors. PMID- 10454606 TI - Direct identification of NH...N hydrogen bonds in non-canonical base pairs of RNA by NMR spectroscopy. AB - It is shown that the recently developed quantitative J(NN)HNN-COSY experiment can be used for the direct identification of hydrogen bonds in non-canonical base pairs in RNA. Scalar(2h)J(NN)couplings across NH.N hydrogen bonds are observed in imino hydrogen bonded GA base pairs of the hpGA RNA molecule, which contains a tandem GA mismatch, and in the reverse Hoogsteen AU base pairs of the E-loop of Escherichia coli 5S rRNA. These scalar couplings correlate the imino donor(15)N nucleus of guanine or uridine with the acceptor N1 or N7 nucleus of adenine. The values of the corresponding(2h)J(NN)coupling constants are similar in size to those observed in Watson-Crick base pairs. The reverse Hoogsteen base pairs could be directly detected for the E-loop of E.coli 5S rRNA both in the free form and in a complex with the ribosomal protein L25. This supports the notion that the E loop is a pre-folded RNA recognition site that is not subject to significant induced conformational changes. Since Watson-Crick GC and AU base pairs are also readily detected the HNN-COSY experiment provides a useful and sensitive tool for the rapid identification of RNA secondary structure elements. PMID- 10454605 TI - Repair of apurinic/apyrimidinic sites by UV damage endonuclease; a repair protein for UV and oxidative damage. AB - UV damage endonuclease (UVDE) initiates a novel form of excision repair by introducing a nick imme-diately 5" to UV-induced cyclobutane pyrimidine dimers or 6-4 photoproducts. Here, we report that apurinic/apyrimidinic (AP) sites are also nicked by Neurospora crassa and Schizosaccharomyces pombe UVDE. UVDE introduces a nick immediately 5" to the AP site leaving a 3"-OH and a 5"-phosphate AP. Apyrimidinic sites are more effectively nicked by UVDE than apurinic sites. UVDE also possesses 3"-repair activities for AP sites nicked by AP lyase and for 3" phosphoglycolate produced by bleomycin. The Uvde gene introduced into Escherichia coli cells lacking two types of AP endonuclease, Exo III and Endo IV, gave the host cells resistance to methylmethane sulfonate and t-butyl hydroperoxide. We identified two AP endonuclease activities in S.pombe cell extracts. Besides cyclobutane pyrimidine dimers and 6-4 photoproducts, N. crassa UVDE also nicks Dewar photoproducts. Thus, UVDE is able to repair both of the major forms of DNA damage in living organisms: UV-induced DNA lesions and AP sites. PMID- 10454607 TI - Post-transcriptional regulation of the DNA damage-inducible gadd45 gene in human breast carcinoma cells exposed to a novel retinoid CD437. AB - The biologically active synthetic retinoid CD437 (6-[3-adamantyl-4-hydroxyphenyl] 2-naphthalene, AHPN) and different human breast carcinoma (HBC) cell lines were used to examine the possible mechanism(s) of gadd45 induction. Northern blot analysis of mRNA isolated from MCF-7, MDA-MB-468 and MDA-MB-231 HBC cell lines demonstrated a progressive increase in the 1.4 kb gadd45 transcript after exposure to 1 microM CD437. Western blot analysis showed increased gadd45 protein levels in MDA-MB-468 HBC cells following exposure to CD437. CD437 increased gadd45 mRNA levels by approximately 20-fold in MDA-MB-468 cells, however, the transcriptional activity was increased approximately 2-3-fold as demonstrated by the human gadd45 promoter-luciferase reporter construct and nuclear run-off assays. Sublines of MDA-MB-468 HBC cells expressing stably integrated GADD45 cDNA fragments were obtained and CD437-dependent induction of GADD45 analyzed. We report that approximately 300 nt located in the 5"-untranslated region (5"-UTR) of gadd45 mRNA are involved in the CD437-dependent 4-fold enhanced stability of gadd45 transcripts. MDA-MB-468 cells were stably transfected with either a plasmid having a CMV promoter-driven rabbit beta-globin gene or plasmids having a CMV promoter-driven chimeric gadd45 5"-UTR-rabbit beta-globin gene, where the entire gadd45 5"-UTR (from +1 to +298) or a 45 bp subfragment of the gadd45 5" UTR (from +10 to +55) was positioned at the 5"-end of the rabbit beta-globin gene. CD437 was found to up-regulate expression of both the chimeric gadd45 rabbit beta-globin transcripts, suggesting that cis element(s) involved in the CD437-dependent enhanced stability of gadd45 mRNA are contained in the 45 nt of the 5"-UTR of the gadd45 mRNA. PMID- 10454608 TI - Isolation and characterisation of the cDNA encoding a glycosylated accessory protein of pea chloroplast DNA polymerase. AB - The cDNA encoding p43, a DNA binding protein from pea chloroplasts (ct) that binds to cognate DNA polymerase and stimulates the polymerase activity, has been cloned and characterised. The characteristic sequence motifs of hydroxyproline rich glyco-proteins (HRGP) are present in the cDNA corres-ponding to the N terminal domain of the mature p43. The protein was found to be highly O arabinosylated. Chemically deglycosylated p43 (i.e. p29) retains its binding to both DNA and pea ct-DNA polymerase but fails to stimulate the DNA polymerase activity. The mature p43 is synthesised as a pre-p43 protein containing a 59 amino acid long transit peptide which undergoes stromal cleavage as evidenced from the post-translational in vitro import of the precursor protein into the isolated intact pea chloroplasts. Surprisingly, p43 is found only in pea chloroplasts. The unique features present in the cloned cDNA indicate that p43 is a novel member of the HRGP family of proteins. Besides p43, no other DNA polymerase accessory protein with O-glycosylation has been reported yet. PMID- 10454609 TI - The polyoma virus enhancer cannot substitute for DNase I core hypersensitive sites 2-4 in the human beta-globin LCR. AB - The polyoma virus enhancer (PyE) is capable of conferring integration position independent expression to linked genes in stably transfected erythroid cells after joining to DNase I hypersensitive site (HS) 5 of the human beta-globin locus control region (LCR). In attempting to separate the chromatin opening activity of the LCR from its enhancer activity and to investigate contributions of the individual HS core elements to LCR function, the human beta-globin LCR HS2, HS3 and HS4 core elements were replaced with the PyE within the context of a yeast artificial chromosome (YAC) bearing the whole locus. We show here that, in contrast to its function in cultured cells, the PyE is unable to replace HS core element function in vivo. We found that the PyE substitution mutant LCR is unable to provide either chromatin opening or transcriptional potentiating activity at any erythroid developmental stage in transgenic mice. These data provide direct evidence that the human beta-globin LCR core elements specify unique functions that cannot be replaced by a ubiquitous enhancer activity. PMID- 10454610 TI - Exposition of a family of RNA m(5)C methyltransferases from searching genomic and proteomic sequences. AB - The Escherichia coli fmu gene product has recently been determined to be the 16S rRNA m(5)C 967 methyltransferase. As such, Fmu represents the first protein identified as an S -adenosyl-L-methionine (AdoMet)- dependent RNA m(5)C methyltransferase whose amino acid sequence is known. Using the amino acid sequence of Fmu as an initial probe in an iterative search of completed DNA sequence databases, 27 homologous ORF products were identified as probable RNA m(5)C methyltransferases. Further analysis of sequences in undeposited genomic sequencing data and EST databases yielded more than 30 additional homologs. These putative RNA m(5)C methyltransferases are grouped into eight subfamilies, some of which are predicted to consist of direct genetic counterparts, or orthologs. The enzymes proposed to be RNA m(5)C methyltransferases have sequence motifs closely related to signature sequences found in the well-studied DNA m(5)C methyltransferases and other AdoMet-dependent methyltransferases. Structure function correlates in the known AdoMet methyltransferases support the assignment of this family as RNA m(5)C methyltransferases. PMID- 10454611 TI - Analysis of the chicken TBP-like protein(tlp) gene: evidence for a striking conservation of vertebrate TLPs and for a close relationship between vertebrate tbp and tlp genes. AB - TLP (TBP-like protein), which is a new protein dis-covered by us, has a structure similar to that of the C-terminal conserved domain (CCD) of TBP, although its function has not yet been elucidated. We isolated cDNA and genomic DNA that encode chicken TLP (cTLP) and determined their structures. The predicted amino acid sequence of cTLP was 98 and 91% identical to that of its mammalian and Xenopus counterparts, respectively, and its translation product was ubiquitously observed in chicken tissues. FISH detection showed that chicken tlp and tbp genes were mapped at 3q2.6-2.8 and 3q2.4-2.6 of the same chromosome, respectively. Genome analysis revealed that the chicken tlp gene was spliced with five introns. Interestingly, the vertebrate tbp genes were also found to be split by five introns when we focused on the CCDs, and their splicing points were similar to those of tlp. On the contrary, another TBP-resembling gene of Drosophila, trf1, is split by only one intron, as is the Drosophila 's tbp gene. These results support our earlier assumption that vertebrate TLPs did not directly descend from Drosophila TRF1. On the basis of these results together with phylogenetical exam ination, we speculate that tlp diverged from an ancestral tbp gene through a process of gene duplication and point mutations. PMID- 10454612 TI - Repair of oxidative DNA base lesions induced by fluorescent light is defective in xeroderma pigmentosum group A cells. AB - Fluorescent light (FL) has been shown to generate free radicals within cells, however, the specific chemical nature of DNA damage induced by FL has not previously been determined. Using gas chromatography/isotope dilution mass spectrometry, we have detected induction of the oxidative DNA lesions 5 hydroxycytosine (5-OH-Cyt), 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua) and 4, 6-diamino-5-formamidopyrimidine (FapyAde) in cultured cells irradiated with FL. We followed the repair of these lesions in normal and xeroderma pigmentosum group A (XP-A) cells. 5-OH-Cyt and FapyGua were repaired efficiently in normal cells within 6 h following FL exposure. XP-A cells were unable to repair these oxidative DNA base lesions. Additionally, to compare the repair of oxidative lesions induced by various sources, in vitro repair studies were performed using plasmid DNA damaged by FL, gamma-irradiation or OsO(4)treatment. Whole cell extracts from normal cells repaired damaged substrates efficiently, whereas there was little repair in XP-A extracts. Our data demon-strate defective repair of oxidative DNA base lesions in XP-A cells in vivo and in vitro. PMID- 10454613 TI - Inhibition of luciferase expression by synthetic hammerhead ribozymes and their cellular uptake. AB - Two synthetic hammerhead ribozymes, one unmodified and the other with 2" modifications and four phosphorothioate groups, targeting a single GUA site in the luciferase mRNA, were compared for their inhibition of gene expression in cell cultureand their cellular uptake was also analysed. A HeLa X1/5 cell line stably expressing luciferase, under an inducible promoter, was treated with these ribozymes by liposome-mediated transfection to determine their activity. Luciferase expression in cells was inhibited to approximately 50% with little difference between the unmodified and the 2"-modified ribozyme. A similar degree of inhibition was observed with two catalytically inactive ribozymes, indicating that inhibition was mainly due to an antisense effect. A ribozyme carrying a cholesterol moiety, applied to the cells without carrier, showed no inhibition. Northern blotting indicated a similar amount of cellular uptake of all ribozymes. The unmodified ribozyme was essentially evenly distributed between cytoplasm and nucleus, whereas a higher proportion of the phosphorothioate-containing ribozyme was observed in the nucleus. Fluorescence microscopy, including confocal microscopy using 5"-fluorescein-labelled ribozymes, showed that the unmodified and 2"-modified ribozymes were present in the cytoplasm and in the nucleus to a similar extent, whereas the fluorescence of the phosphorothioate-containing ribozyme was much stronger in the nucleus. Both ribozymes inhibited luciferase expression to a comparable degree, suggesting that the ribozyme in the nucleus did not contribute significantly to the inhibition. Ribozymes with a cholesterol moiety were predominantly trapped in the cell membrane, explaining their inability to interfere with gene expression. PMID- 10454614 TI - Faithful expression of a heterologous gene carried on an artificial macronuclear chromosome in Euplotes crassus. AB - Macronuclear chromosomes of hypotrichous ciliates are gene-sized molecules carrying the coding sequence flanked by short non-translated regions and bounded by telomeres. We have constructed artificial chromosomes for investigation of transcription in the macronucleus of Euplotes crassus. The neo gene was put under the control of the 5"-non-translated region of the TBP gene of E.crassus. These molecules were introduced into the cell with the help of liposomes. The cells were transformed and survived high concentrations of geneticin. The artificial chromosomes were kept in the macro-nucleus for at least 50 days at a copy number of about 200 per macronucleus. Expression of the gene was shown by reverse transcription of the neo messenger. The transcription start was mapped and found to coincide with that found on the natural macronuclear chromosome encoding TBP in E.crassus. PMID- 10454615 TI - Transcriptional pause, arrest and termination sites for RNA polymerase II in mammalian N- and c-myc genes. AB - Using either highly purified RNA polymerase II (pol II) elongation complexes assembled on oligo(dC)-tailed templates or promoter-initiated (extract-generated) pol II elongation complexes, the precise 3" ends of transcripts produced during transcription in vitro at several human c- and N- myc pause, arrest and termination sites were determined. Despite a low overall similarity between the entire c- and N- myc first exon sequences, many positions of pol II pausing, arrest or termination occurred within short regions of related sequence shared between the c- and N- myc templates. The c- and N- myc genes showed three general classes of sequence conservation near intrinsic pause, arrest or termination sites: (i) sites where arrest or termination occurred after the synthesis of runs of uridines (Us) preceding the transcript 3" end, (ii) sites downstream of potential RNA hairpins and (iii) sites after nucleotide addition following either a U or a C or following a combination of several pyrimidines near the transcript 3" end. The finding that regions of similarity occur near the sites of pol II pausing, arrest or termination suggests that the mechanism of c- and N- myc regulation at the level of transcript elongation may be similar and not divergent as previously proposed. PMID- 10454616 TI - Photocrosslinking locates a binding site for the large subunit of human replication protein A to the damaged strand of cisplatin-modified DNA. AB - The repair proteins XPA, XPC and replication protein A (RPA) have been implicated in the primary recognition of damaged DNA sites during nucleotide excision repair. Detailed structural information on the binding of these proteins to DNA lesions is however lacking. We have studied the binding of human RPA (hRPA) and hRPA-XPA-complexes to model oligonucleo-tides containing a single 1, 3-d(GTG) cisplatin-modification by photocrosslinking and electrophoretic mobility shift experiments. The 70 kDa subunit of hRPA can be crosslinked with high efficiency to cisplatin-modified DNA probes carrying 5-iodo-2"-deoxyuridin (5-IdU) as crosslinking chromophore. High efficiency crosslinking is dependent on the presence of the DNA lesion and occurs preferentially at its 5"-side. Examination of the crosslinking efficiency in dependence on the position of the 5-IdU chromophore indicates a specific positioning of hRPA with respect to the platination site. When hRPA and XPA are both present mainly hRPA is crosslinked to the DNA. Our mobility shift experiments directly show the formation of a stable ternary complex of hRPA, XPA and the damaged DNA. The affinity of the XPA hRPA complex to the damaged DNA is increased by more than one order of magnitude as compared to hRPA alone. PMID- 10454617 TI - Pyrophosphorolytic dismutation of oligodeoxy-nucleotides by terminal deoxynucleotidyltransferase. AB - Terminal transferase (TdT), when incubated with a purified(32)P-5"-end-labeled oligonucleotide of defined length in the presence of Co(2+), Mn(2+)or Mg(2+)and 2 mercaptoethanol in cacodylate or HEPES buffer, pH 7.2, exhibits the ability to remove a 3"-nucleotide from one oligonucleotide and add it to the 3"-end of another. When analyzed by urea-PAGE, this activity is observed as a disproportionation of the starting oligonucleotide into a ladder of shorter and longer oligonucleotides distributed around the starting material. Optimal metal ion concentration is 1-2 mM. All three metal ions support this activity with Co(2+)> Mn(2+) congruent with Mg(2+). Oligonucleotides p(dT) and p(dA) are more efficient substrates than p(dG) and p(dC) because the latter may form secondary structures. The dismutase activity is significant even in the presence of dNTP concentrations comparable to those that exist in the nucleus during the G(1)phase of the cell cycle. Using BetaScope image analysis the rate of pyrophosphorolytic dismutase activity was found to be only moderately slower than the poly merization activity. These results may help explain the GC-richness of immunoglobulin gene segment joins (N regions) and the loss of bases that occur during gene rearrangements in pre-B and pre-T cells. PMID- 10454618 TI - Substrate recognition by Escherichia coli MutY using substrate analogs. AB - The Escherichia coli adenine glycosylase MutY is involved in the repair of 7,8 dihydro-8-oxo-2"-deoxyguanosine (OG):A and G:A mispairs in DNA. Our approach toward understanding recognition and processing of DNA damage by MutY has been to use substrate analogs that retain the recognition properties of the substrate mispair but are resistant to the glycosylase activity of MutY. This approach provides stable MutY-DNA complexes that are amenable to structural and biochemical characterization. In this work, the interaction of MutY with the 2" deoxyadenosine analogs 2"-deoxy-2"-fluoroadenosine (FA), 2"-deoxyaristeromycin (R) and 2"-deoxyformycin A (F) was investigated. MutY binds to duplexes containing the FA, R or F analogs opposite G and OG within DNA with high affinity; however, no enzymatic processing of these duplexes is observed. The specific nature of the interaction of MutY with an OG:FA duplex was demonstrated by MPE-Fe(II) hydroxyl radical footprinting experiments which showed a nine base pair region of protection by MutY surrounding the mispair. DMS footprinting experiments with an OG:A duplex revealed that a specific G residue located on the OG-containing strand was protected from DMS in the presence of MutY. In contrast, a G residue flanking the substrate analogs R, F or FA was observed to be hypersensitive to DMS in the presence of MutY. These results suggest a major conformational change in the DNA helix upon binding of MutY that exposes the substrate analog-containing strand. This finding is consistent with a nucleotide flipping mechanism for damage recognition by MutY. This work demonstrates that duplex substrates for MutY containing FA, R or F instead of A are excellent substrate mimics that may be used to provide insight into the recognition by MutY of damaged and mismatched base pairs within DNA. PMID- 10454619 TI - Specificity of DNA binding of the c-Myc/Max and ARNT/ARNT dimers at the CACGTG recognition site. AB - Basic helix-loop-helix proteins that interact with the DNA recognition site CACGTG include the c-Myc/Max heterodimer and the ARNT (Ahreceptornucleartranslocator) homodimer. We have utilized a PCR-based protocol to identify high affinity binding sites of either the c-Myc/Max or ARNT/ARNT dimers and analyzed the ability of these dimers to interact with their derived consensus sequences and activate genes. chi(2)analysis of the selected DNA recognition sites revealed that DNA binding of the ARNT homodimer is symmetric, resulting in the consensus sequence RTCACGTGAY. Gel shift analysis demonstrated that the flanking nucleotides play an important role in dictating DNA binding affinity of the ARNT homodimer. These flanking sequences also regulate the ability of ARNT to competitively displace the c-Myc/Max heterodimer from a CACGTG containing sequence. However, transient transfection analyses in CV-1 cells revealed that ARNT and c-Myc/Max exhibited similar abilities to activate transcription through each other's consensus sequences. Taken together, these results indicate that although binding affinity of these dimers for the CACGTG core sequences may be differentially influenced by flanking nucleotides, transcriptional activity may also be determined by other factors, such as cellular concentrations of these proteins and their co-activators. PMID- 10454620 TI - Effect of oxidative DNA damage in promoter elements on transcription factor binding. AB - Reactive oxygen species produced by endogenous metabolic activity and exposure to a multitude of exogenous agents impact cells in a variety of ways. The DNA base damage 8-oxodeoxyguanosine (8-oxodG) is a prominent indicator of oxidative stress and has been well-characterized as a premutagenic lesion in mammalian cells and putative initiator of the carcinogenic process. Commensurate with the recent interest in epigenetic pathways of cancer causation we investigated how 8-oxodG alters the interaction between cis elements located on gene promoters and sequence-specific DNA binding proteins associated with these promoters. Consensus binding sequences for the transcription factors AP-1, NF-kappaB and Sp1 were modified site-specifically at guanine residues and electrophoretic mobility shift assays were performed to assess DNA-protein interactions. Our results indicate that whereas a single 8-oxodG was sufficient to inhibit transcription factor binding to AP-1 and Sp1 sequences it had no effect on binding to NF-kappaB, regardless of its position. We conclude from these data that minor alterations in base composition at a crucial position within some, but not all, promoter elements have the ability to disrupt transcription factor binding. The lack of inhibition by damaged NF-kappaB sequences suggests that DNA-protein contact sites may not be as determinative for stable p50 binding to this promoter as other, as yet undefined, structural parameters. PMID- 10454621 TI - Intron-exon structures of eukaryotic model organisms. AB - To investigate the distribution of intron-exon structures of eukaryotic genes, we have constructed a general exon database comprising all available intron containing genes and exon databases from 10 eukaryotic model organisms: Homo sapiens, Mus musculus, Gallus gallus, Rattus norvegicus, Arabidopsis thaliana, Zea mays, Schizosaccharomyces pombe, Aspergillus, Caenorhabditis elegans and Drosophila. We purged redundant genes to avoid the possible bias brought about by redundancy in the databases. After discarding those questionable introns that do not contain correct splice sites, the final database contained 17 102 introns, 21 019 exons and 2903 independent or quasi-independent genes. On average, a eukaryotic gene contains 3.7 introns per kb protein coding region. The exon distribution peaks around 30-40 residues and most introns are 40-125 nt long. The variable intron-exon structures of the 10 model organisms reveal two interesting statistical phenomena, which cast light on some previous speculations. (i) Genome size seems to be correlated with total intron length per gene. For example, invertebrate introns are smaller than those of human genes, while yeast introns are shorter than invertebrate introns. However, this correlation is weak, suggesting that other factors besides genome size may also affect intron size. (ii) Introns smaller than 50 nt are significantly less frequent than longer introns, possibly resulting from a minimum intron size requirement for intron splicing. PMID- 10454622 TI - Confluence-induced alterations in CpG island methylation in cultured normal human fibroblasts. AB - Growth constraint of bacterial and human cells has been shown to trigger genetic mutation. We questioned whether growth constraint might also trigger epigenetic mutation in the form of CpG island methylation. Logarithmically growing normal human fibro-blasts (NHF) displayed little (0-15%) CpG methylation in select regions of three CpG islands [estrogen receptor (ER), E-cadherin (ECAD) and O (6) methylguanine-DNA methyltransferase (MGMT)] examined. NHF grown to and left at confluence for 2-21 days showed little (<10%) CpG methylation in the ER and ECAD CpG islands. These confluent, growth-arrested cells, however, displayed extensive ( approximately 50%) methylation of the MGMT CpG island. CpG methylation in the MGMT CpG island was not associated with cellular senescence. The methylation was, however, heritable, but not permanent, as the level of CpG methylation in the MGMT CpG island of cells 4 population doublings following replating after confluence were no different from those in confluent cultures, but returned to levels noted in logarithmically growing cells by 10 population doublings following replating. These results suggest that growth constraint can trigger transient epigenetic change even in normal non-senescent human cells. PMID- 10454624 TI - Overcoming a barrier for DNA polymerization in triplex-forming sequences. AB - Folded structures in the DNA template, such as hairpins and multi-stranded structures, often serve as pause and arrest sites for DNA polymerases. DNA polymerization is particularly difficult on mirror-repeated homopurine.homopyrimidine templates where triple-stranded (triplex) structures may form between the nascent and folded template strands. In order to use a linear PCR amplification approach for the structural analysis of DNA in mirror repeated sequences we modified a conventional protocol. The barrier for DNA synthesis can be eliminated using an oligonucleotide that hybridizes with the template to prevent its folding and is subsequently displaced by the progressing polymerase. The described approach is potentially useful for sequencing and analysis of chemical adducts and point mutations in a variety of sequences prone to the formation of folded structures, such as long hairpins and quadruplexes. PMID- 10454623 TI - Presence of a coordinated metal ion in a trans-acting antigenomic delta ribozyme. AB - We have investigated the cleavage induced by metal ions in an antigenomic form of a trans-acting delta ribozyme. A specific Mg(2+)-induced cleavage at position G(52)at the bottom of the P2 stem was observed to occur solely within catalytically active ribozyme-substrate complexes (i.e. those that performed the essential conformational transition step). Only the divalent cations which support catalytic activity permitted the detection of specific induced cleavages in this region. Using various mutant ribozymes and substrates, we demonstrated a correlation between enzymatic activity and the Mg(2+)-induced cleavage pattern. We show that the efficiency of the coordination of the magnesium to its binding site is related to the nature of the base pair in the middle of the P1 stem (i.e. Rz(23)-S(8)). Together with additional evidence from nuclease probing experiments that indicates the occurrence of a structural rearrangement involving the bottom of the P2 stem upon formation of the P1 helix, these results show that an intimate relationship exists between the folding and the catalytic activity of the delta ribozyme. PMID- 10454625 TI - Prevention of depurination during elution facilitates the reamplification of DNA from differential display gels. AB - DNA fragments that show a pattern of differential expression on differential display gels must be eluted from the gel matrix and reamplified to enable further analysis. Elution is usually achieved by heating excised gel slices in a small volume of either water or TE. Here we show that this elution step can adversely affect the ability of the eluted DNA to act as a template for PCR reamplification, probably via the process of depurination. Simply switching to an elution solvent designed to minimise depurination (PCR buffer) facilitates the elution of intact DNA fragments. This improvement is likely to be most beneficial when eluting higher molecular weight fragments (e.g. those >500 bp), in situations where the amount of DNA in an excised gel slice is limited or when contaminating differential display products co-migrate with the differentially expressed product. PMID- 10454626 TI - Enzymatic mutation detection: enrichment of heteroduplexes from hybrid DNA mixtures by cleavage-deficient GST-tagged endonuclease VII. AB - A method for the enrichment of heteroduplex DNAs from hybrid DNA mixtures by endonuclease VII is reported. The procedure is based on the ability of a GST fused cleavage-deficient mutant endonuclease VII (EVII-N62D(GST)) to bind to mismatching nucleotides in heteroduplex DNAs identical to the wild-type enzyme. The GST tag was used for stable immobilisation of the protein to Glutathione Sepharose 4B. This enables the material to withstand the repeated rounds of binding steps required for enrichment of heteroduplex molecules from appropriate samples. PMID- 10454627 TI - Identification of differentially expressed 5'-end mRNA variants by an improved RACE technique (PEETA). AB - A rapid and efficient procedure is described for mapping and cloning the 5'-ends of mRNAs, including those generated from a unique gene by alternative splicing and promoter usage. This method involves reverse transcription of the targeted mRNAs from a long, highly labeled specific primer, resolution of the extension products on a DNA sequencing gel, elution and poly(dC) tailing of the single stranded cDNAs of interest, amplification of these cDNAs by PCR using an oligo(dG) adapter-primer and a gene-specific primer and finally DNA sequencing of the subcloned PCR fragments. The overall method is called PEETA (primer extension, electrophoresis, elution, tailing and amplification). PMID- 10454628 TI - Improved mutation detection in GC-rich DNA fragments by combined DGGE and CDGE. AB - Denaturing gradient gel electrophoresis (DGGE) has proven to be a powerful pre screening method for the detection of DNA variants. If such variants occur, however, in DNA fragments that are very rich in G and C, they may escape detection. To overcome this limitation, we tested a novel gel system which combines DGGE and constant denaturant gel electrophoresis (CDGE), as it might have the advantages of both methods. Indeed, this combination had the advantages of both methods, good separation of hetero-duplex molecules and prevention of total strand dissociation, and it proved successful in the detection of DNA variants in several GC-rich fragments. PMID- 10454629 TI - Simultaneous Cre catalyzed recombination of two alleles to restore neomycin sensitivity and facilitate homozygous mutations. AB - Cells homozygous for neo-expressing mutations can be derived by culturing heterozygotes with elevated G418. We demonstrate that this strategy is significantly less efficient if hyg is substituted for neo. Therefore, to introduce additional mutations Cre recombinase was used to remove floxed neo from both alleles of homozygotes at two different loci. The rate-determining step in Cre excision appeared independent of substrate copy number. Incorporating cytosine deaminase and Herpes simplex virus thymidine kinase allowed negative selection for both targeting and Cre excision. The resulting G418-sensitive homozygous mutants should allow mutagenesis at additional loci and avoid untoward effects of retained selection markers. PMID- 10454630 TI - A chicken embryo protein related to the mammalian DEAD box protein p68 is tightly associated with the highly purified protein-RNA complex of 5-MeC-DNA glycosylase. AB - We have shown previously that DNA demethylation by chick embryo 5-methylcytosine (5-MeC)-DNA glycosylase needs both protein and RNA. Amino acid sequences of nine peptides derived from a highly purified 5-MeC-DNA glycosylase complex were identified by Nanoelectrospray ionisation mass spectrometry to be identical to the mammalian nuclear DEAD box protein p68 RNA helicase. Antibodies directed against human p68 helicase cross-reacted with the purified 5-MeC-DNA glycosylase complex and immunoprecipitated the glycosylase activity. A 2690 bp cDNA coding for the chicken homologue of mammalian p68 was isolated and sequenced. Its derived amino acid sequence is almost identical to the human p68 DEAD box protein up to amino acid position 473 (from a total of 595). This sequence contains all the essential conserved motifs from the DEAD box proteins which are the ATPase, RNA unwinding and RNA binding motifs. The rest of the 122 amino acids in the C terminal region rather diverge from the human p68 RNA helicase sequence. The recombinant chicken DEAD box protein expressed in Escherichia coli cross-reacts with the same p68 antibodies as the purified chicken embryo 5-MeC-DNA glycosylase complex. The recombinant protein has an RNA-dependent ATPase and an ATP-dependent helicase activity. However, in the presence or absence of RNA the recombinant protein had no 5-MeC-DNA glycosylase activity. In situ hybridisation of 5 day-old chicken embryos with antisense probes of the chicken DEAD box protein shows a high abundance of its transcripts in differentiating embryonic tissues. PMID- 10454631 TI - Conversion of a DNA ligase into an RNA capping enzyme. AB - In eukaryotes, newly synthesised mRNA is 'capped' by the addition of GMP to the 5" end by RNA capping enzymes. Recent structural studies have shown that RNA capping enzymes and DNA ligases have similar protein folds, suggesting a conserved catalytic mechanism. To explore these similarities we have produced a chimeric enzyme comprising the N-terminal domain 1 of a DNA ligase fused to the C terminal domain 2 of a mRNA capping enzyme. This report shows that this hybrid enzyme retains adenylation activity, characteristic of DNA ligases but, remarkably, the chimera has ATP-dependent mRNA capping activity. This is the first observation of ATP-dependent RNA capping. These results suggest that nucleotidyltransferases may have evolved from a common ancestral gene. PMID- 10454632 TI - Excision repair of nitrogen mustard-DNA adducts in Saccharomyces cerevisiae. AB - The bifunctional alkylating anticancer drug nitrogen mustard forms a variety of DNA lesions, including monoadducts and intrastrand and interstrand crosslinks. Although it is known that nucleotide excision repair (NER) is important in processing these adducts, the role of the other principal excision repair pathway, base excision repair (BER) is less well defined. Using isogenic Saccharomyces cerevisiae strains disrupted for a variety of NER and BER genes we have examined the relative importance of the two pathways in the repair of nitrogen mustard adducts. As expected, NER defective cells (rad4 and rad14 strains) are extremely sensitive to the drug. One of the BER mutants, a 3 methyladenine glycosylase defective (mag1) strain also shows significant hypersensitivity. Using a rad4/mag1 double mutant it is shown that the two excision repair pathways are epistatic to each other for nitrogen mustard sensitivity. Furthermore, both rad14 and mag1 disruptants show elevated levels of nitrogen mustard-induced forward mutation. Measurements of repair rates of nitrogen mustard N-alkylpurine adducts in the highly transcribed RPB2 gene demonstrate defects in the processing of mono-adducts in rad4, rad14 and mag1 strains. However, there are differences in the kinetics of adduct removal in the NER mutants compared to the mag1 strain. In the mag1 strain significant repair occurs within 1 h with evidence of enhanced repair on the transcribed strand. Adducts however accumulate at later times in this strain. In contrast, in the NER mutants repair is only evident at times greater than 1 h. In a mag1/rad4 double mutant damage accumulates with no evidence of repair. Comparison of the rates of repair in this gene with those in a different genomic region indicate that the contributions of NER and BER to the repair of nitrogen mustard adducts may not be the same genome wide. PMID- 10454633 TI - The identification of nuclear proteins that bind the homopyrimidine strand of d(GA.TC)n DNA sequences, but not the homopurine strand. AB - Alternating d(GA.TC)(n)DNA sequences, which are abundant in eukaryotic genomes, can form altered DNA structures. Depending on the environmental conditions, the formation of (GA.GA) hairpins or [C+T(GA.TC)] and [GA(GA.TC)] intramolecular triplexes was observed in vitro. In vivo, the formation of these non-B-DNA structures would likely require the contribution of specific stabilizing factors. Here, we show that Friend's nuclear extracts are rich in proteins which bind the pyrimidine d(TC)(n)strand but not the purine d(GA)n strand (NOGA proteins). Upon chromatographic fractionation, four major proteins were detected (NOGA1-4) that have been purified and characterized. Purified NOGAs bind single-stranded d(TC)n with high affinity and specificity, showing no significant affinity for either d(GA)n or d(GA.TC)nDNA sequences. We also show that NOGA1, -2 and -3, which constitute the three most abundant and specific NOGA proteins, correspond to the single-stranded nucleic acid binding proteins hnRNP-L, -K and -I, respectively. These results are discussed in the context of the possible contribution of the NOGA proteins to the stabilization of the (GA.GA) and [GA(GA.TC)] conformers of the d(GA.TC)n DNA sequences. PMID- 10454634 TI - The role of nucleotide excision repair in protecting embryonic stem cells from genotoxic effects of UV-induced DNA damage. AB - In this study the role of nucleotide excision repair (NER) in protecting mouse embryonic stem (ES) cells against the genotoxic effects of UV-photolesions was analysed. Repair of cyclobutane pyrimidine dimers (CPD) in transcribed genes could not be detected whereas the removal of (6-4) photoproducts (6-4PP) was incomplete, already reaching its maximum (30%) 4 h after irradiation. Measurements of repair replication revealed a saturation of NER activity at UV doses >5 J/m2 while at a lower dose (2.5 J/m2) the repair kinetics were similar to those in murine embryonic fibroblasts (MEFs). Cytotoxic and mutagenic effects of photolesions were determined in ES cells differing in NER activity. ERCC1 deficient ES cells were hypermutable (10-fold) compared to wild-type cells, indicating that at physiologically relevant doses ES cells efficiently remove photolesions. The effect of the NER deficiency on cytoxicity was only 2-fold. Exposure to high UV doses (10 J/m2) resulted in a rapid and massive induction of apoptosis. Possibly, to avoid the accumulation of mutated cells, ES cells rely on the induction of a strong apoptotic response with a simultaneous shutting down of NER activity. PMID- 10454635 TI - Facile characterization of translation initiation via nonsense codon suppression. AB - A new strategy for studying the mechanism of translation initiation in eukaryotes has been developed. The strategy involves the use of an in vitro translation system to incorporate a non-natural fluorescent amino acid into a protein from a suppressor tRNAPheCUA misacylated with that amino acid. It is thereby possible to monitor translation initiation efficiency at an AUG codon in different contexts; this is illustrated for three constructs encoding Escherichia coli dihydrofolate reductase mRNA with different translation initiation regions. Fluorescence measurements after in vitro translation of the mRNAs in rabbit reticulocyte lysate reflected differences in the position and efficiency of translation initiation and, therefore, can be used for characterization of the translation initiation process. PMID- 10454636 TI - Dopamine stimulates expression of the human immunodeficiency virus type 1 via NF kappaB in cells of the immune system. AB - Recent studies have reported that lymphocytes produce, transport and bind dopamine present in plasma. However, the action of dopamine on HIV-1 gene expression in cells of the immune system has not yet been examined. Here, we have investigated the regulation of HIV-1 expression by dopamine in Jurkat T cells and in primary blood mononuclear cells (PBMC). HIV-1 replication was increased by dopamine, which correlated with the increased levels of HIV-1 transactivation. Our transient expression data revealed that dopamine stimulated transcription through the NF-kappaB element present in the long terminal repeat. The importance of NF-kappaB sites was confirmed by using vectors containing wild-type or mutant kappaB sites in a heterologous promoter. Consistent with the role of NF-kappaB in mediating dopamine responsiveness, the proteasome inhibitor MG132 abolished dopamine-induced transcriptional activation. We further explored the effect of dopamine in the presence of phorbol esters or tumor necrosis factor-alpha (TNF alpha) known to activate NF-kappaB. The combination of dopamine and TNF-alpha led to a stimulation of HIV-1 transcription and replication. However, in contrast with TNF-alpha, dopamine treatment did not affect NF-kappaB DNA binding activity nor the concentrations of p50, p65 and IkappaB-alpha proteins, which suggests a distinct NF-kappaB activation mechanism. These results reveal a new link between the dopamine system, cytokine signaling pathway and regulation of gene expression via the involvement of NF-kappaB in T cells and PBMC. PMID- 10454637 TI - Two 5'-ETS regions implicated in interactions with U3 snoRNA are required for small subunit rRNA maturation in Trypanosoma brucei. AB - Early pre-rRNA processing events were examined in the ancient protozoan parasite Trypanosoma brucei and found to have both distinctive and conserved features. Two 5'-ETS cleavages occur: A' and the newly discovered A0. A' and A0 appear related to vertebrate and yeast primary pre-RNA cleavage sites, respectively. However, trypanosomatid primary rRNA transcripts can first be processed at the ITS1/5.8S boundary and 5'-ETS sequences then removed by consecutive cleavages at A', A0 and A1 at the 5'-ETS/SSU rRNA junction. 5'-ETS sequences previously crosslinked to U3 snoRNA were tested for their roles in rRNA processing using our new tagged rRNA system. Two distinct A'-adjacent sequence elements, which may pair with U3 hinge bases, were specifically required for SSU rRNA production, as was a downstream element. The latter element appears conserved with the yeast 5'-ETS U3 binding sequence, required for A0, A1 and A2 cleavages, in that they both share 10 bases complementary with U3 hinge sequences and lie upstream from A0 and A1 sites located in a potential stem-loop structure. The distinctive positioning of putative trypanosomatid U3 binding sites with respect to A" and A0 cleavages suggests that different U3-dependent mechanisms may direct each processing event. PMID- 10454638 TI - Site-directed mutagenesis of motif III in PcrA helicase reveals a role in coupling ATP hydrolysis to strand separation. AB - Motif III is one of the seven protein motifs that are characteristic of superfamily I helicases. To investigate its role in the helicase mechanism we have introduced a variety of mutations at three of the most conserved amino acid residues (Q254, W259 and R260). Biochemical characterisation of the resulting proteins shows that mutation of motif III affects both ATP hydrolysis and single stranded DNA binding. We propose that amino acid residue Q254 acts as a gamma phosphate sensor at the nucleotide binding pocket transmitting conformational changes to the DNA binding site, since the nature of the charge on this residue appears to control the degree of coupling between ATPase and helicase activities. Residues W259 and R260 both participate in direct DNA binding interactions that are critical for helicase activity. PMID- 10454639 TI - Centromeres from telomeres? The centromeric region of the Y chromosome of Drosophila melanogaster contains a tandem array of telomeric HeT-A- and TART related sequences. AB - Cytological and cytogenetic studies have previously defined the region needed for centromeric function in the Y chromosome of Drosophila melanogaster. We have identified a YAC clone that originated from this region. Molecular analysis of the YAC and genomic DNAs has allowed the description of a satellite DNA made of telomeric HeT-A- and TART-derived sequences and the construction of a long-range physical map of the heterochromatic region h18. Sequences within the YAC clone are conserved in the centromeric region of the sibling species Drosophila simulans. That telomere-derived DNA now forms part of the centromeric region of the Y chromosome could indicate a telomeric origin of this centromere. The existence of common determinants for the function of both centromeres and telomeres is discussed. PMID- 10454641 TI - Novel cationic amphiphiles as delivery agents for antisense oligonucleotides. AB - There has been great interest recently in therapeutic use of nucleic acids including genes, ribozymes and antisense oligonucleotides. Despite recent improvements in delivering antisense oligonucleotides to cells in culture, nucleic acid-based therapy is still often limited by the poor penetration of the nucleic acid into the cytoplasm and nucleus of cells. In this report we describe nucleic acid delivery to cells using a series of novel cationic amphiphiles containing cholic acid moieties linked via alkylamino side chains. We term these agents 'molecular umbrellas' since the cationic alkylamino chains provide a 'handle' for binding of nucleic acids, while the cholic acid moieties are likely to interact with the lipid bilayer allowing the highly charged nucleic acid backbone to traverse across the cell membrane. Optimal gene and oligonucleotide delivery to cells was afforded by a derivative (amphiphile 5) containing four cholic acid moieties. With this amphiphile used as a constituent in cationic liposomes, a 4-5 log increase in reporter gene delivery was measured. This amphiphile used alone provided a 250-fold enhancement of oligo-nucleotide association with cells as observed by flow cytometry. A substantial fraction of cells exposed to complexes of amphiphile 5 and fluorescent oligo-nucleotide showed nuclear accumulation of the fluorophore. Enhanced pharmacological effectiveness of antisense oligonucleotides complexed with amphiphile 5 was observed using an antisense splicing correction assay that activates a Luciferase reporter. Intracellular delivery, nuclear localization and pharmacological effectiveness of oligonucleotides using amphiphile 5 were similar to those afforded by commercial cytofectins. However, in contrast to most commercial cytofectins, the umbrella amphiphile showed substantial delivery activity even in the presence of high concentrations of serum. PMID- 10454642 TI - Sequence effects on energetic and structural properties of phosphorothioate DNA: a molecular modelling study. AB - Phosphorothioate (PS) oligonucleotides constitute a new class of potent drugs, resulting from the replacement of one anionic oxygen of the phosphodiester backbone by one sulphur atom. This replacement confers chirality to the phosphorus atom (PSS or PSR) and alters the energetic, structural and biological properties of B-DNA. These properties were assessed by molecular mechanics calculations on a set of regular sequences, d(YR)8.d(YR)8 and d(RR)8.d(YY)8 (R, purine; Y, pyrimidine). Results indicated: (i) destabilisation of both the PS(R)and the PSS oligomers, the loss of total energy being mainly due to a variation in the electrostatic term; (ii) an additional chirality effect, due to van der Waals and backbone angle energies, larger for PSS oligomers than for PSR oligomers; (iii) a clear sequence effect on stability, particularly from the base immediately preceding the PS group. Even though the PS group alters the stability of oligomers, it does not significantly modify the conformation. Altogether, our molecular modelling data parallel the available experimental data. Our results reveal that sequence effects on the energetic properties of PS oligomers are local and additive. Therefore, studies of the set of the 10 unique double stranded modified dinucleotide steps included in regular oligomers could be used to predict the behaviour of any double-stranded PS-DNA. PMID- 10454640 TI - Conservation, localization and expression of HopZ, a protein involved in adhesion of Helicobacter pylori. AB - From a sarkosyl-insoluble outer membrane fraction prepared from the Helicobacter pylori strain ATCC 43504, 19 proteins could be sequenced N-terminally by Edman degradation. Oligonucleotides were deduced and used for screening of a genomic library. From the isolated genes, five code for different members of a H.pylori outer membrane protein (Hop) family. Among these, the hopZ gene was characterized in more detail. It encodes a protein which was shown to be located at the bacterial surface by immunofluorescence studies. Sequence analysis of the hopZ gene from 15 different H.pylori strains revealed the existence of two alleles and the possible regulation of hopZ expression by slipped-strand mispairing within a CT dinucleotide repeat motif located in the signal-peptide coding region. Among the different strains, the influence of this region on the expression of HopZ was analyzed on a translational level by western blot analysis of bacterial extracts and immunofluorescence studies on intact cells. The protein is expressed only in those strains in which the number of the CT dinucleotide repeats allow for an open reading frame encoding the complete protein. Addionally the function of HopZ was investigated in an adhesion assay. The wild-type strain ATCC 43504 adhered to human gastric epithel cells whereas a knockout mutant strain showed significantly reduced binding to the cells. PMID- 10454643 TI - Mammalian mitochondrial extracts possess DNA end-binding activity. AB - Mammalian mitochondrial protein extracts possess DNA end-binding (DEB) activity. Protein binding to a 394 bp double-stranded DNA molecule was measured using an electrophoretic mobility shift assay. Mitochondrial DEB activity was highly specific for linear DNA. Inclusion of a vast excess of non-radioactive circular DNA did not disrupt binding to radioactive f394. In contrast, binding was abolished by the inclusion of linear competitor DNA. In mammals, nuclear DEB activity is due to Ku, a hetero-dimer composed of the Ku70 and Ku86 proteins. To determine whether mitochondrial DEB activity was also due to Ku, protein extracts were prepared from the Chinese hamster XR-V15B cell line, which lacks this protein. As anticipated, nuclear extracts prepared from these cells lacked DEB activity. In contrast, mitochondrial extracts prepared from these cells had wild type levels of DEB activity, demonstrating that this latter activity is not a consequence of nuclear contamination. Although the nuclear and mitochondrial DEB activities are independent of each other, they are nevertheless closely related, since mitochondrial DEB activity was 'supershifted' by both anti-Ku70 and anti Ku86 antisera. The nuclear DEB protein Ku plays an essential role in nuclear DNA double-strand break repair. The DEB activity described herein may therefore play a similar role in mitochondrial DNA repair. PMID- 10454644 TI - Overproduction of histone H1 variants in vivo increases basal and induced activity of the mouse mammary tumor virus promoter. AB - BALB/c 3T3 cell lines containing integrated copies of the MMTV promoter driving a reporter gene were constructed. Expression vectors in which either of two H1 variants, H10 or H1c, were under control of an inducible promoter were introduced into these lines. Surprisingly, overproduction of either variant resulted in a dramatic increase in basal and hormone-induced expression from the MMTV promoter. H1 overproduction also slowed the loss of MMTV promoter activity associated with prolonged hormone treatment. Transiently transfected MMTV reporter genes, which do not adopt a phased nucleosomal arrangement, do not display increased activity upon H1 overproduction. Thus the effects observed for stable constructs most likely represents a direct effect of H1 on a chromatin-mediated process specific to the nucleosomal structure of the integrated constructs. Induction of increased levels of acetylated core histones by treatment with trichostatin A also potentiated MMTV activity and this effect was additive to that caused by H1 overproduction. However, the effects of TSA treatment, in control or H1 overproducing cells, were eliminated by inhibiting protein synthesis. TSA treatment does not necessarily potentiate MMTV promoter activity by increasing core histone acetylation within the MMTV promoter but perhaps by altering the synthesis of an unlinked transcriptional regulator. PMID- 10454645 TI - Heterochromatic silencing of Drosophila heat shock genes acts at the level of promoter potentiation. AB - In a variety of organisms, genes placed near heterochromatin are transcriptionally silenced. In order to understand the molecular mechanisms responsible for this block in transcription, high resolution in vivo chromatin structure analysis was performed on two heat shock genes, hsp26 and hsp70. These genes normally reside in euchromatin where GAGA factor and RNA Pol II are present on the promoter prior to heat shock induction. P-element transformation experiments led to the identification of stocks in which these two genes were inserted within heterochromatin of the chromosome 4 telomeric region. These transgenes exhibit silencing that is partially suppressed by mutations in the gene encoding HP1. Micrococcal nuclease analysis revealed that the heterochromatic transgenes were packaged in a more regular nucleosome array than when located in euchromatin. High resolution DNase I analysis demonstrated that GAGA factor and TFIID were not associated with these promoters in heterochromatin; potassium permanganate experiments showed a loss of Pol II association. Taken together, these data suggest that occlusion of trans-acting factors from their cis- acting regulatory elements leading to a block in promoter potentiation is a mechanism for heterochromatin gene silencing. PMID- 10454646 TI - Evidence from CD spectra and melting temperatures for stable Hoogsteen-paired oligomer duplexes derived from DNA and hybrid triplexes. AB - The pyr*pur.pyr type of nucleic acid triplex has a purine strand that is Hoogsteen-paired with a parallel pyrimidine strand (pyr*pur pair) and that is Watson-Crick-paired with an antiparallel pyrimidine strand (pur.pyr pair). In most cases, the Watson-Crick pair is more stable than the Hoogsteen pair, although stable formation of DNA Hoogsteen-paired duplexes has been reported. Using oligomer triplexes of repeating d(AG)12 and d(CT)12 or r(CU)12 sequences that were 24 nt long, we found that hybrid RNA*DNA as well as DNA*DNA Hoogsteen paired strands of triplexes can be more stable than the Watson-Crick-paired strands at low pH. The structures and relative stabilities of these duplexes and triplexes were evaluated by circular dichroism (CD) spectroscopy and UV absorption melting studies of triplexes as a function of pH. The CD contributions of Hoogsteen-paired RNA*DNA and DNA*DNA duplexes were found to dominate the CD spectra of the corresponding pyr*pur.pyr triplexes. PMID- 10454647 TI - Silk worm Bm1 SINE RNA increases following cellular insults. AB - The effect of cell stresses upon the expression of the Bm1 short interspersed element (SINE) family in cultured silk worm cells is examined. Primer extension analysis shows that Bm1 repeats are transcribed by RNA polymerase III (Pol III) into cytoplasmic RNAs. Five consecutive T residues, which would normally terminate Pol III transcription, occur within the Bm1 consensus and are included within cDNA sequences representing these transcripts. In analogy to mammalian SINEs, the level of the Bm1 transcripts increases in response to either heat shock, inhibiting protein synthesis by cycloheximide or viral infection. The lifetime of Bm1 RNA increases following cell insults so that post-transcriptional events partially account for stress induced increases in its abundance. In the case of heat shock, the increase in Bm1 RNA follows the transient increase in hsp70 mRNA indicating that this response is temporally regulated to occur later in heat shock recovery. These results support the proposal that SINE RNAs serve a role in the cell stress response that predates the divergence of insects and mammals implying that SINEs are essentially a class of cell stress genes. PMID- 10454648 TI - Rapid purification of intact minichromosomes over a glycerol cushion. AB - Negatively supercoiled plasmids can be assembled into dynamic minichromosomes using Drosophila embryo extract as a source of histones and chromatin assembly factors. However, analysis of such mini-chromosomes is often difficult due to the presence in the crude extract of a large excess of macromolecules and low molecular weight molecules including ATP. Several techniques have been used to partially purify the minichromosomes based on either sizing columns or centrifugation on sucrose gradients. We have developed a single-step method employing a 30 min ultracentrifugation through a glycerol cushion. In contrast to chromatin purified in sucrose-containing buffers, the minichromosomes obtained with this method are suitable for transcriptional analysis. This method is fast, quantitative, flexible, can deal with several samples simultaneously and leads to concentration of the chromatin. As centrifugation through glycerol yields chromatin free of ATP and several characterized chromatin remodeling complexes, this method should be useful for structural and functional studies in vitro. PMID- 10454649 TI - Optimisation of DNA and RNA extraction from archival formalin-fixed tissue. AB - Archival, formalin-fixed, paraffin-embedded tissue is an invaluable resource for molecular genetic studies but the extraction of high quality nucleic acid may be problematic. We have optimised DNA extraction by comparing 10 protocols, including a commercially available kit and a novel method that utilises a thermal cycler. The thermal cycler and Chelex-100 extraction method yielded DNA capable of amplification by PCR from every block and 61% of sections versus 54% using microwave and Chelex-100, 15% with classical xylene-based extraction and 60% of sections using the kit. Successful RNA extraction was observed, by beta-actin amplification, in 83.7% sections for samples treated by the thermal cycler and Chelex-100 method. Thermal cycler and Chelex-100 extraction of nucleic acid is reliable, quick and inexpensive. PMID- 10454650 TI - Flexible working in family planning and reproductive health care. PMID- 10454651 TI - Contraceptive efficacy of the personal hormone monitoring system Persona. PMID- 10454652 TI - Prospective multicentre study comparing levonorgestrel implants with a combined contraceptive pill: final results. AB - Norplant is a hormonal, long term method of contraception requiring the sub dermal placement of six flexible capsules containing levonorgestrel in the inner aspect of the upper non-dominant arm. This open, prospective, multicentre, parallel group study, comparing the acceptability of Norplant and a combined pill, was originally designed to follow 700 subjects for five years, but was discontinued early. The main outcome criteria were duration of use and reason for discontinuation if appropriate. This final analysis, concerning the 364 subjects who used the implant and 307 subjects who used a gestodene/ethinyloestradiol combined pill, shows a statistically significantly higher (p<0.001) continuation rate amongst the implant users at 83.4 per cent compared to the pill users at 64.4 per cent at one year and this difference decreased only slightly by two years. The most common reasons for discontinuation by the end of the study were adverse events in both groups (menstrual and then mood changes in the implant group and mood changes and then headaches in the pill group). From the whole cohort the proportion which were 'very satisfied' were higher in the implant group (28.5 per cent compared to 14.6 per cent). This study confirms, in the UK, a high continuation rate for implant users similar to other countries as reported in the literature. If acceptability and continuation rates are similar in the none study setting, then a substantial number of women will seek removal of the implant and possible reinsertion five years after the time of initial high popularity (ie 1999). Doctors need to be prepared for this possible high level of activity. PMID- 10454653 TI - Dual provision or duplication? A survey of family planning provision. AB - BACKGROUND: Family planning has been delivered through dual provision by general practice and community based clinics since its inception. This may be perceived as duplication of services and can be regarded as an area of possible disinvestment in a climate of rising health care costs. AIM: A survey of family planning service provision across a health district was carried out to establish the potential to rationalise current service provision by studying the pattern of service provision in the district and the links between family planning clinics and general practices. METHOD: A postal questionnaire and covering letter were sent to all practices and clinics in May 1997. Following analysis, results were,fed back to clinic staff and general practitioners through accredited meetings. RESULTS: Basic family planning was universally available to the population at all practices and clinics. Clinics offered a wider range of services, especially specialist services for psychosexual counselling and hormonal implants. Very few separate services for young people were identified. Within the primary care sector there was little 'out-of-hours' provision of contraceptive services. Half of the practices responding had used or would use family planning services as referral centres; implants and psychosexual services attracted most referrals. CONCLUSION: An understanding of the complementary nature of the services in primary care and community FP clinics was achieved and agreement was reached that disinvestment in clinics locally was not appropriate. The need to raise public awareness of availability of all contraceptive services was identified. Services in general practice and community clinics are complementary and need to develop a joint strategy to ensure an effective, comprehensive service. Quality of care needs to be examined in future work. PMID- 10454654 TI - Safer sex at holiday centres: providing contraceptive services to seasonal workers. AB - Many seasonal workers experience an increase in sexual activity whilst employed at a holiday centre. Evidence of sexual risk-taking while at a holiday centre has important social and health implications for purchasers and providers of sexual health services in areas which experience an annual influx of seasonal workers. This research investigates the contraceptive behaviour of seasonal workers and focuses on their access to contraception and sexual health services. In-depth interviews were conducted with seasonal workers at holiday centres along the south coast of England. Respondents were, interviewed at the beginning of the season and again, five months later at the end of the season. This longitudinal methodology enabled changes in contraceptive behaviour to be identified as well as the strategies for seeking contraception and sexual health services throughout the season. The results of this study show that there are a range of different motivations which influence seasonal worker' use of contraception and sexual risk taking while at a holiday centre. Categories of contraceptive protection are developed to assist purchasers and providers to identify the variety of sexual health needs of workers at holiday centres and determine the most effective strategies for delivering contraceptive and sexual health services to these workers. The paper describes the motivations which influence contraceptive use and sexual risk-taking amongst seasonal workers, identifies the contraceptive and sexual health needs of these workers, and discusses the difficulties workers experienced in meeting these needs while at a holiday centre. PMID- 10454655 TI - Changes in the sexual lifestyles of young people in Somerset, 1990-1996. AB - OBJECTIVES: The immediate objective was to up-date knowledge of the socio-sexual lifestyles and AIDS awareness of young people (16-24 years of age) in Somerset, and to compare these 1996 survey findings to the Somerset Survey findings from 1990. The ultimate objective was to provide information to assist the continuing development of HIV risk-reduction and sexual health promotion strategies for young people. DESIGN: An identical schedule-structured survey strategy to that employed in 1990 was undertaken in May to July 1996 among 498 young people throughout Somerset. A quota sampling strategy was undertaken in 50 electoral wards, randomly selected in proportion to population size. The interviews, undertaken in respondents' homes, involved a combined personal interview and self completion booklet. RESULTS: Between 1990 and 1996 broad shifts in sexual lifestyles were identified, involving earlier ages of first intercourse, slight increases in numbers of sexual partners and more 'casual-recreational' sexual attitudes. These changes were accompanied by a substantial increase in condom use. CONCLUSIONS: The trend among youth in Somerset is towards more open sexual lifestyles, but with the positive public health implications of higher levels of 'safer sex' practice. PMID- 10454656 TI - Knowledge of emergency contraception amongst female patients attending a department of genitourinary medicine. AB - The aim of the study was to assess the knowledge of emergency contraception amongst new female patients attending an inner-city department of genitourinary medicine. Information was also sought about use of regular contraception and demography. Three hundred and ninety nine questionnaires were suitable for analysis. Half of the sample answered that the latest a woman could take emergency contraception after unprotected sex was three days. None of the sample knew that emergency contraception could be obtained up to five days. Twenty nine per cent of the sample reported sex without contraception during the menstrual cycle preceding attendance. Women who had ever used regular contraception in the past were statistically less likely to have reported unprotected sex in the menstrual cycle preceding attendance (p=0.0000068). Professional women were statistically less likely to have reported unprotected sex in the menstrual cycle preceding the clinic visit. Fourteen per cent of the sample had genital warts at this first clinic visit, 10 per cent had Chlamydia trachomatis, seven per cent had herpes simplex infection, six per cent had gonococcal infection and five per cent had trichomonal infection. Women who reported unprotected sex during the preceding menstrual cycle were not statistically more likely to have a sexually transmitted infection at this first clinic visit. A large number of women attending departments of genitourinary medicine are at risk of both pregnancy and also sexually transmitted infection. Staff working in all areas of sexual health need to have a good knowledge of both contraception and sexually transmitted infections in order to educate the clients on both aspects of unprotected sex. PMID- 10454657 TI - Barriers to the use of IUDs as emergency contraception. AB - The intrauterine contraceptive device (IUD) is a very effective form of emergency contraception (EC). This author hypothesised that IUDs are an underused method and determined to evaluate potential barriers to IUD use. A postal survey of 100 family planning doctors and 100 general practitioners was conducted in Trent Region during March 1998 with a 70 per cent response rate. Lack of time was the most important factor that influenced doctor's decisions not to offer IUDs to the majority of women requesting emergency contraception. Most doctors registered concern about the risk of pelvic inflammatory disease. Misconceptions and a lack of accurate information contributed to participants reluctance to discuss IUDs as emergency contraception. Lack of time in consultations is a well-recognised issue in general practice. The risk of sexually transmitted infections is a nationwide concern, but is difficult to address without accurate data on the prevalence of the most common pathogens. Considerable effort would be required to increase doctors' knowledge and willingness to offer IUDs routinely to women requesting emergency contraception. PMID- 10454659 TI - Education and training module for a specialist registrar: a move forward in specialist registrar education. AB - A new post was created in Portsmouth. This produced the opportunity, for a new and innovative approach to the education and training module for the specialist registrar (SpR) training programme. The GP tutor with supervision from the university, designed, implemented and evaluated the module. It took place during the first five months of the SpR contract. The method used was a case study in which the SpR was encouraged to reflect on practice. The learning outcomes were documented initially by the GP tutor and,finally, by the SpR using a logbook based on the dental professional development log. Reflecting on practice enabled the SpR to produce the evidence of her own learning and plan her future learning needs. The project highlights the advantages and problems of reflecting on practice. These issues must be addressed if the conflict between the need to produce the competent doctor and the demands of an educationally sound programme are to be resolved, to create critical thinking and autonomous lifelong, self directed learners. PMID- 10454658 TI - Depo Provera. Position paper on clinical use, effectiveness and side effects. AB - Depo Provera (medroxyprogesterone acetate, DMPA) when given as 150 mg by deep intramuscular injection every 12 calendar weeks (84 days+5 days), is a highly effective contraceptive with a very low failure rate comparable to modern copper IUDs and lower than many other methods. It should be available as a first line method to all who wish to make an informed choice about reversible methods of contraception. Pre-use counselling is essential to minimise the effect of menstrual change which occurs in most patients. However there is great patient variability. Use of DMPA is independent of intercourse and also independent of the user's memory (and thus of continuing motivation), other than remembering the 12 weekly appointments. For many women this is a great advantage. Oral contraceptive methods involve remembering to take a pill each day, in the case of the progestogen only pill within the same three hours each day. This places considerable strain on women who lead irregular lifestyles, who are very busy or travel frequently. Such women often describe a constant 'fear of forgetting', especially with the POP. The main potential disadvantage of DMPA in this country are likely to be menstrual disturbance and weight gain. The combined oral contraceptive pill gives the appearance of excellent cycle control because it removes the natural cycle altogether and replaces it with an artificial one. All progestogen-only methods, whether low or high dose, lead to menstrual disturbances, so in this respect DMPA is not unique. Although troublesome, the menstrual disturbances which occur in DMPA users very rarely require operative medical intervention, and can often be improved simply by short courses of oestrogen or shorter injection intervals. Again, women need to know what can be done so that they are aware that they should seek advice early, rather than miserably waiting.for their 12 week appointment. DMPA has no appreciable effects on blood pressure or thrombosis risk. In this it has an advantage over the combined oral contraceptive pill, and provides a simple, effective alternative for women who cannot use the pill for these reasons. Similarly, it has been suggested that women who suffer from focal migraine and are therefore advised against use of the combined oral contraceptive pill can still use progestogen only contraceptives. Although the POP is medically safe in these circumstances, in young women it is less effective, and involves strict time keeping, which will be disadvantageous for some women. Side effects, long term use and schedules of administration are also discussed. The use of local protocols to allow nurse administration is to be supported both in general practice and the clinic situation. Perhaps the most important issue surrounding the use of DMPA is that of patient information. The method has had a particularly bad public image, which naturally makes potential users anxious and subject to misinformation from poorly informed or biased sources. Also, it is temporarily irreversible during its three months duration, so the duration of any problems or anxieties resulting from side effects may be longer than for other methods. It is of paramount importance that easily understood, accurate patient information leaflets are available, since biased and inaccurate information is readily available from women's magazines, perpetuating the myths surrounding the method. PMID- 10454660 TI - Where did family planning come from and where is the specialty going? PMID- 10454661 TI - Postcoital vaginal tear. PMID- 10454662 TI - Images, imagination and ideas: a perspective on the impact of ultrasonography on the practice of obstetrics and gynecology. Presidential address. PMID- 10454663 TI - Cervical incompetence: elective, emergent, or urgent cerclage. AB - OBJECTIVE: We define criteria for insertion of cervical cerclage done electively (historical), urgently (in patients without symptoms), or emergently (in patients with symptoms). We compare outcomes as determined by prolongation of pregnancy and survival in each of these groups, and we define whether urgent or emergent cerclage imparted a higher risk of spontaneous rupture of membranes or chorioamnionitis than that associated with elective cerclage. STUDY DESIGN: This is a retrospective analysis of all cervical cerclages placed at Memorial Medical Center between January 1, 1993, and December 31, 1997. Outcomes oberved were as follows: (1) prolongation of pregnancy in weeks after cervical cerclage, (2) presence of spontaneous rupture of membranes or clinical chorioamnionitis necessitating delivery, and (3) neonatal outcome. RESULTS: For prolongation of pregnancy the following results were obtained: emergent cerclage, 8.3 +/- 0.9 weeks; urgent cerclage, 12.2 +/- 1.5 weeks; and elective cerclage, 20.2 +/- 0.9 weeks (elective versus emergent and urgent, P <.05). For average gestational age at delivery, the results were as follows: emergent cerclage, 30.5 +/- 0.9 weeks; urgent cerclage, 33.1 +/- 1.4 weeks; and elective cerclage, 35.5 +/- 0.9 weeks (elective versus emergent and urgent, P <.05). The total neonatal survival was 85.7%. The incidence of spontaneous rupture of membranes was as follows: emergent cerclage, 51%; urgent cerclage, 40%; elective cerclage, 18% (elective versus emergent and urgent, P <.05). The incidence of clinical chorioamnionitis showed similar results. CONCLUSIONS: It is clear that emergency cerclages confer some benefit in patients with evidence of cervical incompetence. From this study it is evident that there is a new group of patients who need cerclage on an urgent basis as shown by subtle ultrasonographic changes in the cervix. Their behavior mirrors that of those belonging to the emergent group, suggesting that if they were left untreated they would need cerclage on an emergency basis. PMID- 10454665 TI - Aggressive perinatal care for high-order multiple gestations: Does good perinatal outcome justify aggressive assisted reproductive techniques? AB - OBJECTIVE: The purpose of this study was to determine the factors that must be considered for appropriate counseling of patients with high-order multiple gestations. STUDY DESIGN: A retrospective chart review was carried out from all high-order multiple gestations that were managed by a single perinatology group from February 1993-June 1998. Twin pregnancies that did not result from fetal reduction procedures were used as a control group. RESULTS: Clinical outcome data were analyzed from 9 quadruplet, 25 triplet, 19 reduced twin, and 24 nonreduced twin pregnancies. Women with quadruplet pregnancies were admitted more frequently at an early gestational age, the infants were delivered earlier, and the maternal and neonatal hospital days were longer than for triplet and reduced and nonreduced twin gestations. Triplet pregnancies had an earlier gestational age at delivery (32.3 vs 34.2 weeks), a higher incidence of preterm labor (87% vs 68%), and a higher percentage of neonatal intensive care unit admissions (94% vs 59%) than reduced twin gestations. Reduced twins were hospitalized longer (16.4 vs 9.8 days), were delivered earlier (34.2 vs 36.2 weeks), had a higher incidence of preterm labor (68% vs 29%), and had a greater percentage of neonatal intensive care unit admissions (59% vs 21%), a greater percentage of birth weight <1500 g, and a greater frequency of respiratory distress syndrome (16% vs 2%) than nonreduced twins. There was no difference in neonatal survival and neurologic morbidity when all groups were compared. CONCLUSION: Although early delivery and prolonged (maternal and neonatal) hospitalization were common with quadruplets and triplets, maternal and neonatal outcomes were excellent. The decision for reduction from triplets to twins may not necessarily change pregnancy outcome but should still be discussed as an option for the parents. Continued efforts need to be made to reduce the overall number of iatrogenic high-order multiple gestations. PMID- 10454664 TI - Singleton term breech deliveries in nulliparous and multiparous women: a 5-year experience at the University of Miami/Jackson Memorial Hospital. AB - OBJECTIVE: The purpose of this retrospective study was to evaluate the feasibility of planned vaginal delivery, the maternal morbidity and mortality, and the short-term perinatal outcome in selected multiethnic women at term with singleton breech presentations. STUDY DESIGN: Singleton breech deliveries were identified from the delivery database between January 1, 1989, and December 31, 1993. A retrospective chart review identified 310 nulliparous and 711 multiparous women at term (37-42 weeks) for a total of 1021. Parameters studied included the success rate of planned vaginal deliveries and the incidences of maternal morbidity, perinatal morbidity, and mortality as a whole stratified by parity and mode of delivery. The Student t test, chi(2) test, and Fisher exact test were used for statistical analysis. RESULTS: Among 1021 women with singleton fetuses in a breech position at term, 191 were candidates for vaginal delivery, and 135 (70.7%) of these deliveries were successful. By parity, 12.3% of 310 nulliparous women and 21.5% of 711 multiparous women were candidates for vaginal delivery; 50% of the former and 75.8% of the latter underwent vaginal delivery. Maternal morbidity was more commonly associated with multiparity and cesarean delivery. Newborn intensive care admissions were equally distributed by parity, and significantly more were for vaginal than cesarean deliveries (17.4% vs 10.8%, P =.036). Premature rupture of the membranes complicated deliveries in 23.9% of the nulliparous women and only 6.5% of the multiparous women (P =.000). CONCLUSION: In this multiethnic population 70.7% of candidates selected for attempted vaginal breech delivery at term were successful. The remaining 29.3% underwent cesarean delivery for labor disorders or nonreassuring fetal heart rate patterns. PMID- 10454666 TI - Choroid plexus cysts: Is biochemical testing a valuable adjunct to targeted ultrasonography? AB - OBJECTIVE: We sought to determine whether biochemical testing is a valuable adjunct to ultrasonography in selecting patients with fetal choroid plexus cysts for amniocentesis. STUDY DESIGN: The study population consists of 128 patients who had fetal choroid plexus cysts detected during ultrasonography performed between 18 and 22 weeks' gestation. The patients had genetic counseling, and amniocentesis and biochemical testing were offered to all patients. The data were analyzed by dividing the patients into 3 groups. Group 1 had targeted ultrasonography only, group 2 had ultrasonography and maternal serum alpha fetoprotein testing, and group 3 had ultrasonography and triple-screen (maternal serum alpha-fetoprotein, human chorionic gonadotropin, and estriol) testing. Outcome was determined by fetal karyotype or by neonatal examination by a pediatrician for patients who declined amniocentesis. RESULTS: There were 25 patients in group 1. Isolated choroid plexus cysts were detected in 20 fetuses, and all had normal outcomes. Additional anomalies were detected in 5 fetuses. Two had normal karyotypes, and 3 had trisomy 18. There were 52 patients in group 2. The maternal serum alpha-fetoprotein levels were normal in 44 patients, 41 of whom had isolated fetal choroid plexus cysts. Of these 44 patients, 40 had normal outcomes, and 1 patient had a fetus with trisomy 18. The remaining 3 patients with normal maternal serum alpha-fetoprotein levels had additional fetal anomalies on ultrasonography, but the karyotypes were normal. The maternal serum alpha-fetoprotein levels were abnormal in 8 patients, of whom 6 had fetuses with isolated choroid plexus cysts and normal karyotypes. Two patients had additional fetal anomalies detected on ultrasonography and had abnormal karyotypes, 1 with trisomy 21 and 1 with trisomy 18. There were 51 patients in group 3. Results of the triple screen were normal in 32 patients. The choroid plexus cysts were isolated in 29 of the 32 patients, and all 29 fetuses had normal karyotypes. The other 3 patients with normal triple-screen results had additional fetal anomalies on ultrasonography. One fetus had normal chromosomes, and 2 had trisomy 18. The remaining 19 patients had abnormal triple-screen results. Among them, 16 fetuses had isolated choroid plexus cysts, 13 of whom were normal, 2 had trisomy 18, and 2 had a de novo unbalanced translocation. The remaining 3 fetuses had additional anomalies, and all 3 fetuses had trisomy 18. There were 14 fetuses with significant chromosomal abnormalities. Nine mothers were <35 years old, and 5 were >/=35 years old. CONCLUSIONS: This study shows the following: (1) The triple screen is a useful adjunct to targeted ultrasonography in selecting patients with fetal choroid plexus cysts for amniocentesis. (2) A normal triple-screen result and the absence of additional fetal anomalies on ultrasonography reliably exclude an underlying chromosomal abnormality, and amniocentesis is not indicated. (3) If the triple-screen result is abnormal, additional anomalies are seen on ultrasonography, or the mother is aged >/=35 years, then fetal karyotyping is recommended. (4) Patients who decline fetal karyotyping should have follow-up ultrasonography in 34 weeks' time. PMID- 10454668 TI - Injury of the rectum during vaginal surgery. AB - OBJECTIVE: We sought to report our experience with injury of the rectum during vaginal surgery. STUDY DESIGN: A retrospective chart review of a vaginal surgery database was performed. RESULTS: Nine injuries were identified. Four patients with posthysterectomy vault prolapse incurred a rectal laceration just above the perineum during an attempt to develop the rectovaginal space. Another rectal laceration occurred just above the perineum during an attempt to develop the rectovaginal space during vaginectomy-hysterectomy for gender dysphoria. Three other injuries occurred between the vaginal cuff and peritoneum while posterior culdotomy for vaginal hysterectomy was attempted. The remaining injury was in a Schuchardt incision. All 9 injuries were recognized, repaired, and healed without complication. CONCLUSIONS: Laceration of the rectum is an uncommon complication of vaginal surgery. Prompt recognition and repair result in uncomplicated primary healing. PMID- 10454667 TI - Urinary incontinence in pregnancy and the puerperium: a prospective study. AB - OBJECTIVE: Pregnancy and childbirth are commonly thought to be associated with the development of urinary incontinence and lower urinary tract symptoms. The purpose of this study was to assess the relationship, if any, between pregnancy and the development of lower urinary tract symptoms. STUDY DESIGN: A prospective study of lower urinary tract symptoms was carried out in a cohort of pregnant women who answered a series of symptom questionnaires and kept a 24-hour bladder chart on which frequency of urination and volumes voided were recorded throughout pregnancy and for 8 weeks after birth. RESULTS: A total of 123 women participated in the study. Mean daily urine output (P =.01) and the mean number of voids per day (P =.01) increased with gestational age and declined after delivery. Episodes of urinary incontinence peaked in the third trimester and improved after birth (P =.001). White women had higher mean voided volumes and fewer voiding episodes than did black women. Ingestion of caffeine was associated with smaller voided volumes and greater frequency of urination. CONCLUSION: Pregnancy is associated with an increase in urinary incontinence. This phenomenon decreases in the puerperium. Pregnancy and childbirth trauma are important factors in the development of urinary incontinence among women. These findings warrant further investigation. PMID- 10454669 TI - Spontaneous resolution rate of grade 1 cervical intraepithelial neoplasia in a private practice population. AB - OBJECTIVE: The purpose of this study was to assess the rate of spontaneous resolution of grade 1 cervical intraepithelial neoplasia in a private practice patient population. STUDY DESIGN: One hundred consecutive women with biopsy proven grade 1 cervical intraepithelial neoplasia were offered expectant management in the form of repeat Papanicolaou smears every 3 months up to 1 year or treatment. Women with inadequate colposcopic examination, positive endocervical sampling, or discordant Papanicolaou smears were excluded. RESULTS: Eighty-nine (89%) women agreed to undergo expectant management. Of these, 67 (75.3%) experienced spontaneous resolution of lesions on the basis of 2 subsequent and consecutive normal smears corroborated by colposcopy. The median time to resolution was 9 months (n = 38). Seventeen (19.9%) women were treated at 1 year for persistent atypical squamous cells of undetermined significance or low grade squamous intraepithelial lesions by loop electrosurgical conization. Of these, 1 (1.1%) had a more advanced lesion (grade 2 cervical intraepithelial neoplasia). Four (4.5%) had progression before 1 year and were treated. Only 1 (1.1%) patient was lost to follow-up. CONCLUSION: Expectant management of grade 1 cervical intraepithelial neoplasia is safe and cost-effective in a reliable patient population. PMID- 10454670 TI - Cervical wet mount as a negative predictor for gonococci- and Chlamydia trachomatis-induced cervicitis in a gravid population. AB - OBJECTIVE: The purpose of this study was to determine whether a wet mount preparation of endocervical secretions from obstetric patients can accurately rule out the presence of gonococci and Chlamydia trachomatis. STUDY DESIGN: Gravid patients were screened for gonococci and C trachomatis with a deoxyribonucleic acid probe and a wet mount preparation of endocervical secretions. The number of polymorphonuclear lymphocytes was counted and averaged per x400 high-power field. A count of <10 polymorphonuclear lymphocytes per high power field was defined as predicting the absence of gonococci- and C trachomatis -induced cervicitis. A chi(2) analysis was used to compare the wet mount results to the deoxyribonucleic acid probe. RESULTS: Between January and March 1998, 341 patients were enrolled in the study. Wet mount results showed 32% (108/341) of patients had <10 polymorphonuclear lymphocytes per high-power field. The incidence of infection with gonococci and C trachomatis in this population, as determined by deoxyribonucleic acid probe, was 10 of 341 (2.9%) and 30 of 340 (8.8%), respectively. The sensitivity values for gonococci and C trachomatis were 90% and 87%, respectively. The negative predictive value of the wet mount for gonococci-induced cervicitis was 99%, and that for C trachomatis -induced cervicitis was 96%. CONCLUSION: In this population the wet mount accurately predicted the absence of gonococci- and C trachomatis -induced cervicitis. Use of this screening protocol in gravid patients has the potential to reduce the cost of screening for these 2 diseases. PMID- 10454671 TI - Estrogen replacement therapy in women with previous breast cancer. AB - OBJECTIVE: We sought to review the status of patients with breast cancer who were treated with estrogen replacement therapy and compare the results with those of nonestrogenic hormone users and women not treated with hormone replacement. STUDY DESIGN: The study group consisted of 76 patients with breast cancer, including 50 using estrogen replacement for up to 32 years, 8 using nonestrogenic hormone replacement for up to 6 years and followed for up to 11 years, and 18 using no hormones for up to 10 years. In addition to estrogen use, 40 of the 50 hormone users were treated with androgens, usually in the form of implantation of testosterone pellets. Forty-five subjects were also given progestogens, usually megestrol acetate 20 to 40 mg for 10 to 25 days each month. The 8 nonestrogen hormone users were treated with various combinations of testosterone pellets, tamoxifen, and progestogens. Forty-two of the 50 estrogen users are still being treated in our clinic, as are 2 of the 8 subjects using nonestrogen hormone. Follow-up was done through the tumor registry at University Hospital, and those whose tumor records were not current were telephoned. RESULTS: Of the 50 estrogen users, 3 have died (a mortality rate of 6%), and the rest have been followed for 6 months to 32 years, with a mean duration of follow-up of 83.3 +/- 8.81 months. One of the 8 nonestrogen hormone users has died (a mortality rate of 12.5%), and the rest have been followed for 2 to 11 years, with a mean duration of follow-up of 72.0 +/- 5. 93 months. Six of the 18 women not using hormone replacement have died (a mortality rate of 33.3%), and the rest have been followed for 6 months to 10 years, with a mean duration of follow-up of 50.5 +/- 6.01 months. CONCLUSION: Estrogen replacement therapy apparently does not increase either recurrences or mortality rates. Adding progestogens may even decrease recurrences. Women with early breast cancer should be offered hormone replacement therapy after a full explanation of the benefits, risks, and controversies. PMID- 10454672 TI - Cerebral arteriovenous malformation in pregnancy: presentation and neurologic, obstetric, and ethical significance. AB - Cerebral arteriovenous malformations infrequently complicate pregnancy. We sought to determine the neurologic, obstetric, and ethical significance of such malformations. We present the clinical course of 2 pregnant women with arteriovenous malformations who experienced cerebral hemorrhage and a loss of capacity for decision making. We also review the neurologic and obstetric significance of arteriovenous malformations in pregnancy. Various treatment options with concern for pregnancy and the prognosis for arteriovenous malformations are outlined. The ethical issues involved for pregnant patients whose decisional capacity is compromised as a result of cerebral injury are explored. A review of persistent vegetative state and brain death (death by neurologic criteria) occurring in pregnancy allows us to explore many issues that are applicable to decisionally incapacitated but physiologically functioning pregnant women. We outline a document, the purpose of which is to obtain advance directives from pregnant women regarding end-of-life decisions and to appoint a surrogate decision maker. We believe that evaluation and treatment of the arteriovenous malformation may be undertaken without regard for the pregnancy and that the pregnancy should progress without concern for the arteriovenous malformation. PMID- 10454673 TI - HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome: the benefit of corticosteroids. AB - OBJECTIVE: The purpose of this study was to determine the effect of corticosteroids on platelet counts and liver functions in women with pregnancies complicated by the HELLP (hemolysis, elevated liver enzymes, and low platelets) syndrome. STUDY DESIGN: The study group consisted of 93 patients between 24 and 34 weeks' gestation diagnosed with HELLP syndrome. All were given intramuscular injections of either betamethasone or dexamethasone. The 3 most common regimens used were 12 mg of intramuscular betamethasone administered twice 12 hours apart, 12 mg of intramuscular betamethasone administered twice 24 hours apart, and 6 mg of intramuscular dexamethasone administered 4 times 6 hours apart. Precorticosteroid and postcorticosteroid platelet counts and liver function test results were compared. The differences in improvement in hematologic abnormalities among the 3 corticosteroid regimens were also analyzed. RESULTS: The hematologic abnormalities seen in the 93 patients with HELLP syndrome improved after the administration of corticosteroids. The platelet count increased by 23.3 x 10(3)/microL (P <.001). A statistically significant decrease was seen in liver enzyme levels. The alanine aminotransferase decreased by 31.6 IU/L, the aspartate aminotransferase decreased by 52.1 IU/L, and the alkaline phosphatase decreased by 7.6 IU/L. Of the 3 regimens used, 2 doses of 12 mg of intramuscular betamethasone given every 12 hours improved the liver function to the greatest degree. CONCLUSIONS: This study demonstrates that corticosteroids produce a significant improvement in the hematologic abnormalities associated with HELLP syndrome. Two doses of betamethasone given 12 hours apart was the most effective corticosteroid regimen. PMID- 10454674 TI - Change in antibiotic resistance of group B streptococcus: impact on intrapartum management. AB - OBJECTIVE: Intrapartum chemoprophylaxis has resulted in a significant reduction of group B Streptococcus neonatal infection. For penicillin-allergic patients, clindamycin or erythromycin is the recommended antibiotic. The purpose of this study was to establish any pattern of antibiotic resistance of group B streptococcal clinical isolates over the past 15 years. STUDY DESIGN: Group B streptococcal isolates obtained from the lower genital tract were tested for sensitivity to ampicillin, penicillin, clindamycin, and erythromycin. The sensitivity of 100 group B streptococcal isolates retrieved in the period 1997 1998 was compared with that of 85 group B streptococcal isolates from 1980-1993. RESULTS: From 1980-1993 group B streptococcal isolates were available for testing for antibiotic resistance along with 100 isolates from a second study period 1997 1998. Of the 100 group B streptococcal isolates from 1997-1998, 18 were resistant to erythromycin, of which 5 were also resistant to clindamycin, as compared with 1 of the 85 isolates from 1980-1993 that was resistant to erythromycin (P <.001). All the isolates were sensitive to ampicillin and penicillin. All 18 resistant strains from 1997-1998 were found to be sensitive to cephalothin. CONCLUSION: Over the past 18 years there has been increased in vitro resistance of group B streptococci to both clindamycin and erythromycin. If other studies confirm these findings, modifications to the current Centers for Disease Control and Prevention recommendations may be necessary. PMID- 10454675 TI - Peripartum cardiomyopathy: summary of an international survey on peripartum cardiomyopathy. AB - OBJECTIVE: The aim of this survey was to assess the evaluation, management, and future recommendations of patients with the diagnosis of peripartum cardiomyopathy and to evaluate the interest in the creation of a prospective database regarding this rare disorder. STUDY DESIGN: A total of 116 surveys were sent to major teaching institutions in the United States (including Puerto Rico), Canada, Crete, and South Korea after a national conference held at the National Institutes of Health regarding peripartum cardiomyopathy. This was an open-ended survey containing 17 specific questions regarding this disorder and its management. RESULTS: A total of 78 (67%) maternal-fetal specialists responded to the survey. Diuretics and digoxin were used as first-line treatment for this disorder. Only 6% used angiotensin-converting enzyme inhibitors during pregnancy. Fifty-eight percent of the perinatologists (58%) recommended either intrauterine contraceptive devices or foam and condoms, whereas oral contraceptives (progesterone-only pill or estrogen-progesterone mix) were recommended in 23% and 41%, respectively. Sixty-six percent (66%) of the respondents would recommend future pregnancy if ventricular function returned to normal. CONCLUSIONS: Fundamental clinical and basic research is lacking regarding this rare but potentially devastating disorder. Major teaching institutions do not have significant numbers of patients with this disorder to provide concrete recommendations, and starting a database will be useful in the gathering of important epidemiologic information. A secondary aim of such a registry would be to establish a repository for tissue and blood samples to answer basic mechanistic questions about this disorder. PMID- 10454676 TI - Neonatal sepsis after betamethasone administration to patients with preterm premature rupture of membranes. AB - OBJECTIVE: We sought to determine the effect of antenatal betamethasone exposure on the incidence of early onset neonatal sepsis in patients with preterm premature rupture of membranes. STUDY DESIGN: We performed a nonconcurrent prospective analysis of infants delivered between 24 and 34 weeks' gestation after preterm premature rupture of membranes. Patients with preterm premature rupture of membranes were categorized into 3 groups on the basis of the following betamethasone exposures: (1) none (control subjects), (2) two 12-mg doses in a 24 hour interval on admission (single course), and (3) weekly administration after the initial single course (multiple courses). All included patients received prophylactic antibiotics for group B streptococci. Discrete data were tested for significance with the chi(2) test. Continuous data were tested for significance with an analysis of variance. Multiple logistic regression analysis was performed to determine the confounding effect of the multiple variables that were considered risk factors for early-onset neonatal sepsis. All P values of <.05 were considered significant. RESULTS: Three hundred seventy-four patients with preterm premature rupture of membranes were included, 203 of whom were evaluated in the control group, 99 in the single-course group, and 72 in the group receiving multiple courses of betamethasone. Early-onset neonatal sepsis was significantly associated with multiple courses of corticosteroids (P <.001) and gestational age (P =.002). Multiple courses of antenatal betamethasone were significantly associated with chorioamnionitis (P =.004) and endometritis (P =.004). Single-course corticosteroid administration was not significantly associated with any maternal or neonatal infectious complications. CONCLUSIONS: Multiple courses of antenatal betamethasone administered to patients with preterm premature rupture of membranes is associated with an increased risk of early onset neonatal sepsis development. PMID- 10454677 TI - Managed care in obstetrics and gynecology: can our specialty survive? PMID- 10454678 TI - Prophylactic cesarean delivery for fetal macrosomia diagnosed by means of ultrasonography--A Faustian bargain? AB - Both our previously performed decision analysis and more recent clinical data considered in the context of our decision analytic framework support the claim that in the pregnancies of women without diabetes the level of intervention and the economic costs of prophylactic cesarean delivery for fetal macrosomia diagnosed by means of ultrasonography are predicted to be excessive. Under the most plausible assumptions, a prophylactic cesarean policy with either a 4000- or 4500-g macrosomia threshold would require more than 1000 cesarean deliveries and millions of dollars to avert a single permanent brachial plexus injury. In the pregnancies of diabetic women, although such policies would be expected to perform appreciably better, their use would nevertheless entail considerable intervention for any benefit achieved. Under most assumptions, hundreds of cesarean deliveries and hundreds of thousands of dollars would be required to avert a single permanent brachial plexus injury. In light of the available data, optimizing the management of shoulder dystocia seems at present to be the most immediate and tenable approach to the prevention of birth-related brachial plexus injury. PMID- 10454679 TI - Labor induction with misoprostol. AB - Misoprostol, a prostaglandin E(1 ) analog, is widely used in the United States for cervical ripening and labor induction. Its use for these indications is not approved by the US Food and Drug Administration. The only Food and Drug Administration-approved indication in the product labeling is the treatment and prevention of intestinal ulcer disease resulting from nonsteroidal anti inflammatory use. Multiple trials have proved that misoprostol is an effective agent for cervical ripening and labor induction in term pregnancy; however, investigations continue regarding the optimal dose, dosing regimen, and route of administration. Uterine contraction abnormalities are often found in association with higher misoprostol doses. Some trials also indicate increased frequencies of meconium passage, neonatal acidemia, and cesarean delivery for fetal distress in women receiving higher doses of misoprostol. Overall, most trials fail to demonstrate a significant change in the cesarean delivery rate with the use of this agent. Misoprostol is an effective agent for cervical ripening and labor induction when used in a judicious and cautious fashion. PMID- 10454680 TI - Correlation of asbestos fiber burdens in fallopian tubes and ovarian tissue. AB - Evidence suggests an increased risk of ovarian cancer with asbestos exposure. Ovaries and corresponding fallopian tubes were studied by analytic electron microscopy. There was 71.4% agreement between tube and ovary for presence-type of asbestos. The fallopian tube can provide useful information regarding asbestos exposure when no ovarian tissue is available. PMID- 10454681 TI - Mammographic breast density during hormone replacement therapy: differences according to treatment. AB - OBJECTIVE: Our purpose was to investigate the effects of various hormone replacement regimens on mammographic breast density. STUDY DESIGN: Mammographic density was recorded in women participating in a population-based screening program. All women were nonusers of hormone replacement therapy at first mammogram and thereafter reported continuous use of the same treatment: estrogen alone (n = 50) or estrogen in cyclic (n = 75) or continuous (n = 50) combination with progestogen. Mammographic density was quantified according to the Wolfe classification. RESULTS: An increase in mammographic density was much more common among women receiving continuous combination hormone replacement therapy (52%) than among those receiving cyclic (13%) and estrogen-only (18%) treatment. The increase in density was apparent already at first visit after the start of hormone replacement therapy. There was little change in mammographic status during long-term follow-up. CONCLUSION: Regimens of hormone replacement therapy were shown to have different effects on the normal breast. There is an urgent need to clarify the biologic nature and significance of a change in mammographic density during treatment and, in particular, its relation to symptoms and breast cancer risk. PMID- 10454682 TI - Surgical resection alone is effective treatment for ovarian immature teratoma in children and adolescents: a report of the pediatric oncology group and the children's cancer group. AB - OBJECTIVE: In both adult women and children the potential for malignant recurrence from ovarian immature teratoma has prompted the standard use of chemotherapy after complete resection of the primary tumor. The efficacy of postoperative chemotherapy in children and adolescents with ovarian immature teratoma, however, has not been established. A pediatric intergroup trial (INT 0106) was designed to determine the need for postoperative chemotherapy in patients with ovarian immature teratoma after management with surgical resection only. STUDY DESIGN: Between 1990 and 1995, 44 patients with completely resected ovarian immature tumor and without postoperative chemotherapy, who were able to undergo assessment, were accrued. Tumor tissue was evaluated by central pathology review to confirm diagnosis and determine tumor grading of immature neural elements. Patients were followed carefully for recurrence of disease with appropriate diagnostic imaging and serum marker studies. RESULTS: Thirty-one patients had pure ovarian immature teratoma with a tumor grade of 1 (n = 17), 2 (n = 12), or 3 (n = 2). Age at diagnosis ranged between 1.5 and 15 years (median, 10). Of the 29 patients studied, the serum alpha-fetoprotein level was elevated in 10 (34%); the median level was 25 ng/ml. Thirteen patients had ovarian immature teratoma plus microscopic foci of yolk sac tumor. Tumor grade was 1, 2, or 3 in 1, 6, and 6 patients, respectively. Age ranged between 6 and 20 years (median, 12). In the 12 patients evaluated for serum alpha-fetoprotein, 10 (83%) had elevated levels; the median level was 262 ng/ml. The 4-year event-free and overall survival for the ovarian immature teratoma group and for the ovarian immature teratoma plus yolk sac tumor group was 97.7% (95% confidence interval, 84.9%-99.7%) and 100%, respectively. The only yolk sac tumor relapse occurred in a child with ovarian immature teratoma and yolk sac tumor who was then treated with chemotherapy and is alive and free of disease 57 months after recurrence. CONCLUSION: The results of this study suggest that surgery alone is curative for most children and adolescents with resected ovarian immature teratoma of any grade, even when elevated levels of serum alpha-fetoprotein or microscopic foci of yolk sac tumor are present. This experience strongly supports avoiding the long-term effects of chemotherapy in most children with ovarian immature teratoma by reserving postoperative therapy for cases with relapse. PMID- 10454683 TI - The nuclear transcription factor NF-kappaB mediates interleukin-1beta-induced expression of cyclooxygenase-2 in human myometrial cells. AB - OBJECTIVE: Up-regulation of prostaglandin production by gestational tissues in the setting of intrauterine infection has been implicated as an important contributor to preterm labor and parturition. In this study we investigated the possible role of the nuclear transcription factor NF-kappaB in interleukin-1 signaling, leading to the expression of cyclooxygenase 2 and prostaglandin production in human myometrial cell cultures. STUDY DESIGN: Human myometrial smooth muscle cells from an immortalized line were used as a model system between passages 20 and 35. Growth-arrested cell cultures were stimulated with human recombinant interleukin 1, and the activation of NF-kappaB was assessed by the degradation of the inhibitory protein IkappaB-alpha (Western analysis), as well as by nuclear binding of NF-kappaB by using an electrophoretic mobility shift assay. The abundance of cyclooxygenase-2 messenger ribonucleic acid and protein was measured by Northern and Western analyses, whereas prostaglandin (prostaglandin I(2 ) and prostaglandin E(2 )) production was determined by specific radioimmunoassays. RESULTS: Within 15 minutes of stimulation with interleukin 1, 90% of IkappaB-alpha was degraded. This was temporally associated with nuclear translocation and binding of NF-kappaB. Within 30 minutes, cyclooxygenase 2 messenger ribonucleic acid appeared, with steady-state levels increasing up to 4 hours. This was followed by an up to 80-fold increase in cyclooxygenase 2 protein and a corresponding time-dependent increase in prostaglandin production. When IkappaB-alpha degradation was blocked with calpain I inhibitor, NF-kappaB translocation, cyclooxygenase 2 messenger ribonucleic acid and protein expression, and prostaglandin synthesis were also inhibited. CONCLUSION: Stimulation of human myometrial cells with interleukin 1 leads to rapid activation of the transcription factor NF-kappaB, which is functionally linked to the expression of cyclooxygenase 2 messenger ribonucleic acid, protein, and prostaglandin synthesis. PMID- 10454684 TI - Is the lack of concurrence of bacterial vaginosis and vaginal candidosis explained by the presence of bacterial amines? AB - OBJECTIVE AND STUDY DESIGN: We report for the first time an inhibitory effect on cell division and germ tube formation by Candida albicans and strains of other Candida species by putrescine and cadaverine. RESULTS: Both bacterial amines showed a dose-dependent inhibition of germ tube formation by C albicans, as well as budding (inhibition of cell division) of strains of other Candida species (ie, C glabrata, C krusei, and C tropicalis). CONCLUSIONS: We hypothesize that the presence of these and possibly other bacterial amines produced by anaerobes in the vaginal flora and seen in bacterial vaginosis, as in the healthy gut, may explain why candidosis is rarely seen in these instances. PMID- 10454685 TI - Management of long-standing unexplained infertility: A prospective study. AB - OBJECTIVE: Our purpose was to evaluate a protocol for the management of long standing unexplained infertility. STUDY DESIGN: A prospective study was conducted at an in vitro fertilization-embryo transfer center with 485 patients with the diagnosis of long-standing unexplained infertility. Patients were treated by means of controlled ovarian hyperstimulation and intrauterine insemination for a maximum of 3 cycles. Those who did not become pregnant underwent in vitro fertilization and intracytoplasmic sperm injection on sibling oocytes. RESULTS: Among 485 patients, 921 cycles of controlled ovarian hyperstimulation and intrauterine insemination were performed. The pregnancy rate was 15.7% per cycle and 29.8% per patient. Among those who did not become pregnant, 131 patients underwent in vitro fertilization and intracytoplasmic sperm injection into sibling oocytes; 48 women became pregnant (36.7%). There was total failure of the in vitro fertilization oocytes in 23 cycles (17.6%). CONCLUSION: Management of long-standing unexplained infertility with controlled ovarian hyperstimulation and intrauterine insemination resulted in a reasonable pregnancy rate. Treatment by means of in vitro fertilization and intracytoplasmic sperm injection into sibling oocytes among patients who did not become pregnant resulted in a good pregnancy rate. The overall results showed that the proposed protocol for management of unexplained infertility is satisfactory. PMID- 10454686 TI - Laparoscopic lymphadenectomy and vaginal or laparoscopic hysterectomy with bilateral salpingo-oophorectomy for endometrial cancer: morbidity and survival. AB - OBJECTIVES: Our goal was to evaluate the morbidity, recurrence, and survival of patients with clinical stage I endometrial cancer treated by laparoscopic lymphadenectomy with vaginal or laparoscopic hysterectomy and bilateral salpingo oophorectomy. STUDY DESIGN: This article is a retrospective review of records for 56 patients. The mean follow-up among those alive at last contact was 2.4 years (range, 32 days-5.2 years). Staging according to the International Federation of Gynecology and Obstetrics (1988) was as follows: I, 45 (80.4%); II, 3 (5.4%); III, 6 (10.7%); and IV, 2 (3.6%). RESULTS: Intraoperative complications occurred in 4 patients (7.1%). Transformation to laparotomy was necessary in 7 patients. Postoperative complications were observed in 9 patients (16.1%). Pelvic irradiation was administered postoperatively to 11 patients (19.6%). Among the 45 patients with surgical stage I disease, the 3-year recurrence rate was 2.5% and the 3-year cause-specific survival was 96.0%. CONCLUSIONS: Laparoscopic lymphadenectomy and vaginal or laparoscopic hysterectomy with bilateral salpingo oophorectomy provided 3-year survival and recurrence rates similar to those of the traditional abdominal approach. PMID- 10454687 TI - Assessing gains in diagnostic utility when human papillomavirus testing is used as an adjunct to papanicolaou smear in the triage of women with cervical cytologic abnormalities. AB - OBJECTIVE: We aimed to provide simple methods for calculating expected sensitivity and specificity when an adjunctive test is added to a conventional test. STUDY DESIGN: Use of adjunctive methods for the triage of women with cervical abnormalities produces an apparent gain in sensitivity over Papanicolaou cytologic testing alone. This increase in sensitivity can be misleading, even if deemed significant by results of a statistical test. Combined testing prevents a loss in specificity but sometimes offers no real gain in sensitivity. A nominal increase in sensitivity always occurs by chance whenever an adjunctive test is used in parallel with a conventional one, even if the new test is totally random with respect to the disease being evaluated. RESULTS: Gains in sensitivity and losses in specificity have to be gauged against expected levels of these parameters when a random adjunctive test is coupled with Papanicolaou screening and not gauged against the performance of cytologic testing alone. CONCLUSION: We provide simple formulas for calculating the expected sensitivity and specificity in conditions of combination testing to provide more realistic baselines for assessment of the screening efficacy contributed by the adjunctive test. PMID- 10454688 TI - Matrix metalloproteinase 9 activity in urine of patients at risk for premature delivery. AB - A simple, noninvasive assay was used to quantitate urinary matrix metalloproteinase 9 activity among 15 patients with threatened premature labor. Both positive and negative predictive values for risk for premature delivery were 80%. PMID- 10454689 TI - Reversible peripartum liver failure: a new perspective on the diagnosis, treatment, and cause of acute fatty liver of pregnancy, based on 28 consecutive cases. AB - OBJECTIVE: We sought to describe our experience with the clinical diagnosis, management, and course of patients with acute fatty liver of pregnancy. STUDY DESIGN: Twenty-eight cases of acute fatty liver of pregnancy at the Los Angeles County and University of Southern California Medical Center from 1982 to June 1997 were identified, and presenting symptoms, clinical course, laboratory values, maternal complications, and neonatal outcomes were studied. RESULTS: The incidence of acute fatty liver of pregnancy was 1 in 6659 births. There were no maternal deaths. Initial presentation was at an average of 37 weeks of gestation with a characteristic prodrome of malaise, nausea, vomiting, and abdominal pain. No patient was admitted with the diagnosis of acute fatty liver of pregnancy. The condition was diagnosed most commonly on the second hospital day after laboratory results indicated coagulopathy, renal insufficiency, and liver function abnormalities. One patient underwent liver biopsy at cesarean delivery. Radiologic studies did not aid with the diagnosis. Twenty-one patients were admitted in spontaneous labor, and 16 labors were complicated by abnormal fetal heart rate patterns or meconium. There was 1 stillbirth and 1 neonatal death as a result of perinatal asphyxia. Maternal morbidity consisted of hypoglycemia, infection, renal insufficiency, coagulopathy, encephalopathy, and wound complications. All patients had evidence of disseminated intravascular coagulopathy with profoundly decreased antithrombin levels. All patients recovered normal liver function post partum. CONCLUSIONS: Reversible peripartum liver failure may be diagnosed and managed on the basis of clinical and laboratory criteria. With adequate support, these patients may have full recovery of hepatic function. PMID- 10454690 TI - Epidemiology of miscarriage and its relation to other reproductive events in Finland. AB - OBJECTIVE: This study investigates the occurrence of miscarriages over the reproductive life span of women in a population-based study. STUDY DESIGN: A questionnaire was sent in 1994 to a random sample of 3000 Finnish women aged 18 to 44 years (73% response rate). Age-adjusted percentages of women having had miscarriages, and age- and other pregnancy event-adjusted odds ratios were calculated. RESULTS: Fifteen percent had at least 1 miscarriage. Miscarriages were more common in the moderately educated group and among women in health and social occupations, but there was no difference according to urbanism or health. By the age of 40 to 44 years, 90% of the women had been pregnant, and for only half had all pregnancies ended in a birth. The sequence of a miscarriage or miscarriages coming first and a birth or births coming later was more common than vice versa. CONCLUSION: In this population-based study miscarriages appeared incidental. Further studies on social class distribution might reveal new clues regarding etiology. PMID- 10454691 TI - Change in estimated cerebral perfusion pressure after treatment with nimodipine or magnesium sulfate in patients with preeclampsia. AB - OBJECTIVE: Data are accumulating to suggest that cerebral perfusion pressure may be either abnormally high or low in preeclampsia and eclampsia. Little is known of the cerebral perfusion pressure effects of magnesium sulfate or nimodipine. Our objective in this study was to compare the change in cerebral perfusion pressure in patients with severe preeclampsia randomly selected to receive nimodipine or magnesium sulfate. STUDY DESIGN: Patients with severe preeclampsia were randomly selected to receive magnesium sulfate (6 g bolus and 2 g/hr intravenous infusion) or nimodipine (60 mg taken orally every 4 hours). Transcranial Doppler ultrasonography was used to measure flow velocities in the right and left middle cerebral arteries, and the results were averaged. Measurements were obtained before treatment (baseline) and 30 minutes after the magnesium sulfate bolus was completely infused or 30 minutes after the nimodipine was ingested. Studies were performed before any other intervention. The person performing the ultrasonography was unaware of the patient's group assignment. Estimated cerebral perfusion pressure was calculated with the following formula: Estimated cerebral perfusion pressure = Velocity(mean) x [(Blood pressure(mean ) Blood pressure(diastolic ))/(Velocity(mean) - Velocity(diastolic ))]. The difference between estimated cerebral perfusion pressure at baseline and after treatment was compared between the 2 groups by means of the Mann-Whitney rank sum test. RESULTS: Nine patients were randomly selected to receive nimodipine and 12 to receive magnesium sulfate. Patient demographics and severity of condition were not significantly different between the 2 groups. The change in estimated cerebral perfusion pressure was significantly different between the groups. Estimated cerebral perfusion pressure increased after nimodipine use and decreased after magnesium sulfate use. CONCLUSION: Shortly after administration to patients with severe preeclampsia, nimodipine resulted in increased cerebral perfusion pressure in comparison with magnesium sulfate. PMID- 10454692 TI - Maternal neutrophil apoptosis in normal pregnancy, preeclampsia, and normotensive intrauterine growth restriction. AB - OBJECTIVE: In normal pregnancy there is both a neutrophilia and a mild neutrophil activation. In preeclampsia both direct and indirect evidence supports further marked neutrophil activation. In the pathogenesis of preeclampsia peripheral blood neutrophils may play a vital role in communicating between the preeclamptic placenta and the maternal vascular endothelium and contribute to the endothelial cell dysfunction that characterizes the maternal syndrome of preeclampsia. Preeclampsia shares many elements with the systemic inflammatory response syndrome. Neutrophils, key effectors of the systemic inflammatory response syndrome, are associated with hepatic necrosis and adult respiratory distress syndrome, both of which most commonly kill women with preeclampsia. We hypothesized that delayed neutrophil apoptosis could explain (1) the neutrophilia of normal pregnancy and (2) the differential maternal responses to the shared placental abnormality of preeclampsia and normotensive intrauterine growth restriction. STUDY DESIGN: Neutrophils were isolated (dextran 500, Ficoll [Amersham Pharmacia Biotech AB, Uppsala, Sweden], and erythrocyte lysis) from (1) case patients with preeclampsia at /=6). The analysis was performed separately for the 2 types of publications, and it was adjusted for the confounding effects of number of centers, funding status, and journal of publication. The beta coefficient (and its exponent) associated with the time term in the logistic model provided a measure of the trend in publication of multiauthored articles. RESULTS: In randomized studies the odds of publishing a multiauthored article, given the number of centers, funding status, and journal, were increased on average by 6% with every 5-year increment in time (odds ratio, 1.06; 95% confidence interval, 1.02-1. 10; P =.007). Similarly, in controlled observational studies, there was a 10% increase in the odds of publishing multiauthored articles (odds ratio, 1.10; 95% confidence interval, 1.01-1.20; P =.03). CONCLUSION: There is an inflationary trend in authorship that is not explained solely by the increased collaboration between centers and funding for research. PMID- 10454708 TI - Too little, too late. PMID- 10454709 TI - Collagen cross-linking in patients with urinary incontinence. PMID- 10454710 TI - Carpal tunnel syndrome and the myth of El Dorado. PMID- 10454711 TI - Isolated cranial nerve palsies due to brainstem lesions. AB - Isolated cranial nerve palsies are often attributed to lesions of the respective nerves along their extraaxial courses. There are a significant number of reports of individual patients with cranial nerve palsies, mostly of the 3rd and 6th nerves, as the sole manifestation of brainstem lesions proven by magnetic resonance imaging (MRI) or computer-assisted tomography (CT). An intraaxial basis may still be underestimated if based on MRI only, as electrophysiological abnormalities indicating brainstem lesions (masseter reflex, blink reflex, DC electrooculography) may be independent from MRI-documented morphological lesions. This article reviews the evidence that ischemic and demyelinating brainstem lesions are an important and underestimated cause of clinically isolated cranial nerve palsies. Especially in middle-aged and elderly people with 3rd and 6th nerve palsies, small pontine and mesencephalic infarctions seem to be more frequent than small-vessel ischemic infarctions of the extraaxial nerves. PMID- 10454712 TI - Sodium channels, excitability of primary sensory neurons, and the molecular basis of pain. AB - Following nerve injury, primary sensory neurons (dorsal root ganglion [DRG] neurons, trigeminal neurons) exhibit a variety of electrophysiological abnormalities, including increased baseline sensitivity and/or hyperexcitability, which can lead to abnormal burst activity that underlies pain, but the molecular basis for these changes has not been fully understood. Over the past several years, it has become clear that nearly a dozen distinct sodium channels are encoded by different genes and that at least six of these (including at least three distinct DRG- and trigeminal neuron-specific sodium channels) are expressed in primary sensory neurons. The deployment of different types of sodium channels in different types of DRG neurons endows them with different physiological properties. Dramatic changes in sodium channel expression, including downregulation of the SNS/PN3 and NaN sodium channel genes and upregulation of previously silent type III sodium channel gene, occur in DRG neurons following axonal transection. These changes in sodium channel gene expression are accompanied by a reduction in tetrodotoxin (TTX)-resistant sodium currents and by the emergence of a TTX-sensitive sodium current which recovers from inactivation (reprimes) four times more rapidly than the channels in normal DRG neurons. These changes in sodium channel expression poise DRG neurons to fire spontaneously or at inappropriately high frequencies. Changes in sodium channel gene expression also occur in experimental models of inflammatory pain. These observations indicate that abnormal sodium channel expression can contribute to the molecular pathophysiology of pain. They further suggest that selective blockade of particular subtypes of sodium channels may provide new, pharmacological approaches to treatment of disease involving hyperexcitability of primary sensory neurons. PMID- 10454713 TI - Motor unit action potential duration and muscle length. AB - Motor unit action potential (MUAP) components are investigated by means of single fiber computer simulations and clinical measurements. The single fiber simulations have essentially full bandwidth without noise, whereas the clinical measurements were made with a 3-10,000-Hz bandwidth utilizing approximately 1000 averages to reduce noise optimally. These parameters allow the recording of a MUAP's complete "physiologic" duration including its very slow onset and termination. The simulation results demonstrate a constant waveform onset regardless of the electrode's recording location along the fiber. A far-field potential is initiated when the action potential encounters the muscle fiber's termination. The simulated waveform's and clinically recorded MUAP's near-field component extends between the potential's onset and its corresponding far-field potential's onset. This near-field component's duration should vary with fiber length, and this prediction is clinically confirmed by measuring three different muscle lengths. The far-field potential reveals a constant duration, independent of fiber length, and appears to be associated with the muscle fiber's intracellular action potential duration. A more complete understanding of the components contributing to MUAP duration should provide a more fundamental basis for quantitative clinical MUAP duration measurements. PMID- 10454714 TI - Correlation between two-point discrimination and median nerve sensory response. AB - Static two-point discrimination (2PD) has been relied upon clinically to gauge the extent of median neuropathy in patients with carpal tunnel syndrome (CTS). Correlation with median nerve sensory conduction findings has not been well established. We determined the median sensory nerve action potential parameters from the first and third digits of 83 hands referred primarily for suspected CTS. These results were compared to 2PD carried out in a standardized fashion by a group of hand surgeons. A lack of correlation was found in most electrodiagnostic parameters, with the exception of peak and onset latencies to the thumb; on further analysis, 2PD to the thumb was found to be useful if abnormal, but contributed nothing if negative. We conclude that static 2PD results may correlate with latency, but do not overall adequately predict the findings on sensory nerve conduction examination of the median nerve. PMID- 10454715 TI - Facial nerve dysfunction in hereditary motor and sensory neuropathy type I and III. AB - Facial nerve function was studied in 19 patients with hereditary motor and sensory neuropathy type I (HMSN I) and 2 patients with hereditary motor and sensory neuropathy type III (HMSN III, Dejerine-Sottas), and compared to that in 24 patients with Guillain-Barre syndrome (GBS). The facial nerve was stimulated electrically at the stylomastoid fossa, and magnetically in its proximal intracanalicular segment. Additionally, the face-associated motor cortex was stimulated magnetically. The facial nerve motor neurography was abnormal in 17 of 19 HMSN I patients and in both HMSN III patients, revealing moderate to marked conduction slowing in both the extracranial and intracranial nerve segments, along with variable reductions of compound muscle action potential (CMAP) amplitudes. The facial nerve conduction slowing paralleled that of limb nerves, but was not associated with clinical dysfunction of facial muscles, because none of the HMSN I patients had facial palsy. Conduction slowing was most severe in the HMSN III patients, but only slight facial weakness was present. In GBS, conduction slowing was less marked, but facial weakness exceeded that in HMSN patients in all cases. We conclude that involvement of the facial nerve is common in HMSN I and HMSN III. It affects the intra- and extracranial part of the facial nerve and is mostly subclinical. PMID- 10454716 TI - Reorganization of flexion reflexes in the upper extremity of hemiparetic subjects. AB - We examined spatiotemporal abnormalities in the flexor reflex response in the impaired upper extremity of hemiparetic subjects. Electrical stimulation was used to elicit flexion reflexes in both upper extremities of 8 hemiparetic brain injured and 6 control subjects. Electromyograms (EMGs) were recorded from 12 arm muscles, and reflex forces and moments were recorded at the wrist with a load cell, and converted to shoulder and elbow torques. We found that the onset of reflex torque and EMG was delayed in the impaired arm and delays were greater at the shoulder than at the elbow. The normal reflex torque response consisted of elbow flexion, shoulder extension, and shoulder adduction. In contrast, in the impaired limb shoulder, flexion torque was observed in 7 subjects and shoulder abduction in 3. The delays in reflex onset and altered torque patterns in the impaired arm may be related to the abnormal movement synergies observed following stroke. © 1999 John Wiley & Sons, Inc. PMID- 10454717 TI - Ocular myasthenia gravis: predictive value of single-fiber electromyography. AB - Extraocular muscle weakness is the most common presenting sign of myasthenia gravis (MG). More than half of patients presenting with symptoms isolated to these muscles (OMG) develop generalized myasthenia gravis (GMG) over the course of their illness. No clinical, laboratory, or electrophysiological features are recognized that identify these high-risk patients. We have therefore assessed the ability of single-fiber electromyography (SFEMG) to predict the development of GMG in patients presenting with OMG. Thirty-nine consecutive patients presenting with OMG underwent SFEMG of the extensor digitorum communis muscle as well as a battery of other laboratory and imaging studies at the time of diagnosis. All patients were followed prospectively for a minimum of 24 months or until they developed GMG. Two patients were excluded, leaving 37 for assessment. Twenty remained with pure OMG for the entire follow-up period (mean, 55 months). Twenty six of the 37 had abnormal SFEMG studies at presentation. Eleven of these remained with OMG and 15 developed GMG. Fifty-eight percent of patients with an abnormal SFEMG developed GMG, whereas 82% of those with a normal study remained with OMG. Thus, a normal SFEMG was associated with MG remaining restricted to the extraocular muscles. (P = 0.036, Fisher's exact test), but an abnormal SFEMG was not predictive of subsequent development of GMG. PMID- 10454718 TI - Direct measurement of high-energy phosphate compounds in patients with neuromuscular disease. AB - Several neuromuscular disorders are associated with reductions in intramuscular adenosine triphosphate (ATP) and/or phosphocreatine (PCr). These alterations have been primarily characterized using (31)P-magnetic resonance spectroscopy ((31)P MRS). We prospectively measured total creatine, PCr, and ATP in muscle biopsies from 81 patients: normal controls (n = 33), mitochondrial cytopathy (n = 8), neuropathic (n = 3), dystrophy/congenital myopathies (n = 7), inflammatory myopathy (n = 12), and miscellaneous myopathies (n = 18) using direct biochemical analysis. Intramuscular concentrations of PCr and ATP were lower for the dystrophy/congenital myopathy, inflammatory myopathy, and mitochondrial disease patients with ragged red fiber (RRF) as compared with normal controls (P < 0.05). Total creatine was lower for the dystrophy/congenital myopathy group as compared with the normal control group (P < 0.05). These values compare favorably to results from other studies using (31)P-MRS and provide external validation for the values obtained using that method. Given the reductions in high-energy phosphate compounds in these patients, there is the potential for therapeutic intervention with creatine monohydrate supplementation. PMID- 10454719 TI - Lateral plantar neuropathy. AB - We report 8 cases of lateral plantar neuropathy (LPN). All had sensory impairment over the territory of the lateral plantar nerve. Near-nerve needle sensory nerve conduction study (NCS) of the plantar nerves showed abnormality confined to the lateral plantar nerve, confirming LPN. The most common cause for LPN was trauma and the most common site of injury was at the passage of the lateral plantar nerve through the abductor tunnel at the instep of the foot. PMID- 10454720 TI - Short-term aerobic training response in chronic myopathies. AB - We have previously demonstrated that patients with mitochondrial myopathies can benefit from short-term aerobic exercise training. In this study, we compared the responses to short-term aerobic training of patients with mitochondrial myopathies, patients with nonmetabolic myopathies, and sedentary normal subjects. Training consisted of 8 weeks of treadmill exercise at 70% to 85% of estimated maximum heart rate reserve. All groups showed significant improvements in estimated aerobic capacity as well as heart rate and blood lactate at submaximal exercise intensities. The increase in estimated aerobic capacity was greater in the mitochondrial myopathy patients than in the other two groups. Phosphorus magnetic resonance spectroscopy demonstrated increased oxidative capacity of muscle in patients with mitochondrial myopathies in response to this training but not in patients with other, nonmetabolic myopathies or sedentary control subjects. A self-assessed measurement of functional status (SF-36) suggested improved quality of life associated with the training. This study demonstrates that short-term aerobic training at low intensity can benefit patients with nonmetabolic myopathies but to a lesser extent than patients with mitochondrial myopathies. PMID- 10454721 TI - A comparison of two commercial quantitative electromyographic algorithms with manual analysis. AB - Quantitative EMG (QEMG) is a useful technique in the evaluation of neuromuscular disease. Manual waveform measurements have been replaced by automated computer based measurements, but there is no uniformity in computer algorithms used to make waveform measurements. We compared QEMG measurements made by algorithms in two commercially available EMG machines with manual measurements. Motor unit action potentials (MUAPs) were simultaneously fed into the two machines and analyzed using QEMG default settings and automatic waveform marking. The averaged MUAPs were also manually marked. The two algorithms and manual marking did not differ significantly for MUAP amplitude. There were significant differences between algorithms for duration and number of phases. Our study indicates that, although automated algorithms make QEMG more practical, visual inspection, and remarking of each MUAP if needed, is necessary before making clinical judgments from the data. PMID- 10454722 TI - Contractile properties of the quadriceps muscle in individuals with spinal cord injury. AB - Selected contractile properties and fatigability of the quadriceps muscle were studied in seven spinal cord-injured (SCI) and 13 able-bodied control (control) individuals. The SCI muscles demonstrated faster rates of contraction and relaxation than did control muscles and extremely large force oscillation amplitudes in the 10-Hz signal (65 +/- 22% in SCI versus 23 +/- 8% in controls). In addition, force loss and slowing of relaxation following repeated fatiguing contractions were greater in SCI compared with controls. The faster contractile properties and greater fatigability of the SCI muscles are in agreement with a characteristic predominance of fast glycolytic muscle fibers. Unexpectedly, the SCI muscles exhibited a force-frequency relationship shifted to the left, most likely as the result of relatively large twitch amplitudes. The results indicate that the contractile properties of large human locomotory muscles can be characterized using the approach described and that the transformation to faster properties consequent upon changes in contractile protein expression following SCI can be assessed. These measurements may be useful to optimize stimulation characteristics for functional electrical stimulation and to monitor training effects induced by electrical stimulation during rehabilitation of paralyzed muscles. PMID- 10454723 TI - Quantitative sudomotor axon reflex test (QSART): a new approach for testing distal sites. AB - The quantitative sudomotor axon reflex test (QSART) is a useful tool in the evaluation of C-fiber function. Because many neuropathies show a length-dependent loss of nerve function, a new capsule was designed for distal performance of the QSART, allowing measurements on fingers and toes. We investigated 20 healthy volunteers and 15 patients with insulin-dependent diabetes mellitus and normal sensory nerve conduction studies. A QSART was performed on the forearm and on the thumb. The groups showed no differences in the QSARTs on the forearm, but the sweat volume on the thumb was significantly lower in patients. Calculation of the forearm/finger ratio reduced the interindividual variation and showed the highest sensitivity for detecting C-fiber dysfunction in the patient group. We conclude that the distal measuring device and the forearm/finger ratio are more sensitive tools for the early detection of distal-onset neuropathies than are the commonly used proximal testing sites. PMID- 10454724 TI - Comparison of maximal voluntary isometric contraction and Drachman's hand-held dynamometry in evaluating patients with amyotrophic lateral sclerosis. AB - Maximal voluntary isometric contraction (MVIC) is a standard tool for assessment of muscle strength in treatment trials for amyotrophic lateral sclerosis (ALS). There is need for more practical bedside techniques especially for severely disabled patients. Hand-held dynamometry (HH-Dyn) is an inexpensive and easy-to handle device. MVIC was measured in five proximal muscle groups bilaterally and compared with HH-Dyn in 43 ALS patients. After a training period we found good intrarater correlation for HH-Dyn (r = 0.99), with a low coefficient of variation. Measurements tended to become more accurate after repeated testing due to practice effects in examiners and patients. Overall correlation between HH-Dyn and MVIC was good [r = 0.85 (P < 0.01)]. Strength-range-specific analysis showed a significant linear correlation up to 20 kg (44 lbs.) [r = 0.57 (P < 0.01)]. However, we found a tendency to underestimate muscle strength above 10 kg by HH Dyn as compared with MVIC, but this became meaningful only above a force of 20 kg. HH-Dyn provides a strength estimate with a precision close to MVIC in weak muscle groups (MRC grade 4). With standardization and appropriate training, HH Dyn is a useful bedside test, providing an alternative to MVIC for follow-up assessment in ALS. PMID- 10454726 TI - Sample size and the study of F waves. AB - Ulnar nerve F waves were studied in 23 healthy volunteers and 27 diabetic patients. Latencies and chronodispersion were analyzed in each group for different sample sizes. Significant differences were not detected with the different sample sizes for mean latencies, with samples above 16 stimuli or 10 waves for minimum and maximum latencies and above 20 stimuli or 16 waves for chronodispersion. These findings suggest that these limits may be adequate for group comparison. However, for the analysis of individual patients, the evidence suggests that larger samples are required for the determination of the minimum and maximum latencies and chronodispersion. PMID- 10454725 TI - Elongation of (CTG)n repeats in myotonic dystrophy protein kinase gene in tumors associated with myotonic dystrophy patients. AB - Length of (CTG)n triplet repeats in myotonic dystrophy protein kinase gene (DMPK) was estimated in tumors, normal tissues of the same organs, muscles, and leukocytes from three myotonic dystrophy (DM) patients and a non-DM patient. Using cDNA 25 as a probe, a Southern blot analysis of EcoRI- and BglI-digested DNA from these tissues demonstrated the longest expansion of the repeats in the tumors of DM patients. In all tissues from a non-DM patient, the repeat length was confirmed to be stable by PCR analysis. Our data suggest that expanded (CTG)n repeat in tumor tissues may have increased the instability. This study emphasizes the importance of a long-term prospective study on the incidence of tumors in DM to clarify the pathological interrelation between the two entities. PMID- 10454727 TI - Facial palsy in Heerfordt's syndrome: electrophysiological localization of the lesion. AB - Heerfordt's syndrome is characterized by fever, uveitis, swelling of the parotid gland, and facial nerve palsy and represents a variety of neurosarcoidosis. Since the first description of the syndrome, discussion about the lesion site has been controversial and has included the assumption of direct nerve compression by parotid gland swelling or a lesion within the facial canal in light of observations of accompanying taste disturbance. We report on a 26-year-old man with typical Heerfordt's syndrome who developed bilateral facial nerve palsy. Electrical and magnetic stimulation of the whole facial motor path provided strong evidence for a pathological process that: (i) began in the cerebellopontine angle; (ii) spread distally into the facial canal; and (iii) could be characterized by proximal demyelination. The patient recovered completely within 6 weeks under immunosuppressive therapy with steroids. PMID- 10454728 TI - Sympathetic skin responses in hereditary sensory and autonomic neuropathy and familial amyloid neuropathy are different. AB - We compared the clinical and electrophysiological findings in seven patients with familial amyloid polyneuropathy (FAP) and six with hereditary sensory and autonomic neuropathy type I (HSAN). Both groups had progressive loss of pain and temperature appreciation, beginning distally in the feet. In HSAN, the median sensory nerve action potentials (SNAP) were lost early, but the sympathetic skin responses (SSR) were always preserved. In FAP, the SSR were lost at an early stage. The SSR may thus help to distinguish between these two causes of small fiber neuropathy early in the course of the disorder. PMID- 10454729 TI - Cervical root stimulation in a case of classic neurogenic thoracic outlet syndrome. AB - We performed C8 nerve root stimulation in addition to other electromyographic (EMG) studies in a surgically proven case of classic thoracic outlet syndrome (TOS). The patient was a 19-year-old woman with a 2-year history of right hand cramps and progressive weakness and atrophy of hand muscles, especially the thenar eminence. Routine EMG studies showed evidence for an axon-loss lower trunk brachial plexopathy. Stimulation studies of the C8 nerve roots demonstrated proximal conduction block on the affected side only. The diagnosis was further supported by cervical spine radiographs, which demonstrated a cervical rib, and surgical exploration of the brachial plexus, which demonstrated upward compression and stretching of the lower trunk by a fascial band extending from the anomalous cervical rib to the first thoracic rib. The patient noted a modest improvement in hand function postoperatively. Root stimulation studies can help in the diagnosis of classic TOS by providing more precise localization and information regarding the degree, if any, of proximal motor conduction block. PMID- 10454730 TI - Neuromuscular transmission defect caused by carbamazepine. AB - The clinical effect of carbamazepine (CBZ) on neuromuscular transmission is described in two children who presented in coma with diffuse hypotonia and areflexia following CBZ overdose. Repetitive nerve stimulation (RNS) showed a decremental response only at high-frequency stimulation. With supportive care, the patients made an uneventful recovery. Follow-up RNS was normal. This is the first report of a clinically evident neuromuscular transmission defect produced by CBZ. We postulate that CBZ's known effect on decreasing sodium channel depolarization produced a defect in neuromuscular transmission. The report emphasizes the contribution of RNS in the evaluation of coma of uncertain etiology, particularly in cases of possible intoxication, and the potential for CBZ to compromise neuromuscular transmission in normal individuals or in patients with a decreased neuromuscular transmission safety factor. PMID- 10454731 TI - Myasthenia gravis in the tenth decade. PMID- 10454732 TI - Immunostaining of dystrophin using catalyzed signal amplification. PMID- 10454733 TI - Supplementation with creatine monohydrate in children with mitochondrial encephalomyopathies. PMID- 10454734 TI - Abstracts of presentations: 46th annual scientific meeting american association of electrodiagnostic medicine PMID- 10454735 TI - Medicolegal issues in fertility regulation. PMID- 10454736 TI - Control of CD4 gene expression: connecting signals to outcomes in T cell development. AB - The control of CD4 gene expression is essential for proper T lymphocyte development. Signals transmitted from the T-cell antigen receptor (TCR) during the thymic selection processes are believed to be linked to the regulation of CD4 gene expression during specific stages of T cell development. Thus, a study of the factors that control CD4 gene expression may lead to further insight into the molecular mechanisms that drive thymic selection. In this review, we discuss the work conducted to date to identify and characterize the cis-acting transcriptional control elements in the CD4 locus and the DNA-binding factors that mediate their function. From these studies, it is becoming clear that the molecular mechanisms controlling CD4 gene expression are very complex and differ at each stage of development. Thus, the control of CD4 expression is subject to many different influences as the thymocyte develops. PMID- 10454737 TI - Functions of the extracellular matrix and matrix degrading proteases during tumor progression. AB - Cell interactions with extracellular matrices are important to pathological changes that occur during cell transformation and tumorigenesis. Several extracellular matrix proteins including fibronectin, thrombospondin-1, laminin, SPARC, and osteopontin have been suggested to modulate tumor phenotype by affecting cell migration, survival, or angiogenesis. Likewise, proteases including the matrix metalloproteinases (MMPs) are understood to not only facilitate migration of cells by degradation of matrices, but also to affect tumor formation and growth. We have recently demonstrated an in vivo role for the RGD-containing protein, osteopontin, during tumor progression, and found evidence for distinct functions in the host versus the tumor cells. Because of the compartmentalization and temporal regulation of MMP expression, it is likely that MMPs may also function dually in host stroma and the tumor cell. In addition, an important function of proteases appears to be not only degradation, but also cleavage of matrix proteins to generate functionally distinct fragments based on receptor binding, biological activity, or regulation of growth factors. PMID- 10454738 TI - Nuclear exclusion of transcription factors associated with apoptosis in developing nervous tissue. AB - Programmed cell death in the form of apoptosis involves a network of metabolic events and may be triggered by a variety of stimuli in distinct cells. The nervous system contains several neuron and glial cell types, and developmental events are strongly dependent on selective cell interactions. Retinal explants have been used as a model to investigate apoptosis in nervous tissue. This preparation maintains the structural complexity and cell interactions similar to the retina in situ, and contains cells in all stages of development. We review the finding of nuclear exclusion of several transcription factors during apoptosis in retinal cells. The data reviewed in this paper suggest a link between apoptosis and a failure in the nucleo-cytoplasmic partition of transcription factors. It is argued that the nuclear exclusion of transcription factors may be an integral component of apoptosis both in the nervous system and in other types of cells and tissues. PMID- 10454739 TI - Perturbation of EGF-induced MAP kinase activation by TGF-beta 1. AB - TGF-beta 1 regulates both cellular growth and phenotypic plasticity important for maintaining a growth advantage and increased invasiveness in progressively malignant cells. Recent studies indicate that TGF-beta-1 stimulates the conversion of epitheliod to fibroblastoid phenotype which presumably leads to the inactivation of growth-inhibitory effects by TGF-beta 1 (Portella et al. (1998) Cell Growth and Differentiation, 9: 393-404). Therefore, the investigation of TGF beta 1 signaling that leads to altered growth and migration may provide novel targets for the prevention of increased cell growth and invasion. Although much attention has been paid to TGF-beta 1 responses in epithelial cells, the above studies suggest that examination of signal transduction pathways in fibroblasts are important as well. Data from our laboratory are consistent with the concept that TGF-beta 1 can act as a regulatory switch in density-dependent C3H 10T1/2 fibroblasts capable of either promoting or delaying G1 traverse. The regulation of this switch is proposed to occur prior to pRb phosphorylation, namely prior to activation of cyclin-dependent kinases. The current study is concerned with the evaluation of a key cyclin (cyclin D1) which activates cdk4 and p27KIP1 which in turn inhibit cdk2 in the proliferative responses of epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) and their modulation by TGF-beta 1. Although the molecular events that lead to elevation of cyclin D1 are not completely understood, it appears likely that activation of p42/p44MAPK kinases is involved in its transcriptional regulation. TGF-beta 1 delayed EGF- or PDGF induced cyclin D1 expression and blocked the induction of active p42/p44MAPK. The mechanism by which TGF-beta 1 induces a block in p42/p44MAPK activation is being examined and the possibility that TGF-beta 1 regulates phosphatase activity is being tested. PMID- 10454740 TI - Collateral methotrexate resistance in cisplatin-selected murine leukemia cells. AB - Resistance to anticancer drugs is a major cause of failure of many therapeutic protocols. A variety of mechanisms have been proposed to explain this phenomenon. The exact mechanism depends upon the drug of interest as well as the tumor type treated. While studying a cell line selected for its resistance to cisplatin we noted that the cells expressed a > 25,000-fold collateral resistance to methotrexate. Given the magnitude of this resistance we elected to investigate this intriguing collateral resistance. From a series of investigations we have identified an alteration in a membrane protein of the resistant cell as compared to the sensitive cells that could be the primary mechanism of resistance. Our studies reviewed here indicate decreased tyrosine phosphorylation of a protein (molecular mass = 66) in the resistant cells, which results in little or no transfer of methotrexate from the medium into the cell. Since this is a relatively novel function for tyrosine phosphorylation, this information may provide insight into possible pharmacological approaches to modify therapeutic regimens by analyzing the status of this protein in tumor samples for a better survival of the cancer patients. PMID- 10454741 TI - Searching for the role of protein phosphatases in eukaryotic microorganisms. AB - Preference for specific protein substrates together with differential sensitivity to activators and inhibitors has allowed classification of serine/threonine protein phosphatases (PPs) into four major types designated types 1, 2A, 2B and 2C (PP1, PP2A, PP2B and PP2C, respectively). Comparison of sequences within their catalytic domains has indicated that PP1, PP2A and PP2B are members of the same gene family named PPP. On the other hand, the type 2C enzyme does not share sequence homology with the PPP members and thus represents another gene family, known as PPM. In this report we briefly summarize some of our studies about the role of serine/threonine phosphatases in growth and differentiation of three different eukaryotic models: Blastocladiella emersonii, Neurospora crassa and Dictyostelium discoideum. Our observations suggest that PP2C is the major phosphatase responsible for dephosphorylation of amidotransferase, an enzyme that controls cell wall synthesis during Blastocladiella emersonii zoospore germination. We also report the existence of a novel acid- and thermo-stable protein purified from Neurospora crassa mycelia, which specifically inhibits the PP1 activity of this fungus and mammals. Finally, we comment on our recent results demonstrating that Dictyostelium discoideum expresses a gene that codes for PP1, although this activity has never been demonstrated biochemically in this organism. PMID- 10454742 TI - Control of the adrenocortical cell cycle: interaction between FGF2 and ACTH. AB - FGF2 elicits a strong mitogenic response in the mouse Y-1 adrenocortical tumor cell line, that includes a rapid and transient activation of the ERK-MAPK cascade and induction of the c-Fos protein. ACTH, itself a very weak mitogen, blocks the mitogenic response effect of FGF2 in the early and middle G1 phase, keeping both ERK-MAPK activation and c-Fos induction at maximal levels. Probing the mitogenic response of Y-1 cells to FGF2 with ACTH is likely to uncover reactions underlying the effects of this hormone on adrenocortical cell growth. PMID- 10454743 TI - Application of the differential display RT-PCR strategy for the identification of inflammation-related mouse genes. AB - The inflammatory response elicited by various stimuli such as microbial products or cytokines is determined by differences in the pattern of cellular gene expression. We have used the differential display RT-PCR (DDRT-PCR) strategy to identify mRNAs that are differentially expressed in various murine cell types stimulated with pro-inflammatory cytokines, microbial products or anti inflammatory drugs. Mouse embryonic fibroblasts (MEFs) were treated with IFNs, TNF, or sodium salicylate. Also, peritoneal macrophages from C3H/Hej mice were stimulated with T. cruzi-derived GPI-mucin and/or IFN-gamma. After DDRT-PCR, various cDNA fragments that were differentially represented on the sequencing gel were recovered, cloned and sequenced. Here, we describe a summary of several experiments and show that, when 16 of a total of 28 recovered fragments were tested for differential expression, 5 (31%) were found to represent mRNAs whose steady-state levels are indeed modulated by the original stimuli. Some of the identified cDNAs encode for known proteins that were not previously associated with the inflammatory process triggered by the original stimuli. Other cDNA fragments (8 of 21 sequences, or 38%) showed no significant homology with known sequences and represent new mouse genes whose characterization might contribute to our understanding of inflammation. In conclusion, DDRT-PCR has proven to be a potent technology that will allow us to identify genes that are differentially expressed when cells are subjected to changes in culture conditions or isolated from different organs. PMID- 10454744 TI - A receptor for infectious and cellular prion protein. AB - Prions are an unconventional form of infectious agents composed only of protein and involved in transmissible spongiform encephalopathies in humans and animals. The infectious particle is composed by PrPsc which is an isoform of a normal cellular glycosyl-phosphatidylinositol (GPI) anchored protein, PrPc, of unknown function. The two proteins differ only in conformation, PrPc is composed of 40% alpha helix while PrPsc has 60% beta-sheet and 20% alpha helix structure. The infection mechanism is trigged by interaction of PrPsc with cellular prion protein causing conversion of the latter's conformation. Therefore, the infection spreads because new PrPsc molecules are generated exponentially from the normal PrPc. The accumulation of insoluble PrPsc is probably one of the events that lead to neuronal death. Conflicting data in the literature showed that PrPc internalization is mediated either by clathrin-coated pits or by caveolae-like membranous domains. However, both pathways seem to require a third protein (a receptor or a prion-binding protein) either to make the connection between the GPI-anchored molecule to clathrin or to convert PrPc into PrPsc. We have recently characterized a 66-kDa membrane receptor which binds PrPc in vitro and in vivo and mediates the neurotoxicity of a human prion peptide. Therefore, the receptor should have a role in the pathogenesis of prion-related diseases and in the normal cellular process. Further work is necessary to clarify the events triggered by the association of PrPc/PrPsc with the receptor. PMID- 10454745 TI - Mechanisms of cell transformation induced by polyomavirus. AB - Polyomavirus is a DNA tumor virus that induces a variety of tumors in mice. Its genome encodes three proteins, namely large T (LT), middle T (MT), and small T (ST) antigens, that have been implicated in cell transformation and tumorigenesis. LT is associated with cell immortalization, whereas MT plays an essential role in cell transformation by binding to and activating several cytoplasmic proteins that participate in growth factor-induced mitogenic signal transduction to the nucleus. The use of different MT mutants has led to the identification of MT-binding proteins as well as analysis of their importance during cell transformation. Studying the molecular mechanisms of cell transformation by MT has contributed to a better understanding of cell cycle regulation and growth control. PMID- 10454746 TI - Functional analysis of newly discovered growth control genes: experimental approaches. AB - A large number of DNA sequences corresponding to human and animal transcripts have been filed in data banks, as cDNAs or ESTs (expression sequence tags). However, the actual function of their corresponding gene products is still largely unknown. Several of these genes may play a role in regulation of important biological processes such as cell division, differentiation, malignant transformation and oncogenesis. Elucidation of gene function is based on 2 main approaches, namely, overexpression and expression interference, which respectively mimick or suppress a given phenotype. The currently available tools and experimental approaches to gene functional analysis and the most recent advances in mass cDNA screening by functional analysis are discussed. PMID- 10454747 TI - Hunting for differentially expressed genes. AB - Differentially expressed genes are usually identified by comparing steady-state mRNA concentrations. Several methods have been used for this purpose, including differential hybridization, cDNA subtraction, differential display and, more recently, DNA chips. Subtractive hybridization has significantly improved after the polymerase chain reaction was incorporated into the original method and many new protocols have been established. Recently, the availability of the wellknown coding sequences for some organisms has greatly facilitated gene expression analysis using high-density microarrays. Here, we describe some of these modifications and discuss the benefits and drawbacks of the various methods corresponding to the main advances in this field. PMID- 10454748 TI - Collagenase specificity in chondrosarcoma metastasis. AB - The treatment of some mesenchymal malignancies has made significant gains over the past few decades with the development of effective systemic therapies. In contrast, the treatment of chondrosarcoma has been limited to surgical resection, with the most significant prognostic indicators being surgical margins and histologic grade. We have reported that MMP-1/TIMP-1 gene expression serves to prognosticate for tumor recurrence in this group of patients. This led to the hypothesis that collagenase activity facilitates cell egression from the cartilaginous matrix. In the current study we examine the specificity of collagenase gene expression in archival human chondrosarcoma samples using semi quantitative PCR. Messenger RNA was affinity extracted and subject to reverse transcription. The subsequent cDNA was amplified using novel primers and quantitated by densitometry. Ratios of gene expression were constructed and compared to disease-free survival. The data demonstrate that the significance of the MMP-1/TIMP-1 ratio as a predictor of recurrence is confirmed with a larger number of patients. Neutrophil collagenase or MMP-8 was observed in only 5 of 29 samples. Collagenase-3 or MMP-13 was observed in all samples but the level did not correlate with disease-free survival. Since the collagenases have similar activity for fibrillar collagens and cleave the peptide in the same location, post-transcriptional regulatory mechanisms may account for the observed specificity. The determination of the MMP-1/TIMP-1 gene expression ratio not only serves to identify those patients at risk for recurrence but may also serve as a novel therapeutic avenue as an adjunct to surgical resection. PMID- 10454749 TI - Oncogene-mediated downregulation of RECK, a novel transformation suppressor gene. AB - The RECK gene was initially isolated as a transformation suppressor gene encoding a novel membrane-anchored glycoprotein and later found to suppress tumor invasion and metastasis by regulating matrix metalloproteinase-9. Its expression is ubiquitous in normal tissues, but undetectable in many tumor cell lines and in fibroblastic lines transformed by various oncogenes. The RECK gene promoter has been cloned and characterized. One of the elements responsible for the oncogene mediated downregulation of mouse RECK gene is the Sp1 site, where the Sp1 and Sp3 factors bind. Sp1 transcription factor family is involved in the basal level of promoter activity of many genes, as well as in dynamic regulation of gene expression; in a majority of cases as a positive regulator, or, as exemplified by the oncogene-mediated suppression of RECK gene expression, as a negative transcription regulator. The molecular mechanisms of the down-regulation of mouse RECK gene and other tumor suppressor genes are just beginning to be uncovered. Understanding the regulation of these genes may help to develop strategies to restore their expression in tumor cells and, hence, suppress the cells' malignant behavior. PMID- 10454750 TI - Genetic alterations in head and neck squamous cell carcinomas. AB - The genetic alterations observed in head and neck cancer are mainly due to oncogene activation (gain of function mutations) and tumor suppressor gene inactivation (loss of function mutations), leading to deregulation of cell proliferation and death. These genetic alterations include gene amplification and overexpression of oncogenes such as myc, erbB-2, EGFR and cyclinD1 and mutations, deletions and hypermethylation leading to p16 and TP53 tumor suppressor gene inactivation. In addition, loss of heterozygosity in several chromosomal regions is frequently observed, suggesting that other tumor suppressor genes not yet identified could be involved in the tumorigenic process of head and neck cancers. The exact temporal sequence of the genetic alterations during head and neck squamous cell carcinoma (HNSCC) development and progression has not yet been defined and their diagnostic or prognostic significance is controversial. Advances in the understanding of the molecular basis of head and neck cancer should help in the identification of new markers that could be used for the diagnosis, prognosis and treatment of the disease. PMID- 10454751 TI - Efficient retrovirus-mediated transfer of cell-cycle control genes to transformed cells. AB - The use of gene therapy continues to be a promising, yet elusive, alternative for the treatment of cancer. The origins of cancer must be well understood so that the therapeutic gene can be chosen with the highest chance of successful tumor regression. The gene delivery system must be tailored for optimum transfer of the therapeutic gene to the target tissue. In order to accomplish this, we study models of G1 cell-cycle control in both normal and transformed cells in order to understand the reasons for uncontrolled cellular proliferation. We then use this information to choose the gene to be delivered to the cells. We have chosen to study p16, p21, p53 and pRb gene transfer using the pCL-retrovirus. Described here are some general concepts and specific results of our work that indicate continued hope for the development of genetically based cancer treatments. PMID- 10454752 TI - Genetic susceptibility to HPV infection and cervical cancer. AB - Squamous cell carcinoma of the cervix (SCCC) is one of the leading causes of death in developing countries. Infection with high-risk human papillomavirus (HPV) is the major risk factor to develop malignant lesions in the cervix. Polymorphisms of the MHC and p53 genes seem to influence the outcome of HPV infection and progression to SCCC, although controversial data have been reported. MHC are highly polymorphic genes that encode molecules involved in antigen presentation, playing a key role in immune regulation, while p53 is a tumor suppressor gene that regulates cell proliferation. The HPV E6 protein from high-risk types binds p53 and mediates its degradation by the ubiquitin pathway. The role of these polymorphisms in genetic susceptibility to HPV infection and to SCCC remains under investigation. PMID- 10454753 TI - P-glycoprotein-mediated resistance to chemotherapy in cancer cells: using recombinant cytosolic domains to establish structure-function relationships. AB - Resistance to chemotherapy in cancer cells is mainly mediated by overexpression of P-glycoprotein (Pgp), a plasma membrane ATP-binding cassette (ABC) transporter which extrudes cytotoxic drugs at the expense of ATP hydrolysis. Pgp consists of two homologous halves each containing a transmembrane domain and a cytosolic nucleotide-binding domain (NBD) which contains two consensus Walker motifs, A and B, involved in ATP binding and hydrolysis. The protein also contains an S signature characteristic of ABC transporters. The molecular mechanism of Pgp mediated drug transport is not known. Since the transporter has an extraordinarily broad substrate specificity, its cellular function has been described as a "hydrophobic vacuum cleaner". The limited knowledge about the mechanism of Pgp, partly due to the lack of a high-resolution structure, is well reflected in the failure to efficiently inhibit its activity in cancer cells and thus to reverse multidrug resistance (MDR). In contrast to the difficulties encountered when studying the full-length Pgp, the recombinant NBDs can be obtained in large amounts as soluble proteins. The biochemical and biophysical characterization of recombinant NBDs is shown here to provide a suitable alternative route to establish structure-function relationships. NBDs were shown to bind ATP and analogues as well as potent modulators of MDR, such as hydrophobic steroids, at a region close to the ATP site. Interestingly, flavonoids also bind to NBDs with high affinity. Their binding site partly overlaps both the ATP-binding site and the steroid-interacting region. Therefore flavonoids constitute a new promising class of bifunctional modulators of Pgp. PMID- 10454754 TI - Arylsulfatase A pseudodeficiency in healthy Brazilian individuals. AB - Molecular alterations associated with arylsulfatase A pseudodeficiency (ASA-PD) were characterized by PCR and restriction endonuclease analysis in a sample of healthy individuals from Brazil. ASA activity was also assayed in all subjects. Two individuals homozygous for the N350S and 1524+95A<--G mutations were detected, corresponding to a frequency of 1.17% (4 of 324 alleles). The individual frequency of the N350S mutation was 20.7% (71 of 342 alleles) and 7.9% (27 of 342 alleles) for the 1524+95A<--G mutation. The frequency of the ASA-PD allele in our population was estimated to be 7.9%. This is the first report of ASA-PD allele frequency in a South American population. In addition, the methods used are effective and suitable for application in countries with limited resources. All patients with low ASA activity should be screened for ASA-PD as part of the diagnostic protocol for metachromatic leukodystrophy. PMID- 10454755 TI - Hydrolysis of xylans by enzyme systems from solid cultures of Trichoderma harzianum strains. AB - Xylanase activity was isolated from crude extracts of Trichoderma harzianum strains C and 4 grown at 28 degree C in a solid medium containing wheat bran as the carbon source. Enzyme activity was demonstrable in the permeate after ultrafiltration of the crude extracts using an Amicon system. The hydrolysis patterns of different xylans and paper pulps by xylanase activity ranged from xylose, xylobiose and xylotriose to higher xylooligosaccharides. A purified ss xylosidase from the Trichoderma harzianum strain released xylose, xylobiose and xylotriose from seaweed, deacetylated, oat spelt and birchwood xylans. The purified enzyme was not active against acetylated xylan and catalyzed the hydrolysis of xylooligosaccharides, including xylotriose, xylotetraose and xylopentaose. However, the enzyme was not able to degrade xylohexaose. Xylanase pretreatment was effective for hardwood kraft pulp bleaching. Hardwood kraft pulp bleached in the XEOP sequence had its kappa number reduced from 13.2 to 8.9 and a viscosity of 20. 45 cp. The efficiency of delignification was 33%. PMID- 10454756 TI - Detection of cytomegalovirus infections by PCR in renal transplant patients. AB - Cytomegalovirus (CMV) is the single most important infectious agent affecting recipients of organ transplants. To evaluate the incidence and the clinical importance of CMV infection in renal transplants in Brazil, 37 patients submitted to renal allograft transplants were tested periodically for the presence of cytomegalovirus DNA in urine using the polymerase chain reaction (PCR), and for the presence of IgM and IgG antibodies against CMV by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence (IIF). The PCR-amplified products were detected by gel electrophoresis and confirmed by dot-blot hybridization with oligonucleotide probes. Thirty-two of the 37 patients (86.4%) were positive by at least one of the three methods. In six patients, PCR was the only test which detected the probable CMV infection. Ten patients had a positive result by PCR before transplantation. In general, the diagnosis was achieved earlier by PCR than by serologic tests. Active infection occurred more frequently during the first four months after transplantation. Sixteen of the 32 patients (50%) with active CMV infection presented clinical symptoms consistent with CMV infection. Five patients without evidence of active CMV infection by the three tests had only minor clinical manifestations during follow-up. Our results indicate that PCR is a highly sensitive procedure for the early detection of CMV infection and that CMV infection in renal transplant patients is a frequent problem in Brazil. PMID- 10454757 TI - Enhanced mucosal re-epithelialization induced by short chain fatty acids in experimental colitis. AB - The short chain fatty acids (SCFA) are the best nutrients for the colonocytes. Glucose is poorly used as a fuel but may be transformed into SCFA by colonic bacteria. The aim of this study was to investigate the effect of SCFA or glucose on experimental colitis. Colitis was induced in 30 Wistar rats by colonic instillation of 4% acetic acid. Five days later they were randomized to receive twice a day colonic lavage containing saline (controls, N = 10), 10% hypertonic glucose (N = 10) or SCFA (N = 10) until day 8 when they were killed. At autopsy, the colon was removed and weighed and the mucosa was evaluated macro- and microscopically and stripped out for DNA assay. Data are reported as mean +/- SD or median [range] as appropriate. All animals lost weight but there was no difference between groups. Colon weight was significantly lower in the SCFA group (3.8 +/- 0.5 g) than in the control (5.3 +/- 2.1 g) and glucose (5.2 +/- 1.3 g) groups (P<0.05). Macroscopically, the severity of inflammation was less in SCFA (grade 2 [1-5]) than in control (grade 9 [4-10]) and glucose-treated (grade 9 [2 10]) animals (P<0.01). Microscopically, ulceration of the mucosa was more severe in the glucose and control groups than in the SCFA group. The DNA content of the mucosa of SCFA-treated animals (8.2 [5.0-20.2] mg/g of tissue) was higher than in glucose-treated (5.1 [4.2-8.5] mg/g of tissue; P<0.01) and control (6.2 [4.5-8.9] mg/g of tissue; P<0.05) animals. We conclude that SCFA may enhance mucosal re epithelialization in experimental colitis, whereas hypertonic glucose is of no benefit. PMID- 10454758 TI - Evaluation of an anti-carcinoembryonic monoclonal antibody suitable for immunoscintigraphy. AB - An anti-carcinoembryonic antigen (CEA) monoclonal antibody (mAb 6D1. 1) was evaluated in vitro and in vivo to determine its suitability as a tracer for immunoscintigraphy of colorectal carcinomas. Determination of mAb affinity for CEA showed a constant of association of 0.63 +/- 0.11 x 10(9) M-1. Binding of technetium-99m (99mTc)-6D1.1, labeled by a direct method, to human cultured lineages was highly specific. Binding to only CEA-positive LS-174T cells resulted in a saturable curve inhibited by pre-incubation with unlabeled mAb. No binding at all was observed for the human lineages MeWo (melanoma) or ZR75-30 (breast carcinoma), neither of them expressing CEA cells. Intravenous injection of 99mTc 6D1.1 into nude mice xenografted with human LS-174T tumors resulted in planar images of excellent quality. Localization of an irrelevant mAb labeled with either 99mTc or iodine-125 was never observed in tumor masses. Biodistribution studies on excised tumoral tissue showed retention of 28.48% of the injected dose per gram of LS-174T tumor. The tumor-to-blood ratio was 3.46. The same analysis performed on the other three human xenografted tumors studied demonstrated that only the CEA-producing HT-29 (colorectal adenocarcinoma) retained 99mTc-6D1.1 while the other two (ZR75-30 and MeWo) did not. These data demonstrate that this mAb is an adequate tool for targeting CEA-expressing tumors in experimental models. PMID- 10454759 TI - Ethological analysis of mother-pup interactions and other behavioral reactions in rats: effects of malnutrition and tactile stimulation of the pups. AB - Mother-pup interaction, as well as other behavioral reactions were studied during the lactation period in 24 litters of Wistar rats and their dams fed either a 16% (control - C; 12 litters) or a 6% (malnourished - M; 12 litters) protein diet. The diets were isocaloric. Throughout lactation there was a 36.4% weight loss of M dams and a 63% body weight deficit in the M pups when compared to control pups. During this period, half of the litters were exposed daily to additional tactile stimulation (CS or MS), while the other half were submitted to normal rearing conditions (CN or MN). The tactile stimulation of pups (handling) consisted of holding the animal in one hand and gently touching the dorsal part of the animal's body with the fingers for 3 min. A special camera and a time-lapse video were used to record litter behavior in their home cages. Starting at 6 p.m. and ending at 6 a.m., on days 3, 6, 12, 15, 18 and 21 of lactation, photos were taken at 4-s intervals. An increase in the frequency (154.88 +/- 16.19) and duration (455.86 +/- 18.05 min) of suckling was observed throughout the lactation period in all groups compared to birth day (frequency 24.88 +/- 2.37 and duration 376.76 +/- 21.01 min), but the frequency was higher in the C (84.96 +/- 8.52) than in the M group (43.13 +/- 4.37); however, the M group (470.2 +/- 11.87 min) spent more time suckling as compared with the C group (393.67 +/- 13.09 min). The M dams showed a decreased frequency of resting position throughout the lactation period (6.5 +/- 2.48) compared to birth day (25.42 +/- 7.74). Pups from the C group were more frequently observed separated (73.02 +/- 4.38) and interacting (258.99 +/- 20.61) more with their mothers than the M pups (separated 66.94 +/- 5.5 and interacting 165.72 +/- 12.05). Tactile stimulation did not interact with diet condition, showing that the kind of stimulation used in the present study did not lead to recovery from the changes induced by protein malnutrition. The changes in mother-pup interaction produced by protein malnutrition of both may represent retardation in neuromotor development and a higher dependence of the pups on their mothers. These changes may represent an important means of energy saving and heat maintenance in malnourished pups. PMID- 10454760 TI - Effects of microcystin-LR in isolated perfused rat kidney. AB - Microcystin is a hepatotoxic peptide which inhibits protein phosphatase types 1 and 2A. The objective of the present study was to evaluate the physiopathologic effects of microcystin-LR in isolated perfused rat kidney. Adult Wistar rats (N = 5) of both sexes (240-280 g) were utilized. Microcystin-LR (1 microg/ml) was perfused over a period of 120 min, during which samples of urine and perfusate were collected at 10-min intervals to determine the levels of inulin, sodium, potassium and osmolality. We observed a significant increase in urinary flow with a peak effect at 90 min (control (C) = 0.20 +/- 0.01 and treated (T) = 0.32 +/- 0.01 ml g-1 min-1, P<0.05). At 90 min there was a significant increase in perfusate pressure (C = 129.7 +/- 4.81 and T = 175.0 +/- 1.15 mmHg) and glomerular filtration rate (C = 0.66 +/- 0.07 and T = 1.10 +/- 0. 04 ml g-1 min 1) and there was a significant reduction in fractional sodium tubular transport at 120 min (C = 78.6 +/- 0.98 and T = 73.9 +/- 0.95%). Histopathologic analysis of the perfused kidneys showed protein material in the urinary space, suggestive of renal toxicity. These data demonstrate renal vascular, glomerular and urinary effects of microcystin-LR, indicating that microcystin acts directly on the kidney by probable inhibition of protein phosphatases. PMID- 10454761 TI - Effects of mercury on the arterial blood pressure of anesthetized rats. AB - The available data suggests that hypotension caused by Hg2+ administration may be produced by a reduction of cardiac contractility or by cholinergic mechanisms. The hemodynamic effects of an intravenous injection of HgCl2 (5 mg/kg) were studied in anesthetized rats (N = 12) by monitoring left and right ventricular (LV and RV) systolic and diastolic pressures for 120 min. After HgCl2 administration the LV systolic pressure decreased only after 40 min (99 +/- 3.3 to 85 +/- 8.8 mmHg at 80 min). However, RV systolic pressure increased, initially slowly but faster after 30 min (25 +/- 1.8 to 42 +/- 1.6 mmHg at 80 min). Both right and left diastolic pressures increased after HgCl2 treatment, suggesting the development of diastolic ventricular dysfunction. Since HgCl2 could be increasing pulmonary vascular resistance, isolated lungs (N = 10) were perfused for 80 min with Krebs solution (continuous flow of 10 ml/min) containing or not 5 microM HgCl2. A continuous increase in pulmonary vascular resistance was observed, suggesting the direct effect of Hg2+ on the pulmonary vessels (12 +/- 0.4 to 29 +/- 3.2 mmHg at 30 min). To examine the interactions of Hg2+ and changes in cholinergic activity we analyzed the effects of acetylcholine (Ach) on mean arterial blood pressure (ABP) in anesthetized rats (N = 9) before and after Hg2+ treatment (5 mg/kg). Using the same amount and route used to study the hemodynamic effects we also examined the effects of Hg2+ administration on heart and plasma cholinesterase activity (N = 10). The in vivo hypotensive response to Ach (0.035 to 10.5 microg) was reduced after Hg2+ treatment. Cholinesterase activity (microM h-1 mg protein-1) increased in heart and plasma (32 and 65%, respectively) after Hg2+ treatment. In conclusion, the reduction in ABP produced by Hg2+ is not dependent on a putative increase in cholinergic activity. HgCl2 mainly affects cardiac function. The increased pulmonary vascular resistance and cardiac failure due to diastolic dysfunction of both ventricles are factors that might contribute to the reduction of cardiac output and the fall in arterial pressure. PMID- 10454762 TI - Melatonin modulation of presynaptic nicotinic acetylcholine receptors located on short noradrenergic neurons of the rat vas deferens: a pharmacological characterization. AB - Melatonin, the pineal hormone produced during the dark phase of the light-dark cycle, modulates neuronal acetylcholine receptors located presynaptically on nerve terminals of the rat vas deferens. Recently we showed the presence of high affinity nicotine-binding sites during the light phase, and low and high affinity binding sites during the dark phase. The appearance of the low affinity binding sites was due to the nocturnal melatonin surge and could be mimicked by exposure to melatonin in vitro. The aim of the present research was to identify the receptor subtypes responsible for the functional response during the light and the dark phase. The rank order of potency of agonists was dimethylphenylpiperazinium (DMPP) = cytisine > nicotine > carbachol and DMPP = nicotine = cytisine > carbachol, during the light and dark phase, respectively, due to an increase in apparent affinity for nicotine. Mecamylamine similarly blocked the DMPP response during the light and the dark phase, while the response to nicotine was more efficiently blocked during the light phase. In contrast, methyllycaconitine inhibited the nicotine-induced response only at 21:00 h. Since alpha7 nicotinic acetylcholine receptors (nAChRs) have low affinity for nicotine in binding assays, we suggest that a mixed population composed of alpha3ss4 - plus alpha7-bearing nAChR subtypes is present at night. This plasticity in receptor subtypes is probably driven by melatonin since nicotine-induced contraction in organs from animals sacrificed at 15:00 h and incubated with melatonin (100 pg/ml, 4 h) is not totally blocked by mecamylamine. Thus melatonin, by acting directly on the short adrenergic neurons that innervate the rat vas deferens, induces the appearance of the low affinity binding site, probably an alpha7 nAChR subtype. PMID- 10454763 TI - Effect of SX-3228, a selective ligand for the BZ1 receptor, on sleep and waking during the light-dark cycle in the rat. AB - The effects of the benzodiazepine1 (BZ1) receptor agonist SX-3228 were studied in rats (N = 12) implanted for chronic sleep procedures. Administration of 0.5, 1.0 and 2.5 mg/kg SX-3228, sc, to rats 1 h after the beginning of the light phase of the light-dark cycle induced a significant reduction of rapid-eye-movement sleep (REMS) during the third recording hour. Moreover, slow wave sleep (SWS) was increased during the fourth recording hour after the two largest doses of the compound. Administration of 0.5, 1.0 and 2.5 mg/kg SX-3228 one hour after the beginning of the dark period of the light-dark cycle caused a significant and maintained (6-h recording period) reduction of waking (W), whereas SWS and light sleep (LS) were increased. REMS values tended to increase during the entire recording period; however, the increase was statistically significant only for the 1.0 mg/kg dose during the first recording hour. In addition, a significant and dose-related increase of power density in the delta and the theta regions was found during nonREM sleep (LS and SWS) in the dark period. Our results indicate that SX-3228 is a potent hypnotic when given to the rat during the dark period of the light-dark cycle. Moreover, the sleep induced by SX-3228 during the dark phase closely resembles the physiological sleep of the rat. PMID- 10454765 TI - Structure and function of the cystic fibrosis transmembrane conductance regulator. AB - Cystic fibrosis (CF) is a lethal autosomal recessive genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR). Mutations in the CFTR gene may result in a defective processing of its protein and alter the function and regulation of this channel. Mutations are associated with different symptoms, including pancreatic insufficiency, bile duct obstruction, infertility in males, high sweat Cl-, intestinal obstruction, nasal polyp formation, chronic sinusitis, mucus dehydration, and chronic Pseudomonas aeruginosa and Staphylococcus aureus lung infection, responsible for 90% of the mortality of CF patients. The gene responsible for the cellular defect in CF was cloned in 1989 and its protein product CFTR is activated by an increase of intracellular cAMP. The CFTR contains two membrane domains, each with six transmembrane domain segments, two nucleotide-binding domains (NBDs), and a cytoplasmic domain. In this review we discuss the studies that have correlated the role of each CFTR domain in the protein function as a chloride channel and as a regulator of the outwardly rectifying Cl- channels (ORCCs). PMID- 10454764 TI - Relationship between the actions of atrial natriuretic peptide (ANP), guanylin and uroguanylin on the isolated kidney. AB - Guanylin and uroguanylin are peptides that bind to and activate guanylate cyclase C and control salt and water transport in many epithelia in vertebrates, mimicking the action of several heat-stable bacteria enterotoxins. In the kidney, both of them have well-documented natriuretic and kaliuretic effects. Since atrial natriuretic peptide (ANP) also has a natriuretic effect mediated by cGMP, experiments were designed in the isolated perfused rat kidney to identify possible synergisms between ANP, guanylin and uroguanylin. Inulin was added to the perfusate and glomerular filtration rate (GFR) was determined at 10-min intervals. Sodium was also determined. Electrolyte dynamics were measured by the clearance formula. Guanylin (0.5 microg/ml, N = 12) or uroguanylin (0.5 microg/ml, N = 9) was added to the system after 30 min of perfusion with ANP (0.1 ng/ml). The data were compared at 30-min intervals to a control (N = 12) perfused with modified Krebs-Hanseleit solution and to experiments using guanylin and uroguanylin at the same dose (0.5 microg/ml). After previous introduction of ANP in the system, guanylin promoted a reduction in fractional sodium transport (%TNa+, P<0.05) (from 78.46 +/- 0.86 to 64.62 +/- 1.92, 120 min). In contrast, ANP blocked uroguanylin-induced increase in urine flow (from 0.21 +/- 0.01 to 0.15 +/- 0.007 ml g-1 min-1, 120 min, P<0.05) and the reduction in fractional sodium transport (from 72.04 +/- 0. 86 to 85.19 +/- 1.48, %TNa+, at 120 min of perfusion, P<0.05). Thus, the synergism between ANP + guanylin and the antagonism between ANP + uroguanylin indicate the existence of different subtypes of receptors mediating the renal actions of guanylins. PMID- 10454766 TI - A simple RT-PCR-based strategy for screening connexin identity. AB - Vertebrate gap junctions are aggregates of transmembrane channels which are composed of connexin (Cx) proteins encoded by at least fourteen distinct genes in mammals. Since the same Cx type can be expressed in different tissues and more than one Cx type can be expressed by the same cell, the thorough identification of which connexin is in which cell type and how connexin expression changes after experimental manipulation has become quite laborious. Here we describe an efficient, rapid and simple method by which connexin type(s) can be identified in mammalian tissue and cultured cells using endonuclease cleavage of RT-PCR products generated from "multi primers" (sense primer, degenerate oligonucleotide corresponding to a region of the first extracellular domain; antisense primer, degenerate oligonucleotide complementary to the second extracellular domain) that amplify the cytoplasmic loop regions of all known connexins except Cx36. In addition, we provide sequence information on RT-PCR primers used in our laboratory to screen individual connexins and predictions of extension of the "multi primer" method to several human connexins. PMID- 10454767 TI - Effect of ryanodine on sinus node recovery time determined in vitro. AB - Evidence has indicated that the sarcoplasmic reticulum (SR) might be involved in the generation of spontaneous electrical activity in atrial pacemaker cells. We report the effect of disabling the SR with ryanodine (0.1 microM) on the sinus node recovery time (SNRT) measured in isolated right atria from 4-6-month-old male Wistar rats. Electrogram and isometric force were recorded at 36.5 degree C. Two methods for sinus node resetting were used: a) pulse: a single stimulus pulse interpolated at coupling intervals of 50, 65 or 80% of the regular spontaneous cycle length (RCL), and b) train: a 2-min train of pulses at intervals of 50, 65 or 80% of RCL. Corrected SNRT (cSNRT) was calculated as the difference between SNRT (first spontaneous cycle length after stimulation interruption) and RCL. Ryanodine only slightly increased RCL (<10%), but decreased developed force by 90%. When the pulse method was used, cSNRT ( approximately 40 ms), which represents intranodal/atrial conduction time, was independent of the coupling interval and unaffected by ryanodine. However, cSNRT obtained by the train method was significantly higher for shorter intervals between pulses, indicating the occurrence of overdrive suppression. In this case, ryanodine prolonged cSNRT in a rate-dependent fashion, with a greater effect at shorter intervals. These results indicate that: a) a functional SR, albeit important for force development, does not seem to play a major role in atrial automaticity in the rat; b) disruption of cell Ca2+ homeostasis by inhibition of SR function does not appear to affect conduction; however, it enhances overdrive-induced depression of sinusal automaticity. PMID- 10454768 TI - Inhibitory effect of high [Mg2+] on the vasopressin-stimulated hydroosmotic permeability of the isolated perfused cortical collecting duct. AB - High magnesium concentration inhibits the effect of arginine vasopressin (AVP) on smooth muscle contraction and platelet aggregation and also influences hepatocyte AVP receptor binding. The aim of this study was to determine the role of magnesium concentration [Mg2+] in AVP-stimulated water transport in the kidney collecting duct. The effect of low and high peritubular [Mg2+] on the AVP stimulated osmotic water permeability coefficient (Pf) was evaluated in the isolated perfused rabbit cortical collecting duct (CCD). Control tubules bathed and perfused with standard Ringer bicarbonate solution containing 1 mM Mg2+ presented a Pf of 223.9 +/- 27.2 microm/s. When Mg2+ was not added to the bathing solution, an increase in the AVP-stimulated Pf to 363.1 +/- 57.2 microm/s (P<0. 05) was observed. An elevation of Mg2+ to 5 mM resulted in a decrease in Pf to 202.9 +/- 12.6 microm/s (P<0.05). This decrease in the AVP-stimulated Pf at 5 mM Mg2+ persisted when the CCDs were returned to 1 mM Mg2+, Pf = 130.2 +/- 20.3 microm/s, and was not normalized by the addition of 8-[4-chlorophenylthio] adenosine 3', 5'-cyclic monophosphate, a cAMP analogue, to the preparation. These data indicate that magnesium may play a modulatory role in the action of AVP on CCD osmotic water permeability, as observed in other tissues. PMID- 10454772 TI - Seizures following renal transplantation in childhood. AB - Few studies have investigated the incidence of seizures following renal transplantation in childhood. The aim of this study was to determine this incidence and to identify risk factors. Retrospective casenote analysis was carried out on 119 transplants performed in 109 children over 10 years. Twenty one transplants (in 20 children) were complicated by seizures, the majority of which occurred in the first 55 days after transplantation. Seizures were more common in the 5- to 10-year-old age group (P=0.03), but were no more common in those with a prior history of seizure (P=0.69). Their aetiology was predominantly multifactorial; hypertension (n=15), fever/infection (n=4) and acute allograft rejection (n=6) were commonly identified risk factors; 2 were secondary to intracerebral pathology. Most seizures were short lived, required minimal therapy and had a good long-term neurological outcome. In conclusion, seizures are relatively common following paediatric renal transplantation. Parents are now routinely counselled of this risk. PMID- 10454773 TI - A familial syndrome due to Arg648Stop mutation in the X-linked renal chloride channel gene. AB - We describe a familial syndrome in two brothers who were investigated after the casual discovery of tubular proteinuria in their 1st month of life. During a follow-up of 20 and 11 years, respectively, the two children grew well and were asymptomatic, but developed the same biochemical abnormalities, i.e., tubular proteinuria and hyperphosphaturia, progressive decrease in serum phosphorus below the normal values for age, and an increase in serum 1,25-dihydroxyvitamin D levels over normal values. Moreover, hyperabsorptive hypercalciuria and systemic osteopenia developed and progressively worsened. In both children, at a different age, medullary nephrocalcinosis appeared. The oldest boy suffered a progressive decrease in urinary concentration ability and in glomerular filtration rate. Oral phosphate supplementation led to reversal of all biochemical abnormalities, with the exception of decreased phosphate tubular reabsorption and tubular proteinuria. With long-term phosphate supplementation, a normal bone mass was reached, while progression of nephrocalcinosis was arrested and impairment of renal function was slowed down. In a family study (siblings and parents), the only detectable abnormality was microglobinuria in the mother, thus suggesting a X-linked inheritance of this disorder. In the two probands a mutation within the renal chloride channel gene (CLCN5) was discovered. PMID- 10454774 TI - Intraperitoneal hypercoagulation and hypofibrinolysis is present in childhood peritonitis. AB - An increased rate of obstruction of peritoneal dialysis catheters is observed during peritonitis. Hypercoagulation and hypofibrinolysis may explain this increased occurrence. We studied plasminogen activator inhibitor type 1 antigen (PAI-1), tissue-type plasminogen activator antigen (t-PA), D-dimer (DD), plasmin alpha2-antiplasmin complexes (PAP), and thrombin-antithrombin III complexes (TAT) in 7 children with peritonitis (group A) and 12 children during stable peritoneal dialysis (group B). Albumin, beta2-microglobulin, IgG, and alpha2-macroglobulin were measured for baseline transperitoneal protein transport. After a dwell of 6 h with 1.36% Dianeal, dialysate and serum samples were collected. Dialysate to plasma ratios of all proteins were calculated. During peritonitis (group A) TAT was higher: 34.7 versus 22.0 (P=0.01). PAI-1 was increased in group A: 76.5 versus 22.9 (P=0.004). PAP was decreased during peritonitis (group A): 24.9 versus 39.3 (P=0.01). In group A, DD were decreased. 10.8 versus 26.7 (P=0.002). t-PA was similar in both groups (23.7 in group A vs. 27.7 in group B; P=0.26). In both groups TAT, PAI-1, t-PA, PAP, and DD were significantly higher than in baseline transperitoneal transport, suggesting intraperitoneal production. Hypercoagulability and hypofibrinolysis were present during peritonitis compared with the control situation. PMID- 10454775 TI - Chronic renal failure in pediatrics 1996. Chilean survey. AB - The Nephrology Branch of the Chilean Pediatric Society has greatly influenced the development of government health plans regarding the management and care of patients under 18 years with chronic renal failure (CRF). In order to assess the status of children with CRF in Chile up to 1996, a questionnaire was sent to all pediatric nephrologists in charge of those children. The total sample was of 227 patients under 18 years, giving a national prevalence of 42.5 and an incidence of 5.7 per million inhabitants; of these patients, 50.7% were male, 58.6% over 10 years and 15% younger than 5 years. The most frequent etiologies of CRF were: obstructive uropathy, 18.1%; hypo/dysplasia, 16.7%; reflux nephropathy, 16.7%; and glomerulopathies, 16.3%. Although 48% of patients were on conservative medical treatment, 42.2% of these were in end-stage renal disease, 22.9% were on dialysis, and 29.1% had undergone renal transplantation. Of the dialysis group, 75% were on peritoneal dialysis. Of the transplanted children, 78.8% had normal renal function, but 16.7% returned to dialysis. Three-year graft survival and patient survival were 68% and 94%, respectively. PMID- 10454776 TI - Extracellular fluid restoration in dehydration: a critique of rapid versus slow. AB - We compared current recommendations for treatment of severe dehydration by World Health Organization physicians and by the American Academy of Pediatrics Committee on Pediatric Gastroenterology with those in general textbooks of pediatrics, written mostly by pediatric nephrologists. The former recommend rapid (1- to 2-h) and generous intravenous restoration of extracellular fluid (ECF) volume followed by oral rehydration therapy (ORT) to replace potassium, current maintenance, and diarrheal losses--the rapid rehydration regimen. Oral feedings usually are resumed in 8-24 h. General textbooks of pediatrics usually recommend giving 20 ml/kg saline "to restore circulation," followed by the deficit therapy regimen to correct serum electrolyte abnormalities and replace remaining deficits of water, sodium, chloride, and potassium over 1-2 days. Mortality for hospitalized patients with dehydration treated with rapid rehydration was <3 per 1,000; no recent results are reported for patients treated by deficit therapy. The rapid rehydration regimen improves patient well being and restores perfusion, so that oral feedings are readily tolerated and renal function corrects serum electrolyte abnormalities in 6 h. Amounts of saline given correspond to amounts given for treating various forms of shock. Deficit therapy regimens provide less ECF restoration and are slower at restoring perfusion; tolerance for oral feedings is delayed. Two hundred pediatric nephrologists were surveyed, asking how they would treat a patient with severe dehydration and a patient with 40% burns. Only 30 of 200 responded; 29 used a deficit therapy regimen, with 20-40 ml/kg ECF replacement, while a majority rapidly and generously restored ECF volume in burn shock. We recommend that fluid therapy chapters should stop teaching deficit therapy for treating severe dehydration and instead teach the rapid rehydration regimen. PMID- 10454777 TI - Aggressive treatment of severe idiopathic focal segmental glomerulosclerosis. AB - When focal segmental glomerulosclerosis (FSGS) has reached the stage of chronic renal insufficiency, further progression is usually considered inevitable. African-American patients are believed to exhibit a particularly aggressive form of FSGS. We have treated five African-American patients, aged 11-18 years, with FSGS and reduced renal function using intensive intravenous methylprednisolone protocol, combined with chlorambucil in three cases. All patients had a pretreatment creatinine clearance of less than 50 ml/min per 1.73 m2. Three patients responded with normalization of creatinine clearance and serum albumin levels and had no or only minimal proteinuria at latest follow-up. One patient showed no improvement and one patient progressed to end-stage renal disease. These findings indicate, for the first time, that even severe FSGS may respond to aggressive methylprednisolone with or without alkylating agent treatment, and that African-American race does not preclude a favorable response. PMID- 10454778 TI - Treatment practices of FSGS among North American pediatric nephrologists. AB - A survey of North American pediatric nephrologists was conducted to assess the variability in the treatment of primary steroid-resistant focal segmental glomerulosclerosis (FSGS) of native kidneys. The most widely used immunosuppressive drug was cyclosporin A, with 73.9% using it often or sometimes. Only 44.3% used intravenous methylprednisolone combined with an alkylating agent at least sometimes; the use of methylprednisolone without cytotoxic drugs was slightly more common. Prolonged oral steroid therapy (>3 months) was used often or sometimes by 50.3%. The use of angiotensin converting enzyme inhibitors was very common, while lipid-lowering agents were rarely used. The variability in the treatment of FSGS most likely results from lack of evidence-based information and underscores the need for controlled pediatric multicenter treatment trials. PMID- 10454780 TI - Magnesium supplementation in Gitelman syndrome. AB - The metabolism of potassium and magnesium are closely linked (in situations where potassium and magnesium depletion coexist, magnesium restoration alone is sufficient to correct hypokalemia). Moreover, magnesium deficiency blunts the interplay between circulating calcium and the calciotropic hormones. Renal magnesium wasting, hypokalemia, alkalosis, hypocalciuria, and a tendency towards hypocalcemia characterize Gitelman syndrome. Plasma or intracellular potassium, circulating calcium, and calciotropic hormones were therefore investigated in eight patients (4 females, 4 males, aged 9-20 years) with Gitelman syndrome before and during oral supplementation with magnesium pyrrolidone carboxylate 30 mmol daily for 4 weeks. Magnesium supplementation significantly increased plasma and intracellular magnesium and plasma calcium, but failed to completely restore magnesium deficiency. In contrast, blood levels of parathyroid hormone and calcitriol and plasma and intracellular potassium were not modified following magnesium administration. PMID- 10454779 TI - Efficacy of amlodipine in pediatric patients with hypertension. AB - We designed a study to determine the efficacy and safety of amlodipine given once daily in the pediatric population. Twenty-one patients (mean age 13.1 years) with either essential (n=160) or renal (n=5) hypertension, and newly diagnosed (n=15) or poorly controlled or intolerant on existing antihypertensive therapy (n=6), were included. Patients received amlodipine once daily at a starting mean dose of 0.07+/-0.04 mg/kg per day. The total daily dose of amlodipine was increased 25% 50% every 5-7 days if the mean home blood pressure measurements (HBPM) were above the 95th percentile for age and gender. A baseline followed by a repeat 24-h ambulatory blood pressure monitor study (ABPM) was performed in 20 patients when the mean HBPM was below the 95th percentile goal. The mean titrated dose required to control BP was 0.29+/-0.11 mg/kg per day for those < 13 years, 0.16+/-0.11 mg/kg per day for those > or = 13 years, 0.23+/-0.14 mg/kg per day for essential, hypertension and 0.24+/-0.13 mg/kg per day for renal hypertension. The ABPM demonstrated that amlodipine provided effective BP control as primary therapy in 14 essential patients. Adverse effects included fatigue (n=6), headache (n=5), facial flushing (n=4), dizziness (n=3), edema (n=3), abdominal pain (n=3), chest pain (n=2), nausea (n=1), and vomiting (n=1). Quality of life appeared to improve during therapy. Amlodipine was an effective once daily antihypertensive agent with an acceptable safety profile. Higher doses of amlodipine were required for younger patients, and monotherapy was effective in patients with essential hypertension. PMID- 10454781 TI - Abnormal urinary acidification in infants with hydronephrosis. AB - Distal renal tubular abnormalities have been observed in patients with dilated urinary tract disorders. The present study was undertaken to look for patterns in urinary acidification in infants with varying degrees of hydronephrosis due to either reflux or obstruction and occurring as unilateral or bilateral disease. Three groups of infants (mean age 3.7+/-3.8 months) were studied prospectively. Groups IA and IB included patients with hydronephrosis who were acidotic and non acidotic, respectively. Group II served as controls and consisted of patients with diarrhea and secondary metabolic acidosis with no known renal disease. Serum electrolytes, creatinine, and urine pH were measured in all patients. Urinary titratable acidity, ammonium (NH4), and net acid excretion (NAE) were measured by the titrimetric method. Infants with hydronephrosis demonstrated lower urinary buffering capacity, reflected in low NAE in the face of acidosis. Deficiencies were noted in both titratable acid and NH4 excretion compared with control infants. Acidosis was as common in unilateral as in bilateral disease, regardless of severity score. These data confirm a defect in distal urinary acidification in infants with hydronephrosis, whether unilateral or bilateral. Immaturity and endogenous acid load may play a significant role in the manifestation of metabolic acidosis with unilateral disease. PMID- 10454782 TI - Effect of metabolic acidosis on branched-chain amino acids in uremia. AB - Fasting plasma concentrations of branched-chain amino acids (BCAA) valine, leucine, and isoleucine were measured in 20 young patients (aged 18+/-2 years) with end-stage renal disease just before initiation of dialysis and compared with 7 healthy controls (aged 19+/-1 years). Plasma valine, leucine, and isoleucine were all lower than control values (P<0.01 in all 3 cases). Plasma valine, but not leucine and isoleucine, correlated with venous pH (P<0.02). Plasma valine, leucine, or isoleucine did not correlate with blood urea nitrogen or serum creatinine. Seven patients (aged 18+/-1 years) on maintenance hemodialysis with metabolic acidosis were then studied before and after 2 weeks of oral sodium bicarbonate (NaHCO3) treatment to correct the acidosis. To control for the effect of additional sodium, they were also studied after 2 weeks of an equivalent amount of oral sodium chloride (NaCl). Oral NaHCO3 treatment led to significant increases in venous pH and serum bicarbonate concentrations, but no significant change in total and ionized calcium, phosphate, sodium, potassium, creatinine, blood urea nitrogen, and intact parathyroid hormone concentrations. Oral NaCl did not change any of the biochemical parameters. Fasting plasma concentrations of BCAA were measured. Before treatment of acidosis, uremic patients had low plasma concentrations of valine, leucine, and isoleucine compared with controls. Following 2 weeks of NaHCO3 treatment, there were significant increases in the plasma concentrations of valine and leucine (P<0.01), although the values did not normalize. There were no changes in plasma concentrations of valine and leucine following 2 weeks of NaCl. The plasma concentration of isoleucine was not different during baseline (acidotic) and treatment periods with NaHCO3 and NaCl. Thus treatment of metabolic acidosis ameliorated abnormalities in plasma concentrations of valine and leucine in patients with uremia on hemodialysis. PMID- 10454783 TI - Impact of plasmapheresis on disopyramide elimination. AB - The impact of plasmapheresis on the disposition of disopyramide was investigated in a 16-year-old female with systemic lupus erythematosus. Determination of total disopyramide plasma concentrations immediately prior to and following a 4-h plasmapheresis treatment revealed a significant reduction (i.e., 1.77 to 0.7 mg/l or approximately 60%). However, reassessment of the total serum concentration after 1.5 h (i.e., post equilibrium) revealed a rebounding of the value to 1.64 mg/l. Associated with this reduction in total serum levels was a decrease in the protein-bound fraction of disopyramide from 69.5% (pre treatment) to 48.6% (post treatment) that corresponded to a commensurate reduction in the concentration of alpha1-acid glycoprotein (i.e., 119 mg/dl pre treatment to 48.9 mg/dl post treatment). Despite these alterations in disopyramide concentrations, the procedure removed only 2.7% of the disopyramide dose and was not associated with the appearance of a cardiac dysrhythmia. PMID- 10454784 TI - Oxidative damage of erythrocyte membrane in nephrotic syndrome. AB - Erythrocytes are target cells for peroxidative damage. Abnormal susceptibility of erythrocyte lipids to peroxidation is believed to reflect a similar abnormality in other organs and tissues. The changes in erythrocyte lipid peroxidation [measured by malonyldialdehyde (MDA) concentration] and erythrocyte membrane cholesterol (EMC) and their correlation with plasma lipid changes were studied in 36 children with steroid-responsive minimal change nephrotic syndrome (MCNS) (16 in relapse, 20 in remission) and 30 matched healthy controls. Erythrocyte MDA levels were significantly higher in relapse [126.3+/-40.6 nmol/g hemoglobin (Hb)] compared with remission (101.2+/-21.3 nmol/g Hb, P<0.02) and in controls (95.4+/ 20.4 nmol/g Hb, P<0.001). Plasma MDA levels in relapse were also higher than in remission (4.26+/-1.19 nmol/ml vs. 3.16+/-1.18 nmol/ml, P<0.01), and in controls (2.49+/-0.86 nmol/ml, P<0.001). The EMC content changed significantly during remission (1.22+/-0.15 mg/10(10) cells in relapse, 1.09+/-0.19 mg/10(10) cells in remission, P<0.04). These results show an increased sensitivity of red cells to lipid peroxidation in patients with steroid-sensitive nephrotic syndrome without the development of renal failure and anemia. Lipid peroxidation of plasma lipids and erythrocyte membrane may be a primary phenomenon, but this should be confirmed by investigation of peroxidation of renal lipids. PMID- 10454785 TI - Acute plasmacytic interstitial nephritis in a child with Down syndrome. AB - A 7.5-year-old boy with Down syndrome presented in acute renal failure (ARF) needing dialysis. When 1.5 years old he had a neuroblastoma, was treated for 1 year with chemotherapy and radiotherapy, and off chemotherapy had since been in remission. Renal biopsy revealed an interstitial inflammation, principally of plasma cells with some lymphocytes and eosinophils. Immunofluorescence showed no deposition of immunoglobulins or complement (C3). The plasma cells were a mixture of kappa and lambda light chain-producing cells. The patient spontaneously improved a week after admission. Initial ultrasonography showed enlarged kidneys with loss of corticomedullary differentiation. We are unaware of a report of ARF in a child, resulting primarily from a polyclonal plasmacytic interstitial nephritis. The etiology remains unclear. PMID- 10454786 TI - Tubulointerstitial nephritis and uveitis in association with Epstein-Barr virus infection. AB - The case of a 13.5-year-old girl with acute tubulointerstitial nephritis and uveitis (TINU syndrome) is presented. The etiology of this rare syndrome, which in most cases involves female adolescents and usually regresses spontaneously, is still unknown. An infection-triggered pathological immune reaction has been considered to play a role in the pathogenesis of this disorder. Here we report for the first time the association of TINU syndrome and Epstein-Barr virus infection. PMID- 10454787 TI - Parvovirus glomerulonephritis in a patient with sickle cell disease. AB - A child with sickle cell disease developed glomerulonephritis 10 days following an aplastic crisis induced by human parvovirus B 19 infection. An initial kidney biopsy showed focal proliferative glomerulonephritis, and 1 year later was compatible with focal and segmental glomerulosclerosis. Renal tissue, tested by polymerase chain reaction, was positive for parvovirus, while the patient's blood was negative. For the first time a direct relationship has been demonstrated between parvovirus infection and glomerulonephritis. PMID- 10454788 TI - Severe hypertensive sequelae in a child with Seckel syndrome (bird-like dwarfism). AB - We report a 19-year-old male with Seckel syndrome (bird-like dwarfism) who presents with malignant hypertension associated with hypertensive nephrosclerosis, dilated cardiomyopathy, and a ruptured cerebral artery aneurysm. Although end-organ injury due to chronic hypertension occurs frequently in adults, no previous reports of renal insufficiency due to hypertension exist in children or adolescents. We speculate that this patient may have been particularly prone to hypertensive end-organ injury due to his extreme short stature. PMID- 10454789 TI - Development of the renal interstitium. AB - The kidney is derived from two tissue sources and develops through a series of mesenchymal-epithelial transitions and epithelial-mesenchymal interactions to form an epithelial tubular organ embedded in an interstitium derived from mesenchyme. The primary interstitium of the embryonic kidney undergoes significant differentiation to form the adult counterpart whose diverse cells have structural and functional characteristics that relate to their local milieu. Whether the adult interstitial cells retain the capacity to transform to other cell types and thus play a role in pathophysiological conditions appears more and more likely as the plasticity of cells becomes apparent. Besides forming the adult interstitium, the primary interstitium is active in metanephric development, with specific roles in nephron growth and collecting duct growth and arborization. Interruptions to the development of the interstitium or amelioration of its developmental capacity result in severely disrupted kidneys. The development of the renal interstitium is an essential component in the process of renal genesis. PMID- 10454790 TI - Vesicoureteral reflux in infants. AB - Grade 5 vesicoureteral reflux (VUR) is almost exclusively seen in male infants, and in one-third of cases occurring with a generalized small kidney with decreased renal function without a previous history of urinary tract infection. In females, however, high-grade reflux is rare and kidney damage almost always less severe and of the focal type, as in older children. Assessment of the bladder function with urodynamic and free voiding studies also indicates a difference between male and female reflux during infancy. Half of the males with dilating reflux initially have a hypercontractile urodynamic pattern indicating small functional capacity with high voiding pressures and often instability during filling. This pattern changes during the first couple of years to high capacity overdistended bladders, often with incomplete emptying. In females, hypercontractility is seldom seen, but bladder function is characterized by high capacity and there is an increase in residual urine from presentation. The pathogenesis of VUR has also been suggested to differ between the sexes. Transient anatomical obstruction during fetal life has been proposed as the cause of gross VUR in males. The spontaneous resolution rate of dilating infant VUR seems to be significantly higher than in older children. A resolution of 40% of grades 4 and 5 has been reported during the first couple of years in prenatally diagnosed cases, suggesting that antireflux surgery should be postponed until after the infant year. PMID- 10454791 TI - Clinical quiz. Tumoral calcinosis. PMID- 10454792 TI - The management of urinary tract infections. PMID- 10454793 TI - Adverse reactions to short-acting nifedipine in hypertensive crises. PMID- 10454794 TI - Pediatric reference values for cystatin C revisited. PMID- 10454795 TI - Parasite-induced bladder pathology. PMID- 10454796 TI - Proceedings of the 2nd International Symposium on Silk. Charlottesville, Virginia, USA. June 1998. PMID- 10454797 TI - Epiphora due to Kaposi's sarcoma of the nasolacrimal duct. PMID- 10454798 TI - The juxtaposition of ABL with BCR and risk for fusion may come at the time of BCR replication in late S-phase. PMID- 10454799 TI - Replication of hepatitis C virus in B lymphocytes (CD19+) PMID- 10454800 TI - Genetic heterogeneity of congenital dyserythropoietic anemia type I. PMID- 10454802 TI - Inter-strain variation in murine models of leukemia. PMID- 10454801 TI - Prognostic significance of c-ABL methylation in chronic myelogenous leukemia: still an open question. PMID- 10454803 TI - Sequence of Bcl-XL antisense oligonucleotides. PMID- 10454804 TI - Expression of laminin beta2 chain in normal human bone marrow. PMID- 10454805 TI - Proceedings of the 22nd International Symposium on Chromatography. Rome, Italy, 13-18 September 1998. Part III. PMID- 10454806 TI - Towards evidence based emergency medicine: best BETS from the Manchester Royal Infirmary. Admission of isolated sternal fracture for observation. PMID- 10454807 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal infirmary. Management of household electrical injury. PMID- 10454808 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Steroid delivery in croup. PMID- 10454809 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Haematoma block versus intravenous regional anaesthesia in Colles' fractures. PMID- 10454810 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Antibiotics and corneal abrasion. PMID- 10454811 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Lignocaine or bupivacaine for digital ring block. PMID- 10454812 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Steroids in lateral epicondylitis. PMID- 10454813 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Management of fractures of the neck of the fifth metacarpal. PMID- 10454814 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Follow up of a positive elbow fat sign. PMID- 10454815 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Steroids in De Quervain's tenosynovitis. PMID- 10454816 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Antibiotics after dog bite. PMID- 10454817 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Immobilisation after first anterior shoulder dislocation. PMID- 10454818 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Pelvic radiography in severe blunt trauma. PMID- 10454819 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal infirmary. The management of anterior epistaxis. PMID- 10454820 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Topical analgesia in corneal abrasions. PMID- 10454821 TI - Towards evidence based emergency medicine: best BETs from the Manchester Royal Infirmary. Wound cleaning methods. PMID- 10454822 TI - From Drosophila mutants to human disease. PMID- 10454823 TI - The relationship between congenital hypertrophy of the retinal pigment epithelium (CHRPE) and germline mutations in the adenomatous polyposis coli (APC) gene. PMID- 10454824 TI - Severe juvenile retinoschisis associated with a 33-bps deletion in XLRS1 gene. AB - X-linked juvenile retinoschisis is a form of vitreoretinal dystrophy that is characterized by foveal and peripheral splitting of the retinal nerve fiber layer. Pathognomonic of this disorder is a microcystic radiate appearance in the fovea. We encountered a 10 year-old, mildly retarded, Japanese boy, who exhibited a widely extended macular retinoschisis bilaterally. A break in the inner layer of the left eye mimicked a lamellar macular hole, which is a rare manifestation of the disease. Peripheral retinoschisis was absent. Only a few reports have described marked bilateral macular retinoschisis that involved entire posterior pole, while various other macular findings have been reported. This patient with a severe form of retinoschisis was found to harbor the deletion of 33 base pairs, including the boundary region of exon 3 and intron 3 in the XLRS1 gene. PMID- 10454825 TI - Endocrine cell transplantation and genetic engineering. Proceedings of an international symposium. Giessen, Germany, June 16-19, 1996. PMID- 10454827 TI - Differential effect of anti-TNF-alpha antibody on proinflammatory cytokine release by Kupffer cells following liver ischemia and reperfusion. AB - To study the role of tumor necrosis factor-alpha (TNF-alpha) for induction of the proinflammatory cytokine cascade after liver ischemia and reperfusion (I/R), rats were injected intraperitoneally with anti-TNF-alpha monoclonal antibodies (mAb) or placebo (IgG1) 30 min prior to global hepatic ischemia. Blood levels of TNF alpha, interleukin (IL)-1alpha and -6 were determined. In addition, Kupffer cells (KC) were harvested after 60 min of reperfusion and spontaneous cytokine release was measured. Sham-operated animals were used as controls. Levels of proinflammatory cytokines in serum and KC supernatants were detected using specific bioassays and ELISA. Liver I/R resulted in increased (p < .01) serum levels of TNF-alpha, IL-1alpha, and IL-6, which was associated with an enhanced (p < .05) release of these cytokines by KC. In vivo pretreatment with anti-TNF alpha mAb led to complete neutralization of TNF-alpha serum levels and decreased (p < .01) IL-6 levels (-62%). Moreover, anti-TNF-alpha mAb markedly (p < .05) decreased the release of TNF-alpha (-69%) and IL-6 (-56%) by KC, while IL-1alpha was not affected. These data indicate that TNF-alpha produced early after liver I/R triggers both its own secretion as well as IL-6 release by KC during reperfusion while the release of IL-1alpha occurs independent from TNF-alpha. PMID- 10454826 TI - Tumor necrosis factor alpha regulates CXC chemokine receptor expression and function. AB - The alpha chemokine family is central to the participation of neutrophils in the acute inflammatory response. These substances interact with neutrophils through two cell surface receptors, CXCR-1 and CXCR-2 (formally known as IL-8R-1 and IL 8R-2). We investigated the possible regulatory effects of tumor necrosis factor alpha (TNFalpha) pretreatment on CXCR-1 and CXCR-2. To this end, we examined these receptors with flow cytometry, radioligand binding, Northern blot analyzes, calcium mobilization, and chemotaxis experiments on human neutrophils. In flow cytometry experiments, TNFalpha pretreatment substantially decreased cell surface CXCR-2 receptor levels but showed partial recovery at 120 min. On the other hand, CXCR-1 receptor levels had a sharp decline at 15 min and maintained that level to 120 min. Northern blot analyzes showed that mRNA levels of both IL-8 receptors were essentially unchanged after 45 min of TNFalpha pretreatment, but declined markedly following 2 h of pretreatment. Chemotaxis experiments on cells treated with TNFalpha for 5-120 min showed a substantial down-regulation of chemotaxis to IL-8 and GROalpha. This was noted to be much greater than the decline in cell surface receptors. Calcium mobilization experiments revealed minimal inhibition of the IL-8-induced increase in calcium after pretreatment with TNFalpha, but the response to NAP-2 was substantially inhibited. The data demonstrate differential regulation of the IL-8 receptor. PMID- 10454828 TI - Mediator-dependent secondary injury after unilateral blunt thoracic trauma. AB - The pathophysiologic sequence leading to respiratory failure after chest trauma can be an inevitable consequence of the primary injury or a secondary, mediator driven inflammatory process. To distinguish between these alternatives, a simple cross-transfusion experiment was performed. A captive bolt gun injured the chest of anesthetized pigs that were mechanically ventilated with FiO2 = .21, .50, or .50 plus indomethacin (5 mg/kg intravenous; 15 min before injury). Tube thoracostomy immediately followed. After 30 min, blood from these injured donors was transfused into three matched groups of naive recipients (n = 8, 6, and 4, respectively) for a 33% exchange transfusion. Two control groups received blood from uninjured donors with tube thoracostomies only (FiO2 = .21, n = 7; FiO2 = .50, n = 10). Within 15-30 min after transfusion, in recipients from injured donors versus controls, lung compliance was decreased 20%, stroke volume and cardiac output were decreased 50%, and pulmonary vascular resistance was increased >300% (all p < .05). These changes recovered to baseline within 60-90 min. The stable metabolite of thromboxane A2, thromboxane B2, increased >500% in plasma within 15 min and remained elevated for >120 min. All responses were similar at 21 % or 50% O2, which suggests that hypoxia per se is not a cause of mediator production. All responses were eliminated by indomethacin. By 24 h, histologic changes included atelectasis in 3/3 recipients from injured donors versus 0/3 recipients from uninjured donors. We conclude that 1) blunt chest trauma releases blood borne mediators, including prostanoids; 2) these mediators can cause secondary cardiopulmonary changes in naive recipients similar to those produced by chest trauma; 3) the progression to trauma-induced respiratory failure is multifactorial; 4) early pharmacologic intervention, rather than supportive care alone, may benefit some victims of severe chest trauma. PMID- 10454829 TI - Role of Kupffer cells and PMN leukocytes in hepatic and systemic oxidative stress in rats subjected to tourniquet shock. AB - Kupffer cells (KCs) have been implicated in leukocyte recruitment and microvascular dysfunction associated with liver inflammation. The overall objective of this study was to assess the role of KCs and polymorphonuclear (PMN) leukocytes on the oxidative stress elicited in the liver as a consequence of hind limb reperfusion in rats subjected to tourniquet shock, a shock model that differs from other models in that hepatic injury is a consequence of remote organ damage. Colloidal carbon clearance from blood and its incorporation into KCs demonstrate that these cells are activated after the 2 h hind limb reperfusion period and that they are responsible for the observed oxidative stress and for PMN leukocyte recruitment and activation. Liver oxidative stress in this model is evidenced by increased liver tissue GSSG/GSH ratio, thiobarbituric acid reactive substances (TBARS), an index of lipid peroxidation, myeloperoxidase (MPO) activity, an index of tissue-associated neutrophil accumulation, and a significant loss in total tissue superoxide dismutase (SOD) activity. Mean arterial blood pressure (MAP), as well as plasma levels of alanine aminotransferase (ALT), an index of hepatic tissue injury, total SOD activity, plasma levels of alpha-tocopherol and beta-carotene, and total plasma nitrite are also affected as a consequence of KC activation after the 2 h hind limb reperfusion period. Inhibition of KC activity by gadolinium chloride (GdCl3) reverted most of the above alterations to values that do no differ from those found in control animals. These results support the hypothesis that hepatic and systemic oxidative stress elicited by hind limb reperfusion in rats subjected to tourniquet shock is both KC and PMN leukocyte dependent. PMID- 10454830 TI - Macrophage TNF mRNA expression induced by LPS is regulated by sphingomyelin metabolites. AB - Metabolism of macrophage (MO) membrane phospholipids produces key mediators of inflammation and major second messengers that modulate inflammatory responses during sepsis. Sphingomyelin is a major class of phospholipid that releases ceramide and sphingosine. This study was designed to investigate the involvement of sphingomyelin metabolites in MO activation by lipopolysaccharide (LPS). Rabbit alveolar MO were obtained by bronchoalveolar lavage and exposed to C6-ceramide, a cell-permeable analogue of natural ceramide, or sphingosine in the presence of Escherichia coli LPS (100 ng/mL). Tumor necrosis factor (TNF) mRNA expression was measured by Northern blot assays. Total nuclear extract was harvested for the measurement of nuclear factor KB (NFkappaB) with electrophoretic mobility shift assays. MO TNF production was measured by L929 bioassays. C-6 ceramide did not have any effects on MO TNF production or TNF mRNA expression with or without LPS stimulation. Inhibition of ceramide metabolism with 1-phenyl-2-decanoylamino-3 morpholino-1-propanol (PDMP), or N-oleoyl-ethanolamine (NOE) also did not induce TNF mRNA or TNF production. In comparison, sphingosine inhibited TNF mRNA expression as well as TNF production of LPS-stimulated MO. LPS-induced MO NFkappaB activity was also reduced by sphingosine. Our data indicate that ceramide alone has no effect on macrophage activity, but its metabolite sphingosine down-regulates MO activation induced by LPS stimulation. Therefore, the sphingomyelin pathway is involved in the regulation of MO activation. PMID- 10454831 TI - Early up-regulation of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression in rats with hemorrhagic shock and resuscitation. AB - This study evaluated the effect of resuscitation fluids on intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Sprague Dawley rats (n = 36) were subjected to a 27 mL/kg hemorrhage over 5 min followed by a 1 h shock and 1 h resuscitation. Animals groups included: 1) cannulation only (Sham); 2) hemorrhage only (NR); 3) resuscitation with 1:1 shed blood (Blood); 4) resuscitation with 3:1 lactated Ringer's (81 mL/kg, 3LR+); 5) no hemorrhage but infusion with 3:1 lactated Ringer's (3LR); and 6) resuscitation with .36:1 hypertonic saline (7.5%, 9.7 mL/kg, HTS). At the end of resuscitation, the spleen and lung were harvested for detection of adhesion molecule mRNA and protein by RT-PCR and immunostaining. ICAM-1 and VCAM-1 expression exhibited the following pattern: 3LR+ > HTS approximate to 3LR > Blood approximate to NR approximate to Sham. VCAM-1 mRNA in the lung of the 3LR+ group was 2 or more times more than the groups of Sham, NR, Blood, and 3LR (p < .05). ICAM-1 and VCAM 1 mRNA in the spleen was significantly increased in the 3LR+ group compared with the groups of Sham, NR, and Blood (p < .05). Animals in the 3LR+ group showed enhanced staining for ICAM-1 in the pulmonary microvessels and in the marginal and trabecular areas of the spleen. Pulmonary edema and inflammatory cell infiltration were observed only in the 3LR+ group. In summary, resuscitation with LR following hemorrhagic shock induced immediate up-regulation of ICAM-1 and VCAM 1, which was associated with tissue injury. Thus, the type of resuscitation fluid used affected resuscitation injury. PMID- 10454832 TI - Interleukin-1 mediates hemodynamic dysfunction and release of eicosanoids and tumor necrosis factor during graded bacteremia. AB - The pathophysiologic events of sepsis mediated by interleukin-1 (IL-1) remain ill defined. The purpose of this study was to identify the circulatory derangements of which IL-1 was a necessary mediator and evaluate its interactions with tumor necrosis factor (TNF) and the eicosanoids during graded bacteremia. Eleven adult female swine were anesthetized, mechanically ventilated, and monitored with pulmonary artery catheters and arterial lines; they received intravenously either saline vehicle (septic control, n = 6) or human recombinant IL-1 receptor antagonist (IL-1ra, n = 5). The animals were then infused with Aeromonas hydrophila (10(9)/mL) for 4 h at rates gradually increased from .2 mL/kg/h to 4 mL/kg/h over 3 h, then sacrificed after 4 h. Mean arterial pressure (MAP), left ventricular stroke work index (LVSWI), and systemic vascular resistance index (SVRI) were recorded at baseline and hourly thereafter, and plasma 6-keto PGF1alpha (6-KETO), tumor necrosis factor-alpha (TNF) and leukotrienes B4(LTB4) and C4D4E4 (LTCDE), pg/mL, were measured by ELISA. MAP, LVSWI, arterial P(O2) all decreased in the septic control group to levels significantly below those of the IL-1 antagonist animals. Circulating 6-KETO, LTCDE, and TNF increased significantly in all septic animals. Plasma LTB, and TNF were reduced by IL-1 blockade, compared with septic controls. TxB2 was not affected by IL-1 inhibition. There were no intergroup differences in platelet aggregation, but the in vitro aggregation response decreased from baseline in septic controls to 54+/ 27% (p < .05). IL-1 is necessary to the development of systemic hypotension impaired LVSWI, and increased intravascular platelet aggregation during graded bacteremia. Conversely, IL-1 helps to maintain stroke volume and low SVRI in graded bacteremia, possibly through increased prostacyclin release. It may contribute to impaired pulmonary gas exchange and increased tissue oxygen demands. TNF release is stimulated in the presence of unopposed IL-1 and may be synergistic with it in the adverse hemodynamic effects of endogenous IL-1. IL-1 is required for increased leukotriene and prostacyclin levels in this model, but it is not involved in thromboxane release. Whether the lack of survival benefit from IL-1ra in human sepsis is due to these mixed cardiopulmonary and mediator effects, to species differences, or to timing of IL-1ra administration is not clear from the data. PMID- 10454833 TI - Stimulation of liver RNA and protein breakdown in endotoxemic rats: role of glucocorticoids. AB - In systemic or localized acute inflammation, liver ribosomal RNA (rRNA) and protein contents increase. We first determined whether changes in RNA, more specifically rRNA, and protein breakdown rates were involved in the accumulation of both types of macromolecules 24 h after induction of endotoxemia. Liver RNA and protein contents were enhanced by 35 and 19%, respectively, in the endotoxemic rats. RNA and protein degradation rates measured during in situ cyclic perfusions of the livers were significantly higher in the endotoxemic rats than in the controls (42 and 46%, respectively). In order to check that the stimulation of RNA and protein degradation corresponded to an activation of the hepatocyte autophagic pathway, the fractional cytoplasmic volume (FCV) of autophagosomes, digestive autophagic vacuoles and dense bodies was measured by morphometry in electron microscopy. The FCV of the sum of these lysosomal structures was significantly increased in the endotoxemic rats. We next tried to identify the factor(s) responsible for the high breakdown rates. The increase in macromolecular degradation did not result from reduced portal amino acid supply. The effects of dexamethasone, interleukin-6, interleukin-1beta, and tumor necrosis factor alpha on RNA degradation were then investigated in primary cultures of hepatocytes isolated from control rats. Only dexamethasone stimulated RNA breakdown. Finally, pretreatment of endotoxemic rats with RU 38486, a glucocorticoid receptor antagonist, completely abolished the stimulation of RNA degradation observed in the sham-gavaged LPS-treated rats. Our data suggest an important role of glucocorticoids in the high levels of RNA and protein breakdown in endotoxemic rats. PMID- 10454834 TI - Enhanced prostanoid-mediated vasorelaxation in pulmonary arteries isolated during experimental endotoxemia. AB - Endotoxemia secondary to gram-negative sepsis has been shown to inhibit endothelium-dependent vasomotion in numerous vascular beds, including guinea pig aortae and coronary arteries. We tested the hypothesis that in vivo endotoxin impairs endothelium-dependent nitric oxide-mediated relaxation responses of pulmonary arteries isolated from guinea pigs given intraperitoneal injections of Escherichia coli endotoxin lipopolysaccharide (LPS) or saline (control) 16 h before sacrifice. Pulmonary rings from the main artery and primary branches were isolated and studied in vitro using conventional isometric techniques. Interestingly, endotoxemia resulted in enhanced pulmonary artery relaxation in response to the endothelium-dependent receptor agonists acetylcholine (10(-10) 10(-5) M) and adenosine diphosphate (ADP; 10(-9) -10(-5) M), as compared with control responses (p < .05). Nitric oxide synthase inhibitors N-monomethyl-L arginine (300 microM) and N-nitro-L-arginine methyl ester (100 microm) reduced acetylcholine- and adenosine diphosphate-mediated relaxation in both groups (p < .05); however, vasodilation responses in arteries from LPS animals remained enhanced relative to those of control arteries. In contrast to nitric oxide synthase inhibitors, the cyclooxygenase inhibitor indomethacin markedly inhibited acetylcholine- and adenosine diphosphate-mediated relaxation responses of pulmonary arteries isolated from LPS-treated animals (p < .05) but not control arteries; indomethacin effectively reversed LPS-induced enhanced vasodilation of pulmonary arteries. Relaxation responses to the receptor-independent calcium ionophore (A23187) and to the direct smooth muscle vasodilator sodium nitroprusside (+ N-nitro-L-arginine methyl ester) were not significantly altered by LPS treatment (p > .05). These data suggest that in pulmonary arteries, unlike aortae and coronary arteries isolated from the same model, in vivo LPS enhances agonist-mediated endothelium-dependent vasodilation responses to acetylcholine and adenosine diphosphate. Underlying mechanisms appear to involve increased dependency upon vasodilator prostanoids and decreased dependency on nitric oxide synthesis/release for LPS-induced alterations in pulmonary relaxation responses. PMID- 10454836 TI - Current literature. PMID- 10454835 TI - The role of cyclooxygenase in regulation of vasomotor tone during endotoxemia. PMID- 10454837 TI - Current literature. PMID- 10454838 TI - Current literature. PMID- 10454839 TI - Current literature. PMID- 10454840 TI - Genes and the heart: a quest for new therapeutic strategies. Cambridge Healthtech Institute's First Cardiovascular Genomics Conference. Orlando, FL, USA, 11-12 January 1999. PMID- 10454841 TI - Negative air ions and reactive oxygen species. Comment on A.V. Peskin's comment "Concerning the role of active oxygen species as regulators". PMID- 10454842 TI - Drug interactions: Who warns the patient. PMID- 10454843 TI - Waiting-list Web site "inaccurate" and "misleading," BC doctors complain. PMID- 10454844 TI - Waiting-list worries cause Calgary MDs to prepare letter for patients. PMID- 10454845 TI - Proceedings of the 3rd International Meeting on Esterases Reacting with Organophosphorus Compounds. Dubrovnik, Croatia, 15-18 April, 1998. PMID- 10454847 TI - Fitness helps fend off diabetes. PMID- 10454846 TI - Kikuchi's disease with multisystemic involvement and adverse reaction to drugs. AB - Kikuchi's disease (KD), or histiocytic necrotizing lymphadenitis, was initially described in Japan in 1972. In the following years, several series of cases involving patients of different ages, races, and geographic origins were reported, but pediatric reports have been rare. The etiology of KD is unknown, although a viral or autoimmune hypothesis has been suggested. The most frequent clinical manifestation consists of local or generalized adenopathy, although in some cases, it is associated with more general symptoms, multiorganic involvement, and diverse analytic changes (leukopenia, elevated erythrocyte sedimentation rate, and C-reactive protein, as well as an increase of transaminases and serum lactic dehydrogenase). Diagnosis is based on characteristic pathologic findings that permit differentiation of this disease from lymphoma, systemic lupus erythematosus, and infectious lymphadenopathies. We present here the case of a 14-year-old boy who presented with severe systemic manifestations and transient fulminant hepatic failure in response to treatment with antituberculosis drugs. Kikuchi's disease, lymphadenitis, liver failure, antituberculosis drugs. PMID- 10454849 TI - Home hepatitis C test. PMID- 10454848 TI - Weak bones and heart-attack risk. PMID- 10454850 TI - By the way, doctor. I am 70 years old and have taken estrogen (Premarin 1.25 mg) for 14 years. Recently, my bone density test showed osteopenia, which my doctor says is the beginning of osteoporosis. Why would I still get osteoporosis after taking large doses of estrogen for so long? PMID- 10454851 TI - By the way, doctor. I found your article on Alzheimer's disease (March 99) very interesting. Could you please tell me the difference between "Alzheimer's" and "senile dementia." Are the two interchangeable? PMID- 10454853 TI - Urological Research Society annual scientific meeting. London, United Kingdom, 8 January 1999. Abstracts. PMID- 10454852 TI - Technique to remove interference caused by radio-opaque agents in clinical capillary zone electrophoresis. PMID- 10454854 TI - Appropriateness of referral to urologists: can it be defined for symptoms of benign prostatic obstruction and used as a quality measure? PMID- 10454855 TI - Circumcision. PMID- 10454856 TI - Lower moiety heminephroureterectomy in duplex refluxing kidneys: the accuracy of isotopic scintigraphy in functional assessment. PMID- 10454857 TI - The Amplatz sheath in the female urethra: a safe and effective approach for cystolitholapaxy. PMID- 10454858 TI - Techniques of endopyelotomy. PMID- 10454859 TI - Predictive preventive medicine for hypertensive cardiovascular disease. Proceedings of satellite symposium to WHO Forum '97. Kyoto, Japan, October 1997. PMID- 10454860 TI - The ten most commonly asked questions about sildenafil (Viagra). PMID- 10454861 TI - Percutaneous balloon mitral valvuloplasty: long-term follow-up. PMID- 10454862 TI - Proceedings of the 41st Annual Thomas L. Petty Aspen Lung Conference: Acute Lung Injury. PMID- 10454863 TI - Family doctors and patients: is effective nutrition interaction possible? Proceedings of an international workshop. Heelsum, the Netherlands, December 14 16, 1998. PMID- 10454864 TI - 31st European Pancreatic Club Meeting. Luneburg, Germany, July 28-31, 1999. Abstracts. PMID- 10454865 TI - Early postoperative feeding following major abdominal gynecologic surgery reduced the length of hospitalization. PMID- 10454866 TI - [Special issue on cardiac rehabilitation dedicated in memory of Professor Richard Rost]. PMID- 10454867 TI - Standardization of Growth Hormone Measurement, Evidence-Based Medicine. Proceedings of the 3rd KIGS/KIMS Expert Meeting on Growth and Growth Disorders. Sorrento, Italy, November 19-20, 1998. PMID- 10454868 TI - Mapping of the human NBC3 (SLC4A7) gene to chromosome 3p22. PMID- 10454869 TI - Immunological Approaches to Tumor Therapy. Proceedings of the 4th International Meeting on Idiotypic Network and Tumor Therapy by Gene Therapy and Drug Targeting. Bonn, Germany, March 6-7, 1998. PMID- 10454870 TI - The process of ageing and consequences for the vitamin requirement. Proceedings of a conference. Bonn, Germany, May 14-15, 1999. PMID- 10454871 TI - Cerebral metabolism during vegetative state and after recovery to consciousness. PMID- 10454872 TI - Electrical inexcitability of nerves and muscles in severe infantile spinal muscular atrophy. PMID- 10454873 TI - Acute overdosage and intoxication with carbidopa/levodopa can be detected in the subacute stage by measurement of 3-o-methyldopa. PMID- 10454874 TI - The use of olanzapine for movement disorder in Huntington's disease: a first case report. PMID- 10454875 TI - Transient hiccups after posteroventral pallidotomy for Parkinson's disease. PMID- 10454876 TI - Psychological adjustment and self reported coping in stroke survivors with and without emotionalism. PMID- 10454877 TI - Paraneoplastic stiff limb syndrome. PMID- 10454878 TI - Tetrodotoxin intoxication in a uraemic patient. PMID- 10454879 TI - Relation between critical illness polyneuropathy and axonal Guillain-Barre syndrome. PMID- 10454880 TI - Repetitive transcranial magnetic stimulation in the treatment of chronic negative schizophrenia: a pilot study. PMID- 10454881 TI - Lemuel Dary Wright (March 1, 1913-May 12, 1995). PMID- 10454883 TI - The mental health of juvenile offenders. PMID- 10454882 TI - Synthesis, structural studies and pharmacological activity of novel bicyclic compounds derived from 3,4-dihydropyridones: 4-aryl-7,7-dimethyl-2,5-dioxo 1,2,3,4,5,6,7,8-octahydroquinolines. AB - The purpose of this research is to characterize the possible vascular selectivity of a series of novel bicyclic compounds derived from 3,4-dihydropyridones. We describe the synthesis, structural study by X-ray analysis and quantum chemical calculations at semiempirical (AMI) and ab initio (HF/321G) levels and pharmacological activity of these 4-aryl-7,7-dimethyl-2,5-dioxo-1,2,3,4,5,6,7,8 octahydroquinolines. In addition, the more favoured conformation for compounds 4a c in solution was determined from the calculated and experimental proton coupling constants. We report the first computational study on the structure of octahydroquinolines. The results of ab initio (HF/3-21G) and semiempirical molecular orbital calculations (AMI) are compared with the data obtained by X-ray crystallographic study for 4a. PMID- 10454884 TI - [Prevention of spontaneous bacterial Peritonitis. Comment on the contribution by J. Zundler, J. C. Bode: Spontaneous bacterial peritonitis]. PMID- 10454885 TI - [Squamous cell carcinoma of finger]. PMID- 10454886 TI - [Glucogonoma syndrome]. PMID- 10454887 TI - [Phakomatosis pigmentovascularis (type IIIa)]. PMID- 10454888 TI - [Cutis verticis gyrata]. PMID- 10454889 TI - [Porokeratosis linearis]. PMID- 10454890 TI - [Disseminated skin abnormalities due to gout]. PMID- 10454891 TI - [Widely disseminated ectopic acne]. PMID- 10454892 TI - [Data on the bronchoscopic treatment of pulmonary abscesses]. PMID- 10454893 TI - Universal newborn hearing screening: should we leap before we look? PMID- 10454894 TI - Universal newborn hearing screening: should we leap before we look? PMID- 10454895 TI - A community-based randomized trial encouraging sun protection for children. PMID- 10454896 TI - Index of suspicion. Case 2. Fracture of the scaphoid bone. PMID- 10454897 TI - Index of suspicion. Case 3. Kawasaki disease. PMID- 10454898 TI - Current awareness in prenatal diagnosis. PMID- 10454899 TI - [In vitro bactericidal action of dissolved ozone on different strains of Mycobacterium tuberculosis]. AB - The mycobacterium tuberculosis (MBT) strains H37 Rv (USA and Czechoslovakia), Erdman, and multiresistant clinical strain 2255 having different milliliter levels of microbes were kept in the soluble ozone at the concentrations of 1-4 micrograms/ml for 30 and 60 minutes. The soluble ozone-treated strains were cultured in the Finn-I medium. The soluble ozone at the concentration was found to produce bactericidal and bacteriostatic effects on MBT, including multiresistant strains. The MBT strains resistant to antituberculosis agents were also more resistant to soluble ozone; however, longer exposure to the "therapeutical" concentrations of soluble ozone proved to damage multiresistant strains. PMID- 10454900 TI - In vitro anti-Helicobacter pylori activity of some flavonoids and their metabolites. AB - The inhibitory effect of flavonoids and phenolic acids, which could be transformed from flavonoids by human intestinal microflora, on the growth of Helicobacter pylori (HP) was investigated. Ponciretin, hesperetin, naringenin and diosmetin were active against HP. Among them, ponciretin was the most potent and its MIC was 10-20 micrograms/ml. However, these active compounds against HP did nearly not inhibit the urease activity of HP. PMID- 10454902 TI - The Electronic Plant Gene Register. PMID- 10454901 TI - Evaluation of the antimicrobial potency of tannins and related compounds using the microdilution broth method. AB - The antimicrobial activity of a total of 27 tannins and related compounds was evaluated against 8 microorganisms, including 2 Gram-positive (Bacillus subtilis, Staphylococcus aureus), 4 Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis), and 2 yeasts (Candida albicans, Cryptococcus neoformans). The compounds tested were generally found to possess only weak to moderate antibacterial, but fairly high anticryptococcal activities. Attention is given to structure-activity relationships with emphasis on simple galloyl esters, hydrolyzable tannins and proanthcyanidins among this class of secondary products. PMID- 10454903 TI - 9th Meeting of the European Neuropeptide Club. Ferrera, Italy, May 12-15, 1999. Abstracts. PMID- 10454904 TI - Autoantibodies and autoimmunity. Proceedings of an international conference. Oslo, Norway, June 24-28, 1999. Abstracts. PMID- 10454905 TI - Quantification of polycyclic aromatic hydrocarbons in soil at McMurdo Station, Antarctica. AB - Polycyclic aromatic hydrocarbons (PAHs) are a group of compounds that have attracted much attention over the past several years. The United States Environmental Protection Agency (USEPA) has identified numerous PAHs as known or probable human carcinogens. The quantity of PAHs in the environment has dramatically increased, with the majority emitted from fossil fuel combustion sources. Surface soil samples were collected at McMurdo Station, Antarctica (77 degrees 51S, 166 degrees 41E), during peak summer activity and analyzed for PAHs. PAHS were detected at several locations, with maximum concentrations for naphthalene, acenaphthene, acenaphthylene, and fluoranthene at 27,000, 17,800, 15,700 and 13,300 mg/kg, respectively. Results suggest anthropogenic activities may be contributing to increased levels of PAHs present in McMurdo soils. PMID- 10454906 TI - Mercury and other trace metals (Ag, Cr, Co, and Ni) in soft tissue and byssus of Mytilus edulis from the east coast of Kyushu Island, Japan. AB - Concentrations of Hg and other metals such as Ag, Cr, Co and Ni in soft tissues and byssi of the blue mussel (Mytilus edulis) from three sites along the east coast of Kyushu Island, Japan, were determined by a cold-vapor technique. Large inter-regional differences in metal concentrations in both soft tissues and byssi were observed. The tissue concentrations of Hg were significantly greater in M. edulis from the most contaminated locations (Saganoseki) compared to those from a non-industrialized area (Urashiro). It is evident that, compared to the soft tissue, byssus is a more sensitive biomonitor for Hg, Cr, Co and Ni. From the data obtained in the present study evidently results that the soft tissue and especially byssi of M. edulis appear to be good bioindicators for identification of coastal areas exposed to Hg contaminants. PMID- 10454907 TI - Report of new hepatitis virus has researchers intrigued and upset. PMID- 10454908 TI - A trigger of natural (and other) killers. PMID- 10454909 TI - Kassirer forced out at New England Journal. PMID- 10454910 TI - NIH to help fund big physics facilities. PMID- 10454911 TI - Stem cells as potential nerve therapy. PMID- 10454912 TI - Keck gives $110 million for USC initiatives. PMID- 10454913 TI - Gaining new insight into the molecular basis of evolution. PMID- 10454914 TI - AIDS vaccines. Glimmerings of hope from the bottom of the well. PMID- 10454915 TI - Science and "truth". PMID- 10454916 TI - Science and "truth". PMID- 10454917 TI - Science and "truth". PMID- 10454918 TI - Science and "truth". PMID- 10454919 TI - Retinoblastoma treatment. PMID- 10454920 TI - Asking the right questions. PMID- 10454921 TI - Proposed changes for NIH's Center for Scientific Review. Panel on Scientific Boundaries for Review. Center for Scientific Review Advisory Committee, National Institutes of Health. PMID- 10454922 TI - Perspectives: neuroscience. Is art lawful? PMID- 10454923 TI - Lentiviral vectors--the promise of gene therapy within reach? PMID- 10454925 TI - DOE builds a web site for the physical sciences. PMID- 10454924 TI - E-biomed morphs to E-biosci, focus shifts to reviewed papers. PMID- 10454926 TI - New clues to whooping cough pathology. PMID- 10454927 TI - Gene linked to faulty cholesterol transport. PMID- 10454928 TI - "Macho total synthesis". PMID- 10454929 TI - "Macho total synthesis". PMID- 10454930 TI - Chimp cultural diversity. PMID- 10454931 TI - Chimp cultural diversity. PMID- 10454932 TI - DNA discovery. PMID- 10454933 TI - Mitochondrial recombination? (continued) PMID- 10454934 TI - Protein crystallization at NASA: well grounded. PMID- 10454935 TI - Close encounters--an artist shows that size affects shape. PMID- 10454936 TI - The treatment of patients with autoimmune thrombocytopenia with intravenous IgG anti-D. PMID- 10454937 TI - [Aspects of cost effectiveness in therapy of acute leg/pelvic vein thrombosis]. AB - Out-patient or home treatment of acute proximal deep venous thrombosis (DVT) has not made its way yet. In this article, we review the data published on this form of management with regard to effectiveness and costs. We find that more than 80% of patients with DVT need not be admitted to the hospital and that the rate of secondary hospitalization is < 2%. Home treatment requires professional assistance in 15 to 74% of cases, while out-patient management with elastic leg compression and deliberate ambulation goes completely without it. The effect on objectivity assessed clinical symptoms, absence from work and patients' acceptance is significantly better and the costs are 2.5- to 3.2-fold lower with home or outpatient treatment than with treatment in the hospital. However, we found outpatient treatment associated with a reduced general well-being resulting from a probably frightening information which mentioned the possibility of experiencing an eventually fatal pulmonary embolism despite treatment. Management with deliberate ambulation in compression hosiery lead to a slower relief of subjective leg symptoms than initial immobilisation. We conclude that outpatient treatment of DVT is highly cost-effective. The problems still encountered are of a psychological nature and also have to do with treatment by leg compression and ambulation. These measures have to be optimized. PMID- 10454938 TI - Proceedings of the Helicobacter pylori Symposium. New Haven, Connecticut, USA. March 1-2, 1997. PMID- 10454939 TI - Photo page. Infinite worm. PMID- 10454940 TI - Educational update: "part-time" teachers in orthodontics programs. PMID- 10454941 TI - Product clarification. PMID- 10454942 TI - Alcohol-Induced Hepatic Fibrosis: Mechanisms. Proceedings of a satellite symposium. Hilton Head Island, South Carolina, USA. June 1998. PMID- 10454943 TI - Notice of withdrawal. PMID- 10454944 TI - Improving adherence to dementia guidelines through education and opinion leaders. A randomized, controlled trial. AB - BACKGROUND: Educational methods that encourage physicians to adopt practice guidelines are needed. OBJECTIVE: To evaluate an educational strategy to increase neurologists' adherence to specialty society-endorsed practice recommendations. DESIGN: Randomized, controlled trial. SETTING: Six urban regions in New York State. PARTICIPANTS: 417 neurologists. INTERVENTION: The educational strategy promoted six recommendations for evaluation and management of dementia. It included a mailed American Academy of Neurology continuing medical education course, practice-based tools, an interactive evidence-based American Academy of Neurology-sponsored seminar led by local opinion leaders, and follow-up mailings. MEASUREMENTS: Neurologists' adherence to guidelines was measured by using detailed clinical scenarios mailed to a baseline group 3 months before the intervention and to intervention and control groups 6 months after the intervention. In one region, patients' medical records were reviewed to determine concordance between neurologists' scenario responses and their actual care. RESULTS: Compared with neurologists in the baseline and control groups, neurologists in the intervention group were more adherent to three of the six recommendations: neuroimaging for patients with dementia only when certain criteria are present (odds ratio, 4.1 [95% CI, 1.9 to 8.9]), referral of all patients with dementia and their families to the Alzheimer's Association (odds ratio, 2.8 [CI, 1.7 to 4.8]), and encouragement of all patients and their families to enroll in the Alzheimer's Association Safe Return Program (odds ratio, 10.8 [CI, 3.5 to 33.2]). For the other three recommendations, adherence did not differ between the intervention and the nonintervention groups. Agreement between scenario responses and actual care ranged from 27% to 99% for the six recommendations and was 95% or more for three of the recommendations. CONCLUSION: A multifaceted educational program can improve physician adoption of practice guidelines. PMID- 10454945 TI - Early diagnosis and treatment of pancreatic dysplasia in patients with a family history of pancreatic cancer. AB - BACKGROUND: Pancreatic cancer, the fourth most common cause of cancer death in the United States, is hereditary in an estimated 10% of cases. Surveillance of patients with a familial predisposition for pancreatic cancer has not been systematically evaluated. OBJECTIVE: To develop a surveillance program that can identify and treat patients who have precancerous conditions of the pancreas and a family history of pancreatic cancer. DESIGN: Prospective cohort study. SETTING: University medical center. PATIENTS: 14 patients from three kindreds with a history of pancreatic cancer. INTERVENTIONS: Endoscopic ultrasonography, endoscopic retrograde cholangiopancreatography (ERCP), spiral computed tomography, and serum carcinoembryonic antigen and CA19-9 analysis were performed in all patients. Four affected patients were tested for the K-ras mutation. MAIN OUTCOME MEASUREMENT: Pancreatic dysplasia was determined by histologic evaluation. RESULTS: Seven of the 14 patients were believed to have dysplasia on the basis of clinical history and abnormalities on endoscopic ultrasonography and ERCP and were referred for pancreatectomy. All 7 patients had histologic evidence of dysplasia in pancreatectomy specimens. Findings on endoscopic ultrasonography were subtle, nonspecific, and similar to those seen in patients with chronic pancreatitis. Findings on ERCP ranged from mild and focal side-branch duct irregularities and small sacculations to main-duct strictures and grapelike clusters of saccules. Some of these changes are typical of chronic pancreatitis, but others are more distinctive. Spiral computed tomography and serum tumor markers had low sensitivity in the detection of pancreatic dysplasia. Analysis for the K-ras mutation yielded positive results in 3 of 4 patients with dysplasia. CONCLUSIONS: Thorough screening of patients with a family history of pancreatic cancer is feasible. Clinical data combined with imaging studies (endoscopic ultrasonography and ERCP) can be used to identify high-risk patients who have dysplasia. The role of molecular genetic testing is uncertain at this time. PMID- 10454946 TI - Plasma HIV viral load in patients with hemophilia and late-stage HIV disease: a measure of current immune suppression. Multicenter Hemophilia Cohort Study. AB - BACKGROUND: For patients infected with HIV, plasma HIV viral load in early disease predicts long-term prognosis. However, the implications of viral load measurements late in HIV disease are uncertain. OBJECTIVE: To evaluate the relation between plasma HIV viral load and subsequent risk for disease progression in patients with late-stage HIV disease. DESIGN: Retrospective cohort study. SETTING: 16 treatment centers for patients with hemophilia. PATIENTS: 389 patients with hemophilia and late-stage HIV disease (CD4 count < 200 cells/mm3). MEASUREMENTS: Plasma HIV viral load was measured at baseline. Patients were followed for AIDS-related illnesses (primary outcome) and, specifically, Pneumocystis carinii pneumonia (secondary outcome). RESULTS: HIV viral load strongly predicted AIDS-related illness. For patients with viral loads less than 4.00 log10 copies/mL, the 1-year actuarial risk was 0% and the 5-year risk was 25%. For patients with viral loads of at least 6.00 log10 copies/mL, the 1-year actuarial risk was 42% and the 5-year risk was 78%. A linear relation existed between viral load and risk for AIDS-related illness (hazard ratio, 2.37 per 1og10 copies/mL; P < 0.001). In addition, viral load most strongly predicted risk for illness immediately after viral load testing; this predictive relation attenuated over time (P = 0.002). These findings changed little after adjustment for CD4 cell counts that were updated during follow-up. In the first year after viral load was measured, it predicted occurrence of P. carinii pneumonia (hazard ratio, 4.69 per 1og10 copies/mL; P < 0.001). CONCLUSIONS: In patients with hemophilia and late-stage HIV disease, viral load predicts disease progression independently of CD4 cell counts. Because viral load most strongly predicts progression immediately after load is measured, it seems to reflect the current level of immunosuppression. PMID- 10454947 TI - Patient-identified needs for hormone replacement therapy counseling: a qualitative study. AB - BACKGROUND: Numerous medical organizations, including the American College of Physicians, have recommended that women be counseled about postmenopausal hormone replacement therapy (HRT). Patients' perspectives on their counseling needs, however, have not been integrated into most counseling guides. OBJECTIVE: To use patient self-reports to identify needs for HRT counseling. DESIGN: Individual, in depth patient interviews. SETTING: Managed care organization. PATIENTS: 26 women who had received an initial prescription for HRT. MEASUREMENTS: Qualitative, consensus review of the content of interview transcripts. RESULTS: On average, women reported 15 factors (range, 6 to 24 factors) as critical to their decision making process. Although most women cited their physician's opinion (96%), reports in the media (81%), and experiences and opinions of friends (77%) as critical to their decisions about HRT, counseling recommendations address none of these concerns. CONCLUSIONS: Many women in a managed care organization who accepted a prescription for HRT identified counseling needs that are not included in widely used HRT guidelines. PMID- 10454948 TI - Infection-control measures reduce transmission of vancomycin-resistant enterococci in an endemic setting. AB - BACKGROUND: Vancomycin-resistant enterococci (VRE) are nosocomial pathogens in many U. S. hospitals. OBJECTIVE: To determine whether enhanced infection-control strategies reduce transmission of VRE in an endemic setting. DESIGN: Prospective cohort study. SETTING: Adult oncology inpatient unit. PATIENTS: 259 patients evaluated during use of enhanced infection-control strategies and 184 patients evaluated during use of standard infection-control practices. INTERVENTIONS: Patient surveillance cultures were taken, patients were assigned to geographic cohorts, nurses were assigned to patient cohorts, gowns and gloves were worn on room entry, compliance with infection-control procedures was monitored, patients were educated about VRE transmission, patients taking antimicrobial agents were evaluated by an infectious disease specialist, and environmental surveillance was performed. MEASUREMENTS: VRE infection rates, VRE colonization rates, and changes in antimicrobial use. RESULTS: During use of enhanced infection-control strategies, incidence of VRE bloodstream infections decreased significantly (0.45 patients per 1000 patient-days compared with 2.1 patients per 1000 patient-days; relative rate ratio, 0.22 [95% CI, 0.05 to 0.92]; P = 0.04), as did VRE colonization (10.3 patients per 1000 patient-days compared with 20.7 patients per 1000 patient-days; relative rate ratio, 0.5 [CI, 0.33 to 0.75]; P < 0.001). Use of all antimicrobial agents except clindamycin and amikacin was significantly reduced. CONCLUSION: Enhanced infection-control strategies reduced VRE transmission in an oncology unit in which VRE were endemic. PMID- 10454949 TI - Update in infectious diseases. PMID- 10454951 TI - Guidelines have their limits. PMID- 10454950 TI - Pharmacologic therapy for type 2 diabetes mellitus. AB - Type 2 diabetes mellitus is a chronic metabolic disorder that results from defects in both insulin secretion and insulin action. An elevated rate of basal hepatic glucose production in the presence of hyperinsulinemia is the primary cause of fasting hyperglycemia; after a meal, impaired suppression of hepatic glucose production by insulin and decreased insulin-mediated glucose uptake by muscle contribute almost equally to postprandial hyperglycemia. In the United States, five classes of oral agents, each of which works through a different mechanism of action, are currently available to improve glycemic control in patients with type 2 diabetes. The recently completed United Kingdom Prospective Diabetes Study (UKPDS) has shown that type 2 diabetes mellitus is a progressive disorder that can be treated initially with oral agent monotherapy but will eventually require the addition of other oral agents, and that in many patients, insulin therapy will be needed to achieve targeted glycemic levels. In the UKPDS, improved glycemic control, irrespective of the agent used (sulfonylureas, metformin, or insulin), decreased the incidence of microvascular complications (retinopathy, neuropathy, and nephropathy). This review examines the goals of antihyperglycemic therapy and reviews the mechanism of action, efficacy, nonglycemic benefits, cost, and safety profile of each of the five approved classes of oral agents. A rationale for the use of these oral agents as monotherapy, in combination with each other, and in combination with insulin is provided. PMID- 10454952 TI - On the road. PMID- 10454953 TI - Update in endocrinology. PMID- 10454954 TI - Compliance with handwashing. PMID- 10454955 TI - Epidemiology of Mycobacterium kansasii. PMID- 10454956 TI - Diagnosis of hemochromatosis. PMID- 10454957 TI - Oral thymic extract to treat hepatitis C. PMID- 10454958 TI - Oral thymic extract to treat hepatitis C. PMID- 10454959 TI - Fluoroquinolone resistance in Streptococcus pneumoniae. PMID- 10454960 TI - Reversal of protease inhibitor-related visceral abdominal fat accumulation with recombinant human growth hormone. PMID- 10454961 TI - Acute hepatitis A in patients with chronic hepatitis C. PMID- 10454962 TI - Polyclonal hyperglobulinemia and spurious hypophosphatemia. PMID- 10454963 TI - Nutrition and policy. 5: Who should teach patients about nutrition? PMID- 10454964 TI - Neuroscience 2000: a new era of discovery. PMID- 10454965 TI - Molecular Strategies in Biological Evolution. Conference proceedings. New York, New York, USA. PMID- 10454966 TI - Cell and molecular biology of pancreatic carcinoma. Proceedings of the Baltic Pancreas Meeting on Pancreatic Carcinoma from Bench to Bedside. Rostock Warnemunde, Germany, September 17-18, 1998. PMID- 10454967 TI - Perioperative nursing in the early 1900s. PMID- 10454968 TI - Myocardial laser revascularization. PMID- 10454969 TI - Streptokinase...never again. PMID- 10454970 TI - Stress echo: more method than madness. PMID- 10454972 TI - The careers of female cardiologists. PMID- 10454971 TI - Cost-effective treatment of acute coronary syndromes-IIB or not IIB? PMID- 10454973 TI - Homocysteine, vitamins and sex. PMID- 10454974 TI - Grading, staging and scoring of left-ventricular hypertrophy, left-ventricular dilatation, asymptomatic left-ventricular dysfunction and chronic heart failure. PMID- 10454975 TI - Homocysteine, vitamin status and risk of vascular disease; effects of gender and menopausal status. European COMAC Group. AB - BACKGROUND: Elevated plasma total homocysteine (tHcy) is a known risk factor for vascular disease. Gender, age, and circulating levels of folate, vitamins B(6)and B(12)affect tHcy levels. Objectives To study associations of gender and age with levels of plasma tHcy, and to examine the relationships of tHcy and circulating levels of folate, vitamins B(6)and B(12)with risk of vascular disease in men and women (pre- and post-menopausal). MATERIAL AND METHODS: In a multicentre case control study in Europe, 750 patients (544 men, 206 women) with documented vascular disease of the coronary, cerebral, or peripheral vessels and 800 control subjects (570 men, 230 women) were enrolled. Plasma tHcy levels (fasting and after methionine loading) and circulating levels of the vitamins were measured. Adjustment for age and centre was carried out for all statistical analyses, with additional adjustment for serum creatinine and vitamins for the tHcy comparisons between the sexes and between cases and controls. Risk analyses included adjustment for creatinine and traditional risk factors. Relationships between age, gender and tHcy were studied among control subjects only. RESULTS: Fasting tHcy levels were lower in women than in men. Levels of tHcy showed a positive association with age, for both sexes. In the post-menopausal age category, female post-methionine load tHcy levels surpassed levels of men. Elevation of tHcy (defined as >80th percentile of controls) appeared to be at least as strong a risk factor for vascular disease in women as in men, even before the menopause. For post-methionine load tHcy, there was a 40% stronger association with vascular disease in women than in men. In both sexes, but especially in pre-menopausal women, low circulating levels of vitamin B(6)conferred a two- to threefold increased risk of vascular disease, independent of tHcy. In men, but not in women, low (defined as <20th percentile of controls) circulating folate levels were associated with a 50% increased risk of vascular disease. CONCLUSIONS: Elevation of tHcy appears to be at least as strong a risk for vascular disease in women as men, even before the menopause. Our data indicate that associations of the various tHcy measurements (and the vitamins that determine them), with risks of vascular disease may differ between the sexes. The tHcy-independent relationship of vitamin B(6)with vascular disease indicates that it will be advisable to test the effects of vitamin B(6)in clinical trials. PMID- 10454976 TI - Humoral and cellular immune responses up to 7.5 years after administration of streptokinase for acute myocardial infarction. AB - AIMS: Administration of streptokinase results in an immunological response which may lead to increased risk of anaphylactic reaction or reduced thrombolytic efficacy on repeat administration. For these reasons current recommendations suggest that streptokinase should not be given up to 1 year after first administration. We sought to define the profile of both the circulating antibody and T-cell response to streptokinase in patients who had received streptokinase up to 7.5 years previously following acute myocardial infarction. METHODS: Neutralizing anti-streptokinase antibody and total anti-streptokinase IgG were measured in 219 patients who had suffered acute myocardial infarction between 12 and 90 months previously and had received streptokinase. T-cell response to streptokinase was assessed by in-vitro proliferation of peripheral blood mononuclear cells (n=234). Data on all parameters were available in 184 patients. Controls (n=22) had suffered acute myocardial infarction between 73 and 84 months previously but had not received thrombolytic therapy. RESULTS: Compared to controls, anti-streptokinase antibodies were elevated at all time periods from 12 to 90 months after streptokinase treatment. Total anti-streptokinase titres showed the expected correlation with neutralizing anti-streptokinase antibodies (P<0.0001). Peripheral blood mononuclear cells showed a vigorous in-vitro proliferative response to streptokinase 6 days after treatment (P=0.05 vs pre treatment), but this was not detectable at 6 weeks or subsequently. CONCLUSION: There is as yet no evidence of a time limit beyond which administration of streptokinase on a second occasion can be regarded as safe and likely to be effective. Measurement of neutralizing anti-streptokinase or total anti streptokinase IgG titre appear to provide equivalent information regarding the antibody status of a population. Further studies are required regarding the apparent lack of peripheral blood mononuclear cells responsiveness in patients previously exposed to streptokinase. PMID- 10454977 TI - Economic assessment of tirofiban in the management of acute coronary syndromes in the hospital setting: an analysis based on the PRISM PLUS trial. AB - AIMS: We analysed whether generalized use of tirofiban plus heparin and aspirin might save direct healthcare costs, as compared with heparin and aspirin alone, in patients with acute coronary ischaemic syndromes in Switzerland. METHODS AND RESULTS: We conducted an incremental cost-consequence analysis from the perspective of the admitting hospital for the period of the first 7 days. Costs were analysed for the management of refractory ischaemic conditions and myocardial infarctions, including incremental days on the general ward or intensive care unit, as well as necessary revascularization procedures, and expressed in Swiss francs (CHF) and European currency units (ECU). Drug costs were based on a loading dose of 0.4 micro x kg(-1) x min(-1)and a maintenance dose of 0.1 micro x kg(-1) x min(-1)for tirofiban at a cost of CHF 273.55 (ECU 166.50) per vial. Heparin was administered at a loading dose of 5000 U and a maintenance dose of 1000 U x h(-1). All calculations were standardized to 100 treated patients. The costs of managing ischaemic complications were based on typical practice patterns in Swiss hospitals. The incremental costs per patient of managing unstable angina patients with recurrent ischaemia or myocardial infarction were calculated as CHF 23 325 (ECU 14 198) and CHF 18 599 (ECU 11 321), respectively. The incremental drug costs amounted to CHF 82 065 (ECU 49 954). The additional use of tirofiban resulted in net savings of CHF 54 899 (ECU 33 418) per 100 patients, achieved through a reduction in the cost of treating refractory ischaemic conditions (CHF 79 306, ECU 48 275) and myocardial infarctions (CHF 57 658, ECU 35 097). CONCLUSION: Tirofiban is cost-saving in acute coronary ischaemic syndromes and improves the economics of managing these patients during the initial hospitalization. PMID- 10454978 TI - Madness and method in stress echo reading. AB - AIM: To assess whether 'eye education' through short-term, high-intensity joint reading sessions may improve diagnostic accuracy and inter-observer agreement among beginners. METHODS AND RESULTS: Seventeen cardiologists with absent to minimal (<100 studies performed) previous stress echo experience independently and blindly read 18 stress echo studies, nine at the beginning ('pre-training' set) and nine at the end ('post-training' set) of a 2 day stress echo school which included a joint reading session of 50 tapes. The two sets were balanced as far as type of stress and image quality. The 17 observers had an average accuracy score of 51+/-16.4 before and 64.3+/-8.7% after the training (P<0.005). Concordant (i.e. >14 readers giving the same response) interpretation occurred in three out of nine studies before and in eight out of nine studies after the training (33% vs 88%, P<0.01). Kappa values went from 0.14 (poor) before to 0.39 (fair, close to moderate) after the training. CONCLUSION: Short-term, high intensity dedicated training in stress echo, with joint reading sessions and consensus development of reading criteria significantly increased accuracy and markedly reduced the inter-observer variability in the reading of stress echoes by beginners. If there is a Shakespearean madness in stress echo reading, 'yet there is a method in't' (Hamlet, II, II, 205-206). PMID- 10454979 TI - Determinants of career structure and advancement among Italian cardiologists. An example of segregation and discrimination against women? SCIC Group. Studio Condizione Italiana Cardiologi. AB - AIMS: The aim of this study was to analyse the processes through which job, career and research-related choices are determined in Italian cardiology, focusing on characteristics such as productivity, gender and family. METHODS AND RESULTS: In June 1996, a questionnaire surveying individual and career-related data was mailed to all members (8000) of the Italian societies of cardiology. Returned questionnaires numbered 1715 (21.4% of the total mailed), 83% were completed by men and 17% by women. For both hospital and academic careers, advancement in rank was influenced by variables denoting productivity, family and individual characteristics. However, men and women showed slightly different patterns. CONCLUSIONS: Promotion to the upper ranks of the hierarchy was highly dependent upon time (once the effects of the covariates were eliminated). This situation is typical of the internal labour market, that is, in institutions in which staff members are ranked on a hierarchical scale according to formal criteria that are 'rigid' and institutionalized, partially sheltered from competition. Therefore, once a member has gained access to the bottom of the hierarchy, the professional career is 'pre-determined' and seniority has an appreciable influence on promotion decisions; in this context, women appear to be at a disadvantage. PMID- 10454981 TI - Use of antisense to study gene expression. Introduction. PMID- 10454982 TI - Achieving antisense inhibition by oligodeoxynucleotides containing N(7)-modified 2'-deoxyguanosine using tumor necrosis factor receptor type 1. AB - Antisense oligodeoxynucleotides (ODNs) are being explored as therapeutic agents for the treatment of many disorders including viral infections, cancers, and inflammatory disorders. In addition, antisense technology can be of great benefit to those attempting to assign function to the multitude of new genes being uncovered in the genomics initiative. However, the demonstration that the gene regulating effects produced by antisense-designed ODNs are attributable to an antisense mechanism of action requires carefully designed experimentation. Critical to the assignment of an antisense mechanism of action is the availability of nuclease-stable ODNs, inside cells, that have a high binding affinity with the target mRNA and modulate gene functions in a sequence-dependent manner. To help us achieve a goal of sequence-specific antisense activity we designed antisense ODNs containing C(5)-propyne-modified 2'-deoxyuracil and N(7) propyne-modified 7-deaza-2'-deoxyguanosine bases and partially modified (phosphorothioate) internucleoside linkages. These modified ODNs were found to have enhanced binding affinity to their target mRNA sequences as well as reduced sequence-independent side effects. We used these ODNs to specifically inhibit p55 tumor necrosis factor receptor type 1 expression and tumor necrosis factor alpha mediated functions in culture assays. PMID- 10454983 TI - Controlling gene expression with 2-5A antisense. AB - Recent work has demonstrated that the activity of a ubiquitous cellular enzyme, ribonuclease L (RNase L), can be harnessed to cleave targeted RNA species. Activation of RNase L is dependent on the presence of 2',5'-linked oligoadenylates (2-5A), usually produced by cells infected with viruses. By conjugating synthetic 2-5A to specific antisense compounds, it is now possible to selectively degrade RNAs in an RNase L-dependent manner, thereby providing an alternative to RNase H-dependent approaches. In this summary, we provide an updated description of the synthesis procedure for constructing these chimeric 2 5A antisense molecules. Examples of successful applications of the 2-5A antisense strategy are described, along with some of the procedures involved in those studies. Several methods are also provided for optimizing compound uptake and analyzing their effects on cells. Finally, we discuss the current body of evidence that supports the contention that RNase L is indeed the primary mediator of 2-5A antisense effects and the possible implications that this has on the future of this therapeutic approach. PMID- 10454984 TI - Use of triplex-forming oligonucleotides and adenoviral constructs for studying the regulation of gene expression. AB - Short synthetic homopyrimidine- or homopurine-rich oligonucleotides can form sequence-specific triplexes with corresponding homopurine-homopyrimidine sites on duplex DNA and block transcription of a target gene in vitro. Such triplex forming oligonucleotides (TFOs) can be rationally designed to target homopurine/homopyrimidine sequences that are often found in eukaryotic genes and thus used to modulate the expression of these genes. The antigene strategy using TFOs has been successfully applied to a number of genes in vitro. In this article we describe methods used in applying this antigene approach to the rat aquaporin 5 (rAQP5) gene. We specifically focus on the selection of TFOs based on the sequence of the target gene and on a novel method employing adenoviruses for delivery of TFOs to cells in vitro. PMID- 10454985 TI - Generation and application of asymmetric hammerhead ribozymes. AB - Most researchers who intend to suppress a particular gene are interested primarily in the application of ribozyme technology rather than its mechanistic details. This article provides some background information and describes a straightforward strategy to generate and test a special design of a ribozyme: the asymmetric hammerhead ribozyme. This version of a hammerhead ribozyme carries at its 5' end the catalytic domain and at its 3' end a relatively long antisense flank that is complementary to the target RNA. Asymmetric hammerhead ribozymes can be constructed via polymerase chain reaction amplification, and rules are provided on how to select the DNA oligonucleotides required for this reaction. In addition to details on construction, we describe how to test asymmetric hammerhead ribozymes for association with the target RNA in vitro, so that RNA constructs can be selected and optimized for fast hybridization with their target RNA. This test can allow one to minimize association problems caused by the secondary structure of the target RNA. Additionally, we describe the in vitro cleavage assay and the determination of the cleavage rate constant. Testing for efficient cleavage is also a prerequisite for reliable and successful application of the technology. A carefully selected RNA will be more promising when eventually used for target suppression in living cells. PMID- 10454986 TI - In situ reverse transcription for detection of hybridization between oligonucleotides and their intracellular targets. AB - It is often important to know that a phenotypic change caused by antisense treatment has occurred because the antisense molecule has specifically hybridized to its intracellular target, rather than by some nonspecific, indirect route. We describe here a method that can be used to detect hybridization of an antisense oligodeoxynucleotide to its intracellular target RNA and, furthermore, to identify the sites at which hybrids are located in situ. Oligodeoxynucleotides are first taken up by the live cell and then cells are fixed and subjected to an in situ reverse transcription reaction. The reverse transcription assay exploits the fact that only oligonucleotides that are hybridized to RNA will act as primers for reverse transcriptase and allow incorporation of labeled nucleotide into cDNA; unhybridized oligonucleotides will not prime reverse transcription. We illustrate this approach by comparing the levels of oligo(dT) hybridized to poly(A) RNA in cells that have taken up the oligo(dT) with and without cationic lipid in the medium. PMID- 10454987 TI - Antisense oligonucleotide delivery with polyhexylcyanoacrylate nanoparticles as carriers. AB - Polyalkylcyanoacrylate nanoparticles are effective colloidal drug carriers and were prepared by an emulsion polymerization process. Antisense oligonucleotides were loaded on the particles by adsorption. A cationic polymer, DEAE-dextran, was incorporated into the particle matrix or a cationic hydrophobic detergent (CTAB) was used to form a lipophilic oligonucleotide ion pair. Enzymatic digestion of the oligonucleotides was almost quantitatively inhibited by this nanoparticle complex and cellular uptake by different cell lines was significantly enhanced. In vivo the biodistribution of the oligonucleotide nanoparticle complex resulted in targeting of oligonucleotides to the liver. Improvements in antisense treatments with nanoparticles were demonstrated for tumor therapy as well as for antiviral applications. PMID- 10454988 TI - Inhibition of oncogene expression using vector-generated RNA antisense. AB - Antisense RNA expression vectors have been developed relatively recently as a means to study the role of specific oncogenes in malignant transformation. In this paper, strategies for the construction of antisense plasmid vectors from commercially available reagents are described. Techniques for the introduction of these vectors into cell lines and tumors are also described and preferred methods for the evaluation of biological effects are presented. Lastly, using specific examples, the limitations and potential artifacts associated with antisense vector use in the study of tumorigenesis are discussed. PMID- 10454989 TI - Antisense transgenics in animals. AB - Antisense transgenesis provides a methodology for ablating gene expression in targeted tissues through the use of tissue-specific or controllable promoters. The two major features to be considered in the design of a construct for injection are (1) the target sequence and (2) the promoter to be used. Information is provided to help the investigator make decisions in these regards. The standard methodology of making transgenics is not replicated but some hints as how best to use a transgenic facility are provided. An overview of methods for studying transgene expression and target gene suppression is given. In summary, antisense transgenesis may provide a more readily achievable method for tissue specific ablation of a gene's function than controllable knockouts provide. PMID- 10454990 TI - Gene identification by shotgun antisense. AB - Shotgun antisense is a technique to make a random set of mutant cells or organisms in such a way that one can select an interesting mutant and then sequence part of the mutated gene within a day. In addition to the fantastic rapidity with which one can identify the mutated gene, there are more advantages of this technique over other mutagenesis techniques: (1) one can identify genes that when completely repressed are lethal; (2) one can select which sets of genes will be mutated; and (3) genes that are expressed from multiple copies can be repressed and thus identified. PMID- 10454992 TI - Color plates PMID- 10454991 TI - Utilization of antisense oligodeoxynucleotides with embryonic tissues in culture. AB - Experimental embryology has long used manipulation of interacting tissues to examine questions of tissue interaction and differentiation. The potential for specific manipulation of gene expression in such tissues has made the utilization of antisense techniques desirable. However, problems with this methodology have discouraged many investigators from using this approach. Selection of target sequences for antisense oligonucleotides, delivery of oligonucleotides into cells or tissues, and the type of modification of the oligonucleotide to be used all present concerns that must be addressed. This paper describes our approach to selection of target sequence and methods of delivery and describes the synthesis of a methoxyethylamidate-modified antisense oligonucleotide that has proved useful in our studies. This approach has enabled us to explore aspects of tissue interaction in the embryonic heart that would have been difficult to explore in a genetic model. PMID- 10454993 TI - The power of genetic analysis, coupled with rigorous biochemistry, offers unparalleled opportunities for further elucidation of both DNA replication and repair. PMID- 10454994 TI - Minichromosome maintenance as a genetic assay for defects in DNA replication. AB - Minichromosome maintenance (mcm) is an effective genetic assay for mutants defective in DNA replication. Two classes of mcm mutants have been identified using this screen: those that differentially affect the activities of certain autonomously replicating sequences (ARSs) and those that uniformly affect the activities of all ARSs. The ARS-specific MCM genes are essential for the initiation of DNA replication. Among these are members of the MCM2-7 family that encode subunits of the preinitiation complex and MCM10, whose gene product interacts with members of the Mcm2-7 proteins. Among the ARS-nonspecific MCM gene products are chromosome transmission factors. Refinement of this genetic assay as a screening tool and further analysis of existing mcm mutants may reveal new replication initiation proteins. PMID- 10454995 TI - Schizosaccharomyces pombe replication and repair proteins: proliferating cell nuclear antigen (PCNA). AB - Schizosaccharomyces pombe has a cell cycle progression with distinctive phases that serves as a perfect model system for investigating DNA replication and repair of eukaryotic cells. Here, we use proliferating cell nuclear antigen (PCNA) of S. pombe to demonstrate how the function of this protein in both DNA replication and repair can be assessed by genetic and biochemical approaches. We describe a method of introducing site-specific mutations into the fission yeast PCNA gene pcn1(+). The in vivo effects of these pcn1 mutants in a strain with a null pcn1 background are described and their in vitro biochemical properties are characterized. Mutants described here are those that are defective in enhancing processivity of DNA polymerase delta, show temperature-sensitive growth, and have increased sensitivity to hydroxyurea (HU), UV and gamma irradiation, and methyl methanesulfonate (MMS). Three mutants that show reduced growth rate in vivo and decreased capacity to enhance polymerase delta DNA synthetic activity and processivity in vitro-pcn1-1, pcn1-5, and pcn1-26-are described as examples of using a genetic approach to identify the biochemical function of replication proteins. One cold-sensitive growth allele, pcn1-3, that has a recessive cold sensitive cdc phenotype and shows sensitivity to HU and UV and gamma irradiation is used as an example of using the genetic approach to reveal the function of replication proteins in repair. The power of combining both biochemical and genetic disciplines is emphasized. Methods for site-directed mutagenesis, in vitro analysis of mutant proteins, and in vivo characterization of mutants in response to UV or gamma irradiation, MMS, HU, and temperature, as well as genetic epistasis are described. Locations of functionally significant residues on the PCNA tertiary structure are summarized. PMID- 10454996 TI - Overexpression of multisubunit replication factors in yeast. AB - Facile genetic and biochemical manipulation coupled with rapid cell growth and low cost of growth media has established the yeast Saccharomyces cerevisiae as a versatile workhorse. This article describes the use of yeast expression systems for the overproduction of complex multipolypeptide replication factors. The regulated overexpression of these factors in yeast provides for a readily accessible and inexpensive source of these factors in large quantities. The methodology is illustrated with the five-subunit replication factor C. Whole-cell extracts are prepared by blending yeast cells with glass beads or frozen yeast with dry ice. Procedures are described that maximize the yield of these factors while minimizing proteolytic degradation. PMID- 10454997 TI - Genetic methods for characterizing the cis-acting components of yeast DNA replication origins. AB - Small circular plasmids containing replication origins and, in some cases, centromeres, can replicate autonomously in the nuclei of all tested yeast species. Because this autonomous replication is dependent on the replication origin within the plasmid, measurements of the efficiency of autonomous replication (by the methods summarized here) permit evaluation of the effects of mutations on origin function. Although alternative methods are available for genetic characterization of replication origins in other organisms, the simplicity of the autonomous replication assay in yeasts has permitted development of the deepest understanding to date of eukaryotic replication origin structure. This information has come primarily from studies with Saccharomyces cerevisiae. However, there are many other yeast species, each with its own variety of replication origins. Use of the methods summarized here to characterize origins in other yeast species is likely to provide additional insights into eukaryotic replication origin structure. PMID- 10454998 TI - In vitro DNA replication in yeast nuclear extracts. AB - Two assays have been developed for studying DNA replication in vitro based on nuclear extracts isolated from budding yeast cells synchronized in S phase. In the first, the template DNA for replication is provided in the form of intact yeast nuclei, usually from cells arrested in G(1). In the second assay, bacterially produced supercoiled plasmid is replicated in an S-phase nuclear extract supplemented with nucleotides and an energy-regenerating system. Semiconservative DNA replication is monitored by substitution of newly synthesized DNA with bromodeoxyuridine 5'-triphosphate (BrdUTP) and density gradient analysis. In addition, neutral-neutral two-dimensional gel analyses and, in the case of nuclei, detection of newly synthesized DNA in replication foci by DIG-dUTP incorporation can be used to monitor replication. PMID- 10454999 TI - DNA repair defects and other (mus)takes in Drosophila melanogaster. AB - Preservation of the structural integrity of DNA in any organism is crucial to its health and survival. Such preservation is achieved by an extraordinary cellular arsenal of damage surveillance and repair functions, many of which are now being defined at the gene and protein levels. Mutants hypersensitive to the killing effects of DNA-damaging agents have been instrumental in helping to identify DNA repair-related genes and to elucidate repair mechanisms. In Drosophila melanogaster, such strains are generally referred to as mutagen-sensitive (mus) mutants and currently define more than 30 genetic loci. Whereas most mus mutants have been recovered on the basis of hypersensitivity to the monofunctional alkylating agent methyl methanesulfonate, they nevertheless constitute a phenotypically diverse group, with many mutants having effects beyond mutagen sensitivity. These phenotypes include meiotic dysfunctions, somatic chromosome instabilities, chromatin abnormalities, and cell proliferation defects. Within the last few years numerous mus and other DNA repair-related genes of Drosophila have been molecularly cloned, providing new insights into the functions of these genes. This article outlines strategies for isolating mus mutations and reviews recent advances in the Drosophila DNA repair field, emphasizing mutant analysis and gene cloning. PMID- 10455001 TI - Chorion gene amplification in Drosophila: A model for metazoan origins of DNA replication and S-phase control. AB - The mechanisms controlling duplication of the metazoan genome are only beginning to be understood. It is still unclear what organization of DNA sequences constitutes a chromosomal origin of DNA replication, and the regulation of origin activity during the cell cycle has not been fully revealed. We review recent results that indicate that chorion gene amplification in follicle cells of the Drosophila ovary is a model for investigating metazoan replication. Evaluation of cis sequence organization and function suggests that chorion loci share attributes with other replicons and provides insights into metazoan origin structure. Moreover, recent results indicate that chorion origins respond to S phase control, but escape mechanisms that inhibit other origins from firing more than once in a cell cycle. Several identified genes that mediate amplification are critical for the cell cycle control of replication initiation. It is likely that further genetic screens for mutations that disrupt amplification will identify the cadre of proteins associated with origins and the regulatory pathways that control their activity. Furthermore, the recent development of methods to detect amplification in situ has uncovered new aspects of its developmental control. Examining this control will reveal links between developmental pathways and the cell cycle machinery. Visualization of amplifying chorion genes with high resolution also represents an opportunity to evaluate the influence of nuclear and chromosome structure on origin activity. The study of chorion amplification in Drosophila, therefore, provides great potential for the genetic and molecular dissection of metazoan replication. PMID- 10455000 TI - Drosophila replication and repair proteins: proliferating cell nuclear antigen (PCNA). AB - Proliferating cell nuclear antigen (PCNA), a protein intimately involved in both replication and repair, has been identified in eukaryotes at all levels of evolution. Is primary sequence, Drosophila melanogaster PCNA is 73% identical to mammalian PCNA. Moreover, it is able to substitute for mammalian PCNA in at least one intricate cell-free replication assay. Mutations in the gene for Drosophila PCNA, including some that are temperature sensitive, have been reported. Procedures are described for the biochemical purification of wild-type PCNA from a population of 6- to 18-h-old Drosophila embryos. Procedures were also developed for purification of unmodified wild-type Drosophila PCNA after induction of expression in Escherichia coli. An NH(2)-terminally His-tagged but otherwise wild type Drosophila PCNA, as well as mutant His-tagged PCNA, were also engineered and purified to apparent homogeneity. Finally, an in situ polyacrylamide gel technique allows DNA polymerase assays to be performed on portions of single adults as well as single Drosophila embryos. This assay should tremendously facilitate systematic genetic studies of metazoan replication and repair. PMID- 10455002 TI - Physical and genetic mapping of mammalian replication origins. AB - The neutral/neutral and neutral/alkaline two-dimensional gel electrophoretic techniques are sensitive physical mapping methods that have been used successfully to identify replication initiation sites in genomes of widely varying complexity. We present detailed methodology for the preparation of replication intermediates from mammalian cells and their analysis by both neutral/neutral and neutral/alkaline two-dimensional gel approaches. The methods described allow characterization of the replication pattern of single-copy loci, even in mammalian cells. When applied to metazoans, initiation is found to occur at multiple sites scattered throughout zones that can be as long as 50 kb, with some subregions being preferred. Although these observations do not rule out the possibility of genetically defined replicators, they offer the alternative or additional possibility that chromosomal context may play an important role in defining replication initiation sites in complex genomes. We discuss novel recombination strategies that can be used to test for the presence of sequence elements critical for origin function if the origin lies in the vicinity of a selectable gene. Application of this strategy to the DHFR locus shows that loss of sequences more than 25 kb from the local initiation zone can markedly affect origin activity in the zone. PMID- 10455003 TI - Histopathology of experimental invasive pulmonary aspergillosis in rats: pathological comparison of pulmonary lesions induced by specific virulent factor deficient mutants. AB - To investigate the pathology of invasive pulmonary aspergillosis (IPA) in detail, a new animal model of IPA was used to compare the histological features induced by three different strains of Aspergillus fumigatus; mutants devoided of exocellular protease and rodlet and their parental strain. To produce an experimental pulmonary lesion of IPA closely mimicking the human disease, suspension of agarose beads containing conidiae of A. fumigatus were used as an inocula to fix infallibly the causative agents in alveoli and rats were treated with a low-dose of immunosuppressive drugs to avoid an induction of agranulocytosis in rodents. There was no significant difference in the mortality of mice with an intravenous injection between these three strains. However, IPA model in the study was successful to demonstrate a significant difference in the histological feature of lungs of infected rats. Pulmonary lesions on the fifth day after infection induced by the rodletless mutant were limited and inflammatory responses were weak when compared to those induced by both no exocellular protease mutant as well as their parental strain. The evidence of rodlet layer in conidia of Aspergilli may play an important role in the physiopathology of the disease in eliminating the neutrophils and macrophages of hosts on the early stage of the infection. PMID- 10455004 TI - Recombinant hyaluronate associated protein as a protective immunogen against Streptococcus equi and Streptococcus zooepidemicus challenge in mice. AB - The capsule of Streptococcus equi, the cause of strangles, and Streptococcus zooepidemicus, associated with equine lower airway disease, plays an important role in evasion of phagocytosis by polymorphonuclear leucocytes. It is composed of hyaluronate, making it non-immunogenic. A hyaluronate associated protein (HAP) from S. equisimilis, whose gene has been sequenced [1], was investigated (a) for its presence in S. equi and S. zooepidemicus and (b) as an immunogen able to interfere with capsule structure and protect against experimental challenge of mice. The purified capsule of S. equi contained a protein of similar molecular mass to the S. equisimilis protein (approximately 53 kDa). Polymerase chain reaction (PCR) using primers derived from the published sequence of S. equisimilis HAP yielded a product from S. equi and S. zooepidemicus of the expected size and susceptibility to restriction endonucleases. Subcloning of two large in frame StuI/SspI fragments of the HAP gene from S. equi, approximately equivalent to the two halves of the molecule, into the expression vector pGEX-3X yielded only the carboxy half in the correct orientation. This latter recombinant produced a GST fusion protein (HAP-GST) of the expected size that was affinity purified. Antibodies in rabbit antiserum to the native protein in purified hyaluronate reacted strongly in immunoblots with HAP-GST. Antiserum to HAP-GST, when soaked into filter paper strips, caused a diminution of capsule production by S. equi cultured on blood agar. Antiserum added into fresh rabbit blood was not opsonic for S. equi. Immunization with HAP-GST significantly reduced rhinitis in Balb/C mice challenged nasally with S. equi and significantly increased survival time and clearance of bacteria in CBA/CA mice challenged intraperitoneally with S. zooepidemicus. PMID- 10455005 TI - Expression of the monocytic differentiation/activation factor P48 in Mycoplasma species. AB - P48 is a 48 kd monocytic differentiation/activation factor previously purified from the conditioned medium of the Reh human pre-B cell leukemia line. It induces differentiation of HL-60 promyelocytic leukemia cells along the monocytic pathway and production of IL1, TNF-alpha and IL6 in human monocytes and monocytic cell lines. Recently our laboratory isolated cDNA clones for P48 from Reh cells and genomic clones from Mycoplasma fermentans DNA and showed that P48 is a M. fermentans gene product. In this paper we report the analysis of P48 expression at the DNA, mRNA and protein levels in different Mycoplasma species. Polymerase Chain Reaction (PCR) analysis of extracted DNA using P48-specific oligonucleotide primers revealed P48 sequences in M. fermentans but not M. hominis, M. iowae, M. genitalium or M. capricolum. Southern analysis of Mycoplasma DNAs revealed hybridizing bands in M. fermentans and M. capricolum under low stringency, but only in M. fermentans under high stringency. Consistent with this, Northern blot studies revealed a single hybridizing transcript in M. fermentans but not in other Mycoplasma species tested. However, Western blot studies with anti-P48 antibodies revealed P48 antigenic material in M. fermentans, as well as M. hominis and M. iowae. These studies demonstrate that the gene for P48 is derived from M. fermentans or a closely related species and is absent in these other species tested. However, the P48 protein exhibits shared antigenic determinants among several Mycoplasma species which presently are of unknown function or significance. P48 is a Mycoplasma -derived immunomodulatory molecule which may be important in Mycoplasma pathophysiology and may be useful in understanding human haematopoietic differentiation and the control of cytokine biosynthesis. PMID- 10455006 TI - Role of Salmonella enterotoxin in overall virulence of the organism. AB - In this study, the Salmonella enterotoxin gene (stn) was mutated by marker exchange mutagenesis, and the overall virulence of the organism was evaluated. Salmonella marker exchange mutants evoked significantly less fluid secretion in mouse intestinal loops compared to that seen with wild-type S. typhimurium. Salmonella mutants were as invasive as wild-type bacteria for HeLa cells; however, their capacity to cause destruction of the intestinal mucosa was impaired, when compared with wild-type bacteria by electron microscopy. Upon oral challenge of mice, the LD(50)of the Salmonella mutants was greater than that for the wild-type bacteria. The fluid secretory potential, as well as a reduction in the LD(50)of these mutants was restored when the mutated stn gene was replaced by the native stn gene sequence. These mutations had no effect on the aerobic growth of these bacteria in minimal or complete medium; anaerobic growth was also not affected. With these studies, we demonstrated that the presence of an intact stn gene contributed significantly to the overall virulence of S. typhimurium in a murine model. PMID- 10455007 TI - The mammalian cell entry operon 1 (mce1) of mycobacterium leprae and mycobacterium tuberculosis. AB - The genome project on Mycobacterium tuberculosis H37Rv has revealed four mammalian cell entry (MTmce1-4) operons putatively involved with entry and survival of mycobacteria in host cells. A homologous operon to the MTmce1 operon was identified in cosmid B983 of Mycobacterium leprae. By comparison with M. tuberculosis, several mutations, or sequencing errors, were predicted at specific sites causing frame shifts in the MLyrbE1A, MLyrbE1B and MLmce1D genes. Using targeted sequencing, sequence errors were identified. The corrected MLmce1 operon sequence appears to be highly homologous to the MTmce1 operon, and similarly encodes eight potential genes. Thus, both M. tuberculosis and M. leprae mce1 operons may be functional and involved in host cell targeting. PMID- 10455008 TI - Use of TUNEL staining to detect apoptotic cells in the lungs of cattle experimentally infected with Pasteurella haemolytica. AB - Lung sections taken from calves with experimental Pasteurella haemolytica respiratory infection exhibited increased numbers of TUNEL positive cells with time after challenge. This finding suggests that P. haemolytica, or toxins and other components released by the organism, induces apoptosis in bovine leukocytes in vivo. By so doing this might impair host defense and contribute in part to the severe pneumonia that characterizes bovine pasteurellosis. PMID- 10455010 TI - Effects of photoreleased cADP-ribose on calcium transients and calcium sparks in myocytes isolated from guinea-pig and rat ventricle. AB - Actions of photoreleased cADP-ribose (cADPR), a novel regulator of calcium induced calcium release (CICR) from ryanodine-sensitive stores, were investigated in cardiac myocytes. Photoreleased cADPR caused an increase in the magnitude of whole-cell calcium transients studied in mammalian cardiac ventricular myocytes (both guinea-pig and rat) using confocal microscopy). Approx. 15 s was required following photorelease of cADPR for the development of its maximal effect. Photoreleased cADPR also increased the frequency of calcium 'sparks', which are thought to be elementary events which make up the whole-cell calcium transient, and were studied in rat myocytes, but had little or no effect on spark characteristics (amplitude, rise time, decay time and distance to half amplitude). The potentiating effects of photoreleased cADPR on both whole-cell transients and the frequency of calcium sparks were prevented by cytosolic application of the antagonist 8-amino-cADPR (5 microM). These experiments, therefore, provide the first evidence in any cell type for an effect of cADPR on calcium sparks, and are the first to show the actions of photoreleased cADPR on whole-cell calcium transients in mammalian cells. The observations are consistent with the effects of cADPR in enhancing the calcium sensitivity of CICR from the sarcoplasmic reticulum in cardiac ventricular myocytes, leading to an increase in the probability of occurrence of calcium sparks and to an increase in whole-cell calcium transients. The slow time-course for development of the full effect on whole-cell calcium transients might be taken to indicate that the influence of cADPR on CICR may involve complex molecular interactions rather than a simple direct action of cADPR on the ryanodine-receptor channels. PMID- 10455011 TI - Glucose enhances insulin promoter activity in MIN6 beta-cells independently of changes in intracellular Ca2+ concentration and insulin secretion. AB - Recent studies have suggested that glucose may activate insulin gene transcription through increases in intracellular Ca(2+) concentration, possibly acting via the release of stored insulin. We have investigated this question by dynamic photon-counting imaging of insulin- and c-fos-promoter-firefly luciferase reporter construct activity. Normalized to constitutive viral promoter activity, insulin promoter activity in MIN6 beta-cells was increased 1.6-fold after incubation at 30 mM compared with 3 mM glucose, but was unaltered at either glucose concentration by the presence of insulin (100 nM) or the Ca(2+) channel inhibitor, verapamil (100 microM). Increases in intracellular [Ca(2+)] achieved by plasma membrane depolarization with KCl failed to enhance either insulin or c fos promoter activity in MIN6 cells, but increased c-fos promoter activity 5-fold in AtT20 cells. Together, these results demonstrate that glucose can exert a direct effect on insulin promoter activity in islet beta-cells, via a signalling pathway which does not require increases in intracellular [Ca(2+)] nor insulin release and insulin receptor activation. PMID- 10455012 TI - Protective role for proteoglycans against cationic lipid cytotoxicity allowing optimal transfection efficiency in vitro. AB - A dependence on proteoglycans for cationic lipid-mediated gene transfer has been suggested in previous studies [Mislick and Baldeschwieler (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 12349-12354; Mounkes, Zhong, Cipres-Palacin, Heath and Debs (1998) J. Biol. Chem. 273, 26164-26170]. We have evaluated the mechanism of proteoglycan involvement in cationic lipid-mediated gene transfer. DNA plasmid uptake and gene expression were studied in wild-type Chinese hamster ovary (CHO) cells (CHO-K1), heparan sulphate-deficient CHO cells (pgsD-677) and proteoglycan deficient CHO cells (pgsB-618). At an optimal ratio of cationic lipid to DNA, a substantial decrease in reporter gene expression was observed in proteoglycan deficient cells compared with that in heparan sulphate-deficient and wild-type cells. However, there were no differences in reporter gene expression between the cell lines when transfected by electroporation. Moreover, all cell lines exhibited equal cationic-lipid-DNA complex uptake activities, as assessed by the measurement of intracellular (32)P-labelled and rhodamine-labelled DNA plasmid. An analysis of reflected-light images of wild-type and proteoglycan-deficient cells suggested that cationic lipids were preferentially toxic to proteoglycan deficient cells. Cell-growth assays confirmed this, showing that cationic lipids exhibited a greater anti-proliferative activity in proteoglycan-deficient cells and in chlorate-treated wild-type cells than in the other cell lines. The growth inhibitory effect of cationic lipids was abrogated by the addition of exogenous sulphated glycosaminoglycans. We conclude that the glycosaminoglycan part of proteoglycans serves a protective role against cationic lipid cytotoxicity, allowing optimal transfection efficiency in vitro. PMID- 10455013 TI - Phosphorylation of Ser-241 is essential for the activity of 3-phosphoinositide dependent protein kinase-1: identification of five sites of phosphorylation in vivo. AB - 3-phosphoinositide-dependent protein kinase-1 (PDK1) expressed in unstimulated 293 cells was phosphorylated at Ser-25, Ser-241, Ser-393, Ser-396 and Ser-410 and the level of phosphorylation of each site was unaffected by stimulation with insulin-like growth factor-1. Mutation of Ser-241 to Ala abolished PDK1 activity, whereas mutation of the other phosphorylation sites individually to Ala did not affect PDK1 activity. Ser-241, unlike the other phosphorylation sites on PDK1, was resistant to dephosphorylation by protein phosphatase 2A(1). Ser-241 lies in the activation loop of the PDK1 kinase domain between subdomains VII and VIII in the equivalent position to the site that PDK1 phosphorylates on its protein kinase substrates. PDK1 expressed in bacteria was active and phosphorylated at Ser-241, suggesting that PDK1 can phosphorylate itself at this site, leading to its own activation. PMID- 10455014 TI - Large-scale production and purification of functional recombinant bovine rhodopsin with the use of the baculovirus expression system. AB - Here we describe a generic procedure for the expression and purification of milligram quantities of functional recombinant eukaryotic integral membrane proteins, exemplified by hexahistidine-tagged bovine rhodopsin. These quantities were obtained with the recombinant baculovirus/Sf9 insect cell-based expression system in large-scale bioreactor cultures with the use of a serum-free and protein-free growth medium. After optimization procedures, expression levels up to 4 mg/l were established. The recombinant rhodopsin could be purified with high overall yield by using immobilized-metal-affinity chromatography on Ni(2+) agarose. After reconstitution into a native lipid environment, the purified protein was functionally indistinguishable from native rhodopsin with regard to the following parameters: spectral absorbance band, structural changes after photoactivation, and G-protein activation. The procedures developed can be adapted to other membrane proteins. The ability to produce and purify tens of milligrams of functional recombinant eukaryotic membrane protein meets the ever increasing demand of material necessary to perform detailed biochemical and structural biophysical studies that are essential in unravelling their working mechanism at a molecular level. PMID- 10455016 TI - Lysine mutagenesis identifies cationic charges of human CYP17 that interact with cytochrome b5 to promote male sex-hormone biosynthesis. AB - Human CYP17 (17alpha-hydroxylase-17,20-lyase; also cytochrome P450c17 or cytochrome P450(17alpha)) catalyses a hydroxylation reaction and another reaction involving the cleavage of a C-C bond (the lyase activity) that is required only for androgen production. Single amino acid mutations in human CYP17, Arg(347)- >His and Arg(358)-->Gln, have been reported to result in the loss of the lyase activity and to cause sexual phenotypic changes in 46XY male patients. By using site-directed mutagenesis we show here that another mutation in human CYP17, Arg(449)-->Ala, for which human variants have yet not been described, also leads to selective lyase deficiency. Furthermore, all the three types of mutants display a loss of responsiveness to cytochrome b(5), an interaction that is essential for lyase activity, and hence male sex-hormone biosynthesis. That the defect could be essentially reversed by lysine mutagenesis has led to the conclusion that the cationic charges on all three residues (at the positions of Arg(347), Arg(358), Arg(449)) are vital for the functional interaction of CYP17 with cytochrome b(5) and that the loss of any one of these cationic charges is catastrophic. PMID- 10455015 TI - Activation of TrkA by nerve growth factor upregulates expression of the cholinergic gene locus but attenuates the response to ciliary neurotrophic growth factor. AB - Nerve growth factor (NGF) stimulates the expression of the cholinergic gene locus, which encodes choline acetyltransferase (ChAT) and vesicular acetylcholine transporter (VAChT), the proteins necessary for the synthesis and storage of the neurotransmitter acetylcholine (ACh). To determine whether this action of NGF is mediated by the p140TrkA NGF receptor (a member of the Trk tyrosine kinase family) we used a murine basal forebrain cholinergic cell line, SN56, stably transfected with rat trkA cDNA. Treatment of these transfectants with NGF activated mitogen-activated protein kinase and increased cytosolic free calcium concentrations, confirming the reconstitution of TrkA-mediated signalling pathways. The expression of ChAT and VAChT mRNA, as well as ACh content, were coordinately up-regulated by NGF in SN56-trkA transfectants. None of these responses occurred in the parental SN56 cells, which do not express endogenous TrkA, indicating that these actions of NGF required TrkA. We previously reported that ciliary neurotrophic factor (CNTF) upregulates the expression of ChAT and VAChT, as well as ACh production, in SN56 cells. The combined treatment of SN56 trkA cells with CNTF and NGF revealed a complex interaction of these factors in the regulation of cholinergic gene locus expression. At low concentrations of CNTF (<1 ng/ml), the upregulation of ACh synthesis evoked by these factors was additive. However, at higher concentrations of CNTF (>1 ng/ml), NGF attenuated the stimulatory effect of CNTF on ChAT and VAChT mRNA and ACh content. This attenuation was not due to interference with early steps of CNTF receptor signalling, as pre-treatment of SN56-trkA cells with NGF did not affect the nuclear translocation of the transcription factor, Stat3, evoked by CNTF. PMID- 10455009 TI - Poly(ADP-ribosyl)ation reactions in the regulation of nuclear functions. AB - Poly(ADP-ribosyl)ation is a post-translational modification of proteins. During this process, molecules of ADP-ribose are added successively on to acceptor proteins to form branched polymers. This modification is transient but very extensive in vivo, as polymer chains can reach more than 200 units on protein acceptors. The existence of the poly(ADP-ribose) polymer was first reported nearly 40 years ago. Since then, the importance of poly(ADP-ribose) synthesis has been established in many cellular processes. However, a clear and unified picture of the physiological role of poly(ADP-ribosyl)ation still remains to be established. The total dependence of poly(ADP-ribose) synthesis on DNA strand breaks strongly suggests that this post-translational modification is involved in the metabolism of nucleic acids. This view is also supported by the identification of direct protein-protein interactions involving poly(ADP-ribose) polymerase (113 kDa PARP), an enzyme catalysing the formation of poly(ADP ribose), and key effectors of DNA repair, replication and transcription reactions. The presence of PARP in these multiprotein complexes, in addition to the actual poly(ADP-ribosyl)ation of some components of these complexes, clearly supports an important role for poly(ADP-ribosyl)ation reactions in DNA transactions. Accordingly, inhibition of poly(ADP-ribose) synthesis by any of several approaches and the analysis of PARP-deficient cells has revealed that the absence of poly(ADP-ribosyl)ation strongly affects DNA metabolism, most notably DNA repair. The recent identification of new poly(ADP-ribosyl)ating enzymes with distinct (non-standard) structures in eukaryotes and archaea has revealed a novel level of complexity in the regulation of poly(ADP-ribose) metabolism. PMID- 10455017 TI - Structural requirement of the calcium-channel subunit alpha2delta for gabapentin binding. AB - Gabapentin [Neurontin, 1-(aminomethyl)cyclohexaneacetic acid] is a novel anticonvulsant drug with a high binding affinity for the Ca(2+)-channel subunit alpha(2)delta. In this study, the gabapentin-binding properties of wild-type and mutated porcine brain alpha(2)delta proteins were investigated. Removal of the disulphide bonds between the alpha(2) and the delta subunits did not result in a significant loss of gabapentin binding, suggesting that the disulphide linkage between the two subunits is not required for binding. Singly expressed alpha(2) protein remained membrane associated. However, alpha(2) alone was unable to bind gabapentin, unless the cells were concurrently transfected with the expression vector for delta, suggesting that both alpha(2) and delta are required for gabapentin binding. Using internal deletion mutagenesis, we mapped two regions [amino acid residues 339-365 (DeltaF) and 875-905 (DeltaJ)] within the alpha(2) subunit that are not required for gabapentin binding. Further, deletion of three other individual regions [amino acid residues 206-222 (DeltaD), 516-537 (DeltaH) and 583-603 (DeltaI)] within the alpha(2) subunit disrupted gabapentin binding, suggesting the structural importance of these regions. Using alanine to replace four to six amino acid residues in each of these regions abolished gabapentin binding. These results demonstrate that region D, between the N-terminal end and the first putative transmembrane domain of alpha(2), and regions H and I, between the putative splicing acceptor sites (Gln(511) and Ser(601)), may play important roles in maintaining the structural integrity for gabapentin binding. Further single amino acid replacement mutagenesis within these regions identified Arg(217) as critical for gabapentin binding. PMID- 10455019 TI - Inwardly rectifying K+ channel Kir7.1 is highly expressed in thyroid follicular cells, intestinal epithelial cells and choroid plexus epithelial cells: implication for a functional coupling with Na+,K+-ATPase. AB - A novel inwardly rectifying K+ channel, Kir7.1, with unique pore properties, was cloned recently. Working in the field of osmoregulation, we have also identified the same human and rat channel and found that the channel is unique not only in its pore sequence but also in its dense localization in the follicular cells of the thyroid gland. Northern blot analysis revealed that the channel message was abundantly expressed in the thyroid gland and small intestine, and moderately in the kidney, stomach, spinal cord and brain. Immunohistochemistry of the rat thyroid, intestine and choroid plexus demonstrated the expression of the channel protein in the follicular cells and epithelial cells, suggesting a role in the regulation of the ion-transporting functions of these specialized cells. The unique pore properties of Kir7.1 make it a strong candidate for the hypothetical low-conductance K+ channel that is functionally coupled with Na+,K(+)-ATPase by recycling K+. We therefore further examined the co-localization of Kir7.1 and Na+,K(+)-ATPase and found that both are localized in the basolateral membrane of the thyroid follicular cell; in the choroid plexus, which is known to be unique in having Na+,K(+)-ATPase in the apical side of the epithelial cells, Kir7.1 was found in the apical membrane, implying a close functional coupling between the channel and Na+,K(+)-ATPase. PMID- 10455018 TI - In vitro analysis of the glucose-transport system in GLUT4-null skeletal muscle. AB - We have characterized the glucose-transport system in soleus muscle from female GLUT4-null mice to determine whether GLUT1, 3 or 5 account for insulin-stimulated glucose-transport activity. Insulin increased 2-deoxyglucose uptake 2.8- and 2.1 fold in soleus muscle from wild-type and GLUT4-null mice, respectively. Cytochalasin B, an inhibitor of GLUT1- and GLUT4-mediated glucose transport, inhibited insulin-stimulated 2-deoxyglucose uptake by >95% in wild-type and GLUT4 null soleus muscle. Addition of 35 mM fructose to the incubation media was without effect on insulin-stimulated 3-O-methylglucose transport activity in soleus muscle from either genotype, whereas 35 mM glucose inhibited insulin stimulated (20 nM) 3-O-methylglucose transport by 65% in wild-type and 99% in GLUT4-null mice. We utilized the 2-N-4-1-(1-azi-2,2,2-triflu oroethyl)benzoyl-1, 3-bis(D-mannose-4-yloxy)-2-propylamine (ATB-BMPA) exofacial photolabel to determine if increased cell-surface GLUT1 or GLUT4 content accounted for insulin stimulated glucose transport in GLUT4-null muscle. In wild-type soleus muscle, cell-surface GLUT4 content was increased by 2.8-fold under insulin-stimulated conditions and this increase corresponded to the increase in 2-deoxyglucose uptake. No detectable cell-surface GLUT4 was observed in soleus muscle from female GLUT4-null mice under either basal or insulin-stimulated conditions. Basal cell-surface GLUT1 content was similar between wild-type and GLUT4-null mice, with no further increase noted in either genotype with insulin exposure. Neither GLUT3 nor GLUT5 appeared to account for insulin-stimulated glucose-transport activity in wild-type or GLUT4-null muscle. In conclusion, insulin-stimulated glucose-transport activity in female GLUT4-null soleus muscle is mediated by a facilitative transport process that is glucose- and cytochalasin B-inhibitable, but which is not labelled strongly by ATB-BMPA. PMID- 10455020 TI - Multisite dephosphorylation and desensitization of conventional protein kinase C isotypes. AB - The generation of antisera specific for the priming phosphorylation sites on protein kinase Calpha (PKCalpha) has permitted analysis of the dephosphorylation of these sites in relation to the down-regulation of the protein. It was demonstrated that these priming sites are subject to agonist-induced dephosphorylation, consistent with inactivation of the protein. Further, the process is shown to be blocked by a PKC inhibitor, indicating a requirement for PKC catalytic activity. This was corroborated by showing that a constitutively active fragment of PKCalpha is able to stimulate the dephosphorylation of wild type PKCalpha in transfected cells. Consistent with a membrane-traffic event, the process controlled by PKC that leads to dephosphorylation is shown to be temperature-sensitive and to correlate with transient accumulation of PKCalpha on cytoplasmic vesicular structures. It was established that the dephosphorylation of priming sites in PKCalpha is not unique and occurs with other conventional PKC isotypes, demonstrating that this is a general desensitization process for this subclass of kinases. The physiological importance of this desensitization is evidenced by the behaviour of PKCbeta1 in U937 cells, where dephosphorylation of the activation loop site is shown to be a function of cell density. PMID- 10455021 TI - Cell-biological assessment of human glucokinase mutants causing maturity-onset diabetes of the young type 2 (MODY-2) or glucokinase-linked hyperinsulinaemia (GK HI). AB - Mutations in the glucokinase (GK) gene cause type-2 maturity-onset diabetes of the young type 2 (MODY-2) and GK-linked hyperinsulinaemia (GK-HI). Recombinant adenoviruses expressing the human wild-type islet GK or one of four mutant forms of GK, (the MODY-2 mutants E70K, E300K and V203A and the GK-HI mutant V455M) were transduced into glucose-responsive insulin-secreting beta-HC9 cells and tested functionally in order to initiate the first analysis in vivo of recombinant wild type and mutant human islet GK. Kinetic analysis of wild-type human GK showed that the glucose S(0. 5) and Hill coefficient were similar to previously published data in vitro (S(0.5) is the glucose level at the half-maximal rate). E70K had half the glucose affinity of wild-type, but similar enzyme activity. V203A demonstrated decreased catalytic activity and an 8-fold increase in glucose S(0.5) when compared with wild-type human islet GK. E300K had a glucose S(0.5) similar to wild-type but a 10-fold reduction in enzyme activity. E300K mRNA levels were comparable with wild-type GK mRNA levels, but Western-blot analyses demonstrated markedly reduced levels of immunologically detectable protein, consistent with an instability mutation. V455M was just as active as wild-type GK, but with a markedly reduced S(0.5). The effects of the different GK mutants on glucose-stimulated insulin release support the kinetic and expression data. These experiments show the utility of a combined genetic, biochemical and cell biological approach to the quantification of functional and structural changes of human GK that result from MODY-2 and GK-HI mutations. PMID- 10455022 TI - Rab6 is phosphorylated in thrombin-activated platelets by a protein kinase C dependent mechanism: effects on GTP/GDP binding and cellular distribution. AB - In platelets and other secretory cells, protein kinase C (PKC) plays a role in exocytosis stimulated by physiological extracellular signals, although its linkage to the secretory machinery is poorly understood. We investigated whether Rab6, a GTP-binding protein that fractionates with platelet alpha-granules, may be involved in linking these processes. We found that Rab6 contains two PKC consensus phosphorylation sites that are evolutionarily conserved. In platelets metabolically labelled with [(32)P]P(i), Rab6 phosphorylation was induced by phorbol esters or by thrombin. This phosphorylation was blocked by a specific PKC inhibitor (Ro-31-8220), but not by a p38 mitogen-activated protein kinase inhibitor (PD-169316). Physiological stimulation of platelets caused a PKC dependent translocation of Rab6 from platelet particulate fractions, nearly doubling the fraction of Rab6 in the cytosol. A human Rab6 isoform (Rab6C) that is preferentially expressed in human platelet RNA was cloned and its phosphorylation by PKC was characterized. Rab6C incorporated up to 2 mol of [(32)P]P(i) per mol of active protein. Rab6C bound GDP and GTP with K(d) values of 113+/-12 and 119+/-27 nM respectively, and hydrolysed GTP at a rate of 100+/ 15 micromol of GTP/mol of Rab6C per min. PKC phosphorylation of Rab6C increased the affinity for GTP by 3-fold, although it had lesser effects on GDP (1.6-fold). Phosphorylation did not alter the GTPase activity. In summary, thrombin activation of platelets leads to PKC-dependent phosphorylation of Rab6 and a translocation of Rab6 to the cytosol. We suggest that PKC phosphorylation may be an important mechanism through which Rab functional interactions in vesicle trafficking and secretion can be altered in response to an external stimulus. PMID- 10455023 TI - Evidence that platelet and tumour heparanases are similar enzymes. AB - In order to enter tissues, blood-borne metastatic tumour cells and leucocytes need to extravasate through the vascular basal lamina (BL), a process which involves a battery of degradative enzymes. A key degradative enzyme is the endoglycosidase heparanase, which cleaves heparan sulphate (HS), an important structural component of the vascular BL. Previously, tumour-derived heparanase activity (which has been shown to be related to the metastatic potential of murine and human melanoma cell lines) was reported to cleave HS and be inhibited by heparin, as distinct from human platelet heparanase, which cleaved both substrates [Nakajima, Irimura and Nicolson (1988) J. Cell Biochem. 36, 157-167]. We recently reported the purification of human platelet heparanase and showed that the enzyme is a 50-kDa endoglucuronidase [Freeman and Parish (1998) Biochem. J. 330, 1341-1350]. We now report the purification and characterization of heparanase activity from highly metastatic rat 13762 MAT mammary adenocarcinoma and human HCT 116 colonic carcinoma cells and from rat liver using essentially the same procedure that was reported for purification of the human platelet enzyme. The rat 13762 MAT tumour enzyme, which has a native M(r) of 45 kDa when analysed by gel-filtration chromatography and by SDS/PAGE, was observed to be an endoglucuronidase that degraded heparin and HS to fragments of the same sizes as the human platelet enzyme does. N-deglycosylation of both the human platelet and rat 13762 MAT tumour enzymes gave, in each case, a 41-kDa band by SDS/PAGE analysis, demonstrating that the observed difference in M(r) between the platelet and tumour enzymes may have been due largely to differences in the relative amounts of N-glycosylation. Two peptides were isolated following Endoproteinase Lys-C digestion of both the human platelet and rat 13762 MAT tumour heparanases and were shown to be highly similar. Both the rat liver and human colonic carcinoma heparanases also degraded both heparin and HS to fragments of the same sizes as the human platelet enzyme does. Western-blot analysis of an SDS/PAGE gel using antibodies raised against human platelet heparanase demonstrated that human platelet, human tumour and rat tumour heparanases were immunochemically cross reactive. In conclusion, because of the similarities in their sizes, substrate specificities, peptide sequences and immunoreactivities, we propose that heparanase activities present in human platelets, rat liver and in rat and human tumour cells are, in fact, mediated by a similar enzyme. PMID- 10455024 TI - cAMP induces co-translational modification of proteins in IPC-81 cells. AB - An elevated cAMP concentration results in growth arrest and protein synthesis dependent apoptosis in the promyelocytic leukaemia cell line IPC-81. A comparison of two-dimensional gels of extracts from these cells labelled with [(35)S]methionine revealed that five distinct protein spots were induced by cAMP in a protein-synthesis-dependent manner. The spots seemed to result from the acidic shift of a precursor protein. The most abundant spot was phospho-actin. The spots induced by cAMP in intact cells were induced by cAMP-dependent protein kinase (cAPK) during the translation in vitro of mRNA from the leukaemia cells. The effect of cAPK was strictly co-translational, none of the spots being induced when cAPK was added after translation. This suggested that the protein spots arose by co-translational phosphorylation catalysed by cAPK. Two of the protein spots, phospho-actin and a protein with a molecular mass of 30 kDa and an isoelectric point of 4.5, were studied further with respect to expression. They were produced during the whole pre-apoptotic period, had cellular half-lives of several hours and were induced by the same concentrations of cAMP analogue that induced apoptosis. It is suggested that the accumulation of co-translationally modified proteins could be important for long-term cAMP signalling. PMID- 10455025 TI - Ethanol and acetaldehyde elevate intracellular [Ca2+] and stimulate microneme discharge in Toxoplasma gondii. AB - One of the first steps in host-cell invasion by the protozoan parasite Toxoplasma gondii occurs when the parasite attaches by its apical end to the target host cell. The contents of apical secretory organelles called micronemes have recently been implicated in parasite apical attachment to host cells. Micronemes are regulated secretory vesicles that discharge in response to elevated parasite intracellular Ca(2+) levels ([Ca2+]i). In the present study we found that ethanol and related compounds produced a dose-dependent stimulation of microneme secretion. In addition, using fluorescence spectroscopy on tachyzoites loaded with the Ca(2+)-sensitive fluorescent dye fura-2, we demonstrated that ethanol stimulated microneme secretion by elevating parasite [Ca2+](i). Furthermore, sequential addition experiments with ethanol and other Ca(2+)-mobilizing drugs showed that ethanol probably elevated parasite [Ca2+](i) by mobilizing Ca(2+) from a thapsigargin-insensitive compartment of neutral pH. Earlier studies have shown that ethanol also elevates [Ca2+](i) in mammalian cells. Thus, because it is genetically tractable, T. gondii might be a convenient model organism for studying the Ca(2+)-elevating effects of alcohol in higher eukaryotes. PMID- 10455026 TI - Palmitoylation of the recombinant human A1 adenosine receptor: enhanced proteolysis of palmitoylation-deficient mutant receptors. AB - Palmitoylation of the recombinant human A(1) adenosine receptor (A(1)AR) expressed in HEK-293 cells is demonstrated by showing that hexahistidine (His(6))/Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys (FLAG) (H/F) A(1)ARs, purified to homogeneity from cells metabolically labelled with [(3)H]palmitate, incorporate tritium into a 38-42 kDa receptor glycoprotein. The amount of palmitoylation is not affected by incubation of cells with the A(1)AR-selective agonist N(6) cyclopentyladenosine (CPA). A(1)AR palmitoylation is abolished by treatment with neutral hydroxylamine or by mutation of Cys-309 to Ala (C(309)-->A). Based on Western blotting and pulse-chase experiments with [(35)S]methionine, at least 90% of wild-type receptors are palmitoylated and turn over with a t1/2 of 6.4 h. Of the C(309)-->A mutated receptors, 40% appear to turn over like wild-type receptors, with a t1/2 of 7.1 h, and 60% appear to be rapidly cleaved to form a 25 kDa receptor fragment that turns over with a t1/2 of 0.8 h. In HEK-293 cell lines expressing similar numbers of wild-type or C(309)-->A mutant A(1)Rs, there is little difference in the kinetics of CPA-induced receptor internalization (1 h), down-regulation (24 h), inhibition of forskolin-stimulated cAMP accumulation, or activation of co-transfected G-protein-activated inward rectifier K(+)/cardiac inward rectifying K(+) (GIRK1/CIR K(+)) channels. Also unaffected by palmitoylation is guanosine 5'-[gamma-thio]-triphosphate ([S]GTP)-sensitive binding to membranes by the agonist (125)I-labelled aminobenzyladenosine. The results suggest that palmitoylation has little effect on receptor-effector coupling, agonist-induced internalization or down-regulation. We speculate that palmitoylation may divert newly synthesized A(1)ARs from a pathway leading to rapid degradation. PMID- 10455027 TI - Elucidation of carbon sources used for the biosynthesis of fatty acids and sterols in the trypanosomatid Leishmania mexicana. AB - Sterols are necessary for the growth of trypanosomatid protozoans; sterol biosynthesis is a potential target for the use and development of drugs to treat the diseases caused by these organisms. This study has used (14)C-labelled substrates to investigate the carbon sources utilized by promastigotes and amastigotes of Leishmania mexicana for the production of sterol [mainly ergosta 5,7,24(24(1))-trien-3beta-ol] and the fatty acid moieties of the triacylglycerol (TAG) and phospholipid (PL) of the organism. The isoprenoid precursor mevalonic acid (MVA) was incorporated into the sterols, and the sterol precursor squalene, by the promastigotes of L. mexicana. However, acetate (the precursor to MVA in most organisms) was a very poor substrate for sterol production but was readily incorporated into the fatty acids of TAG and PL. Other substrates (glucose, palmitic acid, alanine, serine and isoleucine), which are metabolized to acetyl CoA, were also very poor precursors to sterol but were incorporated into TAG and PL and gave labelling patterns of the lipids similar to those of acetate. In contrast, the amino acid leucine was the only substrate to be incorporated efficiently into the squalene and sterol of L. mexicana promastigotes. Quantitative measurements revealed that at least 70-80% of the sterol synthesized by the promastigotes of L. mexicana is produced from carbon provided by leucine metabolism. Studies with the amastigote form of L. mexicana showed that in this case leucine was again the major sterol precursor, whereas acetate was utilized for fatty acid production. PMID- 10455029 TI - Processing and functional display of the 86 kDa heterodimeric penicillin G acylase on the surface of phage fd. AB - The large heterodimeric penicillin G acylase from Alcaligenes faecalis was displayed on the surface of phage fd. We fused the coding sequence (alpha subunit internal peptide-beta subunit) to the gene of a phage coat protein. A modified g3p signal sequence was used to direct the polypeptide to the periplasm. Here we show that a heterodimeric enzyme can be expressed as a fusion protein that matures to an active biocatalyst connected to the coat protein of phage fd, resulting in a phage to which the beta-subunit is covalently linked and the alpha subunit is non-covalently attached. The enzyme can be displayed either fused to the minor coat protein g3p or fused to the major coat protein g8p. In both cases the penicillin G acylase on the phage has the same Michaelis constant as its freely soluble counterpart, indicating a proper folding and catalytic activity of the displayed enzyme. The display of the heterodimer on phage not only allows its further use in protein engineering but also offers the possibility of applying this technology for the excretion of the enzyme into the extracellular medium, facilitating purification of the protein. With the example of penicillin acylase the upper limit for a protein to become functionally displayed by phage fd has been further explored. Polyvalent display was not observed despite the use of genetic constructs designed for this aim. These results are discussed in relation to the pore size being formed by the g4p multimer. PMID- 10455028 TI - Differential regulation of extracellular signal-regulated protein kinases (ERKs) 1 and 2 by cAMP and dissociation of ERK inhibition from anti-mitogenic effects in rabbit vascular smooth muscle cells. AB - The inhibition of extracellular signal-regulated protein kinases (ERKs) is implicated in the negative regulation of vascular smooth muscle cell (VSMC) mitogenesis by cAMP-elevating agents and transforming growth factor beta(1) (TGF beta(1)). These factors inhibited rabbit aortic VSMC mitogenesis induced by platelet-derived growth factor (PDGF)-BB by preventing the entry of cells into S phase. cAMP-elevating agents partly inhibited the late phase (1-4 h) of activation of ERKs 1 and 2 induced by PDGF-BB without inhibiting the early phase of activity (5-15 min) and had no effect on activity induced by basic fibroblast growth factor (bFGF). In contrast, cAMP elevation caused a marked inhibition of early ERK activation induced by angiotensin II and thrombin. TGF-beta(1) had no inhibitory effect on ERK activation induced by PDGF-BB or bFGF. The inhibition of PDGF-BB-stimulated DNA synthesis by either forskolin/3-isobutyl-1-methylxanthine (IBMX) or TGF-beta(1) was not decreased when the agents were added up to 8 h after growth factor. In contrast, the selective ERK kinase inhibitor PD98059 was a weak inhibitor of DNA synthesis; a combination of PD98059 and forskolin/IBMX had an additive inhibitory effect on DNA synthesis. Forskolin/IBMX inhibited the growth factor-induced expression of c-myc mRNA and cyclin D(1) protein, and enhanced the protein expression of p27(kip1). TGF-beta(1) had no effect on the expression of c-myc or p27(kip1) and weakly attenuated the expression of cyclin D(1). These findings support the conclusion that the suppression of VSMC mitogenesis by cAMP and TGF-beta(1) is independent of ERK inhibition. Anti mitogenic effects of cAMP might be primarily mediated by events in late G(1). PMID- 10455030 TI - Phosphotyrosine protein of molecular mass 30 kDa binds specifically to the positively charged region of the pleckstrin N-terminal pleckstrin homology domain. AB - It has been proposed that phosphoinositides and inositol phosphates serve as general ligands for members of the structurally related pleckstrin homology (PH) domain family. The N-terminal PH domain of pleckstrin (N-PH), in contrast with other PH domains, does not bind to any of these ligands with the high affinity expected for a physiological interaction. To examine whether N-PH might instead mediate protein-protein interaction, a fusion protein with glutathione S transferase (GST) expressing N-PH (GST-N-PH) was used to screen [(35)S]methionine metabolically labelled HL-60 and Bac1. 2F5 cell lysates for potential binding partners. A 30 kDa binding protein was identified in both cell lines. Binding to N-PH demonstrated specificity, because binding was approx. 10-fold higher than when an equimolar amount of pleckstrin C-terminal PH domain (GST-C-PH) was used as probe. The 30 kDa protein could also be metabolically labelled with [(32)P]P(i) and proved to be a tyrosine-phosphorylated protein. Binding to N-PH could be specifically inhibited with phosphotyrosine but not with phosphothreonine; the inhibition was concentration-dependent. Site-directed mutagenesis indicated that a positively charged region previously identified as the phosphoinositide-binding site in N-PH and other PH domains, rather than a putative phosphotyrosine-binding region previously identified in structurally similar phosphotyrosine-binding (PTB) domains, served as the binding site. These results suggest that the positively charged region of N-PH has the potential to interact with a protein ligand that contains phosphotyrosine. PMID- 10455031 TI - Anionic phospholipids decrease the rate of slippage on the Ca(2+)-ATPase of sarcoplasmic reticulum. AB - Accumulation of Ca(2+) by the Ca(2+)-ATPase of skeletal-muscle sarcoplasmic reticulum has been measured in reconstituted, sealed vesicles as a function of lipid composition. Measurements were performed in the presence of carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) to eliminate any effects of H(+) transport; in the presence of FCCP, addition of valinomycin had no effect on the level or rate of accumulation of Ca(2+) showing that, in the presence of FCCP, no electrical potential built up across the membrane. Levels of accumulation were low when the phospholipid was dioleoylphosphatidylcholine (DOPC), even though DOPC supports high ATPase activity. Inclusion of 10 mol% anionic phospholipid [dioleoylphosphatidic acid (DOPA) or dioleoylphosphatidylserine (DOPS)] led to higher levels of accumulation of Ca(2+), 10 mol% being the optimum concentration. Cardiolipin or phosphatidylinositol 4-phosphate were more effective than DOPA or DOPS in increasing accumulation of Ca(2+). Effects of anionic phospholipids were seen in the presence of an ATP-regenerating system to remove ADP, and in the presence of phosphate within the reconstituted vesicles to precipitate calcium phosphate. Rates of passive leak of Ca(2+) from the reconstituted vesicles were slow. The Ca(2+)-accumulation process was simulated assuming either simple passive leak of Ca(2+) from the vesicles or assuming slippage on the ATPase, a process in which the phosphorylated intermediate of the ATPase releases bound Ca(2+) on the cytoplasmic rather than the lumenal side of the membrane. The experimental data fitted to a slippage model, with anionic phospholipids decreasing the rate of slippage. PMID- 10455033 TI - Intracellular cleavage of glycosylphosphatidylinositol by phospholipase D induces activation of protein kinase Calpha. AB - Many proteins are anchored to the cell membrane by glycosylphosphatidylinositol (GPI). One of the functions proposed for the GPI anchor is as a possible mediator in signal transduction through its hydrolysis. GPI-specific phospholipase D (GPI PLD) is a secretory protein that is suggested to be involved in the release of GPI-anchored protein from the membrane. In the present study we examined how GPI PLD is involved in signal transduction. When introduced exogenously and overexpressed in cells, GPI-PLD cleaved the GPI anchors in the early secretory pathway, possibly in the endoplasmic reticulum, resulting in an increased production of diacylglycerol. Experiments in vitro and in vivo showed that the association of protein kinase Calpha (PKCalpha) with membranes was increased markedly by expression of GPI-PLD in cells. Furthermore, sucrose-density-gradient centrifugation and immunofluorescence microscopy demonstrated that PKCalpha was translocated to the endoplasmic reticulum membrane in cells expressing GPI-PLD, in contrast with its association with the plasma membrane in cells treated with PMA. We also confirmed that the phosphorylation of c-Fos as well as PKCalpha itself was greatly enhanced by the expression of GPI-PLD. Taken together, these results suggest that GPI-PLD is involved in intracellular cleavage of the GPI anchor, which is a new potential source of diacylglycerol production to activate PKCalpha. PMID- 10455032 TI - Characterization of a flavocytochrome that is induced during the anaerobic respiration of Fe3+ by Shewanella frigidimarina NCIMB400. AB - A 63.9 kDa periplasmic tetrahaem flavocytochrome c(3), designated Ifc(3), was found to be expressed in Shewanella frigidimarina NCIMB400 grown anaerobically with ferric citrate or ferric pyrophosphate as the sole terminal electron acceptor, but not in anaerobic cultures of the bacterium with other respiratory substrates. Ifc(3) was purified to homogeneity and revealed by biochemical, spectroscopic and primary structure analyses to contain four low-spin bis-His ligated c(3)-haems, with midpoint reduction potentials of -73, -141, -174 and 259 mV. A low-potential flavin was present in the form of non-covalently bound FAD; the protein possessed a unidirectional fumarate reductase activity. Disruption of the chromosomal gene encoding Ifc(3), ifcA, did not lead to a significant change in the rate of Fe(3+) reduction in batch culture. However, during such growth the Ifc(3)-deficient mutant produced both a 35 kDa periplasmic c-type cytochrome and a 45 kDa membrane-associated c-type cytochrome at markedly higher levels than did the parent strain. Nucleotide sequencing data from directly upstream of ifcA indicated the presence of an open reading frame encoding a putative outer-membrane beta-barrel protein of 324 amino acid residues. PMID- 10455034 TI - Analysis of agonist function at fusion proteins between the IP prostanoid receptor and cognate, unnatural and chimaeric G-proteins. AB - Direct measures of G-protein activation based on guanine nucleotide exchange and hydrolysis are frequently impossible to monitor for receptors which interact predominantly with G(s)alpha. An isolated FLAG (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) epitope-tagged human IP prostanoid receptor and fusion proteins generated between this form of the receptor and the alpha subunits of its cognate G-protein G(s), G(i1), a G-protein which it fails to activate in co-expression studies, and a chimaeric G(i1)-G(s)6 (a form of G(i1) in which the C-terminal six amino acids were replaced with the equivalent sequence of G(s)) were stably expressed in HEK293 cells. These were detected by [(3)H]ligand-binding studies and by immunoblotting with both an anti-FLAG antibody and with appropriate antisera to the G-proteins. Each construct displayed similar affinity to bind the agonist iloprost. Iloprost stimulated adenylate cyclase activity in clones expressing both IP prostanoid receptor and the IP prostanoid receptor-G(s)alpha fusion protein, and both constructs were shown to interact with and activate endogenously expressed G(s)alpha. Addition of iloprost to membranes of cells expressing the isolated receptor resulted in a small stimulation of high-affinity GTPase activity. Iloprost produced no stimulation of GTPase activity which could be attributed to the IP prostanoid receptor-G(i1)alpha fusion. However, the fusion proteins containing either G(s)alpha or G(i1)-G(s)6alpha produced substantially greater stimulation of GTPase activity than the isolated IP prostanoid receptor. Treatment of cells expressing the IP prostanoid receptor G(i1)-G(s)6alpha fusion protein with a combination of cholera and pertussis toxins allowed direct measurement of agonist activation of the receptor-linked G protein. Normalization of such results for levels of expression of the IP prostanoid receptor constructs demonstrated a 5-fold higher stimulation of GTPase activity when using the G(s)alpha-containing fusion protein and a 9-fold improvement when using the fusion protein containing G(i1)-G(s)6alpha to detect G protein activation compared with expression of the isolated receptor. PMID- 10455035 TI - Overexpression of the FAD-binding domain of the sulphite reductase flavoprotein component from Escherichia coli and its inhibition by iodonium diphenyl chloride. AB - SiR-FP43, the NADPH- and FAD-binding domain of the Escherichia coli sulphite reductase flavoprotein component (SiR-FP), has been overexpressed and characterized. It folds independently, retaining FAD as a cofactor and the catalytic properties associated with the presence of this cofactor. Iodonium diphenyl chloride (IDP) was shown to be a very efficient inhibitor of SiR-FP43 and SiR-FP60, the monomeric form of SiR-FP, containing both FMN and FAD as cofactors (K(i) = 18.5 +/- 5 microM, maximal inactivation rate = 0.053 +/- 0.005 s(-1)). In both cases, inactivation was shown to result from covalent binding of a phenyl group to FAD exclusively, in marked contrast with previous results obtained with cytochrome P450 reductase (CPR), where FMN and a tryptophan were phenylated, but not FAD. However, our kinetic analyses are in agreement with the inhibition mechanism demonstrated with CPR [Tew (1993) Biochemistry 32, 10209 10215]. Nine different FAD phenylated adducts were isolated and, for the first time, two FAD phenylated adducts were identified directly after extraction from a protein. Taken together, our results have shown that flavoprotein inactivation by IDP is not a reliable indicator for a flavin radical intermediate in catalysis. PMID- 10455036 TI - The type II and X cellulose-binding domains of Pseudomonas xylanase A potentiate catalytic activity against complex substrates by a common mechanism. AB - Xylanase A (Pf Xyn10A), in common with several other Pseudomonas fluorescens subsp. cellulosa polysaccharidases, consists of a Type II cellulose-binding domain (CBD), a catalytic domain (Pf Xyn10A(CD)) and an internal domain that exhibits homology to Type X CBDs. The Type X CBD of Pf Xyn10A, expressed as a discrete entity (CBD(X)) or fused to the catalytic domain (Pf Xyn10A'), bound to amorphous and bacterial microcrystalline cellulose with a K(a) of 2.5 x 10(5) M( 1). CBD(X) exhibited no affinity for soluble forms of cellulose or cello oligosaccharides, suggesting that the domain interacts with multiple cellulose chains in the insoluble forms of the polysaccharide. Pf Xyn10A' was 2-3 times more active against cellulose-hemicellulose complexes than Pf Xyn10A(CD); however, Pf Xyn10A' and Pf Xyn10A(CD) exhibited the same activity against soluble substrates. CBD(X) did not disrupt the structure of plant-cell-wall material or bacterial microcrystalline cellulose, and did not potentiate Pf Xyn10A(CD) when not covalently linked to the enzyme. There was no substantial difference in the affinity of full-length Pf Xyn10A and the enzyme's Type II CBD for cellulose. The activity of Pf Xyn10A against cellulose-hemicellulose complexes was similar to that of Pf Xyn10A', and a derivative of Pf Xyn10A in which the Type II CBD is linked to the Pf Xyn10A(CD) via a serine-rich linker sequence [Bolam, Cireula, McQueen-Mason, Simpson, Williamson, Rixon, Boraston, Hazlewood and Gilbert (1998) Biochem J. 331, 775-781]. These data indicate that CBD(X) is functional in Pf Xyn10A and that no synergy, either in ligand binding or in the potentiation of catalysis, is evident between the Type II and X CBDs of the xylanase. PMID- 10455037 TI - Prevalence and geographic disparities in certain congenital anomalies in Quebec: comparison of estimation methods. AB - The purpose of this study was to estimate the prevalence of congenital anomalies in Quebec from MED-ECHO hospitalization records and from records of stillbirths. The results are first compared with those from the Canadian Congenital Anomalies Surveillance System (CCASS) for Quebec and Canada; then the data are examined by period and region of residence. The study results show that, for the congenital anomalies selected for the study, the prevalence rates measured for Quebec from the MED-ECHO data tend to be lower than the prevalence rates for Canada, whereas the rates estimated by CCASS are higher for Quebec than for Canada. The MED-ECHO data cover practically all Quebec births, compared with only 15% coverage by CCASS, and therefore provide a more accurate picture of congenital anomalies in Quebec. PMID- 10455038 TI - Monograph series on aging-related diseases: XII. Parkinson's disease--recent developments and new directions. AB - Parkinson's disease, a chronic progressive disorder of the central nervous system characterized by tremor, rigidity and bradykinesia, usually affects those over the age of 50. Recent developments in research on Parkinson's disease include investigation of the possible role of diet and a growing interest in genetics and inherited factors. The identification of biological markers and other environmental risk factors will play a significant role in future research of the disease as they will be important in the development of prevention strategies. PMID- 10455039 TI - Development of record linkage of hospital discharge data for the study of neonatal readmission. AB - Computerized record linkage has been used increasingly in epidemiologic studies. We developed a multistage, deterministic matching algorithm using various combinations of key variables. Then, from the records for March 1, 1993, to March 31, 1996, contained in the discharge abstract database of the Canadian Institute for Health Information (CIHI), we examined the relation between length of hospital stay at birth and neonatal readmission. A combined use of province/territory of occurrence, 6-digit postal code of residence, date of birth and sex (step 1) matched 88.5% of 26,629 eligible neonatal readmission records with their birth records. Additional use of institution code and chart number or health card number combined with date of birth and sex (step 2 and step 3) increased the matching rate to 93.0%. Compared with the gold standard, step 1 correctly matched 94.4% of the records. We conclude that this deterministic matching algorithm is a feasible and convenient approach to data linkage for the study of neonatal readmission. The linkage strategy may also be helpful in epidemiologic studies of other short-term events PMID- 10455040 TI - Rate and cost of hospitalizations for asthma in Quebec: An analysis of 1988/89, 1989/90 and 1994/95 data. AB - The objectives of this study were to evaluate recent trends in the frequency and length of stay of hospitalization for asthma in the province of Quebec and to estimate the costs of asthma hospitalizations. Data were extracted for persons hospitalized for 30 days or less with a primary diagnosis of asthma in all Quebec short-stay hospitals during the years 1988/89, 1989/90 and 1994/95. There were 1.76 asthma hospitalizations per 1000 persons in Quebec in 1988/89, down to 1.44 in 1989/90 and up again to 1.75 in 1994/95. There was a small decrease in mean length of stay when the three data years were compared. In all three years, the rate of hospitalization was particularly high among young boys. In 1994/95, more hospitalizations occurred during the fall months. We estimated the total cost for asthma hospitalization that year to be $18 to $21 million. PMID- 10455041 TI - The cost of suicide mortality in New Brunswick, 1996. AB - Suicide is a major public health problem in Canada. Suicide deaths affect society by consuming both human lives and economic resources. The present study estimates the economic impact of suicide deaths that occurred in New Brunswick in 1996, using the human capital approach. For the 94 suicide deaths reported, direct costs for health care services, autopsies, funerals and police investigations were $535,158.32. Indirect costs, which estimate the value of lost productivity due to premature death, had the largest economic value, of $79,353,354.56. The mean total cost estimate per suicide death in 1996 was $849,877.80. Although the most significant impact of a suicide death remains the loss of a human life, these results indicate that the economic cost of this public health tragedy in New Brunswick is also great. To our knowledge, this report provides the first complete cost-of-suicide analysis performed in a Canadian province. PMID- 10455042 TI - Canadian national workshop on measurement of sun-related behaviours [Workshop report]. AB - The idea for the workshop described in this paper emerged from recommendations identified at the 1997 Workshop on Research, Policy and Program Planning on Sun Protective Behaviours. At the 1997 workshop, participants developed a set of recommendations for research initiatives related to sun protection efforts in Canada. One of the primary recommendations was to develop a standard set of definitions and core items to assess sun-related behaviours. In response, the authors of this paper agreed to co-chair the 1998 Canadian National Workshop on Measurement of Sun-Related Behaviours. The purpose of this workshop was to develop consensus on a standard set of measures for program evaluation and for monitoring of sun exposure and protective behaviours in Canada. PMID- 10455043 TI - Normal-mode and free-Air gravity constraints on lateral variations in velocity and density of Earth's mantle AB - With the use of a large collection of free-oscillation data and additional constraints imposed by the free-air gravity anomaly, lateral variations in shear velocity, compressional velocity, and density within the mantle; dynamic topography on the free surface; and topography on the 660-km discontinuity and the core-mantle boundary were determined. The velocity models are consistent with existing models based on travel-time and waveform inversions. In the lowermost mantle, near the core-mantle boundary, denser than average material is found beneath regions of upwellings centered on the Pacific Ocean and Africa that are characterized by slow shear velocities. These anomalies suggest the existence of compositional heterogeneity near the core-mantle boundary. PMID- 10455044 TI - Search for cross-species transmission of porcine endogenous retrovirus in patients treated with living pig tissue. The XEN 111 Study Group. AB - Pig organs may offer a solution to the shortage of human donor organs for transplantation, but concerns remain about possible cross-species transmission of porcine endogenous retrovirus (PERV). Samples were collected from 160 patients who had been treated with various living pig tissues up to 12 years earlier. Reverse transcription-polymerase chain reaction (RT-PCR) and protein immunoblot analyses were performed on serum from all 160 patients. No viremia was detected in any patient. Peripheral blood mononuclear cells from 159 of the patients were analyzed by PCR using PERV-specific primers. No PERV infection was detected in any of the patients from whom sufficient DNA was extracted to allow complete PCR analysis (97 percent of the patients). Persistent microchimerism (presence of donor cells in the recipient) was observed in 23 patients for up to 8.5 years. PMID- 10455045 TI - Superconductivity and the c axis spectral weight of high-T(c) superconductors AB - The temperature dependence of the c axis spectral weight (frequency integral of the interplane conductivity) of high transition temperature (high-T(c)) superconductors is shown to be a probe of thermal and quantal fluctuations of the phase of the superconducting order parameter. The behavior of underdoped cuprates is shown to be a natural consequence of superconducting pairing without long ranged phase coherence. Very underdoped cuprates are found to have strong phase fluctuations, even for temperatures much less than the transition temperature. PMID- 10455046 TI - Decreased rates of alluvial sediment storage in the coon creek basin, wisconsin, 1975-93 AB - The total measured rate of alluvial sediment accretion in the agricultural Coon Creek Basin for the period 1975-93 was only about 6 percent of the rate that occurred in the 1930s, but the distributed changes within the basin were highly variable and systematic. Sediment yield (efflux), however, remained relatively constant despite large stream and valley changes within the basin. These observations demonstrate (i) that sediment sources, sinks, and fluxes vary widely over time and space and (ii) that, although improved soil conservation measures have decreased soil erosion, the downstream effects are complex. PMID- 10455047 TI - Switchable tackiness and wettability of a liquid crystalline polymer AB - The spreading velocity of liquids on the surface of a liquid crystalline polymer can be tremendously affected by a slight temperature change. Indeed, a bulk transition between a highly ordered smectic and an isotropic phase induces a sharp change from a rigid to a soft behavior, with consequent effects on the tack properties of the liquid crystalline polymer and on the dewetting dynamics of a liquid on its surface. PMID- 10455048 TI - Chemical "Double Slits": dynamical interference of photodissociation pathways in water AB - Photodissociation of water at a wavelength of 121.6 nanometers has been investigated by using the H-atom Rydberg tagging technique. A striking even-odd intensity oscillation was observed in the OH(X) product rotational distribution. Model calculations attribute this oscillation to an unusual dynamical interference brought about by two dissociation pathways that pass through dissimilar conical intersections of potential energy surfaces, but result in the same products. The interference pattern and the OH product rotational distribution are sensitive to the positions and energies of the conical intersections, one with the atoms collinear as H-OH and the other as H-HO. An accurate simulation of the observations would provide a detailed test of global H(2)O potential energy surfaces for the three (&Xtilde;/A/&Btilde;) contributing states. The interference observed from the two conical intersection pathways provides a chemical analog of Young's well-known double-slit experiment. PMID- 10455049 TI - Alternative dewetting pathways of thin liquid films AB - An alternative pathway for the initiation of dewetting in thin metastable films of partially miscible liquid mixtures is described. In this pathway, phase separation is followed by a dewetting process at the interface between the two phases. Dewetting proceeds (from the sample edges inward) as holes form. The initially smooth film breaks up into droplets at rates much faster than those allowed by classical rupture mechanisms. Marangoni flow appears to be responsible for the initiation of the flow of the dewetting front, and coupling between the flow in the two phases leads to accelerated hole formation. PMID- 10455050 TI - Salt tolerance conferred by overexpression of a vacuolar Na+/H+ antiport in Arabidopsis. AB - Agricultural productivity is severely affected by soil salinity. One possible mechanism by which plants could survive salt stress is to compartmentalize sodium ions away from the cytosol. Overexpression of a vacuolar Na+/H+ antiport from Arabidopsis thaliana in Arabidopsis plants promotes sustained growth and development in soil watered with up to 200 millimolar sodium chloride. This salinity tolerance was correlated with higher-than-normal levels of AtNHX1 transcripts, protein, and vacuolar Na+/H+ (sodium/proton) antiport activity. These results demonstrate the feasibility of engineering salt tolerance in plants. PMID- 10455051 TI - Genetic variation in susceptibility to endocrine disruption by estrogen in mice. AB - Large (more than 16-fold) differences in susceptibility to disruption of juvenile male reproductive development by 17beta-estradiol (E2) were detected between strains of mice. Effects of strain, E2 dose, and the interaction of strain and E2 dose on testes weight and spermatogenesis were all highly significant (P < 0.0001). Spermatid maturation was eliminated by low doses of E2 in strains such as C57BL/6J and C17/Jls. In contrast, mice of the widely used CD-1 line, which has been selected for large litter size, showed little or no inhibition of spermatid maturation even in response to 16 times as much E2. Product safety bioassays conducted with animals selected for fecundity may greatly underestimate disruption of male reproductive development by estradiol and environmental estrogenic compounds. PMID- 10455052 TI - Rapid infection of oral mucosal-associated lymphoid tissue with simian immunodeficiency virus. AB - The early events during infection with an immunodeficiency virus were followed by application of pathogenic simian immunodeficiency virus atraumatically to the tonsils of macaques. Analyses by virologic assays and in situ hybridization revealed that the infection started locally in the tonsils, a mucosal-associated lymphoid organ, and quickly spread to other lymphoid tissues. At day 3, there were few infected cells, but then the number increased rapidly, reaching a high plateau between days 4 and 7. The infection was not detected in the dendritic cell-rich squamous epithelium to which the virus was applied; instead, it was primarily in CD4+ tonsillar T cells, close to the specialized antigen transporting epithelium of the tonsillar crypts. Transport of the virus and immune-activating stimuli across this epithelium would allow mucosal lymphoid tissue to function in the atraumatic transmission of immunodeficiency viruses. PMID- 10455053 TI - Conservatism of ecological niches in evolutionary time AB - Theory predicts low niche differentiation between species over evolutionary time scales, but little empirical evidence is available. Reciprocal geographic predictions based on ecological niche models of sister taxon pairs of birds, mammals, and butterflies in southern Mexico indicate niche conservatism over several million years of independent evolution (between putative sister taxon pairs) but little conservatism at the level of families. Niche conservatism over such time scales indicates that speciation takes place in geographic, not ecological, dimensions and that ecological differences evolve later. PMID- 10455054 TI - AP-2 recruitment to synaptotagmin stimulated by tyrosine-based endocytic motifs. AB - Clathrin-mediated endocytosis is initiated by the recruitment of the clathrin adaptor protein AP-2 to the plasma membrane where the membrane protein synaptotagmin is thought to act as a docking site. AP-2 also interacts with endocytic motifs present in other cargo proteins. Peptides with a tyrosine-based endocytic motif stimulated binding of AP-2 to synaptotagmin and enhanced AP-2 recruitment to the plasma membrane of neuronal and non-neuronal cells. This suggests a mechanism by which nucleation of clathrin-coated pits is stimulated by the loading of cargo proteins. PMID- 10455055 TI - Identification of a mating type-like locus in the asexual pathogenic yeast Candida albicans. AB - Candida albicans, the most prevalent fungal pathogen in humans, is thought to lack a sexual cycle. A set of C. albicans genes has been identified that corresponds to the master sexual cycle regulators a1, alpha1, and alpha2 of the Saccharomyces cerevisiae mating-type (MAT) locus. The C. albicans genes are arranged in a way that suggests that these genes are part of a mating type-like locus that is similar to the mating-type loci of other fungi. In addition to the transcriptional regulators a1, alpha1, and alpha2, the C. albicans mating type like locus contains several genes not seen in other fungal MAT loci, including those encoding proteins similar to poly(A) polymerases, oxysterol binding proteins, and phosphatidylinositol kinases. PMID- 10455057 TI - The evolutionary process, and the process of evolution PMID- 10455058 TI - A role for reactive oxygen species in endothelial cell anoikis. AB - When adherent cells, such as epithelial or endothelial cells, are detached and continuously maintained in suspension, they undergo a form of programmed cell death termed anoikis. We demonstrate that coincident with endothelial cell detachment, there is a dramatic rise in the intracellular level of reactive oxygen species (ROS). Reattachment to a solid surface rapidly attenuates the level of ROS. The mitochondria appear to be the major source of the detachment induced rise in ROS. The change in the intracellular redox state appears to contribute to endothelial anoikis, because treatment with either the cell permeant antioxidant N-acetylcysteine or the flavin protein inhibitor diphenylene iodonium is demonstrated to reduce oxidant levels and protect against subsequent cell death. Similarly, the endogenous intracellular level of ROS is shown to correlate with the extent of cell death. Finally, we demonstrate that the activities of both caspases and of the c-Jun N-terminal kinases are modulated by the rise in intracellular ROS levels. These results suggest that oxidants serve as signaling molecules and regulators of anoikis. PMID- 10455056 TI - Anti-inflammatory properties of cytochrome P450 epoxygenase-derived eicosanoids. AB - The epoxyeicosatrienoic acids (EETs) are products of cytochrome P450 epoxygenases that have vasodilatory properties similar to that of endothelium-derived hyperpolarizing factor. The cytochrome P450 isoform CYP2J2 was cloned and identified as a potential source of EETs in human endothelial cells. Physiological concentrations of EETs or overexpression of CYP2J2 decreased cytokine-induced endothelial cell adhesion molecule expression, and EETs prevented leukocyte adhesion to the vascular wall by a mechanism involving inhibition of transcription factor NF-kappaB and IkappaB kinase. The inhibitory effects of EETs were independent of their membrane-hyperpolarizing effects, suggesting that these molecules play an important nonvasodilatory role in vascular inflammation. PMID- 10455059 TI - Evidence that phospholipid oxidation products and/or platelet-activating factor play an important role in early atherogenesis : in vitro and In vivo inhibition by WEB 2086. AB - The goal of the present studies was to determine whether phospholipid oxidation products and/or platelet-activating factor (PAF) are mediators of early atherogenesis in vivo. Monocyte-endothelial cell interactions have been shown to play an important role in early atherogenesis. We and others have demonstrated that PAF and phospholipid oxidation products, present in atherosclerotic lesions, including 1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine (POVPC), 1 palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC), and 1-palmitoyl-2 epoxyisoprostane E(2)-sn-glycero-3-phosphocholine (PEIPC), mediate the activation of monocytes and/or endothelial cells in vitro. Previous studies have shown that the action of PAF and PAF-like ether-containing phospholipids was inhibited by WEB 2086. We now demonstrate that pretreatment of human aortic endothelial cells with WEB 2086 (10 micromol/L) and several other PAF antagonists before treatment with POVPC and PEIPC but not PGPC prevented the activation of the endothelial cells to bind monocytes. We present evidence to suggest that this inhibition is not mediated by the PAF receptor. The role of bioactive oxidized phospholipids in fatty streak formation was tested using C57BL/6J LDL R-/- mice fed a chow or Western diet for 5 weeks with or without WEB 2086 mixed with drinking water. Quantitative electrospray ionization mass spectrometry showed similar concentrations of WEB 2086 in the plasma of mice on both diets ( approximately 4 to 5 micromol/L); this concentration was inhibitory in vitro. Administration of WEB 2086 did not affect the lipid composition of mouse plasma. However, fatty streak formation was reduced by 62% in animals fed a Western diet, whereas no change was observed in the small lesions of mice on a chow diet. These studies provide evidence that PAF and/or PAF-like phospholipid oxidation products are important mediators of atherosclerotic lesion development in vivo and that specific receptor antagonists for these molecules may represent a novel therapeutic modality. PMID- 10455060 TI - E1A can provoke G1 exit that is refractory to p21 and independent of activating cdk2. AB - E1A can evoke G1 exit in cardiac myocytes and other cell types by displacing E2F transcription factors from tumor suppressor "pocket" proteins and by a less well characterized p300-dependent pathway. Bypassing pocket proteins (through overexpression of E2F-1) reproduces the effect of inactivating pocket proteins (through E1A binding); however, pocket proteins associate with a number of molecular targets apart from E2F. Hence, pocket protein binding by E1A might engage mechanisms for cell cycle reentry beyond those induced by E2F-1. To test this hypothesis, we used adenoviral gene transfer to express various E2F-1 and E1A proteins in neonatal rat cardiac myocytes that are already refractory to mitogenic serum, in the absence or presence of several complementary cell cycle inhibitors-p16, p21, or dominant-negative cyclin-dependent kinase-2 (Cdk2). Rb binding by E2F-1 was neither necessary nor sufficient for G1 exit, whereas DNA binding was required; thus, exogenous E2F-1 did not merely function by competing for the Rb "pocket." E2F-1-induced G1 exit was blocked by the "universal" Cdk inhibitor p21 but not by p16, a specific inhibitor of Cdk4/6; p21 was permissive for E2F-1 induction of cyclins E and A, but prevented their stimulation of Cdk2 kinase activity. In addition, E2F-1-induced G1 exit was blocked by dominant negative Cdk2. Forced expression of cyclin E induced endogenous Cdk2 activity but not G1 exit. Thus, E2F-1-induced Cdk2 function was necessary, although not sufficient, to trigger DNA synthesis in cardiac muscle cells. In contrast, pocket protein-binding forms of E1A induced G1 exit that was resistant to inhibition by p21, whereas G1 exit via the E1A p300 pathway was sensitive to inhibition by p21. Both E1A pathways-via pocket proteins and via p300-upregulated cyclins E and A and Cdk2 activity, consistent with a role for Cdk2 in G1 exit induced by E1A. However, p21 blocked Cdk2 kinase activity induced by both E1A pathways equally. Thus, E1A can cause G1 exit without an increase in Cdk2 activity, if the pocket protein-binding domain is intact. E1A also overrides p21 in U2OS cells, provided the pocket protein-binding domain is intact; thus, this novel function of E1A is not exclusive to cardiac muscle cells. In summary, E1A binding to pocket proteins has effects beyond those produced by E2F-1 alone and can drive S-phase entry that is resistant to p21 and independent of an increase in Cdk2 function. This suggests the potential involvement of other endogenous Rb-binding proteins or of alternative E1A targets. PMID- 10455061 TI - Impaired long-chain fatty acid utilization by cardiac myocytes isolated from mice lacking the heart-type fatty acid binding protein gene. AB - Heart-type fatty acid binding protein (H-FABP), abundantly expressed in cardiac myocytes, has been postulated to facilitate the cardiac uptake of long-chain fatty acids (LCFAs) and to promote their intracellular trafficking to sites of metabolic conversion. Mice with a disrupted H-FABP gene were recently shown to have elevated plasma LCFA levels, decreased cardiac deposition of a LCFA analogue, and increased cardiac deoxyglucose uptake, which qualitatively establishes a requirement for H-FABP in cardiac LCFA utilization. To study the underlying defect, we developed a method to isolate intact, electrically stimulatable cardiac myocytes from adult mice and then studied substrate utilization under defined conditions in quiescent and in contracting cells from wild-type and H-FABP(-/-) mice. Our results demonstrate that in resting and in contracting myocytes from H-FABP(-/-) mice, both uptake and oxidation of palmitate are markedly reduced (between -45% and -65%), whereas cellular octanoate uptake, and the capacities of heart homogenates for palmitate oxidation and for octanoate oxidation, and the cardiac levels of mRNAs encoding sarcolemmal FA transporters remain unaltered. In contrast, in resting H-FABP(-/-) cardiac myocytes, glucose oxidation is increased (+80%) to a level that would require electrical stimulation in wild-type cells. These findings provide a physiological demonstration of a crucial role of H-FABP in uptake and oxidation of LCFAs in cardiac muscle cells and indicate that in H-FABP(-/-) mice the diminished contribution of LCFAs to cardiac energy production is, at least in part, compensated for by an increase in glucose oxidation. PMID- 10455062 TI - Evidence from a novel human cell clone that adult vascular smooth muscle cells can convert reversibly between noncontractile and contractile phenotypes. AB - Smooth muscle cells (SMCs) perform diverse functions that can be categorized as contractile and synthetic. A traditional model holds that these distinct functions are performed by the same cell, by virtue of its capacity for bidirectional modulation of phenotype. However, this model has been challenged, in part because there is no physiological evidence that an adult synthetic SMC can acquire the ability to contract. We sought evidence for this by cloning adult SMCs from human internal thoracic artery. One clone, HITB5, expressed smooth muscle alpha-actin, smooth myosin heavy chains, heavy caldesmon, and calponin and showed robust calcium transients in response to histamine and angiotensin II, which confirmed intact transmembrane signaling cascades. On serum withdrawal, these cells adopted an elongated and spindle-shaped morphology, random migration slowed, extracellular matrix protein production fell, and cell proliferation and [(3)H]thymidine incorporation fell to near 0. Cell viability was not compromised, however; in fact, apoptosis rate fell significantly. In this state, agonist induced elevation of cytoplasmic calcium was even more pronounced and was accompanied by SMC contraction. Readdition of 10% serum completely returned HITB5 cells to a noncontractile, proliferative phenotype. Contractile protein expression increased after serum withdrawal, although modestly, which suggested that the switch to contractile function involved reorganization or sensitization of existing contractile structures. To our knowledge, the physiological properties of HITB5 SMCs provide the first direct demonstration that cultured human adult SMCs can convert between a synthetic, noncontracting state and a contracting state. HITB5 cells should be valuable for characterizing the basis of this critical transition. PMID- 10455063 TI - 11,12-Epoxyeicosatrienoic acid stimulates endogenous mono-ADP-ribosylation in bovine coronary arterial smooth muscle. AB - The role of endogenous ADP-ribosylation in mediating the activation of the Ca(2+) activated K(+) channels was determined in bovine coronary arteries. Endogenous ADP-ribosylation was examined by incubating coronary arterial homogenates or lysates of cultured coronary arterial smooth muscle cells with [adenylate (32)P]NAD. Four (32)P-labeled proteins were observed at 51, 52, 80, and 124 kDa in the homogenates and lysates. This reaction was enhanced by the addition of 11,12-epoxyeicosatrienoic acid (11,12-EET), a cytochrome P450-derived eicosanoid, and GTP to the incubation. By Western blot analysis, 42- and 70-kDa proteins were recognized by specific antibodies against ADP-ribosyltransferase in the coronary arterial homogenates and smooth muscle cell lysate but not in the lysate of endothelial cells. The 52-kDa acceptor protein of endogenous ADP-ribosylation comigrated with a protein ADP-ribosylated by cholera toxin and was recognized and immunoprecipitated by an anti-G(S)alpha antibody. These results suggest that G(S)alpha is one of several acceptors of the ADP-ribose moiety. As shown by the patch-clamp technique, 11,12-EET stimulated the activation of the K(+) channels in the smooth muscle cells, and this activation was completely blocked by novobiocin, vitamin K(1), 3-aminobenzamide, and m-iodobenzylguanidine, inhibitors of endogenous mono-ADP-ribosyltransferases. We conclude that endogenous mono-ADP ribosyltransferases are present in smooth muscle from bovine coronary arteries. These enzymes transfer ADP-ribose to the cellular proteins such as G(S)alpha and may mediate intracellular signal transduction in coronary vascular smooth muscle. In the coronary circulation, the ADP-ribosylation signaling pathway may play an important role in mediating the activation of the K(+) channels induced by 11,12 EET. PMID- 10455064 TI - Mitochondrial electron transport complex I is a potential source of oxygen free radicals in the failing myocardium. AB - Oxidative stress in the myocardium may play an important role in the pathogenesis of congestive heart failure (HF). However, the cellular sources and mechanisms for the enhanced generation of reactive oxygen species (ROS) in the failing myocardium remain unknown. The amount of thiobarbituric acid reactive substances increased in the canine HF hearts subjected to rapid ventricular pacing for 4 weeks, and immunohistochemical staining of 4-hydroxy-2-nonenal ROS-induced lipid peroxides was detected in cardiac myocytes but not in interstitial cells of HF animals. The generation of superoxide anion was directly assessed in the submitochondrial fractions by use of electron spin resonance spectroscopy with spin trapping agent, 5, 5'-dimethyl-1-pyrroline-N-oxide, in the presence of NADH and succinate as a substrate for NADH-ubiquinone oxidoreductase (complex I) and succinate-ubiquinone oxidoreductase (complex II), respectively. Superoxide production was increased 2.8-fold (P<0.01) in HF, which was due to the functional block of electron transport at complex I. The enzymatic activity of complex I decreased in HF (274+/-13 versus 136+/-9 nmol. min(-1). mg(-1) protein, P<0.01), which may thus have caused the functional uncoupling of the respiratory chain and the deleterious ROS production in HF mitochondria. The present study provided direct evidence for the involvement of ROS in the mitochondrial origin of HF myocytes, which might be responsible for both contractile dysfunction and structural damage to the myocardium. PMID- 10455065 TI - Matrix metalloproteinase inhibition during the development of congestive heart failure : effects on left ventricular dimensions and function. AB - The development of congestive heart failure (CHF) is associated with left ventricle (LV) dilation and myocardial remodeling. The matrix metalloproteinases (MMPs) play a significant role in extracellular remodeling, and recent studies have demonstrated increased MMP expression and activity with CHF. Whether increased MMP activity directly contributes to the LV remodeling with CHF remains unknown. Accordingly, this study examined the effects of chronic MMP inhibition (MMPi) on LV size and function during the progression of CHF. Pigs were assigned to the following groups: (1) CHF, rapid pacing for 3 weeks at 240 bpm (n=12); (2) CHF/MMPi, rapid pacing and concomitant MMPi (PD166793, 20 mg/kg per day [n=10]), and (3) control (n=11). With pacing CHF, LV fractional shortening was reduced (19+/-1 versus 45+/-1%), and end-diastolic dimension increased (5.67+/-0.11 versus 3.55+/-0.05 cm), compared with baseline values (P<0.05). In the CHF/MMPi group, LV endocardial shortening increased (25+/-2%) and the end-diastolic dimension was reduced (4.92+/-0.17 cm) compared with CHF-only values (P<0.05). LV midwall shortening was reduced to a comparable degree in the CHF-only and CHF/MMPi groups. LV peak wall stress increased 3-fold with pacing CHF compared with controls and was significantly reduced in the CHF/MMPi group. LV myocardial stiffness was unchanged with CHF but was increased in the CHF/MMPi group. LV myocyte length was increased with pacing CHF compared with controls (180+/-3 versus 125+/-4 microm, P<0.05) and was reduced in the CHF/MMPi group (169+/-4 microm, P<0.05). Basal-state myocyte shortening velocity was reduced with pacing CHF compared with controls (33+/-2 versus 66+/-1 microm/s, P<0.05) and was unchanged in the CHF/MMPi group (31+/-2 microm/s). Using an ex vivo assay system, myocardial MMP activity was increased with pacing CHF and was reduced with chronic MMPi. In summary, concomitant MMPi with developing CHF limited LV dilation and reduced wall stress. These results suggest that increased myocardial MMP activity contributes to LV myocardial remodeling in developing CHF. PMID- 10455066 TI - Estradiol inhibits leukocyte adhesion and transendothelial migration in rabbits in vivo : possible mechanisms for gender differences in atherosclerosis. AB - The mechanism by which estrogens protect against atherosclerosis is not known. We evaluated in vivo whether there is a gender difference in monocyte adhesion and subendothelial migration in hypercholesterolemic rabbits and whether any gender differences observed are due to estradiol. Monocyte adhesion and subendothelial migration were assessed in a blinded fashion by analyzing a standardized segment of aorta using a scanning electron microscope. We also assessed whether estradiol modulates induction of vascular cell adhesion molecule-1 (VCAM-1) protein using Western blot and flow cytometric analyses. We observed that male rabbits develop more monocyte adhesion and subendothelial migration than do female rabbits during hypercholesterolemia. We also observed that oophorectomized rabbits given physiological estradiol supplementation demonstrate fewer adherent and subendothelial monocytes than do oophorectomized rabbits given placebo. VCAM-1 protein expression was increased in aortae obtained from hypercholesterolemic, oophorectomized animals supplemented with placebo, and this increase was attenuated by estradiol. Finally, in cultured rabbit aortic endothelial cells stimulated with lysophosphatidylcholine, we observed an increase in VCAM-1 protein that was inhibited in a concentration-dependent fashion by estradiol. We have demonstrated in vivo that there is a gender difference in monocyte adhesion to endothelial cells and transendothelial migration after hypercholesterolemia and that this gender difference is due in part to estradiol. Our results also suggest that estradiol inhibits monocyte adhesion by inhibiting expression of VCAM-1. PMID- 10455068 TI - Psychosocial factors in the aetiology of coronary heart disease. PMID- 10455069 TI - Aortic root abscess. PMID- 10455070 TI - Stamps in cardiology: de Musset sign. PMID- 10455071 TI - Dynamic cardiomyoplasty: time to wrap it up? PMID- 10455072 TI - Mechanisms of progression in native coronary artery disease: role of healed plaque disruption. AB - OBJECTIVE: To determine the role of healed plaque disruption in the generation of chronic high grade coronary stenosis. METHODS: Coronary arteries obtained at necropsy were perfuse fixed with formal saline for 24 hours at 100 mg Hg. The percentage lumen diameter stenosis was measured in each 3 mm segment containing a plaque, using the lumen size at the nearest histologically normal segment as the reference point. Each segment was prepared for histological examination and stained with Sirius red and immunohistochemistry for smooth muscle actin. Healed disruption was considered to be present when under polarised light there was a break in the yellow-white dense collagen of the cap filled in by more loosely arranged green collagen. Increased smooth muscle density in the green staining areas was required. Each section was read independently by two observers; any segment with discordant views was considered negative. MATERIAL: 31 men aged 51 69 dying suddenly of ischaemic heart disease. 39 coronary arteries were studied containing 256 separate plaques, after excluding coronary arteries with old total occlusions, an acute culprit thrombotic lesion, diffuse disease without normal arterial segments, and arteries related to old myocardial scars. RESULTS: 16 of 99 plaques causing < 20% diameter stenosis had prior disruption. In the 21-50% stenosis range 16 of 86 plaques showed healed disruption. Stenosis >/= 51% by diameter was present in 71 plaques, 52 of which showed a healed disruption pattern. The difference between stenosis < 50% and stenosis >/= 51% was significant by the chi(2) test (p < 0.001). CONCLUSIONS: Subclinical episodes of plaque disruption followed by healing are a stimulus to plaque growth that occurs suddenly and is a major factor in causing chronic high grade coronary stenosis. This mechanism would explain the phasic rather than linear progression of coronary disease observed in angiograms carried out annually in patients with chronic ischaemic heart disease. PMID- 10455073 TI - Plaque erosion is a major substrate for coronary thrombosis in acute myocardial infarction. AB - OBJECTIVE: To evaluate the prevalence of plaque erosion as a substrate for coronary thrombosis. DESIGN: Pathological study in patients with acute myocardial infarction not treated with thrombolysis or coronary interventional procedures. PATIENTS: 298 consecutive patients (189 men, mean (SD) age 66 (11) years; 109 women, 74 (8) years) dying in hospital between 1984 and 1996 from acute myocardial infarction, diagnosed by ECG changes and rise in cardiac enzymes. MAIN OUTCOME MEASURES: Histopathological determination of plaque erosion as substrate for acute thrombosis; location and histological type of coronary thrombosis; acute and healed myocardial infarcts; ventricular rupture. RESULTS: Acute coronary thrombi were found in 291 hearts (98%); in 74 cases (25%; 40/107 women (37.4%) and 34/184 men (18.5%); p = 0.0004), the plaque substrate for thrombosis was erosion. Healed infarcts were found in 37.5% of men v 22% of women (p = 0.01). Heart rupture was more common in women than in men (22% v 10.5%, p = 0.01). The distribution of infarcts, thrombus location, heart rupture, and healed infarcts was similar in cases of plaque rupture and plaque erosion. CONCLUSIONS: Plaque erosion is an important substrate for coronary thrombosis in patients dying of acute myocardial infarction. Its prevalence is significantly higher in women than in men. PMID- 10455074 TI - Cardiac M(2) muscarinic cholinoceptor activation by human chagasic autoantibodies: association with bradycardia. AB - OBJECTIVE: To assess whether exposure of cardiac muscarinic acetylcholine receptors (mAChR) to activating chagasic antimyocardial immunoglobulins results in bradycardia and other dysautonomic symptoms associated with the regulation of heart rate. METHODS: Trypanosoma cruzi infected patients with bradycardia and other abnormalities in tests of the autonomic nervous system were studied and compared with normal subjects. Antipeptide antibodies in serum were demonstrated by an enzyme linked immunosorbent assay using a synthetic 24-mer-peptide corresponding antigenically to the second extracellular loop of the human heart M(2) mAChR. The functional effect of affinity purified antipeptide IgG from chagasic patients on spontaneous beating frequency and cAMP production of isolated normal rat atria was studied. RESULTS: There was a strong association between the finding of antipeptide antibodies in chagasic patients and the presence of basal bradycardia and an altered Valsalva manoeuvre (basal bradycardia: chi(2) = 37.5, p < 0. 00001; Valsalva manoeuvre: chi(2) = 70.0, p < 0.00001). The antipeptide autoantibodies also showed agonist activity, decreasing the rate of contraction and cAMP production. The effects on rat atria resembled the effects of the authentic agonist and those of the total polyclonal chagasic IgG, being selectively blunted by atropine and AF-DX 116, and neutralised by the synthetic peptide corresponding in amino acid sequence to the second extracellular loop of the human M(2) mAChR. CONCLUSIONS: There is an association between circulating antipeptide autoantibodies in chagasic patients and the presence of bradycardia and other dysautonomic symptoms. Thus these autoantibodies are a marker of autoimmune cardiac autonomic dysfunction. The results support the hypothesis that autoimmune mechanisms play a role in the pathogenesis of chagasic cardioneuromyopathy. PMID- 10455075 TI - Infective endocarditis as a complication of a ruptured aneurysm of the sinus of Valsalva. PMID- 10455076 TI - Different microcirculatory and interstitial matrix patterns in idiopathic dilated cardiomyopathy and Chagas' disease: a three dimensional confocal microscopy study. AB - OBJECTIVE: To analyse the morphological aspects of the extracellular matrix and microcirculation to clarify whether chronic Chagas' cardiopathy (CCC) is an accurate model to study the pathogenesis of idiopathic dilated cardiomyopathy (IDCM). DESIGN: Thick histological myocardial sections were prepared to analyse collagen, and microcirculation was examined during confocal laser and light microscopy. SETTING: The specimens were prepared at the pathology service of the Heart Institute of Sao Paulo, Brazil. PATIENTS: Nine control hearts, eight IDCM hearts, and 10 CCC hearts were studied after necropsy. MAIN OUTCOME MEASURES: The number of collagen struts per 100x field, the area of fibrosis (%), and the diameters of arterioles and capillaries were measured in each heart to establish outcome. RESULTS: A smaller number (mean (SD)) of collagen struts was seen in the hearts in the IDCM group (9.1 (4.1)) than in the control (22.4 (3.2)) (p < 0.05) or CCC (15.7 (7.4)) (p > 0.05) groups. Fibrosis was greater in the CCC hearts (13.8 (10.5)%) than in the IDCM hearts (5.9 (6.6)%) (p > 0.05). Major increases in arteriole (65.4 (9.9) microm) and capillary (9.9 (1.7) microm) diameters were seen in the CCC hearts but not in the IDCM hearts (arteriole diameter 40.3 (7.9) microm; capillary diameter 7.9 (1.3) microm). CONCLUSIONS: Hearts demonstrating CCC and IDCM present different extracellular and microvessel alterations. This suggests that distinct pathogenic mechanisms are responsible for each condition and that CCC is not an effective model to study IDCM. PMID- 10455077 TI - Cystic medial degeneration of the aorta is associated with p53 accumulation, Bax upregulation, apoptotic cell death, and cell proliferation. AB - OBJECTIVE: To address a potential role for p53, Bcl2 associated protein X (Bax), and apoptosis in the processes associated with cell turnover during cystic medial degeneration (CMD) of the aorta. METHODS: Histochemical, immunohistochemical, biochemical, and morphometric methods were used to assess the presence and distribution of p53 immunoreactivity (p53-IR) and Bax immunoreactivity (Bax-IR), as well as the presence of apoptosis and tissue repair processes. RESULTS: Immunohistochemical staining disclosed evidence for p53-IR in all specimens in 26.1 (11.5)% of vascular smooth muscle cells (VSMCs) (controls 0.8 (1.3)%; p < 0. 001). Bax-IR was present in all specimens in 10 (5.4)% of medial cells (controls 0.3 (0.5)%; p < 0.001). Medial VSMCs (alpha-actin positive) with cytoplasmic staining for an apoptosis specific protein (c-jun/ASP) were present in 20/20 specimens (0.7 (0.6)% of VSMCs, controls 0%, p < 0.001), whereas terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL) positive VSMCs were present in 17/20 specimens (1 (1.5)% of VSMCs, controls 0%, p < 0.001). The presence of apoptosis was confirmed by electron microscopy and the demonstration of oligonucleosomal DNA fragments after agarose gel electrophoresis. As shown by double labeling and investigation of serial sections, p53-IR, Bax-IR, c-jun/ASP-IR, and positive TUNEL labeling localised to the same compartments of the aortic media, raising a possible role for p53 and Bax in the triggering of apoptosis of VSMC during CMD. MIB1/Ki-67 positive medial VSMCs (alpha-actin positive) and mesenchymal cells (vimentin positive) were present in all specimens (2.5 (2.8)% of medial cells; controls 0.3 (0.9)%, p < 0.001) mainly in the region around the vasa vasorum, indicating that cell regeneration during CMD may originate mainly from the mesenchyme surrounding the vasa vasorum. CONCLUSION: This study shows that the formal pathogenesis of CMD is characterised by p53 accumulation, Bax upregulation, cell death by apoptosis, and cell regeneration. Nevertheless, the precise stimuli of p53 activation and Bax upregulation as well as the role of p53 and apoptosis in the dissection process itself remain elusive. PMID- 10455078 TI - Instantaneous pressure-flow velocity relations of systemic venous return in patients with univentricular circulation. AB - OBJECTIVE: To assess the pressure and flow velocity relations and respiratory variability of the systemic venous and hepatic venous return in patients with univentricular circulation. PATIENTS: 15 selected patients who had undergone cavopulmonary anastomosis (10) or atriopulmonary anastomosis (5). Mean age at operation was 55.1 months (range 9 to 145). Studies were done at 75.5 (32.6) months (mean (SD)) after the operation. SETTING: Tertiary referral centre. METHODS: Patients were studied using simultaneous recordings of ECG, pressure trace, respirometer trace, and pulsed Doppler echocardiography. Mean systemic venous pressure and pulmonary vascular resistance did not differ significantly between the two patient groups. RESULTS: After total cavopulmonary anastomosis, systemic venous pressure tracings showed a flattened pressure curve without any dependence on cardiac or respiratory cycle. After atriopulmonary anastomosis, right atrial pressure tracings showed a significantly higher "a" wave corresponding to atrial contraction, without any respiratory variability. Pulsed Doppler examination of the superior and inferior caval vein and hepatic vein after total cavopulmonary anastomosis did not show a reverse flow after atrial contraction. The inspiratory to expiratory velocity ratio of antegrade flow revealed a significant dependence of flow on changes in intrathoracic pressure in the intra-atrial tunnel, caval veins, and hepatic vein. During expiration, decrease or cessation of antegrade hepatic venous flow was documented. After an atriopulmonary anastomosis, there was a biphasic antegrade venous flow pattern without significant respiratory variation. CONCLUSIONS: After total cavopulmonary anastomosis, there was marked respiratory dependence of systemic and hepatic venous return, whereas after an atriopulmonary anastomosis venous flow pattern varied according to cardiac cycle and pressure trace. The effects of total cavopulmonary anastomosis on venous return might counteract its other haemodynamic advantages. PMID- 10455079 TI - Transcatheter closure of atrial septal defect and interatrial communications with a new self expanding nitinol double disc device (Amplatzer septal occluder): multicentre UK experience. AB - OBJECTIVE: To review the safety and efficacy of the Amplatzer septal occluder for transcatheter closure of interatrial communications (atrial septal defects (ASD), fenestrated Fontan (FF), patent foramen ovale (PFO)). DESIGN: Prospective study following a common protocol for patient selection and technique of deployment in all participating centres. SETTING: Multicentre study representing total United Kingdom experience. PATIENTS: First 100 consecutive patients in whom an Amplatzer septal occluder was used to close a clinically significant ASD or interatrial communication. INTERVENTIONS: All procedures performed under general anaesthesia with transoesophageal echocardiographic guidance. Interatrial communications were assessed by transoesophageal echocardiography with reference to size, position in the interatrial septum, proximity to surrounding structures, and adequacy of septal rim. Stretched diameter of the interatrial communications was determined by balloon sizing. Device selection was based on and matched to the stretched diameter of the communication. MAIN OUTCOME MEASURES: Success defined as deployment of device in a stable position to occlude the interatrial communication without inducing functional abnormality or anatomical obstruction. Occlusion status determined by transoesophageal echocardiography during procedure and by transthoracic echocardiography on follow up. Clinical status and occlusion rates assessed at 24 hours, one month, and three months. RESULTS: 101 procedures were performed in 100 patients (86 ASD, 7 FF, 7 PFO), age 1.7 to 64.3 years (mean (SD), 13.3 (13.9)), weight 9.2 to 100.0 kg (mean 32.5 (23.5)). Procedure time ranged from 30 to 180 minutes (mean 92.4 (29.0)) and fluoroscopy time from 6.0 to 49.0 minutes (mean 16.1 (8.0)). There were seven failures, all occurring in patients with ASD, and one embolisation requiring surgical removal. Immediate total occlusion rate was 20.4%, rising to 84.9% after 24 hours. Total occlusion rates at the one and three month follow up were 92.5% and 98.9%, respectively. Complications were: transient ST elevation (1), transient atrioventricular block (1), presumed deep vein thrombosis (1), presumed transient ischaemic attack (1). CONCLUSIONS: It appears feasible to close interatrial communications and atrial septal defects up to 26 mm stretched diameter safely with the Amplatzer septal occluder. Short term results confirm an early high occlusion rate with no major complications. Careful selection of cases based on the echocardiographic morphology of the ASD and accurate assessment of their stretched diameter is of utmost importance. Further experience with the larger devices and longer term results are required before a firm conclusion regarding its use can be made. PMID- 10455080 TI - Predicting the effect of D,L-sotalol on ventricular tachycardia inducibility from the RR variability response. AB - AIM: To find a rapid way of identifying non-responders to D, L-sotalol in patients with ventricular tachycardia. METHODS: Programmed ventricular stimulation and RR variability were studied in the control state and 10 days after treatment with 160 to 320 mg of D,L-sotalol in 36 consecutive patients with ventricular tachycardia. RESULTS: In 14 patients (group I) D,L-sotalol suppressed ventricular tachycardia inducibility. In 22 patients (group II) sustained ventricular tachycardia remained inducible during D,L-sotalol treatment. The ventricular tachycardia rate was slowed in eight patients and unchanged or accelerated in 14. At baseline, heart rate variability was similar in both groups. During treatment with D,L-sotalol, variables reflecting parasympathetic activity (pNN50, rMSSD, and high frequency amplitude (HF)) increased in both groups: HF increased from (mean (SD)) 75 (68) to 146 (134) in group I (p < 0.05) and from 60 (49) to 125 (79) in group II (p < 0.05). Other variables were unchanged in group I. In group II, the variables associated with sympathetic activity (coefficient of variance (CV), ratio of low frequency amplitude (LF) to HF) decreased significantly: CV decreased from 13 (4) to 9 (2) (p < 0. 001) and LF/HF from 4.74 (3.02) to 3.00 (2.02) (p < 0.05). CONCLUSIONS: The beta blocking effect of D,L-sotalol produced a significant improvement over control values in indices of parasympathetic tone in all treated patients. However, the heart rate variability indices related to sympathetic activity were decreased only in non responders. This effect of D,L-sotalol on heart rate variability could help detect non-responders to the drug and avoid an electrophysiological study. PMID- 10455081 TI - Inadequate sympathovagal balance in response to orthostatism in patients with unexplained syncope and a positive head up tilt test. AB - AIM: To analyse the immediate response of heart rate variability (HRV) in response to orthostatic stress in unexplained syncope. SUBJECTS: 69 subjects, mean (SD) age 42 (18) years, undergoing 60 degrees head up tilt to evaluate unexplained syncope. METHODS: Based on 256 second ECG samples obtained during supine and upright phases, spectral analyses of low (LF) and high frequency (HF) bands were calculated, as well as the LF/HF power ratio, reflecting the sympathovagal balance. All variables were measured just before tilt during the last five minutes of the supine position, during the first five minutes of head up tilt, and just before the end of passive tilt. RESULTS: Symptoms occurred in 42 subjects (vasovagal syncope in 37; psychogenic syncope in five). Resting haemodynamics and HRV indices were similar in subjects with and without syncope. Immediately after assuming the upright posture, adaptation to orthostatism differed between the two groups in that the LF/HF power ratio decreased by 11% from supine (from 2.7 (1.5) to 2.4 (1.2)) in the positive test group, while it increased by 11.5% (from 2.8 (1.5) to 3.1 (1.7)) in the negative test group (p = 0.02). This was because subjects with a positive test did not have the same increment in LF power with tilting as those with a negative test (11% v 28%, p = 0.04), while HF power did not alter. A decreased LF/HF power ratio persisted throughout head up tilt and was the only variable found to discriminate between subjects with positive and negative test results (p = 0.005, multivariate analysis). During the first five minutes of tilt, a decreased LF/HF power ratio occurred in 33 of 37 subjects in the positive group and three of 27 in the negative group. Thus a decreased LF/HF ratio had 89% sensitivity, 89% specificity, a 92% positive predictive value, and an 86% negative predictive value. CONCLUSIONS: Through the LF/HF power ratio, spectral analysis of HRV was highly correlated with head up tilt results. Subjects developing syncope late during continued head up tilt have a decrease in LF/HF ratio immediately after assuming the upright posture, implying that although symptoms have not developed the vasovagal reaction may already have begun. This emphasises the major role of the autonomic nervous system in the genesis of vasovagal (neurally mediated) syncope. PMID- 10455082 TI - Incidence and modes of onset of early reinitiation of atrial fibrillation after successful internal cardioversion, and its prevention by intravenous sotalol. AB - OBJECTIVES: To study the incidence and mode of onset of early reinitiation of atrial fibrillation (ERAF) following successful internal cardioversion of chronic atrial fibrillation, and to determine the effects of sotalol in the prevention of ERAF. DESIGN: The incidence and modes of onset of ERAF and the acute effects of intravenous sotalol in the prevention of ERAF were studied retrospectively. SETTING: Electrophysiology laboratory at a university teaching hospital. PATIENTS: 64 patients, mean (SD) age 62 (10) years, who underwent internal cardioversion of chronic atrial fibrillation (mean duration of atrial fibrillation 31 (39) months). MAIN OUTCOME MEASURES: ECGs and intracardiac electrograms recorded during the internal cardioversion of atrial fibrillation using 3/3 ms biphasic, R wave synchronised shocks. RESULTS: 52 patients (81%) had successful electrical cardioversion, and 20 (31%) of these had ERAF during the procedure. There was no clinical predictor for the occurrence of ERAF. Fifty eight episodes of ERAF were observed. Five ERAF episodes (9%) had preceding bradycardia and 53 (91%) of these were triggered by atrial premature beats with normal preceding heart rate. Atrial premature beats that reinitiated atrial fibrillation had a shorter coupling interval (333 (43) ms v 396 (100), p < 0.001) and a lower prematurity index (0.44 (0.11) v 0. 55 (0.14), p < 0.001) than those that did not reinitiate atrial fibrillation. Repeated shock delivery and increasing the defibrillation energy did not prevent ERAF. Intravenous sotalol infusion decreased the numbers of atrial premature beats and prolonged their coupling interval, and prevented ERAF after repeated defibrillation in 83% of patients with ERAF. CONCLUSIONS: ERAF is a significant clinical problem after successful internal cardioversion of chronic atrial fibrillation, and was observed in up to 31% of patients. In most episodes, ERAF was triggered by short coupling atrial premature beats with preceding normal heart rate. Intravenous sotalol was effective in preventing ERAF in most cases. PMID- 10455083 TI - Cost effectiveness of HMG-CoA reductase inhibitor (statin) treatment related to the risk of coronary heart disease and cost of drug treatment. AB - OBJECTIVES: To estimate the cost effectiveness of statin treatment in preventing coronary heart disease (CHD) and to examine the effect of the CHD risk level targeted and the cost of statins on the cost effectiveness of treatment. DESIGN: Cohort life table method using data from outcome trials. MAIN OUTCOME MEASURES: The cost per life year gained for lifelong statin treatment at annual CHD event risks of 4.5% (secondary prevention) and 3.0%, 2.0%, and 1.5% (all primary prevention), with the cost of statins varied from pound 100 to pound 800 per year. RESULTS: The costs per life year gained according to annual CHD event risk were: for 4.5%, pound 5100; 3.0%, pound 8200; 2.0%, pound 10 700; and 1.5%, pound 12 500. Reducing the cost of statins increases cost effectiveness, and narrows the difference in cost effectiveness across the range of CHD event risks. CONCLUSIONS: At current prices statin treatment for secondary prevention, and for primary prevention at a CHD event risk 3.0% per year, is as cost effective as many treatments in wide use. Primary prevention at lower CHD event risks (< 3.0% per year) is less cost effective and unlikely to be affordable at current prices and levels of health service funding. As the cost of statins falls, primary prevention at lower risk levels becomes more cost effective. However, the large volume of treatment needed will remain a major problem. PMID- 10455084 TI - Safety of the transpulmonary ultrasound contrast agent NC100100: a clinical and haemodynamic evaluation in patients with suspected or proved coronary artery disease. AB - OBJECTIVE: To evaluate the clinical and haemodynamic safety of NC100100, a new transpulmonary ultrasound contrast agent intended for vascular use. DESIGN: Pulmonary artery pressures and gas exchange, left ventricular and systemic blood pressure and ECG were measured at baseline, after saline injection, and after each of two increasing doses of NC100100 injected intravenously. PATIENTS: 30 patients who were evaluated for suspected coronary artery disease. RESULTS: No change was detected in any of the haemodynamic variables, or in haematological or clinical chemical parameters. Blood gases were unchanged, as were heart rhythm and arterial oxygen saturation. No serious adverse reactions were reported. CONCLUSIONS: NC100100 appeared to be haemodynamically inert and safe in patients with coronary artery disease. PMID- 10455085 TI - Adaptive mechanisms of arterial and venous coronary bypass grafts to an increase in flow demand. AB - OBJECTIVE: To compare the mechanisms by which arterial and venous grafts increase their flow during pacing induced tachycardia, early and later after coronary bypass surgery. DESIGN: 43 grafts (13 epigastric artery, 15 mammary artery, 15 saphenous vein) evaluated early (9 (3) days (mean (SD)) after bypass surgery were compared with 41 other grafts (15 epigastric, 11 mammary, 15 saphenous vein) evaluated later after surgery (mean 23 months, range 6 to 168 months) by quantitative angiography and intravascular Doppler velocity analysis during atrial pacing. Controls were 17 normal coronary arteries. RESULTS: Baseline graft flow tended to be lower later after surgery than early (41 (16) v 45 (21) ml/min, NS). Blood flow increased during pacing by 30 (16)% early after surgery, less than later after surgery (+46 (18)%, p < 0.001) and less than in normal coronary arteries (+54 (27)%, p < 0.001 v early grafts; NS v late grafts). There was no difference between venous and arterial grafts. No significant vasodilatation was observed during pacing early after surgery in arterial and venous grafts. Later after surgery, significant vasodilatation was observed only in arterial grafts (mammary and epigastric grafts), from 2.41 (0.37) to 2.53 (0. 41) mm (+5.1% v basal, p < 0.001). Early after surgery and in venous grafts later after surgery, the increase in flow was entirely due to an increase in velocity. In later arterial grafts, the relative contribution of the increase in velocity to the increase in flow during pacing was lower in arterial grafts (70 (22)%) than in venous grafts (102 (11)%, p < 0.001) and similar to normal coronary arteries (68 (28)%). CONCLUSIONS: Early and later after surgery, arterial grafts and venous grafts both increase their flow similarly during pacing. Early arterial grafts and venous grafts increase their flow only through an increase in velocity. Later after surgery, arterial grafts act as more physiological conduits and increase their flow in the same way as normal coronary arteries, through an increase in velocity and calibre mediated by the endothelium. PMID- 10455086 TI - Sleep apnoea in ischaemic heart disease: differences between acute and chronic coronary syndromes. AB - OBJECTIVE: To evaluate the incidence of sleep apnoea in acute and chronic coronary syndromes. DESIGN: Analysis of sleep and breathing characteristics in a polysomnographic study. SETTING: Cardiology department in tertiary referral centre. PATIENTS: 23 patients were studied soon after acute myocardial infarction (group 1), 22 after clinical stabilisation of unstable angina (group 2), and 22 who had stable angina (group 3). Conditions liable to cause sleep apnoea, such as obesity, chronic obstructive pulmonary disease, neurological disorders, or the use of benzodiazepines, were exclusion criteria. MAIN OUTCOME MEASURES: Sleep apnoea and hypopnoea, oxygen saturation, and sleep indices evaluated soon after clinical stabilisation in groups 1 and 2 and also in group 3. RESULTS: Sleep apnoea, mainly of the central type, was equally present in groups 1 and 2 (mean (SD) apnoea-hypopnoea index: 11.10 (19.42) and 14.79 (20.52), respectively) and more severe than in group 3 (2.82 (6.43), p < 0. 01). Total time spent at SaO(2) < 90%, although significantly greater in group 1 and 2 (0.89 (2.4), 1.42 (3.23) min) than in group 3 (0.01 (0.05) min, p < 0.05), was clinically irrelevant. More arousals per hour of sleep (p < 0.05) were detected in group 1 (5.15 (3.71)) and group 2 (5.31 (2.14)) than in group 3 (2.83 (1.51)). CONCLUSIONS: Sleep apnoea, chiefly of the central type, not only characterises acute myocardial infarction, as found by others, but also unstable angina studied after recent stabilisation. Patient selection by exclusion of other causes of breathing disorders shows that coronary disease related apnoea is absent in the chronic coronary syndrome. In acute syndromes the lack of clinically significant apnoea related oxygen desaturation, together with the low associated incidence of major ischaemic and arrhythmic events, suggests that sleep apnoea is benign in these circumstances, despite a worsening of sleep quality. PMID- 10455088 TI - Covered stent to treat co-existent coarctation and aneurysm of the aorta in a young man. PMID- 10455087 TI - Loss of the normal coupling between the anaerobic threshold and insulin sensitivity in chronic heart failure. AB - OBJECTIVE: To explore whether the anaerobic threshold, a measure of the balance between aerobic and anaerobic cellular metabolism, is related to whole body insulin sensitivity in healthy individuals and in patients with chronic heart failure, which involves is an imbalance of aerobic and anaerobic metabolism. DESIGN: Case-control study. SETTING: A teaching hospital department specialising in heart failure. PATIENTS: 20 healthy individuals (mean (SEM) age 55.2 (2.7) years) and 36 patients with chronic heart failure (59.1 (2.0) years, New York Heart Association class I-IV, anaerobic threshold 11.8 (0. 7) ml/kg/min, left ventricular ejection fraction 26 (2)%). INTERVENTIONS: An intravenous glucose tolerance test for assessment of insulin sensitivity (minimal model analysis) and a maximum treadmill exercise test for assessment of the anaerobic threshold, derived from measurement of oxygen consumption and carbon dioxide output. MAIN OUTCOME MEASURES: Relation between insulin sensitivity and the anaerobic threshold in patients with chronic heart failure. RESULTS: While anaerobic threshold was positively correlated with insulin sensitivity in healthy controls (r = 0.72, p < 0.001), no such relation was observed in patients with chronic heart failure. In stepwise multiple linear regression analyses of variables in healthy individuals, insulin sensitivity emerged as the only predictor of anaerobic threshold (standardised coefficient = 0.72, p < 0.001), while fasting insulin, incremental insulin area, and total body fat (dual photon x ray absorptiometry) failed to enter into final models (joint R = 0.52, p < 0.001). CONCLUSIONS: In healthy individuals, whole body insulin sensitivity is related, or "coupled, " to the anaerobic threshold. The absence of such metabolic coupling in patients with chronic heart failure provides further evidence of disturbed cellular metabolism in patients with this condition. PMID- 10455089 TI - Inappropriately low plasma leptin concentration in the cachexia associated with chronic heart failure. AB - BACKGROUND: Cardiac cachexia is a syndrome of generalised wasting which caries a poor prognosis and is associated with raised plasma concentrations of tumour necrosis factor alpha (TNFalpha). TNFalpha increases secretion of leptin, a hormone which decreases food intake and increases energy expenditure. OBJECTIVE: To determine whether an inappropriate increase in plasma leptin concentration contributes to the cachexia of chronic heart failure. DESIGN: Retrospective case control study. SETTING: Tertiary referral cardiology unit. PATIENTS: 110 human subjects comprising 29 cachectic chronic heart failure patients, 22 non-cachectic chronic heart failure patients, 33 patients with ischaemic heart disease but normal ventricular function, and 26 healthy controls. INTERVENTIONS: Measurement of: body fat content by skinfold thickness (cachectic males < 27%, females < 29%); plasma leptin, TNFalpha, and noradrenaline (norepinephrine); central haemodynamics in chronic heart failure patients at right heart catheterisation. MAIN OUTCOME MEASURES: Plasma leptin concentration corrected for body fat content, plasma TNFalpha and noradrenaline concentration, and central haemodynamics. RESULTS: Mean (SEM) plasma leptin concentrations were: 6.2 (0.6) ng/ml (cachectic heart failure), 16.9 (3.6) ng/ml (non-cachectic heart failure), 16.8 (3.0) ng/ml (ischaemic heart disease), and 18.3 (3.5) ng/ml (control) (p < 0.001 for cachectic heart failure v all other groups). Plasma leptin concentration remained significantly lower in the cachectic heart failure group even after correcting for body fat content and in spite of significantly increased TNFalpha concentrations. Thus plasma leptin was inappropriately low in cachectic chronic heart failure in the face of a recognised stimulus to its secretion. There was no significant correlation between plasma leptin, New York Heart Association class, ejection fraction, or any haemodynamic indices. CONCLUSIONS: Leptin does not contribute to the cachexia of chronic heart failure. One or more leptin suppressing mechanisms may operate in this syndrome-for example, the sympathetic nervous system. PMID- 10455091 TI - Autonomic control of the cerebral circulation during normal and impaired peripheral circulatory control. AB - OBJECTIVE: To determine whether oscillations in the cerebrovascular circulation undergo autonomic modulation in the same way as cardiovascular oscillations. DESIGN: Cardiovascular and cerebrovascular oscillations were monitored at rest and during sympathetic stimulation (head up tilt). The association with and transmission of the oscillations in the sympathetic (low frequency, LF) and respiratory (high frequency, HF) bands was assessed. SUBJECTS: 13 healthy volunteers, 10 subjects with vasovagal syncope, and 12 patients with complicated non-insulin dependent diabetes mellitus. MAIN OUTCOME MEASURES: Power spectrum analysis of cerebral blood flow velocity, arterial blood pressure, and heart rate. Coherence analysis was used to study the association between each pair of oscillations. Phase analysis showed the delay of the oscillations in the cardiovascular signals with respect to the cerebrovascular signals. RESULTS: The power in the sympathetic (LF) components in all the oscillations increased during head up tilt (p < 0.01) in the controls and in the subjects with vasovagal syncope, but not in patients with diabetes. Significant coherence (> 0.5) in the LF band was present between cerebrovascular and cardiovascular oscillations in most of the controls and in subjects with vasovagal syncope, but not in the diabetic patients (< 50% of the patients). In the LF band, cerebrovascular oscillations preceded the cardiovascular oscillations (p < 0.05) at rest in all groups: the phase shifts were reduced (p < 0.05) during head up tilt for all cardiovascular signals in healthy and syncopal subjects, but only for heart rate in diabetic patients. CONCLUSIONS: The cerebrovascular resistance vessels are subject to autonomic modulation; low frequency oscillations in cerebral blood flow velocity precede the resulting fluctuations in other cardiovascular signals. Autonomic neuropathy and microvascular stiffness in diabetic patients reduces this modulation. PMID- 10455090 TI - Estimation of cardiac reserve by peak power: validation and initial application of a simplified index. AB - OBJECTIVES: To validate a simplified estimate of peak power (SPP) against true (invasively measured) peak instantaneous power (TPP), to assess the feasibility of measuring SPP during exercise and to correlate this with functional capacity. DESIGN: Development of a simplified method of measurement and observational study. SETTING: Tertiary referral centre for cardiothoracic disease. SUBJECTS: For validation of SPP with TPP, seven normal dogs and four dogs with dilated cardiomyopathy were studied. To assess feasibility and clinical significance in humans, 40 subjects were studied (26 patients; 14 normal controls). METHODS: In the animal validation study, TPP was derived from ascending aortic pressure and flow probe, and from Doppler measurements of flow. SPP, calculated using the different flow measures, was compared with peak instantaneous power under different loading conditions. For the assessment in humans, SPP was measured at rest and during maximum exercise. Peak aortic flow was measured with transthoracic continuous wave Doppler, and systolic and diastolic blood pressures were derived from brachial sphygmomanometry. The difference between exercise and rest simplified peak power (Delta SPP) was compared with maximum oxygen uptake (VO(2)max), measured from expired gas analysis. RESULTS: SPP estimates using peak flow measures correlated well with true peak instantaneous power (r = 0.89 to 0.97), despite marked changes in systemic pressure and flow induced by manipulation of loading conditions. In the human study, VO(2)max correlated with Delta SPP (r = 0.78) better than Delta ejection fraction (r = 0.18) and Delta rate-pressure product (r = 0.59). CONCLUSIONS: The simple product of mean arterial pressure and peak aortic flow (simplified peak power, SPP) correlates with peak instantaneous power over a range of loading conditions in dogs. In humans, it can be estimated during exercise echocardiography, and correlates with maximum oxygen uptake better than ejection fraction or rate-pressure product. PMID- 10455092 TI - Cardiac rehabilitation: socially deprived patients are less likely to attend but patients ineligible for thrombolysis are less likely to be invited. AB - OBJECTIVE: To identify factors associated with the uptake of cardiac rehabilitation following acute myocardial infarction. DESIGN: Retrospective analysis using multivariate logistic regression modelling. SETTING: Two large teaching hospitals in Nottingham. PATIENTS: Cohorts of patients admitted with acute myocardial infarction in 1992 and 1996. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Factors in multivariate analysis found to be associated with attendance at cardiac rehabilitation. Use of secondary prevention in those who were and were not invited and those who did and did not attend cardiac rehabilitation. RESULTS: 58% of all patients were offered cardiac rehabilitation. Attendance rates were 60% in 1992 and 74% in 1996. Invitations were more likely to be offered to younger patients, those who had received thrombolysis, and to patients admitted to one of the two Nottingham hospitals. Use of secondary prevention was only 48% in 1992 but this increased to 80% in 1996. Patients not receiving secondary prevention were less likely to be invited to cardiac rehabilitation. Social deprivation was the only factor significantly associated with poor uptake of cardiac rehabilitation in both years. There was no difference in the use of secondary prevention between those who did and did not attend cardiac rehabilitation. CONCLUSION: Those invited to attend a cardiac rehabilitation programme are likely to be in a good prognosis group, comprising those who are young and have received thrombolysis. Those at greatest risk, particularly patients from socially deprived areas, seem to be missing out on the potential benefits of cardiac rehabilitation. High risk patients should be specifically targeted to ensure that they are invited to, and encouraged to, attend a programme of cardiac rehabilitation. PMID- 10455093 TI - Technician run open access exercise electrocardiography. AB - OBJECTIVE: To evaluate the safety, efficacy, and feasibility of the technician run open access exercise electrocardiography service at Freeman Hospital. DESIGN: Questionnaire analysis of the responses of the general practitioners of randomly selected patients who used the service. SETTING: A tertiary care cardiac centre, providing an open access service to general practitioners in the community. PATIENTS: 269 patients randomly selected from 552 who underwent open access exercise electrocardiography over a 2.5 year period. OUTCOME MEASURES: Utilisation of service: the reasons for referral, whether the service was optimally used by the general practitioners, and its effect on their management practice; effect on number of cardiology referrals; benefit to the patients; safety, efficacy, and feasibility of a technician run service; general practitioners' assessment of the service. RESULTS: 147 of 178 general practitioners (82.6%) responded to the questionnaire, on 247 of 269 patients (91. 8%). General practitioners used the service for diagnosing ischaemic heart disease in 72.5% of cases, for prognostic purposes in 17.8%, or both in 5.3%. In 197 cases (79.8%), the general practitioners felt that the service had changed the way they managed their patients. The exercise test was positive in 90 patients (36.5%) and identified 38 as at high risk. The service was effective in optimising the cardiology service by reducing referrals by 47%. CONCLUSIONS: The service was used by general practitioners primarily for diagnosing ischaemic heart disease and not so often for prognosis. The utilisation of the service was optimal as assessed by the high positivity rate. The service meets its primary objective of assisting general practitioners in the management of patients with suspected ischaemic heart disease, and may have helped to optimise resources by reducing the number of referrals to cardiologists. It has helped prioritize patient management and may have benefited high risk patients by facilitating rapid identification and referral. It can be run safely and effectively by trained technicians. PMID- 10455094 TI - Effect of an alpha(2) agonist (mivazerol) on limiting myocardial ischaemia in stable angina. AB - A specific alpha(2) agonist, mivazerol, known to be effective in reducing myocardial ischaemia when given intravenously immediately before an exercise tolerance test, produced a significant increase in exercise duration and time to the onset of angina when given orally over a two week period to 25 patients with stable angina. A non-significant trend to reduction in electrocardiographic signs of ischaemia was also noted. The clinical relevance of this improvement now needs to be tested in larger numbers. PMID- 10455095 TI - A variant of long QT syndrome manifested as fetal tachycardia and associated with ventricular septal defect. AB - Two patients with a novel variant of long QT syndrome are described. The clinical course was characterised by an in utero onset of ventricular tachycardia and atrioventricular block (at 26 and 30 weeks' gestational age, respectively), and an association with a ventricular septal defect. Studies of both patients' families identified relatives with prolonged QT interval, syncope, or sudden death. One patient died of intractable ventricular tachycardia at 4 days old. The other received beta blocker treatment and a pacemaker. She died suddenly at the age of 10 months. The unique association with ventricular septal defect and the malignant clinical course warrants further molecular diagnosis of this novel variant of long QT syndrome. PMID- 10455096 TI - Restrictive pericarditis. AB - BACKGROUND: Pericardial thickening is an uncommon complication of cardiac surgery. OBJECTIVES: To study pericardial thickening as the cause of severe postoperative venous congestion. SUBJECTS: Two men, one with severe aortic stenosis and single coronary artery disease, and one with coronary artery disease after an old inferior infarction. Both had coronary artery bypass grafting surgery. METHODS: Magnetic resonance imaging (MRI), Doppler echocardiography, and cardiac catheterisation. RESULTS: Venous pressure was raised in both patients. MRI showed mildly thickened pericardium, and cardiac catheterisation indicated diastolic equalization of pressures in the four chambers. Jugular venous pulse showed a dominant "Y" descent coinciding with early diastolic flow in the superior vena cava, and mitral and tricuspid Doppler forward flow proved restrictive physiology. The clinical background suggested pericardial disease so both patients had pericardiectomy. This proved the pericardium to be thickened; the extent of fibrosis also involved the epicardium. CONCLUSIONS: Although rare, restrictive pericarditis (restrictive ventricular physiology resulting from pericardial disease) should be considered to be a separate diagnostic entity because its pathological basis and treatment are different from intrinsic myocardial disease. This diagnosis may be confirmed by standard investigational techniques or may require diagnostic thoracotomy. PMID- 10455097 TI - Left ventricular pseudoaneurysm complicating infective pericarditis. AB - Cross sectional echocardiography demonstrated a pseudoaneurysm of the left ventricular posterolateral wall close to the atrioventricular junction in a 4 year old girl with infective pericarditis complicating lobar pneumonia. Colour flow Doppler demonstrated bidirectional flow across the communication hole. Surgical resection was successful. PMID- 10455098 TI - Transcatheter closure of ventricular septal defect using detachable steel coil. AB - Transcatheter closure has been attempted in selected cases of ventricular septal defects using Lock clamshell device, Rashkind umbrella device, and Sideris buttoned device. A small perimembranous ventricular septal defect with aneurysm of membranous septum in a 12 year old girl was closed successfully with a detachable steel coil of 8 mm diameter with four loops, after failure of deployment of a Rashkind double umbrella device. PMID- 10455099 TI - Staphylococcus capitis endocarditis: two cases and review of the literature. AB - Coagulase negative staphylococci are the principal cause of prosthetic valve endocarditis but are a rare cause of native valve infections. However, the incidence of native valve endocarditis is increasing. Staphylococcus capitis is a coagulase negative staphylococcus with the capacity to cause endocarditis on native heart valves. Two cases of native valve endocarditis caused by S capitis are presented; both in patients with aortic valve disease. The patients were cured with prolonged intravenous vancomycin and rifampicin and did not need surgery during the acute phase of the illness. Five of the six previously described cases of endocarditis caused by this organism occurred on native valves and responded to medical treatment alone. PMID- 10455100 TI - Hereditary cardiac amyloidosis associated with the transthyretin Ile122 mutation in a white man. AB - An 83 year old white man with atrial fibrillation was admitted to hospital after a cerebral infarct. Echocardiography was characteristic of cardiac amyloid deposition and subsequent tests confirmed amyloidosis of transthyretin (TTR) type, in association with the Ile122 mutation of the TTR gene; this has only been reported previously in African Americans in whom it occurs with an allele frequency of 2%. Haplotype analysis did not suggest a different founder than for the African Ile122 mutation. Cardiac amyloidosis should be considered among elderly patients presenting with cardiac failure and/or arrhythmia, particularly if they are resistant to conventional treatment; if confirmed, it should be followed by precise characterisation of amyloid fibril type. The prevalence of autosomal dominant cardiac TTR amyloidosis in elderly white people is unknown but early diagnosis and supportive treatment may prevent complications among affected family members. PMID- 10455101 TI - Intravascular ultrasound in the diagnosis of the no-reflow phenomenon after primary angioplasty for myocardial infarction. PMID- 10455102 TI - Isoprostanes: formation, analysis and use as indices of lipid peroxidation in vivo. PMID- 10455103 TI - Calmodulin binds to p21(Cip1) and is involved in the regulation of its nuclear localization. AB - p21(Cip1), first described as an inhibitor of cyclin-dependent kinases, has recently been shown to have a function in the formation of cyclin D-Cdk4 complexes and in their nuclear translocation. The dual behavior of p21(Cip1) may be due to its association with other proteins. Different evidence presented here indicate an in vitro and in vivo interaction of p21(Cip1) with calmodulin: 1) purified p21(Cip1) is able to bind to calmodulin-Sepharose in a Ca(2+)-dependent manner, and this binding is inhibited by the calmodulin-binding domain of calmodulin-dependent kinase II; 2) both molecules coimmunoprecipitate when extracted from cellular lysates; and 3) colocalization of calmodulin and p21(Cip1) can be detected in vivo by electron microscopy immunogold analysis. The carboxyl-terminal domain of p21(Cip1) is responsible for the calmodulin interaction, since p21(145-164) peptide is also able to bind calmodulin and to compete with full-length p21(Cip1) for the calmodulin binding. Because treatment of cells with anti-calmodulin drugs decreases the nuclear accumulation of p21(Cip1), we hypothesize that calmodulin interaction with p21(Cip1) is important for p21(Cip1), and in consequence for cyclin D-Cdk4, translocation into the cell nucleus. PMID- 10455104 TI - GRFbeta, a novel regulator of calcium signaling, is expressed in pancreatic beta cells and brain. AB - By screening for genes expressed differentially in pancreatic beta cells, we have isolated a cDNA encoding GRFbeta, a novel 178-amino acid protein whose N terminus is identical to that of GRF1, a calcium-dependent guanine nucleotide exchange factor, and whose C terminus is unrelated to known proteins. We show that both GRF1 and GRFbeta are expressed selectively in beta cell lines, pancreatic islet cells and brain. Treatment of beta cell lines (betaTC1 and HIT) with calcium ionophore led to a significant elevation in activity of the Ras signal transduction pathway, as determined by phosphorylation of extracellular signal related kinase (ERK). Transfection of beta cells with a plasmid encoding a dominant negative variant of GRF1 led to 70% reduction in ERK phosphorylation, consistent with a role for GRF1 in calcium-dependent Ras signaling in these cells. To examine the possible function of GRFbeta, cultured cells were transfected with a GRFbeta expression vector. This led to a significant reduction in both GRF1-dependent ERK phosphorylation and AP1-dependent reporter gene activity. The results suggest that GRF1 plays a role in mediating calcium dependent signal transduction in beta cells and that GRFbeta represents a novel dominant negative modulator of Ras signaling. PMID- 10455105 TI - Association of neuronal calcium channels with modular adaptor proteins. AB - Presynaptic voltage-gated calcium (Ca(2+)) channels mediate Ca(2+) influx into the presynaptic terminal that triggers synaptic vesicle fusion and neurotransmitter release. The immediate proximity of Ca(2+) channels to the synaptic vesicle release apparatus is critical for rapid and efficient synaptic transmission. In a series of biochemical experiments, we demonstrate a specific association of the cytosolic carboxyl terminus of the N-type Ca(2+) channel pore forming alpha(1B) subunit with the modular adaptor proteins Mint1 and CASK. The carboxyl termini of alpha(1B) bind to the first PDZ domain of Mint1 (Mint1-1). The proline-rich region present in the carboxyl termini of alpha(1B) binds to the SH3 domain of CASK. Mint1-1 is specific for the E/D-X-W-C/S-COOH consensus, which defines a novel class of PDZ domains (class III). The Mint1-1 PDZ domain-binding motif is present only in the "long" carboxyl-terminal splice variants of N-type (alpha(1B)) and P/Q-type (alpha(1A)) Ca(2+) channels, but not in R-type (alpha(1E)) or L-type (alpha(1C)) Ca(2+) channels. Our results directly link presynaptic Ca(2+) channels to a macromolecular complex formed by modular adaptor proteins at synaptic junction and advance our understanding of coupling between cell adhesion and synaptic vesicle exocytosis. PMID- 10455106 TI - Characterization of the product-inhibited complex in catalysis by human manganese superoxide dismutase. AB - The reduction with excess H(2)O(2) of human Mn(III) superoxide dismutase (SOD) and the active-site mutant Y34F Mn(III)SOD was measured by scanning stopped-flow spectrophotometry and revealed the presence of an intermediate in the reduction of the manganese. The visible absorption spectrum of this intermediate closely resembled that of the enzyme in the inhibited, zero-order phase of the catalyzed disproportionation of superoxide. The decay of the visible spectrum of this intermediate was 2-fold faster for the wild-type compared with the mutant Y34F Mn SOD. This correlates with the enhanced product inhibition of Y34F during the catalysis of O-(2) dismutation. The visible spectrum of the product-inhibited complex resembles that of the azide-Mn-SOD complex, suggesting that the inhibited complex has expanded geometry about the metal to octahedral. This study shows that the inhibited complex responsible for the zero-order phase in the catalysis by Mn-SOD of superoxide dismutation can be reached through both the forward (O (2)) and reverse (H(2)O(2)) reactions, supporting a mechanism in which the zero order phase results from product inhibition. PMID- 10455107 TI - MCD encodes peroxisomal and cytoplasmic forms of malonyl-CoA decarboxylase and is mutated in malonyl-CoA decarboxylase deficiency. AB - Malonyl-CoA decarboxylase (MCD) catalyzes the proton-consuming conversion of malonyl-CoA to acetyl-CoA and CO(2). Although defects in MCD activity are associated with malonyl-CoA decarboxylase deficiency, a lethal disorder characterized by cardiomyopathy and developmental delay, the metabolic role of this enzyme in mammals is unknown. A computer-based search for novel peroxisomal proteins led to the identification of a candidate gene for human MCD, which encodes a protein with a canonical type-1 peroxisomal targeting signal of serine lysine-leucine(COOH). We observed that recombinant MCD protein has high intrinsic malonyl-CoA decarboxylase activity and that a malonyl-CoA decarboxylase-deficient patient has a severe mutation in the MCD gene (c.947-948delTT), confirming that this gene encodes human MCD. Subcellular fractionation experiments revealed that MCD resides in both the cytoplasm and peroxisomes. Cytoplasmic MCD is positioned to play a role in the regulation of cytoplasmic malonyl-CoA abundance and, thus, of mitochondrial fatty acid uptake and oxidation. This hypothesis is supported by the fact that malonyl-CoA decarboxylase-deficient patients display a number of phenotypes that are reminiscent of mitochondrial fatty acid oxidation disorders. Additional support for this hypothesis comes from our observation that MCD mRNA is most abundant in cardiac and skeletal muscles, tissues in which cytoplasmic malonyl-CoA is a potent inhibitor of mitochondrial fatty acid oxidation and which derive significant amounts of energy from fatty acid oxidation. As for the role of peroxisomal MCD, we propose that this enzyme may be involved in degrading intraperoxisomal malonyl-CoA, which is generated by the peroxisomal beta oxidation of odd chain-length dicarboxylic fatty acids. PMID- 10455108 TI - Engagement of Gab1 and Gab2 in erythropoietin signaling. AB - Several signaling cascades are activated during engagement of the erythropoietin receptor to mediate the biological effects of erythropoietin. The members of the insulin receptor substrate (IRS) family of proteins play a central role in signaling for various growth factor receptors and cytokines by acting as docking proteins for the SH2 domains of signaling elements, linking cytokine receptors to diverse downstream pathways. In the present study we provide evidence that the recently cloned IRS-related proteins, Gab1 and Gab2, of the Gab family of proteins, are rapidly phosphorylated on tyrosine during erythropoietin treatment of erythropoietin-responsive cells and provide docking sites for the engagement of the SHP2 phosphatase and the p85 subunit of the phosphatidylinositol 3' kinase. Furthermore, our data show that Gab1 is the primary IRS-related protein activated by erythropoietin in primary erythroid progenitor cells. In studies to identify the erythropoietin receptor domains required for activation of Gab proteins, we found that tyrosines 425 and 367 in the cytoplasmic domain of the erythropoietin receptor are required for the phosphorylation of Gab2. Taken together, our data demonstrate that Gab proteins are engaged in erythropoietin signaling to mediate downstream activation of the SHP2 and phosphatidylinositol 3'-kinase pathways and possibly participate in the generation of the erythropoietin-induced mitogenic responses. PMID- 10455109 TI - Identification of amino acid residues responsible for the pyrimidine and purine nucleoside specificities of human concentrative Na(+) nucleoside cotransporters hCNT1 and hCNT2. AB - hCNT1 and hCNT2 mediate concentrative (Na(+)-linked) cellular uptake of nucleosides and nucleoside drugs by human cells and tissues. The two proteins (650 and 658 residues, 71 kDa) are 72% identical in sequence and contain 13 putative transmembrane helices (TMs). When produced in Xenopus oocytes, recombinant hCNT1 is selective for pyrimidine nucleosides (system cit), whereas hCNT2 is selective for purine nucleosides (system cif). Both transport uridine. We have used (i) chimeric constructs between hCNT1 and hCNT2, (ii) sequence comparisons with a newly identified broad specificity concentrative nucleoside transporter (system cib) from Eptatretus stouti, the Pacific hagfish (hfCNT), and (iii) site-directed mutagenesis of hCNT1 to identify two sets of adjacent residues in TMs 7 and 8 of hCNT1 (Ser(319)/Gln(320) and Ser(353)/Leu(354)) that, when converted to the corresponding residues in hCNT2 (Gly(313)/Met(314) and Thr(347)/Val(348)), changed the specificity of the transporter from cit to cif. Mutation of Ser(319) in TM 7 of hCNT1 to Gly enabled transport of purine nucleosides, whereas concurrent mutation of Gln(320) to Met (which had no effect on its own) augmented this transport. The additional mutation of Ser(353) to Thr in TM 8 converted hCNT1/S319G/Q320M, from cib to cif, but with relatively low adenosine transport activity. Additional mutation of Leu(354) to Val (which had no effect on its own) increased the adenosine transport capability of hCNT1/S319G/Q320M/S353T, producing a full cif-type transporter phenotype. On its own, the S353T mutation converted hCNT1 into a transporter with novel uridine selective transport properties. Helix modeling of hCNT1 placed Ser(319) (TM 7) and Ser(353) (TM 8) within the putative substrate translocation channel, whereas Gln(320) (TM 7) and Leu(354) (TM 8) may exert their effects through altered helix packing. PMID- 10455110 TI - The carboxyl terminus of the bacteriophage T4 DNA polymerase contacts its sliding clamp at the subunit interface. AB - The location of the interaction of the COOH terminus of the bacteriophage T4 DNA polymerase with its trimeric, circular sliding clamp has been established. A peptide corresponding to the COOH terminus of the DNA polymerase was labeled with a fluorophore and fluorescence spectroscopy used to show that it forms a specific complex with the sliding clamp by virtue of its low K(D) value (7.1 +/- 1.0 microM). The same peptide was labeled with a photoaffinity probe and cross-linked to the sliding clamp. Mass spectrometry of tryptic digests determined the sole linkage point to be Ala-159 on the sliding clamp, an amino acid that lies on the subunit interface. These results demonstrate that the COOH terminus of the DNA polymerase is inserted into the subunit interface of its sliding clamp, thereby conferring processivity to the DNA polymerase. PMID- 10455111 TI - The heme complex of Hmu O, a bacterial heme degradation enzyme from Corynebacterium diphtheriae. Structure of the catalytic site. AB - Hmu O, a heme degradation enzyme in Corynebacterium diphtheriae, forms a stoichiometric complex with iron protoporphyrin IX and catalyzes the oxygen dependent conversion of hemin to biliverdin, carbon monoxide, and free iron. Using a multitude of spectroscopic techniques, we have determined the axial ligand coordination of the heme-Hmu O complex. The ferric complex shows a pH dependent reversible transition between a water-bound hexacoordinate high spin neutral pH form and an alkaline form, having high spin and low spin states, with a pK(a) of 9. (1)H NMR, EPR, and resonance Raman of the heme-Hmu O complex establish that a neutral imidazole of a histidine residue is the proximal ligand of the complex, similar to mammalian heme oxygenase. EPR of the deoxy cobalt porphyrin IX-Hmu O complex confirms this proximal histidine coordination. Oxy cobalt-Hmu O EPR reveals a hydrogen-bonding interaction between the O(2) and an exchangeable proton in the Hmu O distal pocket and two distinct orientations for the bound O(2). Mammalian heme oxygenase has only one O(2) orientation. This difference and the mixed spin states at alkaline pH indicate structural differences in the distal environment between Hmu O and its mammalian counterpart. PMID- 10455112 TI - Inhibition and restoration of prolactin signal transduction by suppressors of cytokine signaling. AB - Prolactin (PRL) has been shown to activate the cytoplasmic tyrosine kinase Janus kinase 2 (Jak2) and the subsequent recruitment of various signaling molecules including members of the signal transducer and activator of transcription family of transcription factors. Recently, an expanding family of cytokine-inducible inhibitors of signaling has been identified that initially included four members: suppressor of cytokine signaling (SOCS)-1, SOCS-2, SOCS-3, and cytokine-inducible src homology domain 2 (SH-2) proteins. The present study analyzes the role of these members in PRL signaling. Constitutive expression of SOCS-1 and SOCS-3 suppressed PRL-induced signal transducer and activator of transcription 5 dependent gene transcription, and Jak2 tyrosine kinase activity was greatly reduced in the presence of SOCS-1 or SOCS-3. SOCS-1 was shown to associate with Jak2, whereas SOCS-2 was associated with the prolactin receptor. Co-transfection studies were conducted to further analyze the interactions of SOCS proteins. SOCS 2 was shown to suppress the inhibitory effect of SOCS-1 by restoring Jak2 kinase activity but did not affect the inhibitory effect of SOCS-3 on PRL signaling. Northern blot analysis revealed that SOCS-3 and SOCS-1 genes were transiently expressed in response to PRL, both in vivo and in vitro, whereas the expression of SOCS-2 and CIS genes was still elevated 24 h after hormonal stimulation. We thus propose that the early expressed SOCS genes (SOCS-1 and SOCS-3) switch off PRL signaling and that the later expressed SOCS-2 gene can restore the sensitivity of cells to PRL, partly by suppressing the SOCS-1 inhibitory effect. PMID- 10455113 TI - alpha-oxidation of fatty acids in higher plants. Identification of a pathogen inducible oxygenase (piox) as an alpha-dioxygenase and biosynthesis of 2 hydroperoxylinolenic acid. AB - A pathogen-inducible oxygenase in tobacco leaves and a homologous enzyme from Arabidopsis were recently characterized (Sanz, A., Moreno, J. I., and Castresana, C. (1998) Plant Cell 10, 1523-1537). Linolenic acid incubated at 23 degrees C with preparations containing the recombinant enzymes underwent alpha-oxidation with the formation of a chain-shortened aldehyde, i.e., 8(Z),11(Z), 14(Z) heptadecatrienal (83%), an alpha-hydroxy acid, 2(R)-hydroxy-9(Z),12(Z),15(Z) octadecatrienoic acid (15%), and a chain-shortened fatty acid, 8(Z),11(Z),14(Z) heptadecatrienoic acid (2%). When incubations were performed at 0 degrees C, 2(R) hydroperoxy-9(Z),12(Z),15(Z)-octadecatrienoic acid was obtained as the main product. An intermediary role of 2(R)-hydroperoxy-9(Z), 12(Z),15(Z) octadecatrienoic acid in alpha-oxidation was demonstrated by re-incubation experiments, in which the hydroperoxide was converted into the same alpha oxidation products as those formed from linolenic acid. 2(R)-Hydroperoxy 9(Z),12(Z), 15(Z)-octadecatrienoic acid was chemically unstable and had a half life time in buffer of about 30 min at 23 degrees C. Extracts of cells expressing the recombinant oxygenases accelerated breakdown of the hydroperoxide (half-life time, about 3 min at 23 degrees C), however, this was not attributable to the recombinant enzymes since the same rate of hydroperoxide degradation was observed in the presence of control cells not expressing the enzymes. No significant discrimination between enantiomers was observed in the degradation of 2(R,S) hydroperoxy-9(Z)-octadecenoic acid in the presence of recombinant oxygenases. A previously studied system for alpha-oxidation in cucumber was re-examined using the newly developed techniques and was found to catalyze the same conversions as those observed with the recombinant enzymes, i.e. enzymatic alpha-dioxygenation of fatty acids into 2(R)-hydroperoxides and a first order, non-stereoselective degradation of hydroperoxides into alpha-oxidation products. It was concluded that the recombinant enzymes from tobacco and Arabidopsis were both alpha dioxygenases, and that members of this new class of enzymes catalyze the first step of alpha-oxidation in plant tissue. PMID- 10455114 TI - Alternatives to the isomerase-dependent pathway for the beta-oxidation of oleic acid are dispensable in Saccharomyces cerevisiae. Identification of YOR180c/DCI1 encoding peroxisomal delta(3,5)-delta(2,4)-dienoyl-CoA isomerase. AB - Fatty acids with double bonds at odd-numbered positions such as oleic acid can enter beta-oxidation via a pathway relying solely on the auxiliary enzyme Delta(3)-Delta(2)-enoyl-CoA isomerase, termed the isomerase-dependent pathway. Two novel alternative pathways have recently been postulated to exist in mammals, and these additionally depend on Delta(3,5)-Delta(2,4)-dienoyl-CoA isomerase (di isomerase-dependent) or on Delta(3,5)-Delta(2,4)-dienoyl-CoA isomerase and 2,4 dienoyl-CoA reductase (reductase-dependent). We report the identification of the Saccharomyces cerevisiae oleic acid-inducible DCI1 (YOR180c) gene encoding peroxisomal di-isomerase. Enzyme assays conducted on soluble extracts derived from yeast cells overproducing Dci1p using 3,5,8,11,14-eicosapentenoyl-CoA as substrate demonstrated a specific di-isomerase activity of 6 nmol x min(-1) per mg of protein. Similarly enriched extracts from eci1Delta cells lacking peroxisomal 3,2-isomerase additionally contained an intrinsic 3,2-isomerase activity that could generate 3, 5,8,11,14-eicosapentenoyl-CoA from 2,5,8,11,14 eicosapentenoyl-CoA but not metabolize trans-3-hexenoyl-CoA. Amplification of this intrinsic activity replaced Eci1p since it restored growth of the eci1Delta strain on petroselinic acid for which di-isomerase is not required whereas Eci1p is. Heterologous expression in yeast of rat di-isomerase resulted in a peroxisomal protein that was enzymatically active but did not re-establish growth of the eci1Delta mutant on oleic acid. A strain devoid of Dci1p grew on oleic acid to wild-type levels, whereas one lacking both Eci1p and Dci1p grew as poorly as the eci1Delta mutant. Hence, we reasoned that yeast di-isomerase does not additionally represent a physiological 3,2-isomerase and that Dci1p and the postulated alternative pathways in which it is entrained are dispensable for degrading oleic acid. PMID- 10455115 TI - Redox regulation of cell signaling by selenocysteine in mammalian thioredoxin reductases. AB - The intracellular generation of reactive oxygen species, together with the thioredoxin and glutathione systems, is thought to participate in redox signaling in mammalian cells. The activity of thioredoxin is dependent on the redox status of thioredoxin reductase (TR), the activity of which in turn is dependent on a selenocysteine residue. Two mammalian TR isozymes (TR2 and TR3), in addition to that previously characterized (TR1), have now been identified in humans and mice. All three TR isozymes contain a selenocysteine residue that is located in the penultimate position at the carboxyl terminus and which is encoded by a UGA codon. The generation of reactive oxygen species in a human carcinoma cell line was shown to result in both the oxidation of the selenocysteine in TR1 and a subsequent increase in the expression of this enzyme. These observations identify the carboxyl-terminal selenocysteine of TR1 as a cellular redox sensor and support an essential role for mammalian TR isozymes in redox-regulated cell signaling. PMID- 10455116 TI - Functional analysis of paralogous thiol-disulfide oxidoreductases in Bacillus subtilis. AB - The in vivo formation of disulfide bonds, which is critical for the stability and/or activity of many proteins, is catalyzed by thiol-disulfide oxidoreductases. In the present studies, we show that the Gram-positive eubacterium Bacillus subtilis contains three genes, denoted bdbA, bdbB, and bdbC, for thiol-disulfide oxidoreductases. Escherichia coli alkaline phosphatase, containing two disulfide bonds, was unstable when secreted by B. subtilis cells lacking BdbB or BdbC, and notably, the expression levels of bdbB and bdbC appeared to set a limit for the secretion of active alkaline phosphatase. Cells lacking BdbC also showed decreased stability of cell-associated forms of E. coli TEM-beta-lactamase, containing one disulfide bond. In contrast, BdbA was not required for the stability of alkaline phosphatase or beta-lactamase. Because BdbB and BdbC are typical membrane proteins, our findings suggest that they promote protein folding at the membrane-cell wall interface. Interestingly, pre beta-lactamase processing to its mature form was stimulated in cells lacking BdbC, suggesting that the unfolded form of this precursor is a preferred substrate for signal peptidase. Surprisingly, cells lacking BdbC did not develop competence for DNA uptake, indicating the involvement of disulfide bond containing proteins in this process. Unlike E. coli and yeast, none of the thiol disulfide oxidoreductases of B. subtilis was required for growth in the presence of reducing agents. In conclusion, our observations indicate that BdbB and BdbC have a general role in disulfide bond formation, whereas BdbA may be dedicated to a specific process. PMID- 10455117 TI - Adventures in membrane protein topology. A study of the membrane-bound state of colicin E1. AB - The molecular aggregate size of the closed state of the colicin E1 channel was determined by fluorescence resonance energy transfer experiments involving a fluorescence donor (three tryptophans, wild-type protein) and a fluorescence acceptor (5-(((acetyl)amino)ethyl)aminonaphthalene-1-sulfonic acid (AEDANS), Trp deficient protein). There was no evidence of energy transfer between the donor and acceptor species when bound to membrane large unilamellar vesicles. These experiments led to the conclusion that the colicin E1 channel is monomeric in the membrane-bound closed channel state. Experiments were also conducted to study the membrane topology of the closed colicin channel in membrane large unilamellar vesicles using acrylamide as the membrane-impermeant, nonionic quencher of tryptophan fluorescence in a battery of single tryptophan mutant proteins. Furthermore, additional fluorescence parameters, including fluorescence emission maximum, fluorescence quantum yield, and fluorescence decay times, were used to assist in mapping the topology of the closed channel. Results suggest that the closed channel comprises most of the polypeptide of the channel domain and that the hydrophobic anchor domain does not transverse the membrane bilayer but nonetheless is deeply embedded within the hydrocarbon core of the membrane. Finally, a model is proposed which features at least two states that are in rapid equilibrium with each other and in which one state is more heavily populated than the other. PMID- 10455118 TI - Agonist-dependent up-regulation of human somatostatin receptor type 1 requires molecular signals in the cytoplasmic C-tail. AB - We have previously reported that the human somatostatin receptor type 1 (hSSTR1) stably expressed in Chinese hamster ovary-K1 cells does not internalize but instead up-regulates at the membrane during continued agonist treatment (1 microM somatostatin (SST)-14 x 22 h). Here we have investigated the molecular basis of hSSTR1 up-regulation. hSSTR1 was up-regulated by SST in a time-, temperature-, and dose-dependent manner to saturable levels, in intact cells but not in membrane preparations. Although hSSTR1 was acutely desensitized to adenylyl cyclase coupling after 1 h SST-14 treatment, continued agonist exposure (22 h) restored functional effector coupling. Up-regulation was unaffected by cycloheximide but blocked by okadaic acid. Confocal fluorescence immunocytochemistry of intact and permeabilized cells showed progressive, time dependent increase in surface hSSTR1 labeling, associated with depletion of intracellular SSTR1 immunofluorescent vesicles. To investigate the structural domains of hSSTR1 responsible for up-regulation, we constructed C-tail deletion (Delta) mutants and chimeric hSSTR1-hSSTR5 receptors. Human SSTR5 was chosen because it internalizes readily, displays potent C-tail internalization signals, and does not up-regulate. Like wild type hSSTR1, Delta C-tail hSSTR1 did not internalize and additionally lost the ability to up-regulate. Swapping the C-tail of hSSTR1 with that of hSSTR5 induced internalization (27%) but not up regulation. Substitution of hSSTR5 C-tail with that of hSSTR1 converted the chimeric receptor to one resembling wild type hSSTR1 (poor internalization, 71% up-regulation). These results show that ligand-induced up-regulation of hSSTR1 occurs by a temperature-dependent active process of receptor recruitment from a pre-existing cytoplasmic pool to the plasma membrane. It does not require new protein synthesis or signal transduction, is sensitive to dephosphorylation events, and critically dependent on molecular signals in the receptor C-tail. PMID- 10455119 TI - Glucose-6-phosphatase overexpression lowers glucose 6-phosphate and inhibits glycogen synthesis and glycolysis in hepatocytes without affecting glucokinase translocation. Evidence against feedback inhibition of glucokinase. AB - In hepatocytes glucokinase (GK) and glucose-6-phosphatase (Glc-6-Pase)(1) have converse effects on glucose 6-phosphate (and fructose 6-phosphate) levels. To establish whether hexose 6-phosphate regulates GK binding to its regulatory protein, we determined the effects of Glc-6-Pase overexpression on glucose metabolism and GK compartmentation. Glc-6-Pase overexpression (4-fold) decreased glucose 6-phosphate levels by 50% and inhibited glycogen synthesis and glycolysis with a greater negative control coefficient on glycogen synthesis than on glycolysis, but it did not affect the response coefficients of glycogen synthesis or glycolysis to glucose, and it did not increase the control coefficient of GK or cause dissociation of GK from its regulatory protein, indicating that in hepatocytes fructose 6-phosphate does not regulate GK translocation by feedback inhibition. GK overexpression increases glycolysis and glycogen synthesis with a greater control coefficient on glycogen synthesis than on glycolysis. On the basis of the similar relative control coefficients of GK and Glc-6-Pase on glycogen synthesis compared with glycolysis, and the lack of effect of Glc-6-Pase overexpression on GK translocation or the control coefficient of GK, it is concluded that the main regulatory function of Glc-6-Pase is to buffer the glucose 6-phosphate concentration. This is consistent with recent findings that hyperglycemia stimulates Glc-6-Pase gene transcription. PMID- 10455120 TI - Skp, a molecular chaperone of gram-negative bacteria, is required for the formation of soluble periplasmic intermediates of outer membrane proteins. AB - Using a cross-linking approach, we have analyzed the function of Skp, a presumed molecular chaperone of the periplasmic space of Escherichia coli, during the biogenesis of an outer membrane protein (OmpA). Following its transmembrane translocation, OmpA interacts with Skp in close vicinity to the plasma membrane. In vitro, Skp was also found to bind strongly and specifically to pOmpA nascent chains after their release from the ribosome suggesting the ability of Skp to recognize early folding intermediates of outer membrane proteins. Pulse labeling of OmpA in spheroplasts prepared from an skp null mutant revealed a specific requirement of Skp for the release of newly translocated outer membrane proteins from the plasma membrane. Deltaskp mutant cells are viable and show only slight changes in the physiology of their outer membranes. In contrast, double mutants deficient both in Skp and the periplasmic protease DegP (HtrA) do not grow at 37 degrees C in rich medium. We show that in the absence of an active DegP, a lack of Skp leads to the accumulation of protein aggregates in the periplasm. Collectively, our data demonstrate that Skp is a molecular chaperone involved in generating and maintaining the solubility of early folding intermediates of outer membrane proteins in the periplasmic space of Gram-negative bacteria. PMID- 10455121 TI - Dissociation of functional roles of dynamin in receptor-mediated endocytosis and mitogenic signal transduction. AB - Dynamin plays a critical role in the membrane fission mechanism that mediates regulated endocytosis of many G protein-coupled receptors. In addition, dynamin is required for ligand-induced activation of mitogen-activated protein kinase by certain receptors, raising a general question about the role of dynamin in mitogenic signal transduction. Here we report that endocytosis of mu and delta opioid receptors is not required for efficient ligand-induced activation of mitogen-activated protein kinase. Nevertheless, mitogenic signaling mediated by these receptors is specifically dynamin-dependent. Thus a functional role of dynamin in mitogenic signaling can be dissociated from its role in receptor mediated endocytosis, suggesting a previously unidentified and distinct role of dynamin in signal transduction by certain G protein-coupled receptors. PMID- 10455122 TI - Cell-cell dissociation upon epithelial cell scattering requires a step mediated by the proteasome. AB - During development, tissue repair, and tumor metastasis, both cell-cell dissociation and cell migration occur and appear to be intimately linked, such as during epithelial "scattering." Here we show that cell-cell dissociation during scattering induced by hepatocyte growth factor (HGF) or activation of the temperature-sensitive v-Src tyrosine kinase in MDCK cells can be blocked by inhibiting the proteasome with lactacystin and MG132. Although both proteins of the tight junction and the adherens junction redistributed during cell scattering, proteasome inhibitors largely prevented this process, resulting in the stabilization of Triton X-100-insoluble tight junction proteins as well as adherens junction proteins at sites of cell-cell contact. Proteasome inhibition also led to a decrease of E-cadherin turnover in (35)S-labeled cells. In addition, proteasome inhibition partly preserved cell polarity, as determined by the subcellular distribution of Na(+),K(+)-ATPase (basolateral marker) and gp135 (apical marker), and the structure of the subcortical actin ring, both of which are normally disrupted during scattering. However, cells were able to establish focal contacts, and single cell migration toward HGF was unaffected by proteasome inhibition in quantitative assays, indicating that cell-cell dissociation during scattering occurs independently of anchorage-dependent cell migration. Thus, a proteasome-dependent step during scattering induced by HGF and pp60(v-Src) appears to be essential for cell-cell dissociation, disassembly of junctional components, and (at least indirectly) it also plays a role in the loss of protein polarity. PMID- 10455123 TI - Signal peptide peptidase- and ClpP-like proteins of Bacillus subtilis required for efficient translocation and processing of secretory proteins. AB - Signal peptides direct the export of secretory proteins from the cytoplasm. After processing by signal peptidase, they are degraded in the membrane and cytoplasm. The resulting fragments can have signaling functions. These observations suggest important roles for signal peptide peptidases. The present studies show that the Gram-positive eubacterium Bacillus subtilis contains two genes for proteins, denoted SppA and TepA, with similarity to the signal peptide peptidase A of Escherichia coli. Notably, TepA also shows similarity to ClpP proteases. SppA of B. subtilis was only required for efficient processing of pre-proteins under conditions of hyper-secretion. In contrast, TepA depletion had a strong effect on pre-protein translocation across the membrane and subsequent processing, not only under conditions of hyper-secretion. Unlike SppA, which is a typical membrane protein, TepA appears to have a cytosolic localization, which is consistent with the observation that TepA is involved in early stages of the secretion process. Our observations demonstrate that SppA and TepA have a role in protein secretion in B. subtilis. Based on their similarity to known proteases, it seems likely that SppA and TepA are specifically required for the degradation of proteins or (signal) peptides that are inhibitory to protein translocation. PMID- 10455124 TI - Characterization of the carboxyl-terminal domain of the rat glucose-dependent insulinotropic polypeptide (GIP) receptor. A role for serines 426 and 427 in regulating the rate of internalization. AB - Glucose-dependent insulinotropic polypeptide (GIP) is a gastrointestinal hormone involved in the regulation of insulin secretion. In non-insulin-dependent diabetes mellitus insulin responses to GIP are blunted, possibly due to altered signal transduction or reduced receptor number. Site-directed mutagenesis was used to construct truncated GIP receptors to study the importance of the carboxyl terminal tail (CT) in binding, signaling, and receptor internalization. Receptors truncated at amino acids 425, 418, and 405, expressed in COS-7 or CHO-K1 cells, exhibited similar binding to wild type receptors. GIP-dependent cAMP production with the 405 mutant was decreased in COS-7 cells. Maximal cAMP production in CHO K1 cells was reduced with all truncated forms. Binding was undetectable with a receptor truncated at amino acid 400; increasing tail length by adding 5 alanines restored binding and signaling. Mutants produced by alanine scanning of residues 394-401, adjacent to transmembrane domain 7, were all functional. CT truncation by 30 or more amino acids, mutation of serines 426/427, singly or combined, or complete CT serine knockout all reduced receptor internalization rate. The majority of the GIP receptor CT is therefore not required for signaling, a minimum chain length of approximately 405 amino acids is needed for receptor expression, and serines 426 and 427 are important for regulating rate of receptor internalization. PMID- 10455125 TI - Integrin and neurocan binding to L1 involves distinct Ig domains. AB - The cell adhesion molecule L1, a 200-220-kDa type I membrane glycoprotein of the Ig superfamily, mediates many neuronal processes. Originally studied in the nervous system, L1 is expressed by hematopoietic and many epithelial cells, suggesting a more expanded role. L1 supports homophilic L1-L1 and integrin mediated cell binding and can also bind with high affinity to the neural proteoglycan neurocan; however, the binding site is unknown. We have dissected the L1 molecule and investigated the cell binding ability of Ig domains 1 and 6. We report that RGD sites in domain 6 support alpha5beta1- or alphavbeta3-mediated integrin binding and that both RGD sites are essential. Cooperation of RGD sites with neighboring domains are necessary for alpha(5)beta(1). A T cell hybridoma and activated T cells could bind to L1 in the absence of RGDs. This binding was supported by Ig domain 1 and mediated by cell surface-exposed neurocan. Lymphoid and brain-derived neurocan were structurally similar. We also present evidence that a fusion protein of the Ig 1-like domain of L1 can bind to recombinant neurocan. Our results support the notion that L1 provides distinct cell binding sites that may serve in cell-cell or cell-matrix interactions. PMID- 10455126 TI - Electron spin resonance investigation of the cyanyl and azidyl radical formation by cytochrome c oxidase. AB - Cyanide (CN(-)) is a frequently used inhibitor of mitochondrial respiration due to its binding to the ferric heme a(3) of cytochrome c oxidase (CcO). As-isolated CcO oxidized cyanide to the cyanyl radical ((.)CN) that was detected, using the ESR spin-trapping technique, as the 5,5-dimethyl-1-pyrroline N-oxide (DMPO)/(.)CN radical adduct. The enzymatic conversion of cyanide to the cyanyl radical by CcO was time-dependent but not affected by azide (N(3)(-)). The small but variable amounts of compound P present in the as-isolated CcO accounted for this one electron oxidation of cyanide to the cyanyl radical. In contrast, as-isolated CcO exhibited little ability to catalyze the oxidation of azide, presumably because of azide's lower affinity for the CcO. However, the DMPO/(.)N(3) radical adduct was readily detected when H(2)O(2) was included in the system. The results presented here indicate the need to re-evaluate oxidative stress in mitochondria "chemical hypoxia" induced by cyanide or azide to account for the presence of highly reactive free radicals. PMID- 10455127 TI - A pH-dependent conformational change of NhaA Na(+)/H(+) antiporter of Escherichia coli involves loop VIII-IX, plays a role in the pH response of the protein, and is maintained by the pure protein in dodecyl maltoside. AB - Digestion with trypsin of purified His-tagged NhaA in a solution of dodecyl maltoside yields two fragments at alkaline pH but only one fragment at acidic pH. Determination of the amino acid sequence of the N terminus of the cleavage products show that the pH-sensitive cleavage site of NhaA, both in isolated everted membrane vesicles as well as in the pure protein in detergent, is Lys-249 in loop VIII-IX, which connects transmembrane segment VIII to IX. Interestingly, the two polypeptide products of the split antiporter remain complexed and co purify on Ni(2+)-NTA column. Loop VIII-IX has also been found to play a role in the pH regulation of NhaA; three mutations introduced into the loop shift the pH profile of the Na(+)/H(+) antiporter activity as measured in everted membrane vesicles. An insertion mutation introducing Ile-Glu-Gly between residues Lys-249 and Arg-250 (K249-IEG-R250) and Cys replacement of either Val-254 (V254C) or Glu 241 (E241C) cause acidic shift of the pH profile of the antiporter by 0.5, 1, and 0.3 pH units, respectively. Interestingly, the double mutant E241C/V254C introduces a basic shift of more than 1 pH unit with respect to the single mutation V254C. Taken together these results imply the involvement of loop VIII IX in the pH-induced conformational change, which leads to activation of NhaA at alkaline pH. PMID- 10455128 TI - Differential effects of lipopolysaccharide and tumor necrosis factor on monocytic IkappaB kinase signalsome activation and IkappaB proteolysis. AB - The inflammatory mediators lipopolysaccharide (LPS) and tumor necrosis factor (TNF) are potent activators of NF-kappaB. This study compared the effect of these stimuli on endogenous IkappaB kinase (IKK) signalsome activation and IkappaB phosphorylation/proteolysis in human monocytic cells and investigated the role of the signalsome proteins IKK-alpha, IKK-beta, NF-kappaB-inducing kinase (NIK), IKK gamma (NF-kappaB essential modulator), and IKK complex-associated protein. Kinase assays showed that TNF elicited a rapid but short-lived induction of IKK activity with a 3-fold greater effect on IKK-alpha than on IKK-beta, peaking at 5 min. In contrast, LPS predominantly stimulated IKK-beta activity, which slowly increased, peaking at 30 min. A second peak was observed at a later time point following LPS stimulation, which consisted of both IKK-alpha and -beta activity. The endogenous levels of the signalsome components were unaffected by stimulation. Furthermore, our studies showed association of the IKK-alpha/beta heterodimer with NIK, IkappaB-alpha and -epsilon in unstimulated cells. Exposure to LPS or TNF led to differential patterns of IkappaB-alpha and IkappaB-epsilon disappearance from and reassembly with the signalsome, whereas IKK-alpha, IKK-beta, and NIK remained complex-associated. NIK cannot phosphorylate IkappaB-alpha directly, but it appears to be a functionally important subunit, because mutated NIK inhibited stimulus-induced kappaB-dependent transcription more effectively than mutated IKK alpha or -beta. Overexpression of IKK complex-associated protein inhibited stimulus-mediated transcription, whereas NF-kappaB essential modulator enhanced it. The understanding of LPS- and TNF-induced signaling may allow the development of specific strategies to treat sepsis-associated disease. PMID- 10455129 TI - Regulation of insulin-like growth factor-I and -II by glucose in primary cultures of fetal rat hepatocytes. AB - A selective primary culture of fetal rat hepatocytes was established in our laboratory in order to elucidate the molecular mechanisms of action of different factors and conditions on insulin-like growth factor (IGF)-I and -II gene expression during the perinatal period of the rat. In this model we report that, in a serum-free condition and the presence of non-stimulatory doses of insulin, 5 20 mM glucose evoked an increase of IGF-I and -II mRNA abundance. Glucose regulated in a parallel manner IGF peptide secretion, and an excellent correlation was observed between IGF-I and -II mRNA and IGF-I and -II peptide levels in the conditioned media in response to the carbohydrate. The experiment with 2-deoxyglucose suggests that glucose 6-phosphate, but not its further metabolism, is necessary for the induction of IGF transcript abundance in cultured fetal hepatocytes. Finally, the glucose-induced rise in IGF-II mRNA, the main IGF in fetal stages, was mediated by stimulation of gene transcription and increased transcript stability. The results support the idea that IGFs belong to a family of genes that are positively regulated by glucose. PMID- 10455131 TI - Purification and characterization of the serum amyloid A3 enhancer factor. AB - Serum amyloid A (SAA) is a major acute-phase protein synthesized and secreted mainly by the liver. In response to acute inflammation, its expression may be induced up to 1000-fold, primarily as a result of a 200-fold increase in the rate of SAA gene transcription. We showed previously that cytokine-induced transcription of the SAA3 gene promoter requires a transcriptional enhancer that contains three functional elements: two CCAAT/enhancer-binding protein (C/EBP) binding sites and a third site that interacts with a constitutively expressed transcription factor, SAA3 enhancer factor (SEF). Each of these binding sites as well as cooperation among their binding factors is necessary for maximum transcription activation by inflammatory cytokines. Deletion or site-specific mutations in the SEF-binding site drastically reduced SAA3 promoter activity, strongly suggesting that SEF is important in SAA3 promoter function. To further elucidate its role in the regulation of the SAA3 gene, we purified SEF from HeLa nuclear extracts to near homogeneity by using conventional liquid chromatography and DNA affinity chromatography. Ultraviolet cross-linking and Southwestern experiments indicated that SEF consisted of a single polypeptide with an apparent molecular mass of 65 kDa. Protein sequencing and antibody supershift experiments identified SEF as transcription factor LBP-1c/CP2/LSF. Cotransfection of SEF expression vector with SAA3-luciferase reporter resulted in approximately a 5 fold increase in luciferase activity. Interestingly, interleukin-1 treatment of SEF-transfected cells caused dramatic synergistic activation (31-fold) of the SAA3 promoter. In addition to its role in regulating SAA3 gene expression, we provide evidence that SEF could also bind in a sequence-specific manner to the promoters of the alpha(2)-macroglobulin and Aalpha-fibrinogen genes and to an intronic enhancer of the human Wilm's tumor 1 gene, suggesting a functional role in the regulation of these genes. PMID- 10455130 TI - Expression of core 2 beta-1,6-N-acetylglucosaminyltransferase in a human pancreatic cancer cell line results in altered expression of MUC1 tumor associated epitopes. AB - Many tumor-associated epitopes possess carbohydrate as a key component, and thus changes in the activity of glycosyltransferases could play a role in generating these epitopes. In this report we describe the stable transfection of a human pancreatic adenocarcinoma cell line, Panc1-MUC1, with the cDNA for mucin core 2 GlcNAc-transferase (C2GnT), which creates the core 2 beta-1,6 branch in mucin type glycans. These cells lack endogenous C2GnT activity but express a recombinant human MUC1 cDNA. C2GnT-transfected clones expressing different levels of C2GnT were characterized using monoclonal antibodies CC49, CSLEX-1, and SM-3, which recognize tumor-associated epitopes. Increased C2GnT expression led to greatly diminished expression of the CC49 epitope, which we identified as NeuAcalpha2,6(Galbeta1,3)GalNAcalpha-Ser/Thr in the Panc1-MUC1 cells. This was accompanied by the emergence of the CSLEX-1 epitope, sialyl Lewis x (NeuAcalpha2,3Galbeta1,4(Fucalpha1,3)GlcNAc-R), an important selectin ligand. Despite this, however, the C2GnT transfectants could not bind to selectins. Increased C2GnT expression also led to masking of the SM-3 peptide epitope, which persisted after the removal of sialic acid, further suggesting greater complexity of the core 2-associated O-glycans on MUC1. The results of this study suggest that C2GnT could play a regulatory role in the expression of certain tumor associated epitopes. PMID- 10455132 TI - Chloromethyltetramethylrosamine (Mitotracker Orange) induces the mitochondrial permeability transition and inhibits respiratory complex I. Implications for the mechanism of cytochrome c release. AB - We have investigated the interactions with isolated mitochondria and intact cells of chloromethyltetramethylrosamine (CMTMRos), a probe (Mitotracker Orange) that is increasingly used to monitor the mitochondrial membrane potential (Deltapsi(m)) in situ. CMTMRos binds to isolated mitochondria and undergoes a large fluorescence quenching. Most of the binding is energy-independent and can be substantially reduced by sulfhydryl reagents. A smaller fraction of the probe is able to redistribute across the inner membrane in response to a membrane potential, with further fluorescence quenching. Within minutes, however, this energy-dependent fluorescence quenching spontaneously reverts to the same level obtained by treating mitochondria with the uncoupler carbonylcyanide-p trifluoromethoxyphenyl hydrazone. We show that this event depends on inhibition of the mitochondrial respiratory chain at complex I and on induction of the permeability transition pore by CMTMRos, with concomitant depolarization, swelling, and release of cytochrome c. After staining cells with CMTMRos, depolarization of mitochondria in situ with protonophores is accompanied by changes of CMTMRos fluorescence that range between small and undetectable, depending on the probe concentration. A lasting decrease of cellular CMTMRos fluorescence associated with mitochondria only results from treatment with thiol reagents, suggesting that CMTMRos binding to mitochondria in living cells largely occurs at SH groups via the probe chloromethyl moiety irrespective of the magnitude of Deltapsi(m). Induction of the permeability transition precludes the use of CMTMRos as a reliable probe of Deltapsi(m) in situ and demands a reassessment of the conclusion that cytochrome c release can occur without membrane depolarization and/or onset of the permeability transition. PMID- 10455133 TI - Reaction of peroxynitrite with reduced nicotinamide nucleotides, the formation of hydrogen peroxide. AB - NAD(P)H acts as a two-electron reductant in physiological, enzyme-controlled processes. Under nonenzymatic conditions, a couple of one-electron oxidants easily oxidize NADH to the NAD(.) radical. This radical reduces molecular oxygen to the superoxide radical (O-(2)) at a near to the diffusion-controlled rate, thereby subsequently forming hydrogen peroxide (H(2)O(2)). Because peroxynitrite can act as a one-electron oxidant, the reaction of NAD(P)H with both authentic peroxynitrite and the nitric oxide ((. )NO) and O-(2) releasing compound 3 morpholinosydnonimine N-ethylcarbamide (SIN-1) was studied. Authentic peroxynitrite oxidized NADH with an efficiency of approximately 25 and 8% in the absence and presence of bicarbonate/carbon dioxide (HCO(3)(-)/CO(2)), respectively. NADH reacted 5-100 times faster with peroxynitrite than do the known peroxynitrite scavengers glutathione, cysteine, and tryptophan. Furthermore, NADH was found to be highly effective in suppressing peroxynitrite mediated nitration reactions even in the presence of HCO(3)(-)/CO(2). Reaction of NADH with authentic peroxynitrite resulted in the formation of NAD(+) and O-(2) and, thus, of H(2)O(2) with yields of about 3 and 10% relative to the added amounts of peroxynitrite and NADH, respectively. Peroxynitrite generated in situ from SIN-1 gave virtually the same results; however, two remarkable exceptions were recognized. First, the efficiency of NADH oxidation increased to 60-90% regardless of the presence of HCO(3)(-)/CO(2), along with an increase of H(2)O(2) formation to about 23 and 35% relative to the amounts of added SIN-1 and NADH. Second, and more interesting, the peroxynitrite scavenger glutathione (GSH) was needed in a 75-fold surplus to inhibit the SIN-1-dependent oxidation of NADH half maximal in the presence of HCO(3)(-)/CO(2). Similar results were obtained with NADPH. Hence, peroxynitrite or radicals derived from it (such as, e.g. the bicarbonate radical or nitrogen dioxide) indeed oxidize NADH, leading to the formation of NAD(+) and, via O-(2), of H(2)O(2). When peroxynitrite is generated in situ in the presence of HCO(3)(-)/CO(2), i.e. under conditions mimicking the in vivo situation, NAD(P)H effectively competes with other known scavengers of peroxynitrite. PMID- 10455134 TI - AML1 (CBFalpha2) cooperates with B cell-specific activating protein (BSAP/PAX5) in activation of the B cell-specific BLK gene promoter. AB - AML1 plays a critical role during hematopoiesis and chromosomal translocations involving AML1 are commonly associated with different forms of leukemia, including pre-B acute lymphoblastic leukemia. To understand the function of AML1 during B cell differentiation, we analyzed regulatory regions of B cell-specific genes for potential AML1-binding sites and have identified a putative AML1 binding site in the promoter of the B cell-specific tyrosine kinase gene, blk. Gel mobility shift assays and transient transfection assays demonstrate that AML1 binds specifically to this site in the blk promoter and this binding site is important for blk promoter activity. Furthermore, in vitro binding analysis revealed that the AML1 runt DNA-binding domain physically interacts with the paired DNA-binding domain of BSAP, a B cell-specific transcription factor. BSAP has been shown previously to be important for B cell-specific regulation of the blk gene. Physical interaction of AML1 with BSAP correlates with functional cooperativity in transfection studies where AML1 and BSAP synergistically activate blk promoter transcription by more than 50-fold. These results demonstrate physical and functional interactions between AML1 and BSAP and suggest that AML1 is an important factor for regulating a critical B cell specific gene, blk. PMID- 10455135 TI - Identification of wortmannin-sensitive targets in 3T3-L1 adipocytes. DissociationoOf insulin-stimulated glucose uptake and glut4 translocation. AB - The current studies investigated the contribution of phosphatidylinositol 3 kinase (PI3-kinase) isoforms to insulin-stimulated glucose uptake and glucose transporter 4 (GLUT4) translocation. Experiments involving the microinjection of antibodies specific for the p110 catalytic subunit of class I PI3-kinases demonstrated an absolute requirement for this form of the enzyme in GLUT4 translocation. This finding was confirmed by the demonstration that the PI3 kinase antagonist wortmannin inhibits GLUT4 and insulin-responsive aminopeptidase translocation with a dose response identical to that required to inhibit another class I PI3-kinase-dependent event, activation of pp70 S6-kinase. Interestingly, wortmannin inhibited insulin-stimulated glucose uptake at much lower doses, suggesting the existence of a second, higher affinity target of the drug. Subsequent removal of wortmannin from the media shifted this dose-response curve to one resembling that for GLUT4 translocation and pp70 S6-kinase. This is consistent with the lower affinity target being p110, which is irreversibly inhibited by wortmannin. Wortmannin did not reduce glucose uptake in cells stably expressing Myr-Akt, which constitutively induced GLUT4 translocation to the plasma membrane; this demonstrates that wortmannin does not inhibit the transporters directly. In addition to elucidating a second wortmannin-sensitive pathway in 3T3-L1 adipocytes, these studies suggest that the presence of GLUT4 on the plasma membrane is not sufficient for activation of glucose uptake. PMID- 10455136 TI - Mannose 6-Phosphate/Insulin-like growth factor II receptor mediates internalization and degradation of leukemia inhibitory factor but not signal transduction. AB - Leukemia inhibitory factor (LIF) is a multifunctional cytokine belonging to the interleukin-6 subfamily of helical cytokines, all of which use the glycoprotein (gp) 130 subunit for signal transduction. The specific receptor for LIF, gp190, binds this cytokine with low affinity and is also required for signal transduction. We have recently reported that glycosylated LIF produced by transfected Chinese hamster ovary cells also binds to a lectin-like receptor, mannose 6-phosphate/insulin-like growth factor II receptor (Man-6-P/IGFII-R) (Blanchard, F., Raher, S., Duplomb, L., Vusio, P., Pitard, V., Taupin, J. L., Moreau, J. F., Hoflack, B., Minvielle, S., Jacques, Y., and Godard, A. (1998) J. Biol. Chem. 273, 20886-20893). The present study shows that (i) mannose 6 phosphate-containing LIF is naturally produced by a number of normal and tumor cell lines; (ii) other cytokines in the interleukin-6 family do not bind to Man-6 P/IGFII-R; and (iii) another unrelated cytokine, macrophage-colony-stimulating factor, is also able to bind to Man-6-P/IGFII-R in a mannose 6-phosphate sensitive manner. No functional effects or signal transductions mediated by this lectin-like receptor were observed in various biological assays after LIF binding, and mannose 6-phosphate-containing LIF was as active as non-glycosylated LIF. However, mannose 6-phosphate-sensitive LIF binding resulted in rapid internalization and degradation of the cytokine on numerous cell lines, which suggests that Man-6-P/IGFII-R plays an important role in regulating the amounts of LIF available in vivo. PMID- 10455137 TI - Geminate recombination of nitric oxide to endothelial nitric-oxide synthase and mechanistic implications. AB - The nitric-oxide synthase (NOS) catalyzes the oxidation of L-arginine to L citrulline and NO through consumption of oxygen bound to the heme. Because NO is produced close to the heme and may bind to it, its subsequent role in a regulatory mechanism should be scrutinized. We therefore examined the kinetics of NO rebinding after photodissociation in the heme pocket of human endothelial NOS by means of time-resolved absorption spectroscopy. We show that geminate recombination of NO indeed occurs and that this process is strongly modulated by L-Arg. This NO rebinding occurs in a multiphasic fashion and spans over 3 orders of magnitude. In both ferric and ferrous states of the heme, a fast nonexponential picosecond geminate rebinding first takes place followed by a slower nanosecond phase. The rates of both phases decreased, whereas their relative amplitudes are changed by the presence of L-Arg; the overall effect is a slow down of NO rebinding. For the isolated oxygenase domain, the picosecond rate is unchanged, but the relative amplitude of the nanosecond binding decreased. We assigned the nanosecond kinetic component to the rebinding of NO that is still located in the protein core but not in the heme pocket. The implications for a mechanism of regulation involving NO binding are discussed. PMID- 10455138 TI - Activation and routing of membrane-tethered prohormone convertases 1 and 2. AB - Many peptide hormones and neuropeptides are processed by members of the subtilisin-like family of prohormone convertases (PCs), which are either soluble or integral membrane proteins. PC1 and PC2 are soluble PCs that are primarily localized to large dense core vesicles in neurons and endocrine cells. We examined whether PC1 and PC2 were active when expressed as membrane-tethered proteins, and how tethering to membranes alters the biosynthesis, enzymatic activity, and intracellular routing of these PCs. PC1 and PC2 chimeras were constructed using the transmembrane domain and cytoplasmic domain of the amidating enzyme, peptidylglycine alpha-amidating monooxygenase (PAM). The membrane-tethered PCs were rerouted from large dense core vesicles to the Golgi region. In addition, the chimeras were transiently expressed at the cell surface and rapidly internalized to the Golgi region in a fashion similar to PAM. Membrane-tethered PC1 and PC2 exhibited changes in pro-domain maturation rates, N glycosylation, and in the pH and calcium optima required for maximal enzymatic activity against a fluorogenic substrate. In addition, the PC chimeras efficiently cleaved endogenous pro-opiomelanocortin to the correct bioactive peptides. The PAM transmembrane domain/cytoplasmic domain also prevented stimulated secretion of pro-opiomelanocortin products in AtT-20 cells. PMID- 10455139 TI - Sterol regulatory element-binding protein negatively regulates microsomal triglyceride transfer protein gene transcription. AB - We herein report that mRNA expression of microsomal triglyceride transfer protein (MTP) and its protein synthesis decline in response to sterol depletion in HepG2 cells, and we functionally characterized the MTP gene promoter in an effort to investigate the molecular mechanisms by which MTP gene transcription is regulated. Luciferase assays using truncated versions of the reporter gene revealed that the region at -124 to +33 base pairs of the human promoter contains the elements required for the suppression of transcription by sterol depletion. Enforced expression of an active form of sterol regulatory element-binding protein (SREBP)-1 (amino acids 1-487) or -2 (amino acids 1-481), both of which are activated under sterol-depleted conditions, is able to mimic sterol-mediated down-regulation. Either further truncation of the promoter region or mutation of the putative SREBP-binding sequence (5'-GCAGCCCAC-3', -124 to -116 base pairs) abolishes the sterol- and SREBP-dependent transcriptional regulation. Gel mobility shift assay showed that recombinant SREBP-2-(1-481) is able to bind the sequence. Enforced expression of a truncated form of SREBP-2 (amino acids 31 481), which acts as an inhibitor of transcription of the low density lipoprotein receptor gene because it lacks the transcriptional activation domain, also diminishes the luciferase activity, suggesting that direct binding to the promoter region might be sufficient and that the mechanism by which SREBPs inhibit MTP gene expression is distinct from that for the transcriptional stimulation of sterol-regulated genes. Although the SREBP-binding site overlaps a negative insulin-responsive element, insulin negatively regulates MTP gene expression even when the amount of the active form of SREBPs is quite low under the sterol-loaded conditions, indicating that SREBPs only slightly mediate, if at all, the insulin effects. Overall, we conclude that SREBPs are responsible for regulation of lipoprotein secretion via their control of MTP gene expression. Moreover, our results describe for the first time a novel mechanism by which SREBPs negatively regulate expression of the gene encoding the protein involved in lipid metabolism. PMID- 10455140 TI - A conserved amino acid motif (R-X-G-R-R) in the Glut1 glucose transporter is an important determinant of membrane topology. AB - The Glut1 glucose transporter is one of over 300 members of the major facilitator superfamily of membrane transporters. These proteins are extremely diverse in substrate specificity and differ in their transport mechanisms. The two most common features shared by many members of this superfamily are the presence of 12 predicted transmembrane segments and an amino acid motif, R-X-G-R-R, present at equivalent positions within the cytoplasmic loops joining transmembrane segments 2-3 and 8-9. The structural and functional roles of the arginine residues within these motifs in Glut1 were investigated by expression of site-directed mutant transporters in Xenopus oocytes followed by analyses of intrinsic transport activity and the membrane topology of mutant glycosylation-scanning reporter Glut1 molecules. Substitution of lysine residues for the cluster of 3 arginine residues in each of the 2 cytoplasmic pentameric motifs of Glut1 revealed no absolute requirement for arginine side chains at any of the 6 positions for transport of 2-deoxyglucose. However, removal of the 3 positive charges at either site by substitution of glycines for the arginines completely abolished transport activity as the result of a local perturbation in the membrane topology in which the cytoplasmic loop was aberrantly translocated into the exoplasm along with the two flanking transmembrane segments. Substitution of lysines for the arginines had no affect on membrane topology. We conclude that the positive charges in the R-X-G-R-R motif form critical local cytoplasmic anchor points involved in determining the membrane topology of Glut1. These data provide a simple explanation for the presence of this conserved amino acid motif in hundreds of functionally diverse membrane transporters that share a common predicted membrane topology. PMID- 10455141 TI - Cloning and characterization of mouse deoxyguanosine kinase. Evidence for a cytoplasmic isoform. AB - Deoxyguanosine kinase (dGK) is a nuclear gene product that catalyzes the phosphorylation of purine deoxyribonucleosides and their analogues. The human enzyme is located predominantly in the mitochondria, as shown by biochemical fractionation studies and in situ localization of the overexpressed recombinant protein. Here we describe the cloning of mouse dGK cDNA and the identification of a novel amino-terminally truncated isoform that corresponds to about 14% of the total dGK mRNA population in mouse spleen. In situ fluorescence assays suggest that the new isoform cannot translocate into the mitochondria and thus may represent a cytoplasmic enzyme. Expression of mouse dGK mRNA was highly tissue specific and differed from the tissue distribution observed in humans. Recombinant mouse dGK showed similar specific activity and substrate specificity as compared with the human enzyme. The broad specificity, restricted tissue distribution, and location of mouse dGK in multiple cellular compartments raise new considerations with respect to the role of the individual deoxynucleoside kinases in nucleotide metabolism. PMID- 10455142 TI - Osmotic stress inhibits p70/85 S6 kinase through activation of a protein phosphatase. AB - While studying the stress regulation of p70/85 S6 kinase (S6K), we observed that anisomycin and UV light stimulated S6K activity, but that sorbitol inactivated S6K. Pretreatment with hyperosmotic stress also prevented the activation of S6K by both 12-O-tetradecanoylphorbol-13-acetate and anisomycin. Comparison of sorbitol and rapamycin revealed that both agents inactivated S6K and caused dephosphorylation of Ser/Thr-Pro sites in the COOH terminus of S6K, including Thr(412), a residue essential to S6K regulation, as determined by phospho specific antibodies. Rapamycin-resistant S6K truncation mutants were similarly resistant to deactivation by sorbitol. Additionally, the PHAS-1 mobility shift, which is sensitive to rapamycin, was also found to be sensitive to osmotic stress. Experiments using the p38 inhibitor SB203580 and dominant negative mutants involving both stress-activated protein kinase/c-Jun NH(2)-terminal kinase and p38 stress pathways indicated that these pathways are probably not involved in osmotic stress inhibition of S6K. Examining the potential involvement of a phosphatase, we found that sodium pyrophosphate, sodium vanadate, cyclosporin A, tautomycin, and okadaic acid had no effect on osmotic stress inhibition of S6K. However, calyculin A prevented both rapamycin- and sorbitol mediated deactivation of S6K. Our results suggest that osmotic stress and rapamycin act through a calyculin A-sensitive phosphatase to cause dephosphorylation and deactivation of S6K. PMID- 10455143 TI - Interdomain signaling in a two-domain fragment of the human glucocorticoid receptor. AB - Studies of individual domains or subdomains of the proteins making up the nuclear receptor family have stressed their modular nature. Nevertheless, these receptors function as complete proteins. Studies of specific mutations suggest that in the holoreceptors, intramolecular domain-domain interactions are important for complete function, but there is little knowledge concerning these interactions. The important transcriptional transactivation function in the N-terminal part of the glucocorticoid receptor (GR) appears to have little inherent structure. To study its interactions with the DNA binding domain (DBD) of the GR, we have expressed the complete sequence from the N-terminal through the DBD of the human GR. Circular dichroism analyses of this highly purified, multidomain protein show that it has a considerable helical content. We hypothesized that binding of its DBD to the cognate glucocorticoid response element would confer additional structure upon the N-terminal domain. Circular dichroism and fluorescence emission studies suggest that additional helicity as well as tertiary structure occur in the two-domain protein upon DNA binding. In sum, our data suggest that interdomain interactions consequent to DNA binding imparts structure to the portion of the GR that contains a major transactivation domain. PMID- 10455144 TI - Effects of S-nitrosation on oxygen binding by normal and sickle cell hemoglobin. AB - S-Nitrosated hemoglobin (SNO-Hb) is of interest because of the allosteric control of NO delivery from SNO-Hb made possible by the conformational differences between the R- and T-states of Hb. To better understand SNO-Hb, the oxygen binding properties of S-nitrosated forms of normal and sickle cell Hb were investigated. Spectral assays and electrospray ionization mass spectrometry were used to quantify the degree of S-nitrosation. Hb A(0) and unpolymerized Hb S exhibit similar shifts toward their R-state conformations in response to S nitrosation, with increased oxygen affinity and decreased cooperativity. Responses to 2, 3-diphosphoglycerate were unaltered, indicating regional changes in the deoxy structure of SNO-Hb that accommodate NO adduction. A cycle of deoxygenation/reoxygenation does not cause loss of NO or appreciable heme oxidation. There is, however, appreciable loss of NO and heme oxidation when oxygen-binding experiments are carried out in the presence of glutathione. These results indicate that the in vivo stability of SNO-Hb and its associated vasoactivity depend on the abundance of thiols and other factors that influence transnitrosation reactions. The increased oxygen affinity and R-state character that result from S-nitrosation of Hb S would be expected to decrease its polymerization and thereby lessen the associated symptoms of sickle cell disease. PMID- 10455145 TI - Structure of d(CGCGAATTCGCG) in the presence of Ca(2+) ions. AB - The dodecamer d(CGCGAATTCGCG) was the first oligonucleotide to be crystallized as a B-DNA duplex. Its structure was analyzed in detail in the early 1980s. Here we show that, in the presence of Ca(2+), it crystallizes in a different way (R3 space group). The dodecamers form parallel columns of straight duplexes with ten base pairs in the B form. The terminal cytosines in each molecule are disordered, whereas the terminal guanines are placed in the minor groove of neighbor duplexes. The central GAATTC region is practically identical to that found in the classic structure of the same dodecamer crystallized in the P2(1)2(1)2(1) space group in the presence of Mg(2+) and spermine. Its structure is thus independent of the crystallization conditions which have been used. PMID- 10455147 TI - The transmembrane domains of the human multidrug resistance P-glycoprotein are sufficient to mediate drug binding and trafficking to the cell surface. AB - The human multidrug resistance P-glycoprotein (P-gp) is organized in two tandem repeats with each repeat consisting of an N-terminal hydrophobic domain containing six potential transmembrane segments followed by a hydrophilic domain containing a nucleotide-binding fold. A series of deletion mutants together with an in vivo drug-binding assay were used to test whether the deletion mutants interacted with substrates or were transported to the cell surface. We found that a deletion mutant consisting of only the transmembrane domains (residues 1-379 plus 681-1025) retained the ability to interact with drug substrates. In the absence of drug substrates, the deletion mutant was sensitive to trypsin and endoglycosidase H. Expression in the presence of verapamil, vinblastine, capsaicin, or cyclosporin A, however, resulted in a mutant protein that was resistant to trypsin and endoglycosidase H. The mutant was then detected at the cell surface and was sensitive to digestion by endoglycosidase F. By contrast, the N-terminal transmembrane domain (residues 1-379) alone did not interact with drug substrates, since it was sensitive to only endoglycosidase H and was not detected at the cell surface. These results show that the nucleotide-binding domains are not required for interaction of P-gp with substrate or for trafficking of P-gp to the cell surface. PMID- 10455146 TI - cAMP-induced phosphorylation and inhibition of Na(+)/H(+) exchanger 3 (NHE3) are dependent on the presence but not the phosphorylation of NHE regulatory factor. AB - The members of the regulatory factor (RF) gene family, Na(+)/H(+) exchanger (NHE) RF and NHE3 kinase A regulatory factor (E3KARP) are necessary for cAMP to inhibit the epithelial brush border NHE isoform 3 (NHE3). The mechanism of their action was studied using PS120 fibroblasts stably transfected with rabbit NHE3 and wild type rabbit NHE-RF or wild type human E3KARP. 8-Bromo-cAMP (8-Br-cAMP) had no effect on Na(+)/H(+) exchange activity in cells expressing NHE3 alone. In contrast, in cells co-expressing NHE-RF, 8-Br-cAMP inhibited NHE3 by 39%. In vivo phosphorylation of NHE3 demonstrated that cAMP increased phosphorylation in two chymotrypsin-generated phosphopeptides of NHE3 in cells containing NHE-RF or E3KARP but not in cells lacking these proteins. The requirement for phosphorylation of NHE-RF in this cAMP-induced inhibition of NHE3 was examined by studying a mutant NHE-RF in which serines 287, 289, and 290 were mutated to alanines. Wild type NHE-RF was a phosphorylated protein under basal conditions, but treatment with 8-Br-cAMP did not alter its phosphorylation. Mutant NHE-RF was not phosphorylated either under basal conditions or after 8-Br-cAMP. 8-Br-cAMP inhibited NHE3 similarly in PS120/NHE3 cells containing wild type or mutant NHE RF. NHE-RF and NHE3 co-precipitated and did so similarly with and without cAMP. Mutant NHE-RF also similarly immunoprecipitated NHE3 in the presence and absence of 8-Br-cAMP. This study shows that members of the regulatory factor gene family, NHE-RF and E3KARP, are necessary for cAMP inhibition of NHE3 by allowing NHE3 to be phosphorylated. This inhibition is not dependent on the phosphorylation of NHE RF. PMID- 10455148 TI - A novel oncostatin M-inducible gene OIG37 forms a gene family with MyD118 and GADD45 and negatively regulates cell growth. AB - Oncostatin M (OSM) is a member of the IL-6 family cytokines that use gp130 as a common signal transducer and exhibits both growth stimulatory as well as growth inhibitory activity depending on the cells. To analyze the mechanism of OSM function, we isolated immediate early responsive genes upon OSM stimulation. Here we describe the novel OSM-inducible gene OIG37 that is related to MyD118 and GADD45. The MyD118 gene has been described as an immediate early gene induced by IL-6 in M1 monocytic cells, and GADD45 was identified as a gene induced by UV or gamma-ray irradiation. Both are considered to function in growth arrest and/or DNA repair. Although the expression of OIG37, MyD118, and GADD45 was rather ubiquitous, it was differentially regulated. As the gp130 mutant defective for activating the STAT3 pathway showed the reduced induction of OIG37 by cytokine stimulation and expression of dominant negative STAT3 inhibited the induction of OIG37 by OSM, STAT3 is involved in OIG37 induction by IL-6 family cytokines. To examine the function of OIG37, we expressed it in NIH3T3 and IL-3-dependent BaF3 cells and found that OIG37 suppressed cell growth without any evidence of apoptosis. Whereas both MyD118 and OIG37 suppressed cell growth in both cell lines, suppression by OIG37 was more efficient than by MyD118. Immunoprecipitation experiments indicated that OIG37 associates with p21, a cyclin-dependent kinase inhibitor, and proliferating cell nuclear antigen. PMID- 10455149 TI - An erythropoietin fusion protein comprised of identical repeating domains exhibits enhanced biological properties. AB - The hematopoietic growth factor erythropoietin (Epo) initiates its intracellular signaling cascade by binding to and inducing the homodimerization of two identical receptor molecules. We have now constructed and expressed in COS cells a cDNA encoding a fusion protein consisting of two complete human Epo domains linked in tandem by a 17-amino acid flexible peptide. On SDS-polyacrylamide gel electrophoresis, the Epo-Epo fusion protein migrated as a broad band with an average apparent molecular mass of 76 kDa, slightly more than twice the average apparent molecular mass of Epo, 37 kDa. Enzymatic N-deglycosylation resulted in an Epo-Epo species that migrated on SDS-polyacrylamide gel electrophoresis as a narrow band with an average apparent molecular mass of 39 kDa. The specific activity of the Epo-Epo fusion protein in vitro (1,007 IU/microgram; 76 IU/pmol) was significantly greater than that of Epo (352 IU/microgram; 13 IU/pmol). Moreover, secretion of Epo-Epo by COS cells was 8-fold greater than that of Epo. Subcutaneous administration of a single dose of Epo-Epo to mice resulted in a significant increase in red blood cell production within 7 days. In contrast, administration of an equivalent dose of conventional recombinant Epo was without effect. The pharmacokinetic behavior of Epo-Epo differed significantly from that of Epo. The results suggest that Epo-Epo may have important biological and therapeutic advantages. PMID- 10455150 TI - Human transcription release factor 2 dissociates RNA polymerases I and II stalled at a cyclobutane thymine dimer. AB - RNA polymerase II stalled at a lesion in the transcribed strand is thought to constitute a signal for transcription-coupled repair. Transcription factors that act on RNA polymerase in elongation mode potentially influence this mode of repair. Previously, it was shown that transcription elongation factors TFIIS and Cockayne's syndrome complementation group B protein did not disrupt the ternary complex of RNA polymerase II stalled at a thymine cyclobutane dimer, nor did they enable RNA polymerase II to bypass the dimer. Here we investigated the effect of the transcription factor 2 on RNA polymerase II and RNA polymerase I stalled at thymine dimers. Transcription factor 2 is known to release transcripts from RNA polymerase II early elongation complex generated by pulse-transcription. We found that factor 2 (which is also called release factor) disrupts the ternary complex of RNA polymerase II at a thymine dimer and surprisingly exerts the same effect on RNA polymerase I. These findings show that in mammalian cells a RNA polymerase I or RNA polymerase II transcript truncated by a lesion in the template strand may be discarded unless repair is accomplished rapidly by a mechanism that does not displace stalled RNA polymerases. PMID- 10455151 TI - Structural identification of a novel pro-inflammatory epoxyisoprostane phospholipid in mildly oxidized low density lipoprotein. AB - One of the earliest steps in the development of the atherosclerotic lesion is the accumulation of monocyte/macrophages within the vessel wall. Oxidized lipids present in minimally modified-low density lipoproteins (MM-LDL) contribute to this process by activating endothelial cells to express monocyte-specific adhesion molecules and chemoattractant factors. A major focus of our group has been the isolation and characterization of the biologically active oxidized lipids in MM-LDL. We have previously characterized three oxidized phospholipids present in MM-LDL, atherosclerotic lesions of fat fed rabbits, and autoxidized 1 palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (Ox-PAPC) that induced human aortic endothelial cells to adhere human monocytes in vitro. We have used sequential normal and reverse phase-high performance liquid chromatography to isolate various isomers of an oxidized phospholipid from autoxidized 1-palmitoyl 2-arachidonoyl-sn-glycero-3-phosphocholine. The fatty acid in the sn-2 position of this biologically active isomer and its dehydration product was released by phospholipase A(2) and characterized. Hydrogenation with platinum(IV) oxide/hydrogen suggested a cyclic moiety, and reduction with sodium borohydride suggested two reducible oxygen-containing groups in the molecule. The fragmentation pattern produced by electrospray ionization-collision induced dissociation-tandem mass spectrometry was consistent with a molecule resembling an E-ring prostaglandin with an epoxide at the 5,6 position. The structure of this lipid was confirmed by proton nuclear magnetic resonance spectroscopy analysis of the free fatty acid isolated from the dehydration product of m/z 828.5. Based on these studies, we arrived at the structure of the biologically active oxidized phospholipids as 1-palmitoyl-2-(5, 6-epoxyisoprostane E(2))-sn glycero-3-phosphocholine. The identification of this molecule adds epoxyisoprostanes to the growing list of biologically active isoprostanes. PMID- 10455152 TI - Rapid nucleolytic degradation of the small cytoplasmic Y RNAs during apoptosis. AB - We have investigated the fate of the RNA components of small ribonucleoprotein particles in apoptotic cells. We show that the cytoplasmic Ro ribonucleoprotein associated Y RNAs are specifically and rapidly degraded during apoptosis via a caspase-dependent mechanism. This is the first study describing the selective degradation of a specific class of small structural RNA molecules in apoptotic cells. Cleavage and subsequent truncation of Y RNAs was observed upon exposure of cells to a variety of apoptotic stimuli and were found to be inhibited by Bcl-2, zinc, and several caspase inhibitors. These results indicate that apoptotic degradation of Y RNAs is dependent on caspase activation, which suggests that the nucleolytic activity responsible for hY RNA degradation is activated downstream of the caspase cascade. The Y RNA degradation products remain bound by the Ro60 protein and in part also by the La protein, the only two proteins known to be stably associated with intact Ro ribonucleoprotein particles. The size of the Y RNA degradation products is consistent with the protection from degradation of the most highly conserved region of the Y RNAs by the bound Ro60 and La proteins. Our results indicate that the rapid abrogation of the yet unknown function of Y RNAs might be an early step in the systemic deactivation of the dying cell. PMID- 10455153 TI - CCAAT/enhancer-binding proteins are mediators in the protein kinase A-dependent activation of the decidual prolactin promoter. AB - In the course of decidualization, human endometrial stromal cells (ESC) activate the alternative upstream promoter of the decidual prolactin (dPRL) gene. The dPRL promoter is induced by the protein kinase A pathway in a delayed fashion via the region -332/-270 which contains two overlapping consensus binding sequences, B and D, for CCAAT/enhancer-binding proteins (C/EBP). Here we show that sites B and D both bind C/EBPbeta and -delta from ESC nuclear extracts. When decidualization of cultured ESC was induced by treatment with 8-Br-cAMP, complex formation on sites B and D was enhanced. Western blot analysis revealed an elevation of both C/EBPbeta isoforms, liver-enriched activator protein and liver-enriched inhibitory protein, with a delayed onset between 8 and 24 h of cAMP treatment, while C/EBPdelta expression remained unaffected. Cyclic AMP-mediated activation of dPRL promoter construct dPRL-332/luc3 was abrogated by mutation of sites B and D at -310/-285. An expression vector for liver-enriched activator protein potently induced transcription of dPRL-332/luc3 and further enhanced cAMP mediated induction, while liver-enriched inhibitory protein expression vector abolished the cAMP response, implying that C/EBPs serve as mediators in the delayed cAMP signal transduction to the dPRL promoter. The ratio between activating and repressing isoforms is likely to dictate the transcriptional output. PMID- 10455154 TI - Human papillomavirus type 16 E6-enhanced susceptibility of L929 cells to tumor necrosis factor alpha correlates with increased accumulation of reactive oxygen species. AB - Human papillomavirus type 16 (HPV-16) E6 has been shown to prevent or enhance apoptosis depending on the stimulus and cell type. Here we present evidence that HPV-16 E6 sensitized murine fibrosarcoma L929 cells to tumor necrosis factor alpha (TNF)-induced cytolysis. The E6-enhanced cytolysis correlated with a precedent increase in reactive oxygen species (ROS) level and antioxidant treatment could completely block the E6-dependent sensitization. These findings represent the first demonstration of a link between a viral oncogene-sensitized cytolysis and ROS. Previous studies have shown conflicting results regarding whether TNF-induced cytolysis of L929 cells is through necrosis or apoptosis. Here we report that, although L929 cells underwent DNA fragmentation after exposure to TNF, they retained the morphology of intact nuclei while gaining permeability to propidium iodide, features characteristic of necrosis rather than apoptosis. We confirmed that the broad spectrum caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone markedly increased the susceptibility of L929 cells to TNF, and further demonstrated that E6 enhanced this susceptibility, which again correlated with increased ROS accumulation. We showed that the expression of E6 in L929 cells did not alter the stability of p53, and the cells retained a p53 response to actinomycin D. Furthermore, two E6 mutants defective for p53 degradation in other systems exhibited differential effects on TNF sensitization. These results suggest that the enhancement of TNF induced L929 cytolysis by E6 is independent of p53 degradation. We also found that TNF-induced activation of NF-kappaB did not account for the enhanced TNF susceptibility by E6. PMID- 10455155 TI - Presence of phospholipid-neutral lipid complex structures in atherosclerotic lesions as detected by a novel monoclonal antibody. AB - A novel monoclonal antibody (ASH1a/256C) that recognizes atherosclerotic lesions in human and Watanabe heritable hyperlipidemic (WHHL) rabbit aortae is described. When (123)I-labeled ASH1a/256C antibody is injected intravenously into WHHL rabbits, it associates specifically with fatty streaks on the aorta. The antigen recognized by the antibody is lipid, based on extraction with chloroform and methanol from WHHL rabbit tissues. The antigen, purified by high performance liquid chromatography, was shown to be phosphatidylcholine (PC), which contains unsaturated fatty acyl groups based on analyses utilizing (1)H and (13)C nuclear magnetic resonance, Fourier transfer-infrared spectrum, and mass spectrometry. The antibody did not react with other classes of phospholipids or neutral lipids when tested using an enzyme-linked immunosorbent assay. When PC was mixed with either cholesterol, cholesteryl ester, or triacylglycerol, however, the reactivity of the antibody to PC increased up to 8-fold. Homogenates of aorta tissue obtained from normal and WHHL rabbits were fractionated using sucrose density gradient ultracentrifugation in which neutral lipid droplets, cellular membranes, and proteins are separated. The phospholipid content in cellular membrane fractions from WHHL rabbits was twice as high as that of normal rabbits, and there was an enormous difference in the antigenic activity in these fractions. The content of cholesterol in the cellular membrane fraction of WHHL rabbits was approximately 50 times higher than that of normal rabbits. Addition of neutral lipids to the cellular membrane fraction of normal rabbit markedly increased the antigenic activity. Atheromatous lesions in thickened WHHL rabbit aortic intima that were rich in lipid droplets were stained positively with ASH1a/256C immunohistochemically. These results strongly suggest that PC-neutral lipid complex domains are formed in atherosclerotic lesions. PMID- 10455156 TI - A novel glycosulfopeptide binds to P-selectin and inhibits leukocyte adhesion to P-selectin. AB - P-selectin glycoprotein ligand-1 (PSGL-1) is a dimeric membrane mucin on leukocytes that binds selectins. The molecular features of PSGL-1 that determine this high affinity binding are unclear. Here we demonstrate the in vitro synthesis of a novel glycosulfopeptide (GSP-6) modeled after the extreme N terminus of PSGL-1, which has been predicted to be important for P-selectin binding. GSP-6 contains three tyrosine sulfate (TyrSO(3)) residues and a monosialylated, core 2-based O-glycan with a sialyl Lewis x (C2-O-sLe(x)) motif at a specific Thr residue. GSP-6 binds tightly to immobilized P-selectin, whereas glycopeptides lacking either TyrSO(3) or C2-O-sLe(x) do not detectably bind. Remarkably, an isomeric glycosulfopeptide to GSP-6, termed GSP-6', which contains sLe(x) on an extended core 1-based O-glycan, does not bind immobilized P selectin. Equilibrium gel filtration analysis revealed that GSP-6 binds to soluble P-selectin with a K(d) of approximately 350 nM. GSP-6 (<5 microM) substantially inhibits neutrophil adhesion to P-selectin in vitro, whereas free sLe(x) (5 mM) only slightly inhibits adhesion. In contrast to the inherent heterogeneity of post-translational modifications of recombinant proteins, glycosulfopeptides permit the placement of sulfate groups and glycans of precise structure at defined positions on a polypeptide. This approach should expedite the probing of structure-function relationships in sulfated and glycosylated proteins, and may facilitate development of novel drugs to treat inflammatory diseases involving P-selectin-mediated leukocyte adhesion. PMID- 10455157 TI - A set of highly conserved RNA-binding proteins, alphaCP-1 and alphaCP-2, implicated in mRNA stabilization, are coexpressed from an intronless gene and its intron-containing paralog. AB - Gene families normally expand by segmental genomic duplication and subsequent sequence divergence. Although copies of partially or fully processed mRNA transcripts are occasionally retrotransposed into the genome, they are usually nonfunctional ("processed pseudogenes"). The two major cytoplasmic poly(C) binding proteins in mammalian cells, alphaCP-1 and alphaCP-2, are implicated in a spectrum of post-transcriptional controls. These proteins are highly similar in structure and are encoded by closely related mRNAs. Based on this close relationship, we were surprised to find that one of these proteins, alphaCP-2, was encoded by a multiexon gene, whereas the second gene, alphaCP-1, was identical to and colinear with its mRNA. The alphaCP-1 and alphaCP-2 genes were shown to be single copy and were mapped to separate chromosomes. The linkage groups encompassing each of the two loci were concordant between mice and humans. These data suggested that the alphaCP-1 gene was generated by retrotransposition of a fully processed alphaCP-2 mRNA and that this event occurred well before the mammalian radiation. The stringent structural conservation of alphaCP-1 and its ubiquitous tissue distribution suggested that the retrotransposed alphaCP-1 gene was rapidly recruited to a function critical to the cell and distinct from that of its alphaCP-2 progenitor. PMID- 10455158 TI - Extracellular signal-regulated protein kinase activation is required for the anti hypertrophic effect of atrial natriuretic factor in neonatal rat ventricular myocytes. AB - Atrial natriuretic factor (ANF) inhibits proliferation in non-myocardial cells and is thought to be anti-hypertrophic in cardiomyocytes. We investigated the possibility that the anti-hypertrophic actions of ANF involved the mitogen activated protein kinase signal transduction cascade. Cultured neonatal rat ventricular myocytes treated for 48 h with the alpha(1)-adrenergic agonist phenylephrine (PE) had an 80% increase in cross-sectional area (CSA). ANF alone had no effect but inhibited PE-induced increases in CSA by approximately 50%. The mitogen-activated protein kinase/ERK kinase (MEK) inhibitor PD098059 minimally inhibited PE-induced increases in CSA, but it completely abolished ANF-induced inhibition of PE-induced increases. ANF-induced extracellular signal-regulated protein kinase (ERK) nuclear translocation was also eliminated by PD098059. ANF treatment caused MEK phosphorylation and activation but failed to activate any of the Raf isoforms. ANF induced a rapid increase in ERK phosphorylation and in vitro kinase activity. PE also increased ERK activity, and the combined effect of ANF and PE appeared to be additive. ANF-induced ERK phosphorylation was eliminated by PD098059. ANF induced minimal phosphorylation of JNK or p38, indicating that its effect on ERK was specific. ANF-induced activation of ERK was mimicked by cGMP analogs, suggesting that ANF-induced ERK activation involves the guanylyl cyclase activity of the ANF receptor. These data suggest that there is an important linkage between cGMP signaling and the mitogen-activated protein kinase cascade and that selective ANF activation of ERK is required for the anti hypertrophic action of ANF. Thus, ANF expression might function as the natural defense of the heart against maladaptive hypertrophy through its ability to activate ERK. PMID- 10455159 TI - Nuclear localization of protein kinase U-alpha is regulated by 14-3-3. AB - 14-3-3 proteins are intracellular, dimeric molecules that bind to and modify the activity of several signaling proteins. We used human 14-3-3zeta as a bait in the yeast two-hybrid system to screen a murine embryonic cDNA library. One interacting clone was found to encode the carboxyl terminus of a putative protein kinase. The coding sequence of the human form (protein kinase Ualpha, PKUalpha) of this protein kinase was found in GenBank(TM) on the basis of sequence homology. The two-hybrid clone was also highly homologous to TOUSLED, an Arabidopsis thaliana protein kinase that is required for normal flower and leaf development. PKUalpha has been found by coimmunoprecipitation to bind to 14-3 3zeta in vivo. Our confocal laser immunofluorescence microscopic experiments revealed that PKUalpha colocalizes with the cytoplasmic intermediate filament system of cultured fibroblasts in the G(1) phase of the cell cycle. PKUalpha is found in the perinuclear area of S phase cells and in the nucleus of late G(2) cells. Transfection of cells with a dominant negative form of 14-3-3eta promotes the nuclear localization of PKUalpha. These results suggest that the subcellular localization of PKUalpha is regulated, at least in part, by its association with 14-3-3. PMID- 10455160 TI - Truncation of the C terminus of the rat brain Na(+)-Ca(2+) exchanger RBE-1 (NCX1.4) impairs surface expression of the protein. AB - The C terminus of the rat brain Na(+)-Ca(2+) exchanger (RBE-1; NCX1. 4) (amino acids 875-903) is modeled to contain the last transmembrane alpha helix (amino acids 875-894) and an intracellular extramembraneous tail of 9 amino acids (895 903). Truncation of the last 9 C-terminal amino acids, Glu-895 to stop, did not significantly impair functional expression in HeLa or HEK 293 cells. Truncation, however, of 10 amino acids (Leu-894 to stop; mutant C10) reduced Na(+) gradient dependent Ca(2+) uptake to 35-39% relative to the wild type parent exchanger, and further truncation of 13 or more amino acids resulted in expression of trace amounts of transport activity. Western analysis indicated that Na(+)-Ca(2+) exchanger protein was produced whether transfection was carried out with functional or non-functional mutants. Immunofluorescence studies of HEK 293 cells expressing N-Flag epitope-tagged wild type and mutant Na(+)-Ca(2+) exchangers revealed that transport activity in whole cells correlated with surface expression. All cells expressing the wild type exchanger or C9 exhibited surface expression of the protein. Only 39% of the cells expressing C10 exhibited surface expression, and none was detected in cells transfected with non-functional mutants C13 and C29. Since functional and non-functional mutants were glycosylated, the C terminus is not mandatory to translocation into the endoplasmic reticulum (ER). Endoglycosidase H digestion of [(35)S]methionine labeled protein derived from wild type Na(+)-Ca(2+) exchanger and from C10 indicated that resistance to the digestion was acquired after 1 and 5 h of chase, respectively. C29 did not acquire detectable resistance to endoglycosidase H digestion even after 10 h of chase. Taken together, these results suggest that the "cellular quality control machinery" can tolerate the structural change introduced by truncation of the C terminus up to Ser-893 albeit with reduced rate of ER-->Golgi transfer and reduced surface expression of the truncated protein. Further truncation of C-terminal amino acids leads to retention of the truncated protein in the ER, no transfer to the Golgi, and no surface expression. PMID- 10455161 TI - Effect of species differences on stromelysin-1 (MMP-3) inhibitor potency. An explanation of inhibitor selectivity using homology modeling and chimeric proteins. AB - For an animal model to predict a compound's potential for treating human disease, inhibitor interactions with the cognate enzymes of separate species must be comparable. Rabbit and human isoforms of stromelysin-1 are highly homologous, yet there are clear and significant compound-specific differences in inhibitor potencies between these two enzymes. Using crystal structures of discordant inhibitors complexed with the human enzyme, we generated a rabbit enzyme homology model that was used to identify two unmatched residues near the active site that could explain the observed disparities. To test these observations, we designed and synthesized three chimeric mutants of the human enzyme containing the single (H224N and L226F) and double (H224N/L226F) mutations. A comparison of inhibitor potencies among the mutant and wild-type enzymes shows that the mutation of a single amino acid in the human enzyme, histidine 224 to asparagine, is sufficient to change the selectivity profile of the mutant to that of the rabbit isoform. These studies emphasize the importance of considering species differences, which can result from even minor protein sequence variations, for the critical enzymes in an animal disease model. Homology modeling provides a tool to identify key differences in isoforms that can significantly affect native enzyme activity. PMID- 10455162 TI - The protozoan parasite Toxoplasma gondii expresses two functional plant-like glycolytic enzymes. Implications for evolutionary origin of apicomplexans. AB - The recent discovery of a vestigial, nonphotosynthetic plastid ("apicoplast") in the Apicomplexa has considerably modified our perception of the evolutionary origin of these parasites. Phylogenetic analysis and the presence of four surrounding membranes of the apicoplast provide important support for the hypothesis that apicomplexans have acquired their apicoplast by secondary endosymbiosis, probably from a green alga. This suggests that genes encoding predicted homologs of proteins of green algae or related photosynthetic lineages could have entered the nucleus of apicomplexan parasites by transfer from the ancestor harboring the apicoplast. We describe here complementary DNAs encoding two Toxoplasma gondii glycolytic enzymes, glucose-6-phosphate isomerase (G6-PI) and enolase, which have considerable identities with land plant counterparts. Both cDNAs of T. gondii complement Escherichia coli mutants lacking G6-PI and enolase genes and lead to the expression of active enzymes. In the drug untreatable encysted bradyzoites of T. gondii, G6-PI and enolase genes are overexpressed or exclusively expressed at both transcriptional and protein levels. Moreover, three-dimensional models and protein phylogeny confirmed that G6-PIs and enolases of T. gondii, Plasmodium falciparum, and land plants are closely related. Because these glycolytic enzymes are plant homologs, which differ from those of animals, they will be useful to trace the evolutionary origin of Apicomplexa and might offer novel chemotherapeutic targets in diseases caused by apicomplexan parasites. PMID- 10455163 TI - Insulin and exercise decrease glycogen synthase kinase-3 activity by different mechanisms in rat skeletal muscle. AB - Glycogen synthase activity is increased in response to insulin and exercise in skeletal muscle. Part of the mechanism by which insulin stimulates glycogen synthesis may involve phosphorylation and activation of Akt, serine phosphorylation and deactivation of glycogen synthase kinase-3 (GSK-3), leading to dephosphorylation and activation of glycogen synthase. To study Akt and GSK-3 regulation in muscle, time course experiments on the effects of insulin injection and treadmill running exercise were performed in hindlimb skeletal muscle from male rats. Both insulin and exercise increased glycogen synthase activity (%I form) by 2-3-fold over basal. Insulin stimulation significantly increased Akt phosphorylation and activity, whereas exercise had no effect. The time course of the insulin-stimulated increase in Akt was closely matched by GSK-3alpha Ser(21) phosphorylation and a 40-60% decrease in GSK-3alpha and GSK-3beta activity. Exercise also deactivated GSK-3alpha and beta activity by 40-60%. However, in contrast to the effects of insulin, there was no change in Ser(21) phosphorylation in response to exercise. Tyrosine dephosphorylation of GSK-3, another putative mechanism for GSK-3 deactivation, did not occur with insulin or exercise. These data suggest the following: 1) GSK-3 is constitutively active and tyrosine phosphorylated under basal conditions in skeletal muscle, 2) both exercise and insulin are effective regulators of GSK-3 activity in vivo, 3) the insulin-induced deactivation of GSK-3 occurs in response to increased Akt activity and GSK-3 serine phosphorylation, and 4) there is an Akt-independent mechanism for deactivation of GSK-3 in skeletal muscle. PMID- 10455164 TI - Novel mechanism of inhibition of elastase by beta-lactams is defined by two inhibitor crystal complexes. AB - Two structurally related beta-lactams form different covalent complexes upon reaction with porcine elastase. The high resolution x-ray structures of these two complexes provide a clear insight into the mechanism of the reaction and suggest the design of a new class of serine protease inhibitors that resist enzyme reactivation by hydrolysis of the acyl intermediate. The presence of a hydroxyethyl substituent on the beta-lactam ring provides a new reaction pathway resulting in the elimination of the hydroxyethyl group and the formation of a stabilizing conjugated double bond system. In contrast, the presence of a diethyl substituent on the beta-lactam ring leads to addition of water. The two enzyme complexes show very different binding modes in the enzyme active site. PMID- 10455165 TI - Trench-shaped binding sites promote multiple classes of interactions between collagen and the adherence receptors, alpha(1)beta(1) integrin and Staphylococcus aureus cna MSCRAMM. AB - Most mammalian cells and some pathogenic bacteria are capable of adhering to collagenous substrates in processes mediated by specific cell surface adherence molecules. Crystal structures of collagen-binding regions of the human integrin alpha(2)beta(1) and a Staphylococcus aureus adhesin reveal a "trench" on the surface of both of these proteins. This trench can accommodate a collagen triple helical structure and presumably represents the ligand-binding site (Emsley, J., King, S. L., Bergelson, J. M., and Liddington, R. C. (1997) J. Biol. Chem. 272, 28512-28517; Symersky, J., Patti, J. M., Carson, M., House-Pompeo, K., Teale, M., Moore, D., Jin, L., Schneider, A., DeLucas, L. J., Hook, M., and Narayana, S. V. L. (1997) Nat. Struct. Biol. 4, 833-838). We report here the crystal structure of the alpha subunit I domain from the alpha(1)beta(1) integrin. This collagen binding protein also contains a trench on one face in which the collagen triple helix may be docked. Furthermore, we compare the collagen-binding mechanisms of the human alpha(1) integrin I domain and the A domain from the S. aureus collagen adhesin, Cna. Although the S. aureus and human proteins have unrelated amino acid sequences, secondary structure composition, and cation requirements for effective ligand binding, both proteins bind at multiple sites within one collagen molecule, with the sites in collagen varying in their affinity for the adherence molecule. We propose that (i) these evolutionarily dissimilar adherence proteins recognize collagen via similar mechanisms, (ii) the multisite, multiclass protein/ligand interactions observed in these two systems result from a binding site trench, and (iii) this unusual binding mechanism may be thematic for proteins binding extended, rigid ligands that contain repeating structural motifs. PMID- 10455166 TI - Increased Ser-10 phosphorylation of histone H3 in mitogen-stimulated and oncogene transformed mouse fibroblasts. AB - When the Ras mitogen-activated protein kinase (MAPK) signaling pathway of quiescent cells is stimulated with growth factors or phorbol esters, the early response genes c-fos and c-myc are rapidly induced, and concurrently there is a rapid phosphorylation of histone H3. Using an antibody specific for phosphorylated Ser-10 of H3, we show that Ser-10 of H3 is phosphorylated, and we provide direct evidence that phosphorylated H3 is associated with c-fos and c-myc genes in stimulated cells. H3 phosphorylation may contribute to proto-oncogene induction by modulating chromatin structure and releasing blocks in elongation. Previously we reported that persistent stimulation of the Ras-MAPK signaling pathway in oncogene-transformed cells resulted in increased amounts of phosphorylated histone H1. Here we show that phosphorylated H3 is elevated in the oncogene-transformed mouse fibroblasts. Further we show that induction of ras expression results in a rapid increase in H3 phosphorylation. H3 phosphatase, identified as PP1, activities in ras-transformed and parental fibroblast cells were similar, suggesting that elevated H3 kinase activity was responsible for the increased level of phosphorylated H3 in the oncogene-transformed cells. Elevated levels of phosphorylated H1 and H3 may be responsible for the less condensed chromatin structure and aberrant gene expression observed in the oncogene transformed cells. PMID- 10455167 TI - Tetrahydrobiopterin inhibits monomerization and is consumed during catalysis in neuronal NO synthase. AB - The biosynthesis of nitric oxide (NO) is catalyzed by homodimeric NO synthases (NOS). For unknown reasons, all NOS co-purify with substoichiometric amounts of (6R)-5,6,7,8-tetrahydrobiopterin (H(4)Bip) and require additional H(4)Bip for maximal activity. We examined the effects of H(4)Bip and pterin-derived inhibitors (anti-pterins) on purified neuronal NOS-I quaternary structure and H(4)Bip content. During L-arginine turnover, NOS-I dimers time dependently dissociated into inactive monomers, paralleled by a loss of enzyme-associated pterin. Dimer dissociation was inhibited when saturating levels of H(4)Bip were added during catalysis. Similar results were obtained with pterin-free NOS-I expressed in Escherichia coli. This stabilizing effect of H(4)Bip was mimicked by the anti-pterin 2-amino-4,6-dioxo-3,4,5,6,8,8a,9, 10-octahydro-oxazolo[1,2f] pteridine (PHS-32), which also displaced NOS-associated H(4)Bip in a competitive manner. Surprisingly, H(4)Bip not only dissociated from NOS during catalysis, but was only partially recovered in the solute (50.0 +/- 16.5% of control at 20 min). NOS-associated H(4)Bip appeared to react with a NOS catalysis product to a derivative distinct from dihydrobiopterin or biopterin. Under identical conditions, reagent H(4)Bip was chemically stable and fully recovered (95.5 +/- 3.4% of control). A similar loss of both reagent and enzyme-bound H(4)Bip and dimer content was observed by NO generated from spermine NONOate. In conclusion, we propose a role for H(4)Bip as a dimer-stabilizing factor of neuronal NOS during catalysis, possibly by interfering with enzyme destabilizing products. PMID- 10455168 TI - Activated neutrophils induce hyperpermeability and phosphorylation of adherens junction proteins in coronary venular endothelial cells. AB - The endothelial adherens junction is formed by complexes of transmembrane adhesive proteins, of which beta-catenin is known to connect the junctional protein vascular endothelial (VE)-cadherin to the cytoskeleton and to play a signaling role in the regulation of junction-cytoskeleton interaction. In this study, we investigated the effect of neutrophil activation on endothelial monolayer integrity and on beta-catenin and VE-cadherin modification. Treatment of cultured bovine coronary endothelial monolayers with C5a-activated neutrophils resulted in an increase in permeability as measured by albumin clearance across the monolayer. Furthermore, large scale intercellular gap formation was observed in coincidence with the hyperpermeability response. Immunofluorescence analysis showed that beta-catenin and VE-cadherin staining changed from a uniform distribution along the membrane of control cells to a diffuse pattern for both proteins and finger-like projections for beta-catenin in neutrophil-exposed monolayers. Correlatively, there was an increase in actin stress fiber formation in treated cells. Finally, beta-catenin and VE-cadherin from neutrophil-treated endothelial cells showed a significant increase in tyrosine phosphorylation. Our results are the first to link neutrophil-mediated changes in adherens junctions with intercellular gap formation and hyperpermeability in microvascular endothelial cells. These data suggest that neutrophils may regulate endothelial barrier function through a process conferring conformational changes to beta catenin and VE-cadherin. PMID- 10455169 TI - Stimulation of mitogen-activated protein kinase by G protein-coupled alpha(2) adrenergic receptors does not require agonist-elicited endocytosis. AB - Agonist-elicited receptor sequestration is strikingly different for the alpha(2A) versus alpha(2B)-adrenergic receptor (alpha(2)-AR) subtypes; the alpha(2B)-AR undergoes rapid and extensive disappearance from the HEK 293 cell surface, whereas the alpha(2A)-AR does not (Daunt, D. A., Hurt, C., Hein, L., Kallio, J., Feng, F., and Kobilka, B. K. (1997) Mol. Pharmacol. 51, 711-720; Eason, M. G., and Liggett, S. B. (1992) J. Biol. Chem. 267, 25473-25479). Since recent reports suggest that endocytosis is required for some G protein-coupled receptors to stimulate the mitogen-activated protein (MAP) kinase cascade (Daaka, Y., Luttrell, L. M., Ahn, S., Della Rocca, G. J., Ferguson, S. S., Caron, M. G., and Lefkowitz, R. J. (1998) J. Biol. Chem. 273, 685-688; Luttrell, L. M., Daaka, Y., Della Rocca, G. J., and Lefkowitz, R. J. (1997) J. Biol. Chem. 272, 31648-31656; Ignatova, E. G., Belcheva, M. M., Bohn, L. M., Neuman, M. C., and Coscia, C. J. (1999) J. Neurosci. 19, 56-63), we evaluated the differential ability of these two subtypes to activate MAP kinase. We observed no correlation between subtype dependent agonist-elicited receptor redistribution and receptor activation of the MAP kinase cascade. Furthermore, incubation of cells with K(+)-depleted medium eliminated alpha(2B)-AR internalization but did not eliminate MAP kinase activation, suggesting that receptor internalization is not a general prerequisite for activation of the MAP kinase cascade via G(i)-coupled receptors. We also noted that neither dominant negative dynamin (K44A) nor concanavalin A treatment dramatically altered MAP kinase activation or receptor redistribution, indicating that these experimental tools do not universally block G protein coupled receptor internalization. PMID- 10455170 TI - A new potent HIV-1 reverse transcriptase inhibitor. A synthetic peptide derived from the interface subunit domains. AB - The biologically relevant and active forms of human immunodeficiency viruses type 1 and 2 reverse transcriptase found in infectious virions are heterodimers produced in a two-step dimerization process. Dimerization involves first the rapid association of the two subunits, followed by a slow conformational change yielding a fully active form. We have shown that the dimeric nature of reverse transcriptase represents a important target for the design of a new class of antiviral agents. In this work, we propose a new strategy for its inhibition by targeting protein/protein interactions during viral formation in infected cells. From the screening of peptides derived from the tryptophan cluster at the interface of the connection subdomain, we have designed a short peptide (10 residues) corresponding to residues 395-404, which can block dimerization of reverse transcriptase in vitro and in infected cells. This peptide is highly efficient in abolishing the production of viral particles, without any adverse toxic side effects, when transduced into human immunodeficiency virus type 1 infected cells together with a new peptide carrier. PMID- 10455171 TI - A novel protease-docking function of integrin at invadopodia. AB - Invadopodia are membrane extensions of aggressive tumor cells that function in the activation of membrane-bound proteases occurring during tumor cell invasion. We explore a novel and provocative activity of integrins in docking proteases to sites of invasion, termed invadopodia. In the absence of collagen, alpha(3)beta(1) integrin and the gelatinolytic enzyme, seprase, exist as nonassociating membrane proteins. Type I collagen substratum induces the association of alpha(3)beta(1) integrin with seprase as a complex on invadopodia. The results show that alpha(3)beta(1) integrin is a docking protein for seprase to form functional invadopodia. In addition, alpha(5)beta(1) integrin may participate in the adhesion process necessary for invadopodial formation. Thus, alpha(3)beta(1) and alpha(5)beta(1) integrins play major organizational roles in the adhesion and formation of invadopodia, promoting invasive cell behavior. PMID- 10455172 TI - Site-directed mutagenesis of the calcium-binding site of blood coagulation factor XIIIa. AB - Blood coagulation factor XIIIa is a calcium-dependent enzyme that covalently ligates fibrin molecules during blood coagulation. X-ray crystallography studies identified a major calcium-binding site involving Asp(438), Ala(457), Glu(485), and Glu(490). We mutated two glutamic acid residues (Glu(485) and Glu(490)) and three aspartic acid residues (Asp(472), Asp(476), and Asp(479)) that are in close proximity. Alanine substitution mutants of these residues were constructed, expressed, and purified from Escherichia coli. The K(act) values for calcium ions increased by 3-, 8-, and 21-fold for E485A, E490A, and E485A,E490A, respectively. In addition, susceptibility to proteolysis was increased by 4-, 9-, and 10-fold for E485A, E490A, and E485A,E490A, respectively. Aspartic acids 472, 476, and 479 are not involved directly in calcium binding since the K(act) values were not changed by mutagenesis. However, Asp(476) and Asp(479) are involved in regulating the conformation for exposure of the secondary thrombin cleavage site. This study provides biochemical evidence that Glu(485) and Glu(490) are Ca(2+)-binding ligands that regulate catalysis. The binding of calcium ion to this site protects the molecule from proteolysis. Furthermore, Asp(476) and Asp(479) play a role in modulating calcium-dependent conformational changes that cause factor XIIIa to switch from a protease-sensitive to a protease-resistant molecule. PMID- 10455173 TI - Purification, characterization, and amino acid sequence of cerato-platanin, a new phytotoxic protein from Ceratocystis fimbriata f. sp. platani. AB - A new phytotoxic protein (cerato-platanin) of about 12.4 kDa has been identified in culture filtrates of the Ascomycete Ceratocystis fimbriata f. sp. platani, the causal agent of canker stain disease. The toxicity of the pure protein was bioassayed by detecting the inducing necrosis in tobacco leaves. The pure protein also elicited host synthesis of fluorescent substances in tobacco and plane (Platanus acerifolia) leaves. We purified the protein from culture medium to homogeneity. Its complete amino acid sequence was determined; this protein consists of 120 amino acid residues, contains 4 cysteines (S-S-bridged), and has a high percentage of hydrophobic residues. The molecular weight calculated from the amino acid sequence agrees with that determined by mass spectrometry, suggesting that no post-transnational modification occurs. Searches performed by the BLAST program in data banks (Swiss-Prot, EBI, and GenBank(TM)) revealed that this protein is highly homologous with two proteins produced by other Ascomycete fungi. One, produced during infection of wheat leaves, is codified by the snodprot1 gene of Phaeosphaeria nodorum (the causal agent of glume blotch of wheat), whereas the other is the rAsp f13 allergen from Aspergillus fumigatus. Furthermore, the N terminus of cerato-platanin is homologous with that of cerato ulmin, a phytotoxic protein belonging to the hydrophobin family and produced by Ophiostoma (Ceratocystis) ulmi, a fungus responsible for Dutch elm disease. PMID- 10455175 TI - Cloning and characterization of novel mouse and human secretory phospholipase A(2)s. AB - Mammalian secretory phospholipase A(2)s (sPLA(2)s) are classified into several groups according to molecular structure and the localization of intramolecular disulfide bridges. Among them, group IIA sPLA(2) has been thought to be one of the key enzymes in the pathogenesis of inflammatory diseases owing to its augmented expression under various inflammatory conditions. However, in a number of inbred mouse strains, the group IIA sPLA(2) gene is naturally disrupted by a frameshift mutation. Here, we report the cloning of a cDNA encoding a novel sPLA(2) expressed in the spleen of group IIA sPLA(2)-deficient mouse. We also cloned its human homolog and mapped its gene location on chromosome 1p36.12 near the loci of group IIA and V sPLA(2) genes. The human mature sPLA(2) protein consists of 125 amino acids (M(r) = 14,500) preceded by a 20-residue prepeptide and is most similar to group IIA sPLA(2) with respect to the number and positions of cysteine residues as well as overall identity (48%). Based on these structural properties, the novel sPLA(2) should be categorized into group II, called group IID to follow the already identified IIA to IIC sPLA(2)s. When the cDNA was expressed in COS-7 cells, PLA(2) activity preferentially accumulated in the culture medium. It is maximally active at neutral to alkaline pH and with 2 mM Ca(2+). In assays with individual substrates, L-alpha-1-palmitoyl-2-linoleoyl phosphatidylethanolamine was more efficiently hydrolyzed than the other phospholipids examined. An RNA blot hybridized with the cDNA exhibited two transcripts (2.0 and 1.0 kb) in human spleen, thymus, and colon. The expression of a novel sPLA(2) mRNA was elevated in the thymus after treatment with endotoxin in rats as well as in group IIA sPLA(2)-deficient mice, suggesting its functional role in the progression of the inflammatory process. PMID- 10455174 TI - Leukemia inhibitory factor and its receptor promote adipocyte differentiation via the mitogen-activated protein kinase cascade. AB - Extracellular factors and intracellular signaling pathways involved in early events of adipocyte differentiation are poorly defined. It is shown herein that expression of leukemia inhibitory factor (LIF) and LIF receptor is developmentally regulated during adipocyte differentiation. Preadipocytes secrete bioactive LIF, and an antagonist of LIF receptor inhibits adipogenesis. Genetically modified embryonic stem (ES) cells combined with culture conditions to commit stem cells into the adipocyte lineage were used to examine the requirement of LIF receptor during in vitro development of adipose cells. The capacity of embryoid bodies derived from lifr(-/-) ES cells to undergo adipocyte differentiation is dramatically reduced. LIF addition stimulates adipocyte differentiation of Ob1771 and 3T3-F442A preadipocytes and that of peroxisome proliferator-activated receptor gamma2 ligand-treated mouse embryonic fibroblasts. Expression of the early adipogenic transcription factors C/EBPbeta and C/EBPdelta is rapidly stimulated following exposure of preadipose cells to LIF. The selective inhibitors of mitogen-activated protein kinase kinase, i.e. PD98059 and U0126, inhibit LIF-induced C/EBP gene expression and prevent adipocyte differentiation induced by LIF. These results are in favor of a model that implicates stimulation of LIF receptor in the commitment of preadipocytes to undergo terminal differentiation by controlling the early expression of C/EBPbeta and C/EBPdelta genes via the mitogen-activated protein kinase cascade. PMID- 10455176 TI - Tyrosine phosphorylation of tub and its association with Src homology 2 domain containing proteins implicate tub in intracellular signaling by insulin. AB - A mutation in the tub gene leads to maturity-onset obesity, insulin resistance, and progressive retinal and cochlear degeneration in mice. tub is a member of a growing family of genes that encode proteins of unknown function that are remarkably conserved across species. The absence of obvious transmembrane domain(s) or signal sequence peptide motif(s) suggests that Tub is an intracellular protein. Additional sequence analysis revealed the presence of putative tyrosine phosphorylation motifs and Src homology 2 (SH2)-binding sites. Here we demonstrate that in CHO-IR cells, transfected Tub is phosphorylated on tyrosine in response to insulin and insulin-like growth factor-1 and that in PC12 cells, insulin but not EGF induced tyrosine phosphorylation of endogenous Tub. In vitro, Tub is phosphorylated by purified insulin receptor kinase as well as by Abl and JAK 2 but not by epidermal growth factor receptor and Src kinases. Furthermore, upon tyrosine phosphorylation, Tub associated selectively with the SH2 domains of Abl, Lck, and the C-terminal SH2 domain of phospholipase Cgamma and insulin enhanced the association of Tub with endogenous phospholipase Cgamma in CHO-IR cells. These data suggest that Tub may function as an adaptor protein linking the insulin receptor, and possibly other protein-tyrosine kinases, to SH2 containing proteins. PMID- 10455177 TI - The carboxyl terminal extension of the Drosophila insulin receptor homologue binds IRS-1 and influences cell survival. AB - The Drosophila insulin receptor (INR) homolog includes an extension of approximately 400 amino acids at the carboxyl-terminal end of its beta subunit containing several tyrosine-based motifs known to mediate interactions with signaling proteins. In order to explore the role of this extension in INR function, mammalian expression vectors encoding either the complete INR beta subunit (beta-Myc) or the INR beta subunit without the carboxyl-terminal extension (betaDelta) were constructed, and the membrane-bound beta subunits were expressed in 293 and Madin-Darby canine kidney cells in the absence of the ligand binding alpha subunits. beta-Myc and betaDelta proteins were constitutively active tyrosine kinases of 180 and 102 kDa, respectively. INR beta-Myc co immunoprecipitated a phosphoprotein of 170 kDa identified as insulin receptor substrate-1 (IRS-1), whereas INR betaDelta did not, suggesting that the site of interaction was within the carboxyl-terminal extension. IRS-1 was phosphorylated on tyrosine to a much greater extent in cells expressing INR beta-Myc than in parental or INR betaDelta cells. Despite this, a variety of PTB or SH2 domain containing signaling proteins, including IRS-2, mSos-1, Shc, p85 subunit of phosphatidylinositol 3-kinase, SHP-2, Raf-1, and JAK2, were not associated with the INR beta-Myc.IRS-1 complex. Overexpression of INR beta-Myc and betaDelta kinases conferred an equivalent increase in cell proliferation in both 293 and Madin-Darby canine kidney cells, indicating that this growth response is independent of the carboxyl-terminal extension. However, INR beta-Myc-expressing cells exhibited enhanced survival relative to parental and betaDelta cells, suggesting that the carboxyl-terminal extension, through its interaction with IRS 1, plays a role in the regulation of cell death. PMID- 10455178 TI - Cys(577) is a conformationally mobile residue in the ATP-binding domain of the Na,K-ATPase alpha-subunit. AB - 2-[4'-Maleimidylanilino]naphthalene 6-sulfonic acid (MIANS) irreversibly inactivates Na,K-ATPase in a time- and concentration-dependent manner. Inactivation is prevented by 3 mM ATP or low K(+) (<1 mM); the protective effect K(+) is reversed at higher concentrations. This biphasic effect was also observed with K(+) congeners. In contrast, Na(+) ions did not protect. MIANS inactivation disrupted high affinity ATP binding. Tryptic fragments of MIANS-labeled protein were analyzed by reversed phase high performance liquid chromatography. ATP clearly protected one major labeled peptide peak. This observation was confirmed by separation of tryptic peptides in SDS-polyacrylamide gel electrophoresis revealing a single fluorescently-labeled peptide of approximately 5 kDa. N terminal amino acid sequencing identified the peptide (V(545)LGFCH...). This hydrophobic peptide contains only two Cys residues in all sodium pump alpha subunit sequences and is found in the major cytoplasmic loop between M4 and M5, a region previously associated with ATP binding. Subsequent digestion of the tryptic peptide with V8 protease and N-terminal amino acid sequencing identified the modified residue as Cys(577). The cation-dependent change in reactivity of Cys(577) implies structural alterations in the ATP-binding domain following cation binding and occlusion in the intramembrane domain of Na,K-ATPase and expands our knowledge of the extent to which cation binding and occlusion are sensed in the ATP hydrolysis domain. PMID- 10455179 TI - Molecular characterization of a novel basement membrane-associated proteoglycan, leprecan. AB - A monoclonal antibody was used in early studies to identify a novel chondroitin sulfate proteoglycan, secreted by L-2 cells, the core protein of which was approximately 100 kDa. To characterize this proteoglycan core protein at the molecular level, an L-2 cell cDNA library was probed by expression screening and solution hybridization. Northern blot analysis assigned transcript size to approximately 3.1 kilobases and, after contig assembly, the coding region of the mRNA corresponded to 2.18 kilobases. Immunoassays were performed to confirm the identity of this sequence, using a polyclonal antibody raised against an expressed fusion protein encoded by sequence representing the carboxyl half of the molecule. The antibody recognized the core protein in Western blots after prior digestion of the intact proteoglycan with chondroitinase ABC. Immunostaining tissue sections with the same antibody localized the proteoglycan to basement membranes, and expression of the entire sequence in Chinese hamster ovary K-1 cells showed that the protein encoded by the sequence secreted as a chondroitin sulfate proteoglycan. The core protein not only has motifs permitting glycosylation as a proteoglycan, but also possesses the endoplasmic reticulum retrieval signal, KDEL, which suggests that, in addition to its role as a basement membrane component, it may also participate in the secretory pathway of cells. PMID- 10455180 TI - New ether-a-go-go K(+) channel family members localized in human telencephalon. AB - A cDNA encoding a novel voltage-gated K(+) channel protein was isolated from human brain. This protein, termed BEC1, is 46% identical to rat elk in the ether a-go-go K(+) channel family. The BEC1 gene maps to the 12q13 region of the human genome. Northern blot analysis indicates that BEC1 is exclusively expressed in human brain, where the expression is concentrated in the telencephalic areas such as the cerebral cortex, amygdala, hippocampus, and striatum. By in situ hybridization, BEC1 is detected in the CA1-CA3 pyramidal cell layers and the dentate gyrus granule cell layers of the hippocampus. Specific signals are also found in neocortical neurons. Transfection of mammalian L929 and Chinese hamster ovary cells with BEC1 cDNA induces a voltage-gated outward current with a fast inactivation component. This current is insensitive to tetraethylammonium and quinidine. Additionally, a second related gene BEC2 was isolated from human brain. BEC2 is also brain-specific, located in the neocortex and the striatum, and functional as a channel gene. Phylogenetic analysis indicates that BEC1 and BEC2 constitute a subfamily, together with elk, in the ether-a-go-go family. The two genes may be involved in cellular excitability of restricted neurons in the human central nervous system. PMID- 10455181 TI - Two modes of PriA binding to DNA. AB - The role of PriA, required for the assembly of the phiX174-type primosome on DNA, in cellular DNA replication has been unclear since its discovery. Recent evidence, based on the phenotypes of strains carrying priA null mutations, has led to proposals that the primosome assembly activity of PriA was required to load replication forks at intermediates such as D loops during homologous recombination. McGlynn et al. (McGlynn, P., Al-Deib, A. A., Liu, J., Marians, K. J., and Lloyd, R. G. (1997) J. Mol. Biol. 270, 212-221) demonstrated that PriA could, in fact, bind D loops. We show here that there are two modes of stable binding of PriA to DNA. One mode, in which the enzyme binds 3'-single-stranded extensions from duplex DNAs, presumably reflects the 3' --> 5' DNA helicase activity of PriA. The D loop DNA binding activity of PriA can be accounted for by the second mode, where the enzyme binds bent DNA at three strand junctions. PMID- 10455182 TI - PriA-directed assembly of a primosome on D loop DNA. AB - Escherichia coli strains carrying null mutations in priA are chronically induced for the SOS response and are defective in homologous recombination, repair of UV damaged DNA, double-strand break repair, and both induced and constitutive stable DNA replication. This led to the proposal that PriA directed replication fork assembly at D loops formed by the homologous recombination machinery. The demonstration that PriA specifically recognized and bound D loop DNA supported this hypothesis. Using DNA footprinting as an assay, we show here that PriA also directs the assembly of a varphiX174-type primosome on D loop DNA. The ability to load a complete primosome on D loop DNA is a step necessary for replication fork assembly. PMID- 10455183 TI - Maximization of selenocysteine tRNA and U6 small nuclear RNA transcriptional activation achieved by flexible utilization of a Staf zinc finger. AB - Transcriptional activators Staf and Oct-1 play critical roles in the activation of small nuclear RNA (snRNA) and snRNA-type gene transcription. Recently, we established that Staf binding to the human U6 snRNA (hU6) and Xenopus selenocysteine tRNA (xtRNA(Sec)) genes requires different sets of the seven C2-H2 zinc fingers. In this work, using a combination of oocyte microinjection, electrophoretic mobility shift assays, and missing nucleoside experiments with wild-type and mutant promoters, we demonstrate that the hU6 gene requires zinc fingers 2-7 for Staf binding and Oct-1 for maximal transcriptional activity. In contrast, the xtRNA(Sec) gene needs the binding of the seven Staf zinc fingers, but not Oct-1, for optimal transcriptional capacity. Mutation in the binding site for Staf zinc finger 1 in the tRNA(Sec) promoter reduced both Staf binding and transcriptional activity. Conversely, introduction of a zinc finger 1 binding site in the hU6 promoter increased Staf binding but interfered with the simultaneous Staf and Oct-1 binding, thus reducing transcriptional activity. Collectively, these results show that the differential utilization of Staf zinc finger 1 represents a new, critical determinant of the transcriptional activation mechanism for the Xenopus tRNA(Sec) and human U6 snRNA genes. PMID- 10455184 TI - Tetratricopeptide repeat (TPR) motifs of p67(phox) participate in interaction with the small GTPase Rac and activation of the phagocyte NADPH oxidase. AB - The small GTPase Rac functions as a molecular switch in several important cellular events including cytoskeletal reorganization and activation of the phagocyte NADPH oxidase, the latter of which leads to production of superoxide, a precursor of microbicidal oxidants. During formation of the active oxidase complex at the membrane, the GTP-bound Rac appears to interact with the N terminal region of p67(phox), another indispensable activator that translocates from the cytosol upon phagocyte stimulation. Here we show that the p67(phox) N terminus lacks the CRIB motif, a well known Rac target, but contains four tetratricopeptide repeat (TPR) motifs with highly alpha-helical structure. Disruption of any of the N-terminal three TPRs, but the last one, results in defective interaction with Rac, while all the four are required for the NADPH oxidase activation. We also find that Arg-102 in the third repeat is likely involved in binding to Rac via an ionic interaction, and that replacement of this residue with Glu completely abrogates the capability of activating the oxidase both in vivo and in vitro. Thus the TPR motifs of p67(phox) are packed to function as a Rac target, thereby playing a crucial role in the active oxidase complex formation. PMID- 10455185 TI - Precursor processing of pro-ISG15/UCRP, an interferon-beta-induced ubiquitin-like protein. AB - Induction of the 17-kDa ubiquitin-like protein ISG15/UCRP and its subsequent conjugation to cellular targets is the earliest response to type I interferons. The polypeptide is synthesized as a precursor containing a carboxyl-terminal extension whose correct processing is required for subsequent ligation of the exposed mature carboxyl terminus. Recombinant pro-ISG15 is processed in extracts of human lung fibroblasts by a constitutive 100-kDa enzyme whose activity is unaffected by type I interferon stimulation. The processing enzyme has been purified to apparent homogeneity by a combination of ion exchange and hydrophobic chromatography and found to be stimulated 12-fold by micromolar concentrations of ubiquitin. Analysis of the products of pro-ISG15 processing enzyme demonstrates specific cleavage exclusively at the Gly(157)-Gly(158) peptide bond to generate a mature ISG15 carboxyl terminus. Irreversible inhibition of pro-ISG15 processing activity by thiol-specific alkylating agents and a pH rate dependence conforming to titration of a single group of pK(a) 8.1 indicate the 100-kDa enzyme is a thiol protease. Partial sequencing of a trypsin-derived peptide indicates the enzyme is either the human ortholog of yeast Ubp1 or a Ubp1-related protein. As yeast do not contain ISG15, these results suggest that a ubiquitin-specific enzyme was recruited for pro-ISG15/UCRP processing by adaptive divergence. PMID- 10455186 TI - Characterization of dTDP-4-dehydrorhamnose 3,5-epimerase and dTDP-4 dehydrorhamnose reductase, required for dTDP-L-rhamnose biosynthesis in Salmonella enterica serovar Typhimurium LT2. AB - The thymidine diphosphate-L-rhamnose biosynthesis pathway is required for assembly of surface glycoconjugates in a growing list of bacterial pathogens, making this pathway a potential therapeutic target. However, the terminal reactions have not been characterized. To complete assignment of the reactions, the four enzymes (RmlABCD) that constitute the pathway in Salmonella enterica serovar Typhimurium LT2 were overexpressed. The purified RmlC and D enzymes together catalyze the terminal two steps involving NAD(P)H-dependent formation of dTDP-L-rhamnose from dTDP-6-deoxy-D-xylo-4-hexulose. RmlC was assigned as the thymidine diphosphate-4-dehydrorhamnose 3,5-epimerase by showing its activity to be NAD(P)H-independent. Spectrofluorometric and radiolabeling experiments were used to demonstrate the ability of RmlC to catalyze the formation of dTDP-6-deoxy L-lyxo-4-hexulose from dTDP-6-deoxy-D-xylo-4-hexulose. Under reaction conditions, RmlC converted approximately 3% of its substrate to product. RmlD was unequivocally identified as the thymidine diphosphate-4-dehydrorhamnose reductase. The reductase property of RmlD was shown by equilibrium analysis and its ability to enable efficient biosynthesis of dTDP-L-rhamnose, even in the presence of low amounts of dTDP-6-deoxy-L-lyxo-4-hexulose. Comparison of 23 known and predicted RmlD sequences identified several conserved amino acid residues, especially the serine-tyrosine-lysine catalytic triad, characteristic for members of the reductase/epimerase/dehydrogenase protein superfamily. In conclusion, RmlD is a novel member of this protein superfamily. PMID- 10455187 TI - Insulin signaling is inhibited by micromolar concentrations of H(2)O(2). Evidence for a role of H(2)O(2) in tumor necrosis factor alpha-mediated insulin resistance. AB - Both hyperglycemia and tumor necrosis factor alpha (TNFalpha) were found to induce insulin resistance at the level of the insulin receptor (IR). How this effect is mediated is, however, not understood. We investigated whether oxidative stress and production of hydrogen peroxide could be a common mediator of the inhibitory effect. We report here that micromolar concentrations of H(2)O(2) dramatically inhibit insulin-induced IR tyrosine phosphorylation (pretreatment with 500 microM H(2)O(2) for 5 min inhibits insulin-induced IR tyrosine phosphorylation to 8%), insulin receptor substrate 1 phosphorylation, as well as insulin downstream signaling such as activation of phosphatidylinositol 3-kinase (inhibited to 57%), glucose transport (inhibited to 36%), and mitogen-activated protein kinase activation (inhibited to 7.2%). Both sodium orthovanadate, a selective inhibitor of tyrosine-specific phosphatases, as well as the protein kinase C inhibitor Go6976 reduced the inhibitory effect of hydrogen peroxide on IR tyrosine phosphorylation. To investigate whether H(2)O(2) is involved in hyperglycemia- and/or TNFalpha-induced insulin resistance, we preincubated the cells with the H(2)O(2) scavenger catalase prior to incubation with 25 mM glucose, 25 mM 2-deoxyglucose, 5.7 nM TNFalpha, or 500 microM H(2)O(2), respectively, and subsequent insulin stimulation. Whereas catalase treatment completely abolished the inhibitory effect of H(2)O(2) and TNFalpha on insulin receptor autophosphorylation, it did not reverse the inhibitory effect of hyperglycemia. In conclusion, these results demonstrate that hydrogen peroxide at low concentrations is a potent inhibitor of insulin signaling and may be involved in the development of insulin resistance in response to TNFalpha. PMID- 10455188 TI - Three isoforms of mammalian hyaluronan synthases have distinct enzymatic properties. AB - Three mammalian hyaluronan synthase genes, HAS1, HAS2, and HAS3, have recently been cloned. In this study, we characterized and compared the enzymatic properties of these three HAS proteins. Expression of any of these genes in COS-1 cells or rat 3Y1 fibroblasts yielded de novo formation of a hyaluronan coat. The pericellular coats formed by HAS1 transfectants were significantly smaller than those formed by HAS2 or HAS3 transfectants. Kinetic studies of these enzymes in the membrane fractions isolated from HAS transfectants demonstrated that HAS proteins are distinct from each other in enzyme stability, elongation rate of HA, and apparent K(m) values for the two substrates UDP-GlcNAc and UDP-GlcUA. Analysis of the size distributions of hyaluronan generated in vitro by the recombinant proteins demonstrated that HAS3 synthesized hyaluronan with a molecular mass of 1 x 10(5) to 1 x 10(6) Da, shorter than those synthesized by HAS1 and HAS2 which have molecular masses of 2 x 10(5) to approximately 2 x 10(6) Da. Furthermore, comparisons of hyaluronan secreted into the culture media by stable HAS transfectants showed that HAS1 and HAS3 generated hyaluronan with broad size distributions (molecular masses of 2 x 10(5) to approximately 2 x 10(6) Da), whereas HAS2 generated hyaluronan with a broad but extremely large size (average molecular mass of >2 x 10(6) Da). The occurrence of three HAS isoforms with such distinct enzymatic characteristics may provide the cells with flexibility in the control of hyaluronan biosynthesis and functions. PMID- 10455189 TI - Cytoskeleton-dependent tyrosine phosphorylation of the p130(Cas) family member HEF1 downstream of the G protein-coupled calcitonin receptor. Calcitonin induces the association of HEF1, paxillin, and focal adhesion kinase. AB - HEF1 is a recently described p130(Cas)-like docking protein that contains one SH3 domain and multiple SH2 binding motifs. In B cells, HEF1 is phosphorylated by a cytoskeleton-dependent mechanism that is triggered by integrin ligation. However, the induction of HEF1 phosphorylation by G protein-coupled receptors has not been reported. We found that HEF1, but not p130(Cas), is tyrosine-phosphorylated following stimulation of the rabbit C1a calcitonin receptor stably expressed in HEK-293 cells. The calcitonin-induced tyrosine phosphorylation of HEF1 increased in a time- and dose-dependent manner. Dibutyryl cAMP and forskolin had little or no effect on HEF1 phosphorylation, and the protein kinase A inhibitor H89 failed to detectably inhibit the response to calcitonin, indicating that the G(s)/cAMP/protein kinase A pathway does not mediate the calcitonin effect. Pertussis toxin, which selectively blocks G(i/o) signaling, also had no effect. Increasing cytosolic Ca(2+) with ionomycin stimulated HEF1 phosphorylation and preventing any calcitonin-induced change in cytosolic calcium by a combination of BAPTA and extracellular EGTA completely blocked the calcitonin-induced tyrosine phosphorylation of HEF1. Phorbol 12-myristate 13-acetate also induced HEF1 tyrosine phosphorylation, and the protein kinase C inhibitor calphostin C completely inhibited both calcitonin- and phorbol 12-myristate 13-acetate stimulated HEF1 phosphorylation. Calcitonin also induced the tyrosine phosphorylation of paxillin and focal adhesion kinase, and the association of these two proteins with HEF1. Pretreatment with cytochalasin D, which disrupts actin microfilaments, prevented the calcitonin-induced HEF1 and paxillin phosphorylation. In conclusion, the calcitonin-stimulated tyrosine phosphorylation of HEF1 is mediated by calcium- and protein kinase C-dependent mechanisms and requires the integrity of the actin cytoskeleton. PMID- 10455190 TI - Follicular thyroglobulin (TG) suppression of thyroid-restricted genes involves the apical membrane asialoglycoprotein receptor and TG phosphorylation. AB - Follicular thyroglobulin (TG) decreases expression of the thyroid-restricted transcription factors, thyroid transcription factor (TTF)-1, TTF-2, and Pax-8, thereby suppressing expression of the sodium iodide symporter, thyroid peroxidase, TG, and thyrotropin receptor genes (Suzuki, K., Lavaroni, S., Mori, A., Ohta, M., Saito, J., Pietrarelli, M., Singer, D. S., Kimura, S., Katoh, R., Kawaoi, A. , and Kohn, L. D. (1997) Proc. Natl. Acad. Sci. U. S. A. 95, 8251 8256). The ability of highly purified 27, 19, or 12 S follicular TG to suppress thyroid-restricted gene expression correlates with their ability to bind to FRTL 5 thyrocytes and is inhibited by a specific antibody to the thyroid apical membrane asialoglycoprotein receptor (ASGPR), which is related to the ASGPR of liver cells. Phosphorylating serine/threonine residues of TG, by autophosphorylation or protein kinase A, eliminates TG suppression and enhances transcript levels of the thyroid-restricted genes 2-fold in the absence of a change in TG binding to the ASGPR. Follicular TG suppression of thyroid restricted genes is thus mediated by the ASPGR on the thyrocyte apical membrane and regulated by a signal system wherein phosphorylation of serine/threonine residues on the bound ligand is an important component. These data provide a hitherto unsuspected role for the ASGPR in transcriptional signaling, aside from its role in endocytosis. They establish a functional role for phosphorylated serine/threonine residues on the TG molecule. PMID- 10455191 TI - Kinetic analysis of the actinorhodin aromatic polyketide synthase. AB - Type II polyketide synthases (PKSs) are bacterial multienzyme systems that catalyze the biosynthesis of a broad range of natural products. A core set of subunits, consisting of a ketosynthase, a chain length factor, an acyl carrier protein (ACP) and possibly a malonyl CoA:ACP transacylase (MAT) forms a "minimal" PKS. They generate a poly-beta-ketone backbone of a specified length from malonyl CoA derived building blocks. Here we (a) report on the kinetic properties of the actinorhodin minimal PKS, and (b) present further data in support of the requirement of the MAT. Kinetic analysis showed that the apoACP is a competitive inhibitor of minimal PKS activity, demonstrating the importance of protein protein interactions between the polypeptide moiety of the ACP and the remainder of the minimal PKS. In further support of the requirement of MAT for PKS activity, two new findings are presented. First, we observe hyperbolic dependence of PKS activity on MAT concentration, saturating at very low amounts (half maximal rate at 19.7 +/- 5.1 nM). Since MAT can support PKS activity at less than 1/100 the typical concentration of the ACP and ketosynthase/chain length factor components, it is difficult to rule out the presence of trace quantities of MAT in a PKS reaction mixture. Second, an S97A mutant was constructed at the nucleophilic active site of the MAT. Not only can this mutant protein support PKS activity, it is also covalently labeled by [(14)C]malonyl-CoA, demonstrating that the serine nucleophile (which has been the target of PMSF inhibition in earlier studies) is dispensible for MAT activity in a Type II PKS system. PMID- 10455192 TI - Hepatocyte nuclear factor 3 relieves chromatin-mediated repression of the alpha fetoprotein gene. AB - The alpha-fetoprotein gene (AFP) is tightly regulated at the tissue-specific level, with expression confined to endoderm-derived cells. We have reconstituted AFP transcription on chromatin-assembled DNA templates in vitro. Our studies show that chromatin assembly is essential for hepatic-specific expression of the AFP gene. While nucleosome-free AFP DNA is robustly transcribed in vitro by both cervical (HeLa) and hepatocellular (HepG2) carcinoma extracts, the general transcription factors and transactivators present in HeLa extract cannot relieve chromatin-mediated repression of AFP. In contrast, preincubation with either HepG2 extract or HeLa extract supplemented with recombinant hepatocyte nuclear factor 3 alpha (HNF3alpha), a hepatic-enriched factor expressed very early during liver development, is sufficient to confer transcriptional activation on a chromatin-repressed AFP template. Transient transfection studies illustrate that HNF3alpha can activate AFP expression in a non-liver cellular environment, confirming a pivotal role for HNF3alpha in establishing hepatic-specific gene expression. Restriction enzyme accessibility assays reveal that HNF3alpha promotes the assembly of an open chromatin structure at the AFP promoter. Combined, these functional and structural data suggest that chromatin assembly establishes a barrier to block inappropriate expression of AFP in non-hepatic tissues and that tissue-specific factors, such as HNF3alpha, are required to alleviate the chromatin-mediated repression. PMID- 10455193 TI - ATP-dependent activation of K(Ca) and ROMK-type K(ATP) channels in human submandibular gland ductal cells. AB - [Ca(2+)](i) and membrane current were measured in human submandibular gland ductal (HSG) cells to determine the regulation of salivary cell function by ATP. 1-10 microM ATP activated internal Ca(2+) release, outward Ca(2+)-dependent K(+) channel (K(Ca)), and inward store-operated Ca(2+) current (I(SOC)). The subsequent addition of 100 microM ATP activated an inwardly rectifying K(+) current, without increasing [Ca(2+)](i). The K(+) current was also stimulated by ATP in cells treated with thapsigargin in a Ca(2+)-free medium and was blocked by glibenclamide and tolbutamide, but not by charybdotoxin. This suggests the involvement of a Ca(2+)-independent, sulfonylurea-sensitive K(+) channel (K(ATP)). UTP mimicked the low [ATP] effects, while benzoyl-ATP activated internal Ca(2+) release, a Ca(2+) influx pathway, and K(Ca). Thus, ATP acts via P(2U) (P2Y(2)) and P(2Z) (P2X(7)) receptors to increase [Ca(2+)](i) and activate K(Ca), but not K(ATP). Importantly, (i) ROMK1 and the cystic fibrosis transmembrane regulator protein (but not SUR1, SUR2A, or SUR2B) and (ii) cAMP stimulated Cl(-) and K(+) currents were detected in HSG cells. These data demonstrate for the first time that a ROMK-type K(ATP) channel is present in salivary gland duct cells that is regulated by extracellular ATP and possibly by the cystic fibrosis transmembrane regulator. This reveals a potentially novel mechanism for K(+) secretion in these cells. PMID- 10455194 TI - Vascular endothelial growth factor signals endothelial cell production of nitric oxide and prostacyclin through flk-1/KDR activation of c-Src. AB - Vascular endothelial growth factor (VEGF) is a potent endothelial cell-specific mitogen that promotes angiogenesis, vascular hyperpermeability, and vasodilation by autocrine mechanisms involving nitric oxide (NO) and prostacyclin (PGI(2)) production. These experiments used immunoprecipitation and immunoassay procedures to characterize the signaling pathways by which VEGF induces NO and PGI(2) formation in cultured endothelial cells. The data showed that VEGF stimulates complex formation of the flk-1/kinase-insert domain-containing receptor (KDR) VEGF receptor with c-Src and that Src activation is required for VEGF induction of phospholipase C gamma1 activation and inositol 1,4,5-trisphosphate formation. Reporter cell assays showed that VEGF promotes a approximately 50-fold increase in NO formation, which peaks at 5-20 min. This effect is mediated by a signaling cascade initiated by flk-1/KDR activation of c-Src, leading to phospholipase C gamma1 activation, inositol 1,4,5-trisphosphate formation, release of [Ca(2+)](i) and nitric oxide synthase activation. Immunoassays of VEGF-induced 6-keto prostaglandin F(1alpha) formation as an indicator of PGI(2) production revealed a 3-4-fold increase that peaked at 45-60 min. The PGI(2) signaling pathway follows the NO pathway through release of [Ca(2+)](i), but diverges prior to NOS activation and also requires activation of mitogen-activated protein kinase. These results suggest that NO and PGI(2) function in parallel in mediating the effects of VEGF. PMID- 10455195 TI - Enzymatic repair of 5-formyluracil. I. Excision of 5-formyluracil site specifically incorporated into oligonucleotide substrates by alka protein (Escherichia coli 3-methyladenine DNA glycosylase II). AB - 5-Formyluracil (fU) is a major thymine lesion produced by reactive oxygen radicals and photosensitized oxidation. We have previously shown that fU is a potentially mutagenic lesion due to its elevated frequency to mispair with guanine. Therefore, fU can exist in DNA as a correctly paired fU:A form or an incorrectly paired fU:G form. In this work, fU was site-specifically incorporated opposite A in oligonucleotide substrates to delineate the cellular repair mechanism of fU paired with A. The repair activity for fU was induced in Escherichia coli upon exposure to N-methyl-N'-nitro-N-nitrosoguanidine, and the induction was dependent on the alkA gene, suggesting that AlkA (3-methyladenine DNA glycosylase II) was responsible for the observed activity. Activity assay and determination of kinetic parameters using purified AlkA and defined oligonucleotide substrates containing fU, 5-hydroxymethyluracil (hU), or 7 methylguanine (7mG) revealed that fU was recognized by AlkA with an efficiency comparable to that of 7mG, a good substrate for AlkA, whereas hU, another major thymine methyl oxidation products, was not a substrate. (1)H and (13)C NMR chemical shifts of 5-formyl-2'-deoxyuridine indicated that the 5-formyl group caused base C-6 and sugar C-1' to be electron deficient, which was shown to result in destabilization of the N-glycosidic bond. These features are common in other good substrates for AlkA and are suggested to play key roles in the differential recognition of fU, hU, and intact thymine. Three mammalian repair enzymes for alkylated and oxidized bases cloned so far (MPG, Nth1, and OGG1) did not recognize fU, implying that the mammalian repair activity for fU resided on a yet unidentified protein. In the accompanying paper (Terato, H., Masaoka, A., Kobayashi, M., Fukushima, S., Ohyama, Y., Yoshida, M., and Ide, H., J. Biol. Chem. 274, 25144-25150), possible repair mechanisms for fU mispaired with G are reported. PMID- 10455196 TI - Enzymatic repair of 5-formyluracil. II. Mismatch formation between 5-formyluracil and guanine during dna replication and its recognition by two proteins involved in base excision repair (AlkA) and mismatch repair (MutS). AB - 5-Formyluracil (fU), a major methyl oxidation product of thymine, forms correct (fU:A) and incorrect (fU:G) base pairs during DNA replication. In the accompanying paper (Masaoka, A., Terato, H., Kobayashi, M., Honsho, A., Ohyama, Y., and Ide, H. (1999) J. Biol. Chem. 274, 25136-25143), it has been shown that fU correctly paired with A is recognized by AlkA protein (Escherichia coli 3 methyladenine DNA glycosylase II). In the present work, mispairing frequency of fU with G and cellular repair protein that specifically recognized fU:G mispairs were studied using defined oligonucleotide substrates. Mispairing frequency of fU was determined by incorporation of 2'-deoxyribonucleoside 5'-triphosphate of fU opposite template G using DNA polymerase I Klenow fragment deficient in 3'-5' exonuclease. Mispairing frequency of fU was dependent on the nearest neighbor base pair in the primer terminus and 2-12 times higher than that of thymine at pH 7.8 and 2.6-6.7 times higher at pH 9.0 with an exception of the nearest neighbor T(template):A(primer). AlkA catalyzed the excision of fU placed opposite G, as well as A, and the excision efficiencies of fU for fU:G and fU:A pairs were comparable. In addition, MutS protein involved in methyl-directed mismatch repair also recognized fU:G mispairs and bound them with an efficiency comparable to T:G mispairs, but it did not recognize fU:A pairs. Prior complex formation between MutS and a heteroduplex containing an fU:G mispair inhibited the activity of AlkA to fU. These results suggest that fU present in DNA can be restored by two independent repair pathways, i.e. the base excision repair pathway initiated by AlkA and the methyl-directed mismatch repair pathway initiated by MutS. Biological relevance of the present results is discussed in light of DNA replication and repair in cells. PMID- 10455197 TI - Mutational and pH studies of the 3' --> 5' exonuclease activity of bacteriophage T4 DNA polymerase. AB - The 3' --> 5' exonuclease activity of proofreading DNA polymerases requires two divalent metal ions, metal ions A and B. Mutational studies of the 3' --> 5' exonuclease active center of the bacteriophage T4 DNA polymerase indicate that residue Asp-324, which binds metal ion A, is the single most important residue for the hydrolysis reaction. In the absence of a nonenzymatic source of hydroxide ions, an alanine substitution for residue Asp-324 reduced exonuclease activity 10 100-fold more than alanine substitutions for the other metal-binding residues, Asp-112 and Asp-219. Thus, exonuclease activity is reduced 10(5)-fold for the D324A-DNA polymerase compared with the wild-type enzyme, while decreases of 10(3) to 10(4)-fold are detected for the D219A- and D112A/E114A-DNA polymerases, respectively. Our results are consistent with the proposal that a water molecule, coordinated by metal ion A, forms a metal-hydroxide ion that is oriented to attack the phosphodiester bond at the site of cleavage. Residues Glu-114 and Lys 299 may assist the reaction by lowering the pK(a) of the metal ion-A coordinated water molecule, whereas residue Tyr-320 may help to reorient the DNA from the binding conformation to the catalytically active conformation. PMID- 10455198 TI - Properties of a cyclodextrin-specific, unusual porin from Klebsiella oxytoca. AB - The function of CymA, 1 of the 10 gene products involved in cyclodextrin uptake and metabolism by Klebsiella oxytoca, was characterized. CymA is essential for growth on cyclodextrins, but it can also complement the deficiency of a lamB (maltoporin) mutant of Escherichia coli for growth on linear maltodextrins, indicating that both cyclic and linear oligosaccharides are accepted as substrates. CymA was overproduced in E. coli and purified to apparent homogeneity. CymA is a component of the outer membrane, is processed from a signal peptide-containing precursor, and possesses a high content of antiparallel beta-sheet. Incorporation of CymA into lipid bilayers and conductance measurements revealed that it forms ion-permeable channels, which exhibit a substantial current noise. CymA-induced membrane conductance decreased considerably upon addition of alpha-cyclodextrin. Titration experiments allowed the calculation of a half-saturation constant, K(S), of 28 microM for its binding to CymA. CymA assembled in vitro to two-dimensionally crystalline tubular membranes, which, on electron microscopy, are characterized by a p1-related two sided plane group. The crystallographic unit cell contains four monomeric CymA molecules showing a central pore. The lattice parameters are a = 16.1 nm, b = 3.8 nm, gamma = 93 degrees. CymA does not form trimeric complexes in lipid membranes and shows no tendency to trimerize in solution. CymA thus is an atypical porin with novel properties specialized to transfer cyclodextrins across the outer membrane. PMID- 10455199 TI - Elastase-mediated release of heparan sulfate proteoglycans from pulmonary fibroblast cultures. A mechanism for basic fibroblast growth factor (bFGF) release and attenuation of bfgf binding following elastase-induced injury. AB - We have investigated elastase-mediated alterations in the expression of basic fibroblast growth factor (bFGF) receptors and proteoglycan co-receptors and characterized the subsequent effects on bFGF receptor binding profiles. For these studies, pulmonary fibroblast cultures were treated with porcine pancreatic elastase, and elastase-mediated changes in bFGF receptor expression and binding profiles were assessed. Quantitation of [(35)S]sulfate-labeled proteoglycan and total glycosaminoglycan release from fibroblast matrices indicated that elastase treatment released sulfated proteoglycan from the cell surface in a time- and dose-dependent fashion that correlated strongly with elastase-mediated bFGF release. Ligand binding studies indicated that elastase treatment decreased total binding of (125)I-bFGF to the cell surface and affected both fibroblast growth factor receptor and heparan sulfate proteoglycan (HSPG) binding sites. Western blot analyses indicated that elastase treatment did not release significant amounts of fibroblast growth factor receptor protein. These findings indicate that elastase-mediated HSPG release from fibroblast matrices reduces the effective affinity of bFGF for its receptor. Collectively, these studies suggest that HSPG co-receptors are important mediators of the pulmonary fibroblast response to elastase treatment and that bFGF, HSPG, and other elastase-released entities play an important role in the response of the lung to chronic injury. PMID- 10455200 TI - Neuronal sensitivity to tetanus toxin requires gangliosides. AB - Tetanus toxin produces spastic paralysis in situ by blocking inhibitory neurotransmitter release in the spinal cord. Although di- and trisialogangliosides bind tetanus toxin, their role as productive toxin receptors remains unclear. We examined toxin binding and action in spinal cord cell cultures grown in the presence of fumonisin B(1), an inhibitor of ganglioside synthesis. Mouse spinal cord neurons grown for 3 weeks in culture in 20 microM fumonisin B(1) develop dendrites, axons, and synaptic terminals similar to untreated neurons, even though thin layer chromatography shows a greater than 90% inhibition of ganglioside synthesis. Absence of tetanus and cholera toxin binding by toxin-horseradish peroxidase conjugates or immunofluorescence further indicates loss of mono- and polysialogangliosides. In contrast to control cultures, tetanus toxin added to fumonisin B(1)-treated cultures does not block potassium-stimulated glycine release, inhibit activity-dependent uptake of FM1 43, or abolish immunoreactivity for vesicle-associated membrane protein, the toxin substrate. Supplementing fumonisin B(1)-treated cultures with mixed brain gangliosides completely restores the ability of tetanus toxin to bind to the neuronal surface and to block neurotransmitter release. These data demonstrate that fumonisin B(1) protects against toxin-induced synaptic blockade and that gangliosides are a necessary component of the receptor mechanism for tetanus toxin. PMID- 10455201 TI - Tic22 is targeted to the intermembrane space of chloroplasts by a novel pathway. AB - Tic22 previously was identified as a component of the general import machinery that functions in the import of nuclear-encoded proteins into the chloroplast. Tic22 is peripherally associated with the outer face of the inner chloroplast envelope membrane, making it the first known resident of the intermembrane space of the envelope. We have investigated the import of Tic22 into isolated chloroplasts to define the requirements for targeting of proteins to the intermembrane space. Tic22 is nuclear-endoded and synthesized as a preprotein with a 50-amino acid N-terminal presequence. The analysis of deletion mutants and chimerical proteins indicates that the precursor of Tic22 (preTic22) presequence is necessary and sufficient for targeting to the intermembrane space. Import of preTic22 was stimulated by ATP and required the presence of protease-sensitive components on the chloroplast surface. PreTic22 import was not competed by an excess of an authentic stromal preprotein, indicating that targeting to the intermembrane space does not involve the general import pathway utilized by stromal preproteins. On the basis of these observations, we conclude that preTic22 is targeted to the intermembrane space of chloroplasts by a novel import pathway that is distinct from known pathways that target proteins to other chloroplast subcompartments. PMID- 10455203 TI - Loss of heterozygosity analysis to diagnose adrenal cortical carcinoma: are we there yet?-reply PMID- 10455202 TI - Loss of heterozygosity analysis to diagnose adrenal cortical carcinoma: are we there yet? PMID- 10455204 TI - Observer variation in cytologic grading for cervical dysplasia of Papanicolaou smears with the PAPNET testing system. AB - BACKGROUND: To assess the interobserver and intraobserver variation of Papanicolaou (Pap) smear screening with the computer-assisted (neural network based) PAPNET Testing System in diagnosing cervical smear abnormalities, results of agreement were compared with the interobserver and intraobserver variation of conventional smear analysis. METHODS: Cervical smears obtained from women in 1996 were reevaluated both by conventional light microscopy and with use of the PAPNET Testing System by the same four investigators, and results were compared with the original screening diagnoses obtained by both methods. RESULTS: The interobserver results for epithelial abnormalities (the degree of agreement between the cytologists), characterized by weighted kappa statistics, were 0.71 (95% CI: 0. 68-0.73) for PAPNET screening and 0.69 (95% CI: 0.66-0.72) for conventional screening. No significant differences were found among the individual results obtained by the four cytotechnologists (intraobserver variation) with conventional screening versus PAPNET reviewing. CONCLUSIONS: Pap smear grading with the PAPNET Testing System has interobserver and intraobserver variation similar to that of conventional screening of Pap smears in routine use. Cancer (Cancer Cytopathol) PMID- 10455205 TI - Cervical Papanicolaou smear abnormalities in inner city Bronx adolescents: prevalence, progression, and immune modifiers. AB - BACKGROUND: The aim of this study was to quantify the prevalence of cervical smear abnormalities in sexually active adolescents and identify the effect of immune-modifying conditions. METHODS: Two hundred seventy-one females ages 13-22 years attending a clinic for sexually transmitted disease (STD) evaluation had cervical Papanicoloau (Pap) smears and completed sexual history questionnaires. Results of all follow-up Pap smears were obtained. Medical charts were available for 54 patients with cytologic follow-up and were reviewed for the presence of immune-modifying conditions. Follow-up smear results for patients with and without immune-modifying conditions were compared. Abnormality rates for all cervical smears seen in 1995 at Montefiore Medical Center were also obtained. RESULTS: The smear abnormality rate for adolescents was 20. 7% (abnormal squamous cells of undetermined significance [ASCUS], 12. 2%; low grade squamous intraepithelial lesion [LGSIL], 7.7%; high grade squamous intraepithelial lesion [HGSIL], 0.7%) compared with all adult females, for whom the rate was 13.2% (ASCUS, 9.9%; LGSIL, 2.5%; HGSIL, 0.6%; carcinoma 0.2%) (P < 0.0002). Of 20 initial ASCUS patients, 6 (30%) showed LGSIL or HGSIL on follow-up. Chart review allowed the clinical immune status of 54 patients to be determined. Of 14 patients with an immune-modifying condition (9 HIV positive patients, 3 receiving oral steroids, 1 liver transplant patient receiving steroids, and 1 with intestinal lymphangiectasia), 11 (78. 6%) developed or maintained an abnormality on cytologic follow-up. Of 40 patients with no identifiable immune-modifying condition, 11 (27.5%) developed or maintained an abnormality on cytologic follow up (P < 0.00082). CONCLUSIONS: Sexually active adolescents are at higher risk of developing a significant cervical smear abnormality, especially LGSIL. Patients with an atypical Pap smear or immune-modifying condition require more attentive gynecologic monitoring. Cancer (Cancer Cytopathol) PMID- 10455206 TI - Fine-needle aspiration cytology of hemangiopericytoma: A report of five cases. AB - BACKGROUND: Hemangiopericytoma (HPC) is a relatively rare neoplasm, accounting for approximately 2.5% of all soft tissue tumors. Its histopathology has been well documented but to the authors' knowledge reports regarding its fine-needle aspiration (FNA) cytology rarely are encountered. In the current study the authors report the cytologic findings in FNA specimens from nine confirmed cases of HPC and attempt to correlate the cytologic features with the biologic outcomes. METHODS: FNA was performed with or without radiologic guidance. Corresponding sections of tissue were reviewed in conjunction with the cytologic preparations. RESULTS: Nine FNAs were performed in 5 patients (3 men and 2 women) with an age range of 38-77 years (mean, 56 years). Two lesions were primary soft tissue lesions arising in the lower extremities; seven were recurrent or metastatic lesions from bone (one lesion), kidney (one lesion), pelvic fossa (one lesion), lower extremities (two lesions), trunk (one lesion), and breast (one lesion). All aspirates were cellular and were comprised of single and tightly packed clusters of oval to spindle-shaped cells aggregated around branched capillaries. Basement membrane material was observed in 6 cases (67%). The nuclei were uniform and oval, with finely granular chromatin and inconspicuous nucleoli in all cases except one. No mitotic figures or areas of necrosis were identified. A correct diagnosis of HPC was made on one primary lesion and all recurrent or metastatic lesions. CONCLUSIONS: HPCs show a spindle cell pattern in cytologic preparations and must be distinguished from more common spindle cell lesions. The presence of branched capillaries and abundant basement membrane material supports a diagnosis of HPC. Immunohistochemistry and electron microscopy performed on FNA samples may be helpful in the differential diagnosis. FNA is a useful and accurate tool with which to confirm recurrent or metastatic HPC; however, prediction of the biologic behavior of HPC based on cytologic features is not feasible. Cancer (Cancer Cytopathol) PMID- 10455207 TI - Cytopathology of insular carcinoma of the thyroid. AB - BACKGROUND: Insular carcinoma of the thyroid (ICT) first was reported in 1984. To the authors' knowledge, few cytology reports have been published since that time. The authors describe the cytologic features of six tissue-proven ICTs and propose criteria that suggest its diagnosis. METHODS: Four cases were thyroid fine-needle aspiration (FNA) samples. Two cases were FNAs of metastases. All cases were found to be classic ICT on examination of primary or metastatic surgical specimens. RESULTS: Three cases originally were diagnosed as carcinoma, including two FNAs of metastatic sites and one thyroid FNA. Two additional thyroid FNAs were diagnosed as suspicious for malignancy, favor follicular neoplasm. One case was termed a neoplasm, favor follicular type. Smears showed high cellularity and scanty colloid. Three cases were found to contain some microfollicles. One case showed a few papillae. Necrosis and mitosis were rare. Cells were round with pale, poorly defined cytoplasm. Nuclei were round and monomorphic with finely granular chromatin, mild hyperchromasia, smooth nuclear membranes, and small nucleoli. Nuclear grooves and inclusions were rare. CONCLUSIONS: Three cases were diagnosed as suspicious for follicular neoplasm, the main differential diagnosis of ICT. Both tumors exhibited high cellularity and scanty colloid. However, ICT showed a predominance of single cells whereas follicular neoplasms reveal microfollicles with more nuclear atypia. There is cytologic overlap between these two neoplasms. Papillary thyroid carcinoma should be distinguished from ICT easily because the latter usually does not reveal the classic cytologic features associated with the former. ICT should be considered in the differential diagnosis of follicular neoplasms. Features favoring ICT are predominance of single cells, small loose nests of cells with few microfollicles, and little nuclear atypia. Cancer (Cancer Cytopathol) PMID- 10455208 TI - Cytologic diagnosis and grading of ductal carcinoma in situ. AB - BACKGROUND: Fine-needle aspiration cytology plays an important role in the preoperative diagnosis of palpable masses as well as impalpable lesions that can only be sampled by stereotactic or ultrasound techniques. A further refinement of cytologic diagnosis would be the ability to distinguish among the different types of ductal carcinoma in situ (DCIS) also between in situ and invasive malignant disease. METHODS: Sixty-six cases of histologically proven, pure DCIS (39 high grade, 12 cribriform, and 15 low/intermediate grade) with a preoperative cytology report of carcinoma were retrieved from our files. All the cytology (wet-fixed and air-dried smears) was reviewed by G.M., and the histology sections were reviewed by G.T. Seven cytologic features, including cellularity, cell dissociation, nuclear size, cell uniformity, nucleoli, nuclear margins, and chromatin pattern, were assigned scores from 1 to 3. The presence of calcium, necrosis, and foamy macrophages was recorded. Cell clusters were examined for evidence of a cribriform pattern. Fat and stromal fragments were closely checked for infiltration by tumor cells. RESULTS: The cell type was predominantly large and pleomorphic in high grade DCIS, whereas it was mainly small and well differentiated in the cribriform and low grade types. Calcium and necrosis were seen in most high grade lesions and less frequently in the cribriform and low grade ones. Macrophages were more common in high grade and cribriform DCIS than in low/intermediate grade DCIS. Cribriform spaces were noted only in cribriform DCIS. Stromal and fat infiltration by tumor cells was not present in any of the aspirates. Myoepithelial cells were rarely seen. CONCLUSIONS: The presence of pleomorphic carcinoma cells, calcium, necrosis, and macrophages in the aspirate accompanied by "casting" calcification on the mammogram is virtually diagnostic of high grade (comedo) DCIS. Cribriform DCIS shows features of low grade carcinoma with a typical cribriform pattern of punched-out holes in the cell clusters. Low/intermediate grade DCIS has no particular distinguishing features. Cancer (Cancer Cytopathol) PMID- 10455209 TI - Telomerase activity: comparison between fine-needle aspiration and biopsy specimens for the detection of tumor cells. AB - BACKGROUND: The purpose of the current study was to determine telomerase activity as a sensitive biomarker for the detection of malignant cells in fine-needle aspiration (FNA) specimens. METHODS: FNA specimens with parallel samples of fresh tumor tissue were obtained from surgical specimens after surgical excision. Using a polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) assay, telomerase activity was determined systematically in FNA specimens (n = 21) and corresponding available tissue biopsy specimens (n = 16) containing malignant cells. In addition to a case of myelolipoma, normal counterparts for 3 of 16 cancer cases, including both biopsy and FNA specimens, also were available for the determination of telomerase activity. RESULTS: Telomerase activity was observed in 14 of 16 of the FNA specimens (88%) and 15 of 16 of the corresponding biopsy specimens (94%). Telomerase activity was detected in both the biopsy specimen and the corresponding FNA specimen, with one exception (a case of mucinous adenocarcinoma of the cecum). In contrast, specimens from three normal tissue biopsies and FNA specimens of normal tissue adjacent to the malignant lesions, as well as the myelolipoma, exhibited no telomerase activity. It is interesting to note that both tissue biopsy specimens and FNA specimens from a patient with high grade sarcoma were negative for telomerase activity. The examination of hematoxylin and eosin-stained adjacent tissue biopsy sections or FNA smears revealed similar low populations of lymphocytes, including those cases that were negative for telomerase activity. There was agreement in the detection of telomerase activity between tissue biopsies and their corresponding FNA specimens in 15 of the 16 patients, indicating a 94% concordance rate (95% confidence interval, 70%, 98%). CONCLUSIONS: The results of the current study clearly suggest that the telomerase activity in FNA specimens was comparable to that of their corresponding biopsy specimens, and that this activity was associated with the presence of malignant cells. The TRAP assay has potential for use in the detection of malignant cells in FNA specimens, particularly cases in which the cytology is not characteristically malignant and/or is present in insufficient numbers. Cancer (Cancer Cytopathol) PMID- 10455210 TI - Diagnosis and subclassification of follicle center and mantle cell lymphomas on fine-needle aspirates: A cytologic and immunocytochemical approach based on the Revised European-American Lymphoma (REAL) classification. AB - BACKGROUND: Cytologic distinction between follicle center lymphoma (FCL) and mantle cell lymphoma (MCL) is difficult with cytomorphology alone and requires immunophenotyping. The current study describes the distinction between follicle center and mantle cell lymphoma made with fine-needle aspiration (FNA) material. METHODS: One hundred ten cases primarily diagnosed and classified on FNA material as centroblastic-centrocytic (CBCC) and centrocytic (CC) non-Hodgkin lymphomas (NHLs) (Kiel classification) were included in the study. An additional retrospective immunocytochemical analysis was performed on frozen cytospin preparations using the monoclonal antibodies Bcl-2, CD10, CD5, CD23, CD43, and immunoglobulin M. RESULTS: The initial diagnostic workup classified 106 cases as CBCC-NHL and 4 as CC-NHL. The immunophenotype Bcl-2(+), CD10(+/-), CD5(-), CD23( /+), CD43(-) was observed in 93 of 106 previously reported CBCC NHLs. In 11 of 106 cytospin preparations, neoplastic B cells expressed the CD5 pan T marker and, as a group, showed the pattern Bcl (+/-), CD10(-/+), CD5(+), CD23(-), CD43(+), which is considered typical of MCL. Based on the additional immunocytochemical data, all but 2 of the tumors were reclassified as FCL (n = 93) and MCL (n = 15). The mean proliferation fraction measured by MIB-1 (Ki-67) immunoreactivity was 16.3% and 17.5% in FCL and MCL, respectively. The revised cytopathologic diagnosis correlated significantly (P < 10(-9)) with the histology of 65 patients who underwent surgical excision biopsy. CONCLUSIONS: Subclassification of follicle-derived low grade NHL can be established with high accuracy on FNA material if cytomorphology is corroborated by a complete immunophenotypic analysis, which can be performed on both fresh and frozen stored cytospin material. The currently used criteria can be applied to aspirated cells for a conclusive cytopathologic diagnosis of MCL, which is of great clinical importance. Cancer (Cancer Cytopathol) PMID- 10455211 TI - Bcl-2 expression on fine-needle aspirates from primary breast carcinoma: correlation with other biologic factors. AB - BACKGROUND: The bcl-2 gene encodes for a protein that is involved in cell death regulation. It frequently is expressed in breast tumors, in which it is associated with favorable prognostic factors. It has been suggested that bcl-2 also may act as a modulator of response to chemotherapy and/or endocrine therapy. Because fine-needle aspiration (FNA) biopsy has been established as a reliable method for the diagnosis and biologic characterization of breast carcinoma, we assessed Bcl-2 expression on FNAs from primary breast carcinoma and evaluated its correlations with other prognostic variables. METHODS: Bcl-2, estrogen receptor (ER), progesterone receptor (PgR), p53 protein expression, and Ki-67 growth fraction were evaluated by immunocytochemistry on FNAs from 130 patients with primary breast carcinoma. Nuclear cytologic grade was assessed on FNA smears. RESULTS: Bcl-2 was expressed in 99 of 130 FNAs (76%). Bcl-2 expression was correlated with positive ER (P < 0.001) and PgR (P < 0.001) status and inversely correlated with p53 (P = 0.0036), Ki-67 (P = 0.0073), and nuclear cytologic grade (P < 0.001). CONCLUSIONS: Bcl-2 expression, evaluated by immunocytochemistry on FNAs from primary breast carcinoma, correlates with favorable prognostic features such as ER and PgR expression, p53 negativity, a low Ki-67 index, and high tumor differentiation. These results are in agreement with those found on histologic samples. As FNA biopsy is used increasingly as a primary tool in the diagnosis of breast carcinoma, Bcl-2 evaluation by immunocytochemistry on FNA may provide, in addition to other biologic variables, useful information for prognostic and predictive purposes, particularly in patients considered to be candidates for neoadjuvant treatments. Cancer (Cancer Cytopathol) PMID- 10455212 TI - Utilization of microdissection and the polymerase chain reaction for the diagnosis of adrenal cortical carcinoma in fine-needle aspiration cytology. AB - BACKGROUND: Loss of heterozygosity (LOH) for several tumor suppressor genes (including loci on 3p, 1p, and 17p,) has been documented in surgical specimens of adrenal cortical carcinomas (ACCA) without accompanying losses in benign hyperplastic and adenomatous adrenal cortical lesions (ACL). This disparate pattern of LOH raises the possibility of exploitation of these differences for diagnostic utilization. Cytologic differentiation of benign versus malignant ACL may be impossible based solely on fine-needle aspiration (FNA) material. The authors attempted to extrapolate the genetic findings on surgical specimens to FNA specimens of ACL to determine whether LOH studies could be utilized as a definitive diagnostic tool. METHODS: Microdissection of archival material was performed on FNAs of ten ACCAs (stained with the Papanicolaou and Diff-Quik stains) with corresponding histologic material (stained with hematoxylin and eosin), one FNA of a benign ACL, and three touch preparations of benign adrenal cortex. LOH analysis was performed by polymerase chain reaction (PCR) with flanking markers for the following putative tumor suppressor genes: p53 (17p13; TP53), 1p (1p36; D1S165), and the von Hippel-Lindau gene at 3p25 (D3S1038 and D3S1110). RESULTS: Similar results were obtained with cytologic and histologic material. As expected, benign ACL showed no LOH for the markers examined. Of the informative ACCA cases, 70% showed LOH for at least 1 of the 3 markers tested on both FNA and histologic samples. For all cases with amplifiable DNA, there was a 100% concordance rate for LOH between cytologic and histologic material, with at least 7 of the 10 cytologic samples originating from metastatic lesions and all of the surgical material originating from the primary adrenal neoplasm. CONCLUSIONS: The results of this study suggest that the combination of microdissection and PCR for LOH of p53, 1p, and 3p25 from FNA material has the potential to be utilized to distinguish ACCA from benign ACL in informative cases. It also shows a 100% concordance rate between metastatic and primary ACCAs for the losses observed, a finding that can be extremely useful for the definitive identification of metastatic lesions. Archival cytologic preparations of ACCA are a reliable source of DNA for LOH studies. [See editorial counterpoint on pages 173-5 and reply to counterpoint on pages 176-7, this issue.] Cancer (Cancer Cytopathol) PMID- 10455213 TI - The Tall cell variant of papillary carcinoma of the thyroid: cytologic features and loss of heterozygosity of metastatic and/or recurrent neoplasms and primary neoplasms. AB - BACKGROUND: The Tall cell variant of papillary carcinoma of the thyroid (TCV) is characterized by the proliferation of oxyphilic, tall, columnar cells with a height-to-width ratio of at least 2:1. TCV exhibits more aggressive clinical behavior than conventional thyroid papillary carcinoma (CPC). Cytologic features suggestive of TCV have been described in fine-needle aspiration material from primary tumors. Similarly, loss of heterozygosity (LOH) for chromosome 1 (D1S243) and the p53 gene (TP53) have been reported in TCV but not in CPC, thus making exploitation of this genetic feature a potential tool for molecular discrimination between these two neoplasms. METHODS: Cytology samples of metastatic and/or recurrent neoplasms (M/R) (12 cases) and 7 cases of primary TCV obtained from 12 patients were evaluated. The cytologic findings of these cases were compared with previously published findings. Microdissection and polymerase chain reaction for LOH for chromosome 1 and p53 (D1S243 and TP53 markers) were performed on cytologic smears from 6 cases of M/R tumors and 3 cases of primary tumors. RESULTS: More then 50% of M/R showed atypical follicular cells with enlarged nuclei, granular chromatin, nuclear grooves, pseudoinclusions, and abundant finely granular cytoplasm. Cells were disposed in monolayers (58%) and papillary clusters (50%). Similar findings were present in cases of primary TCV. LOH studies showed that 4 of 6 M/R were noninformative and 2 of 3 cases of primary TCV were informative for the D1S243 marker; however, in contrast with previously published reports, no LOH was detected for the markers evaluated. CONCLUSIONS: M/R and primary TCV have similar cytologic features. Additional studies of larger series of M/R and primary TCV should be performed to delineate further any potential application of LOH for chromosome 1 and the p53 gene as a tool for diagnosing TCV with cytologic preparations. Cancer (Cancer Cytopathol) PMID- 10455214 TI - The Lancer Ethnicity Scale. PMID- 10455215 TI - Reply. PMID- 10455216 TI - Eyelid resurfacing. AB - BACKGROUND AND OBJECTIVE: Laser resurfacing of eyelids was examined in a series of experiments designed to measure beam parameters, surface temperatures, ablation characteristics, thermal damage, tissue responses and clinical outcomes. These data were collected for the purpose of developing a logical basis for clinical dosimetry. STUDY DESIGN: All experiments were conducted with similar short-pulse CO(2) lasers (TruPulse, Albuquerque, NM) where the beam had been carefully characterized and calibrated. The chronological sequence examined begins with the photophysical laser/tissue interactions during the first few microsec of irradiation and ends with an evaluation of the efficacy of wrinkle reduction nine months after treatment. RESULTS: Eyelid tissue removed by the first and second passes consisted mostly of epidermis with about 38 microm of thermal damage into the papillary dermis. Erythema resolved within four weeks and most patients experienced 70-100% wrinkle reduction by nine months. CONCLUSION: A layer of contracted dermal scar tissue that replaced the thermally challenged zone in the dermis is identified as the substrate for wrinkle reduction. The data support the following dosimetry for periorbital wrinkle reduction: One pass 4-6 J/cm(2) (350-500 mJ into a 3 x 3 mm spot). A second treatment after 9-12 months may be more beneficial than a second pass. PMID- 10455217 TI - Use of the Q-switched alexandrite laser (755 nm, 100 nsec) for eyebrow tattoo removal. AB - BACKGROUND AND OBJECTIVE: Permanent tattooing for cosmetic reasons has increased in recent years; as a consequence, there has been an increase of requests for pigment removal due to complications or undesired results. The Q-switched alexandrite laser has been found useful in removing black exogenous pigment, which is the most popular color in eyebrow enhancement. We report the case of a patient with black-pigment eyebrow cosmetic tattoo after treatment with the Q switched alexandrite laser. STUDY DESIGN/MATERIALS AND METHODS: Treatment conditions included 755-nm wavelength, 100 +/- 10-nsec pulse width, and 3-mm spot size. Fluence threshold was determined, and a spot test was made at the first visit. Single impact technique with 10% overlapping was applied to the whole tattoo. Five treatments were performed with a mean fluence of 7 J/cm(2). RESULTS: Complete pigment removal was achieved after five sessions. Superficial bleeding and vesicle formation was observed. CONCLUSIONS: Eyebrow tattooing can be treated efficiently with the use of the Q-switched alexandrite laser when black pigment has been used for cosmetic reasons. PMID- 10455218 TI - Total intravenous anesthesia for office-based laser facial resurfacing. AB - BACKGROUND AND OBJECTIVE: Providing general anesthesia in an office-based setting can be time consuming, risky, and expensive. The purpose of this study was to describe a technique for total intravenous anesthesia (TIVA) technique that can be easily utilized by anesthesiologists in an office-based setting for laser facial resurfacing. STUDY DESIGN/MATERIALS AND METHODS: Twenty-five American Society of Anesthesiologists (ASA) status classification I-II patients (22 females and three males) elected general anesthesia for laser facial resurfacing. All patients were premedicated with glycopyrrolate (0.2 mg IV). All anesthetics were administered by board-certified anesthesiologists, and ASA Standards for Anesthesia Monitoring were strictly followed. An induction dose of propofol (2. 0 2.5 mg/kg IV) was followed by laryngeal mask airway insertion (size 3 or 4). TIVA was maintained with a propofol infusion (50-250 mcg/kg/minute IV). Supplemental midazolam (2-4 mg IV), fentanyl (0. 05-0.20 mg IV), and oxygen (2-4 l/minute) were administered as needed. After completion of the laser procedure, TIVA was discontinued and the patients were allowed to awaken. Patients were discharged after achieving a Modified Post-Anesthetic Discharge Score of >/= 9. RESULTS: Mean procedure duration was 48 +/- 21 minutes, and time to discharge after the procedure was 16 +/-6 minutes. All procedures and anesthetics were well tolerated and without complications. The only post-procedure complaint was an isolated, minor, and temporary sore throat. CONCLUSIONS: TIVA is an excellent method for providing anesthesia for laser facial resurfacing in an office-based setting. PMID- 10455219 TI - Simulation of color of port wine stain skin and its dependence on skin variables. AB - BACKGROUND AND OBJECTIVE: The understanding of why Port Wine Stain (PWS) skin is redder and darker as compared to normal skin has so far been based on qualitative analysis. This study aims at quantitatively analyzing the influence of skin anatomy variables on perceived skin color. STUDY DESIGN/MATERIALS AND METHODS: Reflectance spectra for visible light from normal and Port Wine Stain skin have been calculated using a Monte Carlo algorithm applied to a multi-layered skin model. Skin parameters that were varied are pigmentation, dermal scattering, dermal blood concentration, blood oxygenation, vessel diameter, and vessel depth. The CIE 1976 color system was used to interpret the resulting spectra as colors. RESULTS: A reduced dermal blood content results in a less red and lighter color. Distribution of a constant volume of blood in smaller vessels results in a redder and darker color. Skin with higher dermal scattering was calculated to be yellower and lighter and skin with increased epidermal pigmentation results in a yellower and darker color. CONCLUSIONS: Redness of PWS skin depends on both the concentration of dermal blood as well as on how it is distributed. PMID- 10455220 TI - A clinical study on the removal of gingival melanin pigmentation with the CO(2) laser. AB - BACKGROUND AND OBJECTIVE: In a previous study, the possibility of removal of dog gingival melanin pigmentation with CO(2) laser therapy was reported. The present study was designed to investigate the effect of the CO(2) laser on human gingival pigmentation and evaluate the clinical outcome. STUDY DESIGN/MATERIALS AND METHODS: A CO(2) laser (output: 6-8 W, pulse duration: 0.2 seconds) was irradiated on the melanin pigmented gingival surface of 10 patients, aged 20-49 years. Follow-up clinical and histopathological evaluations were performed. RESULTS: The CO(2) laser was effective in removing melanin pigmentation in all patients. In the histopathological study, no pigmented-laden cells nor any inflammatory cell infiltration was observed following laser irradiation. No re pigmentation was seen in any case in the first year. However, four of seven cases showed re-pigmentation at 24 months. The re-pigmentation was almost equal to the preoperative state. CONCLUSIONS: The CO(2) laser has proved to be another effective, safe, and easily applicable therapy for the removal of gingival melanin pigmentation. PMID- 10455221 TI - Interstitial laser hyperthermia, a new method in the management of fibroadenoma of the breast: A pilot study. AB - BACKGROUND AND OBJECTIVE: This study attempts to evaluate the effect of interstitial laser hyperthermia in breast fibroadenomas as an outpatient procedure. STUDY DESIGN/MATERIALS AND METHODS: In an uncontrolled prospective study, 27 patients younger than 35 years were subjected to laser phototherapy of their breast fibroadenomas. Under real-time ultrasound monitoring, Nd:YAG laser (1,064 nm wavelength) was used at 2 W for 300 sec (600 J) in a continuous wave mode to produce interstitial hyperthermia. Follow-ups were done at 2, 4, and 8 weeks. Subsequently, excision biopsy of residual lumps was performed. RESULTS: There was significant decrease in clinical and sonographic sizes (P < 0.001). Follow-up ultrasound showed a progressive change of hyperechoic texture, from a heterogeneous to a nearly homogeneous one. There were minimal scars (2-3 mm) and no keloid or abscess formation. CONCLUSION: Interstitial laser hyperthermia is a safe, precise, and minimally invasive outpatient procedure for in situ destruction of breast fibroadenomas. PMID- 10455222 TI - Photoablation of bone by excimer laser radiation. AB - BACKGROUND AND OBJECTIVES: The aim of the present study was to compare the usefulness of mechanical osteotomy tools and common laser systems with the ultraviolet (UV) laser in the field of the central nervous system and its bony capsule. STUDY DESIGN/MATERIALS AND METHODS: The cranial bones of 42 living rats were treated with UV laser radiation with wavelengths of 193 nm and 248 nm. The morphology and physical effects were evaluated by means of optical and scanning electron microscopy. RESULTS: This study shows the special characteristics of excimer versus infrared lasers or mechanical tools, such as high precision, no thermic damage, little depth effect, and no delay of healing processes. CONCLUSION: The excimer laser is an interesting instrument for microsurgery of bones in orthopaedics, neurosurgery, and otolaryngology. PMID- 10455223 TI - Optical phantom materials for near infrared laser photocoagulation studies. AB - BACKGROUND AND OBJECTIVE: Phantoms were developed that simulate tissue with dynamic and static optical properties with which to study the effects of laser irradiation. STUDY DESIGN/MATERIALS AND METHODS: Albumen, agar, and an absorbing dye (Naphthol Green) were combined to form a phantom with heat sensitive optical properties to mimic tissue response. The optical properties of this phantom were measured by using the added absorber technique. A polyacrylamide phantom with static optical properties was designed with the equivalent values of micro(a) and micro'(s) by combining appropriate concentrations of Naphthol Green and Intralipid-10%. RESULTS: The absorption and reduced scattering coefficient of the phantoms were 0. 50 +/- 0.04 cm(-1) and 2.67 +/- 0.07 cm(-1) respectively, in the native state at 805 nm. In the coagulated state, the absorption and scattering coefficient were 0.7 +/- 0.1 cm(-1) and 13.1 +/- 0.5 cm(-1) respectively. CONCLUSION: Two phantoms with dynamic or static optical properties were developed with properties similar to tissue. They may be used in future studies of opto thermal effects in tissues. PMID- 10455224 TI - Treatment of venous malformations with an intense pulsed light source (IPLS) technology: A retrospective study. AB - BACKGROUND AND OBJECTIVE: The intense pulsed light source (IPLS) technology provides an innovative concept in the treatment of vascular lesions. We investigated the effectiveness of IPLS in the treatment of venous malformations. STUDY DESIGN/MATERIALS AND METHODS: A retrospective study of 11 patients with venous malformations (VMA) treated with IPLS was initiated. Clinical VMA characteristics recorded were size and location. Data collected included treatment parameters (filters, pulse duration, fluence, and pulse sequencing), % clearance, and side effects (e.g., swelling, blisters, crusting, pain, altered pigmentation, and scarring). RESULTS: Good and very good (70-100%) clearance was achieved in 8 malformations smaller than 100 cm(2). Especially small lesions needed only 2-3 treatments. Three VMA larger than 100 cm(2) could be cleared well in an average of 18 sessions. The most frequently used parameters were the 590 nm filter in long pulse mode, triple pulses, and fluences at an average of 80.4 J/cm(2). Side effects included prolonged erythema in 23.6, swelling in 17.9%, crusting in 4.7%. Bleeding, hypo-, hyperpigmentation, and scarring were rare (0.9% respectively). CONCLUSION: IPLS presents an effective method for treating VMA, especially small malformations, with a justifiable rate of side effects when optimal parameters are chosen. PMID- 10455225 TI - Comparison of flash lamp pulsed-dye laser (585 nm) and conventional surgery in the delay of random dorsal rat flaps. AB - BACKGROUND AND OBJECTIVE: Delay is a basic surgical technique used by flap surgeons to improve the blood supply to the distal parts of a random skin flap. The aim of this study was to determine whether a scarless delay can be done by the use of the flash lamp pulsed-dye laser operating at a wavelength of 585 nm. STUDY DESIGN/MATERIALS AND METHODS: The pilot study showed that 6 J/cm(2) had a selective photothermolysis effect and therefore was chosen for testing the delay procedure on 15 rats. The percentage of flap necrosis of this group was compared to the results of 15 rats that underwent delay by surgery and 15 rats that were not treated prior to flap surgery (control group). RESULTS: Laser delay of McFarlane flaps resulted in an average of 15.5% smaller necrotic area compared to the control group (52.7% +/- 14.4% and 68.2% +/- 9.6%, respectively, P < 0.01) and was as effective as surgical delay (53.3% +/- 13.6%). CONCLUSIONS: The results indicate that the flash lamp pulsed-dye laser operating at 585 nm is effective for delaying cutaneous flaps in the rat model. PMID- 10455226 TI - Disruption of the cytochrome c gene in xylose-utilizing yeast Pichia stipitis leads to higher ethanol production. AB - The xylose-utilizing yeast, Pichia stipitis, has a complex respiratory system that contains cytochrome and non-cytochrome alternative electron transport chains in its mitochondria. To gain primary insights into the alternative respiratory pathway, a cytochrome c gene (PsCYC1, Accession No. AF030426) was cloned from wild-type P. stipitis CBS 6054 by cross-hybridization to CYC1 from Saccharomyces cerevisiae. The 333 bp open reading frame of PsCYC1 showed 74% and 69% identity to ScCYC1 and ScCYC7, respectively, at the DNA level. Disruption of PsCYC1 resulted in a mutant that uses the salicylhydroxamic acid (SHAM)-sensitive respiratory pathway for aerobic energy production. Cytochrome spectra revealed that cytochromes c and a.a(3) both disappeared in the cyc1-Delta mutant, so no electron flow through the cytochrome c oxidase was possible. The cyc1-Delta mutant showed 50% lower growth rates than the parent when grown on fermentable sugars. The cyc1-Delta mutant was also found to be unable to grow on glycerol. Interestingly, the mutant produced 0.46 g/g ethanol from 8% xylose, which was 21% higher in yield than the parental strain (0.38 g/g). These results suggested that the alternative pathway might play an important role in supporting xylose conversion to ethanol under oxygen-limiting conditions. PMID- 10455227 TI - Total number of coding open reading frames in the yeast genome. AB - At the end of 1996 we approximated the total number of protein coding ORFs in the Saccharomyces cerevisiae genome, based on their properties, as 4700-4800. The number is much smaller than the 5800 which is widely accepted. According to our calculations, there remain about 200-300 orphans-ORFs without known function or homology to already discovered genes, which is only about 5% of the total number of genes. Our results would be questionable if the analysed set of known genes was not a statistically representative sample of the whole set of protein coding genes in the S. cerevisiae genome. Therefore, we repeated our estimation using recently updated databases. In the course of the last 18 months, previously unknown functions of about 500 genes have been found. We have used these to check our method, former results and conclusions. Our previous estimation of the total number of coding ORFs was confirmed. PMID- 10455228 TI - Analysis of the peroxisomal acyl-CoA oxidase gene product from Pichia pastoris and determination of its targeting signal. AB - Acyl-CoA oxidase (Pox1p) is involved in the beta-oxidation of fatty acids and is targeted to the peroxisomal matrix via the use of different signals in various organisms. In rat, mouse and human, Pox1p contains a canonical peroxisomal targeting signal 1 (PTS1), whereas in the yeasts Candida tropicalis, Saccharomyces cerevisiae, C. maltosa and Yarrowia lipolytica neither a PTS1 nor a PTS2 sequence is present, suggesting that Pox1p might be targeted to the peroxisomes via a third unknown pathway. Alternatively, since proteins lacking a PTS sequence can enter peroxisomes in association with other polypeptides containing a PTS, Pox1p might 'piggy-back' its way into the peroxisomal matrix together with other proteins. To understand the mechanism of peroxisomal targeting of a yeast Pox1p, we cloned the Pichia pastoris POX1 gene to study the pathway of import of PpPox1p into peroxisomes. The gene was cloned by PCR, hybridization and plasmid rescue. The 2157 bp gene encodes a protein with a predicted molecular weight of 80 kDa. Antisera against PpPox1p detected a protein specifically induced on oleate with an apparent molecular weight of 72 kDa. Immunolocalization studies confirmed the peroxisomal localization of PpPox1p. The carboxy-terminus of PpPox1p ends with a PTS1-like sequence, APKI. The sequence PKI was necessary for transport of PpPox1p into peroxisomes and interacted with the PTS1 receptor, Pex5p. Furthermore, addition of the sequence APKI to the C terminus of the green fluorescent protein directed this fusion protein to the peroxisome. Therefore, PpPox1p uses the PTS1 pathway for its import into peroxisomes. PMID- 10455229 TI - Transcription of the HXT4 gene is regulated by Gcr1p and Gcr2p in the yeast S. cerevisiae. AB - Glucose transport and glycolysis are two sequential events which are regulated by both physiological and environmental signals in the yeast Saccharomyces cerevisiae. Transcription of the HXT4 gene was found to be regulated by Gcr1p and Gcr2p, transcription factors that are required for the regulated high level transcriptions of glycolytic genes. Transcription of HXT4 decreased about 35-fold in gcr1 mutant and two-fold in gcr2 mutant yeast cells. However, transcription of other HXT genes was not affected at a significant level by gcr1 or gcr2 mutations. Overproduction of Gcr1p from an inducible promoter resulted in a 15 64% increase in transcription of HXT4, depending on the growth conditions. Gel mobility shift assays performed with the purified DNA binding domain of Gcr1p and the UAS region of the HXT4 gene showed that Gcr1p interacts directly with multiple sites on the HXT4 UAS region. These results indicate that Gcr1p and Gcr2p coordinate the transcription of HXT4 and glycolytic genes. PMID- 10455230 TI - Hansenula polymorpha Pex1p and Pex6p are peroxisome-associated AAA proteins that functionally and physically interact. AB - We have cloned the Hansenula polymorpha PEX1 and PEX6 genes by functional complementation of the corresponding peroxisome-deficient (pex) mutants. The gene products, HpPex1p and HpPex6p, are ATPases which both belong to the AAA protein family. Cells deleted for either gene (Deltapex1 or Deltapex6) were characterized by the presence of small peroxisomal remnants which contained peroxisomal membrane proteins and minor amounts of matrix proteins. The bulk of the matrix proteins, however, resided in the cytosol. In cell fractionation studies HpPex1p and HpPex6p co-sedimented with the peroxisomal membrane protein HpPex3p in both wild-type cells and in Deltapex4, Deltapex8 or Deltapex14 cells. Both proteins are loosely membrane-bound and face the cytosol. Furthermore, HpPex1p and HpPex6p physically and functionally interact in vivo. Overexpression of PEX6 resulted in defects in peroxisomal matrix protein import. By contrast, overexpression of PEX1 was not detrimental to the cells. Interestingly, co-overproduction of HpPex1p rescued the protein import defect caused by HpPex6p overproduction. Overproduced HpPex1p and HpPex6p remained predominantly membrane-bound, but only partially co localized with the peroxisomal membrane protein HpPex3p. Our data indicate that HpPex1p and HpPex6p function in a protein complex associated with the peroxisomal membrane and that overproduced, mislocalized HpPex6p prevents HpPex1p from reaching its site of activity. PMID- 10455231 TI - Distribution of the actin cytoskeleton during the cell cycle of Yarrowia lipolytica and the visualization of the tubulin cytoskeleton by immunofluorescence. AB - Actin distribution was examined during the cell cycle of the dimorphic yeast Yarrowia lipolytica, showing the correlation between bud growth, nuclear migration and rearrangement of the actin cytoskeleton. The results correspond with observations made in cells of Saccharomyces cerevisiae, S. uvarum and Candida albicans. Localization of actin was also determined in hyphal cells, where actin is stained predominantly in the tip and also at the septum of hyphae. The standard methods used for tubulin immunostaining in S. cerevisiae and C. albicans cells were adapted for application in Y. lipolytica. PMID- 10455232 TI - High-yield secretion of recombinant gelatins by Pichia pastoris. AB - Recombinant non-hydroxylated gelatins based on mouse type I and rat type III collagen sequences were secreted from the methylotrophic yeast Pichia pastoris, using the Saccharomyces cerevisiae alpha-mating factor prepro signal. Proteolytic degradation could be minimized to a large extent by performing fermentations at pH 3.0 and by adding casamino acids to the medium, even though gelatin is extremely susceptible to proteolysis due to its open, unfolded structure. Proteolytic cleavage at specific mono-arginylic sites, by a putative Kex2-like protease, could be successfully abolished by site-directed mutagenesis of these sites. Production levels as high as 14.8 g/l clarified both were obtained, using multicopy tranformants. To our knowledge, this represents the highest level of heterologous protein secretion reported to date for P. pastoris. PMID- 10455233 TI - Deletion of SFI1, a novel suppressor of partial Ras-cAMP pathway deficiency in the yeast Saccharomyces cerevisiae, causes G(2) arrest. AB - When glucose is added to Saccharomyces cerevisiae cells grown into stationary phase or on non-fermentable carbon sources a rapid loss of heat stress resistance occurs. Mutants that retain high stress resistance after addition of glucose are called 'fil', for deficient in fermentation induced loss of stress resistance. Transformation of the fil1 mutant, which harbours a point mutation in adenylate cyclase, with a yeast gene library on a single copy plasmid resulted in transformants that were again stress-sensitive. One of the genes isolated in this way was a gene of previously unknown function. We have called it SFI1, for suppressor of fil1. SFI1 is an essential gene. Combination of Sfi1 and cAMP pathway mutations indicates that Sfi1 itself is not involved in the cAMP pathway. Conditional sfi1 mutants did not show enhanced heat resistance under the restrictive condition, whereas overexpression of SFI1 rendered cells heat sensitive. Sfi1 may be a downstream target of the protein kinase A pathway, but its precise relationship with heat resistance remains unclear. Further analysis showed that Sfi1 is required for cell cycle progression, more specifically for progression through G(2)-M transition. Cells expressing SFI1 under the control of a galactose-inducible promoter arrest after addition of glucose as doublets of undivided mother and daughter cells. These doublets contain a single nucleus and lack mitotic spindles. Sfi1 shares homology with Xenopus laevis XCAP-C, a protein required for chromosome assembly. The conserved residues between these two proteins show a strong bias for charged amino acids. Hence, Sfi1 might be required for correct mitotic spindle assembly and its precise role might be in chromosome condensation. In conclusion, we have identified an essential function in the G(2)-M transition of the cell cycle for a yeast gene of previously unknown function. PMID- 10455234 TI - Mutations in the Saccharomyces cerevisiae gene SAC1 cause multiple drug sensitivity. AB - Wild-type yeast Saccharomyces cerevisiae are surprisingly resistant to a wide range of drugs and agents. We had previously isolated novobiocin-sensitive mutants to aid the study of the intracellular target for this drug. Characterization of one of these mutants, mds1, revealed that it was sensitive not only to novobiocin but also to a wide range of drugs. The nature of this multiple drug-sensitive phenotype was shown to be different from that of previously isolated multiple drug-sensitive mutants. We have shown that the multiple drug-sensitivity of mds1 is due to mutations within the gene SAC1 and have identified a variety of mutations within the gene from the Mds1 strain. SAC1 encodes a protein which has been previously implicated in the correct function of the actin cytoskeleton, in inositol metabolism, in ATP transport in the endoplasmic reticulum and in Sec14p (PI-TP) function. We have shown that multiple drug-sensitivity is a new phenotype seen in some, but not all, of the previously characterized sac1 mutants. Based on our findings, we propose a mechanism by which Sac1p could affect drug resistance and also mediate other effects on cell growth. PMID- 10455235 TI - Schizosaccharomyces pombe homologue of glutathione peroxidase, which does not contain selenocysteine, is induced by several stresses and works as an antioxidant. AB - We have cloned a gene of Schizosaccharomyces pombe homologues to the glutathione peroxidase gene. The cloned gene, named gpx1(+), encoded a protein that was 158 amino acids in length and had a molecular mass of 18 kDa. The gpx1(+) gene is homologous with many glutathione peroxidase genes but the selenocysteine codon (UGA) position of mammalian genes is a cysteine codon (UGU) in S. pombe. gpx1(+) mRNA was induced by various stresses, including oxidative stress, osmostress and heat stress. These stresses activate the Wis1-Sty1/Spc1 MAP kinase cascade in S. pombe. Transcriptional factors Atf1 and Pap1 are under the control of this MAP kinase. In the disruption of the atf1(+) gene, gpx1(+) was not transcribed or induced. However, the expression of gpx1(+) was not affected by the disruption of the pap1(+) gene. These results indicated that gpx1(+) was under the control of transcription factor Atf1. Catalase can detoxicate H(2)O(2) in the same way as GPx and the disruptant of the catalase gene of S. pombe is hypersensitive to H(2)O(2). The catalase gene disruptant of S. pombe harbouring multicopy plasmid containing gpx1(+) restored the hypersensitivity to H(2)O(2) of the catalase gene disruptant. These results suggest that Gpx1 acts as a scavenger of H(2)O(2) in vivo. PMID- 10455236 TI - Molecular cloning of homologs of RAS and RHO1 genes from Cryptococcus neoformans. AB - We cloned and sequenced homologs of RAS(CnRAS) and RHO1(CnRHO1) genes from Cryptococcus neoformans. The proteins encoded by the CnRAS and CnRHO1 genes contained 216 and 197 amino acids, respectively. The deduced amino acid sequence of the CnRAS gene shared a high degree of sequence identity with the Ras proteins in other fungal species: Coprinus cinereus(76%), Lentinula edodes(74%), Saccharomyces cerevisiae RAS2(72%), and Schizosaccharomyces pombe(68%). The deduced amino acid sequence of the CnRHO1 gene shared a high degree of sequence identity with the Rho1 proteins in other fungal species: Candida albicans(78%), S. pombe(77%) and S. cerevisiae(76%). The deduced proteins contained GTP-binding and GTP-hydrolysis domains, and the prenylation site that are conserved among the small G protein superfamily. The synthetic peptides that contained the C-terminal amino acid sequence of the CnRas and CnRho1 proteins were geranylgeranylated. PMID- 10455237 TI - Designer deletion and prototrophic strains derived from Saccharomyces cerevisiae strain W303-1a. AB - We report the construction of Saccharomyces cerevisiae strains isogenic to W303 1a that are designed to allow efficient genetic analysis. To facilitate the generation of null alleles of target genes by PCR-mediated gene disruption, we constructed designer deletion alleles of the ARG4, TRP1 and URA3 genes. In addition, a single pair of oligonucleotide primers were designed that can be used to amplify any of several marker genes for use in PCR-mediated gene disruption. A new version of the 'reusable' hisG-URA3-hisG cassette was constructed for use in PCR-mediated gene disruption. Finally, to facilitate the formation of isogenic diploids by selection, we constructed strains that contain combinations of wild type alleles of ADE2, HIS3, LEU2, TRP1 and URA3. PMID- 10455238 TI - Characterization of ionized heterocyclic carbenes by ion-molecule reactions. AB - 1,2-Hydrogen shift isomers of ionized pyridine, thiazole and imidazole are readily characterized by the study of their associative ion-molecule reactions with dimethyl disulfide in the quadrupole collision cell of a new hybrid sector quadrupole-sector mass spectrometer. Efficient trapping reactions of CH(3)S(.) radicals are indeed observed and the actual structure of the adduct [M + CH(3)S](+) ions is clearly indicated by their high-energy collisional activation mass spectra. These trapping reactions are not observed for the 'conventional' pyridine, thiazole and imidazole molecular ions, which only react by charge exchange producing m/z 94, [CH(3)SSCH(3)](*+), ions. PMID- 10455239 TI - Comparison between collision induced dissociation of electrosprayed protonated peptides in the up-front source region and in a low-energy collision cell. AB - A systematic comparison between the up-front Collision induced dissociation (CID) mass spectra and low-energy CID tandem mass spectra from twenty-one singly and/or doubly charged peptides has been made. CID spectra of the peptides were recorded at different electrode voltages in the up-front source region of a single quadrupole instrument and different collision energies in the collision cell of a tandem quadrupole instrument. It was observed that up-front CID and low-energy CID yielded comparable product ion spectra from protonated peptides, and that the instrumental settings necessary for obtaining comparable CID mass spectra from the two methods are correlated. PMID- 10455240 TI - Simplified sample preparation for the analysis of oligonucleotides by matrix assisted laser desorption/ionisation time-of-flight mass spectrometry. AB - Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS) for the analyses of oligonucleotides has generally been carried out using negative ionisation conditions, usually following ammonium ion-exchange chromatography and the addition of ammonium buffers to the MALDI matrix. The molecular ion region is complex, due to the varying degrees of ammoniation of the phosphate backbone of the oligonucleotide. This gives rise to an overall decrease in sensitivity compared with similar size peptides and can cause ambiguity of assignment of the relative molecular mass of the sample. This study describes the use of H(+) ion exchange resin in situ as the means of removing alkali metal ions from the phosphate backbone of the oligonucleotide. An increase in resolution, sensitivity and identification of the molecular species is reported, with little or no difference in sensitivity observed between positive or negative ionisation spectra. This method is now used for routine screening of synthetic oligonucleotides with a gain in sensitivity of 1-2 orders of magnitude compared with previous methods, and mass assignment errors of +/-0.1% are routinely recorded for externally calibrated data. PMID- 10455241 TI - The citropin peptides from the skin glands of the Australian Blue Mountains tree frog Litoria citropa. Part 2: sequence determination using electrospray mass spectrometry. AB - A combination of electrospray mass spectrometry, Lys-C digest/mass spectrometry and automated Edman sequencing provides the amino acid sequences of nineteen citropin peptides isolated from the granular dorsal and submental glands of the Blue Mountains tree frog Litoria citropa. Citropin 1.1 [Gly Leu Phe Asp Val Ile Lys Lys Val Ala Ser Val Ile Gly Gly Leu (NH(2))] and citropin 1.2 [Gly Leu Phe Asp Ile Ile Lys Lys Val Ala Ser Val Val Gly Gly Leu (NH(2))] are the two major skin peptides: both show significant wide-spectrum antibacterial activity. PMID- 10455242 TI - Quantification of breath isoprene using the selected ion flow tube mass spectrometric analytical method. AB - We have used our selected ion flow tube mass spectrometric method (SIFT-MS) to study isoprene levels in the alveolar breath of 29 healthy volunteers during normal working hours at the varying states of nutrition occurring during this period. Quantification of the breath isoprene was achieved using O(2)(+) precursor ions to avoid complications which can arise when using H(3)O(+) precursor ions for isoprene analysis. The present data indicate that the spread of the alveolar isoprene levels in this sample of healthy individuals is 22 to 234 ppb and that the mean value is 83 ppb with a standard deviation of 45 ppb. These levels are compared with those previously determined using other techniques and are seen to be at the low end of the values previously reported. The present studies are a prelude to an investigation of the proposed correlation of breath isoprene levels with psychological, physical and biochemical stress. PMID- 10455243 TI - Analysis of negatively 'charge tagged' DNA by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - An improvement in detectability and stability of DNA analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) using oligonucleotides modified with a neutralized backbone and a fixed single, positive charge was recently reported. The attachment of the positive charge requires a primary amino group, limiting this approach to accordingly functionalized DNA. The method described here uses backbone neutralized DNA for the same purpose, with a single unmodified phosphate in the DNA backbone carrying the negative charge. Thus, no chemical modification other than neutralizing the remaining charges on the phosphorothioates is required. This is performed in a single methylation step. The enhancement in sensitivity is comparable to that for DNA carrying a single positive charge, interestingly even when using the same non protonating matrix. The mechanistic implications of these findings regarding the MALDI process are discussed. The DNA derivatization methods presented help to make MALDI-MS of DNA applicable and competitive for genome analysis and medical diagnostics. PMID- 10455244 TI - Rapid analysis of tetracycline antibiotics by combined solid phase microextraction/high performance liquid chromatography/mass spectrometry. AB - The technique of solid phase microextraction (SPME) combined on-line with high performance liquid chromatography/mass spectrometry (HPLC/MS) has been applied to the analysis of seven tetracycline analogues. Rapid baseline separation was achieved in under 5 min using a short 3 microm RP-18e cartridge column. Optimisation of the SPME procedure is described including choice of extracting fibre and modification of the sample by heating or salting out of the analytes. Detection limits of 4-40 ng/mL were obtained for the various analogues from extracted aqueous samples and absolute amounts of analyte extracted by the method determined using external calibration. To demonstrate the applicability of the technique for real samples the extraction of tetracycline from milk is described. PMID- 10455245 TI - Principal components scores and loadings plots for visualisation of the electrospray ionisation liquid chromatography mass spectra of a mixture of chlorophyll degradation products at different cone voltages. AB - Positive ion electrospray liquid chromatography mass spectrometry was performed on a mixture of allomers obtained from the degradation of chlorophyll a, at three cone voltages. An approach is described for obtaining principal components loadings and scores plots, involving mass selection, normalisation and standardisation of the data, principal components analysis and three dimensional projections. The loadings plots group ions which are assigned to five major compounds in the mixture by reference to the scores. At higher cone voltage fragmentation and differentiation between compounds with identical molecular weights is observed. PMID- 10455246 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of cod liver oil and the effect of analyte/matrix concentration on signal intensities. AB - Cod liver oil (CLO) is known to contain a complex mixture of triacylglycerols (TAGs) in which the component fatty acids include: myristic (C(14:0), M), C(14:1) (M(1)), palmitic (C(16:0), P), palmitoleic (C(16:1), P(1)), stearic (C(18:0), S), oleic (C(18:1), O), linoleic (C(18:2), L), arachidic (C(20:0), A), C(20:1) (A(1)), eicosapentaenoic (EPA, C(20:5), A(5)), docosanoic (C(22:0), D), docosaenoic (C(22:1), D(1)), and docosahexaenoic (DHA, C(22:6), D(6)). Because of the presence of EPA and DHA in cod liver oil, it has been used for several generations as a nutritional supplement, and recommended for the relief of various physiological ailments including arthritis, depression, and high blood pressure. Consequently, it was of interest to develop a sample preparation protocol that would enable rapid screening of such a chemically complex and nutritionally useful oil. Thus, we have analyzed two commercial brands of cod liver oil by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). There was no significant difference between the mass spectral profile of the two CLO brands. alpha-Cyano-4-hydroxycinnamic acid, dissolved in acetonitrile/tetrahydrofuran, was used as the matrix. MALDI-TOFMS produced only sodiated triacylyglycerol molecules [M + Na](+). Based on the sodiated TAGs, 64 TAG assignments were made, and these include MM(1)L, MML, MMO and MMS, M(1)P(1)L MP(1)L, P(1)P(1)P, PPP, P(1)P(1)Ln, P(1)PLn, PPL, PPO, P(1)LnLn, PLnLN, PLLn, PLL, POL, POO, P(1)A(6)Ln, P(1)A(5)Ln, P(1)A(5)L, PA(5)L PA(5)O, PP(1)D(6), OOL, OOO, SOO, SSS, P(1)LnD(6), PLnD(6), PLD(6), POD(6) (or P(1)A(5)A(1)), PA(5)A(1), OLA, OLA(1), SLA(1), SOA(1), SSA, LA(5)A(5) (or P(1)A(5)D(6)), OA(5)A(5) (or PA(5)D(6)), SA(5)A(5), LnA(1)A(5), OOD(6), SOD(6), SSD(6), LA(1)D(6), OA(1)D(6), OA(5)D(6), SA(5)D(6), SA(5)D(5), D(6)A(1)O, D(6)A(1)S, D(1)A(1)O, DA(1)O, D(1)D(6)O, and DD(6)O. The sample preparation method developed in this study could be used for the routine screening of oils that contain similar types of polyunsaturated TAGs. PMID- 10455247 TI - A simple method for characterizing a neutral species lost in a metastable decomposition using a tandem mass spectrum of a naturally abundant isotopic ion containing a (13)C atom. PMID- 10455248 TI - Antagonism of the mGlu5 agonist 2-chloro-5-hydroxyphenylglycine by the novel selective mGlu5 antagonist 6-methyl-2-(phenylethynyl)-pyridine (MPEP) in the thalamus. AB - Our previous work has shown that Group I mGlu receptors participate in thalamic sensory processing in vivo. However, unequivocal demonstration of mGlu5 participation has not been possible due to the lack of specific ligands. We have therefore made a preliminary study of the in vivo actions of the agonist (R,S)-2 Chloro-5-hydroxyphenylglycine [CHPG] and the novel mGlu5 antagonist 6-methyl-2 (phenylethynyl)-pyridine [MPEP] in order to characterize their suitability for functional studies. Iontophoretically administered MPEP selectively antagonized excitatory responses of single rat thalamic neurones to CHPG compared to the broad-spectrum mGlu agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylate. In contrast, the established mGlu1 and mGlu5 antagonist (S)-4-carboxyphenylglycine reduced responses to both agonists. These findings are the first demonstration of an in vivo action of CHPG and its antagonism by a selective mGlu5 antagonist. Furthermore MPEP appears to be a good tool for functional studies of mGlu5. PMID- 10455249 TI - Rundown of somatodendritic N-methyl-D-aspartate (NMDA) receptor channels in rat hippocampal neurones: evidence for a role of the small GTPase RhoA. AB - Actin filament (F-actin) depolymerization leads to the use-dependent rundown of N methyl-D-aspartate (NMDA) receptor activity in rat hippocampal neurones. Depolymerization is promoted by Ca2+ which enters the cells via NMDA receptor channels. The ras homologue (Rho) GTPases (RhoA, Rac1 and Cdc42) promote actin polymerization and thus control the actin cytoskeleton. We have investigated, by means of the whole-cell patch clamp technique, whether the actin fibres which interact with NMDA receptors are controlled by Rho GTPases. In the presence of intracellular ATP which attenuates rundown, the C3 toxin from Clostridium (C.) botulinum was used to inactivate RhoA. Indeed, it enhanced the use-dependent rundown of NMDA-evoked inward currents to a level similar to that obtained in the absence of ATP. Lethal toxin from Clostridium sordellii which inactivates Rac1 and Cdc42 lacked this effect. We suggest that the function of somatodendritic NMDA receptor channels in rat hippocampal neurones can be modulated by RhoA via its action on F-actin. PMID- 10455250 TI - The effect of deep pore mutations on the action of phenylalkylamines on the Kv1.3 potassium channel. AB - We investigated the action of the phenylalkylamines verapamil and N-methyl verapamil on the Kv1.3 potassium channel using the whole-cell configuration of the patch-clamp technique. Our goal was to identify their binding as a prerequisite for using the phenylalkylamines as small, well-defined molecular probes, not only to expand the structural findings made with peptide toxins or by crystallization, but also to use them as lead compounds for the generation of more potent and therefore more specific K+ channel modulators. Competition experiments with charybdotoxin, known to interact with external residues of Kv1.3, showed no interaction with verapamil. The internal application of quarternary N-methyl-verapamil in combination with verapamil suggested competition for the same internal binding site. Verapamil affinity was decreased 6 fold by a mutation (M395V) in a region of the internal pore which forms part of the internal tetraethylammonium (TEA+) binding site, although mutations at neighbouring residues (T396 and T397) were without effect. Modification of C-type inactivation by mutations in the internal pore suggest that this region participates in the inactivation process. The action of phenylalkylamines and local anaesthetics on L-type Ca2+ channels and Na channels, respectively, and verapamil on Kv1.3 indicate very similar blocking mechanisms. This might allow the use of these compounds as molecular probes to map the internal vestibule of all three channel types. PMID- 10455251 TI - RS-127445: a selective, high affinity, orally bioavailable 5-HT2B receptor antagonist. AB - Efforts to define precisely the role of 5-HT2B receptors in normal and disease processes have been hindered by the absence of selective antagonists. To address this deficiency, we developed a series of naphthylpyrimidines as potentially useful 5-HT2B receptor antagonists. RS-127445 (2-amino-4-(4-fluoronaphth-1-yl)-6 isopropylpyrimidine) was found to have nanomolar affinity for the 5-HT2B receptor (pKi = 9.5+/-0.1) and 1,000 fold selectivity for this receptor as compared to numerous other receptor and ion channel binding sites. In cells expressing human recombinant 5-HT2B receptors, RS-127445 potently antagonized 5-HT-evoked formation of inositol phosphates (pK(B) = 9.5+/-0.1) and 5-HT-evoked increases in intracellular calcium (pIC50 = 10.4+/-0.1). RS-127445 also blocked 5-HT-evoked contraction of rat isolated stomach fundus (pA2 = 9.5+/-1.1) and (+/-)alpha methyl-5-HT-mediated relaxation of the rat jugular vein (pA2 = 9.9+/-0.3). RS 127445 had no detectable intrinsic activity in these assays. In rats, the fraction of RS-127445 that was bioavailable via the oral or intraperitoneal routes was 14 and 60% respectively. Intraperitoneal administration of RS-127445 (5 mg kg(-1)) produced plasma concentrations predicted to fully saturate accessible 5-HT2B receptors for at least 4 h. In conclusion, RS-127445 is a selective, high affinity 5-HT2B receptor antagonist suitable for use is vivo. The therapeutic potential of this molecule is being further evaluated. PMID- 10455252 TI - Characterization of the inflammatory response to proteinase-activated receptor-2 (PAR2)-activating peptides in the rat paw. AB - In the present study, we have observed the development of an inflammatory reaction in the rat hindpaw, following the injection of specific agonists of PAR2 (two PAR2 activating peptides). This inflammation was characterized by oedema and granulocyte infiltration. Two selective PAR2 activating peptides, SLGRL-NH2 and trans-cinnamoyl-LIGRLO-NH2 induced significant oedema in the rat hindpaw from 1-6 h following subplantar injection. Six hours after the PAR2-activating peptide injection, the paw tissues showed a complete disruption of tissue architecture along with an inflammatory cell infiltrate. In the inflamed paw, PAR2 immunoreactivity was expressed on endothelial cells as well as on the infiltrating inflammatory cells. The oedema induced by the injection of the two PAR2 activating peptides was slightly reduced in rats pre-treated with compound 48/80, but was not modified by pre-treatment of rats with cromolyn, a mast cell stabilizer. Pre-treatment of rats with a cyclo-oxygenase inhibitor (indomethacin) or a nitric oxide synthase inhibitor (L-N(omega)-nitro-L-arginine methyl ester) had no effect on the oedema induced by the PAR2-activating peptides. These results demonstrate that the administration of PAR2-activating peptides into the rat paw induced an acute inflammatory response characterized by a persistent oedema (at least 6 h) and granulocyte infiltration. The PAR2-induced inflammatory response occurred through a mechanism largely independent of mast cell activation, and of the production of prostanoids and nitric oxide. PMID- 10455253 TI - Pharmacological characterization of endothelin receptor subtypes in the guinea pig prostate gland. AB - Experiments have been conducted to investigate the actions of endothelins on the guinea-pig prostate gland. Saturation experiments with [125I]-endothelin-1 (2-800 pM) in guinea-pig prostatic homogenates indicated the presence of high affinity binding sites with an equilibrium dissociation constant (KD) of 230+/-50 pM, a maximum number of binding sites (Bmax) of 52+/-16 fmol mg(-1) protein or 269+/-61 fmol g(-1) tissue and a Hill coefficient (nH) of 1.01+/-0.03 (n = 3). Competition experiments revealed that binding of [125I]-endothelin-1 (20 pM) was inhibited with the following order of potency: endothelin-1 >>BQ-788 (N-cis-2,6 dimethylpiperidinocarbonyl-L-gamma-methyl-Leu-D-Trp[1-+ ++CO2CH3-D-Nle-ONa])> BQ 123 (cyclo-D-Asp-L-Pro-D-Val-Leu-D-Trp) > or = sarafotoxin S6c. At concentrations with negligible influence on smooth muscle tone, endothelin-1, endothelin-2 and sarafotoxin S6b (1 nM-0.1 microM) produced concentration-dependent potentiation of the contractions evoked by electrical field stimulation with trains of 20 pulses at 10 Hz every 50 s, 0.5 ms pulse width and a dial setting of 60 V. In contrast, the endothelin ET(B) receptor-preferring agonist endothelin-3 (1 nM- 1 microM) was much less potent, and the endothelin ET(B) receptor-selective agonists sarafotoxin S6c and BQ-3020 (Ac-[Ala11,15]-endothelin-1 (6-21)), up to 1 microM, were without effect. The endothelin ET(A) receptor antagonist BQ-123 (1 microM) markedly inhibited the potentiation induced by endothelin-1, endothelin-2 and sarafotoxin S6b while the endothelin ET(B) receptor antagonist BQ-788 (1 microM) was less effective. While our binding data indicates the presence of ET(A) and ET(B) binding sites in the guinea-pig prostate, the endothelin-induced facilitation of neurotransmission to the prostatic smooth muscle is mediated largely via activation of endothelin receptors of the ET(A) subtype. PMID- 10455254 TI - Treatment with inverse agonists enhances baseline atrial contractility in transgenic mice with chronic beta2-adrenoceptor activation. AB - In this study, we investigate whether chronic treatment with beta-adrenoceptor (betaAR) ligands with inverse agonist activity enhances myocardial beta2AR mediated atrial tension more than neutral antagonists in transgenic mice (TG35). These mice exhibit chronic adrenoceptor activation because they possess a greater number of constitutively active receptors than wild type mice due to cardiac specific overexpression of human betaARs. TG35 and wild type mice were chronically treated for 90 h with three inverse agonists, ICI-118,551, propranolol, and carvedilol, and one neutral antagonist, alprenolol. After 96 h, we compared the basal and isoprenaline-stimulated (10 microM) increase in atrial tension in treated or untreated TG35 mice and wild type mice. In parallel, to determine the effect of chronic betaAR ligand treatment on the amounts of G protein receptor kinase-2 (GRK-2) and G proteins, we performed Western blotting on myocardial cytosolic and membrane proteins. Atria from the TG35 mice treated with inverse agonists showed increases in the baseline tension compared to those from alprenolol/vehicle-treated mice. ICI-118,551 and propranolol treatment restored the elevated myocardial G-inhibitory protein (Gialpha) levels to that of wild type. Also, treatment with inverse agonists upregulated G-stimulatory protein (Gsalpha) levels and GRK2 above those levels in vehicle-treated TG35 or wild type mice. The increased baseline atrial tension was reversed by the addition of ICI-118,551. Overall, our data suggests that inverse agonists enhance baseline atrial tension more than neutral antagonists. Based on this, we propose that upregulation of the active conformation of the beta2ARs, Gsalpha protein and restoration of Gialpha as three possible mechanisms to explain this enhanced receptor activity. Therefore, the favourable effects of some ligands used in pathological conditions involving chronic adrenoceptor activation may be due to the inverse agonist activity of the ligand. PMID- 10455255 TI - Characterization of the [125I]-neurokinin A binding site in the circular muscle of human colon. AB - Neurokinin A (NKA) is a potent contractile agonist of human colon circular muscle. These responses are mediated predominantly through tachykinin NK2 receptors. In the present study, the NK2 receptor radioligand [125I]-NKA has been used to characterize binding sites in this tissue, using tachykinin agonists and antagonists. 125INKA labelled a single, high affinity binding site. Specific binding (95% of total binding) of [125I]-NKA was saturable (K(D) 0.47+/-0.05 nM), of high capacity (Bmax 2.1+/-0.1 fmol mg(-1) wet weight tissue) and reversible (kinetically derived K(D) 0.36+/-0.07 nM). The rank order of agonists competing for the [125I]-NKA binding site was neuropeptide gamma (NPgamma) > or = NKA > or = [Lys5, MeLeu9,Nle10]NKA (4-10) (NK2 agonist) >> substance P (SP) > neurokinin B (NKB) > or = [Pro9]SP (NK1 agonist) >> senktide (NK3 agonist), indicating binding to an NK2 site. The nonpeptide selective NK2 antagonist SR48968 showed higher affinity for the [125I]-NKA site than selective peptide NK2 antagonists. The rank order of potency for NK2 antagonists was SR48968 > or = MEN11420 > GR94800 > or = MEN10627 > MEN10376 > or = R396. The NK1 antagonist SR140333 was a weak competitor. The competition curve for SP could be resolved into two sites. When experiments were repeated in the presence of SR140333 (0.1 microM), the curve for SP became monophasic and showed a significant shift to the right, whereas curves to NKA and NKB were unaffected. In conclusion, binding of the radioligand [125I] NKA to membranes from circular muscle is predominantly to the NK2 receptor. There may be a small component of binding to the NK1 receptor. The NK2 receptor mediates circular muscle contraction, whereas the role of the NK1 receptor in circular muscle is unclear. PMID- 10455256 TI - Mechanisms of gastroprotection by transdermal nitroglycerin in the rat. AB - Nitric oxide (NO) donors prevent experimentally-induced gastric mucosal damage, but their clinical utility is limited by short duration of action or unsuitability of the pharmaceutical form employed. This study analyses the gastroprotection elicited by a clinically used mode of continuous administration of an NO donor, namely the nitroglycerin patch. Application to rats of a transdermal patch that releases doses of nitroglycerin comparable to those used in man (40, 80, 160 and 400 ng min(-1) rat(-1)) reduced gastric damage induced by indomethacin (25 mg kg(-1), p.o. or s.c.). The nitroglycerin patch (160 ng min( 1) rat(-1)) also diminished damage by oral administration (1 ml) of acidified bile salts (100 mg kg(-1) taurocholic acid in 150 mM HCl) or 50% ethanol. Transdermal nitroglycerin (160 ng min(-1) rat(-1)) did not influence basal gastric blood flow, as measured by lasser-doppler flowmetry, but prevented its reduction by indomethacin. Transdermal nitroglycerin (160 ng min(-1) rat(-1)) prevented in vivo leukocyte rolling and adherence in the rat mesentery microvessels superfused with indomethacin, as evaluated by intravital microscopy. The transdermal nitroglycerin patch protects the gastric mucosa from damage by mechanisms that involve maintenance of mucosal blood flow and reduction of leukocyte-endothelial cell interaction. PMID- 10455257 TI - Inhibition of gelatinase activity in human airway epithelial cells and fibroblasts by dexamethasone and beclomethasone. AB - The effects of dexamethasone and beclomethasone on gelatinase activity released from lung epithelial cells (A549, NCI-H292 and Calu-3 cell lines and NHBE primary cultures) and human lung fibroblasts (HLF) were investigated. All cells spontaneously released gelatin-degrading activity but the amounts were unaffected by treatment with glucocorticoids. Phorbol myristate acetate (PMA) increased the amount of gelatinase activity in conditioned media prepared from all cell types examined. In epithelial cells, PMA induced the expression of gelatinase B, whereas in HLF the increased gelatinase activity resulted from increased activation of gelatinase A. Dexamethasone and beclomethasone produced concentration-dependent inhibition of PMA-induced gelatinase activity in HLF and epithelial cell lines. In the epithelial cell lines, the inhibition of activity was associated with an attenuation of enzyme induction by PMA. In contrast, primary cultures of human bronchial epithelial cells were unresponsive to dexamethasone at concentrations that were maximally effective at inhibiting gelatinase activity induced in other cells. PMID- 10455258 TI - Comparative antiplatelet effects of aspirin, vapiprost and GR144053, a GPIIb/IIIa antagonist, with a special reference to the role of platelet microaggregates. AB - Microthrombi produced have a potential to form larger thrombi, leading to vascular occlusions. Recently, a new device to easily detect microaggregates using laser-light scattering (LS) has been developed. We adopted this device to comparatively evaluate the inhibitory effects of aspirin (1,3 or 10 mg kg(-1)), vapiprost (0.3, 1 or 3 mg kg(-1)) or GR144053 (0.1, 0.3 or 1 mg kg(-1)) on ex vivo aggregation of hamster platelets in relation to their in vivo antithrombotic effects. A transluminal thrombus was produced in the hamster femoral artery by the photochemical reaction. Each compound was injected i.v. as a bolus 10 min prior to the reaction, showing a dose-dependent antithrombotic effect, i.e. they prolonged the time before the artery occluded. At that time cyclic flow reductions occurred more marked when aspirin or vapiprost was given. At the end of experiments, blood was collected to evaluate the platelet aggregation using both the new LS device and the conventional optical density (OD) method. Many more small aggregates were still formed when the highest dose of aspirin or vapiprost was used as compared with that of GR144053, although suppression of the platelet aggregation using the OD method, prolongation of the occlusion time and the bleeding time were quite similar. In conclusion, a GPIIb/IIIa antagonist markedly suppressed the microthrombi and reduced the cyclic flow reduction. This further indicates the importance of small aggregates as triggers of thrombosis and shows that prevention of their formation may result in improved vascular patency after thrombotic insult. PMID- 10455259 TI - Pharmacological characterization of extracellular acidification rate responses in human D2(long), D3 and D4.4 receptors expressed in Chinese hamster ovary cells. AB - This study characterized pharmacologically the functional responses to agonists at human dopamine D2(long) (hD2), D3 (hD3) and D4.4 (hD4) receptors separately expressed in cloned cells using the cytosensor microphysiometer. Dopaminergic receptor agonists caused increases in extracellular acidification rate in adherent Chinese hamster ovary (CHO) clones expressing hD2, hD3 or hD4 receptors. Acidification rate responses to agonists in other cell lines expressing these receptors were smaller than those in adherent CHO cells. The time courses and maximum increases in acidification rate of the agonist responses in adherent CHO cells were different between the three dopamine receptor clones. Responses were blocked by pretreatment of cells with pertussis toxin or amiloride analogues. Most agonists had full intrinsic activity at each of the dopamine receptor subtypes, as compared to quinpirole, however both enantiomers of UH-232 and (-)3 PPP were partial agonists in this assay system. The functional potency of full agonists at each of the three receptors expressed in CHO cells was either higher than, or similar to, the apparent inhibition constants (Ki) determined in [125I] iodosulpride competition binding studies. Functional selectivities of the agonists were less than radioligand binding selectivities. The rank orders of agonist potencies and selectivities were similar, but not identical, to the rank orders of radioligand binding affinities and selectivities. The dopamine receptor antagonists, iodosulpride and clozapine, had no effect on basal acidification rates but inhibited acidification responses in CHO cells to quinpirole in an apparently competitive manner. Antagonist potencies closely matched their radioligand binding affinities in these cells. PMID- 10455260 TI - Induction of eotaxin expression and release from human airway smooth muscle cells by IL-1beta and TNFalpha: effects of IL-10 and corticosteroids. AB - Eotaxin is a novel C-C chemokine with selective chemoattractant activity for eosinophils. We determined whether eotaxin could be produced by human airway smooth muscle (HASM) cells in culture and examined its regulation by interleukin 10 (IL-10) and the corticosteroid, dexamethasone. Stimulation of the cells with interleukin-1beta (IL-1beta) or tumour necrosis factor (TNFalpha) each at 10 ng ml(-1) induced the release of eotaxin protein with maximal accumulation by 24 h. Interferon-gamma (IFNgamma) alone at 10 ng ml(-1) had no effect and there was no synergy between these cytokines on the release of eotaxin. Reverse phase high performance liquid chromatographic (HPLC) analysis of supernatents from cells treated with TNFalpha (10 ng ml(-1) for 96 h showed immunoreactivity to eotaxin which eluted with the expected retention time of 34.5-35 min. Both IL-1beta and TNFalpha-induced release of eotaxin was not inhibited by dexamethasone (1 microM), however IL-10 (10 ng ml(-1)) had a significant inhibitory effect. Dexamethasone and IL-10 did not inhibit the induction of eotaxin mRNA induced by IL-1beta or TNFalpha. Thus, human airway smooth muscle cells can release eotaxin and could be an important source of chemokine production during airway inflammatory events. PMID- 10455261 TI - Involvement of cysteinyl leukotrienes in airway smooth muscle cell DNA synthesis after repeated allergen exposure in sensitized Brown Norway rats. AB - Airway smooth muscle thickening is a characteristic feature of airway wall remodelling in chronic asthma. We have investigated the role of the leukotrienes in airway smooth muscle (ASM) and epithelial cell DNA synthesis and ASM thickening following repeated allergen exposure in Brown Norway rats sensitized to ovalbumin. There was a 3 fold increase in ASM cell DNA synthesis, as measured by percentage bromodeoxyuridine (BrdU) incorporation, in repeatedly ovalbumin exposed (4.1%, 3.6-4.6; mean, 95% c.i.) compared to chronically saline-exposed rats (1.3%, 0.6-2.1; P<0.001). Treatment with a 5-lipoxygenase enzyme inhibitor (SB 210661, 10 mg kg(-1), p.o.) and a specific cysteinyl leukotriene (CysLT1) receptor antagonist, pranlukast (SB 205312, 30 mg kg(-1), p.o.), both attenuated ASM cell DNA synthesis. Treatment with a specific leukotriene B4 (BLT) receptor antagonist (SB 201146, 15 mg kg(-1), p.o.) had no effect. There was also a significant, 2 fold increase in the number of epithelial cells incorporating BrdU per unit length of basement membrane after repeated allergen exposure. This response was not inhibited by treatment with SB 210661, pranlukast or SB 201146. A significant increase in ASM thickness was identified following repeated allergen exposure and this response was attenuated significantly by SB 210661, pranlukast and SB 201146. Rats exposed to chronic allergen exhibited bronchial hyperresponsiveness to acetylcholine and had significant eosinophil recruitment into the lungs. Treatment with SB 210661, pranlukast or SB 201146 significantly attenuated eosinophil recruitment into the lungs, whilst having no significant effect on airway hyperresponsiveness. These data indicate that the cysteinyl leukotrienes are important mediators in allergen-induced ASM cell DNA synthesis in rats, while both LTB4 and cysteinyl leukotrienes contribute to ASM thickening and eosinophil recruitment following repeated allergen exposure. PMID- 10455262 TI - Effects of superoxide dismutase mimetics on the activity of nitric oxide in rat aorta. AB - A number of structurally distinct superoxide dismutase (SOD) mimetics were examined to determine if they shared the ability of authentic Cu/Zn SOD to produce endothelium-dependent relaxation of rings of rat aorta by protecting basal nitric oxide from destruction by endogenously produced superoxide anion. MnCl2 (10 nM-100 microM), CuSO4 (100 nM-1 mM) and CuDIPS (Cu [II] [diisopropylsalicylate]2; 100 nM-30 microM) each mimicked the ability of Cu/Zn SOD (0.1-300 u ml(-1)) to produce relaxation of phenylephrine-precontracted aortic rings in a manner inhibited by endothelial removal or treatment with N(G) nitro-L-arginine methyl ester (L-NAME, 100 microM). In contrast, MnTMPyP (Mn [III] tetrakis [1-methyl-4-pyridyl] porphyrin; 10 nM-30 microM) augmented phenylephrine-induced contraction and this was blocked by endothelial removal or treatment with L-NAME (100 microM), consistent with destruction rather than protection of basal nitric oxide activity. Pretreatment with Cu/Zn SOD (250 u ml( 1)) blocked this augmentation suggesting that it arose paradoxically through destruction of nitric oxide by superoxide anion. The spin trap agents tiron (100 nM-1 mM), tempol (100 nM-1 mM) and PTIYO (4-phenyl-2,2,5,5-tetramethyl imidazolin 1-yloxy-5-oxide; 100 nM-300 microM) all failed to promote endothelium-dependent relaxation. In fact, the last two augmented phenylephrine-induced tone and this was blocked by endothelial removal or treatment with L-NAME (100 microM), consistent with destruction of basal nitric oxide activity. This destruction was unaffected by pretreatment with Cu/Zn SOD (250 u ml(-1)) and probably reflected the direct ability of tempol and PTIYO to destroy nitric oxide. Thus, the ideal SOD mimetic for protection of nitric oxide activity in conditions of oxidant stress still awaits development. PMID- 10455263 TI - Expression of immediate early genes, GATA-4, and Nkx-2.5 in adrenergic-induced cardiac hypertrophy and during regression in adult mice. AB - Adrenoreceptor agonists induce a hypertrophic phenotype in vitro and in vivo. To investigate the molecular remodeling in chronic cardiac hypertrophy we infused adult male mice with vehicle. isoproterenol, phenylephrine or both agonists for 3, 7 or 14 days. All drugs increased cardiac mass. After minipump removal cardiac mass regressed to control levels within 7 days after PE and ISO treatment whereas ISO + PE treated hearts were incompletely regressed. ANF and beta-MHC, but not alpha-MHC, expression were increased by agonists at all time points. GATA-4, Nkx 2.5, Egr-1, c-jun and c-fos expression were increased after 3, 7 and 14 days of treatment. Expression was greatest after ISO+PE> >ISO>PE>vehicle infusion suggesting a synergistic effect of adrenoreceptor stimulation and indicating a greater effect of beta- than alpha-adrenergic action in vivo. After PE or ISO drug withdrawal the HW/BW was normal and Egr-1, c-jun, c-fos and GATA-4, but not Nkx2.5, expression dropped to control levels. HW/BW regression was incomplete after ISO+PE and elevated levels of Egr-1, c-jun and Nkx2.5 expression remained. A hydralazine-mediated reduction in blood pressure had no effect on the agonist induced cardiac hypertrophy or gene expression. In conclusion, we found that continued agonist stimulation, and not blood pressure. is responsible for the maintained increase in gene expression. Further, we found the decrease in gene expression in the regression after drug withdrawal was gene specific. PMID- 10455264 TI - The structural requirements for phorbol esters to enhance serotonin and acetylcholine release from rat brain cortex. AB - The effects of various phorbol-based protein kinase C (PKC) activators on the electrical stimulation-induced (S-I) release of serotonin and acetylcholine was studied in rat brain cortical slices pre-incubated with [3H]-serotonin or [3H] choline to investigate possible structure-activity relationships. 4beta-phorbol 12,13-dibutyrate (4betaPDB, 0.1-3.0 microM), enhanced S-I release of serotonin in a concentration-dependent manner whereas the structurally related inactive isomer 4alpha-phorbol 12, 13-dibutyrate (4alphaPDB) and phorbol 13-acetate (PA) were without effect. Another group of phorbol esters containing a common 13-ester substituent (phorbol 12,13-diacetate, PDA; phorbol 12-myristate 13-acetate, PMA; phorbol 12-methylaminobenzoate 13-acetate, PMBA) also enhanced S-I serotonin release with PMA being least potent. The deoxyphorbol monoesters, 12-deoxyphorbol 13-acetate (dPA), 12-deoxyphorbol 13-angelate (dPAng), 12-deoxyphorbol 13 phenylacetate (dPPhen) and 12-deoxyphorbol 13-isobutyrate (dPiB) enhanced S-I serotonin release but 12-deoxyphorbol 13-tetradecanoate (dPT) was without effect. The 20-acetate derivatives of dPPhen and dPAng were less effective in enhancing S I serotonin release compared to the parent compounds. With acetylcholine release all phorbol esters tested had a far lesser effect when compared to their facilitatory action on serotonin release with only 4betaPDB, PDA, dPA, dPAng and dPiB having significant effects. The effects of the phorbol esters on serotonin release were not correlated with their reported in vitro affinity and isozyme selectivity for PKC. A comparison across three transmitter systems (noradrenaline, dopamine, serotonin) suggests basic similarities in the structural requirements of phorbol esters to enhance transmitter release with short chain substituted mono- and diesters of phorbol being more potent facilitators of release than the long chain esters. Some compounds notably PDA, PMBA, dPPhen, dPPhenA had different potencies across noradrenaline, dopamine and serotonin. PMID- 10455265 TI - Studies on the role of serotonin receptor subtypes in the effect of sibutramine in various feeding paradigms in rats. AB - The effect of the 5-hydroxytryptamine (5-HT) and noradrenaline (NA) reuptake inhibitor sibutramine was studied in food deprived, neuropeptide Y (NPY)- or muscimol-injected rats. Sibutramine dose-dependently reduced feeding caused by food-deprivation (ED50 = 5.1+/-0.8 mg kg(-1)) or by NPY injection into the paraventricular nucleus of the hypothalamus (ED50 = 6.0+/-0.5 mg kg(-1)). The increase in food intake caused by muscimol injected into the dorsal raphe was not modified by sibutramine (1-10 mg kg(-1)). The hypophagic effect of 5.1 mg kg(-1) sibutramine in food-deprived rats was studied in rats pretreated with different serotonin receptor antagonists. Metergoline (non-selective, 0.3 and 1.0 mg kg( 1)), ritanserin (5-HT2A/2C, 0.5 and 1.0 mg kg(-1)) and GR127935 (5-HT1B/1D), 0.5 and 1.0 mg kg(-1)) did not modify the hypophagic effect of sibutramine, while SB206553 (5-HT2B/2C, 5 and 10 mg kg(-1)) slightly but significantly reduced it (Fint(2.53) = 3.4; P<0.05). The reduction in food intake caused by 6.0 mg kg(-1) sibutramine in NPY-injected rats was not modified by GR127935 (1.0 mg kg(-1)). The results suggest that, with the possible exception of a partial involvement of 5-HT2B/2C receptors in sibutramine's hypophagia in food-deprived rats, 5-HT1 and 5-HT2 receptor subtypes do not play an important role in the hypophagic effect of sibutramine, at least in the first 2 h after injection. PMID- 10455266 TI - Differential inhibitory mechanism of cyclic AMP on TNF-alpha and IL-12 synthesis by macrophages exposed to microbial stimuli. AB - Microbial stimuli such as bacterial lipopolysaccharide (LPS) or glycosylphosphatidylinositol-mucins derived from Trypanosoma cruzi trypomastigotes (tGPI-mucins) are effective stimulators of the synthesis of cytokines by macrophages. Here, we evaluated the ability of cyclic AMP mimetic or elevating agents to modulate TNF-alpha and IL-12 synthesis by murine inflammatory macrophages. Cholera Toxin (ChTx) inhibited tGPI-mucins (2.5 nM) or LPS (100 ng ml(-1)) induced TNF-alpha and IL-12(p40) synthesis in a concentration-dependent manner. Similarly, the cyclic AMP mimetics, 8-bromo cyclic AMP or dibutyryl cyclic AMP, or prostaglandin (PG) E2 inhibited the synthesis of both cytokines by macrophages exposed to microbial stimuli. The protein kinase A inhibitor H-89 partially reversed the inhibitory effects of dibutyryl cyclic AMP and PGE2 on both IL-12(p40) and TNF-alpha synthesis. Pretreatment of macrophages with dibutyryl cyclic AMP or ChTx augmented the synthesis of IL-10 triggered by microbial products. Elevation of cyclic AMP inhibited the synthesis of TNF-alpha, but not IL-12(p40), by inflammatory macrophages from IL-10 knockout mice. Kinetic studies showed that synthesis of both TNF-alpha and IL-10 peaked at 8 h and IL-12 at 24 h after stimulation with microbial stimuli. Together, our findings favour the hypothesis that the cyclic AMP inhibitory activity on IL-12(p40) but not on TNF-alpha synthesis is dependent on de novo protein synthesis, most likely involving IL-10, by macrophages stimulated with microbial products. Accordingly, dibutyryl cyclic AMP inhibited IL-12(p40) synthesis only when added before or at the same time of the stimuli. In contrast, the effect of this cyclic AMP analogue on TNF-alpha synthesis was protracted and observed even 2 h after the addition of the stimuli. PMID- 10455267 TI - The antiplatelet activity of PMC, a potent alpha-tocopherol analogue, is mediated through inhibition of cyclo-oxygenase. AB - PMC, a potent alpha-tocopherol derivative, dose-dependently (5-25 microM) inhibited the ATP-release reaction and platelet aggregation in washed human platelets stimulated by agonists (collagen and ADP). PMC also dose-dependently inhibited the intracellular Ca2+ mobilization, whereas it did not inhibit phosphoinositide breakdown in human platelets stimulated by collagen. PMC (10 and 25 microM) significantly inhibited collagen-stimulated thromboxane A2 (TxA2) formation in human platelets. On the other hand, PMC (25 and 100 microM) did not increase the formation of cyclic AMP or cyclic GMP in platelets. Moreover, PMC (25, 100, and 200 microM) did not affect the thromboxane synthetase activity of aspirin-treated platelet microsomes. PMC (10 and 25 microM) markedly inhibited the exogenous arachidonic acid (100 microM)-induced prostaglandin E2 (PGE2) formation in the presence of imidazole (600 microM) in washed human platelets, indicating that PMC inhibits cyclo-oxygenase activity. We conclude that PMC may exert its anti-platelet aggregation activity by inhibiting cyclooxygenase activity, which leads to reduced prostaglandin formation; this, in turn, is followed by a reduction of TxA2 formation, and finally inhibition of [Ca2+]i mobilization and ATP-release. PMID- 10455269 TI - Role of superoxide dismutase in in vivo and in vitro nitrate tolerance. AB - We assessed whether pharmacological inhibition of CuZn-superoxide dismutase (SOD) mimics the molecular mechanism of either in vitro or in vivo nitrovasodilator tolerance. In endothelium-intact aortic rings from in vivo tolerant rabbits the GTN- and acetylcholine (ACh)-induced maximal relaxation was attenuated by 36 and 23%, respectively. In vitro treatment of control rings with GTN (1 h 10 microM) similarly attenuated the vasorelaxant response to GTN, but not to ACh. Formation of superoxide radicals (*O2-) in endothelium-intact rings (lucigenin chemiluminescence) increased 2.5 fold in in vivo tolerance, but significantly decreased in in vitro tolerance. The membrane associated NADH oxidase activity was increased 2.5 fold in homogenates of in vivo tolerant aortae, but was not changed in in vitro tolerant aorta. Conversely, SOD activity and protein expression was halved in in vivo tolerance, but SOD activity was not altered by in vitro tolerance. The *O2- scavenger tiron (10 mM) effectively restored the vasorelaxant response to GTN in in vivo tolerant aortic rings, but not the reduced response to GTN in in vitro tolerant rings. Pretreatment (1 h) of vessels with diethyldithiocarbamate (DETC; 10 mM) attenuated vasorelaxant responses to GTN and ACh, increased vascular *O2- production, and inhibited SOD activity in vessel homogenates to a similar degree as observed in in vivo tolerance. DETC treatment of in vivo-tolerant vessels induced an additional increase in *O2- production. Increased *O2- production in in vivo nitrate tolerant aorta is associated with activation of vascular NADH oxidase and inactivation of CuZnSOD. Therefore, in vivo tolerance can be mimicked by in vitro inhibition of CuZnSOD, but not by in vitro exposure to GTN, which does not affect vascular *O2- production, NADH oxidase and CuZnSOD. PMID- 10455268 TI - Separation of M-like current and ERG current in NG108-15 cells. AB - Differentiated NG108-15 neuroblastoma x glioma hybrid cells were whole-cell voltage-clamped. Hyperpolarizing pulses, superimposed on a depolarized holding potential (-30 or -20 mV), elicited deactivation currents which consisted of two components, distinguishable by fitting with two exponential functions. Linopirdine [DuP 996, 3,3-bis(4-pyridinylmethyl)-1-phenylindolin-2-one), a neurotransmitter-release enhancer known as potent and selective blocker of the M current of rat sympathetic neurons, in concentrations of 5 or 10 microM selectively inhibited the fast component (IC50 = 14.7 microM). The slow component was less sensitive to linopirdine (IC50>20 microM). The class III antiarrhythmics [(4-methylsulphonyl)amido]benzenesulphonamide (WAY-123.398) and 1-[2-(6-methyl-2 pyrydinil)ethyl]-4-(4-methylsulphonylaminobenz oyl) piperidine (E-4031), selective inhibitors of the inwardly rectifying ERG (ether-a-go-go-related gene) potassium channel, inhibited predominantly the slow component (IC50 = 38 nM for E 4031). The time constant of the WAY-123.398-sensitive current resembled the time constant of the slow component in size and voltage dependence. Inwardly rectifying ERG currents, recorded in K+ -rich bath at strongly negative pulse potentials, resembled the slow component of the deactivation current in their low sensitivity to linopirdine (28% inhibition at 50 microM). The size of the slow component varied greatly between cells. Accordingly, varied the effect of WAY 123.398 on deactivation current and holding current. RNA transcripts for the following members of the ether-a-go-go gene (EAG) K+ channel family were found in differentiated NG108-15 cells: ERG1, ERG2, EAGI, EAG-like (ELK)1, ELK2; ERG3 was only present in non-differentiated cells. In addition, RNA transcripts for KCNQ2 and KCNQ3 were found in differentiated and non-differentiated cells. We conclude that the fast component of the deactivation current is M-like current and the slow component is deactivating ERG current. The molecular correlates are probably KCNQ2/KCNQ3 and ERG1/ERG2, respectively. PMID- 10455270 TI - Dual mode of stimulation by the beta-carboline ZK 91085 of recombinant GABA(A) receptor currents: molecular determinants affecting its action. AB - In electrophysiological measurements the beta-carboline ethyl 6-benzyloxy-beta carboline-3-carboxylate (ZK 91085) acts as a positive allosteric modulator on rat recombinant alpha1beta2gamma2 GABA(A) receptors and binds with high affinity (IC50-1.5 nM) to the [3H]-flunitrazepam site. Flumazenil was able to partially counteract the current modulation. These observations indicate an action of ZK 91085 at the benzodiazepine binding site. At the dual subunit combination alpha1beta2, which lacks the gamma subunit required for benzodiazepine modulation, we still observed a potentiation of GABA currents. Thus ZK 91085 acts via an additional site on the channel. At the subunit combination alpha1beta1, ZK 91085 potentiation is strongly reduced as compared to alpha1beta2. In binding studies, ZK 91085 was able to decrease [35S]-TBPS binding in alpha1beta2gamma2 and alpha1beta2 but not in alpha1beta1. This selectivity of ZK 91085 for receptors containing the beta2 isoform over those containing the beta1 isoform is reminiscent of the action of loreclezole. To identify amino acid residues important for the second type of modulation, we functionally compared wild type alpha1beta2 and mutant receptors for stimulation by ZK 91085. The mutation beta2N265S, that abolishes loreclezole effects, also abolishes ZK 91085 stimulation. The mutation beta2Y62L increased stimulation by ZK 91085 3-4 fold, locating an influencing entity of the second type of action of ZK 91085 at an alpha/beta subunit interface. Structural intermediates of ZK 91085 and the beta carboline abecarnil, the latter of which only slightly potentiated GABA currents in alpha1/beta2, were analysed to determine structural requirements for modulation. ZK 91085 thus allosterically stimulates the GABA(A) receptor through two sites of action: the benzodiazepine site and the loreclezole site in contrast to classical beta-carbolines, that confer negative allosteric modulation through the benzodiazepine site. PMID- 10455271 TI - Subtype-selective inhibition of [methyl-3H]-N-methylscopolamine binding to muscarinic receptors by alpha-truxillic acid esters. AB - Seven esters of alpha-truxillic acid have been synthesized: bis-3-piperidylpropyl ester and its quaternary bis-N-ethyl derivative, bis-N-diethylaminopropyl ester and its quaternary bis-N-methyl derivative, and bis-4-piperidylbutyl ester and its quaternary bis-N-methyl and bis-N-ethyl derivatives. All esters inhibited the specific binding of muscarinic receptor antagonist [methyl-3H]-N methylscopolamine ([3H]-NMS) to muscarinic receptors in membranes of CHO cell lines stably expressing the human gene for the M1, M2, M3 or M4 subtype of muscarinic receptors. All esters displayed the highest potency at the M2 and the lowest potency at the M3 receptor subtype. In experiments performed on the M2 muscarinic receptor subtype, the affinity between the receptors and the esters was greatly increased when the concentration of ions was diminished. The highest affinities were found for the tertiary bis-3-piperidylpropyl and bis-4 piperidylbutyl aminoesters (equilibrium dissociation constants of 52 and 179 pM, respectively, in the low ionic strength medium). All investigated esters slowed down the dissociation of [3H]-NMS from the M2 muscarinic receptor subtype. [3H] NMS dissociation from the M1, M3 and M4 muscarinic receptor subtypes was investigated in experiments with the bis-4-piperidylbutyl aminoester and also found to be decelerated. It is concluded that the esters of alpha-truxillic acid act as M2-selective allosteric modulators of muscarinic receptors and that, by their potency, the tertiary bis-3-piperidylpropyl and bis-4-piperidylbutyl aminoesters surpass the other known allosteric modulators of these receptors. PMID- 10455272 TI - Functional characterization and m-RNA expression of 5-HT receptors mediating contraction in human umbilical artery. AB - 5-HT1-like and 5-HT2 receptors have both been described to mediate contractions to 5-HT in the human umbilical artery (HUA). However, the nature of the 5-HT receptor subtypes is unknown. 2 In isometric force studies with ring preparations of HUA alpha-methyl-5-hydroxytryptamine (alpha-Me-5-HT) and 5-hydroxytryptamine (5-HT) contracted HUA with pED50 values of 8.04 and 7.74, respectively. In the presence of a subthreshold concentration of another vasoconstrictor sumatriptan and 5-nonyloxytryptamine elicited concentration-dependent contractions with pEC50 values of 7.21 and 7.67, respectively. In the presence of the selective 5-HT1B/D receptor antagonist GR127935, contractile responses elicited by sumatriptan and 5 nonyloxytryptamine were competitively antagonized (pKB 9.01 and 9.02, respectively). In the experiments with 5-HT, GR127935 appeared to be non competitive with shallow Schild plot slopes. The data were fitted with two linear regression lines and the calculated pKB of the high affinity component (8.90) was comparable to that expected for GR127935 at the 5-HT1B/1D receptor. Several 5-HT2 selective receptor antagonists (spiperone, cyproheptadine, pirenperone) competitively inhibited responses to 5-HT. The selective 5-HT2A antagonist ketanserin against sumatriptan and 5-nonyloxytryptamine behaved as a weak antagonist while against 5-HT demonstrated a competitive antagonism (pKB 8.56). Using specific primers for human 5-HT1B, 5-HT1D and 5-HT2A receptor genes, the reverse transcriptase-polymerase chain reaction revealed mRNA expression of 5 HT1B and 5-HT2A receptors in the HUA. The results suggest that the HUA has a functional population of 5-HT1B and 5-HT2A receptor subtypes which are involved in the contractile response to 5-HT. Contractions mediated by 5-HT1B receptors can be 'uncovered' by exposure to other vasoactive agents. PMID- 10455273 TI - Prejunctional angiotensin receptors involved in the facilitation of noradrenaline release in mouse tissues. AB - The effect of angiotensin II, angiotensin III, angiotensin IV and angiotensin-(1 7) on the electrically induced release of noradrenaline was studied in preparations of mouse atria, spleen, hippocampus, occipito-parietal cortex and hypothalamus preincubated with [3H]-noradrenaline. The prejunctional angiotensin receptor type was investigated using the non-selective receptor antagonist saralasin (AT1/AT2) and the AT1 and AT2 selective receptor antagonists losartan and PD 123319, respectively. In atrial and splenic preparations, angiotensin II (0.01 nM-0.1 microM) and angiotensin III (0.01 and 0.1 nM-1 microM) increased the stimulation-induced overflow of tritium in a concentration-dependent manner. Angiotensin IV, only at high concentrations (1 and 10 pM), enhanced tritium overflow in the atria, while angiotensin-(1-7) (0.1 nM-10 microM) was without effect in both preparations. In preparations of hippocampus, occipito-parietal cortex and hypothalamus, none of the angiotensin peptides altered the evoked overflow of tritium. In atrial and splenic preparations, saralasin (0.1 microM) and losartan (0.1 and 1 microM), but not PD 123319 (0.1 microM), shifted the concentration-response curves of angiotensin II and angiotensin III to the right. In conclusion, in mouse atria and spleen, angiotensin II and angiotensin III facilitate the action potential induced release of noradrenaline via a prejunctional AT1 receptor. Only high concentrations of angiotensin IV are effective in the atria and angiotensin-(1-7) is without effect in both preparations. In mouse brain areas, angiotensin II, angiotensin III, angiotensin IV and angiotensin-(1-7) do not modulate the release of noradrenaline. PMID- 10455274 TI - Pharmacological evidence that alpha1-and alpha2-adrenoceptors mediate vasoconstriction of carotid arteriovenous anastomoses in anaesthetized pigs. AB - Vasoconstriction of carotid arteriovenous anastomoses may be involved in the therapeutic action of acutely acting anti-migraine agents, including the triptans and ergot alkaloids. While 5-HT1B/1D receptors mediate the effect of triptans, ergotamine and dihydroergotamine also interact with alpha-adrenoceptors. In the present study, we investigated the potential role of alpha1- and alpha2 adrenoceptors in mediating vasoconstriction of carotid arteriovenous anastomoses in anaesthetized pigs. Ten minute intracarotid infusions of phenylephrine (1, 3 and 10 microg kg(-1) min(-1)) or BHT 933 (3, 10 and 30 microg kg(-1) min(-1)) produced dose-dependent decreases in total carotid and arteriovenous anastomotic conductances; no changes were observed in the capillary fraction. The carotid vascular effects of phenylephrine and BHT 933 were selectively abolished by prazosin (100 microg kg(-1), i.v.) and rauwolscine (300 microg kg(-1), i.v.), respectively. The responses to phenylephrine and BHT 933 were not affected by the selective 5-HT1B/1D receptor antagonist GR127935 (500 microg kg(-1), i.v.). These results show that both alpha1- and alpha2-adrenoceptors can mediate vasoconstriction of carotid arteriovenous anastomoses in anaesthetized pigs. Since vasoconstrictor activity in this in vivo model is predictive of anti migraine activity, an agonist activity at particularly the alpha2-adrenoceptor subtypes, in view of their less ubiquitous nature, could provide migraine abortive potential. Thus, the present results may aid further understanding of the mode of action of some current anti-migraine agents and may eventually be helpful in the development of future treatment in migraine. PMID- 10455275 TI - Quantitation of extracellular UTP using a sensitive enzymatic assay. AB - The wide distribution of the uridine nucleotide-activated P2Y2, P2Y4 and P2Y6 receptors suggests a role for UTP as an important extracellular signalling molecule. However, direct evidence for UTP release and extracellular accumulation has been addressed only recently due to the lack of a sensitive assay for UTP mass. In the present study, we describe a method that is based on the uridinylation of [14C]-glucose-1P by the enzyme UDP-glucose pyrophosphorylase which allows quantification of UTP in the sub-nanomolar concentration range. The UTP-dependent conversion of [14C]-glucose-1P to [14C]-UDP-glucose was made irreversible by including the pyrophosphate scavenger inorganic pyrophosphatase in the reaction medium and [14C]-glucose-1P and [14C]-UDP-glucose were separated and quantified by HPLC. Formation of [14C]-UDP-glucose was linearly observed between 1 and 300 nM UTP. The reaction was highly specific for UTP and was unaffected by a 1000 fold molar excess of ATP over UTP. Release of UTP was measured with a variety of cells including platelets and leukocytes, primary airway epithelial cells, rat astrocytes and several cell lines. In most resting attached cultures, extracellular UTP concentrations were found in the low nanomolar range (1-10 nM in 0.5 ml medium bathing 2.5 cm2 dish). Up to a 20 fold increase in extracellular UTP levels was observed in cells subjected to a medium change. Extracellular UTP levels were 10-30% of the ATP levels in both resting and mechanically-stimulated cultured cells. In unstirred platelets, a 1:100 ratio UTP/ ATP was observed. Extracellular UTP and ATP increased 10 fold in thrombin stimulated platelets. Detection of UTP in nanomolar concentrations in the medium bathing resting cultures suggests that constitutive release of UTP may provide a mechanism of regulation of the basal activity of uridine nucleotide sensitive receptors. PMID- 10455276 TI - Multiple effector pathways regulate the insulin secretory response to the imidazoline RX871024 in isolated rat pancreatic islets. AB - When isolated rat islets were cultured for 18 h prior to use, the putative imidazoline binding site ligand, RX871024 caused a dose-dependent increase in insulin secretion at both 6 mM and 20 mM glucose. By contrast, a second ligand, efaroxan, was ineffective at 20 mM glucose whereas it did stimulate insulin secretion in response to 6 mM glucose. Exposure of islets to RX871024 (50 microM) for 18 h, resulted in loss of responsiveness to this reagent upon subsequent re exposure. However, islets that were unresponsive to RX871024 still responded normally to efaroxan. The imidazoline antagonist, KU14R, blocked the insulin secretory response to efaroxan, but failed to prevent the stimulatory response to RX871024. By contrast with its effects in cultured islets, RX871024 inhibited glucose-induced insulin release from freshly isolated islets. Efaroxan did not inhibit insulin secretion under any conditions studied. In freshly isolated islets, the effects of RX871024 on insulin secretion could be converted from inhibitory to stimulatory, by starvation of the animals. Inhibition of insulin secretion by RX871024 in freshly isolated islets was prevented by the cyclo oxygenase inhibitors indomethacin or flurbiprofen. Consistent with this, RX871024 caused a marked increase in islet PGE2 formation. Efaroxan did not alter islet PGE2 levels. The results suggest that RX871024 exerts multiple effects in the pancreatic beta-cell and that its effects on insulin secretion cannot be ascribed only to interaction with a putative imidazoline binding site. PMID- 10455277 TI - Ligand efficacy and potency at recombinant human MT2 melatonin receptors: evidence for agonist activity of some mt1-antagonists. AB - NIH3T3 fibroblast cells transfected with the full-length coding region of the MT2 human melatonin receptor stably expressed the receptor that is coupled to a pertussis toxin-sensitive G protein and exhibits high affinity for melatonin (K(I) = 261 pM). The order of apparent affinity for selected compounds was: 4 phenyl-2-propionamidotetralin (4P-PDOT) > 2-phenylmelatonin > 2-iodomelatonin > 2 bromomelatonin > 6-chloromelatonin > or = melatonin > luzindole > N-acetyl tryptamine > or = N-[(2-phenyl-1H-indol-3-yl)ethyl]cyclobutanecarboxamide (compound 6) > N-acetylserotonin. 4P-PDOT exhibited a very high selectivity (approximately 22,000 times) for the MT2 receptor with respect to the mt1 receptor subtype, as tested in comparative experiments with membrane preparations from NIH3T3 cells stably transfected with the human mt1 receptor. MT2 melatonin receptors mediated incorporation of [35S]-GTPgammaS into isolated membranes via receptor catalyzed exchange of [35S]-GTPgammaS for GDP. The relative intrinsic activity and potency of the compounds were subsequently studied by using [35S] GTPgammaS incorporation. The order of potency was equal to the order of apparent affinity. Melatonin and full agonists increased [35S]-GTPgammaS binding by 250% over basal (taken as 100%). Luzindole did not increase basal [35S]-GTPgammaS binding but competitively inhibited melatonin-stimulated [35S]-GTPgammaS binding, thus exhibiting antagonist action. The other two mt1 antagonists used here, 4P PDOT and N-[(2-phenyl-1H-indol-3-yl)ethyl]cyclobutanecarboxamide, behaved as partial agonists at the MT2 subtype, with relative intrinsic activities of 0.37 and 0.39, respectively. These findings show, for the first time, important differences in the intrinsic activity of analogues between the human mt1 and MT2 melatonin receptor subtypes. PMID- 10455279 TI - Cross-resistance with dieldrin of a novel tricyclic dinitrile GABA receptor antagonist. AB - A novel tricyclic dinitrile, KN244, blocked the wild-type (dieldrin-sensitive) homo-oligomeric gamma-aminobutyric acid (GABA)-gated chloride channel of Drosophila melanogaster expressed in Xenopus oocytes. Sensitivity to the block by KN244 of the response to 30 microM GABA (IC50=41.6 nM, wild-type RDLac) was reduced abut 100 fold (IC50=4.5 microM) in the dieldrin-resistant (RDLacA302S) form of RDL. PMID- 10455278 TI - IL-12 as a therapeutic target for pharmacological modulation in immune-mediated and inflammatory diseases: regulation of T helper 1/T helper 2 responses. AB - Interleukin-12 (IL-12) is a pivotal cytokine in driving the immune system towards a T helper (Th)1 type response and preventing a Th2 type immune profile. Therefore, IL-12 is indispensable in the defense against certain, mainly intracellular pathogens, but overproduction of this cytokine is crucially involved in the etiology of several inflammatory and autoimmune diseases. Hence, IL-12 is an ideal target for pharmacological intervention in the therapy of autoimmune and inflammatory diseases. The production of IL-12 and a resultant Th1 type immune response can be suppressed with several pharmacological approaches including modulation of intracellular cyclic AMP levels, glucocorticoids and nuclear factor-kappaB inhibition. IL-12 responsiveness may be inhibited using anti-IL-12 antibodies, soluble IL-12 receptors or the IL-12 p40 homodimer. Exploitation of these approaches may provide novel means for the experimental therapy of a variety of pathophysiological states. PMID- 10455280 TI - Participation of protein kinases in staurosporine-induced interleukin-6 production by rat peritoneal macrophages. AB - The incubation of rat peritoneal macrophages in the presence of staurosporine, a non-specific protein kinase inhibitor, induced interleukin-6 (IL-6) production in a time- and concentration-dependent manner at 6.3-63 nM, but at 210 nM, the stimulant effect on IL-6 production was reduced. The levels of IL-6 mRNA as determined by a reverse transcription-polymerase chain reaction were also increased by staurosporine in parallel with the ability to induce IL-6 production. Compounds structurally related to staurosporine including K-252a (non specific protein kinase inhibitor) and KT-5720 (inhibitor of cyclic AMP-dependent protein kinase, PKA), did not increase IL-6 production by peritoneal macrophages. Staurosporine-induced increases in IL-6 production and expression of IL-6 mRNA were decreased by the PKC inhibitors, H-7 (2.7-27 microM), Ro 31-8425 (1-10 microM) and calphostin C (0.3-3 microM) and by the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor LY294002 (30-100 microM), but were further increased by the protein tyrosine kinase (PTK) inhibitor, genistein (12-37 microM). The staurosporine-induced increase in IL-6 production was not affected by the PKA inhibitor, H-89 (0.1-3 microM). These findings suggest that the induction of IL-6 production by staurosporine is secondary to elevation of IL-6 mRNA level, which, in turn, is positively regulated by the activation of PKC and PI 3-kinase and negatively regulated by the activation of PTK. PKA does not appear to play a significant role. PMID- 10455281 TI - Protection by imidazol(ine) drugs and agmatine of glutamate-induced neurotoxicity in cultured cerebellar granule cells through blockade of NMDA receptor. AB - This study was designed to assess the potential neuroprotective effect of several imidazol(ine) drugs and agmatine on glutamate-induced necrosis and on apoptosis induced by low extracellular K+ in cultured cerebellar granule cells. Exposure (30 min) of energy deprived cells to L-glutamate (1-100 microM) caused a concentration-dependent neurotoxicity, as determined 24 h later by a decrease in the ability of the cells to metabolize 3-(4,5-dimethythiazol-2-yl)-2,5 diphenyltetrazoliumbromide (MTT) into a reduced formazan product. L-glutamate induced neurotoxicity (EC50=5 microM) was blocked by the specific NMDA receptor antagonist MK-801 (dizocilpine). Imidazol(ine) drugs and agmatine fully prevented neurotoxicity induced by 20 microM (EC100) L-glutamate with the rank order (EC50 in microM): antazoline (13)>cirazoline (44)>LSL 61122 [2-styryl-2-imidazoline] (54)>LSL 60101 [2-(2-benzofuranyl) imidazole] (75)>idazoxan (90)>LSL 60129 [2 (1,4-benzodioxan-6-yl)-4,5-dihydroimidazole](101)>RX82 1002 (2-methoxy idazoxan) (106)>agmatine (196). No neuroprotective effect of these drugs was observed in a model of apoptotic neuronal cell death (reduction of extracellular K+) which does not involve stimulation of NMDA receptors. Imidazol(ine) drugs and agmatine fully inhibited [3H]-(+)-MK-801 binding to the phencyclidine site of NMDA receptors in rat brain. The profile of drug potency protecting against L-glutamate neurotoxicity correlated well (r=0.90) with the potency of the same compounds competing against [3H]-(+)-MK-801 binding. In HEK-293 cells transfected to express the NR1-1a and NR2C subunits of the NMDA receptor, antazoline and agmatine produced a voltage- and concentration-dependent block of glutamate induced currents. Analysis of the voltage dependence of the block was consistent with the presence of a binding site for antazoline located within the NMDA channel pore with an IC50 of 10-12 microM at 0 mV. It is concluded that imidazol(ine) drugs and agmatine are neuroprotective against glutamate-induced necrotic neuronal cell death in vitro and that this effect is mediated through NMDA receptor blockade by interacting with a site located within the NMDA channel pore. PMID- 10455283 TI - Antagonist activities of mecamylamine and nicotine show reciprocal dependence on beta subunit sequence in the second transmembrane domain. AB - We show that a portion of the TM2 domain regulates the sensitivity of beta subunit-containing rat neuronal nicotinic AChR to the ganglionic blocker mecamylamine, such that the substitution of 4 amino acids of the muscle beta subunit sequence into the neuronal beta4 sequence decreases the potency of mecamylamine by a factor of 200 and eliminates any long-term effects of this drug on receptor function. The same exchange of sequence that decreases inhibition by mecamylamine produces a comparable potentiation of long-term inhibition by nicotine. Inhibition by mecamylamine is voltage-dependent, suggesting a direct interaction of mecamylamine with sequence elements within the membrane field. We have previously shown that sensitivity to TMP (tetramethylpiperidine) inhibitors is controlled by the same sequence elements that determine mecamylamine sensitivity. However, inhibition by bis-TMP compounds is independent of voltage. Our experiments did not show any influence of voltage on the inhibition of chimeric receptors by nicotine, suggesting that the inhibitory effects of nicotine are mediated by binding to a site outside the membrane's electric field. An analysis of point mutations indicates that the residues at the 6' position within the beta subunit TM2 domain may be important for determining the effects of both mecamylamine and nicotine in a reciprocal manner. Single mutations at the 10' position are not sufficient to produce effects, but 6' 10' double mutants show more effect than do the 6' single mutants. PMID- 10455282 TI - Involvement of 5-HT1B receptors in collar-induced hypersensitivity to 5 hydroxytryptamine of the rabbit carotid artery. AB - In humans intimal thickening is aprerequisite of atherosclerosis. Application of a silicone collar around the rabbit carotid artery induces an intimal thickening but in addition it increases the sensitivity to the vasoconstrictor action of serotonin (5-hydroxytryptamine, 5-HT). The 5-HT receptors involved in collar induced hypersensitivity to 5-HT were investigated using several agonists and antagonists. One week after placement of collars around both carotid arteries of anaesthetized rabbits, rings (2 mm width) from inside (=collar) and outside (=sham) the collars were mounted in organ baths (10 ml) for isometric force measurements at 6 g loading tension. Collared rings were more sensitive to the contractile effect of 5-HT (7.6 fold) and 5-carboxamidotryptamine (31 fold, 5-CT, 5-HT1 agonist) in cumulative concentration response curves. Sumatriptan (5 HT1B/1D agonist) caused concentration-dependent constrictions in collared rings only. Collar placement did not significantly alter pA2 values (Schild regression) or apparent pKb values (non-linear regression) of spiperone and methysergide (mixed 5-HT2A/5-HT1 antagonists) or ketanserin and ritanserin (5-HT2A antagonists), indicating unchanged binding characteristics of the 5-HT2A receptor. However, the reduced slope of the Schild regression pointed to a heterogeneous receptor population in collared rings. In contrast, the apparent pKb value of methiothepin (5-HT1B antagonist) was significantly reduced by collar placement, and its antagonism shifted from non-surmountable in sham rings to surmountable in collared segments. Taken together, this study demonstrates that the serotonergic receptor involved in the hypersensitivity to 5-HT of rabbit collared carotid artery is a 5-HT1B receptor subtype. PMID- 10455284 TI - Mutation at the putative GABA(A) ion-channel gate reveals changes in allosteric modulation. AB - We have mutated a conserved leucine in the putative membrane-spanning domain to serine in human GABA(A) beta2 and investigated the actions of a number of GABA(A) agonists, antagonists and modulators on human alpha1beta2deltaL259Sgamma2s compared to wild type alpha1beta2gamma2s GABA(A) receptors, expressed in Xenopus oocytes. The mutation resulted in smaller maximum currents to gamma-aminobutyric acid (GABA) compared to alpha1beta2gamma2s receptors, and large leak currents resulting from spontaneous channel opening. As reported, this mutation significantly decreased the GABA EC50 (110 fold), and reduced desensitization. Muscimol and the partial agonists 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP) and piperidine-4-sulphonic acid (P4S) also displayed a decrease in EC50. In addition to competitively shifting GABA concentration response curves, the antagonists bicuculline and SR95531 both inhibited the spontaneous channel activity on alpha1beta2deltaL259Sgamma2s receptors, with different degrees of maximum inhibition. The effects of a range of allosteric modulators, including benzodiazepines and anaesthetics were examined on a submaximal GABA concentration (EC20). Compared to wild type, none of these modulators potentiated the EC20 response of alpha1beta2deltaL259Sgamma2s receptors, however they all directly activated the receptor in the absence of GABA. To conclude, the above mutation resulted in receptors which exhibit a degree of spontaneous activity, and are more sensitive to agonists. Benzodiazepines and other agents modulate constitutive activity, but positive modulation of GABA is lost. The competitive antagonists bicuculline and SR95531 can also act as allosteric channel modulators through the same GABA binding site. PMID- 10455285 TI - Differential effects of adenine nucleotide analogues on shape change and aggregation induced by adnosine 5-diphosphate (ADP) in human platelets. AB - Adenosine 5'-diphosphate (ADP) induces human blood platelets to aggregate and change shape, and it has been suggested that these two responses are mediated by more than one subtype of ADP receptor. The structure-activity relationships for several analogues of adenine nucleotides in causing aggregation and shape change were measured and compared in washed platelets using an aggregometer. ADP and its analogues 2-methylthioadenosine 5'-diphosphate (2-methylthio-ADP), adenosine 5'(alpha,beta-methylene)diphosphonate (AMPCP), S(P)-adenosine 5'-O-(1 thiodiphosphate) (AD-P alphaS) and adenosine 5'-O-(2-thiodiphosphate) (ADPbetaS) were used as agonists. Adenosine 5'-triphosphate (ATP) and its analogues, P1, P5 diadenosine pentaphosphate (ApsA), adenosine (5'-(alpha,beta methylene)triphosphonate (AMPCPP), 2-methylthioadenosine 5'-triphosphate (2 methylthio-ATP) and uridine 5'-triphosphate (UTP), as well as the trypanocidal drug suramin, were used as antagonists. In general, the structure-activity relationships for both responses were similar, but for some analogues differences were observed. ADPalphaS and ADPbetaS were much more potent agonists relative to ADP for shape change than for aggregation and indeed ADPalphaS antagonized ADP induced aggregation with an apparent pK(B) value of 5.5+/-0.1. 2-Methylthio-ATP also had different effects in aggregation and shape change, being a much higher affinity antagonist of aggregation than of shape change with an apparent pK(B) value of 7.0+/-0.2 for aggregation and 5.2+/-0.2 for shape change. These results support the suggestion that these two responses are mediated by multiple ADP receptors on human platelets, and are consistent with shape change being mediated via one receptor (the P2Y1 receptor) with aggregation requiring the activation of two receptors (the P2Y1 and another P2Y receptor). PMID- 10455286 TI - Evaluation of alpha1-adrenoceptors in the rabbit iris: pharmacological characterization and expression of mRNA. AB - Subtypes of alpha1-adrenoceptor in rabbit iris have been examined in functional, binding and molecular biological experiments. In functional studies, exogenous and endogenous noradrenaline produced contractions of the iris dilator muscle. The contractile responses to noradrenaline were competitively antagonized by a range of alpha1-adrenoceptor antagonists (pA2 values): prazosin (8.1), WB4101 (8.2), BMY7378 (5.9), YM617 (9.5), JTH-601 (8.8), HV723 (7.8) and KMD-3213 (9.8). The same order of inhibitory potency was seen in the adrenergic responses to electrical stimulation. This affinity profile corresponds well to that of the putative alpha1L-adrenoceptor, which has been proposed in lower urinary tract tissues. In binding studies on rabbit iris membrane however, prazosin, KMD-3213 and WB4101 displayed high affinity (pKd or pKi: 9.6, 10.3, 9.6, respectively), and BMY7378 displayed low affinity (pKi: 6.9). These results show that the binding sites typically correspond to alpha1A-adrenoceptor subtype in character, and we could not detect the significant amount of alpha1L-adrenoceptor subtype. The expression of the three distinct mRNAs that encode proteins of alpha1a-, alpha1b- and alpha1d-adrenoceptors was studied using reverse transcription polymerase chain reaction (RT-PCR). RT-PCR demonstrated the strongest expression of the alpha1a-adrenoceptor, weak expression of the alpha1b-adrenoceptor and undetectable expression of the alpha1d-adrenoceptor. The present study suggests that alpha1A-adrenoceptor is a major subtype detectable in binding and RT-PCR studies in rabbit iris, but that the adrenergic contractions of iris dilator muscle are mediated via activation of alpha1-adrenoceptor subtype having low affinity for prazosin and WB4101. PMID- 10455287 TI - Differential blockade of rat alpha3beta4 and alpha7 neuronal nicotinic receptors by omega-conotoxin MVIIC, omega-conotoxin GVIA and diltiazem. AB - Rat alpha3beta4 or alpha7 neuronal nicotinic acetylcholine receptors (AChRs) were expressed in Xenopus laevis oocytes, and the effects of various toxins and non toxin Ca2+ channel blockers studied. Nicotinic AChR currents were elicited by 1 s pulses of dimethylphenylpiperazinium (DMPP, 100 microM) applied at regular intervals. The N/P/Q-type Ca2+ channel blocker omega-conotoxin MVIIC inhibited alpha3beta4 currents with an IC50 of 1.3 microM; the blockade was non-competitive and reversible. The alpha7 currents were unaffected. At 1 microM, omega-conotoxin GVIA (N-type Ca2+ channel blocker) inhibited by 24 and 20% alpha3beta4 and alpha7 currents, respectively. At 1 microM, omega-agatoxin IVA (a P/Q-type Ca2+ channel blocker) did not affect alpha7 currents and inhibited alpha3beta4 currents by only 15%. L-type Ca2+ channel blockers furnidipine, verapamil and, particularly, diltiazem exhibited a preferential blocking activity on alpha3beta4 nicotinic AChRs. The mechanism of alpha3beta4 currents blockade by omega-conotoxins and diltiazem differed in the following aspects: (i) the onset and reversal of the blockade was faster for toxins; (ii) the blockade by the peptides was voltage dependent, while that exerted by diltiazem was not; (iii) diltiazem promoted the inactivation of the current while omega-toxins did not. These data show that, at concentrations currently employed as Ca2+ channel blockers, some of these compounds also inhibit certain subtypes of nicotinic AChR currents. Our data calls for caution when interpreting many of the results obtained in neurons and other cell types, where nicotinic receptor and Ca2+ channels coexist. PMID- 10455288 TI - Pharmacological characterization of Ca2+ entry channels in endothelin-1-induced contraction of rat aorta using LOE 908 and SK&F 96365. AB - We have recently shown that endothelin-1 (ET-1) activates two types of Ca2+ permeable nonselective cation channels (designated NSCC-1 and NSCC-2) and store operated Ca2+ channel (SOCC). These channels can be pharmacologically discriminated using Ca2+ channel blockers such as SK&F 96365 and LOE 908. Here we characterized Ca2+ entry channels involved in ET-1-induced contractions of rat thoracic aortic rings and increases in the intracellular free Ca2+ concentration ([Ca2+]i) of single smooth muscle cells using these blockers. LOE 908 or a blocker of voltage-operated Ca2+ channel nifedipine had no effect on the contractions and increases in [Ca2+]i induced by thapsigargin or ionomycin, whereas SK&F 96365 abolished them. The contractions and increases in [Ca2+]i induced by ET-1 depended on extracellular Ca2+ but were resistant to nifedipine. The responses to lower concentrations (< or =0.1 nM) of ET-1 were abolished by either SK&F 96365 or LOE 908. The responses to higher concentrations (> or = 1 nM) were abolished by SK&F 96365, but were partially resistant to LOE 908. SK&F 96365 inhibited the LOE 908-resistant contractions induced by higher concentrations of ET-1 with IC50 values similar to those for contractions induced by thapsigargin or ionomycin. These results show that the contractions and increases in [Ca2+]i of rat aortic smooth muscles at lower concentrations of ET-1 involve only one Ca2+ entry channel which is sensitive to SK&F 96365 and LOE 908 (NSCC-2), whereas those at higher concentrations of ET-1 involve another Ca2+ entry channel which is sensitive to SK&F 96365 but resistant to LOE 908 (SOCC) in addition to the former channel. PMID- 10455290 TI - Suc-[Glu9,Ala11,15]-endothelin-1 (8-21), IRL 1620, identifies two populations of ET(B) receptors in guinea-pig bronchus. AB - The pharmacological properties of endothelin receptors (ETR) were investigated in guinea-pig bronchus by comparing binding and functional results. In binding assays, both the ET(B) agonists, endothelin-3 (ET-3) and N-suc-[Glu9,Ala11,15]ET 1(8-21) (IRL 1620), and the antagonist, N-cis-2,6-dimethylpiperidinocarbonyl-L gamma-methylleucyl-D- 1-methoxycarbonyltryptophanyl-D-norleucine (BQ 788), showed biphasic inhibition curves of [125I]-endothelin-1 (ET-1) binding to bronchus membranes prepared from intact or epithelium-deprived tissue. IRL 1620 did not completely displace specifically [125I]-ET-1 bound to these tissue preparations. In the presence of the ET(A)-selective antagonist, cyclo(-D-Trp-D-Asp-L-Pro-D-Val L-Leu) (BQ 123, 1 microM), IRL 1620 displacement curves were shallow but a complete inhibition was reached at a concentration of 1 microM. Both curves were better represented by two-site models. In addition, BQ 788 competition curves became monophasic when binding experiments were performed in the presence of 1 microM BQ 123. The non-selective agonist, ET-1, and BQ 123 inhibited [125I]-ET binding to bronchus membranes in dose-dependent fashions with monophasic curves. The contracting activity of IRL 1620 (0.55 nM- 1.6 microM) was tested on multiple ring bronchial preparations pretreated with peptidase and cyclo-oxygenase inhibitors. BQ 788 shifted IRL1620 concentration-response curves to the right while BQ 123 did not influence bronchial responsiveness. In addition, a potentiation of the maximal response to the agonist was observed in BQ 788 treated bronchial rings. This effect was abolished by tissue pretreatment with Nomega-nitro-L-argininemethylester (L-NAME) or epithelium removal but not by pretreatment with atropine or iberiotoxin. Our results demonstrate that guinea pig bronchus contains two populations of ET(B) receptors with different affinities for the ET(B)-selective agonist, IRL 1620. One ET(B) receptor population appears to activate bronchial muscle contraction while another on epithelial cells causes muscle relaxation through the release of nitric oxide (NO). PMID- 10455289 TI - Contribution of cytochrome P450 epoxygenase and hydroxylase pathways to afferent arteriolar autoregulatory responsiveness. AB - Previous studies have demonstrated an important role for the cytochrome P450 (CYT P450) pathway in afferent arteriole autoregulatory responses but the involvement of specific pathways remains unknown. Experiments were performed to determine the role of CYT-P450 epoxygenase and hydroxylase pathways in pressure mediated preglomerular autoregulatory responses. Afferent arteriolar diameter was measured as renal perfusion pressure was increased from 80-160 mmHg. Afferent arteriolar diameter averaged 19+/-2 microm at a renal perfusion pressure of 80 mmHg and decreased by 15+/-2% when pressure was increased to 160 mmHg. Inhibition of the epoxygenase pathway with 6-(2-proparglyloxyphenyl)hexanoic acid (PPOH), enhanced the microvascular response to increasing renal perfusion pressure. In the presence of 50 microM PPOH, afferent arteriolar diameter decreased by 29+/-4% when pressure was increased from 80-160 mmHg. Likewise, the sulphonimide derivative of PPOH, N-methylsulphonyl-6-(2-proparglyloxyphenyl) hexanamide (MS PPOH, 50 microM), enhanced the afferent arteriolar response to increasing renal perfusion pressure. In contrast, the selective CYT-P450 hydroxylase inhibitor, N methylsulphonyl-12,12-dibromododec-11-enamide (DDMS) attenuated the vascular response to increasing renal perfusion pressure. In the pressure of 25 microM DDMS, afferent arteriolar diameter decreased by 4+/-2% when pressure was increased from 80-160 mmHg. These results suggest that CYT-P450 metabolites of the epoxygenase pathway alter afferent arteriolar responsiveness and thereby modify the ability of the preglomerular vasculature to autoregulate renal blood flow. Additionally, these results provide further support to the concept that a metabolite of the hydroxylase pathway is an integral component of the afferent arteriolar response to elevations in perfusion pressure. PMID- 10455291 TI - Endothelin generating pathway through endothelin1-31 in human cultured bronchial smooth muscle cells. AB - The effects of endothelin (ET)-1(1-31) and ET-2(1-31), human chymase products of the corresponding big ETs, on the intracellular free Ca2+ concentration ([Ca2+]i) and [125I]-ET-1 binding were investigated using human cultured bronchial smooth muscle cells (BSMC). ET-1(1-31) (10(-8)M - 3 x 10(-7)M) and ET-2(1-31) (3 x 10( 8)M - 3 x 10(-6) M) caused an increase in [Ca2+]i in a concentration-dependent manner. Big ET-1 (3 x 10(-8)M - 10(-6)M) also caused this increase, but not big ET-2 at concentrations up to 10(-6)M. The [Ca2+]i increase induced by ET-1 was inhibited by both BQ123, an ET(A)-receptor antagonist, and BQ788, an ET(B) receptor antagonist, whereas that induced by ET-3 was inhibited by BQ788 but not by BQ123. Increases in [Ca2+]i caused by ET-1(1-31), big ET-1 and ET-2(1-31) were completely inhibited by 10(-4)M phosphoramidon, a dual neutral endopeptidase (NEP)/endothelin-converting enzyme (ECE) inhibitor, and 10(-5)M thiorphan, a NEP inhibitor. Scatchard plot analyses of the saturation curves of [125I]-ET-1 and [125I]-ET-3 showed that both ET(A)- and ET(B)- receptors at the ratio of 4:1 were expressed on BSMC. ET-1(1-31), big ET-1 and ET-2(1-31) inhibited [125I]-ET-1 binding in a concentration-dependent manner, and these effects were attenuated by treatment with thiorphan. On the other hand, big ET-2 slightly inhibited the binding at a high concentration and this was not affected by thiorphan. These results suggest that ET-1(1-31), big ET-1 and ET-2(1-31) cause an increase in [Ca2+]i by being converted into the corresponding ET-1 and ET-2 by NEP, but this did not occur with big ET-2 in human BSMC. ET-2(1-31), produced by human chymase from big ET-2 might be important for the generation of ET-2 in human bronchial tissue. PMID- 10455292 TI - Presynaptic inhibition preferentially reduces in NMDA receptor-mediated component of transmission in rat midbrain dopamine neurons. AB - We used patch pipettes to record whole-cell currents from single dopamine neurons in slices of rat midbrain. Pharmacological methods were used to isolate EPSCs evoked by focal electrical stimulation. Baclofen was significantly more potent for inhibiting NMDA receptor-mediated EPSCs (IC50=0.24 microM) compared with inhibition of EPSCs mediated by AMPA receptors (IC50=1.72 microM). The increased potency of baclofen for inhibiting the NMDA component persisted in superfusate that contained zero Mg2+ and when postsynaptic K+ conductances were reduced by Cs+ and QX-314. Effects of baclofen on EPSCs were blocked by the GABA(B) receptor antagonist CGP-35348. Adenosine was 20 fold more potent for reducing the NMDA component of transmission (IC50=31 microM) compared with inhibition of AMPA receptor-mediated EPSCs (IC50=654 microM). Effects of adenosine on EPSCs were blocked by the A1 receptor antagonist DPCPX. Both baclofen and adenosine significantly increased the ratio of EPSCs in paired-pulse studies, suggesting presynaptic sites of action. Although adenosine (1 mM) did not reduce currents evoked by exogenous NMDA (10 microM), baclofen (1 microM) reduced NMDA currents by 29%. Neither baclofen nor adenosine altered currents evoked by exogenous AMPA (1 microM). We conclude that adenosine acts at presynaptic A1 receptors to cause a preferential reduction in the NMDA component of synaptic transmission. In contrast, baclofen preferentially reduces NMDA EPSCs by acting at both pre- and postsynaptic GABA(B) receptors. By regulating NMDA receptor function, A1 and GABA(B) receptors may play important roles in regulating the excitability of dopamine neurons. PMID- 10455293 TI - Pharmacological characterization of 5-HT4 receptors mediating relaxation of canine isolated rectum circular smooth muscle. AB - This study aimed to characterize for the first time in vitro 5-HT4 receptors in the canine gastrointestinal tract. For this purpose, we used circular muscle strips of the canine isolated rectum. In the presence of methysergide (60 microM), 5-HT induced relaxation of methacholine (1 microM)-precontracted muscle strips, yielding a monophasic sigmoidal concentration-relaxation curve (pEC50 7.2+/-0.07). Tetrodotoxin (0.3 microM) did not affect the curve to 5-HT, suggesting the inhibitory 5-HT receptor is located on the smooth muscle. Granisetron (0.3 microM) did also not affect the curve to 5-HT, which excludes the 5-HT3 receptor mediating the relaxation to 5-HT. The presence of methysergide rules out the involvement of 5-HT1, 5-HT2 or 5-HT7 receptors. 5-HT, the selective 5-HT4 receptor agonists R076186, prucalopride (R093877) and SDZ HTF-919 and the 5 HT4 receptor agonists cisapride and 5-MeOT relaxed the muscle strips with a rank order of potency R076186 = 5-HT > cisapride > prucalopride > or = SDZ HTF-919 > 5 MeOT. The selective 5-HT4 receptor antagonists GR 125487, RS 39604 and GR 113808 competitively antagonized the relaxations to 5-HT, yielding pK(B) estimates of 9.7, 7.9 and 9.1, respectively. The selective 5-HT4 receptor antagonist SB 204070 shifted the curve to 5-HT rightward and depressed the maximal response (apparent pA2 10.6). GR 113808 (10 nM) produced a parallel rightward shift of the curve to the selective 5-HT4 receptor agonists R076186 (pA2 8.8). It is concluded that 5 HT induces relaxation of the canine rectum circular muscle through stimulation of a single population of smooth muscle 5-HT4 receptors. For the first time, a nonhuman species was shown to exhibit relaxant 5-HT4 receptors in the large intestine. PMID- 10455294 TI - Atriopeptin, sodium azide and cyclic GMP reduce secretion of aqueous humour and inhibit intracellular calcium release in bovine cultured ciliary epithelium. AB - This study examined the involvement of cyclic GMP, protein kinase G and intracellular Ca2+ movements in the modulation of aqueous humour formation. Using the bovine arterially-perfused eye preparation, drug effects on intraocular pressure and aqueous humour formation rate were measured by manometry and fluorescein dilution, respectively. Drug effects on intracellular [Ca2+] were determined by fura-2 fluorescence ratio technique in nontransformed, cultured ciliary epithelium. Intra-arterial injection of atriopeptin (50 pmol) or sodium azide (10 nmol) produced significant reduction in aqueous humour formation (>38%). This was blocked by selective inhibition (KT-5823) of protein kinase G, but not by selective inhibition (KT-5720) of protein kinase A. Reductions of intraocular pressure produced by atriopeptin or azide were almost completely blocked by KT-5823. ATP (100 microM) caused rapid, transient increase in intracellular Ca2+ followed by a slow decline and prolonged plateau. This response showed concentration-dependent inhibition by atriopeptin, azide or 8 bromo cyclic GMP, and this inhibition of the rapid (peak) Ca2+ increase was enhanced by zaprinast (100 microM; phosphodiesterase inhibitor). KT-5823 blocked the suppression of the peak Ca2+ response but not suppression of the plateau. Arterial perfusion of ATP (0.1-100 microM) produced a concentration-dependent decrease in aqueous humour formation. Aqueous humour formation in the bovine eye can be manipulated through cyclic GMP, operating via protein kinase G. Close parallels appear when Ca2+ movements are modified by similar manipulations of cyclic GMP, suggesting that Ca2+ transients may play an important role in aqueous humour formation and that interplay occurs between cyclic GMP and Ca2+. PMID- 10455295 TI - Involvement of ATP-sensitive potassium channels in a model of a delayed vascular hyporeactivity induced by lipopolysaccharide in rats. AB - We have investigated the role of ATP-sensitive potassium (K(ATP)) channels in an experimental model of a delayed phase of vascular hyporeactivity induced by lipopolysaccharide (LPS) in rats. After 24 h, from LPS treatment, in anaesthetized rats the bolus injection of phenylephrine (PE) produced an increase in mean arterial pressure (MAP) significantly (P<0.05) reduced in LPS-treated rats compared to the vehicle-treated rats. This reduction was prevented by pre treatment of rats with glibenclamide (GLB), a selective inhibitor of K(ATP) channels. GLB administration did not affect the MAP in vehicle-treated rats but produced an increase of MAP in rats treated with LPS. Cromakalim (CRK), a selective K(ATP) channel opener, produced a reduction of MAP that was significantly (P<0.05) higher in LPS- than in vehicle-treated rats. In contrast, the hypotension induced by glyceryl trinitrate (GTN) in LPS-treated rats was not distinguishable from that produced in vehicle-treated rats. Experiments in vitro were conducted on aorta rings collected from rats treated with vehicle or LPS 24 h before sacrifice. The concentration-dependent curve to PE was statistically (P<0.005) reduced in aorta rings collected from LPS- compared to vehicle-treated rats. This difference was totally abolished by tetraethylammonium (TEA), a non selective inhibitor of K+ channels. CRK produced a relaxation of PE precontracted aorta rings higher in rings from LPS- than in vehicle-treated rats. GLB inhibited CRK-induced relaxation in both tissues, abolishing the observed differences. In conclusion, our results indicate an involvement of K(ATP) channels to the hyporesponsiveness of vascular tissue after 24 h from a single injection of LPS in rats. We can presume an increase in the activity of K(ATP) channels on vascular smooth muscle cells but we cannot exclude an increase of K(ATP) channel number probably due to the gene expression activation. PMID- 10455296 TI - Spinal effect of a neuropeptide FF analogue on hyperalgesia and morphine-induced analgesia in mononeuropathic and diabetic rats. AB - 1DMe, a neuropeptide FF (NPFF) analogue, has been shown to produce antinociception and to enhance morphine analgesia in rats after intrathecal administration. To determine whether 1DMe could correct hyperalgesia and restore morphine efficacy in mononeuropathic (MN) and diabetic (D) rats we examined the spinal effect of 1DMe in MN and D rats without and after spinal blockade of mu- and delta-opioid receptors with CTOP and naltrindole, respectively. The influence of 1DMe on morphine-induced antinociception was assessed in the two models using isobolographic analysis. Whereas 1DMe intrathecally injected (0.1, 1, 7.5 microg rat(-1)) was ineffective in normal (N) rats, it suppressed mechanical hyperalgesia (decrease in paw pressure-induced vocalisation thresholds) in both MN and D rats. This effect was completely cancelled by CTOP (10 microg rat(-1)) and naltrindole (1 microg rat(-1)) suggesting that it requires the simultaneous availability of mu- and delta-opioid receptors. The combinations of morphine: 1DMe (80.6:19.4% and 99.8:0.2%, in MN and D rats, respectively) followed by isobolographic analysis, showed a superadditive interaction, relative to the antinociceptive effect of single doses, in D rats only. In N rats, the combination of morphine: 1DMe (0.5 mg kg(-1), i.v.: 1 microg rat(-1), i.t., ineffective doses) resulted in a weak short-lasting antinociceptive effect. These results show a different efficacy of 1DMe according to the pain model used, suggesting that the pro-opioid effects of the NPFF in neuropathic pain are only weak, which should contribute to hyperalgesia and to the impaired efficacy of morphine. PMID- 10455297 TI - Evidence for two different P2X-receptors mediating vasoconstriction of Ap5A and Ap6A in the isolated perfused rat kidney. AB - The activation of various P2-receptor subtypes in rat renal vasculature by P1, P5 diadenosine pentaphosphate (ApsA) and P1, P6-diadenosine hexaphosphate (Ap6A) were studied by measuring their effects on perfusion pressure during continuous perfusion in a rat isolated perfused kidney. Permanent perfusion with Ap5A and Ap6A elicited both a transient and sustained vasoconstriction with both vasoconstrictions to be different: the transient vasoconstriction can be elicited with concentrations > or = 10 nM, whereas the sustained vasoconstriction is observed with concentrations > or = 1 nM. ApsA and Ap6A act via the same receptors as alpha,beta-methylene ATP (alpha,beta-meATP). The rank order of potency for transient vasconstriction was alpha,beta-meATP = ApsA>Ap6A>B,gamma meATP, and for sustained vasoconstriction alpha,beta-meATP = Ap5A > beta,gamma meATP > or = Ap6A. Suramin, a non-selective P2-receptor antagonist, and pyridoxal phosphate-6-azophenyl-2;4-disulphonic acid (PPADS) a highly selective P2X receptor antagonist antagonized both the transient and the sustained vasoconstriction. Taken together the results of the agonist profile of Ap5A and Ap6A and comparing its findings to literature it can be demonstrated that the transient but not the sustained vasoconstriction is mediated via the P2X1 receptor which is present in rat renal vasculature. It is demonstrated that the agonist profile of the sustained vasoconstriction induced by ApsA and Ap6A does not fit to any currently known P2X- or P2Y-receptor subtype. We conclude a yet unidentified P2X-receptor or chimeric P2X-receptor may contribute to the effects on rat renal vasculature produced by Ap5A and Ap6A and which may play an important role in glomerular perfusion pressure and blood pressure control. PMID- 10455298 TI - Calcium antagonistic properties of the cyclooxygenase-2 inhibitor nimesulide in human myometrial myocytes. AB - The non-steroidal anti-inflammatory drug nimesulide is a selective inhibitor of cyclooxygenase-2 which relaxes spontaneously contracting human myometrium in vivo and is potentially a useful tocolytic drug. Part of the relaxant action of nimesulide may be via block of myometrial Ca2+ channels. Here, we describe the Ca2+ channel blocking properties of nimesulide in freshly dispersed human term pregnant myometrial smooth muscle cells (HMSMCs). Both L- and T-components of the whole cell Ca2+ channel current were inhibited by 100 microM nimesulide (38+/-3 and 35+/-1% block, respectively). At physiological pH inside and outside the cell (pHo/pHi = 7.4/7.2), this block did not depend on the holding or test potential, although a degree of use-dependence was observed during high frequency stimulation at a higher concentration of drug (300 microM). At pHo/pHi = 6.8, under which condition the concentration of the non-ionized form of the drug is increased 3 fold compared to pH 7.4, nimesulide blocked the L-type current more potently (58+/-3% inhibition at 100 microM, P<0.01) compared to physiological pH. Nimesulide caused a 7 mV leftward shift in the availability curve of the current at pH 6.8, suggesting that the affinity of the drug for the inactivated channel is approximately 4 fold higher than its affinity for the closed channel. We speculate that acidification and depolarization of the myometrium during the intense and prolonged contractions of labour might increase the potency of nimesulide as a Ca2+ channel antagonist, promoting its action as a tocolytic agent. PMID- 10455300 TI - Pharmacological characterization of nicotine-induced acetylcholine release in the rat hippocampus in vivo: evidence for a permissive dopamine synapse. AB - In this study, the mechanism of nicotine-induced hippocampal acetylcholine (ACh) release in awake, freely moving rats was examined using in vivo microdialysis. Systemic administration of nicotine (0.4 mg kg(-1), s.c.) increased the levels of ACh in hippocampal dialysates. The nicotine-induced hippocampal ACh release was sensitive to the pretreatment of neuronal nicotinic acetylcholine receptor (nAChR) antagonists mecamylamine (3.0 mg kg(-1), s.c.) and dihydro-beta erythrodine (DHbetaE; 4.0 mg kg(-1), s.c.) as well as systemic administration of the dopamine (DA) D1 receptor antagonist SCH-23390 (R-(+)-7-chloro-8-hydroxy-3 methyl-1-phenyl-2,3,4,5-tetrahydro-1H-benzaz epine; 0.3 mg kg(-1), s.c.). Local perfusion of mecamylamine (100 microM), DHbetaE (100 microM) or SCH-23390 (10 microM) through microdialysis probe did not increase basal hippocampal ACh release. Hippocampal ACh release elicited by systemic administration of nicotine (0.4 mg kg(-1), s.c.) was antagonized by local perfusion of SCH-23390 (10 microM), but not by MEC (100 microM) or DHbetaE (100 microM). Direct perfusion of nicotine (1 mM, but not 0.1 mM) increased hippocampal ACh levels; however, this effect was relatively insensitive to blockade by co-perfusion of either mecamylamine (100 microM) or SCH-23390 (10 microM). These results suggest that nicotine-induced hippocampal ACh release occurs by two distinct mechanisms: (1) activation of nAChRs outside the hippocampus leading to DA release and subsequent ACh release involving a permissive DA synapse, and (2) direct action of nicotine within the hippocampus leading to ACh release via non-DA-ergic mechanism. PMID- 10455301 TI - Evidence that nitric oxide acts as an inhibitory neurotransmitter supplying taenia from the guinea-pig caecum. AB - Nitric oxide synthase-containing nerve fibres are abundant within taenia of the guinea-pig caecum, but there is little previous evidence supporting a direct role for nitric oxide (NO) in responses to enteric inhibitory nerve stimulation. In this study we have attempted to identify an NO-dependent component of inhibitory transmission in isolated taenia coli. Isometric tension was recorded in the presence of atropine and guanethidine (both 1 microM). Tone was raised with histamine (1 microM), and intrinsic inhibitory neurons stimulated using either a nicotinic agonist (1,1-dimethyl-4-phenylpiperazinium iodide; DMPP) or electrical field stimulation (EFS). DMPP (1-100 microM) produced concentration-dependent biphasic relaxations, comprising an initial peak relaxation followed by a sustained relaxation. Responses to DMPP were antagonized by tetrodotoxin (1 microM) or apamin (0.3 microM) and abolished by hexamethonium (300 microM). L nitro-arginine (L-NOARG; 100 microM) and oxyhaemoglobin (2%) both significantly reduced sustained relaxations produced by DMPP. EFS (5 Hz, 30 s) also produced biphasic relaxations. Both L-NOARG and an inhibitor of soluble guanylate cyclase (ODQ, 1-10 microM) reduced the sustained component of EFS responses. Two NO donors, sodium nitroprusside (SNP) and diethylenetriamine-nitric oxide adduct (DENO), produced concentration-dependent relaxations. Responses to SNP and DENO were antagonized by ODQ (1 microM) and by apamin (0.3 mM). These results suggest that NO contributes directly to a component of inhibitory transmission in guinea pig taenia coli. The actions of NO appear to be mediated via cyclic GMP synthesis, and may involve activation of small conductance calcium activated K+ channels. A role for NO is most evident during sustained relaxations evoked by longer stimulus trains or chemical stimulation of intrinsic neurons. PMID- 10455299 TI - Different effect of simvastatin and atorvastatin on key enzymes involved in VLDL synthesis and catabolism in high fat/cholesterol fed rabbits. AB - The effects of atorvastatin (3 mg kg(-1)) and simvastatin (3 mg kg(-1)) on hepatic enzyme activities involved in very low density lipoprotein metabolism were studied in coconut oil/cholesterol fed rabbits. Plasma cholesterol and triglyceride levels increased 19 and 4 fold, respectively, after 7 weeks of feeding. Treatment with statins during the last 4 weeks of feeding abolished the progression of hypercholesterolaemia and reduced plasma triglyceride levels. 3 Hydroxy-3-methyl-glutaryl Coenzyme A reductase, acylcoenzyme A:cholesterol acyltransferase, phosphatidate phosphohydrolase and diacylglycerol acyltransferase activities were not affected by drug treatment. Accordingly, hepatic free cholesterol, cholesteryl ester and triglyceride content were not modified. Simvastatin treatment caused an increase (72%) in lipoprotein lipase activity without affecting hepatic lipase activity. Atorvastatin caused a reduction in hepatic phospholipid content and a compensatory increase in CTP:phosphocholine cytidylyl transferase activity. The results presented in this study suggest that, besides the inhibitory effect on 3-hydroxy-3-methyl-glutaryl Coenzyme A reductase, simvastatin and atorvastatin may have additional effects that contribute to their triglyceride-lowering ability. PMID- 10455303 TI - Central control of blood pressure by nitrergic mechanisms in organum vasculosum laminae terminalis of rat brain. AB - Experiments were carried out to explore the possible role played by the nitric oxide (NO) system in the organum vasculosum laminae terminalis (OVLT) of rat brain in arterial pressure regulation. Intracerebroventricular (ICV) or intra OVLT administration of NO donors such as hydroxylamine, sodium nitro-prusside or s-nitro-acetylpenicillamine caused an up to 55 mmHg decrease in blood pressure (BP) but an increase in NO release (measured by porphyrin/nafion coated carbon fibre electrodes in combination with voltammetry) in the OVLT. In contrast, ICV or intra-OVLT administration of N(G)-nitro-L-arginine methyl ester (L-NAME; a constitutive NO synthase inhibitor) caused an up to 45 mmHg increase in BP but a fall in NO release in the OVLT. Compared with the BP responses induced by ICV injection of NO donors or NO synthase inhibitors, the OVLT route of injection required a much lower dose of NO donors or NO synthase inhibitors to produce a similar BP effect. The depressor effects induced by ICV or intra-OVLT administration of NO donors were attenuated by pretreatment with intra-OVLT injection of methylene blue (an inhibitor of guanylate cyclase), haemoglobin (a NO scavenger), L-NAME or spinal transection. On the other hand, the L-NAME induced pressor effects were attenuated by pretreatment with intra-OVLT injection of L-arginine or spinal transection. The data suggest that activation of cyclic GMP-dependent NO synthase in the OVLT of rat brain causes cyclic GMP-dependent decreases in arterial pressure via inhibiting the sympathetic efferent activity. PMID- 10455302 TI - The ether lipid ET-18-OCH3 increases cytosolic Ca2+ concentrations in Madin Darby canine kidney cells. AB - The effect of the ether lipid 1-O-octadecyl-2-O-methyl-sn-glycero-3 phosphorylcholine (ET-18-OCH3) on the intracellular free Ca2+ concentration ([Ca2+]i) in Madin Darby canine kidney (MDCK) cells was studied using fura-2 as the Ca2+ probe. In Ca2+ medium, ET-18-OCH3 induced a significant rise in [Ca2+]i at concentrations between 10-100 microM with a concentration-dependent delay of 45-175 s. The [Ca2+]i signal was composed of a gradual rise and a sustained plateau. In Ca2+-free medium, ET-18-OCH3 (10-100 microM) induced a Ca2+ release from internal Ca2+ stores with a concentration-dependent delay of 45-175 s. This discharge of internal Ca2+ triggered capacitative Ca2+ entry in a concentration dependent manner. This capacitative Ca2+ entry was not inhibited by econazole (25 microM), 1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole hydrochloride (SKF96365; 50 microM), nifedipine (10 microM), verapamil (10 microM), diltiazem (10 microM) and cadmium (0.5 microM). Methyl 2 (phenylthio)ethyl-1,4-dihydro-2,4,6-trimethylpyridine-3,5-dicarboxylat e (PCA 4248), a platelet-activating factor (PAF) receptor antagonist, inhibited 25 microM ET-18-OCH3-induced [Ca2+]i rise in a concentration-dependent manner between 1-20 microM, with 20 microM exerting a complete block. The [Ca2+]i rise induced by ET-18-OCH3 (25 microM) was not altered when the production of inositol 1,4,5-trisphosphate (IP3) was suppressed by the phospholipase C inhibitor U73122 (2 microM), but was partly inhibited by the phospholipase D inhibitor propranolol (0.1 mM) or the phospholipase A2 inhibitor aristolochic acid (20-40 microM). In Ca2+-free medium, pretreatment with 25 microM ET-18-OCH3 completely depleted the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin-sensitive Ca2+ store. In contrast, pretreatment with thapsigargin abolished 0.1 mM ATP-induced [Ca2+]i rise without altering the ET-18-OCH3-induced [Ca2+]i rise. This suggests that ET 18-OCH3 depleted thapsigargin-sensitive Ca2+ stores and also released Ca2+ from thapsigargin-insensitive stores. The thapsigargin-insensitive stores involve mitochondria because the mitochondria uncoupler carbonylcyanide m chlorophenylhydrazone (CCCP; 2 microM) induced a release of mitochondrial Ca2+ which was abolished by pretreatment with 25 microM ET-18-OCH3. ET-18-OCH3 (25 microM) induced a significant Mn2+ quench of fura-2 fluorescence at 360 nm excitation wavelength confirming that ET-18-OCH3 induced capacitative Ca2+ entry. La3+ (0.1 mM) or Gd3+ (50 microM) abolished the ET-18-OCH3-induced Mn2+ quench and [Ca2+]i rise. Our data imply that ET-18-OCH3 induced a [Ca2+]i rise in MDCK cells by activating PAF receptors leading to an internal Ca2+ release followed by capacitative Ca2+ entry. Phospholipase D and phospholipase A2, but not phospholipase C, might be involved in mediating the capacitative Ca2+ entry. La3+ abolished the ET-18-OCH3-induced [Ca2+]i rise presumably by inhibiting PAF receptors. PMID- 10455304 TI - An inhibitor of poly (ADP-ribose) synthetase activity reduces contractile dysfunction and preserves high energy phosphate levels during reperfusion of the ischaemic rat heart. AB - The cardioprotective properties of inhibition of poly (ADP-ribose) synthetase (PARS) were investigated in the isolated perfused heart of the rat. Hearts were perfused in the Langendorff mode and subjected to 23 min total global ischaemia and reperfused for 60 min. Left ventricular function was assessed by means of an intra-ventricular balloon. High energy phosphates were measured by 31P-NMR spectroscopy. Intracellular levels of NAD were measured by capillary electrophoresis of perchloric acid extracts of hearts at the end of reperfusion. Reperfusion in the presence of the PARS inhibitor 1,5 didroxyisoquinoline (ISO, 100 microM) attenuated the mechanical dysfunction observed following 1 h of reperfusion; 27+/-13 and 65+/-8% recovery of preischaemic rate pressure product for control and 100 microM ISO, respectively. This cardioprotection was accompanied by a preservation of intracellular high-energy phosphates during reperfusion; 38+/-2 vs 58+/-4% (P<0.05) of preischaemic levels of phosphocreatine (PCr) for control and 100 microM ISO respectively and 23+/-1 vs 31+/-3% (P < 0.05) of preischaemic levels of ATP for control and 100 microM ISO respectively. Cellular levels of NAD were higher in ISO treated hearts at the end of reperfusion; 2.56+/-0.45 vs 4.76+/-1.12 micromoles g(-1) dry weight (P<0.05) for control and ISO treated. These results demonstrate that the cardioprotection afforded by inhibition of PARS activity with ISO is accompanied by a preservation of high-energy phosphates and cellular NAD levels and suggest that the mechanism responsible for this cardioprotection may involve prevention of intracellular ATP depletion. PMID- 10455305 TI - Alternative splicing generates two isoforms of the beta3-adrenoceptor which are differentially expressed in mouse tissues. AB - The beta3-adrenoceptor (AR) differs from the beta1-AR and beta2-ARs in having introns within and downstream of the coding block. This study demonstrates two splice variants of the mouse beta3-AR which differ within the coding region. Reverse transcription/polymerase chain reaction with intron-spanning primers was used to demonstrate the splice variant of the mouse beta3-adrenoceptor. The novel beta3b-AR has 17 amino acids encoded by exon 2 (SSLLREPRHLYTCLGYP) which differ from the 13 in the known beta3a-AR (RFDGYEGARPFPT). Beta3b-AR mRNA is differentially expressed in mouse tissues, with levels relative to beta3a-AR mRNA highest in hypothalamus, cortex and white adipose tissue, and lower in ileum smooth muscle and brown adipose tissue. PMID- 10455306 TI - GABA(B), opioid and alpha2 receptor inhibition of calcium channels in acutely dissociated locus coeruleus neurones. AB - 1. The effects of GABA(B), opioid and alpha2 receptor activation on different subtypes of calcium channels in acutely-dissociated rat locus coeruleus (LC) neurones were investigated using whole-cell patch clamping. 2. Barium currents through calcium channels could be fractionated into four classes: L-type (nimodipine-sensitive), N-type (omega-conotoxin GVIA-sensitive), P/Q-type (omega agatoxin IVA-sensitive) and R-type (remaining in the presence of all three blockers). The percentage of each was, respectively, 25+/-2, 34+/-1, 28+/-3 and 12+/-1% (mean+/-s.e.mean, n=4). 3. The GABA(B) receptor agonist, baclofen, and the opioid receptor agonist, enkephalin, partially inhibited the total barium current in a concentration-dependent manner with EC50 values of 2 and 0.3 microm , respectively. Maximal inhibition was 17+/-1% (n=38) for baclofen and 30+/-2% (n=20) for enkephalin. The alpha2-adrenoceptor agonist, UK14304 (10 microM), also inhibited barium current in these neurones (28+/-2%, n=11). The agonists did not shift the current-voltage relationship along the voltage axis. 4. Maximal baclofen inhibition of different calcium channel subtypes was 9+/-7% (L-type, n=4), 11+/-8% (N-type, n=4), 26+/-6% (P/Q-type, n=4), and 6+/-5% (R-type, n=5). The corresponding values for enkephalin inhibition were 5+/-9% (L-type), 30+/-11% (N-type), 37+/-9% (P/Q-type), and 17+/-8% (R-type). 5. In the presence of a saturating concentration of enkephalin, baclofen produced additional inhibition of the barium current. In contrast, in the presence of a saturating concentration of enkephalin, UK14304 produced no further inhibition of the barium current. 6. These results indicate that neuromodulation of calcium channels in LC neurones involves a complex pattern of overlapping and distinct second messenger pathways. Regulation of LC neuronal firing activity by the modulation of calcium channels may be important for LC-mediated behaviour such as alertness and vigilance. PMID- 10455307 TI - Involvement of thromboxane A2 in the endothelium-dependent contractions induced by myricetin in rat isolated aorta. AB - 1. The present study was undertaken to analyse the mechanism of the contractile response induced by the bioflavonoid myricetin in isolated rat aortic rings. 2. Myricetin induced endothelium-dependent contractile responses (maximal value=21+/ 2% of the response induced by 80 mM KCl and pD2=5.12+/-0.03). This effect developed slowly, reached a peak within 6 min and then declined progressively. 3. Myricetin-induced contractions were almost abolished by the phospholipase A2 (PLA2) inhibitor, quinacrine (10 microM), the cyclo-oxygenase inhibitor, indomethacin (10 microM), the thromboxane synthase inhibitor, dazoxiben (100 microM), the putative thromboxane A2 (TXA2)/prostaglandin endoperoxide receptor antagonist, ifetroban (3 microM). These contractions were abolished in Ca2+-free medium but were not affected by the Ca2+ channel blocker verapamil (10 microM). 4. In cultured bovine endothelial cells (BAEC), myricetin (50 microM) produced an increase in cytosolic free calcium ([Ca2+]i) which peaked within 1 min and remained sustained for 6 min, as determined by the fluorescent probe fura 2. This rise in [Ca2+]i was abolished after removal of extracellular Ca2+ in the medium. 5. Myricetin (50 microM) significantly increased TXB2 production both in aortic rings with and without endothelium and in BAEC. These increases were abolished both by Ca2+-free media and by indomethacin. 6. Taken together, these results suggests that myricetin stimulates Ca2+ influx and subsequently triggers the activation of the PLA2 and cyclo-oxygenase pathways releasing TXA2 from the endothelium to contract rat aortic rings. The latter response occurs via the activation of Tp receptors on vascular smooth muscle cells. PMID- 10455308 TI - Pacing-induced delayed protection against arrhythmias is attenuated by aminoguanidine, an inhibitor of nitric oxide synthase. AB - 1. Cardiac pacing, in anaesthetized dogs, protects against ischaemia and reperfusion-induced ventricular arrhythmias when this is initiated 24 h after the pacing stimulus. Now we have examined whether this delayed cardioprotection afforded by cardiac pacing is mediated through nitric oxide. 2. Twenty-two dogs were paced (4 x 5 min periods at 220 beats min(-1)) by way of the right ventricle, 24 h prior to a 25 min period of coronary artery occlusion. Nine of these dogs were given the inhibitor of induced nitric oxide synthase, aminoguanidine (50 mg kg(-1) i.v.), 0.5 h prior to coronary artery occlusion. Sham-operated non-paced dogs with and without aminoguanidine treatment served as controls. 3. Pacing markedly (P<0. 05) reduced arrhythmia severity (ventricular fibrillation, VF, during occlusion 15%; survival from the combined ischaemia reperfusion insult 62%) compared to control, sham-operated, unpaced dogs (VF during occlusion 58%; survival 17%). This protection was attenuated by the administration of aminoguanidine prior to coronary artery occlusion (survival from the combined ischaemia-reperfusion insult 11%, which was significantly (P<0.05) less than in the paced dogs not given aminoguanidine and similar to the controls). Aminoguanidine had no significant effects on coronary artery occlusion when given to dogs that had not been paced. In the dose used aminoguanadine transiently elevated systemic arterial pressure by a mean of 20 mmHg and reduced heart rate by a mean of 22 beats min(-1). 4. These results suggest that nitric oxide, probably derived from induced nitric oxide synthase, contributes significantly to the delayed cardioprotection afforded by cardiac pacing. PMID- 10455309 TI - Characterization of the muscarinic receptor in isolated uterus of sham operated and ovariectomized rats. AB - 1. The pharmacological characteristics of muscarinic receptors in rat isolated uterus were studied in ovariectomized (ov.) and sham operated (sh.) animals. 2. Competition radioligand binding studies, using uterine membranes and [3H]-NMS, were undertaken with several muscarinic receptor antagonists. Most of the antagonists indicated a one-site fit with apparent affinity estimates (pKi) unchanged by ovariectomy. The selective M2 antagonist, tripitramine revealed high (representing 33+/-8 and 38+/-2%) and low (67+/-8 and 62+/-2%) affinity binding sites in both sh. and ov. rat uterus, respectively. These sites likely represented muscarinic M2 and M3 receptors and the proportions were not significantly different in the two conditions. 3. Carbachol induced concentration dependent contractions which were surmountably antagonized by several muscarinic receptor antagonists (pKB, sh.; ov.): zamifenacin (9.19; 9.18), p-F-HHSiD (8. 50; 9.06), tripitramine (7.23; 7.54), himbacine (7.21; 7.41), methoctramine (6.79; 7.49), pirenzepine (6.48; 7.21), AF DX 116 (6. 26; 6.61), MTx 3 (<7.00; <7.00) and PD 102807 (<7.00; <7.00). 4. The apparent affinity values obtained in functional studies using the uteri from both sh. and ov. animals correlated most closely with values reported at human recombinant muscarinic M3 receptors. This suggests that the muscarinic M3 receptor mediates contraction under both conditions. 5. Radioligand binding experiments indicate the presence of M2 receptors, in addition to M3 receptors, which probably explains the discrepancies between functional and binding affinities. These data further suggest that the pharmacological profile and proportions of the two populations of muscarinic receptors are unaffected by ovariectomy. PMID- 10455310 TI - Effects of exercise training on responsiveness of the mesenteric arterial bed to phenylephrine and KCl in male rats. AB - 1. We aimed to determine whether there are any changes in responsiveness of the mesenteric arterial beds to phenylephrine (Phe) and KCl in exercise-trained rats, and whether vascular endothelium and/or vascular smooth muscle play a role in these changes. 2. Adult male rats were subjected to a swimming schedule every day for 28-33 days. Studies were performed in vitro using Krebs perfused mesenteric arterial beds. 3. Maximum perfusion pressure responses to KCl and Phe of the mesenteric arterial beds from exercise-trained rats were significantly lower than those from sedentary controls. However, these differences disappeared after blocking the nitric oxide synthase by NG-nitro-L-arginine (L-NOARG). 4. 3-[(3 cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS, 3 mg ml(-1), 2 min infusion) caused a significant increase in maximum perfusion pressure responses to KCl to the same extent in both exercise-trained and sedentary control rats. CHAPS caused about a 4.5 fold leftward shift of the curve with no change in maximum response to Phe for the mesenteric arterial beds from sedentary control rats, but not for those obtained from exercise-trained rats. However, these differences were abolished in the presence of L-NOARG. 5. Indomethacin did not alter the dose-response curves to KCl or Phe in either swimming or control groups. 6. These results suggest that there was a lower vascular responsiveness to KCl and Phe in exercise-trained rats at rest. The decrease in reactivities to KCl or decrease in sensitivity to Phe after having endothelium impairment by CHAPS of the mesenteric arterial beds of exercise-trained rats were due to an increase in both spontaneous release and upregulation of phenylephrine-stimulated release of nitric oxide from both the vascular endothelium and the vascular smooth muscle cells, and may not be a consequence of an increase in vasodilator prostaglandins by the vascular bed. PMID- 10455311 TI - Anisatin modulation of the gamma-aminobutyric acid receptor-channel in rat dorsal root ganglion neurons. AB - 1. Anisatin, a toxic, insecticidally active component of Sikimi plant, is known to act on the GABA system. In order to elucidate the mechanism of anisatin interaction with the GABA system, whole-cell and single-channel patch clamp experiments were performed with rat dorsal root ganglion neurons in primary culture. 2. Repeated co-applications of GABA and anisatin suppressed GABA-induced whole-cell currents with an EC50 of 1.10 microM. No recovery of currents was observed after washout with anisatin-free solution. 3. However, pre-application of anisatin through the bath had no effect on GABA-induced currents. The decay phase of currents was accelerated by anisatin. These results indicate that anisatin suppression of GABA-induced currents requires opening of the channels and is use-dependent. 4. Anisatin suppression of GABA-induced currents was not voltage dependent. 5. Picrotoxinin attenuated anisatin suppression of GABA induced currents. [3H]-EBOB binding to rat brain membranes was competitively inhibited by anisatin. These data indicated that anisatin bound to the picrotoxinin site. 6. At the single-channel level, anisatin did not alter the open time but prolonged the closed time. The burst duration was reduced and channel openings per burst were decreased indicating that anisatin decreased the probability of openings. PMID- 10455312 TI - Maitotoxin-induced nerve growth factor production accompanied by the activation of a voltage-insensitive Ca2+ channel in C6-BU-1 glioma cells. AB - 1. The aim of the present study was to determine the effects of maitotoxin on nerve growth factor production and the Ca2+ influx in clonal rat glioma cells (C6 BU-1). 2. Maitotoxin (1 - 10 ng ml-1) induced a profound increase in 45Ca2+ influx in an extracellular Ca2+-dependent manner. However, high KCl had no effect at all. These effects were supported by the results from the analysis of intracellular Ca2+ concentration using fura 2. 3. The maitotoxin-induced 45Ca2+ influx was inhibited by inorganic Ca2+ antagonists, such as Mg2+, Mn2+ and Co2+. The inhibitory effect of Co2+ was antagonized by increasing the extracellular Ca2+ concentrations. 4. Maitotoxin (3 ng ml-1) as well as A-23187 (1microM) and dibutyryl cyclic AMP (0.5 mM) caused an acceleration of nerve growth factor (NGF) production in C6-BU-1 cells, as determined by NGF enzyme immunoassay. 5. Reverse transcription polymerase chain reaction (RT - PCR) analysis showed that maitotoxin (10 ng ml-1) enhanced the expression of NGF mRNA, which was abolished by the removal of extracellular Ca2+. A-23187 also accelerated its expression. 6. These results suggest that maitotoxin activates a voltage-insensitive Ca2+ channel and accelerates NGF production mediated through a Ca2+ signalling pathway in C6-BU-1 glioma cells. PMID- 10455313 TI - Repeated challenge with dinitrobenzene sulphonic acid in dinitrofluorobenzene sensitized mice results in vascular hyperpermeability in the trachea: a role for tachykinins. AB - 1. This study investigates the role of tachykinins in a repeated challenge with dinitrobenzene sulphonic acid (DNS) on the tracheal vascular permeability in dinitrofluorobenzene (DNFB)-sensitized mice. 2. DNFB-contact sensitization was followed by an intranasal (i.n.) challenge with DNS. A second challenge with DNS was administered 24 h after the first challenge. To assess changes in tracheal vascular permeability, Evans blue dye accumulation in tracheal tissue was measured. 3. A repeated challenge with DNS in DNFB-sensitized mice led to a 2.8 fold increase in tracheal vascular permeability when compared to DNFB-sensitized and vehicle-challenged mice or a 2.5 fold increase when compared to DNFB sensitized single DNS-challenged mice (P<0.001, ANOVA). 4. RP67580 (10-9 mol mouse-1 i.v.) reduced the increased tracheal vascular permeability induced by a second exposure to DNS in DNFB-sensitized mice completely when injected 15 min before the second challenge (P<0.001, ANOVA). 5. The increased tracheal vascular permeability response induced by the second exposure to DNS could be mimicked with i.n. application of capsaicin (10-10 mol mouse-1) or substance P (SP) (10-12 mol mouse-1) to DNFB-sensitized and single DNS-challenged mice. 6. These results suggest that both tachykinin NK1 receptors and sensory nerves are involved in the development of vascular hyperpermeability changes found in the trachea of DNFB sensitized mice after a repeated DNS-challenge. PMID- 10455314 TI - A771726, the active metabolite of leflunomide, directly inhibits the activity of cyclo-oxygenase-2 in vitro and in vivo in a substrate-sensitive manner. AB - 1. The immunosuppressive and anti-inflammatory drug leflunomide has several sites of action, although its precise mode of action is unknown. 2. Here we show in vitro and in vivo that leflunomide and/or its active metabolite A771726, inhibit the activity of cyclo-oxygenase (COX) at doses below those that affect protein expression. 3. In J774.2 macrophages treated with endotoxin for 24 h to induce COX-2 and iNOS, leflunomide and A771726 inhibited more potently the accumulation of PGE2 (A771726, IC50 3.5 microg ml-1) than of NO2 (A771726, IC50 380 microg ml 1). At high concentrations (>300 microg ml-1) A771726 also exhibited the expression of COX-2 and iNOS proteins. 4. In A549 cells treated for 24 h with interleukin-1beta, to induce COX-2, A771726 potently inhibited PGE2 synthesis (IC50 0.13 microg ml-1). In the same cells, A771726 was notably less active (IC50, 52 microg ml-1) at inhibiting the formation of PGE2 stimulated by exposure to 30 microM arachidonic acid. 5. In a human whole blood assay, measuring the accumulation of TxB2 in response to calcium ionophore as a measure of COX-1 activity and in response to incubation with bacterial endotoxin as a measure of COX-2 activity, leflunomide inhibited COX-1 and COX-2 with IC50 values of 31 and 185 microg ml-1; for A771726 the corresponding values were 40 and 69 microg ml-1. 6. Pre-treatment of rats with leflunomide or A771726 (10 mg kg-1, i.p.) inhibited the plasma accumulation of 6-keto-PGF1alpha but not NO2/NO3 following infusion of endotoxin. Injection of a bolus of arachidonic acid following 6 h infusion of endotoxin caused a marked acute rise in plasma 6-keto-PGF1alpha which was inhibited only by higher doses of A771726 (50 mg kg-1, i.p.). 7. In conclusion, leflunomide via A771726 can directly inhibit the activity of COX, an effect that appears blunted both by increases in substrate supply and possibly by plasma binding. Only at much higher drug levels does leflunomide and/or A771726 inhibit the induction of COX-2 or iNOS proteins. PMID- 10455315 TI - Evidence that the ATP-induced increase in vasomotion of guinea-pig mesenteric lymphatics involves an endothelium-dependent release of thromboxane A2. AB - 1. Experiments were made to investigate mechanisms by which adenosine 5' trisphosphate (ATP) enhanced vasomotion in mesenteric lymphatic vessels isolated from young guinea-pigs. 2. ATP (10-8 - 10-3 M) caused a concentration-dependent increase of perfusion-induced vasomotion with the endothelium mediating a fundamental role at low ATP concentrations (10-8 - 10-6 M). 3. The response to 10 6 M ATP showed tachyphylaxis when applied at intervals of 10 min but not at intervals of 20 or 30 min. 4. Suramin (10-4 M) or reactive blue 2 (3x10-5 M) but not PPADS (3x10-5 M) abolished the excitatory response to 10-6 M ATP confirming an involvement of P2 purinoceptors. 5. The excitatory response to 10-6 M ATP was abolished by treatment with either pertussis toxin (100 ng ml-1), antiflammin-1 (10-9 M), indomethacin (3x10-6 M) or SQ29548 (3x10-7 M), inhibitors of specific G proteins, phospholipase A2, cyclo-oxygenase and thromboxane A2 receptors respectively. 6. ATP simultaneously induced a suramin-sensitive inhibitory response, which was normally masked by the excitatory response. ATP-induced inhibition was mediated by endothelium-derived nitric oxide (EDNO) as the response was abolished by NG-nitro-L-arginine (L-NOARG; 10-4 M), an inhibitor of nitric oxide synthase. 7. We conclude that ATP modulates lymphatic vasomotion by endothelium-dependent and endothelium-independent mechanisms. One of these is a dominant excitation caused through endothelial P2 purinoceptors which because of an involvement of a pertussis toxin sensitive G-protein may be of the P2Y receptor subtype. Their stimulation increases synthesis of phospholipase A2 and production of thromboxane A2, an arachidonic acid metabolite which acts as an endothelium-derived excitatory factor. PMID- 10455316 TI - Synthesis of nitric oxide in the dorsal motor nucleus of the vagus mediates the inhibition of gastric acid secretion by central bombesin. AB - 1. Central administration of bombesin inhibits gastric acid production independently of the centrally or peripherally-acting stimuli employed. This study evaluates the role and location of the cerebral nitric oxide (NO) implicated in the inhibitory effect of central bombesin on in vivo rat gastric acid secretion, as induced by distension with 15 cm H2O, insulin (0.75 u.i. kg-1 i.p.) TRH (1.2 microg kg-1, i.c.) or pentagastrin (100 microg kg-1, i.p.). 2. The acid-inhibitory effect of i.c. bombesin (40 ng kg-1) was prevented by prior administration of L-NAME (80 microg kg-1) in the dorsal motor nucleus of the vagus (DMN). This dose of L-NAME when administered into the nucleus of the tractus solitarious (NTS) did not influence the effects of bombesin. Administration of L-arginine (400 microg kg-1) into the DMN restored the acid inhibitory effect of i.c. bombesin in animals treated with L-NAME. 3. Microinjection of bombesin (12 ng kg-1) into the paraventricular nucleus of the hypothalamus (PvN) inhibits acid secretion stimulated by pentagastrin. This inhibitory effect was prevented by a previous injection of L-NAME (80 microg kg 1) into the DMN. 4. The release of NO in the DMN following i.c. administration of bombesin was confirmed by in vivo electrochemical detection. 5. Administration by microdialysis in the DMN of the NO-donor SNAP (25 mM in 1.5 microl min-1) into the DMN inhibits pentagastrin-stimulated gastric acid secretion. 6. The present study suggests that nNOS-containing neurons in the DMN have an inhibitory role in the control of gastric acid responses. PMID- 10455317 TI - Influence of CGRP (8-37), but not adrenomedullin (22-52), on the haemodynamic responses to lipopolysaccharide in conscious rats. AB - 1. The functional involvement of the vasodilator peptides, adrenomedullin (ADM) and calcitonin gene-related peptide (CGRP), in the haemodynamic sequelae of continuous infusion of lipopolysaccharide (LPS) was assessed in conscious, male, Long Evans rats, by the use of peptide antagonists. 2. It was demonstrated that ADM (22-52) at a dose of 500 nmol kg-1 h-1 caused significant inhibition of the effects of ADM (1 nmol kg-1), without affecting responses to CGRP (0.1 or 1 nmol kg-1). 3. Even when the regional vasodilator responses to LPS infusion were enhanced (by pre-treatment with dexamethasone and the endothelin antagonist, SB 209670, or by pretreatment with SB 209670 and the AT1-receptor antagonist, losartan), ADM (22-52) had no significant cardiovascular effects. In contrast, the CGRP1-receptor antagonist, CGRP (8-37), caused small, but significant, inhibitions of the hypotensive and renal and mesenteric vasodilator effects of LPS, but only 6 h after onset of infusion in the presence of dexamethasone and SB 209670. 4. The results indicate that, in this model of endotoxaemia, the marked regional vasodilatations seen in the presence of dexamethasone and SB 209670 do not involve ADM, but do involve CGRP, albeit only to a small extent. PMID- 10455318 TI - Analysis of the atypical characteristics of adenosine receptors mediating negative inotropic and chronotropic responses of guinea-pig isolated atria and papillary muscles. AB - 1. Adenosine receptor(s) mediating negative inotropy of paced left atria, isoprenaline-stimulated paced left atria and papillary muscles, and negative chronotropy of spontaneously beating right atria were characterized. 2. Isometric tension of guinea-pig isolated paced left atria and left ventricular papillary muscles and rate of contraction of spontaneously beating right atria were recorded. Papillary muscles were pre-stimulated with isoprenaline (1x10-8 M). Concentration-response curves (CRCs) for tension or rate reduction by N6 cyclopentyladenosine (CPA), the stereoisomers of N6-(2-phenylisopropyl)adenosine ((+)-PIA and (-)-PIA), 5'-(N-carboxamido)adenosine (NECA), N6-2-(4 aminophenyl)ethyladenosine (APNEA) and N6-(3-iodobenzyl)adenosine-5'-N methyuromide (IB-MECA) revealed a potency order of CPA=(-)-PIA>NECA in right atria and papillary muscles, which is consistent with involvement of A1 receptors. The potency order in left atria was CPA=NECA>(-)-PIA>(+)-PIA>APNEA, which is not typical of A1 adenosine receptors. Weak activity of APNEA and IB MECA discounts involvement of A3 receptors. 3. pA2 values for the antagonism of CPA by 8(p-sulfophenyl)theophylline (8-SPT) were calculated from Schild plots (log concentration-ratio against log 8-SPT concentration), the unity slopes of which indicated competitive antagonism. The pA2 value in the papillary muscles was significantly higher than for atrial preparations, indicating a possible difference in receptor characteristics between atrial and papillary muscle responses. 4. In left and right atria there was a limit to the displacement of the CPA CRCs at higher concentrations of 8-SPT. The 8-SPT-resistant component of the response is suggested to arise from duality of coupling of a common A1 receptor through either different G proteins or G protein subunits to independent transduction pathways. The results with papillary muscles can be explained by a typical A1 receptor coupled to a single transduction pathway. PMID- 10455320 TI - Glucocorticoids potently block tumour necrosis factor-alpha- and lipopolysaccharide-induced apoptotic cell death in bovine glomerular endothelial cells upstream of caspase 3 activation. AB - 1. Endothelial cell damage in glomeruli and kidney arterioles appears to play a pivotal role in glomerular inflammatory diseases. Glomerular endothelial cells, a specialized microvascular cell type involved in the regulation of glomerular ultrafiltration, die by apoptosis in response to tumour necrosis factor-alpha (TNF-alpha), TNF-alpha/basic fibroblast growth factor (bFGF), TNF alpha/cycloheximide, and bacterial lipopolysaccharide (LPS). Apoptotic cell death is characterized by extensive DNA cleavage, DNA ladder formation, and characteristic morphological alterations. 2. In search for apoptosis-preventing signals, we identified glucocorticoids as potent death preventing factors. Co treatment of cells with 10 nM dexamethasone and TNF-alpha, TNF-alpha/bFGF, TNF alpha/cycloheximide, or LPS blocked roughly 90% of apoptotic cell death in glomerular endothelial cells. 3. Similarly to dexamethasone (TNF-alpha- and LPS induced apoptosis are prevented with IC50 values of 0.8 and 0.9 nM, respectively), other synthetic and natural forms of glucocorticoids, such as fluocinolone, prednisolone, hydrocortisone, and corticosterone potently inhibited cell death with IC50 values of 0.2, 6, 50 and 1000 nM, for TNF-alpha and 0.7, 8, 100 and 500 nM for LPS, respectively. 4. Apart from glucocorticoids, mineralocorticoids such as aldosterone also blocked TNF-alpha/LPS-induced apoptosis (IC50 approximately 500 nM for TNF-alpha and approximately 500 nM for LPS), whereas sex hormones, i. e. beta-estradiol and testosterone remained without effect. 5. The protective effect of glucocorticoids (and mineralocorticoids) required glucocorticoid receptor binding as it could be antagonized by the glucocorticoid receptor antagonist RU-486. Concerning TNF alpha and LPS signal transduction, we found that dexamethasone efficiently prevented TNF-alpha- and LPS-induced activation of caspase-3-like proteases. Therefore, we postulate inhibitory mechanisms upstream of terminal death pathways. PMID- 10455319 TI - Calcium channels involved in the inhibition of acetylcholine release by presynaptic muscarinic receptors in rat striatum. AB - 1. The mechanism of the inhibitory action of presynaptic muscarinic receptors on the release of acetylcholine from striatal cholinergic neurons is not known. We investigated how the electrically stimulated release of [3H]-acetylcholine from superfused rat striatal slices and its inhibition by carbachol are affected by specific inhibitors of voltage-operated calcium channels of the L-type (nifedipine), N-type (omega-conotoxin GVIA) and P/Q-type (omega-agatoxin IVA). 2. The evoked release of [3H]-acetylcholine was not diminished by nifedipine but was lowered by omega-conotoxin GVIA and by omega-agatoxin IVA, indicating that both the N- and the P/Q-type (but not the L-type) channels are involved in the release. The N-type channels were responsible for approximately two thirds of the release. The release was >97% blocked when both omega-toxins acted together. 3. The inhibition of [3H]-acetylcholine release by carbachol was not substantially affected by the blockade of the L- or P/Q-type channels. It was diminished but not eliminated by the blockade of the N-type channels. 4. In experiments on slices in which cholinesterases had been inhibited by paraoxon, inhibition of [3H]-acetylcholine release by endogenous acetylcholine accumulating in the tissue could be demonstrated by the enhancement of the release after the addition of atropine. The inhibition was higher in slices with functional N-type than with functional P/Q-type channels. 5. We conclude that both the N- and the P/Q-type calcium channels contribute to the stimulation-evoked release of acetylcholine in rat striatum, that the quantitative contribution of the N-type channels is higher, and that the inhibitory muscarinic receptors are more closely coupled with the N-type than with the P/Q-type calcium channels. PMID- 10455321 TI - Bronchodilator and anti-inflammatory activities of glaucine: In vitro studies in human airway smooth muscle and polymorphonuclear leukocytes. AB - 1. Selective phosphodiesterase 4 (PDE4) inhibitors are of potential interest in the treatment of asthma. We examined the effects of the alkaloid S-(+)-glaucine, a PDE4 inhibitor, on human isolated bronchus and granulocyte function. 2. Glaucine selectively inhibited PDE4 from human bronchus and polymorphonuclear leukocytes (PMN) in a non-competitive manner (Ki=3.4 microM). Glaucine displaced [3H]-rolipram from its high-affinity binding sites in rat brain cortex membranes (IC50 approximately 100 microM). 3. Glaucine inhibited the spontaneous and histamine-induced tone in human isolated bronchus (pD2 approximately 4.5). Glaucine (10 microM) did not potentiate the isoprenaline-induced relaxation but augmented cyclic AMP accumulation by isoprenaline. The glaucine-induced relaxation was resistant to H-89, a protein kinase A inhibitor. Glaucine depressed the contractile responses to Ca2+ (pD'2 approximately 3.62) and reduced the sustained rise of [Ca2+]i produced by histamine in cultured human airway smooth muscle cells (-log IC50 approximately 4.3). 4. Glaucine augmented cyclic AMP levels in human polymorphonuclear leukocytes challenged with N-formyl-Met-Leu Phe (FMLP) or isoprenaline, and inhibited FMLP-induced superoxide generation, elastase release, leukotriene B4 production, [Ca2+]i signal and platelet aggregation as well as opsonized zymosan-, phorbol myristate acetate-, and A23187 induced superoxide release. The inhibitory effect of glaucine on superoxide generation by FMLP was reduced by H-89. 5. In conclusion, Ca2+ channel antagonism by glaucine appears mainly responsible for the relaxant effect of glaucine in human isolated bronchus while PDE4 inhibition contributes to the inhibitory effects of glaucine in human granulocytes. The very low PDE4/binding site ratio found for glaucine makes this compound attractive for further structure-activity studies. PMID- 10455322 TI - Regulation of platelet function by catecholamines in the cerebral vasculature of the rabbit. AB - 1. 111In-labelled platelets were monitored continuously in the cerebral and pulmonary vascular beds of anaesthetized rabbits. Dopamine can, depending upon the concentration, either potentiate or inhibit thrombin-induced platelet accumulation in the cerebral vasculature of rabbits by unknown mechanisms. The effects of specific adrenergic and dopaminergic receptor antagonists were tested upon dopamine's actions on intracarotid (i.c.) thrombin-induced (80 u kg-1) platelet accumulation in the cerebral vasculature. The effect of adrenaline on the response to thrombin in this vascular bed was also investigated. 2. Thrombin induced platelet accumulation was significantly (P<0.01) potentiated by dopamine (100 microgkg-1 min-1, i.c.) and this effect was significantly inhibited by infusion of the alpha-adrenoceptor antagonist, phentolamine. 3 A higher dose of dopamine (2 mg kg-1 min-1, i.c.) inhibited thrombin-induced platelet accumulation. The beta-adrenoceptor antagonist, propranolol, did not significantly alter this inhibitory effect whereas it was abolished by the dopamine D1 selective antagonist, SCH23390. 4 Adrenaline (when administered i.c. by bolus injection or infusion) had no significant effect on thrombin-induced accumulation at any of the doses tested. 5 Potentiation of in vivo platelet accumulation by dopamine therefore seems to occur via alpha-adrenergic receptors. However, the inhibitory effect of dopamine appears to be exerted via the activation of dopamine D1 receptors and not via beta-adrenergic receptors. Our findings confirm that dopamine, but not adrenaline, can modify platelet function in the cerebral vasculature and these observations may have implications for current and potential therapeutic uses of dopamine and selective dopaminergic compounds. PMID- 10455323 TI - Spontaneously hypertensive rats cholinergic hyper-responsiveness: central and peripheral pharmacological mechanisms. AB - 1. The mechanisms and the subtypes of muscarinic receptors implicated in the cardiovascular effects of physostigmine were investigated in conscious normotensive and spontaneously hypertensive rats. 2. Intravenous (i.v.) physostigmine (50 microg kg-1) induced in both strains a long pressor response, accompanied by a bradycardia. This pressor response was larger in spontaneously hypertensive (+41+/-6 mmHg) than in Wistar-Kyoto (+25+/-2 mmHg) rats (P<0.05). 3. Pretreatment with atropine sulphate (0.4 mg kg-1 i.v.), completely abolished the physostigmine-induced pressor response in both normotensive and hypertensive rats. In both strains, the physostigmine pressor response was significantly reduced by the systemic administration of either an alpha1-adrenoceptor antagonist (prazosin, 1 mg kg-1) or a V1A-vasopressin receptor antagonist (AVPX, 20 microg kg-1). This physostigmine pressor effect was completely abolished in both strains when both antagonists were administered concomitantly. 4. In WKY rats, the pressor response to physostigmine (50 microg kg-1 i.v.) was inhibited in a dose-dependent manner by i. c.v. administration of atropine (ID50=3.70 nmoles), the M1 receptor antagonist pirenzepine (ID50=10.71 nmoles), the M2 receptor antagonist methoctramine (ID50=4.31 nmoles), the M3 receptor antagonist p-F-HHSiD (ID50=60.52 nmoles) and the M4 receptor antagonist tropicamide (ID50=214.20 nmoles). In the hypertensive strain, the ID50 were found to be significantly higher for atropine (7.34 nmoles), pirenzepine (21.60 nmoles) and p F-HHSiD (139.50 nmoles) (P<0.05). 5. The present results indicate that physostigmine acts in normotensive and spontaneously hypertensive rats, through stimulation of both central M2 and M1 cholinoceptors to induce a rise in blood pressure mediated by an increase in plasma vasopressin and sympathetic outflow. Moreover, our results suggest that some modifications of the M1 receptor subtypes in terms of expression or affinity could be responsible for the hyper responsiveness of the hypertensive strain to cholinomimetic agents. PMID- 10455324 TI - Effects of melatonin on rat pial arteriolar diameter in vivo. AB - 1. Based on our finding that melatonin decreased the lower limit of cerebral blood flow autoregulation in rat, we previously suggested that melatonin constricts cerebral arterioles. The goal of this study was to demonstrate this vasoconstrictor action and investigate the mechanisms involved. 2. The effects of cumulative doses of melatonin (10-10 to 10-6 M) were examined in cerebral arterioles (30 - 50 microM) of male Wistar rats using an open skull preparation. Cerebral arterioles were exposed to two doses of melatonin (3x10-9 and 3x10-8 M) in the absence and presence of the mt1 and/or MT2 receptor antagonist, luzindole (2x10-6 M) and the Ca2+-activated K+ (BKCa) channel blocker, tetraethylammonium (TEA+, 10(-4) M). The effect of L-nitro arginine methyl ester (L-NAME, 10-8 M) was examined on arterioles after TEA+ superfusion. Cerebral arterioles were also exposed to the BKCa activator, NS1619 (10(-5) M), and to sodium nitroprusside (SNP, 10-8 M) in the absence and presence of melatonin (3x10-8 M). 3. Melatonin induced a dose-dependent constriction with an EC50 of 3.0+/-0.1 nM and a maximal constriction of -15+/(-1%). Luzindole abolished melatonin-induced vasoconstriction. TEA+ induced significant vasoconstriction (-10+/(-2%). No additional vasoconstriction was observed when melatonin was added to the aCSF in presence of TEA+, whereas L-NAME still induced vasoconstriction (-10+/(-1%). NS1619 induced vasodilatation (+11+/(-1%) which was 50% less in presence of melatonin. Vasodilatation induced by SNP (+12+/(-2%) was not diminished by melatonin. 4. Melatonin directly constricts small diameter cerebral arterioles in rats. This vasoconstrictor effect is mediated by inhibition of BKCa channels following activation of mt1 and/or MT2 receptors. PMID- 10455325 TI - Effects of propranolol treatment on left ventricular function and intracellular calcium regulation in rats with postinfarction heart failure. AB - 1. Chronic treatment with beta-adrenergic blocking agents can improve survival in patients with heart failure. The mechanisms underlying the beneficial effects and whether these effects are generalizable to ischaemic heart failure are unresolved. 2. We performed echocardiographic-Doppler examinations in rats (n=28) 1 and 6 weeks after myocardial infarction (MI) or sham surgery. Rats were randomized to no treatment or propranolol (500 mg/l in drinking water) after the first echocardiogram. Isometric contractions and intracellular Ca transients were recorded simultaneously in noninfarcted left ventricular (LV) papillary muscles. 3. Untreated MI rats had significant LV dilatation (10.6+/-0.4* vs 8.9+/(-0.3) mm, MI vs control), impaired systolic function (fractional shortening=11+/-2* vs 38+/-2%), and a restrictive LV diastolic filling pattern. MI rats receiving propranolol had similar LV chamber sizes (10.6+/(-0.5) mm) and systolic function (13+/(-2%). The propranolol treated animals had higher LV end-diastolic pressures (27+/-2* vs 20+/(-3 mmHg) and a more restricted LV diastolic filling pattern (increased ratio of early to late filling velocities and more rapid E wave deceleration rate). Contractility of papillary muscles from untreated MI rats was depressed (1.6+/(-0.3) vs 2.4+/(0.5 g mm(-2). In addition, Ca transients were prolonged and the inotropic response to isoproterenol was blunted. Propranolol treatment did not improve force development (1.6+/(-0.3 g mm(-2) or the duration of Ca transients during isoproterenol stimulation. 4. Chronic propranolol treatment in rats with postinfarction heart failure did not improve LV remodeling or systolic function. LV diastolic pressures and filling patterns were worsened by propranolol. Treatment also did not produce appreciable improvement in contractility, intracellular Ca regulation or beta-adrenergic responsiveness in the noninfarcted myocardium. PMID- 10455326 TI - Regulation of epidermal homeostasis through P2Y2 receptors. AB - 1. Previous studies have indicated a role for extracellular ATP in the regulation of epidermal homeostasis. Here we have investigated the expression of P2Y2 receptors by human keratinocytes, the cells which comprise the epidermis. 2. Reverse transcriptase-polymerase chain reaction (RT - PCR) revealed expression of mRNA for the G-protein-coupled, P2Y2 receptor in primary cultured human keratinocytes. 3. In situ hybridization studies of skin sections revealed that P2Y2 receptor transcripts were expressed in the native tissue. These studies demonstrated a striking pattern of localization of P2Y2 receptor transcripts to the basal layer of the epidermis, the site of cell proliferation. 4. Increases in intracellular free Ca2+ concentration ([Ca2+]i) in keratinocytes stimulated with ATP or UTP demonstrated the presence of functional P2Y receptors. 5. In proliferation studies based on the incorporation of bromodeoxyuridine (BrdU), ATP, UTP and ATPgammaS were found to stimulate the proliferation of keratinocytes. 6. Using a real-time firefly luciferase and luciferin assay we have shown that under static conditions cultured human keratinocytes release ATP. 7. These findings indicate that P2Y2 receptors play a major role in epidermal homeostasis, and may provide novel targets for therapy of proliferative disorders of the epidermis, including psoriasis. PMID- 10455327 TI - KATP channels and 'border zone' arrhythmias: role of the repolarization dispersion between normal and ischaemic ventricular regions. AB - 1. In order to investigate the role of KATP channel activation and repolarization dispersion on the 'border zone' arrhythmias induced by ischaemia-reperfusion, the effects of glibenclamide and bimakalim, agents modifying action potential (AP) duration, were studied in an in vitro model of myocardial 'border zone'. 2. The electrophysiological effects of 10 microM glibenclamide and 1 microM bimakalim (n=8 each), respectively KATP channel blocker and activator, were investigated on guinea-pig ventricular strips submitted partly to normal conditions (normal zone, NZ) and partly to simulated ischaemic then reperfused conditions (altered zone, AZ). 3. By preventing the ischaemia-induced AP shortening (P<0.0001), glibenclamide reduced the dispersion of AP duration 90% (APD90) between NZ and AZ (P<0.0001), and concomitantly inhibited the 'border zone' arrhythmias induced by an extrastimulus (ES), their absence being significantly related to the lessened APD90 dispersion (chi2=8.28, P<0.01). 4. Bimakalim, which also reduced the APD90 dispersion (P<0.005) due to differential AP shortening in normal and ischaemic tissues, decreased the incidence of myocardial conduction blocks (25% of preparations versus 83% in control, n=12, P<0.05) and favoured 'border zone' spontaneous arrhythmias (75% of preparations versus 25% in control, P<0.05). 5. During reperfusion, unlike bimakalim, glibenclamide inhibited the ES-induced arrhythmias and reduced the incidence of the spontaneous ones (12% of preparations versus 92% in control, P<0.05), this latter effect being significantly related (chi2=6.13, P<0.02) to the lessened ischaemia-induced AP shortening in the presence of glibenclamide (P<0.0001). 6. These results suggest that KATP blockade may protect the ischaemic-reperfused myocardium from 'border zone' arrhythmias concomitantly with a reduction of APD90 dispersion between normal and ischaemic regions. Conversely, KATP channel activation may modify the incidence of conduction blocks and exacerbate the ischaemia-induced 'border zone' arrhythmias. PMID- 10455328 TI - Comparison of the functional potencies of ropinirole and other dopamine receptor agonists at human D2(long), D3 and D4.4 receptors expressed in Chinese hamster ovary cells. AB - 1. The aim of the present study was to characterize functional responses to ropinirole, its major metabolites in man (SKF-104557 (4-[2-(propylamino)ethyl]-2 (3H) indolone), SKF-97930 (4-carboxy-2-(3H) indolone)) and other dopamine receptor agonists at human dopamine D2(long) (hD2), D3 (hD3) and D4.4 (hD4) receptors separately expressed in Chinese hamster ovary cells using microphysiometry. 2. All the receptor agonists tested (ropinirole, SKF-104557, SKF-97930, bromocriptine, lisuride, pergolide, pramipexole, talipexole, dopamine) increased extracellular acidification rate in Chinese hamster ovary clones expressing the human D2, D3 or D4 receptor. The pEC50s of ropinirole at hD2, hD3 and hD4 receptors were 7.4, 8.4 and 6.8, respectively. Ropinirole is therefore at least 10 fold selective for the human dopamine D3 receptor over the other D2 receptor family members. 3. At the hD2 and hD3 dopamine receptors all the compounds tested were full agonists as compared to quinpirole. Talipexole and the ropinirole metabolite, SKF-104557, were partial agonists at the hD4 receptor. 4. Bromocriptine and lisuride had a slow onset of agonist action which precluded determination of EC50s. 5. The rank order of agonist potencies was dissimilar to the rank order of radioligand binding affinities at each of the dopamine receptor subtypes. Functional selectivities of the dopamine receptor agonists, as measured in the microphysiometer, were less than radioligand binding selectivities. 6. The results show that ropinirole is a full agonist at human D2, D3 and D4 dopamine receptors. SKF-104557 the major human metabolite of ropinirole, had similar radioligand binding affinities to, but lower functional potencies than, the parent compound. PMID- 10455329 TI - Microtubule cytoskeleton involvement in muscarinic suppression of voltage-gated calcium channel current in guinea-pig ileal smooth muscle. AB - 1. Effects of agents, which affect microtubule polymerization-depolymerization cycle, on Ba2+ current (IBa) flowing through voltage-gated Ca2+ channels and carbachol (CCh)-induced sustained suppression of IBa were examined in whole-cell voltage-clamped smooth muscle cells of guinea-pig ileum. 2. offchicine (100 microM) and vinblastine (100 microM), microtubule depolymerizers, increased the ampitude of IBa. Lumicolchicine (100 microM), an inactive analogue of colchicine, had no effect on IBa. 3. Taxol (1 - 100 microM), a microtubule polymerizer, decreased IBa in a concentration-dependent manner and accelerated the rate of inactivation of IBa. Baccatin III (100 microM), an inactive analogue of taxol, had no effect on IBa. 4. Colchicine (100 microM) and vinblastine (100 microM), but not lumicolchicine (100 microM), decreased or abolished the sustained component of CCh (10 microM)-induced IBa suppression. 5. Pretreatment with taxol (10 - 100 microM) resulted in a concentration-dependent decrease in IBa and the action of CCh on IBa. The inhibitory effects of taxol and CCh on IBa were not additive. 6. Colchicine (100 microM) or taxol (100 microM) had no effect on voltage-gated K+ channel current or CCh-induced non-selective cationic channel current. 7. These results suggest that polymerization of microtubules leads to suppression of Ca2+ channel activity, and that muscarinic sustained suppression of Ca2+ channel current is mediated by a signal transduction element which involves microtubule cytoskeleton. PMID- 10455330 TI - Nociceptin-induced scratching, biting and licking in mice: involvement of spinal NK1 receptors. AB - 1. Intrathecal (i.t.) injection of nociceptin at small doses (fmol order) elicited a behavioural response consisting of scratching, biting and licking in conscious mice. Here we have examined the involvement of substance P-containing neurons by using i.t. injection of tachykinin neurokinin (NK)1 receptor antagonists and substance P (SP) antiserum. 2. Nociceptin-induced behavioural response was evoked significantly 5 - 10 min after i.t. injection and reached a maximum at 10 - 15 min. Dose-dependency of the induced response showed a bell shaped pattern from 0.375 - 30.0 fmol, and the maximum effect was observed at 3.0 fmol. 3. The behavioural response elicited by nociceptin (3.0 fmol) was dose dependently inhibited by intraperitoneal (i.p.) administration of morphine. 4. The NK1 receptor antagonists, CP-96,345, CP-99,994 and sendide, inhibited nociceptin-induced behavioural response in a dose-dependent manner. A significant antagonistic effect of [D-Phe7, D-His9]SP (6 - 11), a selective antagonist for SP receptors, was observed against nociceptin-induced response. The NK2 receptor antagonist, MEN-10376, had no effect on the response elicited by nociceptin. 5. Pretreatment with SP antiserum resulted in a significant reduction of the response to nociceptin. No significant reduction of nociceptin-induced response was detected in mice pretreated with NKA antiserum. 6. The N-methyl-D-aspartate (NMDA) receptor antagonists, dizocilpine (MK-801) and D(-)-2-amino-5 phosphonovaleric acid (APV) (D-APV), and L-NG-nitro arginine methyl ester (L NAME), a nitric oxide (NO) synthase inhibitor, failed to inhibit nociceptin induced behavioural response. 7. off present results suggest that SP-containing neurons in the mouse spinal cord may be involved in elicitation of scratching, biting and licking behaviour following i.t. injection of nociceptin. PMID- 10455331 TI - Effects of nociceptin and nocistatin on antidromic vasodilatation in hairless skin of the rat hindlimb in vivo. AB - 1. We tested whether nociceptin (NCE), the endogenous ligand of the opioid receptor-like 1 (ORL1) receptor, and nocistatin (NST), which reverses central NCE effects when applied intrathecally (i.t.), affect small-diameter afferent fibre mediated vasodilatation in rat hairless skin. 2. Female Wistar rats were vagotomized. Ongoing sympathetic vasoconstrictor activity was abolished by bilateral section of the lumbar sympathetic trunk between ganglia L2 and L3. Sensory axons were selectively stimulated in the dorsal root L5 by 20 electrical impulses supramaximal for activating C-fibres at 1 Hz. Blood flow was measured on the plantar skin of the left hind paw in the L5 dermatome using laser Doppler flowmetry. 3. NCE injected intravenously (i.v.) as single boluses (1, 10 and 100 nmol kg(-1) 7 - 8 min before dorsal root stimulation (n=6) dose-dependently decreased blood pressure and local vascular resistance and suppressed antidromic vasodilatation maximally by 47% (P<0.01). When NCE was injected 2 min before stimulation (n=3), antidromic vasodilatation was reduced by 64% after NCE (1 nmol kg-1) and totally, or almost totally, abolished after the two higher doses. 4. NST (1 - 100 nmol kg(-1) i.v., n=6) was without significant effect on blood pressure and cutaneous vascular resistance. Applied 5 (n=6) or 2 min (n=3) before stimulation it also did not affect antidromic vasodilatation. NST (100 nmol kg( 1) i.v.) applied shortly before an equimolar dose of NCE did not antagonize NCE effects on vascular resistance, blood pressure and antidromic vasodilatation (n=4). 5. In conclusion, NCE inhibits antidromic vasodilatation, a component of neurogenic inflammation, in rat skin while NST is without effect. NST, at the small-diameter sensory ending, is not an effective antagonist of NCE. PMID- 10455332 TI - Active intestinal elimination of ciprofloxacin in rats: modulation by different substrates. AB - 1. Two in vivo models, in the rat, were used to investigate, in the presence of different substrates, the overall and net intestinal elimination of ciprofloxacin: an open-intestinal perfusion model and an intestinal loop model respectively. 2. In the presence of quinidine, verapamil and cyclosporin (substrates of the P-glycoprotein (P-gp)), plasma AUCs of ciprofloxacin were 1.5 2 fold increased, while biliary clearance (1.5 - 2 fold), intestinal overall and net clearances (2 - 4 fold and 1.5 - 8 fold respectively) decreased. The weak effect obtained with cyclosporin as compared to verapamil and especially quinidine, suggests, for ciprofloxacin, the existence of transport systems distinct from the P-gp, as the OCT1 transporter which could be inhibited by quinidine. 3. With cephalexin and azlocillin, two beta-lactam antibiotics, plasma AUCs of ciprofloxacin increased and biliary and intestinal overall clearances decreased in a similar fashion (1.3 - 2 fold), suggesting the involvement of organic anion and/or cation transporters. 4. In the presence of structural analogues, the effect was dependent on the compound administered: Sparfloxacin had no effect on intestinal clearance of ciprofloxacin. In contrast, with pefloxacin, overall intestinal clearance of ciprofloxacin was decreased and net intestinal clearance increased. 5. The specificity of ciprofloxacin intestinal transport appears to be different from P-gp as outlined by the lack of competition with sparfloxacin, a P-gp substrate. Ciprofloxacin intestinal elimination seems to be mediated by organic anion and/or cation transporters and a mechanism sensitive to quinidine and verapamil. PMID- 10455333 TI - Control of vascular tone in isolated mesenteric arterial segments from hypertensive patients. AB - 1. Experimental hypertension is associated with several functional alterations of vascular endothelium and smooth muscle, but relatively few studies have examined the control of arterial tone in isolated vascular preparations from patients with essential hypertension. Therefore, we compared functional characteristics in vitro of distal ring segments of the mesenteric artery from 17 hypertensive and 22 normotensive humans. 2. Arterial constrictor responses induced by cumulative addition of Ca(2+) in the presence of noradrenaline (NA) were more effectively inhibited by the Ca(2+) entry blocker nifedipine (0.5 nM) in hypertensive than normotensive subjects (by 55.4+/-4.9, n=17 and 35.0+/(-5.2%), n=22, respectively). Also the contractions elicited by high concentrations of KCl were more effectively inhibited by nifedipine in arterial rings from hypertensive than normotensive patients (by 38.9+/(-3.7), n=17 and 20. 2+/(-4.6%), n=22, respectively). However, the concentration-response curves of contractions to NA, serotonin and KCl in the absence of nifedipine were similar between the study groups. 3. The concentration-response curves of endothelium-dependent relaxations to acetylcholine and Ca(2+) ionophore A23187, as well as of endothelium independent relaxations to the nitric oxide donor nitroprusside, beta adrenoceptor agonist isoprenaline and K+ channel opener cromakalim did not show any differences between the groups. Moreover, the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester (0.1 mM) almost abolished the relaxations to acetylcholine and Ca(2+) ionophore in both groups, indicating that these responses were largely mediated by nitric oxide. The function of arterial sodium pump was evaluated by relaxations elicited by the return of K+ upon contractions induced by K+-free solution. The rate of K+-relaxation was similar in hypertensive and normotensive arteries (for all these responses n=20 - 22 in the normotensive and 15 - 17 in the hypertensive group). 4. These results suggest abnormal function of voltage-dependent Ca(2+) channels in arterial smooth muscle of hypertensive patients, whereas vascular responses to endothelium-dependent and -independent vasodilators and classical contractile agents were similar between hypertensive and normotensive subjects. The present findings support the view that blockade of voltage-dependent Ca(2+) channels is an effective means of reducing arterial tone in essential hypertension. PMID- 10455334 TI - Pharmacological characterization of beta-adrenoceptors mediating relaxation of the rat urinary bladder in vitro. AB - 1. Isoproterenol relaxed KCl-precontracted rat bladder strips with a pD2 of 7.21 leaving a residual contractile response of 3.2% after 30 microM. The selective beta1-agonist, T-0509 (pD2 : 6.24, 10.1% residual contraction after 100 microM), beta2-agonist, terbutaline (pD2 : 5.43, 13.7% residual contraction after 100 microM), and beta3-agonists, BRL 37344A (pD2 : 6.60, 17.3% residual contraction after 100 microM), and SR 58611A (pD2 : 5.15, 34.0% residual contraction after 100 microM), also relaxed bladder strips. 2. The relaxant response to isoproterenol was weakly but significantly antagonized by 1 microM propranolol which produced a 3 fold shift of the concentration-response curve to the right, and significantly antagonized by the beta1-selective antagonist, metoprolol (10 microM, 3 fold shift), and the beta2-selective antagonist, butoxamine (100 microM, 6 fold shift). A combination of 10 microM metoprolol and 100 microM butoxamine caused a 15 fold shift of the concentration-response curve for isoproterenol to the right. Incubation with the beta3-antagonist, SR 59230A (1 microM), caused a 6 fold shift of the concentration response curve for isoproterenol to the right. 3. The non-conventional partial agonist, CGP 12177A, weakly relaxed KCl-precontracted bladder strips (pD2 : 3.31, 51.3% residual contraction after 300 microM); the relaxation was resistant to blockade by 1 or 10 microM propranolol. 4. In the presence of 200 microM propranolol, CGP 12177A (20 microM) or SR 59230A (10 microM) antagonized surmountably the relaxant effects of BRL 37344A. 5. The data suggest that rat urinary bladder body contains beta1, beta2, and beta3-adrenoceptors, all of which mediate relaxation. PMID- 10455339 TI - Cyclophosphamide metabolism in children with Fanconi's anaemia. AB - Although patients with Fanconi's anaemia (FA) exhibit a heightened sensitivity to DNA cross-linking agents, modified doses of CY continue to be used in their conditioning prior to BMT. We measured the pharmacokinetics and metabolism of CY in six children with FA using an established high performance thin layer chromatography technique. CY doses ranged between 5 and 20 mg/kg (median 10 mg/kg). The median CY clearance was 0.6 l/h/m2 (range 0.4-1.1 l/h/m2), t1/2 was 8.1 h (range 6.7-9.5 h) and volume of distribution was 0.19 l/kg (range 0.16-0.34 l/kg), respectively. These results contrast with those previously reported from a comparable group of non-FA children in whom the median CY clearance was 3.2 l/h/m2 (range 2-5 l/h/m2) (P = 0.035), t1/2 was 2.4 h (range 2-3.8 h) (P = 0.035) and volume of distribution 0.5 l/kg (range 0.26-0.95 l/kg) (NS). Unlike the control group in whom the presence of inactive metabolites of CY was common, metabolites could not be found in any FA patient. The enhanced sensitivity of children with FA to CY may in part result from altered drug metabolism. PMID- 10455340 TI - A novel minor histocompatibility antigen recognized by HLA-A31 restricted cytotoxic T lymphocytes generated from HLA-identical bone marrow donor lymphocytes. AB - Bulk cytotoxic T lymphocytes (CTL) were generated by in vitro stimulation of BMT donor lymphocytes with Philadelphia chromosome (Ph)-positive leukemic cells from an HLA-identical sibling patient. CTL were cytotoxic against the patient's leukemic cells as well as the EBV-lymphoblastoid cell line (EBV-LCL) generated from the patient's cells, suggesting that they recognize a minor histocompatibility antigen (mHAg). Subsequently, several CTL lines were established by a limiting dilution method and analyzed. One of these CTL lines, 16C12 CTL which used a single TCRbetaV3S1 for CD8 cells, lysed HLA-A31-positive leukemic cells and EBV-LCL, but not fibroblasts. The cytotoxicity against the patient's leukemic cells and EBV-LCL was blocked by anti-HLA-A31 moAb, anti-HLA class I moAb, and anti-CD8 moAb, suggesting that this mHAg was presented with HLA A31. The antigen recognized by 16C12 CTL seemed to be a novel mHAg, since HLA-A31 restricted antigen has not been reported to date and 16C12 CTL showed no cytotoxicity against EBV-LCL which probably express known mHAgs. CTL detecting this mHAg may play an important role in the GVL effect in HLA-A31-positive BMT patients. PMID- 10455341 TI - Cord blood banking in London: the first 1000 collections. AB - The London Cord Blood Bank was established with the aim of collecting, processing and storing 10000 unrelated stem cell donations for the significant number of children in the UK requiring transplantation, for whom a matched unrelated bone marrow donor cannot be found. Collection is performed at two hospitals by dedicated cord blood bank staff after delivery of the placenta. Mothers are interviewed regarding medical, ethnic and behavioural history by nurse counsellors and sign a detailed consent form. Donations are returned to the bank for processing. Volume reduction is undertaken by a simple, closed, semi automated blood processing system, with excellent recovery of progenitor cells. Units are cryopreserved and stored in the vapour phase of liquid nitrogen. Blood samples from mothers and cord blood donations are tested for the UK mandatory red cell and microbiology markers for blood donors. Donations are typed for HLA-A, B and DR at medium resolution (antigen split) level using sequence-specific oligonucleotide probing and sequence-specific priming techniques. The selection of collection hospitals on the basis of ethnic mix has proven effective, with 41.5% of donations derived from non-European caucasoid donors. Bacterial contamination of collections has been dramatically reduced by implementation of improved umbilical cord decontamination protocols. PMID- 10455342 TI - Delaying treatment with granulocyte colony-stimulating factor after allogeneic bone marrow transplantation for hematological malignancies: a prospective randomized trial. AB - The use of granulocyte colony-stimulating factor (G-CSF) has been established to improve hematological recovery after allogeneic bone marrow transplantation (BMT). The optimal timing to start with G-CSF has not been determined. This study investigates whether delayed use of G-CSF starting on day 6 is as safe and efficient as starting treatment with G-CSF immediately after BMT. Thirty-eight patients undergoing allogeneic BMT were randomized to either receive post transplant G-CSF treatment starting at day 1 or at day 6. The time to hematological recovery was monitored and the groups were compared with respect to peritransplant morbidity and mortality. Recovery of the neutrophil granulocyte counts (PMN) to >100/microl, >500/microl and >1000/microl was comparable in both groups. The nadir of the PMN counts after stopping G-CSF was also similar. There was no difference in the recovery of red blood cells and platelet counts and no difference between the two groups with respect to the number of febrile episodes, number of days with antibiotics or number of documented bacterial, fungal or viral infections. Delayed treatment with G-CSF resulted in a reduction of G-CSF treatment from 19 days to 14 days (P = 0.0017). Reducing the length of treatment by 5 days lowered G-CSF treatment costs by 26.3%. Therefore, postponing treatment with G-CSF has no influence on the hematological recovery after allogeneic BMT. There is an economical benefit of postponing G-CSF use without any clinical disadvantages. PMID- 10455343 TI - Autografting of highly purified peripheral blood progenitor cells following myeloablative therapy in patients with lymphoma: a prospective study of the long term effects on tumor eradication, reconstitution of hematopoiesis and immune recovery. AB - In a prospective study, we have investigated CD34+ selection of peripheral blood progenitor cells (PBPC) for autotransplantation in patients with lymphoma. Twenty six consecutive patients (10 follicular lymphomas, seven mantle cell lymphomas, seven B-CLL, two immunocytomas) were mobilized using chemotherapy plus G-CSF. Sufficient numbers of PBPC could be collected from 24 patients and were immunoselected with the semiautomated Isolex 300 (n = 17) or the fully integrated Isolex 300i (n = 7) devices. The selection products were assayed by PCR amplification of clonal CDRIII or t(14;18) rearrangements for residual tumor cell content. Residual disease and long-term hematopoietic and immune recovery were studied by assessing the following parameters at 3, 6, and 12 months post transplant: CDRIII or t(14;18) PCR, platelet count, lymphocyte subsets, serum IgG, serum IgA, and measles titer. With the Isolex 300 device 26% (10-65) of input CD34+ cells were recovered with a median purity of 89.2% (49.4-98.9) after CD34+ selection. The Isolex 300i device allowed significantly better recoveries (46% (22-86)) and purities of CD34+ cells (98.8% (92.2-99.2)). The overall purging efficacy was 3.2 (0.6-5.1) log. Twenty patients have been reinfused with CD34+ selected grafts after myeloablative preparation. Rapid engraftment occurred in all patients. With a median follow-up of 28 (19-42) months, 14 patients are alive without clinical or molecular evidence of disease recurrence, whereas five have relapsed and one additional patient shows persistent presence of the disease specific molecular marker without clinical progression. Cellular and serological parameters of hematopoietic and immune functions were largely normal at 12 months post-transplant including the measles titer which was present in all patients. Kinetics of immunohematopoietic recovery were similar to those of 12 control patients who had received unmanipulated PBPC during the same time period except for the recovery of CD4+ CD45RA+ T cells which was significantly delayed in the CD34+ group. During the first year post-transplant, transient monoclonal or oligoclonal gammopathies were observed in seven of 16 study patients. We conclude that CD34+ selection with the Isolex system allows preparation of highly purified CD34+ fractions and effective tumor cell depletion. The CD34+ products can be reinfused safely after myeloablative treatment and result in sustained hematopoietic and immune recovery. The fact that all patients retained their specific measles immunity suggests that myeloablative treatment with reinfusion of highly purified CD34+ PBPC is not immunoablative. PMID- 10455344 TI - Role of the intensive care unit in children undergoing bone marrow transplantation with life-threatening complications. AB - The role of support measures in the Intensive Care Unit for bone marrow transplant recipients has been controversial. Data from 176 pediatric bone marrow transplants were retrospectively analyzed to ascertain the probability, causes, risk factors and survival for life-threatening complications requiring intensive care. Ninety-two patients underwent allogeneic BMT and 84 autologous BMT between January 1991 and December 1995. Thirty-one ICU admissions were recorded. The most frequent causes were acute respiratory failure (n = 15, mostly interstitial pneumopathies), septic shock (n = 5) neurological disorders (n = 5) and heart failure (n = 2). The cumulative incidence of an ICU admission at 20 months post transplant in patients with an allogeneic BMT was 25.7% (CI: 16.4-35.1), compared with 10.8% (CI: 4.2-17.5) in those with an autologous graft (P = 0.04). ICU admission frequency was maximum during the first 2 months post-transplant. All complications in patients with autologous transplants appeared during the first 5 months post-transplant. Among patients with allogeneic grafts, four were later admitted to the ICU, at 7, 9, 12 and 20 months post-transplant, respectively. The main risk factor for ICU admission was acute GVHD grades III-IV. No differences were found between patients with allogeneic transplants with GVHD grades 0-II and those undergoing autologous transplant. In contrast, differences were highly significant between patients undergoing allogeneic transplants with GVHD grades III-IV and those with GVHD grades 0-II or autologous transplants. No differences were observed between allogeneic and autologous transplants in terms of causes for ICU admission, duration of stay, hours on mechanical ventilation, hours on inotropic drug therapy and numbers of organs failing. Neither were differences found in ICU discharge survival between patients with allogeneic (50%, CI: 29.1 70.9) and autologous (66.7%, CI: 29.9-89.1) transplants. ICU discharge survival in patients admitted for lung disease was 28.6% (CI: 12.1-65.6) but 76.5% (CI: 41.9-87.8) in patients admitted for other causes (P = 0.007). ICU discharge survival in mechanically ventilated patients was 46.2% (CI: 27.0-65.4), significantly lower than nonventilated patients (100%). Three-year survival in all transplanted patients admitted to the ICU was 29.7% (CI: 13.1-45.0) compared with 70.2% (CI: 62.7-77.6) in patients not requiring ICU admission (P<0.001). Although a complication requiring admission to the ICU is, as confirmed by multivariate analysis, an unfavorable factor in long-term survival of transplanted patients, it must be emphasized that three of every 10 patients admitted to the ICU were alive and well at 3 years. Intensive care support in these patients can be life-saving. PMID- 10455345 TI - Arterial blood oxygenation, maximum exercise capacity and oxygen transport in patients before and after autologous peripheral blood progenitor cell transplantation. AB - Several studies have reported a reduced pulmonary diffusion capacity for carbon monoxide after total body irradiation (TBI) and/or high-dose chemotherapy (HDCT) and autologous peripheral blood progenitor transplantation (APBPC). The present study describes the oxygenation of arterial plasma (paO2), the maximum exercise capacity (Wmax) and maximum oxygen transport (VO2max) before and after such treatment. Pulmonary gas exchange and paO2 were measured in 15 patients at maximum dynamic bicycle exercise, performed before and at 8+/-0.9 (s.e.m.) months after TBI/HDCT (n = 12) and HDCT (n = 3) followed by APBPC. Wmax was 169+/-14 and 157+/-15 watts (P>0.05) before and after therapy, respectively. VO2max, 1.8+/-0.1 l/min before treatment, fell to 1.6+/-0.1 l/min (P<0.05) after therapy. The maximum exercise paO2 was 13.2+/-0.4 kPa before and 13.6+/-0.4 kPa (P>0.05) after the treatment period. The findings indicate no significant reduction of Wmax or pulmonary diffusion capacity for O2 after TBI/HDCT and APBPC while VO2max fell modestly by approximately 11%. The study was restricted to patients who remained in remission. PMID- 10455346 TI - Spleen sizing by ultrasound scan and risk of pneumococcal infection in patients with chronic GVHD: preliminary observations. AB - Encapsulated bacteria infections (EBI) can cause severe complications after BMT, usually occurring in patients with chronic GVHD (cGVHD) and attributed to functional hyposplenism. Using ultrasound (US) scan, we measured spleen size in 22 patients transplanted from HLA identical siblings, with or without cGVHD. No patient had received TBI, spleen irradiation or penicillin prophylaxis. Results were correlated with occurrence of EBI during a mean follow-up of 55 months (range 7-93). In the group without cGVHD, the difference between pre- and post BMT spleen longitudinal diameters was not significant, and no patient developed EBI. In the cGVHD group, post-BMT spleen longitudinal diameters were significantly smaller than those pre-BMT (9.1+/-1.6 vs. 12.3+/-2.2; P = 0.0005). Out of four patients with cGVHD who showed a major spleen size reduction, two developed a severe infection (an overwhelming sepsis and a pneumococcal meningitis). In our small series, we found a borderline relationship between spleen size reduction and duration of cGVHD (P = 0.06), as well as an increased risk of life-threatening infection in patients with extensive cGVHD and hyposplenism as detected by US scan. We conclude that US scan may be useful to detect spleen size reduction following allogeneic BMT and that penicillin prophylaxis is to be strongly recommended in patients with extensive cGVHD and spleen size reduction, even in those who have not received total body or spleen irradiation. PMID- 10455347 TI - Natural killer (NK) cells prevent virus production in cell culture. AB - Natural killer (NK) cells (CD3-/CD16+/CD56+ lymphocytes) play an important role in early immune defense against viral infection, a fact which is of prime significance for heavily immunosuppressed patients after bone marrow transplantation. In this study we demonstrate that NK cells preferentially lyse human colon adenocarcinoma (Colo-205) tumor cells infected with herpes simplex virus type 1 (HSV-1) and vaccinia virus (VV) and autologous T cells infected with VV. This phenomenon was assessed by the viral infectious center (IC) method and compared with the results obtained by means of the standard 51Cr-release assay. Using the IC assay, we found that NK cells lyse virus infected cells at an early stage of infection, thereby preventing viral dissemination to neighboring cells. 51Cr-release assay verified by propidium iodide (PI) penetration showed that the early effects of NK mediated anti-viral activity are not the result of membrane damage. The effect of NK cells on HSV-1 infected Colo-205 cells appears to be independent of the level of expression of major histocompatibility complex (MHC) class I molecules while the killing of autologous VV-infected T cells correlates with a reduction in MHC class I expression. Our results suggest that additional factors besides MHC play a role in the regulation of NK cell-mediated lysis of virus infected cells. This may be of clinical importance in patients who are heavily immunosuppressed after bone marrow transplantation. PMID- 10455348 TI - A quality of life study in 20 adult long-term survivors of unrelated donor bone marrow transplantation. AB - There are few specific data available concerning quality of life (QOL) of survivors of unrelated donor bone marrow transplantation (UD-BMT). The procedure is expensive, difficult and is being employed increasingly yet we have little information concerning the QOL of survivors to justify this intervention. In this study, 20 long-term (>1 year post-BMT) survivors were studied with four self report questionnaires designed to assess quality of life, satisfaction with life, social support and employment status. Overall, satisfaction with life measures was above average but there was dissatisfaction with physical strength and appearance. The post-transplant employment data indicates that 60% of long-term survivors returned to full-time work and 15% to part-time work. Failure to return to work was not correlated with graft-versus-host disease (GVHD), relapse, age at or time since transplant. In general, there was a good correlation between the clinician's and patient's view of their health but the clinician's assessment of the patients mental health and energy was higher than the patients reported. Further research is required in the area of QOL post-UD-BMT. This will enable transplant physicians to counsel patients better pre-BMT and to evaluate fully the results achieved by different centres performing the procedure. PMID- 10455349 TI - Tularemia--an unusual cause of a solitary pulmonary nodule in the post-transplant setting. AB - We report a case of tularemia presenting as a solitary pulmonary nodule following syngeneic PBSC transplant. Seven months after undergoing a syngeneic PBSC transplant for AML, our patient presented with fever without localizing signs. Chest X-ray revealed a solitary pulmonary nodule. Culture of a CT guided needle aspiration revealed Francisella tularensis. The patient was successfully treated with ciprofloxacin. His fever resolved and clearance of the nodule was documented on a CT scan 2 months after diagnosis and initiation of treatment. To our knowledge, this is the only reported case of tularemia occurring in the post transplant setting. The possible relationship between transplant-induced immune dysfunction and the occurrence of this rare infection is discussed. PMID- 10455350 TI - Delayed engraftment and mixed chimerism after HLA-identical sibling donor BMT in Fanconi anaemia. AB - A 12-year-old girl with Fanconi anaemia (FA) received a bone marrow transplant from her HLA-identical brother following conditioning with cyclophosphamide (20 mg/kg), thoraco-abdominal radiation (TAI) (4 Gy) and equine anti-thymocyte globulin (ATG) (90 mg/kg). Engraftment was delayed and initially tenuous, and was followed by mixed chimerism (MC) over a follow-up period of 2 years. DNA analysis of engraftment was performed on whole peripheral blood and on separated granulocytes, B and T lymphocytes using PCR detection of CA tandem repeat polymorphisms. At 10 weeks post BMT, granulocytes were predominantly donor, but B and T lymphocytes recipient, in origin. Over the subsequent 90 weeks, granulocytes and B lymphocytes were donor-derived, whilst T cells showed persistent MC but with an increasing donor component. Marrow haemopoietic function (Hb, ANC and platelet count) improved gradually in parallel with a rise in the proportion of donor lymphocyte engraftment. We postulate that a population of recipient lymphocytes survived conditioning and in turn delayed the development of full donor chimerism. Although transient MC has been described after allogeneic BMT in FA, its association with delayed engraftment, and persistence for more than 1 year post BMT, has not been documented clearly. PMID- 10455351 TI - HTLV-I-associated myelopathy following allogeneic bone marrow transplantation. AB - A 39-year-old polytransfused patient with aplastic anemia acquired transfusion associated HTLV-I infection shortly before transplantation. The patient underwent allogeneic bone marrow transplantation and developed HTLV-I associated myelopathy 3 years later. Clinical abnormalities and a host of atypical findings are presented in the context of previous reports describing uncommon features of the disease. PMID- 10455352 TI - Successful treatment of resistant thrombotic thrombocytopenic purpura/hemolytic uremic syndrome with autologous peripheral blood stem and progenitor (CD34+) cell transplantation. AB - The first-line treatment of thrombotic thrombocytopenic purpura-hemolytic uremic syndrome (TTP-HUS syndrome) induces a response and survival rate of approximately 85%, even if a considerable number of patients relapse; nevertheless, a number of these patients are resistant to conventional management. Immunoablation followed by stem cell transplantation has been shown to be capable of inducing remissions in a large spectrum of experimental autoimmune disorders. We report here the case of a 20-year-old male patient with the TTP-HUS syndrome who was resistant to conventional treatment and was transplanted with autologous immunoselected CD34+ PBPC after conditioning with cyclosphosphamide, anti-T lymphocyte globulin and prednisone. Seven months after transplant the patient is alive and well, without any further treatment being given. PMID- 10455353 TI - Immunological reconstitution after cord blood transplantation for an adult patient. AB - We transplanted 4.1x10(7) unrelated umbilical cord blood cells into a 27-year-old patient suffering from transformed acute myelocytic leukemia. The thawing method was the same as described by Rubinstein et al (Proc. Natl. Acad. Sci. USA 1995; 92: 10119-10122). ANC reached over 500x10(9)/l on day 19, and the patient was free from RBC and platelet transfusion on days 26 and 38, respectively. Cytogenetic and molecular analysis after transplant revealed complete chimerism. The CD3+CD4+ lymphocyte count became greater than 100x10(9)/l at 5 weeks after transplantation. The CD3+CD8+ count became greater than 500x10(9)/l at 7 weeks and thereafter progressively increased in spite of administration of CYA. This immunological reconstitution pattern after umbilical cord blood transplantation was different from that after bone marrow transplantation, and resistance of CD3+CD8+ lymphocytes to CYA was the distinguishing characteristic. The rapid hematological recovery and immunological reconstitution may be attributed to the high dose of transfused nucleated cells and umbilical cord blood transplantation may become a promising method for treatment for such cases. PMID- 10455354 TI - Autologous CD34+ cells transplantation after FAMP treatment in a patient with CLL and persisting AIHA: complete remission of lymphoma with control of autoimmune complications. AB - A 48-year-old male with CLL and concomitant AIHA unresponsive to chlorambucil was treated with fludarabine. The remission of CLL and improvement of the AIHA was achieved, but the patient remained steroid dependent. Therefore, high-dose chemotherapy followed by CD34-selected autologous peripheral blood stem cells transplantation was performed and this led to long-term clinical, immunophenotypic and molecular remission with disappearance of AIHA. Twenty-three months later, the CLL recurred with signs of AIHA. In this patient with AIHA, HDC and selected CD34+ cells completely, though temporarily, controlled both CLL and associated immune complications. This case illustrates the potential application of this approach in the management of CLL patients with immune complications. PMID- 10455355 TI - Molecular remission in adult T cell leukemia after autologous CD34+ peripheral blood stem cell transplantation. AB - This report describes a patient with acute-type adult T cell leukemia/lymphoma (ATLL) successfully treated by autologous CD34+ peripheral blood stem cell transplantation after fractionated total body irradiation and high-dose cytarabine and cyclophosphamide. A newly established inverse polymerase chain reaction method was used to demonstrate the disappearance of ATLL clonal cells. The patient achieved a sustained molecular remission after transplantation, but died from opportunistic infection 4 months after transplantation. Thus, autologous CD34+ peripheral blood stem cell transplantation is promising for this type of malignancy. However, a prudent clinical attitude toward immunological fragility after transplantation is needed for better outcome. PMID- 10455356 TI - Hepatic veno-occlusive disease after stem cell transplantation in Japan. PMID- 10455357 TI - T cell-depleted allogeneic bone marrow transplantation as post remission therapy for acute myelogenous leukaemia. PMID- 10455358 TI - Transplantation of allogeneic CD34+ blood cells in leukemia or lymphoma patients at high risk of GVHD. PMID- 10455359 TI - Detection of contaminating tumor cells by RT-PCR in breast cancer patients. PMID- 10455360 TI - Immunological and molecular detection of tumor contamination in leukapheresis products from children with disseminated neuroblastoma: implication for evaluation of tumor cell depletion after CD34 selection. PMID- 10455361 TI - Ex vivo expansion of enriched CD34+ cells from neonatal blood in the presence of thrombopoietin, a comparison with cord blood and bone marrow. AB - Neonatal blood (NB) contains substantial numbers of stem and progenitor cells which decline rapidly after birth. Using a combination of cord blood (CB) and NB, we performed a successful, sibling transplant for a thalassaemia patient, leading to the proposal that NB could be used as an adjunct to CB for transplantation. This study was aimed at addressing the feasibility of expanding NB and thus minimizing the volume needed from a NB collection. In the presence of early acting cytokines interleukin-1beta (IL-1beta), IL-3, IL-6, stem cell factor (SCF), flt-3 ligand with and without thrombopoietin (Tpo), we compared the expansion capacity of CD34+ enriched cells from CB, NB and bone marrow (BM). Flow cytometry and colony-forming unit (CFU) analyses show that Tpo significantly increased the expansion of CD34+ cells from CB and NB to early and committed progenitors. No significant difference was observed between the expansion of CB and NB at 7, 14 or 21 days of culture in terms of CFU, CD34+ and CD61+ cell subsets. The expansion capacity of BM was significantly lower than that of NB or CB, possibly related to the low proportion of CD34+ CD38- cells observed at day 0. There was a relatively rapid expansion of NB which was evident at day 7 whilst the expansion of CB and BM remained low, suggesting a speedy maturation process in the postnatal infant. The expanded cells, being heterogeneous in their morphology and cell surface marker expression, were mostly of the myeloid lineage (CD45+, CD33+ and HLA-DR+). Our results showed that the expansion capacity of NB is comparable to that of CB and if transplanted, the expanded products of NB might contribute to the engraftment kinetics of the neutrophil and megakaryocyte lineage. PMID- 10455362 TI - Bone marrow transplantation for transfused patients with severe aplastic anemia using cyclophosphamide and total lymphoid irradiation as conditioning therapy: long-term follow-up from a single center. AB - In transfused patients with aplastic anemia, incidence of graft rejection remains significant. Seventeen transfused patients with severe aplastic anemia received BMT from HLA-identical sibling donors after conditioning with cyclophosphamide (CY, 50 mg/kg/day for 4 days) plus total lymphoid irradiation (TLI, 750 cGy in a single dose). For graft-versus-host disease (GVHD) prophylaxis one patient received methotrexate, five patients received CsA and 11 received CsA in association with methylprednisolone. All patients had sustained engraftment. The actuarial survival of patients was 76% with a median follow-up for surviving patients of 11 years (range 0.3-14.5 years). The incidence of grade II-III acute GVHD was 24%, and chronic GVHD 35%. Median Karnofsky score of surviving patients is 100 (range 90-100). Only one patient developed interstitial pneumonia. None of the patients has developed a malignancy after BMT. The role of limited field irradiation in development of malignant neoplasms after BMT for aplastic anemia is discussed. We conclude that a conditioning regimen using CY + TLI in sensitized aplastic anemia patients results in a high survival rate on long-term follow-up. PMID- 10455363 TI - Chronic myeloid leukemia in first chronic phase not responding to alpha interferon: outcome and prognostic factors after autologous transplantation. EBMT Working Party on Chronic Leukemias. AB - We report the data obtained from the European Bone Marrow Transplant Registry for patients with CML who received autologous transplantation (AT) in chronic phase (CP) because alpha-IFN was ineffective. Forty-one CML patients (median age: 40.5 years; median Sokal index: 0.78) were included in this study. Bone marrow (16 cases) or blood (25 cases) progenitor cells were collected at diagnosis in 19 patients, during stable chronic phase or while the patient had cytogenetic (Cy) or complete hematologic response (CHR) in the other 22, and were manipulated ex vivo in 10 cases. The conditioning regimen consisted of busulfan associated with other chemotherapeutic regimens in 36 cases. Two patients died from interstitial pneumonitis (one case) and hemorrhage (one case). From the date of AT, the estimated probability of survival for the 41 patients was 84 +/- 13% and 51 +/- 29% at 2 and 4 years, respectively. Considering the 39 evaluable patients, the actuarial probability of achieving CHR, major and complete CyR 2 years after AT was 92 +/- 9%, 46 +/- 17%, and 30 +/- 15%, respectively. The Sokal score at diagnosis and the achievement of hematologic response after transplant were of prognostic importance. We suggest that a significant proportion of CML patients not responding to alpha-IFN may benefit from AT. PMID- 10455364 TI - Autologous stem cell transplantation in chronic myeloid leukemia: a single center experience. AB - Between 1980 and 1996, we transplanted 72 patients with CML using blood stem cells collected at diagnosis before treatment and without any mobilization. The median age of patients at diagnosis was 47.5 years (range 20.5-59.5). The median numbers of nucleated cells and CFU-GM transplanted were 10 x 10(8)/kg and 97 x 10(4)/kg, respectively. The median duration to reach more than 0.5 x 10(9)/l neutrophils and 50 x 10(9)/l platelets was 12 (range 5-19) and 11 days (range 0 79), respectively. Twenty patients (group I) were transplanted in chronic phase either for resistance to IFN (14 patients) (group IA) or because the Sokal index was more than 1.2 (six patients) (group IB). All those patients had preparative regimen with busulfan (4 mg/kg/day x 4) and melphalan (140 mg/m2). They were treated with recombinant alpha-interferon (IFN) after transplant. The cumulative incidence of major cytogenetic response (MCR) at 12 months was 25 +/- 21% (95% CI), the 5-year survival was 75 +/- 42% (95% CI). These results (observed in patients with bad prognosis factors) are similar to those usually observed in CML patients treated by IFN, whatever the Sokal risk. Thus autologous transplantation is able to reproduce for poor prognosis patients the results observed in standard risk patients treated with IFN. This suggests that it could prolong survival. Fifty-two other patients (group II) were transplanted for CML in transformation (accelerated phase = 32; blast crisis = 20) after a preparative regimen containing either total body irradiation (TBI) or busulfan. The median survival was short (10.4 months) and only 21 patients survived more than 1 year. The survival was longer for patients transplanted in accelerated phase (vs blast crisis), those who were due to receive a double transplant (vs single) (34 patients), those who were treated with IFN after transplant (vs hydroxyurea) and for the patients who obtained a complete hematologic response. PMID- 10455365 TI - Treatment outcome and prognostic factors for relapse after high-dose chemotherapy and peripheral blood stem cell rescue for patients with poor risk high grade non Hodgkin's lymphoma. AB - The aim of the study was to determine treatment outcome and identify a particularly high risk group in a consecutive series of 66 patients with poor prognosis high grade lymphoma (NHL) treated with conventional induction chemotherapy followed by high-dose chemotherapy (HDCT) and peripheral blood stem cells (PBSC) rescue. Fifty-one patients with intermediate grade NHL (Kiel) and two or three adverse prognostic features as defined by the age-adjusted International Prognostic Index (IPI) received induction treatment with 7 weeks of doxorubicin, cyclophosphamide, vincristine, bleomycin, etoposide, prednisolone and methotrexate (VAPEC-B) followed by three cycles of ifosfamide/cytarabine. Fifteen patients with high grade Burkitt's and lymphoblastic NHL received 11 weeks of VAPEC-B followed by three cycles of high-dose methotrexate. HDCT for all 66 patients consisted of busulphan/cyclophosphamide followed by autologous PBSC rescue. Thirty-one patients (47%) received HDCT in first complete remission (CR/CRu) and 34 patients (52%) in first partial remission (PR) after conventional chemotherapy. Following HDCT, 42 patients (64%) were in CR/CRu, 19 patients (29%) in PR and one patient had progressive disease. There were four toxic deaths. After a median follow-up period of 27 months (range 7-73) in 46 surviving patients, the actuarial 3-year estimates of overall survival, event-free survival (EFS) and freedom from progression (FFP) were 67%, 65% and 70%, respectively. In univariate analysis, prognostic factors associated with reduced EFS were mediastinal bulk (P = 0.02), > or = 3 extra-nodal sites (P = 0.02), remission status prior to HDCT (P = 0.05), low albumin (P = 0.08) and raised ESR (P = 0.09). No significant difference was observed between patients with intermediate or high grade NHL or between patients with two or three adverse IPI features. Multivariate analysis identified mediastinal bulk (P = 0.01), > or = 3 extra nodal sites (P = 0.01) and low albumin (P = 0.03) as joint predictors of poor EFS. Remission status prior to HDCT was not found to be significantly associated with reduced EFS, FFP or survival, suggesting early introduction of HDCT may benefit patients with a PR. Based on these three adverse features, three groups (0, 1 or > or = 2 features) could be identified with differing EFS, survival and freedom from progression (FFP) rates at 3 years; 85%, 63% and 20%, respectively for EFS, 84%, 64% and 25% for survival and 85%, 66% and 33%, respectively for FFP. This prognostic model may identify patients with a particularly poor prognosis despite HDCT, who may benefit from other therapeutic approaches. PMID- 10455366 TI - Unrelated donor stem cell transplantation after autologous transplantation: experience of a single center. AB - Patients who do not respond to autologous stem cell transplantation (ASCT) have a poor prognosis. Concerns about toxicity limit the use of unrelated donor stem cell transplantation (UDSCT), but the knowledge about outcome after UDSCT post ASCT is limited. We carried out a retrospective analysis of the outcome in seven consecutive patients with leukemia (n = 5), myeloma (n = 1) and graft failure (n = 1) who underwent UDSCT after ASCT. Donors were matched for HLA-A, -B and -DR (n = 6) or had one class I antigen mismatch (n = 1). Tissue typing was performed by a high-resolution genomic technique for class II. Median patient age was 34 (11 54) years and time from ASCT to UDSCT was 16 (3-22) months. Patients with malignant diseases were given TBI and a CY preparatory regimen. In addition, all patients received T cell antibodies prior to UDSCT. Grade I acute GVHD developed in all seven patients, but there was no sign of more severe acute GVHD. Two of four evaluable patients developed limited chronic GVHD. Three died of transplant related toxicity, all due to pulmonary complications. Four patients are alive at 1.1, 1.5, 3.1 and 4.9 years post-UDSCT. A closely matched UDSCT could be considered for selected patients who are not cured by an ASCT. PMID- 10455367 TI - Comparison of the classic Glucksberg criteria and the IBMTR Severity Index for grading acute graft-versus-host disease following HLA-identical sibling stem cell transplantation. International Bone Marrow Transplant Registry. AB - Acute graft-versus-host disease (AGVHD) severity is usually graded (grades 0-IV) by the pattern of organ involvement using the classic Glucksberg-Seattle criteria (GSC). Recently, the International Bone Marrow Transplant Registry (IBMTR) developed a new Severity Index by regrouping the patterns of organ involvement into five Indexes (0-D) that appeared more predictive of transplant-related mortality (TRM) and transplant failure (TF, relapse or TRM). We studied the predictive value of both grading systems of TRM, TF and GVHD-related mortality (GTRM) in a series of 114 consecutive patients > or = 12 years old allografted from a histocompatible sibling at our institution, 100 of whom were evaluable for AGVHD. The IBMTR Severity Index showed better incremental prediction of TRM (relative risks (RR) of 1, 1.5, 1.4, 2 and 2.5 for Indexes 0, A, B, C and D), TF (RRs of 1, 1.6, 1.6, 2 and 2.3, respectively) and GTRM (RRs of 1, 2.2 and 4.8 for Indexes B, C and D) than the GSC. With the GSC different outcomes for TRM and TF were found only from grade 0 to I-II and 0 to IV or I-III to IV, but not from I II to III. The GSC also appeared less predictive of GTRM (RRs of 1, 0.4 and 2.9 for grades II, III and IV). In our relatively small patient sample, the new IBMTR Severity Index appeared more predictive of transplant outcome than the GSC, especially between no AGVHD, early Indexes (A-B) and advanced Indexes (C-D). PMID- 10455368 TI - Factors that influence long-term hematopoietic function following autologous stem cell transplantation. AB - The aim of the present study was to assess which factors influence hematopoietic function long term after transplantation. For this purpose, we have analyzed a series of 79 patients who underwent autologous transplantation. None of them received any further chemotherapy or radiotherapy after transplant. All patients were disease-free 1 year after autologous transplantation. Late impairment of hematopoietic function was defined as the presence of non-transient peripheral blood cytopenias, detected 6 and 12 months after autografting. Before transplantation, 38.7% of patients showed peripheral blood cytopenias. Six and 12 months after transplantation, cytopenias presented in 44.2% and 42.4% of patients, respectively. Cases displaying cytopenias 6 months after transplantation had received a significantly lower dose of CFU-GM and CD34+ cells than patients without cytopenias (P = 0.012 and P = 0.04, respectively). The same correlation, with even higher statistical significance, was observed 12 months after transplant (P = 0.007 and P = 0.005). Alkylating agents and radiotherapy administered prior to transplantation and age did not seem to influence the presence of permanent cytopenias. The incidence did not vary significantly according to the stem cell source (bone marrow or peripheral blood). The number of CFU-GM and CD34+ cells infused was the most important factor for maintenance of adequate hematopoiesis. PMID- 10455369 TI - Quantitative lymphocyte subset reconstitution after allogeneic hematopoietic transplantation from matched related donors with CD34+ selected PBPC grafts unselected PBPC grafts or BM grafts. AB - CD34+ cell selection of PBPC after harvest from G-CSF-treated allogeneic donors results in a more than 200-fold depletion of T lymphocytes in the graft and has been used to reduce the incidence of acute GVHD post transplant. Since transplantation with T cell-depleted BM grafts is associated with a delay in immune reconstitution and an increase of opportunistic infections, we evaluated the immunological reconstitution of patients with hematologic malignancies after therapy followed by CD34+-selected PBPC34 transplantation from matched related donors. Lymphocyte subset reconstitution over the first 12 months post transplant and the incidence of infections were evaluated in 12 patients receiving PBPC34 grafts and compared to that of patients after transplantation with PBPC without CD34+ enrichment (n = 20) or unmanipulated bone marrow grafts (BM; n = 15). PBPC34 grafts contained 264-fold fewer T lymphocytes (median 0.53 x 10(6) kg/body weight) than PBPC grafts and 36-fold fewer than BM grafts (140 x 10(6)/kg and 19 x 10(6)/kg, respectively). Despite a two log depletion of T cells in the PBPC34 grafts, T lymphocyte reconstitution appeared comparable among the three transplant groups over the first 12 months. A positive patient CMV serostatus pretransplant was correlated with a faster T cell reconstitution in all transplant groups. GVHD prophylaxis with methylprednisolone delayed B lymphocyte reconstitution. The incidence of infections post transplant did not appear to be increased in the PBPC34 group compared with the PBPC and BMT groups. It remains to be shown in larger prospective trials, whether these promising preliminary data of lymphocyte reconstitution and the clinical course after transplantation with PBPC34 can be confirmed. PMID- 10455370 TI - Thrombotic microangiopathy following allogeneic bone marrow transplantation. AB - Thrombotic microangiopathy is one of the complications of bone marrow transplantation and is related to other complications such as graft-versus-host disease, veno-occlusive disease, diffuse alveolar hemorrhage, and cytomegalovirus infection. Thrombotic microangiopathy occurred in three out of 12 patients who underwent allogeneic bone marrow transplantation over the past 1 year at our department. We compared the changes in cytokines and other molecules between patients with and without microangiopathy from before conditioning to the early post-transplantation period. All three patients with microangiopathy showed a significant increase of interleukin-12 at the time of leukocyte recovery after transplantation (two-way layout analysis of variance; P < 0.05), while none of the patients without microangiopathy showed an increase of interleukin-12. No significant differences were found between the two groups with respect to the other cytokines and molecules that were tested. These findings suggested that thrombotic microangiopathy might be predicted at an early stage after bone marrow transplantation by detecting an increase of interleukin-12 at the time of leukocyte recovery. The possibility that thrombotic microangiopathy is related to inflammation or autoimmunity was also suggested. PMID- 10455371 TI - Mucormycosis in allogeneic bone marrow transplant recipients: report of five cases and review of the role of iron overload in the pathogenesis. AB - In a 10-year consecutive series of 263 allogeneic bone marrow transplant recipients, we identified five cases (1.9%) of invasive mucormycosis. Only one infection occurred within the first 100 days after transplantation, while the remainder complicated the late post-transplant course (median day of diagnosis: 343). Sites of infection were considered 'non-classical' and included pulmonary, cutaneous and gastric involvement. No case of fungal dissemination was observed. Mucormycosis was the primary cause of death in three of the five patients. Corticosteroid-treated graft-versus-host disease, either acute or chronic, or severe neutropenia were present in all cases. However, compared with a matched control population, the most striking finding was the demonstration of severe iron overload in each of the mucormycosis patients. The mean level of serum ferritin, transferrin saturation and number of transfused units of red cells (2029 microg/l, 92% and 52 units, respectively) in the study group is significantly higher compared with the control group (P < 0.05). The difference with other risk groups for mucormycosis, including deferoxamine-treated dialysis patients and acidotic diabetics, was analyzed in view of the possible pathogenic role of iron. Although these infections are often fatal, limited disease may have a better prognosis if diagnosed early and treated aggressively. PMID- 10455372 TI - Factors associated with attrition from a national bone marrow registry. AB - During its 10-year existence, the National Marrow Donor Program (NMDP) has been extremely successful at recruiting potential bone marrow donors to join the volunteer registry. Due in part to successful recruitment and the longevity of the registry, the focus of the NMDP has now shifted to decreasing potential attrition when volunteers are recontacted for additional testing to determine whether they would be the optimal donor for a specific patient. Our own interest in the bone marrow donation process led us to examine four domains of variables - demographic characteristics, volunteer history, recruitment-related characteristics and donation-related concerns - that we hypothesized would be associated with increased likelihood of donor attrition at a key donor decision point (DR-stage blood typing). Questionnaires were mailed to potential donors after they were contacted at the DR-stage, and had made the decision of whether or not to continue with blood typing. Our final sample included 756 volunteers who decided to continue with typing, and 258 individuals who declined further participation in the registry. In the bivariate analyses, factors in three of the four domains (all except demographic characteristics) were found to be substantially correlated with likelihood of attrition. Logistic regression indicated that nine central variables across the three domains produced the majority of increased attrition likelihood. Finally, a dose-response analysis suggested that as the number of attrition-related factors endorsed by an individual increased, his/her likelihood of dropping out of the registry also increased. Implications for future research and interventions to reduce potential donor attrition are discussed. PMID- 10455373 TI - Immune thrombocytopenia after umbilical cord progenitor cell transplant: response to vincristine. AB - An 8-month-old male with X-linked lymphoproliferative disease underwent an unrelated, partially matched (with major mismatch at DR locus), cord blood stem cell transplant. Four months following the transplant, he developed immune thrombocytopenia with hemolytic anemia (Evans syndrome). He received multiple courses of intravenous immunoglobulin, anti-Rh D immunoglobulin, a pulse of high dose corticosteroids and cyclosporine with some improvement of hemolytic anemia, but no improvement of the thrombocytopenia. Addition of vincristine, resulted in long-term resolution of thrombocytopenia and anemia. No major toxicity was observed during treatment. Vincristine should be considered as a treatment for refractory immune thrombocytopenia after hematopoietic stem cell transplantation. PMID- 10455374 TI - Mycobacterial central venous catheter tunnel infection: a difficult problem. AB - We report our experience of non-tuberculous mycobacterial infection associated with the tunnel of Hickman-Broviac central venous catheters in immunosuppressed patients with haematological malignancies undergoing high-dose chemotherapy supported by BMT. The problem is rare and difficult to treat. Our cases are unique in developing tunnel site mycobacterial infection well after the tunnelled catheters were removed. We diagnosed one case of Mycobacterium chelonae, which is a well-documented cause of such infections, and two cases of Mycobacterium haemophilum, which are the first reported cases in this setting. Early wide surgical excision of the infected tunnel site and prolonged antibiotic therapy is necessary. Despite these measures recurrence occurred in two cases. Close liaison with the microbiology laboratory is needed to ensure the appropriate culture media and conditions are used for these fastidious organisms. Empiric antibiotic regimens should be based on the likely organism. Drugs active against M. chelonae and M. haemophilum should be included. PMID- 10455375 TI - Pneumatosis [correction of Pneumocystis] cystoides intestinalis with pneumoperitoneum and pneumoretroperitoneum in a patient with extensive chronic graft-versus-host disease. AB - Pneumatosis cystoides intestinalis is a rare finding of intramural gasfilled cysts in the bowel wall and sometimes free air in the abdomen. A few conditions are reported to cause this disease, one of them being immunosuppression. We describe a 50-year-old Caucasian male with extensive chronic graft-versus-host disease (GVHD) of the gut and skin who developed PCI with pneumoperitoneum and pneumoretroperitoneum. To our knowledge, this is the first report of PCI occurring in a patient with active chronic GVHD which resolved spontaneously. PMID- 10455376 TI - Autoimmune thrombocytopenia in a patient with small cell lung cancer developing after chemotherapy and resolving following autologous peripheral blood stem cell transplantation. AB - A 46-year-old white male with small cell lung cancer (SCLC) limited to the thorax developed autoimmune thrombocytopenic purpura (AITP), following a cyclophosphamide, paclitaxel and G-CSF-containing regimen for peripheral blood stem cell (PBSC) mobilization. AITP associated with small or non-small cell lung cancer has been reported. We considered that the AITP in this case may be a part of paraneoplastic syndrome, which is frequently seen in patients with SCLC. The patient received HDC and autologous PBSC transplantation (APBSCT) for SCLC and the AITP resolved following transplantation, thus supporting the concept of HDC + APBSCT for the treatment of autoimmune diseases. PMID- 10455377 TI - Donor lymphocyte infusion post-non-myeloablative allogeneic peripheral blood stem cell transplantation for chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is a primary immunodeficiency disease symptomized by failure to generate superoxide and recurrent bacterial and fungal infections. Allogeneic bone marrow transplantation (BMT) is one of the therapeutic options available. However, it presents considerable risk to the recipient, especially if the patient is already at an advanced stage of disease, after repeated bacterial and fungal infections and organ damage. We present a case report of a 6-year-old child with long-standing CGD, severe clubbing, and jeopardized pulmonary function after multiple bacterial pulmonary infectious episodes, who had failed treatment with sulphamethazole trimethoprim, multiple antibiotic courses, itraconazole, as well as steroid and interferon-y therapy. He underwent allogeneic peripheral blood stem cell transplantation (alloPBSCT) from his HLA-matched MLC non-reactive sister following non-myeloablative conditioning. His ANC did not fall below 0.2 x 10(9)/l, his lowest WBC was 0.6 x 10(9)/l, and his platelets did not fall below 28 x 10(9)/l. He had normal engraftment, with no mucositis or organ toxicity. Neither parenteral nutrition nor platelet infusions were necessary. Partial donor chimerism following alloPBSCT was converted to full donor chimerism and superoxide production reverted to normal after donor lymphocyte infusions (DLI) from his HLA-matched sister. Twenty four months post transplant the patient is well, with stable and durable engraftment, 100% donor chimerism, normal superoxide production, no GVHD, and stabilization of his pulmonary condition. We suggest that alloPBSCT preceded by non-myeloablative conditioning and followed by DLI may constitute a successful mode of therapy for patients suffering from advanced CGD with recurrent infectious episodes resulting in organ dysfunction, enabling them to achieve full donor chimerism and normal superoxide production with minimal risk of transplant-related toxicity and GVHD. PMID- 10455378 TI - Massive myelomatous ascites responsive to VAD chemotherapy and autologous stem cell transplantation. AB - Plasmacytic ascites is an infrequent complication of multiple myeloma. To date, only few cases have been reported with a very rapidly fatal course unresponsive to therapy. We describe a patient with plasmacytic ascites and quiescent multiple myeloma of 8 years of duration. Disease progression became apparent due to myelomatous ascites, unexplained fever, pancytopenia and bone marrow infiltration. This case showed a complete and long-lasting response after VAD chemotherapy followed by autologous PBSC transplantation. Ascites in association with MM may respond for lengthy periods to high-dose chemotherapy and ASCT. PMID- 10455379 TI - Arsenic and all-trans retinoic acid as induction therapy before autograft in a case of relapsed resistant secondary acute promyelocytic leukemia. AB - Arsenic trioxide has recently been reported to be successful in the treatment of promyelocytic leukemia. Several concerns about the use of this toxic agent are currently reducing its potential clinical use even in severely ill patients. In this report we describe the results achieved by As2O3 with all-trans retinoic acid in a patient suffering from secondary, relapsed, resistant promyelocytic leukemia. Several complications, including sepsis and an extensive area of skin necrosis, did not allow us to treat the patient further with chemotherapy. With As2O3 and ATRA therapy, the patient obtained a complete molecular remission without any significant toxicity and, subsequently, it was possible to perform a bone marrow autograft in a state of complete remission. PMID- 10455380 TI - Persistent infection with human T lymphotropic virus type 1 after allogeneic bone marrow transplantation from a seronegative donor. PMID- 10455381 TI - Fibrous dysplasia in the jaws of a Hong-Kong population: radiographic presentation and systematic review. AB - OBJECTIVE: To compare the radiographic presentation of fibrous dysplasia in the jaws of Hong Kong patients with the features reported in other populations. METHODS: The clinical records and radiographs were reviewed of seven patients with fibrous dysplasia whose diagnosis had been histopathologically confirmed. Published series of fibrous dysplasia were subjected to systematic review. RESULTS: The male:female ratio was 4:3: one woman was Indian in origin; the other six patients were ethnic Chinese. The females were on average older than the males. Three cases affected the mandible and four the maxilla. Five cases affected the right side and two the left. A swelling was the principle clinical manifestation. The lesions were generally large, affecting most or all of the hemimandible or hemimaxilla involved. All cases exhibited expansion and had ground glass opacification. The four cases affecting the maxilla reduced the lumen of the maxillary antrum. The systematic review was carried out on 104 individual cases derived from nine reports of which only 93 cases were accompanied by radiological details. Many of the reports were limited in their scope, particularly with regards to radiological features. Fibrous dysplasia is more frequent in the maxilla in Oriental populations. Swelling is the most frequent clinical finding and buccolinqual expansion the most frequent radiological finding. CONCLUSION: The pattern of presentation of fibrous dysplasia in a Hong Kong population is broadly in agreement with reports from other populations. PMID- 10455382 TI - Influence of the digital image display monitor on observer performance. AB - OBJECTIVE: To assess the influence of the display monitor on observer performance. MATERIAL AND METHODS: Artificial enamel lesions were created in 40 extracted teeth at random using 1/4 and 1/2 round burs. Teeth were mounted in dental stone blocks to simulate a hemidentition. Approximate exposures were recorded at 70 kVp using a Digota (Soredex, Orion Corp, Helsinki, Finland) digital imaging system, calibrated to achieve optimum density. Six dentists rated each image on a five-point scale for the presence or absence of a lesion. Radiographic images were viewed on the following monitors: (1) AlphaScan 711 (Sampo Corp.); (2) Multiscan 17 Se II (Sony Electronics Inc.); (3) DS 2000 (Clinton Electronics Corp.) and (4) Latitude CP Laptop (Dell Computer Corp.). Raters were allowed to magnify and to adjust density and contrast of each image at will. Receiver Operating Characteristic (ROC) analysis was performed and curves were plotted for each image. Data was subjected to repeated measures analysis of variance and ordinal logistic regression to test for significance between variables and to determine odds ratios. RESULTS: Mean ROC curve areas ranged from 0.8728 for the Sampo monitor to 0.8395 for the Sony. Repeated measures analysis of variance showed significant differences between observers (P < 0.0001), lesion size (P < 0.0001), examiner/monitor interaction (P < 0.033) and examiner/block interaction (P < 0.013). However, no significant difference was found between monitors. CONCLUSION: This study suggests that observer performance is independent of the visual characteristics of the display monitor. PMID- 10455383 TI - The effect of developer age on the detection of approximal caries using three dental films. AB - OBJECTIVES: To compare the diagnostic accuracy for the detection of approximal caries of three dental X-ray films using fresh and aged processing chemicals. METHODS: Fifty-six extracted unrestored premolars were radiographed under standardized conditions using the new Dentus M2 (Agfa-Gevaert, Mortsel, Belgium), Ektaspeed Plus and Ultra-speed (Kodak Eastman Co, Rochester, USA) dental films. The films were processed manually using Agfa chemicals (Heraeus Kulzer, Dormagen, Germany). The procedure was repeated once a week until the complete exhaustion of the chemicals (6 weeks). Three independent observers assessed 210 radiographs using the following rating scale: 0 = sound, 1 = enamel lesion; 2 = lesion reaching the ADJ; 3 = dentinal lesion. True caries depth was determined by histological examination (14 sound surfaces, 11 enamel lesions, eight lesions reaching the ADJ and 23 dentinal lesions). True caries depth was subtracted from the values given by the observers and an analysis of variance was performed. The null hypothesis was rejected when P < 0.05. RESULTS: No significant differences were found in the diagnostic accuracy between the three films when using chemicals of up to 3 weeks old (P = 0.056). After the third week, Ultra-speed was significantly better than the other two films (P = 0.012). On average caries depth was underestimated. CONCLUSION: A similar level of diagnostic accuracy for approximal caries is achieved when using the three films. Dentus M2 and Ektaspeed Plus are at present the fastest available films and they should therefore be recommended for clinical practice. Agfa chemicals should be renewed every 3 weeks. Fifty per cent reduction in average gradient is indicative of renewing processing chemicals. PMID- 10455384 TI - The effects of sustained incisal clenching on the temporomandibular joint space. AB - OBJECTIVE: To determine the effect of sustained incisal clenching on the width of the temporomandibular joint space. METHODS: Nine normal subjects clenched on an anterior appliance for 10 min at 49 N. Sagittal tomograms were obtained during comfortable closure in the intercuspal position (ICP) (ICP-Before), comfortable closure on bite force transducer without clenching (BFT-Before), start of clenching (BFT-0), end of 5 min clenching (BFT-5), end of 10 min clenching (BFT 10), comfortable closure on the bite force transducer immediately after clenching (BFT-After) and comfortable closure in ICP after clenching (ICP-After). Joint space dimensions were automatically measured by a computerized image analysis system. RESULTS: The minimum joint space dimension was significantly reduced at BFT-5 (P = 0.0381), BFT-10 (P = 0.0019) and BFT-After (P = 0.0053) in relation to BFT-Before. The condylar position was also significantly shifted upward at BFT-0 (P = 0.0422), BFT-5 (P = 0.0005), BFT-10 (P = 0.0001), and BFT-After (P = 0.0004) in relation to BFT-Before. CONCLUSION: Sustained incisal clenching at 49 N causes significant anterior joint space reduction. We believe this is due to marked compression of the articular cartilage and disc. PMID- 10455385 TI - Relationship between bone mineral density and tooth loss in elderly Japanese women. AB - OBJECTIVE: To evaluate the relationship between vertebral bone mass and tooth loss and jaw bone mass in elderly Japanese women. METHODS: Mandibular cortical bone mass, alveolar bone height and number of teeth present (total, anterior, and posterior) were compared with the 3rd lumbar vertebral bone mineral density (L3BMD), measured by dual energy computed tomography (DEQCT), in 90 Japanese women by means of multiple regression analysis, controlling for body mass index, menopausal status, years since menopause and self-reported periodontal condition. RESULTS: Mandibular cortical bone mass and number of posterior teeth were associated with both alveolar bone height and L3BMD, but there was no association between alveolar bone height, number of anterior teeth present and L3BMD. CONCLUSION: Our results suggest that the loss of posterior teeth may be associated with a decrease not only in alveolar bone height, but also alveolar bone mineral density (BMD). The latter may be related to a decrease of lumbar vertebral BMD. PMID- 10455386 TI - Prediction of perceptibility curves of direct digital intraoral radiographic systems. AB - OBJECTIVES: To derive and test a method to predict Perceptibility Curves (PCs) for direct digital intraoral radiographic systems. METHODS: A test object was used to determine the performance of an average observer with respect to the threshold contrast and internal noise of the human visual system. These data were combined with system parameters to predict PCs mathematically. RESULTS: Data on the performance of an average observer could be defined to obtain values of the effective threshold contrast as a function of background gray level. This function combined with the gamma-value of the system predicted PCs that agreed well with PCs obtained in the conventional way. CONCLUSION: It is possible to predict PCs from a limited number of system parameters together with predetermined data on an average observer. PMID- 10455387 TI - Cross-sectional presurgical implant imaging using tuned aperture computed tomography (TACT). AB - OBJECTIVES: To investigate tuned aperture computed tomography (TACT) as an alternative to conventional tomography for cross-sectional imaging of potential implant sites. METHODS: Twenty potential implant sites in three dry human mandibles were selected. Sixteen TACT basis projections were recorded on film using a linear array of source positions. Films were scanned and 20 TACT and iterative TACT slices were reconstructed using TACT Workbench software. Maximum height and width were measured on one representative slice of each site. Results were compared with the ground truth using ANOVA and Wilcoxon signed rank tests. RESULTS: The means of relative difference in maximum height were 1.55 and 1.56% for TACT and iterative TACT respectively (P = 0.8643). The means of relative difference in width were 5.96 and 6.36% for TACT and iterative TACT respectively (P = 0.4845). More than 75% measurements were within +/- 1 mm of ground truth for both modalities. CONCLUSIONS: TACT may provide an alternative to conventional tomography for dental implant imaging. Further investigations should be performed to explore factors affecting image quality. PMID- 10455388 TI - Progression of maxillofacial squamous cell carcinoma evaluated using computer graphics with spiral computed tomography. AB - This report describes the appearance of maxillofacial squamous cell carcinoma in multiplanar (MPR) and three-dimensional (3D) images reconstructed from spiral CT using computer graphics and the importance of the computer software tools in permitting better visualization of the lesion. Two oral and maxillofacial radiologists and one neuroradiologist analysed the images. The usefulness of interactive reconstruction in the diagnosis, surgical planning and treatment evaluation, especially in the localization of the extent of these maxillofacial tumors, was demonstrated. Spiral CT-based computer graphics for displaying MPR and 3D images are an important adjunct in the evaluation of squamous cell carcinoma. PMID- 10455389 TI - Development of a compact computed tomographic apparatus for dental use. AB - OBJECTIVE: To describe a compact computed tomographic apparatus (Ortho-CT) for use in dental practice. METHODS: Ortho-CT is a cone-beam-type of CT apparatus consisting of a multifunctional maxillofacial imaging machine (Scanora, Soredex, Helsinki, Finland) in which the film is replaced with an X-ray imaging intensifier (Hamamatsu Photonics, Hamamatsu, Japan). The region of image reconstruction is a cylinder 32 mm in height and 38 mm in diameter and the voxel is a 0.136-mm cube. Scanning is at 85 kV and 10 mA with a 1 mm Cu filter. The scan time is 17 s comparable with that required for rotational panoramic radiography. A single scan collects 512 sets of projection data through 360 degrees and the image is reconstructed by a personal computer. The time required for image reconstruction is about 10 min. RESULTS: The resolution limit was about 2.0 lp mm-1 and the skin entrance dose 0.62 mGy. Excellent image quality was obtained with a tissue-equivalent skull phantom: roots, periodontal ligament space, lamina dura, and cancellous bone were clearly visualized. CONCLUSION: Ortho-CT provides three-dimensional images of excellent quality for dental use at a low entrance dose. PMID- 10455390 TI - Use of the direct magnification technique in dental radiology. AB - OBJECTIVE: To investigate the potential use of the direct magnification (DIMA) technique in dental radiology. METHODS: Radiographs of anatomical specimens and extracted teeth were prepared under clinical conditions using a microfocus (8 and 16 microns focal diameter) X-ray tube. Entrance dose was measured using an ionization chamber. Image quality was assessed in a single-blind test of pairs of images of extracted teeth with and without caries by ten observers. RESULTS: At 7.3 x magnification the values for exposure time and entrance dose were comparable with those for conventional intraoral radiography, 0.1 and 0.25 s and 1.5 and 2.9 mSv respectively. Diagnostic accuracy using the DIMA images was significantly (P < 0.0001) better compared with dental radiographs. CONCLUSION: DIMA images with significantly enhanced diagnostic accuracy can be obtained at comparable exposure times and entrance doses to conventional intraoral radiographs. PMID- 10455391 TI - Posterior lingual mandibular bone depression. PMID- 10455393 TI - The Buccal Object Rule. PMID- 10455394 TI - MRI: A conceptual overview. PMID- 10455392 TI - Ultrasonography-guided percutaneous drainage of cervicofacial infections. PMID- 10455395 TI - Molecular switches for regulating therapeutic genes. PMID- 10455396 TI - Coronary angioplasty: an end to early closing? PMID- 10455397 TI - Targeting endogenous platelet-derived growth factor B-chain by adenovirus mediated gene transfer potently inhibits in vivo smooth muscle proliferation after arterial injury. AB - Platelet-derived growth factor (PDGF), especially its B chain, has been implicated in the pathogenesis of vascular proliferative disorders such as atherosclerosis and restenosis after angioplasty. We constructed a replication deficient recombinant adenovirus containing the gene encoding the extracellular region of PDGF beta-receptor (PDGFXR) that binds PDGF-B chain and acts as its antagonist. The administration into balloon-injured rat carotid arteries of an adenovirus containing the Escherichia coli lacZ gene as a marker gene at 5 days after injury markedly facilitated efficacy of gene transfer, as compared with its administration immediately after injury. Adenovirus-mediated gene transfer of PDGFXR into injured arteries performed at 5 days resulted in a more than 50% reduction in the neointimal area of injured arteries at 14 days. In contrast, the administration of control adenoviruses containing lacZ gene or containing no foreign gene was without suppressive effects on neointima formation. The inhibition of neointima formation by the expression of PDGFXR was accompanied by a reduction in bromodeoxyuridine-labeled cells and nearly complete inhibition of tyrosine phosphorylation of both alpha- and beta-receptors for PDGF, but not of epidermal growth factor receptor, in injured arteries. This is the first report to indicate the usefulness of targeting a growth factor by expressing an extracellular binding region of a receptor using an adenovirus for the treatment of vascular proliferative disorders, and provide direct evidence that PDGF-B chain plays an essential role in neointimal formation. PMID- 10455398 TI - Improved gene transfer into canine hematopoietic repopulating cells using CD34 enriched marrow cells in combination with a gibbon ape leukemia virus-pseudotype retroviral vector. AB - We have used dogs to study gene transfer into hematopoietic stem cells, because of the applicability of results in dogs to human transplantation and the availability of canine disease models that mimic human diseases. Previously we reported successful gene transfer into canine marrow repopulating cells, however, gene transfer efficiency was low, usually below 0.1% (Kiem et al, Hum Gene Ther 1996; 7: 89). In this study we have used CD34-enriched marrow cells to study different retroviral pseudotypes for their ability to transduce canine hematopoietic repopulating cells. Cells were divided into two equal fractions that were cocultivated for 72 h with irradiated packaging cells producing vector with different retroviral pseudotypes (GALV, amphotropic or 10A1). The vectors used contained small sequence differences to allow differentiation of cells genetically marked by the different vectors. Nonadherent and adherent cells from the cultures were infused into four dogs after a myeloablative dose of 920 cGy total body irradiation. Polymerase chain reaction (PCR) analysis of DNA from peripheral blood and marrow after transplant showed that the highest gene transfer rates (up to 10%) were obtained with the GALV-pseudotype vector. Gene transfer levels have remained stable now for more than 18 months. Southern blot analysis confirmed the high gene transfer rate. Interference studies on canine D17 cells revealed that 10A1 virus behaved like an amphotropic virus and was not able to use the GALV receptor. In summary, our results show improved gene transfer into canine hematopoietic repopulating cells when CD34-enriched cells are transduced by cocultivation on a GALV-pseudotype packaging cell line in combination with a GALV-pseudotype vector. Furthermore, these results demonstrate that the monoclonal antibody to canine CD34 used in this study is able to enrich for hematopoietic repopulating cells. PMID- 10455399 TI - Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. AB - Conventional methods for rAAV purification that are based on cesium chloride ultracentrifugation have often produced vector preparations of variable quality and resulted in significant loss of particle infectivity. We report here several novel purification strategies that involve the use of non-ionic iodixanol gradients followed by ion exchange or heparin affinity chromatography by either conventional or HPLC columns. These methods result in more than 50% recovery of rAAV from a crude lysate and routinely produce vector that is more than 99% pure. More importantly, the new purification procedures consistently produce rAAV stocks with particle-to-infectivity ratios of less than 100, which is significantly better than conventional methods. The new protocol increases the overall yield of infectious rAAV by at least 10-fold and allows for the complete purification of rAAV in 1 working day. Several of these methods should also be useful for large-scale production. PMID- 10455400 TI - High-titer recombinant adeno-associated virus production utilizing a recombinant herpes simplex virus type I vector expressing AAV-2 Rep and Cap. AB - Recombinant adeno-associated virus type 2 (rAAV) vectors have recently been used to achieve long-term, high level transduction in vivo. Further development of rAAV vectors for clinical use requires significant technological improvements in large-scale vector production. In order to facilitate the production of rAAV vectors, a recombinant herpes simplex virus type I vector (rHSV-1) which does not produce ICP27, has been engineered to express the AAV-2 rep and cap genes. The optimal dose of this vector, d27.1-rc, for AAV production has been determined and results in a yield of 380 expression units (EU) of AAV-GFP produced from 293 cells following transfection with AAV-GFP plasmid DNA. In addition, d27.1-rc was also efficient at producing rAAV from cell lines that have an integrated AAV-GFP provirus. Up to 480 EU/cell of AAV-GFP could be produced from the cell line GFP 92, a proviral, 293 derived cell line. Effective amplification of rAAV vectors introduced into 293 cells by infection was also demonstrated. Passage of rAAV with d27. 1-rc results in up to 200-fold amplification of AAV-GFP with each passage after coinfection of the vectors. Efficient, large-scale production (>109 cells) of AAV-GFP from a proviral cell line was also achieved and these stocks were free of replication-competent AAV. The described rHSV-1 vector provides a novel, simple and flexible way to introduce the AAV-2 rep and cap genes and helper virus functions required to produce high-titer rAAV preparations from any rAAV proviral construct. The efficiency and potential for scalable delivery of d27.1-rc to producer cell cultures should facilitate the production of sufficient quantities of rAAV vectors for clinical application. PMID- 10455401 TI - Liposome-mediated NGF gene transfection following neuronal injury: potential therapeutic applications. AB - We have systematically investigated the therapeutic potential of cationic liposome-mediated neurotrophic gene transfer for treatment of CNS injury. Following determination of optimal transfection conditions, we examined the effects of dimethylaminoethane-carbamoyl-cholesterol (DC-Chol) liposome-mediated NGF cDNA transfection in injured and uninjured primary septo-hippocampal cell cultures and rat brains. In in vitro studies, we detected an increase of NGF mRNA in cultures 1 day after transfection. Subsequent ELISA and PC12 cell biological assays confirmed that cultured cells secreted soluble active NGF into the media from day 2 after gene transfection. Further experiments showed that such NGF gene transfection reduced the loss of chol- ine acetyltransferase (ChAT) activity in cultures following calcium-dependent depolarization injury. In in vivo studies, following intraventricular injections of NGF cDNA complexed with DC-Chol liposomes, ELISA detected nine- to 12-fold increases of NGF in rat CSF. Further studies showed that liposome/NGF cDNA complexes could attenuate the loss of cholinergic neuronal immunostaining in the rat septum after traumatic brain injury (TBI). Since deficits in cholinergic neurotransmission are a major consequence of TBI, our studies demonstrate for the first time that DC-Chol liposome-mediated NGF gene transfection may have therapeutic potential for treatment of brain injury. PMID- 10455403 TI - IGF-I gene transfer in thermally injured rats. AB - Exogenous insulin-like growth factor-I (IGF-I) is known to improve the pathophysiology of a thermal injury, however, deleterious side-effects have limited its utility. Cholesterol-containing cationic liposomes that encapsulate complementary DNA (cDNA) are nonviral carriers used for in vivo gene transfection. We propose that liposome IGF-I gene transfer will accelerate wound healing in burned rats and attenuate deleterious side-effects associated with high levels of IGF-I. To test this hypothesis IGF-I gene constructs, encapsulated in liposomes, were studied for their efficacy in modulating the thermal injury response. Thirty adult male Sprague-Dawley rats were given a 60% TBSA scald burn and randomly divided into three groups to receive weekly subcutaneous injections of liposomes plus the lacZ gene coding for beta-galactosidase, liposomes plus cDNA for IGF-I and beta-galactosidase or liposomes plus the rhIGF-I protein. Body weights and wound healing were measured. Muscle and liver dry/wet weights and IGF I concentrations in serum, skin and liver were measured by radioimmunoassay. Transfection was confirmed by histochemical staining for beta-galactosidase. Rats receiving the IGF-I cDNA constructs exhibited the most rapid wound re epithelialization and greatest increase in body weight and gastrocnemius muscle protein content (P < 0.05). Local IGF-I protein concentrations in the skin were higher when compared to liposomes containing only the lacZ gene (P < 0.05) Transfection was apparent in the cytoplasm of myofibroblasts, endothelial cells and macrophages of the granulation tissue. Liposomes containing the IGF-I gene constructs proved effective in preventing muscle protein wasting and preserving total body weight after a severe thermal injury. PMID- 10455402 TI - Cell-specific nuclear import of plasmid DNA. AB - One factor limiting the success of non-viral gene therapy vectors is the relative inability to target genes specifically to a desired cell type. To address this limitation, we have begun to develop cell-specific vectors whose specificity is at the level of the nuclear import of the plasmid DNA. We have recently shown that nuclear import of plasmid DNA is a sequence-specific event, requiring the SV40 enhancer, a region known to bind to a number of general transcription factors (Dean DA, Exp Cell Res 1997; 230: 293). From these studies we developed a model whereby transcription factor(s) bind to the DNA in the cytoplasm to create a protein-DNA complex that can enter the nucleus using the protein import machinery. Our model predicts that by using DNA elements containing binding sites for transcription factors expressed in unique cell types, we should be able to create plasmids that target to the nucleus in a cell-specific manner. Using the promoter from the smooth muscle gamma actin (SMGA) gene whose expression is limited to smooth muscle cells, we have created a series of reporter plasmids that are expressed selectively in smooth muscle cells. Moreover, when injected into the cytoplasm, plasmids containing portions of the SMGA promoter localize to the nucleus of smooth muscle cells, but remain cytoplasmic in fibroblasts and CV1 cells. In contrast, a similar plasmid carrying the SV40 enhancer is transported into the nuclei of all cell types tested. Nuclear import of the SMGA promoter containing plasmids could be achieved when the smooth muscle specific transcription factor SRF was expressed in stably transfected CV1 cells, supporting our model for the nuclear import of plasmids. Finally, these nuclear targeting sequences were also able to promote increased gene expression in liposome- and polycation-transfected non-dividing cells in a cell-specific manner, similar to their nuclear import activity. These results provide proof of principle for the development of cell-specific non-viral vectors for any desired cell type. PMID- 10455404 TI - Differential expression and secretion of alpha 1 anti-trypsin between direct DNA injection and implantation of transfected myoblast. AB - Muscle can be used for systemic delivery of non-muscle proteins. In order to investigate the relative effectiveness of direct DNA plasmid injection versus implantation of genetically modified myogenic cell lines, we have used the human alpha 1 anti-trypsin (alpha1AT) cDNA driven by either cytomegalovirus (CMV) or the muscle creatine kinase 3.3 kb (MCK) promoter in immunodeficient mice. We demonstrate that the implantation of transfected myoblasts stably expressing the human alpha1AT cDNA generates a more persistent production of alpha1AT than does direct intramuscular injection of the same construct as plasmid DNA. Moreover, immunohistological labelling of muscle sections implanted with myoblasts show that the newly formed muscle fibres are those containing the human protein. PMID- 10455405 TI - Therapeutic efficacy of the thymidine kinase/ganciclovir system on large experimental gliomas: a nuclear magnetic resonance imaging study. AB - Contradictory experimental results and human trials have questioned the clinical relevance of the HSVtk/ganciclovir system. To bypass the problem of transfection efficiency, we used a glioma cell line stably expressing the HSVtk gene, which was also fully characterized from gene to protein. We also designed a more clinically relevant experimental protocol, consisting of late GCV delivery on large tumor formations. In short-term studies, histological examination revealed a significant decrease in tumor volume in GCV-treated animals from day 1 or from day 10 after cell inoculation. We observed that late GCV delivery is as efficient as early delivery, probably because GCV can reach tumor cells more easily when neoangiogenesis occurs. In long-term experiments, the survival of treated rats bearing 15-day tumors was improved by 60% compared with C6 control animals. Surprisingly, a 30% survival rate was observed in C6TK control animals. Nuclear magnetic resonance imaging demonstrated, in all surviving animals, a complete regression of tumors without mass effect. These results clearly demonstrate that the HSVtk/GCV system remains a potent therapeutic strategy, even when tested in large tumors, in contrast with the microscopic tumor formations previously reported. PMID- 10455406 TI - Development of a modified selective amplifier gene for hematopoietic stem cell gene therapy. AB - We have proposed a novel concept, ie selective expansion of transduced cells, to overcome the low efficiency of gene transfer into hematopoietic stem cells. Previously, a fusion gene encoding a chimeric receptor (DeltaGCRER) between the mouse granulocyte colony-stimulating factor receptor (G-CSFR) and the hormone binding domain of rat estrogen receptor was constructed as a 'selective amplifier gene'. Although the chimeric gene conferred estrogen-inducible proliferation on the transduced Ba/F3 cells, it also mediated differentiation of the retrovirally transduced 32D cells upon estrogen treatment. Since only a growth signal is required for our purpose, we further modified the DeltaGCRER gene to attenuate its differentiation signal. Based on the observation that tyrosine-703 in wild type G-CSFR plays a pivotal role in transmitting the differentiation signal, phenylalanine was substituted for this residue in DeltaGCRER. When the resultant selective amplifier gene (DeltaY703F-GCRER gene) was expressed in 32D cells, sustained growth was supported by estrogen, while differentiation was suppressed. These cells ceased to grow upon estrogen withdrawal and differentiated with G-CSF treatment. The present findings suggested that DeltaY703F-GCRER may have desirable properties as a selective amplifier for hematopoietic stem cell expansion and gene therapy. PMID- 10455407 TI - Cellular contaminants of adeno-associated virus vector stocks can enhance transduction. AB - Transduction efficiency of different types of recombinant (r)AAV-2 based vectors preparations markedly differed, with apparently no correlation with the replicative titers. Using HeLa cells as target for transduction, 105 and 30 infectious units were necessary to observe one transductant using respectively cesium-chloride-purified rAAV and crude lysates of producer cells obtained by sonication. The purified vectors were however able to transduce HEK-193 cells efficiently, but transgene expression was detected with some delay compared with crude lysates. The unexpected high transduction efficiency of sonicated crude lysates was due to virally mediated gene transfer, since similar sonicated crude lysates, but with no AAV rep and cap genes, did not lead to detection of transgene products after incubation with HeLa cells. Furthermore, sonicated cellular extracts of 293 or 293/T cells given in trans stimulate transduction of HeLa cells by purified rAAV. In contrast, neither extracts from the adenovirus E1 transformed 911 cell line, nor from other cell lines not harboring any adenovirus gene, had enhancing effect on rAAV-mediated transduction. These data suggest that 293 sonicated extracts contain factors which stimulate rAAV-mediated transduction of cells that are normally poorly transduced and offer a system to identify such factors and to characterize further the steps limiting the transfer of gene by AAV vectors. PMID- 10455408 TI - Induction of apoptosis and G2/M arrest by infection with replication-deficient adenovirus at high multiplicity of infection. AB - Replication-deficient adenoviruses are among the most widely used vectors in gene therapy and are also becoming increasingly popular as analytical tools in basic research. However, significant toxicity of these vectors in vivo has been reported. Here, we show that in an in vitro setting, first generation adenoviruses lead to growth retardation, prolongation of the G2/M phase and induction of apoptosis if applied at a high multiplicity of infection (MOI). These findings were obtained in p53-deficient hepatocytes, derived from knock-out mice (A2 cells) and in several tumor cell lines containing wild-type (wt) or mutant p53. Apoptosis induction was correlated with increased levels of p53 and bax proteins and it was stronger in cells containing wt p53 as compared with cells lacking functional p53. Apoptosis was highly dependent on the MOI used with marked effects starting at an MOI twice as high as needed for 100% gene transfer. Expression of the adenoviral E4 ORF6 gene as well as adenoviral replication were detected in all cell lines infected with first generation adenovirus. Apoptosis could be considerably reduced but not abrogated by UV inactivation of adenovirus, which indicates proapoptotic effects caused by the infection event as well as by residual adenoviral gene expression or adenoviral replication. First generation adenoviruses apparently display proapoptotic activity if used at higher MOIs, which may be of relevance when these vectors are used as analytical or gene therapeutic tools. PMID- 10455410 TI - Induction of complement attack on human cells by Gal(alpha1,3)Gal xenoantigen expression as a gene therapy approach to cancer. AB - Galactose(alpha1,3)galactose on the surface of cells of non-primate organs is the major xenoantigen responsible for hyperacute rejection in xenotransplantation. The antigen is synthesised by (alpha1, 3)galactosyl transferase. Humans lack this enzyme and their serum contains high levels of pre-existing natural antibody which recognises the structure and activates complement. We have evaluated in vitro the potential for delivery of this enzyme to sensitise human cells to complement attack as a gene therapy approach to cancer. Retrovirus-mediated delivery of (alpha1,3)galactosyl transferase resulted in high level expression which led to serum-mediated lysis of five human cell targets, including endothelial and primary melanoma cells. Lysis was specific for those cells expressing the antigen in a mixed cell population. The mechanism of cell lysis mimicked that involved in hyperacute rejection: activation of the classical complement pathway by natural antibody specific for galactose(alpha1,3)galactose. The degree of lysis was determined by both the level of specific antibody and the expression of glycophosphatidylinositol-linked complement regulatory proteins. We conclude that expression of (alpha1,3)galactosyl transferase is a promising new therapeutic approach for cancer gene therapy, avoiding toxicity problems associated with application of prodrugs and with the potential to elicit further immunological responses. PMID- 10455409 TI - Increase of BCNU sensitivity by wt-p53 gene therapy in glioblastoma lines depends on the administration schedule. AB - In this article, we investigated the effect induced by the reintroduction of wild type p53 (wt-p53) protein on BCNU sensitivity in the ADF glioblastoma line. Using a wt-p53 recombinant adenovirus (Ad-p53), we demonstrated that exogenous wt-p53 expression was able to increase the sensitivity to BCNU in ADF cells. Interestingly, this effect was more evident when Ad-p53 infection was performed after BCNU treatment compared with the opposite sequence. To understand the biological basis of these different behaviors, we analyzed the cell cycle of the differently treated cells. We found that Ad-p53 infection induced a persistent accumulation of cells in the G0/G1 phase while, as expected, BCNU induced a block in the G2-M phase. Ad-p53-->BCNU sequence did not significantly modify the cell cycle profile in respect of Ad-p53 infected cells. In contrast, BCNU-->Ad-p53 sequence provoked G2-M arrest similar to that observed after treatment with BCNU alone, but prevented the later recovery of the cells through the cell cycle, by driving the cells to apoptotic death. These results demonstrate that the administration sequence is important to increase drug sensitivity. To generalize the phenomenon observed on ADF line, the antiproliferative effect of the two different schedules was analyzed on other glioblastoma lines (A172, CRS-A2, U373MG) with different BCNU sensitivity and p53 status. The data obtained confirm that the wt-p53 gene transfer enhances BCNU sensitivity in glioblastoma cells depending on the administration sequence. PMID- 10455411 TI - Retroviral transduction of quiescent haematopoietic cells using a packaging cell line expressing the membrane-bound form of stem cell factor. AB - Gene therapy vectors based on murine retroviruses are unable to transduce non dividing cells. This has proven a particular problem in the haematopoietic system where the target cells of choice, the pluripotent stem cells are quiescent. In an attempt to circumvent this difficulty we have constructed a retroviral producer line that expresses the membrane bound form of human recombinant stem cell factor (SCF) on its cell surface. This should enable the retroviral producers to deliver a growth signal to the target cells simultaneous with their exposure to retrovirus. We tested the ability of these modified producers to transduce a growth factor-starved, SCF-dependent cell line (TF-1) and demon- strated that these cells, though quiescent, can still be successfully transduced. This approach was extended to targeting of umbilical cord blood CD34+ cells, a predominantly quiescent population that normally require the addition of cytokines for efficient transduction. Using the SCF-expressing producer line in the absence of exogenously added cytokines, we observed a marked stimulation in transduction efficiency over that achieved using the parent producer line alone. Colonies derived from these cells arising in semi-solid media were also shown to be positive for expression of a retrovirally encoded reporter gene. PMID- 10455412 TI - Overexpression of urokinase receptor in mammalian cells following administration of the in vitro transcribed encoding mRNA. AB - The ability to overexpress physiologically important proteins in cultured mammalian cells after delivering the encoding mRNAs could have important applications for analyzing their in vivo functions. To explore the potential of this approach, urokinase-type plasminogen activator receptor (uPAR), a membrane protein extensively modified post-translationally, was selected. The uPAR encoding mRNAs, containing different 5' and 3' untranslated regions (UTR) were tested in cultured human osteosarcoma (HOS) cells following a cationic lipid mediated delivery. The most effective structure was the capped and polyadenylated transcript containing Xenopus beta-globin 5' and 3' UTRs. Delivering this mRNA to HOS cells resulted in a significant increase of uPAR expression in 89% of the cells, measured by flow cytometry. Using a radioligand binding assay, the increase in functional uPAR levels was found to be up eight- to 11-fold between 8 and 48 h and up three-fold at 72 h after delivery. A similar increase in uPAR levels was achievable in a number of mammalian cell lines. Surprisingly, poly(A) tailed mRNA leading to a uPAR production highest in magnitude and duration did not demonstrate increased intracellular stability compared with other tested mRNAs. Thus, the exceptional translational performance is not likely the result of an increased mRNA half-life. These results demonstrate that, after delivery of selected mRNAs into mammalian cells, immediate and significant overexpression of a post-translationally modified protein is achievable. PMID- 10455413 TI - 'Autoreplication' of the vector genome in recombinant adenoviral vectors with different E1 region deletions and transgenes. AB - High transgene stabilities of 1 year and more have been reported in immunodeficient hosts after adenovirus-mediated gene transfer. Transgene persistence of this duration could be due to inherently high stability of the episomal viral vector DNA. An alternative explanation would be limited 'autoreplication' of transgenic vector DNA, just sufficient to counteract slow but continuous degradation within the host cells. Autoreplication could occur in the absence of any production of infectious virus particles, based on residual activity of the adenoviral DNA replication system only. To test this hypothesis, a series of DNA metabolic labeling studies in non-permissive cells cultures transfected with different vectors was conducted. Due to extensive E1 region deletions none of the vectors was able to produce viral progeny in non-permissive cells. Vectors fell into two categories, however, with respect to their autoreplication potential. Neosynthesis of vector DNA in non-permissive vector transfected cells was readily detectable in 'type A', but not in 'type B' vectors. In addition to their different transgene expression cassettes, vector DNA sequencing showed a less extensive E1 deletion in type A (nucleotides 453 3333 of wild-type virus) as compared to type B vectors (nucleotides 325-3523). Autoreplication was also associated with high transcriptional activity of several viral genes (E1B-14k, adenoviral DNA polymerase, single-strand DNA-binding protein, E4-25k), in contrast to type B vectors. In addition to these 'wild-type' transcripts, 'irregular' recombinant transcripts were detected in autoreplication vectors which contained the transgenic cDNA in conjunction with adenoviral vector sequences. Exogenous or cryptic promotors may (under certain conditions) enhance the transcriptional activity of a vector in such a way that autoreplication occurs. Conditions determining the level of transcriptional enhancement (extent of E1 deletion, type of promoter and transgene, etc) need to be further defined before rational design of adenovectors with high autoreplication capacity becomes possible. In summary, we have shown autoreplication to be a novel feature of certain E1-deleted adenovectors with likely relevance for their stability in vivo, but also with possibly adverse consequences for target cell function or vector immunogenicity. Full characterization of adenoviral vector systems should therefore include a description of their autoreplication capacity. PMID- 10455414 TI - The sCYMV1 hairpin ribozyme: targeting rules and cleavage of heterologous RNA. AB - The catalytic center of the RNA from the negative strand of the satellite RNA of chicory yellow mottle virus type 1 (sCYMV1) is in the hairpin ribozyme family, has catalytic activity, and cleaves substrates before a preferred GUA sequence. This is different from that of the satellite RNA from the negative strand of tobacco ringspot virus (sTRSV) which prefers a GUC sequence at the site of cleavage. The sCYMV1 hairpin ribozyme has now been developed for cleaving heterologous RNA substrates. When helix 1 was extended from the native 5 bp to 6 bp with a newly added A:U base pair, catalytic activity increased three-fold. The preferred sequence for the substrate loop was the native A*GUA sequence where * is the site of cleavage. When each nucleotide in this sequence was changed to each of the other three nucleotides, catalytic activity decreased 66-100%. RNA targets, containing this A*GUA sequence, were located in both human papillomavirus and HIV-1. Ribozymes were developed which efficiently cleaved these targets in vitro. These results identify a new class of hairpin ribozymes capable of cleaving substrates before a preferred GUA sequence rather than the GUC preferred by the sTRSV hairpin ribozyme. This expands the repertoire of target sites available for gene therapy using the hairpin ribozyme. PMID- 10455415 TI - Augmentation of local antitumor immunity in the liver by tumor vaccine modified to secrete murine interleukin 12. AB - Minimal residual lesions have been a major problem in surgical management of cancer. We transfected M5076 with murine IL-12 gene by a retroviral vector, established a stable transfectant secreting IL-12 and investigated its antitumor effects on a spontaneous liver metastasis murine model of M5076 reticulum cell sarcoma. Subcutaneous vaccination of the irradiated transfectant into the remote skin following the amputation of the tumor-bearing limb improved survival when compared with the vaccination of irradiated parental cells (control). Cytotoxic activities against parental M5076 were significantly stronger in the hepatic lymphocytes from the mice vaccinated with the IL-12 transfectant than those from the control. IFN-gamma production of hepatic lymphocytes when they were cocultured with the parental cells was significantly augmented in mice vaccinated with the IL-12 transfectant compared with the control. On the other hand, both cytotoxic activity and IFN-gamma production of spleen cells in the M5076 vaccinated and transfectant-vaccinated mice were at similar levels. Immunophenotypic analysis revealed the selective increase of CD3+NK1+ population in the liver from the transfectant-vaccinated mice. These results suggest that tumor vaccines genetically modified to secrete IL-12 continuously at a relatively low level preferentially augment local antitumor activity in the liver rather than systemic immune responses. This strategy warrants further investigation as an adjuvant modality in the management of postoperative residual tumors. PMID- 10455416 TI - Augmentation of natural immunity to a pro-inflammatory cytokine (TNF-alpha) by targeted DNA vaccine confers long-lasting resistance to experimental autoimmune encephalomyelitis. AB - TNF-alpha is thought to be a key pro-inflammatory cytokine in T cell-mediated autoimmune diseases, particularly in rheumatoid arthritis (RA) and multiple sclerosis (MS). Experimental autoimmune encephalomyelitis (EAE) serves as an animal model for MS. The current study observes a notable TNF-alpha-specific antibody titer generated during the course of EAE, apparently not sufficient to prevent the development of disease. Administration of TNF-alpha-naked DNA vaccine enhanced the production of TNF-alpha-specific antibody titer and conferred EAE resistance. These antibodies were found to be neutralizing in vitro and capable of inhibiting the development of disease when transferred to other EAE rats. Thus, modulation of EAE with TNF-alpha DNA vaccines enhances the regulation of natural immunity to a self pro-inflammatory cytokine and provides a tool by which the immune system is encouraged to elicit anti-self protective immunity to restrain its own harmful reactivity when such a response is needed. PMID- 10455417 TI - Ganciclovir-mediated in vivo elimination of myeloid leukemic cells expressing the HSVtk gene induces HSVtk loss variants. AB - The in vivo elimination of suicide gene-expressing tumor cells with prodrug treatment can induce protective immunity against wild-type tumors. In this study, we determined the efficacy and safety of the in vivo elimination of HSVtk expressing cells with ganciclovir treatment of a bystander cell killing insensitive leukemic cell line. The retroviral construct pLTk+NeoDeltaMo, containing the HSVtk gene and the NeoR gene in a bicistronic unit, was introduced into rat leukemic LT12 cells. LT12/Tk+N cells showed a 1000- to 10 000-fold increased sensitivity to ganciclovir in vitro. In vitro mixing experiments demonstrated that LT12 cells were not susceptible to bystander cell lysis by LT12/Tk+N-2 cells exposed to ganciclovir. Rats injected s.c. with cloned LT12/Tk+N-2 cells developed tumors reaching a diameter of 3-4 cm after 40 days. Rats treated with gan- ciclovir twice daily for 5 consecutive days starting at day 7 did not develop s.c. tumors. Large established s.c. LT12/Tk+N-2 tumors completely regressed after ganciclovir treatment. However, recurrences of s.c. tumors were observed that were no longer sensitive to ganciclovir treatment. In vitro analysis of aspirates from the recurrent tumors demonstrated loss of HSVtk expression. In vitro culture of LT12/Tk+N-2 cells in soft agar in the presence of ganciclovir indicated that the frequency with which HSVtk-loss variants occurred is approximately one per 104 cells. The in vivo occurrence of HSVtk-loss variants escaping ganciclovir-induced elimination may have important implications for vaccination protocols using HSVtk gene expressing tumor cells that are not susceptible to bystander cell killing. PMID- 10455418 TI - Noradrenaline transporter gene transfer for radiation cell kill by 131I meta iodobenzylguanidine. AB - Meta-iodobenzylguanidine conjugated to 131I-iodine is an effective agent for the targeted radiotherapy of tumors of neural crest origin which express the noradrenaline transporter (NAT). The therapeutic application of 131I MIBG is presently limited to the treatment of phaeochromocytoma, neuroblastoma, carcinoid and medullary thyroid carcinoma. To determine the feasibility of MIBG targeting for a wider range of tumor types, we employed plasmid-mediated transfer of the NAT gene into a human glioblastoma cell line (UVW) which does not express the NAT gene. This resulted in a 15-fold increase in uptake of MIBG by the host cells. A dose-dependent toxicity of 131I MIBG to the transfectants was demonstrated using three methods: (1) survival of clonogens derived from monolayer culture; (2) survival of clonogens derived from disaggregated multicellular spheroids; and (3) spheroid growth delay. 131I MIBG was twice as toxic to cells in spheroids compared with those in monolayers, consistent with a greater effect of radiation cross-fire (radiological bystander effect) from 131I beta-radiation in the three dimensional tumor spheroids. The highest concentration of 131I MIBG tested (1 MBq/ml) was nontoxic to UVW control cells or spheroids transfected with the NAT gene in reverse orientation. These findings are encouraging for the development of NAT gene transfer-mediated 131I MIBG therapy. PMID- 10455419 TI - A novel endothelial cell-based gene therapy platform for the in vivo delivery of apolipoprotein E. AB - A major focus in gene therapy has been the use of recombinant viruses to deliver genes in vivo. Although this approach shows much promise, there are many safety concerns associated with the use of viral materials in the treatment of human diseases. Our alternative cell-based gene therapy approach utilizes endothelial cells (Pro 175) isolated from the murine embryonic yolk sac. These endothelial cells were evaluated for their potential use in gene therapy as a gene delivery platform. As a test model, we used these cells to deliver apolipoprotein E (apoE) in the murine apoE knockout atherosclerosis model. The lack of apoE protein in these animals results in high levels of serum cholesterol and formation of severe aortic plaques and lesions at a young age. After transplantation of the apoE secreting Pro 175 endothelial cells into apoE-deficient mice, serum cholesterol levels were measured at 2 week intervals. During the 3 months after the initiation of these experiments, levels of cholesterol in the animals having received the apoE secreting endothelial cells were statistically lower compared with the levels of age-matched controls having received non-secreting endothelial cells. Concomitant with cholesterol reduction, atherosclerotic aortic plaques were noticeably reduced in the experimental apoE+ animals. These results highlight the potential of these unique endothelial cells as an efficient delivery platform for somatic gene therapy. PMID- 10455420 TI - Lentivirus-mediated Bcl-2 expression in betaTC-tet cells improves resistance to hypoxia and cytokine-induced apoptosis while preserving in vitro and in vivo control of insulin secretion. AB - betaTC-tet cells are conditionally immortalized pancreatic beta cells which can confer long-term correction of hyperglycemia when transplanted in syngeneic streptozocin diabetic mice. The use of these cells for control of type I diabetes in humans will require their encapsulation and transplantation in non-native sites where relative hypoxia and cytokines may threaten their survival. In this study we genetically engineered betaTC-tet cells with the anti-apoptotic gene Bcl 2 using new lentiviral vectors and showed that it protected this cell line against apoptosis induced by hypoxia, staurosporine and a mixture of cytokines (IL-1beta, IFN-gamma and TNF-alpha). We further demonstrated that Bcl-2 expression permitted growth at higher cell density and with shorter doubling time. Expression of Bcl-2, however, did not inter- fere either with the intrinsic mechanism of growth arrest present in the betaTC-tet cells or with their normal glucose dose-dependent insulin secretory activity. Furthermore, Bcl-2 expressing betaTC-tet cells retained their capacity to secrete insulin under mild hypoxia. Finally, transplantation of these cells under the kidney capsule of streptozocin diabetic C3H mice corrected hyperglycemia for several months. These results demonstrate that the murine betaTC-tet cell line can be genetically modified to improve its resistance against different stress-induced apoptosis while preserving its normal physiological function. These modified cells represent an improved source for cell transplantation therapy of type I diabetes. PMID- 10455421 TI - Retrovirus-mediated WASP gene transfer corrects defective actin polymerization in B cell lines from Wiskott-Aldrich syndrome patients carrying 'null' mutations. AB - Boys affected with Wiskott-Aldrich syndrome (WAS) present with variable association of thrombocytopenia, eczema and immune deficiency. If untreated, WAS patients may succumb to intracerebral hemorrhages, severe infections or malignancies. Allogeneic bone marrow transplantation (BMT) can cure all aspects of the disease, but HLA-identical donors are not available to all patients and mismatched BMTs are unfortunately associated with high mortality and morbidity. The good success of HLA-matched BMT, however, makes WAS a potential candidate for hematopoietic stem cell gene therapy. WAS patients carry mutations of the Wiskott Aldrich syndrome protein gene encoding WASP, a 502-amino acid proline-rich protein with demonstrated involvement in the organization of the actin cytoskeleton. To verify the feasibility of genetic correction for this disease, the WASP cDNA was expressed in EBV-immortalized B cell lines obtained from WAS patients using a retroviral vector. Transduced WAS cells showed levels of WASP expression similar to those found in cells from normal donors, without detectable effects on viability or growth characteristics. In addition, retrovirus-mediated expression of WASP led to improvement of cytoplasmic F-actin expression and formation of F-actin-positive microvilli, a process shown to be defective in untransduced WAS cell lines. These preliminary results indicate a potential use for retrovirus-mediated gene transfer as therapy for WAS. PMID- 10455422 TI - Elimination of lysosomal storage in brains of MPS VII mice treated by intrathecal administration of an adeno-associated virus vector. AB - Mucopolysaccharidosis type VII (MPS VII) is an inherited lysosomal storage disease caused by insufficient beta-glucuronidase (GUS). To provide gene therapy in a mutant mouse model of this disease, we have used a recombinant adeno associated virus (rAAV) vector to deliver GUS cDNA to a variety of tissues. Although intravenous administration of vector produced therapeutic levels of GUS in the liver, delivery to the brain was inadequate. To improve delivery to the brain intrathecal injection of the vector into the cerebrospinal fluid was employed. This route of administration to either neonatal or adult mutant mice resulted in therapeutic levels of GUS in the brain and the elimination of storage granules in brain tissue. PMID- 10455423 TI - Cationic lipids are essential for gene delivery mediated by intravenous administration of lipoplexes. AB - It was recently suggested that intravenously administered lipoplexes serve as a depot for the extracellular release of naked DNA and it is the naked DNA that mediates gene delivery in the lung. If this is the mechanism responsible for gene expression, we reasoned that continuous infusion of plasmid DNA should also result in significant lung expression in the absence of lipoplexes. Moreover, the infusion of non-coding plasmid DNA should inhibit gene delivery by lipoplexes. Infusion of plasmid DNA at a rate of 80 microg/min into the tail vein of a mouse resulted in a DNA serum concentration of 800 microg/ml. This was equivalent to a transcriptionally active DNA concentration of 120 microg/ml plasma as determined by an in vitro transfection assay. In spite of this high level of transcriptionally active DNA, there was no significant gene expression in the lung or any other organ tested. In addition, when lipoplex containing a reporter gene was injected, followed by an infusion of non-coding plasmid DNA as a potential competing molecule for DNA released from the lipoplex there was no effect on gene expression. These experiments indicate that the cationic lipid component of the lipoplex functions in an active capacity beyond that of a simple passive release matrix for plasmid DNA. PMID- 10455424 TI - Tissue-selective expression of dominant-negative proteins for the regulation of vascular smooth muscle cell proliferation. AB - The transcription factors c-myb and c-myc are essential for vascular smooth muscle cell (VSMC) replication and are rapidly induced following mitogenic stimulation of quiescent VSMCs in vitro and in vivo following balloon catheter injury. Consequently, interference with c-myb and c-myc function provides a possible avenue for the prevention of VSMC proliferation associated with intimal hyperplasia. We have carried out studies focused on the inhibition of VSMC proliferation using dominant-negative gene constructs incorporating the DNA binding domains of the c-myb or c-myc genes fused to the repressor domain of the Drosophila engrailed gene. Transient transfection of rat, rabbit and human vascular SMCs results in a dramatic inhibition of proliferation for at least 72 h after transfection. Furthermore, this inhibition of cellular proliferation was found to be due, at least in part, to the induction of apoptosis. Coupling expression of the chimeric dominant-negative proteins to transcriptional regulatory elements of the human vascular smooth muscle alpha-actin gene allows specific targeting of vascular smooth muscle cells. PMID- 10455426 TI - Gene therapy for the millenium: in with the new PMID- 10455427 TI - Fetal gene therapy. PMID- 10455425 TI - Insulation of a conditionally expressed transgene in an adenoviral vector. AB - Replication-defective recombinant adenoviruses provide an efficient system for in vivo gene transfer and numerous studies have demonstrated that this vector can accommodate tissue-specific promoters to restrict the expression of a transgene to a particular subset of cells. However, in some cases the selectivity of expression is lost when the tissue-specific promoter is placed in an adenoviral environment. In an attempt to restore the conditionality of expression of the transgene driven by the human ERBB2 promoter, we have flanked the expression cassette in 5' and 3' orientations with a 250 bp sequence containing the bovine growth hormone transcriptional stop signal for cloning into a recombinant adenovirus. The data presented here clearly demonstrate that these 'insulator' elements are able to restrict the expression of the transgene (herpes simplex thymidine kinase) to ERBB2-expressing cells and therefore to restore the selectivity mediated by the ERBB2 promoter. This approach could be generally useful to insulate expression cassettes in adenoviral vectors. PMID- 10455428 TI - A role for intracellular immunization in chemosensitization of tumor cells? AB - Acquired drug resistance represents a major cause of chemotherapy failure in patients with cancer. The characterization of the molecular pathways involved in drug resistance has provided us with new targets to overcome this problem. Many of these target proteins are often overexpressed in human cancers. A number of gene therapy strategies, including antisense oligonucleotides, ribozymes and single-chain antibodies, have been developed to achieve the selective modulation and inhibition of various cellu- lar proteins. Thus, these approaches can be exploited to modulate the resistance phenotype of tumor cells. These gene therapy strategies represent a novel and unique way to enhance the sensitivity of tumor cells to chemotherapeutic drugs. This review will focus on the use of intracellular immunization as a means to modulate the expression of specific genetic determinants involved in the drug resistance phenotype. PMID- 10455429 TI - Angiogenesis induced in muscle by a recombinant adenovirus expressing functional isoforms of basic fibroblast growth factor. AB - The present work studies the effects of a replication-deficient adenovirus (Ad), Ad-RSVbFGF, bearing the human basic fibroblast growth factor (bFGF) cDNA, as a potential vector for therapeutic angiogenesis of ischemic diseases. The different isoforms of the protein were expressed from the viral vector in various cell types and, although the cytoplasmic isoform does not possess a signal peptide, we observed its release from a muscle cell line. The proteins were fully functional when tested in a long-term survival assay of quiescent fibroblasts. After endothelial cell infection with Ad-RSVbFGF, we observed an 80&percnt increase in the mean length of the capillary-like tubes that differentiated in a three dimensional model of angiogenesis. We evaluated angiogenesis directly in mice 14 days after subcutaneous injection of Matrigel plugs containing Ad-RSVbFGF. A marked neovascularization was observed in the Matrigel plugs and in the surrounding tissues. Finally, the recombinant virus was injected into the hindlimb muscles of mdx mice. A 2.5-fold increase in bFGF content of the muscle was observed 6 days after injection, without any significant variations detected in the animal sera. Immunohistological detection showed an increased number of large-caliber vessels in the treated muscles as compared with control muscles. These results demonstrate that Ad-mediated transfer of the human bFGF gene can induce angiogenesis in muscle, making this tissue a potential target for the treatment of ischemic diseases. PMID- 10455430 TI - Bicistronic retroviral vectors for combining myeloprotection with cell-surface marking. AB - We have developed a retroviral vector coexpressing the multidrug-resistance 1 (MDR1) cDNA for inducing cancer drug resistance and the truncated version of the low-affinity nerve growth factor receptor (DeltaLNGFR) for cell-surface marking of transduced cells. The vector is based on the FMEV backbone which mediates high levels of gene expression in hematopoietic cells. To achieve optimal expression levels of both cDNAs, untranslated regions from MDR1 and DeltaLNGFR were removed and three different connections were tested: retroviral splice signals, an internal ribosomal entry site (IRES) from encephalomyocarditis virus, and an internal promoter from the chicken beta-actin gene. As determined by two-color flow cytometry, the best correlation of the expression of both cDNAs was obtained using the vector SF1mSdelta which utilized retroviral splice signals for co expression. Simultaneous expression of both cDNAs at the single cell level was also shown by confocal laser microscopy. Lymphoid and hematopoietic progenitor cells, including primary human CD34+ cells, transduced with SF1mSdelta acquired dominant multidrug resistance. Transduced primary CD34+ cells could be enriched in vitro based on expression of DeltaLNGFR, avoiding exposure to cytostatic agents. Thus, monitoring the selection of chemotherapy-resistant cells and analyzing their biological properties may be alleviated, both in vitro and in vivo. PMID- 10455431 TI - Recombinant AAV-2 harboring gfp-antisense/ribozyme fusion sequences monitor transduction, gene expression, and show anti-HIV-1 efficacy. AB - Vector-mediated delivery of potentially antivirally active genes is a key step in somatic gene therapy including therapeutic approaches against AIDS. A crucial technical prerequisite is to monitor DNA transfer into target cells. Here, we describe recombinant infectious particles derived from the adeno-associated virus type 2 (AAV-2) that are suitable to deliver effective HIV-1-directed antisense and ribozyme genes into target cells. To monitor transduction, we designed and tested a number of fusions between indicator-coding sequences of luciferase or gfp with effective HIV-1-directed antisense or ribozyme sequences. The combination of an indicator function and an antiviral func- tion in cis allows successful identification of transduced cells and measurement of effects on the replication of HIV-1 in antisense/ribozyme-expressing cells only. The fusion genes were shown to express the indicator genes. Inhibition of HIV-1 replication mediated by the antisense/ribozyme portion of the fusion transcripts was similar to parental constructs and neither acute nor long-term toxicity of fusion genes and their gene products was observed. These results suggest the use of rAAV constructs described here as tools to study the transducibility of target cells, gene expression and efficacy of HIV-1-directed antisense and ribozyme genes. PMID- 10455432 TI - Successful expression of beta-galactosidase and factor IX transgenes in fetal and neonatal sheep after ultrasound-guided percutaneous adenovirus vector administration into the umbilical vein. AB - In utero somatic gene therapy in the later stages of pregnancy may allow targeting of organ systems which are difficult to reach later in life and to prevent the development of tissue damage otherwise caused by the early onset of inherited diseases. We report here on the percutaneous delivery of two adenoviral vectors, containing the beta-galactosidase reporter gene and the human Factor IX gene respectively, to the fetal liver and circulation by ultrasound-guided umbilical vein puncture similar to procedures used in human pregnancy. Vector spread, as detected by PCR analysis for the beta-galactosidase encoding vector, was found in almost all fetal and neonatal organs and in the maternal liver. Expression of the beta-galactosidase transgene was detected in many fetal tissues by RT-PCR. High beta-galactosidase production was shown by immuno-histochemistry predominantly in the liver, where about 30percent of the hepatocytes stained positive, and in the adrenal cortex. Production of factor IX was determined by ELISA in the plasma of treated fetuses and newborn lambs and reached at birth up to 80percent of the normal human plasma concentration. This demonstrates a very hopeful proof of principle for the development of prenatal treatment of many genetic diseases but also requires more detailed investigations with respect to the observed systemic spread of the vector. PMID- 10455434 TI - Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. AB - Development of methods that allow an efficient expression of exogenous genes in animals would provide tools for gene function studies, treatment of diseases and for obtaining gene products. Therefore, we have developed a hydrodynamics-based procedure for expressing transgenes in mice by systemic administration of plasmid DNA. Using cDNA of luciferase and beta-galactosidase as a reporter gene, we demonstrated that an efficient gene transfer and expression can be achieved by a rapid injection of a large volume of DNA solution into animals via the tail vein. Among the organs expressing the transgene, the liver showed the highest level of gene expression. As high as 45 microg of luciferase protein per gram of liver can be achi- eved by a single tail vein injection of 5 microg of plasmid DNA into a mouse. Histochemical analysis using beta-galactosidase gene as a reporter reveals that approximately 40percent of hepatocytes express the transgene. The time response curve shows that the level of transgene expression in the liver reaches the peak level in approximately 8 h after injection and decreases thereafter. The peak level of gene expression can be regained by repeated injection of plasmid DNA. These results suggest that a simple, convenient and efficient method has been developed and which can be used as an effective means for studying gene function, gene regulation and molecular pathophysiology through gene transfer, as well as for expressing proteins in animals. PMID- 10455433 TI - Temporally regulated expression patterns following in utero adenovirus-mediated gene transfer. AB - Developmental patterns of gene expression were determined following intravascular administration of adenovirus in utero, during sequential stages of murine development. Replication-deficient adenovirus (AdCMV.LacZ) was injected into yolk sac vessels of mouse embryos 12, 13, 15 and 18 days post-conception (d.p.c.). beta-Galactosidase (beta-gal) expression was evaluated 24-48 h after injection, at birth, and 5 weeks following normal delivery. Gene expression was detected in myocardial cells, endothelial cells of heart, lung, kidney, adrenal, gut, and in hepatocytes. The patterns of expression were distinct for each stage of virus administration and time-point of analysis. Intensity of individual organ expression varied with injection time-point, with the largest number of organs express- ing the transgene when embryos were injected at 15 d.p.c. beta-Gal activity was detected in only a subset of cells expressing the murine coxsackievirus and adenovirus receptor (CAR), indicating factors other than receptor distribution were responsible for the pattern of transgene expression observed. These studies begin to define critical parameters affecting intravascular gene delivery in utero and indicate that intrinsic developmental regulatory mechanisms may control exogenous gene expression. Intravenous administration of adenovirus provides a unique approach for in utero gene transduction and will be a useful adjunct in evaluating genes which have early lethal mutations. PMID- 10455435 TI - In vivo transduction of mouse epidermis with recombinant retroviral vectors: implications for cutaneous gene therapy. AB - Gene-based therapies may provide a way to treat inherited skin disorders but current approaches suffer serious limitations. The surgical procedures required to transplant ex vivo modified keratinocytes are likely to result in scarring and contracture, thereby limiting the area that can be treated. In addition, none of the methods currently available for in vivo gene transfer to epidermis leads to long-term transgene expression. The goal of this study was to develop a means for in vivo gene transfer to epidermis that would result in long-term transgene expression. We report here the first successful in vivo gene transfer that results in sustained transgene expression in epidermis. Hyperplastic mouse skin was transduced by direct injection of VSV-G pseudotyped retroviral vectors encoding the LacZ reporter gene. In mice tolerant to beta-galactosidase (beta gal), transgene expression was noted in hair follicles and interfollicular epidermis for the duration of the experiment (16 weeks after transduction). Based on the kinetics of epidermal turnover in mouse skin, expression for this length of time strongly suggests stem cell transduction. In immunocompetent mice intolerant to beta-gal, transgene expression was lost by 3 weeks after transduction, concurrent with the onset of host immune responses to the transgene product. PMID- 10455436 TI - A dual specificity promoter system combining cell cycle-regulated and tissue specific transcriptional control. AB - The expression of both proliferation-associated and cell type-specific genes is a hallmark of both cancer cells and tumor endothelial cells. The possibility to combine both features in a single transcriptional control unit would greatly increase the selectivity of vectors used for cancer gene therapy. Previous studies by our laboratory have shown that the transcription of several cell cycle genes is regulated by a novel cell cycle-regulated repressor, termed CDF-1. This repressor functions by blocking in resting cells the transcriptional activation by specific factors binding to the upstream activating sequence (UAS), most notably the CCAAT-box binding factor NF-Y/CBF. Based on this work we have developed a dual specificity promoter system that combines cell type specificity with cell cycle regulation. A chimeric transcription factor (Gal4/NF-Y) consisting of the transactivation domain of NF-Y and the DNA-binding domain of Gal4 is expressed from a tissue-specific promoter. Gal4/NF-Y can bind to a second promoter consisting of a minimal cyclin A promoter with multiple Gal4 binding sites replacing the normal UAS. This leads to the tissue-specific expression of Gal4/NF-Y whose stimulatory activity on the promoter is restrained in resting cells by the recruitment of the CDF-1 repressor to the promoter. The functionality of this system is demonstrated for the specific transcriptional targeting of proliferating melanoma cells, where cell cycle regulation was >20 fold and cell type specificity was >50-fold. PMID- 10455437 TI - Gene targeting is enhanced in human cells overexpressing hRAD51. AB - The ideal therapy for single gene disorders would be repair of the mutated disease genes. Homologous recombination is one of several cellular mechanisms for the repair of DNA damage. Recombination between exogenous DNA and homologous chromosomal loci (gene targeting) can be used to repair an endogenous gene, but the low efficiency of this process is a serious barrier to its therapeutic potential. Recent progress in the isolation and characterisation of mammalian genes and proteins involved in DNA recombination has raised the possibility that the cellular biochemistry of recombination can be manipulated to improve the efficiency of gene targeting. As an initial test of this approach, we have overexpressed the gene encoding hRAD51, a protein with homologous DNA pairing and strand exchange activities, in human cells and measured its effect on gene targeting. We report a two- to three-fold increase in gene targeting, and enhanced resistance to ionising radiation in hRAD51-overexpressing cells with no obvious detrimental effects. These observations provide valuable genetic evidence for the involvement of hRAD51 in both gene targeting and DNA repair in human cells. Our data also establish overexpression of recombination genes as a viable approach to improving gene targeting efficiencies. PMID- 10455438 TI - Mouse adenovirus (MAV-1) expression in primary human endothelial cells and generation of a full-length infectious plasmid. AB - Using RT-PCR, we show that mouse adenovirus type I (MAV-1) is capable of infecting and expressing in various cell types, specifically human endothelial cells. The capability of MAV-1 to infect and express in human endothelial cells makes it a potentially useful alternative to the use of human adenoviruses type 2/5 (Ad2/5) in virus-based gene therapy, although presently MAV-1 can only be produced at lower titers than Ad2/5. In this report, we present methods for the purification of MAV-1 DNA and use of this DNA along with a modified bacteria based homologous recombination protocol to generate a full-length plasmid clone of MAV-1 DNA. Using various transfection procedures, we show that this plasmid MAV-1 DNA can generate plaques of MAV-1 virus, albeit at low efficiencies (about 0. 2 p.f.u./microg DNA). Furthermore, the construction of an MAV-1 plasmid along with its capability to express in human cells justifies the full development of MAV-1 into a system of gene therapy. PMID- 10455439 TI - Adenovirus-mediated gene transfer of the beta2-adrenergic receptor to donor hearts enhances cardiac function. AB - Gene transfer to modify donor heart function during transplantation has significant therapeutic implications. Recent studies by our laboratory in transgenic mice have shown that overexpression of beta2-adrenergic receptors (beta2-ARs) leads to significantly enhanced cardiac function. Thus, we investigated the functional consequences of adenovirus-mediated gene transfer of the human beta2-AR in a rat heterotopic heart transplant model. Donor hearts received 1 ml of solution containing 1 x 1010 p.f.u. of adenovirus encoding the beta2-AR or an empty adenovirus as a control. Five days after transplantation, basal left ventricular (LV) pressure was measured using an isolated, isovolumic heart perfusion apparatus. A subset of hearts was stimulated with the beta2-AR agonist, zinterol. Treatment with the beta2-AR virus resulted in global myocardial gene transfer with a six-fold increase in mean beta-AR density which corresponded to a significant increase in basal contractility (LV + dP/dtmax, control: 3152.1 +/- 286 versus beta2-AR, 6250.6* +/- 432.5 mmHg/s; n = 10, *P < 0.02). beta2-AR overexpressing hearts also had higher contractility after zinterol administration compared with control hearts. Our results indicate that myocardial function of the transplanted heart can be enhanced by the adenovirus mediated delivery of beta2-ARs. Thus, genetic manipulation may offer a novel therapeutic strategy to improve donor heart function in the post- operative setting. PMID- 10455441 TI - Effects of altering dosing on cationic liposome-mediated gene transfer to the respiratory epithelium. AB - Liposome-mediated gene transfer is currently sub-optimal with respect to both the extent and duration of transgene expression. We investigated whether simple changes in DNA dosing could enhance either of these outcomes. Increasing DNA doses produced highest transgene expression at an intermediate dose with toxicity observed at higher doses, thereby likely limiting expression. Adminis- tering an equivalent DNA dose in aliquots over a 1-3 day period resulted in significantly lower gene expression and did not increase the duration of expression. Administration at different times of the day (and hence wake/sleep cycles of the animals) did not alter gene expression. We conclude that such simple changes in dosing regimes are unlikely to contribute to improvements in gene transfer efficiency. PMID- 10455440 TI - Immune response to green fluorescent protein: implications for gene therapy. AB - Green fluorescent protein (GFP) is a widely used intracellular reporter molecule to assess gene transfer and expression. A potential use for GFP is as a co expressed marker, to select and enrich gene-modified cells by flow cytometry. Processed peptides derived from GFP and presented by the major histocompatibility complex on the cell surface could potentially induce T cell immune responses against GFP+ cells. Thus, clinical application of GFP is premature, since in vivo studies on its immunogenicity are lacking. Therefore, we investigated immune responses against EGFP (enhanced-GFP) in two transplantable murine models: the BALB/c (H-2d) BM185 pre-B leukemia and the C57BL/6 (H-2b) EL-4 T cell lymphoma. BM185 and EL-4 cell lines modified to express high levels of EGFP showed drastic reduction of disease development when transplanted into immunocompetent mice. BM185/ EGFP did lead to rapid development of disease in immunodeficient Nu/Nu mice. Mice surviving BM185/EGFP leukemia challenge developed high cytotoxic T lymphocyte (CTL) responses against EGFP-expressing cells. Furthermore, immune stimulation against BM185/EGFP cells could also be induced by immunization with EGFP+ transduced dendritic cells. The effects of the co-expression of EGFP and immunomodulators (CD80 plus GM-CSF) were also investigated as an irradiated leukemia vaccine. EGFP co-expression by the vaccine did not interfere with the development of CTLs against the parental leukemia or with the anti-leukemia response in vivo. These results indicate that the immune response against EGFP may interfere with its applicability in gene insertion/replacement strategies but could potentially be employed for leukemia cell vaccines. PMID- 10455442 TI - Development of human papillomavirus plasmids capable of episomal replication in human cell lines. AB - Gene delivery into human cells has been facilitated by the development of viral vector systems. These vectors have shown great potential for the efficient delivery of therapeutic genes into human cells. A problem with many of the existing systems, however, is the integration of these vectors into the chromosome which affects the length of gene expression and may promote oncogenic transformation. In an effort to develop viral vectors that can replicate extrachromosomally in human cells, we have generated human papillomavirus (HPV) plasmids containing all the elements required for replication on a single DNA molecule. HPV plasmids containing the viral E1 and E2 genes (or the E1 gene alone) and an origin of replication were shown to replicate to significant levels in the transfected human cervical carcinoma C-33A cell line. Since approaches towards the possible gene therapy of cystic fibrosis (CF) are currently under intensive investigation, we have also tested short-term replication of HPV plasmids in the IB3 cell line derived from a CF patient. Our results demonstrate that HPV plasmids are capable of extrachromosomal replication in these cell lines and may potentially be important vectors for the delivery of therapeutic genes into human cells. PMID- 10455443 TI - Titration of AAV-2 particles via a novel capsid ELISA: packaging of genomes can limit production of recombinant AAV-2. AB - We demonstrate the rapid and reliable quantification of physical AAV-2 (adeno associated virus type 2) particles via a novel ELISA based on a monoclonal antibody which selectively recognizes assembled AAV-2 capsids. Titration of a variety of recombinant AAV-2 (rAAV) preparations revealed that at least 80+percent of all particles were empty, compared with a maximum of 50percent in wild-type AAV-2 stocks, indicating that the recombinant genomes were less efficiently encapsidated. This finding was confirmed upon titration of CsCl gradient fractions from recombinant and wild-type AAV-2 stocks. ELISA-based measurement of capsid numbers revealed a large number of physical particles with low densities corresponding to empty capsids in the recombinant, but not in the wild-type AAV-2 preparations. Moreover, additional expression of VP proteins during rAAV production was found to result in an excessive capsid formation, whilst yielding only minor increases in DNA-containing or transducing rAAV particles. We conclude that encapsidation of viral genomes rather than capsid assembly can be limiting for rAAV production, provided that a critical level of VP expression is maintained. The feasibility of quantifying AAV-2 capsid numbers via the ELISA allows determination of physical to DNA-containing or infectious particle ratios. These are important parameters which should help to optimize and standardize the production and application of recombinant AAV-2. PMID- 10455444 TI - Enhanced expression of recombinant dystrophin following intramuscular injection of Epstein-Barr virus (EBV)-based mini-chromosome vectors in mdx mice. AB - Gene transfer by direct intramuscular injection of naked plasmid DNA has been shown to be a safe, simple but relatively inefficient method for gene delivery in vivo. Eukaryotic plasmid expression vectors incorporating the Epstein-Barr virus (EBV) origin of replication (oriP) and EBNA1 gene have been shown to act as autonomous episomally replicating gene transfer vectors which additionally provide nuclear matrix retention functions. Prolonged expression of a LacZ reporter gene and recombinant human dystrophin was shown using EBV-based plasmid vectors transfected into C2C12 mouse myoblast and myotube cultures. Intramuscular injection of EBV-based dystrophin expression plasmids into nude/mdx mice resulted in significant enhancement in the number of muscle fibres expressing recombinant dystrophin compared with a conventional vector. This effect was observed for over 10 weeks after a single administration. These results indicate the potential advantage of EBV-based expression vectors for focal plasmid-mediated gene augmentation therapy in Duchenne muscular dystrophy (DMD) and a range of other gene therapeutic applications. PMID- 10455445 TI - Insertion of an RGD motif into the HI loop of adenovirus fiber protein alters the distribution of transgene expression of the systemically administered vector. AB - Adenoviral vectors are attractive gene delivery vehicles, but their in vivo utility is reduced by lack of cell-specific infection. Tropism modification of the virion by genetic manipulation of capsid proteins is an attractive strategy to achieve targeted transduction. However, no genetic targeting strategies have yet been shown to modify the distribution of transgene expression following systemic administration of vector. This is an essential requirement if such approaches are to form a basis for further vector develop- ment. In this report we present data showing that insertion of a RGD motif into the HI loop of the adenoviral fiber knob results in a significant change in transgene expression profile following intravenous administration. The key finding that a motif in the HI loop is available for cellular interaction when administered systemically means that such modifications can be rationally considered as a foundation upon which further genetic modifications can be superimposed for targeted systemic gene therapy. PMID- 10455447 TI - Ligand substitution of receptor targeted DNA complexes affects gene transfer into hepatoma cells PMID- 10455446 TI - Rapid titer determination using quantitative real-time PCR. AB - Quantitative real-time PCR was utilized to evaluate retroviral vector titer. RNA was prepared from vector supernatant and run in a one-step RT-PCR reaction combining reverse transcription (RT) and amplification in one tube. Sample analysis was performed in the ABI Prism 7700 Sequence Detector. PCR was quantitative over a range of 101 to 6 x 105 vector particles per reaction (2 x 102 to 1 x 107 vector particles per millilites of supernatant) and closely corre- lated with biologic titers performed on the test material. The 96-well capacity of the machine and 2 h of running time permit titer determinations within 8 h, facilitating the processing of large sample numbers while greatly decreasing technician time. Real-time PCR improves titer quantification and the identification of high-titer producer cells. This methodology will help investigators meet the challenges of developing vectors which lack selectable markers. PMID- 10455464 TI - Obesity and type 2 diabetes: a conflict of interests? AB - Obesity is a major risk factor for the development of type 2 diabetes and is an important obstacle to the management of this disease. The increasing incidence of both obesity and type 2 diabetes makes management of these related conditions particularly important. Conventional approaches to the management of type 2 diabetes that focus primarily on improving glycaemic control-notably insulin or sulphonylurea treatment-often lead to weight gain, which is particularly detrimental to patients with type 2 diabetes. By contrast, reducing body weight in such patients improves glycaemic control and other cardiovascular risk factors associated with the metabolic syndrome. This suggests that weight reduction is a rational option in the management of obese patients with type 2 diabetes. While reductions in body weight of approximately 10% have been achieved in some studies, this is difficult to achieve in real life, especially for patients with type 2 diabetes. Weight management agents such as sibutramine and orlistat, used as part of an integrated programme of diet, physical activity and behavioural therapy, are thus an attractive early option for the management of type 2 diabetes in obese patients. PMID- 10455465 TI - The challenge of diet, exercise and lifestyle modification in the management of the obese diabetic patient. AB - Type 2 diabetes is associated with many comorbid medical conditions including obesity, neuropathy, microvascular pathology and atherosclerotic arterial disease. Due to its complications and chronicity, reducing risk factors such as obesity and sedentary lifestyle through lifestyle modification is crucial to the long-term health of patients with type 2 diabetes. Patients must learn how to adopt lifelong, low-fat eating habits and regular activity patterns, with formal treatments focusing on weight loss, increased physical activity and low-fat, low saturated fat diets. In this article we review the efficacy of lifestyle modification programmes for obese diabetic patients. In addition, we discuss barriers to lifestyle changes and methods for improving long-term adherence. Finally, we present information on how this approach has been adapted to a group of Mexican Americans in the USA, a population at high risk for type 2 diabetes, obesity and sedentary lifestyle. PMID- 10455466 TI - The impact of pharmacotherapy on weight management in type 2 diabetes. AB - Today, obesity is the most important modifiable risk factor for type 2 diabetes. An excess of body fat is associated with a deterioration of glucose utilisation and promotes the development of type 2 diabetes, particularly in those with a genetic predisposition for the disease. It is also well established that a reduction of excess body fat improves insulin sensitivity and can prevent the conversion to diabetes. In those with overt diabetes, weight loss usually ameliorates glycaemic control and associated metabolic disturbances. Among the pharmacological agents that are used for the treatment of type 2 diabetes only metformin has a weak weight-lowering activity and is considered as the drug of choice for adjunct pharmacotherapy in obese diabetic subjects. A few studies also suggest that acarbose can induce a modest weight reduction in such patients. In contrast, sulphonylurea and insulin treatment is frequently accompanied by substantial weight gain which should be taken into consideration when these drugs are used. Another approach to improve metabolic control in obese type 2 diabetic patients is the use of weight-lowering agents. The new serotonin and noradrenaline reuptake inhibitor sibutramine promotes weight loss which subsequently leads to improved glycaemic control. Orlistat, a lipase inhibitor, is also able to ameliorate metabolic control in such patients due to its weight lowering potential. As obesity remains a therapeutic challenge in most type 2 diabetic subjects, weight management drugs may represent an alternative or supplement to antidiabetic agents. Moreover, weight management agents have the advantage that they have additional favourable effects on associated cardiovascular risk factors. PMID- 10455467 TI - Patient case studies. AB - Three contrasting cases of obese patients with type 2 diabetes mellitus are presented, which illustrate the management difficulties faced by clinicians. The first raises the issue of when to commence an oral hypoglycaemic agent in a newly diagnosed but asymptomatic obese patient; the second case addresses the problem of when to commence insulin in the face of continuing weight gain and poor glycaemic control; the final case is an example of the vicious metabolic spiral which so many patients enter, with increasing body weight, poor diabetic control and associated co-morbidities. The discussion that follows each case presentation recognises the considerable cardiovascular risk faced by such patients and provides guidance about possible management pathways including adjunctive anti obesity pharmacotherapy. PMID- 10455468 TI - Antihypertensive agents and left ventricular hypertrophy with 'usual' care in the population setting. PMID- 10455469 TI - The influence of different antihypertensive drugs on left ventricular hypertrophy in a population sample of elderly men. AB - AIMS AND METHODS: To evaluate if the effects of long-term treatment with different antihypertensive drugs on left ventricular (LV) hypertrophy seen in meta-analysis could also be found in a cross-sectional study of elderly men, we investigated 584 70-year-old men with echocardiography of which 179 were treated for hypertension. RESULTS: Men on antihypertensive treatment showed a significantly higher LV mass than normotensive men (143 +/- 29 vs 132 +/- 28 g/m2, P < 0.01), but no significant difference in LV mass was seen between men on one antihypertensive drug (n = 129) and those on two drugs (n = 47). Among the men on monotherapy, no differences in LV mass between the subjects who were treated with calcium-antagonists (142 +/- 27 g/m2, n = 37), beta-blockers (140 +/ 29 g/m2, n = 66) or ACE-inhibitors (142 +/- 33 g/m2, n = 20) were found. Neither did blood pressure differ between these groups. Excluding subjects with coronary heart disease did not alter the results. Thiazides as monotherapy was uncommon in this population (n = 4) and therefore not evaluated. CONCLUSION: In a population sample of elderly men with hypertension, no differences in LV mass or blood pressure could be seen between subjects treated with ACE-inhibitors, calcium antagonists or beta-blockers as monotherapy. In meta-analysis of the effects of different antihypertensive therapies on regression of LV hypertrophy, ACE inhibitors have shown the best results. PMID- 10455470 TI - Echocardiographic definition of left ventricular hypertrophy in the hypertensive: which method of indexation of left ventricular mass? AB - OBJECTIVES: It has been suggested that hypertensives at high risk of cardiovascular complications can be identified on the basis of their left ventricular mass as determined echographically. However, there is as yet a lack of consensus on the mode of indexation (body surface area, height, height 2.7) of left ventricular mass (LVM), and on the cut-off values for definition of left ventricular hypertrophy (LVH). The main objective of this study is to test the influence of the different modes of indexation for LVM on the prevalence of LVH in a population of never treated hypertensive patients on the basis of cut-offs for LVM based upon its relationship with ambulatory blood pressure (BP) measurement. METHODS: A population of 363 untreated hypertensives was investigated using a standardised procedure. The men and women were analysed separately. We studied the relationship between mean daytime ambulatory systolic BP and LVM and calculated the LVM cut-off for a BP of 135 mm Hg using three different methods of indexation. On the basis of these criteria, the population was divided into those with and those without LVH. RESULTS: The prevalence of LVH was found to be higher when LVM was indexed to height2.7 (50.4%) or height (50.1%). Prevalence was lowest when LVM was indexed to body surface area (48.2%), which tended to minimise the hypertrophy in obese individuals. Only indexation by height 2.7 fully compensates for relationships between height and ventricular mass in this population. CONCLUSIONS: Indexing LVM to height 2.7 thus appeared to give a more sensitive estimate of LVH by eliminating the influence of growth. Cut offs of 47 g/m2.7 in women and 53 g/m2.7 in men corresponded to a cardiovascular risk indicated by a daytime systolic BP >/=135 mm Hg. PMID- 10455471 TI - The prognostic significance of a history of systemic hypertension in patients randomised to either placebo or ramipril following acute myocardial infarction: evidence from the AIRE study. Acute Infarction Ramipril Efficacy. AB - BACKGROUND: After acute myocardial infarction (AMI), patients with a history of arterial hypertension (AH) have a worse prognosis than normotensives. Whether this adverse risk is beneficially modulated by treatment with angiotensin converting enzyme (ACE) inhibitors is unknown. We evaluated the prognostic value of antecedent hypertension in post-AMI patients given ACE inhibitor therapy. METHODS: We analysed retrospectively data from the AIRE study (randomised, placebo-controlled trial of ramipril in 2006 post-AMI patients with clinical heart failure). A history of AH was present in 28% of the patients. We examined the prognostic value of antecedent hypertension separately among placebo and ramipril treated patients and also the effect of ramipril on clinical outcomes according to whether or not a history of AH was present. RESULTS: Antecedent hypertension was a significant indicator of adverse prognosis in the placebo (P) treated patients (Hazard Ratio 1.49, 95% Confidence Intervals 1.13 to 1.97, P = 0.005) but not in the ramipril (R) treated patients (1.17, 0.84 to 1.61, P = 0.34). A similar pattern was observed for the risks of sudden death (P: 1.75, 1.21 to 2.54, P = 0.003; R: 1.34, 0.86 to 2.07, P = 0.18) and severe/resistant heart failure (P: 1.48, 1.08 to 2.03, P = 0.014; R: 1.18, 0.83 to 1.68, P = 0.37). Treatment with ramipril reduced the all-cause mortality risk in both hypertensive (0.63, 0.44 to 0.89, P = 0.009) and normotensive patients (0.78, 0.61 to 0.99, P = 0.041). CONCLUSION: Antecedent hypertension is not a significant prognosticator in patients with AMI and clinical heart failure given ACE inhibitor therapy. PMID- 10455472 TI - Increased severity of multifocal renal arterial fibromuscular dysplasia in smokers. AB - Renal arterial fibromuscular dysplasia (FMD) is a significant cause of renovascular hypertension, especially in younger females. Tobacco constituents have been shown to stimulate proliferation and synthetic activity of cultured human vascular smooth muscle cells. We examined the relationship between smoking and severity of FMD in a group of 50 subjects with the multifocal form of renal arterial FMD. A detailed smoking history was obtained by interview, clinical data at diagnosis of FMD were obtained from medical records, and angiograms were reviewed. Clinical and angiographic features were compared between smokers and non-smokers. Twenty-four subjects were smokers. At the time of diagnosis of FMD, smokers were of younger mean age than non-smokers (38.7 years vs 48.9 years, P < 0.01), had a shorter median history of hypertension (1.5 years vs 8.5 years, P < 0.05), and had a higher prevalence of unilateral renal atrophy (67% vs 27%, P < 0.01). The distribution of age at diagnosis of FMD was unimodal in non-smokers and bimodal, with a discrete group of younger subjects, in smokers. We conclude that cigarette smoking is associated with an earlier onset and increased severity of disease in a susceptible subgroup of patients predisposed to multifocal renal arterial FMD. PMID- 10455473 TI - Dundee step test: a simple method of measuring the blood pressure response to exercise. AB - The exercise systolic blood pressure (BP) response provides prognostic information over and above that of resting clinic BP in both normotensive and hypertensive individuals. We have developed a 3-min step test as a method of measuring the exercise systolic BP. Healthy volunteers and patients referred for assessment of hypertension took part in validating this exercise test. We assessed the reproducibility of the exercise systolic BP response, and this was compared with that obtained using the cycle ergometry at an equivalent workload. We also compared the baseline characteristics, BP profiles and exercise systolic BP responses in different subject groups. The intra-observer coefficient of variation assessed in 25 subjects was 5.9% with a mean difference of 1.8 mm Hg. The values for between observer were 8.3% and 2.5 mm Hg respectively. Exercise systolic BP measured with the step test correlated with that of cycle ergometer (n = 37, r = 0.93, P < 0.01). Exercise systolic BP data from healthy volunteers (n = 107) showed a normal distribution. An exercise systolic BP of >/=180 mm Hg was greater than 2 standard deviations from the mean and was taken as an abnormally high BP during exercise. There was a positive correlation between exercise BP and increasing age in healthy volunteers (r = 0.57, P < 0.01). This was also present in hypertensive subjects (n = 46, r = 0.48, P = 0.001), however the slope of this relationship was twice as steep as in the normal subjects. Hypertensive subjects with high exercise systolic BP had significantly higher clinic BP, ABPM and a greater BP rise during exercise. The Dundee step test is quick, reproducible and may be prognostically useful. PMID- 10455474 TI - The effects of exercise intensity on post exercise hypotension. AB - Ten normotensive, recreationally active participants aged 35.0 +/- 16.3 years, volunteered to participate in the study. Average baseline blood pressure (BP) was 132/75 mm Hg for systolic (SBP) and diastolic (DBP) pressure respectively. On two separate days, participants underwent testing in a randomised, repeated measures fashion such that they performed 30-min bouts of cycle ergometry at a power output which elicited 50 or 75% of VO2 Peak. Blood pressure was monitored continuously throughout the session by the Finapres method with 2-min windows recorded at rest, 5, 10, 15, 30, 45 and 60 min post exercise. SBP was similar between the two trials and became hypotensive at 5 through 15 min post exercise. The largest decrement (8 mm Hg) in SBP occurred 5 min post exercise. DBP was also unaffected by the intensity of exercise and was lower than before exercise at 5 and 15 through 45 min post exercise. Similarly, mean arterial pressure (MAP) showed significant decrements at 5 and 15 through 45 min post exercise irrespective of exercise intensity. Heart rate was greater during the 75% intensity than during the 50% intensity trial. Pre-exercise values were re established by 45 min post exercise. VO2 remained significantly elevated above pre-exercise values in both trials until 15 min post exercise. Haematocrit increased significantly during both exercise bouts but returned to pre-exercise values by 10 min post exercise. This study indicates that cycle ergometry at 50 and 75% of VO2 Peak elicit similar reductions in post exercise BP. Therefore bouts of mild to moderate intensity exercise may be beneficial in the control of hypertension. PMID- 10455475 TI - Hypertriglyceridaemia in mild hypertension: impact on cardiovascular and hormonal reactivity under different stress tests. AB - OBJECTIVES: We investigated the significance of hypertriglyceridaemia (HTG) for associated components of the metabolic syndrome and stress reactivity in subjects with mild hypertension. METHODS: Seventeen asymptomatic subjects with mild hypertension assessed by 24-h blood pressure (BP) measurement plus HTG (TG >2.3 mmol/l) were recruited and compared with age- and sex-matched subjects with hypertension alone and healthy controls. Cardiovascular and hormonal stress reactivity were tested in a standardised programme consisting of 6 min mental stress, 3 min finger grip and a submaximal bicycle ergometry. RESULTS: The hypertensive patients with HTG exhibited significantly higher fasting insulin, uric acid and gamma-GT levels and lower HDL-cholesterol. The cardiovascular reactivity was similar in all three tests with respect to brachial and peripheral BP in the groups. Peripheral BP during the tests was found to be higher particularly in subjects with hypertension alone. The correlations between BP parameters were disturbed in hypertensives with HTG. Latter group showed significantly higher dopamine, noradrenaline, as well as ACTH levels and an increased ACTH/cortisol ratio. CONCLUSION: HTG in mild hypertension is indicative for insulin resistance accompanied by a modified vascular reactivity as well as elevated catecholamines and ACTH. PMID- 10455476 TI - White coat hypertension and pregnancy outcome. AB - The presence and outcome effect of white coat hypertension in pregnancy was determined with 24-h ambulatory blood pressure (BP) monitoring. Sixty women presenting with high clinic BP (>/=140/90 mm Hg) in the second trimester were included. Patients were divided into two groups based on daytime ambulatory BP findings: <135/85 mm Hg, white coat hypertension (n = 37); >/=135/85 mm Hg, 'true' hypertension (n = 23). Complicated pregnancy outcome was defined as the presence of pre-eclampsia and/or intrauterine growth restriction. Groups were compared for pregnancy outcome and for background and delivery factors. The predictive value of ambulatory BP measurements for pregnancy outcome was determined. Pregnancy outcome was better in the white coat hypertension group than in the true hypertension group: pre-eclampsia-3 (8.1%) vs 13 (56.5%) (P = 0.0046); intrauterine growth restriction-5 (13.5%) vs 10 (43.4%) (P = 0. 0139); and preterm delivery-11 (29.7%) vs 15 (65.2%) (P = 0.015). Night-time ambulatory BP measurements were the best predictor of complicated pregnancy, followed by daytime and 24-h measurements. We conclude that second trimester ambulatory BP monitoring can be used to differentiate patients who have white coat hypertension, which is associated with a better pregnancy outcome than true hypertension. PMID- 10455477 TI - Prevalence of hypertension in a Bangladeshi adult population. PMID- 10455478 TI - Losartan: a study of pharmacovigilance data on 14,522 patients. AB - The objective of this study was to undertake a post- marketing safety surveillance study of losartan, the first specific angiotensin-II receptor antagonist to be marketed in England. It is a non-interventional observational cohort study using the technique of prescription-event monitoring. Subjects who took part in the study were from 14 522 patients treated by general practitioners in England. We recorded adverse drug reactions, adverse events, reasons for stopping the drug, pregnancies and deaths. The results were that general practitioners considered the drug to have been effective in 85.9% of the patients evaluated for effectiveness. Dizziness, malaise, lassitude, nausea, cough and headache were among the most frequently reported adverse drug reactions, reasons for stopping the drug and events of highest incidence density. None of the deaths that occurred were attributed to losartan and no serious unexpected adverse reactions or interactions were identified. In conclusion losartan has an acceptable safety profile and the study confirms the safety information given in the current Summary of Product characteristics for losartan. PMID- 10455479 TI - Impact of antihypertensive treatment on quality of life: comparison between bisoprolol and bendrofluazide. AB - OBJECTIVES: To compare quality of life with the selective beta1-blocker bisoprolol and the thiazide diuretic bendrofluazide in patients with mild to moderate hypertension. DESIGN AND SETTING: Multi centric, randomised, double blind, two-way crossover study carried out at six general practice centres. SUBJECTS: Eighty-one patients with newly diagnosed or previously treated hypertension, who had a mean diastolic blood pressure (BP) of 95-120 mm Hg after receiving placebo for 4-6 weeks. INTERVENTIONS: In random order, patients received bisoprolol (5 mg once daily) or bendrofluazide (2. 5 mg once daily) for 8 weeks. MAIN OUTCOME MEASURES: Quality of life and antihypertensive effect. RESULTS: Decrease in systolic/diastolic BP did not differ between bisoprolol (10 +/- 2/13 +/- 1 mm Hg) and bendrofluazide (9 +/- 2/11 +/- 1 mm Hg). Between bisoprolol and bendrofluazide neither in the intention-to-treat nor in the efficacy analysis any difference was found in quality of life variables, such as Health Status Index, somatic symptoms, anxiety, depression, total psychiatric morbidity, cognitive symptoms and hostility score. Compared to baseline the Health Status Index improved (P < 0.05) during bisoprolol. None of the other investigated quality of life variables changed compared to baseline. No patients dropped out during bisoprolol or bendrofluazide treatment. Although, the total number of reported adverse events appeared lower during bendrofluazide than during bisoprolol treatment, it is unclear whether drug related adverse events also differ between the two drugs. CONCLUSIONS: At equipotent antihypertensive dosages, the effect of an 8-week treatment on quality of life does not differ between the selective beta1-blocker bisoprolol and the thiazide diuretic bendrofluazide. PMID- 10455480 TI - Achilles tendonitis: an unusual complication of amlodipine therapy. PMID- 10455481 TI - Comments on Dr Alderman's letter on my recent editorial. Urinary sodium. PMID- 10455482 TI - Antibiotic MIC/MBC analysis of Bacillus-based commercial insecticides: use of bioreduction and DNA-based assays. AB - Minimum inhibitory concentration (MIC) assays, monitored by colony counts, growth (turbidity) and bioreduction of non-toxic XTT [2,3-bis (2-methoxy-4-nitro-5 sulphophenyl)-2H-tetrazolium-5-carboxanilide, inner salt], were used to assess the performance of several types of antibiotics against: (1) commercial BT products made from scale-up sporulation phase cultures of Bacillus thuringiensis subsp israelensis (Bti) and subsp kurstaki (Btk); (2) vegetative cells derived from these BT products; and (3) Gram-positive and Gram-negative bacteria used as controls. The XTT-kinetic assay improved sensitivity and early reading of MIC breakpoints. The conventional colony count method for determining minimal bactericidal concentration (MBC) was used to validate a multi-sample dot-blot assay in which organisms in individual MIC assays are trapped free of residual antibiotic and their viability is estimated by in situ conversion of MTT [3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] to insoluble formazan. Tolerance (MBC/MIC) for most antibiotics was low ( xylose > arabinose from mixed substrates. Glucose (10 g L-1), ethanol (7.5 g L-1) and acetate (5 g L-1) inhibited xylitol production by 61, 84 and 68%, respectively. Arabinose (10 g L-1) had no inhibitory effect on xylitol production. PMID- 10455494 TI - In vitro and in vivo antagonism of pathogenic turfgrass fungi by Streptomyces hygroscopicus strains YCED9 and WYE53. AB - Disease prevention is a current practice used to minimize fungal diseases of turfgrasses in lawns and golf greens. Prevention is accomplished through fungicide applications, and by periodic thatch removal. During the development of a microbial biodethatch product utilizing the lignocellulose-degrading Streptomyces hygroscopicus strains YCED9 and WYE53, we demonstrated using in vitro plate antagonism bioassays that both strains are antagonists of various turfgrass fungal pathogens. These activities were present when the cultures were growing on thatch, as demonstrated by antifungal antagonism bioassays with culture filtrates. Experiments conducted using a growth chamber demonstrated that a bio-dethatch formulation containing spores of strains YCED9 and WYE53 in a zeolite carrier, provided protection for Kentucky bluegrass seedlings against turfgrass pathogens, including Pythium ultimum, Fusarium oxysporum, Rhizoctonia solani, Sclerotinia homeocarpa, Gaeumannomyces graminis and Microdochium nivale. Results showed that by integrating the use of the S. hygroscopicus YCED9/WYE53 bio-dethatch formulation into routine turf management practices, it should be possible to both minimize thatch build-up while also controlling fungal turfgrass diseases by way of the antifungal biocontrol activity of these strains. This in turn would help control fungal pathogens in turfgrass while minimizing the need for routine chemical fungicide applications. PMID- 10455495 TI - Genetic engineering of doxorubicin production in Streptomyces peucetius: a review. AB - The genetics and biochemistry of daunorubicin and doxorubicin production by Streptomyces peucetius is reviewed, with a focus on how such information can be used for the genetic engineering of strains having improved titers of these two antitumor antibiotics. PMID- 10455496 TI - Detection of bacterial cytotoxic activities from water-damaged ceiling tile material following incubation on blood agar. AB - Samples of ceiling tiles with high levels of bacteria exhibited cytotoxic activities on a HEP-2 tissue culture assay. Ceiling tiles containing low levels of bacterial colonization did not show cytotoxic activities on the HEP-2 tissue culture assay. Using a spread plate procedure on blood agar plates, the levels of bacteria colonizing portions of cellulosic indoor ceiling tiles were easily identified. Levels of bacteria measured by this simple procedure may be a good indicator of microbial colonization of indoor building materials especially in the case of water damage. We suggest that bacterial levels above 150 CFU g(-1) of ceiling tile material indicate colonization has occurred. PMID- 10455497 TI - Some structural features of an insoluble alpha-D-glucan from a mutant strain of Leuconostoc mesenteroides NRRL B-1355. AB - Leuconostoc mesenteroides strain NRRL B-1355 produces two soluble extracellular alpha-D-glucans from sucrose: alternan and dextran. An unusual mutant strain derived from NRRL B-1355 has recently been isolated which produces practically no soluble polysaccharide, but significant amounts of an insoluble D-glucan. Methylation analysis shows it contains linear (1-->3) and (1-->6) linkages as well as (1-->2) and (1-->3) branch linkages. The insoluble glucan was partially digestible by endodextranase, giving rise to a series of oligosaccharides, a high molecular weight soluble fraction and an insoluble residue. Treatment of the soluble dextranase-limit fraction with an alpha(1-->2) debranching enzyme led to further dextranase susceptibility. Methylation, FTIR and NMR analyses of the dextranase-treated fractions indicate a non-uniform structure with domains bearing similarities to L. mesenteroides strain NRRL B-1299 dextran and to insoluble streptococcal D-glucans. PMID- 10455498 TI - Production, purification and characterization of a constitutive intracellular alpha-galactosidase from the thermophilic fungus Humicola sp. AB - The thermophilic fungus Humicola sp constitutively produces intracellular alpha galactosidase (1.33 U mg-1 protein) within 48 h at 45 degrees C in shaken flasks, when grown in a medium containing 7% wheat bran extract as a carbon source and 0.5% yeast extract as a nitrogen source. The enzyme has been purified to homogeneity by ultrafiltration, ethanol precipitation, DEAE cellulose and Sephacryl S-300 chromatography with a 124-fold increase in specific activity and 29.5% recovery. The molecular weight of the enzyme is 371.5 kDa by gel filtration on Sephacryl S-300 and 87.1 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme has an optimum temperature of 65 degrees C and an optimum pH of 5.0. Humicola alpha-galactosidase is a glycoprotein with 8.3% carbohydrate content and is acidic in nature with a pI of 4.0. The KmS for p-nitrophenyl-alpha-D galactopyranoside, O-nitrophenyl-alpha-D-galactopyranoside, raffinose and stachyose are 0.279, 0.40, 1.45 and 1.42 mM respectively. The enzyme activity was strongly inhibited by Ag+ and Hg2+. D-Galactose inhibited alpha-galactosidase competitively and the inhibition constant (Ki) for galactose was 11 mM. PMID- 10455499 TI - A single-use luciferase-based mercury biosensor using Escherichia coli HB101 immobilized in a latex copolymer film. AB - A single-use Hg(II) patch biosensor has been developed consisting of 1.25-cm diameter patches of two acrylic vinyl acetate copolymer layers coated on polyester. The top layer copolymer was 47 microm thick whereas the bottom layer of copolymer plus E. coli cells was 30 microm thick. The immobilized E. coli HB101 cells harbored a mer-lux plasmid construct and produced a detectable light signal when exposed to Hg(II). The immobilized-cell Hg(II) biosensor had a sensitivity similar to that of suspended cells but a significantly larger detection range. The levels of mercury detected by the patches ranged from 0.1 nM to 10 000 nM HgCl2 in pyruvate buffer, and luciferase induction as a function of Hg(II) concentration was sigmoidal. Luciferase activity was detected in immobilized cells for more than 78 h after exposure of the cells to HgCl2. Addition of 1 mM D-cysteine to the pyruvate buffer increased luciferase induction more than 100-fold in the immobilized cell patches and 3.5-fold in a comparable suspension culture. The copolymer patches with immobilized cells were stable at 20 degrees C for at least 3 months, and the Hg(II)-induced luciferase activity after storage was similar to that of samples assayed immediately after coating. Patches stored desiccated at room temperature for 2 weeks showed lower mercury induced luciferase activity when compared to freshly prepared patches, but they still had a considerable detection range of 1 to 10 000 nM HgCl2. PMID- 10455500 TI - Separation of strains of the yeasts Xanthophyllomyces dendrorhousand Phaffia rhodozyma based on rDNA IGS and ITS sequence analysis. AB - The type strains of the anamorph Phaffia rhodozyma (CBS 5905) and the teleomorph Xanthophyllomyces dendrorhous (VKM Y-2786) were analyzed by nucleotide sequence analysis and compared to the sequences found in three additional strains (ATCC 24228, ATCC 24230 and CBS 6938). The results of ribosomal DNA Internal transcribed spacer (ITS) and Intergenic spacer (IGS) region analyses indicate that P. rhodozyma, which was isolated from a beech tree, is a distinct species from the other four strains. The latter that were collected from birch trees are considered to be strains of X. dendrorhous. These individual strains of X. dendrorhous, which have geographically distinct isolation sources, can be distinguished by nucleotide substitutions and deletion/insertion gaps in sub repeat regions of the Intergenic spacer. The conclusions demonstrate that differences in the IGS region provide molecular markers for denoting strains that may differ in their biochemical and physiological capabilities. The hypothesis is presented that strain differences in the IGS region may be useful to demonstrate geographic and host specificity. PMID- 10455501 TI - A new xylanase from a Trichoderma harzianum strain. AB - A new xylanase (XYL2) was purified from solid-state cultures of Trichoderma harzianum strain C by ultrafiltration and gel filtration. SDS-PAGE of the xylanase showed an apparent homogeneity and molecular weight of 18 kDa. It had the highest activity at pH 5.0 and 45 degrees C and was stable at 50 degrees C and pH 5.0 up to 4 h xylanase. XYL2 had a low Km with insoluble oat spelt xylan as substrate. Compared to the amino acid composition of xylanases from Trichoderma spp, xylanase XYL2 presented a high content of glutamate/glutamine, phenylalanine and cysteine, and a low content of serine. Xylanase XYL2 improved the delignification and selectivity of unbleached hardwood kraft pulp. PMID- 10455502 TI - Microbial decolorization of azo dyes by Proteus mirabilis. AB - A bacterium identified as Proteus mirabilis was isolated from acclimated sludge from a dyeing wastewater treatment plant. This strain rapidly decolorized a deep red azo dye solution (RED RBN). Features of the decolorizing process related to biodegradation and biosorption were also studied. Although P. mirabilis displayed good growth in shake culture, color removal was best in anoxic static cultures. For color removal, the optimal pH and temperature were 6.5-7.5 and 30-35 degrees C, respectively. The organism exhibited a remarkable color removal capability, even at a high concentration of azo dye. More than 95% of azo dye was reduced within 20 h at a dye concentration of 1.0 g L-1. Decolorization appears to proceed primarily by enzymatic reduction associated with a minor portion, 13-17%, of biosorption to inactivated microbial cells. PMID- 10455503 TI - Kinetic models for astaxanthin production by high cell density mixotrophic culture of the microalga Haematococcus pluvialis. AB - High cell density cultivation of Haematococcus pluvialis for astaxanthin production was carried out in batch and fed-batch modes in 3.7-L bioreactors with stepwise increased light intensity control mode. A high cell density of 2.65 g x L(-1) (batch culture) or 2.74 g x L(-1) (fed-batch culture) was obtained, and total astaxanthin production in the fed-batch culture (64.36 mg x L(-1)) was about 20.5% higher than in the batch culture (53.43 mg x L(-1)). An unstructured kinetic model to describe the microalga culture system including cell growth, astaxanthin formation, as well as sodium acetate consumption was proposed. Good agreement was found between the model predictions and experimental data. The models demonstrated that the optimal light intensity for mixotrophic growth of H. pluvialis in batch or fed-batch cultures in a 3.7-L bioreactor was 90-360 micromol x m(-2) x s(-1), and that the stepwise increased light intensity mode could be replaced by a constant light intensity mode. PMID- 10455504 TI - Influence of carbon and nitrogen sources on Flavobacteriumgrowth and zeaxanthin biosynthesis. AB - Growth and production of zeaxanthin by Flavobacterium sp were studied using different carbon and nitrogen sources in a chemically defined medium. The best growth was supported by sucrose, but glucose yielded similar carotenoid concentrations. Both asparagine and glutamine stimulated growth and pigment formation. Carotenoid production and glucose consumption increased as a function of asparagine concentration. In the presence of asparagine, high glucose concentrations decreased pigment production without affecting biomass formation. In the absence of glucose, asparagine could not support growth and zeaxanthin production. When compared to the effect of 55 mM glucose, 10 mM oxaloacetate increased growth and carotenoid production. Pyruvate and other intermediates of the citric acid cycle showed a similar stimulatory effect. The intermediates of glycolysis: glucose 6-phosphate and fructose 1,6-diphosphate did not support growth. These results suggest that Flavobacterium sp utilizes asparagine primarily as a nitrogen source for growth and production of zeaxanthin. PMID- 10455505 TI - Comparative stability of ethanol production by Escherichia coli KO11 in batch and chemostat culture. AB - Differing claims regarding the stability of the recombinant ethanologen E. coli KO11 are addressed here in batch and chemostat culture. In repeat batch culture, the organism was stable on glucose, mannose, xylose and galactose for at least three serial transfers, even in the absence of a selective antibiotic. Chemostat cultures on glucose were remarkably stable, but on mannose, xylose and a xylose/glucose mixture, they progressively lost their hyperethanologenicity. On xylose, the loss was irreversible, indicating genetic instability. The loss of hyperethanologenicity was accompanied by the production of high concentrations of acetic acid and by increasing biomass yields, suggesting that the higher ATP yield associated with acetate production may foster the growth of acetate producing revertant strains. Plate counts on high chloramphenicol-containing medium, whether directly, or following preliminary growth on non-selective medium, were not a reliable indicator of high ethanologenicity during chemostat culture. In batch culture, the organism appeared to retain its promise for ethanol production from lignocellulosics and concerns that antibiotics may need to be included in all media appear unfounded. PMID- 10455506 TI - Extracellular deoxyribonuclease production by a thermophile, Streptomyces thermonitrificans. AB - A thermophilic bacterial strain, Streptomyces thermonitrificans, produced high levels of extracellular deoxyribonuclease (DNase) when grown on NBG medium (containing 1% peptone, 0.3% beef extract, 1% glucose and 0.5% NaCl). Maximum DNase activity (140 U x ml(-1)) was obtained, in 24 h, when the culture was grown on modified NBG medium (containing 1.3% beef extract, 1% glucose, 0.5% NaCl and 50 microM Mn2+ at 45 degrees C. The crude enzyme showed higher activity on native DNA than on sonicated and heat denatured DNA. Moreover, addition of Mn2+ in the assay mixture resulted in a significant stimulation (10-15 fold) of the enzyme activity. PMID- 10455507 TI - Adherence of Pseudomonas aeruginosa to inanimate polymers including biomaterials. AB - Cells of Pseudomonas aeruginosa were adhered to polymethyl methacrylate, polyvinyl acetate, polyvinyl chloride, polyhydroxyethyl methacrylate, mixed acrylic, silicone, and natural latex materials. Planktonic bacteria and bacteria that adhered to the test materials were compared for their uptake of either L [3,4,5-3H] leucine or [methyl-3H] thymidine during growth in a minimal medium. Leucine incorporation was reduced and thymidine uptake was negligible in adherent bacteria for up to 8 h following primary attachment by which time cells in the planktonic state showed active uptake of both substrates. These reduced uptake periods correlated with lag phases of growth of adherent cells as determined with a sonication-release plate count procedure and analyses of adenosine triphosphate (ATP). The extent of the lag phase of the adherent populations was dependent on initial densities of adhered cells and the nature of the substratum. PMID- 10455509 TI - Gender, biology and human disease: report of a conference. PMID- 10455510 TI - The epidemiology of gender-discrepant illness. PMID- 10455511 TI - Sex differences in the epidemiology and outcomes of heart disease: population based trends. AB - We examined trends in heart disease (HD) mortality and the delivery of cardiac in Olmsted County, MN. Between 1979 and 1994, women experienced 51% of the total number of HD (ICD9 codes 390-398,402,404-429) deaths (3095). Age-adjusted HD mortality rate declined from 123 per 100,000 (95%CI 102, 144) in 1979 to 81 (67,95) in 1994. The risk ratio (RR) of HD death in 1994 compared to 1979 was 0.69 for women vs 0.53 for men (P = 0.06). This equates to a decline in HD mortality of 2.5%/y in women and 4.2%/y in men. The decline in HD mortality was less pronounced in older age groups (P < 0.001), reflecting a shift of the burden of HD towards women and the elderly. Compared to men, there was less use of stress tests among women, of cardiology visits after stress testing, and of cardiac procedures among women presenting to the emergency room with unstable angina. Further studies are needed to examine causal links between these trends. PMID- 10455512 TI - Loss of the female advantage with cardiovascular disease for women with diabetes. AB - BACKGROUND: The relative risk of cardiovascular disease (CVD) associated with diabetes is greater for women than men, and diabetic women did not experience temporal declines in CVD mortality observed for the general population and diabetic men. OBJECTIVE: To examine sex differences in CVD risk factors for persons with diabetes over time. DESIGN: Population-based historical cohort study. METHODS: The provider-linked medical records of all Rochester, MN, residents assigned a clinical diagnosis of diabetes were reviewed to confirm case status and assign diagnosis date. Data on fasting glucose, obesity, persistent proteinuria, smoking, hypertension, and dyslipidemia were obtained at diagnosis for confirmed incidence cases. RESULTS: There were 1330 diabetes cases 1970- 1989. Compared to men, women at diagnosis were older and more likely hypertensive, had similar levels of fasting glucose and persistent proteinuria, and less likely to smoke. Among persons diagnosed at younger ages, women were more likely than men to be obese. Comparison with published data for the Rochester population revealed the excess obesity and hypertension associated with diabetes were highest for women < age 55 y. Temporal trends in CVD risk factors did not differ between diabetic men and women. PMID- 10455513 TI - Gender differences: the perspective from biology. AB - The remarkable thing about sexual differentiation is its diversity. That males are the heterogametic sex, larger than females, more aggressive than females, and the 'non-default' mode of sexual differentiation are concepts not valid throughout most of the animal kingdom. Sex chromosomes are characteristic only of land animals. In birds, the heterogametic sex is female and the sex chromosomes are not related to those of mammals. External factors such as temperature determine sex in lower vertebrates, and there is no similarity among sex determining genes of different species. The somatic origin of the sex-determining genes and sex chromosomes forces us to ask: what are the other functions of these genes? Because of the obviousness of the sex chromosomes and hormones we may have focused too little on the somatic effects of sex. PMID- 10455514 TI - Sex-based differences in gene transmission and gene expression. AB - The higher prevalence of certain diseases among women suggests involvement of genetic mechanisms linked to the sex chromosomes or of sex-limited gene expression that may be developmentally or hormonally regulated. Analysis of genetic markers and gene expression patterns provides the means for testing hypotheses related to these mechanisms. PMID- 10455515 TI - Microchimerism: implications for autoimmune disease. AB - Recent studies indicate cells traffic between the fetus and mother during normal human pregnancy and that fetal cells persist in maternal peripheral blood for decades after childbirth. The long-term persistence of fetal cells, when considered together with clinical similarities of chronic graft-vs-host disease and autoimmune disease and the female predilection to autoimmunity, led to the hypothesis that microchimerism is involved in some autoimmune diseases. Sources of engraftment applicable to men and to women who have never been pregnant include from a blood transfusion, a twin sibling, or mother. Initial studies lend support to the hypothesis. A role in disease pathogenesis has not been demonstrated, however, and microchimerism is also common in healthy normals. If microchimerism is involved in the pathogenesis of some autoimmune diseases further understanding potentially may lead to new therapeutic strategies. PMID- 10455516 TI - Gender differences in cellular response. AB - Autoimmune and other diseases frequently manifest a sexual discordance that cannot be explained by hormonal differences. Although there is evidence for gender skewing being caused by gonadal hormones, such skewing does not explain an increase of frequency rather than of severity of such diseases in women. To examine if skewing might have a genetic explanation, we have asked how male and female cells devoid of hormonal differences respond to challenges. We used mouse embryonic cells from heart, liver and brain and assessed cellular responsiveness by cell survival. We find that female cells in general show more sensitivity to challenges, such as ethanol, hydrogen peroxide, and camptothecin. Our findings indicate that there is a differential behavior to challenges, in male vs female cells, which may be due to differences in the biological make-up of the cells with regard to gender, and provide preliminary information regarding the feasibility of this type of approach. PMID- 10455517 TI - Gender differences in some host defense mechanisms. AB - Sexual dimorphism exists in the immune response. Both humoral and cell-mediated immunity are more active in females than in males, and steroid gonadal hormones may play an important role in regulating this response. We have documented gender differences in several aspects of neutrophil and macrophage functions elicited by lipopolysaccharide (LPS) (endotoxin) treatment and/or acute ethanol intoxication. In LPS-treated female rats, circulating neutrophils and alveolar macrophages are resistant to the deleterious effects of surgery and anesthesia on phagocytosis observed in male rats. The generation of cytokine-induced neutrophil chemoattractant (CINC) by hepatocytes and Kupffer cells of LPS-treated rats, as well as TNF-alpha secretion by Kupffer cells and alveolar macrophages of acutely ethanol intoxicated rats are also gender dependent. The effects of alcohol on the immune response are expressed differently in males and females. In LPS plus ethanol-treated rats gender differences were noted in terms of adhesion molecule (CD11b/c) expression on circulating neutrophils, and cytoskeletal reorganization in blood-recruited neutrophils and Kupffer cells. Nitric oxide (NO) plays an important role in inflammatory processes. We found gender differences in NO production by alveolar macrophages of LPS-treated rats; this difference was abrogated by ethanol treatment. LPS tolerance and ethanol treatment modulate hepatic NO production in rats in a cell- and gender-dependent fashion, which may exert a protective influence against oxidative injury in the female liver. PMID- 10455518 TI - Estradiol prevents and testosterone promotes Fas-dependent apoptosis in CD4+ Th2 cells by altering Bcl 2 expression. AB - Coxsackievirus B3 (CVB3) induces myocarditis in male BALB/c mice. Female mice are resistant to viral myocarditis, except in the third trimester of pregnancy and postpartum. Cardiac damage is mediated by T lymphocytes activated during virus infection. Th1 (interferon-gamma+) cell responses promote cardiac injury, while disease resistance correlates to preferential activation of Th2 (interleukin-4+) cell responses. CVB3-specific Th1 and Th2 cell clones were established, treated with between 0 and 100 ng/ml 17beta estradiol and 4-androsten-17beta-ol-one (testosterone) for two days, 51Cr-labeled and cultured on FasL-transfected 3T3 cells to determine susceptibility to Fas-dependent apoptosis. Testosterone treatment enhanced Th2 cell lysis while estradiol treatment was protective. Staining of Th2 cells for Bcl 2, an anti-apoptotic factor, indicates that Bcl 2 expression increased in these cells with estradiol but decreased with testosterone exposure. Hormone-induced changes in Bcl 2 expression likely explain the selective survival of Th2 cells in females and prevention of viral myocarditis. PMID- 10455519 TI - Mechanical signaling in the development of postmenopausal osteoporosis. AB - Estrogen deficiency results in increased bone turnover and net bone loss in rats as well as humans. The respective roles of bone turnover and mechanical strain in mediating estrogen deficiency-induced cancellous bone loss were investigated in ovariectomized rats. Ovariectomy resulted in increased bone turnover in long bones. However, cancellous bone was preferentially lost in the metaphysis, a site that experiences low strain energy during normal physical activity. No bone loss was observed in the epiphysis, a site experiencing higher strain energy, despite a similar increase in bone turnover. The role of mechanical strain in maintaining bone balance was investigated by altering the strain history. Mechanical strain was increased or decreased in long bones of ovariectomized rats by treadmill exercise or functional unloading, respectively. Increasing mechanical loading reduced bone loss in the metaphysis. In contrast, decreasing weight bearing accentuated bone loss in the metaphysis and resulted in bone loss in the epiphysis. Finally, administration of estrogen to ovariectomized rats reduced bone loss in unloaded limbs and prevented bone loss in the loaded limbs. These results suggest that estrogen alters the mechanosensory (mechanostat) set point for skeletal adaptation, effectively reducing the minimum strain energy levels at which bone is added. Additionally, these studies suggest that physical activity as well as endocrine status play an important role in maintenance of the female skeleton during aging. PMID- 10455520 TI - Osteoporosis: gender differences and similarities. AB - Although osteoporosis has traditionally been considered a disease of women, men also incur substantial bone loss with aging, and elderly men have age-specific hip fracture incidence rates and vertebral fracture prevalence rates that are at least half those in women. Early postmenopausal bone loss (which results in the syndrome of type I osteoporosis) is due to the direct skeletal consequences of estrogen deficiency, manifested by an increase in bone resorption without an adequate increase in bone formation. Recent evidence indicates that even late postmenopausal bone loss (type II or 'smile' osteoporosis) in women may be due to estrogen deficiency. In particular, the late consequences of estrogen deficiency in elderly women result in abnormalities in calcium homeostasis and increases in parathyroid hormone secretion, leading to increased bone resorption and bone loss. The etiology of bone loss in aging men has remained relatively unclear. Recent evidence from a male deficient in estrogen receptor-alpha and in two males with aromatase deficiency indicate that estrogen may play a significant role in bone metabolism in men. Moreover, several large epidemiologic studies have found that bone mineral density correlates better with serum estrogen than testosterone in aging men. Thus estrogen deficiency may lead to bone loss in men. PMID- 10455521 TI - Gender-related differences in vascular smooth muscle cell proliferation: implications for prevention of atherosclerosis. AB - Premenopausal women have a significant reduction in coronary artery disease compared to age-matched males. Little is known about the mechanism underlying this cardioprotective effect of estrogen. Contradictory evidence has been published and our lack of basic understanding of hormone interactions and bioavailability of different estrogens prevents definitive interpretation of these data. We demonstrate gender-specific effects in the proliferation of coronary artery vascular smooth muscle cells obtained from a sexually mature animal model. Vascular smooth muscle cells are an integral component of the atherosclerotic plaque, and inhibition of cell proliferation by estrogen may be one mechanism by which estrogen exerts its cardioprotective effect. Various types of estrogen may also have different mechanistic actions on the vascular system. No differences are demonstrated in overall estradiol binding in vascular smooth muscle cells obtained from male or female animals: however, differences in c-jun, c-fos and TIEG gene expression were gender related. Inhibition of vascular smooth muscle cell proliferation may have important implications in the prevention of atherosclerotic disease and these studies may provide evidence for the cardioprotective effect of estrogen. PMID- 10455522 TI - Gender and nitric oxide-mediated vasodilation in humans. AB - BACKGROUND: We compared forearm vasodilator responses between females and males after administration of compounds that stimulate nitric oxide (NO) release, act as NO donors, or are NO-independent. METHODS AND RESULTS: We studied nine premenopausal females and nine age-matched males. Forearm blood flow (FBF) was measured with plethysmography. Graded doses of the endothelial-dependent vasodilators acetylcholine and bradykinin and the endothelial-independent vasodilator sodium nitroprusside were given via a brachial artery catheter to the non-dominant arm. Reactive hyperemia was measured as an index of NO-independent vasodilation. In females estradiol levels were greater, and serum triglycerides were lower, but cholesterol was similar to that of males. FBF responses to all doses of the three drugs were approximately 30% less in females. Females had smaller forearms and lower peak FBF during reactive hyperemia. Blood flow responses to the drugs as a fraction of the peak reactive hyperemia values were similar in females and males. In the females, but not in the males, FBF responses to all three drugs and reactive hyperemia correlated with estradiol levels. CONCLUSIONS: The smaller FBF responses in females were likely due to smaller forearm volume and muscle mass. Estradiol levels are associated with both NO mediated and non-NO-mediated dilator responses in females. PMID- 10455523 TI - Gender and vascular reactivity. AB - Estrogen receptors are found on vascular endothelial and smooth muscle cells; their expression is influenced by exposure to the hormone. Estrogen receptors influence non-genomic events, which are rapid in onset and genomic events, which are longer acting responses. Estrogens affect vascular tone indirectly by modulating release of endothelium-derived vasoactive factors and directly by modulating intracellular calcium in vascular smooth muscle cells. Estrogens indirectly affect thrombotic events and inflammation by altering platelet aggregation and leukocyte adherence and migration, respectively. Estrogens also influence production of mitogens which, when released at sites of vascular injury, affect vascular remodeling. Although estrogens initiate vascular responses, genomic sex may influence and/or limit expression of estrogen receptors and therefore actions of sex steroid hormones throughout the vasculature. PMID- 10455527 TI - Gait after spinal cord injury and the central pattern generator for locomotion. PMID- 10455528 TI - Anterior dislocation and extruded disc of the lower cervical spine. PMID- 10455529 TI - Kinetic factors determining wheelchair propulsion in marathon racers with paraplegia. AB - OBJECTIVE: To assess the different factors that determine the propulsion of wheelchair in long distance racers with spinal cord injuries. DESIGN AND METHODS: Nine highly handicapped men who competed at the Oita International Wheelchair Marathon in 1995, participated in this study. Subjects performed steady and exhaustive wheelchair propulsion at their optimal speed on wheelchair rollers in a laboratory setting during a 5-min period. Movements of the trunk and arms were filmed by a video camera then analyzed kinetically using a two-dimensional analysis system. In order to determine the actual pushing time and angle of wheelchair, we measured the duration between hand contact and hand release from handrim. SETTING: Department of Rehabilitation Medicine University of Occupational and Environmental Health, Japan. RESULTS: The push time, cycle time, total push time, and angular velocity did not relate with the race time or wheelchair velocity. However, the total push angle correlated significantly (P<0.01) with the race time and wheelchair velocity. Improvement of the pushing angular velocity and pushing duration did not contribute to the race time. CONCLUSION: The skilful wheelchair propulsion in long distance racing depends on the total push angle. Improvement of the total push angle should be achieved by both prolongation of the total push time and increased pushing angular velocity. PMID- 10455531 TI - Characteristics of 'low risk' and 'high risk' dives by young adults: risk reduction in spinal cord injury. AB - STUDY DESIGN: To establish benchmark normative data for dive entries performed by young adults of the age range most likely to sustain a diving spinal cord injury. Data acquired from analysis of the dives performed, along with survey information, were used to determine which factors make the most contribution to the level of risk in diving. OBJECTIVES: To identify influential variables which could contribute to risk of spinal cord injury for each of four types of dives. The types of dives investigated were: dive entries from deck level to tread water (Treadwater); deck level to swim 25 m (Deck); starting block height to swim 25 m (Block); and a running dive entry to swim 25 m (Running). SETTING: Victoria, Australia. METHODS: Ninety-five first year university students (average age 19.9 years) performed three or four dives which were video-recorded for later analysis. Maximum depth reached was used as an indicator of risk, and velocity at maximum depth, distance at maximum depth, angle of entry and flight distance were measured for each dive. Participants also completed a questionnaire designed to elicit information about their swimming and diving background. Unlike previous diving studies, participants were recreational rather than competitive swimmers. They were not aware that the dive was the focal point, assuming that the researchers were investigating their swimming and treadwater ability. RESULTS: A stepwise multiple regression was applied to predict depth for each dive condition, and demonstrated that four variables were able to account for 56% of the variance for Treadwater, 68% for Deck; 73% for Running and 79% for Block. In all conditions involving swimming after the dive (ie Deck, Block and Running), beta weights showed that distance at maximum depth had the greatest influence on the depth of a dive. Flight distance and angle of entry were the next most influential variables. For the Treadwater condition, beta weights showed angle of entry was the most influential variable, followed by velocity at maximum depth, distance at maximum depth and swim rank. CONCLUSION: It is recommended that divers strive to surface in as short a distance as possible by maximising flight distance and aiming for a low entry angle. Implementation of steering-up techniques will assist in minimising dive depth. PMID- 10455530 TI - Contribution of D-Dimer determination in the exclusion of deep venous thrombosis in spinal cord injury patients. AB - Deep vein thrombosis (DVT) is a common complication of paraplegia despite prophylactic anticoagulant therapy. The diagnosis relies primarily on ultrasonography or phlebography; these investigations are difficult, expensive and can be time-consuming in paraplegic patients. STUDY DESIGN: To evaluate the usefulness of coagulation activation markers in excluding a diagnosis of DVT, D Dimers, thrombin-antithrombin complexes, prothrombin fragments (F1+2) and activated factor VIIa. OBJECTIVES: To improve the diagnosis of deep venous thrombosis in paraplegic patients. SETTING: This collaborative work was done at Raymond Poincare Hospital, Garches, France. METHODS: To evaluate the usefulness of coagulation activation markers in excluding a diagnosis of DVT, D-Dimers (D Di), thrombin-antithrombin (TAT) complexes, prothrombin fragments (F1+2) and activated factor VIIa (FVIIa), were determined in a prospective study of 67 consecutive patients with paraplegia or tetraplegia. Doppler ultrasonography and/or phlebography of the lower limbs and D-Di, TAT, F1+2 level determination were systematically done in each patient at admission to our rehabilitation unit. RESULTS: Despite prophylactic low molecular weight heparin therapy, six of the 67 patients developed DVT diagnosed by radiologic explorations. D-Di levels measured by a reference ELISA (Asserachrom D-Di, Diagnostica Stago) or a new rapid automated turbidimetric test (STA-Liatest D-Di) were greater than 500 ng/ml in all DVT patients and in 40 non-DVT patients, of whom most had urinary tract infections, osteomas, or pressure sores. D-Di values were normal in only 21/67 patients (31%). The negative predictive value of D-Di in our study was 100% since all DVT patients had D-Di values greater than 500 ng/ml. TAT and F1+2 levels were not correlated with D-Di levels but also had a negative predictive value of 100%. Comparison of D-Di levels obtained using the two tests showed that results of the reference ELISA were closely correlated to those of the new rapid automated turbidimetric. TAT, F1+2, and factor VIIa are not useful for measuring hypercoagulability in paraplegic or tetraplegic patients since no rapid tests for determining these parameters are available. CONCLUSION: D-Di levels determined using an ELISA or a new rapid automated turbidimetric test have a good negative predictive value for DVT in paraplegic or tetraplegic patients and may reduce the need for Doppler ultrasonography and/or phlebography by 31%. PMID- 10455532 TI - Characteristics of injuries to the cervical spine and spinal cord in polytrauma patient population: experience from a regional trauma unit. AB - STUDY DESIGN: Retrospective analysis of a prospectively collected trauma database of a Level 1 (tertiary) trauma center. OBJECTIVE: To define the features of the cervical spinal injuries in polytrauma population admitted to the regional trauma unit. SETTING: Canada, Ontario Province, Toronto, Sunnybrook Health Sciences Center. METHODS: All trauma admissions between 1987 and 1996 entered prospectively into a trauma registry database were studied for incidence, demographic and epidemiological details of cervical spine (cord and column) injuries. RESULTS: A total of 468 patients (66% male) with cervical spinal injury (CSI) from 1198 spinal injuries admitted to the regional trauma center were identified. Seventy-five per cent of the CSI involved were aged less than 50 years; nearly 30% were in the third decade alone. Overall, the commonest spinal level injured was C2 (27%) followed by C5 (22%). Older population (above 60 years of age) had C1 + 2 involved more often than the young (P=0.02). Motor vehicular crashes (MVC) accounted for 71%, followed by pedestrian trauma (10%), sport injuries (7%). Spinal cord injury (SCI) was noted in 27%; complete in 16% and incomplete in 11% and more frequently at C4 or C5 level compared with C1, C2 (P<0.00001); the former level had more often a complete SCI (P=0.06). Though MVC produced 74% of SCI, only 27% had neurological deficits. Recreational trauma produced SCI in 45%, motor cycle crashes (MCC) in 37% and a rear passenger in MVC in 34% that was complete in 78%, 71% and 73% respectively. Front seat passenger and driver in MVC had a C5 level injury while a rear seat passenger had at C4 (P<0.001). The C1 level injury had high association with severe and life threatening head and neck and facial injuries compared with the more frequently injured spinal levels; either C2 (P=0.03) or C5 (P=0.004). Similarly C1 injuries had higher ISS compared with C2 (P<0.0001) and C5 (P<0.008). CONCLUSIONS: C2 was the commonest fractured spine while SCI was more frequent at C5. Older and pedestrian population had higher incidences of injuries at C1 and C2. Sport and MCC resulted in severe SCI. The level of spine injured was different between a front and a rear seat occupant in MVC. PMID- 10455533 TI - Comparison of asynchronous versus synchronous arm crank ergometry. AB - STUDY DESIGN: A direct comparison of synchronous versus asynchronous arm crank ergometry has not been carried out previously. Therefore, a comparative research design was employed. OBJECTIVE: To assess the physiological responses of arm cranking when performed asynchronously (arms moving opposite to each other) versus synchronously (both arms moving in the same direction simultaneously). SETTING: A university hospital setting in Galveston, Texas, USA. METHODS: Seventeen individuals between the ages of 19 and 53 years were studied, 11 with paraplegia and six with no apparent disability. Two maximal arm crank graded exercise tests were performed with the subject seated in a wheelchair. Testing consisted of both arms (1) asynchronously (reciprocally) pushing and pulling the crank handles and (2) pushing and pulling the crank handles synchronously. Each test consisted of 2 min stages starting at 20 W and increasing 10 W per stage thereafter until exhaustion. Heart rate, oxygen consumption, and minute ventilation were measured and recorded during each stage. Blood lactate levels were monitored before and after each test. Statistical analysis was performed using the multivariate Hotelling's T2 followed by post hoc univariate tests. RESULTS: Greater power and longer test times (both groups, P<0.05) and higher post test blood lactates (nondisabled P<0.01, paraplegic P<0.05) were achieved with asynchronous cranking versus synchronous cranking. While submaximal responses were similar between the two modes of cranking, there was a tendency for all variables to be lower with asynchronous. All subjects preferred asynchronous rather than synchronous cranking. CONCLUSION: Despite few statistically significant differences, based on the subjective reports from all subjects, we believe there is a clinically significant difference between the two modes of cranking. The results suggest that the mode of cranking may have implications for arm crank testing, training, and functional locomotion in individuals with lower extremity impairments. PMID- 10455534 TI - Training of the respiratory muscles in individuals with tetraplegia. AB - STUDY DESIGN: An experimental cross-sectional design. OBJECTIVES: To evaluate whether training of the innervated respiratory muscles in individuals with a (partial) cervical spinal cord injury will improve the strength and endurance capacity of these muscles and the exercise performance in these individuals. SETTING: Department of Physiology and Pulmonary diseases, Nijmegen, The Netherlands. METHOD: In this study nine individuals with tetraplegia (C3 C7) performed a target flow endurance training of the inspiratory muscles, twice a day for 15 min. First, the subjects performed a 'sham' training for 6 weeks with no appreciable resistance, after that they performed a 'real' training for 6 weeks with a resistance of 70% of the maximal endurance capacity of the inspiratory muscles. The training was evaluated at 0, 6 and 12 weeks by the following tests: (1) the slow Inspiratory Vital Capacity (IVC) and the Forced Inspiratory and Expiratory Volumes over 1 s (FIV1 and FEV1); (2) the Maximal Inspiratory Mouth Pressure (Pimax) and the Endurance Pressure (Pendu) and (3) a maximal arm-cranking exercise test. RESULTS: After the sham training, the Pendu was increased from 3.98 to 4.71 kPa with a P-value of 0.05. The sham training had no influence on any of the other variables. The real training had no effect on the IVC, FIV1, FEV1 and Pimax, however, increased the Pendu from 4.71 to 6.16 kPa (P=0.01), representing the respiratory muscle-endurance capacity. The oxygen consumption (VO2 peak) in the maximal exercise test improved from 0.87 to 0.98 l/min (P = 0.05). CONCLUSION: The results of the study indicate that training of the respiratory muscles results in an enhanced endurance capacity of these muscles and a concomitant increase in the aerobic exercise performance. PMID- 10455535 TI - Experimental study on MRI evaluation of the course of cervical spinal cord injury. AB - STUDY DESIGN: An experimental study was conducted to evaluate MRI signal changes within the spinal cord after the injury in rats. OBJECTIVES: To clarify the significance of MRI signal changes in the injured cervical cord. SETTING: Tokyo, Japan. METHODS: Cervical spinal cord injury was produced in rats by placing a 20 g, or 35-g weight on exposed dura at the C6 level for 5 min (20 g- or 35 g compression group). Motor function was evaluated by the inclined-plane method at 2, 7 and 28 days after the injury. T1- and T2-weighted images were produced by the spin-echo method with a static magnetic field strength of 2.0 tesla, at 2 and 28 days after the injury, and then the histopathological examinations were performed. RESULTS: In the 20-g compression group, which recovered from the paralysis at 28 days, MR images were T1 iso signal/T2 high signal 2 days after the injury and T1 iso signal/T2 high signal after 28 days. The changes in MRI signal 2 and 28 days represented edema and gliosis, respectively. In the 35-g compression group, which incompletely recovered from paralysis at 28 days. MR images were T1 iso-signal/T2 low signal surrounded by high signal 2 days after the injury and T1 low/T2 high signal at 28 days. The MRI signal changes 2 and 28 days reflected hemorrhage with edema and cavities, respectively. CONCLUSION: T2 low signal of the spinal cord observed early after injury reflects hemorrhage and may serve as an indicator of a poor prognosis. T1 low/T2 high signals from the subacute to chronic period indicated persistence of paralysis and limited recovery of function. PMID- 10455536 TI - Evaluation of the lower motor neuron integrity of upper extremity muscles in high level spinal cord injury. AB - PURPOSE: To evaluate the lower motor neuron (LMN) integrity of upper extremity muscles of persons with high tetraplegia (C1-C4) in order to determine muscles available for stimulation. METHODS: Fourteen subjects (23 arms) were evaluated for LMN integrity. Muscles that elicited a functional response (grade 3 or better) to surface electrical stimulation were considered to have intact LMN and good candidates for FES. Strength-duration (S-D) curves were generated on muscles that showed weak (less than grade 3) or no response to surface stimulation. Muscles were considered denervated if S-D curves were discontinuous or depicted steep, increasing amplitude for pulse durations greater than 1 m. RESULTS: Muscles for grasp and release had intact LMN in 19 of 23 (83%) arms. The wrist extensors and flexors and pronator were excitable in 17 (74%), 20 (87%) and 19 (83%) arms, respectively. The supinator demonstrated LMN lesion in 80% of the arms. Over 90% of the biceps muscles were unresponsive to electrical stimulation and 85% and 87% of the deltoid and supraspinatus muscles, respectively, were not electrically excitable. The latissimus dorsi and triceps muscles were typically innervated (78% and 91%, respectively) and slightly more than half (52%) of the pectoralis major muscles were excitable. CONCLUSION: These data suggest that application of FES in high tetraplegia for hand and arm function would require augmentation because of the inability to stimulate the elbow flexors, deltoid and rotator cuff muscles. These data also show that several paralyzed proximal muscles with intact LMN that have been historically transferred to address shoulder paralysis in other patient populations are available for transfer and stimulation in the population with high level spinal injuries. PMID- 10455537 TI - Liquorpheresis (CSF filtration) in familial amyotrophic lateral sclerosis. AB - OBJECTIVES: Liquorpheresis (CSF filtration) has been shown to be of benefit in various neurologic disorders, including sporadic ALS. Liquorpheresis in patients with familial ALS, has not been investigated so far. METHODS: A 52-year-old woman with familial ALS is reported who underwent liquorpheresis during 4 consecutive days. During this period, 875 ml CSF were filtered off via an intrathecal catheter and a combined mechanical and ionic filter by means of a bi-directional syringe pump. RESULTS: Immediately after treatment and 2 weeks later there was subjective, but no objective, improvement of her symptoms, assessed by the Norris score and measurements of the elbow extension, vital capacity, F-waves of both ulnar nerves and automatic EMG of the right brachial biceps and anterior tibial muscles. CONCLUSION: Liquorpheresis does not seem to be helpful in the treatment of familial ALS. PMID- 10455538 TI - What is the optimum fluid intake in male patients with spinal cord injury and neuropathic bladder? PMID- 10455553 TI - Artful imaging of the brain. From logo to metaphor, about Michelangelo and Kiefer. AB - Michelangelo has painted the fresco 'Creation of Adam' for the Sistine Chapel. Recently it was discovered that he had an outline of the brain concealed in Gods cloud. As a renaissance artist he was not supposed to believe that the brain is the seat of the soul. Only when the concept of structure and function of the central nervous system had developed, artistic presentation of the brain did become possible. The conceptual painter Kiefer, who is possessed by Germany's recent and past history, expressed his apprehension using the philosopher Heidegger and his writings as symbol and a naked brain as metaphor. PMID- 10455554 TI - Fluoride enhances the effect of aluminium chloride on interconnections between aggregates of hippocampal neurons. AB - The role of fluoride in aluminium neurotoxicity was studied using an in vitro system of cultured hippocampal neurons from foetal rats. Sodium fluoride (50 microM) and aluminium chloride (12.5 microM) were administered alone or in a specific combination (50 + 12.5 microM) in a 14-day culture in a chemically defined medium before staining of neurofilaments. Neuronal aggregates interconnected by neuritic fibers were detected light microscopically in control cultures. The aggregates and the fibers stained positive for neurofilament proteins. In cultures treated with aluminium chloride the development of the interconnecting fibers was affected, resulting in a fusion pattern of the aggregates. This phenomenon was enhanced when sodium fluoride was given together with aluminum chloride. It was concluded that aluminium interferes with the metabolism of the neuronal cytoskeleton and that this interference is potentiated by fluoride. PMID- 10455555 TI - Effects of pentanoate, 4-pentenoate, 3-hydroxybutyrate and insulin on the tissue levels of long-chain acyl CoA and acylcarnitine in the oxygenated and hypoxic rat atria. AB - Under hypoxic conditions the atrial contents of long-chain acyl CoA (LCCoA) and long-chain acylcarnitine display a close correlation with the noxious effects of fasting on the atrial functions as well as with the amelioration effected by inhibitors of carnitine palmitoyltransferase I. These findings suggested that fatty acid oxidation was detrimental for the hypoxic atria. However, since changes of the LCCoA and LCCa levels which may occur together with the hypoxic disturbances attained under some other metabolic interventions had not been assessed yet, present investigation aimed to provide information about this issue. At the end of the prehypoxic equilibration period, all the treatments tested evoked a fall of the free-CoA levels whereas free-carnitine, LCCoA and LCCa remained unchanged. In the hypoxic atria, 4-pentenoate, an inhibitor of fatty acid beta-oxidation that also can be oxidized, did not change LCCoA and LCCa levels whereas the readily oxidizable pentanoate evoked a drop of LCCoA. These effects may be due to the trapping of CoA as the short-chain acyl esters of both substances. Since 4-pentenoate and pentanoate were noxious on the hypoxic atria even though they did not increase LCCoA and LCCa contents, it may be inferred that short-chain acyl esters might be deleterious during oxygen shortage. The exposure to 3-hydroxybutyrate, an oxidizable substrate whose availability increases during fasting, did not alter the LCCoA and LCCa contents, agreeing with the weak detrimental effects that it exerts on the hypoxic atria. On the other hand, insulin elicited a rise in the LCCoA and a fall in the LCCa contents. Inasmuch insulin had been shown to improve the performance of the hypoxic atria, these findings suggest that LCCoA might not be involved in the noxious effects of fatty acid oxidation whereas LCCa would be the major toxic catabolite. PMID- 10455556 TI - Protection against ischemia-reperfusion induced oxidative stress by vitamin E treatment. AB - The rat heart protection offered by vitamin E against oxidative stress after ischaemia-reperfusion was studied by using a new methodological approach. Functional recovery of hearts from ischaemia-reperfusion was correlated with a traditional index of oxidative stress such as lipid peroxidation and with antioxidant capacity and susceptibility to oxidants of the tissue evaluated by enhanced chemiluminescence techniques. Rats were treated with ten daily i.m. injections of 100 mg/kg body weight of vitamin E. The functional recovery during reperfusion (20 min, following 45 min ischaemia) of Langendorff preparations from control (vehicle-injected) and vitamin E treated rats was evaluated in terms of heart rate, left ventricular developed pressure (LVDP), double product (= heart rate. LVDP) and coronary flow recovery. Vitamin E treatment significantly improved functional recovery of heart rate, LVDP, double product and coronary flow. It also increased the level of vitamin E and reduced the levels of both malondialdehyde and hydroperoxides in the heart tissue at the end of the ischaemia-reperfusion protocol. In contrast, it did not affect the antioxidant capacity and the response of heart homogenates to in vitro oxidative stress measured after ischaemia-reperfusion. These results show a protective action of vitamin E treatment against lipid peroxidation and cardiac dysfunction associated with ischaemia-reperfusion. Although the precise mechanism of this protection is not evident, our model in part suggests a role of vitamin E other than as a free radical scavenger. PMID- 10455557 TI - Automatic determination of the transition between successive control mechanisms in upright stance assessed by modelling of the centre of pressure. AB - A recently introduced concept models the trajectory of the centre of pressure as a fractional Brownian motion and reveals that two successive scaling regimes, acting hypothetically as open and closed loop mechanisms, are implicated in posture control. Objectivity is obviously required in the determination of the transition point, i.e. the point at which an open-loop control mechanism would switch to a closed-loop one, in order to provide reproducibility and automatism in the processing of data. In the method proposed herein, the transition point corresponds to the maximal distance separating a diffusion curve in a double logarithmic plot (mean square distances MSD calculated on each axis versus increasing time intervals Deltat) from a straight line characterising a pure stochastic behaviour. In closed eye conditions, the switch appears medio laterally in a 0. 26-0.52 s range for Deltat, the corresponding MSD being in the range of 1.86-10.50 mm(2). In the forward-backward direction, the transition is in a 0.28-0.42 s range and the corresponding MSD is between 3.60 and 15.17 mm(2). Finally, these co-ordinates induce scaling exponents over 0.50 for the shortest Deltat, thus suggesting open-loop control, whereas those of longest Deltat, ranged between 0 and 0.20, give evidence of close-loop control. This data is compared to previous data based upon empirical methods. PMID- 10455558 TI - Two-step purification of cytochrome P-450 from adult pig testis by isoelectric focusing. AB - Adult testicular cytochrome P-450 was purified by a two-step procedure utilizing preparative isoelectrofocusing. Purification was achieved 1132 times with a yield of 4.82%. 17alpha-hydroxylase activity was shown to be 14.5 nmol of product/min/nmol of P-450. The cytochrome P-450 was determined to have an isoelectric point of 6.45 on analytical isoelectric focusing. The purified cytochrome P-450 was found to be homogeneous and its molecular weight was estimated to be 52000 on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The carbon monoxide difference spectrum with a peak at 448 nm exhibited the absorption spectrum of a typical cytochrome P-450. PMID- 10455559 TI - Influence of age and sex in serum osteocalcin levels in thoroughbred horses. AB - In this study, we assessed the potential value of free serum osteocalcin or bone gla protein (BGP), the most abundant non collagenous matrix protein found in bone and dentin, to reflect changes of bone turnover in thoroughbred horses. Levels of osteocalcin were analyzed in serum samples of 54 clinically normal animals divided into three groups (A, B, C) according to age: 8, 16-18 and 24-36 months, in order to determine the standard for young horses of different age and sex. Serum BGP was measured by an in-house developed double antibody radioimmunoassay using bovine antigen. The mean BGP levels (ng/ml) were 45.65 +/- 11.69; 33.65 +/- 16.65; 15.08 +/- 6.70 respectively for groups A, B and C; statistically significant differences were found between groups (A vs B and C; Bvs C). Difference between males and females was found significant in group C with higher values in the females: 18.75 +/- 5.00 against 14.43 +/- 10.47 i n the males. This can be considered a sex related effect on BGP serum levels after the onset of puberty. Correlation coefficient between age and serum BGP for females and males were r 5 20.598 ( P < 0.001) and r 5 200.807 (P < 0.001) respectively. A significant negative linear relationship could be established between these two parameters in males during the growth period. The regression equation between serum BGP and age for males was (month of age = 65.14-1.68. BGP). In the female group the gestation and lactation are variables that lower the correlation coefficient between age and serum BGP levels. These results suggest that serum BGP decreases in thoroughbred horses during the growth period, and significant differences between sexes were found only after the onset of puberty. PMID- 10455560 TI - Communities of caregivers: revisiting the settlement model. PMID- 10455561 TI - Hypertensive update: highlights from the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure. AB - The National Heart, Lung and Blood Institute's Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure (JNC VI) proposes a set of public health challenges to fight hypertension. The report proposes a system of risk stratification and pharmacologic therapy. Ongoing care of the person with high blood pressure (HBP) emphasizes the role of the primary care provider. PMID- 10455562 TI - A five-year-old with a dental abscess: a case study. AB - Dental caries remain one of the most common disorders of childhood in the United States. Often nurse practitioners (NPs) will see children who are suffering from the complications of a dental carie, such as a dental abscess and/or cellulitis. This article describes the case of a 5-year-old girl who presented at an evening clinic with tooth pain, fever, and facial swelling. Three treatment choices are discussed: (1) 400 mg of amoxicillin (Augmentin), by mouth, with comfort measures, and return to the clinic in the morning; (2) 2 g of ceftriaxone by injection, with comfort measures, and return to the clinic in the morning; (3) or hospitalize via emergency department for intravenous fluids and antibiotics. The treatment that was chosen not only takes into account the disease process, but also the impact of this choice on the family. A model for the progression of dental caries in low-income groups with recommendations for prevention is also presented. PMID- 10455563 TI - Bell's palsy in the primary care setting: a case study. AB - Bell's palsy is an acute unilateral paralysis of the facial muscles innervated by the seventh cranial nerve. Although the etiology of Bell's palsy is unknown, it is thought to result from edema, entrapment, or inflammation of the seventh cranial nerve. A history of recent viral infection--especially herpes simplex- diabetes mellitus, pregnancy, or hypertension are all common risk factors that may precede the onset. This article reviews assessment findings, differential diagnoses, and the treatment of Bell's palsy, which is considered a diagnosis of exclusion. A 10-day course of oral corticosteroids is the recommended therapy for lessening its course and severity in some populations. Recent research recommends the addition of acyclovir for 10 days, suggesting a herpetic viral etiology. Close follow-up is imperative to prevent corneal injury and to monitor worsening of symptoms. Although most patients recover within 1 to 6 months, incomplete recovery may be seen in severe or recurrent cases. Indications for referral are discussed. A case study is presented to illustrate the findings in this interesting disease. PMID- 10455564 TI - Bacterial vaginosis: a review. AB - Bacterial vaginosis (BV) is a clinically definable condition that is frequently diagnosed in the primary care setting. Changes in the nomenclature, causative microorganisms, as well as treatment have caused confusion over the last century. BV is currently viewed as a shift in the normal vaginal ecosystem from a predominance of aerobic bacteria to anaerobic bacteria (Soper, 1993), resulting in a malodorous discharge. However, more than one half of clinically diagnosed patients are asymptomatic. Recent evidence indicates that the anaerobes identified as components of BV have been implicated in obstetric complications, postoperative pelvic infections, and pelvic inflammatory disease. Practice issues resulting from this research include the appropriate treatment of asymptomatic women, especially in pregnancy. This article reviews BV from a historical perspective, provides a summary of recent research findings identifying BV as a possible risk factor in various serious medical conditions and their sequelae, and discusses related practice concerns resulting from the implications of these findings. PMID- 10455565 TI - Cervical intraepithelial neoplasia diagnosis: emotional impact and nursing implications. AB - Cervical intraepithelial neoplasia (CIN) is a progressive disease process that starts with abnormal cellular development in the cervical epithelium and can be identified as low- or high-grade squamous intraepithelial lesions (LGSIL or HGSIL) by the Papanicolaou (Pap) smear test. Persistent abnormal results require follow-up colposcopy, an anxiety-producing procedure. Anxiety and fear can interfere with adequate treatment and follow-up. Early education efforts may reduce anxiety and enhance adherence to treatment and are invaluable in halting the progression of CIN. Diagnosis and treatment, the emotional impact on women, and implications for client education are discussed. PMID- 10455566 TI - Hypertension in African-American males. AB - Hypertension is more prevalent in African-Americans than in Caucasians, and African-Americans experience more rapid progression of end-organ damage from this often "silent" disease than do whites. African-American males are particularly at risk because they often are unaware of the disease, do not receive treatment, and/or do not adhere to a treatment regimen if one is prescribed. Management of hypertension in African-American men is complex and requires consideration of the unique aspects of this special-need population. This article outlines some of the special concerns related to hypertension management in African-American men, including differences in pathophysiology, lifestyle, education, and socioeconomic status. Advanced practice nurses, with their additional knowledge and their strong emphasis on patient education and holistic care, possess the skills to competently, caringly, and creatively manage hypertension with African-American clients. PMID- 10455567 TI - Women's perceptions and concerns about menopause. AB - The purpose of this study was to explore the following questions: What are the perceptions of midlife women as to the care they receive from a variety of healthcare providers? What are the issues and concerns that influence their decision making about hormonal therapy? What strategies do women find helpful in accommodating to menopause? What are the knowledge and perceptions/role of the partner during the menopause transition? The research design for this study used a nonrandom survey. The nonrandom convenience sample consisted of 320 midlife women 46-55 years of age. Data were gathered via a mailed self-administered Menopause Questionnaire. Descriptive statistics were used to analyze the survey data. Qualitative data revealed more in-depth and personal comments from women concerning menopause. The most significant findings were that respondents (1) were in conflict about hormone therapy (HT), yet they had read extensively about menopause and used a variety of strategies to cope with symptoms; (2) reported that they received conflicting information and advice about menopause from a variety of sources and expressed concerns about not being fully informed by their healthcare providers; and (3) who received care from nurse practitioners (NPs) were quite satisfied with their care. PMID- 10455568 TI - AIDS: translating research and practice into policy. PMID- 10455569 TI - The circle of caring: a transformative model of advanced practice nursing. AB - This article presents a new and original approach to advanced practice nursing. A transformative model, the Circle of Caring is described. Using a template that builds on both nursing and medicine, this model provides a synthesized approach to practice. The traditional medical model is reviewed, as well as the evolution of the nursing process. In response to changing societal need, the scope of nursing practice is constantly challenged and expanded. New elements of practice emerge. In an effort to clarify and elaborate the current realities of advanced practice nursing, the Circle of Caring was conceptualized. Components of traditional medical practice are combined with elements of traditional nursing practice; both are situated in an elemental nursing model of caring. Although a linear representation, the Circle of Caring encompasses process pieces taken in part from a theory of caring (Boykin & Schoenhofer, 1993; Mayerhoff, 1971). An underlying purpose of this model is to highlight in a concrete way the inclusion of nursing's voice in the new and emerging--and rapidly changing--healthcare system. It is theorized that this approach to practice will yield more positive outcomes for patients, families, and communities. PMID- 10455571 TI - Profiles of editorial board members: intercontinental links. Interview by Joellen W. Hawkins. PMID- 10455570 TI - Lurkers in the water supply: using the Net to protect HIV-positive persons from cryptosporidiosis. PMID- 10455572 TI - Amoxicillin: a pharmacologic description. AB - This article discusses several pharmacokinetic aspects of amoxicillin pertinent in a maternal and pediatric population. Life stages impact both dosage requirements as well as actual pharmacokinetic properties of this antibiotic. PMID- 10455573 TI - Expanding nurse practitioner scope of practice within the Veterans Affairs Palo Alto Health Care System. AB - Healthcare priorities and use have changed dramatically over the last several years. Expansion of the nurse practitioner (NP) role has been a hallmark of the restructuring efforts, which have emphasized primary care and the use of nonphysician specialists. NPs are practicing in a wider range of settings than ever before, including acute hospital settings, outpatient clinics, and specialty services. While the Veterans Health Administration (VHA) has employed NPs since the early 1970s, their practice has been limited at many VHA medical centers. Such was the case at the Veterans Affairs Palo Alto Health Care System (VAPAHCS), when physicians produced a scope of practice (SOP) for NPs that was practice limiting. This article highlights the historical progress of NPs in defining their practice, discusses barriers to NP practice at the VAPAHCS, outlines strategies to overcome barriers, and discusses future possibilities for advanced practice nursing within the VA. PMID- 10455574 TI - Primary care screening for prostate cancer. AB - Prostate cancer is second only to lung cancer among killers of men in the United States. Researchers continue to develop tests that are more sensitive for diagnosing prostate cancer. At present, primary care assessment and evaluation of the disease are determined by physical evidence that may not be apparent and by laboratory values that may not be truly reflective of the underlying disease process. Men over the age of 40 need an annual evaluation for increased prostate specific antigen (PSA) along with a digital rectal examination. Some data suggest that the digital rectal exam and PSA levels may be insensitive indicators of prostate cancer in men with low total or free testosterone levels. The synergistic effect of testosterone on PSA could mask indicators for evaluation of prostate cancer. PMID- 10455575 TI - Antidepressant prescribing practices of nurse practitioners. AB - Depression is considered a highly treatable illness, yet often it is inadequately treated by physicians in primary care settings. The economic and human cost of inadequate treatment is well documented. The purpose of this pilot study was to explore the antidepressant prescribing practices among a convenience sample of nurse practitioners (NPs) and to use the findings to refine the antidepressant therapy questionnaire for further use. A self-administered questionnaire was completed by 26 NPs. Results of the survey revealed that NPs in primary care settings frequently treat a wide variety of common mental health problems and prescribe antidepressant medications for a number of these disorders as well as for nonpsychiatric problems. Factors that influence the NPs' decision-making process in the selection of a particular antidepressant were found to vary among the NPs surveyed. Although 65% of the NPs felt that they were sufficiently informed about the available antidepressants to select the most appropriate drug for their clients, 77% believed they needed additional education on antidepressant medications, and all but one respondent reported willingness to attend continuing educational offerings on antidepressant medications. PMID- 10455576 TI - Health problems of sheltered homeless men using a mobile health van: a 4-year study. AB - The number of homeless persons in the United States is increasing on an annual basis, with men accounting for the greatest number. Accessing health care for this group is difficult for many reasons, particularly those related to finances, transportation, lack of insurance, and transiency of lifestyle. Consequently, data regarding homeless men's health problems/concerns are difficult to obtain, and often the information reported is only for acute care services provided by emergency rooms and clinics. The purpose of this study was to determine the demographic characteristics and health problems/concerns of sheltered homeless men using the services of a mobile van for health care over a 4-year period. A total of 2,086 records, representing 1,171 men aged 19 and over, were retrospectively reviewed to determine demographic and health characteristics (age, sex, self-reported medical and addiction histories, and self-reported medication use). Data on presenting concerns, diagnoses, visit patterns, treatments, medications and teaching provided were also collected and analyzed. Recommendations to improve the quality, continuity, and follow-up of care for homeless men are included. PMID- 10455577 TI - Women's health: where are nurse practitioner programs headed? AB - Until quite recently, the discipline of women's health has been restricted to reproductive health. While graduate nurse practitioner (NP) programs have included issues across the life cycle and health promotion in their curricula, there is growing emphasis on primary care of women. Graduate nursing programs that offer women's health and obstetric/gynecologic NP tracks were surveyed in 1996 to elicit content in the curriculum and to ascertain whether changes in requirements for certification were impacting programs. Programs listed in the 1994 National Organization of Nurse Practitioner Faculties directory (NONPF) were surveyed. Twenty-eight of 31 programs responded; results were tallied by frequency distribution. Results documented differences in programs, suggesting that a clear definition of women's health advanced practice is needed. The author recommends a national dialog to explore advanced practice in women's health and asks if there should be two types of NP programs--primary care of women (a generalist NP) and reproductive health (a specialist NP)--and how NP programs and credentialing bodies should differentiate graduate programs as well as content for certification exams. PMID- 10455578 TI - Independent, autonomous, or equal: what do we really want? AB - This article will explore the development of terms such as autonomous and independent in the national and international nursing literature, with particular reference to the development of the nurse practitioner (NP)/advanced nursing role. It will examine the complexity of power relationships in health care and explore the apparent dissatisfaction of nurses with their status, as demonstrated by such issues as the proliferation of nursing ethics and the problems surrounding recruitment and retention. It will discuss why the development of advanced practice roles has been greeted with such suspicion by some medical organizations, and what barriers and benefits there are to collaborative practice. It will explore the relationship between the bodies of knowledge of medicine and nursing, and the perceived blurring of roles that advanced practice can potentiate. Finally, it will make some suggestions for intelligent and harmonious coexistence, which would refocus medical and nursing practice on patient care, rather than on "bunker dwelling." PMID- 10455579 TI - NPACE nurse practitioner practice characteristics, salary, and benefits survey. AB - The purpose of this article is to present the results of a survey of practice characteristics, salary, and benefits of 1,763 nurse practitioners (NPs) from the eastern United States who attended national NP conferences in Chicago, Illinois, and Boston, Massachusetts, in the fall of 1996 and in San Diego, California, and Orlando, Florida, in the spring of 1997. Specific data are presented on the demographics of the population; practice characteristics and responsibilities; benefits for full- and part-time employees; and salary by region, years of practice, type of certification, and location of the practice. The salary data were compared with the 1995-1996 NPACE practice characteristics, salary, and benefits survey for the eastern United States. PMID- 10455580 TI - Nurse practitioners in the United Kingdom: some considerations of the literature. AB - This article identifies issues leading up to changes in British health care and addresses the literature surrounding specialist and advanced nursing practice. American literature on advanced nursing practice includes specialist nurses and nurse practitioners (NPs), whereas the British debate isolates specialist and advanced nursing practice. The credentialing, regulation, and titling of advanced nursing practice in the United Kingdom is in abeyance while discussions address how specialist practice can encompass both clinical nurse specialists and NPs. Care is therefore required when making cross-cultural comparisons of advanced nursing practice. The problems associated with lack of clarity in the preparation and assessment of the advanced nurse practitioner are highlighted, and questions are raised about the nonacademically prepared experienced nurse clinician. Issues of NP accountability and responsibility are addressed. PMID- 10455581 TI - Searching MEDLINE free on the Internet using the National Library of Medicine's PubMed. AB - Searching MEDLINE, the National Library of Medicine's premier bibliographic database covering the fields of medicine, nursing, dental medicine, the healthcare system, veterinary medicine, and the preclinical sciences, is now available free on the Internet through PubMed. This retrieval system allows users of all skill levels to obtain important medical information. Additionally, through the National Network of Libraries of Medicine (NN/LM) program, a division of NLM, user support in the form of phone, e-mail, and training is available free of charge. Those interested in having a training session for PubMed at their facility may contact the NN/LM to make arrangements. PMID- 10455582 TI - Profiles of editorial board members: from Jamaica to Vermont. Interview by Joellen W. Hawkins. PMID- 10455583 TI - Moving research utilization into the millennium. PMID- 10455584 TI - Research utilization: current issues, questions, and debates. PMID- 10455585 TI - Transferring public-health nursing research to health-system planning: assessing the relevance and accessibility of systematic reviews. AB - A descriptive study was designed to gain an understanding of the research needs, perceptions of barriers to research utilization, and attitudes towards systematic reviews of decision-makers in public health at the level of systems planning. Public-health consultants and managers in Ontario were surveyed about barriers to research utilization and awareness of and attitudes towards systematic reviews as a method of/vehicle for research transfer. Access to 5 completed reviews was provided in summary, abstract, and full form, and on diskette, hard copy, and Internet. A follow-up survey at 3 months assessed use, relevance, application, and further dissemination of the reviews. A total of 242 people in positions of public-health policy and decision-making participated. Respondents reported a great, largely unmet, need for research evidence. They viewed systematic reviews as likely to overcome the barriers to research use related to critical appraisal, time, timeliness, availability, cost and credibility, but not the barriers related to policy climate, authority, or implementation resources. Three months after requesting a review, 93% said they would follow it up; 91% remembered receiving it, and 71% of these had read it while 23% stated it played a part in program planning or decision-making. PMID- 10455586 TI - Evidence-based pressure-ulcer practice: the Ottawa model of research use. AB - Developing evidence-based nursing practice among diverse health-care settings is a particular challenge in the face of current health-care restructuring. This paper describes application of the Ottawa Model of Research Use (OMRU) to increase evidence-based practice across 3 health-care settings during a time of multiple restructuring changes. The initiative was part of a provincial demonstration project to develop centers of nursing excellence with a view to improving continuity of care across the health continuum. Three Ottawa health care agencies formed one of 4 participating sites in the Province-Wide Nursing Project (PWNP), a 3-year initiative funded by the Ontario Ministry of Health. The goal of the Ottawa-Carleton site was to increase evidence-based decision-making with a focus on pressure ulcers. The barriers and supports encountered, and the strategies used, in striving to meet this goal in a community-care, tertiary care, and long-term-care setting are described. Multiple research transfer approaches were used, with an emphasis on education. Implementor consensus and achievements of the project support the OMRU's utility as a guide to implementing research findings in these health-care settings. PMID- 10455587 TI - Mapping the research utilization field in nursing. AB - The recent increase in interest in the field of research utilization, often embedded in the notions of evidence-based practice, presents a rich opportunity to advance the field in nursing. While an extensive literature on the subject exists in nursing, close examination reveals that much of it is opinion and anecdotal literature, and that sustained and programmatic theory building and testing in this field has been sporadic at best. This article maps the field of research utilization, proposing that we focus on major areas of inquiry: scientific, historical, and philosophical foundations, synthesis, determinants, policy, interventions to increase research utilization, and outcomes. In so doing, alternative ways of viewing and conceptualizing this field are possible. In conducting the kinds of studies and supporting the kinds of programs identified in this map, nursing, in collaboration with appropriate partners, can significantly advance the field of research dissemination and utilization studies and practice at many levels in the health system. PMID- 10455588 TI - The use of metaphor graphics to depict sleep research results. PMID- 10455590 TI - Dissemination: current conversations and practices. PMID- 10455589 TI - Evidence that informs practice and policy: the role of strategic alliances at the municipal, provincial, and federal levels. PMID- 10455592 TI - Evaluation of three brands of tympanic thermometer. AB - Infrared tympanic thermometers (ITT) have many documented benefits, including speed, ease of use, and noninvasiveness, to support their use in emergency departments (ED) and intensive care units (ICU). However, concerns have been raised about the accuracy of temperatures reported by ITT. This study was conducted to evaluate the accuracy of 3 brands of ITT, compared to rectal and pulmonary artery thermometers, in ED and ICU settings. Results indicated adequate specificity for all 3 ITT in ED (range = 0.9242 to 1.0000) and ICU (range = 0.9737 to 1.0000), but unacceptable sensitivity in ED (range = 0.5455 to 0.8000) and ICU (range = 0.0000). Further analyses indicated highly variable ITT-reported temperatures. In ED, ITT temperatures were, on average, 0.3684 degree C lower, and could be expected to vary by more than 2 degrees C from the actual temperature as reported by rectal thermometer. In ICU, the average reported temperature was similar to the actual pulmonary artery temperature (0.0259 degree C lower), but again could vary by more than 2 degrees C. Recommendations for nursing and education are discussed. PMID- 10455591 TI - Exploring the experience of type 2 diabetes in urban aboriginal people. AB - The experience of diabetes among urban Aboriginal people (status and non-status Indians) was explored through a qualitative study. Because researchers have focused almost exclusively on Aboriginal people living on reserves or in isolated rural communities in Canada, this study conducted face-to-face interviews with participants (n = 20) living in the city of Winnipeg, Manitoba. The data generated 3 themes: diabetes as an omnipresent and uncontrollable disease; beyond high sugar: diabetes revealed in bodily damage; and the good, the bad, and the unhelpful: interactions with health-care providers. Findings from this study and previous research support the existence of a pan-Aboriginal model of diabetes. This contemporary cultural stance appears to transcend geography and has implications for the prevention and treatment approaches used in programs and health services for Aboriginal people living with diabetes. PMID- 10455593 TI - [An investigation on the family support of discharged acute myocardial infarction patients]. AB - Acute myocardial infarction (AMI) is a kind of diseases. The rehabilitation of AMI patients is affected by many psychosocial factors, the family support is one of these factors. In this research, two groups of patients were interviewed by the author with the Family APGAR Questionnaire, one group was of the patients who discharged 4-6 years ago, the other group was of the patients who discharged 5-7 months ago. The result was that the family support of five-year group was better than that of half-year group. So, the education for patient's family numbers was very important and should be thought highly by nurses. PMID- 10455594 TI - [An investigation on anxiety and coping patterns of patients under cancer chemotherapy and the strategies]. AB - 100 cases of patients under cancer chemotherapy were investigated by using Self Rating Anxiety Scale (SAS) and questionnaire on coping patterns. The results showed that scores of SAS were significantly higher than that of the normal population. The state of the anxiety reached 45% among the patients. The patients' coping patterns tended to be passive. Linear analysis revealed a negative correlation between the coping patterns and the scores of SAS. It is recommended that cognitive behavioral therapy can be used to relieve the anxiety of patients under cancer chemotherapy. PMID- 10455595 TI - [Study on measuring blood pressure and heart rate in patients taking antipsychotics]. AB - To decrease the prevalence of orthostatic hypotension in patients taking antipsychotics 62 pchizophrenics' blood pressure and heart rate were measured at different posture. The data showed that the systolic pressure, diastolic pressure and heart rate in supine position were statistically different from those at standing position (P < 0.01). In standing position the heart rate was 41/min higher, the systolic pressure 1.32 kPa lower and the diastolic pressure 1.2 kPa higher than those in supine position. The systolic pressure of 5 cases were normal in supine position and under 12 kPa in standing position. It is concluded that the heart rate and blood pressure should be measured both in standing and supine position in patients taking antipsychotics, so as to prevent the onset of orthostatic hypotension. PMID- 10455596 TI - [Rehabilitative nursing after flex tendon repair of the hand]. PMID- 10455597 TI - [Nursing care of complications after hydrocephalus shunts]. PMID- 10455598 TI - [The effects of intravenous nitroglycerin or sodium nitroprusside infusion on the blood pressure in patients with acute myocardial infarction]. PMID- 10455599 TI - [Clinical observation and nursing of nitrous oxide inhalation to ease delivery pain]. PMID- 10455600 TI - [Nursing care of hypertrophic rhinitis treated by microwave thermocoagulation]. PMID- 10455601 TI - [Psychological analysis and nursing of heroin addicts: report of 1000 cases]. PMID- 10455603 TI - [Enhancing the learning of the nursing process]. PMID- 10455602 TI - [Coordination of transnasal, transethmoid, sphenoid decompression of the optic nerve canal]. PMID- 10455604 TI - [The application of the "introversion type" motivation method in nursing management]. PMID- 10455605 TI - [Management of continuing nursing education and scoring system]. PMID- 10455606 TI - [Study on phlebitis caused by intravenous drips]. PMID- 10455607 TI - [Study of hemolysis caused by quantitative automatic vein hemostix]. PMID- 10455608 TI - [Verbal communication in the nurse-patient relationship]. PMID- 10455609 TI - 'Getting our act together' in nursing. PMID- 10455610 TI - The good, the bad and the relative, Part Two: Goodness and the criterion problem in qualitative research. AB - This (part two) paper finds 'the problem of the criterion' at the heart of disputes about what constitutes goodness in qualitative research, an ancient philosophic conundrum as to how best represent reality. Ways around the problem are considered, including conceiving criteria as open-ended 'lists', and 'enabling conditions'. Discussion principally concerns the impact of postmodernist thinking on the topic, and how qualitative researchers might usefully juxtapose the rationality of a modern world (in which notions such as reliability and validity are prized) with a mounting postmodern sensibility that acknowledges irrationality, fragmentation, and uncertainty. Part one of the paper traced efforts to define 'goodness' in qualitative research within various fields, including nursing. Disputes were found to centre on how the traditional concepts of reliability and validity related to qualitative research. In reviewing various sets of criteria of goodness, these concepts were consequently conceived as being championed, translated, exiled, redeemed, and surpassed. PMID- 10455611 TI - Moral distress, advocacy and burnout: theorizing the relationships. AB - Burnout is a major contributor to shortages of experienced nurses. The research literature shows a correlation between moral distress and burnout in critical care nurses. This paper reports on part of an interpretive interactionist study concerning nurses' experiences of moral distress which prompted attempts to advocate for vulnerable patients. One critical incident is used as an example of the qualitative findings of the study. In this paper, we theorize about what happened when nurses advocated for their patients by challenging medical treatments which the nurses believed to be both inappropriate and to contributing to patient suffering. When attempts at advocacy were unsuccessful, the nurses experienced intensified moral distress, frustration and anger. Being an unsuccessful advocate resulted in nurses being relocated within the hospital, nurses being scapegoated and/or burning out. The theoretical links which this paper makes between advocacy, moral distress and burnout are supported by empirical data from the study. PMID- 10455612 TI - Registered nurses' perceptions of teaching: constraints to the teaching moment. AB - Nursing literature indicates that patient teaching is an integral role function of the nurse. While some authors suggest that patient teaching is an essential part of nursing care, others argue that nurses are not adequately prepared to assume the role of teacher. This study explores the context within which nurses engage in patient teaching, revealing tensions experienced on a day to day basis within the practice setting. The principles of teaching and learning are also explored, raising questions about the extent to which nurses utilize these principles to guide the teaching moment. It is asserted that nurses find it difficult to engage in activities which are pivotal to the provision of quality patient education. Whether this is because they have little awareness or understanding of the principles of education, or whether this is related to the milieu in which teaching takes place, is explored. PMID- 10455613 TI - Pain in children: qualitative research of Finnish school-aged children's experiences of pain in hospital. AB - The purpose of the study was to describe school-aged children's experiences of pain in the hospital. Data were collected from 20 children aged between seven and 11 years of age who were inpatients in the pediatric wards of the University Hospital, Oulu, Finland. Each of the children wrote of their experiences of pain in the hospital, and they were interviewed on the basis of this information. The data were analyzed inductively using content analysis. On the basis of the analysis, the situations that caused pain to all children in the hospital were found to be procedures connected with needles. The children described their experiences of pain as both physiological (e.g. poking, stinging, aching) and psychological feelings (unpleasant feelings, terror, anxiety, and fear). In addition, methods of relieving pain (coping mechanisms, help from the medical staff, parental presence, and previous experiences of similar situations) were acknowledged. The results indicate that school-aged children (aged 7-11 years) are able to describe their pain experiences, which should be considered in assessment and treatment of children's pain in nursing practice. PMID- 10455614 TI - Registered nurses' descriptions of their health assessment practices. AB - The delivery of health-care services in Australia is undergoing many changes. The move towards home-based acute care has precipitated a review of many nursing care practices. An important consequence of these changes is the need for registered nurses to be adequately equipped to conduct systematic health assessments. This descriptive study used a questionnaire designed to elicit short-answer responses in order to investigate how registered nurses described their health assessment practices and what type of data they collected. The findings indicate that respondents possess divergent conceptualizations of health assessment, and that they predominately use health assessment data to support medical therapies. Discussion focuses on the implications of these findings for nursing practice. PMID- 10455615 TI - Selected factors influencing self-concept among hospitalized Chinese school-age children with a chronic illness. AB - A correlational descriptive study was conducted to investigate self-concept and selected influencing factors among hospitalized Chinese school-age children with a chronic illness. The purposive sample was composed of 122 school-age children with chronic illnesses, such as nephrotic syndrome, leukemia, and congenital heart disease, who were admitted to three major teaching hospitals in Shanghai City. The children's medical records, demographic forms, and the Piers-Harris Self-concept Scale (PHSCS) were used to collect the data. Descriptive statistics, and stepwise multiple regression were used to analyze the data utilizing the SPSS program (SPSS Inc, Chicago, IL, USA). The results of this study showed that the majority of hospitalized Chinese school-age children with a chronic illness had at least an average level of self-concept. Academic achievement (grade point average) was a strong predictor of self-concept, as are most of its subconcepts, including social behavior, academic competence, and physical appearance and attributes. Age was a significant predictor of overall self-concept, social behavior, and popularity subconcept. Sex was a significant predictor of self concept subconcepts such as social behavior, anxiety, and popularity. Duration of illness was a significant predictor of physical appearance and attributes. In addition, the type of illness was a significant predictor of popularity. It can be concluded that the importance of school education should be emphasized by hospital and school nurses for the purpose of helping such children build a positive self-concept concerning their age, sex, academic achievement, type of illness, and duration of illness. Recommendations for further research have been suggested. PMID- 10455616 TI - Ovarian cancer: an update for nursing practice. AB - A diagnosis of ovarian cancer is a crisis for any woman. Ovarian cancer has the highest mortality of all the gynaecological cancers. In addition to the very real threat of death, such a diagnosis raises critical issues concerning femininity, motherhood and sexuality. The high mortality associated with this disease is due (in part) to problems associated with early diagnosis. Women frequently experience non-specific symptoms, which may be attributed to other, less alarming causes. The lack of specific symptoms experienced by women, as well as the lack of effective mass screening strategies, make early detection difficult. This makes it crucial that nurses involved in the care of women be alert to the insidious and non-specific onset of this serious condition. PMID- 10455617 TI - Almost 80. Aged population: expectations and roles. PMID- 10455618 TI - The Behavioural Status Index: therapeutic assessment of risk, insight, communication and social skills. AB - An overview is given of the Behavioural Status Index (BSI), a developing classification instrument offering practical approaches to assessment and therapy surrounding social 'risk'. The approach hypothesizes that social 'risk' presented by a patient tends to correlate inversely with his/her degree of personal insight and capacity to perform well in key communicative and social skills, though no causal claims are made. Evidence exists to suggest that personal insight and communicative and social skills deserve serious consideration in a therapeutic approach to violent and dangerous behaviours. 'Risky' behaviours, as operationalized in the 'risk' subscale of the BSI, insight into the self and its activities, and communicative and social skills, may provide three critical foci for treatment planning in high security psychiatric care. A hypothetical linked factorial structure is proposed. PMID- 10455619 TI - Waiting for harm: deliberate self-harm and suicide in young people--a review of the literature. AB - Mental health professionals, particularly nurses, practicing in child and adolescent psychiatry are key players in the continued development of expertise, awareness and understanding of deliberate self-harm (DSH) and suicide in young people. Enhancing this knowledge and the associated skills will contribute to effective, therapeutic relationships with individuals and their families. The purpose of this paper is to review the current literature surrounding the issue of deliberate self-harm and suicide in young people. The paper will address national and international studies selected from a variety of child and adolescent and general psychiatric literature. The research presented highlights important issues to be addressed in practice. In particular, the evidence supports the influence of mental illness in DSH and suicide, particularly depression and substance abuse. However, a clearly important factor is an individual's experience of family and social life. The literature is considered in the light of relevant governmental policy documents relating to DSH and suicide. The future role of the mental health nurse in child and adolescent psychiatry in the United Kingdom (UK) is discussed in the contexts of family work and assessment. PMID- 10455620 TI - Psychological needs of patients with HIV disease: reviewing the literature using Nichols' (1985) Adjustment Reaction Model as a framework. AB - Since the first manifestations of HIV infection presented in the United Kingdom in the early 1980s, there has been a dramatic increase in the incidence of HIV infection and a proliferation of diseases associated with HIV infection. HIV infection is now associated with physical, psychological, mental and neurological conditions. Many of these conditions are serious and pose a major threat to physical and mental health, with rapid deterioration of health status necessitating multiple hospital admissions to acute hospitals. The majority of people hospitalized with HIV diseases are currently being cared for on general wards. While physical needs are often well met, psychological needs are to a certain extent neglected. The literature cites numerous cases of neglect, discrimination and threats to confidentiality by health professionals, including nurses. Such psychological needs, emanating out of hospitalization, compound existing psychological needs. Nichols' (1985) Adjustment Reaction Model describes a number of stages patients may go through during the course of their illness, and to some extent the patient's psychological needs can be traced throughout the trajectory of the disease. The purpose of this paper is to review the literature and present a theoretical argument, using Nichols' model as a framework, on psychological needs of patients with HIV diseases, and attempt to outline to what extent psychological needs of patients with HIV are met in institutional care. The authors recommend further research to determine nurses' perception of the importance of meeting psychological needs of patients with HIV disease. PMID- 10455621 TI - An exploratory account of registered nurses' experience of patient aggression in both mental health and general nursing settings. AB - Until relatively recently, aggression and violence in nursing has been an area of little discussion (Poster & Ryan 1993). Today, concerns are rising about escalating levels of violence towards nurses (Wykes 1994, Whittington 1997). In response, this paper explores registered nurses' experiences of patient aggression encountered in the acute inpatient general and mental health settings. Critical incidents from registered nurses in both areas are examined and analyzed in relation to existing literature. The aim of the research is to ascertain if differences exist or similarities prevail. Findings reveal that the two areas have similar problems in terms of types of aggression. Verbal and 'minor' types of aggression are the most problematic. Also, the 'biomedical model' of care is evident in both settings and possibly underpins chosen approaches to aggression management. The repeated use of chemical and physical restraint is apparent. The main comparison identified between the two groups relates to nursing control over situations involving violent patients. Mental health nurses seem to consistently take control of aggressive situations whilst general nurses tend to rely more heavily upon the input of others (medical staff, mental health teams and the police) when intervening. These two nursing specialties therefore have much to learn from each other in terms of nursing experience and possible future approaches to the management of aggressive patients. PMID- 10455622 TI - Psychiatric nursing for promotion of mental health and prevention of depression in the elderly: a case study. AB - The aim of this case study was to describe and analyse the basic pre-conditions for an intervention programme focusing upon mental health promotion and prevention of depression in an elderly community group and to discuss the psychiatric nurse's community-oriented health promotion work. The intervention programme, which was led by a psychiatric nurse, was built on intersectorial co operation between public and voluntary organizations, development of social networks to promote social support, and target group participation. The nurse's function in the community-oriented work was to act at the individual, group and society level, as well as to mobilize resources among individuals and organizations, in order to create a favourable interaction, resulting in health and empowerment for the individual. Pre-conditions for leading this work were: knowledge of health and mental all health among the elderly, investigation methodology, knowledge about the local society, as well as pedagogical, supervisory and social competence. PMID- 10455623 TI - Research utilization and attitudes towards research among psychiatric nurses in Northern Ireland. AB - Evidence-based practice depends on a number of factors including practitioners' attitudes towards, and knowledge of, research, the availability and access to relevant research, adequate resources and managers' support. Yet little is known about these issues with regard to psychiatric nurses. The aim of this study was to survey psychiatric nurses on their attitudes to research, their perceptions of their use of research and other research-related activities. This paper reports data collected from a convenience sample of 236 nurses from the six main psychiatric hospitals and from the psychiatric wards of six general hospitals. The results show that, while they report positive attitudes towards research, their perception of their use of research in practice indicates that evidence based practice is far from being realized. There is also some evidence to suggest that psychiatric nurses' reported extent of research utilization and frequency of reading research literature in this study are lower than those of general nurses. The implications of these and other findings are discussed. PMID- 10455624 TI - Education for community psychiatric nurses: content, structure and trends in recruitment. AB - Under the auspices of the Standing Advisory Group for Community Psychiatric Nursing Education, annual surveys of the leaders of the UK's post-registration courses for community psychiatric nurses (CPNs) have taken place since the 1987 88 academic year. In this paper, findings are reported from the 1997-98 survey. Thirty-two of the 39 course leaders running post-qualifying programmes for CPNs (English National Board course 812 or equivalent, and Specialist Practitioner courses) in the 1997-98 academic year responded to a nine-page postal questionnaire. Responses suggested that the majority of courses for CPNs are now offered at first degree level, with most students completing their programmes of study and clinical practice in one full-time academic year. Most courses appeared to include education in key areas of specialist content pertinent to contemporary CPN practice. Examples included: evidence-based psychosocial interventions; collaborative working with users of mental health services; inter-professional issues; cognitive-behavioural interventions; reflective practice and clinical supervision; and medication-related issues. Overall, however, courses for CPNs appeared to be characterized by considerable variations in specialist content. Possible explanations for this are offered, along with suggestions for future research. PMID- 10455625 TI - The role of reflection in the development of practice theory: a case study. AB - Many authors view practice theory as the strongest form of theory for an applied discipline like nursing. It is based upon the idea that theory emanates from practice, is strengthened and is returned to inform practice. The first part of this paper is concerned with identifying the contribution that practice theory can make to patient care. While there are many positivistic ways of generating practice theory, one method which is getting increasing attention is reflection. In this paper, Boud et al.'s framework for reflection is used to identify phenomena related to the empowerment of patients and staff in psychiatric hospitals (Boud et al. 1985). A real life case study is described and concepts and beginning propositions are uncovered. Conclusions are drawn which from the basis for theory development and testing. PMID- 10455626 TI - Emotional competence in professional communication. PMID- 10455627 TI - Honey and wound bacteria. PMID- 10455628 TI - Leg ulcer care in the community. PMID- 10455629 TI - The use of honey as an antiseptic in managing Pseudomonas infection. AB - A laboratory study was undertaken to extend existing knowledge about the effectiveness of the antibacterial properties of honey against pseudomonads. To date, sensitivity testing has used non-standardised honeys, which may vary greatly in their antibacterial potency. Pure cultures of Pseudomonas spp, isolated from swabs from 20 infected wounds, were inoculated on the surface of nutrient agar plates containing various concentrations of honey in the medium. Two types of honey were used, a manuka honey and a pasture honey, each selected to have antibacterial activity close to the median for each type. The minimum inhibitory concentration of the manuka honey for the 20 isolates ranged from 5.5 8.7% (v/v) (mean 6.9% (v/v), standard deviation 1.3). The minimum inhibitory concentration of the pasture honey for the 20 isolates ranged from 5.8-9.0% (v/v) (mean 7.1% (v/v), standard deviation 1.0). Honeys with an average level of antibacterial activity could be expected to be effective in preventing the growth of pseudomonads on the surface of a wound even if the honey were diluted more than ten-fold by exudation from the wound. PMID- 10455630 TI - A follow-up study of leg ulcer patients in south Stockholm. AB - The objective of this study was to follow up patients with leg ulcers, previously identified in an epidemiological study in South Stockholm. Rate of healing, non healing, recurrence, amputation and mortality were recorded. A total of 254 patients were identified for follow-up after 18 months, 217 of whom were included. The use of a questionnaire revealed that, during this period, 43 (20%) of the patients had died; of the remaining 174 patients, 61% had healed, 29% were unhealed, 8% had recurring ulceration after healing; 2% had required amputation. Venous aetiology was found to be more common than arterial aetiology. However, among those who had died, arterial disease was found to be more common than venous insufficiency. A further objective was to investigate 21 patients with non healed venous leg ulcers who had been treated in community care. They were assessed by means of a standardised form and given a triple-layer bandaging treatment for a three-month period. Two ulcers had healed and nine ulcers had improved and were healing. In the group of patients with healed or improved ulcers, five had normal mobility. PMID- 10455632 TI - Dermatological conditions and HIV disease. PMID- 10455631 TI - The use of larval therapy in wound management in the UK. AB - This study identifies hospitals and institutions in the UK using larval therapy, and determines how this therapy is clinically managed in 23 of them. A qualitative approach was adopted, with the collection of documentary evidence and data from semi-structured interviews. Larval therapy is currently used in over 350 hospitals and institutions in the UK in the treatment of a variety of wound types. It was evident from the findings that it is generally used as a last resort. However, in two hospitals, this treatment method is part of the hospital's wound management policy. Nurses were able to identify wounds that have a better response to larval therapy and recognise symptoms that may occur during treatment. A number of wound types were found to be 'difficult' with regards to larvae application, and nurses had developed their own practices involving the application of dressings and the length of time larvae were left on wounds. Patients receiving this treatment reported few symptoms. It was claimed that larval therapy is particularly effective against methicillin-resistant Staphylococcus aureus (MRSA). PMID- 10455633 TI - Challenging the pressure sore paradigm. AB - This study determines the effectiveness of a new low-unit-cost support system in patients at very high risk of developing pressure sores. In a prospective randomised controlled trial, a low-pressure inflatable mattress and cushion system (Repose) was compared to a dynamic support mattress (Nimbus II) used in conjunction with an alternating-pressure cushion (Alpha TranCell) in 80 patients with fractured neck of femur and high scores on a pressure sore risk assessment scale. All patients received best standard care, including turning at regular intervals. Skin condition was assessed in 17 locations on admission, preoperatively, and seven and 14 days postoperatively. No difference was found between the groups in skin condition or the occurrence and severity of pressure sores at any time point. PMID- 10455634 TI - The prevalence of pressure damage in acute care hospital patients in the UK. AB - This paper aims to provide information on the prevalence of pressure damage in UK hospital patients since 1992. A survey method and data handling service provided by a medical device company was used to set targets and monitor trends. The results of these surveys provide evidence that over a six year period, the prevalence of pressure damage in this population in the UK has shown a significant reduction. PMID- 10455635 TI - Pharmacological treatments for venous leg ulcers. PMID- 10455636 TI - The causes and prevention of maceration of the skin. PMID- 10455637 TI - The use of hydrocolloids in the treatment of diabetic foot. PMID- 10455639 TI - [Is there a right to a quick easy death?] [In Process Citation] PMID- 10455638 TI - Ulcer care in the Middle Ages. PMID- 10455640 TI - [Patients with epilepsy: the person, not the disease, is the focus]. PMID- 10455641 TI - [So talks may build bridges...]. PMID- 10455643 TI - [Creutzfeldt-Jacob syndrome: risk for patient and health personnel?]. PMID- 10455642 TI - [Caring for abused women: the male nurse as a countermodel]. PMID- 10455644 TI - [The sparrow at the bedside]. PMID- 10455645 TI - [Oil in the bath water]. PMID- 10455646 TI - [Is it really so hard to move?] [In Process Citation] PMID- 10455647 TI - [Anthroposophical care. A vision of totality]. PMID- 10455648 TI - [Fluid intake in the aged. Always one more glass!]. PMID- 10455649 TI - [Drug addiction and pregnancy. The liaison nurse's role]. PMID- 10455650 TI - [An efficient method to reduce odors]. PMID- 10455651 TI - [What are the prospects of teaching by concept?]. PMID- 10455652 TI - [Patients' rights. Respect for the private sphere and the nursing mandate]. PMID- 10455653 TI - [International council of nurses. The year of the centennary has begun] [In Process Citation] PMID- 10455654 TI - International centres for nursing ethics. PMID- 10455655 TI - Acting from the virtue of caring in nursing. AB - The author challenges the recently argued position of Helga Kuhse that caring is merely a preparatory stage to moral action and that impartial, principled thinking is required to make action moral, by suggesting a notion of caring as virtue. If caring is a virtue then acting from that virtue will be acting well. Acting from the virtue of caring involves eight features, which include not only that of being sensitive to, and concerned about, the patient, but also that of being aware of, and sensitive to, the relevant ethical principles. In this way, caring is seen as an overarching quality that gives action its moral character. The moral character of an action does not derive only from its having been performed in the light of principles. PMID- 10455656 TI - Educating for interprofessional collaboration: teaching about values. AB - Effective interprofessional collaboration depends upon establishing understanding that respects differences in values and beliefs, and thus differences in response to the multiplicity of patient/client/user needs. To facilitate the latter, this article suggests that health and social care students need a formal knowledge of the meaning of values and the varieties of systems within which values are expressed. Students need especially to understand the genesis of their own professional value system and to recognize the gap that inevitably develops between the values of the professional and those of the society within which a professional may function. The conceptual framework that underpins the approach to teaching values to health and social care professionals advocated here is derived from key concepts identified from the literature relating to education for, and participation in, a democratic, multicultural, multifaith society. These are: tolerance, compromise and education for dialogue. Finally, it is suggested that professional educators must take seriously the tasks of educating for professional pluralism. PMID- 10455657 TI - Care as a moral attitude in nursing. AB - The concept of care can be explained in various ways, and it can present a different meaning to each person. Nurses are increasingly aware that good nursing care consists of 'more' than the competent performance of a number of caring activities. For many nurses it is less clear what this 'more' means and what importance it has in nursing. This article will develop a view concerning care considered as a moral attitude. It is argued that care can be considered as a foundational normative concept in the ethics of the nursing profession. The aim is to clarify that nurses do not derive their specific caring identity just from the set of tasks that they perform but also from the way in which they commit themselves to the caring process. PMID- 10455658 TI - Ethical considerations in a grounded theory study on the dynamics of hope in HIV positive adults and their significant others. AB - The purpose of this article is to describe and reflect ethical challenges in a grounded theory study on the dynamics of hope in HIV-positive adults and their significant others. It concentrates on the justification of a research problem, sensitive research and the relationship between the researcher and the participants in data collection. The basis of ethically sound nursing research on the dynamics of hope in these two vulnerable groups lies in the relationship between the researchers and the participant. However, it is also obvious that the content, the process, the methods used and the ethics of the study cannot be divorced from this relationship. In conducting grounded theory research on the dynamics of hope in this research population, the researcher has to consider the surrounding world, that is, the reality in which these people live in hope or despair. PMID- 10455659 TI - Allocating scarce resources in a publicly funded health system: ethical considerations of a Canadian managed care proposal. AB - In the Canadian health care system, the Government is responsible for allocating scarce resources in a fair and equitable manner. A proposal to implement managed care as a method of reimbursing physicians in Alberta, Canada, needs careful ethical consideration, because physicians are not well prepared, and should not be asked, to make the resulting difficult allocation decisions. The Government must continue to be held responsible for ensuring that all citizens have equal access to necessary medical services, and we must find ways to encourage the public to become more involved in deciding how resources are best allocated. Health professionals other than physicians must take an interest and enter into this debate. PMID- 10455660 TI - 'We went through psychological hell': a case report of prenatal diagnosis. PMID- 10455662 TI - Frank talk. Interview by Adele Waters. PMID- 10455663 TI - An exceptional nurse. Interview by Julie McCaughan. PMID- 10455664 TI - Life at the border. PMID- 10455665 TI - Total gorilla care. Interview by Charlotte Alderman. PMID- 10455666 TI - Energy field. PMID- 10455667 TI - No holds barred. PMID- 10455668 TI - With patient attached. PMID- 10455669 TI - Close encounters. PMID- 10455670 TI - Personnel non grata. PMID- 10455671 TI - The Queen's Nursing Institute. AB - This report describes the work of the Queen's Nursing Institute, promoting best practice in community care for more than a century. PMID- 10455672 TI - Teaching in practice. AB - This article describes an action research project designed to develop a teaching programme which would enable students to bridge the theory-practice gap they experienced while on placement in a gynaecology unit. PMID- 10455673 TI - Role of the community nurse in caring for people with depression. AB - Depression is often seen as a defect of character, rather than a disorder which can be treated. Elizabeth Armstrong outlines how nurses working in primary care can effectively recognise and refer patients or clients with depression. PMID- 10455674 TI - Stamp out stress. PMID- 10455675 TI - Professional development. PMID- 10455676 TI - Starting your career. PMID- 10455677 TI - Sick and tired of shortages. PMID- 10455678 TI - Picking up the pieces. Interview by Kate Williams. PMID- 10455679 TI - Retaining a sense of proportion. PMID- 10455680 TI - The downfall of Norplant. PMID- 10455681 TI - In a class of their own. PMID- 10455682 TI - Crossing the divide. PMID- 10455683 TI - Diary of a returner. PMID- 10455684 TI - Fertility lottery. PMID- 10455685 TI - Value for money. PMID- 10455686 TI - Oral care. PMID- 10455687 TI - The careers of enrolled nurses following completion of conversion courses. AB - This article examines the extent of academic and professional development of enrolled nurses who have completed a conversion course. PMID- 10455688 TI - Non-judgemental care as a professional obligation. AB - In this article, Andrew Koh argues that non-judgemental care is a professional obligation with clear ethical and legal implications for nurses. PMID- 10455689 TI - Written disclosure: a way of coping with traumatic stress. AB - Verbal debriefing after a traumatic incident may interrupt the natural adaptive processes that help people to deal with such incidents. This article examines the effectiveness of written interventions to help people experiencing distress after trauma experienced at work. As an alternative to critical incident debriefing, the authors recommend a staff education programme in tandem with written disclosure. PMID- 10455690 TI - A regional nurse-led osteoporosis clinic. AB - This article describes the work of a nurse-led osteoporosis clinic and its perceived benefits to both patients and staff. PMID- 10455691 TI - Managing the child with gastroenteritis. AB - This article discusses the treatment of children with gastroenteritis and takes account of the current evidence based guidelines for management. PMID- 10455693 TI - Educational funding. Where to look. PMID- 10455692 TI - A systematic approach to antipsychotic drug treatment. PMID- 10455694 TI - Nurse managers in A&E. Interview by Frances Pickersgill. PMID- 10455695 TI - Administration of an enema. PMID- 10455696 TI - The proper focus of primary care is not treatment but prevention. PMID- 10455697 TI - Why we must help this child. PMID- 10455698 TI - Is it possible to have a truly open debate on the rationing of health care resources. PMID- 10455699 TI - Move over doctor. PMID- 10455700 TI - A new place to call home. PMID- 10455701 TI - What is hay fever? PMID- 10455702 TI - There are so many government initiatives on the health service that managers are drowning in them. PMID- 10455703 TI - Paying for choice in childbirth. PMID- 10455704 TI - Meningitis: time to panic? PMID- 10455705 TI - The aftermath. PMID- 10455706 TI - Brain alert. PMID- 10455707 TI - Credit where credit's due. PMID- 10455708 TI - Face to face. Interview by Alex Mathieson. PMID- 10455709 TI - Ward leaders. Action man. Interview by Jenine Willis. PMID- 10455710 TI - How she learned to live with chronic fatigue syndrome. PMID- 10455711 TI - The Glasgow Coma Scale in clinical practice: a critique. AB - Nurses need to be able to assess a patient's neurological state and record changes in this status over time. One instrument for classifying neurological status is the Glasgow Coma Scale (GCS). Its use is now widespread, although it has not been validated. This article describes the weaknesses of this method and points to an alternative. PMID- 10455712 TI - Improving the quality of care for older people. PMID- 10455714 TI - Feeding time. PMID- 10455713 TI - Nurse prescribing: continuing education and support. PMID- 10455715 TI - Marriage of convenience. PMID- 10455716 TI - Mental health. Addicted to self-affirmation? PMID- 10455717 TI - Mental health. Out with the old. PMID- 10455718 TI - Mental health. Sticks and stones. PMID- 10455719 TI - Demanding and in demand. PMID- 10455720 TI - Pre-operative bowel preparation. PMID- 10455721 TI - Are GPs miffed that at long last promoting and developing nursing is a priority? PMID- 10455722 TI - Toxic toll. PMID- 10455723 TI - Cod and cold turkey. PMID- 10455724 TI - The mouse that roared. PMID- 10455725 TI - What is psoriasis? PMID- 10455726 TI - A TV debate that revealed deep prejudices about mental illness. PMID- 10455727 TI - First things first. PMID- 10455728 TI - Biased approach. PMID- 10455729 TI - Aiding and abetting. PMID- 10455730 TI - Endowment mortgages. PMID- 10455731 TI - Learning by degrees. PMID- 10455732 TI - The wonder of ICU. Interview by Jenine Willis. PMID- 10455733 TI - The gentle touch. PMID- 10455734 TI - A single database that will give you access to all the conditions, organisations and groups you ever needed. PMID- 10455735 TI - How substance use affects people with mental illness. PMID- 10455736 TI - Resolving infant sleep problems. PMID- 10455737 TI - Diagnosis and treatment of ovarian cancer. PMID- 10455738 TI - Taking the measure of patient satisfaction. PMID- 10455739 TI - Constipation-2. Treatment. PMID- 10455741 TI - The house that Matt built. Interview by Robert Munro. PMID- 10455740 TI - The government cannot face the fact that charity and sex education begin at home. PMID- 10455742 TI - Labour pains. PMID- 10455743 TI - What are the working time regulations? PMID- 10455744 TI - Carry off matron. PMID- 10455745 TI - 'Supersister' is the way forward for nurses in need of supportive, creative leadership. PMID- 10455746 TI - Own up to envy: it's best for us all. PMID- 10455747 TI - After dark.... PMID- 10455748 TI - Rescue remedy. PMID- 10455749 TI - All day, and all of the night. PMID- 10455750 TI - Sound support. PMID- 10455751 TI - Boxing should be banned. PMID- 10455752 TI - Primary care. Just what the nurse ordered. PMID- 10455753 TI - Ward leaders. Courage to change. Interview by Jenine Willis. PMID- 10455754 TI - Financial advisers. PMID- 10455755 TI - Mental health. Fellow feelings. PMID- 10455756 TI - A close look at clinical governance reveals a collection of familiar concepts. PMID- 10455757 TI - Sexuality in old age: why it must not be ignored by nurses. PMID- 10455758 TI - Health professionals and the sex education of adolescents. AB - This paper is based on research assessing the provision of appropriate and effective sex education to pupils in the early years of their secondary schooling. It aimed to develop an awareness of their views about health professionals working in schools to help with sex education. The pupils had a range of responses to the work: they liked the openness with which medical personnel tackled the subject and welcomed their medical knowledge and the confidentiality they perceived that went along with that. There were also some profoundly negative reactions: gender had a bearing on the pupils' responses, and boys responded more negatively than girls. Differences in pupils' levels of development and sophistication had an impact on how they received the message. This is an edited version of a paper that will appear in a future issue of NTResearch. PMID- 10455760 TI - Haemolysis of blood samples: what it is and how to avoid it. PMID- 10455761 TI - Latex gloves: an increasing irritation to health care staff. PMID- 10455759 TI - The future for mental health nursing. AB - In February 1998 200 mental health nurses attended the RCN third European Mental Health Nursing Conference in Belfast. This paper reports on a delegate questionnaire concerning issues of resourcing, personal safety, skill mix and commissioning. The findings indicate that mental health nurses remain under threat from physical assault. However, the link between human resourcing, skill mix and threatening behaviour appears not to be one which practising nurses associate with their work area. Not surprisingly, there was a definite perception that purchasers of current services have no real understanding of the needs of service users or providers. PMID- 10455762 TI - Nutrition. Taking the tube. PMID- 10455763 TI - Food intolerance: nutritional consequences of adverse reactions to food. PMID- 10455764 TI - Silent aspiration. PMID- 10455765 TI - A Nightingale for our time. PMID- 10455766 TI - Recognizing, understanding, and responding to familiar responses: the importance of a relationship history for therapeutic effectiveness. AB - PROBLEM: To describe aspects of therapeutic effectiveness, including the importance of an ongoing relationship. METHODS: A phenomenological study (n = 14) involving nurse psychotherapists (n = 6) and clients (n = 8). FINDINGS: Participants recognized dense meanings and familiar, shorthand communications evolving from their relationship history. This ability to read another's responses with sensitivity and attunement contributed to a sense of comfort and openness during therapy sessions. CONCLUSIONS: Students should be taught how a relationship history contributes to sensitivity to nuances of meaning and development of an individualized approach. Reimbursement structures must be developed that acknowledge the situated nature of change and the significance of an ongoing relationship. PMID- 10455767 TI - Male caregivers of mentally ill relatives. AB - TOPIC: To describe the lived experiences of male caregivers of severely mentally ill relatives and their perceptions of this nontraditional role. METHODS: A qualitative study of informal interviews with purposively selected male caregivers (N = 10), using an informal interview guide that addressed psychosocial, physical, financial, and crisis-management categories. FINDINGS: The data yielded three themes of caregiving: expressions of burden, duration and depth of commitment, and role affirmation. CONCLUSIONS: The results can help health professionals develop needed education and support services for men in this emerging role and contribute to family health in the community. PMID- 10455768 TI - Shame and the fear of feeling. AB - TOPIC: Toxic shame and the concomitant fear of feeling are core issues needing to be addressed during group therapy with adult survivors of childhood sexual abuse. PURPOSE: To increase awareness of the toxic shame that survivors experience and to describe the impact of group therapy based on a family systems model. SOURCE: The authors' clinical experiences. CONCLUSIONS: Symptom management, repatterning of cognitive distortions, and the improvement of self-care strategies are identified as crucial aspects of healing shame-based feelings and behaviors. Group treatment offers members the opportunity to cease reenacting family rules and roles that create toxic shame. PMID- 10455769 TI - Cognitive therapy for the suicidal patient: a case study. PMID- 10455770 TI - [Marine accidents. Planning an explanatory booklet for the vacation population]. PMID- 10455771 TI - [The importance of a theory ... the conceptual model by Dorothea E. Orem]. AB - The necessity to care by theoretical thinking and a scientific method is recognized. We have to abandon the task-oriented care model in order to consider a method based on theory to guide our practice. The importance of a Nursing theory depends on its utility in caring and in the disciplinary development. Nurses must learn to be an autonomous profession, based on scientific and theoretical knowledge. Many nursing Scientists have elaborated their own model or theory useful to the Nursing discipline. One of these, is D. Orem's conceptual framework that has defined "self-care" as a human need. In the frame of self care, she has identified the function of Nursing. PMID- 10455772 TI - [Who is afraid of a well prepared nurse? Nursing education: leadership by nurses is urgently needed]. AB - We discuss some problems related to the nursing education and the lack of unitary nursing leadership. The cultural development of the profession is slow and not enough supported. The profession must learn to invest in Nursing education for a real empowerment. We underline some aspects that we think to be important for the future: the revision of the basic nursing curriculum, the teaching position at University, the utility of a post basic nursing degree. In conclusion, some urgent aims for the near future in order to advance the profession, are reported. PMID- 10455773 TI - [Collaboration for community health]. PMID- 10455774 TI - [Quality in drug administration: review of the therapeutic information system during hospitalization]. AB - The mistakes taking place in therapy administration are regarded as indicative elements of a bad work quality. In order to avoid them and to get the best obtainable quality, the Arezzo USL Administration has established a Work Group charged to ascertain the possible errors and to review the different methods enforced inside the informations therapeutic system. An experimentation has been carried out in sixteen hospital wards, with a subsequent control of the results. In all the U.U.O.O. of Arezzo the "Thermography with direct control" system has been adopted, as it has proved to succeed in eliminating a great deal of the possible risks. PMID- 10455775 TI - [Private nursing in public institutions]. AB - In this article the authors review the two sides of the intramoenia private nursing profession: the italian laws and the disciplinary frame of nursing, in order to define bonds and opportunities offered to nursing profession by means of such a topic. PMID- 10455777 TI - [A strategy for nursing research]. AB - The Author reports the conclusions of the 20th meeting of the WENR underlying the importance and the resource of the Council of Europe document on Nursing Research (this document is available in Italian ("Professioni Infermieristiche" 1/98). The action plans for the future as concerns the workgroup activities are also discussed. PMID- 10455778 TI - [The lamp and the fox]. PMID- 10455779 TI - [Humanistic knowledge and scientific research: necessities for nurses]. AB - The way in which humanistic knowledge and scientific research form a binomial inseparable for nurses is pointed out in this article. Notwithstanding the strictness requested by care models, nurses haven't to be turned aside from the quality aspects of an holistic patient care. As the humanistic sciences lead to consider the patient needs as problems to be solved, there are just these problems that stimulate the research work for the purpose of providing the patient with a performance concerning him as a whole. PMID- 10455780 TI - [Employment conditions for graduate nurses in the province of Ravenna from 1994 96]. AB - This article points out the problems that nurses face in order to fit into the working field, together with the present occupational levels and the working area situation. Moreover, the Authors show the particular position of nurses of the three Ravenna's nursing schools graduated in 1994-95-96. PMID- 10455781 TI - [Verification and evaluation of services from the viewpoint of continuing improvement]. AB - Starting from an analysis of the rules of the scenary and context, in which the process of transformation of the health structures in firm is developing, we emphasize how the verification and evaluation of the services represent an indispensable element to improve the quality of the hospital "product", to define the level of adequate quality, and to define the costs of the services relative to the required quality. PMID- 10455782 TI - [From communication with the patient to quality of health services]. AB - Which is the link between the relationship with the patient and the services quality? This work aims to show the existence of this link and its influence on both the quality and the cost of the services given to the patient. PMID- 10455783 TI - [Competence]. AB - In this work different aspects of Nursing Competence are examined. In the first part the terminology is clarified together with its historical and cultural Nursing development. Authors analyse the international literature regarding the topic and the thought of eminent Nursing scholars. In the second part some reflections are given regarding Competence in different fields as: legislation, ethic, education and administration as well as Nursing Research. Conclusions aim to offer the path to demonstrate Competence in order to develop Nursing discipline and profession. PMID- 10455784 TI - [Total quality: priority objective to improve the services of health facilities]. AB - The increasing attention turned to the "total quality" inside health services is pointed out in the article. The development of this theme started in the United States and subsequently reached Europe. In Italy only since a few years we are noticing initiatives aiming both to the improvement of health services quality and to the relationship with clients. The main elements of the total quality management are pointed out: they are addressed to the satisfaction of clients expectations in an environment involving the whole health personnel towards the continuous quality improvement. PMID- 10455785 TI - [Management instruments for nursing care: experiences in a local Milano health facility]. AB - The present article brings into focus the problems rising from the changes required to achieve an efficient use of resources in the health world. How do the nursing services deal with the organizational changes? How to program nurses activities within the nursing services? The article tries to discuss some answers to these problems. PMID- 10455786 TI - [From project to realization. Running of a permanent education course]. PMID- 10455789 TI - [Professionalism]. PMID- 10455787 TI - [Participation in practice: new ways of collaboration in health care]. PMID- 10455791 TI - [Analysis of the activities of ambulatory care nurses and a methodology for the definition of nurses' workload]. AB - Patients need medical and qualified surgery care for different purposes: to begin a diagnostic investigation, to monitor a medical therapy, to execute controls in view of or after a surgical operation, etc. Medical care becomes the heart of patient interest, the direct consequence is a lacking reflection about the purpose of nursery activity in this area and, therefore, on the method to be used to define the staff needed. On this particular subject is based the contribute of this article, considering it of particular interest, being the growing attention on the management of human resource in the health word. PMID- 10455790 TI - [Postoperative pain in an urology department: evaluation by patients and by nurses]. AB - A survey conducted in an urological ward of an Italian hospital, has studied the frequency of pain and altered vital signs connected with pain, in the first 36 postoperative hours. Aim of the study was to locate possible differences between pain assessment performed by patients and nurses. 30 patients operated of urological major surgery have been studied. More altered vital sings are pulse and blood pressure, instead respiration is more altered during the night. Pain is most important in the first 24 hours, particularly at 6 a.m. in the first day after surgery. For patients the most painful operation is radical prostatectomy. Nurses had generally substimated pain; only in the 22% of cases there was agreement with the patients, but with they were agreed that the most painful surgical operation is radical prostatectomy. Vital signs alterations are connected with pain peak only in the 25.36% of cases. PMID- 10455792 TI - [Nurses, woman, breast cancer: integration in a surgical unit. Study of the needs of patients and possible solutions]. AB - The authors report the results of an investigation performed in a surgical teaching department of the University of Padua. A form was given to all women admitted for breast cancer. Questions were related to the most important concerns during the hospital stay. The targets of this investigation were: To evaluate the feelings during the diagnostic work-up and the related procedures, to evaluate the role and the reaction of the patient's family, to define kind and quality of the informations given to the patients from the nurses staff, to report concern about the future, to define quality of nursing associate, ght to detect the most difficult moments of the disease. On the basis of this investigation the authors suggest the options to approach the problems and possible solutions. PMID- 10455793 TI - [The nurse educator: an experience among immigrants from developing countries]. AB - Immigration from developing countries to Italy is a recent fact; however, it is causing different problems, even to health. Among these, tuberculosis holds a position of prominence; it is usually endemic in its original countries, but it finds good condition to become acute again in the discomfort and poverty of home countries. The purpose of the work is to test and confirm the promoting and defensive function of health in nursing profession, thanks to a health education project for immigrants which aims at reducing tuberculosis presence. The instrument used is a lesson about tuberculosis characteristics which the proposal to make the tubercular test, right instrument for a precocious diagnosis. The positive results achieved, show us the right evidence that the nurse is an educator: immigrants have also been offered the possibility to defend their health. PMID- 10455794 TI - [Smoking in a sample of nursing students in Genoa. Reflections on the role of professional educators]. AB - Of all European Union member states, Italy has one of the highest rates of smokers among health care personnel. One important step towards the development of a "prevention-oriented culture" is the raising of the awareness of professional working in health care structures in order to involve them in preventive actions, this making these more effective. Before undertaking measures of this kind, it is imperative to first assess the extent of the smoking habit, even at the local level, so as to target actions as precisely as possible. We herein present the results of a study carried out in two professional nursing schools of the Province of Genoa (I). In all, 341 subjects, aged between 18 and 34, were interviewed. Findings show a high percentage of smokers among both sexes, with 50.0% of males and 40.0% of females claiming to be smokers. These figures are well above national averages (32% of males and 26% of females in the national population). The age at which most interviewees began smoking ranged from 15 to 18 years; however, a rather high percentage of respondents (25.1% for males and 26.1% for females) began after 18 years of age. Our results confirm the importance of school age oriented health education programs, but highlight as well the usefulness of similar awareness initiatives for future professionals. The Italian League for the Fight Against Cancer and the Italian Association of Cancer Nurses propose to vocational training schools a collaborative effort aiming to promote healthy lifestyles among nursing students and to re-assess the role of trainers. PMID- 10455795 TI - [Patient training in peritoneal dialysis: 1. Training in self care, cognitive study]. AB - Authors consider education for dialytic patients a tool for patients' self-care and better quality of their life. In our Country the peritoneal dialysis involve 10% of the total dialytic population where the patient assumes a quite emerging role. The educational process is called training and the responsibility of the nurse is primarily important due to the trust and the consequential nurse-patient relationship. The article is divided in two parts: in the first, we explore the importance of the training as teaching methodology and in the second, we present the results of a national survey to better understand the role of the nurse and the methodology utilised in different Centers. PMID- 10455797 TI - [Experiences in a psychiatric nursing service]. PMID- 10455798 TI - [A new view of critical care]. PMID- 10455801 TI - [A stimulating initiative of the International Council of Nurses: the opening of a new Internet site]. PMID- 10455799 TI - [National congress of the CNAI in Pesaro. Autonomy of the nursing profession: state of the art]. PMID- 10455802 TI - [XVI special national congress of CNAI. A challenge for the future of CNAI at 50 years of membership in the International Council of Nurses]. PMID- 10455803 TI - [Autonomy of the nursing profession: state of the art]. AB - This article points out the foundation elements of the nursing education, the way covered till now, the status of the art and what has to be achieved in order to attain Nursing Degree objectives. The different aspects concerning the curricula organization and the model of care are examined. The request of getting nursing teaching and nursing chairs for nurses are an aim not yet met, but they are the basis objectives of the professional engagement. Another important subject concerns the leadership that will allow nurses to be administrators and managers in every respect, also from a juridical point of view, and to express their professionality in a really autonomous way. PMID- 10455804 TI - [How to overcome problems of a nursing education towards professional autonomy]. AB - This article, through an historical report of the legislation concerning nursing education, shows "schizophrenia role" that brings to a confusion. The professional authonomy, by turns denied or asserted, has always been for us the "litmus paper" indicating a series of difficulties interfering with the professional development. We believe that only in a "global education" (D.V.- Nursing Degree) we can find the natural solution of the difficulties met in Italy by a nursing profession that is young from an historial point of view, but is aware of its education needs. PMID- 10455805 TI - [An active way for professional autonomy]. AB - Authonomy, competence, specificity. What is nowadays the situation of the professional practice authonomy? To answer the question, the author presents an outline of the present condition in this field. Different possible developments and solutions are shown, starting from some interventions at a personal level to demonstrate nursing specificity to go up to superior levels, belonging to the professional organs. PMID- 10455806 TI - [Degrees for nursing leaders. Possible strategies]. AB - The three conditions existing for the establishment of a Nursing Science Degree, analysed in this article, are pointed out as the ones on which relies the urgent request of getting a Nursing Science degree, put forward by the nursing profession. They are: 1) The existence of a Nursing Science 2) Nursing care is now carried out in systems with a complex organization 3) A Nursing degree is requested by the Health Institutions to be admitted to leadership. This aim has not yet been achieved only owing to the political, institutional and professional Italian turbulences. PMID- 10455807 TI - [Economic and social recognition: where are we?]. AB - The Author analyses the contemporary Nursing working situation referring it to the economical point of view. A discussion is reported on the new working contract and on the future of professionals looking at the economical and jiuridical classification. PMID- 10455808 TI - [A standard type of nurse. An Italian experience]. AB - There is a great deal of rhetoric surrounding the notion of a standard type of nurse. With claims of the globalisation of the nursing profession along with many other professions in the late twentieth century we risk oversimplifying the possibility of an actualisation of the "standard nurse". The following article looks briefly at the conditions and issues that confront a group of nurses working in a Rome intensive care unit. It argues that many of the difficulties they face as nurses arise from the incompatibility of the notion of a standard type of global nurse with their workplace conditions, professional support mechanism and various cultural and management based notions chich strongly influence their held identity. PMID- 10455809 TI - [Nature and scope of the examination and review of the quality of care]. AB - The quality is a dynamic and multidirectional phenomenon that depends on many factors. It changes in the time in relation to the variation of the cultural and social behaviours, of the scientific and technologic knowledges and to the change of the natural resources. PMID- 10455810 TI - [Psychiatric nursing yesterday and today]. AB - Psychiatry, as we know it today, and chiefly the role the nurse plays inside the different psychiatric services on the territory, are a result of the psychiatry cultural growth, during the centuries, among many contradictions. Psychiatric support and the nurse's image have always been bound, till recently, to the mental hospital as an institution. After the 180/78 law, the responsibility of the psychiatric support has passed to the territory and, with this change, the way of conceiving the psychiatry and the supporting approach to the patient have changed too. The professional man, who wants to offer a scientific support and to raise the quality of his service, has to carry out a scientific methodology using a theoric model as landmark and suitable operating tools. PMID- 10455811 TI - [Nursing care of psychiatric emergencies]. AB - Starting from an analysis of the different psychiatrical emergencies, and devoting the attention to the more frequent events, we analyse the guide lines the nurse must conform to, so that he can satisfy psychiatrical patient's demands, and provide him with a specific relief, in order to get better quality of his life, and at the same time offering a substantial and high quality assistance. PMID- 10455812 TI - [Aspects of total parenteral nutrition]. AB - Parenteral Nutrition has considerably increased the responsiveness to medical or surgical treatment of a patient. All the nourishing infused substances are in an elementary state, ready for a direct utilization by the cells. They offer a caloric and proteinous contribution which is necessary to the metabolic demands of an organism that grows in case of fewer, traumas, scalds, in post surgical time, or in hypermetabolic pathologies. The physician, nurse, and dietician decide the necessity to Total Parenteral Nutrition, valuing the patient's parameters such as: loss of weight, azotic budget, percentage of loss of muscular and lean mass, the patient's inability to tolerate food through gastroenteric way. A careful valuation and a scrupulous treatment by physician and nurse can reduce the risk of complications during parenteral feeding. PMID- 10455813 TI - [Prevention and treatment of decubitus ulcers: an experience]. AB - We analyze how pressure sores influence hospital stay. In the first part, the risk factors are pointed out; we define the aims and nursing actions to prevent the phenomena. The Authors present their assessment kardex along with the protocols related to different stages of the sore. At end, a tentative of efficacy's evaluation is reported. PMID- 10455817 TI - [The principal stages of activity of the International Council of Nurses for the nursing profession and for health]. PMID- 10455818 TI - Overcome terminal seriousness: let go, laugh, and lighten up! AB - This article examines physiological and psychological benefits of laughter and humor. These are essential tools that can successfully combat the stress and pressure of the perioperative environment. Used in tandem with "life balancing skills," they can enrich the work arena and help nurses rekindle passion and meaning in their lives. PMID- 10455819 TI - Incorporating fun into the business of serious work: the use of humor in group process. AB - Incorporating formal humor and creative play into group process improves the outcome of serious work. This article discusses the use of humor and play in a traditionally serious setting. On-the-job playfulness improves communication, creativity, problem solving, and team building, thereby increasing productivity and satisfaction. PMID- 10455820 TI - Keeping them in stitches: humor in perioperative education. AB - Humor is one of the most effective teaching strategies available to perioperative nurse educators. Humor can be used to teach nursing students, surgical staff, and patients. This article describes the incorporation of humor into perioperative education and presents examples of specific humorous teaching strategies. PMID- 10455821 TI - Humor as a perioperative nursing management tool. AB - Humor is a powerful tool for accomplishing management goals. This article offers suggestions for using humor to relieve stress, cultivate teamwork, buy attention, improve morale, increase productivity, foster communication, enhance problem solving, and create a positive work culture in the operating room. PMID- 10455822 TI - Constipation among operating room nurses: flatulence as evidence. AB - Even perioperative nurse researchers have a sense of humor. This article is a parody of a research study proposal in which the investigator aims to study a problem common to perioperative nurses during long procedures. PMID- 10455823 TI - You had to be there: true stories of humor in the OR. AB - This article is a collection of true, funny stories of life in the operating room, submitted by subscribers to a perioperative listserv. Each story illustrates one or more of the three building blocks of humor: surprise, tension, and relationships. PMID- 10455824 TI - Pre-op instructions as our patients hear them. PMID- 10455825 TI - Laws of the OR. PMID- 10455826 TI - Clinical Governance--what is it all about? PMID- 10455827 TI - A comparison of the effects of dopexamine and prostacyclin in systemic inflammatory response syndrome. AB - Forty critically ill patients fulfilling the definitions of systemic inflammatory response syndrome were enrolled in a double-blind cross-over interventional study assessing the effects of dopexamine and prostacyclin on splanchnic perfusion, cardiovascular function and oxygen flux. The cross-over design involved either dopexamine (1.25 micrograms.kg-1.min-1) or prostacyclin (0.5 ng.kg-1.min-1) being infused for 6 h followed by a 12-h washout period prior to an identical infusion sequence using the other agent. Preliminary analysis revealed a significant period effect, so data from the second infusion period were excluded from further analysis. Dopexamine caused a significant increase in heart rate (116 vs. 106 beat.min-1), and urine output (103 vs. 69 ml.h-1). Dopexamine produced a significant increase in oxygen delivery (infusion 548 ml O2.min-1.m-2; no infusion 492 ml O2.min-1.m-2) while prostacyclin caused a decrease (infusion 460 ml O2.min-1.m-2; no infusion 547 ml O2.min-1.m-2). The results indicate that dopexamine improves oxygen delivery and urine output more effectively than prostacyclin. PMID- 10455828 TI - The effect of oxygen on propofol-induced inhibition of microsomal cytochrome P450 3A4. AB - We have shown previously that both hypoxia and propofol may inhibit the metabolism of midazolam. We now wished to see whether there was any additive or synergistic effect when they occurred together. Microsomes were incubated with 20 microns midazolam for 60 min, and propofol 0, 50, 100 or 1000 microM was added. Incubates were further subdivided so that the environment contained 0, 10, 21 or 70% oxygen. The results confirmed our earlier study showing that propofol only had a significant inhibitory effect at a concentration greater than that seen clinically (1000 microM). Anoxia was the only environment in which significant depression of the metabolism of midazolam occurred at all concentrations of propofol. This reduced it to almost zero. Post hoc analysis of the data showed that, with the greatest concentration of propofol (1000 microM), there was increasing inhibition of metabolism of midazolam with increases of oxygen from 10 to 70%. PMID- 10455829 TI - Ventilatory pattern and associated episodic hypoxaemia in the late postoperative period in the general surgical ward. AB - Episodic oxygen desaturation is frequent in the late postoperative period and seems most pronounced on the second and third postoperative nights. However, the ventilatory pattern has not been described systematically during this period. We studied the ventilatory pattern and associated arterial oxygenation using the Edentrace II equipment (impedance pneumography and pulse oximetry) on the second and third postoperative nights in 28 patients undergoing major abdominal surgery. Ventilatory disturbances were common and included periods of hypopnoea, and obstructive, central and mixed apnoeas. Overall, the median (range) respiratory disturbance index (apnoeas + hypopnoeas per h) was 12 (0-121), with the patients spending 6% (0-65%) of the night in some kind of ventilatory disturbance. It was not possible from pre-operative snoring habits to predict patients who developed postoperative ventilatory disturbances. Overall, 23% (0-100) of the hypopnoeas and 7% (0-100) of the apnoeas were associated with episodic hypoxaemia. In conclusion, ventilatory disturbances were common in the late postoperative period in the general surgical ward and often associated with episodes of oxygen desaturation. PMID- 10455831 TI - Xenon: recent developments. PMID- 10455830 TI - Eating in labour. A randomised controlled trial assessing the risks and benefits. AB - The aim of this study was to determine whether permitting women in labour to eat a light diet would: (i) alter their metabolic profile, (ii) influence the outcome of labour, and (iii) increase residual gastric volume and consequent risk of pulmonary aspiration. Women were randomised to receive either a light diet (eating group, n = 48) or water only (starved group, n = 46) during labour. The light diet prevented the rise in plasma beta-hydroxybutyrate (p = 2.3 x 10(-5)) and nonesterified fatty acids (p = 9.3 x 10(-7)) seen in the starved group. Plasma glucose (p = 0.003) and insulin (p = 0.017) rose in the eating group but there was no difference in plasma lactate (p = 0.167) between the groups. There were no differences between the groups with respect to duration of first or second stage of labour, oxytocin requirements, mode of delivery, Apgar scores or umbilical artery and venous blood samples. Relative gastric volumes estimated by ultrasound measurement of gastric antral cross-sectional area were larger (p = 0.001) in the eating group. This was supported by the observation that those from this group who vomited, vomited significantly larger volumes than those in the starved group (p = 0.001). We conclude that eating in labour prevents the development of ketosis but significantly increases residual gastric volume. PMID- 10455832 TI - Evaluation of the SCOTI device for confirming blind nasal intubation. AB - The sonomatic confirmation of tracheal intubation (SCOTI) is a new device used to confirm the correct placement of tracheal tubes. It utilises a sonic technique for recognition of a resonating frequency for detection of tracheal intubation. We compared its predictive value with that of the clinical auscultatory method and a capnograph to confirm 132 blind nasal intubations using three different tracheal tubes [red rubber (n = 82), polyvinyl chloride (n = 33) and RAE preformed nasal (n = 17)]. SCOTI correctly identified 70.8% of intubations and chest auscultation did so 99.2% of times. All results were confirmed using a capnograph. The SCOTI device gave a false-negative value in 37 patients (28%) and a false-positive result in two patients (1.5%). The response time for confirming intubations was 2.5 (1.5) s for the SCOTI, 4.1 (1.1) s for a capnograph and 40 (9.4) s for the auscultatory method. The erroneous results shown by the SCOTI device were highest when polyvinyl chloride tubes with a Murphy's eye were used for intubation. This study shows that this device is not very useful for ascertaining the correct placement of tracheal tubes after blind nasal intubation. PMID- 10455833 TI - Spinal haematoma following epidural anaesthesia in a patient with eclampsia. AB - A patient with a twin pregnancy required a Caesarean section for severe pre eclampsia. Her platelet count was 71 x 10(9).l-1. Epidural anaesthesia was performed after platelet transfusion. A spinal epidural haematoma was diagnosed postoperatively. A generalised tonic-clonic seizure sparing the lower limbs enabled early diagnosis to be made. The patient recovered with no permanent neurological damage after laminectomy and clot removal. The risks and benefits of regional techniques require careful consideration, and postoperative monitoring for recovery of neural blockade is essential. PMID- 10455834 TI - Spinal anaesthesia for caesarean section in a patient with systemic sclerosis. AB - We describe the management of a diabetic primigravid woman with systemic sclerosis and thrombocytopaenia who required Caesarean section for pre-eclampsia. This was performed successfully under spinal anaesthesia. PMID- 10455835 TI - Use of the flexible intubating fibrescope for tracheal re-intubation in a patient with difficult extubation. AB - Difficulty in tracheal extubation was experienced in a patient following major maxillofacial surgery for reconstruction of the maxilla using bone grafts and a microvascular free flap. With the aid of an intubating flexible fibrescope, the cause of the difficulty was identified as a stitch accidentally transfixed through the tracheal tube. Tracheal re-intubation was required to facilitate surgical exploration to remove the stitch and the proximal end of the tube. The tip of an Olympus LF-2 intubating fibrescope was successfully negotiated in the trachea alongside the original tube with its cuff deflated. This allowed safe and speedy railroading of a new tube immediately after the distal end of the original tube was removed. PMID- 10455836 TI - Cor pulmonale presenting in a patient with congenital kyphoscoliosis following intercontinental air travel. AB - We present the case of a 59-year-old man with congenital kyphoscoliosis who developed cor pulmonale for the first time following intercontinental air travel. Prolonged exposure to the low partial pressure of oxygen in the cabin of the aircraft led to pulmonary hypertension and right heart failure. The case highlights the potential for long-haul air travel to cause decompensation in patients with thoracic deformity and apparently stable cardiorespiratory function. It also emphasises the need for patients and their medical attendants to carefully consider the potential health implications of the hypoxic atmosphere in pressurised aircraft. PMID- 10455837 TI - A case of ventricular fibrillation in the prone position during back stabilisation surgery in a boy with Duchenne's muscular dystrophy. AB - A 15-year-old boy with Duchenne's muscular dystrophy experienced prolonged cardiac arrest whilst in the prone position for spinal surgery. He was successfully resuscitated without apparent neurological sequelae by internal cardiac massage via a thoracotomy and external and internal direct current cardioversion. Recommendations are suggested for the pre- and peroperative management of such cases. PMID- 10455838 TI - Epidural placement in a patient with undiagnosed acquired haemophilia from factor VIII inhibitor. AB - We report a case of acquired haemophilia that presented as continuous bleeding from an epidural insertion site in a previously asymptomatic parturient. Peripartum anaesthetic management of this rare medical emergency has not previously been reported and the difficulties associated with its treatment are discussed. PMID- 10455839 TI - An audit of airway problems in the recovery room. AB - It has recently been suggested that recovery rooms should have dedicated anaesthetic cover during working hours to deal with serious life-threatening problems. This audit was undertaken to determine the incidence and severity of airway problems that occurred in the recovery room at a District General Hospital. One thousand consecutive patients who received a general anaesthetic were assessed. If an airway problem was identified, the patient's notes were examined to document the type of surgery and any predisposing factors which may have contributed. The incidence of airway problems in this study was found to be 2.8%, which was in agreement with previous studies and appears too low to warrant a full-time anaesthetic presence. PMID- 10455840 TI - Rapid sequence induction: a questionnaire survey of its routine conduct and continued management during a failed intubation. AB - Two hundred and ten obstetric anaesthetists completed a questionnaire assessing how they would perform a rapid sequence induction of anaesthesia for a Caesarean section and their continued management during a failed tracheal intubation. The survey revealed considerable variation in the timing and application of cricoid pressure, the choice and dose of drugs used and the timing of their administration. The management of a difficult intubation also varied. This variability was independent of both grade of anaesthetist and frequency of practice. There appear to be at least two distinct techniques in current practice, characterised by 'fast' or 'slow' rapid sequence induction. Rapid sequence induction is clearly not a standard technique and debate is necessary to clarify the risks and benefits of its components. In particular, the rapidity of the technique and the application of cricoid pressure may contribute to the increased incidence of difficult tracheal intubation in obstetric anaesthesia. PMID- 10455841 TI - The analgesic efficacy of tramadol versus ketorolac in day-case laparoscopic sterilisation. AB - We conducted a prospective, randomised, double-blind study to compare the analgesic efficacy of intravenous tramadol 1.5 mg.kg-1 and ketorolac 10 mg in 60 ASA grade 1 and 2 patients scheduled to undergo day-case laparoscopic sterilisation by application of Filshie clips. Patients who received tramadol had significantly less postoperative pain in the recovery room (p = 0.007) and at discharge from the day-surgery unit (p = 0.03), and they required rescue analgesia with morphine less often (p = 0.02) than patients who received ketorolac. No difference in either the incidence or severity of nausea and vomiting was observed between the two groups. Both analgesic drugs were well tolerated at the doses given in the study, although dry mouth was significantly more common after the administration of tramadol (p = 0.009). Three patients in the tramadol group and five in the ketorolac group required overnight admission due to pain or nausea and vomiting. PMID- 10455842 TI - Why do patients feel positive about patient-controlled analgesia? AB - We studied 200 patients to identify the aspects of their experience of patient controlled analgesia (PCA) that made them feel 'extremely positive' about this technique. After PCA had been withdrawn, patients completed a questionnaire which included the following topics: pre-operative information, pain relief, the degree of control that PCA afforded the patient, side-effects and safety. Multiple regression analysis identified three factors of their experience which were associated uniquely with feeling 'extremely positive' about PCA: having better pain relief, not worrying about 'giving oneself too much drug' and not experiencing feeling 'peculiar in the head'. Control over pain relief, although highly correlated with feeling 'extremely positive' about PCA, was unimportant when these variables were controlled. Because of the well-recognised difficulties in measuring satisfaction with analgesic regimens, we suggest that a satisfaction score based on these variables would be a significant advance on existing methods. PMID- 10455843 TI - Intravenous buscopan for analgesia following laparoscopic sterilisation. AB - Following reports that tubal smooth muscle spasm may contribute to pelvic pain following laparoscopic sterilisation, we studied the effect of buscopan (an anticholinergic agent used to relieve smooth muscle spasm) on 45 patients undergoing general anaesthesia for day-case laparoscopic sterilisation. Patients were randomly allocated to receive either buscopan 20 mg or saline placebo after induction of anaesthesia. There were no significant differences in pain scores or postoperative analgesic requirements between the two groups. We conclude that intravenous buscopan confers no benefit in day-case laparoscopic sterilisation. PMID- 10455844 TI - Paediatric intensive care--the way ahead. PMID- 10455845 TI - Peri-operative steroid supplementation. PMID- 10455846 TI - Nitroglycerin for intra-operative bronchospasm. PMID- 10455847 TI - Advantages of the combined spinal epidural (CSE) technique. PMID- 10455848 TI - Advantages of the combined spinal epidural (CSE) technique. PMID- 10455849 TI - Safety and technique with pencil-point subarachnoid needles. PMID- 10455850 TI - Helicobacter pylori: an occupational hazard for anaesthetists. PMID- 10455851 TI - Pregnancy-induced cholinesterase deficiency. PMID- 10455852 TI - Suxamethonium and intra-ocular pressure. PMID- 10455853 TI - Lead time bias and standardised mortality ratios in intensive care patients. PMID- 10455854 TI - Internal jugular vein cannulation confused with carotid artery cannulation. PMID- 10455855 TI - Avoid internal jugular vein transfixion. PMID- 10455856 TI - The intubating laryngeal mask airway in failed and difficult intubation. PMID- 10455857 TI - The cuffed oropharyngeal airway. PMID- 10455858 TI - Conscious sedation for fibreoptic intubation. PMID- 10455859 TI - High-frequency oscillation via a laryngeal mask airway. PMID- 10455860 TI - Pharyngeal wall perforation--an unusual complication of blind intubation with a gum elastic bougie. PMID- 10455862 TI - A potential hazard with TracheoSoft PERC tracheostomy tubes. PMID- 10455861 TI - A serious breathing system fault. PMID- 10455863 TI - High airway pressures with sticking one-way valves in a circle system. PMID- 10455864 TI - A successful introduction to central venous cannulation? PMID- 10455865 TI - Hoarse voice and fine-bore nasogastric tubes. PMID- 10455866 TI - An alternative intravenous route. PMID- 10455867 TI - Intubation through a hedge. PMID- 10455868 TI - The bearded airway. PMID- 10455869 TI - All the fun of the fair. PMID- 10455870 TI - Caring for stockings. PMID- 10455871 TI - SUS mentorship in Y2K--passion, leadership, perspective. PMID- 10455872 TI - Immune selection after antigen-specific immunotherapy of melanoma. AB - BACKGROUND: Melanoma antigen (MA)-specific vaccination strongly enhances antitumor reactivity in vivo and is capable of producing strong cytotoxic T lymphocyte responses in vitro. Furthermore, specific human leukocyte antigen restricted T cell activation is hypothesized to occur in response to peptide based immunotherapy, which may lead to the preferential killing of tumor cells bearing the relevant MA. The development of melanoma antigen-loss variants may subsequently occur in vivo. METHODS: Analysis of 532 melanoma lesions from 204 patients was performed on fine-needle aspiration biopsy specimens. Lesions were graded for the expression of the MAs gp100 and MART-1 with use of immunocytochemistry. A total of 351 melanoma lesions were divided into cohorts on the basis of the treatment received. The pretreatment group (n = 175) consisted of lesions obtained before any form of gp100 immunotherapy, with the posttreatment group (n = 176) consisting of lesions obtained after vaccination with a modified gp100 epitope, gp209-2M +/- interleukin 2 (IL-2). RESULTS: The percentage of lesions not expressing the gp100 antigen is greater than the percentage not expressing MART-1 (26% vs 14%). The frequency of lesions with high expression (> 75%) of gp100 significantly decreased with therapy (47% vs 34%) and conversely negative lesions increased (18% vs 29%). Treatment of lesions with peptide alone (no IL-2) revealed a significant decrease in gp100 expression (47% vs 32%), enhanced with the addition of IL-2 to therapy (47% vs 35%). The expression of MART-1 remained essentially unchanged unless IL-2 was added (54% vs 54%, MART-1 peptide alone, 54% vs 43%, MART-1 peptide + IL-2). Of 94 patients (181 lesions) assessed for gp100 expression before treatment, 10 patients responded to therapy. Pretreatment lesions in responding patients expressed some level of gp100 in all cases compared with 27% of nonresponding lesions, which were negative for gp100 expression. CONCLUSIONS. This study indirectly demonstrates that vaccination with an MA-derived peptide can result in immune selection in vivo. Furthermore, it provides strong immunologic evidence for the specificity of MA vaccines and to the relevance of MA expression in predicting the response to vaccination. PMID- 10455873 TI - Alpha beta TCR+ T cells play a nonredundant role in the rejection of heart allografts in mice. AB - BACKGROUND: Although the transplantation of solid organs and cellular grafts is a clinical routine, the morbidity and mortality associated with immunosuppression is significant. This could be avoided by the induction of donor-specific tolerance. To develop targeted antirejection strategies and regimens to induce donor-specific tolerance, cell populations in the recipient-mediating rejection of solid organ and cellular grafts must be defined. In this study we examined the role of alpha beta-TCR+ cells in the rejection of allogeneic heart grafts, by use of knockout (KO) mice deficient in the production of alpha beta-TCR+ T cells. METHODS: C57BL/6-TcrbtmlMom (alpha beta-KO) and C57BL6/J (B6) recipient mice were transplanted with B10.BR/SgSnJ (B10.BR) or BALB/c heart allografts. Animals also received bone marrow from normal B10.BR donors, followed by donor-specific or third-party heart transplants. RESULTS: Naive B6 control mice rejected B10.BR and BALB/c grafts within 16 days. In striking contrast, B10.BR and BALB/c heart allografts were indefinitely accepted in unmanipulated alpha beta-KO mice. The immune responsiveness was restored after bone marrow transplantation from normal donors. After bone marrow transplantation major histocompatibility-disparate BALB/c third-party heart grafts were rejected, whereas donor-specific grafts were still accepted. CONCLUSIONS: alpha beta-TCR+ T cells play a nonredundant role in the rejection of heart allografts in mice. Bone marrow chimerism is associated with donor-specific transplantation tolerance. PMID- 10455874 TI - External support modulates G protein expression and receptor coupling in experimental vein grafts. AB - BACKGROUND: Intimal hyperplasia remains the leading cause of vein graft failure. Various external stenting devices have been shown to reduce the development of intimal hyperplasia in vein grafts. Mitogenic and mechanotransduction signals are known to be mediated by G protein-coupled receptors. Therefore in this study we examined the alterations in G protein expression and receptor coupling in vein grafts stented with external tube support. METHODS: Thirty New Zealand White male rabbits had a right carotid interposition bypass graft with use of the ipsilateral jugular vein. Fifteen animals received external support and 15 were controls. In a subset the animals either had removal of the external support or a sham-control neck exploration at 14 days after the initial implantation (n = 5 per group). RESULTS: External support reduced G alpha i3 proteins by 30% in vein grafts without changes in G alpha s by Western blot. Vein grafts with external support were significantly less sensitive to pertussis toxin inactivation than controls were in response to both norepinephrine and serotonin. A 24% decrease in intimal thickness was maintained after withdrawal of the initial external support. CONCLUSIONS: The placement of an external support is associated with alternations in G protein expression and receptor coupling function in vein grafts. The results of this study suggest that the development of vein graft intimal hyperplasia may involve G protein-mediated events. PMID- 10455875 TI - Liposomal delivery of heat-shock protein 72 into the heart prevents endotoxin induced myocardial contractile dysfunction. AB - BACKGROUND: The purposes of this study were to (1) determine whether functional heat-shock protein 72 (HSP-72) may be delivered into the heart, (2) determine whether HSP-72 itself is protective against endotoxin (lipopolysaccharide [LPS] induced cardiodepression, and (3) compare relative protection and time courses required for protection for thermally induced HSP-72 versus liposomally introduced HSP-72. METHODS: HSP-72 was introduced (liposomal HSP-72) or induced (heat shock, 42 degrees C x 15 minutes, 24 hours before) in rat heart before LPS administration (0.5 mg/kg intraperitoneal or ex vivo coronary infusion). Western blot analysis for HSP-72 was used to confirm its expression. Left ventricular developed pressure (Langendorff) was used as an index of cardiac function. RESULTS: Direct intracoronary perfusion of liposomal HSP-72 delivered functioning HSP-72 into the myocardium. LPS induced cardiodepression; however, heat shock pretreatment abolished LPS-induced contractile dysfunction. A direct connection was found between HSP-72 and protection derived from liposomal transfer experiments that similarly reduced LPS-induced cardiodepression. CONCLUSIONS: (1) HSP-72 prevents LPS-induced myocardial contractile dysfunction, (2) liposomal transfer of HSP-72 into the myocardium provides the first direct mechanistic connection between myocardial HSP-72 and protection against LPS, (3) HSP-72 induction requires 24 hours and liposomal transfer of HSP-72 requires 90 minutes, and (4) HSP-72 may offer a clinically acceptable means of protecting the heart. PMID- 10455876 TI - Inhibition of cytokine-induced nitric oxide synthase expression by gene transfer of adenoviral I kappa B alpha. AB - BACKGROUND: Nitric oxide is overexpressed in nearly every organ during sepsis and it has profound biologic effects. Previously, we showed that maximal inducible nitric oxide synthase (iNOS) expression is up-regulated by a combination of cytokines and that this effect is mediated by the transcription factor NF-kappa B. Therefore the purpose of this study was to establish whether gene transfer of the inhibitory molecule I kappa B would result in the abrogation of cytokine induced iNOS expression. METHODS: Cultured hepatocytes were infected with an adenoviral vector containing the I kappa B alpha gene (Ad5I kappa B) and after an 18-hour recovery period were stimulated with the cytokine mixture of tumor necrosis factor-alpha (500 U/mL) plus interleukin 1 beta (200 U/mL) plus interferon gamma (100 U/mL). RESULTS: As expected, cytokine mixture induced significant hepatocyte nitrite (NO2-) and iNOS messenger RNA production. Cells infected with the I kappa B alpha gene showed a dose-dependent decrease in NO2- and iNOS messenger RNA levels. Western blot analysis showed a marked decrease in iNOS protein levels in the presence of Ad5I kappa B alpha. Gel shift assays of nuclear extracts demonstrated that Ad5I kappa B alpha decreased the cytokine induced DNA binding activity for NF kappa B. CONCLUSIONS: NF kappa B is an important regulator of cytokine-induced NO expression. These results identify a novel therapeutic approach where gene transfer of the inhibitory molecule I kappa B alpha can be used to down-regulate cytokine-induced iNOS expression as well as other NF kappa B-dependent genes that are up-regulated during the inflammatory response. PMID- 10455877 TI - Mitochondrial DNA damage and altered membrane potential (delta psi) in pancreatic acinar cells induced by reactive oxygen species. AB - BACKGROUND: Reactive oxygen species (ROS) have been implicated in the induction of acute pancreatitis. Mitochondria possess a distinct genome (mtDNA) that is more susceptible to ROS-induced damage than nuclear DNA (nDNA). The purpose of our study was to determine the effect of ROS on mitochondrial function and membrane potential (delta psi mt), to identify signal transduction mechanisms activated by ROS, and to quantify damage to mtDNA in an in vitro pancreatitis model. METHODS: Pancreatic acinar cells, AR4-2J, were treated with saline solution (control) or hydrogen peroxide (H2O2), a representative ROS. Mitochondrial function was assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay; to determine delta psi mt, rhodamine-123 uptake was measured. Intracellular calcium levels and c-Jun N-terminal kinase activity was determined; gel mobility shift assays were performed to assess induction of the transcription factor NF-kappa B. To quantitate DNA damage, a novel polymerase chain reaction-based procedure was performed. RESULTS: Mitochondrial function and delta psi mt were significantly decreased with oxidative damage. H2O2 treatment resulted in increased intracellular calcium levels, activation of c-Jun N-terminal kinase, and induction of NF-kappa B DNA binding. Treatment of AR4-2J cells with H2O2 resulted in selective mtDNA damage; nDNA was not affected. CONCLUSIONS: Our data demonstrate that pancreatic mtDNA is more susceptible to oxidative damage than nDNA; this damage is associated with decreases in mitochondrial function and delta psi mt and activation of downstream signal transduction pathways. Mitochondrial damage mediated by ROS may play a central role in pancreatic cell injury associated with acute pancreatitis. PMID- 10455878 TI - Timing of sentinel lymph node mapping after lymphoscintigraphy. AB - BACKGROUND: Sentinel lymph node (SLN) mapping is an effective technique for staging patients with melanoma. In an attempt to avoid reinjection of radiolabeled colloid and facilitate SLN mapping at the time of surgery, we examined whether residual radioactivity from preoperative lymphoscintigraphy could be used to accurately identify SLNs during surgery 18 to 24 hours later. METHODS: Forty-six patients with newly diagnosed melanoma underwent injection of 0.22-micron filtered technetium 99m-labeled sulfur colloid followed by lymphoscintigraphy. Patients returned the next day for SLN biopsy with Isosulfan blue dye and the hand-held gamma-probe to identify SLNs. Thirty of 46 patients underwent repeat imaging before operation. No patient had reinjection of radiocolloid. RESULTS: Ninety-five SLNs were identified on initial lymphoscintigraphy, and repeat imaging on the day of surgery confirmed all SLNs previously identified. A total of 122 SLNs (2.65 per patient) were resected from 58 basins. Eighty-four (69%) of 122 SLNs stained blue, and 118 (97%) of 122 SLNs had in vivo gamma-counts greater than 4 times background. Microscopic metastases were present in 13 (10.7%) of 122 SLNs in 12 (26.1%) of 46 patients. There have been no recurrences over a mean follow-up time of 320 days. CONCLUSIONS: Intraoperative gamma-probe detection combined with blue dye injection is highly effective in identifying SLNs 18 to 24 hours after injection of 0.22-micron filtered 99mTc-sulfur colloid. Reinjection of radiocolloid is not required. This technique avoids radiopharmaceutical administration in the operating room, minimizes radiation exposure, and increases scheduling flexibility. PMID- 10455879 TI - Direct genetic testing for Smad4 mutations in patients at risk for juvenile polyposis. AB - BACKGROUND: The identification of germline mutations in juvenile polyposis (JP) families has made presymptomatic genetic testing possible. In this study we report the results of genetic testing in two large JP families and develop an algorithm for the clinical management of these patients. METHODS: DNA was extracted from 55 members of 2 JP kindreds, and the Smad4 mutations in the germline were determined by direct sequencing. All family members were then tested for mutations with use of single-strand conformational polymorphism analysis and were invited for genetic counseling. RESULTS: All 18 affected members of both kindreds had a 4-bp deletion in exon 9 of the Smad4 gene. In 30 patients at risk for JP, 17 had previously had negative endoscopic screening results and 13 had never been screened. Five patients at risk had inherited germline Smad4 mutations. Two carriers have had hematochezia but have not been screened, whereas 3 were asymptomatic. The mean age of carriers was 29.8 years (range 9.1-49.5 years), whereas that of noncarriers was 41.0 years (range 8.1 76.5 years). CONCLUSIONS: Compliance has been a problem with endoscopic screening for JP. With genetic testing non-carriers may no longer require frequent screening endoscopy, whereas gene carriers can be targeted for close endoscopic surveillance and early intervention to prevent the development of gastrointestinal cancers. Direct genetic testing significantly improves the presymptomatic diagnosis of gene carriers in JP families with Smad4 mutations. PMID- 10455880 TI - Adenovirus-mediated gene transfer to the peritoneum and hepatic parenchyma of fetal mice in utero. AB - BACKGROUND: The development of effective gene transfer in utero will provide alternative approaches to the treatment of genetic disorders. For many disorders, the fetal liver and peritoneum are important target tissues. Our goals were to compare the tissue sites and duration of transferred gene expression after intraperitoneal (i.p.) or intrahepatic adenoviral-mediated gene transfer in utero in the developing murine fetus. METHODS: Day 15 CD-1 fetuses were injected intrahepatically or intraperitoneally with recombinant adenoviruses containing the luciferase or beta-galactosidase reporter gene. Tissue levels of luciferase were quantitated, or tissues were examined for X-gal staining. RESULTS: Luciferase expression was observed in multiple fetal tissues (including brain, intestine, liver, and lung) and persisted up to 32 days after intrahepatic delivery. Significant hepatic tropism was demonstrated. CONCLUSIONS: Intrahepatic and intraperitoneal injection in utero results in transduction of multiple tissues in the developing murine fetus. Transuterine injection of fetal mice via intrahepatic and intraperitoneal routes provides a valuable model for assessing the efficacy of gene delivery vectors in the prenatal treatment of genetic disorders. These studies demonstrate that hepatic and intraperitoneal gene transfer to the developing murine fetus is feasible and may provide therapeutic levels of proteins during fetal development. PMID- 10455881 TI - Relationship between hospital volume and late survival after pancreaticoduodenectomy. AB - BACKGROUND: Several studies have reported lower perioperative mortality rates with pancreaticoduodenectomy at high-volume hospitals than at low-volume hospitals. We sought to determine whether volume is also related to survival after hospital discharge. METHODS: Using information from the Medicare claims database, we performed a retrospective cohort study of all 7229 patients over age 65 undergoing pancreaticoduodenectomy in the United States between 1992 and 1995. We divided the study population into approximate quartiles according to their hospital's average annual volume of pancreaticoduodenectomies in Medicare patients: very low (< 1/y), low (1-2/y, medium (2-5/y), and high (5+/y). To adjust for potentially confounding variables, we used a Cox proportional hazards model to examine relationships between hospital volume and mortality, our primary outcome measure. RESULTS: Overall, 3-year survival was higher at high-volume centers (37%) than at medium- (29%), low- (26%), and very low volume hospitals (25%) (log-rank P < .0001). After excluding perioperative deaths and adjusting for case-mix, patients undergoing surgery at high-volume hospitals remained less likely to experience late mortality than patients at very low volume centers (adjusted hazard ratio 0.69, 95% CI 0.62-0.76). Relationships between hospital volume and survival after discharge were not restricted to patients with cancer diagnoses; patients with benign disease had similar improvements in late survival after surgery at high-volume centers. CONCLUSIONS: Hospital volume strongly influences both perioperative risk and long-term survival after pancreaticoduodenectomy. Our data suggest that both patient selection and differences in quality of care may underlie better outcomes at high-volume referral centers. PMID- 10455882 TI - Gender and functional outcome after coronary artery bypass. AB - BACKGROUND: Female gender is an established risk factor for increased mortality and morbidity after coronary artery bypass graft (CABG) surgery. However, the impact of gender on functional outcome after CABG is not well established. METHODS: Functional status was assessed at baseline and at 6 months with the Duke Activity Status Index (DASI) in 196 consecutive patients undergoing isolated primary CABG. Follow-up data were complete in 158 (81%) patients. The functional status of the 54 (34%) female and the 104 (66%) male patients was compared. RESULTS: The mean DASI score was significantly lower in women at baseline (19.3 +/- 13.8 vs 28.3 +/- 16.8, P = .001) and at 6 months (22.7 +/- 16.3 vs 32.8 +/- 18.2, P = .0007); however, the 6-month change in DASI score (3.3 +/- 16.9 vs 4.5 +/- 20.0, P = .7) was comparable. A similar proportion of women and men (54% vs 53%) had improved above their baseline functional level at 6 months. CONCLUSIONS: These data demonstrate that women undergo CABG at a significantly lower functional level than men; however, the functional improvement after CABG is similar across genders. PMID- 10455883 TI - Accuracy of administrative data in trauma. AB - BACKGROUND: Accurate data are needed to evaluate outcomes, therapeutics, and quality of care. This study assesses the accuracy of administrative databases in recording information about trauma patients. METHODS: Patients with thoracic aorta injury were identified with a state trauma registry, and the medical records were reviewed. Data collected were compared to administrative data on patients with thoracic aorta injuries, at the same hospitals in the same time period. RESULTS: Fifteen patients (16.3%) with thoracic aorta injury were not recorded in the administrative database, and 23 patients (18.7%) were misdiagnosed. Ninety-one patients were found in both data sources. The administrative database significantly (P < .05) underrecorded abdominal injuries (50 vs 35), orthopedic injuries (117 vs 75), and chest injuries (77 vs 48). The number of aortograms (78 vs 8), type of operative procedures (use of graft; 70 vs 30), use of bypass (35 vs 16), and complications (77 vs 33) were underreported (P < .05). The Injury Severity Score was underestimated by the administrative database (38.65 +/- 12.41 vs 25.66 +/- 9.53; P < .05). CONCLUSIONS: Administrative data lack accuracy in the recording of associated injury, injury severity, diagnostic, and procedural data. Whether these data should be used to evaluate treatment or quality of care in trauma is questionable. PMID- 10455884 TI - Neutrophils are primed for cytotoxicity and resist apoptosis in injured patients at risk for multiple organ failure. AB - BACKGROUND: Postinjury multiple organ failure (MOF) is the result of a dysregulated systemic inflammatory response in which primed neutrophils (PMNs) are sequestered in tissues, vulnerable to activation through secondary insults. Apoptosis is critical to the normal clearance of these sequestered PMNs. Conversely, dysfunctional apoptosis prolongs the PMN functional life span, potentially exacerbating PMN-mediated tissue injury and the development of MOF. We hypothesized that severe trauma, in addition to priming PMNs, provokes dysfunctional PMN apoptosis. METHODS: Neutrophils were harvested daily from 12 severely injured patients at high risk for MOF, cultured for 24 hours, and assessed for apoptosis with use of acridine orange-ethidium bromide staining and fluorescence microscopy. Priming for elastase release was measured in freshly isolated patient PMNs. Plasma from patients was assessed for its ability to delay apoptosis of normal PMNs. RESULTS: Four patients (33%) had MOF. Neutrophil apoptosis was profoundly delayed in severely injured patients throughout the 5 day study period. Priming for elastase release was augmented concomitantly. Patients' plasma delayed apoptosis of normal PMNs. CONCLUSION: In patients at high risk for postinjury MOF, PMNs are not only primed for cytotoxicity but also resist apoptosis. The dysfunctional apoptosis is attributed, at least in part, to a plasma-borne mediator. The net effect may facilitate hyperinflammatory organ injury. PMID- 10455885 TI - Thrombospondin-1 regulation of smooth muscle cell chemotaxis is extracellular signal-regulated protein kinases 1/2 dependent. AB - BACKGROUND: Thrombospondin-1 (TSP-1), an extracellular matrix protein, induces vascular smooth muscle cell (VSMC) chemotaxis. We hypothesized that extracellular signal-regulated protein kinases 1/2 (ERK1/2), a pathway of the mitogen activated protein kinase (MAPK) family, is important in TSP-1-induced VSMC chemotaxis. METHODS: A modified Boyden chamber was used to assess chemotaxis. First, a concentration curve was performed to determine the level for optimal TSP-1 induced chemotaxis. Then quiescent VSMCs were preincubated (30 minutes) in serum free medium, dimethyl sulfoxide (the inhibitor vehicle), or PD98059 (10 mumol/L, an upstream inhibitor of ERK1/2). VSMCs (50,000 cells/well) with the appropriate preincubation were placed in the top chamber. The bottom chamber contained TSP-1 (20 micrograms/mL) or serum-free medium. Results were recorded as cells/5 fields (400x). Then quiescent VSMCs were exposed to TSP-1 (20 micrograms/mL) for 0, 1, 5, 10, 30, 120, or 300 minutes. Platelet-derived growth factor (10 ng/mL) was the positive control for ERK1/2 activation. Western blot analysis was performed for activated ERK1/2. All comparisons were made by a paired t test (n = 3). RESULTS: TSP-1-induced chemotaxis peaks by a concentration of 20 micrograms/mL. PD98059 inhibited TSP-1-induced chemotaxis (P < .05). ERK1/2 was activated by TSP-1 stimulated VSMCs. CONCLUSIONS: TSP-1-stimulated VSMCs activated ERK1/2. An ERK1/2 inhibitor abolished chemotaxis, suggesting the functional importance of MAPK in TSP-1-induced VSMC chemotaxis. PMID- 10455886 TI - Integrated stimulation by CXC chemokines enhances PMN [Ca2+]i signaling in trauma and adult respiratory distress syndrome. AB - BACKGROUND: Trauma and adult respiratory distress syndrome (ARDS) are associated with increased CXC chemokine (CXC) activity. CXCs such as interleukin (IL)-8 activate polymorphonuclear neutrophils (PMNs) in the lung by means of calcium signals ([Ca2+]i). We studied CXC effects on PMN [Ca2+]i in ARDS and trauma. METHODS: Isolated PMNs were loaded with Fura-2 dye. Normal PMNs were incubated in ARDS plasma or volunteer plasma, with or without blocking antibodies to IL-8, growth-related oncogene alpha (GRO-alpha), or both (n = 6 pairs), and then stimulated with 1 to 10 nmol/L IL-8. PMNs from trauma patients or volunteers (n = 10 pairs) were stimulated with GRO-alpha, or with sequential GRO-alpha/IL-8. [Ca2+]i was measured with spectrofluorometry. RESULTS: [Ca2+]i responses to IL-8 were higher after being incubated in ARDS plasma than in volunteer plasma (251 +/ 33 vs 218 +/- 33 nmol/L, P = .03). Blockade of GRO-alpha or IL-8 reversed ARDS plasma effects. After GRO-alpha/IL-8, PMNs from trauma patients demonstrated more Ca2+ store release than did PMNs from volunteers (235 +/- 13 vs 170 +/- 10 nmol/L, P < .01). Conversely, PMNs from trauma patients lost receptor-operated Ca2+ influex to GRO-alpha. CONCLUSIONS: In traumatic ARDS, plasma CXCs prime PMNs for higher [Ca2+]i flux, making PMN activation more likely. IL-8 and GRO-alpha interact to modulate these PMN [Ca2+]i responses. PMID- 10455887 TI - Evidence of a central role for p38 map kinase induction of tumor necrosis factor alpha in pancreatitis-associated pulmonary injury. AB - BACKGROUND: Tumor necrosis factor alpha (TNF alpha) has been implicated as an important mediator in acute pancreatitis-associated adult respiratory distress syndrome, but the precise pathogenesis remains unclear. The purpose of this work was to clarify the role of TNF alpha that is produced within the lung parenchyma in the inducement of pancreatitis-related pulmonary injury and to examine 1 of the potential pathways leading to the production of pulmonary TNF alpha. METHODS: Bile salt pancreatitis was induced in rats (n = 40) that were randomized to receive a p38 mitogen-activated protein (MAP) kinase inhibitor or vehicle. A separate group (n = 16) underwent sham operation. Pulmonary capillary permeability was determined with fluorescein isothiocyanate-labeled albumin and Evans blue dye, and lung histologic analysis was performed. TNF alpha protein was measured in bronchoalveolar lavage fluid, and p38 MAP kinase was activity determined by Western blot analysis. RESULTS: The induction of pancreatitis resulted in increased pulmonary capillary leakage and worsened histologic condition (P < .01 vs sham). Effective inhibition of p38 MAP kinase-induced TNF alpha production completely prevented pancreatitis-associated pulmonary injury (P < .01 vs vehicle). CONCLUSIONS: p38 MAP kinase-induced TNF alpha production plays a central role in the development of pulmonary dysfunction, which accompanies severe acute pancreatitis in this rodent model. PMID- 10455888 TI - Inhibition of gastric cancer by camptothecin involves apoptosis and multiple cellular pathways. AB - BACKGROUND: The prognosis for gastric cancer remains dismal; novel agents that target specific molecular pathways are needed as adjuvant therapy. Camptothecin (CPT), on inhibitor of topoisomerase I, is effective in the treatment of certain solid tumors; its effects on gastric cancer are largely undefined. The purpose of this study was to (1) characterize the effects of CPT on the growth of a human gastric cancer and (2) assess potential cellular mechanisms responsible for CPT mediated growth inhibition. METHODS: The human gastric cancer SIIA was transplanted subcutaneously into athymic nude mice. After tumors reached approximately 100 mm2, mice were randomized into 3 groups to receive either CPT (5 or 10 mg/kg) or vehicle (control) intraperitoneally 3 days per week for 3 weeks; tumor size was measured biweekly. To assess potential mechanisms of CPT mediated inhibition, SIIA cells were treated with CPT (20 mumol/L) and cells were counted over a time course; apoptosis was assessed by Hoechst stain and DNA laddering. Expression of p53 (a tumor suppressor), p21Waf1 and p27Kip1 (cell cycle inhibitors), and Bcl-2 and Bcl-XL (antiapoptotic proteins) was determined. RESULTS: CPT (5 and 10 mg/kg) significantly inhibited tumor growth of SIIA gastric cancers compared with controls. CPT-mediated inhibition of SIIA cell proliferation was associated with an increase in apoptosis. Moreover, CPT treatment resulted in induction of p53, p21Waf1, and p27Kip1 and a decrease in Bcl-2 and Bcl-XL RNA and protein levels. CONCLUSIONS: Treatment with CPT effectively inhibited the growth of the human gastric cancer SIIA; the mechanism involved was induction of apoptosis mediated by up-regulation of p53, p21Waf1/Cip1, and p27Kip1 and the down-regulation of Bcl-2 and Bcl-XL. Novel agents such as CPT, which target specific molecular pathways, may prove clinically useful in the adjuvant treatment of gastric cancers. PMID- 10455889 TI - Increased myocardial calcium cycling and reduced myofilament calcium sensitivity in early endotoxemia. AB - BACKGROUND: Mechanisms of cardiac dysfunction during endotoxemia are multiple and their targets uncertain. This study tested the hypothesis that endotoxin (LPS) induces abnormal calcium-activated contractile force in the heart. METHODS: Adult rabbits were given LPS intravenously; 2 hours later hearts were studied in the Langendorff mode. Measurements included peak developed pressure (PDP), myocardial oxygen consumption (MVO2), high-energy phosphates by 31P-NMR, and beat-to-beat intracellular calcium (Cai) by fluorescence spectroscopy. Myofibrillar calcium sensitivity was assessed from the relationship of PDP to Cai and the rate of diastolic Cai removal (tau Ca) was quantified. RESULTS: Force-calcium relationships were markedly depressed in LPS hearts despite increased Cai. MVO2 was increased in parallel with increased Cai. Taken together, these data denote myofilament calcium insensitivity and mechanical inefficiency. tau Ca was markedly prolonged in LPS hearts, indicating impaired calcium reuptake and/or extrusion. High-energy phosphates and intracellular pH were unaffected by LPS; however, inorganic phosphate (Pi) was significantly increased. Dobutamine further increased Cai and MVO2 in LPS hearts without significantly improving calcium activated force. Pyruvate, an inotrope that reduces Pi, significantly improved contractility in LPS hearts. CONCLUSIONS: Endotoxemia rapidly induced futile calcium cycling and reduced myofibrillar calcium sensitivity. This state was resistant to beta-agonist inotropic stimulation; inotropes that normalize the calcium-force relationship may be more effective. PMID- 10455890 TI - Deficiency of the transcription factor c-fos increases lipopolysaccharide-induced macrophage interleukin 12 production. AB - BACKGROUND: Interleukin 12 (IL-12) p70 is a heterodimeric protein (p35, p40 subunits) that promotes T-helper TH1-type cytokine response. In critically ill patients, after severe trauma or sepsis, IL-12 production is markedly impaired. We tested the hypothesis that deficiency of the transcription factor c-fos will increase macrophage IL-12 production. METHODS: We harvested adherent peritoneal macrophages harvested from wild-type (WT), heterozygous c-fos knockout (Hetero KO), or homozygous c-fos knockout (Homo KO) mice and investigated lipopolysaccharide (LPS)-induced IL-12 p70 protein synthesis (by enzyme-linked immunosorbent assay), IL-12 p35 and IL-12 p40 messenger RNA accumulation (mRNA) (by reverse transcriptase-polymerase chain reaction), and the transcription rate (by nuclear runoff). RESULTS: (1) LPS treatment compared with vehicle increases c fos mRNA accumulation 5-fold and AP-1 DNA protein binding (electrophoretic mobility shift assay), which precedes either IL-12 p35 or IL-12 p40 mRNA accumulation. (2) LPS induces a significant increase in IL-12 p70 protein, IL-12 p40 mRNA, and the transcription rate in the Homo KO group compared with either the Hetero KO or WT groups. (3) Compared with vehicle control, we demonstrate that interferon gamma priming increases LPS-stimulated macrophage IL-12 p70 protein in the Hetero KO or WT groups to the level of the Homo KO group but has no significant effect on the Homo KO group. CONCLUSIONS: These data suggest that deficiency of the transcription factor c-fos increases LPS-induced macrophage IL 12 production, possibly by simulating the effect of interferon gamma priming. PMID- 10455891 TI - Priming of the neutrophil respiratory burst is species-dependent and involves MAP kinase activation. AB - BACKGROUND: Priming of the neutrophil respiratory burst has been implicated in the pathogenesis of multi-system organ failure (MSOF) after sepsis and trauma. The intracellular signal transduction pathways that mediate priming are unclear. METHODS: Human, porcine, rabbit, rat, and mouse neutrophils were assayed by luminol-dependent chemiluminescence in whole blood and purified neutrophil preparations. Multiple priming agents and agonists were studied, as was inhibition of priming by the p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 and the Mek 1/2 inhibitor PD98059. RESULTS: Priming by tumor necrosis factor alpha (TNF-alpha), interleukin-8 (IL-8), and granulocyte macrophage colony-stimulating factor (GM-CSF) was significantly inhibited by SB203580, whereas platelet-activating factor (PAF) priming was unaffected. Neither TNF-alpha nor PAF primed polymorphonuclear neutrophils (PMNs) within whole blood for N-formyl-methionyl-leucyl-phenylalanine (f-MLP) activation, in contrast to activation by complement-opsonized zymosan (OPZ) or low-dose phorbolmyristate acetate (PMA). Both TNF-alpha and PAF, however, primed purified neutrophils for f-MLP activation. In contrast to human and porcine PMNs, rabbit, rat, and mouse PMNs could not be primed by TNF-alpha or PAF, regardless of the final agonist. CONCLUSIONS: Priming of the PMN respiratory burst proceeds through multiple signaling pathways, depending on the particular priming agent and agonist pair. Differences in priming between PMNs in whole blood and purified preparations may be physiologically significant. There is a pronounced species dependency in the ability to prime the neutrophil respiratory burst. PMID- 10455892 TI - Nicotine's effect on hypothalamic neurotransmitters and appetite regulation. AB - BACKGROUND: Tobacco smoking reduces appetite and body weight. Cessation of smoking leads to hyperphagia and weight gain. Food intake is a function of meal number (MN) and meal size (MZ) (i.e., Food intake = MN x MZ). The effect of nicotine on these feeding components and their relationships to dopamine and serotonin in the lateral hypothalamic area (LHA) were determined. METHODS: In Fischer-344 rats the effect of 7 days of systemic nicotine infusion on the feeding patterns was measured by rat eater meter and changes in serotonin (5HT) and dopamine (DA) in the LHA were measured by in vivo microdialysis. RESULTS: Nicotine infusion caused hypophagia through a significant decrease in MN with a smaller decrease in MZ, resulting in a body weight reduction. 5HT and DA concomitantly increased in LHA. Stopping nicotine resulted in hyperphagia by a significant increase in MZ. Body weight normalized. 5HT and DA in LHA decreased after nicotine was stopped. CONCLUSION: Nicotine's hypophagic effect was associated with increased 5HT and DA in LHA, whereas hyperphagia after nicotine cessation was accompanied by decreased concentrations of the neurotransmitters. These findings suggest that nicotine affects appetite regulation, in part by modulation of LHA-DA and 5HT. PMID- 10455893 TI - Influence of oxygenation on endothelial modulation of coronary vasomotor function during hyperkalemic cardioplegia. AB - BACKGROUND: The purpose of this study was to determine the influence of oxygenation of a hyperkalemic cardioplegic solution (K-CP) on endothelial modulation of vasomotor tone and to correlate these changes with the intracellular calcium concentration ([Ca++]i) in microvascular smooth muscle. METHODS: Rat coronary arterioles were studied in a pressurized, no-flow normothermic state. Simultaneous monitoring of luminal diameter and [Ca++]i (fura 2) was performed with use of microscopic image analysis. Vessels were subjected to 60 minutes of oxygenated or hypoxic K-CP (K+ = 25.0 mmol/L) and were then reperfused with oxygenated Krebs-physiologic saline solution for 60 minutes. RESULTS: In oxygenated K-CP, the K-CP-induced contraction and [Ca++]i accumulation were significantly increased in endothelium-denuded (ED) vessels compared with endothelium-intact vessels. The effect of ED in oxygenated K-CP was mimicked by administration of the nitric oxide synthase inhibitor NG-monomethyl-L arginine. Conversely, in hypoxic K-CP the contraction was significantly attenuated in ED vessels compared with endothelium-intact vessels, although there was no significant difference in [Ca++]i. Indomethacin did not affect the endothelium-dependent contraction during hypoxic K-CP. CONCLUSIONS: Endothelium derived nitric oxide modulates the vascular tone during K-CP by regulating the vascular smooth muscle [Ca++]i, whereas endothelium-derived contracting factor(s), which is not predominantly a product of cyclo-oxygenase, may play a prominent role under hypoxic K-CP by increasing vascular smooth muscle Ca++ sensitivity. PMID- 10455894 TI - Gastrin-mediated effects of omeprazole on rat colon mucosa. AB - BACKGROUND: Omeprazole increases circulating gastrin levels, which in turn may affect the growth and differentiation of colon mucosa. Chloride transport mechanisms in normal colon were analyzed as markers for possible trophic actions of endogenous hypergastrinemia. METHODS: Four groups of Fischer rats were studied for 10 days. Group 1 (baseline) received no treatment. Group 2 received omeprazole only. Group 3 received omeprazole plus vehicle. Group 4 received omeprazole plus CCK-B gastrin receptor antagonist (GRA) L740,093 in vehicle. On day 10 serum gastrin was assayed. Colon mucosa was analyzed for protein and DNA content. Semiquantitative Northern analysis measured levels of messenger RNA (mRNA) encoding for key Cl- transporters: Na-K-Cl cotransporter (Cl- secretion in crypts), Cl-/HCO3- exchanger (Cl- absorption in villi), and Na/K adenosine triphosphatase (not directly involved in Cl- transport). RESULTS: Omeprazole increased gastrin levels, which were not altered by vehicle or GRA. Omeprazole increased protein, DNA, and Na/K adenosine triphosphatase mRNA levels, with no effect by GRA. In contrast, omeprazole decreased Na-K-Cl and Cl-/HCO3- mRNA levels, effects that were partly reversed by GRA. CONCLUSIONS: Omeprazole augments growth index values of colon mucosa independent of serum gastrin. Against a background of omeprazole-induced achlorhydria hypergastrinemia appears to influence differentiation rather than growth of normal colon mucosa. PMID- 10455895 TI - Trauma-hemorrhage delays wound healing potentially by increasing pro-inflammatory cytokines at the wound site. AB - BACKGROUND: Studies indicate impaired wound healing after trauma. The underlying mechanism remains unknown. METHODS: Mice were subjected to midline laparotomy, and polyvinyl alcohol sponges were implanted subcutaneously before hemorrhage (35 +/- 5 mmHg for 90 minutes, resuscitated) or sham operation. Wound exudate cells from the sponges were harvested on the first, third, and fifth postoperative day and cultured for 24 hours. Interleukin (IL)-1 beta, IL-6, monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-2 (MIP-2), and transforming growth factor (TGF)-beta were determined in the supernatants. IL-1 beta and IL-6 were measured in the wound fluid. RESULTS: Hemorrhage decreased collagen deposition in the wound. TGF-beta release was significantly decreased on the first and third postoperative days after hemorrhage, whereas IL-1 beta and IL-6 release was increased at 3 and 5 days after hemorrhage. Similarly, IL-1 beta and IL-6 in the wound fluid were significantly increased at 3 days after hemorrhage. CONCLUSIONS: Because increased levels of pro-inflammatory cytokines and decreased amounts of TGF-beta have been reported to impair the process of wound healing, the increased release of IL-1 beta and IL-6 and the decreased release of TGF-beta after hemorrhage might contribute to the decreased collagen production in those animals. Thus, attempts to locally change the ratio of those cytokines in trauma victims might be useful for improving wound healing in those patients. PMID- 10455896 TI - Delayed treatment with diethyl maleate prevents E-selectin expression in human endothelial cells. AB - BACKGROUND: Polymorphonuclear leukocyte (PMN) infiltration is a significant contributor to tissue damage in many disease states and is known to occur through an orderly set of events. The endothelial cell adhesion molecule E-selectin is involved in the initial rolling of PMNs on the endothelium at sites of inflammation. We have previously shown that the glutathione depleting agent diethyl maleate (DEM) attenuates lung injury in a rodent model of intratracheal LPS stimulation. We hypothesized that DEM might attenuate E-selectin in LPS treated human umbilical vein endothelial cells as a mechanism underlying this effect. Further, we investigated the role of delayed treatment with DEM on E selectin expression. METHODS: Human umbilical vein endothelial cells were treated with DEM (100 to 400 mumol/L) before or after LPS stimulation (1 microgram/mL). Surface expression of E-selectin was examined using a cellular enzyme-linked immunosorbent assay. E-selectin mRNA transcripts were detected by Northern blot analysis. Nuclear factor-kappa B (NF-kappa B) activity was detected with gel shift assays. RESULTS: DEM significantly inhibited LPS-induced E-selectin surface expression and mRNA levels in a dose-dependent fashion, with complete inhibition at 250 mumol/L, without affecting cell viability. This inhibitory effect was seen even if DEM was added up to 60 minutes after LPS. DEM inhibited NF-kappa B nuclear translocation in a manner that mirrored protein and mRNA levels. CONCLUSIONS: Delayed treatment with DEM attenuates NF-kappa B nuclear translocation and E-selectin expression in human umbilical vein endothelial cells up to 60 minutes after the onset of LPS stimulation. Thus, DEM may represent an effective intervention for PMN-mediated organ injury even when given after an inflammatory insult. PMID- 10455897 TI - Short-chain fatty acids and thyroid hormone interact in regulating enterocyte gene transcription. AB - BACKGROUND: Enterocyte differentiation is known to be regulated by a variety of extracellular compounds, among which are triiodothyronine (T3) and the short chain fatty acids (SCFAs). Because several SCFAs are known to induce histone hyperacetylation, and T3 action has been recently linked to chromatin structure, we sought to investigate the interplay between SCFAs and T3 in regard to the enterocyte differentiation marker, intestinal alkaline phosphatase (IAP). METHODS: Caco-2 cells were transiently transfected with a reporter construct containing 2.4 kb of the human IAP gene 5' flanking region (IAP2.4CAT). Cotransfections were carried out with and without thyroid hormone receptor-1 (TR beta-1) or histone deacetylase-1 (HDAC-1) expression plasmids. Cells were treated with 5 mmol/L SCFAs (propionic, butyric, valeric, or caproic acids as propionate, butyrate, valerate, or caproate, respectively), with and without 10 nmol/L T3. Reporter gene activity was measured and the level of histone acetylation assessed by means of acid-urea-triton (AUT) gel assays. RESULTS: TR beta-1 cotransfection caused a marked decrease in IAP reporter gene activity, which is consistent with the well-known phenomenon of ligand independent repression (LIR), whereas T3 treatment reversed the LIR and caused further reporter gene activation. Treatment with SCFAs similarly resulted in a complete blockage of LIR, and, in fact, turned the TR beta-1 into a transcriptional activator, even in the absence of T3. Concomitant treatment with T3 and butyric acid produced an additive effect on IAP transactivation. In contrast, cotransfection with HDAC-1 attenuated the effects of SCFAs on IAP gene activation. AUT gel studies demonstrated histone hyperacetylation in response to SCFA treatment. CONCLUSION: One or more DNA cis elements in the human IAP gene mediate ligand independent repression by the TR beta-1, an effect that can be entirely reversed by those SCFAs that induce histone hyperacetylation. In addition T3 and SCFAs can act in concert to induce IAP gene transcription, demonstrating an important link between triiodothyronine and histone hyperacetylation in regard to enterocyte-specific gene expression. PMID- 10455898 TI - Anoikis, extracellular matrix, and apoptosis factors in isolated cell transplantation. AB - BACKGROUND: Isolated cell transplantation (ICT) of pancreatic islets (PI), nerve tissue, hepatocytes, and other cells is an exciting new concept of transplantation. PI transplantation can be successful in reversing diabetes but, unlike whole pancreas, grafts have a unique and unexplained high failure rate, with over 60% loss by 3 to 6 months. We established that PI of rhesus monkeys have a high rate of death within 48 hours of isolation as a result of apoptosis, as measured with the Annexin V assay (Pharmingen, San Diego, Calif). In contrast, PI incompletely separated from the extracellular matrix (ECM) remained viable for prolonged periods in culture and performed superiorly in perifusion assays (insulin secretion of 4.6 +/- 0.8 times basal secretion). METHODS: We studied the ability of the anti-AP Bcl-2 molecule, known to block anoikis (a mechanism of AP due to cell-ECM separation), to prevent apoptosis of isolated PI. RESULTS: PI transduced with an adenovirus-Bcl-2 gene complex showed a high viability and a low AP rate in culture versus control rhesus PI. CONCLUSION: In summary, PI protected from AP by a surrounding ECM mantle or by Adenovirol Vector (AdV) transduction of the Bcl-2 gene showed superior viability without AP in vitro and in vivo evidence of a preserved insulin secretion response to glucose. PMID- 10455899 TI - Granulocyte colony-stimulating factor and neutrophil-related changes in local host defense during recovery from shock and intra-abdominal sepsis. AB - BACKGROUND: We have reported that treatment with exogenous granulocyte colony stimulating factor (G-CSF) improves abscess localization and reduces mortality without aggravating neutrophil (PMN)-mediated reperfusion injury in a model of septic abdominal trauma. The purpose of this study was to determine actions of G CSF on PMN function in the peritoneum. METHODS: Anesthetized swine were pretreated with broad-spectrum antibiotics and underwent cecal ligation and incision and 35% hemorrhage (trauma). After 1 hour they were resuscitated with shed blood, crystalloid, and either G-CSF (n = 10) or saline solution vehicle (n = 9). The animals were observed for 72 hours. RESULTS: After trauma, saline solution treatment increased PMN infiltration into the peritoneum within 2 hours (P = .035), increased peritoneal PMN elastase production (i.e., cytotoxicity) by 24 hours (P = .004), and decreased adherence of peritoneal PMNs to an artificial substrate from 4 to 72 hrs (P = .043). The mean autopsy score was 7.0 +/- 0.5. With G-CSF treatment peritoneal neutrophilia was enhanced (maximum 48 hours, P = .002) and PMN cytotoxicity was augmented and delayed (maximum 48 hours, P = .004). Despite these changes, adherence of peritoneal PMNs was not significantly changed and there was no evidence for PMN-mediated damage in the lung as judged by bronchoalveolar lavage protein, bronchoalveolar lavage PMNs, lung tissue myeloperoxidase, or histologic changes. The mean autopsy score was improved to 4.1 +/- 0.3 (P < .001). CONCLUSIONS: G-CSF in resuscitation fluids improved localization of an intra-abdominal septic focus by increased production of circulating PMNs, increased PMN extravasation into the peritoneal cavity, and increased PMN cytotoxicity at the abdominal septic focus, without exaggerating PMN-dependent reperfusion injury in the lung. Therefore these data further support the idea that G-CSF in resuscitation fluids might reduce septic complications in the multiply injured trauma patient. PMID- 10455900 TI - Dopamine induces neutrophil apoptosis through a dopamine D-1 receptor-independent mechanism. AB - BACKGROUND: For the normal resolution of an acute inflammatory response, neutrophil (PMN) apoptosis is essential to maintain immune homeostasis and to limit inappropriate host tissue damage. A delay in PMN apoptosis has been implicated in the pathogenesis of the systemic inflammatory response syndrome (SIRS). Dopamine, a biogenic amine with known cardiovascular and neurotransmitter properties, is used in patients with SIRS to maintain hemodynamic stability. We sought to determine whether dopamine may also have immunoregulatory properties capable of influencing PMN apoptosis, function, and activation state in patients with SIRS. METHODS: PMNs were isolated from healthy volunteers and patients with SIRS and treated with varying doses of dopamine and a dopamine D-1 receptor agonist, fenoldopam. PMN apoptosis was assessed every 6 hours with use of propidium iodide DNA staining and PMN function was assessed with use of respiratory burst activity, phagocytosis ability, and CD11a, CD11b, and CD18 receptor expression as functional markers. RESULTS: There was a significant delay in PMN apotosis in patients with SIRS compared with controls. Treatment of isolated PMNs from both healthy controls and patients with SIRS with 10 and 100 mumol/L dopamine induced apoptosis. PMN ingestive and cytocidal capacity were both decreased in patients with SIRS compared with controls. Treatment with dopamine significantly increased phagocytic function. Fenoldopam did not induce PMN apoptosis. CONCLUSION: Our data demonstrate for the first time that dopamine induces PMN apoptosis and modulates PMN function both in healthy controls and in patients with SIRS. These results indicate that dopamine may be beneficial during SIRS through a nonhemodynamic PMN-dependent proapoptotic mechanism. PMID- 10455901 TI - Optimization of ex vivo inducible nitric oxide synthase gene transfer to vein grafts. AB - BACKGROUND: Vein graft failure as the result of intimal hyperplasia (IH) remains a significant clinical problem. Ex vivo modification of vein grafts using gene therapy is an attractive approach to attenuate IH. Gene transfer of the inducible nitric oxide synthase (iNOS) gene effectively reduces IH. However, iNOS activity after gene transfer may be impaired by the availability of cofactor, such as tetrahydrobiopterin (BH4). The purpose of this study is to determine the optimal conditions for ex vivo adenoviral-mediated iNOS gene transfer into arterial and venous vessels. METHODS: Porcine internal jugular veins and carotid arteries were infected ex vivo with the adenoviral iNOS vector (AdiNOS) and with an adenovirus carrying the cDNA encoding guanosine triphosphate cyclohydrolase I (AdGTPCH), the rate-limiting enzyme for BH4 synthesis. The production of nitrite, cyclic guanosine monophosphate (cGMP), and biopterin were assessed daily. RESULTS: Nitric oxide (NO) production after iNOS gene transfer was maximal when vessels were cotransduced with AdGTPCH. NO production in these vessels persisted for more than 10 days. Vein segments generated approximately 2-fold more nitrite, cGMP, and biopterin than arterial segments infected with AdiNOS/AdGTPCH. Submerging vein segments into adenoviral solution resulted in improved gene transfer with greater nitrite and cGMP release compared with infections carried out under pressure intraluminally. Similarly, injury to the vein segments before infection with AdiNOS resulted in less nitrite production. CONCLUSIONS: These data demonstrate that AdiNOS can efficiently transduce vein segments ex vivo and that the cotransfer of GTPCH can optimize iNOS enzymatic activity. This cotransfer technique may be used to engineer vein grafts before coronary artery bypass to prevent IH. PMID- 10455902 TI - Acquisition of surgical skills: a randomized trial of didactic, videotape, and computer-based training. AB - BACKGROUND: Although computer-based training (CBT) can enhance didactic instruction, few studies have assessed the efficacy of CBT for basic surgical skills training. This study compares CBT with traditional methods of basic surgical skills training. METHODS: Sixty-nine naive medical students were randomized into 3 treatment groups for basic surgical skills instruction: didactic, videotape, or CBT. All instructional material contained the same pictures, text, and audio. With use of a multiple-choice question examination and a series of performance stations, students were objectively assessed before, immediately after, and 1 month after skills instruction. Raters were blinded to treatment modality during the follow-up evaluation. RESULTS: There were no significant differences among the groups before treatment. After treatment, the didactic group scored higher on the multiple-choice question examination. In contrast, the videotape and CBT groups demonstrated statistically significant (P < .01) enhancement of technical skills compared with the didactic group. After 1 month, a calculated performance quotient revealed statistically significant (P < .01) improvement only in the CBT group. The amount of time students spent practicing their skills was not significantly different among the groups. CONCLUSIONS: CBT is as effective as, and possibly more efficient, than traditional methods of basic surgical skills training for medical students. PMID- 10455903 TI - Intern call schedules and their relationship to sleep, operating room participation, stress, and satisfaction. AB - BACKGROUND: The ideal resident call schedule remains unknown. This study assessed the impact of different call schedules on intern performance and education. METHODS: A year-long, prospective, observational study of first-year residents in a surgery training program was performed with use of intern sleep/operative logs and questionnaires, and faculty questionnaires. RESULTS: Compared with interns taking call every third or fourth night (and cross-covering a separate service), interns taking call every other night reported the greatest amount of fatigue and stress, the lowest satisfaction, and the fewest operative cases. Errors in patient care were not different between schedules. Multivariate analysis revealed that operative participation was inversely related to frequency of night call and level of fatigue post call, stress was related to fatigue while off call and service census, and overall satisfaction was associated with infrequency of call and operative cases performed. Faculty reported more errors by interns cross covering other services and less operating room participation by interns taking call every other night. CONCLUSIONS: No single resident schedule optimally balances patient care and resident education and satisfaction. All 3 patterns of call studied are acceptable; specific decisions regarding the allocation of house staff manpower should be flexible and dependent on individual service and educational needs. PMID- 10455904 TI - Intestinal ischemia-reperfusion injury is mediated by the membrane attack complex. AB - BACKGROUND: The dependence of intestinal ischemia-reperfusion injury on the classical complement pathway has been shown with the complement antagonist (sCR1) and complement-specific knockout mice. Using C5 deficient mice, we show that the membrane attack complex mediates local injury. METHODS: Mice underwent intestinal ischemia-reperfusion. Albumin leak and histologic evidence were compared in wildtype mice, wildtypes treated with sCR1, neutrophil-depleted wildtypes, C5 deficient mice, and C5-deficient mice reconstituted with wildtype serum. Neutrophil tissue levels in injured C5-deficient and wildtype intestines were compared. RESULTS: C5-deficient mice had a reduction in injury similar to mice treated with sCR1. Injury was restored by reconstitution with wildtype serum. Wildtype injury was unaffected by neutrophil depletion. Injured intestines of C5 deficient and wildtype mice had similar neutrophil levels. Immunohistochemistry of wildtype and reconstituted C5-deficient mice demonstrated injured intestinal epithelium although C5-deficient mice and sCr1-treated mice were similar to sham mice. CONCLUSIONS: C5-deficient animals are protected from local injury. Injury is unaffected by neutrophil depletion, and the presence of neutrophils in injured tissue is independent of C5. Local injury is C5 dependent, but the action of C5a on granulocytes is not required. Therefore the membrane attack complex mediates local injury. PMID- 10455905 TI - Discordant tumor necrosis factor-alpha superfamily gene expression in bacterial peritonitis and endotoxemic shock. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is a member of a large family of predominantly homotrimeric type II membrane-associated proteins with both proinflammatory and apoptosis-inducing properties. Although TNF-alpha expression has been studied extensively, little is known about the expression of other members of the TNF-alpha superfamily during acute inflammatory processes. METHODS: TNF-alpha, Fas ligand (FasL), and TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) messenger RNA (mRNA) expression were examined in liver, lung, spleen, and kidney after either a cecal ligation and puncture or endotoxemic shock with use of semiquantitative reverse transcriptase-polymerase chain reaction. RESULTS: Cecal ligation and puncture increased TNF-alpha mRNA in lung and liver (both P < .05) within 3 hours, which was paralleled by increased FasL mRNA. In the spleen TNF-alpha and FasL mRNA significantly declined (both P < .05). In contrast to TNF-alpha and FasL, TRAIL mRNA levels were unchanged in all organs except lung, where it was reduced at 24 hours (P < .05). Endotoxemic shock also increased lung TNF-alpha and FasL mRNA levels (both P < .05). CONCLUSIONS: In acute inflammatory processes TNF-alpha and FasL mRNA increase concordantly in several solid organs. In contrast, TRAIL mRNA levels do not consistently change during these acute inflammatory processes, suggesting that its expression is under independent and discordant regulatory control. PMID- 10455906 TI - Importance of timing and length of administration of angiogenesis inhibitor TNP 470 in the treatment of K12/TRb colorectal hepatic metastases in BD-IX rats. AB - BACKGROUND: The timing and length of administration of angiogenesis inhibitor TNP 470 was altered to evaluate the effect on disease progression in a rat model of colorectal hepatic metastases. METHODS: Pair-fed BD-IX rats, injected intrasplenically with rat colon adenocarcinoma K12/TRb cells at day 0, were randomized to receive subcutaneous injections of either placebo or 15 mg/kg TNP 470 on alternate days: for 2 weeks beginning 24 hours after tumor inoculation ("Early"), for 4 weeks beginning 24 hours after tumor inoculation ("Prolonged"), or for 2 weeks beginning at day 15 after macroscopic tumor nodules were confirmed ("Delayed"). Response to treatment was evaluated by counting tumor nodules on the surface of the liver at laparotomy on day 14 and 28 after tumor inoculation. The animals were followed for survival and cause of death. RESULTS: Maximal suppression of hepatic metastases at day 28 required 4-week rather than 2-week TNP-470 administration. Prolonged TNP-470 administration resulted in significantly fewer hepatic metastases at day 28 compared to control (P < .05). Early and prolonged TNP-470 improved survival (Wilcoxon test, P < .05) compared with delayed TNP-470 and placebo. Delayed TNP-470 administration did not increase survival or significantly diminish the number of metastases at day 28 compared with placebo. CONCLUSIONS: These data suggest that prolonged adjuvant antiangiogenic therapy may suppress colorectal hepatic micrometastases. PMID- 10455907 TI - Cyclooxygenase-2 alters transforming growth factor-beta 1 response during intestinal tumorigenesis. AB - BACKGROUND: Recent investigation suggests that cyclooxygenase-2 plays an important role in colorectal carcinogenesis. Transforming growth factor-beta1 (TGF-beta 1) is one of the most potent stimulators of cyclooxygenase-2 expression. A key step in intestinal tumorigenesis involves alteration of the normal cellular response to TGF-beta 1. We have hypothesized that overexpression of cyclooxygenase-2 alters intestinal epithelial response to TGF-beta 1. METHODS: RIE-1 cells were stably transfected with rat cyclooxygenase-2 complementary DNA in either the sense (RIE-S) or antisense (RIE-AS) orientation. Tumor cell invasion was assessed with a modified Boyden collagen type I invasion assay in the presence of TGF-beta 1, antibody to urokinase plasminogen activator (uPA), or the selective cyclooxygenase-2 inhibitor SC-58125. Expression of uPA, uPA receptor, and plasminogen activator inhibitor-1 were determined by Western blot and enzyme-linked immunosorbent assay. RESULTS: RIE-1 and RIE-AS did not invade although RIE-S cells were minimally invasive at baseline. TGF-beta 1 had no effect on RIE-1 or RIE-AS invasion; however, TGF-beta 1 significantly upregulated RIE-S cell invasion. All 3 RIE cell lines produce minimal uPA under basal conditions. TGF-beta 1 upregulated uPA production only in the RIE-S cells. Both antibody to uPA and SC-58125 reversed TGF-beta-mediated RIE-S cell invasion. SC 58125 inhibited TGF-beta-mediated RIE-S uPA production. CONCLUSIONS: These results demonstrate that overexpression of cyclooxygenase-2 alters intestinal epithelial response to TGF-beta 1, which may be a mechanism by which cyclooxygenase-2 promotes colon carcinogenesis. PMID- 10455908 TI - Ubiquitin-proteasome inhibition enhances apoptosis of human pancreatic cancer cells. AB - BACKGROUND: Tumor necrosis factor (TNF-a)-induced apoptosis is limited by coactivation of nuclear factor kappa B (NF-kb)-dependent antiapoptotic genes. Nuclear translocation of NF-kB requires degradation of ubiquitinated phospho-IkB a by the 26S proteasome. We examined whether inhibition of the ubiquitin proteasome pathway enhances TNF-a-induced apoptosis in BxPC-3 human pancreatic cancer cells. METHODS: Serum-starved BxPC-3 cells (12 hours) were pretreated or not for 50 minutes with PSI (30 m mol/L), a peptide aldehyde known to inhibit specifically the chymotrypsin-like activity of the 26S proteasome. Cells were subsequently stimulated with recombinant human TNF-a (400 units/mL). Western blots were performed using antibodies to IkB-a and phospho-IkB-a. Level of apoptosis was determined by two methods: enzyme-linked immunosorbent assay detection of interhistone DNA fragments and flow cytometry with propidium iodide staining. RESULTS: TNF-a-induced degradation of IkB-a was inhibited by PSI. Phospho-IkB-a accumulation was observed 20 minutes after TNF-a stimulation. Apoptosis relative to constitutive levels was significantly increased after PSI pretreatment, as measured by DNA fragmentation (P < or = .05 by Student t test). Percent apoptosis by flow cytometry confirmed marked increases in apoptotic cell fractions from 5.9% (untreated) to 6.8% (TNF-a alone), 16.4% (PSI alone), and 18.9% (PSI and TNF-a). CONCLUSIONS: PSI enhances both constitutive and TNF-a induced apoptosis through inhibition of IkB-a degradation in BxPC-3 human pancreatic cancer cells. PMID- 10455909 TI - What is the role of interleukin 10 in polymicrobial sepsis: anti-inflammatory agent or immunosuppressant? AB - BACKGROUND: Controversy exists concerning the role of interleukin 10 (IL-10) in sepsis. When IL-10 is used in models of endotoxemia, it appears to protect (by anti-inflammatory effects), whereas in models of polymicrobial sepsis it seems to be deleterious (by immunosuppression?). However, little direct evidence for such an immunosuppressive role is available for polymicrobial sepsis. Thus the aim of this study was to determine whether IL-10 contributes to lymphocyte immunosuppression in a model of cecal ligation and puncture (CLP) and whether neutralization of IL-10 has any salutary effects on survival after sepsis. METHODS: To assess the former, polymicrobial sepsis was induced in male C57BL/6J wild-type (+/+) and C57BL/6J-IL-10 knockout(-/-) mice by CLP. Splenocytes were harvested 24 hours later and stimulated with concanavalin A to assess their proliferative capacity and their ability to release the Th1 lymphokines interleukin 2 and interferon gamma (by enzyme-linked immunosorbent assay, nanograms/millilter). To further verify the immunosuppressive role of IL-10, splenocytes were obtained from male C3H/HeN mice 24 hours after CLP and then stimulated in the presence or absence of anti-IL-10 monoclonal antibody (Mab, 4 micrograms/mL). To assess the in vivo effects of IL-10 neutralization on survival after CLP, C3H/HeN mice (16 per group) were given 250 micrograms of anti-IL-10 Mab (intraperitoneally) either immediately after CLP (before the initiation of the hyperdynamic phase) or 12 hours after CLP (the beginning of the hypodynamic state). Control mice were given nonspecific rat immunoglobulin G. RESULTS: These data indicate that IL-10 deficiency (-/-) prevents the depression of the proliferative capacity and Th1 lymphokine production after sepsis. Analysis of the interleukin 2-interferon gamma production patterns and proliferative capacity in lymphocytes treated with anti-IL-10 Mab confirmed the role of IL-10 in suppressing lymphocyte responsiveness in CLP. Interestingly, however, only delayed administration (12 hours after CLP) of anti-IL-10 markedly increased survival of mice (Fisher's exact test, P < .05). CONCLUSION: The results not only illustrate IL-10's role in septic immune dysfunction but document that anti-IL-10 administration beyond the initial proinflammatory hyperdynamic state of polymicrobial sepsis improves survival of animals subjected to sepsis. PMID- 10455910 TI - Long-term survival of an extremity composite tissue allograft with FK506 mycophenolate mofetil therapy. AB - BACKGROUND: High-dose tacrolimus (FK506) monotherapy has significantly prolonged rat hindlimb allograft survival. With an eye toward direct clinical application, we used a large-animal extremity composite tissue allograft model to assess the antirejection efficacy and systemic toxicity of combination FK506-mycophenolate mofetil (MMF) treatment. METHODS: Radial forelimb osteomyocutaneous flap transplants were performed between size-matched outbred pigs assigned to one of two groups: 5 control pigs received no immunosuppression and 9 animals received a once-daily oral FK506-MMF-prednisone regimen. Rejection was assessed by visual inspection of flap skin and was correlated with serial histopathologic examination of skin biopsy specimens. RESULTS: In all control pigs the flap was completely rejected on day 7. Of the 9 pigs receiving treatment, 3 died from pneumonia on days 29, 30, and 83 without signs of rejection and another died from gastric rupture on day 42 with persistent mild rejection. The remaining 5 animals were free of rejection at the end of the 90-day follow-up period (P < 0.005 vs controls). Overall, 5 pigs had pneumonia, 4 septic arthritis, 3 toe abscesses, and 5 diarrhea and decreased weight gain. CONCLUSIONS: Combination oral FK506-MMF treatment provided a superior antirejection effect but more produced more toxicity than that previously demonstrated with cyclosporin A-MMF therapy in our model. Our results suggest that reduction of FK506 or MMF doses might decrease both infectious and drug-specific side effects while still providing adequate prophylaxis against rejection. PMID- 10455911 TI - Proinflammatory cytokines (tumor necrosis factor and interleukin 1) stimulate release of high mobility group protein-1 by pituicytes. AB - BACKGROUND: Cytokines mediate the metabolic and physiologic responses to injury and infection. Anterior pituitary cells express receptors for tumor necrosis factor (TNF) and interleukin 1 (IL-1), which can signal these cells to release corticotropin, growth hormone, and cytokines such as IL-1 and macrophage migration inhibitory factor. This interaction provides an important link between the immune system and the neuroendocrine system. We reasoned that pituicytes activated with TNF or IL-1 might release previously unrecognized factors that could participate in this signaling from the neuroendocrine to the immune system. METHODS: Proteins released from rat pituicytes (GH3) after stimulation with proinflammatory cytokines were identified by N-terminal amino acid sequencing. Polyclonal antibodies against a peptide corresponding to the N-terminal amino acid sequence were generated and used to determine the kinetics of protein release. RESULTS: Cytokine stimulation induced the release of a 30-kd protein from rat pituicytes. After the protein was isolated and the N-terminal amino acid sequence determined, a protein database analysis revealed that it is high mobility group-1 (HMG-1) protein. TNF and IL-1 induced the release of HMG-1 from pituicytes in a time- and dose-dependent manner. Interferon gamma alone did not induce the release of HMG-1, but it enhanced TNF-induced HMG-1 release. CONCLUSION: Stimulation of pituicytes by TNF or IL-1 induces the release of HMG 1, which may participate in the regulation of neuroendocrine and immune responses to infection or injury. PMID- 10455912 TI - Structural and functional changes resulting from islet isolation lead to islet cell death. AB - BACKGROUND: Islet isolation exposes the islet to a variety of cellular stresses and disrupts the cell-matrix relationship--events known to be associated with apoptosis. The purpose of this study was to determine whether islet isolation leads invariably to islet cell death and to specify the mechanisms involved. METHODS: Canine islets were isolated using Liberase CI and purified using a centrifuge. Islets were sampled for up to 5 days in culture and analyzed by routine histology, electron microscopy, immunocytochemistry, and reticulin staining for basement membrane. Apoptosis was assessed by cell death enzyme linked immunosorbent assay and terminal deoxynucleotidyl transferase-mediated decoxyuridine triphosphate nick and labeling (TUNEL) assay. Activation of the prosurvival ERK1/2 and proapoptotic p38 and JNK were determined by immunoblotting. RESULTS: Immediately after isolation, the peri-insular basement membrane was absent, and integrin-alpha 5 expression diminished. DNA fragmentation rose from 2.5 +/- 1.8 (arbitrary units) on the day of isolation to 42.4 +/- 6.7 48 hours later (P < .05), coinciding with the appearance of pyknotic nuclei and apoptotic bodies. The apoptotic index determined by TUNEL assay increased from 5% +/- 1% on the day of isolation to 60% +/- 2% on day 5 (P < .01), and most of the affected cells were beta-cells. Finally, the p38 and JNK activity were elevated relative to ERK1/2. CONCLUSIONS: During isolation, islet cells undergo profound changes in structure and function, resulting in beta-cell apoptosis. These findings suggest that strategies directed to the manipulation of the cell-matrix relationship and the modulation of mitogen-activated protein kinase signal transduction may offer a valuable new approach to improving islet transplant outcome. PMID- 10455913 TI - Inhibition of the transcription factor nuclear factor-kappa B by adenoviral mediated expression of I kappa B alpha M results in tumor cell death. AB - BACKGROUND: The activation of transcription factor nuclear factor-kappa B (NF kappa B) by extracellular stimuli has been shown to protect cells from apoptotic cell death. Inhibition of NF-kappa B activity should result in increased tumor cell killing in response to apoptotic stimuli. This study evaluated the effect of inhibition of NF-kappa B on a series of sarcoma and normal cell lines. METHODS: Human sarcoma cell lines (HT1080, SKLMS-1, and MFH) and normal cell lines (NLF and BSMC) were infected with an adenoviral dominant-negative mutant Ad5I kappa B alpha M in vitro. Control cells were infected with the empty adenoviral vector and mock-infected with media alone. Viable cell counts were determined by microscopic evaluation on days 1 to 6 after infection. Cell proliferation was determined at 48 hours by MTT (1-[4,5-dimethylthiazol-2-yl]-3,5-dephenylformazan) assay. RESULTS: All cell lines showed evidence of successful adenoviral infection as evidenced by the infection of all cell lines with the adenoviral marker gene Ad5 LacZ. All the tumor cells were found to have a significant decrease in cell viability and proliferation after treatment with the Ad5I kappa B alpha M gene compared with both mock-infected cells and cells infected with empty vector (P < .0001). The normal cell lines, although able to be successfully infected, did not show a significant decrease in cell viability or proliferation with adenoviral mediated I kappa B alpha M infection. CONCLUSIONS: Inhibition of NF-kappa B through adenoviral-mediated infection of the dominant-negative inhibitor I kappa B alpha M resulted in a significant decrease in tumor cell viability and proliferation while having no deleterious effect on normal cell lines. The Ad5I kappa B alpha M gene therefore could be potentially used as a clinical treatment for patients with soft-tissue sarcoma. PMID- 10455914 TI - Mitogen-activated protein kinases signal inhibition of apoptosis in lipopolysaccharide-stimulated neutrophils. AB - BACKGROUND: Neutrophil (PMN) apoptosis is critical to the resolution of infection and the limitation of inflammation. Bacterial endotoxin (lipopolysaccharide [LPS]) inhibits PMN apoptosis and activates the p38 mitogen-activated protein kinase (MAPK) signal cascade. The role of p38 and other MAPKs (ERK and SAPK/JNK) in regulating PMN apoptosis after LPS stimulation is unknown. We hypothesize that MAPK activation by LPS signals inhibition of PMN apoptosis. METHODS: PMNs were isolated from the blood of healthy human volunteers and incubated with PD98059 (ERK inhibitor), SB203580 (p38 inhibitor), or 0.1% dimethyl sulfoxide (vehicle) for 1 hour before treatment with LPS (0, 10, or 1000 ng/mL). Neutrophil MAPK activation was determined by Western blot analysis for phosphorylated p38, ERK, and SAPK/JNK. Apoptosis was quantified by flow cytometry with use of propidium iodide and annexin V. RESULTS: LPS inhibited PMN apoptosis and activated p38 and ERK in a dose- and time-dependent fashion. SAPK/JNK was not activated by LPS. Treatment of cells with ERK inhibitor before LPS stimulation abrogated LPS signaled inhibition of PMN apoptosis. Conversely, p38 inhibition with SB203580 augmented inhibition of apoptosis by LPS. CONCLUSIONS: These data demonstrate opposing roles of MAPKs in mediating PMN apoptosis after LPS stimulation. We conclude that LPS signal transduction by ERK inhibits PMN apoptosis while activation of p38 promotes apoptosis. PMID- 10455915 TI - Synergistic antitumor effects of HER2/neu antisense oligodeoxynucleotides and conventional chemotherapeutic agents. AB - BACKGROUND: The HER2/neu oncogene is overexpressed in a substantial fraction of human tumors. HER2/neu overexpressing tumors may be intrinsically resistant to chemotherapy. The present study examined the ability of antisense-mediated downregulation of HER2/neu expression to enhance the antitumor effects of conventional chemotherapeutic agents against human tumor cells that overexpress HER2/neu. METHODS: The effects of HER2/neu antisense oligodeoxynucleotides (ODNs) on the growth inhibitory and proapoptotic activity of several distinct chemotherapeutic agents were examined in vitro. In vivo effects of HER2/neu antisense ODNs in combination with doxorubicin hydrochloride were assessed by examining the growth of human tumor xenografts implanted into nude mice. RESULTS: The proliferation of tumor cell lines that overexpress HER2/neu was inhibited by antisense ODNs in combination with conventional chemotherapeutic agents in an additive or synergistic fashion. Such combination therapy also demonstrated synergistic activation of apoptosis. HER2/neu antisense ODNs in combination with doxorubicin hydrochloride demonstrated synergistic antitumor effects in vivo as well. CONCLUSIONS: Downregulation of HER2/neu expression can enhance the sensitivity of human cancer cells, which overexpress HER2/neu to the cytotoxic effects of chemotherapy. Antisense ODNs targeting the HER2/neu gene may play a role in cancer therapy. PMID- 10455916 TI - Mechanical endothelial damage results in basic fibroblast growth factor-mediated activation of extracellular signal-regulated kinases. AB - BACKGROUND: Endothelial damage, such as that associated with balloon angioplasty or preparation of veins for bypass grafts, results in intimal hyperplasia. Growth factors and cytokines that modulate endothelial cell functions through various intracellular signaling pathways mediate rapid endothelial repair, which may prevent or reduce restenosis. Here we investigated the effect of mechanical injury of endothelial cells on the mitogen-activated kinase signaling pathways, extracellular-signal-regulated kinases (ERKs), C-Jun N-terminal kinase (JNK/SAPK), and p38. METHODS: Confluent human umbilical vein endothelial cells or bovine aortic endothelial cells were wounded with a razor blade; mitogen activated kinase activation was monitored by immunoblotting with antibodies to active ERK, JNK/SAPK, or p38. RESULTS: Wounding of human umbilical vein endothelial cell or bovine aortic endothelial cell monolayers resulted in rapid (5-minute) activation of ERK-1 and -2, which was abolished by monoclonal antibody to basic fibroblast growth factor (FGF-2). This antibody or an inhibitor of ERK activation, PD98059, also blocked endothelial cell migration after the wounding. Thus FGF-2-induced ERK activation mediates the endothelial response to wounding. CONCLUSIONS: ERK-1 and -2 are activated by FGF-2 released from endothelial cells in response to injury. Therapeutic strategies aimed at reducing FGF-2-induced intimal hyperplasia should preserve ERK activation in endothelial cells while abolishing it in smooth muscle cells. PMID- 10455917 TI - Macrophage depletion alters vein graft intimal hyperplasia. AB - BACKGROUND: The principal cause of vein graft failure is intimal hyperplasia (IH); however, its etiology remains unclear. In a rat model of vein graft IH we have observed prolonged transmural macrophage infiltration, leading us to hypothesize that these cells regulate IH. To test this, we used liposome encapsulated dichloromethylene bisphosphonate (L-Cl2MBP) to deplete rat macrophages and observed the effects on IH. METHODS: Epigastric vein-to-femoral artery grafts were microsurgically placed in male Lewis rats that had been intravenously injected with L-Cl2MBP, phosphate-buffered saline solution liposomes, or phosphate-buffered saline solution alone 2 days before surgery. Several animals in each group received a second equivalent dose at 2 weeks. Grafts, contralateral epigastric veins, spleens, and livers were harvested at 1, 2, and 4 weeks for histologic examination, immunohistochemistry, and transmission electron microscopy. RESULTS: In the L-Cl2MBP-treated animals splenic and hepatic macrophages were greatly reduced, confirming the efficacy of the agent. At 1 to 2 weeks graft macrophages were significantly decreased, and there was a trend toward decreased IH. At 4 weeks macrophage numbers were normal and IH development had resumed. In contrast, the 4-week grafts treated with 2 doses of L-Cl2MBP had fewer macrophages and displayed severely attenuated IH. CONCLUSIONS: The results indicate a suppression of IH as macrophages are depleted, with a resumption of the process as macrophages repopulate the graft. PMID- 10455918 TI - Inducible nitric oxide synthase (iNOS) gene deficiency increases the mortality of sepsis in mice. AB - BACKGROUND: Nitric oxide (NO) produced by the inducible isoform of NO synthase (iNOS or NOS2) has been implicated in the hypotension, organ failure, and death that complicate sepsis. To avoid the confounding effects and limitations of iNOS inhibitors, we used iNOS gene "knockout" mice to examine the effect of inducible NO production in a model of polymicrobial abdominal sepsis treated with antibiotics. We hypothesized that iNOS gene deficiency would significantly alter outcome. METHODS: C57BL6 wild-type (control) and congenic iNOS knockout mice were studied concurrently. Under halothane anesthesia, the ceca were ligated with 4-0 silk suture and punctured twice with a 26-gauge needle (cecal ligation and puncture, CLP). Survival was followed for 7 days, after which necropsies were performed in surviving animals. In an accompanying study examining the acute effects of sepsis, organ injury at 18 hours after CLP as determined by histology and the degree of cell death by apoptosis were examined with the use of hematoxylin and eosin (H&E) and TUNEL staining and two-channel fluorescence activated cell sorter (FACS) analysis. RESULTS: Sham laparotomy produced no lethality in either knockout (n = 3) or wild-type (n = 3) animals. Compared with survival in controls (n = 20), survival after CLP in iNOS knockout mice (n = 21) was significantly decreased (P < .01 at 2 days, P = .080 at 7 days, Mantel Haenszel log-rank test). CLP-induced apoptotic cell death was significantly less in the thymus of iNOS knockout mice compared with wild-type mice. CONCLUSIONS: We conclude that iNOS gene function provides a survival benefit in septic mice and is associated with increased sepsis-induced thymocyte apoptosis. To our knowledge, this is the first survival study examining the effect of iNOS gene deficiency in a clinically relevant model of sepsis. PMID- 10455920 TI - Redox regulation of the rat hepatocyte iNOS promoter. AB - BACKGROUND: The purpose of this study was to define the redox sensitive cis acting transcriptional mechanisms that regulate inducible nitric oxide synthase (iNOS) promoter function in the hepatocyte. METHODS: Clonal deletion constructs of the rat hepatocyte iNOS promoter (Genbank X95629; 1845 base pair)-reporter plasmid were transiently transfected into HepG2 cells treated with IL-1 beta and IL-1 beta + hydrogen peroxide. RESULTS: A segment of the promoter upstream from nucleotide -1126 was associated with redox-sensitive augmentation of promoter activity. Site-directed mutagenesis of 2 antioxidant response elements (AREs) was combined with transfection analysis to demonstrate that mutation of the ARE at nt 1347 ablated oxidant stress-mediated activation of the iNOS promoter. CONCLUSIONS: This ARE conveys the redox-sensitive response of the rat iNOS promoter. Hepatocyte iNOS expression is a novel and, as yet, poorly described antioxidant mechanism that is cytokine and redox sensitive and that plays a pleuripotent regulatory role in hepatocellular function in the face of sepsis and shock. PMID- 10455919 TI - Antisense oligonucleotides to c-fos and c-jun inhibit intimal thickening in a rat vein graft model. AB - BACKGROUND: C-fos and c-jun are 2 immediate early genes that have been implicated in the stimulation of vascular smooth muscle cell proliferation and migration. In previous experiments in our laboratory with a rat vein graft model a 2- to 3-fold increase of messenger RNA of c-fos and c-jun were noted 1 hour after vein graft perfusion. Because c-fos and c-jun are up-regulated after the perfusion of vein grafts, the purpose of this study was to delineate the temporal expression of c fos and c-jun protein and to study the effect of antisense oligonucleotides (ASO) to c-fos and c-jun on intimal thickening observed in this model. METHODS: Sprague Dawley rats underwent bilateral interposition femoral artery grafts with use of the superficial epigastric vein, which was harvested from 15 minutes up to 2 weeks and analyzed by Western blot for Fos and Jun protein. Additional rats underwent bypasses and at the time of the procedure 1 graft was treated with a pluronic gel containing an ASO to c-fos, c-jun, or sense and the contralateral side was treated with pluronic gel only. The vein grafts were harvested 2 weeks after the procedure and perfusion fixed. After longitudinal sectioning, the intimal and total wall thicknesses were measured in the perianastamotic and midgraft regions by a morphometric digitizing microscope and the statistics were analyzed by a paired Student's t test. RESULTS: Protein analysis by Western blot showed that c-fos levels rose quickly within 2 hours and leveled at 6 hours 40 fold above basal levels after vein graft perfusion. Similarly, c-jun levels rose 10-fold above basal levels after 15 minutes and peaked at 2 hours 120-fold above basal levels. The treatment of the vein grafts with these ASOs resulted in a reduction of about 30% in the thickness of the intimal layer and the total wall thickness in both the perianastomotic and the midgraft regions, which was statistically significant different from control veins. CONCLUSION: These results indicate a possible therapeutic role for ASO to immediate early genes in the treatment of vein graft intimal hyperplasia. PMID- 10455921 TI - Interleukin 10 is not essential for survival or for modulating T-cell function after injury. AB - BACKGROUND: Interleukin 10 (IL-10) is thought to be protective in injury and sepsis. However, we recently reported that IL-10 antagonism can be beneficial after burn injury. This study used IL-10-deficient (IL-10 [-/-]) mice to further define the role of IL-10 after injury. METHODS: Wild-type (WT) C57BL/6 or IL-10 ( /-) mice were anesthetized, sham or burn injured, and immunized subcutaneously with a T-cell-dependent protein antigen. Ten days later antigen-specific serum antibody isotype formation was measured by enzyme-linked immunosorbent assay. In addition, antigen-stimulated splenic T-cell proliferation and cytokine production (interleukin 2, interferon gamma, and tumor necrosis factor-alpha) were measured. RESULTS: Burn-injured IL-10 (-/-) mice survival (80%) was equivalent to that of burn-injured WT mice (74%). An injury-dependent loss of T-helper 1 (Th1)-type antibody isotype (IgG2a) formation occurred in both WT and IL-10 (-/-) mice. In vitro studies indicated that burn injury caused reduced antigen-stimulated splenic T-cell proliferation and Th1-type (interleukin 2 and interferon gamma) cytokine production in WT and IL-10 (-/-) mice, whereas burn-injured IL-10 (-/-) mice produced high levels of antigen-stimulated tumor necrosis factor-alpha. CONCLUSIONS: IL-10 is not essential for survival after burn injury or for several injury-induced changes in adaptive immune function, including Th1-type antibody isotype formation, T-cell proliferation, and Th1-type cytokine production. PMID- 10455922 TI - Effects of FR167653 on pulmonary ischemia-reperfusion injury: administration timing. PMID- 10455923 TI - Energy metabolism and mitochondrial damage during pulmonary preservation. PMID- 10455924 TI - Cyclophosphamide 200 mg/kg lacks ability to induce pluripotent stem cell engraftment in mice. PMID- 10455925 TI - IL-4 production in IDDM-nonrecurrent pancreas-transplanted BB rats with donor derived NKR-P1+TCR alpha beta + (NKT) cells, but not in IDDM-recurrent BB rats. PMID- 10455926 TI - Attempts to reveal the mechanism of CD95-ligand-mediated inflammation. PMID- 10455927 TI - Effect of adenovirus-mediated gene transfer with CTLA4-Ig gene in organ transplantation. PMID- 10455928 TI - In vitro and in vivo effects of an anti-CD45 monoclonal antibody in the rat. PMID- 10455929 TI - Medical activities and states of preparedness of donor hospitals. PMID- 10455930 TI - Neurosurgeons request better relationships in the Japanese Organ Transplantation System. Organ Transplantation and Brain Death Investigation Committee of the Japan Neurosurgical Society. PMID- 10455931 TI - Overseas liver transplantation for Japanese patients. PMID- 10455932 TI - Indication and strategy for adult living related liver transplantation. PMID- 10455933 TI - Living-related donor liver transplantation in adults: experience at Shinshu University Hospital. AB - Several considerations and improved techniques for performing LRLT in adults have been discussed. Taking into account the worldwide shortage of cadaveric organ grafts, the significance of LRLT for adult patients will probably never diminish. Further study is needed to clarify the factors governing the outcome of adult-to adult LRLT. PMID- 10455934 TI - Liver transplantation for autoimmune hepatitis: rejection and recurrence. PMID- 10455935 TI - Timing of heart transplantation and posttransplant care: overseas experience. PMID- 10455936 TI - Predicting outcomes and management of candidates for heart transplantation. PMID- 10455937 TI - Management of young potential candidates for heart transplantation. PMID- 10455939 TI - Analysis of the effects of recipient age and pretransplant hemodialysis period on cadaveric kidney transplant outcome using non-heart-beating donors. PMID- 10455938 TI - Mechanisms of exercise response in the denervated heart after transplantation. PMID- 10455940 TI - The beneficial effects of FK 506 on living-related renal transplantation in presensitized recipients. PMID- 10455942 TI - Differences between cyclosporin A and tacrolimus in organ transplantation. AB - In addition to their effect on the TCR-mediated signal transduction, both CyA and tacrolimus affect a wide variety of other biological processes, including induction of cell exocytosis and side effects. Although calcineurin has been implicated directly or indirectly in many of these processes, it seems plausible that the mechanisms by which these two drugs exert their full immunosuppressive activity are not necessarily the same. The cellular and molecular networks under differential immunosuppression by these two drugs remain to be explored. However, considering that investigations in this area are still ongoing, the answers will undoubtedly be available in the not too distant future. PMID- 10455941 TI - Decrease of free radical level in organ perfusate by a novel water-soluble carbon sixty, hexa(sulfobutyl)fullerenes. PMID- 10455943 TI - Changes in quality of life and working capacity before and after kidney transplantation. PMID- 10455944 TI - Current status of organ transplantation in Korea. PMID- 10455945 TI - Organ transplant surgery and biology in Taiwan. PMID- 10455946 TI - Current status of organ transplantation in Japan. PMID- 10455947 TI - Cytokine induction owing to LVAD support in canine models. PMID- 10455948 TI - Our experience with mechanical circulatory support for end-stage heart failure. PMID- 10455949 TI - Left ventricular assist systems as a bridge to heart transplantation: the role and the strategy in Japan. PMID- 10455950 TI - Application of ventricular assist systems for end-stage cardiomyopathy patients as a bridge to heart transplant or recovery. PMID- 10455951 TI - Technical comparison of liver-pancreas procurement. PMID- 10455952 TI - Importance of machine perfusion flow in kidney preservation. PMID- 10455953 TI - Evaluation and procurement of donor heart and lung in multiple organ harvesting. PMID- 10455954 TI - Present and future of the Japan Organ Transplantation Network. PMID- 10455955 TI - Strategy in combined pancreas and kidney transplantation from non-heart-beating cadaver donors. PMID- 10455957 TI - Protective mechanism of preconditioning hypoxia attenuates apoptosis formation during renal ischemia/reperfusion phase. PMID- 10455956 TI - Elimination of hyperinsulinemia after pancreas transplantation in rats. PMID- 10455958 TI - Efficacy and current problems with a porcine hepatocyte-based bioartificial liver: light and shade. PMID- 10455959 TI - Development of a packed-bed type bioartificial liver: tissue engineering approach. PMID- 10455960 TI - E2F decoy suppresses E-selectin expression in murine cardiac allograft arteriopathy. PMID- 10455961 TI - Late multiglycosidorum tripterygium treatment ameliorates established graft coronary arteriosclerosis after heart transplantation in the rat. PMID- 10455962 TI - Estradiol effects on nitric oxide synthase expression in the rat aorta allograft. PMID- 10455964 TI - Status of bone banks in Japan. PMID- 10455963 TI - Increased bile duct complications and/or chronic rejection in crossmatch positive human liver allografts. PMID- 10455965 TI - Problems in utility and safety of otological allografts. PMID- 10455966 TI - Postoperative management following heart transplantation. AB - Advances in surgical techniques, postoperative care, and experience have led to improved outcome in heart transplant patients. Specifically, the use of corticosteroid-free immunosuppression has reduced the risk of infection. The use of pravastatin early after transplantation has led to a decrease in clinically severe rejection episodes, improvement in survival, and reduction in transplant coronary artery disease. Reduction in natural-killer-cell cytotoxicity in the pravastatin-treated patients suggests an adjunct immunosuppressive effect of pravastatin in those patients on CyA-based immunosuppression. Quality of life has also improved in the heart transplant recipient with cardiac rehabilitation demonstrating a beneficial role in the improvement of exercise capacity. Newer immunosuppressive agents and strategies continue to demonstrate benefit in improving survival and the quality of life of the heart transplant recipient. PMID- 10455967 TI - Problems in preservation of allogeneic heart values and vessels. PMID- 10455968 TI - Combined use of deoxyspergualin and FK 506 for temporary circulatory assist. PMID- 10455969 TI - Immunosuppressive effect of triptolide in vitro. PMID- 10455970 TI - A randomized multicenter study on kidney preservation comparing HTK with UW. PMID- 10455971 TI - Improved graft function using a new myocardial preservation solution: Celsior. Preliminary data from a randomized prospective study. PMID- 10455973 TI - Ischemia-reperfusion injury in experimental models of organ transplantation. PMID- 10455972 TI - Preservation solutions for transplantation. AB - Eurocollins has almost been abandoned because of the glucose disadvantage. UW is certainly the most used preservation solution for livers, kidneys, and pancreases with excellent clinical and experimental preservation data. UW can certainly be considered the current golden standard solution. However, the disadvantage of high viscosity, high price, uneasy handling of many 1-L bags, and the fact that the radical scavenger glutathion cannot be detected in the bags by chemical analysis (presumably due to diffusion) encourage competitors to produce new compounds with better cost to effect ratios. HTK has a firm place in cardiac preservation; by demonstration of equal safety and efficacy in preserving livers and kidneys, at least in the middle and lower range of cold ischemia time, HTK will be sued more frequently, particularly with the consideration of lower price and more easy handling aspects. The suggested high volume perfusion is not really necessary, calculation based on a total volume of 10 L for a multiorgan donor show significant cost reductions. Celsior is current only used for cardiac preservation. Beyond all aspects of conservation and preservation potencies of all these fluids, it must not be forgotten that cold ischemia itself is a risk factor for organ function. Therefore, cold ischemia time should be kept as short as possible. People are willing to accept 24 hours or more cold ischemia time in kidney transplantation because organ failure can be treated by dialysis. In other organs, where immediate organ function is essential, like in clinical heart transplantation, cold ischemia is hardly ever extended beyond 6 hours. Why are hearts and kidneys so different? Very likely, there is no difference, and the outstanding results in living unrelated kidney transplants is mostly due to short cold ischemia time. PMID- 10455974 TI - Preservation of the liver: is it possible to extend the time of storage? PMID- 10455975 TI - Delayed kidney function risk score: donor factors versus ischemia/reperfusion injury. PMID- 10455976 TI - Myocardial protection for transplantation. PMID- 10455977 TI - Risk factors for primary dysfunction after liver transplantation in the University of Wisconsin solution era. PMID- 10455978 TI - The state of renal preservation for transplantation in New York. PMID- 10455979 TI - The role of cytokines and antioxidant status in graft quality prediction. PMID- 10455980 TI - Preservation of the pancreas for transplantation. PMID- 10455981 TI - Use of non-heart-beating donors: preliminary experience with perfusion in situ. PMID- 10455982 TI - Isolation of hematopoietic stem cells from heparinized cadaveric multiple organ donors: potential clinical implications. PMID- 10455983 TI - Effects of lidocaine on rat pancreatic islet metabolism. PMID- 10455984 TI - Renal allograft rupture: a clinicopathologic study of 37 nephrectomy cases in a series of 628 consecutive renal transplants. PMID- 10455985 TI - Single-center analysis of the incidence of acute rejection in cadaveric renal allograft recipients induced with low-dose OKT3 with and without mycophenolate mofetil. PMID- 10455986 TI - Clinical study of insulin-like growth factor-I and compensatory renal growth in patients with donor nephrectomy. PMID- 10455987 TI - Tissue factor and tissue factor pathway inhibitor in kidney allograft recipients. PMID- 10455988 TI - Short-term reproducibility of total homocysteine determinations in stable renal transplant recipients. PMID- 10455989 TI - Gastric mucosal calcinosis in renal transplant patients. PMID- 10455990 TI - En bloc skin excision for upper extremity cutaneous malignancies in renal allograft patients. PMID- 10455991 TI - Bone mass and body composition in children with chronic cholestasis before and after liver transplantation. PMID- 10455992 TI - Ischemic colitis secondary to venous thrombosis following a previous orthotopic liver transplant. PMID- 10455993 TI - Autohepatocyte transplantation in a degradable collagen device in the bursa omentalis. PMID- 10455994 TI - TGF-beta 1 inhibits HLA-DR and beta 2-microglobulin expression in HeLa cells induced with r-IFN. PMID- 10455995 TI - Modulation of Kupffer cell activity by muramyl dipeptide ameliorates normothermic liver ischemia/reperfusion in rats. PMID- 10455996 TI - A welcome arterial variation for pancreas transplantation. PMID- 10455997 TI - Effect of cyclosporine on glucose tolerance in streptozotocin-diabetic rats: cyclosporine may deteriorate insulin sensitivity. PMID- 10455998 TI - Effect of cyclophosphamide pretreatment and culture on insulin secretion by isolated monkey pancreatic islets (Macaca radiata radiata). PMID- 10455999 TI - Acute myopathy induced by colchicine in a cyclosporine treated heart transplant recipient: possible role of the multidrug resistance transporter. PMID- 10456000 TI - Acute parvovirus infection in a heart transplant recipient. PMID- 10456001 TI - Neurologic complications following heart transplantation in the modern era: decreased incidence, but postoperative stroke remains prevalent. PMID- 10456002 TI - Interaction between lovastatin and cyclosporine A after heart and kidney transplantation. PMID- 10456003 TI - Dura mater transplantation: 25 years' clinical experience. PMID- 10456004 TI - The anti-factor D antibody, MAb 166-32, inhibits the alternative pathway of the human complement system. PMID- 10456005 TI - Time course of expression of ICAM-1 and LFA-1 in rat lung allografts. PMID- 10456007 TI - Attitudes of physicians and nursing staff members toward organ donation in an urban area of Germany. PMID- 10456006 TI - Beneficial effects of ginsenosides of stems and leaves on cardiac and coronary vascular functions after 12-hour rat heart preservation. PMID- 10456008 TI - The public's attitudes toward incentives for organ donation. PMID- 10456009 TI - Nurses dancing with wolves. PMID- 10456010 TI - Seven reasons to quit nursing. PMID- 10456014 TI - Palliating the pain of dying. PMID- 10456013 TI - A touchy subject. PMID- 10456012 TI - Fosphenytoin facts. PMID- 10456011 TI - What did you say? I can't quite understand your spoken order. PMID- 10456015 TI - How can we improve the way we perform our pain assessments to meet the needs of patients from diverse cultures? PMID- 10456016 TI - Letter from Macedonia. PMID- 10456017 TI - Summer hours. One small intervention changed a child's world. PMID- 10456018 TI - Mastering ABGs. The art of arterial blood gas measurement. PMID- 10456020 TI - Assessing the female reproductive system. PMID- 10456019 TI - A new approach to ABG interpretation. PMID- 10456021 TI - Myocarditis. PMID- 10456022 TI - Culturally competent drug administration. PMID- 10456023 TI - Emergency! Warfarin-induced necrosis. PMID- 10456024 TI - Safeguarding nurses and patients. PMID- 10456025 TI - Storing lifeblood. Cord blood stem cell banking. PMID- 10456026 TI - Coming of age with an ostomy. Life with a stoma may be especially difficult for teens. PMID- 10456027 TI - How can nurses combat mandatory overtime? PMID- 10456028 TI - Teaching and grieving. PMID- 10456029 TI - Transvection and chromosomal trans-interaction effects. PMID- 10456030 TI - First International Conference on Signal Transduction, Croatia. PMID- 10456031 TI - "The end of the beginning": molecular and cellular biology of gene therapy keystone. 14-20 January 1999. PMID- 10456032 TI - Review of AACR meeting: new research approaches in the prevention and cure of prostate cancer, 2-6 December 1998, Indian Wells, CA. PMID- 10456033 TI - Transcriptional deregulation in hereditary disorders and cancer: the 12th annual CABM symposium, October 21-22, 1998, Piscataway, NJ. AB - As can be seen from the above descriptions, the presentations at the CABM symposium provided an extraordinarily rich and diverse panorama of some of the most exciting science in current molecular biology. The presentations provided both a general overview and a detailed analysis of multiple biological systems, which despite their specific differences, also generated insights into important common themes. The success of any meeting is most appropriately measured by the kinds of questions that are provoked for future study, not merely by the recitation of past discoveries. In fact, the different presentations often raised highly similar questions for future study. At the most fundamental levels of transcriptional regulation, what are the signals that provide specificity of gene expression? What is the structural basis of specific protein-protein interactions, such as those between homeodomain proteins and beta-catenin-Lef1 interactions, and how are these determinants altered in transcriptional regulation in oncogenesis and in genetic diseases? How is specificity achieved in transcriptional repression, given that the fundamental biochemical reactions often involve modifications of relatively ubiquitous components such as histones? To what extent do changes in specificity of gene activation and repression or in chromosomal architecture mediate the kinds of developmental and oncogenic signals mediated through transcriptional regulators such as Myc, BCL6 and other basic helix-loop-helix proteins and the HMGI proteins? How do altered signaling pathways affect diseases of development and differentiation such as cardiovascular disorders and aging itself? What are the pathways that integrate extracellular signals and transcription during the process of organogenesis? How do fundamental cellular structures such as adhesion junctions, and the interactions of a cell with other cells and extracellular matrix impact on normal and abnormal development and on malignancy, and how do these levels of structure and function alter nuclear regulation of transcription and cell division? These are some of the recurrent questions raised in talk after talk at this symposium, questions that undoubtedly will provide the impetus for important discoveries that will be presented at future CABM symposia. PMID- 10456034 TI - Molecular determinants of sensitivity to antitumor agents. PMID- 10456035 TI - Regaining control of your practice. Physician empowerment through active 'followership'. PMID- 10456036 TI - What you'll need to give all-electronic presentations. Slide shows are giving way to video clips and sound. PMID- 10456037 TI - Antibiotic needed in quadriplegic patient? PMID- 10456038 TI - Medical and surgical treatment of Parkinson's disease. Strategies to slow symptom progression and improve quality of life. AB - Despite new medical and surgical therapy, mortality rates for Parkinson's disease remain unchanged. Nevertheless, symptom progression can be slowed and quality of life improved with current methods of treatment. Levodopa is the most effective drug for Parkinson's disease, but its long-term use is associated with significant motor complications. Dopamine agonists hold promise because of more sustained stimulation of dopamine receptors and possibly an antioxidant effect. Selegiline, amantadine, and anticholinergics are still used but must be employed with caution in the elderly. COMT inhibitors may be useful adjuncts to levodopa therapy but are plagued with serious adverse effects. Goals of therapy in patients less than 60 years of age include sparing levodopa therapy and providing neuroprotection. For patients 60 years and older, goals include maintaining cognitive status and treating symptoms. Surgical treatment includes globus pallidus internal-segment pallidotomy, deep brain stimulation, and fetal nigral transplantation. These hold promise for the future. PMID- 10456039 TI - Local anesthesia. Topical application, local infiltration, and field block. AB - Local anesthetics are warranted whenever a clinical procedure causes pain that could be eliminated by their use. Their effectiveness is influenced by many factors, particularly the choice of agent and the technique of administration. The authors discuss the clinical uses and advantages of common local anesthetics and describe three techniques used in the primary care setting: topical application, local infiltration, and field block. PMID- 10456040 TI - Weighing risks in abdominal aortic aneurysm. Best repaired in an elective, not an emergency, procedure. AB - Abdominal aortic aneurysms are believed to result from several factors, one probably being inflammation that leads to dilatation, plaque deposition, and degeneration of the arterial wall. Most of these aneurysms are asymptomatic, but abdominal or back pain, shock, and a pulsatile abdominal mass indicate rupture. Initial aneurysm size exceeding 5 cm (2 in.) in diameter and the presence of hypertension and COPD are important predictors of rupture. The overall operative mortality rate with elective repair of an abdominal aortic aneurysm has been reported to range from 0.9% to 5% at university medical centers, and it is only slightly higher at community hospitals. However, with a ruptured aneurysm and emergency repair, the mortality rate rises to about 75%. Several long-term studies using life-table methods have found that 5-year survival rates after aneurysm repair range from 49% to 84%. This rate is significantly better than the 5-year survival rate of patients who did not have an abdominal aortic aneurysm repaired. However, it is not as good as that of the normal age-matched population, probably because many patients with an aneurysm have concomitant coronary artery disease. PMID- 10456041 TI - 'Two feet-one hand' syndrome. A recurring infection with a peculiar connection. PMID- 10456043 TI - Food-borne disease in the 21st century. What challenges await us? AB - As the world shrinks and international commerce expands, food-borne illness is likely to become a major public health focus worldwide. Improved surveillance, community education, thorough understanding of the food production chain, use of HACCP strategies, and ionizing radiation are all important for reducing the risks. Primary care physicians play a crucial role in surveillance and early reporting, as well as in educating the public on the importance of basic food safety. PMID- 10456042 TI - Addressing emerging infections. The partnership between public health and primary care physicians. AB - In response to concerns about emerging infections, the Minnesota Department of Health, in conjunction with the Centers for Disease Control and Prevention, developed a model emerging infections program in 1995. The authors' experience with the program has demonstrated the key role clinicians have as partners with public health agencies in identifying and reporting disease cases, educating patients about infectious risks, and preventing emerging infections. This partnership is well illustrated by two examples from Minnesota: a recent outbreak of Neisseria meningitidis infection in a rural area and a laboratory surveillance study of invasive and drug-resistant Streptococcus pneumoniae infections in a metropolitan area. PMID- 10456044 TI - The medical impact of a bioterrorist attack. Is it all media hype or clearly a potential nightmare? AB - Even with the technologic sophistication available in the United States today, effectiveness in dealing with a bioterrorist event is limited. Current surveillance systems may be inadequate to detect attacks. Because the onset of illness after exposure to an agent is delayed, even the time and location of the attack may be vague. In addition, most of the medical community is unfamiliar with many of the high-threat diseases, so identification of the problem may be further delayed. Many of us who are involved in studying the many aspects of bioterrorism believe that it is not a question of if such an event will occur but rather when, as well as which agent will be used and how extensive the damage will be. Given the enormity of what is possible, we must prepare for a potential nightmare. PMID- 10456045 TI - The search for meaning in monoclonal protein. Is it multiple myeloma or monoclonal gammopathy of undetermined significance? AB - Multiple myeloma and MGUS are the two most common causes of monoclonal protein in serum or urine. The usually accepted diagnostic triad for multiple myeloma consists of a significant paraprotein in the serum or urine, more than 10% to 15% plasma cells in bone marrow, and the presence of bony lesions. Patients who meet the first two criteria but have no bony lesions, cytopenias, renal failure, or hypercalcemia may have smoldering myeloma, which often can be observed for a period of time before therapy is required. MGUS is characterized by a serum IgG monoclonal protein less than 3.5 g/dL or IgA paraprotein less than 2 g/dL, with no or only a small amount of protein in urine (Bence Jones protein < 1 g/24 hr). Less than 10% plasma cells are present in bone marrow, and patients have no lytic bony lesions, anemia, hypercalcemia, or renal insufficiency. Another important criterion for MGUS is stability of the monoclonal protein over time. Nonetheless, during long-term follow-up, an associated malignant process develops in about 30% of MGUS patients. Since none of the features defining MGUS is uniformly helpful in predicting the risk for malignant disease, patients should be followed up on a regular basis indefinitely. PMID- 10456046 TI - What is causing your patient's sexual dysfunction? Uncovering a connection with hypertension and antihypertensive therapy. AB - Arterial hypertension is associated with structural and functional alterations of the vessel walls. Because vascular endothelium plays a central role in the control of vascular tone, endothelial dysfunction can also cause certain types of erectile dysfunction. Erectile dysfunction is also a common side effect of certain drugs, including many antihypertensive agents. Physicians should be aware of potential sexual side effects of such drugs and take appropriate steps to alleviate persistent problems. Most important, physicians need to ask patients about sexual function and discuss the possibility of erectile dysfunction caused by antihypertensive therapy. Erectile dysfunction can be effectively treated in most patients, and many treatment options are available. Sildenafil therapy has revolutionized the management of this disorder, but this agent should be used with caution in certain patients taking nitrates. PMID- 10456047 TI - A rare cause of breast necrosis. Calciphylaxis. PMID- 10456048 TI - Intermittent prednisone adequate for arthritis and lupus? PMID- 10456049 TI - Can you spot the signs of elder mistreatment? AB - The incidence of elder mistreatment is expected to increase as the baby boom generation ages and more elderly people are living or receiving care at home. Despite increased awareness and reporting of other forms of domestic abuse and neglect, recognition and management of elder abuse lag far behind actual incidents. Drs Kruger and Moon describe the signs of mistreatment as well as the physician's role in reporting and management. PMID- 10456050 TI - Food-borne illness. PMID- 10456051 TI - Prediction and preparation, fundamental functions of the cerebellum. PMID- 10456052 TI - Impaired capacity of cerebellar patients to perceive and learn two-dimensional shapes based on kinesthetic cues. AB - This study addresses the issue of the role of the cerebellum in the processing of sensory information by determining the capability of cerebellar patients to acquire and use kinesthetic cues received via the active or passive tracing of an irregular shape while blindfolded. Patients with cerebellar lesions and age matched healthy controls were tested on four tasks: (1) learning to discriminate a reference shape from three others through the repeated tracing of the reference template; (2) reproducing the reference shape from memory by drawing blindfolded; (3) performing the same task with vision; and (4) visually recognizing the reference shape. The cues used to acquire and then to recognize the reference shape were generated under four conditions: (1) "active kinesthesia," in which cues were acquired by the blindfolded subject while actively tracing a reference template; (2) "passive kinesthesia," in which the tracing was performed while the hand was guided passively through the template; (3) "sequential vision," in which the shape was visualized by the serial exposure of small segments of its outline; and (4) "full vision," in which the entire shape was visualized. The sequential vision condition was employed to emulate the sequential way in which kinesthetic information is acquired while tracing the reference shape. The results demonstrate a substantial impairment of cerebellar patients in their capability to perceive two-dimensional irregular shapes based only on kinesthetic cues. There also is evidence that this deficit in part relates to a reduced capacity to integrate temporal sequences of sensory cues into a complete image useful for shape discrimination tasks or for reproducing the shape through drawing. Consequently, the cerebellum has an important role in this type of sensory information processing even when it is not directly associated with the execution of movements. PMID- 10456053 TI - Lateral cerebellar hemispheres actively support sensory acquisition and discrimination rather than motor control. AB - This study examined a new hypothesis proposing that the lateral cerebellum is not activated by motor control per se, as widely assumed, but is engaged during the acquisition and discrimination of tactile sensory information. This proposal derives from neurobiological studies of these regions of the rat cerebellum. Magnetic resonance imaging of the lateral cerebellar output nucleus (dentate) of humans during passive and active sensory tasks confirmed four a priori implications of this hypothesis. Dentate nuclei responded to cutaneous stimuli, even when there were no accompanying overt finger movements. Finger movements not associated with tactile sensory discrimination produced no dentate activation. Sensory discrimination with the fingers induced an increase in dentate activation, with or without finger movements. Finally, dentate activity was greatest when there was the most opportunity to modulate the acquisition of the sensory tactile data: when the discrimination involved the active repositioning of tactile sensory surface of the fingers. Furthermore, activity in cerebellar cortex was strongly correlated with observed dentate activity. This distinct four way pattern of effects strongly challenges other cerebellar theories. However, contrary to appearances, neither our hypothesis nor findings conflict with behavioral effects of cerebellar damage, neurophysiological data on animals performing motor tasks, or cerebellar contribution to nonmotor, perceptual, and cognitive tasks. PMID- 10456054 TI - Cerebellar guidance of premotor network development and sensorimotor learning. AB - Single unit and imaging studies have shown that the cerebellum is especially active during the acquisition phase of certain motor and cognitive tasks. These data are consistent with the hypothesis that particular sensorimotor procedures are acquired and stored in the cerebellar cortex and that this knowledge can then be exported to the cerebral cortex and premotor networks for more efficient execution. In this article we present a model to illustrate how the cerebellar cortex might guide the development of cortical-cerebellar network connections and how a similar mechanism operating in the adult could mediate the exportation of sensorimotor knowledge from the cerebellum to the motor cortex. The model consists of a three-layered recurrent network representing the cerebello thalamocortical-ponto-cerebellar limb premotor network. The cerebellar cortex is not explicitly modeled. Our simulations show that Hebbian learning combined with weight normalization allows the emergence of reciprocal and modular structure in the limb premotor network. Reciprocal connections allow activity to reverberate around specific loops. Modularity organizes the connections into specific channels. Furthermore, we show that cerebellar learning can be exported to motor cortex through these modular and reciprocal premotor circuits. In particular, we simulate developmental alignment of visuomotor relations and their realignment as a consequence of prism exposure. The exportation of sensorimotor knowledge from the cerebellum to the motor cortex may allow faster and more efficient execution of learned motor responses. PMID- 10456055 TI - Role of cerebellum in adaptive modification of reflex blinks. AB - We investigated the involvement of the cerebellar cortex in the adaptive modification of corneal reflex blinks and the regulation of normal trigeminal reflex blinks in rats. The ansiform Crus I region contained blink-related Purkinje cells that exhibited a complex spike 20.4 msec after a corneal stimulus and a burst of simple spike activity correlated with the termination of orbicularis oculi activity. This occurrence of the complex spike correlated with trigeminal sensory information associated with the blink-evoking stimulus, and the burst of simple spike activity correlated with sensory feedback about the occurrence of a blink. Inactivation of the inferior olive with lidocaine prevented all complex and significantly reduced simple spike modulation of blink related Purkinje cells, but did not alter orbicularis oculi activity evoked by corneal stimulation. In contrast, both acute and chronic lesions of the cerebellar cortex containing blink-related Purkinje cells blocked adaptive increases in orbicularis oculi activity of the lid ipsilateral but not contralateral to the lesion. These data are consistent with the hypothesis that the cerebellum is part of a trigeminal reflex blink circuit. Changes in trigeminal signals produce modifications of the cerebellar cortex, which in turn, reinforce or stabilize modifications of brainstem blink circuits. When the trigeminal system does not attempt to alter the magnitude of trigeminal reflex blinks, cerebellar input has little or no effect on reflex blinks. PMID- 10456056 TI - Single-unit evidence for eye-blink conditioning in cerebellar cortex is altered, but not eliminated, by interpositus nucleus lesions. AB - Many theories of motor learning explain learning-related changes in motor behavior in terms of plasticity in the cerebellar cortex. Empirical evidence, however, does not always appear to be consistent with such formulations. It is the anterior cerebellar interpositus nucleus (aINP) that seems to be essential for acquisition and retention of conditioned eye-blink responses under most circumstances and it has been therefore suggested that the aINP is the critical site of learning-related plasticity during eye-blink conditioning. Supporting this conclusion are studies demonstrating that multiple-unit conditioning-related neural activity patterns observed in many brain regions disappear after aINP lesion. The possibility that the cerebellar cortex may be involved in forming these patterns has not been assessed adequately, however. In the current study, trained rabbits received kainic acid lesions of the INP. After recovery, the animals underwent additional sessions of conditioning during which single-unit activity was recorded from the cerebellar cortex. Our results suggest that the aINP is not the sole site of plasticity during eye-blink conditioning, as a subset of the neurons recorded from lesioned animals demonstrated conditioning related firing patterns. The lesions did change the character of these firing patterns from those observed in saline controls, however, in ways that can be generally described as a loss of organization. The normal tendency for the population of cortical cells to change firing rate together, for instance, was significantly less noticeable in lesioned animals. These results suggest that the aINP may be involved in the production of important features of conditioned responding, such as system timing function, therefore suggesting the need for more models that incorporate aINP and brain stem feedback as integral to the production of organized neural and behavioral responses. PMID- 10456057 TI - Effect of varying the intensity and train frequency of forelimb and cerebellar mossy fiber conditioned stimuli on the latency of conditioned eye-blink responses in decerebrate ferrets. AB - To study the role of the mossy fiber afferents to the cerebellum in classical eye blink conditioning, in particular the timing of the conditioned responses, we compared the effects of varying a peripheral conditioned stimulus with the effects of corresponding variations of direct stimulation of the mossy fibers. In one set of experiments, decerebrate ferrets were trained in a Pavlovian eye-blink conditioning paradigm with electrical forelimb train stimulation as conditioned stimulus and electrical periorbital stimulation as the unconditioned stimulus. When stable conditioning had been achieved, the effect of increasing the intensity or frequency of the forelimb stimulation was tested. By increasing the intensity from 1 to 2 mA, or the train frequency from 50 to 100 Hz, an immediate decrease was induced in both the onset latency and the latency to peak of the conditioned response. If the conditioned stimulus intensity/frequency was maintained at the higher level, the response latencies gradually returned to preshift values. In a second set of experiments, the forelimb stimulation was replaced by direct train stimulation of the middle cerebellar peduncle as conditioned stimulus. Varying the frequency of the stimulus train between 50 and 100 Hz had effects that were almost identical to those obtained when using a forelimb conditioned stimulus. The functional meaning of the latency effect is discussed. It is also suggested that the results support the view that the conditioned stimulus is transmitted through the mossy fibers and that the mechanism for timing the conditioned response is situated in the cerebellum. PMID- 10456058 TI - Conditioned response timing and integration in the cerebellum. AB - Classical conditioning procedures instill knowledge about the temporal relationships between events. The unconditioned stimulus (US) is the event to be timed. The conditioned response (CR) is viewed as a prediction of the imminence of the US. Knowledge of the elapsed time between conditioned stimuli (CSs) and US delivery is expressed in the topological features of the CR. The peak amplitude of the CR coincides with the timing of the US. A simple connectionist network based on Sutton and Barto's Time Derivative (TD) Model of Pavlovian Reinforcement provides a mechanism that can account for and simulate CR timing in a variety of protocols. This article describes extensions of the model to predictive timing under temporal uncertainty. The model is expressed in terms of equations that operate in real time according to a competitive learning rule. The unfolding of time from the onsets and offsets of events such as CSs is represented by the propagation of activity along a sequence of time-tagged elements. The model can be aligned with anatomical circuits of the cerebellum and brain stem that are essential for learning and performance of conditioned eye-blink responses. PMID- 10456059 TI - A model of Pavlovian eyelid conditioning based on the synaptic organization of the cerebellum. AB - We present a model based on the synaptic and cellular organization of the cerebellum to derive a diverse range of phenomena observed in Pavlovian eyelid conditioning. These phenomena are addressed in terms of critical pathways and network properties, as well as the sites and rules for synaptic plasticity. The theory is based on four primary hypotheses: (1) Two cerebellar sites of plasticity are involved in conditioning: (a) bidirectional long-term depression/potentiation at granule cell synapses onto Purkinje cells (gr-->Pkj) in the cerebellar cortex and (b) bidirectional plasticity in the interpositus nucleus that is controlled by inhibitory inputs from Purkinje cells; (2) climbing fiber activity is regulated to an equilibrium level at which the net strength of gr-->Pkj synapses remains constant unless an unexpected unconditioned stimulus (US) is presented or an expected US is omitted; (3) a time-varying representation of the conditioned stimulus (CS) in the cerebellar cortex permits the temporal discrimination required for conditioned response timing; and (4) the ability of a particular segment of the CS to be represented consistently across trials varies as a function of time since CS onset. This variation in across-trials consistency is thought to contribute to the ISI function. The model suggests several empirically testable predictions, some of which have been tested recently. PMID- 10456060 TI - Local dendritic Ca2+ signaling induces cerebellar long-term depression. AB - The coordinated activity of large numbers of adjacent parallel fiber synapses elevate calcium concentration locally in small regions of Purkinje cell dendrites. Such activity has also been reported to produce long-term depression of parallel fiber synaptic transmission. We have examined the relationship between these two events by combining patch clamp measurements of parallel fiber synaptic transmission with confocal microscopic imaging of the local calcium signals. We find that patterns of parallel fiber activity capable of evoking long term depression invariably cause increases in Purkinje cell calcium concentration that are very spatially restricted. These results suggest that one function of the local dendritic calcium signals is to induce long-term depression of parallel fiber synapses. PMID- 10456061 TI - Absence of cerebellar long-term depression in mice lacking neuronal nitric oxide synthase. AB - Extensive pharmacological evidence suggests that nitric oxide (NO) is a crucial transmitter for cerebellar long-term depression (LTD), a long-lasting decrease in efficacy of the synapses from parallel fibers onto Purkinje neurons, triggered by coincident presynaptic activity and postsynaptic depolarization. We now show that LTD cannot be induced in Purkinje neurons under whole-cell patch clamp in cerebellar slices from young adult mice genetically lacking neuronal nitric oxide synthase (nNOS). This genetic evidence confirms the essentiality of NO and nNOS for LTD in young adult rodents. Surprisingly, LTD in cells from nNOS knockout mice cannot be rescued by photolytic uncaging of NO and cGMP inside Purkinje neurons, although such stimuli circumvent acute pharmacological inhibition of nNOS and soluble guanylate cyclase in normal rodents. Also slices from knockout mice show no deficit in cGMP elevation in response to exogenous NO. Therefore, prolonged absence of nNOS allows atrophy of the signaling pathway downstream of cGMP. PMID- 10456062 TI - Environmental signals modulate olfactory acuity, discrimination, and memory in Caenorhabditis elegans. AB - Caenorhabditis elegans uses a variety of attractive olfactory cues to detect food. We show here that the responses to olfactory cues are regulated in a dynamic way by behavioral context and the animal's previous experience. Prolonged exposure to an odorant leads to a decreased response to that odorant, a form of behavioral plasticity called olfactory adaptation. We show that starvation can increase the extent of olfactory adaptation to the odorant benzaldehyde; this effect of starvation persists for several hours after the animals have been returned to food. The effect of starvation is antagonized by exogenous serotonin, which induces many of the same behavioral responses in C. elegans as are induced by food. Starvation also inhibits recovery from adaptation to a different odorant, 2-methylpyrazine, thus enhancing olfactory memory. In addition to its effects on adaptation, starvation modulates olfactory discrimination in C. elegans; starved animals discriminate more classes of odorants than fed animals. Increased olfactory discrimination is also seen in the adaptation-defective mutant adp-1 (ky20). These various forms of behavioral plasticity enhance the ability of starved animals to respond to novel, potentially informative odorants. PMID- 10456063 TI - Association of visual objects and olfactory cues in Drosophila. AB - Context-dependent preferences in a choice between an upper and a lower visual object of otherwise identical appearance were recorded during stationary flight of the fruitfly, Drosophila melanogaster, in a flight simulator. The test animal was held in a fixed orientation at the center of a wing-beat processor that converts attempted turns into counter-rotations of a surrounding cylindrical panorama. This allowed the fly to maneuver the preferred object into the actual direction of flight. Single flies were trained to avoid a course toward the visual object that had been associated with the aversive odor benzaldehyde (BAL). Conditioned object avoidance was investigated in different treatment groups by collective evaluation of the scores from 80 long-lasting flights (> 1 hr). In addition to a significant cross-modal association, we found a striking long-term effect of transient exposure to BAL both in the embryonic and larval states. The preimaginal experience significantly increased the indifference to BAL in the adult flies. Disturbed vision does not account for this effect: Neither the perception nor the discrimination of the visual objects was significantly impaired in the investigated flies. Disturbed olfaction could explain the present results. Recently, however, preimaginal BAL uptake has been found to interfere directly with the retention of heat-shock-conditioned object avoidance. PMID- 10456064 TI - Memory consolidation in Drosophila operant visual learning. AB - A new conditioning procedure, developed for the operant learning paradigm at the flight simulator, leads to stronger learning scores in wild-type flies. This procedure produces mean learning indices up to LI = 0.52. The heat-avoidance behavior acquired during training is very difficult to extinguish during extinction. Memory decays quickly during the first 3 hr after training and still is measurable 48 hr later. Disruption experiments demonstrate that memory can be disrupted by cold anesthesia within the first approximately 20 min after training, whereas KCl- and CXM-feeding regimens abolish memory soon after training and 3 hr later, respectively. These results initially suggest at least three distinct memory phases involved in memory consolidation in Drosophila after operant conditioning. PMID- 10456065 TI - Vision affects mushroom bodies and central complex in Drosophila melanogaster. AB - The brain of Drosophila is structurally altered by sensory stimuli that the flies receive during their adult life. Size and fiber number of the mushroom bodies, central complex, and optic lobes are influenced by social, spatial, or olfactory cues. Recently, the optic lobes have been shown to depend on the light regime that flies experience. Structural plasticity in the brain is thought to be a correlate of functional adaptations and long-term memory. We therefore extend our investigation of volume changes to the calyces of the mushroom bodies and the central complex. We show that rearing flies in constant light for 4 days increases the volume of both structures by up to 15% compared to rearing them in total darkness. Much of this difference develops during the first day. The effect of light is not hormonally mediated, as monocularly deprived flies develop a smaller ipsilateral calyx. Mutant analysis suggests that light generates its effects through known visual pathways. In contrast to the optic lobes, in the calyx and central complex structural changes can be linked to cAMP signaling, as in the mutants dunce1 and amnesiac1 no volume differences are observed. Surprisingly, the mutant rutabaga1 shows a prominent light-dependent volume increase in the calyx and central complex, dissociating structural from behavioral plasticity. In complete darkness wild-type flies grow larger calyces under crowded conditions in their normal culture vials than if kept in small groups on fresh food. This stimulating effect of crowding is not observed in any of the cAMP mutants, including rutabaga1. PMID- 10456066 TI - Brief theta-burst stimulation induces a transcription-dependent late phase of LTP requiring cAMP in area CA1 of the mouse hippocampus. AB - Memory storage in the mammalian brain can be divided into a short-term phase that is independent of new protein synthesis and a long-term phase that requires synthesis of new RNA and proteins. A cellular model for these two phases has emerged from studies of long-term potentiation (LTP) in the three major excitatory synaptic pathways in the hippocampus. One especially effective protocol for inducing robust and persistent LTP is "theta-burst" stimulation, which is designed to mimic the firing patterns of hippocampal neurons recorded during exploratory behavior in intact awake animals. Unlike LTP induced by non theta tetanization regimens, little is known about the biochemical mechanisms underlying theta-burst LTP in the hippocampus. In the present study, we examined theta-burst LTP in the Schaffer collateral pathway. We found that 3 sec of theta burst stimulation induced a robust and persistent potentiation (theta L-LTP) in mouse hippocampal slices. This theta L-LTP was dependent on NMDA receptor activation. The initial or early phase of theta-LTP did not require either protein or RNA synthesis and was independent of cAMP-dependent protein kinase (PKA) activation. In contrast, the late phase of theta-LTP required synthesis of proteins and RNA and was blocked by inhibitors of PKA. Prior induction of theta LTP also occluded the potentiation elicited by chemical activation of PKA. Our results show that, like non-theta LTP, theta-induced LTP in area CA1 of the mouse hippocampus also involves transcription, translation, and PKA and suggest that cAMP-mediated gene transcription may be a common mechanism responsible for the late phases of LTP induced by both theta and non-theta patterns of stimulation. PMID- 10456067 TI - Cellular mechanisms of visual cortical plasticity: a game of cat and mouse. PMID- 10456068 TI - Differential patterns of c-fos mRNA expression in amygdala during successive stages of odor discrimination learning. AB - Expression of the activity-dependent gene c-fos was used to assess relative levels of neuronal activation in the amygdala and related structures of rats at different stages of odor discrimination learning. In situ hybridization was used to evaluate c-fos mRNA content within the amygdalar subdivisions, the bed nucleus of the stria terminalis, and the hippocampus. After initial exploration of the test apparatus, c-fos mRNA levels were increased in the medial and, to lesser extent, basolateral subdivisions and remained low in the central division. The balance of amygdala to hippocampal labeling favored hippocampus. Rats engaged in familiar nose-poke responses had comparably elevated labeling in the medial and basolateral divisions and low labeling densities in the central division. The ratio of hippocampal to amygdala labeling was at control levels. Rats required to switch from ad libitum responding to cued responding to odors had high basolateral to medial labeling ratios. This was in marked contrast to the medial dominance found in control and exploration rats. Hybridization was substantially more dense in basolateral amygdala than in hippocampal CA1; this imbalance was unique to the group required to form first associations between odors and rewards. Rats performing an overtrained odor discrimination had the least differentiation between amygdalar subdivisions of any behavioral group. The hippocampus-to-amygdala labeling ratio favored hippocampus and was nearly identical to the ratio in exploration rats. These results demonstrate that the balance of activity within the between limbic structures shifts according to behavioral demands. It is suggested that the balances reflect the availability of pertinent afferent cues, interactions between hippocampus and the extended amygdala, and relative levels of activity in the diffuse projections to the limbic system. PMID- 10456069 TI - Reaction time distributions across normal forgetting: searching for markers of memory consolidation. AB - Volunteers studied pictures of objects and were then tested for yes/no recognition at 10 min and 1 week after learning (experiment 1), or at 10 min and 4 months after learning (experiment 2). Because the gradual consolidation of long term memory is thought to occur across this time scale (weeks and months), the reaction time distributions of successfully retrieved items were analyzed in an attempt to detect markers of consolidation. At each retention interval, reaction times for items retrieved successfully were well fit by a model that assumed a single underlying distribution. No evidence for a bimodal distribution of reaction times was observed. Furthermore, there was no evidence that some small subset of items was actually retrieved faster after a long retention interval than after a short interval. The results are consistent with the idea that consolidation works not to increase memory trace strength but to change the nature of memory storage. This process occurs during the course of normal forgetting and may not be observable in the behavior of normal memory. PMID- 10456070 TI - Impaired declarative memory for emotional material following bilateral amygdala damage in humans. AB - Everyday experience suggests that highly emotional events are often the most memorable, an observation supported by psychological and pharmacological studies in humans. Although studies in animals have shown that nondeclarative emotional memory (behaviors associated with emotional situations) may be impaired by lesions of the amygdala, little is known about the neural underpinnings of emotional memory in humans, especially in regard to declarative memory (memory for facts that can be assessed verbally). We investigated the declarative memory of two rare patients with selective bilateral amygdala damage. Both subjects showed impairments in long-term declarative memory for emotionally arousing material. The data support the hypothesis that the human amygdala normally enhances acquisition of declarative knowledge regarding emotionally arousing stimuli. PMID- 10456071 TI - Intact enhancement of declarative memory for emotional material in amnesia. AB - Emotional arousal has been demonstrated to enhance declarative memory (conscious recollection) in humans in both naturalistic and experimental studies. Here, we examined this effect in amnesia. Amnesic patients and controls viewed a slide presentation while listening to an accompanying emotionally arousing story. In both groups, recognition memory was enhanced for the emotionally arousing story elements. The magnitude of the enhancement was proportional for both amnesic patients and controls. Emotional reactions to the story were also equivalent. The results suggest that the enhancement of declarative memory associated with emotional arousal is intact in amnesia. Together with findings from patients with bilateral amygdala lesions, the results indicate that the amygdala is responsible for the enhancement effect. PMID- 10456072 TI - A biochemist's view of long-term potentiation. AB - This review surveys the molecular mechanisms of long-term potentiation (LTP) from the point of view of a biochemist. On the basis of available data, LTP in area CA1 of the hippocampus is divided into three phases--initial, early, and late- and the mechanisms contributing to the induction and expression of each phase are examined. We focus on evidence for the involvement of various second messengers and their effectors as well as the biochemical strategies employed in each phase to convert a transient signal into a lasting change in the neuron. We also consider, from a biochemical perspective, the implications of a multiphase model for LTP. PMID- 10456073 TI - c-Fos induction in the rat nucleus of the solitary tract correlates with the retention and forgetting of a conditioned taste aversion. AB - Recently, we have described a potential neuronal correlate of the behavioral expression of a conditioned taste aversion (CTA) against sucrose at the level of c-Fos expression. Intraoral infusions of sucrose induce c-Fos-like immunoreactivity (c-FLI) in the intermediate nucleus of the solitary tract (iNTS) after a CTA has been acquired for sucrose. Sucrose infusions do not induce c-FLI in the iNTS of unconditioned rats or in conditioned rats after extinction of the CTA. Here, we describe persistence of altered responsiveness of the iNTS in rats with CTAs against sucrose by intraorally infusing sucrose 2 days, 3 months, or 6 months after acquisition of the CTA. Sucrose infusions induced c-FLI in the iNTS 6 months after conditioning. The behavioral expression of the CTA was attenuated at 6 months but not at 3 months; the number of c-FLI positive cells in the iNTS was proportional to the magnitude of the expression of the CTA. This evidence strengthens our hypothesis that c-FLI in the iNTS is a neuronal correlate of the expression of a CTA. PMID- 10456074 TI - Transient expression of c-Fos in rat amygdala during training is required for encoding conditioned taste aversion memory. AB - Local microinjection into rat amygdala of phosphorothioate modified oligodeoxynucleotides (ODNs) antisense to c-fos several hours before conditioned taste aversion (CTA) training impaired taste aversion memory tested 3-5 days after conditioning. In contrast, injection of the antisense ODNs several days before training, before testing, or into the basal ganglia, or injection of c-fos sense ODNs, had no effect on CTA memory. Inhibition of translation by local microinjection of anisomycin into the amygdala shortly before as well as during CTA training, but not several days before training or shortly before testing, also impaired CTA memory. We conclude that translation in general, and c-Fos translation in particular, in the amygdala during or immediately after CTA training is essential for encoding taste aversion memory. PMID- 10456075 TI - Examination of the role of cGMP in long-term potentiation in the CA1 region of the hippocampus. AB - The mechanisms underlying the generation of NMDA receptor-dependent LTP in the CA1 region of the hippocampus continue to receive a great deal of attention because of the postulated importance of LTP as a synaptic mechanism for learning and memory. It is well accepted that the initial induction of LTP occurs in the postsynaptic cell, but the site of expression remains controversial. One prominent hypothesis is that LTP involves the release of one or more retrograde messengers that act on the presynaptic terminal to enhance transmitter release. Recently, evidence has been presented that retrograde messengers function to activate presynaptic guanylyl cyclase and that the resulting rise in presynaptic cGMP levels, when accompanied by presynaptic activity, is responsible for generating an early component of LTP. We have tested this hypothesis by examining whether synaptic strength is increased by coupling tetanic stimulation with application of a membrane-permeable analog of cGMP. The experiments were done in the presence of an NMDA receptor antagonist to block postsynaptic induction mechanisms. Under a variety of experimental conditions, this manipulation failed to generate LTP, suggesting that an increase in cGMP levels accompanied by presynaptic activity is not sufficient to generate LTP in the CA1 region of the hippocampus. PMID- 10456076 TI - Conditioned visual flight orientation in Drosophila: dependence on age, practice, and diet. AB - Orientation preferences for visual patterns can be conditioned in tethered flies (Drosophila melanogaster) at the flight simulator. In a reversal conditioning procedure using heat as reinforcement, flies can be trained to successively prefer different flight orientations with respect to the patterns. As in many learning paradigms, conditioned responses are highly variable. Although during training most flies reliably avoid the heat and the corresponding flight orientations, in subsequent learning tests without heat some show no consistent preference for the permissive orientations. We have started to investigate the interindividual differences in learning performance and describe here three significant variables: the age of the animals, their experience in the flight simulator prior to the experiment, and the composition of the fly food. Flies learn more reliably at 3-4 days than at 1-2 days of age but learning indices do not increase further in even older flies. Learning is improved if flies are allowed to become familiar with the flight simulator before the start of the conditioning procedure. Most important, poor nutrition causes complete amnesia within three or four generations. The reverse shift from poor to nutritious food restores learning ability with an even longer delay. PMID- 10456077 TI - Learning from LTP: a comment on recent attempts to identify cellular and molecular mechanisms of memory. AB - In recent years disappointing evidence has emerged regarding all main lines of evidence supporting connections between long-term potentiation (LTP) and memory. The history of this research, including studies focusing on synaptic alterations following learning experience, on saturation of LTP, and on pharmacological and genetic manipulations of LTP, are discussed briefly and interpreted in light of their observed and inherent limitations. Other approaches, aimed at showing a continuity of plasticity from molecular to synaptic to circuit and systems levels of analysis, are highlighted as potentially more compelling future directions for this research. PMID- 10456078 TI - Long-lasting forms of synaptic potentiation in the mammalian hippocampus. PMID- 10456079 TI - Properties of LTP induction in the CA3 region of the primate hippocampus. AB - Activity-dependent changes in synaptic strength, such as long-term potentiation (LTP), have been proposed to underlie memory storage in the brains of all mammals, including humans. However, most forms of synaptic plasticity, including LTP, are studied almost exclusively in rodents and related species. Thus, the hypothesis that LTP is important in human memory relies on the assumption that LTP is similar in the primate and rodent brains. We have begun to test this hypothesis by studying the properties and mechanisms of LTP induction in area CA3 of hippocampal slices from cynomolgus monkeys. We have found that LTP can be induced reliably at both mossy fiber-CA3 and collateral/associational-CA3 synapses in the primate brain, and that the properties of LTP induction at these synapses are similar to what we and others have observed in experiments using hippocampal slices from rodents. Also, we have investigated the role of opioids in mossy fiber synaptic transmission and LTP and have found no effect of the opioid antagonist naloxone nor the opioid agonist dynorphin on mossy fiber synaptic transmission or potentiation. These data suggest that LTP in the primate and rat brains has a similar induction mechanism and, thus, that the rodent is a useful animal model in which to study synaptic modification such as LTP. PMID- 10456080 TI - Proactive and retrograde effects on LTP produced by theta pulse stimulation: mechanisms and characteristics of LTP reversal in vitro. AB - Previous studies have established that (1) a 1-min episode of theta pulse stimulation (TPS) is sufficient to reverse potentiation during the early phases of LTP in area CA1 without causing depression when administered to nonpotentiated pathways; (2) the magnitude of depotentiation is inversely related to the delay between LTP induction and reversal attempts; and (3) pharmacological facilitation of AMPA receptor-mediated currents significantly enhances the strength of the reversal mechanism. The present experiments confirm and extend these results by showing that the depotentiating action of TPS on prior LTP is antagonized by inhibitors of protein phosphatases and adenosine A1 receptors but is not affected by NMDA receptor blockade, and, moreover, that TPS interferes with subsequent LTP induction by triggering an inhibitory mechanism that is active for a few minutes and is blocked by phosphatase inhibition. The possible implications of these results are discussed. PMID- 10456081 TI - Long-term potentiation induces synaptic plasticity at nontetanized adjacent synapses. AB - The two forms of long-term potentiation (LTP) and one form of long-term depression (LTD) in hippocampal area CA1 are induced by different afferent tetani: LTD is induced by a 3- to 5-Hz tetanus, whereas higher frequencies are necessary for LTP mediated by NMDA receptors (25- to 50-Hz tetanus) and LTP mediated by voltage-dependent calcium channels (200-Hz tetanus). It has been suggested that the three forms are induced by graded increases in postsynaptic calcium, with LTD being induced at the lowest calcium concentration. We hypothesized that synapses near a site of LTP induction would elicit LTD owing to diffusion of calcium or its binding proteins. This was tested using a fixed multistimulating electrode array spanning stratum radiatum in area CA1 of rat hippocampal slices. The tetanized sites all displayed LTP. Nontetanized sites displayed LTD following a 50-Hz tetanus, whereas LTD was often seen following a 200-Hz tetanus. In most instances pEPSP and population spike responses were similar; however, EPSP/spike dissociations (LTD of EPSP, LTP of spike) were seen following activation of NMDA receptors by 50-Hz and 200-Hz tetani. The results are discussed with respect to mechanisms of action and functional significance. PMID- 10456082 TI - Differential expression of short-term potentiation by AMPA and NMDA receptors in dentate gyrus. AB - Both alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and N-methyl-D aspartate (NMDA) glutamatergic receptor subtypes in hippocampus have been shown to express long-term potentiation (LTP), a form of synaptic modification believed to be involved in memory formation. Because of their postsynaptic localization, any differential expression of LTP by the two receptor subtypes would strongly support the existence of a postsynaptic mechanism of LTP expression. In this study, electrophysiological recordings from dentate granule cells were used to compare the potentiation of AMPA and NMDA receptor-mediated responses occurring during the initial phase of LTP, typically identified as STP. Results revealed that high-frequency stimulation (HFS) of perforant path afferents induces a robust STP of both AMPA and NMDA receptor-mediated components of granule cell EPSPs (referred to as AMPA STP and NMDA STP, respectively). Although STP for both receptor subtypes decayed to an asymptotic, steady-state level of LTP and could be induced repetitively, there were substantial differences in several aspects of AMPA and NMDA STP dynamics. STP of the AMPA receptor reached its peak magnitude approximately 30 sec after HFS and decayed with a time constant of approximately 6 min. In contrast, peak magnitude of NMDA STP always appeared immediately after HFS and decayed with a time constant of only 1 min. Single-pulse stimulation of perforant path afferents paired with postsynaptic depolarization also induced LTP of both AMPA and NMDA components. When this induction paradigm was used, however, only the AMPA component showed significant STP. Our results demonstrate that AMPA and NMDA receptors exhibit markedly different degrees of activity-dependent, short-term modifiability, with the possibility that STP of the NMDA receptor reflects primarily post-tetanic potentiation (PTP). In addition, our results strongly suggest that the mechanisms underlying STP of the AMPA receptor are postsynaptic in origin. PMID- 10456083 TI - Functional integrity of NMDA-dependent LTP induction mechanisms across the lifespan of F-344 rats. AB - Previous studies have reported a lack of an age effect in the induction of long term potentiation (LTP) at CA1 synapses, using robust (supramaximal) stimulation parameters, but an apparent age effect on the induction threshold of LTP using less robust stimulation, in the perithreshold region. These findings have led to the suggestion that old animals may experience an alteration either in the efficacy of activation of N-methyl-D-aspartate (NMDA) receptors or in the metabolic processes subsequent to NMDA receptor activation that lead to LTP expression. An alternative explanation for the apparent threshold change in old animals is that, because of the known reduction of the intracellularly recorded, compound EPSP magnitude in old rats, equivalent electrical stimulation results in a smaller effective depolarization of the postsynaptic cells and a consequently less effective activation of NMDA receptors, which are otherwise functionally normal. To distinguish between these two hypotheses, weak orthodromic stimulation was paired with intracellularly applied current pulses, thus holding constant the degree of postsynaptic depolarization. No differences in LTP induction threshold or magnitude were observed in a large sample of rats from three age groups. It is concluded that the NMDA receptor mechanisms and associated biochemical processes leading to LTP induction are not altered in aged F-344 rats. The reduced compound EPSP in old animals was reconfirmed in the present study, and a significant correlation was found in old rats between the magnitude of the EPSP at a fixed stimulus level and their performance on a spatial memory task. PMID- 10456084 TI - Deficiency in induction but not expression of LTP in hippocampal slices from young rats. AB - In this study we examine developmental changes between postnatal day (PND) 4 and 14 in synaptic transmission and plasticity in the CA1 region of hippocampal slices. We confirm previous results that tetanus-induced long-term potentiation (LTP) in field recordings is diminished in slices from younger animals. LTP in whole-cell current-clamp recordings is also diminished in younger animals. However, robust LTP can be induced in young animals if sufficient postsynaptic depolarization is provided during LTP induction. Furthermore, we find differences in synaptic transmission between PND 4 and 14, suggesting that the depolarization during tetanic stimulation in young tissue is ineffective to produce LTP. These results indicate that the smaller potentiation in field recordings in slices from younger animals is attributable to insufficient postsynaptic depolarization during LTP induction rather than a defect in expression mechanisms. PMID- 10456085 TI - Fluctuations in intracellular calcium responses to action potentials in single en passage presynaptic boutons of layer V neurons in neocortical slices. AB - The release of neurotransmitter from a nerve terminal on invasion by an action potential shows large trial-to-trial fluctuations. The factors contributing to this variability have not been elucidated clearly. Here, simultaneous patch-clamp and optical measurements from layer V neocortical pyramidal neurons have been used to assess the extent to which the fluctuations in transmitter release may be caused by variability in calcium rise in presynaptic structures. Boutons on axon collaterals were visualized and increases in intracellular calcium, assessed by Fura-2, were observed in response to single action potentials. In some boutons, calcium responses showed trial-to-trial variability and occasional apparent failures despite the faithful conduction of the action potential. These results suggest that a factor contributing to the fluctuation in transmitter release may be the variability with which depolarization of a presynaptic bouton produces an increase in intrabouton calcium. PMID- 10456086 TI - The role of dendritic action potentials and Ca2+ influx in the induction of homosynaptic long-term depression in hippocampal CA1 pyramidal neurons. AB - Long-term depression (LTD) of synaptic efficacy at CA1 synapses is believed to be a Ca(2+)-dependent process. We used high-speed fluorescence imaging and patch clamp techniques to quantify the spatial distribution of changes in intracellular Ca2+ accompanying the induction of LTD at Schaffer collateral synapses in CA1 pyramidal neurons. Low-frequency stimulation (3 Hz), which was subthreshold for action potentials, produced small changes in [Ca2+]i and failed to elicit LTD. Increasing the stimulus strength so that action potentials were generated produced both robust LTD and increases in [Ca2+]i. Back-propagating action potentials at 3 Hz in the absence of synaptic stimulation also produced increases in [Ca2+]i, but failed to induce LTD. When subthreshold synaptic stimulation was paired with back-propagating action potentials, however, large increases in [Ca2+]i were observed and robust LTD was induced. The LTD was blocked by the N methyl-D-aspartate receptor (NMDAr) antagonist APV, and stimulus-induced increases in [Ca2+]i were reduced throughout the neuron under these conditions. The LTD was also dependent on Ca2+ influx via voltage-gated Ca2+ channels (VGCCs), because LTD was severely attenuated or blocked by both nimodipine and Ni2+. These findings suggest that back-propagating action potentials can exert a powerful control over the induction of LTD and that both VGCCs and NMDArs are involved in the induction of this form of plasticity. PMID- 10456087 TI - The balance between postsynaptic Ca(2+)-dependent protein kinase and phosphatase activities controlling synaptic strength. AB - The activities of protein kinases and phosphatases are believed to regulate neuronal activity and synaptic plasticity in brain. Numerous in vivo and in vitro studies have shown that synaptic strength appears stable under basal conditions and during long-term potentiation (LTP) expression. This may reflect a balance between protein kinase and phosphatase activities. To provide experimental evidence for this hypothesis, and based on our knowledge that Ca2+/CaM activates protein kinases and phosphatases and that postsynaptic Ca2+/CaM signal pathways play important roles in synaptic plasticity, we examined the contribution of postsynaptic Ca(2+)-dependent protein kinases and calcineurin (CaN) in regulating synaptic strength. We show that inhibiting postsynaptic Ca2+/CaM-dependent protein kinase II (CaM-KII) and Ca2+/phospholipitidyserine-dependent protein kinase (PKC) in hippocampal CA1 neurons attenuates significantly the expression of LTP, but not basal synaptic transmission. On the other hand, the inhibition of postsynaptic CaN enhances synaptic transmission at potentiated and naive synapses, and increases significantly the magnitude of synaptic potentiation during the induction phase of LTP. These results indicate that postsynaptic CaM KII and PKC activities are essential for maintaining LTP expression, but CaN activity limits synaptic strength at stable levels during both basal and potentiated synaptic transmission; that is, the dynamic balance between protein phosphorylation and dephosphorylation that sets physiological synaptic strength is dominated by CaN activity. PMID- 10456088 TI - Phorbol ester effects at hippocampal synapses act independently of the gamma isoform of PKC. AB - Ca2+/phospholipid-dependent protein kinase has long been thought to play an important role in modulating synaptic efficacy. It has been shown previously that mice lacking the brain-specific gamma subtype of PKC display abnormal long-term potentiation (LTP), whereas ordinary synaptic transmission is unaffected by the mutation. We now examine the effects of phorbol esters, which are nonselective activators of PKC, on synaptic modulation in these mutant mice. In wild-type mice, phorbol esters produce marked enhancement of synaptic transmission that is largely presynaptic in origin, an effect that has been thought to share mechanisms with LTP. In mutant mice, phorbol ester-mediated potentiation is normal despite the absence of the major PKC isoform. As in wild-type mice, this synaptic enhancement is at least partly attributable to presynaptic changes. Our results demonstrate that the gamma isotype of PKC is not essential for phorbol ester-mediated synaptic facilitation, and place limitations on the possible roles of PKC in LTP. PMID- 10456089 TI - Determinants of BDNF-induced hippocampal synaptic plasticity: role of the Trk B receptor and the kinetics of neurotrophin delivery. AB - The neurotrophins are a class of signaling molecules known for their growth and survival-promoting activities during neuronal development. Recent studies suggest that the neurotrophins, including brain-derived neurotrophic factor (BDNF), can also dramatically influence synaptic transmission in the adult hippocampus. The experiments described in this paper indicate that ability of BDNF to potentiate synaptic transmission in the hippocampus relies on functional Trk B receptors. Moreover, the rate at which BDNF is applied to hippocampal synapses is also a potent determinant of whether synaptic potentiation will result. Hippocampal slices perfused with BDNF at a very slow flow rate (e.g., < or = 25 ml/hr) did not show synaptic potentiation. Increasing the rate of BDNF application resulted in synaptic potentiation in which the magnitude and onset kinetics of the potentiation were determined by the rate of BDNF delivery. Immunocytochemical analysis of BDNF detected with confocal microscopy confirmed these electrophysiological observations, indicating that the penetration of BDNF into hippocampal slices is influenced dramatically by the perfusion rate. PMID- 10456090 TI - Alterations in the expression of specific glutamate receptor subunits following hippocampal LTP in vivo. AB - Quantitative in situ hybridization revealed that following the induction of hippocampal long-term potentiation (LTP) in the dentate gyrus of freely moving rats, specific increases in the expression of the NR2B subunit of the N-methyl-D aspartate (NMDA) receptor and mGluR1c, a short splice variant of the metabotropic glutamate receptors that are linked intracellularly to phospholipase C (PLC) and protein kinase C (PKC), were seen in the postsynaptic dentate granule cells. There were no changes in the expression of NR2A; NR2C and NR2D NMDA receptor subunits; or mGluR1a, mGluR1b, mGluR5a, and mGluR5b PLC-associated metabotropic receptors. The elevations in NR2B and mGluR1c mRNA were delayed, occurring days after LTP induction. NR2B expression was enhanced significantly by 48 hr after LTP but was starting to decrease toward basal levels by 96 hr. The transient increase in the expression of NR2B mirrored the increase in the expression of PKC sensitive isoforms of the NR1 subunits of the NMDA receptor we observed previously (Thomas et al. 1994a). The increase in mGluR1c expression was more persistent, showing a significant increase 96 hr after LTP. This study demonstrates that not only are there changes in the expression of individual glutamate receptor subunits but the increases in their expression occur days after the induction of LTP and may reflect so-called late-onset genes that may be important for the maintenance of LTP. PMID- 10456091 TI - Translational suppression of calpain blocks long-term potentiation. AB - Transfection with antisense oligonucleotides was used to reduce calpain 1 activity to approximately 50% of normal values in cultured hippocampal slices. This had no detectable effects on baseline synaptic responses but greatly reduced the incidence and magnitude of long-term potentiation induced with a theta-burst stimulation paradigm. These results suggest that activation of calpain by repetitive bursts of afferent activity, as shown to occur in prior studies, is an essential step in the production of stable increases in synaptic strength. PMID- 10456092 TI - Long-term potentiation is associated with new excitatory spine synapses on rat dentate granule cells. AB - To investigate possible morphological correlates to long-term potentiation (LTP), three-dimensional reconstruction of serial electron micrographs was employed. LTP was induced in the perforant path/dentate granule cell synapse in two rats. The surgically isolated contralateral side served as control, along with two untreated animals. Longitudinally sectioned and transversally sectioned dendrites were sampled from the middle fifth of the molecular layer and all visibly connected spines were identified. A mixed, unbalanced, nested variance component model was used to make a valid statistical comparison between the LTP and control groups. The spine density was higher in the experimental than in the control groups. The changes were statistically significant in both the longitudinal and transverse sample. In addition, spines with a divided stem and two heads (bifurcating spines) were seen at a higher frequency in the LTP material compared with the contralateral material. From a subset of dendrites all connected spines were reconstructed and detailed measurements of head, neck, and PSD dimensions were made. We failed to find significant differences following LTP on either of the dimensions measured. The results suggest that new spine synapses are formed following LTP, including some of the bifurcating type. PMID- 10456093 TI - Convergent but temporally separated inputs to lateral amygdala neurons from the auditory thalamus and auditory cortex use different postsynaptic receptors: in vivo intracellular and extracellular recordings in fear conditioning pathways. AB - The lateral nucleus of the amygdala (LA), a key component of the fear conditioning circuitry, receives a rapid but relatively impoverished auditory input from the auditory thalamus and a slower but richer input from the auditory cortex. We examined in urethane anesthetized rats whether individual cells in the LA receive convergent inputs from these two areas, and whether different postsynaptic receptors contribute to the temporally separated excitations over the two pathways. With both extracellular and intracellular recordings, individual cells could be activated by stimulation of each pathway. In extracellular recordings iontophoretic application of the N-methyl-D-aspartate (NMDA) receptor antagonist APV and the L-alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionate (AMPA) receptor antagonist CNQX demonstrated that synaptic transmission in both pathways depends on AMPA receptors, whereas transmission in the thalamic pathway also depends on the involvement of NMDA receptors. The involvement of NMDA receptors in synaptic activation of the LA from the thalamus but not the cortex was confirmed in intracellular recordings using systemic injections of the NMDA antagonist MK-801. The slow time course of NMDA currents could provide LA cells with a mechanism to integrate the inputs arriving rapidly from the thalamus and somewhat later from the cortex, thus allowing the LA to integrate signals in the two pathways during the acquisition and expression of conditioned fear reactions. PMID- 10456094 TI - Physiologically realistic formation of autoassociative memory in networks with theta/gamma oscillations: role of fast NMDA channels. AB - Recordings from brain regions involved in memory function show dual oscillations in which each cycle of a low-frequency theta oscillation (5-8 Hz) is subdivided into about seven subcycles by high frequency gamma oscillations (20-60 Hz). It has been proposed (Lisman and Idiart 1995) that such networks are a multiplexed short-term memory (STM) buffer that can actively maintain about seven memories, a capability of human STM. A memory is encoded by a subset of principal neurons that fire synchronously in a particular gamma subcycle. Firing is maintained by a membrane process intrinsic to each cell. We now extend this model by incorporating recurrent connections with modifiable synapses to store long-term memory (LTM). The repetition provided by STM gradually modifies synapses in a physiologically realistic way. Because different memories are active in different gamma subcycles, the formation of autoassociative LTM requires that synaptic modification depend on N-methyl-D-aspartate (NMDA) channels having a time constant of deactivation that is of the same order as the duration of a gamma subcycle (15-50 msec). Many types of NMDA channels have longer time constants (150 msec), as for instance those found in the hippocampus, but both fast and slow NMDA channels are present in cortex. This is the first proposal for the special role of these fast NMDA channels. The STM for novel items must depend on activity-dependent changes intrinsic to neurons rather than recurrent connections, which have not developed the required selectivity. Because these intrinsic mechanisms are not error-correcting, STM will become slowly corrupted by noise. This limits the accuracy with which LTM can become encoded after a single presentation. Accurate encoding of items in LTM can be achieved by multiple presentations, provided different memory items are presented in a varied interleaved order. Our results indicate that a limited memory-capacity STM model can be integrated in the same network with a high-capacity LTM model. PMID- 10456095 TI - Novel lists of 7 +/- 2 known items can be reliably stored in an oscillatory short term memory network: interaction with long-term memory. AB - This paper proposes a model for the short-term memory (STM) of unique lists of known items, as, for instance, a phone number. We show that the ability to accurately store such lists in STM depends strongly on interaction with the preexisting long-term memory (LTM) for individual items (e.g., digits). We have examined this interaction in computer simulations of a network based on physiologically realistic membrane conductances, synaptic plasticity processes, and brain oscillations. In the model, seven STMs can be kept active, each in a different gamma-frequency subcycle of a theta-frequency oscillation. Each STM is maintained and timed by an activity-dependent ramping process. LTM is stored by the strength of synapses in recurrent collaterals. The presence of preexisting LTM for an item greatly enhances the ability of the network to store an item in STM. Without LTM, the precise timing required to keep cells firing within a given gamma subcycle cannot be maintained and STM is gradually degraded. With LTM, timing errors can be corrected and the accuracy and order of items is maintained. This attractor property of STM storage is remarkable because it occurs even though there is no LTM that identifies which items are on the list or their order. Multiple known items can be stored in STM, even though their representation is overlapping. However, multiple, identical memories cannot be stored in STM, consistent with the psychophysical demonstration of repetition blindness. Our results indicate that meaningful computation (memory completion) can occur in the millisecond range during an individual gamma cycle. PMID- 10456096 TI - Theta/gamma networks with slow NMDA channels learn sequences and encode episodic memory: role of NMDA channels in recall. AB - This paper examines the role of slow N-methyl-D-aspartate (NMDA) channels (deactivation approximately 150 msec) in networks that multiplex different memories in different gamma subcycles of a low frequency theta oscillation. The NMDA channels are in the synapses of recurrent collaterals and govern synaptic modification in accord with known physiological properties. Because slow NMDA channels have a time constant that spans several gamma cycles, synaptic connections will form between cells that represent different memories. This enables brain structures that have slow NMDA channels to store heteroassociative sequence information in long-term memory. Recall of this stored sequence information can be initiated by presentation of initial elements of the sequence. The remaining sequence is then recalled at a rate of one memory every gamma cycle. A new role for the NMDA channel suggested by our finding is that recall at gamma frequency works well if slow NMDA channels provide the dominant component of the EPSP at the synapse of recurrent collaterals: The slow onset of these channels and their long duration allows the firing of one memory during one gamma cycle to trigger the next memory during the subsequent gamma cycle. An interesting feature of the readout mechanism is that the activation of a given memory is due to cumulative input from multiple previous memories in the stored sequence, not just the previous one. The network thus stores sequence information in a doubly redundant way: Activation of a memory depends on the strength of synaptic inputs from multiple cells of multiple previous memories. The cumulative property of sequence storage has support from the psychophysical literature. Cumulative learning also provides a solution to the disambiguation problem that occurs when different sequences have a region of overlap. In a final set of simulations, we show how coupling an autoassociative network to a heteroassociative network allows the storage of episodic memories (a unique sequence of briefly occurring known items). The autoassociative network (cortex) captures the sequence in short-term memory and provides the accurate, time compressed repetition required to drive synaptic modification in the heteroassociative network (hippocampus). This is the first mechanistically detailed model showing how known brain properties, including network oscillations, recurrent collaterals, AMPA channels, NMDA channel subtypes, the ADP, and the AHP can act together to accomplish memory storage and recall. PMID- 10456097 TI - Hippocampal CA3 region predicts memory sequences: accounting for the phase precession of place cells. AB - Hippocampal recordings show that different place cells fire at different phases during the same theta oscillation, probably at the peak of different gamma cycles. As the rat moves through the place field of a given cell, the phase of firing during the theta cycle advances progressively. In this paper we have sought to determine whether a recently developed model of hippocampal and cortical memory function can explain this phase advance and other properties of place cells. According to this physiologically based model, the CA3 network stores information about the sequence of places traversed during learning. Here we show that the phase advance can be understood if it is assumed that the hippocampus is in a recall mode that operates when the animal is already familiar with a path. In this mode, sensory information about the current position triggers recall of the upcoming 5-6 places (memories) in the path at a rate of one memory per gamma cycle. The model predicts that the average phase advance will be one gamma cycle per theta cycle, a value in reasonable agreement with the data. The model also correctly accounts for (1) the fact that the firing of a place cell occurs during approximately 7 theta cycles (on average) as the animal crosses the place field; (2) the observation that the phase of place cell firing depends more systematically on position than on time; and (3) the fact that traversal of an already familiar path produces further modifications (shifts the firing of a cell to an earlier position in the path). This later finding suggests that recall of previously stored information strengthens the memory of that information. In the model, this occurs because of a novel role of N-methyl-D aspartate channels in recall. The general success of the model provides support for the idea that the hippocampus stores sequence information and makes predictions of expected positions during gamma-frequency recall. PMID- 10456098 TI - Basal and apical synapses of CA1 pyramidal cells employ different LTP induction mechanisms. AB - Nitric oxide (NO) production has been widely reported to be required for the induction of long-term potentiation (LTP) in hippocampal CA1 cells. Of the two constitutive isoforms of NO synthase, the endothelial form (eNOS) has been implicated in the induction of LTP in these cells. The distribution of eNOS within CA1 cells is not uniform, however, being present in the cell bodies and apical dendrites but absent from the basal dendrites. Using extracellular and intracellular recording techniques, we demonstrate that LTP induction in stratum radiatum synapses (onto apical dendrites) is dependent on NO production, being attenuated by pretreatment with a NOS inhibitor. LTP induced in stratum oriens synapses (onto basal dendrites) is, however, resistant to NOS inhibitors. Both forms of LTP require the activation of N-methyl-D-aspartate (NMDA) receptors because induction of LTP in both stratum radiatum and stratum oriens is blocked by AP5. Thus, it appears that synapses onto apical and basal dendrites of CA1 cells use different cellular mechanisms of LTP induction. PMID- 10456099 TI - Training in the Morris water maze occludes the synergism between ACPD and arachidonic acid on glutamate release in synaptosomes prepared from rat hippocampus. AB - We report here that release of glutamate, inositol phospholipid metabolism, and protein kinase C (PKC) activity are increased in synaptosomes prepared from hippocampi of rats that had been trained in a spatial learning task. In hippocampi obtained from animals that were untrained, activation of the metabotropic glutamate receptor by the specific agonist trans-1-amino-cyclopentyl 1,3-dicarboxylate (ACPD) increased release of glutamate but only in the presence of a low concentration of arachidonic acid. A similar interaction between arachidonic acid and ACPD was observed on inositol phospholipid turnover and on PKC activity. However, the synergistic effect of arachidonic acid and ACPD on glutamate release was occluded in hippocampal synaptosomes prepared from trained rats. Occlusion of the effect on inositol phospholipid turnover and PKC activation was also observed. These data suggest that the molecular changes that underlie spatial learning may include activation of metabotropic glutamate receptors in the presence of arachidonic acid and that the interaction between arachidonic acid and ACPD triggers the presynaptic changes that accompany learning. PMID- 10456100 TI - Intact implicit memory for novel patterns in Alzheimer's disease. AB - Repetition priming is a kind of implicit memory (learning without awareness) that does not depend on the medial temporal-lobe system. For example, the amnesic patient H.M., who underwent bilateral medial temporal-lobe resection, shows intact priming with novel patterns, suggesting that perceptual priming with nonverbal material does not depend on areas critical for explicit memory. A logical candidate for the neural substrate that supports this kind of priming is the peristriate cortex, an area that is relatively spared in Alzheimer's disease (AD). We therefore predicted that AD subjects would be unimpaired on pattern priming. Subjects copied each of six target figures onto dot patterns. After performing a 3-min distractor task, they were given the same dot patterns (without lines) and asked to draw the first figure that came to mind by connecting the dots with straight lines. Subsequently, in a test of recognition (explicit) memory, subjects viewed each of the six patterns of dots that they had copied previously and were asked to indicate which of four possible completions corresponded to the figure that they had copied 3 min earlier. The AD and control groups achieved comparable priming scores, but AD subjects were significantly impaired in recognizing the patterns explicitly. Our finding of intact pattern priming in AD provides, for the first time, evidence that pattern priming depends on the peristriate cortex. PMID- 10456101 TI - Temporally graded retrograde amnesia following separate and combined lesions of the perirhinal cortex and fornix in the rat. AB - The involvement of the perirhinal cortex and the fornix in retrograde and anterograde amnesia in the rat was investigated in this experiment. Male Sprague Dawley rats were trained on a series of five visual discrimination problems at distinct time intervals prior to receiving bilateral, electrolytic lesions of the perirhinal cortex or the fornix, combined lesions of both these structures, or sham operations. Following recovery from surgery, rats were retested on the preoperatively learned discrimination problems, as well as learning a new discrimination and discrimination reversal. Results indicated that all animals with lesions exhibited temporally graded retrograde amnesia, whereby memories acquired in the recent past (1-3 weeks) were impaired, and memories acquired in the remote past (6-8 weeks) were spared. There was no difference in the magnitude of retrograde amnesia between the three lesion groups. Animals in the perirhinal, fornix, and combined lesion groups were able to learn a new discrimination problem at a rate comparable to control rats; however, the animals with lesions were impaired at learning the discrimination reversal. The perirhinal, fornix, and combined lesion animals also exhibited a significantly faster forgetting rate over a 2-week retention interval than control rats. These results suggest that medial temporal structures including the perirhinal cortex and the fornix are involved in the consolidation of mnemonic information and that their involvement in this process occurs over a discrete period of time. PMID- 10456102 TI - Late-training amnesic deficits in probabilistic category learning: a neurocomputational analysis. AB - Building upon earlier behavioral models of animal and human learning, we explore how a psychobiological model of animal conditioning can be applied to amnesic category learning. In particular, we show that the late-training deficit found in Knowlton, Squire, and Gluck's 1994 study of amnesic category learning can be understood as a natural consequence of Gluck and Myers's (1993) theory of hippocampal-region function, a theory that has heretofore been applied only to studies of animal learning. When applied to Knowlton et al.'s category learning task, Gluck and Myers's model assumes that the hippocampal region induces new stimulus representations over multiple training trials that reflect stimulus stimulus regularities in the training set. As such, the model expects an advantage for control subjects over hippocampal-damaged amnesic patients only later in training when control subjects have developed new hippocampal-dependent stimulus representations; in contrast, both groups are expected to show equivalent performance early in training. A potentially analogous early/late distinction is described for animal studies of stimulus generalization. Our analyses suggest that careful comparisons between early and late-training differences in learning may be an important factor in understanding amnesia and the neural bases of both animal and human learning. PMID- 10456103 TI - How big is human memory, or on being just useful enough. PMID- 10456104 TI - Nonassociative and associative modification of head-waving produced by aversive tentacular stimuli in Aplysia. AB - Head-waving, a spontaneously occurring exploratory and appetitive behavior of the marine mollusc Aplysia, provides an opportunity to examine mechanisms of learning expressed in a nonreflexive behavior. The present study explores nonassociative and associative forms of learned modification of head-waving produced using an aversive stimulus as reinforcement. Experiments on intact, freely behaving animals demonstrate that training with electric shock as an aversive unconditioned stimulus, delivered unilaterally to the anterior tentacles, produces a learned shift in head-waving behavior away from the side on which shock was applied. This behavioral change is a novel learned behavioral response that is influenced by the topographic location of an aversive stimulus. Furthermore, training with application of tentacle shock reinforcement, contingent upon the animal's head position, produces operant conditioning of head waving. Thus, anterior tentacle shock is effective as an aversive reinforcer for both nonassociative and operant learning expressed in the head-waving behavior of Aplysia. PMID- 10456105 TI - Multiple forms of facilitation produced by aversive tentacular stimuli in cerebral ganglion sensory neurons of Aplysia. AB - Aversive tentacular stimuli can produce both nonassociative and associative modification of head-waving behavior of Aplysia. Sensory neurons (the J/K cluster SNs) in the cerebral ganglion of Aplysia constitute an afferent pathway for aversive stimuli of the anterior tentacles. We used intracellular recording to examine plasticity in these neurons, particularly side-specific or site-specific alterations, which may be involved in mediating aspects of the learning induced by tentacle shock. The results of these experiments indicate that the J/K SNs exhibit several forms of plasticity: (1) Post-tetanic potentiation (PTP); (2) heterosynaptic facilitation; (3) activity-dependent synaptic facilitation; and (4) side-specific spike broadening. Furthermore, by activating SNs directly to produce PTP and producing heterosynaptic facilitation with tentacular stimuli that were either inside or outside the receptive field of individual SNs, it was possible to dissect and analyze the differential contribution of intrinsic SN activity and heterosynaptic modulation to activity-dependent facilitation induced by behaviorally relevant stimuli. Collectively, these data raise the possibility that plasticity in primary afferent SNs may be involved in US processing during learning induced by tentacle stimulation. PMID- 10456107 TI - Suppression of c-fos induction in rat brain impairs retention of a brightness discrimination reaction. AB - Recently, the induction of transcription factor-encoding immediate-early genes such as c-fos was observed in distinct brain regions of rats trained to acquire a footshock-motivated brightness discrimination in a Y-maze. The functional relevance of inducible transcription factors for learning and memory formation is, however, not clear. To address this question in the present study, we have used a synthetic antisense phosphorothioate oligodeoxynucleotide to suppress in vivo the expression of c-fos in rat brain. Intrahippocampal application of the oligodeoxynucleotide 10 hr and 2 hr before starting a brightness discrimination training drastically reduced the induction of c-Fos immunoreactivity normally observed in limbic and cortical areas after the training session. Acquisition of the discrimination reaction was not affected by this treatment. In a relearning test 24 hr after the first training, retention of the discrimination reaction was specifically impaired compared with rats pretreated with control oligodeoxynucleotide or saline. Our findings are consistent with the hypothesis that the inducible transcription factor c-Fos is involved in processes underlying the formation of long-term memory. PMID- 10456106 TI - An autonomous kinase generated during long-term facilitation in Aplysia is related to the Ca(2+)-independent protein kinase C Apl II. AB - Prolonged treatment with serotonin leads to long-term facilitation of sensory-to motor neuron synapses in Aplysia. We have shown previously that there is a protein synthesis-dependent increase in an autonomous kinase activity that phosphorylates a protein kinase C substrate during an intermediate phase of this facilitation. Here, I report that the increase in autonomous activity was independent of RNA synthesis, suggesting it may play a role in the maintenance phase of synaptic facilitation. Immunoprecipitation experiments using an antibody specific to the Ca(2+)-independent protein kinase C, Apl II, demonstrated that the autonomous kinase activity increased by serotonin emanated from Apl II. Chelerythrine, an inhibitor targeted to the substrate binding site of protein kinase C, also blocked the autonomous kinase activity increased by serotonin. Using immunoblotting experiments and calphostin-C, an inhibitor targeted to the regulatory domain of protein kinase C, the autonomous activity is shown not to be a catalytic fragment of Apl II. Furthermore, a higher concentration of calphostin C was required to inhibit autonomous kinase activity than regulated kinase activity, suggesting that calphostin-C's binding site in the regulatory domain of Apl II is modified in the autonomous kinase. These data suggest that an autonomous kinase derived from Apl II may play a role in synaptic facilitation in Aplysia. PMID- 10456108 TI - Modification of hippocampal synaptic proteins by nitric oxide-stimulated ADP ribosylation. AB - Nitric oxide has been shown to be an important neuronal signaling molecule that participates in both behavioral and synaptic plasticity. To better understand the potential mechanisms by which NO regulates synaptic function, the ability of NO to stimulate the modification of synaptic proteins by ADP ribosylation was examined. Two NO donors, sodium nitroprusside and 3-morpholinosydnonimine, stimulated the ADP ribosylation of proteins at apparent molecular masses of 42, 48, 51, 54, and 74 kD in hippocampal synaptosomes. This stimulation was likely owing to the production of NO by the donors; ADP ribosylation was not stimulated by non-NO decomposition products of sodium nitroprusside, and quenching of superoxide anion did not inhibit Sin-1-induced ADP ribosylation. Experiments using NAD+ that was radiolabeled on the nicotinamide moiety demonstrated that the modification of proteins of molecular masses of 30, 33, and 38 kD are not true ADP ribosylation, whereas labeling of the 42-, 48-, 51-, 54-, and 74-kD proteins likely represent ADP ribosylation. Some of the substrates were brain specific (54 and 74 kD), whereas others (42 and 51 kD) were present in multiple nonbrain tissues. PMID- 10456109 TI - The Drosophila mutation turnip has pleiotropic behavioral effects and does not specifically affect learning. AB - The Drosophila mutant turnip (tur) was isolated on the basis of its poor performance in an olfactory learning task, and also has a reduction in protein kinase C (PKC) activity. PKC has been found in the nervous systems of a wide range of organisms and appears to have an important role in learning and memory related processes. Unfortunately, previous reports documenting the learning defect of tur lacked the controls required to assess the origins of the poor performance of the mutant. We have analyzed the effects of the tur mutation on both associative and nonassociative learning as well as on PKC activity. Additionally, the effects of the mutation on the task-relevant sensorimotor abilities of the flies were assessed. Although we were able to replicate previous behavioral and biochemical results obtained with tur, we discovered that the tur mutation also affected response to electric shock and caused a drastic reduction in the locomotor ability of the flies. Because locomotion is an essential component of the learning assays, this result makes it impossible to conclude that tur specifically affects learning and demonstrates the crucial importance of sensorimotor controls in conditioning experiments. PMID- 10456110 TI - The cerebellum, LTD, and memory: alternative views. PMID- 10456111 TI - Preserved performance by cerebellar patients on tests of word generation, discrimination learning, and attention. AB - Recent theories suggest that the human cerebellum may contribute to the performance of cognitive tasks. We tested a group of adult patients with cerebellar damage attributable to stroke, tumor, or atrophy on four experiments involving verbal learning or attention shifting. In experiment 1, a verb generation task, participants produced semantically related verbs when presented with a list of nouns. With successive blocks of practice responding to the same set of stimuli, both groups, including a subset of cerebellar patients with unilateral right hemisphere lesions, improved their response times. In experiment 2, a verbal discrimination task, participants learned by trial and error to pick the target words from a set of word pairs. When age was taken into account, there were no performance differences between cerebellar patients and control subjects. In experiment 3, measures of spatial attention shifting were obtained under both exogenous and endogenous cueing conditions. Cerebellar patients and control subjects showed similar costs and benefits in both cueing conditions and at all SOAs. In experiment 4, intra- and interdimensional shifts of nonspatial attention were elicited by presenting word cues before the appearance of a target. Performance was substantially similar for cerebellar patients and control subjects. These results are presented as a cautionary note. The experiments failed to provide support for current hypotheses regarding the role of the cerebellum in verbal learning or attention. Alternative interpretations of previous results are discussed. PMID- 10456112 TI - A neural model of cerebellar learning for arm movement control: cortico-spino cerebellar dynamics. AB - A neural network model of opponent cerebellar learning for arm movement control is proposed. The model illustrates how a central pattern generator in cortex and basal ganglia, a neuromuscular force controller in spinal cord, and an adaptive cerebellum cooperate to reduce motor variability during multijoint arm movements using mono- and bi-articular muscles. Cerebellar learning modifies velocity commands to produce phasic antagonist bursts at interpositus nucleus cells whose feed-forward action overcomes inherent limitations of spinal feedback control of tracking. Excitation of alpha motoneuron pools, combined with inhibition of their Renshaw cells by the cerebellum, facilitate movement initiation and optimal execution. Transcerebellar pathways are opened by learning through long-term depression (LTD) of parallel fiber-Purkinje cell synapses in response to conjunctive stimulation of parallel fibers and climbing fiber discharges that signal muscle stretch errors. The cerebellar circuitry also learns to control opponent muscles pairs, allowing cocontraction and reciprocal inhibition of muscles. Learning is stable, exhibits load compensation properties, and generalizes better across movement speeds if motoneuron pools obey the size principle. The intermittency of climbing fiber discharges maintains stable learning. Long-term potentiation (LTP) in response to uncorrelated parallel fiber signals enables previously weakened synapses to recover. Loss of climbing fibers, in the presence of LTP, can erode normal opponent signal processing. Simulated lesions of the cerebellar network reproduce symptoms of cerebellar disease, including sluggish movement onsets, poor execution of multijoint plans, and abnormally prolonged endpoint oscillations. PMID- 10456114 TI - The effects of reversible inactivation of the red nucleus on learning-related and auditory-evoked unit activity in the pontine nuclei of classically conditioned rabbits. AB - The pontine nuclei carry auditory conditioned stimulus information to the cerebellum during classical conditioning of the nictitating membrane response in rabbits. In well-trained animals learning-related as well as stimulus-evoked unit activity can be recorded throughout the pontine nuclei but particularly in the lateral and dorsolateral pons. Recent work in our laboratory has provided evidence that the learning-related unit activity in the pons is dependent on the interpositus nucleus and that the pons is not a site of essential plasticity for the learned response. In the present study we considered the question of whether learning-related unit activity might be projected from the interpositus nucleus to the pons through the red nucleus, a primary output target of the interpositus and a structure known to be essential for expression of the learned response. Multiple unit recordings were taken from lateral and dorsolateral pontine locations in well-trained rabbits before and during cooling of the red nucleus. Analysis of pooled data for all recording locations within the lateral and dorsolateral pons indicated that reversible inactivation of red nucleus abolished both stimulus-evoked and learning-related unit activity. However, we also found discrete recording locations where stimulus-evoked and learning-related unit activity were attenuated but not abolished by red nucleus cooling. PMID- 10456115 TI - The learning-related activity that develops in the pontine nuclei during classical eye-blink conditioning is dependent on the interpositus nucleus. AB - A growing body of research now implicates the cerebellum in the formation and storage of the critical neural plasticity that subserves the classically conditioned eye-blink response. Previous anatomical, physiological, and behavioral research suggests that auditory-conditioned stimulus information is routed to the cerebellum by the pontine nuclei. However, it has also been observed from multiple unit recordings that some populations of pontine cells, in addition to showing auditory-evoked responses, also show changes in activity that is learning-related. It is unknown whether this learning-related activity is generated by the pontine cells or whether it is generated by some other structure and projected to the pontine nuclei. Because the cerebellum has been implicated in the formation of the essential plasticity that subserves this learned behavior, we examined how multiple unit recordings of learning-related activity within the pontine nuclei are affected by reversible inactivation of the interpositus nucleus of the cerebellum. The results indicated clearly that when the interpositus nucleus was inactivated, the learning-related activity in the pontine nuclei was abolished completely and the auditory stimulus-evoked activity was unaffected. In contract, when the facial nucleus was inactivated, both the auditory stimulus and the learning-related activity were still present. These results indicate that the learning-related activity exhibited by some populations of pontine nuclei cells is dependent on the interpositus nucleus and may represent feedback from the cerebellum. PMID- 10456116 TI - Reversible inactivation of the cerebellar interpositus nucleus completely prevents acquisition of the classically conditioned eye-blink response. AB - Numerous studies from several laboratories report that temporary inactivation of the cerebellar interpositus nucleus and regions of overlying cortex during eye blink conditioning completely prevents acquisition of the conditioned eye-blink response (CR) without affecting the ability to learn the CR in subsequent training without inactivation. Recently, these results have been challenged by the suggestion that learning was not completely blocked in these studies. Instead, it has been suggested that low levels of responses on test sessions might represent a retarded form of learning caused by drug effects on cerebellar cortex. The present study was designed to address this issue directly. Very low doses of muscimol were used to selectively inactivate the interpositus nucleus of rabbits during five conditioning sessions. Animals performed no significant levels of CRs during those sessions. Training was continued four more sessions without any inactivations to test whether any learning had occurred during the previous five sessions. Detailed analysis of responses during session six revealed that learning was completely blocked by the low doses of muscimol infused into the interpositus during the first five sessions. Animals subsequently acquired the CR normally. These results confirm and extend the original findings that appropriate lesions (either temporary or permanent) of the interpositus nucleus completely prevent acquisition of the conditioned eye-blink response. Other issues regarding reversible inactivation studies are also discussed. PMID- 10456117 TI - Acquisition of a new-latency conditioned nictitating membrane response--major, but not complete, dependence on the ipsilateral cerebellum. AB - Classical conditioning of the nictitating membrane response (NMR) of rabbits is simple associative learning of a motor response. In several two-stage experiments, reversible inactivations of the deep cerebellar nuclei in stage 1 appeared to prevent acquisition of NMR conditioning in naive rabbits--no conditioned responses (CRs) were evident after inactivations were lifted in stage 2. Results of a three-stage experiment were different. When subjects were first trained with a light conditional stimulus (CS) in stage 1, reversible cerebellar inactivations during conditioning to a different, tone CS during stage 2 did not appear to prevent new learning because CRs to the tone CS were evident when the inactivation was lifted. Results from the two-stage experiments support the suggestion that the cerebellum is essential for the acquisition of NMR conditioning, but results from the three-stage experiment do not. Here, we use a three-stage design with different interstimulus intervals (ISIs) in stages 1 and 2. Because CRs develop with latencies-to-peak dependent on the ISI, learning during stage 1 can be dissociated from that accruing in stage 2. Complete inactivation of the ipsilateral cerebellar nuclei with muscimol substantially but not completely prevented learning with the second ISI during stage 2 because small CR peaks around the stage 2 ISI could be detected in some subjects after the inactivation had been lifted in stage 3. We suggest that the weak levels of conditioning possible during unilateral inactivation depend on the contralateral cerebellum or on extracerebellar circuitry and that these may be capable of supporting transfer of conditioning in a previous three-stage experiment. But, we confirm that normal NMR conditioning is critically dependent on the ipsilateral cerebellum. PMID- 10456118 TI - Persistent phosphorylation parallels long-term desensitization of cerebellar purkinje cell AMPA-type glutamate receptors. AB - This study is aimed at testing the hypothesis that sustained phosphorylation underlies long-term desensitization of AMPA receptors, which is thought to be the mechanism of long-term synaptic depression in cerebellar Purkinje cells (PCs). We induced long-term desensitization of AMPA receptors in rat cerebellar slices by (1) a 4-min bath application of quisqualate (0.1 mM) or (2) a 15-min bath application of a protein kinase C (PKC) activator, phorbol-12,13-diacetate (0.5 microM) or -dibutyrate (0.6 microM), followed by a 4-min AMPA (0.1 mM) application. In slices so treated, labeling with an antibody (12P3) against a peptide corresponding to part of AMPA receptor subunit GluR2 including serine 696 and phosphorylated at this serine site revealed phosphorylation of the AMPA receptors in PC dendrites that was sustained for at least 1 hr. At an early phase, within 20 min after the chemical stimulation, the phosphorylation was resistant to an Ca2+ chelator (BAPTA-AM), a metabotropic glutamate receptor antagonist (MCPG), and a PKC inhibitor (calphostin C), whereas at a late phase, 30 min or more after the chemical stimulation, it was blocked by these reagents similarly to long-term desensitization of AMPA receptors. Taken together with data obtained previously using different protocols of chemical stimulation, the present results strongly support the above-mentioned hypothesis. PMID- 10456119 TI - Binding reaction of aflatoxin B1 with immunoglobulin G against aflatoxin B1 bovine serum albumin complex. AB - Polyclonal immunoglobulin G antibodies were raised against aflatoxin B1-bovine serum albumin complex and characterised. Antibodies against the complex were obtained after a single intradermal multiple site injection of water in oil emulsion of the complex into adult female albino rabbits. Equilibrium dialysis and Scatchard plot analysis of the interaction of the antibodies with aflatoxin B1 showed that the antibodies have high affinity for binding aflatoxin B1. The average number of binding sites on the antibody molecules for aflatoxin B1 is 1.74 +/- 0.20 with mean standard free energy change (delta F1(0) of -23.10 KJ/mol, while the average association constant is 2.35 +/- 0.73 x 10(-4)M-1. Male wistar strain albino rats after immunization with the complex showed lower mortality when challenged with a single dose of aflatoxin B1. The results suggest that such antibodies with high affinity for aflatoxin B1 could be used in the immunointerception of the toxin. PMID- 10456120 TI - Schistosoma haematobium infection in Abeokuta. AB - Ten years after the initial report of the outbreak of schistosomiasis in the vicinity of Sokori stream (a tributary of Ogun River) in Abeokuta, Nigeria, investigation was carried out to determine the present status of the disease, given the increased socioeconomic activities in the city. A total of 1670 school pupils in three schools within one kilometer radius of the stream, and 234 adults found using the stream for some water related activities were examined for eggs in their urine. One hundred and ninety-eight (11.9%) of the pupils and 98 (7.7%) of the adult group were infected. Compared to results of earlier surveys 1985 (55%) and 1987 (60.9%), there was a considerable reduction in the prevalence of infection. Also, 20.1% of pupils examined in the 1987 survey excreted between 200 and 1000 eggs/10 ml urine while the highest egg count in this survey was 189 eggs/10 ml urine. In addition to the improved water supply observed in the city, there has also been increased alternative recreational sites which may have reduced the propensity of the children to swim in the stream. Three snail species were seen (Bulinus forskalii, Physa waterlotti, and Melanoides voltae), none of which was naturally infected with cercariae. PMID- 10456121 TI - Haematological values in pregnancy in Ibadan, Nigeria. AB - Haematological values were studied in 495 apparently healthy pregnant Nigerian women receiving antenatal care in a maternity hospital in Ibadan, Nigeria. Means, standard deviations and ranges of haematrocrit, total and differential white cell counts, absolute platelet count, erythrocyte sedimentation rte, prothrombin time (PT) and partial thromboplastin time (PTTk) were determined. The mean haematrocrit was 0.311 (SD 0.033), mean total leucocyte count was 5.765 x 10(9)/L (SD 1.753 x 10(9)/L) and mean platelet count was 193 x 10(9)/L (SD 46 x 10(9)/L). The mean erythrocyte sedimentation rate was 18.6 (SD 7.4) mm/hour, mean prothrombin time was 12.3 (SD 2.5) seconds and mean partial thromboplastin time was 38.4 (SD 2.0) seconds. There were significant differences between the second and third trimesters of pregnancy in nay of the haematological values studied. The study has presented the range of variations in haematological values (including those that measure coagulation) in apparently healthy pregnant Nigerian women. It would appear that in the population studied, there were no significant differences in these haematological values between the second and third trimesters in otherwise healthy women receiving regular antenatal care. PMID- 10456122 TI - Penetrating eye injuries in children in Ibadan. AB - A retrospective study was carried out on 60 children under 12 years of age with penetrating eye injuries. This formed 28.7% of all penetrating eye injuries seen in the University College Hospital, Ibadan. Corneal lacerations extending to the limbus was the main type of injury occurring in 65% of the patients. In all, 63% of the patient were males. Visual outcomes was poor in this age group with only 19.3% (11 patients) retaining good vision of 6/12 or better. PMID- 10456123 TI - Some aspects of the pattern of systemic hypertension in the adult population of Gassim, Saudi Arabia: age distribution of the subsets of hypertensives. AB - As part of our studies in prevent nephrology, we have recorded causal blood pressure during a total population cross-sectional survey of the Faizia East Primary Health District of Buraidah (FEPHD), capital of Gassim region of Saudi Arabia. Out of 5671 subjects whose blood pressure could be recorded, 2222 were above 19 years of age and constituted our adult population. For this report, hypertension have been defined as equal to and greater than 140/90 mm Hg. Total hypertensive population had been divided into three subsets, based on this basic definition, namely combined systolic and diastlic hypertension (S/DHPN), isolated systolic hypertension (ISHPN) and isolated diastolic hypertension (IDHPN), a format which is not generally clear in previous studies on the subject. The overall prevalence of systemic hypertension was found to be 23.58%, which is lower than the figure of 36% for USA (pre-primary prevention intensive campaign). It is assumed that all the three subsets mentioned above have been used in calculating the latter. Although males tended to be more hypertensive than females (OR = 1.22; Chi Square = 3.89; P = 0.05; C.I.: 1.00 < OR < 1.49), the marginal difference (25.7% vs 22.10%) was largely due to the IDHPN subset (OR = 1.73; Chi Square = 4.48; P = 0.034; C.I.: 1.01 < OR < 2.96). In both the S/DHPN and ISHPN: statistical significance was not achieved. Significantly, ISHPN subset constituted the bulk--56.68%--of the hypertensive population while IDHPN constituted the least--11.64%. When analysed into 10-year-age cohorts, ISHPN showed the steepest rise in prevalence with age. Rather suprisingly, the IDHPN did not rise with advancing age while the S/DHPN slope was in between the two. We are unable to identify this pattern in our literature search on the subject. We cannot assess its significance by this study, but we wonder whether or not it carries any prognostic significance in terms of target organ damage. It is possible that this pattern may be peculiar to the sub-region but it certainly is not spurious and in our view deserves further scrutiny. We wish to suggest that clear definition of the three subsets of hypertensive population should be mandatory when defining prevalence. This may provide some further clues in the prognosis and pathogenesis of target organ damage. PMID- 10456124 TI - The origin(s) of the periarticular osteophytes of osteoarthritic knee joints. AB - Oestophites obtained from the margins of 25 osteoarthritic knee joints undergoing replacement have been investigated using a battery of tinctorial stains. The microarchitecture of the osteophyte appears to be identical to that of the normal articular cartilage. Other findings suggest that oesteophytes are derived from hyperplasia of the surface chondrocytes at the 'marginal transitional zone'. PMID- 10456125 TI - Studies of preventive nephrology: self-urinalysis as a feasible method for early detection of renal damage. AB - The ever expanding pool of ESRF patients is exerting considerable strain on the health care resources of all nations of the world. Rationing, in one form or the other has therefore become the norm for most countries. Because dialysis prolongs life and is more readily available, and because ethically acceptable donor kidneys remain in short supply, thus limiting the potential of renal transplantation, this rather exasperating situation is bound to continue unless the entry point into the pool can be actively tacked. As part of our initial effort in this direction, we have examined the feasibility of self urinalysis by the general population as an epidemiological tool for detecting evidence of early renal damage by a total population cross-sectional survey of Faizia East Primary Health District (FEPHD) of Buraidah, capital city of the Gassim region of Saudi Arabia. Out of a de facto population of 7,695, 75.37% (5,800) cooperated fully. Majority of those who could not cooperate (881 [11.44%]) were infants and children. A total of 969 subjects (12.59%), mostly males at work, were not available. Only 45 (0.58%) subjects refused to participate. Housewives were significantly more amenable to the organisation of family self-urinalysis than head of the family (92.2% vs 61.4%; chi square = 321.78; df: 3; P < 0.0001). The mean family size was 7.82 (+/- SD: 3.82). Above the age of 4 years, 66.5% of males (2108/3170) as against 81.7% of females (2,641/3232) were able to carry our self-urinalysis. 11.76% of boys and 8.5% of girls below the age of 5 years were able to carry out self-urinalysis. Only 0.05% of male subjects and 0.03% of females failed to interpret colour change for proteinuria correctly. Similar remarkable competence was demonstrated for glycosuria by the population. We conclude that self-urinalysis is quite feasible in the general population, even if illiterates, if young. It can form a sound foundation, if properly harnessed, for a renal registry. PMID- 10456126 TI - Ocular morbidity survey in an opthalmically underserved rural area of Nigeria. AB - This paper describes in some detail a prevalence of blindness study conducted in a rural area of South-Western Nigeria, which is served by a peripheral eye clinic. By a stratified random cluster-sampling procedure, 1975 people in ten villages were selected for ocular examination. Community prevalence rate of ocular morbidity was 3.3% with the highest prevalence in the 51-60 years age group. There was a male-female ratio of 4:3 and the prevalence of ocular morbidity in the 31-50 years age group was most significant, but people in this age group seldom reported at the peripheral eye clinic serving the survey areas. Age-related macular degeneration was found to be prevalent in the community, while few cases were reported at the peripheral eye centre. The information gathered in the survey is being used to plan and improve eye care services from the peripheral eye clinic. Case finding in persons aged 31-50 years and those at risk for age-related macular degeneration has been recommended as part of the programme of the peripheral eye clinic. PMID- 10456127 TI - Orofacial indices: a study in 240 Nigerian children. AB - This paper seeks to establish normal orofacial indices in Nigerian children under the age of 12. The children were grouped according to age and sex. The indices studied were: mouth width/upper vermilion are with lip relaxed; coefficient of upper lip curvature; upper lip height/mouth width; height of cutaneous upper lip/upper lip height; height of upper vermilion/upper lip height; upper lip elasticity; and size of oral aperture. The indices varied with age. Except for upper lip elasticity in 10-11-year-old children, there were no significant differences in orofacial indices in males and females. The coefficient of upper lip curvature which is an index that measure the curve or protrusion of the upper lip, was maximum in two or three-year-olds in both sexes. The orofacial indices calculated from lip dimensions could provide important baseline information for surgical repair or reconstruction of the orofacial region in black children. PMID- 10456128 TI - Greater sciatic notch in sex differentiation in Nigerian skeletal samples. AB - A total of 150 X-ray films (A-P view) of the pelvis of adult (90 male and 60 female) Nigerians were used to measure the width, depth, posterior segment, total and posterior angles of the greater sciatic notch. Index 1 was calculated by dividing the greatest depth OC by the greatest width AB and multiplying by 100. Index 2 was calculated by diving the posterior segment OB by the greatest width (AB) and multiplying by 100. These indices were used to study the sex differences in the hip bones of these Nigerians. Demarking points of these parameters and indices were worked out to determine sex. The width, depth, total angle and index 1 were insignificant in sex determination. Posterior angle and index 2 were found to be the most useful assigning sex to 74.5% of the male and 90% of the female bones. PMID- 10456129 TI - Correlation of smoking and coffee drinking with sperm progressive motility in infertile males. AB - The objective of the study was to assess the effect of some social characteristics on the forward progressive motion of sperms in males of infertile couples. The study was conducted in the Department of Obstetrics and Gynecology, King Kahlid University Hospital (KKUH), Riyadh, from September 1, 1994 to February 28, 1995. Semen specimens were obtained from 68 Saudi males of infertile couples, and 29 fertile males. Using computer-assisted semen analysis (CASA), sperm forward progressive motion was determined and graded on a scale of 1-4. The actuarial life table survival analysis method was used to investigate the sperm motility potentials across intervals 0-1, 1-2, 2-3, and 3-4 and the effect of smoking and coffee drinking was also determined on the sperm motility. Smoking was found to significantly improve sperm motility among the infertile group. Similarly, frequent coffee drinking was shown to enhance sperm motility. There is a need for further studies on the influence of smoking and coffee drinking, as well as the plethora of other influence of the daily life that may compound the detection of the subtle changes in semen quality. PMID- 10456130 TI - Infertility and other fertility related issues in the practice of traditional healers and Christian religious healers in south western Nigeria. AB - Traditional healers have been an established source of health care delivery in Africa for centuries while Christian religious healers (193 traditional healers and 99 Christian religious healers) with respect to infertility and some other fertility-related issues. The findings show that both types of healers believe that infertility is most commonly due to the past life of the woman, physical problems related to the womb or to male potency, and imcompatibity between the man and the woman. Traditional healers also believed that being bewitched or being cursed can lead to infertility. Both groups of healers threat infertility by sacrifices, prayer and fasting, and timing of intercourse to coincide with the fertile period. Also 61% of traditional healers and 87% of religious healers advice their-clients with infertility to do nothing at least initially. To those clients seeking advice on preventing pregnancy, traditional healers tend to recommend herbal concoctions, beads and rings while Christian healers tended to recommend condoms, withdrawal method and the safe period. Both groups are consulted on premarital sex, premarital conception, sex during pregnancy and influencing the sex of an unborn baby. It was concluded that both traditional healers and Christian faith healers are involved with infertility and other fertility-related issues in their practices. There is an overlap in beliefs about causes and treatment of such conditions among both groups although areas of differences in beliefs and practices are clearly identifiable. PMID- 10456131 TI - Maternal and neonatal thyroid status in Saki, Nigeria. AB - The influence of inadequate iodine intake on maternal and neonatal thyroid status in Saki, a locality known to have environmental iodine deficiency and located within the goitre belt of Southwestern Nigeria was investigated. One hundred and five (105) subjects and ninety (90) controls from Ibadan were recruited into the study. All subjects were apparently healthy with at least thirty-seven (37) completed weeks of gestation. Each subject was assessed for goitre and blood sample obtained by venepuncture for thyroid function tests before onset of labour. Spot urine samples were also obtained from the mothers at the time of examination for urinary iodine estimation. At delivery, cord blood samples were obtained from the babies for thyroid function tests. The mean maternal urinary iodine excretion was significantly less in Saki when compared with Ibadan (144.76 +/- 14.68 vs 213.36 +/- 9.89 ug/L P < 0.01). Maternal total goitre rates (TGR) were 46.8% and 2.2%, while maternal visible goitre rates (VGR) were 26.6% and 1.1% in Saki and Ibadan, respectively. Maternal mean plasma total T3 and total T4 were relatively lower in Saki as compared to Ibadan, though the differences were not statistically significant. Mean maternal plasma TSH in Saki was slightly above our laboratory reference interval (0.3-5.0 mU/L) and higher than the level seen in Ibadan mothers (6.54 +/- 1.58 vs 4.18 +/- 1.37 mU/L,), though the difference was also not statistically significant. Mean neonatal total T3 and T4 were lower in Saki when compared to Ibadan, but the differences were not statistically significant. The mean neonatal plasma TSH in Saki was above the adult reference interval and significantly higher than the level seen in Ibadan (9.82 +/- 1.64 vs 4.18 +/- 1.17 mU/L, P < 0.05). The incidence of neonatal chemical hypothyroidism (NCH) in Saki was 14.7 per 1000 babies. No case of NCH was seen in Ibadan. These results suggest environmental iodine deficiency and relative chemical hypothyroidism of mothers and neonates in Saki when compared to mothers and noenates from non-iodine deficient areas. PMID- 10456132 TI - Chloroquine reduces fertilizing capacity of epididyma sperm in rats. AB - Chloroquine diphosphate was administered intraperioneally to two groups of adult male rats at two dosage levels of 5 mg base/kg body weight and 10 mg base/kg body weight. The drug was administered over a period of fourteen days. Fertilizing ability of epididymal sperm was assessed in the rats by cohabitation with normal cycle female rats and foetal size was noted. Epididymal spermatozoa were counted in these groups of rats. Chloroquine effected a dose-related reduction in fertility of the male rats as evidenced by a reduction in average number of foetuses of cohabited females, although the sperm count in the groups was not significantly different. This suggest that the drug does play an inhibitory role in sperm function. PMID- 10456133 TI - Abnormal serum alkaline and acid phosphatase isoenzymes in female breast cancer patients. AB - Serum total, different isoforms of both alkaline and acid phosphatases, liver function enzymes, calcium, inorganic phosphate, heamatocrit, white blood cells and platelet counts were determined in 50 female patients suffering from breast cancer. The serum total alkaline and total acid phosphatases within the breast cancer group were variable with significant elevation of both enzymes compared with the corresponding control values. The activities of alanine and aspartate transferases were higher than the control values, while the decreases in serum albumin and heamatocrit were statistically significant. In the breast cancer patients, the increases in the activities of both heat and urea labile alkaline phosphatases were significant. Similarly, in the patients, the tartrate-labile acid phosphatases activity was significantly elevated while the difference in tartrate resistant activity was not significant. In 9 patients (18%), both total alkaline and acid phosphatases were excessively raised when compared with the control. The increased activities of urea-labile and heat-labile alkaline phosphatases as well as tartrate-resistant acid phosphatases are suggestive of increased activities of osteoclast and osteoblasts associated with bone metastasis. A possible diagnostic importance of this observation deserves further investigation, using monoclonal antibody techniques. PMID- 10456134 TI - Coagulation profile in healthy Nigerian neonates. AB - A study of some coagulation factors were carried out in preterm and term infants on the first day of life. Screening coagulation tests--prothrombin time (PT), partial thromboplastin time with kaolin (PTTK) and the concentrations of Factors VIII: C and fibrinogen were determined in 100 normal newborn infants classified into three groups according to their gestational ages: 28 to 30 weeks, 31 to 36 weeks, 37 to 42 weeks. The respective values were compared with those of six month-old infants as well as adults. The mean values of the screening coagulation tests, PT and PTTK, Factor VIII: C and fibrinogen were significantly different in all the three gestational age groups of nonates when compared with those of six month-old infants and adult Nigerians. This suggests that a relative hypocagulable state exists among newborn infants and could be responsible for increased bleeding tendencies in these groups of infants. This is the first documented report of coagulation profile in Nigerian neonates, and the range obtained in this study can thus be regarded as standard for healthy Nigerian neonates. The body of data should therefore provide a basis for evaluating newborn noenates with bleeding problems. PMID- 10456135 TI - Evaluation of lacidipine (a calcium blocker) in the treatment of hypertension in black African people: a double-blind comparison with hydrochlorothiazide. AB - Diuretics and calcium blockers are particularly effective in the treatment of hypertension in Blacks, who, characteristically, have low-renin hypertension. The efficacy and tolerable of lacidipine (a calcium) blocker of the dihydropyridine class) and hydrochlorothiazide (a diuretic) were compared in a 12 week double blind randomised parallel group study of Nigerians with essential hypertension. Lacidipine was given at a starting dose of 4 mg daily by mouth and increased to 6 mg if there was no satisfactory response at 4 weeks, and hydrochlorothiazide was stared at 25 mg daily by mouth and increased to 50 mg if necessary. Twenty-four patients (8 male) in the lacidipine group and 17 (5 male) in the hydrochlorothiazide group were evaluable at the end of the trial. In the lacidipine group, SBP was significantly reduced from 157 +/- 14 mmHg to 146 +/- 24 mmHg (P < 0.00001) and DBP from 90 +/- 9 mmHG to 87 +/- 15 mmHg (P < 0.00001) with BP normalisation rates of 67% and 79% at 4 weeks and 12 weeks, respectively. In the hydrochlorothiazide group, SBP was significantly reduced from 16.4 +/- 19 mmHg to 141 +/- 17 mmHg (P < 0.00001) and DBP from 102 +/- 6 mmHG to 89 +/- 7 mmHg (P < 0.00001) with normalisation rates of 77% and 82% at 4 weeks and 12 weeks respectively. The groups did not differ in BP reduction nor normalisation rates. There were no reported side effects. PMID- 10456136 TI - Endemic goitre prevalence in Ifedapo Local Government Area of Oyo State, Nigeria. AB - As part of efforts aimed at providing an adequate data base for the control of iodine deficiency disorders (IDD) in Nigeria, we undertook an extensive goitre survey of Ifedapo Local Government Area (LGA) of Oyo State, which lies within the goitre-belt of South-western Nigeria. Primary school pupils aged between six and twelve years were the subjects of the survey. From each of ten health districts of the LGA, one primary school was included in the survey. For each school, the entire school population was assessed for goitre by palpation and goitre graded according to WHO/ICCIDD criteria. The survey included 3599 pupils (1889 males and 1710 females). The total goitre rate (TGR) for the LGA was 23.4% while the visible goitre rate (VGR) was 6.4%. The highest goitre rate was recorded in District I (Sango, Saki) TGR = 39.5%; VGR = 17.3%), while the lowest rate was seen in District 4 (Wasengare) TGR = 5.2%; VGR = 0.7%). Females had higher goitre rates than males (TGR: 25.7% vs 21.3%; VGR: 7.5% vs 5.5%). Most of the goitres were Grades IA (36.7%) or IB (35.7%). Goitre grades 2 and 3 accounted for only 27.3% and 0.2%, respectively. These results show that there is a mild to moderate IDD problem in Ifedapo LGA. The implications of this finding, in view of the known deleterious consequences of IDD, are discussed and the need for correction highlighted. PMID- 10456137 TI - Oxygen injector ventilation in adults and children: use of a variable pressure driving gas valve and a right angled connector. AB - An oxygen injector system (Harwill Medical, South Africa) was tested for ventilating intubated patients. There were 21 adults and 25 children. The system incorporates a variable pressure reducing value and a right-angled end connection with a 14 gauge oxygen exit nozzle and a 12 mm side port to atmosphere. Intravenous propofol was used to maintain anaesthesia, while the patients underwent alternate 10-minute periods of ventilation with the oxygen injector or an Ohmedia 7000 ventilator. Tidal volumes (ml/kg) achieved by mechanical ventilation were 8.06 +/- 1.89 in adults and 7.28 +/- 2.01 in children, compared with 5.79 +/- 2.13 and 4.57 +/- 1.58 respectively, with the oxygen injector (P > 0.05). Peak airway pressures (cmH2O) on mechanical ventilation were 18.61 +/- 3.06 and 20.21 +/- 2.89 in adults and children. The corresponding pressures with the oxygen injector were 19.42 +/- 3.97 and 18.49 +/- 3.27 (P > 0.05). Cardiovascular parameters were unchanged. A side port adaptation provided a suction capability for water at a rate of 9.16 ml/s. We conclude that this system is an effective universal ventilator suitable for short-term ventilation and suction of patients. PMID- 10456138 TI - Cutaneous malignant melanoma: Tabuk experience. AB - Cutaneous melanoma (CM) has a rising morbidity and mortality in the western world but is rare in certain geographical areas including the Middle East. The aim of this study is to define the pattern of CM in this environment over a period of about two decades. A review of associated histological reports, dermatology, plastic general surgical admissions and outpatient census statistic of the North West Military Hospital (N.W.A.F.H.) were carried out from January 1978 to June 1996. The clinico-therapeutic information from both the review case and newly discovered CM was then studied. The result shows that CM is probably rare in the Tabuk military environment and possibly has a low mortality among the affected individuals. The presence of only 2 cases of CM among 73,955 patients over about 20 years suggests that this neoplasm is rare in N.W.A.F.H. Surgery, with localised expert reconstruction, probably offers the best cure for uncomplicated CM in this area. It is suggested that the geographical environment, genetic attributes, custom, attitude, presence of white, painted, sun-reflecting buildings, traditional dress-code and behaviour of the indigenes probably contribute to the suppression of and protection against CM in Tabuk. It is recommended that regular, antimlanoma education awareness programmes among the indigenes and avoidance of sunbathing attitude of the expatriate community should be encouraged in order to maintain this suggested natural selection protection. PMID- 10456139 TI - Effects of glucagon, glucose, adrenaline and insulin infusion on blood glucose level in the common African toad (bufo regularis). AB - Male toads, weighing 70-123 g, were divided into 13 groups with 8 toad in each group. Animals in each group were fasted overnight before the experiments. Toads in groups 1 to 4 were infused for 30 minutes with adrenaline, 5 ugkg-1 min-1, glucose, 5.5 mg kg-1 min-1; glucagon 2 ugkg-1 min-1; and insulin 2000 uU kg-1 min 1, respectively. Blood samples for blood glucose measurement were taken before, during and after each infusion. The experiment was repeated in groups 5 and 6 using 3.5 mg kg-1 min-1 and 7.5 mg kg-1 min-1 of glucose respectively. Toads in groups 7 to 9 were pretreated with prazosin, 0.2 mg/kg and those in groups 10 to 12 were pretreated with propranolol, 0.5 mg/kg. After pretreatment, glucose, glucagon and insulin infusions were repeated in the alpha-blocked and beta blocked toads, respectively. Group 13 was infused with 0.7% saline and served as the control. The results showed that infusions of adrenaline, glucose and glucagon resulted in significant hyperglycaemia while insulin caused hypoglycaemia. The hyperglycaemic response to glucose was dose-dependent. The experiments using blockers showed that the glycaemic effects of glucagon and insulin are mediated via beta adrenoceptors, that for glucose is via alpha adrenoceptors and from an earlier study, the glycaemic response of adrenaline is through both alpha and beta adrenoceptors. PMID- 10456140 TI - The influence of iodine deficiency on the cognitive performance of school children in Saki, south-west Nigeria. AB - The influence of inadequate iodine intake on the thyroid status and cognitive performance of school children in Saki, a town within the goitre belt of South western Nigeria with known environmental iodine deficiency, was assessed. One hundred and ninety-seven (197) study subjects from Saki and seventy (70) appropriately matched control subjects from Moniya, near Ibadan, the Oyo State capital were recruited into the study. The subjects were apparently healthy primary school pupils between the ages of 10 and 14 years and had been resident in the respective locality for at least two (2) years. After establishing the presence of goitre, blood was collected for thyroid function tests by venepuncture from each pupil. The cognitive function tests, Draw-A-Person (DAP) test and the Standard Progressive Matrices (SPM) were administered simultaneously to all the pupils in the class. Urine samples were collected for urinary iodine estimation. The mean urinary iodine concentration was significantly lower in Saki than in Moniya (134.81 +/- 69.86 vs 220.00 +/- 69.00 ug/L, P < 0.01). The total goitre rates (TGR) and the visible goitre rates (VGR) were 15.2% and 1.5% for Saki, and 8.6% and 4.3% for Moniya, respectively. The mean plasma total T4 was significantly lower in Saki pupils (97.55 +/- 26.64 nmol/L vs 122.52 +/- 26.51 nmol/L P < 0.05). The TSH level was higher in Saki pupils than in Moniya pupils (4.72 +/- 1.38 mU/L vs 4.26 +/- 1.28 mU/L), but the difference was not statistically significant. Scores on intelligence function tests (DAP and SPM) were lower for Saki pupils when compared with Moniya pupils, though the differences were not statistically significant. These results show that there is a mild but significant iodine deficiency disorder problem in Saki. However, the children in Saki still maintained euthyroidism and the mild degree of iodine deficiency did not seem to have adversely affected their cognitive performance. PMID- 10456141 TI - Sterological estimation of seminiferous tubular dysfunction in chloroquine treated rats. AB - We have used the simple-point sampling of linear intercept lengths and the optical dissector method to describe the effects of chloroquine on rat testicular morphology. The rats were injected intraperitoneally with chloroquine phosphate (CQ), an antimalaria and amphiphilic drug known to induce generalized lipidosis. The result showed that CQ (10 mg/kg/day) treatment for 7 weeks significantly reduced (i) the weight of the testis, (ii) the daily sperm production count, (iii) the total spermatoid count per testis and (iv) the star volume of the seminiferous tubules. There was however an increase in the total spermatocyte count per testis. Together, these findings suggest that disruption of spermatogenesis accompanied a decrease in tubular size in CQ treated rats. PMID- 10456142 TI - Blood pressure pattern in Saudi population of Gassim. AB - Blood pressure pattern for the Kingdom of Saudi Arabia has not been defined. In a total cross-sectional population survey of the Faizia East Primary Health District of Buraidah, capital of Gassim region, Saudi Arabia, 5671 subjects out of a de facto population of 7695 got their blood pressure recorded. The study district consisted of an urban section and a rural sector. Mean systolic and diastolic blood pressure were computed for the total population in conventional 5 year age cohorts as well as in an arbitrary functional age groups. The curve pattern and trends of the percentiles, were defined. Mean blood pressure (systolic and diastolic) was found to rise with age in both genders (male SBP: r = 0.66, P < 0.000001: female SBP: r = 0.58, P < 0.00001; male DBP: r = 0.53, P < 0.00001; female DBP: r = 0.45, P < 0.00001) and to correlate significantly with BMI. Both systolic and diastolic values were consistently higher in females than males. Overall means (+/- SD) were: for SBP, male 109.9 (+/- 21.57) vs female 114.33 (+/- 21.22) mm Hg; df: 5669: P < 0.00000; CI: -5.5, -2.0; for DBP, male 62.85 (+/- 16.89) vs female 64.67 (+/- 14.99) mm Hg; df: 5669; P < 0.0000; CI: 2.65, -0.989. Proteinuria (macroalbuminuria) was found to correlate positively and very significantly with both systolic and diastolic blood pressure: for SBP: r = 0.074, P < 0.0001; DBP: r = 0.055, P < 0.0001. Perhaps more significantly, in the context of preventive nephrology, is the observation that the intercept of the regression line with blood pressure was below the level conventionally regarded as hypertensive, suggesting that nephron damage may have occurred at this lower level. Overall mean diastolic blood pressure but not systolic was found to be significantly higher in the rural environment than the urban setting: mean rural DBP: 66.43 (+/- 15.699) vs urban: 62.78 (+/- 78); P < 0.00001; rural SBP: 113.71 (+/- 23.95) vs urban: 112.69 (+/- 19.87), P = NS. No discernible effect of consanguinity in marriage on blood pressure could be detected in this study but we believe that further details are required before a definite statement can be made on this important subject. The percentiles presented can only be regarded as foundation figures requiring further validation before they can be useful in determining cut-off levels for hypertension for the Saudi population. PMID- 10456143 TI - Body mass index (BMI) in the Saudi population of Gassim. AB - In a total cross-sectional population survey of the Faizia East Primary Health District of Buraidah, Gassim region of Saudi Arabia, 6,044 (2727 male and 3317 females) subjects out of a de facto population of 7695 got their BMI computed because infants and restless or bedridden subjects could not be examined. Mean (+/- SD) and percentiles (25th & 75th) were calculated in the conventional 5-year age cohorts as well as in functional age groups, namely, 0-5, 6-12, 13-49, 50-69 and 70+ years. 5th, 10th, 25th, 50th, 75th, 90th and 95th percentiles were computed only for the functional age groups. In general, the trend was for BMI to increase with age in both genders but the curve pattern showed some plateauing from about the age of 50 with slight decline in later life. Females had significantly higher indices than males, this becoming quite prominent from the 10-14 year age cohort. This difference persisted irrespective of the types of age grouping or residential location. Overall means (+/- SD) were 20.14 +/- 5.98 vs 22.22 +/- 7.21 for males and females respectively; df: 5771; p = 0.0000; 95% CI: 2.43, -1.735. Subjects in the urban living environment had significant higher indices than their rural counterpart: (21.666.92 vs 20.446.33: df: 5771; P = 0.0000; 95% CI: 1.595, -0.840). From the age of 15 about one quarter of females are overweight (BMI at the 75th percentile > 25) and from 30 years the same proportion are frankly obese (BMI > 30). Both systolic and diastolic blood pressure were significantly positively correlated with BMI in both genders: male SBP: r = 0.22, P < 0.0001; male DBP: r = 0.21, P < 0.00001; female DBP: r = 0.18, P < 0.00001. PMID- 10456144 TI - Atypical beta-adrenoceptors mediate isoprenaline induced vasodilatation in the rabbit uterine vascular bed. AB - beta-Adrenoceptor subtype(s) mediating isoprenaline-induced vasodilator responses in the perfused rabbit uterine vascular bed was characterized. Isoprenaline induced dose-dependent reduction in perfusion pressure with -logED50 value of 5.8 +/- 0.1. Propranolol (10(-6) M) had little or no effect on isoprenaline-induced vasodilatation. Similarly, cyanopindolol (10(-6) M) did not antagonize the vasodilator response. CGP 12177, an agonist on beta 3- or beta 2- adrenoceptor mediated responses. The results would indicate that isoproterenol-induced vasodilation in the perfused rabbit (oestrus or pregnant) uterine vascular bed was mediated by atypical beta-adrenoceptors which may not be identical with those in adipose tissues or gastro-intestinal tract. In addition, the fact that pregnancy did not enhance isoprenaline-induced vasodilation would suggest that these receptors may not be involved in the hyperaemia of pregnancy. PMID- 10456145 TI - Replantation of avulsed teeth: considerations and complications. AB - Avulsion of teeth is a traumatic dental injury which can be managed by replantation. Important considerations for successful replantations are highlighted. Several sequelae to replantation are discussed in relation to the clinical features observed in a patient seen 6 years after replantation of two avulsed upper incisors. Many of these complications might have been avoided if the patient had returned to the clinic for root canal therapy shortly after the teeth was replanted. PMID- 10456146 TI - An unusual cause of bladder rupture in a paraplegic. AB - Over distention of the bladder or a bladder substitute may lead to rupture. The chances of rapture are more in a previously scarred bladder, particularly in a patient with poor sense of bladder fullness such as paraplegic. PMID- 10456147 TI - Squamous cell carcinoma of the cervix simulating an advanced malignancy of the ovaries. AB - A 40-year-old woman was diagnosed as having stage II squamous cell cervical carcinoma and managed with radiotherapy. Three months after treatment, she presented with features suggestive of an advanced ovarian tumour including gross abdominal swelling, bilateral ovarian tumours, multiple tumour seedlings in the abdominal cavity and ascites. There was also pleural effusion. Operative findings revealed widespread intra-abdominal metastases whose histology, contrary to expectations, showed squamous cell carcinoma of cervical origin. Distant metastases from squamous cell carcinoma of the cervix are rare. A high index of suspicion is necessary to detect this unusual mode of presentation. PMID- 10456148 TI - Auto-amputation of the penis in HIV infected patient. PMID- 10456149 TI - Muscarinic blockade with oral hyoscine palliates angiotensin converting enzyme inhibitor induced cough. PMID- 10456150 TI - Relationship of the ECG with ventilatory function tests in chronic obstructive lung disease (COLD) in Nigerians. AB - Although the most objective assessment of the severity of COLD is done by spirometer, this is not as readily available in Nigeria as the electrocardiograph, thus the ECG is often used to assess it. This study examines the relationship between the ECG findings and FEV 1 obtained by spirometry and expressed as a proportion predicted for age and sex in 92 Nigerians with COLD of which 39 had associated pulmonate. There was an inverse relationship between the mean electrical axis of the P wave on the ECG and the FEV 1 of predicted. The axis was more positive (82 +/- 12 degrees) among those with FEV 1 less than 50% of predicted (group 1) and this decreased progressively with increasing FEV 1 to 72 +/- 14 degrees in those with FEV 1 between 50-70% (group 2) and 67 +/- 16 degrees in those with FEV 1 greater than 70% (group 3). There was also a significant relationship between FEV 1 and the development of cor pulmonale and 71.4% of those who had the lowest FEV 1 (group 1) had cor pulmonale while the proportion decreased with the reduction in the severity of airway obstruction. A rightward P wave axis greater than 80% was statistically significant associated with development of cor pulmonale (X2 = 29.5 (P < 0.001) of those with COLD and cor pulmonale, 72.2% had a P wave axis greater than 80%. Only 13.2% of those with COLD alone had a P wave axis greater than 80 degrees. Cardiac arrhythmias were present in 28 patients, ventricular (VA) in 23 cases, and atrial (AA) in 7 cases. Both AA and VA were associated with more severe airway obstruction. All 3 patients who had atrial fibrillation (AF) and FEV 1 below 50% of predicted (group 1) and had cor pulmonale while 2 of the remaining 4 with AA also were in group 1 and the remaining 2 in group 2. The distribution of ventricular arrhythmias (VA) showed that 10 (43.5%) were in group 1, 9 (39.1%) in group 2, and 2 (8.7%) in group 3. Both VA and AA were also significantly related to the development of cor pulmonale. A P wave amplitude greater than 2.5 mm was significantly related to the development of clinical cor pulmonale X2 = 24.6 P < 0.001. The results show that ECG findings correlate well with spirometric assessments of FEV 1 and clinical severity in Nigerians with COLD and are, therefore, useful where spirometry is unavailable. PMID- 10456151 TI - Experience of cognitive behaviour therapy in psychiatric practice in Nigeria: 1: The model and the method. AB - The paper highlights the need for knowledge of the principles and methods of cognitive behaviour therapy (CBT) among Nigerian therapists, the historical development of the model, the theoretical framework of the model, and the behavioural and cognitive techniques in common use. Although the method is limited in its use to educated people, the number of such persons is rising in our population. CBT meets the African patients need to be directed and the therapists need for brevity. PMID- 10456152 TI - Plasmid profiles of multiple drug resistant local strains of Staphylococcus aureus. AB - In an attempt to determine the molecular basis for the degree of resistance among strains of S. aureus to Beta-lactam antibiotics, 50 clinical isolates of coagulase-positive strains of S. aureus were obtained. The strains and a control strain, Oxford NCTC 6571, were screened for the presence of Beta-lactamase, R plasmid, and sensitivity to six Beta-lactam antibiotics. The molecular weight of the isolated R-plasmid DNA was determined by agarose gel electrophoresis. Forty five of the 50 strains of S. aureus exhibited single or multiple drug-resistance. Plasmid DNA isolated from 13 of the resistant strains of S. aureus had the same molecular weight of 21 kb or 13.63 MDa, while one of the 13 strains and 3 others had R-plasmids with a higher molecular weight. PMID- 10456153 TI - A new prognostic scoring system in neonatal tetanus. AB - A prospective study of 75 cases of neonatal tetanus was carried out with a view to developing a prognostic scoring system which can be used to assess the severity of the disease and to serve as a basis for comparing results of different therapeutic interventions from various centres. The case fatality rate was 77.3%. A table was designed containing 6 parameters and a rating of 1 to 4 was given to each parameter in decreasing order of severity. The minimum total score attainable was 6 and the maximum 24 (the severity of the disease was inversely proportional to the score). The differences between the means of each of the parameters among patients who died and the survivors were significant (P < 0.005). Regression analysis showed that each of the parameters independently had a significant effect on the total score (P < 0.005). Using the percentage mortality at each score, a pattern emerged such that total scores of 6-11 indicated severe tetanus (mortality rate 100%); 12-17, moderate disease (mortality rate 68%), and total scores of > 17 indicated mild neonatal tetanus (mortality rate 18%). Associated poor prognostic factors identified included pneumonia, recurrent apnoea, cyanosis, and opisthotonus. PMID- 10456154 TI - The planetary positions and relationships at the dates of birth of a cohort of Nigerian schizophrenics. AB - Some astrological hypotheses related to predisposition to severe mental illness were tested by analysing the zodiacal signs, the interactions between planetary qualities (aspects), and the occurrence of full and new moon dates, on the dates of birth of 221 schizophrenics, compared with 112 normal subjects. The sun signs of the schizophrenics were significantly more likely to be in the signs associated with introversion, while those of the control population were significantly more likely to be in the outgoing signs. A significantly higher proportion of schizophrenics had their Mars (i.e., symbol of aggressiveness) in the outgoing signs than the normal population. A significantly higher proportion of control subjects fulfilled operational criteria for adequacy of number of aspects between the sun and the other planets. The tendency for a higher proportion of schizophrenics to have "difficult" aspects just failed to reach significance. A significantly higher proportion of control subjects had aspects between the sun and mars; and also a significantly higher proportion of control subjects had "soft" (helpful) aspects between the sun and mars. These findings are in keeping with the well-known oddity of schizophrenia (schiz = split; phren = mind); such that, a group which collectively is characterised by an "introverted" self (i.e. sun sign), has a coexisting aggressive tendency (i.e. strong mars) and poor integration between the elements of the psyche and the self (i.e. inadequacy of aspects between Sun and other planets). However, the findings give only partial support to key astrological postulates because there was a non significant trend for more schizophrenics to be born in "water" signs and on full moon dates. PMID- 10456155 TI - Blastocyst implantation: effect of thyroidectomy and thyroxine replacement in the rat. AB - The long term effects of thyroidectomy and chronic thyroxine treatment on blastocyst implantations in female albino rats were studies. Implantation was delayed in both thyroidectomized and hyperthyroid rats, as there was no evidence of blastocyst implantation on day 6 of pregnancy. By day 9, when signs of implantation were observed, the number and weights of the implantation sites were significantly reduced in the athyroid and hyperthyroid rats. Both thyroidectomy and chronic thyroxine treatment significantly reduced the fetal sites and increased the incidence of resorptions when compared with the control. It is concluded that a functional thyroid gland is required for normal implantation to occur. PMID- 10456156 TI - Pattern of bacterial pathogens in surgical wound infections. AB - Wound swabs from surgical patients were studied from 1989 to 1991 to review the pattern of nosocomial infection in the University College Hospital, Ibadan, Nigeria. The prevalence rate of nosocomial infection was 4.9%. The ratio of gram negative to gram-positive organisms in wound infection was 3:1 with klebsiella species and Pseudomonas species emerging as the most important gram-negative organisms. Staphylococcus aureus was the single most prevalent organisms in surgical would infections. Recommendations on control measures are given. PMID- 10456157 TI - Gall bladder disease and ABO blood group. AB - Because it had been shown that an elevated total serum cholesterol was associated with phenotype A in many western European and Japanese populations as compared to non-A phenotype, a study was conducted to see whether there was any correlation between ABO blood group and gall bladder disease in our environment. No correlation was found between ABO blood group and gall bladder disease. However, the incidence of gall bladder disease was much higher in females than in males and the incidence was found to be highest in the fourth and fifth decades of life. PMID- 10456158 TI - Contrasting influence of the living environment and gender on systemic hypertension in Saudi population of Gassim, Saudi Arabia. AB - In a cross-sectional total population survey of Faizia East Primary Health District of Buraidah, which is divided into an "urban" and "rural" sections, casual blood pressure was recorded in 5671 subjects. 2222 (910 males) were adults (> 19 years). 3299 (1561 males) were between 3-18 years, making the paediatric/adolescent cohort of the population. The rest were below 3 years and were not included for calculations in this report. For adults, HPN was defined as > or = 140/90 mm Hg and P/A it was > or = 95th percentile for the age cohorts 3 5, 6-9, 10-12, 13-15 and 16-18 years as recommended by The Task Force for Blood Pressure Control in Children (1987). Each of the three subsets of HPN were derived from these basic definitions without modification. In the adults population, overall prevalence (S/DHPN + ISHPN + IDHPN) was 23.58% (524/2222) and gender prevalence was marginal in favour of males (25.71 vs 22.1% for males and females respectively; OR: 1.22, chi square = 3.89; p = 0.05; CI: 1.00 < OR < 1.49). By contrast, overall HPN prevalence in P/A was 10.64% (351/3299) and in all the age cohorts, girls were very significantly more hypertensive than boys. Overall figures were 13.06 vs 7.94% respectively for girls and boys; OR: 0.57; chi square = 22.65; p < 0000019; CI: 0.45 < OR < 0.73. Only in severe HPN (> or = 99th percentile) in 16-18 year age cohort did male preponderance become obvious. When the influence of the living environment was examined, in adult population (urban: 40,001; rural: 1670 subjects), for all age cohorts the rural environment which is largely inhabited by the unsophisticated Bedouins living close to their livestock, significantly predisposed to HPN as compared to the urban setting. Overall figures are 19.95 vs 32.60%, urban vs rural respectively: OR: 0.52; chi square = 40.4; p < 0.000000; CI: 0.42 < OR < 0.64. On the other hand, in the P/A population (2301 urban, 998 rural) for the age cohorts 3-5 and 6-9 years the urban environment significantly predisposed to HPN. At 10-12 years the prevalence was virtually even (12.65 vs 12.71%; OR: 0.99; p: NS). Thereafter, i.e. 13-15, 16 18 year cohorts, the rural environment became more dominant, just like the adult pattern. In other words whatever the factors in the unsophisticated Bedouin "rural" setting which predisposes to HPN does not become operative until after childhood--after 12 years. This contrasting gender and environmental influence in our study population seems unique, being reported for the first time. It may be pointing us in new direction in the elucidation of the pathogenesis of HPN and should deserve further studies. PMID- 10456159 TI - Determinants of single limb stance balance performance. AB - This study was designed to evaluate the effects of gender, physical activity level, age and anthopometric indices on the single limb stance balance performance of asymptomatic subjects. Eighty physically active and 120 sedentary subjects (100 males and 100 females, age ranged, 12-40 years) from Ile-Ife community were recruited for the study. The maximum duration that each subject was able to maintain balance whiled standing on their dominant leg with eyes open and eyes closed were monitored. It was found that: 1. Males had a higher (P < 0.01) balance time than females; 2. The physically active subjects performed better (P < 0.001) on the eyes closed test than sedentrary subjects of comparable age, weight and height; and 3. The balance time, with eyes open or closed, improved with chronological age and increased with height and body weight. The findings in this cross sectional study suggest that exercise programmes could be used to improve balance performance. It was concluded that the single limb stance time testing protocol described in this study could be used: a. As a screening tool in population based studies to identified individuals with a balance dysfunction; and b. In clinical practice to monitor improvement in neuromuscular function of patients with injuries to the lower extremities undergoing rehabilitation. PMID- 10456160 TI - Detergent solubilization of liver microsomal acyl-coenzyme A:1-acyl glycerophosphocholine acyltransferase in nephrotic rat. AB - Hyperlipidaemia is a common feature of nephrotic syndrome and this has been thought to involve increased assembly and secretion of very low density lipoprotein (VLDL) in the liver. An important pathway for an indirect modulation of VLDL. Synthesis is the reaction catalyzed by the acyl-coenzyme A:1-acyl glycero-phosphcholine acyl transferase. We therefore investigated the activity of this enzyme in liver microsomes isolated from puromycin amino nucleoside induced nephrotic rats. When oleoyl-CoA was employed as the acyl-donor, our results indicated that both the total and detergent soluble enzyme activities (112.2 +/- 16.7; 116.1 +/- 17.5 units, respectively) were significantly higher than the corresponding control levels of 91.1 +/- 11.1 and 75.4 +/- 20.9 units respectively. The percentage stimulation by sodium cholate were 176.5 and 192.2 for the control and nephrotic rats, respectively. In absence of sodium cholate, when oleoyl CoA was replaced by arachidonoyl-CoA as acyl-donor, the measured total enzyme activity was only significantly reduced in the control rats (71.1 +/ 8.9 Vs 91.1 +/- 11.1 Units). Oleoyl-CoA as acyl-donor gave higher values for the soluble and residual enzyme activities (90.4 13.3; 99.5 34.5 unit) than the corresponding control levels (75.9 +/- 10.0; 50.5 +/- 34.0 units) as compared to arachidonoyl-CoA. In the control group the difference was only significant in the residual activity (92.9 20.5 Vs 64.7 24.1 units). The addition of monomethylethanomine (200 mM) had little or no effect, while both reduced glutathione (10 mM) and 1,2-diacylglycerol (1 mM) caused significant reduction in measured activity. These results indicated that in nephrotic rats new phospholipid synthesis is enhanced and this could contribute to the increased VLDL assembly and secretion usually associated with nephrotic syndrome. PMID- 10456161 TI - Comparison of body temperatures taken at different sites and the reliability of axillary temperature in screening for fever. AB - This prospective study was carried out to compare temperatures taken at different sites and also to determine the reliability of axillary temperatures in screening for fever in healthy and febrile children under 10 years of age. The difference between the mean rectal and axillary temperatures was not significant in both healthy and febrile neonates (P > 0.05). However, in healthy and febrile children beyond the neonatal period the mean rectal temperature was significantly higher than the mean axillary temperature (P < 0.001), the difference between the mean axillary and oral temperatures was significant (P < 0.001) but there was no significant difference between the mean oral and mean rectal temperatures (P < 0.05). The mean axillary temperature stabilisation time was 3.8 minutes while the mean rectal and oral temperature stabilisation times were 1.6 minutes and 1.8 minutes respectively. The sensitivity of axillary temperatures in detecting fever was 98% among neonates but only 47% among children beyond the neonatal period. The specificity and positive predictive value were 100% each among both the neonates and children beyond the neonatal period. The negative predictive value was 98.7% among neonates while it was 64.4% amount the older children. It is concluded that unlike in older children axillary temperature in neonates correlates well with the core temperature and it is sensitive enough to detect fever. Axillary temperatures rather than rectal temperatures should be taken in neonates, while rectal or oral temperatures be taken in older children. When the axillary route is used, the thermometer should be left in place for at least 4 minutes. PMID- 10456162 TI - Malaria parasitaemia and immunoglobulin levels in paired maternal-cord sera from south western Nigeria. AB - Blood samples were collected from one hundred and sixteen parturient women and one hundred and seventeen umbilical cords at delivery for the detection of malaria parasitaemia and determination of total serum immunoglobulins (IgG, IgM and IgA). Immunoglobulin levels were measured by the single radial immunodiffusion method and the enzyme-linked immunosorbent assay for cord blood IgM. Malaria parasites were found in 2.6% (3/117) of cord blood and 22.4% (26/116) of maternal samples. Primiparae had the highest incidence and density of parasites compared with multiparae. A negative correlation was obtained between parasite density and parity of the parturient women (r = -0.54, P < 0.005). Mean cord blood IgG (P < 0.001) and IgM (P < 0.0001) were significantly lower than the mean maternal IgG and IgM. Maternal IgG (r = 0.65, P < 0.001) but not IgM (r = 0.09, P < 0.50) correlated with those of cord blood. Mean IgM (P < 0.001) but not IgG (P > 0.50) and IgA (P < 0.40) was significantly higher in malaria positive parturient women compared with malaria negative women. These data confirms the transplacental transfer of IgG across the placenta and the higher incidence of malaria parasitaemia in primiparae. The presence of IgM in cord blood samples suggest intrauterine sensitization of the foetus to common infections. PMID- 10456163 TI - Anthropometric indices in singleton term neonates and their possible value in predicting some metabolic problems: a preliminary study. AB - We studied possible sources of variations in selected anthropometric indices and their relationship to neonatal metabolic problems in seventy-four singleton neonates born at an approximate gestation of 40 weeks, by selected anthropometry. There were significant differences particularly in values obtained for birth weight, mid upper arm circumference, chest circumference and the Ponderal Index. At the moment it is not possible to set discriminant values on any of the anthropometric indices as this will require a much larger sample size, however the authors believe that the use of anthropometry in predicting metabolic problems in neonates has some potential and serial measurements of particularly chest circumference, mid upper arm circumference and the Ponderal Index could form the basis of future studies. PMID- 10456164 TI - Effect of acute acetyl-l-carnitine treatment on daytime melatonin synthesis in the rat. AB - The effects of acute treatment of young male rats with 350 mg/kg of acetyl-l carnitine (ALCAR) at 15.00 hr on the synthesis and secretion of melatonin by the pineal gland and the retinas were studied 1 hr after injection. The pineal N acetyltransferase (NAT) activity was unchanged when compared with the control. However, there was a non-significant but slight increase in the melantonin content of the pineal glands in the ALCAR-treated rats. The serum level of melantonim in the ALCAR-treated group was significantly higher (P < 0.05) when compared with the control. The melatonin content of the retinas in the ALCAR treated rats was significantly higher than the control (P < 0.03). This result suggests that the high melatonin content in blood 1 hr after treatment with ALCAR was not as a result of the increased synthesis and secretion rate of melatonin by the pineal gland but rather from extra-pineal source, retina. PMID- 10456165 TI - Antenatal complications in adolescent mothers aged below 14 years. AB - Antenatal complications in 340 booked and 710 unbooked adolescent mothers aged 12 14 years were evaluated over a 2-year period at the Specialist Hospital, Yola, Adamawa State, Nigeria. Emesis gravidarum was observed in 290 (85.3%) and 612 (86.2%) booked and unbooked mothers, respectively. While 112 (32.9%) booked mothers had malaria, this ailment was observed in 508 (71.5%) of unbooked mothers. Forty-six (13.5%) booked mothers suffered from anaemia as against 483 (68.0%) unbooked patients. It was observed that preeclampsia manifested in 62 (18.2%) booked and 158 (22.2%) unbooked mothers, while eclampsia occurred in 18 (5.3%) and 66 (9.3%) booked and unbooked mothers, respectively. The rates of premature deliveries were 16.20% in booked mothers and 22.82% in the unbooked group. Other notable complications observed in both groups include premature rupture of fetal membranes (PROM), preterm contractions, antepartum haemorrhage, and urinary tract infections. There were slightly higher frequencies of the above complications in 12-year-olds, and these decreased slightly towards the age of 14 years. Nine of the 10 above observed complications occurred more in the lower socioeconomic classes [3-4] than in the upper social economic classes [1-2] in significant proportions. PMID- 10456166 TI - Soft tissue rheumatism among Nigerians. AB - Sixty-eight Nigerians were seen, investigated and treated for soft tissue rheumatism. Most of the subjects had capsulitis of the shoulder, commonly associated with Diabetes mellitus and Hypertension. Other diagnostic groups were however seen. The treatment modalities were basically intra articular steroids, analgesics and physiotherapy. PMID- 10456167 TI - Efficacy of pefloxacin in acute gonococcal urethritis. AB - A clinical trial of pefloxacin as a single oral regimen in the treatment of acute uncomplicated gonococcal infection was evaluated in sixty-four male patients with clinical and laboratory diagnosis of gonococcal urethritis using a single dose of 800 mg pefloxacin. Of the 64 men, 59 (92.2%) were infected by (penicillinase producing Neisseria gonorrhoeae (PPNG) strains, while 5 (7.8%) had non-PPNG strains. The cumulative percentage cure for all infections was 84.4%. Pefloxacin is effective as a single dose therapy for acute gonococcal urethritis in males infected by penicillinase producing and non-penicillinase producing Neisseria gonorrhea in our area of study. PMID- 10456168 TI - Intra-oral squamous cell carcinoma in Ibadan: a review of 90 cases. AB - Squamous cell carcinoma (SCC) of the oral cavity is considered the commonest of all oral malignancies. Between the period January 1960 and December 1985, 385 cases of oral malignancies were histologically diagnosed at Ibadan; 102 out of these were SCC. There are very few up to date reports in the literature on the incidence of intra-oral carcinoma amongst the Nigerian population, particularly on intra-oral SCC. Therefore, this study analyzed the lesion's incidence according to sex, age, and site prevalence, and also compared the results with studies from other countries. PMID- 10456169 TI - Predictors of academic performance in the pre-clinical sciences: effects of age, sex and mode of admission at the Maiduguri Medical School. AB - The effect of age on admission, sex and mode of admission as correlates of academic performance in the final pre-clinical examination was investigated. Records of 222 medical students covering five academic years (1987-1992) were obtained and matched for each student with the result of the final pre-clinical examinations in Anatomy, Physiology and Biochemistry. Age significantly affected students' performance as those below 20 years and those above 24 years performed better than those with ages between 20 and 24 years. A higher proportion of the male medical students were successful in all the three subjects when compared with females. Medical students admitted through the Joint Matriculation Examination performed better than those admitted through the remedial science and direct entry programmes. The implications of these findings are discussed. PMID- 10456170 TI - Invasive and in situ carcinoma of the cervix in young Nigerians: a clinico pathologic study of 27 cases. AB - A study of young Nigerians aged 30 years and below with invasive and in-situ cervical carcinoma confirms the notion that cervical cancer occurs in all age group. All patients except one were married while all were parous. Forty four percent of the patients presented with stages III and IV disease and hence, were unsuitable for surgery. The youngest patient was aged 17 years, indicating the need tare cancer screening among adolescents and a vigorous family planning programme that emphasizes barrier contraception. PMID- 10456171 TI - Darier-White disease in a Saudi patient associated with systemic involvement. AB - A 22 year-old Saudi patient is reported who had Darier-White disease (Keratosis follicularis), presenting with the classical cutaneous manifestation of this disease in addition to systemic symptoms, including chronic renal failure, mental retardation, epilepsy, cataract and corneal opacities. PMID- 10456172 TI - Experience of cognitive behaviour therapy in psychiatric practice in Nigeria: II: Illustrative case histories. AB - In order to give practical guidance to interested therapists, the author uses four illustrative cases to show how the technique of cognitive behaviour therapy can be applied in the clinical setting in Nigeria. First, a detailed cognitive assessment of the predisposing, precipitating and perpetuating factors associated with the presenting complaints is carried out. This helps the patient to realise that the symptoms are predictable, and therefore potentially surmountable. Second, the patient and therapist jointly formulate the problem in a few itemised statements. Third, the patient and therapist jointly articulate a set of cognitions (statements of insight) by which the problem in each formulated statement can be overcome. Fourth, as homework exercises, the patient affirms the set of cognitions at least twice daily, as if instructing him/herself (self instructional training). This helps to make the insights received in therapy sessions to become deeply ingrained in consciousness, and positively affect behaviour in ordinary circumstances of daily living. Finally, a "diary of daily transformation" is articulated, with which the patient monitors the self, in order to see that insights obtained are applied in daily living experience. Educated, articulate and committed patients with circumscribed psychological problems can achieve significant improvement in 3-20 weekly sessions. PMID- 10456173 TI - Ovarian cancer associated with ovulation induction: a case report. AB - A case report of a 38 year old lady who developed ovarian malignancy following 3 cycles of ovulation induction therapy is presented. She was observed to have clinically normal ovaries at laparotomy for tubal infertility 12 months previously. Although direct causal link between ovarian stimulation and cancer has not been established yet, a case is made for increased monitoring of patients receiving ovulation induction medication by physicians. PMID- 10456174 TI - Problem-solving for better health: literature as a resource. PMID- 10456175 TI - Development and organization of the hypophysiotropic hypothalamus driving the pituitary-gonadal axis in the rhesus monkey. AB - The purpose of the present review is to describe, with particular emphasis on the rhesus monkey, the ontogeny and functional organisation of the hypothalamic GnRH pulse generator. Control of pituitary-gonadal axis in higher primates is provided by a group of some 1,000 GnRH neurons that are diffusely distributed throughout the hypothalamus. After synthesis of a prehormone and formation of the mature decapeptide, GnRH is released in the hypophysial portal circulation and stimulates FSH and LH production. PMID- 10456176 TI - Dynamics of steroid regulation of GnRH secretion during the oestrus cycle of the ewe. AB - The oestrus cycle of the ewe is characterised by a long luteal phase followed by a short follicular phase and these periods are related to the production by the ovary of two major steroids: progesterone and oestrogen. Progesterone exerts a strong inhibitory effect on GnRH secretion during the luteal phase by a mechanism which is still unknown. Using an oestrogen-free ovine model and the portal blood collection technique we have obtained new insights into this mechanism. While progesterone removal induces a rapid increase in GnRH pulse frequency, progesterone reinsertion inhibits GnRH release even faster: less than 50 minutes. This action of progesterone is specific to the gonadotrophic axis and is mediated through an action on the nuclear receptor. Interestingly, this rapid mechanism is also strongly dependent of prior exposure to both progesterone and oestradiol. During the follicular phase, the rise in circulating oestradiol induces a robust preovulatory GnRH surge. In the ewe, this positive feedback effect is mainly exerted by an action of oestradiol on the mediobasal hypothalamus. Finally, we have also obtained evidence that progesterone priming is important for the full expression of the positive feedback action of oestradiol on GnRH secretion. In summary, progesterone and oestradiol sequentially exert opposite feedback effects on GnRH secretion during the oestrus cycle of the ewe but there is also clear evidence that the systems affected by these steroids are intimately linked. PMID- 10456177 TI - Mechanism of GnRH receptor signaling: from the membrane to the nucleus. AB - The purpose of this review is to update the information concerning the intracellular effect of GnRH. Binding of GnRH to a G-protein coupled receptor leads to stimulation of Gq and/or G11 protein and to activation of phospholipase C beta. Inositol 1-4-5-triphosphate and early diacylylycerol are the second messengers required for conventional protein kinase C activation. Activation of phospholipase A2 and phospholipase D are also involved, as demonstrated by the liberation of Arachidonic Acid and Phosphatidic Acid. Pituitary cells also express atypical protein kinase C isoforms which mode of activation is not known. Hypothesis concerning transcriptional regulation are presented. PMID- 10456178 TI - Gonadotropin receptors. AB - Gonadotropin receptors belong to a subgroup of G-protein coupled receptors characterized by a large extracellular domain responsible for the binding of the hormone. Soluble, hormone-binding, alternative splicing variants of the LH receptor, are present in high concentration. A mannose rich precursor form of LH and FSH receptor is accumulated inside target cells. FSH receptors are addressed to the basolateral domain of cells through specific signaling mechanisms. Gonadotropin receptors are also present in endothelial cells of target organ vessels and are involved in hormone transcytosis. Various genetic abnormalities of these receptors (and of the GnRH receptor) are discussed. PMID- 10456179 TI - Physiologic and genetic insights into the pathophysiology and management of hypogonadotropic hypogonadism. AB - Initiation and maintenance of reproductive capacity is contingent upon the pulsatile secretion of gonadotropin-releasing hormone (GnRH) from the hypothalamus. While GnRH deficiency manifests itself clinically as hypogonadism, the time of onset of the defect in GnRH secretion (i.e. before or after the onset of puberty) determines the clinical presentation that is further modified by the severity of the defect. GnRH deficiency can be associated with structural lesions that interfere with the normal pattern of GnRH secretion through compression or destruction of critical areas of the median eminence of the hypothalamus. Idiopathic hypogonadotropic hypogonadism (IHH) has been described and is characterized by isolated GnRH deficiency in the absence of a hypothalamic or pituitary lesion. Typically, patients with hypogonadotropic hypogonadism and anosmia have been given the diagnosis of Kallmann's syndrome and those with normal olfaction, IHH. However, both entities can be present in the same family, demonstrating variability in phenotypic expression and raising questions about the usefulness of this distinction. GnRH deficiency may be inherited via autosomal dominant, autosomal recessive and X-linked modes of inheritance. Specific defects in a number of genes have now been found in association with GnRH deficiency in the human and including the KAL gene, the DAX gene and the GnRH receptor gene, but not the gene encoding GnRH itself. While these mutations account for only a small fraction of cases, genetic studies not only provide promise for future treatment, but also insights into the complexity of the GnRH neuronal system. The treatment of GnRH deficiency is highly successful whether designed for steroid hormone replacement or for fertility. The challenge in the treatment of these abnormalities is in the early and accurate diagnosis and the use of replacement therapies that are as close to physiologic as possible. PMID- 10456180 TI - Role of FSH in male gonadal function. AB - The production of male gametes depends on the concerted action of the two gonadotropins FSH and LH on the testis. The action of LH is mediated through the production of testosterone by the Leydig cells. Since male germ cells possess neither FSH nor androgen receptors, the action of FSH and testosterone occurs through the Sertoli cells. Although the precise function of these two hormones remains elusive, the existing evidence suggest that both FSH and testosterone are able to stimulate all phases of spermatogenesis. In the male FSH is required for the determination of Sertoli cell number, and for induction and maintenance of normal sperm production. The crucial role of FSH in male gonadal function has been clearly illustrated by the discovery of a patient with an activating mutation of the FSH receptor. This patient had been hypophysectomized because of a pituitary tumor and, under testosterone substitution was unexpectedly fertile in spite of undetectable serum gonadotropin levels and had fathered three children. In this patient we could demonstrate a heterozygous activating mutation of the FSH receptor which resulted in cAMP production independent of FSH stimulation. This finding represents the first description of an activating mutation of the FSH receptor and demonstrates that FSH alone maintains spermatogenesis in man. On the other hand, the effects of the lack of FSH action are unclear. Among five men with a homozygous inactivating mutation of the FSH receptor only one was infertile and spermatogenesis was variably affected in the others. However, serum inhibin B values in these men were not completely suppressed and serum FSH levels were only moderately elevated, indicating the possibility that FSH receptor function was not completely abolished by the mutation. Elimination of FSH action is a prerequisite to suppress completely spermatogenesis for contraceptive purposes, while administration of both LH and FSH is necessary to induce sperm production in patients with hypogonadotropic hypogonadism. Experimental immunization of male monkeys against FSH markedly reduced germ cell proliferation and even induced infertility. At the cellular level, FSH stimulates the cAMP-dependent activation of protein kinase A in Sertoli cells, but the molecular mechanism of FSH action is poorly understood. In the primate, the gonadotropin withdrawal achieved by administration of a GnRH antagonist leads to a premeiotic arrest of germ cell proliferation, probably due to inhibition of the mitotic division of A-pale spermatogonia. Therefore, FSH might be the prime inducer of spermatogonial proliferation, while the successive maturation process could proceed independently of FSH. In summary, clinical and experimental evidence support the concept of an irreplaceble role of FSH in the primate. Only the combination of FSH and testosterone, however, supports a qualitatively and quantitatively fully normal spermatogenesis. PMID- 10456181 TI - Effects of progesterone on human spermatozoa: clinical implications. AB - Progesterone is a physiological stimulus of human sperm acrosome reaction. The effects of the steroid, which is present in high levels in the cumulus matrix that surrounds the oocyte, are mediated by an increase of intracellular calcium concentrations, tyrosine phosphorylation of proteins, efflux of chloride and stimulation of activity of phospholipases. These effects are due to activation of a nongenomic pathway. Two different types of receptors for progesterone, distinct from the genomic ones, have been identified on the surface of human spermatozoa. We demonstrated that sperm responsiveness to progesterone is impaired in subfertile patients and that is strictly correlated to the ability of fertilize the oocyte. In addition, the determination of sperm responsiveness is predictive of fertilizing ability with a positive predictive value of 90% and can be clinically useful for the preliminary assessment of the male partner to select the appropriate assisted reproductive technique. PMID- 10456182 TI - Intragonadal regulation of male and female reproduction. AB - FSH and LH are vital endocrine regulators of gonadal growth and function. However, locally produced--paracrine--steroidal and nonsteroidal substances mediate and modulate gonadotrophin action on gonadal cells. The testis and ovary share obvious morphological and functional homologies. Optimal functioning of the spermatogenic (Sertoli cells) and follicular (granulosa cells) epithelia depends on both endocrine stimulation by FSH and paracrine stimulation by androgen. In each gland, inhibins and related factors produced by FSH-stimulated Sertoli (testis) cells or granulosa (ovary) cells reciprocally modulate LH-stimulated androgen synthesis in neighbouring Leydig (testis) or thecal (ovary) cells. Thus androgens and inhibins are "unisex" paracrine regulators that operate in similar ways in both ovary and testis. PMID- 10456183 TI - [Genes and ovarian insufficiency]. AB - Premature ovarian failure is defined as hypoestrogenism with elevated gonadotropins, occurring before the age of 40. The prevalence of this syndrome is 3% of the women population. In most cases its etiology remains unknown. In theory, ovarian failure may occur by the following mechanisms: the pool of primordial follicule is too low; the atresia ou apoptosis is increased in the ovarian follicle; the follicle maturation is interrupted. In the past few years, several genes have been implicated in the pathogenesis of human or murine premature ovarian failure. Those genes can be classified according to the potential mechanisms previously described. Atm and c-kit gene code for proteins involved in maintaining the pool of primordial follicles. Genes located on the X chromosome, genes involved in blepharophimosis, in galactosemia code for proteins involved in follicular atresia. The gonadotropins, their receptors, GDF9 and connexin 37 are involved in follicle maturation. A better understanding of the family of genes involved in oocyte and follicular atresia should allow a better understanding of premature ovarian failure. Up to now, the better care for those patients is to give them hormonal replacement therapy and suggest oocyte donation when they desire to be pregnant. PMID- 10456184 TI - [Physiopathology of polycystic ovary syndrome]. AB - The Polycystic Ovary Syndrome (PCOS) includes three phenotypic compartments, not always fully associated, consisting in hyperandrogenism, anovulation and metabolic syndrome, secondary to insulin resistance. The pathophysiological grounds lie upon two main components, i.e.: the theca-interstitial cell (TIC) and the granulosa cell (GC) dysregulations, the former accounting for hyperandrogenism and the latter for anovulation, and both of them being under the influence of hyperinsulinism. The former mainly results from an enzymatic overactivity, yielding an exaggerated output of androgens by the TIC, but the type(s) of enzymes as well as the genetic or adaptative nature of its (their) dysregulation are still controversial. The main consequence of the CG dysregulation is the follicular arrest just before the time of dominance. This might result from an intrinsic abnormality in CG, involving the IGFs and/or the Inhibin/Activin/Follistatin systems. Alternatively, the CTI might have deleterious effects on GC, mainly via the intra-ovarian hyperandrogenism. The latter should not be regarded any more as an atretogenic phenomenon. It is closely related to the two main morphological features of PCOS, i.e.: the stromal hyperplasia and the excessive follicular number. PMID- 10456185 TI - Genetic abnormalities in polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a very common endocrinopathy and is the major cause of anovulatory infertility. It is also associated with an increased risk of non insulin dependent diabetes (NIDDM) in later life. Despite the importance of PCOS to women's health, little is known about its aetiology. Because of the well known familial clustering of cases of PCOS, recent studies in our department have focused on clinical and molecular genetic studies in an attempt to identify key genes which may be involved in its aetiology. We have found evidence that a polymorphism in the regulatory region of CYP11a (encoding P450 cholesterol side chain cleavage, also an important enzyme in the steroidogenic pathway) is associated with and linked to PCOS. In examination of the insulin gene (INS), we have shown, in three separate populations, that class III alleles in the INS-VNTR (the minisatellite in the regulatory region of the insulin gene) are associated with PCOS. Variation in this element has also been implicated in the aetiology of NIDDM. We propose that PCOS is an oligogenic disorder in which a small number of key genes interact with environmental factors (notably dietary), the balance of which factors determine, the typically heterogeneous, clinical and biochemical phenotype. PMID- 10456186 TI - Embryonic implantation. AB - Embryo implantation is a progressive process which requires a communication between two different organisms, and consists of three consecutive phases: apposition, adhesion and invasion. It must be realized throughout the period called "window of implantation", which is characterized by morphological and biochemical changes in the endometrium, as the plasma membrane transformation and the presence of some specific adhesion molecules, chemokines, cytokines, growth factors, and invasive proteinases. All of then acting in a paracrine/autocrine manner, and drive by endocrine hormones. PMID- 10456188 TI - Reproductive phenotypes in the estrogen receptor-alpha knockout mouse. AB - The generation and initial characterizations of the ER alpha KO and has proved exciting, however, it is worth noting that this model is relatively new to the field. These mice will undoubtedly prove invaluable to future studies of steroid hormones in normal development and function. The ER alpha KO mouse has been and continues to be utilized to further study the role of estrogen action in the cardiovascular system. bone physiology, behavior, the immune system, neurophysiology, and adipogenesis. Furthermore, studies of the documented "non genomic" effects of estradiol and progesterone in the brain as well as ligand independent actions involving cross-talk with other signaling systems will be advanced with further investigations using the ER alpha KO mouse. And finally, the ER alpha KO in combination with the recently developed ER beta KO mouse will prove invaluable in distinguishing the different roles that may exist between the two receptors in the estrogen signaling system. PMID- 10456187 TI - Progress in ovarian stimulation. AB - It is uncertain whether patients who do not ovulate but fail to conceive following CC therapy behave differently during gonadotropin therapy. However, if the infertile patient with PCOS is resistant to anti-estrogen therapy, gonatropin treatment administered through low dose protocols should be considered. As compared to conventional regimens, complication rates are reduced despite similar efficacy. It should be recognized that patients with PCOS constitute a very heterogeneous group. Therapy outcome may benefit from improved classification. The identification of clinical characteristics that identify those women with anovulation who are unlikely to respond to clomiphene would permit earlier use of gonadotropin therapy and potentially offer major health and economic benefits. Furthermore, if ovarian responsiveness to ovulation-induction therapy could be predicted for an individual patient, it might be possible to devise regimens that reduced the risk of ovarian hyperstimulation and multiple pregnancy. PMID- 10456189 TI - [Cardiovascular diseases: from epidemiology to biology]. PMID- 10456190 TI - [Pathologic examination of the heart]. PMID- 10456191 TI - [Degenerative heart valve diseases]. PMID- 10456192 TI - [Pathology of heart valve prostheses]. PMID- 10456193 TI - [Pathology of vascular prostheses]. PMID- 10456194 TI - [Cardiovascular sudden death]. PMID- 10456195 TI - [Pathology of primary cardiac neoplasms]. PMID- 10456196 TI - [Large vessel vasculitides]. PMID- 10456197 TI - [Positive diagnosis and etiology of myocarditis]. PMID- 10456198 TI - [Diagnosis and major etiologies of infectious endocarditis occurring on native valves]. PMID- 10456199 TI - [Superficial vascular malformations: classification and histopathology]. PMID- 10456200 TI - [Apoptosis in the cardiovascular system]. PMID- 10456201 TI - Does blood lactate measurement have a role in the management of the critically ill patient? PMID- 10456202 TI - Renal tubular acidosis. PMID- 10456203 TI - Screening for aetiology of thrombophilia: a high prevalence of protein S abnormality. AB - We systematically screened for the aetiology of thrombophilia in 115 patients with venous, arterial and small vessel thromboses. Forty-one patients (36% of those we examined) suffering from a variety of thromboses, including deep vein thrombosis, pulmonary embolism, arterial occlusion, cerebral infarction, Moyamoya disease and ulcerative colitis, were characterized either with positive lupus anticoagulants or with decreased activities of protein S, protein C, antithrombin III and/or plasminogen. Eight mutation sites were confirmed in 11 thrombotic patients using gene analysis. Decreased protein S activity was found with a high incidence (23 out of 115) in Japanese patients who suffered from not only venous thrombosis but also arterial and small vessel thrombosis. We emphasize here the important role of protein S in the pathogenesis of thrombosis in the Japanese population. PMID- 10456204 TI - Monitoring of cardiac troponin I in patients with acute heart failure. AB - Cardiac troponin I (cTnI) and creatine kinase MB isoenzyme (CK-MB) were measured in the plasma of 37 patients with acute heart failure. Elevated plasma cTnI concentrations were found in 89% of acute heart failure patients (P < 0.001 compared with a normal population), while plasma CK-MB showed no significant difference (P = 0.09). During follow-up, serial measurements of cTnI and CK-MB were performed. In acute heart failure patients, improvement of the clinical profile was associated with declining cTnI concentrations, while deterioration of heart function was closely related to increasing cTnI. Plasma CK-MB activities remained within the normal range throughout the observation period. This preliminary study provides evidence of cardiac damage to functionally overloaded myocytes. cTnI may be a sensitive marker both for early detection of myocyte damage and for monitoring of function in patients with acute heart failure. PMID- 10456205 TI - Multisite study of a second generation whole blood rapid assay for cardiac troponin T. AB - We evaluated a second generation, qualitative, whole blood rapid assay for cardiac troponin T (cTnT), which employs a more cardio-specific troponin T mouse monoclonal capture antibody. Using quantitative cTnT enzyme-linked immunosorbent assay (ELISA) results as the benchmark for accuracy, we compared the performance of the second generation and the original whole blood rapid assays in 445 samples from patients with the following diagnoses, determined by medical record review: myocardial infarction, coronary bypass surgery, ischaemic heart disease, musculoskeletal disease, renal failure or other noncardiac conditions. Overall, concordance between the second generation cTnT Rapid Assay and the quantitative cTnT ELISA, compared using the McNemar test and a cut-off concentration of 0.1 microgram/L, was in the range 76-94% for each patient group. Using a receiver operating characteristic plot, the cut-off for the second generation cTnT Rapid Assay was in the range 0.06-0.08 microgram/L. We conclude that the second generation cTnT whole blood assay has a 2.5-fold lower analytical cut-off than the original rapid assay (0.2 microgram/L) and may represent a more sensitive clinical tool for the rapid triage and risk stratification of cardiac patients. PMID- 10456207 TI - Differences in rates of glycation (glycation index) may significantly affect individual HbA1c results in type 1 diabetes. AB - Ten type 1 diabetic patients recorded their daily home blood glucose values, pre- and post-prandially, for 12 weeks. Blood was collected weekly for HbA1c and total haemoglobin measurement. A rolling 28-day mean of all blood glucose values and a glycation index (the ratio of the HbA1c to the rolling mean blood glucose) were calculated. In the pooled patients' data, there was a large scatter of results about the HbA1c versus mean blood glucose regression line. There was less variation in the results of individual patients. The glycation indices showed marked inter-individual variation, and in 60% of patients there was an inverse relationship between glycation index and mean blood glucose, suggesting a non linear relationship between mean blood glucose and HbA1c. Patients should be monitored on the basis of their own previous results, and in some patients blood HbA1c may be a less sensitive index of mean blood glucose concentration at higher glucose levels. PMID- 10456206 TI - Diurnal variations in peripheral insulin resistance and plasma non-esterified fatty acid concentrations: a possible link? AB - Glucose tolerance becomes impaired towards the evening. Increased peripheral insulin resistance may be responsible, at least in part, for this effect. The mechanism for the diurnal variation in insulin sensitivity is undefined. It is, however, possible that variations in non-esterified fatty acids (NEFA) could contribute to this variation because NEFA have been implicated in the pathogenesis of insulin resistance. Therefore, we have investigated insulin sensitivity and plasma NEFA responses to insulin at 0830 h and 2030 h in nine healthy men by measuring arterialized plasma glucose and venous plasma NEFA concentrations during a short insulin tolerance test. The studies were standardized for a period of fasting, pre-test meal and exercise. Insulin sensitivity measured KITT was greater (P < 0.05) in the morning [(20 +/- 7) x 10( 3) mmol/L/min] than in the evening [(11.6 +/- 2) x 10(-3) mmol/L/min]. Fasting NEFA levels were lower (P < 0.01) in the morning (373 +/- 84 mumol/L) than in the evening (913 +/- 122 mumol/L). Following insulin, NEFA fell more slowly (P < 0.01) in the morning (149 +/- 26 mumol/L/15 min) than in the evening (491 +/- 91 mumol/L/15 min). These results confirm diurnal variations in insulin sensitivity and plasma NEFA concentrations irrespective of feeding and exercise. We speculate that the relatively elevated plasma NEFA levels in the evening are the cause rather than the consequence of increased insulin resistance at this time. PMID- 10456208 TI - Correlation of marker values in Down's syndrome screening: the effect of dating error. AB - Central to any screening algorithm for Down's syndrome are the values used for the parameters in the multivariate Gaussian statistical model that is used to describe the joint distribution of the marker values. There has been much discussion about the values of the means and standard deviations which are appropriate but little interest has been shown in the values of the correlation coefficients between markers. There has been some speculation that the range of parameter values quoted in the literature arises from factors such as storage of samples, between-assay effects and differences in assay methodologies. We show that gestational dating error, among other factors, could be responsible for much of the variation that is present in quoted parameter values, though the factors mentioned above clearly have an effect. PMID- 10456209 TI - Are cotinine assays of value in predicting adverse pregnancy outcome? AB - We investigated the utility of maternal cotinine concentration as a predictor of preterm and small-for-gestational-age (SGA) deliveries. Subjects were randomly selected from antenatal clinics, and their serum cotinine concentrations at approximately 16 weeks' gestation were compared with self-reports of smoking. The value of these measures in predicting preterm and SGA deliveries was examined, using likelihood ratios and logistic regression, for 845 women with singleton, live births. Smoking status alone (defined by self-report or cotinine) only weakly predicted preterm or SGA delivery. Self-reported moderate/high smoking (top 15% of smokers) had a likelihood ratio (LR) of 2 and a positive predictive value (PPV) of 17%. In contrast, moderate/high cotinine concentration (top 15% of smokers) had an LR of 5 and a PPV of 33%. The predictive value of cotinine was independent of associated maternal characteristics. Cotinine concentration may therefore be a useful predictor of poor pregnancy outcome, and should be considered as a screening tool. PMID- 10456210 TI - Bloodspot testosterone assay suitable for study of neonates and monitoring of children with congenital adrenal hyperplasia. AB - A bloodspot assay for testosterone has been developed using an antiserum raised against testosterone-3-carboxymethyloxime-bovine serum albumin (T-3 CMO-BSA) conjugate and 125I-T-3CMO-histamine tracer. The method has a detection limit of 0.4 nmol/L with between-batch precision of less than 15% for testosterone values between 0.9 and 2.0 nmol/L and less than 10% for values greater than 2.0 nmol/L. Recovery of 4 nmol/L testosterone spiked into whole blood samples from 16 women (with unspiked bloodspot testosterone concentrations ranging from 0.4 to 3.7 nmol/L) was 100.9%. Bloodspot testosterone concentration was measured in thirty 6 9 day old infants (19 boys, 11 girls) with an absolute range in boys of < 0.4-2.3 nmol/L and in girls of < 0.4 nmol/L. In preterm infants (12 boys, 12 girls) of varying birth gestation and postnatal age, levels were higher than in term infants, with boys showing a rise with increasing postnatal age. Daytime bloodspot profiles in four children on replacement therapy for congenital adrenal hyperplasia demonstrated consistently low testosterone levels in the presence of moderately elevated early morning 17-hydroxyprogesterone concentrations, thus indicating adequate treatment. The bloodspot testosterone assay can be applied to the study of androgen physiology in preterm neonates as well as for monitoring treatment in disorders such as congenital adrenal hyperplasia. PMID- 10456211 TI - Clinical usefulness of biochemical resorption markers in osteoporosis. AB - We have evaluated three commercial assays for collagen cross-links, two urine assays and a recently developed serum assay, as markers of bone turnover in 30 postmenopausal women with osteoporosis during their first year of treatment with the anti-resorptive drug alendronate. Before treatment, urine free deoxypyridinoline crosslinks (Dpd), urine N-telopeptide crosslinks (NTx) and serum C-telopeptide (CTx) values were within postmenopausal reference ranges. After 3 months' treatment the decrease in NTx and CTx was greater than that of Dpd (-50%, P < 0.0001 and -48%, P < 0.0001, compared with -11%, NS), as it was after 6 months (-51%, P < 0.0001, and -57%, P < 0.0001, compared with -19%, P < 0.01). The decrease in resorption markers after 6 months was significant in 23% (Dpd), 66% (NTx) and 66% (CTx) of individuals. Neither baseline resorption marker values nor the per cent change after 6 months' therapy correlated with bone mineral density change (BMD) at either lumbar spine or femoral neck after one year's therapy, except baseline Dpd and lumbar spine BMD (P < 0.01). We conclude that NTx and serum CTx were more sensitive markers of bone turnover suppression by alendronate, but only baseline Dpd was useful in the prediction of individual bone density response after one year. PMID- 10456212 TI - Lymphocyte populations in tuberculous pleural effusions. AB - Different systemic and local responses to mycobacterial antigens suggest an active compartmentalization of responsive lymphocytes to tubercular antigens. This fact, observed in pleuritic processes, raises doubts about the accuracy of information obtained in the study of cells taken solely from peripheral blood. For this reason we decided to study the concept of compartmentalization in 140 patients suffering from pleural effusions. Patients were classified into six groups according to the aetiology of the effusion: group I, tuberculous, n = 23; group II, paraneoplastic, n = 41; group III, metapneumonic empyematous, n = 5; group IV, transudate, n = 38; group V, miscellaneous exudate, n = 19; group VI, unknown aetiology, n = 14. In each group we studied the lymphocyte population by using flow cytometry with doubly fluorescent monoclonal antibodies: B [expressing human lymphocyte antigen (HLA)-DR on the surface], T (CD3+), CD4+ and CD8+, and the subpopulation of activated T lymphocytes (together expressing CD3 and HLA-DR on the surface) (CD3+DR+). The study of these subpopulations in peripheral blood did not yield valuable results, but the CD3+DR+ population in pleural fluid demonstrated a diagnostic efficiency of 84% [positive predictive value (PPV) 51%, negative predictive value (NPV) 96%] at a cut-off value of 80.4 cells/mm3. The CD3+DR+ pleural fluid/peripheral blood ratio demonstrated an efficiency of 83% (PPV 50%, NPV 96%), and showed a statistically significant difference (P < 0.02) with regard to all the diagnostic groups, with the exception of the paraneoplastic effusions. The lymphocytic subpopulations study confirms the concept of compartmentalization in tuberculous pleuritis, as shown by the greater number of activated T lymphocytes present in pleural fluid in comparison with peripheral blood in tuberculous pleuritis, a 98% efficiency of adenosine deaminase (ADA) determination in pleural fluid versus a 50% value in peripheral blood, predominance of helper cells (CD4+) in pleural fluid and suppressor cells (CD8+) in peripheral blood, a greater CD4+/CD8+ ratio in pleural fluid than in peripheral blood, and a significant correlation of ADA-CD3+DR+ in pleural fluid, which does not occur in peripheral blood. PMID- 10456213 TI - 'Pseudohypouricosuria' in alcaptonuria: homogentisic acid interference in the measurement of urinary uric acid with the uricase-peroxidase reaction. AB - Urinary excretion of uric acid was found to be extremely low in a 58-year-old female patient with alcaptonuria. This was due to interference with the uricase peroxidase method used, because analysis using high-performance liquid chromatography (HPLC) showed a normal urinary concentration of uric acid. In vitro experiments demonstrated that a high concentration of homogentisic acid in the patient's urine inhibited the peroxidase reaction, possibly due to inhibition of the colour development of 3-methyl-N-ethyl-N-(beta-hydroxyethyl)aniline (MEHA) and 4-aminoantipyrine, via the peroxidase reaction. A homogentisic acid concentration equivalent to that in plasma did not affect the uricase-peroxidase reaction. This result suggests that any assay based on a peroxidase method is affected by a high urinary concentration of homogentisic acid in patients with alcaptonuria. PMID- 10456214 TI - Alleviation of carbon tetrachloride-induced chronic liver injury and related dysfunction by L-tryptophan in rats. AB - We examined whether L-tryptophan (Trp) alleviates carbon tetrachloride (CCl4) induced chronic liver injury and related dysfunction. Fifty rats were classified into four groups: the first (15 rats) served as the control; the second (10 rats) received subcutaneous injections of CCl4 (1.0 mL/kg) twice weekly for 8 weeks; the third (15 rats) received daily intraperitoneal injections of Trp (50 mg/kg) for 2 weeks after 6 weeks of CCl4 treatment; the fourth (10 rats) received both treatments. The activities of serum transaminases and the content of liver total hydroxyproline increased after 6 and 8 weeks of CCl4 treatment. The concentrations of serum albumin and liver protein and the in vitro activity of liver protein synthesis fell after 8 weeks of the treatment. Trp administration alleviated all these changes. At 6 and 8 weeks of CCl4 treatment the serum triglyceride concentration fell, whereas the liver triglyceride and lipid peroxide concentrations were elevated. Trp administration hardly affected these changes. These results indicate that Trp alleviates CCl4-induced chronic liver injury possibly by maintaining the activity of protein synthesis. PMID- 10456215 TI - Detection of p53 gene mutations in its full translational sequence by cleavase fragment length polymorphism analysis. PMID- 10456216 TI - Unusually slow-migrating albumin variants: rapid confirmation using immunofixation. PMID- 10456217 TI - Transient hyperphosphatasaemia in an adult with pre-existing liver disease. PMID- 10456218 TI - Hyperornithinaemia associated with gyrate atrophy of the choroid and retina: two cases from India. PMID- 10456219 TI - Metabolism of apolipoproteins AI and AII in a patient with paradoxical reduction in high-density lipoprotein due to ciprofibrate. PMID- 10456220 TI - Creatine kinase-BB isoenzyme in chest pain: an indicator that change in creatine kinase is not always due to an infarct? PMID- 10456221 TI - A case of factitious normocalcaemia. PMID- 10456222 TI - The portable laboratory and emergency department point-of-care testing. PMID- 10456223 TI - Review of investigation and management of severe hyponatraemia in a hospital population. PMID- 10456224 TI - Abbott IMx homocysteine assay: significant interference by 3-deazaadenosine. PMID- 10456225 TI - Sarco/endoplasmic reticulum Ca2+ (SERCA)-pumps: link to heart beats and calcium waves. AB - Mobilization of endoplasmic reticulum Ca2+ is pivotal to the ability of a cell to send or respond to stimuli. Ca(2+)-Mg(2+)-ATPases, termed SERCA pumps, sequester Ca2+ into the sarco/endoplasmic reticulum. There are several SERCA protein isoforms encoded by three genes. This paper summarizes the structure, function, tissue and subcellular distribution, and regulation of various SERCA isoforms. Then it attempts to link divergence in the signal transduction processes of cells to the types and levels of SERCA proteins they express and to how the cells regulate their SERCA pump activity. The paper examines possible linkages between SERCA pumps and receptor-activated Ca2+ entry, SERCA isoform localization and Ca(2+)-waves, and the role of SERCA pumps in nuclear Ca2+ in cell proliferation and apoptosis. Then it uses available information on cardiac function and chronic stimulation of the fast-twitch muscle to answer a series of basic questions on the regulation of SERCA activity and expression and their linkage to signal transduction. Finally, it discusses the possibility that neurons exhibit complex Ca(2+)-waves whose interactions have the potential to explain the operational basis of neural networks. A series of unanswered questions emerge based on this synthesis, including the unsettling issue of whether all the isoforms are needed to achieve the divergence in signal transduction or if there is a degree of redundancy in the system. PMID- 10456226 TI - Increase of human spermatozoa intracellular Ca2+ concentration after fusion with prostasomes. AB - Prostasomes are membranous vesicles (150-200 nm diameter) present in human semen. They are secreted by the prostate gland and contain large amounts of cholesterol, sphingomyelin and calcium, and some of their proteins are enzymes. Prostasomes are involved in a number of biological functions. In previous work, we discovered that prostasomes may fuse to sperm at neutral or at slightly acidic pH values. This mechanism may deliver calcium to sperm, thereby influencing sperm functions. We measured sperm [Ca2+]i with the fura-2 AM method and found that it increased after mixing prostasomes and sperm at pH values allowing fusion (pH 5-7). The increase of [Ca2+]i was proportional to the extent of fusion as measured through the relief of R18 self-quenching. We also examined the increase of sperm [Ca2+]i and the extent of fusion as a function of sperm to prostasome ratio and, also in this case, there was proportionality between the extent of fusion and the increase of [Ca2+]i that reached its maximal values in about 10-20 min. However, a detectable increase of [Ca2+]i was attained after 2 min of fusion. This would represent a new mechanism to influence sperm [Ca2+]i besides ion-exchange systems and ATP-dependent pumps. The value of [Ca2+]i remained elevated, unless Na+ was also present in the external medium. Therefore, the mechanism of fusion might influence deeply the physiology of sperm by producing a transient increase of [Ca2+]i. PMID- 10456227 TI - Interplay between Ca2+ release and Ca2+ influx underlies localized hyperpolarization-induced [Ca2+]i waves in prostatic cells. AB - Calcium seems to be a major second messenger involved in the regulation of prostatic cell functions, but the mechanisms underlying its control are poorly understood. We investigated spatiotemporal aspects of Ca2+ signals in the LNCaP cell line, a model of androgen-dependent prostatic cells, by using non-invasive external electric field pulses that hyperpolarize the anode facing membrane and depolarize the membrane facing the cathode. Using high-speed fluo-3 confocal imaging, we found that an electric field pulse (10-15 V/cm, 1-5 mA, 5 ms) initiated rapidly, at the hyperpolarized end of the cell, a propagated [Ca2+]i wave which spread through the cell with a constant amplitude and an average velocity of about 20 microns/s. As evidenced by the total wave inhibition either by the block of Ca2+ entry or the depletion of Ca2+ stores by thapsigargin, a specific Ca(2+)-ATPase inhibitor, the [Ca2+]i wave initiation may imply a localized Ca2+ influx linked to a focal auto-regenerative process of Ca2+ release. Using different external Ca2+ and Ca2+ entry blockers concentrations, Mn2+ quenching of fluo-3 and fura-2 fluorescence and inhibitors of InsP3 production, we found evidence that the [Ca2+]i wave progression required, in the presence of basal levels of InsP3, an interplay between Ca2+ release from InsP3 sensitive Ca2+ stores and Ca2+ influx through channels possibly activated by the [Ca2+]i rise. PMID- 10456229 TI - Inhibition of Na/Ca exchange by external Ni in guinea-pig ventricular myocytes at 37 degrees C, dialysed internally with cAMP-free and cAMP-containing solutions. AB - In many mammalian tissue types an integral membrane protein--the sodium/calcium (Na/Ca) exchanger--plays a key role in intracellular Ca homeostasis, and evidence suggests that Na/Ca exchange function can be modulated by cAMP-dependent phosphorylation. External Nickel (Ni) ions are used widely to inhibit the exchange but little is known about the mode of Ni action. In guinea-pig ventricular myocytes, we investigated inhibition of Na/Ca exchange by external Ni under phosphorylated (cells dialysed with cAMP) and non-phosphorylated conditions. Ventricular myocytes were isolated from adult guinea-pig hearts, recordings were made at 37 degrees C using the whole-cell patch clamp technique. Internal and external solutions were used which allowed Na/Ca exchange current (INaCa) to be measured during a descending voltage ramp protocol (+80 to -120 mV) applied from a holding potential of -40 mV. The application of 10 mM Ni caused a maximal block of INaCa since inhibition was identical to that when a Na- and Ca free (0Na/0Ca) solution was superfused externally. Kinetics of Ni-block of INaCa were assessed using applications of different external [Ni] to cells dialysed internally with cAMP-free and 100 microM cAMP-containing solutions. At +60 mV, Ni inhibited INaCa in cells dialysed with a cAMP-free solution with a dissociation constant (KD) of 0.29 +/- 0.03 mM and the data were fitted with a Hill coefficient of 0.89 +/- 0.07 (n = 9 cells). In cells dialysed with 100 microM cAMP the exchange was inhibited by Ni with a KD of 0.16 +/- 0.05 mM, the Hill coefficient was 0.82 +/- 0.16 (n = 6-7 cells). The KD and Hill coefficient values obtained in cells dialysed with cAMP-free and cAMP-containing solutions were not significantly different. Inhibition of INaCa by Ni did not appear to be voltage dependent, was maximal within 3-4 s of application and was rapidly reversible. With cAMP-free internal dialysate, inhibition was 'mixed' showing competition with external Ca and a degree of non-competitive block. With 100 microM cAMP the inhibition appeared to be more non-competitive. We conclude that, under these experimental conditions, a concentration of external Ni of 10 mM is sufficient to produce maximal inhibition of INaCa in guinea-pig cardiac cells. PMID- 10456228 TI - Ca2+ sensitivity of phospholipid scrambling in human red cell ghosts. AB - The phospholipids in plasma membranes of erythrocytes, as well as platelets, lymphocytes and other cells are asymmetrically distributed, with sphingomyelin and phosphatidylcholine residing predominantly in the outer leaflet of the bilayer, and phosphatidylserine and phosphatidylethanolamine in the inner leaflet. It is known that Ca2+ can disrupt the phospholipid asymmetry by activation of a protein known as phospholipid scramblase, which affects bidirectional phospholipid movement in a largely non-selective manner. As Ca2+ also inhibits aminophospholipid translocase, whose Mg(2+)-ATPase activity is responsible for active translocation of aminophospholipids from the outer to the inner leaflet, it is important to accurately determine the sensitivity of scramblase to intracellular free Ca2+. In the present study we have utilized the favourable Kd of Mag-fura-2 for calcium in the high micromolar range to determine free Ca2+ levels associated with lipid scrambling in resealed human red cell ghosts. The Ca2+ sensitivity was measured in parallel to the translocation of a fluorescent-labelled lipid incorporated into the ghost bilayer. The phospholipid scrambling was found to be half-maximally activated at 63-88 microM free intracellular Ca2+. The wider applicability of the method and the physiological implications of the calcium sensitivity determined is discussed. PMID- 10456230 TI - An electrogenic ionic pump derived from an ionotropic receptor: assessment of a candidate. AB - 1. Data obtained studying permeability characteristics of single Deiters' membranes in a microchamber system show that intracellular GABA can activate chloride in-->out passage with a GABAA pharmacology. 2. The overall data suggest the presence of a chloride extrusion pump in these neurons based on intracellular GABA activated chloride channels. 3. This conclusion takes up a previous theoretical suggestion that ionic channels could work as ionic pumps provided an energy input modifies the energy profile along the permeation path. 4. According to our quantitative evaluation, this pumping mechanism works with a low yield and along a cycle with a strongly asymmetric behavior, being far from equilibrium due to powerful "leakage" pathways for chloride in these neurons. PMID- 10456231 TI - Effects of diethylstilbestrol on mouse hippocampal evoked potentials in vitro. AB - 1. Several steroids and related compounds can bind to central opiate receptors in whole-brain mouse homogenates. Among these drugs, the synthetic estrogen, diethylstilbestrol (DES), exhibits one of the highest affinities in binding experiments labeling opiate receptors with the nonselective opiate antagonist, [3H]diprenorphine. 2. In the search for a functional correlate to this biochemical finding, we have studied the effects of DES on the mouse hippocampal slice in vitro preparation. 3. Previously, binding studies were performed in hippocampal homogenates, labeling opiate receptors with [3H]diprenorphine or with the mu-selective opiate agonist, [3H]DAGO. DES inhibited [3H]diprenorphine and [3H]DAGO binding, the IC50 values obtained being (1.03 +/- 0.16) x 10(-5) and (1 +/- 0.8) x 10(-5) M, respectively. 4. In mice hippocampal slices, we measured the extracellular evoked potentials obtained in the CA1 pyramidal cell layer of the hippocampi and the field excitatory postsynaptic potentials (EPSP) obtained in the stratum radiatum. The presence of DES (10(-5) M) induced an increase in the amplitude of the population spikes measured in the pyramidal layer without modifying the field EPSP. This effect is similar to that obtained in the presence of DAGO in this preparation. The effect produced by DES was not modified by the presence of the opiate competitive antagonist, naloxone (10(-5) M), or by the opiate alkylating agent, beta-chlornaltrexamine (10(-5) M). Conversely, in the presence of the transcription inhibitor, actinomycin D (5 micrograms/ml), the effect produced by DES was inhibited. 5. Our results with DES support the general idea that estrogens increase central excitability. Although diethylstilbestrol can bind to opiate receptors in the hippocampus, the effect induced by this estrogen on hippocampal excitability seems unrelated to a direct action on opiate receptors, and an intracellular effect is suggested. PMID- 10456232 TI - Glutamate receptor requirement for neuronal death from anoxia-reoxygenation: an in Vitro model for assessment of the neuroprotective effects of estrogens. AB - 1. Previous studies demonstrated that estrogens, specifically 17 beta-estradiol, the potent, naturally occurring estrogen, are neuroprotective in a variety of models including glutamate toxicity. The aim of the present study is twofold: (1) to assess the requirement for glutamate receptors in neuronal cell death associated with anoxia-reoxygenation in three cell types, SK-N-SH and HT-22 neuronal cell lines and primary rat cortical neuronal cultures, and (2) to evaluate the neuroprotective activity of both 17 beta-estradiol and its weaker isomer, 17 alpha-estradiol, in both anoxia-reoxygenation and glutamate toxicity. 2. SK-N-SH and HT-22 cell lines, both of which lack NMDA receptors as assessed by MK-801 binding assays, were resistant to both anoxia-reoxygenation and glutamate induced cell death. In contrast, primary rat cortical neurons, which exhibit both NMDA and AMPA receptors, were sensitive to brief periods of exposure to anoxia reoxygenation or glutamate. As such, there appears to be an obligatory requirement for NMDA and/or AMPA receptors in neuronal cell death resulting from brief periods of anoxia followed by reoxygenation. 3. Using primary rat cortical neuronal cultures, we evaluated the neuroprotective activity of 17 beta-estradiol (1.3 or 133 nM) and 17 alpha-estradiol (133 nM) in both anoxia-reoxygenation and excitotoxicity models of cell death. We found that the 133 nM but not the 1.3 nM dose of the potent estrogen, 17 beta-estradiol, protected 58.0, 57.5, and 85.3% of the primary rat cortical neurons from anoxia-reoxygenation, glutamate, or AMPA toxicity, respectively, and the 133 nM dose of the weak estrogen, 17 alpha estradiol, protected 74.6, 81.7, and 85.8% of cells from anoxia-reoxygenation, glutamate, or AMPA toxicity, respectively. These data demonstrate that pretreatment with estrogens can attenuate glutamate excitotoxicity and that this protection is independent of the ability of the steroid to bind the estrogen receptor. PMID- 10456233 TI - Chronic effects of xanthines on levels of central receptors in mice. AB - 1. Chronic ingestion of caffeine causes a significant increase in levels of A1 adenosine, nicotinic and muscarinic receptors, serotonergic receptors, GABAA receptors and L-type calcium channels in cerebral cortical membranes from mice NIH Swiss strain mice. 2. Chronic theophylline and paraxanthine had effects similar to those of caffeine except that levels of L-type channels were unchanged. Chronic theobromine, a weak adenosine antagonist, and 1-isobutyl-3 methylxanthine (IBMX), a potent adenosine antagonist and phosphodiesterase inhibitor, caused only an increase in levels of A1-adenosine receptors. A combination of chronic caffeine and IBMX had the same effects on receptors as caffeine alone. Chronic 3,7-dimethyl-1-propargylxanthine (DMPX), a somewhat selective A2A-antagonist, caused only an increase in levels of A1-adenosine receptors. Pentoxifylline, an adenosine-uptake inhibitor inactive at adenosine receptors, had no effect on receptor levels or calcium channels. 3. A comparison of plasma and brain levels of xanthines indicated that caffeine penetrated more readily and attained somewhat higher brain levels than theophylline or theobromine. Penetration and levels were even lower for IBMX, paraxanthine, DMPX, and pentoxyfylline. 4. The results suggest that effective blockade of both A1 and A2A-adenosine receptors is necessary for the full spectrum of biochemical changes elicited by chronic ingestion of xanthines, such as caffeine, theophylline, and paraxanthine. PMID- 10456234 TI - Cotransport of glyceraldehyde-3-phosphate dehydrogenase and actin in axons of chicken motoneurons. AB - 1. To study proteins transported with actin in axons, we pulse-labeled motoneurons in the chicken sciatic nerve with [35S]methionine and, 1-20 days later, isolated actin and its binding proteins by affinity chromatography of Triton soluble nerve extracts on DNase I-Sepharose. The DNase I-purified proteins were electrophoresed on two-dimensional gels and the specific activity of the radioactively labeled protein spots was estimated by fluorography. 2. In addition to actin, which binds specifically to DNase I, a small number of other proteins were labeled, including established actin monomer binding proteins and a protein of 36 kDa and pI 8.5. On the basis of its molecular mass, pI, amino acid composition, and immunostaining, the unrecognized protein was identified as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). 3. The high affinity binding of GAPDH to actin was confirmed by incubation of Triton-soluble nerve extracts with either mouse anti-GAPDH (or antiactin) and indirect immunomagnetic separation with Dynabeads covalently linked to sheep anti-mouse antibody. Analysis by one-dimensional gel electrophoresis and immunoblotting showed that actin and GAPDH were the main proteins isolated by these methods. 4. Analysis of labeled nerves at 12 and 20 days after pulse labeling showed that GAPDH and actin were transported at the same rate, i.e., 3-5 mm/day, which corresponds to slow component b of axonal transport. These proteins were not associated with rapidly transported proteins that accumulated proximal to a ligation 7 cm from the spinal cord 9 hr after injection of radioactivity. 5. Our results indicate that GAPDH and actin are transported as a complex in axons and raise the possibility that GAPDH could act as a chaperone for monomeric actin, translocating it to intraaxonal sites for exchange with or assembly into actin filaments. Alternatively, actin could be involved in translocating and anchoring GAPDH to specialized sites in axons and nerve terminals that require a source of ATP by glycolysis. PMID- 10456236 TI - A rabbit model of proliferative vitreoretinopathy induced by injection of astrocytic cultures. AB - 1. The objective of this study was to decipher whether proliferation of astrocytes and invasion of astrocytic processes into the retina could contribute to retinal detachment in a rabbit model. 2. Cultures of astrocytes were injected intravitreally into the eyes of albino rabbits. 3. Two weeks after injection, proliferation of astrocytes on the retinal surfaces was observed. Vascular endothelial growth factor (VEGF) and proliferative cell nuclear antigen (PCNA) were found by immunohistochemistry to be expressed in the center of the astrocytic growth. 4. Using the same immunohistochemical technique to visualize glial fibrillary acidic protein (GFAP), a marker for astrocytes, processes of astrocytes in the growth were observed to penetrate into the host retina. 5. Retinal detachment was then confirmed by ultrasound, histologically, and grossly 2 weeks after injection of astrocytes. 6. Histochemistry on esterase indicated chloroesterase positive cells inside the growth. The secretion of this form of esterase might soften the vitreous and enhanced retinal detachment. 7. Six weeks after injection, VEGF and PCNA decreased in the astrocytic growth but astrocytic processes still attached onto and penetrated the host retina. 8. This study suggests that astrocytes could be a major factor in inducing retinal detachment. PMID- 10456235 TI - Determinants of phencyclidine potency on the nicotinic acetylcholine receptors from muscle and electric organ. AB - 1. Phencyclidine (PCP) is an inhibitor of the nicotinic acetylcholine receptor (AChR) with characteristics of an open-channel blocker. The location of PCP binding site on the AChR molecule is unknown. 2. PCP inhibits the AChR from electric organ with a higher potency than muscle AChR. To find the molecular basis of this difference, we expressed the two native and six hybrid receptors, and two receptors containing mutated mouse gamma subunits in Xenopus laevis oocytes. The inhibition of ACh-induced current in these receptors by PCP was studied using whole-cell voltage-clamp. All hybrid receptors generated robust ACh induced currents, while incomplete receptors (gamma-less or delta-less) did not. 3. PCP potency was higher on hybrids containing Torpedo beta and gamma subunits regardless of the alpha and delta subunit origin. A mouse gamma subunit containing the asparagine 6' to the serine mutation in the M2 segment conferred a high sensitivity to PCP. 4. These results support the conclusion that the amino acid residues at the position 6' of the M2 segments contribute to the PCP potency difference between Torpedo and mouse receptors. 5. Another noncompetitive inhibitor of the AChR, the cembranoid eupalmerin acetate (EUAC), also inhibited the electric organ receptor with a somewhat higher potency than muscle AChR. However, the IC50 values for EUAC inhibition of hybrid receptors did not follow the pattern observed for PCP. Therefore, these two inhibitors interact differently with the AChR molecule. PMID- 10456237 TI - The influence of gender and age on neonatal rat hypothalamic 5-HT1A and 5-HT2A receptors. AB - 1. Rat hypothalamic 5-hydroxytryptamine (5-HT) and 5-hydroxyindole acetic acid (5 HIAA) concentrations are transiently sexually differentiated in the second week postpartum (pp), with higher levels in the female. In this report we investigate the possibility that 5-HT receptors may also exhibit sexual dimorphism in the neonatal period. 2. 5-HT1A and 5-HT2A receptors were quantitated by radioligand binding of [3H]ketanserin and [3H]8-OH DPAT, respectively, in hypothalamus and amygdala from male and female rats at days 8-16 pp. 3. There was no sexual dimorphism or change in the density of 5-HT2A binding in hypothalamus or amygdala over days 8-16 pp. There was also no sexual dimorphism of 5-HT1A receptors. 4. There was an increase in 5-HT1A receptor density in both the hypothalamus and the amygdala. In the hypothalamus, but not the amygdala, this increase was interrupted on day 14 by a decrease in 5-HT1A receptors, which we suggest may be of physiological significance in modifying the eventual pattern of adult agonistic activity. 5. The results suggest that the sexual dimorphism in 5-HT turnover is predominantly presynaptic, relating to altered synthesis and/or release, and is not of sufficient magnitude or duration to produce adaptive responses in postsynaptic 5-HT1A or 5-HT2A receptors. PMID- 10456238 TI - Down syndrome: imaging of multiorgan involvement. AB - Down syndrome (trisomy 21) has many manifestations that affect multiple organ systems, and we describe the wide array of imaging findings. Common cardiovascular and gastrointestinal entities are congenital heart disease (atrioventricular canal), bowel atresias (duodenal and anal), and Hirschsprungs disease. Children with Down syndrome have an 18-20 fold increased incidence of leukemia. Pulmonary hypoplasia, lung cysts, and pig bronchus (origin of the right upper lobe bronchus from the trachea) have been described. Neurologic findings include mineralizing vasculopathy of the basal ganglia, Moyamoya disease, and cerebellar/vermian hypoplasia. Musculoskeletal manifestations are numerous and include eleven ribs, hypersegmented sternum, abnormal pelvis, joint laxity/dislocations, and DDH (developmental dysplasia of the hip). Of special importance is the "triple jeopardy" of the upper cervical spine (atlanoaxial subluxation, hypoplastic posterior arch of C1, and atlantooccipital instability) and the resulting controversial cervical spine radiographic screening of children with Down syndrome. Knowledge of the many anomalies associated with Down syndrome can aid the clinician, not only in diagnosing abnormalities in these patients, but also in counseling families for potential problems that can occur in these children. PMID- 10456239 TI - Feeding, medical factors, and developmental outcome in premature infants. AB - This is a prospective, longitudinal study of premature infants investigating whether the length of time needed to reach full enteral feedings (FEF) or full nipple feedings (FNF) is related to medical complications and/or developmental outcome at 24 months corrected age. Premature infants (n = 161) from three institutions with birth weights less than 1,600 grams were followed up from birth to 24 months corrected age. The infants were stratified into groups by the severity of medical complications. Bayley Scales of Infant Development were performed at 24 months corrected age. Multiple linear regression was used to analyze the association between feeding milestones, medical complications, and developmental outcomes. Our results show that when controlling for birth weight and gestational age (GA), the severity of respiratory complications was significantly related to reaching FEF (p = 0.024) and FNF (p = 0.0014). Furthermore, when controlling for the severity of respiratory complications, GA, and socioeconomic status, an increased length of time to FEF was significantly associated with a poorer mental outcome (p = 0.0013). We conclude that there is an association between the length of time to reach FEF and mental developmental outcome at 24 months corrected age. Infants who reach full enteral feedings at an earlier age appear to have a better developmental outcome despite their GA and severity of respiratory complications. PMID- 10456240 TI - Low frequency of meperidine-associated seizures in sickle cell disease. AB - Pain control measures in sickle cell diseases are not uniform. Most clinicians use parenteral morphine or meperidine for severe pain. Reports of seizures associated with meperidine have led to a growing reliance on intravenous morphine, usually with patient-controlled devices. Acceptance of morphine has been poor among patients, and many prefer meperidine. The aim of this retrospective study was to determine the incidence of meperidine-associated seizures in a large, mostly pediatric population with sickling disorders. The results suggest that the incidence of seizures with meperidine is extremely small (0.4% of patients; 0.06% of admissions). The risk of seizures should not dissuade clinicians from using this drug. PMID- 10456241 TI - Comparison of ear to rectal temperature measurements in infants and toddlers. AB - The purpose of this study was to compare arterial heat balance ear temperature measurements to rectal temperatures in infants and children and to determine the ability of the ear thermometer being tested to detect fever. From 12/95 to 2/96, 1,175 pairs of ear and rectal temperature measurements were prospectively obtained from 140 infants and toddlers. The mean rectal temperature was 37.58 degrees C (sd = 0.68) and the mean ear temperature was 37.60 degrees C (sd = 0.85). However, at the low end of the rectal temperature scale, ear temperatures tended to be higher, and at the high end of the rectal temperature scale, ear temperatures tended to be lower. There were 292 readings with a rectal temperature > or = 38.0 degrees C and in 204 (70%) the ear temperature was also > or = 38.0 degrees C. A retrospective analysis of 53 children who became febrile in hospital (ear or rectal temperature > or = 38.0 degrees C) showed that fever was detected first by rectal measurement in seven, by ear measurement in 31 (59%), and by both in 15 (28%). These data indicate that, on the average, rectal and ear temperature measurements are not different. Fever that developed in children after hospitalization was more likely to be first detected by ear than by rectal measurement. PMID- 10456242 TI - Effects of skin-to-skin holding on general movements of preterm infants. AB - The study objective was to test the hypothesis that the effect of skin-to-skin (STS) holding increases the ratio of rest to activity in low birth weight preterm infants. Ten infants with birthweight < 2,000 grams were videotaped before and after STS holding. Video recordings were analyzed to determine the number of general movements. We found no statistically significant difference between the percentage of general movements over the two periods. We conclude that the ratio of rest-activity before and after STS holding does not change as measured by occurrence of general movements. PMID- 10456243 TI - Intention to practice primary care by pediatric residents: nature or nurture? AB - Factors associated with the intention to practice primary care were examined in a survey of a national sample of PL-2 residents (n = 98). Socioemotional orientation (nature), faculty and peer encouragement (nurture), and clinical experiences during residency (nurture) were independently associated with a primary care career choice. For residents who changed career intentions to primary care from a nonprimary care preference, gender, encouragement by faculty and peers, and outpatient experiences during residency were associated with the change. Encouragement by both faculty and peers had the strongest influence on primary care career choice for all residents. PMID- 10456245 TI - Group A streptococcus and the pediatrician: old and new. PMID- 10456244 TI - Group A streptococcal multifocal septic arthritis: a case report. PMID- 10456246 TI - Nonobstructive cor triatriatum in infancy. PMID- 10456247 TI - Test ordering on children with acute abdominal pain. PMID- 10456248 TI - Glucocorticoids and muscle catabolism. AB - Glucocorticoids inhibit protein synthesis and stimulate protein degradation in skeletal muscle and are an important factor in the development of muscle atrophy in various catabolic conditions. Glucocorticoid-stimulated muscle protein breakdown is primarily caused by ubiquitin-proteasome-dependent proteolysis although calcium-dependent protein degradation may also be involved. In certain catabolic conditions, including sepsis, an interaction between glucocorticoids and proinflammatory cytokines is important for the stimulation of muscle protein breakdown. PMID- 10456249 TI - Amino acid-dependent signal transduction and insulin sensitivity. AB - Recent developments indicate that amino acids, in addition to their function as substrates for many metabolic pathways, can stimulate a signal transduction pathway that shares components with insulin-stimulated signalling cascades. Insulin sensitivity is dependent on the ambient amino acid concentration. Amino acid-dependent signal transduction is present in all insulin-sensitive tissues and in pancreatic beta cells. A defect in amino acid-dependent signal transduction may result in phenomena similar to those found in diabetes mellitus. PMID- 10456250 TI - Potential intracellular targets for anabolic/anti-catabolic therapies. AB - The intracellular signalling pathways controlling muscle protein synthesis and proteolysis are potential targets for anabolic/anti-catabolic therapy. In this review, we consider both the potentiation of the effect of anabolic hormones and suppression of the catabolic action of cytokines. Potential candidates, in particular isoforms of the protein kinase C family, and their role in the control of ribosomal action and the ubiquitin-proteasome proteolytic system are discussed. PMID- 10456251 TI - The anti-catabolic effects of n-3 fatty acids. AB - The reversal of catabolic processes remains a significant challenge, partly related to the complexity of such processes and our incomplete understanding of the mechanisms involved. The n-3 fatty acids may be able to attenuate catabolism. This review examines the potential sites of action for these fatty acids and the evidence that supports their anti-catabolic properties. PMID- 10456252 TI - Cysteine and glutathione in catabolic conditions and immunological dysfunction. AB - The conspicuous increase in the plasma cysteine disulphide/thiol ratio in elderly persons and cancer patients indicates a shift of the plasma redox state. The most important redox buffers in skeletal muscle tissue and blood plasma, i.e. glutathione and albumin, respectively, are significantly decreased in different models of cachexia. Treatment with N-acetyl cysteine, i.e. a thiol-containing antioxidant, was found to increase the plasma albumin level and to ameliorate the loss of body cell mass in cancer patients and healthy individuals. The treatment of HIV infection with N-acetyl cysteine, in contrast, serves mainly as a tool to ameliorate the physiological and immunological consequences of the virus-induced cysteine deficiency. PMID- 10456253 TI - Inter-organ protein and carbohydrate metabolic relationships during sepsis: necessary evils or uncanny coincidences? AB - Sepsis alters the dynamic flux of metabolic substrates between skeletal muscle and liver. Derangements in skeletal muscle glucose metabolism evoked by sepsis to a certain extent determine the rate of gluconeogenesis in the liver. In contrast, accelerated rates of gluconeogenesis do not drive net catabolism in skeletal muscle, nor does the upregulation of hepatic protein metabolism in sepsis or inflammation appear to be contingent or dependent upon the catabolism of muscle proteins during sepsis. PMID- 10456254 TI - Anabolic steroids. AB - Anabolic steroids may be an additional mode of intervention to promote anabolism and improve clinical outcome in various acute and chronic wasting diseases. The present review discusses the rationale for anabolic steroid treatment in acute and chronic disease, their mechanistic actions, the available clinical trials in acute and chronic disease and their side-effects. PMID- 10456255 TI - Bibliography. Current world literature. Anabolic and catabolic signals. PMID- 10456257 TI - Immunopathogenesis of crescentic glomerulonephritis. AB - Crescentic glomerulonephritis provides an important therapeutic challenge because of its rapidly progressive course and poor outcome. Studies in animal models have elucidated some of the pivotal pathogenetic mechanisms, and human studies increasingly support the clinical relevance of these animal data. Accumulating evidence suggests that crescentic glomerulonephritis results from a complex cell mediated nephritogenic immune response. Interruption of a number of immune and inflammatory mediators can improve the outcome of this disease. PMID- 10456258 TI - New insights into the pathogenesis of interstitial nephritis. AB - Emerging data on the role of interstitial inflammation in progressive renal disease are redefining our understanding of the pathogenic mechanisms of chronic kidney damage. Recent experimental evidence emphasizes the role for both immune and non-immune mechanisms in interstitial nephritis. New observations regarding maneuvers that downregulate such injurious renal responses may direct further studies that develop new therapeutic modalities. PMID- 10456259 TI - Glomerular inflammation: use of genetically deficient mice to elucidate the roles of leukocyte adhesion molecules and Fc-gamma receptors in vivo. AB - Single gene knock-outs in mice have been used to define the biological role of leukocyte adhesion receptors, Fc-gamma receptors and complement in animal models of immune complex glomerulonephritis. These studies have shown important differences in the role of P-selectin in glomerular inflammation and inflammation at other sites, and have given a new appreciation of the dominant role played by Fc-gamma receptors in immune complex-induced glomerular injury. PMID- 10456260 TI - Role of nucleosomes for induction and glomerular binding of autoantibodies in lupus nephritis. AB - The cardinal feature of systemic lupus erythematosus is the formation of anti nuclear antibodies. In recent years, it has become clear that the nucleosome is a major autoantigen that drives this T cell-dependent autoimmune response, as exemplified by the presence of nucleosome-specific T helper cells and the high prevalence of nucleosome-specific autoantibodies. The only way to generate nucleosomes in vivo is by the process of apoptosis. There is growing evidence that in systemic lupus erythematosus apoptosis is disturbed, leading to the release of nucleosomes. Moreover, apoptosis-induced modifications of these autoantigens may render them more immunogenic, especially if the removal of apoptotic cells is insufficient. The first indications for the impaired clearance of apoptotic cells in systemic lupus erythematosus are emerging. Nucleosomes are also important for mediating tissue lesions, especially glomerulonephritis. In lupus nephritis nucleosomes, nucleosome-specific antibodies and nucleosome/IgG complexes have been identified in the glomerular immune deposits. Via their cationic histone part nucleosomes mediate the binding of anti-nuclear antibodies to intrinsic constituents of the glomerular basement membrane, such as the anionic heparan sulfate and collagen IV. Appreciation of this binding mechanism may lead to new treatment strategies, as shown for non-coagulant heparinoids. PMID- 10456261 TI - What is new with anti-neutrophil cytoplasmic antibodies: diagnostic, pathogenetic and therapeutic implications. AB - Anti-neutrophil cytoplasmic antibodies, as detected by indirect immunofluorescence, have limited diagnostic significance as they occur in a variety of inflammatory disorders. The presence of antibodies to defined target antigens of anti-neutrophil cytoplasmic antibodies, that is proteinase 3 and myeloperoxidase, is, however, highly specific for one of the systemic vasculitides, in particular Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome, and idiopathic pauci-immune necrotizing crescentic glomerulonephritis. In general, anti-proteinase-3-positive patients show more widespread organ involvement, more granuloma formation, and a more severe initial course of their renal lesions than anti-myeloperoxidase-positive patients; however, there is considerable overlap, and either antibody specificity may be found in the different clinical syndromes. In vitro, anti-neutrophil cytoplasmic antibodies are able further to activate pre-activated neutrophils and monocytes, which can result in endothelial damage. A direct activating effect of anti neutrophil cytoplasmic antibodies on endothelial cells has been suggested, but those studies should be confirmed. In vivo, experimental data support a pathogenetic role for anti-neutrophil cytoplasmic antibodies, particularly anti myeloperoxidase, but besides anti-neutrophil cytoplasmic antibodies a second pro inflammatory stimulus seems to be required to induce lesions. Whether anti neutrophil cytoplasmic antibodies can be a direct target for treatment has still to be proved. Current immunosuppressive treatment regimens for the anti neutrophil cytoplasmic antibody-associated vasculitides are, however, unsatisfactory because of side-effects, that is opportunistic infections and malignancies. New treatment regimens, based on new pathogenetic concepts, are currently being tested. PMID- 10456262 TI - Current trends in transplant immunology. AB - Recent advances in transplant immunology are wide ranging. These include the testing of new approaches to tolerance induction by the interruption of co stimulatory pathways, the analysis of molecular events underlying the development of chronic rejection, efforts to increase the donor pool by consideration of the role of brain death in donor-dependent outcomes of allografting, and progress towards renal xenografting. In addition, current molecular approaches are paramount to understanding key events from ischemia/reperfusion injury and the role of apoptosis in remodelling of the host immune response to an allograft. Important papers relevant to the field from the past year are reviewed with an eye to clinically relevant new data involving renal transplantation. PMID- 10456263 TI - Advances in the molecular basis of renal neoplasia. AB - The past 2 years have provided exciting progress in elucidating the molecular basis of renal cancer. Work on the von Hippel-Lindau tumor suppressor, pVHL, in clear-cell renal cancer is already suggesting new potential therapies, and should have important implications in the pathogenesis of renal cystic disease and tumor angiogenesis. In addition, study of the Wilms' tumor suppressor, WT1, is revealing much about the pathogenesis of Wilms' tumor, urogenital development, and glomerular podocyte biology. c-met, the gene encoding the hepatocyte growth factor receptor, has recently been identified as a causative gene for hereditary papillary renal cancer. This review will highlight these and other new molecular advances in the renal cancer field. PMID- 10456264 TI - Angiotensin converting enzyme inhibitors: first line therapy in patients with diabetic hypertension? PMID- 10456266 TI - Weight change, ideal weight and hypertension. AB - A growing body of evidence supports the fact that body weight is related to blood pressure. The risk of hypertension increases with increasing body mass index and weight gain; and weight loss reduces blood pressure. Birthweight may also be linked to adult blood pressure, possibly as a result of its relationship with later obesity. PMID- 10456265 TI - Pulse pressure, arterial compliance and cardiovascular morbidity and mortality. AB - Pulse pressure, an indicator of conduit vessel stiffness, has recently emerged as an important predictor of adverse cardiovascular events. The normally compliant conduit vessels provide an essential buffering role that minimizes potentially harmful pressure swings associated with intermittent pumping by the heart. A number of common conditions and acknowledged cardiac risk factors stiffen the conduit vessels, resulting in a vicious cycle of progressive vessel stiffening, increasing pulsatile load, pressure-related end-organ damage and clinical events. An emerging awareness of the central role of conduit vessel stiffness in cardiovascular pathophysiology has important implications for the classification and treatment of hypertension and other disorders that affect conduit vessel function and pulsatile hemodynamics. PMID- 10456267 TI - Blood pressure and cognitive decline in the elderly. AB - Elevated blood pressure is associated with cognitive decline in elderly people. Classically, hypertension was thought to lead to end-organ damage of the brain manifested by neuropsychological deficits. This review examines recent evidence for this hypothesis and also considers other possible causal mechanisms for the observed relationship between blood pressure and mental ability. PMID- 10456268 TI - Alcohol intake and cardiovascular morbidity and mortality. AB - Recent studies reinforce the consistent finding of a J-shaped inverse association between alcohol and cardiovascular disease morbidity and mortality, primarily as a result of the underlying association between alcohol and coronary heart disease. The most plausible mechanism behind this effect is through increases in high-density lipoprotein cholesterol. All types of alcoholic beverage reduce the risk of cardiovascular disease, suggesting that ethanol is responsible for the protective effect. PMID- 10456269 TI - Microalbuminuria in essential hypertension. AB - Urinary excretion of albumin exceeds normal values in 10 to 25% of patients with essential hypertension. The level of albuminuria is highly correlated with arterial pressure, and more closely with ambulatory arterial pressure. The interaction between albuminuria and arterial pressure is enhanced by overweight, smoking, protein intake, insulin resistance, lipid abnormalities, and possibly genotypes of the components of the renin-angiotensin system. The renal mechanisms of microalbuminuria are not well elucidated. Notably, an increase in filtration fraction suggestive of intraglomerular hypertension was observed in patients with hyperfiltration. Microalbuminuria may be a marker of diffuse vascular abnormalities predisposing to cardiovascular disease and/or hypertensive renal disease heralding future renal failure, but its predictive value needs to be tested in more long-term follow-up studies. Antihypertensive treatment has a varied influence on albuminuria; angiotensin-converting enzyme inhibitors may correct this abnormality (at least partially) better than other agents. PMID- 10456270 TI - The epidemic of cardiovascular disease in end-stage renal disease. PMID- 10456272 TI - Epidemiology and prevention. PMID- 10456271 TI - Renal immunology and pathology. PMID- 10456273 TI - Long-term results of organ preservation in chondrosarcoma of the cricoid. AB - Chondrosarcoma of the larynx is rare. The posterolateral lamina of the cricoid cartilage is the site most commonly involved. Although the symptomology, radiological and histopathological diagnostic features are well known, the condition continues to be diagnosed late and recurrences after excision are common. In general, radical surgical excision can result in long-term remission, although prognosis is related to the histopathological grade of the tumor. While a conservative surgical approach in low-grade tumors is justified, unexplained hoarseness, unilateral vocal cord paralysis and dyspnea requires CT scan examination. PMID- 10456274 TI - Changing trends in the occurrence and subsite distribution of laryngeal cancer in Finland. AB - Cancer of the larynx represents worldwide approximately 1-2% of all cancers and generally occurs predominantly in males. Based upon many reports, the age adjusted incidence of laryngeal cancer has been rising in recent years, especially among women. Squamous cell carcinomas arising in the glottic region are the most common of all laryngeal cancers and more prevalent than the supraglottic ones. Subglottic disease is still rare. The Finnish Head and Neck Oncology Group evaluated the present occurrence of laryngeal cancer by site and gender in Finland. The annual age-standardized incidence of laryngeal cancer is 3.2 per 100,000 for men and 0.3 per 100,000 for women. The proportion of females in the five university centers reviewed was 5%. During the 30-year time period from the early 1960s to the 1990s there has been a significant decrease in the laryngeal cancer incidence rates for males, but no change for females. Finland thus seems to be the only Western country with clearly declining occurrence rates in recent decades. In the same time period a clear decrease in the incidence of bronchial cancer has occurred in males and an increase in females. The results also show that in all five university hospital districts glottic cancer became more common (50-68%) in the late 1980s and 1990s, which is opposite to the situation in the 1960s when supraglottic localizations (65%) dominated in Finland. PMID- 10456275 TI - Free-flap reconstruction for laryngeal preservation after partial laryngectomy in patients with extended tumors of the oropharynx and hypopharynx. AB - Partial laryngeal resection often results in major aspiration problems, making larynx preservation during surgical removal of tumors of the oropharynx and hypopharynx impossible. However, free flaps can be used to reconstruct perilaryngeal tissue, thus preserving the larynx and ensuring a better quality of life for patients. We present the results of forearm free-flap reconstruction of the supraglottis in 22 patients who underwent resections of extended squamous cell carcinomas of the oropharynx and hypopharynx. A total of 9 patients had T3 lesions and 13 had T4 lesions. All patients were additionally treated with radiation therapy alone (to 70 Gy) or in combination with chemotherapy (Cisplatin; 5-fluorouracil). The mean follow-up was 2.4 years. In four patients, tracheostomy could be closed. Five patients suffered from severe aspiration, one of whom had to undergo a laryngectomy. Six patients had mild aspiration and 7 patients had no aspiration, but extensive edema made decanulation impossible. A total of 13 patients were free of disease, 4 patients died of disease, 1 patient died as the result of a second primary cancer and 1 patient died of other causes. Three patients are alive with persistent tumor. Although the majority of patients experienced a better quality of life as a result of larynx preservation, aspiration has remained a problem following treatment. PMID- 10456276 TI - Oncological and functional outcome of conservative surgery for primary supraglottic cancer. AB - The aim of this study was to verify the oncological and functional outcome of conservative surgical treatment of primary supraglottic squamous cell carcinoma (SGSCC) and related neck disease in order to verify the effectiveness of supraglottic laryngectomy (SL) and the validity of an "observation" policy in the control of clinically negative (N0) necks. Of a total of 252 consecutive patients affected by primary SGSCC seen between 1975 and 1990 at the Department of Otolaryngology of the University of Perugia (1975-1987) and the Catholic University of the Sacred Heart of Rome (1988-1990), a subset of 132 patients treated with classical SL was evaluated after presenting sufficient clinicopathological data and a follow-up period of at least 5 years. Tumors were staged according to the 1992 UICC TNM classification and grouped into stages I-II (n = 94) and III-IV (n = 38). Comprehensive neck dissections were performed only in the clinically positive (N+) necks (25/132 cases), while in the clinically N0 ones (107/132 cases) an "observation" policy under strict follow-up conditions was adopted. After primary surgery, the 5-year relapse-free survival (RFS) was 74%. The RFS was 80% for T1-2 disease and 65% for T3. The RFS was 80% for stages I-II tumors and 71% for stages III-IV. The actual 5-year overall survival (OS) was 89% for T1-T2 tumors and 67% for T3 disease or 93% for stage I-II and 69% for stages III-IV. The OS was 89% for N0 neck and 73% for N+. The 5-year-metastasis free survival (MFS) was 83% for N0 patients, 74% for N+, 84% for T1-T2 N0, 71% for T1-T2 N+, 81% for T3 N0 and 68% for T3 N+. In all, SL was found to be highly effective in the management of primary SGSCC. In the presence of clinically N0 neck "observation" under strict follow-up with therapeutic comprehensive neck dissection for delayed nodal recurrence, SL was suitable for controlling the neck cancer, as well as for salvaging recurrent disease. Bilateral elective, selective or functional neck dissection in every instance of supraglottic cancer was best performed only in those SGSCC patients who were more likely to have occult nodal disease on the basis of biological factors and imaging data. PMID- 10456277 TI - CO2 laser subtotal arytenoidectomy and posterior true and false cordotomy in the treatment of post-thyroidectomy bilateral laryngeal fixation in adduction. AB - A total of 39 patients with bilateral post-thyroidectomy vocal cord paralysis in adduction underwent CO2 laser subtotal arytenoidectomies with removal of the posterior third of the false and true vocal cords. Total airway resistance (Rtot) evaluated before and 4-10 months after surgery showed marked preoperative impairment before and significant improvement after surgery (P < 0.05). In five patients revision surgery was performed due to a progressive impairment of respiratory function. A variable degree of voice breathiness was observed after surgery; the maximum phonation time mean values were lower than normal and peak sound pressure levels 63 +/- 5 dB. In three cases aspiration was present in the first postoperative days, but swallowing dysfunctions disappeared within 1 week. Subtotal arytenoidectomy with removal of the posterior third of the true and false vocal folds was found to be a satisfactory surgical treatment for bilateral vocal cord paralysis in adduction. However, further research is still needed to define the surgical procedure able to balance respiratory, phonatory and sphincteric functions optimally. PMID- 10456278 TI - The Gregorio Maranon Hospital experience with vertical partial laryngectomies. AB - We present a retrospective study of 551 patients treated with conservative surgery for glottic carcinoma at the Gregorio Maranon Hospital between 1962 and 1996. In all, 12% of cases were locally advanced carcinomas. In early-stage carcinomas there were no statistical differences in 5-year survival between those treated by endoscopic laser resection, vertical hemilaryngectomy and radiotherapy. However, tumor recurrence after primary radiotherapy was higher (27%) than with conservative surgery (12%), while the voice preservation rate was significantly higher with surgery (83%) than with radiotherapy (72%). With locally advanced cancer, irradiated patients (to 60 Gy) had a 50% probability of recurrence with a very low chance for salvage by total laryngectomy (5-year survival rate, 38.5%). In contrast, partial laryngectomy could be performed on carefully selected patients, and the results for these patients were comparable to those for smaller lesions (with a 5-year survival rate of 81%). PMID- 10456279 TI - Expression of bcl-2 protein in lymphocytes of patients with laryngeal carcinoma. AB - The aim of this study was to evaluate the expression of bcl-2 protein in lymphocytes of the peripheral blood of patients with laryngeal carcinoma. The protein product of the proto-oncogene bcl-2 is a physiological inhibitor of apoptosis or programmed cell death. Since we believe that apoptosis is involved in the regulation of an immune response to a cancer process, we tried to show how this mechanism works in laryngeal carcinoma in comparison with normal peripheral blood lymphocytes of healthy controls. To explain the significance of this molecule's expression, we used flow cytometry to examine the expression of bcl-2 in T lymphocytes from the peripheral blood of 23 patients with laryngeal carcinoma and 20 healthy controls. Our study revealed that the expression of bcl 2 protein in T lymphocytes from the cancer patients was significantly higher than in the controls (P < 0.05). This difference in the expression of bcl-2 protein was found in both CD4 and CD8 subpopulations and was significantly higher than in the control group. In patients with laryngeal carcinoma expression of bcl-2 protein in T lymphocytes was higher in CD4 than in CD8 cells (P < 0.05). These results suggest that bcl-2 protein may interact in the regulation of apoptosis of lymphocytes, taking part in anti-cancer defence. PMID- 10456280 TI - Tumor angiogenesis in patients with laryngeal cancer. AB - Tumor angiogenesis was evaluated in 60 patients with primary laryngeal cancer by quantitating the microvessel density with antibodies against factor VIII. The microvessel density was then correlated with T stage, N stage, histologic grade and patient survival. A direct correlation was found between increased tumor angiogenesis and T stage, histologic grade and a shorter survival rate. PMID- 10456281 TI - A comprehensive analysis of selected diagnostic methods with respect to their usefulness in evaluating the biology of neoplastic cells in patients with laryngeal cancer. AB - The difficult and complicated mechanism of cancer development with little knowledge about the biology of existing cancers can lead to a permanent search for new examination techniques to improve the precision of life expectancy in patients and the selection of the most efficient methods of treatment. The aim of this study was to analyze certain prognostic factors, i.e., p53, Ki67, proliferating cell nuclear antigen (PCNA), DNA ploidy and cell proliferating activity, as well as the degree of morphological differentiation and cell maturity evaluated on an ultrastructural level in patients with laryngeal cancers in connection with data obtained from follow-up examinations and the clinical course of the disease. Neoplastic tissue was taken from 120 patients with laryngeal cancers. All underwent surgical treatment, radiotherapy and combined treatment in the Department of Otolaryngology. Karol Marcinkowski University School of Medical Sciences, Poznan, Poland, and the Department of Otolaryngology Head and Neck Surgery, Haukeland University, Bergen, Norway. Before beginning treatment all patients underwent histological verification of their neoplastic tissues. Histopathological examination proved that the majority of cases (95%) had a squamous cell carcinoma. The occurrence of changes within the lymph nodes of the neck (N) was significantly correlated with T, S, Ki67, metastases to lymph nodes, DNA ploidy, site and surgery performed. The degree of clinical progression (S) was intercorrelated with T, N, p53, Ki67, PCNA, DNA ploidy, site and laryngectomy. The occurrence of oncoprotein p53 in neoplastic cells was measured by the staining degree of their nuclei and was correlated with T, S, DNA ploidy, metastases to lymph nodes, PCNA and site. The degree of staining of neoplastic cells for the nuclear antigen Ki67 was correlated to T, N, G, S, DNA ploidy, metastases to lymph nodes and surgical treatment. The proliferative antigen PCNA in the examined population of patients was intercorrelated with T, p53, Ki67, metastases to lymph nodes and surgical treatment. The results obtained from DNA flow cytometry could be associated with N, G, p53, Ki67 and metastases to lymph nodes. On the basis of the results obtained, the techniques suggested for the morphological and biological evaluation of neoplastic cells in cancer of the larynx should include TNM classification + G + DNA + p53 + Ki67. PMID- 10456282 TI - Effects on voice by endolaryngeal microsurgery. AB - Endolaryngeal microsurgery (EM) is functionally oriented. Therefore, assessment of vocal function is important to evaluate the effect of the surgery on voice. In all, 58 patients, including 26 patients with vocal cord nodules and 32 patients with vocal cord polyps, underwent EM. The patients' voices were recorded and analyzed before EM and 2 weeks after. Analysis of voice quality included perceptual assessment and each patient's own subjective evaluation of social acceptability of voice according to the 10.0 cm visual analogue scale (VAS) scale. Acoustic voice signal data were measured for fundamental frequency (Fo), jitter, shimmer and normalized noise energy (NNE) using Tiger Electronics Dr. Speech software. Statistically significant (P < 0.001) improvement was achieved in both perceptual and acoustic analysis and in both patient groups. According to the VAS scale, a high degree of satisfaction with the surgery was achieved. The grade of hoarseness (G) as well as roughness (R) and breathiness (B) decreased significantly after the operation. EM resulted in a statistically significant decrease in the mean jitter, shimmer and NNE postoperatively. There were no significant changes in the Fo after EM. These results confirm a high degree of effectiveness of EM on vocal rehabilitation and meet the expectations regarding the assessment and documentation of postsurgical voice changes. PMID- 10456283 TI - Endoscopic high-frequency ultrasound of the larynx. AB - Over the last few years the development of new high-frequency ultrasound equipment has improved the application and resolution of endoluminal ultrasound. Following our in vitro study on the anatomical basis of endolaryngeal high frequency ultrasound we present the preliminary results of our sonographic examinations during microlaryngoscopy in order to characterize clinical applications. For the present study 38 patients underwent endolaryngeal ultrasound examinations. Laryngeal pathology included vocal fold polyps, laryngeal cysts, chronic laryngitis, Reinke's edema, epithelial dysplasia and cancer. The ultrasound examination was performed during microlaryngoscopy in a standardized pattern. A newly developed ultrasound apparatus was utilized with 10 MHz and 20 MHz catheters. Site, size and depth of the laryngeal lesion were evaluated. In 23 examinations of laryngeal carcinomas tumor size and infiltration could be measured and involvement of the thyroid cartilage or anterior commissure could be visualized. The exact extension of laryngeal cysts could also be seen. Ultrasound added no additional information to the endoscopic impression of the other laryngeal lesions. Present findings suggest that endoluminal high-frequency ultrasound might supplement microlaryngoscopy in the assessment of certain laryngeal lesions, especially for the evaluation of the size and infiltration of laryngeal tumors and to perform a more accurate preoperative staging without using computed tomography or magnetic resonance imaging. PMID- 10456284 TI - Ethics and clinical trials. AB - The current reference guideline about ethics in clinical trials is the Declaration of Helsinki of human rights in medical research. Three major principles are emphasised: respect of the patient to accept or not to participate in a trial, the constraints and the presumed risks must be acceptable for patients included in a study, and vulnerable subjects should not participate in studies. The investigator is responsible for obtaining a free and well-informed consent from patients before their inclusion in a study. Where possible, a new drug should always first be compared to placebo in order to prove its superiority. Else, a small-sized trial comparing a new drug versus a reference treatment can lead to an erroneous conclusion of absence of difference. Moreover, good results or improvement are obtained in at least 30% of cases with placebo, whatever the disease. The use of placebo is unethical in life-threatening diseases and when an effective proved drug exists. The use of placebo is ethical in severe diseases with no efficient drug, in some severe diseases even when an active reference treatment is available, and in all moderate and functional diseases. In order to detect flawed studies, most journals now ask for any manuscript submitted and reporting results of a randomised clinical trial to join a checklist in order to verify the quality of the trial. Finally, it remains the responsibility of the doctor to decide whether or not a protocol is ethical, to participate or not and to include patients or not. PMID- 10456285 TI - Reduction in QT dispersion and ventricular arrhythmias by ischaemic preconditioning in anaesthetized, normotensive and spontaneously hypertensive rats. AB - QT dispersion is a marker for dispersion of ventricular repolarization and electrical instability of the heart. However, QT dispersion remains undocumented in both normotensive rats (NTRs) and spontaneously hypertensive rats (SHRs), in particular in conditions of myocardial ischaemia/reperfusion (isch./rep.) and ischaemic preconditioning (IP). Therefore, we assessed the effects of IP on the dynamic change of QT and QTc dispersion during isch./rep., and on isch.- and rep. induced ventricular arrhythmias in both NTRs and SHRs. Isch. and rep. were produced by occlusion and release of a snare around the left coronary artery in all rats. The effect of IP (three cycles of 3 min coronary artery occlusion and 5 min rep.) on myocardial repolarization and on development of isch.- and rep. induced ventricular arrhythmias was studied in 12 NTRs and 12 SHRs. Another 12 NTRs or 12 SHRs were subjected to 10 min of isch. followed by 10 min rep. without IP. SHRs have significantly longer QT- and QTc-intervals as well as QT and QTc dispersion before isch. compared to NTRs. Myocardial isch. and early rep. largely increased QT and QTc dispersion in both NTRs and SHRs and resulted in a high incidence of isch.- and rep.-induced ventricular tachycardia (VT) and fibrillation (VF). IP significantly reduced QT and QTc dispersion in SHRs before isch., and remarkably reduced the elevation of QT and QTc dispersion during a prolonged period of isch. and rep. in all rats. This protective effect on electrophysiology of IP was associated with an antiarrhythmic effect against both isch.- and rep.-induced ventricular arrhythmias in NTRs and SHRs. Our data indicate that: 1) SHRs have a significantly higher baseline dispersion of ventricular repolarization than NTRs; 2) IP provides protection against ventricular arrhythmias in SHRs; 3) the increasing QT dispersion provoked by myocardial isch. and rep. is associated with a high incidence of isch.- and rep. induced ventricular arrhythmias and; 4) the reduction of QT dispersion by IP may be involved in its protective effect against isch.- and rep.-induced arrhythmias in both NTRs and SHRs. PMID- 10456286 TI - Histamine H3-receptor stimulation is unable to modulate noradrenaline release by the isolated rat heart during ischaemia-reperfusion. AB - The aim of this study was to evaluate the ability of H3-histaminergic prejunctional receptors to modulate the noradrenaline release induced by myocardial ischaemia in the rat, and the effects of an eventual modulation on haemodynamic, biochemical and electrophysiological parameters. Isolated rat hearts were perfused according to the Langendorff technique. Control hearts (n = 13) were not treated; two groups were treated with the H3-agonist R-alpha-methyl histamine at 0.3 microM (n = 14) and 1 microM (n = 11) and one group, used as positive control, was treated with the selective alpha 2-agonist Mivazerol at 0.5 microM (n = 14) added to the perfusion medium. Noradrenaline, lactate and transaminase output in the coronary effluent, as well as various haemodynamic and electrophysiological parameters, were measured during global and total ischaemia (30 min) and reperfusion (30 min). alpha 2-receptor stimulation increased ischaemia-induced noradrenaline release during reperfusion (195 +/- 13 vs. 145 +/ 12 pmol.g-1 in control group, P < 0.05). In contrast, R-alpha-methyl-histamine, at both doses, did not significantly modify these parameters. Both treatments did not affect ischaemia- and reperfusion-induced haemodynamic (decrease in heart rate or in left ventricular developed pressure), biochemical (lactate and GOT release) and electrophysiological (arrhythmias or increase in action potential duration) alterations. Unlike other species, the rat appears to be insensitive to H3-histaminergic receptor modulation of ischaemia-induced noradrenaline release, although a modulation can be seen with other prejunctional receptor agonists. PMID- 10456287 TI - Endothelin, but not angiotensin II, contributes to the hypoxic contractile response of large isolated pulmonary arteries in the rat. AB - The aims of this study were to investigate whether angiotensin II and/or endothelin could contribute to the hypoxic contractile response of isolated rat pulmonary artery. Experiments were performed for 1 h on noradrenaline precontracted arterial rings in hypoxic conditions (95% N2 and 5% CO2). Nicardipine, lisinopril, losartan, phosphoramidon, FR139317 and bosentan were used to block Ca2+ channels, angiotensin I-converting enzyme, AT1 receptors, endothelin-converting enzyme, ETA receptors, and ETA/ETB receptors, respectively. The profile of the hypoxic contractile response was biphasic, displaying, after a short relaxation, a weak and transient contraction (from 2-4 min) and then, before complete relaxation, a slowly developed but sustained contraction (from 14 60 min). Endothelium removal abolished the transient contraction and reduced ( 59%) the sustained contraction. Nicardipine did not modify the transient contraction, but concentration-dependently decreased (from -35% to -100%) the sustained contraction (P = 0.024). Lisinopril and losartan did not affect the response (P = 0.418 and P = 0.973, respectively). Bosentan did not modify the transient contraction, but concentration-dependently decreased (from -14% to 71%) the sustained contraction (P = 0.016), whereas phosphoramidon and FR139317 did not affect the response (P = 0.830 and P = 0.806, respectively). CONCLUSIONS: In rat, (i) both phases of the hypoxic contractile response are endothelium dependent and independent of angiotensin II; (ii) the transient contraction does not depend on endothelin; (iii) the sustained contraction, which involves calcium influx, appears partly dependent on mature endothelin released from storage granules by stimulating ETB receptors. PMID- 10456289 TI - Post-stroke treatment with imidapril reduces learning deficits with less formation of brain oedema in a stroke-prone substrain of spontaneously hypertensive rats. AB - The present study was undertaken to examine the effects of the ACE (angiotensin converting enzyme) inhibitor imidapril, on the brain, when administered after the onset of stroke in a stroke-prone substrain of spontaneously hypertensive rats (SHRSP). Learning deficits and induced lesions in the brain as well as in the kidneys and heart were investigated in detail. SHRSP were divided into two groups with or without salt loading at the age of 4 weeks. The salt loading was performed for 7-9 weeks to increase the incidence of stroke. Within 24 h after the first observation of stroke, animals were subsequently treated with 5 mg/kg imidapril orally once a day or the vehicle for up to the age of 27 weeks. Imidapril attenuated progression of neurological abnormalities such as irritability, hyperkinesia and motor dysfunction, and increased survival rate. In three-panel runway testing, learning deficits did not develop significantly in the imidapril-treated group, and was comparable to that in the non-salt loaded/non-stroke group. Imidapril reduced oedema formation in the cortex, hippocampus and striatum, and also suppressed lesion formation in the kidneys and heart. Imidapril thus suppressed progression of neurological deficits with loss of learning ability following onset of stroke, and also suppressed formation of oedema in the brain and decreased the number of lesions in other organs. Imidapril-induced reduction of cerebrovascular damage, which presumably occurs in the brain after stroke, may account for the inhibitory effects of imidapril on lesion formation and learning impairment. PMID- 10456288 TI - Soman-induced hypertension in conscious rats is mediated by prolonged central muscarinic stimulation. AB - The acetylcholinesterase inhibitor, soman, induces marked and sustained hypertension and tachycardia associated with a convulsive syndrome in rats. The aims of the present study were to distinguish between the cardiovascular and convulsant effects of soman and to determine whether the maintenance of the soman induced hypertension and tachycardia depends solely on a central muscarinic effect. To this end, using a computerised analysis of blood pressure (BP) in conscious freely moving rats, we examined the consequences on the increase in mean BP (MBP) and heart rate (HR) induced by soman (60 micrograms/kg, i.v.) of 1) a pre-treatment with the anticonvulsant drug diazepam (3 mg/kg, i.v.) and 2) atropine sulphate (10 mg/kg, i.v.) administered 10 or 60 min after the intoxication. Pretreatment with diazepam prevented the convulsions, assessed by electroencephalogram (EEG) recording, but modified neither the magnitude nor the kinetics of the pressor and tachycardic effects of soman (delta MBP = 74 +/- 2 and 73 +/- 5 mmHg, delta HR = 69 +/- 10 and 79 +/- 7 bpm, maximum MBP = 186 +/- 3 and 182 +/- 6 mmHg, maximum HR = 545 +/- 9 and 522 +/- 16 bpm in solvent- (n = 8) and diazepam- (n = 8) pre-treated rats, respectively). Whatever its time of administration, atropine sulphate fully and immediately reversed the rise in BP induced by soman. The soman-induced tachycardia was also suppressed by atropine administered 10 min after soman whereas it persisted when atropine was injected 60 min after the intoxication. These results show that the cardiovascular effects of soman can occur independently of the convulsive syndrome and that the maintenance of the soman-induced hypertension depends entirely on a permanent central muscarinic stimulation. PMID- 10456290 TI - Functional, endogenously expressed corticotropin-releasing factor receptor type 1 (CRF1) and CRF1 receptor mRNA expression in human neuroblastoma SH-SY5Y cells. AB - Corticotropin-releasing factor (CRF) is a hypothalamic 41-amino acid peptide which stimulates corticotropin (ACTH) release from the anterior pituitary and is also involved in the body response to stress. CRF1 receptors represent a potential target for novel antidepressant/anxiolytic drugs. The aim of the present study was to search for a human cell line expressing native, functional CRF1 receptors as a starting material for screening purposes. We identified CRF1 receptors functionally coupled to cAMP formation in human neuroblastoma SH-SY5Y cells. CRF induced concentration-dependent increases in cAMP accumulation in SH SY5Y cells (maximal increase 6.9 +/- 0.9 fold over basal values, n = 14). This effect was mimicked by related peptides with similar potencies: (mean pEC50 value) human/rat CRF (8.63), rat urocortin (9.32), sauvagine (8.97), urotensin I (8.93), ovine CRF (8.81). The efficacies of these agonists were nearly the same, with the exception of ovine CRF which was slightly less efficacious (75% the Emax of CRF). The responses to CRF were competitively antagonised by the following peptide fragments (mean pKB value): alpha-helical-CRF (9-41) (7.54), [D Phe12,Nle21,38,C alpha MeLeu37]CRF (12-41) (8.36) and [D-Tyr12]astressin (9.49) and by the selective, non-peptidic CRF1 receptor antagonists, CP-154,526 (7.76) and antalarmin (9.19). Estimation of receptor density by [125I]Tyr0-ovine CRF saturation binding yielded a modest number of binding sites (Bmax 12 fmol/mg protein, KD 0.2 nM). Analysis of mRNA by reverse transcription-polymerase chain reaction clearly revealed the presence of mRNA for CRF1 receptors in SH-SY5Y cells. A slight signal for CRF2 receptor mRNA was also observed. We conclude that neuroblastoma SH-SY5Y cells are endowed with native CRF1 receptors positively coupled to cAMP formation. They therefore constitute a useful functional model for the search of CRF1 selective compounds with potential anxiolytic/antidepressant activity. PMID- 10456291 TI - Effect of povidone-iodine on wound healing in control, diabetic and steroid depressed rats. AB - A comparative study was made on the effect of povidone-iodine on wound healing in normal, diabetic and steroid depressed states in the excision wound model in rats. Healing was assessed by the rate of contraction of wounds and epithelialization after three weeks of topical application. Normal and diabetic groups were comparable (P < 0.02) concerning the above-mentioned parameters as well as collagen formation. The steroid group showed significant retardation in healing time (P < 0.001), epithelialization (P < 0.001) and collagen formation (P < 0.001) showing that povidone-iodine did not overcome the steroid effect. PMID- 10456292 TI - The roles of P-glycoprotein and intracellular metabolism in the intestinal absorption of methadone: in vitro studies using the rat everted intestinal sac. AB - Methadone is used as a treatment for opiate detoxification in methadone maintenance programs. Intra- and inter-patient variations in methadone bioavailability have been observed after oral methadone treatment and this makes it difficult to predict a dosing regimen. Intestinal absorption and metabolism could explain these variations. The in vitro gut sac model was used to study the intestinal absorption of methadone, and it confirmed that methadone is a substrate for P-glycoprotein. The transport of methadone was increased in presence of P-gp inhibitors verapamil and quinidine. The appearance of a major metabolite of methadone, 2-ethylidene-1, 5-dimethyl-3, 3-diphenyl pyrrolidine (EDDP) in the gut sac contents also demonstrated the existence of intestinal metabolism of methadone. PMID- 10456293 TI - Effects of cigarette smoking on short-term variability of blood pressure in smoking and non smoking healthy volunteers. AB - The present trial was planned to study the effects of smoking on short-term variability of blood pressure and on haemodynamic parameters after an overnight cessation and after one day of repeated smoking in healthy cigarette smoking volunteers, compared to a control group of non-smokers who were not asked to smoke. 40 healthy male volunteers, 20 smokers and 20 non-smokers, participated in an open study with two period of measurements over a single day (morning and afternoon). Blood pressure and heart rate were measured using standard and finger recordings over 6 min before and 10 min after smoking one cigarette (in smokers only). During the two periods, smokers were asked to smoke 4 cm of a cigarette containing 1 mg of nicotine in 2 min, and a blood sample was taken for a plasma nicotine assay. In the smoking group, smoking the first cigarette of the day caused a significant increase of systolic blood pressure (+7%), diastolic blood pressure (+10%) and heart rate (+25%). The blood pressure variability in the frequency range of the Mayer waves (66-129 mHz) was increased after an overnight cessation of smoking in the smoking group in comparison to the non-smokers, and decreased significantly after the first cigarette of the day (7.1 +/- 4.0 to 3.2 +/- 1.8 mmHg2; P < 0.01). The changes observed in the afternoon after continuous smoking were significantly less important (3.8 +/- 1.9 to 3.2 +/- 1.9 mmHg2; NS). In the non-smoking group, the different parameters remained stable between the different measurements. These results suggest that an overnight cessation of smoking in smoking subjects is associated with a increase in sympathetic activity to the vascular system in the morning, which is released by smoking the first cigarette. This effect of smoking is reduced in the afternoon after a continuous nicotinic impregnation. PMID- 10456294 TI - Intra-individual variability and influence of urine collection period on dextromethorphan metabolic ratios in healthy subjects. AB - The aim of the study was to evaluate intra-individual variability in metabolic ratios (MRs) of dextromethorphan (DM) in healthy volunteers and to compare the MRs in urine collected 0-4, 0-8 and 0-24 h post-dose. Urinary molar ratios of DM to dextrorphan (MR1) and of DM to methoxymorphinan (MR2) were obtained after a single oral 27.5 mg dose of DM hydrobromide to ten healthy male and four female Caucasians (ten extensive metabolizers (EM) and four poor metabolizers (PM) of DM) to probe activities of CYP2D6 and CYP3A, respectively. Seven EM and one PM received DM on three additional occasions within 2 months. For the seven EM, the intra-individual variability (CVw) in the MRs obtained in the three urine collections ranged from 11 to 93% (MR1) and from 8 to 77% (MR2). The mean CVw estimated separately for the 4, 8 and 24 h urines by two-way analysis of variance reached 58, 57 and 44% for the MR1 and 50, 42 and 31% for the MR2, respectively. For all 14 subjects, the log-transformed ratios (MR1) obtained in the 24 h urines were highly correlated with those in either the 8 h (rs = 0.967, P < 0.0001) or 4 h urines (rs = 0.946, P < 0.0001). Correlation between the log-transformed MR2s were weaker (24 h vs. 8 h: rs = 0.829, P < 0.0001, 24 h vs. 4 h: rs = 0.831, P < 0.0001). The MR1s in 4 h and 8 h urines were only 2 and 9% less than those in 24 h urines (median differences) and varied from 48 and 47% below to 85 and 55% above (95% -CI for the differences). However, the MR2s in the 4 h and 8 h urines were shifted towards higher values by 49 and 23% and the corresponding 95% -CI limits were: 16-164% (4 h vs. 24 h) and 30-119% (8 h vs. 24 h). In conclusion, MR1 values in the 4 h urine collection agree well with those in longer collections and their use in epidemiological studies can be recommended. The intra-individual variability of approximately 50% in the MR1 has to be taken into account in clinical studies with within-subject design. Accurate determination of the MR2 requires at least a 24 h period of urine collection. PMID- 10456295 TI - Year 2000 contingency planning. AB - The clock is ticking. The year 2000 is just a few short months away, and hospitals have very little time to finish their Y2K preparations. With this in mind, we present this article--derived in part from ECRI's Special Report, Five Minutes to Midnight: Practical Y2K Contingency Plans for Healthcare Facilities- to help hospitals develop, refine, and implement sound, practicable contingency plans given the existing time constraints. Unlike the Special Report, which addresses all aspects of developing and implementing facilitywide contingency plans, this article focuses on departmental contingency planning needs related specifically to medical devices and systems. Department managers and others responsible for parts of the contingency planning process can use this article as a resource when reviewing and refining aspects of their plans, as well as when educating staff about the contingency planning process and about what will be expected of them in the coming months. In addition, although planning efforts should by now be well under way, departments that are still in the preliminary stages of developing a plan will find useful information to help them prepare for the year 2000. PMID- 10456296 TI - Y2K compliance report. Y2K hazard: radiotherapy systems could cause patients to be exposed to inappropriate radiation levels. PMID- 10456297 TI - Unexpected exposures possible with GE AMX-4 mobile x-ray units. PMID- 10456299 TI - Hazard report. Lethal risk: valve failure in Siemens 900 series ventilators can stop ventilation. PMID- 10456298 TI - Talk to the specialist. Pre-use testing of manual resuscitators. PMID- 10456300 TI - User experience network. Cable failure causes frequent alarms--and loss of control--on Hill-Rom Air-Shields C2HS Isolette infant incubators. PMID- 10456301 TI - [Safe motherhood: is the initiative an orphan?]. AB - Ten years after the Safe Motherhood Initiative has been set up, maternal morbidity and mortality remain a major public health issue in most developing countries. This presentation reviews the background of the initiative and the strategies adopted in Nairobi. Then, it undertakes an analysis of recent developments in maternal health policies. The authors discuss the comparative impact on maternal mortality of efforts aimed at a global improvement of the socio-economic and educative status of women, family planning activities, traditional birth attendants training programmes, and antenatal care. They stress the importance of the effective accessibility to life saving obstetric. Finally, they advocate a major involvement by gynecologist-obstetricians in the development and in the implementation of safe motherhood programmes. PMID- 10456302 TI - [Is systematic clinical breast examination in women still indicated?]. AB - Screening for breast cancer is not generalized in France. In order to evaluate any change in clinical practice, we reviewed three cohorts of one hundred successive breast cancers diagnosed in a geographical area without a breast screening campaign, starting in 1978, 1999 and 1996. The proportion of T1 (UICC) shifted from 32% to 50% and the rate of positive nodes among these T1 patients from 63% to 26% from the 1978 cohort to the 1996 cohort. There was an unchanged proportion (20%) of T4 patients in all three cohorts. Conservative treatment improved from 32% to 78% (p > 0.001). Systematic clinical breast examination should be strongly encouraged to lower the high proportion of T4 patients. PMID- 10456303 TI - [Tolerance of synthetic tissues and vaginal surgery. Report of 287 cases]. AB - OBJECTIVE: In this presentation, we attract attention to the disadvantage of using synthetic material in functional surgery of prolapse and urinary incontinence. PATIENTS AND METHODS: Two hundred eighty seven vaginal operations using synthetic material (Gore Tex, Dacron and Lyodura) were followed. The operations were: Mouchel procedure (127 cases), small slings (118 cases), large slings (11 cases), Stamey procedure (8 cases) and patch for paravaginal repair (23 cases). These operations were performed from 01/01/89 to 31/12/95. RESULTS: Mean follow-up at study end was 49 months. The intolerance phenomenon, leading to rejection, occurred between 1 and 72 months. The Mouchel procedure gave a rejection rate of 28.3% versus 9.3% for the slings (p < 0.001). Dacron was better tolerated (rejection rate at 19.3%) than Gore tex (rejection rate at 30.2%), p < 0.05. CONCLUSION: The substratum of the intolerance process would have two explanations (infection and foreign body reaction) for the early and late rejections. We suggest that the synthetic tissue tolerance is proportional to the exposed surface and to the distance which separates it from the vaginal scar. The ideal synthetic mesh material for pelvic surgery has yet to be developed. PMID- 10456304 TI - [Clinical aspects of primary cancer of the fallopian tube. A retrospective study of 20 cases]. AB - OBJECTIVES: From a review of 20 cases of primary carcinoma of the fallopian tube, we stress the various clinical aspects of the disease and discuss the possibilities of a preoperative diagnosis. MATERIALS AND METHODS: A retrospective study covering the period between 1982 and 1996. An analysis of the different etiological factors mentioned in the literature, of the physical signs and symptoms of the carcinoma, of the patients' age and hormonal status as well as the role of additional investigations was conducted. RESULTS: The average age of the patients was 62.3 years, 90% of the women had reached menopause and 15% were nullipara. A history of salpingitis was noted in 5% of the patients and another 5% had infertility problems. 20% of the patients had suffered from another carcinoma. Metrorrhagia was the most frequent clinical finding revealing the disease (55%) followed by abnormal secretions of blood stained liquid (15%). Hydrops Tubae Profluens was not observed in this series. The main complementary investigations performed were pelvic ultrasound scanning and hysterosalpingography, preoperative diagnosis was possible in 15% of cases; diagnosis was made during the operative procedure in 45% of cases and by pathologic examination in the remaining 40% of patients. CONCLUSION: Primary carcinoma of the fallopian tube is a rare form of cancer of unknown aetiology. Clinical signs are often unrelated and sometimes misleading; however, preoperative diagnosis is possible if one keeps in mind the existence of this carcinoma. PMID- 10456305 TI - [Change in sexual desire in the menopausal woman: a succinct evaluation]. AB - Physicians or gynecologists, specifically in their general practice or in the setting of a menopause clinic, are more and more frequently confronted with sexual complaints of menopausal women. Among these, decline in sexual desire is probably the most usually reported. The first study to evaluate a potential relationship between sexual functioning and menopause was conducted by Hallstrom in 1977. Thereafter, a review of the literature was able to show that there is nearly a consensus regarding the role of estrogens in that condition. They effectively relieve vaginal atrophy and resulting dyspareunia. There is less agreement, however, regarding a direct effect of estrogens on more complex sexual behavior and motivation. When analyzing potential influence of sex hormones, estrogens may exert a positive effect on the quality of the sexual relationship whereas androgens can definitely increase sexual "motivation" including sexual desire. In spite of the potentially important part played by androgens as promoters of libido and in the maintenance of sexual functioning in men and women, the exact role of the hormonal treatment in releaving sexual complaints still remains controversial. In some women where decline of sexual desire can be reasonably attributed to menopause, androgens in non-masculinizing adequate dosages, can be effectively included in the postmenopausal hormone replacement regimen. However, etiology of diminished sexual motivation and desire is far from univocal particularly in the human being where psychological, social and cultural influences are endowed with a prominent importance. It is accordingly quite conspicuous that our sexual life is not reduced to hormonal fluctuations only. A short critical review of the literature devoted to the main aspects of changes of sexual desire associated with menopause is presented. PMID- 10456306 TI - [Ultrasonography of placenta previa at the third trimester of pregnancy: research for signs of placenta accreta/percreta and vasa previa. Prospective color and pulsed Doppler ultrasonography study of 45 cases]. AB - OBJECTIVES: In a population of 45 placenta previa observed at third trimester of gestation we have tried to make the prenatal diagnosis of placenta percreta and vasa previa with color and pulsed Doppler ultrasonography. MATERIALS AND METHODS: We used a transabdominal sonography with full bladder and a transvaginal sonography with empty bladder. The first images obtained showed 20 placenta previa over the internal cervical os, 3 marginal and 22 low-lying placenta previa. We tried to find evidence of placenta percreta with gray-scale ultrasonography (loss of normal hypoechoic retroplacental myometrial zone, focal disruption of the uterine serosa and surrounding tissues, presence of intra placental lacunae) and with color and pulsed Doppler (arterial vessels with a diastolic flow value less than the flow value of a spiral artery behind the placenta, arterial vessels crossing from the placenta to surrounding tissues, intraplacental lacunae with arterial flow). We tried to find evidence of vasa previa in color and pulsed Doppler (a fetal vessel in seen above the lower segment of the uterus and below the fetal head. There is no change in the location of the vessel despite positional changes in both mother and fetus). The positivity of one sign in gray-scale ultrasonography or in color and pulsed Doppler led us to believe that the patient was affected by the anomaly. The final diagnosis of abnormal adherence of the placenta and of vasa previa was made on histological examination. RESULTS: Among the 20 placenta previa over the internal cervical os, we found 1 placenta percreta, 1 placenta accreta and 3 cases of vasa previa. In spite of our limited sample of cases of abnormal adherences, our results showed that gray-scale ultrasonography was sufficient to make a prenatal diagnosis of placenta accreta/percreta. Negative predictive value is 100% on a sample of 43 patients with no abnormal placental adherence. Color and pulsed Doppler brought no further evidence. In our population, color and pulsed Doppler had 100% positive predictive value for diagnosis of vasa previa. CONCLUSION: We found the only 2 cases of abnormal adherent placental fragments and the 3 cases of vasa previa present in our population. The study of the lower segment of the uterus should be thorough as abnormal zones may be small-sized. Color Doppler is the reference technique for sighting vasa previa and gray-scale ultrasonography for abnormal adherences of the placenta. PMID- 10456307 TI - [Medical abortion in the second and third trimester. Report of 125 indications from 1992 to 1995]. AB - OBJECTIVE: To verify the indications of second and third trimester termination of pregnancy in a fetal medicine department. METHODS: The medical files of 125 patients who underwent medically-induced abortion between 12 to 39 weeks over a four years period (1992-1995) in the French university hospital of Tours, were studied. The medical termination of pregnancy indications inventory allowed us to compare our study to those from the French literature. RESULTS: Sonographic diagnosis was the main departure point for late abortion indications (70%). Fetal indications were predominant (43%), followed by chromosomal anomalies (30%). If obstetrical indications still existed, maternal indications were exceptional (3%). The reasons for termination were severe structural abnormalities which, in most cases, were not compatible with survival. All the decisions were taken by the physicians from the different specialties concerned by the pathology with respect of the parents opinion. CONCLUSION: With generalization of fetal medicine departments with consulting committees for prenatal diagnosis, misguided abortions can be avoided; but an accurate law, standardized in all European countries, would be necessary. PMID- 10456308 TI - [Scarred uterus: is routine exploration of the cesarean scar after vaginal birth always necessary?]. AB - OBJECTIVE OF THE STUDY: To determine the risks and benefits of routine transcervical revision of previous cesarean uterine scar in patients with successful vaginal delivery. MATERIAL AND METHODS: Retrospective study, over a 10 year period, of a routine palpation practice in the two units of our Obstetric department. Then, a 30-month prospective study comparing, in each unit, two different attitudes toward uterine revision (routine exploration vs symptomatic patients exploration) was conducted. RESULTS: The retrospective part of our study led us to report 3 uterine ruptures (0.43% of all scarred uterus) and 14 dehiscences (2%) during the ten years. All uterine ruptures were sufficiently symptomatic in order to be suspected prior to scar exploration. No dehiscence needed surgical treatment. Some patients with bloodless dehiscence and no repair had subsequent vaginal deliveries with no scar separation found on uterine exploration. In the prospective part of our study, we found a significative difference in the occurrence of fever (18.9% vs 9.9%; p < 0.05) and antibiotic treatment (22.8% vs 12.7%; p < 0.05) between the two groups based on attitude toward uterine revision. CONCLUSION: These data suggest that transcervical revision of previous cesarean uterine scar should be performed only in symptomatic patients (persistent suprapubic pain, placental retention, excessive bleeding during labor or delivery) or when risk factors are present (prolonged labor, prolonged expulsive efforts, instrumental extraction). PMID- 10456309 TI - [Labor in the squatting position. [A randomized trial comparing the squatting position with the classical position for the expulsion phase]. AB - OBJECTIVE OF STUDY: To evaluate the influence of a vertical delivery position squatting during expulsion, studying different parameters such as duration of expulsion, neonatal status, delivery mode, frequency of hemorrhagia, perineum status and patient comfort. TYPE OF STUDY: Randomized, unicentric, comparative, open, prospective. MATERIAL AND METHOD: After a retrospective study determining feasibility, 240 patients were randomized to equal 2 groups in which the delivery was performed in squat position versus in lithotomy position. All the above described parameters were recorded and treated by the Epi-Info software package using Student t test, chi 2 and Kruskall-Wallis test. RESULTS: Our study has shown a tendency to shorten the length of the expulsion phase and a reduced use of forceps in the squat position. This is consistent with other studies published in the literature. We would like to emphasize that even if it does not show a medical advantage on the studied parameters, the squat position is not deleterious and can provide a better comfort for the patients who want to use it. PMID- 10456310 TI - [Parietal endometriosis in abdominal scars. Report of 3 cases]. AB - Parietal endometriosis is a rare disease. Its diagnosis and treatment are often difficult. We report 3 cases of parietal endometriosis occurring in cesarean and appendectomy scars. Clinical symptoms are not specific and may lead to erroneous diagnosis. Diagnosis is usually made on the histological exam of the resected lesion. Treatment of choice is complete surgical excision. PMID- 10456311 TI - [Acardiac acephalic fetus and twin pregnancy. Report of a case]. AB - The present case describes an acardiac, acephalic fetus, diagnosed by ultra sound, during the 33rd gestational week of an insulin dependent diabetic parturient, who had developed toxemia gravida. The diagnosis had been confirmed by the autopsy. The differential diagnosis from an in-utero death of the second twin had been raised. The acardiac acephalic foetus is a rare congenital malformation observed during the monozygotic twin pregnancy. The second fetus being normal, our approach consisted in the clinical observation of the parturient, followed by cesarean section delivery the 4th day of the 36th gestational week to avoid worsening of toxemia gravida. PMID- 10456312 TI - [Ethical recommendations 1998. Committee of the International Federation of Gynecology and Obstetrics (FIGO) on the Ethical Aspects of Human Reproduction and Women's Health]. PMID- 10456313 TI - Accommodation of foreign genes into the Sendai virus genome: sizes of inserted genes and viral replication. AB - Sendai virus (SeV) is an enveloped virus with a negative sense genome RNA of about 15.3 kb. We previously established a system to recover an infectious virus entirely from SeV cDNA and illustrated the feasibility of using SeV as a novel expression vector. Here, we have attempted to insert a series of foreign genes into SeV of different lengths to learn how far SeV can accommodate extra genes and how the length of inserted genes affects viral replication in cells cultured in vitro and in the natural host, mice. We show that a gene up to 3.2 kb can be inserted and efficiently expressed and that the replication speed as well as the final virus titers in cell culture are proportionally reduced as the inserted gene length increases. In vivo, such a size-dependent effect was not very clear but a remarkably attenuated replication and pathogenicity were generally seen. Our data further confirmed reinforcement of foreign gene expression in vitro from the V(-) version of SeV in which the accessory V gene had been knocked out. Based on these results, we discuss the utility of SeV vector in terms of both efficiency and safety. PMID- 10456314 TI - Structures of the O-specific polysaccharides and a serological cross-reactivity of the lipopolysaccharides of Proteus mirabilis O24 and O29. AB - Strains of Proteus mirabilis belonging to serogroups O24 and O29 are frequent in clinical specimens. Anti-P. mirabilis O24 serum cross-reacted with the lipopolysaccharide (LPS) of P. mirabilis O29 and vice versa. The structures of the O-specific polysaccharides (OPSs, O-antigens) of both LPSs were established using sugar analysis and one- and two-dimensional 1H- and 13C-NMR spectroscopy and found to be different. SDS-PAGE and Western immunoblotting suggested that the serological cross-reactivity of the LPSs is due to a common epitope(s) on the core-lipid A moiety, rather than on the OPS. Therefore, the epitope specificity and the structures of the O-antigens studied are unique among Proteus serogroups. PMID- 10456316 TI - Exciton levels structure of antenna bacteriochlorophyll c aggregates in the green bacterium Chloroflexus aurantiacus as probed by 1.8-293 K fluorescence spectroscopy. AB - We have demonstrated temperature-dependence of the steady-state fluorescence lineshape of the bacteriochlorophyll (BChl) c band measured for intact cells of the green bacterium Chloroflexus aurantiacus over the 1.8-293 K range. The measured temperature-dependence has been shown to be in good agreement with the theoretical one, calculated for our original model of pigment organization in the chlorosomal oligomeric antenna of green photosynthetic bacteria based on spectral hole-burning studies (Fetisova, Z.G. et al. (1996) Biophys. J. 71, 995-1010). This model implies that the BChl c antenna unit is a tubular aggregate of six exciton-coupled linear pigment chains having the exciton level structure with strongly allowed higher levels. PMID- 10456315 TI - Comparison of the effects of bax-expression in yeast under fermentative and respiratory conditions: investigation of the role of adenine nucleotides carrier and cytochrome c. AB - A new system for bax-expression in yeast has been devised to investigate bax's effect under fermentative and respiro-fermentative conditions. This has allowed us to show unambiguously that the ability of bax to kill yeast is higher under respiratory conditions than under purely fermentative conditions. The extent of killing under respiro-fermentative conditions (non-repressive sugars) is intermediate. It has been proposed that the two proteins adenine nucleotides carrier (ANC) and cytochrome c play a crucial role in bax-induced cell death. We have investigated the effects of deletion of the genes encoding the two proteins on the toxicity induced by bax, using this new system. The absence of ANC did not modify bax-induced lethality in any way. Moreover, the absence of cytochrome c also did not prevent bax-induced death. Only the kinetics of lethality were altered. All these effects are prevented by co-expression of bcl-xL. PMID- 10456317 TI - Identification of a novel gene of the X,K-ATPase beta-subunit family that is predominantly expressed in skeletal and heart muscles. AB - We have identified the fifth member of the mammalian X,K-ATPase beta-subunit gene family. The human and rat genes are largely expressed in skeletal muscle and at a lower level in heart. The deduced human and rat proteins designated as beta(muscle) (beta(m)) consist of 357 and 356 amino acid residues, respectively, and exhibit 89% identity. The sequence homology of beta(m) proteins with known Na,K- and H,K-ATPase beta-subunits are 30.5-39.4%. Unlike other beta-subunits, putative beta(m) proteins have large N-terminal cytoplasmic domains containing long Glu-rich sequences. The data obtained indicate the existence of hitherto unknown X,K-ATPase (most probably Na,K-ATPase) isozymes in muscle cells. PMID- 10456318 TI - Phosphorylation and activation of Ca2+/calmodulin-dependent protein kinase phosphatase by Ca2+/calmodulin-dependent protein kinase II. AB - Ca2+/calmodulin-dependent protein kinase phosphatase (CaMKPase) is a protein phosphatase which dephosphorylates autophosphorylated Ca2+/calmodulin-dependent protein kinase II (CaMKII) and deactivates the enzyme (Ishida, A., Kameshita, I. and Fujisawa, H. (1998) J. Biol. Chem. 273, 1904-1910). In this study, a phosphorylation-dephosphorylation relationship between CaMKII and CaMKPase was examined. CaMKPase was not significantly phosphorylated by CaMKII under the standard phosphorylation conditions but was phosphorylated in the presence of poly-L-lysine, which is a potent activator of CaMKPase. The maximal extent of the phosphorylation was about 1 mol of phosphate per mol of the enzyme and the phosphorylation resulted in an about 2-fold increase in the enzyme activity. Thus, the activity of CaMKPase appears to be regulated through phosphorylation by its target enzyme, CaMKII. PMID- 10456319 TI - Selection of ganglioside GM1-binding peptides by using a phage library. AB - Ganglioside Gal beta1 --> 3GalNAc beta1 --> 4(NeuAc alpha2 --> 3) Gal beta1 --> 4Glc beta1 -->1'Cer (GM1)-binding peptides were obtained from a phage-displayed pentadecapeptide library by an affinity selection. The selection processes were in situ-monitored by a quartz-crystal microbalance method, on which a ganglioside GM1 monolayer was transferred. After five rounds of biopanning, the DNA sequencing of 18 selected phages showed that only three individual clones were selected. The peptide sequences of the random region were found to be DFRRLPGAFWQLRQP, GWWYKGRARPVSAVA and VWRLLAPPFSNRLLP. Binding constants of these phage clones to the GM1 monolayer were 10(10) M(-1). Three synthetic pentadecapeptides inhibited the binding of cholera toxin B subunit to the GM1 monolayer with an IC50 of 24, 13 and 1.0 microM, respectively. These peptides will be useful for searching functional roles of ganglioside GMI. PMID- 10456320 TI - Flow cytometry and surface plasmon resonance analyses demonstrate that the monoclonal antibody JIM19 interacts with a rice cell surface component involved in abscisic acid signalling in protoplasts. AB - Abscisic acid (ABA) is a plant hormone involved in many developmental and physiological processes, but as yet, no ABA receptor has been identified. Flow cytometry of rice protoplasts and immunoblotting of purified plasma membranes (PMs) have been used to demonstrate that the monoclonal antibody JIM19 recognizes carbohydrate epitopes of cell surface glycoproteins. Using surface plasmon resonance technology specific binding of PMs to JIM19 was observed. Such interaction was antagonized significantly by ABA, but not by the biologically inactive ABA catabolite phaseic acid. These in vitro interactions were correlated with the biological activities of JIM19, ABA and phaseic acid on activation of the ABA-inducible Em promoter using two different transient reporter gene assays, beta-glucuronidase/luciferase and quantitative flow cytometry of Aequoria green fluorescent protein. Pre-treatment with JIM19 resulted in significant inhibition of ABA-inducible gene expression. Taken together, these data suggest that JIM19 interacts with a functional PM complex involved in ABA signalling. PMID- 10456321 TI - Synthesis of plasmalogens in eye lens epithelial cells. AB - The present paper describes cloning and sequencing of the mouse cDNA encoding dihydroxyacetonephosphate acyltransferase (DAPAT), the peroxisomal key enzyme of plasmalogen (PM) biosynthesis. Using monospecific antibodies, we localized DAPAT and alkyl dihydroxyacetonephosphate synthase to peroxisomes of mouse lens epithelial cells (LECs) and determined their enzymatic activity. By electrospray ionization mass spectrometry of mouse lens lipid extracts, we identified phosphatidyl ethanolamine including plasmenyl ethanolamine species as major constituents. Our data demonstrate the capacity of LECs to synthesize PMs and the high coincidence between deficiency of PM and early manifestation of cataract in patients with peroxisomal disorders suggests that ether-bonded lipids may play an important role in maintaining lens transparency. PMID- 10456322 TI - Molecular characterization of the 5' control region and of two lethal alleles affecting the hsp60 gene in Drosophila melanogaster. AB - The chaperonins are evolutionarily conserved essential cellular proteins that help folding newly synthesized or translocated proteins, spending ATP. We present here the molecular analysis of the hsp60 gene promoter region and of two Drosophila hsp60 ethyl methane sulfonate embryonic lethal alleles that have an identical phenotype. No heat shock element sequences were found in the 5' region, supporting previous data (Kozlova, T. et al., 1997) which suggests that mitochondrial Drosophila melanogaster HSP60.1 is not heat inducible. By sequencing the lethal allele's entire open reading frame (ORF), we found a C-T transition in the hsp60F409 allele that produces a serine to leucine change, apparently distorting the protein equatorial domain structure. No changes were found in the hsp60G93 ORF. However, an analysis of the heterogeneous nuclear RNA levels showed a reduction of the hsp60 transcript in hsp60G93 flies as compared to the wild-type. These data suggest that although the defects in the hsp60 gene produced by these alleles are at different levels, both behave as null mutations. PMID- 10456323 TI - hnRNPs H, H' and F behave differently with respect to posttranslational cleavage and subcellular localization. AB - hnRNPs H, H' and F belong to a subfamily of the hnRNPs sharing a high degree of sequence identity. Eukaryotic expression and specific C-terminal antibodies were used to demonstrate great variation in the intracellular fate of the proteins. hnRNPs H and H' become posttranslational cleaved into C-terminal 35 kDa proteins (H(C), H'(C)) and possibly into N-terminal 22 kDa proteins. No detectable cleavage was observed for hnRNP F. hnRNP H/H' is almost exclusively localized to the nucleus of many cell types while hnRNP F varies from a predominant nuclear localization in some cells to a predominant cytoplasmic localization in other cells. The different fates may reflect differences in functional roles that so far only have included nuclear functions. The presence of significant quantities of hnRNP F in the cytoplasm of many cells indicates that it also may have a functional role here. PMID- 10456324 TI - Low iron concentration and aconitase deficiency in a yeast frataxin homologue deficient strain. AB - Deletion of the yeast frataxin homologue, YFH1, elicits accumulation of iron in mitochondria and mitochondrial defects. We report here that in the presence of an iron chelator in the culture medium, the concentration of iron in mitochondria is the same in wild-type and YFH1 deletant strains. Under these conditions, the activity of the respiratory complexes is restored. However, the activity of the mitochondrial aconitase, a 4Fe-4S cluster-containing protein, remains low. The frataxin family bears homology to a bacterial protein family which confers resistance to tellurium, a metal closely related to sulfur. Yfh1p might control the synthesis of iron-sulfur clusters in mitochondria. PMID- 10456325 TI - Functional expression of the Na/K pump is controlled via a cyclosporin A sensitive signalling pathway in activated human lymphocytes. AB - An immunosuppressant cyclosporin A (CsA) inhibits T-cell proliferation by blocking the nuclear factor of activated T-cells (NFAT) required for expression of the interleukin-2 (IL-2) gene. This work has demonstrated for the first time that in human blood lymphocytes (HBLs) activated by phytohemagglutinin (PHA), CsA at anti-proliferative doses inhibits the late sustained increase in ouabain sensitive Rb(K) influxes, which accompanies the growth phase of G0/G1/S transition. CsA affects neither the initial, transient activation of the pump in response to PHA nor the ouabain-resistant ion fluxes during cell cycle progression. When the HBLs were rendered competent to proliferate by phorbol 12,13-dibutyrate ester and ionomycin in the presence of CsA, the exogenous IL-2 did not bypass the initial inhibitory effect of CsA on the long-term pump enhancement. When applied after the competence induction, CsA produced no effect on the sustained increase in ouabain-sensitive Rb influxes during the IL-2 induced progression phase. These results indicate that in activated HBLs, (1) IL 2 is involved in functional expression of the Na/K pump during cell transition from quiescence to proliferation, (2) the cell cycle-associated upregulation of the pump is related to a CsA-sensitive signalling pathway. PMID- 10456326 TI - Detection of a novel plasma serine protease during purification of vitamin K dependent coagulation factors. AB - A novel serine protease (PHBSP) was purified from human plasma by two chromatographic steps with a final yield of 1.6 mg/l plasma. The protease consists of two disulfide-bridged chains of about 50 and 30 kDa with the light chain containing the active site of the enzyme. NH2-terminal sequence analysis revealed identity to the deduced amino acid sequence of HGFA-like mRNA. The activity of PHBSP is strongly dependent on Ca2+ ions and is efficiently inhibited by alpha2-antiplasmin and aprotinin. Possible functions of PHBSP in the hemostatic system are discussed. PMID- 10456327 TI - The autocrine motility factor receptor gene encodes a novel type of seven transmembrane protein. AB - Autocrine motility factor receptor (AMFR) is a cell surface glycoprotein of molecular weight 78,000 (gp78), mediating cell motility signaling in vitro and metastasis in vivo. Here, we cloned the full-length cDNAs for both human and mouse AMFR genes. Both genes encode a protein of 643 amino acids containing a seven transmembrane domain, a RING-H2 motif and a leucine zipper motif and showed a 94.7% amino acid sequence identity to each other. Analysis of the amino acid sequence of AMFR with protein databases revealed no significant homology with all known seven transmembrane proteins, but a significant structural similarity to a hypothetical protein of Caenorhabditis elegans, F26E4.11. Thus, AMFR is a highly conserved gene which encodes a novel type of seven transmembrane protein. PMID- 10456328 TI - Computational modeling of the rate limiting step in low molecular weight protein tyrosine phosphatase. AB - Hydrolysis of the phosphoenzyme intermediate is the second and rate limiting step of the reaction catalyzed by the protein tyrosine phosphatases (PTPs). The cysteinyl phosphate thioester bond is cleaved by nucleophilic displacement where an active site water molecule attacks the phosphorus atom. Starting from the crystal structure of the low molecular weight PTP, we study the energetics of this reaction utilizing the empirical valence bond method in combination with molecular dynamics and free energy perturbation simulations. The reactions of the wild-type as well as the D129A and C17S mutants are modeled. For the D129A mutant, which lacks the general acid/base residue Asp-129, an alternative reaction mechanism is proposed. The calculated activation barriers are in all cases in good agreement with experimental reaction rates. The present results together with earlier computational and experimental work now provide a detailed picture of the complete reaction mechanism in many PTPs. The key role played by the structurally invariant signature motif in stabilizing a double negative charge is reflected by its control of the energetics of both transition states and the reaction intermediate. PMID- 10456329 TI - Temperature-dependent interaction of thermo-sensitive polymer-modified liposomes with CV1 cells. AB - Egg yolk phosphatidylcholine liposomes modified with a copolymer of N acryloylpyrrolidine and N-isopropylacrylamide having a lower critical solution temperature at ca. 40 degrees C were prepared and an effect of temperature on their interaction with CV1 cells was investigated. The unmodified liposomes were taken up by the cells approximately to the same extent after 3 h incubation at 37 and 42 degrees C. In contrast, uptake of the polymer-modified liposomes by CV1 cells decreased slightly at 37 degrees C but increased greatly at 42 degrees C, compared to the unmodified liposomes. Proliferation of the cells was partly prohibited by the incubation with the unmodified liposomes encapsulating methotrexate at 37 and 42 degrees C. The treatment with the polymer-modified liposomes containing methotrexate at 37 degrees C hardly effected the cell growth. However, the treatment at 42 degrees C inhibited the cell growth completely. It is considered that the highly hydrated polymer chains attached to the liposome surface suppressed the liposome-cell interaction below the lower critical solution temperature of the polymer but the dehydrated polymer chains enhanced the interaction above this temperature. Because interaction of the polymer-modified liposomes with cells can be controlled by the ambient temperature, these liposomes may have potential usefulness as efficient site specific drug delivery systems. PMID- 10456330 TI - Curcumin mediated apoptosis in AK-5 tumor cells involves the production of reactive oxygen intermediates. AB - Curcumin, the active ingredient of the rhizome of Curcuma longa has anti inflammatory, antioxidant and antiproliferative activities. Although its precise mode of action remains elusive, studies have shown that chemopreventive action of curcumin might be due to its ability to induce apoptosis in cancer cells. Curcumin was shown to be responsible for the inhibition of AK-5 tumor (a rat histiocytoma) growth by inducing apoptosis in AK-5 tumor cells via caspase activation. This study was designed to investigate the mechanism leading to the induction of apoptosis in AK-5 tumor cells. Curcumin treatment resulted in the hyperproduction of reactive oxygen species (ROS), loss of mitochondrial membrane potential (delta psi(m)) and cytochrome c release to the cytosol, with the concomitant exposure of phosphatidylserine (PS) residues on the cell surface. This study suggests redox signalling and caspase activation as the mechanisms responsible for the induction of curcumin mediated apoptosis in AK-5 tumor cells. PMID- 10456331 TI - Molecular cloning of haploid germ cell-specific tektin cDNA and analysis of the protein in mouse testis. AB - Tektins are a class of proteins that form filamentous polymers in the walls of ciliary and flagellar microtubules. We report here the molecular cloning of a new member of the tektin family, tektin-t, identified from a mouse haploid germ cell specific cDNA library. Tektin-t mRNA encodes a protein of 430 deduced amino acids possessing RSNVELCRD, the conserved sequence of tektin family proteins. Western blotting showed a single band having a molecular weight of 86 kDa in the mouse testis. Immunohistochemistry of the testis showed that tektin-t is localized in the flagella of elongating spermatids from developmental step 15 to maturity. PMID- 10456332 TI - Calmodulin inhibits inositol 1,4,5-trisphosphate-induced calcium release through the purified and reconstituted inositol 1,4,5-trisphosphate receptor type 1. AB - Our previous studies have demonstrated that calmodulin binds to IP3R type I (IP3R1) in a Ca2+ dependent manner, which suggests that calmodulin regulates the IP3R1 channel. In the present study, we investigated real-time kinetics of interactions between calmodulin and IP3R1 as well as effects of calmodulin on IP3 induced Ca2+ release by purified and reconstituted IP3R1. Kinetic analysis revealed that calmodulin binds to IP3R1 in a Ca2+ dependent manner and that both association and dissociation phase consist of two components with time constants of k(a) = 4.46 x 10(2) and > 10(4) M(-1) s(-1) k(d) = 1.44 x 10(-2) and 1.17 x 10(-1) s(-1). The apparent dissociation constant was calculated to be 27.3 microM. The IP3-induced Ca2+ release through the purified and reconstituted IP3R1 was inhibited by Ca2+/calmodulin, in a dose dependent manner. We interpret our findings to mean that calmodulin binds to IP3R1 in a Ca2+ dependent manner to exert inhibitory effect on IP3R channel activity. This event may be one of the mechanisms governing the negative feedback regulation of IP3-induced Ca2+ release by Ca2+. PMID- 10456333 TI - Enhanced transport of anticancer agents and leukotriene C4 by the human canalicular multispecific organic anion transporter (cMOAT/MRP2). AB - We established stable human canalicular multispecific organic anion transporter (cMOAT/MRP2) cDNA transfectants, CHO/cMOAT from non-polarized Chinese hamster ovary (CHO)-K1 and LLC/cMOAT from polarized pig kidney epithelial LLC-PK1. Human cMOAT was mainly localized in the plasma membrane of CHO/cMOAT and in the apical membrane of LLC/cMOAT. The ATP-dependent uptake of leukotriene C4 (LTC4) into CHO/cMOAT membrane vesicles was enhanced compared with empty vector transfectants. Km values in CHO/cMOAT membrane vesicles were 0.24 microM for LTC4 and 175 microM for ATP. Drug sensitivity to vincristine and cisplatin in human cMOAT cDNA transfectants decreased, but not to etoposide. Cellular accumulation of vincristine and cisplatin in human cMOAT cDNA transfectants decreased, but not of etoposide. The uptake of LTC4 into CHO/cMOAT membrane vesicles was inhibited by exogenous administration of vincristine or cisplatin, but not that of etoposide. Moreover, this inhibition was more enhanced in the presence of glutathione. These consequences indicate that drug resistance to vincristine or cisplatin appears to be modulated by human cMOAT through transport of the agents, possibly in direct or indirect association with glutathione. PMID- 10456334 TI - Glucokinase regulatory protein is essential for the proper subcellular localisation of liver glucokinase. AB - Glucokinase (GK), a key enzyme in the glucose homeostatic responses of the liver, changes its intracellular localisation depending on the metabolic status of the cell. Rat liver GK and Xenopus laevis GK, fused to the green fluorescent protein (GFP), concentrated in the nucleus of cultured rat hepatocytes at low glucose and translocated to the cytoplasm at high glucose. Three mutant forms of Xenopus GK with reduced affinity for GK regulatory protein (GKRP) did not concentrate in the hepatocyte nuclei, even at low glucose. In COS-1 and HeLa cells, a blue fluorescent protein (BFP)-tagged version of rat liver GK was only able to accumulate in the nucleus when it was co-expressed with GKRP-GFP. At low glucose, both proteins concentrated in the nuclear compartment and at high glucose, BFP-GK translocated to the cytosol while GKRP-GFP remained in the nucleus. These findings indicate that the presence of and binding to GKRP are necessary and sufficient for the proper intracellular localisation of GK and directly involve GKRP in the control of the GK subcellular distribution. PMID- 10456335 TI - Lack of heat shock response triggers programmed cell death in a rat histiocytic cell line. AB - Stress response is a universal phenomenon. However, a rat histiocytic cell line, BC-8, showed no heat shock response and failed to synthesize heat shock protein 70 (hsp70) upon heat shock at 42 degrees C for 30 min. BC-8 is a clone of AK-5, a rat macrophage tumor line that is adapted to grow in culture and has the same chromosome number and tumorigenic potential as AK-5. An increase in either the incubation temperature or time or both to BC-8 cells leads to loss of cell viability. In addition, heat shock conditions activated apoptotic cell death in these cells as observed by cell fragmentation, formation of nuclear comets, apoptotic bodies, DNA fragmentation and activation of ICE-like cysteine proteases. Results presented here demonstrate that BC-8 cells cannot mount a typical heat shock response unlike all other eukaryotic cells and that in the absence of induction of hsps upon stress, these cells undergo apoptosis at 42 degrees C. PMID- 10456336 TI - Solution conformation on bovine growth hormone releasing factor by 1H NMR and molecular modeling. AB - The structure of bovine growth hormone releasing factor (bGHRF) consisting of 44 amino acids has been studied in CD and 1H nuclear magnetic resonance (NMR) spectroscopy in conjunction with molecular modeling. Since bGHRF does not have an ordered structure in water alone, a 30% 2,2,2-trifluoroethanol (TFE) aqueous solvent was used to induce considerable alpha-helical structures, which corresponds to a helical content of approximately 62% as determined by circular dichroism (CD). The secondary structure was obtained from nuclear Overhauser enhancement and 3J(HN alpha) coupling constant in 30% TFE solution. Three dimensional structures consistent with NMR data were generated by using distance geometry calculation. A set of 267 interproton distances derived from nuclear Overhauser effect correlation spectroscopy (NOESY) experiments and coupling constants were used. From the initial random conformations, 50 distance geometry structures with minimal violations were selected for further refinement. The 14 best structures were obtained after simulated annealing calculation with energy minimization. The structure of bGHRF in 30% TFE solution was characterized by one alpha-helix (residues 8-19), two poorly constrained helices (residues 23-27 and residues 31-34) and a beta I(III)-turn fragment (residues 20-23; phi(i+1) = -53.1 degrees, psi(i+1) = -19.6 degrees, phi(i+2) = -59.9 degrees, psi(i+2) = -20.6 degrees) connected by the segments of less defined structures in N-terminal and omega-shaped flexible C-terminal determined from NOESY cross peaks between helical segment (residues 14-18) and tail fragment (residues 42-44). The obtained structure will play an important role toward the understanding of the structural and functional role of the GHRF. PMID- 10456337 TI - Treatment of granuloma faciale with the 585-nm pulsed dye laser. PMID- 10456338 TI - Management of cutaneous lymphomas using a multidisciplinary approach. PMID- 10456339 TI - Behavior of tissue-engineered skin: a comparison of a living skin equivalent, autograft, and occlusive dressing in human donor sites. AB - OBJECTIVE: To compare the behavior of a tissue-engineered living skin equivalent (LSE) with an autograft in acute donor site wounds. DESIGN: Paired-comparison, randomized control trial. SETTING: A university dermatology service. PATIENTS: Three donor sites were created on the anterior thigh of each of 20 patients requiring split-thickness skin grafts. INTERVENTION: For each patient, the donor sites were randomly assigned to be treated with meshed LSE, meshed autograft, or a polyurethane film (PUF) occlusive dressing. Blood and biopsy samples were taken for immunologic and histological studies. MAIN OUTCOME MEASURES: Toxic effects or clinically apparent rejection, humoral and cellular immune responses, clinical take, healing time, pain, and 1-month histological appearance. RESULTS: There was no toxic effect or clinically apparent rejection of LSE. Results of humoral and cellular studies were unchanged from baseline. The average time to healing for LSE with clinical take was 7.3 days (SD, +/- 0.8 days); for autograft, 7.6 days (SD, +/- 1.1 days); and for PUF, 9.5 days (SD, +/- 1.8 days). The difference between LSE or autograft and PUF was statistically significant at the .001 level. Pain was experienced by 1 patient, no patients, and 10 patients at the LSE, autograft, and PUF sites, respectively. Histologically, LSE had the thickest epidermis (P = .02), PUF had the greatest degree of fibrosis (P = .02), and autograft had the least degree of increased inflammation (P = .004) and vascularity (P = .01). CONCLUSIONS: In acute donor site wounds, LSE appeared to clinically take and to be a safe and usable form of tissue therapy. PMID- 10456340 TI - Risk factors associated with the failure of a venous leg ulcer to heal. AB - BACKGROUND: Venous leg ulcers afflict a significant portion of the population. The most popular form of therapy for venous leg ulcers is a compression bandage (eg, Unna boot), a therapy that is frequently unsuccessful. OBJECTIVE: To describe risk factors associated with the failure of a wound to heal when treated with a limb-compression bandage for 24 weeks. DESIGN: A retrospective cohort study. SETTING: Single-center outpatient specialty clinic at an academic medical center. PARTICIPANTS: Two hundred sixty consecutive patients with chronic venous leg ulcers. MAIN OUTCOME MEASURE: The magnitude of the effect of a given risk factor on the probability that a wound will heal within 24 weeks of care. RESULTS: Based on an assessment of leg wounds during initial office visits, we observed that the failure of a wound to heal within 24 weeks was significantly associated with larger wound area, measured in square centimeters (odds ratio [OR], 1.19; 95% confidence interval [CI], 1.11-1.27), duration of the wound in months (OR, 1.09; 95% CI, 1.04-1.16), history of venous ligation or venous stripping (OR, 4.58; 95% CI, 1.84-11.36), history of hip or knee replacement surgery (OR, 3.52; 95% CI, 1.12-11.08), ankle brachial index of less than 0.80 (OR, 3.52; 95% CI, 1.12-11.08), and the presence of fibrin on more than 50% of the wound surface (OR, 3.42; 95% CI, 1.38-8.45). CONCLUSIONS: Several risk factors are associated with the failure of a patient's venous leg ulcer to heal while using limb-compression therapy. It is prudent to consider these factors when referring a patient to a wound care subspecialists or for alternative therapies. PMID- 10456341 TI - Oral and genital ulceration: a unique presentation of immunodeficiency in Athabascan-speaking American Indian children with severe combined immunodeficiency. AB - BACKGROUND: Oral and genital ulcerations have been previously reported in 3 Navajo children diagnosed with severe combined immunodeficiency disease with T- and B-cell lymphopenia (T-B(-)-SCID). OBJECTIVE: To evaluate the occurrence of oral and genital ulcerations in 12 Athabascan-speaking American Indians with a diagnosis of T-B(-)-SCID (SCIDA group) and to compare their occurrence in non Athabascan-speaking children with SCID (control group). We also observed the course of these ulcerations in response to bone marrow transplantation (BMT). DESIGN: Retrospective survey of the medical records of patients with SCID admitted from December 1, 1986, through July 31, 1995. SETTING: Pediatric Bone Marrow Transplantation Division at a university hospital. PATIENTS: Twelve children in the SCIDA group and 21 in the control group. All patients had virtual absence of T- and B-cell numbers and function at time of diagnosis. RESULTS: Oral and/or genital ulcers developed as a presenting feature of the SCIDA group. These ulcerations were not observed in the 21 controls. All patients underwent BMT. Of the 10 patients with oral and/or genital ulcerations, 3 had poor T-cell reconstitution after BMT, with recurrences of ulcers requiring additional BMTs. CONCLUSIONS: Oral and/or genital ulcerations are common in Athabascan-speaking American Indian children with T-B(-)-SCID but are not seen in non-Athabascan speaking children with SCID. Thus, oral and/or genital ulceration appears to be an important, distinctive finding, and often a presenting feature of immunodeficiency in Athabascan-speaking American Indian children with SCID. Bone marrow transplantation with successful T-cell engraftment appears to be curative in the resolution of the ulcers, with recurrences only in patients who had poor T cell reconstitution. PMID- 10456342 TI - Melanoma prevention: evaluation of a health education campaign for primary schools. AB - OBJECTIVE: To evaluate the effectiveness of an educational campaign aimed toward limiting children's sun exposure. DESIGN: Prospective, multicenter trial with before-after comparison. A school-based campaign was presented as a game during a 4-week period (May 25 to June 1992) with primary school teachers as game hosts. Children were interviewed with a standardized questionnaire in September before (1991) and after (1992) the campaign. Comparisons between the children's answers before and after the campaign were made using paired chi2 tests and analysis of variance. SETTING: Five French primary schools. SUBJECTS: All children in their fourth year of primary school (228 children aged 9 years). MAIN OUTCOME MEASURES: Changes after the campaign in children's answers concerning their knowledge, attitude, and behavior toward the sun during summer holidays. RESULTS: Compared with the precampaign answers, more children after the campaign claimed to protect themselves from the sun with a hat (33.7% vs 23.8%; P = .01) or sunscreen (34.8% vs 25.4%; P = .03), avoided going outside during the sunniest hours (76.8% vs 66.0%; P = .02), reapplied sunscreens (22.1% vs 10.6%; P<.001), considered that a T-shirt and shade provided better protection than sunscreen (82.7% vs 74.8%; P = .05), considered sunlight as a risk factor for skin cancer (74.9% vs 50.7%; P<.001), and spent significantly less time in the sun with their arms (P = .005), trunk, legs, and head uncovered (P<.001). Children with a fair complexion, who were the target of this campaign, showed the best improvement in their responses. CONCLUSION: Health education campaigns can be effective in terms of improving the knowledge, attitude, and behavior of young children. PMID- 10456343 TI - Desmoglein 1 and desmoglein 3 are the target autoantigens in herpetiform pemphigus. AB - OBJECTIVE: To determine the cell surface autoimmune target of herpetiform pemphigus (HP). DESIGN: Serum samples of HP were examined by immunoblot studies with human epidermal extracts, enzyme-linked immunosorbent assay with baculovirus expressed recombinant desmoglein (rDsg) 1 and rDsg3, and immunoadsorption assay with rDsg. PATIENTS: Twenty serum samples were obtained from patients with HP who have typical clinical and histological features. All serum samples showed positive staining against keratinocyte cell surfaces by indirect immunofluorescence studies with healthy human skin. RESULTS: Immunoblot results showed that of 17 HP serum samples, only 5 reacted with a 160-kd band and 1 reacted with a 130-kd band. Results of enzyme-linked immunosorbent assays with rDsg1 and rDsg3 demonstrated that of 20 HP serum samples, 16 were positive against Dsg1 and 4 were positive against Dsg3. No serum samples reacted with both. Furthermore, in 19 of 20 HP serum samples, immunoreactivity against keratinocyte cell surfaces was completely removed by preincubation with rDsg1 and rDsg3 as shown by indirect immunofluorescence, excluding a possibility that these HP sera contain autoantibodies against other cell surface molecules. CONCLUSIONS: Dsg1 and Dsg3 are the major cell surface target molecules of HP, suggesting that most cases of HP are clinical variants of pemphigus foliaceus and that the rest might be variants of pemphigus vulgaris. PMID- 10456344 TI - Classic and near-classic articles in the dermatologic literature. AB - BACKGROUND: Only a small fraction of articles published in the dermatologic literature are cited frequently. Articles cited at least 100 times are known as "citation classics" and have been previously discussed. The nature of dermatologic articles cited fewer than 100 times but still much more frequently cited than average has not been clearly elucidated. OBJECTIVE: To identify the source, authorship, and citation history of original articles published in 24 dermatologic journals that were cited more than 25 times. DESIGN: We analyzed a citation database provided by the Institute for Scientific Information. We identified the journal of publication, authorship, and country of authorship for all original articles cited at least 25 times and published from 1982 to 1996 in 24 dermatologic journals. MAIN OUTCOME MEASURE: Source of most frequently cited dermatologic articles. RESULTS: Only 2139 (6.45%) of all original articles were cited more than 25 times. Articles published in 4 of 24 journals accounted for the most frequently cited articles. Only 18 (0.13%) of more than 13,500 first authors published at least 5 articles that were cited more than 25 times. CONCLUSIONS: Few articles are cited at least 25 times, and few persons are first authors of multiple articles in the dermatologic literature cited at least that frequently. PMID- 10456345 TI - Mucosal morbidity in patients with epidermolysis bullosa acquisita. AB - BACKGROUND: Epidermolysis bullosa acquisita is an acquired inflammatory and/or dermolytic subepidermal blistering disease characterized by IgG autoantibodies to type VII collagen. Four patients with documented epidermolysis bullosa acquisita were evaluated by a multidisciplinary team of care providers (4 dermatologists, an ophthalmologist, a radiologist, a voice and speech specialist, and an otolaryngologist) for 1 to 5 years to characterize mucosal involvement and its complications and response to treatment. Patients were evaluated clinically and by slitlamp examinations, endoscopies, computed tomographic scans, and videofluorographic swallowing studies. Spiral computed tomographic scans for virtual endoscopy were used for the nontraumatic evaluation of airways in 2 patients with respiratory tract compromise. OBSERVATIONS: Involvement of 5 or more mucosal sites--mouth, nose, conjunctiva, pharynx, and larynx--was documented in all patients. Complications included ankyloglossia, periodontal disease, scarring and crusting of nasal mucosa, symblepharon formation, obstruction of nasolacrimal ducts, deformation of the epiglottis, impaired phonation, dysphagia, esophageal strictures, and supraglottic stenosis requiring emergency tracheostomy. CONCLUSIONS: Epidermolysis bullosa acquisita may extensively (or predominantly) affect mucosal epithelia in a manner resembling cicatricial pemphigoid. Mucosal disease in these patients is often subclinical, can lead to serious complications, and is best managed using a multidisciplinary approach. PMID- 10456346 TI - Bone densities in patients receiving isotretinoin for cystic acne. AB - BACKGROUND: Few studies have been done of bone densities in humans receiving retinoids, despite a substantial amount of literature concerning retinoid-induced osteoporosis in animals. We prospectively measured bone density and calcium metabolism in young men (aged 17-25 years) receiving oral isotretinoin for cystic acne and in a group of healthy volunteers (aged 19-26 years). OBSERVATIONS: Compared with that in healthy control subjects, mean bone density was lower at all sites (spine, femoral neck, and Ward triangle) and was considerably more variable at the spine in young men with cystic acne even before treatment. Bone density at the Ward triangle decreased a mean of 4.4% (P = .03) after 6 months of isotretinoin use (1 mg/kg of body weight). Four patients showed decreased density of more than 9% at the Ward triangle. The difference between the mean change in bone density in the patient group and in the control group was significant at the Ward triangle (P = .04) but not at the other sites. Measurements of calcium metabolism did not change over time in either group. CONCLUSIONS: A loss of bone density occurring in the absence of measurable alterations of calcium metabolism is likely to be a direct effect of retinoids on bone. Further study of retinoid induced osteoporosis in humans and of bone density in patients with cystic acne is needed. PMID- 10456348 TI - Tissue-engineered skin: an alternative to split-thickness skin grafts? PMID- 10456347 TI - Hair follicle biology, the sebaceous gland, and scarring alopecias. PMID- 10456349 TI - Tetracycline and epidermolysis bullosa simplex: a new indication for one of the oldest and most widely used drugs in dermatology? PMID- 10456350 TI - A draining tumor in the popliteal fossa. PMID- 10456351 TI - Symmetrical hemorrhagic bullae in an immunocompromised host. PMID- 10456352 TI - Diffuse painless ulcerations. PMID- 10456353 TI - An ulcerated nodule associated with lymphadenopathy. PMID- 10456354 TI - Finasteride, 1 mg (Propecia), is the optimal dose for the treatment of men with male pattern hair loss. PMID- 10456355 TI - Finasteride in a 1-mg dose is safe and effective. PMID- 10456356 TI - 7-5 by '05. PMID- 10456357 TI - Physicians' responsibility for medical and surgical dermatology. PMID- 10456358 TI - Wound measurement made truly simple by point counting. PMID- 10456359 TI - Computers or simple wound measurements: when Greek meets Greek, then comes the tug-of-war! PMID- 10456360 TI - Congenital cutaneous defects in co-twins. PMID- 10456361 TI - The American Academy of Dermatology should not endorse product sales from physicians' offices. PMID- 10456362 TI - Effectiveness of photopheresis in Sezary syndrome. PMID- 10456363 TI - Two familial cases of epidermolysis bullosa simplex successfully treated with tetracycline. PMID- 10456364 TI - Multivariate analysis in alopecia areata: risk factors and validity of clinical forms. PMID- 10456365 TI - High prevalence of cutaneous reactions to interferon alfa plus ribavirin combination therapy in patients with chronic hepatitis C virus. PMID- 10456366 TI - Hepatitis C virus infection in cutaneous polyarteritis nodosa: a retrospective study of 16 cases. PMID- 10456367 TI - Individual variations in allergic contact dermatitis from urushiol. PMID- 10456368 TI - Metabolic and functional effects of low-potassium cardioplegic solutions for long term heart preservation. AB - Cardioplegic solutions used to arrest the heart during open heart surgery and cardiac transplantation are based on potassium as a cardioplegic agent in a concentration range of 15-35 mM. However, high to moderate K+ concentrations increase Ca2+ influx and impair endothelial function. We have therefore evaluated the possible advantage of a lower potassium concentration in a new cardioplegic solution (named CRMBM solution) designed for long-term heart preservation. Nine isolated perfused rat hearts were submitted to 8 h of hypothermic ischemia after cardioplegic arrest, followed by 60 min of reflow at 37 degrees C. Two cardioplegic solutions were compared: (1) the CRMBM solution with 10 mM potassium (K-10 group), and (2) the CRMBM solution with 4 mM potassium (K-4 group). The quality of heart preservation was assessed by a metabolic study using P-31 magnetic resonance spectroscopy (energy metabolism and intracellular pH) combined to a functional evaluation and a measure of cellular integrity (biochemical assays in effluents and tissues). Decreasing the potassium concentration to 4 mM improved heart preservation, as shown by a higher functional post-ischemic recovery represented by the rate pressure product and a better preservation of cellular integrity. The evolutions of intracellular pH and high energy phosphate levels during ischemia and reflow were similar in both groups. PMID- 10456369 TI - Magnetic resonance imaging of rodent teeth. AB - In this paper, the anatomical structures of rat jaws are studied using magnetic resonance imaging (MRI) with high spatial resolution. In vivo microimages of teeth from 3- and 12-week-old (young and adult) rats have been performed. A 2 T nuclear magnetic resonance microscope was used to collect, with multiple orientations, non-invasive 2D data images using the spin-echo technique. MRI appears well suited to give images of the oral area and may represent a useful tool for diagnosis of dental diseases and more particularly of pulp pathologies. PMID- 10456370 TI - Magnetic resonance imaging of the pituitary gland in patients with secondary hypogonadism due to transfusional hemochromatosis. AB - To identify pituitary iron overload in patients with transfusional hemochromatosis causing secondary hypogonadism, we prospectively evaluated signal intensity abnormalities of the anterior lobe of the pituitary gland of 18 patients affected by transfusion-dependent thalassemia major and secondary hypogonadism. Magnetic resonance (MR) imaging is useful to assess pituitary iron overload in patients with transfusional hemochromatosis and secondary hypogonadism by detection of a significant decreased signal intensity of the anterior lobe of the pituitary gland on GRE T2*-weighted images. The decreased signal intensity of the anterior lobe of the pituitary gland on GRE T2*-weighted images was correlated to increasing serum ferritin level (r = -0.84, r2 = -0.70, P < 0.001). Indeed, the lower the signal intensity of the pituitary gland, the greater the serum ferritin level. However an exact quantification of pituitary iron overload by correlation with serum ferritin level is not allowed. No correlation was found between MR imaging results and hormonal status; however, the detection of pituitary iron overload on GRE T2*-weighted images is consistent with the hypothesis of hypogonadotrophic pituitary insufficiency due to iron induced cellular damage. PMID- 10456371 TI - Endorectal coil MR imaging findings in hemospermia. AB - The purpose of this study was to illustrate the spectrum of abnormalities found at endorectal coil magnetic resonance (MR) imaging in patients with hemospermia. Ninety patients aged between 23 and 71 years (mean age 41 years) presenting with hemospermia underwent evaluation with endorectal coil MR imaging at 1.5 T. Duration of hemospermia ranged between 5 days and 4 years (mean 15 months). MR examination protocol included T1-weighted spin-echo (SE) and T2-weighted fast SE MR images obtained in the sagittal, coronal, and axial planes. Abnormalities were observed on endorectal-coil MR images in 49 of 90 patients (54%). Blood within the seminal vesicle or the ejaculatory duct was recognized in 23 of 90 patients (25%). Dilatation of the seminal vesicles or the ejaculatory duct was observed in 31 of 90 patients (34%). Cystic lesions were identified in 14 cases, eight of which involved the utriculus and six the ejaculatory duct. Calculi within the seminal vesicles were depicted in seven patients. No malignant disease was demonstrated. In conclusion, despite the evidence that hemospermia is most often a benign and self-limiting condition, we found that endorectal coil MR imaging can detect abnormal findings in more than half of the patients, and may be helpful in assessing the level at which hemorrhage occurred and in defining the cause of the disease. PMID- 10456372 TI - Water diffusion in rat brain in vivo as detected at very large b values is multicompartmental. AB - The diffusion-weighted signal attenuation of water in rat brain was measured with pulsed-field gradient nuclear magnetic resonance methods in a single voxel under in vivo and global ischemic conditions. The diffusion-attenuated water signal was observed in vivo at b values of 300 ms/microm2 (strength of diffusion weighting) and diffusion times up to 400 ms. A series of constant diffusion time (CT) experiments with varied gradient directions and diffusion times revealed a multiexponential decay with apparent diffusion coefficients (ADC) covering two orders of magnitude from 1 to 0.01 microm2/ms. In a four-exponential fit, the observed changes during global ischemia could be fully explained by changes in the relative volume fractions only with unchanged ADCs. An anisotropy of the ADC, detected at small b values, was not observed for the ADC at large b values, but for the concomitant volume fractions. An inverse Laplace Transform of the CT curves, performed with CONTIN, resulted in continuously distributed diffusion coefficients, for which the term 'diffusogram' is proposed. This approach was more appropriate than a discrete exponential model with four to six components, being related to the morphology of brain tissue and its cell size distribution. On the basis of an analytical, quantitative model, it is suggested that the measured ADC at small b values reflects mainly properties of the restricting boundaries, i.e. the relative volume fractions and the extracellular tortuosity, while the intrinsic intracellular diffusion constant and the exchange time are predicted to have minor influence. PMID- 10456373 TI - 1H spectroscopic imaging of acute head injury--evidence of diffuse axonal injury. AB - Using single slice two-dimensional spectroscopic imaging (SI), nine acute head injury patients and six controls have been successfully scanned. The problems presented by the need for ITU monitoring of these patients during MR scanning was overcome using MR compatible monitoring equipment. In previous studies of head injury which used proton spectroscopy, single voxel localisation procedures have meant that the spatial extent of the spectral data has been limited. With spectral data from a whole axial slice, we have been able to identify NAA abnormalities in regions remote to any T2 visible lesions. This suggests that SI (of NAA in particular) will be useful for the diagnosis of diffuse axonal injury. PMID- 10456374 TI - Gradiometer pick-up coil design for a low field SQUID-MRI system. AB - We describe the use of liquid helium-cooled (4.2 K) gradiometer coils and a DC superconducting quantum interference device (SQUID) preamplifier to improve the SNR of magnetic resonance imaging (MRI) at 0.01 T. Gradiometer windings are used both to reduce lossy interactions with the MRI system's room temperature magnet and gradient coils and also to reject interference from more distant sources, which reduces the need for RF shielding. We have tested both axial and planar (figure-of-eight) gradiometer configurations. The figure-of-eight gradiometer has a more rapid fall-off in sensitivity with increasing distance from its windings than the axial gradiometer, but this is compensated for by reduced lossy interactions and improved interference rejection. We have used the system to image the human arm. PMID- 10456375 TI - Comparison of in vivo 1H MRS of human brain tumours with 1H HR-MAS spectroscopy of intact biopsy samples in vitro. AB - High resolution magic angle spinning (MAS) 1H nuclear magnetic resonance (NMR) spectroscopy has been employed to study intact human brain tumour tissue and comparison with the corresponding in vivo spectrum has been made. Two dimensional 1H MAS-NMR measurements, including J-resolved and homonuclear shift correlation spectra, were obtained to aid metabolite signal assignment. MAS gave greatly improved line-shape and reduced line-width in comparison to conventional high resolution in vivo 1H MRS of intact tissue, permitting the simultaneous detection of cellular lipids and metabolites. The technique provides the most direct method for comparison of in vivo spectra with high resolution spectra in vitro and hence allows more reliable peak assignment of in vivo 1H MRS spectra. PMID- 10456376 TI - Biological interpretation of relative risk. AB - There is widespread interest in assessing the clinical importance of a study result. This goal is impeded, however, by a lack of clarity about the biological interpretability of epidemiological effect measures, such as the relative risk. A relative risk is often interpreted merely as a measure of some vague statistical association, without a view toward a biological effect as an object of measurement. Not infrequently, if it is not statistically significant, the relative risk estimate is ignored completely. A key to biological interpretation is appreciating the theoretical framework stipulating that outcome rates derived from 2 comparison groups actually represent measures of different effects in the same population. For instance, by using a placebo group to estimate the number of background cases that occurred in the treatment group, an estimate of the number of excess cases that occurred as a result of treatment can be made. This kind of biological entity can be derived from a relative risk, and can be more easily evaluated as to its clinical importance than a statistical association or a statement about statistical significance. Interpretation then becomes a more directed task, with a focus on the validity of certain ancillary hypotheses upon which biological interpretability rests. PMID- 10456377 TI - Use of phenothiazines as sedatives in children: what are the risks? AB - Phenothiazines have been widely used for their antiemetic, antipsychotic and sedative properties for many years. The introduction of alternative agents for paediatric sedation has led to the re-evaluation of phenothiazines as paediatric sedatives. Newer agents, such as fentanyl and midazolam, have short half-lives and reversal agents are available. Therefore these agents may offer comparable therapeutic efficacy with a better safety profile in young children. Reports of sudden infant death syndrome in children receiving a phenothiazine-containing syrup for symptoms of upper respiratory infection means that the outpatient use of these compounds in very young infants is not recommended. PMID- 10456378 TI - Drug-induced lymphopenia: focus on CD4+ and CD8+ cells. AB - Drug-induced lymphopenia is a common adverse event. Some drugs, in particular those used in the treatment of malignancies and autoimmune diseases, inevitably affect the percentages and proportions of lymphocytes in the peripheral blood. Some other drugs exert only minor effects and their clinical relevance cannot be established with certainty. Most cytotoxic and immunosuppressive drugs affect CD4+ T cells more profoundly. Since their regeneration seems to be slower than that of CD8+ T cells, the frequent occurrence of CD4+ lymphopenia may merely reflect this phenomenon. As in HIV infection, critically low numbers of CD4+ cells, irrespective of the cause, predisposes to opportunistic infections. There is no such critically low value for CD8+ cells, and their essential role in various pathological conditions should also be established. PMID- 10456380 TI - Potential interactions of the extended-spectrum fluoroquinolones with the CNS. AB - The new generation fluoroquinolones -- sparfloxacin, levofloxacin, grepafloxacin and trovafloxacin -- have been designed to respond to the clinical need for extended antimicrobial cover in the face of increasing global microbial resistance. Their main focus is in the treatment of respiratory infections, particularly those acquired in the community. CNS adverse effects, such as dizziness and headache, are known to occur relatively commonly with some fluoroquinolones and are not, in general, well tolerated by patients. The structural component of the fluoroquinolone molecule believed to be responsible for improved gram-positive activity is also believed to be implicated in the production of CNS adverse effects, including those arising from drug interactions with theophylline and NSAIDs. Inhibition of brain gamma-aminobutyric acid (GABA) receptor binding appears to be a strong indicator of CNS activity, though N methyl-D-aspartate receptor binding has also been implicated. In accordance with the results of these predictive studies, clinical trials have found sparfloxacin, levofloxacin and grepafloxacin to be associated with a low incidence of CNS events. Trovafloxacin has been found to be associated with a higher incidence of CNS events (particularly lightheadedness and dizziness) than the other 3 agents. Ongoing and future clinical studies will help to define the usefulness of the predictive models, as well as reveal the full CNS adverse event profile of these and other investigational fluoroquinolones. PMID- 10456379 TI - Drug-induced cognition disorders in the elderly: incidence, prevention and management. AB - The aetiology of cognitive impairment is multifactorial; however, drugs are an important cause of delirium and dementia. Several factors may increase the risk of drug-induced cognition disorders in the elderly including imbalances in neurotransmitters (e.g. acetylcholine), age-related alterations in pharmacokinetics and pharmacodynamics, and high levels of medication use. Nearly any drug can cause cognitive impairment in susceptible individuals; however, certain classes are more commonly implicated. Benzodiazepines, opioids, anticholinergics, and tricyclic antidepressants are probably the worst offenders. Older antihypertensive agents (reserpine, clonidine) have negative effects on cognition; however, large clinical trials in the elderly indicate that commonly used agents [e.g. thiazide diuretics, calcium antagonists (amiodipine, diltiazem), ACE inhibitors (captopril, enalapril) and beta-blockers (atenolol)] have minimal effects on cognition. Newer antidepressants such as selective serotonin (5-hydroxytryptamine; 5-HT) reuptake inhibitors (SSRIs) and reversible inhibitors of monoamine oxidase A have not been shown to have negative effects on cognition. Although some drugs have shown low risk for causing cognition disorders in research studies, risk may be increased in frail older adults taking several medications and each case should be reviewed carefully. Identification of drug-induced cognitive impairment is crucial to early detection and resolution of symptoms. Preventive strategies directed at avoiding high risk medications when possible, appropriately adjusting doses based on age-related changes and close follow-up may prevent these conditions. PMID- 10456381 TI - Risk-benefit assessment of therapies for Mycobacterium avium complex infections. AB - Mycobacterium avium complex (MAC) is an important pathogen that can cause chronic lung disease in immunocompetent patients and disseminated disease in patients with the acquired immunodeficiency syndrome (AIDS). Treatment of MAC with antituberculosis drugs was unsatisfactory, but the introduction of the newer macrolides, clarithromycin and azithromycin, and of rifabutin has greatly improved the outcome of treatment regimens for MAC. However, these agents are also associated with many new treatment-related adverse effects and potential drug-drug interactions. Rifamycins [rifampicin (rifampin) more than rifabutin] induce cytochrome P450 enzymes and accelerate the metabolism of clarithromycin and HIV protease inhibitors. Conversely, clarithromycin inhibits these enzymes, resulting in increased rifabutin toxicity. The net results are treatment regimens that may be extremely difficult to tolerate, especially for elderly or debilitated patients. Clarithromycin and azithromycin must be administered in combination with other agents such as ethambutol to prevent the emergence of macrolide resistance. Unfortunately, not all patients respond to the combination of a macrolide, rifabutin and ethambutol, and many have significant adverse effects (mostly gastrointestinal) with this regimen. For some patients the treatment is worse than the disease. The same 3-drug regimen is also effective therapy for disseminated MAC in AIDS patients, in whom the additional problem of a rifamycin/protease inhibitor interaction may be present. Fortunately, as opposed to pulmonary MAC disease in immunocompetent patients, disseminated MAC disease is a diminishing problem because of effective prophylactic regimens for MAC and improved antiretroviral therapy for HIV. Significant progress has been made in the treatment of MAC disease with the introduction of the newer macrolides. It is to be hoped that even better drugs that are more active against MAC and are associated with less toxicity and drug-drug interactions will be introduced in the future. PMID- 10456382 TI - The toothless osteopetrotic rat has a normal vitamin D-binding protein-macrophage activating factor (DBP-MAF) cascade and chondrodysplasia resistant to treatments with colony stimulating factor-1 (CSF-1) and/or DBP-MAF. AB - The osteopetrotic rat mutation toothless (tl) is characterized by little or no bone resorption, few osteoclasts and macrophages, and chondrodysplasia at the growth plates. Short-term treatment of tl rats with colony-stimulating factor-1 (CSF-1) has been shown to increase the number of osteoclasts and macrophages, producing dramatic resolution of skeletal sclerosis at some, but not all, sites. Defects in production of vitamin D-binding protein-macrophage activating factor (DBP-MAF) have been identified in two other independent osteopetrotic mutations of the rat (op and ia), and two in the mouse (op and mi), in which macrophages and osteoclasts can be activated by the administration of exogenous DBP-MAF. The present studies were undertaken to examine the histology and residual growth defects in tl rats following longer CSF-1 treatments, to investigate the possibility that exogenous DBP-MAF might act synergistically with CSF-1 to improve the tl phenotype, and to assess the integrity of the endogenous DBP-MAF pathway in this mutation. CSF-1 treatment-with or without DBP-MAF-induced resorption of metaphyseal bone to the growth plate on the marrow side, improved slightly but did not normalize long bone growth, and caused no improvement in the abnormal histology of the growth plate. Injections of lysophosphatidylcholine (lyso-Pc) to prime macrophage activation via the DBP-MAF pathway raised superoxide production to similar levels in peritoneal macrophages from both normal and mutant animals, indicating no defect in the DBP-MAF pathway in tl rats. Interestingly, pretreatments with CSF-1 alone also increased superoxide production, although the mechanism for this remains unknown. In summary, we find that, unlike other osteopetrotic mutations investigated to date, the DBP-MAF pathway does not appear to be defective in the tl rat; that additional DBP-MAF does not augment the beneficial skeletal effects seen with CSF-1 alone; and that the growth plate chondrodystrophy seen in this mutation is unaffected by either molecule. Thus, the tl mutation intercepts the function of a gene required for both normal endochondral ossification and bone resorption, thereby uncoupling the coordination of skeletal metabolism required for normal long bone growth. PMID- 10456383 TI - Cortical tibial bone volume in two strains of mice: effects of sciatic neurectomy and genetic regulation of bone response to mechanical loading. AB - Although C3H/HeJ (C3H) and C57BL/6J (B6) mice are similar in body size (and adult weight), and have bones of similar external size, C3H mice have higher peak bone densities than B6 mice (e.g., 53% higher peak bone density in the femora). The current studies were intended to assess the role of mechanical loading/unloading as a possible determinant of the bone density difference between these inbred strains of mice and, specifically, to assess the effect of sciatic neurectomy on histomorphometric indices of bone formation and resorption in the tibiae of female C3H and B6 mice. Groups of 10 mice of each strain were subjected to left side sciatic neurectomy (left hindlimb immobilization) or a sham procedure. The contralateral (right) legs of each mouse were used as controls. Four weeks of immobilization produced no systemic changes in bone formation indices in either strain of mice (i.e., no change in serum alkaline phosphatase or serum osteocalcin). However, histomorphometric assessments at the tibiofibular junction showed that 4 weeks of immobilization caused a time-dependent decrease in the length of the endosteal bone forming perimeter (e.g., 14% of control single labeled, noneroded surface at 4 weeks, p < 0.005) with a concomitant increase in the length of the endosteal bone resorbing perimeter (i.e., 424% of control eroded surface at 4 weeks, p < 0.005), in the B6 mice. These effects were associated with an increase in medullary area (132% of control, p < 0.05) at this site, in the B6 mice. The pattern of response was different in the tibiae of the C3 mice-a much smaller decrease in bone forming perimeter (88% of control at 4 weeks, p < 0.05), with no associated increase in bone resorbing perimeter, and no change in medullary area. Similar effects were seen at a second cross-sectional sampling site, in the proximal tibia. Together, these findings indicate that B6 mice are more sensitive to endosteal bone loss from hindlimb immobilization than C3H mice. PMID- 10456384 TI - Local administration of activin promotes fracture healing in the rat fibula fracture model. AB - Osteogenic activities of activin, a member of the transforming growth factor (TGF)-beta superfamily, have been demonstrated in both in vitro and in vivo studies. The present study investigates the effects of topical application of activin on fracture healing using rat fibula fracture models. Activin (0.4-10 microg/day) was injected locally to the fracture once a day for 2 weeks. Activin promoted callus formation in a dose-dependent manner and both callus volume and callus weight were significantly increased with doses of 2-10 microg/day activin. Also, 3 weeks of activin treatment increased the mechanical strength of the healing bone in addition to the callus mass. Histological study 2 weeks after the fracture revealed that activin promoted endochondral bone formation. Immunohistochemical examination of the fractured tibia revealed that activin was localized to osteoblasts and chondrocytes in the region ossified both endochondrally and intramembranously. These findings suggest that activin expressed during fracture healing promotes the healing process through an autocrine/paracrine mode of action. PMID- 10456385 TI - Expression of metalloproteinase-13 (Collagenase-3) is induced during fracture healing in mice. AB - In fracture healing, a large amount of cartilage is formed, then rapidly replaced by osseous tissue. This process requires the transition of extracellular matrix component from type II to type I collagen. We investigated the expression of matrix metalloproteinase-13 (MMP-13), which has a high potential to cleave type II as well as type I collagen, during fracture repair in mouse ribs. In situ hybridization demonstrated that MMP-13 mRNA was present throughout the healing process. It was detected in the cells of the periosteum at day 1. As fracture callus grew, strong MMP-13 mRNA signals were detected in cells of the cartilaginous callus. In the reparative and remodeling phases, both hypertrophic chondrocytes and immature osteoblastic cells in the fracture callus expressed MMP 13 mRNA strongly. These cells were located adjacent to tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts at the sites of cartilage/bone transition. In osteoclasts, MMP-13 expression was not detected. The level of MMP 13 mRNA peaked at day 14 postfracture by northern blotting. Immunohistochemical staining showed that MMP-13 was detected primarily in hypertrophic chondrocytes. These results indicate that MMP-13 is induced during fracture healing. The site- and cell-specific expression of MMP-13 and its enzymatic property suggest that MMP-13 initiates the degradation of cartilage matrix, resulting in resorption and remodeling of the callus. In conclusion, MMP-13 plays an important role in the healing process of fractured bone in mice. PMID- 10456386 TI - Cloning and expression of rhesus monkey cathepsin K. AB - Cathepsin K is a cysteine protease involved in degradation of human type I collagen and plays a primary role in bone resorption. We have cloned rhesus monkey cathepsin K by reverse transcriptase-polymerase chain reaction (RT-PCR) from rhesus ovary poly A+ RNA. The sequence for the rhesus enzyme is 98% identical to that of the human with 100% identity within the mature active form of cathepsin K. Rhesus monkey cathepsin K was transiently expressed in Chinese hamster ovary (CHO) cells and found to be secreted as the proenzyme in the culture media and 50% activated to the mature form intracellularly. The substrate specificity preference of aminomethylcoumarin and rhodamine peptide substrates was Leu > Phe > Pro in the P2 position when tested with constant arginine at P1. The enzyme activity expressed in CHO cell extracts was sensitive to inhibition by E-64 and cystatin with IC50s of 3.5 nmol/L and 13 ng/mL, respectively. The apparent second order rate constants of inactivation by E-64 were 66,000 M(-1) s( 1) and 130,000 M(-1) s(-1) for the recombinantly expressed rhesus monkey and human cathepsin K, respectively. The high similarity between the sequences and the kinetic properties of rhesus monkey and human cathepsin K establishes this monkey species as a suitable animal model for development of novel cathepsin K inhibitors as antiresorptive agents. PMID- 10456387 TI - Conditioned medium of estrogen-treated osteoblasts inhibits osteoclast maturation and function in vitro. AB - The increase of bone resorption and reduction of bone mass in postmenopausal women can be prevented by treatment with estrogen. Although it is well established that estrogen treatment normalizes the increased bone turnover, the mechanism by which estrogen exerts its protective influence at the cellular and molecular level in bone remains elusive. It has been shown that osteoblasts are involved in osteoclast development and osteoclastic bone resorption. In this work we examine the effect of estrogen (E2) on osteoclast-mediated bone resorption via the medium conditioned by osteoblast cultures. The conditioned medium collected from osteoblast cultures without (CM) or with 0.1 nmol/L 17beta-estradiol (E-CM) was mixed in a 1:1 ratio with fresh osteoclast culture medium. Osteoclasts were isolated from the bone marrow of 3-day-old NMRI mice and cultured on bovine bone slices. The total number of multinucleated tartrate-resistant alkaline phosphatase (TRAP)-positive cells in cultures with CM and E-CM was similar to that of cells incubated in control medium. However, the number of osteoclasts containing more than three nuclei was significantly smaller in the cultures containing E-CM. The total area of resorption was only slightly decreased in cultures containing CM, but was markedly inhibited in cultures with E-CM. In osteoblast cultures, the production of interleukin (IL)-1 and IL-6, but not of TNF-alpha, was reduced by 0.1 nmol/L E2. Our data suggest that E2 treatment of osteoblasts decreases the production of factor(s) that induces osteoclast differentiation to multinucleated cells with a higher capacity for bone resorption. PMID- 10456388 TI - Microdamage adjacent to endosseous implants. AB - Intense remodeling occurs in lamellar bone adjacent to osseointegrated endosseous implants. The purpose of this study was to compare microdamage accumulation subsequent to ex vivo fatigue loading of bone that surrounds an endosseous implant, (a) immediately after placement (nonadapted bone) and (b) following a 12 week healing period after placement (adapted bone). We hypothesize that there is less microdamage in the more compliant adapted bone than in the older nonadapted bone. Nonthreaded titanium plasma sprayed (TPS)-coated endosseous implants were placed into dog mid-femoral diaphyses and allowed to heal for 12 weeks. Block sections of bone, each containing one implant, were cut anteroposteriorly, resulting in an implant containing lateral cortex, and a medial cortex that was used for testing the nonadapted specimens. Control specimens (n = 14 each for adapted and nonadapted) were loaded at 0 N. Experimental specimens (n = 13, adapted; n = 14, nonadapted) were loaded at 100 N in cantilever bending for 150,000 cycles at 2 Hz, at 37 degrees C on a Bionix 858 testing machine. Specimens were bulk stained with basic fuchsin and 120-140 microm sections were obtained. Crack numerical density (Cr.Dn = Cr.N/ B.Ar, #/mm2), crack surface density (Cr.S.Dn = Tt.Cr.Le/ B.Ar, mm/mm2), and percent damage area (Dm.Ar = Cr.Ar x 100/B.Ar, mm2/mm2) were measured at x 250. Statistically significant differences (p < 0.0001) were seen for Cr.Dn, Cr.S.Dn, and Dm.Ar on the compressed cortices suggesting that adapted bone near the implant accumulated significantly less microdamage than nonadapted bone. Also, the adapted nonloaded control specimens had approximately 20-fold less damage than the respective nonadapted specimens. This study suggests that the compliant adapted bone adjacent to endosseous implants is relatively resistant to fatigue loads. The high success rates of endosseous implants may be due to the presence of a rapidly remodeling region that maintains tissue compliance and limits microdamage initiation. PMID- 10456389 TI - Biomechanical effects of intraspecimen variations in trabecular architecture: a three-dimensional finite element study. AB - Trabecular architecture is considered important in osteoporosis and has been quantified by a variety of mean parameters characteristic of a whole specimen. Variations within a specimen, however, have been mostly ignored. In this study, the theoretical effects of these intraspecimen variations in architecture on predicted mechanical properties were investigated through a three-dimensional finite element parameter study that simulated variations in trabecular thickness in a controlled manner. An irregularly spaced lattice of different sized rods was used to simulate trabecular bone in three distinct volume fraction ranges, representing young, middle-aged, and elderly vertebral bone. Beta distributions (a type of non-normal distribution) of trabecular thickness with coefficients of variation of either 25%, 40%, or 55% were applied to the rods in each model, and 225 simulations of uniaxial compression tests were performed to obtain modulus values. Percent modulus reductions of 22% and 43% were predicted when the intraspecimen coefficient of variation in trabecular thickness was increased from 25% to 40% and from 25% to 55%, respectively, for models of equal volume fraction. Furthermore, this trend was predicted to be independent of volume fraction. We conclude, therefore, that consideration of the intraspecimen trabecular thickness variation in conjunction with volume fraction may improve the ability to predict trabecular modulus compared with use of volume fraction alone. Further, the model suggests that if age, disease, or drug treatments increase trabecular thickness variation, this may be detrimental to mechanical properties. PMID- 10456390 TI - Comparison of force attenuation properties of four different hip protectors under simulated falling conditions in the elderly: an in vitro biomechanical study. AB - The purpose of this in vitro biomechanical study is to determine the force attenuation capacity of four different hip protectors (KPH1, KPH2, Safehip, and Safetypants) in falling simulations in elderly subjects (falls to the side). The simulated falling conditions were created by a biomechanical testing system, which consisted of an impact pendulum, surrogate pelvis and femur, and two load cells. Three series of impact experiments were conducted in an ascending order (low, moderate-, and high-force experiments), each exceeding the literature provided average (+/- 1 SD) fracture threshold (3100 +/- 1200 N) of the proximal femur of elderly women with a mean age of 71 years. Using a low impact force of 4330 N, the trochanteric soft tissue (20-mm-thick polyethylene foam) attenuated the peak femoral impact force to 3740 N and, accordingly, the KPH1 protector to 590 N, KPH2 to 510 N, Safehip to 1080 N, and Safetypants to 790 N. Thus, in this low force experiment, all tested protectors could reduce the peak impact force entered into the proximal femur below the aforementioned average fracture threshold area (3100 +/-1200 N) of the proximal femur of elderly women. With a moderate impact force of 7230 N, the soft tissue attenuated the peak femoral impact force to 6130 N, and the protectors to 780 N, 760 N, 2240 N, and 2760 N, respectively. Thus, with this impact force, only the KPH hip protectors could reduce the impact force clearly below the fracture threshold area. In the final series of the experiment, the peak femoral impact force was set to be so high (10,840 N) that the protector, if effective, should prevent the hip fracture in almost all cases and situations. The trochanteric soft tissue attenuated this peak impact force to 9190 N, and the tested protectors to 1360 N, 1170 N, 4640 N, and 5770 N. Thus, with the KPH protectors the force received by the proximal femur remained below the average force required to fracture the proximal femur of elderly women, whereas with the two other protectors the impact force entered into the proximal femur clearly exceeded this threshold value. In conclusion, the test results showed that, of the four tested hip protectors, the anatomically designed energy-shunting and energy-absorbing KPH protectors can provide an effective impact force attenuation in a sideways-fall simulation in the elderly, whereas the force attenuation capacity of the two other protectors seems more limited. However, the true efficacy of any protector in the prevention of hip fractures can only be evaluated in randomized clinical trials. PMID- 10456391 TI - Primary hyperparathyroidism: short-term changes in bone remodeling and bone mineral density following parathyroidectomy. AB - Changes in bone remodeling and bone mineral density were observed during a period of 6 months after surgery in 24 patients with primary hyperparathyroidism (20 women and 4 men; age 54+/-12 years, range 26-69 years). All bone markers declined significantly within the 6 month follow-up period, but the time course for changes in renal N-terminal telopeptide of type 1 collagen (NTx) excretion differed from those of the other markers by a steep and significant reduction (p < 0.05) after less than 1 month. During the 6 month period, bone mineral density (BMD) increased significantly at all sites measured (p < 0.05) apart from the femoral neck and the proximal and midforearm. The greatest increase of 4.2% was observed in the trochanteric region (p < 0.001). The increase in BMD in spine, trochanteric, and intertrochanteric regions of the hip correlated inversely with baseline forearm BMD values (p < 0.05). Baseline bone markers (serum alkaline phosphatase [AP], serum bone AP, serum pyridinoline crosslinked telopeptide of type 1 collagen, urinary hydroxyproline, urinary osteocalcin), as well as baseline histomorphometric indices of bone turnover (eroded and labeled surface, bone formation rate, activation frequency, and cortical porosity) were positively correlated with changes in spinal BMD over 6 months (p < 0.05). It was concluded that, within 6 months after parathyroidectomy, patients with primary hyperparathyroidism obtain normalization of bone remodeling and a substantial increase in bone mineral density in regions rich in cancellous bone but no significant changes in regions with predominantly cortical bone. PMID- 10456392 TI - Measurement of intraspecimen variations in vertebral cancellous bone architecture. AB - A three-dimensional technique was developed for the quantification of the number and cross-sectional geometry of individual trabeculae oriented along a given direction. As an example application, the number of vertical and horizontal trabeculae and their respective cross-sectional geometry were determined for a set of six vertebral cancellous bone specimens (L3-L4 female vertebral bodies; age range 39-63 years). Three-dimensional optical images at a spatial resolution of 20 microm were obtained using an automated serial milling technique. The thickness distributions were generally right skewed. The mean true thickness for both the vertically and horizontally oriented trabeculae showed a strong relationship with volume fraction (vertical: r2 = 0.86; p < 0.05; horizontal: r2 = 0.80; p < 0.05), and mean trabecular thickness (Tb.Th.) (vertical: r2 = 0.81; p < 0.05; horizontal: r2 = 0.72; p < 0.05). The horizontal trabeculae were greater in number and were thinner than the vertical trabeculae. The coefficient of variation of the intraspecimen vertical trabecular thicknesses ranged from 25% to 42%, and showed a weak, albeit insignificant, positive correlation with volume fraction (r2 = 0.46). The findings demonstrated substantial intraspecimen variations exist in trabecular thickness that are not related to volume fraction. Further studies are recommended to determine the potential role of such intraspecimen variations in architecture on biomechanical properties. PMID- 10456393 TI - Expression of insulin receptors in osteoclast-like cells. PMID- 10456394 TI - The development of an international surgical registry: the ECSUR project. The European Cardiac Surgical Registry. AB - OBJECTIVES: Cardiac surgery is the first of any therapeutic discipline to attempt to establish a major international registry on a patient-by-patient basis. Being the first necessarily imposes a challenge since no rules or guidelines exist. Possibilities and boundaries must be identified. METHODS: The EACTS database committee established ECSUR (The European Cardiac SUrgical Registry) with the aim of collecting patient-by-patient information from centres across Europe. It was decided to collect patient data in three areas; adult cardiac surgery, paediatric cardiac surgery and thoracic surgery. Currently, these three areas are in different stages of development, with multinational data collection now having already commenced in adult cardiac surgery. RESULTS: After just over a year of data collection ECSUR has now received data from 98 centres in 30 countries, encompassing approximately 100,000 patients. Three national cardiac surgical registries currently send their data to ECSUR, and agreement has been reached with three other countries for prospective data sharing during 1999. CONCLUSION: This paper describes how ECSUR was established, the on-going status of the project, and how the project has now developed a global perspective through a joint European and US initiative to undertake international dataset unification for surgeons worldwide, by the definition of core adult cardiac, paediatric cardiac and thoracic surgical datasets. PMID- 10456395 TI - European system for cardiac operative risk evaluation (EuroSCORE). AB - OBJECTIVE: To construct a scoring system for the prediction of early mortality in cardiac surgical patients in Europe on the basis of objective risk factors. METHODS: The EuroSCORE database was divided into developmental and validation subsets. In the former, risk factors deemed to be objective, credible, obtainable and difficult to falsify were weighted on the basis of regression analysis. An additive score of predicted mortality was constructed. Its calibration and discrimination characteristics were assessed in the validation dataset. Thresholds were defined to distinguish low, moderate and high risk groups. RESULTS: The developmental dataset had 13,302 patients, calibration by Hosmer Lemeshow Chi square was (8) = 8.26 (P < 0.40) and discrimination by area under ROC curve was 0.79. The validation dataset had 1479 patients, calibration Chi square (10) = 7.5, P < 0.68 and the area under the ROC curve was 0.76. The scoring system identified three groups of risk factors with their weights (additive % predicted mortality) in brackets. Patient-related factors were age over 60 (one per 5 years or part thereof), female (1), chronic pulmonary disease (1), extracardiac arteriopathy (2), neurological dysfunction (2), previous cardiac surgery (3), serum creatinine >200 micromol/l (2), active endocarditis (3) and critical preoperative state (3). Cardiac factors were unstable angina on intravenous nitrates (2), reduced left ventricular ejection fraction (30-50%: 1, <30%: 3), recent (<90 days) myocardial infarction (2) and pulmonary systolic pressure >60 mmHg (2). Operation-related factors were emergency (2), other than isolated coronary surgery (2), thoracic aorta surgery (3) and surgery for postinfarct septal rupture (4). The scoring system was then applied to three risk groups. The low risk group (EuroSCORE 1-2) had 4529 patients with 36 deaths (0.8%), 95% confidence limits for observed mortality (0.56-1.10) and for expected mortality (1.27-1.29). The medium risk group (EuroSCORE 3-5) had 5977 patients with 182 deaths (3%), observed mortality (2.62-3.51), predicted (2.90-2.94). The high risk group (EuroSCORE 6 plus) had 4293 patients with 480 deaths (11.2%) observed mortality (10.25-12.16), predicted (10.93-11.54). Overall, there were 698 deaths in 14,799 patients (4.7%), observed mortality (4.37-5.06), predicted (4.72-4.95). CONCLUSION: EuroSCORE is a simple, objective and up-to-date system for assessing heart surgery, soundly based on one of the largest, most complete and accurate databases in European cardiac surgical history. We recommend its widespread use. PMID- 10456396 TI - Surgical management of transposition of great arteries associated with multiple ventricular septal defects. AB - OBJECTIVE: The presence of associated multiple ventricular septal defects (VSDs) increases the risk of the anatomic repair for transposition of the great arteries (TGA). The aim of this study was to define the optimal management of this complex anomaly. METHODS: Between January 1988 and December 1998, 45 patients underwent anatomic repair of TGA associated with multiple VSDs. The median age was 50 days and the median weight 4 kg. Eighteen (40%) had undergone previous palliation including 17 pulmonary artery banding procedure (PAB), seven associated with coarctation repair and one isolated coarctation repair. The perimembraneous septum was involved in 24 patients, the trabecular in 43, the inlet in seven and the infundibular in two. Closure of the VSDs included Dacron or pericardial patchs and matress sutures. The initial approach was through right atriotomy which was sufficient in 15 patients. VSDs were closed through right ventriculotomy in 13 patients, through pulmonary artery in six, through the aorta in one and in the remaining (n = 10) combined approaches were used. Only one patient required left apical ventriculotomy. RESULTS: There were five hospital deaths (11%; 70% CL: 6-18%) including the one early reoperation for residual VSD closure. Five patients had successful early reoperation for secondary PAB for residual VSD. Three late deaths occurred (7%; 70% CL: 3-13%). At the last visit, 95% of survivors were asymptomatic and without any cardiac medication. CONCLUSION: Mid-term survival with good quality of life can be achieved following either one or two-stage repair of this complex anomaly. In the presence of VSD closure failure a secondary PAB may be the procedure of choice. PMID- 10456397 TI - Median sternotomy single stage complete unifocalization for pulmonary atresia, major aorto-pulmonary collateral arteries and VSD-early experience. AB - OBJECTIVE: It is a prospective study to assess the results of median sternotomy, single stage complete unifocalization and repair for ventricular septal defect (VSD), pulmonary atresia and major aorto pulmonary collateral arteries (MAPCAs). METHODS: From June 97 to August 98, 20 patients were treated with single stage complete unifocalization and repair. Their ages ranged from 6 months to 11 years. Through median sternotomy, all MAPCAs were dissected and looped. On cardiopulmonary bypass, MAPCAs were anastomosed to native pulmonary arteries (PAs) or to MAPCAs. VSD was closed if possible and RV to PA continuity was established with a homograft conduit. If complete repair was not suitable, central shunt was done from ascending aorta to reconstructed PA with a polytetrafluroethylene graft. The patients were divided into three groups according to the arborization pattern in the lungs. Group 1 had well formed native PAs with MAPCAs, group 2 had hypoplastic PAs with MAPCAs and group 3 had only MAPCAs. RESULTS: Twenty patients had 21 procedures. All MAPCAs were unifocalized with tissue-to-tissue anastomosis for future growth, except one in whom polytetrafluroethylene tube graft was used to attain the confluence. In group 1, all seven patients had complete unifocalization and repair. In group 2, four patients had RV to PA conduit and two patients had central shunt. In group 3, three patients had complete repair, three patients had RV to PA conduit and one patient had central shunt. There were three deaths, two in group 2 and one in group 3. The first patient died due to a wrong decision to close the VSD, the second patient died due to missed large MAPCA in preoperative angio and the third patient was a 7-year-old boy who died with irreversible pulmonary vascular changes due to unprotected MAPCAs. CONCLUSIONS: To conclude, complete repair/RV PA conduit/central shunt should be done according to the size of the total pulmonary vasculature in patients with group 1, 2 and 3 with protected PAs/MAPCAs and in hypoplastic or absent PAs with unprotected MAPCAs (less than 1 year) and protected MAPCAs. We are yet to determine the surgical procedure to be performed in hypoplastic/absent PAs with unprotected MAPCAs more than 1 year. It is very essential to delineate all the MAPCAs up to the level of the diaphragm preoperatively. PMID- 10456398 TI - Ventricular outflow tracts after Kawashima intraventricular rerouting for double outlet right ventricle with subpulmonary ventricular septal defect. AB - OBJECTIVE: To determine whether or not the ventricular outflow tracts can be efficiently constructed in patients with double outlet right ventricle with subpulmonary ventricular septal defect by the Kawashima intraventricular rerouting in which the morphologically right ventricular outlet is divided into two, one for the systemic and the other for the pulmonary circulations. METHODS: The intraventricular rerouting procedure was carried out in nine patients with this particular malformation. Age at repair ranged from 35 days to 3 years old. The distance between the attachments of the tricuspid and the pulmonary valves was 10 mm or greater in all except one patient in whom the measured value was 3 mm. Resecting subaortic musculature appropriately, a tailored patch, either oval shaped (in seven) or heart-shaped (in two), was placed to construct an unobstructed channel for the left ventricular outflow tract with its diameter greater than that of the anticipated normal aortic orifice at the time of repair. For an unobstructed channel to the pulmonary arteries, enlargement of the right ventricular outflow tract was carried out using a patch in six. RESULTS: All patients survived the operative procedure. On postoperative catheterization, mean pulmonary arterial pressure was 15 +/- 8 mmHg, and cardiac index was calculated as 3.3 +/- 0.6 l/min per m2. It proved that the constructed left ventricular outflow tract can become larger in the longer term. Pressure gradient across the left ventricular outflow tract was greater than 20 mmHg in two patients in the intermediate term. One of these two underwent reoperation for the obstruction 10 years after the initial repair. It was suspected that use of a heart-shaped internal conduit, which seems to result from inadequate conal resection, was one of the possible causes of such obstruction in the longer term. Pressure gradient of 47 mmHg was seen across the right ventricular outflow tract in one patient, although this patient has undergone no reoperation. Enlargement of the right ventricular outflow tract could minimize postoperative obstruction for the pulmonary pathway. CONCLUSIONS: The intraventricular rerouting remains one of the attractive surgical options for repair in this particular setting, in terms of successful construction of the ventricular outflow tracts. PMID- 10456399 TI - Cytokine and S100B levels in paediatric patients undergoing corrective cardiac surgery with or without total circulatory arrest. AB - OBJECTIVES: Neurological damage following cardiopulmonary bypass (CPB) is difficult to objectively evaluate in infants. In adults, serum elevations of astroglial S100B correlate with proven brain injury independent of operative temperature. The deleterious effects of inflammatory cytokines, generated during CPB, on the brain have not been studied in infants using S100B as a marker for cerebral injury. METHODS: Twelve neonates, weighing 3.3 +/- 0.2 kg (total circulatory arrest group (TCA)) and 12 infants weighing 7.0 +/- 1.0 kg (cardiopulmonary bypass group (CPB)) underwent corrective cardiac surgery for various pathologies. Serial blood samples on induction, at the end of CPB, 30 min, 2 h and 24 h after the administration of protamine, were taken. The resultant plasma was frozen to -80 degrees C and stored for batch analysis. Cytokines were measured using ELISAs and S100B using a luminometric assay. RESULTS: The TCA group were younger and experienced a longer perfusion time than the CPB group (137 +/- 8 vs. 113 +/- 7, P = 0.04). The mean TCA time was 23 +/- 4 min. The TCA group had significantly higher levels of IL-6 (P = 0.001), IL-8 (P = 0.005) and S100B (P = 0.002) at 24 h. C5b-9 levels were significantly lower in the TCA group: end of CPB (P = 0.001), 30 min (P < 0.001), 2 h (P = 0.002). There was a weak, but significant correlation between IL-6 levels at the end of CPB and S100B levels 2 h later (r = 0.55, P = 0.03). Long extubation times were associated with high 24-h S100B levels (r = 0.52, P = 0.01). CONCLUSIONS: (1) The TCA group have prolonged rises of IL-6, IL-8 and S100B. (2) The TCA group generates significantly lower complement. (3) Astroglial injury, seen after surgery, may, in part, be cytokine mediated. PMID- 10456400 TI - Angioscopic video-assisted pulmonary endarterectomy for post-embolic pulmonary hypertension. AB - OBJECTIVES: To assess whether the use of video-assisted angioscopy would increase the outcome of pulmonary thromboendarterectomy (PTE). METHODS: PTE included a median sternotomy, intrapericardial dissection of the superior vena cava, institution of cardiopulmonary bypass, deep hypothermia and sequential circulatory arrest periods. It was always performed through two separate arteriotomies on both main intrapericardial pulmonary arteries, into which a rigid 5 mm angioscope connected to a video camera was introduced to increase the visibility and endarterectomies. RESULTS: From January 1996 to July 1998, 68 consecutive patients (35 males and 33 females) aged 54.3 +/- 13.5 years underwent PTE. Patients were in New York Heart Association (NYHA) class II (n = 2), III (n = 43) or IV (n = 23) with the following hemodynamics: mean pulmonary arterial pressure (PAP) 54 +/- 13 mmHg; cardiac output (CO): 3.8 +/- 0.8 l/min, and total pulmonary resistance (TPR): 1207 +/- 416 dyne x s x cm(-5). The cumulated circulatory arrest time was 23 +/- 12 min and postoperative length of ventilatory support 10 +/- 12 days. Nine patients died, for an overall in-hospital mortality of 13.2%. The functional outcome in surviving patients was significantly improved (P < 0.0001) both clinically (NYHA class 3.2 +/- 0.5 vs. 1.3 +/- 0.6) and hemodynamically (PAP (mmHg) 53.1 +/- 13 vs. 30.2 +/- 11.8, CI (l/min per m2) 2.1 +/- 0.5 vs. 2.8 +/- 0.6, TPR (dyne x s x cm(-5)) 1174 +/- 416 vs. 519 +/- 250). CONCLUSIONS: Video-assisted angioscopy improves the quality and degree of pulmonary endarterectomy expanding the indications to include patients with previously inaccessible distal disease. PMID- 10456401 TI - Thoracic outlet syndrome: influence of personal history and surgical technique on long-term results. AB - OBJECTIVE: Long-term results after surgery for thoracic outlet syndrome (TOS) are reviewed in terms of personal histories and surgical techniques. METHODS: Forty eight operations were performed in 37 patients. In 21 instances, the picture was one of ordinary TOS, in eight TOS was traumatic and in nine the picture was sub acute. Cervical ribs were excised through a supraclavicular approach (in seven cases), and first ribs through transthoracic, transaxillary or supraclavicular approaches (in 25, 15 or one, respectively). Long-term follow-up was obtained in 41 cases and averaged 11.7 years. RESULTS: Surgical decompression was successful in 28 cases (68%), including all patients with traumatic TOS (8/8) and seven with sub-acute symptoms (7/9). Outcome was good in five of seven supraclavicular cervical rib resections, and in 23 of 34 first rib excisions. First rib resections performed transaxillary had shorter post-operative stays, fewer complications. CONCLUSION: Surgical decompression is more successful when TOS is traumatic or sub-acute. When involved, a cervical rib can be resected through a supraclavicular approach, since the procedure is easy and has little morbidity. The transaxillary approach should be preferred for first rib resections because of shorter post-operative stays and fewer complications than after the transthoracic approach. PMID- 10456402 TI - Video-assisted thoracoscopic surgery does not deteriorate postoperative pulmonary gas exchange in spontaneous pneumothorax patients. AB - OBJECTIVES: Video-assisted thoracoscopic surgery (VATS) is generally recognized as a less invasive method than thoracotomy. However, the influence of VATS on postoperative pulmonary gas exchange has yet to be evaluated. METHODS: Thirty eight patients with spontaneous pneumothorax were randomized into bullectomy by VATS (n = 20) or axillary thoracotomy (n = 18). Gas exchange was assessed by using hot wire mass spectrometer, and blood gas analysis preoperatively and postoperatively at 1, 3, 6, 12, 24, and 48 h and on days 4 and 6. Post operative pain control was managed by continuous epidural morphine injection and administration of diclofenac sodium orally or suppository. Postoperative atelectasis was assessed by daily chest roentgenograms. RESULTS: VATS patients had continuously higher PaO2 than axillary thoracotomy at 12, 48 h and day 4 postoperatively (P < 0.05). Alveolar-arterial oxygen tension gradient in VATS patients was significantly less than that in patients with axillary thoracotomy from the 6th h throughout to the 4th day postoperatively (P < 0.01). Use of postoperative analgesics and the incidence of peripheral atelectasis were more frequent in patients with axillary thoracotomy. CONCLUSIONS: Bullectomy via VATS was less deleterious to pulmonary gas exchange. Axillary thoracotomy caused worsening of gas exchange postoperatively due to incisional pain, chest wall deformity, and peripheral atelectasis. PMID- 10456403 TI - Complications in the native lung after single lung transplantation. AB - OBJECTIVES: Single lung transplantation is a viable option for patients with end stage pulmonary disease; despite encouraging results, we observed serious complications arising in the native lung. We retrospectively reviewed 36 single lung transplants to evaluate the incidence of complications arising in the native lung, their treatment and outcome. METHODS: Between 1991 and 1997, 35 patients received 36 single lung transplants for emphysema (16), pulmonary fibrosis (14), lymphangioleiomyomatosis (4), primary pulmonary hypertension (1) and bronchiolitis obliterans (1). The clinical records were reviewed and the complications related to the native lung were divided into early (up to 6 weeks after the transplant) and late complications. RESULTS: Nineteen complications occurred in 18 patients (50%), leading to death in nine (25%). Early complications (within 6 weeks from the transplant) were bacterial pneumonia (1), overinflation (3), retention of secretions with bronchial obstruction and atelectasis (1), hemothorax (1), pneumothorax (1) and invasive aspergillosis (3); one patient showed active tuberculosis at the time of transplantation. Two patients developed bacterial pneumonia and invasive aspergillosis leading to sepsis and death. The other complications were treated with separate lung ventilation (1), bronchoscopic clearance (1), chest tube drainage (1) and wedge resection and pleurodesis (mechanical) by VATS (1). One patient with hyperinflation of the native lung eventually required pneumonectomy and died of sepsis. The patient with active tuberculosis is alive and well after 9 months of medical treatment. Late complications were recurrent pneumothorax (4), progressive overinflation with functional deterioration (2), aspergillosis (1) and pulmonary nocardiosis (1). Recurrent pneumothorax was treated with chest tube drainage alone (1), thoracoscopic wedge resection and/or pleurodesis (2) and pneumonectomy (1); hyperinflation was treated with thoracoscopic lung volume reduction in both cases; both patients with late infectious complications died. CONCLUSIONS: After single lung transplantation, the native lung can be the source of serious problems. Early and late infectious complications generally result in a fatal outcome; the other complications can be successfully treated in most cases, even if surgery is required. PMID- 10456404 TI - Reproducibility of thoracic aortic diameter measurement using computed tomographic scans. AB - OBJECTIVES: Decisions to recommend elective surgical repair of thoracic aortic aneurysms (TAA) may be based on size or expansion rate, which are used as indices of the risk of rupture. Measurement error may thus affect clinical decision making. In order to evaluate the reproducibility of aortic diameter measurements of TAA, we assessed departmental inter- and intra-observer variability of measurement of pre-selected computed tomographic scan images of aneurysmal segments of the thoracic aorta. METHODS: We compared measurements of minimum aortic diameter made by four observers in 50 pre-selected scans and at different times by two observers using a calliper method and a measurement tool within the scan. Differences in measured dimension were analysed using Wilcoxon's signed ranks test and the repeatability assessed using the method of Bland and Altman. RESULTS: There were no significant inter-observer differences among three observers but there were significant differences between another observer and two other observers (P < 0.05). No significant intra-observer differences existed. The best intra-observer repeatability was 2.25 while the worst inter-observer repeatability was 4.37. The mean and maximum difference in measurement were +/ 0.88 mm and +/-8.0 mm, respectively. Variability of measurement increased with aortic diameter. CONCLUSIONS: Calliper measurement of TAA is an acceptable measurement method for surveillance of TAA but appears most accurate with a single observer. Increasing error is seen with increasing diameter which may compound error in estimation of expansion rate. Standardisation of technique is advisable for multiple observers and aortic units should adopt quality assurance protocols to minimise error. PMID- 10456405 TI - The effect of chronic mechanical circulatory support on neuroendocrine activation in patients with end-stage heart failure. AB - OBJECTIVE: To evaluate if the improvement of patients supported with a Novacor was associated with a normalization in neuroendocrine activity. METHODS: Six patients had a Novacor implanted for end-stage heart failure. Four patients were transplanted after a mean of 4.5 months (range 3-6). One patient was weaned after 5 months and one died of a cerebral haemorrhage 5 weeks after implantation. Analysis of neuroendocrine activity was made prior to implantation and after 14, 30, 60 and 90 days. Levels of aldosterone, renin, cortisol, testosterone and T3 were measured using radio-immunoassays. Twenty-four hour urinary collections were made for assessment of adrenaline and noradrenaline excretion. RESULTS: Renin activity fell to normal after 14 days (16 +/- 3.0 ng/ml per h to 4.28 +/- 2.1 ng/ml per h, P < 0.05) and was maintained at 90 days. A similar picture was seen with aldosterone (1.5 +/- 0.4 nM to 0.12 +/- 0.07 nM, P < 0.05). Norepinephrine (67.46 +/- 14.1 microg/24 h) and epinephrine 12.9 +/- 2.5 microg/24 h) fell to normal physiological levels during the same time period. Cortisol levels were above normal pre-implantation but fell by day 30 (665.25 +/- 80.0 nM to 461.8 +/- 43.0 nM, P < 0.01). T3 and testosterone were lower than normal pre-implantation (T3 50 +/- 9.5 ng/dl vs. 90-200 ng/dl, testosterone 6.83 +/- 1.7 nM vs. 13-35 nM). T3 normalized after 90 days (81 +/- 11.7 ng/dl) and testosterone after 60 days (16.3 +/- 1.7 nM). CONCLUSION: Neuroendocrine function is abnormal in patients with cardiac failure who require circulatory support. The Novacor improved this, but metabolic recovery was delayed. The positive effect on the neuroendocrine axis, in the absence of activation of other endocrine systems, suggests that prolonged support may be well tolerated. PMID- 10456406 TI - Aortic valve replacement in the octogenarians: perioperative outcome and clinical follow-up. AB - OBJECTIVES: To determine long-term results of aortic valve replacement (AVR) in patients 80 years old or older, and assess the factors influencing perioperative outcome. METHODS: Data were reviewed on 83 consecutive octogenarians, undergoing aortic valve replacement between 1992 and 1997. There were 66 women and 17 men (mean age: 82.8 years). Fifty-seven patients (69%) were in New York Heart Association (NYHA) class III-IV and six had previous myocardial infarction. Three patients had previous percutaneous aortic valvuloplasty. There were 19 urgent procedures (23%). Coronary artery bypass grafting (CABG) was performed on 21 patients (25%). Possible influence of preoperative and operative variables on early and late mortality was performed with univariate and multivariate statistical analysis, and survival was estimated with the Kaplan-Meier method. RESULTS: Operative mortality was 13% (9% for AVR, 24% for AVR-CABG). Postoperative complications were respiratory failure in 19 patients, atrial fibrillation in 19, hemodialysis in four, myocardial infarction in four and stroke in two patients. Five patients required pacemaker insertion for permanent atrioventricular block. Median hospital stay and intensive care unit stay were 19.8 +/- 12.2 days and 7.9 +/- 3.4 days, respectively. Multivariate predictors of hospital death (P < 0.05) were percutaneous aortic valvuloplasty, NYHA class IV, and urgent procedure. Mean follow-up was 26.5 months. Survival at 1, 2, and 5 years was 98.5 +/- 1.4% (63 patients at risk), 93.4 +/- 3.2% (47 patients at risk), and 78.2 +/- 6.9% (six patients at risk), respectively. Preoperative myocardial infarction and urgent procedure were independent predictors of late death. At most recent follow-up, 91% were angina free and 81% were in class I-II. CONCLUSIONS: Aortic valve replacement in octogenarians can be performed with acceptable mortality. These results stress the importance of early operation on elderly patients with aortic valve disease. Both long-term survival and functional recovery are excellent. PMID- 10456407 TI - Reoperations for acute prosthetic thrombosis and pannus: an assessment of rates, relationship and risk. AB - OBJECTIVE: Mechanical valvular prostheses have the advantage of longevity but carry a risk of thrombosis which is dependant on valve design, materials and host related interface. While pannus is common to both biologic and mechanical valves, acute prosthetic thrombosis is mostly a complication of mechanical valves; therefore we investigated to find rates and risk of these obstructive complications. METHODS: Between 1/1/70 and 31/12/97, 2680 patients received at least one mechanical prosthesis in the aortic or mitral or tricuspid position and a total of 3014 operations were performed. Follow-up included 18523 years and was 98% complete. Incidence rates, Kaplan-Meier estimates, modeling of the hazard and multivariate analysis in the hazard domain were used in the analysis. RESULTS: Overall survival was 76%, 64%, 51%, 38.5% and 29% at 5, 10, 15, 20 and 25 years, respectively. It was significantly better in aortic than in mitral than in double prosthesis. 290 patients received a single reoperation, 37 a second, six a third and one a fourth reoperation. Two-hundred and fifty-one of these reoperations were exclusively due to malfunction of mechanical prosthesis, nine to malfunction of both mechanic and biologic prostheses. Most frequent reoperative indications was dehiscence (133), pannus (48) and thrombosis (29). The linearized rate of reoperations for pannus was 0.24%/patient per year, for valvular thrombosis 0.15%/patient per year. The shape of the thrombotic hazard was constant (at random) and the relative risk 12 times higher for tricuspid prosthesis, seven times higher for mitral prosthesis. Multivariate analysis controlling for prosthetic position, age, sex and prosthetic size, showed a 67% risk reduction with larger prosthesis (>27 mm), a 69% risk reduction with the Sorin tilting disk prosthesis and an 83% risk reduction with the bileaflet prosthesis. Pannus hazard shows a delayed exponential rise and was two times higher in tricuspid and three times higher in mitral position. Multivariate analysis showed a 50% risk reduction with larger prosthesis, an 11 times higher hazard of old (caged-disk, caged ball) prosthesis and a three times higher hazard of Lillehei-Kaster prosthesis. Reoperation for thrombosis has a 62% perioperative (30 days) survival compared to 92% survival of pannus reoperation. CONCLUSIONS: Mechanical valves have a low incidence of reoperation, mostly for prosthetic dehiscence. Pannus development is the next frequent complication increasing with time since implant, therefore in this series it was related to old valvular models and tilting disk prosthesis, with longer follow-up. Acute thrombosis occurs significantly earlier than pannus formation. Despite shorter follow-up we are therefore confident that bileaflet prostheses are less prone to this complication and pannus is a rare early etiologic factor. Thrombosis has very high operative risk as compared to pannus, justifying the present trend to thrombolysate selected cases. PMID- 10456408 TI - Long term results of mitral valve repair: posterior papillary muscle repositioning versus chordal shortening. AB - OBJECTIVE: Mitral valve repair is considered as the gold standard to treat mitral regurgitation. However anterior leaflet prolapse in the posterior paramedial and paracommissural area remains a challenging problem. Indeed several elongated chordae may arise from a single posterior papillary muscle head which does not allow safe separate chordal shortening (CS). We therefore suggest use of papillary muscle repositioning in such cases. METHODS: In a cohort of 180 mitral valve repair performed between 1989 and May 1998, we have retrospectively studied 100 consecutive patients who underwent anterior leaflet repair in the posterior paramedial and paracommissural area. Group I (n = 60) had posterior papillary muscle repositioning (PPMR) and group II (n = 40) had CS. There was no statistical difference between the two groups concerning age, functional class and left ventricular function. Etiology was similar in both groups, degenerative process being predominant. At echocardiogram, regurgitation was graded 3.4/4 in both groups. There was no statistical difference concerning preoperative ejection fraction, end systolic and end diastolic left ventricular diameter. RESULTS: There were no in-hospital deaths in group I and two deaths in group II not related to mitral valve repair. Mean follow up is 26.4 +/- 24.2 months in group I and 46.1 +/- 28.8 months in group II. No patient was lost to follow up. Severe mitral regurgitation was not observed. Mean regurgitation at follow up was 0.8 +/ 0.7 in group I and 0.8 +/- 0.8 in group II (P = n.s.); there was no statistical difference between the two groups concerning postoperative ejection fraction, end systolic and end diastolic left ventricular diameter. There was no late cardiac death in either group and there were no thromboembolic events. Actuarial survival rate is 100% and 94.4% in group I and 92% and 84.4% in group II at 2 and 6 years, respectively. CONCLUSION: This experience shows that PPMR provides as good longterm results as CS to repair anterior leaflet prolapse in posterior paramedial and paracommissural area with lesser morbidity and mortality. PMID- 10456409 TI - Neural network pattern recognition analysis of graft flow characteristics improves intra-operative anastomotic error detection in minimally invasive CABG. AB - OBJECTIVE: The intra-operative assessment of the quality of anastomosis in minimally invasive coronary artery bypass surgery (CABG) is critical. Recent investigations demonstrated that flow probes used intra-operatively to assess anastomotic errors may give the surgeon a false sense of confidence as only severely stenotic anastomoses (>90%) could be reliably detected. We developed a neural network system using graft flow data and assessed its potential to improve anastomotic error detection. METHODS: Mammary to LAD grafts (n = 46) were constructed in mongrel dogs off-pump. Continuous beat-to-beat graft flow was recorded using transit-time flow probes. Various degrees of anastomotic stenoses (0-100%) were created by an additional suture. The degree of anastomotic stenosis was confirmed by postoperative angiography. A learning vector quantization neural network was created using heart rate, mean aortic pressure, mean systolic, maximum systolic, minimum systolic, mean diastolic, maximum diastolic, minimum diastolic, and mean graft flows. In addition, a spectral analysis of the flow waveforms was performed and the magnitude and phase of the first five harmonics were used to further develop the neural network. RESULTS: The neural network pattern recognition system was 94% accurate in detecting any stenosis >50%. To validate the model, a testing set was used with 20% of the data values, and the accuracy remained at 100% above chance alone. CONCLUSION: Pattern recognition of transit-time flow probe tracings using neural network systems can detect anastomotic errors significantly better than the surgeon's visual assessment, thereby improving the clinical outcome of minimally invasive CABG. PMID- 10456410 TI - Thoracodorsal artery as a free graft for coronary artery bypass grafting. AB - Arterial myocardial revascularization using different arterial conduits as mammary, radial, gastroepiploic, subscapular and epigastric arteries are well documented. This report describes a preparation and use of thoracodorsal artery as a free graft for coronary artery bypass grafting. The preparation and removal of thoracodorsal artery were performed through right axilla. The artery was used as a free conduit for left anterior descending artery (LAD) in a 58-year-old female for the second revascularization. The saphenous veins, mammary arteries, right radial artery and epigastric artery were not available. The perioperative course was uneventful and we can recommend the graft as an alternative for such cases. PMID- 10456411 TI - Subpulmonary obstruction in transposition of the great arteries due to aneurysm of the membranous ventricular septum. PMID- 10456412 TI - Slide tracheoplasty for congenital funnelshaped tracheal stenosis (a 9-year follow-up of the first case) PMID- 10456413 TI - Improving the assessment of the economic impact of parasitic diseases and of their control in production animals. AB - This paper reviews the ways in which the economic impact of parasitic diseases of production animals have been evaluated. It then discusses the shortfalls of such studies, as well as the opportunities for improving the quality of economic impact assessments and their value to decision makers in the future. The paper first identifies the impacts that are specific to parasitic diseases. It then goes on to review the abundant literature on estimating the total costs of diseases. The authors argue that this approach severely limits the opportunity for economic assessments to aid decisions in disease control and research. The paper then reviews the literature on studies of avoidable costs, before discussing ways in which economic impact assessments can be enhanced. These issues include greater emphasis on incorporating the lost productivity potential caused by parasitic diseases, greater emphasis on valuing actual rather than intended control measures, and greater emphasis on quantifying the productivity effects at the societal level, particularly in the developing world. PMID- 10456414 TI - Integrated and biological control of parasites in organic and conventional production systems. AB - Organic and other non-intensive animal production systems are of growing importance in several countries worldwide. In contrast to conventional farms, parasite control on organic farms is affected by several of the prescribed changes in management e.g. access to the outdoors in the summer and in most countries, a ban on preventive medication, including use of anti-parasiticides. Organic animal production relies heavily on grazing, and pasture or soil related parasites are thus of major importance. Several studies in northern temperate climate have indicated that outdoor production of pigs, primarily sows, and laying hens results in heavier and more prevalent helminth infections compared to conventional intensive production under indoor conditions. In organic dairy cattle, parasitic gastroenteritis in heifers may be more prevalent. In a short to medium term perspective, integrated control may combine grazing management with biological control using nematophagous micro-fungi, selected crops like tanniferous plants and on conventional farms, limited use of anti-parasiticides. At present, the non-chemotherapeutic control of pasture related infections is based mainly on grazing management strategies. Preventive strategies, where young, previously unexposed stock, are turned out on parasite-free pastures, can be used for grazing first season dairy heifers and in all-in-all-out poultry production. Evasive strategies aim at avoiding disease producing infections of a contaminated area by moving to a clean area and may be relevant for ruminants and pigs. In cattle, effective control of nematodes can be achieved by repeated moves of the herd or alternate grazing with other species. High stocking rates seem to be an important risk factor. In pig production, the effect of paddock rotation on parasite infections is largely unknown and studies are warranted. Control of nematodes by larvae-trapping fungi, or perhaps in the future by egg-destroying fungi, looks promising for ruminants and certain monogastric animals but delivery systems and practical dosing regimes integrated with grazing management have to be developed. In conclusion, good prospects are expected for acceptable parasite control without a heavy reliance on anti-parasiticides through integration of the above mentioned procedures but future studies are needed to confirm their efficacy under practical farming conditions. PMID- 10456415 TI - Nutrition-parasite interaction. AB - The interactions between host nutrition and parasitism in ruminants are viewed within a framework that accounts for the allocation of scarce nutrient resources, such as energy and protein, between the various competing body functions of the host. These include functions that are the direct result of parasitism. Since it is proposed that the host gives priority to the reversal of the pathophysiological consequences of parasitism over other body functions, it is to be expected that improved nutrition will always lead to improved resilience. On the other hand, it is proposed that the function of growth, pregnancy and lactation are prioritised over the expression of immunity. Thus, improved nutrition may affect the degree of expression of immunity during these phases. The framework is useful at highlighting areas of future research on host/parasite/nutrition interactions. Its suggestions can account for the observations of the periparturient relaxation of immunity in reproducing females, as well as the reduction in worm burden in small ruminants supplemented with additional protein. Although developed for gastrointestinal nematodes in ruminants, the concepts of the framework should be applicable to the interactions of nutrition in other parasitic diseases. PMID- 10456416 TI - Parasites, biodiversity, and population dynamics in an ecosystem in the high arctic. AB - The abomasa of 163 Svalbard reindeers (Rangifer tarandus platyrhynchus) from Nordenskiold Land, Spitsbergen (78 degrees N, 15 degrees E) were examined for adult and juvenile nematodes. Spitsbergen has midnight sun from late April to late August, arctic night from late October to mid-February, permafrost, and snow cover from October to June. Plant growth is restricted to 6-8 weeks, usually starting mid-June. In the reindeer calves, which are born in June prevalence and intensity of infection increased over the winter. All young (1 year) and adult (older than 1 year) reindeers were infected. Variations in worm burden, fraction of adult nematodes, male to female ratio in the nematodes, number of adult nematodes, number of juvenile nematodes, and distribution of juvenile nematodes on length groups were analysed for relationships with sex and age of host, and time of the year. The material was further analysed for relationships between the reindeer's content of stored fat and worm burden and its elements. It is concluded that transmission of nematodes to the reindeer continues over the winter and that arrested larval development has not been strongly selected for in this High Arctic locality. Infection continued to increase in adult males but not in adult females with age. This is explained as a balance between gain and loss of worms in the females connected to levelling off of food intake at end of somatic growth. In adult male reindeers there was a negative trend between stored fat and infection at all seasons. In reproducing females there were significant negative relationships in winter-spring, while in yeld females no significant relationship was found. The nematodes belonged to the following taxa: Ostertagia gruehneri m. gruehneri, O. gruehneri m. arcticus, Marshallagia marshalli m. marshalli, M. marshalli. m. occidentalis, Teladorsagia circumcincta m. circumcincta, T. circumcincta. m. trifurcata, and T. circumcincta. m. davtiani. There appears not to be any specially adapted species among the helminth parasites of reindeer (Rangifer tarandus) and muskoxen (Ovibos moschatus) in the High Arctic. In the Low Arctic Uminmakstrongylus pallikuukensis of muskoxen may be an example of such adaptation. PMID- 10456417 TI - PCR-based technology in veterinary parasitology. AB - DNA technology is having a major impact in many areas of veterinary parasitology. In particular, the polymerase chain reaction (PCR) has found broad applicability because its sensitivity permits enzymatic amplification of gene fragments from minute quantities of nucleic acids derived from limited amounts of parasite material. This paper discusses some recent applications of PCR-based methods to parasites and highlights their usefulness or potential for those of veterinary importance. The focus is on PCR tools for the accurate identification of parasites and their genetic characterisation, the diagnosis of infections, the isolation and characterisation of expressed genes, the detection of anthelmintic resistance, and mutation scanning approaches for the high resolution analysis of PCR products. PMID- 10456418 TI - Immunology of helminth infections, with special reference to immunopathology. AB - Immune responses resulting in immunity to infection or disease, share the same basic humoral and cellular mechanisms. While immunity to helminth infection has evolved to mediate rapid elimination of the parasite, the strategies evolved by the parasites themselves aim to delay this rejection process and ensure the survival and distribution of their progeny. Ineffective or incomplete immunity results in persistence of parasites or their products within the host tissues, inappropriate or chronic stimulation by parasite antigens, hyper-reactivity and tissue damage or immunopathology. A long standing classification by Gell and Coombs identifies four major types of hypersensitivity responses accounting for most of the immunopathogenesis, three of which are mediated by antibody and one, delayed type hypersensitivity (DTH), by T cells. This paper aims to give a short review of these four classical hypersensitivity reactions with particular reference to infections of large animals with helminth parasites. In addition, in view of the functionally different helper T cell subsets now identified, the existing DTH response is redefined as DTH Type 1 (Th-1 mediated) and two new classes of T cell-dependent DTH responses are proposed; DTH Type II, associated with the Th-2 type cytokines IL-4 and IL-5 and eosinophilic granuloma formation, and DTH Type III, associated with IL-4 and TGF-beta and fibrosis. Finally, some implications of immunopathology on parasite control strategies are discussed. PMID- 10456419 TI - Frontiers in anthelmintic pharmacology. AB - Research in anthelmintic pharmacology faces a grim future. The parent field of veterinary parasitology has seemingly been devalued by governments, universities and the animal industry in general. Primarily due to the success of the macrocyclic lactone anthelmintics in cattle, problems caused by helminth infections are widely perceived to be unimportant. The market for anthelmintics in other host species that are plagued by resistance, such as sheep and horses, is thought to be too small to sustain a discovery program in the animal health pharmaceutical industry. These attitudes are both alarming and foolish. The recent history of resistance to antibiotics provides more than adequate warning that complacency about the continued efficacy of any class of drugs for the chemotherapy of an infectious disease is folly. Parasitology remains a dominant feature of veterinary medicine and of the animal health industry. Investment into research on the basic and clinical pharmacology of anthelmintics is essential to ensure chemotherapeutic control of these organisms into the 21st century. In this article, we propose a set of questions that should receive priority for research funding in order to bring this field into the modern era. While the specific questions are open for revision, we believe that organized support of a prioritized list of research objectives could stimulate a renaissance in research in veterinary helminthology. To accept the status quo is to surrender. PMID- 10456421 TI - Parasites in cultured and feral fish. AB - Parasites, causing little apparent damage in feral fish populations, may become causative agents of diseases of great importance in farmed fish, leading to pathological changes, decrease of fitness or reduction of the market value of fish. Despite considerable progress in fish parasitology in the last decades, major gaps still exist in the knowledge of taxonomy, biology, epizootiology and control of fish parasites, including such 'evergreens' as the ciliate Ichthyophthirius multifiliis, a causative agent of white spot disease, or proliferative kidney disease (PKD), one of the most economically damaging diseases in the rainbow trout industry which causative agent remain enigmatic. Besides long-recognized parasites, other potentially severe pathogens have appeared quite recently such as amphizoic amoebae, causative agents of amoebic gill disease (AGD), the monogenean Gyrodactylus salaris which has destroyed salmon populations in Norway, or sea lice, in particular Lepeophtheirus salmonis that endanger marine salmonids in some areas. Recent spreading of some parasites throughout the world (e.g. the cestode Bothriocephalus acheilognathi) has been facilitated through insufficient veterinary control during import of fish. Control of many important parasitic diseases is still far from being satisfactory and further research is needed. Use of chemotherapy has limitations and new effective, but environmentally safe drugs should be developed. A very promising area of future research seems to be studies on immunity in parasitic infections, use of molecular technology in diagnostics and development of new vaccines against the most pathogenic parasites. PMID- 10456420 TI - Chemo- and thermosensory neurons: structure and function in animal parasitic nematodes. AB - Nematode parasites of warm-blooded hosts use chemical and thermal signals in host finding and in the subsequent resumption of development. The free-living nematode Caenorhabditis elegans is a useful model for investigating the chemo- and thermosensory neurons of such parasites, because the functions of its amphidial neurons are well known from laser microbeam ablation studies. The neurons found in the amphidial channel detect aqueous chemoattractants and repellants; the wing cells-flattened amphidial neurons-detect volatile odorants. The finger cells digitiform amphidial neurons-are the primary thermoreceptors. Two neuron classes, named ADF and ASI, control entry into the environmentally resistant resting and dispersal dauer larval stage, while the paired ASJ neurons control exit from this stage. Skin-penetrating nematode parasites, i.e. the dog hookworm Ancylostoma caninum, and the threadworm, Strongyloides stercoralis, use thermal and chemical signals for host-finding, while the passively ingested sheep stomach worm, Haemonchus contortus, uses environmental signals to position itself for ingestion. Amphidial neurons presumably recognize these signals. In all species, resumption of development, on entering a host, is probably triggered by host signals also perceived by amphidial neurons. In the amphids of the A. caninum infective larva, there are wing- and finger-cell neurons, as well as neurons ending in cilia-like dendritic processes, some of which presumably recognize a sequence of signals that stimulate these larvae to attach to suitable hosts. The functions of these neurons can be postulated, based on the known functions of their homologs in C. elegans. The threadworm, S. stercoralis, has a complex life cycle. After leaving the host, soil-dwelling larvae may develop either to infective larvae (the life-stage equivalent of dauer larvae) or to free-living adults. As with the dauer larva of C. elegans, two neuron classes control this developmental switch. Amphidial neurons control chemotaxis to a skin extract, and a highly modified amphidial neuron, the lamellar cell, appears to be the primary thermoreceptor, in addition to having chemosensory function. The stomach worm, Haemonchus contortus, depends on ingestion by a grazing host. Once ingested, the infective larva is exposed to profound environmental changes in the rumen. These changes stimulate resumption of development in this species. We hypothesize that resumption of development is under the control of the ASJ neuronal pair. Identification of the neurons that control the infective process could provide the basis for entirely new approaches to parasite control involving interference with development at the time and place of initial host-contact. PMID- 10456422 TI - Important parasites in poultry production systems. AB - Poultry now accounts for 30% of all meat consumed. Parasites are a problem where ever poultry are raised, whether in large commercial operations or in small back yard flocks, and economic losses can be significant. This paper will briefly review the major protozoan, helminth, and arthropod species in poultry including pathogenesis. Other topics will include the importance of the interaction of other diseases and parasites, and control of the infection by chemotherapy, management, and immunity. PMID- 10456423 TI - Recent advances in Neospora and neosporosis. AB - Neospora caninum has emerged as a major cause of abortion in cattle in many countries. This paper reviews recent advances in the life cycle and biology of Neospora with the emphasis on bovine neosporosis. The role of the recently discovered oocyst stage of N. caninum in the epidemiology of neosporosis is discussed. Progress made in serologic diagnosis of N. caninum infection is discussed. There is no vaccine for preventing Neospora-induced abortions in cattle or to prevent oocyst shedding in dogs. PMID- 10456424 TI - Neuropeptide Y has a stimulatory action on feeding behavior in goldfish (Carassius auratus). AB - The purpose of the present study was to elucidate the possible role of neuropeptide Y (NPY) in the feeding regulation in fish. We examined the effects of intracerebroventricular (i.c.v.) or intraperitoneal (i.p.) neuropeptide Y administration on food intake in satiated goldfish, at different time intervals postinjection (0-2, 2-8 and 0-8 h). Food intake was significantly increased by i.c.v. administered neuropeptide Y (1 microg) at 2 h postinjection, while no significant differences in food intake were observed after i.p. treatment. The neuropeptide Y receptor antagonist, neuropeptide Y-(27-36), totally counteracted the stimulatory action of neuropeptide Y on feeding. The possible involvement of neuropeptide Y in the eating behavior evoked by food deprivation has been investigated. Food deprivation by either 24 or 72 h significantly increased feeding, and the neuropeptide Y receptor antagonist attenuated such feeding stimulation. From our findings, we suggest, first, that neuropeptide Y is involved in feeding central regulation in goldfish, acting via specific neuropeptide Y receptors, and second, that hypothalamic neuropeptide Y would be released in response to food deprivation, contributing to generate the consequent eating behavior stimulation in Carassius auratus. PMID- 10456425 TI - Neuroprotection by metabotropic glutamate receptor glutamate receptor agonists: LY354740, LY379268 and LY389795. AB - In rat cortical neuronal cultures, metabotropic glutamate (mGlu) receptor agonists: LY354740 (+)-2-aminobicyclo[3.1.0]hexane-2,6dicarboxylate); LY379268 ( )-2-oxa-4-aminobicyclo[3.1.0]hexane-4,6-dicarboxylate, and LY389795 (-)-2-thia-4 aminobicyclo[3.1.0]-hexane-4,6-dicarboxylate, were neuroprotective against toxicity induced by N-methyl-D-aspartic acid (NMDA), kainic acid and staurosporine as measured by release of lactate dehydrogenase (LDH) activity into culture supernatants and DNA fragmentation by oligonucleosome formation. The potencies of the agonists were at least 100 times greater in reducing nucleosome formation than LDH release indicating a differential effect on neurons dying by apoptosis than by necrosis. In vivo studies showed that LY354740 was able to mediate a partial protection against apoptosis in CA1 hippocampal cells under ischaemic conditions where substantial CA1 cell loss occurred. The effects of the agonists in vitro were: (a) reversed by mGlu receptor antagonist LY341495, (b) enhanced by the presence of glial cells, (c) abrogated by RNA and protein synthesis inhibitors, and (d) unaltered by inhibition of endogenous adenosine activity. These results suggest that group II mGlu receptor agonists may represent a novel therapeutic strategy for the treatment of neurodegenerative diseases. PMID- 10456426 TI - Distinct features of seizures induced by cocaine and amphetamine analogs. AB - Seizure-related emergencies caused by stimulants of abuse have been increasing. To better understand the nature of these drug-induced convulsions, we characterized the seizure patterns associated with high doses of cocaine, and the amphetamine analogs, methamphetamine, methylenedioxymethamphetamine (MDMA) and 4 methylaminorex. The features of the stimulant-induced seizures were distinct and included the following: (1) the duration of convulsive activity was shortest for cocaine and longest for methamphetamine, (2) only MDMA produced a secondary clonic phase after the initial ictal event, and (3) 4-methylaminorex manifested a very steep dose-response curve. Differential preventive profiles of anticonvulsant agents on the stimulant-induced seizures also were observed. For example, cocaine-related seizures were most effectively prevented by, while methamphetamine-induced seizures were completely refractory to, phenytoin pretreatment. The only anticonvulsants which appeared to influence methamphetamine-related convulsions were diazepam and valproate. A unique feature of 4-methylaminorex was that related seizures were almost completely blocked by the calcium channel blocker, flunarizine. PMID- 10456427 TI - Effect of coadministration of caffeine and either adenosine agonists or cyclic nucleotides on ketorolac analgesia. AB - Caffeine potentiation of ketorolac-induced antinociception in the pain-induced functional impairment model in rats was assessed. Caffeine alone was ineffective, but increased the effect of ketorolac without affecting its pharmacokinetics. Intra-articular administration of adenosine and N6-cyclohexyladenosine (CHA, an adenosine A1 receptor agonist), but not 2-p-(2-carboxyethyl)phenethylamino-5'-N ethylcarboxamidoadenosine hydrochloride (CGS-21680, an adenosine A2A receptor agonist), significantly increased ketorolac antinociception. This effect was not local, as contralateral administration was also effective. Ipsilateral and contralateral administration of adenosine and CHA also increased antinociception by ketorolac-caffeine. Intra-articular 8-Bromo-adenosine cyclic 3',5'-hydrogen phosphate sodium or 8-Bromo-guanosine-3',5'-cyclophosphate sodium (cGMP) given ipsilaterally or contralaterally did not affect ketorolac-induced antinociception. Nevertheless, ipsilateral, but not contralateral, administration of 8-Br-cGMP significantly increased antinociception by ketorolac-caffeine, suggesting a local effect. The results suggest that caffeine potentiation of ketorolac antinociception is mediated, at least partially, by a local increase in cGMP and rule out the participation of adenosine receptor blockade. PMID- 10456428 TI - Differential effects of nitric oxide synthase inhibitor, 7-nitroindazole, on discriminative stimulus and somatic effects of naloxone in morphine-dependent rats. AB - Our previous report suggested that antagonists acting at NMDA receptors attenuate discriminative stimulus effects of naloxone in morphine dependent rats. Nitric oxide (NO) is a putative second messenger which mediates NMDA receptor activation. The present study evaluated behavioral effects of NO synthase inhibitor, 7-nitroindazole in morphine-dependent rats trained to discriminate 0.1 mg/kg naloxone from saline. 7-Nitroindazole did not significantly affect naloxone's discriminative stimulus effects but decreased naloxone-induced weight loss and abolished expression of several withdrawal signs--diarrhea, scream on touch, tremor and 'wet dog'-like shaking suggesting different mechanisms for subjective and somatic components of opioid withdrawal. PMID- 10456429 TI - Pindolol increases extracellular 5-HT while inhibiting serotonergic neuronal activity. AB - The effects of pindolol, a beta-adrenoceptor blocker/putative 5-hydroxytryptamine (5-HT)1A/1B antagonist, on both the single-unit activity of serotonergic neurons in the dorsal raphe nucleus (DRN) and extracellular 5-HT levels in the caudate nucleus, were examined in freely moving cats. Administration of (+)-pindolol (1 and 10 mg/kg, s.c.) decreased neuronal activity and increased 5-HT levels in a dose- and time-dependent manner. The subsequent administration of WAY-100635 [N [2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cycloh exanecarboxamide] (0.2 mg/kg, s.c.), a selective 5-HT1A receptor antagonist, blocked pindolol-induced neuronal suppression and potentiated 5-HT output. These results indicate that pindolol may be acting at the level of the nerve terminal to increase 5-HT. PMID- 10456430 TI - Increased nitroglycerin-induced relaxation by genistein in rat aortic rings. AB - The effect of genistein, a tyrosine kinase inhibitor, on nitroglycerin-induced relaxation was examined in rat aortic rings contracted by phenylephrine. In rat aortic rings, genistein (10(-5) M and 3x10(-5) M), a tyrosine kinase inhibitor, but not daidzein, an analogue of genistein, increased relaxation induced by nitroglycerin in a concentration-dependent manner. Iberiotoxin, an inhibitor of Ca2+ -activated K+ channels, inhibited the relaxation induced by nitroglycerin, but it did not affect the effect of genistein. Glibenclamide, an inhibitor of ATP sensitive K+ channels, did not affect the relaxation induced by nitroglycerin. Theophylline, an inhibitor of cyclic AMP-dependent phosphodiesterase, increased the relaxation induced by nitroglycerin, and genistein (10(-5) M) failed to affect the relaxation induced by nitroglycerin in the presence of theophylline. Genistein also inhibited the activity of cyclic AMP-dependent phosphodiesterase. In addition, 6-[4-(4'-pyridyl)amino phenyl]-4,5-dihydro-3(2H)-pyridazinone hydrochloride, an inhibitor of cyclic GMP-inhibitable cyclic AMP phosphodiesterase, inhibited the relaxation induced by nitroglycerin. These results suggest that, in the rat aortic rings, genistein inhibits cyclic AMP dependent phosphodiesterase activities, resulting in the increase of the relaxation induced by nitroglycerin. PMID- 10456431 TI - Tight coupling between the rate of rise of Ca2+ transient and peak twitch contraction in guinea-pig papillary muscle. AB - We evaluated the relationship between cytoplasmic Ca2+ concentration ([Ca2+]i) and force in guinea-pig papillary muscles loaded with a fluorescent Ca2+ indicator, fura-PE3. In the absence of ryanodine, [Ca2+]i transient and force were altered by changing extracellular Ca2+ concentration and stimulation frequency, and also by adding methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4-(2 trifluoromethylphenyl)-pyri dine-5-carboxylate (Bay K 8644) or ouabain. Under these conditions, the peak force correlated linearly with the maximal rate of rise of [Ca2+]i (gamma = 0.948) more than the peak [Ca2+]i transient (gamma = 0.737). Ryanodine inhibited the increase in the maximal rate of rise of [Ca2+]i resulting in abolishment of the correlation between force and the maximal rate of rise of [Ca2+]i. These results suggest that the maximal rate of rise of [Ca2+]i reflects Ca2+ release from the sarcoplasmic reticulum, and this fraction of [Ca2+]i is crucial for determining the amplitude of twitch contractions when the sarcoplasmic reticulum is intact in guinea-pig papillary muscle. PMID- 10456432 TI - Low dose of lipopolysaccharide induces a delayed enhanced nitric oxide-mediated relaxation in rat aorta. AB - Delayed effect on vascular reactivity of isolated aorta was studied after injection of a single low dose of lipopolysaccharide (0.5 mg/kg i.p.). The maximal vascular effect was observed 72 h after lipopolysaccharide administration with an increase in maximal endothelium-dependent relaxing response to acetylcholine and parallelly a decrease in contractile response to phenylephrine. The change in contractile response was nullified by endothelium removal as well by in vitro aortic rings incubation with N(omega)-monomethyl-L-arginine but not with indomethacin. A low dose of lipopolysaccharide induces a delayed enhanced nitric oxide-mediated vascular relaxation which could contribute to its delayed anti-ischemic properties in ischemic tolerance phenomenon. PMID- 10456433 TI - Contractile effect of (+)-glaucine in the isolated guinea-pig ileum. AB - The intestinal effects of (+)-glaucine [(S)-1,2,9,10-tetramethoxyaporphine] were studied using the guinea-pig ileum. (+)-Glaucine (10-300 microM) induced ileal contractions. The contraction was not affected by tetrodotoxin, atropine, hexamethonium, propranolol, naloxone, methysergide, N(G)-nitro-L-arginine methyl ester, SR141716A (a cannabinoid CB1 receptor antagonist) or SR140333 (a tackykinin NK1 receptor antagonist) plus SR48968 (a tackykinin NK2 antagonist). (+)-Glaucine-induced contraction was reduced by indomethacin, nordihydroguaiaretic acid or bisindolylmaleimide I and abolished by verapamil and nifedipine. These results suggest that (+)-glaucine-induced contraction involves activation of voltage-dependent Ca2+ channels and protein kinase C and could be mediated by the release of arachidonic acid metabolites. PMID- 10456434 TI - Role of endogenous nitric oxide in the nitric oxide donor-induced plasma extravasation of mouse skin. AB - The role of endogenous nitric oxide (NO) and prostanoids in the increase in microvascular permeability induced by NO donors was investigated in the mouse skin by a dye leakage method. Subcutaneous (s.c.) injection of 1-hydroxy-2-oxo-3 (3-aminopropyl)-3-isopropyl-1-triazene (NOC 5), 1-hydroxy-2-oxo-3,3-bis(2 aminoethyl)-1-triazene (NOC 18) and sodium nitroprusside dose-dependently increased local dye leakage. While indomethacin inhibited the dye leakage elicited by these NO donors, N(G)-nitro-L-arginine methyl ester (L-NAME) inhibited the effect of NOC 5 and NOC 18 but not of sodium nitroprusside. These results suggest that endogenous NO, in addition to the prostanoid biosynthesis, is involved in the dermal microvascular permeability increase induced by the NOC series NO donors. PMID- 10456435 TI - Nucleoside diphosphate kinase associated with membranes modulates mu-opioid receptor-mediated [35S]GTPgammaS binding and agonist binding to mu-opioid receptor. AB - The role of nucleoside diphosphate kinase (NDKP), which converts GDP to GTP, in the coupling of mu-opioid receptors to G protein was investigated in membranes of Chinese hamster ovary cells stably transfected with the cloned rat mu-opioid receptor (rmor). Endogenous NDPK activity in membranes was determined to be 0.60+/-0.02 micromol/mg protein/30 min UDP (at 10 mM), a competitive substrate of NDPK for GDP with no effect on guanine nucleotide binding to G proteins, reduced basal [35S]GTPgammaS binding and unmasked morphine-stimulated [35S]GTPgammaS binding to pertussis toxin-sensitive G proteins, indicating that [35S]GTPgammaS binding to NDPK accounts for part of its high basal binding. UDP increased the extent of morphine-induced increase in [35S]GTPgammaS binding in the presence of GDP, most likely by reducing basal binding and inhibiting conversion of GDP to GTP. ATP greatly reduced morphine-induced increase in [35S]GTPgammaS binding, whereas AMP-PCP (adenylyl-(beta,gamma-methylene)-diphosphoate tetralithium salt), which cannot serve as the phosphate donor for NDPK, did not, demonstrating that effects of ATP is mediated by the NDPK product GTP. In addition, GDP and ATP increased the Kd and lowered the Bmax of the agonist [3H]DAMGO ([D-Ala2,N-Me Phe4,Gly5ol]-Enkephalin) for the mu-opioid receptor and GDP alone increased Kd, most likely through their conversion to GTP by NDPK. Addition of exogenous NDPK enhanced the inhibitory effects of GDP and combined GDP and ATP on [3H]DAMGO binding. Thus, NDPK appears to play a role in modulating signal transduction of and agonist binding to mu-opioid receptors. PMID- 10456436 TI - The profiles of human and primate [3H]Nalpha-methylhistamine binding differ from that of rodents. AB - Characterization of the histamine H3 receptor in rodent species has been extensive but limited characterization has been done with primate or human tissue. We have characterized the binding of [3H]Nalpha-methylhistamine to cynomolgus monkey and human brain membranes to determine whether there are any significant differences among species' pharmacology. In monkey, [3H]Nalpha methylhistamine bound, in a guanine nucleotide-sensitive fashion, to an apparently homogeneous class of sites at equilibrium (K(D) = 1.4 nM, Bmax = 34 fmol/mg protein). The profile of binding was broadly similar to that of rodents, with a couple of significant differences. Most notably, the potency of the histamine H3-receptor-specific antagonist thioperamide (Ki = 240 nM) was substantially less than reported for rodents and under assay conditions that yield a two-site curve fit in rodents only a single class of thioperamide binding sites was detected in monkey. Burimamide, however, yielded a two-site curve fit (KiH = 6.7 nM, KiL = 1100 nM) independent of the presence of sodium in the assay, as it does in rodents. Characterization of the human brain histamine H3 receptor showed that it was similar to the monkey and not rodent receptor. Our findings indicate that differences between primate and rodent histamine H3 receptors of potentially serious importance for the discovery of antagonists active in humans do exist. PMID- 10456438 TI - Is it time for a reassessment of prognostic features in B-cell chronic lymphocytic leukemia? AB - Prognostic assessment of B-cell chronic lymphocytic leukemia (CLL) patients is generally based on either Rai or Binet clinical staging systems. However, new biological parameters which reflect the clinical heterogeneity of disease are under investigation. Cellular and molecular features including tumor cell proliferation, immunophenotype, adhesion molecules expression and release, karyotypic abnormalities and biological findings of increased angiogenesis have been correlated with tumor mass and survival. It is not clear, however, whether the newly identified prognostic parameters will eventually replace clinical variables representative of tumor mass. More likely, biological parameters might be incorporated into clinico-prognostic models thus leading to the formulation of a clinico-biological system for CLL. PMID- 10456437 TI - Metformin modulates insulin receptor signaling in normal and cholesterol-treated human hepatoma cells (HepG2). AB - The effects of the biguanide anti-hyperglycemic agent, metformin (N,N'-dimethyl biguanide), on insulin signaling was studied in a human hepatoma cell line (HepG2). Cells were cultured in the absence (control cells) or in the presence of 100 microM of a cholesterol derivative, hemisuccinate of cholesterol. Cholesterol hemisuccinate-treatment alters cholesterol and lipid content of HepG2 and modulates membrane fluidity. Cholesterol hemisuccinate-treatment induces a decrease in insulin responsiveness and creates an 'insulin-resistant' state in these cells. Exposure to 100 microM of metformin resulted in a significant enhancement of insulin-stimulated lipogenesis in control and cholesterol hemisuccinate-treated cells. In control cells, metformin altered glycogenesis in a biphasic manner. In cholesterol hemisuccinate-treated cells, metformin inhibited basal glycogenesis but restored insulin-stimulated glycogenesis. Hence, to understand the mechanism of metformin action, we analyzed early steps in the insulin signaling pathway, including insulin receptor autophosphorylation, mitogen-activated-protein kinase and phosphatidylinositol 3-kinase activities, in both control and cholesterol hemisuccinate-treated cells. Overall, the results suggest that metformin may interact with the insulin receptor and/or a component involved in the early steps of insulin signal transduction. PMID- 10456439 TI - Gamma-irradiation-induced DNA single- and double-strand breaks and their repair in chronic lymphocytic leukemia cells of variable radiosensitivity. AB - Gamma-irradiation-induced DNA single- and double-strand break (SSB and DSB) formation and their repair kinetics in normal hematopoietic cells and in leukemic lymphocytes was investigated using alkaline and neutral comet assays. The cells were isolated by density gradient centrifugation from peripheral blood of patients with chronic lymphocytic leukemia (CLL) and from healthy study subjects. Furthermore, CD34+ progenitor cells isolated with immunomagnetic beads from bone marrow of non-leukemic persons were investigated. The cytotoxicity of 137Cs irradiation was determined in vitro in peripheral blood mononuclear lymphocytes from 36 CLL patients and from 8 healthy donors using radioactive leucine incorporation assay in 4-day culture. A dose-dependent increase in DNA migration was observed in alkaline (SSBs) and neutral (DSBs) gel electrophoresis when the cells were exposed to gamma-irradiation doses up to 10.4 Gy. After irradiation with doses of 2.4 and 5.4 Gy, the cells repaired their single- and double-strand breaks almost completely. The formation and repair of DNA strand breaks were essentially similar in all normal cell populations investigated and in CLL cells. The gamma-irradiation-induced cytotoxicity did not correlate with DNA strand break formation and repair capacity. According to these results, the differences of gamma-irradiation tolerance among individual CLL cases and among healthy persons are explicable in terms other than DNA strand break formation or repair. PMID- 10456441 TI - The GEN.S: a fortuitous finding of a routine screening test for hereditary spherocytosis. AB - As part of the evaluation of the GEN.S (Coulter), we compared the Mean Corpuscular Volume (MCV) to the Mean Spherized Corpuscular Volume (MSCV) assessed during the reticulocyte count procedure under hypo-osmotic conditions. A sub group of patients with hereditary spherocytosis (HS) was singled out: in all of them, the MSCV became smaller than the MCV. As the cell volume normally increases in red cells derived from other patients in the same conditions, we decided to further study the reason for this particular behaviour of HS red cells. Whereas normal red cells are able to undergo an osmotic expansion, the spherocytes reach a critical osmotic volume leading to cell fragmentation consistent with the decrease of MSCV. This fortuitous finding is likely to be a reliable improvement for the routine screening of HS. PMID- 10456440 TI - Ex vivo expansion CD34+/AC133+-selected autologous peripheral blood progenitor cells (PBPC) in high-risk breast cancer patients receiving intensive chemotherapy. AB - AC133 antibody provides an alternative to CD34 for the selection and characterization of cells necessary for engraftment in transplant situations. We studied the effect of stem cell factor (SCF), interleukin 3 (IL-3) and interleukin 11 (IL-11) on the ex vivo expansion of human CD34+/AC133+ progenitors isolated from leukapheresis products from chemotherapy plus granulocyte-colony stimulating factor (G-CSF) -mobilized adult donors. MiniMACS AC133+ isolated cells contained a mean of 85% (80-90) AC133+ cells. Enriched AC133+ cells co expressed CD34+ 80%, CD71low 36.6% and CD33+ 6.6%. After a seven-day ex vivo expansion in the presence of SCF + IL-3 + IL-11, the number of cells increased 19 times. These cells expressed a mean 12% CD34+ and 74% CD71+ (23% CD 71high) and 59% CD33+. This means that the absolute number of CD34+ cells increased slightly, the number of CD33+ increased 100 times and cells with high density CD71high (23%) appeared. These cells represent the cells committed to erythroid differentiation. The increase in the number of CFU-GM with various combinations of cytokines SCF + Il-3 + IL-11 will be discussed. The number of CFU-GM in culture after a seven-day ex vivo expansion in the presence of SCF + IL-3 + IL-11 increased 45 times. PMID- 10456442 TI - Autologous transplantation with peripheral blood stem cells in chronic lymphocytic leukemia. A phase III, randomized, multicenter study. GIMEMA. Gruppo Italiano per le Malattie Ematologiche dell'Adulto. GITMO-Gruppo Italiano Trapianto di Midollo Osseo. PMID- 10456443 TI - A randomized phase II trial of high-dose chlorambucil vs. fludarabine in patients with advanced B-chronic lymphocytic leukemia. EORTC Leukemia Cooperative Group. PMID- 10456444 TI - The 536C-->T transition in the human tissue factor pathway inhibitor (TFPI) gene is statistically associated with a higher risk for venous thrombosis. AB - Tissue factor pathway inhibitor (TFPI) is an important regulator in the extrinsic blood coagulation pathway. Although the regulatory biochemical role of TFPI is evident, the clinical significance of this proteinase inhibitor remains to be elucidated. The definition of a clinical TFPI deficiency seems to be more complex than that of other coagulation inhibitors because the activity and concentration of circulating TFPI can not be considered a true measure of in vivo levels. Its determination in plasma samples by immunological methods or functional assays has been shown to be inadequate in the detection of a clinical deficiency. Therefore, we screened genomic DNA samples of blood donors and thrombotic patients for alterations in the TFPI gene to assess the influence of a modified TFPI in venous thromboembolic diseases. We detected a single nucleotide substitution in exon 7 (536C-->T) leading to a proline to leucine exchange at amino acid position 151 of the protein ([P151L]TFPI) and found the prevalence of heterozygous carriers in German unrelated blood donors to be 0.2% (n = 5120). Four unrelated persons out of 14 probands carrying the genetic variation could be linked to venous thrombosis. For calculation of a potential risk for venous thrombosis for carriers of the mutation we investigated healthy blood donors about thrombotic events. 7 out of 308 blood donors were found to have a history of venous thrombosis, one of them carried the TFPI mutation. Statistical calculation showed a significant relative risk for venous thrombosis for individuals with the trait (odds ratio, 9.3; confidence interval, 1.8-48.6; p <0.01). PMID- 10456445 TI - Thrombophilic polymorphisms are common in women with fetal loss without apparent cause. AB - An association between fetal loss and thrombophilia has recently been described but has not been yet fully elucidated. We have evaluated prospectively the prevalence of the three common thrombophilic polymorphisms (TP) factor V G1691A (Leiden), thermolabile-methylenetetrahydrofolate reductase (TL-MTHFR) C677T and factor II G20210A mutations, in 76 women with fetal loss (> or =3 in first, > or =2 in second, > or =1 in third trimester) without apparent cause and 106 controls without fetal loss. Thirty seven out of 76 (49%) of the women in the fetal loss group had at least one TP compared to only 23/106 (22%) in the control group (p = 0.0001 ). Factor V-Leiden was more common in the fetal loss group 24/76 (32%) compared to the control group 11/106 (10%) (OR = 4.0, 95% CI: 1.8-8.8, p <0.001). Five of the 76 patients (7%) were homozygous for factor V-Leiden compared to none of the controls (p = 0.012). A trend, albeit no statistically significant difference was found between women with fetal loss and control groups regarding factor II G20210A (8% vs. 4% respectively, OR = 2.2, 95% CI: 0.6-8.0, p = 0.23) and MTHFR C677T (18% vs. 10% respectively, OR = 1.95, 95% CI: 0.83-4.6, p = 0.12). Combined TP were documented in 6/76 (8%) patients compared to 1/106 (1%) in controls (OR = 9.0, 95% CI: 1.1-76, p = 0.02). Second or third trimester fetal loss were more common cause of pregnancy termination in 37 patients with TP compared to 39 patients without TP (57/158 (36%) vs. 23/135 (17%) respectively, (p = 0.0004). Thrombophilic polymorphisms are common in women with fetal loss without apparent cause and are associated with late pregnancy wastage. Combinations of TP increase the risk for fetal loss. PMID- 10456446 TI - Lipoprotein(a) concentrations in women with a history of severe preeclampsia--a case control study. AB - A high concentration of lipoprotein(a) is associated with atherosclerotic disease. Atheroma may develop in spiral arteries in both preeclamptic and normal pregnancies, but they are much more common in preeclampsia, particularly in the decidual segments. We hypothesized that a high concentration of lipoprotein(a) is associated with the development of preeclampsia. We studied 40 women with a history of severe preeclampsia, 35 women with a history of preeclampsia and the (H)ELLP syndrome and 67 controls with a normal obstetric history. Lipoprotein(a) levels were measured at least 10 weeks post partum in the second half of a normal menstrual cycle. None of the women in the study or the control group were pregnant or used oral contraceptives. Lipoprotein(a) levels over the 90th percentile of the lipoprotein(a) distribution of our control group (420 mg/l) were defined as abnormal. There was a statistically significant higher prevalence of abnormal levels of lipoprotein(a) in women with a history of severe preeclampsia (33%) in comparison with women with a history of preeclampsia and (H)ELLP syndrome (9%) and with the control group (10%). We found that a history of severe preeclampsia and spontaneous abortion was associated with elevated lipoprotein(a) levels as a post-hoc finding. Whether spontaneous abortion and high levels of lipoprotein(a) are related remains to be demonstrated. PMID- 10456447 TI - Helicobacter pylori infection and the risk of myocardial infarction: role of fibrinogen and its genetic control. AB - The contribution of Helicobacter pylori (HP) infection to the risk of myocardial infarction was evaluated. The role of fibrinogen and its genetic control as a possible mechanism by which HP may influence myocardial infarction risk was explored in this context. A case-control study was performed in 101 patients with myocardial infarction and in 101 controls. HP infection was associated with an increased risk of myocardial infarction independently for confounding variables (OR 4.1, CI95: 1.8-9.4). HP infection was significantly associated with higher levels of fibrinogen, both in cases and in controls. Furthermore, there was an additive effect of HP infection and B2 allele of BclI fibrinogen polymorphism in increasing fibrinogen levels. HP infection showed a stronger effect on the risk of myocardial infarction in B2 allele carriers (OR 7.6, CI95: 1.8-31.6) as compared to subjects carrying the B1B1 genotype (OR 3.3, CI95: 1.2-9.2). We showed that a previous HP infection is a risk factor for myocardial infarction. An increase in fibrinogen levels is a possible mechanism by which HP may act. Concomitant conditions, like a genetic predisposition in increasing fibrinogen levels, seem to further increase the effect of HP on myocardial infarction risk. PMID- 10456448 TI - Genetic polymorphism of 5,10-MTHFR reductase gene in offspring of patients with myocardial infarction. AB - A family history of myocardial infarction is a major determinant of ischemic disease. A C->T677 polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene has been identified as a cause of mild hyperhomocysteinemia, a risk factor for arterial thrombosis. We have investigated the relationship between the MTHFR TT genotype and a family history of myocardial infarction in a cohort of 982 apparently healthy individuals. Subjects whose first-degree relatives suffered from a myocardial infarction, showed raised median age (p <0.001), total cholesterol (p <0.001) and plasma fibrinogen (p = 0.023) and a higher than normal frequency of C-reactive protein levels >0.33 mg/dl (p = 0.012). Moreover, when compared to subjects without such family history, a higher number of homozygotes for the T allele of the MTHFR gene (p = 0.027), and of the 4G allele of the plasminogen activator inhibitor-1 gene (p = 0.002) was found in the subsetting of the offspring of patients with myocardial infarction. In a multiple logistic regression analysis, age (OR 1.02 [95%-CI: 1.00-1.05]), total cholesterol (OR 1.40 [95%-CI: 1.14-1.71]), C-reactive protein levels >0.33 mg/l (OR: 1.87 [95% CI: 1.10-3.20]), plasminogen activator inhibitor-1 4G/4G (OR: 1.84 [95%-CI: 1.27 2.66]), and MTHFR TT genotype (OR 1.62 [95%-CI: 1.08-2.42]), were all associated with a family history of myocardial infarction. Thus, the MTHFR TT genotype independently accounts for the risk of a family history for myocardial infarction in the present setting. PMID- 10456449 TI - Characterization of native human thrombopoietin in the blood of normal individuals and of patients with haematologic disorders. AB - Thrombopoietin (TPO) isolated from thrombocytopenic plasma of various animal species has previously been shown to comprise only truncated forms of the molecule, presumably generated by proteolysis. Native TPO has now been partially purified from normal human plasma by immunoaffinity chromatography and was confirmed to be biologically active. Gel filtration in the presence of SDS revealed that TPO eluted in two peaks: a major peak corresponding to the elution position of fully glycosylated recombinant human TPO (rhTPO) consisting of 332 amino acid residues, and a minor peak corresponding to a smaller molecular size. Immunoblot analysis also revealed that most plasma-derived TPO migrated at the same position as fully glycosylated rhTPO, corresponding to a molecular size of approximately 80 to 100 kDa. Furthermore, the size distribution of circulating TPO in patients with various haematologic disorders did not differ markedly from that of plasma-derived TPO from healthy individuals. These results indicate that the truncation of circulating TPO is not related to disease pathophysiology, and that the predominant form of TPO in blood is a biologically active approximately 80- to 100-kDa species. The size distribution of TPO extracted from normal platelets was similar to that of TPO in plasma; the proportion of truncated TPO was decreased by prior incubation of platelets with hirudin. indicating that the endogenous truncated TPO, at least in platelet extract, was generated by thrombin mediated cleavage. PMID- 10456450 TI - Dermatan sulphate for the prevention of postoperative venous thromboembolism in patients with cancer. DOS (Dermatan sulphate in Oncologic Surgery) Study Group. AB - BACKGROUND: Patients undergoing surgery for cancer are at high risk for venous thromboembolism (VTE). Agents that selectively inhibit thrombin, such as dermatan sulphate, have potential for a favourable benefit-risk ratio in the prevention of this complication. METHODS: Patients scheduled for elective abdominal, thoracic, gynecologic or urologic surgery for cancer resection, were randomised to dermatan sulphate (600 mg intramuscularly on the second day before surgery, then 300 mg once daily), or calcium heparin (5,000 IU subcutaneously t.i.d., starting 2 hours before operation). Both treatments were continued until postoperative day 7 or until adequate mobilisation was achieved. Bilateral venography was scheduled at the end of treatment. Venograms were centrally assessed in blind conditions. The study outcomes were VTE (venographically proven deep vein thrombosis IDVT] or symptomatic, objectively confirmed pulmonary embolism) and bleeding complications. RESULTS: At 27 centres, 842 patients were randomised and underwent surgery (418 dermatan sulphate, 424 heparin). Efficacy was assessed in 521 patients with adequate venography and/or confirmed pulmonary embolism. DVT was observed in a total of 96 patients; symptomatic non-fatal pulmonary embolism developed in 2 patients (one per group), who also had DVT at venography. Postoperative VTE occurred in 40 of 267 patients on dermatan sulphate, 15.0%, versus 56 of 254 patients on heparin. 22.0% (p = 0.033). Relative risk reduction was 32.7% (95% confidence interval, 3.1 to 53.2%). The rate of bleeding complications in all operated patients was 6.9% with dermatan sulphate and 7.5% with heparin (confidence interval for the absolute risk difference, -4.1 to 2.9%). The inhospital mortality rate was 1.2% and 1.4%, respectively. CONCLUSIONS: In oncologic surgery, dermatan sulphate prevents VTE more effectively than heparin without increasing bleeding complications. PMID- 10456451 TI - Evaluation of the PFA-100 system in the diagnosis and therapeutic monitoring of patients with von Willebrand disease. AB - We have evaluated platelet function at high shear with the PFA-100 system in different subtypes of von Willebrand disease (vWD), before and after the intravenous infusions of desmopressin or a factor-VIII/von Willebrand factor (vWF) concentrate. Closure times with the PFA-100 system were determined for both the collagen/ADP and the collagen/epinephrine cartridges in 52 patients with vWD (9 type 1 "platelet normal", 5 type 1 "platelet-discordant", 8 type 1 "platelet low", 6 type 2A, 9 type 2B, 6 type 2M Vicenza. 6 type 3 and 3 acquired vWD) and 40 controls. Measurements were repeated 1 and 4 h after the i.v. infusion of desmopressin (0.3 microg/Kg) in 26 patients with types 1, type 2M Vicenza or type 2A vWD, or of a factorVIII/vWF concentrate (Alphanate HT, 60 U/Kg) in 4 patients with type 3 vWD. At all time points, vWF plasma levels and the bleeding time (Symplate II) were also determined. Baseline closure times were longer in vWD patients than in controls with both the collagen/ADP and the collagen/ epinephrine cartridges. The sensitivity of the PFA-100 system (88% and 87% with the two cartridges) was higher than that of the bleeding time (65%). Treatment with desmopressin normalized the closure times in patients with type 1 "platelet normal" or type 2M Vicenza vWD, had no significant effects in patients with type 1 "platelet-low", type 1 "platelet-discordant" or type 2A vWD. Infusion of a factorVIII/vWF concentrate in patients with type 3 vWD slightly shortened their prolonged closure times. In general, changes in PFA-100 were paralleled by shortenings of the bleeding times and increases in plasma vWF levels. The PFA-100 test reflects vWF-dependent platelet function under high shear stress and could be useful in the diagnosis and therapeutic monitoring of patients with vWD. PMID- 10456452 TI - Some factor VIII (FVIII) inhibitors recognise a FVIII epitope(s) that is present only on FVIII-vWF complexes. AB - A mild haemophilia A patient (LE) with an Arg2150His mutation in the C1 domain of the factor VIII (FVIII) light chain was shown to have anti-FVIII antibodies inhibiting wild type but not self FVIII. Polyclonal anti-FVIII antibodies of this patient were purified by affinity adsorption using recombinant FVIII (rFVIII) and/or plasma-derived FVIII-von Willebrand factor (vWF) complexes. A distinct population of antibodies was obtained that bound to FVIII-vWF complexes but not to rFVIII, indicating that an epitope was created by the association of FVIII to vWF. Such antibodies belonged to the IgG2 isotype, but the FVIII epitopes to which they bind could not be mapped with precision due to vWF dependency. Depletion experiments showed that anti-FVIII antibodies recognising FVIII-vWF complex also distinguished wildtype from mutated self FVIII, indicating that the Arg2150His mutation alters the B cell epitope formed by the association of FVIII to vWF. To determine whether the Arg2150His substitution also alters the formation of the FVIII-vWF complex, the interaction between mutated or normal FVIII with vWF was evaluated in plasma. The dissociation rate of mutated FVIII from vWF was found to be significantly increased. The presence of an Arg2150His mutation therefore results in the disappearance of a FVIII B cell epitope generated by the association of FVIII with vWF. Patients carrying such an Arg2150His mutation and receiving infusion of wild-type FVIII may therefore be at risk of developing inhibitors to allogeneic FVIII only. PMID- 10456453 TI - A collaborative study to establish the 2nd International Standard for Antithrombin Concentrate. AB - Eighteen laboratories participated in a collaborative study to calibrate a replacement for the 1st International Standard for Antithrombin. Concentrate (88/548). Excellent agreement between laboratories, as indicated by mean % gcv of 3.3 and 5.9 for functional and antigenic assays, was observed when the candidate concentrate (96/520) was assayed against the 1st International Standard for Antithrombin, Concentrate (88/548). The functional potency was found to be 7.9% (p<0.05) lower than the antigenic potency. Based on the results and with the agreement of the participants of this study and the authorisation of the Expert Committee on Biological Standardisation of the World Health Organisation, the antithrombin concentrate, coded 96/520, has been established as the 2nd International Standard for Antithrombin, Concentrate, with labelled potencies for both functional (4.7 IU/ampoule) and antigenic (5.1 IU/ampoule) activities. PMID- 10456454 TI - Antithrombogenic coating of stents using a biodegradable drug delivery technology. AB - To reduce the thrombogenic properties of coronary artery stents, a biodegradable polylactic acid (PLA) stent coating with an incorporated thrombin inhibitor and a platelet aggregation inhibitor has been developed. In an ex vivo human stasis model, its effect on platelets, plasmatic coagulation and its release characteristics were studied using whole blood. Bare steel and bare gold-surface stents were compared to steel and gold-surface stents coated with PLA (30 kDa) containing 5% polyethyleneglycol (PEG)-hirudin and 1% iloprost, with an empty tube as control. Markers of activated coagulation (prothrombin fragment F1-2 and thrombin-antithrombin III complex, TAT), were assayed and the release of drugs from the coating was assessed by aPTT and collagen-induced platelet aggregation. Bare steel and gold stents were completely covered by a blood clot, and high levels of coagulation markers (F1-2 fragment and TAT) were detected. No differences in the thrombogenic properties were found between bare gold or steel stents. Coated stents were free of blood clots and only minor elevations of markers were detected. Release data from in-vitro studies over 90 days showed a gradual release of the drugs with an initial exponential release characteristic for PEG-hirudin, slow release of iloprost and a 10% degradation of the PLA carrier. This drug releasing biodegradable coating effectively reduced thrombus formation independent of the metallic surface. PMID- 10456455 TI - Titanium is a highly thrombogenic biomaterial: possible implications for osteogenesis. AB - Titanium has superior osteointegrating properties compared to other biomaterials. The mechanism for this is unknown. During the initial phase of bone implantation the biomaterial comes into direct contact with whole blood. In this study we use a newly developed in vitro chamber model to compare different commonly used biomaterials in contact with whole blood. These materials were selected with respect to their different osteointegrating properties in order to correlate these properties with the response to whole blood. In the presence of 3 IU/ml of heparin only titanium induced macroscopic clotting. This was reflected by the generation of thrombin-antithrombin which was much increased in blood in contact with titanium compared with steel and PVC. The coagulation activation caused by titanium was triggered by the intrinsic pathway because the generation of FXIIa AT/C1 esterase inhibitor paralleled that of thrombin-antithrombin, and both thrombin-antithrombin complex and FXIIa-AT/C1 esterase inhibitor generation were abrogated by corn trypsin inhibitor, which is a specific inhibitor of FXIIa. The binding of platelets was increased on the titanium surface compared to the other biomaterial surfaces and the state of platelet activation was much more pronounced as reflected by the levels of beta-thromboglobulin and PDGF. This study indicates that titanium is unsuitable as a biomaterial in devices which are in direct contact with blood for a prolonged period. Furthermore, PDGF and other alpha-granule proteins e.g. TGF-beta, are known to be potent promotors of osteogenesis which suggests that the pronounced thrombogenic properties of titanium might contribute to the good osteointegrating properties. PMID- 10456456 TI - A novel splice acceptor site mutation which produces multiple splicing abnormalities resulting in protein S deficiency type I. AB - In an attempt to explore the molecular mechanisms for protein S deficiency, a patient with such a deficiency was examined at the DNA, RNA and protein levels. Nucleotide analyses revealed that the proband, the mother and the grandmother had a G-->C substitution in the invariant AG dinucleotide at the splicing acceptor site of intron A/exon 2. This patient was heterozygous for this substitution and the mutant allele was inherited from the proband's mother and grandmother. Reverse transcription-polymerase chain reaction analysis demonstrated several kinds of splicing abnormalities such as exon skipping and cryptic splicing, in addition to correct splicing. Semiquantitation of mRNA for the protein S gene revealed that the amount of the proband's mRNA was reduced to 60% of normal. Thus, this mutation impaired the normal processing of mRNA for the protein S gene, resulting in the subject's severe protein S deficiency. PMID- 10456457 TI - Involvement of Lys 62(217) and Lys 63(218) of human anticoagulant protein C in heparin stimulation of inhibition by the protein C inhibitor. AB - Inhibition of activated protein C (APC) by protein C inhibitor (PCI) is stimulated by heparin, whereas inhibition by alpha1-antitrypsin (AAT) is heparin independent. Three lysine residues located in a positively charged cluster in the serine protease domain of protein C (PC) were mutated to probe their involvement in the heparin stimulation of inhibition by PCI. These mutations were selected after analysis of the three-dimensional structure of APC and of molecular models for PCI and the APC-PCI complex. A double mutant, K62[217]N/K63[218]D, a single mutant, K86[241]S, and wild-type PC were expressed in embryonic human kidney 293 cells. Heparin stimulated the rate of inhibition of wt-APC by PCI approximately 400-fold, with second order rate constants (k2) in the absence and presence of heparin of 0.72 x 10(3) M(-1)s(-1) and 2.87 x 10(5) M(-1)s(-1), respectively. In contrast, heparin only yielded a 52-fold stimulation of the rate of inhibition of the double mutant APC by PCI as the rate constants in the absence and presence of heparin were k2 = 2.44 x 10(3) M(-1)s(-1) and k2 = 1.26 x 10(5)M(-1)s(-1), respectively. The double mutant K62N/K63D eluted at approximately 10% lower NaCl concentration from a heparin Sepharose column than the K86S mutant or wt-APC. These data suggest K62 and K63 in APC to be part of a heparin binding site which is important for heparin-mediated stimulation of inhibition of APC by PCI. PMID- 10456458 TI - Regulation of prothrombinase activity by protein S. AB - The independent effect of protein S as prothrombinase inhibitor has been proposed to depend on binding to both coagulation factors Va and factor Xa or on the binding to phospholipid thereby limiting the phospholipid available for prothrombinase activity. In this study we show that plasma concentrations of protein S (300 nM) equilibrated with the prothrombinase components (factor Va, factor Xa, phospholipid) cause a profound inhibition at low phospholipid concentrations (approximately 0.2 microM). This inhibition by protein S of prothrombinase activity is abrogated with increasing phospholipid concentrations. Modeling of the effect of protein S on prothrombinase based only on the reported affinity of protein S for phospholipids (Kd approximately 10(-8) M) in an equilibrium model (Clotspeed), predicted the experimentally obtained thrombin generation rates at low phospholipid in the presence of protein S based on the diminished available phospholipid binding sites for the prothrombinase components. Consistently, initial rates of prothrombinase activity are already maximally inhibited when protein S is preincubated with the phospholipid prior to the addition of factor Xa, factor Va and prothrombin. The results indicate that the order of addition of prothrombinase components and the availability of phospholipid may have a profound influence on observed effects of protein S on prothrombinase activity. All prothrombinase components (factor Xa, factor Va, phospholipid) become available during the course of the physiological thrombin generation. The effect of protein S was therefore studied on tissue factor induced, platelet-dependent thrombin generation. Protein S delayed and inhibited the rate of thrombin generation of tissue factor-induced thrombin formation when surface is provided at physiologic concentrations using isolated platelets (2 x 10(8)/ml). In contrast, protein S hardly affected thrombin generation in this model when platelets were pre-activated with collagen. Furthermore, the observed effects of addition of protein C and thrombomodulin in the absence or presence of protein S on tissue factor-induced, platelet-dependent thrombin generation, indicate that protein S and protein C may cooperate in the regulation of prothrombinase activity through independent mechanisms. PMID- 10456459 TI - Role of tissue factor in metastasis: functions of the cytoplasmic and extracellular domains of the molecule. AB - Tissue factor (TF) is a transmembrane glycoprotein that complexes with factor VIIa to initiate blood coagulation. It was reported in an earlier study that expression of high levels of TF in a human melanoma cell line promotes metastasis, and that the cytoplasmic domain of TF is required for this metastatic effect. To analyze the functions of the cytoplasmic and extracellular domains of TF in metastasis, two TF mutants were constructed; in one mutant alanine was substituted for each of the three serine residues in the cytoplasmic domain, preventing phosphorylation; in the other mutant alanine was substituted for four key residues in the extracellular domain, preventing binding of factor VIIa and consequently eliminating the initiation of blood coagulation by the TF-VIIa complex. Melanoma lines expressing high levels of either mutant form of TF were weakly metastatic in SCID mice, indicating that phosphorylation of the cytoplasmic domain and formation of a complex with VIIa by the extracellular domain are required for the full metastatic effect of TF. It was also found that increasing TF expression in human melanoma cells does not increase expression of vascular endothelial growth factor or promote growth and vascularization of tumors derived from the melanoma cells, suggesting that TF acts by a mechanism other than angiogenesis to promote metastasis. PMID- 10456460 TI - Adsorption of vitamin K-dependent proteins to live cell membranes measured under flow conditions. AB - Mechanisms mediating initial adsorption of coagulation proteins to live cells were investigated. Adsorption kinetics were examined under varying flow conditions using tracer-dilution techniques in perfused spherical monolayers of cells expressing tissue factor. At biologically relevant time and concentration ranges, rates exceeded by 2-12 fold the theoretical maximum calculated for steady state diffusion. Rates were correlated with aqueous-phase flux of reactants and were found to be largely independent of the density of reactive sites on the membrane. Average adsorption rate of factor VIIa at 4 etaM and flow velocity of 0.8 etam s(-1) was 5 x 10(7) s(-1) cm(-2). Adsorption rates of homologous coagulation factors IX and X under similar conditions were 5 and 9 x 10(7) s( 1)cm(-2). Results indicate that flow can effectively increase the rate of coagulation factor adsorption to the membrane of live cells. They also imply that factors affecting blood flow velocity and vessel permeability influence the rate of membrane-dependent coagulation reactions. PMID- 10456461 TI - Plasmin-alpha2-antiplasmin complex in patients with atrial fibrillation. Stroke Prevention in Atrial Fibrillation Investigators. AB - Plasmin-alpha2-antiplasmin complex (PAP) is an index of recent fibrinolytic activity. We examined PAP levels in patients with atrial fibrillation (AF) to determine whether these levels are correlated with clinical characteristics associated with stroke risk. We obtained blood for measurement of PAP in a non random sample of 586 patients with AF on entering the Stroke Prevention in Atrial Fibrillation III Study. PAP levels were measured with an ELISA assay. PAP values were transformed with a natural logarithm (PAPln) prior to all analyses. Older age, female gender, recent congestive heart failure, decreasing fractional shortening, recent onset of AF, and coronary artery disease were each univariately associated with higher levels of PAP (all p<0.05, two-sample t-test, simple linear regression). Older age, recent congestive heart failure, decreasing fractional shortening, and recent onset of AF were independently associated with higher PAP levels by multivariate analysis (linear regression). Among patients receiving warfarin, PAP levels were not correlated with INR levels (linear regression, p=0.60). Patients classified as high-risk for thromboembolism by our risk stratification criteria (systolic blood pressure > 160 mm Hg, prior thromboembolism, recent congestive heart failure, poor left ventricular function, and women over age 75) had higher PAP levels than low-risk patients (antilog mean PAPln 5.6 vs 4.9. p<0.001, two-sample t-test). PAP levels in patients with AF are associated with clinical characteristics predictive of thromboembolism. Elevated PAP levels are particularly associated with poor left ventricular function and are not affected by anticoagulation. PAP levels may be a marker of stroke risk in patients with AF. PMID- 10456462 TI - Relationship of plasminogen activator inhibitor-1 levels following thrombolytic therapy with rt-PA as compared to streptokinase and patency of infarct related coronary artery. AB - BACKGROUND: Type 1 plasminogen activator inhibitor (PAI-1) is considered to be risk factor for acute myocardial infarction (AMI). A rebound of circulating PAI-1 has been reported after rt-PA administration. We investigated the relationships between PAI-1 levels before and after thrombolytic therapy with streptokinase (SK) as compared to rt-PA and the patency of infarct-related arteries. METHODS AND RESULTS: Fifty five consecutive patients with acute MI were randomized to streptokinase or rt-PA. The plasma PAI-1 levels were studied before and serially within 24 h after thrombolytic administration. Vessel patency was assessed by an angiogram at 5+/-1days. The PAI-1 levels increased significantly with both rt-PA and SK as shown by the levels obtained from a control group of 10 patients treated with coronary angioplasty alone. However, the area under the PAI-1 curve was significantly higher with SK than with rt-PA (p<0.01) and the plasma PAI-1 levels peaked later with SK than with rt-PA (18 h versus 3 h respectively). Conversely to PAI-1 levels on admission, the PAI-1 levels after thrombolysis were related to vessel patency. Plasma PAI-1 levels 6 and 18 h after SK therapy and the area under the PAI-1 curve were significantly higher in patients with occluded arteries (p<0.002, p<0.04 and p<0.05 respectively). The same tendency was observed in the t-PA group without reaching significance. CONCLUSIONS: This study showed that the PAI-1 level increase is more pronounced after SK treatment than after t-PA treatment. There is a relationship between increased PAI-1 levels after thrombolytic therapy and poor patency. Therapeutic approaches aimed at quenching PAI-1 activity after thrombolysis might be of interest to improve the efficacy of thrombolytic therapy for acute myocardial infarction. PMID- 10456463 TI - A bispecific antifibrin-antiplatelet urokinase conjugate (BAAUC) induces enhanced clot lysis and inhibits platelet aggregation. AB - Thrombolysis is well established in the treatment of acute myocardial infarction. However, clinical application of thrombolytic agents has limitations with respect to efficacy and specificity. To achieve highly effective and at the same time clot-selective plasminogen activation urokinase was coupled to a bispecific antibody consisting of the monovalent Fab' from the antifibrin monoclonal antibody 59D8 and the monovalent Fab' from the anti-glycoprotein GPIIb/IIIa monoclonal antibody 7E3. The bispecific antifibrin-antiplatelet urokinase conjugate (BAAUC) was synthesized and characterized. Assays with either immobilized platelets, GPIIb/IIIa or fibrin showed an increase in plasminogen activation compared to uncoupled urokinase by 10-fold, 58-fold and 13-fold, respectivley (p < 0.0001 each). In vitro clot lysis was performed on platelet rich and fibrin-rich clots and revealed an up to 5-fold higher potency of BAAUC compared to uncoupled urokinase (p < 0.0001). In vitro platelet aggregation was effectively inhibited by the hybrid molecule, whereas urokinase had no effect. BAAUC and two monospecific urokinase-conjugates, UK-59D8-IgG and UK-7E3-(Fab')2 were compared with each other with regard to similar tests. In vitro clot assays with platelet-rich and platelet-poor clots were performed. BAAUC achieved a significantly higher plasminogen activation compared to each of the monospecific conjugates (p < 0.05, respectively). We conclude that BAAUC, a bispecific plasminogen activator with antifibrin and antiplatelet properties has the potency to lyse both fibrin-rich and platelet-rich thrombi with high efficacy and to effectively inhibit platelet aggregation. PMID- 10456464 TI - The 4G/5G polymorphism in the plasminogen activator inhibitor-1 gene is not associated with myocardial infarction. AB - Several studies have found an association between high plasminogen activator inhibitor-1 (PAI-1) levels and myocardial infarction. Whether this is causal or a consequence of atherosclerosis or tissue damage, remains unclear. Homozygous carriers of the 4G allele of the 4G/5G polymorphism in the PAI-1 gene have higher PAI-1 levels compared to carriers of the 5G allele in healthy persons in some studies, but not all. If PAI-1 levels are causally related to myocardial infarction, one would expect more homozygous carriers of the 4G allele among patients, provided that these carriers have high PAI-1 levels among healthy persons in that population. We investigated the distribution of this polymorphism in the "Study of Myocardial Infarctions Leiden" (SMILE), including 331 men with a myocardial infarction and 302 control subjects and measured PAI-1 antigen levels among the latter. Secondly, we looked into the association of cardiovascular risk factors with PAI-1 levels. We did not find an increase in risk of myocardial infarction in carriers of the 4G allele. Neither did we find an association, nor a trend, between the 4G/5G polymorphism and PAI-1 antigen levels in control subjects. Controls with obesity, hypertension, or who smoked had significant higher PAI-1 antigen levels compared with persons without these factors. High cholesterol and triglyceride levels were also associated with high PAI-1 antigen levels, and HDL-cholesterol levels showed an inverse association. We conclude that the 4G/5G polymorphism in the PAI-1 gene is not associated with the risk of myocardial infarction. As we did not find any association between this polymorphism and PAI-1 antigen levels in healthy persons, we cannot draw any conclusions about the causality of PAI-1 itself for myocardial infarction. PMID- 10456465 TI - Evidence that modifications of Lp(a) in vivo inhibit plasmin formation on fibrin- a study with individual plasmas presenting natural variations of Lp(a). AB - In the present study we have investigated the effect of individual variations in the concentration of Lp(a) on plasmin formation at the surface of fibrin. The plasma Lp(a) concentrations from 20 nephrotic children were high at flare-up of the disease (0.43+/-0.45 g/l) and decreased progressively with remission at both 6 weeks (0.28+/-0.24 g/l) and 6 months (0.24+/-0.288 g/l). In contrast, the concentration of plasminogen showed an inverse variation, with low values at flare-up (1.27+/-0.34 microM) and normal values at remission (1.66+/-0.17 microM at 6 weeks and 1.99+/-0.21 microM at 6 months). An increase in plasmin formation (from 0.62+/-0.49 to 0.73+/-0.61, and 0.84+/-0.75 pmol/well) and a decrease in apo(a) binding (from 5.45+/-2.42 to 4.54+/-2.12, and 3.93+/-1.51 fmol/well) on the surface of fibrin, were concomitantly observed from flare-up to remission at 6 weeks and at 6 months, respectively. Values for plasmin formation parallel the amount of plasminogen bound. The low concentration of plasminogen found at flare up may also have contributed to the increased binding of Lp(a) as indicated by a decrease in the maximal amount of Lp(a) bound (Bmax) to fibrin as a function of plasma plasminogen concentrations. Bmax was 1.51 fmol in the absence of plasminogen and decreased to 1.1 fmol and 0.93 fmol at respectively 1 and 2 microM of plasminogen. Altogether, these data constitute the first quantitative evidence indicating that plasmin formed at the surface of fibrin may vary with modifications of the concentration of Lp(a) in vivo. PMID- 10456466 TI - Persistent impairment of platelet aggregation following cessation of a short course dietary supplementation of moderate amounts of N-3 fatty acid ethyl esters. AB - The duration of the effect of a short-course (1-mo twice-daily) supplementation of moderate amounts (2.28 g) of n-3 fatty acid ethyl esters (FA) on platelet lipid composition and aggregation was compared with that of olive oil (3 g/d) supplementation in 14 healthy volunteers. The FA preparation employed contained eicosapentaenoic acid (EPA) and docosahexaenoic acids (DHA) in a ratio of 1:1.4. A marked rise (p <0.05) in the plasma and platelet content of EPA and DHA, and minimal changes in the content of arachidonic acid (AA) were documented at withdrawal of the n-3 FA supplementation. EPA/AA and DHA/AA ratios in platelet phospholipids showed that the FA accumulation persisted 8-12 wks after stopping the supplementation (p <0.05). The aggregation of platelets in response to collagen or ADP, and thromboxane B2 (TXB2) formation were impaired at withdrawal. The impaired aggregation lasted 8-12 weeks (p always <0.05), whereas TXB2 formation returned to basal values 4 weeks after stopping the n-3 supplementation. No correlation was found between impaired aggregation and TXB2 formation. In contrast, the impaired sensitivity to ADP (p = 0.036) and, to a lesser extent, to collagen (p = 0.068) were related to changes in the intracellular pH (pHi) of the Na+/H+ reverse transport. No changes in platelet composition or function were observed either during or following olive oil supplementation. These results document a long-lasting impairment of platelet sensitivity to ADP and collagen; changes in the pHi values of the Na+/H+ reverse transport, and a simultaneous persistent accumulation of EPA and DHA in platelet phospholipids, after stopping a short-course dietary supplementation of moderate amounts of n-3 fatty acid ethyl esters. PMID- 10456467 TI - New assay for measuring binding of platelet glycoprotein IIb/IIIa to unpurified von Willebrand factor. AB - Among the numerous variants of vWD, no patient with an abnormal vWF binding to GPIIb/IIIa has been described to date. To search for such potential variants, we developed a two-site assay for measuring the binding of purified GPIIb/IIIa to vWF in biological fluids and we used it to study a large series of plasmas from various types of von Willebrand disease (vWD) and recombinant vWF (rvWF). vWF in plasma or rvWF in culture medium was immobilized onto anti-vWF monoclonal antibodies (MoAb)-coated wells of microtiter plates. After incubation with either unlabeled GPIIb/IIIa and a 125I-anti-GPIIb/IIIa MoAb or 125I-GPIIb/IIIa, binding curves and binding isotherms were respectively established. Normal pool plasma and wild-type rvWF were used as reference samples. We tested plasmas from 85 normal subjects, 115 patients with different types of vWD (64 type 1, 2 type 3, 9 type 2A, 4 type 2M, 16 type 2B, 15 type 2N, 3 type IID and 2 acquired forms) and 50 patients with various bleeding disorders. Four mutated rvWF with 2A (Glu875Lys and Pro885Ser) or 2B (Dupl.Met540 and Val551Phe) substitutions and one rvWF mutated in the RGD domain of the C-terminal part of vWF-subunit (Asp1746Gly) were also studied. Among the various samples tested, only rvWF Asp1746Gly had no affinity for GPIIb/IIIa. In contrast, GPIIb/IIIa similarly bound to the other vWF, independently of the proteic environment, the factor VIII level, the degree of multimerization or the mutation of vWF. Our results indicate that subjects with an abnormal vWF binding to GPIIb/IIIa are probably rare and difficult to target for a specific screening. PMID- 10456468 TI - Glycoprotein IIb/IIIa receptor antagonist tirofiban inhibits thrombin generation during cardiopulmonary bypass in baboons. AB - Platelets play a major role in coagulation mechanisms and anti-GPIIb-IIIa antibodies inhibit tissue-factor induced thrombin generation in in vitro studies. Tirofiban, a nonpeptide selective glycoprotein (GP) IIb/IIIa antagonist, preserves platelet number and function during cardiopulmonary bypass (CPB) in baboons. We tested the hypothesis that platelet inhibition by tirofiban inhibits thrombin generation in vivo. Four groups of baboons (n = 7-12) were perfused for 60 min; all groups received heparin (300 units/kg). The controls received only heparin. The low dose (0.1 microg/kg/min) and high dose (0.3 microg/kg/min) infusion groups received tirofiban for 60 min before and 60 min during CPB. The bolus plus low dose infusion group received a 15 microg/kg bolus before starting CPB and a low dose infusion (0.1 mg/kg/min) only during CPB. At end of CPB, compared to control group (2.99+/-0.36 nM), prothrombin fragment F1.2 levels were lower (p<0.05) in low dose infusion group (1.65+/-0.14 nM, mean +/- SE) and high dose infusion group (1.71+/-0.19 nM), but not bolus plus infusion group (2.69+/ 0.49 nM); they remained significantly lower after protamine administration. At end of CPB, thrombin-antithrombin complex levels were lower in high dose infusion group (40.0+/-11.2 ng/ml, p<0.05) compared to control group (76.2+/-7.3 ng/ml). These studies indicate that tirofiban inhibits not only platelet aggregation but also thrombin generation in vivo during CPB, and that this effect is demonstrable even in the presence of intense heparin anticoagulation. They underscore the important inhibitory effect of GPIIb-IIIa antagonists on thrombin generation. PMID- 10456469 TI - A soluble fibrin standard: comparable dose-response with immunologic and functional assays. AB - A soluble fibrin (SF) preparation has been developed as a potential standard by the Scientific and Standardization Committee for use in assays evaluating in vitro preparations and patient plasma samples. The SF standard was prepared by reaction of factor XIII-free fibrinogen with thrombin, followed by neutralization with hirudin and solubilization of the fibrin in acetic acid. As characterized by SDS-PAGE, the polypeptide chain structure shows the anticipated loss of fibrinopeptides and lack of gamma or alpha chain crosslinking. The standard was added to pooled normal plasma at concentrations from 12.5 microg/ml to 340 microg/ml and tested with four commercially available assays based on immunologic reactions using ELISA or latex agglutination or on t-PA cofactor activity for plasminogen to plasmin conversion. Absolute "soluble fibrin" concentrations were calculated using the manufacturers' calibrators and showed distinct dose-response relationships for each assay. Expression of the results following log transformation produced a series of parallel lines, indicating that this SF preparation can serve as a standard, effectively normalizing the disparate proprietary internal calibrators currently used for each assay. PMID- 10456470 TI - Platelet membrane fluidity in non-insulin-dependent diabetes mellitus (NIDDM) subjects, in subjects with vascular atherosclerotic disease (VAD) and in VAD subjects with NIDDM. PMID- 10456471 TI - Recombinant activated factor VII for false aneurysms in patients with normal haemostatic mechanisms. PMID- 10456472 TI - Inhibitor development in a multitransfused patient with severe haemophilia A. PMID- 10456473 TI - Thrombus precursor protein as a measure of prothrombotic activity in a patient with factor VIII inhibitor treated with activated prothrombin complex concentrates. PMID- 10456474 TI - Different sensitivities of various thromboplastins to two blood collection systems for monitoring oral anticoagulant therapy. PMID- 10456475 TI - Intraindividual variability in plasma viscosity in relation to their utility as a cardiovascular risk factor. PMID- 10456476 TI - Non-specific inhibition of ADP-induced platelet antiaggregation by clopidogrel in vitro. PMID- 10456477 TI - Detection of PF4-heparin auto-antibodies by ELISA. PMID- 10456478 TI - Low rate of warfarin-related major bleeding in patients with recurrent venous thromboembolism. PMID- 10456479 TI - No link between the TFPI V264M mutation and venous thromboembolic disease. PMID- 10456480 TI - A polymorphic variant C1367R of the Werner helicase gene and atherosclerotic diseases in the Japanese population. PMID- 10456481 TI - The insertion/deletion polymorphism in the t-PA gene does not significantly affect outcome of meningococcal disease. PMID- 10456482 TI - Quantitative drug interactions prediction system (Q-DIPS): a computer-based prediction and management support system for drug metabolism interactions. AB - OBJECTIVE: Drug biotransformation and interactions are a major source of variability in the response to drugs. The superfamily of cytochromes P450 plays a key role in this phenomenon but, because of the complexity of interactions between drugs and isozymes, it becomes more and more difficult for clinicians to master the knowledge required to predict the occurrence of such drug interactions. To predict and help manage the occurrence of cytochrome P450 dependent interactions, we developed an original computer application: Q-DIPS (quantitative drug interactions prediction system). METHODS: A multidisciplinary work team was created, associating clinical pharmacologists, pharmacists and a computer scientist. Major steps of investigation were: (1) the creation of a database to collect qualitative and quantitative data describing substrates, inhibitors and inducers of specific cytochrome P450 isozymes, with quality assessments; (2) the development of multi-access to these data and (3) their incorporation into extrapolation systems allowing the prediction of in vivo drug interactions on the basis of in vitro data. As an example, prediction and validation studies of CYP3A4 inhibition by ketoconazole and fluconazole will be discussed. RESULTS: Q-DIPS gives up-to-date information, in dynamic tables, describing which specific P450 isozymes metabolise a given drug, as well as which drugs may inhibit or induce a given isozyme. To better answer common clinical questions and help to rapidly evaluate the risk of interactions, it is possible to obtain an overview of substances causing interactions with a specific drug or to focus on drugs taken by a patient ("clinical case"). For each question, key references, relevant quantitative data and quality indices are easily accessible. Two modules allowing input with commercial names and the anatomical therapeutic chemical classification were also included. On the basis of enzymatic and pharmacokinetic data generated in vitro or collected in vivo, the extrapolation module integrates quantitative models to predict the impact of a treatment on enzymatic activities. The simplest model predicted a strong but fluctuating inhibition of CYP3A4 by ketoconazole, whereas the impact of fluconazole was lower. Validations with published in vivo data suggested an appropriate prediction of the risk. CONCLUSION: The current Q-DIPS prototype shows promising potential for helping to improve the management of drug interactions involving metabolism. Validation of extrapolation techniques need to be completed, in view of including important factors such as intrahepatocyte drug accumulation, contribution of metabolites to inhibition as well as in vitro non-specific binding to microsomal proteins. The final goal will be to help select the most judicious clinical studies to be performed so as to avoid useless, expensive and unethical investigations in man. PMID- 10456483 TI - A systematic review of vinpocetine therapy in acute ischaemic stroke. AB - OBJECTIVES: To determine whether vinpocetine decreases short- and long-term case fatality and proportion of dependent survivors if administered within 2 weeks of stroke onset. METHODS: All published and unpublished trials were attempted to be identified using the standard search strategy of the Cochrane Collaboration Stroke Review Group, using MEDLINE searches performed with all known manufacturer code names and trade names of vinpocetine and by contacting manufacturers of vinpocetine to give information of all randomised controlled trials on vinpocetine in stroke. Researchers who participated in trials on vinpocetine in Hungary were asked for further information. Only truly randomised, unconfounded clinical trials that compared the effect of vinpocetine to either placebo or another reference treatment for acute stroke where treatment started no later than 14 days after stroke onset were eligible for inclusion. Data synthesis and analysis was performed using the Cochrane Review Manager software (RevMan version 3.0). RESULTS: Among the identified studies on vinpocetine in stroke, only one fulfilled the selection criteria for inclusion in the review. No death occurred in the study groups and no statistically significant difference was found in dependency between the treatment and the placebo groups. No adverse effects were reported. CONCLUSIONS: Based on only one small randomised controlled unconfounded study, presently there is not enough evidence to decide whether the administration of vinpocetine does or does not decrease case fatality and dependency in acute stroke. PMID- 10456484 TI - Bronchoprotective and bronchodilator effects of single doses of (S)-salbutamol, (R)-salbutamol and racemic salbutamol in patients with bronchial asthma. AB - OBJECTIVES: The drug salbutamol is used as a 50: 50 racemic mixture of its two enantiomers, (R)- and (S)-salbutamol. Previous studies suggest that the (R) enantiomer is active, and the (S)-enantiomer is either inert or may be responsible for adverse effects. The aim of the study was to measure the protection given against methacholine (MCh) and adenosine monophosphate (AMP) by (R)-, (S)- and rac-salbutamol and their bronchodilator effects. METHODS: A double blind, placebo-controlled, four-way cross-over study was performed in subjects with mild to moderate asthma. There were three groups: AMP30 (n = 10), MCh30 (n = 13) and MCh180 (n = 10). The groups received AMP or MCh challenges at either 30 min or 180 min after each of four pretreatments: 100 microg (S)-salbutamol, 100 microg (R)-salbutamol, 200 microg rac-salbutamol or placebo (normal saline), each administered via nebuliser. Spirometry was measured at 30, 60, 90, 120, 150 and 180 min in the MCh180 group. RESULTS: (R)- and rac-salbutamol showed equivalent bronchoprotective effects at 30 min. PC20AMP increased by 3.22 (1.86) and 3.41 (2.15) doubling doses (P < 0.001) and PC20MCh increased by 2.86 (1.09) and 2.75 (0.89) (P < 0.001) respectively. (S)-salbutamol caused no equivalent effect. There was no significant effect at 180 min. No hyper-responsiveness occurred after treatment with (S)-salbutamol. The mean increase in forced expiratory volume in 1 s (FEV1) was 12.4% (6.8%) with (R)- and 12.0%(7.7%) with rac salbutamol at 90 min. No significant change in FEV1 occurred with (S)-salbutamol. CONCLUSIONS: These results confirm other recent findings that the bronchoprotective and bronchodilator effects of salbutamol are attributable to its (R)-enantiomer. No adverse effects were noted after single doses of (S) salbutamol. PMID- 10456485 TI - Multidrug resistance modulation in vivo: the effect of cyclosporin A alone or with dexverapamil on idarubicin pharmacokinetics in acute leukemia. AB - OBJECTIVE: To determine the effect of the coadministration of the multidrug resistance (MDR) modulators cyclosporin A (CyA) alone or plus dexverapamil (D Ver) on idarubicin (IDA) pharmacokinetics in patients with acute leukemia. METHODS: Pharmacokinetic studies were performed in 27 patients with a diagnosis of acute myelogenous leukemia (AML), who were being treated with a combination chemotherapy regimen including idarubicin and cytarabine for the induction of a first remission (n = 14), or of a second remission (n = 7), or for remission consolidation (n = 6). Of these 27 patients, nine were coadministered CyA and seven were coadministered CyA plus D-Ver as MDR modulators. Blood was sampled at appropriate intervals after each of the three IDA daily administrations. IDA and idarubicinol (IDAOL) were assayed by HPLC. Pharmacokinetic evaluations were performed by means of a two-compartment open model with zero-order absorption and first-order elimination using the WinNonlin pharmacokinetic software package. RESULTS: CyA markedly increased the area under the concentration time-curve (AUC) of both IDA [558.26 (197.25) microg x h x l(-1) vs 315.44 (158.28) microg x h x l(-1); P < 0.01] and IDAOL [2896.60 (736.38) microg x h x l(-1) vs 1028.49 (603.95) microg x h x l(-1); P < 0.001] when coadministered as a single modulator, due to a lower total body clearance (CL) [83.51 (52.44) l x h(-1) x m( 2) vs 139.65 (69.45) l x h(-1) x m(-2); NS]. When patients received two MDR modulators simultaneously (D-Ver plus CyA), IDA exposure was essentially the same as in those of the no inhibitor group [331.29 (95.49) microg x h x l(-1) vs 315.44 (158.28) microg x h x l(-1); NS], whereas the IDAOL total body exposure was greater than in the no inhibitor group [2030.32 (401.11) microg x h x l(-1) vs 1028.49 (603.95) microg x h x l(-1); P < 0.01], even if less than in patients receiving CyA as a single MDR modulator (IDA + CyA group) [AUC 2030.32 (401.11) microg x h x l(-1) vs 2896.60 (736.38) microg x h x l(-1); P < 0.05], suggesting an antagonistic effect against those of CyA on IDA and IDAOL elimination and/or an unpredictable redistribution. The main pharmacokinetic parameters of IDA, such as CL and volume of distribution at steady state (Vdss), were remarkably affected by the coadministration of CyA or CyA plus D-Ver, but no statistically significant difference was noted because of IDA pharmacokinetic interpatient variation. CONCLUSION: The results show that CyA alone at a dose of 10 mg x kg( 1) daily significantly increased systemic body exposure to both IDA and IDAOL in acute leukemia, and suggest that these pharmacokinetic effects were at least partially decreased when D-Ver was coadministered with CyA. Our findings raise important questions concerning the need for a dosage adjustment of IDA when MDR modulators are coadministered. PMID- 10456486 TI - Influence of serum protein binding on the uptake and retention of idarubicin by sensitive and multidrug resistant human leukemic cells. AB - OBJECTIVE: The objectives of the investigations were (1) to determine the binding characteristics of idarubicin (IDA) in human serum and cell culture solutions, (2) to determine the effect of protein binding on the uptake and retention of IDA by human leukemic cell lines in culture and the extent to which R-verapamil (R VRP), an inhibitor of the P-glycoprotein (P-gp) transporter, can modulate these processes, and (3) to assess the importance of protein binding on cytostatic and chemosensitizer action in vivo. METHODS: The protein binding of IDA was determined using equilibrium dialysis. Cell uptake of IDA was measured using sensitive and P-gp-containing resistant human leukemic cell lines (HL-60 and HL 60-Vinc) in vitro. IDA was assayed spectrophotofluorometrically. RESULTS: In the incubation media examined, the free fraction of IDA varied more than seven-fold from approximately 60% in 15% fetal calf serum (FCS)/PBS to only 8% in human serum. Cellular uptake of IDA was approximately three times higher in medium containing low protein concentrations. R-VRP eliminated the difference in IDA uptake between resistant and sensitive cell lines and this was the case when the cells were incubated in solutions containing both high and low protein concentrations. However, R-VRP did not overcome the effect of high protein concentrations on IDA uptake. CONCLUSIONS: Plasma protein binding is an important determinant for cellular uptake of IDA in vitro. This should be taken into account when interpreting results of in vitro functional assays with patient material. Chemosensitizers such as R-VRP are effective in both high and low protein solutions. Investigations like these may be useful for evaluating cytostatic efficacy and chemosensitizer action in vivo. PMID- 10456487 TI - Fluvoxamine is a potent inhibitor of tacrine metabolism in vivo. AB - OBJECTIVE: In vitro studies have shown that tacrine is metabolized by cytochrome P4501A2 (CYP1A2). One of the monohydroxy-metabolites has been incriminated with tacrine-induced hepatotoxicity. The aim of this study was to establish whether the potent CYP1A2 inhibitor fluvoxamine in clinically relevant doses could inhibit tacrine metabolism. METHODS: Eighteen healthy young men were enrolled in an open, randomized crossover study. In the first study period a single oral dose of tacrine 40 mg was given. In the second period the volunteers were randomized to maintenance doses of fluvoxamine 50 or 100 mg per day, and a single oral dose of tacrine 20 mg was given. RESULTS: Fluvoxamine was found to be a very potent inhibitor of tacrine metabolism. A fractional decrement in tacrine clearance of approximately 85% was found with both fluvoxamine doses, which was in good agreement with a prediction based on in vitro data. The medians of the steady state concentration of fluvoxamine were 43 nM (range 25-49) and 70 nM (range 44 124) in the 50 mg per day and 100 mg per day groups, respectively. The steady state concentration of fluvoxamine correlated with the fractional decrement in tacrine clearance (Spearman Rs = 0.53, P < 0.05). Modest, but statistically significant, reductions in the formation of the metabolites 1- and 2 hydroxytacrine were found during concomitant fluvoxamine treatment. CONCLUSION: Fluvoxamine at clinically relevant doses is a potent inhibitor of tacrine metabolism. This interaction is very likely to have clinical relevance. Whether concomitant fluvoxamine treatment reduces tacrine-induced hepatotoxicity needs further study. PMID- 10456488 TI - Pharmacokinetic interactions between microemulsion formulated cyclosporine A and diltiazem in renal transplant recipients. AB - OBJECTIVE: Bilateral cyclosporin A (CsA) and diltiazem pharmacokinetic interactions have previously been investigated, however, not with the new microemulsion preconcentrate formulation of CsA (Sandimmun Neoral). In addition, the pharmacokinetic effects on the pharmacological active metabolites of diltiazem have not previously been investigated. We performed a pharmacokinetic interaction study in renal transplant recipients, measuring both unmetabolised CsA and diltiazem in addition to three of the main metabolites of diltiazem (MA, M1, M2). METHODS: Nine CsA-treated renal transplant patients were treated with diltiazem, 90-120 mg b.i.d., for 4 weeks. Pharmacokinetic investigations were performed both before and at the end of the diltiazem treatment period. Six non CsA-treated renal transplant patients served as controls of CsA interactions with diltiazem and its metabolites. RESULTS: Diltiazem treatment resulted in a significant mean increase in the area under the concentration time curve (AUC) for CsA of 51(8)% (P < 0.008) and a peak concentration (Cmax) of 34(8)% (P < 0.05), without altering time to peak concentration (tmax). CsA, however, did not significantly influence diltiazem pharmacokinetics, though two of the metabolites (M1 and M2) tended to be increased. CONCLUSIONS: Diltiazem interacts significantly with the pharmacokinetics of CsA in the new microemulsion formulation. Microemulsion-formulated CsA, however, did not show significant interaction with diltiazem pharmacokinetics. PMID- 10456489 TI - CYP2D6 and GSTM1 genotypes in a Polish population. AB - OBJECTIVE: The aim of the present study was to investigate the distribution of the CYP2D6 and GSTM1 genotypes in a Polish population. METHODS: One hundred and forty-five unrelated healthy individuals from the western region of Poland were studied. The CYP2D6 genotype was analysed by means of polymerase chain reaction (PCR) amplification for the CYP2D6*3 and CYP2D6*4 alleles. The GSTM1 genotype was also analysed by means of a PCR assay to determine two genotypes: GSTM1-1 (positive) and GSTM1-0 (negative). RESULTS: Fourteen subjects (9.6%) were classified as poor metabolisers. The frequency of CYP2D6*4 and CYP2D6*3 was 23.1% and 2.1%, respectively. The frequency of GSTM1 nulled genotype in a Polish population came to 49%. CONCLUSION: The frequencies of poor metabolisers for CYP2D6 and GSTM1 nulled genotype among a Polish population were similar to those observed in other Caucasian populations. PMID- 10456490 TI - Grapefruit juice has no effect on quinine pharmacokinetics. AB - OBJECTIVE: As quinine is mainly metabolised by human liver CYP3A4 and grapefruit juice inhibits CYP3A4, the effect of grapefruit juice on the pharmacokinetics of quinine following a single oral dose of 600 mg quinine sulphate was investigated. METHODS: The study was carried out in ten healthy volunteers using a randomised cross-over design. Subjects were studied on three occasions, with a washout period of 2 weeks. During each period, subjects received a pretreatment of 200 ml orange juice (control), full-strength grapefruit juice or half-strength grapefruit juice twice daily for 5 days. On day 6, the subjects were given a single oral dose of 600 mg quinine sulphate with 200 ml of one of the juices. Plasma and urine samples for measurement of quinine and its major metabolite, 3 hydroxyquinine, were collected over a 48-h period and analysed by means of a high performance liquid chromatography method. RESULTS: The intake of grapefruit juice did not significantly alter the oral pharmacokinetics of quinine. There were no significant differences among the three treatment periods with regard to pharmacokinetic parameters of quinine, including the peak plasma drug concentration (Cmax), the time to reach Cmax (tmax), the terminal elimination half-life (t1/2), the area under the concentration-time curve and the apparent oral clearance. The pharmacokinetics of the 3-hydroxyquinine metabolite were slightly changed when volunteers received grapefruit juice. The mean Cmax of the metabolite (0.25+/-0.09 mg l(-1), mean +/- SD) while subjects received full strength grapefruit juice was significantly less than during the control period (0.31+/-0.06 mg l(-1), P < 0.05) and during the intake of half-strength grapefruit juice (0.31+/-0.07 mg l(-1), P < 0.05). CONCLUSION: These results suggest that there is no significant interaction between the parent compound quinine and grapefruit juice, so it is not necessary to advise patients against ingesting grapefruit juice at the same time that they take quinine. Since quinine is a low clearance drug with a relatively high oral bioavailability, and is primarily metabolised by human liver CYP3A4, the lack of effect of grapefruit juice on quinine pharmacokinetics supports the view that the site of CYP inhibition by grapefruit juice is mainly in the gut. PMID- 10456491 TI - The effect of cimetidine on the steady-state pharmacokinetics and pharmacodynamics of warfarin in humans. AB - OBJECTIVE: The interaction of multiple oral doses of cimetidine on the steady state pharmacokinetics and pharmacodynamics of warfarin was investigated in six healthy male volunteers. METHODS: The subjects were given individually adjusted doses of warfarin to achieve therapeutic levels of prothrombin activity. The established daily maintenance oral dose of warfarin was kept stable throughout the trial and, on study days 8-14, each volunteer received a 800-mg daily dose of cimetidine. The degree of anticoagulant response produced by warfarin was quantified by the determination of both the prothrombin time and factor-VII clotting activity. RESULTS: Cimetidine co-administration had no significant effect on the pharmacokinetics of the more potent S-warfarin but significantly increased by 28% (P < 0.05) mean R-warfarin trough plasma concentrations and decreased by 23% (P < 0.05) mean R-warfarin apparent clearance. Both prothrombin time and factor-VII clotting activity displayed considerable inter-subject variability and were not significantly affected by concurrent cimetidine treatment. The reduction of apparent clearance of R-warfarin by cimetidine was found to be the effect of inhibition of the formation of warfarin metabolites as determined by apparent formation clearance values (+/-SD) of R-6-hydroxywarfarin (31.1+/-7.4 ml/h baseline; 18.5+/-4.5 ml/h at end of cimetidine treatment; P < 0.01), and R-7-hydroxywarfarin (6.9+/-1.3 ml/h baseline; 4.3+/-1.1 ml/h at end of cimetidine treatment; P < 0.01). CONCLUSION: Cimetidine stereoselectively affects the steady-state pharmacokinetics of warfarin by inhibiting the disposition of the less potent R-warfarin in humans. However, this interaction is likely to be of minimal clinical significance in most patients. PMID- 10456492 TI - Grapefruit juice increases the bioavailability of artemether. AB - OBJECTIVE: To evaluate the effect of grapefruit juice on the pharmacokinetics of artemether in plasma and saliva after a single oral dose and to detect concentration-dependent electrocardiographic changes (bradycardia and QTc prolongation). METHODS: Six healthy male subjects were given a standard breakfast followed by two tablets of 50-mg artemether administered with water; 1 week later, the tablets were administered with 350 ml double-strength fresh frozen grapefruit juice. For 8 h, 17 blood- and saliva samples were collected, and 17 electrocardiograms were recorded. Drug and metabolite concentrations were measured by means of high-performance liquid chromatography with electrochemical detection. The pharmacokinetic parameters were determined using a one-compartment model. RESULTS: Grapefruit juice significantly (P = 0.001) increased the mean peak concentration (Cmax) of artemether more then twofold from 42 (SD 17) ng/ml to 107 (28) ng/ml. The time to reach Cmax (tmax) with grapefruit juice was 2.1 (0.6) h compared with 3.6 (17) h with water (P = 0.02). The area under the concentration time curve (AUC) almost doubled with grapefruit juice from 177 ng x h/ml to 336 ng x h/ml (P = 0.003). The elimination half-life remained unchanged (1.0 h vs 1.3 h). No major changes in the kinetics of the metabolite dihydroartemisinin were detected. Low artemether levels and zero dihydroartemisinin levels were found in saliva. No influences of artemether were observed on 17 electrocardiograms during the 8 h after drug intake in particular there were no signs of bradycardia or QTc prolongation. CONCLUSION: Grapefruit juice significantly increases the oral bioavailability of artemether without an effect on the elimination half-life. It suggests a role for intestinal CYP3A4 in the presystemic metabolism of artemether. PMID- 10456493 TI - Polypharmacy among older men in South Wales. AB - OBJECTIVE: To describe the pattern of polypharmacy (PP) among older men and to relate medication use to personal, social and medical information. METHODS: Information on medication use, both prescribed and 'over the counter' (OTC), was collected from 1906 men, aged 56-75 years, observed on up to four occasions since 1979 in a community survey the Caerphilly prospective study. On each visit, a variety of questionnaires regarding personal, social and medical factors were completed, and a brief medical examination was conducted. Medication use was related to some of the questionnaire information and biological measurements collected in order to identify factors associated with PP. RESULTS: A quarter of the men (475/1906) reported using three or more prescription-only medicines (PoMs), with 9% (163) using five or more (major PP). PP was related to increasing age, lower social class, not being in employment, smoking and obesity (high body mass index). Men with a medical history, especially of high blood pressure, angina, heart attack, or hospital admission in the last 5 years, comprised a large proportion of those on major PP. Higher levels of PoM use by this group had been apparent over the previous 14 years. Men on PP reported lower levels of self rated health and higher rates of non-PoM use. Cardiovascular and, to a lesser extent, central nervous and respiratory system drugs were the main medicines used by men on major PP. CONCLUSIONS: PP is common among men aged 56-75 years in Caerphilly, South Wales. It is related to many personal, social and medical factors, and associated with lower self-rated health status and greater use of non-PoMs. Cardiovascular medicines are the main contributor to major PP. Those on PP require regular review and, where possible, PP should be reduced as it has many potential adverse effects. PMID- 10456494 TI - Safeguarding your electronic identity with encryption. PMID- 10456495 TI - Sites in the United States contaminated with radioactivity. AB - Over the century that radioactive materials have been mined, processed, produced, and utilized, many sites across the United States have become contaminated. Such sites include bases and installations of the Department of Defense, weapons production and research facilities of the Department of Energy, properties under the authority of other Federal agencies, privately-owned and governmental facilities that are licensed by the Nuclear Regulatory Commission and its Agreement States, and sites licensed by or the responsibility of states. This review reports on aspects of work by the Environmental Protection Agency, the Department of Defense, the Department of Energy, the Nuclear Regulatory Commission, and others to identify sites contaminated with radioactive materials. It also describes the principal programs that have been instituted to deal with them. PMID- 10456496 TI - The utility of spot collection for urinary uranium determinations in depleted uranium exposed Gulf War veterans. AB - The utility of spot urine collections for uranium bioassay determinations was examined in a small cohort of depleted uranium exposed Gulf War veterans. Some members of the group are excreting elevated concentrations of urinary uranium resulting from the metabolism of retained metal fragments, the residua of several friendly fire incidents. Uranium determinations were performed on both 24-h timed collections and spot urine samples using kinetic phosphorescence analyzer (KPA) methodology. Results ranged from non-detectable to 30.7 mcg g(-1) creatinine in a 24-h collection. A creatinine-standardized spot sample and a 24-h uncorrected sample both correlated highly (R2=0.99) with a creatinine corrected 24-h collection, presumed to be the best estimate of the urinary uranium measure. This relationship was upheld when the population was stratified by uranium concentration into a high uranium group (> or = 0.05 mcg U/g creatinine) but for the lower uranium group (< 0.05 mcg U/g creatinine) more variability and a lower correlation was seen. The uncorrected spot sample, unadjusted for volume, concentration or creatinine had the lowest correlation with the 24-h creatinine adjusted result, especially at lower urinary uranium concentrations. This raises questions regarding the representativeness of such a sample in bioassay programs. PMID- 10456497 TI - Correcting for exposure measurement error in a reanalysis of lung cancer mortality for the Colorado Plateau Uranium Miners cohort. AB - The exposure estimates used to date for the analysis of lung cancer mortality in the Colorado Plateau Uranium Miners cohort were developed from radon progeny measurements taken in mines beginning in 1951. Since uranium miners were often exposed over long periods of time and since mines were not continuously monitored, much extrapolation and/or interpolation of measured dose-rates was needed in order to develop estimates of exposure for each of the miners in the cohort. We have recently re-examined the interpolation scheme used to create the histories in the light of the fit of a statistical model for the radon progeny measurements taken in mines within the Plateau, and we have computed revised exposure estimates for the large majority of miners in the cohort. This report describes the use of these new model-based revised exposure estimates in the analysis of lung cancer mortality, using follow-up data current through 1990. Specific issues addressed here are (1) the strength of the association between exposure and risk of lung cancer mortality; (2) effects of attained age and time since exposure upon risk of lung cancer mortality; and (3) exposure-rate effects upon risk. Results using the revised exposure estimates are compared to those obtained fitting the same models using the original Public Health Service (PHS) exposure estimates. We found evidence that the new exposure histories provide a better fit to the lung cancer mortality data than do the histories based upon the original PHS dose-rate estimates. In general, the new results show a stronger overall relationship (larger slope estimate) between lung cancer mortality and exposure per unit exposure compared to those obtained with the original estimates, while displaying similar age at exposure and time since exposure effects. In the reanalysis the impact of low dose-rate exposure is found to be relatively unchanged before and after exposure error correction, while the estimate of the effect of high dose-rate exposure is considerably increased. Even after applying our measurement error corrections, evidence of inverse dose-rate effects is found, since the estimate of the impact of high dose-rate exposure is still below that of the low dose-rates. The magnitude and statistical significance, however, of the dose-rate effect estimates are diminished when fit using the revised exposure estimates. PMID- 10456498 TI - Comparison of the MCNP calculated and measured radiation field quantities near the RB reactor. AB - The RB experimental reactor has operated at Vinca Institute of Nuclear Sciences since the end of April 1958. In this paper, neutron and gamma-ray spectra and corresponding dose quantities near the reactor, calculated by using the MCNP code, are compared to the measured values during the Third International Intercomparison Experiment on Nuclear Accident Dosimetry carried out at the RB reactor in 1973. Discrepancies in the correlation declared power of the reactor dose rates are found. Good agreements are obtained between measured and calculated neutron and gamma-ray spectra, and corresponding absorbed doses in air, but only after the reactor declared power is multiplied by a correction factor, determined in this study. PMID- 10456499 TI - Probabilistic approach to ranking sources of uncertainty in ELF magnetic field exposure limits. AB - There is a need to improve the biological data, dosimetry, and risk assessment methodology used for setting guidelines for occupational exposures to extreme-low frequency magnetic fields. This paper illustrates how a probabilistic approach can be used to determine priorities for future research based upon the analysis of biological and dosimetric variables that affect stimulation of the heart by magnetically-induced currents. A model was constructed to predict a level of whole-body magnetic-field exposure below which no cardiac stimulation is expected (Bncs). For each iteration of the model, a value was selected from cardiac stimulation threshold, shape factor, and conductivity distributions by Latin Hypercube sampling, and a value for Bncs was computed. The very wide range of simulated Bncs magnetic-field values obtained indicates that there is considerable uncertainty about what constitutes a "safe" level of exposure. The results show that the major occupational-exposure guidelines are very conservative with respect to risks of cardiac stimulation. The minimum Bncs value computed (0.01 T) by either a circular-loop model or an ellipsoid dosimetric model is ten times the highest recommended workday exposure value in a guideline. The lowest 5% Bncs value calculated for a circular-loop model is about 50-times the ICNIRP occupational exposure limit; the lowest 5% Bncs value calculated for an ellipsoid model is more than 100 times the ACGIH occupational exposure limit. A new finding is that the method specified by several guidelines for determining guideline compliance when exposures occur at multiple frequencies (additive weighting of harmonics) leads to substantially lower Bncs estimates relative to a probably more valid rms-weighting method. Probabilistic sensitivity analyses indicate that lack of knowledge of the threshold for cardiac stimulation is the greatest source of uncertainty as to what is a "safe" level of exposure to extreme-low-frequency magnetic fields. Additional research to address this source of uncertainty may be expected to have the greatest potential impact to reduce overall uncertainty about safe magnetic field exposures. PMID- 10456500 TI - Uncertainty of the tritium dose conversion factor. AB - Environmental releases of tritium oxide at a number of Department of Energy nuclear weapons facilities contribute to a significant portion of environmental dose. Several conversion factors are utilized in the estimation of human impact from these releases, e.g., dispersion coefficients, consumption rates, uptake factors, transport factors, dose conversion factors, and risk coefficients. A probabilistic determination of the tritium dose conversion factor was generated in this work to assess the uncertainty of the internal dosimetry required to estimate dose equivalent given an intake of tritium oxide. The tritium dose conversion factor was found to vary by a factor of about 15 with a median value of 2.2 x 10(-11) Sv Bq(-1) when considering orthovoltage x rays as the standard for estimating the relative biological effectiveness of tritium. The median dose conversion factor increases by about 50%; however, when gamma radiation is considered as the standard. The current deterministic estimate of the tritium dose conversion factor published by the DOE and the EPA is 1.7 x 10(-11) Sv Bq( 1), 25-50% lower than the median probabilistic values. The tritium oxide dose conversion factor model was found to be most sensitive to biological half-life and quality factor and is highly dependent on the standardizing radiation for RBE assessments. PMID- 10456501 TI - The effect of moisture on the adsorption of xenon by activated carbon. AB - The following procedure was used to determine the effect of moisture on the adsorption of xenon from air onto activated carbon: A known amount of water is added to a sample of dried activated carbon and allowed to equilibrate. This activated carbon is then split between two cylindrical beds placed in a temperature controlled water bath, and air is passed through the beds sequentially. Because the beds contain pre-moistened activated carbon from the same sample, the first bed acts as a buffer, maintaining a constant humidity in the second. The mean holdup time of a pulse of 133Xe injected into the second bed is used to determine the adsorption coefficient for xenon under these conditions. Measurements were made for three carbons activated to 35, 40, and 59%, respectively, at temperatures of 25 degrees C and 55 degrees C. The effect of moisture on the same activated carbon at these two temperatures shows an affine relationship that could be helpful in extending these results to other temperatures. At low moisture uptakes, a plot of the log(adsorption coefficient) vs. moisture uptake gives a straight line. PMID- 10456502 TI - IAEA/EPA international climatic test program for integrating radon detectors. International Atomic Energy Agency/Environmental Protection Agency. AB - As an element of the joint IAEA-EPA International Radon Metrology Evaluation Program, a climatic test of long-term integrating radon detectors was conducted at the U.S. EPA Radiation and Indoor Environments National Laboratory. The objective of this study was to test the performance of commonly used commercially available long-term 222Rn detector systems under extreme climatological conditions using filtered polycarbonate CR-39 plastic analyzed by the manufacturer using the track-etch method, unfiltered LR-115 film analyzed by the manufacturer, and Teflon based electrets analyzed in the field by EPA using the manufacturer's equipment. The EPA environmental radon chambers were used to expose detectors to extreme cold and dry (less than 4.0 degrees C air temperature and 25% relative humidity) and hot and humid (greater than 35 degrees C air temperature and 85% relative humidity) climatic conditions. During phase I detectors were exposed to low temperatures and low humidities, and during phase II detectors were exposed to high temperatures and high humidities. Typical indoor equilibrium fractions (near 50%) and radon concentrations of about 150 Bq m(-3) were maintained for each phase, which lasted 90 d. The results indicated that the optimal detector for extreme climatic conditions is dependent on the relative importance of bias and precision. Overall, however, the filtered track etch type detector produced the most reliable results under the extreme conditions. PMID- 10456503 TI - Limits and uncertainties--with special regard to radon measurements. AB - The concept of limits was introduced to assure a commonly tolerated low health risk from the exposure to radioactivity or chemicals. In most countries concentration limits are given by the authorities and are fixed to certain values. The question is how to prove compliance with limits, keeping in mind that all measurements are subject to uncertainties. This means there is always a certain probability for a wrong decision. Generally, the authorities demand only the answer "below" or "above" the limit and will not accept statements on probability. Advantages and disadvantages of different decision making philosophies are discussed. PMID- 10456504 TI - Screening level dose assessment of aquatic biota downstream of the Marcoule nuclear complex in southern France. AB - Aquatic biota in the Rhone River downstream of the Marcoule nuclear complex in France are exposed to natural sources of radiation and to radioactivity released from the Marcoule complex. A simple conservative screening level model was used to estimate the range of concentrations in aquatic media (water, sediments, and aquatic organisms) of both artificial and natural radionuclides and the consequent absorbed (whole body) dose rates for aquatic organisms. Five categories of aquatic organisms were studied, namely, submerged aquatic plants (phanerogam), non-bottom-feeding fish, bottom-feeding fish, mollusca, and fish eating birds. The analysis was based on the radionuclide concentrations reported in four consecutive annual radioecological monitoring reports published by French agencies with nuclear regulatory responsibilities. The results of this assessment were used to determine, qualitatively, the magnitude of any potential health impacts on each of the five categories of aquatic organisms studied. The range of dose rate estimates ranged over three orders of magnitude, with maximum dose rates estimated to be in the order of 1 to 10 microGy h(-1). These maximum dose rates are a factor 40 or more below the international guideline intended to ensure the protection of aquatic populations (about 400 microGy h(-1)), and a factor ten or more below the level which may trigger the need for a more detailed evaluation of potential ecological consequences to the exposed populations (about 100 microGy h(-1)). As a result, chronic levels of radioactivity, artificial and natural, measured in aquatic media downstream of Marcoule are unlikely to result in adverse health impacts on the categories and species of aquatic organisms studied. Thus, based on the screening level analysis discussed in this paper, a more detailed evaluation of the dose rates does not appear to be warranted. PMID- 10456505 TI - Assessment of need for transport tubes when continuously monitoring for radioactive aerosols. AB - Aerosol transport tubes are often used to draw aerosol from desirable sampling locations to nearby air sampling equipment that cannot be placed at that location. In many plutonium laboratories at Los Alamos National Laboratory, aerosol transport tubes are used to transport aerosol from the front of room ventilation exhaust registers to continuous air monitors (CAMs) that are mounted on nearby walls. Transport tubes are used because past guidance suggests that extraction of aerosol samples from exhaust locations provides the most sensitive and reliable detection under conditions where the rooms have unpredictable release locations and significant spatial variability in aerosol concentrations after releases, and where CAMs cannot be located in front of exhaust registers without blocking worker walkways. Despite designs to minimize particle loss in tubes, aerosol transport model predictions suggest losses occur lowering the sensitivity of CAMs to accidentally released plutonium aerosol. The goal of this study was to test the hypotheses that the reliability, speed, and sensitivity of aerosol detection would be equal whether the sample was extracted from the front of the exhaust register or from the wall location of CAMs. Polydisperse oil aerosols were released from multiple locations in two plutonium laboratories to simulate plutonium aerosol releases. Networked laser particle counters (LPCs) were positioned to simultaneously measure time-resolved aerosol concentrations at each exhaust register (representative of sampling with transport tubes) and at each wall-mounted CAM location (representative of sampling without transport tubes). Results showed no significant differences in detection reliability, speed, or sensitivity for LPCs positioned at exhaust locations when compared to LPCs positioned at the CAM wall location. Therefore, elimination of transport tubes would likely improve CAM performance. PMID- 10456506 TI - Cohen's paradox. PMID- 10456507 TI - Response to Cohen's comments on the Lubin rejoinder. PMID- 10456508 TI - IAEA safeguards implementation. International Atomic Energy Agency. PMID- 10456509 TI - A long-term, controlled-outcome analysis of in utero versus neonatal cleft lip repair using an ovine model. AB - Successful open repair of a cleft lip in utero has the advantage of scarless wound healing in the fetus. Unfortunately, no long-term outcome studies have been performed to evaluate the efficacy of these repairs. Moreover, no study to date has compared the long-term results of an in utero cleft lip repair to a similar, control-matched, newborn cleft repair. This study was performed to evaluate the 9 month outcome of in utero cleft lip surgery compared with an identical cleft lip repair performed on infant lambs. In utero epithelialized cleft lips were created through an open hysterotomy in sixteen 65-day-old fetal lambs (term = 140 days) using methods described by Longaker et al. Eight of 16 animals underwent subsequent in utero repair of these clefts at 90 days gestational age. The repair of the remaining eight animals was delayed until 1 week postpartum. At 9 months, the animals were analyzed for changes in lip contour and for the degree of scarring by hematoxylin and eosin and Masson's trichrome collagen staining. Two animals in each group died from preterm labor. Of the animals that survived to term, all repaired lips had some degree of abnormality postoperatively. One of six lips repaired in utero dehisced before delivery. Three of six neonatal repairs dehisced in the first postoperative month. In the remaining animals with intact lip repairs, the vertical lip height on the repaired side was an average of 9 to 12 mm shorter than the normal lip in both the in utero and neonatally repaired animals. Phenotypically, the postnatally repaired animals had more lip distortion and visible notching. Histologically, the in utero repair was scarless and the neonatal repairs had scar throughout the entire vertical height of the lip with an associated loss of hair in this region. Maxillary growth was also evaluated. There was no inhibition of maxillary growth in the animals that underwent in utero cleft lip repair. However, in the neonatal repair group, significant maxillary retrusion was evident. Compared with the cleft side of the maxilla, horizontal growth was decreased by 11 percent (p = 0.01). Compared with the intrauterine repair group, there was a 17-percent decrease in horizontal maxillary width (p = 0.01). Straight-line in utero repair of a cleft lip produces a better long-term result in terms of maxillary growth than a similar repair performed postnatally in the ovine model. There was no diminution in maxillary growth in the animals treated in utero. Histologically, in utero repair of clefts was indeed scarless. However, both lip repairs produced lips that were significantly shorter than their contralateral noncleft sides. This degree of lip shortening would require a secondary lip revision, thereby defeating the purpose of performing an intrauterine repair. Comparisons now need to be made between in utero and neonatal repairs using a Millard-type rotation advancement technique before intrauterine treatment can be considered to be more beneficial than our current treatment modalities. PMID- 10456510 TI - Nonsurgical correction of nasal deformity in unilateral complete cleft lip: a 6 year follow-up. AB - Nasal deformity in unilateral cleft lip and palate patients increases with time, tongue malposition being one of the causes. Some authors have emphasized the role of nasal and adjacent facial musculature as active extrinsic agents. Another cause of alar deformity can be the lack of a proper foundation because of a maxillary hypoplasia in the region of the pyriform foramen. If alar collapse occurs, the septum bends convexly toward the cleft side. Tissues are soft and plastic during the neonatal period. Once the infant is about 3 months of age, it becomes difficult to correct the nasal deformity. Therefore, any resource used from the first day, and mainly during the first 15 days of life, will be useful to prevent the increasing deformity and to avoid the surgical correction. A controlled clinical trial was planned to compare the anthropometric measurements of the nasal region in two series of patients with unilateral complete cleft lip. In the first group, we included 44 patients who came to our clinic during the first 2 days of life and the second group consisted of 47 patients who were more than 15 days of age at the time of the first consultation. To provide control data for the evaluation of the results after 6 years of follow-up in both series of cleft patients, we also included a third group of 48 healthy 6-year-old children. A nasal component added to the occlusal prostheses was only used in the first group up to the time of surgery. The same surgeon performed a Millard II procedure with muscular reposition as described by Delaire in all the patients. Nasal measurements taken with a caliper, obtained directly from plaster models by using surface impressions of the babies, were confirmed by a laser three dimensional measuring device. The statistical comparison between both series showed a significant increase of the columellar length in the first group. A 6 year follow-up to compare growth and cosmetic results of the nose revealed a better and permanent nasal nostril symmetry and no alar cartilage luxation in the patients who had had the nasal component. These results highlight the importance of the early treatment and allow us to suggest the nasal prostheses as a way to prevent the increasing nasal deformity, to help nasal remodeling, to obtain columellar elongation, and to avoid or decrease the need for primary surgery of the cleft nose. PMID- 10456511 TI - Use of the nasal SMAS to reconstruct defects of the tip, alae, columella, or septum. AB - Reconstruction using traditional methods for small and medium-sized losses of the soft tissues of the lower third of the nose, including the tip, the columella, alae, or the cartilaginous septum, has meant leaving cicatricial stigmata surrounding the region and has required at least two operational stages. A comprehensive study of the local anatomy was completed, and a specific technique (presented herein) was used to transfer and distribute the well-vascularized layer of superficial muscle and aponeurotic tissue (SMAS) from the dorsum of the nose to the needed site, which provided bulk and a well-nourished surface that could be covered by a distant skin graft, using a one-step flap. The surgical approach to the SMAS was carried out through the scar over the site of tissue loss or through the site of regional ablation or the traumatic wound. Several representative cases are presented. PMID- 10456512 TI - Full-thickness grafting of acute eyelid burns should not be considered taboo. AB - Split-thickness skin grafts are commonly used for the treatment of acute eyelid burns; in fact, this is dogma for the upper lid. Ectropion, corneal exposure, and repeated grafting are common sequelae, almost the rule. It was hypothesized that for acute eyelid burns, the use of full-thickness skin grafts, which contract less than split-thickness skin grafts, would result in a lower incidence of ectropion with less corneal exposure and fewer recurrences. The records of all patients (n = 18) who underwent primary skin grafting of acutely burned eyelids (n = 50) between 1985 and 1995 were analyzed retrospectively. There were 10 patients who received full-thickness skin grafts (12 upper lids, 8 lower lids) and 8 patients who received split-thickness skin grafts (15 upper lids, 15 lower lids). Three of 10 patients (30 percent) who received full-thickness skin grafts and 7 of 8 patients (88 percent) who received split-thickness skin grafts developed ectropion and required reconstruction of the lids (p = 0.02). No articles were found substantiating the concept that only split-thickness grafts be used for acute eyelid burns. The treatment of acute eyelid burns with full thickness rather than split-thickness skin grafts results in less ectropion and fewer reconstructive procedures. It should no longer be considered taboo and should be carried out whenever possible and appropriate. PMID- 10456513 TI - Orbital floor and infraorbital rim reconstruction after total maxillectomy using a vascularized calvarial bone flap. AB - A number of techniques have been introduced to support the orbital floor after maxillectomy without orbital exenteration. These methods include skin graft or muscular sling, but they have resulted in severe complications, such as enophthalmos, global ptosis, diplopia, and facial deformity. Currently, advanced microvascular reconstruction using bone and soft tissue is performed by many surgeons. This usually results in the filling of the postmaxillectomy defect, but the lack of support for the orbital rim and floor by the bone flap may still cause the complications mentioned above. Vascularized calvarial bone flap was chosen in this study for reconstruction of the orbital floor and infraorbital rim to function as a buttress, to reconstruct recipient sites of poor vascular bed after radiation therapy, and to withstand further postoperative radiation. By providing a solid floor and rim, these complications can be prevented with satisfactory function and aesthetically acceptable results. From September of 1995 to July of 1998, we performed vascularized bone flap for the reconstruction of the orbital floor and infraorbital rim in four cases after total maxillectomy involving the orbital floor. With a follow-up period from 19 to 35 months (mean, 27 months), we obtained significant improvement of functional and aesthetically acceptable results without global ptosis, enophthalmos, diplopia, or severe facial contour deformity. PMID- 10456514 TI - Functional hemitongue reconstruction with the microvascular ulnar forearm flap. AB - Thirteen patients with squamous cell carcinoma of the tongue underwent full thickness longitudinal resection of the hemitongue and immediate microvascular reconstruction using a large, contoured ulnar forearm flap. Six of the 13 patients had a composite resection for which an additional vascularized iliac crest graft was used to reconstruct the mandible and to provide support to the overlying contoured flap. To increase tongue mobility, the skin flap was designed for independent reconstruction of the hemitongue and the floor of mouth. Twelve patients were evaluated for swallowing and speech, including dietary assessment, cineradiography, and voice spectrographic analysis. Contrast cineradiography was performed to determine oral tongue mobility during the first phase of swallow. Nine patients with a narrow reconstructed tongue root and a large surface area in the floor of the mouth had good tongue mobility, allowing them to transfer food dynamically from the mouth into the pharynx for swallowing. The remaining three patients, who had a wide tongue root and an ill-defined floor of the mouth, had decreased tongue mobility and poor oral transport. The functional outcome of swallowing and speech strongly correlated with the shape of the root of the tongue, the proximity of the reconstructed tongue to the palate, and the surface area of the floor of the mouth. PMID- 10456515 TI - The deposition of talc in patients with silicone gel-filled breast implants. AB - This study was initiated to understand why talc was found in the entrance wounds and pericapsular scars of patients with silicone breast implants. Twenty-five surgical gloves made between 1977 and 1992 by a major glove manufacturer were evaluated with infrared spectroscopy. Seven gloves manufactured before early 1983 contained talc. Later manufactured gloves contained calcium carbonate instead of talc. Talc-containing gloves were submitted to independent laboratories for scanning electron microscopic examination and x-ray microanalysis to obtain finer detail. Remnants of the mold-release agent talc were found in the matrix of the patient-contact side of the glove. Wetting studies showed that silicone oil (gel bleed) immediately wet the surface of rubber glove. After donning, silicone oil was massaged between the thumb and index finger of each washed, talc-containing glove for 1 minute and then daubed from the glove to a microscopic slide. The slide was viewed with polarized light microscopy for the presence of talc crystals. Transfer of talc occurred using silicone oil, but not saline, in each talc-containing glove. Three factors were likely involved in the greater talc deposition that occurred with patients undergoing silicone gel-filled breast implantation than in patients undergoing other surgical procedures. (1) The wetting of the gloves with silicone gel bleed from implants loosened the talc in the glove matrix. (2) Hand-intensive surgical use freed the talc from the gloves. (3) Loose talc adhered to the silicone elastomer of the breast implants and was deposited with implant placement. PMID- 10456516 TI - Postmastectomy breast reconstruction in Connecticut: trends and predictors. AB - Using the population-based Connecticut Tumor Registry, postmastectomy breast reconstruction was examined in the 10,756 breast cancers in Connecticut women (including 10,133 in white women and 554 in black women) diagnosed from 1988 to 1995. Reconstruction increased from 6.4 percent of cancers in 1988 to 9.1 percent in 1991, but it declined to 4.7 percent of cancers in 1992 (when the Food and Drug Administration instituted a restriction on the use of silicone gel implants); by 1995, the rate had recovered to 8.5 percent. Reconstruction was negatively associated with age, poverty rate of the census tract of residence, and black (versus white) race; these associations require further study. PMID- 10456517 TI - A comparison of pedicled and free TRAM flaps for breast reconstruction in a single institution. AB - Several reports concluded that free tissue transfer of the transverse rectus abdominis muscle (TRAM) flap for breast reconstruction is superior to pedicled transfer of the flap. In an effort to compare the various parameters of both techniques, the authors took advantage of a unique experience at one hospital where one surgeon (D.L.L.) used only the pedicled method and the other (N.J.Y.) used only free tissue transfer. Additionally, the authors compared the findings of the study with the experiences of other surgeons by surveying active members of the American Association of Plastic Surgeons. The records of 119 patients who underwent TRAM flap reconstruction between January of 1988 and July of 1997 were reviewed. Of these, 33 patients received free TRAM flaps, and 86 received pedicled TRAM flaps. To provide an adequate number of patients for statistical analysis, only those with unilateral, single-muscle reconstructions were considered (immediate or delayed). This provided 61 patients in the pedicled flap group and 26 in the free tissue group. Parameters examined included length of operation and of hospitalization, amount of pain medication used, amount of blood lost and received, and complications. A small subset of the patients had hospital records available to compare hospital charges; the comparison of 17 pedicled and 12 free TRAM flaps showed a mean difference of $15,637 (p < 0.001) in favor of the pedicled flap. On the basis of the findings from this study, it seems that the pedicled TRAM flap has significant economic and clinical advantages over the free TRAM flap. There is less need for blood, a shorter operating time and hospital stay, and a need for less pain medication. However, both methods of transfer have indications and contraindications in certain clinical settings. It will always remain the responsibility of the surgeon to evaluate all issues and select a method that is economically responsible and within the abilities of the surgeon, while producing a satisfactory outcome that best serves the patient. The information provided in this report should aid in accomplishing this goal. PMID- 10456518 TI - Radiotransparency of the triglyceride mammary prosthesis: a quantitative analysis with mastectomy specimens. AB - Breast cancer is the most frequent malignant tumor in women. It is estimated that 10 percent of women will present with a breast cancer during their lives. It is well known that mammography is the best technique for the early diagnosis of nonpalpable tumors, thus improving life expectancy. However, mammary prostheses may hide between 23 and 82 percent of the normal mammary tissue in mammography, and thus may delay the diagnosis of malignant mammary tumors, making prognosis worse. To solve this problem, oil-filled prostheses have been developed. In this study, 14 mastectomy specimens were used. Mammograms of the tissue pieces alone and also mammograms of the tissue pieces covering a 270-cc Trilucent prosthesis were used to verify whether the prosthesis allows observation of malignant signs in mammography. Mammograms were evaluated by an independent experienced radiologist. The following variables were studied: number of mammograms necessary to examine each specimen; kilovoltage and milliamperage necessary for each mammogram; number of microcalcification groups (malignant); number of macroscopic calcifications (benign); and rarefaction areas that were suspected for malignancy. All of these variables were measured for both mammograms for which the mastectomy specimens were covering and those for which the specimens were not covering the prothesis. Finally, the kilovoltage and milliamperage increases necessary to visualize the mammograms with mastectomy specimens covering the prosthesis were determined. Statistical analysis of the results obtained was performed. There were no significant differences in the number of mammograms (p = 0.391), the number of microcalcifications (p = 0.890), the number of macrocalcifications (p = 0.239), and finally in the presence of rarefaction areas (p = 1.000) observed in the mammograms in specimens either covering or not covering the prosthesis. However, there were significant differences (p < 0.001) between the kilovoltage and milliamperage applied to carry out the mammograms of specimens with and without the prosthesis. Thus, Trilucent prostheses allow visualization of the microscopic and macroscopic calcifications as well as rarefaction areas in mammograms. However, these mammograms required a higher kilovoltage and milliamperage compared with specimens not covering the prosthesis. To explore the whole gland, it might be necessary to perform two series of mammograms: one to detect the area shadowed by the prosthesis and one to observe the rest of the peripheral gland. PMID- 10456519 TI - Integrated breast mound reduction and nipple reconstruction with the wraparound flap. AB - A new method for nipple reconstruction is described that combines revision of an autologous tissue breast mound with creation of a projecting nipple. The method is applicable only to reconstructed breast mounds that must be reduced or lifted to achieve symmetry with the opposite breast. In this technique, the mound is reduced as if it were a normal breast, using an inverted-T or vertical mammaplasty pattern. In this way, breast projection can be increased and, if necessary, the inframammary fold can be elevated. A rectangular flap is created from skin and subcutaneous tissue that would normally be discarded during the breast reduction, and this flap is wrapped around on itself to form a projecting nipple. This new technique avoids the flattening of the breast mound usually seen after nipple reconstruction because it does not take tissue away from the completed breast mound to make the nipple. In appropriate patients who require reduction in size of their reconstructed breast mound, the wraparound flap nipple reconstruction is worth considering. PMID- 10456520 TI - Keloids have continuous high metabolic activity. AB - The adenosine triphosphate (ATP) content of keloids and scars resected from patients was demonstrated by high-performance liquid chromatography. The surface color of hypertrophic scars was red or pink and that of atrophic scars was white. The ATP content of red scars was (in mmol/g protein) 1.06 +/- 0.14, of pink scars 0.12 +/- 0.02, of white atrophic scars 0.19 +/- 0.06, and of keloids 1.06 +/- 0.19. The longer the elapsed time after the trauma, the lower the level of ATP in scar tissues (correlation coefficient = -0.506; p = 0.005 by Spearman's rank correlation). However, ATP levels in keloids were still high 10 years after the injury. Fibroblasts and fibrocytes in keloids and scars were counted in histologic preparations stained with hematoxylin and eosin. The average number of fibroblasts in a definite area (56 x 10(-4) mm at a magnification of x400) was 4.8 in keloids, 5.1 in red scars, 2.4 in pink scars, and 1.3 in white atrophic scars. The number of fibrocytes in the same area was 0.4 in keloids, 0.4 in red scars, 2.3 in pink scars, and 1.3 in white atrophic scars. These results indicate that keloids and red hypertrophic scars have higher ATP levels and contain more fibroblasts than pink or white scars, and they also suggest that the levels of ATP and the number of fibroblasts decrease when red hypertrophic scars change into atrophic scars. In keloids, ATP and fibroblasts seem to remain at high levels for a long time. PMID- 10456521 TI - Instability of the distal radio-ulnar joint: stabilization by a Gore-Tex ligament. AB - Diagnosis and treatment of problems caused by instability of the distal radio ulnar joint are complex. In this retrospective study, the results of a uniform procedure by using a Gore-Tex ligament for stabilization of the distal radio ulnar joint are presented. In eight patients, nine wrists were operated on. The mean age of the patients was 35 years. The dominant side was involved in six patients. The mean follow-up was 3.4 years. Three of nine wrist problems were of spontaneous onset. In these wrists, the final results were excellent. The other six wrists were operated on for problems after trauma: four patients after soft tissue injury, one patient after distal radius fracture, and one patient after forearm fracture. Among these cases, one result was excellent, four were good, and another one was fair. PMID- 10456523 TI - The direct effect of intraorbital pressure on orbital growth in the anophthalmic piglet. AB - Tissue expanders placed within the orbit can have a positive effect on orbital and ipsilateral midfacial growth. To date, there is no precise method for controlling and monitoring expansion to induce normal growth in the developing facial skeleton. The present study was undertaken to determine the optimal physiologic pressure required to stimulate normal orbital growth and to determine whether above-normal growth could be achieved with higher intraorbital pressures. Using a neonatal swine model, an accurate method of monitoring intraorbital pressure, precisely controlling intraorbital expansion, and achieving normal orbital growth was explored. Sixteen male, 3-week-old Yorkshire piglets were randomly divided into three surgical groups. In each group, the left orbit was the experimental side, and the contralateral right orbit served as an untreated control. Group 1 (n = 6) underwent enucleation only. Group 2 (n = 5) underwent enucleation and orbital expansion at a near-normal physiologic pressure of 20 mmHg. Group 3 (n = 5) underwent enucleation and orbital expansion at a supernormal pressure of 60 mmHg. Spherical tissue expanders (10 cc) with a separate injection port were utilized as the orbital expanders. Pressure was monitored by an electronic manometer that was calibrated daily. Morphology of the orbits was documented by photography, the dimensions of the orbits were quantitated by three-dimensional mechanical digitization, and orbital volumes were calculated. In the unexpanded, anophthalmic control group, a significant reduction in radial growth after evisceration was seen. In group 2, the orbit stimulated with a consistent pressure of 20 mmHg, just above the physiologic normal pressure of 17 mmHg, showed an increase in radial dimension of 8 percent compared with the unoperated side. In the high-pressure group of 60 mmHg, an increase of 16 percent in the radius was observed over the 4-week period. This led to a corresponding increase in orbital volumes with increased pressure. Utilizing a paired t test, these differences in the radial and volumetric growth of the orbit were statistically significant (p < 0.005). The results obtained demonstrated a direct relationship between intraorbital pressure and the growth of the bony orbit in the radial dimension. On the basis of this study, we concluded that orbital expansion maintained at normal physiologic pressure can stimulate normal orbital growth in the neonatal facial skeleton. In addition, application of above-normal pressures for expansion can induce accelerated orbital growth. PMID- 10456522 TI - Osseointegration of preformed polymethylmethacrylate craniofacial prostheses coated with bone marrow-impregnated poly (DL-lactic-co-glycolic acid) foam. AB - Osseointegration of bone marrow-PLGA-coated, preformed polymethylmethacrylate cranioplasties offers the possibility of reducing: operative time, periimplant seroma and infection, metallic fixation, and periprosthetic resorption following surgical skull remodeling. These alloplastic materials are FDA-approved but previously have not been used together to promote cranioplasty incorporation. The objective of this study was to determine whether the use of PLGA foam coating improves host osseointegration of preformed, textured, polymethylmethacrylate prosthetic cranioplasties. A critical-sized cranial defect was created in two groups of 10 and one group of three rabbits. The defect was filled with either a textured, preformed polymethylmethacrylate disc or a textured, preformed polymethylmethacrylate disc coated with poly (DL-lactic-co-glycolic acid). Both implants were immersed in autologous bone marrow for 20 minutes before implantation. Half of each group of 10 were killed at 3 weeks, and the remainder at 6 weeks. A third group of three rabbits with excised periosteum was evaluated at 6 weeks. Histologic analysis of the discs determined relative amounts of cancellous bone formation adjacent to the prostheses. Woven trabecular bone was present at each host bone to implant perimeter interface at 3 weeks, with fine fibrous capsular formation around the implants. Thicker, lamellar trabeculae were present at 6 weeks with an increased fibrous layer surrounding both types of implants. Bone formed on the superficial and deep implant surfaces in a noncontiguous fashion. Two of five measures showed that total bone formation was significantly greater in the PLGA-coated implants. Polymethylmethacrylate discs coated with bone marrow-impregnated PLGA foam demonstrate increased bone formation at 3 and 6 weeks as compared with non-coated preformed polymethylmethacrylate discs. Only implants with preserved periosteum showed bone formation away from the host-implant interface (centrally) on the superficial surface at 6 weeks. PMID- 10456524 TI - Treatment of posttraumatic ocular dysmotility using autogenous buccal fat grafts in a porcine model. AB - Diplopia occurring after orbital trauma is a complex and difficult clinical problem. Numerous potential mechanisms exist by which it may occur. Restrictive ocular dysmotility caused by intraorbital scarring is a major component in diplopia's pathogenesis. The current large animal study was conducted to develop an experimental model of restrictive ocular dysmotility that would quantitatively characterize the biomechanical properties of the globe rotations. Using this model, a novel method of restoring the low-friction milieu within the orbit by interposing a buccal fat graft was tested. In the initial stage, the baseline force duction was measured in 20 pig eyes using a highly sensitive, digital tensiometer. Traumatic violation of Tenon's fascia with electrocautery into the extraconal fat and the periorbita was followed by direct suturing of the extraocular muscle to the nearest orbital periosteum. After 6 weeks, the measurements (again in the field of the traumatized muscle) were repeated, and the eyes were divided into two treatment groups (n = 10 eyes per group). The left eye received the standard lysis of adhesion, whereas the right eye received lysis and buccal fat interposition grafting. The third and final force measurements were performed 6 weeks after treatment. The results showed a baseline linear load displacement curve of 0 to 8 mm, with the globe rotating 400 microm for every 1000 mg of tensile load. Surgical trauma increased the slope as defined by load/displacement but, surprisingly, the relationship remained linear in the entire range from 2 to 8 mm. This linear relationship was seen in all stages: baseline, after trauma to Tenon's fascia, after surgical lysis alone, and after lysis with buccal fat interposition. The difference was in the slope, or stiffness. Lysis alone partially reduced the slope, but it was still higher than baseline. Lysis and buccal fat grafting returned the slope to near baseline. This, however, did not reach the level of statistical significance. It seems that a focal intervention along the course of an extraocular muscle altered the composite behavior of orbital resistance to globe rotation. Although buccal fat grafting did not significantly improve motility, it did not worsen it. PMID- 10456525 TI - Effects of recombinant human growth hormone on the development of burn scarring. AB - Anabolic agents, such as recombinant human growth hormone (rhGH), have been used effectively to ameliorate the catabolic response to burn injury and to improve wound-healing. However, in experimental animal models, growth hormone has also been associated with increased renal scarring. The effect of rhGH on the development of human scarring is unknown. Therefore, the purpose of this study was to assess the effect of rhGH on the scarring of human skin after burn injury. A series of 94 patients was studied in a prospective randomized double-blind clinical trial. Patients receiving 0.2 mg/kg/day subcutaneous rhGH during their acute hospital stays presented with the same quality and intensity of scarring as patients receiving a placebo. Similar reconstructive needs also resulted. The treatment of severely burned children with recombinant human growth hormone during the acute-phase hospital course did not increase scarring of the burn wound. PMID- 10456526 TI - Muscle flaps' triphasic microcirculatory response to sympathectomy and denervation. AB - Whether sympathectomy and somatic denervation in muscle flaps increased microcirculatory flow in the short or long term, thus producing an effect similar to the delay phenomenon, which increases survival in transferred skin flaps, was determined. The rat cremaster muscle flap model was used for in vivo microscopy. In the left cremasters of 30 Sprague-Dawley rats, the genitofemoral nerve was divided and the proximal vessels were stripped of their adventitia. The muscle was not elevated. In each rat, the contralateral cremaster served as the control. The rats were assigned to one of five groups: no delay before observation, a 24 hour delay, a 48-hour delay, a 7-day delay, or a 14-day delay. After the delay, red blood cell velocity, vessel diameters, number of functional capillaries, and leukocyte-endothelial interactions were measured. Microvessel response to topical vasoactive substances was measured. Immediately after denervation, red blood cell velocity increased transiently (71 percent; p = 0.006). Main arterioles dilated (20 percent; p = 0.02) at 24 hours, and capillary perfusion increased 36 percent (p = 0.001) at 2 weeks. The microvessels had hyperactive responses to all vasoactive agents 2 weeks after denervation. These findings indicate that proximal sympathectomy with somatic denervation leads to a triphasic, dynamic response in the peripheral microcirculation of the cremaster muscle flap. An initial acute hyperadrenergic phase was followed by a nonadrenergic phase, with significant vasodilatation, and a sensitized phase, with increased capillary perfusion and hyperresponsiveness to vasoactive substances. This study shows that with minimal access to the cremaster muscle flap neurovascular pedicle and without changing the blood supply to the flap, significant hemodynamic improvements can be made in the peripheral microcirculation. PMID- 10456527 TI - Hypoxia regulates VEGF expression and cellular proliferation by osteoblasts in vitro. AB - Numerous studies have demonstrated the critical role of angiogenesis for successful osteogenesis during endochondral ossification and fracture repair. Vascular endothelial growth factor (VEGF), a potent endothelial cell-specific cytokine, has been shown to be mitogenic and chemotactic for endothelial cells in vitro and angiogenic in many in vivo models. Based on previous work that (1) VEGF is up-regulated during membranous fracture healing, (2) the fracture site contains a hypoxic gradient, (3) VEGF is up-regulated in a variety of cells in response to hypoxia, and (4) VEGF is expressed by isolated osteoblasts in vitro stimulated by other fracture cytokines, the hypothesis that hypoxia may regulate the expression of VEGF by osteoblasts was formulated. This hypothesis was tested in a series of in vitro studies in which VEGF mRNA and protein expression was assessed after exposure of osteoblast-like cells to hypoxic stimuli. In addition, the effects of a hypoxic microenvironment on osteoblast proliferation and differentiation in vitro was analyzed. These results demonstrate that hypoxia does, indeed, regulate expression of VEGF in osteoblast-like cells in a dose dependent fashion. In addition, it is demonstrated that hypoxia results in decreased cellular proliferation, decreased expression of proliferating cell nuclear antigen, and increased alkaline phosphatase (a marker of osteoblast differentiation). Taken together, these data suggest that osteoblasts, through the expression of VEGF, may be in part responsible for angiogenesis and the resultant increased blood flow to fractured bone segments. In addition, these data provide evidence that osteoblasts have oxygen-sensing mechanisms and that decreased oxygen tension can regulate gene expression, cellular proliferation, and cellular differentiation. PMID- 10456528 TI - A simplified vertical reduction mammaplasty: shortening the learning curve. AB - Although the vertical reduction mammaplasty is effective in reducing scarring, the technique has not achieved widespread acceptance in North America. There are several reasons for this. Some believe that the vertical reduction mammaplasty is only applicable to smaller breast reductions and that the learning curve is difficult. This article describes modifications to the standard Lejour vertical reduction mammaplasty that simplify the technique and make it more reliable and easier to perform. These modifications include using a medial (or lateral) dermoglandular pedicle, not undermining the skin, using liposuction only rarely to reduce breast volume, and not using pectoralis fascia sutures; the modified technique has been used in a series of 400 vertical breast reductions. In this series, scarring was reduced and the technique was easily learned and applied. It is useful for both small and large breast reductions, with a series average of 525 g removed per breast (range, 100 to 1425 g). By using these modifications, scarring in reduction mammaplasty was effectively reduced while the nipple and areola were safely transposed on a medial or lateral dermoglandular pedicle. Incorporating these technical modifications in a vertical reduction mammaplasty is recommended as a method of improving results and limiting complications. PMID- 10456529 TI - Vertical mammaplasty: early complications after 250 personal consecutive cases. AB - No surgeon likes to face complications. It takes effort to treat them personally and more effort to note, count, analyze, and demonstrate them. The author carefully followed 250 personal consecutive patients (476 breasts) who underwent vertical mammaplasties between 1990 and 1998; studying the complications and their relationship with the types of breasts and patients was very instructive. The main observations from this study follow. The most frequent benign complication was seroma (5 percent of breasts), which usually required one or two aspirations after surgery. Hematomas occurred in six patients (1.2 percent of breasts), who had all had mastopexies. Hematomas required immediate surgical evacuation. The major complication of breast reduction, i.e., areola necrosis, was rare (only two partial necroses occurred), but it left deformities that were difficult to correct. Infection without tissue necrosis was rare (two cases), and healing complications happened in only 5.4 percent of all cases. Healing complications were directly related to the size and fat content of the breasts. None occurred in mastopexy cases. For reductions, delayed skin healing was observed in 5 percent of cases and delayed breast tissue healing in 3 percent of cases. More healing complications occurred after liposuction of the breast, which was performed in the more fatty breasts. Delayed healing of skin and breast tissue was bothersome because healing was slow, but it left only a moderate deformity. In cases of delayed healing, frequent dressings, rinsing the wound with antiseptic solutions, giving antibiotics if needed, and refraining from early surgical intervention are the keys to success. Good personal contact with the patient, especially if healing is slow, is the best way of helping her and avoiding aggressive attitudes. In conclusion, this survey revealed few complications; however, it does show that the risk of delayed and slow healing is greater in larger breasts. In obese patients, a simpler operation may be indicated, such as liposuction with skin reduction alone or a free nipple graft, as long as the patient is not motivated to obtain the best possible result. PMID- 10456530 TI - Vertical mammaplasty: update and appraisal of late results. AB - Vertical mammaplasty was evaluated after 10 years of experience and the study of 250 personal consecutive cases, and a few minor technical modifications are presented. In the beginning, the main advantage of the technique seemed to be the reduced amount of scarring, as this technique avoids submammary scars. With more experience, the major advantages seem to be its adaptability to breasts of various sizes and shapes and its good, consistent, and stable results. Liposuction is useful and reliable, but it cannot be applied to all fatty breasts because fat is sometimes intimately mixed with parenchyma and cannot be reached by a blunt cannula. Vertical mammaplasty raises many questions among surgeons. The most frequent were collected and answered in this report. PMID- 10456531 TI - Primary epithelioid sarcoma of the scalp. AB - This article retrospectively reviewed a case of epithelioid sarcoma of the scalp; a treatment plan for this type of neoplasm has not been well defined in the literature because of the rarity of sarcomas in general and sarcomas located in the head and neck in particular. No comparative results can be drawn when dealing with such lesion in the scalp. Early recognition with an aggressive approach to confirm the existence of an epithelioid sarcoma is mandatory; a high index of diagnostic awareness is needed to recognize this uncommon tumor. Early diagnosis can only be auspicious. Early wide surgical resection is imperative to ensure better control of imperceptible tumor extension, and well-planned diagnostic and therapeutic intervention, rather than isolated management and referral for adjunctive treatment, is important. PMID- 10456532 TI - Macrocystic schwannoma in the seventh cervical nerve. AB - We have recently treated a case of asymptomatic macrocystic schwannoma discovered between the left neck and the supraclavicular fossa. The tumor, originating from the left seventh cervical nerve, was extirpated. Nearly 70 percent of the tumor area showed macrocyst with transparent fluid collection. The tumor was diagnosed as macrocystic schwannoma histopathologically. Schwannoma is often associated with small cysts but rarely with macrocysts. Because schwannoma is a benign tumor, partial resection against an attempt at total removal resulting in progressive neurologic deficits is recommended. In general, schwannoma does not present any apparent symptoms at an early stage, because it occurs and progresses very slowly. Schwannoma originating at the spinal nerve root has been called a dumbbell-shaped tumor because of its characteristic shape on MRI and cystogram images. We recently have treated a case of dumbbell-shaped tumor that was an asymptomatic macrocystic schwannoma in the seventh cervical nerve. PMID- 10456533 TI - Total mandibular and lower lip reconstruction with a prefabricated osteocutaneous free flap. AB - Large, complex bony defects can be a vexing problem for the reconstructive surgeon, especially when standard donor sites are not available or do not provide sufficient tissue. Using the concept of flap prefabrication, we demonstrated in a single patient that (1) iliac crest bone chips and bone morphogenic protein in an alloplastic mandibular tray can ossify in a heterotopic location and (2) neovascularization sufficient to support a large, custom-designed bone graft occurs within a convenient "carrier" flap. Ultimately, the fields of angiogenesis and osteogenesis research could significantly contribute to the ability of the plastic surgeon to construct the "ideal" composite prefabricated flap for complicated reconstruction. PMID- 10456534 TI - Painful pectoralis major myospasm as a result of sternal wound reconstruction: complete resolution with bilateral pectoral neurectomies. AB - We report a patient with a highly unusual and previously unreported complication with the use of the pectoralis major muscle to treat the infected median sternotomy. The diagnosis of painful myospasm was made by a combination of physical findings and exclusion of other conditions such as recurrent infection. Treatment by pectoral denervation was relatively simple and highly successful. Patients with chest-wall pain after sternal wound reconstruction should have myospasm entertained as a possible cause. PMID- 10456535 TI - Minimally invasive harvest of the gracilis muscle. AB - Acceptance of minimally invasive plastic surgery has been predicated on decreasing morbidity while maintaining the quality and costs of outcomes. The major patient complaint about the gracilis muscle donor site has almost solely been related to the length of the thigh scar, and thus would appear to be an ideal indication for outcome improvement using minimally invasive techniques. A method of endoscopically assisted gracilis muscle harvest, therefore, was developed, starting with a transverse incision just proximal to the knee to identify the gracilis tendon. This endoscopic port allows retrograde subfascial dissection of the muscle and precise identification of its anatomic course, whereupon a small proximal medial thigh incision can be made secondarily for direct access to the vascular pedicle. This variation has now been used successfully in 10 patients. The mean proximal thigh scar length was 8.30 +/- 0.74 (SD) cm, and total surgical scars measured 11.84 +/- 0.95 cm, compared with 27.73 +/- 9.55 cm for 16 patients for whom an open method had been used. This diminished scar length was a statistically significant improvement (p < 0.05), verifying the value of the surgical endoscope as an adjunct for harvest of the gracilis muscle as a free flap. PMID- 10456536 TI - Current trends in breast reduction. AB - Current trends in the development of breast reduction surgery include a few minor refinements in classic inverted-T scar methods but, more, the increased use of vertical-scar reduction mammaplasty. The benefits of the latter, which include reduced scar burden and improved long-term projection, are attractive, although the technique itself has proved to be somewhat intuitive and more difficult to master. These shortcomings can be minimized and the technique safely learned by initially applying it to patients with minor degrees of macromastia and ptosis. New modifications and alternative approaches have been introduced recently to address the problematic areas of the vertical-scar technique. A survey of members of the American Society of Plastic and Reconstructive Surgeons was undertaken at the 1998 annual meeting to review the current role of various techniques in breast reduction. The results revealed a slow acceptance of vertical-scar methods and the dominance of the inferior/central pedicle inverted-T scar method for a wide variety of macromastia types. Other issues were reviewed; they revealed the minimal role of both blood transfusion and liposuction and that more than half of breast reductions are still performed on an inpatient basis. PMID- 10456537 TI - Is liposuction safe? PMID- 10456538 TI - Advances in liposuction contouring of calves and ankles. AB - Achieving a predictable, aesthetic result in liposuction contouring of the lower leg is now possible. The evolution of the technique has brought about preoperative, intraoperative, and postoperative advances. This article describes in detail these advances and their rationale. PMID- 10456539 TI - Otoplasty: what I do now--a 30-year perspective. AB - The author presents his otoplasty technique, a variation on Mustarde's original procedure, which he has used for more than 30 years in more than 200 otoplasties. Timing, indications, and photographic documentation are discussed, and the operative procedure, including the preoperative work-up and anesthesia, is described in step-by-step detail. Postoperative care and early and late complications, from infection to the dissatisfied patient, are discussed extensively. PMID- 10456540 TI - Subperiosteal rejuvenation of the midface and periorbital area: a simplified approach. AB - A simplified approach to subperiosteal midface rejuvenation is presented. The technique was developed through experience with 60 subperiosteal face lifts over an 8-year period. Because it does not involve a lower eyelid incision, there is no risk of ectropion, yet the midfacial structures and orbicularis oculi muscle can be elevated easily and precisely. The procedure can be performed alone but is more commonly performed in conjunction with brow lift and traditional SMAS dissection of the lower face and neck. The benefits of this subperiosteal approach to the midface are powerful elevation of the midfacial structures, improvement of the nasolabial folds, enhancement of the cheek prominences, smoothing of the nasojugal folds, and tightening of the orbicularis oculi. The procedure is performed through small incisions in the temple and gingival sulcus. The operation is technically straightforward and poses little risk to neurovascular structures. Concerns over prolonged swelling have not materialized. There have been no instances of lower lid or canthal abnormalities. PMID- 10456541 TI - Office-based skin care and superficial peels: the scientific rationale. PMID- 10456542 TI - Conservation therapy for breast cancer following augmentation mammaplasty. PMID- 10456544 TI - Blindness after reduction of facial fractures. PMID- 10456545 TI - Regional anesthesia on the lacrimal nerve. PMID- 10456546 TI - Xanthelasma procedures. PMID- 10456547 TI - Pure orbital blowout fracture: new concepts and importance of the medial orbital blowout fracture. PMID- 10456548 TI - Long-term follow-up of cranial bone graft and dorsal nasal augmentation. PMID- 10456549 TI - Securing the nostril retainer. PMID- 10456550 TI - Cloth drape fixation using transparent adhesive dressings for facial plastic surgery. PMID- 10456551 TI - Plastic surgery of the face in the fourth century. PMID- 10456552 TI - Direct monitoring using an endoscope for buried flaps. PMID- 10456553 TI - The radial artery perforator free flap. PMID- 10456554 TI - Safe carpal tunnel release with a palmar incision. PMID- 10456555 TI - Postoperative drainage in reduction mammaplasty. PMID- 10456556 TI - Repair of midline incisional hernias. PMID- 10456557 TI - An apology. PMID- 10456558 TI - Steatapheresis: a new term for a new obesity surgery. PMID- 10456559 TI - Peripheral opioid receptors may mediate a portion of the aversive and depressant effect of EtOH: CPP and locomotor activity. AB - Previous investigators have reported that peripheral opioid receptors (located in the gut) may produce aversive effects when activated. Opioid receptors can be activated by endogenous opioids or by-products of ethanol (EtOH) metabolism [e.g., tetrahydroisoquinolines (TIQs)]; both are stimulated following EtOH consumption. Therefore, we tested the hypothesis that a portion of the aversive or depressant effects of EtOH may be mediated through, or modulated by, peripheral opioid receptors. Conditioned place preference (CPP) and locomotor activity were the dependent variables. Prior to EtOH gavage, we antagonized the peripheral opioid receptors with methylnaltrexone (MNTX), an opioid antagonist that does not easily pass through the blood-brain barrier. The effects of EtOH were found to be dose dependent: 1.5 g/kg was hedonically neutral but depressed locomotor activity; 2.25 g/kg EtOH was aversive and also depressed locomotor activity. MNTX (32 mg/kg) treatment was rewarding and stimulated motor activity (especially during the first conditioning session). When combined, 1.5 g/kg EtOH tended to enhance the rewarding effects of MNTX whereas MNTX blocked the aversive effects of 2.25 g/kg EtOH. During the first conditioning session EtOH attenuated the motor stimulant effects of MNTX whereas MNTX antagonized the motor depressant effects of EtOH; there was little effect of MNTX on EtOH-induced motor depression during subsequent conditioning sessions. Pretreatment with various doses of MNTX (0.1, 1.0, 10.0, 32.0 mg/kg) of rats receiving 1.5 g/kg EtOH indicated the effects of MNTX were dose dependent. Drug-induced locomotor activity and time spent in the conditioned compartment were positively correlated, suggesting that both behaviors were homologous. The data suggest that peripheral opioid receptors participate in mediating or modulating a portion of the behavioral effects of EtOH. PMID- 10456560 TI - Beneficial effects of S-adenosyl-L-methionine on aminolevulinic acid dehydratase, glutathione, and lipid peroxidation during acute lead-ethanol administration in mice. AB - Beneficial effects of S-adenosyl-L-methionine (SAM) in preventing inhibition of blood delta-aminolevulinic acid dehydratase (ALAD), alterations in blood and hepatic glutathione (GSH), hepatic and brain malondialdehyde (MDA) formation, and uptake of lead following acute lead plus ethanol coexposure were investigated in mice. Whereas exposure to both lead or ethanol individually produced a significant inhibition of blood delta-aminolevulinic acid dehydratase (ALAD) activity, ethanol administration alone produced only a marginal depletion of hepatic glutathione (GSH). A significant elevation of hepatic MDA concentration was observed following lead or ethanol ingestion. An appreciable increase in brain GSH following ethanol administration whereas a moderate elevation in MDA level following lead plus ethanol administration was observed. Combined lead plus ethanol exposure produced a more pronounced depletion of blood ALAD activity and an increase in hepatic MDA level compared to lead- or ethanol-alone administration. Brain GSH concentration showed an increase compared to untreated control animals or lead-alone-exposed mice. Concomitant administration of SAM partially reversed the inhibition of blood ALAD activity in all three exposed groups (i.e., lead, ethanol, or lead plus ethanol). Lead concentration in blood, liver, and brain was significantly reduced by SAM in lead-alone or lead plus ethanol coexposed groups. The results suggest that supplementation of SAM may have beneficial effects in preventing alterations in some biochemical variables and accumulation of lead in blood, liver, and brain during acute lead plus ethanol exposure in animals. PMID- 10456561 TI - Alcohol, slow wave sleep, and the somatotropic axis. AB - When alcohol is a large proportion of daily nutrient energy, the network of signals for energy homeostasis appears to adapt with abnormal patterns of sleep and growth hormone (GH) release along with gradual acquisition of an addictive physical dependency on alcohol. Early relapse during treatment of alcoholism is associated with a lower GH response to challenge, perhaps reflecting an altered balance of somatostatin (SS) to somatropin releasing hormone (GHRH) that also affects slow wave sleep (SWS) in dependent patients. Normal patterns of sleep have progressively shorter SWS episodes and longer rapid eye movement (REM) episodes during the overall sleep period, but the early sleep cycles of alcoholics have truncated or non-existent SWS episodes, and the longer REM episodes occur in early cycles. During SWS delta wave activity, the hypothalamus releases GHRH, which causes the pituitary to release GH. Alcohol-dependent patients have lower levels of SWS power and GH release than normal subjects, and efforts to understand the molecular basis for this maladaptation and its relation to continued alcohol dependence merit encouragement. More needs to be learned about the possibility of decreasing alcohol dependency by increasing SWS or enhancing GHRH action. PMID- 10456562 TI - Nicotine blocks ethanol and diazepam impairment of air righting and ethanol impairment of maze performance. AB - Results of our previous research in rats demonstrate the following: (a) Angiotensin II (Ang II) inhibits long term potentiation (LTP) in dentate granule cell-perforant path synapses and that this inhibition can be blocked by losartan, an Ang II AT1 receptor antagonist; (b) both ethanol and diazepam inhibit LTP induction and this inhibition can be blocked by losartan; (c) impairment of air righting by ethanol and diazepam (DZ) and eight-arm radial maze performance by ethanol can be blocked by pretreatment with losartan: (d) inhibition of dentate granule cell LTP by Ang II can also be prevented by pretreatment with nicotine. Therefore, it seemed reasonable to hypothesize that ethanol and diazepam impairment of air righting and maze performance might also be blocked by pretreatment with nicotine. The purpose of the present study was to determine the effects of nicotine 0.1, 0.2, 0.3, and 0.4 mg/kg subcutaneously (SC) on 2.0 g/kg ethanol per os (PO) and 1.0 and 2.0 mg/kg DZ intraperitoneally (i.p.) induced impairment of air righting; and to determine if the impaired maze performance due to 2.0 g/kg ethanol PO could be prevented by pretreatment with 0.4 mg/kg of nicotine, SC. Results confirm the hypothesis that moderate doses of ethanol, 2.0 g/kg PO, and DZ, 1.0 mg/kg i.p. impair air righting and that the impairment can be prevented by pretreatment with nicotine SC. Nicotine was not effective in blocking the 2.0 mg/kg DZ impairment of air righting. Nicotine, 0.4 mg/kg SC, prevented the impaired maze performance induced by 2.0 g/kg ethanol PO. PMID- 10456563 TI - Effects of N-methyl-D-aspartate receptor antagonists on reinforced and nonreinforced responding for ethanol in rats. AB - Results of several recent studies indicate that the discriminative stimulus effects of ethanol are related, at least partially, to ethanol-induced decrease in the N-Methyl-D-aspartate (NMDA) receptor function. The role of NMDA receptors in ethanol reinforcement remains still unclear. The aim of the present study was to evaluate the effects of two novel NMDA receptor antagonists in rats lever pressing for 8% ethanol in the oral self-administration procedure. In addition, the effects of the drugs on intensity of nonreinforced responding for ethanol (i.e., "experimental craving") were examined in the extinction procedure. To assess selectivity of the drugs' actions the same range of doses was tested in rats lever pressing for water (control experiments). A low-affinity, uncompetitive NMDA receptor antagonist, MRZ 2/579 (2.5-7.5 mg/kg) selectively and dose-dependently decreased ethanol self-administration. This compound exerted also selective effects on nonreinforced responding for ethanol with lower dose (2.5 mg/kg) increasing and higher dose (5 mg/kg) suppressing operant behavior in the extinction procedure. MRZ 2/579 (5 mg/kg) did not alter open field activity when given in combination with either saline or ethanol (0.5-1 g/kg). In contrast, a glycineB site antagonist, MRZ 2/576 (2.5-7.5 mg/kg) did not produce any selective effects on either reinforced or nonreinforced lever pressing for ethanol. The present results suggest that MRZ 2/579 may selectively suppress both ethanol self-administration and experimental ethanol craving. PMID- 10456564 TI - Effects of haloperidol or SCH-23390 on ethanol-induced conditioned taste aversion. AB - Dopaminergic systems are thought to play an important role in the motivational effects of ethanol. The present experiments examined the effects of haloperidol (a D2 antagonist) and SCH-23390 (a D1 antagonist) on the acquisition of ethanol induced conditioned taste aversion. In four separate experiments, adult male Swiss-Webster mice were acclimated to a 2-h/day water restriction regimen. Subsequently they received four conditioning trials consisting of 1-h access to either 0.2 M NaCl (experiments 1-3) or 0.15 % w/v saccharin (experiment 4). After flavor access on trials 1-3, subjects received either haloperidol (0.1, 0.15, or 0.3 mg/kg), SCH-23390 (0.05 mg/kg), or saline followed 30 min later by 0, 2, or 4 g/kg ethanol. Ethanol-flavor pairings reduced subsequent flavor intakes, indicating the development of conditioned taste aversion. Neither haloperidol of SCH-23390 reduced flavor intakes in the absence of ethanol. However, both haloperidol and SCH-23390 reduced ethanol-conditioned aversion depending on ethanol dose and conditioned flavor. These results are consistent with the notion that dopaminergic processes are important for the development of ethanol-induced conditioned taste aversion, and the notion that dopaminergic receptor systems influence both positive and negative motivational effects of ethanol. PMID- 10456565 TI - Effect of chronic ethanol ingestion on biochemical circadian rhythms in Wistar rats. AB - Chronic ingestion of ethanol for 60 days was known to alter the characteristics of biochemical circadian rhythms in Wistar rats. Peak times of glucose, potassium and lactic acid rhythms were delayed by 18 h, 3 h, and 3 h respectively, whereas peak times of cholesterol and malondialdehyde rhythms were advanced by 3 h and 9 h respectively during ethanol treatment. Significant changes in range (p < 0.001 expect in calcium) and 24 h mean (p < 0.001) of all the biochemical circadian rhythms studied were observed during ethanol treatment. The alterations in the characteristics of these biochemical circadian rhythms could be principally due to the alterations on the hepatic cellular architecture; other plausible underlying reasons are also discussed. PMID- 10456566 TI - The effect of fat and carbohydrate content of the diet on voluntary ethanol intake in golden hamsters. AB - Golden hamsters, which avidly consume ethanol solutions, had continuous access to food and water and to either 15% or 30% ethanol solution (v/v) over a period of weeks. Hamsters consumed approximately equal amounts of absolute ethanol when maintained on either a Purina Chow diet or a high-carbohydrate, low-fat diet (7.6 and 7.1 g/kg/day, respectively), but they consumed substantially less ethanol when maintained on a high-fat, low-carbohydrate diet (4. 4 g/kg/day). However, the pattern of differences among groups of hamsters maintained on different diets was not the same at the two ethanol concentrations. Thus, at the 15% concentration, HC hamsters consumed more than twice as much ethanol as HF hamsters, but at the 30% concentration, the difference in ethanol consumption was greatly reduced and did not prove to be statistically significant. These results indicate that it is important to consider the concentration of the ethanol solution when studying the effects of dietary macronutrient content on ethanol consumption. PMID- 10456567 TI - Taste reactivity in alcohol-preferring AA and alcohol-avoiding ANA rats. AB - Alcohol-preferring rats (Alko alcohol or AA) were tested for taste reactivity to water, sucrose, quinine, and a range of alcohol concentrations (5-40%) both before and after a period of continuous alcohol access. The alcohol-avoiding line of rats (Alko nonalcohol or ANA) was also tested for comparison. It was found that AA rats displayed greater ingestive reactivity to alcohol compared to ANA rats both before and after a three-week period of continuous access to 10% alcohol (during which time AA rats drank significantly more alcohol than ANA rats). AA rats also made significantly more ingestive responses to a 0.3 M sucrose solution and a 0.0005 M quinine solution. Differences between AA rats and ANA rats in aversive reactivity appeared only after the alcohol consumption tests; AA rats made significantly fewer aversive responses to the 30% and 40% concentrations after continuous alcohol access. AA rats also made significantly more aversive responses to the quinine solution. The results suggest that line differences between AA rats and ANA rats in the reactivity response to alcohol solutions have been selected in association with the original selection phenotype of alcohol consumption. PMID- 10456568 TI - Effect of ethanol on (R)- and (S)-salsolinol, salsoline, and THP in the nucleus accumbens of AA and ANA rats. AB - Tetrahydroisoquinolines (TIQ) are active metabolites of dopamine. Intracerebral application stimulates the voluntary ethanol intake. In the present study, the levels of several TIQ's [(S)- and (R)-salsolinol, salsoline and tetrahydropapaveroline (THP)] were measured in the extracellular space of the nucleus accumbens of alcohol-preferring AA and alcohol-avoiding ANA rats. Ethanol (2 g/kg i.p.) caused an increase in dopamine levels in ANA but not in AA rats. Neither (R)- nor (S)-salsolinol concentrations changed after ethanol application, though (S)-salsolinol concentrations were higher in ANA than in AA rats. Ethanol caused an increase in salsoline concentrations in ANA but not in AA rats. THP increased following ethanol, which tended to be stronger in ANA rats. The study revealed differences in the TIQ levels of the nucleus accumbens between AA and ANA rats. In case of changes following ethanol application (dopamine, salsoline, THP), the AA rats were less sensitive. The findings resemble observations in high risk sons of alcoholics with reduced sensitivity to ethanol in young age and increased risk to become alcoholic. PMID- 10456570 TI - Comparison of the proopiomelanocortin and proenkephalin opioid peptide systems in brain regions of the alcohol-preferring C57BL/6 and alcohol-avoiding DBA/2 mice. AB - Differences in the activity of distinct components of the endogenous opioid system between ethanol-preferring and ethanol-avoiding animals may be important in controlling their voluntary alcohol consumption. The objective of the present studies was to compare the activity of two opioid peptide systems (enkephalin and beta-endorphin) in distinct regions of the brain, between the C57BL/6 and DBA/2 mice, using sensitive radioimmunoassays, in situ hybridization, and immunohistochemical techniques. The immunohistochemical studies indicated that there was no significant difference in the number of either beta-endorphin or enkephalin immunopositive cells between the C57BL/6 and DBA/2 mice. The in situ hybridization studies demonstrated a 27% higher content of proopiomelanocortin mRNA in the arcuate nucleus of the C57BL/6 than DBA/2 mice, p < 0.02. The content of proenkephalin mRNA was 25% higher in the nucleus accumbens, p < 0.005 and 23% higher in the caudate putamen, p < 0.01, of the C57BL/6 than DBA/2 mice. There was no significant difference in the content of beta-endorphin peptides in the distinct brain regions investigated. The content of met-enkephalin-arg6-phe7 in the nucleus accumbens and caudate was similar between the two strains of mice, while it was significantly lower in the amygdala, hippocampus, ventral tegmental area, and periaqueductal grey of the C57BL/6 than DBA/2 mice. Thus, there are significant differences in the activity of these two endogenous opioid peptide systems in distinct regions of the brain, between ethanol naive C57BL/6 and DBA/2 mice, which may play a role in controlling their alcohol consumption. PMID- 10456569 TI - Effects of ethanol on the stress-induced expression of NGFI-A mRNA in the rat brain. AB - We investigated expression of NGFI-A/zif268 mRNA, reliable marker for neuronal activation in response to stress in the brain of rats pretreated with ethanol. The rats were orally administrated with either 25% sucrose or 20% ethanol (20 ml of kg body weight) 10 min before the onset of the stress. The rats were exposed to immobilization stress for 20 min and quickly decapitated. The brains were extracted and immediately frozen. The level of NGFI-A mRNA was evaluated by in situ hybridization histochemistry. Prior ethanol administration attenuated the immobilization stress-induced upregulation of NGFI-A mRNA level in the neocortex and hippocampus. However, in the paraventricular hypothalamic nucleus, prior administration of ethanol did not affect the upregulation of this gene. These data suggest that ethanol abolishes the forebrain component of the stress response while it fails to attenuate the stress response on a region of the brain that regulates the autonomic nervous system and the hypothalamo-pituitary-adrenal axis. Regional differences in the expression of NGFI-A mRNA may be important for the complex interactions between ethanol and stress. PMID- 10456571 TI - Infant rats respond differently to alcohol after nursing from an alcohol intoxicated dam. AB - Our previous studies indicate that rat pups are able to detect the low levels of ethanol (175 mg %) found in the milk of a moderately intoxicated dam. The present study tested the effect of infantile interactions (including suckling) with ethanol-treated mothers on later behavioral responsiveness to ethanol's sensory properties. In Experiment 1, pups suckled from dams subjected to a 2.5 g/kg ethanol dose (i.g.) or water-treated females during postnatal days (PDs) 3, 5, 7, 9, 11, and 13. During PD 15, these pups were exposed to procedures to induce a conditioned aversion to the low level of ethanol (175 mg % in water), with lithium chloride as the unconditioned stimulus. Conditioning was more effective for pups with the prior ethanol experience within the nursing context. Greater responsiveness to ethanol in milk also was found for conditioning control pups that had interacted with intoxicated dams than for those that had interacted with water-treated dams. Experiment 2 determined that interaction with an intoxicated dam was sufficient for altered responsiveness to ethanol, in that the additional conditioning procedures of Experiment 1 were not needed for the effect. Generally, a relatively brief history of infantile interaction with ethanol intoxicated dams increased later responsiveness to ethanol's orosensory properties. The results suggest that moderately intoxicated dams within the nursing context provide information to the progeny that may lead to the establishment of ethanol-related memories. PMID- 10456572 TI - Dose and time dependent effects of ethanol on antioxidant system in rat testes. AB - This study was designed to investigate the dose as well as time dependent effects of ethanol on testicular antioxidant defense system in rats. Male Fischer 344 rats were administered ethanol at a dose of 2, 4, and 6 gm/kg orally and control received equal volume of saline and sacrificed 1 h after ethanol ingestion. For time course study, animals were administered ethanol 4 g/kg orally and sacrificed at 1.5, 2, 4, and 6 h after ethanol ingestion. Testicular ethanol concentration increased with increasing doses of ethanol. Copper zinc-superoxide dismutase (CuZn-SOD) activity significantly decreased in the testes of rats treated with increasing doses of ethanol whereas manganese-superoxide dismutase (Mn-SOD) activity significantly increased in a dose dependent manner (181, 186, and 195% of control, respectively). Testicular glutathione (GSH) and malondialdehyde (MDA) levels did not significantly alter with increasing doses of ethanol one hour after ethanol ingestion. Ethanol concentration decreased in the testes with an increase in time after ethanol ingestion. Testicular CuZn-SOD activity significantly decreased whereas Mn-SOD activity increased with an increase in time after ethanol ingestion. Testicular catalase (CAT) activity significantly decreased at 2 h postethanol ingestion. Testicular MDA levels significantly increased at 4 and 6 h after ethanol ingestion indicating that end product of lipid peroxidation. MDA, takes considerable time to form in the testes. A significant decrease in the ratios of CAT/Mn-SOD and glutathione peroxidase (GSH Px)/Mn-SOD in the testes of rat suggests the ability of mitochondria to scavenge reactive oxygen species (ROS). It is suggested that antioxidant enzyme ratios may be used as an important parameter to determine ethanol induced oxidative stress in the tissues. PMID- 10456573 TI - Opposite effects of ethanol and nitrendipine on nicotine-induced emesis and convulsions. AB - The effects of ICV injections were investigated in unanesthetized cats of ethanol alone and in combination with the dihydropyridine calcium antagonist, nitrendipine, on emesis and the convulsions produced by nicotine, which was similarly injected by the ICV route. In the first series of experiments, short lasting convulsions and emesis were the most prominent symptoms after the ICV injection of nicotine in a dose of 1.0 mg. In the second series of experiments the pretreatment of cats with ethanol given ICV in doses of 0.03, 0.2, and 0.3 ml reduced the emesis and prevented the convulsions induced by 1.0 mg dose of ICV nicotine. In the third series of experiments, the ICV injection of nitrendipine in doses of 0.024, 0.16, and 0.24 mg incorporated in the solution of ethanol, given in volumes of 0.03, 0.2, and 0.3 mt, respectively, blocked emesis but not the convulsions induced by the 1.0 mg dose of nicotine given ICV. The results suggest, therefore, that at least two different mechanisms underlie these phenomena. First, the synergistic effects at the neuronal nicotinic ionophores in the brain would act to underlie the antagonistic action of ethanol and nitrendipine on the emetic response. Second, conformational changes brought about by ethanol at voltage-dependent calcium channels in the brain may antagonize the inhibitory effect of the dihydropyridine calcium antagonist, producing the reversal of convulsions. PMID- 10456574 TI - Methanol elevates cytosolic calcium ions in cultured canine cerebral vascular smooth muscle cells: possible relation to CNS toxicity. AB - Acute exposure of cultured canine cerebral vascular smooth muscle cells to methanol (10-400 mM) results in concentration-dependent elevation of the concentration of intracellular free calcium ion ([Ca2+]i) as measured with the fluorescent indicator, fura-2, and digital imaging microscopy. The resting level of [Ca2+]i in the cerebral vascular smooth muscle cells was 89.3+/-5.3 nM. Exposure of these cells to 10 mM methanol for only 5 min resulted in significant elevation in [Ca2+]i (i.e., to 105.7+/-4.6) (p < 0.05). Methanol (10 mM) is a concentration found in the blood of victims demonstrating early CNS toxicity. Other, higher concentrations of methanol rapidly raised [Ca2+]i upwards of 60% over basal resting levels. These result suggest that methanol-induced cerebral vasospasm is a consequence of large rises in intracellular Ca2+. These events could play a crucial role in methanol-induced cerebral edema, brain hemorrhage, and cerebral and retinal infarcts, eventuating in severe deficits in brain blood flow and the known, subsequent CNS disturbances. PMID- 10456575 TI - Mmu and D2 receptor antisense oligonucleotides injected in nucleus accumbens suppress high alcohol intake in genetic drinking HEP rats. AB - Numerous pharmacological and other studies have implicated both Mmu and dopamine receptor subtypes in alcohol consumption. In the genetic drinking rat as well as those chemically induced to drink, evidence has accrued that the abnormal intake of alcohol is underpined by these receptors in the brain. The purpose of this investigation was to demonstrate unequivocally that a biological impairment by antisense oligodeoxynucleotide (ODN) targeted specifically to these two receptor subtypes would disrupt ongoing alcohol drinking. In this project, a new strain of female and male high-ethanol preferring (HEP) rats was used that had free access to preferred concentrations of alcohol over water in a two choice paradigm. A guide cannula for a microinjection needle was first implanted bilaterally above the nucleus accumbens (NAC) of each rat. Following recovery, a dose of either 250 or 500 ng of the Mmu ODN or 500 ng D2ODN was microinjected into the NAC of the rat in a volume of 0.8-1.0 microl. A standard temporal sequence was used in which microinjections were given four times at successive 12-h intervals over a 2-day interval. The control mismatch ODNs corresponding to both the Mmu or D2 receptor antisense were microinjected identically at homologous sites in the NAC. Following the experiments, the brain of each rat was removed and sectioned in the coronal plane for histological analysis so that each microinjection site was identified. The results showed that the Mmu receptor antisense caused a significant dose dependent fall in free access alcohol drinking within 12 to 24 h following the initial microinjection. This decline often persisted for 1 to 2 days in terms of both g/kg intake and proportion of alcohol to water consumed. Similarly, the D2 receptor ODN likewise induced an intense and significant decline in both g/kg and proportion measures of alcohol intake. Since the corresponding mismatch ODN for both Mmu and D2 receptors exerted no effect on either of these measures of alcohol consumption, the specificity of molecular action of the respective antisense molecules on drinking behavior of the HEP rats was confirmed. Thus, these results provide the first unequivocal evidence that the genes for D2 and Mmu receptors are fundamentally involved in abnormal alcohol drinking in the genetically predisposed individual. Finally, important new anatomical evidence is introduced for the critical role of the NAC in the genetic basis of aberrant drinking of alcohol. PMID- 10456576 TI - Commentary on the recommendations of the Royal College of Obstetricians and Gynaecologists concerning alcohol consumption in pregnancy. PMID- 10456577 TI - Reflections on alcohol and the young. AB - Young people's exposure to alcohol in Britain is a major cause of concern to alcohol workers, yet little impact has been made on the problem despite extensive research and numerous recommendations for tackling it. To some extent, this is due to permissive public attitudes and the general lack of knowledge among the public and professionals of the dangers of alcohol misuse, but lack of leadership by government and targetting of alcohol at the young by the drinks industry must take some of the blame. Particular issues of relevance to young people are excessive drinking in pregnancy, which may affect the fetus, alcoholic parents, drinking among 11-15 year olds, and heavy consumption by older adolescents. The pressure that these put on social and health services (and the expense) is considerable. Rather than concentrating resources on treating the damage, greater emphasis should be placed on prevention and detection, for example recognizing risk factors -- immaturity, poor educational performance, antisocial behaviour, mental heath problems and dysfunctional families -- which lead to harm. This will require better education about alcohol for child specialists, teachers, social workers, the police, the legal professions, and others who have to deal with young people. Self-help groups among the young could help spread the message of sensible drinking among their peers. In addition, the number of alcohol agencies in the community which support parents and young people, at present woefully inadequate, need to be increased as a matter of urgency. PMID- 10456578 TI - The synergistic action of ethanol and nerve growth factor in the induction of neuronal nitric oxide synthase. AB - Ethanol alone had no effect on neuronal nitric oxide synthase (nNOS) expression in PC12 cells. However, in the presence of nerve growth factor (NGF), nNOS expression was amplified (threefold, P < 0.05), compared to NGF alone. This increase was eliminated with pretreatment of PC12 cells with staurosporine, suggesting that the effects of ethanol on nNOS expression are mediated by a protein kinase C-dependent pathway. PMID- 10456579 TI - Zolpidem involvement on memory and hypnotic effect of ethanol in chronically ethanol-treated rats. AB - Multiple (10x) treatment of zolpidem (1.0 or 2.0 mg/kg, orally, p.o.) led to different effects in chronically ethanol-treated and control rats. In control rats, after repeated zolpidem administration, a weaker, when compared to single administration, hypnotic effect of ethanol was observed, which may be the result of tolerance developed towards the inhibitory effect of zolpidem. However, in chronically ethanol-treated rats, the multiple zolpidem treatment led to prolongation of ethanol-induced sleep similar to the values observed in non zolpidem-treated control animals. This suggests that zolpidem multiple administration may inhibit tolerance towards ethanol in chronically ethanol treated rats. In the experiment with zolpidem, there were effects on performance in a memory test and the impairment of passive avoidance task after multiple drug treatment when compared to the effects after single administration in control rats. In contrast, in chronically ethanol-treated rats, amplification of latency (especially after 2.0 mg/kg) was observed. The possible relationship between ethanol-induced sedation and latency values would be consistent with a higher contribution of the inhibitory effect of zolpidem, than a direct influence on memory processes in chronically ethanol-treated rats. PMID- 10456580 TI - Second generation effects of maternal ethanol consumption on immunity to Trichinella spiralis in female rats. AB - The deleterious effects of maternal ethanol consumption on neonatal immune development and early immune responses has been well documented. However, the effects of such neonatal exposure to maternally consumed ethanol on the neonates' immune responses in their adult life, especially in combination with additional ethanol exposure, has received little attention. For these experiments, female rats were fed on either 6% ethanol or pair-fed isocaloric control Lieber-DeCarli liquid diets for 30 days prior to, and during, pregnancy and lactation. One day after weaning their pups, the mothers were infected with 1000 Trichinella spiralis larvae, and maintained on diets for an additional 20 days. At this time, they were challenged with 2000 T. spiralis larvae, killed 3 days later, and their immune status determined. These animals served as the first generation alcohol animals. Their female offspring served as the experimental second generation animals. These animals received maternal ethanol during pregnancy and lactation and control diet during their juvenile period (from weaning to 90 days of age). They were then subjected to a schedule of ethanol or pairfeeding, identical to the first generation dams. Two groups of second generation animals were established: Group 1 was exposed to ethanol during their dam's pregnancy and lactation periods only, with no subsequent ethanol treatment; Group 2 received ethanol during their dam's pregnancy and lactation periods and then again throughout their adult experimental period. Our previous studies showed only minimal changes following a secondary challenge in T. spiralis-immunized rats; however, neonates born to alcohol-consuming mothers did show some depressed secondary immune responses when challenged soon after weaning. We chose to use a secondary immune challenge to assess further immune alterations in second generation adult animals. No differences between any of the ethanol and pair-fed groups were observed in intestinal worm burdens, which is similar to data previously reported for adult alcohol-consuming animals. However, second generation group 2 animals demonstrated significantly reduced proliferation responses to T. spiralis antigen and Concanavalin A (Con A) stimulation relative to the ethanol first generation and to the second generation Group 1 animals. This group also demonstrated significantly lower absorbencies in the ELISA assay for specific IgM and IgG anti-T. spiralis antibodies than the pair-fed, ethanol first and second generation Group 1 animals. The proportion of total T cells and cytotoxic T cells was significantly lower and the proportion of natural killer cells was elevated in both second generation ethanol Groups 1 and 2 relative to the ethanol first generation and pair-fed groups. In addition, Group 2 second generation animals showed significantly lower proportions of total leukocytes and T cells than Group 1 second generation animals. Although secondary immune responses to T. spiralis infection were not altered in rats exposed to ethanol only as adults, exposure to maternal ethanol does affect some specific immune responses in second generation adult life and maternal exposure may exert cumulative immune effects in concert with later consumption of ethanol by offspring born to alcoholic mothers. PMID- 10456582 TI - Development of alcohol deprivation effect in rats: lack of correlation with saccharin drinking and locomotor activity. AB - The present study addressed the relationship between the parameters of saccharin drinking behaviour and locomotor activity in an open field environment and long term alcohol self-administration. In a 22-day initiation phase, male Wistar rats were presented with increasing concentrations of ethanol (2-8%, v/v) in a choice with water. The rats were then given the choice between water and two ethanol solutions (8 and 16%). Every 28 days, ethanol was withdrawn for 5 days. The ethanol intake and the transient increase in ethanol consumption after each of six deprivation episodes (alcohol deprivation effect) was monitored and correlated with parameters of the subsequent saccharin drinking and open field tests. The total ethanol intake (g/kg/24 h) as well as the consumption of 16% ethanol were stable over time. However, the magnitude of the alcohol deprivation effect increased with the repeated deprivation episodes. None of the parameters measured in the open field or the saccharin drinking tests correlated with either ethanol consumption or the alcohol deprivation effect. These results suggest that (1) repeated episodes of ethanol deprivation may increase the magnitude of the alcohol deprivation effect, (2) neither saccharin drinking nor locomotor activity correlates with long-term ethanol drinking behaviour in rats. PMID- 10456581 TI - Reversal of ethanol-induced hepatic steatosis and lipid peroxidation by taurine: a study in rats. AB - Alcohol (ethanol) was administered chronically to female Sprague-Dawley rats in a nutritionally adequate, totally liquid diet for 28 days. This resulted in significant hepatic steatosis and lipid peroxidation. When taurine was administered for 2 days following alcohol withdrawal it was found to reduce alcohol-induced lipid peroxidation and completely reversed hepatic steatosis. The reversal of hepatic steatosis was demonstrated both biochemically and histologically. Two days following alcohol withdrawal, the apparent activity of the alcohol-inducible form of cytochrome P450 (CYP2E1) was unchanged although total cytochrome P450 content was increased. In addition, alcohol significantly inhibited hepatic methionine synthase activity and increased homocysteine excretion in urine. Although alcohol did not affect the urinary excretion of taurine (a non-invasive marker of liver damage), levels of serum and hepatic taurine were markedly raised in animals given taurine following their treatment with alcohol, compared to animals given taurine alone. There was evidence of slight bile duct injury in animals treated with alcohol and with alcohol followed by taurine, as indicated by raised serum alkaline phosphatase (ALP) and cholesterol. Aspartate aminotransferase (AST) was also slightly raised. The effects of taurine on reversing hepatic steatosis may be due to the enhanced secretion of hepatic triglycerides. It is suggested that increased bile flow as a result of taurine treatment may have contributed to the removal of lipid peroxides. These in-vivo findings demonstrate for the first time that hepatic steatosis and lipid peroxidation, occurring as a result of chronic alcohol consumption, can be reversed by administration of taurine to rats for 2 days. PMID- 10456583 TI - Deleterious effects of chronic moderate alcohol intake by female mice on preimplantation embryo growth in vitro. AB - The susceptibility of preimplantation stages of embryo development to preconceptional alcohol ingestion by females has had little investigation. We have recently shown that chronic 10% (w/v) ethanol intake by young female mice reduces the ovulatory response and impairs the quality of the oocytes. The aim of this study was to investigate the effects of 10% ethanol administration for 30 days on immature female mice on the day of in-vitro fertilization (day 1) and on preimplantation embryo development. Female mice were ovulated on days 27 and 29 of ethanol treatment and in-vitro fertilization was performed 16 h post-human chorionic gonadotrophin administration (day 30). The oocytes from the ethanol treated females inseminated with spermatozoa from control males, showed a significantly higher percentage of parthenogenetic activation compared to the control females. An increased percentage of fragmented oocytes was found after insemination, compared to control females. When the embryos were cultured, the percentage of 2-cell (day 2), 4-cell (day 3) embryos, and compacted morulae (day 4) was significantly reduced in treated females, compared to control females. On day 5, we found a highly significant decreased percentage of early and expanded blastocysts in the ethanol-treated females. The percentage of hatching and hatched (extruded) blastocysts was also reduced significantly in treated females at days 6 and 7 (blastocyst stages). An increased percentage of morphologically abnormal embryos was found on days 5 and 6 in ethanol-treated females compared with controls. We conclude that chronic moderate ethanol ingestion by young female mice results in decreased fertilization, embryo growth retardation, cleavage arrest, and abnormal embryo development in vitro. PMID- 10456584 TI - Intervention for excessive alcohol consumption in primary health care: attitudes and practices of English general practitioners. AB - General practitioners' (GPs') recognition of, attitudes towards, and intervention for, excessive drinking and alcohol problems among their patients were assessed in a postal questionnaire survey. Levels of recognition of, and intervention for, excessive drinking by GPs were low. GPs did not routinely enquire about alcohol and had managed only small numbers of patients specifically for excessive drinking or alcohol problems in the previous year. Enquiry about alcohol issues was elicited mainly by physical symptoms or by new patient registrations. Although 83% of GPs felt prepared to counsel excessive drinkers, only 21% felt effective in helping patients reduce consumption. Over the past 10 years, there appears to have been an increase in numbers of GPs who feel that they should be working with alcohol issues, but fewer GPs perceive themselves as being effective in this work. The main barriers to brief alcohol intervention were given as insufficient time and training, and lack of help from government policy; the main incentives related to availability of appropriate support services and proven efficacy of brief interventions. PMID- 10456585 TI - Rapid recovery from cognitive deficits in abstinent alcoholics: a controlled test retest study. AB - The pattern of cognitive deficits and their time-dependent recovery were investigated in a cohort of 49 male alcohol-dependent patients using a repeated measurement design with 49 healthy male controls matched for age, education, and marital status. We combined parts of the Halstead Reitan Battery and the Wechsler Memory Scale with tests that are widely used in German-speaking countries. Patients were tested in the first week (T1) and 5 weeks later (T2) at the end of the in-patient treatment programme. Matched controls were tested also at T1 and T2, which enabled us to take learning effects into account. At T1, the patients showed distinct cognitive deficits on 5 of 12 neuropsychological parameters (perceptual-motor speed, verbal short-term memory, verbal knowledge, non-verbal reasoning, spatial imagination). At T2, significant improvements had occurred in four of the five dysfunctional domains with a significant difference remaining in verbal short-term memory. Duration of dependency and length of abstinence prior to testing had no essential effects on neuropsychological functions. Our results provide evidence for the well-established fact that chronic alcoholism has detrimental effects on cognitive performance, but that performance improves with neuropsychological recovery which occurs rapidly within weeks when abstinence is maintained. Cognitive deficits seem to be similar across different studies and cultures. PMID- 10456586 TI - Acceptability of computerized self-report of alcohol habits: a patient perspective. AB - The acceptability of computerized assessment of alcohol habits was explored in 57 consecutive out-patients over a 6-month period. Altogether, 46 men and 11 women agreed to complete a paper and pencil questionnaire exploring their opinion about computerized assessment. The study focused on the patients' acceptance of computerized testing and also on whether some sub-groups had reservations. The participants indicated that they had no general anxiety towards computers and did not mind being assessed by their use. Nearly half of the men were not convinced of the usefulness of computers as a means of asking about alcohol habits. The same level of confidence was recorded with regard to whether doctors would make better assessments using computers. Around one-quarter of both men and women were worried that computers might cause doctors to spend less time with the patients and that staff might lose the personal contact with patients. Because of the small sample size, we conclude tentatively that a computerized lifestyle test appears to be an acceptable method both to men and women with different educational backgrounds. However, two important issues need to be further addressed, namely concerns about confidentiality and loss of personal contact. PMID- 10456587 TI - Emergency room and primary care services utilization and associated alcohol and drug use in the United States general population. AB - Heavy and problem drinking have been found to be over-represented in some studies of clinical populations, but little is known of the association of alcohol and drug use with health services utilization in the general population. General population data are reported on the association of alcohol, drug use, and demographic characteristics with use of the Emergency Room (ER) or other primary care services during the preceding year. Data analysed are from the 1995 National Alcohol Survey of respondents living in households in the 48 contiguous states. Logistic regression was used to analyse the association of substance-use variables with ER and primary care services use by gender within three ethnic groups: white (n = 1636), black (n = 1582) and Hispanic (n = 1585). Variables found to be associated with ER use were not associated with primary care services use and vice versa. Drinking and drug-use characteristics appeared to be more associated with ER use than with primary care use, and varied across ethnic and gender subgroups. The data suggest an association between treatment for alcohol or drug problems and services use, particularly among blacks and Hispanics. The data support findings from clinical studies that over-representation of substance use problems in some clinical settings may be due to the socio-demographic characteristics of those using the facility under study, and suggest that ER and other primary care service settings may be useful for identifying those with alcohol and drug-regulated problems among specific gender/ethnic groups. PMID- 10456588 TI - Regret is what you get: the effects of manipulating anticipated affect and time perspective on risky single-occasion drinking. AB - This study tested anticipated affect as a potential strategy for reducing risky single-occasion drinking (RSOD). The hypothesis was that asking respondents to focus on their anticipated affect following RSOD would lead to higher ratings of negative affect than those obtained when asking respondents to focus on their feelings towards RSOD. In turn, these negative affect ratings were hypothesized as leading to safer behavioural estimates and reductions in RSOD. The study is based on a self-report questionnaire administered at two time points. At Time 1, measures of past drinking and demographic information were collected, along with affect ratings of drinking within safer single-occasion limits and affect ratings of RSOD (within-subjects condition). Time perspective was manipulated whereby the experimental group was asked to focus on affective reactions after RSOD and the control group to focus on affective reactions towards RSOD (between-subjects condition). Two weeks later, drinking behaviour was measured. The findings showed that the time perspective manipulation resulted in significantly higher negative affect ratings in the feeling after condition than in the feeling towards condition. Further, females reported lower negative affect than males. No other main or interaction effects were found. The time perspective manipulation, however, failed to produce safer behavioural estimates and RSOD reduction at follow-up. No significant differences were found between ratings of negative affect when drinking within safe limits as compared with ratings of affect when drinking above such limits. Despite greater negative affect 'after' rather than 'toward' the target behaviour, anticipated affect following RSOD did not yield safer behavioural estimates and subsequent drinking reduction at follow-up. These findings are interpreted in the context of risk perception associated with RSOD. The implications of this study for design of interventions aimed at reducing RSOD are discussed. In particular, ways of intensifying negative affect for RSOD are considered. PMID- 10456589 TI - A comparison of cognitive performance in binge versus regular chronic alcohol misusers. AB - One hundred cases of individuals assessed for alcohol-related cognitive performance were examined. The assessment included demographic and alcohol consumption data, as well as performance on tests of auditory verbal learning, memory, motor skills, general intellectual functioning, and visuospatial functioning. All participants regularly drank in excess of 10 standard drinks/session. Fifty cases were binge drinkers who consumed alcohol on 2 days/week or less and 50 cases were individuals who consumed alcohol daily. The two groups of drinkers were statistically matched on a number of demographic and misuse factors. The results indicated similar performance for both the binge drinkers and the regular drinkers in visuo-motor speed, visuo-spatial organization and planning, learning, proactive interference, retroactive interference, and retrieval efficiency. However, performance differences were observed on tasks that required semantic organizational ability, with binge drinkers performing better than regular drinkers on these tasks. Due to the differences in the cognitive performance of the two groups, it was concluded that drinking pattern is an important factor in investigating cognitive performance in alcoholics. PMID- 10456590 TI - Preventing injuries through interventions for problem drinking: a systematic review of randomized controlled trials. AB - To assess the effect of treatment of problem drinking on injury risk, we conducted a systematic review of randomized controlled trials by searching 12 computerized databases, cross-checking bibliographies, and contacting authors and governmental agencies. We identified 19 trials of interventions for problem drinking that measured injury outcomes. Treatment for problem drinking was associated with reduced suicide attempts, domestic violence, falls, drinking related injuries, and injury hospitalizations and deaths, with reductions ranging from 27 to 65%. Interventions among convicted drunk drivers reduced motor vehicle crashes and injuries. The precision of all the point estimates was low, however. We did not combine the results quantitatively, because the interventions, patient populations, and outcomes were so heterogeneous. The results suggest that treatment for problem drinking may reduce injuries and their antecedents. Because injuries account for much of the morbidity and mortality from problem drinking, further studies are warranted to confirm these effects. PMID- 10456591 TI - Patterns of alcohol consumption in Bulgaria. AB - Alcohol is increasingly being recognized as a major cause of the greater burden of disease and premature death in eastern Europe than in the west. This requires a robust policy response but, in most countries in the region, there is little empirical data on patterns of consumption on which to base such a response. Until now, there has been no information on prevalence of drinking among the Bulgarian population. This paper describes a survey that seeks to address this weakness by describing the pattern of drinking in Bulgaria and its relationship with socio demographic factors. It uses a multivariate analysis of data on patterns of alcohol consumption from a multi-stage nationwide survey of 1550 adults in Bulgaria, in 1997 which found that overall 50.7% of men and 13.6% of women drink at least weekly. In both sexes, drinking is least common among the elderly and those living in villages. It is also less common among those reporting their financial status as poor. Muslims are less likely to drink than are orthodox Christians. Drinking is most common among those living in cities, with higher education and high incomes. Heavy drinking, defined as 80 g/day or more, is rare among women, but is ascribed to 18.2% of men. Heavy drinking was much more common among men living in Sofia and was less common among those whose financial situation was poor. At the levels of drinking reported, it can be expected that alcohol is making a substantial contribution to the burden of disease and premature mortality in Bulgaria. A coordinated, effective policy response is required. Although some of the legal prerequisites are in place, they are poorly enforced and there is no overall strategy to address this issue. PMID- 10456592 TI - Acupuncture in alcoholism treatment: a randomized out-patient study. AB - Seventy-two alcoholics were treated with acupuncture to the ear in a randomized single-blind controlled design over 10 weeks. Orthodox points and incorrect points 3-5 mm from orthodox points were used. No initial differences were found regarding social characteristics, the responses to the Swedish version of the Alcohol Use Inventory and the Three-dimensional Personality Questionnaire, indicating a successful randomization. There were non-significant tendencies towards gender differential response after acupuncture treatment (P = 0.07). There was no difference in the number of drinking days or level of craving between treatment and control patients. Among females, those in the treatment group reported reduction of anxiety after 1 month, more often than those in the control group (P < 0.05). Response to acupuncture was not related to personality or drinking pattern. Patients' experience of needle placement was similar in the study and control groups. The effects of acupuncture were less pronounced than those previously reported. PMID- 10456593 TI - A comparison of two retrospective measures of weekly alcohol consumption: diary and quantity/frequency index. AB - There is currently uncertainty regarding the relative performance of a retrospective diary (RD) and a quantity-frequency index (QFI) measure of weekly alcohol consumption. While some previous studies have found more consumption reported on an RD than a QFI, others have found the reverse. As yet, however, no study has compared computerized versions of these two measures. This cross sectional study involved administration of a computerized survey in a community based drug and alcohol treatment setting. Five hundred and eighty-six clients (420 males) attending counselling for a range of drug- and alcohol-related issues agreed to participate in the study. The major finding was that more alcohol consumption was reported on the RD, than on the QFI. Similarly, the RD detected a greater proportion of both heavy and high-risk drinkers than the QFI. It is argued that the RD may be preferable to the QFI as a measure of weekly alcohol consumption, for use in community-based treatment settings. PMID- 10456594 TI - A child's need, a parent's wish, an orthodontist's responsibility. PMID- 10456596 TI - Making decisions for children. AB - Making decisions for children is part of everyday orthodontic care. When conflicts arise between providers and parents, articulation of the ethical and legal principles guiding the process can be helpful. Parents are generally decision makers for their minor children, a presumption supported for practical and moral reasons and by legal precedent. The best interests of the child must guide both parents and providers. As the child matures, he or she should be brought into the decision making process as age and ability allow. A child's competency is a function of age, cognitive abilities, and personal experiences. The child should participate in the decision making process to facilitate trust, cooperation, and the development of future decisional capacities. The concepts of informed consent, parental permission, and child assent are compared and contrasted. Tables are provide that summarize these concepts and offer guidelines for providers working with children. PMID- 10456595 TI - Ethics for orthodontists. AB - When treating children, important ethical issues may arise relating to the best interests of the child and decision making for minors. The case of a child with a cleft lip and palate whose parents failed to bring her in for medically indicated orthodontic care is presented. Ethical features of the case are discussed, including the need to benefit the patient, avoid harm, and respect the preferences of the parents. Ethical codes of the American Dental Association and American Medical Association are referenced. Ethical dilemmas include the conflict between the orthodontist's obligation to the child and the need to respect parental autonomy. Parental autonomy is respected up until the point at which significant harm to a child may result. The orthodontist's primary ethical responsibility is to the child, not the parents. The orthodontist providing medically indicated care should involve the craniofacial team or hospital social worker when parental decision making is in question. PMID- 10456597 TI - Tooth position and speech--is there a relationship? AB - Although it is widely accepted that teeth play an important role in speech production, the relationship between tooth position and speech remains controversial. This review paper examines the relevant studies and discusses the difficulties of scientific investigation in this area. The ability of patients to adapt their speech to compensate for abnormal tooth position is recognized, but the mechanisms for this adaptation remain incompletely understood. The overall conclusion is that while certain dental irregularities show a relationship with speech disorders, this does not appear to correlate with the severity of the malocclusion. There is no definitive proof that alteration of tooth position can improve articulation disorders. PMID- 10456598 TI - A cephalometric study of class II Division 1 malocclusions treated with the Jasper Jumper appliance. AB - This lateral cephalometric study investigated the dental and skeletal effects of the Jasper Jumper appliance used in the correction of Class II Division 1 malocclusions. A sample of 36 growing patients treated with the Jasper Jumper appliance was divided into two groups: (1) 24 patients with records obtained at the start and completion of orthodontic treatment, and (2) 12 patients with records available at the beginning and end of the Jumper phase of treatment. Treatment effects were determined by statistical comparisons of cephalometric changes in the patients relative to age-adjusted cephalometric standards, and from structural superimpositions. While the Jumpers were in place, maxillary incisors were retroclined and the molars were moved distally, tipped back, and intruded. The mandibular incisors were proclined and intruded, while the molars were translated mesially, tipped forward, and extruded. Skeletal measures showed reduced forward maxillary displacement and no significant alteration of horizontal mandibular growth. During orthodontic finishing, molar tipping and maxillary incisor retroclination were corrected, although the mandibular incisors remained proclined. In summary, this study found that the Jasper Jumper appliance corrected Class II discrepancies largely through maxillary and mandibular dentoalveolar effects and, to a limited extent, by restraint of forward maxillary growth. PMID- 10456599 TI - Class II Division 2 malocclusion: genetics or environment? A case report of monozygotic twins. AB - A pair of monozygotic twins with different malocclusion phenotypes (Class II Division 2 and Class II Division 1) is presented. The case report supports the hypothesis that heredity is not the sole controlling factor in the etiology of Class II Division 2 malocclusion. PMID- 10456600 TI - Evaluation of orthodontists' perception of treatment need and the peer assessment rating (PAR) index. AB - This paper examines the relationship between orthodontists' subjective assessment of treatment need and objective measurements obtained during standardized intra- and extraoral examinations. Logistic regression modeling was used to develop predictive models of treatment need. Data were obtained from 1155 eighth-grade students by four orthodontists who used standardized examination forms to assess demographics, trauma, skeletal relationships, morphologic malocclusion traits, and mandibular function. At the conclusion of the examination, the orthodontist rated the subjective treatment need as none, elective, recommended, soon, or immediate. For some analyses, the categories were collapsed to represent no need and need. The peer assessment rating (PAR) index (American validated version) was computed from the clinical exam findings and scoring of dental models; PAR scores were used to document malocclusion severity and treatment difficulty. Spearman rank correlation coefficients quantified the relationship between PAR scores and need categories. Logistic regression analysis modeled treatment need using components of the PAR index as well as other variables. The components of these models, as well as sensitivity and specificity, were compared with malocclusion severity/treatment difficulty scores obtained from malocclusion assessments using the PAR index. The five subjective treatment need categories and the PAR index scores were significantly correlated (rho = 0.62, p<0.001). Significant differences were detected between the need and no need groups for all PAR components (p<0.001). PAR index scores and predicted probabilities from logistic regression models performed equally well for classification purposes (no need, need). The data suggest that the PAR index is highly correlated with orthodontists' subjective assessment of treatment need when that assessment is made in the absence of financial considerations and patient desires. PMID- 10456601 TI - Morphology and growth in convex profile facial patterns: a longitudinal study. AB - Two groups of females, one with normal anteroposterior jaw-base relationships and the other with distal jaw-base relationships, were selected at age 6 and compared longitudinally up to age 18. The purposes of this study were, first, to reveal morphological factors that caused or contributed to a distal jaw-base relationship, and second, to compare growth in the two groups. A distal jaw-base relationship is not a morphological entity caused by some specific aberration in the cranial base or jaws. Rather, it is the result of a combination of predisposing deviations with varying degrees of gravity. A short mandibular corpus and a large MP-SN angle were the only deviations with significant group differences. Distal jaw-base relationships generally worsened with age as compared with normal anteroposterior jaw-base relationships. Inadequate increase in mandibular corpus length in the 6- to 12-year period contributed to the worsening, as did the mandible growing more vertically than normal after age 12. PMID- 10456602 TI - A technique for three-dimensional cephalometric analysis as an aid in evaluating changes in the craniofacial skeleton. AB - A technique is presented for creating a standardized view for use in three dimensional cephalometry. Three-dimensional cephalometric analysis has become a valuable tool in the assessment of skeletal remodeling, contour changes, and changes in proportion that occur with aging. Computer-assisted tomographic data can be used in either coronal or axial sections; the present study was performed with axially acquired data that was processed in several steps to achieve a standardized position of the facial skeleton for further evaluation. This technique may be a useful adjunct to standard cephalometrics to evaluate the craniofacial skeleton. PMID- 10456603 TI - Very early face mask therapy in Class III children. AB - The purpose of this study was to examine the effects of very early face mask therapy in children with Class III malocclusion. At pretreatment (T0), 40 female subjects who were eventually treated showed a more severe Class III pattern (ANB 0.1 degrees) than did 28 skeletal Class III female subjects who remained untreated (ANB 0.4 degrees), as observed in the Wits analysis. Posttreatment results (T1) showed significant (p<0.01) anterior advancement of the maxillary components, backward rotation of the mandible without increased lower anterior facial height, and an improved incisor relationship. Comparison of posttreatment (T1) and postretention (T2) records, however, revealed no increase in SNA in the treated group. SNA did increase in the untreated group, with no significant difference in ANB angle. The x-components of B-point and Me showed a significant (p<0.05) difference between the two groups. At postretention (T2), N-S-Ba, N-S Ar, and CC-Ba tended to increase more in the treated group than in the untreated group. PMID- 10456604 TI - Influences of vertical occlusal discrepancies on condylar responses and craniofacial growth in growing rats. AB - The present study was conducted to investigate the influence of vertical occlusal discrepancies on condylar remodeling and the subsequent craniofacial growth in growing rats. Thirty 4-week-old male Wistar strain rats were used. A 1-mm-thick metal plate was bonded onto the occlusal surface of the maxillary molars to increase posterior dentoalveolar height. During the early phase of the experiment, the thickness of the proliferative and maturative/hypertrophic zones in the anterior and superior portions of the condyle was significantly smaller in the experimental group than in the controls. The number of TRAP-positive cells was significantly greater in the experimental group than in the controls. At the end of the experiment, decreased ramus height and a large gonial angle were found in the experimental group. Changes in the intra-articular environment associated with vertical occlusal discrepancies may influence condylar and craniofacial growth in growing individuals, although some adaptive response of the condyle may be induced if growth potential remains. PMID- 10456605 TI - Influence of dietary n-3 polyunsaturated fatty acid on experimental tooth movement in rats. AB - This study was conducted to investigate the influence of dietary n-3 polyunsaturated fatty acid on experimental tooth movement. This acid substantially reduces the production of arachidonic acid. Sixty 4-week-old male Wistar strain rats were divided into experimental and control groups. Animals in the experimental group were fed a purified diet containing 10% refined fish oil (rich in n-3 fatty acid); control animals were fed a diet containing 10% corn oil (rich in n-6 fatty acid). After 6 weeks, the maxillary first molars were moved buccally with an initial force of 20 g for periods of 0, 3, 7, or 14 days. Tooth movement in the experimental group was 80% of that seen in the controls. The number of osteoclasts on the pressure side during tooth movement was nearly 60% of that seen in controls, and the degree of bone resorption was 80%. The data suggest that a diet enriched with fish oil reduces osteoclastic activity and subsequent alveolar bone resorption that is the key to experimental tooth movement. PMID- 10456606 TI - Displacement and stress distribution in the temporomandibular joint during clenching. AB - The aim of this study was to analyze biomechanical reactions in the mandible and TMJ during clenching under various restraint conditions. A three-dimensional finite element model of the mandible, including the TMJ, was created for test purposes. The results were as follows: (1) Under any restraint conditions, displacement was greatest on the surface of the condyle and less on the articular disc and the surface of the glenoid fossa, in that order. Resultant stresses followed the same trend. (2) Displacement and stress were greatest when the lower central incisor was restrained and attenuated as the posterior teeth were restrained. Because the biomechanical reaction of the TMJ during clenching was greatest when the lower central incisor was restrained, premature contact of these teeth may be one of the factors involved in the initiation of temporomandibular arthrosis. PMID- 10456607 TI - Molarization of the lower second premolars. AB - This paper presents a case of extreme tooth variation. The patient was first observed during the mixed dentition period, when she presented a mild Class II malocclusion with increased overjet and acceptable overbite. In a panoramic radiograph, the presence of lower second premolars of disproportionate dimensions was discovered. When these oversized premolars erupted, the Class I malocclusion tended toward Class III, with an edge-to-edge bite. This created an unstable occlusion and the possible need for extractions. PMID- 10456608 TI - Antimicrobial resistance in gram-positive bacteria: the myth of the MIC. Minimum inhibitory concentration. AB - Although much progress has occurred in the fight to treat infection, resistant bacteria increasingly are becoming problematic. The minimum inhibitory concentration is easily determined, yet its definition, clinical application, and relevance are confusing. In addition, errors can occur during in vitro susceptibility testing. The mechanism of resistance for different organisms is discussed as are the pitfalls of susceptibility testing and the prescription of antimicrobial agents. PMID- 10456609 TI - Successful interventions for gram-negative resistance to extended-spectrum beta lactam antiobiotics. AB - Antibiotic resistance among nosocomial pathogens in this country's hospitals adds significantly to patient morbidity and mortality, and the cost of health care. Optimism for identifying antimicrobial agents that would "solve the problem" of resistance has been replaced by a much more guarded and realistic view of the battle between humans and pathogenic microorganisms. Efforts now are more appropriately directed toward limiting, rather than completely eliminating, resistance, generally by either infection control or antibiotic control measures, and sometime combinations of the two. Methicillin-oxacillin resistance in Staphylococcus aureus (MRSA) results from the expression of an acquired penicillin-binding protein (PBP 2a) that is not transferable in vitro. In most hospitals, even those with high percentages of MRSA, relatively few resistant clones are identified, suggesting transmission of individual strains throughout the hospital population. Because person-to-person spread is so important in transmission of MRSA, strategies aimed at preventing transmission of the resistant strains are remarkably effective when strictly enforced. Ceftazidime resistance in Enterobacteriaceae results from point mutations within genes that encode widely prevalent and often transferable plasmid-mediated enzymes. In addition, mutations of these genes that allow hydrolysis of cephalosporins usually result in decreased activity against other drugs, including the penicillins and beta-lactamase inhibitors. Effective measures to control ceftazidime-resistant Enterobacteriaceae have as their cornerstone limiting administration of antibiotics that select for the emergence and spread of these mutations, especially ceftazidime. The importance of infection-control techniques in limiting the prevalence of ceftazidime-resistant Enterobacteriaceae is less well established. Methods that are informed by a detailed understanding of the molecular mechanisms of resistance and resistance spread offer the best hope for limiting dissemination of antibiotic-resistant bacteria in a cost-effective manner. PMID- 10456610 TI - Decreased antimicrobial resistance after changes in antibiotic use. AB - Vancomycin-resistant enterococci (VRE) and methicillin-oxacillin-resistant Staphylococcus aureus (MRSA) originally predominated in large medical centers; however, these isolates are now common in community hospitals and community clinics. No simple answer is available regarding control of antimicrobial resistant bacteria, especially VRE and MRSA, as their numbers increase and pose a more serious threat to patient care. The source of colonization is often difficult to identify because of transport of patients among different locations on the continuum of care (e.g., hospital to extended care facility to home and back). At one hospital, control strategies greatly reduced the occurrence of gram negative bacteria such as VRE. Since 1994, VRE declined from 16% to 5%. Similarly, the number of MRSA isolates declined from 35% to 23%. These declines are attributed to a cohesive working relationship among pharmacists, microbiologists, and infectious disease physicians and personnel, and to a decision to decrease administration of cephalosporins in favor of piperacillin tazobactam. PMID- 10456611 TI - Clinically significant differences in the International Normalized Ratio measured with reagents of different sensitivities. SPAF Investigators. Stroke Prevention in Atrial Fibrillation. AB - The International Normalized Ratio (INR) system was introduced a decade ago as a way of standardizing the results of prothrombin time testing for patients taking oral anticoagulants. A strong emphasis has been placed upon using thromboplastin reagents that are very sensitive to the effects of oral anticoagulants upon the prothrombin time [i.e. reagents with low International Sensitivity Index (ISI)]. In order to assess how well the INR system functions as currently used in clinical laboratories, we compared the INRs determined using thromboplastins of differing ISIs in samples collected during a large clinical trial of oral anticoagulation for atrial fibrillation (Stroke Prevention in Atrial Fibrillation III trial). Frozen plasma was subjected to prothrombin time testing using thromboplastins with ISIs ranging from 0.97 to 2.49. INRs were calculated using machine-specific ISIs and Westgard's rules were followed to maintain quality control. An unanticipated coagulometer failure allowed a determination of the effect of machine recalibration upon the INR of control plasmas. The correlation between each pair of INRs obtained from 1181 plasmas was high (> 0.9), but the differences between reagents were statistically different from zero (P<0.001 for pairwise comparisons). Plasmas had INRs within the therapeutic range (2.0-3.0) with one reagent but not with another in an average of 20% of instances. Among the 20% discordant pairings, 43% (8.5% of the total tested) showed a difference in INR of more than 0.2 INR units above or below the target range. Low ISI thromboplastins did not perform better in this pairwise comparison than other reagents or the locally determined INR. Recalibration of a coagulometer resulted in a significant change in the INRs obtained from control plasmas (P<0.0001), which confirms and extends the observations of other authors concerning the sensitivity of the INR to coagulometer-related variables. There was a clinically significant difference in the INRs obtained with different thromboplastins, and low ISI reagents did not perform better than others. Since the risk of thrombosis rises sharply below the lower limit of the currently recommended target ranges, consideration should be given to narrowing the recommended range, or advising clinicians to aim for its mid-point. These findings illustrate the difficulties in imposing standardization upon coagulation testing after a test is in widespread use. PMID- 10456612 TI - Detection of two novel mutations (nt2762, exon 2, CAG to TAG, and nt2483 or 2484, exon 2, +A) in individuals with congenital type I antithrombin deficiencies. AB - Genetic analyses were performed on two Japanese kindreds with congenital type I antithrombin deficiency causing recurrent thrombosis. The seven exons and flanking intron regions of the antithrombin gene were amplified by polymerase chain reaction followed by direct nucleotide sequencing. In case 1, one novel nonsense mutation, CAG to TAG at codon 100, nucleotide 2762, in exon 2, was detected. In case 2, one novel minor insertion, AT to AAT at codon 7, nucleotide 2483 or 2484, in exon 2, was detected, leading to a frameshift which resulted in a stop codon TGA at codon 32. PMID- 10456613 TI - Thrombopoietin and thrombospondin in renal allograft recipients. AB - Recipients of renal transplants appear to be at increased risk of thromboembolic events. Despite accumulating evidence for the hyperreactivity of platelets, the primary regulator of thrombopoiesis, thrombopoietin (TPO), has not yet been studied in renal transplant recipients. Thus, the aim of the present study was to quantify the levels of TPO and to assess its contribution to increased platelet reactivity in recipients of renal allografts. Serum concentrations of thrombospondin (TSP) were also determined in patients undergoing renal transplants in order to evaluate the role of this multifunctional protein in platelet hyperaggregability. Serum levels of TPO were significantly lower in renal transplant recipients (n = 27) than in healthy controls (30.8+/-20.6 pg/ml versus 129.9+/-113.6 pg/ml, P = 0.001). Serum concentrations of TPO were correlated neither with serum levels of creatinine nor duration of transplantation. However, levels of TPO were negatively correlated with platelet counts (r = -0.50, P = 0.007) in recipients of renal transplants. Plasma levels of TSP were higher in renal transplant patients than in the control group (104.5+/-54.7 ng/ml versus 63.4+/-41.5 ng/ml, P = 0.003). No significant correlation was found between levels of TPO and TSP. We conclude that, rather than the allograft function, the platelet mass determines the levels of TPO in recipients of renal transplants. Despite the low serum levels of TPO, and increased concentrations of TSP, TPO might still play a role in the hyperaggregability of platelets in patients undergoing renal transplants. PMID- 10456614 TI - Effect of ciprofibrate on fibrinogen synthesis in vitro on hepatoma cells and in vivo in genetically obese Zucker rats. AB - In this study, we have shown that oncostatin M and interleukin-6 induce a dose- and time-dependent increase in fibrinogen levels in the conditioned medium of human hepatoma cells (HepG2). When HepG2 cells were treated simultaneously with oncostatin M or interleukin-6 and ciprofibrate (100 nmol/l), the production of fibrinogen in the conditioned media was strongly affected and a significant decrease in the mRNA levels of the fibrinogen beta chain was observed. Oncostatin M- and interleukin-6-induced fibrinogen release was inhibited in a dose-dependent manner by ciprofibrate and, to lesser extent, by bezafibrate, fenofibric acid and clofibric acid. In vivo, increased plasma and platelet levels of fibrinogen were observed in genetically obese Zucker rats (fa/fa) compared with Zucker lean (fa/ ) rats. In these rats, a 14-day oral treatment with ciprofibrate (10 mg/kg, per. os.) induced a statistically significant decrease (P > 0.05) in plasma concentrations of total cholesterol and triglyceride but also in plasma and platelet levels of fibrinogen. In order to determine the consequences of such an effect on fibrinogen, the ability of ciprofibrate to affect venous stasis was determined in a stasis-induced venous thrombosis model in Zucker rats. Under low thrombogenic challenge, ciprofibrate significantly inhibited thrombus formation (67+/-12%, P > 0.05), demonstrating for the first time that a potent hypolipemic compound exhibits an antithrombotic effect. PMID- 10456615 TI - Failure of soluble fibrin polymers in the diagnosis of clinically suspected deep venous thrombosis. AB - A new diagnostic test has recently become available which is highly specific for plasma soluble fibrin polymers, the thrombus precursor protein (TpP) enzyme immunoassay. In order to evaluate the diagnostic accuracy of this test and that of a new rapid and automated test for the determination of D-dimers, the BC D dimer test, in patients with clinically suspected deep vein thrombosis (DVT), 70 consecutive symptomatic patients underwent laboratory analysis with both tests and with the classic enzyme-linked immunosorbent assay (ELISA) D-dimer test, followed by the execution of a compression ultrasound (CUS) test of the affected limb. Patients with a positive CUS test were considered to have DVT (20 of 70), whereas those with a serially negative test and an uneventful 3-month follow-up test were regarded as not having DVT (50 of 70). The sensitivity of the TpP test (45.0%) was significantly lower than that of both the BC D-dimer test (80.0%; P = 0.02) and the classic ELISA test (90.0%; P = 0.002). The specificity of the TpP test (66.0%) did not differ from that of either D-dimer test (60.0 and 64.0%, respectively). The negative predictive value of the TpP test (75.0%) was significantly lower than that of the classic ELISA D-dimer test (94.1%; P = 0.02), which in turn did not differ from that of the BC D-dimer test (88.2%). The positive predictive value was similarly low for each investigated test (34.6, 44.4, and 50.0%, respectively). In conclusion, the TpP test can neither be used to detect a DVT nor to exclude its development in patients with the clinical suspicion of this disease. By contrast, the BC D-dimer might safely replace the classic ELISA test for excluding DVT in symptomatic patients. PMID- 10456616 TI - Thrombophilic predisposition in stroke and venous thromboembolism in Danish patients. AB - The present study describes 403 patients with thrombosis, from a uniform ethnic and geographical background. Two-hundred-and-seven individuals had suffered mild or moderate stroke and 196 individuals suffered venous thromboembolism. We recorded levels of antithrombin, protein C and protein S, plasminogen and plasma homocysteine, and the presence of the factor V Leiden mutation, the prothrombin 20210G-->A variant, and the methylenetetrahydrofolate reductase (MTHFR) 677C-->T polymorphism. Controls for the mutation frequencies consisted of Guthrie card blood spots from a cohort of new-born babies. The cumulative prevalence of deficiencies in antithrombin, protein C, protein S or plasminogen was 2.4% in patients with stroke and 11.2% in patients with venous thrombosis. The factor V Leiden mutation was present in 11.1% of patients with stroke and 26.5% of patients with venous thrombosis, compared with 6.6% of controls (n = 4188; P < 0.05 and P < 0.0001, respectively). The prevalence of the prothrombin 20210A variant was 3.1% in patients with venous thrombosis, 1.9% in patients with stroke and 2.0% in controls (n = 500; P > 0.05). Hyperhomocysteinemia was present in 16.0% of patients with stroke and 17.6% of patients with venous thrombosis. The prevalence of the MTHFR 677T/T genotype was no different in patients with stroke (10.6%) and venous thrombosis (8.7%) than in controls (8.3%; n = 1084; P > 0.05); thus, it apparently contributed to thrombosis only via its influence on total plasma homocysteine, which was significantly increased in patients with the T/T genotype (P < 0.001). The MTHFR T/T genotype did not further increase the risk for thrombosis in carriers of the factor V Leiden mutation. Overall, thrombotic events were associated with a known risk factor in 27% of patients with stroke and 55% of patients with venous thrombosis. PMID- 10456617 TI - Alterations to the fibrinolytic enzyme system in patients with non-small cell lung carcinoma. AB - Although fibrinolysis has been implicated in the progression and metastasis of lung cancer, no detailed study has been carried out on components measured in samples from both plasma and tumour. This study thus provides the first comprehensive data obtained from 166 patients diagnosed with non-small cell lung carcinoma. Plasma samples were obtained at diagnosis and tumour samples during surgical resection. Appropriate control samples were obtained from normal subjects and patients with chronic obstructive airways disease (plasma) and from organ donors (normal lung tissue). Assays were performed on plasma and tissue extracts for tissue plasminogen activator, urokinase-like activator and plasminogen activator inhibitor (activity and antigen in all cases), together with plasmin-antiplasmin complex, soluble fibrin, D-dimer and thrombin antithrombin complex. Levels of D-dimer, thrombin-antithrombin complex and plasmin-antiplasmin complex were all significantly higher in plasma from patients, whereas urokinase-like activator activity was reduced. Only two parameters were significantly altered in both the core and periphery of tumour tissue: levels of D-dimer were increased and tissue-type plasminogen activator activity was reduced. Interestingly, significant differences in levels of other fibrinolytic parameters were detected in the core and periphery of tumours. Significant activation of fibrinolysis was indicated in patients, although the origin of this could not be related consistently to changes in levels of plasminogen activator and inhibitor. PMID- 10456618 TI - Lipid-lowering therapy corrects endothelial cell dysfunction in a short time but does not affect hypercoagulable state even after long-term use in hyperlipidemic patients. AB - Lipid-lowering therapy reduces cardiac events to an extent that is disproportionate to the small degree of regression of coronary atherosclerosis observed among hyperlipidemic patients. We prospectively investigated the effects of lipid reduction using simvastatin on the endothelial dysfunction and hypercoagulability found in hyperlipidemic patients. We measured levels of coagulation factors [factor VII (FVII) coagulant activity (FVIIc), FVII antigen (FVIIAg), activated FVII (FVIIa), and fibrinogen], and markers of coagulation activation [prothrombin fragment 1 + 2 (F1 + 2)] and endothelial cell dysfunction [von Willebrand factor (vWF)] in 20 hyperlipidemic patients, 20 hypertensive patients, and 20 normotensive normolipidemic controls. The levels of FVIIa, FVIIc, FVIIAg, F1 + 2, and vWF were all higher in hyperlipidemic patients, but only FVIIa, F1 + 2, and vWF levels were higher in hypertensive patients than in controls. We measured the above parameters in 13 hyperlipidemic patients before and after 1, 3, 6, 12 and 24 months of simvastatin therapy and compared these values with those in 15 hypertensive patients at baseline and after 12 and 24 months. The median (25th-75th percentile) level of total cholesterol was decreased from 259 (255-278) to 206 (176-220) mg/dl after 1 month of simvastatin therapy and this reduction persisted for 2 years. The plasma level of vWF [136% (113-158%)] was not changed after 1 month of administration of simvastatin [132% (115-153%)], but was decreased after 3 months of treatment [114% (96-128%), P<0.01]. This decrease also persisted for 2 years during simvastatin therapy and both of these reductions were significant, compared with levels in hypertensive patients. In contrast, levels of fibrinogen, FVIIc, FVIIAg, FVIIa, and F1 + 2 did not change throughout the 2 years of simvastatin therapy. We conclude that lipid reduction using simvastatin corrects endothelial cell dysfunction but not hypercoagulability in hyperlipidemic patients. The improvement in endothelial cell function brought about by lipid-lowering therapy might contribute to the reduction in cardiac events within a relatively short time period in hyperlipidemic patients. PMID- 10456619 TI - Soluble intercelluar adhesion molecule-1, E-selectin, vascular cell adhesion molecule-1 and von Willebrand factor in stroke. AB - We investigated the role of endothelial cell and leukocyte adhesion in the pathophysiology of acute stroke. The immunocytochemical expression of adhesion molecules in brain tissue from six patients who died following acute ischaemic stroke showed weak endothelial expression of intercellular adhesion molecule-1 (ICAM-1), but intense expression of vascular cell adhesion molecule-1 (VCAM-1) by astrocytes and endothelial cells from the infarcted, but not the non-infarcted areas. We also measured soluble adhesion molecules E-selectin, ICAM-1, VCAM-1 and von Willebrand factor (all by enzyme-linked immunosorbent assay) in 21 patients after an acute ischaemic stroke (ictus < 12 h), and again 3 months later. Blood levels in the stroke patients were compared with 82 healthy controls and 22 subjects with carotid atherosclerosis. Compared with healthy controls, both patient groups had raised levels of von Willebrand factor (P < 0.02) but the level of soluble VCAM-1 was raised only in patients with acute stroke (P < 0.02). Levels of von Willebrand factor and soluble VCAM-1 in the stroke patients were still high at 3 months follow-up. We suggest that there might be changes in adhesion molecule expression and release in the acute and chronic stages of ischaemic stroke, where blood levels are related to immunocytochemical findings on infarcted brain. These changes may perhaps be part of the complex pathophysiological responses to infarction and repair of brain tissue following stroke. PMID- 10456620 TI - Assessment of coagulation system activation using spot urine measurements. AB - Coagulation system activation is most commonly assessed by measuring levels of one or more proteins in peripheral blood. Because faulty blood-drawing can cause activation of the coagulation system, artifactual elevations of such markers have been reported. We have therefore investigated the possibility of using randomly collected ('spot') urine samples as a non-invasive means of assessing the state of coagulation system activation. Using a commercially available enzyme-linked immunosorbent assay kit designed to measure plasma levels of fragment 1 + 2, we found immunoreactive fragment 2 in healthy control subjects, and significantly increased levels in diabetic and non-diabetic pregnant subjects, and patients with venous thromboembolism, prostate cancer, and diabetes. Measurements of excretion of immunoreactive fragment 2 are worth further study as an adjunct or alternative to plasma-based assays designed to detect or quantify coagulation system activation. PMID- 10456621 TI - Withdrawal of warfarin after deep vein thrombosis: effects of a low fixed dose on rebound thrombin generation. AB - The optimal procedure for withdrawal of warfarin in patients with deep vein thrombosis (DVT) is still not defined. Rebound thrombin generation, which occurs after the withdrawal of warfarin, has been said to be associated with early recurrence of DVT. The aim of this study was to compare two procedures for warfarin withdrawal after the first episode of DVT with respect to rebound thrombin generation. Forty-one consecutive patients were randomly assigned to abrupt withdrawal of warfarin (group A), or to an additional month of warfarin, at the fixed dose of 1.25 mg/day (group B). Plasma samples were withdrawn for the assay of prothrombin fragment F1+2 (F1+2), protein C, factor VII and International Normalized Ratio (INR), before any anticoagulant treatment (I), during initial heparin (II) and full dose warfarin (III), at the end of full dose warfarin (IV) and then 1 (V), 4 (VI), 5 (VII) and 9 (VIII) weeks after randomization. The mean duration of full-dose warfarin treatment, the mean warfarin dose and the mean INR during full-dose warfarin treatment were similar in the two groups. A decrease in F1+2 was observed during heparin and warfarin treatment (II, 1.7 nmol/ml; III, 1.0 nmol/ml; IV, 0.7 nmol/ml; versus I, 3.5 nmol/ml; P<0.01). After the withdrawal of warfarin, an increase in F1+2 was observed in both groups, but without significant statistical differences (group A: V, 1.2 nmol/ml; VI, 1.5 nmol/ml; VII, 1.6 nmol/ml; VIII, 1.1 nmol/ml; group B: V, 1.3 nmol/ml; IV, 1.5 nmol/ml; VII, 1.4 nmol/ml; VIII, 1.4 nmol/ml). No significant difference between the two groups was observed in the recovery of protein C and factor VII. Four patients experienced a recurrence of DVT, three in group B and one in group A. Our findings confirm that rebound thrombin generation occurs in patients with DVT after the withdrawal of warfarin. Rebound thrombin generation is not reduced by a low, fixed dose of warfarin. PMID- 10456622 TI - Venous thromboembolism at a young age in a brother and sister with coinheritance of homozygous 20210A/A prothrombin mutation and heterozygous 1691G/A factor V Leiden mutation. AB - We report on members of a Turkish thrombophilic family with coinheritance of the prothrombin mutation PT20210A and the factor V Leiden mutation. The 23-year-old propositus and his elder sister both had episodes of venous theomboembolism at a young age (23 years and 26 years, respectively) and are homozygous for the PT20210A mutation and heterozygous for the factor V Leiden mutation. The 51-year old father is suffering from coronary heart disease and is heterozygous for both thrombophilic mutations. The asymptomatic 43-year-old mother is heterozygous for the PT20210A mutation, but without activated protein C resistance. Two other children, a 20-year-old girl who is homozygous for the PT20210A mutation and a 13 year-old boy who is heterozygous for the PT20210A mutation, are both free from activated protein C resistance and thrombosis. This report provides further evidence for an early onset of thromboembolic disorders in individuals with an homozygous state of the prothrombin variant 20210A/A and coinheritance of another thrombophilic mutation. Consensus guidelines are required for the treatment and prophylaxis of patients and subjects who remain asymptomatic with homozygous or more than one heterozygous genetic defect associated with thrombophilia. PMID- 10456623 TI - The O blood group protects against venous thromboembolism in individuals with the factor V Leiden but not the prothrombin (factor II G20210A) mutation. AB - We investigated the influence of the ABO blood group in the observed prevalences of the recently described factor V R506Q and factor II G20210A mutations in a thrombotic population. We determined the ABO blood group in a sample of 178 unselected patients (aged 17-83 years), diagnosed at our center with deep vein thrombosis or pulmonary embolism. The results of this study show a high prevalence of thrombosis in the non-O blood group. In the general population, the prevalence as a fraction of the O blood group was 2.69 (confidence interval 1.90 3.82). Of the factor V R506Q carriers (n = 28), only one had O group blood and 27 of 28 were non-O (24 A, one B and two AB). However, within the group of factor II G20210A carriers (n = 17), seven had O, nine A and one B type blood. The prevalence of the factor V R506Q mutation within the O blood group was 2.4% (one of 42), significantly lower than in the A group (23.3%, 24 of 103; P = 0.002), or in the overall non-O group (19.9%, 27 of 136; P = 0.006). This prevalence was similar to that observed previously in the non-thrombotic population in our area (3.5%; P = 0.9). We analyzed the clotting activity of factor VIII and we found higher levels in the non-O group (1.78+/-0.61 U/ml) than in the O blood group (1.30+/-0.51 U/ ml; P < 0.0001). We speculate that factor Va in individuals with the factor V Leiden mutation could interact with the high levels of factor VIII clotting activity as a necessary cofactor. PMID- 10456624 TI - Implantable defibrillators and electromagnetic interference. PMID- 10456625 TI - Implantable cardioverter defibrillators and cellular telephones: is there any interference? AB - The aim of our study was to consider cellular telephone interference using different cellular telephones and implantable cardioverter defibrillator (ICD) models. Thirty (26 men, 4 women) patients with ICDs were considered during follow up. The ICD models were: Telectronics (7), CPI (7), Medtronic (7), Ventritex (5), and Ela Medical (4). All patients were monitored with surface ECG; permanent telemetric endo-ECG monitoring was activated. Then, the effect of two different European telephone systems were tested: TACS system (Sony CM-R111, 2W power) and GSM system (Motorola MG1-4A11, 2 W power). For both systems, the effect during call, reception, active conversation (dialogue), and passive conversation (listening) were observed. Cellular telephones were located first in contact with the programming head, then near the leads system, and lastly, in the hands of the patient. At the end of the evaluations, memories were interrogated again to check for false arrhythmia detections. In five of these patients during arrhythmia induction at device implant (first implant or ICD replacement), we also evaluated possible interference between cellular telephones in the reception phase and the ventricular fibrillation detection phase of the ICD. All evaluated models showed significant noise in the telemetric transmission when the cellular telephone (both TACS and GSM) was located near the ICD and the programming head; noise was particularly significant during call and reception, in most cases leading to loss of telemetry. No false arrhythmia detections have been observed during tests with cellular telephones located on the ICDs. During tests performed with cellular telephones located near the leads or in the hands of patients, no telemetric noises orfalse arrhythmia detections were observed. During induced ventricularfibrillation and cellular telephones in reception mode near the device, the arrhythmia recognition was always correct and not delayed. In conclusion, present ICD models seem to be well protected from electromagnetic interference caused by European cellular telephones (TACS and GSM), without under /oversensing of ventricular arrhythmias. However, cellular telephones disturb telemetry when located near the programming head. ICD patients should not be advised against the use of cellular telephones, but it has to be avoided during ICD interrogation and programming. PMID- 10456626 TI - A randomized controlled clinical trial comparing ventricular fibrillation detection time between two transvenous defibrillator models. The Acute Ventak AV investigator group. AB - The Ventak AV is an implantable cardioverter defibrillator with dual chamber pacing capability. Features include detection and treatment of ventricular arrhythmias, detection of atrial arrhythmias, as well as dual chamber pacing. The objective of the investigation was to verify the efficacy of the Ventak AV in detecting ventricular fibrillation in the presence of dual chamber pacing. Thirty three patients, who were to receive an implantable defibrillator were randomized (1:1) in a paired comparison study to the Ventak AV (study device) and the Ventak Mini (control) during defibrillation threshold testing. In order to create a "worst case scenario" for sensing of ventricular fibrillation, pacing was performed at high lower rate limit values (Ventak AV DDD pacing at 150/min, Ventak Mini at VVI 100/min). Ventricularfibrillation was induced and the randomized device was allowed to detect and treat the arrhythmia. This test was repeated for each patient using the alternate device in a randomized order, such that all patients were tested with both devices. The mean ventricular fibrillation detection time for the Ventak AV was 2.0+/-0.11 seconds and for the control device the detection time was 1.8+/-0.11 seconds (P = 0.26). Appropriate tachyarrhythmia therapy decision was documented in all episodes for both devices. The study patient population demonstrated equivalent ventricular fibrillation detection time between the Ventak AV and the Ventak Mini. The Ventak AV demonstrated effectiveness in detecting ventricular fibrillation in the presence of high rate dual chamber pacing. PMID- 10456627 TI - Dual chamber pacing for neurally mediated syncope with a prominent cardioinhibitory component. AB - The role of cardiac pacing for treatment of recurrent neurally mediated syncope (NMS) remains controversial. We hypothesized that dual chamber pacing in NMS patients with a prominent cardioinhibitory component may be beneficial. Twelve patients (mean age = 37.8+/-17 years, range 15-78 years, 7 men and 5 women) with a mean of 4+/-2.2 episodes of syncope underwent tilt table evaluation. Patients were passively tilted to 70 degrees head-up position for 20 minutes and then returned to the supine position. Isoproterenol was then infused at 1-2 microg/min to increase heart rate by > or = 25% and tilt was repeated. Patients lost consciousness after 16+/-6 minutes of tilt; nine patients had syncope in the baseline state and three during isoproterenol infusion. All patients had at least 5 seconds of asystole with a mean of 9.5+/-4 seconds (range 5-20 s). A dual chamber permanent pacemaker with a special feature allowing heart rate acceleration in response to bradycardia was implanted in all patients. During a mean follow-up of 18.6+/-4.2 months, 11 (92%) of these patients were free of syncope and had negative tilt table test. One (8%) patient had two episodes of syncope. We conclude that dual chamber pacing may be beneficial in patients with NMS with a prominent cardioinhibitory component. PMID- 10456628 TI - A "dysautonomic" head-up tilt test pattern in elderly patients with neurocardiogenic syncope. AB - The characteristics of neurocardiogenic syncope (NCS) in elderly patients remain unclear. We compared the hemodynamic profiles of young and older patients with consecutive and positive head-up tilt tests (HUT). Continuous, noninvasive, and reliable monitoring of arterial pressure (AP) and heart rate (HR) was done throughout 46 consecutive positive HUTs of symptomatic patients. The population (12-82 years old) was divided into two groups: younger patients, Y (n = 25, < or = 65 years), and older patients, O (n = 21). Changes in AP and HR after the first minute of tilting, during the stable orthostatic phase and during syncope were compared. Except for systolic pressure, baseline hemodynamic parameters were similar in Y and O. No difference appeared in the mean time elapsed before syncope (19+/-9 vs 22+/-2 min). Asymptomatic hypotension was observed, only in O, 1 minute after tilting, followed by a progressive fall in the mean AP before syncope (0+/-0.9 vs -1+/-0.7 mmHg/min) without HR increase (0.7+/-1 vs 0+/-0.6 beats/min). This pressure slope was strongly related to age (r = 0.54, P < 0.001). Hemodynamic recording during HUT identifies a dysautonomic pattern in elderly patients with NCS and the abnormal AP/HR responses to orthostasis may be a feature specific to this population. Although the central mechanism of NCS is common to all ages, the age-related characteristics of the trigger event may indicate the need for specific management at different ages. PMID- 10456629 TI - Dual chamber pacing with a single-lead DDD pacing system. AB - The successful application of single-lead VDD pacing during the last few years has generated the idea of single-lead DDD pacing. Preliminary data from several single-lead VDD studies attempting to pace the atrium by a floating atrial dipole are unsatisfactory, causing an unacceptably high current drain of the device. We studied the feasibility as well as the short- and long-term stability of atrioventricular sequential pacing, using a new single-pass, tined DDD lead. In eight consecutive patients (age 73+/-16 years) with symptomatic higher degree AV block and intact sinus node function, this new single-pass DDD lead was implanted in combination with a DDDR pacemaker. Correct VDD and DDD function was studied at implantation; at discharge; and at 1, 3, and 6 months of follow-up. At implant, the atrial stimulation threshold was 0.6+/-0.1 V/0.5 ms. During follow-up, the atrial pacing thresholds in different every day positions averaged 2.1+/-0.5 V at discharge, 2.9+/-0.5 V at 1 month, 3.8+/-0.4 V at 3 months, and 3.4+/-0.4 V at 6 months (pulse width always 0.5 ms). The measured P wave amplitude at implantation was 4.5+/-2.2 mV; during follow-up the telemetered atrial sensitivity thresholds averaged 2.1+/-0.3 mV. Phrenic nerve stimulation at high output pacing (5.0 V/0.5 ms) was observed in three (38%) patients at discharge and in one (13%) patient during follow-up; an intermittent unmeasurable atrial lead impedance at 3 and 6 months follow-up was documented in one (13%) patient. This study confirms the possibility of short- and long-term DDD pacing using a single-pass DDD lead. Since atrial stimulation thresholds are still relatively high compared to conventional dual-lead DDD pacing, further improvements of the atrial electrodes are desirable, enabling lower pacing thresholds and optimizing energy requirements as well as minimizing the potential disadvantage of phrenic nerve stimulation. PMID- 10456630 TI - Neurohormonal profile before and after beta-blockade in patients with neurocardiogenic syncope. AB - Our objective was to evaluate the effects of beta-blockers on the neurohormonal profile, particularly vasopressin (VP) release, in vasovagal syncope and to gain further insight on the pathophysiology of this syndrome. Patients (< or =75 years) with no cardiovascular, neurological disorders, or contraindications to the use of isoproterenol or beta-blockers and being explored for unexplained syncope were included. An 80 degrees HUT was performed under identical conditions. After a 25-min period of passive tilt, isoproterenol was infused at a rate of 1-5 microg/mn if required. Two groups matched for age and sex were considered: a HUT-positive and a HUT-negative group. The HUT-positive group was then given beta-blockers, subsequently reassessed, and divided into two subgroups: alpha beta-blocker nonresponder group and a beta-blocker responder group. Blood samples for assays of norepinephrine (NE), epinephrine (E), and VP were taken at baseline and the end of the procedure. In all, 44 subjects entered the study, 22 in each group. The HUT-positive group exhibited an obvious lesser increase in plasma NE and a clear-cut rise in plasma E and VP compared to the HUT negative group (P < 0.05). Even though no patient in the HUT-positive group reported recurrent symptoms under treatment, the second HUT could distinguish two subgroups: a beta-blocker nonresponder group (n = 12) whose HUT remained positive and a beta-blocker responder group (n = 10) whose HUT was normalized. The time course of plasma E and VP during the second HUT was similar to that for the HUT positive and HUT-negative groups. In conclusion, the efficacy of beta-blockers is associated not only with a reduction of the sympathoadrenal stimulation seen in vasovagal syncope but also with a lower release of VP suggesting that low pressure baroreceptors might be involved in VP release. PMID- 10456631 TI - Time dependence of anodal and cathodal refractory periods in cardiac tissue. AB - Mehra et al. (PACE 1980; 3:526) observed that immediately after implantation of a pacing electrode in a dog heart, the anodal refractory period (RP) is shorter than the cathodal RP, but after several weeks the anodal RP becomes longer than the cathodal RP. We examine this experiment using numerical simulations based on the bidomain model of cardiac tissue and a Beeler-Reuter membrane. Our hypothesis is that accumulation of inexcitable tissue around the electrode following implantation causes the effective size of the electrode to increase and that this increase is the mechanism underlying the change in RP. We calculate that the anodal RP is shorter than the cathodal RP for both large and small electrodes. However, for large electrodes the threshold for anode "break" stimulation is greater than 8 mA. Mehra et al. defined RP experimentally as the interval at which the threshold stimulus strength becomes greater than 8 mA. If we restrict the stimulus current in our calculations to less than 8 mA, we exclude anode break stimulation from our calculation of the RP. In that case, our results are consistent with Mehra et al. and suggest that their observation resulted from their definition of RP. PMID- 10456632 TI - The potential for inappropriate ventricular tachycardia confirmation using the Intracardiac Electrogram (EGM) Width Criterion. AB - The "Intracardiac Electrogram (EGM) Width Criterion," the first digital signal processing feature used in an implantable cardioverter defibrillator (ICD), is a detection enhancement algorithm that intends to improve ventricular tachycardia (VT) detection specificity by rejecting inappropriately detected supraventricular tachyarrhythmias. The algorithm may be activated after setting the optimal EGM source, slew, and width thresholds based on EGM width testing during sinus rhythm. This study evaluates the accuracy of the EGM width measurements during exercise testing. Twenty-one patients with Medtronic Micro Jewel II Model 7223 ICDs underwent treadmill exercise testing. EGM width testing was repeatedly performed during exercise and recovery to detect potential inappropriate measurements. In seven (33%) patients the EGM Width Criterion inappropriately confirmed VT detection. Eleven patients had inappropriately wide EGM width measurements, but did not satisfy the EGM Width Criterion. The causes of wide EGM width measurements were an actual increase in EGM width and/or inappropriate detection of the baseline irregularities as EGM onset or offset points. Based on our observations, we recommend to test the EGM Width Criterion during exercise testing for optimal ICD programming. PMID- 10456633 TI - Aortic input impedance and ventriculoarterial coupling following cardioversion/defibrillation. AB - Defibrillation shocks are commonly used after cardiac surgery or during defibrillator implantation. The hemodynamic consequences of countershocks on circulatory dynamics are not completely understood, and there is a lack of information concerning the effects on ventriculoarterial interaction. The study presented here was performed to assess the influence of defibrillation shocks on arterial hemodynamics and ventriculoarterial coupling. Eight mongrel dogs (weight 15-18 kg) were anesthetized and median thoracotomy was performed. Pressure in the ascending aorta and the left ventricle and flow in the ascending aorta were continuously monitored. After induction of atrial or ventricular fibrillation, termination was achieved by epicardial low energy shocks (atrium, 3J; ventricle, 51). In an additional attempt ventricular fibrillation was terminated by a high energy shock (34J). Aortic input impedance was calculated by fast-Fourier transformation of aortic flow and pressure signals, while ventriculoarterial coupling was expressed by the ratio of aortic and ventricular end systolic elastance (Ea/Ees). Defibrillation by low energy shocks of atrial or ventricular fibrillation did not result in changes of the aortic impedance spectrum, and ventriculoarterial coupling remained unaltered compared to control conditions. High energy defibrillation, however, resulted in a marked rise in total peripheral resistance (P < 0.03). The ratio of Ea/Ees increased significantly (P < 0.005). These effects were reversible within 15 minutes. Low energy defibrillation does not alter arterial hemodynamics and ventriculoarterial coupling in this experimental setup. High energy defibrillation, however, results in a temporary increase of ventricular load. This finding may be of particular interest in patients with poor left ventricular function. PMID- 10456634 TI - Relationship between atrioventricular delay and oxygen consumption in patients with sick sinus syndrome: relevance to rate responsive pacing. AB - To develop a dromotropic-controlled rate adaptive algorithm for patients with sick sinus syndrome (SSS) and intact AV conduction, 14 pace-maker patients with SSS underwent cardiopulmonary exercise testing (CPX). During exercise, the pace maker was programmed in an AAT mode without rate adaptation, whereby 3 patients developed supraventricular arrhythmia and 11 patients kept sinus rhythm. Chronotropic incompetence (CI) at heart rate (HR) < 95 beats/min at the anaerobic threshold (AT) was found in five patients. In patients with chronotropic competence (CC), the HR increase was significantly greater than in CI patients (rest: 73.2 +/- 12.6 vs. 64.2 +/- 4.0 beats/min;AT:101.2 +/- 6.2 vs. 82.0 +/- 5.1 beats/min;peak: 135.2 +/- 10.7 vs. 103.2 +/- 10.9 beats/min). There was no significant difference in the AVD between CC and CI patients (rest: 167.7 +/- 38.6 vs. 170.8 +/- 22.5 ms, AT: 156.2 +/- 30.7 vs. 163.6 +/- 21.6 ms, peak: 144.7 +/- 29.0 vs. 152.4 +/- 15.0 ms). The correlation coefficient between HR increase and VO2 was +1.0 and between AVD decrease and VO2 - 1.0 in both groups. An increase in pacing rate from 75 beats/min to 120 beats/min without exercise (overpacing) led to a prolongation of the AV interval of about 30.6 +/- 14.2 ms. Based on this closed loop control with negative feedback, a dromotropic rate adaptive algorithm for patients with SSS and intact AV conduction could be developed. PMID- 10456635 TI - Vasoactive peptide release in relation to hemodynamic and metabolic changes during rapid ventricular pacing. AB - Plasma atrial natriuretic peptide (ANP) concentration increases during ventricular arrhythmias and rapid ventricular pacing but less is known about plasma brain natriuretic peptide (BNP) and endothelin (ET-1). In the present study concentrations of ANP, the amino terminal part of the proANP (NT-proANP), BNP, and ET-1 were measured in the coronary sinus and femoral artery before and at the end of rapid ventricular pacing in 15 patients with coronary arterial disease. The changes were compared with the changes in mean arterial blood pressure, pulmonary capillary wedge pressure (PCWP), transcardiac differences in pH, pCO2, lactate, and norepinephrine. There was an increase in PCWP and a transient decrease in blood pressure after initiation of pacing. Pacing caused a decrease in ST-segment, transcardiac difference of norepinephrine, lactate extraction, pCO2 difference, and an increase in pH difference. Concentration of ANP in the coronary sinus and femoral artery and its transcardiac difference increased during pacing (P < 0.001), whereas changes in NT-proANP were small and BNP and ET-1 levels remained unchanged. The change in transcardiac ANP difference correlated with the change in lactate (r = 0.53, P < 0.05) but not that of norepinephrine, PCWP, or blood pressure. The results show that the plasma concentration of ANP increases more than that of NT-proANP during rapid ventricular pacing. Ischemia-induced release of ANP and its diminished elimination may contribute to the increased plasma ANP level. PMID- 10456636 TI - Low temperature and low energy radiofrequency modification of atrioventricular nodal slow pathways in pediatric patients. AB - OBJECTIVES: To report our experience using low temperature and energy in the modification of the slow pathway in pediatric patients with atrioventricular nodal reentrant tachycardia. BACKGROUND: A concern in performing a slow pathway modification is the possible damage of the normal AV conduction system. Lesion size has been shown to have a linear relationship with temperature. Previous reports have used energy of 25-50 W that generate temperatures of 60 degrees C 70 degrees C for successful procedures. METHODS: Report of results of attempted AV nodal slow pathway modification in 17 consecutive pediatric patients < 15 years of age at The Children's Hospital of Philadelphia from April 1995 to November 1997 using low temperature and energy. RESULTS: There were 18 successful slow pathway modifications with 1 recurrence in 17 patients. The maximum energy used during successful lesions was 32.7 +/- 13.8 W (range 15-50 W) with a mean energy of 26.4 +/- 13.3 W (range 12-48 W). The peak temperature during these lesions was 55.1 degrees C +/- 4.1 degrees C (range 48 degrees C-64 degrees C) with a mean temperature of 47.9 degrees C +/- 2.7 degrees C (range 44 C-540C). The mean number of radiofrequency lesions required for a successful modification was 5.8 +/- 6.7 (median 4.0, range 1-26). Patients have been followed for 2.08 +/ 0.79 years. CONCLUSIONS: Slow pathway modification can be performed successfully with a low incidence of recurrence in the pediatric patient using low energy and temperature. It is possible that this may lead to smaller lesions. PMID- 10456637 TI - Clinical surveillance of a tined, bipolar, J-shaped, steroid-eluting, silicone insulated atrial pacing lead. AB - Since 1990, 558 Medtronic 5524 bipolar, silicone-insulated, J-shaped, tined, steroid-eluting atrial leads have been implanted at the Mayo Clinic (Rochester, MN, USA) and the Midelfort Clinic (Eau Claire, WI, USA). Implantation data were favorable, with pacing thresholds at implantation (median threshold, 0.6 V) better than most published data on other atrial leads. The rate of acute lead related complications (dislodgment and diaphragmatic pacing) necessitating reoperation or electrical abandonment of the atrial lead was 0.9%. This rate is lower than that in most published series of atrial leads. Over a median follow-up time of 17.5 months (up to 69 months), there were no chronic lead-related complications and no definite or suspected failures of lead material. This rate is much lower than that with other atrial leads studied previously. We conclude that the Medtronic 5524 atrial lead combines the reliability of silicone insulation with a lack of chronic complications and high thresholds due to its steroid elution and with stability in the atrium due to its J shape despite a passive fixation mechanism. There is no evidence of lead material failure during up to 6 years of follow-up. PMID- 10456638 TI - Impact of long-term ECG recording on the detection of paroxysmal atrial fibrillation in patients after an acute ischemic stroke. AB - An ECG recording time of 24 hours has a low yield to detect atrial arrhythmias in patients after an acute ischemic stroke. The present study investigated whether a recording time of 72 instead of 24 hours detects paroxysmal atrial fibrillation in more patients. The study prospectively included 82 consecutive patients 2-3 weeks after an acute ischemic stroke. All patients had sinus rhythm in the resting ECGs and no history of atrial fibrillation or flutter. The frequency of atrial fibrillation was assessed after 24, 48, and 72 hours of ambulatory ECG monitoring. An ECG monitoring time of 72 hours documented paroxysmal atrial fibrillation in five (6%) patients. The episode of paroxysmal atrial fibrillation occurred in only one patient within 24 hours. The other patients had their first episode of atrial fibrillation between 24 and 48 hours (n = 2) and between 48 and 72 hours (n = 2). These five patients were older (age = 70 +/- 5 years), whereas the mean age of the remaining patients was 59 +/- 13 years. All five patients had cardiovascular disease in comparison to 36 of 77 patients and reported palpitations in comparison to 6 of 77 of the remaining patients. In conclusion, ambulatory ECG monitoring over 72 hours detected after the first recording day four of five patients in whom paroxysmal atrial fibrillation could be documented for the first time. The 72-hour recording time improved, compared to the 24-hour period, the detection of paroxysmal atrial fibrillation in patients after an ischemic stroke. It seems to be more efficient to perform prolonged ECG recording mainly in older patients with a cardiovascular disease and/or a history of palpitations. PMID- 10456639 TI - Blind axillar venous access. AB - The axillary vein has been suggested as an alternate site for venous access to avoid the "subclavian crush phenomenon. " Many techniques have been used to access this structure. They include complicated anatomical landmarks, contrast venography, Doppler, and ultrasound. A simple technique using the basic anatomical landmark of the deltopectoral groove and a blind venous stick has been used successfully in 165 of 168 consecutive pacemaker and ICD procedures; there were only three failures. These required an alternate approach. With a thorough knowledge of the regional anatomy, the axillary vein can be safely used as a primary site of venous access. PMID- 10456640 TI - Narrow QRS tachycardia with changing R-P relationship. PMID- 10456641 TI - Arrhythmogenic right ventricular dysplasia in an American Indian woman. AB - Arrhythmogenic right ventricular (RV) dysplasia consists of a dilatation of the right ventricle with a reduction of RV ejection fraction with fibrofatty replacement of the RV myocardium in the face of a well-preserved left ventricular systolic function. Arrhythmogenic RV dysplasia, which is a cause of sudden unexpected death, has been reported from many geographic areas, including the United States, Europe, and the Far East. This case report presents the first case of arrhythmogenic RV dysplasia in an American Indian (Native American) patient. PMID- 10456642 TI - Treatment of advanced atrioventricular block with beta-adrenergic blockade therapy. AB - A 26-year old woman, who experienced syncope associated with advanced AV block, was referred for further evaluation. Electrophysiological study showed normal SA and AV node physiology. Syncope associated with advanced AV block on the electrocardiogram was induced by head-up tilt test. Oral propranolol therapy completely prevented the AV block and syncope induced by head-up tilt. The physician should be aware of advanced AV block associated with neurocardiogenic syncope, especially before permanent - pacemaker therapy is considered. The head up tilt test is a useful diagnostic tool for an etiology of paroxysmal AV block. PMID- 10456643 TI - Percutaneous extraction of an abandoned coiled pacing lead. AB - Abandoning redundant pacing leads has been an accepted practice in most cases. Late migration of a lead is a rare, unpredictable, and potentially lethal complication of this approach and requires immediate removal. We report the case of an atrial pacing lead that had migrated and was found coiled in the right atrium. It was removed by a snare catheter via a femoral approach. PMID- 10456644 TI - Absence of the superior vena cava: difficulties for pacemaker implantation. AB - Endocardial pacemaker electrode implantation can be difficult in patients with anomalous superior vena cava (SVC). Venography and CAT scan showed that the patient lacked SVC venous drainage and that systemic veins drained into the inferior vena cava through the azygos vein. A temporary stimulation electrode was placed by puncture of the femoral vein, permanent stimulation by venotomy of the epigastric vein, with the electrode inserted through the external iliac vein. PMID- 10456645 TI - Pacemaker interference with screening mammography. AB - In female pacemaker recipients undergoing screening mammography, the impact of a pulse generator placed in the pectoral region has yet to be reviewed. We evaluated mammograms from 74 female pacemaker patients aged 40 years and older. The pulse generator obscured a portion of the mammogram in 7 patients (12%). During pacemaker implantation in women, the potential for the pulse generator to interfere with screening mammography should be considered. Baseline mammography should be obtained or reviewed; in high risk patients a nonconventional location for the pulse generator may be appropriate. PMID- 10456646 TI - Optic neuropathy following amiodarone therapy. AB - Ocular changes during treatment with amiodarone are almost universal but are rarely serious. In this article we describe three patients from a single electrophysiology practice in whom optic neuropathy developed during treatment with amiodarone. All three patients were more than 65 years of age. The doses of amiodarone ranged from 100 to 400 mg/day, and the time intervals between the initiation of the amiodarone therapy and the appearance of first symptoms of optic neuropathy were 5 to 19 months. Two patients had bilateral involvement, and one had only unilateral involvement. Whether this result was due solely to amiodarone therapy, to the underlying poor health of these patients, or to the combination of these two factors is uncertain. These findings prompt us to recommend that all patients who receive amiodarone undergo complete ophthalmologic examinations, including careful evaluation of the ocular fundus regularly during such therapy. No randomized study had been undertaken to determine the true incidence of complications associated with this medication. PMID- 10456647 TI - Atrial epicardial pacing with long stimulus to P wave interval in a patient with arrhythmogenic right ventricular dysplasia complicated by right atrial thrombosis. AB - Atrial epicardial pacing with a long stimulus to P wave interval in a patient with arrhythmogenic right ventricular dysplasia complicated by right atrial thrombosis is discussed. Arrhythmogenic right ventricular dysplasia (ARVD) is associated with a high incidence of malignant ventricular arrhythmias. Most patients with ARVD need antiarrhythmic drugs, catheter ablation, or an implantable cardioverter defibrillator. We report a patient with ARVD in whom effective treatment with sotalol caused severe, symptomatic sinus bradycardia requiring permanent pacing. Due to leftward displacement of the right ventricle and the presence of two thrombi in the right atrium, an epicardial atrial lead and AAI pacemaker were implanted. A long stimulus to P wave interval caused by severe dilatation of the right atrium was recorded. During a 6 months of follow up on sotalol treatment there were neither ventricular tachycardia (VT) attacks nor pacing problems. PMID- 10456648 TI - Dynamic left ventricular arterial pressure gradient and sick sinus syndrome with heterozygous Fabry's disease improved following implantation of a dual chamber pacemaker. AB - A 63-year-old woman with heterozygous Fabry's disease, sick sinus syndrome, sinus pauses, congestive heart failure, syncope, and a dynamic pressure gradient between the left ventricle and systemic arteries underwent implantation of a dual chamber (DDD) pacemaker. Following pacemaker implantation, the pressure gradient has been reduced and congestive heart failure controlled for the past 3 years. PMID- 10456649 TI - Normal sinus rhythm in a patient with corrected transposition of great arteries after 30 years of complete heart block. AB - A case of a patient with corrected transposition of great arteries with complete atrioventricular block for 29 years who was in sinus rhythm. PMID- 10456650 TI - Perforation of the right ventricular free wall by an active fixation transvenous cardioverter defibrillator lead. AB - A 58-year-old patient with dilated cardiomyopathy underwent implantable cardioverter defibrillator (ICD) implantation. The postoperative course was complicated by perforation of the right ventricular free wall by the active fixation transvenous ICD lead. The type of ICD lead and the type of organic heart disease are apparently important risk factors for perforation. PMID- 10456652 TI - The Stimarec Bulletin. PMID- 10456651 TI - Implantable defibrillator endocarditis. PMID- 10456653 TI - The Stimarec Bulletin. PMID- 10456654 TI - Mechanisms of polyclonal hypogammaglobulinaemia in multiple myeloma (MM). PMID- 10456655 TI - Cancer gene and immunotherapy: recent developments. AB - Gene and immunotherapeutic approaches to treat human malignant tumors are reviewed. Special attention is given to the different strategies of cancer gene therapy and to recent aspects of cytokine-supported tumor immunotherapy or tumor specific vaccination. The limitations of these therapy approaches are critically discussed especially with respect to immune escape mechanisms. PMID- 10456657 TI - Locally advanced breast cancer in Saudi Arabia: high frequency of stage III in a young population. AB - In the Kingdom of Saudi Arabia (KSA), breast cancer constitutes 18% of all cancers in Saudi women. Whilst locally advanced breast cancer disease is unusual in Western countries, it constitutes more than 40% of all non-metastatic breast cancer in KSA. The relative frequency of locally advanced disease among our breast cancer population and the lack of a uniform consensus in the literature about its optimal management have prompted this retrospective analysis of the medical records of patients with Stage III breast cancer patients seen at King Faisal Specialist Hospital and Research Center between 1981 and 1991. In all, 315 patients were identified. Their median age +/- SD was 46 +/- 11.6 years which is distinctly different from the 60-65 years median age in industrial Western nations. Most patients were younger than 50 years (64%) and premenopausal (62%). Patients were approximately equally divided between Stage IIIA and Stage III B. Patients received multimodality treatment, including surgery, adjuvant chemotherapy, tamoxifen, and adjuvant radiotherapy. Sixty-one patients were excluded from survival analysis as they were considered lost to follow-up. Of the remaining 254 patients, 73 (29%) were alive and disease free, and 18 patients (7%) were alive but with evidence of the disease. The remaining 163 (64%) had died from breast cancer or its related complications. Their median overall survival (OS) was 54 months, (95% CI, 27 to 121 months) and the median progression-free survival (PFS) was 28.8 months (95% CI, 14.2 to 113 months). Cox proportional hazard model identified Stage III B and the number of positive axillary lymph nodes as poor predictors of OS and PFS. Radiotherapy was the only adjuvant modality that affected survival favourably. The prognosis of patients with Stage III disease remains poor despite the use of a multimodality approach. The overall young age of our patients may have contributed to the poor outcome. Moreover, the adverse effect of Stage III B disease (as compared with Stage III A) and axillary nodal status was evident. Whilst the favourable effect of radiotherapy on survival was demonstrated, the lack of independent efficacy of other modalities (adjuvant chemotherapy and tamoxifen) or the apparent deleterious effect of neoadjuvant chemotherapy should be addressed with discretion in such retrospective analysis. Optimal management of patients with locally advanced breast cancer disease should be appraised in well designed, prospective, randomised studies. PMID- 10456656 TI - Interleukin-10: a cytokine used by tumors to escape immunosurveillance. AB - The mechanisms whereby malignant cells can elude the recognition of the immune system, by what is termed 'immunological escape', have attracted major attention of tumor immunologists during the past decade. In this review, the role of the immunosuppressive cytokine interleukin-10 (IL-10) will be discussed as a strategy used by tumors to avoid recognition by cytotoxic T lymphocytes (CTL). PMID- 10456658 TI - Phase II trial of high-dose dexamethasone for untreated patients with primary systemic amyloidosis. AB - Primary systemic amyloidosis is the disorder that results from the deposition of insoluble immunoglobulin light chain fragments. Patients seen within 30 days of diagnosis have a median survival of 13 months. Patients treated with melphalan and prednisone have a median survival of 17 months. There is a need for new therapies. A prospective study was undertaken of high-dose dexamethasone in the treatment of 25 patients with previously untreated primary systemic amyloidosis. Treatment was similar to that given to patients with multiple myeloma. In this cohort, three patients showed objective regression with organ-specific improvement of the disease. The median survival of the entire group was 13.8 months. High-dose dexamethasone is of occasional benefit in patients with amyloidosis but does not appear to be superior to melphalan and prednisone chemotherapy. PMID- 10456659 TI - Transforming growth factor-beta and multidrug resistance in chronic lymphocytic leukemia. AB - Patients with chronic lymphocytic leukemia (CLL) frequently respond to initial treatment, but then become resistant to chemotherapy. Studies have shown one important cause of chemotherapeutic resistance to be multidrug resistance (MDR). To investigate the potential role of MDR and transforming growth factor-beta (TFG beta), a potent growth inhibitor of B lymphocytes, in the development of chemotherapeutic resistance in CLL, we evaluated 22 CLL patients for loss or mutation of TGF-beta receptors (TbetaR), plasma TGF-beta1 levels, and expression of MDR1 mRNA. Receptor crosslinking and immunoprecipitation experiments did not demonstrate loss of TbetaRs in any patients studied. No relationship between plasma TGF-beta1 levels and expression of MDR1 mRNA was seen. Correlation of plasma TGF-beta1 levels to disease stage revealed a consistent decline in plasma TGF-beta1 levels with advancing disease stage (P = 0.031). PMID- 10456661 TI - Paraneoplastic syndromes in 68 cases of resectable non-small cell lung carcinoma: can they help in early detection? AB - The aim of this study was to assess the importance of paraneoplastic syndromes as an early sign of non-small cell lung cancer (NSCLC). A procedure for searching paraneoplastic syndromes, based on 40 years of reports in the literature, was established and the prevalence of paraneoplastic syndromes estimated in 68 patients with resectable NSCLC. Stages I and II were considered eligible for surgery straight away. Patients in Stage IIIA underwent surgery if partially or completely responsive to three courses of neo-adjuvant chemotherapy. Paraneoplastic syndromes were assessed and confirmed in nine patients (13%). Motor-sensory neuropathy, arthritis and arthralgias to the knees, periarthritis to the shoulder, hypertrophic osteopathy, clubbing, pruritus were observed. Only three patients with painful osteoarthropathies were diagnosed with NSCLC by tracing their paraneoplastic syndrome, whereas most of them (36/68) were diagnosed incidentally through a chest radiograph taken for tumour-unrelated symptoms. A careful research of paraneoplastic syndromes in high risk patients may guide the doctor to a resectable NSCLC diagnosis. Recent onset arthritis and arthralgias, which cannot be explained otherwise, should be considered to be early clues of lung cancer. PMID- 10456660 TI - Assessment of all-trans retinoic acid (ATRA) efficacy as a single agent in primary lymphoid neoplasia. AB - All-trans retinoic acid (ATRA) is currently widely used in the therapy of acute promyelocytic leukemia and is being tested in vitro and in vivo on several other malignancies. Previously ATRA has been shown to inhibit the growth in vitro, of established human myeloma cell lines as well as cultured primary myeloma cells from patients. ATRA acts by down-regulating IL-6-receptor-alpha or gp130 on the surface of the myeloma cells. However, despite its in vitro effects on myeloma cells, ATRA therapy on advanced stage multiple myeloma (MM) patients has so far largely been ineffective. In current studies, we have assessed the efficacy of ATRA therapy against primary murine plasma cell tumors, which are an animal model for human MM. These tumors are induced at about 50% incidence in pristane-primed BALB/c mice by injection of v-raf/v-myc- containing retroviruses and are IL-6 dependent. Using this animal model, we assessed the effect of ATRA as a therapeutic agent against primary tumors at two early time points in disease development. ATRA was administered in liposomal vesicles (ATRAGEN), since liposomal-ATRA has been shown to circumvent clearance mechanisms by hepatic microsomes, which normally occur with free ATRA. In addition, ATRAGEN was previously shown to be less toxic in mice than free ATRA. ATRAGEN was administered beginning on day 25 or day 45 after virus injection and continued twice weekly for 8-11 weeks. ATRAGEN administration begun at either time point did not alter the incidence or the latency of plasma cell tumors compared with control animals. These results suggest that ATRA may not be an effective sole therapy against early MM. PMID- 10456662 TI - Concurrent occurrence of three neoplasms including non-Hodgkin's lymphoma, renal cell carcinoma and leiomyoma in the same kidney. AB - A 53-year-old man with triple renal neoplasms in his left kidney presented. He was initially diagnosed intermediate grade non-Hodgkin's lymphoma (NHL) which involved gastrointestinal tract, left kidney, liver and pancreas. He underwent left nefrectomy because of a persistent renal mass after the completion of chemotherapy. The large renal mass revealed a renal cell carcinoma (RCC). Additionally, multiple small nodules of non-Hodgkin's lymphoma and a solitary leiomyoma were observed. PMID- 10456663 TI - Hepatosplenic candidiasis after neutropenic phase of acute leukaemia. AB - Hepatosplenic candidiasis following granulocytopenic periods is a relatively recently recognised problem in immunocompromised patients, particularly in those with acute leukaemia. We present three patients in whom diagnosis of hepatosplenic candidiasis was suspected on the basis of ultrasonographic (US), computed tomographic (CT) findings and confirmed by laparoscopy and biopsy of liver lesions. All three patients were successfully treated briefly with amphotericin B, followed by a longer period of fluconazole. In one patient laparotomy and surgical evacuation of abscesses was performed. This condition could be more often recognised by careful follow-up of liver function test, C reactive protein level, ultrasonography, CT and MRI after recovery from chemotherapy-induced neutropenia. PMID- 10456664 TI - Histocompatibility testing guidelines for hematopoietic stem cell transplantation using volunteer donors: report from The World Marrow Donor Association. Quality Assurance and Donor Registries Working Groups of the World Marrow Donor Association. AB - The World Marrow Donor Association has formulated guidelines for establishing the extent and quality of histocompatibility testing for unrelated donor registries, umbilical cord blood banks, and transplant centers involved in international exchange of hematopoietic stem cells for allogeneic transplantation. The ability to identify unrelated stem cell donors in one country for patients in another country requires cooperation and standardization in many areas. The adoption of guidelines for histocompatibility testing, such as those summarized in this report, will facilitate these opportunities and rapidly provide accurately typed donors for patients in need. PMID- 10456665 TI - A proposal regarding the mechanism which underlies lineage choice during hematopoietic differentiation. AB - We present a simple Boolean Network model of a genetic regulatory network which can account for the differentiation of hemopoietic cells. In this model while decision choice is stochastic, the probability of differentiating along a specific pathway is determined by the gene expression state of the cell when the signal to differentiate appears. This probability can be altered by the acute and chronic needs of the individual. Mutations disrupt normal maturation. The paper also presents a larger Boolean Network model which can be used to study normal and leukemic stem cell attractor states. PMID- 10456666 TI - Salvage therapy of refractory and relapsed acute leukemia with high dose mitoxantrone and high dose cytarabine. AB - We have assessed the outcome of 66 refractory and relapsed acute leukemia patients treated with high dose mitoxantrone and cytarabine. Therapy consisted of a total dose of 40-60 mg/m2 mitoxantrone and 3 g/m2 of cytarabine daily on 5 consecutive days. A total of 28 patients were treated for primary resistant and 38 patients for early or late relapsed leukemia. A total of 35 patients achieved CR. Four patients died during the induction course. Toxicity was acceptable and comparable to other salvage regimens. The median disease-free and overall survivals were 4 and 6 months, respectively. Although this regimen is effective in achieving remission in refractory leukemia, its duration is short. PMID- 10456667 TI - The importance of cell size and surface marker analysis in childhood acute myeloblastic leukemia. AB - In order to evaluate the prognostic significance of cell size and surface marker expression, we evaluated 33 children with newly diagnosed acute myeloblastic leukemia by flow cytometry. We determined: the percentage of small, middle and large cells; large to small cell ratios (LS); large plus middle to small cell ratios (LMS); the percentage of surface markers expressed by each group of cell; the ratios of surface marker percentages expressed by the large blasts to that expressed by small blasts (LS for surface markers); and large plus middle blasts to that by small blasts (LMS for surface markers). For 'early prognosis', patients who could and could not achieve remission (n = 23 and 10) and for late prognosis, the patients who deceased or relapsed within the first 12 months of the treatment (n = 24) and who survived for more than 12 months (n = 9) were compared, in two classifications. CD3 percentages of the small cells of alive patients were significantly higher than that of dead or relapsed patients. LMS for CD3 and CD20 and LS for CD20 were higher in dead relapsed patients than that of alive patients. The total percentage of CD14 was significantly higher in dead relapsed patients than it was in the alive patients and CD3 was significantly higher in the group of patients who achieved remission than that of the patients who could not achieve remission. It was striking that, expression of CD3, CD7, CD22, CD33, CD14, CD15, CD34 increased or decreased as to cell size, whatever the prognosis. CD10, CD20 and CD13 were expressed on the large cells of the patients who could not achieve remission or died relapsed. We showed that, the blast cell size, individually does not have any prognostic significance in childhood AML and the prognostic significance of surface markers not only depends on their presence or absence but also on their relative configuration of expression by the blasts with different size. PMID- 10456668 TI - Delphi-panel analysis of appropriateness of high-dose therapy and bone marrow transplants in adults with acute myelogenous leukemia in 1st remission. AB - BACKGROUND: Despite considerable data, there is still controversy over which adults with acute myelogenous leukemia (AML) in 1st remission should receive high dose therapy and a bone marrow transplant rather than conventional-dose chemotherapy. Analyses of data from randomized trials are complex, conclusions sometimes contradictory and results not sufficiently detailed to allow subject level decisions. OBJECTIVE: To determine appropriate use of high-dose therapy and bone marrow transplants in persons with AML in 1st remission with specific features. Develop a treatment algorithm. PANELISTS: Nine leukemia experts from diverse geographic sites and practice settings. EVIDENCE: Boolean MEDLINE searches of acute myelogenous leukemia and chemotherapy and/or transplants. CONSENSUS PROCESS: We used a modified Delphi-panel group judgment process. Age, WBC, cytogenetics and FAB-type were permuted to define 72 clinical settings. Each panelist rated appropriateness of high-dose therapy and a transplant versus conventional-dose chemotherapy on a nine-point ordinal scale (1, most inappropriate, 9, most appropriate) considering 3 types of donors: (1) HLA identical siblings; (2) alternative donors (HLA-matched related or unrelated people other than an HLA-identical sibling); and (3) autotransplants. An appropriateness index was developed based on median rating and amount of disagreement. The relationship of appropriateness indices to the permuted clinical variables was considered by analysis of variance and recursive partitioning. Preference between donor types was analyzed by comparing mean appropriateness indices of comparable settings and a treatment algorithm developed. CONCLUSIONS: In people with an HLA-identical sibling, this type of transplant was rated appropriate in those with unfavorable cytogenetics and uncertain in all other settings. In people without an HLA-identical sibling, an alternative donor transplant was rated appropriate in those < 30 years with unfavorable cytogenetics, uncertain in those > 30 years and unfavorable cytogenetics and inappropriate in all other settings. Autotransplants were rated appropriate in people with unfavorable cytogenetics and uncertain in all other settings. An HLA-identical sibling donor, when available, was always preferred to an alternative donor transplant or autotransplant. In people without an HLA identical sibling, an autotransplant was almost always favored over an alternative donor transplant with the magnitude of preference inversely correlated with transplant appropriateness. PMID- 10456669 TI - Secondary acute myeloid leukemia 4 years after the diagnosis of hairy cell leukemia: case report and review of the literature. AB - A 49-year-old man diagnosed with hairy cell leukemia (HCL) achieved a complete remission lasting 4 years after treatment with cladrabine and subsequently developed acute myeloid leukemia. Although a wide variety of second malignancies have been noted in HCL with an incidence of 8.7%, acute myeloid leukemia (AML) has been reported only once previously in a splenectomized patient who had been treated with alpha interferon. PMID- 10456670 TI - Megakaryocyte induced fibroblast proliferation is enhanced by costimulation with IL-6/IL-3 and dependent on secretory and adhesion events. AB - Experimental data are in keeping with the finding that megakaryocytes isolated from normal human bone marrow may promote fibroblast growth. This effect can be significantly enhanced by interleukin (IL)-3. In this context it has been demonstrated that IL-3 induces the release of platelet-derived growth factor (PDGF) and transforming growth factor beta1 (TGFbeta1) from megakaryocytes, factors known to enhance fibroblast proliferation. The present in vitro study was performed to elucidate the action of several other cytokines which are able to influence the different steps of megakaryocyte maturation and function like stem cell factor (SCF), IL-6, and leukemia inhibitory factor (LIF) as well. Following an appropriate experimental design we were able to show that none of the mentioned cytokines enhanced megakaryocyte dependent fibroblast proliferation in the coculture assays. On the other hand, IL-6 in combination with IL-3 surpassed the IL-3 dependent action significantly. However, the combined IL-3/IL-6 effect was not explainable by an increased PDGF/TGF-beta secretion of the megakaryocytes. In transwell experiments the inhibition of cell-to-cell contact via tissue culture inserts generated a conspicuous impairment of fibroblast growth in the IL-3/IL-6 treated cocultures. This reversal surpassed even the effect on the untreated and IL-3 stimulated cocultures. Hence, a direct contact of both cell types probably inducing adhesion phenomenons and warranting a certain threshold of local PDGF/TGF-beta concentration is a prerequisite for the proliferative effect on fibroblasts in the costimulation experiments. These results are of special interest regarding the evolution of myelofibrosis in chronic myeloproliferative disorders (CMPDs) because (1) various progenitor cells including the megakaryocytic lineage are hypersensitive towards IL-3 and (2) an abnormal secretion of IL-6 is described for megakaryocytes in these disorders. PMID- 10456671 TI - A novel biphenotypic B-cell precursor leukemia cell line (NALM-29) carrying t(9;22)(q34;q11) established from a patient with acute leukemia. AB - A novel biphenotypic leukemia cell line, NALM-29, was established from a 46-year old Japanese male patient with acute lymphoblastic leukemia (ALL). The primary leukemic blasts showed a common ALL phenotype with CD19+, CD10+, CD13-, HLA-DR+ and Igs-. NALM-29 cells display biphenotypic characteristics: expression of the intracellular enzyme myeloperoxidase at the mRNA and protein level and cell surface positivity for CD19, CD10, CD13, CD33 and HLA-DR. NALM-29 fulfills EGIL criteria as B-cell precursor (BCP) leukemia B-II type. NALM-29 cells have a lymphoblastic morphological appearance; the immunoglobulin heavy chain gene is rearranged. NALM-29 cells responded significantly to the proliferative stimuli of FLT-3 ligand and IL-7, but not to GM-CSF, IL-3, IL-6, PIXY-321 or SCF. Proliferation of cells was inhibited significantly by IL-4, TNF-alpha or TNF-beta treatment. Cytogenetic analysis revealed the characteristic t(9;22)(q34;q11); expression of the m-bcr e1-a2 BCR-ABL fusion gene (typically found in ALL) was determined by PCR amplification of cDNA. The immunological, cytogenetic and functional characterization of NALM-29 suggests that this cell line may represent a scientifically significant in vitro model for BCP-type leukemia cells with biphenotypic characteristics. PMID- 10456672 TI - Effect of all-trans retinoic acid on chemotherapy induced apoptosis and down regulation of Bcl-2 in human myeloid leukaemia CD34 positive cells. AB - Acute myeloid leukaemia (AML) is a heterogeneous malignant disease in which disease progression at the level of CD34 positive cells has a major impact in drug resistance and relapse. The multi-drug resistance (MDR1) gene product, P glycoprotein is expressed mainly in CD34 positive AML cells and Bcl-2 is expressed simultaneously with several putative drug resistance parameters in these cells. Bcl-2 over-expression is associated with CD34 positivity, poor response to chemotherapy and reduced overall survival in AML patients. Recently, all-trans retinoic acid (RA) has been reported to enhance cytarabine-induced apoptosis and downregulate Bcl-2 in several human myeloid leukaemia CD34 negative cells. The two CD34 positive human myeloid leukaemia cell lines: KG1 and KGla have the unique feature of expressing significant functional P-glycoprotein. Thus, the efficacy of RA in enhancing cytrabine- and fludarabine-induced apoptosis and overcoming the resistance was examined in both KG1 (CD34+CD7-) and KGla (CD34+CD7+) human myeloid leukaemia cells in the present study. Both cytarabine and fludarabine induced a dose dependent increase in the number of apoptotic cells in both CD34 positive cell types. Interestingly, the cytarabine induced apoptosis was significantly more than fludarabine-induced apoptosis in both cell types. All-trans RA alone failed to induce apoptosis or inhibit proliferation of either of the two human CD34 positive leukaemia cell types. However, RA enhanced cytarabine- or fludarabine-induced apoptosis and inhibition of proliferation in KG1 CD34+CD7- but not in KGla CD34+CD7+ myeloid leukaemia cells. As single agents, RA, cytarabine and fludarabine reduced Bcl-2 expression in a dose dependent manner in both cell types. Using a quantitative ELISA assay, the Bcl-2 protein concentration was reduced by 86 or 100%, after 72 h of treatment with 10 microM cytarabine or fludarabine, respectively, in both CD34 positive leukaemia cell types. The addition of RA to cytarabine enhanced its induced reduction of Bcl-2 in KG1 CD34+CD7- but not in KGla CD34+CD7+ human myeloid leukaemia cells. Meanwhile, RA failed to augment fludarabine-induced reduction of Bcl-2 in both cell types. In conclusion, the present results suggest a potential role for the combination of RA and cytarabine in the treatment of refractory and/or relapsed AML patients with CD34+CD7- but not CD34+CD7+ blast cells. PMID- 10456673 TI - Prolactin blocks glucocorticoid induced cell death by inhibiting the disruption of the mitochondrial membrane. AB - Prolactin (PRL) has been reported to inhibit dexamethasone (Dex) induced cell death. Nevertheless, the mechanism through which PRL exerts its protective effect is still not unravelled. Here, we analyse the effect of PRL at different stages of the glucocorticoid (GC) apoptotic pathway in PRL dependent cells (Nb2 cells). PRL blocks completely the GC induced loss of the mitochondrial transmembrane potential (delta psi(m)) and consequently phosphatidylserine (PS) exposure and loss of DNA content. Although PRL promotes an upregulation of the bcl-2 expression, simultaneous addition of PRL to GC fails to maintain even the normal levels of this anti-apoptotic protein. This finding excludes a critical role for bcl-2 in the PRL protective effect against GC. GC induced delta psi(m) disruption can be inhibited by the ICE-like inhibitor zVAD-fmk but not by ICE inhibitor tetrapeptide acetyl-Tyr-Val-Ala-Asp.chloromethylketone (YVAD-cmk) nor by caspase 3 inhibitor zDEVD. It can be speculated that PRL blocks delta psi(m) disruption by inhibiting an unknown caspase activated by GC. PMID- 10456674 TI - Hairy cell leukemia with translocation (11;20)(q13;q11) and overexpression of cyclin D1. AB - We report on a male Japanese patient with hairy cell leukemia (HCL). A cytogenetic study with lipopolysaccharide stimuli showed a novel translocation (11;20)(q13;q11) in 10% of the analyzed cells. Northern blot analysis and RT-PCR analysis for cyclin D1 revealed the overexpression of cyclin D1, although the southern blot analysis of PRAD1 gene showed no rearrangement. In this particular case, the t(11;20)(q13;q11) might play some role in the oncogenesis of HCL and the overexpression of cyclin D1. PMID- 10456675 TI - Blast cell morphology in acute myeloid leukemia with trisomy 13. PMID- 10456676 TI - Occurrence of emodin, chrysophanol and physcion in vegetables, herbs and liquors. Genotoxicity and anti-genotoxicity of the anthraquinones and of the whole plants. AB - 1,8-Dihydroxyanthraquinones, present in laxatives, fungi imperfecti, Chinese herbs and possibly vegetables, are in debate as human carcinogens. We screened a variety of vegetables (cabbage lettuce, beans, peas), some herbs and herbal flavoured liquors for their content of the 'free' anthraquinones emodin, chrysophanol and physcion. For qualitative and quantitative analysis, reversed phase HPLC (RP-LC), gas chromatography-mass spectrometry (GC-MS) and RP-LC-MS were used. The vegetables showed a large batch-to-batch variability, from 0.04 to 3.6, 5.9 and 36 mg total anthraquinone per kg fresh weight in peas, cabbage lettuce, and beans, respectively. Physcion predominated in all vegetables. In the herbs grape vine leaves, couch grass root and plantain herb, anthraquinones were above the limit of detection. Contents ranged below 1 mg/kg (dry weight). All three anthraquinones were also found in seven of 11 herbal-flavoured liquors, in a range of 0.05 mg/kg to 7.6 mg/kg. The genotoxicity of the analysed anthraquinones was investigated in the comet assay, the micronucleus test and the mutation assay in mouse lymphoma L5178Y tk+/- cells. Emodin was genotoxic, whereas chrysophanol and physcion showed no effects. Complete vegetable extract on its own did not show any effect in the micronucleus test. A lettuce extract completely abolished the induction of micronuclei by the genotoxic anthraquinone danthron. Taking into consideration the measured concentrations of anthraquinones, estimated daily intakes, the genotoxic potency, as well as protective effects of the food matrix, the analysed constituents do not represent a high priority genotoxic risk in a balanced human diet. PMID- 10456677 TI - Reproductive effects of dietary soy in female Wistar rats. AB - This study was designed to assess the effects of administration of dietary soy on reproductive function and fertility of female Wistar rats. Four groups, each of 20 females, were used. Control animals were fed a standard cereal-based diet for rats. Treated animals were fed a standard diet supplemented with 0.7%, 1.2% or 2.4% of a soy extract. Treatment started at weaning and continued until day 7 post-partum (day of sacrifice). Growth depression was seen in the 2.4% soy group. Vaginal opening occurred earlier in females receiving soy supplemented feed when compared with controls. Analysis of vaginal smears revealed that all animals were cycling, although an increase in the mean duration of each cycle was seen in the 2.4% soy group. Uterine effects were observed in high-dose females and included increases in weight, oedema, endothelial hyperplasia and leucocytic infiltration. Vaginal modifications (i.e. inflammation, hyperkeratosis and dyskeratosis) and alterations in the distribution of follicular size in the ovaries were also observed among treated animals. These data suggest that long-term exposure to high doses of phytoestrogens can produce significant agonistic actions in several oestrogen-dependent tissues and parameters, even though in this model no clear influence on reproductive processes was observed. PMID- 10456678 TI - Toxicological and antioxidant effects of short-term dehydroepiandrosterone injection in young rats fed diets deficient or adequate in vitamin E. AB - This study examined the in vivo antioxidant and/or prooxidant effect of short term dehydroepiandrosterone (DHEA) injection and the effect of dietary vitamin E. Male Sprague-Dawley rats (4 wk old) were fed vitamin E-deficient or vitamin E adequate (30 mg DL-alpha-tocopheryl acetate/kg) diet for 4 weeks followed by intraperitoneal injection of DHEA for 1 week. The results showed that DHEA injection caused a dose-dependent decrease in body weight, and this effect was more pronounced in vitamin E-deficient rats. In contrast, DHEA injection significantly increased liver, kidney and adrenal weights. Hepatic vitamin E content was significantly lowered by vitamin E deficiency, which led to significantly increased ex vivo and iron-induced lipid peroxidation. DHEA injection did not affect hepatic vitamin E content but significantly decreased ex vivo and iron-induced lipid peroxidation in vitamin E-deficient rats. Hepatic total sulfhydryl (SH) groups and non-protein SH contents were not affected by vitamin E but were significantly increased by DHEA injection, which at 100 mg/kg was not more effective than at 50 mg/kg. Hepatic glutathione S-transferase (GST) activity was significantly decreased by DHEA, but vitamin E alleviated such a decrease. DHEA injection significantly increased hepatic glucose 6-phosphate dehydrogenase (G6PD) activity, and the effect was dose dependent in vitamin E deficient rats. Thus, DHEA may compensate for vitamin E deficiency in vivo, and this effect is masked when dietary vitamin E is adequate. The antioxidant effect of DHEA is accompanied by decreased body weights, enlarged (fat-laden) tissues and altered activities of hepatic GST and G6PD. PMID- 10456679 TI - Effect of oxidized fish oil and alpha-tocopherol on the peroxidation of erythrocyte membrane phospholipids in rats. AB - The effects of dietary oxidized fish oil and alpha-tocopherol on the thiobarbituric acid (TBA) values and phospholipid hydroperoxide levels of the erythrocyte membrane were studied in rats. No significant differences in the TBA values or phospholipid hydroperoxide levels of the membrane were observed between groups fed either oxidized fish oil or control diets. Furthermore, there were no marked differences in these values whether or not the groups were administered diets containing added alpha-tocopherol. These results suggest that the intake of oxidized fish oil and the supplementation with alpha-tocopherol do not influence the level of lipid peroxidation in the erythrocyte membrane. PMID- 10456680 TI - Dietary precursors of trimethylamine in man: a pilot study. AB - An increased urinary excretion of trimethylamine and its N-oxide were observed in man following the oral intake (15 mmol) of choline (63% dose as trimethylamine and its N-oxide), D,L-carnitine (31% dose) and trimethylamine N-oxide (78% dose). Similar ingestion of betaine, creatinine or lecithin failed to elicit any significant increases. Of 46 different foods investigated, only fish and other sea-products gave rise to significant increases in urinary trimethylamine and N oxide. Ingestion of fruits, vegetables, cereal and dairy produce, and meats had no measurable effects. Reasons for the apparent lack of trimethylamine provision by foods previously thought to be precursors are given and the role of gut microflora highlighted. PMID- 10456681 TI - Safety evaluation of phytosterol esters. Part 2. Subchronic 90-day oral toxicity study on phytosterol esters--a novel functional food. AB - Phytosterol esters (PE) are intended for use as a novel food ingredient, primarily in margarines and spreads as a functional component with plasma cholesterol lowering activity. Phytosterols and their esters are present naturally in vegetable oils and on average people consume 200 mg/day, but their consumption at this level is not sufficient to lower plasma cholesterol levels. Therefore, through the incorporation of PE into margarines/spreads, the intake can be increased by approximately 10-fold by consuming the PE-containing margarine/spread at normal intake levels. As part of an extensive programme of safety evaluation studies a subchronic rat toxicity study has been conducted in which groups of Alpk:AP(f)SD (Wistar derived) rats (20 males and 20 females/group) were fed diets containing PE at levels of 0, 0.16, 1.6, 3.2 and 8.1% (w/w) in the diet for 90 days. Throughout the study, clinical observations, body weights, and food and water consumption were measured. At the end of the study the rats were subjected to a full post-mortem examination, cardiac blood samples were taken for clinical pathology, selected organs were weighed, and a full tissue list was taken for subsequent histological examination. There were no treatment-related changes that were considered to be of toxicological significance. Therefore, a nominal PE concentration of 8.1% was considered to be the no-observed-adverse- effect level (NOAEL) following daily oral administration to rats for 90 days. This was equivalent to a dose of 6.6 g/kg body weight/day PE or 4.1 g/kg/day phytosterol. PMID- 10456682 TI - Effects of vomitoxin ingestion on murine models for systemic lupus erythematosus. AB - Dietary exposure to the trichothecene vomitoxin (VT) results in reduced body weight gain, elevated serum IgA, terminal differentiation of Peyer's patch B cells to IgA secreting plasma cells haematuria, and increased kidney mesangial IgA accumulation in B6C3F1 mice and other inbred strains. These effects closely mimic a human autoimmune-like kidney disease known as IgA nephropathy. Using NZBW/F1, MRL/lpr, and BXSB mouse strains as models of systemic lupus erythematosus, we assessed whether consumption of diet containing 5 ppm or 10 ppm VT will similarly affect mice genetically prone to autoimmunity. Reduced weight gains were seen in NZBW/F1 and MRL/lpr mice fed both doses of VT within 2-3 weeks. In contrast, VT had little effect on weight gain by BXSB mice. Serum Ig levels in all three strains generally did not differ from control mice. Haematuria was significantly increased when all three strains were fed VT. In NZBW/F1 Peyer's patch cultures stimulated with lipopolysaccharide (LPS), prior VT exposure significantly increased the IgG and IgM secretion but had no effect on IgA. In MRL/lpr Peyer's patch cultures stimulated with LPS, VT exposure increased IgA secretion but not IgM or IgG. BXSB Peyer's patch cultures prepared from VT treatment groups produced significantly more IgA than controls when cultured with LPS or Concanavalin A. Whereas mesangial deposition of IgA and IgG was significantly lower in the treatment groups of NZBW/F1 and MRL/lpr mice compared with control, BXSB mice had significantly higher IgA, IgG, and complement (C3) deposition when fed VT. The results suggest that although dietary VT differentially affected mice with aberrant immune systems, these strains did not appear to be any more sensitive to the mycotoxin than were more immunologically robust inbred strains. PMID- 10456684 TI - Thresholds in contact sensitization: theoretical and practical considerations. AB - The existence of thresholds for both the induction and the elicitation of contact sensitization is an important issue for hazard assessment in this area of toxicology. In this paper, the evidence for such thresholds has been reviewed from both theoretical and practical perspectives. When the mechanisms of skin sensitization are considered, the existence of thresholds can be shown to occur at several stages. They are both quantitative, for example in terms of the degree of protein haptenation and in the sufficiency of the signals for Langerhans cell migration and maturation, as well as qualitative, in terms of the type of immune response that is engaged. Such considerations are evidenced by a substantial body of practical observation. In humans and in animal models of skin sensitization there is abundant evidence for the existence of thresholds for both the induction and the elicitation of reactions. In addition to, and in distinction from, the experimental situation, in the general human population there is extensive evidence for threshold effects. This evidence arises directly from the observation that only a proportion of those who are exposed become sensitized (i.e. are patch test positive), and of that latter group only a proportion develop allergic contact dermatitis. PMID- 10456683 TI - Toxicological studies on procyanidin B-2 for external application as a hair growing agent. AB - Procyanidin B-2 [epicatechin-(4beta --> 8)-epicatechin] is one of condensed tannin that exists widely in plants. We have reported previously that procyanidin B-2 possesses hair epithelial cell growth-promoting activity and stimulates anagen induction in hair cycle progression. To evaluate the safety of topical procyanidin B-2 as a hair growing agent, we examined the mutagenicity, acute subcutaneous injection, primary irritation, skin sensitization, and eye irritation of this compound. Mutagenicity tests using bacteria showed procyanidin B-2 to be non-mutagenic. Chromosomal aberration tests using CHL cells indicated that procyanidin B-2 caused polyploidy but no structural aberrations. In micronucleus tests for mutagenicity using mice, procyanidin B-2 was negative. Acute subcutaneous injection study using rats revealed no symptoms of significant injury. The lethal dose of procyanidin B-2 is greater than 2000 mg/kg (subcutaneous injection). Primary irritation tests using rabbits indicated that procyanidin B-2 containing preparation shows no primary irritation. In the guinea pig maximization test, there was no evidence of sensitization to procyanidin B-2. In primary ocular irritation tests using rabbits, procyanidin B-2 containing preparation and vehicle showed slight irritation of conjunctivae which is assumed to be caused by ethanol. It is suggested that topical procyanidin B-2 is safe and acceptable from the series of toxicological tests. PMID- 10456685 TI - Muscle necrosis in rats induced by 2-methoxy-p-phenylenediamine. AB - p-Phenylenediamine, together with several of its amino and alkyl derivatives, are known to be myotoxic in animals and man. In the present study, it was found that 2-methoxy-p-phenylenediamine, a component of oxidative hair dyes, similarly causes necrosis of skeletal muscle (gastrocnemius, diaphragm and tongue) in rats. The myotoxicity of 2-methoxy-p-phenylenediamine was much greater than that of p phenylenediamine itself. This observation is consistent with the relative efficacy of these substances in disrupting mitochondrial metabolism, an effect which is believed to be responsible for the selective toxicity of p phenylenediamine derivatives to muscle. PMID- 10456686 TI - A limited developmental toxicity study of pentane by inhalation in the rat. AB - Pentane (CAS No. 109-66-0) is a widely-used chemical that finds many industrial applications. As part of a program looking at the inhalation toxicity of pentane, we studied the potential effects of the chemical on the developing embryo. This communication reports the findings of a limited study which was used to determine whether a full-scale developmental toxicity study was necessary. Groups of 7 to 8 pregnant rats were exposed by inhalation to pentane, 6 hr/day from gestation days 6 through 15 at concentrations of 0 (control), 1000, 3000 and 10,000 ppm, Maternal body weights, clinical signs and food consumption were measured; foetuses were weighed and examined for gross external development. Skeletal and internal organ evaluations were not performed. There were no effects in either the maternal or fetal rats (at concentrations up to and including 10,000 ppm) that could be associated with pentane exposure. Based on those findings, we did not conduct a full-spectrum developmental toxicity study in rats, but concluded that it is unlikely that the foetus would be adversely affected by pentane exposure. PMID- 10456687 TI - Differential gene expression in drug metabolism and toxicology: practicalities, problems and potential. AB - 1. An important feature of the work of many molecular biologists is identifying which genes are switched on and off in a cell under different environmental conditions or subsequent to xenobiotic challenge. Such information has many uses, including the deciphering of molecular pathways and facilitating the development of new experimental and diagnostic procedures. However, the student of gene hunting should be forgiven for perhaps becoming confused by the mountain of information available as there appears to be almost as many methods of discovering differentially expressed genes as there are research groups using the technique. 2. The aim of this review was to clarify the main methods of differential gene expression analysis and the mechanistic principles underlying them. Also included is a discussion on some of the practical aspects of using this technique. Emphasis is placed on the so-called 'open' systems, which require no prior knowledge of the genes contained within the study model. Whilst these will eventually be replaced by 'closed' systems in the study of human, mouse and other commonly studied laboratory animals, they will remain a powerful tool for those examining less fashionable models. 3. The use of suppression-PCR subtractive hybridization is exemplified in the identification of up- and down regulated genes in rat liver following exposure to phenobarbital, a well-known inducer of the drug metabolizing enzymes. 4. Differential gene display provides a coherent platform for building libraries and microchip arrays of 'gene fingerprints' characteristic of known enzyme inducers and xenobiotic toxicants, which may be interrogated subsequently for the identification and characterization of xenobiotics of unknown biological properties. PMID- 10456688 TI - Quantitative immunohistochemical analysis of the glutathione S-transferase GSTM1: in situ phenotyping in archival material. AB - 1. GSTM1 is present in only approximately 50% of Caucasian individuals and deficiency of GSTM1 is associated with susceptibility to a growing number of diseases, especially cancer. Thus, a method that would allow accurate, retrospective determination of the GSTM1 phenotype in different patient populations would have many applications. 2. Developed, therefore, is a quantitative, image-analysis-based immunohistochemical technique for the analysis of GSTM1 protein in paraffin-embedded tissue samples. It was applied to the determination of the GSTM1 phenotype using liver biopsies taken from 70 patients. 3. Of the 70 cases (depending on the cut-off point), 51-54% were deficient in GSTM1. A single 27 kD band characteristic for GSTM1 was found in seven of 16 cases analysed by Western blotting using the same GSTM1 antibody as in the immunohistochemical analysis. There was a good correlation (r = 0.87) between the staining intensity of the GSTM1 band and the staining intensity evaluated by immunohistochemistry. 4. It is concluded that this quantitative immunohistochemical method permits accurate determination of the GSTM1 phenotype and is well suited for retrospective analysis of GSTM1 expression in specific tissues in situ. PMID- 10456689 TI - Metabolism of artelinic acid to dihydroqinqhaosu by human liver cytochrome P4503A. AB - 1. Artelinic acid (AL), a water-soluble artemisinin analogue for treatment of multidrug resistant malaria, is metabolized to the active metabolite dihydroqinghaosu (DQHS) solely by CYP3A4/5. Although AL is not metabolized by CYP2C9, it does inhibit diclofenac 4-hydroxylase activity with an IC50 = 115 microM. Interestingly, AL activates CYP2D6-mediated bufuralol metabolism in human liver microsomes but not recombinant CYP2D6-Val by approximately 30% at AL concentrations up to 100 microM. 2. In human liver microsomes, AL is metabolized to DQHS with a Km = 157 +/- 44 microM and Vmax = 0.77 +/- 0.56 nmol DQHS/min/mg protein. Human recombinant CYP3A4 catalysed the conversion of AL to DQHS with a Km = 102 +/- 23 microM and a Vmax = 1.96 +/- 0.38 nmol DQHS/min/nmol P450. The kinetic parameters (Km and Vmax) for DQHS formation from CYP3A5 were 189 +/- 19 microM and 3.60 +/- 0.42 nmol DQHS/min/nmol P450 respectively. 3. Inhibition studies suggest that azole antifungals and calcium channel blockers may present clinically significant drug drug interactions. In human liver microsomes, ketoconazole and miconazole were potent competitive inhibitors of DQHS formation with a Ki = 0.028 and 0.124 microM respectively. Verapamil is a non-competitive inhibitor of DQHS formation in human liver microsomes with a Ki = 15 microM. PMID- 10456690 TI - Involvement of CYP2D6 in the in vitro metabolism of amphetamine, two N alkylamphetamines and their 4-methoxylated derivatives. AB - 1. Amphetamine (AM) and five amphetamine derivatives, N-ethylamphetamine (NEA), N butylamphetamine (NBA), 4-methoxyamphetamine (M-AM), 4-methoxy-N-ethylamphetamine (M-NEA) and 4-methoxy-N-butylamphetamine (M-NBA) were incubated with microsomal preparations from cells expressing human CYP2D6 to determine whether the enzyme was capable of catalyzing the direct ring oxidation of all substrates; the N dealkylation of NEA, NBA, M-NEA and M-NBA; and the O-demethylation of M-AM, M-NEA and M-NBA. 2. None of the six compounds examined was N-dealkylated to any extent. 3. The only metabolites produced from AM, NEA and NBA were the corresponding ring 4-hydroxylated compounds, and the rates of formation were low. 4. All ring 4 methoxylated substrates were efficiently O-demethylated by CYP2D6 to their corresponding phenols. The size of the N-alkyl group influenced the rates of formation of these phenolamines. In contrast to reported findings with 2- and 3 methoxyamphetamines, none of the 4-methoxyamphetamines was ring-oxidized in the CYP2D6 enzyme system to 2- or 3-hydroxy-4-methoxyamphetamines or to dihydroxyamphetamines. PMID- 10456692 TI - Metabolism and disposition of GTS-21, a novel drug for Alzheimer's disease. AB - 1. GTS-21, a novel drug for Alzheimer's disease, is currently under clinical development. In the current study, the metabolism and disposition of GTS-21 have been evaluated in rat and dog after single oral and intravenous administration. 2. Following oral administration of [14C]GTS-21 to rat, radioactivity was primarily excreted in the faeces (67%) via the bile with possible enterohepatic circulation. Urinary excretion of radioactivity in rat and dog was 20 and 19% respectively. 3. GTS-21 was rapidly and extensively absorbed after oral administration and rapidly cleared from plasma. The maximum concentration ratio of GTS-21 to total radioactivity in plasma was low, indicating first-pass or pre systemic biotransformation. 4. In rat, GTS-21 showed linear pharmacokinetics over doses ranging from 1 to 10 mg/kg with an absolute bioavailability of 23%. In dog, the absolute bioavailability was 27% at an oral dose of 3 mg/kg. 5. GTS-21 was O demethylated to yield compounds that were then subject to glucuronidation. Three of the metabolites in rat urine were isolated and characterized as 4-OH-GTS-21, 4 OH-GTS-21 glucuronide and 2-OH-GTS-21 glucuronide. The major urinary metabolites were 4-OH-GTS-21 glucuronide and 2-OH-GTS-21 glucuronide. 6. In vitro chemical inhibition of cytochrome P450 in human liver microsomes indicated that CYPIA2 and CYP2E1 were the isoforms primarily responsible for the O-demethylation of GTS-21, with some contribution from CYP3A. PMID- 10456693 TI - Surgical Internet at a glance: the Virtual Hospital. PMID- 10456691 TI - Biotransformation of protriptyline by filamentous fungi and yeasts. AB - 1. The potential of various fungi to metabolize protriptyline (an extensively used antidepressant) was studied to investigate similarities between mammalian and microbial metabolism. 2. Metabolites produced by each organism were isolated by high-pressure liquid chromatography and identified by nuclear magnetic resonance and mass spectrometry. The metabolites identified in one or more fungi were 2-hydroxyprotriptyline, N-desmethylprotriptyline, N-acetylprotriptyline, N acetoxyprotriptyline, 14-oxo-N-desmethylprotriptyline, 2-hydroxy acetoxyprotriptyline and 3-(5-hydrodibenzo[bf][7]annulen-5-yl)propanoic acid. 3. Among 27 filamentous fungi and yeast species screened, Fusarium oxysporum f. sp. pini 2380 metabolized 97% of the protriptyline added. Several other fungi screened gave significant metabolism of protriptyline, including Cunninghamella echinulata ATCC 42616 (67%), C. elegans ATCC 9245 (17%), C. elegans ATCC 36112 (22%), C. phaeospora ATCC 22110 (50%), F. moniliforme MRC-826 (33%) and F. solani 3179 (12%). 4. F. oxysporum f. sp. pini produced phase I and phase II metabolites and thus is a suitable microbial model for protriptyline metabolism. PMID- 10456694 TI - Flexner and the whole-time system: the second Flexner report and the whole-time system in American academic surgery. AB - BACKGROUND: Reform in medical education started in the mid 19th century and continued through the early decades of the 20th century. Both Carnegie and Rockefeller foundation monies were applied to the process of attempting to improve medical education. The University of Cincinnati College of Medicine, associated with a municipal hospital and a municipally owned university, offered a model for improvements in medical education in other municipal hospitals. The attempt to institute the "whole-time" ("full-time" salaried, "University") system, and restructure the University of Cincinnati College of Medicine along radical lines hitherto unknown in much of the country and in particular to the city of Cincinnati, disturbed the relationship of the College of Medicine with the community and echoes to this day. The tensions between the "University salaried" professors of clinical surgery and the practitioners attempting to practice at the institution have abated but remain unresolved. CONCLUSION: The history of the establishment of the full-time chairs, including the Christian R. Holmes Chair of Surgery, led to a rethinking of the role that the full-time system might play in American medical education. PMID- 10456695 TI - Retrospective analysis of the adequacy of oral contrast material for computed tomography scans in trauma patients. AB - BACKGROUND: This study examined the adequacy of oral contrast (OC) in adult trauma patients, the differences in the incidence of pathology and other patient characteristics related to OC adequacy, and the incidence of OC-associated emesis. METHODS: The cases of 129 adult trauma patients were reviewed from January to December 1996 for utilization of OC, administration time, time to computed tomography (CT), and emesis within 4 hours. CT films were reviewed for adequacy of OC (AC = adequate contrast, IC = inadequate contrast) and pathology. RESULTS: Data are presented as median and range, or percentage and 95% confidence interval (CI). Pathology was found in 24% (CI 17% to 31%) and was not associated with AC. OC administration to CT was 14 minutes (0 to 139). IC was present for 60% (CI 52% to 69%). Subjects with IC had higher injury severity scores. 10% (CI 5% to 15%) with OC had emesis. CONCLUSION: OC is not necessary for diagnostic accuracy, and may contribute to patient morbidity or discomfort. PMID- 10456696 TI - Fundoplication provides effective and durable symptom relief in patients with Barrett's esophagus. AB - BACKGROUND: Columnar-lined esophagus with intestinal metaplasia (IM), also called Barrett's esophagus, is a manifestation of severe gastroesophageal reflux (GER) and may predict poor symptom relief and high failure rate after fundoplication. We compared symptom scores and reoperation rates in GER patients with and without Barrett's esophagus. METHODS: Between July 1992 and July 1997, 646 patients underwent fundoplication (626 laparoscopic). Of 150 endoscopic biopsies of suspected columnar-lined esophagus, 80 confirmed IM, 50 identified cardiac or fundic epithelium, and 20 revealed only esophagitis. Typical GER symptoms were scored by patients preoperatively and postoperatively (0 to 4 scale). We compared symptom response (Wilcoxon rank sum test) and failure rates (t test) in patients with IM and GER controls without IM. Preoperative data were available for 74 IM patients and 496 controls. One-year follow-up was available in 45 IM patients and 301 controls. Intermediate follow-up (2 to 5 years) was available in 20 IM patients and 99 controls. RESULTS: Preoperatively and postoperatively, patients with IM reported heartburn, regurgitation, and dysphagia scores similar to controls. Procedure failure, requiring redo fundoplication, appeared more likely in IM patients than controls (6.3% versus 2.5%), but this difference did not reach statistical significance (P = 0.061). CONCLUSION: Fundoplication provides equivalent symptom relief for patients with and without IM. PMID- 10456697 TI - Small intestine protection from radiation by means of a removable adapted prosthesis. AB - BACKGROUND: A prosthesis has been designed to protect intestinal loops from radiation when postsurgical radiotherapy is necessary in cancer treatment. It is a silicone balloon that allows the small bowel to be pushed back away from the radiation field, and it is easily removed at the conclusion of radiotherapy. METHODS: The device was used in 22 patients: 5 retroperitoneal tumors and 17 pelvic cancers. After surgical resection of the tumor, the device is placed either in the retroperitoneal area or in the pelvic cavity. A polyglactine 910 mesh is placed between the spacer and the bowel to prevent incarceration of the loops. The prosthesis can be filled or emptied between each radiation course and finally removed by means of a short incision under local or locoregional anesthesia. RESULTS: The tolerance of the small intestine to radiation therapy has been satisfactory in each case, with a mean follow-up of 24.5 months (range 10 to 73). No modification of biological parameters was observed during the pelvic radiation therapy at 30, 45, and 65 Gy. CONCLUSION: This device should appears to efficient for prevention of bowel injury during postsurgical radiation in successful treatment of abdominal, pelvic, or retroperitoneal tumors when indicated. PMID- 10456698 TI - Hyperthermic isolated regional perfusion for the treatment of osteosarcoma in the lower extremity. AB - BACKGROUND: In order to reduce the systemic toxity of cisplatin and to increase the effect on the local tumor, facilitating limb-preserving procedures, we have been using hyperthermic isolated regional perfusion (HIRP) for the treatment of osteosarcoma in addition to preoperative chemotherapy. MATERIALS AND METHODS: Twenty patients with an osteosarcoma in a lower extremity were treated with both preoperative chemotherapy and HIRP. RESULTS: The chemotherapeutic effects of preoperative chemotherapy with HIRP was grade III in 6, and grade IV (no viable cell areas observed) in 7. The surgical margins could be reduced in these 13 cases of good responders showing over 90% necrosis (grade III or grade IV), which mainly involved postnecrotic fibrosis and osteosclerosis. CONCLUSION: HIRP can help to control the tumor locally. A more conservative operation was then possible in cases of good responders to preoperative chemotherapy with HIRP. PMID- 10456699 TI - Preliminary prospective randomized experience with vascular clips in the creation of arteriovenous fistulae for hemodialysis. AB - BACKGROUND: A new vascular clipping system (VCS) has been introduced for clinical use. This study was developed to determine whether patency rates and treatment outcomes for anastomoses in hemodialysis access surgery can be improved by the application of this new technology. METHODS: During a 10-month period, 96 consecutive patients requiring autologous fistula, synthetic fistula, or graft revisions for hemodialysis were prospectively randomized into two treatment groups. Anastomoses were constructed with the VCS in 46 patients and with polytetrafluoroethylene (PTFE) suturing in 49 patients. The procedures were performed in an outpatient setting, at a large medical center, by one surgeon. Time to construct the anastomosis, amount of bleeding from the anastomotic site, and primary and secondary graft patencies were analyzed. RESULTS: The mean time to construct anastomoses in autologous and synthetic grafts using VCS clips was significantly shorter than with sutures (autologous 14 versus 22 minutes, P = 0.0001; and new grafts 26 versus 30 minutes, P = 0.04). Blood loss was less in anastomoses done with the VCS clip; however, statistical significance was reached only for autologous fistulas (P = 0.0001). At 2-year follow-up, primary and secondary patencies were similar for both the VCS and suture groups with the exception that a statistically significant improvement was found in secondary patency for autologous fistulas when performed with the VCS. CONCLUSIONS: The VCS clip is easy to use and produces a more rapid anastomosis with less bleeding. Primary and secondary patencies using the VCS clip were equivalent or improved when compared with standard suture. In our experience, no complications have been attributed to the use of the clip technique. PMID- 10456700 TI - Silicone venous access devices positioned with their tips high in the superior vena cava are more likely to malfunction. AB - BACKGROUND: Silicone venous access devices (VADs) are commonly used for multiple intravenous infusions and blood sampling in chronically ill patients, particularly cancer patients. These devices are susceptible to malfunctions most commonly characterized by difficulty infusing fluids or withdrawing blood. We hypothesized that the incidence of such malfunctions is primarily related to the position of the catheter tip relative to the superior vena cava/right atrial junction. METHODS: We reviewed medical records for 141 patients in whom silicone venous access devices were used. Chest radiographs obtained following catheter insertion were reviewed by a single radiologist to rule out pneumothorax and determine the position of the catheter tip. Other potential factors included in the analysis were catheter brand, the vein into which the catheter was inserted, and the side (right or left). A malfunction was defined as resistance to flushing or infusion of a substance, complete inability to infuse a substance, resistance to blood aspiration attempts, or absence of blood return with aspiration, occurring any time during which the catheter was in place. Multiple logistic regression was used to determine which factor(s) most strongly predicted catheter malfunction. RESULTS: Catheter tip location was the only factor that was statistically predictive of malfunctions (coefficient 0.842, P < 0.001). A significant increase in malfunctions was observed in cases where the catheter tip was located greater than 4 cm superior to the junction of the right atrium and the superior vena cava (z-test of proportions, P = 0.003). Malfunctions were minimized in those cases where the catheter tip was located in the right atrium. CONCLUSIONS: These results indicate that malfunctions can be minimized in silicone venous access catheters by locating the catheter tip as close to the superior vena cava/right atrial junction as possible, or slightly inside the right atrium. To ensure proper location of the catheter tip, placement should be performed under fluoroscopy, and a radiograph should be obtained immediately following placement, with the patient in the upright position. PMID- 10456701 TI - Is splenectomy for massive splenomegaly safe in children? AB - PURPOSE: To study and analyze the causes, etiology, morbidity, mortality and therapeutic value of splenectomy performed for massive splenomegaly in children. METHODS: The medical records of 115 children less than 18 years old who had splenectomy for various hematological disorders were reviewed. Twenty of them had splenectomy for massive splenomegaly (spleen weight > or =1,000 g). The records of these were reviewed for age at operation, gender, hematological diagnosis, indication for splenectomy, operative procedures, postoperative complications, and outcome. RESULTS: Twenty children had splenectomy for massive splenomegaly. There were 16 males and 4 females. Their ages ranged from 4 to 15 years (mean 11.2). Twelve had sickle cell disease, 5 had sickle-beta-thalassemia, 1 had beta thalassemia major, 1 had thalassemia intermediate, and 1 had chronic myeloid leukemia. The indications for splenectomy were hypersplenism in 11, recurrent splenic sequestration crisis in 8, and splenic abscess in 1. The transfusion requirements in the patient with beta-thalassemia major decreased markedly postoperatively from 18 transfusions/year to only 4 transfusions/year; and for those with hypersplenism, there was a marked improvement in their blood parameters following splenectomy. The patient with thalassemia intermediate required no more blood transfusions. There was no mortality. The immediate postoperative morbidity was 10% for those with massive splenomegaly compared with 6.3% for those with splenomegaly <1,000 g. CONCLUSIONS: With good perioperative management, splenectomy in children with massive splenomegaly is both safe and effective. PMID- 10456702 TI - Selective endoscopic retrograde cholangiography prior to laparoscopic cholecystectomy for gallstones. AB - BACKGROUND: To assess the outcome of endoscopic retrograde cholangiography (ERC) before laparoscopic cholecystectomy (LC) for symptomatic gallbladder and suspected duct stones. METHODS: During 3 years, one or more of four criteria led to ERC: jaundice, choledocus >8 mm, cholestasis, and severe biliary pancreatitis. Endoscopic extraction (ESE) of ductal stones was attempted before LC. RESULTS: In all, 990 patients were prospectively included. There were no exclusions. There were no deaths. A multivariate logistic regression analysis identified jaundice (P = 0.001), pancreatitis (P = 0.001), and cholestasis (P = 0.001) as statistically significant predictors of ductal stones. Choledocus >8 mm was not a significant predictor (P = 0.12). A total of 155 (16%) patients underwent ERC for suspected stones: 21 of 155 (13%) patients had no stones; and 6 of 134 (4%) patients had stone impaction cleared at open surgery. ERC clearance rate was 95% (128 of 134). LC was performed in 149 of 155 patients after a median interval of 3 days (range 1 to 7). Morbidity rates were 3% (4 of 134), 2% (3 of 149), and nil (0 of 6) after ESE, LC, or open surgery, respectively. Median hospital stay was 11 days. A total of 835 patients underwent LC with a 1.5% complication rate. Laparoscopic fluoro-cholangiography showed < or =3 mm-sized stones in 10 of 835 (1.2%) patients. No stones were reported at a median follow-up of 4 months including 990 patients. CONCLUSIONS: Ninety-five percent of patients with ductal stones can be successfully and safely managed by ERC prior to LC. PMID- 10456703 TI - A case-controlled study of laparoscopic compared with open lateral adrenalectomy. AB - BACKGROUND: Few studies have been done regarding laparoscopic transperitoneal lateral adrenalectomy compared with open transretroperitoneal lateral adrenalectomy in a case-controlled fashion. METHODS: A case-controlled study of 40 laparoscopic and 40 open adrenalectomies was done in patients who were matched for age, gender, endocrine disorder, side and size of tumor, and area of body surface. Follow-up was complete in 92.5% of the patients, with a mean follow-up period of 30 months. RESULTS: Statistically significant differences (P <0.05) were present (laparoscopic versus open) when the following results were compared: estimated blood loss (40 g versus 172 g), operating time (147 versus 79 minutes), analgesic equivalents (2.9 versus 5.2 times), hospital stay (12 versus 18 days), and late morbidity (0% versus 47.5%). There were no statistically significant differences between the laparoscopic and open groups with regard to time to oral intake, time to walking, intraoperative and early complications, and total cost. CONCLUSIONS: Laparoscopic adrenalectomy is a safe technique that results in greater patient comfort, decrease in estimated blood loss, and earlier discharge than open adrenalectomy, with no increase in cost. It should be adopted as the technique of choice for the removal of functioning adenomas and for adrenal masses less than 6 cm in diameter. PMID- 10456704 TI - Surgical technique for inferior mesenteric vein to renal vein shunt in portal hypertension. AB - An important role still exists for the creation of surgical portasystemic shunts. Multiple techniques have been described. However, no particular one is satisfactory for all clinical situations. The objective of the present paper is to describe an alternative surgical technique for the creation of a decompressive portasystemic shunt. This technique consists of an end-to-side anastomosis between the inferior mesenteric vein and the left renal vein. In our experience, the inferior mesenteric vein to left renal vein anastomosis may be a useful portal decompressive shunt for patients with otherwise difficult peri-portal or peri-pancreatic anatomic exposures. This technique should be a useful tool in the armamentarium of surgeons dealing with patients who need portal decompressive surgical shunts. PMID- 10456705 TI - Dissemination of melanoma cells within electrocautery plume. AB - BACKGROUND: The observed occurrence of port site recurrence in laparoscopic surgery for malignant disease has stimulated interest in the dissemination of tumor cells during surgery. Study of electrocautery smoke has revealed the presence of large particles and viable viruses. The purpose of this study was to determine if viable malignant cells are present in suspension within the electrocautery plume. METHODS: Pellets of B16-F0 mouse melanoma cells were cauterized and the plume collected into culture medium. In part 1 of this study, the trypan blue assay was used to assess cell viability immediately after collection and 7 days later. A cautery current of 30 W was applied for 5 minutes. In part 2, the tetrazolium (MTT) viability assay was used to assess cell viability after cauterization of tumor pellets at 10, 20, and 30 W for 5 seconds. RESULTS: Although intact melanoma cells were identified with the trypan blue assay immediately after plume collection, no viable cells were seen at 7 days using this assay. In part 2, viable melanoma cells were present in the culture wells at 7 days. Lower fulguration currents appeared to yield higher cell counts: 2,250 cells/well at 10 W, 2,100 cells/well at 20 W, and 1,800 cells/well at 30 W. CONCLUSIONS: Results of this study confirm that application of electrocautery to a pellet of melanoma cells releases these cells into the plume. These cells are viable and may be grown in culture. This release of malignant cells may explain the appearance of port metastases at sites that are remote from the surgical dissection or that were never in direct contact with the tumor. PMID- 10456706 TI - Indications for and results of surgical therapy for male gynecomastia. AB - BACKGROUND: The objective of our study was to analyze factors determining diagnostic versus cosmetic indication and postoperative results in the treatment of gynecomastia. PATIENTS AND METHODS: Data from 100 patients and 141 breasts were analyzed retrospectively, and reevaluated by questionnaire (n = 81) and clinical examination (n = 33). Except for 2 patients, all underwent subcutaneous mastectomy through various incisions. RESULTS: Diagnostic surgery was exclusively performed in unilateral, nodular gynecomastia being preferentially of grade I. Higher grade, bilateral gynecomastia led mainly to cosmetic surgery. Minor complications (skin retraction, hypertrophic scars, hypesthesia, skin redundancy) occurred in 53% of patients and significantly more often in grade III or II gynecomastia. Each incision was preferentially associated with specific sequelae. However, 86% of patients were satisfied with surgical results. CONCLUSIONS: Laterality, consistency, grade, and age at onset of symptoms determine surgical indication. Despite the high number of sequelae due to preoperative grade and selected incision, most patients are satisfied with postoperative results. PMID- 10456707 TI - An endothelin receptor antagonist ameliorates injuries of sinusoid lining cells in porcine liver transplantation. AB - BACKGROUND: TAK-044 is an endothelin receptor antagonist. Whether the agent has protective effects on liver graft injuries from non-heart-beating donors is unknown. METHODS: In donor pigs, cardiac arrest was induced by stopping the respirator. Forty-five minutes after cessation of the respirator, the liver was flushed with University of Wisconsin (UW) solution, preserved for 8 hours at 4 degrees C, and transplanted orthotopically. The pigs were divided into two groups: a control group and a drug-treated group in which TAK-044 was given in the UW solutions (10 mg/L) and was administered to recipients (10 mg/kg body weight). RESULTS: TAK-044 treatment significantly increased recipient survival rate. After reperfusion of the graft, portal venous pressure and 15-minute retention rate of indocyanine green were significantly reduced in the drug treated group. Electron microscopic findings indicated that TAK-044 attenuated endothelial cell injuries. CONCLUSION: TAK-044 treatment improves the viability of livers harvested from non-heart-beating donors. The main effect of the agent is protection of endothelial cells from ischemia/reperfusion injuries. PMID- 10456708 TI - The association between telomerase, histopathological parameters, and KI-67 expression in breast cancer. AB - BACKGROUND: Telomerase is a ribonucleoprotein enzyme that appears to play an important role in carcinogenesis. Telomerase reactivation seems to be associated with immortalization and malignancy. METHODS: Using a polymerase chain reaction (PCR)-based assay known as the TRAP (telomeric repeat and amplification protocol) assay, we examined telomerase activity in 60 breast specimens prospectively collected from 39 patients undergoing elective breast surgery in our center. The specimens included adjacent noncancerous breast (n = 21), benign breast disease (n = 5), and infiltrating carcinoma (n = 34). Ki-67 expression was determined in 32 invasive breast cancer specimens using immunohistochemistry techniques. The histopathological features were determined by light microscopy by an experienced breast pathologist. RESULTS: Telomerase activity was detected in 24 (71%) of 34 infiltrating carcinomas. None of the adjacent noncancerous specimens nor the benign breast lesions expressed telomerase activity. Telomerase reactivation was significantly associated with nodal metastasis and Ki-67 expression. There was no significant association between telomerase activity and menopausal status, tumor grade, or tumor size. CONCLUSIONS: Telomerase reactivation is associated with the acquisition of malignancy in the human breast. Telomerase activity is significantly associated with nodal metastasis and cellular proliferation as measured by Ki-67 expression in human breast cancer. PMID- 10456709 TI - Supraomohyoid neck dissection in cancer of the oral cavity. AB - BACKGROUND: In oral cavity cancer, supraomohyoid neck dissection (SOHND) is becoming more popular for patients with N0 and N1 disease in the neck. The aim of this study was to assess the value of this surgical procedure. METHODS: The study included 237 previously untreated patients with oral cavity cancer. The neck treatment consisted of SOHND or functional neck dissection (FND). One hundred sixty patients underwent postoperative radiation therapy. Survival probabilities, neck recurrences, and distant metastases were analyzed according to the surgical procedure. RESULTS: For patients having undergone SOHND, the 5-year survival probabilities were 70.2% and 76.5% in N0 and N1 necks, respectively. The neck recurrence rate in SOHND was 2%. CONCLUSIONS: SOHND is an effective method of treatment for the clinically negative neck in patients with squamous cell carcinoma of the oral cavity. It also proves efficient, in conjunction with postoperative radiotherapy, for control of neck metastases in selected patients. PMID- 10456710 TI - Echocardiography in infective endocarditis. AB - BACKGROUND: Echocardiographic demonstration of valvular infection now ranks with positive blood cultures as one of the two major clinical criteria for diagnosis of infective endocarditis (IE), according to new, more accurate guidelines for diagnosis. Because early detection of IE and its complications is essential for determining whether to pursue medical therapy or to intervene surgically, transthoracic echocardiography (TTE) is an essential part of the initial examination of patients with suspected IE. METHODS: Using MEDLINE, we searched and reviewed all articles with the key words infective endocarditis and transesophageal echocardiography. RESULTS: With its superior imaging, transesophageal echocardiography (TEE) has proven to be more sensitive than TTE for the diagnosis of IE as well as in the detection of IE-associated complications. CONCLUSIONS: While superior in predicting which patients with IE have perivalvular abscess or prosthetic valve dysfunction and which are most susceptible to systemic embolism, TEE is more invasive and must be used selectively. PMID- 10456711 TI - The third heart sound. AB - BACKGROUND: In the current era of cost-effective delivery of health care, it is particularly timely to carefully reevaluate the clinical utility of selected physical signs. The third heart sound (S3) is one such sign that is the focus of the current review. METHODS: I conducted a computerized MEDLINE search of articles related to S3 published since 1966. For information before 1966, textbook and other cross-references from MEDLINE-identified sources were used. RESULTS: The presence of S3 may be the earliest clue to the presence of left ventricular failure. It predicts a high risk of postoperative complications in the setting of noncardiac surgery and is a predictor of response to digoxin in patients with congestive heart failure. However, the interobserver variation in its detection is high even among experienced clinicians. CONCLUSIONS: While the presence of S3 may have important clinical implications, high interobserver variation in detection limits its use. PMID- 10456712 TI - Epidemic hysteria in Virginia: the case of the phantom gasser of 1933-1934. AB - OBJECTIVE: We studied an example of epidemic hysteria occurring outside a closed community and involving fear of being "gassed." The description presented is that of a previously unrecorded case of epidemic hysteria in the state of Virginia during 1933-1934. DATA SOURCES: Data were gathered from contemporary newspaper accounts. CONCLUSION: The case of the Virginia "gasser" is one in a long series of epidemic hysteria incidents during the 20th century, coinciding with heightened awareness of environmental pollution and triggered by imaginary or exaggerated contamination threats. A recommendation is provided on how physicians should approach such episodes. PMID- 10456713 TI - Necrotizing fasciitis and malpractice claims. AB - BACKGROUND: Necrotizing fasciitis (NF) is an aggressive bacterial infection of the superficial fascia and subcutaneous tissues that is increasing in incidence. The high toll exacted by this illness provides a setting for malpractice claims. METHOD: We reviewed 180 consecutive malpractice claims submitted by attorneys for medical expert review between 1987 and late 1997. Four cases involved NF. RESULTS: Alleged failure to obtain timely surgical consultation was the basis for three claims, and alleged failure to prevent NF by proper nursing care was the basis for the fourth. Three cases were closed and one was settled. CONCLUSIONS: The cornerstone of risk management for a clinical presentation compatible with NF is immediate surgical consultation, with other diagnostic steps a secondary consideration. PMID- 10456714 TI - Factors influencing morning report case presentations. AB - BACKGROUND: One of the most widely accepted didactic conferences among residency training programs is morning report. This study examines several factors that may influence the content of case presentations at morning report. METHODS: This prospective survey is of cases presented over a 1-year period at two morning reports--a university hospital and its affiliated Veterans Affairs (VA) hospital. RESULTS: Of 286 morning reports, 227 questionnaires (79%) were completed. The most common subspecialty categories represented in the cases were cardiology (20.3% of cases), infectious disease (13.2%), gastroenterology (11.5%), pulmonary (11.0%), hematology (10.1%), and general medicine (6.2%). The case mix was not significantly different by type of hospital nor by chief resident. The discussion was inpatient oriented in 88.6% of cases. CONCLUSION: Morning report case mix was similar at a university and its affiliated VA hospital and was predominantly inpatient oriented. PMID- 10456715 TI - Irritable bowel syndrome and lactose maldigestion in recurrent abdominal pain in childhood. AB - BACKGROUND: The aim of this study was to evaluate the impact of irritable bowel syndrome (IBS) and lactose maldigestion in children with recurrent abdominal pain. METHODS: Children who had abdominal pain associated with defecation or change in bowel habit, disordered defecation, and distension were diagnosed with IBS, and lactose maldigestion was defined by lactose breath hydrogen testing. Children with IBS were managed with increased fiber intake, while those with lactose maldigestion restricted dietary lactose. A telephone survey was conducted to determine the response to treatment. RESULTS: The mean age of the 59 boys and 87 girls was 9.5 +/- 3.0 years. Children with IBS and lactose maldigestion had more frequent abdominal pain than children without these conditions, but they required less medication for relief of symptoms. CONCLUSIONS. Lactose maldigestion may be a contributory factor in children with IBS, and lactose avoidance in these patients may reduce medication use to relieve symptoms. PMID- 10456716 TI - Quality of care for Medicare patients hospitalized with heart failure in rural Georgia. AB - BACKGROUND: It is not known whether quality of care for congestive heart failure (CHF) at rural hospitals is similar to that in larger, urban hospitals. METHODS: We reviewed hospital charts for 310 Medicare patients hospitalized with CHF at six hospitals in rural Georgia. RESULTS: Of the 310 patients, 101 (33%) had left ventricular systolic dysfunction, and 60 (19%) had preserved systolic function. Information on left ventricular function was not available for 48% (range, 29% to 87% across the six hospitals). Among patients with systolic dysfunction, 77% were prescribed an angiotensin converting enzyme (ACE) inhibitor at discharge, and 73% were prescribed digoxin. However, the mean daily ACE inhibitor dose was only 48% of the recommended target dose. Only 30% of all patients with atrial fibrillation were prescribed warfarin. CONCLUSIONS: Overall quality of care for CHF at rural hospitals appears similar to that in other settings, though many patients may not receive evaluation of ventricular function. PMID- 10456717 TI - Graduate surgical trainee attitudes toward postoperative thromboprophylaxis. AB - BACKGROUND: Postoperative deep vein thrombosis (DVT) results in significant morbidity and mortality. Appropriate patient risk assessment and proper use of prophylactic measures is crucial. METHODS: We conducted a confidential survey of general surgery house staff. The survey addressed DVT risk factor recognition, proximal and distal DVT absolute risk estimation, DVT prophylaxis strategies, and vena cava filter and low-molecular-weight heparin (LMWH) use. RESULTS: Obesity, immobility, malignancy, and previous DVT were overwhelmingly recognized as risk factors. Age >40, recent myocardial infarction, lupus anticoagulant, varicose veins, and factor V Leiden were inadequately recognized as risks. Deep vein thrombosis risk in the setting of cancer was underestimated. Most selected appropriate prophylactic strategies; many misunderstood vena cava filter and LMWH indications. CONCLUSIONS: We have identified specific areas of misunderstanding about DVT risk and prophylaxis that, if appropriately addressed during educational sessions, will enable young surgeons to make safer and more effective future decisions regarding thromboprophylaxis. PMID- 10456718 TI - Folk remedy use in the inner city. AB - BACKGROUND: Folk remedies (FRs) are common in some minority communities within the United States. However, little information is available about the use of FRs in the inner city. The objectives of this study were to identify the prevalence and types of FRs used in the inner city, to characterize the population using FRs, and to study patient attitudes toward discussing FRs with physicians. METHODS: We interviewed 71 patients over the age of 18 who visited an inner city ambulatory clinic. RESULTS: The rate of FR use ranged from 10% to 50% among common medical conditions, with most patients (59%) using at least one remedy. Folk remedy use did not correlate with patient age. Most patients used FRs because they believed them to be efficacious, and most felt comfortable discussing FRs with their physicians. CONCLUSIONS: Inner city residents of all ages use FRs to treat illness. These FRs are benign and consist of common household items. Health care workers should feel comfortable discussing FRs with their patients since their patients are comfortable with the topic. PMID- 10456719 TI - Clinical presentation and treatment of a Salmonella bredeney epidemic in Shelby County, Alabama. AB - BACKGROUND: Numerous residents of Shelby County, Alabama, were infected with Salmonella when a restaurant unknowingly served food tainted with the bacterium. Because of the similarity in symptoms caused by other gastrointestinal pathogens and the variability in time of presentation, an outbreak such as this could be confused with one of another pathogenic origin. The pathogen identified, Salmonella bredeney, is a particularly rare cause of food poisoning. It makes up only 0.1% of the Salmonella isolates identified by the Centers for Disease Control and Prevention (CDC) each year. METHODS: We analyzed patient presentations through chart review and combined this information with that obtained from the state laboratories in Montgomery and the Shelby County Health Department. RESULTS: Symptoms were mostly gastrointestinal and ranged greatly in severity. The total number of patients affected in this incident exceeded 170, making it the largest epidemic of its kind in the recent history of Alabama. CONCLUSIONS: The outbreak in Shelby County was caused by an exceedingly rare species of Salmonella. At this time, it is the only outbreak of S bredeney reported in MEDLINE-accessible literature since 1983. PMID- 10456720 TI - Low amniotic fluid volume is poorly identified in singleton and twin pregnancies using the 2 x 2 cm pocket technique of the biophysical profile. AB - BACKGROUND: This study was done to determine the accuracy of the 2 x 2 cm pocket identifying low amniotic fluid (AF) volume in singleton and twin pregnancies. METHODS: The AF volume was evaluated by ultrasonography for the presence of a 2 x 2 cm pocket before amniocentesis. The actual AF volume was then determined by a diazo-dye reaction with subsequent spectrophotometric analysis using paraminohippurate. RESULTS: The AF volume was low in 21 of the 79 singleton pregnancies and normal in 47; hydramnios was present in 11. Among amniotic sacs of the 60 twin pairs, oligohydramnios was found in 33 amniotic sacs. normal AF volume in 80 sacs, and high volume in 7. An AF pocket smaller than 2 x 2 cm was identified in only 3 of the 79 singleton pregnancies and in only 2 of the 120 twin amniotic sacs. CONCLUSIONS: Judging AF volume on the basis of a 2 x 2 cm pocket misses more than 90% of cases of oligohydramnios in singletons and twins. PMID- 10456721 TI - Sporadic Creutzfeldt-Jakob disease presenting as major depression. AB - We describe a 68-year-old white woman who initially had symptoms of major depression and was admitted to a psychiatric hospital where she had electroconvulsive therapy. With failure of psychiatric treatment and subsequent rapidly progressive dementia, she had left frontal brain biopsy. The biopsy revealed spongiform changes, the hallmark of Creutzfeldt-Jakob disease (CJD). We report a case of sporadic CJD with unusual initial presentation of psychiatric symptoms. PMID- 10456722 TI - Transient lumbosacral polyradiculopathy after prostatectomy: association with spinal stenosis. AB - Mononeuropathies are common after pelvic surgery. They are usually the result of unnatural positioning during surgery or faulty restraining devices. Polyneuropathy in the postoperative setting is rare. We report two cases of polyradiculopathy after radical prostatectomy using two different patient positions. Both patients complained of paresthesias and weakness in their lower extremities on postoperative day 1. Neurologic examination in each case was consistent with a polyradiculopathy. Significant spinal stenosis of the lumbosacral spine was found in both patients by magnetic resonance imaging. We propose that spinal stenosis is a risk factor for this type of neurologic injury. PMID- 10456723 TI - Destructive native valve endocarditis caused by Staphylococcus lugdunensis. AB - Coagulase-negative staphylococci are uncommon causes of native valve endocarditis, and the clinical course after valvular infection with these organisms is variable. In clinical practice, species identification is frequently not done, and possible differences in the pathogenicity of various species may be unrecognized. We report a case of Staphylococcus lugdunensis native valve endocarditis associated with valve leaflet perforation and cerebral embolization. This recently described species appears to be more virulent when infecting native cardiac valves than other species of coagulase-negative staphylococci. We review S lugdunensis native valve endocarditis. PMID- 10456724 TI - Rothia endocarditis in a child. AB - Rothia dentocariosa, an organism inhabiting the oral flora, has been reported to cause endocarditis in adults but not in children. Most patients reported with Rothia endocarditis have a history of poor dentition. We report a case of Rothia endocarditis in a child with congenital heart disease without other known risk factors. PMID- 10456725 TI - Correlation of pseudohypoxemia and leukocytosis in chronic lymphocytic leukemia. AB - Pseudohypoxemia has been reported in leukemic patients with extreme leukocytosis, and it is characterized by a low oxygen saturation on arterial blood gas analysis despite normal saturation on pulse oximetry. We report the case of a 51-year-old man with chronic lymphocytic leukemia and an elevated white blood cell (WBC) count after splenectomy, his progressive postoperative pseudohypoxemia gradually improved as the leukocytosis was lowered by chemotherapy. We believe this is the first report to show a statistically significant correlation between the WBC count and the degree of pseudohypoxemia in a patient with leukemia. PMID- 10456726 TI - Whipple's disease presenting as sarcoidosis and valvular heart disease. AB - Whipple's disease is a rare systemic disorder that can present in a variety of ways and is often difficult to diagnose. It can involve almost any organ system and can mimic other diseases both in its symptoms and pathologic appearance. We present a case initially diagnosed as sarcoidosis that was found to be Whipple's disease after pathologic examination of the patient's mitral and aortic valves. PMID- 10456727 TI - Thromboembolism in the right side of the heart. AB - Right-sided cardiac thromboemboli, or pulmonary emboli-in-transit, represent an unusual disease process with high morbidity and mortality. We present a detailed case report and a synopsis of our experience at the University of Mississippi Medical Center between 1990 and 1997 and review the current medical literature. These thrombi and emboli may largely be subdivided into type A--a mobile, serpiginous clot that is probably a mobilized deep vein thrombus--and type B--a rather immobile clot morphologically similar to left-sided heart thrombi that may represent intracardiac thrombosis. The mainstays of therapy include surgical embolectomy and thrombolysis, but there is no clear benefit of one over the other. Treatment should be individualized according to the clot's size and morphology, likelihood of preexisting pulmonary embolism, the patient's cardiopulmonary reserve, comorbid conditions, and local expertise with treatment modalities. PMID- 10456728 TI - Infection of a knee prosthesis with Tsukamurella species. AB - A 69-year-old woman with a history of multiple infections of a postoperative wound from a knee replacement was diagnosed with an infection with Tsukamurella sp. The infection was treated with a course of vancomycin and pipercillin/tazobactam, followed by a course of clarithromycin, ciprofloxacin, and ethambutol. The patient responded well. This represents the first report of a Tsukamurella infection of an artificial joint. PMID- 10456729 TI - Always remember chylothorax. AB - Chylothorax is a rare cause of pleural effusion in association with cardiovascular surgery. A 68-year-old man had a coronary artery bypass graft; 10 days after surgery, he had progressive shortness of breath and a massive left pleural effusion. After chylothorax was diagnosed, it was successfully treated with a thoracostomy tube and total parenteral nutrition. The patient had an uneventful recovery. Nutrition is the most important issue in the treatment of chylothorax. Once chylothorax is identified, nutritional support is the priority, since it will have an important role in the recovery of the patient. Besides our case, we hereby present a short review of the literature regarding the diagnosis and management of this rare entity. PMID- 10456730 TI - Folate deficiency. PMID- 10456731 TI - Obesity management. PMID- 10456732 TI - Hyponatremia associated with paroxetine use. PMID- 10456733 TI - Nucleic acid persistence in heat-killed Escherichia coli O157:H7 from contaminated skim milk. AB - Polymerase chain reaction (PCR) and reverse transcriptase (RT)-PCR using primers targeting 16S rRNA sequences in Escherichia coli O157:H7 were applied to monitor the stability of rDNA and rRNA in cells killed by mild heat treatment (60 degrees C) in skim milk. Serial dilutions of purified RNA and DNA from E. coli O157:H7 in skim milk were amplified by RT-PCR or PCR, respectively, before heat treatment and at time points 0, 6, 12, 24, and 48 h after heating. In general, DNA-PCR provided stronger amplification signals compared to RT-PCR at the corresponding time points with the same PCR primer set, indicating a lower efficiency of RNA amplification compared to that of DNA. Ribosomal RNA and rDNA could be amplified by RT-PCR or PCR from both viable and dead cells throughout the 48-h posttreatment holding period. For RT-PCR, amplification signals decreased in intensity with increased holding time, while the efficiency of amplification of DNA sequences from dead cells remained fairly stable throughout the study. DNA persistence was greater than that of rRNA following cell death by mild heat treatment in skim milk. Skim milk did not appear to accelerate nucleic acid degradation. While rRNA was less stable than DNA, its detection by RT-PCR may not be appropriate as an exclusive indicator of cell viability in minimally processed foods. PMID- 10456734 TI - Survival of enterohemorrhagic Escherichia coli O157:H7 in bovine feces applied to lettuce and the effectiveness of chlorinated water as a disinfectant. AB - Bovine feces are a potential vehicle for transmitting enterohemorrhagic Escherichia coli O157:H7 to humans. A study was undertaken to determine survival characteristics of E. coli O157:H7 on iceberg lettuce using 0.1% peptone water and bovine feces as carriers for inocula. Four levels of inoculum, ranging from 10(0) to 10(5) CFU of E. coli O157:H7 per g of lettuce, were applied. Populations surviving on lettuce stored at 4 degrees C were monitored for up to 15 days. Regardless of the type of carrier, viable cells of E. coli O157:H7 were detected on lettuce after 15 days, even when the initial inoculum was 10(0) to 10(1) CFU/g. Spray treatments of lettuce with 200 ppm chlorine solution or deionized water were equally effective in killing or removing E. coli O157:H7 from lettuce. Holding lettuce for 5 min after spray treatment was not more effective in reducing populations than holding for 1 min before rinsing with water. Prevention of contamination of lettuce with bovine feces that may harbor E. coli O157:H7 as well as other infectious microorganisms is essential to minimizing the risk of illness. The development of sanitizers more efficacious than chlorine for the removal of pathogens from raw fruits and vegetable is needed. PMID- 10456735 TI - Behavior of enterohemorrhagic Escherichia coli O157:H7 on alfalfa sprouts during the sprouting process as influenced by treatments with various chemicals. AB - The behavior of Escherichia coli O157:H7 on alfalfa seeds subjected to conditions similar to those used commercially to grow and market sprouts as it is affected by applications of NaOCl, Ca(OCl)2, acidified NaClO2, acidified ClO2, Na3PO4, Vegi-Clean, Tsunami, Vortexx, or H2O2 at various stages of the sprouting process was determined. Application of 2,000 ppm of NaOCl, 200 and 2,000 ppm of Ca(OCl)2, 500 ppm of acidified ClO2, 10,000 ppm of Vegi-Clean, 80 ppm of Tsunami, or 40 and 80 ppm of Vortexx to germinated seeds significantly reduced the population of E. coli O157:H7. With the exception of acidified NaOCl2 at 1,200 ppm, spray applications of these chemicals did not significantly reduce populations or control the growth of E. coli O157:H7 on alfalfa sprouts during the sprouting process. Populations of E. coli on alfalfa sprouts peaked at 6 to 7 log10 CFU/g 48 h after initiation of the sprouting process and remained stable despite further spraying with chemicals. The population of E. coli O157:H7 on sprouts as they entered cold storage at 9 +/- 2 degrees C remained essentially unchanged for up to 6 days. None of the chemical treatments evaluated was able to eliminate or satisfactorily reduce E. coli O157:H7 on alfalfa seeds and sprouts. Observations on the ability of E. coli O157:H7 to grow during production of alfalfa sprouts not subjected to chemical treatments are similar to those from a previous study in our laboratory on the behavior of Salmonella Stanley. Our results do not reveal a chemical treatment method to eliminate the pathogen from alfalfa sprouts. We have demonstrated that currently recommended procedures for sanitizing alfalfa seeds fail to eliminate E. coli O157:H7 and that the pathogen can grow to populations exceeding 7 1og10 CFU/g of sprouts produced using techniques not dissimilar to those used in the sprout industry. PMID- 10456736 TI - Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes on plastic kitchen cutting boards by electrolyzed oxidizing water. AB - One milliliter of culture containing a five-strain mixture of Escherichia coli O157:H7 (approximately 10(10) CFU) was inoculated on a 100-cm2 area marked on unscarred cutting boards. Following inoculation, the boards were air-dried under a laminar flow hood for 1 h, immersed in 2 liters of electrolyzed oxidizing water or sterile deionized water at 23 degrees C or 35 degrees C for 10 or 20 min; 45 degrees C for 5 or 10 min; or 55 degrees C for 5 min. After each temperature-time combination, the surviving population of the pathogen on cutting boards and in soaking water was determined. Soaking of inoculated cutting boards in electrolyzed oxidizing water reduced E. coli O157:H7 populations by > or = 5.0 log CFU/100 cm2 on cutting boards. However, immersion of cutting boards in deionized water decreased the pathogen count only by 1.0 to 1.5 log CFU/100 cm2. Treatment of cutting boards inoculated with Listeria monocytogenes in electrolyzed oxidizing water at selected temperature-time combinations (23 degrees C for 20 min, 35 degrees C for 10 min, and 45 degrees C for 10 min) substantially reduced the populations of L. monocytogenes in comparison to the counts recovered from the boards immersed in deionized water. E. coli O157:H7 and L. monocytogenes were not detected in electrolyzed oxidizing water after soaking treatment, whereas the pathogens survived in the deionized water used for soaking the cutting boards. This study revealed that immersion of kitchen cutting boards in electrolyzed oxidizing water could be used as an effective method for inactivating foodborne pathogens on smooth, plastic cutting boards. PMID- 10456737 TI - Influence of guar gum on the thermal stability of Listeria innocua, Listeria monocytogenes, and gamma-glutamyl transpeptidase during high-temperature short time pasteurization of bovine milk. AB - Mathematical models describing the thermal inactivation of gamma-glutamyl transpeptidase (TP) and Listeria innocua in milk during high-temperature short time (HTST) pasteurization were validated with data from TP, L. innocua, and L. monocytogenes trials in guar gum-milk. Holding tube times were determined for turbulent flow using water, and for laminar flow using a guar gum (0.27% wt/wt) sucrose (5.3% wt/wt)-water mixture. Inactivation of TP and L. innocua was lower in a solution of guar gum (0.25% wt/wt) in whole milk than was predicted by models derived from studies with whole milk alone. Use of laminar flow timings improved model fit but did not completely account for the observed protective effect. L. monocytogenes survival was close to that predicted by the L. innocua model, although some protection was afforded this pathogen under laminar flow. Considerable intertrial variability was noted for L. monocytogenes. Risk analysis simulations using @RISK, a Lotus 1-2-3W add-in, were used to account for intertrial variability. Simulated log10 %reductions consistently underpredicted experimental L. monocytogenes survival (fail-safe), thus the L. innocua model derived in milk is suitable for estimating L. monocytogenes survival in viscous products. Increased thermal tolerance during laminar flow may be attributed to the protective effect of stabilizer. PMID- 10456738 TI - Clostridium botulinum spores and toxin in mascarpone cheese and other milk products. AB - A total of 1,017 mascarpone cheese samples, collected at retail, were analyzed for Clostridium botulinum spores and toxin, aerobic mesophilic spore counts, as well as pH, a(w) (water activity), and Eh (oxidation-reduction potential). In addition 260 samples from other dairy products were also analyzed for spores and botulinum toxin. Experiments were carried out on naturally and artificially contaminated mascarpone to investigate the influence of different temperature conditions on toxin production by C. botulinum. Three hundred and thirty-one samples (32.5%) of mascarpone were positive for botulinal spores, and 7 (0.8%) of the 878 samples produced at the plant involved in an outbreak of foodborne botulism also contained toxin type A. The chemical-physical parameters (pH, a(w), Eh) of all samples were compatible with C. botulinum growth and toxinogenesis. Of the other milk products, 2.7% were positive for C. botulinum spores. Growth and toxin formation occurred in naturally and experimentally contaminated mascarpone samples after 3 and 4 days of incubation at 28 degrees C, respectively. PMID- 10456739 TI - Toxin production by Clostridium botulinum in pasteurized milk treated with carbon dioxide. AB - The addition of carbon dioxide to milk at levels of <20 mM inhibits the growth of selected spoilage organisms and extends refrigerated shelf life. Our objective was to determine if the addition of CO2 influenced the risk of botulism from milk. Carbon dioxide was added to pasteurized 2% fat milk at approximately 0, 9.1, or 18.2 mM using a commercial gas-injection system. The milk was inoculated with a 10-strain mixture of proteolytic and nonproteolytic Clostridium botulinum spore strains to yield 10(1) to 10(2) spores/ml. Milk was stored at 6.1 or 21 degrees C for 60 or 6 days, respectively, in sealed glass jars or high-density polyethylene plastic bottles. Milk stored at 21 degrees C curdled and exhibited a yogurt-like odor at 2 days and was putrid at 4 days. Botulinal toxin was detected in 9.1 mM CO2 milk at 4 days and in all treatments after 6 days of storage at 21 degrees C. All toxic samples were grossly spoiled based on sensory evaluation at the time toxin was detected. Although botulinal toxin appeared earlier in milk treated with 9.1 mM CO2 compared to both the 18.2 mM and untreated milk, gross spoilage would act as a deterrent to consumption of toxic milk. No botulinal toxin was detected in any treatment stored at 6.1 degrees C for 60 days. At 6.1 degrees C, the standard plate counts (SPCs) were generally lower in the CO2 treated samples than in controls, with 18.2 mM CO2 milk having the lowest SPC. These data indicate that the low-level addition of CO2 retards spoilage of pasteurized milk at refrigeration temperatures and does not increase the risk of botulism from treated milk stored at refrigeration or abuse temperatures. PMID- 10456740 TI - The microbiological quality of cooked rice from restaurants and take-away premises in the United Kingdom. AB - The microbiological quality of 4,162 samples of cooked rice from restaurants and take-away premises in the United Kingdom was examined, including ready-to-eat rice purchased at point-of-sale and rice that was stored precooked for reheating on demand. The majority of point-of-sale cooked rice samples (1,855 of 1,972; 94%) were of acceptable microbiological quality, but 15 (1%) samples were of unacceptable quality (Bacillus spp. and B. cereus, > or = 10(5) CFU/g; Escherichia coli, > or = 10(4) CFU/g), indicating a potential risk to health. The prevalence of Bacillus spp., B. cereus, and E. coli was significantly greater in precooked stored rice than in point-of-sale cooked rice (P < 0.005 to 0.0005). Bacillus spp. (> or = 10(4) CFU/g), B. cereus (> or = 10(4) CFU/g), and E. coli (> or = 10(2) CFU/g) were present in 7%, 2%, and 9% of precooked stored samples, respectively, compared to 2%, 0.5%, and 1%, respectively in point-of-sale samples. Although final heating at the point of sale reduces the levels of microorganisms present in rice it will not inactivate the B. cereus emetic toxin if present. Rice from Indian premises was of poorer microbiological quality than that from Chinese and other premises. Although most point-of-sale cooked rice samples (94%) were of an acceptable microbiological quality, evidence from this study indicates that the microbiological quality of cooked rice sold from certain outlets in the UK is of concern. PMID- 10456741 TI - Alicyclobacillus in orange juice: occurrence and heat resistance of spores. AB - Spore suspensions of a pure culture of Alicyclobacillus acidoterrestris DSM 2498 were submitted to different heat treatments (60 degrees C for 60 min, 60 degrees C for 30 min, 70 degrees C for 20 min, 80 degrees C for 5 min, 80 degrees C for 10 min, 80 degrees C for 30 min, and boiling for 5 min) to determine the best activation conditions in orange juice. The best treatment for spore activation was shown to be 70 degrees C/20 min. Seventy-five samples of concentrated orange juice from 11 different suppliers were examined for the presence of thermophilic acid-tolerant spore formers by the most probable number technique using Bacillus acidocaldarius medium (BAM broth) and incubation at 44 degrees C for 5 days after a prior spore activation. After incubation, isolation was carried out using BAM agar medium incubating at 44 degrees C for 5 days. Typical colonies were submitted to a microscopic examination, evaluation for the presence of spores, and various biochemical tests. Of the orange juice samples examined, 14.7% were found to be positive for Alicyclobacillus. The thermal death time open tube method was used to determine the heat resistance of the spores of strains confirmed as being Alicyclobacillus. The D-values determined were in the range from 60.8 to 94.5 min at 85 degrees C, 10.0 to 20.6 min at 90 degrees C, and 2.5 to 8.7 min at 95 degrees C. The z-values were between 7.2 degrees C and 11.3 degrees C. The results demonstrated the occurrence of Alicyclobacillus in orange juice and the high heat resistance of the spores that could survive the heat treatments normally applied in the processing of orange juice. PMID- 10456742 TI - Verifying apple cider plant sanitation and hazard analysis critical control point programs: choice of indicator bacteria and testing methods. AB - The objectives of this study were (i) to evaluate the survival of coliforms, Escherichia coli, and enterococci in refrigerated apple cider; (ii) to develop simple and inexpensive presumptive methods for detection of these bacteria; (iii) to perform a field survey to determine the prevalence of these bacteria on apples and in apple cider; and (iv) based on our results, to recommend the most useful of these three indicator groups for use in verifying apple cider processing plant sanitation and hazard analysis critical control point (HACCP) programs. Eight of 10 coliform strains (5 E. coli, 1 Enterobacter aerogenes, and 2 Klebsiella spp.) inoculated into preservative-free apple cider (pH 3.4, 13.3(o) Brix) survived well at 4 degrees C for 6 days (< or = 3.0 log10 CFU/ml decrease). Of 21 enterococci strains (Enterococcus faecalis, E. faecium, and E. durans), only 2 E. durans and 3 E. faecium strains survived well. Simple broth-based colorimetric methods were developed that detected the presence of approximately 10 cells of coliforms or enterococci. In three field studies, samples of unwashed apples (drops and picked), washed apples, and freshly pressed cider were presumptively analyzed for total coliforms, E. coli, and enterococci using qualitative and/or quantitative methods. Drop apples were more likely than picked apples to be contaminated with E. coli (26.7% vs. 0%) and enterococci (20% vs. 0%). Washing had little effect on coliform populations and in one field study was associated with increased numbers. Total coliform populations in cider ranged from < 1 CFU/ml to > 738 most probable number/ml, depending on the enumeration method used and the sample origin. E. coli was not recovered from washed apples or cider, but enterococci were present on 13% of washed apple samples. The qualitative coliform method successfully detected these bacteria on apples and in cider. Based on its exclusively fecal origin, good survival in apple cider, and association with drop apples, we conclude that E. coli is the most useful organism for verifying apple cider sanitation and HACCP programs. PMID- 10456743 TI - Inactivation of Lactobacillus helveticus bacteriophages by thermal and chemical treatments. AB - The effect of several biocides and thermal treatments on the viability of four Lactobacillus helveticus phages was investigated. Times to achieve 99% inactivation of phages at 63 degrees C and 72 degrees C in three suspension media were calculated. The three suspension media were tris magnesium gelatin buffer (10 mM Tris-HCl, 10 mM MgSO4, and 0.1% wt/vol gelatin), reconstituted skim milk sterile reconstituted commercial nonfat dry skim milk, and Man Rogosa Sharpe broth. The thermal resistance depended on the phage considered, but a treatment of 5 min at 90 degrees C produced a total inactivation of high titer suspensions of all phages studied. The results obtained for the three tested media did not allow us to establish a clear difference among them, since some phages were more heat resistant in Man Rogosa Sharpe broth and others in tris magnesium gelatin buffer. From the investigation on biocides, we established that sodium hypochlorite at a concentration of 100 ppm was very effective in inactivating phages. The suitability of ethanol 75%, commonly used to disinfect utensils and laboratory equipment, was confirmed. Isopropanol turned out to be, in general, less effective than ethanol at the assayed concentrations. In contrast, peracetic acid (0.15%) was found to be an effective biocide for the complete inactivation of all phages studied after 5 min of exposure. The results allowed us to establish a basis for adopting the most effective thermal and chemical treatments for inactivating phages in dairy plant and laboratory environments. PMID- 10456744 TI - Identification and characterization of two bacteriocin-producing bacteria isolated from garlic and ginger root. AB - Two bacteriocin-producing bacterial strains were isolated from garlic and ginger root by the agar overlay method. The bacteria were identified by 16S rRNA sequence analyses and fermentation patterns as Leuconostoc mesenteroides (garlic isolate) and Lactococcus lactis (ginger isolate). The bacteriocins were assigned the names leucocin BC2 and lactocin GI3, respectively. Physiochemical properties and antimicrobial spectra of the bacteriocins were determined by the spot-on-lawn method. Both bacteriocins were inhibited by proteolytic enzymes. Leucocin BC2 exhibited a narrow antimicrobial spectrum, inhibiting only Bacillus, Enterococcus, and Listeria species. Lactocin GI3 had a broader spectrum, inhibiting Bacillus, Clostridium, Listeria, Enterococcus, Leuconostoc, Pediococcus, and Staphylococcus species. Both bacteriocins remained active when heated at 90 degrees C for 15 min or 120 degrees C for 20 min. Leucocin BC2 assayed at 37 degrees C showed an inhibitory activity of 1,600 AU/ml, whereas at 8 degrees C the activity was 12,800 AU/ml. Conversely, lactocin GI3 activity was the same at both assay temperatures. Both bacteriocins remained active over a pH range of 2.0 to 9.0 and in various organic solvents. The activity of leucocin BC2 was increased when treated with 0.5% acetic acid and 0.5% lactic acid, whereas lactocin GI3 activity was decreased with either acid. The molecular mass values were 3.7 kDa for leucocin BC2 and 3.9 kDa for lactocin GI3. These results show that the inhibitory substances produced by the bacteria isolated from garlic and ginger are bacteriocins that appear to be different in some characteristics from previously reported bacteriocins. PMID- 10456745 TI - Screening of bile resistance and bile precipitation in lactic acid bacteria and bifidobacteria. AB - A modification of the ecometric method was developed for a rapid screening of bile resistance in lactic acid bacteria and bifidobacteria. Validation of the MEM bile assay (modified ecometric method) was performed comparing the bile resistance index (RIbile) and the bile resistance ratio (R%). Most Bifidobacterium strains assayed were bile sensitive (83.3%), while only 62.1% of lactobacilli showed that behavior. Some bifidobacterial strains (55.6%) showed a crystalline precipitate when grown on solid medium supplemented with 0.5% ox bile. The crystalline structures produced by B. pseudolongum CIDCA 531 were isolated and analyzed by optical and scanning electron microscopy, thin-layer chromatography, melting point, and specific cholesterol reactions. Those studies confirmed the presence of cholesterol in these crystalline structures. On the other hand, none of the lactobacilli and streptococci studied had the ability to produce crystalline precipitates. PMID- 10456746 TI - Microbial shelf life determination of vacuum-packaged fresh beef treated with polylactic acid, lactic acid, and nisin solutions. AB - The effectiveness of polylactic acid, lactic acid, nisin, and combinations of the acids and nisin on extending the shelf-life of raw beef was determined. Fresh beef pieces (5 by 5 by 2.5 cm) were dipped in a solution of 2% low molecular weight polylactic acid (LMW-PLA), 2% lactic acid (LA), 200 IU of nisin per ml, or the combinations of nisin in either 2% LMW-PLA or 2% LA. The samples were then drip-dried, vacuum-packaged, and stored at 4 degrees C for up to 56 days. The beef surface pH values and numbers of psychrotrophic aerobic bacteria, psychrotrophic and mesophilic Enterobacteriaceae, Pseudomonas, and Lactobacillus were determined weekly for 56 days. The average surface pH values of the beef samples treated with 2% LMW-PLA or the combination of 200 IU of nisin per ml and 2% LMW-PLA were significantly reduced to 5.19 and 5.17, respectively, at day 0 (P < or = 0.05), while those decontaminated with 2% LA or 200 IU of nisin per ml in 2% LA solution were significantly decreased from 5.62 to 4.98 and 4.96, respectively. The 2% LMW-PLA, 2% LA, or the combinations of each acid and nisin showed immediate inhibitory effects on psychrotrophic aerobic bacteria (1.94, 2.36, 2.59, and 1.76 log reduction, respectively), psychrotrophic Enterobacteriaceae (1.37, 1.86, 1.77, and 1.35 log reduction, respectively), mesophilic Enterobacteriaceae (1.00, 1.00, 0.82, and 0.68 log reduction, respectively), and Pseudomonas (1.77, 1.57, 1.76, and 1.41 log reduction, respectively) on fresh beef (P < or = 0.05). The reduction was evident up to 56 days as seen by the numbers of Enterobacteriaceae and Pseudomonas (P < or = 0.05). Because there was no interaction between treatments and storage times, the data in each period were combined and presented as effect of treatments on overall microbial counts of fresh beef. It was found that 2% LMW-PLA, 2% LA, and the combinations of each acid and nisin significantly lowered the population of the above organisms compared with the untreated control, water, or nisin alone (P < or = 0.05). PMID- 10456747 TI - A National Australian Food Safety Telephone Survey. AB - A sample of 1,203 randomly selected Australian households participated in a national telephone food safety survey. All respondents were aged 18 years or over, were the main grocery buyers of the household, purchased red meat products at least once a month, and regularly prepared food in the household. There were significant gaps observed in the food safety knowledge of many respondents surveyed. Forty percent of respondents thawed raw meat at room temperature, 85% allowed cooked foods to cool at room temperature before refrigerating, and almost 70% of respondents were not aware of the correct refrigeration temperature for storage of perishable food. Almost 25% of respondents failed to identify that washing hands before handling food and during food preparation was important in reducing the risk of cross-contamination and possible foodborne illness. Seventy five percent of the respondents recognized that there was a likelihood of foodborne illness occurring in the home, and 25% of respondents had changed their eating habits because of publicity surrounding food poisoning outbreaks. The findings raise important concerns about domestic food handling practices in Australian homes and the level of food safety knowledge in the community generally. PMID- 10456748 TI - Antimicrobial and antioxidant compounds in the nonvolatile fraction of expressed orange essential oil. AB - Three permethoxylated flavones, dehydroabietic acid and linoleil monoglyceride were isolated from the nonvolatile residue of orange essential oil. All of them exhibited antifungal activity against phytopathogenic species and food contaminants. The isolated hexa- and heptamethoxy flavones exhibited important fungicidal activity against Geotrichum candidum, which is not inhibited by the commercial broadspectrum fungicide Benomyl (methyl-1-[butylcarbamoyl]-2 benzimidazolecarbamate); in addition, these compounds also exhibited effective antioxidant activities, similar to that shown by tert-butyl-4-hydroxyanisol, one of the most commonly used synthetic antioxidants; alpha-tocopherol, the most widely used natural antioxidant, was also isolated. PMID- 10456749 TI - Histamine and cadaverine production by bacteria isolated from fresh and frozen albacore (Thunnus alalunga). AB - Two hundred twenty-seven bacterial strains were isolated from fresh and frozen albacore stored either at -18 or -25 degrees C and investigated for their abilities to produce biogenic amines. As a preliminary screening, all 227 strains were tested in either Niven or Niven modified medium, which allowed the selection of 25 presumptive histamine-producing strains. High-pressure liquid chromatography revealed that only 10 of the 25 strains selected were able to produce low histamine concentrations (<25 ppm) in tryptic soy broth medium supplemented with 2% histidine. None of the 25 strains tested produced putrescine or spermine, whereas 6 strains produced spermidine. Histamine production by Stenotrophomonas maltophilia strain 25MC6 was not prevented at 4 degrees C, and the levels of this amine reached concentrations of 25.8 ppm after 6 days. Three S. maltophilia strains showed strong lysine-decarboxylating activity. Their cadaverine formation capacity was determined by high-pressure liquid chromatography in tryptic soy broth supplemented with 1% lysine; this revealed that the three S. maltophilia strains tested produced more than 700 ppm of cadaverine during the first 24 h of incubation at 37 degrees C. S. maltophilia strain 15MF, initially obtained from fresh albacore tuna, produced up to 2,399 ppm and 4,820 ppm of cadaverine after 24 and 48 h of incubation at 37 degrees C, respectively. To our knowledge, this is the first report on histamine and cadaverine production by strains of the species S. maltophilia, previously known as Pseudomonas and Xanthomonas maltophilia, isolated from fresh and frozen albacore tuna. PMID- 10456750 TI - Inhibition of food-related pathogenic bacteria by god-transformed Penicillium nalgiovense strains. AB - Two strains of Penicillium nalgiovense, which carried the god gene of Aspergillus niger and had increased glucose oxidase (GOD) activity compared with the wild type strain, were tested for their ability to suppress the growth of certain food related pathogenic bacteria. In contrast to the wild type, which showed no antibacterial effect when grown in mixed culture with different bacteria, the two transformed strains were highly antagonistic. The strain that expressed higher amounts of GOD in general had higher inhibitory activity. Both strains showed antibacterial activity against Listeria monocytogenes, Salmonella Enteritidis, and Staphylococcus aureus. The inhibitory activity was dependent on the glucose concentration in the medium. S. aureus was completely inhibited at 1% glucose in the presence of the higher GOD-producing transformant. In contrast, if arabinose was used as a carbon source, no inhibition occurred. If catalase was added to the medium, the inhibitory activity of the transformants was completely inactivated, indicating that the hydrogen peroxide produced was responsible for the antibacterial activity of the transformants. PMID- 10456751 TI - Recovery of Escherichia coli Biotype I and Enterococcus spp. during refrigerated storage of beef carcasses inoculated with a fecal slurry. AB - Three beef front quarters/carcasses were inoculated with a slurry of cattle manure. During storage at 4 degrees C, two sponge samples from each of three sites (i.e., 100 cm2 from each of two fat surfaces and 100 cm2 from a lean surface) were taken from each of the three carcasses on days 0, 1, 3, 7, and 10 after inoculation. The initial numbers of Escherichia coli averaged 2.0 log10 CFU/cm2 (1.21 to 2.47 log10 CFU/cm2) using the Petrifilm method and 2.09 log10 most probable number (MPN)/cm2 (0.88 to 2.96 log10 MPN/cm2) using the MPN method. The initial numbers of enterococci averaged 3.34 log10 CFU/cm2 (3.07 to 3.79 log10 CFU/cm2) using kanamycin esculin azide agar. In general, an appreciable reduction in the numbers of E. coli occurred during the first 24 h of storage; for the Petrifilm method an average reduction of 1.37 log10 CFU/cm2 (0.69 to 1.71 log10 CFU/cm2) was observed, and for the MPN method an average reduction of 1.52 log10 MPN/cm2 (0.47 to 2.08 log10 MPN/cm2) was observed. E. coli were not detected (<-0.12 log10 CFU/cm2) using Petrifilm on day 7 of the storage period on two (initial counts of 1.21 and 2.29 log10 CFU/cm2) of the three carcasses. However, viable E. coli cells were recovered from these two carcasses after a 24 h enrichment at 37 degrees C in EC broth. Viable E. coli cells were detected at levels of -0.10 log10 CFU/cm2 on the third carcass (initial count of 2.47 log10 CFU/cm2) after 7 days at 4 degrees C. No significant difference in recovery of viable cells was observed between the MPN and Petrifilm methods on days 0, 1, and 3 (P > 0.05). However, viable E. coli cells were recovered from all three carcasses by the MPN method on day 7 at an average of -0.29 log10 MPN/ cm2 (-0.6 to -0.1 log10 MPN/cm2). On day 10, viable cells were recovered by the MPN method from two of the three carcasses at -0.63 and -0.48 log10 MPN/cm2 but were not recovered from the remaining carcass (<-0.8 log10 MPN/cm2). Similar to E. coli, the greatest reduction (average of 1.26 log10 CFU/cm2, range = 1.06 to 1.45 log10 CFU/cm2) in the numbers of enterococci occurred during the first 24 h of storage. Because of higher initial numbers and a slightly slower rate of decrease, the numbers of Enterococcus spp. were significantly higher (P < 0.017) than the numbers of E. coli Biotype I after 3, 7, and 10 days of storage. These results suggest that enterococci may be useful as an indicator of fecal contamination of beef carcasses. PMID- 10456752 TI - Evaluation of botulinal toxin production in packaged fresh-cut cantaloupe and honeydew melons. AB - The ability of Clostridium botulinum to produce toxin on cubed, packaged melons was investigated relative to microbial spoilage at various incubation temperatures and in different packaging systems. Freshly cut cubes (approximately 2.5 cm3) of cantaloupe and honeydew melons were surface inoculated with a 10 strain mixture of proteolytic and nonproteolytic spores of C. botulinum (10 to 15 cubes per package; approximately 100 total spores per package). To initially evaluate toxin production and spoilage in a passively modified atmosphere, melon cubes were loosely packaged in air in polyethylene pouches, sealed, and incubated at 7 or 15 degrees C for up to 21 days. At various sampling intervals, samples were tested for headspace oxygen and carbon dioxide levels, pH, presence of botulinal toxin, aerobic and anaerobic plate counts, and counts of yeasts and molds. During incubation, headspace oxygen levels decreased, headspace carbon dioxide levels increased, aerobic and anaerobic plate counts increased, and the pH remained constant or decreased slightly. Botulinal toxin was not detected in any cantaloupe samples or in honeydew samples incubated at 7 degrees C. Botulinal toxin was detected in some honeydew samples at 15 degrees C after 9 days of incubation, but the toxic honeydews were severely spoiled and considered organoleptically unacceptable. A similar second experiment was performed in which half of the melon cubes were treated with UV light to inactivate vegetative organisms before packaging, and these were incubated at 7, 15, or 27 degrees C. In this second experiment, toxin production occurred in the UV-treated samples at 15 degrees C with gross spoilage and at 27 degrees C with only marginal spoilage. These data indicate that inhibition of spoilage organisms with UV light could result in botulinal toxin formation in packaged melons before overt spoilage. PMID- 10456753 TI - Disinfection of mung bean seed with gaseous acetic acid. AB - Mung bean seed inoculated with Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes (3 to 5 log CFU/g) was exposed to gaseous acetic acid in an aluminum fumigation chamber. Salmonella Typhimurium and E. coli O157:H7 were not detected by enrichment of seeds treated with 242 microl of acetic acid per liter of air for 12 h at 45 degrees C. L. monocytogenes was recovered by enrichment from two of 10 25-g seed samples treated in this manner. Fumigation with gaseous acetic acid was also lethal to indigenous bacteria and fungi on mung bean seed. The treatment did not significantly reduce seed germination rates, and no differences in surface microstructure were observed between treated and untreated seed viewed by scanning electron microscopy. PMID- 10456754 TI - Thermal inactivation kinetics of Bacillus stearothermophilus spores using a linear temperature program. AB - A systematic study of the inactivation kinetics of Bacillus stearothermophilus spores was carried out in nonisothermic heating conditions using a linear temperature increase program and analyzing the experimental data by means of a one-step nonlinear regression. The D and z values estimated are close to those obtained in isothermic conditions and estimated by using a two-step model, first D values are calculated, and then in the second step a z value is deduced (D(121 degrees C) = 3.08 and 4.38 min, respectively, and z = 7 and 7.9 degrees C, respectively). No convergence problems were observed when using the one-step nonlinear regression proposed. The results indicated that the methodology applied in this study can be used to obtain kinetic data for bacterial spores, which could mean a significant reduction in the amount of experimental work employed to generate these data. PMID- 10456755 TI - Stability of deoxynivalenol in heat-treated foods. AB - The effects of high-temperature and -pressure processing of foods spiked with deoxynivalenol (DON) were examined. In extruded corn grits, extruded dry dog food, and autoclaved moist dog food, there were no significant reductions (P < 0.05) in DON after processing. Autoclaved cream-style corn showed a reduction in DON of only 12%. Overall, DON was stabile to the high temperature and pressure processes tested. The use of an alpha-amylase in the extraction method for analysis by an enzyme-linked immunosorbant assay (ELISA) improved the recovery of DON from the spiked extruded and autoclaved products by as much as 26% over the standard ELISA method. PMID- 10456756 TI - Epilepsy in the first year of life: a review. AB - The prevalence of epilepsy is highest during the first year of life. Approximately 50% of patients with seizure onset between 1 and 12 months of age suffer from West syndrome. The rest have one of 12 well-delineated epilepsy syndromes, or in most cases have what are classified as generalized or partial seizures. The outcome of infants with partial seizures is significantly worse than that of patients with generalized seizures. This is true not only for symptomatic partial seizures but also for a subgroup of infants with cryptogenic partial seizures. PMID- 10456757 TI - Risk of excessive weight gain in epileptic children treated with valproate. AB - We sought to identify factors associated with excessive weight gain in children treated with valproate, excluding patients fed by gastrostomy or treated with medications known to affect appetite (eg, stimulants). Weight and height were recorded before treatment and at the time of follow-up; a measure of adiposity, body mass index, was computed and expressed in kg/m2, and weight and height for age were converted to Z-score. Putative risk factors included sex, age at start of treatment, monotherapy at start of treatment, duration of follow-up, mental retardation, seizure type (generalized or partial), etiology (idiopathic or cryptogenic versus remote symptomatic), and dose of valproate. Fifty-five children (30 girls, 25 boys), ranging in age at the start of therapy from 1.8 to 16.9 years were followed for 8.6 to 33.8 months. Forty-three patients had primarily generalized seizures, 34 had idiopathic or cryptogenic epilepsy (including 30 with generalized idiopathic epilepsy), and 18 had mental retardation. Valproate was the first antiepileptic drug for 21 patients, and 43 were on monotherapy at the time of follow-up. Height Z-score decreased significantly in girls but was stable in boys. There was a significant increase in body mass index and in weight Z-score. Body mass index was greater than the 90th percentile for age in 14 patients at the start of treatment and in 20 patients at follow-up. Decrease in height Z-score was significantly correlated with female sex and duration of follow-up. Changes in weight Z-score and body mass index were significantly correlated with initial weight Z-score and initial body mass index, respectively, but not with age at start of therapy, duration of follow-up, sex, seizure type, etiology, dose of valproate, or monotherapy. PMID- 10456758 TI - Crush head injuries in infants and young children neurologic and neuropsychologic sequelae. AB - Neurologic and neuropsychologic sequelae of crush head injury, which is produced by static forces occurring when the head is stationary and pinned against a rigid structure, were studied prospectively in a series of eight children ranging in age from 13 to 32 months. Hospital course, computed tomographic findings, and neurologic and developmental outcomes were examined. All children sustained pronounced cerebral trauma characterized by multiple fractures throughout the calvaria, extra-axial hemorrhages, and parenchymal contusions. Cranial nerve injuries were noted in three and hemiparesis in two of the cases. Two months after the injury, 63% of the children displayed deficits in either IQ or motor functioning. One year after the injury, five of the six children reevaluated had a good recovery. Motor scores were significantly lower than cognitive scores at baseline and showed the greatest degree of improvement over time. Neuropsychologic outcome after brain injury produced by static loading of the head is more favorable than from traumatic brain injury associated with dynamic loading. PMID- 10456759 TI - Modeling analysis of change in neurologic abnormalities in children born prematurely: a novel approach. AB - The purpose of this longitudinal study was to evaluate changes in neurologic abnormalities in 212 preterm and 128 full-term children. Preterm infants with severe medical complications were considered at high risk, while those with milder complications were considered at low risk for neurodevelopmental abnormality. The patterns of change in neurologic status for the high- and low risk preterm and term groups across 6, 12, 24, 40, and 54 months of age were compared using growth modeling analysis. As expected, the term group showed minimal change, while the two preterm groups demonstrated complex, nonlinear patterns of change in neurologic abnormalities. While the results demonstrate improvement over time in both groups of preterm infants, 35% of the children in the high-risk group had neurologic scores in the borderline or abnormal range at 54 months. These findings demonstrate that neurologic deficits are still evident at preschool age in some preterm children and that these deficits are related to the severity of neonatal complications. PMID- 10456760 TI - Restricted unilateral Sydenham's chorea: reversible contralateral striatal hypermetabolism demonstrated on single photon emission computed tomographic scanning. AB - Sydenham's chorea results from group A streptococcus infection and subsequent generation of antineuronal antibodies directed at the caudate nucleus and putamen. Predominantly bilateral, in up to 30% of cases the chorea can be unilaterally restricted. Imaging studies, both structural (magnetic resonance imaging) and functional (positron emission tomography), in patients with bilateral Sydenham's chorea have suggested reversible striatal abnormalities. Two patients with unilateral Sydenham's chorea are presented. Computed tomographic and magnetic resonance imaging were normal in both. However, hexamethylpropylenamine oxime single photon emission tomographic (HMPAO SPECT) studies demonstrated hypermetabolism in the contralateral basal ganglia. Resolution of symptoms in one of the patients coincided with normalization of the SPECT scan. Thus, unilateral striatal hypermetabolism appears to underlie the contralateral chorea observed. A SPECT scan probably should be included in the work-up of new-onset chorea. PMID- 10456761 TI - Plantar response profile of high-risk infants at one year of life. AB - To clarify the plantar reflex profile at 1 year of life in different categories of neurodevelopmental abnormalities, plantar responses were examined prospectively in 204 high-risk infants, of whom 58 developed cerebral palsy, 22 had developmental retardation without motor disturbance, and 124 were normal at a follow-up examination at 3 years of age. The plantar response was extensor in 82.3% of infants subsequently found to be neurologically normal at the first month of life, becoming flexor at the age of 9 and 11 months in 68.5% and 86.3%, respectively. Twenty-one (42.9%) of 49 patients with various types of spastic cerebral palsy demonstrated a combined extensor response (ie, dorsiflexion of the great toe with fanning of the remaining toes) as early as the first month of life. Children with spastic quadriplegia and hemiplegia more frequently demonstrated a combined extensor response compared to diplegic patients. The combined extensor plantar response remains a reliable prognostic clinical tool that contributes to an earlier diagnosis of spastic cerebral palsy as early as the first month of life. PMID- 10456762 TI - Flow cytometric evaluation of defects of the mitochondrial respiratory chain. AB - Cultured human skin fibroblasts from 12 patients with a variety of mitochondrial respiratory chain defects were examined for their capacity to oxidize dihydrorhodamine-123 to the fluorescent molecule rhodamine-123 using a flow cytometer. We found that cells from patients with functional defects in respiratory chain enzymes were less able to oxidize dihydrorhodamine-123 than those of healthy controls. Ten of the cell strains had reduced activity in at least one of the respiratory chain complexes and also showed significantly reduced fluorescence when compared to the mean of eight normal control cell strains. One patient had mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes (with the A3243G mutation) and reduced respiratory chain activities in muscle and liver. Molecular analysis did not show the mutation in cultured skin fibroblasts, and had correspondingly normal fluorescence. The 12th cell strain showed reduced fluorescence but did not reach statistical significance. This strategy could be of use in helping direct further investigations in patients, and in studying the biochemical pathogenesis of mitochondrial DNA mutations in cybrid studies. PMID- 10456763 TI - Rett syndrome: 1H spectroscopic imaging at 4.1 Tesla. AB - Rett syndrome, a neurodevelopmental disorder predominantly affecting girls, is characterized by regression of psychomotor development, communication dysfunction, and hand stereotypies. Brain morphologic studies demonstrate increased neuronal packing density and reduced dendritic arborizations, suggesting an arrest or interruption of normal maturation. Numerous neurotransmitter systems have been implicated. Among these, cerebrospinal fluid glutamate levels are elevated and glutamate receptors, particularly in putamen, are reduced. Therefore, 1H spectroscopy at 4.1 Tesla was used to evaluate glutamate, creatine, and N-acetylaspartate in six girls with Rett syndrome and four normal sibling controls. The ratio of creatine to N-acetylaspartate was significantly elevated in white matter, primarily reflecting reduced N acetylaspartate levels, and normal in gray matter. The glutamate to N acetylaspartate ratio was elevated in gray matter and normal in white matter. These findings are consistent with previous neuropathologic and neurochemical findings and indicate the feasibility of imaging these metabolites in vivo. PMID- 10456764 TI - Folinic acid-responsive neonatal seizures. AB - We report three cases of folinic acid-responsive intractable neonatal seizures. All patients were born at term following normal gestation and delivery. In the first infant, seizures began on the 5th day of life and were unresponsive to phenobarbital, pyridoxine, and valproate, but stopped within 24 hours of initiation of folinic acid treatment at the age of 6 months. Her sibling had died at age 6 months with intractable seizures. In the second infant, seizures began in the 2nd hour of life. These were initially controlled with phenobarbital; however, at 3 months of age she developed status epilepticus refractory to anticonvulsants, steroids, and pyridoxine and she required repeated induction of pentobarbital coma. Seizures stopped within 24 hours of starting folinic acid. Seizures and encephalopathy were noted in the third infant on the 2nd day of life. These were controlled with phenobarbital, but at 8 weeks of age seizures recurred and were difficult to control despite the addition of phenytoin. Immediately after folinic acid was initiated the seizures stopped. Breakthrough seizures in all patients have responded to increases in folinic acid; two of the three remain on standard anticonvulsants. All patients have global developmental delay. Cranial magnetic resonance imaging in the second patient shows diffuse atrophy, and in the third patient shows increased signal on T2 images in the white matter of the frontal and parietal lobes. Analysis of cerebrospinal fluid from these patients using high-performance liquid chromatography with electrochemical detection has consistently revealed an as-yet unidentified compound, which can be used as a marker for this condition. We suggest that cerebrospinal fluid be analyzed for the presence of this compound and a trial of folinic acid be considered in neonates with unexplained early onset intractable seizures. PMID- 10456765 TI - Electroencephalographic findings in ornithine transcarbamylase deficiency. AB - A 3-day-old infant presented with anorexia, irritability, hypotonia, and seizures. Blood ammonia was 2115 micromol/L and amino and organic acid analyses were consistent with ornithine transcarbamylase deficiency. Liver biopsy confirmed only 1% enzyme activity. The patient was treated with hemodialysis. An electroencephalogram (EEG) revealed multifocal independent spike-and-sharp-wave discharges. After initial stabilization he was placed on a low-protein diet with citrulline and phenylbutyrate. Conjugating agents (arginine, sodium benzoate, and sodium phenylacetate) have been added during periods of metabolic decompensation. Although developmentally delayed, the patient has shown signs of clinical improvement and EEG activity has likewise improved with only mild background slowing and no evidence of epileptogenic activity at 4 years of age. A second infant presented at 3 days of age with a similar history, blood ammonia of 1382 micromol/L, and metabolic studies indicative of ornithine transcarbamylase deficiency. EEG showed multifocal independent ictal and interictal discharges. Electrographic abnormalities persisted despite lowering of blood ammonia with hemodialysis and conjugating agents. The patient continued to decline clinically and died on the 7th hospital day. EEG changes parallel the clinical course of ornithine transcarbamylase deficiency and may serve as an objective marker of the effectiveness of therapeutic interventions. PMID- 10456766 TI - How it began and developed: a brief autobiographical sketch. PMID- 10456767 TI - Incidental spina bifida occulta in functional enuresis observed during laser reflexo therapy. AB - The etiology of persistent functional enuresis in children is ill understood. A wide range of therapeutic modalities have been tried. During investigations preceding a trial of laser reflexo therapy in functional enuretic children aged 7 to 10 years spina bifida occulta was observed in 86.6% of subjects. The lesions were predominantly at the L5-6 level. There were no cutaneous stigmata in the form of lipoma, tufts of hair, or dermal sinus at the back. Neurologic examination was unremarkable and there were no associated skeletal malformations. This strong association of spina bifida occulta with functional enuresis is higher than the reported incidence of 16.5% to 34% in normal children. This strong association hitherto unreported, raises an interesting issue in the search for the etiopathogenesis of functional enuresis. PMID- 10456768 TI - Cerebral glucose metabolism in type I alpha-N-acetylgalactosaminidase deficiency: an infantile neuroaxonal dystrophy. AB - Cerebral glucose metabolism was investigated in a 4.8-year-old boy with alpha-N acetylgalactosaminidase deficiency using 2-[18F]fluoro-2-deoxy-D-glucose and positron emission tomography (PET). In comparison to normal values for age, the overall cerebral glucose metabolism was reduced and the regional cerebral glucose metabolism was decreased in proportion to the degree of atrophy. In the supratentorial cortical regions, the hypometabolism was asymmetric. However, the level of regional cerebral glucose metabolism in all cortical regions excluded a persistent vegetative state. In the lentiform nucleus and the head of the caudate, comparatively increased regional cerebral glucose metabolism was documented, similar to findings in neurodegenerative disorders with active epilepsy. In contrast, the infratentorial structures (cerebellar hemispheres, brain stem, mesencephalon, and hypothalamus), which are predominantly affected by the atrophic process, showed distinct and symmetric hypometabolism. Thus, the 2 [18F]-fluoro-2-deoxy-D-glucose PET scans provided additional insight into and correlation of the functional and structural disturbances in type I alpha-N acetylgalactosaminidase deficiency, in addition to documenting the hypometabolism due to brain atrophy. PMID- 10456769 TI - Niaprazine in the treatment of autistic disorder. AB - Niaprazine is a histamine H1-receptor antagonist with marked sedative properties. It has been employed in subjects with behavior and sleep disorders. No data concerning the use of niaprazine in subjects with autistic disorder are reported in the literature. The authors performed an open study to assess niaprazine efficacy in a sample of 25 subjects with autistic disorder and associated behavior and sleep disorders. Niaprazine was administered at 1 mg/kg/day for 60 days. A positive effect was found in 52% of patients, particularly on hyperkinesia, unstable attention, resistance to change and frustration, mild anxiety signs, heteroaggressiveness, and sleep disorders. Statistical comparison between responders and nonresponders showed no influence on niaprazine effect by age over or under 12 years, presence of neurologic signs, epilepsy, or abnormalities seen on brain imaging. Niaprazine was more efficacious in subjects with a mild or moderate degree of mental retardation. No side effects were observed. Because of its sedative effects and good tolerability, niaprazine can be used as a first-choice drug to improve behavior and sleep disorders in patients with autistic disorder. PMID- 10456770 TI - Rett syndrome: photographic evidence of rapid regression. AB - Rett Syndrome is known to occur in females, around the second year, with loss of hand use, onset of stereotypes and acquired microcephaly. Such regression is often very rapid, but this has never been documented. In one of our patients, photographs taken at different times clearly demonstrate the rapid progression of first symptoms. Moreover, in the present case, the occurrence of a febrile illness, which preceded the onset of the neurological picture, support the hypothesis that environmental factors may trigger the onset of Rett Syndrome in genetically predisposed subjects. PMID- 10456771 TI - A physico-chemical study on the polysaccharide ulvan from hot water extraction of the macroalga Ulva. AB - Results of a study on the solution behaviour of the cell-wall polysaccharide named ulvan obtained from hot water extraction of a flour of Ulva 'rigida' are reported. In particular the spectroscopic properties and ion binding capacity of this charged polysaccharide were studied by circular dichroism and isothermal microcalorimetric titrations in order to gain information on the potential exploitation of this low cost biomass. A marked tendency of this polysaccharide to uptake water was evidenced by studying the proton spin-lattice relaxation times of the solvent, T1, embedded in this highly charged polysaccharide. PMID- 10456772 TI - Conformation and association of kappa-carrageenan in the presence of locust bean gum in mixed NaI/CsI solutions from rheology and cryo-TEM. AB - Mixtures of locust bean gum (LBG) with kappa-carrageenan (KC) in 0.1 M aqueous solutions of the mixed salts NaI/CsI were investigated by cryo-transmission electron microscopy (cryo-TEM) and dynamic viscoelastic measurements. Previous studies have shown that as the cesium content is increased in such mixed salt solutions, a transition occurs from molecularly dispersed helices to 'superhelical rods' of KC. We now found that LBG stabilises the superhelical rods, shifting the transition to a lower content of Cs for the mixtures than for KC alone. The formation of superhelical rods was evidenced both by cryo-TEM images and by an onset of thermal hysteresis in the coil helix transition of KC. In the mixtures, the transition temperatures on cooling and heating were insensitive to the proportions of LBG and KC present at all cesium contents. Under conditions where no helix aggregation occurred (no hysteresis) the mixtures showed high tan delta values and low storage moduli. Under aggregated conditions, gels formed, and gels with added LBG had enhanced moduli compared to gels with KC alone. On the basis of these results we propose that LBG associates to the super helical rods of KC. PMID- 10456773 TI - Structure determination and refinement of the Al3+ complex of the D254,256E mutant of Arthrobacter D-xylose isomerase at 2.40 A resolution. Further evidence for inhibitor-induced metal ion movement. AB - The structure of the D254.256E double mutant of Arthrobacter xylose isomerase with Al3+ at both metal-binding sites was determined by the molecular replacement method at a conventional R-factor of 0.179. Binding of the two Al3+ does not alter the overall structure significantly. However, there are local rearrangements in the octahedral co-ordination sphere of the Al3+. The inhibitor molecule moves somewhat away from the active site. Furthermore, evidence was revealed for metal ion movement from site 2(1) to site 2(2) upon double mutation. Xylose isomerase requires two divalent metal cations for activation. The catalytic metal ion is translocated 1.8 A away from its initial position during the catalytic reaction. The fact that both activating and inactivating metals (including Al3+) were found exclusively at a single location in the double mutant was an indication that the consequently missing shuttle may account for the crippled catalytic efficiency. PMID- 10456774 TI - Beta-lactam degradation catalysed by Cd2+ ion in methanol. AB - Kinetic schemes are established for degradation catalysed by Cd2+ ions in methanolic medium for penicillin G, penicillin V and cephalothin, a cephalosporin. Methanolysis of penicillin V and cephalothin occurs with the formation of a single substrate-metal ion intermediate complex, SM, while degradation of penicillin G occurs with the initial formation of two complexes with different stoichiometry, SM and S2M. In each case. degradation is of first order with respect to SM with rate constant values equal to 0.079 min(-1), 0.120 min(-1) and 0.166 min(-1) at 20, 25 and 30 degrees C, respectively, for penicillin G; 0.061 min(-1) at 20 degrees C for penicillin V; and 2.0 x 10(-3) min(-1) at 20 degrees C for cephalothin. Activation energy for the decomposition process of the SM intermediate for penicillin G was calculated to be about 5.5 x 10(4) J/mol. Equilibrium constant values between SM compound and S2M at 20 degrees C (77.1 l/mol), 25 degrees C (45.3 l/mol) and at 30 degrees C (25.7 l/mol) were also calculated as well as the normal enthalpy of this equilibrium. With respect to the reaction products there is evidence that Cd2+ becomes part of their structure, forming complexes between Cd2+ and the product resulting from antibiotic methanolysis (L). Some characteristics of these complexes are discussed. PMID- 10456775 TI - Identification of simian cyclophilin A as a calreticulin-binding protein in yeast two-hybrid screen and demonstration of cyclophilin A interaction with calreticulin. AB - Cyclophilin A (CyPA) was identified as one of the calreticulin (CR)-binding proteins in a yeast two-hybrid screen utilizing simian cDNA expression-library. The simian CyPA protein had 96% identity with that of human, differing only at eight amino acid residues. We further established CyPA-CR interaction by incubation of glutathione transferase-fused CyPA (GST-CyPA) and CR proteins with CV-1 cyto-lysates, followed by CR and CyPA-specific immuno-blot analysis. The immunosuppressive drug cyclosporin A, a CyPA ligand, did not inhibit CyPA-CR interaction. Our results established a new property of CyPA binding activity to CR. Since CR is a Ca2+-binding protein, CR-CyPA interactions may be important in signaling pathways for induction of Ca2+-dependent cellular processes. PMID- 10456776 TI - Use of fluorescence enhancement technique to study bilirubin-albumin interaction. AB - Bilirubin albumin solution gave an emission spectrum in the wavelength range 500 600 nm with emission maxima at 528 nm when excited at 487 nm. The magnitude of fluorescence intensity increased on increasing bilirubin/albumin molar ratio. At three different albumin concentrations, namely, 1.0, 2.5 and 10.0 microM, there was an initial linear increase in fluorescence up to a molar ratio 1.0 in all cases beyond which it sloped off or decreased. This fluorescence enhancement was used to calculate the binding parameters of bilirubin-albumin interaction and the value of binding constant was found to be 1.72 x 10(7) l/mol similar to the published values obtained with other methods. Different serum albumins, namely, human (HSA), goat (GSA), pig (PSA) and dog serum albumins (DSA) bound bilirubin with almost the same affinity when studied by the technique of fluorescence enhancement. Bilirubin albumin interaction was also studied at different pH and ionic strengths. There was a decrease in bilirubin-albumin complex formation on either decreasing the pH from 9.0 to 7.0 or increasing the ionic strength from 0.15 to 1.0. These results suggest that the technique of fluorescence enhancement can be used successfully to study the bilirubin-albumin interaction. PMID- 10456777 TI - Recent advances in paediatric cardiopulmonary bypass. PMID- 10456778 TI - Retrograde cerebral perfusion: clinical and experimental aspects. PMID- 10456779 TI - Retrograde cerebral perfusion: experimental approach to brain oedema. PMID- 10456780 TI - Biomaterials in cardiopulmonary bypass. PMID- 10456781 TI - Mechanisms of the systemic inflammatory response. AB - The purpose of inflammation is to combat various agents that may injure the tissues. Conditions such as CPB can often cause systemic inflammation and dysfunction of major organs. Pulmonary, renal, myocardial and intestinal function may suffer various degrees of impairment during and after cardiac surgery. Although changes in major organs usually remain clinically insignificant, severe organ failure is not uncommon. The process of systemic inflammation proceeds through activation of serum proteins, activation of leucocytes and endothelial cells, secretion of cytokines, leucocyte-endothelial cell interaction, leucocyte extravasation and tissue damage. Several anti-inflammatory strategies have already been used, some of which have given promising results pertaining to further reduction in the rate of the inflammation-related complications in cardiac surgical patients. PMID- 10456782 TI - Cardiopulmonary bypass for less invasive procedures. PMID- 10456783 TI - Coronary artery bypass graft with biventricular microaxial pumps. PMID- 10456784 TI - Extracorporeal membrane oxygenation. PMID- 10456785 TI - Arteriovenous carbon dioxide removal: development and impact on ventilator management and survival during severe respiratory failure. PMID- 10456786 TI - Progress with the development of the intravenous membrane oxygenator. PMID- 10456787 TI - Inhibition of HIV/SIV replication by dominant negative Gag mutants. AB - There are several major strategies against HIV/AIDS. Of these, the gene therapy is a novel, challenging, and promising one. The target genes, which have been extensively studied for the potential gene therapy of HIV/AIDS, include those of cellular and viral origins. Especially, trans-dominant negative Tat, Rev, Env, Pol, and Gag mutants of HIV have currently attracted considerable attention. In this brief review, we summarize the nature of the HIV/SIV mutants of this category and discuss their future use for gene therapy with special reference to the dominant negative Gag mutants of HIV-1. PMID- 10456788 TI - Sequence analysis of the polymerase domain of HIV-1 reverse transcriptase in naive and zidovudine-treated individuals reveals a higher polymorphism in alpha helices as compared with beta-strands. AB - We report a statistical analysis of genetic heterogeneity of the reverse transcriptase (RT)-coding region of human immunodeficiency virus type 1. Both newly determined sequences and sequences contained in the data banks have been examined. For the calculations, the viral samples and the regions within the RT molecule were divided in two groups. The viral samples were split into those from patients not subjected to antiretroviral therapy and those from patients treated with zidovudine (AZT, 3'-azido-3'-deoxythymidine) alone or in combination with other RT inhibitors. The RT-coding region was divided into segments encoding beta strands and segments encoding alpha-helices. A significantly lower heterogeneity was observed in beta-strands relative to the alpha-helix coding segments. Application of the D test of Tajima has provided evidence of operation of negative (or purifying) selection in sequences from viruses of patients not subjected to antiretroviral treatment as well as in treated patients. In the group of untreated individuals, regions encoding beta-strands are subjected to stronger negative selection than those encoding alpha-helices. It is likely that the observed differences reflect stronger functional constraints in beta-strands than in alpha-helices of RT. PMID- 10456789 TI - Detection of new DNA polymerase genes of known and potentially novel herpesviruses by PCR with degenerate and deoxyinosine-substituted primers. AB - A consensus primer PCR approach was used to (i) investigate the presence of herpesviruses in wild and zoo equids (zebra, wild ass, tapir) and to (ii) study the genetic relationship of the herpesvirus of pigeons (columbid herpesvirus 1) to other herpesvirus species. The PCR assay, based on degenerate primers targeting highly conserved regions of the DNA polymerase gene of herpesviruses, was modified by using a mixture of degenerate and deoxyinosine-substituted primers. The applicability of the modification was validated by amplification of published DNA polymerase genes of 16 herpesvirus species and of the previously uncharacterized DNA polymerase genes of equine herpesvirus 3 (EHV-3) and equine herpesvirus 5 (EHV-5). The modified assay was then used for partial amplification of the polymerase of columbid herpesvirus 1 which is presently classified as a beta-herpesvirus based on biological criteria. Sequence analysis of amplicons obtained from four different viral strains revealed a close relationship of columbid herpesvirus 1 to members of the subfamily Alphaherpesvirinae, especially to Marek's disease herpesvirus. This was confirmed by characterization of additional 1.6kb of the columbid herpesvirus 1 polymerase. Consensus PCR analysis of blood samples from zebras, a wild ass and a tapir revealed amplicons showing high percentages ( > 50%) of sequence identity to DNA polymerases of gamma herpesviruses. In particular, the zebra and the wild ass sequence were closely related to each other and to the polymerases of the equine gamma-herpesviruses EHV-2 and EHV-5 with sequence identities of > 80%. This is a first indication that novel gamma-herpesviruses are present in wild and zoo equids. PMID- 10456790 TI - Molecular evidence for the existence of two distinct subgroups in cucumber mosaic cucumovirus. AB - Infectious full-length cDNA clones from the genomic RNAs of a subgroup II cucumber mosaic cucumovirus strain (Trk7) were obtained. Sequence analysis of the whole genome revealed strong homology (99%) to the genome of Q-CMV, the only subgroup II strain whose entire genomic nucleotide sequence had been available in the database, and an overall 75% homology to those of subgroup I strains. We provide sequence comparisons of different parts of 1a, 2a and 2b proteins of Cucumovirus species, and propose phylogenetic trees based on these protein sequences. PMID- 10456791 TI - Comparisons of nucleotide and deduced amino acid sequences of NSP4 genes of virulent and attenuated pairs of group A and C rotaviruses. AB - The NSP4 protein of rotavirus is a nonstructural glycoprotein and has a crucial function in virus morphogenesis during infection of host cells. It was recently reported that NSP4 may also function as a viral enterotoxin in the induction of rotavirus diarrhea by causing Ca++ influx in the cytoplasm of the infected cells. We sequenced and analyzed two (Wa and M strains) pairs of NSP4 genes of virulent (v) and attenuated (a) (after 30 to 40 passages in cell culture) human group A rotaviruses and a pair of NSP4 genes of virulent and attenuated porcine group C rotavirus (Cowden strain). These strains were previously identified as virulent (induce diarrhea) or attenuated (no diarrhea) in a gnotobiotic pig model of rotavirus infection [Bohl et al. (4), Saif et al. (13), Ward et al. (17)]. The NSP4 genes of the Wa, M and Cowden strains were amplified with RT-PCR using a proof reading polymerase (Tli) and the RT-PCR product was sequenced directly. Analysis of the NSP4 deduced amino acid sequences showed that only 3 (Wa) and 2 (M and Cowden) amino acids differed between the virulent and attenuated strains. For the Wa strain, the changes from the virulent to attenuated strain were in amino acids 13 (V to A), 16 (L to S) and 34 (P to L); in the M strain, the difference was in amino acids 53 (T to I) and 104 (K to E), and in the Cowden strains, amino acids 50 (L to F) and 97 (D to N) differed between virulent and attenuated strains. To our knowledge, this is the first sequence comparison between NSP4 of a virulent and attenuated pair of group C rotaviruses. The potential impact of these few amino acid changes on the pathogenesis of the NSP4 protein for piglets is unclear, relative to previous findings in mice (1), but requires further study using purified recombinant NSP4 proteins or peptides. PMID- 10456792 TI - Polymerase chain reaction amplification and gene sequence analysis of a calicivirus from a feral rabbit. AB - A reverse transcription-polymerase chain reaction (RT-PCR) amplification assay was used to detect calicivirus gene sequences in a liver tissue derived from a feral rabbit which died of a recent outbreak of rabbit haemorrhagic disease (RHD) in New Zealand. Five pairs of primers were designed to amplify five complementary DNA genomic sequence stretching from nucleotide positions 1594 to 7071, yielding amplified fragments of 361, 340, 805,670 and 386 bp for the primer pairs RC-1/RC 2, RC-3/RC-4, RC-5/RC-6, RC-7/RC-8 and RC-9/RC-10 respectively. The identity of the amplified fragments was confirmed by chemiluminescence Southern blot hybridization and direct cycle sequencing. The nucleotide sequences of the five amplified fragments were determined and comparisons of the nucleotide and deduced amino acid sequences revealed a close genetic relationship of the New Zealand isolate 97-10372 with overseas strains of RHD virus. PMID- 10456793 TI - Identification of a gene cluster within the genome of Chilo iridescent virus encoding enzymes involved in viral DNA replication and processing. AB - The nucleotide sequence of the genome of Chilo iridescent virus (CIV) between the genome coordinates 0.974 and 0.101 comprising 27,079 bp was determined. Computer assisted analysis of the DNA sequence of this particular region of the CIV genome revealed the presence of 42 potential open reading frames (ORFs) with coding capacities for polypeptides ranging from 50 to 1,273 amino acid residues. The analysis of the amino acid sequences deduced from the individual ORFs resulted in the identification of 10 potential viral genes that show significant homology to functionally characterized proteins of other species. A cluster of five viral genes that encode enzymes involved in the viral DNA replication was identified including the DNA topoisomerase II (A039L,1,132 amino acids (aa)), the DNA polymerase (ORF A031L,1,273 aa), a helicase (ORF A027L, 530 aa), a nucleoside triphosphatase I (ORF A025L, 1,171 aa), and an exonuclease II (ORF A019L, 624aa), all ORFs possessing the same genomic orientation. The DNA polymerase of CIV showed the highest homology (24.8% identity) to the DNA polymerase of lymphocystis disease virus lymphocystis disease virus 1 (LCDV-1), a member of the family Iridoviridae, indicating the close relatedness of the two viruses. In addition, four putative gene products were found to be significantly homologous to previously identified hypothetical proteins of CIV. PMID- 10456794 TI - Nucleotide sequence of a 5892 base pairs fragment of the LsMNPV genome and phylogenetic analysis of LsMNPV. AB - A 5892 bp fragment of Leucania separata multiple nuclear polyhederosis virus (LsMNPV) containing gp37, 39 k genes and another four ORFs was sequenced in this article. According to regulatory elements on 5' uncoding sequences, the ORF4 and ORF1 are probably two novel baculovirus genes, and the ORF4 probably also is a new early-late gene. The possible functions about GP37 and 39 K proteins were explored. The homology of GP37 protein among LsMNPV and 8 other insect virus was compared. The evolutional position of LsMNPV is also estimated based on the homology of gp37 genes. The structure and its homology of several genes in LsMNPV prove that LsMNPV is far related with other baculovirus and has an exceptional genome organization. PMID- 10456795 TI - Analysis of the sequence of dioscorea Alata bacilliform virus: comparison to others members of the badnavirus group. AB - The complete nucleotide sequence of the genome of Dioscorea alata bacilliform virus (DaBV) has been determined from cloned fragments. Features of the genome confirm DaBV to be a pararetrovirus of the genus Badnavirus which is more similar to other mealy-bug transmitted badnaviruses, in particular to cacao swollen shoot virus, than to rice tungro bacilliform virus. Sequence variability between cloned fragments suggests that the genetic variability of the virus may be quite high (up to 11% nucleotide sequence variation for some small regions of the genome) although the overall variability detected was 4.2% at the nucleotide level. PMID- 10456796 TI - Dissociation between autonomic responding and verbal report in right and left hemisphere brain damage during anticipatory anxiety. AB - OBJECTIVE: To study the relationship between peripheral autonomic arousal and emotional experience. BACKGROUND: Several theories of emotional experience suggest that this experience is, at least in part, related to feedback from the viscera and the autonomic nervous system. METHOD: To partially test this hypothesis and to learn if there are hemispheric asymmetries of emotional experience, we studied skin conductance responses (SCR) and verbal report in patients with right hemisphere damage (RHD), patients with left hemisphere damage (LHD), and normal control subjects during the anticipation of electric shocks. RESULTS: During the shock condition, RHD and LHD subjects had smaller SCRs than did normal control subjects. Verbal report measures, however, revealed that subjects reported feeling less pleasant, more aroused, and less in control during the shock condition compared to the no-shock condition. Unlike the SCR results, the verbal report of emotional experience did not differ between the patients with RHD, patients with LHD, and normal control subjects. CONCLUSIONS: Emotional experience is not dependent upon activation of and feedback from the autonomic nervous system. PMID- 10456797 TI - Anatomical correlates of alien hand syndromes. AB - OBJECTIVE: The objective of this study was to correlate various alien hand syndromes (AHS) with sites of lesion in 16 patients with anterior cerebral artery (ACA) territory infarction. BACKGROUND: All previous reports of AHS were case studies. Their clinico-anatomic correlations were still controversial. While the callosal lesion appears necessary for various types of AHS, which portion of the corpus callosum is associated with which syndrome is still not yet completely resolved. METHOD: Sixteen patients with ACA territory infarction were selected from a stroke registry containing 7355 individuals. They were divided into three lesion groups according to location identified by computed tomography or magnetic resonance imaging: mesial frontal, callosal and combined mesial frontal and callosal. Neurobehavioral examination focused on the presence or absence of grasp reflex, various alien hand signs, intermanual conflict, and callosal disconnection syndrome. By gross visual analysis of the obtained data, a trend for clinicoanatomic correlation emerged. RESULTS: Patients with restricted mesial frontal lesions (three subjects), restricted anterior callosal lesions (four subjects) or "restricted" mesial frontal and anterior callosal lesions (two subjects) did not have symptoms of AHS. The remaining seven patients with extensive callosal injury involving the midbody and isthmus all had symptoms of AHS. Four of the patients with relatively isolated callosal involvement showed intermanual conflict (IMC) associated with a callosal AHS, while the remaining three patients with additional lesions involving the mesial frontal cortex showed impulsive reaching and grasping behaviors of the contralesional hand (a component of frontal AHS) and IMC (a component of callosal AHS). CONCLUSIONS: A comparison of the lesions and symptoms of seven patients in two different groups shows that mesial frontal and anterior callosal (genu and rostral body) lesions are associated with a contralateral frontal AHS. Isolated involvement of the callosal midbody and isthmus is associated with a callosal AHS. When the mesial frontal cortex and corpus callosum (genu and whole body) are extensively involved, a mixed frontal and callosal AHS may occur in the same patient. PMID- 10456798 TI - Frontal lobe lesions and executive dysfunction in children. AB - OBJECTIVE: To investigate the relationship of frontal lobe lesions and neuropsychologic performance in school-aged children to determine whether damage to frontal regions results in specific cognitive sequelae. BACKGROUND: The role of the frontal lobes in executive function remains incompletely understood, particularly in children. METHOD: This retrospective study included children aged 8 to 17 with brain lesions of various etiology (n = 63) or diverse psychiatric disorders (n = 48). All were evaluated for details of neurologic and medical history and for scores on the Wechsler Intelligence Scale for Children (WISC-III) and the Wisconsin Card Sorting Test, and all but the psychiatric patients had neuroimaging scans. Five groups were analyzed--dorsolateral frontal, medial orbital frontal, focal nonfrontal, diffuse, and psychiatric--and neuropsychologic test results were compared using a Kruskal-Wallis nonparametric analysis of variance. RESULTS: Children with damage to dorsolateral frontal regions were more impaired on the Wisconsin Card Sorting Test than those in all other groups. Comparable performance on the Wechsler scale was found in all groups, suggesting that intellectual functioning did not account for this difference. CONCLUSIONS: These data provide evidence for a prominent role of the dorsolateral frontal regions in the mediation of executive function in children. They also support the use of the Wisconsin Card Sorting Test in children as a measure of dorsolateral frontal integrity. PMID- 10456799 TI - Ideomotor apraxia in Alzheimer disease and left hemisphere stroke: limb transitive and intransitive movements. AB - OBJECTIVE: Ideomotor apraxia was studied in patients with Alzheimer disease (AD) and unilateral left hemispheric damaged (LHD) stroke to determine whether these groups differed. BACKGROUND: Given that the neuropathology of AD is bilateral and more diffuse than the localized involvement in patients after an LHD stroke, and given that the cognitive deficits in AD are more widespread than in LHD stroke, the authors predicted that patients with these disorders would differ in response to an auditory command task administered to evaluate ideomotor apraxia, and that the two patient groups would be significantly more impaired than healthy matched control subjects. METHODS: Twenty-one persons were studied, including equal numbers of patients with AD, patients with unilateral LHD stroke, and control subjects. An auditory command test of limb apraxia was administered and videotaped to score performance and to code spatial-temporal or content errors. RESULTS: The patients with AD and LHD stroke were significantly more impaired than healthy control subjects. Whereas the patients with AD and LHD stroke were equally apraxic and did not differ in their performance of transitive limb movements, the patients with AD were significantly more impaired than the patients with stroke when performing intransitive limb movements. A positive correlation was found between severity of dementia and severity of apraxia in the patients with AD. The patients with LHD stroke were as likely to make spatial temporal as content errors when performing intransitive limb movements, whereas the patients with AD made content errors only. Error types produced with transitive limb movements did not differ between groups; spatial-temporal errors were the most common errors made both by patients with AD and patients with LHD stroke. CONCLUSIONS: As predicted, patients with AD and with LHD stroke were impaired when producing limb movements after auditory command, and both patient groups were significantly more impaired than the healthy adults. Patients with AD were significantly more impaired than patients with stroke when performing intransitive limb movements, and error types differed by group. Patients with AD and patients with stroke were equally impaired when performing transitive movements, and error types did not differ by group. Patients with ideomotor apraxia are often degraded in their production of transitive and intransitive movements, and the observation that performance may differ depending on the type of limb movement suggests that movement representations for transitive and intransitive movements may be at least partially independent. PMID- 10456800 TI - Repeated measures of cognitive processing efficiency in adolescent athletes: implications for monitoring recovery from concussion. AB - OBJECTIVE: The objective of this study was to determine whether an adolescent athlete, in the absence of concussion, would be expected to show an improvement in cognitive function during the course of a high school football season. BACKGROUND: At least 60,000 American high school football players suffer cerebral concussion every year, and symptoms may persist for 4 or more years in as many as 24%. METHOD: 34 members of a cohort of healthy athletes, aged 13-18, were administered a computerized neuropsychologic test battery from the Automated Neuropsychological Assessment Metrics (ANAM) before and after the 1997 high school football season, with a mean interval of 16.1 (range 12.3-20.4) weeks between tests. Preseason and postseason scores on eight tests were compared, with significance determined by paired t-test. For those tests in which an improvement was noted, one-way analysis of variance and Wilcoxon tests were used with both preseason and postseason data to determine if there was a measurable difference in cognitive processing efficiency between older and younger subjects. RESULTS: Improvements in processing efficiency (p < 0.001) were noted on tests designed to measure visual scanning and sustained attention (CDS), immediate recall (CDI), and short-term memory (CDD). Older subjects generally performed better on each of these tests, though the difference was significant in only one case (postseason CDI, 17-18 year olds vs. 13-14 year olds, Wilcoxon, p = 0.043). CONCLUSIONS: Our findings suggest that ANAM is sensitive to differences and improvements in cognitive function during a 4 month interval in adolescence. They also suggest that using "return to baseline" cognitive function as the criterion for evidence of recovery from concussion may be insufficient, especially when the baseline measurement was obtained 4 or more months prior to the date of "full recovery." PMID- 10456801 TI - High velocity transient visual processing deficits diminish ability of patients with schizophrenia to recognize objects. AB - OBJECTIVE: Early information processing deficits are consistently reported for patients with schizophrenia. A growing number of studies have applied a transient (magnocellular) or sustained (parvocellular) explanation to account for deficient processing of briefly presented visual stimuli, moving stimuli, and stimuli requiring eye movements in patients with schizophrenia. This reasoning is based on research that makes the distinction between a magnocellular channel, which primarily responds to low spatial frequency and moving or rapidly presented visual information, and a parvocellular channel, which is primarily responsive to high spatial frequency and detailed information. BACKGROUND: Although the preponderance of findings offer support for transient ("where is it") as opposed to sustained ("what is it") deficit in patients with schizophrenia, there remains a need for more specific depiction of the deficit. METHOD: The present study evaluated normal control subjects and patients with schizophrenia recruited from in-patient and out-patient settings. A Motion Defined Letter task was used, owing to its sensitivity to transient (magnocellular) activation. RESULTS: Twenty-three patients with schizophrenia and sixteen normal control subjects were tested on eight dot velocity levels, ranging from 88 arc min/sec to 0.69 arc min/sec. A repeated measures analysis of variance indicated that the performance of patients with schizophrenia was significantly poorer than that of their normal counterparts on the three fastest dot velocity conditions (88 arc min/sec, p < 0.0001, 44 arc min/sec, p < 0.00001, and 22 arc min/sec, p < 0.00003), but performance did not differ on the five slower dot velocity conditions. A regression analysis revealed that the dosage of medication was positively associated with performance on three middle range dot velocity conditions (11 arc min/sec F (1,22) = 6.99; p < 0.025; 5.5 arc min/sec, F (2,20) = 0.379; p = 0.05, and 2.25 arc min/sec F (2,20) = 7.37; p < 0.005). CONCLUSIONS: The findings afford support for an early information processing deficit in schizophrenics. These data also support the neurophysiologic model that explains the poor performance of patients with schizophrenia as it relates to a transient channel deficiency. PMID- 10456802 TI - Cognitive functioning in patients with chronic obstructive pulmonary disease and mild hypoxemia compared with patients with mild Alzheimer disease and normal controls. AB - OBJECTIVE: To examine neuropsychologic functions in patients with chronic obstructive pulmonary disease (COPD) and mild hypoxemia compared with patients with mild Alzheimer disease and normal controls. BACKGROUND: Cognitive deficits have been documented in patients with COPD, but few studies have compared the neuropsychologic status of these patients with that of other neurologic groups. METHOD: Cognitive test results from 32 patients with COPD and mild hypoxemia (mean age, 70.3 years; mean education, 13.2 years; mean partial arterial oxygen pressure, 68.8 mm Hg) who had no neurologic symptoms were compared with 31 subjects with mild Alzheimer disease (AD) and 31 normal controls similar in age, education, and sex. Seventy-three percent of the patients with COPD were receiving supplementary oxygen. RESULTS: Significant group differences across 11 cognitive scores were found using analysis of variance, and post hoc analyses indicated that patients with mild AD performed significantly worse than normal controls and patients with COPD on most tests. The group with COPD and the group with AD demonstrated lower letter fluency compared with controls. Although the patients with COPD performed significantly worse than controls on verbal fluency tasks, they were not in the clinically impaired range, and, overall, the group with COPD was similar to the controls on most cognitive tests. CONCLUSIONS: These findings suggest that many patients with COPD and mild hypoxemia who don't have neuropsychiatric histories may perform normally on cognitive measures. Oxygen therapy may partially account for preservation of cognitive function in these patients. Results also suggest that patients with COPD and normal controls can be readily distinguished from patients with mild AD based on levels and patterns of neuropsychologic test results. Any significant cognitive deficits in patients with mildly hypoxemic COPD may warrant continued neurologic evaluation. PMID- 10456803 TI - Dopamine systems in human immunodeficiency virus-associated dementia. AB - OBJECTIVE: To provide an update of the neurobiologic basis of human immunodeficiency virus (HIV)-associated dementia (HAD), with emphasis on the relationship between dopamine (DA) system dysfunction and behavioral manifestations. BACKGROUND: HIV has a propensity to invade subcortical central nervous system areas, particularly the basal ganglia. Indeed, the core symptoms of HAD are similar to those seen in patients with frontal-striatal dysfunction, the "subcortical dementias" (e.g., Parkinson disease, Huntington disease, progressive supranuclear palsy). FINDINGS: Damage to DA neurons appears to occur in early stages of the disease. Patients with HIV have decreased levels of cerebrospinal fluid DA, and patients with HAD have a reduction of the DA metabolite homovanillic acid but a relative preservation of other neurotransmitters, suggesting a loss of DA neurons. Neuropathologic examinations have shown neuronal loss of the globus pallidus, which is less severe in the neocortex. Furthermore, extrapyramidal signs and marked hypersensitivity to DA antagonists (e.g., neuroleptics) have a propensity to develop in patients with acquired immunodeficiency syndrome. CONCLUSIONS: Neurobiologic investigations suggest that DA system dysfunction plays a critical role in the clinical manifestation of HIV infection, especially HAD. The causes of the vulnerability of this system to the infection are unknown. Understanding this mechanism is important to develop neuroprotective agents in the treatment of HAD and to design new therapies for HAD-related psychiatric symptoms. PMID- 10456804 TI - Autistic regression with rolandic spikes. AB - OBJECTIVE: This study was designed to further define the electroencephalographic abnormalities seen in the Landau-Kleffner syndrome variants and the associated clinical features. BACKGROUND: Landau-Kleffner syndrome is rare, but its putative variants are more common. METHOD: We report two patients with centro-temporal spikes, autistic epileptiform regression, and variably prominent oro-motor symptoms. RESULTS: The epileptic aphasia pattern found among patients with prominent Rolandic spikes may more frequently involve expressive language than is seen in the typical Landau-Kleffner syndrome, where verbal auditory agnosia is the rule. CONCLUSIONS: This clinical difference likely reflects the location of the epileptiform activity (centrotemporal as opposed to anterior or mid-temporal) on buccal-lingual function, vocalization, and language production. PMID- 10456805 TI - The hills are alive with the sound of music. PMID- 10456806 TI - Are we able to interpret the different canary songs? PMID- 10456807 TI - Perioperative gastric tonometric PCO2 and intramucosal pH in patients undergoing liver transplantation. AB - BACKGROUND: Extensive research has focused on the role of insufficient gastro intestinal perfusion and inflammatory activation in the development of organ dysfunction during critical illness. In patients undergoing liver transplantation, portal and caval vein clamping leads to gastro-intestinal and lower extremity venous congestion during the anhepatic phase, and studies suggest that gastro-intestinal perfusion may be compromised. This study was performed to investigate gastro-intestinal perfusion in patients undergoing liver transplantation. METHODS: In 16 patients undergoing liver transplantation, perioperative gastric tonometry with determination of tonometric PCO2, tonometric arterial PCO2 gradient and intramucosal pH were performed. Blood gases were obtained simultaneously from the arterial and portal vein blood. RESULTS: Tonometric PCO2 was 4.6 (4.2/5.3) kPa preoperatively and increased to 5.6 (4.5/6.0) kPa during the anhepatic phase (P<0.01), while the tonometric-arterial PCO2 gradient increased from -0.3 (-0.5/0.0) kPa preoperatively to 0.7 (0.3/1.2) kPa during the anhepatic phase (P<0.01). Intramucosal pH decreased to 7.27 (7.21/7.32) u during the anhepatic phase (P<0.01, compared to preoperatively). The portal vein PCO2 was not significantly different from arterial PCO2 or tonometric PCO2 at any measurement point. CONCLUSION: This study demonstrates that clinical liver transplantation is associated with gastro-intestinal perfusion in the range of aerobic metabolism. The results do not support the presence of gastro-intestinal perfusion in the range of anaerobic metabolism. PMID- 10456808 TI - Intraperitoneal and sigmoid colon tonometry in porcine hypoperfusion and endotoxin shock models. AB - BACKGROUND: The objective of the present study was to assess the accuracy of an air tonometry device in vivo within a wide range of regional carbon dioxide tension (PrCO2) values by using saline tonometry as the standard and to investigate the possibilities to monitor perfusion of the intestine by tonometry in the intraperitoneal cavity. METHODS: Piglets were anesthetized and mechanically ventilated. A pair of tonometry catheters was placed in the sigmoid colon, while another pair was placed intraperitoneally in the right lower quadrant of the abdomen. Air tonometric regional PCO2 (aPrCO2) was measured every 15 min intraperitoneally and every 20 min in the sigmoid colon. Saline tonometric measurements were made every 30 min and steady-state values (ssPrCO2) were derived. Hypoperfusion shock was induced by graded constriction of the aorta. Endotoxin shock was induced by administration of lipopolysaccharide (LPS). RESULTS: The obtained average PrCO2 (=matched (aPrCO2+ ssPrCO2)/2) values ranged from 5.1 kPa to 14.7 kPa. Regional air PCO2 (aPrCO2) and steady-state saline PCO2 (ssPrCO2) exhibited a strong positive linear relationship (r=0.959). The 95% confidence interval of the mean of dPrCO2 (=aPrCO2-ssPrCO2) was 0.31-0.46 kPa. Intraperitoneal tonometric PrCO2 was lower than intraluminal PrCO2 in the sigmoid colon, and was also more sensitive to circulatory changes than sigmoid colon PrCO2. CONCLUSION: The regional air PCO2 (aPrCO2) showed good agreement with the steady-state saline regional PCO2 (ssPrCO2). Intraperitoneal measurements may be an alternative method of monitoring intestinal perfusion after abdominal surgery. PMID- 10456809 TI - Postoperative respiratory arrhythmias: incidence and measurement. AB - BACKGROUND: Postoperative respiratory disturbances may be the result either of depression of respiratory drive, or alteration of respiratory pattern. METHODS: Using an inductance plethysmograph, we continuously recorded the breathing in 52 patients for 4 h after major thoracic, abdominal, and body surface surgery. The ventilatory response to hypercapnia was measured preoperatively, and at the start and end of the observation period. RESULTS: From a variety of statistical measures of respiratory depression, it was found that the occurrence of apnoeas (breath times >10 s), did not correlate with measures of respiratory drive, or with dose or route of administration of opioid, site of surgery, pain, or drowsiness. Instead, the incidence of apnoeas correlated most closely with measures of respiratory pattern (standard deviation of breath times (r=0.72), the mean breath time (r=0.63), approximate entropy of the breath times (r= -0.32)). Twenty-nine percent of patients had some breath times (Ttot) longer than 20 s. CONCLUSION: We would conclude that changes in respiratory pattern are not closely correlated with changes in traditional measures of respiratory drive. PMID- 10456810 TI - Effects of dopamine in lung-transplanted pigs at 32 degrees C. AB - BACKGROUND: We have used hypothermia successfully in patients with acute respiratory failure after lung transplantation. However, we have observed that dopamine may cause a substantial decrease in mean arterial pressure (MAP) in hypothermic subjects. Furthermore, a dopamine-induced increase in pulmonary vascular resistance (PVR) has been reported in the literature, and this could aggravate the increase in PVR which is often seen both in the early postoperative phase after lung transplantation, and during hypothermia. We thus hypothesized that dopamine would decrease MAP and increase PVR in hypothermic lung transplanted subjects. METHODS: Left single lung transplantation combined with right pneumonectomy was performed in 6 pigs anesthetized with ketamine and midazolam and muscle relaxed with pancuronium. After an observation period of 24 h, the effect of dopamine infused at 5 and 12 microg x kg(-1) x min(-1) was studied in normothermia (38 degrees C) and after cooling by cold-water immersion to 32 degrees C. RESULTS: Systemic vascular resistance index (SVRI) increased and cardiac index (CI) decreased in hypothermia. Dopamine decreased SVRI and increased CI, both in normothermia and at 32 degrees C. MAP decreased during infusion of dopamine in hypothermia. Dopamine had no effect on the pulmonary vascular resistance index (PVRI). CONCLUSION: The results suggest that dopamine may be used to increase cardiac output after lung transplantation in moderate hypothermia as well as in normothermia, but one should be aware that dopamine may cause a substantial decrease in blood pressure, depending on the prevailing hemodynamic conditions at the start of its administration. PMID- 10456811 TI - Dopamine reduces gastric tone in a dose-related manner. AB - BACKGROUND: Dopamine may have effects on gastrointestinal motility. The aim of this study was, therefore, to determine whether dopamine reduces gastric tone and whether the effects of dopamine can be blocked by a dopamine antagonist. METHODS: Eight healthy male volunteers were studied on two occasions in a randomized order. A continuous infusion of incremental doses of dopamine (2.5, 5.0, 7.5 microg kg(-1) min(-1)) was given on both occasions. Ten milligrams of the dopamine-antagonist metoclopramid was given before the dopamine infusion on one occasion and during the dopamine infusion (7.5 microg kg(-1) min(-1)) on the other occasion. The gastric tone was measured by an electronic barostat, an instrument with an electronic control system that maintains a constant preset pressure within an air-filled, flaccid intragastric bag by means of momentary changes in the intragastric volume of air. Volume and pressure in the gastric bag were continuously recorded by the electronic barostat and sampled in a computer. RESULTS: Dopamine induced a dose-related increase in the intragastric bag volume. Metoclopramid given as a 10 mg i.v. bolus dose during the infusion of dopamine significantly decreased the intragastric bag volume, but 10 mg of metoclopramid i.v. before the dopamine infusion did not influence the bag volume per se. CONCLUSION: Dopamine decreases gastric tone in a dose-related manner and 10 mg of the dopamine-antagonist metoclopramid is not enough to fully reverse these effects. PMID- 10456813 TI - Perioperative transcutaneous pacemaker in patients with chronic bifascicular block or left bundle branch block and additional first-degree atrioventricular block. AB - BACKGROUND: Complete heart block is dreaded perioperatively in patients with chronic bifascicular or left bundle branch block (LBBB) and additional first degree A-V block. Our aim was to investigate the necessity as well as the efficacy and safety of transcutaneous pacing in the perioperative setting. METHODS: Thirty-nine consecutive patients with asymptomatic chronic bifascicular block or LBBB and prolongation of the P-R interval scheduled to undergo surgery under anesthesia were prospectively enrolled in the study. Preoperatively, a transcutaneous pacemaker (PACE 500 D, Osypka Co.) was applied; its efficacy was checked with intra-arterial blood pressure measurement; the pain level was recorded. Additionally, 24-h Holter monitoring (CM2, CM5) was applied. Occurrences of a block progression or a bradycardia of <40 beats/min with hemodynamic impairment were the defined end points. RESULTS: Thirty-seven of the 39 patients (95%) could be successfully stimulated with a median current strength of 70 mA; whereby 33 of the 39 patients felt moderate to severe pain. There was no perioperative block progression. Three cases of brady-cardia of <40 beats/min with a critical drop in blood pressure occurred; but these patients were successfully treated with drug therapy without pacemaker stimulation. CONCLUSION: The perioperative application and testing of the pacemaker was safe and could be performed in nearly all patients successfully. However, we do not consider a routine prophylactic transcutaneous placement in patients with chronic bifascicular or LBBB and additional first-degree A-V block justified. Nevertheless, appropriate drugs and temporary pacemaker equipment should be easily accessible. PMID- 10456812 TI - Factors affecting gentamicin pharmacokinetics in septic patients. AB - BACKGROUND: This prospective, comparative study was designed to estimate the volume of distribution (Vd) and elimination rate constant (K(e)) of gentamicin and to determine the clinical factors affecting the pharmacokinetics of gentamicin in different stages of sepsis. METHOD: Seventy-seven critically ill patients treated with gentamicin for gram-negative sepsis were included. These septic patients were divided into hyperdynamic septic and hypodynamic septic groups according to cardiac index. Twenty-seven patients who received postoperative prophylactic gentamicin were recruited as controls. RESULTS: Fifty two patients in the hyperdynamic septic group had a significantly larger Vd than those in the hypodynamic septic and control groups. The Vd was correlated significantly with both Acute Physiological Score (APS) (r=0.340, P<0.01) and cardiac index (r=0.394, P<0.01). The K(e) of gentamicin correlated significantly with both blood urea nitrogen (BUN) (r= 0.565, P<0.01) and serum creatinine level (r=0.563, P<0.01). CONCLUSION: The increased Vd in the septic patients was related to the severity of illness and magnitude of cardiac output. The K(e) of gentamicin was correlated with the serum creatinine level. PMID- 10456814 TI - Does desflurane alter left ventricular function when used to control surgical stimulation during aortic surgery? AB - BACKGROUND: Although desflurane is commonly used to control surgically induced hypertension, its effects on left ventricular (LV) function have not been investigated in this clinical situation. The purpose of the present study was to evaluate the LV function response to desflurane, when used to control intraoperative hypertension. METHODS: In 50 patients, scheduled for vascular surgery, anesthesia was induced with sufentanil 0.5 microg/kg, midazolam 0.3 mg/kg and atracurium 0.5 mg/kg. After tracheal intubation, anesthesia was maintained with increments of drugs with controlled ventilation (N2O/O2=60/40%) until the start of surgery. A 5 Mhz transesophageal echocardiography (TEE) probe was inserted after intubation. Pulmonary artery catheter and TEE measurements were obtained after induction (to)(control value), at surgical incision (t1) if it was associated with an increase in systolic arterial pressure (SAP) greater than 140 mmHg (hypertension) and after control of hemodynamic parameters by administration of desflurane (return of systolic arterial pressure to within 20% of the control value) (t2) in a fresh gas flow of 31/ min. RESULTS: Sixteen patients developed hypertension at surgical incision. SAP was controlled by desflurane in all 16 patients. Afterload assessed by systemic vascular resistance index (SVRI), end-systolic wall-stress (ESWS) and left-ventricular stroke work index (LVSWI) increased with incision until the hypertension returned to post induction values with mean end-tidal concentration of 5.1+/-0.7% desflurane. No change in heart rate, cardiac index, mean pulmonary arterial pressure, stroke volume, end-diastolic and end-systolic cross-sectional areas, fractional area change and left ventricular circumferential fiber shortening was noted when desflurane was added to restore blood pressure. CONCLUSION: This study demonstrates that in patients at risk for cardiac morbidity undergoing vascular surgery, desflurane is effective to control intraoperative hypertension without fear of major cardiac depressant effect. PMID- 10456815 TI - Ketamine antagonises alfentanil-induced hypoventilation in healthy male volunteers. AB - BACKGROUND: The effects of ketamine on respiration, alone, or in combination with opioids, have not been completely clarified. Both stimulant and depressant effects have been reported, as well as attenuation of opioid-induced hypoventilation at the expense of increased oxygen consumption. These conflicting results might partly be due to dose-dependent mechanisms. We have, therefore, determined the ventilatory effects of ketamine, in combination with alfentanil, using infusions to different pseudo steady-state concentrations. METHODS: On two separate days, eight healthy male volunteers were given alfentanil as a continuous computer-controlled infusion, aiming at a plasma concentration of 50 ng x mL(-1). After reaching apparent steady-state for alfentanil, racemic ketamine or placebo was administered in a protocol randomised for the two days. On the ketamine days a computer-controlled infusion, aiming for escalating ketamine plasma concentrations of 50, 100 and 200 ng x mL(-1), was added to the alfentanil infusion. On the placebo days saline was added. Using a face-mask with an occlusion valve, respiratory parameters were measured during air-breathing and after 6 repetitive 30-s CO2 challenges. RESULTS: The alfentanil infusion induced hypoventilation by decreasing respiratory rate, while tidal volume and respiratory drive were unaffected. This hypoventilation was antagonised by ketamine in a concentration-dependent manner mainly through an increase in respiratory rate. The CO2 response was not affected by alfentanil or ketamine. CONCLUSION: In the dose range of interest for postoperative, intensive-care and pain-clinic settings, ketamine antagonises the resting hypoventilation induced by alfentanil. PMID- 10456816 TI - Intrathecal co-administration of NMDA antagonist and NK-1 antagonist reduces MAC of isoflurane in rats. AB - BACKGROUND: Intravenous administration of N-methyl-D-aspartate (NMDA) receptor antagonists and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor antagonists reportedly reduce the minimum alveolar anaesthetic concentration (MAC) for inhalation anaesthetics. If pain perception can be prevented by the intrathecal administration of antinociceptive receptor antagonists, these agents may reduce the requirements for inhalation anaesthetics. We studied the effect of intrathecal administration of an AMPA/kainate receptor antagonist, a metabotropic glutamate (mGlu) receptor antagonist and co-administration of NMDA and a neurokinin-1(NK-1) receptor antagonist drugs at low doses on the MAC. METHODS: After Wistar rats (n=36) were fitted with indwelling intrathecal catheters, the MAC of isoflurane was determined following intrathecal administration of a non-NMDA receptor antagonist (CNQX) at 10 microg, a mGlu receptor antagonist (AP3) at 10 microg, or a combination of NMDA receptor antagonist (APV) at 0.01 microg to 1 microg with NK 1 receptor antagonist (CP96345, CP) at 0.1 microg to 10 microg. Subsequently, a reversal dose of intrathecal NMDA with substance P (SP) was administered, and the MAC of isoflurane was redetermined. Conscious rats (n=15) were also examined for the presence of locomotor dysfunction following the intrathecal co-administration of APV and CP. RESULTS: Neither CNQX nor AP3 reduced the MAC of isoflurane. APV at 0.01 microg plus CP at 1 microg, as well as APV at 0.1 microg plus CP at 10 microg, reduced the MAC of isoflurane, with respective reductions of 7.6% and 14%; (P<0.05). Co-administration of NMDA plus SP reversed the decrease in the MAC of isoflurane. Locomotive activity was not changed. CONCLUSIONS: The NMDA receptor and the NK-1 receptor are important determinants of the MAC of isoflurane, exerting this influence by inhibition of pain transmission in the spinal cord, while mGlu and AMPA receptors have no effect on the MAC of isoflurane. PMID- 10456817 TI - Halothane decreases bacterial adherence in vitro. AB - BACKGROUND: Adherence of pathogenic bacteria to host epithelial cells is thought to be the initial step in infection, while the presence of the commensal flora is an important host defence mechanism. Anything altering bacterial adherence to human epithelial cells may contribute to bacterial infections. The impact of anaesthesia on this first step to infection is not known. In this study the effect of halothane on bacterial adherence was investigated. METHODS: Human epithelial cells (HEp-2) and two strains of Escherichia coli were exposed to halothane 2% for 2 h. Then HEp-2 cells were coincubated with bacteria for 3 h. Bacteria attached to the epithelial cells were evaluated by light microscopy. RESULTS: Compared to the control, bacterial adherence was reduced by 37% to 56% with the different strains when HEp-2 cells were exposed to halothane. No significant difference was found when only bacteria were treated with halothane. CONCLUSION: Our results show that halothane reduces bacterial adherence to human epithelial cells in vitro. Reduced number or function of epithelial cell surface receptors may be responsible for the reduced adherence as no changes were observed when only the bacteria were exposed to halothane. PMID- 10456818 TI - Caffeine- or halothane-induced contractures of masseter muscle are similar to those of vastus muscle in normal humans. AB - BACKGROUND: Skinned fibers from normal human masseter muscle have greater caffeine and calcium sensitivity than skinned fibers from vastus muscle. We examined sensitivity to caffeine and halothane in fresh, cut muscle bundles (non skinned) from human masseter muscle. METHODS: Masseter bundles (caffeine, n=25, halothane, n=19) excised from 10 humans under general anesthesia had tension measured in 37 degrees C baths during the addition of caffeine (0.5, 1, 2, 4, 8, 32 mM) or 3% halothane. Results were compared to those of our previous studies (1989, 1997, 25 patients) of vastus bundles (caffeine, n=71, halothane, n=63) using the same protocol, technicians, and equipment. RESULTS: Baseline force in the caffeine test was 2.10+/-1.57 for masseter, and 2.02+/-1.68 and 1.82+/-1.29 respectively for vastus muscle. Force at 32 mM caffeine concentration was 11.2+/ 9.9 g for masseter, 11.0+/-5.4 and 13.5+/-7.5 g for vastus. Concentration response curves were virtually identical. In the halothane group, neither baseline values (masseter 1.47+/-1.30, vastus 1.91+/-1.32 and 2.15+/-1.71) nor contractures in response to 3% halothane were different. Most bundles had no contracture in response to 3% halothane; 3 masseter bundles and 2 vastus bundles (1989) developed contractures of less than 0.05 g. Three vastus bundles (1997) developed contractures >0.2 g. CONCLUSION: Contracture responses of intact cut masseter and vastus bundles (non-skinned) do not differ with respect to caffeine and halothane. Responses of skinned fibers might demonstrate greater sensitivity under certain conditions, but they do not reflect those of intact cut bundles. PMID- 10456819 TI - Single-injection paravertebral block compared to general anaesthesia in breast surgery. AB - BACKGROUND: Breast surgery is frequently associated with post-operative nausea, vomiting, pain and painful restricted movement. Paravertebral block may be an alternative to general anaesthesia for this type of surgery. We studied the single-injection paravertebral block at the level of T4 and report a comparison of single-injection paravertebral block to general anaesthesia for breast surgery. METHODS: After written informed consent was obtained, 86 patients were enrolled in this prospective study. Forty-four women were randomly allocated to receive a single-injection paravertebral block at the level of T4, while 42 women received general anaesthesia. The surgical procedures varied from lumpectomy (wide local excision of a tumour) to modified radical mastectomy with axillary dissection. The block was performed according to the guidelines described by Eason and Wyatt using 0.3 ml x kg(-1) (maximum dose 150 mg) of bupivacaine 0.5%. The skin and the underlying tissues were infiltrated with local anaesthetic solution two fingers (about 3 cm) from the anatomical midline and level with the cephalad end of the vertebral spine. RESULTS: Time for performance of blocks lasted from 4 to 9 min. Recovery from anaesthesia or sedation was shortened, while postoperative pain scores (VAS), the incidence of vomiting and the requirement for analgesics were lower in the paravertebral group. Less painful restricted movement was observed in the paravertebral block group. Paravertebral block was inadequate in 6.8% of patients. Epidural spread with paraparaesis and Horner triad was assumed in one patient. CONCLUSION: Single-injection paravertebral block at the level of T4 represents a suitable alternative to general anaesthesia in women undergoing breast surgery. PMID- 10456821 TI - Gastric intramucosal pH as a monitor of gut perfusion after thrombosis of the superior mesenteric vein. AB - Gastric intramucosal pH (pHi) when measured by a tonometer is a simple and minimally invasive method to determine gut ischemia. In a case of severe mesenteric venous thrombosis, we measured pHi intra- and postoperatively over a period of five days. The goal was to monitor improvement or deterioration of gastrointestinal perfusion in the intensive care unit and to perform a second look laparotomy if the condition worsened. We observed that gastric pHi is a more sensitive parameter for detecting intestinal ischemia than parameters such as arterial pH, base excess, or lactate. This patient's pHi rose continuously, which allowed us to proceed in a conservative way without any further invasive diagnostic interventions. Thus, the application of a gastric tonometer in cases of mesenteric venous thrombosis may help to reduce costs by preventing unnecessary postoperative diagnostic maneuvers such as angiography, computed tomography, or even second-look laparotomy. PMID- 10456820 TI - Intravenous ketoprofen and epidural sufentanil analgesia in children after combined spinal-epidural anaesthesia. AB - BACKGROUND: Epidural opioid analgesia has become more popular for postoperative pain treatment in children. Epidural opioids are associated with adverse effects such as respiratory depression, excessive sedation, protracted vomiting, urinary retention and pruritus. Following minor surgery, ketoprofen has a synergistic effect with opioids, resulting in an improved analgesia without increase in incidence of adverse effects. To see whether this is also true following major surgery, we compared the effect of i.v. ketoprofen and placebo as an adjuvant to epidural sufentanil analgesia. METHODS: A prospective, randomised, double-blind, placebo-controlled, parallel-group study design was used in 58 children, aged 1 15 years, receiving a standardised combined spinal-epidural anaesthesia. Intravenous ketoprofen or saline was provided as a bolus and a continuous infusion in addition to epidural sufentanil infusion, which was adjusted as clinically required. Epidural bupivacaine was used for rescue analgesia. The study drug infusion was discontinued when pain scores were <3 on a 0-10 scale for 6 h with an epidural sufentanil infusion rate of 0.03 microg kg(-1) h(-1). RESULTS: Children in the ketoprofen group received less rescue analgesia (none/29 vs. 8/29 children in the placebo group). In the ketoprofen group, criteria to discontinue epidural sufentanil were achieved more often (14 vs. 6 children) before the end of the 72 h study period. Less children in the ketoprofen group suffered pruritus (13 vs. 4). The incidence of nausea/retching and vomiting was similar (11 vs. 12) in both groups. CONCLUSION: In this study, ketoprofen as a background analgesic to epidural sufentanil provided improved postoperative analgesia and reduced incidence of adverse effects of the epidural opioid. PMID- 10456822 TI - Life-threatening haemorrhage following obturator artery injury during transurethral bladder surgery: a sequel of an unsuccessful obturator nerve block. AB - In spite of prior blockade of the obturator nerve with 1% mepivacaine (8 ml) utilizing a nerve stimulator, violent leg jerking was evoked during transurethral electroresection of a bladder tumour approximately 1 h after the blockade in a 68 year-old man. The patient became severely hypotensive immediately following the jerking, and a large lower abdominal swelling concurrently developed. The urgent laparotomy indicated that the left obturator artery was severely injured by the resectoscope associated with the bladder perforation, causing acute massive haemorrhage. The patient recovered uneventfully after adequate surgery. Investigation of the literature suggested that both our nerve stimulation technique and anatomical approach were appropriate. It was therefore unlikely that our block resulted in failure because of an inappropriate site for deposition of the anaesthetic. However, consensus does not appear to have been obtained as to the concentration and volume of the anaesthetic necessary for prevention of the obturator nerve stimulation during the transurethral procedures. The concentration and volume of mepivacaine we used might have been too low and/or small, respectively, to profoundly block all the motor neuron fibres of the nerve. Alternatively, stimulation of the obturator nerve might occur because of the presence of some anatomical variant, such as the accessory obturator nerve or its abnormal branching. In conclusion, some uncertainty appears to exist in the effectiveness of the local anaesthetic blockade of the obturator nerve. In order to attain profound blockade of the motor neuron fibres of the obturator nerve and thereby prevent the thigh-adductor muscle contraction which can lead to life-threatening situations, we recommend, even with a nerve stimulator, to use a larger volume of a higher concentration of local anaesthetic with a longer duration in the obturator nerve block for the transurethral procedures. PMID- 10456823 TI - Matching appropriately sized reinforced laryngeal mask airways with Boyle-Davis gags for paediatric adenotonsillectomies. PMID- 10456824 TI - Nasogastric tube knotting in open heart surgery. PMID- 10456825 TI - [Management of complicated scar hernias]. PMID- 10456826 TI - [Baker's cyst]. PMID- 10456827 TI - Audit and quality control in angioplasty in Europe: procedural results of the AQUA Study 1997: assessment of 250 randomly selected coronary interventions performed in 25 centres of five European countries. AQUA Study Group, Nucleus Clinical Issues, Working Group Coronary Circulation, of the European Society of Cardiology. AB - AIMS: Percutaneous transluminal coronary angioplasty (PTCA) has become the most widely used major intervention in western medicine. However, there is disparate use of this technique among different European countries and the U.S.A. In an attempt at quality assurance, the working group Coronary Circulation of the European Society of Cardiology has carried out a study on appropriateness, necessity, and performance of PTCA in Europe. The present paper reports on the procedural results of this survey. METHODS: In a multicentre case-control study in Europe, 750 patients (544 men, 206 women) with documented vascular disease of the From the countries participating in the European Registry of Coronary Intervention, the three countries with the highest absolute PTCA volume (Germany, France, and the United Kingdom) and two randomly selected countries (Belgium and Italy) were chosen for investigation. In these countries, five centres were selected at random according to the following criteria: one centre with >1000, three centres with 300-1000, and one centre with <300 procedures per year. In each of these, 10 cases from the first half of 1997 were randomly identified and all pertinent documentation was collected. RESULTS: In 250 cases, 325 stenoses were addressed as target lesions. Single vessel disease was present in 41%. History included stable angina in 49%, unstable angina in 32%, atypical chest pain in 6%, no anginal pain in 12%, and acute/subacute myocardial infarction in 13%. The percentage of patients with either positive stress test and/or unstable angina, acute/subacute infarction, previous infarction (within 6 months) or coronary revascularization amounted to 98%. Single vessel intervention accounted for 90%. In 41% balloon-only angioplasty was performed and in 54% at least one stent was implanted with considerable variation among countries. The use of other new devices amounted to only 3%. In 92%, the operators documented a successful procedure. Major complications (myocardial infarction, emergency bypass surgery, or death) were found in 4.8%. CONCLUSIONS: Based on scrutinized hospital and operator data, the present study revealed a satisfactorily high percentage of justifiable indications, an adequate procedural success rate, and an acceptably low complication rate. Further analysis by an expert panel will address appropriateness, necessity, and procedural performance of the individual cases. PMID- 10456828 TI - 26-cholesterol hydroxylase in rat corpora lutea: A negative regulator of progesterone secretion. AB - From a subtracted cDNA library of rat luteal tissue, where cDNA fragments in functional luteal tissue were subtracted from those in regressing luteal tissue, a cDNA clone corresponding to 26-cholesterol hydroxylase (P450(C26)) was obtained. It is known that P450(C26) catalyzes the conversion of cholesterol to 26-hydroxycholesterol, which blocks cholesterol utilization in the cell, and that 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD) catalyzes the conversion of progesterone to an inactive steroid, 20alpha-dihydroprogesterone (20alpha-OHP). Thus, using pseudopregnant rats as a model, physiological cooperation of P450(C26) and 20alpha-HSD in the reduction of progesterone release toward the end of the luteal phase was evaluated. Levels of P450(C26) and 20alpha-HSD mRNA were examined in corpora lutea from pseudopregnant rats by Northern blot or reverse transcription-polymerase chain reaction or both. P450(C26) mRNA was ubiquitously expressed in corpora lutea, and its expression increased toward the end of pseudopregnancy, while 20alpha-HSD was expressed in all corpora lutea on Day 16 (Day 0 = the day of after cervical stimulation) but not detected before Day 10. An inhibitor of 20alpha-HSD, STZ26 (D-homo-16-oxa-4-androstene-3,16alpha-dione), was administered at various doses to rats from Day 12 to 20, effectively suppressing the elevation of 20alpha-OHP in a dose-dependent manner but not the depletion of progesterone completely. The expression of P450(C26) mRNA was increased as STZ26 dose increased, which negatively correlated with the progesterone levels. These results strongly suggest that P450(C26) cooperated with 20alpha-HSD in the reduction of progesterone release from the rat luteal tissue at the end of the functional luteal phase. PMID- 10456829 TI - Cellular source in ewes of prostaglandin-endoperoxide synthase-2 in uterine arteries following stimulation with lipopolysaccharide. AB - Prostaglandin-endoperoxide synthase (PTGS) (also known as cyclooxygenase) converts arachidonic acid into several prostaglandins, many of which have roles in vasodilation and vasoconstriction under normal and pathological conditions. There are two isoforms of PTGS: PTGS-1 and PTGS-2; PTGS-1 is constitutively expressed in many tissues and is believed to be involved in the homeostatic maintenance of the body. In contrast, PTGS-2 is believed to have a "differentiative" role in the cells and is highly inducible during inflammation and in response to lipopolysaccharide (LPS). Endothelial cells as well as vascular smooth muscle cells can be a source of PTGS within the artery. The objective of this study was to determine the cell population(s) in uterine arteries that respond to LPS with an increase in PTGS-2 protein expression. Uterine arteries collected from ewes during the follicular (Day 0, Day 0 = estrus, n = 4) or luteal (Day 10, n = 4) phase were treated in vitro with LPS as intact artery segments, cut-open artery segments, or cut-open and denuded (endothelial cells absent) artery segments. After 24 h of LPS treatment, intact, cut-open, and denuded uterine artery segments were collected into homogenization buffer for determination of PTGS-2 protein levels by Western blot analysis. The culture medium was collected and used for detection of 6-keto-prostaglandin F(1alpha) (6-keto-PGF(1alpha)), the stable metabolite of prostacyclin, using an enzyme immunoassay. In addition, the location of PTGS-2 after LPS treatment was analyzed by immunohistochemistry in intact artery segments. Denuded arteries (endothelium absent) did not show increases in PTGS-2 protein in the homogenates or 6-keto-PGF(1alpha) in the culture medium after LPS exposure. In contrast, cut uterine arteries responded to LPS stimulation with a significant increase in PTGS 2 protein in homogenates and 6-keto-PGF(1alpha) in culture medium. Immunohistochemical staining for PTGS-2 was associated with both endothelial cells and vascular smooth muscle cells. These results suggest that while both endothelial cells and vascular smooth muscle cells are associated with PTGS-2, after LPS exposure it is the endothelial cells that are essential in uterine artery increases in PTGS-2 and prostacyclin in response to LPS stimulation. PMID- 10456830 TI - Oocytectomy does not influence synthesis of hyaluronic acid by pig cumulus cells: retention of hyaluronic acid after insulin-like growth factor-I treatment in serum-free medium. AB - Mouse oocytes secrete a factor that enables cumulus cells to undergo expansion in response to FSH (1 microg/ml), whereas expansion of the porcine cumulus oophorus has been shown to be independent of the oocyte. The aim of this study was to assess FSH-induced synthesis of hyaluronic acid (HA) by porcine cumulus cells before and after oocytectomy. In addition, we studied the effect of insulin-like growth factor-I (IGF-I) on the ability of cumulus cells to synthesize and retain HA in response to FSH in serum-free medium. Porcine oocyte-cumulus complexes and complexes from which the oocytes had been removed by oocytectomy were cultured for 24 h in the presence of 2.5 microCi of D-[6-(3)H]glucosamine hydrochloride, fetal calf serum (FCS, 5%), and FSH. After 24 h, incorporation of [(3)H]glucosamine into HA was measured either in complexes alone (retained HA) or in medium plus complexes (total HA). Specificity of incorporation of radioactivity into HA was confirmed by the sensitivity to highly specific Streptomyces hyaluronidase. Our results suggest that 1) the synthesis of HA by pig cumulus cells in vitro is stimulated by FSH and that oocytectomy does not change this synthesis; 2) oocytes do not influence retention of HA within the complex; 3) FSH-induced synthesis of HA by cumulus cells is decreased in medium with polyvinylpyrrolidone (PVP)-supplemented (total and retained HA) compared to FCS-supplemented medium; 4) IGF-I enabled cumulus cells to synthesize HA in response to FSH in PVP-supplemented medium in a manner similar to that observed when serum is present in the medium. PMID- 10456831 TI - Nitric oxide regulates human sperm capacitation and protein-tyrosine phosphorylation in vitro. AB - The aim of the present study was to investigate whether the generation of nitric oxide by human spermatozoa is associated with human sperm capacitation and with the tyrosine phosphorylation of sperm proteins. Human spermatozoa were capacitated in the presence or absence of nitric oxide-releasing compounds or nitric oxide synthase inhibitors, and then the percentage of acrosome loss induced by human follicular fluid or by calcium ionophore was determined. The presence of the nitric oxide-releasing compounds primed spermatozoa to respond earlier to human follicular fluid whereas nitric oxide synthase inhibitors decreased the percentage of acrosome reaction. Moreover, nitric oxide modulated tyrosine phosphorylation of sperm proteins. A tight correlation between capacitation and tyrosine phosphorylation regulated by nitric oxide was observed. Results indicate that nitric oxide is involved in human sperm capacitation and emphasize the importance of oxidoreduction reactions in the fine control of sperm physiology. PMID- 10456832 TI - Magnetic cell sorting is a fast and effective method of enriching viable spermatogonia from Djungarian hamster, mouse, and marmoset monkey testes. AB - Germ cell transplantation, which offers promising new approaches for research and clinical applications, has focused interest on spermatogonia. This paper describes a procedure that permits the isolation of large quantities of viable spermatogonia. The immunomagnetic isolation procedure was applied to testicular cell suspensions from photoinhibited and photostimulated Djungarian hamsters, mice, and marmoset monkeys. The cells were incubated with a polyclonal rabbit anti-c-kit IgG, binding of which was characterized by immunohistochemical staining. For magnetic labeling, a secondary anti-rabbit IgG conjugated to ferromagnetic microbeads was used. Separation columns allowed the retention of magnetically labeled cells within the matrix. The magnetic fractions were eluted after removal of the column from the magnetic field. All fractions were analyzed for cellular morphology and by flow cytometry. The final enrichment of c-kit positive cells in the magnetic fraction using fully active testes was in the range of 25-55% with a viability rate of 80-90%. The magnetic fractions of all three species were characterized by high numbers of diploid cells. Cytological analysis revealed a strong enrichment of spermatogonia. No haploid cells were retained in the magnetic fraction. In comparison to conventional procedures, magnetic cell separation is an efficient and fast approach for isolation of spermatogonia. PMID- 10456833 TI - Bovine seminal plasma phospholipid-binding proteins stimulate phospholipid efflux from epididymal sperm. AB - Several studies have shown that sperm capacitation was accompanied by a change in the lipid composition of the sperm membrane. In cattle, the major proteins of (bovine)seminal plasma (BSP proteins: BSP-A1/A2, BSP-A3, and BSP-30-kDa) potentiate sperm capacitation induced by high-density lipoprotein (HDL). Our recent studies indicate that these proteins and HDL stimulate sperm cholesterol efflux during capacitation. In order to gain more insight into the mechanisms of BSP-mediated sperm capacitation, we studied whether or not BSP proteins induce phospholipid efflux from epididymal sperm membrane. By direct determination of choline phospholipids on unlabeled epididymal sperm, the results show that sperm incubated in the presence of BSP-A1/A2 protein lost 34.4% of their choline phospholipids compared with the control (11.5%). Similar results were obtained using labeled epididymal sperm. Labeling was carried out by incubating washed epididymal sperm for 1 h with medium containing [(3)H]palmitic acid. The majority of the label was incorporated into sperm phosphatidylcholine. Studies of sperm phospholipid efflux were done by incubating the labeled sperm with purified BSP proteins, delipidated BSA, or bovine seminal ribonuclease (RNase, control protein). When labeled ([(3)H]phospholipid) epididymal sperm were incubated with BSP proteins (20-120 microg/ml) for 8 h, the sperm lost [(3)H]phospholipid in a dose-dependent manner (maximum efflux of approximately 30%). After the incubation with BSP proteins, the efflux particles were fractionated by size-exclusion chromatography. Analysis of the fractions obtained showed that the [(3)H]phospholipid was associated with BSP proteins. BSA (6 mg/ml) stimulated a specific phospholipid efflux of approximately 22%. In contrast, bovine RNase (120 microg/ml) did not stimulate phospholipid efflux. These results indicate that BSP proteins participate in the sperm cholesterol and phospholipid efflux that occurs during capacitation. PMID- 10456834 TI - Prenatal testosterone masculinizes synaptic input to gonadotropin-releasing hormone neurons in sheep. AB - In sheep, the control of tonic and surge GnRH secretion is sexually differentiated by testosterone in utero. However, GnRH neurons are not sexually dimorphic with respect to number, distribution, or gross morphology. Therefore, this study tested the hypothesis that prenatal steroids influence synaptic input to GnRH neurons. We compared the number of synapses on GnRH neurons from male, female, and androgenized female lambs (n = 5 each). Androgenized females were exposed to testosterone during mid-gestation. Yearling lambs were perfused, and GnRH neurons were visualized using the LR-1 antibody. Five to seven GnRH neurons from the rostral preoptic area in each animal were viewed at the ultrastructural level. Afferent synapses and glial ensheathment on each neuron were counted in a single section through the plane of the nucleus. GnRH neurons from females received approximately twice as many contacts (3.6 +/- 0.7 synapses/100 microm plasma membrane) as those from male lambs (1.6 +/- 0.3; p < 0.05), similar to previous reports in rats. In addition, the number of synapses on GnRH neurons from androgenized female lambs (1.5 +/- 0.5) was similar to that from male lambs, suggesting that prenatal steroids give rise to sex differences in synaptic input to GnRH neurons. PMID- 10456835 TI - Infertility in female rabbits (Oryctolagus cuniculus) alloimmunized with the rabbit zona pellucida protein ZPB either as a purified recombinant protein or expressed by recombinant myxoma virus. AB - Development of immunocontraceptives for wild rabbit populations requires selection of both effective antigens and effective delivery systems. Recombinant rabbit zona pellucida glycoprotein B (ZPB) produced in eukaryotic cells in vitro was an effective antigen and induced sustained infertility in 70% of female rabbits. This required two boosts and serum antibody titers of 12 800 or greater. Antibody titers in females were low after the initial immunization, as might be expected with a self-antigen; however, male rabbits had a strong antibody response, indicating that the protein was immunologically foreign. To develop a delivery system, ZPB was delivered by infection with a recombinant myxoma virus. In contrast to the results with ZPB protein, infection of rabbits induced a similar serum antibody response to ZPB in both sexes. This indicated that presentation of ZPB in the context of a virus infection was able to overcome tolerance in females. However, the antibody titers were lower than 12 800, and only 25% of female rabbits were infertile. This antibody response was boosted by injections of recombinant ZPB protein, after which 80% of female rabbits were infertile. Infertility was associated with antibody binding to zonae and varying degrees of ovarian pathology characterized by follicular degeneration and substantial depletion of primordial follicles. Oocyte and follicular degeneration appeared to be the principal mechanism of infertility and may be primarily induced by antibodies to ZPB. PMID- 10456836 TI - Sustained but not repeated acute elevation of cortisol impaired the luteinizing hormone surge, estrus, and ovulation in gilts. AB - We tested the hypothesis that sustained and repeated acute elevation of cortisol would impair the LH surge, estrus, and ovulation in gilts. Cortisol was injected intramuscularly, to achieve a sustained elevation of plasma concentrations of cortisol, or intravenously, to achieve an acute elevation of plasma concentrations of cortisol. Control gilts received i.m. injections of oil and i.v. injections of saline. These treatments were administered to gilts (n = 6 per treatment) at 12-h intervals from Days 7 to 11 of the estrous cycle until after estrus ceased or until Day 27 or 28 of the estrous cycle, whichever came first. The repeated acute elevation of cortisol had no effect on the LH surge, estrus, or ovulation. In contrast, when the elevation of cortisol was sustained, the LH surge, estrus, and ovulation were inhibited. We conclude that cortisol is capable of direct actions to impair reproductive processes in female pigs but that plasma concentrations of cortisol need to be elevated for a substantial period for this to occur. PMID- 10456837 TI - Temporal and tissue-specific expression of kallikrein (Klk) genes and identification of a novel Klk messenger ribonucleic acid transcript during early development in the mouse. AB - The kallikreins are a multigene family of serine proteases that act on a diverse number of substrates, including several growth factors and extracellular matrix (ECM) glycoproteins and proteinases. Recently, this family has been implicated in the process of early development and embryo implantation. In this study, we used reverse transcription-polymerase chain reaction with gene-specific primers and Southern hybridization to elucidate the temporal and tissue-specific expression patterns of the mouse kallikreins Klk1, Klk3, Klk5, Klk9, and Klk21 during early development in the embryo, uterus, and decidua. We observed the expression of Klk1 (tissue kallikrein), Klk3 (gamma-nerve growth factor), Klk9 (epidermal growth factor-binding protein), and Klk21 in the early conceptus (until the 2 cell stage). Only Klk21 continued to be expressed in the blastocyst until Day 7.5 of pregnancy. Expression of Klk9 reappeared at Day 7.5 and was consistently detected until Day 11, the last day studied; Klk1 was again expressed in the embryo from Day 9.5, with decreased levels by Day 11. In contrast, in the uterus or decidua, there was no expression of Klk1 until Day 7.5, when mRNA transcripts were abundant; transcripts then decreased in the Day 9.5 and Day 11 uterus. Expression of Klk21 in the uterus and decidua displayed a similar pattern but was detected at much lower levels. Interestingly, a novel Klk21-like mRNA was also detected in uterine tissue samples but not in embryonic samples; Klk3, Klk5, and Klk9 were not consistently expressed in the uterus or decidua over this time. This is the first report of the expression of specific kallikreins during early development. The distinct gene- and tissue-specific expression patterns presented in this study, in conjunction with the well-characterized roles of kallikreins in regulation of protein activation, ECM degradation, and proliferative events, suggests the involvement of the kallikrein gene family during early development. PMID- 10456838 TI - Characterization of the pharmacological-sensitivity profile of neoglycoprotein induced acrosome reaction in mouse spermatozoa. AB - Mammalian spermatozoa undergo the acrosome reaction (AR) in response to the interaction of a carbohydrate-recognizing molecule(s) on the sperm plasma membrane (sperm surface receptor) and its complementary glycan (ligand) moiety(ies) on the zona pellucida (ZP). Previously, we demonstrated that a hexose (mannose) or two amino sugars (glucosaminyl or galactosaminyl residues) when covalently conjugated to a protein backbone (neoglycoproteins) mimicked the mouse ZP3 glycoprotein and induced the AR in capacitated mouse spermatozoa (Loeser and Tulsiani, Biol Reprod 1999; 60:94-101). To elucidate the mechanism underlying sperm-neoglycoprotein interaction and the induction of the AR, we have examined the effect of several AR blockers on neoglycoprotein-induced AR. Our data demonstrate that two known L-type Ca(2+) channel blockers prevented the induction of the AR by three neoglycoproteins (mannose-BSA, N-acetylglucosamine-BSA, and N acetylgalactosamine-BSA). The fact that the L-type Ca(2+) channel blockers (verapamil, diltiazem) had no inhibitory effect on sperm surface galactosyltransferase or alpha-D-mannosidase, two carbohydrate-recognizing enzymes thought to be sperm surface receptors, suggests that the reagents block the AR by a mechanism other than binding to the active site of the enzymes. PMID- 10456839 TI - Autoimmune ovarian inflammation triggered by proinflammatory (Th1) T cells is compatible with normal ovarian function in mice. AB - The detection of noninfectious ovarian inflammation (oophoritis) and serum ovarian autoantibodies in a patient with premature ovarian failure is indicative of an autoimmune etiology. The mechanisms of autoimmune ovarian injury leading to loss of function are currently unknown. In this study we investigated the impact of oophoritis on ovarian function based on two murine autoimmune ovarian disease (AOD) models. AOD can be induced by thymectomy at Day 3 after birth (d3tx). D3tx mice develop ovarian inflammation and atrophy with loss of oocytes. In these mice, ovarian atrophy and not oophoritis correlated with abnormal estrous cyclicity. The second AOD model is induced by active immunization of adult mice with a murine ZP3 peptide (pZP3) in adjuvant. After active immunization, the zona pellucida antibody titer, not oophoritis, correlated with reduced fertility. To investigate the effect of oophoritis in the absence of antibody response or ovarian atrophy, pZP3-specific T cells were passively transferred into naive syngeneic mice. This recruited cytokine-producing cells into the ovaries so that elevated cytokine production and its effect on ovarian function could be examined. Recipients of pZP3-specific T cells developed severe granulomatous oophoritis, and the diseased ovaries had elevated ovarian mRNA levels of interferon-gamma, interleukin-1beta, and tumor necrosis factor alpha. Despite these changes, fertility rates and gonadotropin-induced follicular development remained essentially normal. Therefore, normal ovarian function is compatible with severe ovarian inflammation mediated by autoreactive T cells. PMID- 10456840 TI - Prostaglandin F(2alpha) induces a rapid decline in progesterone production and steroidogenic acute regulatory protein expression in isolated rat corpus luteum without altering messenger ribonucleic acid expression. AB - With interest in steroidogenic acute regulatory protein (StAR) involvement in the luteolytic process, we studied changes in serum progesterone levels and the concomitant expression of StAR mRNA and protein (37-, 32-, and 30-kDa forms) in postovulatory Day 7 corpora lutea (CL) isolated from rats 1 h after injection with prostaglandin F(2alpha) (PGF(2alpha), n = 6) or saline (n = 6). Serum progesterone levels were determined by RIA, StAR and beta-actin mRNA expression by Northern analysis, and StAR and beta-actin protein expression by Western analysis. Adrenal, brain, and spleen from control animals were used as positive and negative controls for StAR expression. Scanning optical densitometry measurements were standardized by dividing the signal strength from each StAR autoradiogram lane by that from the corresponding beta-actin autoradiogram lane. ANOVA was used for significance testing, with alpha set at 0.05. The 37-, 32-, and 30-kDa forms of StAR protein were expressed in all adrenal samples, whereas only the 37- and 30-kDa forms were found in CL. Serum progesterone levels and expression of the 30-kDa and 37-kDa forms of the StAR protein in CL were all found to be significantly lower in the PGF(2alpha)-treated than the saline treated group. StAR mRNA expression was not significantly different in the saline and PGF(2alpha)-treated rats. The rapid decline in StAR protein expression that accompanies PGF(2alpha) induced luteolysis, therefore, does not result from significant decline in mRNA expression. PMID- 10456841 TI - Inhalation of mainstream and sidestream cigarette smoke retards embryo transport and slows muscle contraction in oviducts of hamsters (Mesocricetus auratus). AB - Prior experiments have shown that the functioning of hamster oviducts is impaired by in vitro exposure to cigarette smoke. To determine if cigarette smoke affects oviductal functioning in vivo, an inhalation experiment was done in which hamsters were exposed to doses of smoke similar to those received by human smokers. The effects of mainstream smoke (the bolus of smoke inhaled by active smokers) and sidestream smoke (the main component in environmental tobacco smoke) were compared. Transport of preimplantation embryos through the hamster oviduct was retarded in females inhaling doses of mainstream or sidestream smoke that produced serum cotinine levels within the range reported for women who actively or passively smoke during pregnancy. In addition, hamster oviductal muscle contraction rate decreased significantly during a single exposure of animals to either mainstream or sidestream smoke, and contraction rate failed to return to initial control values during a 25-min recovery period. Both preimplantation embryo transport and muscle contraction were more sensitive to sidestream than mainstream smoke. These data demonstrate that inhalation of doses of mainstream and sidestream cigarette similar to those received by active and passive human smokers adversely affects functioning of the oviduct and may explain the increased incidence of ectopic pregnancies reported in women who smoke. PMID- 10456842 TI - Effect of inflammatory mediators on the electrophysiology of the human oviduct. AB - The effects of histamine and other inflammatory mediators on the electrophysiology and intracellular free calcium ([Ca(2+)](i)) of human oviductal epithelial cells, grown as a polarized layer in primary culture, were studied. Transepithelial potential difference (PD) and short-circuit current (I(scc)) were recorded using a modified Ussing chamber. Resistance (R) was calculated from the measurements of PD and I(scc). Basally applied histamine produced transient increases in PD and I(scc) with a small decrease in R. The histamine effect was reduced by triprolidine (H(1) receptor antagonist) but was unaffected by H(2) (ranitidine) or H(3) (thioperamide) receptor antagonists. Blockers of Na(+), K(+), or Na(+)/K(+)/2Cl(-) channels did not affect histamine action. Blockers of Cl(-)/HCO(3)(-) channels or Ca(2+) channels reduced the histamine effect. Platelet activating factor (PAF), applied apically, increased PD and I(scc). Histamine produced a transient increase in fluorescence of Fura 2-AM dye, indicating an increase in [Ca(2+)](i). Triprolidine pretreatment inhibited histamine-stimulated [Ca(2+)](i) increase. Cimetidine, (H(2) receptor antagonist), ranitidine, or thioperamide reduced the histamine effect. Histamine increased contractions of both circular and longitudinal smooth muscles in oviduct segments, an effect that was antagonized by triprolidine or thioperamide but not by ranitidine. Histamine's action on Ca(2+) and Cl(-) movements may adversely affect oviductal fluid production and decrease fertility. PAF's effects on Cl(-) movements may be important for normal embryo transport. PMID- 10456843 TI - Apolipoprotein E peptide stimulation of rat ovarian theca cell androgen synthesis is mediated by members of the low density lipoprotein receptor superfamily. AB - Ovarian androgen production is rate limiting for follicular maturation and can induce follicular atresia. Thus, it is important to define the actions of the intraovarian agents, such as apolipoprotein (apo) E, that modulate theca cell androgen production. Theca cell androgen production is stimulated at low concentrations and inhibited at higher concentrations of native apo E. The apo E peptide, acetyl-Y(LRKLRKRLLRDADDL)(2)C or acetyl-Y(141-155)(2)C, has low density lipoprotein (LDL) receptor and LDL receptor-related protein-binding activity, and it mimics the activity of native apo E in the theca-interstitial cell system. To define the role of members of the LDL receptor superfamily in the apo E peptide mediated responses, we found that receptor-associated protein prevented the stimulation without altering the inhibition of androstenedione production. The apo E peptide (129-162), which has no LDL receptor-binding activity, did not stimulate androstenedione production. The apo E peptide acetyl-Y(141-155)(2)C did not stimulate androstenedione production when cell surface heparan sulfate proteoglycans were degraded with heparinase. The apo E peptide acetyl-Y(141 155)(2)C bound to heparin, a property of LDL receptor ligands, and in this complex the peptide had no effect on androstenedione production. These observations support the conclusion that apo E-mediated stimulation, but not inhibition, of ovarian theca cell androstenedione production was mediated by members of the LDL receptor superfamily. PMID- 10456844 TI - Strontium supports human sperm capacitation but not follicular fluid-induced acrosome reaction. AB - The ability of strontium (Sr(2+)) to replace calcium (Ca(2+)) in maintaining human sperm function has still not been completely characterized. In the present study, acrosome reaction (AR) inducibility in response to human follicular fluid (hFF) was compared in spermatozoa incubated in either Ca(2+)- or Sr(2+) containing media. Other events related to sperm capacitation, such as protein tyrosine phosphorylation and hyperactivation as well as zona pellucida (ZP) recognition under both conditions, were also analyzed. Spermatozoa incubated overnight in the presence of Sr(2+) were unable to undergo the AR when exposed to hFF. Nevertheless, when spermatozoa were incubated under this condition and then transferred to medium with Ca(2+), sperm response to hFF was similar to that of cells incubated throughout in the presence of Ca(2+). The sperm protein tyrosine phosphorylation patterns and the percentages of sperm motility and hyperactivation were similar after incubation in Ca(2+)- or Sr(2+)-containing media. Under both conditions, the same binding capacity to homologous ZP was observed. Similar results were obtained when EGTA was added in order to chelate traces of Ca(2+) present in Sr(2+) medium. From these results, it can be concluded that Sr(2+) can replace Ca(2+) in supporting capacitation-related events and ZP binding, but not hFF-induced AR of human spermatozoa. PMID- 10456845 TI - Identification of a stress-induced protein during human trophoblast differentiation by differential display analysis. AB - Differentiation of human placental trophoblast is characterized by a process during which mononuclear villous cytotrophoblasts fuse to form a multinucleate syncytium. This event is associated with dramatic changes in gene expression. In the present study, we have applied a sensitive approach-differential display analysis-to evaluate changes in gene expression during in vitro forskolin-induced differentiation of a model of human trophoblast, the choriocarcinoma BeWo. We identified seven genes that were up-regulated; their expression and function have not previously been reported in trophoblast. Four up-regulated genes were novel upon comparison of their sequences to the GenBank database. The other three genes encode human cytochrome p450 IIC, inosine monophosphate dehydrogenase type II, and reducing agent and tunicamycin-responsive protein (RTP). Northern blot analysis revealed that RTP mRNA expression was induced to 3-fold in BeWo after 24 h incubation with forskolin and increased up to 11-fold by 72 h of forskolin treatment. The expression pattern of RTP was further investigated by in situ hybridization on second trimester and term placenta tissues. RTP mRNA was predominantly expressed in syncytiotrophoblasts in both second trimester and term placentae. The expression of RTP gene in BeWo cells was protein kinase C dependent. This is the first description of RTP gene expression in placenta and the first study elucidating the signaling pathway involved in the regulation of RTP gene expression. These results suggest that RTP may play a role in trophoblast cell proliferation and differentiation. PMID- 10456846 TI - Glyceraldehyde-3-phosphate dehydrogenase expression during apoptosis and proliferation of rat ventral prostate. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional enzyme known to play a critical role in neuronal apoptosis. We undertook the current studies to determine whether GAPDH also plays a role in prostate epithelial cell apoptosis in response to androgen deprivation. To do so, we analyzed GAPDH staining by immunohistochemistry during castration-induced involution and androgen-induced regeneration of rat ventral prostate. We found that GAPDH was undetectable in secretory epithelial cells at baseline and that staining did not increase in the epithelium during the period of peak apoptosis from 1 to 3 days after castration. However, GAPDH levels did increase within nuclei of some basal epithelial cells 5 days after castration and within the cytoplasm of all secretory epithelial cells 7 days after castration. GAPDH was also abundant within the cytoplasm of secretory epithelial cells during the period of maximal cell proliferation from 2 to 3 days after androgen replacement and was clearly apparent within nuclei of some epithelial cells 4 days after androgen replacement. Our studies suggest that GAPDH plays multiple roles during prostate epithelial cell apoptosis and proliferation. PMID- 10456847 TI - Cadherin-catenin complexes during zebrafish oogenesis: heterotypic junctions between oocytes and follicle cells. AB - During vertebrate oogenesis, the germ cells and associated somatic cells remain connected by a variety of adhering junctional complexes. However, the molecular composition of these cellular structures is largely unknown. To identify the proteins forming the heterotypic adherens junctions between oocytes and follicle cells in the zebrafish (Danio rerio), the cDNAs encoding alphaE-catenin and plakoglobin were isolated. Using these cDNAs, in combination with the previously isolated beta-catenin cDNA, and antibodies specific for alpha- and beta-catenin, plakoglobin, and N- and E-cadherin, we found differences in catenin and plakoglobin gene expression during oogenesis. The immunolocalization of these plaque proteins, as well as of cadherins, in the ovarian follicle indicated an enrichment of alpha- and beta-catenin and of E-cadherin-like protein(s) in the oocyte cortex, notably at sites of oocyte-follicle cell contacts, suggesting the presence of hitherto unknown heterotypic adherens junctions between these cells. By contrast, plakoglobin and N-cadherin localization was restricted to cell-cell contacts in the follicle cell layer. During oocyte maturation, mRNAs for alphaE- and beta-catenin and plakoglobin accumulated, and all three plaque-forming proteins were stored in unfertilized eggs, either in complexed forms with cadherins or as free cytoplasmic pools. These findings suggest possible roles of these junctional proteins during early embryogenesis. PMID- 10456848 TI - Distribution of immune cells in the epididymis of the aging Brown Norway rat is segment-specific and related to the luminal content. AB - Remarkable changes occur during aging in the testis and epididymis of the Brown Norway rat. A dramatic increase in the number of halo cells, which are present in the epididymal epithelium and originate from the immune system, is found in animals of increasing age. Halo cells have been postulated to be either lymphocytes or monocytes. We hypothesized that halo cells are a mixture of different immune cells and that their relative composition changes with age. To verify this hypothesis, markers for helper T lymphocytes, cytotoxic T lymphocytes, B lymphocytes, and monocytes-macrophages were used to identify the major categories of immune cells in the epididymides of Brown Norway rats ranging in age from 3 to 24 mo. The numbers of immunocompetent cells in the epididymis were determined in relation to age, epididymal segment, and luminal content. We found that monocytes, helper T lymphocytes, and cytotoxic T lymphocytes belong to the population of halo cells. In addition, a segment-specific increase with age in the number of these immune cells was noted. Finally, we report a segment specific recruitment of cytotoxic T lymphocytes and monocytes-macrophages in the epididymal epithelium of aged rats whose epididymal lumen contained few spermatozoa. We postulate that accumulation of damaged epithelial cells and antigens of germ cell origin, leaking through a dysfunctional blood-epididymis barrier, may contribute to the active recruitment of immune cells with age. PMID- 10456849 TI - Gonadotropin-releasing hormone activates the equine luteinizing hormone beta promoter through a protein kinase C/mitogen-activated protein kinase pathway. AB - GnRH regulation of LH secretion is well understood and involves Ca(2+) mobilization. However, the mechanism by which GnRH activates transcription of the LHbeta gene is controversial. GnRH is known to elevate intracellular calcium and activate the protein kinase C (PKC) pathway. The present study evaluated the pathway(s) involved in GnRH induction of LHbeta transcription. We have previously reported that the equine LHbeta (eLHbeta -448/+60) promoter is active in alphaT3 1 cells. Therefore, we created a clonal, stably transfected alphaT3-1 gonadotroph cell line harboring the eLHbeta promoter (-448/+60) fused to the luciferase reporter gene. Administration of a GnRH agonist resulted in induction of promoter activity that was completely inhibited by the antagonist antide. Various calcium affecting drugs had no effect on the promoter. Administration of phorbol 12 myristate 13-acetate (PMA) elicited an activation similar to, albeit lower than, that with GnRH. Down-regulation or pharmacological inhibition of PKC completely blocked PMA's induction of the promoter, while GnRH induction was only partly attenuated. Treatment with the mitogen-activated protein kinase (MAPK) kinase inhibitor, PD98059, completely inhibited the activation of eLHbeta by PMA but only partly diminished GnRH's induction. Expression of the transcription factor, early growth response protein 1 (Egr1), correlated completely with activation of MAPK, suggesting that Egr1 is the factor through which PKC/MAPK acts. Our data suggest that GnRH induces activity of the eLHbeta promoter by activating a signal transduction cascade involving PKC-MAPK-Egr1 but that has no significant requirement for calcium. PMID- 10456850 TI - Experimental assumption of dominance by a smaller follicle and associated hormonal changes in mares. AB - A two-follicle model was used to study the nature of selection of the dominant follicle in mares by ablating neither or one of the two follicles on the day the larger follicle reached >/= 20 mm (Day 0). The larger follicle became the dominant follicle in all mares in which both follicles (n = 8) or only the larger follicle (n = 10) was retained. When only the smaller follicle (n = 9) was retained, it became dominant and ovulated in six mares and became atretic in three mares; the difference in diameter between the two follicles on Day 0 was less (p < 0.01) in mares in which the retained smaller follicle grew and ovulated (2.2 +/- 0.6 mm) than in the mares in which the follicle became atretic (5.9 +/- 1.2 mm). A decline (p < 0. 0001) in FSH concentrations occurred over Days -4 (8.4 +/- 0.7 ng/ml) to 0 (5.9 +/- 0.3 ng/ml), averaged over all groups, and the decline continued for several more days in the groups with both follicles or with only the larger follicle retained. In the group with only the smaller follicle retained, compared to the group with both follicles retained, FSH concentrations and diameter of the smaller follicle increased between Days 0 and 1 (significant interaction for each end point). After Day 1, FSH concentrations continued to increase when the smaller retained follicle became atretic; concentrations decreased when the smaller retained follicle became dominant. An increase (p < 0.0001) in LH concentrations occurred over Days -4 (12.2 +/- 1.1 pg/ml) to 0 (21.1 +/- 2.0 pg/ml), averaged over the three groups. In 23 of 27 mares, a transient peak in LH concentrations occurred within 2 days of Day 0. In the groups with both follicles or with only the larger follicle retained, an increase (p < 0.0001) in systemic estradiol concentrations occurred between Day 0 (5.3 +/- 0.6 pg/ml) and Day 2 (7.5 +/- 0.4 pg/ml). When only the smaller follicle was retained, estradiol did not begin to increase until Day 2, and it increased only when the retained follicle grew and became dominant. The beginning of an increase in estradiol and continued decrease in FSH at the expected beginning of deviation were attributable to the future dominant follicle; there was no indication that the smaller follicle was involved. PMID- 10456851 TI - Temporal and differential effects of amino acids on bovine embryo development in culture. AB - The aim of the study was to determine the amino acid requirements of the in vitro produced bovine embryo as it develops from the zygote to the blastocyst, using a two-step culture system. When added to synthetic oviduct fluid (SOF) for the first 72-h culture, Eagle's nonessential amino acids and glutamine (NeGln) significantly increased development to the 8- to 16-cell stage (Day 4 postinsemination [pi]) and subsequent blastocyst development (Day 7 pi). Glutamine alone during the first 72-h culture did not stimulate development to the 8- to 16-cell stage (p > 0.05); however, the removal of glutamine from NeGln reduced the stimulatory effects of the nonessential amino acids. Replacing glutamine with betaine (an organic osmolyte) in NeGln did not stimulate development to the 8- to 16-cell stage compared to culture in SOF, but it did improve subsequent blastocyst development, indicating an osmolytic function of glutamine during the first 72-h culture. The addition of Eagle's essential amino acids and glutamine to SOF, or to medium already containing nonessential amino acids and glutamine for the first 72-h culture, did not affect cleavage to the 8- to 16-cell stage or subsequent blastocyst development (p > 0.05). Beyond Day 4 pi, culture with 20aa (nonessential and essential amino acids and glutamine) increased blastocyst development, total cell number, and the number of cells in both the trophectoderm and inner cell mass, compared to culture with other groups of amino acids (p < 0.05). Substituting betaine for glutamine in 20aa reduced blastocyst formation, indicating a non-osmolytic function of glutamine during the second 72-h culture. Further, there was a significant negative correlation between the concentration of essential amino acids (quarter, half, or single strength) and embryo development during both the first 72-h and second 72-h culture (p < 0.01), indicating that the concentration of essential amino acids was too high during culture of the bovine embryo. This study identified the temporal and differential effects of amino acids during development of the bovine embryo from the zygote to the blastocyst. PMID- 10456853 TI - Role of egg sulfolipidimmobilizing protein 1 on mouse sperm-egg plasma membrane binding. AB - We have shown that sperm sulfolipidimmobilizing protein 1 (SLIP1, molecular mass of 68 kDa), a sulfogalactosylglycerolipid (SGG)-binding protein, is significant in sperm-zona pellucida (ZP) interaction. The objective of this study was to localize SLIP1 on the egg and determine its role in gamete interaction. Immunofluorescence and immunoprotein A gold electron microscopy localized SLIP1 to the egg plasma membrane. In vitro gamete binding, using zona-free eggs preincubated with antiSLIP1 Fab before coincubation with sperm, showed a significant, dose-dependent decrease in sperm-egg plasma membrane binding. Similar results were obtained when affinity-purified antiSLIP1 IgG was used for egg pretreatment. The significance of egg SLIP1 in sperm-egg plasma membrane binding was further demonstrated by a decrease (36-52%) in in vitro fertilization when zona-intact eggs were pretreated with antiSLIP1 IgG. Since SLIP1 has been shown to bind SGG in vitro, we investigated the possibility that sperm SGG may participate in sperm-egg plasma membrane binding through egg SLIP1. Pretreatment of sperm with antiSGG Fab prior to coincubation with zona-free eggs resulted in a dose-dependent decrease in sperm-egg plasma membrane binding. Collectively, these findings strongly suggest a role for egg SLIP1 in sperm-egg plasma membrane interaction, which may be through its binding to sperm SGG. PMID- 10456852 TI - Reverse transcription-polymerase chain reaction analysis of genes involved in gonadal differentiation in pigs. AB - In mammals, testis development is initiated in the embryo as a response to the expression of the sex-determining gene, SRY. The time course of SRY expression during gonadal differentiation in the male has been described in detail only in mice and sheep. In this study, we used reverse transcription-polymerase chain reaction analysis to define the SRY transcription profile in pig genital ridges. SRY transcripts were first detectable from 23 days postcoitum (dpc), then declined sharply after 35 dpc. None were detected at 60 dpc. In addition, we analyzed temporal expression of other genes known to be involved in mammalian sex determination: WT-1, SF-1, SOX9, and AMH. A key stage seems to be 28 dpc, in which SOX9 expression switches between the male and female, and AMH expression begins to attest to Sertoli cell differentiation and to correspond to seminiferous cord formation in the male. Expression of gonadotropin receptors and aromatase was also investigated in porcine gonads, and we showed that their transcripts were detected very early on, especially in the male: 25 dpc for the LH receptor (rLH) and aromatase, and 28 dpc for the FSH receptor (rFSH). In the female, aromatase transcripts were not detected until 70 dpc, and rFSH expression occurred later: at 45 dpc at the onset of meiosis. Moreover, no difference was observed between the sexes for the onset of rLH transcription at 25 dpc. Such a thorough study has never been performed on pigs; developmental analysis will be useful for investigating sex-reversed gonads and determining ontogeny in intersexuality, a common pathology in pigs. PMID- 10456854 TI - Quantification of prolactin messenger ribonucleic acid, pituitary content and plasma levels of prolactin, and detection of immunoreactive isoforms of prolactin in pituitaries from turkey embryos during ontogeny. AB - The content of prolactin mRNA as well as total prolactin content and type of isoforms of prolactin were measured in single pituitary glands from turkey embryos and poults. Levels of mRNA and pituitary content of prolactin remained low until 5 days before hatching, while plasma concentrations remained low until 2 days before hatching. Levels of prolactin mRNA then increased until the day of hatch, stayed stable during the 3 first days of age, and significantly increased until 2 wk of age. Similar changes were observed in pituitary content and plasma levels of prolactin. Two immunoreactive bands of apparent molecular masses of 24 and 27 kDa, corresponding to the nonglycosylated and glycosylated form of prolactin, respectively, were visualized on Western blots. In pituitary glands from embryos at 22 days of incubation, 31.5% of the protein was glycosylated, whereas in embryos at 27 days of incubation and poults at 1 and 7 days of age, 48.6%, 48.0%, and 56. 0% of prolactin was glycosylated, respectively. The results indicate that the increases in the synthesis and the release of prolactin occur mainly around and after the time of hatching in the turkey embryo. Higher percentages of glycosylated isoforms were associated with increasing levels of total prolactin in the pituitary gland. Thus, the synthesis of prolactin and its post-translational modifications may be important factors involved in the physiologic changes occurring around the time of hatching. PMID- 10456855 TI - Subzero water permeability parameters of mouse spermatozoa in the presence of extracellular ice and cryoprotective agents. AB - Optimization of techniques for cryopreservation of mammalian sperm is limited by a lack of knowledge regarding water permeability characteristics during freezing in the presence of extracellular ice and cryoprotective agents (CPAs). Cryomicroscopy cannot be used to measure dehydration during freezing in mammalian sperm because they are highly nonspherical and their small dimensions are at the limits of light microscopic resolution. Using a new shape-independent differential scanning calorimeter (DSC) technique, volumetric shrinkage during freezing of ICR mouse epididymal sperm cell suspensions was obtained at cooling rates of 5 and 20 degrees C/min in the presence of extracellular ice and CPAs. Using previously published data, the mouse sperm cell was modeled as a cylinder (122-microm long, radius 0.46 microm) with an osmotically inactive cell volume (V(b)) of 0.61V(o), where V(o) is the isotonic cell volume. By fitting a model of water transport to the experimentally obtained volumetric shrinkage data, the best-fit membrane permeability parameters (L(pg) and E(Lp)) were determined. The "combined best-fit" membrane permeability parameters at 5 and 20 degrees C/min for mouse sperm cells in solution are as follows: in D-PBS: L(pg) = 1.7 x 10(-15) m(3)/Ns (0.01 microm/min-atm) and E(Lp) = 94.1 kJ/mole (22.5 kcal/mole) (R(2) = 0.94); in "low" CPA media (consisting of 1% glycerol, 6% raffinose, and 15% egg yolk in D-PBS): L(pg)[cpa] = 1.7 x 10(-15) m(3)/Ns (0.01 microm/min-atm) and E(Lp)[cpa] = 122.2 kJ/mole (29.2 kcal/mole) (R(2) = 0.98); and in "high" CPA media (consisting of 4% glycerol, 16% raffinose, and 15% egg yolk in D-PBS): L(pg)[cpa] = 0.68 x 10(-15) m(3)/Ns (0.004 microm/min-atm) and E(Lp)[cpa] = 63.6 kJ/mole (15.2 kcal/mole) (R(2) = 0.99). These parameters are significantly different than previously published parameters for mammalian sperm obtained at suprazero temperatures and at subzero temperatures in the absence of extracellular ice. The parameters obtained in this study also suggest that damaging intracellular ice formation (IIF) could occur in mouse sperm cells at cooling rates as low as 25-45 degrees C/min, depending on the concentrations of the CPAs. This may help to explain the discrepancy between the empirically determined optimal cryopreservation cooling rates, 10-40 degrees C/min, and the numerically predicted optimal cooling rates, greater than 5000 degrees C/min, obtained using suprazero mouse sperm permeability parameters that do not account for the presence of extracellular ice. As an independent test of this prediction, the percentages of viable and motile sperm cells were obtained after freezing at two different cooling rates ("slow" or 5 degrees C/min; "fast," or 20 degrees C/min) in both the low and high CPA media. The greatest sperm motility and viability was found with the low CPA media under fast (20 degrees C/min) cooling conditions. PMID- 10456856 TI - Follicle-stimulating hormone receptor and its messenger ribonucleic acid are present in the bovine cervix and can regulate cervical prostanoid synthesis. AB - The hypothesis that FSH regulates the bovine cervical prostaglandin E(2) (PGE(2)) synthesis that is known to be associated with cervical relaxation and opening at the time of estrus was investigated. Cervical tissue from pre-estrous/estrous, luteal, and postovulatory cows were examined for 1) the presence of bovine (b) FSH receptor (R) and its corresponding mRNA and 2) the effect of FSH on the PGE(2) regulatory pathway in vitro. The presence of bFSHR mRNA in the cervix (maximal during pre-estrus/estrus) was demonstrated by the expression of a reverse transcription (RT) polymerase chain reaction (PCR) product (384 base pairs) specific for bFSHR mRNA and sequencing. Northern blotting revealed three transcripts (2.5, 3.3, and 3.8 kilobases [kb]) in cervix from pre-estrous/estrous cows. The level of FSHR (75 kDa) was significantly higher (p < 0.01) in Western blots of pre-estrous/estrous cervix than in other cervical tissues. There was a good correlation between the 75-kDa protein expression and its corresponding transcript of 2.55 kb throughout the estrous cycle as described by Northern blot analysis as well as RT-PCR. Incubation of FSH (10 ng/ml) with pre-estrous/estrous cervix resulted in a 3-fold increase in the expression of FSHR and a 2-fold increase in both G protein (alpha(s)) and cyclooxygenase II. FSH (5-20 ng/ml) significantly increased (p < 0.01) cAMP, inositol phosphate (p < 0.01), and PGE(2) (p < 0.01) production by pre-estrous/estrous cervix but not by cervix at the other stages. We conclude that bovine cervix at the time of the peripheral plasma FSH peak (pre-estrus/estrus) contains high levels of FSHR and responds to FSH by increasing the PGE(2) production responsible for cervical relaxation at estrus. PMID- 10456857 TI - Molecular cloning and expression of ovarian cathepsin D in seabream, Sparus aurata. AB - In marine fish producing pelagic eggs, the acquirement of buoyancy by the eggs through the hydration process is a key event of reproduction; moreover, the yolk proteolysis, which leads to buoyancy, seems to affect the fertility and survival of the spawned eggs. Recently we demonstrated that cathepsin D is the aspartic protease responsible for this intraoocytic processing of vitellogenin into yolk proteins. In the present study, we isolated, cloned, and sequenced the cDNA encoding cathepsin D and studied expression of the message by Northern blotting and whole-mount in situ hybridization. The full-length seabream cathepsin D cDNA is 1837 base pairs long, encoding a protein of 400 amino acids (aa) consisting of a signal peptide of 19 aa, a prosequence of 44 aa, and a mature peptide of 336 aa. An absolute sequence conservation at the aspartyl residues (+33 and +221) was found, and there are three potential N-glycosylation sites at +70, aa +189, and aa +274. The aa sequence of seabream cathepsin D reveals a high degree of sequence similarity with cathepsin D mRNAs from other organisms (73% sequence homology to mouse and rat, 72% to human and trout, 69% to chicken, 66% to pig, and 65% to Xenopus). The cathepsin D mRNA in floating eggs was present as a single band that was approximately 1.9 kilobases (kb) in size, while in the sinking eggs there were several fast-migrating bands (size range 1.3-0.2 kb). Whole-mount hybridization was used to investigate transcription of cathepsin D in the developing embryo; during the hatching period, cathepsin D mRNA-positive cells were distributed in a wide region between the trunk and the tail, and in the ventral region over the yolk ball. The highest levels of cathepsin D enzymatic activity were found in the sinking eggs and during the hatching period of embryonic development. These data suggest that cathepsin D can be considered a possible marker for egg quality. PMID- 10456858 TI - Contribution of branched-chain amino acids to uteroplacental ammonia production in sheep. AB - The uteroplacental tissues are a principal site of ammonia production for the conceptus. The goal of this study was to examine the effect of the composition of maternal amino acid (AA) infusate on uteroplacental ammonia production. Seven pregnant ewes (126 +/- 1. 4 days gestation) were infused through the maternal femoral vein (duration 3.5 h, rate 240 ml per hour) with three solutions of AAs. The first infusate was comparable to commercial parenteral nutrition preparations, the second infusate contained the same solution without branched chain AAs (BCAAs), and the third infusate contained only BCAAs. Blood samples were simultaneously collected from the maternal artery, uterine vein, fetal artery, and umbilical vein to determine plasma AA concentrations and whole blood ammonia concentrations, before (control) and 2 h after (experimental) the start of infusion. Uterine and umbilical blood flows were measured using the ethanol steady-state diffusion method. Results showed that fetal arterial and venous ammonia concentrations increased significantly after infusions with all AAs or only BCAAs, but not without BCAAs. Uteroplacental ammonia production increased in response to each of the three infusates. However, this increase was much greater when the BCAAs were present in infusates. We conclude that there is a significant contribution of BCAAs to the uteroplacental ammonia production. Maternal AA infusions containing BCAAs can result in increased fetal blood ammonia concentrations. PMID- 10456859 TI - Immunolocalization of progesterone receptors in bovine placentomes throughout mid and late gestation and at parturition. AB - The corpus luteum is the main source of progesterone (P(4)) responsible for maintenance of gestation in cattle. So far it has not been possible to assign any biological role to placental P(4), which contributes only marginally and temporarily to peripheral maternal blood levels. In order to identify possible P(4) target cells within the placenta, placentomes from 150-, 220-, 240-, and 270 day-pregnant cows and from parturient cows (3 animals per group) were screened immunohistochemically for expression of the progesterone receptor (PR). During gestation, PR-positive staining was found exclusively in the nuclei of caruncular stromal cells (CSC; maternal part of the placentome) and of caruncular vascular pericytes. In placentomes from parturient cows, occasional positive nuclear staining was also observed in the walls of small caruncular arteries. The percentage of PR-positive CSC increased slightly from 51.8 +/- 2.6% on Day 150 to 56.2 +/- 5.6% at Day 270 (p < 0.05) and was 58.9 +/- 1.8% at parturition. These results suggest that in pregnant cattle, CSC are under the control of P(4) of placental rather than luteal origin. Thus, whereas luteal P(4) may regulate "coarse" systemic progestational functions in the maternal compartment in the classical hormonal manner, placental P(4) may act as a paracrine factor involved in the local regulation of caruncular growth, differentiation, and functions. PMID- 10456861 TI - Chromosome analysis of BALB/c mouse spermatozoa with normal and abnormal head morphology. AB - To assess the relationship between mouse sperm head morphology and karyotype, sperm heads with either a normal or an abnormal morphology were injected individually into enucleated mouse oocytes that were karyotyped at the metaphase of the first cleavage. BALB/c male mice that produce an unusually high proportion of morphologically abnormal spermatozoa were used as sperm donors. Abnormal karyotypes were found in a significantly higher proportion of eggs injected with severely misshapen sperm heads (36-38%) as compared to those injected with normal and quasi-normal heads (15-21%) (p < 0.01). Most karyotype abnormalities were structural rather than numerical, the most common being breaks and exchanges of chromosome type in both normal and abnormal spermatozoa. PMID- 10456860 TI - CD26 and adenosine deaminase interaction: its role in the fusion between horse membrane vesicles and spermatozoa. AB - Membrane vesicles of horse seminal plasma present at their surface a highly specific serine-type protease, dipeptidyl peptidase IV/CD26, a surface antigen known to characterize human prostasomes. Horse sperm cells expressed at their surface A(1) adenosine receptors (A(1)AR) and ecto-adenosine deaminase (ecto ADA), both detected by immunoblot analysis, whereas CD26 was visualized at the equatorial segment by immunofluorescence microscopy. In addition to CD26, horse membrane vesicles showed ecto-ADA. The fusion process between horse sperm cells and vesicles was evidenced by confocal microscopy, which showed the localization of CD26 at the postacrosomal region and at the midpiece of the spermatozoa after incubation with vesicles. Moreover, a similar localization of CD26 and ecto-ADA on the spermatozoa was evidenced after fusion. Our results suggest that the interaction CD26/ecto-ADA might be responsible for fusion. Since A(1)ARs are said to be second receptors for ecto-ADA to form ecto-ADA/A(1)AR complexes, and since horse spermatozoa have A(1)ARs at their surface, the interaction CD26/ecto ADA/A(1)AR during the fusion process cannot be ruled out. PMID- 10456862 TI - Completion of meiosis is not always required for acrosome formation in HSP70-2 null mice. AB - Hsp70-2 is a unique member of the mouse 70-kDa heat shock protein family that is synthesized during meiosis in spermatogenic cells. Germ cells in male mice homozygous for a targeted mutation in the Hsp70-2 gene (Hsp70-2(-/-)) arrest in development and undergo apoptosis at the end of the pachytene spermatocyte stage of meiotic prophase. However, cells with a putative acrosome were present occasionally in histological sections of the testes of juvenile and adult Hsp70 2(-/-) mice. This study verified that acrosomes were present and investigated the relationship between acrosome formation and the process of meiosis. Histochemistry with the periodic acid-Schiff procedure and immunostaining with monoclonal antibody MN7 verified that acrosomes were present in Hsp70-2(-/-) mice, and electron microscopy showed that some of these cells had condensing nuclei characteristic of step 8-9 spermatids. The frequency of acrosome containing cells in Hsp70-2(-/-) mice was less than 0.01% of that in wild-type mice. Propidium iodide staining and cytophotometry indicated that the average DNA content of nuclei in MN7-positive cells in Hsp70-2(-/-) mice was usually about twice, or occasionally the same as, that of nuclei in round spermatids of wild type mice. Meiotic metaphase I and II chromosome spreads were observed in spermatogenic cells from Hsp70-2(-/-) mice but at a much lower frequency than in wild-type mice. These results indicate that not all pachytene spermatocytes in Hsp70-2(-/-) mice arrest in meiosis, but they may divide once or sometimes twice and begin acrosome formation and nuclear condensation. This demonstrates that some aspects of spermatid development can occur without the completion of meiosis in mice, as has been reported recently for Drosophila. PMID- 10456863 TI - Interleukin-8 release from human gestational tissue explants: effects of gestation, labor, and chorioamnionitis. AB - Interleukin-8 (IL-8) is a chemotactic cytokine that has been implicated in the process of human parturition, including the processes of cervical ripening and rupture of fetal membranes. In this study, the in vitro release of IL-8 from human amnion, choriodecidua, and placenta tissues obtained before and after spontaneous labor onset both at term and preterm, was assessed. The effect of chorioamnionitis on IL-8 release was also established. All tissue explants examined released IL-8; however, IL-8 release from choriodecidual explants was significantly (p < 0.02) greater than that observed from amnion and placenta. Furthermore, choriodecidual IL-8 release was significantly (p < 0.001) greater from term tissues (850 +/- 134.4 ng/mg DNA, n = 18) than from preterm tissues (458 +/- 68.8 ng/mg DNA, n = 17). Spontaneous onset of labor, irrespective of the eventual mode of delivery, was not associated with any significant changes in IL 8 release from human gestational tissues compared to not-in-labor tissues, both at term and preterm. IL-8 release from gestational tissues was not significantly different in the absence or presence of chorioamnionitis. These data are in contrast to the previously reported stimulatory effects of bacterial endotoxin on IL-8 release from human gestational tissues. The data are consistent, however, with the suggestion that IL-8 release is an early event in chorioamnionitis that precedes the appearance of clinically overt symptoms. PMID- 10456864 TI - Male pronuclear formation in denuded porcine oocytes after in vitro maturation in the presence of cysteamine. AB - The present study was conducted to examine effects of cysteamine in culture medium on progression of meiosis, glutathione (GSH) content, kinase activities (histone H1 kinase and mitogen-activated protein kinase), and male pronuclear formation after in vitro insemination of cumulus-denuded oocytes (DOs) in the pig. DOs, obtained by mechanically removing cells from cumulus-oocyte complexes (COCs) with a small-bore pipette, were cultured for 45 h in TCM199 supplemented with sodium pyruvate, gonadotropins, estradiol, and 10% porcine follicular fluid, with or without cysteamine (150 microM). Maturation rates of DOs cultured with and without cysteamine were not different (60-70%) but were significantly lower than those of COCs (90-100%) (p < 0.05). GSH content of matured DOs cultured with cysteamine was significantly higher than that of DOs cultured without cysteamine (p < 0.05). Values for both types of kinase activity in matured DOs cultured with and without cysteamine were not different (p > 0.05). After in vitro insemination, DOs cultured with cysteamine showed significantly higher rates of male pronuclear formation (80.3 +/- 3.0%) than DOs cultured without cysteamine (16.4 +/- 0.5%) (p < 0.05). These results indicate that the addition of cysteamine to culture medium increased oocyte GSH content and promoted male pronuclear formation after sperm penetration of porcine DOs but had no effects on their maturation rates or kinase activities. PMID- 10456865 TI - Gonadal stage-dependent effects of gonadal steroids on gonadotropin II secretion in the Atlantic croaker (Micropogonias undulatus). AB - Involvement of gonadal steroids in the control of gonadotropin II (GTH II) (homologous to LH) secretion was investigated in the Atlantic croaker (Micropogonias undulatus) using gonadectomy (Gx) and steroid replacement paradigms. Gonadectomy in males and females during the late gonadal recrudescence phase elicited significant increases in the gonadotropin response to stimulation by an LHRH analog (LHRHa), without altering basal GTH II secretion. Slow-release silicone elastomer implants of testosterone or estradiol significantly inhibited LHRHa-induced GTH II secretion in gonad-intact and Gx males, and in Gx females, whereas 5alpha-dihydrotestosterone, a nonaromatizable androgen, was ineffective. Pretreatment of fish with an aromatase inhibitor, 1,4, 6-androstatrien-3,17 dione, 2 days before the administration of testosterone implants, completely blocked the negative effect of testosterone on LHRHa-induced GTH II secretion in males, but only partially restored it in females. This suggests that the negative feedback of testosterone in males is primarily mediated by its conversion to estradiol at the level of the hypothalamus and/or pituitary gland, while in females the androgen may also exert a direct inhibitory effect on GTH II secretion, probably mediated via an androgen receptor. In addition, estradiol and testosterone exerted positive effects on basal and LHRHa-induced GTH II secretion during the early-recrudescence phase of the gonadal cycle. The steroids switched to a negative effect on LHRHa-induced GTH II secretion once the fish had fully developed gonads, possibly as a mechanism that prevents a precocious surge in GTH II secretion and final gamete maturation until gametogenesis is complete and the environmental conditions are appropriate for spawning. PMID- 10456866 TI - Arrest of spermatogonial differentiation in jsd/jsd, Sl17H/Sl17H, and cryptorchid mice. AB - The nature of the spermatogenic arrest in cryptorchid C57Bl mice and in jsd/jsd and Sl17H/Sl17H mutant mice was identified by studying whole mounts of seminiferous tubules. In all three types of mice, virtually only A spermatogonia were found, topographically arranged in clones of 1 to 16 (rarely more) cells. These clonal sizes are typical for undifferentiated spermatogonia. The proportion of these cells lying in chains of more than 2 cells (50-70%) was comparable to that seen in epithelial stages VII-VIII in the normal epithelium. It is concluded that in all three types of mice, spermatogenesis is arrested at the point where the undifferentiated A spermatogonia, specifically A(al) spermatogonia, differentiate into the first generation of the differentiating-type spermatogonia, the A1 spermatogonia. The remaining A spermatogonia were proliferating, but no accumulation of spermatogonia was present, as spermatogonial apoptosis also took place. Spermatogonial clones of all sizes were seen to undergo apoptosis, but there were relatively many large apoptotic clones, indicating that the clones became more vulnerable when they became larger. In contrast to what is seen in the normal epithelium, odd-numbered clones, not composed of 2(n) cells, were present, as well as clumps of 2 or more spermatogonial nuclei in the same cytoplasm, in all three types of mice. This indicates a lack of integrity of spermatogonial clones, also observed in other situations with a relative paucity of cells on the basal membrane. It is concluded that the differentiation of the undifferentiated spermatogonia, affected in all three types of mice as well as in vitamin A-deficient animals, is a rather vulnerable point in the spermatogenic developmental pathway. PMID- 10456867 TI - Streptococcal pyrogenic exotoxin F (SpeF) causes permeabilization of lung blood vessels. AB - Acute respiration distress syndrome (ARDS) is a typical complication in toxic shock-like syndrome (TSLS) caused by Streptococcus pyogenes. An isolated perfused rat lung model was used to identify the causative agent of ARDS in TSLS in this study. Some crude preparations separated from the culture supernatants of S. pyogenes isolates caused rapid increases in the weight of perfused lungs, indicating vascular permeabilization. Six samples from M type 1 and 3 isolates from TSLS and pharyngitis patients showed strong permeabilization activity, whereas preparations from isolates of other M types (although the number of isolates examined was limited) were negative. The active substance was purified to a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis using various columns, and the N-terminal amino acid sequence was determined. The resultant sequence of eight amino acids was identical to SpeF (mitogenic factor). Moreover, the vascular permeabilization activity of the purified band was abolished with anti-SpeF antiserum prepared by immunizing with the purified SpeF. This activity was also neutralized by the antiserum prepared by immunizing with a synthetic peptide derived from the published SpeF sequence. These results suggested that streptococcal SpeF is a major cause of permeabilization of lung blood vessels and sufficient for the pathogenesis of ARDS under the conditions of TSLS caused by S. pyogenes. PMID- 10456868 TI - Identification of amino acid residues of the T-cell epitope of Mycobacterium tuberculosis alpha antigen critical for Vbeta11(+) Th1 cells. AB - Stimulation of Mycobacterium tuberculosis-primed lymph node cells from C57BL/6 mice with alpha antigen (also known as antigen 85B and MPT59) induced cell proliferation, production of interleukin 2 and gamma interferon, and expansion of Vbeta11(+) CD4(+) T cells in conjunction with antigen-presenting cells in an I A(b)-restricted manner. Using a series of 15-amino-acid peptides that overlapped each other by 5 amino acids and spanned the mature alpha antigen, we identified the antigenic epitope for alpha antigen-specific Vbeta11(+) Th1 cells. That peptide (peptide-25), which corresponds to amino acid residues 240 to 254 of alpha antigen, contains a motif that is conserved in I-A(b) and requires processing by antigen-presenting cells. Using peptide-25-reactive Vbeta11(+) T cell clones and substituted peptide-25 mutants, we determined which amino acid residues within peptide-25 were critical for T-cell receptor (TCR) recognition. Our results showed that the amino acid residues at positions 245, 246, 248, 250, and 251 are important for recognition of TCRVbeta11 and that residues at positions 244, 247, 249, and 252 are I-A(b) contact residues. We also observed that active immunization of C57BL/6 mice with peptide-25 can lead to decreased bacterial load in the lungs of M. tuberculosis H37Rv-infected mice. These results should provide us with a useful tool for delineating the regulation of Vbeta11(+) Th1-cell development during M. tuberculosis infection and for developing a vaccine inducing a Th1-dominant immune response. PMID- 10456869 TI - Intranasal immunization with heat-inactivated Streptococcus pneumoniae protects mice against systemic pneumococcal infection. AB - In order to study the mucosal and serum antibody response to polysaccharide encapsulated bacteria in mice, a preparation of heat-inactivated Streptococcus pneumoniae type 4 was administered, with and without cholera toxin, at various mucosal sites. It appeared that intranasal immunization of nonanesthesized animals was superior to either oral, gastric, or colonic-rectal antigen delivery with regard to the induction of serum immunoglobulin G (IgG) and IgA, as well as saliva IgA antibodies specific for pneumococci. The marked IgA antibody response in feces after intranasal, but not after oral or gastric, immunization is suggestive of a cellular link between the nasal induction site and the distant mucosal effector sites. Intranasal immunization also induced antibodies in serum and in mucosal secretions against type-specific capsular polysaccharide. IgA and IgG antibody levels in pulmonary lavage fluids correlated well with saliva IgA and serum IgG antibodies, respectively. Antibody determinations in pulmonary secretions may therefore be redundant in some cases, and the number of experimental animals may be reduced accordingly. After intraperitoneal challenge with type 4 pneumococci, mice immunized intranasally were protected against both systemic infection and death, even without the use of cholera toxin as a mucosal adjuvant. Thus, an efficient intranasal vaccine against invasive pneumococcal disease may be based on a very simple formulation with whole killed pneumococci. PMID- 10456870 TI - Fibrinogen cleavage by the Streptococcus pyogenes extracellular cysteine protease and generation of antibodies that inhibit enzyme proteolytic activity. AB - The extracellular cysteine protease from Streptococcus pyogenes is a virulence factor that plays a significant role in host-pathogen interaction. Streptococcal protease is expressed as an inactive 40-kDa precursor that is autocatalytically converted into a 28-kDa mature (active) enzyme. Replacement of the single cysteine residue involved in formation of the enzyme active site with serine (C192S mutation) abolished detectable proteolytic activity and eliminated autocatalytic processing of zymogen to the mature form. In the present study, we investigated activity of the wild-type (wt) streptococcal protease toward human fibrinogen and bovine casein. The former is involved in blood coagulation, wound healing, and other aspects of hemostasis. Treatment with streptococcal protease resulted in degradation of the COOH-terminal region of fibrinogen alpha chain, indicating that fibrinogen may serve as an important substrate for this enzyme during the course of human infection. Polyclonal antibodies generated against recombinant 40- and 28-kDa (r40- and r28-kDa) forms of the C192S streptococcal protease mutant exhibited high enzyme-linked immunosorbent assay titers but demonstrated different inhibition activities toward proteolytic action of the wt enzyme. Activity of the wt protease was readily inhibited when the reaction was carried out in the presence of antibodies generated against r28-kDa C192S mutant. Antibodies produced against r40-kDa C192S mutant had no significant effect on proteolysis. These data suggest that the presence of the NH(2)-terminal prosegment prevents generation of functionally active antibodies and indicate that inhibition activity of antibodies most likely depends on their ability to bind the active-site region epitope(s) of the protein. PMID- 10456871 TI - Group A Streptococcus induces apoptosis in human epithelial cells. AB - Internalization of group A streptococcus (GAS) by epithelial cells may have a role in causing invasive diseases. The purpose of this study was to examine the fate of GAS-infected epithelial cells. GAS has the ability to invade A-549 and HEp-2 cells. Both A-549 and HEp-2 cells were killed by infection with GAS. Epithelial cell death mediated by GAS was at least in part through apoptosis, as shown by changes in cellular morphology, DNA fragmentation laddering, and propidium iodide staining for hypodiploid cells. A total of 20% of A-549 cells and 11 to 13% of HEp-2 cells underwent apoptosis after 20 h of GAS infection, whereas only 1 to 2% of these cells exhibited spontaneous apoptosis. We further examined whether streptococcal pyrogenic exotoxin B (SPE B), a cysteine protease produced by GAS, was involved in the apoptosis of epithelial cells. The speB isogenic mutants had less ability to induce cell death than wild-type strains. When A-549 cells were cocultured with the mutant and SPE B for 2 h, the percentage of apoptotic cells did not increase although the number of intracellular bacteria increased to the level of wild-type strains. In addition, apoptosis was blocked by cytochalasin D treatment, which interfered with cytoskeleton function. The caspase inhibitors Z-VAD.FMK, Ac-YVAD.CMK, and Ac DEVD.FMK inhibited GAS-induced apoptosis. These results demonstrate for the first time that GAS induces apoptosis of epithelial cells and internalization is required for apoptosis. The caspase pathway is involved in GAS-induced apoptosis, and the expression of SPE B in the cells enhances apoptosis. PMID- 10456873 TI - Detection of intrastrain antigenic variation of Bacteroides fragilis surface polysaccharides by monoclonal antibody labelling. AB - Bacteroides fragilis is a constituent of the normal resident microbiota of the human intestine and is the gram-negative obligately anaerobic bacterium most frequently isolated from clinical infection. Surface polysaccharides are implicated as potential virulence determinants. We present evidence of within strain immunochemical variation of surface polysaccharides in populations that are noncapsulate by light microscopy as determined by monoclonal antibody labelling. Expression of individual epitopes can be enriched from a population of an individual strain by use of immunomagnetic beads. Also, individual colonies in which either >94% or <7% of the bacteria carry an individual epitope retain this level of expression when subcultured into broth. In broth cultures where >94% of the bacteria carry a given epitope, there is no enrichment for other epitopes recognized by different polysaccharide-specific monoclonal antibodies. This intrastrain variation has important implications for the development of potential vaccines or immunodiagnostic tests. PMID- 10456872 TI - CpG motifs in Porphyromonas gingivalis DNA stimulate interleukin-6 expression in human gingival fibroblasts. AB - We suggest here that Porphyromonas gingivalis DNA may function as a virulence factor in periodontal disease through expression of inflammatory cytokine. The bacterial DNA markedly stimulated in a dose-dependent manner interleukin-6 (IL-6) production by human gingival fibroblasts. The stimulatory action was eliminated by treatment with DNase but not RNase. The stimulatory effect was not observed in the fibroblasts treated with eucaryotic DNAs. The bacterial DNA also stimulated in dose- and treatment time-dependent manners the expression of the IL-6 gene in the cells. In addition, the stimulatory effect was eliminated when the DNA was methylated with CpG motif methylase. Interestingly, a 30-base synthetic oligonucleotide containing the palindromic motif GACGTC could stimulate expression of the IL-6 gene and production of its protein in the cells. Furthermore, the synthetic oligonucleotide-induced expression of this cytokine gene was blocked by pyrrolidine dithiocarbamate and N-acetyl-L-cystine, potent inhibitors of transcriptional factor NF-kappaB. Gel mobility shift assay showed increased binding of NF-kappaB to its consensus sequence in the synthetic oligonucleotide-treated cells. Also, using specific antibody against p50 and p65, which compose NF-kappaB, we showed the consensus sequence-binding proteins to be NF-kappaB. These results are the first to demonstrate that the internal CpG motifs in P. gingivalis DNA stimulate IL-6 expression in human gingival fibroblasts via stimulation of NF-kappaB. PMID- 10456874 TI - Host responses to recombinant hemagglutinin B of Porphyromonas gingivalis in an experimental rat model. AB - Porphyromonas gingivalis, a gram-negative, black-pigmented anaerobe, is among the microorganisms implicated in the etiology of adult periodontal disease. This bacterium possesses a number of factors, including hemagglutinins, of potential importance in virulence. Several hemagglutinin genes have been identified, cloned, and expressed in Escherichia coli. The purpose of this study was to characterize host responses to purified recombinant hemagglutinin B (rHag B), using the conventional Fischer rat as the experimental animal model. The effectiveness of immunization with rHag B on protection against experimental periodontal bone loss following infection with P. gingivalis was also evaluated. Groups of rats were immunized by the subcutaneous route with rHag B in complete Freund's adjuvant, immunized with rHag B and orally infected with P. gingivalis, nonimmunized and noninfected, or orally infected with P. gingivalis only. Serum and saliva samples were collected throughout the experiment and evaluated for serum immunoglobulin G (IgG) and IgM and salivary IgA antibody activity by enzyme linked immunosorbent assay. No salivary IgA anti-Hag B activity was detected in the various groups of rats. A slight serum IgM response similar to that seen in preimmune samples was observed. Serum IgG antibody activity to Hag B was detected only in samples from rats immunized with rHag B. This response was primarily of the IgG1 and IgG2a subclasses, followed by IgG2b and low levels of IgG2c. Supernatants from rHag B-stimulated splenic lymphoid cell cultures from immunized rats contained high levels of gamma interferon, followed by interleukin-2 (IL-2), IL-10, and then IL-4. These results are consistent with the induction of T helper type 1 (Th1)- and Th2-like responses. Western blot analysis of sera derived from rHag B-immunized rats reacted with trichloroacetic acid (TCA) precipitates of P. gingivalis 33277, 381, A7A1-28, and W50, revealing a 50-kDa band reflective of Hag B. However, sera derived from rats immunized with P. gingivalis whole cells or from rats infected with P. gingivalis only did not react with rHag B but did react with TCA precipitates of P. gingivalis strains. Finally, radiographic measurements of periodontal bone loss indicated that rats immunized with rHag B had less bone loss than those infected with P. gingivalis only. These results demonstrate the effectiveness of purified rHag B in inducing a protective immune response and support the potential usefulness of this component of P. gingivalis in the development of a vaccine against adult periodontitis. PMID- 10456875 TI - Intranasal administration of synthetic recombinant peptide-based vaccine protects mice from infection by Schistosoma mansoni. AB - Schistosomiasis is the cause of a chronic debilitating disease which accounts for significant mortality and morbidity every year, especially in tropical and subtropical areas. An epitope derived from the protective surface protein 9B-Ag of Schistosoma mansoni, designated 9B peptide-1, was previously showed to be protective in mice when conjugated to bovine serum albumin and administered subcutaneously in complete Freund's adjuvant. In this work, this protective peptide was expressed in the flagellin of a Salmonella vaccine strain, and the isolated recombinant flagella were used for immunization of mice. Since during the invasion of the parasite into the host the schistosomula migrate first to the lungs, the intranasal route of administration was employed in order to halt the parasite at an early stage of the infection. Such intranasal immunization with this peptide expressed in flagellin, without the addition of adjuvants, resulted in a significant humoral response and also led to protection against challenge infection, manifested as a reduction of the worm burden by an average of 42%. PMID- 10456876 TI - Characterization of the interaction between Yersinia enterocolitica biotype 1A and phagocytes and epithelial cells in vitro. AB - Yersinia enterocolitica strains of biotype 1A are increasingly being recognized as etiological agents of gastroenteritis. However, the mechanisms by which these bacteria cause disease differ from those of highly invasive, virulence plasmid bearing Y. enterocolitica strains and are poorly understood. We have investigated several biotype 1A strains of diverse origin for their ability to resist killing by professional phagocytes. All strains were rapidly killed by polymorphonuclear leukocytes but persisted within macrophages (activated with gamma interferon) to a significantly greater extent (survival = 40.5% +/- 17.4%) than did Escherichia coli HB101 (9.3% +/- 0.7%; P = 0.0001). Strains isolated from symptomatic patients were significantly more resistant to killing by macrophages (survival = 48.9% +/- 19.5%) than were strains obtained from food or the environment (survival = 32.1% +/- 10.3%; P = 0.04). Some strains which had been ingested by macrophages or HEp-2 epithelial cells showed a tendency to reemerge into the tissue culture medium over a period lasting several hours. This phenomenon, which we termed "escape," was observed in 14 of 15 strains of clinical origin but in only 3 of 12 nonclinical isolates (P = 0.001). The capacity of bacteria to escape from cells was not directly related to their invasive ability. To determine if escape was due to host cell lysis, we used a variety of techniques, including lactate dehydrogenase release, trypan blue exclusion, and examination of infected cells by light and electron microscopy, to measure cell viability and lysis. These studies established that biotype 1A Y. enterocolitica strains were able to escape from macrophages or epithelial cells without causing detectable cytolysis, suggesting that escape was achieved by a process resembling exocytosis. The observations that biotype 1A Y. enterocolitica strains of clinical origin are significantly more resistant to killing by macrophages and significantly more likely to escape from host cells than are strains of nonclinical origin suggest that these properties may account for the virulence of these bacteria. PMID- 10456877 TI - Truncated surface protective antigen (SpaA) of Erysipelothrix rhusiopathiae serotype 1a elicits protection against challenge with serotypes 1a and 2b in pigs. AB - Erysipelothrix rhusiopathiae is a causal agent of swine erysipelas, which is of economic importance in the swine industry by virtue of causing acute septicemia, chronic arthritis, and endocarditis. However, little is known about the genetic properties of its protective antigens. Recently, a surface protective antigen (SpaA) gene was identified from serotype 2 in a mouse model. We cloned spaA from virulent strain Fujisawa (serotype 1a) and determined that the N-terminal 342 amino acids without C-terminal repeats of 20 amino acids have the ability to elicit protection in mice. Fusions of 342 amino acids of Fujisawa SpaA and histidine hexamer (HisSpa1.0) protected pigs against challenge with both serotype 1 and serotype 2, the most important serotypes in the swine industry. Pigs immunized with HisSpa1.0 reacted well with both HisSpa1.0 and intact SpaA by enzyme-linked immunosorbent assay and immunoblotting. Serum collected at the time of challenge from a pig immunized with HisSpa1. 0 markedly enhanced the in vitro phagocytic and killing activity of pig neutrophils against the bacteria. DNA sequences of protective regions of spaA genes from five strains of serotypes 1 and 2 were almost identical. The full DNA sequences also seemed to be conserved among strains of all 12 serotype reference strains harboring the spaA gene by restriction fragment length polymorphism analysis of PCR products. These results indicates that SpaA is a common protective antigen of serotypes 1 and 2 of E. rhusiopathiae in swine and will be a useful tool for development of new types of vaccines and diagnostic tools for effective control of the disease. PMID- 10456878 TI - Protective role for interleukin-5 during chronic Toxoplasma gondii infection. AB - To investigate the role of interleukin-5 (IL-5) during Toxoplasma gondii infection, IL-5 knockout (KO) mice and C57BL/6 control mice were infected intraperitoneally with ME49 cysts and the course of infection was monitored. The mortality rate during chronic infection was significantly greater in IL-5 deficient animals, and consistent with this finding, the KO mice harbored a greater number of brain cysts and tachyzoites than did their wild-type counterparts. Although the IL-5 KO animals did not succumb until late during infection, increased susceptibility, as measured by accelerated weight loss, was detectable during the acute stages of infection. The amounts of total immunoglobulin (Ig), IgM, and IgG2b were comparable in both strains, while the amount of IgG1 was much smaller in IL-5 KO mice. Spleen cell production of IL-12 in response to T. gondii antigen was approximately threefold lower in the KO strain, and this decrease correlated with a selective loss of B lymphocytes during culture. A link between the presence of B cells and augmented IL-12 production was established by the finding that after removal of B cells with monoclonal antibody and complement, wild-type- and KO-derived cells produced equivalent levels of IL-12 in response to T. gondii antigen. These results demonstrate a protective role of IL-5 against T. gondii infection and suggest that IL-5 may play a role in the production of IL-12. PMID- 10456880 TI - Genetically detoxified mutants of heat-labile toxin from Escherichia coli are able to act as oral adjuvants. AB - Detoxified mutants of the Escherichia coli heat-labile toxin (LT) act as mucosal adjuvants to intranasally presented coadministered antigens. Here, we compare the adjuvant activity of a panel of detoxified derivatives of LT, using both intranasal (i.n.) and oral (p.o.) routes of administration. The mutants used as adjuvants varied in sensitivity to proteases and toxicity. With keyhole limpet hemocyanin (KLH) as the bystander antigen, the immune responses to i. n. immunizations were consistently higher than the equivalent p.o. -delivered proteins. LT-G192, a mutant which demonstrates a 10-fold reduction in toxicity in vitro, demonstrated wild-type adjuvant activity both i.n. and p.o., inducing similar titers of KLH specific antibody in the sera and immunoglobulin A in local mucosal secretions as wild-type LT. In line with previous data, the nontoxic holotoxoid LT-K63 induced intermediate immune responses in both the serum and mucosal secretions which were lower than those achieved with wild-type LT but at least 10-fold higher than those measured when the antigen was administered with LT-B. Although significant levels of local and systemic anti-KLH antibodies were induced following p.o. immunization with LT-K63, cellular proliferative responses to KLH was poor or undetectable. In contrast, LT and LT-G192 induced significant T-cell responses to KLH following p.o. immunization. These proliferating cells secreted both gamma interferon and interleukin-5, suggesting that the type of immune response induced following p.o. coimmunization with LT and purified protein is a mixed Th1/Th2 response. PMID- 10456879 TI - Characterization of binding of adenylate cyclase toxin to target cells by flow cytometry. AB - Adenylate cyclase (AC) toxin from Bordetella pertussis intoxicates eukaryotic cells by increasing intracellular cyclic AMP (cAMP) levels. In addition, insertion of AC toxin into the plasma membrane causes efflux of intracellular K(+) and, in a related process, hemolysis of sheep erythrocytes. Although intoxication, K(+) efflux, and hemolysis have been thoroughly investigated, there is little information on the nature of the interaction of this toxin with intact target cells. Using flow cytometry, we observe that binding of AC toxin to sheep erythrocytes and Jurkat T lymphocytes is dependent on posttranslational acylation of the toxin. Extracellular calcium is also necessary, with a steep calcium concentration dependence similar to that required for intoxication and hemolysis. Binding of AC toxin is concentration dependent but unsaturable up to 50 micrograms/ml, suggesting that if there is a specific receptor molecule with which the toxin interacts, it is not limiting. Visualization of cells by fluorescence microscopy supports the data obtained by flow cytometry and reveals a peripheral pattern of toxin distribution. AC toxin binds to erythrocytes at both 0 and 37 degrees C; however, the total binding at 0 degrees C is less than that at 37 degrees C. In human erythrocytes, AC toxin does not cause an increase in K(+) efflux or hemolysis. While AC toxin exhibits reduced potency to increase cAMP in these cells than in sheep erythrocytes, there is only a modest reduction in the binding of the toxin as measured by flow cytometry. Further use of this technique will provide new approaches for dynamic and functional analysis of the early steps involved in intoxication, K(+) efflux, and hemolysis produced by AC toxin. PMID- 10456881 TI - Cloning and molecular characterization of plasmid-encoded antigens of Borrelia burgdorferi. AB - Thirteen independent clones that encode Borrelia burgdorferi antigens utilizing antiserum from infection-immune rabbits were identified. The serum was adsorbed against noninfectious B. burgdorferi B31 to enrich for antibodies directed against either infection-associated antigens of B. burgdorferi B31 or proteins preferentially expressed during mammalian infection. The adsorption efficiency of the immune rabbit serum (IRS) was assessed by Western immunoblot analysis with protein lysates derived from infectious and noninfectious B. burgdorferi B31. The adsorbed IRS was used to screen a B. burgdorferi expression library to identify immunoreactive phage clones. Clones were then expressed in Escherichia coli and subsequently analyzed by Western blotting to determine the molecular mass of the recombinant B. burgdorferi antigens. Southern blot analysis of the 13 clones indicated that 10 contained sequences unique to infectious B. burgdorferi. Nucleotide sequence analysis indicated that the 13 clones were composed of 9 distinct genetic loci and that all of the genes identified were plasmid encoded. Five of the clones carried B. burgdorferi genes previously identified, including those encoding decorin binding proteins A and B (dbpAB), a rev homologue present on the 9-kb circular plasmid (cp9), a rev homologue from the 32-kb circular plasmid (cp32-6), erpM, and erpX. Additionally, four previously uncharacterized loci with no known homologues were identified. One of these unique clones encoded a 451-amino-acid lipoprotein with 21 consecutive, invariant 9-amino-acid repeats near the amino terminus that we have designated VraA (for "virulent strain associated repetitive antigen A"). Since all the antigens identified are recognized by serum from infection immune rabbits, these antigens represent potential vaccine candidates and, based on the identification of dbpAB in this screen, may also be involved in pathogenic processes operative in Lyme borreliosis. PMID- 10456882 TI - Down-regulation of Th2 responses by Brucella abortus, a strong Th1 stimulus, correlates with alterations in the B7.2-CD28 pathway. AB - Down-regulation of the Th2-like response induced by ovalbumin-alum (OVA/alum) immunization by heat-killed Brucella abortus was not reversed by anti-IL-12 antibody treatment or in gamma interferon (IFN-gamma) knockout mice, suggesting that induction of Th1 cytokines was not the only mechanism involved in the B. abortus-mediated inhibition of the Th2 response to OVA/alum. The focus of this study was to determine whether an alternative pathway involves alteration in expression of costimulatory molecules. First we show that the Th2-like response to OVA/alum is dependent on B7.2 interaction with ligand since it can be abrogated by anti-B7.2 treatment. Expression of costimulatory molecules was then studied in mice immunized with OVA/alum in the absence or presence of B. abortus. B7.2, but not B7.1, was up-regulated on mouse non-T and T cells following immunization with B. abortus. Surprisingly, B. abortus induced down-regulation of CD28 and up-regulation of B7.2 on murine CD4(+) and CD8(+) T cells. These effects on T cells were maximal for CD28 and B7.2 at 40 to 48 h and were not dependent on interleukin-12 (IL-12) or IFN-gamma. On the basis of these results, we propose that the IL-12/IFN-gamma-independent inhibition of Th2 responses to OVA/alum is secondary to the effects of B. abortus on expression of costimulatory molecules on T cells. We suggest that down-regulation of CD28 following activation inhibits subsequent differentiation of Th0 into Th2 cells. In addition, decreased expression of CD28 and increased expression of B7.2 on T cells would favor B7.2 interaction with CTLA-4 on T cells, and this could provide a negative signal to developing Th2 cells. PMID- 10456883 TI - Intracellular growth in Acanthamoeba castellanii affects monocyte entry mechanisms and enhances virulence of Legionella pneumophila. AB - Since Legionella pneumophila is an intracellular pathogen, entry into and replication within host cells are thought to be critical to its ability to cause disease. L. pneumophila grown in one of its environmental hosts, Acanthamoeba castellanii, is phenotypically different from L. pneumophila grown on standard laboratory medium (BCYE agar). Although amoeba-grown L. pneumophila displays enhanced entry into monocytes compared to BCYE-grown bacteria, the mechanisms of entry used and the effects on virulence have not been examined. To explore whether amoeba-grown L. pneumophila differs from BCYE-grown L. pneumophila in these characteristics, we examined entry into monocytes, replication in activated macrophages, and virulence in mice. Entry of amoeba-grown L. pneumophila into monocytes occurred more frequently by coiling phagocytosis, was less affected by complement opsonization, and was less sensitive to microtubule and microfilament inhibitors than was entry of BCYE-grown bacteria. In addition, amoeba-grown L. pneumophila displays increased replication in monocytes and is more virulent in A/J, C57BL/6 Beige, and C57BL/6 mice. These data demonstrate for the first time that the intra-amoebal growth environment affects the entry mechanisms and virulence of L. pneumophila. PMID- 10456884 TI - A defect in interleukin-10 leads to enhanced malarial disease in Plasmodium chabaudi chabaudi infection in mice. AB - Infection of interleukin-10 (IL-10)-nonexpressing (IL-10(-/-)) mice with Plasmodium chabaudi chabaudi (AS) leads to exacerbated pathology in female mice and death in a proportion of them. Hypoglycemia, hypothermia, and loss in body weight were significantly greater in female IL-10(-/-) mice than in male knockout mice and all wild-type (WT) mice during the acute phase of infection. At this time, both female and male IL-10(-/-) mice produced more gamma interferon (IFN gamma), tumor necrosis factor alpha (TNF-alpha), and IL-12p40 mRNA than their respective WT counterparts. Inactivation of IFN-gamma in IL-10(-/-) mice by the injection of anti-IFN-gamma antibodies or by the generation of IL-10(-/-) IFN gamma receptor(-/-) double-knockout mice resulted in reduced mortality but did not affect body weight, temperature, or blood glucose levels. The data suggest that IFN-gamma-independent pathways may be responsible for these pathological features of P. chabaudi malaria and may be due to direct stimulation of TNF-alpha by the parasite. Since male and female knockout mice both produce more inflammatory cytokines than their WT counterparts, it is likely that the mortality seen in females is due to the nature or magnitude of the response to these cytokines rather than the amount of IFN-gamma or TNF-alpha produced. PMID- 10456885 TI - Role of ornibactin biosynthesis in the virulence of Burkholderia cepacia: characterization of pvdA, the gene encoding L-ornithine N(5)-oxygenase. AB - Burkholderia cepacia is a frequent cause of respiratory infections in cystic fibrosis patients. B. cepacia has been shown to produce at least four siderophores which may play a role in the virulence of this organism. To characterize genes involved in the synthesis of siderophores, Tn5-OT182 mutants were isolated in strain K56-2, which produces two siderophores, salicylic acid (SA) and ornibactins. Two mutants were characterized that did not produce zones on Chrome Azurol S agar in a commonly used assay to detect siderophore activity. These mutants were determined to produce sevenfold more SA than K56-2 yet did not produce detectable amounts of ornibactins. These mutants, designated I117 and T10, had a transposon insertion in genes with significant homology to pyoverdine biosynthesis genes of Pseudomonas aeruginosa. I117 contained an insertion in a pvdA homolog, the gene for the enzyme L-ornithine N(5)-oxygenase, which catalyzes the hydroxylation of L-ornithine. Ornibactin synthesis in this mutant was partially restored when the precursor L-N(5)-OH-Orn was added to the culture medium. T10 contained an insertion in a pvdD homolog, which is a peptide synthetase involved in pyoverdine synthesis. beta-Galactosidase activity was iron regulated in both I117 and T10, suggesting that the transposon was inserted downstream of an iron-regulated promoter. Tn5-OT182 contains a lacZ gene that is expressed when inserted downstream of an active promoter. Both I117 and T10 were deficient in uptake of iron complexed to either ornibactins or SA, suggesting that transposon insertions in ornibactin biosynthesis genes also affected other components of the iron transport mechanism. The B. cepacia pvdA homolog was approximately 47% identical and 59% similar to L-ornithine N(5)-oxygenase from P. aeruginosa. Three clones were identified from a K56-2 cosmid library that partially restored ornibactin production, SA production, and SA uptake to parental levels but did not affect the rate of (59)Fe-ornibactin uptake in I117. A chromosomal pvdA deletion mutant was constructed that had a phenotype similar to that of I117 except that it did not hyperproduce SA. The pvdA mutants were less virulent than the parent strain in chronic and acute models of respiratory infection. A functional pvdA gene appears to be required for effective colonization and persistence in B. cepacia lung infections. PMID- 10456886 TI - Role of Mycoplasma penetrans endonuclease P40 as a potential pathogenic determinant. AB - Recently, we reported the purification to homogeneity and characterization of Ca(2+)- and Mg(2+)-dependent endonuclease P40 produced by Mycoplasma penetrans (M. Bendjennat, A. Blanchard, M. Loutfi, L. Montagnier, and E. Bahraoui, J. Bacteriol. 179; 2210-2220, 1997), a mycoplasma which was isolated for the first time from the urine of human immunodeficiency virus-infected patients. To evaluate how this nuclease could interact with host cells, we tested its effect on CEM and Molt-4 lymphocytic cell lines and on peripheral blood mononuclear cells. We observed that 10(-7) to 10(-9) M P40 is able to mediate a cytotoxic effect. We found that 100% of cells were killed after 24 h of incubation with 10( 7) M P40 while only 40% cytotoxicity was obtained after 72 h of incubation with 10(-9) M P40. Phase-contrast microscopy observations of P40-treated cells revealed morphological changes, including pronounced blebbing of the plasma membrane and cytoplasmic shrinkage characteristic of programmed cell death, which is in agreement with the internucleosomal fragmentation of P40-treated cell DNA as shown by agarose gel electrophoresis. We showed that (125)I-radiolabeled or fluorescein isothiocyanate-labeled P40 was able to bind specifically in a dose dependent manner to the cell membrane of CEM cells, which suggested that the cytotoxicity of P40 endonuclease was mediated by its interaction with the cell surface receptor(s). The concentration of unlabeled P40 required to inhibit by 50% the formation of (125)I-P40-CEM complexes was about 3 x 10(-9) M, indicating a high-affinity interaction. Both P40 interaction and cytotoxicity are Ca(2+) dependent. Our results suggest that the cytotoxicity of M. penetrans observed in vitro is mediated at least partially by secreted P40, which, after interaction with host cells, can induce an apoptosis-like death. These results strongly suggest a major role of mycoplasmal nucleases as potential pathogenic determinants. PMID- 10456887 TI - Sulfated polysaccharide-directed recruitment of mammalian host proteins: a novel strategy in microbial pathogenesis. AB - Fundamental to the virulence of microbial pathogens is their capacity for adaptation and survival within variable, and often hostile, environments encountered in the host. We describe a novel, extragenomic mechanism of surface modulation which may amplify the adaptive and pathogenic potential of numerous bacterial species, including Staphylococcus, Yersinia, and pathogenic Neisseria species, as well as Helicobacter pylori and Streptococcus pyogenes. The mechanism involves specific bacterial recruitment of heparin, glycosaminoglycans, or related sulfated polysaccharides, which in turn serve as universal binding sites for a diverse array of mammalian heparin binding proteins, including adhesive glycoproteins (vitronectin and fibronectin), inflammatory (MCP-3, PF-4, and MIP 1alpha) and immunomodulatory (gamma interferon) intermediates, and fibroblast growth factor. This strategy impacts key aspects of microbial pathogenicity as exemplified by increased bacterial invasion of epithelial cells and inhibition of chemokine-induced chemotaxis. Our findings illustrate a previously unrecognized form of parasitism that complements classical virulence strategies encoded within the microbial genome. PMID- 10456888 TI - Molecular and idiotypic analyses of the antibody response to Cryptococcus neoformans glucuronoxylomannan-protein conjugate vaccine in autoimmune and nonautoimmune mice. AB - The antibody response to Cryptococcus neoformans capsular glucuronoxylomannan (GXM) in BALB/c mice frequently expresses the 2H1 idiotype (Id) and is restricted in variable gene usage. This study examined the immunogenicity of GXM-protein conjugates, V (variable)-region usage, and 2H1 Id expression in seven mouse strains: BALB/c, C57BL/6, A/J, C3H, NZB, NZW, and (NZB x NZW)F(1) (NZB/W). All mouse strains responded to vaccination with GXM conjugated to tetanus toxoid (TT), the relative magnitude of the antibody response being BALB/c approximately C3H > C57BL/6 approximately NZB approximately NZW approximately NZB/W > A/J. Analysis of serum antibody responses to GXM with polyclonal and monoclonal antibodies to the 2H1 Id revealed significant inter- and intrastrain differences in idiotype expression. Thirteen monoclonal antibodies (MAbs) (two immunoglobulin M [IgM], three IgG3, one IgG1, three IgG2a, two IgG2b, and two IgA) to GXM were generated from one NZB/W mouse and one C3H/He mouse. The MAbs from the NZB/W mouse were all 2H1 Id positive (Id(+)) and structurally similar to those previously generated in BALB/c mice, including the usage of a V(H) from the 7183 family and Vkappa5.1. Administration of both 2H1 Id(+) and Id(-) MAbs from NZB/W and C3H/H3 mice prolonged survival in a mouse model of cryptococcosis. Our results demonstrate (i) that V-region restriction as indicated by the 2H1 Id is a feature of both primary and secondary responses of several mouse strains; and (ii) that there is conservation of V-region usage and length of the third complementarity-determining region in antibodies from three mouse strains. The results suggest that V-region restriction is a result of antibody structural requirements necessary for binding an immunodominant antigen in GXM. PMID- 10456889 TI - Interaction of Leishmania gp63 with cellular receptors for fibronectin. AB - The most abundant protein on the surface of the promastigote form of the protozoan parasites Leishmania spp. is a 63-kDa molecule, designated gp63 or leishmanolysin. Because gp63 has been shown to possess fibronectin-like properties, we examined the interaction of gp63 with the cellular receptors for fibronectin. We measured the direct binding of Leishmania to human macrophages or to transfected mammalian cells expressing human fibronectin receptors. Leishmania expressing gp63 exhibited modest but reproducible adhesion to human macrophages and to transfected CHO cells expressing alpha4/beta1 fibronectin receptors. In both cases, this interaction depended on gp63 but occurred independently of the SRYD sequence of gp63, because parasites expressing gp63 with a mutated SRYD sequence bound to macrophages and alpha4/beta1 receptor-expressing cells as well as did wild-type parasites. The contribution of gp63 to parasite adhesion was more pronounced when the assays were performed in the presence of complement, suggesting that the receptors for complement and fibronectin may cooperate to mediate the efficient adhesion of parasites to macrophages. The interaction of gp63 with fibronectin receptors may also play an important role in parasite internalization by macrophages. Erythrocytes to which gp63 was cross-linked were efficiently phagocytized by macrophages, whereas control erythrocytes opsonized with complement alone bound to macrophages but remained peripherally attached to the outside of the cell. Similarly, parasites expressing wild-type gp63 were rapidly and efficiently phagocytized by resting macrophages, whereas parasites lacking gp63 were internalized more slowly. This rapid internalization of gp63 expressing parasites was dependent on the beta1 integrins, because pretreatment of macrophages with monoclonal antibodies to the beta1 integrins decreased the internalization of gp63-expressing parasites. These observations indicate that complement receptors are the primary mediators of parasite adhesion; however, maximal parasite adhesion and internalization may require the participation of the beta1 integrins, which recognize fibronectin-like molecules such as gp63 on the surface of the parasite. PMID- 10456890 TI - Secretory leukocyte protease inhibitor interferes with uptake of lipopolysaccharide by macrophages. AB - Macrophages are among the most sensitive targets of bacterial endotoxin (LPS), responding to minute amounts of LPS by releasing a battery of inflammatory mediators. Transfection of macrophages with secretory leukocyte protease inhibitor (SLPI) renders these cells refractory to LPS stimulation. Here we show that uptake of LPS from soluble CD14 (sCD14)-LPS complexes by SLPI-overexpressing cells was only 50% of that seen in control cells. SLPI transfectants and mock transfectants did not differ in the surface expression of CD14 or CD18. We show, in addition, that recombinant human SLPI can bind to purified endotoxin in vitro. SLPI caused a decrease in the binding of LPS to sCD14 as assessed both by fluorescence quenching of labeled LPS and by nondenaturing polyacrylamide gel electrophoresis. These results suggest that the inhibitory effect of SLPI on macrophage responses to LPS may, in part, be due to its blockade of LPS transfer to soluble CD14 and its interference with uptake of LPS from LPS-sCD14 complexes by macrophages. PMID- 10456891 TI - Legionella pneumophila invasion of MRC-5 cells induces tyrosine protein phosphorylation. AB - After uptake and intracellular multiplication of Legionella pneumophila in MRC-5 lung fibroblasts, important cytoskeletal filament structures, like actin, tubulin, or vimentin, and a cell membrane-associated fibronectin were rearranged during early infection, resulting in a loss of cell adhesion and collapse of the cytoskeleton. Dysregulation of the cellular phosphorylation and dephosphorylation cascade may contribute to the observed changes and may support intracellular survival and multiplication of L. pneumophila. We therefore studied expression of phosphoproteins during intracellular growth of L. pneumophila. By using an anti tyrosine phosphoprotein antibody we showed that proteins phosphorylated on tyrosine residues accumulated progressively during late infection exclusively around or in phagosomes filled with bacteria. In contrast, expression of serine/threonine phosphoproteins did not change. To discern the origin of phosphorylated proteins, the host cells were treated with cycloheximide, an inhibitor of eukaryotic protein synthesis. The newly synthesized proteins were labeled metabolically with [(35)S]methionine-cysteine and immunoprecipitated with a phosphotyrosine-specific antibody. Sodium dodecyl sulfate gel electrophoresis gave evidence for synthesis of at least three protein clusters (160 to 200, 35 to 60, and 19 to 28 kDa) of Legionella origin that were phosphorylated on tyrosine residues 24 h after infection. Treatment of infected host cells with genistein, a tyrosine kinase inhibitor, revealed that tyrosine protein phosphorylation was not important for bacterial uptake but contributed to intracellular growth of L. pneumophila. Bacterial tyrosine phosphoproteins and the observed intracellular structural changes may be important to understanding the process involved in intracellular growth of L. pneumophila. PMID- 10456893 TI - Stress-induced membrane association of the Streptococcus mutans GTP-binding protein, an essential G protein, and investigation of its physiological role by utilizing an antisense RNA strategy. AB - SGP (for Streptococcus GTP-binding protein) is a Streptococcus mutans essential GTPase which has significant sequence identity to the previously identified Escherichia coli Era protein and to numerous other prokaryotic GTPase proteins of unknown function. Recent studies in our laboratory have addressed the possible role of SGP in the stress response of the oral pathogen S. mutans. Here we report that during growth in the early stationary phase, and in response to elevated temperatures or acidic pH, the distribution of SGP between the cytoplasm and the membranes of S. mutans cells varies. Immunoblot analysis of soluble and membrane protein fractions collected from the mid-log and early stationary growth phases of bacterial populations grown at normal temperature (37 degrees C) and at the elevated temperature of 43 degrees C, or at acidic pH, demonstrated that the total amount of SGP increased with the age of the bacterial culture, elevated temperature, or acidic pH. Furthermore, it was established that a substantial amount of SGP is associated with the membrane fraction under stress conditions. In order to investigate the physiological role of SGP, we constructed an S. mutans strain capable of chromosomal sgp antisense RNA expression, which interferes with the normal information processing of the sgp gene. Utilizing this strain, we determined conditions whereby the streptococcal cells can be depleted of SGP, thus avoiding the problem of constructing a conditional lethal system. From the results of measurements of the nucleotide pools extracted from the antisense strain and its isogenic counterpart, we propose that one of the physiological roles of SGP is regulation and modulation of the GTP/GDP ratio under different growth conditions. Moreover, we observed that in SGP-depleted cells the levels of glucan-binding protein A (GbpA) substantially increased, suggesting that GbpA may have stress response-related physiological functions. Finally, the potential applications of the antisense RNA approach that we employed are discussed. PMID- 10456892 TI - Invasive ability of an Escherichia coli strain isolated from the ileal mucosa of a patient with Crohn's disease. AB - Crohn's disease (CD) is an inflammatory bowel disease in which Escherichia coli strains have been suspected of being involved. We demonstrated previously that ileal lesions of CD are colonized by E. coli strains able to adhere to intestinal Caco-2 cells but devoid of the virulence genes so far described in the pathogenic E. coli strains involved in gastrointestinal infections. In the present study we compared the invasive ability of one of these strains isolated from an ileal biopsy of a patient with CD, strain LF82, with that of reference enteroinvasive (EIEC), enteropathogenic (EPEC), enterotoxigenic (ETEC), enteraggregative (EAggEC), enterohemorrhagic (EHEC), and diffusely adhering (DAEC) E. coli strains. Gentamicin protection assays showed that E. coli LF82 was able to efficiently invade HEp-2 cells. Its invasive level was not significantly different from that of EIEC and EPEC strains (P > 0.5) but significantly higher than that of ETEC (P < 0.03), EHEC (P < 0. 005), EAggEC (P < 0.004) and DAEC (P < 0.02) strains. Strain LF82 also demonstrated efficient ability to invade intestinal epithelial cultured Caco-2, Intestine-407, and HCT-8 cells. Electron microscopy examination of infected HEp-2 cells revealed the presence of numerous intracellular bacteria located in vacuoles or free in the host cell cytoplasm. In addition, the interaction of strain LF82 with epithelial cells was associated with the elongation of microvillar extensions that extruded from the host cell membranes and engulfed the bacteria. This internalization mechanism strongly resembles Salmonella- or Shigella-induced macropinocytosis. The use of cytochalasin D and colchicine showed that the uptake of strain LF82 by HEp-2 cells was mediated by both an actin microfilament-dependent mechanism and microtubule involvement. In addition, strain LF82 survived for at least 24 h in HEp-2 and Intestine-407 cells and efficiently replicated intracellularly in HEp-2 cells. PCR and hybridization experiments did not reveal the presence of any of the genetic determinants encoding EIEC, EPEC, or ETEC proteins involved in bacterial invasion. Thus, these findings show that LF82, which colonized the ileal mucosa of a patient with CD, is a true invasive E. coli strain and suggest the existence of a new potentially pathogenic group of E. coli, which we propose be designated adherent-invasive E. coli. PMID- 10456894 TI - Resistance to both complement activation and phagocytosis in type 3 pneumococci is mediated by the binding of complement regulatory protein factor H. AB - To study the role of surface-associated proteins in the virulence of Streptococcus pneumoniae, we used two serotype 3 strains, ATCC 6303 and WU2, and two PspA-negative mutants of WU2, an encapsulated one, JY1123 (Caps(+)/PspA(-)), and an unencapsulated one, DW3.8 (Caps(-)/PspA(-)). ATCC 6303 and WU2 were highly virulent in mice, while the virulence of JY1123 was slightly decreased (50% lethal doses [LD(50)s], 24, 6, and 147 CFU/mouse, respectively); DW3.8 was avirulent (LD(50), 2 x 10(8) CFU). In vitro, ATCC 6303, WU2, and JY1123 (Caps(+)/PspA(-)) strongly resisted complement activation and complement dependent opsonophagocytosis, whereas DW3.8 (Caps(-)/PspA(-)) was easily phagocytized in fresh serum. Trypsin treatment of ATCC 6303, WU2, and JY1123 (Caps(+)/PspA(-)) resulted in enhanced complement activation and complement dependent opsonophagocytosis. Trypsin had no deleterious effect on the polysaccharide capsule. In addition, trypsin pretreatment of ATCC 6303 strongly reduced virulence upon intraperitoneal challenge in mice. This indicated that surface proteins play a role in the resistance to complement activation and opsonophagocytosis and contribute to the virulence of type 3 pneumococci. In subsequent experiments, we could show that the modulation of complement activation was associated with surface components that bind complement regulator factor H; binding is trypsin sensitive and independent of prior complement activation. Immunoblotting of cell wall proteins of the virulent strain ATCC 6303 with anti-human factor H antibody revealed three factor H-binding proteins of 88, 150, and 196 kDa. Immunogold electron microscopy showed a close association of factor H-binding components with the outer surface of the cell wall. The role of these factor H-binding surface proteins in the virulence of pneumococci is interesting and warrants further investigation. PMID- 10456895 TI - Functional studies of a fibrinogen binding protein from Staphylococcus epidermidis. AB - A gene encoding a fibrinogen binding protein from Staphylococcus epidermidis was previously cloned, and the nucleotide sequence was determined. A portion of the gene encompassing the fibrinogen binding domain has now been subcloned in an expression-fusion vector. The fusion protein can bind to fibrinogen in a capture enzyme-linked immunosorbent assay and can be purified by fibrinogen affinity chromatography. This protein can completely inhibit the adherence of S. epidermidis to immobilized fibrinogen, suggesting that the adherence of S. epidermidis to fibrinogen is mainly due to this protein. Antibodies against this fibrinogen binding protein were also found to efficiently block the adherence of S. epidermidis to immobilized fibrinogen. Despite homology with clumping factors A and B from S. aureus (cell surface-associated proteins binding to fibrinogen), binding involved the beta chain of fibrinogen rather than the gamma chain, as in clumping factor A. PMID- 10456896 TI - Reactivation of latent tuberculosis: variations on the Cornell murine model. AB - Mycobacterium tuberculosis causes active tuberculosis in only a small percentage of infected persons. In most cases, the infection is clinically latent, although immunosuppression can cause reactivation of a latent M. tuberculosis infection. Surprisingly little is known about the biology of the bacterium or the host during latency, and experimental studies on latent tuberculosis suffer from a lack of appropriate animal models. The Cornell model is a historical murine model of latent tuberculosis, in which mice infected with M. tuberculosis are treated with antibiotics (isoniazid and pyrazinamide), resulting in no detectable bacilli by organ culture. Reactivation of infection during this culture-negative state occurred spontaneously and following immunosuppression. In the present study, three variants of the Cornell model were evaluated for their utility in studies of latent and reactivated tuberculosis. The antibiotic regimen, inoculating dose, and antibiotic-free rest period prior to immunosuppression were varied. A variety of immunosuppressive agents, based on immunologic factors known to be important to control of acute infection, were used in attempts to reactivate the infection. Although reactivation of latent infection was observed in all three variants, these models were associated with characteristics that limit their experimental utility, including spontaneous reactivation, difficulties in inducing reactivation, and the generation of altered bacilli. The results from these studies demonstrate that the outcome of Cornell model-based studies depends critically upon the parameters used to establish the model. PMID- 10456897 TI - Replication-defective adenovirus infection reduces Helicobacter felis colonization in the mouse in a gamma interferon- and interleukin-12-dependent manner. AB - Helicobacter infection leads to chronic inflammation of the stomach. Although the infection persists in spite of an immune response, animal studies have shown that adjuvant-based oral vaccines can protect against infection and even eliminate established infection. These vaccines are thought to induce a Th2 immune response, counterbalancing the Th1 response seen with natural infections. As a prelude to using adenovirus vectors carrying cytokine genes to modulate the immune response to established Helicobacter felis infection, we first examined the effect of the replication-defective adenovirus (RDA) vector itself. C57BL/6 mice chronically infected with H. felis (8 to 10 weeks) received intramuscular injections of RDA. The effect of RDA on the severity of H. felis colonization and the degree of gastric inflammation was assessed 2 weeks later. RDA caused a significant decrease in H. felis colonization without significantly altering the associated inflammation. RDA did not alter the H. felis-specific immunoglobulin G1 (IgG1), IgG2a, and IgA responses in the serum but was associated with an increase in gamma interferon (IFN-gamma)-producing CD8(+) spleen cells. To determine if IFN-gamma or Th1 cytokines were involved in the response to RDA, we examined RDA treatment of H. felis infection in mice lacking either IFN-gamma or interleukin-12 (IL-12). RDA failed to alter H. felis colonization in either of these two mouse strains. Thus, viral infection of mice chronically infected with H. felis led to a significant decrease in H. felis colonization in an IFN-gamma- and IL-12-dependent manner. These results demonstrate that Th1 responses associated with systemic viral infection can influence an established H. felis infection. PMID- 10456899 TI - The autolysin-encoding gene (lytA) of Streptococcus pneumoniae displays restricted allelic variation despite localized recombination events with genes of pneumococcal bacteriophage encoding cell wall lytic enzymes. AB - The lytA-encoded autolysin (N-acetylmuramoyl-L-alanine amidase) of Streptococcus pneumoniae is believed to play an important role in the pathogenesis of pneumococcal infection and has been identified as a putative vaccine target. Allelic diversity of lytA in an extensive collection of clinical isolates was assessed by restriction fragment length polymorphism and confirmatory sequencing studies. Genetic diversity within lytA is limited, especially compared to the high levels of diversity seen in other pneumococcal virulence factor genes, although small blocks generating mosaic structure were identified. Sequence comparisons with genes encoding cell wall lytic enzymes of pneumococcal bacteriophage suggest that localized recombination events have occurred between host lytA and these bacteriophage genes. These results confirm earlier suggestions that recombination between DNA encoding bacteriophage autolytic enzymes and chromosomally encoded lytA might be important in the evolution of lytA. The implications of these findings for understanding the evolution of lytA and the potential utility of LytA as a vaccine target are discussed. PMID- 10456898 TI - Role of tumor necrosis factor alpha, interleukin-1beta, and interleukin-6 in a mouse model of group B streptococcal arthritis. AB - Intravenous inoculation of CD1 mice with 10(7) CFU of type IV group B Streptococcus (GBS IV) results in a high incidence of diffuse septic arthritis. In this study the roles of tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), and IL-6 in articular pathology were evaluated. Cytokine levels were quantified in the serum and joints by enzyme-linked immunosorbent assay in mice injected with GBS IV and tested or not tested with pentoxifylline (PTF), a methylxanthine that affects cytokine production. PTF was administered intraperitoneally at a dose of 1 mg/mouse (50 mg/kg of body weight) 1 h after GBS infection and then at 24-h intervals for 4 days. High levels of IL-1beta and IL 6, but not TNF-alpha, were detected in the joints of mice injected with GBS IV from 5 to 15 days after infection, when articular lesions were most frequent and severe. IL-1beta and IL-6 concentrations in the joints significantly (P < 0.001) exceeded those detected in the serum, confirming a strong local production. PTF treatment resulted in a strong reduction of cytokine production and in a marked decrease in both the incidence and severity of arthritis. Inoculation of exogenous murine recombinant IL-1beta or IL-6 in mice treated with GBS IV plus PTF resulted in an incidence and severity of articular lesions similar to those obtained with inoculation of GBS IV alone. No significant effect was obtained with TNF-alpha administration. These data show a strong involvement of IL-1beta and IL-6, but not TNF-alpha, in the pathogenesis of GBS arthritis. PMID- 10456900 TI - Identification of a new variable sequence in the P1 cytadhesin gene of Mycoplasma pneumoniae: evidence for the generation of antigenic variation by DNA recombination between repetitive sequences. AB - A Mycoplasma pneumoniae cytadhesin P1 gene with novel nucleotide sequence variation has been identified. Four clinical strains of M. pneumoniae were found to carry this type of P1 gene. This new P1 gene is similar to the known group II P1 genes but possesses novel sequence variation of approximately 300 bp in the RepMP2/3 region. The position of the new variable region is distant from the previously reported variable regions known to differ between group I and II P1 genes. Two sequences closely homologous to this new variable region were found within the repetitive sequences outside the P1 gene of the M. pneumoniae M129 genome. This suggests that the new P1 gene was generated by DNA recombination between repetitive sequences and the P1 gene locus. The finding of this new type of P1 gene supports the hypothesis that the repetitive sequences of the M. pneumoniae genome serve as a reservoir to generate antigenic variation of the cytadhesin P1 gene. PMID- 10456901 TI - Binding of rat and human surfactant proteins A and D to Aspergillus fumigatus conidia. AB - Surfactant proteins A (SP-A) and D (SP-D) are thought to play important roles in pulmonary host defense. We investigated the interactions of rat and human SP-A and SP-D with Aspergillus fumigatus conidia. Rat SP-D but not rat SP-A bound the conidia, and the binding was inhibited by EDTA, mannose, glucose, maltose, and inositol. Binding studies using a mutant recombinant rat SP-D with altered carbohydrate recognition but normal structural organization clearly established a role for the carbohydrate recognition domain in binding to conidia. However, neither rat SP-A nor SP-D increased the association of fluorescein isothiocyanate labeled conidia with rat alveolar macrophages as determined by flow cytometry. Both human SP-A (isolated from normal and alveolar proteinosis lungs) and SP-D (recombinant protein and protein isolated from alveolar proteinosis lungs) bound the conidia. These data indicate that important differences exist between rat and human SP-A in binding to certain fungi. Human SP-A and SP-D binding to conidia was also examined in the presence of hydrophobic surfactant components (HSC), containing both the phospholipid and hydrophobic proteins of surfactant. We found that HSC inhibited but did not eliminate human SP-A binding to Aspergillus conidia. In contrast, the SP-D binding to conidia was unaffected by HSC. These findings indicate that SP-D plays a major role in the recognition of Aspergillus conidia in alveolar fluid. PMID- 10456902 TI - Polarization of the immune response to the single immunodominant epitope of p38, a major Schistosoma mansoni egg antigen, generates Th1- or Th2-type cytokines and granulomas. AB - In schistosomiasis mansoni, helminth eggs secrete soluble egg antigens (SEA) that induce T-cell-mediated granulomatous tissue responses. The cloned 38-kDa peptide (p38) of SEA was shown to induce and elicit Th1-type responsiveness in H-2(k) mice. Subsequently, the immunodominant T-cell epitope (P4) of p38 was shown to elicit pulmonary granuloma formation and Th1-type cytokine production in sensitized or infected mice. Here, we report that the immune response to p38 or P4 can be polarized to a Th1 or Th2 profile when the peptides are presented intraperitoneally in soluble recombinant interleukin-12 (IL-12) or alum adjuvant, respectively. The Th1 or Th2 profile was verified by cytokine secretion, enzyme linked spot assay, and antibody isotype characterization. Importantly, the polarized immune response generated two types of pulmonary granulomas around injected P4-coated beads. The type 1 granulomas were smaller and contained mononuclear cells and occasional thin strands of deposited collagen. In contrast, the type 2 lesions were larger and contained mononuclear cells, large numbers of eosinophils, and several thick bands of deposited collagen. By reverse transcription-PCR cytokine, message in the type 1 granuloma-bearing lungs was found for gamma interferon, tumor necrosis factor alpha, and inducible nitric oxide synthase but not for IL-4 or IL-5. Conversely, lungs with type 2 granulomas had message only for IL-4 and IL-5. These results show that in the proper cytokine environment, the response to a strong Th1 inducer peptide can be deviated to a Th2 profile. PMID- 10456903 TI - Analysis of antigenic structure and human immune response to outer membrane protein CD of Moraxella catarrhalis. AB - Moraxella catarrhalis is an important cause of otitis media in children and lower respiratory tract infections in adults with chronic obstructive pulmonary disease (COPD). Outer membrane protein CD (OMP CD) is a 45-kDa protein which is a potential vaccine antigen to prevent infections caused by M. catarrhalis. Eight monoclonal antibodies were used to study the antigenic structure of the OMP CD molecule by assaying recombinant peptides corresponding to the sequence of the protein. This approach identified two surface-exposed epitopes, including one near the amino terminus (amino acids 25 to 44) and one in the central region of the molecule (amino acids 261 to 331). Assays with serum and sputum supernatants of adults with COPD revealed variable levels of antibodies to OMP CD among individuals. To determine which portions of the OMP CD molecule were recognized by human antibodies, three human serum samples were studied with six recombinant peptides which span the sequence of OMP CD. All three sera contained immunoglobulin G antibodies which recognized exclusively the peptide corresponding to amino acids 203 to 260 by immunoblot assay. Adsorption experiments with whole bacteria established that some of the human antibodies are directed at surface-exposed epitopes on OMP CD. We conclude that OMP CD is a highly conserved molecule which contains at least two separate epitopes which are exposed on the bacterial surface. While individual adults with COPD show variability in the immune response to OMP CD, a specific region of the OMP CD molecule (amino acids 203 to 260) is important as a target of the human immune response. PMID- 10456904 TI - Assessment of immunity to mycobacterial infection with luciferase reporter constructs. AB - Protective immunity to mycobacterial infection is incompletely understood but probably involves the coordinated interaction of multiple cell types and cytokines. With the aim of developing assays that might provide a surrogate measure of protective immunity, we have investigated the use of recombinant mycobacteria carrying luciferase reporter enzymes to assess the effectiveness of antimycobacterial immunity in model systems. Measurement of luminescence was shown to provide a rapid and simple alternative to the counting of CFU as a means of monitoring mycobacterial viability. We describe optimization of a luciferase reporter strain of Mycobacterium tuberculosis and demonstrate its application for the study of mycobacterial interactions with host cells in tissue culture and the rapid assessment of vaccine efficacy in a murine model. PMID- 10456905 TI - Murine splenocytes induce severe gastritis and delayed-type hypersensitivity and suppress bacterial colonization in Helicobacter pylori-infected SCID mice. AB - The goal of this study was to evaluate the role of host immunity in gastritis and epithelial damage due to Helicobacter pylori. Splenocytes from H. pylori-infected and uninfected C57BL/6 mice were adoptively transferred to H. pylori-infected and uninfected severe combined immunodeficient (SCID) mice. Transfer was verified by flow cytometry, and all mice were evaluated for the presence of delayed-type hypersensitivity (DTH) by footpad inoculation with sterile H. pylori sonicate and for humoral immunity by enzyme-linked immunosorbent assay. The severity of gastritis and gastric epithelial damage was quantified histologically, epithelial proliferation was determined by proliferating cell nuclear antigen staining, and colonization was quantified by culture. C57BL/6 mice, but not nonrecipient SCID mice, developed moderate gastritis in response to H. pylori. In contrast, recipient SCID mice developed severe gastritis involving 50 to 100% of the gastric mucosa and strong DTH responses not present in C57BL/6 mice. DTH, but not serum anti-H. pylori immunoglobulin G, correlated with adoptive transfer, gastritis, and bacterial clearance. Severe gastritis, but not bacterial colonization, was associated with epithelial metaplasia, erosions, and an elevated labeling index. This study demonstrates that (i) adaptive immunity is essential for development of gastritis due to H. pylori in mice, (ii) T-cell enriched lymphocytes in SCID mice induce DTH and gastritis, which is more severe than donor gastritis, and (iii) the host inflammatory response, not direct bacterial contact, causes epithelial damage. The greater severity of gastritis in recipient SCID mice than in donor C57BL/6 mice suggests that gastritis is due to specific T-cell subsets and/or the absence of regulatory cell subsets in the transferred splenocytes. PMID- 10456906 TI - Immunization of mice with a TolA-like surface protein of Trypanosoma cruzi generates CD4(+) T-cell-dependent parasiticidal activity. AB - The gene family encoding a trypomastigote-specific protein restricted to the part of the flagellum in contact with the cell body of the trypomastigote form of Trypanosoma cruzi has been isolated, characterized, and expressed in a baculovirus expression system. The gene family contains three tandemly repeated members that have 97 to 100% sequence identity. The predicted protein encoded by the gene family has both significant amino acid sequence identity and other physical and biological features in common with the TolA proteins of Escherichia coli and Pseudomonas aeruginosa. Based on these similarities, we have designated this gene family tolT. Immunization of mice with recombinant TolT generates a population of CD4(+) T lymphocytes that recognize T. cruzi-infected macrophages, resulting in the production of gamma interferon (IFN-gamma), which leads to NO production and a 50 to 60% reduction in parasite numbers compared to that seen with infected macrophages incubated with naive T cells. This population of T cells also produces both IFN-gamma and interleukin 2 (IL-2) but not IL-4 or IL-5 when incubated with spleen cells stimulated with TolT antigen, indicating that they are of the T-helper 1 type. T cells from mice chronically infected with T. cruzi also produce significant levels of IFN-gamma when cocultured with macrophages and either TolT protein or paraflagellar rod protein, indicating that both of these flagellar proteins produce positive T-cell responses in mice chronically infected with T. cruzi. PMID- 10456907 TI - Pseudomonas aeruginosa exoenzyme S stimulates murine lymphocyte proliferation in vitro. AB - The exuberant immunoinflammatory response that is associated with Pseudomonas aeruginosa infection is the major source of the morbidity and mortality in cystic fibrosis (CF) patients. Previous studies have established that an exoproduct of P. aeruginosa (exoenzyme S) is a mitogen for human T lymphocytes and activates a larger percentage of T cells than most superantigens, which may contribute to the immunoinflammatory response. An animal model would facilitate studies of the pathophysiologic consequences of this activation. As a first step toward developing an animal model, the murine lymphocyte response to exoenzyme S was examined. When stimulated with exoenzyme S, splenocytes isolated from naive mice entered S phase and proliferated. The optimum response occurred after 2 to 3 days in culture, at 4 x 10(5) cells per well and 5.0 micrograms of exoenzyme S per ml. The response was not due to lipopolysaccharide, since Rhodobacter sphaeroides lipid A antagonist did not block the response. Other preparations of exoenzyme S stimulated lymphocyte proliferation, since the response to recombinant exoenzyme S (rHisExo S) cloned from strain 388 was similar to the response to exoenzyme S from strain DG1. There was evidence that genetic variability influenced the response, since A/J, CBA/J, and C57BL/6 mice were high responders and BALB/cJ mice were low responders following stimulation with exoenzyme S. Both splenic T and B lymphocytes entered the cell cycle in response to exoenzyme S. Thus, murine lymphocytes, like human lymphocytes, respond to P. aeruginosa exoenzyme S, which supports the development of a murine model that may facilitate our understanding of the role that exoenzyme S plays in the pathogenesis of P. aeruginosa infections in CF patients. PMID- 10456908 TI - The capsule of Cryptococcus neoformans reduces T-lymphocyte proliferation by reducing phagocytosis, which can be restored with anticapsular antibody. AB - Cell-mediated immunity is critical for the host defense to Cryptococcus neoformans, as demonstrated by numerous animal studies and the prevalence of the infection in AIDS patients. Previous studies have established that the polysaccharide capsule contributes to the virulence of C. neoformans by suppressing T-lymphocyte proliferation, which reflects the clonal expansion of T lymphocytes that is a hallmark of cell-mediated immunity. The present studies were performed to identify the major mechanism by which polysaccharide impairs lymphocyte proliferation, since capsular polysaccharide has the potential to affect the development of T-lymphocyte responses by stimulating production of interleukin-10 (IL-10), inhibiting phagocytosis, and inducing shedding of cell surface receptors. We demonstrate that polysaccharide inhibits lymphocyte proliferation predominantly by blocking uptake of C. neoformans, which is crucial for subsequent lymphocyte proliferation. In addition, we show that polysaccharide did not suppress lymphocyte proliferation via an IL-10-dependent mechanism, nor did it affect critical surface receptor interactions on the T cell or antigen presenting cell. Having established that polysaccharide impairs phagocytosis, we performed studies to determine whether opsonization with human serum or with anticapsular antibody could reverse this effect. Impaired uptake and lymphocyte proliferation that were induced by polysaccharide can be enhanced through opsonization with monoclonal antibodies or human serum, suggesting that antipolysaccharide antibodies might enhance the host defense by restoring uptake of the organism and subsequent presentation to T lymphocytes. These studies support the therapeutic potential of stimulating cell-mediated immunity to C. neoformans with anticapsular antibody. PMID- 10456909 TI - Attenuation and immunogenicity of Deltacya Deltacrp derivatives of Salmonella choleraesuis in pigs. AB - Six different isogenic Deltacya Deltacrp derivatives of a strain of Salmonella choleraesuis var. kunzendorf-chi3246 virulent for pigs were constructed by transposon-mediated deletion mutagenesis. These strains were evaluated for virulence and ability to elicit a protective immune response in young weaned pigs after oral administration and were compared to a commercially available vaccine which lacks the 50-kb virulence plasmid (vpl(-)). These derivatives were Deltacya Deltacrp vpl(+), Deltacya Deltacrp vpl(-), Deltacya Delta(crp-cdt) vpl(+), Deltacya Delta(crp-cdt) vpl(-), Deltacya Deltacrp pmi-3834 vpl(+), and Deltacya Delta(crp-cdt) pmi-3834. In experiments to evaluate safety, no significant adverse effects of any of the vaccine constructs were observed, except that two of the strains which carried the virulence plasmid (vpl(+)) caused a small, short term elevation in maximum temperature compared to pretreatment temperature values. Orally immunized animals, except for those vaccinated with the Deltacya Deltacrp pmi-3834 vpl(+) strain or SC-54, developed significant serum antibody responses 21 days postvaccination as measured by enzyme-linked immunosorbent assay. No cell-mediated immune responses to heat-killed S. choleraesuis were noted at the same time point as measured with heat-killed bacteria as antigen in a lymphocyte proliferation assay. In an oral challenge exposure model with a highly virulent heterologous strain of S. choleraesuis, the Deltacya Deltacrp strains with deletions in pmi were not protective. As measured by morbidity scores, the responses to challenge of the pigs vaccinated with the other four Deltacya Deltacrp derivatives were significantly better than those of the nonvaccinated, challenged group. With the exception of temperature elevation and slight differences in diarrhea scores postchallenge, none of these strains differed significantly from each other in the other clinical parameters analyzed. While the commercial vaccine was protective by most of the parameters measured, it was not fully protective against challenge with virulent S. choleraesuis as judged by diarrhea scores and temperature elevation. Collectively, these data demonstrate that Deltacya Deltacrp derivatives, with or without the virulence plasmid but not with deletions in the pmi gene, are candidates for vaccines for protection against salmonellosis in pigs. PMID- 10456910 TI - Characterization of VspB of Borrelia turicatae, a major outer membrane protein expressed in blood and tissues of mice. AB - Serotypes A and B of the relapsing fever spirochete Borrelia turicatae produce different disease manifestations in infected mice. Whereas serotype B causes more severe arthritis and reaches higher densities in the blood of mice than serotype A, serotype A invades the central nervous system earlier than serotype B during infection. These differences between serotypes A and B in mice are associated with the expression of different surface proteins, VspA and VspB, respectively, in the culture medium. To determine whether these proteins, in particular, VspB, are also expressed in vivo, scid mice infected with B. turicatae were studied. The expression of VspB by spirochetes in the blood was demonstrated in Coomassie blue-stained polyacrylamide gels and Western blots with a specific monoclonal antibody. Indirect immunofluorescence and immunoperoxidase studies confirmed the expression of VspB in the blood and also demonstrated VspB expression in the joints and heart. The gene for VspB was next identified and cloned by using partial amino acid sequencing, reverse transcriptase PCR, and a specific monoclonal antibody. The vspB gene encodes a protein of 216 amino acids that is 68% identical to VspA of B. turicatae and 44 to 56% identical to representative Vsp and OspC lipoproteins of other Borrelia spp. The processed VspB protein was distinguished from 26 other Vsp and OspC proteins by a high predicted isoelectric point at 9.39. The promoter region for vspB was similar to the promoter region for the vsp33 gene of Borrelia hermsii and for the ospC gene of Borrelia burgdorferi, two genes known to be environmentally regulated. These studies established that the virulence-associated VspB protein is expressed by spirochetes in the mouse and that VspB is a novel member of the Vsp-OspC family of proteins. PMID- 10456911 TI - Heat-killed Streptococcus suis capsular type 2 strains stimulate tumor necrosis factor alpha and interleukin-6 production by murine macrophages. AB - Streptococcus suis capsular type 2 is an important etiological agent of swine meningitis, and it is also a zoonotic agent. Since mononuclear phagocytes have been suggested to play a central role in the pathogenesis of meningitis, the objective of the present study was to evaluate the capacity of whole killed S. suis type 2 organisms to induce the release of the proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) by murine macrophages. Induction of cytokines was evaluated in the presence or absence of phorbol ester (phorbol 12-myristate 13-acetate [PMA]) costimulation. Results showed that S. suis type 2 stimulated the production of both cytokines in a concentration- and time-dependent fashion. Although large doses of bacteria were required for maximal cytokine release, titers were similar to those obtained with the lipopolysaccharide (LPS) positive control. An increase in cytokine release was observed with both S. suis and LPS with PMA costimulation. Experiments with cytochalasin-treated macrophages showed that the stimulation of cytokine production was phagocytosis independent. When macrophages were stimulated with an unencapsulated mutant, an increase in TNF production was observed, but the absence of the capsule had no effect on IL-6 production. In fact, whereas purified capsular polysaccharide of S. suis failed to induce cytokine release, purified S. suis cell wall induced both TNF and, to a lesser extent, IL-6. IL-6 secretion probably requires some distinct stimuli which differ from those of TNF. Finally, the S. suis putative virulence factors suilysin and extracellular protein EF showed no cytokine-stimulating activity. The ability of S. suis to trigger macrophages to produce proinflammatory cytokines may have an important role in the initiation and development of meningitis caused by this microorganism. PMID- 10456912 TI - Control of white-opaque phenotypic switching in Candida albicans by the Efg1p morphogenetic regulator. AB - Phenotypic switching in Candida albicans spontaneously generates different cellular morphologies and is manifested in strain WO-1 by the reversible switching between the white and opaque phenotypes. We present evidence that phenotypic switching is regulated by the Efg1 protein, which is known as an essential element of hyphal development (dimorphism). Firstly, EFG1 is expressed specifically in cells of the white but not the opaque phenotype. During mass conversion from the opaque to the white phenotype, the EFG1 transcript level correlates with competence of switching of opaque cells to the white form. Secondly, overexpression of EFG1 by a PCK1p-EFG1 fusion forces opaque-phase cells to switch to the white form with a high level of efficiency. Thirdly, low-level expression of EFG1 in strain CAI-8 generates a cellular phenotype similar to that of opaque cells in that cells bud as short rods, which cannot be induced to form hyphae in standard conditions; such cells (unlike authentic opaque cells) lack typical surface "pimples." Importantly, the opaque-specific OP4 transcript is induced in the opaque-like cells generated by strain CAI8 as a response to low level expression of EFG1. The results suggest that high EFG1 expression levels induce and maintain the white cell form while low EFG1 expression levels induce and maintain the opaque cell form. It is proposed that changes in EFG1 expression determine or contribute to phenotypic switching events in C. albicans. PMID- 10456913 TI - Dominance of immunoglobulin G2c in the antiphosphorylcholine response of rats infected with Trichinella spiralis. AB - The antibody response to the L1 stage of Trichinella spiralis has been described as biphasic. Worms resident in the intestine during the first week of infection stimulate an antibody response against a subset of larval proteins. L1 larvae in the muscle at the end stage of infection stimulate a second antibody response against tyvelose-bearing glycoproteins. Antityvelose antibodies protect rats against challenge infection with larvae. The aim of this study was to characterize the rat B-cell response against larval antigens during the intestinal phase of T. spiralis infection and to test the antiparasitic effects of such antibodies. Strain PVG rats were infected orally with 500 larvae. Antibodies specific for phosphorylcholine-bearing proteins of L1 larvae first appeared in serum 9 days postinfection. Absorption experiments showed that the majority of antilarval antibodies produced in rats 16 days after infection with T. spiralis were specific for phosphorylcholine-bearing proteins. A fraction of these antibodies bound to free phosphorylcholine. Immunoglobulin G2c (IgG2c) producing cells in the mesenteric lymph node dominated this early antibody response. IgG2c is associated with T-independent immune responses in the rat; however, a comparison of athymic rats with euthymic controls suggested that only a small fraction of the phosphorylcholine-related antibody response against T. spiralis was T independent. Phosphorylcholine is a common epitope in antigens of bacteria and nematode parasites and has been shown to be a target of protective immunity in certain bacteria. A monoclonal IgG2c antibody was prepared from infected rats and shown to be specific for phosphorylcholine. Monoclonal phosphorylcholine-specific IgG2c failed to protect rats against intestinal infection with T. spiralis. Therefore, our findings do not support a role for phosphorylcholine-bearing antigens in immune defense against T. spiralis; however, the potency of the immune response induced suggests an immunomodulatory role for the lymphocytes involved. PMID- 10456914 TI - Lactoferrin-lipid A-lipopolysaccharide interaction: inhibition by anti-human lactoferrin monoclonal antibody AGM 10.14. AB - Lactoferrin (LF) is a glycoprotein that exerts both bacteriostatic and bactericidal activities. The interaction of LF with lipopolysaccharide (LPS) of gram-negative bacteria seems to play a crucial role in the bactericidal effect. In this study, we evaluated, by means of an enzyme-linked immunosorbent assay, the binding of biotinylated LF to the S (smooth) and R (rough) (Ra, Rb, Rc, Rd1, Rd2, and Re) forms of LPS and different lipid A preparations. In addition, the effects of two monoclonal antibodies (AGM 10.14, an immunoglobulin G1 [IgG1] antibody, and AGM 2.29, an IgG2b antibody), directed against spatially distant epitopes of human LF, on the LF-lipid A or LF-LPS interaction were evaluated. The results showed that biotinylated LF specifically binds to solid-phase lipid A, as this interaction was prevented in a dose-dependent fashion by either soluble uncoupled LF or lipid A. The binding of LF to S-form LPS was markedly weaker than that to lipid A. Moreover, the rate of LF binding to R-form LPS was inversely related to core length. The results suggest that the polysaccharide O chain as well as oligosaccharide core structures may interfere with the LF-lipid A interaction. In addition, we found that soluble lipid A also inhibited LF binding to immobilized LPS, demonstrating that, in the whole LPS structure, the lipid A region contains the major determinant recognized by LF. AGM 10.14 inhibited LF binding to lipid A and LPS in a dose-dependent fashion, indicating that this monoclonal antibody recognizes an epitope involved in the binding of LF to lipid A or some epitope in its close vicinity. In contrast, AGM 2.29, even in a molar excess, did not prevent the binding of LF to lipid A or LPS. Therefore, AGM 10.14 may represent a useful tool for neutralizing selectively the binding of LF to lipid A. In addition, the use of such a monoclonal antibody could allow better elucidation of the consequences of the LF-lipid A interaction. PMID- 10456915 TI - Fibronectin binding protein and host cell tyrosine kinase are required for internalization of Staphylococcus aureus by epithelial cells. AB - Staphylococcus aureus expresses several surface proteins that promote adherence to host cell extracellular matrix proteins, including fibronectin (Fn). Since this organism has recently been shown to be internalized by nonprofessional phagocytes, a process that typically requires high-affinity binding to host cell receptors, we investigated the role of its Fn binding proteins (FnBPs) and other surface proteins in internalization by the bovine mammary gland epithelial cell line (MAC-T). Efficient internalization of S. aureus 8325-4 required expression of FnBPs; an isogenic mutant (DU5883), not expressing FnBPs, was reduced by more than 95% in its ability to invade MAC-T cells. Moreover, D3, a synthetic peptide derived from the ligand binding domain of FnBP, inhibited the internalization of the 8325-4 strain in a dose-dependent fashion and the efficiency of staphylococcal internalization was partially correlated with Fn binding ability. Interestingly, Fn also inhibited the internalization and adherence of S. aureus 8325-4 in a dose-dependent manner. In contrast to internalization, adherence of DU5883 to MAC-T was reduced by only approximately 40%, suggesting that surface binding proteins, other than FnBPs, can mediate bacterial adherence to cells. Adherence via these proteins, however, does not necessarily result in internalization of the staphylococci. An inhibitor of protein tyrosine kinase, genistein, reduced MAC-T internalization of S. aureus by 95%, indicating a requirement for a host signal transduction system in this process. Taken together, these results indicate that S. aureus invades nonprofessional phagocytes by a mechanism requiring interaction between FnBP and the host cell, leading to signal transduction and subsequent rearrangement of the host cell cytoskeleton. PMID- 10456916 TI - Isolation of peptides that mimic epitopes on a malarial antigen from random peptide libraries displayed on phage. AB - The ring-infected erythrocyte surface antigen (RESA) is a dense-granule protein of Plasmodium falciparum which binds to the cytoskeletal structure of the erythrocyte after parasite invasion. It is currently under trial as a vaccine candidate. In an effort to characterize further the antibody responses to this antigen, we have panned two independent libraries of random peptides expressed on the surface of filamentous phage with a monoclonal antibody (MAb 18/2) against RESA. One library consisted of a potentially constrained 17-mer peptide fused with the gpVIII phage coat protein, and the other displayed an unconstrained 15 mer as a fusion with the minor phage coat protein gpIII. Several rounds of biopanning resulted in enrichment from both libraries clones that interacted specifically with MAb 18/2 in protein-blotting and enzyme-linked immunosorbent assay experiments. Nucleotide sequencing of the random oligonucleotide insert revealed a common predominant motif: (S/T)AVDD. Several other clones had related but degenerate motifs. Thus, a monoclonal antibody against a malarial antigen can select common mimotopes from different random peptide libraries. We envisage many uses for this technology in malaria research. PMID- 10456917 TI - Genetic control of schistosome infections by the SM1 locus of the 5q31-q33 region is linked to differentiation of type 2 helper T lymphocytes. AB - Human susceptibility to Schistosoma mansoni infections is controlled by the SM1 locus on chromosome 5 in q31-q33. This genetic region encodes cytokines which regulate the development of helper T lymphocytes. In the present work, a clonal analysis of CD4(+) T lymphocytes of homozygous resistant and homozygous susceptible subjects was undertaken to evaluate whether SM1 controls helper T cell differentiation. Of 121 CD4(+) T-cell clones (TCC) from three susceptible (S) and three resistant (R) subjects, 68 proliferated when stimulated by parasite antigens. Parasite-specific TCC derived from susceptible subjects (33 STCC) produced 10- to 1,000-fold less interleukin-4 and -5 than TCC from resistant subjects (25 RTCC). Clones from both patient groups produced, however, the same amount of gamma interferon. Parasite-specific STCC were type 1 helper (Th1) or Th0/1, whereas RTCC were either Th2 or Th0/2. These results, together with the localization of SM1 in 5q31-q33, indicate that the SM1 locus controls the differentiation of Th2 lymphocytes. PMID- 10456918 TI - Regulation of protein phosphorylation and pathogen phagocytosis by surfactant protein A. AB - Surfactant protein A (SP-A), a pulmonary member of the collectin family of proteins, facilitates the rapid clearance of pathogens by upregulating immune cell functions in the lungs. SP-A binds to bacteria and targets them for rapid phagocytosis by alveolar macrophages, but the mechanism by which this stimulation occurs is not clear. To characterize the intracellular events that may be involved, we examined the roles of protein phosphorylation and cytoskeletal polymerization in SP-A-stimulated phagocytosis. In rat alveolar macrophages, SP-A stimulated rapid tyrosine phosphorylation of specific proteins in a dose- and time-dependent manner. The pattern of proteins that were phosphorylated in response to SP-A, as resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was similar to that observed for immunoglobulin G (IgG) stimulated macrophages. Both SP-A and IgG stimulated increases in phagocytosis of Streptococcus pneumoniae above levels in the absence of added protein by 394% +/- 81% and 200% +/- 25%, respectively. Phagocytosis in both cases was dependent on tyrosine kinases, protein kinase C, and actin polymerization but not on microtubule activity. These studies show that SP-A utilizes pathways similar to those used by IgG to increase macrophage phagocytosis of bacteria. PMID- 10456919 TI - Preliminary characterization of a Mycobacterium abscessus mutant in human and murine models of infection. AB - The ability to persist in the host after the establishment of infection is an important virulence determinant for mycobacteria. Mycobacterium abscessus is a rapidly growing mycobacterial species which causes a variety of clinical syndromes in humans. We have obtained a rough, wild-type human clinical isolate of M. abscessus (M. abscessus-R) and a smooth, attenuated mutant (M. abscessus-S) which spontaneously dissociated from the clinical isolate. We have found that M. abscessus-R is able to persist and multiply in a murine pulmonary infection model in contrast to M. abscessus-S, which is rapidly cleared. To understand the basis for this difference, we characterized the behavior of these variants in human tissue culture models of infection. M. abscessus-R is able to persist and multiply in human monocytes, while M. abscessus-S is deficient in this ability. Both of these variants are phagocytized by human monocytes. M. abscessus-R resides in a phagosome typical for pathogenic mycobacteria with a tightly adherent phagosomal membrane. In contrast, M. abscessus-S resides in a "loose" phagosome with the phagosomal membrane separated from the bacterial cell wall. Both M. abscessus variants also have distinctive growth patterns in a recently described fibroblast-mycobacterium microcolony assay, with M. abscessus-R exhibiting growth characteristics similar to those previously reported for virulent M. tuberculosis and M. abscessus-S exhibiting growth characteristics similar to those previously reported for avirulent M. tuberculosis. In both the monocyte infection assay and the murine pulmonary infection model, numerous infected mononuclear phagocyte aggregates develop at sites of M. abscessus-R infection, but are absent with M. abscessus-S infection. We conclude that a mutation has occurred in the M. abscessus-S variant which has altered the ability of this organism to persist and multiply in host cells and that this may be related to the phenotypic changes we have observed in our tissue culture models of infection. PMID- 10456920 TI - Pure botulinum neurotoxin is absorbed from the stomach and small intestine and produces peripheral neuromuscular blockade. AB - Clostridium botulinum serotype A produces a neurotoxin composed of a 100-kDa heavy chain and a 50-kDa light chain linked by a disulfide bond. This neurotoxin is part of a ca. 900-kDa complex, formed by noncovalent association with a single nontoxin, nonhemagglutinin subunit and a family of hemagglutinating proteins. Previous work has suggested, although never conclusively demonstrated, that neurotoxin alone cannot survive passage through the stomach and/or cannot be absorbed from the gut without the involvement of auxiliary proteins in the complex. Therefore, this study compared the relative absorption and toxicity of three preparations of neurotoxin in an in vivo mouse model. Equimolar amounts of serotype A complex with hemagglutinins, complex without hemagglutinins, and purified neurotoxin were surgically introduced into the stomach or into the small intestine. In some experiments, movement of neurotoxin from the site of administration was restricted by ligation of the pylorus. Comparison of relative toxicities demonstrated that at adequate doses, complex with hemagglutinins, complex without hemagglutinins, and pure neurotoxin can be absorbed from the stomach. The potency of neurotoxin in complex was greater than that of pure neurotoxin, but the magnitude of this difference diminished as the dosage of neurotoxin increased. Qualitatively similar results were obtained when complex with hemagglutinins, complex without hemagglutinins, and pure neurotoxin were placed directly into the intestine. This work establishes that pure botulinum neurotoxin serotype A is toxic when administered orally. This means that pure neurotoxin does not require hemagglutinins or other auxiliary proteins for absorption from the gastrointestinal system into the general circulation. PMID- 10456921 TI - Isolation of recombinant protective Helicobacter pylori antigens. AB - A total of seven clones producing both new and previously described Helicobacter pylori proteins were isolated from a library of H. pylori genomic DNA. The screening approach by which these proteins were detected relied on the use of antisera raised in mice vaccinated with Helicobacter felis sonicate plus cholera toxin, a regimen which protects mice from H. pylori challenge. This strategy was designed to maximize the possibility of obtaining antigens which might be capable of conferring protection from H. pylori infection. Two of the clones were shown to encode the urease enzyme and the heat shock protein HspB, which have already been identified as protective antigens. The other five clones were sequenced, protein coding regions were deduced, and these sequences were amplified by PCR for incorporation into Escherichia coli expression vectors. The proteins produced from these expression systems were purified to allow testing for protective efficacy in an H. pylori mouse model. All five proteins were able to facilitate the clearance of a challenge with H. pylori, as judged by an assay of gastric urease activity and light microscopy on stomach sections. These results clearly indicate that the screening strategy has successfully identified candidate vaccine antigens. PMID- 10456922 TI - Pneumococcal surface protein A inhibits complement activation by Streptococcus pneumoniae. AB - Pneumococcal surface protein A (PspA) is a surface-exposed protein virulence factor for Streptococcus pneumoniae. In this study, no significant depletion of serum complement was observed for the serum of mice infected with pneumococci that express PspA. In contrast, in mice infected with an isogenic strain of pneumococci lacking PspA, significant activation of serum complement was detected within 30 min after infection. Also, the PspA-deficient strain but not the PspA expressing strain was cleared from the blood within 6 h. The contribution of PspA to pneumococcal virulence was further investigated by using mice deficient for C5, C3, or factor B. In mice deficient for C3 or factor B, PspA-negative pneumococci became fully virulent. In contrast, in C5-deficient mice as in wild type mice, PspA-deficient pneumococci were avirulent. These in vivo data suggest that, in nonimmune mice infected with pneumococci, PspA interferes with complement-dependent host defense mechanisms mediated by factor B. Immunoblots of pneumococci opsonized in vitro suggested that more C3b was deposited on PspA negative than on PspA-positive pneumococci. This was observed with and without anticapsular antibody. Furthermore, processing of the alpha chain of C3b was reduced in the presence of PspA. We propose that PspA exerts its virulence function by interfering with deposition of C3b onto pneumococci and/or by inhibiting formation of a fully functional alternative pathway C3 convertase. By blocking recruitment of the alternative pathway, PspA reduces the amount of C3b deposited onto pneumococci, thereby reducing the effectiveness of complement receptor-mediated pathways of clearance. PMID- 10456923 TI - Identification, cloning, and expression of the CAMP factor gene (cfa) of group A streptococci. AB - The CAMP reaction is a synergistic lysis of erythrocytes by the interaction of an extracellular protein (CAMP factor) produced by some streptococcal species with the Staphylococcus aureus sphingomyelinase C (beta-toxin). Group A streptococci (GAS [Streptococcus pyogenes]) have been long considered CAMP negative, and this reaction commonly has been used to distinguish GAS from Streptococcus agalactiae. We here provide evidence that GAS possess this gene and produce an extracellular CAMP factor capable of participating in a positive CAMP reaction. The S. pyogenes CAMP factor is specified by a 774-bp open reading frame homologous to the CAMP factor genes from S. agalactiae and Streptococcus uberis. This gene, designated cfa, was isolated on a 1,256-bp fragment and cloned in Escherichia coli. Recombinant clones of E. coli expressing cfa secreted an active CAMP factor. The deduced 28.5-kDa protein encoded by cfa consists of 257 amino acids, with a predicted 28-amino-acid signal peptide. The cfa gene is widely spread among GAS: 82 of 100 clinical GAS isolates produced a positive CAMP reaction. Of the CAMP negative strains, 17 of the 18 GAS strains contained the cfa gene. Additionally, CAMP activity was detected in streptococci from serogroups C, M, P, R, and U. The cfa gene was cloned and actively expressed in Escherichia coli and gene fusions were made, placing the beta-galactosidase gene (lacZ) under control of the cfa promoter. These cfa promoter-lacZ fusions were introduced into S. pyogenes via a bacteriophage-derived site-specific integration vector where they showed that the cfa gene has a strong promoter that may be subject to as-yet-unidentified regulatory factors. The results presented here, along with previous reports, indicate that the CAMP factor gene is fairly widespread among streptococci, being present at least in groups A, B, C, G, M, P, R, and U. PMID- 10456924 TI - Interactions of Penicillium marneffei with human leukocytes in vitro. AB - Penicillium marneffei, a dimorphic fungus endemic in parts of Asia, causes disease in those with impaired cell-mediated immunity, especially persons with AIDS. The histopathology of penicilliosis marneffei features the intracellular infection of macrophages. We studied the interactions between human leukocytes and heat-killed yeast-phase P. marneffei. Monocyte-derived macrophages bound and internalized P. marneffei in the presence of complement-sufficient pooled human serum (PHS). Binding and phagocytosis were still seen if PHS was heat inactivated or omitted altogether. The binding of unopsonized P. marneffei to monocyte derived macrophages occurred in the absence of divalent cations and was not affected by inhibitors of mannose and beta-glucan receptors or monoclonal antibodies directed against CD14 and CD11/CD18. Binding was profoundly inhibited by wheat germ agglutinin. A vigorous respiratory burst was seen in peripheral blood mononuclear cells (PBMC) stimulated with P. marneffei, regardless of whether the fungi were opsonized. However, tumor necrosis factor alpha (TNF alpha) release from PBMC stimulated with P. marneffei occurred only if serum was present. These data demonstrate that (i) monocyte-derived macrophages bind and phagocytose P. marneffei even in the absence of opsonization, (ii) binding is divalent cation independent but is inhibited by wheat germ agglutinin, suggesting that the major receptor(s) recognizing P. marneffei is a glycoprotein with exposed N-acetyl-beta-D-glucosaminyl groups, (iii) P. marneffei stimulates the respiratory burst regardless of whether opsonins are present, and (iv) serum factors are required for P. marneffei to stimulate TNF-alpha release. The ability of unopsonized P. marneffei to parasitize mononuclear phagocytes without stimulating the production of TNF-alpha may be critical for the virulence of this intracellular parasite. PMID- 10456925 TI - Staphylococcus aureus isolates from patients with Kawasaki disease express high levels of protein A. AB - Kawasaki disease (KD) is an acute vasculitis of young children that can be complicated by coronary artery abnormalities. Recent findings suggest that a superantigen(s) may play an important role in stimulating the immune activation associated with the disease, although the origin of this superantigen(s) is unclear. Staphylococcus aureus, isolated from the rectum or pharynx of patients with KD, secretes toxic shock syndrome toxin 1 (TSST-1). The KD isolates express low levels of other exoproteins compared to isolates from patients with toxic shock syndrome (TSS). Thus, it was previously suggested that the KD isolates may be defective in the global regulatory locus agr (for accessory gene regulator), which positively regulates these factors (D. Y. M. Leung et al., Lancet 342:1385 1388, 1993). Here we describe another characteristic of KD isolates. When considered collectively, the KD isolates were found to express higher levels of staphylococcal protein A than the TSS isolates, another characteristic of an agr defective phenotype. This correlated with a higher level of spa mRNA in these isolates. In contrast, the KD and TSS isolates expressed comparable levels of TSST-1, consistent with previous findings (D. Y. M. Leung et al., Lancet 342:1385 1388, 1993). Analysis of RNAIII transcript levels and nucleotide sequence analysis of the RNAIII-coding region suggested that the KD isolates are not defective in RNAIII, the effector molecule of the agr regulatory system. However, induction of RNAIII transcription in the KD isolates did not result in a dramatic decrease in the amount of spa mRNA, as has been reported for other strains (F. Vandenesch, J. Kornblum, and R. P. Novick, J. Bacteriol. 173:6313-6320, 1991). PMID- 10456926 TI - Pulmonary outcome in cystic fibrosis is influenced primarily by mucoid Pseudomonas aeruginosa infection and immune status and only modestly by genotype. AB - Whether allelic variants of the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) independently contribute to pulmonary outcome in CF patients has not been resolved. We used both cross-sectional and mixed-model longitudinal analyses of data from CF patients that were at least 12 years old to determine the influence on pulmonary function (percent predicted forced expiratory volume [FEV(1)]) of the CFTR gene genotype, gender, mucoid Pseudomonas aeruginosa (MPA) infection status, presence of total opsonic antibody to MPA, and, separately, the opsonic antibody activity specific to the mucoid exopolysaccharide (MEP) surface antigen. Two different factors were independently associated with the lack of MPA infection: a high level of MEP-specific opsonic activity (MSOA), implicating an immunologically based mechanism of resistance to infection, and a lack of any type of opsonic antibody to MPA, indicative of no significant exposure or infection. This latter phenotype was found in a subset of CF patients who carried at least one uncommon CFTR gene allele suggestive of a genetic basis for resistance to infection in this group of older CF patients. For CF patients in whom both CFTR gene alleles were identified by screening for the 12 most common variants (75% of alleles), cross-sectional analysis showed that MPA infection was best correlated with lower percent predicted FEV(1), while genotype (two versus one DeltaF508 CFTR gene allele) and a low level of MSOA were associated with increased risk of infection. A mixed-model analysis of longitudinal spirometric measurements that considered multiple risk factors to derive regression equations was used to determine which clinical parameters had the greatest effect on the annual rate of decline in percent predicted FEV(1). This analysis showed that the CFTR gene genotype only modestly modified the constant (y intercept) of the derived equations, while gender and MPA infection status had the largest effects on annual rates of decline in percent predicted FEV(1). These results indicate that the CFTR genotype is usually not a primary determinant of pulmonary function in most CF patients, but gender and MPA infection status are. Infection status is potentially influenced by both immunologic (a high level of MSOA) and genetic factors, such as carriage of a CFTR gene allele that leads to a diagnosis of CF but still confers resistance to infection that is comparable to that of the wild type CFTR gene. PMID- 10456927 TI - The gene locus yijP contributes to Escherichia coli K1 invasion of brain microvascular endothelial cells. AB - Most cases of Escherichia coli meningitis develop as a result of hematogenous spread, but it is not clear how circulating E. coli crosses the blood-brain barrier. A TnphoA mutant of E. coli K1 RS218 was shown to be significantly less invasive than its parent strain in bovine and human brain microvascular endothelial cells (BMEC), which constitute the blood-brain barrier. More importantly, traversal of the blood-brain barrier was significantly less with this mutant than with the parent strain in newborn rats with experimental hematogenous meningitis. A DNA segment containing the TnphoA insertion site was cloned from RS218, and the cloned DNA complemented the TnphoA mutant in invasion of BMEC. Nucleotide sequence revealed a near identity to that of a hypothetical yijP gene (also called f577) in the E. coli K-12 genome. Sequence analysis indicated that the E. coli K1 yijP gene likely encodes a 66. 6-kDa membrane protein. Deletion and complementation experiments indicated that the yijP gene was involved in E. coli K1 invasion of BMEC, i.e., the invasive ability of E. coli K1 was significantly reduced after yijP was deleted and was restored by complementation with a plasmid containing the yijP open reading frame. This is the first demonstration that the yijP gene locus plays a role in the pathogenesis of E. coli K1 meningitis. PMID- 10456928 TI - T-Cell responses to Treponema pallidum subsp. pallidum antigens during the course of experimental syphilis infection. AB - In this study we describe the development of the T-cell response to a panel of Treponema pallidum antigens over the course of syphilis infection in the rabbit and determine whether these antigens induce the expression of Th1 cytokines. It was determined that the membrane proteins TpN17 and TpN47, as well as the endoflagellar sheath protein TpN37, induce strong proliferation responses through most of syphilis infection; Tromp1 induced only weak proliferative responses. An unexpected drop in proliferative response to these antigens at day 90 of infection, followed by a dramatic increase in response at day 180, suggests that there may be a secondary dissemination of T. pallidum which induces a recall response. Crude epitope mapping of TpN17 and TpN37 showed that multiple epitopes may be present on both antigens, which is likely a contributing factor in the immunodominance of these antigens. The T-cell response to the TpN37 molecule shows acquisition of newly recognized epitopes during the course of infection. Sonicated T. pallidum was found to induce the expression of interleukin-2 (IL-2) and gamma interferon and not IL-10 mRNA, showing that the general T-cell response to T. pallidum antigens in syphilis infection is biased towards the Th1 phenotype. Of the antigens tested, TpN37 appears to contribute the most to the Th1 cytokine response and therefore may play a key role in the clearance of T. pallidum from lesions. PMID- 10456929 TI - Phage-displayed T-cell epitope grafted into immunoglobulin heavy-chain complementarity-determining regions: an effective vaccine design tested in murine cysticercosis. AB - A new type of immunogenic molecule was engineered by replacing all three complementarity-determining-region (CDR) loops of the human immunoglobulin (Ig) heavy-chain variable (V(H)) domain with the Taenia crassiceps epitope PT1 (PPPVDYLYQT) and by displaying this construct on the surfaces of M13 bacteriophage. When BALB/c mice were immunized with such phage particles (PIgphage), a strong protection against challenge infection in very susceptible female hosts was obtained. When specifically stimulated, the in vivo-primed CD4(+) and CD8(+) T cells isolated from mice immunized with PT1, both as a free peptide and as the PIgphage construct, proliferated in vitro, indicating efficient epitope presentation by both major histocompatibility complex class II and class I molecules in the specifically antigen-pulsed macrophages used as antigen-presenting cells. These data demonstrate the immunogenic potential of recombinant phage particles displaying CDR epitope-grafted Ig V(H) domains and establish an alternative approach to the design of an effective subunit vaccine for prevention of cysticercosis. The key advantage of this type of immunogen is that no adjuvant is required for its application. The proposed strategy for immunogen construction is potentially suitable for use in any host-pathogen interaction. PMID- 10456930 TI - Identification of abundantly expressed novel and conserved genes from the infective larval stage of Toxocara canis by an expressed sequence tag strategy. AB - Larvae of Toxocara canis, a nematode parasite of dogs, infect humans, causing visceral and ocular larva migrans. In noncanid hosts, larvae neither grow nor differentiate but endure in a state of arrested development. Reasoning that parasite protein production is orientated to immune evasion, we undertook a random sequencing project from a larval cDNA library to characterize the most highly expressed transcripts. In all, 266 clones were sequenced, most from both 3' and 5' ends, and similarity searches against GenBank protein and dbEST nucleotide databases were conducted. Cluster analyses showed that 128 distinct gene products had been found, all but 3 of which represented newly identified genes. Ninety-five genes were represented by a single clone, but seven transcripts were present at high frequencies, each composing >2% of all clones sequenced. These high-abundance transcripts include a mucin and a C-type lectin, which are both major excretory-secretory antigens released by parasites. Four highly expressed novel gene transcripts, termed ant (abundant novel transcript) genes, were found. Together, these four genes comprised 18% of all cDNA clones isolated, but no similar sequences occur in the Caenorhabditis elegans genome. While the coding regions of the four genes are dissimilar, their 3' untranslated tracts have significant homology in nucleotide sequence. The discovery of these abundant, parasite-specific genes of newly identified lectins and mucins, as well as a range of conserved and novel proteins, provides defined candidates for future analysis of the molecular basis of immune evasion by T. canis. PMID- 10456932 TI - Induction of a rat enteric defensin gene by hemorrhagic shock. AB - Multicellular organisms utilize a battery of extracellular and cellular mechanisms to defend against microbial infiltration. Among the armamentarium used by the small intestine to defend against microbial invasion are antimicrobial peptides called defensins. We previously have shown that gut barrier function is impaired following hemorrhagic shock, resulting in translocation of bacteria or endotoxin. Using a rat model, we examined the effect of hemorrhagic shock on alpha-defensin expression. We utilized the anchored reverse transcriptase PCR strategy to isolate a rat enteric defensin cDNA. The cDNA is 406 bases in length and encodes a putative prepro-enteric defensin that we have named rat defensin 5 (RD-5). RD-5 expression is restricted to the small intestine and is specifically localized by in situ hybridization to the Paneth cells. A 10-fold increase in its steady state levels was observed in the distal intestine immediately after the termination of shock. This is the first study to show that enteric defensins are inducible following injury. We suggest that enteric defensins may contribute to the complex and integrated barrier function of the intestinal mucosal surface. PMID- 10456931 TI - Immunogenicity of DNA vaccines expressing tuberculosis proteins fused to tissue plasminogen activator signal sequences. AB - Novel tuberculosis DNA vaccines encoding native ESAT-6, MPT-64, KatG, or HBHA mycobacterial proteins or the same proteins fused to tissue plasminogen activator (TPA) signal sequences were evaluated for their capacity to elicit humoral, cell mediated, and protective immune responses in vaccinated mice. While all eight plasmids induced specific humoral responses, the constructs expressing the TPA fusions generally evoked higher antibody responses in vaccinated hosts. Although most of the DNA vaccines tested induced a substantial gamma interferon response in the spleen, the antigen-specific lung responses were 2- to 10-fold lower than the splenic responses at the time of challenge. DNA vaccines encoding the ESAT-6, MPT-64, and KatG antigens fused to TPA signal sequences evoked significant protective responses in mice aerogenically challenged with low doses of Mycobacterium tuberculosis Erdman 17 to 21 days after the final immunization. However, the protective response induced by live Mycobacterium bovis BCG vaccine was greater than the response induced by any of the DNA vaccines tested. These results suggest that the tuberculosis DNA vaccines were able to elicit substantial immune responses in suitably vaccinated mice, but further refinements to the constructs or the use of alternative immunization strategies will be needed to improve the efficacy of these vaccine candidates. PMID- 10456933 TI - Porin from Pseudomonas aeruginosa induces apoptosis in an epithelial cell line derived from rat seminal vesicles. AB - Micromolar concentrations of porin, purified from the outer membranes of Pseudomonas aeruginosa, induced in vitro the classic morphological and biochemical signs of apoptosis in an epithelial cell line (SVC1) derived from the rat seminal vesicle secretory epithelium. The programmed cell death (PCD) was p53 independent and associated with significant decrease of bcl-2 expression, a marked increase of c-myc transcriptional activity, and an absence of the mRNA coding for tissue transglutaminase. The Ca(2+) influx, caused by the porin treatment of SVC1 cells, appears to play an important role in the triggering of apoptosis in our biological model. The possibility that the porin property of inducing PCD plays a role in the infertility of individuals chronically infected by gram-negative bacteria is discussed. PMID- 10456934 TI - LcrV of Yersinia pestis enters infected eukaryotic cells by a virulence plasmid independent mechanism. AB - Yersinia pestis is the causative agent of bubonic plague and possesses a set of plasmid-encoded, secretable virulence proteins termed LcrV and Yops which are essential for survival in mammalian hosts. Yops and LcrV are secreted by a type III mechanism (Ysc), and Yops are unidirectionally targeted into the cytosol of associated eukaryotic cells in a tissue culture infection model. LcrV is required for Yops targeting, and recent findings have revealed that it can localize to the bacterial surface; however, its fate in this infection model has not been investigated in detail. In this study, we compared the localization of LcrV to that of the targeted proteins YopE and YopM by immunoblot analysis of fractions of Yersinia-infected HeLa cultures or by laser-scanning confocal microscopy of infected monolayers. Both LcrV and YopE were secreted by contact-activated, extracellularly localized yersiniae and were targeted to the HeLa cell cytosol. Although a significant amount of LcrV partitioned to the culture medium (unlike YopE), this extracellular pool of LcrV was not the source of the LcrV that entered HeLa cells. Unlike targeting of YopE and YopM, targeting of LcrV occurred in the absence of a functional Ysc apparatus and other virulence plasmid (pCD1) expressed proteins. However, the Ysc is necessary for LcrV to be released into the medium, and our recent work has shown that localization of LcrV on the bacterial surface requires the Ysc. These results indicate that two mechanisms exist for the secretion of LcrV by Y. pestis, both of which are activated by contact with eukaryotic cells. LcrV secreted by the Ysc reaches the bacterial surface and the surrounding medium, whereas the second is a novel, Ysc independent pathway which results in localization of LcrV in the cytosol of infected cells but not the surrounding medium. PMID- 10456935 TI - Antiphosphorylcholine antibody levels are elevated in humans with periodontal diseases. AB - Human immunoglobulin G2 (IgG2) serum concentrations and the IgG2 antibody response to Actinobacillus actinomycetemcomitans can be influenced by genes, by environmental factors such as smoking, and by periodontal disease status. Examination of the IgG2 response to phosphorylcholine (PC), a response thought to be mainly induced by the C polysaccharide of Streptococcus pneumoniae, suggested that periodontal disease status was also associated with this response. This prompted the hypothesis that PC is an important oral antigen associated with organisms in the periodontal flora and that anti-PC antibody is elevated as a consequence of periodontal disease. Subjects in various periodontal disease diagnostic categories in which attachment loss is exhibited were tested for anti PC in serum. Those with adult periodontitis, localized juvenile periodontitis, generalized early-onset periodontitis, and gingival recession all had similar levels of anti-PC IgG2 serum antibody which were significantly greater than in the group of subjects with no attachment loss. Analysis of plaque samples from subgingival and supragingival sites in all diseases categories for reactivity with the anti-PC specific monoclonal antibody TEPC-15 revealed that a substantial proportion of the bacteria in dental plaque (30 to 40%) bear PC antigen; this antigen was not restricted to morphotypes resembling only cocci but was also present on rods and branched filamentous organisms. We found that S. mitis, S. oralis, and S. sanguis, as well as oral actinomycetes, including A. viscosus, A. odontolyticus, and A. israelii, incorporated substantial amounts of [(3)H]choline from culture media. Further analysis of antigens derived from these organisms by Western blot indicated that S. oralis, S. sanguis, A. viscosus, A. odontolyticus, and A. israelii contained TEPC-15-reactive antigens. The data show that many commonly occurring bacterial species found in dental plaque contain PC antigen and that immunization with plaque-derived PC antigens as a consequence of inflammation and periodontal attachment loss may influence systemic anti-PC antibody concentrations. PMID- 10456936 TI - beta-Chemokines enhance parasite uptake and promote nitric oxide-dependent microbiostatic activity in murine inflammatory macrophages infected with Trypanosoma cruzi. AB - In the present study, we describe the ability of Trypanosoma cruzi trypomastigotes to stimulate the synthesis of beta-chemokines by macrophages. In vivo infection with T. cruzi led to MIP-1alpha, RANTES, and JE/MCP1 mRNA expression by cells from peritoneal inflammatory exudate. In addition, in vitro infection with T. cruzi resulted in expression of beta-chemokine MIP-1alpha, MIP 1beta, RANTES, and JE mRNA by macrophages. The expression of the beta-chemokine MIP-1alpha, MIP-1beta, RANTES, and JE proteins by murine macrophages cultured with trypomastigote forms of T. cruzi was confirmed by immunocytochemistry. Interestingly, macrophage infection with T. cruzi also resulted in NO production, which we found to be mediated mainly by beta-chemokines. Hence, treatment with anti-beta-chemokine-specific neutralizing antibodies partially inhibited NO release by macrophages incubated with T. cruzi parasites. Further, the addition of the exogenous beta-chemokines MIP-1alpha, MIP-1beta, RANTES, and JE/MCP-1 induced an increased T. cruzi uptake, leading to enhanced NO production and control of parasite replication in a dose-dependent manner. L-NMMA, a specific inhibitor of the L-arginine-NO pathway, caused a decrease in NO production and parasite killing when added to cultures of macrophages stimulated with beta chemokines. Among the beta-chemokines tested, JE was more potent in inhibiting parasite growth, although it was much less efficient than gamma interferon (IFN gamma). Nevertheless, JE potentiates parasite killing by macrophages incubated with low doses of IFN-gamma. Together, these results suggest that in addition to their chemotactic activity, murine beta-chemokines may also contribute to enhancing parasite uptake and promoting control of parasite replication in macrophages and may play a role in resistance to T. cruzi infection. PMID- 10456937 TI - Molecular cloning and characterization of rat genes encoding homologues of human beta-defensins. AB - beta-Defensins are cationic peptides with broad-spectrum antimicrobial activity that may play a role in mucosal defenses of several organs. They have been isolated in several species, and in humans, two beta-defensins have been identified. Here, we report the identification of two genes encoding beta defensin homologues in the rat. Partial cDNAs were found by searching the expressed-sequence-tag database, and primers were designed to generate full length mRNA coding sequences. One gene was highly similar to the human beta defensin-1 (HBD-1) gene and mouse beta-defensin-1 gene at both the nucleic acid and amino acid levels and was termed rat beta-defensin-1 (RBD-1). The other gene, named RBD-2, was homologous to the HBD-2 and bovine tracheal antimicrobial peptide (TAP) genes. The predicted prepropeptides were strongly cationic, were 69 and 63 residues in length for RBD-1 and RBD-2, respectively, and contained the six-cysteine motif characteristic of beta-defensins. The beta-defensin genes mapped closely on rat chromosome 16 and were closely linked to the alpha defensins genes, suggesting that they are part of a gene cluster, similar to the organization reported for humans. Northern blot analysis showed that both RBD-1 and RBD-2 mRNA transcripts were approximately 0.5 kb in length; RBD-1 mRNA was abundantly transcribed in the rat kidney, while RBD-2 was prevalent in the lung. Reverse transcription-PCR indicated that RBD-1 and RBD-2 mRNAs were distributed in a variety of other tissues. In the lung, RBD-1 mRNA expression localized to the tracheal epithelium while RBD-2 was expressed in alveolar type II cells. In conclusion, we characterized two novel beta-defensin homologues in the rat. The rat may be a useful model to investigate the function and contribution of beta defensins to host defense in the lung, kidney, and other tissues. PMID- 10456938 TI - The EspD protein of enterohemorrhagic Escherichia coli is required for the formation of bacterial surface appendages and is incorporated in the cytoplasmic membranes of target cells. AB - The formation of EspA-containing surface appendages in pathogenic Escherichia coli strains, both enteropathogenic E. coli (EPEC) and Shiga toxin-producing E. coli strains, is essential for critical events in the infective process, e.g., localized bacterial adherence to host cells with formation of microcolonies and induction of attaching and effacing lesions. It has been reported that EPEC mutants deficient in the production of EspD, which is encoded by the esp operon, are unable to accumulate actin underneath adherent bacteria but exhibit an attachment similar to that of the wild type. Here, we report the construction and characterization of an in-frame espD deletion mutant of the enterohemorrhagic E. coli (EHEC) strain EDL933. In contrast to what was observed in EPEC mutants, the EDL933 espD mutant not only lacked the capacity to accumulate actin but also exhibited an impaired attachment to HeLa cells. The synthesis of the EspD protein was also essential for the formation of EspA-containing filaments. Finally, localization studies demonstrated that the EspD protein is transferred to the cytoplasm and integrated into the cytoplasmic membranes of infected cells. These results help to elucidate the underlying molecular events in infections caused by EHEC. PMID- 10456939 TI - Human beta-defensin-1 mRNA is transcribed in tympanic membrane and adjacent auditory canal epithelium. AB - The external auditory canal is less susceptible to infections than the sensitive middle-ear cavity. Since recent research has provided insight to the production of potent antimicrobial peptides from various surface epithelia, we wanted to investigate whether protection of the external auditory canal in part could be explained by the production of human beta-defensin-1 (HBD-1). This particular peptide is known to be constitutively expressed in various surface epithelia, such as airway, skin, and urogenital tissues. By reverse transcriptase PCR we demonstrate HBD-1 mRNA in the pars tensa and pars flaccida of the tympanic membrane and in the meatal skin. In situ hybridization studies localized the HBD 1 mRNA to the epidermal layer of these tissues. The HBD-1 transcripts were also evident in the sebaceous glands and in hair follicles of the meatal skin. In contrast, HBD-1 mRNA was not detected in the tympanal epithelium of the eardrum. The widespread presence of mRNA encoding for this broad-spectrum antimicrobial peptide in the meatal skin and tympanic membrane suggests that HBD-1 participates in the innate antimicrobial defense of the external auditory canal and middle-ear cavity. PMID- 10456940 TI - Cell surface-exposed tetanus toxin fragment C produced by recombinant Bacillus anthracis protects against tetanus toxin. AB - Bacillus anthracis, the causal agent of anthrax, synthesizes two surface layer (S layer) proteins, EA1 and Sap, which account for 5 to 10% of total protein and are expressed in vivo. A recombinant B. anthracis strain was constructed by integrating into the chromosome a translational fusion harboring the DNA fragments encoding the cell wall-targeting domain of the S-layer protein EA1 and tetanus toxin fragment C (ToxC). This construct was expressed under the control of the promoter of the S-layer component gene. The hybrid protein was stably expressed on the cell surface of the bacterium. Mice were immunized with bacilli of the corresponding strain, and the hybrid protein elicited a humoral response to ToxC. This immune response was sufficient to protect mice against tetanus toxin challenge. Thus, the strategy developed in this study may make it possible to generate multivalent live veterinary vaccines, using the S-layer protein genes as a cell surface display system. PMID- 10456941 TI - The 102-kilobase pgm locus of Yersinia pestis: sequence analysis and comparison of selected regions among different Yersinia pestis and Yersinia pseudotuberculosis strains. AB - We report the complete 119,443-bp sequence of the pgm locus from Yersinia pestis and its flanking regions. Sequence analysis confirms that the 102-kb unstable pgm locus is composed of two distinct parts: the pigmentation segment and a high pathogenicity island (HPI) which carries virulence genes involved in iron acquisition (yersiniabactin biosynthetic gene cluster). Within the HPI, three genes coding for proteins related to phage proteins were uncovered. They are located at both extremities indicating that the entire HPI was acquired en bloc by phage-mediated horizontal transfer. We identified, within the pigmentation segment, two novel loci that may be involved in virulence: a fimbriae gene cluster and a locus probably encoding a two component regulatory system similar to the BvgAS regulatory system of Bordetella pertussis. Three genes containing frameshift mutations and two genes interrupted by insertion element insertion were found within this region. To investigate diversity among different Y. pestis and Yersinia pseudotuberculosis strains, the sequence of selected regions of the pgm locus and flanking regions were compared from 20 different Y. pestis and 10 Y. pseudotuberculosis strains. The results showed that the genes interrupted in Y. pestis are intact in Y. pseudotuberculosis. However, one of these mutations, in the bvgS homologue, is only present in Y. pestis strains of biovar Orientalis and not in those of the biovars Antiqua and Medievalis. The results obtained by analysis of variable positions in the sequence are in accordance with historical records, confirming that biovar Orientalis is the most recent lineage. Furthermore, sequence comparisons among 29 Yersinia strains suggest that Y. pestis is a recently emerged pathogen that is probably entering the initial phase of reductive evolution. PMID- 10456942 TI - B- and T-cell immune responses to pneumococcal conjugate vaccines: divergence between carrier- and polysaccharide-specific immunogenicity. AB - Conjugation of various serotypes of pneumococcal polysaccharide (PnPS) to carrier protein enhances the magnitude of the polysaccharide-specific antibody response, presumably by eliciting T-cell help. However, variability in PnPS serotype specific immunogenicity has been observed. CBA/J mice immunized with either 6B or 19F PnPS conjugated to the protein carrier Cross Reactive Material(197) (CRM(197)) produce a strong anti-PnPS antibody response; however, when mice are immunized with 23F PnPS conjugated to CRM(197), they fail to produce a significant anti-PnPS response. In order to determine whether this difference was related to alterations in antigen processing of the carrier protein and the subsequent T-cell responses, we studied proliferation of lymphocytes from CBA/J mice immunized with CRM(197) alone or conjugated to 6B, 19F, or 23F PnPS. T-cell proliferative responses to synthetic peptides demonstrated that lymph node cells elicited by the poorly immunogenic conjugate 23F-CRM(197) recognized many, but not all, of the epitopes recognized by lymph node cells elicited by 6B- and 19F CRM(197) as well as additional epitopes. Despite marked differences in PnPS specific immunogenicity, all mice made high titers of CRM(197) antibodies of the immunoglobulin G(1) isotype. Cells from mice immunized with any of the conjugates yielded vigorous T-cell responses to whole antigen. We conclude that the serotype of PnPS can alter the peptide specificities of T-cell responses, but even a poorly immunogenic PnPS conjugate can elicit a significant T-cell response. Thus, conjugation of PnPS to a carrier protein that elicits carrier-specific T- and B cell responses does not necessarily enhance PnPS immunogenicity. PMID- 10456943 TI - Control of filament formation in Candida albicans by polyamine levels. AB - Candida albicans, the most common fungal pathogen, regulates its cellular morphology in response to environmental conditions. The ODC gene, which encodes ornithine decarboxylase, a key enzyme in polyamine biosynthesis, was isolated and disrupted. Homozygous null Candida mutants behaved as polyamine auxotrophs and grew exclusively in the yeast form at low polyamine levels (0.01 mM putrescine) under all conditions tested. An increase in the polyamine concentration (10 mM putrescine) restored the capacity to switch from the yeast to the filamentous form. The strain with a deletion mutation also showed increased sensitivity to salts and calcofluor white. This Candida odc/odc mutant was virulent in a mouse model. The results suggest a model in which polyamine levels exert a pleiotrophic effect on transcriptional activity. PMID- 10456944 TI - Contribution of Salmonella typhimurium virulence factors to diarrheal disease in calves. AB - Limited knowledge is available about the virulence mechanisms responsible for diarrheal disease caused by Salmonella typhimurium. To assess the contribution to diarrheal disease of virulence determinants identified in models of infection, we tested a collection of S. typhimurium mutants for their ability to cause enteritis in calves. S. typhimurium strains carrying mutations in the virulence plasmid (spvR), Salmonella pathogenicity island 2 (SPI-2) (spiB), or SPI-5 (sopB) caused mortality and acute diarrhea in calves. An S. typhimurium rfaJ mutant, which is defective for lipopolysaccharide outer core biosynthesis, was of intermediate virulence. Mutations in SPI-1 (hilA and prgH) or aroA markedly reduced virulence and the severity of diarrhea. Furthermore, histopathological examination of calves infected with SPI-1 or aroA mutants revealed a marked reduction or absence of intestinal lesions. These data suggest that virulence factors, such as SPI-1, which are required during intestinal colonization are more important for pathogenicity in calves than are genes required during the systemic phase of S. typhimurium infection, including SPI-2 or the spv operon. This is in contrast to the degree of attenuation caused by these mutations in the mouse. PMID- 10456945 TI - Activation of caspase 3 during Legionella pneumophila-induced apoptosis. AB - The hallmark of Legionnaires' disease is replication of Legionella pneumophila within cells in the alveolar spaces. The mechanisms by which L. pneumophila replicates intracellularly and kills the host cell are largely not understood. We have recently shown that within 3 h of initiation of the infection and prior to intracellular replication, L. pneumophila induces apoptosis in macrophages, alveolar epithelial cells, and peripheral blood monocytes, which correlates with cytopathogenicity (L.-Y. Gao and Y. Abu Kwaik, Infect. Immun. 67:862-870, 1999). In this report, we show that the ability of L. pneumophila to induce apoptosis is, largely, not growth phase regulated. We demonstrate that the induction of apoptosis by L. pneumophila in macrophages is mediated through the activation of caspase 3. The enzymatic activity of caspase 3 to cleave a specific synthetic substrate in vitro is detected in L. pneumophila-infected macrophages at 2 h after infection and is maximal at 3 h, with over 900% increase in activity. The activity of caspase 3 to cleave a specific substrate [poly(ADP-ribose) polymerase, or PARP] in vivo is also detected at 2 h and is maximal at 3 h postinfection. The activity of caspase 3 to cleave the synthetic substrate in vitro and PARP in vivo is blocked by a specific inhibitor of caspase 3. The kinetics of caspase 3 activation correlates with that of L. pneumophila-induced nuclear apoptosis. Inhibition of caspase 3 activity blocks L. pneumophila-induced nuclear apoptosis and cytopathogenicity during early stages of the infection. Consistent with the ability to induce apoptosis, extracellular L. pneumophila also activates caspase 3. Three dotA/icmWXYZ mutants of L. pneumophila that are defective in inducing apoptosis do not induce caspase 3 activation, suggesting that expression and/or export of the apoptosis-inducing factor(s) is regulated by the dot/icm virulence system. This is the first description of the role of caspase 3 activation in induction of nuclear apoptosis in the host cell infected by a bacterial pathogen. PMID- 10456946 TI - Observed differences in virulence-associated phenotypes between a human clinical isolate and a veterinary isolate of Mycobacterium avium. AB - Mycobacterium avium, the most common opportunistic pathogen in patients with AIDS, is frequently isolated from a variety of environmental sources, but rarely can these environmental isolates be epidemiologically linked with isolates known to cause human disease. Using a number of in vitro tissue culture assays, we found significant pathogenic differences between a serotype 4 human clinical M. avium isolate and a serotype 2 veterinary isolate. Cell association of the patient strain with a human intestinal cell line was 1.7 times that of the veterinary strain. Growth of this clinical strain in human peripheral blood mononuclear cell-derived macrophages increased from 12-fold higher than that of the veterinary isolate after 2 days to 200-fold higher after 4 days. By the conclusion of each experiment, lysis of all examined host cell types and accumulation of cell debris were observed in infections with the human isolate, but monolayers remained relatively intact in the presence of the animal isolate. The two strains also differed in the ability to stimulate human immunodeficiency virus replication in coinfected host cells, with p24 antigen levels after 6 days threefold higher in the cells coinfected with the clinical strain than in those infected with the veterinary strain. If the genetic differences responsible for the phenotypes observed in these assays can be identified and characterized, it may be possible to determine which M. avium strains in the environment are potential human pathogens. PMID- 10456947 TI - Use of genetically manipulated strains of Clostridium perfringens reveals that both alpha-toxin and theta-toxin are required for vascular leukostasis to occur in experimental gas gangrene. AB - A hallmark of gas gangrene (clostridial myonecrosis) pathology is a paucity of leukocytes infiltrating the necrotic tissue. The cause of this paucity most likely relates to the observation of leukocyte aggregates at the border of the area of tissue necrosis, often within the microvasculature itself. Infecting mice with genetically manipulated strains of Clostridium perfringens type A (deficient in either alpha-toxin or theta-toxin production) resulted in significantly reduced leukocyte aggregation when alpha-toxin was absent and complete abrogation of leukocyte aggregation when theta-toxin was absent. Thus, both alpha-toxin and theta-toxin are necessary for the characteristic vascular leukostasis observed in clostridial myonecrosis. PMID- 10456948 TI - Adhesion of Escherichia coli to HeLa cells mediated by Trypanosoma cruzi surface glycoprotein-derived peptides inserted in the outer membrane protein LamB. AB - Peptides derived from the surface glycoprotein gp82 of Trypanosoma cruzi, previously implicated in the parasite's invasion of host cells, were expressed as fusions to the protein LamB of Escherichia coli in a region known to be exposed on the cell surface. Bacteria expressing these proteins adhered to HeLa cells in a manner that mimics the pattern of parasite invasion of mammalian cells. Purified LamB fusion proteins were shown to bind to HeLa cells and to inhibit infection by T. cruzi, supporting the notion that these gp82-derived peptides can mediate interaction of the parasite with its host. PMID- 10456949 TI - Role of complement receptors in uptake of Mycobacterium avium by macrophages in vivo: evidence from studies using CD18-deficient mice. AB - Mycobacterium avium is an intracellular pathogen that has been shown to invade macrophages by using complement receptors in vitro, but mycobacteria released from one cell can enter a second macrophage by using receptors different from complement receptors. Infection of CD18 (beta(2) integrin) knockout mice and the C57 BL/6 control mice led to comparable levels of tissue infection at 1 day, 2 days, 1 week, and 3 weeks following administration of bacteria. A histopathological study revealed similar granulomatous lesions in the two mouse strains, with comparable numbers of organisms. In addition, transmission electron microscopy of spleen tissues from both strains of mice showed bacteria inside macrophages. Our in vivo findings support the hypothesis that M. avium in the host is likely to use receptors other than CR3 and CR4 receptors to enter macrophages with increased efficiency. PMID- 10456950 TI - Inhibitory effects of protamines on proteolytic and adhesive activities of Porphyromonas gingivalis. AB - Protamines (salmine prepared from sperm DNA of salmon and clupeine from herring sperm), which are basic peptides rich in arginine, were found to inhibit the proteolytic activity of arginine-specific cysteine protease (RC-protease) from Porphyromonas gingivalis. Lineweaver-Burk plot analysis revealed that the protamines competitively inhibited proteolytic activity with cleavage of benzoyl L-arginine-p-nitroanilide, a synthetic substrate of RC-protease. Furthermore, the protamines were capable of binding strongly to P. gingivalis fimbriae and inhibited fimbrial interaction with immobilized fibronectin. These results clearly show that protamines are potent inhibitors of the proteolytic and adhesive activities of P. gingivalis. PMID- 10456951 TI - Role of local cytokines in increased gastric expression of the secretory component in Helicobacter pylori infection. AB - Using immunohistochemical staining, we examined the presence of secretory component (SC) on epithelial cells in gastric and duodenal biopsy specimens collected from Helicobacter pylori-infected individuals and healthy controls. Gastric epithelial cells from healthy volunteers expressed low, but detectable, levels of SC. In contrast, significantly higher level of expression of SC (P < 0.001) was observed on epithelial cells in the antra of H. pylori-infected individuals. The antral SC expression correlated with staining for gamma interferon of intraepithelial and lamina propria lymphocytes (r(s) = 0.76 and 0.69, respectively, P < 0.001) and correlated weakly with production of tumor necrosis factor alpha (r(s) = 0.43, P < 0.05), but it did not correlate at all with interleukin-4 production. PMID- 10456953 TI - Virulence of Mycobacterium tuberculosis CDC1551 and H37Rv in rabbits evaluated by Lurie's pulmonary tubercle count method. AB - The virulence of the CDC1551 strain of Mycobacterium tuberculosis was compared to that of H37Rv in a rabbit inhalation model. While rabbits that inhaled the two strains produced equal numbers of grossly visible primary tubercles, CDC1551 tubercles were smaller and contained fewer bacilli than H37Rv tubercles. These findings suggest that a miniepidemic near the Kentucky-Tennessee border caused by CDC1551 was due not to increased virulence but to increased transmissibility. PMID- 10456952 TI - Molecular mechanism for the spontaneous generation of pigmentless Porphyromonas gingivalis mutants. AB - Porphyromonas gingivalis is one of the pathogens associated with periodontal diseases, and its protease activity has been implicated as an important virulence factor. Kgp is the major Lys-gingipain protease of P. gingivalis and appears to be involved not only in enzyme activity but also in hemagglutination and the pigmented phenotype due to heme accumulation and/or hemoglobin binding. However, little information concerning the molecular mechanism for the spontaneous generation of pigmentless P. gingivalis mutants is currently available. In this study, several spontaneous pigmentless mutants of P. gingivalis were isolated and characterized. The results revealed that a portion of the kgp gene had been deleted from the chromosomes of the pigmentless mutants. This deletion appears to result from recombination between the highly homologous DNA sequences encoding the adhesin domains of the tandemly arranged hagA and kgp genes on the chromosomes of P. gingivalis strains. PMID- 10456954 TI - Age-associated differences in immunoglobulin G1 (IgG1) and IgG2 subclass antibodies to pneumococcal polysaccharides following vaccination. AB - Immunoglobulin G (IgG) subclass antibody responses to pneumococcal vaccines were determined for human subjects in four age groups. The ratios of IgG1/IgG2 antibody concentrations declined with advancing age for all five of the serotypes tested. Protein-conjugate vaccines elicited enhanced IgG antibody responses over plain polysaccharide vaccines in infants but not in adult groups. PMID- 10456955 TI - Coinfection with Toxoplasma gondii inhibits antigen-specific Th2 immune responses, tissue inflammation, and parasitism in BALB/c mice infected with Leishmania major. AB - Lesion size, cellular infiltration, and tissue parasitism in the footpads of BALB/c mice infected with Leishmania major were all dramatically inhibited during acute but not chronic infection with Toxoplasma gondii. Similarly, acute but not chronic toxoplasmosis at the time of infection with L. major had a strong inhibitory effect on development of acquired immune responses mediated by Th2 lymphocytes. In contrast, no major changes in Leishmania-specific Th1-mediated responses were observed in mice coinfected with T. gondii. PMID- 10456956 TI - Identification of a third metalloprotease toxin gene in extraintestinal isolates of Bacteroides fragilis. AB - To further understand the epidemiology of enterotoxigenic Bacteroides fragilis (ETBF), 89 extraintestinal B. fragilis strains from Seoul, Korea, were examined for secretion of B. fragilis toxin (BFT) by the HT29/C1 biologic assay and for the B. fragilis toxin gene (bft) by colony blot hybridization and PCR. Complete agreement between the three techniques was found. Overall, 34 B. fragilis strains (38%) were identified as ETBF. Eleven of the 34 ETBF strains (32%) expressed a new isoform of BFT (Korea-BFT). This new isoform is more related to BFT-2 than to BFT-1. Like BFT-1 and BFT-2, Korea-BFT cleaves E-cadherin, the zonula adherens protein. PMID- 10456957 TI - Differential regulation of enteric and systemic salmonellosis by slyA. AB - Mutation of slyA, which reduces Salmonella typhimurium virulence in mice, caused only minor attenuation of S. typhimurium virulence in orally inoculated calves. This correlated with modest reductions in intestinal invasion and enteropathogenic responses in bovine ligated ileal loops. slyA appears to regulate virulence genes involved in systemic, but not enteric, salmonellosis. PMID- 10456958 TI - Bactericidal and cross-protective activities of a monoclonal antibody directed against Neisseria meningitidis NspA outer membrane protein. AB - The cross-bactericidal and cross-protective activities of a monoclonal antibody (MAb) named Me-7, which is directed against an antigenically highly conserved epitope on the meningococcal NspA protein, were studied. This MAb efficiently killed in vitro, in the presence of rabbit or human serum, 13 of 14 meningococcal strains tested, including 9 of 9, 2 of 3, and 2 of 2 strains of serotypes B, A, and C, respectively. MAb Me-7 also significantly reduced by more than 75% the levels of bacteremia recorded for mice challenged with 10 of 11 meningococcal strains tested. Analysis of the predicted amino acid sequence of the NspA protein from the meningococcal strain MCH88 (A:4:P1.10), which was not killed by MAb Me 7, indicated the presence of an additional glutamine residue at position 73, compared to the three other NspA sequences. The data presented in this study suggest that antibodies directed against this highly conserved outer membrane protein could protect against meningococcal infections. PMID- 10456959 TI - Outer membrane proteins Omp10, Omp16, and Omp19 of Brucella spp. are lipoproteins. AB - The deduced sequences of the Omp10, Omp16, and Omp19 outer membrane proteins of Brucella spp. contain a potential bacterial lipoprotein processing sequence. After extraction with Triton X-114, these three proteins partitioned into the detergent phase. Processing of the three proteins is inhibited by globomycin, a specific inhibitor of lipoprotein signal peptidase. The three proteins were radioimmunoprecipitated from [(3)H]palmitic acid-labeled Brucella abortus lysates with monoclonal antibodies. These results demonstrate that Omp10, Omp16, and Omp19 are lipoproteins. PMID- 10456960 TI - Immune cells are required for cutaneous ulceration in a swine model of chancroid. AB - Cutaneous lesions of the human sexually transmitted genital ulcer disease chancroid are characterized by the presence of intraepidermal pustules, keratinocyte cytopathology, and epidermal and dermal erosion. These lesions are replete with neutrophils, macrophages, and CD4(+) T cells and contain very low numbers of cells of Haemophilus ducreyi, the bacterial agent of chancroid. We examined lesion formation by H. ducreyi in a pig model by using cyclophosphamide (CPA)-induced immune cell deficiency to distinguish between host and bacterial contributions to chancroid ulcer formation. Histologic presentation of H. ducreyi induced lesions in CPA-treated pigs differed from ulcers that developed in immune competent animals in that pustules did not form and surface epithelia remained intact. However, these lesions had significant suprabasal keratinocyte cytotoxicity. These results demonstrate that the host immune response was required for chancroid ulceration, while bacterial products were at least partially responsible for the keratinocyte cytopathology associated with chancroid lesions in the pig. The low numbers of H. ducreyi present in lesions in humans and immune-competent pigs have prevented localization of these organisms within skin. However, H. ducreyi organisms were readily visualized in lesion biopsies from infected CPA-treated pigs by immunoelectron microscopy. These bacteria were extracellular and associated with necrotic host cells in the epidermis and dermis. The relative abundance of H. ducreyi in inoculated CPA treated pig skin suggests control of bacterial replication by host immune cells during natural human infection. PMID- 10456961 TI - Molecular cloning of the Pasteurella haemolytica pomA gene and identification of bovine antibodies against PomA surface domains. AB - The gene (pomA) encoding PomA, an OmpA-like major outer membrane protein of the bovine respiratory pathogen Pasteurella haemolytica, was cloned, and its nucleotide sequence was determined. The deduced amino acid sequence of PomA has significant identity with the sequences of other OmpA family proteins. Absorption of three different bovine immune sera with whole P. haemolytica cells resulted in a reduction of bovine immunoglobulin G reactivity with recombinant PomA in Western immunoblots, suggesting the presence of antibodies against PomA surface domains. PMID- 10456962 TI - Experimental transmission of Neisseria gonorrhoeae from pregnant rat to fetus. AB - Sprague-Dawley rats were infected on day 20 of pregnancy by intraperitoneal inoculation with Neisseria gonorrhoeae. Disseminated gonococcal infection (DGI) and pelvic inflammatory disease (PID) strains in the presence of C1q but not in the presence of bovine serum albumin (BSA) were able to spread from the pregnant rat to the fetus and resulted in fetal mortality. Transmission of DGI and PID strains that are serum resistant (ser(r)) and sac-4 positive but not of a local infection strain that is ser(s) and sac-4 negative was facilitated by the C1q dependent mechanism. This study provides the first experimental model that may mimic the transmission of gonococcal infection from mother to the fetus during pregnancy. PMID- 10456963 TI - Blood mononuclear cell nitric oxide production and plasma cytokine levels in healthy gabonese children with prior mild or severe malaria. AB - Plasmodium falciparum malaria is an important cause of morbidity and mortality in children. Factors that determine the development of mild versus severe malaria are not fully understood. Since host-derived nitric oxide (NO) has antiplasmodial properties, we measured NO production and NO synthase (NOS) activity in peripheral blood mononuclear cells (PBMC) from healthy Gabonese children with a history of prior mild malaria (PMM) or prior severe malaria (PSM) caused by P. falciparum. The PMM group had significantly higher levels of NOS activity in freshly isolated PBMC and higher NO production and NOS activity in cultured PBMC. The investigation of NO-modulating cytokines (e.g., interleukin 12, gamma interferon, tumor necrosis factor alpha [TNF-alpha], and transforming growth factor beta1) as an explanation for differing levels of NOS activity revealed that plasma levels of TNF-alpha were significantly higher in the PSM group. Our results suggest that NOS/ NO and TNF-alpha are markers for prior disease severity and important determinants of resistance to malaria. PMID- 10456964 TI - Step 3 of the asthma guidelines. PMID- 10456965 TI - Pulmonary thromboendarterectomy for chronic thromboembolic pulmonary hypertension. PMID- 10456966 TI - Extended tumour necrosis factor/HLA-DR haplotypes and asthma in an Australian population sample. AB - BACKGROUND: Tumour necrosis factor (TNF) is a potent pro-inflammatory cytokine which is prominent in asthmatic airways. TNF shows genetic variations in secretion which are linked to polymorphisms in the TNF gene complex and the surrounding major histocompatibility (MHC) locus. These polymorphisms do not seem to be themselves functionally important. In these circumstances, the identification of disease associated haplotypes (combination of alleles on individual chromosomes) may narrow the search for polymorphisms which alter gene function. METHODS: TNF-308, LTalpha NcoI, and HLA-DRB1 polymorphisms were investigated for association with asthma, bronchial responsiveness, and medication use in 1004 subjects in 230 families from a general population sample. RESULTS: The common LTalpha NcoI*1/TNF-308*2/HLA-DRB1*03 haplotype, which was present in 11% of unrelated individuals, was weakly associated with asthma (OR = 1.38, p = 0.016, corrected for familial correlation). The rarer LTalpha NcoI*1/TNF-308*2/HLA-DRB1*02 haplotype, which was found in 0.6% of unrelated subjects, was more strongly associated with asthma (OR = 6.68, p = 0.002). This haplotype also showed association with bronchial hyperresponsiveness (OR = 21.9, p = 0. 0000) and the use of inhaled or oral steroids (OR 8.0, p = 0.04). CONCLUSIONS: The results of this study show only two extended TNF/HLA-DR haplotypes to be associated with asthma. The search for functional alleles responsible for an increased risk of asthma should concentrate on the LTalpha NcoI*1/TNF-308*2/HLA-DRB1*02 haplotype. PMID- 10456967 TI - Administration of the BCG vaccination using the multipuncture method in schoolchildren: a comparison with the intradermal method. AB - BACKGROUND: BCG vaccination using the multipuncture device (the Heaf gun) is recommended in the UK for infants and very small children only. The aim of this study was to investigate the rate of conversion of the tuberculin test, the safety and acceptability of BCG vaccination using the multipuncture device and to compare it with the conventional intradermal method in schoolchildren. METHODS: Schoolchildren attending schools in Tower Hamlets who were eligible for BCG vaccination were tuberculin tested using the Heaf gun. Those with grade 0-1 reaction were randomised to receive BCG vaccination using either the multipuncture or the intradermal method. The site of BCG vaccination was inspected after eight weeks for inflammatory changes and scarring. A questionnaire about pain and inflammation at the site of vaccination was completed. The Heaf test was repeated at eight weeks and its results were assessed by an examiner unaware of the results of the previous Heaf test and the method of BCG administration. The Heaf test conversion was deemed to have occurred if there was a change of at least one grade in the response. RESULTS: One hundred and sixty nine children (83 girls) of mean age 11.8 years completed the study, of which 81 received BCG by the multipuncture method. The Heaf test did not convert in 22 of 81 (27. 2%) receiving BCG by the multipuncture device compared with six of 88 (6.8%) who received the vaccine by the intradermal method (odds ratio 0.2, 95% confidence interval 0.07 to 0.55). The BCG scar was visible in all children who had intradermal BCG compared with 67 of 81 (81.8%) of the multipuncture group. The multipuncture method was less painful and caused fewer inflammatory changes than the intradermal method. CONCLUSIONS: In schoolchildren the multi-puncture device for administering BCG caused a lower rate of tuberculin conversion as measured by the Heaf test and less of an inflammatory response than the intradermal method. The method needs to be modified before it is applied on a wider scale to schoolchildren. PMID- 10456968 TI - Effect of brachial plexus co-activation on phrenic nerve conduction time. AB - BACKGROUND: Diaphragm function can be assessed by electromyography of the diaphragm during electrical phrenic nerve stimulation (ES). Whether phrenic nerve conduction time (PNCT) and diaphragm electrical activity can be reliably measured from chest wall electrodes with ES is uncertain. METHODS: The diaphragm compound muscle action potential (CMAP) was recorded using an oesophageal electrode and lower chest wall electrodes during ES in six normal subjects. Two patients with bilateral diaphragm paralysis were also studied. Stimulations were deliberately given in a manner designed to avoid or incur co-activation of the brachial plexus. RESULTS: For the oesophageal electrode the PNCT was similar with both stimulation techniques with mean (SE) values of 7.1 (0.2) and 6.8 (0.2) ms, respectively (pooled left and right values). However, for surface electrodes the PNCT was substantially shorter when the brachial plexus was activated (4.4 (0.1) ms) than when it was not (7.4 (0.2) ms) (mean difference 3.0 ms, 95% CI 2.7 to 3.4, p<0.0001). A small short latency CMAP was recorded from the lower chest wall electrodes during stimulation of the brachial plexus alone. CONCLUSIONS: The results of this study show that lower chest wall electrodes only accurately measure PNCT when care is taken to avoid stimulating the brachial plexus. A false positive CMAP response to phrenic stimulation could be caused by inadvertent stimulation of the brachial plexus. This finding may further explain why the diaphragm CMAP recorded from chest wall electrodes can be unreliable with cervical magnetic stimulation during which brachial plexus activation occurs. PMID- 10456969 TI - Inflammatory cells and cellular activation in the lower respiratory tract in Churg-Strauss syndrome. AB - BACKGROUND: To obtain insight into the mechanisms of tissue injury in lung disease due to Churg-Strauss syndrome (CSS), the bronchoalveolar lavage (BAL) cell profile and the levels in the BAL fluid of cell products released by activated eosinophils and neutrophils were assessed. METHODS: Thirteen patients with active progressive CSS (n = 7) or CSS in partial remission (n = 6) underwent clinical staging and bronchoalveolar lavage. The levels of eosinophil cationic protein (ECP), myeloperoxidase (MPO), and peroxidase activity in the BAL fluid were determined and the results were compared with those of 19 patients with pulmonary active Wegener's granulomatosis (WG) and nine control subjects. RESULTS: In patients with progressive CSS the BAL cell profile was dominated by eosinophils, neutrophil elevation being the exception. The eosinophilia was associated with high ECP levels (4.39 ng/ml and 0. 40 ng/ml in the two CSS groups compared with unmeasurable values in the controls). Individual patients with highly active CSS also had raised MPO levels, comparable to the levels in the most active WG patients. Peroxidase activity in the BAL fluid was 1.26 U/ml and 0. 10 U/ml in the two groups of patients with CSS and 0.20 U/ml in the controls. Pulmonary disease in patients with WG was characterised by an extensive increase in MPO (0.30 ng/ml versus 0.13 ng/ml in the controls) together with high peroxidase activity in the BAL fluid (4. 37 U/ml), but only a small increase in ECP levels was seen. No correlation was found between the ECP and MPO levels in patients with CSS which suggests that eosinophil and neutrophil activation vary independently of each other. CONCLUSIONS: These findings suggest that, in addition to eosinophil activation, neutrophil activation is an important feature in some patients with highly active CSS. The balance of neutrophil and eosinophil involvement appears to be variable and this may be one explanation for the individually variable treatment requirements of patients with CSS. PMID- 10456971 TI - The open circuit nitrogen washout technique for measuring the lung volume in infants: methodological aspects. AB - BACKGROUND: Lung volume measurement by nitrogen washout is widely used in infants, though a lack of accuracy and changes of calibration over time have been reported. The potential sources of error were explored in order to increase the accuracy and reliability of the technique. METHODS: A commercial system for nitrogen washout and a 0.5 litre calibrating syringe as a lung model were used to perform over 2000 in vitro washouts, including simulated rapid breathing, shallow breathing, periodic breathing, sighs, and brief apnoeas. A constant 10 l/min bias flow of oxygen and extended equipment warming times were employed. A collapsible breathing bag was incorporated into the washout circuit. Following a single two point calibration, known air volumes from 42 ml to 492 ml were measured by nitrogen washout over a 14 hour period. The flow waveform in the nitrogen mixing chamber during a washout in vitro, with and without the breathing bag in the circuit, was also studied. RESULTS: The mean coefficient of variation of all volumes was 0.66%. The mean difference between measured and known volumes was 0.30 ml (95% confidence interval (CI) -0.18 to 0.79). This difference was not statistically significant (p = 0.22). The mean percentage error was -0.1% (range 0.47% to 0.46%). Nitrogen calibration remained stable for 14 hours. Without the breathing bag flow transients were frequent in the mixing chamber during in vitro washout. CONCLUSIONS: This technique increases the accuracy in vitro and the precision in vivo of volume measurement by nitrogen washout. Sources of potential errors including baseline drifting and inadequate equipment warming times were identified. The breathing bag acted as a buffer reservoir, preventing large swings in flows within the nitrogen mixing chamber during washouts, and should be an integral component of the nitrogen washout circuit. PMID- 10456970 TI - Lung volume reduction surgery (LVRS) for chronic obstructive pulmonary disease (COPD) with underlying severe emphysema. AB - BACKGROUND: Lung volume reduction surgery (LVRS) has recently re-emerged as a surgical option for the treatment of end stage chronic obstructive pulmonary disease (COPD) due to underlying severe emphysema. Advocates of LVRS claim that it represents a significant breakthrough in the management of this challenging group of patients while sceptics point to uncertainty about the effectiveness of the operation. METHODS: A systematic review was conducted of the evidence on the effects of LVRS in patients with end stage COPD secondary to severe emphysema. RESULTS: The most rigorous evidence on the effectiveness of LVRS came from case series. Seventy five potentially relevant studies were identified and 19 individual series met the methodological criteria for inclusion. The pattern of results was consistent across individual studies despite a significant degree of clinical heterogeneity. Significant short term benefits occurred across a range of outcomes which appeared to continue into the longer term. Physiological improvements were matched by functional and subjective improvements. Early mortality rates were low and late mortality rates compared favourably with those of the general COPD population. However, the entire research base for the intervention is subject to the limitations of study designs without parallel control groups. CONCLUSIONS: LVRS appears to represent a promising option in the management of patients with severe end stage emphysema. However, until the results of ongoing clinical trials are available, the considerable uncertainty that exists around the effectiveness and cost effectiveness of the procedure will remain. PMID- 10456972 TI - Role of NO in recovery from neonatal hypoxic pulmonary hypertension. AB - BACKGROUND: The management of sick newborn infants who have sustained a hypoxic insult is a common clinical problem but relatively little is known about the recovery process. The aim of this study was to investigate this process in newborn piglets. METHODS: Thirty five newborn piglets were exposed to chronic hypobaric hypoxia for three days, either from birth, three or 14 days of age, and were allowed to recover for one, three, or six days. Control animals of relevant age were also studied. The heart weight ratio and pulmonary arterial muscularity were measured. Endothelial dependent and independent relaxation of the isolated intrapulmonary conduit arteries was determined in classical organ chamber studies, together with measurement of basal and stimulated cGMP accumulation. RESULTS: After six days of recovery the hypoxia induced right ventricular hypertrophy and pulmonary arterial medial hypertrophy had decreased in all animals but values were still abnormal in the two younger age groups. Relaxation was still impaired during the first three days of recovery in all groups, had normalised by six days in the two youngest groups, but relaxation (both endothelium dependent and independent) remained impaired in older animals. In these older animals basal nitric oxide (NO) production and basal and stimulated cGMP accumulation was normal. CONCLUSIONS: The recovery of the smooth muscle cells lags behind that of the endothelial cells. A normal stimulated increase in cGMP with reduced relaxation suggests an altered threshold for cGMP effected relaxation. These findings help to explain why some hypoxic infants require protracted NO therapy. PMID- 10456973 TI - Reduction of bleomycin induced lung fibrosis by transforming growth factor beta soluble receptor in hamsters. AB - BACKGROUND: Transforming growth factor beta (TGF-beta) is a key mediator of collagen synthesis in the development of lung fibrosis. It has previously been shown that the administration of TGF-beta antibody and TGF-beta binding proteoglycan, decorin, reduced bleomycin (BL) induced lung fibrosis in animals. The present study was carried out to investigate whether intratracheal instillation of TGF-beta soluble receptor (TR) would minimise the BL induced lung fibrosis in hamsters. METHODS: The effect of a recombinant TR (TGFbetaRII) on the lung collagen accumulation was evaluated in a BL hamster model of pulmonary fibrosis. Animals were divided into four groups and intratracheally injected with saline or BL at 6.5 U/4 ml/kg followed by intratracheal instillation of phosphate buffered saline (PBS) or 4 nmol TR in 0.3 ml twice a week. Twenty days after the first intratracheal instillation the hamsters were killed for bronchoalveolar lavage (BAL) fluid, biochemical, and histopathological analyses. RESULTS: Treatment of hamsters with TR after intratracheal instillation of BL significantly reduced BL induced lung fibrosis as shown by decreases in the lung hydroxyproline level and prolyl hydroxylase activity, although they were still significantly higher than those of the saline control. Histopathological examination showed a considerable decrease in BL induced fibrotic lesions by TR treatment. However, TR did not prevent the BL induced increases in total cells and protein in the BAL fluid. CONCLUSIONS: These results suggest that TR has antifibrotic potential in vivo and may be useful in the treatment of fibrotic diseases where increased TGF-beta is associated with excess collagen accumulation. PMID- 10456974 TI - Issues at the interface between primary and secondary care in the management of common respiratory disease. 3: Providing better asthma care: what is there left to do? PMID- 10456975 TI - Genetics and pulmonary medicine.10: Genetic epidemiology of pulmonary function. PMID- 10456977 TI - Pulmonary hypertension secondary to neurofibromatosis: intimal fibrosis versus thromboembolism. AB - Neurofibromatosis has been known to involve blood vessels throughout the body. Pulmonary involvement with interstitial fibrosing alveolitis has been described but no case of pulmonary vascular involvement has been reported to date. A 51 year old patient with cutaneous neurofibromatosis is described who presented with severe pulmonary hypertension and radiographic, scintigraphic, and angiographic evidence of chronic thromboembolic pulmonary hypertension. Severe intimal fibrosis consistent with vascular involvement with neurofibromatosis was found on endarterectomy with no evidence of pulmonary thromboembolism. Neurofibromatosis of pulmonary arteries should be considered as a possible cause of pulmonary hypertension. PMID- 10456978 TI - Laryngeal aspergillosis following high dose inhaled fluticasone therapy for asthma. AB - The case history is presented of a 75 year old man with chronic asthma who was treated with inhaled fluticasone propionate in a daily dose of 2 mg using a Diskhaler. After three years of treatment he developed progressive hoarseness. Both vocal cords were colonised by Aspergillus fumigatus which formed a white slough on the surface. Biopsy specimens showed changes suggestive of laryngeal aspergillosis with an ulcerated epithelium, fibrinopurulent debris, and colonies of fungal hyphae. A slow recovery occurred after three months of treatment with topical amphotericin and with cessation of inhaled corticosteroids. Laryngoscopy is recommended if hoarseness occurs during treatment with fluticasone. PMID- 10456979 TI - Implanted octacalcium phosphate (OCP) stimulates osteogenesis by osteoblastic cells and/or committed osteoprogenitors in rat calvarial periosteum. AB - Our previous studies demonstrated that the octacalcium phosphate (OCP) causes new appositional bone formation on the OCP when implanted into the subperiosteal region of murine calvaria. The OCP may stimulate the cell population committed to the osteoblastic differentiation in the periosteum and have them express the phenotype. The present study was designed to investigate which periosteal cell population is involved in bone formation on the OCP with applying the OCP implants on top of and underneath the periosteum. The periosteum of the rat parietal bones was flapped and the OCP was implanted on top of or underneath the periosteum, in which the implantation sites were defined using the membrane filter. The histology was examined to see if new appositional bone formation occurs on the OCP implant under each condition. New bone was deposited on the OCP on the bone surface separated from the periosteum by the filter, whereas no bone was formed either under the periosteum separated from the bone surface by the filter or on the periosteum. The present study suggests that the OCP acts on osteoblasts, bone lining cells and/or their closely committed progenitors on the bone surface to express the phenotype and deposit new bone on the OCP implant. PMID- 10456980 TI - The influence of maternal hypothyroidism and radioactive iodine on rat embryonal development: thyroid C-cells. AB - There have been no works devoted to the study of the influence of (131)I and maternal (131)I-induced hypothyroidism on the state of the C-cells in the thyroid gland of the developing embryos. A study was made on the effect of a dose of 150 microCi (131)I (0.5 Gy) leading to hypothyroidism in rats, on 35 mother rats and 168 newborn pups. The mother rats were divided into control and four treated groups which were injected with (131)I before pregnancy, on gestation days 5, 10, and 16, respectively. Immunohistochemically, the thyroid gland was examined for calcitonin-positive cells. Maternal hypothyroidism induced by (131)I leads to the development of hyperplasia and hyperthrophy of calcitonin-positive cells in the pups at the time of birth. The discovery of separate C-cells in the peripheral zone of the thyroid lobe may be evidence of an unbalance in the development of the medial and lateral source of the thyroid. There is a verifiable increase in the quantity of C-cells per 1 mm(2) field of the localization in the central zone of the gestation days 10 and 16 groups. This might be a compensatory mechanism for regulating the activity of the thyroid gland under induced hypothyroidism. Thus, in cases when there is a breakdown in the normal external regulation of the embryonic morphogenesis, a reduction in the level of maternal thyroid hormones and also direct exposure to (131)I, there is also a change in the foetus' internal regulatory systems. A change in C-cell system could lead to the appearance of endocrinological disorders later in life. PMID- 10456981 TI - Bone turnover associated with antler growth in red deer (Cervus elaphus). AB - Although it is known that skeletal bone depletion occurs during antler growth in deer, it is not clear whether repletion of the skeleton takes place before or after completion of antler development. This study attempted to correlate repeated scanning electron microscopic measures of ilium and rib bone porosity from six approximately 2-monthly biopsy samples (using back-scattered imaging) and biochemical markers of bone turnover (serum hydroxyproline and osteocalcin concentrations) taken for 11 months with antler growth in six red deer stags. No changes were detected in ilium samples but changes in porosity of rib bones and an elevation of the biochemical markers indicated that skeletal depletion occurred during the antler growth period. However, the decrease in rib bone porosity and decline in markers of bone turnover took place before completion of antler growth, indicating that a considerable amount of skeletal repletion could have occurred whilst antlers were also undergoing bone accretion. This latter finding extends the current view of antler growth being accompanied by a form of reversible osteoporosis in the skeleton by showing that there is a period when the antlers and skeleton are both undergoing net bone formation. PMID- 10456982 TI - Morphological changes of the myenteric plexus during early postnatal development of the rat. AB - The enteric nervous system needs to adapt itself constantly to the postnatal changes of the developing gut. The aim of this study was to examine the morphological changes between the distal and proximal segments of the gastrointestinal (GI) tract during the first two postnatal weeks. Myenteric plexus from the duodenum, proximal and distal colon of 1-, 7- and 14-day-old rat pups was dissected and examined under the scanning electron microscope. Wholemounts from the same regions and postnatal stages were stained with cuprolinic blue. Neuronal numbers per ganglionic area were counted and neuronal sizes were measured. Furthermore, segments of the above-mentioned areas were embedded in resin and semithin sections were cut. The thickness of the circular and longitudinal muscle layers was measured. The morphology of the myenteric plexus depends on localization as well as on the age of the animal. While in younger animals the myenteric plexus is usually densely packed, the network expands with increasing age. Similarly, the thickness of the circular and the longitudinal muscle layers increases. Nerve cell numbers per ganglionic area increase from duodenum to distal colon and decrease from the 1-day (P1) to the 14 day-old (P14) animal. The longest diameters and the area of the nerve cells decrease from duodenum to distal colon and increase with age of the animal. The intensity of the cuprolinic blue staining varies also according to age and segment of the gut. During the first two postnatal weeks the three-dimensional architecture of the myenteric plexus as well as the size and densities of the enteric neurons change according to the increasing gut length and the thickness of the muscle layer. The differences between duodenum and colon might reflect the different physiological properties of the proximal and distal gut as well as a varying grade of maturity, which is also supported by a variation in the cuprolinic blue staining intensity. PMID- 10456983 TI - Anatomical characteristics and three-dimensional model of the dog dorsal lateral geniculate body. AB - The morphological and laminar characteristics of the dorsal lateral geniculate nucleus (LGN) and medial interlaminar nucleus (MIN) of the domestic dog (Canis familiaris) were studied by three-dimensional computer reconstruction of labeled retinal afferents following intraocular HRP injections. As previously reported, the dog LGN consisted of layers A, A1, C, C1, C2, and C3. Layers A, C, and C2 receive contralateral-eye inputs, and layers A1 and C1 ipsilateral inputs. The dog MIN was found to have four orderly interdigitating layers; layers 1, 2, 3, and 4, medial to lateral. MIN layers 1 and 3 received contralateral inputs, and layers 2 and 4 ipsilateral inputs. Layer 1 had the largest soma of all LGN/MIN layers. LGN layer A was partially separated into medial and lateral subdivisions by a cleft free of somata. The overall three-dimensional shape of the lateral geniculate body was like the letter C, with the convex part of the C directed posteriorly. The relative volume of the MIN was smaller than in the cat; the canine MIN comprised 8.3% of the combined volume of layers A, A1 and the MIN, while that of the cat comprised 14.2% as estimated from Sanderson's map. The volume of all contralateral-eye layers, combining both LGN and MIN, was 31.2 mm(3) (78%), and that for ipsilateral layers was 8.6 mm(3) (22%). The ratio of ipsilateral to contralateral laminar volumes is much lower in the dog than in the cat. PMID- 10456984 TI - Immunolocalization and expression of prohibitin, a mitochondrial associated protein within the rat ovaries. AB - This study was designed to determine the cellular distribution and pattern of expression for the mitochondria-associated protein, prohibitin, during the transitional stages of follicular differentiation within the rat ovary. Immunohistochemical staining techniques were used on frozen sections to examine the localization of prohibitin to preantral, antral, preovulatory, and atretic follicles. Prohibitin localization was also determined in corpus luteum from adult rats, in addition to those from infant and juvenile ovaries, before and after gonadotropin stimulation. Western and Northern blotting techniques were used for qualitative and quantitative assessment of prohibitin expression levels within the ovary. Prohibitin was localized within granulosa cells of infant and juvenile ovaries in a relatively heterogeneous staining pattern. The oocyte also exhibited robust prohibitin expression at all stages of follicular development. In addition, strong prohibitin expression was evident in the corpus luteum as well as in follicles undergoing atresia. Additional data derived from studies involving a GnRH-agonist indicate that increases in prohibitin protein expression correlate with the initial events of apoptosis. Collectively, these results support a growth regulatory role for prohibitin within the rat ovary. Therefore, we propose that prohibitin may serve as an important regulator of granulosa cell fate during follicular development. PMID- 10456985 TI - The AV junction region of the heart: a comprehensive study correlating gross anatomy and direct three-dimensional analysis. Part I. Architecture and topography. AB - There is little detailed knowledge of the architecture of the AV junction region, the cytoarchitecture of the AV node or of its atrial connections. In the present study, the gross anatomy and topography of intracardiac structures in 21 adult canine hearts were photographically compared in whole and dissected hearts and tissue blocks and serial histologic sections made in three orthogonal planes. There are seven major new findings: 1) A coronary sinus fossa exists at the crux of the heart. It separates the right medial atrial wall (MAW) superoposterior region from the left atrium, its floor is the coronary sinus, and it carries the medial atrionodal bundle and proximal AV bundle on its right wall. 2) The posterior MAW forms two isolated bridges of myocardium as it surrounds the coronary sinus ostium, is isolated from the sinus venarum with crista terminalis and interatrial septum-by the floor of the inferior vena cava, and the narrow bridges link the posterior atrial wall to the mid MAW. 3) The tendon of Todaro has both epicardial and endocardial exposures, terminates in the superoposterior MAW and its medial aspect is adjacent sequentially to the medial atrionodal bundle and proximal AV bundle. 4) Only ordinary myocardium contacts the anulus fibrosus. 5) The ventricular septum's shoulder is humped shape posteriorly, is completely overlaid by anular myocardium and the medial leaflet and is joined by struts of papillary muscle. 6) The membranous septum joins the anterior ventricular septum to the crista supraventricularis, forms part of the posterior noncoronary and right aortic valve sinus walls and encases the right bundle branch. 7) The specialized conduction tissues, the superior, medial and lateral atrionodal bundles, the proximal AV bundle, AV node, distal AV bundle and right bundle branch are subjacent to MAW epicardium outside the right atrium, share regular intracardiac relationships with topographic landmarks and the medial atrionodal bundle, terminal superior atrionodal bundle, the proximal AV bundle and AV node are aligned to the medial leg of Koch's triangle. Thus, atrial myocardium of the AV junction region is that of the MAW. The floor of the inferior vena cava forms a natural barrier to impulse transmission along the full extent of the posterior MAW. The specialized tissues are outside of the MAW. Anatomic landmarks form reliable topographic landmarks for the specialized AV junction region tissues. A knowledge of the association of the specialized conduction tissues with specific regions of the MAW is useful in localizing the tissues and along with the coronary sinus fossa provides several extracardiac approaches. PMID- 10456986 TI - Selective and transient expression of a native chondroitin sulfate epitope in Deiters' cells, pillar cells, and the developing tectorial membrane. AB - The tectorial membrane (TM) is an acellular connective tissue overlying the sensory hair cells of the organ of Corti. Association of the tectorial membrane with the stereocilia of the sensory hair cells is necessary for proper auditory function. During development, the mature tectorial membrane is thought to arise by fusion of a "major" and "minor" tectorial membrane (Lim, Hear Res 1986;22:117 146). Several proteins and glycoconjugates have been detected in the developing TM; however, the specific molecules which mediate fusion of the two components of the TM have not been identified. In the present study, a novel monoclonal antibody (TC2) that recognizes a native epitope on glycosaminoglycans enriched in chondroitin-4-sulfate revealed a transient and restricted expression in the developing gerbil TM. The localization patterns suggest that Deiters' and pillar cells secrete a TC2-positive matrix prior to birth that later becomes incorporated into the marginal band and superior layer (cover net) of the TM. The developmental timecourse and patterns of TC2 reactivity suggest that this molecule may play a critical role in the fusion of the minor TM with the major TM. PMID- 10456987 TI - Enzyme histochemical localization of Na(+),K(+)-ATPase and NADH-DE in the developing rat parotid gland. AB - Information on ductal differentiation in the developing rat parotid gland is sparse. Striated and excretory ducts are rich in a number of enzymes related to ion movement. The objective of this investigation was to delineate histochemically the chronology of two of these, ouabain-sensitive Na(+),K(+) ATPase and NADH-DE, in the developing rat parotid gland. Parotid glands were excised from rats at representative ages from 20 days in utero to 42 days. Enzyme histochemistry was performed on air-dried frozen sections. For Na(+), K(+) ATPase, some sections also were fixed in phosphate-buffered formalin. Ouabain blocked Na(+),K(+)-ATPase activity, and neither enzyme reacted without substrate. Weak Na(+),K(+)-ATPase reactions were initially seen in unfixed sections at 1 day, and increased steadily to the adult pattern of strong (concentrated basolaterally) in striated ducts and excretory ducts, respectively, and weak to modest (diffuse) in acini and intercalated ducts at 28 days. In fixed sections, localization was sharper but the reaction was somewhat reduced. NADH-DE was modest in terminal buds and ducts before birth, then progressively changed to the adult pattern of weak in acini and intercalated ducts and strong (concentrated basally and luminally) in striated and excretory ducts at 28 days. As demonstrated by enzyme histochemistry of Na(+),K(+)-ATPase and NADH-DE, differentiation of rat parotid striated ducts and excretory ducts occurs mainly between birth and 28 days. Anat Rec 256:72-77, 1999. Published 1999 Wiley-Liss, Inc. PMID- 10456988 TI - Number of neurons in the retinal ganglion cell layer of the quokka wallaby do not change throughout life. AB - During adult life, the topography of the retinal pigment epithelium (RPE) of the quokka wallaby changes gradually. Cells in peripheral retina enlarge in surface area while those in mid-temporal retina, adjacent to the area centralis, a high density region in the ganglion cell layer, decrease in area, implying that the tissue in this area is drawing together. We speculated that high ganglion cell densities in temporal regions might be maintained, in the face of cell loss due to aging, by this apparent drawing together of the RPE sheet. Therefore, we examined the retinal ganglion cell layer of the quokka in cresyl violet stained wholemounts from animals aged from 0. 55 to 13.5 years. We found that total neuron number in the retinal ganglion cell layer of the quokka did not decrease significantly throughout life even though individuals in captivity live long lives (9-15 years). Ganglion and amacrine cells were counted separately and identified by strict morphological criteria. Nevertheless, the proportion of ganglion to amacrine cells appeared to decrease linearly throughout life, indicating that the morphology of a proportion of neurons became more amacrine like during aging. Mean cell size did not change throughout life. In the quokka, retinal area increases slowly throughout life and may account for the small reduction in cell density seen in most retinal regions. PMID- 10456989 TI - Evidence of a phenotypically determined ductal cell lineage in mouse salivary glands. AB - The submandibular salivary gland of mice contains a parenchymal element, the granular duct, which matures peripubertally from the striated ducts. Granular duct cells also differentiate from intercalated ducts in the adult mouse submandibular gland. Using preproNGF-A as a signature protein of mature granular duct cells, this study inquired if phenotypic determination might have occurred earlier than the first signs of cellular differentiation. Results from RT-PCR indicate the presence of preproNGF-A transcripts at all postnatal stages of development of the submandibular glands, as well as in adult sublingual glands. The preproNGF-A transcript was also detected prenatally as early as embryonic day 17 in the submandibular/sublingual complex. Using an antibody directed specifically against the "pre" peptide, immunocytochemistry showed preproNGF-A localized in the granular ducts and striated ducts of the adult submandibular gland. In addition preproNGF-A was detected throughout the first order branches of the intercalated duct system. In the neonatal gland, preproNGF-A was found in the large tubules that differentiate to the striated ducts. The early appearance of preproNGF-A in the histological lineage that sequentially gives rise to striated ducts and then to granular ducts suggests that this lineage is phenotypically determined as early as birth. An undifferentiated stage of the phenotypically determined lineage also appears to be retained in the intercalated duct system to provide progenitors for subsequent differentiation in the adult gland. Throughout development of the sublingual gland, preproNGF-A was detectable in the striated ducts or in their predecessors, suggesting that they may also represent a phenotypically determined cell lineage similar to that of the submandibular gland. PMID- 10456991 TI - Non satellite veins in the pelvic limb of the llama (Lama glama). AB - The objective of this study was to determine the major differences in the venous system of the pelvic limb of the llama (Lama glama) and that of other mammals, including humans. Eight adult llamas, preserved by means of 6% formalin solution at 0 degrees C, were dissected. The venous system was perfused with a solution of 17% coloured industrial gelatin. Two venous systems are recognised, superficial and deep. The veins of the deep system follow the same course as the arteries, except for two important veins located in the femoral, popliteal and crural regions. A more voluminous vein than the femoral originates in a venous tripod in the proximal part of the femoral canal and forms an anastomosis with the femoral vein distally. The popliteal vein gives off an expansion located along the popliteal region. In the leg, it continues with a vein that accompanies the tibial nerve and forms an anastomosis distally with the medial saphenous vein. The described venous distribution represents an interesting complement to the medial saphenous, popliteal and femoral veins, being more important than the last one for its volume. In the thigh it would correspond to the deep femoral vein as in humans and dogs; the homology in the popliteal and leg regions is too difficult because it probably corresponds to the caudal tibial vein of the human, the caudal branch of the medial saphenous vein, and the satellite vein of the tibial nerve in the horse. PMID- 10456990 TI - Fluorescent double-label study of lateral reticular nucleus projections to the spinal cord and periaqueductal gray in the rat. AB - Following injections of WGA-HRP into either the spinal cord or periaqueductal gray, labeled neurons were observed bilaterally along the periphery of the lateral reticular nucleus (LRN) magnocellular division. The possibility that some of these neurons in the LRN provide collateral axonal branches to both the periaqueductal gray and the spinal cord was investigated in rats using a retrograde double-labeling method employing two different fluorescent tracers, True Blue and Nuclear Yellow. Following sequential injection of the two fluorescent axonal tracers into the spinal cord and periaqueductal gray in the same animal, a modest number of double-labeled neurons were observed bilaterally near the medial and dorsal perimeter of the magnocellular division of the LRN. The labeled neurons were distinctly multipolar in shape and measured approximately 15-18 mu in their greatest transverse diameter. No double-labeled neurons were observed in the parvocellular or subtrigeminal divisions of the LRN. Based upon these observations, it is suggested that collaterals of the LRN-spinal pathway provide feedback information to the periaqueductal gray that might then be used to modulate the participation of the latter cell group in a variety of pain processing and cardiovascular regulatory functions. PMID- 10456992 TI - Blood and marrow transplantation activity in Europe 1997. European Group for Blood and Marrow Transplantation (EBMT). AB - Data from the annual survey on transplant activity 1997, collected from 457 transplant teams in 31 European countries by the European Group for Blood and Marrow Transplantation (EBMT) were used to describe current status and to assess relative and absolute changes in indication, donor type and stem cell source compared to 1991. A total of 16950 patients were reported to have a first blood or marrow transplant in 1997, a total of 18 923 procedures, including re- and double transplants were performed. Of the 16950 first transplants, 4751 (28%) were allogeneic, 12199 (72%) autologous transplants. Of the autologous transplants, 829 (7%) were bone marrow derived, 11370 (93%) from peripheral blood stem cells or combined bone marrow and peripheral blood stem cell transplants. Of the allogeneic transplants, 3311 (70%) were bone marrow, 1440 (30%) were peripheral blood stem cell transplants. In 1991, the respective figures were 2175 allogeneic (44%) and 2786 (56%) autologous transplants, more than 90% of the autologous, all allogeneic transplants bone marrow derived. Main indications in 1997 were leukemias with 5253 transplants (31%), 70% allogeneic; lymphomas with 6773 transplants (40%), 94% autologous; solid tumors with 4154 transplants (24%), 99% autologous; non-malignant disorders with 770 transplants (5 %), 85 % allogeneic. There was an absolute increase of 11971 transplants since 1991. An increase was observed in all disease categories. Marked differences were found, when the relative increase index (RII) for specific disease categories over time was analyzed. In allogeneic transplants, relatively more transplants were performed in 1997 for acute myeloid leukemia beyond 1st complete remission (RII 1.28), myelodysplastic syndromes (RII 1.58), chronic lymphocytic leukemia (RII 1.33) and non-Hodgkin's lymphoma (RII 1.58). For autologous transplant indications, a high relative increase index was observed in myelodysplastic syndromes (RII 3.77), in multiple myeloma (RII 2.12) and carcinoma of the breast (RII 6.37) with a relative decrease in leukemias (RII 0.39) and certain solid tumors such as glioma (RII 0.27) and neuroblastoma (RII 0.46). These data present the current status of blood and marrow transplantation in Europe. They show the change from bone marrow to blood as stem cell source and highlight shifts in indication. They provide a basis for patient counselling and health care planning. PMID- 10456993 TI - [The European community, the euro and science]. PMID- 10456994 TI - [Training of local health managers: process of identification of strategic actions]. AB - In Brazil, in recent years, as a result of the increasing participation of county authorities in health care, a need for tools which would contribute to the better preparation of local administrators, complementary to the activities of a more academic nature, has been recognized. One of the possible alternatives is the exploitation of experiences, regarded as successful, in local health care planning and administration, by using them as material for "case studies" in activities with selected groups of health care administrators thus, stimulating the identification of those elements which contributed to the favorable results attained, and their interactions, in the quest for analogies which would facilitate the identification of new perspectives for their own situations. In this article an experience of a "case studies" development, in response to a demand from UNICEF, based on 8 counties in from the North and Northeast, which were successful in using a "focus approach" in their organization of data and their utilisation in a seminar with 21 local managers from both regions, is presented. During the seminar the local health managers attained greater knowledge of the strategies implemented and identified feasible intervention alternatives. The methodological proposition of teaching on the basis of case studies, using a conceptual strategy of grouping experiences according to specific dimensions enabled local health managers to learn from their practical experiences. PMID- 10456995 TI - Chagas disease in an area of recent occupation in Cochabamba, Bolivia. AB - INTRODUCTION: A descriptive, entomological and seroepidemiological study on Chagas disease was conducted in a place of recent occupation on the outskirts of Cochabamba, Bolivia: Avaroa/Primer de Mayo (population:3,000), where the socio economic level is low and no control measures have been made available. METHODS: The immunofluorescent antibody test (IFAT) was used for IgG and IgM anti Trypanosoma cruzi antibodies in filter paper bloodspot eluates from 128 subjects (73 females, 55 males) selected by systematic sampling. Concerning each subject age, gender, birthplace, occupation, duration of residence and building materials used in their houses were recorded. Vectors were captured both in domestic and peridomestic environments. RESULTS: Seropositive, 12.5% (16/128): females, 15.1% (11/73); males, 9.1% (5/55). Average time of residence: 6.1 years for the whole population sample and 7.4 years for the seropositive subjects. Most houses had adobe walls (76. 7%, n= 30), galvanized iron rooves (86.7%) and earthen floors (53. 4%) 80% of the walls had crevices. One hundred forty seven specimens of Triatoma infestans were captured, of which 104 (70.7%) were domestic, and 1 peridomestic Triatoma sordida. Precipitin host identification: birds, 67.5%; humans, 27.8%; rodents, 11.9%; dogs, 8. 7%; cats, 1.6%. House infestation and density indices were 53.3 and 493.0 respectively. We found 21 (14.3%) specimens of T. infestans infected with trypanosomes, 18 (85.7%) of which in domestic environments. DISCUSSION: The elements for the vector transmission of Chagas disease are present in Avaroa/Primer de Mayo and the ancient custom of keeping guinea pigs indoors adds to the risk of human infection. In neighboring Cochabamba, due to substandard quality control, contaminated blood transfusions are not infrequent, which further aggravates the spread of Chagas disease. Prompt action to check the transmission of this infection, involving additionally the congenital and transfusional modes of acquisition, is required. PMID- 10456996 TI - [Reproductive risk factors associated with breast cancer in Colombian women]. AB - INTRODUCTION: As of 1992, breast cancer has been the second cause of death in Columbian women, with a rising trend in mortality due to this type of neoplasm (average annual rate 4.5*/100.000 inhabitants). Information about potential risk factors for breast cancer in Latin American countries is scarce. The objectives of the project were to test the breastfeeding protection against breast cancer and to establish the reproductive factors associated with breast cancer in Columbia. METHODS: A hospital case-control study was carried out from July 1995 to March 1996 in Bogota, Columbia, using paring by age groups. The study population consisted of 171 histopathologically confirmed cases and 171 controls. Reproductive history and sociodemographic data were collected through a questionnaire, and logistic regression models were used for statistical analysis of the data. RESULTS: The following associated factors were found as principal results: nuli-parousness as compared with women with over 3 children (OR=3.35 CI 1.40-8.0), age at first birth (OR=1.83 CI 0.70- 4.80), breast cancer history, breastfeeding the first child (OR=0.09 CI 0.01-0.64 for 1-11 months) and with a highly significant trend for accumulated breastfeeding above 24 months (p=0.001). CONCLUSIONS: This study indicates the importance of focusing on the promotion of prolonged breastfeeding by women identified as being at higher risk, and confirms that socio-economic level can determine life styles and reproductive events among women (such as breastfeeding time); this could explain the increase in breast cancer mortality in Latin American countries, similar to that in developed countries in terms of fertility and risk factors for breast cancer. The epidemiological information produced by this study will be useful for planning and carrying out early diagnosis and treatment of breast cancer in women identified as being at high risk of this disease. PMID- 10456997 TI - [Prevalence of use of medicines during pregnancy and its relationship to maternal factors]. AB - INTRODUCTION: There is in Brazil a growing use of industrially produced medicines even during the female reprodutive cycle. During pregnancy two organisms are exposed to the effects of medicines but in the foetus this may result in toxicities with possible irreversible lesions. The present study aims at verifying the prevalence of the use of medicines during pregnancy in the studied population and its relationship with maternal characteristics, the pharmacological groups used and the source of prescription. METHODS: The use of medication was evaluated among 1,620 women that gave birth in five public, private and contracted hospitals in the city of S. Paulo between July and September 1993. RESULTS AND CONCLUSIONS: The relation between maternal schooling and kind of assistance in hospitals has revealed social inequality in the access to the several kinds of delivery assistance services. The prevalence of use of at least one medicine was 97.6% with an average of 4.2 drugs per woman. The prevalence of use of drugs by medical prescription and self-medication were 94.9% and 33.5%, respectively. The most used medications, excluding vitamins, mineral salts and vaccines, were analgesics, antacids, antispasmodics and antemetics. Users of the largest number of medicines were, characteristically, over 29, married, of university level, salaried workers and had access to private health services. Medical assistance had a facilitating role in access to the use of drugs during the gestational period of the population studied. Women need to be made aware of the potential risks they expose their foetuses to when using so many medicines. Physicians should reflect on their role in contributing to the solution to this problem. PMID- 10456976 TI - Cytokines in asthma. PMID- 10456998 TI - [Cancer mortality in urban-industrial area of the Baixada Santista, Sao Paulo, (Brazil)]. AB - INTRODUCTION: In order to study the relationship between cancer and industrialization the cancer mortality in the an urban area (Baixada Santista, Brazil), an important industrial-harbour complex, two distinct groups: industrialized and non-industrialized, is analyzed. METHODS: A total of 8,546 cancer death registers (according to ICD-9) in males over 10 years old, resident in the that area, were obtained from "Fundacao Sistema Estadual de Analise de Dados (SEADE)", during the period 1980-1993. The average mortality rates, standardized on the basis of the to 1960 world population for the area studied, and their strata: Stratum I (inside the industrial area - Santos, S. Vicente, Cubatao and Guaruja) and Stratum II (outside the industrial area - Praia Grande, Mongagua, Itanhaem and Peruibe) and the ratios among these rates were calculated adopting the 95% confidence interval. RESULTS: The annual average mortality rate was high, 197.9/100,000 and there was a statistically significant difference between the mortality rates observed in Stratum I and those in Stratum II, 209.2 and 146.7/100,000, respectively, with a ratio of 1.42 (CI 1.36 - 1.51). CONCLUSIONS: It seems that environmental and occupational exposure to carcinogenic chemical agents, related to the productive process in the industrial complex, is a significant factor in mortality due to cancer. PMID- 10456999 TI - [Mortality among Japanese migrants living in the State of Parana, Brazil]. AB - INTRODUCTION: Taking as a premise that the study concerning the morbimortality of migrant populations may make a contribution to a better understanding of the epidemiology of diseases, mainly the chronic-degenerative ones and their related risk factors, the aim of this study is an analysis of the mortality experience of a population of migrants born in Japan, but living in the State of Parana, Brazil, and a comparison of their pattern of mortality with those of Japan and Parana. METHODS: The population studied was composed of the Japanese migrants - Issei - living in the State of Parana, Brazil, of more than 50 years of age, identified in the X General Brazilian Census of 1st September, 1991. Information on deaths which occurred between 1st March 1990 and 28th February, 1993, was obtained through the Ministry of Health data base. The main causes of death were analysed after the calculation of the age-adjusted death rates, using the world standard population older than 50 years old, for each sex, for the Issei, the inhabitants in Japan and in Parana. Standardized Risk Ratio - SRR - and respective 95% confidence interval were estimated for selected causes among Issei versus the population living in Japan and Issei versus persons living in Parana. RESULTS: Among the main results, it was observed that the female Issei mortality rate was in an intermediary position when compared to Japan's and Parana's rates, while men showed figures quite close to the Japanese rates. Concerning the specific causes, it was observed that among male Issei, when compared to the Japanese population, the rates were significantly lower for stomach, but higher for diabetes and ischaemic heart diseases. Regarding the female Issei aged 50 or more years old, only the death rate for lung cancer was itself significantly lower than that of Japanese women. When compared to the Parana pattern, the male Issei rates were lower regarding ischaemic heart and cerebrovascular diseases. Concerning stomach and lung cancer, there was no statistical difference. The female Issei rates were lower for lung cancer and ischaemic heart diseases. Regarding diabetes and cerebrovascular diseases, no significant difference among rates was detected. CONCLUSIONS: The results obtained make it possible to assert a deviation in the Issei pattern of mortality from that of their country of origin (Japan) and a perseptible approximation to the pattern of their new homeland (Parana). Such observations suggest the influence of socio-cultural factors, mainly dietary habits, on their morbi-mortality. PMID- 10457000 TI - [Evaluation of the quality of mortality information by unspecified accidents and events of undetermined intent]. AB - INTRODUCTION: Mortality data are important for monitoring violence, making it possible to assess the trends and the impact of interventions towards its reduction. The objective of the study is to assess the quality of the filling out and codification of the death certificates for unspecified accidents and events of undetermined intent in the city of S. Paulo in 1996. METHODS: Death certificates on which the underlying cause of death (UCD) given was an unspecified accident (ICD-10 X59) or an event of undetermined intent (ICD-10 Y10 Y34) were selected for investigation at the Legal Medicine Institute (IML). After consulting the police reports which accompany the corpses to the IML, the autopsy reports and other legal forms, these were analysed and the UCD was recoded. RESULTS: For unspecified accidents, 53.2% were changed to a specified cause: 15.1% due to pedestrians injured in traffic accidents, 17.5% due to other traffic accidents and 14.5% due to falls. Homicides and suicides constituted 9.8%. In 20.9% no additional information was found. For events of undetermined intent, 2/3 had no clarification; in 1/3 of the cases, the underlying cause changed to falls (10.6%), homicides (7.5%) and pedestrians injured in transport accidents (6. 7%). CONCLUSIONS: The quality of mortality information by external causes in the City of S. Paulo is not satisfactory. The IML has not used all the available information to fill out the death certificates. The findings reveal that the instruction of the World Health Organization and the Brazilian Center for the Classification of Diseases to codify as accidents those events for which there is no information on the death certificate about the external cause, does not seem to be appropriate. In that category 66.0% of the deaths were found to have been inferred incorrectly as accidental. The improvement of the quality of mortality data due to external causes may contribute to the monitoring of violence and may give support to decisions leading to its reduction whatever the form that violence may take. PMID- 10457001 TI - [Problems in the standardization of the polymerase chain reaction for the diagnosis of pulmonary tuberculosis]. AB - INTRODUCTION: The recent increase in the number of tuberculosis cases has called the world's attention once again to a perennial health problem, especially prevalent in developing countries. The time elapsed between the diagnosis and the institution of therapy is an obstacle to tuberculosis control and there is an urgent need for the development of techniques for the disease's rapid diagnosis. To achieve this goal, molecular biology techniques have been exhaustively investigated. This work describes the use of a polymerase chain reaction for rapid diagnosis of tuberculosis in a developing country. The sensitivity and specificity of this technique is compared to standard techniques used in the microbiology laboratory. METHODS: This study was undertaken in Ribeirao Preto, S. Paulo State, Brazil. Forty-two sputum samples from suspected cases of tuberculosis attending the municipal health care centers were sent to the microbiology laboratory. The samples were processed for the detection of Mycobacterium tuberculosis by acid-fast bacilli determination, culture in Lowenstein-Jensen medium, and by a polymerase chain reaction that amplified a fragment of 123 base pairs of the Mycobacterium tuberculosis genome. RESULTS: Of the forty-two samples studied, one was contaminated and excluded from the study, ten were culture positive, ten were positive for the presence of acid-fast bacilli, and sixteen were polymerase chain reaction positive. The sensitivity and specificity of this technique were 90% and 81%, respectively. CONCLUSIONS: The polymerase chain reaction presented a sensitivity comparable to the culture and the whole procedure took only one day to complete. The results presented here make it a strong candidate for rapid diagnosis of tuberculosis in clinical settings making it possible to begin the specific therapy early in the course of the disease. However, standardization of the technique is necessary, and the correlation with clinical findings is of paramount importance due to the high sensitivity of this technique. PMID- 10457002 TI - [Prescription of drugs of systemic use by dentists]. AB - OBJECTIVE: The study of the prescription pattern of antibacterial and analgesic/antiinflammatory systemic medication by dentists. METHODS: Observational study based in questionnaires answered by a representative, randomly selected sample of 163 general dentists from the Metropolitan Region of Belo Horizonte, Southeastern Brazil. Some of the topics verified were: the most frequently prescribed drugs in the fortnight prior to the study, the use of generic names of drugs in the prescriptions, attendance at refresher courses on pharmacology, self-assessment of degree of knowledge on pharmacology, the importance given to this subject in the dentist's professional career, and the filling out of the clinical chart and the registration of the drugs prescribed on it. RESULTS: It was observed that the drugs were usually prescribed by their commercial name. There were a trend to prescribe more antiinflamatory than analgesics drugs. A small, but worrying, 13% of dentists didnot fill out the clinical chart for all the patients and 43% of the sample didnot register the drugs prescribed on it. The courses on pharmacology seemed to produce no significant alteration in self valuation as to the degree of knowledge in pharmacology and the use of the generic names of drugs. PMID- 10457003 TI - [Analysis of human anti-rabies post exposure treatment in the great area of Sao Paulo, Brazil]. AB - INTRODUCTION: The treatment of human beings with risk of exposure to rabies virus infection still presents some cases of undesirable post treatment reactions. The knowledge resulting from the regular analysis of the epidemiological features associated with these practices would minimize the occurrence of these troubles. METHODS: The records of the treatment of 8,758 human anti-rabies cases performed in Osasco City, S. Paulo, Brazil between 1984 and 1994 with Fuenzalida & Palacios vaccine and/or anti-rabies serum produced in equines were analysed by Epi Info program. RESULTS: It was found that the higher risk of exposure was for male children of from five to nine years of age. The most frequent scene of aggression was the victim's own house. Dogs were responsible for most of the accidents (62.8%); the majority of dogs and cats involved had not been immunized against rabies (51.0% of dogs and 73.2% of cats). Most people aged nine or under were attacked in the head (36.6%), followed by the upper limbs (35.1%); above this age the part of the body most frequently hurt was the upper (45.8%) and lower limbs (43. 7%); 26.5% of patients had already been vaccinated against rabies at least once in their lifetime; 90.7% of them sought medical care within five days, anti rabies treatment was prescribed for 41.9% of them. CONCLUSIONS: Post vaccinal reactions were observed in 0.25% of the patients 0.3% of which were of neurological; 80.1% of the dogs and 58.4% of the cats involved coere kept under observation. The months of March, July, August and September presented the highest aggression rates. PMID- 10457004 TI - [Experimental investigation of canine echinococcosis from swine hydatic cyst in Mexico]. AB - OBJECTIVE: To evaluate the infection and obtain the adult state of the cestode, echinococcosis was reproduced in dogs using the hydatic cyst of swine. METHODS: Two groups were formed, one of five and the other of three dogs, each animal in the experimental group was given two grams of germinative membrane of fertile hydatic cyst by oral route. The second was the control group. Both groups were evaluated clinically, serologically and parasitologically. One animal was killed on the 35th day after infection and each five successive days until the 55th day. In the second group all the animals were killed on the 55th day. Eggs of the cestode were observed in feces from the 51st post-infection day. The morphological evaluation was made through microscopic observation of the mucous intestine scraping. RESULTS: Fifty cestodes were analyzed, ten from each of the infected dogs, 49 (98%) presented three proglottids and 1 (2%) had four; 18 (36%) of the cestodes presented a gravid proglottid. The length of the strobila varied from 1.6 to 2.6 mm. The average of the long and short hooks was 31 and 34, respectively. The length of the long hooks varied from 0.081 to 0.09 mm, the short hooks from 0.034 to 0. 041 mm. The quantity of plasmatic proteins and the number of leukocytes were significantly greater in the control group (P < 0. 05); the quantity of alpha-globulins was larger in the infected group (P < 0.05). CONCLUSIONS: The results confirmed the dog-pig cycle, a subclinical infection in the definitive hosts, that makes the diagnosis and control in species closely related to the human being difficult. PMID- 10457005 TI - [Marking-release-recapture methods for determining the size of the natural population of Anopheles albitarsis l.s (Diptera:Culicidae)]. AB - OBJECTIVE: In order to evaluate the size of the Anopheles albitarsis s.l. population in the Ribeira Valley region of the south-eastern S. Paulo State, Brazil, marking-release-recapture experiments were carried out on the Experimental Farm in Pariquera-Acu county where an artificial irrigation system for the cultivation of rice has been installed. The experiments aimed at determining the vectorial capacity of this species. METHODS: During the summer (January, February and March), three marking-release-recapture experiments with field and laboratory reared populations were conducted to evaluate the population size. Simultaneous human bait collections were also carried out to estimate the human biting rate. RESULTS: The population sizes of the three experiments were estimated to be 64,560, 50,503 and 22,684 mosquitoes. The bite/man/night rate varied from 41.5 to 524,5. DISCUSSION: The results indicated high mosquito density in the summer which allow one to infer that, even though the survival rate is low, the number of remaining females is large enough to outlive the parasite's extrinsic developmental period. PMID- 10457006 TI - [Alternative models for the epidemiological analysis of obesity as a public health problem]. AB - Public health problems have been analyzed over time using different approaches involving a variety of methodological and theoretical models. The classical epidemiological model and the social epidemiological model are two examples of models used in public health research. Although each of these models covers a whole range of public research approaches, it is possible to identify the different theoretical and methodological criteria and the different ideological assumptions on which they are based. The objective of this article is to make a comparison of the criteria used by the classical and social epidemiological models in the study of public health problems, with special emphasis on the study of obesity. This comparison shows that the knowledge generated by studies carried out under each of these models is rather different, as an effect of the different theoretical, methodological and ideological criteria used by each model. However, far from being contradictory, the knowledge generated by each model can complement that generated by the other. Finally, the inclusion of social sciences as a tool for the analysis of public health problems, specifically the problem of obesity, is discussed. PMID- 10457008 TI - Regeneration of isthmic tissue is the result of a specific and direct interaction between rhombomere 1 and midbrain. AB - The midbrain-hindbrain boundary, or isthmus, is the source of signals that are responsible for regional specification of both the midbrain and anterior hindbrain. Fibroblast growth factor 8 (Fgf8) is expressed specifically at the isthmus and there is now good evidence that it forms at least part of the patterning signal. In this study, we use Fgf8 as a marker for isthmic cells to examine how interactions between midbrain and hindbrain can regenerate isthmic tissue and, thereby, gain insight into the normal formation and/or maintenance of the isthmus. We show that Fgf8-expressing tissue with properties of the isthmic organiser is generated when midbrain and rhombomere 1 tissue are juxtaposed but not when midbrain contacts any other rhombomere. The use of chick/quail chimeras shows that the isthmic tissue is largely derived from rhombomere 1. In a few cases a small proportion of the Fgf8-positive cells were of midbrain origin but this appears to be the result of a local respecification to a hindbrain phenotype, a process mimicked by ectopic FGF8. Studies in vitro show that the induction of Fgf8 is the result of a direct planar interaction between the two tissues and involves a diffusible signal. PMID- 10457007 TI - Zebrafish narrowminded suggests a genetic link between formation of neural crest and primary sensory neurons. AB - In the developing vertebrate nervous system, both neural crest and sensory neurons form at the boundary between non-neural ectoderm and the neural plate. From an in situ hybridization based expression analysis screen, we have identified a novel zebrafish mutation, narrowminded (nrd), which reduces the number of early neural crest cells and eliminates Rohon-Beard (RB) sensory neurons. Mosaic analysis has shown that the mutation acts cell autonomously suggesting that nrd is involved in either the reception or interpretation of signals at the lateral neural plate boundary. Characterization of the mutant phenotype indicates that nrd is required for a primary wave of neural crest cell formation during which progenitors generate both RB sensory neurons and neural crest cells. Moreover, the early deficit in neural crest cells in nrd homozygotes is compensated later in development. Thus, we propose that a later wave can compensate for the loss of early neural crest cells but, interestingly, not the RB sensory neurons. We discuss the implications of these findings for the possibility that RB sensory neurons and neural crest cells share a common evolutionary origin. PMID- 10457009 TI - Functions for Drosophila brachyenteron and forkhead in mesoderm specification and cell signalling. AB - The visceral musculature of the larval midgut of Drosophila has a lattice-type structure and consists of an inner stratum of circular fibers and an outer stratum of longitudinal fibers. The longitudinal fibers originate from the posterior tip of the mesoderm anlage, which has been termed the caudal visceral mesoderm (CVM). In this study, we investigate the specification of the CVM and particularly the role of the Drosophila Brachyury-homologue brachyenteron. Supported by fork head, brachyenteron mediates the early specification of the CVM along with zinc-finger homeodomain protein-1. This is the first function described for brachyenteron or fork head in the mesoderm of Drosophila. The mode of cooperation resembles the interaction of the Xenopus homologues Xbra and Pintallavis. Another function of brachyenteron is to establish the surface properties of the CVM cells, which are essential for their orderly migration along the trunk-derived visceral mesoderm. During this movement, the CVM cells, under the control of brachyenteron, induce the formation of one muscle/pericardial precursor cell in each parasegment. We propose that the functions of brachyenteron in mesodermal development of Drosophila are comparable to the roles of the vertebrate Brachyury genes during gastrulation. PMID- 10457010 TI - Bmp signaling regulates proximal-distal differentiation of endoderm in mouse lung development. AB - In the mature mouse lung, the proximal-distal (P-D) axis is delineated by two distinct epithelial subpopulations: the proximal bronchiolar epithelium and the distal respiratory epithelium. Little is known about the signaling molecules that pattern the lung along the P-D axis. One candidate is Bone Morphogenetic Protein 4 (Bmp4), which is expressed in a dynamic pattern in the epithelial cells in the tips of growing lung buds. Previous studies in which Bmp4 was overexpressed in the lung endoderm (Bellusci, S., Henderson, R., Winnier, G., Oikawa, T. and Hogan, B. L. M. (1996) Development 122, 1693-1702) suggested that this factor plays an important role in lung morphogenesis. To further investigate this question, two complementary approaches were utilized to inhibit Bmp signaling in vivo. The Bmp antagonist Xnoggin and, independently, a dominant negative Bmp receptor (dnAlk6), were overexpressed using the surfactant protein C (Sp-C) promoter/enhancer. Inhibiting Bmp signaling results in a severe reduction in distal epithelial cell types and a concurrent increase in proximal cell types, as indicated by morphology and expression of marker genes, including the proximally expressed hepatocyte nuclear factor/forkhead homologue 4 (Hfh4) and Clara cell marker CC10, and the distal marker Sp-C. In addition, electron microscopy demonstrates the presence of ciliated cells, a proximal cell type, in the most peripheral regions of the transgenic lungs. We propose a model in which Bmp4 is a component of an apical signaling center controlling P-D patterning. Endodermal cells at the periphery of the lung, which are exposed to high levels of Bmp4, maintain or adopt a distal character, while cells receiving little or no Bmp4 signal initiate a proximal differentiation program. PMID- 10457011 TI - Nurr1, an orphan nuclear receptor, is a transcriptional activator of endogenous tyrosine hydroxylase in neural progenitor cells derived from the adult brain. AB - Adult rat-derived hippocampal progenitor cells express many of the molecules implicated in midbrain dopaminergic determination, including FGF receptors 1, 2 and 3, the sonic hedgehog receptor components Smo and Ptc, and the region specific transcription factors Ptx3 and Nurr1. Here we use undifferentiated progenitors to probe the events leading to the dopaminergic phenotype and find that the influences of Nurr1 can be temporally and mechanistically uncoupled from the patterning influences of sonic hedgehog and FGF-8 or the more generic process of neuronal differentiation itself. In gain-of-function experiments, Nurr1 is able to activate transcription of the tyrosine hydroxylase gene by binding a response element within a region of the tyrosine hydroxylase promoter necessary for midbrain-specific expression. This activation is mediated through a retinoid X receptor independent mechanism and occurs in all precursors, regardless of differentiation status. Overexpression of Nurr1 does not affect proliferation or stimulate neuronal differentiation and has no influence on the expression of other dopaminergic markers. This uncoupling of tyrosine hydroxylase expression from other dopaminergic markers suggests that the midbrain dopaminergic identity is dictated by a combination of pan-dopaminergic (e.g., Shh/FGF-8) and region specific (Nurr1) mechanisms. PMID- 10457012 TI - Loss of p27Kip1 function results in increased proliferative capacity of oligodendrocyte progenitors but unaltered timing of differentiation. AB - In many tissues, progenitor cells permanently withdraw from the cell cycle prior to commitment towards a differentiated phenotype. In the oligodendrocyte lineage a counting mechanism has been proposed, linking the number of cell divisions to growth arrest and differentiation. A direct prediction of this model is that an increase in the number of cell divisions would result in a delayed onset of differentiation. Since the cell cycle inhibitor p27Kip1 is an essential component of the machinery leading to oligodendrocyte progenitor growth arrest, we examined the temporal relationship between cell cycle withdrawal and expression of late differentiation markers in vivo, in mice carrying a targeted deletion in the p27Kip1 gene. Using bromodeoxyuridine to label proliferating cells, quaking (QKI) to identify embryonic glial progenitors, NG2 to identify neonatal oligodendrocyte progenitors, and myelin basic protein to label differentiated oligodendrocytes, we found an increased number of proliferating QKI- and NG2-positive cells in germinal zones of p27Kip1(-/-) mice at the peak of gliogenesis. However, no delay was observed in these mice in the appearance of the late differentiation marker myelin basic protein in the developing corpus callosum and cerebellum. Significantly, a decrease in cyclin E levels was observed in the brain of p27Kip1 null mice coincident with oligodendrocyte growth arrest. We conclude that two distinct modalities of growth arrest occur in the oligodendrocyte lineage: a p27Kip1-dependent mechanism of growth arrest affecting proliferation in early phases of gliogenesis, and a p27Kip1-independent event leading to withdrawal from the cell cycle and differentiation. PMID- 10457013 TI - Differential requirements of the fused kinase for hedgehog signalling in the Drosophila embryo. AB - The two signalling proteins, Wingless and Hedgehog, play fundamental roles in patterning cells within each metamere of the Drosophila embryo. Within the ventral ectoderm, Hedgehog signals both to the anterior and posterior directions: anterior flanking cells express the wingless and patched Hedgehog target genes whereas posterior flanking cells express only patched. Furthermore, Hedgehog acts as a morphogen to pattern the dorsal cuticle, on the posterior side of cells where it is produced. Thus responsive embryonic cells appear to react according to their position relative to the Hedgehog source. The molecular basis of these differences is still largely unknown. In this paper we show that one component of the Hedgehog pathway, the Fused kinase accumulates preferentially in cells that could respond to Hedgehog but that Fused concentration is not a limiting step in the Hedgehog signalling. We present direct evidence that Fused is required autonomously in anterior cells neighbouring Hedgehog in order to maintain patched and wingless expression while Wingless is in turn maintaining engrailed and hedgehog expression. By expressing different components of the Hedgehog pathway only in anterior, wingless-expressing cells we could show that the Hedgehog signalling components Smoothened and Cubitus interruptus are required in cells posterior to Hedgehog domain to maintain patched expression whereas Fused is not necessary in these cells. This result suggests that Hedgehog responsive ventral cells in embryos can be divided into two distinct types depending on their requirement for Fused activity. In addition, we show that the morphogen Hedgehog can pattern the dorsal cuticle independently of Fused. In order to account for these differences in Fused requirements, we propose the existence of position specific modulators of the Hedgehog response. PMID- 10457014 TI - Sonic hedgehog controls epaxial muscle determination through Myf5 activation. AB - Sonic hedgehog (Shh), produced by the notochord and floor plate, is proposed to function as an inductive and trophic signal that controls somite and neural tube patterning and differentiation. To investigate Shh functions during somite myogenesis in the mouse embryo, we have analyzed the expression of the myogenic determination genes, Myf5 and MyoD, and other regulatory genes in somites of Shh null embryos and in explants of presomitic mesoderm from wild-type and Myf5 null embryos. Our findings establish that Shh has an essential inductive function in the early activation of the myogenic determination genes, Myf5 and MyoD, in the epaxial somite cells that give rise to the progenitors of the deep back muscles. Shh is not required for the activation of Myf5 and MyoD at any of the other sites of myogenesis in the mouse embryo, including the hypaxial dermomyotomal cells that give rise to the abdominal and body wall muscles, or the myogenic progenitor cells that form the limb and head muscles. Shh also functions in somites to establish and maintain the medio-lateral boundaries of epaxial and hypaxial gene expression. Myf5, and not MyoD, is the target of Shh signaling in the epaxial dermomyotome, as MyoD activation by recombinant Shh protein in presomitic mesoderm explants is defective in Myf5 null embryos. In further support of the inductive function of Shh in epaxial myogenesis, we show that Shh is not essential for the survival or the proliferation of epaxial myogenic progenitors. However, Shh is required specifically for the survival of sclerotomal cells in the ventral somite as well as for the survival of ventral and dorsal neural tube cells. We conclude, therefore, that Shh has multiple functions in the somite, including inductive functions in the activation of Myf5, leading to the determination of epaxial dermomyotomal cells to myogenesis, as well as trophic functions in the maintenance of cell survival in the sclerotome and adjacent neural tube. PMID- 10457015 TI - Development of the Drosophila mushroom bodies: sequential generation of three distinct types of neurons from a neuroblast. AB - The mushroom bodies (MBs) are prominent structures in the Drosophila brain that are essential for olfactory learning and memory. Characterization of the development and projection patterns of individual MB neurons will be important for elucidating their functions. Using mosaic analysis with a repressible cell marker (Lee, T. and Luo, L. (1999) Neuron 22, 451-461), we have positively marked the axons and dendrites of multicellular and single-cell mushroom body clones at specific developmental stages. Systematic clonal analysis demonstrates that a single mushroom body neuroblast sequentially generates at least three types of morphologically distinct neurons. Neurons projecting into the (gamma) lobe of the adult MB are born first, prior to the mid-3rd instar larval stage. Neurons projecting into the alpha' and beta' lobes are born between the mid-3rd instar larval stage and puparium formation. Finally, neurons projecting into the alpha and beta lobes are born after puparium formation. Visualization of individual MB neurons has also revealed how different neurons acquire their characteristic axon projections. During the larval stage, axons of all MB neurons bifurcate into both the dorsal and medial lobes. Shortly after puparium formation, larval MB neurons are selectively pruned according to birthdays. Degeneration of axon branches makes early-born gamma neurons retain only their main processes in the peduncle, which then project into the adult gamma lobe without bifurcation. In contrast, the basic axon projections of the later-born (alpha'/beta') larval neurons are preserved during metamorphosis. This study illustrates the cellular organization of mushroom bodies and the development of different MB neurons at the single cell level. It allows for future studies on the molecular mechanisms of mushroom body development. PMID- 10457016 TI - A head-activator binding protein is present in hydra in a soluble and a membrane anchored form. AB - The neuropeptide head activator plays an important role for proliferation and determination of stem cells in hydra. By affinity chromatography a 200 kDa head activator binding protein, HAB, was isolated from the multiheaded mutant of Chlorohydra viridissima. Partial amino acid sequences were used to clone the HAB cDNA which coded for a receptor with a unique alignment of extracellular modules, a transmembrane domain, and a short carboxy-terminal cytoplasmic tail. A mammalian HAB homologue with identical alignment of these modules is expressed early in brain development. Specific antibodies revealed the presence of HAB in hydra as a transmembrane receptor, but also as secreted protein, both capable of binding head activator. Secretion of HAB during regeneration and expression in regions of high determination potential hint at a role for HAB in regulating the concentration and range of action of head activator. PMID- 10457017 TI - The Phox2 homeodomain proteins are sufficient to promote the development of sympathetic neurons. AB - The development of sympathetic neurons is controlled by a network of transcriptional regulators, including the paired homeodomain proteins Phox2a and Phox2b. To understand the role of Phox2 proteins in more detail, the effect of Phox2 overexpression was analysed in the avian peripheral nervous system. Phox2a expression in neural crest cultures elicited a strong increase in the number of sympathoadrenergic cells. Expression of Phox2a in the chick embryo promoted the generation of additional neurons expressing the noradrenergic marker genes DBH and TH, pan-neuronal genes SCG10 and NF160 and cholinergic genes ChAT and VAChT. Phox2a-induced neurons were found in ectopic locations such as dorsal root ganglia and peripheral nerve. Sympathoadrenergic development could be elicited in cultures of E5 dorsal root ganglia, demonstrating the presence of Phox2a responsive cells in non-autonomic peripheral ganglia. Phox2b induced ectopic neurons in the chick embryo in the same way as Phox2a. These results show that Phox2 proteins are sufficient to promote sympathetic neuron generation and control, directly or indirectly, the expression of a large number of genes characteristic for sympathetic neurons. PMID- 10457018 TI - Successive patterns of clonal cell dispersion in relation to neuromeric subdivision in the mouse neuroepithelium. AB - We made use of the laacz procedure of single-cell labelling to visualize clones labelled before neuromere formation, in 12.5-day mouse embryos. This allowed us to deduce two successive phases of cell dispersion in the formation of the rhombencephalon: an initial anterior-posterior (AP) cell dispersion, followed by an asymmetrical dorsoventral (DV) cell distribution during which AP cell dispersion occurs in territories smaller than one rhombomere. We conclude that the general arrest of AP cell dispersion precedes the onset of morphological segmentation and is not imposed by the interface between adjacent rhombomeres. This demonstrates a major change in the mode of epithelial growth that precedes or accompanies the formation of neuromeres. We also deduced that the period of DV cell dispersion in the neuroepithelium is followed by a coherent growth phase. These results suggest a cell organization on a Cartesian grid, the coordinates of which correspond to the AP and DV axis of the neural tube. A similar sequence of AP cell dispersion followed by an arrest of AP cell dispersion, a preferential DV cell dispersion and then by a coherent neuroepithelial growth, is also observed in the spinal cord and mesencephalon. This demonstrates that a similar cascade of cell events occurs in these different domains of the CNS. In the prosencephalon, differences in spatial constraints may explain the variability in the orientation of cell clusters. Genetic and clonal patterning in the AP and DV dimensions follow the same spatial sequence. An interesting possibility is that these successive patterns of cell growth facilitate the acquisition of positional information. PMID- 10457020 TI - Members of the YABBY gene family specify abaxial cell fate in Arabidopsis. AB - Lateral organs produced by shoot apical and flower meristems exhibit a fundamental abaxial-adaxial asymmetry. We describe three members of the YABBY gene family, FILAMENTOUS FLOWER, YABBY2 and YABBY3, isolated on the basis of homology to CRABS CLAW. Each of these genes is expressed in a polar manner in all lateral organ primordia produced from the apical and flower meristems. The expression of these genes is precisely correlated with abaxial cell fate in mutants in which abaxial cell fates are found ectopically, reduced or eliminated. Ectopic expression of either FILAMENTOUS FLOWER or YABBY3 is sufficient to specify the development of ectopic abaxial tissues in lateral organs. Conversely, loss of polar expression of these two genes results in a loss of polar differentiation of tissues in lateral organs. Taken together, these observations indicate that members of this gene family are responsible for the specification of abaxial cell fate in lateral organs of Arabidopsis. Furthermore, ectopic expression studies suggest that ubiquitous abaxial cell fate and maintenance of a functional apical meristem are incompatible. PMID- 10457019 TI - Mutual antagonism between signals secreted by adjacent wingless and engrailed cells leads to specification of complementary regions of the Drosophila parasegment. AB - Specialized groups of cells known as organizers govern the establishment of cell type diversity across cellular fields. Segmental patterning within the Drosophila embryonic epidermis is one paradigm for organizer function. Here cells differentiate into smooth cuticle or distinct denticle types. At parasegment boundaries, cells expressing Wingless confront cells co-expressing Engrailed and Hedgehog. While Wingless is essential for smooth cell fates, the signals that establish denticle diversity are unknown. We show that wg mutants have residual mirror-symmetric pattern that is due to an Engrailed-dependent signal specifying anterior denticle fates. The Engrailed-dependent signal acts unidirectionally and Wg activity imposes this asymmetry. Reciprocally, the Engrailed/Hedgehog interface imposes asymmetry on Wg signaling. Thus, a bipartite organizer, with each signal acting essentially unidirectionally, specifies segmental pattern. PMID- 10457021 TI - Coelom formation: binary decision of the lateral plate mesoderm is controlled by the ectoderm. AB - Most triploblastic animals including vertebrates have a coelomic cavity that separates the outer and inner components of the body. The coelom is lined by two different tissue components, somatopleure and splanchnopleure, which are derived from the lateral plate region. Thus, the coelom is constructed as a result of a binary decision during early specification of the lateral plate. In this report we studied the molecular mechanisms of this binary decision. We first demonstrate that the splitting of the lateral plate into the two cell sheets progresses in an anteroposterior order and this progression is not coordinated with that of the somitic segmentation. By a series of embryological manipulations we found that young splanchnic mesoderm is still competent to be respecified as somatic mesoderm, and the ectoderm overlying the lateral plate is sufficient for this redirection. The lateral ectoderm is also required for maintenance of the somatic character of the mesoderm. Thus, the ectoderm plays at least two roles in the early subdivision of the lateral plate: specification and maintenance of the somatic mesoderm. We also show that the latter interactions are mediated by BMP molecules that are localized in the lateral ectoderm. Evolutionary aspects of the coelom formation are also considered. PMID- 10457022 TI - Sprouty is a general inhibitor of receptor tyrosine kinase signaling. AB - Sprouty was originally identified as an inhibitor of Drosophila FGF receptor signaling during tracheal development. By following the capacity of ectopic Sprouty to abolish the pattern of activated MAP kinase in embryos, we show that Sprouty can inhibit other receptor tyrosine kinase (RTK) signaling pathways, namely the Heartless FGF receptor and the EGF receptor. Similarly, in wing imaginal discs, ectopic Sprouty abolishes activated MAP kinase induced by the EGF receptor pathway. Sprouty expression is induced by the EGFR pathway in some, but not all, tissues in which EGFR is activated, most notably in follicle cells of the ovary, the wing imaginal disc and the eye disc. In the ovary, induction of sprouty expression follows the pattern of EGFR activation in the follicle cells. Generation of homozygous sprouty mutant follicle-cell clones demonstrates an essential role for Sprouty in restricting EGFR activation throughout oogenesis. At the stage when dorso-ventral polarity of the follicle cells is established, Sprouty limits the ventral expansion of the activating Gurken signal. Later, when dorsal appendage fates are determined, reduction of signaling by Sprouty facilitates the induction of inter-appendage cell fates. The capacity of Sprouty to reduce or eliminate accumulation of activated MAP kinase indicates that in vivo it intersects with the pathway upstream to MAP kinase. The ability of ectopic Sprouty to rescue lethality caused by activated Raf suggests that it may impinge upon the pathway by interacting with Raf or downstream to it. PMID- 10457023 TI - Targeted ablation of secretin-producing cells in transgenic mice reveals a common differentiation pathway with multiple enteroendocrine cell lineages in the small intestine. AB - The four cell types of gut epithelium, enteroendocrine cells, enterocytes, Paneth cells and goblet cells, arise from a common totipotent stem cell located in the mid portion of the intestinal gland. The secretin-producing (S) cell is one of at least ten cell types belonging to the diffuse neuroendocrine system of the gut. We have examined the developmental relationship between secretin cells and other enteroendocrine cell types by conditional ablation of secretin cells in transgenic mice expressing herpes simplex virus 1 thymidine kinase (HSVTK). Ganciclovir-treated mice showed markedly increased numbers of apoptotic cells at the crypt-villus junction. Unexpectedly, ganciclovir treatment induced nearly complete ablation of enteroendocrine cells expressing cholecystokinin and peptide YY/glucagon (L cells) as well as secretin cells, suggesting a close developmental relationship between these three cell types. In addition, ganciclovir reduced the number of enteroendocrine cells producing gastric inhibitory polypeptide, substance-P, somatostatin and serotonin. During recovery from ganciclovir treatment, the enteroendocrine cells repopulated the intestine in normal numbers, suggesting that a common early endocrine progenitor was spared. Expression of BETA2, a basic helix-loop-helix protein essential for differentiation of secretin and cholecystokinin cells was examined in the proximal small intestine. BETA2 expression was seen in all enteroendocrine cells and not seen in nonendocrine cells. These results suggest that most small intestinal endocrine cells are developmentally related and that a close developmental relationship exists between secretin-producing S cells and cholecystokinin-producing and L type enteroendocrine cells. In addition, our work shows the existence of a multipotent endocrine-committed cell type and locates this hybrid multipotent cell type to a region of the intestine populated by relatively immature cells. PMID- 10457024 TI - Regulation of neuronal K(+) currents by target-derived factors: opposing actions of two different isoforms of TGFbeta. AB - The developmental expression of macroscopic Ca(2+)-activated K(+) currents in chick ciliary ganglion neurons is dependent on an avian ortholog of TGFbeta1, known as TGFbeta4, secreted from target tissues in the eye. Here we report that a different isoform, TGFbeta3, is also expressed in a target tissue of ciliary ganglion neurons. Application of TGFbeta3 inhibits the functional expression of whole-cell Ca(2+)-activated K(+) currents evoked by 12 hour treatment with either TGFbeta1 or beta-neuregulin-1 in ciliary ganglion neurons developing in vitro. TGFbeta3 had no effect on voltage-activated Ca(2+) currents. A neutralizing antiserum specific for TGFbeta3 potentiates stimulation of Ca(2+)-activated K(+) currents evoked by a target tissue (iris) extract in cultured ciliary ganglion neurons, indicating that TGFbeta3 is an inhibitory component of these extracts. Intraocular injection of TGFbeta3 causes a modest but significant inhibition of the expression of Ca(2+)-activated K(+) currents in ciliary ganglion neurons developing in vivo. Further, intraocular injection of a TGFbeta3-neutralizing antiserum stimulates expression of Ca(2+)-activated K(+) currents in ciliary ganglion neurons developing in vivo, indicating that endogenous TGFbeta3 regulates the functional expression of this current. The normal developmental expression of functional Ca(2+)-activated K(+) currents in ciliary ganglion neurons developing in vivo is therefore regulated by two different target-derived isoforms of TGFbeta, which produce opposing effects on the electrophysiological differentiation of these neurons. PMID- 10457025 TI - A Drosophila Axin homolog, Daxin, inhibits Wnt signaling. AB - The vertebrate Axin protein, the product of the mouse fused gene, binds to beta catenin to inhibit Wnt signaling. We have identified a homolog of Axin in Drosophila, Daxin. Using double-stranded RNA interference, we generated loss-of function phenotypes that are similar to overexpression of the Drosophila Wnt gene wingless (wg). Overexpression of Daxin produces phenotypes similar to loss of wg. In addition, we show that Daxin overexpression can modify phenotypes elicited by wg and another Drosophila Wnt gene, DWnt-2. Using immunoprecipitation of endogenous Daxin protein from embryos we show that Daxin interacts with Armadillo and Zeste-white 3. The loss-of-function and overexpression phenotypes show that Daxin, like its mammalian counterpart, acts as a negative regulator of wg/Wnt signaling. PMID- 10457026 TI - Frizzled and Dfrizzled-2 function as redundant receptors for Wingless during Drosophila embryonic development. AB - In cell culture assays, Frizzled and Dfrizzled2, two members of the Frizzled family of integral membrane proteins, are able to bind Wingless and transduce the Wingless signal. To address the role of these proteins in the intact organism and to explore the question of specificity of ligand-receptor interactions in vivo, we have conducted a genetic analysis of frizzled and Dfrizzled2 in the embryo. These experiments utilize a small gamma-ray-induced deficiency that uncovers Dfrizzled2. Mutants lacking maternal frizzled and zygotic frizzled and Dfrizzled2 exhibit defects in the embryonic epidermis, CNS, heart and midgut that are indistinguishable from those observed in wingless mutants. Epidermal patterning defects in the frizzled, Dfrizzled2 double-mutant embryos can be rescued by ectopic expression of either gene. In frizzled, Dfrizzled2 mutant embryos, ectopic production of Wingless does not detectably alter the epidermal patterning defect, but ectopic production of an activated form of Armadillo produces a naked cuticle phenotype indistinguishable from that produced by ectopic production of activated Armadillo in wild-type embryos. These experiments indicate that frizzled and Dfrizzled2 function downstream of wingless and upstream of armadillo, consistent with their proposed roles as Wingless receptors. The lack of an effect on epidermal patterning of ectopic Wingless in a frizzled, Dfrizzled2 double mutant argues against the existence of additional Wingless receptors in the embryo or a model in which Frizzled and Dfrizzled2 act simply to present the ligand to its bona fide receptor. These data lead to the conclusion that Frizzled and Dfrizzled2 function as redundant Wingless receptors in multiple embryonic tissues and that this role is accurately reflected in tissue culture experiments. The redundancy of Frizzled and Dfrizzled2 explains why Wingless receptors were not identified in earlier genetic screens for mutants defective in embryonic patterning. PMID- 10457027 TI - Are all helicobacters equal? Mechanisms of gastroduodenal pathology and their clinical implications. AB - Most cases of peptic ulcer disease, gastric mucosa associated lymphoid tissue (MALT) lymphoma and cancer of the distal stomach are complications of Helicobacter pylori infection. However, as with most infections not all patients who contract the infection develop the complications of the disease. The other factors that influence the likelihood of problems arising are the virulence of the infecting organism, the genetic constitution and age of the host, and environmental factors. This paper focuses mainly upon the effect of strain differences and the causation of serious disease. There is considerable genetic variation between the different strains of H pylori, some causing a more severe inflammatory response in the host than others. These strains are also associated with a greater likelihood of causing peptic ulcer, atrophic gastritis and intestinal metaplasia and gastric cancer. There is some evidence to suggest that these more virulent organisms may also protect the host from the development of reflux oesophagitis and possibly cancer in the region of the gastro-oesophageal junction. The major difference between virulent and relatively avirulent organisms depends upon the presence of the cag pathogenicity island, a segment of DNA that has been acquired possibly from another organism and is now incorporated within the helicobacter genome. Its presence is associated with the secretion of the vacuolating toxin which is a protein known to cause damage in cell culture and in vivo. As CagA, one of the proteins produced by the pathogenicity island, is highly antigenic, people infected with more virulent strains can be identified by a blood test. Currently controversy surrounds the question as to whether all patients with H pylori should be treated for infection or whether medication should be reserved for those who already have the complications of the infection, or individuals infected with the more virulent strain of the organism. PMID- 10457028 TI - Gastric precancerous lesions: heading for an international consensus. AB - As pathological criteria lie at the foundation for the classification of many diseases, a crucial requisite for such classifications to be valid is that their morphological basis be standardised. Inadequately standardised diagnostic criteria result in unacceptable interobserver variation, a factor that may influence both individual patient care and the evaluation of clinical protocols. One of the most important goals in gastric diseases today is to establish whether cure of Helicobacter pylori is an effective preventive measure against gastric cancer. To tackle this issue it is necessary to measure reliably intermediate outcomes, specifically gastric atrophy and dysplasia. However, there is little agreement on what gastric atrophy and atrophic gastritis are, and treatment and follow up results obtained at one clinical centre are often radically different from those obtained at another. Similarly, studies that examine the fate of dysplastic lesions in the stomach show a great divergence of outcomes between Europe and North America and Japan, where the concept of dysplasia has different connotations. To reach a consensus on the definitions and diagnostic criteria for atrophy and atrophic gastritis, a group of gastrointestinal pathologists and gastroenterologists met in Houston, Texas, USA, in February 1998. Substantive progress was made, but several problems remained, and a study aimed at resolving the issues that seem to stand in the way of an international agreement is currently underway. To fulfil the need for a broad discussion on the diagnostic differences of gastric dysplasia and cancer between East and West, an international group of pathologists gathered in Padova, Italy, in the spring of 1998. Their main objectives were: (1) to agree on the definitions of the spectrum of gastric preneoplastic lesions; (2) to establish an international glossary for gastric precancerous lesions; and (3) to test the consensus and eventually generate guidelines useful to clinicians for the development of management strategies. A consensus was achieved on the definition of gastric dysplasia as preinvasive neoplasia. Other validation studies are underway. The experiences achieved in the search for an international consensus on the phenotypes of atrophic gastritis and gastric dysplasia may represent a model in dealing with the new scenario of a modern evidence-based pathology. PMID- 10457029 TI - Extradigestive manifestations of Helicobacter pylori gastric infection. AB - In the past year, several studies have been carried out on the association between Helicobacter pylori infection and a miscellany of extradigestive diseases, such as cardiovascular, immunological, and various other pathologies. In particular, a higher prevalence of H pylori infection in patients affected by ischaemic heart disease has been described and there is growing evidence for an association between H pylori and some autoimmune diseases. Moreover, recent studies have shown that various helicobacter species have been detected in human bile; if confirmed, this finding could revise the diagnostic and therapeutic approach to diseases of the biliary tract. PMID- 10457030 TI - Gastro-oesophageal reflux disease and Helicobacter pylori: an intricate relation. AB - Heartburn is a common symptom affecting 21-44% of the adult population on a monthly basis. Oesophagitis is less common, affecting 2% of individuals. Epidemiological studies have shown that patients with gastro-oesophageal reflux disease (GORD) have similar incidence rates of Helicobacter pylori infection as do controls. Some groups have reported that there is a lower incidence, deducing that infection does not cause, and in some way confers protection against GORD. Additional supportive evidence is available from reports of GORD development following successful H pylori eradication. The mechanisms involved are complicated. Individuals with H pylori induced pangastritis and subsequent hypochlorhydria may be protected whereas those with an antral predominant gastritis, as in duodenal ulcer disease, with an increased acid output may be prone to development of GORD. Recent evidence has linked H pylori infection with the development of inflammation of the gastric cardia---carditis. Reports are available which show that carditis is a frequent finding in patients with GORD. The incidence of both cardia and oesophageal carcinoma is increasing. The relation between GORD, carditis, intestinal metaplasia, and cardia carcinoma is unclear. Intestinal metaplasia may result from multifocal atrophic gastritis, linked to H pylori infection or from GORD and the development of Barrett's oesophagus. Long term follow up studies will be required to assess the malignant potential of these histological entities and whether or not H pylori infection has an aetiological role. PMID- 10457031 TI - The 13C urea breath test in the diagnosis of Helicobacter pylori infection. AB - The urea breath test (UBT) is one of the most important non-invasive methods for detecting Helicobacter pylori infection. The test exploits the hydrolysis of orally administered urea by the enzyme urease, which H pylori produces in large quantities. Urea is hydrolysed to ammonia and carbon dioxide, which diffuses into the blood and is excreted by the lungs. Isotopically labelled CO2 can be detected in breath using various methods. Labelling urea with 13C is becoming increasingly popular because this non-radioactive isotope is innocuous and can be safely used in children and women of childbearing age. Breath samples can also be sent by post or courier to remote analysis centres. The test is easy to perform and can be repeated as often as required in the same patient. A meal must be given to increase the contact time between the tracer and the H pylori urease inside the stomach. The test has been simplified to the point that two breath samples collected before and 30 minutes after the ingestion of urea in a liquid form suffice to provide reliable diagnostic information. The cost of producing 13C urea is high, but it may be possible to reduce the dosage further by administering it in capsule form. An isotope ratio mass spectrometer (IRMS) is generally used to measure 13C enrichment in breath samples, but this machine is expensive. In order to reduce this cost, new and cheaper equipment based on non dispersive, isotope selective, infrared spectroscopy (NDIRS) and laser assisted ratio analysis (LARA) have recently been developed. These are valid alternatives to IRMS although they cannot process the same large number of breath samples simultaneously. These promising advances will certainly promote the wider use of the 13C-UBT, which is especially useful for epidemiological studies in children and adults, for screening patients before endoscopy, and for assessing the efficacy of eradication regimens. PMID- 10457032 TI - New immunological assays for the diagnosis of Helicobacter pylori infection. AB - There are several types of immunological tests available for the diagnosis and management of Helicobacter pylori infection. Most commercially available serological kits use the enzyme linked immunosorbent assay (ELISA) test format. Originally the kits used crude antigen preparations although many of the newer kits use a more purified antigen preparation, with often increased specificity but lower sensitivity. Near patient test kits are based either on latex agglutination or immunochromatography. Generally they have low sensitivities compared with laboratory tests. Western blotting, ELISA, and recombinant immunoblot assays (RIBA) have also been developed into commercially available kits and can be used to indicate the presence of specific virulence markers. An antigen detection kit has been developed for the detection of Helicobacter pylori in faeces. Immunological reagents have also been combined with other diagnostic modalities to develop immunohistochemical stains and DNA immunoassays. Helicobacter pylori is now recognised as the cause of gastritis and most cases of peptic ulcer disease (PUD); its long term carriage increases the risk of gastric adenocarcinoma sixfold and it is designated as a class I carcinogen. H pylori has also been implicated as a cause of gastric mucosa associated lymphoid tissue lymphomas. Its relation to non-ulcer dyspepsia remains controversial. Additionally, long term carriage of the organism may be associated with short stature in young girls and, in the general population, as a possible risk factor for the development of vasospastic disorders and possibly skin immunopathology such as urticaria. With the recognition of H pylori as an important human pathogen, it has become one of the growing number of organisms to have its complete genome sequence mapped. Serology is an important method of determining colonisation status and can be used for diagnosis, as a screening procedure, or to follow the efficacy of eradication regimens. Most serological assays are in the ELISA format although some are based on the latex agglutination reaction. These latter are used principally as near patient assays. Most assays detect IgG in serum although some detect serum IgA. More recently developed assays detect IgA in saliva and the production of affinity purified antibodies has led to the development of an antigen detection assay for faecal specimens. Serological reagents have also been used in immunocytochemistry and to speed up the detection of amplified products of the polymerase chain reaction (PCR)-DNA immunoassays. PMID- 10457033 TI - How should Helicobacter pylori positive dyspeptic patients be managed? AB - Dyspepsia, according to the internationally accepted Rome criteria, refers to pain or discomfort centred in the upper abdomen; patients with predominant heartburn are excluded from this group, although minor or infrequent heartburn is commonly associated with dyspepsia. It is an important condition not only because it is common and costly, but because it may indicate the presence of serious disease such as peptic ulcer or gastric cancer. However, the most frequent causes of dyspepsia are functional dyspepsia and gastro-oesophageal reflux disease. The discovery of Helicobacter pylori has resulted in important advances in the management of dyspepsia. The clinician faced with a patient who has persistent or recurrent dyspepsia needs to differentiate clearly those patients who have not been previously investigated from patients documented to have functional dyspepsia after investigation (fig 1). Here, the management of H pylori positive dyspeptic patients who have and have not been fully investigated will be reviewed. PMID- 10457034 TI - How should Helicobacter pylori negative patients be managed? AB - Dyspepsia is a digestive syndrome distinct from (although frequently overlapping with) gastro-oesophageal reflux disease (GORD) and irritable bowel syndrome (IBS), which is characterised by various combinations of painful and non-painful symptoms arising from the epigastrium. Dyspepsia can be secondary to a variety of diseases, but in most instances it is idiopathic. Helicobacter pylori infection is responsible for the majority of peptic ulcers and of other diseases potentially associated with dyspepsia. Nevertheless, a causal role for H pylori infection in symptom occurrence has not been established. Experimental data indicate that H pylori eradication does not improve symptoms in the majority of dyspeptic patients. It has been proposed recently that H pylori negative patients should be managed according to their clinical presentation. Some reports suggest that taking into consideration the most relevant or "predominant" symptom may help to identify distinct subgroups among dyspeptic patients with different underlying pathophysiological abnormalities and different responses to treatment. Well designed and conducted prospective studies are needed to verify whether treatment of H pylori negative dyspeptic patients based on the predominant symptom actually is a cost effective approach. PMID- 10457035 TI - How should Helicobacter pylori infected children be managed? AB - It is now recognised that Helicobacter pylori, like most enteric infections, is mainly acquired in childhood. Adults rarely become infected, with seroconversion rates varying between 0.33and 0.5% per person year. The age at which children are most likely to become infected is still unclear, but findings in a number of cross-sectional studies suggest that infection is acquired before the age of five. The prevalence of infection is highest in children in the developing world where up to 75% of children may be infected by the age of 10. In the developed world the prevalence of infection is noticeably increased among socially deprived children. The diagnosis of H pylori infection in childhood is most often made at endoscopy, for which there are many indications. Symptoms such as abdominal pain, vomiting, and haematemesis may be associated with duodenal ulcer and H pylori infection. However, in the case of children undergoing endoscopy for assessment of oesophagitis, failure to thrive, coeliac disease, Crohn's disease, or portal hypertension, the finding of H pylori infection is likely to be incidental. How should we manage these children with a diagnosis of H pylori infection? Currently, there are no consensus guidelines for the management of H pylori infected children. In 1994 the National Institutes of Health consensus statement recommended that adults with gastric or duodenal ulcer disease, who are infected with H pylori, should receive antimicrobial treatment. The European Maastricht Consensus Report suggested broader indications for treatment of infected adults. It states that treatment is advisable for all H pylori infected dyspeptic patients diagnosed non-invasively under 45 years of age at a primary care level. Patients older than 45 years with dyspeptic symptoms should be treated for H pylori infection but only after endoscopy to rule out any other underlying pathology. The European guidelines also recommend treatment for infected patients with mucosa associated lymphoid tissue lymphoma and patients who are found to have intestinal metaplasia and gastric atrophy. PMID- 10457036 TI - Treatment after failure: the problem of "non-responders". AB - Although the currently most effective treatment regimens cure about 90% of infections, 10% of patients remain Helicobacter pylori positive. Several factors contribute to treatment failure. These include patient compliance, bacterial resistance to antibiotics, and treatment related issues. Treatment failure leads to the development of bacterial resistance to metronidazole and clarithromycin. Retreatment can be undertaken after considering several different strategies: to repeat the same regimen with full doses of medications and a longer treatment duration, or to choose different regimens to avoid the antibiotic previously used, or to switch to proton pump inhibitor (PPI) based quadruple therapy or ranitidine bismuth citrate (RBC) based triple therapy. In principle, full doses and longer treatment durations are advisable. As retreatment is always difficult, choosing the best available first line treatment regimen is still the best "rescue" treatment. PMID- 10457037 TI - Helicobacter pylori-reflections for the next millennium. PMID- 10457038 TI - The functional gastrointestinal disorders and the Rome II process. PMID- 10457039 TI - Fundamentals of neurogastroenterology. AB - Current concepts and basic principles of neurogastroenterology in relation to functional gastrointestinal disorders are reviewed. Neurogastroenterology is emphasized as a new and advancing subspecialty of clinical gastroenterology and digestive science. As such, it embraces the investigative sciences dealing with functions, malfunctions, and malformations in the brain and spinal cord, and the sympathetic, parasympathetic and enteric divisions of the autonomic innervation of the digestive tract. Somatomotor systems are included insofar as pharyngeal phases of swallowing and pelvic floor involvement in defecation, continence, and pelvic pain are concerned. Inclusion of basic physiology of smooth muscle, mucosal epithelium, and the enteric immune system in the neurogastroenterologic domain relates to requirements for compatibility with neural control mechanisms. Psychologic and psychiatric relations to functional gastrointestinal disorders are included because they are significant components of neurogastroenterology, especially in relation to projections of discomfort and pain to the digestive tract. PMID- 10457041 TI - Psychosocial aspects of the functional gastrointestinal disorders. AB - The functional gastrointestinal disorders (FGID) are the most frequent conditions seen in gastroenterology practice and comprise a major portion of primary care. Psychosocial factors are important in these disorders with regard to: (1) their effects on gut physiology; (2) their modulation of the symptom experience; (3) their influence on illness behavior; (4) their impact on outcome; and (5) the choice of the therapeutic approach. This paper provides a review and consensus of the existing literature by gastroenterologists, psychiatrists, psychologists, physiologists, and health services investigators. Evidence is provided to support the biopsychosocial model as a basis for understanding and treating these disorders, and epidemiological and clinical information on the relations of psychosocial factors to gut physiology, symptom presentation, health behavior, and outcome is offered. Features of motility, personality, abuse history, health concerns, and treatment-seeking differ between patients with FGID and healthy controls, but they are not specific to FGID. They occur in other patients with chronic medical conditions and/or psychiatric disorders. Review of treatment trials indicates clear support for psychotherapeutic treatments, especially in the long term, as well as some evidence for the benefit of antidepressants in FGID, even in the absence of improvements in mood. PMID- 10457040 TI - Principles of applied neurogastroenterology: physiology/motility-sensation. AB - Many of the symptoms characteristic of the functional gastrointestinal disorders (FGID) are consistent with dysfunction of the motor and/or sensory apparatus of the digestive tract. Those aspects of sensorimotor dysfunction most relevant to the FGID include alterations in: gut contractile activity; myoelectrical activity; tone and compliance; and transit, as well as an enhanced sensitivity to distension, in each region of the gastrointestinal tract. Assessment of these phenomena involves a number of techniques, some well established and others requiring further validation. Using such techniques, researchers have reported a wide range of alterations in sensory and in motor function in the FGID. Importantly, however, relationships between such dysfunction and symptoms have been relatively weak, and so the clinical relevance of the former remains unclear. Moreover, the proportions of patients in the various symptom subgroups who display dysfunction, and the extent and severity of their symptoms, require better characterization. On a positive note, progress is occurring on several fronts, especially in relation to functional dyspepsia and irritable bowel syndrome, and based on the data gathered to date, a number of areas where further advances are required can be highlighted. PMID- 10457042 TI - Functional esophageal disorders. AB - The functional esophageal disorders include globus, rumination syndrome, and symptoms that typify esophageal diseases (chest pain, heartburn, and dysphagia). Factors responsible for symptom production are poorly understood. The criteria for diagnosis rest not only on compatible symptoms but also on exclusion of structural and metabolic disorders that might mimic the functional disorders. Additionally, a functional diagnosis is precluded by the presence of a pathology based motor disorder or pathological reflux, defined by evidence of reflux esophagitis or abnormal acid exposure time during ambulatory esophageal pH monitoring. Management is largely empirical, although efficacy of psychopharmacological agents and psychological or behavioral approaches has been established for several of the functional esophageal disorders. As gastroesophageal reflux disease overlaps in presentation with most of these disorders and because symptoms are at least partially provoked by acid reflux events in many patients, antireflux therapy also plays an important role both in diagnosis and management. Further understanding of the fundamental mechanisms responsible for symptoms is a priority for future research efforts, as is the consideration of treatment outcome in a broader sense than reduction in esophageal symptoms alone. Likewise, the value of inclusive rather than restrictive diagnostic criteria that encompass other gastrointestinal and non gastrointestinal symptoms should be examined to improve the accuracy of symptom based criteria and reduce the dependence on objective testing. PMID- 10457043 TI - Functional gastroduodenal disorders. AB - While widely used in research, the 1991 Rome criteria for the gastroduodenal disorders, especially symptom subgroups in dyspepsia, remain contentious. After a comprehensive literature search, a consensus-based approach was applied, supplemented by input from international experts who reviewed the report. Three functional gastroduodenal disorders are defined. Functional dyspepsia is persistent or recurrent pain or discomfort centered in the upper abdomen; evidence of organic disease likely to explain the symptoms is absent, including at upper endoscopy. Discomfort refers to a subjective, negative feeling that may be characterized by or associated with a number of non-painful symptoms including upper abdominal fullness, early satiety, bloating, or nausea. A dyspepsia subgroup classification is proposed for research purposes, based on the predominant (most bothersome) symptom: (a) ulcer-like dyspepsia when pain (from mild to severe) is the predominant symptom, and (b) dysmotility-like dyspepsia when discomfort (not pain) is the predominant symptom. This classification is supported by recent evidence suggesting that predominant symptoms, but not symptom clusters, identify subgroups with distinct underlying pathophysiological disturbances and responses to treatment. Aerophagia is an unusual complaint characterized by air swallowing that is objectively observed and troublesome repetitive belching. Functional vomiting refers to frequent episodes of recurrent vomiting that is not self-induced nor medication induced, and occurs in the absence of eating disorders, major psychiatric diseases, abnormalities in the gut or central nervous system, or metabolic diseases that can explain the symptom. The current classification requires careful validation but the criteria should be of value in future research. PMID- 10457044 TI - Functional bowel disorders and functional abdominal pain. AB - The Rome diagnostic criteria for the functional bowel disorders and functional abdominal pain are used widely in research and practice. A committee consensus approach, including criticism from multinational expert reviewers, was used to revise the diagnostic criteria and update diagnosis and treatment recommendations, based on research results. The terminology was clarified and the diagnostic criteria and management recommendations were revised. A functional bowel disorder (FBD) is diagnosed by characteristic symptoms for at least 12 weeks during the preceding 12 months in the absence of a structural or biochemical explanation. The irritable bowel syndrome, functional abdominal bloating, functional constipation, and functional diarrhea are distinguished by symptom-based diagnostic criteria. Unspecified FBD lacks criteria for the other FBDs. Diagnostic testing is individualized, depending on patient age, primary symptom characteristics, and other clinical and laboratory features. Functional abdominal pain (FAP) is defined as either the FAP syndrome, which requires at least six months of pain with poor relation to gut function and loss of daily activities, or unspecified FAP, which lacks criteria for the FAP syndrome. An organic cause for the pain must be excluded, but aspects of the patient's pain behavior are of primary importance. Treatment of the FBDs relies upon confident diagnosis, explanation, and reassurance. Diet alteration, drug treatment, and psychotherapy may be beneficial, depending on the symptoms and psychological features. PMID- 10457045 TI - Functional disorders of the biliary tract and pancreas. AB - The term "dysfunction" defines the motor disorders of the gall bladder and the sphincter of Oddi (SO) without note of the potential etiologic factors for the difficulty to differentiate purely functional alterations from subtle structural changes. Dysfunction of the gall bladder and/or SO produces similar patterns of biliopancreatic pain and SO dysfunction may occur in the presence of the gall bladder. The symptom-based diagnostic criteria of gall bladder and SO dysfunction are episodes of severe steady pain located in the epigastrium and right upper abdominal quadrant which last at least 30 minutes. Gall bladder and SO dysfunctions can cause significant clinical symptoms but do not explain many instances of biliopancreatic type of pain. The syndrome of functional abdominal pain should be differentiated from gall bladder and SO dysfunction. In the diagnostic workup, invasive investigations should be performed only in the presence of compelling clinical evidence and after non-invasive testing has yielded negative findings. Gall bladder dysfunction is suspected when laboratory, ultrasonographic, and microscopic bile examination have excluded the presence of gallstones and other structural abnormalities. The finding of decreased gall bladder emptying at cholecystokinin-cholescintigraphy is the only objective characteristic of gall bladder dysfunction. Symptomatic manifestation of SO dysfunction may be accompanied by features of biliary obstruction (biliary-type SO dysfunction) or significant elevation of pancreatic enzymes and pancreatitis (pancreatic-type SO dysfunction). Biliary-type SO dysfunction occurs more frequently in postcholecystectomy patients who are categorized into three types. Types I and II, but not type III, have biochemical and cholangiographic features of biliary obstruction. Pancreatic-type SO dysfunction is less well classified into types. When non-invasive investigations and endoscopic retrograde cholangiopanreatography show no structural abnormality, manometry of both biliary and pancreatic sphincter may be considered. PMID- 10457046 TI - Functional disorders of the anus and rectum. AB - In this report the functional anorectal disorders, the etiology of which is currently unknown or related to the abnormal functioning of normally innervated and structurally intact muscles, are discussed. These disorders include functional fecal incontinence, functional anorectal pain, including levator ani syndrome and proctalgia fugax, and pelvic floor dyssynergia. The epidemiology of each disorder is defined and discussed, their pathophysiology is summarized and diagnostic approaches and treatment are suggested. Some suggestions for the direction of future research on these disorders are also given. PMID- 10457047 TI - Childhood functional gastrointestinal disorders. AB - This is the first attempt at defining criteria for functional gastrointestinal disorders (FGIDs) in infancy, childhood, and adolescence. The decision-making process was as for adults and consisted of arriving at consensus, based on clinical experience. This paper is intended to be a quick reference. The classification system selected differs from the one used in the adult population in that it is organized according to main complaints instead of being organ targeted. Because the child is still developing, some disorders such as toddler's diarrhea (or functional diarrhea) are linked to certain physiologic stages; others may result from behavioral responses to sphincter function acquisition such as fecal retention; others will only be recognizable after the child is cognitively mature enough to report the symptoms (e.g., dyspepsia). Infant regurgitation, rumination, and cyclic vomiting constitute the vomiting disorders. Abdominal pain disorders are classified as: functional dyspepsia, irritable bowel syndrome (IBS), functional abdominal pain, abdominal migraine, and aerophagia. Disorders of defecation include: infant dyschezia, functional constipation, functional fecal retention, and functional non-retentive fecal soiling. Some disorders, such as IBS and dyspepsia and functional abdominal pain, are exact replications of the adult criteria because there are enough data to confirm that they represent specific and similar disorders in pediatrics. Other disorders not included in the pediatric classification, such as functional biliary disorders, do occur in children; however, existing data are insufficient to warrant including them at the present time. For these disorders, it is suggested that, for the time being, clinicians refer to the criteria established for the adult population. PMID- 10457049 TI - Definition of a responder in clinical trials for functional gastrointestinal disorders: report on a symposium. PMID- 10457048 TI - Design of treatment trials for functional gastrointestinal disorders. AB - Until recently many clinical trials of functional gastrointestinal disorders (FGIDs) suffered from important weaknesses in trial design, study execution, and data analysis. This makes it difficult to determine whether truly efficacious therapies exist for these disorders. One of the important methodologic problems is the absence of validated outcome measures and lack of consensus among stakeholders on how to measure outcome. Currently much of the effort is being put into the development of validated outcome measures for several of the FGIDs. The randomized, controlled trial with parallel groups is the design of choice. In this report, guidelines are given for the basic architecture of intervention studies of FGIDs. Further studies on design issues are required to ensure the recommendations will become evidence based in the future. PMID- 10457050 TI - The road to Rome. PMID- 10457051 TI - Inactivation determinant in the I-II loop of the Ca2+ channel alpha1-subunit and beta-subunit interaction affect sensitivity for the phenylalkylamine ( )gallopamil. AB - 1. The role of calcium (Ca2+) channel inactivation in the molecular mechanism of channel block by phenylalkylamines (PAAs) was analysed in a PAA-sensitive rabbit brain class A Ca2+ channel mutant (alpha1A-PAA). Use-dependent barium current (IBa) inhibition of alpha1A-PAA by (-)gallopamil and Ca2+ channel recovery from inactivation and block were studied with two-microlectrode voltage clamp after expression of alpha1A-PAA and auxiliary alpha2-delta- and beta1a- or beta2a subunits in Xenopus oocytes. 2. Mutation Arg387Glu (alpha1A numbering) in the intracellular loop connecting domains I and II of alpha1A-PAA slowed the inactivation kinetics and reduced use-dependent inhibition (100 ms test pulses at 0.2 Hz from -80 to 20 mV) of the resulting mutant alpha1A-PAA/R-E/beta1a channels by 100 microM (-)gallopamil (53 +/- 2 %, alpha1A-PAA/beta1a vs. 31 +/- 2 %, alpha1A-PAA/R-E/beta1a, n >= 4). This amino acid substitution simultaneously accelerated the recovery of channels from inactivation and from block by ( )gallopamil. 3. Coexpression of alpha1A-PAA with the beta2a-subunit reduced fast IBa inactivation and induced a substantial reduction in use-dependent IBa inhibition by (-)gallopamil (25 +/- 4 %, alpha1A-PAA/beta2a; 13 +/- 1 %, alpha1A PAA/R-E/beta2a). The time constant of recovery from block at rest was not significantly affected. 4. These results demonstrate that changes in channel inactivation induced by Arg387Glu or beta2a-alpha1-subunit interaction affect the drug-channel interaction. PMID- 10457052 TI - Cloning and functional expression of a novel degenerin-like Na+ channel gene in mammals. AB - 1. A degenerate polymerase chain reaction (PCR) homology screening procedure was applied to rat brain cDNA in order to identify novel genes belonging to the amiloride-sensitive Na+ channel and degenerin (NaC/DEG) family of ion channels. A single gene was identified that encodes a protein related to but clearly different from the already cloned members of the family (18-30 % amino acid sequence identity). Phylogenetic analysis linked this protein to the group of ligand-gated channels that includes the mammalian acid-sensing ion channels and the Phe-Met-Arg-Phe-amide (FMRFamide)-activated Na+ channel. 2. Expression of gain-of-function mutants after cRNA injection into Xenopus laevis oocytes or transient transfection of COS cells induced large constitutive currents. The activated channel was amiloride sensitive (IC50, 1.31 microM) and displayed a low conductance (9-10 pS) and a high selectivity for Na+ over K+ (ratio of the respective permeabilities, PNa+/PK+ >= 10), all of which are characteristic of NaC/DEG channel behaviour. 3. Northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed a predominant expression of its mRNA in the small intestine, the liver (including hepatocytes) and the brain. This channel has been called the brain-liver-intestine amiloride-sensitive Na+ channel (BLINaC). 4. Corresponding gain-of-function mutations in Caenorhabditis elegans degenerins are responsible for inherited neurodegeneration in the nematode. Besides the BLINaC physiological function that remains to be established, mutations in this novel mammalian degenerin-like channel might be of pathophysiological importance in inherited neurodegeneration and liver or intestinal pathologies. PMID- 10457053 TI - Kinetics of cell lysis, dye uptake and permeability changes in cells expressing the rat P2X7 receptor. AB - 1. Extracellular ATP acting on P2X7 receptors opens a channel permeable to small cations, creates an access pathway for the entry of larger molecular weight dyes, and causes cell death. We used whole-cell recording and fluorescence microscopy to measure the time courses of ionic currents, uptake of the propidium dye YO-PRO 1, and membrane disruption, in human embryonic kidney (HEK293) cells expressing the rat P2X7 receptor. 2. The ATP analogue 2', 3'-O-(benzoyl-4-benzoyl)-ATP (30 microM) induced membrane blebbing within 30-40 s of sustained application; this was 5-10 times slower when extracellular sodium was replaced by larger cations. 3. Fluorescence of YO-PRO-1 was detectable within 3 s, and the uptake reached a steady rate within 10-20 s; YO-PRO-1 uptake was greatly enhanced by removing extracellular sodium. 4. Electrophysiological measurements of current reversal potentials with intracellular sodium and extracellular cations of different sizes showed that the ionic channel progressively t'2+LE0i%-i"dilated during 10-20 s to a diameter greater than 1 nm (10 A). With short agonist applications (3-5 s) the pore dilatation and YO-PRO-1 uptake were reversible and repeatable. 5. Polyethylene glycols having molecular weights >= 5000 blocked the increase in cation permeability, YO-PRO-1 uptake and membrane blebbing. 6. We conclude that maximum P2X7 receptor activation causes an exponential dilatation of the ion channel with a time constant of 25 s to a final diameter of 3-5 nm from an initial minimum pore diameter of 0.8 nm. PMID- 10457054 TI - ATP-sensitive K+ channel openers prevent Ca2+ overload in rat cardiac mitochondria. AB - 1. Mitochondrial dysfunction, secondary to excessive accumulation of Ca2+, has been implicated in cardiac injury. We here examined the action of potassium channel openers on mitochondrial Ca2+ homeostasis, as these cardioprotective ion channel modulators have recently been shown to target a mitochondrial ATP sensitive K+ channel. 2. In isolated cardiac mitochondria, diazoxide and pinacidil decreased the rate and magnitude of Ca2+ uptake into the mitochondrial matrix with an IC50 of 65 and 128 microM, respectively. At all stages of Ca2+ uptake, the potassium channel openers depolarized the mitochondrial membrane thereby reducing Ca2+ influx through the potential-dependent mitochondrial uniporter. 3. Diazoxide and pinacidil, in a concentration-dependent manner, also activated release of Ca2+ from mitochondria. This was prevented by cyclosporin A, an inhibitor of Ca2+ release through the mitochondrial permeability transition pore. 4. Replacement of extramitochondrial K+ with mannitol abolished the effects of diazoxide and pinacidil on mitochondrial Ca2+, while the K+ ionophore valinomycin mimicked the effects of the potassium channel openers. 5. ATP and ADP, which block K+ flux through mitochondrial ATP-sensitive K+ channels, inhibited the effects of potassium channel openers, without preventing the action of valinomycin. 6. In intact cardiomyocytes, diazoxide also induced mitochondrial depolarization and decreased mitochondrial Ca2+ content. These effects were inhibited by the mitochondrial ATP-sensitive K+ channel blocker 5-hydroxydecanoic acid. 7. Thus, potassium channel openers prevent mitochondrial Ca2+ overload by reducing the driving force for Ca2+ uptake and by activating cyclosporin sensitive Ca2+ release. In this regard, modulators of an ATP-sensitive mitochondrial K+ conductance may contribute to the maintenance of mitochondrial Ca2+ homeostasis. PMID- 10457055 TI - Substance P and bradykinin activate different types of KCa currents to hyperpolarize cultured porcine coronary artery endothelial cells. AB - 1. Substance P and bradykinin, endothelium-dependent vasodilators of pig coronary artery, trigger in endothelial cells a rise in cytosolic Ca2+ concentration ([Ca2+]i) and membrane hyperpolarization. The aim of the present study was to determine the type of Ca2+-dependent K+ (KCa) currents underlying the endothelial cell hyperpolarization. 2. The substance P-induced increase in [Ca2+]i was 30 % smaller than that induced by bradykinin, although the two peptides triggered a membrane hyperpolarization of the same amplitude. The two agonists evoked a large outward K+ current of the same conductance at maximal stimulation. Agonists applied together produced the same maximal current amplitude as either one applied alone. 3. Iberiotoxin (50 nM) reduced by about 40 % the K+ current activated by bradykinin without modifying the substance P response. Conversely, apamin (1 microM) inhibited the substance P-induced K+ current by about 65 %, without affecting the bradykinin response. Similar results were obtained on peptide-induced membrane hyperpolarization. 4. Bradykinin-induced, but not substance P-induced, endothelium-dependent relaxation resistant to NG-nitro-L arginine and indomethacin was partly inhibited by 3 microM 17-octadecynoic acid (17-ODYA), an inhibitor of cytochrome P450 epoxygenase. Similarly, the bradykinin induced K+ current was reduced by 17-ODYA. 5. Our results show that responses to substance P and bradykinin result in a hyperpolarization due to activation of different KCa currents. A current consistent with the activation of large conductance (BKCa) channels was activated only by bradykinin, whereas a current consistent with the activation of small conductance (SKCa) channels was stimulated only by substance P. The observation that a similar electrical response is produced by different pools of channels implies distinct intracellular pathways leading to KCa current activation. PMID- 10457056 TI - Tyrosine kinases modulate K+ channel gating in mouse Schwann cells. AB - 1. The whole-cell configuration of the patch-clamp technique and immunoprecipitation experiments were used to investigate the effects of tyrosine kinases on voltage-dependent K+ channel gating in cultured mouse Schwann cells. 2. Genistein, a broad-spectrum tyrosine kinase inhibitor, markedly reduced the amplitude of a slowly inactivating delayed-rectifier current (IK) and, to a lesser extent, that of a transient K+ current (IA). Similar results were obtained on IK with another tyrosine kinase inhibitor, herbimycin A. Daidzein, the inactive analogue of genistein, was without effect. 3. Unlike herbimycin A, genistein produced additional effects on IA by profoundly affecting its gating properties. These changes consisted of slower activation kinetics with an increased time to peak, a positive shift in the voltage dependence of activation (by +30 mV), a decrease in the steepness of activation gating (9 mV per e-fold change) and an acceleration of channel deactivation. 4. The steepness of the steady-state inactivation was increased by genistein treatment, while the recovery from inactivation was not significantly altered. 5. The action of genistein on voltage-dependent K+ (Kv) currents was accompanied by a decrease in tyrosine phosphorylation of Kv1.4 as well as Kv1.5 and Kv2.1 encoding transient and slowly inactivating delayed-rectifier K+ channel alpha subunits, respectively. 6. In conclusion, the present study shows that tyrosine kinases markedly affect the amplitude of voltage-dependent K+ currents in Schwann cells and finely tune the gating properties of the transient K+ current component IA. These modulations may be functionally relevant in the control of K+ channel activity during Schwann cell development and peripheral myelinogenesis. PMID- 10457058 TI - Activation by bitter substances of a cationic channel in membrane patches excised from the bullfrog taste receptor cell. AB - 1. The response to bitter-tasting substances was recorded in outside-out membrane patches excised from the taste receptor cell of the bullfrog fungiform papilla. 2. Application of a bitter-tasting substance, quinine or denatonium, induced channel openings under conditions in which none of the second messenger candidates or their precursors (e.g. cyclic nucleotide, inositol 1,4,5 trisphosphate, Ca2+, ATP and GTP) were present on either side of the membrane. The response could be recorded > 10 min after excision of the patch membrane. These data suggest that the channel was directly gated by the bitter-tasting substances. 3. No change in response was detected upon addition to the cytoplasmic side of either GDPbetaS (1 mM) or GTPgammaS (1 mM), suggesting that the G protein cascade has no direct relation to response generation. 4. The quinine-induced current was dose dependent. The lowest effective concentration was approximately 0.1 mM, and the saturating concentration was near 1 mM. The dose-response curve was fitted by the Hill equation with a K of 0.52 mM and a Hill coefficient of 3.8. 5. The single channel conductance measured in 120 mM NaCl solution was 10 pS. The channel was cation selective, and the ratio of the permeabilities for Na+, K+ and Cs+ (PNa : PK : PCs) was 1 : 0.48 : 0.39. The unitary conductance was dependent on the extracellular Ca2+ concentration ([Ca2+]o); 9.2 pS in a nominally Ca2+-free solution, and 4.5 pS in 1. 8 mM [Ca2+]o. 6. The dose dependence, the ion selectivity and the dependence of the unitary conductance on [Ca2+]o were almost identical to those of the quinine induced whole-cell current reported previously, indicating that the channel activity observed in the excised membrane is the basis of the whole-cell current. 7. The present observations suggest the new possibility that the cationic channel directly gated by bitter substances is involved in the bitter taste transduction mechanism. PMID- 10457059 TI - Activation of intrinsic hippocampal theta oscillations by acetylcholine in rat septo-hippocampal cocultures. AB - 1. Oscillatory electro-encephalographic activity at theta frequencies (4-15 Hz) can be recorded from the hippocampus in vivo and depends on intact septal projections. The hypothesis that these oscillations are imposed on the hippocampus by rhythmically active septal inputs was tested using dual intracellular recordings from CA1 and CA3 pyramidal cells in septo-hippocampal cocultures. 2. Septo-hippocampal cocultures displayed spontaneous oscillatory synaptic activity at theta frequencies. In CA3 cells, EPSP/IPSP sequences predominated, whereas only EPSPs were apparent in CA1 cells. Synaptic potentials in CA3 cells preceded those in CA1 cells by 5-10 ms. 3. Oscillatory synaptic activity was blocked in cocultures by the muscarinic antagonist atropine (0.1 microM), facilitated but unchanged in frequency upon application of the acetylcholinesterase inhibitor neostigmine (1 microM), and not seen in hippocampal monocultures. 4. The muscarinic agonist methacholine (5-20 nM) induced oscillatory synaptic activity at 4-15 Hz in hippocampal monocultures, which was identical to that occurring spontaneously in septo-hippocampal cocultures. 5. Synaptic theta activity was observed in cocultures of septal tissue with subdissected hippocampal slices containing area CA3 alone, but not in septo-CA1 cocultures. 6. We conclude that oscillatory synaptic activity at theta frequencies, with similar characteristics to theta activity in vivo, can be generated by the hippocampal network in response to activation of muscarinic receptors by synaptically released acetylcholine from septal afferents. Furthermore, the oscillatory activity is determined by mechanisms intrinsic to the hippocampal circuitry, particularly area CA3. Rhythmic septal input is not required. PMID- 10457057 TI - Na+ pump inhibition and non-selective cation channel activation by cyanide and anoxia in guinea-pig chromaffin cells. AB - 1. Hypoxia and metabolic inhibition with cyanide (CN) evoke catecholamine secretion in adrenal chromaffin cells through depolarization. We elucidated mechanisms for a CN- or anoxia-induced inward (depolarization) current, using the perforated patch method. 2. Bath application of Ba2+ induced a dose-dependent inhibition of a muscarine-induced current (IMUS) and part of the CN-induced current (ICN) with an IC50 (concentration responsible for 50 % inhibition) of 1.3 mM. The Ba2+-sensitive component was estimated to comprise 58 % of the total ICN. 3. The Ba2+-resistant component of ICN tended to increase with shifts of membrane potential from -40 to 40 mV and was markedly suppressed by exposure to a K+-free solution or 200 microM ouabain, indicating that the majority of the Ba2+ resistant component of ICN is due to suppression of the Na+ pump current (Ipump). 4. The non-Ipump component of ICN diminished progressively in K+-free solution. Substitution of glucose for sucrose in a K+-free CN solution further diminished the CN potency to produce the non-Ipump component. 5. The I-V relationship for the non-Ipump component of ICN had a reversal potential of -3 and -47 mV at 147 and 5.5 mM Na+, respectively, and showed an outward rectification, indicating that the non-Ipump component of ICN is due to activation of non-selective cation channels. 6. Exposure to anoxia induced a current with an amplitude comparable to that of ICN, and the anoxia-induced current apparently occluded development of ICN. The anoxia-induced current diminished by ca 60 % in the absence of K+ and reversed polarity at 5 mV under K+-free conditions. 7. It is concluded that exposure to CN and to anoxia induces suppression of the Na+ pump and activation of non-selective cation channels, probably due to an ATP decrease resulting mainly from consumption by the Na+ pump. PMID- 10457060 TI - Activity-dependent modulation of K+ currents at presynaptic terminals of mammalian central synapses. AB - 1. The activity-dependent regulation of presynaptic K+ currents at the CA3-CA1 synapse in the rat hippocampus was investigated during a train of evoked afferent action potentials. The waveforms of presynaptic compound action potentials (cAPs) and presynaptic Ca2+ transients ([Ca2+]pre,t) were measured with fluorescent voltage-sensitive and Ca2+-sensitive indicators in rat brain slices. 2. Under control conditions, presynaptic cAPs and the accompanying [Ca2+]pre,t displayed similar amplitudes for each stimulus, suggesting that there was no cumulative change of K+ and Ca2+ currents during the test train. However, when a subgroup of presynaptic K+ channels was blocked by a low concentration of 4-aminopyridine (4 AP, 40 microM), a significant facilitation of the [Ca2+]pre,t was observed. 3. This phenomenon was not due to a direct action of 4-AP on presynaptic Ca2+ channels, but to cumulative suppression of the K+ conductance as indicated by the corresponding change in waveforms of the cAP and presynaptic fibre volley. The observed facilitation was not an artifact by virtue of increased fibre recruitment, nor was it related to the accumulation of extracellular K+; rather, it was dependent on Ca2+ influx and stimulation frequency. The time course of recovery from facilitation was closely related to the decay of the intracellular Ca2+ concentration. 4. The facilitation was not blocked by a saturating concentration of 4-AP (8 mM) but was reduced during the application of the K+ channel blocker tetraethylammonium (TEA, 10 mM), implicating the involvement of TEA-sensitive K+ channels. Such activity-dependent suppression of presynaptic K+ conductance could lead to excessive transmitter release and might explain the hippocampal epileptiform activity that can be induced by application of 4-AP. PMID- 10457061 TI - Presynaptic alpha2-adrenoceptors control excitatory, but not inhibitory, transmission at rat hippocampal synapses. AB - 1. The effects of noradrenaline on neurotransmission at rat hippocampal synapses were investigated by recording autaptic currents in single neurons isolated on glial microislands. Noradrenaline reduced excitatory, but not inhibitory, autaptic currents in a pertussis toxin-sensitive manner, but the amine did not affect glutamate-evoked currents. 2. The inhibition of excitatory autaptic currents by noradrenaline was half-maximal at 0. 11 +/- 0.06 microM. The alpha2 adrenoceptor agonists UK 14 304 and clonidine were equipotent to noradrenaline in reducing these currents, whereas the alpha1-adrenoceptor agonist methoxamine and the beta-adrenoceptor agonist isoprenaline (isoproterenol) were ineffective. The reduction of excitatory autaptic currents by noradrenaline was not altered by the alpha1-adrenergic antagonist urapidil or the beta-antagonist propranolol, but reduced by the alpha2-antagonist yohimbine. The subtype-preferring antagonists rauwolscine and phentolamine (both at 0.3 microM) caused 9-fold and 36-fold rightward shifts in the concentration-response curve for the noradrenaline dependent reduction of excitatory autaptic currents, respectively. Prazosine (1 microM) did not affect this concentration-response curve. 3. Noradrenaline reduced voltage-activated Ca2+ currents in excitatory, but not in inhibitory, microisland neurons. For comparison, the GABAB agonist baclofen reduced both excitatory and inhibitory autaptic currents and diminished voltage-activated Ca2+ currents in both types of neurons. The inhibition of Ca2+ currents by noradrenaline was half-maximal at 0.17 +/- 0.05 microM, and UK 14 304 and clonidine were equipotent to noradrenaline in reducing these currents. The noradrenaline-induced reduction of Ca2+ currents was antagonized by yohimbine, but not by urapidil or propranolol; the subtype-preferring alpha2-adrenergic antagonists displayed the following rank order of activity: phentolamine > rauwolscine > prazosine. 4. Noradrenaline did not affect K+ currents and failed to alter the frequency of miniature excitatory postsynaptic currents measured in mass cultures of hippocampal neurons. 5. These results show that noradrenaline regulates transmission at glutamatergic, but not at GABAergic, hippocampal synapses via presynaptic alpha2-adrenoceptors of the alpha2A/D subtype. This inhibitory action involves an inhibition of voltage-activated Ca2+ currents, but no modulation of spontaneous vesicle exocytosis or of voltage-activated K+ currents. PMID- 10457062 TI - Glutamate-induced mitochondrial depolarisation and perturbation of calcium homeostasis in cultured rat hippocampal neurones. AB - 1. The objective of this study was to clarify the relationships between loss of mitochondrial potential and the perturbation of neuronal Ca2+ homeostasis induced by a toxic glutamate challenge. Digital fluorescence imaging techniques were employed to monitor simultaneously changes in cytoplasmic Ca2+ concentration ([Ca2+]i) and mitochondrial potential (DeltaPsim) in individual hippocampal neurones in culture coloaded with fura-2 AM or fura-2FF AM and rhodamine 123 (Rh 123). 2. In most cells (96 %) at 6-7 days in vitro (DIV) and in a small proportion of cells (29 %) at 11-17 DIV the [Ca2+]i increase induced by exposure to 100 microM glutamate for 10 min was associated with a small mitochondrial depolarisation, followed by mitochondrial repolarisation, and a degree of recovery of [Ca2+]i following glutamate washout. In the majority of neurones at 11-17 DIV (71 %), exposure to glutamate for 10 min induced a profound mono- or biphasic mitochondrial depolarisation, which was clearly correlated with a sustained [Ca2+]i plateau despite the removal of glutamate. 3. Addition of glutamate receptor antagonists (15 microM MK-801 plus 75 microM 6-cyano-7 nitroquinoxaline-2, 3-dione (CNQX)) to the washout solution did not affect the post-glutamate [Ca2+]i plateau in neurones exhibiting a profound mitochondrial depolarisation but greatly improved [Ca2+]i recovery in those neurones undergoing only a small mitochondrial depolarisation, suggesting that the release of endogenous glutamate delays [Ca2+]i recovery in the postglutamate period. 4. Cyclosporin A (500 nM) or N-methyl Val-4-cyclosporin A (200 nM) delayed or even prevented the development of the second phase of mitochondrial depolarisation in cells at 11-17 DIV and increased the proportion of neurones exhibiting a small monophasic mitochondrial depolarisation and [Ca2+]i recovery upon glutamate removal. 5. We have thus described a striking correlation between mitochondrial depolarisation and the failure of cells to restore [Ca2+]i following a toxic glutamate challenge. These data suggest that mitochondrial dysfunction plays a major role in the deregulation of [Ca2+]i associated with glutamate toxicity. PMID- 10457063 TI - Distinct temporal profiles of activity-dependent calcium increase in pyramidal neurons of the rat visual cortex. AB - 1. Using fluo-3-based fluorometry, we studied variation in depolarization-induced calcium increases in the proximal dendrites or soma of pyramidal neurons in layer II/III of the rat visual cortex. 2. Depolarization for all durations tested (0.1 2 s; 0.5 nA) evoked a train of action potentials and a small increase in calcium signal (mean 26 %) which peaked within 1 s of the onset of depolarization. With depolarization for longer than 1 s, this small increase was often followed by a larger increase (73 %). This later phase of calcium increase occurred without sudden changes in action potential firing. 3. Application of ryanodine, which suppresses intracellular calcium release, abolished the second phase without affecting the early phase in a use-dependent manner. Meanwhile, no major changes were observed in the pattern of action potential firing. 4. In calcium-free medium, both the early and late phases were almost undetectable, although action potential firing was still evoked by injection of depolarizing currents. Since the late phase depended on intracellular calcium release, this effect of calcium free medium on the late phase is likely to be indirect through an influence on the early phase. 5. This two-phase profile was observed with somatic depolarization or with antidromic action potentials induced by tetanization. Neocortical pyramidal neurons can thus recruit calcium from different sources, even without chemical sensitization, generating temporally diverse profiles of intracellular calcium signal in response to action potential firing. 6. Such variety in the mechanisms of calcium increase may be relevant to the role of calcium as a versatile second messenger for various types of synaptic plasticity. PMID- 10457064 TI - Group III metabotropic glutamate receptors control corticothalamic synaptic transmission in the rat thalamus in vitro. AB - 1. Corticothalamic (CT) EPSPs evoked at <= 0.1 Hz were recorded from thalamocortical neurones in the rat dorsal lateral geniculate nucleus in vitro, with both GABAA and GABAB receptors blocked. 2. The group III metabotropic glutamate (mGlu) receptor agonists L-2-amino-4-phosphono-butyric acid (L-AP4) and O-phospho-L-serine (L-SOP) both caused a concentration-dependent depression of the CT EPSP. The maximum depression and EC50 values for these effects were 64.4 +/- 3.8 % and 88.0 +/- 24.7 microM for L-AP4, and 42.0 +/- 2.5 % and 958 +/- 492 microM for L-SOP, respectively (means +/- s.e.m.). Neither agonist had any effect on membrane potential or input resistance. 3. The depression of the CT EPSP caused by L-AP4 was reversed using the group III antagonist (S)-2-amino-2-methyl 4-phosphonobutanoic acid (MAP4, 1 mM), and the group II/III antagonist LY341495 (3 microM), but not using the group II antagonist (2S)-alpha-ethylglutamic acid (300 microM). The potencies of L-AP4, L-SOP and LY341495 indicate that this action of L-AP4 is mediated via mGlu7 and mGlu8 and not mGlu4 receptors. 4. Neither MAP4 nor LY341495 had any effect on the CT EPSPs evoked by 10 Hz trains of five stimuli, indicating the lack of endogenous activation of group III mGlu receptors in the thalamus during short bursts of cortical input. However, the magnitude of the depression caused by L-AP4 indicates that any physiological activation of group III mGlu receptors would have a profound effect on the CT input to the thalamus, and hence cortical control of thalamic function. PMID- 10457065 TI - Modulation of the hyperpolarization-activated cation current of rat thalamic relay neurones by intracellular pH. AB - 1. Properties of the hyperpolarization-activated cation current (Ih) were investigated in thalamocortical neurones of an in vitro slice preparation of the rat ventrobasal thalamic complex (VB) before and during changes of pipette pH (pHp), intracellular pH (pHi) and bath pH (pHb) using the whole-cell patch-clamp technique and fluorescence ratio imaging of the pH indicator 2',7' bis(carboxyethyl)-5(and -6)-carboxyfluorescein (BCECF). 2. Recording of Ih with predefined pHp revealed significant shifts in the voltage dependence of Ih activation (V ) of 4-5 mV to more positive values for a pHp of 7.5 and 2-3 mV to more negative values for a pHp of 6.7 as compared to control values (pHp = 7.1). 3. Application of the weak acid lactate (20 mM), which produced a slow monophasic intracellular acidification, induced a reversible negative shift of V of up to 3 mV. Application of 20 mM TMA, which caused a distinct intracellular alkalinization, shifted V to 4-5 mV more positive values. 4. In slices bathed in Hepes-buffered saline, no significant pHo dependence of Ih was observed. Changing pHo by altering the extracellular [HCO3-] in the presence of constant pCO2 also revealed no significant pHo dependence of Ih. 5. Rhythmic stimulation of thalamocortical neurones with repetitive depolarizing pulse trains caused an intracellular acidification, which reversibly decreased the amplitude and time course of activation of Ih. 6. The results of the present study indicate that shifts in pHi result in a significant modulation of the gating properties of Ih channels in TC neurones. Through this mechanism activity-dependent shifts in pHi may contribute to the up- and downregulation of Ih. PMID- 10457066 TI - Upregulation of the hyperpolarization-activated cation current in rat thalamic relay neurones by acetazolamide. AB - 1. The effect of inhibition of brain carbonic anhydrase (CA) on the hyperpolarization-activated cation current (Ih) of thalamocortical (TC) neurones of the rat ventrobasal thalamic complex (VB) was investigated in an in vitro slice preparation using the whole-cell patch-clamp technique and fluorescence ratio imaging of the pH indicator 2',7'-bis(carboxyethyl)-5(and -6) carboxyfluorescein (BCECF). 2. Recording of Ih before and after addition of 0.4 0.8 mM acetazolamide to the bathing fluid revealed a significant shift in the voltage dependence of activation (V ) of 5-7 mV to more positive potentials. 3. Simultaneous recording of Ih and BCECF fluorescence ratio (F420/F495) revealed an increase in Ih amplitude accompanied by an intracellular alkalinization upon application of acetazolamide. The CA inhibitor ethoxyzolamide (EZA, 50 microM) also led to an intracellular alkalinization and a subsequent 4-5 mV positive shift of V of Ih. 4. Acetazolamide and EZA both profoundly slowed the rapid fall of pHi upon switching from Hepes- to CO2/HCO3--buffered superfusate, indicating intracellular CA isoforms in TC neurones. 5. In slices bathed in Hepes-buffered saline, addition of acetazolamide had no effect on the amplitude and time course of activation of Ih, indicating that the action of acetazolamide on Ih was dependent on the presence of HCO3-. 6. Under current-clamp conditions, the neuronal response to hyperpolarizing current pulses in the presence of acetazolamide was decreased as compared to control. This resulted in a strongly reduced ability of TC neurones to produce rebound Ca2+-mediated spikes. 7. The present results implied that in TC neurones acetazolamide led to an intracellular alkalinization which causes, due to its pH sensitivity, an increase in Ih. PMID- 10457067 TI - The force bearing capacity of frog muscle fibres during stretch: its relation to sarcomere length and fibre width. AB - 1. Single fibres isolated from the anterior tibialis muscle of Rana temporaria were tetanized (0.9-1.8 C) while a marked ( approximately 1 mm) segment was held at constant length by feedback control. Force enhancement was produced by applying a controlled stretch ramp to the fibre segment during the tetanus plateau, the steady force reached during stretch being used as a measure of the maximum force that the myosin cross-bridges can hold before they detach. 2. The amplitude of force enhancement during stretch did not vary in proportion to the isometric force as the sarcomere length was changed, maximum force enhancement being attained near 2.4 microm sarcomere length compared with 2.0 microm for the isometric force. 3. The influence of fibre width on the force enhancement sarcomere length relationship was evaluated by normalizing force enhancement to the tetanic (pre-stretch) force in this way allowing for the differences in myofilament overlap at the various lengths. The amplitude of force enhancement (normalized to the tetanic force) increased by approximately 70 % as the relative width of the myofilament lattice was reduced from a nominal value of 1.05 at a sarcomere length of 1.8 microm to 0.85 at a sarcomere length of 2.8 microm. 4. Changes in fibre width equivalent to those produced by altering the sarcomere length were produced by varying the tonicity of the extracellular medium. Force enhancement, normalized to the control isometric force at each tonicity, exhibited a width dependence that agreed well with that described in the previous point. Stretch ramps applied to frog skinned muscle fibres during calcium-induced contracture likewise resulted in a greater force enhancement during stretch after reducing the fibre width by osmotic compression. 5. The results suggest that the strength of binding of the myosin cross-bridges, unlike the isometric force, varies with the lateral distance between the myofilaments. PMID- 10457068 TI - F-actin stabilization increases tension cost during contraction of permeabilized airway smooth muscle in dogs. AB - 1. Dynamic actin reorganization involving actin polymerization and depolymerization may play an important functional role in smooth muscle. 2. This study tested the hypothesis that F-actin stabilization by phalloidin increases tension cost (i.e. ATP hydrolysis rate per unit of isometric force) during Ca2+ induced activation of Triton X-100-permeabilized canine tracheal smooth muscle. 3. Adenosine 5'-triphosphate (ATP) hydrolysis rate was quantified using an enzyme coupled NADH fluorometric technique, regulatory myosin light chain (rMLC) phosphorylation was measured by Western blot analysis, and maximum unloaded shortening velocity (Vmax) was estimated by interpolation of the force-velocity relationship to zero load during isotonic loading. 4. Maximal activation with 10 microM free Ca2+ induced sustained increases in isometric force, stiffness, and rMLC phosphorylation. However, the increase in ATP hydrolysis rate initially reached peak values, but then declined to steady-state levels above that of the unstimulated muscle. Thus, tension cost decreased throughout steady-state isometric force. 5. Following incubation of permeabilized strips with 50 microM phalloidin for 1 h, the increases in isometric force and stiffness were not sustained despite a sustained increase in rMLC phosphorylation. Also, after an initial decline, tension cost increased throughout activation. Phalloidin had no effect on Vmax during steady-state isometric force or on rMLC phosphorylation. 6. These findings suggest that dynamic reorganization of actin is necessary for optimal energy utilization during contraction of permeabilized airway smooth muscle. PMID- 10457069 TI - Sympathetic inhibition of ascending and descending interneurones during the peristaltic reflex in the isolated guinea-pig distal colon. AB - 1. We investigated the effects of sympathetic nerve stimulation within ascending and descending reflex pathways underlying the peristaltic reflex in the guinea pig distal colon. 2. A three-chambered partitioned bath was used to divide a segment of distal colon into stimulation, recording and intermediate regions. The effects of lumbar colonic nerves (LCN) could be localized to the intermediate region by surgical lesions of the mesentery and by application of guanethidine (3 microM) to the stimulation and recording chambers. 3. Brush stroking the mucosa in the anal and oral stimulation chambers elicited a synchronous contraction of the longitudinal muscle (LM) and circular muscle (CM) oral to, and transient relaxation of the LM and CM anal to, the stimulus, respectively. 4. After N omega nitro-L-arginine (L-NA; 100 microM) in the oral and intermediate chambers, mucosal stimulation in the oral chamber elicited a prolonged descending inhibitory and excitatory complex in both the LM and CM in the anal recording chamber. This was blocked by hexamethonium (300 microM), which did not affect the transient relaxation response recorded in control conditions. 5. Stimulation of the LCN (1200 pulses, 20 Hz), delivered to the intermediate region, abolished the oral contraction and the L-NA-induced anal complex in both the LM and CM, but was without effect on the transient hexamethonium-resistant anal relaxation. These effects of LCN stimulation were reversed by phentolamine (3 microM) or yohimbine (100 nM), but not propranolol (10 microM), when added to the intermediate chamber. 6. LCN stimuli (2-20 Hz, 600 micros pulses) directed to the recording chamber elicited synchronous relaxations in the LM and CM that were unaffected by hexamethonium (300 microM), but were reduced by yohimbine and usually blocked by the further addition of propranolol (10 microM). 7. In conclusion, sympathetic nerve stimulation inhibits orally and anally projecting cholinergic interneurones underlying the peristaltic reflex in the distal colon. In addition, the LM and CM relax synchronously following release of sympathetic neurotransmitter, over a range of stimulus frequencies. PMID- 10457071 TI - Signalling of static and dynamic features of muscle spindle input by external cuneate neurones in the cat. AB - 1. The present experiments examined the capacity of external cuneate nucleus (ECN) neurones in the anaesthetized cat to respond to static and vibrotactile stretch of forearm extensor muscles. The aim was to compare their signalling capacities with the known properties of main cuneate neurones in order to determine whether there is differential processing of muscle spindle inputs at these parallel relay sites. 2. Static stretch (<= 2 mm in amplitude) and sinusoidal vibration were applied longitudinally to individual muscle tendons and responses recorded from single ECN neurones. The muscle-related ECN neurones that were sampled displayed a high sensitivity to both static and dynamic components of stretch, including muscle vibration at frequencies of 50-800 Hz, consistent with their dominant input being derived from primary spindle afferent fibres. 3. In response to ramp-and-hold muscle stretch, ECN neurones resembled their main cuneate counterparts in the pattern of their responses and in quantitative response measures. Their coefficients of variation in interspike intervals during steady stretch ranged from approximately 0.3 to 0.7, as they do in main cuneate responses, and their stimulus-response relations were graded as a function of stretch magnitude with low variability in responses at a fixed stretch amplitude. 4. In response to muscle vibration, ECN activity was tightly phase locked to the vibration waveform, in particular at frequencies of <= 150 Hz, where vector strength measures (R) were high (R >= 0.8) before declining as a function of frequency, with R values of approximately 0.6 at 300 Hz and <= 0.4 at 800 Hz. Both the qualitative and quantitative aspects of ECN responsiveness to the vibro stretch disturbances were indistinguishable from those of the main cuneate neurones. 5. The results demonstrate a high transmission fidelity for muscle signals across the ECN and no evidence for differential synaptic transmission across the parallel main and external cuneate nuclei. Earlier limitations observed in the capacity of cerebellar Purkinje cells to respond to primary spindle inputs must therefore be imposed at synapses within the cerebellum. PMID- 10457070 TI - Luminal ATP stimulates fluid and HCO3- secretion in guinea-pig pancreatic duct. AB - 1. The location of purinoceptors in the pancreatic duct and their role in regulating ductal secretion have been investigated by applying ATP and UTP to basolateral and luminal surfaces of pancreatic ducts isolated from the guinea-pig pancreas. 2. Changes in intracellular Ca2+ concentration were measured by microfluorometry in microperfused interlobular duct segments. Fluid and HCO3- secretion were estimated by monitoring luminal pH and luminal volume in sealed duct segments microinjected with BCECF-dextran. 3. Both ATP and UTP (1 microM) caused biphasic increases in intracellular Ca2+ concentration in pancreatic duct cells when applied to either the basolateral or luminal membrane. 4. Luminal application of both ATP and UTP evoked fluid and HCO3- secretion. The maximum response to 1 microM ATP or UTP was about 75 % of that evoked by secretin. By contrast, basolateral application of ATP or UTP inhibited spontaneous secretion by 52 % and 73 %, respectively, and secretin-evoked secretion by 41 % and 38 %, respectively. 5. The data suggest that luminal nucleotides may act in an autocrine or paracrine fashion to enhance ductal secretion while basolateral nucleotides, perhaps released from nerve terminals, may have an inhibitory effect. The fact that both apical and basolateral purinoceptors elevate intracellular Ca2+, but that they have opposite effects on secretion, suggests that additional signalling pathways are involved. PMID- 10457072 TI - Repetitive hypoxia rapidly depresses cardio-respiratory responses during active sleep but not quiet sleep in the newborn lamb. AB - 1. Arousal from sleep is an important protective response to hypoxia that becomes rapidly depressed in active sleep (AS) when hypoxia is repeated. This study questioned whether there might also be selective depression of cardio-respiratory responses to hypoxia during AS. 2. Nine newborn lambs (7-22 days of age) were studied over three successive nights. The first and third nights were baseline studies (inspired oxygen fraction, Fi,O2 = 0.21). During the second night, during every epoch of sleep, lambs were exposed to 60 s episodes of isocapnic hypoxia (Fi,O2 = 0.10). 3. During quiet sleep (QS), the probability of arousal in hypoxia exceeded the probability of spontaneous arousal (P < 0.001) throughout repeated exposures to hypoxia. Similarly, there were persisting increases in ventilation (135 +/- 25 %), blood pressure (3 +/- 1 %) and heart rate (3 +/- 1 %). 4. By contrast, rapid depression of all responses occurred during repetitive hypoxia in AS. Thus, the probability of arousal in hypoxia exceeded the probability of spontaneous arousal during the first 10 hypoxia exposures (P < 0.001) but not thereafter. Similarly, during the first 10 exposures to hypoxia, the changes in ventilation (88 +/- 15 %) and blood pressure (5 +/- 1 %) were greater than subsequent responses (P < 0.05). 5. We conclude that, when repeated, hypoxia rapidly becomes ineffective in stimulating protective arousal, ventilatory and blood pressure responses in AS, but not in QS. Selective depression of responses during AS may render the newborn particularly vulnerable to hypoxia in this state. PMID- 10457073 TI - Pulmonary stretch receptor discharges and vagal regulation of respiration differ between two mouse strains. AB - 1. Experiments were performed on adult pentobarbitone-anaesthetized mice of the OF1 and the C3H/HeJ (C3H) strains, to analyse the regulation of respiration by pulmonary stretch receptors (PSRs). 2. Although the mean respiratory period, inspiratory and expiratory durations, and tidal volume did not differ significantly between the two strains, the inspiratory onset was drastically inhibited in OF1 mice but only slightly inhibited in C3H mice in response to tracheal occlusion performed at the very end of inspiration. 3. Low current electrical stimulation of the vagus nerve induced inspiratory onset inhibition in both strains, suggesting that the weak inspiratory onset inhibition elicited by tracheal occlusion in C3H mice did not originate from a low sensitivity of the respiratory centres to PSRs. 4. During normal respiration, PSR firing rate increased with tidal volume, but reached significantly higher values in OF1 than C3H mice. During tracheal occlusion, PSR firing rate was significantly higher at the end of inspiration and during the first third of the occlusion period in OF1 than C3H mice. 5. The airway pressure resistance was significantly higher in OF1 than C3H mice. After abolishing the tracheo-bronchial muscle tone with atropine in OF1 mice, tracheal occlusions induced weak inspiratory onset inhibitions resembling the C3H mouse responses. 6. The possibility that differences in tracheo-bronchial tone between OF1 and C3H mice may lead to a greater PSR discharge and thus to a powerful inhibition on the OF1 medullary respiratory centres during tracheal occlusion is discussed. PMID- 10457074 TI - Effects of CO2 and H+ on laryngeal receptor activity in the perfused larynx in anaesthetized cats. AB - 1. Intralaryngeal CO2 reflexly decreases ventilation and increases upper airway muscle activity. Topical anaesthesia of the laryngeal mucosa or cutting the superior laryngeal nerves (SLNs) abolishes these reflexes, indicating that the receptors responsible are superficially located and that their afferent fibres are in the SLN. Intralaryngeal CO2 affects the activity of receptors recorded from the SLN. 2. An isolated, luminally perfused laryngeal preparation was developed in anaesthetized, paralysed cats in order to compare the effects of solutions with varying levels of pH and PCO2 on pressure-sensitive laryngeal receptor activity. Since the pH of tracheal surface fluid is reported to be approximately 7.0, two neutral (pH 7.4 and 7.0) and two acidic (pH 6.8 and 6.3) solutions were used. 3. Compared with neutral acapnic control solutions, neutral hypercapnic (PCO2 64 mmHg) solutions either excited or inhibited the discharge of 113 out of 211 pressure-sensitive SLN afferents. In 24 receptors, the effects of hypercapnic solutions with either neutral or acidic pH were similar in both direction and magnitude. In 50 receptors affected by neutral hypercapnic solutions, acidic acapnic solutions had no effect on 66 % of units and significantly smaller effects in the remaining units. In 17 receptors, the effects of neutral solutions with a PCO2 of 35 mmHg were significantly less than for neutral solution with a PCO2 of 64 mmHg. 4. These results show that the effects of CO2 on laryngeal pressure-sensitive receptors are independent of the pH of the perfusing media, and suggest that acidification of the receptor cell or its microenvironment is the main mechanism of CO2 chemoreception. PMID- 10457075 TI - Effect of co-activation of tongue protrudor and retractor muscles on tongue movements and pharyngeal airflow mechanics in the rat. AB - 1. The purpose of these experiments was to examine the mechanisms by which either co-activation or independent activation of tongue protrudor and retractor muscles influence upper airway flow mechanics. We studied the influence of selective hypoglossal (XIIth) nerve stimulation on tongue movements and flow mechanics in anaesthetized rats that were prepared with an isolated upper airway. In this preparation, both nasal and oral flow pathways are available. 2. Inspiratory flow limitation was achieved by rapidly lowering hypopharyngeal pressure (Php) with a vacuum pump, and the maximal rate of flow (VI,max) and the nasopharyngeal pressure associated with flow limitation (Pcrit) were measured. These experimental trials were repeated while nerve branches innervating tongue protrudor (genioglossus; medial XIIth nerve branch) and retractor (hyoglossus and styloglossus; lateral XIIth nerve branch) muscles were stimulated either simultaneously or independently at frequencies ranging from 20-100 Hz. Co activating the protrudor and retractor muscles produced tongue retraction, whereas independently activating the genioglossus resulted in tongue protrusion. 3. Co-activation of tongue protrudor and retractor muscles increased VI, max (peak increase 44 %, P < 0.05), made Pcrit more negative (peak decrease of 44 %, P < 0.05), and did not change upstream nasopharyngeal resistance (Rn). Independent protrudor muscle stimulation increased VI,max (peak increase 61 %, P < 0.05), did not change Pcrit, and decreased Rn (peak decrease of 41 %, P < 0.05). Independent retractor muscle stimulation did not significantly alter flow mechanics. Changes in Pcrit and VI,max at all stimulation frequencies were significantly correlated during co-activation of protrudor and retractor muscles (r2 = 0.63, P < 0.05), but not during independent protrudor muscle stimulation (r2 = 0.09). 4. These findings indicate that either co-activation of protrudor and retractor muscles or independent activation of protrudor muscles can improve upper airway flow mechanics, although the underlying mechanisms are different. We suggest that co-activation decreases pharyngeal collapsibility but does not dilate the pharyngeal airway. In contrast, unopposed tongue protrusion dilates the oropharynx, but has a minimal effect on pharyngeal airway collapsibility. PMID- 10457076 TI - Contractile activation characteristics of single permeabilized fibres from levator palpebrae superioris, orbicularis oculi and vastus lateralis muscles from humans. AB - 1. We investigated the contractile activation characteristics of single membrane permeabilized fibres from the following muscles from humans: the levator palpebrae superioris (LPS), an extraocular muscle; the orbicularis oculi (OO), a facial muscle; and the vastus lateralis (VL), a major muscle of the thigh. 2. Single permeabilized muscle fibres were isolated from each of the different muscles, attached to a sensitive force transducer and activated by rapid immersion in buffered solutions of varying [Ca2+] and [Sr2+]. Fibres were allocated into discrete populations based on their contractile characteristics, including their differential force responses during Ca2+ and Sr2+ activation. 3. With the exception of one fibre from the LPS, all 152 fibres sampled from the three different human muscles could be classified into either population I (slow, type I) or population II (fast, type II) based on their force-pCa(pSr) relations. The LPS muscle fibre which was unable to be classified into the two major fibre populations displayed a combination of the typical force-pCa(pSr) relations for mammalian fast and slow muscle fibres. 4. Although fibres from the LPS, OO and VL muscles had similar differential sensitivities to Ca2+and Sr2+, the steepness of the force-pCa(pSr) curves for fibres from the LPS and OO muscles were highly variable compared with those for fibres from the VL muscle. Specific forces (N cm 2) of the smaller diameter fibres from the LPS and OO muscles were significantly lower than those of fibres from the VL muscle. 5. The differences in the contractile activation characteristics between fibres from the VL muscle and those of fibres from facial (OO) muscles and extraocular (LPS) muscles, reflect the differences in their fibre composition that are responsible for their functional specificity. PMID- 10457077 TI - Contribution of the leg vasculature to hypotensive effects of an antiorthostatic posture change in humans. AB - 1. Previous results from our laboratory have shown that vasodilatation in the legs prevents mean arterial pressure (MAP) from increasing during water immersion. Therefore, we tested the hypothesis that vasodilatation in the legs is necessary for the hypotensive effects to occur during a moderate antiorthostatic posture change. 2. Ten healthy males underwent a 5 min posture change from upright seated to horizontal supine (SUP) and back to seated again with (OCCL SUP) and without simultaneous total arterial (154 +/- 1 mmHg) thigh occlusion, and a control seated period, also with and without arterial occlusion. Cardiac output (CO) was measured by a non-invasive foreign (N2O) gas rebreathing technique. 3. MAP (brachial auscultation) decreased during SUP from 94 +/- 3 to 84 +/- 2 mmHg (P < 0.0001) and total peripheral vascular resistance (TPR = MAP/CO, n = 8) decreased by 15 +/- 4 % (P < 0.001). During OCCL-SUP, MAP decreased from 98 +/- 2 to 90 +/- 2 mmHg (P < 0.005) and TPR decreased by 14 +/- 3 % (P < 0.01). 4. In conclusion, vasodilatation in the legs is not necessary for the decrease in MAP to occur during a moderate antiorthostatic manoeuvre. Therefore, vasodilatation in more central vascular beds (e.g. abdomen) can alone account for the hypotensive effects. PMID- 10457078 TI - Intracellular spreading of second messengers. PMID- 10457079 TI - Biophysical properties of mouse connexin30 gap junction channels studied in transfected human HeLa cells. AB - 1. Human HeLa cells expressing mouse connexin30 (Cx30) were used to study the electrical properties of Cx30 gap junction channels. Experiments were performed on cell pairs with the dual voltage-clamp method. 2. The gap junction conductance (gj) at steady state showed a bell-shaped dependence on junctional voltage (Vj; Boltzmann fit: Vj,0 = 27 mV, gj,min = 0.15, z = 4). The instantaneous gj decreased slightly with increasing Vj. 3. The gap junction currents (Ij) declined with time following a single exponential. The time constants of Ij inactivation (taui) decreased with increasing Vj. 4. Single channels exhibited a main state, a residual state and a closed state. The conductances gammaj,main and gammaj,residual were 179 and 48 pS, respectively (pipette solution, potassium aspartate; temperature, 36-37 degrees C; extrapolated to Vj = 0 mV). 5. The conductances gammaj,residual and gammaj,main showed a slight Vj dependence and were sensitive to temperature (Q10 values of 1.28 and 1.16, respectively). 6. Current transitions between open states (i.e. main state, substates, residual state) were fast (< 2 ms), while those between an open state and the closed state were slow (12 ms). 7. The open channel probability (Po) at steady state decreased from 1 to 0 with increasing Vj (Boltzmann fit: Vj,0 = 37 mV; z = 3). 8. Histograms of channel open times implied the presence of a single main state; histograms of channel closed times suggested the existence of two closed states (i.e. residual states). 9. We conclude that Cx30 channels are controlled by two types of gates, a fast one responsible for Vj gating involving transitions between open states (i.e. residual state, main state), and a slow one correlated with chemical gating involving transitions between the closed state and an open state. PMID- 10457080 TI - A novel cGMP-regulated K+ channel in immortalized human kidney epitheliall cells (IHKE-1). AB - 1. K+ channels from the apical membrane of immortalized human kidney epithelial (IHKE-1) cells were investigated in the cell-attached membrane configuration as well as in excised membranes using the patch clamp technique. 2. In cell-attached membrane patches the open probability (Po) of the K+ channel was 0.42 +/- 0.06 (mean +/- s.e.m. , n = 22) and its conductance was 94 +/- 5 pS with 145 mM K+ in the pipette (n = 25). In excised membrane patches the Po of the channel was 0.55 +/- 0.03 (n = 86) and its conductance was 65 +/- 2 pS (n = 68) with 145 mM K+ on one side of the membrane and 3.6 mM K+ on the other. The I-V curve of the K+ channel was not rectifying. 3. The channel was inhibited by several blockers of K+ channels such as 1 mM Ba2+ (cell-attached membrane: 78 +/- 8 %, n = 9; excised: 80 +/- 4 %, n = 26), 10 mM TEA+ (excised inside-out: 48 +/- 5 %, n = 34; excised outside-out: 100 +/- 0 %, n = 26), 0.1 mM verapamil (excised: 73 +/- 9 %, n = 12), and 10 nM charybdotoxin (excised outside-out: 67 +/- 9 %, n = 9). 4. The K+ channel was activated by depolarization and rising cytosolic Ca2+. Half maximal activity occurred at a cytosolic Ca2+ concentration of 200 nM. In the cell-attached membrane configuration the K+ channel was inhibited in a concentration-dependent manner by atrial natriuretic peptide (ANP). Powas blocked equally well by 10 nM ANP (52 +/- 7 %, n = 10), brain natriuretic peptide (BNP; 37 +/- 11 %, n = 6) and C-type natriuretic peptide (CNP; 44 +/- 13 %, n = 8). 8 Bromoguanosine 3',5' cyclic monophosphate (8-Br-cGMP, 0.1 mM) also inhibited Poof this K+ channel, by 70 +/- 10 % (n = 5). 5. In excised membrane patches cGMP inhibited Po of this K+ channel in a concentration-dependent manner. The first significant effects were measured at a concentration of 1 microM (22 +/- 7 %, n = 6), and greatest effects were obtained at 0.1 mM (34 +/- 5 %, n = 15). cAMP (0.1 mM, n = 5) as well as GTP (0.1 mM, n = 5) had no significant effects on Po of this K+ channel. ATP (0.1 mM) had a weak inhibitory effect (17 +/- 5 %, n = 14). Addition of Mg-ATP to cGMP did not increase the inhibitory effect (30 +/- 4 %, n = 14). KT5823 (1 microM), a specific inhibitor of cGMP-dependent protein kinases, did not significantly alter the cGMP-induced reduction in Po of the K+ channel in three excised membrane patches. 6. The results present the first electrophysiological characterization of a mammalian K+ channel that is directly regulated by cGMP. PMID- 10457081 TI - Serotonin via 5-HT1B and 5-HT2B receptors stimulates anion secretion in the rat epididymal epithelium. AB - 1. The short-circuit current (Isc) technique was used to study the role of 5 hydroxytryptamine (5-HT) in the regulation of anion secretion in cultured rat cauda epididymal epithelia. 2. 5-HT, the 5-HT1B-selective agonist 5 nonyloxytryptamine (5-NOT) and the 5-HT2B-selective agonist alpha-methyl-5 hydroxytryptamine (alpha-methyl-5-HT) added basolaterally stimulated Isc in a dose-dependent manner with EC50 values of 0.4, 20 and 0.3 microM, respectively. No other agonists for 5-HT receptors had any effect. 3. The pattern of responses to 5-HT was biphasic. Pretreating the tissues with the 5-HT1B-selective antagonist isamoltane (200 microM) and the 5-HT2B-selective antagonist rauwolscine (200 microM) inhibited the rapid transient phase by 55 and 45 %, whereas the sustained phase could only be blocked by rauwolscine. 4. Removal of chloride or bicarbonate or both from the normal Krebs-Henseleit solution reduced the responses to 5-HT, 5-NOT and alpha-methyl-5-HT to varying degrees. The results suggest that 5-HT1B- and 5-HT2B-mediated responses were mainly due to chloride and bicarbonate secretion, respectively. 5. Manipulation of the cAMP and Ca2+ signal transduction pathways with chemical agents provided evidence that the responses to 5-HT were mediated through cAMP. 6. Piroxicam pretreatment abolished the Isc response to alpha-methyl-5-HT but not to 5-NOT, indicating that the 5 HT2B-mediated response, but not the 5-HT1B-mediated response, is dependent on prostaglandin synthesis. 7. Immunohistochemical studies showed that 5-HT-like immunoreactivity was detected in nerve fibres and in small granular cells surrounding the epididymal tubules. 8. It is suggested that the 5-HT released from serotonergic nerve endings and/or from mast cells regulates electrolyte and fluid secretion in the epididymis. PMID- 10457083 TI - Longitudinal spread of second messenger signals in isolated rod outer segments of lizards. AB - 1. In vertebrate rods activation of the phototransduction cascade by light triggers changes in the concentrations of at least two diffusible intracellular second messengers (cGMP and Ca2+) whose actions depend on how far they spread from their site of production or entry. To address questions about their spatial spread, cell-attached patch current recording and fluorescence imaging of Calcium Green-dextran were used to measure the longitudinal spread of cGMP and Ca2+, respectively, in functionally intact isolated Gecko gecko lizard rod outer segments under whole-cell voltage clamp. 2. The light-evoked changes in cGMP and Ca2+ concentrations decayed with distance from a site of steady focal activation by two-photon absorption of 1064 nm light with similar decay lengths of approximately 3.5 microm. 3. These results can be understood on the basis of a quantitative model of coupled diffusible intracellular messengers, which is likely to have broad relevance for second messenger signalling pathways in general. 4. The decay length for the spread of adaptation from a site of steady local illumination was about 8 microm, i.e. substantially longer than the decay lengths measured for the spread of cGMP and Ca2+. There are a number of factors, however, that could broaden the apparent relationship between functional changes in the light response and the concentration of a diffusible messenger. For these reasons the measured decay length is an upper limit estimate of the spread of adaptation and does not rule out the possibility that Ca2+ and/or cGMP carry the adaptation signal. PMID- 10457082 TI - Kinetic differences at the single molecule level account for the functional diversity of rabbit cardiac myosin isoforms. AB - 1. Cardiac V3 myosin generates slower actin filament velocities and higher average isometric forces (in an in vitro motility assay) when compared with the V1 isoform. 2. To account for differences in V1 and V3 force and motion generation at the molecular level, we characterized the mechanics and kinetics of single V1 and V3 myosin molecules using a dual laser trap setup. 3. No differences in either unitary displacement (approximately 7 nm) or force (approximately 0.8 pN) were observed between isoforms; however, the duration of unitary displacement events was significantly longer for the V3 isoform at MgATP concentrations > 10 microM. 4. Our results were interpreted on the basis of a cross-bridge model in which displacement event durations were determined by the rates of MgADP release from, and MgATP binding to, myosin. 5. We propose that the release rate of MgADP from V3 myosin is half that of V1 myosin without any difference in their rates of MgATP binding; thus, kinetic differences between the two cardiac myosin isoforms are sufficient to account for their functional diversity. PMID- 10457085 TI - Diltiazem causes open channel block of recombinant 5-HT3 receptors. AB - 1. To extend our knowledge of the site and mechanism of action of L-type Ca2+ channel antagonists on 5-HT3 receptors, whole-cell voltage clamp electrophysiology was used to investigate the action of one of these compounds, diltiazem, on the recombinant receptor expressed in human embryonic kidney (HEK) 293 cells. 2. Application of diltiazem with 5-HT (30 microM) caused an increase in the rate of receptor current decay, but did not significantly affect peak current (Ip), the EC50 or the Hill coefficient, indicating a non-competitive mechanism of action. Pre-application of the antagonist had no effect indicating that diltiazem mediates its effects by binding preferentially to the open state of the 5-HT3 receptor. 3. To examine the effects of diltiazem on the open state of the receptor in more detail we used 10 mM 5-hydroxyindole (5-OHi) to reduce receptor desensitisation. These experiments showed that diltiazem causes a rapid, reversible, block in the presence of agonist but can become trapped in the unliganded state of the receptor by prior washout of agonist. Dose-inhibition data yielded an IC50 of 5.5 microM and a Hill coefficient of 0.96; inhibition was slightly voltage dependent as the degree of blockade at +60 mV was reduced. 4. The Hill coefficient of near unity suggests a single molecule of diltiazem mediates inhibition and, indeed, kinetic analysis verified that the interaction of diltiazem with the 5-HT3 receptor was well described by a bimolecular reaction scheme. The results suggest that diltiazem acts by causing open-channel block of the 5-HT3 receptor. PMID- 10457084 TI - Modulation of mammalian dendritic GABA(A) receptor function by the kinetics of Cl and HCO3- transport. AB - 1. During prolonged activation of dendritic GABAA receptors, the postsynaptic membrane response changes from hyperpolarization to depolarization. One explanation for the change in direction of the response is that opposing HCO3- and Cl- fluxes through the GABAA ionophore diminish the electrochemical gradient driving the hyperpolarizing Cl- flux, so that the depolarizing HCO3- flux dominates. Here we demonstrate that the necessary conditions for this mechanism are present in rat hippocampal CA1 pyramidal cell dendrites. 2. Prolonged GABAA receptor activation in low-HCO3- media decreased the driving force for dendritic but not somatic Cl- currents. Prolonged GABAA receptor activation in low-Cl- media containing physiological HCO3- concentrations did not degrade the driving force for dendritic or somatic HCO3- gradients. 3. Dendritic Cl- transport was measured in three ways: from the rate of recovery of GABAA receptor-mediated currents between paired dendritic GABA applications, from the rate of recovery between paired synaptic GABAA receptor-mediated currents, and from the predicted vs. actual increase in synaptic GABAA receptor-mediated currents at progressively more positive test potentials. These experiments yielded estimates of the maximum transport rate (vmax) for Cl- transport of 5 to 7 mmol l-1 s-1, and indicated that vmax could be exceeded by GABAA receptor-mediated Cl- influx. 4. The affinity of the Cl- transporter was calculated in experiments in which the reversal potential for Cl- (ECl) was measured from the GABAA reversal potential in low-HCO3- media during Cl- loading from the recording electrode solution. The calculated KD was 15 mM. 5. Using a standard model of membrane potential, these conditions are demonstrated to be sufficient to produce the experimentally observed, activity-dependent GABA(A) depolarizing response in pyramidal cell dendrites. PMID- 10457086 TI - Two different ionotropic receptors are activated by ATP in rat microglia. AB - 1. Our aim was to assess whether ATP-induced inward currents in microglia are due to a single or more than one purinergic receptor. The ATP dose-response curve showed two components, whose presence might be due to the activation of high and low affinity receptors. 2. The P2Z/P2X7 specific receptor agonist benzoylbenzoyl ATP (Bz-ATP) and some P2 receptor agonists were tested. The rank order of potency was Bz-ATP >> ATP = 2-methylthio-ATP (2-MeSATP) > alpha, beta-methylene ATP (alpha,beta-meATP) >= ADP. beta, gamma-MethyleneATP (beta,gamma-meATP), UTP and adenosine were ineffective. 3. The non-specific P2 receptor antagonist suramin antagonized by 92 +/- 2 % the inward current induced by 100 microM ATP, and by 51 +/- 8 and 68 +/- 6 % those induced by 3 mM ATP and 100 microM Bz-ATP, respectively. The P2Z/P2X7 antagonist oxidized ATP (oATP) almost abolished the inward current induced by 3 mM ATP or Bz-ATP, but was ineffective against 100 microM ATP. 4. Inward currents induced by low ATP concentrations (<= 100 microM) were generally followed by an almost complete and irreversible desensitization, while those elicited by ATP >= 1 mM showed only a partial decline. Interestingly, the inward current induced by 100 microM 2-MeSATP showed a large desensitization, while that induced by Bz-ATP did not. 5. In voltage-ramp experiments, the 100 microM ATP-induced current exhibited a slight inward rectification more visible at negative potentials, while the 3 mM ATP-induced current did not. 6. ATP induced a fast and large increase in [Ca2+] that promptly recovered in the continuous presence of low ATP doses, but did not recover in high ATP doses. As with desensitization, the response to Bz-ATP mimicked that of high doses of ATP. 7. When Ca2+ mobilization due to P2Y receptors was blocked by thapsigargin induced Ca2+ depletion or by pertussis toxin treatment, 10 microM ATP was still able to induce a Ca2+ transient, which represented the contribution of the Ca2+ influx induced by P2X receptors 8. In conclusion, the inward currents and a fraction of the Ca2+ transients induced by ATP in microglia are due to at least two ATP-sensitive receptor channel types, whose different properties and sensitivity to ATP may be associated with different functional roles. PMID- 10457088 TI - Electrophysiological properties of electrical synapses between rat sympathetic preganglionic neurones in vitro. AB - 1. The electrophysiological properties of electrical synaptic transmission between sympathetic preganglionic neurones (SPNs) in slices of rat spinal cord were investigated using simultaneous dual-electrode patch-clamp recordings. Electrotonic coupling was directly demonstrated between 21 pairs of SPNs. 2. Coupling coefficients determined from the steady-state response of both neurones to current steps injected into either neurone ranged from 0. 02 to 0.48 (0.18 +/- 0.02, mean +/- s.e.m.). Synapses were bidirectional and symmetrical for the majority of connections with coupling coefficients similar in either direction. Asymmetrical coupling between a minority of cell pairs was due to differences in passive neuronal properties rather than rectification of the synaptic conductances. 3. Action potentials were manifest in adjoining cells as biphasic electrical postsynaptic potentials (ePSPs), composed of a rapid depolarising component followed by a more prolonged hyperpolarisation with amplitudes of 1.2 +/- 0.2 and 2.1 +/- 0.6 mV, respectively. 4. Postsynaptic potentials resembled low-pass filtered presynaptic spikes with frequency dependence determined by the junctional conductance and postsynaptic membrane properties. Increases in presynaptic action potential frequency caused attenuation of the hyperpolarising component of the ePSP that was attributed to shorter duration presynaptic spikes being more markedly filtered. 5. Synchronisation of spontaneous action potentials between electrotonically coupled neurones was driven by subthreshold membrane potential activity resembling repetitive ePSPs. Synchronous spike firing in previously silent neurones could be driven by suprathreshold ePSPs induced by suprathreshold depolarisation of a single adjoining neurone. 6. These data characterise reliable communication of sub- and suprathreshold activity by electrical synapses enabling synchronised SPN firing which may contribute to generation of coherent sympathetic rhythms and promote summation of inputs to postganglionic neurones. PMID- 10457087 TI - Control of apical membrane chloride permeability in the renal A6 cell line by nucleotides. AB - 1. The effect of extracellular nucleotides applied on the apical side of polarised A6 cells grown on permeant filters was investigated by measuring the changes in (i) the 36Cl efflux through the apical membranes, (ii) the intracellular chloride concentrations (aCli, measured with N-(6-methoxyquinolyl) acetoethyl ester, MQAE), (iii) ICl, the short-circuit current in the absence of Na+ transport and (iv) the characteristics of the apical chloride channels using a patch-clamp approach. 2. ATP or UTP (0.1-500 microM) transiently stimulated ICl. The sequence of purinergic agonist potencies was UTP = ATP > ADP >> the P2X selective agonist beta,gamma-methylene ATP = the P2Y-selective agonist 2 methylthioATP. Suramin (100 microM) as the P2Y antagonist Reactive Blue 2 (10 microM) had no effect on the UTP (or ATP)-stimulated current. These findings are consistent with the presence of P2Y2-like receptors located on the apical membranes of A6 cells. Apical application of adenosine also transiently increased ICl. This effect was blocked by theophylline while the UTP-stimulated ICl was not. The existence of a second receptor, of the P1 type is proposed. 3. ATP (or UTP)-stimulated ICl was blocked by apical application of 200 microM N phenylanthranilic acid (DPC) or 100 microM niflumic acid while 100 microM glibenclamide was ineffective. 4. Ionomycin and thapsigargin both transiently stimulated ICl; the nucleotide stimulation of ICl was not suppressed by pre treatment with these agents. Chlorpromazin (50 microM), a Ca2+-calmodulin inhibitor strongly inhibited the stimulation of ICl induced either by apical UTP or by ionomycin application. BAPTA-AM pre-treatment of A6 cells blocked the UTP stimulated ICl. Niflumic acid also blocked the ionomycin stimulated ICl. 5. A fourfold increase in 36Cl effluxes through the apical membranes was observed after ATP or UTP application. These increases of the apical chloride permeability could also be observed when following aCli changes. Apical application of DPC (1 mM) or 5-nitro-2(3-phenylpropylamino)benzoic acid (NPPB; 500 microM) produced an incomplete inhibition of 36Cl effluxes through the apical membranes in ATP stimulated and in untreated monolayers. 6. In single channel patch-clamp experiments, an apical chloride channel with a unitary single channel conductance of 7.3 +/- 0.6 pS (n = 12) was usually observed. ATP application induced the activation of one or more of these channels within a few minutes. 7. These results indicate that multiple purinergic receptor subtypes are present in the apical membranes of A6 cells and that nucleotides can act as modulators of Cl- secretion in renal cells. PMID- 10457089 TI - Acid-evoked quantal catecholamine secretion from rat phaeochromocytoma cells and its interaction with hypoxia-evoked secretion. AB - 1. Amperometric recordings using polarized carbon fibre microelectrodes were used to detect exocytosis of catecholamines from rat phaeochromocytoma (PC12) cells in response to a reduction in pHo. 2. Exocytosis was detected at pHo levels of between 7.2 and 6.8. This was probably due to intracellular acidification, since acid-evoked secretion was enhanced by the Na+-H+ exchange blocker ethylisopropylamiloride (30 microM), and was mimicked by sodium propionate (10 mM), which causes selective intracellular acidosis. 3. Acid-evoked exocytosis was abolished by removal of Ca2+o or application of 200 microM Cd2+. It was unaffected by nifedipine, but significantly reduced by either omega-conotoxin GVIA (1 microM) or omega-agatoxin GIVA (200 nM). The two toxins applied together almost completely abolished (> 97 %) acid-evoked secretion. 4. Hypoxia-evoked catecholamine release was potentiated under acidic conditions and suppressed under alkaline conditions in a manner which indicated a greater than additive interaction of these two stimuli. 5. Our results indicate that, like carotid body arterial chemoreceptors, PC12 cells represent model chemoreceptor cells for both hypoxia and acidity and that the release of catecholamines in response to these physiological stimuli is dependent on Ca2+ influx through voltage-gated N- and P/Q-type Ca2+ channels. PMID- 10457090 TI - Dynamics of primate P retinal ganglion cells: responses to chromatic and achromatic stimuli. AB - 1. The majority of primate retinal ganglion cells (RGCs) project to the parvocellular layers of the lateral geniculate nucleus (LGN). These P cells play a central role in early visual processing. 2. An improved method of systems analysis has allowed us to explore the dynamics of the colour-opponent subregions of P-cell receptive fields with a single chromatic stimulus. The data show that the centre and surround subregions of the P-cell receptive field have similar temporal responses, but the surround is slightly delayed. The centre and surround demonstrate a large degree of chromatic selectivity. 3. The responses of the centre and surround subregions were fitted with a linear model and the model was used to predict the responses of P cells to new chromatic and achromatic stimuli. Although linear models predict the chromatic responses well, simple linear combinations of centre and surround responses fail to predict P-cell responses to achromatic stimuli. 4. The temporal responses of the different subpopulations of P cells, such as ON/OFF or L-centre/M-centre were not significantly different. PMID- 10457091 TI - Expression density and functional characteristics of the outer hair cell motor protein are regulated during postnatal development in rat. AB - 1. The non-linear capacitance (Cnon-lin) of postnatal outer hair cells (OHCs) of the rat was measured by a patch-clamp lock-in technique. Cnon-lin is thought to result from a membrane protein that provides the molecular basis for the unique electromotility of OHCs by undergoing conformational changes in response to changes in membrane potential (Vm). Protein conformation is coupled to Vm by a charged voltage sensor, which imposes Cnon-lin on the OHC. Cnon-lin was investigated in order to characterize the surface expression and voltage dependence of this motor protein during postnatal development. 2. On the day of birth (P0), Cnon-lin was not detected in OHCs of the basal turn of the cochlea, whilst it was 89 fF in apical OHCs. Cnon-lin increased gradually during postnatal development and reached 2.3 pF (basal turn, P9) and 7.5 pF (apical turn, P14) at the oldest developmental stages covered by our measurements. The density of the protein in the plasma membrane, deduced from non-linear charge movement per membrane area, increased steeply between P6 and P11 and reached steady state (4200 e- microm-2) at about P12. 3. Voltage at peak capacitance (V) shifted with development from hyperpolarized potentials shortly after birth (-88.3 mV, P2) to the depolarized potential characteristic of mature OHCs (-40.8 mV, P14). This developmental difference in V was also observed in outside-out patches immediately after patch excision. During subsequent wash-out V shifted towards the depolarized value found in the adult state, suggesting a direct modulation of the molecular motor. 4. Thus, the density of the motor protein in the plasma membrane and also its voltage dependence change concomitantly in the postnatal period and reach adult characteristics right at the onset of hearing. PMID- 10457092 TI - Adaptation of the odour-induced response in frog olfactory receptor cells. AB - 1. Receptor current and spiking responses were recorded simultaneously from isolated frog olfactory receptor cells using the suction pipette technique. Cells were stimulated with the odour cineole by rapid exchange of the solution bathing the olfactory cilia. 2. The receptor current response to a 1 s odour stimulus increased in a graded manner over a 300-fold range of odour concentration without clear saturation, and was accompanied by a train of action potentials. As the concentration of the odour stimulus increased, the frequency of firing increased also, until it saturated at the highest concentrations. The number of spikes evoked by the stimulus first increased and then decreased with increasing concentration, reaching a maximum at intermediate odour concentrations. The dose response relation for spike firing rose at lower odour concentrations than the dose-response relation for the receptor current response. 3. Adaptation to steady odour stimuli was investigated by exposing the cilia to a 4 s odour pre-pulse and then to a 1 s odour test pulse. As the pre-pulse concentration was increased the dose-response relations derived from the receptor current and spiking responses shifted to higher absolute test pulse concentrations. However the number of spikes fired in response to a given test pulse was little affected by the pre pulse until, at the highest pre-pulse concentrations spike firing was abolished despite the continued presence of a receptor current response. 4. The sensitivity of the receptor-current response to incremental stimuli fell with increasing pre pulse concentration, declining with a limiting slope of 2.4 in double logarithmic co-ordinates. The sensitivity determined from the spiking responses declined to zero at a lower pre-pulse concentration, reflecting the abolition of spike firing at pre-pulse concentrations which still evoked a graded receptor-current response. PMID- 10457093 TI - High intracellular [Ca2+] alters sarcoplasmic reticulum function in skinned skeletal muscle fibres of the rat. AB - 1. The effect on sarcoplasmic reticulum (SR) function of exposure to high intracellular [Ca2+] was studied in mechanically skinned fibres from the extensor digitorum longus muscle of the rat, using caffeine to assay the SR Ca2+ content. 2. A 15 s exposure to 50 microM Ca2+ irreversibly reduced the ability of the SR to load/retain Ca2+ and completely abolished depolarization-induced Ca2+ release, whereas a 90 s exposure to 10 microM Ca2+ had no detectable effect on either function. The reduction in net SR Ca2+ uptake: (a) was near-maximal with treatment at 50 microM Ca2+, (b) was unrelated to voltage-sensor function, and (c) persisted unchanged for > 20 min. The reduction was primarily due to a threefold increase in leakage of Ca2+ out of the SR. This increased leakage was not substantially blocked by the presence of 10 mM Mg2+ or 2 microM Ruthenium Red. 3. The adverse effect on SR function of exposure to high [Ca2+] could also be observed by the reduction in the ability of the SR to maintain a low [Ca2+] within the skinned fibre in the face of elevated [Ca2+] in the bathing solution. When bathed in a solution with approximately 1.5 microM Ca2+ (0.75 mM CaEGTA EGTA), skinned fibres produced only low force responses for many minutes, but after high [Ca2+] treatment (15 s exposure to 50 microM Ca2+) they showed large, steady or oscillatory force responses. 4. These findings indicate that, in addition to uncoupling the Ca2+ release channels from the voltage sensors, exposure of skinned fibres to high [Ca2+] causes a persistent increase in resting Ca2+ efflux from the SR. Such efflux in an intact fibre would alter the distribution of Ca2+ between the SR, the cytoplasm and the extracellular solution. These results may be relevant to the basis of low-frequency fatigue and possibly other conditions in muscle. PMID- 10457094 TI - Actin polymerization stimulated by contractile activation regulates force development in canine tracheal smooth muscle. AB - 1. The role of actin polymerization in the regulation of smooth muscle contractility was investigated in canine trachealis muscle strips. The effect of contractile activation on the content of monomeric globular (G)-actin was estimated by the method of DNase I inhibition. The G-actin content was 30 % lower in extracts of muscle strips activated with 10-4 M acetylcholine (ACh) than in extracts from unstimulated muscle strips. The decrease in G-actin in response to contractile stimulation was prevented by latrunculin-A, an agent that prevents actin polymerization by binding to G-actin monomers. 2. The inhibition of actin polymerization by latrunculin-A markedly depressed force development in response to ACh but had no effect on ACh-induced myosin light chain (MLC) phosphorylation. Latrunculin also suppressed the length sensitivity of force during ACh-induced isometric contractions. The actin-capping agent cytochalasin-D also markedly inhibited force and caused only a slight decrease in MLC phosphorylation. Cytochalasin-D also inhibited force in alpha-toxin-permeabilized muscle strips that were activated either by Ca2+ or by ACh at constant pCa. No disorganization of smooth muscle cell ultrastructure was detected by electron microscopy or by immunofluorescence microscopy of muscles treated with either agent. 3. The results suggest that the polymerization of actin is stimulated by the contractile activation of tracheal smooth muscle and that this actin polymerization contributes directly to force development. In addition, actin filament remodelling contributes to the length sensitivity of tracheal smooth muscle contractility. PMID- 10457095 TI - Relationship between transient outward K+ current and Ca2+ influx in rat cardiac myocytes of endo- and epicardial origin. AB - 1. The transient outward K+ current (Ito) is a major repolarizing ionic current in ventricular myocytes of several mammals. Recently it has been found that its magnitude depends on the origin of the myocyte and is regulated by a number of physiological and pathophysiological signals. 2. The relationship between the magnitude of Ito, action potential duration (APD) and Ca2+ influx (QCa) was studied in rat left ventricular myocytes of endo- and epicardial origin using whole-cell recordings and the action potential voltage-clamp method. 3. Under control conditions, in response to a depolarizing voltage step to +40 mV, Ito averaged 12.1 +/- 2.6 pA pF-1 in endocardial (n = 11) and 24.0 +/- 2.6 pA pF-1 in epicardial myocytes (n = 12; P < 0.01). APD90 (90 % repolarization) was twice as long in endocardial myocytes, whereas QCa inversely depended on the magnitude of Ito. L-type Ca2+ current density was similar in myocytes from both regions. 4. To determine the effects of controlled reductions of Ito on QCa, recordings were repeated in the presence of increasing concentrations of the Ito inhibitor 4 aminopyridine. 5. Inhibition of Ito by as little as 20 % more than doubled QCa in epicardial myocytes, whereas it had only a minor effect on QCa in myocytes of endocardial origin. Further inhibition of Ito led to a progressive increase in QCa in epicardial myocytes; at 90 % inhibition of Ito, QCa was four times larger than the control value. 6. We conclude that moderate changes in the magnitude of Ito strongly affect QCa primarily in epicardial regions. An alteration of Ito might therefore allow for a regional regulation of contractility during physiological and pathophysiological adaptations. PMID- 10457097 TI - Identification of the cells underlying pacemaker activity in the guinea-pig upper urinary tract. AB - 1. The varying profile of cell types along the muscle wall of the guinea-pig upper urinary tract was examined electrophysiologically, using intracellular microelectrodes, and morphologically, using both electron and confocal microscopy. 2. Simple 'pacemaker' oscillations (frequency of 8 min-1) of the membrane potential were recorded in both the pelvi-calyceal junction (83 % of cells) and the proximal renal pelvis (15 % of cells), but never in the distal renal pelvis or ureter. When filled with the cell marker, neurobiotin, 'pacemaker' cells were spindle shaped and approximately 160 microm in length. 3. In most cells of the ureter (100 %) and in both the proximal (75 %) and distal (89 %) renal pelvis, spontaneous action potentials (frequency of 3-5 min-1) consisted of an initial spike, followed by a number of potential oscillations superimposed on a plateau phase. When filled with neurobiotin, cells firing these 'driven' action potentials, were spindle shaped and > 250 microm in length. 4. Greater than 80 % of smooth muscle cells in the pelvi-calyceal junction were 'atypical', having < 40 % of their sectional areas occupied by loosely packed contractile filaments. Most of the smooth muscle cells in the ureter (99.7 %) and both the proximal (83 %) and distal (97.5 %) renal pelvis were of 'typical' appearance in that they contained cytoskeletal and contractile elements occupying > 60 % of cross-sectional area. 5. A third type of spontaneously discharging cell fired 'intermediate' action potentials (3-4 min-1), consisting of a single spike followed by a quiescent plateau and an abrupt repolarization. These cells were morphologically similar to interstitial cells of Cajal (ICC). However, these 'ICC like' cells were not immuno-reactive for c-Kit, the proto-oncogene for tyrosine kinase. 6. In summary, 'atypical' smooth muscle cells were predominant in the pelvi-calyceal junction and fired 'pacemaker' potentials at a frequency significantly higher than 'driven' action potentials recorded in 'typical' smooth muscle cells throughout the renal pelvis and ureter. 'Intermediate' action potentials were recorded in 'ICC-like' cells in both the pelvi-calyceal junction and renal pelvis. We suggest that these 'ICC-like' cells act as a preferential pathway, conducting and amplifying pacemaker signals to initiate action potential discharge in the driven areas of the upper urinary tract. PMID- 10457096 TI - Tetracaine can inhibit contractions initiated by a voltage-sensitive release mechanism in guinea-pig ventricular myocytes. AB - 1. Effects of tetracaine on membrane currents and cell shortening were measured with high resistance electrodes, single-electrode voltage clamp (switch clamp) and a video edge detector at 37 C in cardiac ventricular myocytes. 2. Sequential voltage steps from -65 mV to -40 and 0 mV were used to activate two mechanisms of excitation-contraction (EC) coupling separately. The step to -40 mV activated the voltage-sensitive release mechanism (VSRM); the step to 0 mV1 activated Ca2+ induced Ca2+ release (CICR) coupled to inward Ca2+ current (IL). 3. Exposure to 100-300 microM tetracaine inhibited VSRM contractions but not CICR contractions. Inhibition of VSRM contractions was independent of INa blockade. In contrast, 100 microM Cd2+ blocked IL and CICR contractions, but not VSRM contractions. Simultaneous application of both agents blocked both mechanisms of EC coupling. 4. Contraction-voltage relationships were sigmoidal when the VSRM was available. However, when the VSRM was inhibited with 100-300 microM tetracaine, contraction voltage relationships became bell-shaped. The tetracaine-insensitive contractions were abolished by 0.1 microM ryanodine, indicating that they were dependent on release of SR Ca2+. 5. At a higher concentration (1 mM) tetracaine also inhibited IL and contractions triggered by IL; however, the time course of effects on IL and associated contractions were different than for VSRM contractions. 6. With continuous application of tetracaine, the VSRM remained inhibited although SR Ca2+ stores increased 4-fold as assessed with caffeine. CICR contractions were not inhibited and maximum amplitude of contraction was not reduced. 7. Rapid application of tetracaine just before and during test steps also inhibited VSRM contractions, but without significantly affecting sarcoplasmic reticulum (SR) Ca2+ stores or CICR contractions. Maximum amplitude of contraction was reduced. 8. Rapid application of tetracaine (100-300 microM) allows preferential inhibition of the VSRM and provides a pharmacological method to assess the contribution of the VSRM to EC coupling. PMID- 10457098 TI - Modulation of mammary development and programmed cell death by the frequency of milk removal in lactating goats. AB - 1. Unilateral changes in mammary cell number are elicited when one gland is milked more or less frequently than the contralateral gland in lactating goats. These changes were investigated using histochemical and immunocytochemical markers of mammary cell types, and the degree of mammary apoptosis was determined by end-labelling of fragmented DNA. 2. Histological analysis confirmed that unilateral cessation of milking initiated involution and cell loss preferentially in the unmilked gland. The presence of fragmented DNA and morphological characteristics consistent with apoptosis demonstrated that these changes in mammary cell number in unmilked glands were, in part, the result of programmed alveolar cell death. 3. De-differentiation of the remaining secretory cells to ductal epithelial cells occurred with an increase in staining of cytokeratin markers and decreased staining by peanut lectin and casein antisera. 4. Differential once- and thrice-daily milking of lactating goats was also associated with unilateral changes in mammary cell number and milk yield. Milk yield and alveolar size were reduced after 4 weeks of infrequent milking. The latter was due to the increased loss of secretory cells by apoptosis, as indicated by a higher degree of fragmented DNA laddering. 5. After 10 weeks of differential milking, a homogeneous secretory morphology, albeit with smaller alveoli, was maintained in thrice-daily milked glands. Once-daily milked glands possessed a heterogeneous composition of terminal structures, resulting in the simultaneous presence of secretory and involuting alveoli as well as resting ductules. 6. The differences in programmed cell death and mammary morphology between unmilked and twice-daily milked glands, and between once- and thrice daily milked glands, suggests that mammary apoptosis is subject to modulation by intra-mammary mechanisms sensitive to the frequency of milk removal. PMID- 10457099 TI - In vivo ATP synthesis rates in single human muscles during high intensity exercise. AB - 1. In vivo ATP synthesis rates were measured in the human medial gastrocnemius muscle during high intensity exercise using localized 31P-magnetic resonance spectroscopy (31P-MRS). Six-second localized spectra were acquired during and following a 30 s maximal voluntary rate exercise using a magnetic resonance image guided spectral localization technique. 2. During 30 s maximal voluntary rate exercise, ATPase fluxes were predominantly met by anaerobic ATP sources. Maximal in vivo glycogenolytic rates of 207 +/- 48 mM ATP min-1 were obtained within 15 s, decreasing to 72 +/- 34 mM ATP min-1 by the end of 30 s. In contrast, aerobic ATP synthesis rates achieved 85 +/- 2 % of their maximal capacity within 9 s and did not change throughout the exercise. The ratio of peak glycolytic ATP synthesis rate to maximal oxidative ATP synthesis was 2.9 +/- 0.9. 3. The non-Pi, non-CO2 buffer capacity was calculated to be 27.0 +/- 6. 2 slykes (millimoles acid added per unit change in pH). At the cessation of exercise, Pi, phosphomonoesters and CO2 were predicted to account for 17.2 +/- 1.5, 5.57 +/- 0.97 and 2.24 +/- 0.34 slykes of the total buffer capacity. 4. Over the approximately linear range of intracellular pH recovery following the post exercise acidification, pHi recovered at a rate of 0.19 +/- 0.03 pH units min-1. Proton transport capacity was determined to be 16.4 +/- 4.1 mM (pH unit)-1 min-1 and corresponded to a maximal proton efflux rate of 15.3 +/- 2.7 mM min-1. 5. These data support the observation that glycogenolytic and glycolytic rates are elevated in vivo in the presence of elevated Pi levels. The data do not support the hypothesis that glycogenolysis follows Michealis-Menten kinetics with an apparent Km for [Pi] in vivo. 6. In vivo -measured ATP utilization rates and the initial dependence on PCr and glycolysis were similar to those previously reported in in situ studies involving short duration, high intensity exercise. This experimental approach presents a non-invasive, quantitative measure of peak glycolytic rates in human skeletal muscle. PMID- 10457100 TI - Effect of adrenaline on glucose kinetics during exercise in adrenalectomised humans. AB - 1. The role of adrenaline in regulating hepatic glucose production and muscle glucose uptake during exercise was examined in six adrenaline-deficient, bilaterally adrenalectomised humans. Six sex- and age-matched healthy individuals served as controls (CON). 2. Adrenalectomised subjects cycled for 45 min at 68 +/ 1 % maximum pulmonary O2 uptake (VO2,max), followed by 15 min at 84 +/- 2 % VO2, max without (-ADR) or with (+ADR) adrenaline infusion, which elevated plasma adrenaline levels (45 min, 4.49 +/- 0.69 nmol l-1; 60 min, 12.41 +/- 1.80 nmol l 1; means +/- s.e.m.). Glucose kinetics were measured using [3-3H]glucose. 3. Euglycaemia was maintained during exercise in CON and -ADR, whilst in +ADR plasma glucose was elevated. The exercise-induced increase in hepatic glucose production was similar in +ADR and -ADR; however, adrenaline infusion augmented the rise in hepatic glucose production early in exercise. Glucose uptake increased during exercise in +ADR and -ADR, but was lower and metabolic clearance rate was reduced in +ADR. 4. During exercise noradrenaline and glucagon concentrations increased, and insulin and cortisol concentrations decreased, but plasma levels were similar between trials. Adrenaline infusion suppressed growth hormone and elevated plasma free fatty acids, glycerol and lactate. Alanine and beta-hydroxybutyrate levels were similar between trials. 5. The results demonstrate that glucose homeostasis was maintained during exercise in adrenalectomised subjects. Adrenaline does not appear to play a major role in matching hepatic glucose production to the increase in glucose clearance. In contrast, adrenaline infusion results in a mismatch by simultaneously enhancing hepatic glucose production and inhibiting glucose clearance. PMID- 10457102 TI - Cross-reactivity between milk proteins of different animals. PMID- 10457101 TI - Gas cooking appliances and indoor pollution. PMID- 10457103 TI - Exocytotic events in eosinophils and mast cells. PMID- 10457104 TI - The genetic basis of atopic asthma. PMID- 10457105 TI - Effects of exposure to gas cooking in childhood and adulthood on respiratory symptoms, allergic sensitization and lung function in young British adults. AB - BACKGROUND: There is evidence that people who use gas for cooking have reduced lung function and experience more respiratory symptoms than those who use other fuels for cooking. OBJECTIVES: To study the effect of the presence of a gas cooker in the home, during both childhood and adulthood, on respiratory symptoms, allergic sensitization and ventilatory function among young adults. METHODS: A sample of 1449 young adults born in Britain 3-9 March 1958, who have been followed from birth to ages 7, 11, 16, 23 and 33 years, were examined at home at age 34-35 years. FEV1 and FVC were measured before and 20 min after inhalation of 400 microg salbutamol, and skin prick tests performed with three allergen extracts (grass, Der p 1 and cat). An interview on respiratory symptoms and indoor environmental exposures was included. RESULTS: No association was found between gas cooking in childhood or adulthood and incidence or prognosis of asthma/wheeze, allergic sensitization or current severity of respiratory symptoms. Subjects who currently used gas for cooking had a significantly reduced FEV1 (- 70 mL, 95% CI +/- 56) but not FVC (- 35 mL, 95% CI +/- 61) compared with those who used electricity for cooking. This reduction in FEV1 was concentrated among men and current asthmatics. CONCLUSION: The use of gas for cooking is unlikely to be a major influence on respiratory morbidity in young adults. PMID- 10457106 TI - Elevated cord serum IgE increases the risk of aeroallergen sensitization without increasing respiratory allergic symptoms in early childhood. AB - BACKGROUND: Increasing prevalence of allergic disorders has focused attention on primary prevention. There is a need to improve the accuracy of early-life predictors of atopy so that the at-risk population can be accurately defined and preventive measures instituted. OBJECTIVE: The predictive capacity of elevated cord IgE, with or without family history of atopy, to allergic symptoms and skin prick test (SPT) sensitization is evaluated in a birth cohort followed up prospectively for 4 years. METHODS: A birth cohort of 1456 consecutively born children was recruited in 1989. Data were collected on family history of atopy and cord serum total IgE (cord IgE) was measured. Of these, 1218 children were seen in the clinic at 4 years to determine the development of symptoms and signs of allergic disease and 981 were skin tested to a range of common food and aeroallergens. RESULTS: Of 1218 children reviewed at age 4 years, 218 (17.8%) had symptoms of respiratory allergy and, of those skin tested (n = 981), 192 (19.6%) reacted positively. Twice as many children with elevated cord IgE (>/= 0.5 kU/L) at birth became sensitized to aeroallergens by age 4 years (34.8% vs 17.3%, P < 0. 001). Positive predictive value (PPV) of elevated cord IgE for the development of aeroallergen sensitization was better than that of family history of atopy (34.8 vs 22.6%). Combining paternal atopy with elevated cord IgE substantially increased the predictive capacity (PPV 77.8%). Cord IgE levels did not correlate with clinical asthma or rhinitis at age 4 years and PPV for allergic respiratory symptoms remained poor at all cutoffs. CONCLUSION: Cord IgE is better than family history for predicting atopy as defined by allergen sensitization and this predictive value can be further increased by combining cord IgE with paternal atopy. PMID- 10457107 TI - Genetic linkage of HLA-class II locus to mite-specific IgE immune responsiveness. AB - BACKGROUND: IgE response to common inhalant allergens seems to be the major determinant of the development of atopic rhinitis and asthma but it has been difficult to demonstrate genetic control of the IgE response. OBJECTIVE: To investigate genetic linkage between specific IgE reactions to purified aero allergens (grass, birch, cat, mite) and the HLA-class II locus. METHODS: DNA based HLA-class II typing was performed for determination of DRB1, DQB1 and DPB1 alleles. Linkage was studied by the affected sibpair method and the extended transmission disequilibrium test in 100 children from 40 nuclear families selected from a homogeneous population in south-western Germany. RESULTS: Significant linkage of mite-specific IgE response to HLA-DPB (P = 0.00001), HLA DRB (0.02) and HLA-DQB (P = 0.001) was revealed by sibpair analysis of MHC class II alleles and confirmed by the extended transmission disequilibrium test for HLA DRB (P = 0.01) and HLA-DPB (P = 0.04). No consistent significant linkage between the HLA-class II locus and IgE response to grass pollen, birch pollen, and cat dander could be demonstrated. CONCLUSION: The findings are consistent with the existence of one or more genes in the HLA-class II region modifying the IgE immune response to common environmental allergens. PMID- 10457108 TI - Allergy to bovine beta-lactoglobulin: specificity of human IgE to tryptic peptides. AB - BACKGROUND: Bovine beta-lactoglobulin (Blg) is a major cow's milk allergen. It is the main whey protein, without any counterpart in human milk. Blg chemical hydrolysates appeared to retain most of the immunoreactivity of the native protein. Allergenicity of Blg has already been shown to be associated with the four peptides derived from cyanogen bromide cleavage of Blg. OBJECTIVES: To map the major allergenic epitopes (e.g. regions of the molecule able to bind IgE) on Blg using specific IgE from sera of 46 milk-allergic patients as a probe. METHODS: Direct and competitive inhibition enzyme immunoassays involving immobilized native protein or purified peptides derived from Blg tryptic cleavage. RESULTS: Several peptides capable of specifically binding human IgEs were identified and were classified according to the intensity and frequency of the responses. The major epitopes appeared to be fragments (41-60), (102-124) and (149-162) recognized by 92, 97 and 89% of sera, respectively, whilst a second group which contained the fragments (1-8) and (25-40) was recognized by 58 and 72% of the population. A third group, comprising peptides (9-14), (84-91) and (92 100), was still detected by more than 40% of sera. CONCLUSION: Three peptides were identified as major epitopes, recognized by a large majority of human IgE antibodies. Numerous other epitopes are scattered all along the Blg sequence. PMID- 10457109 TI - Allergy to goat and sheep cheese with good tolerance to cow cheese. AB - BACKGROUND: We report on a patient who experienced allergic reactions after eating goat cheese and after touching goat and sheep cheese, but not after consuming cow's milk dairy products. OBJECTIVE: To assess the allergenicity and IgE-binding capacity of the caseins from the three different species. METHODS: Skin prick tests were carried out using whole milk and caseins from three different species (goat, sheep and cow), and whey fractions of cow's milk. Total serum IgE and specific IgE to cow's milk proteins were measured by CAP system and specific IgE against caseins and whole milk were determined by ELISA technique. To evaluate allergenic cross-reactivity, inhibition of the IgE ELISA activity to goat's milk and goat casein was tested for the three caseins. SDS-PAGE and immunoblotting was used to determine IgE binding bands in caseins. RESULTS: Skin tests were positive to sheep and goat's milk, sheep and goat casein, as well as to sheep and goat cheese. Total serum IgE was 66 kU/L and IgE determinations by CAP were negative. IgE ELISA against the caseins from goat and sheep was strongly positive, whereas it was negative to cow casein. ELISA inhibition assays revealed a high degree of cross-reactivity between goat casein and sheep casein. Immunoblotting showed three IgE-binding bands in goat casein at 31, 27 and 22 kDa, which may correspond to alpha-, beta- and gamma-caseins. A band at about 31 kDa was observed in sheep casein and another band at 34 kDa was recognized in cow casein. CONCLUSION: This patient developed allergy to goat and sheep cheese with good tolerance to cow's milk. We identified goat casein as the main allergen causing sensitization in this patient as demonstrated by in vivo and in vitro tests. A high degree of cross-reactivity between goat and sheep casein was observed. PMID- 10457110 TI - Epidemiological study of the prevalence of allergic reactions to Hymenoptera in a rural population in the Mediterranean area. AB - BACKGROUND: Systemic allergic reactions to Hymenoptera venom occur in a percentage that varies from 0.4 to 3.3%. Epidemiological studies indicate that from 15 to 25% of the general population can be sensitized to different Hymenoptera venom as well as the fact that the degree of exposure may be related to the prevalence found in those studies. OBJECTIVE: The objective of this study was to evaluate the prevalence of insect sting allergy and the venom sensitization in a rural population to three Hymenoptera previously found in the area: Polistes dominulus (Pd), Vespula germanica (Vg) and honey bee (Hb). METHODS: A rural community located in the south-east of Spain, close to the Mediterranean Sea, was selected since the stinging Hymenoptera having been previously identified. A random sample of 310 subjects from the village census was studied. A questionnaire and a serum sample were obtained from every patient. The evaluation was conducted by a family doctor, who focused on the reactions to Hymenoptera sting, age, sex, occupation, atopia, previous Hymenoptera sting, stinging insect, interval to last sting and average stings per year. RAST to Hymenoptera venoms were also determined. RESULTS: The prevalence of systemic reactions was 2.3% (57.6% of them had a positive RAST). Large local reactions were found in 26.4% (only 28.5% of them had a positive RAST). Asymptomatic sensitization (positive RAST) was observed in 16.4% of subjects without reaction. Only a weak correlation between subjects with less than 3 years' interval to last sting exposure and positive RAST results was noted, whether they presented with a clinical reaction or not (P < 0.05). CONCLUSIONS: The prevalence of systemic sting reactions in our rural community is higher than other general populations in the same Mediterranean area, and similar to other rural populations studied. The degree of exposure influences not only the prevalence found but also the detection of specific serum IgE. PMID- 10457111 TI - Serological characterization of allergens in poppy seeds. AB - BACKGROUND AND OBJECTIVE: Poppy seeds in food can induce immediate-type allergic reactions ranging from mild local symptoms to severe anaphylactic reactions. Previous publications showed that poppy seeds cross-react with other plant derived allergens. The IgE-binding components have not been defined so far. METHODS: We analysed sera from 11 patients with adverse reactions after ingestion of poppy seed-containing food by IgE-immunoblotting. Nine of 11 patients showed concomitant IgE binding to allergens of birch, mugwort or grass pollen in RAST CAP, and suffered from characteristic seasonal symptoms. RESULTS: Ten of 11 patients showed IgE binding to a 45-kDa protein, 4/11 to a 34-kDa, 5/11 to a 17 kDa, 5/11 to a 14-kDa, and 3/11 to a 5-kDa component. Furthermore, individual IgE binding to proteins of 20, 25, 30 and 40 kDa proteins could be observed. Periodate treatment of blots markedly reduced the IgE binding capacity of the 40- and 45-kDa compounds, indicating the existence of IgE epitopes of the carbohydrate type. Inhibition studies indicated the presence of homologues of pollen allergens in extracts from poppy seeds, i.e. Bet v 1 and Bet v 2. CONCLUSION: The serological analysis showed IgE binding to protein and sugar components of poppy seeds. The 40- and 45-kDa allergens are glycoproteins and contain IgE binding carbohydrate moieties. Moreover, cross-reacting homologues of pollen allergens including Bet v 1 and profilin were detected in poppy seed extract. PMID- 10457112 TI - Identification and characterization of the major allergen of the Humulus japonicus pollen. AB - BACKGROUND: Pollen of Humulus japonicus has been known as one of the important causes of pollinosis in Korea and China. To date, the major allergen of H. japonicus has not been determined. OBJECTIVE: To identify the major allergen of H. japonicus pollen and characterize its biochemical properties. METHODS: With the sera of 29 patients reactive to H. japonicus, the major allergen of H. japonicus was determined from the results of IgE immunoblotting and ELISA inhibition. The biochemical properties of the major allergen of H. japonicus were evaluated by lectin blotting assay and 2-dimensional PAGE blot. N-terminal amino acid sequences were determined by the Edman degradation method. The suggested major allergen was purified by DEAE anion exchange and gel filtration chromatography. RESULTS: Twenty-nine sera contained IgE bound to the 10, 16, 20, 29 and 42 kDa proteins of H. japonicus in immunoblot analysis. A protein of 10 kDa was the most prevalent allergen in the sera of H. japonicus-reactive patients (72%). The ELISA optical density of H. japonicus-specific IgE was not inhibited by pollen extracts of birch, oak, rye grass and mugwort. The 10-kDa allergen was neither stained with PAS nor bound with ConA and five other lectins. The isoelectric point of the 10-kDa allergen was approximately pH 5.1. We sequenced the N-terminal amino acids of the 10-kDa allergen, which was not homologous with any previously characterized allergen. The 10-kDa allergen could be purified with DEAE anion exchange and gel filtration chromatography. Maximum inhibitions of H. japonicus-specific IgE ELISA by whole extract of H. japonicus and purified 10-kDa allergen were more than 97 and 88%, respectively, while the 50% inhibitory concentration of the whole extract of H. japonicus and purified 10 kDa were 38 and 20 ng/mL, respectively. CONCLUSION: The 10-kDa peptide could be a major allergen of H. japonicus. Its isoelectric point was 5.1 and it did not bind with lectins. The N-terminal amino acid sequence of the 10-kDa major allergen was also determined. PMID- 10457113 TI - Role of carbohydrate moieties in IgE binding to allergenic components of Cupressus arizonica pollen extract. AB - BACKGROUND: A reduction of IgE immunoreactivity after periodate-treatment has been previously reported for various glycoprotein allergens. OBJECTIVE: The aim of this study was to investigate the role of glycan moiety of a C. arizonica extract in the binding of patients' IgE and to identify the carbohydrates possibly involved. METHODS: The reactivity of IgE with C. arizonica extract, before and after periodate-treatment, was evaluated by immunoblotting and ELISA inhibition. The specificity of carbohydrate-reactive IgE was evaluated by ELISA using unrelated glycoproteins with known sugar composition and structure, such as pineapple bromelain, honeybee venom phospholipase A2, and ovalbumin, before and after periodate treatment. RESULTS: When periodate-treated C. arizonica extract was probed after SDS-PAGE and immunoblotting with patients' IgE, no reactivity could be detected. Furthermore, a very poor inhibitory activity of the periodate treated C. arizonica extract as compared with the untreated sample could be observed in the ELISA inhibition experiments performed using C. arizonica extract as antigen. When phospholipase A2 and bromelain were used as antigens in ELISA, they were recognized by patients' IgE, whereas ovalbumin was negative. Treatment of phospholipase A2 and bromelain with periodate completely abolishes the IgE reactivity. CONCLUSION: A large portion of the IgE reactivity of Cupressaceae allergic subjects appears to be associated with sugar moieties of C. arizonica extract which appear to be shared by bromelain and phospholipase A2, thus suggesting that the IgE of patients reacting with such epitopes probably react with beta 1 --> 2 xylose, alpha 1 --> 3 fucose and/or alpha 1 --> 6 fucose. PMID- 10457114 TI - Human immunoglobulin isotype profiles produced in response to antigens recognized by monoclonal antibodies specific to Anisakis simplex. AB - BACKGROUND: Anisakis simplex is a medically important pathogen which not only causes anisakiasis but may provoke allergy reactions, ranging from mild urticaria to anaphylactic shock. OBJECTIVE: To investigate anti-Anisakis isotype profiles in anisakiasis and Anisakis allergy patients. METHODS: Capture ELISA techniques were used to investigate the isotype profiles of antibodies specific for two defined Anisakis simplex antigens, in serum from Japanese patients with confirmed anisakiasis and from Spanish patients with allergy to Anisakis. The antigens were 'UA2R antigens' (two proteins with MW of 48 and 67 kDa, recognized by our monoclonal antibody UA2) and 'UA3R antigens' (two proteins with MW of 139 and 154 kDa, recognized by our monoclonal antibody UA3). RESULTS: Considering IgG, the two most frequent isotypes in the response to the UA2R antigens were IgG1 and IgG2, with IgG4 detected in only one case; in response to the UA3R antigens, by contrast, the two most frequent isotypes were IgG1 and IgG4 (though IgG2 remained reasonably frequent). As regards potential utility for serodiagnosis, 95% of the Japanese anisakiasis patients and 84% of the allergy patients showed detectable IgG1 antibodies to the UA3R antigens. Furthermore, all allergy patients showed IgE antibodies to these antigens. CONCLUSION: Anisakis simplex contains antigens that induce responses which are differentially regulated. Because of their immunogenicity, immunodominance and allergenic nature, we consider that the 139/154-kDa antigens recognized by our MoAb UA3 are good candidates for use in tests for the diagnosis of anisakiasis and of the allergy caused by this parasite. PMID- 10457115 TI - Citrus red mite (Panonychus citri) is the most common sensitizing allergen of asthma and rhinitis in citrus farmers. AB - OBJECTIVE: To evaluate type I hypersensitivity to citrus red mite (Panonychus citri), its prevalence, and relationship to respiratory dysfunction, a cross sectional survey was performed among citrus farmers on Cheju Island, Korea. MATERIALS AND METHODS: Questionnaires, and skin prick test responses to 11 common inhalant allergens and citrus red mite were performed in 181 citrus farmers, and serum-specific IgE antibodies to citrus red mite were measured by ELISA in sera of 123 subjects. To determine airway hyperresponsiveness, methacholine bronchial provocation tests were performed in 55 subjects who complained of recurrent lower respiratory symptoms. RESULTS: The prevalence of asthma-based on presence of asthmatic symptoms on the questionnaire and airway hyperresponsiveness to methacholine, and allergic rhinitis based on presence of nasal symptoms on the questionnaire and positive skin-test response were 12.1% and 19.3%, respectively. The positive rate of skin responses to one or more of 11 common inhalant allergens excluding citrus red mite was 17.1%, and if citrus red mite was included, 25.9% of farmers had positive responses. On skin prick tests, citrus red mite (16.5%) was the most common sensitizing allergen, followed by cockroach (11.0%), Dermatophagoides pteronyssinus (9.9%), and D. farinae (9.3%). Among farmers with asthma and allergic rhinitis, the positive skin responses to citrus red mite were noted in 54.5 and 68.5%, respectively. Serum-specific IgE antibodies to citrus red mite were detected in 45 farmers (36. 5%) of the 123 tested, and there was significant correlation between specific IgE level and weal (A/H ratio) to citrus red mite (r = 0.57, P < 0.001). The prevalence of asthma was higher in subjects with positive skin responses or high serum-specific IgE antibodies to citrus red mite than in those without skin response or serum specific IgE (P < 0.05, respectively). CONCLUSION: Citrus red mite is the most important allergen in citrus farmers with asthma and rhinitis in which causative allergen has not been identified. It should be included in the skin test battery for screening the causative allergen in farmers exposed to citrus red mite. PMID- 10457116 TI - Possible influences of Staphylococcus aureus on atopic dermatitis-- the colonizing features and the effects of staphylococcal enterotoxins. AB - BACKGROUND: Heavy colonization of atopic dermatitis (AD) with Staphylococcus aureus is well documented. This phenomenon suggests that S. aureus in AD lesions influences the disease processes of AD. OBJECTIVE: We describe the importance of the presence of S. aureus and staphylococcal enterotoxins A and B (SEA, SEB) in AD lesions. METHODS: We investigated the colonizing features of S. aureus in AD lesions using electron microscopy, the distribution of SEB in the eczematous skin of AD using immunofluorescence, the effects of SEA and SEB on normal human epidermal keratinocytes in organ culture, and the presence of specific IgE antibodies to SEA and/or SEB in serum of AD patients by enzyme immunoassay. RESULTS: S. aureus in AD lesions colonized on and in the horny layers of the eczematous skin. SEB produced by S. aureus was distributed mainly on the dermal infiltrated cells, especially on eosinophils. SEA and SEB stimulated expression of ICAM-1 and HLA-DR in normal human keratinocytes. More than half of the AD patients in the present study had specific IgE antibodies to SEA and/or SEB in their serum. CONCLUSION: S. aureus and SEs have important roles in the exacerbation and prolongation of AD. PMID- 10457117 TI - Ribonuclease-gold ultrastructural localization of heparin in isolated human lung mast cells stimulated to undergo anaphylactic degranulation and recovery in vitro. AB - BACKGROUND: Human mast cells are a rich and unique source of heparin, which is stored in cytoplasmic secretory granules and accounts for metachromasia, a staining property used to identify mast cells by light microscopy. OBJECTIVE: The sub-cellular locations of heparin in secretory human mast cells and human mast cells recovering from secretion are not known. Acquisition of this knowledge requires ultrastructural imaging of well-preserved cells with a visible probe which binds to heparin. We sought to develop this knowledge regarding human mast cell secretion by using a labelling method for heparin that depends on the well known property of ribonuclease inhibition by heparin. METHODS: Human lung mast cells were isolated, partially purified, either stimulated or not stimulated to secrete with anti-IgE, and recovered 20 min or 6 h later for routine electron microscopy. Histamine secretion was also determined. A previously developed post embedding, enzyme-affinity-gold electron microscopic technique to image ribonucleic acid (RNA) with ribonuclease-gold (R-G), which also binds to the enzyme inhibitor, heparin, was employed to determine the sub-cellular locations of heparin in non-secretory and secretory mast cells as well as in mast cells recovered from short-term cultures after secretion. Specificity controls for the novel use of this method and quantification of granule labelling in these controls were performed. RESULTS: Heparin was labelled by R-G in electron-dense granules within non-secretory human lung mast cells (HLMCs), in electron-dense granules that persisted in secretory HLMCs at the maximum histamine secretion time (20 min), and in electron-dense granules within recovering HLMCs. Specificity controls showed that gold alone did not label HLMCs and that absorption with heparin significantly reduced or abrogated HLMC granule staining with R-G, but that RNA absorption did not. Heparin stores were absent in newly formed, electron-lucent intracytoplasmic degranulation channels in secretory HLMCs. Electron-dense granule matrices in the process of extrusion to the cell exterior still retained heparin at the instant of cellular secretion. Non-granule heparin stores bound R-G in recovering HLMCs. These locations included resolving degranulation channels, as newly emergent granules partitioned and condensed within them, and electron-dense content-containing vesicles and progranules within synthetic mast cells. Ultimately, all known ultrastructural patterns of HLMC granules developed in recovering cells, and each of them contained heparin. CONCLUSION: Heparin was secreted from HLMCs which were stimulated by anti-IgE, and heparin was recovered by a combination of conservative and synthetic mechanisms in HLMCs after a secretory event. PMID- 10457118 TI - Eicosapentaenoic acid inhibits prostaglandin D2 generation by inhibiting cyclo oxygenase-2 in cultured human mast cells. AB - BACKGROUND: Eicosapentaenoic acid (EPA) is catalysed by cyclo-oxygenase (COX), as is arachidonic acid, and is a competitive inhibitor of arachidonate metabolism. OBJECTIVES: We examined the effect of EPA on prostaglandin (PG) D2 generation in the cultured human mast cells with IgE-anti-IgE challenge incubation. METHODS: Cultured human mast cells were incubated with EPA (1 micromol/L) for 20 h, then challenged with anti-IgE incubation after treatment with IgE. At the same time, COX inhibitors were tested to identify COX-1 and COX-2 activity. PGD2 synthetic activity was also assayed in a cell-free homogenate of cultured mast cells with COX inhibitors and EPA. Histamine in the culture medium and in cells was assayed with the HPLC-fluorescent method. PGD2 and PGD3 were assayed with gas chromatography-mass spectrometry and the stable isotope dilution method. RESULTS: Although EPA incubation did not affect histamine release by cultured human mast cells in response to IgE-anti-IgE challenge incubation, it did decrease PGD2 generation by inhibiting the COX-2 pathway. In contrast, in the cell-free homogenate of cultured human mast cells, EPA inhibited both COX-1 and COX-2 activities. CONCLUSION: Pre-incubation with EPA primarily affects the COX-2 pathway in cultured human mast cells and reduces PGD2 generation in response to IgE-anti-IgE challenge incubation. These findings suggest that COX-1 and COX-2 have different substrate flow systems in mast cells. They also suggest that endogenous EPA diet supplementation would reduce PGD2 production and could serve as an anti-inflammatory substrate in human mast cells. PMID- 10457119 TI - Nitric oxide metabolites in induced sputum: a marker of airway inflammation in asthmatic subjects. AB - BACKGROUND AND OBJECTIVE: The role of nitric oxide (NO) needs to be further clarified in allergic inflammation. This study was designed to investigate the relationships between NO metabolites and eosinophil count, eosinophil cationic protein (ECP), interleukin (IL)-5 in induced sputum from asthmatics. METHODS: Hypertonic saline-induced sputum was obtained in 25 asthmatic subjects, among which 13 patients were examined before and after anti-asthmatic medications including steroid preparations. Ten normal subjects were enrolled as controls. Fresh expectorated sputum separated from saliva was treated with equal volume of dithiothreitol 0.1%, cytospinned for cell count, and the supernatant was collected for biochemical assay. NO metabolites were assayed by using modified Griess reaction. ECP was measured by fluoroimmunoassay, and detected IL-5 by a sandwich ELISA. RESULTS: Asthmatic subjects, compared with controls, had significantly higher concentration of NO metabolites (1035.4 +/- 125.3 vs 557.2 +/- 101.5 micromol/L, P < 0.01), higher percentage of eosinophils (25.6 +/- 4.6 vs 1.7 +/- 0.2%, P < 0.01), and higher levels of ECP (1117.8 +/- 213.9 vs 154.6 +/- 47.4 microg/L, P < 0.01) in the induced sputum. IL-5 was detected more frequently in asthmatic subjects than in control subjects (11/25 [44%] vs 1/10 [10%], P < 0.05). According to asthma severity, moderate to severe asthmatic subjects (n = 18) had higher level of NO metabolites (1143.8 +/- 156.3 vs 575.5 +/- 89.5 micromol/L, P < 0. 01), higher levels of ECP and IL-5 (P < 0.01, respectively) in the induced sputum than in those of mild asthmatic subjects (n = 7). NO metabolites, the percentage of eosinophils, the levels of ECP, and IL-5 were reduced following treatment with anti-asthmatic drugs (P < 0.01, respectively). There were significant positive correlations between NO metabolites and percentage of eosinophils or ECP (r = 0. 34, P < 0.05; r = 0.28, P < 0.05). Negative correlations were noted between FEV1, FEV1/FVC and proportion of eosinophils, ECP, or IL-5 levels. CONCLUSION: These findings confirmed that the level of NO metabolites was increased in the tracheobronchial secretion of asthmatic subjects and was paralleled with severity of asthma. Measurement of NO metabolites in induced sputum may be used for monitoring the degree of airway inflammation in asthmatics. PMID- 10457120 TI - A controlled study on the effectiveness of loratadine in combination with flunisolide in the treatment of nonallergic rhinitis with eosinophilia (NARES). AB - BACKGROUND: Nonallergic rhinitis with eosinophilia (NARES), accounting for some 15% of perennial rhinitis, is a nasal disorder whose main features are eosinophil counts in nasal smear higher than 10% and negative IgE tests. The mainstay of treatment is topical corticosteroids. OBJECTIVE: To evaluate the adjunctive effect of loratadine, a non-sedating antihistamine with anti-allergic activity, on nasal symptoms and eosinophil counts in nasal secretions in patients with NARES. METHODS: Thirty patients with NARES were divided in two groups, half receiving flunisolide two 25 microg puffs per nostril morning and night plus loratadine 10 mg u.i.d. and half the same doses of flunisolide plus placebo, according to a double-blind fashion, for 3 weeks. The effectiveness of the treatment in the two groups was evaluated by comparing symptom scores and eosinophil counts, and safety was assessed by comparing the adverse effects. RESULTS: The loratadine treated group had better results both in nasal symptoms, with a decrease in sneezing (P < 0.000001) and rhinorrhoea (P < 0.006), respectively, corresponding with 73.4% and 66.7% with respect to 46.6% and 26.7% in the control group, and in eosinophil counts which decreased by 20% compared with 14.3% in patients treated with placebo and flunisolide. As to safety, only nasal irritation in two patients, one in each group, was reported. CONCLUSION: Loratadine improves the effectiveness of flunisolide in treatment of NARES with no change in safety, and with no sedation. PMID- 10457121 TI - Natural rubber latex allergy. AB - Rubber is an important industrial and consumer product encountered in many household items and medical devices. Whereas the chemical additives used in its manufacture are well recognized as a cause of delayed-type hypersensitivity (allergic contact dermatitis), it is only during the past decade that immediate type allergy to natural rubber latex (NRL) proteins (latex allergy) has emerged as a serious health issue. Frequent, prolonged wearing of NRL gloves, especially amongst health care workers, is a major risk factor for such sensitization. Latex allergy is now an important medical, occupational, medico-legal and financial problem, and it is essential that policies are developed to reduce it. Undiagnosed latex allergy is potentially very serious for patients. This article briefly overviews latex allergy with emphasis on the practical issues of diagnosis and management. PMID- 10457122 TI - Dermatological adverse effects with the antimalarial drug mefloquine: a review of 74 published case reports. AB - Mefloquine is a relatively new antimalarial drug which has been associated with a wide variety of adverse effects, including skin reactions. In order to evaluate the range and frequency of mefloquine's dermatological effects, we searched the scientific literature for published case reports of such effects. We found 74 case reports, published between the years 1983 and 1997. Pruritus and maculopapular rash are the dermatological effects most commonly associated with mefloquine: their approximate frequency is 4-10% for pruritus, and up to 30% for nonspecific maculopapular rash. Adverse effects associated less commonly with mefloquine include urticaria, facial lesions and cutaneous vasculitis. One case of Stevens-Johnson syndrome and one fatal case of toxic epidermal necrolysis occurred. Appropriate primary studies of mefloquine use should be carried out to elucidate the epidemiology and aetiology of dermatological and other adverse effects of the drug. PMID- 10457123 TI - Clinical features and management of 87 patients with pemphigoid gestationis. AB - Pemphigoid gestationis is a rare vesiculo-bullous disorder of pregnancy. In this review we summarize the clinical data on 142 pregnancies in 87 patients complicated by pemphigoid gestationis. Our aim is to provide a comprehensive clinical overview of this disease. PMID- 10457124 TI - Toxic epidermal necrolysis and graft vs. host disease: a clinical spectrum but a diagnostic dilemma. AB - We describe a 53-year-old man who developed partial and full thickness skin loss associated with pyrexia, diarrhoea, liver, renal and bone marrow failure, during treatment for an aggressive B cell lymphoblastic lymphoma. The clinical features and histology were compatible with both toxic epidermal necrolysis and graft vs. host disease, causing a diagnostic and therapeutic dilemma. We discuss the possibility that methotrexate was the causative drug, with review of its cutaneous side-effects. Histologically our patient demonstrated the sparse dermal infiltrate with full thickness epidermal necrosis typical of toxic epidermal necrolysis and graft vs. host disease. We discuss this finding with respect to the pathogenesis of toxic epidermal necrolysis. PMID- 10457125 TI - Bullous pemphigoid developing during systemic therapy with chloroquine. AB - Bullous pemphigoid has been reported to be induced or precipitated by systemic therapy with several drugs, including penicillamine, captopril, frusemide and ampicillin. We report an African male patient with sarcoidosis who was prescribed chloroquine for progressive dyspnoea. After 3 months he developed generalized pruritus which evolved into a widespread bullous eruption with acral targetoid lesions resembling erythema multiforme. The histological and immunofluorescence findings were diagnostic of bullous pemphigoid. The atypical clinical features of this case resemble the phenotype that has been noted in previous reports of drug induced bullous pemphigoid. PMID- 10457126 TI - Lichen planus associated with milia. AB - The formation of milia is well recognized in both bullous and inflammatory dermatoses. There are several reports of milia developing in a rare variant of lichen planus pilaris known as lichen planus follicularis tumidus (LPFT), but the association of milia with other types of lichen planus (LP) has not been documented in the literature. We report five patients who developed milia during the course of either drug-induced or idiopathic LP and one in whom milia developed in a lichenoid tattoo reaction. Milia were noted to occur transiently during the resolving phase of LP. Most cases were severe enough to warrant treatment with systemic steroids. The association of milia with LP is not restricted to the rare clinical variant LPFT. We speculate that a severe lichenoid reaction with basal layer degeneration may precipitate the formation of milia in some cases of LP. PMID- 10457127 TI - Chronic lymphoedema and angiosarcoma. AB - Angiosarcoma has frequently been described arising within chronic lymphoedema of the upper limb following mastectomy and radiotherapy for carcinoma of the breast. We report a case of angiosarcoma arising in a lymphoedematous leg that had been subjected to radiotherapy 20 years previously for Hodgkin's disease. The diagnosis was expedited once the patient noticed the development of bleeding nodules. Prognosis of angiosarcoma is poor with treatment options being wide excision surgery, palliative radiotherapy or chemotherapy. Unusual bruised areas or bleeding nodules developing within chronic lymphoedematous limbs should be biopsied to exclude the diagnosis. PMID- 10457128 TI - Polymorphic light eruption occurring in common variable hypogammaglobulinaemia, and resolving with intravenous immunoglobulin therapy. AB - A 55-year-old woman with a past history of lower respiratory tract infections presented with a photosensitive eruption. Polymorphic light eruption (PLE) was diagnosed on the basis of the temporal relationship to sun exposure and the diagnosis was supported by positive monochromator irradiation tests in the ultraviolet A wavelength spectrum. Investigation of the patient's immune status identified low levels of all immunoglobulin (Ig) subtypes consistent with common variable hypogammaglobulinaemia. Intravenous Ig replacement therapy, instituted to minimize risks from bacterial infections, was commenced and over the ensuing months resulted in a complete resolution of the PLE. PLE is considered to represent a type IV hypersensitivity reaction directed against a cutaneous autoantigen induced by exposure to ultraviolet light. In PLE, nonspecific immunomodulatory mechanisms of intravenous Ig may be active, such as a reduction in the synthesis of cytokines and a blockage of the IgG Fc receptors on macrophages. PMID- 10457129 TI - Dermatomyositis with elevated serum hyaluronate. AB - We report two Japanese patients with dermatomyositis whose serum hyaluronate levels were correlated with their disease activities. The diagnosis of one patient was primary idiopathic dermatomyositis, and that of the second patient was dermatomyositis associated with neoplasia. The serum hyaluronate level of the former patient was decreased by systemic corticosteroid therapy, and that of the latter patient decreased following the surgical resection of mammary carcinoma and subsequent chemotherapy. The disease activity of each patient was greatly improved after these therapies. The longitudinal measurement of serum hyaluronate concentration may be useful to estimate the disease activity of patients with dermatomyositis whose initial serum levels of hyaluronate are elevated. PMID- 10457130 TI - Vulvar myiasis due to Wohlfahrtia magnifica. AB - Myiasis is a condition resulting from the invasion of tissues or organs of man or animals by the larvae of dipterous flies. The distribution of myiasis is worldwide, with more cases being reported from tropical, subtropical, and warm temperate areas. The various forms of myiasis may be classified from an entomological or a clinical point of view. This report describes a rare case of vulvar myiasis due to Wohlfahrtia magnifica in an otherwise healthy 20-year-old Iranian female. To our knowledge, this is the first report of cutaneous myiasis of the vulva due to W. magnifica from Iran and the Middle East region. PMID- 10457131 TI - Multiple basal cell carcinomas in a patient with acute myeloid leukaemia and chronic lymphocytic leukaemia. AB - Skin cancer is a well-recognized risk of prolonged immunosuppression, for example, following renal transplantation. These tumours contrast with idiopathic lesions in that squamous cell, rather than basal cell carcinomas usually predominate. We report a Caucasian female who developed multiple basal cell carcinomas following protracted cytotoxic therapy for acute myeloid leukaemia and subsequently chronic lymphocytic leukaemia. No other clinical risk factors nor relevant polymorphisms of genes encoding for detoxifying enzymes were identified. Immune suppression is a well-recognized cause of multiple skin tumours, the most striking increase usually being of squamous cell carcinomas. We believe this woman is representative of a subgroup of immunosuppressed patients who, for as yet poorly understood reasons, have a predisposition to basal cell, rather than squamous cell carcinoma accrual. PMID- 10457132 TI - Childhood dermatitis herpetiformis: an unusual presentation. AB - Childhood dermatitis herpetiformis (DH) is rare. The true prevalence and incidence of this condition are unknown. We report a 7-year-old boy presenting with nonpruritic inflammatory papules on the buttocks and extensor surfaces, clinico-pathologically consistent with Sweet's neutrophilic dermatosis. Immunofluorescence studies showed IgA deposits in the dermal papillae consistent with DH. Remission was achieved with a gluten-free diet and dapsone. Childhood DH may present different clinical signs to the adult form and misdiagnosis can occur if immunofluorescence is not requested on skin biopsy. PMID- 10457133 TI - The role of twin studies in the genetics of skin diseases. AB - With advances in molecular biology and statistical techniques, gene discovery is moving at a fast pace. Most skin diseases have a complex mode of inheritance and the twin design is the ideal model to dissect the genetic architecture. Use of the twin pairings in a variety of ways for associations and linkage studies can enhance gene discovery. This review covers the advantages and limitations of the twin method. PMID- 10457134 TI - Which T cells cause psoriasis? AB - Today, T cells appear to be the main protagonists in the pathogenesis of psoriasis vulgaris. This article summarizes how T cells might contribute to the generation of psoriatic skin lesions. It discusses the preferential T cell receptor usage and the putative mode of T cell activation in psoriatic skin lesions, and how streptococcal throat infections could be involved in disease manifestations. The results are integrated into a pathogenetic concept which considers psoriasis as a T-cell mediated autoimmune disorder. PMID- 10457135 TI - Mutations in the translation initiation codon of the protoporphyrinogen oxidase gene underlie variegate porphyria. AB - Variegate porphyria (VP), one of the acute hepatic porphyrias, is characterized by a reduced catalytic activity of protoporphyrinogen oxidase (PPO), the penultimate enzyme in the porphyrin-haem biosynthetic pathway. VP has been linked to the PPO gene on chromosome 1q22-23, and several mutations underlying this disorder have been described recently. In this study, we identified two different missense mutations in the translation initiation codon of the PPO gene in two unrelated patients with VP. Mutation analysis was carried out using PCR, heteroduplex analysis, automated sequencing, and restriction enzyme digestion. In the first patient, the results revealed an A-to-T transversion (ATG --> TTG), resulting in the substitution of methionine by leucine (M1L). The mutation detected in the second patient was a T-to-C transition (ATG --> ACG), leading to the conversion of methionine to threonine (M1T). These mutations abolish the initiation of translation at the normal site, and consequently, translation of an abnormal messenger RNA (mRNA) would result in the synthesis of a truncated PPO protein lacking the amino terminus. PMID- 10457136 TI - IgA basement membrane zone autoantibodies in bullous pemphigoid detect epidermal antigens of 270-280 kDa, 230 kDa, and 180 kDa molecular weight by immunoblotting. AB - Bullous pemphigoid (BP) is an acquired subepidermal blistering disease characterized by circulating IgG autoantibodies binding to the 230 and 180 kDa hemidesmosomal proteins. Associated basement membrane zone (BMZ) autoantibodies of the IgA class have been reported in few BP patients. The incidence and clinical relevance of these IgA antibodies, as well as their target antigens are unknown. Sera of 26 patients with BP were analysed for circulating IgG- and IgA anti-BMZ autoantibodies by indirect immunofluorescence on salt-split human skin. All of the patients had circulating IgG autoantibodies and, in addition, nine (35%) also had circulating anti-BMZ IgA antibodies, that bound to the epidermal side of salt-split skin. By immunoblotting, IgA antibodies in seven of nine sera recognized either the 180 kDa, the 230 kDa, or both BP antigens. Moreover, IgA anti-BMZ antibodies in seven sera also detected an epidermal protein of 270-280 kDa. IgA antibodies did not identify specific bands on immunoblots of dermal extracts. There was no clinical difference between BP patients with or without circulating anti-BMZ-IgA. PMID- 10457137 TI - Passive sustainable hydration of the stratum corneum following surfactant challenge. AB - Surfactants exhibit a protein-denaturating potency which is responsible for water uptake and swelling of the stratum corneum. There is also an increased transepidermal water loss related to the alteration of the barrier function. In the present prospective study, we evaluated sodium lauryl sulphate (SLS) dependent changes in electrometric properties of skin. Single and iterative SLS challenges were performed. Measurements were made using a NOVA DPM on continuous mode for 5 min in order to assess both the baseline surface hydration and the rate of changes during probe occlusion. The present noninvasive instrumental evaluation of surfactant irritancy shows high sensitivity in each subject, but suffers from large inter-individual variability in the skin response following single and iterative sodium lauryl sulphate (SLS) challenges. The baseline surface hydration, that is almost unaffected 1 h after removing SLS patches, significantly decreases after a lag time of 24 h. The rate of water accumulation in the stratum corneum during the probe occlusion is significantly increased by SLS and is proportional to the SLS concentration and the number of iterative patches. Single water- or SLS-patch-tests fail to predict the skin response to cumulative SLS challenges and inter-individual variability is large for all electrometric variables. PMID- 10457138 TI - Microscopic Nikolsky's sign. AB - Nikolsky's sign is a useful clinical sign in patients with active pemphigus. It does not however, indicate the level of split in the skin, and is seldom present in partially-treated patients. The purpose of this study was two fold: (i) to document the microscopic or subclinical counterpart of clinical Nikolsky's sign; and (ii) to improve the diagnostic yield of routine histopathology in pemphigus patients in whom the lesions are infected/old and are therefore not useful for histopathology, and when intact blisters are not present; this is particularly useful for institutions in which immunofluorescence facilities are not available. Pemphigus patients were allocated to one of two groups. Group A patients (n = 23) were subjected to manual tangential pressure over the perilesional skin before a biopsy specimen was taken from that site; group B patients (n = 14) were subjected to a biopsy without the tangential pressure technique. Group C consisted of 37 healthy volunteers who were subjected to the tangential pressure technique before a biopsy. Histopathological changes of pemphigus vulgaris or foliaceus were present in 73.9% patients in group A, 28.6% in group B and none in the control group C. Tangential pressure as described below can produce microscopic changes in the epidermis which are diagnostic of pemphigus. These changes are produced at the suprabasal level in pemphigus vulgaris and intraepidermally in pemphigus foliaceus. This technique is of value in those parts of the world where immunofluorescence facilities are not readily available. PMID- 10457139 TI - A long-term time course of colorimetric evaluation of ultraviolet light-induced skin reactions. AB - Many attempts have been made to quantify ultraviolet (UV) radiation-induced erythema and pigmentation. However, most of these studies were concerned with the early changes of reactions and neglected events occurring in later stages. The long-term course of skin colour changes in pigmented skin, induced by broad band UVA and UVB radiation, was evaluated in 30 Korean male volunteers by means of a tri-stimulus colorimeter for 10 weeks. The L*a*b* system recommended by the Commission International de l'Eclairage was used to measure skin colour. The L* value (luminance) gives the relative lightness ranging from total black to total white. The a* value represents the balance between red and green and the b* value the balance between yellow and blue. The mean individual typology angle of our subjects was 47.3 degrees, indicating 'light' group of constitutional skin colour category. One day after UV exposure, the L* and b* values decreased significantly, following the colour direction of persistent pigment darkening. They then changed in opposite directions persistently until week 1, when maximum tanning was obtained. Then, a shift toward the original values was observed parallel to the constitutive melanization axis. The a* index showed a significant increase toward the mean colour of haemoglobin on day 1. It returned to its original value following the pathway of constitutive melanization axis. This promising quantitative method may enable objective measurement of dermatophysiologic changes to be made, and allow evaluation of the efficacy of therapeutic modalities on skin disorders without the inherent errors associated with subjective judgement. Our results would provide standard data for long-term UV-induced skin erythema and pigmentation. PMID- 10457140 TI - The role of interleukins 1, 6 and 8 as lymphocyte attractants in the photodermatoses polymorphic light eruption and chronic actinic dermatitis. AB - The two photodermatoses, polymorphic light eruption (PLE) and chronic actinic dermatitis (CAD), are characterized by lymphocyte-rich inflammatory infiltrates, the pathogeneses of which are not fully understood. We have therefore studied suction blister fluid (SBF) samples from patients with these conditions before and at two time points after the induction of experimental lesions by means of a solar simulator; this SBF was then tested for the presence of selected cytokines known to induce peripheral blood lymphocyte (PBL) migration in vitro. A specific EL-4 NOB-1 bioassay was used to detect interleukin (IL)-1 activity, which has already been noted in normal skin and this was found in pre-irradiation control samples as well as 1-3 h and 24 h post-irradiation in both patient groups, but at levels not significantly different from those of controls. Use of a B9 cell proliferation assay showed no detectable IL-6-like activity pre-irradiation, but there was substantial activity in samples at both post-irradiation time points in both patient groups. Further, in other experiments, retained SBF samples were tested in an in vitro PBL migration assay in the presence and absence of neutralizing antibodies against IL-1 alpha, IL-1 beta, IL-6 and IL-8; considerable PBL attractant activity was noted in the pre-irradiation SBF from both patient groups; a finding consistent with previous reports of such activity in samples from normal skin, and at least in CAD patients, a proportion of this activity appeared to be due to IL-1, pre-incubation of SBF with neutralizing antibodies against IL-1 alpha and IL-1 beta reducing the effect significantly. Substantial PBL attractant activity was present also in the SBF from 1-3 h and 24 h post-irradiation samples in both patient groups and again, IL-1 neutralizing antibodies reduced this in the 1-3 h and 24 h CAD samples. In addition, neutralizing antibodies against IL-6 and IL-8 reduced the activity in the 24 h PLE samples significantly and although not fully conclusive in the case of IL-1, these data suggest that IL-6, IL-8 and possibly IL-1 may be involved in the induction of PBL infiltrates, and perhaps other events, in both PLE and CAD. PMID- 10457141 TI - Herpes zoster in Japanese patients with systemic lupus erythematosus. AB - We examined retrospectively the incidence of herpes zoster among 58 Japanese patients with systemic lupus erythematosus (SLE), 42 patients with systemic sclerosis (SSc) and 21 patients with autoimmune bullous diseases (AIBD). The incidence of herpes zoster was significantly higher in SLE (46.6%) than in SSc (9.5%) or AIBD (4.7%) (P < 0.001). The rate of positive delayed-type skin test with varicella zoster virus antigens was significantly lower in SLE than in SSc [4/16 (25.0%) vs. 11/12 (91.7%), P < 0.001]. In addition, the positive rate of skin test with purified protein derivatives of tuberculin was also low in SLE patients (3/16, 18.8%). These results reconfirm the general impairment in cellular immunity in SLE. PMID- 10457142 TI - Hydroxyurea-induced skin disease mimicking the baboon syndrome. PMID- 10457143 TI - Multiple squamous cell carcinomas following photochemotherapy for atopic eczema. PMID- 10457144 TI - Bowen's disease--a retrospective review of clinical management. PMID- 10457145 TI - Activation of complement in psoriasis. PMID- 10457146 TI - The complement system: a pathway linking host defence and adipocyte biology. PMID- 10457147 TI - Future for folates in cardiovascular disease. PMID- 10457148 TI - Oral folate enhances endothelial function in hyperhomocysteinaemic subjects. AB - BACKGROUND: Elevated plasma homocysteine (Hcy) is a risk factor for vascular disease. A postulated mechanism is vascular endothelial damage by homocysteine. Hcy levels are inversely related to blood concentrations of folate and can be lowered by folate supplements. The effect of oral folic acid on endothelial function was investigated in healthy adults with mild hyperhomocysteinaemia. PATIENTS AND METHODS: Eighteen healthy subjects (Hcy > 13 micromol L-1 at entry), from a screening population of 890 volunteers, were entered into a randomised double-blind placebo-controlled crossover study of oral folic acid (5 mg daily for six weeks) with a six week interval between treatments. Flow-mediated (endothelium-dependent) and (endothelial-independent) glyceryl trinitrate (GTN) mediated brachial artery dilatation were measured by high resolution wall tracking. RESULTS: Folate supplementation enhanced endothelium-dependent responses (+0.08 +/- 0.05 vs. +0.04 +/- 0.04 mm, P = 0.015) but endothelium independent responses (GTN) were unchanged. Folate reduced Hcy (8.7 +/- 2.5 vs. 12.1 +/- 3.6 micromol L-1). CONCLUSION: High dose folic acid supplementation enhances endothelium-dependent vascular function and lowers plasma Hcy. This provides preliminary evidence that folate may have beneficial cardiovascular effects in adults with mild hyperhomocysteinaemia. PMID- 10457149 TI - Insulin decreases plasma cholesteryl ester transfer but not cholesterol esterification in healthy subjects as well as in normotriglyceridaemic patients with type 2 diabetes. AB - BACKGROUND: Plasma cholesterol esterification (EST) and subsequent cholesteryl ester transfer (CET) from high-density lipoproteins (HDLs) towards apolipoprotein (apo) B-containing lipoproteins are key steps in HDL metabolism. MATERIALS AND METHODS: The effects of exogenous hyperinsulinaemia on plasma CET and EST, measured with isotope methods, were evaluated in 10 male normotriglyceridaemic (plasma triglycerides <2.0 mmol L-1) patients with type 2 diabetes and 10 individually matched healthy subjects during a two-step hyperinsulinaemic euglycaemic clamp over 6-7 h. RESULTS: No between-group differences in baseline plasma lipid parameters were observed, but the HDL cholesteryl ester content was lower (P < 0.02) and the HDL triglyceride content was higher (P < 0.05) in diabetic patients. Baseline CET and EST were similar in the groups. In both groups, hyperinsulinaemia decreased plasma triglycerides (P < 0.01) and the HDL triglyceride content (P < 0.01) compared with saline infusion in healthy subjects, whereas the HDL cholesteryl ester content increased (P < 0.05 vs. saline infusion) in diabetic patients. CET was similarly decreased by hyperinsulinaemia in both groups (P < 0.01 vs. saline infusion). In contrast, the change in EST in either group was not different from that during saline administration. In the combined group, baseline CET was positively correlated with plasma triglycerides (Rs = 0.68, P < 0.01). The HDL cholesteryl ester content was negatively (Rs = -0.48, P < 0.05) and the HDL triglyceride content was positively (Rs = 0.64, P < 0.01) correlated with CET. CONCLUSION: Insulin infusion decreases plasma CET in conjunction with a fall in triglycerides but does not decrease cholesterol esterification in healthy and type 2 diabetic subjects, indicating that acute hyperinsulinaemia has a different effect on these processes involved in HDL metabolism. Despite unaltered fasting plasma CET, HDL core lipid composition was abnormal in diabetic patients, suggesting that additional mechanisms may contribute to changes in HDL metabolism in diabetes mellitus. PMID- 10457150 TI - Expression pattern of tumour necrosis factor receptors in subcutaneous and omental human adipose tissue: role of obesity and non-insulin-dependent diabetes mellitus. AB - BACKGROUND: Tumour necrosis factor alpha (TNF) mRNA expression has been reported to be up-regulated in adipose tissue from several rodent models of obesity and diabetes and from obese humans. This elevated expression has been assumed to be associated with the development of insulin resistance. However, the biological signal of TNF may be influenced by the expression of the two TNF receptors: the p60 TNF receptor, TNFR60, and the p80 TNF receptor, TNFR80. DESIGN: The aim of this study was to investigate the mRNA expression pattern of the two TNF receptors and their ligand in two adipose tissue depots of glucose-tolerant obese women [n = 18, body mass index (BMI) 48.2 +/- 8.4 kg m-2], obese women with impaired glucose tolerance or overt non-insulin-dependent diabetes mellitus (NIDDM) (n = 10, BMI 49.1 +/- 11.6 kg m-2) and healthy non-obese control subjects (n = 12, BMI 25.8 +/- 2.7 kg m-2). RNA expression was assessed by a semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) technique. RESULTS: The mean mRNA levels of both TNF receptors were two- to threefold higher in adipose tissue samples from the obese than from the non-obese women (P < 0.01 for each). Although TNFR60 mRNA did not vary within each obese group, there was a wide variation in the levels of TNFR80 mRNA and of TNF mRNA. A comparison of the expression levels between the subcutaneous abdominal and the omental adipose tissue depots showed significantly higher expression in the former. The TNFR60 expression level was positively correlated with BMI and fat cell size, whereas TNFR80 and TNF mRNA levels showed positive associations with serum insulin and triglyceride concentrations. No significant differences in the expression levels were observed between obese individuals with and without impaired glucose tolerance/NIDDM. CONCLUSION: These results indicate that severe obesity in women is characterized by increased amounts of the two TNF receptor mRNAs. The role of this up-regulation for the development of obesity-associated insulin resistance remains to be elucidated. PMID- 10457151 TI - Plasma acylation stimulating protein, adipsin and lipids in non-obese and obese populations. AB - BACKGROUND: Acylation stimulating protein (ASP) is a potent stimulator of TG synthesis in human adipocytes. DESIGN: In the present study, we have analysed plasma ASP and adipsin levels and their relationships to plasma lipids in non obese and obese groups. RESULTS: The results show that the frequency distribution of ASP is skewed but that of adipsin is normal in both groups. In the non-obese population, the mean levels of plasma ASP and adipsin were 20.2 nmol L-1 (median) and 66.6 +/- 19 nmol L-1 (mean) respectively. No difference was observed between men and women for each of the parameters. In the obese population, the median plasma ASP was increased by 246% (69.9 nmol L-1) and adipsin by 31% (87.0 +/- 22.7 nmol L-1) above that of the control group. Although the levels for men and women were not statistically different for adipsin, the median ASP plasma concentration was 1.9-fold higher in obese women than in obese men (71.8 nmol L-1 vs. 37.6 nmol L-1, P < 0.05). Best subset regression analysis provided a model with variables that best predict plasma ASP [r2 = 0.160, P < 0.008 for body mass index (BMI), P < 0.05 for triacylglycerol (TG), P < 0.03 for free fatty acid (FFA)] and plasma adipsin (r2 = 0.057, P < 0.017 for BMI) in a non-obese population. In obese subjects, the model was different for plasma ASP (P = NS for any of the variables) and plasma adipsin (r2 = 0.356, P < 0.008 for FFA, P < 0.0002 for BMI, P < 0.02 for age). There was no correlation between ASP and adipsin in either the non-obese or the obese group. CONCLUSION: The present data suggest involvement of the ASP/adipsin pathway in the pathogenesis of obesity. PMID- 10457152 TI - High dose insulin does not increase glucose transfer across the blood-brain barrier in humans: a re-evaluation. AB - BACKGROUND: This study re-evaluates previously published data on blood-brain barrier transfer coefficients in humans exposed to high insulin levels. DESIGN: In this study of seven volunteers, global blood-brain barrier permeability to glucose and phenylalanine was measured by means of the intracarotid double indicator method before, during, and after an insulin-glucose clamp. Data were reanalyzed by means of a mathematical model that takes capillary heterogeneity and labelled glucose backflux from the brain into account. RESULTS: The permeability-surface area product (PS) for glucose transport from the blood into the brain, PS1, was 0.145 (0.102-0.211) (median and quartiles), 0.146 (0.113 0.259), and 0.157 (0.133-0.181) ml g-1 min-1 before, during, and after insulin challenge, respectively. In six of the subjects, PS for transport from brain to blood over the brain glucose distribution volume, PS2/Ve decreased under hyperinsulinemia, from a baseline value of 6.56 (3.0-14.9) to 3.86 (1.41-5.32), and restored to a value of 3.8 (2.8-12.1) min-1 after insulin challenge. This decrease in PS2/Ve is probably due to an increase in the brain glucose distribution volume induced by an insulin induced increased intracellular glucose uptake during the experiment. For phenylalanine (n = 5), PS1 was unchanged before, during, and after insulin challenge. In hyperinsulinemia, PS3/Ve for phenylalanine decreased in all subjects. CONCLUSION: We conclude that acutely elevated high plasma insulin levels in humans does not alter the brain uptake of glucose or phenylalanine from the blood. It seems, however, that high plasma insulin levels induce an increase in the movement of D-glucose and L phenylalanine from the brain interstitial fluid into the intracellular compartment. PMID- 10457153 TI - Increased luminal nitric oxide concentrations in the small intestine of patients with coeliac disease. AB - BACKGROUND: Inflammation is known to be associated with enhanced nitric oxide production. A role for nitric oxide in coeliac disease has been suggested because of increased expression of the inducible isoform of nitric oxide synthase in the small intestine of patients with untreated coeliac disease. DESIGN: During small bowel endoscopy in 11 control subjects, 10 patients with untreated coeliac disease and seven patients with treated coeliac disease, gas was aspirated from different parts of the upper gastrointestinal tract and immediately analysed using a chemiluminescence technique. Luminal nitric oxide concentrations were also quantified in 13 control subjects who had undergone colonoscopy. RESULTS: Jejunal luminal nitric oxide concentrations were more than 20 times higher in patients with coeliac disease than in normal control subjects (mean 755 +/- 173 ppb, range 215-1690 ppb, vs. mean 31 +/- 9 ppb, range 1-83 ppb, P < 0.001). Jejunal luminal nitric oxide levels in patients with treated coeliac disease (mean 54 +/- 18 ppb, range 3-126 ppb) did not differ from those of control subjects. CONCLUSIONS: This study shows that intraluminal jejunal nitric oxide concentrations are significantly increased in patients with untreated active coeliac disease. PMID- 10457154 TI - A novel visual immunoassay for coeliac disease screening. AB - BACKGROUND: To date, the most commonly accepted techniques for the screening of coeliac disease are indirect immunofluorescence and enzyme-linked immunosorbent assay (ELISA), which reveal antiendomysium and antigliadin antibodies respectively. We report the use of a simple visual system for coeliac disease screening based on the use of Staphylococcus aureus protein A, which binds to both IgG and IgA, thus avoiding the need for two parallel immunoassays. MATERIALS AND METHODS: Opaque polystyrene microwell strips coated with a wheat gliadin extract were incubated with sera followed by incubation with protein A-colloidal gold conjugate. The resulting colour was compared with that of positive and negative control sera. The procedure took less than an hour. RESULTS: One hundred and forty-five biopsy-proven sera, 94 from active coeliac patients and 51 from non-coeliac patients with diverse gastrointestinal pathologies or diabetes mellitus, were assayed. Ninety of the 94 sera from the active coeliac patients were positive, whereas only 3 of the 51 non-coeliac control subjects were positive. The technique has a sensitivity of 95.7% and a specificity of 94.1%. CONCLUSIONS: The sensitivity and specificity of the visual system are greater than those of most ELISA systems and are similar to those observed with IgA antiendomysium antibodies when tested in the same population. Moreover, it is inexpensive, quick, simple to perform and easy to interpret, i.e. it requires no qualified personnel. It is for these features, together with the outstanding sensitivity and specificity, that we propose this immunoassay as a new test for reliable coeliac disease screening. PMID- 10457155 TI - Induction of myocardial nitric oxide synthase by Coxsackie B3 virus in mice. AB - BACKGROUND: Inducible nitric oxide synthase (iNOS) expression is regulated by cytokines. This study investigated whether Coxsackie group B virus (CVB) myocarditis resulted in an environment suitable for induction of NOS in the murine heart. MATERIALS AND METHODS: Myocardium was removed from mice infected with CVB3 and from controls. Histology, reverse transcriptase polymerase reaction (RT-PCR) for murine iNOS, NOS enzyme activity and immunohistochemistry were assessed. RESULTS: Histology revealed severe myocarditis 7 days after infection with CVB3 but not in controls. RT-PCR using primers for murine iNOS detected iNOS mRNA in infected mice but not in controls. Calcium-independent NOS activity increased by day 5 after infection with a peak at day 7. Calcium-dependent NOS activity was present throughout, with a trend to lower levels during peak calcium independent activity. Immunohistochemistry revealed iNOS to be localized to inflammatory cells rather than to myocytes. CONCLUSION: This study demonstrates the development of calcium-independent NOS activity and de novo gene transcription for iNOS in the murine myocardium in response to CVB3 infection. The nitric oxide produced at such high output may act at times as part of the immune defence as an antiviral agent and may be toxic to host tissue. PMID- 10457156 TI - Microcirculation in venous ulcers and the surrounding skin: findings with capillary microscopy and a laser Doppler imager. AB - BACKGROUND: To obtain more information about microcirculatory disturbances in venous ulcers, we studied their laser Doppler flux images and capillary densities. MATERIALS AND METHODS: On photographs of venous ulcers and the adjacent skin of 17 patients, four regions of interest were marked: one non granulation tissue area (NGTA) within the venous ulcer, one granulation tissue area (GTA) within the ulcer, one adjacent skin area (ASA) and one distant skin area (DSA). Within these four regions the average laser Doppler area fluxes and capillary densities were determined for each patient using a laser Doppler imager and capillary microscopy respectively. RESULTS: The laser Doppler area flux (mean +/- SD in AU) was significantly lower in NGTA (1.39 +/- 1.12) than in GTA (4.06 +/- 1. 52) or DSA (3.86 +/- 1.54) (P < 0.00001). In addition, the ASA flux (1.95 +/- 1.39) was significantly lower than the GTA or DSA flux (P < 0.0001). Capillary density (capillaries per mm2) in NGTA (0.82 +/- 1.98) was significantly lower than that in GTA (6.00 +/- 2.55), ASA (13.88 +/- 5.16) or DSA (10.29 +/- 3.41) (P < 0.0001). In addition, the capillary density of ASA was significantly higher than that of GTA or DSA (P < 0.05). CONCLUSION: The four areas showed the following characteristics: NGTA, low laser Doppler area flux and lowest capillary density (possible sign of ulcer area without healing tendency); GTA, high laser Doppler area flux and second lowest capillary density (possible sign of wound healing); ASA, low laser Doppler area flux and highest capillary density (possible sign of healing process nearly completed; scar); DSA, high laser Doppler area flux and second highest number of capillaries (sign of microcirculation of chronic venous disorder). PMID- 10457157 TI - Pneumocystis carinii major surface glycoprotein induces interleukin-8 and monocyte chemoattractant protein-1 release from a human alveolar epithelial cell line. AB - BACKGROUND: The major surface glycoprotein (MSG) is an abundant, immunogenic glycoprotein located on the surface of Pneumocystis carinii. Little is known about the proinflammatory effects of MSG. DESIGN: We have investigated the effect of human MSG on the secretion of the chemokines interleukin 8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) from an alveolar epithelial cell line (A549). RESULTS: Incubation of A549 cells with MSG in concentrations from 0.4 to 10 microg mL-1 for 24 h caused dose-dependent increases in IL-8 release (3.4-fold above control, P < 0.01). Time course experiments showed increases in IL-8 release at 4 h, 8 h and 24 h compared with control cultures (all P < 0.01). There was a minor (13%) dose- and time-related increase in MCP-1 release at 24 h (P = 0.02). Co-incubation of MSG with mannan or beta-glucan decreased IL-8 release by 48% and 42% respectively, suggesting that MSG stimulates A549 cells in part through carbohydrate moieties. Dexamethasone significantly inhibited MSG-induced IL-8 release in concentrations of 10-6-10-8 mol L-1 compared with control experiments (P < 0.01). Ribonuclease protection assays for steady-state IL-8 mRNA showed that increases in response to MSG stimulation occurred by 4 h and persisted throughout 8 h of stimulation. CONCLUSION: These findings suggest that MSG can alter alveolar epithelial cytokine release and may be capable of modulating the local inflammatory response in this manner. PMID- 10457158 TI - Laureate ESCI award for excellence in clinical science 1999. Cytokines and the human immunodeficiency virus: from bench to bedside. European Society for Clinical Investigation. AB - Replication of the human immunodeficiency virus (HIV), the causative agent of the acquired immunodeficiency syndrome (AIDS), is under the control of both viral and host factors. Among the latter, the regulatory network of cytokines has been shown to affect virtually every step of the virus life cycle, from cell entry to budding of new progeny virions. Proinflammatory cytokines, such as tumour necrosis factor alpha, can either trigger or potentiate HIV expression via activation of the cellular transcription factor NF-kappaB. Other molecules, including interleukin 6 (IL-6) and the interferons, can up-regulate HIV expression by acting predominantly at post-transcriptional and/or post translational levels. Anti-inflammatory cytokines, including transforming growth factor beta, IL-4 and IL-10, counteract these effects but can also potentiate viral replication under different experimental conditions. Chemotactic cytokines (chemokines) have recently entered the arena of host factors controlling viral spreading as potent inhibitors competing with the virus for cell-surface 7 transmembrane domain receptors also acting, together with CD4, as entry co receptors for HIV. The cytokine network is constitutively activated in most HIV infected individuals, as demonstrated by recent analysis of intracellular signalling molecules such as the Janus kinase/signal transducer and activator of transcription pathway. Finally, cytokines have already shown their potential use as pharmacological agents able to restore at least some of the compromised immune functions in infected individuals, as exemplified by the potent enhancing effect of IL-2 on the number of circulating CD4+ T lymphocytes. PMID- 10457159 TI - Acute reorganization of the forepaw representation in the rat SI cortex after focal cortical injury: neuroprotective effects of piracetam treatment. AB - Immediate postlesion reorganization of the somatosensory cortical representation was examined in adult rats. Response properties of small clusters of neurons were recorded in the area of the primary somatosensory cortex (SI) devoted to the contralateral forepaw representation. Electrophysiological maps were elaborated on the basis of the sensory 'submodality' (cutaneous or noncutaneous) and the location of the peripheral receptive fields (RFs) of layer IV neurons. Recordings were made prior to, and from 1 to 12 h after, induction of a focal neurovascular lesion to the SI cortex that initially destroyed a part (8.5%) of the cutaneous representation. Moreover, the influence of an anti-ischaemic substance (piracetam) on lesion-induced changes was analysed. The main observations were: (i) a gradual outward expansion of the area of the functional lesion, which was smaller in the piracetam-treated (PT) rats than in the control, placebo-treated (PL) rats; (ii) a substantial remodelling of the spared representational zones, both in cortical sectors adjoining the site of injury and those remote from the site; (iii) a significant postlesion increase in the size of cutaneous RFs in the PT rats, but not in the PL rats; (iv) a better preservation of RF submodality and topographic organization in the PT maps than in the PL maps; and (v) a decrease in neuronal responsiveness to cutaneous stimulation which was less pronounced in the PT than in the PL rats. Our results can be ascribed to a rapid change in the balance of excitatory and inhibitory connections which leads to unmasking of subthreshold inputs converging onto cortical neurons. Our findings also indicate that acute piracetam treatment exerts a protective function on the physiological response properties of cortical neurons after focal injury. PMID- 10457160 TI - Post-translational modifications of ICA512, a receptor tyrosine phosphatase-like protein of secretory granules. AB - The autoantigen of type I diabetes ICA512 is a receptor tyrosine phosphatase-like protein enriched in the secretory granule membranes of neurons and peptide secreting endocrine cells. While the function of ICA512 remains unknown, it is thought to link regulated neuropeptide and peptide hormone secretion with signal transduction pathways involving tyrosine phosphorylation/dephosphorylation. To characterize further its biochemical properties, we conducted studies in the bovine pituitary, an abundant source of native ICA512, as well as in fibroblasts transfected with various human ICA512 cDNA constructs. Based on these studies we have established that the signal peptide of ICA512 encompasses residues 1-34 and that the ectodomain of ICA512 undergoes multiple post-translation modifications, including N-glycosylation. Newly synthesized ICA512 appears first as a pro protein of 110 kDa that is then converted by post-translational modifications into a 130-kDa species. Cleavage of pro-ICA512 at a consensus for furin-like convertases generates a 60-66-kDa ICA512 transmembrane fragment (amino acids 449 979). Such processing ICA512 is not restricted to neuroendocrine cells, as it can also occur in transfected fibroblasts. Finally, the predicted N-terminal fragment of ICA512 resulting from this cleavage (amino acids 35-448) or parts thereof are present in the neurosecretosomes of posterior pituitary, raising the possibility that they may be secreted upon exocytosis of secretory granules. PMID- 10457161 TI - Cerebellar Golgi cells in the rat: receptive fields and timing of responses to facial stimulation. AB - Golgi cells are the only elements within the cerebellar cortex that inhibit granule cells. Despite their unique position there is little information on how Golgi cells respond to afferent input. We studied responses of Golgi cells to mechanical stimulation of the face, in Crus I-II of ketamine-xylazine anaesthetized rats. In 41 rats, 87 putative Golgi cells were identified, based on spike characteristics and on location of electrolytic lesions in the granular layer. They displayed a slow firing rhythm at rest (8.4 spikes/s). Most Golgi cells (84%) showed excitatory responses to tactile input. Their receptive fields (RFs) included, in 78%, the entire ipsilateral infraorbital nerve territory, and extended, in 14%, to other trigeminal nerve branches and, in 48%, to the contralateral face. Excitatory responses consisted of multiple, precisely timed (+/- 1 ms) spikes. Most peristimulus time histograms (PSTHs) (69%) showed an early (5-10 ms) and a late (13-26 ms) excitatory component, with each component consisting of a single PSTH peak. In some PSTHs the early component was a double peak (< 4 ms interval). In others, only one, early or late, PSTH peak was observed. The excitatory components were followed by a silent period (28-69 ms latency), the duration of which (13-200 ms) varied with response amplitude. In single cells, response profiles changed with stimulus location. In simultaneously recorded cells, evoked profiles differed for identical stimuli. Differences in RF size between early 'double' and 'single' peaks suggested that they resulted from direct mossy fibre and parallel fibre input, respectively. Late PSTH peaks were assumed to reflect corticopontine activation. PMID- 10457162 TI - Visual-field-specific heterogeneity within the tecto-rotundal projection of the pigeon. AB - The organization of the tecto-rotundal projection of the pigeon was investigated by means of anterograde and retrograde tracing techniques. Besides the known organization in tecto-rotundal connectivity, this study additionally demonstrates major variations in the ascending projections of different tectal subfields. We show that the ventral tectum opticum (TO) has significantly more projections onto the nucleus rotundus (Rt) than dorsal tectal areas. This difference coincides with differential innervation densities of afferent fibres within rotundal subregions. While ventral tectal efferents project onto the ventral and central Rt, dorsal tectal efferents mainly arborize within limited areas between the central Rt and its dorsal cap, the nucleus triangularis. Thus, the ventral TO, representing the lower and frontal field of view, exhibits a quantitatively and spatially enhanced projection onto the Rt, as compared with the dorsal TO. The data presented here demonstrate a visual field-dependent projection pattern of ascending tectal outputs onto different rotundal domains. The data are consistent with behavioural studies, demonstrating tectofugal lesions to suppress visual stimulus analysis mainly within the frontal field of view. PMID- 10457163 TI - Distribution, regulation and colocalization of the genes encoding the EP2- and EP4-PGE2 receptors in the rat brain and neuronal responses to systemic inflammation. AB - It is currently believed that prostaglandin (PG) of E2 type plays a crucial role in transferring the information received from circulating immune factors to brain parenchymal cells. Although PGE2 is synthesized quite essentially by cells of the blood-brain barrier, the organization and regulation of its receptor subtypes within neuronal elements remain unknown. In this study, intravenous (i.v.) injection of the endotoxin lipopolysaccharide (LPS) or recombinant rat interleukin-1beta (IL-1beta), and intramuscular (i.m.) injection of turpentine were used as different models of systemic immune stimuli. Rats were perfused at various times after the insults (30 min to 24 h), their brains cut and hybridized with full-length rat cRNA probes. Double-labelling procedures were accomplished to determine the cellular phenotype and activity. A very distinct distribution of both EP2 and EP4 receptors was found across the brain under basal conditions; the hybridization signal for the type 2 was detected in the bed nucleus of the stria terminalis (BNST), lateral septum, subfornical organ (SFO), ventromedial hypothalamic nucleus (VMH), central nucleus of the amygdala (CeA), locus coeruleus (LC) and the area postrema (AP), whereas the ventral septal/anterior preoptic area, the magnocellular paraventricular nucleus (PVN), supraoptic nucleus, parabrachial nucleus, LC, the nucleus of the solitary tract (NTS) and the ventrolateral medulla (VLM) exhibited moderate to strong levels for the EP4 mRNA under basal conditions. Upregulation of the genes encoding EP2 and EP4 receptors was detected in selective regions and neuronal populations during systemic inflammatory challenges. The most dramatic one being the robust transcriptional activation of the EP4 subtype within corticotropin-releasing factor (CRF) neurons of the parvocellular PVN following i.v. LPS and IL-1beta injection, and the localized i.m. aggression. These neurons of the endocrine hypothalamus as well as those of numerous autonomic-related nuclei were activated by the proinflammatory cytokine, as they were immunoreactive (ir) to Fos nuclear protein. The EP4 transcript was also present in activated catecholaminergic neurons of the LC, NTS and VLM, although only the A1 cell group exhibited an increase in EP4 transcription in response to circulating IL-1beta. Moreover, the systemic immunogenic insults caused a significant increase in the EP2 mRNA levels in the CeA, SFO, AP and the leptomeninges. These data provide a distinct pattern of EP2 and EP4 expression throughout the rat brain under both basal and immune challenged conditions, and underlie the possible role of the EP4 subtype in mediating the effects of PGE2 on different autonomic and neuroendocrine functions. The presence of Fos-ir nuclei in various populations of EP4 neurons of IL-1beta-treated animals clearly supports this concept and suggests that the selectivity of the neuronal response during systemic inflammation may depend on the expression of specific PGE2 receptors in key structures of the brain. PMID- 10457164 TI - The role of pattern vision in the development of cortico-cortical connections. AB - The development of cortico-cortical connections was studied in kittens deprived of vision by binocular eyelid suture during the formation of axonal arbors and synaptogenesis, i.e. between the second postnatal week and the end of the third postnatal month. Axons originating in area 17 and terminating either in ipsilateral or contralateral visual areas were visualized with biocytin. In ipsilateral areas 17 and 18, distinct clusters of branches begin to form, distally from the injection, during the second half of the first postnatal month, independently of pattern vision. More proximal clusters differentiate during the second postnatal month, and this seems to involve elimination of exuberant axonal branches. In kittens deprived of vision for 3 or more months, beginning before natural eye opening, the distal clusters regress and the proximal ones fail to differentiate. In extrastriate areas, distinct clusters of branches have segregated by the end of the second postnatal month, independently of visual experience; however, in kittens deprived of vision for 2 or more months, one of the clusters was selectively eliminated. In contralateral areas 17 and 18, we found stunted terminal arbors in kittens continuously deprived of vision. This was already noticeable at the end of the first postnatal month. Apparently, in the absence of pattern vision, most axons undergo only limited growth and do not form their characteristic terminal columns. Many of these axons are subsequently eliminated. In contrast, 8 days of vision beginning at natural eye opening and followed by visual deprivation caused a nearly normal development of intrahemispheric and interhemispheric connections. In conclusion, pattern vision appears to validate connections at early stages of their development; this validation is necessary for their further growth and differentiation that can then continue autonomously. PMID- 10457165 TI - An autoradiographic study of the distribution of binding sites for the novel alpha7-selective nicotinic radioligand [3H]-methyllycaconitine in the mouse brain. AB - [3H]-Methyllycaconitine ([3H]-MLA) is a new radioligand with selectivity for alpha7-type neuronal nicotinic acetylcholine receptors (nAChRs). In our previous study [Davies, A.R.L., Hardick, D.J., Blagbrough, I.S., Potter, B.V.L., Wolstenholme, A.J. & Wonnacott, S. (1999) Neuropharmacology, 38, 679-690], this radioligand labelled a single class of site in rat brain membranes; its pharmacology and distribution in crudely dissected brain regions closely paralleled that of the well-established alpha7-ligand [125I]-alpha-bungarotoxin. However, a small population of [3H]-MLA binding sites was apparently insensitive to alpha-bungarotoxin. Here we have extended the study to mouse brain, using autoradiography to examine the distribution of [3H]-MLA and [125I]-alpha bungarotoxin binding sites. [3H]-MLA labelled a single class of site in mouse brain membranes with a KD of 2.2 nM and a Bmax of 45.6 fmol/mg protein. Specific binding, defined by unlabelled MLA (Ki = 0.69 nM), was completely inhibited by ( )-nicotine (Ki = 1.62 microM), whereas alpha-bungarotoxin inhibited only 85% of specific binding (Ki = 3.5 nM). The distributions of [125I]-alpha-bungarotoxin and [3H]-MLA binding sites were compared by autoradiography, and binding was quantitated in 72 brain regions. Binding of both radioligands was highly correlated, with highest densities in the dorsal tegmental nucleus of the pons, colliculi and hippocampus. Serial sections labelled with [3H]-MLA in the absence or presence of unlabelled MLA or alpha-bungarotoxin provided no evidence for any alpha-bungarotoxin-resistant binding. The results are discussed in terms of binding sites that are inaccessible to alpha-bungarotoxin in membrane preparations. This study demonstrates the utility of [3H]-MLA for characterization of alpha7-type nicotinic receptors in mammalian brain, and suggests that it labels a population identical to that defined by [125I]-alpha bungarotoxin. PMID- 10457166 TI - Highly reproducible spatiotemporal patterns of mammalian embryonic movements at the developmental stage of the earliest spontaneous motility. AB - The principles underlying the variations in patterns of mammalian embryonic movements have not been established. In an attempt to clarify the mechanism that is responsible for the variations in motor patterns, we carried out a precise quantitative spatiotemporal analysis of movements in mouse embryos, using a transplacental perfusion method for the in vitro maintenance of live mammalian embryos. Episodes of spontaneous movements at the inception of motility, at embryonic day 12.5, occurred once every few minutes, lasted for several seconds and consisted of successive movements of body regions, the spatiotemporal patterns of which varied from episode to episode. By analysing and categorizing the patterns of these movements, we found that embryonic movements follow relatively few restricted patterns with respect to the order of the movements of body regions. A further analysis of episodes at high spatiotemporal resolution revealed that most of the episodes in a major category could be classified into two distinct subtypes. Each of these subtypes had its own highly reproducible spatiotemporal patterns of movement. Overall, these results show that early embryonic movements follow relatively few rather stereotyped patterns, and random local fluctuations have little effect on such movement patterns. The appearance of one pattern out of several rather stereotyped patterns may be the main cause of apparent variations in patterns of early embryonic movements. The stereotyped patterns may represent important orderly characteristics of spontaneous embryonic activities that may be involved in the development of orderly structures and functions in higher animals. PMID- 10457167 TI - Opioids modulate cell division in the germinal zone of the late embryonic neocortex. AB - Opioid effects on cell division in the embryonic cerebral cortex were examined using two experimental approaches: (i) the presence of opioid receptors in the embryonic day 16 mouse neocortex was tested using immunohistochemical techniques; (ii) the values of the indices of [3H]thymidine pulse labelled cells and mitotic indices were estimated in the ventricular zone of the embryonic day 16 mouse neocortex 2.5, 4.5 and 8.5 h after administration to pregnant females of selected opioid receptor agonists or the opioid antagonist naloxone. The immunohistochemical study demonstrated that distinct subpopulations of the ventricular zone cells express mu, delta or kappa opioid receptors. Acute exposure of mouse embryos to mu, delta and kappa opioid receptor agonists or naloxone differentially affects the indices of [3H] thymidine pulse labelled cells and mitotic indices indicating changes in the cell cycle composition. Treatment with the mu opioid receptor agonist D-Ala2-MePhe4, Gly-ol5-enkephalin (DAGO), or the partially selective kappa opioid receptor agonist bremazocine, increased the [3H]thymidine labelling and mitotic indices. In contrast, the delta receptor agonist (D-Ser8)-leucine enkephalin-Thr (DSLET) produced a decrease in the labelled cell indices and mitotic indices. Naloxone provided a biphasic effect: a decrease in the values of labelled cell indices 2.5 h after naloxone administration, followed by an increase in the values of the indices at 4.5 and 8.5 h. These results suggest that the endogenous embryonic/maternal opioid systems are involved in the regulation of cell division in the ventricular zone of the late embryonic cortex. PMID- 10457168 TI - A patchy horizontal organization of the somatosensory activation of the rat cerebellum demonstrated by functional MRI. AB - Blood oxygenation level dependent contrast (BOLD) functional MRI (fMRI) responses, in a 7-T magnet, were observed in the cerebellum of alpha-chloralose anaesthetized rats in response to innocuous electrical stimulation of a forepaw or hindpaw. The responses were imaged in both coronal and sagittal slices which allowed for a clear delineation and localization of the observed activations. We demonstrate the validity of our fMRI protocol by imaging the responses in somatosensory cortex to the same stimuli and by showing reproducibility of the cerebellar responses. Widespread bilateral activations were found with mainly a patchy and mediolateral band organization, more pronounced ipsilaterally. Possible parasagittal bands were observed only in contralateral lobule VI. There was no overlap between the cerebellar activations caused by forepaw and hindpaw stimuli. The overall horizontal organization of these responses was quite remarkable. For both stimulation paradigms most of the activation patches were positioned in either a rostral or caudal broad plane running anteroposteriorly through both anterior and posterior cerebellum. The rostral planes were completely separated, with the forepaw activation closer to the surface, while the caudal plane was common to both stimulation protocols. We relate our findings to the known projection patterns of spinocerebellar and cuneocerebellar mossy fibres, and to human fMRI studies. PMID- 10457169 TI - Cocaine self-administration increases the incentive motivational properties of the drug in rats. AB - A progressive increase in the frequency and intensity of drug use is one of the major behavioural phenomena characterizing the development of addiction. The nature of the drug-induced adaptations involved in this escalating drug intake remains unknown. Some theories propose that this escalation is due to a progressive decrease (tolerance) in the reinforcing or incentive effects of the drug. Alternative views posit that with chronic use the reinforcing or incentive effects of drugs increase, by a sensitization or a learning mechanism. In this report, we address the question of whether escalating cocaine intake is paralleled by an increase or a decrease in the reinforcing and incentive effects of the drug. Using the experimental model of intravenous drug self-administration with a within-session dose-response paradigm, we first studied the course of cocaine intake over 14 sessions in rats. After acquisition of cocaine self administration, cocaine intake progressively increased at each dose tested. Then rats, previously allowed to self-administer cocaine during either six or 29 sessions, were compared in three different tests of the incentive and reinforcing effects of cocaine: cocaine-induced reinstatement of self-administration, cocaine induced runway and cocaine-induced place conditioning. As compared with rats briefly exposed to cocaine self-administration (six sessions), rats with the longer experience (29 sessions) exhibited a higher intake of cocaine paralleled by a higher responsiveness in the cocaine-induced reinstatement and runway tests. Both groups of rats were similarly sensitive to the rewarding effects of the drug as evaluated by the threshold dose of cocaine inducing place conditioning. Our results demonstrate that escalating cocaine intake is paralleled by an increase in the motivational properties of the drug in the absence of apparent signs of tolerance to the reinforcing or incentive effects of cocaine. PMID- 10457170 TI - Presence of functional neuronal nicotinic acetylcholine receptors in brainstem motoneurons of the rat. AB - In mammals, nicotinic acetylcholine receptors (nAChRs) play a crucial role in motor control. Muscle-type nAChRs mediate synaptic excitation of skeletal muscle by motoneurons, and nAChRs are present on Renshaw cells, where they produce recurrent inhibition of spinal motoneurons. We asked whether nAChRs are also present in motoneurons. Whole-cell recordings were performed on various motor nuclei in brainstem slices of young rats. Neurons were visualized using infrared (IR) videomicroscopy. Acetylcholine (ACh) or the nicotinic agonist, epibatidine, were delivered by pressure microinjection. Facial (VII), hypoglossal (XII) and vagal (X) motoneurons responded to ACh by generating a fast inward current. In VII motoneurons, the ACh effect was mimicked by epibatidine, and nicotine induced a slow inward current and desensitized the ACh-evoked current. In VII and XII motoneurons, the ACh-evoked current was blocked by the nicotinic antagonist dihydro-beta-erythroidine (DHbetaE), but was unaffected by methyllycaconitine (MLA), an alpha7-specific antagonist. By contrast, the ACh-induced current in X motoneurons was sensitive to MLA. Current-voltage relationships indicated that the currents mediated by either alpha7-containing (X) or non-alpha7-containing (VII, XII) nAChRs displayed inward rectification. In accordance with the electrophysiological data, autoradiography revealed that VII, X and XII nuclei of young rats contained binding sites for [3H]epibatidine; binding sites for [125I]alpha-bungarotoxin, a selective ligand of alpha7-containing nAChRs, were present in X nucleus but were almost undetectable in VII and XII nuclei. Thus, brainstem motoneurons of young rats possess functional nAChRs. They could promote fast synaptic coupling between motoneurons, and thus play a role in somatic and visceral motor functions. PMID- 10457171 TI - Subthalamic nucleus lesions induce deficits as well as benefits in the hemiparkinsonian rat. AB - Lesions of the subthalamic nucleus can restore some imbalances in motor output of the basal ganglia induced by nigrostriatal dopamine depletion, and have been proposed as a potential therapy for Parkinson's disease. Although there is substantial supporting evidence from experimental studies in both rats and primates, there is less information on the effects of subthalamic lesions alone. In order to characterize potential side effects, the present study evaluates the behavioural effects of unilateral excitotoxic lesions of the subthalamic nucleus in rats that have previously received either unilateral saline or 6 hydroxydopamine injections into the nigrostriatal bundle on the same side. The 6 hydroxydopamine lesions induced ipsilateral orientation asymmetries in head position and body axis bias, rotational asymmetries following injections of direct or indirect dopamine agonists, neglect of contralateral stimuli, and a reduction in the numbers of pellets retrieved with the contralateral paw in a skilled reaching task. Subsequent excitotoxic lesions of the subthalamic nucleus reduced (but did not abolish) rotational asymmetries, had no effects on the measures of neglect and skilled paw-reaching, and produced contralateral orientation biases in head turning and body axis curling. Rats that received subthalamic lesions alone exhibited de novo impairments comprising contralateral biases in the orientation tests. These results support a neuromodulatory role of the subthalamic nucleus in regulating motor outputs of the basal ganglia, and caution that there may be distinct side effects of the lesion by itself. Whereas some impairments attributable to dopamine depletion may be alleviated by subthalamic manipulations, other symptoms are not, or may even be aggravated. PMID- 10457172 TI - Subpopulations of GABAergic and non-GABAergic rat dorsal horn neurons express Ca2+-permeable AMPA receptors. AB - Subpopulations of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors that are either permeable or impermeable to Ca2+ are expressed on dorsal horn neurons in culture. While both mediate synaptic transmission, the Ca2+ -permeable AMPA receptors provide a Ca2+ signal that may result in a transient change in synaptic strength [Gu, J.G., Albuquerque, C., Lee, C.J. & MacDermott, A.B. (1996) Nature, 381, 793]. To appreciate the relevance of these receptors to dorsal horn physiology, we have investigated whether they show selective expression in identified subpopulations of dorsal horn neurons. Expression of Ca2+-permeable AMPA receptors was assayed using the kainate-induced cobalt loading technique first developed by Pruss et al. [Pruss, R.M., Akeson, R.L., Racke, M.M. & Wilburn, J.L. (1991) Neuron, 7, 509]. Subpopulations of dorsal horn neurons were identified using immunocytochemistry for gamma aminobutyric acid (GABA), glycine, substance P receptor (NK1 receptor) and the Ca2+-binding proteins, calretinin and calbindin D28K. We demonstrate that, in dorsal horn neurons in culture, kainate-induced cobalt uptake is selectively mediated by Ca2+-permeable AMPA receptors, and that a majority of GABA and NK1 receptor-expressing neurons express Ca2+-permeable AMPA receptors. GABAergic dorsal horn neurons are important in local inhibition as well as in the regulation of transmitter release from primary afferent terminals. NK1 receptor expressing dorsal horn neurons include many of the projection neurons in the nociceptive spino-thalamic pathway. Thus, we have identified two populations of dorsal horn neurons representing important components of dorsal horn function that express Ca2+-permeable AMPA receptors. Furthermore, we show that several subpopulations of putative excitatory interneurons defined by calretinin and calbindin expression do not express Ca2+-permeable AMPA receptors. PMID- 10457173 TI - Functional and molecular diversity of PACAP/VIP receptors in cortical neurons and type I astrocytes. AB - In the present study we determined the mRNA-expression of pituitary adenylate cyclase activating polypeptide (PACAP)/vasoactive intestinal peptide (VIP) receptors in primary cultures of rat cortical neurons and type I astrocytes, and investigated the effects of PACAP38 on adenylyl cyclase, inositol phospholipid hydrolysis and intracellular calcium homeostasis. PACAP38 elicited a concentration-dependent (1 nM-100 nM) increase in inositol phosphate levels and [Ca2+]i in neurons but not in type I astrocytes. The PACAP-induced increase of intracellular calcium concentration, [Ca2+]i, was characterized by a spike, compatible with inositol trisphosphate (IP3) -induced calcium mobilization from intracellular stores, and a plateau phase, sustained by activation of capacitative calcium entry triggered by depletion of IP3-sensitive calcium stores. In the absence of extracellular calcium, only the spike phase was present while the plateau phase was clearly reduced. In addition, thapsigargin pretreatment abolished the PACAP38-induced [Ca2+]i rise. Treatment with 1 microM VIP did not affect [Ca2+]i in either neurons or type I astrocytes, clearly indicating the coupling of PAC1-HOP subtype to phospholipase-C in neurons. In addition, as previously reported, PACAP38 stimulated cAMP formation in both neurons and type I astrocytes. Using the reverse transcription polymerase chain reaction, we found mRNA-expression of PAC1 (PACAP - HOP variant) and VPAC2 in neurons, PAC1 (PACAP - R variant), VPAC1 and VPAC2 in astrocytes. These data indicate both a functional and molecular diversity of PACAP and VIP receptors in these cell types and support the view that the PAC1-HOP variant may be responsible for phospholipase-C activation and [Ca2+]i elevation in cortical neurons. PMID- 10457174 TI - Effects of GABAergic agonists and antagonists on oscillatory signal propagation in the guinea-pig accessory olfactory bulb slice revealed by optical recording. AB - To investigate the action of GABAergic agents on oscillatory signal propagation induced by electrical stimulation of the vomeronasal nerve layer, optical and electrophysiological recordings were carried out in slice preparations of the guinea-pig accessory olfactory bulb. In response to electrical stimuli, characteristic optical signals appeared in each layer: in the vomeronasal nerve layer, a transient presynaptic response; in the glomerular layer, pre- and postsynaptic responses; in the external plexiform, mitral cell and granule cell layers, a damped oscillatory response. Application of the GABAergic agonists, that is, GABA, muscimol (a GABAA receptor agonist) and baclofen (a GABAB receptor agonist), suggested that the GABAB action existed mainly in the glomeruli, whereas the GABAA action was present in both the glomeruli and the external plexiform layer. Bicuculline (a GABAA receptor antagonist) produced long-lasting but nonoscillating excitation in the external plexiform and mitral cell layers, indicating that the GABAA action contributes to the formation of oscillatory responses. When double-pulse stimulation was applied to the vomeronasal nerve layer, the test responses in the glomerular layer and external plexiform and mitral cell layers were depressed, but those in the vomeronasal nerve layer were not. Application of 2-hydroxysaclofen (a GABAB receptor antagonist) mostly blocked paired-pulse depression occurring in the glomerular layer and restored the reduced transmission to mitral cells, but had only a small effect on the depressed oscillatory response in the external plexiform and mitral cell layers. These observations suggest that GABAB action in the glomerular layer might, at least, regulate information flow from vomeronasal afferents to apical dendrites of mitral cells, like a gate inhibition. However, actions other than GABAB could also be involved in the depression of the oscillation in the external plexiform and mitral cell layers. PMID- 10457175 TI - The pH response of rat cutaneous nociceptors correlates with extracellular [Na+] and is increased under amiloride. AB - Excess hydrogen ions induce sustained nociceptor excitation as well as pain, and this has been suggested, with evidence from sensory ganglion cells, to result from gating a slowly inactivating sodium/calcium inward current. In the rat skin nerve preparation, isolated receptive fields of pH-sensitive C-fibre terminals were exposed to low-pH solutions of various sodium concentrations. The pH responses showed a good correlation with log [Na+]e, which supports the above model. Amiloride has previously been shown to block a pH-induced Na+ current involved in sensory transduction in hamster taste cells; however, it has been shown to act differently in cutaneous nociceptors. Amiloride induced a dose dependent increase in and prolongation of the nociceptive pH responses, with a prominent acceleration of the onset. The latter could be mimicked by replacing external sodium with sucrose, thus impeding sodium-proton antiport. Together, the findings indicate functional expression of amiloride-sensitive Na+/H+ antiporters, which enable the nociceptive nerve endings to extrude invading H+. Intracellular acidification may thus compete with Na+/H+ exchange, and pHi may be decisive in the transduction of nociception and pain from tissue acidosis. PMID- 10457176 TI - Astrocytes are required for the oscillatory activity in cultured hippocampal neurons. AB - Synchronous oscillations of intracellular calcium concentration ([Ca2+]i) and of membrane potential occurred in a limited population of glutamatergic hippocampal neurons grown in primary cultures. The oscillatory activity occurred in synaptically connected cells only when they were in the presence of astrocytes. Microcultures containing only one or a few neurons also displayed oscillatory activity, provided that glial cells participated in the network. The glutamate transporter inhibitors L-trans-pyrrolidine-2, 4-dicarboxylic acid (PDC) and dihydrokainate, which produce an accumulation of glutamate in the synaptic microenvironment, impaired the oscillatory activity. Moreover, in neurons not spontaneously oscillating, though in the presence of astrocytes, oscillations were induced by exogenous L-glutamate, but not by the stereoisomer D-glutamate, which is not taken up by glutamate transporters. These data demonstrate that astrocytes are essential for neuronal oscillatory activity and provide evidence that removal of glutamate from the synaptic environment is one of the major mechanisms by which glial cells allow the repetitive excitation of the postsynaptic cell. PMID- 10457178 TI - Visuospatial attentional functioning in mice: interactions between cholinergic manipulations and genotype. AB - Attentional functioning in mice was assessed in an analogue of the five-choice serial reaction time task in which the requirement was to detect brief visual stimuli presented across five spatial locations. Two hybrid strains of mice were assessed; F1 C57Bl/6xDBA/2 and C57Bl/6x129sv. Both strains acquired the task to high levels of performance with, in particular, no problems due to premature responding. At performance, systematic manipulation of the task parameters indicated a pattern of effects consistent with the task, taxing aspects of visuospatial attention. There were some differential effects of task manipulations at baseline across strain. However, the pattern of effects suggested these were likely to be the result of effects on factors other than attentional functioning per se, such as behavioural reactivity and inhibition. There was evidence in both strains of specific, centrally mediated effects of scopolamine on attentional functioning, with the C57Bl/6xDBA/2 hybrid showing greater sensitivity to the drug manipulation. Specific effects on discriminative accuracy were observed at doses of 0.02 and 0.2 mg/kg scopolamine. At the 2 mg/kg dose, large reductions in accuracy were associated with large effects on other measures, including omissions and response latencies, suggestive of nonspecific effects on task performance. These data indicate, for the first time, the utility of operant methods in assessing visuospatial attentional functioning in mice. They confirm the importance of cholinergic mechanisms in attentional processes across species, and suggest interactions between cholinergic mechanisms and genotype in the expression of attentional phenotypes. PMID- 10457177 TI - Acidic calponin immunoreactivity in postnatal rat brain and cultures: subcellular localization in growth cones, under the plasma membrane and along actin and glial filaments. AB - Acidic calponin, an F-actin-binding protein, is particularly enriched in brain, where calponin protein and mRNA are mainly expressed by neurons. The presence of calponin immunoreactivity in cultured astroglial cells has been reported, but the presence of acidic calponin in astrocytes in vivo appears equivocal. For the present study, we raised a specific polyclonal antibody against the 16-residue synthetic peptide covering the sequence E311-Q326 (EYPDEYPREYQYGDDQ) situated at the carboxy-terminal end of rat acidic calponin, and we investigated the cellular and subcellular localization of the protein in the developing central nervous system. Our results show that acidic calponin is particularly enriched in: (i) growth cones and submembranous fields of maturing cerebellar and cortical cells, where it codistributes with microfilaments and (ii) glial cells in vivo, including radial glia, glia limitans, Bergmann glia and mature astrocytes, and ex vivo, where acidic calponin strongly colocalizes with intermediate glial fibrillary acidic protein (GFAP) and vimentin filaments. Finally, up to four acidic calponin subtypes with different isoelectric point (pI) values were identified by two-dimensional gel electrophoresis of cerebellar and hippocampal extracts. The more acidic isoforms were developmentally regulated. As only one single mRNA for acidic calponin has been identified, these isoforms must reflect postsynthesis changes probably related to the particular functions of acidic calponin in maturing cells. Although brain acidic calponin's exact role remains uncertain, the present data suggest that it is involved in neuronal and glial plasticity. PMID- 10457180 TI - Partial denervation of the whiskerpad in adult mice: pattern and origin of reinnervation. AB - We studied sensory organ reinnervation after nerve transection in the mouse whisker-to-barrel pathway. In one set of adult mice, we determined at light microscopy level the number of fibres reaching the caudal whisker follicles 5, 15, 20, 60, 100 days and 1 year after transection of the sensory nerve of row C. Regenerated fibres were first detected 15 days post lesionem (p.l.) and myelin first observed at 20 days. Between 60 and 100 days, the number of fibres stayed at approximately 80% of the values obtained in control animals. At that time, myelinated fibres reached only 58% of their number in controls. At the electron microscopy level, these fibres differ from control ones by a smaller fibre diameter. The innervation of follicles of adjacent rows was not modified, indicating that follicular reinnervation is row specific. We checked this feature by injecting in another set of mice the denervated follicles and the adjacent ones with distinct retrograde tracers 45 days and 1 year after nerve transection. The percentage of double-labelled neurons in the Gasserian ganglion did not increase in experimental animals. This confirms the absence of colonization of intact follicles by regenerating fibres and indicates that reinnervation of the whisker follicles takes place by regeneration of the degenerated axons without collateral reinnervation. The companion paper describes the pattern of activation of the barrel cortex relative to the present findings. PMID- 10457179 TI - Effects of brain-derived neurotrophic factor on the development of NADPH diaphorase/nitric oxide synthase-positive amacrine cells in the rodent retina. AB - Amacrine neurons expressing nitric oxide synthase (NOS) contain brain-derived neurotrophic factor (BDNF) receptors and respond to exogenous BDNF [Klocker, N., Cellerino, A. & Bahr, M. (1998) J. Neurosci., 18, 1038-1046]. We analysed the effects of BDNF on the development of neurons which express NOS in the mouse and rat retina. Rat pups received a total of three intraocular injections of BDNF at intervals of 48 h, starting at postnatal day 16 (P16), and were killed at P22. The retinas were stained for NADPH-diaphorase, a histological marker of NOS. NOS expressing neurons were found in both the inner nuclear layer (INL) and the ganglion cell layer (GCL). Two classes of NOS-expressing neurons, type I and type II, had already been distinguished in the INL [Koistinaho, J. & Sagar, S.M. (1995) In Osborne, N.N. & Chader, G.J. (eds), Progress in Retinal and Eye Research, Vol. 15. Oxford University Press, pp. 69-87] and a third one in the GCL. Up-regulation of NADPH-diaphorase activity was observed after BDNF treatment. The number of type I neurons remained stable, whereas the number of type II neurons and NOS-positive neurons in the GCL increased significantly (P < 0.001). Type I and type II neurons were significantly larger in BDNF-treated retinas. Double-labelling experiments revealed that BDNF induces NADPH-diaphorase in dopaminergic neurons and amacrine cells displaced to the GCL, but not in retinal ganglion cells. In mice homozygous for a null mutation of the bdnf gene, the intensity of NADPH-diaphorase labelling in both somata and processes was reduced, but the number of labelled neurons was not dramatically reduced. These findings indicate that BDNF regulates the neurotransmitter phenotype of NOS expressing amacrine neurons under physiological conditions, but is not required for their survival. PMID- 10457181 TI - Partial denervation of the whiskerpad in adult mice: altered patterns of metabolic activity in barrel cortex. AB - One hundred days after unilateral C-row nerve transection in the adult mouse whiskerpad, the caudal follicles of row C are reinnervated with approximately 80 % of the original number of axons [Corthesy, M.-E., Bronchti, G. & Welker, E. (1999) Eur. J. Neurosci. , 11, 2835-2846]. To what extent is this reinnervation functional, and how does it interact with the enlargement of the functional representation of neighbouring rows subsequent to the denervation? Using the autoradiographic deoxyglucose method, we studied the whisker representation at the level of the barrel cortex 100 days post lesionem. We stimulated whiskers belonging to the denervated row C, the neighbouring rows B and D, or to all five rows A-E. The deoxyglucose uptake was measured in tangential sections through layer IV. The results indicate that, 100 days post lesionem, whiskers of row C reactivate their cortical barrels. However, (i) the magnitude of this cortical response was reduced; (ii) row C barrels were equivalently activated by the stimulation of the neighbouring rows; and (iii) when all whiskers were stimulated, we observed a significantly reduced deoxyglucose uptake over the representation of nonlesioned whiskers of rows D and E. Therefore, 100 days after the peripheral nerve lesion the reinnervation of the whiskerpad had not restored a normal pattern of activation at the level of the barrel cortex. We propose that this is due to a modified interaction between the representations of the various rows of follicles at the cortical level that does not return to normal. PMID- 10457182 TI - Expression of functional alpha7 nicotinic acetylcholine receptor during mammalian muscle development and denervation. AB - We have studied, on the transcriptional, protein and functional level, the expression of alpha7 nicotinic acetylcholine receptors (nAChR) in the course of rat muscle development, denervation and renervation. At foetal day 13, alpha7 nAChR expression was observed in somites and developing muscles of the back, but not yet in migrating myoblasts. Two days later, concomitant with myoblast aggregation, the alpha7 isoform began to be expressed in isolated myoblasts, with the highest level of expression in the frontal zone of the migrating wave. On foetal day 18, a time when the myoblasts in the upper hindleg have fused, alpha7 nAChR expression was most prominent in the outer layer of muscle tissue. The highest level of expression was observed in the first postnatal week, when practically all muscle cells stained positively for alpha7 protein. During the following weeks, alpha7 nAChR expression slowly decreased and practically disappeared in adult hindleg muscle. Following chronic denervation of adult Soleus muscle fibres, expression of alpha7 nAChR returned within 2-4 weeks. Electrophysiological measurements showed that the alpha7 nAChR of chronically denervated soleus muscle fibres were functional and, in particular, that they could be activated by choline. The presence of the alpha7 nAChR in developing and denervated muscle suggests a role for this nicotinic receptor in neuronal pathfinding and/or endplate stabilization. PMID- 10457183 TI - Different types of calcium channels and secretion from bovine chromaffin cells. AB - Bovine chromaffin cells possess several types of Ca2+ channels, and influx of Ca2+ is known to trigger secretion. However, discrepant information about the relative importance of the individual subtypes in secretion has been reported. We used whole-cell patch-clamp measurements in isolated cells in culture combined with fura-2 microfluorimetry and pharmacological manipulation to determine the dependence of secretion on different types of Ca2+ channels. We stimulated cells with relatively long depolarizing voltage-clamp pulses in a medium containing 60 mM CaCl2. We found that, within a certain range of pulse parameters, secretion as measured by membrane capacitance changes was mainly determined by the total cumulative charge of Ca2+ inflow and the basal [Ca2+] level preceding a stimulus. Blocking or reducing the contribution of specific types of Ca2+ channels using either 20 microM nifedipine plus 10 microM nimodipine or 1 microM omegaCTxGVIA (omega-conotoxin GVIA) or 2 microM omegaCTxMVIIC (omega-conotoxin MVIIC) reduced secretion in proportion to Ca2+ charge, irrespective of the toxin used. We conclude that for long-duration stimuli, which release a large fraction of the readily releasable pool of vesicles, it is not so important through which type of channels Ca2+ enters the cell. Release is determined by the total amount of Ca2+ entering and by the filling state of the readily releasable pool, which depends on basal [Ca2+] before the stimulus. This result does not preclude that other stimulation patterns may lead to responses in which subtype specificity of Ca2+ channels matters. PMID- 10457184 TI - Alternative splicing generates a novel isoform of the rat metabotropic GABA(B)R1 receptor. AB - Here we present a novel isoform of the metabotropic G-protein-coupled receptor for gamma-aminobutyric acid (GABA). The isoform, termed GABA(B)R1c (R1c), differs from the recently identified R1a and R1b receptors by an in-frame insertion of 31 amino acids between the second extracellular loop and the fifth transmembrane region. Analysis of the rat GABA(B)R1 gene demonstrates that the insertion is the result of an alternative splicing event within a 567-bp intron between exons 16 and 17. In situ hybridization in the rat brain shows a wide distribution of R1c transcripts and an overlap with the R1a and R1b transcripts. The highest mRNA levels are found in cerebellar Purkinje cells, cerebral cortex, thalamus and hippocampal CA1 and CA3 regions. Western blots and immunodetection of recombinant epitope-tagged receptors as well as [125I]CGP71872 photoaffinity labelling of cell membranes demonstrate that R1c is correctly expressed, although at a lower level than the previously identified isoforms. When coexpressed with the newly characterized GABA(B)R2, R1c functionally couples to G-protein-activated Kir3.1/3.2 channels in Xenopus oocytes and to PLC-activating chimeric G(alpha)qo subunits in HEK-293 cells with a similar EC50 for agonists. These data suggest that the R1c isoform represents a functional GABA(B)R in the rat brain. PMID- 10457185 TI - Expression of potassium channels during postnatal differentiation of rabbit Muller glial cells. AB - The postnatal maturation of Muller glial cells from immature radial glial cells is accompanied by specific changes in the activity of distinct types of K+ channels, as shown by whole-cell and cell-attached records on freshly isolated cells from retinae of young (postnatal days 1-30, P1-P30) and adult rabbits. (i) The density of inwardly rectifying currents, providing the main K+ conductance in adult Muller cells, was very low (0.8 pA/pF) from P1 to P6 but increased rapidly thereafter until a relatively stable level of 11.0 pA/pF was established at P17. (ii) Transient (A-type) K+ currents were expressed in all immature cells at a high density (9.6 pA/pF). After P12, both the percentage of cells with A-type currents and the peak amplitudes of the currents (2.8 pA/pF) declined. (iii) Delayed rectifying K+ currents developed slowly until after P30. (iv) The postnatal maturation of radial glial cells was accompanied by a strong decrease in the activity of large-conductance, Ca2+-activated K+ channels, the open probability of which (measured at the resting membrane potential) decreased from 0.69 at P2-4 to 0.06 at P13-14. The developmental decrease of the activity of Ca2+-activated K+ channels is assumed to be mainly caused by alteration of the resting membrane potential which developed from low values (-49 mV) at P1-6 to high adult values (-84 mV) after P13. The activity of each distinct type of K+ channel investigated is differently modulated by developmental regulation. This may reflect different functional requirements of immature and mature Muller cells. PMID- 10457186 TI - Subunit-selective modulation of GABAA receptors by the non-steroidal anti inflammatory agent, mefenamic acid. AB - Mefenamic acid (MFA) has anti-convulsant and pro-convulsant effects in vivo, and has been shown to potentiate and inhibit GABAA (gamma-aminobutyric acid) receptors in vitro. In this study, whole-cell currents were recorded from Xenopus oocytes and human embryonic kidney (HEK) cells expressing human recombinant GABAA receptors to resolve the molecular mechanisms by which MFA modulates GABAA receptor function. We demonstrate that MFA potentiated GABA-activated currents for alpha1beta2 gamma2S (EC50 = 3.2 +/- 0.5 microM), but not for alpha1beta1 gamma2S receptors. MFA also enhanced GABA-activated responses and directly activated alpha1beta2/beta3 GABAA receptors, but inhibited responses to GABA on alpha1beta1 constructs (IC50 = 40 +/- 7.2 microM). A comparison of beta1, beta2 and beta3 subunits suggested that the positive modulatory action of MFA involved asparagine (N) 290 in the second transmembrane domain (TM2) of the beta2 and beta3 subunits. Mutation of N290 to serine (S) markedly reduced modulation by MFA in alpha1beta2(N290S)gamma2S receptors, whereas alpha1beta1(S290N)gamma2S constructs revealed potentiated responses to GABA (EC50 = 7.8 +/- 1.7 microM) and direct activation by MFA. The potentiation by MFA displayed voltage sensitivity. The direct activation, potentiation and inhibitory aspects of MFA action were predominantly conferred by the beta subunits as the spontaneously active homomeric beta1 and beta3 receptors were susceptible to modulation by MFA. Molecular comparisons of MFA, loreclezole and etomidate, agents which exhibit similar selectivity for GABAA receptors, revealed their ability to adopt similar structural conformations. This study indicates that N290 in TM2 of beta2 and beta3 subunits is important for the regulation of GABAA receptor function by MFA. Our data provide a potential molecular mechanism for the complex central effects of MFA in vivo. PMID- 10457187 TI - Long-term effects of prenatal stress and postnatal handling on age-related glucocorticoid secretion and cognitive performance: a longitudinal study in the rat. AB - There is growing evidence that stress during prenatal and postnatal periods of life can modify adaptive capacities in adulthood. The hypothalamo-pituitary adrenal axis may mediate an animal's responses to perinatal stressful events and thus serve as a neurobiological substrate of the behavioural consequences of these early events. However, little is known about the long-term effects of prenatal stressors throughout the entire life of the animals. The focus of the present study was to examine the long-term influences of a prenatal and postnatal stress on glucocorticoid secretion and cognitive performance. Prenatal stress of rat dams during the last week of pregnancy and postnatal daily handling of rat pups during the first 3 weeks of life were used as stressors. The long-term effects of these manipulations were analysed using a longitudinal approach throughout the entire life of the animals, and were repeatedly tested in adulthood (4-7 months), middle age (13-16 months) and in later life (20-24 months). The study demonstrated that prenatal stress and postnatal handling induced opposite effects on both glucocorticoid secretion and cognitive performance. Prenatal stress accelerated the age-related hypothalamo-pituitary adrenal axis dysfunctions; indeed, circulating glucocorticoids levels of prenatally stressed middle-aged animals are similar to old control ones, and also induced cognitive impairments. In contrast, postnatal handling protected from the age-related neuroendocrine and behavioural alterations. These results show that the altered glucocorticoid secretion induced by early environmental manipulations is primary to the cognitive alterations observed only later in life and could be one cause of age-related memory deficits. PMID- 10457188 TI - A pre- and postsynaptic modulatory action of 5-HT and the 5-HT2A, 2C receptor agonist DOB on NMDA-evoked responses in the rat medial prefrontal cortex. AB - Intracellular recordings were made from pyramidal neurons in layers V and VI of the rat medial prefrontal cortex in slice preparations to investigate the effect of the serotonin 5-HT2A,2C receptor agonist (-)-1-2,5-dimethoxy-4-bromophenol-2 aminopropane (DOB) and 5-hydroxytryptamine (5-HT) on N-methyl-D-aspartate (NMDA) induced responses. Bath application of either DOB or 5-HT [in the presence of antagonists to 5-HT1A, 5-HT3 and gamma-aminobutytric acid (GABA) receptors] produced a concentration-dependent biphasic modulation of the NMDA responses. They facilitated and inhibited NMDA responses at low ( 40 gold particles/microm2) of GSH-immunoreactivity. Strong GSH immunoreactivity was also a typical feature of a subset of axon terminal- and axon fibre-like profiles in the aged rat that showed signs of axon dystrophy and degeneration. When comparing with normally appearing axon fibre profiles located in close vicinity, the population of aberrant axons had higher average levels of glutamate-immunoreactivity (+93%), and lower average levels of glycine immunoreactivity (-88%). No difference was seen regarding the levels of GABA. The results of this study lend support to the idea that aging in the spinal cord motor nuclei is associated with an increased oxidative stress and indicate that different transmitter systems are differentially affected by the degenerative process. PMID- 10457190 TI - Distribution of the voltage-dependent calcium channel alpha1G subunit mRNA and protein throughout the mature rat brain. AB - The molecular identity of a gene which encodes the pore-forming subunit (alpha1G) of a member of the family of low-voltage-activated, T-type, voltage-dependent calcium channels has been described recently. Although northern mRNA analyses have shown alpha1G to be expressed predominantly in the brain, the detailed cellular distribution of this protein in the central nervous system (CNS) has not yet been reported. The current study describes the preparation of a subunit specific alpha1G riboprobe and antiserum which have been used in parallel in situ mRNA hybridization and immunohistochemical studies to localize alpha1G in the mature rat brain. Both alpha1G mRNA and protein were widely distributed throughout the brain, but variations were observed in the relative level of expression in discrete nuclei. Immunoreactivity for alpha1G was typically localized in both the soma and dendrites of many neurons. Whilst alpha1G protein and mRNA expression were often observed in cells known to exhibit T-type current activity, some was also noted in regions, e.g. cerebellar granule cells, in which T-type activity has not been described. These observations may reflect differences between the subcellular distribution of channels that can be identified by immunohistochemical methods compared with electrophysiological techniques. PMID- 10457191 TI - Clonal expansion and cell dispersion in the developing mouse retina. AB - The present study has used two different approaches for labelling progenitor cells at the optic vesicle stage in order to examine patterns of clonal expansion and cellular dispersion within the developing retina. X-inactivation transgenic mice and chimeric mice expressing the lacZ reporter transgene were examined during development and in adulthood to study the radial and tangential dispersion of proliferating neuroepithelial cells and postmitotic retinal cells of known identities. Chimeric retinas were used to measure tangential dispersion distances, while transgenic retinas were used to assess the frequency of tangential dispersion for individual populations of retinal neurons. Tangential dispersion is shown to be a universal feature of particular retinal cell types, being contrasted with the strictly radial dispersion of other cells. Tangential dispersion is a relatively short-distance phenomenon, with distinct dispersion distances characteristic for cone, horizontal, amacrine and ganglion cells. Embryonic and postnatal retinas show that tangential dispersion occurs at different times for these distinct cell types, associated with their times of differentiation rather than their neurogenetic periods. These developmental results rule out the possibility that tangential dispersion is due to a passive displacement produced by the proliferation of later-born cells, or to the lateral dispersion of a dividing sibling; rather, they are consistent with the hypothesis that tangential dispersion plays a role in the establishment of the orderly spatial distribution of retinal mosaics. PMID- 10457192 TI - Effects of nerve growth factor on visual cortical plasticity require afferent electrical activity. AB - It is known that administration of nerve growth factor (NGF) prevents the ocular dominance shift induced by monocular deprivation in the rat. To determine whether electrical activity in the visual afferent pathway is required for NGF effects on ocular dominance, we infused NGF into the cortex of animals subjected to complete monocular blockade of retinal discharges. Rats at the peak of the critical period received intravitreal tetrodotoxin (TTX) injections to silence activity in one eye for a period of 6-7 days; NGF was concurrently delivered into the visual cortex by means of osmotic minipumps. At the end of the treatment period, the ocular dominance distribution of cortical neurons was assessed by single-cell recordings. The results demonstrate that while infusion of NGF is effective in preventing the ocular dominance shift in lid-sutured rats, virtually no rescue can be observed in TTX-injected animals. Identical results were obtained when a specific agonist of the NGF receptor TrkA, the bivalent anti-rat TrkA IgG (RTA), was infused into the cortex in place of NGF. We conclude that NGF signalling via the TrkA receptor must be coupled to afferent electrical activity to produce its effects on the eye preference of cortical neurons. This suggests a generalized mechanism in which high-affinity neurotrophin receptor activation and afferent discharge interact to modulate neuronal plasticity in the developing visual cortex. PMID- 10457193 TI - Re-examination of the human taste region: a positron emission tomography study. AB - There is considerable uncertainty regarding the cortical areas in the human brain that are involved in gustatory processing. Evidence from nonhuman primates indicates that parts of the peri-central opercular region (secondary somatosensory cortex) and insular cortex may be important for gustatory processing. The aim of the study was to examine changes in cerebral blood flow during gustatory stimulation (with sucrose or water) in the insulo-opercular region of the human brain with positron emission tomography using only movement of the tongue and mouth as control conditions. This is important because subtractions of responses to one gustatory stimulus from those to another may mask gustatory activity that is common to both stimuli, even when the control stimulus is an apparently tasteless one (e.g. water). Bilateral increases in activity were observed in the insulo-opercular region and, consistent with animal work, they indicate that there are a number of separate foci within this general area where primary gustatory inputs may be processed. PMID- 10457194 TI - Calcium channel heterogeneity among cone photoreceptors in the tree shrew retina. AB - Retinal photoreceptors are depolarized in darkness and release neurotransmitter tonically. They respond to light by hyperpolarization and a concomitant reduction in transmitter release. The calcium-dependent release of transmitter is coupled to graded changes in membrane potential by L-type calcium channels in the photoreceptor terminals. This paper reports the immuno-localization of an L-type channel alpha1D subunit to most, but not all, synaptic terminals of cones in the tree shrew retina. Double labelling for the alpha1D subunit and the plasma membrane Ca2+-ATPase, which has been shown to be present in all tree shrew cones, revealed a subpopulation of cone terminals that did not react with the alpha1D antibody. The nonimmunoreactive synaptic terminals represented approximately 5.8% of the total and formed a highly regular array across the retina reminiscent of the blue cones. Double-staining for the alpha1D subunit and blue cone opsin confirmed that these are the blue cones. The observed differences in calcium channel immunoreactivity between long and short wavelength cones points to previously unsuspected heterogeneity in the molecular machinery governing transmitter release from spectrally different cone types. PMID- 10457195 TI - Interactions of HIV-1 with antigen-presenting cells. AB - There is currently much interest in the numerical and functional loss of antigen presenting cells (APC) in HIV-1 disease and the contribution that this may make to HIV-1 pathology. The HIV-1 virus can interfere with the normal function of APC in a number of ways involving inappropriate signalling. These include changes in cytokine balance, cell-surface molecule expression and intracellular signalling pathways. This review examines how HIV-1 is able to disregulate APC function and discusses possible outcomes for the function of the immune system. PMID- 10457197 TI - Cell death induced by the Fas/Fas ligand pathway and its role in pathology. AB - Engagement of the cell death surface receptor Fas by Fas ligand (FasL) results in apoptotic cell death, mediated by caspase activation. Cell death mediated via Fas/FasL interaction is important for homeostasis of cells in the immune system and for maintaining immune-privileged sites in the body. Killing via the Fas/FasL pathway also constitutes an important pathway of killing for cytotoxic T cells. Fas ligand is induced in activated T cells, resulting in activation-induced cell death by the Fas/FasL pathway. Recently it has been shown that the Fas receptor can also be up-regulated following a lesion to the cell, particularly that induced by DNA-damaging agents. This can then result in killing of the cell by a Fas/FasL-dependent pathway. Up-regulation of Fas receptor following DNA damage appears to be p53 dependent. PMID- 10457196 TI - The role of B7 costimulation in T-cell immunity. AB - CD4+ T cells are considered to be the major controlling element of the adaptive immune response. They recognize foreign peptides by interaction of the T cell receptor (TCR) with peptide complexed to major histocompatibility complex (MHC) class II molecules on the surface of antigen presenting cells (APC). Once activated, CD4+ T cells orchestrate the various phases of the immune response. They are responsible for the production of numerous cytokines, which activate specific immune effector cell populations including B cells, eosinophils, mast cells and macrophages. Not surprisingly, the activation of CD4+ T cells needs to be tightly regulated and is subject to finely tuned control mechanisms. The requirement for a second or 'costimulatory' signal, in addition to the antigenic signal, provides a key element for the exquisite control of T cell activation. One of the major signalling pathways responsible for delivery of this costimulatory signal is induced by interaction of CD28 on T cells with B7 molecules found only on APC. The present review outlines our current understanding of the physiological role of B7 costimulatory signals in regulating CD4+ T cell responses. PMID- 10457198 TI - Inhibition of naive class I-restricted T cells by altered peptide ligands. AB - Amino acid variants of an antigenic peptide or altered peptide ligands have previously been investigated with CD4+ and CD8+ T cells. However, for CD8+ T cells, only clones (which are continually restimulated in vitro) have been assessed. Using TCR transgenic mice specific for a class I Kb-restricted OVA peptide (OVAp; OT-I mice) as a source of naive CD8+ T cells, single amino acid variants of the OVAp were analysed in vitro for their ability to antagonize the proliferative and cytotoxic function of naive OT-I cells. Peptides with substitutions at TCR contact residues were found to be the most potent antagonists of OT-I cell function. Those peptides that inhibited activation of cells to proliferate also inhibited activation of cells to become killers. Inhibition was inversely correlated with interferon (IFN)-gamma production. It was found that levels of antagonist peptide required for inhibition were higher than that described for T cell clones, presumably due to affinity differences. PMID- 10457199 TI - Use of recombinant viruses to deliver cytokines influencing the course of experimental bacterial infection. AB - The feasibility of using viral constructs expressing cytokine genes to influence the course of bacterial infection was tested in mice. The mice were first infected with vaccinia or fowlpox viruses expressing the cytokine of interest, then challenged with the facultative intracellular bacterial pathogen Listeria monocytogenes. The course of infection was assessed by subsequent bacterial counts. Expression of IFN-gamma or TNF was protective. Vaccinia virus was more efficient at delivering IFN-gamma-mediated protection than was fowlpox virus, which is unable to proliferate in mammalian cells. The effect of vaccinia-IFN gamma was more apparent in the liver, where vaccinia proliferates to high titres (> 109), than in the spleen, where only 103 vaccinia were isolated. Vaccinia virus expressing IL-4 exacerbated infection. Interleukin-4 exacerbation was T cell independent and was reflected in the failure of macrophage activation, possibly due to suppression of NK cells, which are a source of IFN-gamma early in infection. The clear indication of protection by some cytokines in this prophylactic model appears to justify further study of the therapeutic effects of cytokine-expressing viruses in chronic bacterial infections, especially where a cytokine defect is suspected. PMID- 10457201 TI - Mouse M290 is the functional homologue of the human mucosal lymphocyte integrin HML-1: antagonism between the integrin ligands E-cadherin and RGD tripeptide. AB - Human mucosal lymphocyte antigen-1 (HML-1, alphaEbeta7) and E-cadherin, two members of unrelated cell adhesion superfamilies, have evolved to play cooperative roles in gut mucosal immunity. Human E-cadherin is self-ligand mediating intercellular adhesion of epithelial cells, as well as adhesion of intra-epithelial lymphocytes to intestinal enterocytes via an interaction with HML-1. Herein we report that both dimeric and monomeric forms of recombinant mouse E-cadherin-human immunoglobulin Fc chimera self-associate and support attachment of E-cadherin+ mouse colon epithelial cells. Both forms also support the adhesion of mouse MTC-1 T cells via M290, thereby establishing M290 as the functional mouse homologue of HML-1 and revealing that E-cadherin homophilic and heterophilic binding sites are distinct. Adhesion of MTC-1 cells to E-cadherin-Fc was inhibited by arginine-glycine-aspartate (RGD) peptides and vice versa cells bound to immobilized RGD polymer in an M290-dependent fashion, where adhesion was inhibitable with soluble E-cadherin-Fc. Hence, E-cadherin and RGD integrin ligands antagonize cell binding by one another, either by inducing integrin cross talk or by binding to shared or overlapping sites within M290. Binding of E cadherin-Fc by HML-1 costimulated the CD3-induced proliferation of purified CD4+ T cells, suggesting that E-cadherin expressed on dendritic cells may play a T cell costimulatory role in addition to facilitating dendritic cell-keratinocyte adhesion. PMID- 10457200 TI - Characterization of a bioassay for detection of recombinant and native ovine interleukin-5. AB - In order to analyse Th2-type immune responses in sheep by the assay of interleukin (IL)-5 in biological fluids, the ovine IL-5 gene was cloned and expressed in Spodoptera frugiperda Sf9 insect cells using the baculovirus expression vector system. The recombinant product was purified as BAC-OV-IL-5 from the supernatant fluid. The ovine IL-5 was biologically active in a bioassay using IL-5-dependent Baf cells, which have been used previously to specifically detect human IL-5. The specificity of Baf cells for ovine IL-5 was examined by two methods. First, Baf cells only proliferated in response to BAC-OV-IL-5 and did not respond to addition of recombinant ovine cytokines granulocyte-macrophage colony stimulating factor (GM-CSF), IL-1beta, IL-2, IL-3, IL-6, IL-8, stem cell factor (SCF) or IFN-gamma at doses from 0.01 to 1 microg/well. Second, the rat monoclonal antibody to murine IL-5, TRFK-5, neutralized murine, but not ovine, IL 5. However, rabbit antisera to BAC-OV-IL-5 neutralized murine and ovine recombinant IL-5 and abolished responses of Baf cells to IL-5 activity in supernatant fluids from mesenteric lymph node cells (MLNC) of parasitized sheep. The bioassay had a sensitivity to detect 8 ng in a 200 microL assay (40 ng/mL). Thus, the specificity of Baf cells to detect human IL-5 also extends to ovine IL 5 and therefore provides a method for monitoring the production of Th2 immune reactivity in sheep. PMID- 10457202 TI - Modulation of immune function by a modified bovine whey protein concentrate. AB - The commercial preparation of dairy foodstuffs generates large volumes of by products, many of which have as yet undocumented effects on mammalian immune function. In the present report, a modified whey protein concentrate (mWPC), derived as a by-product from the commercial manufacture of cheese, was tested for its ability to modulate murine immune function in vitro. The mWPC suppressed T and B lymphocyte proliferative responses to mitogens in a dose-dependent fashion. The mWPC also suppressed alloantigen-induced lymphocyte proliferation during a mixed leucocyte reaction, but showed no suppressive effect against IL-2-sustained proliferation of mitogen-activated T cell blasts. Other indices of lymphocyte activation, such as cytokine secretion and the formation of activated (CD25+) T cell blasts, were suppressed by the mWPC, suggesting that the mode of suppression may be to inhibit the lymphocyte activation process. Enzymatic digestion by pepsin and pancreatin, under physiologically realistic conditions in vitro, ablated the immunomodulatory function of the mWPC. These results are discussed in relation to the potential development of complex-mixture dairy products into health-modulating products. PMID- 10457203 TI - Salivary IgA subclasses and infection risk in elite swimmers. AB - The concentrations of total IgA, IgA1 and IgA2 were measured in saliva collected from 25 elite swimmers in the early and late phases of a 7 month training season and compared with the number of respiratory infections during the season. The IgA1 concentrations in the early phase of the training season were significantly associated (P = 0.01) with the number of respiratory infection episodes during the training season. The lower the concentration of IgA1, the greater the number of infection episodes. Swimmers with four or more infections during the training season had significantly lower salivary IgA1 concentrations than those with less than four infection episodes (P = 0.01). The proportion of IgA1 in the saliva of the elite swimmers (80%) was higher than for normal non-exercising adults (60%). A small proportion of athletes had salivary IgA2 concentrations below the detection limit of the assay and the mean concentration of IgA2 was significantly lower than the concentrations for a normal adult population (P = 0.01). This study suggests that measurement of IgA subclasses, in particular IgA1, at the commencement of a training season may predict infection risk in elite swimmers. PMID- 10457205 TI - Mucosa-associated lymphoid tissues as sites for uptake, carriage and excretion of tubercle bacilli and other pathogenic mycobacteria. AB - Pathogenic mycobacteria, including those that cause tuberculosis and paratuberculosis, cross mucosal barriers by endocytosis within mucosal lymphoepithelial sites. These entry sites commonly include oropharyngeal and nasopharyngeal tonsils and Peyer's patches. Bacilli discharged at the basolateral surfaces of engulfing epithelial M cells are taken up by professional antigen presenting cells associated with T lymphocytes of the parafollicular area. Dendritic cells and macrophages in these sites allow mycobacterial replication, due to the permissive immunological environment in lymphoepithelial tissues. Abrogation of local delayed-type hypersensitivity reactions generally ensures continuing integrity and function of these tissues. Phagocytes containing intracellular mycobacteria disseminate infection to other parts of the body and also probably migrate back onto the mucosal surface to shed bacilli. PMID- 10457204 TI - Expression and activation of RAS and mitogen-activated protein kinases in macrophages treated in vitro with cisplatin: regulation by kinases, phosphatases and Ca2+/calmodulin. AB - Cisplatin (cis-dichlorodiammineplatinum II), a potent antitumour compound, stimulates immune responses by activating monocytes/macrophages and other cells of the immune system. However, the exact mechanism by which cisplatin activates these cells is poorly characterized and attempts are being made to understand this mechanism. Previous studies from this laboratory have shown that Lyn, a protein tyrosine kinase of the src family, and nuclear factor (NF)-kappaB are involved in cisplatin-induced macrophage activation. Recent studies suggest that the RAS and mitogen-activated protein (MAP) kinases function as a connecting link between activated lyn and NF-kB, which raises the possibility of their involvement in cisplatin-induced macrophage activation. Therefore, this study was undertaken to investigate the effect of cisplatin treatment on the expression/activation of RAS (a low molecular weight GTP-binding protein) and MAP kinases in murine peritoneal macrophages. The underlying mechanism of expression/activation of RAS and MAP kinases in cisplatin-treated macrophages was also investigated. Immunoblotting and immune-complex kinase assays revealed that cisplatin treatment of macrophages leads to increased expression/activation of RAS and MAP kinases, with optimal expression/activation at 15 min of treatment. Using a battery of specific inhibitor/modulators of different signalling molecules, this study shows that expression and activation of MAP kinases are two unrelated processes. It was also observed that kinase (protein tyrosine and protein kinase C) inhibitor and Ca2+/calmodulin antagonist inhibit expression/activation of RAS/MAP kinases in macrophages, whereas phosphatases (protein tyrosine and serine/threonine) inhibitor up-regulate these kinases. PMID- 10457206 TI - Concanavalin A stimulation enhanced secondary VlambdaJlambda rearrangement in some human plasma B cells without up-regulation of recombination-activating gene expression and Vlambda germline transcription. AB - Light chain shifting is a phenomenon that occurs in certain human antibody producing plasma B cells which, when stimulated with concanavalin A (Con A), shift production of the original light chain to new light chains. Here we investigated the effect of Con A stimulation on these light chain shift-inducible cells. Analysis of transcripts and VJ-coding joints for new light chains revealed that a leaky amount of secondary VlambdaJlambda rearrangement occurs spontaneously, without Con A stimulation, and that Con A stimulation markedly increases VJ-coding joints and transcripts for new light chains. It was also shown that new light chain producers, which have carried out secondary rearrangement, do not further rearrange their light chain genes, even when stimulated with Con A. Recombination-activating gene (RAG) products and Vlambda germline transcription were constitutively expressed in these cell lines and their expression levels were not affected by Con A stimulation. These results suggest that Con A stimulation enhanced secondary VlambdaJlambda rearrangement, but this was not a result of the up-regulation of RAG expression and Vlambda germline transcription, which are believed to be sufficient for the process of VlambdaJlambda rearrangement. PMID- 10457207 TI - Characteristics of IgVH genes used by human intestinal plasma cells from childhood. AB - Plasma cells secreting immunoglobulin M (IgM) and IgA in human intestinal mucosa are the largest antibody-producing population in the human body. Despite this there have been relatively few studies of the characteristics and maturation of the genes which encode the mucosal immunoglobulins. We have previously demonstrated that intestinal plasma cells use highly mutated IgVH genes, likely to reflect germinal centre origin. Here we show that IgVH genes used by intestinal lamina propria plasma cells secreting IgM and IgA are highly mutated from childhood, with no change in the frequency of mutation through to adulthood, though IgVH genes used by IgM are significantly less mutated than those used by IgA. There was no difference between the IgA subclasses in either the frequency or distribution of mutations. The frequency of mutation in IgVH4-34 genes used by IgG was also studied in the adult biopsies, and was found to be of the same order as that observed in IgA and was significantly higher than that observed in IgM. We have identified IgM and IgA sequences which share identical CDR3 and distribution of mutations. Isotype switching may therefore occur after extensive mutation of IgM sequences, and IgM- and IgA-secreting plasma cells with the same specificity may occur within the same microenvironment. IgM should therefore be considered to be a component of secondary immune responses in the gut. PMID- 10457208 TI - Visualizing the T-cell response elicited by oral administration of soluble protein antigen. AB - Oral administration of soluble protein antigen induces tolerance, while particulate antigens encountered in the intestine provoke active immunity. Although the events that lead to these distinct outcomes are not yet fully characterized, they may reflect differences at the antigen-presenting cell (APC) level. The role of dendritic cells (DC) in regulating responses at mucosal sites has remained largely undefined because of the low frequency of DC in mucosal associated tissues. In this study we have used the growth factor Flt3-ligand (Flt3L) to expand DC populations in vivo, in combination with an adoptive transfer system, in order to track antigen-specific T cells during oral tolerance induction. We observed rapid T-cell activation, localized particularly in the mucosal tissues, within hours after feeding the soluble protein antigen, ovalbumin (OVA). The response was enhanced in Flt3L-treated mice, indicating an important role for DC during the inductive phase of tolerance. PMID- 10457209 TI - Characterization of the memory/activated T cells that mediate the long-lived host response against tuberculosis after bacillus Calmette-Guerin or DNA vaccination. AB - The memory/activated T cells, which mediate the long-lived host response against tuberculosis, in mice immunized with either bacillus Calmette-Guerin (BCG) or mycobacterium heat-shock protein 65 (hsp 65) antigen expressed from plasmid DNA (DNA-hsp 65), were characterized. Protection against Mycobacterium tuberculosis challenge by DNA-hsp 65 vaccination was associated with the presence of lymph node T-cell populations in which CD8+/CD44hi interferon-gamma (IFN-gamma) producing/cytotoxic cells were prominent even after 8 or 15 months of plasmid DNA mediated immunizations, whereas after BCG vaccination the majority were CD4+/CD44lo IFN-gamma-producing T cells. When the cells were separated into CD4+CD8- and CD8+CD4- and then into CD44hi and CD44lo types, CD44lo cells were essentially unable to transfer protection in adoptive transfer experiments, the most protective CD44hi cells were CD8+CD4- and those from DNA-vaccinated mice were much more protective than those from BCG-immunized mice. The frequency of protective T cells and the level of protection were increased up to 8 months and decreased after 15 months following DNA or BCG immunizations. PMID- 10457210 TI - Local effects of recombinant rat interleukin-6 on the peripheral nervous system. AB - Interleukin-6 (IL-6) is a multifunctional cytokine with a broad range of activities and can affect a variety of target cells or systems in multiple ways. However, there is currently no consensus on how IL-6 directly affects the peripheral nervous tissue. We performed histopathological and immunohistochemical analyses to investigate the direct effects of recombinant rat IL-6 (rrIL-6) following its intraneural injection into the sciatic nerve of adult Lewis rats. One day after injection, a large number of macrophages, major histocompatibility complex (MHC) class II positive cells, and CD4+ and CD8+ T cells appeared within the perineurium and endoneurium. From day 4 to day 7 after injection, we observed a gradual increase of inflammation and demyelination. On day 7, demyelination affected more than 80% of nerve fibres. In contrast, in the sterile phosphate buffered saline (PBS)-injected control group, lower inflammation and fewer demyelinating nerve fibres were observed on days 4 and 7. Thus, intraneural injection of rrIL-6 into the sciatic nerve induces high inflammation and severe demyelination. This study improves our understanding of the effector mechanisms underlying inflammation and demyelination and identifies IL-6 as an essential mediator of inflammation and demyelination in the peripheral nervous system after local administration. PMID- 10457211 TI - Interleukin-12 can directly induce T-helper 1 responses in interferon-gamma (IFN gamma) receptor-deficient mice, but requires IFN-gamma signalling to downregulate T-helper 2 responses. AB - An in vivo model of pulmonary granuloma formation around embolized schistosome eggs was investigated as an environment in which to analyse a role for interleukin-12 (IL-12) in the differentiation of T-helper 1 (Th1) and Th2 subsets. Specifically, mice deficient for the interferon-gamma receptor (IFN gammaR-/-) were used to determine the role for IL-12 in the absence of IFN-gamma mediated signalling. We show that recombinant IL-12 administered to IFN-gammaR-/- mice caused the up-regulation of mRNA for IFN-gamma in lung tissue, and the secretion of abundant IFN-gamma by in vitro-cultured lymph node cells in response to egg antigens. This indicates that IL-12 can act independently of IFN-gamma to induce the development of Th1 cells. Administration of rIL-12 to wild-type mice markedly reduced the secretion of Th2-associated cytokines, IL-4 and IL-5. However, these cytokines were not dramatically reduced in IFN-gammaR-/- mice treated with IL-12. We conclude that inhibition of these cytokines by IL-12 is primarily dependent upon effective IFN-gamma signalling, although abrogation of T cell derived IL-10 appeared to be dependent upon IL-12. We also show that increases in mRNA for the beta2 subunit of the IL-12 receptor and the p40 subunit of IL-12 after rIL-12 treatment were lower in IFN-gammaR-/- mice, compared to wild-type mice, indicating that their expression was primarily dependent upon IFN gamma with only a minor role for IL-12. PMID- 10457212 TI - Milk cytokines and subclinical breast inflammation in Tanzanian women: effects of dietary red palm oil or sunflower oil supplementation. AB - Previously, we have found that subclinical breast inflammation, as indicated by raised breastmilk concentrations of sodium and the inflammatory cytokine, interleukin-8 (IL-8), was highly prevalent in Bangladesh and associated with poor infant growth. In order to investigate further the prevalence of subclinical breast inflammation and to assess the impact of dietary intervention, we studied rural Tanzanian women taking part in a study of dietary sunflower or red palm oil supplementation during late pregnancy and lactation. We measured breastmilk concentrations of IL-8, the anti-inflammatory cytokine, transforming growth factor-beta2 (TGF-beta) and the ratio of sodium to potassium. We also estimated systemic inflammation by plasma concentrations of the acute phase proteins, alpha1-acid glycoprotein and C-reactive protein. There were highly significant intercorrelations among milk Na/K ratio and concentrations of IL-8 and TGF-beta, the last only after treatment with bile salts which also improved TGF-beta recovery in the enzyme-linked immunosorbent assay (ELISA). Plasma acute phase protein concentrations tended to correlate with milk Na/K ratio and IL-8, suggesting that subclinical breast inflammation was related to systemic inflammation. Dietary supplementation with vitamin E-rich sunflower oil but not provitamin A-containing red palm oil decreased milk Na/K, IL-8 and TGF-beta at 3 months postpartum; however, the effect was significant only for Na/K ratio. The results suggest that milk Na/K ratio, IL-8, and TGF-beta all measure the same phenomenon of subclinical breast inflammation but that Na/K ratio, having the lowest assay variability, is the most useful. Subclinical breast inflammation may result in part from systemic inflammation and may be improved by increased dietary intake of vitamin E-rich sunflower oil. PMID- 10457213 TI - Different effect of granulocyte colony-stimulating factor or bacterial infection on bone-marrow cells of cyclophosphamide-treated or irradiated mice. AB - In the present study, the effect of treatment with granulocyte colony-stimulating factor (G-CSF) on cellular composition of the bone marrow and the number of circulating leucocytes of granulocytopenic mice, whether or not infected with Staphylococcus aureus, was assessed. With two monoclonal antibodies, six morphologically distinct cell populations in the bone marrow could be characterised and quantitated by two-dimensional flow cytometry. Granulocytopenia was induced by cyclophosphamide or sublethal irradiation. Cyclophosphamide predominantly affected the later stages of dividing cells in the bone marrow resulting in a decrease in number of granulocytic cells, monocytic cells, lymphoid cells and myeloid blasts. G-CSF administration to cyclophosphamide treated mice increased the number of early blasts, myeloid blasts and granulocytic cells in the bone marrow, which indicates that this growth factor stimulates the proliferation of these cells in the bone marrow. During infection in cyclophosphamide-treated mice the number of myeloid blasts increased. However, when an infection was induced in cyclophosphamide and G-CSF-treated mice, the proliferation of bone-marrow cells was not changed compared to that in noninfected similarly treated mice. Sublethal irradiation affected all bone marrow cell populations, including the early blasts. G-CSF-treatment of irradiated mice increased only the number of myeloid blasts slightly, whereas an infection in irradiated mice, whether or not treated with G-CSF, did not affect the number of bone-marrow cells. Together, these studies demonstrated that irradiation affects the early blasts and myeloid blasts in the bone marrow more severely than treatment with cyclophosphamide. Irradiation probably depletes the bone marrow from G-CSF-responsive cells, while cyclophosphamide spared G-CSF responsive cells, thus enabling the enhanced G-CSF-mediated recovery after cyclophosphamide treatment. Only in these mice, bone marrow recovery is followed by a strong mobilisation of mature granulocytes and their band forms from the bone marrow into the circulation during a bacterial infection. PMID- 10457214 TI - Macrophages expressing heat-shock protein 65 play an essential role in protection of mice infected with Plasmodium yoelii. AB - C57BL/6 (B6) mice are resistant to infection with the non-lethal (NL) strain of Plasmodium yoelii 17X, while being susceptible to that with the lethal (L) strain. The 65 000 MW heat-shock protein (hsp 65) was strongly expressed in splenic adherent cells of B6 mice 10 days after infection with the NL strain of P. yoelii but only slightly in those from mice infected with the L strain. Mice which had survived infection with the NL strain were resistant to challenge with the L strain and hsp 65 was strongly expressed in splenic adherent cells of these mice. Severe combined immunodeficient mice and nude mice were susceptible to malaria infection even with the NL strain and did not express hsp 65 after infection, suggesting that T cells are required for the expression of hsp 65 as well as for protective immunity. B6 mice treated intraperitoneally with carrageenan, which impairs the macrophage function, became susceptible to NL strain infection, indicating that macrophages play an important role as the final effectors in protective immunity. These results demonstrate that the hsp 65 expressed by macrophages correlates closely with protection against P. yoelii infection. PMID- 10457215 TI - Therapy of established tumour with a hybrid cellular vaccine generated by using granulocyte-macrophage colony-stimulating factor genetically modified dendritic cells. AB - Dendritic cells (DCs) are the most powerful of all antigen-presenting cells and play a critical role in the induction of primary immune responses. DC-based vaccination represents a potentially powerful strategy for cancer immunotherapy. In this study, a new approach for a DC-based melanoma vaccine was described. Splenic DCs from C57BL/6 mice were fused with B16 melanoma cells, and the resultant B16/DC hybrid cells expressed major histocompatibility complex (MHC) molecules - B7 as well as the B16 tumour marker M562 - which were enriched by Ia mediated positive selection with a MiniMACS column. The fusion rates were 12.7 26.8%. To generate hybrid tumour vaccines with potentially greater potent therapeutic efficacy, we genetically engineered DCs with granulocyte-macrophage colony-stimulating factor (GM-CSF) prior to cell fusion. Recombinant adenovirus vector was used to mediate gene transfer into DCs with high efficiency and DCs expressed GM-CSF at 96-138 ng/105 cells/ml 24 hr after GM-CSF gene transfer. GM CSF gene-modified DCs (DC.GM) exhibited higher expression of B7 and co stimulatory capacity in mixed lymphocyte reaction (MLR). Fusion of DC.GM with B16 cells generated B16/DC.GM hybrid cells secreting GM-CSF at 59-63 ng/105 cells/ml. Immunization of C57BL/6 mice with the B16/DC hybrid vaccine elicited a specific cytotoxic T-lymphocyte (CTL) response and protected the immunized mice from B16 tumour challenge, reduced pulmonary metastases and extended the survival of B16 tumour-bearing mice. The B16/DC.GM hybrid vaccine was able to induce a CTL response and protective immunity more potently and tended to be therapeutically more efficacious than the B16/DC vaccine. In vivo depletion of T-cell subsets demonstrated that both CD8+ and CD4+ T cells were essential for the therapeutic effects of B16/DC and B16/DC.GM hybrid vaccines. Additionally, other non-specific effector cells may also contribute to tumour rejection induced by the B16/DC.GM hybrid vaccine. These data indicate that a DC-based hybrid tumour vaccine may be an attractive strategy for cancer immunotherapy, and that GM-CSF gene-modified DCs may lead to the generation of hybrid vaccines with potentially increased therapeutic efficacy. PMID- 10457216 TI - Enhanced antigen-presenting activity and tumour necrosis factor-alpha-independent activation of dendritic cells following treatment with Mycobacterium bovis bacillus Calmette-Guerin. AB - Dendritic cells (DCs) are most potent among the antigen-presenting cells and are believed to be crucial for the initiation of a primary T-cell response to foreign antigens. Mycobacterial infection within macrophages is controlled by cell mediated immunity. To elucidate the stimulation of immune response by Mycobacterium bovis bacillus Calmette-Guerin (BCG), we purified DCs from precursor cells in human peripheral blood mononuclear cells (PBMC) by culturing them with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) and characterized their surface antigen expression. The interaction of cultured DCs with BCG resulted in increased surface expression of several DC-related marker antigens. BCG also induced reduction of endocytosis, enhancement of CD83 expression as well as B7 costimulatory molecules and IL-12 production, suggesting that BCG treatment directly induces DCs to mature. BCG treated DCs were much more potent antigen-presenting cells in allogeneic immune response than untreated DCs. Moreover, while the neutralization of tumour necrosis factor-alpha (TNF-alpha) significantly blocked the DC maturation induced by lipopolysaccharide (LPS), it could not inhibit the induction of DC maturation by the BCG treatment, indicating that TNF-alpha production plays a minor role in the BCG-induced DC maturation. However, the neutralization of TNF-alpha resulted in decreased IL-12 production by activated DCs. These results suggest that infection with BCG might evoke direct activation and maturation of DC and the general immune stimulant effect of BCG might be related with the activation of DCs. PMID- 10457217 TI - Glomerular overexpression and increased tyrosine phosphorylation of focal adhesion kinase p125FAK in lupus-prone MRL/MP-lpr/lpr mice. AB - Much progress has been made in understanding how mammalian cells receive a diverse array of external stimuli and convert them into intracellular biochemical signals. Such efforts have identified a large number of signalling molecules. However, our knowledge is limited as to their pathophysiological role in particular diseases. We demonstrate herein that an integrin-linked signalling molecule, focal adhesion kinase p125FAK (FAK), is overexpressed in glomeruli of lupus-prone MRL/MP-lpr/lpr (MRL-lpr) mouse as compared to its congeneic MRL-+/+ strain. Increased expression was specifically demonstrated in glomeruli but not in other tissues examined. The overexpression was observed in 16-week-old MRL-lpr mice with active nephritis, as well as in younger animals at 4 weeks of age. Thus, the upregulation of FAK clearly preceded the clinical onset of nephritis. FAK in MRL-lpr glomeruli is highly tyrosine phosphorylated and is associated with adapter protein Grb2. Previous in vitro studies have shown that the association of FAK/Grb2 links cell adhesion to the Ras pathway, which ultimately stimulates mitogen-activated protein (MAP) kinase, an important regulator of cell proliferation. In accordance, we observed constitutive MAP kinase activation in MRL-lpr glomeruli. Our findings suggest that signalling pathways involving FAK are activated in MRL-lpr glomeruli, and are likely to play a role in the development and progression of autoimmune-mediated murine nephritis. PMID- 10457218 TI - The human high-affinity immunoglobulin G receptor activates SH2-containing inositol phosphatase (SHIP). AB - On cytokine-primed U937 cells, aggregation of the human high-affinity immunoglobulin receptor, FcgammaRI, initiates signal transduction cascades which lead to the release of calcium from intracellular stores and no significant calcium influx. In these cells, aggregation of FcgammaRI results in no significant increase in inositol trisphosphate production, but rather phospholipase D is activated. Here we show that, in interferon-gamma (IFN-gamma) primed cells, the SH2 containing inositol 5' phosphatase, SHIP, is constitutively associated with the membrane fraction. Following aggregation of FcgammaRI, SHIP is rapidly and transiently tyrosine phosphorylated and becomes associated with the adapter molecule Shc. Shc also becomes tyrosine phosphorylated and translocates from the cytoplasm to the membrane fraction concomitant with the association between Shc and SHIP. Further, SHIP and Shc appear to be recruited to membrane-associated immune complexes following FcgammaRI aggregation. As no immunoreceptor inhibitory motif has been demonstrated to associate with FcgammaRI, these data suggest that SHIP may be recruited to the receptor through an SH2 domain interaction with Shc. PMID- 10457219 TI - Expression of B7.1 in a MUC1-expressing mouse mammary epithelial tumour cell line inhibits tumorigenicity but does not induce autoimmunity in MUC1 transgenic mice. AB - The MUC1 epithelial mucin, which is overexpressed and aberrantly glycosylated in breast and other carcinomas, is also expressed on the apical surface of most normal glandular epithelial cells. Since clinical trials evaluating the efficacy of MUC1-based vaccines have been initiated in breast cancer patients, it is important to address the question of whether an effective immune response to the cancer associated mucin can be generated without inducing autoimmunity. Since non classic cytotoxic T lymphocyte (CTL) responses to MUC1 have been reported, it is also relevant to examine the role of costimulatory molecules in the effective presentation of MUC1 based antigens. We have therefore looked at the effect of expressing B7.1 on the tumorigenicity of a MUC1 expressing mammary epithelial cell line (410.4) in a transgenic mouse expressing MUC1 on its normal glandular epithelial tissues. Coexpression of B7.1 with MUC1 in 410. 4 cells resulted in a dramatic inhibition of tumour growth which depended on the activity of CD4+ and CD8+ T cells. The epithelial tissues in the transgenic mice able to reject the B7.1, MUC1-expressing tumours showed no evidence of degeneration and the mice survived their normal life span. The results demonstrate that an immune response to the MUC1 antigen can be induced in MUC1 transgenic mice and that presentation of the antigen, whether directly or by cross-priming, is markedly enhanced by coexpression of B7.1. PMID- 10457220 TI - H-2Dd engagement of Ly49A leads directly to Ly49A phosphorylation and recruitment of SHP1. AB - We have used a number of in vitro and in vivo techniques to identify the molecules that can bind to the cytoplasmic tail of the Ly49A receptor. Affinity chromatography using peptides corresponding to the N-terminal 18 amino acids of Ly49A allowed the recovery of a number of proteins that bound preferentially to the tyrosine-phosphorylated peptide, including SH2-containing phosphatase-1 (SHP1) and the SH2-containing inositol 5' phosphatase (SHIP). In another approach, using the entire cytoplasmic domain of the Ly49A receptor, we found that SHP2 also interacted with the tyrosine-phosphorylated form of the Ly49A cytoplasmic tail. Using BIACORE(R)2000 analysis, we determined that both SHP1 and SHP2 bound to the tyrosine-phosphorylated cytoplasmic tail of Ly49A with affinities in the nanomolar range, whilst SHIP showed no binding. Mutation of tyrosine-36 to phenylalanine did not significantly affect the affinities of these proteins for the tyrosine-phosphorylated cytoplasmic tail of Ly49A. In addition, using a whole-cell system with T-cell lymphoma cell lines that expressed the Ly49A receptor or its H-2Dd ligand, we determined that engagement of Ly49A by its major histocompatibility complex (MHC) ligand leads to tyrosine-phosphorylation events and recruitment of SHP1. Recruitment of SHP1 was rapid and transient, reaching a maximum after 5 min. These data suggest that mechanisms for the inhibitory signal are generated following receptor engagement. They also provide direct evidence that ligand engagement of the Ly49A receptor is responsible for recruitment of downstream signalling molecules. PMID- 10457221 TI - Phenotypic identification of the subgroups of murine T-cell receptor alphabeta+ CD4+ CD8- thymocytes and its implication in the late stage of thymocyte development. AB - Phenotypic analysis of the medullary-type CD4+ CD8- [CD4 single-positive (SP)] thymocytes has revealed phenotypic heterogeneity within this cell population. The characteristic phenotype of mature peripheral T cells can be uniquely marked as Qa-2+ HSA- CD69-, whereas in the medullary-type CD4 SP thymocytes, the expression patterns of many markers were quite different. This suggests that there are many subgroups in the population, which reflects that medullary-type CD4 SP thymocytes may undergo phenotypic maturation. According to the results of two-colour flow cytometry, seven discrete phenotypes were identified by the expression capacity of Qa-2, HSA, CD69, 3G11 and 6C10 molecules. Consequently, the phenotypic precursor-progeny relationship can be envisaged as: 3G11- 6C10+ CD69+ HSAhi - >3G11+ 6C10+ CD69+ HSAhi --> 3G11+ 6C10- CD69+ HSAint -->3G11+ 6C10- CD69- HSAint Qa-2- -->3G11+ HSA-/lo Qa-2lo. At the stage of 3G11+ 6C10- CD69- HSAint Qa-2-, a branch pathway could be initiated, which gave rise to 3G11- HSAl degrees Qa-2- cells, which then, in turn, developed into 3G11- HSA-/loQa-2hi cells, a minor subgroup of the most mature CD4 SP cells. Consistent with this predicted pathway, experiments indicated that the first two subgroups were still cortisone sensitive, whereas the others were cortisone resistant. The cells in the last two Qa-2-positive subgroups are probably ready for emigration into the periphery. PMID- 10457222 TI - A bone marrow-derived stroma cell line, ST2, can support the differentiation of fetal thymocytes from the CD4+ CD8+ double negative to the CD4+ CD8+ double positive differentiation stage in vitro. AB - T-cell precursors differentiate into mature T cells predominantly in the thymus. However, it has also been reported that T-cell precursors mature in extrathymic organs such as the liver, bone marrow, or intestines. In order to investigate the nature of the extrathymic microenvironment that supports T-cell maturation, we examined the effect of a bone marrow-derived stroma cell line, ST2, on T-cell precursors by using a reaggregate thymic organ culture (RTOC) system. We found that ST2 cells supported the differentiation of fetal thymocytes at day 14.5 of gestation from a CD4- CD8- double negative (DN) to a CD4+ CD8+ double positive (DP) differentiation stage in a manner similar to that observed in thymus. Anti interleukin-7 receptor (IL-7R) and anti-c-kit antibodies blocked the growth of thymocytes in RTOC with ST2 cells, but did not inhibit the generation of DP thymocytes. These data indicate that a bone marrow-derived stroma cell, ST2, which supports B-cell differentiation, is also able to support T-cell development and may constitute one of the microenvironmental components for extrathymic T cell development. PMID- 10457223 TI - Integrin alpha1beta1 (VLA-1) mediates adhesion of activated intraepithelial lymphocytes to collagen. AB - Intraepithelial lymphocytes (IELs) from human intestinal epithelium are memory CD8+ T cells that bind to epithelial cells through human mycosal lymphocyte (HML) 1 and to mesenchymal cells through very late activation antigen-4 (VLA-4). Their binding of extracellular matrix proteins and the mechanism involved were tested. Activated 51Cr-labelled lymphocytes were incubated in protein-coated microwells with various additives. After washing, the adherent cells were detected by radioactivity. The percentages of activated IELs that bound to collagen types I and IV were 20 and 31%, respectively; fewer bound to fibronectin or laminin. Compared to interleukin-2-activated peripheral blood CD8+ T lymphocytes, more IELs bound collagen IV and fewer bound fibronectin. IEL adhesion to collagen (but not fibronectin or laminin) was up-regulated by antibody ligation of CD2 or by protein kinase C stimulation by phorbol ester; staurosporine reduced binding, while herbimycin, phytohaemagglutinin and CD3 ligation had no effect. Antibody blocking of integrin VLA-1 subunits alpha1 (CD49a) and beta1 (CD18) inhibited adhesion to collagen type I by 82+/-6% and to type IV by 94+/-1% (P<0.001), implicating VLA-1 as the main collagen receptor for IELs. Cell adhesion was dependent on extracellular divalent cations, a characteristic event of VLA-1 never before shown for IELs: manganese and magnesium ions supported binding in a dose-dependent manner; calcium ions inhibited their effectiveness. Therefore, IELs bind collagen through integrin alpha1beta1 after protein kinase C activation. Adhesion is modulated by divalent cations. PMID- 10457224 TI - Calcium-independent haemolysis via the lectin pathway of complement activation in the guinea-pig and other species*. AB - We previously reported that complement-dependent haemolysis of sheep erythrocytes (E) coated with mannan (M) and sensitized with human mannan-binding lectin (MBL) via the lectin pathway in man occurs in Mg-EGTA and requires alternative pathway amplification. Calcium was required for MBL binding to E-M, but once the E-M-MBL intermediate was formed, MBL was retained and haemolysis occurred in the absence of calcium. Comparable or greater lectin pathway haemolysis in the absence of calcium was observed upon incubation of E-M-MBL in guinea-pig, rat, dog and pig sera, and was further investigated in the guinea-pig, in which titres were much higher ( approximately 14-fold) than in man, and in contrast to humans, greater than classical pathway haemolytic activity. As in human serum, no lysis was observed in C4- or C2-deficient guinea-pig serum until purified C4 or C2, respectively, were restored. However, lectin pathway haemolytic activity in the guinea-pig did not require the alternative pathway. Removal (>98%) of factor D activity by three sequential passages through Sephadex G-75, resulting in serum which retained a normal classical pathway but no alternative pathway haemolytic activity, did not reduce the ability of guinea-pig serum to mediate haemolysis via the lectin pathway. Further, the C3-convertase formed via the lectin pathway (E-M-MBL-C4,2) lysed in C2-deficient guinea-pig but not human serum chelated with EDTA, a condition which precludes alternative pathway amplification. Thus, lectin pathway haemolysis occurs efficiently in guinea-pig serum, in the absence of calcium and without requirement for alternative pathway amplification. The guinea pig provides a model for studying the assembly and haemolytic function of a lectin pathway which contrasts with the lectin pathway of man, and allows for comparisons that may help clarify the role of this pathway in complement biology. PMID- 10457225 TI - Normal human immunoglobulin G4 is bispecific: it has two different antigen combining sites. AB - Unlike other immunoglobulin G (IgG) subclasses, IgG4 antibodies in plasma have been reported to be functionally monovalent. In this paper we show that the apparent monovalency of circulating IgG4 is caused by asymmetry of plasma IgG4. A large fraction of plasma IgG4 molecules have two different antigen-binding sites, resulting in bispecificity. Sera from patients with IgG4 antibodies to both house dust mite and grass pollen induced cross-linking of Sepharose-bound grass pollen antigen to radiolabelled house dust mite allergen Der p I. This bispecific binding activity was not observed in sera with IgG4 antibodies to either grass pollen or house dust mite exclusively. Depletion of IgG4 antibodies resulted in disappearance of the bispecific activity. By size exclusion chromatography we excluded the possibility that bispecific activity was caused by aggregation of IgG4 antibodies. These results indicate that circulating (polyclonal) IgG4 antibodies have two different antigen-binding sites and therefore are functionally monovalent antibodies. PMID- 10457226 TI - DNA activates human immune cells through a CpG sequence-dependent manner. AB - While bacterial DNA and cytosine-guanosine-dinucleotide-containing oligonucleotides (CpG ODN) are well described activators of murine immune cells, their effect on human cells is inconclusive. We investigated their properties on human peripheral blood mononuclear cells (PBMC) and subsets thereof, such as purified monocytes, T and B cells. Here we demonstrate that bacterial DNA and CpG ODN induce proliferation of B cells, while other subpopulations, such as monocytes and T cells, did not proliferate. PBMC mixed cell cultures, as well as purified monocytes, produced interleukin-6 (IL-6), IL-12 and tumour necrosis factor-alpha upon stimulation with bacterial DNA; however, only IL-6 and IL-12 secretion became induced upon CpG ODN stimulation. We conclude that monocytes, but not B or T cells, represent the prime source of cytokines. Monocytes up regulated expression of antigen-presenting, major histocompatibility complex class I and class II molecules in response to CpG DNA. In addition, both monocytes and B cells up-regulate costimulatory CD86 and CD40 molecules. The activation by CpG ODN depended on sequence motifs containing the core dinucleotide CG since destruction of the motif strongly reduced immunostimulatory potential. PMID- 10457228 TI - Editorial PMID- 10457227 TI - Mucosal immunogenicity and adjuvant activity of the recombinant A subunit of the Escherichia coli heat-labile enterotoxin. AB - The Escherichia coli heat-labile enterotoxin (LT) is an exceptionally effective mucosal immunogen and mucosal immunoadjuvant towards coadministered antigens. Although, in general, the molecular basis of these properties is poorly understood, both the toxic ADP-ribosylation activity of the LTA subunit and the cellular toxin receptor, ganglioside, GM1-binding properties of the LTB-pentamer have been suggested to be involved. In recent studies we found that GM1-binding is not essential for the adjuvanticity of LT, suggesting an important role for the LTA subunit in immune stimulation. We now describe the immunomodulatory properties of recombinant LTA molecules with or without ADP-ribosylation activity, LTA(His)10 and LTA-E112K(His)10, respectively. These molecules were expressed as fusion proteins with an N-terminal His-tag to allow simple purification on nickel-chelate columns. Their immunogenic and immunoadjuvant properties were assessed upon intranasal administration to mice, and antigen specific serum immunoglobulin-isotype and -subtype responses and mucosal secretory immunoglobulin A (IgA) responses were monitored using enzyme-linked immunosorbent assay. With respect to immunogenicity, both LTA(His)10 and LTA E112K(His)10 failed to induce antibody responses. On the other hand, immunization with both LT and the non-toxic LT-E112K mutant not only induced brisk LTB specific, but also LTA-specific serum and mucosal antibody responses. Therefore, we conclude that linkage of LTA to the LTB pentamer is essential for the induction of LTA-specific responses. With respect to adjuvanticity, both LTA(His)10 and LTA-E112K(His)10 were found to stimulate serum and mucosal antibody responses towards coadministered influenza subunit antigen. Remarkably, responses obtained with LTA(His)10 were comparable in both magnitude and serum immunoglobulin isotype and subtype distributions to those observed after coimmunization with LT, LT-E112K, or recombinant LTB. We conclude that LTA, by itself, can act as a potent adjuvant for intranasally administered antigens in a fashion independent of ADP-ribosylation activity and association with the LTB pentamer. PMID- 10457229 TI - Acute care nurses' experiences of moral certainty. AB - Moral certainty is a common, vivid experience for many nurses, but it may be a mixed blessing. From one perspective moral certainty provides comfort for the ethical decision-maker and may prompt necessary action which would have been impossible without it. From another perspective, moral certainty stifles dialogue and in-depth discussion of moral issues. Despite the fact that moral certainty is ubiquitous in health care, few research studies have explored both the positive and negative aspects of this complex and sometimes troubling concept. A qualitative design was used to describe the experiences of 20 acute care nurses with moral certainty. They were interviewed and the interviews analysed using Colaizzi's method for qualitative data analysis. These nurses' primary responses to moral certainty were 'speaking up', 'standing up' and 'refusing to participate'. Their experiences with moral certainty, responses to it, reasons for responding as they did, and clinical prompts are described. The author concluded that moral certainty may have positive and negative aspects. Making an effort to hear alternative views is suggested. PMID- 10457230 TI - Considering the nature of intersubjectivity within professional nursing. AB - The notion of intersubjectivity raises fundamental epistemological and ontological questions concerning how individuals come to know one another and how that knowing affects action. Within the sphere of professional nursing, relationship, as an intersubjective process between individuals, constitutes an integral element of professional nursing practice. Understanding the notion of relationship in terms of an intersubjective process is frequently laden with difficulty due to the polarization of intersubjectivity within either a traditional scientific position or a human science perspective. Reconciliation of both ways of knowing, however, may allow nurses to bring forward a broader base and scope of information to appreciate the experience of others. This article examines some of the notions of intersubjectivity and proposes an alternative understanding. PMID- 10457231 TI - Perspectives on surviving childhood sexual abuse. AB - This paper begins by examining sexual victimization and professional 'thinking sets' in relation to abuse. It goes on to offer an exploration of the literature in the area of childhood sexual abuse and its survival, and attempts to marshal some of this literature into naturally occurring conceptual groups with connecting theoretical threads. A summary of the theoretical paradigms which seem to produce dominant perspectives on the area is made. A cautionary discussion is introduced regarding the deterministic nature of the influence of these perceptual viewpoints upon research and therapeutic activity. Implications for nursing practice and education are drawn. PMID- 10457232 TI - Professional nurse autonomy: concept analysis and application to nursing education. AB - Professional nurse autonomy, an essential attribute of a discipline striving for full professional status, is often confused with personal autonomy, work autonomy or aggregate professional autonomy. Using Walker & Avant's (1995) model for concept analysis, this paper presents an analysis of professional nurse autonomy. Professional nurse autonomy is defined as belief in the centrality of the client when making responsible discretionary decisions, both independently and interdependently, that reflect advocacy for the client. Critical attributes include caring, affiliative relationships with clients, responsible discretionary decision making, collegial interdependence, and proactive advocacy for clients. Antecedents include educational and personal qualities that promote professional nurse autonomy. Accountability is the primary consequence of professional nurse autonomy. Associated feelings of empowerment link work autonomy and professional autonomy and lead to job satisfaction, commitment to the profession, and the professionalization of nursing. A student-centred, process-orientated curricular design provides an environment for learning professional nurse autonomy. To support the development of professional nurse autonomy, the curriculum must emphasize knowledge development, understanding, and clinical decision making. PMID- 10457233 TI - Meal procedures in institutions for elderly people: a theoretical interpretation. AB - In a study of mealtimes in institutions for elderly people the organization of meals was found to be task-oriented rather than patient-oriented in ways which failed to meet the needs of patients. The aim of the current paper is to examine and explain the institutional organization of meals, drawing on Goffmans' theory of institutionalized culture, Elias' theory of civilization, Douglas' theory of purity and order, and Bourdieus' key-concept 'habitus'. The method entails a secondary analysis of previous research carried out in a rehabilitation and long term hospital. This second analysis indicates that elderly patients coming to the ward with their individual meal customs were met by caregivers with an institutionalized culture. Meals in the ward were organized in line with the organization of family meals in society, and both the elderly people and the nurses strove towards civilized manners, purity and order. The demands implicit in these cultural practices kept the elderly patients silent. By failing into line, they suffered from their loss of habitus. In contrast nurses' habitus was accomplished by carrying out procedures automatically. Consequently, the combination of patients' lost habitus and nurses' working habitus resulted in defective nursing, where the purpose of nursing is the fulfilment of patients' social, personal and material needs. PMID- 10457234 TI - Developing realistic treatment standards in today's economic climate: stroke survivor education. AB - Teaching people about the risk factors and early symptom identification of stroke significantly decreases the incidence of both first time and recurrent strokes. Patient teaching has traditionally been a nursing responsibility. However, in the fiscal constraints of today's health care delivery system, nurses' time for teaching is highly restricted. This pilot study was designed to evaluate the impact of a short 1(1/2) hour stroke education programme in a population of hospitalized stroke survivors prior to their discharge. The first session focused on the disease process of stroke and its warning signs. A second session addressed risk factors and behavioural strategies for exercise, stress management, or smoking cessation. Responses to the teaching programme were evaluated through patient interviews immediately following the teaching session and 3 weeks later. Responses indicated that short-session patient teaching, in and of itself, is not always successful in changing behaviour but is successful for knowledge attainment. The findings from this pilot study suggests a need to consider new teaching strategies and a need to identify opportunities for reinforcement, support and encouragement that will meet the realities of today's time constraints while still meeting our professional responsibilities for patient education. PMID- 10457235 TI - The quality of life of cancer patients participating in phase I clinical trials using SEIQoL-DW. AB - The nature of phase I clinical trials in cancer patients raises many ethical issues. An area of major importance is the quality of life (QoL) of these patients. However, there is limited information available concerning the impact of phase I clinical trials on the QoL of patients with cancer. The primary aim of this study was to examine the QoL of cancer patients participating in phase I clinical trials. A descriptive triangulation approach was employed. The secondary aim was to determine the acceptability of the Schedule for the Evaluation of Individual Quality of Life - Direct Weighting (SEIQoL-DW) within this group of patients. SEIQoL-DW is a relatively new tool which investigates the areas identified by individual patients as important to their QoL. Cancer patients (n=15) participating in phase I clinical trials were interviewed using the SEIQoL DW tool. During this interview patients nominated the areas of life important to them and rated their relative weighting and current status. Cues nominated were described and results analysed using non-parametric statistical tests. The results showed that health and family were particularly important to patients in relation to their QoL. The importance of an individualized approach in the measurement of QoL was evident. SEIQoL-DW was found to be acceptable and practical to use in cancer patients participating in phase I clinical trials. Recommendations for practice were drawn and suggestions made for further research. PMID- 10457236 TI - Validation of the structure of the perceived meanings of cancer pain inventory. AB - This study examined the structure of a newly developed instrument measuring the meaning of cancer pain for Taiwanese patients. The Perceived Meanings of Cancer Pain Inventory (PMCPI) was developed based on Lazarus's cognitive theory of emotion and on findings from qualitative interviews. The 27-item PMCPI contains six scales: loss, threat, challenge, blame-others, blame-self, and spiritual awareness. Two hundred cancer patients from three hospitals in northern Taiwan who were experiencing pain were included in the study. A series of confirmatory factor analyses were conducted to test the fit of specified measurement models. The results indicated that only the loss, threat, challenge and spiritual awareness scales possessed good construct validity, while blame-other and blame self failed to show a satisfactory fit with the data. Correlation coefficients among the scales were also estimated. Parallel-item testing demonstrated that the items of each valid scale had the same factor loading and error variance. PMID- 10457237 TI - The psychometric properties of the diabetes management self-efficacy scale for patients with type 2 diabetes mellitus. AB - This article summarizes the development and validation of a scale to measure the level of self-efficacy of patients with type 2 diabetes mellitus. Self-efficacy is described as people's belief in their capability to organize and execute the course of action required to deal with prospective situations. This self-efficacy scale was developed based on the self-care activities these patients have to carry out in order to manage their diabetes. The following psychometric properties of this scale were established: content validity, construct validity, internal consistency and stability. The original scale contained 42 items. A panel of five experts in diabetes and four self-efficacy experts evaluated the original scale two times for relevance and clarity. This content validity procedure resulted in a final scale which consisted of 20 items. Subsequently, patients with type 2 diabetes were asked to complete this 20-item scale and further tests were done with the 94 usable responses. Factor analysis identified four factors, all of which were related to clusters of self-care activities used to manage diabetes which comprised this scale. The internal consistency of the total scale was alpha=0.81 and the test-retest reliability with a 5-week time interval was r=0.79 (P < 0.001). PMID- 10457238 TI - The critique of Heideggerian hermeneutical nursing research. AB - Within the past decade, over 25 research reports claiming to be based on Heideggerian interpretive phenomenology have been published in the nursing literature. This article reviews this research focusing on two critical issues. First, do the reports reflect a convergence of researcher understanding and participant narratives as called for by the Heideggerian tradition? Second, do Heideggerian ideas inform and enrich the studies' findings? The review reveals wide variations with regard to how these two issues are reflected in published reports. The author recommends that Heideggerian nurse researchers (a) seek to create a new narrative literature that allows for flexible and creative presentation of interpretive findings, while demanding adherence to sound interpretive scholarship, and (b) strengthen their partnerships with philosophers and other scholars so that ideas from Heideggerian interpretive phenomenology and other sources can not only guide their methods, but enrich their findings. PMID- 10457239 TI - Establishing the credibility of qualitative research findings: the plot thickens. AB - Qualitative research is increasingly recognized and valued and its unique place in nursing research is highlighted by many. Despite this, some nurse researchers continue to raise epistemological issues about the problems of objectivity and the validity of qualitative research findings. This paper explores the issues relating to the representativeness or credibility of qualitative research findings. It therefore critiques the existing distinct philosophical and methodological positions concerning the trustworthiness of qualitative research findings, which are described as follows: quantitative studies should be judged using the same criteria and terminology as quantitative studies; it is impossible, in a meaningful way, for any criteria to be used to judge qualitative studies; qualitative studies should be judged using criteria that are developed for and fit the qualitative paradigm; and the credibility of qualitative research findings could be established by testing out the emerging theory by means of conducting a deductive quantitative study. The authors conclude by providing some guidelines for establishing the credibility of qualitative research findings. PMID- 10457240 TI - Practice innovation: a methodological maze. AB - Practice innovation is an inevitable feature of a health culture preoccupied with evidence-based practice. The cyclical process of defining best practice, implementing and evaluating change represents an unparalleled opportunity for nurse researchers to engage in, and develop, practice through 'realistic evaluation'. However, the methodological dilemmas and challenges inherent in evaluation research which informs policy should not be under-estimated. This paper seeks to introduce and wrestle with some of the political tensions and methodological issues surrounding practice innovation when it is undertaken within an evaluative research framework. A critical pathway is presented to stimulate discussion and guide novice evaluators through this often perplexing methodological maze. A case study in audiological rehabilitation in elder care is used to illustrate the issues raised. PMID- 10457241 TI - Chinese elderly patients' perceptions of their rehabilitation needs following a stroke. AB - Stroke is the third leading cause of death and disability among Chinese elderly patients in Hong Kong and yet the rehabilitation needs of these patients are rarely explored. The aim of this study was to identify the rehabilitation needs of Chinese elderly patients following a stroke. The study adopted an ethnographic approach, information being gathered by the researcher through interviews with 15 key informants selected by purposive sampling. The perceptions of patients as to their own needs were sought at three stages of recovery - in the acute and rehabilitation settings and at 1 month following discharge. Ethical approval was gained from the Chinese University Faculty of Medicine ethical committee and access agreed by the hospital authorities. Verbal approval was gained from the patients before each interview, following confirmation of the voluntary nature of participation and assurance of confidentiality and anonymity. The researcher's role was also clearly stated. Analysis of the interview data produced five categories of patient need at the three stages of recovery, namely informational, physical, psychological, social and spiritual. The most frequently stated, but largely unmet, need in all settings was the need for information, particularly information about the reasons for stroke and about the activities that promote recovery. In the acute and rehabilitation settings patients' responses indicated a need to be respected as individuals, to be addressed by name and to be provided with privacy. Although the Barthel Index administered during interviews charted recovery at different rates, nurses did not always make links between the level of functional ability and the help needed with physical tasks. They also failed to recognize the relationship between physical and psychological needs and the equal importance of both in recovery from stroke. As Chinese elderly patients tend to take a passive role in seeking help and information, nurses play a significant role in the identification of individual rehabilitation needs. Implications for nursing practice are discussed. PMID- 10457242 TI - Recovering from stroke: a qualitative investigation of the role of goal setting in late stroke recovery. AB - This article examines the role of goal setting in the continuing relationship between specialist nurse and patients recovering from stroke. The nurse intervention was intended to ease the patient through the stages of recovery from stroke, focusing on emotional and social recovery rather than physical function. Literature on the use of goals in the nursing process is discussed. The article uses data from contemporary nurse records and from interviews with nurses and with patients and caregivers. The data from each of these elements were subjected to content analysis and were then synthesized using a grounded theory approach to interpret their significance. The perspective of patients and caregivers provides an additional insight into the use and limitations of goal setting which is largely developed in the literature from a nursing perspective. Nurses were found to have different interpretations of the use of goal setting. Some used it explicitly in their relationships with patients, whilst others used the concept to inform their actions whilst being less explicit and more informal. In all cases they demonstrate the tension between establishing and supporting progress towards realistic recovery goals and recognizing the limitations now placed on stroke victims. PMID- 10457243 TI - Health characteristics of elderly personal care home residents. AB - The purpose of this descriptive study was to explore the health characteristics of elderly personal care home residents. Personal care homes are community-based facilities that provide living arrangements, assistance with basic needs and protective oversight. Personal care home providers were interviewed regarding the health status of 80 elderly residents. The most frequent ADL needs were bathing, personal hygiene and dressing. Thirty-seven per cent of the residents had some bladder incontinence. Eighty-two per cent of residents needed some help with medication management. Over half of the residents had some cognitive impairment. These findings support the proposition of increased prevalence of cognitive impairment in personal care home residents as compared to the first studies a decade ago. The combination of functional deficits and cognitive impairment indicates that these elderly residents may have unmet health needs. The role of the registered nurse in this environment is discussed. PMID- 10457244 TI - Correlates of expected success at adherence to health regimen of people with IDDM. AB - Patient non-adherence is a well-recognized although poorly understood phenomenon that affects patients in all areas of health care. Failure to comply with health regimens is extremely costly both in economic terms and the health status of individuals. This study assesses factors which correlate with the expected success of health regimen adherence in 146 insulin-dependent diabetes mellitus (IDDM) subjects. The results indicate that success in complying with a health regimen is associated with good family support and rapport with health professionals, an absence of chronic stress and the capacity to take up the challenges posed by the disease. Health professionals have a role in engendering optimism, in maintaining enthusiasm, and facilitating and encouraging maintenance in health behaviours. PMID- 10457245 TI - Nurses' perceptions: issues that arise in caring for patients with diabetes. AB - Nurses' perceptions in caring for persons with diabetes have been little studied. To address this gap in the literature, a sample of nurses from a large Mid western health care system were surveyed on nurses' perceptions of: (i) problems encountered in the care of patients with diabetes; (ii) problems encountered by patients and/or family member(s) in diabetes management; and (iii) nurses' suggested solutions. A randomly selected list of 200 registered nurses obtained from the health system's Department of Human Resources included inpatient, outpatient, emergency department, medical centre and home health care nurses. The sample was stratified to include 25% inpatient and 75% outpatient nurses. Of the 200 surveys mailed, 136 were returned (68% response rate). Twenty-four per cent of the 136 nurses reported they did not provide care for patients with diabetes. Of 103 nurses providing care to patients with diabetes, 98% were female, 91% were Caucasian, 76% were between the ages of 30 and 49 years, 57% worked in outpatient settings, 35% worked in primary care, and 42% had a bachelor's degree or higher. Of those with practice guidelines, 84% found the practice guidelines helpful. These nurses also perceived that they, as nurses, needed more education to improve their care of diabetes patients; few nurses believed it was within the scope of their practice to change treatment regimens. The perception of most nurse respondents was that acceptance of diabetes, knowledge deficits and non compliance were primary patient problems in the management of diabetes. Nurses' perceptions of solutions to the problems centred on education of nurses and patients, and reinforcement of the importance of follow-up care. PMID- 10457246 TI - The relationship between diabetes knowledge and compliance among Chinese with non insulin dependent diabetes mellitus in Hong Kong. AB - This study examines the relationship between diabetes knowledge and compliance among Chinese with non-insulin dependent diabetes mellitus (NIDDM) in Hong Kong. A cross-sectional design was used to collect data through structured self-report interviews based on validated scales assessing diabetes knowledge, compliance behaviours and demographic data. The Diabetes Knowledge Scale was used to sample knowledge in the major areas of basic physiology of diabetes and general principles of diabetes care. Compliance level was assessed by using the Compliance Behaviour Questionnaire, inspection of patients' feet and the value of HbA1c. A convenience sample of 52 Chinese with NIDDM receiving out-patient diabetes care participated in the study on a voluntary basis. Descriptive and correlational statistical analyses were used to analyse the data. The findings indicated that there was no association between diabetes knowledge and compliance. There was a gap between what the patients were taught and what they were actually doing. Most of the patients gained higher marks on factual knowledge on diabetes but lost marks on the application of knowledge to their real life situations. Strategies are suggested to close the 'knowledge-action gap' and increase patients' motivation and ability to comply with the health regimen. PMID- 10457247 TI - Feelings of oncology patients about being nursed in protective isolation as a consequence of cancer chemotherapy treatment. AB - This qualitative research explores the feelings of oncology patients nursed in protective isolation following high-dose chemotherapy. Five patients described their feelings about the isolation experience during audio-taped interviews lasting between a half and one hour. Tapes were transcribed and analysed, with emergent themes considered in relation to the other interviews and to the literature. The research findings indicate that cancer patients have specific concerns with regard to their experience in the isolation environment, which fall into four distinct categories of: 'being shut in'; 'coping with the experience'; 'being alone'; and 'maintaining contact with the outside world'. In addition, patients have concerns with regard to the experience of having cancer, that impinge upon the isolation experience. These are: 'having cancer'; 'suffering chemotherapy'; 'knowing what to expect'; and 'developing relationships with the health professionals'. The core variable is 'something that I have to go through'. The nature of the relationship between the categories led to the development of an integrative model for exploring the feelings of cancer patients nursed in isolation. Most of the informants coped well with the isolation experience and described feeling supported by the nursing staff. Their overriding concern in fact, reflected a desire to receive information about their disease and reassurance regarding their treatment. Being in isolation appears to inhibit communication about these issues. PMID- 10457248 TI - The impact of seclusion on the treatment outcomes of psychotic in-patients. AB - The controversial practice of seclusion for the treatment of the acutely emotionally ill continues despite a lack of proven effectiveness. This retrospective cohort study examined the impact of seclusion alone and in combination with other important patient-centred treatment variables (medications), disease severity variables, psycho-social and demographic variables that best predicted outcome (as defined by length of hospital stay and reduced pathology at discharge) for a group of consecutively admitted psychotic in-patients. Comparisons of baseline and outcome measures for secluded and non secluded cohorts were made via t-test analysis, and patient-related variables were entered as groups for stepwise regression analysis to determine those factors that could best predict outcome. Data were collected via a comprehensive chart review of 85 patients with psychosis, who were consecutively admitted over an 8-month period to a psychiatric in-patient unit of a general community hospital in southern Ontario, Canada. Bigelow's Psychiatric Symptom Assessment Scale (PSAS) was applied to the chart admission and discharge data to determine both initial pathology scores and pathology change scores. Investigator-developed global rating scales measured the early pharmacological and psycho-social variables of interest. Although seclusion itself did not appear to be associated with any more or less improvement in mental status at discharge, seclusion was associated with a 12-day increase in length of hospital stay, raising additional questions about its therapeutic effectiveness. Early medication response together with seclusion predicted 16% of the total variance of length of hospital stay (P = 0.0011) while baseline pathology, being married and being employed together predicted 66% of reduced pathology at discharge (P = 0. 00001). Secluded subjects experienced an earlier age of illness onset, were employed or full-time students, exhibited poorer attitude towards hospital, were more likely to be substance or drug abusers, received greater doses of chlorpromazine-equivalent medications, and had longer lengths of hospital stay than their non-secluded cohorts. Two predictive regression equations were formulated on the basis of these results. This study has raised several questions for further prospective investigation, has validated previous findings in the literature, and has produced new findings that may be of interest to administrators and clinicians providing quality care to this high-risk population. PMID- 10457250 TI - Fatigue in chronic illness: the experience of individuals with chronic obstructive pulmonary disease and with asthma. AB - Although fatigue has been identified as a major problem for individuals with chronic obstructive pulmonary disease and with asthma, no research was found in which the symptom was directly studied in persons with these conditions. From studies carried out on various patient populations, it appears that fatigue has some specificity to disease state. Thus, it is important to describe the experience of fatigue within patient populations. To expand theoretical understanding of fatigue, qualitative research methods need to be applied. The purpose of this study therefore was to describe and compare the fatigue experiences of persons with chronic obstructive pulmonary disease (n=17) and with asthma (n=19). Data were obtained by use of a semi-structured questionnaire and were content analysed for categories and themes. There were many similarities between the fatigue experiences of the two groups. Fatigue is inextricably linked to laboured breathing. Although it interferes with their ability to carry out meaningful activities, the majority of individuals with chronic obstructive pulmonary disease or asthma cope well with it. The informants identified two types of coping strategies they use to manage their situation, which may be categorized as: problem-focused, including energy conservation, utilization and restoration; and emotion-focused, including being positive, accepting the physical limitations, distracting and normalizing. PMID- 10457249 TI - The effect of foot massage on patients' perception of care following laparoscopic sterilization as day case patients. AB - This randomized-controlled study examined the effects of foot massage on patients' perception of care received following surgery. The sample of 59 women who underwent laparoscopic sterilization as day case patients were randomly allocated into two groups. The experimental group received a foot massage and analgesia post-operatively, whilst the control group received only analgesia post operatively. Each participant was asked to complete a questionnaire on the day following surgery. This examined satisfaction, memory and analgesia taken. The 76% response rate was comparable with other patient satisfaction studies following day-case surgery. Statistical analysis showed no overall significant difference in the pain experienced by the two groups; however, the mean pain scores recorded following surgery showed a significantly different pattern over time, such that the experimental group consistently reported less pain following a foot massage than the control group. This study has attempted to explore the use of foot massage in a systematic way and is therefore a basis for further study. PMID- 10457251 TI - Myocardial infarction, spouses' reactions and their need of support. AB - The purpose of this study was to investigate the impact of myocardial infarction (MI) on survivor's spouses in terms of fears and symptoms during the patient's recovery period. In addition the researchers sought to evaluate the influence of the support by the health care professionals on spouses' adjustment. A total of 57 Finnish spouses participated in the study. Of the spouses 47 were female and 10 were male. Data were collected using a structured questionnaire distributed to the spouses at a rehabilitation session at 2 weeks-4 months after the MI. Topics covered included fears, emotional and physical symptoms and the spouses' experience of the sufficiency of the support that they had received from health care professionals during the patient's recovery. Data were analysed using quantitative methods including descriptive statistics and multivariate methods. Fears and symptoms were classified using factor analyses. For fears two factors emerged which were named: disease-related fears and personal fears. In respect of symptoms experienced by the spouse three factors emerged: one which describes emotional distress, one which describes dysfunction and one describing spouses' own vulnerability. The significantly most intensively experienced fears by the spouses were the disease-related fears followed by the personal fears. The most frequently reported symptom was dysfunction followed by emotional distress and vulnerability. In regression analysis emotional distress was predicted by personal fears, support from the health care professionals, the shock reaction, spouse's own health and time after MI. Dysfunction was predicted by spouse's own health and personal fears whereas vulnerability was predicted by spouse's own health and support from the health care professionals. Study findings show that in the planning of the care of patients with MI it is important to attend to the reactions and needs of spouses. Their resources are required for optimal rehabilitation of the patient. PMID- 10457252 TI - Synergy: working together with volunteers in community well woman clinics. AB - Despite promotion of the participation of volunteers in community and primary care services, there are few British studies which have explored the benefits and challenges of collaborative working between volunteer and paid workers, and in particular, women's experiences of volunteering within the National Health Service (NHS). The study presented here seeks to add to the limited, existing knowledge in this area. The experiences of volunteer and paid workers working together in two community well woman clinics are discussed, focusing on the benefits of working together for volunteers, paid workers and clients. Discussion is based on data collected over an 18-month period of participant observation in two community well woman clinics, interviews with 26 volunteer and paid workers and a review of the clinics' operational policy documents. Drawing on the concept of synergy, relationships between volunteer and paid workers are illuminated. In this paper it is argued that working together renders the opportunity for volunteer and paid workers to share insights, perspectives and approaches which enhance clients' health assessments. It provides an added value dimension to client care and yields benefits for the service provider. The implications of the study for practice and policy are considered. PMID- 10457253 TI - Individuals' perception of their quality of life following a liver transplant: an exploratory study. AB - This study explores individuals' perceptions of the effect of a liver transplant on their quality of life, focusing on the progression from dependence to independence physically, socially and psychologically. A phenomenological design using taped semi-structured interviews was used. The sample consisted of five patients attending the out-patients clinic at least 1 year following a liver transplant for chronic liver disease. The data were analysed using cluster analysis of transcribed interviews. Categories were identified as physical, social and psychological factors affecting their progression from dependence to independence pre- and post-transplant and specific factors were identified as significant in overcoming the stressors affecting this progression. The findings reflected Maslow's Hierarchy of Needs. Pre-transplant their physical problems prevented them fulfilling personal goals and addressing psychological issues, e.g. dying. Post-transplant any physical problems identified were insignificant to the participant. They were keen to socially integrate and be treated as normal; however, family and friends restricted their independence by continuing to 'wrap them in cotton wool' as they had before their transplant. Personality, incentives and Unit support were identified as imperative in their progression from dependence to independence. The findings demonstrate a need to impress on family and friends of patients following a liver transplant, their role in assisting the patient's progression from dependence to independence. PMID- 10457254 TI - The experience of women with primary biliary cirrhosis: a literature review. AB - Primary biliary cirrhosis (PBC) is a serious and life-threatening illness that mainly affects women. Epidemiological data on the prevalence of the illness are unclear. The experience of women with this chronic illness has not been explored within nursing research. A review of the literature concerning PBC therefore is based on general themes relating to chronic illness. A chronic illness has two meanings: the symbolic significance and the consequences for the individual. The symbolic significance of PBC can be related to symbolism relating to the liver in general and to the general assumption that liver disease is related to alcohol consumption. The consequences for the individual woman with PBC have been described as following a disease management trajectory. This may include appreciating the major symptoms of the illness. The main symptoms of PBC are fatigue and pruritus. These are both insidious and debilitating symptoms of unclear aetiology that can cause women with PBC problems when seeking an illness explanation. The symptoms may also interfere with the woman's body image and her caring role. It is suggested that the factors that relate to PBC may result in social isolation for women with the illness. PMID- 10457255 TI - Minimal interventions for problem drinkers: a review of the literature. AB - There is an increasing body of literature concerning the role in health promotion for nurses working in many health care settings. It has been argued that this role should include identifying those individuals whose life-style increases their risk of developing health problems, as well as providing appropriate advice and information. Life-style factors which may contribute to ill-health include problem drinking. This literature review presents a critique of studies of brief, or minimal, interventions for problem drinkers which have been conducted in both primary care and acute settings. The concept of minimal interventions is explored and the potential for nurses to assume a role in delivering such interventions is discussed. PMID- 10457256 TI - 'Coming out' as a life transition: homosexual identity formation and its implications for health care practice. AB - This paper explores the formation of homosexual identity through the complex transitional process of 'coming out'. Linear developmental models are discussed as is the influence of societal norms on identity formation. In considering the implications for professional practice the author challenges the heterosexism in health care and suggests ways in which health care professionals can provide a supportive environment for gay men and lesbians. PMID- 10457257 TI - Pseudohypoparathyroidism: from bedside to bench and back. PMID- 10457258 TI - Polycystic bone disease: A new, autosomal dominant disorder. AB - We describe a new heritable bone disease characterized radiographically by increasingly numerous and enlarging cyst-like lesions throughout the skeleton. Beginning in early childhood, a father, son, and daughter all suffered from progressively frequent pathological fractures involving such radiolucencies. Healing occurred uneventfully and with little residual pain or deformity. Biochemical parameters of mineral homeostasis and skeletal turnover were normal. Bone scanning showed increased radioisotope uptake primarily in fractures and in the largest collections of the lesions. The histopathology is uncertain, but may reflect a form of intraosseous lipomatosis. This unique condition, which we have provisionally named polycystic bone disease, is inherited as an autosomal dominant trait with a high degree of penetrance. PMID- 10457259 TI - Differential expression of human lysyl hydroxylase genes, lysine hydroxylation, and cross-linking of type I collagen during osteoblastic differentiation in vitro. AB - The pattern of lysyl hydroxylation in the nontriple helical domains of collagen is critical in determining the cross-linking pathways that are tissue specific. We hypothesized that the tissue specificity of type I collagen cross-linking is, in part, due to the differential expression of lysyl hydroxylase genes (Procollagen-lysine,2-oxyglutarate,5-dioxygenase 1, 2, and 3 [PLOD1, PLOD2, and PLOD3]). In this study, we have examined the expression patterns of these three genes during the course of in vitro differentiation of human osteoprogenitor cells (bone marrow stromal cells [BMSCs]) and normal skin fibroblasts (NSFs). In addition, using the medium and cell layer/matrix fractions in these cultures, lysine hydroxylation of type I collagen alpha chains and collagen cross-linking chemistries have been characterized. High levels of PLOD1 and PLOD3 genes were expressed in both BMSCs and NSFs, and the expression levels did not change in the course of differentiation. In contrast to the PLOD1 and PLOD3 genes, both cell types showed low PLOD2 gene expression in undifferentiated and early differentiated conditions. However, fully differentiated BMSCs, but not NSFs, exhibited a significantly elevated level (6-fold increase) of PLOD2 mRNA. This increase coincided with the onset of matrix mineralization and with the increase in lysyl hydroxylation in the nontriple helical domains of alpha chains of type I collagen molecule. Furthermore, the collagen cross-links that are derived from the nontriple helical hydroxylysine-aldehyde were found only in fully differentiated BMSC cultures. The data suggests that PLOD2 expression is associated with lysine hydroxylation in the nontriple helical domains of collagen and, thus, could be partially responsible for the tissue-specific collagen cross linking pattern. PMID- 10457260 TI - Parathyroid hormone [PTH(1-34)] and parathyroid hormone-related protein [PTHrP(1 34)] promote reversion of hypertrophic chondrocytes to a prehypertrophic proliferating phenotype and prevent terminal differentiation of osteoblast-like cells. AB - The effects of parathyroid hormone/parathyroid hormone-related protein (PTH/PTHrP) on late events in chondrocyte differentiation were investigated by a dual in vitro model where conditions of suspension versus adhesion culturing are permissive either for apoptosis or for the further differentiation of hypertrophic chondrocytes to osteoblast- like cells. Chick embryo hypertrophic chondrocytes maintained in suspension synthesized type II and type X collagen and organized their extracellular matrix, forming a tissue highly reminiscent of true cartilage, which eventually mineralized. The formation of mineralized cartilage was associated with the expression of alkaline phosphatase (ALP), arrest of cell growth, and apoptosis, as observed in growth plates in vivo. In this system, PTH/PTHrP was found to repress type X collagen synthesis, ALP expression, and cartilage matrix mineralization. Cell proliferation was resumed, whereas apoptosis was blocked. Hypertrophic chondrocytes cultured in adherent conditions in the presence of retinoic acid underwent further differentiation to osteoblast like cells (i.e., they resumed cell proliferation, switched to type I collagen synthesis, and produced a mineralizing bone-like matrix). In this system, PTH addition to culture completely inhibited the expression of ALP and matrix mineralization, whereas cell proliferation and expression of type I collagen were not affected. These data indicate that PTH/PTHrP inhibit both the mineralization of a cartilage-like matrix and apoptosis (mimicked in the suspension culture) and the production of a mineralizing bone-like matrix, characterizing further differentiation of hypertrophic chondrocytes to osteoblasts like cells (mimicked in adhesion culture). Treatment of chondrocyte cultures with PTH/PTHrP reverts cultured cells in states of differentiation earlier than hypertrophic chondrocytes (suspension), or earlier than mineralizing osteoblast-like cells (adhesion). However, withdrawal of hormonal stimulation redirects cells toward their distinct, microenvironment-dependent, terminal differentiation and fate. PMID- 10457261 TI - Adenovirus-mediated gene therapy of osteoblasts in vitro and in vivo. AB - Modulation of biological pathways governing osteogenesis may accelerate osseous regeneration and reduce the incidence of complications associated with fracture healing. Transforming growth factor beta1 (TGF-beta1) is a potent growth factor implicated in the regulation of osteogenesis and fracture repair. The use of recombinant proteins, however, has significant disadvantages and has limited the clinical utility of these molecules. Targeted gene therapy using adenovirus vectors is a technique that may circumvent difficulties associated with growth factor delivery. In this study, we investigate the efficacy of replication deficient adenoviruses containing the human TGF-beta1 and the bacterial lacZ genes in transfecting osteoblasts in vitro and osseous tissues in vivo. We demonstrate that adenovirus-mediated gene therapy efficiently transfects osteoblasts in vitro with the TGF-beta1 virus causing a marked up-regulation in TGF-beta1 mRNA expression even 7 days after transfection. Increased TGF-beta1 mRNA expression was efficiently translated into protein production and resulted in approximately a 46-fold increase in TGF-beta1 synthesis as compared with control cells (vehicle- or B-galactosidase-transfected). Moreover, virally produced TGF-beta1 was functionally active and regulated the expression of collagen IalphaI (5-fold increase) and the vascular endothelial growth factor (2.5-fold increase). Using an adenovirus vector encoding the Escherichia coli LacZ gene, we demonstrated that adenovirus-mediated gene transfer efficiently transfects osteoblasts and osteocytes in vivo and that transfection can be performed by a simple percutaneous injection. Finally, we show that delivery of the hTGF-beta1 gene to osseous tissues in vivo results in significant changes in the epiphyseal plate primarily as a result of increased thickness of the provisional calcification zone. PMID- 10457262 TI - Comparison of the effects of calcitonin gene-related peptide and amylin on osteoblasts. AB - Calcitonin gene-related peptide (CGRP) and amylin are homologous 37-amino-acid peptides which have been demonstrated to have anabolic effects on bone. It is not clear whether these effects are mediated by a common receptor, nor is it known which ligand is the more potent. These questions are addressed in the present study using cultures of fetal rat osteoblasts. CGRP increased cell number when present in a concentration >/=10-9 M, but 10-8 M CGRP was required to stimulate thymidine and phenylalanine incorporation. Amylin was effective on these indices at 100-fold lower concentrations, and its maximal effects were about twice as great as those of CGRP. ED50's for the effects of amylin and CGRP on cell number were 10-12 M and 10-10 M, respectively. There was no additivity between maximal doses of the peptides on these indices. The effects of specific receptor blockers on the maximal stimulation of cell number by these peptides were also studied. The CGRP receptor-blocker, CGRP-(8-37), completely blocked the effect of CGRP at blocker concentrations >/=10-9 M. In contrast, the amylin receptor blocker, amylin-(8-37), completely blocked the effects of CGRP when the blocker was present in concentrations as low as 10-11 M. The KI of CGRP-(8-37) was 2 x 10-10 M and that of amylin-(8-37) was 7 x 10-12 M. In converse experiments studying the blockade of maximal doses of amylin, amylin-(8-37) 10-10 M was effective (KI 1 x 10-10 M), whereas a 100-fold greater concentration of CGRP-(8-37) was necessary to achieve the same effect (KI 6 x 10-9 M). It is concluded that amylin and CGRP probably act through a common receptor to stimulate osteoblast growth, and that this receptor has a higher affinity for amylin than for CGRP. PMID- 10457263 TI - Failure to detect the extracellular calcium-sensing receptor (CasR) in human osteoblast cell lines. AB - Whether the known calcium-sensing receptor (CasR) is present in osteoblasts is a source of considerable controversy. Prior studies failed to detect CasR in osteoblasts, but more recent investigations purport the detection of CasR in several osteoblast cell lines by immunoblot analysis with polyclonal anti-CasR antisera (4637) and low stringency reverse transcriptase-polymerase chain reaction (RT-PCR). To explain these disparate findings, we performed immunoblot analysis with the 4637 anti-CasR antisera and a highly specific monoclonal antibody to CasR (ADD), and we compared the ability of low and high stringency RT PCR to amplify CasR transcripts. We found that the ADD antibody detected the anticipated CasR immunoreactive bands, including a approximately 165 kDa and approximately 140 kDa glycosylated doublet and a >250 kDa dimerized receptor, in positive control mouse kidney, human parathyroid, and human embryonic kidney (HEK) 293 cells transfected with rat CasR, but we did not detect these bands in either wild-type HEK 293 cells or Saos2, MG-63, or U-2 OS osteoblast-like cell lines. Standard two-step RT-PCR using CasR-specific primers confirmed these results by detecting CasR transcripts in positive controls but not in negative control HEK 293 cells or osteoblast cell lines. In contrast, the 4637 antisera did not recognize CasR by immunoblot analysis under the conditions studied and our low stringency RT-PCR procedure amplified nonspecific products in wild-type HEK 293 cells and osteoblasts. Since we failed to detect CasR in human osteoblast cell lines using either the highly specific ADD antibody or RT-PCR under standard conditions, it is possible that the cation response in osteoblasts is mediated by a functionally similar but molecularly distinct calcium sensing receptor. PMID- 10457264 TI - Modulation of osteoblast-like cell behavior by activation of protease-activated receptor-1. AB - In addition to playing a central role in thrombosis and hemostasis, the serine protease thrombin is a specific agonist for a variety of functional responses in cells including osteoblast-like cells. Many of the cellular responses to thrombin are mediated by protease-activated receptor-1 (PAR-1). Since osteoblasts express PAR-1 in vivo during development, the effect of PAR-1 activation on proliferation and differentiation in primary rat osteoblast-like cells was investigated. Thrombin or the rat PAR-1-activating peptide SFFLRNPSENTFELVPL (SFFL) stimulated cell proliferation (as assessed by 3H- thymidine incorporation) of primary osteoblast-like cells derived from long bone or calvaria, and treatment with antibodies to PAR-1 abolished the proliferative response to thrombin. Activation of PAR-1 by thrombin or SFFL inhibited endogenous alkaline phosphatase (ALP) activity and caused a transient elevation of intracellular calcium in the osteoblast-like cells. Calcium mobilization was not, however, required for thrombin's effect on proliferation or ALP activity. The ability of a number of growth factors and hormones to regulate expression of PAR-1 in osteoblast-like cells was investigated. Expression of PAR-1 transcript and protein by osteoblast like cells in vitro was markedly increased by treatment with transforming growth factor-beta (TGF-beta), and the proliferative response to thrombin was enhanced by TGF-beta pretreatment. Platelet-derived growth factor-BB caused a slight but significant down-regulation of PAR-1 mRNA expression. Thrombin caused a transient increase in PAR-1 expression, whereas neither parathyroid hormone-related peptide nor 1, 25-dihydroxyvitamin D3 had any effect. The observations described here suggest that PAR-1 mediates thrombin-induced osteoblast proliferation, which in turn may contribute to responses of osteoblasts to osteogenic growth factors. PMID- 10457266 TI - Polarity of osteoblasts and osteoblast-like UMR-108 cells. AB - Enveloped viruses, such as vesicular stomatitis virus (VSV) and Influenza virus, have been widely used in studying epithelial cell polarity. Viral particles of VSV-infected epithelial cells bud from the basolateral membrane, which is in contact with the internal milieu and the blood supply. Influenza-infected cells bud viral particles from the apical surface facing the external milieu. This feature can be utilized in labeling polarized membrane domains. We studied the polarity of mesenchymal osteoblasts using osteosarcoma cell line UMR-108 and endosteal osteoblasts in situ in bone tissue cultures. Immunofluorescence confocal microscopy revealed that the VSV glycoprotein (VSV G) was targeted to the culture medium-facing surface. In endosteal osteoblasts, VSV G protein was found in the surface facing bone marrow and circulation. On the contrary, Influenza virus hemagglutinin (HA) was localized to the bone substrate-facing surface of the UMR-108 cells. Electron microscopy showed that in the cases where the cells were growing as a single layer, VSV particles were budding from the culture medium-facing surface, whereas Influenza viruses budded from the bone substrate-facing surface. When the cells overlapped, this polarity was lost. Cell surface biotinylation revealed that 55% of VSV G protein was biotinylated, whereas Influenza virus HA was only 22% biotinylated. These findings suggest that osteoblasts are polarized at some point of their life cycle. The bone-attaching plasma membrane of osteoblasts is apical, and the circulation or bone marrow facing plasma membrane is basolateral in nature. PMID- 10457265 TI - The anti-androgen hydroxyflutamide and androgens inhibit interleukin-6 production by an androgen-responsive human osteoblastic cell line. AB - While androgens clearly have significant skeletal effects, the paracrine mediators of androgen action on bone are at present unclear. Interleukin-6 (IL-6) is a candidate cytokine that is produced by osteoblastic lineage cells and promotes osteoclastogenesis and bone resorption. Here, we assessed constitutive as well as IL-1beta- and tumor necrosis factor-alpha (TNF-alpha)-stimulated IL-6 mRNA expression by Northern analysis and protein secretion by immunoassay in a human androgen-responsive osteoblastic cell line (hFOB/AR-6) which contains approximately 4000 androgen receptors (ARs)/nucleus. Treatment with 5alpha dihydrotestosterone (DHT) dose-dependently inhibited constitutive and TNF alpha/IL-1beta-stimulated IL-6 mRNA steady-state levels in hFOB/AR-6 cells by 70 80% at 10-7 M. In addition, testosterone also suppressed TNF-alpha/IL-1beta stimulated IL-6 mRNA levels by 57%, while the adrenal androgen dehydroepiandrosterone had no effect. Of note, the specific AR antagonist, hydroxyflutamide, also inhibited IL-6 mRNA levels by 70%. Consistent with the Northern analyses, treatment with 5alpha-DHT, testosterone, and hydroxyflutamide also inhibited IL-6 protein production by 79%, 62%, and 71%, respectively (p < 0.001), while these agents had no effect on IL-6 soluble receptor levels. Finally, we demonstrated that hydroxyflutamide treatment of hFOB/AR-6 cells markedly inhibited the activation and binding of NF-kappaB (a known stimulator of IL-6 gene transcription) to its response element, thus providing a potential mechanism for its effect on IL-6 production by osteoblasts. These data are consistent with the hypothesis that suppression of osteoblast IL-6 production by androgens may mediate, at least in part, the antiresorptive effects of androgens on bone. Moreover, our findings also indicate that hydroxyflutamide, which is a known AR antagonist in most tissues, may function as a selective AR modulator for effects on IL-6 production by osteoblasts. PMID- 10457267 TI - Further characterization of cells expressing STRO-1 in cultures of adult human bone marrow stromal cells. AB - Primitive cells of the osteoblast lineage are not well characterized but are known to be present within the STRO-1+ fraction of adult human bone and marrow. A survey of human osteosarcoma cell lines revealed that STRO-1 is expressed by MG 63 but not SaOS-2. Among murine cell lines tested, expression of STRO-1 was detected in the bipotential (adipocyte/osteoblast) line BMS-2 but not the committed osteoblast precursor MC3T3-E1. A proportion of cultured adult human bone marrow stromal cells (BMSCs) consistently expressed the STRO-1 antigen. The expression of a range of cell surface antigens was studied in relation to STRO-1 by flow cytometry and several, including the bone/liver/kidney isoform of alkaline phosphatase (ALP), were found to subtype the STRO-1+ population of BMSCs. Further, BMSCs dual-labeled with antibodies recognizing STRO-1 and ALP could be assigned to one of four fractions: STRO-1-/ALP-, STRO-1+/ALP-, STRO 1+/ALP+, and STRO-1-/ALP+. Cells from each fraction could be isolated in high purity and, when recultured, remained viable and exhibited a limited degree of phenotypic stability. Using reverse transcriptase-polymerase chain reaction, cells in the four fractions were found to express different levels of transcripts for the parathyroid hormone receptor (PTHr) and bone sialoprotein (BSP). The expression of transcripts for the nuclear transcription factor core-binding factor alpha 1/osteoblast-specific factor-2 (CBFA1/OSF2) was restricted to those fractions expressing STRO-1 and/or ALP. Treatment with 10 nM dexamethasone consistently increased the proportion of cells present in those fractions which expressed the highest levels of transcripts for PTHr and BSP (STRO-1+/ALP+ and STRO-1-/ALP+) while simultaneously decreasing the proportion present in the STRO 1+/ALP- fraction. In conclusion, the expression of STRO-1 in vitro remains a characteristic of less well differentiated cells of the osteoblast lineage; in cultures of BMSCs and in established human osteosarcoma cell lines, there is an inverse association between the expression of STRO-1 and ALP; dual labeling of BMSCs with monoclonal antibodies recognizing STRO-1 and ALP permits the identification and isolation of cells of the osteoblast lineage at different stages of differentiation. PMID- 10457268 TI - Extracellular Ca2+ increases cytosolic free Ca2+ in freshly isolated rat odontoblasts. AB - Recent evidence suggests that extracellular Ca2+ may modulate cell function in mineralized tissue. To determine whether dentinogenic cells, in particular, are sensitive to extracellular Ca2+, fura-2 microfluorometry was used to monitor intracellular calcium levels in odontoblasts freshly isolated from rat incisor. In response to applications of 0.5-4.0 mM extracellular calcium (CaCl2), most odontoblasts (84%; 107/128) showed an increase in intracellular calcium. For the majority of these cells (70%; 75/107), the typical response was biphasic; there was an initial, transient increase in intracellular calcium which reached peak levels within 30-50 s and decayed rapidly, followed by a slower (> 300 s) recovery toward basal levels. In general, the response of these cells to calcium was repeatable and the mean calcium concentration for the half-maximal response was approximately 1.3 mM. This effect could be partially blocked by either 200 microM lanthanum, a nonspecific blocker of Ca2+ channels, or 20 microM dantrolene, a potent inhibitor of Ca2+ release from internal stores. Used in combination, lanthanum, and dantrolene nearly abolished the calcium response completely. In addition, this response was sensitive to the dihydropyridine sensitive calcium channel blocking agent nicardipine (60 microM), indicating a role for voltage-gated calcium channels during these events. These results show that odontoblasts respond to external calcium through mechanisms involving both influx of external calcium as well as release of calcium from internal stores and suggest a role for extracellular calcium in regulating the function of these cells. PMID- 10457269 TI - "Cell paralysis" as an intermediate stage in the programmed cell death of epiphyseal chondrocytes during development. AB - The efficient elimination of apoptotic cells depends on heterophagocytosis by other cells, which is difficult or impossible when the dying cells are embedded in an extracellular matrix. This situation is exemplified by the epiphyseal chondrocytes during the development of the chondroepiphyses of long bones. A detailed ultrastructural study identified an unusual type of epiphyseal chondrocyte which contained a very dark nucleus with irregular patches of condensed chromatin and a crenated nuclear membrane. The cytosol consisted of excessively expanded endoplasmic reticulum lumen, containing "islands" of cytoplasm and organelles. Since these cells appeared to be "in limbo," neither viable nor dead, they are referred to as "paralyzed" cells. By studying cells of intermediate morphologies, we were able to demonstrate the sequence of events leading to cell paralysis. It is proposed that the paralysis represents an intermediate state in the physiological cell death of epiphyseal chondrocytes in which destruction is orderly and avoids a inflammatory, potentially locally destructive, reaction. The cell is rendered paralyzed in terms of function but impotent in respect of damaging consequences. Paralysis is compared and contrasted with apoptosis, autophagocytosis, and necrosis and may represent another mode of programmed cell death in situations where cells are immature and/or where phagocytosis by neighboring cells is difficult. PMID- 10457270 TI - FokI polymorphism at translation initiation site of the vitamin D receptor gene predicts bone mineral density and vertebral fractures in postmenopausal Italian women. AB - A novel T/C polymorphism (ATG to ACG) at the translation initiation site of the vitamin D receptor (VDR) gene, defined by FokI restriction endonuclease, has been recently associated with variation in bone mineral density (BMD) and rates of bone loss in a group of postmenopausal Mexican-American women. The presence of the restriction site, designated as f, allows protein translation to initiate from the first ATG, while the allele lacking the site, indicated as F, initiates translation at a second ATG. In this study, we investigated the role of FokI polymorphism in a group of 400 postmenopausal women of Italian descent stratified for BMD into osteoporotic (n = 164), osteopenic (n = 117), and normal (n = 119) groups. There were 159 (41%) FF homozygotes, 55 (14%) ff homozygotes, and 186 (45%) Ff heterozygotes. In the whole population, we observed a weak association between FokI polymorphism and lumbar BMD (p = 0.06, analysis of covariance [ANCOVA]) but not with femoral neck BMD (p = 0.5, ANCOVA). Interestingly, the effect of FokI genotypes on lumbar BMD was influenced by the years since menopause such that differences in BMD related to different VDR allelic variants were greater among women in the first 5 years of menopause (p = 0.04, ANCOVA), progressively declining afterward. In addition, a significantly higher prevalence of ff genotype in osteoporotic than in osteopenic and normal women was observed (p = 0.04, Chi-square test). Finally, ff genotype resulted significantly over represented in the group of women with a vertebral fracture as compared with controls (p = 0.003, Chi-square test), equivalent to a relative risk of 2.58 (95% confidence intervals 1.36-4.91). We conclude that in this population, FokI polymorphism at the VDR gene locus accounts for a part of the heritable component of BMD at the lumbar spine. PMID- 10457271 TI - Alendronate increases bone density and bone strength at the distal radius in postmenopausal women. AB - In addition to the alendronate Osteoporosis Intervention Trial (FOSIT) core protocol 901-0A of 1908 enrolled patients, the use of peripheral quantitative computed tomography (pQCT) was explored for the assessment of response to therapy. Bone mineral and strength related parameters at two different sites at the distal radius were explored in a subset of the multicenter core study. One hundred and three patients were entered into the substudy and given either a daily dose of 10 mg of alendronate or placebo for 1 year. Measurements were done at months 0, 3, 6, and 12. Inclusion criteria were bone mineral density (BMD) measurements at the lumbar spine of -2 SD. The response to therapy was assessed by dual-energy X-ray absorptiometry in the lumbar spine and the hip, and by pQCT in the ultradistal and the shaft sites of the radius. In line with the FOSIT core study, alendronate increased BMD at the lumbar spine and the hip, and it decreased the serum biochemical markers of bone turnover. The substudy showed differences between the therapy and placebo group in trabecular bone density (8.4%, p = 0.095), in total density (6.8%, p = 0.009), and in the bone strength index (BSI) (15. 6 mm3, p = 0.037) at the ultradistal site due to treatment and no changes at the radius shaft. A significant correlation was observed between percentage changes from baseline in BMD of the lumbar spine, and in total density and bone strength at the ultradistal radius site in the treatment group, but not in the placebo group. Thus, the ultradistal radius site did respond to alendronate therapy. The increased bone density accompanied a significant gain in the BSI at the ultradistal site, a finding that might help explain the reduced wrist fractures in the alendronate Fracture Intervention Trial. PMID- 10457272 TI - Age- and gender-related differences in vertebral bone mass, density, and strength. AB - This study was designed to evaluate age- and gender-related differences in vertebral bone mass, density, and strength by dual-energy X-ray absorptiometry (DXA), quantitative computed tomography (QCT), peripheral QCT (pQCT), ash measurements, and biomechanical testing. The material comprised human lumbar vertebral bodies (L3) from 51 females and 50 males (age-range: 18-96 years). The results showed that females had significantly lower vertebral body bone mass (ash weight) than males at any given age. The decline in bone mass with age was parallel for females and males. The different bone density measurements cancellous ash density, total vertebral body ash density, DXA bone mineral density, QCT, and pQCT-showed no gender-related difference concerning numeric value or changes with age. Morphometrical measurements showed that females had smaller vertebral bodies (volumes) than males. Hence the females had significantly smaller cross-sectional area (CSA) of L3 than males (11.6 cm2 and 14.4 cm2, respectively). This led to females having lower maximum compressive load (N) than males at all ages, whereas maximum compressive stress (load/CSA) showed no gender-related difference. In conclusion, females have lower vertebral body bone mass than males at any given age, due to smaller vertebral bodies. Hence, maximum compressive load (strength not corrected for size) was lower in females. Vertebral body cancellous bone density and total-vertebral body density were equal when comparing genders, and no gender differences were found in the size-corrected strength: maximum compressive stress. The decrease with age in vertebral body compressive strength decrease was twice as large as the age decrease in density. PMID- 10457273 TI - Biochemical markers of bone turnover and prediction of hip bone loss in older women: the study of osteoporotic fractures. AB - To examine the ability of commercially available biochemical markers of bone formation and resorption to predict hip bone loss, we prospectively obtained serum and timed 2-h urine specimens from 295 women age 67 years or older who were not receiving estrogen replacement therapy. Serum was assayed for two markers of bone formation: osteocalcin (OC) and bone-specific alkaline phosphatase (BALP). Urine specimens were assayed for four markers of bone resorption: N-telopeptides (NTX), free pyridinolines (Pyr), free deoxypyridinoline (Dpyr), and C telopeptides (CTX). Measurements of hip bone mineral density were made at the time the samples were collected and then repeated an average of 3.8 years later. Higher levels of all four resorption markers were, on average, significantly associated with faster rates of bone loss at the total hip, but not at the femoral neck. Women with OC levels above the median had a significantly faster rate of bone loss than women with levels below the median, but there was no significant association between levels of BALP and hip bone loss. The sensitivity and specificity of higher marker levels for predicting rapid hip bone loss was limited, and there was considerable overlap in bone loss rates between women with high and low marker levels. We conclude that higher levels of urine NTX, CTX, Pyr, Dpyr, and serum OC are associated with faster bone loss at the hip in this population of elderly women not receiving estrogen replacement therapy, but these biochemical markers have limited value for predicting rapid hip bone loss in individuals. PMID- 10457275 TI - Urinary galactosyl-hydroxylysine in postmenopausal osteoporotic women: A potential marker of bone fragility. AB - Alterations of the collagen matrix, e.g., increased hydroxylation and glycosylation of lysyl residues in collagen I, were found in human osteoporotic bone, and it was suggested that they could alter the mechanical properties of skeleton. To test this hypothesis, we evaluated the content of galactosyl hydroxylysine (GHYL) in bone collagen, as assessed by its urinary excretion, and related it to the occurrence of fracture. Two hundred and fifteen unselected postmenopausal women with osteoporosis were divided in two subgroups (comparable for age, age of menopause, bone mineral density, and biochemical parameters of bone turnover) on the basis of the history of fragility fracture; 115 patients had suffered no fracture and 100 patients had suffered one or more fractures 3 or more years before. Four urinary markers of bone turnover (hydroxyproline, cross linked N-telopeptide, free deoxypyridoline, and GHYL) were evaluated in all patients. There was no difference between the two groups with regard to all the parameters studied except for GHYL, which was significantly higher in the group with a history of fracture (1.35 +/- 0.82 mmol/mol of creatinine [Cr] versus 1.03 +/- <0.48 mmol/mol Cr, p < 0.001); this marker did not correlate with other markers of bone remodeling in the fracture group, indicating a possible defect in bone collagen. In conclusion, provided that increased levels of urinary GHYL do reflect overglycosylation of hydroxylysine in bone collagen, the GHYL may be considered a marker of bone collagen quality. Our results, showing higher urinary GHYL in osteoporosis patients with fracture, seem to confirm this suggestion. PMID- 10457274 TI - Genetic markers, bone mineral density, and serum osteocalcin levels. AB - We evaluated five genetic markers for products that contribute to skeletal mineralization including the Sp1 polymorphism for type I collagen Ai (COLIA1), the vitamin D receptor (VDR) translation initiation site polymorphism, the promoter of the osteocalcin gene containing a C/T polymorphism, the estrogen receptor (ER) gene containing a TA repeat, and the polymorphic (AGC)n site in the androgen receptor. These markers were evaluated for their potential relationship with bone mineral density (BMD), measured by dual-energy X-ray densitometry, or its 3-year change. Additionally, potential associations of these genotypes and with baseline osteocalcin concentration or its 3-year change (assessed using radioimmunoassay) were evaluated. The study was conducted in 261 pre- and perimenopausal women of the Michigan Bone Health Study, a population-based longitudinal study of musculoskeletal characteristics and diseases. The polymorphic (AGC)n site in the androgen receptor showed a strong association with BMD of the femoral neck (FN) and lumbar spine and remained highly significant after adjusting for body mass index (BMI), oophorectomy/hysterectomy, oral contraceptive (OC) use and hormone replacement use (p < 0.001). The TA repeat at the 5' end of the ER gene was associated with total body calcium (p < 0.05) after adjusting for BMI, oophorectomy and hysterectomy, and OC use. The frequency of oophorectomy and hysterectomy within selected genotypes explained much of the statistically significant association of the ER genotypes with BMD of the FN and spine. There was no association of measures of BMD or bone turnover with the Sp1 polymorphism for COLIA1, the VDR translation initiation site polymorphism, or the C/T promoter polymorphism of the osteocalcin gene. These findings suggest that sex hormone genes may be important contributors to the variation in BMD among pre and perimenopausal women. PMID- 10457276 TI - Biological properties of salmon calcitonin IV. AB - In this study we characterized the biological activity of the recently identified salmon calcitonin (sCT) IV, in order to evaluate its potential therapeutic value. In the rat bioassay, sCT IV exhibited a 30% higher hypocalcemic activity than sCT I. The capacity of the molecule to inhibit bone resorption was assessed in vitro by the bone resorbing assay and the pit assay. An inhibitory effect, similar to that of sCT I, was observed in both assays. The interaction of sCT IV with the rabbit CT receptor was also studied. The affinity of sCT IV for the receptor was similar to that of sCT I, as was the potency for stimulating cAMP production. The antigenicity of the two molecules was not identical. Thus, this new CT could represent a useful novel therapeutic agent for the treatment of bone disorders. PMID- 10457277 TI - Functional characterization of two promoters in the human bone morphogenetic protein-4 gene. AB - Bone morphogenetic protein-4 (BMP-4) is a member of the BMP family, which consists of important regulators of bone formation and embryonic development. We have previously isolated the human BMP-4 encoding gene, which is associated with the heritable disorder Fibrodysplasia Ossificans Progressiva. In this study, we describe the molecular cloning and functional characterization of two promoters involved in the transcriptional regulation of the human BMP-4 gene, one upstream of exon 1, the second located in intron 1, upstream of exon 2. These two promoters give rise to different transcripts in a cell type- and differentiation dependent manner. Mutational analysis showed cell type-specific regulation of both promoter activities. Gel mobility shift assays indicated the presence of cell type-specific transcription factor binding sites in promoter 1. In addition, evidence was found for a novel BMP-4 transcript. Since various human diseases can be linked directly to aberrant expression of BMP genes, the present findings are of great importance in attempts to develop strategies for therapeutic interference with such diseases. PMID- 10457278 TI - Discordance between bone turnover and bone loss: effects of aging and ovariectomy in the rat. AB - Mechanical strain maintains bone architecture even under conditions of increased bone turnover such as occurs with ovarian hormone deficiency. The rat distal femur contains two sites that apparently experience different levels of mechanical strain and therefore the rat is a suitable model for investigating such effects. The femoral epiphysis experiences higher strain energy compared with the metaphysis and we report the effects of aging between 7 and 12 months and the postovariectomy effects over the same time period on cancellous bone variables measured at these two sites. Age-related bone loss in sham-operated (Sham) animals occurred in both regions, with a greater fall in the metaphysis than in the epiphysis (trabecular bone volume [BV/TV, %] Mean [SEM] Metaphysis: day 0, 25. 9 [2.4]; day 150, 8.8 [1.3]: Epiphysis: day 0, 44.8 [1.7]; day 150, 36.7 [1.4] [p < 0.0001]). With ovariectomy (OVX) there was a 73% reduction in cancellous bone at the metaphysis compared with no specific loss at the epiphysis (BV/TV [%] OVX: Metaphysis: day 150, 2.4 [0.4] [p < 0.01 compared with Sham]: Epiphysis: day 150 29.3 [2. 7] [NS]). Osteoblast cell activity and osteoclast surface were increased after ovariectomy in both regions. The mineral apposition rate decreased at 9.5 months of age in both regions (p < 0.0001), independent of ovariectomy, and was coincident with a reduction in trabecular number in the epiphyses of both operative groups and in the metaphysis of the ovary-intact group. These data suggest that local mechanical strain governs bone balance with aging and that architectural changes resulting from age-related bone loss may mirror those following estrogen deficiency but occur via a different cellular mechanism. PMID- 10457279 TI - Individual smallest detectable difference in bone mineral density measurements. AB - Bone mineral density (BMD) measurement is a major outcome measure in osteoporosis. The BMD changes observed must exceed the variability inherent in the measurement process to be considered related to disease progression. The objective of the study was to estimate short-term variability of BMD measurement and to propose a cut-off value for the smallest detectable BMD changes for an individual. To estimate the short-term variability, 70 healthy postmenopausal women aged 53 +/- 4 years (group 1) and 57 elderly osteoporotic postmenopausal women aged 80 +/- 6 years (group 2) had two repeated BMD measurements of the lumbar spine (L2-L4) and the proximal femur with dual-energy X-ray absorptiometry, with complete repositioning within 1 h. Cut-offs derived from short-term variability were either estimated from the coefficient of variation (CV) (which is a function of the measured value) or from the standard deviation (SD), and applied to 330 postmenopausal women (group 3) who had BMD measurements at baseline and 2 years later. The short-term intrasubject variability was greater at the lumbar spine in group 2 versus group 1 (0.0123 vs. 0.0059 g/cm2, p < 10-4), whereas it was not at the femoral neck (0.0098 vs. 0.0076 g/cm2, p = 0.28). There was no statistically significant correlation between short-term intrasubject variability (SD) and BMD as demonstrated with an analysis of covariance (p values ranging from 0.17 to 0.90). Cut-offs estimated with SD and CV were individually applied to group 3 patients. Using these two cut-offs, discrepancies in assessment of progression were observed in 1.7-8.6% of cases. Short-term BMD variability is constant in a wide range of BMD values. Consequently, to determine cut-off values for the smallest detectable differences in BMD at the individual level, precision errors should be based on SD (expressed in absolute units) rather than on CV (expressed in percentage). PMID- 10457280 TI - Food fractionation is a powerful tool to increase bone mass in growing rats and to decrease bone loss in aged rats: modulation of the effect by dietary phosphate. AB - The incidence of osteoporotic fractures has been associated with low bone mass. To reduce this incidence, it is therefore important to try to prevent the development of low bone mass by either increasing bone mass built up during adolescence and/or preventing bone loss in later life. It has been shown that food fractionation, a procedure that prevents the diurnal rhythm of bone resorption, increases bone mass in growing rats fed a high calcium (Ca), high phosphate (Pi) diet. In this paper, data are presented that show that providing growing rats with the same daily amount of a high Ca, low Pi diet (a Pi content similar to that of a human diet) in portions every 6 h instead of one meal increases total bone mineral content, trabecular bone mineral density, and cortical thickness, and markedly reduces the decrease in these parameters in aged rats. The effect is smaller when a high Ca, high Pi diet is fractionated. This could be the consequence of the transient postprandial increase in parathyroid hormone (PTH) following one large meal of a high Ca, high Pi diet, since the effect is similar to that reported after PTH injections which are anabolic for bone. Thus, a high dietary Pi must be considered as a confounding factor when treatments affecting bone mass are investigated in rats. The present data show that feeding habits have a profound effect on bone mass in the rat, independent of age. Whether bone mass in humans is also under the control of dietary habits is not yet clear. If so, frequent small meals of appropriate composition may help to prevent osteoporosis. PMID- 10457281 TI - Clinical vignette. Vitamin K plus vitamin D treatment of bone problems in a child with skeletal unloading. PMID- 10457282 TI - Is equivalent truly equivalent? PMID- 10457283 TI - It is not So simple PMID- 10457284 TI - Prader-Willi syndrome. AB - Prader-Willi syndrome is a multi system disorder characterized by neonatal hypotonia, later obesity, hyperphagia and mental retardation. It occurs sporadically, either as a result of microdeletion of chromosome 15p (70%) or as a result of maternal disomy of chromosome 15 (30%). The major problems encountered by parents are those of hyperphagia, food-seeking and obesity, and conduct disorder, particularly tantrums or oppositional behaviour. PMID- 10457285 TI - The diagnosis of Prader-Willi syndrome. AB - The methylation test can make the diagnosis of Prader-Willi syndrome (PWS) in approximately 99% of patients and is confirmed as a reliable, robust screening test. In a patient with PWS, methylation analysis does not provide the mechanism, for which other different genetic tests are required. Appropriate tests are available in each Australian state. PMID- 10457286 TI - Antibiotic therapy for Salmonella enteritis. PMID- 10457287 TI - Galactosaemia in Chinese. PMID- 10457288 TI - Varicella vaccine in non-immune household contacts of children with cancer or leukaemia. AB - OBJECTIVE: To assess the immunogenicity and reactogenicity of a new formulation of live attenuated varicella vaccine (Oka strain) in non-immune household contacts of children with cancer or leukaemia. METHODOLOGY: This was an open study with one group. Healthy varicella-susceptible adults and children living in the same household as children with cancer or leukaemia were vaccinated with a new live attenuated varicella vaccine (Oka strain) which is stable when stored at 2-8 degrees C (refrigerator temperature) for at least 24 months (Varilrix). Children less than 13 years of age received one dose (0.5 mL containing at least 103.3 plaque forming units) by subcutaneous injection and those aged over 13 years received two doses 8 weeks apart. Adverse reactions following vaccination were recorded daily by the vaccinees. Post-vaccination antibody estimation was determined using indirect immunofluorescence 6 weeks after vaccination. RESULTS: Thirty-five seronegative subjects (28 children and 7 adults and adolescents) were vaccinated. All subjects tested (34) had seroconverted after vaccination. Local injection site reactions were experienced by 15/35. Other adverse reactions were uncommon (rash 2/35, fever (>/= 37.5 degrees C) 3/35). No cases of clinical varicella occurred amongst the high-risk household contacts of the vaccine. CONCLUSION: This is the first study of this formulation of varicella vaccine in household contacts of children with cancer or leukaemia. The vaccine was found to be safe and immunogenic, but further follow-up is needed to document duration of immunity. PMID- 10457289 TI - Transverse cerebellar diameter on cranial ultrasound scan in preterm neonates in an Australian population. AB - OBJECTIVE: Fetal measurement of transverse cerebellar diameter (TCD) has been shown to correlate well with gestational age (GA), even in the presence of growth retardation. The aim of this study was to define the normal range of TCD in preterm neonates in an Australian population between 23 and 32 weeks GA. METHODOLOGY: Infants admitted to the Royal Women's Hospital, Melbourne, having routine cranial ultrasound scans (< 1500 g and/or of gestational age /= 3 ROP), 19 infants (12.1%) required cryo/laser therapy and one infant (0.6%) in this group had a retinal detachment. One hundred and fifty-seven of 159 surviving infants (98.7%) were examined for ROP. In the infants 27-28 weeks' gestation, 38.3% developed ROP (103 of 269 examined for ROP). Fifteen infants (5.6%) developed severe ROP, seven infants (2. 6%) required cryo/laser therapy for threshold ROP and three infants (1.1%) in this group had a retinal detachment. Two hundred and sixty-nine of 299 surviving infants (90%) were examined for ROP. In the infants 29-31 weeks' gestation, 10.8% developed ROP (48 of 443 examined for ROP). Six infants (1.4%) developed severe ROP, one infant (0.2%) required cryo/laser therapy for threshold ROP and no infant in this group had a retinal detachment. However, only 443 of 681 surviving infants (65.1%) in this group were examined for ROP. Of the four infants with detached retinas, one was a 25 week gestation infant weighing 840 g, two were 27 weeks' gestation weighing 960 and 980 g and one infant was a 28 week gestation infant weighing 620 g. No infant developed Stage 5 ROP. CONCLUSION: In the more mature infants 29-31 weeks' gestation, the rate of ROP is low, although severe ROP still occurs. However, only 65.1% of these infants were examined for ROP and we should be diligent in screening for ROP in the sicker infants in this group. The incidence of severe ROP as well as the rate of cryo/laser therapy in premature infants 23-26 weeks' in NSW has not changed since the increases seen in the early 1990s. Retinal detachment also occurs in the infants 27-28 weeks' gestation and it is important that all these infants are screened for ROP. PMID- 10457292 TI - Stress experienced by mothers of Malaysian children with mental retardation. AB - OBJECTIVE: To compare parenting stress among Malaysian mothers of children with mental retardation and a control group, and to determine factors associated with stress. METHODOLOGY: Seventy-five mothers of children with mental retardation aged 4-12 years and 75 controls (those without disabilities who attended the walk in paediatric clinic) participated in the Parenting Stress Index (PSI). Intelligence quotient (IQ) and Child Behaviour Checklist (CBCL) scores, together with sociodemographic data, were entered into a multiple stepwise regression analysis, using the PSI as the criterion. RESULTS: Mothers of children with mental retardation scored significantly higher than control subjects in both the child-related domain (difference between means 26.1, 95% confidence interval 19.6 32.5) and parent-related domain (difference between means 15.0, 95% confidence interval 7.9-22.1) of the PSI. The total child behaviour scores from the CBCL (P < 0.01), IQ scores (P < 0. 01) and sibship size (P < 0.01) were associated with child-related domain scores. For the parent-related domain, CBCL (P < 0.01) and IQ scores (P = 0.01) remained important factors but Chinese ethnicity (P < 0.01) and maternal unemployment (P < 0.01) were also significant predictors of stress. CONCLUSION: A large proportion of mothers of children with mental retardation experienced substantial parenting stress, especially Chinese and unemployed mothers, and this warrants appropriate intervention. PMID- 10457293 TI - Effect of nappy liners on temperature stability in very preterm infants. AB - OBJECTIVE: To determine whether absorbent liners used in posturally supportive cloth nappies influence temperature stability in infants < 31 weeks gestation. METHODOLOGY: Randomized cross-over trial conducted at King Edward Memorial Hospital for Women, Perth, Western Australia. Twenty-three infants nursed in incubators on Infant Servo Control were randomly assigned to wear cloth postural support nappies alternately with or without absorbent liners for 24-h periods over 4 days. Measurements of skin and incubator temperatures were recorded hourly. Times of all nappy changes and infant handling procedures were also recorded. RESULTS: There was no change in any temperature measurement over time, between days, or between day/night periods. Infants nursed with the liner demonstrated a higher skin temperature (0.04 degrees C), and a lower incubator temperature (1.05 degrees C). A drop in skin temperature of 0.02 degrees C and an increase in incubator temperature of 0.28 degrees C occurred following handling of infants. There was no effect due to sex, gestational age, or actual age of the infants. CONCLUSION: Use of an absorbent liner within a cloth postural support nappy promotes better temperature regulation in infants < 31 weeks gestation, by reducing incubator temperature and increasing skin temperature. PMID- 10457294 TI - Post-extubation prophylactic nasal continuous positive airway pressure in preterm infants: systematic review and meta-analysis. AB - OBJECTIVES: To determine whether management with nasal continuous positive airway pressure (NCPAP) in preterm infants having their endotracheal tube removed following a period of intermittent positive pressure ventilation (IPPV), leads to an increased proportion remaining free of additional ventilatory support, compared to extubation directly to headbox oxygen. METHODOLOGY: Search Strategy - Searches were made of the Oxford Database of Perinatal Trials, Medline, abstracts of conferences and symposia proceedings, expert informants, journal hand searching mainly in the English language and expert informant searches in the Japanese language. Selection criteria - All trials utilising random or quasi random patient allocation, in which NCPAP (delivered by any method) was compared with headbox oxygen for postextubation care were included. Methodological quality was assessed independently by the two authors. Data collection and analysis - Data were extracted independently by the two authors. Meta-analysis using event rate ratios (ERRs) and event rate differences (ERDs) was performed using Revman 3.0 statistical software. Prespecified subgroup analysis to determine the impact of different levels of NCPAP and use of aminophylline were also performed using the same package. Similar analysis to investigate the effect of postnatal age on outcomes of interest was also undertaken. RESULTS: Nasal CPAP, when applied to preterm infants being extubated following IPPV, reduces the incidence of adverse clinical events (apnoea, respiratory acidosis and increased oxygen requirements) indicating the need for additional ventilatory support. This result is both statistically significant and clinically important; ERR, 0.62 (0.49, 0.79) and ERD, - 0.175 (- 0.256, - 0.095). A trend towards reduction in the incidence of oxygen dependency at 28 days of age is also seen in the group extubated to NCPAP; ERR, 0.86 (0.67, 1.10) and ERD, - 0.069 (- 0.177, 0.039). CONCLUSION: Nasal CPAP is effective in preventing failure of extubation in preterm infants following a period of endotracheal intubation and IPPV. Further definition of the patient gestational age and weight groups to whom these results apply is required. Optimal levels of NCPAP as well as methods of administration remain to be determined. PMID- 10457295 TI - A clinical trial comparing oral azithromycin, cefixime and no antibiotics in the treatment of acute uncomplicated Salmonella enteritis in children. AB - OBJECTIVE: The objective of this study was to perform a prospective, randomized, controlled study to evaluate the role of azithromycin and cefixime in the treatment of uncomplicated non-typhoid Salmonella enteritis in children. METHODOLOGY: Patients with Salmonella enteritis were randomized to receive oral azithromycin (10 mg/kg/day once daily), cefixime (10 mg/kg/day divided twice daily) or no antibiotics for 5 days. The patients were followed up for the duration of their symptoms. Stool samples were sent for culture weekly following the therapy until two consecutive negative results were obtained. Susceptibility of the isolates to antibiotics was tested by the disk diffusion method. RESULTS: Forty-two patients with acute, uncomplicated, culture-confirmed Salmonella enteritis were studied. Duration of diarrhoea and time to defervescence after the therapy were not significantly different for patients treated with azithromycin, cefixime, or no antibiotics; there also were no significant differences with respect to the rate of clearance of Salmonella from stools among the three groups. Salmonella typhimurium was the most common serotype isolated. All 42 isolates were sensitive to cefixime, while two strains (5%) were resistant to azithromycin. CONCLUSION: Azithromycin or cefixime provides no benefit to paediatric patient with uncomplicated Salmonella enteritis. PMID- 10457296 TI - Transmission of tuberculosis from a seven-year-old child in a Sydney school. AB - OBJECTIVES: To determine whether a 7-year-old child with extrapulmonary and pulmonary tuberculosis (TB) and direct smear positive sputum for acid-fast bacilli was infectious to home and school contacts, and to ascertain potential adult sources of infection for these contacts. METHODS: Contact tracing by Mantoux testing was conducted on 220 children at a primary school and after school care facility, and 59 selected adults considered potential sources of infection. RESULTS: The participation rate for the children was 98% and 92% for the adults. Mantoux positivity (induration >/= 10 mm, or >/= 15 mm with previous BCG) among children was 13% at the school (anticipated rate 2-3%), 26% among school staff, and 7% among children at the after-school care centre where the index case attended. One exposed adult hospital staff member converted from Mantoux negative to positive. No other cases of TB disease were detected among children or adults tested. CONCLUSION: Although spread of TB from children to others is rare, the findings of this investigation indicate that transmission of TB from a young child to other children and an adult may have occurred, and that sputum testing and contact tracing for sputum smear positive children should be considered. PMID- 10457297 TI - Salmonella meningitis and its complications in infants. AB - OBJECTIVE: To review the presenting features, complications and outcome of infants with Salmonella meningitis. METHODOLOGY: Retrospective review of all cultures of cerebrospinal fluid positive for bacteria in children below 12 years of age, processed at the Department of Medical Microbiology, University of Malaya Medical Centre, Kuala Lumpur from 1973 to 1997. Records of all cases positive for Salmonella species were retrieved and studied. RESULTS: Thirteen infants aged 3 days to 9 months with Salmonella meningitis were included. The median age of onset of symptoms was 4 months. The clinical and laboratory features were similar to other causes of bacterial meningitis. Salmonella enteritidis was the commonest serotype isolated. Nine infants developed fits, six of which were difficult to control. Other complications noted were hydrocephalus (five), subdural effusions (four), empyema (three), ventriculitis (two), intracranial haemorrhage and cerebral abscess (one each). The use of ampicillin and/or chloramphenicol and inadequate duration of therapy resulted in recrudescence or relapse in five infants. The overall mortality was 18%. The presence of empyema, intracerebral abscess, ventriculitis, hydrocephalus, and intracranial haemorrhage were associated with adverse neurodevelopmental sequelae or death. More than half of those who survived had normal long-term outcome. CONCLUSION: Infants who developed neurological complications as a result of Salmonella meningitis had significant mortality and adverse long-term neurodevelopment outcome. PMID- 10457298 TI - Respiratory morbidity of hospitalized children with Trisomy 21. AB - OBJECTIVE: To review the respiratory morbidity in children with Trisomy 21 admitted to a teaching hospital. METHODOLOGY: A retrospective chart review of 232 admissions to John Hunter Children's Hospital during a 6.5-year period (1991-98). The primary outcome measures included: (i) primary reason for admission, (ii) concomitant respiratory pathology, (iii) admission to the Intensive Care Unit (ICU), (iv) length of stay and (v) costs of admission. RESULTS: Fifty-four per cent of admissions were primarily for respiratory tract pathology, dominated by pneumonia, bronchiolitis and croup. Admission to the ICU was required for 10% of admissions, most commonly for pneumonia (10/23). Congenital heart disease (CHD) was present in 33% of patients. The median length of stay and cost of admission for a child with Trisomy 21 (without CHD) with common respiratory conditions such as bronchiolitis, asthma or pneumonia was two to three times greater than in patients without Trisomy 21. CONCLUSION: Lower respiratory pathology is most common cause for acute hospital admission in children with Trisomy 21. The presence of CHD did not influence admission rates to hospital. However, patients with CHD had longer lengths of stay, appeared to have more severe illness, were more likely to require admission to an ICU and were more likely to require ventilatory support. PMID- 10457299 TI - The evolution of diagnostic trends in congenital heart disease: A population based study. AB - OBJECTIVE: To analyse historical trends in diagnosis of congenital heart disease (CHD) in all diagnosed cardiac malformations born between 1945 and 1994 in a population-based study, the first of its type. METHODOLOGY: Retrospective analysis of age and mode of diagnosis was carried out in 953 patients with CHD, in the setting of a regional hospital providing diagnostic and follow up services for all of Malta. Main outcome measures were age at diagnosis, cumulative percentage diagnosis by age and mode of diagnosis. RESULTS: A significant negative correlation of age at diagnosis with time was found for both lesions not requiring intervention, and requiring intervention (P < 0.0001). Cumulative percentage diagnosis by age increased progressively with time. Echocardiography increased the birth prevalence of definitively diagnosed defects, particularly of lesions not requiring intervention. The annual number of cardiac catheters for these conditions has decreased with an increased number and proportion of interventional catheters. The current catheterization rate for CHD is 6.1/100 000 population. CONCLUSION: Echocardiography has led to an increase in the diagnosis in the birth prevalence of CHD, and provided early diagnosis, supplanting cardiac catheterization as a primary diagnostic tool. PMID- 10457300 TI - Subglottic haemangioma: controversies in management. AB - OBJECTIVE: To discuss treatment modalities for subglottic haemangioma (SGH). METHODOLOGY: Case report of two children definitively managed by different modalities. RESULTS: Management by CO2 laser vaporization in one child, and laser followed by interferon 2-alpha in the second child were both successful in controlling the SGH without the need for tracheostomy. CONCLUSION: The stepped care approach at John Hunter Children's Hospital, Newcastle, New South Wales, is presented. Both laser surgery and interferon can help control SGH. Careful surveillance and interdisciplinary cooperation are essential to achieve a good outcome. PMID- 10457301 TI - Papilloedema secondary to acute purulent sinusitis. AB - Upper respiratory tract infections and sinusitis are common, but intracranial complications of sinusitis are rare in children and are often clinically unremarkable. Papilloedema secondary to purulent sinusitis is a complication previously not reported. We report two cases to highlight atypical manifestations of sinusitis in children. PMID- 10457302 TI - Classical galactosaemia in Chinese: A case report and review of disease incidence. AB - We report a case of galactose-1-phosphate uridyl transferase (GALT) deficiency in a full-term Chinese neonate, who presented with atypical biochemical features of hyperammonaemia in addition to the classical presenting features of jaundice and lethargy after feeding. Red cell GALT activity was virtually absent in the patient while 50% of normal activity was found in parents and a sibling. Mutation screening excluded both Q188R and N314D as the causative mutation in GALT gene, which suggested a possible genetic segregation among ethnic groups. Data from a Taiwan screening program suggested that the incidence of the disease was approximately 1 in 400 000 in the Chinese population which was a sixth of that in Caucasian populations. PMID- 10457303 TI - Deflazacort treatment in progressive diaphyseal dysplasia (Camurati-Engelmann disease). AB - Progressive diaphyseal dysplasia (PDD), a rare disorder of bones, in recent years has been accepted as a systemic disease within the spectrum of connective tissue disorders associated with immunological abnormalities. Steroids have been used in the treatment of PDD with variable success. In this report PDD is described in a 5-year-old boy who presented with leg pain, fatigue, headache and anorexia with an onset in infancy. Physical examination revealed a waddling gait, thorax deformity and thickening in the upper extremities. The diagnosis was made by radiologic demonstration of cortical thickening and a narrowed medullary cavity of the long bones of extremities. Bone scintigraphy showed areas of increased osteoblastic activity in the diaphyseal part of the long bones of extremities and the skull. Electron microscopic examination revealed myopathic and vascular changes. Serum immunoglobulin A, G and M levels were elevated and CD4 positive T cell numbers were low. Deflazacort, a steroid with a similar anti-inflammatory effect to prednisolone but with fewer adverse effects, was started in a dose of 1.2 mg/kg/day. Deflazacort treatment resulted in clinical and radiological improvement within 12 months with no side effects. In conclusion, steroids may be recommended as an effective method of treatment in PDD and deflazacort may be a safe alternative steroid. PMID- 10457304 TI - Radiofrequency catheter ablation in a haemodynamically compromised premature neonate with hydrops fetalis. AB - A preterm infant was born at 35 weeks gestation after failed antenatal antiarrhythmic therapy. The infant had an incessant supraventricular tachycardia, impaired ventricular function and hypotension and failed to respond to adenosine, cardioversion and intravenous amiodarone. After resuscitation from cardiovascular collapse, a successful radiofrequency catheter ablation (RFA) of a left free wall atrioventricular pathway was performed at 24 h of age without extracorporeal support. The infant is normal on follow up at 12 months of age. Whilst most fetal and neonatal supraventricular tachyarrhythmias respond to antiarrhythmic medications and RFA is not required, this is the earliest RFA to be performed on a premature infant when antiarrhythmics have failed. PMID- 10457305 TI - Childhood deaths and cargo barriers in cars. AB - To analyse features of childhood deaths associated with the absence or presence of car cargo barriers, three cases were taken from the files of the Department of Histopathology at the Women's and Children's Hospital and State Forensic Science Centre, Adelaide, Australia, over a 5-year period from January 1993 to December 1997. Case 1: A 6-year-old boy who was a rear passenger sustained fatal crush injuries from unrestrained luggage moving forward after an accident. No cargo barrier was in place. Cases 2 and 3: Two children aged 3 and 9 years, respectively, died from acute hyperthermia after they became trapped in the back of a station wagon with a fixed cargo barrier in place. Although car cargo barriers are excellent devices for preventing injury from unrestrained objects striking or crushing passengers, designing barriers that can be opened easily from the rear of the cabin would prevent children being trapped in potentially dangerous circumstances. PMID- 10457306 TI - Gentamicin doses and dose intervals in neonatal intensive care. PMID- 10457307 TI - Presentation and management of cystic neoplasms of the pancreas. AB - Pancreatic cystic neoplasms are uncommon, but it is important to differentiate them from pseudocysts and ductal adenocarcinoma. A retrospective review was performed to determine distinguishing characteristics and optimal treatment. In 51 patients operated on between 1981 and 1994 at a referral center, the following cystic neoplasms were found: 20 serous cystadenomas, 10 mucinous cystadenomas, 11 mucinous cystadenocarcinomas, five cases of mucinous ductal ectasia, and five papillary cystic neoplasms. Both mucinous ductal ectasia and papillary cystic neoplasms had distinguishing features when compared to other cystic neoplasms. Mucinous ductal ectasia was seen only in men, presented with typical symptoms, and had distinctive features on endoscopic retrograde cholangiopancreatography. Papillary cystic neoplasms occurred in young women (mean age 31 years) and were larger (mean 10.3 cm). Mucinous tumors were always symptomatic, whereas 55% of serous tumors were asymptomatic (P <0.001). The overall rate of resectability was 80%, and there was one operative death (2%). Intraoperative biopsy was diagnostic in 18 (78%) of 23 cases. An actuarial 5-year survival of 52% was found for resected mucinous cystadenocarcinomas. In conclusion, papillary cystic neoplasms and mucinous ductal ectasia have distinct characteristics that differentiate them from other types of pancreatic cystic tumors. Serous cystadenoma should be considered in asymptomatic patients and these patients should be closely observed. Symptomatic neoplasms should be resected with long-term survival expected for malignant forms. (J Gastrointest Surg 1998;2:504-508.) PMID- 10457308 TI - Role of antibiotics in acute pancreatitis: A meta-analysis. AB - In an attempt to decrease the infectious complications of acute pancreatitis and its high mortality, many investigators have conducted randomized prospective trials on the efficacy of prophylactic antibiotics. The results of these studies are conflicting, and many have called for a large multicenter study. Because multicenter trials are costly and difficult to organize, we believe that meta analysis is a reasonable alternative. A meta- analysis of all eight previously published trials of prophylactic antibiotics in acute pancreatitis was performed. The end point was death. The Mantel-Haenszel statistic was used to summarize odds ratios across studies in a fixed effects model, after homogeneity was assessed. Sensitivity analysis was performed as appropriate. The meta-analysis of all eight trials showed a positive benefit for antibiotics in reducing mortality. Sensitivity analysis showed that the advantage was limited to patients with severe pancreatitis who received broad- spectrum antibiotics that achieve therapeutic pancreatic tissue levels. It is recommended that all patients with severe pancreatitis be treated with broad- spectrum antibiotics that achieve therapeutic levels in pancreatic tissue. PMID- 10457309 TI - Is there a place for central pancreatectomy in pancreatic surgery? AB - Tumors located in the neck of the pancreas that are not small and superficial enough to be enucleated are usually resected with a pancreaticoduodenectomy or left splenopancreatectomy. Such operations may cause digestive disorders, glucose intolerance, and late postsplenectomy infection. Central pancreatectomy is a segmental resection whereby the cephalic stump is sutured and the distal stump anastomosed with a Roux-en-Y jejunal loop. The purpose of this study was to evaluate whether central pancreatectomy has a place in pancreatic surgery. Thirteen patients with the following tumors underwent central pancreatectomy: five endocrine tumors, one mucinous and six serous cystadenomas, and one solid cystic-papillary tumor. Mean operative time was 250 minutes. Operative mortality was zero. Complications occurred in three patients (23%). At mean follow-up of 68 months, no recurrences were found. Postoperative oral glucose tolerance, pancreolauryl, and fecal fat excretion tests were normal in all patients. We believe that central pancreatectomy does have a place in pancreatic surgery; it is a reliable technique for benign or low-grade malignant tumors and has a surgical risk similar to that of standard operations. Its principal advantage is that it preserves pancreatic parenchyma and the anatomy of the upper gastrointestinal and biliary tract and the spleen better than pancreaticoduodenectomy or distal pancreatic and splenic resection. (J Gastrointest Surg 1998;2:509-517.) PMID- 10457310 TI - Pancreatic polypeptide islet cell tumor: case report and review of the literature. AB - Pure pancreatic polypeptide-containing tumors (PPomas) are quite rare. Only 20 cases have been described. In this article we report a 75-year-old woman with such an endocrine islet cell tumor. The patient had no specific symptoms that could be ascribed to the tumor. An abdominal CT scan revealed a 3 cm soft tissue mass arising inferiorly from the tail of the pancreas. Local resection by way of a distal pancreatectomy was performed. A well-circumscribed hemorrhagic multiloculated mass, 3.7 cm in greatest dimension, was present in the tail of the pancreas. The patient has remained well and tumor free for the past 22 months. The endocrine characterization of the tumor was achieved by means of immunohistochemical analysis. Staining specific for insulin, glucagon, somatostatin, and gastrin was negative. In contrast, staining of the tumor for pancreatic polypeptide was strongly positive. A number of nonfunctioning islet cell tumors of the pancreas have been described. The lack of function has previously been suggested to indicate the lack of secretion of an endocrine product. This report documents that islet cell tumors may function by secreting pancreatic polypeptide but not cause symptoms. PMID- 10457311 TI - Cystic glucagonoma: A rare variant of an uncommon neuroendocrine pancreas tumor. AB - Glucagon-producing neuroendocrine tumors typically present with a characteristic constellation of symptoms including necrolytic migratory erythema, non-insulin dependent diabetes, weight loss, anemia, glossitis, and an increased thrombotic tendency. Most glucagonomas are solid and arise in the body or tail of the pancreas. We report two cases of cystic glucagonoma, one found incidentally in an asymptomatic patient and one in a patient with weight loss and diabetes but no rash. In the first patient, distal pancreatectomy and splenectomy were curative, whereas the second patient continued to exhibit elevated serum glucagon levels and symptoms of glucose intolerance in the absence of demonstrable metastases. Cystic glucagonoma is a unique variant of classic glucagonoma and should be considered in the differential diagnosis of cystic pancreatic neoplasms. PMID- 10457312 TI - Mucosal production of complement C3 and serum amyloid A is differentially regulated in different parts of the gastrointestinal tract during endotoxemia in mice. AB - The effect of endotoxemia and sepsis on mucosal production of the acute-phase proteins complement component C3 and serum amyloid A (SAA) was studied in mice. In addition, the role of the proinflammatory cytokines tumor necrosis factor alpha, interleukin (IL)(-1)beta, and IL-6 on mucosal C3 and SAA production was examined. Endotoxemia was induced by the subcutaneous injection of 250 microg/mouse of lipopolysaccharide. Control mice were injected with corresponding volumes of sterile saline solution. Sepsis was induced by cecal ligation and puncture, and sham-operated mice served as controls. Endotoxemia resulted in increased mucosal C3 levels in all parts of the gastrointestinal tract examined, from the stomach to the colon, with the most pronounced effects noticed in the proximal gastrointestinal tract. The influence of endotoxemia on mucosal SAA production was more differentiated with increased levels noted in the jejunum and ileum, and no changes seen in gastric and colonic mucosa. Sepsis resulted in similar changes in mucosal C3 and SAA levels as seen in endotoxemic mice, except that SAA levels were increased in colonic mucosa of septic mice. Among the cytokines, IL(-1)beta resulted in the most pronounced changes in mucosal acute phase proteins. The increase in C3 and SAA levels in the mucosa of the small intestine during endotoxemia was partially blocked by IL(-1) receptor antagonist. The results suggest that endotoxemia is associated with increased mucosal C3 production in different parts of the gastrointestinal tract and increased SAA production in the mucosa of the small intestine. Mucosal acute-phase protein synthesis may, at least in part, be regulated by IL(-1)beta. PMID- 10457314 TI - Comparison of thoracoscopic and laparoscopic Heller myotomy for achalasia. AB - For more than three decades experts have debated the relative merits of thoracoscopic Heller myotomy (no antireflux procedure) vs. laparoscopic Heller myotomy plus Dor fundoplication for treatment of achalasia. The aim of this study was to compare the results of these two methods with respect to (1) relief of dysphagia, (2) incidence of postoperative gastroesophageal reflux, and (3) hospital course. Sixty patients with esophageal achalasia were operated on between 1991 and 1996. Thirty underwent a thoracoscopic Heller myotomy and 30 had a laparoscopic Heller myotomy with a Dor fundoplication. The two groups were similar with respect to demographic characteristics, clinical findings, and extent of manometric abnormalities. Preoperative pH monitoring showed abnormal reflux in two patients in the laparoscopic group. Average hospital stay was 84 hours for the thoracoscopic group and 42 hours for the laparoscopic group. Excellent (no dysphagia) or good (dysphagia less than once a week) results were obtained in 87% of patients in the thoracoscopic group and in 90% of patients in the laparoscopic group. Postoperative pH monitoring showed abnormal reflux in 6 (60%) of 10 patients in the thoracoscopic group and in 1 (10%) of 10 patients in the laparoscopic group. The two patients in the laparoscopic group who had reflux preoperatively had normal reflux scores postoperatively. Laparoscopic Heller myotomy with Dor fundoplication was found to be superior to thoracoscopic Heller myotomy. Both operations relieved dysphagia, but the laparoscopic approach avoided postoperative reflux and even corrected reflux present preoperatively. In addition, the patients were more comfortable and left the hospital earlier following a laparoscopic myotomy. Whether it is truly possible to perform a Heller myotomy without an antireflux procedure in a way that relieves dysphagia and regularly avoids reflux remains questionable. PMID- 10457313 TI - Gastroesophageal reflux disease and mucosal injury with emphasis on short-segment Barrett's esophagus and duodenogastroesophageal reflux. AB - Gastroeosphageal reflux disease has been associated with long segments of Barrett's esophagus 6 mm Hg), overall length (normal >2 cm), and abdominal length (normal >1 cm), and their outcomes were assessed. Each group was subsequently divided into patients presenting with a primary symptom that was "typical" (heartburn, regurgitation, or dysphagia) or "atypical" (gastric, respiratory, or chest pain) of gastro-esophageal reflux, and outcome was assessed. Median duration of follow up was 18 months after surgery. Overall, laparoscopic fundoplication was successful in relieving symptoms of gastroesophageal reflux in 90% of patients. Patients with a typical primary symptom had an excellent outcome irrespective of the resting status of the LES (95% and 97%, respectively). Atypical primary symptoms were significantly more common in patients with a normal LES (29%) than in those with a structurally defective LES (10%; P <0.05), and these symptoms were less likely (50%) to be relieved by antireflux surgery. Laparoscopic antireflux surgery is highly successful and not dependent on the status of the resting LES in patients with increased esophageal acid exposure and primary symptoms "typical" of gastroesophageal reflux. Antireflux surgery should be applied cautiously in patients with atypical primary symptoms. PMID- 10457316 TI - Magnetic resonance cholangiopancreatography accurately predicts the presence or absence of choledocholithiasis. AB - Accurate common bile duct (CBD) imaging in patients with biliary calculi is an important determinant of specific therapy. Noninvasive methods to evaluate calculi in the CBD have limited accuracy and rely mainly on ultrasonography and computed tomography. Magnetic resonance cholangiopancreatography (MRCP) is a new noninvasive modality available to evaluate the biliary system. This study was undertaken to assess the accuracy of MRCP in predicting the presence or absence of CBD stones in patients at increased risk for choledocholithiasis. The medical records of 48 patients with a final diagnosis of biliary calculous disease undergoing MRCP between November 1995 and April 1997 were retrospectively reviewed. Three groups were identified: choledocholithiasis (n = 19), gallstone pancreatitis (n 5 11), and uncomplicated cholelithiasis (n = 18). In all patients the presence or absence of CBD calculi, as determined by MRCP, was correlated with the final diagnosis obtained from endoscopic retrograde cholangiopancreatography (ERCP) (n = 19), intraoperative cholangiography (n = 6), CBD exploration (n = 13), or clinical follow-up (n = 10). Sensitivity, specificity, and accuracy of MRCP were determined. The major clinical indications for MRCP in the 48 patients ware abnormal liver function tests followed by hyperamylasemia. Twenty patients were diagnosed with CBD stones and 28 were not. MRCP correctly predicted the presence of CBD stones in 19 of 20 patients and failed to detect CBD stones in one patient with gallstone pancreatitis. MRCP incorrectly predicted the presence of CBD stones in 3 of 28 patients ultimately found to have gallstones and no CBD stones. MRCP correctly predicted the absence of CBD stones in the other 25 patients including 10 patients with gallstone pancreatitis. Overall, MRCP had a sensitivity of 95%, a specificity of 89%, and an accuracy of 92%. MRCP is an accurate, noninvasive test for evaluating the CBD duct for the presence or absence of calculi in patients suspected of having CBD stones. Our data support the use of MRCP in the preoperative evaluation of these patients as findings may influence therapeutic decisions. PMID- 10457317 TI - Detection of aerosolized cells during carbon dioxide laparoscopy. AB - Laparoscopic surgery for malignancy has been complicated by port-site recurrences. The exact mechanism has yet to be defined. In vitro studies suggest that carbon dioxide-induced tumor cell aerosolization may play a role. We have attempted to document this in a human model. Patients scheduled for elective laparoscopy underwent port placement and abdominal insufflation with carbon dioxide. A suction trap was then filled with 40 cc of normal saline solution and attached to an insufflation site on the port. The carbon dioxide effluent was directed through the saline. The specimen was concentrated, resuspended, and transferred to a slide. A Papanicolaou stain was used. Thirty-five specimens were obtained. Fifteen patients (37%) had malignant disease, which was metastatic in eight. Five patients had carcinomatosis. In two of those with carcinomatosis, staining revealed a large number of malignant cells. Malignant cells were not found in any other patients. In two patients, however, aerosolized mesothelial cells were identified. Follow-up ranged from 2 to 7 months. One patient who displayed cellular aerosolization developed a port-site recurrence. We conclude that malignant cells are aerosolized but only during laparoscopy in the presence of carcinomatosis. It is unlikely that tumor cell aerosolization contributes significantly to port-site metastasis. PMID- 10457318 TI - Small-diameter prosthetic H-graft portacaval shunt: definitive therapy for variceal bleeding. AB - Partial portal decompression has become a popular option in the treatment of complicated portal hypertension. This study was undertaken to report long-term follow-up after partial portal decompression obtained utilizing 8 mm prosthetic H graft portacaval shunts. A total of 110 consecutive patients underwent H-graft portacaval shunting through a protocol that detailed care and studies from 1988 to 1996. Prospective follow-up recorded efficacy of partial portal decompression, shunt patency, morbidity of shunting, and survival. Seventy males and 40 females, whose average age was 54 +/-12.7 years (standard deviation), underwent shunting. Cirrhosis was due to alcohol abuse in 64%. Fourteen percent were in Child's class A, 55% in Child's class B, and 31% in Child's class C. Shunts were undertaken as emergencies in 20%, urgently in 13%, and electively in 67%. Shunting decreased portal pressure in all patients (30 +/-5.3 Hg to 19.9 -/+5.5 mm Hg; P <0.001). Early and late thrombosis was 6.4% and 3.6%, respectively. Late rebleeding occurred in 5.4%. Perioperative (30-day) mortality was 11.8%, and was highest for patients in Child's class C. Three-year survival was 53%. Five-year survival was 41%. Partial portal decompression is achieved with H-graft portacaval shunting. Rebleeding, shunt occlusion, and encephalopathy are uncommon. In this series of unselected older patients with alcoholic cirrhosis, 5-year survival after H-graft portacaval shunting was greater than 40% with minimal intervention. PMID- 10457319 TI - Near-total completion gastrectomy for severe postvagotomy gastric stasis: analysis of early and long-term results in 62 patients. AB - The aim of this study was to evaluate results of completion gastrectomy for severe postgastrectomy gastric stasis. A total of 51 women and 11 men underwent completion gastrectomy for gastric stasis between 1985 and 1996; follow-up was complete in 98% at 5.4 +/- 5 years. All patients had modified Visick scores preoperatively of grade III (37%) or IV (63%). Presentation included combinations of nausea, vomiting, postprandial pain, chronic abdominal pain, and chronic narcotic use. All had undergone prior vagotomy and had a median of four previous gastric operations. Hospital mortality was zero. Complications occurred in 25 patients (40%) and included the following: narcotic withdrawal syndrome (18%), ileus (10%), wound infection (5%), intestinal obstruction (2%), and anastomotic leak (5%). All or most symptoms were relieved in 43% (Visick grade I or II), but 57% of the patients remained in Visick grade III or IV. Nausea, vomiting, and postprandial pain were reduced from 93% to 50%, 79% to 30%, and 58% to 30%, respectively (P<0.05), but chronic pain, diarrhea, and dumping syndrome were not significantly affected. Univariate analysis revealed no preoperative characteristic to be predictive of good outcome. Logistic regression analysis suggested that the combination of nausea, need for total parenteral nutrition, and retained food in the stomach predicted a poor outcome (P<0.05). Completion gastrectomy is successful in 43% of patients. The combination of nausea, need for total parenteral nutrition, and retained food at endoscopy are negative prognostic factors. PMID- 10457320 TI - Effect of microscopic resection line disease on gastric cancer survival. AB - To study the effect of residual microscopic resection line disease in gastric cancer, we compared 47 patients with positive margins to 572 patients who underwent R0 resections using a multivariate analysis of factors affecting outcome. Although the presence of positive margins was a significant and independent predictor of outcome for the entire group (N = 619), this factor lost significance in patients who had undergone D2 or D3 lymph node dissections (N = 466). Subset analysis within the D2/D3 group determined that this finding was limited mainly to those patients with.5 positive nodes (N = 189). The survival of patients who had 5 positive nodes. We conclude that the significance of a positive microscopic margin in gastric cancer is dependent on the extent of disease. This factor is not predictive of outcome in patients who have undergone complete gross resection and have pathologically proved advanced nodal disease. Thus the goal in these cases should be an R0 resection when feasible but with the realization that the presence of >/=5 positive nodes (N2 disease according to the 1997 American Joint Committee on Cancer criteria) will mainly determine outcome and not microscopic residual cancer at the margin. PMID- 10457321 TI - Influence of p53 on herpes simplex virus type 1 vectors for cancer gene therapy. AB - Herpes simplex virus type 1 (HSV 1) vectors are under investigation for use in gene therapy for colorectal cancer liver metastases. Approximately 60% of colorectal cancers possess p53 mutations, and p53 mutations can cause tumor cell resistance to radiation therapy and chemotherapy. p53 is also known to co localize with at least one HSV 1 protein and influence HSV 1 gene expression. The purpose of this study was to determine if the loss or mutation of p53 in tumor cells alters the cytotoxicity of HSV 1 vectors. HSV 1 vector-mediated in vitro cytotoxicity assays were performed using stable transfectants of SAOS-2-LM2 cells and WiDr cells that express no p53, wild-type p53, mutant p53, or both wild-type p53 mutant p53. All stable transfectants were equally susceptible to HSV 1 vector cytotoxicity, and cell lines with mutant p53 were not resistant to HSV 1 vectors. These results provide additional rationale for the application of HSV 1 vector gene therapy for colorectal cancer. PMID- 10457322 TI - Role of alpha- and beta-calcitonin gene-related peptide in postoperative small bowel ileus. AB - Ablation of a-calcitonin gene-related polypeptide (CGRP) containing neurons with the afferent neurotoxin capsaicin improves postoperative foregut transit in a rodent model. Similarly, administration of a selective alpha-CGRP antibody or hCGRP((8-37)), a CGRP receptor antagonist, improves postoperative gastric emptying. Unlike the stomach, which contains only alpha-CGRP, the small bowel additionally contains beta-CGRP. The role of the latter in postoperative small bowel transit is unknown. The purpose of this study was to evaluate the effect of an alpha-CGRP antibody and hCGRP((8-37)) on postoperative small bowel transit. Male Sprague-Dawley rats underwent placement of duodenal catheters and were randomly assigned to 1 of 11 groups. Four groups were pretreated with 1% capsaicin. One week later, all animals underwent standardized laparotomy following administration of a control antibody or the alpha-CGRP mono-clonal antibody, or during infusion of hCGRP((8-37)) at varying doses. Small bowel transit was measured 25 minutes postoperatively. The alpha-CGRP antibody sped postoperative transit when given alone or in combination with capsaicin. In contrast, animals treated with hCGRP((8-37)) showed no significant improvement in postoperative transit, and the beneficial effect of capsaicin was blocked. Unlike their similar effects on postoperative gastric emptying, we found that hCGRP((8 37)) and the alpha-CGRP antibody had differing effects on postoperative small bowel transit. The reason for this is unknown but may be related to their differing specificities for alpha- and beta-CGRP. PMID- 10457323 TI - Laparoscopic cholecystectomy as a "true" outpatient procedure: initial experience in 130 consecutive patients. AB - Laparoscopic cholecystectomy has received nearly universal acceptance and is currently considered the "gold standard" for the treatment of cholelithiasis. Many centers have employed "short-stay" units or "23-hour admissions" for postoperative observation following laparoscopic cholecystectomy. The practice of early discharge as "true" outpatients following this procedure has not been well defined. A retrospective analysis of 130 consecutive patients undergoing laparoscopic cholecystectomy in an outpatient surgery unit was performed. A follow-up telephone survey was carried out of patients who successfully completed the procedure as outpatients. One hundred thirty patients underwent outpatient laparoscopic cholecystectomy. The patient population consisted of 78% women, with an age range of 17 to 76 years (mean age 47.1 years). Symptomatic gallstone disease was the indication for laparoscopic cholecystectomy in 92% of the patients. All patients underwent successful completion of laparoscopic cholecystectomy with no conversions to an open procedure. The mean length of operation was 75 +/- 23 minutes (range 25 to 147 minutes). The mean length of stay in the postanesthesia care unit (PACU) ranged from 95 to 460 minutes with a mean length of stay of 200 +/- 79 minutes. A total of eight patients (6.2%) were admitted to the hospital directly from the PACU in the immediate postoperative period. Six of these eight patients were discharged on the first postoperative day. Following discharge from the PACU, an additional six patients (4.6%) required hospital admission. Three of these six patients were discharged after a single day of hospitalization. Ninety-eight of 116 eligible patients were available for follow-up telephone evaluation. The outpatient experience was rated as good by 75.5% of the patients, fair by 22.5%, and poor by 2%. In retrospect, 20.4% of the patients stated that they would have preferred an inpatient to an outpatient procedure. Laparoscopic cholecystectomy can be performed as a true outpatient procedure with patients discharged to home within hours of completion of the procedure. Less than 10% of patients will fail this protocol and another 5% of the patients may require hospitalization after returning to their homes. PMID- 10457324 TI - Early laparoscopic cholecystectomy for acute cholecystitis: a safe procedure. AB - Acute cholecystitis is increasingly managed by laparoscopic cholecystectomy. Some reports have shown conversion and complication rates that are increased in comparison to elective laparoscopic cholecystectomy. This study reviews the combined experience of two hospitals where the intention was to perform early laparoscopic cholecystectomy for acute cholecystitis. A total of 152 cases of laparoscopic cholecystectomy for acute cholecystitis (evidence of acute inflammation clinically and pathologically) were identified. Conversion to open cholecystectomy was required in 14 cases (9%) in the total series. Laparoscopic cholecystectomy was performed within 2 days of admission in 76% (115 of 152) of patients. Conversion was significantly less likely in patients undergoing laparoscopic cholecystectomy within 2 days of admission (4 of 115) compared to those undergoing surgery beyond 2 days (10 of 37; P<0.0001). Eleven patients (7%) had postoperative complications; however, there were no cases of injury to the biliary system and no perioperative deaths. This series shows that laparoscopic cholecystectomy can be performed safely in patients with acute cholecystitis and suggests that early laparoscopic cholecystectomy is preferable to delaying surgery. Although the conversion rate to open surgery is higher than for elective cholecystectomy, the majority of patients (91%) still derive the well-recognized benefits of laparoscopic cholecystectomy. Early laparoscopic cholecystectomy is an acceptable approach to acute cholecystitis for the experienced laparoscopic surgeon. PMID- 10457325 TI - Selective decreases in levels of mRNA encoding a water channel (AQP3) in ileal mucosa after ileostomy in the rat. AB - Water channels (aquaporins) provide pathways for water permeation in a variety of epithelia. Aquaporin-3 (AQP3) has been localized to the basolateral membranes of epithelial cells in the small intestine, but mechanisms that regulate its expression and function have not been explored. To determine whether luminal content may influence intestinal AQP3 gene expression, adult Sprague-Dawley rats underwent sham laparotomy (N = 11) or loop ileostomy (N = 9) and were killed 8 days after procedures. Northern blot analysis was used to measure messenger RNA (mRNA) levels for AQP3 and the Na(+)/K(+) ATPase, a housekeeping transporter that regulates cellular levels of Na(+) and K(+). At sacrifice, histologic examination revealed only minimal changes in mucosal morphology. In sham animals, Na/K mRNA levels increased moderately in distal regions of the small intestine. Ileostomy did not alter these levels in any region. In contrast, in sham animals, AQP3 mRNA levels increased along the length of the intestine and were markedly higher in the distal ileum. Diversion of luminal contents decreased AQP3 mRNA levels in the postileostomy region by 30% to 50%. These findings indicate regional variations in expression of the AQP3 water channel in mucosa of the small intestine. In addition, they suggest that AQP3 gene expression may depend on the presence of luminal contents. PMID- 10457326 TI - Role of angiography and embolization for massive gastroduodenal hemorrhage. AB - The role of mesenteric angiography and embolization for massive gastroduodenal bleeding is unclear. We reviewed the records of patients who underwent angiography for acute, nonmalignant, and nonvariceal gastric or duodenal hemorrhage that was documented but not controlled by endoscopy. Fifty patients were identified over a 7-year period ending in March 1998. Only 17 patients (34%) were originally admitted to the hospital with gastrointestinal bleeding. All required treatment in the intensive care unit (mean 15 days) with a mean APACHE III score of 79 (29% predicted hospital mortality), and 32 (64%) had organ failure. A mean of 2.1 endoscopies were performed to locate the source of acute duodenal bleeding in 37 (74%) and gastric bleeding in 13 (26%). An average of 24.3 units of packed red blood cells were transfused per patient. Twenty-five patients (50%) were found to have active bleeding at angiography; all were treated by embolization as were 22 who underwent empiric embolization. Twenty-six patients (52%) were successfully treated by embolization and thus spared imminent surgery. Multiple variables were compared between those who were successfully treated by embolization and those considered failures. Time to angiography was considerably shorter (2.5 vs. 5.8 days, P<0. 017) and fewer total units of packed red blood cells were used (14.6 vs. 34, P<0.003) in those who were successfully treated. There was also a strong trend toward using fewer units of packed red blood cells for transfusion prior to angiography (11.2 vs. 17.1, P<0.08). No differences were found that could be attributed to gastric vs. duodenal sources, number of comorbid diseases, organ failure, APACHE score, age, or whether active bleeding was found at angiography. A total of 20 patients (40%) died including 9 of 17 patients operated on in an attempt to salvage angiographic failure. In summary, angiographic embolization should be performed early in the course of bleeding in otherwise critically ill patients. PMID- 10457327 TI - Appendectomy in the pre- and postlaparoscopic eras. AB - The role of laparoscopic appendectomy remains controversial since many authors have suggested that overall morbidity is primarily a function of the degree of appendicitis rather than the operative approach. We have reviewed our appendectomy experience to determine the advantages and/or disadvantages of the laparoscopic technique in cases of acute appendicitis, and furthermore to ascertain whether the extent of disease should affect the surgical approach used. Data were accumulated for all 1158 patients who underwent appendectomy at a single institution during the following three time periods that span the pre- and postlaparoscopic eras: period I (1987 to 1990), period II (1991 to 1993), and period III (1994 to 1997). Cases were categorized with regard to pathologic findings and operative approach (i.e., open or laparoscopic appendectomy). The percentage of appendectomies performed laparoscopically increased with time (0%, 27%, and 79% for periods I, II, and III, respectively). Overall, the total operating room time was slightly shorter for laparoscopic compared to open appendectomy (99 vs. 102 minutes; P<0.05). Operating room times for open appendectomy remained unchanged, but the times for laparoscopic appendectomy decreased from period II to period III (119 to 94 minutes; P<0.001). In cases of gangrenous/perforated appendicitis, the times for laparoscopic appendectomy were significantly shorter than those for open appendectomy (98/115 vs. 120/125 minutes; P<0.001 for both). Overall, the hospital stay was shorter for patients undergoing laparoscopic appendectomy (1.63 vs. 4.21 days; P<0.001), and the difference was maintained in all three time periods. The differences in length of hospital stay for lap-aroscopic vs. open appendectomy were most dramatic in gangrenous/perforated cases (1.8/3.0 vs. 4.0/9.0 days; P<0.001), whereas there was only a slight difference in cases of simple appendicitis, for example, 1.6 vs. 2.1 days (laparoscopic vs. open appendectomy, period III). There was a significant decrease in the percentage of perforated cases in which surgical treatment had been delayed (>8 hours) (21%, 5%, and 5%) over the three time periods, but the rate of "negative" appendectomies was similar (10%, 8%, and 8%). The complication rates following laparoscopic and open appendectomies during period II were 5.4% and 7.5%, respectively (P>0.05). Laparoscopic appendectomy results in a marked decrease in the length of hospital stay and similar postoperative morbidity compared to open appendectomy. In cases of gangrenous or perforated appendicitis, laparoscopic appendectomy appears to be especially worthwhile in regard to both operating room time and hospital stay. PMID- 10457328 TI - Bak expression and cell death occur in peritumorous tissue but not in pancreatic cancer cells. AB - Bak is a pro-apoptotic member of the Bcl-2 family whose genes are involved in regulation of programmed cell death. Using in situ hybridization, immunohistochemistry, and Northern blot analysis, we studied the expression of Bak in specimens from 12 normal pancreata and 26 primary pancreatic cancers, and correlated the findings with the clinical and histopathologic data of the patients. By comparison with normal pancreas, Northern blot analysis demonstrated a 2.5-fold increase of Bak messenger RNA expression in the tumor samples (P <0. 001). Elevated levels were found in 15 of the 26 pancreatic cancer tissue specimens. In these samples Bak expression was increased 4.3 fold (P <0.001). No association was detected between Bak expression and tumor stage. In situ hybridization and immunohistochemistry revealed that the tumor cells themselves and the stroma cells expressed only low levels of Bak. In contrast, in regions adjacent to the tumor, which showed chronic inflammation, there was always high expression in the acinar and inflammatory cells, explaining the increased Bak levels found in the tumor samples by means of Northern blot analysis. In the normal pancreas the expression of Bak was generally moderate in the acinar cells and low in the ductal and islet cells. In situ analysis using the terminal deoxynucleotidyl transferase method further showed that there was extensive cell death in the peritumorous areas with chronic inflammation. Taken together, these results suggest that in pancreatic cancer Bak expression and programmed cell death are present in cells that are localized in regions of chronic inflammation surrounding the pancreatic cancer cells but not in the tumor cells themselves, a situation that may facilitate tumor growth and spread. PMID- 10457329 TI - Loss of matrix-dependent cytoskeletal tyrosine kinase signals may regulate intestinal epithelial differentiation during mucosal healing. AB - Intestinal epithelial restitution and the migratory phenotype appear regulated by the extracellular matrix. Since integrin-associated adhesion to matrix triggers tyrosine kinase activity, we hypothesized that matrix-specific tyrosine kinase signals might modulate the intestinal epithelial migratory phenotype, particularly via focal adhesion kinase. Caco-2 cells were seeded at two densities on collagen I, laminin, fibronectin, and tissue culture plastic. Four days later the first cells were confluent, whereas the second cells were not contact inhibited and expressed migratory lamellipodia. Cells were fractionated into membrane/cytoskeletal and cytosolic fractions. Cytoskeletal tyrosine kinase activity in static cells was matrix dependent and, unlike cytoscolic tyrosine kinase, correlated with adhesion, highest on collagen and lowest on plastic. Migrating cells exhibited matrix-dependent increases in cystosolic tyrosine kinase activity. Cytosolic changes in tyrosine kinase activity in motile cells exceeded membrane/cytoskeletal changes. However, matrix-dependent variations in increase in cytosolic tyrosine kinase activity correlated inversely with changes in cytoskeletal tyrosine kinase activity, suggesting cytoskeletal tyrosine kinase translocation to the cytosol during motility. Indeed cytoskeletal focal adhesion kinase activity decreased during migration on collagen. Tyrosine kinase inhibition by genistein both inhibited migration and stimulated expression of brush-border enzymes downregulated during motility. Although enterocyte-matrix interactions alter both cytosolic and cytoskeletal tyrosine kinase activity, matrix-dependent cytoskeletal events are likely to regulate adhesion and differentiation in static cells. Loss of matrix-dependent cytoskeletal tyrosine kinase signals such as focal adhesion kinase during restitution may trigger a phenotypic switch to the "dedifferentiated" migrating intestinal epithelial phenotype. PMID- 10457330 TI - Paraesophageal herniation as a complication following laparoscopic antireflux surgery. AB - Paraesophageal herniation of the stomach is a rare complication following laparoscopic Nissen fundoplication. We retrospectively reviewed our experience with 720 patients undergoing laparoscopic Nissen fundoplications. Seven patients were found to have postoperative paraesophageal hernias requiring reoperation. The clinical presentation, diagnostic workup, operative treatment, and outcome were evaluated. There were no deaths or procedure-related complications. Clinical presentation was recurrent dysphagia in four, nonspecific abdominal symptoms in one, and acute abdomen in one. One additional patient was asymptomatic. Preoperatively the correct diagnosis was able to be confirmed in four of six patients by barium esophagogram. Four patients underwent successful laparoscopic repair. Two patients had a thoracotomy including one conversion from laparoscopy to thoracotomy. One patient had a lap-arotomy to reduce an intrathoracic gastric volvulus. At a mean follow-up of 2.5 months no patient had further complications. Paraesophageal herniation is a rare complication following laparoscopic Nissen fundoplication and a definitive diagnosis is often difficult to establish. Early dysphagia after surgery should alert the surgeon to this complication. Redo laparoscopic surgery is feasible but an open procedure may be necessary. PMID- 10457331 TI - Staging laparoscopy for pancreatic cancer should be used to select the best means of palliation and not only to maximize the resectability rate. AB - Staging laparoscopy, based on the assumption that endobiliary stenting is the best palliation, allegedly saves an "unnecessary" laparotomy for incurable pancreatic cancer. Our aim was to determine survival of patients with clinically resectable pancreatic cancer that is found to be unresectable intraoperatively and thereby infer appropriate utilization of staging laparoscopy. A retrospective analysis was undertaken of 148 patients with ductal adenocarcinoma (1985 to 1992) with a clinically resectable lesion based on current imaging techniques. All were considered candidates for resection but were deemed unresectable at operation because of metastases to the liver (group I; 29 patients), the peritoneum (group II; 22 patients), or distant lymph nodes (group III; 44 patients) or because of vascular invasion (group IV; 53 patients). Overall median survival was 9 months (range 1 to 53 months), but by group was as follows: group I, 6 months; group II, 7 months; group III, 11 months; and group IV, 11 months. Individual comparisons showed shorter survival for patients with distant nodal, liver, or peritoneal metastases than with nodal or vascular involvement (P<0.03). Staging laparoscopy should be performed to identify patients with liver or peritoneal metastases who have an expected survival of approximately 6 months, in whom short-term endoscopic palliation is satisfactory. Extended laparoscopy to identify lymph node or vascular involvement is contingent upon which palliation (operative vs. endoscopic) is considered most appropriate. Because we believe operative bypass provides better, more durable palliation in this latter group, we have not adopted extended laparoscopy. PMID- 10457332 TI - Pancreaticoduodenectomy for metastatic tumors to the periampullary region. AB - Although operative resection of metastatic lesions to the liver, lung, and brain has proved to be useful, only recently have there been a few reports of pancreaticoduodenectomies in selected cases of metastases to the periampullary region. In this report we present four cases of proven metastatic disease to the periampullary region in which the lesions were treated by pancreaticoduodenectomy. Metastatic tumors corresponded to a melanoma of unknown primary site, choriocarcinoma, high-grade liposarcoma of the leg, and a small cell cancer of the lung. All four patients survived the operation and had no major complications. Two patients died of recurrence of their tumors, 6 and 63 months, respectively, after operation; the other two patients are alive 21 and 12 months, respectively, after operation. It can be inferred from this small but documented experience, as well as a review of the literature, that pancreaticoduodenectomy for metastatic disease can be considered in selected patients, as long as this operation is performed by experienced surgeons who have achieved minimal or no morbidity and mortality with it. PMID- 10457333 TI - Fungal pseudotumor masquerading as pancreatic cancer-a sequela of new technology? AB - Fungal infection resulting in chronic pancreatitis is rare. We report a case of chronic pancreatitis due to fungal infection causing common bile duct obstruction and abdominal pain mimicking pancreatic cancer. Treatment included resection to cure the pain and rule out malignancy. Long-term effects of fungal infection may be seen more frequently as total parenteral nutrition, antibiotics, and foreign bodies (e.g., stents, drains, central venous catheters) are more often being used in the treatment of many diseases. PMID- 10457335 TI - Cholecystokinin-A receptor messenger RNA expression in human pancreatic cancer. AB - Cholecystokinin (CCK) is a gut peptide hormone known to stimulate postprandial gallbladder contraction and pancreatic enzyme secretion. It has also been shown to induce the growth of normal pancreas and of malignant and premalignant lesions in rodents. Although CCK has been shown to promote the growth of human adenocarcinoma cell lines, its role in the growth of human pancreatic adenocarcinomas in vivo is less clear. Localization of CCK receptors to neoplastic cells within resected human tissue specimens would be suggestive of its potential action as an in vivo promoter of human pancreatic cancer. Resected tissue specimens of pancreatic adenocarcinomas were therefore studied by both reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ hybridization for the presence of CCK-A receptors. Ninety percent of studied tumors demonstrated CCK-A expression by RT-PCR, and this expression was localized to neoplastic cells by in situ hybridization. An increase in the expression of CCK receptors is a mechanism by which pancreatic malignancies may gain a significant growth stimulus. PMID- 10457334 TI - G207, modified herpes simplex virus type 1, kills human pancreatic cancer cells in vitro. AB - Pancreatic cancer is often fatal, and further effective therapeutic options are needed. This study was designed to assess whether the replication-restricted herpes simplex virus, G207, was effective in killing human pancreatic cancer cells in vitro. G207, a multimutated strain of herpes simplex virus type 1 carrying lacZ reporter gene, is capable of efficient cytolytic growth in many dividing cells, including certain tumor cells, but not in nondividing cells. Three human pancreatic cell lines, AsPC-1, MIA PaCa-2, and BxPC-3, were infected with G207 at different multiplicities of infection. After 24 hours, expression of the lacZ reporter gene was tested using a histochemical X-gal assay. In addition, cell lines were infected with G207 for 24 to 48 hours; then the virus obtained from cell pellets and media supernatant was used to infect Vero cells to obtain G207 titers by plaque assay. To assess whether increasing viral immediate early gene expression would improve cytolysis and virus production, similar experiments were performed with the addition of 0.5 mmol/L of hexamethylene bisacetamide (HMBA) 1 hour after viral infection. Finally, MTS cell viability assays were performed to measure viable cells at 24 to 96 hours post infection. The X-gal assay data revealed a viral dose-dependent b-galactosidase expression, indicating G207 infectivity and expression of the lacZ reporter gene. Plaque assays demonstrated a viral dose-dependent increase in plaque formation, indicating viral production from all three cell lines. In addition, HMBA data indicated a modest increase in viral production. The MTS assay data indicated a dose dependent cytotoxicity for G207 in the cell lines tested. G207 infects, replicates in, and is cytotoxic to the above-listed human pancreatic cell lines in vitro and warrants therapeutic evaluation in models of pancreatic cancer. PMID- 10457336 TI - Discontinuous appendiceal involvement in ulcerative colitis: pathology and clinical correlation. AB - Continuous mucosal involvement from the rectum proximally is one of the hallmarks of ulcerative colitis. However, recent pathologic series report appendiceal ulcerative colitis in the presence of a histologically normal cecum, representing a "skip" lesion. The clinical significance of this finding has not been established. Eighty patients, 54 males and 26 females, average age 37.9 years (range 14 to 82 years) who underwent proctocolectomy for ulcerative colitis from January 1990 to September 1995 were examined to determine the rate of discontinuous appendiceal involvement. Excluded were 12 patients with prior appendectomy and 11 with fibrotic obliteration of the appendiceal lumen. Of the remaining 57 patients, seven (12.3%) had clear appendiceal involvement in the presence of a histologically normal cecum. These seven patients clinically were indistinguishable from the 50 patients without skip involvement of the appendix in terms of age at surgery, pretreatment medications, type of surgery, interval from diagnosis to definitive procedure, complications, functional results, and clinical course. Discontinuous appendiceal involvement was found in 12.3% of patients undergoing proctocolectomy for ulcerative colitis, and clinically these patients behave as those without this feature. PMID- 10457337 TI - Clinical subtypes of Crohn's disease according to surgical outcome. AB - Patients with Crohn's disease are typically classified into perforator or nonperforator groups. The perforator group includes those who present with acute perforation, fistulas, or abscess formation. The nonperforator group presents with stricture, obstruction, or unresponsiveness to medical therapy. Our purpose was to investigate whether perianal disease constitutes a separate predictor of surgical outcome. The form of presentation was classified as perforator, nonperforator, or perianal disease in 91 patients undergoing 232 operations for Crohn's disease. Those with perforating complications presented with the highest Crohn's Disease Activity Index, followed by those with nonperforating complications, and then the perianal disease group. However, the perianal disease group appeared to have the most rapid rate of recurrence and subsequent surgery, followed next by the perforator, and then the nonperforator group. Recurrence rate and subsequent operation intervals for the perforator group appeared to lengthen when those patients were treated with steroids and/or immunosuppressants, as compared to nonsteroidal and/or antimicrobial agents. Recurrence rate and subsequent operation intervals appeared to lengthen for the nonperforator and perianal disease groups when they were treated with nonsteroidal and/or antimicrobial therapy, as compared to steroids and/or immunosuppressants. Our data indicate that perianal disease, as a form of presentation of Crohn's disease, has independent predictive value, although this is not accurately reflected by the Crohn's Disease Activity Index. PMID- 10457338 TI - Surgical feeding gastrostomy: are we overdoing it? AB - Feeding gastrostomy is a commonly performed procedure in North America. Our aim was to study the outcome of patients undergoing feeding gastrostomy to better define patients who will benefit from the procedure as opposed to those in whom it may be futile. A cohort of the most recent 100 consecutive patients undergoing feeding gastrostomy in a community teaching hospital was retrospectively studied. The main indication for gastrostomy was neurologic disorder interfering with eating/swallowing (group A-54 patients), followed by debilitating systemic disease (group B-26 patients) and obstructive malignancy of the head and neck or esophagus (group C-20 patients). Forty-one patients died within 30 days of the procedure (41%). The overall 30-day survival rates in groups A, B, and C were 70%, 15%, and 85%, respectively. In four patients death was caused by intraperitoneal leak from the gastrostomy site; the remaining patients died of their underlying disease. Five patients required reoperation for gastric leakage around the gastrostomy within 30 days. Only nine patients could be traced who were alive and still using the gastrostomy a year after its placement: two in group A, none in group B, and seven in group C. APACHE II scores at tube insertion also predicted survival; 30-day survival rates in patients with scores of 10 and below, 11 to 15, 16 to 20, and over 20 were 96%, 71%, 48%, and 18%, respectively. No patient with an APACHE score above 15 belonging to group B (debilitating disease) survived more than 30 days. We conclude that to have a beneficial therapeutic effect feeding gastrostomy should be performed selectively. Severe debilitating systemic conditions that interfere with normal eating, when combined with a high APACHE II score, indicate the futility of gastrostomy. PMID- 10457339 TI - Venting intraluminal drains in pancreaticoduodenectomy. AB - The utility of placing biliary, pancreatic, or enteric "venting"tubes (externally draining devices traversing the bowel or bile duct that have their distal tip located intraluminally near the biliary or pancreatic anastomosis) when performing a pancreaticoduodenectomy has received little attention to date. We hypothesize that these venting tubes do not decrease the morbidity or mortality associated with pancreaticoduodenectomy and may actually be a source of additional morbidity. To characterize our use of and the effect of these drains, we retrospectively analyzed 136 pancreaticoduodenectomies (127 partial, 9 total) performed over a 24-month period. Venting drain use, drain type and size, drain location, duration of intubation, hospital course, and postoperative complications were noted. Venting tubes were used in 80 patients (59%). The use of these drains had no significant relationship to postoperative length of stay, the development of major complications, overall morbidity, or mortality (P>0.05). Such drains also did not significantly shorten the length of hospital stay (P>0.05) or improve outcome when available to augment local control following luminal leak (n = 6) or regional abscess (n = 7). These drains were removed at a median interval of 29 days postoperatively (range 6 to 77 days). Seven patients had complications that were directly related to the venting drain; four of these patients had a documented intra-abdominal luminal leak from the site of drain removal, whereas the other three were hospitalized for presumed leakage secondary to immediate, severe abdominal pain following removal of the drain. These seven patients were elderly (mean age 70 years) and often harbored pancreatic ductal carcinoma (n = 6). Intraluminal drains afford no distinct advantage in terms of shortening the postoperative length of stay, decreasing operative morbidity and mortality, or improving local control with regional sepsis in pancreaticoduodenectomies. Furthermore, they may add an additional source of morbidity and we no longer employ them routinely. PMID- 10457340 TI - Characterization and reduction of ischemia/reperfusion injury after experimental pancreas transplantation. AB - Reperfusion injury after pancreas transplantation is a cause of early graft pancreatitis. The aim of this study was to quantify pancreatic microcirculation after pancreas transplantation in correlation with cold ischemia time. In a second step the effect of N-acetylcysteine on reperfusion damage was tested. Pancreas transplantation was performed in three different groups of male Lewis rats. Groups 1 and 2 received no special treatment. Cold ischemia time was 1.5 hours in group 1 and 16 hours in groups 2 and 3. In group 3 donor and recipient were both treated with N-acetylcysteine (300 mg/kg) 1.5 hours after reperfusion graft microcirculation was quantified by means of intravital microscopy. Rhodamine-labeled leukocytes, fluoroscein isothiocyanate-labeled erythrocytes, and fluoroscein isothiocyanate-albumin were used as fluorochromes. After a cold ischemia time of 16 hours, functional capillary density, erythrocyte velocity, and leukocyte-endothelium interaction were reduced significantly compared to a cold ischemia time of 1.5 hours (P<0.05). After 16 hours of cold ischemia, treatment with N-acetylcysteine improved all of these parameters (PG(1) transition; induced expression had immediate-early response characteristics and abruptly declined prior to the onset of DNA synthesis. Transcriptional activity of the PAI-1 gene paralleled the steady-state mRNA abundance profile during this first synchronized growth cycle after release from quiescence. Although PAI-1 mRNA levels were up-regulated (approximately threefold) upon exposure to several different growth factors, neutralizing antibodies to transforming growth factor-beta1 (TGF-beta1) effectively attenuated the more than ninefold serum-associated PAI-1 inductive response by more than 70% (at both the mRNA transcript and protein levels). Similar to the metabolic requirements for serum-mediated PAI-1 transcription, PAI-1 induction upon addition of TGF-beta1 to quiescent NRK cell cultures was actinomycin D sensitive and resistant to cyclohexamide and puromycin, suggesting a primary mode of transcript control. The response to protein synthesis inhibitors, however, was complex. While cyclohexamide appeared to stabilize, or at least maintain, fetal bovine serum (FBS)- or TGF-beta1-stimulated PAI-1 mRNA levels, puromycin had no such affect. The amplitude and duration of induced PAI-1 expression were the same in either the presence or absence of puromycin. Cyclohexamide when used alone (i.e., in non-FBS- or TGF-beta1-treated cultures), moreover, effectively stimulated PAI-1 induction whereas puromycin was ineffective. Although TGF-beta1 was not a complete mitogen in the NRK cell system, incubation of quiescent renal cell cultures with TGF-beta1, prior to serum stimulation, resulted in a 10- to 12 fold increase in PAI-1 expression coincident with exit out of G(0). These data support a model in which PAI-1 gene expression is closely associated with creation of the growth-activated state and that cell cycle controls appear to be superimposed on the time course of the serum-induced expression of the PAI-1 gene. PMID- 10457358 TI - Rate constants of acetylcholine receptor internalization and degradation in mouse muscles. AB - The rate constants for internalization and subsequent extrusion of acetylcholine receptors (AChRs) during degradation in adult innervated and denervated mouse diaphragm muscles were determined using proteinase K (PK) digestion. This procedure separated (125)I-alpha-bungarotoxin (Bgt)-labeled AChRs into PK sensitive and PK-resistant compartments. The time course of the residual radioactivity in these two compartments suggested that they represented surface membrane and internalized compartments, respectively. The data were compatible with a mathematical model based on the assumption that during degradation of AChRs a surface compartment, A, fed an internal compartment, B, with an internalization rate constant (k(i)), and that B is drained from the cell with an extrusion rate constant (k(o)). Using the mathematical model, we were able to determine that k(i) and k(o) were, respectively, 0.068 (t(1/2) approximately 10.2 days) and 0.69-0.55 (t(1/2) approximately 1.0- 1.25 days) for innervated muscle and were, respectively, 0.69 (t(1/2) approximately 1.0 day) and 6.93 (t(1/2) approximately 0.1 day) for denervated muscle. Thus, the rate for internalization was about 8-10 times slower than that for extrusion from the cell for both the slowly degrading innervated (Rs) AChRs and for the rapidly degrading denervated (Rr) AChRs. This inequality between k(i) and k(o) therefore allows the combined quantity of A(t) + B(t), usually measured in AChR degradation studies, to approximate a single exponential. PMID- 10457359 TI - Protein kinase C activator PMA reduces the Ca(2+) response to antigen stimulation of adherent RBL-2H3 mucosal mast cells by inhibiting depletion of intracellular Ca(2+) stores. AB - Activation of protein kinase C has been shown to reduce the Ca(2+) responses of a variety of cell types. In most cases, the reduction is due to inhibition of Ca(2+) influx, but acceleration of Ca(2+) efflux and inhibition of Ca(2+) store depletion by protein kinase C activation have also been described. For adherent RBL-2H3 mucosal mast cells, results from whole-cell patch clamp experiments suggest that protein kinase C activation reduces Ca(2+) influx, while experiments with intact, fura-2-loaded cells suggest that Ca(2+) influx is not affected. Here we present single-cell data from Ca(2+) imaging experiments with adherent RBL-2H3 cells, showing that antigen-stimulated Ca(2+) responses of phorbol 12-myristate 13-acetate (PMA)-treated cells are more transient than those of control cells. PMA also reduced the response to antigen in the absence of extracellular Ca(2+), indicating that depletion of intracellular Ca(2+) stores is inhibited. If PMA was added after stores had been depleted by thapsigargin, a small decrease in [Ca(2+)](i) was observed, consistent with a slight inhibition of Ca(2+) influx. However, the major effect of PMA on the antigen-stimulated Ca(2+) response is to inhibit depletion of intracellular Ca(2+) stores. We also show that inhibition of protein kinase C did not enhance the Ca(2+) response to antigen, suggesting that inhibition of the Ca(2+) response by activation of protein kinase C does not contribute to the physiological response to antigen. PMID- 10457361 TI - Alterations in the glycolytic and glutaminolytic pathways after malignant transformation of rat liver oval cells. AB - Oval cells are liver epithelial cells that proliferate during the early stages of hepatocarcinogenesis induced by a variety of chemicals. The oval cell lines OC/CDE 6 and OC/CDE 22 have been established in our laboratory at two time points (6 and 22 weeks) of the carcinogenic process and have been malignantly transformed by different procedures. During the transformation process, the glycolytic and glutaminolytic flux rates were consistently up-regulated and this process was accompanied by an overproportional increase in the activities of cytosolic hexokinase and 6-phosphogluconate dehydrogenase. In transformed oval cells, a strong correlation between the glycolytic flux rate and glutamine consumption as well as glutamate production was observed. Furthermore, the transport of glycolytic hydrogen, produced by the glyceraldehyde 3-phosphate dehydrogenase-catalyzed reaction, from the cytosol into the mitochondria by means of the malate-aspartate shuttle was enhanced, this being due to alterations in the activities of malate dehydrogenase and glutamate oxaloacetate transaminase. The up-regulation of the glycolytic hydrogen transport and the alterations in the glycolytic enzyme complex led to an enhanced pyruvate production at high glycolytic flux rates. Taken together, our data are further proof that a special metabolic feature (increased glycolysis and glutaminolysis) is characteristic for tumor cells and that the mechanisms by which this metabolic state is induced can be totally different. PMID- 10457360 TI - Synthetic peptide sequence from the C-terminus of the insulin-like growth factor I receptor that induces apoptosis and inhibition of tumor growth. AB - Although the type 1 insulin-like growth factor receptor (IGF-IR) is a potent inhibitor of apoptosis, its C-terminus sequence sends contradictory signals, including a clearly proapoptotic signal. We have synthesized a peptide, peptide 2, having the sequence of the IGF-IR from residue 1282 to residue 1298 (C terminus of the beta subunit). To favor its uptake into cells, we linked it to a stearic acid moiety at its NH-terminus. Peptide 2 is taken up by the cells, where it inhibits DNA synthesis and causes apoptosis, while a scrambled peptide (with stearic acid) and peptide 2 without stearic acid are completely ineffective. Peptide 2 is more effective when cells are in anchorage-independent conditions than when they grow in monolayer cultures. Accordingly, we find that peptide 2 can inhibit the growth of a human prostatic cell line in nude mice. The proapoptotic effect of peptide 2 is inhibited by the expression of Bcl-2 or by a dominant negative mutant of caspase 9. These and other data indicate that peptide 2 does not seem to be competing directly with the IGF-IR for common substrates, but that its proapoptotic effect is related to its ability to activate the caspase cascade. PMID- 10457362 TI - 13C-NMR investigation of protein synthesis during apoptosis in human leukemic cell lines. AB - In order to evaluate the role of protein synthesis in apoptosis, (13)C-NMR has been used to study the levels of protein synthesis in three different human leukemic cell lines in the presence and absence of dexamethasone-induced apoptosis. Measurements were done on one dexamethasone-sensitive (CEM-C7-14) and two different dexamethasone-resistant variants (CEM-4R4 and CEM-ICR27-4). The incorporation of (13)C-labeled amino acids into cellular proteins, which reflects the level of new protein synthesis, was monitored by (13)C-NMR spectroscopy. In the absence of dexamethasone, the level of protein synthesis was found to be significantly different among the three cell lines. Dexamethasone caused a significant reduction ( congruent with 60-87%) in the level of protein synthesis in dexamethasone-sensitive CEM-C7-14 cells, while having no significant effect on protein synthesis in dexamethasone-resistant CEM-4R4 cells. Dexamethasone treatment caused a significant enhancement of the level of protein synthesis in the CEM-ICR27-4 cells. Synthesis of proteins was found to occur during apoptosis, albeit at a low level, suggesting a role for the synthesis of specific proteins in the mechanism of apoptosis. PMID- 10457363 TI - Role and interaction of connective tissue growth factor with transforming growth factor-beta in persistent fibrosis: A mouse fibrosis model. AB - Skin fibrotic disorders are understood to develop under the influence of some growth factors, such as transforming growth factor-beta (TGF-beta), basic fibroblast growth factor (bFGF), or connective tissue growth factor (CTGF). To establish an appropriate animal model of skin fibrosis by exogenous application of growth factors, we investigated the in vivo effects of growth factors by injecting TGF-beta, CTGF, and bFGF into the subcutaneous tissue of newborn mice. A single application of TGF-beta or bFGF resulted in the formation of transient granulated tissue that disappeared despite 7 days of consecutive injections. A single CTGF injection also caused slight granulation. However, injecting TGF-beta plus CTGF produced long-term fibrotic tissue, which persisted for at least 14 days. Also, fibrotic tissue was observed when CTGF was injected from 4 to 7 days after TGF-beta injections for the first 1-3 days. In situ hybridization analysis revealed the expression of CTGF mRNA in the fibroblasts at least in a few fibrotic conditions. These findings suggest that either CTGF mRNA or an application of exogenous CTGF protein is required for the development of persistent fibrosis. From our study, it appears that interaction of several growth factors is required for persistent fibrotic tissue formation, with TGF beta causing the induction and CTGF needed for maintenance of skin fibrosis. The animal model on skin fibrosis by exogenous application of growth factors developed in this study may prove useful for future studies on fibrotic disorders. PMID- 10457364 TI - Regulation of myelin basic protein gene transcription by Sp1 and Puralpha: evidence for association of Sp1 and Puralpha in brain. AB - Direct interaction between transcription factors may provide a mechanism for the regulatory function of these proteins on transcription of the responsive genes. These interactions may be facilitated if the target DNA sequences for the participant regulatory proteins are overlapped or positioned in close proximity to each other within the promoter of the responsive genes. In earlier studies, we identified a cellular protein, named Puralpha, which upon binding to the MB1 regulatory DNA sequence of the myelin basic protein (MBP) gene, stimulates its transcription in central nervous system (CNS) cells. Here, we provide evidence for binding of the ubiquitous DNA binding transcription factor, Sp1, to the MB1 DNA motif at the region that partially overlaps with the Puralpha binding site. We demonstrate that binding of Puralpha to its target sequence is enhanced by inclusion of Sp1 in the binding reaction. Under this condition, binding of Sp1 to the MB1 regulatory sequence remained fairly unchanged, and no evidence for the formation of Puralpha:MB1:Sp1 was observed. This observation suggests that transient interaction of Puralpha and Sp1 may result in stable association of Puralpha and the MB1 element. In support of this notion, results from immunoprecipitation/Western blot studies have established association of Puralpha and Sp1 in nuclear extracts from mouse brain. Of interest, Puralpha appears to bind to the phosphorylated form of Sp1 which is developmentally regulated and that coincides with the periods when MBP gene expression is at its maximum level. Results from cotransfection studies revealed that ectopic expression of Puralpha and Sp1 synergistically stimulates MBP promoter activity in CNS cells. The importance of these findings in stage-specific expression of MBP during brain development is discussed. PMID- 10457365 TI - Positive and negative regulation of chondrogenesis by splice variants of PEBP2alphaA/CBFalpha1 in clonal mouse EC cells, ATDC5. AB - The alphaA type of the alpha subunit of the polyomavirus enhancer binding protein 2 (PEBP2alphaA), also called the core binding factor alpha1 (CBFalpha1) or til-1, plays crucial roles in osteogenesis. Little is known, however, about the function of PEBP2alphaA in chondrogenesis. Here, we examined the role of PEBP2alphaA in chondrogenesis of clonal mouse embryonal carcinoma cells, ATDC5, which are committed as chondroprogenitors. We found that as ATDC5 cells condensed and formed cartilaginous nodules, PEBP2alphaA increased, and the level was maintained throughout the process of chondrocytic maturation. When an established dominant negative form of PEBP2alphaA was introduced in undifferentiated ATDC5 cells, the cellular condensation and the subsequent processes were inhibited. This inhibition was overcome with BMP-4 treatment, which increased the endogenous expression of PEBP2alphaA. Thus, the process of chondrogenesis is regulated by the level of PEBP2alphaA activity. Along with the wild-type PEBP2alphaA, a splice variant form, til-1 G2, is naturally expressed in ATDC5 cells. In luciferase reporter assays, til-1 G2 not only exhibited a limited ability to transactivate the osteocalcin promoter but also inhibited the activity achieved by the wild type PEBP2alphaA. When til-1 G2 was overexpressed by stable transfection in undifferentiated ATDC5 cells, it inhibited the progression of chondrogenesis. Therefore, we conclude that PEBP2alphaA acts as a positive regulator of chondrogenesis, and that this positive effect may be finely tuned by the opposing effect of the til-1 G2 isoform. PMID- 10457366 TI - Regenerative potential of adult O1+ oligodendrocytes. AB - Remyelination in the adult central nervous system (CNS) is preceded by the generation of new oligodendrocytes (ODCs) but the source of the new ODCs has not been resolved. Adult galactocerebroside positive (O1+)ODCs proliferate when cultured with purified sensory neurons (Wood and Bunge, Nature 320:756-758, 1999), implying that differentiated ODCs could be an important source of new myelinating ODCs. To test this possibility purified O1+ODCs (>96% purity) were plated at low density (20-50 cells/culture) into cultures of purified dorsal root ganglion neurons. Three days after plating, single O1+ODCs were located (209 ODCs/43 cultures) and sequentially observed for 4 weeks. The ODCs began to proliferate by the fifth day after plating and formed large colonies by the third week. Most cells in these colonies were 01- but positive for another ODC antigen, O4. A few O1+, myelin basic protein (MBP)+ODCs, and glial fibrillary acidic protein (GFAP)+cells with astrocytic morphology were observed in some colonies. In similar cultures plated with cell-sorted O1+ODCs (>99.5% purity), ciliary neurotrophic factor (CNTF, 1ng/ml) increased the number and size of colonies, the number of O1+MBP+ODCs (including ODCs producing myelin-like profiles in association with axons) and the number of GFAP+ astrocytes, relative to untreated controls. The results are evidence that CNTF exerts a trophic effect on adult O1+ODCs, and/or their progeny, and that cells generated by division of O1+ODCs can become either new myelin-producing ODC, or astrocytes. This plasticity in regenerative potential of adult O1+ODCs has not been previously demonstrated. PMID- 10457367 TI - Subtle roles of neural cell adhesion molecule and myelin-associated glycoprotein during Schwann cell spiralling in P0-deficient mice. AB - Peripheral nerves of P0-deficient mice display a severe dysmyelinating phenotype, confirming the view that P0 mediates myelin formation and compaction. In addition to the compromised myelin organization, an elevated expression of several cell recognition molecules was described in the axon-Schwann cell units of P0 deficient mice. The present study was performed to focus on the subcellular localization and functional roles of two of these up-regulated molecules, the neural cell adhesion molecule (N-CAM) and the myelin-associated glycoprotein (MAG). We show by postembedding immunoelectron microscopy that in peripheral nerves of P0-deficient mice both molecules are expressed in noncompacted myelin like regions. In addition, N-CAM, but not MAG, is detected in partially compacted myelin. By the generation of P0/N-CAM- and P0/MAG-deficient double mutants, we investigated the roles of the dysregulated molecules in P0-deficient mice. In 4 week-old double mutants, the dysmyelinating phenotype of the axon-Schwann cell units was very similar to that seen in the P0-deficient single mutants, suggesting that neither N-CAM nor MAG are responsible for the poor myelin compaction in P0-deficient mice. However, the noncompacted turns surrounding the abnormally compacted regions were significantly reduced in number in P0/MAG mutants as compared to P0 or N-CAM/P0 mice. During formation of myelin sheaths, absence of N-CAM resulted in a transient retardation of Schwann cell spiralling in P0-deficient mice, whereas absence of MAG impaired Schwann cell spiralling for a more extended time period. Our findings demonstrate for the first time that MAG and also N-CAM can play significant roles during myelin formation in the peripheral nervous system. Because these functional roles are detectable only in the absence of P0, our results confirm the view that myelin-related molecules can play distinct, but also partially overlapping roles. PMID- 10457368 TI - K(+) inward rectifier currents in reactive astrocytes from adult rat brain. AB - We characterized inward rectifier (K(IR)) currents in reactive astrocytes activated by CNS injury. We used primary cultures of reactive astrocytes obtained from gelatin sponge implants in adult rat brains, a system that yielded highly purified, homogeneous cultures with >95% of cells positive for GFAP, vimentin, and S-100beta. Ionic channels were studied in 1-21-day-old primary cultures using a nystatin-perforated patch clamp technique. Fast Na(+) currents were identified in <2% of cells. Most cells exhibited outward currents positive to -50 mV, with one component being sensitive to charybdotoxin, iberiotoxin, and tetraethylammonium chloride, and another component being sensitive to 4 aminopyridine. Two populations of cells were distinguished, based on presence or absence of Ba(2+)-sensitive K(IR) current negative to the K(+) reversal potential (E(k)), with >80% of cells expressing K(IR) currents. In contrast to previous reports on mammalian astrocytes, the current-voltage curve showed no appreciable current between E(k) and -50 mV, reflecting strong rectification by K(IR) channels. The magnitude of K(IR) current at -130 mV (I(-)(130)) did not change significantly during 21 days in culture (123 cells), suggesting constitutive expression of K(IR) channels. The fraction of K(IR)-negative cells was not affected by serum-starvation for 16-24 h. In cells with I(-)(130) >/= -30 pA, the membrane potential was invariably near E(k) and depolarized appreciably on addition of Ba(2+), but in cells with I(-)(130) < -30 pA, resting potentials ranged from -40 mV to -90 mV. We conclude that most adult reactive astrocytes constitutively express K(IR) channel(s) that exhibit strong rectification not previously observed in mammalian astrocytes. PMID- 10457369 TI - Circumferential migration of ameboid microglia in the margin of the developing quail retina. AB - Central-to-peripheral migration of QH1-positive microglial precursors occurs in the vitrealmost part of the developing quail retina. This study shows that some QH1-positive ameboid cells with morphological features of migrating cells are already present in the margin of the retina before microglial precursors migrating centrally to peripherally arrive in this zone. Because the earlier cells are oriented parallel to the ora serrata, we deduce that some microglial cells migrate circumferentially in the margin of the retina, whereas other microglial precursors migrate from central to peripheral zones. Microglial cells that migrate circumferentially are first seen on embryonic day 6 (E6) and advance in a temporal-to-dorsal-to-nasal direction from the temporoventral quadrant of the retina. When cells migrating centrally to peripherally reach the retinal margin, they meet those migrating circumferentially. From E6 on, some QH1 positive dendritic cells in the ciliary body bear processes that penetrate the retina, where they are oriented circumferentially. These observations suggest that microglial cells that migrate circumferentially in the retinal margin share a common origin with dendritic cells of the ciliary body. Therefore, microglial cells of the quail retina appear to make up a heterogeneous population, with some cells originating from the pecten/optic nerve head area and others from the ciliary body. PMID- 10457370 TI - Rapid uptake and degradation of glycine by astroglial cells in culture: synthesis and release of serine and lactate. AB - Free glycine is known to have vital functions in the mammalian brain, where it serves mainly as both neurotransmitter and neuromodulator. Despite its importance, little is known about the metabolic pathways of glycine synthesis and degradation in the central nervous system. In this study, the pathway of glycine metabolism in astroglia-rich primary cultures from rat brain was examined. The cells were allowed to degrade glycine in the presence of [U-(14)C]glycine, [U (13)C]glycine or [(15)N]glycine. The resulting intra- and extracellular metabolites were analyzed both by high-performance liquid chromatography and by (13)C/(15)N nuclear magnetic resonance spectroscopy. Glycine was rapidly consumed in a process obeying first-order kinetics. The initial glycine consumption rate was 0.47 nmol per mg protein. The half-life of glycine radiolabel in the incubation medium was shorter than that of glycine mass. This suggests that glycine is produced from endogenous sources and released simultaneously with glycine uptake and metabolism. As the main metabolites of the glycine carbon skeleton in astroglia-rich primary cultures from rat brain, serine and lactate were released during glycine consumption. The main metabolite containing the glycine amino nitrogen was glutamine. To establish a metabolic pathway from glycine to serine in neural tissue, homogenates of rat brain and of neural primary cultures were assayed for their content of serine hydroxymethyltransferase (SHMT) and glycine cleavage system (GCS). SHMT activity was present in homogenates of rat brain as well as of astroglia-rich and neuron rich primary cultures, whereas GCS activity was detectable only in homogenates of rat brain and astroglia-rich primary culture. Of the two known SHMT isoenzymes, only the mitochondrial form was found in rat brain homogenate. It is proposed that, in neural tissue, glycine is metabolized by the combined action of SHMT and the GCS. Owing to the absence of the GCS from neurons, astrocytes appear to be the only site of this part of glycine metabolism in brain. However, neurons are able to utilize as energy source the lactate formed by astroglial cells in this metabolic pathway. PMID- 10457371 TI - Metastasis-associated mts1 (S100A4) protein is selectively expressed in white matter astrocytes and is up-regulated after peripheral nerve or dorsal root injury. AB - The S100 family of calcium binding proteins has been shown to be involved in a variety of physiological functions, such as regulation of enzyme function, cell motility, modification of extracellular matrix, and cell proliferation. Several members of the S100 family are expressed in the nervous system, but their functional roles are still largely obscure. The Mts1 gene codes for the S100A4 protein, which has been implicated in the control of cell proliferation and metastasis activity of tumor cells. We have used immunohistochemistry to examine the expression pattern of the Mts1 protein in the adult rat spinal cord and how this expression is influenced by peripheral nerve or dorsal root injury. Mts1 immunoreactivity (IR) was present only in white matter astrocytes in the intact spinal cord. Sciatic nerve as well as dorsal root injury induced a marked and prolonged up-regulation of Mts1-IR in astrocytes in the region of the dorsal funiculus containing the central processes of the injured primary sensory neurons. These findings suggest that Mts1 plays a unique physiological role in white matter astrocytes as well as in the response of astrocytes to degeneration of myelinated axons. PMID- 10457373 TI - Myelin formation by Schwann cells in the absence of beta4 integrin. AB - The interaction of the Schwann cell with its basal lamina has been hypothesized to be an important prerequisite for the formation of a myelin sheath in the peripheral nervous system. One possible player in this interaction is beta4 integrin; it is up-regulated during myelin formation and, in association with alpha6 integrin, can interact with particular components of the Schwann cell basal lamina. In order to characterize the functional roles of beta4 integrin during myelination, we investigated myelination in the absence of beta4 integrin, i.e., in peripheral nerve tissue from beta4 integrin-deficient mice. Because the mutants die within several hours after birth, we cultured dorsal root ganglia from neonatal mutants under conditions that promote myelination, quantified the myelin segments by immunofluorescence, and investigated the ultrastructure of the cultured myelin sheaths. In another approach, we quantified the few myelin sheaths that are detectable in femoral nerves of newborn animals. Based on both approaches, we conclude that myelination by Schwann cells can occur in the absence of beta4 integrin demonstrating that this Schwann cell component is dispensable for myelin formation in peripheral nerves. PMID- 10457372 TI - Expression of a homologue of rat NG2 on human microglia. AB - The expression of NG2 chondroitin sulfate has been widely associated with oligodendrocyte precursors in rodents. We used a monoclonal antibody (9.2.27) against the human homologue of the rat NG2 to determine whether expression of this molecule was associated with a specific glial cell population present in dissociated cell preparations derived from adult and fetal human brain tissue. Our data, derived using FACS and immunocytochemical analyses of immediately ex vivo or cultured glial cells, indicate that the large majority of NG2 expressing cells belonged to the microglial lineage (CD68, CD11c) rather than to the oligodendrocyte lineage (O4, A2B5, GalC). In situ immunohistochemistry performed on non-fixed normal spinal cord tissue confirmed the observation that NG2 is expressed by mononuclear phagocytes of the CNS. In contrast, peripheral blood derived monocytes were NG2(-). Cells from fetal brain tissue showed only small numbers of NG2(+) cells, which was consistent with the number of microglial cells in this preparation. In absence of additional markers, we cannot exclude that this anti-NG2 mAb might also recognize human oligodendrocyte progenitor cells. PMID- 10457374 TI - Activation of A(1) adenosine or mGlu3 metabotropic glutamate receptors enhances the release of nerve growth factor and S-100beta protein from cultured astrocytes. AB - Pharmacological activation of A(1) adenosine receptor with 2-chloro-N6 cyclopentyladenosine (CCPA) or mGlu3 metabotropic glutamate receptors with (2S,2'R,3'R)-2-(2', 3'-dicarboxycyclopropyl)glycine (DCG-IV) or aminopyrrolidine 2R, 4R-dicarboxylate (2R,4R-APDC) enhanced the release of nerve growth factor (NGF) or S-100beta protein from rat cultured astrocytes. Stimulation of release by CCPA and DCG-IV or 2R,4R-APDC was inhibited by the A(1) adenosine receptor antagonist 8-cyclopentyl-1, 3-dipropylxanthine and by the mGlu2/3 receptor antagonist (2S,1'S, 2'S,3'R)-2-(2'-carboxy-3'-phenylcyclopropyl)glycine (PCCG-4), respectively. Time-course studies revealed a profound difference between the release of S-100beta protein and the release of NGF in response to extracellular signals. Stimulation of S-100beta protein exhibited rapid kinetics, peaking after 1 h of drug treatment, whereas the enhancement of NGF release was much slower, requiring at least 6 h of A(1) adenosine or mGlu3 receptor activation. In addition, stimulation of NGF but not S-100beta release was substantially reduced in cultures treated with the protein synthesis inhibitor cycloheximide. In addition, a 6-8 h treatment of cultured astrocytes with A(1) or mGlu3 receptor agonists increased the levels of both NGF mRNA and NGF-like immunoreactive proteins, including NGF prohormone. We conclude that activation of A(1) adenosine or mGlu3 receptors produces pleiotropic effects in astrocytes, stimulating the synthesis and/or the release of protein factors. Astrocytes may therefore become targets for drugs that stimulate the local production of neurotrophic factors in the CNS, and this may provide the basis for a novel therapeutic strategy in chronic neurodegenerative disorders. PMID- 10457385 TI - Herpes simplex virus encephalitis: chronic progressive cerebral MRI changes despite good clinical recovery and low viral load - an experimental mouse study. AB - Cranial magnetic resonance imaging (MRI) is a sensitive diagnostic tool for the in vivo detection of morphological abnormalities in herpes simplex virus encephalitis (HSVE). We performed a long-term MRI study in a mouse model of HSVE. Cranial MRI findings were compared with the viral load within brain tissue, the presence of HSV DNA in the cerebrospinal fluid (CSF), a daily clinical assessment and post-mortem neurohistopathological studies. A 1.5 T cranial MRI scanner with standard spin-echo sequences was used. Viral load within the brain and the presence of HSV DNA in cerebrospinal fluid were determined by a polymerase chain reaction assay. Clinically, animals were severely affected within the first 2 weeks and recovered thereafter. Focal histopathological and MRI abnormalities involved predominantly limbic structures, a pattern that mimics human disease. Severity and extent of abnormalities had increased at 6 months despite clinical improvement. HSV DNA was present in CSF during the acute disease only. Brain viral load peaked at day 10 and declined thereafter. MRI as an in vivo monitoring approach may reveal chronic progressive changes in HSVE, despite clinical recovery and low viral load in the brain. Secondary, not directly virus-mediated, mechanisms of tissue damage may contribute to tissue damage of HSVE. PMID- 10457386 TI - Does selegiline modify the progression of early Parkinson's disease? Results from a five-year study. The Norwegian-Danish Study Group. AB - The objective of this study was to examine the effect of long-term treatment with selegiline on the progression of Parkinson's disease (PD). One hundred and sixty three patients with early PD were treated with levodopa and benserazide, combined with selegiline or placebo in a five-year randomized, placebo-controlled, double blind, parallel group study followed by a one-month wash-out of selegiline or placebo. The main outcome measures were assessments of the severity of parkinsonism, levodopa requirements and the development of end-of-dose motor fluctuations over time and after wash-out at the end of the study period. Results indicated that patients treated with the combination of selegiline and levodopa developed markedly less severe parkinsonism and required lower doses of levodopa during the five-year study period than patients treated with levodopa and placebo. There was no trend towards worsening during wash-out among patients previously treated with selegiline. The results cannot easily be explained by a symptomatic effect of selegiline. PMID- 10457387 TI - Different postural reaction patterns for expected and unexpected perturbations in patients with idiopathic Parkinson's disease and other parkinsonian syndromes. AB - Different postural reaction patterns after predictable and unpredictable perturbations during free stance were studied in 8 patients with idiopathic Parkinson's disease (iPD), in 4 patients with other parkinsonian syndromes (PS) and in 5 healthy controls. First, the amplitude of leaning maximally backward and forward was measured (condition I). Secondly, the body equilibrium was disturbed by self-paced, predictable, rapid arm elevations (condition II) and by sudden unpredictable toe-down and toe-up rotations of a supporting platform (condition III). Patients with PS particularly had difficulties in regaining body equilibrium after unexpected perturbations. In controls and patients with PS, unpredictable disturbances were better compensated in toe-down than in toe-up direction, whereas the opposite was true for patients with iPD. These results correspond to the fact that patients with PS had a specific leaning-backward impairment and patients with iPD, a leaning-forward impairment. The authors conclude that the differences in postural stability between patients with iPD and PS are caused by different pathophysiological mechanisms. These differences in postural stability could serve as an additional tool for differential diagnosis. PMID- 10457389 TI - Psychological approach of non-epileptic to epileptic patients: the dynamics of attitude changes during hospitalization. AB - Measures of interpersonal relations of non-epileptic patients to epileptic patients were studied in search of answers to two questions: what is the initial attitude of non-epileptic patients to epileptic patients; and, do any changes in attitude occur during time spent together in hospital? In order to study these personal relations, a formal analysis of preferences was carried out. Twenty-two non-epileptic patients admitted to the Neurology and Epileptology Department, Medical Centre for Postgraduate Education, Warsaw were studied. Subjects were asked to rank-order (from 1 to 9 points) suggested ways of distributing the profits of a hypothetical joint (e.g., with a room-mate) money-earning venture. The experimental procedure for each patient was repeated for all three room mates. Preferences were assessed three times - the day after admission to hospital, after 11 days and after 21 days in hospital. Following this procedure, it was possible to trace the dynamics of the patients' interpersonal relations. The data were correlated (Spearman's r(s)) and submitted to analysis of variance (MANOVA) with repeated measures. Analysis of the attitudes of patients with non epileptic neurological disorders towards epileptic patients revealed a dynamic tendency - from negative (measures one and partly two) to positive attitudes after three weeks spent together in hospital (measure three). PMID- 10457388 TI - EMG evidence of myopathy and the occurrence of titin autoantibodies in patients with myasthenia gravis. AB - We studied 65 patients with myasthenia gravis (MG). Clinical, neurophysiological, immunological, and histological findings suggested the coexistence of a presumed autoimmune myopathy. The clinical features were persistent pyridostigmine resistant weakness and atrophy of striated muscles. The myopathy was found more often in patients with late-onset MG than in those with early-onset (37% vs 13%). Patients with myopathy were also prone to have other immune disorders (47% vs 13%). Elevated titres of antibodies against titin were detected more often in patients with electromyography (EMG) evidence of myopathy than in the sera of those without, and only in late-onset MG cases. PMID- 10457390 TI - Neurological and electrophysiological examinations on three groups of workers with different levels of exposure to mercury vapors. AB - The authors performed neurological, visual evoked potentials (VEP) and electroneurography (ENG) examinations on three groups of workers with occupational exposure to mercury vapors (Hg(0)), and on a control group. The exposure of dental professionals (n = 36) was mild, that of chloralkali plant workers (n = 36) was intermediate, and that of workers from mercury works (n = 77) was very high. Symptoms and signs of micromercurialism were observed only in the group with the highest exposure to Hg(0). In comparison with the control group, a shortening of VEP latency and a decrease in amplitude were found in the exposed groups. The VEP changes correlated with Hg(0) excreted in urine after administration of a chelating agent - sodium 2,3-dimercapto-1-propan sulfonate (DMPS). The frequency of abnormal VEP results increased with increasing levels of exposure. ENG changes were observed only in the group with the highest exposure to Hg(0). An isolated decrease of sural nerve conduction velocity was observed in 18% of total workers. In 70% of the cases, this was associated with an abnormality in VEP. The combination of a decrease in sural nerve conduction velocity and an abnormality of VEP seems to be a characteristic pattern of electrophysiological changes in persons exposed to mercury vapors. PMID- 10457391 TI - Adhesion of mononuclear cells from multiple sclerosis patients to cerebral vessels in cryostat sections of normal human brain. AB - Leukocyte extravasation across the blood-brain barrier is a critical event in the pathogenesis of multiple sclerosis (MS). This complex multistep process includes the adhesion of leukocytes to the endothelial cells of the central nervous system microvasculature. To investigate this phenomenon in MS, we developed a modified version of the frozen-section assay. Peripheral blood mononuclear cells (PBM) from 26 MS patients, 26 healthy controls and 10 patients with other inflammatory non- neurological diseases (OIND) were co-incubated with cryostat sections of normal brain white matter, immunohistochemically labelled with anti-CD45 antibody and counterstained with Giemsa stain. CD45-positive PBM adherent to transected microvasculature were counted with an automated image analyzer. MS patients showed an increased number of vessel-bound PBM (48.8 +/- 36.4) with respect to healthy controls (27.4 +/- 20.7, P = 0.01) and OIND patients (22.6 +/- 7.8, P = 0.01). Significant differences were also obtained counting the number of vessel bound PBM as a percent of total vascular cells between MS patients (12.7 +/- 7.2%) and healthy controls (6.9 +/- 5.4%, P = 0.002) or OIND patients (7.4 +/- 4.4%, P = 0.03). We confirm that PBM from MS patients show an increased potential of binding to cerebral vessels. The frozen-section assay provides a unique tool to study in situ the molecular interactions of leukocytes with brain vascular structures. PMID- 10457392 TI - Increased intrathecal nitric oxide formation in multiple sclerosis; cerebrospinal fluid nitrite as activity marker. AB - Nitric oxide is formed from L-arginine by a family of enzymes: nitric oxide synthase (NOS). The inducible nitric oxide synthase is activated by cytokines and it has been suggested that activation of the enzyme gives rise to neurotoxic levels of reactive nitrogen oxides. This enzyme has been shown to be localized in multiple sclerosis (MS) lesions but the role of nitric oxide formation in the pathogenesis of MS is still unclear. Using capillary electrophoresis, we have analysed nitrite and nitrate in cerebrospinal fluid (CSF) and demonstrate increased levels of reactive nitrogen products in 17 patients with MS. The total levels of oxidized nitrogen products were significantly elevated in MS patients when compared with controls. In patients with active MS, nitrite levels were significantly increased when compared with controls and patients in remission. This is supportive of NOS induction in MS. We suggest that capillary electrophoresis analysis of nitrite and nitrate in CSF could provide a clinically useful way to determine disease activity in MS. PMID- 10457393 TI - Interleukin-12 is detectable in sera of patients with multiple sclerosis - association with chronic progressive disease course? AB - Multiple sclerosis (MS) is widely accepted as a systemic T- cell-mediated autoimmune disease with a T-helper type-1 (TH-1) profile of cytokine production. We addressed the question whether interleukin-12 (IL-12), as a central mediator of TH-1-cell activities, is detectable in sera of MS patients, and if there is any association with disease activity. We analysed 171 sera of patients with MS and meningitis, and healthy controls. IL-12 p40 protein was detectable at low levels in MS patients (median 43 pg/ml) and controls (median 49 pg/ml). Analysing different disease courses and activities, a significant elevation in stable primary progressive MS cases compared with controls (median 66 pg/ml) was found. IL-12 p40 protein was not detectable in cerebrospinal fluid probes of 10 patients. We conclude that the function of IL-12 in MS depends on expression and degradation of the different proteins. These could act proinflammatory as well as limiting the disease process. PMID- 10457395 TI - Features of dural sinus thrombosis simulating pseudotumor cerebri. AB - To characterize dural sinus thrombosis (DST) patients presenting with a syndrome simulating pseudotumor cerebri (PTC), medical records of patients initially diagnosed as having PTC between 1980 and 1995 were analysed. Inclusion criteria were intracranial hypertension (IH) and a normal initial computed tomography (CT) scan. All patients underwent angiography or magnetic resonance imaging (MRI) and magnetic resonance venography (MRV). Patients with IH secondary to a determined disorder were excluded from the study. Forty-six patients were included, 12 patients had MRI/MRV or angiography findings compatible with DST and the remaining 34 patients had no vascular pathology on neuroimaging studies and were considered to have PTC. No clinical or auxiliary findings differed between the two groups except for younger age, which delineated the PTC patients. Two patients with DST died during the study. Both were agitated, had a relatively rapid course of symptom progression, and a CSF pressure of over 40 cm water. We conclude that DST can be identified in 26% of patients presenting with symptoms and signs typical of PTC. It may be impossible to differentiate between patients with and without DST if MRI/MRV or angiography are not performed in this scenario. PMID- 10457394 TI - Cerebrospinal fluid filtration in amyotrophic lateral sclerosis. AB - By means of a randomized, controlled and open study the authors wanted to find out if cerebrospinal (CSF)-filtration was of substantial benefit to patients with sporadic amyotrophic lateral sclerosis (SALS). Five SALS patients, aged 51-75 years, being treated with riluzole underwent CSF-filtration daily over five days (group A). Five other SALS patients, aged 52-70 years, were treated only with riluzole (group B). Although all five patients in the first group reported a subjective benefit following CSF- filtration, the Norris score, the Frenchay score, the vital capacity, the ulnar nerve F-wave persistence and the peak-ratio of the brachial biceps and anterior tibial muscles did not change significantly after five days of therapy, either in group A or in group B. In conclusion, filtration of 200-250 ml CSF daily, over five days, does not seem to have a substantial therapeutic effect in patients with SALS. PMID- 10457396 TI - A novel mutation in the GTP cyclohydrolase I gene associated with a broad range of clinical presentations in a family with autosomal dominant dopa-responsive dystonia. AB - We examined a large family of Ashkenazi Jewish origin with autosomal dominant dopa-responsive dystonia (DRD). Mutation analysis of the GTP cyclohydrolase I gene revealed in affected members a novel point mutation (a C/A change in exon 1) resulting in a threonine-to-lysine substitution at residue 94. The mutation was characterized by variable expressivity and was associated with either a 'classical' DRD phenotype or various atypical phenotypes, such as subtle transitory equinovarus postures of the feet or isolated hand tremor. This observation demonstrates the significance of the molecular testing in establishing the clinical diagnosis of DRD. PMID- 10457397 TI - Trichloroethylene and parkinsonism: a human and experimental observation. AB - This report describes the case of a 47-year-old woman who developed Parkinson's disease after seven years of professional exposure to trichloroethylene. In the light of this clinical report, mice were intoxicated with trichloroethylene and tyrosine hydroxylase immunoreactivity was used to measure neuronal death in the substantia nigra pars compacta. Treated mice presented significant dopaminergic neuronal death in comparison with control mice (50%). The environmental trichlorethylene pollution, as well as other unspecific neurotoxic solvents, could potentially contribute to the genesis of some cases of Parkinson's disease. PMID- 10457399 TI - How safe is hydroxyurea in the treatment of polycythemia vera? PMID- 10457398 TI - Adult leigh syndrome: treatment with intravenous soybean oil for acute central respiratory failure. AB - This study reports a 38-year-old woman with adult Leigh syndrome associated with partial deficiency of the pyruvate dehydrogenase complex. The patient had intermittent diplopia, loss of vision, dystonia, central respiratory failure and unconsciousness with lactic acidosis. Treatment with an intravenous ketogenic emulsion resulted in rapid clinical and biochemical improvement. In patients with acute respiratory failure under these circumstances, intravenous ketogenic emulsion therapy is worth consideration. PMID- 10457400 TI - Autologous bone marrow transplantation for chronic myeloid leukemia. PMID- 10457401 TI - Short-term, serum-free, static culture of cord blood-derived CD34+ cells: effects of FLT3-L and MIP-1alpha on in vitro expansion of hematopoietic progenitor cells. AB - BACKGROUND AND OBJECTIVE: The use of ex vivo expanded cells has been suggested as a possible means to accelerate the speed of engraftment in cord blood (CB) transplantation. The aim of this study was to fix the optimal condition for the generation of committed progenitors without affecting the stem cell compartment. DESIGN AND METHODS: Analysis of the effects of FLT3-L and MIP-1alpha when combined with SCF, IL-3 and IL-6, in short-term (6 days), serum-free expansion cultures of CB-selected CD34+ cells. RESULTS: An important expansion was obtained that ranged between 8-15 times for CFU-GM, 21-51 times for the BFU-E/CFU-Mix population and 11 to 30 times for CD34+ cells assessed by flow cytometry. From the combinations tested, those in which FLT3-L was present had a significant increase in the expansion of committed progenitors, while the presence of MIP 1alpha had a detrimental effect on the generation of more differentiated cells. However, stem cell candidates assessed by week 5 CAFC assay could be maintained in culture when both MIP-1a and FLT3-L were present (up to 91% recovery). This culture system was also able to expand megakaryocytic precursors as determined by the co-expression of CD34 and CD61 antigens (45-70 times), in spite of the use of cytokines non-specific for the megakaryocytic lineage. INTERPRETATION AND CONCLUSIONS: The results obtained point to the combination of SCF, IL-3, IL-6, FLT3-L and MIP-1alpha as the best suited for a pre-clinical short-term serum-free static ex vivo expansion protocol of CB CD34+ cells, since it can generate large numbers of committed progenitor cells as well as maintaining week 5 CAFC. PMID- 10457402 TI - Peripheral T lymphocyte cytokine profile (IFNgamma, IL-2, IL-4) and CD30 expression/release during measles infection. AB - BACKGROUND AND OBJECTIVE: Measles virus infection (MVI) has been reported to be characterized by an imbalanced Th(1/2)-type cytokine profile. CD30 has been proposed as a receptor preferentially associated with the Th(0/2)-type cytokine pattern. The aim of this study was therefore to define the peripheral T lymphocyte cytokine profile and to test which CD30 expression pattern it was associated with in MVI. DESIGN AND METHODS: The design of the study was a prospective evaluation with comparative analysis. The serum levels of the soluble form of CD30 (sCD30) were determined at diagnosis and at weekly intervals up to 4 weeks, using an ELISA, in 23 males (median age 19), who developed MVI while serving in the Italian army and who were admitted to the Infectious Disease Unit of the Military Hospital in Padua. In 10 of the patients at diagnosis we studied the lymphoid immunophenotype and, after non-specific ex vivo stimulation, the expression of IFNgamma, IL-2 and IL-4 by peripheral T cells using flow cytometry single cell analysis. In 3 patients such evaluations were also performed 7 weeks later. RESULTS: At diagnosis, we found (i) reduction of IFNgamma+/CD4+ T cells (p=0.048 vs controls) in the absence of substantial variation of IL-2+ and IL-4+ T cells (p=ns vs controls); (ii) expansion of CD30+/ CD4+ and CD30+/CD8+ T cell subsets (p<0.01 vs controls); (iii) high sCD30 values (median 61 U/mL; p<0.001 vs controls); (iiii) a context of lymphopenia (0. 728+/-0.292 lymph x10(9)/L). sCD30 remained elevated up to 4 weeks from MVI onset [median values 53, 49, 50, 34 U/mL after 1, 2, 3 and 4 weeks, respectively (p=ns between different time points)]. In 3 patients tested 7 weeks after diagnosis, we still observed decreased IFNgamma production by CD4+ and CD8+ T cells (p=0.05 and <0.01, respectively vs controls) and reduction of CD4+ and CD8+/IL-2+ T cells (p<0.01). INTERPRETATION AND CONCLUSIONS: MVI was characterized by featuresof inadequate Th/Tc(1) activation associated with increased circulating CD30+ T cells and elevated sCD30 levels, supporting a correlation between Th/Tc status and CD30 expression/release pattern in vivo. PMID- 10457403 TI - 3q21 and 3q26 cytogenetic abnormalities in acute myeloblastic leukemia: biological and clinical features. AB - BACKGROUND AND OBJECTIVE: Acute myeloblastic leukemia (AML) with features of myelodysplastic syndrome (MDS) and abnormalities of megakaryocytopoiesis is often characterized by cytogenetic aberrations of the 3q21 and 3q26 bands involving inv(3)(q21q26) and (3;3)(q21;q26). These aberrations have been described in all FAB subtypes with the exception of M3, and in MDS and in megakaryoblastic crisis of chronic myeloid leukemia. We reviewed the biological and clinical features of 10 cases of AML with inv(3)(q21q26) and t(3;3)(q21;q26). DESIGN AND METHODS: Four hundred and sixteen patients with AML were studied in our Institute by cytogenetic analysis and 10 (2.4%) showed inv(3)(q21q26) (7 patients) or t(3;3)(q21;q26) (3 patients): 7 males, 3 females; median age, 43.5 yrs. We also used RT-PCR to investigate the pattern of expression of the EVI-1 gene in 5 patients. RESULTS: Additional chromosomal changes were demonstrated in 6 patients. In 5/10 cases a preceding MDS had been observed. A possible occupational exposure was established in 2 patients (a farmer and an histologist employing organic solvents) and another patient had a therapy-related leukemia. AML subtype was M1 in 9 patients and M2 in 1. A variable excess of micromegakaryocytes was observed in all the patients. In 5 patients the platelet count was normal or increased (median number: 172. 5x10(9)/L; range 55-440). Expression of EVI-1 gene was present in all the 5 patients studied. The clinical course and outcome was extremely poor: 9/10 patients were resistant and 1 patient showed a partial remission after induction therapy. Of the 9 patients resistant to the first line chemotherapy, 7 were also resistant to the second line chemotherapy. Three patients obtained a morphologic complete remission after third line chemotherapy (duration 1, 3 and 6 months); 2 of them were submitted to autologous bone marrow transplantation, but relapsed after 1 and 3 months. The median overall survival was 5.5 months. INTERPRETATION AND CONCLUSIONS: Our findings evidence a strong correlation between 3q21q26 chromosomal aberrations, abnormalities of megakaryocytopoiesis and lack of response to conventional chemotherapy and support the diagnostic and prognostic relevance of chromosome characterization in the classification of AML. PMID- 10457404 TI - Cleavage of the ALL1 gene in acute lymphoid leukemia before treatment disappears in relapse. AB - BACKGROUND AND OBJECTIVE: ALL1 gene rearrangements are frequently found in secondary acute leukemias (ALs). A site-specific cleavage of the ALL1 gene in a consensus sequence for topoisomerase II recognition has been considered to be the initial step leading to ALL1 rearrangement and subsequent therapy-related AL. The aim of the present study was to evaluate this cleavage in our patients, to analyze whether it is a laboratory-produced artefact and to check whether it persists or causes a real ALL1 gene rearrangement at relapse. DESIGN AND METHODS: We studied ALL1 rearrangement in 74 cases of AL before treatment by Southern blot avoiding room temperature exposure or delay in processing the samples which could produce ALL1 cleavage. DNA was available for two cases with ALL1 cleavage; it was analyzed by three different Southern blots in one and two in the other. One case with ALL1 cleavage was also studied in relapse. RESULTS: The presence of the cleavage of the ALL1 DNA was found in 3 of 74 (4%) patients. Two of these three patients had the ALL1 cleavage in three and two different analyses. One case was positive for ALL1 cleavage at diagnosis, but negative for both ALL1 cleavage and ALL1 rearrangement at relapse. INTERPRETATION AND CONCLUSIONS: The fact that a constant pattern was obtained from the same patients in different DNA preparations, supports the notion that ALL1 cleavage is not a laboratory artefact. The absence of the cleavage in a sample from a relapsed patient suggests that the subclone with the ALL1 cleavage, in this case, did not play a clear role in the pathogenesis of disease recurrence. PMID- 10457405 TI - Outcome of biphenotypic acute leukemia. AB - BACKGROUND AND OBJECTIVE: Although biphenotypic leukemia is now a defined entity, outcome of this rare form of acute leukemia has not been well documented. We present the first comprehensive study analyzing induction and consolidation therapy of biphenotypic leukemia and correlate outcome to prognostic factors. DESIGN AND METHODS: In this retrospective study, the incidence of biphenotypic leukemia was found to be 3.6% from 693 adult and pediatric acute leukemias referred to our center for treatment over the last 8 years. Of these, 15 were B lymphoid/myeloid, 8 were T-lymphoid/myeloid, one was T/B lymphoid and one had trilineage differentiation. RESULTS: Induction of remission in de novo cases was achieved in 70% of patients and relapse of disease occurred in 15%. The use of combined lymphoid and myeloid drugs for induction resulted in a high incidence of early deaths (25%). The overall probability of survival at 2 years was 39.4%. Patients with secondary disease had a uniformly poor outcome with low remission rates and high relapse rates. INTERPRETATION AND CONCLUSIONS: Prognosis was most strongly related to the presence of the Philadelphia chromosome (p=0.03) and age under 15 years (p=0.01). We conclude that patients with biphenotypic leukemia should have risk stratification with treatment tailored to their prognostic factors. PMID- 10457406 TI - A prospective study of alpha-interferon and autologous bone marrow transplantation in chronic myeloid leukemia. The Italian Co-operative Study Group on Chronic Myeloid Leukemia. AB - BACKGROUND AND OBJECTIVE: Alpha-interferon (alphaIFN) can induce cytogenetic remissions in chronic myeloid leukemia (CML). Hemopoietic progenitors can be collected from the marrow in remission and utilized for autologous repopulation after high dose chemotherapy. This study was designed with the purpose of evaluating the feasibility of a combined treatment policy of alphaIFN followed by autologous bone marrow transplantation (autoBMT). DESIGN AND METHODS: A prospective study of alphaIFN and autoBMT was begun in 1989. Two hundred and seventy-two consecutive previously untreated non-blastic Ph positive chronic myeloid leukemia (CML) patients, who were less than 56 years old, were enrolled over a 3-year period (1989-1991) and were assigned to receive human recombinant alphaIFN 2a (Roferon-A) at a dose of 9 MIU daily for at least one year. If they achieved a cytogenetic response consisting in a percentage of Ph neg metaphases of more than 25%, they were eligible for marrow harvesting and subsequent autografting after high dose busulfan (16 mg/kg) and melphalan (60 mg/m(2)). RESULTS: Seventy-six patients (28%) were eligible for a marrow harvest but the marrow was harvested in only 37 cases (14%), and only twenty-three patients (8%) were actually autografted. One patient died of infection nine days after autoBMT. The other patients recovered and did not suffer any late adverse events. Five patients progressed to blastic phase, six are alive in complete hematologic remission and eleven are alive in complete hematologic and cytogenetic remission. AlphaIFN treatment was reinstituted after autoBMT in 18 of 22 cases, but four patients who are in continuous complete cytogenetic remission were not given alphaIFN anymore. The progression-free survival of the autografted patients is 65% 8 years after registration. INTERPRETATION AND CONCLUSIONS: This study shows that bone marrow hemopoietic progenitors (Ph neg and Ph pos) can be collected from patients who respond to alphaIFN and can be used to rescue hemopoietic activity after high dose chemotherapy. Though some complete and durable cytogenetic remissions were obtained, the treatment could be applied only to a small group of good risk patients, highlighting that selection is very important and results cannot be extrapolated to the average patient. PMID- 10457407 TI - Favorable impact of low-dose fludarabine plus epirubicin and cyclophosphamide regimen (FLEC) as treatment for low-grade non-Hodgkin's lymphomas. AB - BACKGROUND AND OBJECTIVE: In recent years, conventional dose of fludarabine (FLU) alone or in combination with other drugs has been reported to be effective in the treatment of low-grade non-Hodgkin's lymphomas (LG-NHL). In particular, FLU and cyclophosphamide (CY) or FLU and mitoxantrone or idarubicin combined regimens have shown considerable therapeutic activity both as first line and salvage therapies, producing overall response rates ranging from 40-50% in previously treated patients and up to 70-90% in untreated ones. However severe neutropenia and infective complications have been reported in a significant number of patients. Based on these premises we evaluated the efficacy and toxicity of a new regimen combining low-doses of FLU with epirubicin (EPI) and CY (FLEC) in a group of advanced treatment-requiring LG-NHL patients. The aim of this study was to evaluate a strategy aimed at lowering therapy-related toxic effects without affecting the reported good response rate. DESIGN AND METHODS: Thirty patients with de novo, relapsed or refractory LG-NHL entered the study. FLEC regimen was as follows: EPI 60 mg/m(2) i.v. on day one, plus FLU 15 mg/m(2)/day i.v. (max 25 mg) and CY 250 mg/m(2)/day i.v. for four days. RESULTS: All 30 patients were evaluable for response, 13 (43%) fulfilled the criteria for CR and 11 (36%) for PR with an overall response rate of 79%. None of the 13 patients who achieved CR had relapsed after a follow-up of 2 to 23 months (median duration 13 months). With regard to age, 13/14 older patients (>/= 70 years) responded to the treatment and 9 of them maintained their response after a median of 13 months (range 2-22); six of the 14 (43%) obtained a CR. Therapy-related toxicity was mild regardless of age, neutropenia (43%) and fever of undetermined origin (26%) being the major side effects. Remarkably, a documented infection was recorded only in 2/30 (6%) patients. INTERPRETATION AND CONCLUSIONS: A low-dose FLU-based FLEC regimen appeared to be effective for advanced treatment-requiring LG-NHL, reproducing a similar overall response rate (79%) reported to have been achieved with other FLU based combination therapies. Toxic side effects were negligible and in particular documented infections were remarkably uncommon even in the group of elderly patients. PMID- 10457408 TI - Effect of adenosine derivatives on in vitro thrombus formation induced by shear stress. AB - BACKGROUND AND OBJECTIVE: Shear-stress is considered to be the first event of platelet aggregation in vivo and platelet adhesion may be enhanced under pathologic conditions (e.g. arterial occlusion). DESIGN AND METHODS: We wanted to test the effect of adenosine derivatives on platelet aggregation induced by shear stress. By increasing platelet cAMP adenosine derivatives inhibit platelet activation. This in turn leads to P-selectin (CD62P) exposure, which is known to play a fundamental role in the binding of platelets to leukocytes. This gives rise to thrombus formation. RESULTS: The levels of cAMP (pmol/mL) prior to and after treating blood with the following compounds were respectively: PGE1 4.67+/ 0.29, 9.33+/-0. 58; SP64 5.2+/-0.34, 6.83+/-0.52; 2-Cl-adenosine 5.83+/- 0.58, 7. 45+/-0.55; NECA 7.00+/-2.29, 8.00+/-1.76. INTERPRETATION AND CONCLUSIONS: High shear rate was studied using a filteraggregometer which could be a good test for analyzing what happens under physiologic conditions compared to other systems in which platelet aggregation only occurs after adding aggregating agents (Born aggregometer). PMID- 10457409 TI - Lipoprotein(a) concentration is not associated with venous thromboembolism in a case control study. AB - BACKGROUND AND OBJECTIVE: Lipoprotein(a) is an LDL-like particle displaying strong athero-thrombotic properties. Although Lp(a) plays a pivotal role in the genesis and progression of thrombosis in the arterial district, its role in promoting thrombosis in the venous district is still unclear. DESIGN AND METHODS: To give further insight into the thrombotic potential of Lp(a), 100 potentially eligible consecutive outpatients who had suffered from previous episodes of venous thrombosis (deep vein thrombosis with or without pulmonary embolism) were enrolled into the study. Thirty-six of these patients who did not fulfil the entry criteria were then excluded from the study. The concentration of Lp(a) was thus measured in 64 patients, and compared to that of 64 control subjects, matched for sex (p=0.46), age (p=0.25) and pharmacological treatment; no subject belonging to the control group had a familial or personal history of venous thromboembolism. Exclusion criteria for both groups included: diabetes mellitus, liver or kidney diseases and malignancy, as established by both laboratory analysis and physical examination. To rule out false elevations of Lp(a) due to the presence of a concurrent acute phase response, C reactive protein (CRP) was measured in both groups using a commercial immunonephelometric assay. RESULTS: No statistically significant differences were observed in the median Lp(a) concentration between patients and controls (median: 69 vs 83 mg/L, respectively; p=0.34). Neither were any significant differences found between patients who had suffered from deep venous thrombosis with (n=18) or without (n=46) pulmonary embolism (median: 73 vs 69 mg/L, respectively; p=0. 83). The concentration of CRP did not differ significantly between cases and controls (median: 1.8 vs 2.3 g/L, respectively; p=0.37). INTERPRETATION AND CONCLUSIONS: Although there are several plausible biological mechanisms to explain the strong thrombogenicity of Lp(a) in vitro, we failed to demonstrate a convincing association between Lp(a) and thrombosis in the venous district. Besides the proven prothrombotic role of Lp(a) in some selected clinical settings, it is thus conceivable that the contribution of Lp(a) to genesis and progression of the venous thrombosis might be marginal or efficiently counterbalanced in vivo. The clinical usefulness of including the measurement of Lp(a) among the screening tests for thrombophilic patients, therefore, remains questionable. PMID- 10457410 TI - Placental/umbilical cord blood transplantation. AB - In this article we summarize the clinical outcome of unrelated placental/umbilical cord blood (CB) transplantation, discuss the biological characteristics of CB hematopoietic progenitor/stem cells (HPC) and balance the relative advantages and disadvantages of this therapy as compared with transplantation of other HPC sources. Moreover, we discuss CB banking programs at local, national and international levels. The data reported by the investigators of the New York Placental/Umbilical Cord Blood Program and of the Eurocord group indicate that the clinical outcome of allogeneic unrelated CB transplantation is significantly related to cell dose, being more effective in children than in adults, and is highly dependent on disease stage at transplantation. Furthermore, both studies show lower graft-versus-host disease (GvHD) frequency and severity and prolonged time intervals for platelet engraftment as compared to those of bone marrow and mobilized peripheral blood recipients. Although the data from the New York Placental/Umbilical Cord Blood Program seem to support a negative effect of HLA differences, the latter were not significantly associated with survival in the Eurocord series. Additional observations are therefore necessary to collect conclusive evidence in this regard. Currently available data show that CB contains a higher proportion of primitive HPC and that CB-HPC possess higher proliferation and expansion potentials as compared to adult bone marrow. Furthermore, there is some evidence indicating that CB-HPC are more adequate than HPC from other sources for genetic manipulation and gene therapy. Despite the significant advances in the knowledge of the biology and in the clinical use of CB, a number of problems remain unsolved, including the standardization of banking procedures and unit quality and the development of suitable protocols for transplantation of adult patients. PMID- 10457411 TI - Homozygous form of the Pelger-Huet anomaly. PMID- 10457412 TI - Bilobulated circulating lymphocytes in persistent polyclonal B-cell lymphocytosis. PMID- 10457413 TI - Atypical clinical presentation of visceral leishmaniasis. PMID- 10457414 TI - Immunosuppression due to MACOP-B does not seem to cure the antiphospholipid syndrome. PMID- 10457415 TI - New technology and changing parameters of leukapheresis for blood cell transplantation. PMID- 10457416 TI - INR variability in anticoagulation with acenocoumarol: is it useful for identifying patients at risk of bleeding and thrombosis? PMID- 10457417 TI - The frequency of allele alpha(LELY), a low expression allele of the gene encoding erythroid spectrin alpha-chain, in the Greek population. PMID- 10457418 TI - Acute myeloid leukemia occurring in a patient with polycythemia vera in treatment with hydroxyurea. PMID- 10457419 TI - Absence of p53 mutation in 15 cases of myeloid malignancies with structural rearrangements of 3q. PMID- 10457420 TI - Invasive cerebral aspergillosis in a patient with aplastic anemia. Response to liposomal amphotericin and surgery. PMID- 10457421 TI - Transient response of myeloma clone to pamidronate therapy. PMID- 10457422 TI - A paradoxical side-effect of antiaggregating treatment with ticlopidine: the Moschowitz syndrome. PMID- 10457423 TI - Flow cytometry of cell suspensions from lymph nodes: immunophenotype, DNA content and proliferative rate are strongly correlated with histopathology diagnosis. PMID- 10457424 TI - Previous occurrence of life-threatening abdominal infection is not a contraindication to bone marrow transplantation. PMID- 10457425 TI - Splenic inflammatory pseudotumor mimicking primary splenic malignancy. PMID- 10457426 TI - Concomitant chronic lymphocytic leukemia and acute myeloid leukemia diagnosed by two color flow cytometric analysis. PMID- 10457427 TI - Acute megaloblastic anemia: homocysteine levels are useful for diagnosis and follow-up. PMID- 10457429 TI - Chromatographic resolution of the enantiomers of a pharmaceutical intermediate from the milligram to the kilogram scale. AB - The preparative chromatographic resolution of racemic mixtures is rapidly becoming a standard approach for the generation of enantiomers in pharmaceutical R&D. This paper will discuss the optical resolution of a pharmaceutical intermediate as the separation is scaled up from the milligram to the kilogram scale. Difficulties encountered and their solutions at each scale will be discussed. In addition, the exploration of Simulated Moving Bed (SMB) for the separation will also be discussed. Finally, a comparison of the productivities and solvent consumption for each method and scale will be presented. PMID- 10457428 TI - Radiation immobilization of poly(methyloctylsiloxane) on silica for use in HPLC: a uniform layer model. AB - Poly(methyloctylsiloxane) (PMOS) was sorbed into the pores of HPLC silica by a solvent evaporation procedure, then irradiated with gamma rays from a cobalt-60 source to absorbed doses in the range from 0 to 200 kGy (1Gy = 1J kg-1). Non irradiated and irradiated samples were characterized by solvent extraction, specific surface area determination, infrared spectroscopy and reversed-phase column performance. Solvent extraction data reveal that about 40% of the PMOS is not extractable prior to irradiation and this increases to about 75% with radiation doses of 50 kGy or higher. Column performance was improved by the radiation treatment, reaching a maximum efficiency in the dose range of 80-140 kGy while the peak symmetry changed from As = 1.7 to 1.1. The improvement is attributed to the increased mass of polymer immobilized by the radiation treatment and to a more uniform distribution of the immobilized polymer in the silica pore system. A multi-layer stationary-phase model is presented in which the first layer consists of an adsorbed monolayer of PMOS and the second layer is immobilized by gamma radiation. PMID- 10457430 TI - Molecularly imprinted adsorbents for positional isomer separation. AB - 2,4-Dihydroxybenzophenone (2,4-DHB) imprinted polymers were synthesized by surface imprinting technique, using allyl alcohol as the functional monomer. The polymers showed a very high selectivity for 2,4-DHB when compared with various positional isomers such as 2-HB, 2,2'-DHB, 4,4'-DHIB and 4,4'-DMB. Solvents were found to affect the selectivity as well as sorption capacity in the case of surface imprinted polymers. The selectivities decreased drastically when the imprint cavity was blocked. This validated the importance of the cavity and the rebinding interactions in governing the selectivity in the case of MIPs. The surface imprinted polymers also showed a high selectivity under non-equilibrium conditions thereby making them suitable adsorbents for industrial separations. PMID- 10457431 TI - Uniform-sized molecularly imprinted polymer for (S)-naproxen selectively modified with hydrophilic external layer. AB - A uniform-sized molecularly imprinted polymer (MIP) for (S)-naproxen selectively modified with hydrophilic external layer has been prepared. First, the molecularly imprinted polymer for (S)-naproxen was prepared using 4-vinylpyridine and ethylene glycol dimethacrylate (EDMA) as a functional monomer and cross linker, respectively, by a multi-step swelling and thermal polymerization method. Next, a 1:1 mixture of glycerol monomethacrylate (GMMA) and glycerol dimethacrylate (GDMA) was used for hydrophilic surface modification, and it was added directly to the molecularly imprinted polymer for (S)-naproxen 4 h after the start of molecular imprinting. The retention factors of all solutes tested were decreased with the surface modified molecularly imprinted polymer, compared with the unmodified molecularly imprinted polymer. However, chiral recognition of racemic naproxen was attained with the surface modified molecularly imprinted polymer as well as the unmodified molecularly imprinted polymer. Further, bovine serum albumin was completely recovered from the surface modified molecularly imprinted polymer. These results revealed that the chiral recognition sites of (S)-naproxen remained unchanged with hydrophilic surface modification, and that the molecularly imprinted polymer for (S)-naproxen was selectively modified with hydrophilic external layer. Preliminary results reveal that the surface modified molecularly imprinted polymer could be applicable to direct serum injection assays of (S)-naproxen. PMID- 10457432 TI - Determination of selected phytochemicals by reversed-phase high-performance liquid chromatography combined with ultraviolet and mass spectrometric detection. AB - A robust, routinely manageable and sensitive RP-HPLC method combined with UV (270 nm) and ESI-MS detection was established for the determination of abundant pertinent phenolic compounds (phytochemicals) from various biological matrices. Phytochemicals were extracted by aqueous methanol (80%), extracts were analysed without further purification. Baseline separation was achieved within 30 min for 19 phytochemicals and excellent sensitivity (6-42 pmol at S/N = 3) was obtained. The identity of the phytochemicals was confirmed with standard compounds and with LC-MS. The repeatabilities for the majority of the phytochemicals ranged between 3% and 6%. The practicability of the method was shown in complex biological matrices by analysing onion and soybean extracts. This generally applicable technique may serve as a valuable tool for a rapid screening and a specific measurement of phytochemicals in food extracts and biological fluids and serve as analytical instrument for future biochemical and physiological studies. PMID- 10457433 TI - Simultaneous quantification of five major biologically active ingredients of saffron by high-performance liquid chromatography. AB - A simple, sensitive and specific high-performance liquid chromatography-UV (HPLC UV) method has been developed for the first time to simultaneously quantify the five major biologically active ingredients of saffron, namely crocin 1, crocin 2, crocin 3, crocin 4 and crocetin. Calibration curves were derived by spiking authentic compounds and internal standard, 13-cis-retinoic acid, into herbal samples prior to extraction. Extraction was conducted simply by stirring dried herb (20 mg) with 80% aqueous methanol (5 ml) at ambient temperature in the dark for 2 h. The HPLC assay was performed on a reversed-phase C18 column with linear gradient elution using methanol and 1% aqueous acetic acid. Calibrations were linear (r2 = 0.999) for all five analytes, with overall intra- and inter-day RSDs of less than 11%. The assay was successfully applied to the determination of four crocins and crocetin in three saffron samples and two Zhizi, another crocin containing herb. Results indicate that the developed HPLC assay can be readily utilized as a quality control method for crocin-containing medicinal herbs. PMID- 10457434 TI - Determination of fatty acids in vegetable oil by reversed-phase liquid chromatography with fluorescence detection. AB - The effect of temperature and organic solvent composition (acetonitrile and methanol) on the reversed-phase separation of coumarin-derivatized fatty acids according to their carbon number (C14 to C22), the degree of unsaturation, as well as cis/trans (C18:1 c/t, C18:2 cc/tt, C18:3 ccc/ttt) configuration was investigated to find out the effective separation condition. Based on the linear plots of the logarithm of the capacity factor of saturated fatty acids versus their carbon number, the equivalent chain length (ECL) of unsaturated fatty acids was calculated. The ECL values were found to be significantly altered and the differentiation between cis and trans fatty acids was increased when either the temperature or organic solvent composition was decreased. These results generally led to a better resolution at the expense of separation time. A ternary gradient composed of water, acetonitrile, and methanol was then developed to elute the solutes at 55 degrees C within a separation time of 40 min with a minimum resolution of 1.0 for the worst pair. This method was demonstrated to resolve the fatty acids in a vegetable shortening. PMID- 10457435 TI - Improvement in the Iatroscan thin-layer chromatographic-flame ionisation detection analysis of marine lipids. Separation and quantitation of monoacylglycerols and diacylglycerols in standards and natural samples. AB - Mono- and diacylglycerols are important intermediates in glycerolipid biodegradation and intracellular signalling pathways. A method for mass determination of these lipid classes in marine particles was developed using the Iatroscan, which combines thin layer chromatography (TLC) and flame ionisation detection (FID) techniques. We improved existing protocols by adding two elution steps: hexane-diethyl-ether-formic acid (70:30:0.2, v/v/v) after triacylglycerol and free fatty acid scan, and acetone 100% followed by chloroform-acetone-formic acid (99:1:0.2, v/v/v) after 1,2 diacylglycerols. Diacylglycerol isomers 1,2 and 1,3 were separated from each other, as well as from free sterols in standards and marine lipids from sediment trap particles. Monoacylglycerols were separated from pigments and galactosyl-lipids in the same trap samples and in a rich pigment phytoplankton extract of Dunaliella viridis. Quantitation of each class in samples was performed after calibration with 0.5 to 2 micrograms of standards. As many as 17 lipid classes can be identified and quantified in samples using this proposed six-step development. PMID- 10457436 TI - Improvement of EPA method 8330: complete separation using a two-phase approach. AB - Complete separation of the 14 nitroaromatic and nitramine analytes targeted in EPA Method 8330 was achieved using a 30 x 4.6 mm Bondesil CN guard column in series with a 250 x 4.6 mm Bondesil C18 column (5 microns particles). Consistent with Method 8330 specifications, the mobile phase in the separation was 1:1 methanol:water and the flow-rate was 1.5 ml min-1. The success of this two-phase system proved to be largely a consequence of the unique resolution afforded by the Bondesil C18 column: complete separation of the 14 explosives could not be achieved using other CN/C18 column combinations. Additionally, while Method 8330 calls for a 100-microliter injection loop, separation on the Bondesil CN/C18 system was possible only using a 20-microliter injection loop. The loss of resolution with larger injection volume appears to be a result of the injection solvent, methanol, modifying the composition of the mobile phase both in the CN guard column and in the initial portion of the C18 column. The current method nevertheless represents an improvement over Method 8330, which calls for both a screening and confirming analysis. By separating the 14 Method 8330 analytes in a single run, the two-phase approach can increase sample throughput by decreasing analysis times. PMID- 10457437 TI - Separation of N-nitrosoamino acids by C18 reversed-phase ion-pair high performance liquid chromatography and compatible detection by electrospray ionization mass spectrometry. AB - Four non-volatile N-nitrosoamino acids, namely N-nitrososarcosine, N nitrosoproline, N-nitrosothiazolidine-4-carboxylic acid and N-nitroso-2-methyl thiazolidine-4-carboxylic acid were separated by C18 reversed-phase ion-pair high performance liquid chromatography (HPLC) using 1.4 mM C16-cetyltrimethylammonium chloride in methanol-water-acetonitrile (60:35:5, v/v) as the mobile phase. The N nitrosoamino acids were sensitively detected by negative electrospray ionization mass spectrometry (ESI-MS) in the form of the deprotonated carboxylate anion, [M H]-. Compatibility problems associated with HPLC separation and ESI-MS detection, such as formation of solvent cluster ions and the effects of co-eluting anions of the ion-pairing reagent, were systematically investigated. The optimized experimental conditions for separation and detection of N-nitrosoamino acids were described. PMID- 10457438 TI - Comparative studies on chemically and enzymatically coupled DNA-Sepharose columns for purification of a lac repressor chimeric fusion protein. AB - The length of a DNA sequence attached to an affinity chromatography column affects column retention of transcription factors. Even when unrelated sequences such as a poly(A):poly(T) tail are included in a DNA sequence, transcription factors such as the lac repressor are bound more tightly by the column. The position of the additional sequences is also important. To compare coupling procedures, an identical DNA sequence was covalently attached to Sepharose by chemical coupling or produced enzymatically by template driven enzymatic primer extension. These two types of supports, containing the O1 operator sequence bound by lac repressor, were packed into identical columns and compared by purification of a lac repressor-beta-galactosidase fusion protein. We found that the purity and yield of proteins eluted from the two columns were similar. Overall, the results suggest that there is no significant advantage to either type of support for the purification of some proteins. The study revealed a potentially important effect of the length of DNA sequences on column selectivity. PMID- 10457439 TI - Hydrophobic interaction chromatography of Chromobacterium viscosum lipase on polypropylene glycol immobilised on Sepharose. AB - The fractionation of Chromobacterium viscosum lipase was performed using a polypropylene glycol-Sepharose gel. The influence of mobile phase composition on the adsorption of lipase on the gel was studied and it was found that the retention of lipase depends on the salt used and increased with increasing the ionic strength. The retention was not strongly affected by changing the pH value of the mobile phase. By using 20% (w/v) ammonium sulphate in phosphate buffer a total retention of lipase on the column was obtained and by simply decreasing the ionic strength of the buffer, desorption of lipase could be achieved. The chromatographic purification of Chromobacterium viscosum lipase by hydrophobic interaction chromatography on Sepharose CL-6B modified by covalent immobilisation of 1,4-butanediol diglycidyl ether, polyethylene glycol and polypropylene glycol was also compared. PMID- 10457440 TI - Isolation of indole alkaloids from Catharanthus roseus by centrifugal partition chromatography in the pH-zone refining mode. AB - Centrifugal partition chromatography (CPC) in the pH-zone refining mode allowed a preparative and efficient isolation of vindoline, vindolinine, catharanthine and vincaleukoblastine from a crude mixture of Catharanthus roseus alkaloids. The separation protocol was tested with a synthetic mixture of vindoline, catharanthine and vincaleukoblastine. The fraction content was analyzed and the results compared with theoretical chromatograms obtained by numerical simulation. The increase in injected sample mass results in an improvement of the purity of the isolated compounds. This observation, confirmed by theory, is of prime importance for the development of preparative pH-zone refining CPC as a preparative separation method. PMID- 10457441 TI - Determination of the sulfur mustard hydrolysis product thiodiglycol by microcolumn liquid chromatography coupled on-line with sulfur flame photometric detection using large-volume injections and peak compression. AB - A selective, direct and relatively rapid method has been developed for the determination of thiodiglycol (TDG) in aqueous samples. TDG is the main hydrolysis product of the chemical warfare agent sulfur mustard. The method of analysis is based on the on-line coupling of reversed-phase microcolumn liquid chromatography and sulfur-selective flame photometric detection. To improve sensitivity and efficiency, peak compression by displacement was used in combination with large-volume injections. A concentration of 1% n-propanol was added to the sample to obtain the best sensitivity and efficiency after a 10 microliters injection. Detection limits of 0.25 microgram/ml were achieved with efficiencies of 4.10(5) plates per meter. The method was successfully applied during the Fourth Official Proficiency Test organized by the Technical Secretariat of the Organization for Prohibition of Chemical Weapons for the determination of TDG in a soil sample. PMID- 10457442 TI - Kinetic study of the mass transfer of S-Troger's base in the system cellulose triacetate and ethanol. AB - A recent study of the mass transfer kinetics of (-)- or S-Troger's base (TB) between ethanol and microcrystalline cellulose triacetate (CTA) allows an analysis of the concentration dependence of the mass transfer rate coefficient (k(m)). S-TB elutes before R-TB. The retention time of the both compounds decreases with increasing temperature. In this study, experimental data measured between 30 and 50 degrees C were analyzed to provide information on the kinetics of several mass transfer processes which take place in the chromatographic column, i.e., axial and intraparticle dispersion, the fluid-to-particle mass transfer, and the kinetics of adsorption/desorption at the actual adsorption sites. Intraparticle diffusion has the dominant contribution to band broadening at high flow-rates. Both intraparticle diffusivity and the surface diffusion coefficient exhibit a small concentration dependence. The positive dependence of k(m) on the concentration of S-TB seems to result from the properties of the adsorption/desorption kinetics and can be interpreted by considering the phase equilibrium properties. A quantitative analysis of the activation energy of the mass transfer kinetics of S-TB in the CTA column was also attempted. PMID- 10457443 TI - Quantitative determination of nonylphenol polyethoxylate surfactants in marine sediment using normal-phase liquid chromatography-electrospray mass spectrometry. AB - A new comprehensive analytical method based on normal-phase liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) has been developed for the quantitative determination of individual nonylphenol ethoxylate (NPEO) surfactants in complex environmental matrices. Clean-up of sample extracts was performed on cyanopropyl silica solid-phase extraction cartridges. Complete NPEO oligomer separation was achieved by using normal-phase LC. Because the non-polar solvents used in normal-phase LC are incompatible with ESI, unique LC-ESI-MS interface conditions were adopted that provided a functional interface and also enhanced the detection response of NPEOs. These provided enhanced ESI signal intensity and stability and facilitated the detection of NPEOs as sodium adducts at parts-per-billion concentration levels. The overall analytical method was validated for accuracy and precision by analyzing sediment samples spiked with known amounts of NPEOs. The method is superior to those currently used for NPEO analysis (LC-UV, LC-fluorescence, LC-thermospray-MS, LC-field desorption-MS, LC particle beam-MS and GC-MS) in terms of detection limits, specificity and speed of analysis. The validated method was successfully applied to determine levels of NPEOs in sediments from the Strait of Georgia, British Columbia. This work also demonstrates that by proper selection of normal-phase LC-ESI-MS interface conditions this technique is capable of solving separation problems which are not amenable with reversed-phase LC-ESI-MS. PMID- 10457444 TI - Development of an on-line coupling of liquid-liquid extraction, normal-phase liquid chromatography and high-resolution gas chromatography producing an analytical marker for the prediction of mutagenicity and carcinogenicity of bitumen and bitumen fumes. AB - A fast and fully automated system for the determination of polycyclic aromatic compounds (PACs) is described. The system has been developed to produce an analytical 'marker', correlating chemical characteristics (including PAC analysis) with mutagenicity and carcinogenicity. The products of interest are bitumen fumes, bitumen and other (heavy or even residual) oil products, regardless of their boiling range. Dimethyl sulphoxide (DMSO) extractables obtained from a flow-injection analysis (FIA) system are introduced on-line in a normal-phase liquid chromatographic (NPLC) system. Here, the PACs are separated from the DMSO and possible co-extracted heavy residual species. The final step incorporates on-line gas chromatographic analysis of the three-to-six-ring PAC fraction, followed by flame-ionisation detection for quantification. It was demonstrated that data obtained from samples in the distillate lubrication-oil range correlate well with data obtained from the manual DMSO-extraction method standardised by the Institute of Petroleum as IP346. PMID- 10457445 TI - Semi-empirical topological method for the prediction of the chromatographic retention of cis- and trans-alkene isomers and alkanes. AB - A new index is proposed for the prediction of the chromatographic retention of the cis- and trans-n-alkene isomers and alkanes. This index is based on the hypothesis that the chromatographic retention of the molecule is due to the interaction of each carbon atom with the stationary phase, and consequently the index is reduced by its neighbours' steric effects. The topological values are obtained by a numerical approximation considering the general behaviour of the chromatographic retention of the compounds. The simple linear regressions between the chromatographic retention and the index proposed for all branched alkanes and also isomers of the studied straight-chain C5 and C14 alkenes (1-ene, cis- and trans-2-, 3-, 4-, 5-, 6- and 7-enes) is very good (the correlation coefficient is r = 0.9999), and the elution sequence is correct for most of them. The models have a high predictive ability, as established by cross-validation values (r2cv). Thus, this new method, different from those already existent, can be used as complementary tool for the elucidation of the molecular structure, or prediction of the chromatographic retention of the cis- and trans-alkene isomers and branched alkanes. It could be extended with success, in the future, to the other types of compounds. PMID- 10457446 TI - Monitoring the synthesis of new pitches from coal tar and its fractions by chromatography and related techniques. AB - The aim of this paper is to study the applications of chromatography and related techniques in the transformation of coal-derived products into pitches for specific uses. Anthracene oil, tar and pitch were thermally treated in the presence of air (and sulfur in the case of anthracene oil) in order to cause the polymerization/condensation of their components. The evolution of the components of the parent materials during the treatments was monitored by gas chromatography mass spectrometry, gas chromatography-atomic emission detection, probe mass spectrometry and size-exclusion chromatography. From the results obtained, possible mechanisms for the transformation of coal-derived products into new pitches were established. PMID- 10457447 TI - Analysis of odorous compounds in water by isolation by closed-loop stripping with a multichannel silicone rubber trap followed by gas chromatography-mass spectrometry. AB - An alternative technique for the isolation and concentration of odorous compounds found in potable water is described. The method currently employed by water authorities is closed-loop stripping with the collection of these substances on a small activated carbon filter. The compounds of interest are then extracted from the carbon using a suitable solvent. The authors offer a multichannel silicone rubber trap as an alternative to the carbon filter. The absorbed compounds are thermally desorbed from the trap, directly on to the gas chromatographic column for analysis by GC-MS, thereby eliminating the solvent extraction step required by the carbon filter. The multichannel silicone rubber trap, producing equivalent results, offers a number of advantages over the carbon filter. PMID- 10457448 TI - Detection of the chemical warfare agents bis-(2-chloroethyl)ethylamine (HN-1) and tris-(2-chloroethyl)amine (HN-3) in air. AB - This paper describes the method development and validation for detection of the chemical warfare agents HN-1 and HN-3 in air using C8 solid-phase extraction disks followed by liquid desorption and analysis by gas chromatography. The method is contrasted to the standard approach which uses solid sorbent tubes followed by thermal desorption and analysis by gas chromatography. PMID- 10457449 TI - Determination of fatty acid amides as trimethylsilyl derivatives by gas chromatography with mass spectrometric detection. AB - Fatty acid amides are a newly emerging class of compounds with biological activity. The amides are formed enzymatically in vivo. Analysis of fatty acid amides has been accomplished by gas chromatography coupled with mass spectrometry. Fatty acid amides required derivatization prior to analysis at high temperatures due to thermal instability. Trimethylsilylation of fatty acid amides has been accomplished under optimum reaction conditions. The limit of detection for the silylated amides is approximately 1 pmol, with the lowest detected level being 700 fmol for the lauramide derivative. Quantitation of fatty acid amide derivatives can be accomplished by monitoring m/z 59 or m/z M-71, the only two major fragments formed in the ion trap mass spectrometer with electron impact ionization. The smaller fragment is the result of a newly reported, McLafferty type rearrangement; M-71 resulted from loss of an n-pentyl fragment. Either peak gave four-five orders of magnitude linear dynamic range. Numerous trimethylsilylamides from C7 to C20 were separated under standard conditions. Elution was linear with the number of carbons and was systematically affected by the number and position of the double bonds. PMID- 10457450 TI - Capillary gas chromatography coupled with microplasma mass spectrometry for organotin speciation. AB - Gas chromatography was coupled with microplasma mass spectrometry for selective detection of organotin compounds. The microplasma ion source was a capacitively coupled radiofrequency helium plasma, which was located inside the high vacuum area of the mass spectrometer. Only 1-3 ml min-1 of helium carrier gas from the gas chromatograph was necessary for sustaining the plasma while 0.15-1.5 ml min-1 of hydrogen was added as reagent gas. Hydrogen was applied for prevention of carbon deposition and served to minimize the interactions between tin and the fused-silica inner surface of the microplasma ion source. Both carbon and tin were detected as positively charged atomic ions, which were expelled from the microplasma ion source and directly focused by electrostatic lenses towards the quadrupole mass analyzer. Tin exhibited high selectivity to carbon (> 10(4)) and a detection limit of 3.5 pg s-1. PMID- 10457451 TI - Chemometric approach to the treatment of benzodiazepine separation and peak broadening in capillary electrophoresis. AB - A chemometric methodology was used to study capillary efficiency and the separation of ten benzodiazepines in capillary electrophoresis. The resolution between two adjacent peaks on the electropherogram was estimated and the overall quality of the separation was assessed by means of a new response function. The nature (methanol or acetonitrile) and proportion of the organic modifier both in the background electrolyte and the sample buffer and the injection time were considered. The results predicted that if the sample had a lower dielectric constant than the background electrolyte buffer then a much larger injection volume could be used. The computer optimization routine was experimentally validated and the result demonstrated that the fastest electrophoretic reparation was obtained with acetonitrile (7 min instead of 9 min with methanol). PMID- 10457452 TI - Salt effects in capillary zone electrophoresis. IV. Resolution versus time and the effect of potassium phosphate and its concentration in the high ionic strength separation of sulphonamides. AB - The effects of potassium phosphate buffer and its concentration upon the capillary zone electrophoretic separation of 23 sulphonamides and a neutral marker were examined at pH 7. The resolution between the pairs was improved with the increased concentration of the buffer from 65 mM to 174 mM. Nineteen sulphonamides, a hydrolysis product and several unidentified minor components were baseline resolved in both 101 and 138 mM phosphate buffers. In 174 mM buffer all 21 ionised sulphonamides and the other compounds were separated. A simple relationship between the resolution of analyte pairs (Rs) and the square root of the mean analysis time for the pair (square root of tapp) was derived, but few of the pairs displayed this behaviour. For the majority of pairs of compounds, Joule heating appeared to cause a maximum in the Rs versus square root of tapp relationship, while non-ideality and shifts in ionisation with increasing salt concentration appeared dominant in other cases. PMID- 10457453 TI - Sensitive, universal detection for capillary electrochromatography using condensation nucleation light scattering detection. AB - Condensation nucleation light scattering detection (CNLSD) was coupled with a pressurized capillary electrochromatography (pCEC) system using an electrospray interface. Supplementary pressure from a high-pressure pump was used to stabilize the electrospray and electrochromatography processes. Hydrodynamic injections were made with a 20 nl injection valve, and the inherent dead volume from the valve was successfully minimized, such that plate numbers in the range of 120,000 to 350,000/m were observed. Selectivity tuning using both pressure and voltage with the pressurized capillary electrochromatography system was demonstrated. Good reproducibility, comparable sensitivities for a wide range of compounds, including carbohydrates, and limits of detection down to the 50 ng/ml level, corresponding to 1-2 pg levels, were determined without the need for derivatization, demonstrating that condensation nucleation light scattering detection is a sensitive, universal detection method for pressurized capillary electrochromatography. PMID- 10457454 TI - Comparative study on the distribution of ovalbumin glycoforms by capillary electrophoresis. AB - Two commercial turkey egg ovalbumins (TEOs) with different quantities of mannose, were further purified by reversed-phase high-performance liquid chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis for either of the purified glycoproteins showed one big wide band and one close small band. Capillary electrophoresis was used for the investigation of the separation of glycoforms of both glycoproteins. The best resolution of the glycoforms was obtained, reproducibly, with 100 mM borate, 1.8 mM 1,4-diaminobutane and pH 8.6 electrophoretic buffer. At least 13 glycoform peaks could be separated for either of the two glycoproteins. Their glycoform patterns were highly similar except for the conspicuous decrease in quantity of four glycoforms in the ovalbumin containing less mannose, compared to that of the other with more mannose. Coinjection electrophoresis of the two glycoproteins indicated that almost every glycoform peak of the former exactly overlapped with its corresponding glycoform peak of the latter. These results clearly indicated that the two TEOs possessed the same glycoform patterns but differed in quantity at least four glycoforms. It was found that the glycoform patterns were remarkably different between TEO and chicken egg ovalbumin. PMID- 10457455 TI - Enantiomeric separation of D- and L-carnitine by integrating on-line derivatization with capillary zone electrophoresis. AB - A new capillary zone electrophoretic method has been developed for the enantiomeric separation and quantification of enantiomers of carnitine, D- and L carnitine were derivatized with 9-fluorenylmethyl chloroformate in a flow system, working on-line with the capillary electrophoretic equipment. The separation was performed using a selective chiral buffer containing 2,6-dimethyl-beta cyclodextrin (heptakis). Triethanolamine was used as electroosmotic modifier and the separation was carried out in a uncoated capillary. Under the optimal conditions the resolution between D- and L-carnitine was 1.2 and the limits of detection for both isomers were about 5.0 microM. The proposed method was applied to the determination of D-carnitine in excess of L-carnitine in synthetic samples, and the results demonstrated that the maximal D-:L-carnitine ratio determined was approximately 1:100. PMID- 10457456 TI - High-performance liquid chromatography coupled with fluorescence detection for the determination of trans-astringin in wine. AB - In this study a high-performance liquid chromatography (HPLC) method was developed for the determination of trans-astringin in wine using fluorescence detection. This is the first time the occurrence of trans-astringin has been reported in wine. The method allows analysis of both red and white wine samples with no prior treatment. The quantification threshold is 0.03 mg/l. Levels of trans-astringin in the French wines analyzed ranged from 0.09 mg/l to 0.29 mg/l. The reproducibility of the method was measured and the CV was less than 4.8% for both red and white wines. PMID- 10457457 TI - Quantitative structure-retention relationships of polychlorinated naphthalenes in gas chromatography. AB - A quantitative structure-retention relationship (QSRR) model has been developed for the gas chromatographic relative retention indices (RRis) of 62 polychlorinated naphthalene (PCN) congeners in a non-polar column, DB-5 (5% phenyl, methylpolysiloxane). Chemical descriptors were calculated from the molecular structures of PCNs and related to their gas chromatographic RRis by multiple linear regression analysis. The proposed model had a multiple square correlation coefficient R2 = 0.995, and standard error SE = 16.7. A QSRR reveals that the gas chromatographic retention of PCNs is associated with the number of chlorine substitutions, influenced by electronic descriptors such as heat of formation, maximum value for atomic valence, and the minimum value for electronic orbital population. PMID- 10457458 TI - Elimination of matrix effects for static headspace analysis of ethanol. AB - The intention of this work was to develop a simple and fast procedure for a determination of small amounts of ethanol in aqueous protein containing solutions based on combined headspace gas chromatography. In order to provide for short analysis time static headspace methodology was considered for this purpose. In this context the influence of the matrix composition onto the analytical results has been established and internal standardization as well as a full evaporation technique have been evaluated as promising alternatives for a compensation of matrix effects. With respect to speed of analysis, simplicity of sample handling as well as the quality of the analytical performance parameters, precision and accuracy, the full evaporation technique proved to be superior. Thus, the static equilibration of a 20 microliters sample aliquot in a conventional headspace sample vial for 5 min at 100 degrees C is sufficient to obtain equilibrium conditions for gas chromatographic analysis. The accuracy of this method was verified by robust regression analysis and exhibited excellent robustness within the required limits of sample composition ranging from 0 to 20% (w/w) protein content and up to 5 g/l salt content. PMID- 10457459 TI - Rapid, direct determination of polyphenols in tea by reversed-phase column liquid chromatography. AB - Column liquid chromatography on a C18-bonded silica column with water-methanol acetic acid as eluent was used to determine polyphenols and caffeine in tea. Without any pretreatment, catechin, epicatechin gallate, epigallocatechin gallate, epigallocatechin, epicatechin and caffeine were separated successfully within 15 min. The detection limits (S/N = 3) of polyphenols studied were 1.8-24 mg/l at a detection wavelength 270 nm. The linear range of the peak area calibration curves for the analytes were over two orders of magnitude with a correlation coefficient of 0.996-0.999. Using this method, some Chinese tea samples were analyzed with a good reproducibility (RSD are below 5%). PMID- 10457460 TI - Interpretation of chromatographic behavior of ions based on the electric double layer theory. AB - A retention model based on electrostatic theories is applied to the analysis of the ion-exchange chromatographic separation of ions. The adsorption of counterions and the ion-pair formation between ion-exchange sites and counterions are included in the model; these represent separation selectivity. A nonstoichiometric contribution, the accumulation of ions in an electrical double layer, is also involved in the model. The retention of ions is calculated by assuming these ionic properties for both eluent and solute ions. The comparison of calculated retention factors with experimental values gives insight into the ion-exchange nature of ions; e.g. a strongly adsorbed ion should have higher ion pair formation ability, and vice versa. PMID- 10457461 TI - Acoustic effects in chromatography. AB - An acoustic field effectively affects chromatographic retention in some separation modes and, thus can be a novel factor controlling retention. After being transmitted into the column, ultrasound energy is mostly converted into heat as a result of absorption by stationary and mobile phases. Thus, ultrasound brings about temperature increases. However, actual temperature increases measured by thermosensors are smaller than those calculated from chromatographic retention changes. In addition, larger ultrasound effects are observed in chromatographic modes involving ionic interactions. These results possibly imply that ultrasound directly influences ionic interactions involved in retention mechanisms. PMID- 10457462 TI - Retention modelling of electrostatic and adsorption effects of aliphatic and aromatic carboxylic acids in ion-exclusion chromatography. AB - The retention mechanism of aliphatic and aromatic carboxylic acids in ion exclusion chromatography has been investigated with consideration of simultaneous electrostatic repulsion effects and hydrophobic adsorption effects. A mathematical relationship between the retention factor of the analyte and the mobile-phase composition (sulfuric acid concentration and percentage of methanol), the type of analyte (pKa and hydrophobicity) and some physical characteristics of the stationary phase has been derived. Thirteen carboxylic acids (comprising mono- and divalent, aliphatic and aromatic acids) were chosen and used to acquire retention data on three different cation-exchange stationary phases (in which the sulfonate functional groups are bound to polystyrene divinylbenzene, polymethacrylate or silica) using 14 mobile-phase compositions of varying pH and percent methanol. These retention data were used to derive the parameters necessary to solve the retention model using non-linear regression. In this way, a quantitative measure of the effects of adsorption phenomena on analyte retention were obtained. The model was then used to optimise the separation of nine carboxylic acids. PMID- 10457463 TI - Critical comparison of retention models for the optimisation of the separation of anions in ion chromatography. II. Suppressed anion chromatography using carbonate eluents. AB - Seven theoretical retention models, namely the linear solvent strength model (using the dominant equilibrium approach and competing ion effective charge approach), the dual eluent species model, the Kuwamoto model, the extended dual eluent species model, the multiple species eluent/analyte model and the empirical end-points model, were used to describe the retention behaviour of anions in suppressed ion chromatography (IC). An extensive set of experimental retention data was gathered for 24 anions (fluoride, formate, bromate, chloride, hexanesulfonate, bromide, chlorate, nitrate, iodide, thiocyanate, perchlorate, sulfite, succinate, sulfate, tartrate, selenate, oxalate, tungstate, phthalate, molybdate, chromate, thiosulfate and phosphate) on a Dionex AS4A-SC column using carbonate eluents of varying concentration and HCO3-:CO3(2-) ratios. Statistical comparison of the predicted and experimentally obtained retention factors showed that the performance of the theoretical models improved with the complexity of the model. However the empirical model (in which a linear relationship is assumed between the logarithm of retention factor and the logarithm of eluent strength, but the slope is determined empirically) gave the most consistent performance across the widest range of anions. The empirical end-points model was also shown to be the most satisfactory model due to its low knowledge requirements and easy solution. Compared with non-suppressed IC (see Part I), the retention behaviour in suppressed IC was found to be easier to model by all retention models. PMID- 10457464 TI - Lack-of-fit testing of ion chromatographic calibration curves with inexact replicates. AB - Calibration studies involve the preparation and analysis of replicates for multiple concentrations of standards. Curves that are fitted through the data are evaluated for their adequacy of fit. A helpful test is a lack-of-fit procedure, which is performed easily by most statistical software. When coupled with Radj2, the procedure differentiates between data that are not linear and those that are simply noisy. The test requires data from exact replicates of the various standard levels involved. However, in ppt-level ion chromatography, the above condition may be impossible to meet. With the common anions (e.g., chloride, nitrite), the working standards must be prepared by mass and all liquids must be poured; transfer pipets contaminate at these concentrations. Since it is virtually impossible to pour out the desired mass exactly, final concentrations will vary slightly. Consequently, a different approach is needed for lack-of-fit testing. This paper discusses reasonable alternatives and applies them to actual data. PMID- 10457465 TI - Polyelectrolyte sorbents based on aliphatic ionenes for ion chromatography. AB - 2-4, 3-4, 2-8, 3-8, 3-6, 4-6, 6-8, 6-10-ionenes (polymers with quaternary nitrogen atoms in the main chain) served as modifiers in synthesizing polyelectrolyte sorbents for ion chromatography. The approaches to the synthesis and their stability are discussed. Cluster analysis was applied to separate aliphatic ionenes to three groups as chromatographic modifiers, namely hydrophilic, hydrophobic, and intermediate. Each group is characterized by a certain selectivity to sulfate, perchlorate, iodide and thiocyanate. The sorbents show high selectivity and efficiency up to 15,000 theoretical plates per meter. PMID- 10457466 TI - Effect of eluent composition on retention behavior of anions in ion chromatography on anion-exchangers modified with heparin. AB - Effect of eluent composition on retention behavior of inorganic anions have been investigated in ion chromatography using anion-exchangers modified with heparin. Both cation and anion of the eluent affected the retention of analyte anions and unusual retention behavior was observed on the modified stationary phase. The retention time of anions decreased with decreasing eluent concentration when sodium sulfate, magnesium sulfate and chlorides of alkali metals were used as the eluent, whereas it increased with decreasing eluent concentration when aluminum sulfate, copper sulfate and sulfuric acid were used as the eluent. The retention of nitrate increased in the order of Li+, Na+, K+, Rb+ and Cs+ when their chlorides were used as the eluent. When sodium perchlorate and chlorides of alkaline-earth metals were used as the eluent, the eluent should include heparin. Otherwise, the modifier was partially bled from the column. PMID- 10457467 TI - Capacity gradient anion chromatography with a borate complex as eluent. AB - Complex formation between borate compounds and vicinal diols is well recognized. Generally, in a chemically bonded anion-exchange resin, many hydroxyl groups are introduced on the surface of the resin in order to make the resin hydrophilic. The borate as an eluting reagent also reacts to these hydroxyl groups, and this complex formation decreases the apparent ion-exchange capacity of the column by being dissociated to the anion depending on the eluent pH. In the present work a method is described for the simultaneous determination of anions based on the capacity gradient for suppressed ion chromatography. A Tosoh IC-Anion-PW column and dihydroxyphenylborane-mannitol eluent system were used. To maintain baseline stability, it was helpful to keep the borate concentration constant during a gradient of 16 to 0 mM mannitol as a modifier to prevent the complex formation with the hydroxyl on the resin. The chemical composition of the eluents and gradient profiles are discussed and the application to the analysis of the condensed phosphates with widely varying retention times as food additives in a cheese sample is presented. PMID- 10457468 TI - Simultaneous determination of anions and cations by ion-exclusion chromatography cation-exchange chromatography with tartaric acid/18-crown-6 as eluent. AB - Ion-exclusion chromatography-cation-exchange chromatography was developed for the simultaneous separation of common inorganic anions and cations (Cl-, NO3- and SO4(2-); Na+, NH4+, K+, Mg2+ and Ca2+) on a weakly acidic cation-exchange column by elution with weak acid. Generally, the resolution among these monovalent cations was only moderate, thereby hindering the determination of these analytes in natural-water samples. Therefore, 18-crown-6 was added to the eluent to improve the resolution. A good separation of these anions and cations on a weakly acidic cation-exchange column was achieved in 30 min by elution with 5 mM tartaric acid/6 mM 18-crown-6/methanol-water (7.5:92.5). The ion-exclusion chromatography-cation-exchange chromatography method developed here was successfully applied to the separation of major anions and cations in an environmental water sample. PMID- 10457469 TI - Enhancement of conductometric detection of weak acids in ion chromatography. AB - Anions of weak acids can represent a problem when determined via chemically suppressed ion chromatography as the acids can be weakly ionised, giving low conductivity and hence low sensitivity. Previous work showed that converting some weak acids back to their sodium salts, using a second micromembrane suppressor, greatly enhanced conductivity and thus sensitivity. This paper will discuss further work in optimising the conversion for boric acid by using sodium salts of EDTA and the mechanisms involved. PMID- 10457470 TI - Combination of suppressed and non-suppressed ion chromatography with atmospheric pressure ionization mass spectrometry for the determination of anions. AB - Non-suppressed and suppressed ion chromatography in combination with atmospheric pressure ionization mass spectrometry are compared with special respect to sensitivity for the analysis of low-molecular-mass anions. Iodate, bromate, bromide, sulfate, thiosulfate and bromide could be separated by non-suppressed ion chromatography using a low-capacity anion-exchange column and ammonium citrate as mobile phase. Absolute detection limits between 0.4 and 0.7 ng could be achieved; employing a column requiring a flow-rate of 1 ml/min for optimum performance, splitting was necessary so that only 120 microliters/min entered the interface of the mass spectrometer resulting in detection limits between 0.03 and 0.06 mg/l. The same stationary phase (packed into a narrow-bore column which allowed operation without splitting) was suitable for the separation of oxyhalides in the suppressed mode with detection limits of 0.5 microgram/l (50 pg) with sodium carbonate as eluent. The method was applied to the analysis of drinking water for oxyhalides. The sample pretreatment for the removal of matrix anions (sulfate, chloride and hydrogencarbonate) is described. PMID- 10457471 TI - Low-level calibration study for a new ion chromatographic column to determine borate in deionized water. AB - In the semiconductor industry, there is interest in determining borate at sub-ppb levels in ultrapure water, since borate is an early breakthrough ion from ion exchange resin beds. Although dissolved silica is the most common species currently used to monitor the breakdown of the deionization systems, it is thought that borate probably breaks through earlier than silicate. To be of use as an early-warning indicator, borate must be determined at ppt levels. This paper discusses benchtop results with several new column products designed to deliver low-ppt detection limits for boron as borate. The system uses a prototype borate-specific concentrator column that is coupled to an ion-exclusion separator and suppressed-conductivity detection. The acidic eluent, containing mannitol, quantitatively elutes the borate from the concentrator. The analytical separation is performed using a specially designed ion-exclusion column. Data presented are from two multilevel calibration studies. Included is a discussion of detection limit calculations and recommended formats for reporting results. PMID- 10457472 TI - Analysis of 500-ng/l levels of bromate in drinking water by direct-injection suppressed ion chromatography coupled with a single, pneumatically delivered post column reagent. AB - In July 1997, the US Environmental Protection Agency (EPA) began sampling and analyzing drinking water matrices from US municipalities serving populations greater than 100,000 for low-level bromate (> 0.20 microgram/l) in support of the Information Collection Rule (ICR) using the selective anion concentration (SAC) method. In September 1997, EPA published Method 300.1 which lowered the Method 300.0 bromate method detection limit (MDL) from 20.0 to 1.4 micrograms/l. This paper describes the research conducted at the EPA's Technical Support Center laboratory investigating a single post-column reagent, o-dianisidine (ODA), which has been successfully coupled to EPA Method 300.1 to extend the MDL for bromate. Initial studies indicate that this method offers a MDL which approaches the EPA's SAC method with the added benefit of increased specificity, shortened analysis time and reduced sample preparation. The method provides excellent ruggedness and acceptable precision and accuracy with a bromate MDL in reagent water of 0.1 microgram/l, and a method reporting limit of 0.50 microgram/l. PMID- 10457473 TI - Determination of trace level perchlorate in drinking water and ground water by ion chromatography. AB - Ammonium perchlorate, a key ingredient in solid rocket propellants, has recently been found in ground and surface waters in the USA in a number of states, including California, Nevada, Utah, and West Virginia. Perchlorate poses a health risk and preliminary data from the US Environmental Protection Agency reports that exposure to less than 4-18 micrograms/l provides adequate human health protection. An ion chromatographic method was developed for the determination of low microgram/l levels of perchlorate in drinking and ground waters based on a Dionex IonPac AS11 column, a 100 mM hydroxide eluent, large loop (1000 microliters) injection, and suppressed conductivity detection. The method is free of interferences from common anions, linear in the range of 2.5-100 micrograms/l, and quantitative recoveries were obtained for low microgram/l levels of perchlorate in spiked drinking and ground water samples. The method detection limit of 0.3 microgram/l permits quantification of perchlorate below the levels which ensure adequate health protection. A new polarizable anion analysis column, the IonPac AS16, and its potential applicability for this analysis is also discussed. PMID- 10457474 TI - Determination of total chromium in phosphate rocks by ion chromatography. AB - Chromium(VI) is one of seven elements which is classified in the fertilizer industry as being harmful to plants and biological systems. Phosphate rocks represent the raw material for complex fertilizer production in the world. This paper investigates for the first time the determination of total chromium in phosphate rocks by ion chromatography. The developed analytical method involves the digestion of phosphate rocks with nitric acid followed by sample treatment of the resulting solution. The digestion solution obtained was treated with an oxidising agent (potassium peroxosulphate) to convert all chromium to the hexavalent state. The analytical method developed utilizes anion-exchange ion chromatography to achieve the separation and spectrophotometric post-column reaction for detection with diphenylcarbazide. The relative standard of deviation from analytical data comparison of six different phosphate rocks with atomic absorption spectrometry and inductively coupled plasma atomic emission spectrometry techniques, and cross-analysis data against an internationally certified phosphate rock standard were between 0.58 and 1.45%. Calibration curve between 0.2 and 0.9 microgram/ml was excellent, and the method has a detection limit for Cr(VI) of 0.05 ng. The developed method offers a fast, a reliable and an alternative procedure for the determination of total chromium in phosphate rock deposits by ion chromatography. PMID- 10457475 TI - Anion-exchange separation and determination of bisphosphonates and related analytes by post-column indirect fluorescence detection. AB - Bisphosphonic acids and their salts can be detected after their liquid chromatographic separation by post-column indirect fluorescence detection (IFD). After separation the analyte is combined with the highly fluorescent Al(3+)-morin (2',3,4',5,7-pentahydroxyflavone) solution and fluorescence decreases because of the formation of the nonfluorescent Al(3+)-bisphosphonate complex. The decrease in fluorescence is proportional to the amount of bisphosphonate present. Separation of the multivalent anionic bisphosphonate analytes from other anions and sample matrix is achieved on a strong base anion-exchange column with a strong, basic eluent. The post-column reaction variables, which influence IFD, are identified and optimized for the detection of the bisphosphonates after separation on the anion exchanger. The method is selective, since only a few anions will undergo a reaction with the Al(3+)-morin solution, and sensitive, detection limit for difluoromethylene bisphosphonate, F2MDP, is 4 ng for S/N = 3. The separation-IFD method can be applied to the determination of bisphosphonates, such as F2MDP, ethane-1-hydroxy-1,1-bisphosphonic acid, dichloromethylene bisphosphonic acid, 4-amino-1-hydroxybutane-1,1-bisphosphonic acid, in biological samples. The separation-IFD method is also applicable to the detection of inositol phosphate anions. PMID- 10457476 TI - Ion chromatography of nitrite at the ppb level with photometric measurement of iodine formed by post-column reaction of nitrite with iodide. AB - The difficulty in ion-chromatographic determination of nitrite in aqueous solutions containing a high concentration of chloride arises mainly from incomplete resolution of the peaks for these anions on the separation column whose efficiency is not high. A photometric measurement of iodine formed by a reaction of nitrite with iodide has been found to make it possible to determine, chromatographically, trace amounts of nitrite without any interference from chloride; chloride does not oxidize iodide to produce iodine. The proposed method was based on the separation of nitrite from matrix anions on a silica-based anion exchange column with a 1.5.10(-3) M phthalate eluent (pH 5.0), followed by photometric measurement of the iodine (as triiodide) formed via a post-column reaction of the separated nitrite with iodide. The optimal conditions for the post-column reaction were established by varying the concentrations of iodide, copper(II) and nitric acid in a post-column-reaction solution and the length of a reaction tube. A calibration graph for nitrite, plotted as peak heights versus concentrations, was linear up to 1.50.10(-5) M (690 ppb). The detection limit, defined at S/N = 3, was 1.00.10(-7) M (4.60 ppb) nitrite. The presence of chloride ions up to 0.01 M did not give any interference to the determination of nitrite. This method was successfully applied to the determination of nitrite in lake water, river water, sewage works water and snow samples without any pretreatment. PMID- 10457477 TI - Electrostatic ion chromatography of polarizable anions in saline waters with N-[2 [acetyl(3-sulfopropyl)amino]ethyl]-N,N-dimethyldodecanaminium hydroxide (ammonium sulfobetaine-1) as the stationary phase and a dilute electrolytic solution as the mobile phase. AB - A new type of zwitterionic surfactant, N-{2-[acetyl(3-sulfopropyl)amino]ethyl} N,N-dimethyldodecanaminium hydroxide (ammonium sulfobetaine-1), with a greater distance between the two charged groups, was used as the stationary phase for electrostatic ion chromatography (EIC) of polarizable anions (e.g., thiocyanate, iodide and nitrate) in saline water samples. The targeted species (polarizable anions) were baseline separated using this type of zwitterionic surfactant as the stationary phase, but the highly polarizable species (iodide and thiocyanate) were eluted faster (compared with the results obtained using N-dodecyl-N,N dimethyl-3-ammonio-1-propanesulfonate, C12N3S, with a shorter distance between the two charged groups, as the stationary phase). In other words, the extent of binding of the highly polarizable anion (iodide and thiocyanate) was found to be smaller when using ammonium sulfobetaine-1 as the stationary phase. This provides a rapid but effective method for the analysis of highly polarizable anions in saline water samples. The results for the successful detection of iodide in seawater demonstrates the usefulness of this new type of zwitterionic surfactants for EIC. PMID- 10457478 TI - Determination of trace anions in concentrated weak acids by ion chromatography. AB - Ion-exclusion chromatography (ICE) followed by ion chromatography (IC) was used for the determination of trace anionic contaminants in concentrated weak acids. The ICE separation was used as a pretreatment step to isolate the contaminant anions of strong acid from the excess of matrix ions. Then a fraction containing the analyte ions was separated using IC with suppressed conductivity detection. Microbore-ion-exchange columns were chosen to address the increased purity requirements for use of these concentrated acids in semiconductor applications. The chromatographic conditions were optimized for determining trace chloride, sulfate, phosphate, and nitrate in concentrated 24.5% (v/v) hydrofluoric acid; trace chloride, sulfate, and nitrate in concentrated 85% (w/w) phosphoric acid and trace chloride and sulfate in concentrated 0.7% (v/v) glycolic acid. Method detection limits for the anions of interest were below 100 micrograms/l. PMID- 10457480 TI - Separation of carboxylic acids on a weakly acidic cation-exchange resin by ion exclusion chromatography. AB - The separation of various carboxylic acids was performed on a polymethacrylate based weakly acidic cation-exchange resin (TSKgel OApak-A) using ion-exclusion chromatography under the acidic elution conditions. When a diluted sulfuric acid solution was used as the eluent, highly sensitive conductimetric detection of carboxylic acids was achieved without increasing the background conductance of the eluent. This method was more sensitive than using benzoic acid eluent and enabled a good resolution of dicarboxylic as well as monocarboxylic acids. The addition of 5-20% methanol to the eluent considerably reduced the retention times of carboxylic acids with hydrophobic nature. PMID- 10457479 TI - Ion-exclusion chromatography of carboxylic acids on silica gel modified with aluminium. AB - The modification of silica gel with aluminium by a coating method was effective for the preparation of a silica-based stationary phase, which acted as a cation exchanger under strongly acidic conditions. In order to expand the utility of the laboratory-made aluminium-adsorbing silica gel it was applied as a stationary phase to the ion-exclusion chromatography of various carboxylic acids. Good separations for both aliphatic carboxylic acids and benzenecarboxylic acids with a hydrophobic nature under acidic eluent conditions were achieved in 25 min. PMID- 10457481 TI - Preconcentration and separation of haloacetic acids by ion chromatography. AB - A comparative study was made of the chromatographic behaviour of five haloacetic acids (mono-, dibromoacetic and mono-, di-, trichloroacetic acids). The techniques investigated included reversed-phase ion interaction chromatography with UV detection, suppressed and non-suppressed anion-exchange chromatography. The systems are discussed studying the retention as a function of the mobile phase parameters and the stationary phases used (LiChrospher 100 RP-18, IonPac AS9, AS10 and AS11). A preconcentration step, performed on different substrates, namely LiChrolut-EN and activated vegetal carbon, has been optimized in order to reduce the method detection limits. Results obtained for drinking water samples are shown. PMID- 10457482 TI - Liquid chromatography: a tool for the analysis of metal species. AB - An overview is presented of classic and more recent applications of liquid chromatography for the analysis of metal species. The different approaches involving ion-exchange, ion-pair, and chelation separation mechanisms are discussed as well as the new philosophy of simply removing interferents before specific detections of metal ions (alkali and alakaline earths, rare earths, heavy and transition metals). New more selective materials enabling difficult separations and studies on multimodal or hyphenated techniques for metal speciation (e.g. arsenic and chromium) are considered. PMID- 10457484 TI - Determination of sodium and ammonium ions in disproportionate concentration ratios by ion chromatography. AB - In ion chromatography, samples of very different ammonium-to-sodium concentration ratios are difficult to quantify since these two cations have similar selectivities for stationary phases containing commonly used sulfonate or carboxylate cation-exchange functional groups. The IonPac CS15 cation-exchange column, with carboxylate and phosphonate functional groups as well as a crown ether group, was developed to address this limitation. Selectivity for the common inorganic cations on this column is different from that of conventional cation exchange columns in that the separation between sodium and ammonium ions has been greatly increased, allowing for determinations of low levels of one in the presence of high levels of the other with an isocratic eluent. For larger than 4000:1 sodium-to-ammonium concentration ratios, an eluent step change or gradient elution is needed. For moderate ratios, combinations of this column with a carboxylate column, containing no crown ether group, can be used at room temperature with an isocratic eluent containing no organic solvent. PMID- 10457483 TI - Simultaneous separation of common mono- and divalent cations on silica gel modified with aluminium by non-suppressed ion chromatography with conductimetric detection and nitric acid-15-crown-5 as eluent. AB - The modification of silica gel with aluminium by a coating method was very effective for the preparation of silica-based stationary phases which acted as a cation exchanger under strongly acidic conditions. However, the separation of common mono- and divalent cations (Li+, Na+, NH4+, K+, Mg2+ and Ca2+) on an aluminium-adsorbing silica (Al-Silica) column was moderate by a conductimetric detection ion chromatography (IC) with strongly acidic eluents. Then, the addition of various crown ethers (12-crown-4, 15-crown-5 and 18-crown-6) in acidic eluent was carried out. As a result, it was found that 15-crown-5 was most effective for the improvement of peak resolution. Excellent separation of these cations was achieved in 20 min by elution with 2 mM nitric acid-2 mM 15-crown-5. The proposed IC was successfully applied to the determination of major cations in various natural waters. PMID- 10457485 TI - Separation and detection of common mono- and divalent cations by ion chromatography with an ODS column and conductivity/UV detection. AB - The retention and detection behavior of common mono- and divalent cations (M+, alkali metal (Li+, Na+, K+, Rb+, Cs+) and ammonium ions (NH4+); M2+, alkaline earth metal ions (Mg2+, Ca2+, Sr2+, Ba2+) was examined using an ODS column (150 x 4.6 mm I.D.) and conductivity (CD)/UV detection. The results obtained were as follows: (1) for M+, the mobile phase, 0.1 mM sodium dodecyl sulphate (SDS) + 10 mM HNO3 and indirect CD detection were effective. (2) Addition of Ce(III) in the mobile phase accelerated the elution of both M+ and M2+. The separation of above 10 cations on an ODS column was achieved for the first time without any coelution of cations and disturbance by system peak. Addition of higher SDS resulted in good separation of M+ and M2+ with longer retention times. CD detection was possible for M+ and M2+ and UV detection for M2+. (3) For M2+, the mobile phase, 0.8 mM Ce(III) + 0.1 mM SDS + 1 mM HNO3 and indirect UV detection were effective. The IC methods were applied to real samples. PMID- 10457486 TI - Determination of niobium(V) and tantalum(V) as 4-(2-pyridylazo)resorcinol-citrate ternary complexes in geological materials by ion-interaction reversed-phase high performance liquid chromatography. AB - A method for the simultaneous separation and determination of Nb(V) and Ta(V) as ternary complexes formed with 4-(2-pyridylazo)resorcinol (PAR) and citrate was developed using ion-interaction reversed-phase high-performance liquid chromatography on a C18 column. Method parameters, such as pre-column complex formation conditions and composition of the complexes were investigated using spectrophotometry and HPLC. Under the optimum conditions, the Nb(V) and Ta(V) complexes were eluted within 12 min with a mobile phase of methanol-water (32:68, v/v) containing 5 mM acetate, 5 mM TBABr and 5 mM citrate buffer at pH 6.5, with detection at 540 nm. A typical separation efficiency was 33,000 and 20,000 theoretical plates per metre for Nb(V) and Ta(V), respectively. The relative standard deviation of retention times for the Nb(V) and Ta(V) complexes were 0.16% and 0.17% and for peak areas were 0.28% and 1.36%, respectively. The detection limits (signal-to-noise ratio = 3) for Nb(V) and Ta(V) were 0.4 ppb and 1.4 ppb, respectively. Results obtained for standard reference rock samples agreed well with certified values and results obtained by inductively coupled plasma MS. PMID- 10457487 TI - Separation selectivity of some ethylenediaminetetraacetic acid and cyclohexane 1,2-diaminetetraacetic acid complexes in column and ion electrokinetic chromatography. AB - The complexes of Mn2+, Cd2+, Fe3+, Pb2+, Ni2+, Co2+, Zn2+ and Cu2+ with EDTA and cyclohexane-1,2-diaminetetraacetic acid (CDTA) were separated and detected in column and ion electrokinetic chromatography with suppressed conductivity and direct UV detection, respectively. In column ion chromatography (IC) these complexes were separated on an IonPac AS4A anion-exchange column (Dionex, USA). Parameters of carrier electrolyte, which were examined in the ion electrokinetic chromatography (IEKC) mode, include polymer and sulfate concentrations. In IEKC separation selectivity of complexes with poly(diallyldimethylammonium) cation as modifier is similar as for an IonPac AS4A column both for EDTA and CDTA chelates. It was shown that the ion-exchange capacity of the electrokinetic system is more than 100-times lower than the capacity of the IC column for the same peak resolution. In comparison with column main advantages of electrokinetic version are high separation efficiency (220,000-390,000 theoretical plates) and the absence of the analyte interaction with the sorbent matrix. PMID- 10457488 TI - Determination of aspartame by ion chromatography with electrochemical integrated amperometric detection. AB - In this paper, the separation and determination of the sweetener aspartame by ion chromatography coupled with electrochemical amperometric detection is reported. Sodium saccharin, acesulfame-K and aspartame were separated using 27.5 mmol/l NaOH isocratic elution on a Dionex IonPac AS4A-SC separation column. Aspartame can be determined by integrated amperometric detection without interference from the other two sweeteners. The method can be applied to the determination of aspartame in powered tabletop, fruit juice and carbonated beverage samples, and the results obtained by integrated amperometry were in agreement with those obtained using a UV detection method. A method for determining analytes with an NH2 group by ion chromatography with integrated amperometry was developed. PMID- 10457489 TI - Applications of ion chromatography in the semiconductor industry. II. Determination of basic airborne contaminants in a cleanroom. AB - Since the geometry of semiconductors and integrated circuits has been shrunk to well below sub-micron dimensions, there is a great demand for precise and reliable analytical techniques to measure and monitor the contaminants in all areas related to the fabrication process. Special concerns about the air cleanliness in a Fab lead to the necessity for developing analytical techniques to perform this task. In this research, basic airborne contaminants in a cleanroom were adsorbed onto a collection tube, subsequently extracted with deionized water and analyzed by ion chromatography. Such a method is capable of simultaneously measuring the concentrations of ammonia and cations (e.g., sodium, potassium, calcium, magnesium, etc.) present in the cleanroom air samples. The optimal sample preparation method was determined and the analyte concentrations at various locations in the cleanroom were measured. The results showed significant variations from one location to another. The long-term fluctuations in the contaminant levels were also significant. Data obtained using this method compared well with data from inductively coupled plasma analysis of the same materials. PMID- 10457490 TI - Determination of amines used in the oil and gas industry (upstream section) by ion chromatography. AB - During production and purification of crude oil and natural gas several different amines are used as chemicals or operating materials, e.g. film forming long chain amines as corrosion inhibitors, steam volatile amines for pH correction and corrosion protection, alkanolamines as absorbents in sour gas treatment plants, etc. For analytical checks, e.g. determination of corrosion inhibitor concentration in produced media, classical chemical methods are used predominantly, because most of them can be performed in small field laboratories. Some amines, especially the small molecular aliphatic and heterocyclic amines can also be determined by ion chromatography. In our laboratory two types of separation columns (IonPac CS10 and CS12A) were available for ion chromatographic separation. The analysis of the amines in low-salt-containing water, soft water or steam condensate can be performed without problems. The presence of alkali and/or alkaline earth ions in the sample can lead to coelution with these ions, to poor peak resolution or enhanced analysis times, depending on the chromatographic conditions. This work shows some examples of ion chromatography applications for the determination of low-molecular-mass ethanolamines, morpholine and piperazine and discusses the possible interferences and troubles caused by alkali and alkaline earth ions in the matrix. PMID- 10457491 TI - Determination of glyphosate by ion chromatography. AB - An ion chromatography system for the determination of glyphosate was described. Ion chromatograph was carried out by suppressed conductivity detection (DX-100). The eluent contained 9 mmol l-1 Na2CO3 and 4 mmol l-1 NaOH. The detection limit was 0.042 microgram ml-1 (S/N = 3). The relative standard deviation was 1.99% and the correlation coefficient of the calibration curve for area was 0.9995. The linear range was 0.042-100 micrograms ml-1. Common inorganic ion and organic acids did not interfere. The recovery was 96.4-103.2%. The method was simple, rapid, reliable and inexpensive. PMID- 10457492 TI - Analysis of wastewater for anionic and cationic nutrients by ion chromatography in a single run with sequential flow injection analysis. AB - To prevent nutrient enrichment and, hence the undesirable ecological impacts, the nutrients monitored in wastewater samples include two anionic species, i.e., nitrate and orthophosphate, and a cationic species, ammonium. Ion chromatography (IC) is one of the popularly used techniques for determinations of nitrate and phosphate in these samples, whereas determination of ammonium in wastewater samples is typically done using manual or automated wet chemistry, e.g., flow injection analysis (FIA). We have developed a sequential IC-FIA method, using Lachat's QC8000 IC system, which allows determinations of nitrate, phosphate and ammonia in a single injection. In this system, a QuickChem Small Suppressor cartridge is regenerated in between the samples. A sample is injected while leaving the suppressor off-line. Ammonium, a cation, elutes in the void volume of an anion-exchange column. The unsuppressed column effluent, exiting the conductivity flow cell, up to this point is used for FIA determination of ammonia. When ammonia exits the conductivity flow cell, a fully regenerated suppressor is brought in-line for conductometric detection of the anions. Analog data are simultaneously acquired from colorimetric and conductometric detectors, for the cationic and anionic nutrients, respectively. The method is accurate with spike recoveries in wastewater samples ranging from 91% for nitrate to 114% for chloride. It is precise with RSD values, for replicate analyses (n = 7) of a mid range standard, ranging from 0.4% for phosphate to 1% for nitrate. PMID- 10457493 TI - Acid-rain monitoring in east Asia with a portable-type ion-exclusion-cation exchange chromatographic analyzer. AB - A monitoring system consisting of a portable-type conductimetric ion-exclusion cation-exchange chromatographic (CEC) analyzer and a meteorological satellite data analyzer has been investigated for the evaluation of the effects of acid precipitation on natural and urban environments in East Asia. The portable ion exclusion-CEC analyzer uses a polymethacrylate-based weakly acidic cation exchange resin column in the H(+)-form and a weak-acid eluent (tartaric acid methanol-water) and is applied for the simultaneous determination of anions (SO4(2)-, NO3-, and Cl-) and cations (Na+, NH4+, K+, Mg2+, and Ca2+) in precipitation transported from mainland China to central Japan, as mapped by the meteorological satellite data analyzer. Linear calibration graphs of peak area versus concentration for anions and cations were observed in the concentration range 0-1.0 mM for the anions and 0-0.5 mM for the cations. Detection limits at a signal-to-noise ratio of 3 were in the range 5.18-12.1 ppb for the anions and 6.58-16.5 ppb for the cations. The practical utility of this monitoring system is presented. PMID- 10457494 TI - Factors affecting selectivity of inorganic anions in capillary electrophoresis. AB - Capillary zone electrophoretic separations of inorganic anions are largely governed by the intrinsic (infinite dilution) mobility of the anion. This in turn is a function of the hydrodynamic friction caused by the size of the ion and the dielectric friction caused by the charge density of the anion re-orienting the surrounding solvent. The influence of these factors on the mobility of anions is examined in both water and nonaqueous solvents. The influence of other experimental parameters, such as ionic strength, ion association, electroosmotic flow modifier concentration, and the addition of complexing agents such as polymeric cations, cyclodextrins, crown ethers and cryptands are also reviewed. From this discussion, some rules of thumb as to when different approaches will be most effective are drawn. PMID- 10457495 TI - Sample stacking of fast-moving anions in capillary zone electrophoresis with pH suppressed electroosmotic flow. AB - On-line sample concentration of fast moving inorganic anions by large volume sample stacking (LVSS) and field enhanced sample injection (FESI) with a water plug under acidic conditions is presented. Detection sensitivity enhancements were around 100 and 1000-fold for LVSS and FESI, respectively. However, reproducibility and linearity of response in the LVSS approach is superior compared to the FESI approach. PMID- 10457496 TI - Artificial neural networks for computer-aided modelling and optimisation in micellar electrokinetic chromatography. AB - The separation process in capillary micellar electrochromatography (MEKC) can be modelled using artificial neural networks (ANNs) and optimisation of MEKC methods can be facilitated by combining ANNs with experimental design. ANNs have shown attractive possibilities for non-linear modelling of response surfaces in MEKC and it was demonstrated that by combining ANN modelling with experimental design, the number of experiments necessary to search and find optimal separation conditions can be reduced significantly. A new general approach for computer aided optimisation in MEKC has been proposed which, because of its general validity, can also be applied in other separation techniques. PMID- 10457497 TI - Optimization of conductivity detection of low-molecular-mass anions in capillary zone electrophoresis. AB - The optimization of background electrolyte compositions for capillary zone electrophoresis in combination with conductivity detection focusing on maximal detector response is discussed. A theoretical approach pointing out the influence of the electrolyte co- and counter-ion mobilities on the detector signal has been developed. Using this model, running buffer compositions providing optimum S/N ratios for the selected analytes could be calculated. The results derived from these examinations have been verified by experimental investigations, namely the determination of inorganic and organic anionic solutes. PMID- 10457498 TI - Development of an analytical method for beryllium in airborne dust by micellar electrokinetic chromatography. AB - The separation of diacetylacetonato-beryllium [Be(acac)2] from acetylacetone was achieved with micellar electrokinetic chromatography (MEKC) followed by subsequent beryllium analysis carried out using capillary electrophoresis. Analysis using a separation and absorption detector, with a 50 microns capillary cell, could detect approximately 1000 ppb of beryllium in the final sample. Be(acac)2 is a promising reagent for absorption spectrophotometry, because its molar absorption coefficient, (32,000 l mol-1 cm-1) is very large. As the complex and acetylacetone have a similar absorption wavelength, the excess free ligand must removed from the sample to be measured. Acetylacetone is a weak acid with keto-enol tautomerism in aqueous solution. Acetylacetone and neutral Be(acac)2 should be separated with capillary zone electrophoresis (CZE) using a neutral or basic buffer solution as the mobile phase. Although the pH and temperature of the mobile phase were optimized, separation with CZE was interfered with by a portion of acetylacetone. This interfering portion seems to be a neutral keto-form with the same migration time as Be(acac)2. As a neutral species separation method, MEKC with sodium dodecyl sulfate was tried and the separation was completed. The optimum pH value and buffer temperature are pH 7.8 and 15 degrees C, respectively. PMID- 10457499 TI - Determination of melatonin in biological samples by capillary electrophoresis. AB - The determination of melatonin (MLT) in physiological samples was investigated using capillary electrophoresis (CE). Mouse blood was collected in tubes containing EDTA, centrifuged at 1500 g for 20 min at 4 degrees C, and stored at 20 degrees C. Plasma samples were extracted with dichloroethane, centrifuged and the aqueous phase was discarded. Then the organic phase was evaporated to dryness. The residue was dissolved in deionized water and filtered with a microfilter (0.22 micron). Separations were carried out using a CE system equipped with a fused silica capillary [80 cm (effective length 52 cm) x 75 microns I.D.] and an ultraviolet-visible detector (200 nm), and programmed to provide 25 mM 2-(N-morpholino)ethanesulfonic acid (pH 5.7). Injection was performed hydrostatically by elevating the sample by 10 cm at the cathodic side of the capillary. The calibration curve, reproducibility, recovery and limit of detection were examined, and validation of the method was performed. The result showed that MLT in blood could be easily determined with the new method. PMID- 10457500 TI - Chemical conditions inside occluded regions on corroding aircraft aluminum alloys. AB - Corrosion of aluminum alloy structures costs the US Air Force in the order of US$1 x 10(9) annually. Corrosion develops in areas of overlap such as aircraft lap-splice joints and under protective organic coatings. Capillary electrophoresis (CE) has been used to determine the local chemistries at these corrosion sites of solutions that were extracted using a microsampling system. Analysis of the local solution within lap-splice joints from aircraft has been performed in two ways: rehydration of corrosion products and direct microsampling. The solutions collected were analyzed with CE to quantitatively determine the species present during corrosion. The most common ions detected were Cl-, NO2-, NO3-, HCO3-, K+, Al3+, Ca2+, Na+ and Mg2+. Studies of the solution chemistry under local coating defects are required to understand coating failure and develop more durable coatings. A microsampling system and micro pH sensor were developed to extract solution from and measure pH in defects with diameters as small as 170 microns. Actively corroding defects contained high concentrations of Cl-, Al3+, Mg2+, Mn2+ and Cu2+ whereas only trace levels of Mg2+ were found in repassivated defects. The effects of these species on initiation and propagation of corrosion are discussed. PMID- 10457501 TI - Using capillary electrophoresis to study the chemical conditions within cracks in aluminum alloys. AB - The environment-assisted cracking (EAC) susceptibility of some aluminum alloys used for airplane structural components currently limits their use in the peak strength condition. Understanding the mechanism of EAC will facilitate the development of crack-resistant alloys with optimum mechanical properties. One component towards understanding the fundamental processes responsible for EAC is a comprehensive knowledge of the chemical conditions within cracks. The present work uses capillary electrophoresis (CE) to quantify the crack chemistry in order to provide insight into the nature of the mechanism controlling cracking. The highly restricted geometry of cracks in metals means that a crack typically contains less than 10 microliters of solution. The high mass sensitivity combined with the inherently robust nature of CE makes it an ideal analytical technique for this application. Complicating factors in the accurate determination of the crack environment include high levels of sodium present from the test solution. Low sample volume and analyte matrix complexity necessitated the development of specific sampling, extraction and analysis methods. Analysis of the crack solutions in EAC-susceptible material revealed high levels of Al3+, Mg2+, Zn2+, and Cl- near the crack tip. Cations arise from the anodic dissolution of the alloy, whereas chloride ingress from the external environment occurs to maintain solution electroneutrality within the crack. In contrast, EAC-resistant material exhibited significantly lower concentrations of dissolution products. PMID- 10457502 TI - Laboring for longevity. An annotated poem. PMID- 10457503 TI - Urine drug screening: can counterfeit urine samples pass inspection? PMID- 10457504 TI - Controlling workers' compensation medical care use and costs through utilization management. AB - Little is known about the performance of utilization management (UM) programs, which are now widely used within the workers' compensation system to contain medical costs and improve quality. UM programs focus largely on hospital care and rely on preadmission and concurrent reviews to authorize hospital admissions and continued stays. We obtained data from a large UM program representing a national sample of 9319 workers' compensation patients whose medical care was reviewed between 1991 and 1993. We analyzed these data to determine the denial rate for hospital admission and outpatient surgery and the frequency of length-of-stay restrictions among hospitalized patients. The denial rate was approximately 2% to 3% overall, but many of the denials were later reversed. On average, the UM program reduced the length of stay by 1.9 days relative to the number of days of care requested. The estimated gross cost savings resulting from reduced hospitalization time and decreased outpatient care was approximately $5 million. UM programs may offer a viable approach to cost containment within the workers' compensation system. Their value as a tool to improve the quality of care for workers' compensation patients remains to be demonstrated. PMID- 10457505 TI - Occupational exposure to antineoplastic agents: self-reported miscarriages and stillbirths among nurses and pharmacists. AB - Insult to the germ cells of an ovum or sperm prior to pregnancy as well as exposures to a fetus during pregnancy can affect the outcome of a pregnancy. Antineoplastic agents are mutagenic and teratogenic, so the potential effects of exposure on reproduction are of concern to the workers who handle them. This study investigates pregnancy loss associated with occupational exposures to antineoplastic drugs by comparing rates of spontaneous abortion and stillbirths for pregnancies without antineoplastic exposure and exposed pregnancies in which the pregnant woman or the father handled antineoplastic agents either before or during the pregnancy. A total of 7094 pregnancies of 2976 pharmacy and nursing staff were examined. After age during pregnancy, prior gravidity, maternal smoking during the pregnancy, and occurrence of a spontaneous abortion or stillbirth in a prior pregnancy were controlled for, exposure of the mother to or the handling of antineoplastic agents during the pregnancy was associated with a significantly increased risk of spontaneous abortion (odds ratio = 1.5; 95% confidence interval, 1.2 to 1.8) and combined risk of spontaneous abortion and stillbirth (odds ratio = 1.4; 95% confidence interval, 1.2 to 1.7) but not stillbirth alone. Among the wives of exposed men, too few stillbirths occurred to allow analysis. However, for spontaneous abortion and any loss, the patterns of increased risk were similar to those seen for women, although the odds ratios were not statistically significant. PMID- 10457506 TI - Ethical issues in the use of genetic markers in occupational epidemiologic research. AB - This review was conducted to characterize the nature of contemporary occupational epidemiologic research involving genetic markers, consider how genetic information is unique with regard to its social applications, and examine some of the ethical dilemmas that may arise over the course of studies. We have reviewed the literature and the lessons from our experience in conducting occupational epidemiologic research involving genetic markers. This review describes how occupational epidemiologic studies differ from other epidemiologic studies on issues of participation, confidentiality, and the history of including genetic markers. Of primary concern in occupational studies are genes that have multiple alleles and are sometimes referred to as "metabolic polymorphisms." They generally do not confer risk on their own but rather only in combination with a specific exposure. There is a need for a clear policy and guidelines for the conduct of occupational epidemiologic studies using genetic material. This policy should address all of the steps in study design, implementation, interpretation, and communication of results. PMID- 10457507 TI - Workplace-violence investigations by the California Division of Occupational Safety and Health, 1993-1996. AB - In response to the growing recognition of violence in the workplace, government agencies who oversee workplace safety are beginning to incorporate security issues into comprehensive safety plans. The California Division of Occupational Safety and Health began conducting inspections for violent events in 1993, and information from these security inspections was collected from January 1993 through January 1997. The inspections were examined by initiating source, industry, type of event, and citations issued and compared with distributions of known victimizations. The factors predicting whether a citation was issued were determined through logistic models. Over 200 inspections were conducted in 11 industries, with retail and health care establishments inspected most frequently. Employee complaints initiated 50.6% of events and accident reports initiated 40.1%. One third of inspections were initiated because of a fatal event, and 27.4% were initiated in response to a physical assault. Citations for security hazards were issued to 23.6% of businesses. Inspections initiated by employee complaints, in response to customer-related security issues, and involving non fatal assaults were the most frequently cited reasons for inspection. The California Division of Occupational Safety and Health conducted security inspections in a large range of industries and for diverse issues, even though no specific security codes exist. PMID- 10457508 TI - Risk indicator of dust exposure and health effects in cement plant workers. AB - A frequent practical problem of research in developing countries is the lack of reliable records on occupational hazards. To improve this situation, this article suggests and evaluates a two-phase method for estimating particle exposure. The first phase uses the focal group, or homogeneous group, technique to reconstruct the production process and estimate the level of dust exposure. The second phase applies the technique of individual history of exposure to hazards at work, an index that accumulates current and previous exposure. This method was introduced in a Portland cement plant to assess the dust-exposure levels of workers and to evaluate its usefulness in the association between estimated exposure levels and the frequency of health effects--particularly respiratory effects--that occurred as a result of such exposures. The results obtained from the analysis of the production process and of the exposure levels determined by the cement workers showed that it is possible to reconstruct the history of exposure to cement dust during each worker's occupational history. The results also showed that estimated exposure is related to respiratory damage; higher exposure resulted in more serious diseases. This supports the usefulness of the suggested methodology. PMID- 10457509 TI - Erythrocyte delta-aminolevulinic acid dehydratase genotype and other mechanisms affecting workers' susceptibility to lead. AB - In this study, the role of delta-aminolevulinic acid dehydratase (ALAD) variants in lead susceptibility was examined. The study subjects comprised 223 male workers, and the relationship between their blood lead level and erythrocyte ALAD activity or plasma/urine delta-aminolevulinic acid level was studied. Leukocyte specimens from 11 workers, whose erythrocyte ALAD activities were as low as one fifth that of the other normal workers, were subjected to analyses of their ALAD and ALAD alleles. Further, the entire exon fragment of the ALAD gene was analyzed by polymerase chain reaction, and the reaction product was used as a target for direct DNA sequencing. Genomic DNA analysis revealed that all 11 workers had the ALAD allele, whereas the entire ALAD gene analysis failed to indicate other variants, except for the Rsa I site. The depletion in erythrocyte ALAD activity was not found to be caused by the ALAD allele. PMID- 10457510 TI - Prediction of performance on the RCMP physical ability requirement evaluation. AB - The Royal Canadian Mounted Police use the Physical Ability Requirement Evaluation (PARE) for screening applicants. The purposes of this investigation were to identify those field tests of physical fitness that were associated with PARE performance and determine which most accurately classified successful and unsuccessful PARE performers. The participants were 27 female and 21 male volunteers. Testing included measures of aerobic power, anaerobic power, agility, muscular strength, muscular endurance, and body composition. Multiple regression analysis revealed a three-variable model for males (70-lb bench press, standing long jump, and agility) explaining 79% of the variability in PARE time, whereas a one-variable model (agility) explained 43% of the variability for females. Analysis of the classification accuracy of the males' data was prohibited because 91% of the males passed the PARE. Classification accuracy of the females' data, using logistic regression, produced a two-variable model (agility, 1.5-mile endurance run) with 93% overall classification accuracy. PMID- 10457511 TI - The effect of smelter work on fecundity. AB - Time to pregnancy (TTP) has been proposed as a screening instrument for the detection of environmental and occupational hazards to reproduction. Our aim was to determine whether reduced fecundity could be found among smelter workers and couples living near the smelter. The number of Months required to achieve a pregnancy was determined via a questionnaire, and TTP was analyzed using a logistic regression model with a cutoff at 12 months. No significant findings were found for a risk of having to wait for more than 12 months to achieve pregnancy because of occupational or environmental exposure. It could be that the exposure variables tested were not sensitive enough in detecting hazards to reproduction or that they were of minor importance. A tendency toward a prolonged TTP was found for couple living near the smelter. PMID- 10457512 TI - Distinctive emergency department usage for injury for workers' compensation cases in Utah in 1996. AB - To compare emergency department services paid by worker's compensation (WC) with services paid by other payers, a state database of 72,747 emergency department visits for injured adults (ages 21 to 54) in 1996 in Utah was analyzed. WC visits accounted for 21.6% (15,704) of all adult injury visits. The mean emergency department charge for WC visits was $282, and the admission rate was 17 per 1000 visits. The mean charge for other payers was $334, and the admission rate was 43 per 1000 visits. Differences were also found between these groups for Injury Severity Scores and diagnoses. In summary, WC emergency department usage was associated with less severe injuries than was emergency department usage for other payers in Utah in 1996. PMID- 10457513 TI - Use of the alkaline comet assay to monitor DNA damage in technicians exposed to low-dose radiation. AB - The exposure of human beings to ionizing radiation is still of great concern in occupational and environmental medicine, and the widespread use of radiotherapy in the treatment of cancer has led to anxiety about the possible hazards to staff who are at risk of such occupational exposure. In this study, DNA damage in the peripheral lymphocytes of 30 technicians employed in radiation oncology departments for at least 1 year were examined by the alkaline single-cell gel electrophoresis "comet" technique. The results were compared with those of 30 controls with comparable age, sex, and smoking habits who were not working in radiation oncology or chemotherapy services. The DNA damage observed in the lymphocytes of the technicians was significantly higher than that in the controls (P < 0.001). Cigarette smoking was also related to increases in DNA damage, and a significant association was found between the duration of occupational exposure to low-dose ionizing radiation and the DNA damage. PMID- 10457514 TI - Fatal harmful substances or environmental exposures in agriculture, 1992 to 1996. AB - Data from the Census of Fatal Occupational Injuries surveillance system from 1992 through 1996 were analyzed to allow a better understanding of exposures to harmful substances or environments that resulted in agricultural work fatalities. There were 357 fatalities as a result of these exposures in the agriculture production and agriculture services sectors, representing 10% of all work-related deaths that occurred in these industry sectors during this period. Contact with electric current represented 52.9% of these fatalities. Agricultural services reported 87 electrocutions, 50 of which occurred among tree trimmers. The events most likely to result in fatalities were contact with overhead power lines (26.3%) and drowning (17.1%). The overall fatality rate was 2.1 deaths per 100,000 workers. The development of appropriate hazard-awareness training for workers, such as that for electrical and drowning-related hazards, may help prevent future deaths in these industry sectors. PMID- 10457515 TI - Developing occupational and environmental medicine curricula for primary care residents: project EPOCH-Envi. Educating Physicians in OCupational Health and the Environment. AB - To help primary care residency programs develop or improve residency curricula in occupational and environmental medicine, the National Institute for Occupational Safety and Health launched a train-the-trainer initiative. This project was called EPOCH-Envi (Educating Physicians in OCcupational Health and the Environment). From 1990 to 1996, 46 2-day curriculum development workshops were held. These featured (1) guidelines on how to plan, implement, and evaluate a curriculum, (2) continuing education on occupational illnesses and injuries, (3) a worksite or environmental site visit, and (4) information resources. A total of 435 faculty from 305 residency programs participated, representing 42.5% of the family practice residencies and 24.9% of the internal medicine residencies in the United States. A survey conducted among attendees (60.4% response rate) 17 months after their workshop revealed that 65.6% of respondents had added lectures on occupational and environmental topics to the residency curriculum. Other curriculum improvements were also made. Primary care physicians manage most patients with occupational and environmental health problems or concerns. Providing technical assistance specifically designed to support occupational and environmental health education in primary care residencies can have a positive impact on curriculum content. PMID- 10457516 TI - [Evidence-based medicine--a critical overview]. AB - BACKGROUND: Evidence-based medicine (EBM) is a strategy for the standardization of medical decision-making. The purpose of this review was to demonstrate the most important tools of EBM and to critically appraise them. METHODS: The current literature concerning the subject is reviewed. A clinical example for the systematic use of EBM in diagnostic tests is explained in detail. RESULTS: The risk of an increasing bureaucratic hindrance of medical procedures by EBM is weighed against the advantages of a systematic strategy for the valid assessment of medical information. According to the clinical example EBM proves to be a valuable tool in the quantitative estimation of residual risk and reliability of diagnostic tests in everyday clinical practice. CONCLUSION: The critical use of the systematic tools of EBM for clinical decision-making is recommended. PMID- 10457517 TI - [Special characteristics of expert assessment of olfactory and taste disorders]. AB - BACKGROUND: The number of medical expert opinions dealing with smell and taste disorders has continuously increased in recent years. However, an overview of the specific problems and results of those expert opinions has not been published until now. INDIVIDUALS AND METHODS: Olfaction was assessed by multiple subjective tests in 145 individuals with chemosensory dysfunction caused by trauma or occupational and environmental exposures. Additionally, a gustatory test, nasal endoscopy, anterior rhinomanometry, and radiography of the paranasal sinuses were performed. Malignering was diagnosed on a trial of multiple symptoms. Retronasal olfaction was tested using the Guttich gustatory olfaction test. RESULTS: Most of the medical expert opinions have been ordered by professional associations (42%) and insurance companies (28%). Compensation for olfactory dysfunction was claimed after trauma (64%), occupational exposure (23%) rhinosurgical procedures (8%), and laryngectomy (5%). Anosmia or hyposmia was claimed by 66% and 24% of all individuals, respectively. According to the results of the chemosensory tests, 41% of patients suffered from anosmia and 40% had hyposmia. Malignering was registered in 14%. CONCLUSIONS: The study shows that the "characteristic case" is a male in the late fourties, complaining of anosmia initially noticed after an occipital or frontobasal head trauma. The assessment of olfactory deficits related to toxic or chemical occupational exposure is difficult when a latency between the occupational exposure and the onset of chemosensory dysfunction is present. Moreover, interactions between the occupational exposure and nicotine or alcohol abuse must be taken into consideration. Claims for medical liability arose in 3% of the study group after septoplasty and sinus surgery. PMID- 10457518 TI - [Immunohistochemical study of the distribution of constitutive nitric oxide synthase in vascular endothelium of the nasal mucosa in the human]. AB - BACKGROUND: Human nasal mucosa is a highly regulated tissue that performs a wide range of physiological functions. In addition to the classic and peptidergic neurotransmitters, the endogenously produced free radical gas nitric oxide (NO) has been found to be increasingly important for the vascular regulation of this tissue. NO-dependent control of vascular tone works in two ways, consisting of neurally produced NO acting as a neurotransmitter on the one hand and endothelially produced NO on the other hand. Neurons and endothelial cells contain various isoforms of the enzyme nitric oxide synthase, which forms NO out of L-arginine. The aim of this study was to examine the distribution of endothelial constitutive NO-synthase (ecNOS) in the human nasal mucosa of inferior turbinates. METHODS: Immunocytochemistry (avidin-biotin method) with polyclonal and monoclonal antibodies against eNOS and cluster of differentiation 31, a marker for endothelial and certain blood cells, was used in order to gain more detailed information on the physiological distribution and significance of NOS in vascular endothelium of different vessel types. RESULTS: Positive eNOS immunoreactions were found in the endothelial cells of arterial blood vessels of different diameters as well as in capillaries and postcapillary venules. Venous sinuses with or without subendothelial cushions did not show any immunoreactions. CONCLUSIONS: There is strong evidence that vascular tone in human nasal mucosa is not only subject to nerval control, but also influenced directly by mediators released from the endothelium. The present results lead to the conclusion that in physiological conditions endothelially produced NO has an influence on the arterial component of the swelling mechanism in human nasal mucosa. Because eNOS could also be detected in capillaries and postcapillary venules, NO might also play an important role in plasma extravasation. PMID- 10457519 TI - [Effect of cytokines and fibroblasts on eosinophilic granulocyte survival in polyposis nasi]. AB - BACKGROUND: Eosinophilic infiltration is the most prominent histologic feature of nasal polyps. The effects of eosinophil-associated cytokines and fibroblast derived factors on survival of isolated eosinophils from blood of patients with nasal polyps were investigated. METHODS AND PATIENTS: Peripheral blood eosinophils of 9 patients with nasal polyps were incubated in 1. culture medium alone and in a medium containing, 2. IL-5 (10 pg/ml); 3. IL-3, IL-5, GM-CSF (10 pg/ml); 4. supernatants of fibroblasts cultured from polyps of the same 9 patients; and 5. supernatants of fibroblasts enriched with IL-5, -3, and GM-CSF. Viability of eosinophils was assessed by their ability to take up WST-1. RESULTS: The survival index (T50) of eosinophils cultured in different culture medium is listed as follows: 1. medium alone: 0.8 +/- 0.14 days (MV +/- SEM), 2. medium containing IL-5; 1.2 +/- 0.19 days, 3. medium containing IL-3, IL-5, and GM-CSF: 3.1 +/- 0.13 days, 4. fibroblast supernatants: 1.6 +/- 0.29 days, and in 5. fibroblast supernatants enriched with IL-3, IL-5 and GM-CSF: 2.2 +/- 0.21 days. CONCLUSION: IL-5, prolongs eosinophil survival in vitro, IL-3, IL-5 and GM-CSF revealed synergistic effects; survival of eosinophils was significantly prolonged in comparison to stimulation with IL-5 alone. Supernatants of fibroblasts also prolonged eosinophil survival. This effect is probably due to GM-CSF, which was found in high concentrations in fibroblast supernatants. PMID- 10457520 TI - [Evaluation of nasolacrimal duct function in chronic paranasal sinus infections with 99mTc dacryoscintigraphy]. AB - BACKGROUND: Paranasal sinus infections and nasolacrimal duct obstructions are commonly encountered problems. However the relationship between these two conditions is not yet clear. Trauma, surgery, tumors, and systemic diseases are among the causes of acquired nasolacrimal duct obstructions but most of the cases are idiopathic. In this study patients diagnosed with chronic paranasal sinus infection were examined with 99mTc lacrimal dacryoscintigraphy to assess their nasolacrimal duct function. METHODS: Twenty-four patients diagnosed with chronic paranasal sinus infection are included in the study. These patients were observed for at least three months awaiting surgical treatment following unsuccessful medical treatment. The control group included 16 persons admitted to the ophthalmology department without any nasolacrimal duct disease. Nasal endoscopy and paranasal sinus CT revealed normal findings. The age of the patients with chronic paranasal sinus infection and examined in regard to nasolacrimal duct function was between 15 to 17 with a mean age of 34.2 years. Fourteen patients were male and 10 patients were female. The control group included 9 male and 7 female patients between 18 to 60 years with a mean of 30.5 years. RESULTS: The dacryoscintigraphic examination of 48 nasolacrimal ducts in 24 patients with chronic paranasal sinus infection revealed 7 complete and 18 partial obstructions of the duct, and 23 normal findings. In the control group with 32 nasolacrimal ducts of 16 patients, there was no complete obstruction. Partial obstruction (10.6, 12.0, 14.7 minutes) was be observed in three cases. The statistical workup resulted in a significant difference between paranasal sinus patients and control group (x2 = 15,840 p < 0.001). The paranasal sinus CTs of the patients with paranasal sinus disease were staged using the Lund-Mackay staging system: There is no correlation between the staging score of the paranasal sinus infection and the degree of the obstruction the nasolacrimal duct. CONCLUSION: This study shows that chronic paranasal sinus disease plays an important role in the pathogenesis of nasolacrimal duct obstructions. The presence of infection has an influence on the nasolacrimal drainage system, but the distribution (staging) of the infection does not appear to correlate with the degree of nasolacrimal duct obstruction. Partial obstruction cases that may progress to complete obstruction may be detected by dacryoscintigraphy, and the treatment of chronic paranasal sinus disease may help to overcome the lacrimal drainage problems in these cases. PMID- 10457521 TI - [Recent experimental and clinical findings retarding an interlabyrinthine connection]. AB - BACKGROUND: In his experiments on the origin and site of formation of perilymph, which he conducted using radioactive substances in 1961. Schreiner also came across a surprising secondary finding, namely that rapid substance exchange takes place between the perilymph spaces of both inner ears. This rapid transition was only demonstrated for real solutions, such as low-molecular radioactive sodium phosphate, whereas it was greatly delayed for C14-labeled amino acids. No patency was demonstrated for particles which approach the limit of visibility under a light microscope, such as cell granules consisting of radioactively labeled mitochondria from rabbits. The graded patency between perilymph spaces in relation to the molecule size of the radioactive substances added indicates a direct connection between the perilymph spaces of both inner ears. This interlabyrinthine connection first postulated by Schreiner was controversial at the time but was soon confirmed by several authors in Germany and abroad and has been known since then as the "Schreiner effect" (1964). CLINICAL FINDINGS: In spite of these publications, the hypothesis of an interlabyrinthine connection was ignored for over 20 years until the American authors Harris et al. took up the topic once again in 1985, because of clinical investigations. They termed this phenomenon "sympathetic cochleo-labyrinthitis" in line with sympathetic ophthalmia. In pathogenetic terms, these authors suspect autoimmunological genesis. It has not been clarified which anatomical and immunological routes are taken to affect the labyrinths. Possibilities being discussed include connections via the perineural lymph sheath, NEW METHODS: In recent times, the perivascular spaces have also been considered, and Maher et al. (1997) attempted to substantiate this theory by demonstration using contrast media. In 1997, W. Zenker, the Zurich anatomist, conducted experiments using ferritin and was the first to demonstrate movement of such molecules within the dura. He believes that the transition of substances from one inner ear to the other is via CSF movements in the subarachnoid space and in the perivascular compartment of numerous small vessels crossing in a median plane such as veins of the clivus and plexus cavernosi but also venules and arterioles. PMID- 10457523 TI - [The "ear fever thermometer"--studies of ear thermography]. AB - BACKGROUND: Though rarely mentioned in the ENT literature, the "ear thermometer" has become more and more popular in recent years, not only in hospitals but also in households. These instruments are easy to use, and their infrared technology is said to provide precise measurements. The purpose of this study was to verify these claims. PATIENTS AND METHODS: Infrared thermometers were tested in various conditions, and the results were compared. We assessed the effects of "distending" the outer ear canal, different body positions, and irritation of the ear (slight external otitis, hearing aids, otitis media, etc.). We did not specifically test effects of ear wax, as it had sufficiently been studied in pediatric or anesthesiological papers before (almost no effect, except in cases of occlusion of the auditory canal). RESULTS: Using the ear thermometer we found small but statistically significant differences in febrile patients in different body positions. Irritated ears always showed higher temperatures than the normal contralateral ears. The most significant differences were found in persons lying on one side. The "pillow ear" was found 0.7 degree C (average) warmer than the contralateral ear. CONCLUSIONS: Ear thermometers for estimating the body temperature permit easy and fast measurements. However, they include possible sources of measurement error. This study describes possible errors that the therapists should be aware of to avoid misinterpreting the course of a disease. PMID- 10457522 TI - [Surgical treatment of acoustically-induced vertigo (Tullio phenomenon)]. AB - BACKGROUND: Tullio phenomenon is defined as noise induced vertigo. Other authors have attributed this symptom to either a perilymphatic fistula or postinflammatory adhesions between the stapes foot plate and the vestibular end organs. METHOD: In this paper two cases are described in which acoustically induced vertigo was explained by abnormal mobility of the stapes. RESULTS: The stapes was stabilized by the placement of cartilage chips beside the crurae of the stapes. Both cases demonstrated long term success, i.e. 4 and 5 years postoperatively. PMID- 10457524 TI - [Congenital laryngotracheo-esophageal cleft: diagnosis and surgical treatment by anterior, translaryngeal approach]. AB - BACKGROUND: Laryngotracheoesophageal cleft is a rare but potentially life threatening anomaly. Less than 200 cases have been published to date. Both the diagnostic and therapeutic recommendations are discussed controversially in the international literature. PATIENT: We report on the diagnostic and surgical management of a type III cleft larynx in a one month old male presenting with aspiration, pneumonia, and aphonia. Hypoplasia of the cricoarytenoid muscles was associated to the cleft. Rigid endoscopy was found to be the best tool for the diagnostic exploration of the cleft, whereas flexible endoscopy failed to detect the defect. The cleft was broadly exposed using a modified anterior translaryngeal approach that included a tracheostomy. After debriding the mucosal margins, the defect was closed in two layers, and a t-shaped Montgomery tube was implanted. Two further revisions using the mentioned translaryngeal approach and one endoscopic procedure were necessary to achieve complete and permanent closure of the cleft. RESULTS: Twenty months after birth the boy is able to swallow thick and liquid food without any problems. Stable scar tissue has grown within the former cleft region. The vocal cords are somewhat thickened but mobile in a reduced range. Both the trachea and the esophagus show quite normal diameters. CONCLUSION: Considering the fact that the arytenoid cartilages touch or overlap each other a congenital defect within the posterior midline of the larynx can only be diagnosed by rigid endoscopy that spreads the cleft apart. In addition to our positive experiences with traumatic fistulas and stenosis of the juvenile trachea we recommend now the anterior vertical laryngeal incision for the operative management of the congenital type III cleft larynx. This direct open approach provides excellent exposure of all components of the defect without the risk of recurrent laryngeal nerve injury. Subglottic stenosis or impaired stability of the larynx described by other authors did not occur in this case. However, the postoperative period is relatively short and careful follow-up for a period of several years is therefore required. PMID- 10457525 TI - [Pseudoaneurysm of the lingual artery after tonsillectomy. A rare complication]. AB - BACKGROUND: Tonsillectomy is one of the most common surgical procedures in otorhinolaryngology; however, life-threatening complications can occur in rare cases. PATIENTS: We report about a seven-year-old girl who developed a pseudoaneurysm of the lingual artery following elective routine tonsillectomy. RESULTS AND CONCLUSION: Initial ENT examination will reveal a painful cervical mass or unilateral palatal swelling that can easily be misinterpreted as a peritonsillar (or parapharyngeal) abscess. Computed tomography and angiography are indicated to confirm the diagnosis. Bleeding from a pseudoaneurysm of any branch of the external carotid artery represents a serious and life-threatening complication. Surgery is the treatment of choice. PMID- 10457526 TI - [Postoperative hemorrhage after tonsillectomy]. PMID- 10457527 TI - [Recommendations for medical documentation in suspected occupationally-induced hearing loss]. PMID- 10457528 TI - [Interesting case no. 26. Disruption of the ossicular chain--an interesting differential otosclerosis diagnosis]. PMID- 10457529 TI - [Surgical voice rehabilitation after laryngectomy. II: Secondary voice rehabilitation]. PMID- 10457530 TI - Olfactory dysfunction in schizophrenia: a qualitative and quantitative review. AB - Olfactory dysfunction in patients with schizophrenia has been a topic of increasing interest, with deficits in odor identification, detection threshold sensitivity, discrimination, and memory being reported. Despite increasing knowledge, controversy has existed about possible differential deficits among olfactory tests as well as the influences of gender, smoking, and medication status on olfactory measures. To help elucidate some of this controversy, we conducted a qualitative and quantitative (meta-analytic) review of the English language literature on olfaction in schizophrenia. Moderator variables such as gender, medication status, and smoking history were also examined. Results indicated that substantial olfactory deficits, across all domains, are observed in patients with schizophrenia. No differential deficits were observed across domains of odor identification, detection threshold sensitivity, discrimination, and memory. The influences of gender, medication status, and smoking on effect sizes were not significant across studies. This supports the hypothesis of primary dysfunction in the olfactory system that is regulated by brain regions where structural and functional abnormalities have also been reported in neuroimaging studies. PMID- 10457531 TI - Ionotropic glutamate receptor modulation of 5-HT6 and 5-HT7 mRNA expression in rat brain. AB - The novel serotonin receptor subtypes, 5-HT6 and 5-HT7, are located in limbic regions and have nanomolar affinities for atypical antipsychotics. These factors have led some to speculate about the involvement of 5-HT6 and 5-HT7 receptors in schizophrenia. However, relatively little is known about these receptor subtypes, including the regulation of their expression in limbic regions. In particular, the regulation of extracellular serotonin levels in the striatum and hippocampal formation by glutamate receptors led us to examine the effects of systemic ionotropic glutamate receptor modulator treatment on 5-HT6 and 5-HT7 receptor expression in these regions. MK-801 treatment induced a dose-dependent decrease in striatal 5-HT6 receptor mRNA levels; similarly, both aniracetam and NBQX treatments also led to decreases in striatal 5-HT6 receptor mRNA levels. Hippocampal 5-HT6 and 5-HT7 receptor expression were not dramatically affected by any of the treatments. To our knowledge, this is the first demonstration of the regulation of striatal 5-HT6 receptor mRNA expression, and provides neurochemical anatomical evidence for the interaction of serotonergic and glutamatergic systems. Furthermore, although these two neurotransmitter systems are separately implicated in schizophrenia, the glutamatergic regulation of the expression of a receptor subtype associated with schizophrenia suggests that alterations in serotonin receptor expression in schizophrenia may result, in part, from altered glutamatergic activity. PMID- 10457532 TI - Effects of antidepressants on 5-HT7 receptor regulation in the rat hypothalamus. AB - Recent evidence suggests that a novel serotonin receptor 5-HT7 localized in the hypothalamus downregulates in response to treatment with the antidepressant fluoxetine (Sleight et al. 1995). This receptor has also been implicated in the regulation of circadian rhythms (Lovenberg et al. 1993). Here, we show that several agents administered in a profile consistent with activity at the 5-HT7 receptor produce significant functional Fos immunoreactivity in the suprachiasmatic nucleus (SCN), an effect reduced upon chronic exposure. Furthermore, binding studies demonstrate that chronic administration of Fos inducing agents produces a neuroadaptive downregulation of the 5-HT7 receptor in the hypothalamus. The current studies extend the previous observations to include several pharmacologically distinct antidepressants. In addition, these studies provide further evidence to support the role of the 5-HT7 receptor in the mechanism of antidepressant action and in the regulation of circadian rhythms controlled by the SCN. PMID- 10457533 TI - Global cerebral blood flow increase reveals focal hypoperfusion in schizophrenia. AB - Recent functional neuroimaging strategies have evaluated cerebral blood flow (CBF) to determine specific sites of action of pharmacologic agents. Since many pharmacologic agents change global CBF, we investigated the effects of global CBF changes on regional perfusion with acetazolamide, which increases global CBF via non-neuronal mechanisms. We used the [15O]PET technique to measure CBF before and after we infused 8 schizophrenic patients and 10 healthy control subjects with acetazolamide. The rostral anterior cingulate cortex demonstrated a greater perfusion increase in the schizophrenic subjects after acetazolamide infusion, relative to other areas of the brain. During the baseline condition, this area showed relative hypoperfusion in our sample of schizophrenic subjects, consistent with previous functional neuroimaging studies. The results demonstrate the need for caution in interpreting CBF changes after pharmacologic challenge, because global CBF changes can confound the assessment of regionally-specific pharmacologic action. PMID- 10457534 TI - Mitochondrial function is differentially altered in the basal ganglia of chronic schizophrenics. AB - In the present study, we have applied a novel strategy involving the postmortem measurement of the mitochondrial respiratory chain enzyme cytochrome-c oxidase (COX; complex IV) to identify regional changes in energy metabolism in the basal ganglia of chronic, medicated schizophrenics. COX activity was decreased in the caudate nucleus but increased in the putamen and nucleus accumbens. An increase in succinate dehydrogenase (complex II) was evident in the putamen and nucleus accumbens, but changes were not seen with NADH dehydrogenase (complex I). An analysis of interregional correlations in energy metabolism revealed several anomalies in the connections between the caudate and putamen and the globus pallidus in schizophrenics. Results provide strong evidence that changes in baseline energy metabolism in specific regions of the basal ganglia may exist in the disease. Based upon the high degree of input it receives from associative cortical areas, results suggest that a defect in the caudate may underlie certain aspects of cognitive decline in schizophrenics. In contrast, an increase in COX in the putamen, which receives extensive projections from the sensorimotor cortex, may reflect an effect of chronic neuroleptic treatment on motor function. PMID- 10457535 TI - Changes in auditory selective attention and event-related potentials following oral administration of D-amphetamine in humans. AB - The effect of d-amphetamine on selective attention in humans was investigated by measuring event-related potentials (ERPs) during a complex auditory selective attention task (CSAT). The CSAT required subjects to make a button press response to infrequent target tones presented amongst tones that varied in pitch (high vs. low), location (left vs. right ear) and duration (51 ms vs. 102 ms). Healthy subjects completed the CSAT under three conditions: placebo, 10 mg and 20 mg d amphetamine, at least one week apart. D-amphetamine produced a significant dose response increase in hit-rate and decrease in reaction time without changing false alarm rate. D-amphetamine reduced late PN to location irrelevant stimuli and pitch irrelevant stimuli in both the attended and unattended location. The effect of d-amphetamine was interpreted as a decrease in the maintenance of the attentional trace to irrelevant stimuli. However, these changes were accompanied by some evidence of processing of stimulus features in the unattended location. These results suggest that d-amphetamine improves selective attention, and decreases the maintenance of attention to irrelevant stimuli. PMID- 10457536 TI - Electrical stimulation of rat medial prefrontal cortex enhances forebrain serotonin output: implications for electroconvulsive therapy and transcranial magnetic stimulation in depression. AB - Decreased activity of the prefrontal cortex (PFC), as well as reduced serotonergic neurotransmission, is considered as a characteristic feature of major depression. The mechanism by which electroconvulsive therapy (ECT) and transcranial magnetic stimulation (TMS) achieve their antidepressant effects may involve changes in PFC activity. It is, however, still unclear whether these changes are accompanied by increased synaptic availability of serotonin (5-HT). In the present study, 5-HT efflux in the rat ventral hippocampus and amygdala was analyzed using in vivo microdialysis during low-current electrical stimulation of PFC and other cortical regions. Electrical stimulation of the medial PFC produced current-dependent increases in limbic 5-HT output in both urethane-anesthetized and behaving rats. No effects on 5-HT levels were seen after comparable stimulation of either the lateral parts of the PFC, the medial precentral area, the primary motor cortex or the parietal cortex. This pronounced regional specificity of the effect of medial PFC stimulation on limbic 5-HT output suggests that activation of this particular area might play a crucial role in such antidepressant treatments as ECT and TMS. PMID- 10457538 TI - Melatonin sensitivity to dim white light in affective disorders. AB - Both dim and bright light has been shown to suppress the nocturnal secretion of the pineal hormone melatonin. Early reports suggests that an abnormal response to light occurs in patients with bipolar affective disorder, where as patients with major depressive disorder respond similarly to controls. It has been suggested that this abnormal sensitivity of the melatonin response to light could be a trait marker of bipolar affective disorder. However reports lack consistency. Hence, we investigated the melatonin suppression by dim light (200 lux) in patients with bipolar affective disorder, seasonal affective disorder and major depressive disorder. Results suggest that a supersensitive melatonin suppression to light in bipolar affective disorder (p < .005), and seasonal affective disorder (p < .05), whereas patients with major depressive disorder display similar suppression to controls. The supersensitivity may be a mechanism where by phase-delayed rhythms, are resynchronised to a new circadian position. Conversely, an abnormality may exist in the pathway from the retina to the suprachiamatic nucleus. PMID- 10457537 TI - Glucocorticoid effects on mesotelencephalic dopamine neurotransmission. AB - Multiple neurochemical estimates were used to examine peripheral corticosterone (CORT) effects in dopaminergic terminal regions. Acute CORT administration, which elevated plasma CORT (5 h), slightly decreased dihydroxyphenylacetic acid (DOPAC) to dopamine (DA) ratios in the striatum but not in other regions examined. Two weeks of adrenalectomy (ADX) increased both medial prefrontal cortex DOPAC/DA and homovanillic acid (HVA)/DA and striatal HVA/DA. A reciprocal pattern of changes was observed with CORT replacement in ADX animals. In contrast, CORT replacement in ADX animals did not significantly influence tyrosine hydroxylase content, basal dihydroxyphenylalanine (DOPA) accumulation after NSD 1015 treatment or the decline in DA after alpha-methyl-para-tyrosine, suggesting that neither DA neuronal activity nor release are altered by CORT. Moreover, neither gamma hydroxybutyric acid lactone-induced increases in DOPA accumulation or stress induced increases in DA utilization were influenced by CORT replacement, indicating that neither autoreceptor regulation of DA synthesis nor acute stress regulation of DA utilization are changed by CORT. The findings are most consistent with direct inhibition of basal DA metabolism in the medial prefrontal cortex and striatum. The possible physiological and behavioral significance of this inhibition is being further explored. PMID- 10457539 TI - A behavioral analysis of the spatial learning deficit induced by the NMDA receptor antagonist MK-801 (dizocilpine) in the rat. AB - This study analyzes whether the disruptive effects of the noncompetitive NMDA receptor antagonist MK-801 (0.01-0.1 mg/kg s.c.) on spatial learning can be dissociated from sensorimotor disturbances in the rat. Two different modifications of the Morris swim maze task with a hidden underwater platform were used: with or without local cue. Retention was tested either 24 h or 7 days after training as a probe trial (without platform). The present data indicate that MK 801 produces an impairment of spatial learning that cannot be dissociated from motor or sensory mechanisms. These findings support the view that NMDA receptors probably contribute to, but are not essential for, spatial learning in the water maze. PMID- 10457540 TI - Effects of repeated high-dose methamphetamine on local cerebral glucose utilization in rats. AB - Repeated administration of high doses of methamphetamine (MAP) to rats can induce long-lasting neurotoxicity which may be related to permanent psychotic symptoms and negative symptoms in some MAP psychotic patients. In this study, we used the 2-[14C]deoxyglucose (2DG) method to analyze the effects of repeated MAP administration (12.5 mg/kg, i.p., 4 times every 2 hr within a day) 14 days and 60 days after drug administration. The results showed a widespread (26 of the 43 regions examined) decreases in the regional cerebral glucose utilization. The regions with decrease metabolism included all the extrapyramidal systems, the hippocampus formation and dorsal raphe nucleus. Rats tested 60 days after drug administration has similar finding to those with a 14-day abstinent period. The results of the functional change in this study provide support for the neurotoxic effects of repeated high dose MAP administration in rats. Furthermore, the neurotoxic effects are selective and long-lasting. We suggested the MAP neurotoxic model can be used to study the permanent psychosis and negative symptoms of MAP-induced psychosis in humans. PMID- 10457541 TI - Effects of naltrexone and fluoxetine on alcohol self-administration and reinstatement of alcohol seeking induced by priming injections of alcohol and exposure to stress. AB - We have recently shown that priming injections of alcohol and footshock stress reinstate alcohol seeking in drug-free rats. Here we tested whether naltrexone and fluoxetine, two drugs used in the treatment of alcohol dependence, would affect reinstatement of alcohol seeking induced by these events. We also determined the effects of these drugs on alcohol self-administration during the maintenance phase. Rats were trained to press a lever for a 12% w/v alcohol solution. After stable drug-taking behavior was obtained, lever pressing for alcohol was extinguished. Reinstatement of drug seeking was then determined after priming injections of alcohol (0.24-0.96 g/kg) or exposure to intermittent footshock (5 and 15 min). Rats were pretreated with naltrexone (0.2-0.4 mg/kg) or fluoxetine (2.5-5 mg/kg) during maintenance or during tests for reinstatement. Both naltrexone and fluoxetine decreased lever presses for alcohol during the maintenance phase. Naltrexone blocked alcohol-induced, but not stress-induced reinstatement. In contrast, fluoxetine blocked stress-induced reinstatement, while its effect on alcohol-induced reinstatement was less consistent. The implications of these data to the understanding of relapse to alcohol are discussed. PMID- 10457542 TI - Limited sex differences in response to "binge" smoked cocaine use in humans. AB - The subjective and physiological effects of repeated smoked cocaine self administration were compared in 11 men and 9 women. Twice a day, on 2 consecutive days, participants smoked up to six 50-mg doses of cocaine base, at 14 min intervals. Men and women self-administered a similar number of cocaine doses (21.7 and 21.6, respectively). The most striking sex difference was that women had higher cocaine plasma concentrations than men (632.7 ng/ml vs. 376.7 mg/ml) after the sixth cocaine dose of the first session. After the first cocaine dose, women reported that they would spend significantly less for the dose than men ($1.58 vs. $3.15). Although cocaine produced similar effects in men and women 4 min after each dose, 15 min after the last dose of the session, heart rate and blood pressure remained elevated in women, but ratings of "I want cocaine" were lower in women as compared to men. Thus, smoking cocaine produced similar acute subjective effects in men and women, but prolonged cardiovascular effects and higher cocaine plasma concentrations in women. PMID- 10457543 TI - Serotonin-2 receptors and human sleep: effect of a selective antagonist on EEG power spectra. AB - To investigate the effect on the sleep EEG, a 1-mg oral dose of SR 46349B, a novel 5-HT2 antagonist, was administered three hours before bedtime. The drug enhanced slow wave sleep (SWS) and reduced stage 2 without affecting subjective sleep quality. In nonREM sleep (NREMS) EEG slow-wave activity (SWA; power within 0.75-4.5 Hz) was increased and spindle frequency activity (SFA; power within 12.25-15 Hz) was decreased. The relative NREMS power spectrum showed a bimodal pattern with the main peak at 1.5 Hz and a secondary peak at 6 Hz. A regional analysis based on bipolar derivations along the antero-posterior axis revealed significant 'treatment' x 'derivation' interactions within the 9-16 Hz range. In enhancing SWA and attenuating SFA, the 5-HT2 receptor antagonist mimicked the effect of sleep deprivation, whereas the pattern of the NREMS spectrum differed. PMID- 10457544 TI - Psychopathy as a risk factor for violence. AB - As a result of Kansas v Hendricks, many sex offenders in the U.S. are likely to be civilly committed to mental institutions for indefinite periods, and many others with histories of violent offenses may also be so committed. It therefore becomes critical for mental health professionals to understand the risk factors for re-offending that put the public in jeopardy. The most reliable of these factors is psychopathy, which will here be defined, along with its differentiation from the more commonly diagnosed antisocial personality disorder. The assessment of psychopathy, its relationship to crime--especially, to violent crime, its (non-) responsiveness to the usual treatment, and an outline of a potentially more effective one, are presented. Finally, and particularly in view of its widely accepted validity, the potential for abuse of the PCL-R and :SV are noted. PMID- 10457545 TI - The aggressive patient/inmate: beyond denial. AB - The public is no longer willing to tolerate being repeatedly victimized by a small group of sexually and/or physically aggressive predators. These predators may be found among much larger groups of non-predatory inpatients in psychiatric hospitals or inmates in prisons. These institutionalized people deserve to be treated or incarcerated without fear of being victimized, as well. The U.S. Supreme Court has recently recognized the principle of protecting the public from such predation, in its Kansas v Hendricks decision. This paper details the philosophy, management, unit construction, legislative changes, and intervention strategies necessary to contain such predators and to propel them toward rehabilitation-once we have given up our denial about the existence of such people. PMID- 10457546 TI - The paraphilias, obsessive compulsive spectrum disorder, and the treatment of sexually deviant behaviour. AB - The paraphilias have been mostly ignored by psychiatry, even though psychiatrists are ideally suited to treat and diagnose these disorders by virtue of their medical and psychological training. The sexual deviations require an understanding of both biological and psychological causation and skills in psychological and pharmacological treatments. More recently the Supreme Court of the United States in Kansas v Hendricks (1997) upheld the constitutionality of the civil commitment of sexually deviant individuals for psychiatric treatment. As the various states adopt statutes based on Hendricks, psychiatry will be forced to take an active interest in the diagnosis and the management of the paraphilias. This paper outlines briefly where the field is in the understanding of the natural history, diagnosis, and treatment of the paraphilias. PMID- 10457547 TI - The "mad" vs. the "bad" revisited: managing predatory behavior. AB - Ever since the Englightenment, western democracies have maintained separate institutions for the "mad" and for the "bad." For the mad they provided protection and treatment, and for the bad, deterrence and punishment. During the past decade, however, increasing social conservatism and the apparent failure of the criminal justice system have resulted in a series of changes in social policy. In order to protect the public adequately from repeatedly violent predators, legislation has been introduced in several states that remands those predators to mental institutions on completion of their prison terms. Challenges to this legislation, based on ex post facto, double jeopardy, and civil rights considerations were effectively dismissed by the U.S. Supreme Court in Kansas v Hendricks in June, 1997. An effect, however unintended, of upholding these laws and of policy changes leading up to them, is that they have effectively blurred the separation between mad and bad. This paper details the following: 1) the events that have led to these changes in policy and law; 2) the preparations being made in New York State's mental health system to prepare for managing such predators when and if such laws are enacted; 3) the dangers and drawbacks these changes have brought in states where they have already been legislatively mandated; and 4) suggestions for alternatives that would reaffirm the separation between mad and bad. PMID- 10457548 TI - Outreach workers' experiences in a homeless outreach project: issues of boundaries, ethics and staff safety. AB - Mental health professionals and researchers have emphasized the importance of conducting outreach to locate homeless persons with mental illness, and of creatively engaging these persons into a therapeutic relationship. These outreach and engagement activities raise challenging issues in the areas of client-staff boundaries, professional ethics, and staff safety. While several issues in each of these three key areas have received attention in the growing literature on homelessness, certain issues within each area remain unexplored. The authors draw from the street experiences of outreach staff in a federally funded homeless outreach project to further explore each of these areas, and suggest that experiences of outreach workers are essential in shaping and redefining work activities in these, and other important areas. PMID- 10457549 TI - Characteristics of assaultive psychiatric inpatients in an era of managed care. AB - Prior to managed care, extensive research documented the characteristics of assaultive inpatients in traditional state mental hospital settings as primarily older, male, psychotic patients with histories of violence toward others and of substance abuse. Recent early studies in rural and urban hospital settings have suggested that the characteristics of assaultive patients may be changing to include younger, more frequently female, patients with personality disorders and histories of personal victimization. This two-points-in-time study sought to assess the nature of assaultive patients in a suburban traditional state mental hospital after the implementation of managed care initiatives, and compared to the nature of the assaultive patients before and after the downsizing of this state mental health facility. Before census reduction, the assaultive patients were of the traditional type. After census reduction, the assaultive patients reflected more recent trends. The implications of the findings are discussed, and strategies for fostering facility safety in light of the newer violent patient are outlined. PMID- 10457550 TI - [C-reactive protein and transthyretin in early diagnosis of infection after open fractures of the lower limbs (a preliminary study)]. AB - PURPOSE OF THE STUDY: The authors investigated the value of C-reactive protein (CRP) and transthyretin (TTR) in the early diagnosis of infection after open fractures of the lower limb in an open, prospective study. MATERIAL AND METHODS: Eighty patients were treated with acute debridement and bone fixation. Follow-up included clinical, radiological, bacteriological and biological assessment: white cell blood count (WBC), erythrocyte sedimentation rate (ESR), CRP and TTR. Diagnosis of infection was based on macroscopic presence of pus. RESULTS: Post operative reference biological profiles were defined in 74 cases without infection. Reference profiles of WBC and ESR showed unreliable interindividual variations and could not be considered for the diagnosis of infection. Reference profiles of CRP and TTR showed a respective increase (for CRP) and decrease (for TTR) in the early post-operative course, with return to normal values after 12 days. In 6 infected patients, CRP concentrations were suddenly increased and TTR concentrations decreased at the time (3 cases) or even before (3 cases) clinical diagnosis of infection. These variations were mostly simultaneous. No unusual profile was found. The ratio of CRP/TTR concentrations experienced also a sudden increase in infected cases. DISCUSSION: Because of not specifical and unreliable variations in the post-operative outcome of non infected patients, WBC and ESR cannot be considered for the early diagnosis of infection. CRP and TTR concentrations with a respective cut-off value of 100 mg/L and 120 mg/L were found efficient for the early diagnosis of infection, and preceded clinical diagnosis in three of them. A CRP/TTR ratio over 60 p. 100, 8 days or more after initial surgery was found to be very specific (93 p. 100) and sensitive (100 p. 100) for the diagnosis of infection. CONCLUSION: Serial quantifications of CRP and TTR should be performed every four days during the follow-up of open fractures in order to early diagnose a post-operative infection. Comparison of both CRP and TTR could allow a higher accuracy, because of the possible lack of variation of one the two markers. PMID- 10457551 TI - [Infection prophylaxis in open leg fractures. Comparison of a dose of pefloxacin and 5 days of cefazolin-oxacillin. A randomized study of 616 cases]. AB - THE PURPOSE OF THE STUDY: Was to compare the efficacy of a single 800 mg injection of Pefloxacin (PF) versus 2 days of cefazolin (1 gr.Q.6 H) followed by 3 days of oxacillin (1 gr.Q.8 H) in patients with an open tibial fracture and to examine the predictive factors for infection. A double-blind double dummy, multicentric, randomized trial was performed. 616 adults with an open tibial fracture requiring single-stage bone coverage were included. The end point was wound infection within 3 months. RESULTS: Within 3 months, 21/316 patients were infected in the PF group (6.6 p. 100) versus 24/300 in the CZ-OX group (8 p. 100), the difference was not significant (95 p. 100 Cl for difference: -4.8 p. 100 to 2.1 p. 100). Twenty one strains were isolated in 18 infected patients in the PF group, and 27 in 20 patients in the CZ-OX group. Negative gram bacteria were less frequent in the PF group (10 p. 100) than in the CZ-OX group (48 p. 100), and positive gram bacteria were more frequent in the PF group (90 p. 100) than in the CZ-OX group (52 p. 100). Independent risk factors for infection were severe contamination, widespread contusion, unstable fracture, positive sample in the emergency room and at the end of surgery. Resistant infecting bacteria rate was 24 p. 100 in infected cases. CONCLUSION: There was no difference in infection rates after surgery for open tibial fractures between a 800 mg injection of Pefloxacin and 2 days of pephazolin followed by 3 days of oxacillin. Infecting bacteria were mainly nosocomially acquired. PMID- 10457552 TI - [Revision of infected total hip prostheses by ablation reimplantation of an uncemented prosthesis. 57 case reports]. AB - PURPOSE OF THE STUDY: 57 cases of infected total hip prosthesis treated by removal of the implant and implantation of unncemented prosthesis, were studied to evaluate functional and sepsis results. MATERIAL AND METHODS: 57 patients treated by reimplantation of an uncemented total hip prosthesis after removal of the infected prosthesis were observed. 16 patients underwent a single-stage exchange, 41 a two-stage reimplantation. 46 cases were analysed for infection findings (clinical, radiological and biological assessment) and only 34 cases for functional evaluation (PMA scale, Harris score) with a mean follow-up of 6.6 years. The antibiotic therapy was adapted to each patient but generally, the treatment was prolonged. RESULTS: At follow-up time (which might be too short in time), only 2 patients had a recurrence of infection. One had a single-stage exchange (reoperated by two stage exchange with a good final result at 6 years follow-up), the other a two-stage exchange. In both cases we found that postoperative antibiotic therapy was inadequate. Functional results were better with PMA scale (23 good results of 34) than with Harris score (14 excellent or good results only). 5 patients were reoperated for mechanical implant failure. DISCUSSION: Since 1991, we adopted a standardized procedure to treat chronic infected total hip prosthesis including: routine preoperative aspiration of symptomatic prosthesis; removal of the implant and around debridement followed at a later date (6 weeks) by reimplantation using uncemented implants (hydroxyapatite coated implant). Postoperative antibiotic therapy has to be massive (parenteral bitherapy for at least 21 days after each operative stage) and has to last 6 months after reimplantation. This procedure seems reliable and corroborate the validity of two-stage treatment. The using of uncemented implants allows a good bone reconstruction and does not seem to increase the risk of septic recurrence. CONCLUSION: It is quite difficult to find a hard and fast rule in infected prosthesis treatment, because many factors can influence results. The proposed procedure seems reliable, even if antibiotherapy is long and hard, but requires a strong collaboration between bacteriologist infectiologist and surgeon. PMID- 10457553 TI - [Treatment of fresh humeral diaphysis fractures by Seidel intramedullary locked nailing. A study of 23 initial cases after 2.5 years with rotator cuff evaluation]. AB - PURPOSE OF THE STUDY: The Seidel intramedullary locking nail is one of the alternative methods for operative treatment of humeral shaft fractures. Short follow-up demonstrated persistent pain and stiffness at the shoulder joint. The aims of this retrospective study were first to evaluate results with attention to operative technique, consolidation rate and postoperative complications. Second we studied rotator cuff healing in our first patients. MATERIAL AND METHODS: Twenty-five acute diaphyseal fractures were treated between May 91 and Dec 94. We reviewed personally 23 patients with a mean follow-up of 33 months (range 22-66). The dominant fracture type according to AO was type A. Clinical shoulder assessment was carried out using the Constant score. Isometric strength was recorded in both shoulders with a hand-held dynamometer in abduction, external and internal rotation. Sonographic evaluation of the rotator cuff was performed using a 7.5 MHz linear array transducer in all 23 patients. RESULTS: All but one fracture healed at an average of two months. Impingement was observed in three patients but pain relief and normal shoulder motion have been reached after nail removal. Infection occurred in one patient but final result was good. Constant score averaged 78.7 (range 51-94.2) classifying the result in all but two patients as excellent or good. Compared with the external and internal rotations, strength was significantly reduced in abduction but reached 83.5 per cent of the opposite shoulder. No statistical differences were found in relation with age, gender and side. Compared with the contralateral shoulder, rotator cuff evaluation with sonography was considered as normal in 18 patients. Calcium deposits of the cuff were noticed in the infected patient. In three cases sonography detected hyperechoic line considered as scar in the supraspinatus tendon without any partial or full-thickness tear. DISCUSSION AND CONCLUSION: A median starting point avoids the avascular area and gives a straight access to the medullary canal. This study demonstrates that using this entry portal and a reliable technique antegrade nailing of the humerus does not compromise rotator cuff healing and shoulder function. Technical errors lead to poor or fair results but despite this learning curve, Seidel nail when operative treatment is indicated, is a good choice. Attention must be paid to patients with clinical history of impingement or rotator cuff tendinopathy. PMID- 10457554 TI - [Glomus tumor of the nail area. Apropos of a series of 55 patients]. AB - PURPOSE OF THE STUDY: Glomus tumors are infrequent in the hand. We have review 55 cases, localized in the nail area to discuss their diagnosis and surgical approach. MATERIAL AND METHODS: Out of 80 glomus tumors of the hand seen in two hand units, 55 were located around the nail area, 33 in the nail bed area, 8 at the nail root and 14 laterally. The population was predominantly female (42 cases) and the mean age 43 years. The mean delay before presentation was 37 months. Pain was the main symptom (97 per cent) increased by trauma and cold. A blue spot was visible in 15 cases and diagnosis was performed on clinical ground in all cases except two. A notch was present on X-ray in 16 cases. A prospective study with MRI allow a diagnosis in 18 of 21 cases. All tumors were removed through a lateral approach elevating the nail complex and confirmed histologically. RESULTS: The diagnosis was per-operatively confirmed in 53 cases and performed in one case. The last case of supposed hemangioma was modified at histological examination. No case of nail dystrophy (not existing pre operatively) or residual pain was seen at the 81 months of follow up. Seven recurrences were observed, 4 of them having been operated initially in our units. DISCUSSION: Clinical diagnosis could be performed on clinical grounds. MRI is to be reserved to recurrences or multi-operated patients. Risk of recurrence has to be mentioned pre-operatively to the patient. CONCLUSION: Lateral approach with nail complex elevation is safe, allowing excision of the tumor without nail dystrophy. PMID- 10457555 TI - [Reconstruction of the anterior cruciate ligament: arthrotomy versus arthroscopy]. AB - The aim of this study was to compare the results of arthroscopic with open arthrotomy reconstruction of the anterior cruciate ligament, as treatment of chronic anterior laxity. 54 knees (among 63) were evaluated, 33 reconstructions were performed according to Kenneth Jones technique with arthrotomy (from 1990 to may 1993) and 21 were arthroscopically-assisted (from may 1993 to 1996). Meniscectomy was associated respectively in 22 and 7 cases. The average interval between initial injury and surgery was 18 and 18.6 months. Follow-up was one year at least. We evaluated mobility, amyotrophy and quadriceps and hamstrings muscular deficit: static at 1 month and using Cybex isokinetic tests at 2, 3 and 6 months and 1 year. Postoperative residual laxity and Arpege cotation were evaluated at 1 year. Student and Mann Whitney tests were used for statistical evaluation. As complication we noted respectively after arthrotomy and after arthroscopy: 7 (21.2 p. 100) and 4 (19 p. 100) algodystrophy, 1 (3 p. 100) and 1 (4., 7 p. 100) Cyclops syndrome, and 2 (6 p. 100) and 1 (4.7 p. 100) anterior knee pain. Lack of extension and flexion were respectively -5.4 degrees/130 degrees and -1.9 degrees/136 degrees at 3 months (p = 0.04) and -3.5/134 degrees and -1.5 degrees/138 degrees at 6 months (not significative). At 1 month, static hamstrings deficit was 41.3 p. 100 after open arthrotomy and 29.6 p. 100 after arthroscopic assisted (p = 0.05). At 2 months, isokinetic hamstrings deficit was lower after arthroscopic assisted (21.6 p. 100 at 60 degrees; 20.8 p. 100 at 180 degrees) than after open arthrotomy (32.8 p. 100; 32.5 p. 100) (p = 0.039 and 0.008). This difference was found for hamstrings until 3 months. At 6 months and 1 year, no difference was found for Cybex tests. In Arpege score, at 1 year, 73.3 p. 100 were very satisfied or satisfied after open arthrotomy and 77.7 p. 100 after arthroscopy. Global results were excellent or good in 66 p. 100 after open arthrotomy and 83 p. 100 after arthroscopy. Radiological laxity was less than 5 mm in 88 p. 100 after open arthrotomy and 92 p. 100 after arthroscopic assisted. So, after arthroscopically assisted procedure, the number of algodystrophy and anterior knee pain was lower, and until 3 months, range of motion was better and hamstrings deficit was lower. After 6 months, difference about range of motion or muscular deficit were not significative. At 1 year, after arthroscopic procedure, results seemed better with a lower rate of residual laxity and better global results, but the number of medial meniscectomies was lower in this group. In conclusion, the arthroscopic-assisted procedure seems to allow a faster rehabilitation. PMID- 10457556 TI - [Hallux valgus treated by Scarf osteotomy of the first metatarsus and the first phalanx associated with an adductor plasty. Apropos of 50 cases with a 2-year follow up]. AB - PURPOSE OF THE STUDY: Fifty hallux-valgus were treated with Scarf Osteotomy of the first metatarsal, associated to a phalangeal varisation or shortening osteotomy and an adductor plasty. Patients were evaluated with a minimum follow up of two years. MATERIAL AND METHODS: Forty five females and two males were operated with an average age of fifty years. The pre operative metatarsus varus was of 15 degrees 8. Mean alignment of metatarsal bar was 31 degrees 4. The cuneo metatarsal joint was twenty two times spheric and twenty eight times plane. The average metatarso-phalangeal great toe valgus was 39 degrees 8. RESULTS: They were appreciated with a minimal follow-up of two years, according to the 3 Groulier's criteria: correction of deformation, statics troubles, functional activity. The metatarsus varus improved with an average of 10 degrees 4, as well as the alignment of the metatarsal bar (25 degrees). The post operative average phalangeal valgus was 22 degrees 7. These results were statistically significant. Cuneo-metatarsal joint type did not influenced final result. Articular joint line was normal in 64% of cases. Global result was excellent or good in 70%, passable in 22%, and bad in 8% of cases. DISCUSSION: Scarf Osteotomy of the first metatarsal allows complete correction of metatarsus varus. The surgical approach can be proposed at every age. There are no vascular trouble or arthrosis worsening. It must be completed with a phalangeal varisation or shortening osteotomy and adductor plasty. PMID- 10457557 TI - [Evaluation of the Scarf osteotomy in hallux valgus related to distal metatarsal articular angle: a prospective study of 79 operated cases]. AB - PURPOSE OF THE STUDY: The hallux valgus surgery concerns many operative techniques. The purpose of this report was to evaluate the radiographic results of the Scarf osteotomy related to Distal Metatarsal Articular Angle (DMAA). MATERIAL AND METHODS: The Scarf osteotomy was performed on 79 feet with an hallux valgus deformity (75 females and 4 males from 16 to 82 years). Pre and postoperative weight-bearing X-rays were performed; We measured the Distal Metatarsal Articular Angle (DMAA), the intermetatarsal angle (M1-M2, M1-M5), the metatarsophalangeal angle, the obliquity of the metatarsocuneiform joint and the length of the first and second metatarsal. RESULTS: All parameters were statistically significantly improved by the surgery. However correction on the DMAA was not sufficient (from 14.6 degrees to 12.6 degrees although normal value is 6 degrees). We found a correlation between the metatarsophalangeal angle and the DMAA. DISCUSSION: The preoperative measures showed that the DMAA value increases with the value of the metatarsophalangeal angle; it is insufficiently decreased in comparison to the normal value. It has to be strongly taken in consideration in the preoperative evaluation. CONCLUSION: The preoperative DMAA value is the most important factor in the hallux valgus evaluation. It determines if Scarf or other operative technique are indicated. Its normalisation may provide less recurrences. PMID- 10457558 TI - [Somatosensory evoked potentials during surgery of scoliosis: significance of epidural recording]. AB - PURPOSE OF THE STUDY: To evaluate the usefulness of the epidural recording in scoliosis surgery in children or young adults. MATERIAL AND METHOD: We used somatosensory-evoked potentials in 60 children or young adults in surgical treatment of scoliosis. Somatosensory-evoked potentials were elicited and recorded using a standard method recording of cortical P40 and subcortical P27, simultaneously with the monitoring of spinal cord function using epidural electrode to record the ascending somatosensory volley (by tibial nerve stimulation) at a high thoracic level. RESULTS: The use of subcortical and epidural recording sites for the somatosensory-evoked potentials indicated that a reliable response could be obtained in 96 p. 100 of the patients. Cortical somatosensory-evoked demonstrated a 48 p. 100 false positive rate. DISCUSSION: Epidural recording in scoliosis surgery is safe and reliable. Combined subcortical and epidural recordings are especially useful in patients with neuromuscular scoliosis or with cerebral palsy that may not have reliable cortical potentials. PMID- 10457559 TI - [Isolated radiocarpal luxation. A case report and general review of the literature]. AB - PURPOSE OF THE STUDY: We describe a case of an isolated radiocarpal dislocation and review the literature. CASE REPORT: A young 29-year-old male, suffering from a wrist injury after a motor cycle accident, was diagnosed as dorsal radiocarpal joint dislocation with distal radioulnar dislocation and no fracture, close reduction was immediately performed followed by plaster cast immobilisation. After five months satisfactory stability and motion were obtained without signs of carpal instability. DISCUSSION: This pathology is exceptional with less than 15 published cases. Dislocation is due to wrist hyperextension with ulnar motion but without, probably, intracarpal supination. After immediate treatment, this lesion appeared to have a good prognosis without post-traumatic carpal instability, when compared to the poorer result of perilunar dislocation. PMID- 10457560 TI - [Iatrogenic cervical fracture after femoral diaphyseal nailing]. AB - Three fractures of the femoral neck after intramedullary nailing for diaphyseal fracture are reported. In two cases, the diagnosis was established after nailing but the iatrogenic etiology may be not sure because of an imperfect primary radiological exploration. The localisation of the entry point and the rigidity of Grosse-Kempf's nail could be a contributing factor of this exceptional complication. PMID- 10457561 TI - [Technical aspects of arthroscopic arthrolysis after total knee replacement]. AB - Arthroscopic arthrolysis is a reliable technique for the treatment of knee stiffness due to arthrofibrosis following ligament replacement or following the treatment of knee fracture. However, its use is uncommon for this indication in total knee arthroplasty (TKA). In this study, we questioned whether or not arthroscopy is a reliable technique for treatment of knee stiffness following TKA, due to arthrofibrosis. MATERIALS AND METHODS: Four men mean aged 54 (38-70) underwent knee arthroscopy for a history of arthrofibrosis following TKA. In two cases the arthrofibrosis had appeared after a primary TKA done for limited range of motion and in two other cases had followed a revision arthroplasty. The mean knee flexion before the TKA was 80 degrees (40-110) and it was 105 degrees (100 120) after performing the TKA. The knees were not mobilized under anesthesia though the mean flexion was 75 degrees (60-80) 15 days after the TKA. The indication for arthroscopy was a painless limited range of motion of the knee. The arthroscopy was performed 6 months (2.5-12) after the TKA and at this time the flexion was limited to 65 degrees (60-80). The extension was limited in 2 cases to 10 degrees and 30 degrees. Patients were evaluated an average of 20 months (8-36) after the arthroscopy. With 2 peripatellar portals we sectioned the adhesions in the suprapatellar pouch, the 2 retinaculars and the adhesive bands in the 2 gutters. Two anterior additional portals were used in case of extension lag. A suction drain was placed and the portals were sutured. A continuous passive motion machine was started in the recovery room. RESULTS: The mean operating time for arthroscopic arthrolysis was 38 minutes (30-60). The mean knee flexion was 116 degrees (100-130) at the end of arthroscopy and was 93 degrees (75-110) at the last review. The mean flexion improved by 31 degrees (15-50). The mean flexion improved by 45% (25-83). The 2 extension lags decreased respectively from 30 degrees to 10 degrees and from 10 degrees to 0 degree. For these 2 patients the increase in range of motion was 70 degrees and 40 degrees respectively. The average amount of bleeding was 200 ml (86-520). There were no complications. DISCUSSION: Few surgeons are experienced in arthroscopy for knee stiffness after TKA. Our results are similar to those reported by most authors. Regarding the technique, the section of the two retinaculars is necessary for the mobility of the patellar and most of the mobility is gained after the release of the gutters. The use of only two portals avoids damaging the TKA component and decreases the theoretical risk of infection. The major loss of motion after arthrolysis occurred during the first days following the arthroscopy. This is why we recommend using a regional anesthesia for the arthroscopy and during the following days to allow intensive mobilization of the knee. The arthrolysis should be done from 3 to 6 months after the TKA for better results. CONCLUSION: Arthroscopy for the treatment of knee stiffness, due to arthrofibrosis, following TKA is a useful, reliable and safe technique. PMID- 10457562 TI - MRI of the female pelvis. AB - MRI is a proven modality to evaluate the female pelvis. Excellent soft tissue contrast, sensitivity for the detection of fluid, and the multiplanar imaging capabilities of MR allow noninvasive demonstration of normal anatomy and pathological processes. Most female pelvic MRI studies are performed to answer specific questions and must, therefore, follow carefully developed protocols, which are discussed in this article. The principal MRI techniques and strategies outlined in this work include: (1) the evaluation of reproductive dysfunction, anatomic variants; (2) specific obstetrical applications; (3) oncologic evaluation and tumor staging; (4) problem solving (i.e., the characterization of abnormalities detected by ultrasound); and (5) the evaluation of urethral disease. PMID- 10457563 TI - Screening for gynecological malignancy. AB - Treatment for gynecological malignancy depends for its efficacy at least in part on the stage at presentation. Earlier diagnosis would allow the opportunity for more effective and potentially curative treatment. As a consequence, and in common with initiatives for many other cancers, a search for effective methods of screening is a high priority for the detection of early gynecological cancer. Such methods already exist for cervical cancer, and in many countries screening programs are in place to provide such early diagnosis. Patients with endometrial cancer often present symptomatically at stage I and as a consequence the value of screening of asymptomatic patients may be of lesser importance than for other cancers. Ovarian cancer, however, characteristically presents late and is insidious in onset and progress. Transvaginal ultrasound, together with serum tumor markers, may offer the possibility of early diagnosis and modification of therapy with the potential for improved outcome. However, the evidence from the literature is at present confusing, and it is worthwhile to review the current status of research data to evaluate the place of screening procedures for ovarian and other gynecological malignancies. PMID- 10457564 TI - Pelvic Doppler. AB - This article reviews gray-scale and Doppler evaluation of the adexae and endometrium. Color Doppler is useful in distinguishing between cystic and solid masses. However, spectral Doppler is of limited utility in improving the gray scale morphologic impression of a mass being either benign or malignant. PMID- 10457565 TI - Sonohysterography: technique, endometrial findings, and clinical applications. AB - Transvaginal ultrasound-guided saline infusion sonohysterography (SHG) is a relatively new technique for evaluation of the uterine cavity. As new studies declare the clinical usefulness of this technique, SHG may quickly become part of the routine ultrasound evaluation of the female pelvis. In this article, the SHG procedure is described and normal findings are discussed. Common endometrial pathological findings such as atrophy, polyps, fibroids, hyperplasia, and carcinoma are reviewed. The newly touted roles of SHG for screening patients on long-term tamoxifen therapy and for evaluating patients with infertility are introduced. Finally, we present a practical ultrasound-based flow chart for the workup of menopausal and perimenopausal patients with abnormal uterine bleeding. PMID- 10457566 TI - Sonographic assessment of endometrial disorders. AB - This overview describes the clinical applications of transvaginal sonography in the evaluation of women with suspected endometrial disorders. The role of sonohysterography is also presented as an additional test when evaluating women for possible endometrial disorders. PMID- 10457567 TI - Uterine adenomyosis: current concepts in imaging. AB - This article reviews the clinical findings, histopathological features, and treatment options of uterine adenomyosis and relates them to the role of the radiologist in evaluating patients with this disease. Knowledge of the histopathology of this disease is essential to understanding the range of imaging findings encountered in patients with uterine adenomyosis. In addition, the imaging features and accuracy of noninvasive diagnostic modalities that have proven useful in diagnosing adenomyosis will be discussed. Correctly diagnosing the presence of adenomyosis, and determining its extent, is crucial in planning the appropriate type of therapy. PMID- 10457568 TI - The effect of hydrostatic pressure on intervertebral disc metabolism. AB - STUDY DESIGN: By the use of pressure vessels, hydrostatic pressure was applied to intervertebral disc cells cultured in an alginate. OBJECTIVE: To test the hypothesis that hydrostatic pressure directly affects the synthesis of collagen and proteoglycan by the intervertebral disc cells. SUMMARY OF BACKGROUND DATA: The influence of compression (both hydrostatic and mechanical) on chondrocyte metabolism was examined in a number of earlier studies. However, in most of these studies, articular cartilage, not intervertebral disc, was used, and in none of these was hydrostatic pressure applied to intervertebral disc cells cultured in alginate. METHODS: Fresh cells were harvested from the lumbar intervertebral discs of dogs. Before their suspension in an alginate gel system, the cells were plated and expanded until they reached confluence. Then, by use of the alginate gel system, the cells were exposed (for up to 9 days) to specific values of hydrostatic pressure inside two stainless steel pressure vessels. One vessel was kept at 1 MPa and the other at atmospheric pressure. The effects of 1 MPa were compared against atmospheric pressure by measuring the incorporation of [3H] proline and [35S]-sulfate into collagen and proteoglycans, respectively, for the anulus cells and nucleus cells separately, and by determining whether this incorporation was reflected by changes in the levels of mRNA for aggrecan and Types I and II collagen. RESULTS: Comparisons with atmospheric pressure yielded the following findings: 1) In the incorporation studies, the nucleus and anulus cells exhibited a differential response to a hydrostatic pressure of 1 MPa. Collagen and proteoglycan syntheses were stimulated in the nucleus cells and inhibited in the anulus cells. 2) There was no significant increase in cell proliferation, as measured by DNA content, at 1 MPa for either the anulus or nucleus cells. 3) The mRNA levels of collagen (Col 1A1 and Col 2A1) and aggrecan increased at 1 MPa in both the nucleus and anulus cells. CONCLUSIONS: Hydrostatic pressure directly affects the synthesis of collagen and proteoglycan by the intervertebral disc cells. PMID- 10457569 TI - The role of mast cells in disc herniation inflammation. AB - STUDY DESIGN: A study of herniated lumbar disc tissue samples and control disc material to determine the presence of mast cells in disc herniations. OBJECTIVES: To analyze whether mast cells have any involvement in disc herniation pathophysiology and lumbar pain, because mast cells may have an important role in acute and chronic inflammatory responses. SUMMARY OF BACKGROUND DATA: Studies of inflammatory cells, biochemical mediators of inflammation, and tissue degrading enzymes have suggested that these factors may be involved--and perhaps play an important role--in the pathophysiology of lumbar pain and radiculopathy. Mast cells are known to play an important role in acute and chronic inflammatory responses. It was therefore of interest to clarify their possible role in intervertebral disc herniation inflammation. METHODS: Fifty herniated lumbar discs from 50 patients who had undergone disc surgery and three normal control discs were obtained. Sections from every disc then were examined histologically and immunocytochemically for mast cells by using monoclonal antibodies to either of two types of specific proteases of mast cells, tryptase and chymase. RESULTS: By none of the methods could any mast cells be observed in any of the control disc samples. With toluidine blue staining, mast cells were observed in 9 of 50 (18%) of discs. Mast cells immunoreactive to either tryptase or chymase were observed in 10 of 50 disc samples (20%) and immunoreactive for tryptase and chymase simultaneously in 4 of 50 disc samples (8%). However, the majority of the samples studied (80%) demonstrated immunoreactivity to neither tryptase nor chymase. Among the samples studied were five disc protrusions that totally lacked mast cells. CONCLUSIONS: A minority of disc herniations exhibited mast cells, as verified by toluidine blue staining and immunocytochemistry. The results may suggest a role of mast cells in intervertebral disc herniation inflammation, but only in a subset of these cases. Massive infiltration by mast cells never was observed. PMID- 10457570 TI - The use of an injectable, biodegradable calcium phosphate bone substitute for the prophylactic augmentation of osteoporotic vertebrae and the management of vertebral compression fractures. AB - STUDY DESIGN: A biomechanical study comparing two materials for augmentation of osteoporotic vertebral bodies and vertebral bodies after compression fracture. OBJECTIVES: To compare an injected, biodegradable calcium phosphate bone substitute with injected polymethylmethacrylate bone cement for strengthening osteoporotic vertebral bodies and improving the integrity of vertebral compression fractures. SUMMARY OF BACKGROUND DATA: Injection of polymethylmethacrylate bone cement into fractured vertebral bodies has been used clinically. However, there is concern about thermal damage to the neural elements during polymerization of the polymethylmethacrylate bone cement as well as its negative effects on bone remodeling. Biodegradable calcium phosphate bone substitutes have been studied for enhancement of fixation in fractured vertebrae. METHODS: Forty fresh osteoporotic thoracolumbar vertebrae were used for two separate parts of this study: 1) injection into osteoporotic vertebrae: intact control (n = 8), calcium phosphate (n = 8), and polymethylmethacrylate bone cement (n = 8) groups. Each specimen then was loaded in anterior compression until failure; 2) injection into postfractured vertebrae: calcium phosphate (n = 8) and polymethylmethacrylate bone cement (n = 8) groups. Before and after injection, the specimens were radiographed in the lateral projection to determine changes in vertebral body height and then loaded to failure in anterior bending. RESULTS: For intact osteoporotic vertebrae, the average fracture strength was 527 +/- 43 N (stiffness, 84 +/- 11 N/mm), 1063 +/- 127 N (stiffness, 157 +/- 21 N/mm) for the group injected with calcium phosphate, and 1036 +/- 100 N (stiffness, 156 +/- 8 N/mm) for the group injected with polymethylmethacrylate bone cement. The fracture strength and stiffness in the calcium phosphate bone substitute group and those in the polymethylmethacrylate bone cement group were similar and significantly stronger than those in intact control group (P < 0.05). For the compression fracture study, anterior vertebral height was increased 58.5 +/- 4.6% in the group injected with calcium phosphate and 58.0 +/- 6.5% in the group injected with polymethylmethacrylate bone cement as compared with preinjection fracture heights. No significant difference between the two groups was found in anterior vertebral height, fracture strength, or stiffness. CONCLUSION: This study demonstrated that the injection of a biodegradable calcium phosphate bone substitute to strengthen osteoporotic vertebral bodies or improve vertebral compression fractures might provide an alternative to the use of polymethylmethacrylate bone cement. PMID- 10457571 TI - Effect of decompression enlargement laminoplasty for posterior shifting of the spinal cord. AB - STUDY DESIGN: A study to measure the shifts of the spinal cords and the effects of decompression laminoplasty in 65 patients with cervical lesions who underwent computed tomographic myelography before and after laminoplasty. OBJECTIVES: To investigate limitations of the spinal cord posterior shift after laminoplasty and to clarify the optimal decompression areas to obtain effective posterior shifting. SUMMARY OF BACKGROUND: Although several types of laminoplasty have been performed, all procedures share the common purpose of posterior decompression. No previous studies have examined the limitations of posterior decompression or the optimal decompression range. METHODS: The distance from the posterior edge of each vertebral body or disc level to the posterior edge of the spinal cord was measured by computed tomographic myelography. After the posterior shift was determined by calculating the difference between pre- and postsurgical distances, the relations between posterior shift and neck alignment, clinical results, and the areas of decompression were analyzed. RESULTS: The spinal cord shift ranged from a maximum of 6.6 mm to a minimum of 0 mm. Clinically, spinal cord shifts greater than 3 mm were associated with good clinical outcomes. Upward or downward advanced laminoplasty was related to larger spinal cord shifts at the upper or lower cervical spine. CONCLUSIONS: A mean spinal cord shift of > 3 mm was associated with good clinical outcomes after laminoplasty. In cases with compressive lesions at the upper or lower cervical spine, extension of decompression one level above or one level below likely results in a greater posterior spinal cord shift at these lesions. PMID- 10457572 TI - Long-term clinical and magnetic resonance imaging follow-up assessment of patients with lumbar spinal stenosis after laminectomy. AB - STUDY DESIGN: A cross-sectional retrospective study to observe the correlation between postoperation findings shown on magnetic resonance imaging and clinical observations of 56 patients 10 years after laminectomy for lumbar spinal stenosis. OBJECTIVE: To evaluate the relation between postoperation findings on magnetic resonance imaging and surgical outcome in patients surgically treated for lumbar spinal stenosis. SUMMARY OF BACKGROUND DATA: Surgical management of lumbar spinal stenosis is based on the compression seen in radiologic imaging of neurovascular structures in the vertebral canal, but the success of surgical decompression and its correlation with clinical observations very seldom have been monitored by postoperation radiologic imaging. METHODS: In this study, 56 patients surgically treated for lumbar spinal stenosis were re-examined clinically by use of the Oswestry disability questionnaire. Their walking capacity was evaluated by the treadmill test. Severity of pain before and after the treadmill test was investigated using a visual analog scale. Patients' perception of improvement measured as the change in their condition during the preceding 5 years was elicited by a questionnaire. On the basis of the stenotic findings on magnetic resonance imaging, the patients were classified into no stenosis (NoSten, n = 15) and stenosis (Sten, n = 41) groups, and a summative degenerative scale also was constructed with the findings categorized as follows: disc degeneration, disc herniation, facet joint arthrosis, and degenerative spondylolisthesis. RESULTS: Whereas the patients' perception of improvement correlated very strongly with the Oswestry score and walking capacity, there was no statistical difference between the NoSten and Sten groups in the Oswestry score, walking capacity, perception of improvement, or severity of pain. The effect of the summative degenerative scale on the patients' walking capacity was 13 times greater than the effect of the minimum area of the dural sac. CONCLUSIONS: Patients' perception of improvement had a much stronger correlation with long-term surgical outcome than structural findings seen on postoperation magnetic resonance imaging. Moreover, degenerative findings had a greater effect on patients' walking capacity than stenotic findings. PMID- 10457573 TI - Luque trolley and convex epiphysiodesis in the management of infantile and juvenile idiopathic scoliosis. AB - STUDY DESIGN: Retrospective analysis of 5-year follow-up data from patients instrumented with Luque trolley with or without convex epiphysiodesis for management of progressive infantile and juvenile idiopathic scoliosis. OBJECTIVE: To assess results, establish predictors of outcome, and suggest more effective surgical interventions. SUMMARY OF BACKGROUND DATA: Initial results have been reported. There are no long-term follow-up studies. METHODS: Luque trolley instrumentation was used in eight patients with idiopathic scoliosis between 1983 and 1984. Luque trolley with convex epiphysiodesis was used in 18 patients between 1984 and 1990. RESULTS: Changes in Cobb angle from 8-week to 5-year follow-up are as follows. For Luque trolley alone, Cobb angle worsened for all patients. For progressive infantile scoliosis managed with Luque trolley and convex epiphysiodesis, Cobb angle worsened in seven, remained unchanged in four, and improved in two patients. Mean age at operation was 3.1 years (range, 1.5-7.4 years), and instrumented spinal growth was 32% of expected growth. Preoperation Cobb angle was 65 degrees (range, 40-95 degrees). Cobb angle at 5-year follow-up was 32 degrees (range, 0-86 degrees), which is predicted by preoperation apical concave rib-spinal angle (P = 0.002) and upper end vertebral tilt (P = 0.04). For juvenile idiopathic scoliosis managed with Luque trolley and convex epiphysiodesis, Cobb angle worsened in three patients and improved in one. CONCLUSIONS: Luque trolley instrumentation alone does not prevent curve progression. Additional convex epiphysiodesis results in curve resolution in some patients, which suggests a growth effect. Both spine and rib factors predict Cobb angle at 5-year follow-up. PMID- 10457574 TI - Thoracic discography in healthy individuals. A controlled prospective study of magnetic resonance imaging and discography in asymptomatic and symptomatic individuals. AB - STUDY DESIGN: A prospective case-control investigation. OBJECTIVES: To determine the responses to thoracic discography of asymptomatic individuals. SUMMARY OF BACKGROUND DATA: Literature regarding lumbar and cervical discography reveals that even morphologically abnormal discs often are not painful, whereas painful discs typically exhibit anular or endplate disruption. METHODS: Ten adult lifelong asymptomatic volunteers, ages 23 to 45 years, underwent magnetic resonance imaging of the thoracic spine, followed by four-level discography. Provocative responses were graded on a scale of 0 (no sensation) to 10 (extreme pain or pressure), and filmed discs were graded using a modified Dallas scheme. Concomitantly, 10 nonlitigious adults (6 men and 4 women, ages 31 to 55 years) experiencing chronic thoracic pain were similarly studied as a control group. RESULTS: The mean pain response in the asymptomatic volunteers was 2.4/10. Three discs were intensely painful (scores of 7/10, 8/10, 10/10), with all three exhibiting prominent endplate irregularities and anular tears typical of thoracolumbar Scheuermann's disease. On discography, 27 of 40 discs were abnormal, with endplate irregularities, anular tears, and/or herniations. Ten discs read as normal on magnetic resonance imaging showed anular pathology on discography. In the group with chronic thoracic pain, the average pain response was 6.3/10 (P < 0.05). Of the 48 discs studied, 24 were concordantly painful, with a pain response of 8.5/10 (P < 0.05); 17 had nonconcordant pain/pressure, with an average pain of 4.8/10 (P < 0.05); and 5 had no response. On magnetic resonance imaging 21 of the 48 discs appeared normal. However, on discography, only 10 were judged as normal. CONCLUSIONS: On discography, thoracic discs with prominent Schmorl's nodes may be intensely painful, even in lifelong asymptomatic individuals, but the pain is unfamiliar or nonconcordant. Thoracic discography may-demonstrate disc pathology not seen on magnetic resonance imaging. PMID- 10457575 TI - Short-term physical risk factors for new episodes of low back pain. Prospective evidence from the South Manchester Back Pain Study. AB - STUDY DESIGN: A prospective population-based cohort study performed in South Manchester, United Kingdom. OBJECTIVES: To determine whether nonoccupational physical activity and indicators of physical stress on the spine predict low back pain in the short term. SUMMARY OF BACKGROUND DATA: There is evidence that physical activity outside the workplace helps to protect against low back pain in the long term. However, such activity may injure or stress the spine in the short term. METHODS: A baseline survey questionnaire identified 2715 adults, aged 18-75 years, with no low back pain at the time of the survey. Information on potential predictors of low back pain also was obtained. New episodes of back pain were identified during the subsequent year. RESULTS: A new low back pain episode occurred in 34% of men and 37% of women. Poor general health at baseline was the strongest predictor of a new episode of pain (men: relative risk (RR) 1.5, 95% confidence intervals (CI) 0.8, 2.7; women: RR 2.2, 95% CI 1.2, 4.0). High weight was associated with subsequent low back pain in women (RR 1.4; 95% CI 1.0, 2.0), but neither height nor weight predicted low back pain in men. A self-rated low level of physical activity was not consistently linked with subsequent low back pain, nor were specific nonoccupational physical activities, apart from home improvement work in men and regular sports in women. CONCLUSION: Although some specific activities may be hazardous to the back, physical activity outside the workplace does not increase the short-term risk of low back pain overall. Leisure time physical activity is not a hazard to the back, whereas poor physical health in both genders and heavier weight in women do increase the risk of new low back pain episodes in the short term. PMID- 10457576 TI - The influence of previous low back trouble, general health, and working conditions on future sick-listing because of low back trouble. A 15-year follow up study of risk indicators for self-reported sick-listing caused by low back trouble. AB - STUDY DESIGN: A 15-year follow-up study. OBJECTIVE: To find risk indicators for self-reported sick-listing because of low back trouble and to evaluate which variables were the most important indicators of work incapacity resulting from low back trouble during the follow-up period of 15 years. SUMMARY OF BACKGROUND DATA: The initial data were obtained from a health survey conducted in a general population from the Municipality of Glostrup, Denmark. The follow-up data included information from the Central Person Register, the Early Retirement Pension Register, and a postal questionnaire regarding self-reported sick-listing because of low back trouble. METHODS: An epidemiologic study, in which logistic regression analyses were used for evaluation of the data. The model used consisted of the variable in question, age, gender, and previous experience of low back trouble, along with interactions. RESULTS: It was found that 22 of 37 variables were risk indicators for later self-reported sick-listing because of low back trouble during the preceding year or the 7 years before the date of follow-up evaluation. In analyzing the most significant variables simultaneously, it was found that information from the initial investigation about sick-listing in general during the previous 10 years, sciatic pain, use of analgesics for low back trouble, previous sick-listing because of low back trouble, and occupation were the most important risk indicators for self-reported work incapacity resulting from low back trouble during the follow-up period of 15 years. CONCLUSION: Findings showed that the strongest prognostic indicators of later sick-listing because of low back trouble involve information from the person about previous sick-listing behavior in general and previous experience of low back trouble episodes, especially if these had been accompanied by sciatic pain, use of analgesics, or previous low back trouble sick-listing. PMID- 10457577 TI - Meta-analysis of normative cervical motion. AB - STUDY DESIGN: Meta-analysis of normative cervical range of motion literature performed by applying summary statistics to range of motion and reliability values reported among studies. OBJECTIVES: To identify reliable and valid methods for measuring active and passive cervical range of motion and to estimate normative values. SUMMARY OF BACKGROUND DATA: Range of motion studies use a variety of measuring instruments and statistical analyses, making it difficult to select the most suitable instruments, procedures, and normative values for clinical application. Reviews of the literature, being limited in scope, have not quantitatively synthesized the literature. METHODS: Range of motion and reliability data were grouped by technology and types of motion, then summarized by deriving means and variabilities. Clinical validity was assessed by examining discrepancies, variabilities, and correlations. Change in range of motion as a function of age was determined by comparing range of motion ratios (fourth:third and seventh:third decades). RESULTS: Nine technologies were identified. Overall, passive motion was greater than active motion, and range of motion decreased as age increased, with women exhibiting greater range of motion than men. Variations within each technology were as large as or larger than those between technologies, indicating that clinical procedures are as important as the accuracy and precision of the technology itself. Reliability has not been adequately tested for the majority of technologies. CONCLUSIONS: Clinical procedures appear to be as important as accuracy and precision in determining the reported range of motion values. Further research is needed to establish a gold standard for normative values and to identify an instrument that is reliable for all motions. PMID- 10457578 TI - Randomized trial comparing interferential therapy with motorized lumbar traction and massage in the management of low back pain in a primary care setting. AB - STUDY DESIGN: A randomized trial designed to compare interferential therapy with motorized lumbar traction and massage management for low back pain in a primary care setting. OBJECTIVE: To measure and compare the outcome of interferential therapy and management by motorized lumbar traction and massage. SUMMARY OF BACKGROUND DATA: Management of low back pain by interferential therapy and motorized lumbar traction and massage is common in Germany. No reports of previous randomized trials for the outcome from interferential therapy were found. METHODS: Consenting patients were randomly assigned into one of two groups. A pretreatment interview was performed by the patient using a computer based questionnaire. It also incorporated the Oswestry Disability Index and a pain visual analog scale. Management consisted of six sessions over a 2- to 3 week period. Oswestry Disability Indexes and pain visual analog scale scores also were obtained immediately after and at 3 months after treatment. RESULTS: A total of 152 patients were recruited. The two treatment groups had similar demographic and clinical baseline characteristics. The mean Oswestry Disability Index before treatment was 30 for both groups (n = 147). After treatment, this had dropped to 25, and, at 3 months, were 21 (interferential therapy) and 22 (motorized lumbar traction and massage). The mean pain visual analog scale score before treatment was 50 (interferential therapy) and 51 (motorized lumbar traction and massage). This had dropped, respectively, to 46 and 44 after treatment and to 42 and 39 at 3 months. CONCLUSIONS: This study shows a progressive fall in Oswestry Disability Index and pain visual analog scale scores in patients with low back pain treated with either-interferential therapy or motorized lumbar traction and massage. There was no difference in the improvement between the two groups at the end of treatment. Although there is evidence from several trials that traction alone is ineffective in the management of low back pain, this study could not exclude some effect from the concomitant massage. PMID- 10457579 TI - The effect of a Mensendieck exercise program as secondary prophylaxis for recurrent low back pain. A randomized, controlled trial with 12-month follow-up. AB - STUDY DESIGN: A prospective, randomized, controlled trial with a stratification block design in which a Mensendieck exercise program was compared with the experience of a control group. OBJECTIVE: To evaluate the effect of a Mensendieck program on the incidence of recurrent episodes of low back pain in patients with a history of the condition who currently are working. SUMMARY OF BACKGROUND DATA: One episode of low back pain increases the risk of further episodes of the condition. The Mensendieck approach combines education and exercise. This approach has been used for many years in Scandinavia and the Netherlands. However, the effects on low back pain have not been evaluated previously in a randomized, controlled trial. METHODS: A total of 77 men and women, mean age 39.6 years (range, 21.2-49.8 years), who had finished treatment for a low back pain episode, were stratified according to incidence of low back pain episodes and symptoms of sciatica over the preceding 3 years. The patients were assigned at random to either the Mensendieck program or a control group. The Mensendieck group received 20 group sessions of exercises and ergonomic education in 13 weeks. At 5- and 12-month follow-up examinations, the patients were assessed for recurrence of low back pain, days of sick leave, low back pain, and functional scores. RESULTS: After 12 months, there was a significant reduction in recurrent low back pain episodes in the Mensendieck group compared with the control group (P < 0.05). There was a trend toward fewer days of sick leave because of low back pain in the Mensendieck group, but no significant differences between the groups. There was reduction in pain and improvement in function in both groups, with no significant differences between the groups. CONCLUSIONS: A secondary prophylaxis Mensendieck exercise program of 20 group sessions significantly reduced the incidence of low back pain recurrences in a population with history of the condition. However, there were no differences between the groups with regard to days of sick leave, low back pain, and function. PMID- 10457580 TI - The value of somatosensory- and motor-evoked potentials in predicting and monitoring the effect of therapy in spondylotic cervical myelopathy. Prospective randomized study. AB - STUDY DESIGN: A 2-year follow-up prospective randomized electrophysiologic and clinical study of patients with spondylotic cervical myelopathy. OBJECTIVE: To assess the value of somatosensory- and motor-evoked potentials in the evaluation and prediction of the effect of therapy. SUMMARY OF BACKGROUND DATA: Previous studies have yielded conflicting data concerning the correlation between the changes in evoked potential parameters and the clinical postsurgical outcome in spondylotic cervical myelopathy. METHODS: Sixty-one patients with magnetic resonance images suggesting spondylotic cervical cord compression and clinical signs of cervical myelopathy were divided into two groups according to the degree of clinical cervical cord involvement. The 49 patients with mild and moderate spondylotic cervical myelopathy were randomized into groups that underwent either surgical or conservative therapy. Patients were evaluated clinically and by the means of somatosensory- and motor-evoked potentials. RESULTS: The clinical and evoked potential changes showed good correlation on the group level, but poor correlation intraindividually. There were no significant evoked potential and clinical group changes after 6 months and 2 years in the mild myelopathy group treated either surgically and conservatively, whereas patients with severe myelopathy displayed significant improvement in clinical and evoked potential parameters after surgery. In a subgroup of patients, the isolated segmental medullar N13 abnormality could potentially predict favorable postsurgical clinical outcome. CONCLUSIONS: Longitudinal evoked potentials showed limited use for evaluating the results of therapy in an individual patient. They could be useful in the group assessment of therapy results and in labeling a subgroup of patients with potentially favorable postsurgical outcome. PMID- 10457581 TI - Anatomic evaluation of two different techniques for the percutaneous insertion of pedicle screws in the lumbar spine. AB - STUDY DESIGN: An in vitro study in which a human cadaver model was used to examine the accuracy of two different techniques of percutaneous pedicle screw insertion in the lumbar spine. OBJECTIVES: To investigate the in vitro misplacement rate of pedicle screw insertion for two different percutaneous techniques: 1) the well established Magerl technique, and 2) a new modified technique. SUMMARY OF BACKGROUND DATA: Numerous anatomic and biomechanical studies have been conducted to analyze the in vitro and in vivo characteristics of pedicle screw insertion in the lumbar spine via an open approach, whereas there is a lack of experimental data concerning the more sophisticated percutaneous procedure. METHODS: Thirty human specimens from L1 to S1 were separated into two groups (A and B). In group A, the screws were placed in accordance with the technique described by Magerl; in group B, a new modified technique developed by the authors' research group was used. After screw placement, the specimens were dissected, and pedicle violations were noted with respect to the degree and direction of the screw misplacement. RESULTS: The dissection of the specimens showed that of 360 pedicle screws, 37 were misplaced. This finding translates into an overall misplacement rate of 10%. With the Magerl technique, 23 pedicle violations (misplacement rate, 13%) were found; with the modified technique, only 14 dislocated pedicle screws (misplacement rate, 8%) were detected. This difference was not statistically significant (P = 0.118). In both groups, there were significantly more medical pedicle violations than lateral (32 medial and 5 lateral screw dislocations). The degree of the screw misplacements varied between 1.0 and 5.0 mm. The majority of pedicle violations (30 of 37) were minor, i.e., less than 3.0 mm. CONCLUSIONS: This in vitro study showed that the percutaneous technique of pedicle screw insertion in the lumbar spine is a safe and reliable procedure. Compared with the well-established Magerl technique, the new modified technique did not decrease the misplacement rate significantly, although less pedicle violations were found in the upper lumbar spine. PMID- 10457582 TI - Anterior cervical reconstruction using titanium cages with anterior plating. AB - STUDY DESIGN: A preliminary outcome assessment study of titanium cage implants with anterior cervical plating in anterior cervical reconstruction. OBJECTIVES: To evaluate the efficacy and safety of using titanium cage implants and anterior plating in cervical reconstruction. SUMMARY OF BACKGROUND DATA: Anterior decompression and interbody fusion is a widely accepted surgical treatment for patients with cervical spondylosis. Tricortical iliac crest autograft has been the gold standard but is associated with morbidity at the bone graft donor site, whereas allograft fibula is associated with pseudarthrosis. Problems such as pseudarthrosis, graft collapse, and extrusion still persist with the accepted method of harvesting and implanting bone autografts. METHODS: Thirty-four patients were treated by channel corpectomy followed by placement of a titanium cage packed with autogenous bone graft from the vertebral bodies to reconstruct the anterior column. An anterior cervical plate was added in 30 of 34 cases that involved decompression of two or more levels. The follow-up period ranged from 24 to 56 months, with an average follow-up period of 32 months, and included examination and radiography. RESULTS: Six months after surgery, there was radiographic evidence of fusion in 97% of the patients. Eighty-eight percent of the patients (30 of 34) did not experience any complications (neither cage dislodgment nor hardware failure). Four patients had complications that included pseudarthrosis (1), extruded cage (1), cage in kyphosis (1), and radiculopathy (1). CONCLUSIONS: Titanium cages provide immediate strong anterior column support with minimum hardware complications and avoid bone graft-site morbidity. Titanium cages, with concomitant use of anterior plating, offer an effective and safe alternative to bone autografts. PMID- 10457583 TI - Extraspinal bone and soft-tissue tumors as a cause of sciatica. Clinical diagnosis and recommendations: analysis of 32 cases. AB - STUDY DESIGN: Between 1982 and 1997, the authors treated 32 patients with sciatica who subsequently were found to have a tumor along the extraspinal course of the sciatic nerve. SUMMARY OF BACKGROUND DATA: Extraspinal compression of the sciatic nerve by a tumor is a rare cause of sciatica. Signs and symptoms overlap those of the more common causes of sciatica (i.e., herniated disc and spinal stenosis). OBJECTIVE: To characterize the unique clinical presentation of these patients and to formulate guidelines that may lead to early diagnosis. METHODS: All pertinent clinical data and studies were reviewed retrospectively, and standard demographic data were collected for analysis. RESULTS: These patients typically sought treatment for an insidious onset of sciatic pain that was constant, progressive, and unresponsive to change in position or bed rest. The mean time to final diagnosis was 11.9 months (median, 6 months). Seventeen patients were able to locate their pain to a specific point along the extraspinal course of the sciatic pain, and a mass was noted in 13 patients. Eighteen of these tumors were in the pelvis, 10 in the thigh, and 4 in the popliteal fossa and calf. CONCLUSIONS: A high index of clinical suspicion is the key to early diagnosis of bone or soft-tissue tumors as a cause of sciatica; special attention should be given to pain pattern, physical examination of the entire course of the sciatic nerve, and selection of proper imaging studies. Routine anteroposterior plain radiography of the pelvis as part of the initial imaging screening process is recommended. PMID- 10457584 TI - Immunologic privilege of the eye. PMID- 10457585 TI - Immune mechanisms in uveitis. PMID- 10457586 TI - Sympathetic ophthalmia: an autoimmune ocular inflammatory disease. PMID- 10457587 TI - Anterior uveitis: clinical and research perspectives. PMID- 10457588 TI - The immunobiology of Acanthamoeba keratitis. PMID- 10457589 TI - Ocular disease in AIDS. PMID- 10457590 TI - Immunosuppression in uveitis therapy. PMID- 10457591 TI - Gene therapy in the treatment of ocular inflammation. AB - Gene therapy may become a powerful therapeutic modality in the treatment of both ocular inflammatory disease and as a means of preventing rejection following tissue transplantation. By directly introducing into ocular cells genes that encode proteins capable of down-regulating the immune response, gene therapy has potential for both therapy and as a method for studying mechanisms of disease. While marked and rapid advances in the study of gene therapy have been realized, technical questions regarding the appropriate vector or the choice of efficacious immunomodulatory protein still remain. PMID- 10457592 TI - Stochastic fractal behavior in concentration fluctuation and fluorescence correlation spectroscopy. AB - Fluctuations in the concentration of Brownian particles in one and two dimensions, or any reasonable measurement of the concentration such as in fluorescence correlation spectroscopy, is shown to be a stochastic fractal with a long tail. Being singular at omega = 0, the power spectrum of the fluctuation S(omega) approximately omega-1/2 for diffusion in one dimension, approximately log omega in two dimensions, but non-singular in three dimensions. This discovery provides one simple physical mechanism for possible long-memory fractal behavior, and its implications to various biological processes are discussed. PMID- 10457593 TI - Identification of beta-turn and random coil amide III infrared bands for secondary structure estimation of proteins. AB - Fourier transform infrared spectroscopy is increasingly becoming an important method to determine secondary structure of peptides and proteins. Among the spectral regions arising out of coupled and uncoupled stretching and bending modes of amide bonds, amide I and amide III spectral bands have been found to be the most sensitive to the variations in secondary structure folding. Amide I spectral region (1700-1600 cm-1), although most commonly used primarily because of its strong signal, suffers from several limitations, including a strong interference from water vibrational band, relatively unstructured spectral contour, and overlap of revolved bands correspondingly to various secondary structures. In contrast, amide III spectral region (1350-1200 cm-1), albeit relatively weak in signals, does not have the above limitations. Easily resolved and better defined amide III bands are quite suitable for quantitative analysis of protein secondary structure. While amide III region has been successfully used for determination of alpha-helix and beta-sheets (Fu, F.-N., et al. (1994) Appl. Spectrosc. 48, 1432-1441), bands corresponding to beta-turns and random coils have not been identified, so far. In this paper, we describe, for the first time, identification of amide III bands corresponding to beta-turns and random coils by selectively enhancing random coils by treatment with a denaturing reagent, and secondary structure estimation of several proteins by using the band assignments. The assignments of spectral bands were as follows: 1330-1295 cm-1, alpha-helix; 1295-1270 cm-1, beta-turns; 1270-1250 cm-1, random coils; and 1250-1220 cm-1, beta-sheets. The estimations of secondary structural elements by the above assignments correlated quite well with secondary structure estimations from X-ray crystallography data. PMID- 10457594 TI - Temperature and domain size dependence of sickle cell hemoglobin polymer melting in high concentration phosphate buffer. AB - Deoxygenated sickle cell hemoglobin (Hb S) in 1.8 M phosphate buffer, and carbon monoxide (CO) saturated buffer were rapidly mixed using a stopped-flow apparatus. The binding of the CO to the Hb S polymers and the polymer melting was measured by time resolved optical spectroscopy. Polymer melting was associated with decreased turbidity, and CO binding to deoxy-Hb S was monitored by observation of changes in the absorption profile. The reaction temperature was varied from 20 degrees C to 35 degrees C. Polymer domain size at 20 degrees C was also varied. The data for mixtures involving normal adult hemoglobin (Hb A) fit well to a single exponential process whereas it was necessary to include a second process when fitting data involving Hb S. The overall Hb S-CO reaction rate decreased with increasing temperature from 20 degrees C to 35 degrees C, and increased with decreasing domain size. In comparison, Hb A-CO reaction rates increased uniformly with increasing temperature. Two competing reaction channels in the Hb S-CO reaction are proposed, one involving CO binding directly to the polymer and the other involving CO only binding to Hb molecules in the solution phase. The temperature dependence of the contribution of each pathway is discussed. PMID- 10457595 TI - Interaction of type-I collagen with phospholipid monolayer. AB - The effects of type-I collagen on dipalmitoyl phosphatidylcholine (DPPC) and dimyristoyl phosphatidylcholine (DMPC) monolayer films with different compositions were studied using monolayer technique. The addition of collagen in the subphase of different monolayer films induced a considerable shift towards larger area/molecule in the compression-isotherm curves. This is either referred to the insertion of collagen into the monolayer by its hydrophobic residues or to an adsorption process causing a protein layer to be located parallel to the lipid monolayer [1]. The variation of collagen interaction with different lipid compositions was also verified through the penetration-kinetics experiment. Comparing our results to the results of Pajean et al. [2] and Pajean and Herbage [3] on the effect of collagen on the stability of lipid vesicles implies that the collagen induced stability could be explained on the basis of collagen-lipid monolayer interaction. PMID- 10457596 TI - Interaction of the local anaesthetic heptacaine with dipalmitoylphosphatidylcholine liposomes: a densimetric study. AB - Interaction of the local anaesthetic heptacaine, monohydrochloride of [2 (heptyloxy)-phenyl]-2-(1-piperidinyl)-ethyl ester of carbamic acid, with multilamellar dipalmitoylphosphatidylcholine (DPPC) liposomes in aqueous solution with high excess of water has been studied by means of density measurements in the scanning regime in the main phase transition region. The anaesthetic decreased the temperature of main phase transition. The molar partition coefficients of heptacaine between aqueous phase, liquid crystal and gel phases of DPPC have been determined from a combination of phase transition data obtained by densimetry with a DPPC/heptacaine phase diagram published in the literature. The saturation of heptacaine concentration in liposomes has been observed at higher total amount of anaesthetic. The partial specific volume of heptacaine located in DPPC bilayers is slightly lower than in the aqueous phase. PMID- 10457598 TI - Olfactory learning abilities in staggerer mutant mice. AB - Staggerer mutant mice were compared to non-mutant mice in two olfactory learning tasks. It was found that, in spite of a delayed acquisition compared to non mutants, staggerer mice were able to learn an olfactory habituation task. On the other hand, staggerer presented deficits in an associative olfactory task and, contrary to non-mutants, did not learn this task. Perturbations in olfactory bulbs of staggerer mice could explain their olfactory learning deficits. PMID- 10457597 TI - Molecular phylogeny of Elephantidae. Extreme divergence of the extant forest African elephant. AB - A phylogenetic study of the Elephantidae (Proboscidea, Mammalia) is based on the cytochrome b mitochondrial gene: 31 terminals, that is, all known sequences, one non-elephantid proboscidean, the extinct American mastodon, and four outgroups. The data set includes 11 new sequences with the first published sequence of the forest African elephant, L. a. cyclotis. The analyses of extant taxa only and of both extant and extinct taxa show that L. a. cyclotis is highly divergent from L. a. africana. It is as divergent from L. a. africana as Loxodonta is divergent from Elephas. Southern L. a. africana form a clade. The continental subspecies E. m. indicus is paraphyletic with individuals from India and Thailand closer to E. m. maximus (Sri-Lanka). Members of Mammuthus primigenius are more closely related to Loxodonta although they do not form a clade; two specimens of M. primigenius are closer to L. a. africana making the genus Loxodonta paraphyletic. The latter conclusion may be partly due to unequal length of the various polymorphic mammoth sequences. PMID- 10457599 TI - Karyological and dental identification of Microtus limnophilus in a large focus of alveolar echinococcosis (Gansu, China). AB - A study of voles (Arvicolidae, Rodentia) from Gansu (China) designed to identify a potential host of Echinococcus multilocularis, responsible for human alveolar echinococcosis, leads to a general analysis of Microtus limnophilus population karyotypes, M1 of M. oeconomus populations from all of Eurasia and of M. limnophilus of Mongolia. The Microtus of Gansu belonging to the nominal subspecies M. limnophilus limnophilus (2n = 38; NF = 58) differs markedly in size and shape of M1 from the M. limnophilus of Mongolia, which must therefore be considered as a new subspecies M. limnophilus of malygini nov. ssp. (2n = 38; NF = 60) and the M. oeconomus of Mongolia should be ranked as M. oeconomus kharanurensis nov. ssp. (2n = 30; NF = 60). PMID- 10457601 TI - The uroprotection of mesna on cyclophosphamide cystitis in rats. Its consequences on behaviour and brain activities. AB - We studied the uroprotective effect of mesna, at doses of 40-300 mg/kg/i.p., in single or fractioned injections, on the development of cyclophosphamide (CP, 100 mg/kg/i.p.) cystitis in rats. The study concerns the histological, behavioural and nervous aspects of the disease. The specific effects of mesna, when injected alone, have also been considered. The mesna itself does not have specific deleterious effects, except at a dose of 300 mg/kg which provokes a moderate vesical inflammation although without consequence on the animal's behaviour. Mesna offers good protection against CP cystitis for only certain posologies. The uroprotective effects of mesna reach maxima at doses of 40-100 mg/kg and for fractioned injections given over the entire time frame of the urinary toxic release. The uroprotective effects of other posologies are only partial. The nervous activities were studied through the expression of Fos protein. The repetitive intraperitoneal injection of mesna induced a spinal activity and a preferential contralateral activity of the trigemino/reticular areas of the brainstem spinal cord junction--an effect which was reduced in the presence of CP. The prevention of cystitis by mesna was accompanied only by a reduction in spinal Fos activity, the supraspinal activities remaining high and in strict relationship with the vagal afferent activity. In conclusion, the uroprotective effect of mesna, which requires appropriate posologies, has led to the confirmation of the spinal actions of the CP cystitis, probably via the pelvic nerve, but did not allow a clear distinction between the consequences of the systemic (vagal) and local (spinal, pelvic) actions of CP at supraspinal level. PMID- 10457600 TI - Interspecific and progeny relationships in the genus Stylosanthes inferred from chloroplast DNA sequence variation. AB - The chloroplast trnL (UAA) intron and trnL (UAA)-trnF (GAA) intergenic spacer region have been sequenced from 37 samples, 36 of which representing 19 Stylosanthes species and one from the related genus Zornia. The DNA sequences were used to study phylogenetic relationships in the tropical forage legume genus Stylosanthes, by means of parsimony analysis using the heuristic search method of the computer program PAUP. The resulting cladograms divide Stylosanthes into four separate clades. Within the clades, species are poorly resolved owing to low sequence divergence. Small intra-specific chloroplast DNA variation is observed in S. humilis, S. scabra and the species complex S. guianensis. Variation between S. humilis populations is considered to be geographically structured. The overall results agree well with previously established inter-specific relationships and provide evidence for the genetic origin of the alloploid species S. hamata, S. scabra, S. ingrata, S. sympodialis, S. subsericea, S. capitata and S. fruticosa. This understanding of evolutionary relationships in Stylosanthes, in combination with biogeographical concepts provides a way of discerning isolated habitats in Central and South America, which may therefore contribute to strategies of plant collecting. PMID- 10457602 TI - Quantitative determination of indapamide in pharmaceuticals and urine by high performance liquid chromatography with amperometric detection. AB - A high-performance liquid chromatographic method with amperometric detection for the determination of the diuretic indapamide using a muBondapak C18 column is developed. The mobile phase consists of an acetonitrile-water mixture (45:55, 5 mM) in KH2PO4-K2HPO4 (pH 4.0). The compound is monitored at +1200 mV with an amperometric detector equipped with a glassy carbon working electrode. A liquid liquid or solid-liquid extraction is performed prior to chromatographic analysis to avoid the interferences found in urine matrix. Percentages of recovery are 88.3 +/- 5.6 and 82.9 +/- 7.8 for liquid-liquid and solid-liquid extraction, respectively. The developed method has a linear concentration range from 25 to 315 ng/mL with a reproducibility in terms of relative standard deviation of 4% for a concentration level of 0.5 microgram/mL and a quantitation limit of 1 ng/mL. The method is applied to the determination of indapamide in tablets and urine obtained from hypertensive patients after the ingestion of Tertensif (indapamide 2.5 mg). PMID- 10457603 TI - Effect of hyaluronic acid on migration, proliferation and alpha-smooth muscle actin expression by cultured rat and human fibroblasts. AB - Hyaluronic acid represents a major constituent of the extracellular matrix and influences numerous cellular processes. We have studied the effect of this molecule on migration, proliferation and alpha-smooth muscle actin expression by rat and human cultured fibroblasts. Depending on the tissue origin of the fibroblasts, hyaluronic acid induced, inhibited or did not affect fibroblast migration, whereas proliferation was never affected. Alpha-smooth muscle actin expression was inhibited by hyaluronic acid in all types of human and rat fibroblasts studied, but the inhibition was stronger and dose dependent for subcutaneous fibroblasts and for those recovered from Dupuytren's nodules. This property of inhibiting alpha-smooth muscle actin expression makes hyaluronic acid a good candidate to exert antifibrotic activity. PMID- 10457605 TI - Identification and distribution of type VI collagen in tendon fibrocartilages. AB - Structural and compositional variations are marked in tendon fibrocartilages, which appear at the insertion to bone and in areas subjected to compressive plus frictional loading regime. We are interested in characterizing the extracellular matrix adaptations in these areas, in an attempt to understand cellular responses to biomechanics. In this work, we have applied immunocytochemistry and an ATP treatment for the ultrastructural identification of type VI collagen, to tendons subjected to compressive forces in different species. Immunocytochemistry, after testicular hyaluronidase or pepsin digestion, revealed the presence of type VI collagen in tensional and compressive areas of the plantaris longus tendon of the bullfrog, in the deep flexor tendon of dogs and rabbits, in the calcanear tendon and the suprapatela of rats and in the gastrocnemius tendon of chickens. In each tendon, the tensional region showed a weak reaction, restricted to the collagen fiber surface. However, the compression region was especially rich in type VI collagen, which accumulates in the interfibrillar spaces and is concentrated around the fibrochondrocytes. Intense reaction was also found in the paratenon. The ATP treatment not only allowed for the detection of the typical ladder-like aggregates of type VI collagen in the same areas identified by immunocytochemistry, but also demonstrated that type VI collagen forms a microfibrillar network around the fibrochondrocytes. Besides the function of organizing groups of collagen fibrils, type VI collagen seems to assemble the pericellular matrix in tendon fibrocartilages, perhaps through physical interactions with the large proteoglycans that concentrate in the same area. PMID- 10457604 TI - Ultrastructural recognition of dendritic cells in the intimal lesions of aortas of chickens affected with Marek's disease. AB - The present study was undertaken to determine whether dendritic cells contributed to the intimal abnormalities seen in the aortas of chickens infected with Marek's disease. Semithin sections were used to select aortic segments in which the intima was thickened by lymphocytic infiltrates. Ultrathin sections were then examined. Intermingled with lymphocytes within the intimal thickenings were cells with an appearance typical of dendritic cells including long processes and a unique tubulovesicular system. In these cellular processes the tubulovesicular system was hypertrophied with the tubular cisterns characteristically forming concentric cyclical layers. Our observations show that dendritic cells are present in the aortic lesions of Marek's disease and are presumably involved in related immune mechanisms. PMID- 10457606 TI - Ultrastructural pathology of Golgi apparatus of nerve cells in human brain edema associated to brain congenital malformations, tumours and trauma. AB - The alterations induced by ischemia and anoxia upon smooth endoplasmic membranes are studied in 38 patients with congenital malformations, brain tumours and brain trauma. The effects of vasogenic and cytotoxic brain edema are examined in the Golgi apparatus of nerve and endothelial cells. Samples of cortical biopsies were conventionally processed for transmission electron microscopy. Cortical biopsies were performed according to the basic principles of Helsinki declaration. Slices of 2 to 5 mm were immediately fixed in the surgical room in 4% glutaraldehyde-0.1 M sodium phosphate buffer, pH 7.4 at 4 degrees C, and postfixed in 1% osmium tetroxide in similar buffer. The pathological alterations of the Golgi complex were studied in samples with moderate and severe brain edema. Moderate edema was mainly found in congenital malformations and severe edema in brain trauma and tumours. In some severely edematous neurons, observed in hydrocephalus associated to Arnold-Chiari malformation, small vesicular type Golgi complexes and atrophic ones were observed, characterized mainly by partial or total disappearance of stacked Golgi cisternae and presence of congregated vesicular profiles. In brain trauma and tumours the Golgi complex showed enlargement and fragmentation of the stacked cis- medial- and trans-Golgi cisternae and vacuolization of trans-Golgi network. In addition, an increased formation of Golgi and coated vesicles was observed in the cis- and trans-Golgi regions. Most Golgi and clathrin coated vesicles were observed throughout the cytoplasm suggesting an increased vesicular transport. In severe edema the nerve cell plasma membranes appeared fragmented, presumably due to an interference of the protein insertion process into the plasma membrane. In brain trauma, a hypertrophic Golgi complex was observed in some nerve cells and endothelial cells of cortical capillaries, with increased formation of Golgi and coated vesicles. The ischemia and anoxia associated to the vasogenic and cytotoxic brain edema induced enlargement, fragmentation and disappearance of stacked Golgi cisternae. PMID- 10457608 TI - A light microscopy and ultrastructural study of the testes of tortoise Testudo graeca (Testudinidae). AB - Adult males of Testudo graeca were used to preliminarily study the light microscopic morphology and the ultrastructure of the testes. Spermiogenesis has shown the presence of some interspecific variations among Chelonia, while the general features of the testes and spermatocytes are morphologically similar to other reptilians. The male reproductive state observed in the months analysed has shown spermatogenesis recrudescence in spring, a complete germinal series in autumn and testicular regression in winter. The observation of ultrastructural features, characteristic of steroidogenic activity, suggests a synchrony in tubular and interstitial compartments in T. graeca, with little steroidogenic activity in winter and active synthesis in spring and autumn. In conclusion, the results of this histological study suggest a probable asynchrony between the male and female reproductive cycle in this species and show synchrony in the steroidogenic activity of Sertoli and Leydig cells. PMID- 10457607 TI - Numerous huge iron particles appeared around micro blood vessels under intestinal epithelial cells in infant mice fed excess iron with protein. AB - In infant mice (2 weeks old) fed a diet with excess iron for one week, numerous huge iron particles around micro blood vessels under the basement membrane of intestinal epithelial cells were observed by electron microscopy. The frequency of occurrence of these particles (0.24 +/- 0.04 micron diameter) was markedly higher in mice fed a casein-based diet than in mice fed an amino acid-based diet. The quantity of these particles in both groups decreased in proportion to the term of 2 or 3 weeks. Changes in morphological features, such as opening of the intercellular junctions between intestinal epithelial cells, were also observed in these experimental groups. On the other hand, in mice fed the casein-based diet with excess iron, fat globules appeared in the intestinal epithelial tissue (intestinal epithelial cells, interstitial tissue, lympha) and the occurrence of these increased gradually in proportion to the term of feeding. These fat globules were not observed in mice fed the amino acid-based diet with excess iron. These phenomena might be elicited temporally in infant mice fed excess iron together with protein. The mechanisms of fat globule formation remain unclear. PMID- 10457609 TI - Scanning electron microscopy of large intestine (caecum and colon) of rabbit during foetal and post-natal life. AB - The surface pattern of the large intestine mucosa (caecum and colon) of the rabbit was examined using scanning electron microscope (SEM), in stages ranging from 26 days of foetal to 24 days of post-natal life. It was composed of villi in the foetus and in 1-day-old rabbit. By the 16th day of life these villi were replaced by ridges. PMID- 10457610 TI - Amount and distribution of lipofuscin in nerve and satellite cells from spinal ganglia of young adult and aged rabbits. AB - The quantitative aspects of the age-related accumulation of lipofuscin were studied in the rabbit spinal ganglia by stereological methods using the electron microscope. Both neurons and their associated satellite cells were examined. In neurons, the shape and structure of lipofuscin bodies differed in young adults (12-months-old) compared to aged rabbits (79-months-old), whereas substantial changes were not observed in satellite cells. Both in nerve and satellite cells, lipofuscin bodies were scattered singly in young adults, but were often clustered in old animals. Lipofuscin occupied an average of 0.36% of neuronal perikaryal volume in the young adults and 2.55% in the aged rabbits; these percentages are much lower than those observed in the same neurons of other species. In the satellite cells, the corresponding values were 0.29% and 2.02%. In the young adults, the mean size of lipofuscin bodies was significantly greater in neurons than in satellite cells; the size of these bodies increased with age by about 2.7 times in neurons and by about 1.7 times in satellite cells. Consequently, in the old rabbits the mean size of lipofuscin bodies was about 2.2 times greater in neurons than in satellite cells. It has been suggested that lipofuscin bodies are transferred from the neuronal perikaryon to the surrounding satellite cells and then removed via the capillaries. However, the present findings suggest that lipofuscin located in satellite cells represents pigment formed by these cells rather than being transferred from neuronal perikaryon. It would appear that the age-related accumulation of lipofuscin in the rabbit spinal ganglia has little effect on neuronal metabolism. PMID- 10457611 TI - The lysosomal enzymes acid phosphatase and cathepsin D in rats intoxicated with Senna occidentalis seeds. AB - Chronic administration of Senna occidentalis seeds induces an experimental toxic myopathy characterized by skeletal muscle fibers atrophy, decrease in histochemical activity of cytochrome oxidase, and increase of the acid phosphatase activity in muscle fibres at the light microscopic level. The mechanisms that lead to the increase of this lysosomal enzyme activity are not known and could be related to other biochemical disturbs than the mitochondrial function impairment. The main aim of the present study is to localize the acid phosphatase activity using a cytochemical method at transmission electron microscopy level and to quantify cathepsin D in muscle of rats chronically intoxicated with Senna occidentalis seeds by immunoblotting. Acid phosphatase was observed in lysosomes and over profiles of some organelles apparently not involved by lysosomal membrane. In addition immunoblotting demonstrated a decrease in the content of the precursor and of the mature form of cathepsin D in samples of muscles and liver of intoxicated animals. We concluded that there is a selective increase in acid phosphatase activity in muscle--and maybe in other tissues--of animals intoxicated with Senna occidentalis, that can be related to the skeletal muscle atrophy and the intense decrease in weight gain of these animals. Further studies should be performed to establish the mechanisms of selectivity in increase of lysosomal enzymes in different situations and pathological states. PMID- 10457612 TI - Comparative studies on the ultrastructure of the rat lungs after intratumor treatment of Morris hepatoma with rhTNF-alpha and its muteins. AB - The aim of the present study was the comparative analysis of morphological changes found in the lungs of Buffalo rats in the course of Morris hepatoma 5123 after i.t. treatment with recombinant human TNF-alpha (rhTNF-alpha) and its muteins. Modification of the native TNF-alpha molecule and synthesis of mutagenized analogues can prevent undesirable symptoms observed in the case of therapeutic administration of rhTNF-alpha. TNF-alpha has been shown to interact with two distinct membrane receptors (TNF-R): p55R and p75R. Mutagenized mutein V binds selectively with p55R. Mutein VI fails to recognize either TNF-R. The cytokines were applied in a dose of 10 micrograms protein in a cycle of 8 days. The control group consisted of tumor-bearing animals which were given PBS. Ultrastructural examinations were based on transmission electron microscope (TEM). Mutein VI-receiving animals showed enhanced changes of cytotoxic nature. Severe damage to endothelial cells (necrosis inclusive) was observed. Blood vascular lumen showed accumulation of neutrophils and monocytes. Features of enhanced activity of endothelial cells were noted. Focally, within pulmonary alveoli conglomerates of fibrin and fragments of damaged cells were found, with erythrocytes, neutrophils and macrophages in their vicinity. The epithelium of pulmonary alveoli showed signs of considerable damage, including necrosis. The lumen of pulmonary capillaries in rhTNF-alpha-treated animals showed a predominance of eosinophils and monocytic cells. Features of endothelial stimulation were observed, although without a tendency to form microthrombi. Much less pronounced changes both in the lung capillary bed and in the alveolar epithelial cells were noted in the mutein V-given animals. Our findings confirm the possibility of peripheral activation of cells involved in the cytokine induced antitumor response. Mutein V with the smallest effect on the lung tissue rebuilding seems to be a rhTNF-alpha derivative which can delimit the undesirable symptoms in the course of antitumor therapy reduced to i.t. injections. PMID- 10457613 TI - Ultrastructural changes on the epithelial cells of uterine tubes of Wistar rats after chronic ethanol ingestion. AB - The present paper describes the morphological alterations of the epithelial layer of the uterine tubes of rats submitted to experimental chronic alcoholism using anatomical, histological, ultrastructural and morphometric methods. Sixty adult rats (Rattus norvegicus albinus) at the same age (3 months) and with a mean body weight of 228 g were divided into two groups. The control group received solid diet (Purina rat chow) and tap water ad libitum. The alcoholic group received the same solid diet and was allowed to drink only sugar cane brandy dissolved in 30 degrees Gay Lussac (v/v). After periods of 90, 180 and 270 days of treatment animals at normal estrus were anaesthetised with ethyl ether, weighed and sacrificed. Subsequently, the uterine tubes were dissected, weighed and prepared for TEM and SEM methods. The final mean body weights were similar in the control and alcoholic groups. The morphometric analysis showed no difference between control and alcoholic epithelial height. The alcoholic animals showed ultrastructural alterations: intense lipid droplet and lysosomes accumulation, dilated rough endoplasmic reticulum cisternae and vacuolization in both periods of treatment. It was concluded that alcohol acts as a toxin on the epithelial layer of the uterine tubes of rats. PMID- 10457614 TI - Adriamycin induces exocytosis in rat and beige mouse peritoneal mast cells: an ultrastructural, morphometric and biochemical study. AB - The process of exocytosis was studied in rat and beige mouse peritoneal mast cells stimulated by adriamycin (ADR) at 37 degrees C and 22 degrees C. ADR induces a non cytotoxic histamine release that is followed by a significant uptake of the drug. Examination was performed by transmission electron microscopy and, at the same time, histamine release and ADR uptake were measured by spectrofluorimetry. ADR accumulation in mast cells was investigated also by fluorescence microscopy. In rat mast cells stimulated at 37 degrees C, the secretory process developed abruptly and was virtually complete after 30 sec. Electron microscopy showed rapid intracytoplasmic channel formation and extrusion of secretory granules; spectrofluorimetry revealed a massive release of histamine and rapid uptake of ADR. In addition, fluorescence microscopy showed mast cells exhibiting an intense orange-yellow fluorescent signal localized at the secretory granules. At 22 degrees C, rat mast cells showed alteration of the granules, cavity formation by fusion of the perigranular membrane and granule discharge due to fusion of the cavity membrane with the cell membrane. Histamine release and ADR uptake proceeded less quickly than at 37 degrees C. Quantitative analysis of rat mast cell ultrastructure demonstrated that histamine release induced by ADR stimulation was achieved by sequential exocytosis. This process presents both morphological and biochemical affinities with the exocytosis induced by basic secretagogues such as compound 48/80. In beige mouse mast cells the process of exocytosis progressed more slowly and was completed after 20 min at 37 degrees C. By electron microscopy, the cytoplasm presented a rigid structure due to abundance of actin-like fibrils. Granule fusion was an uncommon feature and exocytosis was mostly the result of single granule opening to the cell exterior without extrusion of granule matrices. PMID- 10457615 TI - Characterization of tuberculo-ventral neurons in the dorsal cochlear nucleus of the guinea pig. AB - Tuberculo-ventral neurons in the deep polymorphic layer of the dorsal cochlear nucleus of the guinea pig are immunopositive to glycine, their main neurotransmitter, as revealed by post-embedding immunogold. The initial size of gold particles (10-20 nm) was increased (40-70 nm) by silver intensification for a better and rapid individuation of glycine positive cells at low magnification. The most specific ultrastructural characteristic of tuberculo-ventral neurons is their synaptic profile that shows most flat-pleomorphic axo-somatic boutons. This synaptic profile is different from that of other medium size neurons, such as cartwheel neurons in the superficial layer of the dorsal cochlear nucleus. The synaptic profile of tuberculo-ventral neurons together with other characteristics may explain their activity connected to the neural network responsible for echos suppression or/and sound localization. PMID- 10457616 TI - Studies on mitochondrial pathogenesis of Rett syndrome: ultrastructural data from skin and muscle biopsies and mutational analysis at mtDNA nucleotides 10463 and 2835. AB - In order to verify the pathogenic role of mitochondria in Rett syndrome, the results are reported of skin and muscle biopsies in two RS patients, showing morphological ultrastructural abnormalities in mitochondrial number and size. The investigation on two recently described mtDNA mutations (at nt 10463 and 2835) (Lewis et al., 1995; Tang et al., 1997) gave contrasting results with respect to previously reported data. In particular, the first mutation has been found in seven subjects, five of whom were from two different control groups, while the second mutation has been not detected in any of the 26 patients and controls. Although these results present some signs of a morphological impairment of mitochondria, they do not support the hypothesis that the two mutations may have a primary role in the pathogenesis of the syndrome and indicate the need for further investigations on the role of mtDNA in the pathogenesis of the syndrome. PMID- 10457617 TI - Similar non-random processes maintain diversity in two tropical rainforests. AB - Quadrat-based analysis of two rainforest plots of area 50 ha, one in Panama (Barro Colorado Island, BCI) and the other in Malaysia (Pasoh), shows that in both plots recruitment is in general negatively correlated with both numbers and biomass of adult trees of the same species in the same quadrat. At BCI, this effect is not significantly influenced by treefall gaps. In both plots, recruitment of individual species is negatively correlated with the numbers of trees of all species in the quadrats, but not with overall biomass. These observations suggest, but do not prove, widespread frequency-dependent effects produced by pathogens and seed-predators that act most effectively in quadrats crowded with trees. Within-species correlations of mortality with numbers or biomass are not found in either plot, indicating that most frequency-dependent mortality takes place before the trees reach 1 cm in diameter. Stochastic effects caused by BCI's more rapid tree turnover may contribute to a larger variance in diversity from quadrat to quadrat at BCI, although they are not sufficient to explain why BCI has fewer than half as many tree species as Pasoh. Finally, in both plots quadrats with low diversity show a significant increase in diversity over time, and this increase is stronger at BCI. This process, like the frequency dependence, will tend to maintain diversity over time. In general, these non random forces that should lead to the maintenance of diversity are slightly stronger at BCI, even though the BCI plot is less diverse than the Pasoh plot. PMID- 10457618 TI - Maternal effort and male quality in the bank vole, Clethrionomys glareolus. AB - Parental investment in reproduction is adjusted according to potential benefits in terms of offspring survival and/or mating success. If male quality affects the reproductive success of a female, then females mating with high-quality males should invest more in reproduction. Although the subject has been of general interest, further experimental verification of the hypothesis is needed. We studied whether female bank voles (Clethrionomys glareolus) adjusted their maternal effort according to male quality, measured as mating success. To enable the measurement of maternal effort during nursing separately from male genetic effects the litters were cross-fostered. Further, the genetic background of male quality was examined. Male quality did not correlate with litter size or offspring size at birth. Offspring growth was positively related to food consumption and milk production of mothers. However, these direct measurements of maternal effort were independent of male quality. Male mating success appeared to be significantly heritable indicating that there are genetic benefits. Still, females did not adjust maternal effort according to the genetic quality of their offspring. We suggest that female bank voles gain significant genetic benefits from mating with high-quality males whereas they cannot improve their reproductive success by increasing maternal effort. PMID- 10457620 TI - A nurse by any other name? PMID- 10457619 TI - Paternal kin discrimination in wild baboons. AB - Mammals commonly avoid mating with maternal kin, probably as a result of selection for inbreeding avoidance. Mating with paternal kin should be selected against for the same reason. However, identifying paternal kin may be more difficult than identifying maternal kin in species where the mother mates with more than one male. Selection should nonetheless favour a mechanism of paternal kin recognition that allows the same level of discrimination among paternal as among maternal kin, but the hypothesis that paternal kin avoid each other as mates is largely untested in large mammals such as primates. Here I report that among wild baboons, Papio cynocephalus, paternal siblings exhibited lower levels of affiliative and sexual behaviour during sexual consortships than non-kin, although paternal siblings were not significantly less likely to consort than non kin. I also examined age proximity as a possible social cue of paternal relatedness, because age cohorts are likely to be paternal sibships. Pairs born within two years of each other were less likely to engage in sexual consortships than pairs born at greater intervals, and were less affiliative and sexual when they did consort. Age proximity may thus be an important social cue for paternal relatedness, and phenotype matching based on shared paternal traits may play a role as well. PMID- 10457621 TI - Preparing parents for their child's transfer from the PICU to the pediatric floor. AB - The experimental study described here was conducted to examine a nursing intervention intended to diminish the anxiety level of parents of children being transferred from a pediatric intensive care unit (PICU) to a general pediatric floor. The convenience sample included 50 parents of patients in a PICU at a southeastern U.S. tertiary medical center. The sample was randomly assigned to control and experimental groups. All subjects' anxiety levels were measured using Spielberger's State-Trait Anxiety Inventory (STAI) 24 to 48 hours prior to the child's impending transfer. The experimental group subsequently was given a transfer-preparation letter along with a verbal explanation. Finally, both groups were retested using the STAI immediately prior to the child's transfer. After controlling for trait anxiety, the analysis of covariant results showed significantly lower anxiety levels among the subjects who received the transfer preparation letter than among those who did not, F(1,47) = 18.65, p < .0005. The study concludes that effective transfer preparation can significantly reduce the anxiety experienced by parents who are facing the imminent transfer of their child out of the PICU. PMID- 10457623 TI - Mutuality and preparedness of family caregivers for elderly women after bypass surgery. AB - The purposes of this study were to identify caregivers' feelings of mutuality and reported preparedness for caregiving at hospital discharge and to examine the relationship between these measures and the recovery outcomes of elderly women after coronary artery bypass graft (CABG) surgery. A sample of 49 dyads was followed at discharge, 6 weeks, 3 months, and 6 months. The caregivers' Mutuality Scale (MS) scores and Preparedness for Caregiving (PCS) scores at discharge indicated positive feelings about the relationship with the elderly woman and their preparedness, and both measures were positively correlated with the self report recovery measures at 3 months (p < .05). Nevertheless, at 3 months the MS scores were significantly lower than at discharge (p < .05), indicating stress in the caregiver-patient relationship. The MS and PCS may be useful tools for assessing situations and directing nursing interventions during discharge planning. PMID- 10457622 TI - Hope and anxiety of individual family members of critically ill adults. AB - This correlational study describes levels of hope and states of anxiety of individual family members of critically ill patients and investigates the relationship between hope and state of anxiety. Forty-four family members of critically ill patients participated in the study and revealed a high level of hope and a high level of anxiety within 72 hours of the patient's hospitalization, regardless of personal or situational characteristics. Hope and anxiety were not significantly related in this sample; themes of hope focused on past and present experiences, whereas anxiety centered on death of the loved one. Participants looked to the past, recalling happy times, and to the future, in relation to goals and potential. Family members acknowledged feeling secure, self confident, and steady, and were able to make decisions because they were not confused about the situation. Nurses are challenged to inspire the family's hope and decrease their anxiety. PMID- 10457624 TI - Sitting posture and prevention of pressure ulcers. AB - The aim of this study was to gain insight about the influence of body posture on the pressure at the seat surface and to establish to what extent different seat cushions designed for incontinent patients reduce maximum pressures. Pressures were measured for 56 healthy volunteers in eight postures using four cushions. The posture in which the lowest maximum pressure was measured was the sitting back posture with the lower legs on a rest. If the seat could not be tilted back, the maximum pressure in the upright sitting posture with the feet on the ground was significantly lower than sitting upright with the legs supported on a rest. Sliding down and slouching caused the highest maximum pressure. Regular checking of the posture and using positioning cushions should form part of any pressure ulcer prevention protocol. The four selected cushions each have different pressure-reducing effects. A thick air cushion (Repose) has the lowest maximum pressure and is significantly better than the other cushions at reducing the high pressure when slouching or sliding down. PMID- 10457625 TI - Patient satisfaction with two models of group therapy for people hospitalized with bipolar disorder. AB - The purpose of this study was to (a) compare participants' evaluations of two models of inpatient group therapy for people with bipolar disorder and (b) develop a taxonomy of patient satisfaction with such groups. The sample of 110 people included 59 participants in the self-management groups and 51 participants in the interactional groups. Results indicated that although neither of the models was clearly preferred by participants, the responses were useful in identifying needed changes, and the benefits of providing inpatient diagnostically homogeneous groups for this population were evident. A refined self-management model that incorporates agenda setting in the format, and a taxonomy for use in future research, are presented. PMID- 10457626 TI - Data display techniques. AB - The way data are displayed is as important as the data. The reader's ability to perceive, comprehend, and recall the information contained within the data is affected by how the data are presented. Multiple factors must be considered when designing data displays. Characteristics of the data and the intended use of the data should drive the choice of the display format. Formats for data displays can be words or numbers. Numbers can be displayed as specific values, tables, or graphs. Advantages and limitations of different types of data formats are presented. Techniques for the use of color are also included. PMID- 10457627 TI - Nurses' perceptions of chemical restraint use in long-term care. AB - The purpose of this pilot study is to validate the use of the Perceptions of Restraint Use Questionnaire (PRUQ) in assessing chemical restraint perceptions among nurses working in long-term care. The convenience sample includes 60 licensed nurses working in six long-term care facilities in Illinois. The reliability analysis for a modified version of the PRUQ, based on the research sample chosen, was found to have a Cronbach's coefficient alpha of .9450. Study findings reflect a moderately positive attitude toward chemical restraint use by nurses in long-term care. PMID- 10457628 TI - Effects of head-of-the-bed positioning on the electrocardiogram. PMID- 10457629 TI - How much is enough? PMID- 10457630 TI - Worker exposures during use of a non-chromate aircraft primer (tie coat). PMID- 10457631 TI - Control of smoke from laser/electric surgical procedures. National Institute for Occupational Safety and Health. PMID- 10457632 TI - Acute liver dysfunction among workers exposed to 2,2-dichloro-1,1,1 tryfluoroethane (HCFC-123): a case report. PMID- 10457633 TI - National Academy of Sciences Workshop and Report on the Science of Ergonomics. PMID- 10457634 TI - ELPAT Program. Background and current status. PMID- 10457635 TI - Industrial hygiene recommendations as interventions: a collaborative model within occupational medicine. AB - The authors conducted a cross-sectional telephone survey of patients and employers to evaluate the effectiveness of conducting industrial hygiene work site visits as part of the medical management of clinic patients with suspected occupational disease. Industrial hygiene interventions were intended to keep the index patient from ongoing exposure while simultaneously protecting co-workers from future disease. The demographics of the 76 work sites and patients are summarized. According to the employers, 78 percent had implemented at least one recommended intervention, and 52 percent of the employers had implemented the priority intervention. The factors associated with the employers' implementation of the recommended industrial hygiene controls are presented. Employers were 3.7 times more likely to implement the priority intervention (p = .04) if they believed a worker's illness was work-related. Employers with joint labor management health and safety committees were 3.8 times as likely to implement the priority intervention (p = 0.04). The factors associated with changes in the patients' self-reported disease status are explored and the social and economic implications of this model, are discussed. Patients who reported an improvement in disease status were 5.5 times more likely to have maintained or increased their salary (p = 0.03). Patients were 10.4 times more likely to be working if the employer had implemented any intervention (p = 0.02) and 13.3 times more likely if the priority intervention (p = 0.01) had been adopted. Those patients who left their original employer suffered a median decrease in salary of 35 percent if they found work with a different employer and a 90 percent decrease when they failed to find a new job. PMID- 10457636 TI - Workers' compensation experience of North Carolina residential construction workers, 1986-1994. AB - A total of 31,113 workers' compensation claims among 7,400 North Carolina Homebuilders Association (NCHA) members and their subcontractors for the period 1986-1994 were analyzed to calculate workers' compensation claim incidence density rates. For the 7 years studied, the average rate (cases/200,000 work hours) for all claims was 16.40 and the rate for medical or lost time cases was 10.78. Highest rates for cases involving medical costs or paid lost time by mechanism of injury were observed for being struck by an object (3.1), lifting/movement (1.97), falls from a different level (1.13), striking against an object (0.87), and falls on the same level (0.46). Rates by mechanism of injury were highest for muscle strains (2.34), wounds/punctures (2.33), bruises/contusions (1.24), fractures/dislocations (0.98), and injuries to the eyes (0.81). Among medical cost or lost work time cases, body parts with highest injury rates were back/shoulders (1.99), fingers (1.31), leg/knee (1.00), hand/wrist (1.00), foot/ankle (0.86), and eyes (0.82). Injury rates were found to vary substantially among the residential construction trades. For more serious injuries involving medical costs greater than $2,000 or any lost work time, rates were highest for welders and cutters (28.1), insulators (24.3), roofers (19.4), and carpenters (15.3). The same general trends by trade were observed for cases involving paid lost time except that roofers were highest, with a rate of 9.1, followed by insulators (8.5), welders and cutters (5.8), and carpenters (5.8). Rates of falls from a different level resulting in medical costs or lost work time were highest for roofers (5.54), insulators (3.53), carpenters (2.05), and drywall installers (1.99). Descriptive information for falls from a different level resulting in paid lost time during 1993-1994 (n = 219) were reviewed to better determine the causes and circumstances of injuries. Falls from a roof accounted for 25.4 percent of the cases followed by falls involving scaffolds (23.9%) and ladders (20.6%), and falls from ceiling joists, floor joists, or framing (14.8%). Twenty-six work-related deaths occurred with vehicle accidents (n = 6) being the major known cause of death, followed by falls (n = 3), being struck by an object (n = 3), electric shock (n = 2), and contact with energy or chemicals (n = 2). Consistent with other analyses of workers' compensation data, chronic occupational diseases are not well captured in the workers' compensation claims among home builders; therefore, a companion study has examined mortality patterns among North Carolina construction workers. PMID- 10457637 TI - Laboratory evaluation of a protocol for personal sampling of airborne particles in welding and allied processes. AB - A new European/International Standard (ISOprEN 10882-1) on the sampling of airborne particulates generated during welding and allied processes has been proposed. The use of a number of samplers and sampling procedures is allowable within the defined protocol. The influence of these variables on welding fume exposures measured during welding and grinding of stainless and mild steel using the gas metal arc (GMA) and flux-cored arc (FCA) and GMA welding of aluminium has been examined. Results show that use of any of the samplers will not give significantly different measured exposures. The effect on exposure measurement of placing the samplers on either side of the head was variable; consequently, sampling position cannot be meaningfully defined. All samplers collected significant amounts of grinding dust. Therefore, gravimetric determination of welding fume exposure in atmospheres containing grinding dust will be inaccurate. The use of a new size selective sampler can, to some extent, be used to give a more accurate estimate of exposure. The reliability of fume analysis data of welding consumables has caused concern; and the reason for differences that existed between the material safety data sheet and the analysis of fume samples collected requires further investigation. PMID- 10457638 TI - Continuing the search for WBGT clothing adjustment factors. AB - The original concept for the heat stress limits adopted by the American Conference of Governmental Industrial Hygienists (ACGIH) was that the threshold for heat stress can be marked by environmental conditions (WBGT) that are adjusted for metabolic rate. The underlying data were based on ordinary work clothes. The Physical Agents Committee promoted clothing adjustment factors within the industrial hygiene community through a revision in the TLVs for heat stress in 1990. As approved, there were adjustment factors for three other clothing ensembles. To extend and further understand adjustment factors, adjustments for commercially available clothing ensembles and prototype ensembles have been examined. The fundamental principle of the assignment of an adjustment factor to an ensemble begins with establishing critical environmental conditions in which test subjects were just able to maintain thermal equilibrium. Four or five subjects for each ensemble walked on a tread-mill inside a climatic chamber under controlled conditions of heat stress. During each test, heart rate and core temperature were continuously monitored. After a physiological steady-state was achieved, temperature and humidity were slowly increased to maintain relative humidity at 20 percent, 50 percent, or 70 percent. Metabolic rate was assessed by measuring the rate of oxygen consumption. Reviewing a trial, the critical conditions were noted as the time when thermal regulatory control was lost (called the inflection point and marked by a steady increase in core temperature). The climatic conditions at the inflection point were used to assign a critical WBGT. A WBGT representative of each ensemble was assigned through a weighted average of different protocols. Clothing adjustment factors representing an equivalent increase in WBGT were computed by noting the difference between the representative WBGT of the cotton work clothes and the other clothing ensembles. The results suggested adjustment factors with reference to ordinary work clothes from the least stress at -2.1 to 5.5 degrees C-WBGT. The adjustment factors were reasonably well predicted by a linear regression based on a computed value for total evaporative resistance (r2 = 0.92). PMID- 10457639 TI - Prediction of workplace wet bulb global temperature. AB - The wet bulb globe temperature (WBGT) is the de facto standard to assess environmental contributions to heat stress. A practical problem emerges when the heat stress conditions vary over many locations or during the day. To address this problem, investigators have suggested empirical relationships and thermodynamic models. The purpose of this effort was to examine a thermodynamic model in the laboratory and to predict WBGTs in an aluminum smelter by both the empirical and thermodynamic models. In the laboratory, there was no real difference between the experimental data and the thermodynamic model. In the application to an aluminum smelter, there was a small overall tendency for the predicted values to be greater than the actual values, but there was no practical difference between the models. The empirical model provided a good match with a slight over-prediction by 0.5 degree C with a standard deviation of 3.0 degrees C. For the same data, the thermodynamic model had an average over-prediction of 0.7 degree C with a standard deviation of 2.8 degrees C. Either method of predicting WBGT was effective. The empirical method required less computation and was conceptually simpler. PMID- 10457640 TI - Ozone exposure at a construction site. PMID- 10457641 TI - The ergonomic use of hand tools: guidelines for the practitioner. National Institute for Occupational Safety and Health. PMID- 10457642 TI - NIOSH HHEs on CD-ROM. PMID- 10457643 TI - Control of nitrous oxide in dental operatories. National Institute for Occupational Safety and Health. PMID- 10457644 TI - Beryllium contamination inside vehicles of machine shop workers. AB - Inhalation of beryllium particles causes a chronic, debilitating lung disease- chronic beryllium disease (CBD)--in immunologically sensitized workers. Evidence that very low concentrations of beryllium may initiate this chronic disease is provided by incidences of the illness in family members exposed to beryllium dust from workers' clothes and residents in neighborhoods surrounding beryllium refineries. This article describes the results of a cross-sectional survey to evaluate potential take-home beryllium exposures by measuring surface concentrations on the hands and in vehicles of workers at a precision machine shop where cases of CBD had recently been diagnosed. Many workers did not change out of their work clothes and shoes at the end of their shift, increasing the risk of taking beryllium home to their families. Wipe samples collected from workers' hands and vehicle surfaces were analyzed for beryllium content by inductively coupled argon plasma-atomic emission spectroscopy (ICP-AES). The results ranged widely, from nondetectable to 40 micrograms/ft2 on workers' hands and up to 714 micrograms/ft2 inside their vehicles, demonstrating that many workers carried residual beryllium on their hands and contaminated the inside of their vehicles when leaving work. The highest beryllium concentrations inside the workers' vehicles were found on the drivers' floor (GM = 19 micrograms/ft2, GSD = 4.9), indicating that workers were carrying beryllium on their shoes into their vehicles. A safe level of beryllium contamination on surfaces is not known, but it is prudent to reduce the potential for workers to carry beryllium away from the work site. PMID- 10457645 TI - Transfer of contaminants from surface to hands: experimental assessment of linearity of the exposure process, adherence to the skin, and area exposed during fixed pressure and repeated contact with surfaces contaminated with a powder. AB - Estimation of dermal exposure in the workplace resulting from contact with contaminated surfaces is important in risk assessment. Models have been developed to describe the process of exposure due to transfer, but for major input parameters--that is, contact area surface and adherence--defaults are used. This study examines the effect of one single-hand press contact and repeated contacts with a contaminated glass plate on both skin area exposed and loading of the skin for three volunteers. A fluorescent whitening agent was used to study the process of exposure and to determine the increase of the area exposed as well as the adherence of the compound to the skin surface after 1 to 12 consecutive contacts by a video imaging technique. In addition, loading of the skin after 12 contacts was compared to loading of a cotton glove monitor with similar hand pressures. The results show that after one single-hand contact only 4 to 16 percent of the total surface of the palm of the hand was exposed, whereas after 12 contacts this was increased to about 40 percent. The efficiency of transfer was < or = 2 percent of the contamination of the surface. The adherence to the skin was 1.07 micrograms/cm2 after 12 contacts and tended to increase non-linearly with increase in contacts. Because defaults of adherence for use in exposure models are generally a factor 500 to 5,000 higher, and the area exposed is assumed to be the total surface of the hand, overestimation of dermal exposure due to a single hand-surface contact in workplaces may occur. Therefore, additional studies on multi-contact exposure scenarios are indicated to adjust defaults for hand surface transfer processes. PMID- 10457646 TI - Determination of volatile organohalogen compounds in human urine. AB - A variant of thin layer headspace (TLHS) analysis with autogenous generation of the liquid sorbent was applied for the determination of volatile organohalogen compounds in urine. The analytes were isolated from the matrix at an elevated temperature in a TLHS column. They were then transported by a stream of purified air to a second TLHS column kept at sub-ambient temperature. Water vapor contained in the air condensed in the second column together with the analytes. The effluent from the second column was analyzed by direct aqueous injection gas chromatography with electron capture detection (DAI-GC-ECD). It has been established that the complex urine matrix did not affect the results at a 95% confidence level. Volatile organohalogen compounds were detected in urine of employees of a chemical laboratory where halogenated solvents were used. The urine of people who were not exposed contained insignificant or undetectable levels of these compounds. Urine levels of target analytes were the higher, the longer a person was exposed to them. The results obtained confirm that the urinary system participates in excretion of volatile organohalogen compounds. PMID- 10457647 TI - A pilot study to characterize fine particles in the environment of an automotive machining facility. AB - The main goal of this study was to characterize fine particles (e.g., smaller than about 3 microns) in an automotive machining environment. The Toledo Machining Plant of Chrysler Corporation was selected for this purpose. The effect of local mechanical processes as aerosol sources was a major part of this investigation. To determine the size-dependent mass concentration of particles in the plant, the Micro-Orifice Uniform Deposit Impactor (MOUDI Model 100, MSP Corp., Minneapolis, Minnesota) was used. The MOUDI was placed at central locations in departments with sources inside the plant, so that the obtained information on the size distribution realistically represents the aerosol to which plant workers are exposed. Sampling was conducted over a 4-day period, and during three periods per day, each matching the work shifts. A special effort was made to place the MOUDI at a central location of a department with relatively homogeneous particle sources. The selected sampling sites included welding, grinding, steel machining, and heat treating processes. The average 24-hour mass concentrations of particles smaller than 3.2 microns in aerodynamic diameter were 167.8, 103.9, 201.7, and 112.7 micrograms/m3 for welding, grinding, mild steel, and heat treating processes, respectively. Finally, the mass median diameters of welding, heat treatment, machining, and grinding operations were approximately 0.5, 0.5, 0.6, and 0.8 micron, respectively. PMID- 10457649 TI - Animal study on electromagnetic field biological potency. AB - This recent basic research study used an animal model protocol to assess specific biomarkers of the effect of non-ionising, non-thermal radiation (2450 MHz microwave radiation at 5-15 mW/cm2) on bone marrow, peripheral blood, and bronchoalveolar free cell populations. Of 40 male Wistar rats taken in the study, 20 animals of the experimental group were irradiated for 2 hours a day, 5 days a week, and subsequently killed on days 1, 8, 16, and 30 of the experiment. The remaining 20 rats served as control. All animals were previously intratracheally instilled with biologically inert microspheres to see the influence of irradiation on lung retention kinetics. The cell response to chosen electromagnetic irradiation was followed quantitatively and qualitatively using the standard laboratory methods. The results of peripheral blood cell response suggested a decreasing tendency in total leukocyte count and in relative lymphocyte count in the treated group. A slight increase was also observed in granulocyte count and in the absolute count of peripheral blood erythrocytes over control animals. PMID- 10457648 TI - A new passive sampler for regulated workplace aldehydes. AB - A new solid sorbent passive air sampler for aldehydes had a silicone membrane atop a cylindrical diffusion path of 1.1 cm length and 1.3 cm diameter above a 10 percent (w/w) O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA) Tenax TA pellet. Known vapor concentrations of Occupational Safety and Health Administration (OSHA)-regulated aldehydes near their permissible exposure limits were generated from a syringe pump dynamic air dilution system that was connected to an exposure chamber. The O-oxime derivatives from aldehyde reaction with PFBHA were desorbed with hexane, and quantified by capillary gas chromatography/mass spectrometry (GC/MS) or gas chromatography/electron capture detection (GC/ECD). The capacity for aldehydes with one carbonyl group was 30-35 mumoles, and 15 mumoles for the dialdehyde, glutaraldehyde. The experimental sampling rates in mL/min were 8.86 +/- 0.38, acetaldehyde; 11.69 +/- 0.32, chloroacetaldehyde; 7.85 +/- 0.19, crotonaldehyde; 9.97 +/- 0.10, formaldehyde; 6.47 +/- 0.42, furfural; and 4.46 +/- 0.15, glutaraldehyde. Other data on valeraldehyde and acrolein have shown that the sampling constants were independent of face velocity between 0.1 to 0.35 m/s (20 to 70 fpm), temperatures between 9 to 48 degrees C, RH between 3 to 79 percent, and intermittent sampling exposure pattern. PMID- 10457650 TI - Certified reference materials for quality control of mercury and selenium determination in food. AB - Several certified reference materials were used for quality control of mercury and selenium determination in food through different analytical procedures and techniques. The materials were selected with respect to different composition of matrix and levels of certified values. The results agreed well with the certified values for all analysed materials. PMID- 10457651 TI - Determination of ethanol in chocolate shell pralines and filled chocolates by capillary gas chromatography. AB - The paper describes a simple gas chromatographic method for quantification of ethanol in distillates of chocolate shell pralines and fillings. The samples were prepared in two steps. The first step consisted of ethanol distillation from the product and the second involved capillary gas chromatography of 10% v/v distillate with expected ethanol content between 0.06% and 2.5% w/w. Quantification was carried out using iso-propanol as internal standard. The range of linear method response was 0.05-3.16% w/w of ethanol, which corresponded to products with ethanol content between 0.5 and 31.6% w/w. The detection limit was 0.0158% w/w and the quantification limit was 0.058% w/w of ethanol with the relative standard deviation of 2.5%. PMID- 10457652 TI - Wageningen Evaluating Programmes for Analytical Laboratories (WEPAL). AB - The paper describes three of the Wageningen Evaluating Programmes for Analytical Laboratories (WEPAL). These include the analyses of numerous compounds and elements and different parameters such as inorganic chemical composition, organic matter, polycyclic hydrocarbons (PAH), polychlorinated biphenyls (PCB), organochlorine pesticides, some herbicides, heavy metals, particle size, and so on in soil, sediment, compost, manure, and sludge. One programme includes the analysis of inorganic chemical composition, nutritional values, and selected vitamins and amino acids in plant samples. Finally, the paper describes how the results are reported and statistically evaluated. PMID- 10457653 TI - Regulations of the European Union for mycotoxins in foods. AB - Mycotoxins are fungal metabolites that are present in a large part of the world food supply and bear potential threat to food safety. The European Union has been working for several years now on the harmonization of some mycotoxin standards for foods based on toxicological evaluation by its Scientific Committee on Food (SCF). These standards will follow the ALARA (as low as reasonably achievable) principle and will be applied to aflatoxins in the first place followed by ochratoxin A, patulin, and several toxins from Fusarium spp. Detailed regulations (limits, methods of sampling, and analysis) for aflatoxins B, G, and M on the commodities at stake have been worked out. At the moment, a variety of interrelated approaches as to the intake and toxicity of ochratoxin A are being discussed to assess the hazards and to determine the associated risk of this toxin. The other toxins like patulin and the ones derived from Fusarium spp. such as zearalenon and fumonisins will be discussed in the years to come. PMID- 10457654 TI - Accreditation of analytical laboratories and proficiency testing: experiences from Thailand on the issues of pesticide analysis. AB - Thailand, the economic growth of which largely depends on agriculture-based industry, recognizes the importance of international standards for harmonization of international food trade and for protection of human health from chemicals used as food additives or those present as contaminants. Pesticides which are widely used in Thailand are the anticholinesterase pesticides, pyrethroids, coumarin derivatives, bipyridinium salts, and to some extent organochlorine compounds such as DDT in malaria control. Proficiency testing and accreditation of laboratories are recognized as effective means to ensure quality and validity of data and to enable adequate risk assessment of pesticide residues in food, work environment and environment in general. The paper is a synthesis of relevant local reviews and in-depth interviews with experts in the area of pesticide laboratory accreditation and proficiency testing. The paper refers to other schemes for quality assurance such as ISO guide 25, ISO 9000, and ISO 14000, and addresses future prospects of ongoing activities such as accreditation of inspection bodies in the area of industrial chemicals and occupational safety and health. PMID- 10457655 TI - Proficiency testing in the food microbiology laboratory. AB - Laboratories offering a quality service for the microbiological examination of food must implement a quality assurance system. An effective system should, in addition to daily quality control of procedures, consumables, and equipment, include external laboratory accreditation to a recognised standard, the use of validated test methods, and participation in an external quality assessment (proficiency testing) scheme. External quality assessment (EQA) is a system in which samples of known but undisclosed content are introduced into a laboratory's routine testing procedure, in other words, a challenge to those procedures. There are a number of EQA schemes available to food microbiology laboratories, either offering freeze dried mixtures of organisms as simulated foods or preweighed portions of spiked dried foods. There are a number of important requirements for an effective EQA scheme and a range of benefits from participation to a laboratory. These are discussed in the light of experience with the development of a specific scheme. PMID- 10457656 TI - [Megamin, faith, hope and placebos---a critical review]. AB - Megamin is a registered nutriceutical in Croatia. Allegedly, it consists mostly of a naturally occurring zeolite (aluminum silicate) ion-exchange material tagged with vitamins and minerals. Mass-media claimed that Megamin possessed strong anticancer properties although it had not been tested through an appropriate, randomized, double-blind, placebo-controlled, crossover clinical trial. The paper uses Megamin as a paradigmatic model to discuss how higher neurocognitive functions like hope and faith may be related to the placebo effect and give an illusion of symptomatic well being without supporting evidence in the objective signs of the disease. The only plausible role of Megamin is that it reduces gastrointestinal toxic burden by affecting the anaerobic fermenting processes after digestion of food and by removing harmful metabolites after medical treatment of cancer and/or liver and kidney organ failure. The psychological effect of unwarranted mass-media claims about the beneficial role of Megamin in cancer treatment is scrutinized. PMID- 10457657 TI - Nonneoplastic disorders of the brain. AB - Computed tomography (CT) and magnetic resonance imaging (MRI) are helpful in the diagnosis of many nonneoplastic brain disorders in the dog and cat. The ability of CT and MRI to depict normal and abnormal anatomy facilitates the identification of developmental anomalies, including hydrocephalus, Chiari malformations, arachnoid cysts, and cerebellar hypoplasia. These imaging modalities also allow the detection of hemorrhage and infarction and are therefore useful in the evaluation of spontaneous cerebrovascular disorders and head trauma. Finally, many inflammatory diseases, such as encephalitis, brain abscess, and parasite migration, cause abnormalities detectable by CT and MRI. Although more research on the imaging features of specific nonneoplastic brain disorders is needed, current information indicates that CT and MRI are useful in the management of these disorders. PMID- 10457658 TI - The spine. AB - Computed tomography (CT) and magnetic resonance imaging (MRI) are becoming increasingly accessible in veterinary medicine. Because of their ability to image the bony spine and spinal cord noninvasively and with exceptional detail, these techniques have revolutionized the way spinal disorders are diagnosed in both people and animals. Although the veterinary literature on this topic is limited, interpretation of CT and MR images of the spine are facilitated by the similarity of human and animal disease processes. This article provides an overview of imaging strategies, normal anatomy, and the CT and MRI features of degenerative, infectious, neoplastic, and vascular diseases of the spine. PMID- 10457659 TI - The eye and orbit. AB - Computed tomography (CT) and magnetic resonance imaging (MRI) provide excellent morphological detail of the eye and its associated structures. In veterinary medicine, these imaging modalities are most often used to diagnose and determine the extent of ocular or periocular tumors. They may also be used to diagnose inflammatory conditions of the orbital region and to determine the severity and extent of ocular trauma. This article reviews the applications of CT and MRI for ocular and orbital diseases, discusses normal findings, and presents representative case examples. PMID- 10457660 TI - The head: excluding the brain and orbit. AB - Applications for computed tomography (CT) and magnetic resonance imaging (MRI) scans of the head (excluding the brain and orbit) include the evaluation of neoplastic, inflammatory, and traumatic disorders of the skull (including the cranium, maxilla, mandible, and temporomandibular joints); of the nasal and oral cavities; of the nasopharynx and paranasal sinuses; and of the tympanic bullae. CT and MRI provide information not obtained with routine radiography and should be considered whenever advanced surgical or radiation therapy is anticipated. In this article, the role of CT and MRI in the diagnosis of head disorders will be reviewed and illustrated with case examples. PMID- 10457661 TI - The elbow. AB - Computed tomography (CT) and magnetic resonance imaging (MRI) are noninvasive methods of imaging the canine cubital joint. CT images are typically acquired using contiguous 1-mm slices, a bone reconstruction algorithm, and a field of view large enough to scan both elbows simultaneously. CT provides a detailed assessment of the medial coronoid process (MCP), radial incisure, anconeal process, and trochlear notch of the ulna; the humeral condyle; joint congruity; and osteoarthrosis. With MRI, use of a surface coil and 3D Fourier transformation gradient echo fast imaging sequences allow contiguous thin slices to be obtained. Both imaging techniques appear to be highly effective in the evaluation of elbow dysplasia, particularly for the detection of MCP fragmentation, although MRI is superior to CT for identifying nonmineralized cartilaginous fragments. PMID- 10457662 TI - VTEC O157 outbreak associated with a farm visitor centre in North Wales--update. PMID- 10457663 TI - Guidance on serological testing for syphilis. PMID- 10457664 TI - Over-the-counter yellow ultraviolet light protective lenses: any benefit for individuals with diabetes mellitus? AB - The use of yellow UV protective lenses did improve some individual's ability to see even under artificial and controlled circumstances. Each test, although given in random order, cannot be interpreted to demonstrate the full range of benefit. The improvement for most was modest. Any improvement was most likely due to increased contrast. Light of a short wavelength is scattered more than long wavelength light. The yellow UV protective lenses block light of a short wavelength, thus reducing scatter and increasing contrast. Therefore, patients with visual problems of increased scatter would be expected to demonstrate the greatest improvement. We did not test for the duration of benefit. Subjective reports, from other patients, who have routinely used these lenses (only yellow lens-naive patients were included in the trial) suggest that the benefit increased with duration of use. Patients who routinely use the yellow UV protective lenses state that due to the increased contrast, they squint less. This seems to be most true at dusk. These regular users note that both their eyes are less tired and driving, in particular, is less stressful with the use of yellow lenses. The small benefit might conceivably be magnified in a real world setting. Given that the lenses cost only $10 to $15 and can be purchased in any sporting goods store, even our small measured improvement is likely to be worthwhile. Finally, patients were tested with "off-the-shelf" yellow lenses. To benefit patients the study was designed for their convenience and the low purchase price. The color of the lenses can be chosen to maximize the desired effect. We did not test various wavelength yellow lenses. Consequently, there may be better, albeit more expensive, yellow/orange lenses which might be designed explicitly for this purpose. In summary, the use of these yellow "sunglasses" might provide some improvement in sight for diabetic patients while keeping expense to a minimum. PMID- 10457665 TI - Extremity pain in a child: a case report of leukemia. AB - Early clinical diagnosis of leukemia in a child is difficult. Leukemia must be considered in the differential diagnosis of extremity pain in children, especially if the "bone" pain is out of proportion to a suspected etiology. Initial laboratory studies, radiographs, and scintigraphy may be normal. Indistinct radiographic findings of periosteal reaction, osteolytic lesions, sclerotic lesions, osteopenia, or leukemic lines in a child older than two years should alert a physician to the possibility of leukemia. Clinical suspicion of leukemia should be piqued if bone scan results seem to be inconsistent with a suspected diagnosis or unanticipated sites of increased metabolic activity are found in the lower extremities. If leukemia is probable, a bone marrow aspiration may be needed to confirm the diagnosis. PMID- 10457666 TI - Continuing Medical Education (CME) on the Internet. PMID- 10457667 TI - Prostate cancer screening: what have we learned? PMID- 10457668 TI - The whole wide world is watching... PMID- 10457669 TI - Supporting our friends, the American Public Health Association, Vision Care Section. PMID- 10457670 TI - Juxtapapillary nerve fiber layer infarction as a complication of coronary artery bypass surgery. AB - BACKGROUND: Recent studies have detailed the prevalence and etiology of ocular complications resulting from coronary artery bypass surgery. Of these, retinal nerve fiber layer infarctions are reported most commonly. The clinical sequelae of nerve infarction may include loss of visual acuity, compromised pupillary function, and visual-field defects (the severity of which may be correlated with the location and extent of the insulted tissue). METHODS: A patient who had experienced bilateral juxtapapillary nerve fiber layer infarction with subsequent loss of visual acuity and peripheral visual field was followed postoperatively for more than 6 weeks. Immediately before our examination, he underwent quadruple coronary artery bypass graft surgery. RESULTS: From the data collected during initial and follow-up examinations, it was determined that the nerve fiber layer infarction was probably the result of a systemic ischemic event during an otherwise uncomplicated surgery. Such events may include hypovolemic blood loss, systemic hypotension during or following surgery, or a host of complications that would prevent adequate perfusion to capillaries in select regions of the eye. CONCLUSION: The pathology of bilateral juxatapapillary nerve fiber layer infarction as a result of substantial transient systemic ischemia may be explained by examining the microcirculation of this region of the retina. Due to certain anatomic and physiologic characteristics, the capillaries supplying the peripapillary zone are most susceptible to arterial vascular events such as ischemia. It is important to recognize this clinical presentation in order to rule out other possible causes for decreased visual acuity and field defects in the postoperative coronary bypass surgery patient. PMID- 10457671 TI - Weber's syndrome secondary to subarachnoid hemorrhage. AB - BACKGROUND: Since a large intracranial hemorrhage is a space-occupying mass, it may cause the brain to shift, resulting in neurologic deficits both at the location of the bleeding and at a site distal to the hemorrhage. A parietal lobe hemorrhage may push the brain downward, causing the uncus of the temporal lobe to herniate through the tentorial notch, compressing the midbrain. The signs of parietal lobe damage, uncal herniation, and several midbrain syndromes that effect ocular motility are discussed. CASE REPORT: A 66-year-old Hispanic man came to us with a history of a subarachnoid hemorrhage that involved the right parietal lobe. Several signs of damage to both the right parietal lobe and midbrain were evident, including an ipsilateral third nerve paresis with contralateral hemiplegia, Weber's syndrome. CONCLUSION: A patient who survives a subarachnoid hemorrhage may demonstrate permanent residual neurologic deficits subsequent to the acute event. The presentation is particularly complex when the hemorrhage is large and damage occurs at multiple locations. Damage at the level of the midbrain is evident when the findings include Weber's syndrome, which is one of several syndromes that involves the oculomotor nerve. PMID- 10457672 TI - Contact lenses after LASIK. AB - BACKGROUND: The corneal shape is significantly altered after LASIK, necessitating new contact lens designs and fitting techniques for optimal postoperative contact lens performance. METHODS: Procedures for fitting soft and rigid gas-permeable lenses are described on the basis of the authors' experience. Topographic map options are explored to assess the best curvature and contour data to assist in RGP lens fitting. RESULTS: Standard soft and rigid lenses are often used with success after LASIK. When fitting standard and reverse geometry RGP lenses, axial topographic maps provide the best curvature values to assist in the fitting of RGP lenses. CONCLUSIONS: Although patients may be apprehensive to pursue contact lens fitting after elective LASIK refractive surgery, new fitting techniques should turn a potentially difficult fitting into a teachable science. PMID- 10457673 TI - Juvenile onset primary open-angle glaucoma: three case studies and review. AB - BACKGROUND: Common clinical characteristics of juvenile onset primary open-angle glaucoma (JPOAG) include increased intraocular pressure, optic nervehead damage, visual-field loss, and a normal-appearing iridocorneal angle by gonioscopy. Histologic analysis of the angle structures may show varying developmental abnormalities that are observed to be less obvious as age of onset increases. Individuals who are African-American, male in gender, and myopio, are at highest risk--particularly if a positive family history exists. A genetic locus for juvenile onset primary open-angle glaucoma has been isolated to the long arm of chromosome 1. CASE REPORTS: Three separate cases of JPOAG diagnosed in a 16-year old African-American girl, a 9-year-old African-American boy, and a 28-year-old African-American woman are presented and reviewed. The symptoms, clinical presentation, disease progression, and treatment options are discussed in detail. CONCLUSION: Early recognition of this disease is possible through screening on all patients--particularly those who exhibit increased cupping with increased intraocular pressures and risk factors, such as youthful age, male gender, myopic refractive error, and African-American heritage with a positive family history of glaucoma. Family members should also be examined. PMID- 10457674 TI - You say you want a resolution... PMID- 10457675 TI - ARMD is robbing older people blind--and stealing their independence, too. PMID- 10457676 TI - Elevated disks. PMID- 10457677 TI - Aniseikonia. PMID- 10457678 TI - A protocol for the evaluation and treatment of atrophic age-related macular degeneration. AB - BACKGROUND: Age-related macular degeneration (ARMD), the leading cause of vision loss in aging Western societies, is (in part) a nutrition-responsive disease that can be stabilized through environmental intervention. The atrophic form of the disease constitutes nearly 90% of cases, yet a standardized ophthalmic and systemic evaluation protocol does not exist. METHODS: A basic science and epidemiologic rationale for evaluating atrophic ARMD using a standardized low cost, "low-technology," clinical approach is presented. The protocol is based on optometric physiological optics and ARMD retinal physiology/pathophysiology. RESULTS: The ophthalmic component involves baseline and serial measurements of contrast sensitivity (CSF), low-luminance, low-contrast visual acuity (SKILL), and glare recovery (GR) in each eye. The systemic component includes self administered evaluation of nutritional status (food and supplement intake), exercise and activities of daily living associated with ARMD (night driving/glare adaptation disturbance and dark green-leafy vegetable/plant food consumption). CONCLUSIONS: The literature supports careful baseline and serial evaluation of high-risk patients using germane parameters of ocular function and systemic health specific to ARMD. This "ARMD workup" is analogous to a glaucoma workup, but involves less equipment and chair time. Any optometrist can obtain these measurements using inexpensive testing protocols. (Part II of this paper presents serial environmental intervention case report data.) PMID- 10457679 TI - ARMD--pilot (case series) environmental intervention data. AB - BACKGROUND: A description and justification for using a standardized clinical low cost, "low-technology," ocular and systemic "ARMD work-up" protocol was presented in the methods paper (Part I). METHODS: Fourteen male patients (70 +/- 9 years), receiving 0.73 +/- 0.45 portions of dark-green, leafy vegetables/day base intake, were placed on an additional portion of 5 ounces sauted spinach 4 to 7 times per week or lutein-based antioxidant (three patients). Detailed "ARMD work-up" data for six of these patients and summary pilot (case series) environmental intervention data are presented. RESULTS: One, Patients demonstrated short-term positive effects in visual function in one or both eyes with this mild therapeutic approach: Amsler grid (87%); Snellen Acuity (71%); Contrast sensitivity (92%); SKILL (65%); Glare recovery (69%); and Activities of Daily Vision Subscale (60%); Two, There was no obvious correlation between ophthalmoscopic appearance of the retina and visual outcome; and Three, Patient symptoms did not necessarily correlate with observed changes in visual function. CONCLUSIONS: The approach to atrophic ARMD presented here warrants informal practitioner replication and formal randomized prospective clinical case-control evaluation. PMID- 10457680 TI - Dietary prevention of age-related macular degeneration. AB - BACKGROUND: Age-related macular degeneration (ARMD) is the leading cause of blindness in people over the age of 65, and the prevalence of ARMD is expected to increase as the population ages. Although the incidence of ARMD increases sharply with age, recent studies indicate that prevention measures and dietary changes, implemented early in life, can reduce an individual's risk of ARMD. METHODS: Several dietary components have been proposed and studied with regard to their ability to protect against ARMD; these components include antioxidant vitamins and specific carotenoids. In particular, consumption of dark green, leafy vegetables has been shown in clinical studies to reduce the risk of ARMD. RESULTS: Biochemical studies of such vegetables have found that they contain several nutrients that may account for this effect, including high concentrations of the related carotenoids lutein and zeaxanthin. Structural and clinical studies have shown that these carotenoids are concentrated in the retinal macular pigment and that such accumulation is dependent on dietary intake. Further studies have indicated that the density of the macular pigment is related to preservation of visual sensitivity and (possibly) protection from ARMD. CONCLUSIONS: Large-scale clinical trials will be necessary to demonstrate that specific agents can reduce the incidence of ARMD. Nevertheless, specific dietary components--particularly, the carotenoids found in dark green, leafy vegetables--have shown great promise. While lifestyle modifications such as smoking cessation, reduction of alcohol consumption, and the wearing of sunglasses may reduce the risk of ARMD, it is likely that consumption of specific dietary components can reduce the risk further. PMID- 10457681 TI - Providing timely and ongoing vision rehabilitation services for the diabetic patient with irreversible vision loss from diabetic retinopathy. AB - BACKGROUND: Diabetic retinopathy (DR) remains the leading contributor to severe vision loss in the United States among persons 20 to 70 years of age. Despite advances in disease management and treatment, patients with vision loss from DR continue to constitute a significant portion of patients served in vision rehabilitation service (VRS) settings. These patients present special challenges to VRS providers because of early onset, fluctuations in and the complex nature of vision loss, unique visual demands of disease management, and associated multi system losses. CASE REPORTS: After introductory epidemiologic review, a case presentation format is used to illustrate solutions a multidisciplinary VRS can offer the special visual challenges of the person with diabetes with vision loss from DR. Four patients are presented--ages 30 to 70 years--with varying degrees and types of vision loss, with different lifestyle demands and disease management needs. The cases address vocational issues, vision fluctuation, coordinating adaptive solutions to complex visual losses, and meeting diabetic needs to measure medication, insulin, and blood glucose levels, to maintain skin care, diet, exercise, transportation, family roles, and support systems. CONCLUSIONS: The unique and complex needs of people with diabetes who experience vision loss can be well addressed through timely and ongoing VRS consultations, in conjunction with medical/ocular disease management. PMID- 10457682 TI - The AOA Sports Vision Section: a valuable membership benefit. PMID- 10457683 TI - Dysthymia: a common mild depression. PMID- 10457684 TI - Eye on the ball: an interview with Dr. C. Stephen Johnson and Mark McGwire. Interview by J. David Kopp. PMID- 10457685 TI - Injury prevention: where do we go from here? AB - BACKGROUND: Most eye injuries are preventable. Military personnel, workers, athletes, and other spectacle wearers--especially children and the functionally one-eyed--who require protection from impact, should expect that safety eye-wear actually protects. PURPOSE: To present to eye care professionals the current state-of-the-art in eye injury prevention. METHODS: A review of the current eye protection standards, guidelines, and warnings for the activities of daily living, work, hobbies, education, and sports with emphasis on the importance of standards and the role of the recently organized Protective Eyewear Certification Council (PECC). CONCLUSIONS: The prescriber and dispenser are obliged to prescribe, fabricate, and dispense safe and effective eyewear. PECC will help the eye care professional fulfill this obligation. PMID- 10457686 TI - Optometry and ophthalmology relations in managed care. AB - BACKGROUND: Optometrists and Ophthalmologists have a history of difficult professional relationships. This paper discusses common issues both professions face while working under managed care. CONCLUSIONS: Managed care initiatives may help to improve the ability of Optomotrists and Ophthalmologists to work together to treat the patient's eye problems. PMID- 10457687 TI - Comparison between anti-reflection-coated and uncoated spectacle lenses for presbyopic highway patrol troopers. AB - BACKGROUND: The use of anti-reflection coatings (AR) for spectacle lenses is steadily increasing. These ultra-thin coatings reduce reflections in lenses, increasing light transmittance about 8% (from 91% to 99%). Patient reports of crisper detail and brighter environment indicate that this perceived increase in visual function would probably be of benefit to many, including highway patrolmen who must make daily judgments on the basis of fine visual detail. METHODS: Nineteen presbyopic Missouri State Highway Patrolmen (MSHP) participated in this crossover study. Subjects randomly wore Varilux Comfort progressive addition lenses in CR-39 that were uncoated or AR-coated. The AR coating was Crizal. Subjects wore each lens for 1 month, then compared the lenses directly for 1 week. Data were collected for lens scoring (both general and job specific) and lens preferences. RESULTS: The AR-coated lens was found to be significantly better in most categories. Reduction in glare and reflections during the day, night, indoors, and in the patrol vehicle--as well as improvement in overall night vision--were highly significant. Eighty-nine percent of the subjects chose the Crizal-coated lenses as their overall preferred lens. CONCLUSIONS: The results of this study appear to demonstrate a noticeable subjective preference for AR-coated lenses when used by a group engaged in a visually demanding occupation. PMID- 10457688 TI - Accommodative facility test results and academic success in Polish second graders. AB - BACKGROUND: Accommodative infacility, as commonly measured by accommodative flippers, has been implicated as a factor in academic underperformance. This study compares four areas of academics (reading, writing, math and gym) to accommodative flexibility scores. METHODS: Seventy-six elementary school children with a mean age was 8 years, 8 months were tested monocularly and binocularly with traditional accommodative flexibility flipper testing and with a new accommodative flexibility apparatus that allows control of visual acuteness, minification/magnification, and reaction time. These scores were then compared with academic scores using a number of failure criteria. The academic ratings were based on teacher responses for each student. RESULTS: Our data did not show any clear correlation or relationship between evaluations by reading, writing, math, or gym teachers and accommodative flexibility by either the traditional or new testing methods. CONCLUSION: Accommodative function, free of contaminating variables, does not appear to predict academic function any better than the traditional. PMID- 10457689 TI - Monovision and LASIK. AB - BACKGROUND: Many LASIK patients are early and mid-presbyopes. Monovision is commonly targeted for these patients because of demographics. Considerations are reduced stereopsis, reduced acuity, night driving, and liability. CASE REPORTS: Case studies are used to illustrate contact lens and alternative trials for presbyopic and prepresbopic monovision LASIK patients. CONCLUSION: A preoperative monovision contact lens trial can be used to demonstrate monovision effects to the candidate for refractive surgery. Selection of add power, selection of distance and near eye, residual astigmatism, management, and gradual transistioning with regards to adaptation are discussed. PMID- 10457690 TI - Familial Duane's retraction syndrome. AB - BACKGROUND: Duane's retraction syndrome (DRS) has been described as a clinical entity for more than a century. Although the majority of cases occur spontaneously, some cases of DRS are inherited. CASE REPORTS: A young Romanian boy with a known left abduction deficit was determined to have DRS. On thorough examination of other family members, it was discovered that his younger brother and father also had the syndrome. Clinical findings of the affected family members are described. RESULTS: All three family members showed left esotropia in primary gaze, along with a compromised ability to turn the left eye outward. They also demonstrated an anomalous head turn to the left, and mild narrowing of the palpebral fissure and retraction of the globe on adduction. These are all classic signs of unilateral DRS. CONCLUSION: These findings provide further evidence of an autosomal dominant mode of inheritance in some cases of DRS. PMID- 10457691 TI - A profession's commitment to increased public service: optometry's remarkable story. PMID- 10457693 TI - The influence of gender on the ocular surface. AB - PURPOSE: Dry eye is more prevalent among women than men. To examine whether gender influences the ocular surface, conjunctival goblet-cell density was monitored for a 30-day period. Changes in goblet-cell density are a sensitive marker for ocular surface disease. METHODS: Six men and twelve women who were free from ocular surface disease were sampled by impression cytology on the inferior bulbar conjunctiva every other day for 30 days. Samples were analyzed for goblet-cell content. The mean age of the subjects was 24 years for the men and 25.4 years for the women. A minimum of 200 epithelial cells were scored per microscopic field examined. RESULTS: There is a statistically significant difference in goblet-cell count between men and women over the 30-day test period. The mean goblet-cell count measured for men was 3.75% +/- 1.04%, while the mean count for women was 2.545% +/- 0.8%. When the data on women were replotted with reference to the menstrual cycle, the largest and longest depression of goblet-cell count occurs midcycle, around the time of ovulation. A significant difference in goblet-cell count was also noted when subjects using oral contraceptives (3.065% +/- 0.98%) are compared with those not using oral contraceptives (2.28% +/- 0.92%). CONCLUSION: It appears there may be a reproductive hormonal influence on conjunctival goblet-cell count. PMID- 10457692 TI - Comparisons of eye movements before and after a speed-reading course. AB - BACKGROUND: The techniques of speed reading are widely used and generally accepted, but few studies have investigated whether- and how-speed reading actually improves reading ability in the typical reader. METHODS: Using the Ober2, an infrared monitoring device that accurately tracks eye movements, we measured the eye movements of 59 students. The Ober2 calculates reading speed, number of fixations per 100 words, number of words seen in each fixation, number of regressions per 100 words, and duration of fixation. Comprehension was assessed with ten detailed true or false questions. RESULTS: One group of the students (n = 25) participated in a speed reading class; the other group (n = 34) did not. After completion of the course, all 59 were re-measured on the Ober2. The speed-reading group improved significantly in five of the six aspects tested. Comprehension for the speed reading group showed an insignificant decrease. CONCLUSIONS: We conclude that measurable changes in reading eye movements accompany successful completion of a speed-reading course. PMID- 10457694 TI - Relationship of impression cytology and tear ferning to reports of dry eye. AB - PURPOSE: This study investigated whether there is a significant correlation between mild-to-moderate patient reports of dry eye and results on the tear ferning and impression cytology tests. METHODS: One hundred and four subjects were surveyed for presence or absence of dry eye symptoms. Membrane impression cytology and tear ferning were performed on each subject. Impression cytology specimens were evaluated for the presence of goblet cells, presence of epithelial cell sheets, and epithelial cell morphology. Tear specimens were evaluated for quality of ferning. RESULTS: A significant correlation was found between subjective dry eye symptoms and impression cytology results (cc = 0.28, p = 0.003). However, there was no statistically significant correlation between dry eye symptoms and tear fern results (cc = 0.072, p = 0.458). CONCLUSIONS: Impression cytology appeared to be a better predictor of mild-to-moderate dry eye symptoms than tear ferning. PMID- 10457695 TI - Crack eye syndrome. AB - BACKGROUND: Cocaine is an alkaloid prepared from the leaves of the Erythroxylon coca plant. It is widely recognized as one of the most dangerous illicit drugs in use today. The U.S. Substance Abuse and Mental Health Services Administration estimates that 23.5 million Americans have used cocaine at some time in their life. Corneal defects from crack cocaine were first described in 1989 and later named crack eye syndrome in 1993. CASE REPORT: A 40-year-old man reported to the eye clinic reporting vigorous eye rubbing after repeated exposure to and use of crack cocaine. A corneal infiltrate with an overlying epithelial defect developed and the man was treated with ciprofloxacin, homatropine, and diclofenac. This case presents background information concerning the systemic and ocular manifestations of cocaine, as well as the clinical presentations of crack eye syndrome, with recommendations on treatment. CONCLUSIONS: A thorough social history should be elicited when patients who have unilateral or bilateral corneal defects of unknown origin are examined, or when treating persons with no other known risk factors for corneal disruption. Prompt recognition and treatment are the main factors in successful management of crack eye syndrome. PMID- 10457696 TI - From a hyperope's point of view. PMID- 10457697 TI - A clinical review of hyperopia in young children. The Hyperopic Infants' Study Group, THIS Group. AB - BACKGROUND: Hyperopia is the most common refractive error of children. Children with mild (or even moderate) levels of hyperopia usually do not experience visual problems resulting from this hyperopia. However, children with moderate-to-high degrees of hyperopia are at significantly increased risk for the development of amblyopia and strabismus. It is this association with these visually threatening disorders that makes hyperopia in children an important public health problem. In addition, even lesser degrees of hyperopia may affect the child's ability to perform well in near-related tasks, such as reading. The effect hyperopia has on an individual child is dependent on a variety of factors, including the magnitude of hyperopia, the age of the individual, the status of the accommodative and convergence system, and the demands placed on the visual system. Early detection and treatment of hyperopia may help prevention of potential complications from adversely impacting the child's vision. Although much is known about childhood hyperopia and its effects on vision, there is also much that is not known. The natural history, ocular biometry, relationship to accommodative function, the indications for treatment, and the most effective treatment modalities are among the underlying issues and clinical considerations awaiting more complete understanding. PMID- 10457698 TI - Efficacy of vision therapy for convergence insufficiency in an adult male population. AB - BACKGROUND: Although vision therapy has reportedly been very successful in elimination of asthenopic symptoms in adults with convergence insufficiency, controlled studies have not been performed, and a clinical bias exists against prescribing vision therapy for adults with convergence insufficiency. METHODS: Sixty adult males over the age of 40 years (median age, 65 years) with convergence insufficiency were divided into three treatment groups: office-based vision therapy with supplementary home therapy, home therapy only, and a control group. RESULTS: Vision therapy was successful in 61.9% of patients who received in-office plus home therapy, in 30% of patients who received home therapy only, and in 10.5% of the control group. The success rate for patients who received active in-office vision therapy supplemented with home procedures was significantly greater than that for controls. Home therapy alone was less successful than in-office therapy. The success rate obtained with home therapy alone was not significantly greater than that demonstrated by controls. CONCLUSIONS: Vision therapy is effective in eliminating asthenopia and improving convergence function in adult patients. In-office therapy combined with home therapy tends to produce better results than does home therapy alone. PMID- 10457699 TI - Reversal of intraocular pressure increases with 0.5% apraclonidine after dilated fundus examination in patients with chronic open-angle glaucoma. AB - BACKGROUND: Apraclonidine 1.0% has been shown to reverse the potential intraocular pressure (IOP) increase after pupil dilation IOP increases in patients with chronic open-angle glaucoma. However, it is only approved for preventing IOP spikes after laser surgery. The purpose of this study is to determine the effectiveness of 0.5% apraclonidine in reversing IOP increases after pupillary dilation in patients with chronic open-angle glaucoma. METHODS: Twenty-two patients with chronic open-angle glaucoma were found to have an increase in post-dilation IOP of at least 4 mmHg from pre-dilated levels (baseline) in both eyes. IOP was measured 1 hour after dilation, after which two drops of 0.5% apraclonidine were instilled in one eye and the IOP was remeasured 15 minutes later in both eyes. Instillation of 0.5% apraclonidine in one eye was continued every 15 minutes and IOP was measured 15 minutes after each instillation, until the pressure returned to baseline levels. RESULTS: The IOP of the initially treated eye of all 22 patients returned to within levels clinically insignificant from baseline IOP within 90 minutes. By comparison, the IOP of the control group (untreated eye) remained elevated. Once the initial treatment eye returned to baseline levels, the control group was then treated with 0.5% apraclonidine, resulting in a lowering effect of the IOP in similar fashion to the initial treated group. CONCLUSIONS: Apraclonidine 0.5% appears to be effective in reduction of post-dilated IOP increases in patients with chronic open-angle glaucoma. PMID- 10457700 TI - A comparison of the Synemed Glaucoma and the Humphrey 30-2 threshold perimetry tests. AB - BACKGROUND: Automated perimeters (in general) are similar; however, caution may be exercised when visual field results from two different instruments are compared. The purpose of this study was to compare threshold measurements in the central field between the Synemed (Optifield 1) Glaucoma Test and the Humphrey 30 2 test in a young patient population. METHODS: One hundred twenty subjects were tested. The subjects were selected according to specific criteria considering ocular and systemic disease and refractive error. The ages ranged from 15 to 35 years, with a mean age of 25.9 years. The central 30 glaucoma test was used with the Synemed instrument, and the 30-2 test was used with the Humphrey instrument. Mean threshold values for the entire field, hemifields, quadrants, and sectors were compared between instruments. RESULTS: The difference in the mean value for the collective threshold values for each pattern for each instrument were calculated and compared. The difference between the instruments for the mean dB threshold value for each pattern was less than the expected short-term fluctuation and therefore are equivalent for clinical purposes. CONCLUSION: For the patterns examined here, the Synemed Optifield 1 and Humphrey Field Analyzer provided comparable results in this population. PMID- 10457701 TI - Ocular toxocariasis: a rare presentation of a posterior pole granuloma with an associated choroidal neovascular membrane. AB - BACKGROUND: Ocular toxocariasis is a rare infection caused by the nematode larvae of toxocara canis, which is commonly found in dogs. Human transmission is usually via geophagia, the ingestion of food contaminated with the toxocara eggs, or contact with infected puppies, often resulting in devastating ocular and/or systemic effects. Distribution is worldwide; however, a higher incidence is demonstrated in the United States. METHODS: A 17-year-old black woman sought treatment at a neighborhood health center with a report of gradual decrease in vision from her left eye over a 3-month period. Her ocular and systemic histories were unremarkable. Anterior segment evaluation revealed no signs of anterior uveitis. The posterior pole showed a 1.5 DD, round, raised, white, subretinal lesion adjacent to the fovea with an overlying serous retinal detachment and retinal hemorrhage. RESULTS: She was referred to a retinologist who performed both fluorescein and indocyanine green (ICG) angiographies. A serum toxocara ELISA test was also ordered. Fluorescein angiography revealed hyperfluorescence consistent with the granuloma. The ICG demonstrated an occult choroidal neovascular membrane (CNV) underlying the area of hemorrhage inferotemporal to the granuloma. CONCLUSION: This paper illustrates the case presentation and includes an extensive review of the ocular and systemic manifestations of toxocariases. A description of ICG videoangiography, therapeutic approaches, and management will also be discussed. PMID- 10457703 TI - Unmet need for eyeglasses: results from the 1994 Robert Wood Johnson Access to Care Survey. AB - BACKGROUND: Although poor access to general medical care services has been documented widely, the unmet need for supplemental health care services--such as eyeglasses--has been largely ignored. This article documents the inability to obtain eyeglasses on the national level and for various subpopulations using the 1994 Robert Wood Johnson Foundation National Access to Care Survey. METHODS: The Survey, a followup to the 1993 National Health Interview Survey, collected data on respondents' inability to obtain health care services due to access barriers. The sample comprised 3,480 observations, weighted to represent the U.S. civilian non-institutionalized population. RESULTS: More than 5% of the U.S. population reported an unmet need for eyeglasses, and of these, more than 80% cited financial reasons as the primary barrier. Persons in poor health and blacks were most likely to face barriers to obtaining eyeglasses. CONCLUSION: Understanding this under-served population and their unmet needs can help policymakers formulate new initiatives. The focus on preventive care within managed care organizations may ease some barriers for the insured population. Policy should focus on uninsured and underinsured working individuals who cannot afford and do not receive public assistance for needed eye care. PMID- 10457702 TI - Orbital fibrous histiocytoma: case report and literature review. AB - BACKGROUND: Fibrous histiocytomas are a diverse group of soft tissue tumors classified histiologically as benign, locally aggressive, and malignant. These tumors are found throughout the body, but seem to have an affinity for the periorbital area. They account for one percent of all ocular masses and are the most common primary mesenchymal tumor of the orbit. Associated ocular signs and symptoms include decreased visual acuity, proptosis, diplopia, pain, restricted extraocular muscle movement, swelling of the eyelids, and conjunctiva, as well as disk edema. CASE REPORT: A case of a benign orbital fibrous histiocytoma is presented. The patient reported intermittent pain and occasional diplopia; severe edema of the right upper eyelid; and proptosis and inferior vertical displacement of the right globe were observed. Computed tomography revealed a well-defined mass that was subsequently surgically removed and histopathological results from the Armed-Forces Pathology Institute confirmed the diagnosis. CONCLUSIONS: Orbital fibrous histiocytomas are rare periocular tumors that can manifest multiple ocular signs and symptoms. Careful histologic examination is necessary for diagnosis, since these tumors have a wide range of morphology. Differential diagnoses include orbital masses with similar radiologic or histologic findings. PMID- 10457704 TI - Laser pointers: a new problem facing law enforcement officers. PMID- 10457705 TI - Laser pointers: toys, nuisances, or significant eye hazards? AB - BACKGROUND: Laser pointers have been used inappropriately to harass and "dazzle" victims. Reports of retinal damage caused by pointers have also been circulated in the popular press. METHODS: Information on pointer abuse was collected from the literature and through discussions with specialists. RESULTS: Few, if any, documented cases of permanent retinal damage caused by laser pointers could be found. For actual damage to occur, viewing, times need to exceed approximately 10 seconds. Exposures of this duration would require the person being lased to cooperate by holding fixation on the laser beam. CONCLUSIONS: Although the risk of permanent retinal damage from a laser-pointer beam is minimal, other risks include dazzle, annoyance, and concern that a weapon-aiming device rather than a pointer is generating the laser beam. PMID- 10457706 TI - Postprandial transient visual loss. AB - BACKGROUND: Temporary loss of vision after ingestion of a large meal is a phenomenon that has been reported in only a small number of cases in the ophthalmic literature. CASE REPORT: A 60-year-old man reported episodes of blurred vision associated with visual aberrations in his left eye for 5 minutes after eating large meals. His medical history was significant for carotid artery disease, hypertension, "diet-controlled" diabetes, and hypercholesterolemia. The patient's symptoms abated when he ate in smaller quantities and was placed on a regimen of IOP-lowering eye drops. CONCLUSIONS: Postprandial transient visual loss can occur when blood gets shunted to the mesenteric system after eating, leaving the eye hypoperfused. It most commonly takes place in individuals whose ocular perfusion is already compromised by carotid disease, in combination with other vascular conditions that result in the inability to compensate for decreased perfusion to the eye. The signs and symptoms of postprandial visual loss--as well as the mechanisms thought to contribute to it--are discussed. PMID- 10457707 TI - The occurrence of ocular and visual dysfunctions in an acquired brain-injured patient sample. AB - BACKGROUND: Acquired brain injury (ABI) is a general term referring to brain injury occurring secondary to trauma, stroke, or post-surgical complications. This paper reports on the occurrence of various ocular and visual conditions in an ABI patient sample. METHODS: The subjects for this study were 62 brain-injured adults who resided in two extended care facilities in the New York area. Subjects ranged from 19 to 70 years of age. The standard protocol for the visual evaluation incorporated a patient interview; cover test; refraction; and assessment of visual acuity, ocular motility, accommodation, binocularity, visual fields, color vision, contrast sensitivity, pupils, and anterior and posterior segments. RESULTS: Results were reported as a ratio comparing the occurrence of a condition in our ABI sample relative to that in a reference normal population (where normative data are available) and as a comparison to other published data on ABI samples. An increased occurrence of exo deviations, oculomotor dysfunctions, and vertical deviations was evident. An elevated occurrence of dry eye, blepharitis, optic nerve pathologies, and visual field deficits was also manifest. CONCLUSION: Our patient sample demonstrates that certain ocular and visual conditions occur more frequently among ABI patients in comparison to a random, adult population. PMID- 10457708 TI - Indoor-air quality and ocular discomfort. AB - BACKGROUND: Almost 25 million workers in 1.2 million commercial buildings in the United States have symptoms of sick building syndrome (SBS). It is a source of ocular discomfort and may exacerbate the successful wear of contact lenses. This study examines the relationship between SBS and oculovisual discomfort. METHODS: Measurements of carbon dioxide, formaldehyde, temperature, and humidity were performed in 12 public office buildings that contained a total of 877 occupants. A questionnaire survey of the occupants was also performed. RESULTS: Thirty-five percent of the occupants of the buildings were dissatisfied with the indoor-air quality. Twenty-nine percent of the office building occupants reported ocular discomfort. CONCLUSION: There was a significant correlation between eye irritation, throat irritation, and symptoms of blurred near vision. Ocular discomfort could be an indicator of poor indoor-air quality and a sick building syndrome. PMID- 10457709 TI - The National Eye Institute outlines eye research goals. PMID- 10457710 TI - Case records of the Department of Medicine University of Mississippi Medical Center. 1) Malignant lymphoma, low-grade, polymorphic. T cell small lymphocytic lymphoma versus lymphomatoid granulomatosis, involving intestines, mesentery, omentum, refroperitoneal lymph nodes, lungs heart, liver, and spleen. 2) Malignant ascites. PMID- 10457711 TI - MisHIN: an outreach program to help practicing physicians keep up with medical literature. PMID- 10457712 TI - An interview with Todd Coulter, M.D., member-at-large on the governing council of the AMA Young Physicians Section. Interview by Karen A. Evers. PMID- 10457713 TI - The pitfalls of herbal medicines. PMID- 10457715 TI - The enhancement syndrome. PMID- 10457714 TI - Does syncope occur more often on sunday? PMID- 10457717 TI - Grand Rounds. Minnesota physicians on vacation. PMID- 10457716 TI - Bombarded by stress. Healthy habits to avert burnout. PMID- 10457718 TI - Time out. Physician sabbaticals can provide relief from burnout, a chance to learn new skills, and time for introspection. PMID- 10457719 TI - One pill makes you larger. The ethics of enhancement. PMID- 10457720 TI - Researchers find strain of E. coli that may cause miscarriages. PMID- 10457722 TI - Innocent victims. The connection between animal abuse and violence toward humans. PMID- 10457721 TI - Occupational hazards raise prostate cancer risk for farmers. PMID- 10457723 TI - Malpractice claims and the quest for perfection. PMID- 10457724 TI - About face. PMID- 10457725 TI - HIV homes tests. PMID- 10457726 TI - Clinical pharmaceutical research. PMID- 10457727 TI - Sickle cell patients find a brand New World. AB - As recently as the 1970s, sickle cell anemia patients had little hope of living past their teen years and little relief from painful crises. With the 1980s came the use of prophylactic penicillin to significantly reduce morbidity and mortality from infection in sickle cell youngsters. One of the sickle cell breakthroughs of the 1990s is the use of hydroxyurea for sickle cell treatment in adult patients. In the future, gene therapy may be the answer to the complex problems sickle cell anemia presents. PMID- 10457728 TI - H. pylori and peptic ulcers still misunderstood. AB - Heliocobactor pylori is the most common chronic infection in the world, affecting one-third of all Americans. It causes ulcer disease and is considered to be a class I carcinogen. It can be cured with antibiotics. The majority of ulcer patients are neither treated nor tested for this dangerous pathogen. What can be done to help these patients? PMID- 10457729 TI - What is New Jersey doing with the tobacco settlement money? PMID- 10457730 TI - Bloodless surgery. AB - Doctors and nurses in the forefront of the bloodless medicine and surgery movement say that virtually all hospitals now employ at least some facet of bloodless medicine. Because the strategies generally promote good health and save money, some New Jersey hospitals have even made bloodless medicine a standard of care regardless of patients' religious or moral convictions. PMID- 10457732 TI - Medical fraud. An ounce of compliance. PMID- 10457731 TI - Confronting Y2K physician practice guidelines. AB - The year 2000 computer problem is primarily a result of a programming technique called two-digit date processing. This technique reduced dates from eight digits to six digits, as programmers assumed the first two digits in every year would be 19. Software, hardware, and other computer equipment may not recognize or process the date properly when the year changes this coming January. PMID- 10457733 TI - Use of the 12-item short-form (SF-12) Health Survey in an Australian heart and stroke population. AB - The objective of this study was to validate the SF-12 Health Survey in heart and stroke patients using a community-based study. Between November 1995 and August 1996, 3,362 patients were invited to join the Hunter Heart and Stroke Register in New South Wales, Australia and to complete the SF-12 Health Survey. Of the 3,362 patients, 2,341 (70%) returned the SF-12. Of those 2,341 patients, 78% completed all 12 items. Those who did not complete the questionnaire were significantly more likely to be females, older, less educated, have stayed longer in hospital and been admitted on emergency. The SF-12 demonstrated construct validity in an analysis restricted to the 1,831 patients who completed the questionnaire: scores measuring physical and mental health status were statistically significantly higher in men than women, in younger than older, in those who had shorter than longer lengths of stay in hospital, in patients whose hospital admissions were planned than emergencies and in heart than stroke patients. Construct validity of the SF-12 among patients able to complete the SF-12 suggests considerable potential for its use in assessing health status in large-scale surveys. However, caution should be taken with the heart and stroke population because of a relatively high in completion rate. PMID- 10457734 TI - Comparison of Euroqol EQ-5D and SF-36 in patients with chronic fatigue syndrome. AB - The objective of the study was to compare the Euroqol EQ-5D (Euroqol) and short form 36 (SF-36) health questionnaires in patients with chronic fatigue syndrome (CFS). One hundred and twenty-seven outpatients referred to a hospital-based infectious disease clinic with a diagnosis of CFS were contacted by post and asked to complete both questionnaires. Additional data were determined from hospital casenotes. Eighty-five patients returned correctly completed questionnaires. Euroqol health values and visual analogue scale (VAS) scores were strongly and significantly correlated with all dimensions of the SF-36, with the exception of physical limitation of role. SF-36 dimensions were in turn strongly and significantly correlated with each other, with the same exception. Patients reported a high degree of physical disability and a moderate degree of emotional or psychological ill-health. The Euroqol elements dealing with mobility and self care referred to inappropriately severe degrees of disability for these patients with CFS. Similarly some dimensions in the SF-36 were oversensitive and did not discriminate between patients with moderate or severe disability. It was concluded that Euroqol scores correlated strongly with SF-36 scores and provided useful information about patients with CFS and that Euroqol would be a useful tool for the rapid assessment of health status in CFS. The current Euroqol instrument refers to inappropriately severe degrees of disability for patients with CFS and would need to be modified to be maximally useful in this situation. PMID- 10457735 TI - Towards a composite scoring solution for the Neurobehavioral Functioning Inventory. AB - Little research has been conducted towards the development and evaluation of a measure of quality of life specific to head/brain injury populations. Accordingly, we examined responses to the Neurobehavioral Functioning Inventory in the context of a clinical trial for head injury patients (n = 655) conducted in 14 countries. To reduce the 66 item scale into a smaller number of composite scales, principal components analysis was conducted. Scales were constructed assessing four categories of symptoms: cognitive deficits, depression, aggression and somatization. The internal reliabilities (alpha coefficient) of the four scales were generally acceptable (range = 0.79-0.92). Scores on all four scales correlated significantly with patient-rated overall quality of life and all but the aggression scale correlated significantly with overall clinical severity. The need for more formal evaluation of this and other disease-specific measures is discussed. PMID- 10457736 TI - Assessing the consistency of psychometric properties of the HRQoL scales within the EORTC QLQ-C30 across populations by means of the Mokken Scaling Model. AB - This paper demonstrates how the Mokken Scaling Model and other statistical tools may be useful in assessing the consistency of psychometric properties of health related quality of life (HRQoL) scales across various populations. The main focus is the psychometric performance of the scales proposed for the EORTC QLQ-C30 in seven patient groups totalling more than 2,000 cancer patients. All scales performed satisfactorily in the total sample with the exception of the role functioning and cognitive functioning scales, which failed in terms of reliability and item discriminant validity. The descriptive statistics for the scales show that several of them, particularly those that build upon only two items, have discrimination problems at the extremes, visible in the high percentages at the maximum or the minimum observed values. The scalability analysis in the subsamples showed that the essential assumption in the Mokken Scaling Model of equal item step order does not hold for the cognitive functioning, emotional functioning and physical functioning scales. We conclude that the Mokken Scaling Model is well suited to the purpose of examining the generalizability of HRQoL scales across subpopulations although a global statistical test of the fit of the measurement model is not available. PMID- 10457737 TI - Reliability, validity and responsiveness of two multiattribute utility measures in patients with chronic obstructive pulmonary disease. AB - The objective of the study was to evaluate the reliability, validity and responsiveness of two generic multiattribute methods for measuring utility for health states: a 15-dimensional (15D) and a five-dimensional method (EuroQol and EQ-TTO). A self-administered questionnaire with both measures was used in 59 outpatients with chronic obstructive pulmonary disease and the findings compared with standard gamble (SG) and time trade-off (TTO) utilities, spirometry and arterial blood gases. Quality of life scores were smallest for EQ-TTO (median 0.73) and highest for SG and TTO (median 0.91 and 0.95 respectively), while 15D gave intermediate values (median 0.80). The test-retest reliability over 14 days was: 15D (p = 0.90) and EQ-TTO (p = 0.73), using Spearman's rank correlation. 15D was better than EQ-TTO at discriminating between groups of patients after reported global rating of change over 12 months (P = 0.004 versus P = 0.09), indicating that 15D was more responsive. The 15D instrument has many attractive properties when compared to the EQ-TTO method, including a better reliability and responsiveness. Validity depends on validation method. The findings in this study indicate that the different utility measures measure different aspects of health related quality of life (HRQoL). Caution should be taken when choosing utility instruments in cost-utility studies, as this can strongly influence the results. PMID- 10457738 TI - The Caregiver Quality of Life Index-Cancer (CQOLC) scale: development and validation of an instrument to measure quality of life of the family caregiver of patients with cancer. AB - Family caregivers have become increasingly responsible for providing home care for cancer patients. Research has documented the distress of family caregivers; however, little has been done to evaluate the broader impact of caregiving on quality of life (QoL). The objectives of this study were to evaluate the reliability and validity of the Caregiver Quality of Life Index-Cancer (CQOLC), a new self-report instrument. Two hundred and sixty-three family caregivers of patients with lung, breast or prostate neoplasms participated. Test-retest reliability was 0.95 and internal consistency was 0.91. As expected, there were moderate correlations with overall mental health (r = 0.64), emotional distress (r = -0.50 to -0.52), burden (r = -0.65) and patient's performance status (r = 0.47) and low correlations with overall physical health (r = 0.13), social support (r = 0.22) and social desirability (r = 0.08). These results show that increased overall mental health is associated with better QoL, while more emotional distress and worsening patient performance status are associated with poorer QoL. Thus, the CQOLC appears to possess adequate validity, test-retest reliability and internal consistency. Future directions include further evaluation of sensitivity to change and factor analysis to determine the principal health-related QoL domains evaluated by the CQOLC. PMID- 10457739 TI - The adaptations of a quality of life questionnaire for routine use in clinical practice: the Chronic Respiratory Disease Questionnaire in cystic fibrosis. AB - The assessment of quality of life (QoL) is necessary to monitor the course of disease and to assess the effect of new and existing interventions in clinical practice. This will only be achieved if QoL can be measured accurately and routinely. The aim of this study was to demonstrate the methodology involved in the adaptation and shortening of the Chronic Respiratory Disease Questionnaire (CRDQ) in a population of adults with cystic fibrosis (CF). A single interviewer administered the CRDQ to a sample of 45 adult patients (32 males) with CF prior to assessment of spirometric measures of lung function. Those patients whose lung function was stable at the time of study and who could attend for a retest within 14 days were asked to complete the questionnaire at a subsequent visit (n = 10). The mean interval between visits was 7 days (range 5-14 days). Correlations between spirometry and CRDQ dimensions ranged from -0.003 to 0.426. The fatigue, emotion and mastery dimensions showed high internal consistency and adequate construct validity. In the small number of patients suitable for retest, the results indicated that the dimensions exhibited adequate test-retest reliability. In contrast, low internal consistency was demonstrated for the dyspnoea dimension. The fatigue, emotion and mastery dimensions could be reduced, in terms of their number of items, without a substantial loss in explanatory power. This study suggests that QoL measurement can be made convenient and, thus, more easily accessible for routine clinical assessment. PMID- 10457740 TI - Temporal assessment of quality of life of head and neck cancer patients receiving radical radiotherapy. AB - The study was undertaken to evaluate physical, psychological and functional aspects in quality of life (QoL) assessment prospectively in biopsy-proven head and neck cancer patients receiving radical radiotherapy. Fifty male patients were assessed using Karnofsky's Performance Status (KPS), Beck's Depression Inventory (BDI) and the Functional Living Index-Cancer (FLIC). Patient questionnaires were completed before radiotherapy, during 3-4 weeks of radiotherapy and 3 months after radiotherapy. Before the start of radiotherapy, KPS was 91 +/- 10.26, FLIC was 129.98 +/- 33.41 and BDI was 7.10 +/- 4.57. This indicated good performance and functional status with lower depression. In weeks 3-4 of radiotherapy, KPS (71.00 +/- 20.12) and FLIC (81.34 +/- 45.23) decreased, while BDI (16.56 +/- 9.01) increased, indicating impairment in QoL. Three months after radiotherapy, KPS (78.37 +/- 23.0), FLIC (119.51 +/- 43.62) and BDI (9.02 +/- 7.81) improved but were not restored to pre-treatment levels. When patients were scheduled for radical radiotherapy, maximum deterioration in QoL was seen in weeks 3-4. This is the time when maximum supportive care and psychologic counselling is required. PMID- 10457741 TI - The development of an individualized questionnaire measure of perceived impact of diabetes on quality of life: the ADDQoL. AB - The objectives of the study were to design and develop a questionnaire to measure individuals' perceptions of the impact of diabetes on their quality of life (QoL). The design of the ADDQoL (Audit of Diabetes Dependent QoL) was influenced by patient-centred principles underlying the SEIQoL interview method. Respondents rate only personally-applicable life domains, indicating importance and impact of diabetes. Fifty-two out-patients with diabetes and 102 attending diabetes education open days provided data for psychometric analyses. Each of the 13 domain-specific ADDQoL items was relevant and important for substantial numbers of respondents. Factor analysis and Cronbach's alpha coefficient of internal consistency (0.85) supported combination of items into a scale. Insulin-treated patients reported greater impact of diabetes on QoL than table/diet-treated patients. People with microvascular complications showed, as expected, greater diabetes-related impairment of QoL than people without complications. Unlike other QoL measures, the ADDQoL is an individualized questionnaire measure of the impact of diabetes and its treatment on QoL. Preliminary evidence of reliability and validity is established for adults with diabetes. Findings suggest that the ADDQoL will be more sensitive to change and responsive to differences than generic QoL measures. PMID- 10457742 TI - The competing effects of disease states on quality of life of the elderly: the case of urinary symptoms in men. AB - During the period 1993-1994 we conducted a study in Israel on a national-based sample of 960 men to examine the relationships between urinary symptoms and various domains of quality of life (QoL). Regression analyses were performed for each of the eight SF-36 domains, separately for the entire population and for those--without any co-morbidity. The dependent variable was the SF-36 domain scores. The independent variables included age, origin, education, employment and economic status, the degree of disturbance caused by urinary symptoms and the existence of co-morbidities. There was a significant difference between the entire population and the population without co-morbidities. In the entire population we found that severely bothersome urinary symptoms were related to scores on three QoL domains (social function, role-emotional and mental health) but there was no association with physical functioning and general health perceptions. In men without co-morbidity, urinary symptoms were substantially related to physical functioning and general health perceptions. These findings indicate that the relative weight of the impact of a symptom or disease on QoL domains is changed by the presence of other competing factors, such as co morbidities or sociodemographic attributes. PMID- 10457743 TI - A Kiswahili version of the SF-36 Health Survey for use in Tanzania: translation and tests of scaling assumptions. AB - The objective of the study was to translate and adapt the SF-36 Health Survey for use in Tanzania and to test the psychometric properties of the Kiswahili SF-36. A cross-sectional study was conducted as part of a household survey of a representative sample of the adult population of Dar es Salaam, Tanzania. The IQOLA method of forward and backward translation was used to translate the SF-36 into Kiswahili. The translated questionnaire was administered by trained interviewers to 3,802 adults (50% women, mean (SD) age 31 (13) years, 50% married and 60% with primary education). Data quality and psychometric assumptions underlying the scoring of the eight SF-36 scales were evaluated for the entire sample and separately for the least educated subgroup (n = 402), using multitrait scaling analysis. Forward and backward translation procedures resulted in a Kiswahili SF-36 that was considered conceptually equivalent to the US English SF 36. Data quality was excellent: only 1.2% of respondents were excluded because they answered less than half of the items for one or more scales; ninety percent of respondents answered mutually exclusive items consistently. Median item-scale correlations across the eight scales ranged from 0.47 to 0.81 for the entire sample. Median scaling success rates were 100% (range 87.5-100.0). The median internal consistency reliability of the eight scales for the entire sample was 0.81 (range 0.70-0.92). Floor effects were low and ceiling effects were high on five of the eight scales. Results for n = 402 people without formal education did not differ substantially from those of the entire sample. The results of data quality and psychometric tests support the scoring of the eight scales using standard scoring algorithms. The Kiswahili translation of the SF-36 may be useful in estimating the health of people in Dar es Salaam. Evidence for the validity of the SF-36 for use in Tanzania needs to be accumulated. PMID- 10457744 TI - Validation of the Kiswahili version of the SF-36 Health Survey in a representative sample of an urban population in Tanzania. AB - The objective of this study was to assess the validity of a Kiswahili translation of the SF-36 Health Survey (SF-36) among an urban population in Tanzania, using the method of known-groups validation. People were randomly selected from a demographic surveillance system in Dar es Salaam. The representative sample consisted of 3,802 adults (15 years and older). Health status differences were hypothesized among groups, who differed in sex, age, socioeconomic status and self-reported morbidity. Mean SF-36 scale scores were calculated and compared using t-test and ANOVA. Women had significantly lower mean SF-36 scale scores (indicating worse health status) than men on all scales and scores were lower for older people than younger on all domains, as hypothesized. On five of the eight SF-36 scales, means were higher for people of higher socioeconomic status compared to those of lower socioeconomic status. People who reported an illness within the previous 2 weeks scored significantly lower on all scales compared to those who were healthy, as did people who said they had a disability or a chronic condition. PMID- 10457745 TI - A health-related quality of life measure for use in patients with onychomycosis: a validation study. AB - Onychomycosis is a common nail disorder associated with pain, discomfort and varying degrees of physical impairment and loss of dexterity. Psychological and social limitations result from reactions of others to visible impairment. The goal of this research is to validate a questionnaire to measure the impact of toenail onychomycosis on health-related quality of life (HRQoL). One hundred and fifty onychomycosis patients were enrolled in an observational study at eight sites in the US. Attending physicians reported information on clinical status at enrolment. Patients completed a questionnaire covering HRQoL that included general and disease-specific items measuring the impact of onychomycosis on activities and appearance, plus problems and symptoms associated with toenail infection. The subscales of the instrument showed high internal consistency reliability (range = 0.63-0.95). Construct validity reflected the close association of physical functioning scores with onychomycosis impairment. Test Retest reliability was good to excellent for all scales (ICC = 0.52-0.89). Discriminant validity was evidenced by persons who are younger and female reporting worse disease-specific HRQoL. Responsiveness to clinical change was noted for all disease-specific scale scores for improved patients. This instrument has demonstrated reliability, validity and responsiveness for use in observational and clinical studies of toenail onychomycosis patients. Data indicate that onychomycosis patients report significant pain and discomfort reflecting the need for HRQoL measurement. PMID- 10457746 TI - A new measure for assessing the health-related quality of life of patients with vertigo, dizziness or imbalance: the VDI questionnaire. AB - Vertigo, dizziness and imbalance are frequent symptoms among patients in clinical practice. We present the development of a new multipurpose measure specific to patients with these symptoms: the VDI questionnaire. Items of the VDI questionnaire were generated through an international panel of clinicians, in depth interviews with 25 patients and an extensive literature search. Items were reduced by qualitative and quantitative methods. Psychometric properties were tested in 130 patients recruited in primary care general practices, neurologists and ENT out-patient clinics. Patients were clinically evaluated and administered several specific and generic health status instruments: the Balance Scale, the SF 12 Health Survey and the General Health Questionnaire 12 items. The VDI symptoms and the VDI health related quality of life (HRQoL) scales were administered during the visit and some weeks after. Correlation and confirmatory factor analysis were used. Reliability of the VDI was high (0.92 for VDI HRQoL scale and 0.86 for VDI symptoms). VDI scores were more correlated with Balance Scale scores than with mental health indicators. All factor loadings were significantly different from zero and confirmed the expected relations. In the second administration, VDI scores showed a deterioration among those reporting a worsening in their symptoms and an improvement among those reporting feeling better. The data presented suggest that the VDI questionnaire is a reliable, valid and responsive instrument for patients with vertigo, dizziness or imbalance. The VDI questionnaire could be a useful complement of the clinical evaluation of these patients at both group and individual level. PMID- 10457747 TI - Factor structure of the Disability and Impact Profile in patients with multiple sclerosis. AB - The Disability and Impact Profile (DIP) is used for the measurement of quality of life in multiple sclerosis (MS) patients. Data from 211 persons with definite MS from Belgium, Denmark and the Netherlands were used to address three questions. To what extent do the impairment ratings and their complementary weighted scores share common variance? What is the factor structure of the weighted scores? To what extent do disease-related information and information from psychological questionnaires offer concurrent validity for the factors? Correlations between impairment ratings and the weighted item scores were high; for most items the complementary ratings of impact do not supplement information provided by impairment ratings. The DIP weighted scores can be represented to some extent by three independent dimensions: a motor (skeletal movement) or displacement ability factor, a factor relating to sensory-cognitive intactness and a psychological well-being factor. A two-factor solution provides a first factor identical to that of the three-factor solution and a second factor representing psychological well-being. The two factors correlate well with instruments measuring disability and with satisfaction versus emotional distress. Provisional scales based on the factors were constructed. PMID- 10457748 TI - Refining quality of life: validating a multidimensional factor measure in the severe mentally ill. AB - Quality of life measurement has historically been characterized by a focus on physical functionality, great variability in definitions and insufficient attention to psychometric properties of measures. The present study examined four core subscales of the Quality of Life Enjoyment Scale (Q-LES-Q) designed to assess subjective quality of life (i.e. physical health, subjective feelings, leisure activities and social relationships) administered to 151 male and female subjects with severe mental illness admitted to a residential community treatment center or a university psychiatric hospital. The use of factor analysis is a common approach to examining construct validity of instruments through the examination of correlated clusters of item responses. Those sets of highly correlated item responses should identify a construct or dimension of related items (i.e. a factor). Two factor approaches, exploratory (i.e. the maximum number of possible factor is unspecified) and restricted (i.e. the maximum number of factors allowed is specified), were employed to examine construct validity of the four Q-LES-Q subscales. In addition, both orthogonal (i.e. independence between factors is maximized) and oblique (i.e. correlated factors are allowed) rotations (arrangements) of factor structure were also investigated to define subscale validity further. Results supported good construct validity for each subscale with either factor approach, i.e. the four proposed subscales were clearly identifiable in the factors (groupings) of correlated item responses from the sample. In both cases, the orthogonal (independent) rotation produced the simplest structure, i.e. the clearest groupings of items. These results indicate that the Q-LES-Q does appear to measure valid dimensions matching those proposed by the subscales and, thus, may be a useful and reliable tool for clinical applications. PMID- 10457749 TI - Simulation studies: a new method of prospective technology assessment and design. AB - In order to examine the social conditions and effects of a broad usage of telecooperation technology in the future, such as the development of design proposals, PROVET (Projektgruppe Verfassungsvertragliche Technikgestaltung) developed the method of simulation studies. The paper describes the problem of prospective technology design for telecooperation, a method to handle it, the simulation studies and the methological background. It demonstrates three simulation studies, in administration of justice, about office dossiers and for a reachability management. It presents results, particularly about the psychological effects of the digital signature as a technical-organizational measure to guarantee obligations between the cooperation partners. The conclusion discusses this research approach in relation to others. PMID- 10457750 TI - [What is your diagnosis? Aspirate due to dentures lodged in the larynx]. PMID- 10457751 TI - [Human dying]. PMID- 10457752 TI - [Death and dying from the family doctor's perspective]. PMID- 10457753 TI - [Interdisciplinary discussion about the care of the dying--viewpoint of nurses and pastors]. PMID- 10457754 TI - [Interdisciplinary discussion about euthanasia--viewpoint of the clinical physicians]. AB - In western cities more than 80% of deaths occur in the hospital. Thus, we should be familiar with the professional care for dying patients. However, reports of euthanasia in the Netherlands and interviews of patients in other countries demonstrate that medical care for patients with end stage diseases frequently is insufficient. The need for palliative care, which encloses medical, psychological, social and spiritual aspects of the dying becomes apparent. The physical symptoms (e.g. pain) are only one aspect of the suffering of the terminally ill. Following the WHO guidelines for use of analgesic drugs pain control is achieved in the majority of patients. Palliative care may individually tailor the treatment and care to achieve symptom control. Legalization of euthanasia will diminish the interest in practicing palliative care and may also limit the enthusiasm in research in this field as seen in the Netherlands. Data analysis report significant increase of physician-assisted suicide and euthanasia in the Netherlands within five years time from 1990 to 1995 (total: 3.7% to 4.7%, euthanasia: 1.7% to 2.4%). In addition, each year about 1000 patients were not competent at the time euthanasia was performed (euthanasia without request)! Furthermore, a patients illness did not have to be in end stage when he required euthanasia. This information should rise concern about future developments! The public enthusiasm for legalization of euthanasia in Switzerland may reflect the fear of dying and the belief that physicians and other medical professionals are not equipped to adequately care for the dying. Indeed, professional competence of palliative medicine to treat the symptoms of terminally ill patients particularly with cancer has frequently been insufficient. The majority of Swiss dying with assisted suicide (Switzerland belongs to the few countries where assisted suicide is not illegal) in 1996 and more than 80% in the Netherlands dying by euthanasia had cancer. For the terminally ill euthanasia and assisted suicide may seem the only solution. Enhancing education in palliative medicine is a necessary first step to improve the care for the dying patients. PMID- 10457755 TI - [Euthanasia--viewpoint of the practicing physicians]. PMID- 10457756 TI - [The deaths are scandalous as well as ethically questionable]. PMID- 10457757 TI - [What does it mean to live with the understanding that one must take leave of life?]. PMID- 10457758 TI - Self-image and ethnic identification in South Africa. AB - This study examined the relationship between self-image and ethnic identification among 3 South African groups. Participants included random samples of 347 Afrikaans-speaking Whites, 113 English-speaking Whites, and 466 Blacks in urban Gauteng. Positive and negative self-image were extracted using the Rosenberg Self Esteem Scale (M. Rosenberg, 1965). Afrikaans-speaking Whites had the most positive self-image and Blacks the most negative self-image. A positive self image was correlated with stronger ethnic identification among Afrikaans-speaking Whites. The opposite was true for Blacks. This relationship was insignificant among English-speaking Whites. Ambivalence toward ingroup identity was persistently correlated with self-image for all groups. PMID- 10457759 TI - The relationship between collective self-esteem and self-construal in Japan and Canada. AB - The study examined the relationship between various facets of collective self esteem (R. Luhtanen & J. Crocker, 1992) and independent and interdependent self construals (H. R. Markus & S. Kitayama, 1991) among students in Japan and Canada. The results suggested that individuals with highly interdependent self-construals regarded social group memberships as self-defining, regardless of culture. Individuals with independent self-construals judged their groups positively and felt they were worthy members of their groups and that others evaluated their groups positively. Canadians who evaluated their social groups favorably may be members of groups that permitted them to be independent. In contrast, individuals in Japan who judged their groups favorably may be members of groups that promoted interdependence in addition to independence. PMID- 10457760 TI - Psychometric properties of a scale measuring negative attitudes toward overweight individuals. AB - This study examined the psychometric properties of the Anti-fat Attitudes Scale (AFAS), a 5-item instrument developed by the authors to measure negative attitudes toward overweight individuals. A total of 4 studies were conducted among Canadian adolescents (n = 1,452) and university students (n = 424). Results suggested that the AFAS possessed a unidimensional factor structure and satisfactory reliability for both men and women. As predicted, men obtained higher scores than women on the AFAS, and antifat attitudes were positively associated with authoritarianism, homonegativity, and political conservatism. Participants who were overweight had lower scores on the AFAS than participants who were thin or average weight. Responses on the AFAS did not appear to be contaminated by social desirability bias. PMID- 10457761 TI - Longitudinal improvement of self-regulation through practice: building self control strength through repeated exercise. AB - This study examined the results of repeated exercises of self-control in relation to self-regulatory strength over time. A sample of 69 U.S. college students spent 2 weeks doing 1 of 3 self-control exercises: monitoring and improving posture, regulating mood, or monitoring and recording eating. Compared with a no-exercise control group, the participants who performed the self-control exercises showed significant improvement in self-regulatory capacity as measured by quitting faster on a hand-grip exercise task following a thought-suppression exercise. PMID- 10457762 TI - Self-esteem among Arab adolescents in Israel. AB - This study examined the level of global self-esteem of Arab adolescents in Israel and its relationship to perceived academic status and aspirations, interpersonal relationships, community type, and various demographic variables. A group of 1,560 11th- and 12th-grade Israeli-Arab adolescents answered the Rosenberg Self Esteem Scale (M. Rosenberg, 1965). The results revealed significant relationships (a) between global self-esteem and students' evaluations of their scholastic levels, their schools' academic levels, and their plans to take matriculation exams and (b) between self-esteem and family and peer relations. There was a significant relationship between self-esteem and community type. Participants living in cities and villages scored higher than those living in Bedouin townships. There were no significant gender differences or differences among grade levels. PMID- 10457763 TI - The influence of perceived responsibility and personality characteristics on the emotional and behavioral reactions to people with AIDS. AB - The present study examined the effect of a target's personality and responsibility for a misdeed on a perceiver's emotions and behavior by determining the variables' impact on the perceiver's emotional reactions and willingness to help the target. A total of 134 Germans and 171 Americans were given information about the level of responsibility (low, medium, or high) for a person's infection with HIV, 2 variations of information about the target's personality (positive vs. negative), and the transmission mode of the virus (needle vs. unsafe sex). Although B. Weiner's (1995) model of responsibility partially explained the findings, the results showed that the target's personality also contributed to the perceiver's emotional and behavioral reactions. PMID- 10457764 TI - Effect of collective self-esteem on ingroup evaluations. PMID- 10457765 TI - Depression and life satisfaction among northern Irish adults. PMID- 10457766 TI - Negative mood regulation expectancies, coping, and depressive symptoms among American nurses. PMID- 10457767 TI - Sarcomere length-joint angle relationships of seven frog hindlimb muscles. AB - The sarcomere length-joint angle relationship was measured in 7 different muscle joint complexes (n = 43 muscles) of the frog hindlimb (Rana pipiens). Muscles studied included the cruralis, iliacus internus, gastrocnemius, gluteus magnus, gracilis major, semimembranosus and the semitendinosus. Muscle-joint complexes were mounted in a jig and submerged in chilled Ringer's solution. Joints were rotated throughout their range of motion, while sarcomere length was measured by laser diffraction. Muscles were then formalin fixed and architectural properties determined by microdissection of individual muscle fibers. Sarcomere length change per degree of joint rotation (dLs/d theta) ranged from a low of 3.7 nm/degree for the cruralis muscle acting at the knee to a high of 12.5 nm/degree for the semitendinosus muscle acting at the hip. Values for dLs/d theta were significantly different between all muscles (p < 0.001), and dLs/d theta values for muscles acting at the hip were significantly greater than those for muscles acting at the knee (p < 0.005). dLs/d theta was negatively correlated with fiber length, suggesting a balance between fiber length and moment arm in most muscle joint systems. However, many exceptions to this generalization were noted. These data suggest that different muscle-joint systems are 'designed' for differential contribution of muscle force production to the joint torque profile. The low variability of these data also suggests that sarcomere number is tightly regulated in these muscle-joint systems but not simply as a result of the total in vivo muscle excursion. PMID- 10457768 TI - Developmental changes in the distribution of cecal lectin-binding sites of Balb-c mice. AB - The existence of lectin-binding sites was investigated in the cecum of Balb-c mice at seven developmental stages ranging from 18 days post conception (p.c.) to 8 weeks after birth. Nine horseradish-peroxidase-conjugated lectins (concanavalin A, Triticum vulgaris, Dolichus biflorus, Helix pomatia, Arachis hypogaea, Glycine maximus, Lotus tetragonolobus, Ulex europaeus, Limulus polyphemus) were applied to 5- to 7-microns thin paraffin sections of Bouin-fixed tissue. After DAB staining the sections were evaluated by light microscopy. It was shown that each lectin exhibits a unique developmental pattern. The adult binding patterns were established at the age of 3-4 weeks with only minor changes occurring thereafter. Considerable differences in binding patterns occurred not only between lectins of different groups but also between lectins with the same nominal monosaccharide specificity. PMID- 10457769 TI - Influence of continuous electromagnetic fields on the stage, weight and stature of the chick embryo. AB - The influence of continuous electromagnetic fields (0, 181 or 361 Gs/cm2) on the development of chick embryo (n = 144) was studied. Several parameters were determined at days 5, 10 and 15 of incubation: stage (following Hamburger and Hamilton), vertex-coccyx length (size) and weight. At 5 days of incubation, all embryos showed a similar stage. However, at days 10 and 15, the embryos exposed to 181 Gs/cm2 showed a stage significantly superior to that of the others. There were no differences between the exposed embryos and the control ones with regard to weight and stature, except at 15 days when the embryos exposed to 361 Gs/cm2 showed greater weight and stature than those of the controls. PMID- 10457770 TI - Experiments on the induction of antibody dependent macrophage-mediated cellular cytotoxicity in mixed brain cell cultures. AB - These examinations were based on the discussion whether in demyelinating diseases anti-lipid antibody associated with brain macrophages could have a cytotoxic effect on oligodendrocytes. We used mixed brain cell cultures of newborn rats where, among others, both oligodendrocytes and vacuolated macrophage-like cells were found. On these macrophage-like cells, the presence of Fc-receptors was proven. Besides Fc-receptor-dependent phagocytosis, these cells showed an Fc receptor-independent type of phagocytosis. The Fc-receptor-bearing cells moved within the culture and adhered to glass fibers. In the cytoplasm of these cells, unspecific esterase, acid phosphatase and peroxidase could be visualized. The vacuolated cells showed strong autofluorescence, expressed a surface marker found on all types of rat leukocytes and were marked by Griffonia simplicifolia lectin. These results definitely characterized the vacuolised cells as macrophages. We saw globular and pleomorphic macrophages. After incubation of anti-GC serum in a highly diluted solution, significantly more macrophages bound to oligodendrocytes than in the controls. In these cases, we found target cell lysis. It could be shown in vitro that anti-GC serum together with macrophages of neonatal brains can induce a cytotoxic effect on oligodendrocytes. PMID- 10457771 TI - Effect of transforming growth factor-beta on insulin-like growth factor 1- and dexamethasone-induced proliferation and differentiation in primary cultures of pig preadipocytes. AB - The purpose of this study was to examine the effects of a known inhibitor, transforming growth factor-beta1 (TGF-beta1) versus the known stimulators insulin like growth factor-1 (IGF-1) and dexamethasone (DEX) on pig preadipocyte differentiation in serum and serum-free primary cultures. In cultures with serum, preadipocyte and nonpreadipocyte replication was increased (p < 0.02) by IGF-1 and by TGF-beta1 (p < 0.05; p < 0.001). IGF-1 (10 nM) enhanced preadipocyte differentiation (p < 0.05) in serum-supplemented (1% pig serum) cultures, whereas TGF-beta1 (15 pM) reduced preadipocyte differentiation (p < 0.01) in the presence and absence of IGF-1. Furthermore, GPDH (SN-glycerol-3-phosphate dehydrogenase) specific activity (marker that indicates differentiation) was decreased (p < 0.05) by adding TGF-beta1 to serum-free cultures, but TGF-beta1 had little effect in serum-supplemented cultures. DEX significantly enhanced GPDH activity and fat cell cluster number, whereas pretreatment with TGF-beta1 eliminated the DEX enhancement. We have shown for the first time that TGF-beta can decrease (p < 0.01) the cellular secretion of IGF-1 by pig adipose tissue cells and counter the effects of exogenous IGF-1. These studies indicate that TGF-beta1 may not inhibit adipocyte development in the initial growth phase, but may inhibit differentiation and/or hypertrophy (lipid filling) at a later stage of development. PMID- 10457772 TI - Use of LR white resin for post-embedding immunolabelling of brain tissue. AB - The object of this study was to investigate the applicability of the acrylic resin 'LR White' to immunolabelling of various antigenic determinants in aldehydefixed rat CNS tissue. Antibodies were used, which worked well in paraffin sections and therefore were suitable to detect antigens resistant to complete dehydration and heat. Different LR White embedding protocols were employed in order to select the preparation conditions that adequately preserved both the antigenicity and fine structure. Specimens were completely dehydrated with up to 100% ethanol, which was followed by various infiltration times with LR White monomer. Polymerization of the resin was induced by heat, a chemical catalytic procedure (accelerator), or ultraviolet (UV) light. Paraffin, as well as semithin and ultrathin LR White sections were incubated with antibodies reacting to antigens located on the cell surface (stage-specific embryonic antigen-1; SSEA 1), within the plasma membrane (myelin basic protein), in the cytosol (HNK-1, S100 protein), in the cytoskeleton (GFAP, vimentin, neurofilament protein, INT FIL), and in the extracellular matrix (laminin). All of the examined antigens were immunocytochemically detectable in paraffin-embedded material, while the carbohydrate moieties, HNK-1 and SSEA-1, were not immunoreactive in LR White sections. However, in cryostat sections processed for pre-embedding immunoelectron microscopy, the HNK-1 epitope and SSEA-1 were immunolabelled. Polymerization carried out under UV light led to better structural preservation of brain tissue than resin cured with heat or catalyst. The length of prior infiltration with monomer apparently had no effect on tissue preservation. Consequently, UV light-induced polymerization of LR White gives acceptable morphology of brain tissue. However, the use of this acrylic resin is restricted to the detection of some CNS antigens only. PMID- 10457773 TI - Metamorphic changes in the vagal innervation to the alimentary tract in the green frog, Rana clamitans (Ranidae). AB - The alimentary tract of typical anurans shortens extensively at metamorphosis, when the microphagous tadpole transforms into a macrophagous frog. We used methylene blue vital stain and Sihler stain, in conjunction with microdissection, to map the distribution of the gastric branch of the vagus nerve in green frogs (Rana clamitans) before and after metamorphosis. After metamorphosis, terminal divisions of the gastric branch can be observed on the tunica musculosa extending as far as the pylorus. In the tadpole, the gastric branch ends abruptly on the esophagus. The premetamorphic pattern of innervation is consistent with the absence of digestive activity in the foregut before metamorphosis. Preliminary experiments, involving stimulation of the gastric branch of the vagus in the tadpoles, confirm the anatomical observation of the absence of vagal innervation before metamorphosis. PMID- 10457774 TI - [Vessels of the brachial plexus]. AB - The blood supply of brachial plexus is described by means of lightened injected specimens of older human fetus. The following arteries have direct branches to brachial plexus: the aortic arch/brachiocephalica trunk, subclavia a., thyrocervical trunk, ascending cervical a., transverse cervical a., suprascapular a., and the acromial network. Secondary branches leave them that partly anastomose and also return from distal to proximal. The blood supply is luxurious. PMID- 10457776 TI - Anatomy and function of the bursa subacromialis. AB - The ligamentum coracohumerale, as the anterior limit of the bursa subacromialis, runs into the shoulder joint capsule, connecting--broadly based--above the sulcus intertubercularis. It does not, however, originate--as described previously--from the base of the processus coracoideus (viewed frontally), but rather medially from the base, i.e. on the side of the fossa supraspinatus. The sliding mechanism starting at 50 degrees affects the parietal sheet of the bursa subacromialis, whereby the laterally situated section slides under the medially situated section. With an abduction of up to 50 degrees, the lateral section congests in front of the corner of the acromion, sliding from there under the acromion, so that the--hitherto--medial section is located above the section now sliding away beneath it. This sliding mechanism continues on up to 100 degrees. At this point the parietal sheet of the bursa subacromialis lies as follows: The section that had been situated laterally at the beginning of the sliding mechanism now lies caudally to the section that had lain medially at the outset. The bursa subacromialis does not slide fully into the fossa supraspinatus, as in all of the cases observed, it is firmly connected, together with the fascia subdeltoidea, to the corner of the acromion. The visceral sheet does not change in the course of the sliding mechanism as described, as it is connected to the fascia of the supraspinatus muscle--with the exception of a medial stretch of 16 mm. A further finding deals with the course of the muscles of the caput breve of the biceps brachii. Individual muscle fibre components do not connect to the processus coracoideus, but rather run before the tip of the processus coracoideus into the ligamentum coracoacromiale, radiating not only into the ligamentum coracoacromiale, but also--certain components--into the shoulder joint capsule. Through this, the 'aponeurosis tendinis brachii' forms a tendon roof in front of the processus coracoideus that extends to the structures running along the head of the humerus. The muscle fibre components of the caput breve of the m. biceps brachii radiating into the shoulder joint capsule, together with the muscles of the rotator cuff and the ligamentum coracohumerale, keep the shoulder joint capsule tense, thus preventing constriction symptoms. PMID- 10457775 TI - Effects of hypergravity environment on the parathyroid gland of the norepinephrine-treated golden hamster: a stereological study. AB - Effects of 5-gravity environment on the ultrastructure of the parathyroid glands of norepinephrine-treated golden hamsters were studied. In the centrifuged animals treated with norepinephrine, the volume density occupied by the Golgi complexes associated with numerous prosecretory granules was significantly increased compared with that of the control, centrifuged and norepinephrine treated animals, as well as the volume density occupied by the cisternae of the granular endoplasmic reticulum compared with that of the control and centrifuged animals. In addition, in the centrifuged animals treated with norepinephrine, numerous secretory granules were situated close to the plasma membrane. It is suggested that the synthesis and release of secretory granules may be markedly stimulated in the parathyroid glands of the norepinephrine-treated golden hamsters subjected to a hypergravity environment. PMID- 10457778 TI - Specialized cell contacts in the developing nerves of mouse embryos. AB - In mouse embryos, special focal contacts between axons and Schwann cells, axons and fibroblasts as well as Schwann cells and fibroblasts are visible during the outgrowth of nerves. These rather seldom contacts exhibit a uniform structure. Axons and Schwann cells from small finger-like protrusions projecting into coated pits of Schwann cells and fibroblasts. The narrow intercellular space in the contact zone is crossed by fine filaments. PMID- 10457777 TI - Glial fibrillary acidic protein-like immunoreactive ependymal elements in the third ventricle of the rat. A study at different stages of development. AB - Using the peroxidase-antiperoxidase method a study was made of the cells immunoreactive to glial fibrillary acidic protein (GFAP) anti-serum in the ependyma of the third ventricle of the rat at different stages of growth. Most of the ependymal cells of the third ventricle were seen to be unreactive to this protein; however, it was sometimes possible to observe some GFAP-immunoreactive ependymocytes and occasionally other immunoreactive cellular types, such as tanycytes and supraependymal cells. Despite this, the most frequent localization of the elements immunoreactive to the protein adopted the shape of an immunoreactive subependymal band situated parallel to the ventricular wall. As the weights of the animals increased an increase in the elements immunoreactive to this protein could be observed in all the zones considered, there being no differences between the male and female animals. PMID- 10457779 TI - Changes in the lumbosacral angle, sacral inclination and the curvature of the lumbar spine during aging. AB - Measurements of the curvature of the lumbar spine are useful in the investigations of low back pain. It is unclear whether the degree of lumbar lordosis, sacral inclination and lumbosacral angulation are the same for all normal adults. Radiographic studies were carried out on the lumbar spines of subjects aged 9-61 years. Mean and 95% tolerance ranges of the values of lumbar lordosis, lumbosacral angle and sacral inclination for adult age groups up to the sixth decade are given. The results showed that all three parameters varied steadily with age. The pattern of changes differed in males and females. Females had greater angles than males. Sacral inclination appeared to be a more important determinant of the degree of lumbar lordosis. All three parameters showed a tendency to decrease after the sixth decade. The significance of the findings is discussed. PMID- 10457780 TI - Transport of siderotic Kupffer cells to the lung and bronchial excretion of iron in experimental iron-overload. AB - In 3,5,5-trimethylhexanoylferrocene-induced iron overload of rats, three different types of iron-loaded macrophages and derivatives thereof were found in the lungs. On the basis of their localization and of their pattern of iron load it was possible to distinguish: (1) Resident macrophages, showing an alveolar localization and a moderate iron content represented by lysosomal ferritin and haemosiderin. (2) Liver-derived macrophages and giant cells, as well as fragments of them. They showed an exclusive localization in capillaries and alveolar septa, and high concentrations of free ferritin molecules in addition to polymorphous ferritin- and haemosiderin-containing siderosomes. (3) Monocyte-derived intravascular pulmonary macrophages. Initially, they contained iron only as lysosomal aggregates of ferritin and haemosiderin, as a result of phagocytosis of liver-derived macrophageal cell fragments. Later in iron overload, they also showed free ferritin molecules in the cytosol and fused intrapulmonarily to giant cells. The resident as well as the liver-derived siderotic pulmonary macrophages provide a way for iron excretion through the airways. PMID- 10457781 TI - Estradiol influences the LH response to met-enkephalin. An immunocytochemical and morphometric study. AB - In order to test the possible role of estradiol in the response of the LH adenohypophyseal cells to the administration of met-enkephalin in the albino male rat, an immunocytochemical (peroxidase-antiperoxidase), morphometric (cellular and nuclear areas and numerical density) and radioimmunoassay (LH serum levels at the moment of sacrifice) study was carried out. The intraventricular administration of met-enkephalin (150 micrograms in 25 microliters of distilled water) does not produce any changes in the parameters considered. However, when the animals were pretreated with estradiol (chronically, 15 days), met-enkephalin produced a significant decrease in all the parameters considered. PMID- 10457782 TI - Structure and function of matrix components in the cruciate ligaments. An immunohistochemical, electron-microscopic, and immunoelectron-microscopic study. AB - In the present study, the matrix components of 100 cruciate ligaments were analyzed by conventional electron microscopy, immunohistology, morphometry, and immunoelectron microscopy. The anterior (ACL) and the posterior (PCL) cruciate ligaments contained collagen types III, IV, and VI. Several structural glycoproteins, like fibronectin, laminin, entactin, tenascin, and undulin were detected using monoclonal antibodies. Whereas laminin and entactin were higher concentrated in the PCL, type VI collagen was more frequently found in the ACL. The ACL had a critical nourishment in its distal and middle thirds. In all ligament parts the PCL revealed a better vascular supply with strong correlation to type IV collagen expression. The normal matrix of the cruciate ligaments represented a complicated regulatory network of proteins, glycoproteins, elastic systems, and glycosaminoglycans with multiple functional interactions. PMID- 10457783 TI - Capillary supply and fibre area in locomotor muscles of horse and steer--a comparison between histochemistry and electron microscopy. AB - In order to investigate possible differences in variables defining capillary supply of skeletal muscle derived from two methods, the electron-microscopical and the amylase-PAS histochemical methods were applied in a study of horse and steer muscles. Samples from several locomoter muscles were taken at slaughter from one horse and one steer, divided into two and prepared separately for the two techniques. It was found that there was no difference between the two methods in the values for the capillary-to-fibre ratio. Values for mean fibre area, on the other hand, were significantly higher in the histochemically treated samples (16%, p = 0.001) and hence capillary density values lower (12%, p = 0.002). It was concluded that the two methods are comparable in their capacity to visualize capillaries, but preparation of samples for electron microscopy produces an average shrinkage of 16% of muscle fibres. As a consequence capillary density is overestimated accordingly. Discrepancies were larger for steer than horse muscles. PMID- 10457784 TI - [Periosteal and endosteal blood supply of the human fibula and its clinical importance]. AB - The angioarchitecture of the human fibula was investigated by anatomical dissection. The main artery of the periosteal and endosteal blood supply is the arteria fibularis. The proximal part of the facies lateralis is additionally nourished by a well-built branch of the arteria tibialis anterior. The periosteal branches of the arteria fibularis are most voluminous on the dorsal side in the middle of the fibula. Small plates take care of these branches. For an additional revascularization the well-built branch of the arteria tibialis anterior is proposed. The best anatomical region for a vascularized fibular transfer is at an average of 20% up to 75% of the fibular length. PMID- 10457785 TI - Appearance of adipose cells in connective tissue at the implantation site of bone matrix gelatin and bupivacaine injection. AB - Two types of adipose cells were found in the connective tissue on day 7 after bone matrix gelatin (BMG) implantation and an injection of bupivacaine: mature adipose cells with a large lipid droplet (2-140 microns) and immature adipose cells with many small lipid droplets (0.1-2 microns). On day 10 after BMG implantation, typical adipose tissue was observed near the implant. The immature adipose cells had small, spherical mitochondria, glycogen granules and cytoplasmic microvesicles, and they might differentiate from undifferentiated mesenchymal cells in the connective tissue or the peripheral cells around the vessels as a white adipose tissue. These findings suggest that the differentiation of adipose cells in the connective tissue near heterotopic bone formation might be induced not only by mechanical and/or bupivacaine injury, but also by some factor or factors of the BMG. PMID- 10457786 TI - Anatomical study of the blood supply in the human shoulder region. AB - The blood supply of the shoulder muscles, ligaments and joint capsule was investigated by injection of silicone rubber into the arteria subclavia of postmortem cases. We show that the a. suprascapularis forms important anatomoses with other arteries in the region and is of greater importance for the blood supply of the rotator cuff as hitherto believed. We also describe the a. subcoracoidea as a so far unknown branch of the a. axillaris. PMID- 10457787 TI - Evidence of drainage of tungsten particles introduced in the pleural space through the visceral pleura into the lung parenchyma. AB - Studies of pleural clearance of calcium tungstate particles were made in the dog. By using scanning electron microscopy and elemental microanalysis, we show that mesothelial cells of the visceral leaflet of the pleura are also involved in the clearance of particles present in the pleural space. The histological study of lung parenchyma shows many macrophages loaded with tungsten particles, and we conclude that this way may be an important pathway for the transmission of pathologic processes from the pleural space to the lung. PMID- 10457788 TI - Ultrastructure and innervation of water buffalo (Bubalus bubalis) seminal vesicle. AB - The lining epithelium of secretory end pieces and central glandular duct in the seminal vesicle of the water buffalo (Bubalus bubalis) consists of columnar principal and small polymorphous basal cells. A system of intercellular and even intracellular canaliculi enlarges the secretory surface. The most prominent organelle of the columnar principal cells is the granular endoplasmic reticulum, forming large aggregates of parallel lamellae. Using antibodies against the neural cell adhesion molecule L1 and the neural marker protein gene product 9.5 (PGP 9.5), the innervation pattern of the seminal vesicle becomes evident. The muscular layer surrounding the propria contains a dense network of unmyelinated fibers. Thicker bundles traverse the muscular layer to reach the propria. Around glandular secretory tubules and below the epithelial lining of the glandular duct a tightly woven subepithelial plexus is observed which sends short penetrating branches into the basal zone of the epithelium. These intraepithelial nerves are devoid of Schwann cells and basal lamina (naked axons) and are situated within the intercellular spaces between principal and basal cells. Acetylcholinesterase histochemistry with short (1-2 h) incubation times, dopamine-beta-hydroxylase immunohistochemistry and ultrastructural study of transmitter-containing vesicles was performed. The results suggest that muscular contraction in the seminal vesicle is predominantly under the influence of the sympathetic nervous system, whereas secretory epithelial function is regulated by both sympathetic and parasympathetic fibers. PMID- 10457789 TI - 'Synaptic ribbon' modifications in the pineal gland of the albino rat following 24 hours of immobilization. AB - We have studied the ultrastructure of the pinealocyte after 24 h of immobilization. Thirty male albino rats (230 +/- 10 g) were used; the animals were divided into two groups of 15 each. One group was immobilized for 24 h (10.00 h a.m. to 10.00 h a.m.), whereas the other group was used as control. All animals were sacrificed between 10.00 and 14.00 h of the following day. Ultrastructural observations showed an euchromatinic nucleus with numerous giant 'lipid droplets' as well as abundant long 'synaptic ribbons' that were significantly more abundant than in the controls (p < 0.0005). These findings all suggest an increase in pineal activity following 24-hour immobilization. PMID- 10457790 TI - The subcellular localization of the carbohydrate epitope 3-fucosyl-N acetyllactosamine is different in normal and reactive astrocytes. AB - The carbohydrate epitope 3-fucosyl-N-acetyllactosamine (CD15) is involved in cell to-cell recognition processes in various tissues. In the present study the subcellular localization of CD15 was immunocytochemically studied in normal and pathological central nervous system fiber tracts of humans and rats. In normal human white matter of the brain, CD15 immunoreactivity was found on the cell surface of astrocytes and within the cytoplasm of oligodendrocytes. In freshly demyelinated lesions of two human diseases (central pontine myelinolysis and multiple sclerosis) strong cytoplasmic CD15 staining was observed in reactive astrocytes. In normal rats CD15 immunostaining was restricted to the surface of astrocytes. In crush-induced lesions of rat optic nerves, however, astrocytes showed a cytoplasmic localization of CD15, 4 and 6 days after injury. In conclusion, abnormal localization of CD15 in reactive astrocytes may be related to altered functional states of these cells during disease processes. PMID- 10457791 TI - A prospective clinical study of patients with hypernatraemic dehydration. AB - In a clinical prospective 3-year study of 158 children aged 2 weeks to 14 years with hypernatraemic dehydration (serum sodium 150 mmol/l or more), infants predominated (61.4%). The 158 children with hypernatraemia accounted for 13.7% of all children admitted with gastroenteritis over the same period, and significant aetiological factors included the use of artificial feeds, differences between the children with hypernatraemia and those with normo- or hyponatraemia, P < 0.001, P < 0.001, respectively; the use of breast milk, P < 0.001, P < 0.001, respectively; nutritional status, P < 0.001, P < 0.001, respectively; and clinical state of mild to moderate dehydration P < 0.001; P < 0.001, respectively; but not with patients considered severely dehydrated. There was also a significant difference between the presence of neurological features in hyper- and normonatraemic patients P < 0.001; in hyper- and hyponatraemic patients P < 0.05, and in mortality rate between hyper- and normonatraemic patients, P < 0.05 but not between hyper- and hyponatraemic patients. A history of refusal to feed or vomiting was obtained in 41 children (25.9%). The mean serum sodium was 155.5 mmol/l (range 150-189 mmol/l); mean serum urea 7.7 mmol/l (range 1-18.9 mmol/l). Hypernatraemic dehydration remains an important and serious complication of childhood gastroenteritis in our area of study. The use of artificial milk feeds is contributory, and well-nourished babies appear more at risk. We recommend more liberal water intake during gastroenteritis and the public should also be educated on and made more aware of this condition. PMID- 10457792 TI - Amlodipine as monotherapy in hypertensive Africans: clinical efficacy and safety studies. AB - Amlodipine as a monotherapy was evaluated in 20 patients with mild, moderate, and severe hypertension over a 10 week period. After a washout period of 4 weeks, Amlodipine was administered at a dose of 5 mg once daily for 2 weeks and increased to 10 mg once daily if diastolic blood pressure did not fall to below 90 mmHg. At the end of the trial, diastolic blood pressure was reduced to below 90 mmHg in all but four patients. However, these four patients had greater than 20 mmHg reduction in diastolic blood pressure. There was a slight, but insignificant increase in heart rate. Dizziness and weakness occurred in one patient, otherwise, the drug was well tolerated. Laboratory tests, including plasma lipids done at the start and end of the trial, remained unchanged. PMID- 10457793 TI - Biophysical and biochemical analysis of semen in infertile Nigerian males. AB - Biophysical analysis of semen was performed in fifty-eight Nigerian male partners of infertile marriages. Sperm count concentration was significantly higher (P < 0.001) in oligospermics compared to normospermics as expected. However, there was no significant difference in sperm volume or motility percentage between the normospermics and the oligospermics; of course, no sperms were seen in the azoospermics. Biochemical analyses of serum zinc, copper, magnesium, and manganese by atomic absorption spectrophotometry [8] were further correlated in fifty-two patients. There were no statistically significant differences observed in the serum levels of zinc, magnesium, and copper among the normospermics, oligospermics, and azoospermics. The normospermic infertile patients, however, exhibited higher serum manganese when compared with oligospermics and azoospermics (P < 0.001). This finding suggests a potential role for manganese in the evaluation of infertile males. PMID- 10457794 TI - Antimicrobial potentials of Diospyros mespiliformis (Ebenaceae). AB - The petroleum ether, chloroform, methanol, and water extracts of the leaves, stem bark, and root of Diospyros mespiliformis were studied for their antimicrobial activities. The crude extracts showed broad spectrum antimicrobial activities against 9 Gram-positive bacteria, 8 Gram-negative bacteria, and 6 fungal strains. Of the four extracting solvents, chloroform produced extracts with the best antimicrobial activities, while the chloroform extract of the root exhibited the highest antimicrobial activity. Some tetracycline resistant strains of Staphylococcus aureus and gentamicin resistant strains of Pseudomonas aeruginosa were sensitive to some of the extracts tested. Preliminary phytochemical screening revealed the presence of the following metabolites: anthraquinones, tannins, triterpene, saponins, steroids, and sugars and the absence of alkaloids. The antimicrobial activities observed are discussed in relation to the chemical constituents reportedly isolated from several species of this plant and their traditional uses. PMID- 10457795 TI - Determinants of academic and clinical performance in the physiotherapy education programme at the University of Ibadan, Nigeria. AB - A retrospective study was conducted to determine the best predictors of academic and clinical performance in the physiotherapy education programme in the University of Ibadan, Nigeria. Reviewing the records of 94 students enrolled in the programme between 1983 and 1987, multiple and stepwise regression analyses revealed that pre-admission requirements were not significantly related to academic and clinical performance. When all the predictor variables were included in the multiple and stepwise regression analyses, the variance accounted for by the predictor variables was dismally low: 33.8% and 21.3% for academic achievement and clinical performance, respectively. We concluded that the pre admission requirements are not viable predictors of academic and clinical performance in the programme. PMID- 10457796 TI - Effects of acute changes in exercise intensity on basal metabolic rate in fit young men. AB - Basal metabolic rate (BMR) was measured by open circuit indirect calorimetry on 3 consecutive days in 25 highly trained men aged 19-32 years. On day 1, the subjects abstained from all forms of exercise beyond normal sedentary activities. On day 2, they increased the level of exercise beyond their usual level of exertion. Measurements of BMR on day 3 showed a significant but small rise. It is suggested that in the prescription of exercise to patients in weight reduction programmes, bouts of vigorous exercise interspersed by one or more days of relative inactivity, will be more effective than a constant level of exercise. PMID- 10457797 TI - Mechanisms of the blood pressure lowering effect of the calyx extract of Hibiscus sabdariffa in rats. AB - The antihypertensive effect of aqueous extracts of the calyx of Hibiscus sabdariffa (HS) has been investigated in anaesthetized rats. Hibiscus sabdariffa caused a dose-dependent decrease in mean arterial pressure (MAP) of the rats. Sectioning of the right and left vagi nerves did not have a significant effect on the fall in MAP produced by HS. Cholinergic blockade with 0.2 mgkg-1 atropine and histaminergic blockade with 1 mgkg-1 cimetidine and 15 mgkg-1 promethazine significantly attenuated the hypotensive response to HS. Pretreatment of the rats with 20 mgkg-1 HS extract did not have a significant effect on increase in blood pressure induced by bilateral carotid occlusion (48.05 +/- 6.83 mmHg vs 46.53 +/- 7.49 mmHg). The cumulative addition of HS to noradrenaline precontracted aortic rings produced dose-dependent relaxation of the rings. The maximum relaxation response was 86.96 +/- 5.20% and this was observed at the dose of 1.70 mgml-1. These findings suggest that the antihypertensive effect of the extracts of calyx of HS is not mediated through inhibition of the sympathetic nervous system but it could be mediated through acetylcholine-like and histamine-like mechanisms as well as via direct vaso-relaxant effects. PMID- 10457798 TI - Experience with 5-fluoro-uracil and levamisole in the management of advanced rectal carcinoma. AB - Eleven cases of bulky rectal carcinoma occupying the whole of the pelvis are presented. We refer to this advanced rectal carcinoma as "frozen" pelvis. Management of these cases includes performing a divided stoma colostomy, intravenous administration of 5-fluoro-uracil, intraperitoneal instillation of 5 fluoro-uracil when ascites is present, intra-rectal perfusion with 5-fluoro uracil, administration of levamisole, and radiotherapy. We think this regimen improves, to some extent, the quality of life and probably prolongs survival period, although disturbing tenesmus does not disappear completely. This will buttress the argument that abdomino-perineal resection is justified when possible, even if only as a palliative measure to prevent disturbing tenesmus. PMID- 10457799 TI - Evidence that the gastrointestinal tract is involved in glucose homeostasis. AB - Experiments were carried out on fasted anaesthetized dogs. A vein draining a segment of the upper jejunum was cannulated for blood flow measurement and blood samples were obtained for measurement of glucose content of the arterial blood and venous blood from the upper jejunal segment. Glucose uptake was calculated as the product of jejunal blood flow and the difference between arterial and venous glucose levels (A-V glucose). Blood pressure was recorded continuously. The upper jejunum had a resting glucose uptake of 16 +/- 28 mg/min. When adrenaline (1 microgram/kg or 5 micrograms/kg) injection, glucagon (3 micrograms/kg or 8 micrograms/kg) injection or glucose (10 mg/kg/min or 20 mg/kg/min) infusion was given intravenously, glucose uptake increased 600-700%, 700-900%, and 560-600%, respectively at the peak of their responses. These increases in glucose uptake were much greater than increases in blood flow at the peak of responses to adrenaline, glucagon or glucose. Insulin (5 i.mu/kg or 8 i.mu/kg) injection, however, caused a negative glucose uptake of about 400%, i.e., the gut pushed glucose into the blood stream in response to insulin-induced hypoglycaemia. These results indicate that the jejunum plays a modulatory role in glucose homeostasis. PMID- 10457800 TI - The effects of vasoconstrictor on transmission of impulses in the peripheral nerve. AB - The effects of extraneural and intramuscular applications of l-adrenaline on peripheral nerve function were examined. Change in the compound action potential was monitored until a complete conduction block was obtained if it occurred within 100 min. Extraneural application of 0.5 ml l-adrenaline to an isolated segment of the sciatic nerve of the rat in three concentrations of 1:100,000; 1:10,000, and 1:2,000 resulted in complete conduction block within 8 min. However, intramuscular injections of similar concentrations resulted in concentration dependent neural blockades with the highest concentration causing conduction blockade in 27.1 +/- 8.0 min. PMID- 10457801 TI - Prognostic factors in childhood intra-abdominal sepsis. AB - Over a 36-month period, prognostic factors in childhood intra-abdominal abscesses were evaluated in fifty-five Nigerian children (38 boys [69.1%] and 17 girls [30.9%] aged 2 months-15 years (mean 8.9, SD 4.3 years). The mortality rate was 20%. The adverse prognostic factors were: a high fever, a positive blood culture, subphrenic location of abscesses, abscesses associated with a typhoid perforation, post-operative abscesses, presence of organ impairment, and multiple abscesses occurring together in the same patient. The age of the patient, duration of illness before hospitalization as well as the number and type of microorganisms cultured from the abscess did not have any influence on the prognosis in the children in this study. PMID- 10457802 TI - Periodontal disease in homozygous HBSS adolescent Nigerians. AB - The association between sickle cell anaemia (SCA) and periodontal disease was assessed in a prospective comparative study over 6 months using 50 adolescents known SCA patients and 50 adolescent non-SCA subjects. Their ages ranged from 11 to 19 years (mean = 15.25 years). The partial mouth recording system was used to determine the gingival index, plaque index, and probing depths of 6 sampled teeth in the mouth. There was no significant difference between the mean plaque and gingival indices of SCA and the control, but the mean probing depth of the SCA subjects was significantly higher than that of the control, notably amongst the females. However, this was of no clinical significance. The periodontal status of the males in the SCA group was similar to that of their female counterpart whereas the males in the control group have poorer status than the females in the same group. These results suggest that SCA does not lead to increased severity in periodontal disease in adolescent Nigerians. PMID- 10457803 TI - Neonatal tetanus in Port Harcourt. AB - In a previous study on neonatal tetanus (NNT), we provided information on clinico epidemiological data and reported a case-fatality rate of 60.3%. The present report covers a 7-year period, January 1984 to December 1990, and amongst others, focuses on tetanus toxoid vaccine (TT) coverage of pregnant women. The subjects were those with a diagnosis of NNT and other neonatal admissions. Total neonatal admissions, NNT admissions, neonatal deaths, and NNT deaths were: 4,315, 471, 727, and 221 respectively. The overall NNT case-fatality rate was 46.9%. The highest NNT admission rate was in the third quarter of 1984, with a significant decline subsequently except for the increase in 1989. The neonatal tetanus rate began dropping appreciably from 1988 although less than 10% of the mothers in 1990 received two doses of TT. The improper immunization of the mothers during pregnancy with TT is worrying particularly several years into the Expanded Programme on Immunization (EPI), more so as the experience is not unique to Port Harcourt. We suggest intervention at the community level, operational research, and ethnographic studies to determine possible underlying factors to be carried out in all health zones of the country. PMID- 10457804 TI - A review of radiation protection facilities at the University College Hospital, Ibadan, Nigeria. AB - Ionizing-radiation causes health hazards to human beings if exposed to it. Observation of measures allows continued applications of radiation for whatever purpose throughout the world. There are many departments in the University College Hospital that make use of ionizing radiation in some of their services to patients. Radiation protection facilities in the departments were looked into using carefully designed questionnaires. Inadequacies in radiation protection measures were discovered and suggestions for improvements were proffered. PMID- 10457805 TI - Height and weight measurements of Ibadan school children. AB - Height and weight measurements were carried out on three groups of Nigerian primary school children aged 6-12 years, in and around Ibadan. A total of 1,192 children was examined from three social classes as follows: (1) the educated elite group (n = 444); (2) the urban low socio-economic group (n = 366), and (3) the rural village group (n = 382). The school children from the educated elite group had the highest mean height and weight values while the school children from the rural group had the lowest values and the urban low socio-economic came in between the two. In the children of the educated elite class, mean heights and weights were higher than those of the international reference population (though not significantly so) only at ages 6-9 years. Malnutrition as indicated by wasting and stunting was prevalent among both rural and urban low socio-economic school children. The prevalence of wasting was 75.9% among the rural school children, while it was 62.5% among the urban low socio-economic children. The prevalence of stunting was 79.1% among the rural school children, while it was 62.9% among the urban low socio-economic school children. Neither stunting nor wasting was observed among the children of the elite educated group. This study has demonstrated that there has been no change with time in the pattern of differences of height and weight with respect to school children of various social classes. PMID- 10457806 TI - Left ventricular hypertrophy as a risk factor in hypertension. AB - Hypertension is established as one of the major risk factors for congestive heart failure, renal failure, cerebrovascular accident and coronary artery disease. Left ventricular hypertrophy (LVH) is an adaptive response to the increased afterload in hypertension, and therefore serves to normalize wall stress. However, LVH has been established as an independent risk factor for adverse cardiovascular events and death in hypertension. For this reason, considerable attention has been directed towards a better understanding of LVH as a risk factor. As recognition of the risk associated with LVH has grown, investigators have increasingly focused attention on improving methods for the detection of LVH, assessing its effects on cardiac function, defining its relationship with myocardial ischaemia and sudden death, evaluating the role of antihypertensive treatment in the regression of LVH and assessing whether such regression is beneficial in the long term. PMID- 10457807 TI - The effects of high dietary calcium on salt-induced hypertension in rats. AB - The effects of dietary salt-loading and concurrent administration of dietary calcium on water and electrolyte excretion in Sprague-Dawley rats have been studied. The total salt consumed by salt-loaded rats was significantly higher than that of control rats (P < 0.05). The total salt consumed by salt-loaded calcium fed rats was also higher (P < 0.05) than that of control rats but it was slightly lower (P < 0.05) than the consumption by the salt-loaded. Rats fed only high calcium diet had the least salt consumption. The total water intake was also highest (P < 0.05) in the salt-loaded rats when compared with either control rats, salt-loaded-calcium-fed rats or calcium-fed rats. Urinary water excretion was highest (P < 0.05) in salt-loaded-calcium-fed rats followed by salt-loaded rats and calcium-fed rats while the control rats had the least. Similarly, the sodium excretion was highest in salt-loaded-calcium-fed rats followed by salt loaded rats and calcium-fed rats and control rats respectively. Potassium excretion was highest in salt-loaded-calcium-fed rats followed by salt-loaded rats and calcium-fed rats while the control rats had the least excretion of potassium. Serum sodium concentrations were identical in all the groups although in salt-loaded rats and salt-loaded-calcium-fed rats the values were slightly higher than those of controls. However, serum potassium concentration was lowest in salt-loaded rats and calcium-fed rats while the values for salt-loaded-calcium fed rats was not significantly different from that of control. The results of this study suggest that increase in dietary calcium intake reduces the increase in blood pressure resulting from salt-loading and this is accompanied by an increase in water and sodium excretion. PMID- 10457808 TI - Malignant haemangioendothelioma of bone in a HbSC disease patient--a case report. AB - We report here a 35-year-old man with sickle cell disease (SCD), who presented in 1989 with pain in the (R) hip of 7 years duration and swelling of the (R) calf of 3 months duration. Clinical examination revealed a hard tender mass in the (R) calf. Histology of the (R) calf mass revealed haemangioendothelioma (HE), similar to the histology of the (R) iliac bone mass obtained in another institution previously. He was treated with external radiotherapy with the 1.25 megavoltage beam to antero-posterior fields of the (R) hemipelvis and (R) calf, with good response. Chemotherapy was subsequently administered using 6 cycles of VAC regime. The patient remained in remission for 12 months. In 1991 he had lymphoedema of (R) lower limb and received further radiotherapy and chemotherapy after an isotope bone scan had revealed disease activity in the (R) hemipelvis, (R) femur and (L) upper tibia. He responded again with complete regression of the lymphoedema and remained well until April 1993 when the lymphoedema recurred. He died while being evaluated for further treatment. Although there is no evidence to suggest that SCD confers any protection from development of neoplasms, the co existence of SCD with a neoplasm is not common. We consider the occurrence of HE of bone, a rare malignancy, in a HbSC patient worthy of reporting. PMID- 10457809 TI - Oral cavity onlay grafting using foam impregnated with honey:--a case report. AB - The often difficult problem of graft immobilisation in the oral cavity may be surmounted by use of a material which is cheap and readily available. A case in which honey impregnated foam material was used as a stent in oral grafting is described. Foam constitutes an ideal material which is highly recommended. PMID- 10457810 TI - Ectodermal dysplasia with hypodontia in a set of Nigerian twins--a case report. AB - A set of Nigerian male twins with features of ectodermal dysplasia born to apparently normal parents are presented. The main findings in both children were hypodontia, abnormally shaped teeth and hypotrichosis. The general treatment modalities of the condition, especially in a tropical environment, are discussed. PMID- 10457811 TI - Eye care outreach to an underserved rural population in Oyo State. AB - This paper is essentially an initial assessment of work done through a rural community outreach programme based in the ophthalmology department of a large teaching hospital. It evaluates the accessibility, acceptability, and initial effectiveness of the programme by presenting information on selected variables. The evaluation is used to assess the initial achievements of the programme against the planned objectives. PMID- 10457812 TI - Daytime functioning in obstructive sleep apnea patients: exercise tolerance, subjective fatigue, and sleepiness. AB - A sample of 32 obstructive sleep apnea patients (27 males, 5 females) was assessed with overnight polysomnography and the Multiple Sleep Latency Test (MSLT), an objective measure of daytime sleepiness. Patients also participated in a maximal exercise test, which served as an objective indicator of physical fatigue. The Fatigue Severity Scale (FSS) was used as a subjective measure of fatigue. Subjective fatigue ratings were significantly correlated with percent of predicted maximum heart rate achieved during exercise testing, suggesting that self-reported fatigue in apnea patients may refer to reduced physical fitness. FSS scores and exercise testing results were not significantly correlated with the MSLT, indicating that daytime fatigue and daytime sleepiness are independent problems in apnea patients. Participants self-reported a high level of fatigue, and exercise testing revealed decreased physical work capacity among apnea patients, but objective and subjective indicators of fatigue were not significantly correlated with apnea severity. A higher percentage of REM sleep predicted greater work capacity. PMID- 10457813 TI - The treatment of irritable bowel syndrome with hypnotherapy. AB - Previous research from the United Kingdom has shown hypnotherapy to be effective in the treatment of irritable bowel syndrome (IBS). The current study provides a systematic replication of this work in the United States. Six matched pairs of IBS patients were randomly assigned to either a gut-directed hypnotherapy (n = 6) or to a symptom monitoring wait-list control condition (n = 6) in a multiple baseline across subjects design. Those assigned to the control condition were later crossed over to the treatment condition. Subjects were matched on concurrent psychiatric diagnoses, susceptibility to hypnosis, and various demographic features. On a composite measure of primary IBS symptoms, treatment was superior (p = .016) to symptom monitoring. Results from the entire treated sample (n = 11; one subject was removed from analysis) indicate that the individual symptoms of abdominal pain, constipation, and flatulence improved significantly. State and trait anxiety scores were also seen to decrease significantly. Results at the 2-month follow-up point indicated good maintenance of treatment gains. No significant correlation was found between initial susceptibility to hypnosis and treatment gain. A positive relationship was found between the incidence of psychiatric diagnosis and overall level of improvement. PMID- 10457814 TI - Observation of a paradoxical temperature increase during cognitive stress in some chronic pain patients. AB - A total of 224 chronic pain somatoform disorder patients without obvious pathophysiology or psychopathology were found to have colder hands than nonpatients. A paradoxical temperature increase (PTI) in response to a cognitive stressor (mental arithmetic) was noted in a subset of these chronic pain patients. Patients were defined as "PTI" responders if, during cognitive stress, an increase in digital temperature occurred over a prior eyes closed resting condition. It was found that 49.4% of males and 42.6% of females in a total sample of 224 patients demonstrated PTI. The PTI patients had significantly colder hands than non-PTI patients prior to stress. A concurrent SCL measure of sympathetic activation found no difference between the PTI and non-PTI groups either at baseline or during cognitive stress. It appears from this data that PTI is specific to the peripheral vascular system of these patients and may be a marker of psychophysiological dissociation or trauma blocked from consciousness. PMID- 10457815 TI - Neurofeedback combined with training in metacognitive strategies: effectiveness in students with ADD. AB - A review of records was carried out to examine the results obtained when people with Attention Deficit Disorder (ADD) received 40 sessions of training that combined neurofeedback with the teaching of metacognitive strategies. While not a controlled scientific study, the results, including pre- and post-measures, are consistent with previously published research concerning the use of neurofeedback with children. A significant addition is that a description of procedures is included. The 111 subjects, 98 children (age 5 to 17) and 13 adults (ages 18 to 63), attended forty 50-min sessions, usually twice a week. Feedback was contingent on decreasing slow wave activity (usually 4-7 Hz, occasionally 9-11 Hz) and increasing fast wave activity (15-18 Hz for most subjects but initially 13-15 Hz for subjects with impulsivity and hyperactivity). Metacognitive strategies related to academic tasks were taught when the feedback indicated the client was focused. Some clients also received temperature and/or EDR biofeedback during some sessions. Initially, 30 percent of the children were taking stimulant medications (Ritalin), whereas 6 percent were on stimulant medications after 40 sessions. All charts were included where pre- and post-testing results were available for one or more of the following: the Test of Variables of Attention (TOVA, n = 76), Wechsler Intelligence Scales (WISC-R, WISC-III, or WAIS-R, n = 68), Wide Range Achievement Test (WRAT 3, n = 99), and the electroencephalogram assessment (QEEG) providing a ratio of theta (4-8 Hz) to beta (16-20 Hz) activity (n = 66). Significant improvements (p < .001) were found in ADD symptoms (inattention, impulsivity, and variability of response times on the TOVA), in both the ACID pattern and the full-scale scores of the Wechsler Intelligence Scales, and in academic performance on the WRAT 3. The average gain for the full scale IQ equivalent scores was 12 points. A decrease in the EEG ratio of theta/beta was also observed. These data are important because they provide an extension of results from earlier studies (Lubar, Swartwood, Swartwood, & O'Donnell, 1995; Linden, Habib, & Radojevic, 1996). They also demonstrate that systematic data collection in a private educational setting produces helpful information that can be used to monitor students' progress and improve programs. Because this clinical work is not a controlled scientific study, the efficacious treatment components cannot be determined. Nevertheless, the positive outcomes of decreased ADD symptoms plus improved academic and intellectual functioning suggest that the use of neurofeedback plus training in metacognitive strategies is a useful combined intervention for students with ADD. Further controlled research is warranted. PMID- 10457816 TI - Regarding the database for the Peniston alpha-theta EEG biofeedback protocol. AB - Five papers by Peniston and colleagues, which constitute the basic literature for alpha-theta EEG biofeedback treatment for alcoholism and posttraumatic stress disorder, are reviewed. As a result, we raise three questions: (a) Are the samples studied independent? (b) What was the clinical status of the participants prior to treatment? (c) What treatment did the participants actually receive? In seeking answers to these questions we aim to strengthen the database for neurofeedback with specific procedural information so that claims of efficacy can be tested and accepted or rejected on an objective basis. PMID- 10457818 TI - Luciferase reporter gene vectors that lack potential AP-1 sites. PMID- 10457819 TI - Adaptor-aided PCR to identify T-DNA junctions in transgenic plants. PMID- 10457820 TI - Gene dosage by quantitative real-time PCR. PMID- 10457821 TI - High-speed prothrombin G-->A 20210 and methylenetetrahydrofolate reductase C-->T 677 mutation detection using real-time fluorescence PCR and melting curves. PMID- 10457822 TI - Thermal asymmetric interlaced PCR amplification of YAC insert end fragments for chromosome walking in Plasmodium falciparum and other A/T-rich genomes. PMID- 10457823 TI - Hydrogen peroxide improves the efficiency of a peripheral blood PCR assay for diagnosis of human brucellosis. PMID- 10457824 TI - UV light irradiation of plastic reaction tubes inhibits PCR. PMID- 10457825 TI - Specific requirements for PCR amplification of long mitochondrial A+T-rich DNA. PMID- 10457826 TI - Identifying mutations in Drosophila genes by direct sequencing of PCR products. PMID- 10457827 TI - DGGE is more sensitive for the detection of somatic point mutations than direct sequencing. PMID- 10457828 TI - High-resolution, fluorescence-based differential display on a DNA sequencer followed by band excision. PMID- 10457829 TI - DNA analyses from biological material previously used for scanning electron microscopy studies. PMID- 10457830 TI - Chemiluminescence-based method for genotyping Tg.AC responder mice. PMID- 10457831 TI - Accurate mRNA size determination in northern analysis using individual lane size markers. PMID- 10457832 TI - Magnetic separation to concentrate the estrogen receptor from adipose tissue for western analysis. PMID- 10457833 TI - Simple mixing device to reproducibly prepare cationic lipid-DNA complexes (lipoplexes). PMID- 10457834 TI - CO2-enriched atmosphere on the microscope stage. PMID- 10457835 TI - Polyacrylamide gel fixation by drying and gluing to both paper and transparent plastic. PMID- 10457836 TI - A virtual molecular biology teaching laboratory. PMID- 10457837 TI - Screening of transgenic plants by amplification of unknown genomic DNA flanking T DNA. AB - For the screening of transfer DNA (T-DNA) integration in transgenic plant material, we developed a method based on specific amplification of genomic plant DNA flanking T-DNA borders. This approach is possible because the length of the region flanking T-DNA extremity on a restriction fragment is specific to the integration locus. We have modified an adaptor ligation PCR technique developed for amplification of unknown DNA flanking known sequence. The PCR patterns obtained were specific and reproducible for different plants from a given transgenic line. Furthermore, the number of PCR products obtained could be considered a good estimation of the T-DNA copy number. When compared to Southern blot analysis, the PCR results give valuable complementary information about the complexity of the T-DNA integration pattern and also about the integrity of the T DNA borders. We describe the applications of this approach to populations of transgenic Arabidopsis thaliana plants. PMID- 10457838 TI - Nonradioactive labeling of large DNA fragments for genome walking, RFLP and northern blot analysis. AB - In this report, we present a simple nonradioactive labeling procedure for DNA fragments of high specific labeling density that can be used for a variety of applications. The protocol is based on the universal mono-functional platinum reagent for chemical digoxigenin (DIG) labeling of nucleic acids. The labeling protocol was optimized for large DNA templates as complete bacterial artificial chromosomes (BAC). Variations of incubation time and temperature improved the labeling density such that about 30% of the nucleotides were DIG-modified within 30 min. Furthermore, the refined procedure generates in a single-tube reaction and without prior digestion-labeled DNA fragments of 0.5-4.0 kb from a 130-kb template. Hybridization experiments were performed on Southern and northern blots and allowed the detection of single copy genes in 2.5 micrograms genomic DNA from Arabidopsis thaliana, which has a haploid genome size of 0.13 pg (ca. 120 Mb) and medium expressed transcripts from 0.8 microgram poly(A)+ RNA, respectively. The extremely high specific labeling density, the stability and the universal application of the probe generated with the platinum reagent makes this method a useful alternative to classical radioactive nuclei acids labeling techniques. PMID- 10457839 TI - Differentiation of hard-to-type bacterial strains by RNA mismatch cleavage. AB - Many bacteria are difficult to subtype due to high genetic relatedness. In the cases of pathogens of medical or veterinary importance, subtyping is an essential tool of epidemiologists. This report describes a method for molecular subtyping based on the detection of point mutations without DNA sequencing or specialized equipment. The method, known as RNA mismatch cleavage, hybridizes RNA transcripts derived from PCR-amplified DNA, with a control RNA transcript followed by RNase cleavage at point-mutation mismatches. The method was successful in distinguishing all six Brucella species tested and was able to distinguish 11 of the 18 biovars studied. Of the remaining seven biovars (all of which are Brucella abortus strains), three subgroups were identified. The method should be applicable to all hard-to-subtype bacterial strains. PMID- 10457840 TI - Simplified gene-fragment phage display system for epitope mapping. AB - We describe a simple and efficient system for epitope mapping by cloning random gene fragments into a specially designed gIIIp-based phage display vector. DNA encoding the antigen of interest is PCR-amplified and partially digested with DNaseI to generate 50-150-bp-long fragments, which are polished with T4 DNA polymerase and dephosphorylated. These fragments are cloned at the 5' end of the gIII after linearizing the vector with SmaI/SrfI, and the ligation is carried out in the presence of restriction enzyme SrfI. The restriction enzyme in the ligation reaction recuts the self-ligated vector but not the recombinants, since ligation with foreign fragments destroys the enzyme recognition site. Dephosphorylation of inserts prevents their chimerization and ensures ligation of single insert per vector molecule. Thus, using the above strategy, which prevents self-ligation of both the insert and the vector, the overall cloning efficiency and, thereby the library size, is improved more than 10-fold compared to the standard blunt-end, ligation-based methods for making similar libraries. The library is further enriched by a single-step infection of E. coli by phages obtained from primary transformants. This step eliminates all the phages that carry insert that are not in-frame with gIIIp and therefore do not display gIIIp. We have shown the utility of the above system in constructing a glutathione-S transferase (GST) gene-fragment library in phages and identifying the epitope recognized by a monoclonal antibody against GST. PMID- 10457841 TI - High-throughput DNA extraction from old paraffin-embedded biopsies. AB - We have developed a rapid and reliable microporous spin-column-based method for high-throughput extraction of DNA from old paraffin-embedded biopsies. The method was developed specifically for PCRs and molecular genetic studies. De-waxing and DNA extraction from tissues is performed in only one vessel, minimizing pipetting steps and eliminating supernatant/pellet procedures. All steps are performed at 60 degrees, 55 degrees and 95 degrees C minimize deleterious DNase activity. The amount of the extracted DNA was measured spectrophotometrically, and the quality of the extracted DNA and the PCR products was controlled on ethidium bromide stained 3% agarose gel. The presented method for DNA extraction is less laborious and more convenient than previously published methods. It provides means by which extensive, paraffin-embedded material can be subjected to molecular genetic studies. PMID- 10457842 TI - Seven-color, homogeneous detection of six PCR products. AB - We describe an extension of the fluorogenic PCR 5'-nuclease assay, or "Taq-Man" assay. Sequence-specific probes consisted of a novel nonfluorescent quencher, nitrothiazole blue (NTB), at the 3' terminus and six different reporter dyes at the 5' terminus. The six reporters were 6-FAM, dR110, dR6G, dTMR, dROX and JAZ dyes. The seventh color was from aluminum phthalocyanine tetrasulfonate and was utilized as a "passive reference" to calibrate concentration variations. Our test system was a set of three single-nucleotide polymorphisms (SNPs). Each SNP system consisted of two primers and two sequence-specific probes, each labeled with a different reporter dye and NTB. Following PCR, the reactions were diluted with water and measured in a microcuvette on a luminescence spectrometer in synchronous scanning mode. In this method, both the excitation and emission wavelengths were scanned, with a fixed wavelength difference (delta gamma) between excitation and emission wavelengths. The spectral overlap in the set was evaluated by calculation of the condition number of the 7 x 7 matrix (dye fluorescence vs. wavelength). The small value of the condition number (1.5) proved that the cross-talk between the dyes was minimal. SNP analyses of known, synthetic target sequences and genomic DNA were plotted both as normalized, subtracted spectra and as data points in three separate dot plots. PMID- 10457843 TI - Direct observation of nucleocytoplasmic transport by microinjection of GFP-tagged proteins in living cells. AB - We established a straightforward experimental system to investigate directly the requirements for nucleocytoplasmic transport in live cells. For this purpose, substrates were created containing nuclear localization signals (NLS) or nuclear export signals (NES) linked to a chimeric protein composed of the glutathione S transferase (GST) fused to the green fluorescent protein (GFP). The combination of GST/GFP-tagging allowed us to control protein expression in bacteria and to monitor protein purification during chromatography. Following microinjection into somatic cells, nuclear export/import of the highly fluorescent substrates could be observed directly by fluorescence microscopy. This system sets the stage to quantitate, in real time, the kinetics of nuclear import/export in living cells and to evaluate qualitative differences in various NLS/NES signals and pathways. PMID- 10457844 TI - Flow microsphere immunoassay-based method of virus quantitation. AB - A sensitive assay for adenovirus quantitation in vitro was developed using the flow microsphere immunoassay (FMIA) approach. Polystyrene microspheres were covalently coated with purified anti-adenoviral antibodies and incubated with virus-containing samples. After incubation, the samples were stained with DNA specific fluorescent dyes, and microsphere-associated fluorescence was quantitated with a flow cytometer. The adsorption of virus to microspheres was examined under different experimental conditions. The flow cytometric assay was determined to be as accurate in detecting adenovirus as titering on 293 cells. The proposed method can be used to quantify virus in viral stocks and in biological samples. PMID- 10457845 TI - Laser microdissection and laser pressure catapulting for the generation of chromosome-specific paint probes. AB - Chromosome-specific paint probes provide a powerful tool with wide applications in cytogenetic analysis. Here, we present a new approach using UV-laser microbeam microdissection in combination with laser-pressure catapulting, which allows the fast isolation of single chromosomes for the generation of chromosome-specific paint probes. To demonstrate the feasibility of this approach, single chromosomes were collected and amplified with degenerate oligonucleotide-primed PCR, hapten labeled and hybridized onto normal metaphase spreads. Fluorescence in situ hybridization signals revealed specific painting of the respective chromosomes. PMID- 10457846 TI - Strep-tag II for one-step affinity purification of active bHLHzip domain of human c-Myc. AB - The c-Myc protein, the product of the c-myc protooncogene, is a nuclear phosphoprotein with DNA-binding properties when heterodimerized with the Max protein. It contains an amino-terminal transcriptional activation domain and a carboxy-terminal basic helix-loop-helix leucine zipper (bHLHzip) domain that directs heterodimerization and promotes DNA binding. Here, we describe the isolation of the bHLHzip domain of human c-Myc with a technique for efficient single-step purification. Using a C-terminal Strep-tag II affinity peptide and a novel Streptactin-Sepharose matrix, elution is performed under mild conditions by competition with the biotin analog desthiobiotin. No significant influence of the affinity tag on the activity of the bHLHzip domain was observed when the fusion protein was subjected to glutathione S-transferase (GST) pull-down assays for investigating its in vitro-binding properties with GST-Max. The use of the C terminal Strep-tag II was shown to be more suitable for obtaining pure product fractions than use of the N-terminal GST affinity tag. PMID- 10457847 TI - Anterior chamber eye anomalies, redundant skin and syndactyly--a new syndrome associated with breakpoints at 2q37.2 and 7q36.3. AB - We report a 34-year-old female with a de novo balanced reciprocal translocation involving 2q37.2 and 7q36.3. She has a unique combination of multiple congenital malformations that include redundant skin, complete tissue syndactyly of the hands and feet, hirsutism, polycystic ovaries and bilateral anterior chamber eye anomalies. Her son has inherited the unbalanced product (46,XY,der(2) t(2;7)(q37.2;q36.3). He has a similar clinical picture with additional features including complex congenital heart disease, post axial polydactyly, hypotonia and global developmental delay. The breakpoints may indicate the location of the gene(s) responsible for this unique combination of features. PMID- 10457848 TI - Brachydactyly type B: case report and further evidence for clinical heterogeneity. AB - We present a male child with digital and radiographic findings consistent with brachydactyly type B. His left hand had 2-4 syndactyly, shortened first and fifth digits and single palmar and fifth finger flexion creases. The nail on the second finger was hypoplastic. The right hand had a small thumb and curving of the second finger towards the midline. The left foot had a short left hallux with an absent nail, 2-4 syndactyly and an absent toe. His right foot had a short, dorsiflexed hallux with a hypoplastic nail, 2-3 syndactyly and an overlapping toe. Radiographs showed asymmetrical hypoplasia of the middle and terminal phalanges and there were no extra-articular findings to suggest Sorsby syndrome. Brachydactyly type B has classically been described as hypoplasia or absence of the terminal phalanges with absent nails. Although other digital anomalies have previously been described with brachydactyly type B, the absent digit, atypical syndactyly and marked asymmetry in this child make this case a good illustration of the clinical heterogeneity that can be associated with this type of brachdactyly. We provide a brief review of syndromes in which similar digital malformations have been reported. PMID- 10457849 TI - A new syndrome comprising vertebral anomalies and multicystic kidneys. AB - We report a dichorionic twin pregnancy in which both fetuses were affected by a similar pattern of multiple abnormalities. The afrocaribbean parents had a history of infertility, and the pregnancy was conceived using in vitro fertilisation with donor sperm. The features seen in the fetuses do not fit any previously described disorder well. We discuss the differential diagnoses and suggest that this may be an autosomal recessive disorder which has not been previously described. PMID- 10457850 TI - Occipital Horn syndrome in a 2-year-old boy. AB - The clinical presentation of Occipital Horn syndrome, a rare X-linked recessive condition, in a 2-year-old boy is described. This is the youngest patient reported so far. The major clinical, pathophysiological and molecular aspects of this condition are summarized. PMID- 10457851 TI - Oral-facial-digital syndrome with acromelic short stature: a new variant--overlap with Ellis Van Creveld syndrome. AB - The Oral-Facial-Digital syndromes (OFDS) are a heterogeneous group of disorders having common oral, facial, and digital malformations. Here, we report a consanguineous family with a new variety of OFDS associated with acromelic short stature and genu valgum; the features overlapping with Ellis Van Creveld syndrome. One of the sibs has urinary incontinence and growth hormone deficiency, which has not been reported earlier in any type of OFDS in the literature. PMID- 10457852 TI - Spondylo-epi-metaphyseal dysplasia with normal stature: a case followed from infancy to skeletal maturity. AB - Spondylo-epi-metaphyseal dysplasia is a rare skeletal dysplasia showing radiographic abnormalities in the epiphyses and metaphyses of the long and short tubular bones, and in the spine. There are very few reports of a case followed throughout the entire growth period. Here we report the case of a Japanese girl followed from 1 month old to skeletal maturity at 16 years old. She developed progressive kyphoscoliosis at 7 years old, and underwent a surgical correction of the spine at 14 years old. The diagnosis of spondylo-epi-metaphyseal dysplasia was made from examining all the follow-up radiographs showing abnormalities in the spine, long bones and hands involving epiphyses and metaphyses. She had normal stature--a rare finding in this condition. PMID- 10457853 TI - Dominant inheritance of cleft palate with minor abnormalities of hands and feet: a new syndrome? AB - We report a family in which four members of three generations are affected by median cleft palate. The proband and her mother have additional features including bilateral single transverse palmar creases, broad great toes and hypoplastic fifth toenails. Dominant isolated cleft palate has rarely been reported, and there are no previous reports of dominant cleft palate with these associated features. We believe that this represents a previously unreported syndrome, which needs to be considered when assessing recurrence risk for cleft palate. PMID- 10457854 TI - An apparently new acrocraniofacial syndrome with cranial nerve and visceral anomalies. AB - We report details of a neonate with cranial bone dysplasia, broad nasal bridge, microphthalmia, optic and olfactory nerve anomalies, pulmonary segmentation defects, polydactyly, abnormally positioned and shaped thumbs, absent mesentery to the gut and streak gonads. Review of the literature and relevant databases does not identify a likely diagnosis. PMID- 10457855 TI - Radial defects and testicular dysfunction: a new syndrome? AB - We describe two unrelated male patients with radial ray defects and increased serum concentrations of follicle stimulating hormone due to testicular dysfunction. This combination of features has not previously been described. Several mouse genes are known to be involved in preaxial limb development and male fertility, suggesting that the combination of radial ray defects and testicular dysfunction may represent a new syndrome in humans. Abnormalities of spermatogenesis or sperm function may constitute an under-recognized feature of human multiple congenital malformation syndromes. PMID- 10457857 TI - A patient with a de novo t (6;9) and an interstitial duplication of (9)(q21.2q22.1). AB - We report on a 4-year-old child with psychomotor retardation, general hypotonia and only mild dysmorphic features. Her chromosome constitution was 46,XX, t (6;9) (q27;q22.1), dup (9) (q21.2q22.1). This de novo interstitial duplication was confirmed using fluorescence in situ hybridisation (FISH) with band-specific probes. This is the second report of a patient with an interstitial duplication of this region of the long arm of chromosome 9. It is concluded that in a child with an abnormal phenotype and a de novo (apparently) balanced translocation, the possibility of a small duplication or deletion should be considered. PMID- 10457856 TI - Marshall-Smith syndrome: case report of a newborn male and review of the literature. AB - Marshall-Smith syndrome is a rare congenital condition, characterized by advanced bone age, facial anomalies and relative failure to thrive. We report a newborn male with Marshall-Smith syndrome and summarize 21 previously reported cases. We report cerebellar hypoplasia in our patient, which has not been previously reported in subjects with this rare syndrome. This patient's findings broaden the phenotypic spectrum seen in Marshall-Smith syndrome. PMID- 10457858 TI - Aplasia cutis congenita--etiological relationship to antiphospholipid syndrome? AB - A case of aplasia cutis congenita is reported. The mother suffered from antiphospholipid syndrome and a possible etiological relationship between this and aplasia cutis congenita is discussed. PMID- 10457859 TI - Case report on SHORT syndrome. AB - The acronym SHORT was first used by Gorlin et al. (1975) and Sensenbrenner et al. (1975) to define a recognizable pattern of features, consisting of Short Stature, Hyperextensibility of joints and/or inguinal Hernia, Ocular depression, Rieger anomaly, and Teething delay. Other features characteristic of the syndrome included intrauterine growth retardation (IUGR), slow weight gain, frequent illness, triangular face, anteverted ears, telecanthus, deeply set eyes, wide nasal bridge, hypoplastic alae nasi, chin dimple, micrognathia, clinodactyly, partial lipodystrophy, hearing loss, functional heart murmur, delayed bone age, delayed speech, normal intellect, glucose intolerance, and insulinopenic diabetes. To our knowledge 19 cases of SHORT syndrome have been reported (Gorlin et al., 1975; Sensenbrenner et al., 1975; Aarskog et al., 1983; Toriello et al., 1985; Lipson et al., 1989; Schwingshandl et al., 1993; Verge et al., 1994; Bankier et al., 1985; Brodsky et al., 1996; Sorge et al., 1996; Haan and Morris, 1998). We report the twentieth patient diagnosed with SHORT syndrome who presented with growth retardation, sensorineural hearing loss, and minor dysmorphic features, consistent with the phenotype described for this syndrome. PMID- 10457860 TI - Unilateral tibial aplasia, pre-axial polysyndactyly, vertebral anomalies and imperforate anus. AB - Tibial hemimelia is a rare malformation that can be isolated or found with other skeletal abnormalities. We describe a fetus with unilateral tibial aplasia, ipsilateral pre-axial polysyndactyly and femoral hypoplasia, ventriculomegaly, anal atresia and rib and vertebral anomalies. We believe that this combination of malformations has not previously been described. PMID- 10457861 TI - Deletion of chromosome 18 with cardiomyopathy. AB - A female child is described with deletion of chromosome 18 and cardiomyopathy. The clinical features and treatment of the case are described, and the literature of chromosome 18 reviewed. PMID- 10457862 TI - Proteus syndrome: a possible case associated to precocious puberty. AB - We report a boy with possible Proteus syndrome and precocious puberty. This appears to be the first report of this association. PMID- 10457863 TI - An infant with situs inversus totalis, branchial cleft cyst and ectopic kidney: a new combination? PMID- 10457864 TI - Misoprostol used alone for the termination of early pregnancy. A review of the evidence. AB - Increased access to medical methods of abortion could significantly reduce maternal mortality, especially in developing countries. In light of the political and commercial difficulties in distributing the abortifacient mifepristone, the widely studied mifepristone-misoprostol regimen may not be sufficiently available in the near future. Thus, researchers have begun to look for alternative regimens, including regimens using misoprostol alone. This article reviews the current available evidence on the potential of a misoprostol-alone regimen for medical abortion. Although the data are varied and difficult to compare, recent studies indicate that a misoprostol-alone regimen could be safe and effective as a method of medical abortion. Misoprostol is widely marketed around the world for its other indications and is inexpensive, stable in tropical climates, easy to transport, and simple to administer. A misoprostol-alone regimen of medical abortion could thus greatly improve the access to safe medical abortion services by women in developing countries. PMID- 10457865 TI - Early abortion with 800 micrograms of misoprostol by the vaginal route. AB - The objective of this study was to confirm the effectiveness and safety of self administration of misoprostol every 24 h, for abortion up to 9 weeks of gestation. A group of 720 volunteer subjects with gestations from 35 to 63 days received 800 micrograms of vaginal misoprostol every 24 h up to a maximum of three main doses for abortion. Outcome measures assessed included successful abortion (complete abortion without requiring surgery), side effects, decrease in hemoglobin, mean time of vaginal bleeding, and mean time of return of menses. Complete abortion occurred in 644 of 720 (89.4%, 95% CI 87, 92) subjects. The mean decrease in hemoglobin was statistically significant (p = 0.0001). There were 14 subjects with clinically significant decreases in hemoglobin, but only two required transfusions. Vaginal bleeding lasted 6.7 +/- 3.9 days, spotting 8.1 +/- 4 days, and total bleeding 14 +/- 5.3 days. Mean expulsion time was 8.0 +/- 3.4 h. Although mifepristone remains unavailable, given the low price and availability of misoprostol in > 72 countries of the world, this latter drug constitutes an abortion alternative, provided that a minimum clinical network is nearby or accessible. PMID- 10457866 TI - Reasons for contraceptive discontinuation in women 20-39 years old in New Zealand. AB - To estimate the frequency and the medical and nonmedical reasons for discontinuation of oral contraceptive (OC), intrauterine device (IUD), and injectable depot medroxyprogesterone acetate (DMPA) use, data from a cohort of experienced contraceptive users in New Zealand are reported. The current analysis consists of 2469 OC, 2072 IUD, and 1721 DMPA users followed over a period of 5 years. The percentage of women who discontinued the use of the method within 24 months after entry into the cohort were 42%, 44%, and 48%, respectively, for OC, IUD, and DMPA; these differences were not statistically significant. The most common reasons given for discontinuing a contraceptive method, regardless of which method was in use, were the desire to conceive, patient preference, no longer needing contraception, and vasectomy. Among the medical reasons, menorrhagia and intermenstrual bleeding were the reasons for discontinuing use of the method in 1.5 and 1.1 times per 100 women-years among DMPA users and in 1.8 and 4.7 times per 100 women-years among OC users. Pelvic pain and infection were reasons for discontinuing contraceptive method, respectively, 4.4 and 4.3 times per 100 women-years among IUD users. In conclusion, the present study confirms, in this New Zealand population, the high discontinuation rate of contraceptive methods reported elsewhere. In contrast with previous suggestions, in this study, irregular bleeding was not an important medical reason for discontinuation of DMPA use. PMID- 10457867 TI - Eight years with the same IUD. AB - The purpose of this study was to determine the efficacy of the TCu380A (GYNET 380 IUD) in long-term users for periods of > 5 years and to obtain the incidence of Actinomyces as related to length of use and reasons for removal. The report is a retrospective descriptive study of all the clinical cases (n = 370 patients) at the Municipal Family Planning Centre-SERGAS A Coruna, Spain, of women who had a TCu380A IUD inserted during the period 1988-1989 with a September 1997 cutoff date for data analysis. A total of 271 (73.5%) patients had been using the TCu380A IUD for > 5 years with 100% efficacy. The accumulated incidence of Actinomyces was 5.9% (CI 95%: 3.8-8.9). The highest incidence was found between the second and third year of use. There was no increase in the incidence of Actinomyces after 5 years of use. The most common reason for removal of the IUD was the duration of use of 8 years with the same device (32.3%). PMID- 10457869 TI - Timing of onset of contraceptive effectiveness in Norplant implant users. II. Effect on the ovarian function in the first cycle of use. AB - The objective of this study was to time the onset of contraceptive effectiveness in Norplant implant users, when the capsules were inserted beyond the first 7 days of the cycle, based on the immediate effect on the ovarian activity. A total of 42 healthy women requesting Norplant implant contraception were enrolled at clinics in Santo Domingo, Dominican Republic, and in Baltimore, Maryland. Implants were inserted on days 8-13 of the menstrual cycle. Blood samples for estradiol (E2), progesterone (P), luteinizing hormone (LH) (in a subset of 12 women), and levonogestrel (LNG) assay, were taken at 0 h and at 6, 12, 24, 72, and 168 h postinsertion. Ovulation, as defined by P > 2.5 ng/mL, occurred in 40% of subjects. A short lasting, frequently blunted, LH peak occurred within 12 h postinsertion, in all these subjects. The remaining subjects had anovulatory cycles with two distinct E2 profiles: continuously increasing E2 levels to a high mean of 414.3 pg/mL (28%), or no sustained increase in E2 (32%). Most cycles (86%) in which Norplant was inserted with high E2 levels (> 175 pg/mL) were ovulatory, whereas none were ovulatory with low E2 (< 100 pg/mL) at insertion. Based on the endocrine effects of Norplant implant insertion in the midadvanced follicular phase, in which ovulation will either occur within 48 h of insertion or will be impaired, additional contraceptive protection is necessary only for 3 days. PMID- 10457868 TI - Activity of the pituitary-ovarian axis in the pill-free interval during use of low-dose combined oral contraceptives. AB - This study was performed to evaluate pituitary-ovarian recovery in the pill-free interval during use of three low-dose combined oral contraceptives (COC). Either the estrogen component or the progestin component was comparable in the study groups, to evaluate their relative influence. Serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol (E2) levels were measured and follicle number and size estimated by transvaginal sonography daily during the 7 day pill-free interval in 44 healthy volunteers using three different low-dose oral contraceptives. Healthy volunteers were enrolled using 20 micrograms ethinyl estradiol (EE) + 75 micrograms gestodene (GSD) (Harmonet, Wyeth-Lederle; n = 15), 20 micrograms EE + 150 micrograms desogestrel (DSG) (Mercilon, Organon n = 17), or 30 micrograms EE + 150 micrograms DSG (Marvelon, Organon, n = 12) given according to the usual regimen of one tablet daily during 3 weeks and 1 week pill free interval. No ovulations were observed. Pituitary hormones were not statistically significantly different at the beginning of the pill-free interval between the study groups. FSH concentrations were significantly higher at the end of the pill-free interval in the 30 micrograms EE group compared with both 20 micrograms EE groups (7.0 [0.6-12.4] IU/L vs 4.9 [1.4-6.1] IU/L and 4.5 [2.4-7.4] IU/L; p = 0.001). In both 20 micrograms EE groups, a single persistent follicle (24 and 28 mm) was present in one subject. Follicle diameters were statistically significantly smaller at the beginning and at the end of the pill-free period in the 30 micrograms EE group compared with both 20 micrograms EE study groups. Dominant follicles (defined as follicle diameter > or = 10 mm) were observed at the end of the pill-free interval in both 20 micrograms EE groups (in 27% and 18% of women, respectively) but not in the 30 micrograms EE group. Finally, the area under-the-curve for E2 was statistically significantly lower in the 30 micrograms EE group compared with both 20 micrograms EE groups. In conclusion, the EE content rather than the progestin component in the studied COC determined the extent of residual ovarian activity at the beginning of the pill-free interval. Dominant follicles were encountered only in the 20 micrograms EE study groups. PMID- 10457870 TI - Knowledge, attitudes, and practices regarding emergency contraception among nurses and nursing students in two hospitals in Nairobi, Kenya. AB - A cross-sectional descriptive study on knowledge, attitudes, and practice about emergency contraception (EC) was conducted among nurses and nursing students using a self-administered questionnaire. One-hundred-sixty-seven qualified nurses and 63 nursing students completed the questionnaire. Over 95% listed at least one regular contraceptive method but only 2.6% spontaneously listed EC as a contraceptive method, whereas 48% of the respondents had heard of EC. Significantly more nursing students than qualified nurses were familiar with EC. Knowledge about the types of EC, applications, and side effects was poor and 49% of the respondents considered EC as an abortifacient. Of those familiar with EC, 77% approved its use for rape victims and 21% for adolescents and schoolgirls. Only 3.5% of all respondents had personally used EC in the past, 23% of those familiar with EC intend to use it in the future, whereas 53% intend to provide or promote it. The view that EC was abortifacient negatively influenced the decision to use or provide EC in the future. The present findings suggest that the level of knowledge of EC is poor and more information is needed. These findings indicate the potential to popularize emergency contraception in Kenya among nurses and nursing students. PMID- 10457872 TI - Tamoxifen compared to methotrexate when used with misoprostol for abortion. AB - The purpose of this study was to compare tamoxifen to methotrexate, with respect to effectiveness, when followed by misoprostol to induce abortion. In the first phase, 198 women presenting for medical abortion at < 7 weeks gestation were randomized to receive either 40 mg of tamoxifen, followed 2 to 3 days later by 800 micrograms of misoprostol self-administered vaginally or 50 mg/m2 of methotrexate, followed 5 to 7 days later by the same dose of misoprostol. In the second phase, 200 women were randomized to receive 20 mg tamoxifen twice daily for 4 days, followed by 800 micrograms misoprostol or the same regimen of methotrexate and misoprostol as in phase 1. The main outcome measure was success rate determined by the number of women who aborted without surgery. In phase 1, the success rate was higher in the methotrexate group (93.0%) compared to the tamoxifen group (85.7%) (p = 0.045). In the tamoxifen group, nine of 98 women had incomplete abortions with symptoms requiring a surgical aspiration, compared to one of 100 women in the methotrexate group. In phase 2, the success rates were 90.9% in the methotrexate group compared to 84.7% in the tamoxifen group (p = 0.20). The side effects were less in the tamoxifen group in phase 1 but not in phase 2. When tamoxifen is given as a single dose, it is less effective than methotrexate but when it is given 20 mg twice daily for 4 days, there is no significant difference. Tamoxifen does not appear to have any benefits over methotrexate. PMID- 10457871 TI - Contraceptive methods: knowledge sources rated by women and men. AB - People today live amid a welter of information regarding contraception, and health care professionals have been the most prominent sources to which people turn. This study evaluates the relative importance of various information sources and ascertains the position of the physicians among them. Random samples (393 women and 395 men) were drawn from the Finnish population. Response rates were 56% for women and 45% for men. The respondents were asked to estimate the amount of knowledge they had obtained from various sources on a visual analog scale. The three most important sources for women were literature, physicians, and women's journals. For men, literature and the spouse/partner were the most prominent. The youngest age groups--women and men alike--had received more information from the school nurse and the teacher than older groups. Physicians have a central role as information sources. They should be aware of other information sources in the community and should adapt their own work accordingly. PMID- 10457873 TI - Incidence of irregular cycles among Mayan women who reported having regular cycles: implications for fertility awareness methods. AB - Despite the low contraceptive prevalence among the Mayan population of Guatemala, past research has found interest in natural methods. A calendar rhythm method with a simple blanket rule would appear to be preferable to more complicated methods. Under a blanket rule, the number of days of abstinence is predetermined and all couples are instructed to abstain during the same interval of the menstrual cycle. However, regular menstrual cycles may be key to successful practice of a calendar method. A database of 880 cycles of 301 women of the Guatemalan highlands was analyzed to determine the length and regularity of menstrual cycles in this population. PMID- 10457874 TI - The impact of drugs used in anaesthesia on bacteria. PMID- 10457875 TI - Oral tramadol: analgesic efficacy in children following multiple dental extractions. AB - In a randomized double-blind study, 60 children, aged 4-7 years, undergoing dental extractions of six or more teeth under day-case general anaesthesia, were assigned to receive either tramadol drops 1.5 mg kg-1 (n = 31), or placebo (normal saline) (n = 29), 30 min before surgery. In addition, all received anxiolytic pre-medication of oral midazolam 0.5 mg kg-1 (max 7.5 mg) at the same time. No differences were seen in behaviour, respiratory or cardiovascular assessments. In both groups, 93% were drowsy pre-anaesthetic, 3% were asleep but rousable and less than 4% exhibited minor distress. At induction, mild weeping occurred in 9.7% of the tramadol group and 6.9% of the placebo group (P > 0.05). Active awake recovery took 48.8 min, SD 32.6 in the tramadol group and 36.4 min, SD 29.6 in the placebo group (P > 0.05). Post-operative analgesia (paracetamol 120 mg) was given to 19.4% of the tramadol group compared with 82.8% of the placebo group (P < 0.05), after which the Hannalah objective pain scale scores were comparable. Analysis of the Oucher six faces pain scale showed significantly better analgesia in the tramadol group at all time points, the pain score being half that of the placebo group at 60 min and one third from 60 to 120 min (P < 0.05). No adverse respiratory or cardiovascular effects were seen. For children undergoing multiple extractions, 10.7, SD 3.0, effective postextraction analgesia was provided. PMID- 10457876 TI - Light-guided tracheal intubation using a prototype illuminated flexible catheter through the intubating laryngeal mask. AB - We evaluated the efficacy of a newly developed prototype illuminated flexible catheter to facilitate tracheal intubation through the intubating laryngeal mask and compared this light-guided technique with the conventional blind tracheal intubation through the intubating laryngeal mask. The illuminated flexible catheter consists of a completely flexible thin plastic catheter, a bulb attached to its distal end, a 15-mm concentric adapter at its proximal end connected with a battery and a power switch. The device is placed into a silicone tracheal tube in such a way that the bulb protrudes from the distal end of the tracheal tube. One hundred adult patients, ASA I-III, scheduled to undergo propofol/fentanyl/atracurium anaesthesia for elective surgery were studied. All participants underwent a randomized double comparative cross over trial with respect to the tracheal intubation technique through the intubating laryngeal mask. The light guided tracheal intubation was performed as follows; the tracheal tube preloaded with the illuminated flexible catheter was inserted through the intubating laryngeal mask and by observing the glow in the neck was advanced into the trachea. Whenever resistance was felt during insertion, appropriate adjusting manoeuvres were performed. The intubating laryngeal mask was inserted successfully in all patients. The success rate for the blind and light-guided technique was 91% and 100%, respectively (P = 0.003). The mean (+/- SD) duration including appropriate intubating laryngeal mask placement and tracheal intubation, was significantly lower in the light-guided tracheal intubation technique, than with the blind tracheal intubation (31 +/- 8 s vs. 43 +/- 18 s; P < 0.0001). We conclude that the use of an illuminated flexible catheter carries advantages either in optimizing the intubating laryngeal mask position in the laryngopharynx or in achieving a quick and safe light-guided advancement from laryngopharynx into the trachea. PMID- 10457877 TI - Continuous spinal anaesthesia or continuous epidural anaesthesia for post operative pain control after hip replacement? AB - Both continuous spinal anaesthesia and continuous epidural anaesthesia are supposed to provide adequate post-operative pain relief. The purpose of this randomized, prospective study was to compare the quality of analgesia, occurrence of side effects and patient satisfaction between spinal and epidural administration of bupivacaine during the first post-operative 72 h. One hundred and two patients scheduled for hip arthroplasty were randomly assigned to one of two groups: Group 1 received continuous spinal anaesthesia for intra-operative and post-operative management, Group 2 received continuous epidural anaesthesia. Immediately after surgery, the continuous spinal anaesthesia-group received a 1 mL bolus (bupivacaine 0.25%), followed by a continuous infusion of 10 mL over 24 h. The continuous epidural anaesthesia-group received a 10-mL bolus (bupivacaine 0.25%), followed by 2 mL h-1. The level of pain was gauged from a verbal rating score and from a visual analogue scale; the degree of motor blockade was recorded using the Bromage score. In the continuous spinal anaesthesia-group 90.2% reported complete analgesia on the verbal rating scale, but only 21.6% of the continuous epidural anaesthesia-group did. The visual analogue scale scores given by the continuous spinal anaesthesia-group were significantly lower than those of the continuous epidural anaesthesia-group. The percentage of patients with a motor block was significantly higher in the continuous spinal anaesthesia-group on the day of surgery and at the first post-operative day. During the first 24 h, nausea and vomiting occurred more often in the continuous epidural anaesthesia group. The satisfaction was considered excellent in 92.2% of the continuous spinal anaesthesia-group and in 70.6% of the continuous epidural anaesthesia group. It is concluded that continuous spinal anaesthesia and continuous epidural anaesthesia are effective and safe for post-operative pain relief after hip replacement. Compared with continuous epidural anaesthesia, continuous spinal anaesthesia provides faster onset of pain relief, ensures better analgesia and results in more satisfied patients. PMID- 10457878 TI - Comparison of intravenous metoprolol, verapamil and diltiazem on the attenuation of haemodynamic changes associated with tracheal extubation. AB - Changes in heart rate, systolic, diastolic and mean blood pressure were measured after extubation in 60 ASA Grade I and II patients to assess the effects of diltiazem (0.2 mg kg-1), verapamil (0.05 mg kg-1) and metoprolol (0.02 mg kg-1) given as a bolus 2 min before tracheal extubation. All the haemodynamic variables measured increased significantly after extubation in the control and diltiazem groups when compared with the base-line recordings (P < 0.05). Metoprolol effectively blocked the increases in heart rate after extubation and the increase in blood pressure in this group was less when compared with the control group (P < 0.05). Verapamil alleviated the increase in both heart rate and blood pressure. However, profound hypotension and bradycardia requiring therapy, occurred in the verapamil group. For this reason, careful observation is necessary when using verapamil and the routine use of this drug in patients with coronary artery disease requires further studies. PMID- 10457879 TI - Infusion pump performance in an MR environment. AB - Concerns about life support equipment accompanying the critically ill patient have to date made magnetic resonance imaging (MRI) studies of this patient group the exception. We present here a series of tests performed on an IVAC P3000 infusion pump to investigate its suitability for the magnetic resonance imaging environment. We investigate safety, pump performance and image quality issues. The pump was housed at the end of the patient couch to prevent motion towards the scanner. Gravimetric tests found the pump to work within acceptable parameters at a static field of 10 mT. Image interference issues were addressed. PMID- 10457880 TI - Tramadol suppositories are less suitable for post-operative pain relief than rectal acetaminophen/codeine. AB - The suitability of tramadol suppositories for inclusion in our hospital formulary for the treatment of mild to moderate post-operative pain was evaluated. In an open randomized trial, rectal tramadol was compared with our standard treatment acetaminophen/codeine suppositories. We expected tramadol to be equally effective as our current standard but with fewer side effects. Forty patients were treated with either tramadol suppositories 100 mg 6 hourly (qds) or acetaminophen/codeine suppositories 1000/20 mg qds. Patients were comparable with regard to demographic data and type of surgery and anaesthesia. Post-operative pain was scored with the aid of a Visual Analogue Scale before each drug administration, at rest and during movement. Side effects, notably nausea and vomiting, were recorded by interviewing the patients and by inspecting the nursing report. There was no difference in pain scores between the two groups. The incidence of nausea and vomiting was significantly higher in the tramadol-treated (84%) than in the acetaminophen/codeine treated group (31%). The relative risk of experiencing an episode of nausea under treatment with tramadol was 2.7 (95% confidence interval: 1.3-5.3; P = 0.0001) as compared with acetaminophen/codeine. We conclude that for acute treatment of mild to moderate post-operative pain frequent nausea and vomiting makes tramadol suppositories less suitable than acetaminophen/codeine. PMID- 10457881 TI - A simple in vitro method for the evaluation of an ultrasonic nebulizer for drug delivery to intubated, ventilated patients and the effect of nebulizer and ventilator settings on the uptake of fluid from the nebulizer chamber. AB - Little is known about the performance of ultrasonic nebulizers during different ventilation patterns when these nebulizers are used to deliver drugs to intubated, ventilated patients. A method that enables the performance of an ultrasonic nebulizer to be evaluated is described. We used an in vitro model to examine the performance of the DeVilbiss Ultra-Neb 2000 ultrasonic nebulizer under positive pressure ventilation. Performance was measured at different rates of nebulization and under changing conditions of positive end-expiratory pressure (PEEP), inspiratory flow rate, inspiratory time and minute volume. The volume of saline nebulized was unchanged by variations in positive end expiratory pressure from 0 to 5 cm to 10 cm H2O, in minute ventilation and in inspiratory flow rate. An increase in the inspiratory time resulted in an increase in the volume of saline nebulized and this volume was greater as the power setting of the nebulizer was increased. We conclude that ultrasonic nebulizers may be affected by different patterns of ventilation and that this simple in vitro assessment of nebulizer function in an intensive care setting may be of value prior to nebulizer use. PMID- 10457882 TI - Uneventful propofol anaesthesia in a patient with acute intermittent porphyria. AB - A patient with acute intermittent porphyria was anaesthetized with propofol as part of general anaesthesia for cholecystectomy. Post-operatively, no clinical sequelae resulted and urinary porphyrins did not exceed pre-operative values. PMID- 10457883 TI - Recurarization in the recovery room. AB - A case of recurarization in the recovery room is reported. Accumulation of atracurium in the intravenous line led to recurarization after flushing the line in the recovery room. A respiratory arrest with severe desaturation and bradycardia occurred. Circumstances leading to this event and the mechanisms enabling a neuromuscular blockade to occur, following the administration of a small dose of relaxant, are discussed. PMID- 10457884 TI - Transient left vocal cord paralysis during laparoscopic surgery for an oesophageal hiatus hernia. AB - A 45-year-old male, with symptoms of many years standing of gastro-oesophageal reflux disease, was subjected, under general anaesthesia, to laparoscopic fundoplication. Tracheal intubation yielded no problems but great difficulties were encountered during tube insertion into the oesophagus. After surgery, aphonia developed. Laryngological examination demonstrated paralysis of the left vocal cord. Voice strength returned to the pre-operative status after 3 months, and laryngological examination confirmed normal mobility of both cords. The possible cause of the complication was damage to the left recurrent laryngeal nerve which occurred during insertion of the tube into the oesophagus. Gastro oesophageal reflux disease causing 'acid laryngitis' can create conditions favouring this type of complication. PMID- 10457885 TI - An outbreak of food-borne botulism. PMID- 10457886 TI - Analysis of the polymorphism of the tumour necrosis factor (TNF) gene and promoter and of circulating TNF-alpha levels in heart-transplant patients suffering or not suffering from severe rejection. AB - Plasma TNF-alpha levels are generally higher in heart-graft patients who experience a rejection episode than in those who do not. Because the TNF gene and its promoter are polymorphic, we studied the relationships between genetic variability at the TNF locus, the occurrence of graft rejection and TNF-alpha plasma levels in 62 heart-transplant patients in order to investigate inter individual differences in plasma TNF-alpha levels after allogeneic stimulation. TNF-alpha was immunoenzymatically measured in blood specimens collected on the same day as endomyocardial biopsy. After PCR amplification of DNA, NcoI and AspHI polymorphisms were characterized by their restriction profiles, TNFa microsatellites by electrophoretic separation on acrylamide and the promoter region by sequencing. Plasma levels and molecular genetic results were compared to the grade of heart graft rejection established according to pathological criteria. In our study, allograft rejection was associated neither with NcoI or AspHI polymorphism nor with nucleotide changes in the TNF-A promote. We observed low TNF-alpha levels in n1/n1 homozygous patients and in subjects with G-->A at position--308 of the promoter sequence. Concerning the polymorphism of the TNFa microsatellite, our results might suggest an association with graft rejection but we have to be very careful in drawing conclusions because of the small size of the sample. PMID- 10457887 TI - Mutation screening of the interferon-gamma gene as a candidate gene for multiple sclerosis. AB - Interferon-gamma (IFN-gamma) plays a crucial role in the regulation of the immune response. Alterations in IFN-gamma production have been found in several diseases including multiple sclerosis (MS). Such alterations could be caused by the action of different factors on cytokine production, or, theoretically, by mutations in the gene. We screened the IFN-gamma gene promoter and part of the first intron, known to contain a c-Rel specific enhancer, for possible mutations by sequencing. We found a C to T substitution in the IFN-gamma promoter at position -333. Screening for this mutation by sequence-specific PCR in 214 MS patients and 164 controls identified two patients, both heterozygous, but no controls with this mutation. No mutations were found in the first intron. The interferon-gamma gene is highly conserved and changes in IFN-gamma expression are probably due to the influence of regulatory factors on gene transcription, rather than gene polymorphisms. PMID- 10457888 TI - The Bg/II polymorphism of the human prolactin gene lies within intron C and can be detected by PCR/RFLP. AB - Prolactin has been shown to be active as an immunomodulatory hormone and is therefore of potential importance in disease progression and development. Any polymorphism in the gene and regulatory sequences may prove useful for disease association studies. A Bg/II polymorphism has been previously detected within the prolactin gene region. We have mapped this polymorphism to intron C and detected the base mutation that causes it. We have also developed a PCR-RFLP method to genotype individuals. PMID- 10457889 TI - Genotyping for human platelet-specific antigens HPA-1, -2, -3, -4 and -5 in the Slovenian population reveals a slightly increased frequency of HPA-1b and HPA-2b as compared to other European populations. AB - Typing of human platelet alloantigens (HPA) is necessary in various clinical situations. The purpose of this study was to type a random sample of the Slovenian population for HPA alleles, in order to obtain genetic population data. A total of 152 unrelated Slovenian blood donors were genotyped for HPA-1, -2, -3, -4 and -5 alleles using a simple method that enables simultaneous and complete determination of HPA genotypes. Ten different polymerase chain reactions employing sequence-specific priming (PCR-SSP), which worked in identical cycling conditions, were used. The allele frequencies were 0.809 for HPA-1a, 0.191 for HPA-1b, 0.891 for HPA-2a, 0.109 for HPA-2b, 0.591 for HPA-3a, 0.407 for HPA-3b, 0.997 for HPA-4a, 0.00 for HPA-4b, 0.934 for HPA-5a and 0.066 for HPA-5b. When compared to results of studies of various other Caucasian populations, our population displayed a slightly but not significantly higher proportion of the HPA-1b and 2b alleles. PMID- 10457890 TI - HLA-G and classical HLA class I transcripts in various components of the adult human eye. AB - HLA-G transcripts have previously been detected in the ocular cell line HS738, in foetal eyes and in global ocular extract. Using RT-PCR and hybridization, we searched for HLA-G and classical HLA class I transcripts in the corneas of eye donors (n = 6) and in various components of eyes collected at time of surgery: lens (n = 3), iris (n = 2), vitreous body (n = 2) and retina (n = 2). Whereas HLA class I mRNA was abundant in the ocular tissues of all donors, we could not find HLA-G transcript in any ocular tissues obtained from subjects in vivo. The absence of HLA-G in adult eye cornea limits its interest as a possible target in corneal transplantation. Classical HLA class I transcripts were abundant in the cornea and other components of the eye. These results highlight the role of classical HLA class I antigen in eye immunity and corneal transplantation. PMID- 10457891 TI - Heterogeneity of HLA-DR2 haplotypes in Caucasoid Americans, African Americans, Chinese Americans, Native Americans and Xiamen Chinese. AB - HLA-DR, -DQ specificities were determined by PCR amplification with SSOP in 4560 individuals: Caucasoid Americans (CA), African Americans (AA), Chinese Americans (ChA), Native Americans (NA) and Xiamen Chinese (XC). DR2 subtypes were compared amongst the five ethnic populations. The DRB1*1501-DRB5*0101 haplotype was found to be the most frequent in all populations except African Americans, in which DRB1*1503-DRB5*0101 was the predominant haplotype, accounting for 65% of DR2 subtypes. In contrast to Caucasoid Americans, the DRB1*1602 is strongly associated with the DRB5*0101 allele in Chinese populations. The presence of DRB5*0203 and DRB1*1602-DRB5*0101 haplotypes in Chinese populations, especially in Xiamen Chinese, suggests that various DR2 haplotypes may be generated via multiple gene conversion events together with point mutations and reciprocal recombination. The strong DR and DQ associations are found in DRB1*1501/DQB1*0602 (66.22%) for CA, DRB1*1503/DQB1*0602 (56.58%) for AA, DRB1*1501/DQB1*0602 (30.20%) and DRB1*1602/DQB1*0502 (15.76%) for ChA, DRB1* 1501/DQB1*0602 (41.55%) and DRB1*1602/DQB1*0301 (40.25%) for NA, and DRB1*1501/DQB1*0602 (30.26%) and DRB1*1602/DQB1*0502 (25.81%) for XC. PMID- 10457892 TI - HLA-DPB1 alleles in house dust mite allergic patients. AB - Associations were sought between the presence of allergic sensitivity to Der p 1, a major allergen from the house dust mite, and the HLA-DPB1 genotype. Whilst allergic patients did not differ from controls in DPB1 allelic distribution, there was a correlation of DPB1*11011 with strongly reactive T-cell proliferative responses to Der p 1 and high titre specific IgE to Dermatophagoides pteronyssinus. PMID- 10457893 TI - Antigenic stimulation in T-cell cultures in cardiomyopathies: differences in cytokine profiles. AB - Immune-mediated mechanisms are involved in the pathogenesis of cardiomyopathies. In this study, we investigate which pattern of immune response (Th1 or Th2) lies behind these diseases by analysing the basic cytokines secreted from PHA-cultured T lymphocytes and determining what differences, if any, exist between dilated cardiomyopathy (DMC) and hypertrophic cardiomyopathy (HCM). Two groups of patients were studied: 10 patients with DCM and 10 patients with HCM. Age- and sex-matched healthy individuals were used as controls. PHA-cultured T lymphocytes in the presence or absence of different myocardial antigen (MA) concentrations were measured. Interleukine-2 (IL-2), Interleukine-6 (IL-6) and Interferon-gamma (IFN-gamma) levels were measured in culture supernatants by an ELISA method. At the same time, delayed-type hyperactivity (DTH) against the same antigenic preparation was measured by the leukocyte migration inhibitory index technique. Patients were subdivided into DTH-positive and DTH-negative and re-examined for IL-2 cytokine expression. IL-6 levels were found to increase both in the presence and in the absence of MA in the patient groups compared to the controls. IL-2 levels were decreased in both groups, in an antigen dose-related manner. Anergic patients showed a further reduction in IL-2 levels for both groups of patients. IFN-gamma remained unaffected in the patient groups. Almost half of the patients exhibited anergy to the DTH reaction against MA. We conclude that, upon antigenic stimulation, the initially mounted immune response (increased IL-6) is somehow blocked/switched off in patients, resulting in an immunologic tolerance/unresponsiveness to MA (IL-2 decreased, IFN-gamma unchanged). Finally, increased IL-6 could lead to a perpetuation of immunologic injury through the release of oxygen-free radicals with a cytotoxic effect on the myocardium. We hypothesize an antigen-related, defective macrophage-Th1 cell reaction, which accounts for the differences in the IL-2 profile between the DCM and HCM groups, that might cause local immune responses to lead to immunosuppression (immune tolerance effect), thus contributing to the pathogenesis of cardiomyopathies. PMID- 10457894 TI - Susceptibility to coeliac disease in Tunisian children and GM immunoglobulin allotypes. AB - Coeliac disease is a malabsorption disorder of the small intestine resulting from ingestion of gluten. The immunogenetic component is clearly demonstrated by the association of the disease with human leukocyte antigens (HLA). Among other candidate genes are the GM allotypes, which are the markers of the constant parts of heavy chains of the subclasses IgG1, IgG2 and IgG3. GM immunoglobulin allotypes have been analysed in 131 unrelated Tunisian children with coeliac disease. All patients and their parents were tested for G1M(1, 2, 3, 17), G2M(23) and G3M(5, 6, 10, 11, 13, 14, 15, 16, 21, 24, 28) by the classical haemagglutination method. Genotypes and haplotypes were deduced from phenotypes in patients and their parents. Transmission disequilibrium tests were performed in 79 informative families. The GM*3;..;5* haplotype was transmitted more often (23) than not (8) by heterozygous parents (chi 2 = 7.26; P = 0.007). This difference remained significant after correction for multiple testing. This study provides evidence for association and linkage between GM and coeliac disease. It suggests that GM or genes close to GM play a role in the development of the disease. PMID- 10457895 TI - Nationwide collaborative study of HLA class II associations with distinct types of juvenile chronic arthritis (JCA) in Greece. AB - The aim of this study was to investigate the association of different groups and subgroups of juvenile chronic arthritis (JCA) with HLA class II (DR, DP, DQ) alleles and/or haplotypes. Groups and subgroups were mainly distinguished on the basis of the type of onset, the course and complications of the disease, and some predefined disease markers according to the criteria proposed by the ILAR Standing Committee (Chile, 1994). On the basis of these criteria the following five JCA groups and their subgroups were included in the study: (1) define systemic onset (n = 25) and systemic progressing to persistent arthritis (n = 14); (2) JCA of oligoarthritis onset (O-JCA, n = 124) and of oligoarthritis onset and course (n = 98), O-JCA of early (< 6 years) or late (> 6 years) onset (EOO JCA n = 71 and LOO-JCA n = 44), O-JCA with ANA positive (n = 69) or negative (n = 55) and O-JCA progressing to extended arthritis (n = 22); (3) JCA of polyarthritis onset (P-JCA) with rheumatic factor (RF) negative (n = 29), and P JCA RF negative with antinuclear antibodies (ANA) positive (n = 13) or negative (n = 16); (4) JCA complicated with chronic anterior uveitis (CAU, n = 32); (5) juvenile psoriatic arthritis (n = 20). To assess the HLA allele frequencies in the above 223 Greek children with JCA, these frequencies were compared to those of 98 age-matched and 250 adult controls. The main findings were the following. A common HLA-DRB1* allele was not involved in the JCA groups and subgroups studied; on the other hand, the DQA1*0501 allele was found to be associated with different JCA groups/subgroups (O-JCA, P-JCA RF-negative ANA-positive, JCA with CAU), probably suggesting a closer relationship of this locus with the immunogenetic background of JCA. The DPB1*0201 allele was associated with the development of either EOO-JCA or CAU. Susceptibility to CAU was stronger when the DPB1*0201 was combined with the presence of DRB1*13. Another allele, DQB1*0301, was also associated with O-JCA and CAU. Finally, no specific HLA class II allele was found to be related to the presence of ANA or psoriatic lesions or to the severity of the arthritis. Our findings suggest that the wide clinical and laboratory spectrum of JCA is associated with an immunogenetic background that is linked with HLA alleles of more than one locus. Some of them, such as the DPB1*0201 allele, confer susceptibility to certain clinical onsets and courses or complications of the disease. The rapidly advancing techniques of typing of DNA profiles may lead to more definite conclusions. PMID- 10457896 TI - A complex pattern of microsatellite polymorphism within the bovine NRAMP1 gene. PMID- 10457897 TI - Nomenclature for factors of the HLA system, update January 1999. PMID- 10457898 TI - Nomenclature for factors of the HLA system, update February 1999. PMID- 10457899 TI - Nomenclature for factors of the HLA system, update March 1999. PMID- 10457900 TI - Measuring respiration of cultured cell with oxygen electrode as a metabolic indicator for drug screening. AB - New trend in methods for assessing pharmacological action to bacteria and cell is to measure their metabolic activities induced, while the conventional methods used population growth. We focused on respiration volume as an indicator of cell metabolism, and developed inexpensive disposable oxygen electrode sensor and multi-channel dissolved oxygen meters (DOX-10 and DOX-96KB). Using these instruments, cytotoxicity was measured for 48 hrs and the method showed superior features to conventional methods in its handiness of one step assay, and excellent adaptability to automated systems. Total usability of this oxygen electrode method is being evaluated in bacterial drug susceptibility test, anticancer drug susceptibility test, and alternatives to animal experiment. PMID- 10457901 TI - [Angiogenesis--vascular endothelial growth factor and its receptors]. AB - Angiogenesis is a crucial biological process not only in the formation of cardiovascular system and organization of tissues in embryo but also in a variety of diseases including solid tumor growth and diabetic retinopathy. Several protein factors crucial for regulation of angiogenesis have recently been identified. Among these factors, VEGF is considered to be the most important regulator for vascular endothelial cell growth and differentiation both in physiological and pathological conditions. Relationship between VEGF and other regulatory factors such as Angiopoietins should be elucidated to further understand the dynamic process of angiogenesis. PMID- 10457902 TI - Implication of vascular endothelial growth factor in the development and metastasis of human cancers. AB - Vascular endothelial growth factor (VEGF) is a most potent angiogenic molecule. In this article, we demonstrated that VEGF is participated in the tumor angiogenesis of hepatocellular carcinoma, esophageal cancer, and pancreatic cancer. Furthermore, we revealed that VEGF is one of the molecules which are responsible for metastasis and prognosis in esophageal cancer and colon cancer. Although the mechanism on the induction of VEGF gene is still unclear in human cancer tissue, we obtained the informative evidence indicating that p53 mutation is involved in VEGF expression of esophageal cancer. Our experimental study with stable transfectant of VEGF gene provided the confirmative results showing that VEGF gene induces neovascularization in and around tumor and that VEGF augment metastastic potential by accelerating proliferative activity after reaching the target organ. PMID- 10457903 TI - [Angiogenesis in vitro]. AB - A quantitative angiogenesis in vitro was investigated by culturing bovine carotid artery endothelial cells between two layers of type I collagen gel. Cells become organized into tube-like structures within few days. Ultrastructurally, tubular structures were composed of one to several endothelial cells containing pinocytotic vesicles and cytoplasmic projections, and linked by junctional complexes. A basal lamina-like structure surrounded the abluminal surface. Glucose, insulin, insulin-like growth factor I at pathophysiological high concentrations significantly stimulated tube-forming activity of endothelial cells by stimulating cell migration. PMID- 10457905 TI - [Smooth muscle myosin of SM1 and SM2 isoforms expressing human neuroblastoma cell line of MP-N-MS]. AB - Human neuroblastoma (NB) cell lines have at least three morphological appearance of neuroblastic (N-type), substrate-adhessive (S-type) and intermediate(I) cells. Our previous study revealed S-type cells expressed alpha-smooth muscle actin, desmin and/or basic-calponin, indicating the plausible smooth muscle cell characteristics of S-type cells. In this study, a new human NB cell line, MP-N MS, was established from bone marrow metastasis of a one year and six-month old girl with advanced NB, originating from right adrenal gland. Morphology of this cell line is composed of S-type cells. MP-N-MS was identified as a NB cell line by surface membrane antigen analysis and MYCN gene amplification. EWS-FLI1 and EWS-ERG chimeric products, observed in Ewing family tumors, were not detected by RT-PCR (reverse transcriptase-polymerase chain reaction). In cytoskeletal protein analysis, alpha-smooth muscle actin and basic calponin of smooth muscle cell markers were detected. Furthermore, smooth myosin of SM1 isoform was identified in MP-N-MS cell line by immunofluorescence, Western blot and RT-PCR, whereas smooth myosin of SM2 was detected by RT-PCR. MP-N-MS is the first cell line, showing SM1 and SM2 isoforms. The presence of smooth muscle myosin of SM1 and SM2 isoforms in MP-N-MS demonstrated the mature smooth muscle phenotype of this NB cell line, and the ability of NB cells to differentiate into smooth muscle cell. PMID- 10457904 TI - Tumor angiogenesis factors produced by cancer cells. AB - Tumor angiogenic activity from tumor angiogenesis factors (TAFs) produced by 25 cell lines was assayed onto chorioallantoic membranes (CAMs). Neovascularization occurred prominently in such cell lines, as HTBOA (poorly differentiated ovarian carcinoma), HUOCA-II (poorly differentiated clear cell adenocarcinoma), HWUA (poorly differentiated endometrial adenocarcinoma), HKUS (uterine cervical small cell carcinoma), and in HOTHC (anaplastic thyroid carcinoma). The cell lines which secreted TAF showed high heterotransplantability in nude mice and produced rapidly growing tumors which were rich in blood vessels. The TAFs polypeptides of 14,000 and 78,000 molecular weight, were extracted and purified from the conditioned medium of HUOCA-II or W3UF (sub-line of HUOCA-II) lines, respectively. TAFs at concentrations of 10 ng/ml and 100 ng/ml promoted proliferation of the endothelial cells and induced tube formation. Microsequencing analysis revealed that TAF of 78,000 molecular weight has sequence identity with human hepatocyte growth factor (hHGF). PMID- 10457906 TI - [Technology for gene transfer into hematopoietic stem cells (Part 2)]. AB - A hematopoietic stem cell is considered to be one of the ideal targets for gene therapy, and there is expectation that gene therapy will be established based on the technology of hematopoietic stem cell transplantation. However, in recent clinical trials of stem cell gene therapy for monogenic diseases, significant clinical improvement has not been reported. One of the main obstacles is the low efficiency of gene transfer into hematopoietic stem cells. Many investigators have been trying to improve the transduction efficiency to the clinically applicable level. Another approach to solve this problem is to develop the method for selective expansion of transduced hematopoietic stem cells in vivo. We are currently developing novel regulatory genes (selective amplifier genes) for stem cell gene therapy. PMID- 10457907 TI - Species differences in chemical carcinogenesis of the thyroid gland, kidney and urinary bladder. PMID- 10457908 TI - Hormonal imbalances and thyroid cancers in humans. PMID- 10457909 TI - Thyroid stimulating hormone (TSH)-associated follicular hypertrophy and hyperplasia as a mechanism of thyroid carcinogenesis in mice and rats. PMID- 10457910 TI - A mechanistic relationship between thyroid follicular cell tumours and hepatocellular neoplasms in rodents. PMID- 10457911 TI - Human renal-cell carcinoma--epidemiological and mechanistic aspects. PMID- 10457912 TI - Human renal carcinoma--pathogenesis and biology. PMID- 10457913 TI - alpha 2-Urinary globulin-associated nephropathy as a mechanism of renal tubule cell carcinogenesis in male rats. PMID- 10457914 TI - Possible mechanisms of induction of renal tubule cell neoplasms in rats associated with alpha 2u-globulin: role of protein accumulation versus ligand delivery to the kidney. PMID- 10457915 TI - Human bladder cancer: epidemiological, pathological and mechanistic aspects. PMID- 10457916 TI - Calculi, precipitates and microcrystalluria associated with irritation and cell proliferation as a mechanism of urinary bladder carcinogenesis in rats and mice. PMID- 10457918 TI - Agents that induce epithelial neoplasms of the urinary bladder, renal cortex and thyroid follicular lining in experimental animals and humans: summary of data from IARC monographs volumes 1-69. PMID- 10457917 TI - Calcium phosphate-containing urinary precipitate in rat urinary bladder carcinogenesis. PMID- 10457919 TI - Chemicals associated with tumours of the kidney, urinary bladder and thyroid gland in laboratory rodents from 2000 US National Toxicology Program/National Cancer Institute bioassays for carcinogenicity. PMID- 10457920 TI - The role of state-dependent memory in "red-outs". PMID- 10457921 TI - The insanity defense: five years of research (1993-1997). PMID- 10457922 TI - The impact of television on the presentation and reception of children's testimony. PMID- 10457923 TI - Violence and aggression in the heat of passion and in cold blood. The Ecs-TC syndrome. PMID- 10457924 TI - A frustration/negative cue model of unfavorable police treatment of black citizens. Discrimination based on automatic and biased signal processing. PMID- 10457925 TI - Modus operandi and personality disorder in incarcerated spousal killers. PMID- 10457926 TI - Prevalence and severity of lifetime physical and sexual victimization among incarcerated women. PMID- 10457927 TI - Training investigative interviewers: adherence to the spirit, as well as the letter. PMID- 10457928 TI - Psychopathy and offender types. Results from a Portuguese prison sample. PMID- 10457929 TI - Sources of eyewitness identification error. PMID- 10457930 TI - Treatment programs for offenders. Meta-analysis, "what works," and beyond. PMID- 10457931 TI - Public opinion and punitivity. PMID- 10457932 TI - Dangerous offender statutes in the United States and Canada. Implications for risk assessment. PMID- 10457933 TI - [Endoscopic transnasal transsphenoidal approach to the pituitary lesions: anatomy, basic technique and avoidance of pitfalls]. PMID- 10457934 TI - [Brain oxygen extraction fraction as an indicator for shunting operation in normal pressure hydrocephalus]. AB - It is still difficult for neurosurgeons to determine which patients with suspected idiopathic normal pressure hydrocephalus (NPH) should undergo shunting. We need to find a more accurate indicator to predict the effect of shunting. We introduced a new preoperative examination of brain oxygen extraction fraction (OEF) and examined whether preoperative OEF value is effective for prediction of the surgical results. Global brain OEF was calculated from oxygen contents of arterial blood (AO2) and jugular venous blood (VjO2) selectively sampled from the right jugular bulb using the Seldinger method: OEF = (AO2-VjO2)/AO2. Since June 1996 we have treated 9 patients suspected of idiopathic and 10 patients suspected of secondary NPH. OEF in non-NPH patients with dilated ventricle (n = 10) and in infarct patients (n = 85) were 0.33 +/- 0.02 and 0.38 +/- 0.06, respectively. In contrast, OEF increased both in idiopathic NPH (0.42 +/- 0.04) and secondary NPH (0.45 +/- 0.02), and the NPH patients with the higher preoperative OEF values showed the better symptomatic recovery. The present study suggests that brain function may be reversible when OEF can be increased and that OEF can be a useful indicator for predicting the effect of a shunting operation in NPH. PMID- 10457935 TI - [Cerebral blood flow (CBF) before and after cranioplasty performed during the chronic stage after decompressive craniectomy evaluated by xenon-enhanced computerized tomography (Xe-CT) CBF scanning]. AB - Patients who undergo decompressive craniectomy for the purpose of prevention of cerebral herniation sometimes improve neurologically or increase activity of daily life after cranioplasty during the chronic stage. We studied the effect which cranioplasty at the chronic stage had on intracranial environment by means of xenon-enhanced computerized tomography (Xe-CT) CBF scanning. Eight patients underwent decompressive craniectomy at the acute stage and cranioplasty at the chronic stage was reviewed. Xe-CT CBF scanning was performed before and after cranioplasty. On the slice of the basal ganglia. CBF of the symptomatic hemisphere increased in five patients and CBF of the other hemisphere increased in three patients after cranioplasty. On the slice 20 mm above the basal ganglia, CBF of the symptomatic hemisphere increased in five patients and CBF of the other hemisphere increased in four patients after cranioplasty. Cranioplasty after decompressive craniectomy may increase CBF of not only the symptomatic hemisphere but also the other hemisphere. PMID- 10457936 TI - [Simulation study on parent artery occlusion therapy for basilar head giant aneurysms: hemodynamic efficacy of additional bypass on enhancing aneurysmal thrombosis]. AB - Parent artery occlusion therapy has been used as an alternative for directly untreatable basilar head giant aneurysms. However, some aneurysms still require additional technique to further induce intraaneurysmal stagnation and thrombosis. Using a hydraulic model, half-life of the dye in the model aneurysm was measured as an index of the intraaneurysmal stagnation. The ratio of diameters (D1, D2) of the two posterior communicating arteries (PCom) is defined as diameter ratio (DR) (D1/D2, D1 < D2). Occlusion therapy was simulated in 4 different sites. The most distal site was at the distal BA (Type A) and the most proximal site at the bilateral VA (Type D). The additional technique investigated was bypass placement to the P2 segment of the posterior cerebral artery (PCA) on the smaller PCom side. The half-life was 2.5 +/- 0.2 (mean +/- 2SD) sec before occlusion. Occlusion therapy significantly increased the half-life depending on the occlusion site and DR values with the maximum values of infinite in Type A and 25 sec in Type D. Placement of the bypass decreased the tangential flow volume to the aneurysmal neck, and increased the time markedly in Types A and B, considerably in Type C and slightly in Type D. Parent artery occlusion therapy has been considered less beneficial for patients with a diameter ratio of less than 0.70 since aneurysmal stagnation depends on the diameter ratio. Bypass placement additional to the occlusion therapy is very useful in enhancing the aneurysmal stagnation, which would make those patients indicated for the therapy. PMID- 10457938 TI - [The usefulness of dynamic computed tomography scanning in predicting the outcome of acute stroke patients]. AB - The aim of this study was to investigate whether brain dynamic computed tomography (CT) is useful in predicting clinical outcome. Thirty patients suffering from cerebral ischemia in the territory of the middle cerebral artery (MCA) underwent dynamic CT scanning within 6 hours of stroke onset. Regions of interest (ROIs) were placed in the bilateral MCA territories and three parameters, peak value (PV), time to peak (TP), and PV divided by TP, were calculated from time-density curves (TDCs) on ROIs. After conventional treatment using pharmacological agents, the 30-day clinical outcome was evaluated on the Glasgow outcome scale. To investigate the relationship between the disease-to contralateral side ratio of each parameter's value and 30-day clinical outcome, TDCs were classified into the following four types; type 1, with TP ratio less than 1.1; type 2, with TP ratio ranging from 1.1 to 1.5 and PV/TP ratio more than 0.75; type 3, with TP ratio ranging from 1.1 to 1.5 and PV/TP ratio less than 0.75; and type 4, with TP ratio more than 1.5 and PV/TP ratio less than 0.3. Clinical outcome in patients with type 1 or 2 TDC was better than in patients with type 3 or 4 TDC (p < 0.01, Fisher's exact test). We can conclude that dynamic CT is a useful means for estimating the clinical prognosis of acute stroke patients after conventional treatment. Poor clinical outcome following conventional therapy is expected in patients with type 3 or 4 TDC in contrast to patients with type 1 or 2 TDC. PMID- 10457937 TI - [Prevention of MRSA spread in the neurological field: intranasal application of mupirocin calcium ointment]. AB - From September 1997 to March 1998, forty patients with cerebral disorders were investigated. They were divided into two groups: one treated and the other untreated. Mupirocin calcium ointment (MCO) was applied three times a day for three days into the nasal cavities of the patients in the treated group. In order to check the growth of MRSA (methicillin-resistant Staphylococcus aureus), bacterial isolation culture from the nasal cavity was carried out on admission, one week after admission and one month after admission. MRSA was nor detected in isolation culture of any of the cases on admission. One week later MRSA was detected in isolation culture of one case of the 20 MCO treated patients and in three of the 20 untreated patients. There was no significant difference between treated and untreated groups. In isolation culture after one month, MRSA was recognized in four cases of 16 in the MCO treated group (three patients were discharged and one expired). On the other hand, it was recognized in eight cases of thirteen in the untreated group (seven cases were discharged). MRSA infection of the nasal cavity decreased significantly due to MCO treatment (p < 0.05). It is suggested that the nasal carriage of MRSA was prevented by intranasal application of MCO on admission. PMID- 10457939 TI - [Diagnosis and treatment of nontraumatic dissecting aneurysm in the middle cerebral artery]. AB - Two cases of nontraumatic dissecting aneurysm of the middle cerebral artery (MCA) are reported. A 59-year-old woman presented with subarachnoid hemorrhage, mainly in the right sylvian fissure. On admission, the right carotid angiogram revealed a dissecting aneurysm with a double lumen extending from segment M1 to M2 of the right MCA, and an unruptured saccular aneurysm in the right internal carotid artery. Emergency surgery revealed a discolored protrusion of the arterial wall in the right MCA, which was thought to be the cause of her subarachnoid hemorrhage. The protrusion of the arterial wall was clipped and coated with Bemsheet soaked in Biobond. However, disturbance of consciousness persisted and she died of paralytic ileus two months after the operation. The other patient was a hypertensive 33-year-old woman with right hemiparesis and motor dysphasia. CT scans obtained on the day of admission showed no abnormalities. She was treated conservatively with clinical improvement, but CT scans obtained 3 days after the ictus revealed an infarction deep in the left frontal lobe. A left carotid angiogram was made 4 days after ictus and demonstrated severe stenosis of the proximal segment of the left MCA with poor filling of its superior trunk. Despite improvement of her hemiparesis, CT scans obtained 3 weeks after the ictus showed hemorrhagic infarction in the left frontal lobe. Repeat left carotid angiogram revealed a double lumen in the C1 and M1 portions with improvement of the previous severe stenosis of the M1. The 23 reported cases of DA in the MCA with our cases are reviewed and their neuroradiological and clinical features are discussed. PMID- 10457940 TI - [Intracerebral hemorrhage in a patient with moyamoya phenomenon caused by tuberculous arteritis: a case report]. AB - A case of intracerebral hemorrhage which probably had been caused by a rupture of abnormal "moyamoya vessels" due to tuberculous arteritis was reported. A 42-year old female had a history of tuberculous meningitis at the age of 2 years and suffered from a sudden onset of severe headache in January of 1998. CT scan disclosed a medium-sized intracerebral hematoma in the left frontal base and many calcifications in the basal cistern. Subsequent angiography demonstrated high grade stenosis in the terminal portion of the right internal carotid artery and near-by "moyamoya vessels". No surgery was performed on the patient. A second angiography was carried out two months later and it newly disclosed ophthalmic artery-feeding "moyamoya vessels" in the place where the intracerebral hematoma had been located. This led us to conclude that a rupture of "moyamoya vessels" was the cause of the intracerebral hemorrhage and "moyamoya vessels" were not visible in the first angiograms because they had been compressed by the hematoma. Although cerebral infarction is common in tuberculous arteritis, cerebral hemorrhage is uncommon. The pathogenesis of cerebral hemorrhage due to tuberculous arteritis and its difference from that of hemorrhage caused by moyamoya disease is discussed. PMID- 10457941 TI - [Ruptured middle cerebral artery aneurysm successfully treated by intra aneurysmal GDC embolization in a patient with idiopathic thrombocytopenic purpura: a case report]. AB - We report a case of a ruptured middle cerebral artery (MCA) aneurysm treated by an intra-aneurysmal Guglielmi detachable coil (GDC) embolization in a patient with idiopathic thrombocytopenic purpura (ITP). A 57-year-old female, who had been suffering from ITP for 4 years, experienced sudden severe headache. She was diagnosed as having subarachnoid hemorrhage due to a ruptured MCA aneurysm and the hematological examination showed moderate thrombocytopenia. Considering the risk of open surgery, we treated the aneurysm by intra-aneurysmal GDC embolization in the acute stage and platelet transfusion was carried out after the therapy. She had a good clinical course and recovered without any complications or neurological symptoms. The follow-up cerebral angiogram at 1 week and 2 years after the treatment showed complete obliteration of the aneurysm. Recently, endovascular treatment tends to be applied to surgically high risk patients. A GDC embolization for a patient with thrombocytopenia has rarely been reported and the risk and efficacy is unknown. So, we conclude that for a patient with thrombocytopenia, intra-aneurysmal GDC embolization could be adopted as one of the less invasive treatment for ruptured aneurysm. PMID- 10457942 TI - [A case of frequent intracerebral hemorrhage from mixed vascular malformation in a child]. AB - We reported a rare case of frequent intracerebral hemorrhage from mixed vascular malformation in an infant. A 1-year and 4-month-old girl was admitted with seizure. She had been diagnosed as having febrile convulsion at the age of 8 months. Computed tomography (CT) indicated a multicystic mass without perifocal edema in the right frontal lobe. The mass was not enhanced by contrast media, but T1-weighted magnetic resonance imaging (MRI) revealed a multicystic mass with mixed signal intensity, while T2-weighted image demonstrated a heterogenous high signal intensity mass with a marginal low intensity rim. These findings suggested a cavernous angioma causing intracerebral hematomas. Because of this, total resection was achieved via a right fronto-temporal craniotomy without neurological deficits. Histologically, numerous dilated venous-type blood vessels interspersed within the normal brain parenchyma were seen. In parts, the vessel walls were abnormally thickened and the lesion coalesced with capillary-like vessels. We diagnosed mixed vascular malformation, mainly composed of venous malformation along with telangiectasia. The number of reports for this so-called mixed vascular malformation have thus been increased. The histopathological criteria for classification of vascular malformation should be reinvestigated carefully. PMID- 10457943 TI - [Normal aging and abnormal aging of the brain]. PMID- 10457944 TI - [Functional aging of the central nervous system]. PMID- 10457945 TI - [Physiological atrophy and pathological atrophy: morphological aspect in brain aging]. PMID- 10457946 TI - [Biochemical aspects of aging]. PMID- 10457948 TI - [Progress of neuroendoscopy: current application and future perspectives]. PMID- 10457947 TI - [Mitochondria and aging]. PMID- 10457949 TI - [Clinical study on intracranial air in the severe head injury]. AB - Intracranial air tends to be neglected in severe head injury, because other lesions are more life threatening. We retrospectively studied patients of severe head injury with pneumocephalus. We studied 250 patients' (Glasgow Coma Scale score < or = 8 on admission) computed tomography (CT) scans and found 77 patients with pneumocephalus. These patients' medical records and neuroimagings were studied. The air was most frequently located in the subarachnoid spaces followed by subdural and epidural spaces. Intraparenchymal and intraventricular location were rare. Sphenoid or petrous bone fracture was common. In the cases with petrous or paranasal sinus bone fracture, opacification of these structures were statistically significant. In conclusion, pneumocephalus is common in the severe head injury and most frequently located in the subarachnoid spaces, however, the precise location of air is unclear in many cases. Bone CT scan is useful and opacification of air containing structures suggests the possibility of skull base fractures. PMID- 10457950 TI - [Evaluation of cranial CT findings of patients with muscular dystrophy: with a reference to cerebral vascular disease and cardiac complications]. AB - We evaluated cranial CT findings of 160 patients with various type of progressive muscular dystrophy (PMD). Significant brain atrophy was observed in 21 out of 63 cases of Duchenne muscular dystrophy (DMD), 7 out of 15 Becker muscular dystrophy (BMD), no case of 2 female dystrophinopathy (F-dyst), 11 out of 21 limb-girdle muscular dystrophy (LG), all cases of 10 Fukuyama type congenital muscular dystrophy (FCMD), 2 out of 5 fascioscapulohumeral muscular dystrophy (FSH), and 32 out of 44 myotonic dystrophy (MyD). Genetical degenerative process and vascular insufficiency seemed to cause brain atrophy in these disease. The intracranial calcification was observed in one DMD, one LG and seven MyD. One LG patient showed focal atrophy in left temporal lobe, and one MyD demonstrated right temporal meningioma. The trace of cerebral vascular accident was disclosed in eleven patients with PMD (1 DMD, 2 BMD, 1 F-dyst, 2 LG, 5 MyD). In these cases, 2 patients had dilated cardiomyopathy, 6 patients with decreased left ventricular ejection fraction, 3 with atrial fibrillation, 1 with cardiac arrest followed by pacemaker instillation, 1 with Adam-Stokes attack, and 3 with 1 degree AV-block. Diffuse low density in the white matter was seen in a patient with F-dyst, a FCMD patient, and 8 MyD patients. Cardiac emobolism, severe arrythmia, cardiogenic shock and hemodynamic disorder were seemed to cause cerebral vascular disease in PMD. PMID- 10457951 TI - [Germinal matrix hemorrhage and hydrocephalus in premature infants]. AB - An analysis of 25 preterm infants with grade 3 or 4 intraventricular hemorrhage and surgically treated progressive hydrocephalus was undertaken to assess mortality, cognitive outcome and motor deficit. All patients underwent surgery at the author's institute since 1981 and represented 4.7% of all intraventricular hemorrhage in premature infants. Surgical procedure was initial placement of a miniature Ommaya's reservoir and conversion to a ventriculoperitoneal shunt diversion. Overall, outcomes were achieved in 64.7% of ambulatory and improvement of cerebroventricular index on follow-up CT image from 0.68 +/- 0.36% to 0.36 +/- 0.31%. The presence of intraventricular hemorrhage continues to be a major problem in low birth weight infants whose rate survival continues to increase as the major improvements in perinatal medicine. We feel that miniature Ommaya's reservoir is quite helpful in the treatment of posthemorrhagic hydrocephalus in the preterm infant who prove refractory to aggressive conservative therapy. PMID- 10457952 TI - [A case of atypical type of Sturge-Weber syndrome demonstrated reversible change by MRI FLAIR method in ictus and in postictal state]. AB - We report a patient of atypical type of Sturge-Weber syndrome who demonstrated a reversible change by MRI FLAIR method in ictus and postictal state. A 5-year-old boy was admitted to our hospital because of severe headache, vomiting and loss of consciousness with his eyes conjugated to left for a few minutes. He had no facial nevus and other abnormal findings in physical examination. CT scan showed two small calcifications in the right occipital lobe. Postcontrast T 1-weighted image of MRI demonstrated a right parieto-occipital leptomeningeal enhancement. We diagnosed this case as an atypical type of Sturge-Weber syndrome. Although, on admission, FLAIR method showed the area of high signal intensity, after anticonvulsant therapy, those abnormal area disappeared. It is presumably detected by FLAIR method slight extravasation of plasma element in the surface of the brain due to regional hyperperfusion in ictus. PMID- 10457953 TI - [Vitamin B1 deficiency polyneuropathy presenting homolateral imitative synkinesia]. AB - We report a 56-year-old woman with vitamin B1 polyneuropathy, showing bilateral homolateral imitative synkinesia. Needle electromyogram revealed neurogenic changes, and the amplitude of muscle action potential was low. Sural nerve biopsy showed a marked loss of myelinated fibers, and severe axonal degeneration was diagnosed. Spinal and brain MRI revealed no abnormalities. In the literature, these synkinesias were observed in patients with parietal, thalamic, and basal ganglia lesions and with chorea. We suggest that this synkinesia is the release phenomenon in the circuit of the motor programming system due to the disturbance of peripheral nerve or funiculus posterior. PMID- 10457955 TI - [Hemangioma of the frontal bone]. PMID- 10457954 TI - [Two cases of fronto-temporal dementia without remarkable lobar atrophy]. AB - Front-temporal dementia (FTD), advocated by Lund and Manchester groups, includes Pick type corresponding to the conventional frontal Pick's disease, motor-neuron type associated with neural symptoms, and frontal lobe degeneration type. In Japan, however, there have been few case reports of the frontal lobe degeneration type. Here we examined clinical characteristics and imaging findings of 2 cases of FTD frontal lobe degeneration type. Neurological examinations were normal. CT and MRI scans revealed no obvious frontal lobar atrophy, while HMPAO-SPECT scans demonstrated remarkable hypoperfusion in anterior hemisphere. Neuropsychological examination revealed frontal symptoms, including personality change, stereotypes, and disinhibition. These symptoms can not be distinguished from the cases of FTD Pick type, because those cases of FTD Pick type have obvious lobar atrophy. PMID- 10457956 TI - [MRI of the impending cerebral herniation caused by chronic subdural hematoma]. PMID- 10457957 TI - Cerebral vascular accident after cardiac surgery: its impact on nursing care. AB - The number of patients undergoing coronary artery bypass grafting (CABG) per year is increasing. Despite advances in surgical techniques, cerebral vascular accident (CVA) post cardiac surgery is increasing. CVA is a severe neurological complication of cardiac surgery which increased length of stay, morbidity and mortality, and rehabilitation. It is important to identify patients at increased risk and utilize appropriate screening techniques to decrease the incidence of CVA. Nursing assessment, interventions, and postoperative neurological assessment is crucial in identifying patients at increased risk for CVA. PMID- 10457958 TI - How men and women with heart disease seek care: the delay experience. AB - Coronary heart disease (CHD) is the leading cause of death among American men and women. Much research has focused on CHD in men while less is known about the disease in women. Traditionally, research protocols have excluded women or have not addressed gender differences in the experience of the disease. A major factor contributing to morbidity and mortality is delay in seeking medical care when experiencing symptoms of CHD. The purpose of this study was to examine the experiences of men and women with symptoms of CHD in seeking care. A sample of 121 men and women admitted to one of two cardiac units in an urban hospital were interviewed using a questionnaire concerning demographics and factors related to delay. The study demonstrated that women were more likely than men to contact a family member when experiencing symptoms. Smokers and previous smokers with symptoms were less likely to delay seeking care. The findings suggest important differences in the experience of CHD among men and women. Implications for practice and directions for further research are discussed. The information gained can be used to develop interventions that reduce delay and improve health care for men and women with CHD. PMID- 10457959 TI - Cardiovascular effects of acute stroke. AB - A wide variety of dysrhythmias, conduction defects, and repolarization abnormalities have been observed following all types of stroke, most likely related to disruptions in the autonomic nervous system. Patients at highest risk for developing ECG changes include those with hemorrhagic stroke, those with strokes involving the right cerebral hemisphere, and elderly patients with stroke. Nursing interventions for these patients revolve around vigilant monitoring of both neurologic and cardiovascular status, to identify any negative consequences from the interaction of these systems. Strict blood pressure monitoring is required, with different management goals depending upon the type of stroke. Also important is the prevention of further exacerbation of ECG abnormalities by maintaining normal electrolyte levels. PMID- 10457960 TI - The thread that links: continuity of patient care. AB - With managed care shortening hospital stays, nurses at the bedside are challenged to come up with innovative ways to utilize time wisely. Our critical care transport team has implemented a strategy that assists in this endeavor. An universal admission data sheet is completed when each patient arrives on the unit. This form takes about 45 minutes to 1 hour to complete. It identifies several issues that impact on patient teaching and discharge planning. By completing this form enroute, we provide the bedside nurse with more time to concentrate on discharge planning and teaching. Follow up indicates that early implementation of this form leads to a shorter, more productive stay for the patient. We have also observed an improvement in the continuity of patient care, since the transport nurse completing the form communicates the information to the primary nurse receiving the patient. This has also led to a strong link between the outside satellite staff and the base hospital staff through better relationships between the transporting and receiving nurses. PMID- 10457961 TI - Drug absorption in congestive heart failure: impact on management. PMID- 10457962 TI - What is the most serious complication for patients with anorexia nervosa? PMID- 10457963 TI - Apo (E) gene: another independent risk factor for coronary artery disease? PMID- 10457964 TI - [Societal perception of and experience with psychopharmaceutical drugs]. PMID- 10457965 TI - [Scientific psychology in the works of Eugen Bleuler]. AB - The name of Eugen Bleuler is usually known in the context of the definition and conceptualization of schizophrenic disorders. Main other topics, less known in literature, are alcoholism, forensic psychiatry and general psychological topics of meaning of consclousness, formation of motive and will. Bleulers point of view presents itself as an empirically-based and anti-philosophic one, definitely deterministic concerning the topic of human "freedom of will". Eugen Bleuler refers to an already (especially by Richard Semon) systematically developed theory called "mnemism" that he interpreted and applied to the psychological circumstances mentioned above. That theory of "mnemism", that can be most adequately described as a biogenetic-vitalistic theory, is assuming, that all organic life--independent of the possibility of a self-reflecting consciousness- is able to learn experiences made by analysis of environment and to pass it on following generations. Pattern of stimulus reactions are in the sense of this theory memorized and reactivated under similar situational circumstances by the psychological mode of association. The influence of psychoanalytical theories, especially the presumed importance of infantil sexuality, on Bleulers understanding of a medical psychology seems to fade over the years. PMID- 10457966 TI - [Diagnosis and therapy of depressive disorders in general practice]. AB - 93 general practitioners participating in a WHO field trial testing the ICD-10 primary health care classification system in German-speaking countries completed a questionnaire dealing with their practice of diagnosis and treatment of patients with depressive disorders. There was a great variety of diagnoses that were often vague. Counselling tended to be "good advice" instead of specific intervention. Antidepressant medication mostly consisted of tricyclic antidepressants in rather low dosages. Only few of the general practitioners informed patients thoroughly about side-effects and risks of antidepressant medication. PMID- 10457967 TI - [DEWIPA--a standardized questionnaire for assessing knowledge about symptoms, etiology and psychopharmacologic treatment in patients with depressive episodes]. AB - OBJECTIVE: The aim of this study was to develop a questionnaire for measuring depressed patients' knowledge of their disease. METHODS: 55 patients suffering from depressive episodes answered a 77-items questionnaire. RESULTS: Item analysis showed a discriminative power higher than 30 for 40 items, all of which applied to one factor. The items were arranged into two thematically similar 20 items forms. We added four additional items (discriminative power between 0.20 and 0.29) because future research may lead to different results concerning discriminative power. The questionnaires DEWIPA-A and DEWIPA-B consists of 24 items each. The average difficulty is equal for both DEWIPA forms and they both possess significant discriminative power. CONCLUSIONS: Our results support the hypothesis that DEWIPA is a valid and reliable instrument to determine patients' knowledge of symptoms, causes and the psychopharmacological treatment of depression. PMID- 10457968 TI - [Attitude of family to neuroleptics]. AB - PURPOSE: The present study is based on the assumption that for patients' compliance with psychotropic medication is also of importance their closest relatives' attitude towards that treatment. METHODS: In the context of a mail survey of relatives of people with schizophrenia, we asked what the relatives considered to be the most important positive and negative effects of the medication. RESULTS: As expected, the main benefit was seen in the reduction of symptoms and in the prevention of relapse. In addition, the improvement of social adaptation and the subjective quality of life of the patients were also cited as important positive effects. Beside these effects which are closely related to the treatment of the disorder, respondents mentioned effects which can help to facilitate their living together with their ill relative. Among the side effects regarded as particularly unpleasant for the patients, sedation ranked first, followed by weight gain and extra-pyramidal motor symptoms. DISCUSSION: The results are contrasted with those of a comparable study of people with schizophrenia. PMID- 10457969 TI - [Neuroleptics in the newspaper. A mass media analysis]. AB - OBJECTIVE: Neuroleptic drugs have a dubious public reputation. Many people consider them obnoxious, even in schizophrenia. METHOD: 10 annual editions of six German language opinion leading newspapers are analysed. RESULTS: Only in one of 35 articles neuroleptics are judged positive, in 27 negative. They are seen close to violence and restriction, as punishment or even as torture, as paralyzing or even as "torturetherapy". Informations in the articles are scarce. Most of the articles cover the fields of medicine or psychiatry or of forensics. CONCLUSION: Apart from the negative attitude the extremely low number of articles on neuroleptics is the most outstanding result of our study. PMID- 10457970 TI - [Attitude of family to clozapine]. AB - AIM: Evaluations of clozapine treatment by relatives of schizophrenic patients were assessed as part of an exploratory study and compared with the patients' own assessment. METHOD: 46 relatives of individuals suffering from schizophrenia were questioned by means of a problem-centred interview. RESULTS: In addition to the positive effects on the schizophrenic illness, relatives strikingly often pointed out the calming, sleep-improving effect of clozapine. As opposed to the patients, the majority of relatives were well aware of the risks associated with discontinuing the drug treatment. Both patients and relatives complained about the sedative effect and the increased need for sleep associated with clozapine treatment. However, diverging evaluations were given with regard to hypersalivation, anticholinergic effects, and weight gain. Relatives were more concerned about possible organ damage occurring as a long-term effect than the patients. Further, indications were found that relatives were more worried by the risk of haematotoxic effects resulting from clozapine treatment than the patients. It could be said that, among the relatives, there was a slightly greater awareness of the advantages of clozapine treatment in comparison with conventional neuroleptics than there was with the patients. CONCLUSION: The importance of relatives' views in the effort to secure adequate neuroleptic treatment despite the current economic restrictions should not be under estimated. PMID- 10457971 TI - ["Rather overweight and mentally present...". Patients evaluate clozapine]. AB - OBJECTIVE: Subjective evaluations by schizophrenic patients of clozapine treatment were assessed as part of an exploratory study. METHODS: A problem centred interview was carried out with 80 patients at discharge from in-patient or day hospital treatment. RESULTS: In addition to expected effects (improvement or stabilisation of one's state of mental health, antipsychotic effects), patients surprisingly often highlighted clozapine's calming and relaxing effect as well as improved sleep as particularly positive. While more than half of the respondents expected a worsening of their condition in case they stopped taking their medication, only every fifth patient feared a relapse. Among the negative effects, fatigue and sedation were cited by far the most often. The absence of extrapyramidal side effects was clearly noted as an advantage of clozapine. Only every tenth of those questioned was aware of the risks for the haemotopoietic system associated with the drug. With regard to the obligatory blood tests, a surprisingly large number of patients was indifferent or stated that they had no opinion on the matter. DISCUSSION: Possible implications for patient information and compliance will be discussed. PMID- 10457972 TI - [Tranquilizers in the newspaper. An media analysis]. AB - OBJECTIVE: For the public the image of psychopharmacological drugs is identical with that of minor tranquilizers. The way of describing and portraying these drugs in the media therefore is of general interest. METHOD: Ten annual volumes of six German language opinion leading daily and weekly papers which exist on CD Rom in full text editions are screened for the key word "minor tranquilizer". The articles identified are analyzed formally quantatively and qualitatively. RESULTS: The ten annual volumes analyzed contained 45 articles in which tranquilizers were mentioned (4.5 per annual volume). The key words were found in all sections of the papers, most often, however, in the cultural section, followed by the science section. Regarding the themes medicine and law were in the first position (one third), but society, arts and culture and politics contained even more articles dealing with the key word minor tranquilizer. For fifth of the articles only mentioned the key word. Only once minor tranquilizers are the main topic of the article. Tranquilizers are valued negatively ten times as often as they are valued positively. In two fifth of the articles the term is employed as a metaphor. Even in this context minor tranquilizers are valued negatively (10:1). CONCLUSIONS: Three fourth of the articles, employing the term in the non metaphorical way, deal with abuse and dependency. The public image of minor tranquilizers in the media is mainly characterized by their potency to cause dependency. PMID- 10457973 TI - [Biological addictive drugs--is there a change in drug taking behavior of young drug addicts?]. AB - Abuse of natural products was frequently noticed among young patients with multiple substance abuse. 180 patients of a rehabilitation clinic for young addicts filled out a questionnaire regarding their regularly consumed drugs. Ayahuasca and cactus, the natural drugs of the 70's are rarely found. The patients frequently consumed psilocybin, amanita and datura. We inform about the toxicological basis of the used drugs. A specific exploration of patients with multiple substance abuse in regard to the use of biological drugs is an necessary as random and multicentre follow-up-studies. PMID- 10457974 TI - [Acute life-threatening catatonia]. AB - Malignant catatonia is a rare, life-threatening syndrome that occurs in connection with somatic as well as psychiatric disorders, in particular with functional psychosis. Key symptoms are catatonic features combined with elevated muscle tone, hyperthermia, vegetative instability, and pathological laboratory values. Benzodiazepines, dantrolene, and ECT have been reported to be an effective treatment of malignant catatonia, whereas classic neuroleptics and anticholinergic drugs are contraindicated. If antipsychotic treatment is required, clozapine is generally used. In this article we describe a case of malignant catatonia in a 52 year old female patient and discuss the clinical picture, the treatment, and the time course of the syndrome. PMID- 10457975 TI - [Symptomatic psychosis in bland sinus thrombosis and antibiotic therapy]. AB - It is difficult to define intracranial sinus thrombosis by clinical symptoms only. We report on a case of symptomatic psychosis which occurred in sinus thrombosis with defective blood-brain barrier during treatment with antibiotics. PMID- 10457976 TI - [Dealing creatively with laws regarding protective custody]. PMID- 10457977 TI - [Therapeutic problems of positively experienced hallucinations]. PMID- 10457978 TI - A comparison of social functioning among black and white women with breast cancer. AB - Results of a comparative study of interviews with 102 women (61 white, 41 black) who were treated for breast cancer suggest that black women have more difficulty in social functioning, especially the resumption of household activities. Implications for social work practice are discussed. PMID- 10457979 TI - Coping with pediatric migraine. AB - Strategies used by latency-aged children to cope with pediatric migraine pain are identified in this exploratory study. The following three broad categories of coping and their subtypes emerged from the data: affective, cognitive, and problem-focused coping. Implications for practice, research, and education, particularly regarding the use of spirituality and prevention, are discussed. PMID- 10457980 TI - Advance directives: the experience of health care professionals across the continuum of care. AB - This study explored the perceptions of health care providers across the continuum of care as to how effective advance directive arrangements were in assuring compliance with the patients' wishes, as well as their overall satisfaction levels with the process. The health care providers who responded to the survey indicated high levels of overall satisfaction with advance directives, despite low patient completion rates and, most significantly, low confidence levels that surrogate decision making accurately reflected the patient's wishes. A secondary analysis compared the perceptions of health care providers with consumers' reasons for failing to complete advance directives. The need for further education was perceived as a primary need by the health care professionals but not by the consumers. Recommendations for policy initiatives and further research are then presented. PMID- 10457981 TI - Organizing social work services with adult cancer patients: integrating empirical research. AB - Today only half of those diagnosed with cancer will die of the disease, leaving enormous room for psychosocial interventions to improve the psychological or functional status of those coping with the disease. Therefore, social workers in oncology must be current with empirical research. In an effort to integrate current research into social work practice, we reviewed empirical studies with sound research designs to answer the following questions: (1) What portion of cancer patients are likely to need social work services? (2) What types of services do social workers provide to meet these needs? and (3) Who is likely to be (or not to be) the recipient of these services. Does intervention research include diverse (non-traditional, non-white and non-middle class) clients? The results show about one third of patients will be judged at high-risk for psychosocial problems but that only 15-25% of those who are diagnosed with cancer will eventually use psychosocial oncology services. Some have mainly instrumental, concrete needs, and others will use psychosocial counseling. A review of the inclusion of minorities and non-traditional, non-middle-class groups, shows that they are not adequately represented in current intervention research in psychosocial oncology. Methods for enlarging their access and participation are suggested. PMID- 10457982 TI - Measuring quality of life in back patients: comparison of Health Status Questionnaire 2.0 and Quality of Life Inventory. AB - This paper is a reliability and validity test of the Health Status Questionnaire (HSQ) 2.0. In addition, the Quality of Life Inventory (QOLI) is compared with the HSQ 2.0 to assess concurrent validity. The study is unique because these instruments are assessed for the first time using a sample of chronic back patients. Practitioners will therefore now be able to evaluate important quality of life issues and treatment changes in this population. The HSQ 2.0 was generally found to be valid for these patients. However, two scale domains were not differentiated due to unique characteristics of spinal disease patients. The QOLI, due to the limitation of measuring only psychological well-being, did not measure problems specific to back patients. PMID- 10457983 TI - Hormone replacement therapy: women's decision-making process. AB - Making an informed decision about whether to take hormone replacement therapy to offset short- and/or long-range effects of decreased estrogen due to menopause is particularly important because of the health-related risks and benefits associated with hormone replacements. A qualitative study was conducted in which a cohort of menopausal women were interviewed for the purpose of investigating their decision-making process regarding HRT. Findings illuminate the major factors contributing to the decision, the kinds and sources of information used in arriving at a decision, the decision-making process followed, and whether the decision was informed. Clinical and research implications of the findings are examined. PMID- 10457984 TI - [Natural and synthetic inhibitors of thrombin. I. Natural inhibitors]. AB - Structures and properties of main physiological (heparin, antithrombin III, heparin cofactor II) and nonphysiological (hirudin, thrombin-binding aptamers, cyclotheonamides) natural thrombin inhibitors and its fragments and synthetic analogs (hirugen, hirulogs, hirunorms, pentasaccharides, macrocyclic alpha-keto amides) were reviewed. The molecular mechanisms of interaction of these compounds with thrombin and their anticoagulant activity at preclinical and clinical trials are discussed. The examined of natural thrombin inhibitors and their synthetic fragments and analogs are perspective for prophylaxis and treatment of different thrombo-embolic diseases. PMID- 10457985 TI - [Natural and synthetic inhibitors of thrombin. II. Synthetic low-molecular-weight inhibitors]. AB - The up-to-date problems, concerning the structure and properties of two types of inhibitors are reviewed. It is particularly considered properties of low molecular weight thrombin inhibitors that have electrophilic groups capable to react with Ser-195 of thrombin (peptidyl-chloromethyl ketones, aldehydes, ketomethylene derivatives and derivatives of boric and phosphoric acids) and the competitive reversible thrombin inhibitors. The review focuses on methods of modification of the structure in the natural inhibitors and design of new peptidomimetics. The prospects for prophylaxis and treatment of diverse thromboembolic diseases are discussed. PMID- 10457986 TI - [The C(27)-steroid hormones ecdysterone and calcitriol activate the phosphoinositol cycle in its membrane phase]. AB - We have examined in vivo the capacities of two C27-steroid hormones phytoecdysteroid ecdysterone (20-hydroxyecdysone, 10(-8) M) and calcitriol (1 alpha, 25-dihydroxyvitamin D3, 10(-12) M), as a modulators of phosphoinositide cycle in the brain and heart cells. Severe lines of evidence indicate that both hormones may acts as positive regulators of phosphoinositide hydrolysis by phospholipases C and D in membrane (0.5-30 min) pre-genomic phase of its actions. Thus, our findings identify ecdysterone, like calcitriol, as a positive regulator of [Ca2+]i, and protein kinase in the mammalian cells. PMID- 10457987 TI - [Inhibitory effect of linoleyl-hydroxamic acid on the oxidation of linoleic acid by 12-lipoxygenase from porcine leukocytes]. AB - Linoleic acid oxidation by 12-lipoxygenase from porcine leukocytes has been studied as affected by linoleyl-hydroxamic acid. Linoleyl-hydroxamic acid has been found to be an effective inhibitor of porcine leucocyte 12-lipoxygenase. Aerobic preincubation of 12-lipoxygenase with 0.1-6 microM of linoleyl-hydroxamic acid led to a time- and dose-dependent inhibition of the enzyme. The inhibitor's concentration able to induce a 50% loss of the enzyme activity with and without 15-min preincubation were 3.5 and 0.65 microM, respectively. Experimental results obeyed a kinetic scheme, which supposed 2 extra substrate molecules bounding with the enzyme-substrate complex in the presence of linoleyl-hydroxamic acid. PMID- 10457988 TI - [Role of phospholipids in the regulation of activity of porcine leukocyte 12 lipoxygenase]. AB - 12-Lipoxygenase from porcine leukocytes was partially purified by using of DEAE Toyopearl chromatography (pH 7.5). Phosphatidylcholine and Phosphatidylinositol in reaction mixtures with mixed micelles Lubrol PX/linoleic acid inhibited the enzyme. The pH-optimum of lipoxygenase reaction in presence of phospholipids shifted into alkaline region. In the absence of phospholipids 3 additional substrate molecules bound with enzyme-substrate complex. In the presence of either phosphatidylcholine of phosphatidylinositol up to 2 substrate molecules bound with enzyme-substrate complex. The phospholipids competed with linoleic acid for one of the enzyme binding centers. A kinetic scheme of 12-lipoxygenase reaction has been proposed: Phosphatidylinositol lowered the values of Ks and Kns of the reaction of linoleic acid oxidation by 12-lipoxygenase, while phosphatidylcholine had opposite effect on these parameters. We suppose that phospholipids can regulate 12-lipoxygenase activity via control of the enzyme affinity to the substrate (polyunsaturated fatty acid). PMID- 10457989 TI - Effect of RoseOx on peroxidation of rat mitochondrial lipids. AB - The effect of the RoseOx drug on rat liver mitochondrial lipids free-radical oxidation in vivo was studied. During the period of 14 days 50 mG/kg per body weight of RoseOx was added to the diet of normal rats each day. Free radical oxidation in liver mitochondrial fraction was determined by the help of a chemiluminescence method. Four kinetic The RoseOx addition to the usual diet led for free radical oxidation braking in mitochondrial fraction of liver as was shown. The RoseOx antioxidizing effect was stipulated by availability of carnosic acid as a supplement. One of the mechanism of the caronosic acid antioxidizing action could be its participation in LFRO reactions breaking by its OH-groups. Carnosic acid contains OH-groups in its molecule as well as a vitamin E for example. So, the mechanism of carnosic acid antioxidizing action is probably similar to vitamin E action in lipid free radical oxidation reactions. PMID- 10457990 TI - [Membrane lipids of brush border of the rat small intestine as affected by ionizing radiation]. AB - It was established that single total X-ray irradiation in the doses of 0.1; 0.4; 1.0; 2.0; 3.0 and 6.0 Gy 24 hours after irradiation results in reliable changes in membrane lipids composition of brush border of enterocytes in doses over 1.0 Gy. By this changes under increase of dosage of irradiation it were marked differences in comparison with control in lipid-protein, total phospholipids protein, cholesterol-protein and cholesterol-total phospholipids rations. In lipid composition major changes are connected with increase of lysophosphatidylcholine and lysophosphatidylethanolamine concentrations, decrease of sphingomyelin content and increase of that of phosphatidylethanolamine. Content of cholesterol and free fatty acids decreased reliably under irradiation in doses over 1.0 Gy as well. Data obtained proves that structural-functional properties of brush border membranes of enterocytes of small intestine are altered under irradiation in doses ranging from 1.0 to 6.0 Gy. Lower doses (0.1; 0.4) cause only trend of changes named above. PMID- 10457991 TI - [Effect of incorporated ionizing radiation by cesium-137 on the state of free radical processes and the lipoprotein spectrum in rat blood]. AB - In vivo experiments it was shown that the prolonged incorporated low levels radiation by 137Cs leads to changes in free-radical processes in blood and in lipoprotein exchange. It is considered that such changes could be very important for postradiation atherogenesis, as possible distant nonstochastic consequences of Chernobyl accident. PMID- 10457992 TI - [Effect of Cu2+ and Ni2+ on the intensity of RNA synthesis in corn seedlings]. AB - It has been found that Cu and Ni at contents of 17 mg/kg and 64 mg/kg dry weight respectively in leaves tissues have no visible effect on morphological and physiological characteristics of plants and increase [3H]uridine incorporation in various types of RNA. Under these conditions synthesise of nuclear RNA has been found 1.8-2.4 more intense, transport RNA -2.0-2.8 intense and ribosomal RNA-1.2 2.8 intense in comparison with control. Analysing our data it is possible to suggest that the activation of the RNA synthesis under the heavy metals influence may be one of the protective mechanism connected with the plant cell adaptation to the stressful conditions. PMID- 10457993 TI - [Proliferating cell nuclear antigen of the rat thyroid gland before and after birth]. AB - We were studied the proliferative activity of the thyroid gland's cells of embryo and adult Wistar rats due to using the antiserum against the cell nuclear antigen (PCNA). The 100% of cells in thyroid's embryo was a positive on the 16th, 17th, 18th stages of the embryonic development (stages by Kornegy). The percent of PCNA positive cells considerably increased to 67% on the 19th stage. This fact the 20th and 21th stages of prenatal development relatively the previous stage coordinate with starting of the thyroid hormones in fetal thyroid gland and the first follicles formation. The small increasing of number of PCNA-positive cells detected on the 20th and 21th stages of prenatal development relatively the previous stage. Considerable elevation of the proliferating cells to 75% immediately before the birth (22th stage). An infant rats had have the 39% of proliferating cells. The 51% cells divided on the 5th day of postnatal development. Considerable decreased of the cell's division was occurred until the postnatal day 60. Using of the PCNA antiserum allowed to study cell proliferation in thyroid gland during pre- and postnatal rat development. PMID- 10457994 TI - [Biochemical parameters of the metabolism of the intercellular substance of connective tissue, correlating with morphometric signs of long bone lesions in an open aseptic fracture]. AB - The aim of the research was experimental study of dynamics and correlative dependences between biochemical connective tissue matrix metabolism indices (blood serum collagenase, cathepsin B, elastase, antielastase activity, hydroxyproline fractions and glycosaminoglycans concentration) and tissue damage morphometric indices after long bone aseptic osteotomy of rat in terms 3 h-60 days. The most strong and significant correlation was found between cathepsin B, elastase activity indices and dimensions of bone marrow ischemic damage focuses and summarised periosteal regenerates volume in bone fragments. PMID- 10457995 TI - [Effect of partial hepatectomy on the invertor mechanism of regulation of the Na+,K+-ATPase activity in hepatocytes of rats of various ages]. AB - Age peculiarities of partial hepatectomy effect on the hepatocytes plasma membrane Na+, K(+)-ATPase activity and its insulin-induced stimulation has been studied. It has been shown that partial hepatectomy does not change basal Na+, K(+)-ATPase activity in adult rats. In old partial hepatectomised rats Na+, K(+) ATPase activity is slightly higher than in control old rats, although this increase is not statistically significant. At the same time, partial hepatectomy acts differently on the insulin-induced Na+, K(+)-ATPase activation in adult and old rats. Insulin activates Na+, K(+)-ATPase at the same extent both in control and partial hepatectomized adult animals. In old hepatectomized rats, but not in old control animals, insulin stimulates Na+, K(+)-ATPase activity as well as. Thus hepatectomy "rejuvenates" old hepatocytes and results in recovery of invertor mechanism of Na+, K(+)-ATPase activation. PMID- 10457996 TI - [Features of the inhibition of catalase activity by 3-amino-1,2,4-triazole in erythrocytes and liver of rats with streptozotocin diabetes]. AB - Lipid peroxidation was intensified by streptozotocin induced diabetes mellitus in erythrocytes and liver. Activity of antioxidant enzyme superoxide-dismutase was decreased, activity of catalase was increased. Concentration of lipid peroxidation products was decreased after nicotinamide injections. It was investigated liver- and erythrocyte catalase inhibition in the presence of 3 amino-1,2,4-triazole. Effective inhibitor concentration for liver catalase by streptozotocin induced diabetes mellitus was 10 mM, by control-20 mM. Ascorbic acid induced catalase inhibition in the erythrocytes by diabetes mellitus increased by ascorbic acid concentration from 25 to 150 mM. [DHAA]/[AA]-ratio increased from 0.26 by control to 1.6 by diabetes mellitus and decreased to 0.44 after nicotinamide injections. PMID- 10457997 TI - [Catalase activity in the myocardium during stress in adult and aged rats]. AB - The study of catalase myocardium activity at stress in adult and old rats has been performed. It has been revealed that the formation of immobilization stress is accompanied by the enzyme activity stimulation. The expression of this effects is more significant in adult animals. Experimental data concerning the possible role of peroxidation lipids in myocardium catalase stress activation and its age modulation are given. PMID- 10457998 TI - [Effect of carnosine on the transformation of sodium nitrite in vitro in the presence of catalase]. AB - It was shown that carnosine changes the compacting of ferment's globe. On the one hand, that action is causes with the connecting of haem. On the other hand, carnosine influences on the hydrate's membrane around protein. Dipeptide influences also on the interaction of catalase with sodium nitrite, which intensity depends on sequence of introducing HNO2 and carnosine into the medium. PMID- 10458000 TI - [Effect of acute alcoholic and acetaldehyde poisoning on lipid metabolism in the rat liver]. AB - Lipid metabolism in the liver of rats in acute alcohol (25% solution, intraperitoneally, 4 g/kg, 30 minutes) and acetaldehyde (5% solution, intraperitoneally, 0.266 g/kg, 30 minutes) intoxication has been studied. It has been revealed that with acute injection of ethanol into the livers of experimental animals the level of cholesterol is decreased, the content of triacylglycerols and phosphatidylethanolamine is increased. Analogous changes in the concentration of lipid fractions have been also revealed after injection of acetaldehyde. PMID- 10457999 TI - [Content of fat-soluble vitamins in human blood serum during the use of food additives with vitamin E and carotene of plant origin]. AB - Vitamin E, A and carotene levels in blood serum of human that were affected by the definite irradiation doses accepting the admixtures of vitamin E and carotene preparations were estimated. The consuming of vitamin E increases vitamin E and A content in blood serum. While obtaining carotene significant increase of vitamin E and carotene content is observed, but it does not influence to vitamin A content. Simultaneous application of both preparations leads to vitamin E level growth being similar to persons, receiving only vitamin E; in this case the vitamin A and carotene content is increasing more actively and produces as early as in the month the carotene accumulation in blood serum. Along side with this the vitamin A content increases only in 3 month, and vitamin E practically does not change. Among estimated persons 45 years older revealed content of vitamin E increase in 3 month of both application of preparation. Application of vitamin E and carotene preparations have expressive positive change of fat-soluble vitamin status in blood serum. PMID- 10458001 TI - [Effect of the biopreparation "Polyene" on parameters of blood fatty acid composition in volleyball athletes]. AB - Influence of the preparation "Polyen" has been studied on indexes of lipid acids contents in blood plasma, erythrocyte membrane and serum albumin of high qualified volleyball-players. It was shown that "Polyen" has favourable effect on different metabolic pathways of lipid acids, that allow to recommend this preparation as additional component in food ration for training sportsmen. PMID- 10458002 TI - [Combined action of xenobiotics on the status of the antioxidant system of the body]. AB - The chronic combined inhalational effect of lead, radon, nitric oxides (NO, NO2) and amorphous hydrophobic silicon dioxide on the activity of enzymes such as superoxide dismutase, catalase, glutathione peroxidase, glutathione-s transferase, g-glutamyl transpeptidase, peroxidases and also on the protein reducing glutathione concentration of the mongrel male-rat was investigated. It has been shown that organism antioxidant system displaces activity toward of the most widespread xenobiotics, expressing the rising of itself activity during whole experience. The dynamic of changes of the antioxidant system activity also was analyzed. Moreover, at the first step of our experience there was the sharp activity rise as a result of action of the mechanisms of adaptation and compensation. At the following intermediate step we were observing exhaustion of the antioxidant system and including of additional powers at the last step. PMID- 10458003 TI - [Effect of gamma-aminobutyric acid on Mg2+-ATPase of brain microsomes of the fish Abramis brama L]. AB - The influence of the GABA on the Mg(2+)-ATPase from microsomal fraction of fish brain (Abramis brama L.) was investigation. Preincubation of the microsomes with different concentration of GABA (10(-8)-10(-4) M) stimulated Mg(2+)-ATPase activity. This effect of neuromediator is sensitive to picrotoxine (10(-4) M). It was established that Mg(2+)-ATPase activity stimulated by anions (Cl > Br > F), inhibited by SCN- and not effected by HCO3-. The influence of the anions on the Mg(2+)-ATPase is liable to be inhibited by picrotoxine. It was supposed that anion-sensitive Mg(2+)-ATPase is associated functionally with GABAa-receptor. PMID- 10458004 TI - [The problem of determining the affinity of bivalent antibodies using ELISA and its mathematical solution]. AB - A new approach for the determination of the affinity of monovalent receptors in direct coordinates could be obtained without using Klotz or Scatchard equations. The mathematical solution of the problem of the determination of the affinity of bivalent antibodies by ELISA gives the following relation between absorbance, measured in ELISA. and antigen concentration: omega + square root of omega 2 + omega = liK, where omega = (Ao-Ai)/Ai, Ai is the absorbance, if Ag concentration is equal li; Ao is the absorbance, if Ag concentration is equal zero, K is an equilibrium constant. PMID- 10458005 TI - [Comparative characteristics of various methods for determining the kinetic parameters of first-order reversible and irreversible reactions]. AB - The two methods to determine a rate constant and a total reaction product for both reversible and irreversible reaction, using the data of reaction product kinetics, are proposed. The methods are based on analytical solution of the system of transcendental equations (for special case), which describes such processes. It was shown that these methods are precise and are more simple and convenient than the methods proposed by other authors. PMID- 10458006 TI - Ultrasound secretin test in patients with pancreas divisum--an aid in the diagnosis of papillary or dorsal duct stenosis? AB - BACKGROUND: Pancreas divisum is a common anatomical variant of pancreatic ductal anatomy. Obstruction of the accessory papilla could cause pain and pancreatitis. It has been suggested that accessory papillary sphincter obstruction can be assessed by sonographic measurement of pancreatic duct diameter after secretin stimulation. METHODS: We now compared our results of sonographic pancreatic duct diameter measurements before and during 10 min after intravenous injection of 1 CU secretin per kg body weight in 32 patients with confirmed pancreas divisum and 20 healthy volunteers. RESULTS: The healthy controls showed a short-lasting duct caliber enlargement by about 93% of the basal diameter within 5 min after secretin injection. 25 pancreas divisum patients without pancreatic disease had a secretin-induced duct dilatation by about only 58%. In four patients with pancreas divisum and chronic pancreatitis no or just a slight duct dilatation was observed after stimulation. Two patients with dorsal duct stenosis as well as one patient with accessory papilla stenosis, however, showed a marked and prolonged secretin-induced duct enlargement by about 155% of the basal duct diameter. CONCLUSION: In this investigation pancreatic duct response to secretin stimulation in pancreas divisum patients without pancreatic disease was less marked than in normal individuals. Thus, a particularly distinct and long-lasting duct dilatation could support the suspicion of accessory papilla or pancreatic duct stenosis. PMID- 10458007 TI - [Propofol for sedation in gastroscopy--a randomized comparison with midazolam]. AB - Midazolam, a benzodiazepine with amnestic and sedative effects is the drug of choice for sedation of patients undergoing upper gastrointestinal endoscopy. Propofol, a phenolic derivate, is a short-acting anesthetic producing a more rapid onset sedation amnesia and a shorter recovery than midazolam: In higher doses it acts as hypnotic. The aim of this study was to evaluate both drugs in a prospective randomized trial for sedation of patients undergoing esophagogastroduodenoscopy (EGD). METHODS: 60 patients undergoing EGD were randomized to receive either propofol (n = 30) or midazolam (n = 30). No other analgetic or sedative drugs were used. 3 l oxygen were given routinely by nose. Blood pressure, oxygen saturation (pulse oxymetry) and heart rate was monitored continuously. The sedation quality was determined from the endoscopist and patient separately by use of a scale as either good, fair or insufficient. RESULTS: Changes of the heart rate and oxygen saturation showed no differences in both groups (> 0.05). The blood pressure decreased significant by using propofol (p < 0.01). The sedation quality was good in both groups without significant differences. The recovery time was shorter if propofol was administered (p < 0.01). CONCLUSIONS: Propofol is an alternative drug for sedation in upper endoscopy. It showed same sedation quality as midazolam with the advantage of a short recovery time. Because of a possible decrease of the blood pressure continuous monitoring is recommended. PMID- 10458008 TI - Fecal excretion of alpha 2-macroglobulin: a novel marker for disease activity in patients with inflammatory bowel disease. AB - BACKGROUND: Quantification of fecal alpha 1-antitrypsin (AAT) excretion is established for estimation of enteric protein loss and assessment of disease activity in inflammatory bowel disease (IBD). In contrast, little is known about prevalence, course, and clinical significance of intestinal leakage of larger size serum antiproteinases in these disorders. SUBJECTS AND METHODS: Firstly, 23 IBD patients (Crohn's disease, CD, n = 17, and ulcerative colitis, UC, n = 6) were examined at 34 independent episodes (relapse, n = 16, remission, n = 18) for parallel serum and fecal alpha 2-macroglobulin (AMG) and AAT concentrations by standard immunonephelometry, and compared to 40 healthy controls. From these IBD patients, secondly, a random cohort of twelve individuals (9 CD, 3 UC) was prospectively followed for those parameters at about monthly intervals for 7-14 (median 10.5) months. RESULTS: The threshold of detection for fecal AMG concentration was about 0.06 mg per gram dry weight stool (mg/g dws) under the present analytical conditions. While in healthy subjects fecal AMG was demonstrated at very low levels only (< or = 0.07 mg/g dws), it was found in CD and UC patients at elevated concentrations of < 0.06-3.18 (median 0.17) and < 0.06-1.91 (median 0.40) mg/g dws, respectively. Fecal AMG contents were more increased in active IBD compared to quiescent disease (p = 0.03), and they correlated to Crohn's Disease Activity Index in CD patients (p = 0.05), while not to Clinical Activity Index in UC individuals (p = 0.46). Post hoc evaluation of follow-up data suggested two distinct groups of IBD patients either with or without consistently detectable fecal AMG excretion, with the first ones exhibiting a more active clinical course than the latter ones (p < or = 0.02). CONCLUSIONS: AMG is excreted in feces of healthy subjects in traces only, while its stool concentration is largely increased in IBD patients where it reflects clinical disease activity. This novel stool parameter may be of potential value in the diagnostic and prognostic management of these individuals. PMID- 10458009 TI - Diagnosis of Chilaiditi's syndrome with abdominal ultrasound. AB - Chilaiditi's sign is a radiographic term used when the hepatic flexure of the colon is seen interposed between the liver and right hemidiaphragm. When symptomatic, this is Chilaiditi's syndrome. We report a case of a 70-year-old man who presented with abdominal pain, vomiting, singultus and constipation. Ultrasound was initially performed which showed an intestinal loop between the anterior surface of the right liver lobe and the diaphragm. The chest X-ray revealed colon gas under the right diaphragma and the abdominal CT-scan confirmed the hepatodiaphragmatic interposition of the colon. Colonic elongation and laxity of colonic and hepatic suspensory ligaments are the principal predisposing factors. The advantages of the abdominal ultrasound in the diagnosis and follow up as well as possible complications and forms of therapy with this syndrome are discussed. PMID- 10458010 TI - Malabsorption with progressive weight loss and multiple intestinal ulcers in a patient with T-cell lymphoma. AB - We describe a 52-year-old woman who presented with severe diarrhea, nausea, intermittent abdominal pain and weight loss of 18 kg within ten months. Jejunal and duodenal ulcers were detected by endoscopy and multiple biopsies revealed villous atrophy of the jejunum. However, neither gliadin nor endomysium antibodies were detected and no clinical and histological improvement was achieved after gluten withdrawal. Despite strong clinical suspicion for intestinal lymphoma many unrevealing biopsies were done. The patient developed intermittent septic fever and diagnostic laparotomy revealed jejunal perforation. Partial jejunal resection was performed and histology confirmed the diagnosis of an intestinal T-cell lymphoma without celiac disease. Malabsorption and all intestinal ulcers disappeared during the course of chemotherapy (six cycles CHOP) and the patient recovered remarkably. PMID- 10458012 TI - [Methotrexate in therapy of gastrointestinal diseases]. AB - This review is not only a comprehensive synopsis of the published data on methotrexate treatment for inflammatory bowel disease and primary biliary cirrhosis but gives also an overview about acting mechanisms, pharmacologic properties and limiting toxicities of low-dose methotrexate. Methotrexate and its polyglutamate analogues are structurally related to folic acid and inhibit many enzymes in the metabolic pathway of folic acid. Long-term low-dose methotrexate treatment (7.5-25 mg) inhibits production of thymidylate, purines, and methionine and leads to intracellulary accumulation of adenosin. These actions lead to inhibition of cellular proliferation, decreased formation of antibodies and reduced productions of inflammatory medicators. Several trials have demonstrated the efficacy of low-dose methotrexate therapy for certain indications as a promising new agent in the treatment of inflammatory bowel disease and primary biliary cirrhosis. However, several serious toxicities potentially limit its use. PMID- 10458011 TI - Therapy of pseudomyxoma peritonei of appendiceal origin--surgical resection and intraperitoneal chemotherapy. AB - Pseudomyxoma peritonei (PMP) is rare clinical entity presenting with mucus producing lesions on the peritoneal surface. PMP is found in middle-aged or older patients in approximately two out of 10,000 laparotomies. Most of the reported cases are due to a primary process located in either appendix or the ovaries. PMP presents with an insidious onset of symptoms, and is characterized by long-term survival with good general health and absence of visceral invasion or distant metastasis. The treatment of choice for PMP is surgical resection and the removal of free mucus. Despite a high rate of disease recurrence, adjuvant or palliative chemotherapy has not yet been clearly established. In our opinion, intraperitoneal application may be more effective than a systemic regimen because only local treatment can provide sufficient concentrations of chemotherapeutic agents. In view of the lack of standardized treatment regimens and based on our experience, we recommend intraperitoneal chemotherapy with 5-FU and mitomycin C or cisplatin. We report ten cases of PMP of appendiceal origin and present a review of the literature on this disease and its treatment. PMID- 10458013 TI - [New molecular aspects of cholestatic liver diseases]. AB - Hepatic uptake and biliary excretion of bile salts and non-bile salt organic anions (e.g., bilirubin) is mediated by specific transport proteins located at the basolateral and canalicular membranes of hepatocytes. Several hepatobiliary transport systems have been identified and cloned over the past years. This development has facilitated molecular biological and genetic analyses of these transporters in experimental cholestasis and human cholestatic liver diseases. Evidence now exists that decreased or even absent expression of hepatobiliary transport systems may explain impaired transport function resulting in hyperbilirubinemia and cholestasis. This review summarizes the molecular defects in hepatocellular membrane transporters associated with hereditary and acquired forms of cholestatic liver diseases. The increasing information on the molecular regulation of hepatobiliary transport systems should bring new insights into the pathophysiology and treatment of human cholestatic liver diseases. PMID- 10458014 TI - [Is lack of stomach carcinoma prevention by Helicobacter pylori eradication unethical?]. PMID- 10458015 TI - [Interferon-alpha plus ribavirin combination therapy in chronic hepatitis C]. PMID- 10458017 TI - [Noninvasive detection of colorectal carcinomas by analysis of expression of CD44 variant coded transcripts]. PMID- 10458018 TI - [The pulmonary arteries and veins of miniature swine, together with a common view of their supportive and stabilizing functions in the lung]. AB - With respect to distribution and flow, the Vasa pulmonalia resemble those of the normal grown domestic pig to a great extent. The present article investigates whether, like the human lung (Hayek, 1970), the arrangement of the blood vessels has a function over and above that of directing blood; that is to say, a mechanical function. The arteries through which the blood streams act as flexible supporting structures, and the veins as elastic holding structures, for the quadrupede animal also. PMID- 10458016 TI - [Cholysarkosin in treatment of bile acid deficiency syndrome]. PMID- 10458019 TI - [Structure of infrapatellar synovial ligament in dogs]. AB - The knee joint of the dog is frequently the subject of surgery if the cruciate ligaments or the menisci are injured. In the knee joint of the dog we find a ligament, which is spread out between the anterior part of the fossa intercondylaris ossis femoris and the corpus adiposus infrapatellaris. This ligamentum synoviale infrapatellare has not previously been described and represents a homologous structure to the plica synovialis infrapatellaris of human knee joint. Just like the latter and just like the cruciate ligaments it is a rudiment of a septum articulare genu, which is spread out between an articulatio femorotibialis and an articulatio femorofibularis in the early phylogenesis of tetrapods. The ligamentum synoviale infrapatellare of the dog consists of a stratum synoviale and a stratum spinosum, which includes blood vessels, nerves, many collagenous fibres and some elastic fibres. We do not find evidence for sensory nerve endings or corpuscular receptors. Therefore a servering of the ligament during knee joint surgery does not affect the proprioceptive potential of the knee joint. PMID- 10458020 TI - Angiogenesis in the bovine corpus luteum: an immunocytochemical and ultrastructural study. AB - Immediately after ovulation a neovascular response occurs at the level of the theca interna. Pericytes and endothelial cells of post-capillary venules locally remodel the surrounding stroma, elongate and migrate into the avascular granulosa folds of the ruptured follicle. In order to examine the composition of the extracellular matrix as well as the growth characteristics of these newly formed vessels, we used immunohistochemical and electron microscopic methods. Initial sprouts were characterized by the appearance of a fibrillary network of fibronectin along the main axis of the sprout. Type IV collagen stained weakly and extracellular deposits of laminin were amorphous and patchy around immature capillary sprouts. In advanced maturational stages of the sprouts the capillaries were surrounded by increased deposits of fibronectin, whereas laminin and type IV collagen displayed a distinct and well-developed line around endothelial cells and pericytes. These observations indicate that the microvascular extracellular matrix undergoes a series of quantitative rather than qualitative changes during capillary development before achieving final maturation. Ultrastructural analyses showed that early capillary sprouts in the bovine corpus luteum were usually preceded by pericytes migrating at the tips of the sprouts. Endothelial cells comigrated in cohesive cylindrical projections, forming immediately a slit-like lumen which satisfies the criteria of the intercellular canalization type. Pericytes at the tips of endothelial sprouts exhibited a slender, bipolar morphology and were regularly surrounded by fragmented basal lamina, which is well-developed around pericytes in a more proximal position of the sprout. The regular association of pericytes with the leading front of the capillary sprouts suggests that these cell types may serve as guiding structures aiding outgrowth of endothelial cells in the bovine corpus luteum. PMID- 10458021 TI - Arteries of the carotid body in rats. AB - The arterial blood supply of 40 carotid bodies in 20 Wistar rats of both sexes, aged 10-12 weeks (250-350 g), was examined by light microscopy. The carotid bodies of all rats were supplied by only one carotid body artery. The average diameter of the carotid body arteries was 40 microns. This artery arose either from the external carotid artery (97.5%) or the occipital artery (2.5%). There was an intimal cushion at the origin of the carotid body artery. The carotid body artery, after reaching the caudal pole of the carotid body, divided into the first-order branch. In the carotid body, the paranchyma was divided into the second-order branch. The carotid body artery was of the muscular type. PMID- 10458022 TI - Characterization and distribution of B cells in the lymphoid organs of goats. AB - The distribution of B cells in the lymphoid organs of the goat was studied using a panel of 13 monoclonal antibodies (mAbs) developed against different markers for bovine B cells. Samples of mesenteric lymph nodes, jejunal and ileal Peyer's patches, duodenum, jejunum, ileum, colon, caecum and rectum were taken from four 7-month-old male Murciano-granadina goats using the avidin-biotin-peroxidase (ABC) method on frozen sections as described by Hsu et al. (1981). The mAbs against immunoglobulins (Ig) recognized a large number of cells, particularly in the light zones of the germinative centres of the lymphoid follicles, regardless of the isotype against which they were directed. However, the greatest numbers of B cells in the germinative centres and outer coronas of the lymphoid follicles of the lymph node, spleen and Peyer's patches were recognized by mAbs against the L lambda chain of Ig and against IgM. This was also the case in other locations where B cells were abundant, such as the medulla of the lymph node and the dome of the Peyer's patches. These mAbs recognized not only B lymphocytes but also plasma cells, showing an intracytoplasmatic reaction (numerous in the spleen red pulp and the intestinal lamina propria when mAbs were used against the L lambda chain of the Ig, scarce in the intestinal lamina propria when used against IgM and scarce in spleen red pulp and numerous in the intestinal lamina propria when mAbs against IgA were used). The mAbs BAQ44 A, GC65 A and GB25 A are of interest because, besides marking cells in the B areas where lymphocytes show surface Ig, they give a positive reaction in areas where there are Ig-cells (the dark zone of the germinative centre) and do not immunostain plasma cells. Thus, these mAbs recognize a surface marker which is not an Ig. PMID- 10458023 TI - [Anatomic features of the carpal joint of the cheetah (Acinonyx jubatuw), compared with the domestic cat (Felis catus)]. AB - The anatomy of the carpal joint of the cheetah (Acinonyx jubatus) was examined in seven specimens using dissection and corrosion casts as well as radiography, and compared to well-known data of the domestic cat (Felis catus). It was found that in the cheetah, as in the domestic cat, the intermedioradial, ulnar and accessory carpal bones, as well as the first, second, third and fourth carpal bones and the sesamoid bone of the abductor pollicis longus muscle, develop in a regular manner. The bones had a similar shape and the ligamentous apparatus was comparable, the most striking differences being the connection of all compartments of the joint cavity and the mediocarpal joint, working as a screw joint. The syndesmosis between the intermedioradial and ulnar carpal bones, instead of a synovial connection, is another adaptation for stabilization of the carpus of the cheetah during locomotion. The joint capsule is little spacious and in all three recesses can be differentiated. The first extends proximally palmar the ulnar carpal bone between the styloid process of the ulna and the accessory carpal bone, the second also extends proximally mediopalmar of the intermedioradial bone, and the largest third recess is located on the dorsal surface and extends proximally, laterally to the inserting tendon of the extensor carpi radialis muscle. PMID- 10458024 TI - Morphological characterization of gland cells of the glandular sac area in the complex stomach of the Bactrian camel (Camelus bactrianus). AB - The morphology of the gland cells in the glandular sac area of the Bactrian camel and the composition of secretory substances were examined by histochemical methods. It was found that the gland cells of the glandular sac area were of the same type and size as those of the cardiac glands. The composition of secretory substances from the glandular sac area was the same as that of secretory substances from the cardiac glands. Moreover, secretory substances from the gland cells of the glandular sac area contained a great deal of acid glycoconjugates, such as sialic acid, in addition to neutral saccharides (fucose, mannose, glucose, N-acetyl-D-glucosamin, galactose and N-acetyl-galactosamin). Furthermore, immunohistochemical examination showed that progastricsin was present in the gland cells of the glandular sac area and the cardiac gland. In this study, histological analysis suggested that the stomach of the Bactrian camel is a single cavity stomach, formed as a result of multiple differentiation and growth of cardiac glands through the process of evolution. PMID- 10458025 TI - Association between supraependymal nerve fibres and the ependymal cilia of the mammalian brain. AB - The ciliated ependymal cells that line the floor of the fourth ventricle in adult Wistar rats are overlaid by an immense network of supraependymal nerve fibres. This study presents morphological evidence of a characteristic association of supraependymal nerve fibres with the cilia of the ependymocytes of the ventricular surface. It is found that the supraependymal nerve fibres accumulate and/or terminate at the base of the clusters of ependymal cilia. These fibres are varicose in nature and consist of large-diameter fibres with numerous prominent varicosities and small-diameter fibres with less frequent and less prominent varicosities. The characteristic association of the supraependymal nerve fibres with the ciliated ependymocytes suggests that these nerve fibres may be involved in producing co-ordinated ciliary movements in the mammalian brain. PMID- 10458026 TI - Localization of the N-methyl-D-aspartate R1 receptor in the pituitary gland of immature rainbow trout (Oncorhynchus mykiss). AB - A study was conducted on the selective immunostaining of pituitary cells and pars nervosa of immature rainbow trout by antibody raised against the R1 subunit of the rat ionotropic N-methyl-D-aspartate (NMDA) receptor. In the pars distalis, the mammotrophs of the rostral pars distalis exhibited the most marked imunoreactivity, with the somatotrophs of the proximal pars distalis showing a consistently lower degree of immunoreactivity. No imunoreactivity was associated with the corticotrophs, thyrotrophs or gonadotrophs. In the pars intermedia, the melanotrophs showed no evidence of immunoreactivity, whereas the putative somatomammotrophs exhibited a wide inter-animal range of immunoreactivity. Some imunostaining was also evident in the anterior and posterior part of the pars nervosa, at the interface of the pars distalis or pars intermedia with the pars nervosa. PMID- 10458027 TI - Histological characterization of defective spermatogenesis in mice lacking the basigin gene. AB - Basigin is a transmembrane protein belonging to the immunoglobulin superfamily. In the light of the fact that knockout mice lacking the basigin gene (Bsg) are azoospermic, the phenotype in the male reproductive system was extensively examined in this study. Spermatogenesis in Bsg (-/-) mice was found to be disrupted, and arrested at the metaphase of the first meiotic division. A few germ cells differentiated into young spermatids, but they were exfoliated. The lumens of the male reproductive system were filled with round degenerated cells. Using the TUNEL method and electron microscopy, some of the degenerated cells in the testis and epididymal head were shown to be apoptotic. Crystalloids of fine tubules and unusual ectoplasmic specializations were also observed in the Sertoli cells of Bsg (-/-) mice. These specializations displayed unusual 'circular' structures. Furthermore, unusual ectoplasmic specializations covering the spermatocytes rather than the mature spermatids were found. These structures were formed as a result of the lack of mature spermatids in the Bsg (-/-) testis. Results from analyses of azoospermia in the Bsg (-/-) mice suggest that basigin, through the interactions between germ cells and Sertoli cells, is an essential factor in the growth and/or survival of spermatids. PMID- 10458029 TI - [Oral dormicum premedication of children in day surgery]. AB - Thirty-four patients aged 5-14 years were observed, divided into age groups OF 5 8 and 8-14 years. Cardiovascular function was evaluated by central hemodynamic parameters and cardiointervalography. Changes in external respiration parameters were studied. The efficacy of premedication was assessed by objective parameters and by clinical signs. Each patient was examined 4 times at the following stages: 1) before premedication; 2) at the 10th min; 3) at the 20th min; and 4) at the 30th min. Oral premedication with dormicum in a dose of 0.5 mg/kg did not affect heart rate or arterial pressure. By the 20th min it notably decreased total peripheral resistance, significantly decreased the respiratory volume, increased respiration rate, and decreased sympathetic tone in younger children. In older children the premedication virtually did not affect total peripheral resistance and facilitated heart work, increased the respiratory volume, rarefied the respiratory rate, and did not affect the autonomic regulation balance. Younger children fell asleep without excitation or negativism; 35.2% developed psychomotor reactions. In older group the effect was good in 71.1% children, the rest had "apparitions" starting from the 17th min: doubling of the staff's faces, white spiders, or white moths. PMID- 10458028 TI - [Clinical experience in the use of clonidine in regional anesthesia and early postoperative period (1984-1989)]. AB - Clonidine attracts attention as a sole agent for local anesthesia. This review analyzes the role of alpha 2-adrenergic agents in modern regional anesthesiology, both practical and experimental. Recent controlled studies showed that an alpha 2 adrenoreceptor clonidine administered spinally is characterized by a dose dependent antinociceptive effect and is as effective as epinephrine in prolonging local anesthetic blocks. Moreover, in combination with spinal opioids, intraspinal clonidine possesses a synergic analgesic effect, potentiating the intensity and duration of opioid anesthesia. Use of clonidine as an adjuvant to various plexus blockades and for caudal block in children is discussed. Regional administration of clonidine involves side effects, such as hypotension, bradycardia, and sedation. Minimization of the dose of alpha 2-adrenergic agonists minimizes the complications. Regional clonidine effects (analgesia, hemodynamic changes, and sedation) and their recognized mechanisms of action are described. PMID- 10458030 TI - [Spirometric evaluation of external respiration function in laparoscopic and mini laparotomy cholecystectomy]. AB - Spirometric parameters are studied in 54 patients subjected to laparoscopic cholecystectomy and cholecystectomy through a mini-laparotomy approach. The restrictive and obstructive disorders in the external respiratory function after minilaparotomy cholecystectomy are less expressed than after laparoscopic cholecystectomy. Thorough examination of the external respiratory function is recommended when defining the indications for laparoscopic cholecystectomy. PMID- 10458031 TI - [Choice of drugs for general anesthesia in patients with arrhythmia]. PMID- 10458032 TI - [Estimation, control and assessment of the anesthesiologists' work]. AB - A scheme for assessing anesthesiologist's work is proposed, based on qualimetric principles and developed using qualimetric methods, the basic of which is comparative quantitative analysis of actual work and norm-setting, standard, and mean parameters. The work of an anesthesiologist is evaluated by quantity, difficulty, and quality. Methodological work and consultations are taken into account. Coefficients characterizing the deviation of actual work from the standard are calculated for each sign. An integral coefficient is estimated for unambiguous evaluation. The proposed score system is rather simple and helps more objectively evaluate the work of an anesthesiologist. PMID- 10458033 TI - [Diagnosis and treatment of patients with postintubation stenosis of the larynx and trachea]. AB - Experience gained in the diagnosis and treatment of 498 patients with postintubation laryngotracheal stenosis is summarized. Use of accessory methods of diagnosis, such as x-ray tomography of the trachea and mediastinum and endofibroscopy of the larynx and trachea is validated. Flexible endoscopes with kits of special instruments allow decannulation without subsequent therapy in 95% patients. Simultaneous tracheoplasty and retracheostomy simplify surgical treatment of patients with postintubation laryngotracheal stenosis, making it shorter by 1-2 stages. The development of laryngotracheal stenoses depends on the duration of intubation, and hence, the optimal term for tracheostomy in patients on forced ventilation of the lungs is day 3 after intubation. PMID- 10458035 TI - [State of the hemostasis and fibrinolysis systems in cesarean section under combined ketamine anesthesia]. AB - The reserve potentialities of the hemostasis and fibrinolysis systems were studied by thromboelastography in 34 women with normal pregnancy aged 18-34 years at different stages of cesarean section under combined ketamine anesthesia. Three types of responses were observed: compensated, subcompensated, and decompensated. In women with subcompensated response removal of the fetus from the uterus involved in 30.1% cases depletion of the reserve potential of anticoagulant mechanisms, which led to abnormal blood loss in 34.7% cases. PMID- 10458034 TI - [Latex allergy: a current medical problem]. PMID- 10458036 TI - [Clinico-pathogenetic variants of DIC syndrome in patients with severe craniocerebral trauma]. AB - Detection of clinical and pathogenetic variants of the DIC syndrome for development of its differentiated therapy in multiple-modality treatment of severe craniocerebral injury was the purpose of this study. A total of 170 patients with grave craniocerebral injury were examined. The hemostasis system was studied by the following methods: analysis of platelet hemostasis, general coagulation tests, fibrinolysis evaluation, detection of physiological anticoagulants and markers of intravascular blood coagulation and fibrinolysis. Based on the clinical (intra- and extracranial) symptoms and results of studies of the hemostasis system, 3 clinical pathogenetic variants of the DIC syndrome were distinguished, which should be borne in mine when treating patients with severe craniocerebral injury developing the DIC syndrome. PMID- 10458037 TI - [Nosocomial pneumonia in intensive care units (review of the literature)]. PMID- 10458038 TI - [Hospital infections caused by Pseudomonas aeruginosa. Significance in intensive therapy]. AB - The significance of P. aeruginosa as an agent of hospital infections in intensive care departments is determined by high prevalence of this microorganism, its natural and acquired resistance to antibiotics of various groups, and severity of the infection it induces. The resistance of P. aeruginosa to antibiotics is different in different regions. Among the strains isolated in Moscow in intensive care wards for newborns 9% were resistant to meropenem, 10% to amicacine, 15% to imipramine, 16% to cefepime, 37% to ceftasidime, 45% to piperacylline/tasobactam, 45% to ciprofloxacine, and 60% to gentamicin; 1.5% of these strains were resistant to all tested antibiotics. High prevalence of antibiotic resistance among P. aeruginosa impedes the choice of drugs for empirical antibiotic therapy and increases the significance of microbiological diagnosis. Even if an agent is sensitive to such antibiotics as semisynthetic penicillines and aminoglycosides, their use as monotherapy in infections caused by P. aeruginosa is ineffective. Carbapenemes, III- IV generations cefalosporines, and fluoroquinolones can be used as mono therapy. PMID- 10458039 TI - [Problems and perspective approaches to correction of mediator response in sepsis]. PMID- 10458040 TI - [Use of octagam and pentaglobin in the treatment of severe sepsis in the neonatal period]. AB - Many patients with sepsis are immunocompetent. They respond well to standard combinations of antibiotics and cardiovascular support. However, immunocompetent patients, particularly small-for-date newborns, are in need of stronger immune protection. Although the mechanisms due to which antibody preparations yield positive effects are not quite clear, clinical studies carried out in many countries and our own results indicate a high protective effect of intravenous immunoglobulins and necessitate their use at least in intensive care departments. PMID- 10458041 TI - [Present-day concepts concerning immunoprophylaxis and immunotherapy of surgical infections]. PMID- 10458042 TI - [Monitoring accuracy of ventilation parameters and gas mixture composition during anesthesia and artificial ventilation of the lungs]. PMID- 10458043 TI - [Case of successful treatment of traumatic hemopericardium by puncture drainage in a resuscitation and intensive care unit of a municipal hospital]. PMID- 10458044 TI - [Replenishment of circulating blood volume by the use of colloid solutions]. PMID- 10458045 TI - Perfectionism and self-consciousness in social phobia and panic disorder with agoraphobia. AB - Social phobics were compared to patients with panic disorder with agoraphobia and normal controls on perfectionism and self-consciousness. On concern over mistakes and doubts about action, social phobics scored higher than patients with panic disorder. Social phobics also demonstrated a higher level of public self consciousness than patients with panic disorder and when this difference was controlled for the significant differences on perfectionism disappeared. Within each patient group, however, perfectionism was more robustly related to social anxiety than was public self-consciousness, which replicates the findings of Saboonchi and Lundh [Saboonchi, F. & Lundh, L. G. (1997). Perfectionism, self consciousness and anxiety. Personality and Individual Differences, 22, 921-928.] from a non-clinical sample. The results are discussed in terms of public self consciousness being a differentiating characteristic of the more severe kind of social anxiety which is typical of social phobia. PMID- 10458046 TI - Cognitive factors involved in the onset and maintenance of posttraumatic stress disorder (PTSD) after physical or sexual assault. AB - Cognitive factors hypothesised to influence the development and maintenance of PTSD were investigated. 92 assault victims completed questionnaires assessing a range of cognitive variables. Factors relating to onset of PTSD were investigated by comparing victims who did and who did not suffer PTSD. Factors relating to maintenance of PTSD were investigated by comparing victims who had recovered from PTSD with victims who had persistent PTSD. Cognitive factors associated with both onset and maintenance of PTSD were: appraisal of aspects of the assault itself (mental defeat, mental confusion, appraisal of emotions); appraisal of the sequelae of the assault (appraisal of symptoms, perceived negative responses of others, permanent change); dysfunctional strategies (avoidance/safety seeking) and global beliefs impacted by assault. Cognitive factors that were associated only with the onset of PTSD were: detachment during assault; failure to perceive positive responses from others and mental undoing. Relationships between the cognitive variables and PTSD remained significant when variations in perceived and objective assault severity were statistically controlled. The cognitive factors identified in the study may contribute to PTSD directly, by generating a sense of ongoing threat, or indirectly, by motivating cognitive and behavioural strategies that prevent recovery, or by affecting the nature of the traumatic memory. PMID- 10458047 TI - Frequency and comorbidity of social phobia and social fears in adolescents. AB - This report presents findings on the frequency, comorbidity and psychosocial impairment of social phobia and social fears among 1035 adolescents, aged 12-17 years. The adolescents were randomly selected from 36 schools in the province of Bremen, Germany. Social phobia and other psychiatric disorders were coded based on DSM-IV criteria using the computerized Munich version of the Composite International Diagnostic Interview. Seventeen (1.6%) of the adolescents met the DSM-IV criteria for social phobia sometimes in their life. More girls than boys received the diagnosis of social phobia and the frequency of the disorder increased with age. The lifetime frequency of social fears were much higher than that of social phobia. The most common types of feared social situations were fear of doing something in front of other people, followed by public speaking. Social phobia comorbid highly with depressive disorders, somatoform disorders and substance use disorders. Despite the high level of psychosocial impairment experienced by cases with social phobia and those with any social fears, only a small portion of them did receive professional help. PMID- 10458048 TI - Context-specificity of relapse: effects of therapist and environmental context on return of fear. AB - Context-specificity of fear extinction was tested among 65 participants who were fearful of spiders by manipulating the contexts used for exposure treatment and two-week follow-up assessment. Context was defined by both meaningful (presence of a particular therapist) and incidental (room location and furnishings) environmental cues. Distinct phobic stimuli were used to examine interactions of context with stimulus. Physiological, behavioral and verbal indices of fear were measured. Results provided modest support for context-specific return of fear. With one stimulus, participants assessed in a non-treatment context at follow-up exhibited greater returns in heart rate levels. In addition, three of four participants who could not touch the stimulus at follow-up had been tested in a non-treatment context. Future investigations may benefit from greater distinctions between contexts or manipulation of contextual features more directly relevant to fear. Finally, post hoc analyses identified high trait anxiety, slow treatment response, recovery of phobic cognitions and long duration/high intensity phobic encounters post-treatment as significant predictors of increased return of fear. PMID- 10458049 TI - The structure of specific phobia symptoms among children and adolescents. AB - Previous research [Frederikson, M., Annas, P., Fisher, H. & Wik, G. (1996). Gender and age differences in the prevalence of specific fears and phobias. Behaviour Research and Therapy, 34, 33-39.] has shown that specific phobia symptoms of adults cluster into three subtypes: animal phobia, blood-injection injury phobia and environmental-situational phobia. The present study examined whether these specific phobia subtypes can also be found in children. 996 children aged between 7 and 19 years completed a brief questionnaire regarding the frequency with which they experienced specific phobia symptoms. Confirmatory factor analysis was employed to examine the structure of these data. Results showed that childhood specific phobia symptoms indeed cluster into the three subtypes as described by Frederikson et al. and that these subtypes are either intercorrelated or the product of a single higher order factor. This structure appeared to be largely invariant across genders and age groups. PMID- 10458050 TI - Caffeine effects on mood and memory. AB - The purpose of the present research was to assess whether a psychoactive dose of caffeine would have differential affects on the mood dimensions of arousal versus feelings of pleasantness and whether these mood alterations would influence memory either by (1) the experience of arousal at learning and/or (2) altered and congruent mood states at learning and recall. To address these questions, the administration of 5 mg/kg caffeine or placebo at learning and retrieval sessions was manipulated and subjects' mood was evaluated by several different self-report measures. Sixteen words were incidentally studied during the learning session and memory was evaluated by the number of words correctly recalled at the retrieval session two days later. Results revealed that caffeine reliably increased arousal, but did not affect any emotion dimensions related to feelings of pleasure. Subjects who received caffeine at learning and retrieval were also in equivalent mood states at both sessions. Moreover, caffeine did not produce any effects on memory; thus, neither hypothesis concerning the influence of arousal on memory was supported. These data show that caffeine is a useful method for manipulating arousal in the laboratory without influencing feelings of pleasantness or learning and memory performance. PMID- 10458051 TI - The Overvalued Ideas Scale: development, reliability and validity in obsessive compulsive disorder. AB - The presence of overvalued ideas in obsessive-compulsive disorder (OCD) has been theoretically linked to poorer treatment outcome [Kozak, M. J. & Foa, E. B. (1994). Obsessions, overvalued ideas and delusions in obsessive-compulsive disorder. Behaviour Research and Therapy, 32, 343-353]. To date, no measures have been developed which quantitatively assess levels of overvalued ideas in obsessive-compulsives. The present studies examined the psychometric properties of a scale developed to measure this form of psychopathology, the Overvalued Ideas Scale (OVIS). In study 1, 102 patients diagnosed with OCD were administered a battery of instruments including the OVIS at baseline and two weeks later, prior to initiating treatment. Results indicate that the OVIS has adequate internal consistency reliability (coefficient alpha = 0.88 at baseline), test retest reliability (r = 0.86) and interrater reliability (r = 0.88). Moderate to high levels of convergent validity was found with measures of obsessive compulsive symptoms, a single item assessment of overvalued ideas and psychotic symptoms. Medium levels of discriminant validity with measures of anxiety and depression was obtained in this study. Individuals determined to have high OVI showed greater stability of this pathology than those with lower OVI, suggesting that overvalued ideas are stable for extreme scorers. In study 2 a total of 40 patients participated who were diagnosed with OCD. The same battery of instruments was administered as in study 1, as well as the Beck Depression Inventory and Beck Anxiety Inventories. Results were similar to that obtained in study 1, including a relative lack of discriminant validity with self-report measures of depression and anxiety. It is suggested that further research with the OVIS may show predictive value in treatment outcome studies of OCD. PMID- 10458052 TI - Factor structure of the childhood anxiety sensitivity index. AB - We developed various factor models of the Childhood Anxiety Sensitivity Index [Silverman, W. K., Fleisig, W., Rabian, B. & Peterson, R. A. (1991). Childhood anxiety sensitivity index. Journal of Clinical Child Psychology, 20, 162-168] and tested the goodness of fit of the models in an independent sample. Of primary interest was to examine the question that characterized the factor analytic studies conducted on the adult version of the anxiety sensitivity index, i.e. the ASI [Reiss, S., Peterson, R. A., Gursky, D. M. & McNally, R. J. (1986). Anxiety sensitivity, anxiety frequency and the prediction of fearfulness. Behaviour Research and Therapy, 24, 1-8]: is anxiety sensitivity in children a unidimensional construct, an orthogonal multidimensional construct, or a hierarchical construct? Two independent samples (a clinic sample and a nonclinical sample) were used for development and replication of the factor models. The clinic sample consisted of 258 children (105 girls and 153 boys) who presented to a child anxiety disorders specialty clinic. The unselected, nonclinic sample consisted of 249 children (122 girls and 127 boys) enrolled in an elementary school. The results provided strong empirical support for a hierarchical multidimensional model with either three or four first-order factors. The two factors that emerged that appeared to be robust were Physical Concerns and Mental Incapacitation Concerns. What remains unresolved is whether Control of anxiety symptoms and Social Concerns are to be differentiated (as in the hierarchical model with four first-order factors) or not (as in the hierarchical model with three first-order factors). In addition to discussing this issue, the convergence of the present study's findings with past findings obtained with the ASI is discussed. PMID- 10458053 TI - 90 years of Buerger's disease--what has changed? AB - Thromboangiitis obliterans or Winiwarter-Buerger's disease is a primary systemic vasculitis of an unknown etiology, which affects medium-sized arteries and veins mainly in the lower and upper extremities, causing multiple segmental arterial occlusions especially in young male smokers. The aim of our study is to compare the knowledge on the etiology, epidemiology, clinical presentation, diagnostic and therapeutic possibilities in the time of Leo Buerger (90 years ago) and now. Between 1994 and 1998, 26 patients (19 men and 7 women) were investigated with clinical suspicion for Winiwarter-Buerger's disease. Laboratory and arteriographic investigation revealed typical signs for this disease in 22 of them. To the most common clinical signs or symptoms belong smoking and the onset of the disease before the age of 50 years (in 95.5%), intermittent claudication (in 72.7%), rest pain and ischaemic ulcers or gangrenes in the fingers (in 68.2%). In slightly more than half of the patients migrating superficial thrombophlebitis was present and similarly in one half of the patients Raynaud's phenomenon was found. In conclusion--What has changed from the times of Leo Buerger? 1. Prevalence of TAO increased in women. 2. Older patients (more than 40 years old) are being diagnosed. 3. Upperextremity involvement is more frequently present. 4. Diagnosis of TAO is being more proper, especially due to up-to-date diagnostic methods, like digital subtraction angiography. 5. The treatment is more effective, amputation number is decreased. And what has not changed? Similarly like Leo Buerger we do not known the precise etiology of the disease. Ceasation of smoking has still the most important therapeutic procedure. The clinical course of the disease is individual and in spite of the treatment is the clinical course unpredictable. (Tab. 5, Ref. 47.) PMID- 10458054 TI - [Immunostimulating activity of vaccines in the treatment of recurrent urinary tract infections. I]. AB - The authors inform about the immunomodulatory properties of the vaccine URVAKOL aimed for the treatment of recidiving urinary infections. The results of immunostimulatory activity of the preparation and its effects on cellular and humoral immunity in mice following intraperitoneal administration of the vaccine are presented. The vaccine markedly increases cytotoxic activity of adhering peritoneal cells and has protective effects in model infection induced by intracellular pathogen Francisella tularensis (strain 15 L). (Tab. 6, Fig. 6, Ref. 9.) PMID- 10458055 TI - [Antibacterial peptides isolated from the urine of healthy adults]. AB - Gel chromatography on Sephadex G-25, DEAE Sephadex A-25 and electrophoresis were used for isolation of two peptides from urine of healthy adults. Isolated peptides exhibited a higher antibacterial activity than antibiotics penicillin and tetracycline. The inhibitory zone of these peptides is similar to that of ampicillin. Chemical analysis of the isolated peptides revealed that one is composed of 5 and the other of 7 amino acids. In both peptides was the concentration of glycine the highest from all amino acids. the highest. (Tab. 1, Fig. 4, Ref. 22.) PMID- 10458056 TI - [Immunogenicity of recombinant interferon alpha-2 in therapy]. AB - Recombinant human (r)IFN-alpha 2 is successfully used in treatment of haematopoietic malignancies, lymphomas and viral hepatitis B and C as well. One of undesired side-effects of rIFN-alpha 2 is its immunogenicity which could decrease therapeutic potential of the drug. Formation of antibodies against rIFN alpha 2 in man represents a complex process, in which many mutually interacting variables are involved. Factors influencing the humoral response against the recombinant homologue of human IFN-alpha 2 have not been unambiguously determined yet. In general, two categories of these factors--exogenous and endogenous (physiological)--are considered. The exogenous factors determine structural differences of the recombinant protein from the native IFN-alpha 2. Their influence on the formation of therapy-induced antibodies could be limited by the selection of suitable rIFN-alpha 2 subvariant, suitable storage of the preparation and by treatment regimen. On the other side, current knowledge of the endogenous factors, which function at the level of patient's organism, do not allow to propose efficient ways of their elimination. (Tab. 2, Ref. 28.) PMID- 10458057 TI - [Neurodegeneration and nitric oxide]. AB - NO appears to play a significant role in the physiological processes of many of the body's systems. This review examines the present molecular, physiological and pathological knowledge related to NO and its clinical implications. The role of NO in pathophysiology of diseases is not yet fully elucidated. It is still unclear whether alterations in NO production, release and degradation are primary events in pathological processes. The presented work deals with neurodestructive and neuroprotective effects of NO. The hypothesis suggests that under physiological conditions NO acts as a neuronal messenger molecule. In pathological conditions, with excessive NO release, NO may function as a cytotoxic molecule being involved in several neurodegenerative processes. The most important issue associated with NO research will be to elucidate the cellular and molecular mechanisms of NO action. (Fig. 2, Ref. 32.) PMID- 10458058 TI - [Endothelial dysfunction and lipid profiles: analysis of the EDO Study]. AB - On the basis of experimental as well as clinical observations, endothelial dysfunction is (defined as impaired or absent endothelium-dependent relaxation) considered to be an important factor in the atherogenesis. Serum lipids abnormalities have been accepted as an epidemiological risk factor of atherosclerosis. In vitro, experimental as well as epidemiological studies revealed the fact, that lipoprotein oxidation plays an important role in atherogenesis. Recently invented non-invasive methods to test and measure the endothelial function in vivo opened the opportunity to study the influence of different serum lipids on the endothelial function directly. Therefore, we decided to employ this non-invasive method for studying the endothelial function and observe the influence of various levels of plasma lipids and lipoprotein oxidation on the endothelial function of arteries in middle-aged men, since they are the most endangered part of population. In our study we used a method of measuring the diameter of a. radialis by high-resolution ultrasound (Sonoline 450, Siemens, Japan) and further mathematical and statistical analysis of functional as well as relative vasodilation reserve followed these measurements. Blood samples were taken within 24 hours of ultrasonography to study serum lipids (total cholesterol, HDL, triglycerides) and parameters of oxidation/antioxidation (superoxid dismutase--SOD, malondialdehyde--MDA). Sixty men, 25-45 years old, from an area of basically the same level of pollution were examined. We found a negative correlation between FVDR and TCH (p = 0.01), FVDR and Tg (p = 0.002) and FVDR a TCH/HDL (p = 0.015). Positive correlation exists (p < 0.001) between TCH, Tg levels and TCH/HDL ratio and MDA level in all cases. Analysing further data from the EDO Study, we can conclude, that increased plasma lipids are more likely to be oxidized, which, in turn, is the probable reason of endothelium-dependent vasodilation impairment. PMID- 10458060 TI - [Ocular manifestations of toxocariasis]. AB - The paper reports about the ocular symptomatology of toxocariasis that represents a severe parasitic disease especially in children. Recently, the incidence of this disease is increasing. Diagnostic process has improved by means of newly developed laboratory methods. Ocular findings on retina are in toxocariasis identified very late. Despite many antihelmintics, steroids and surgical treatment, a poor treatment success has been achieved, and the sight remains often permanently severely affected. Because of the risk of blindness the most efficient arrangement is prophylaxy from the side of parents, teachers, veterinarions and the society as a whole. (Fig. 3, Ref. 6.) PMID- 10458059 TI - [Pharmacologic cytoprotection of the central nervous system]. AB - Ischaemia of the central nervous system is considered to be a phenomenon, which manifests after the ischaemic episode and has limited therapeutic possibilities. Clinical situations in which the oxygen supply may be reduced include the following: stroke and transient ischaemic attacks, subarachnoid haemorrhage before, during and after carotid artery surgery, prolonged cardiopulmonary bypass during heart surgery, head injuries, cardiac arrest: (during arrest and recirculation), aortic surgery, spinal cord ischaemia, severe hypoxia/hypoxaemia, severe hypoperfusion/shock conditions. Although these clinical conditions are markedly different, they share the major similarities of cell death and its pathophysiology and pathogenesis, to which lies the therapy of CNS ischaemia is aimed. New cytoprotective drugs for neuronal damage are lazaroids. The 21 aminosteroids are known as inhibitors of lipid membrane peroxidation which preserve post-injural Ca2+ homeostasis (after total and focal ischaemia). During the last few years evidence has been accumulated about CCAs brain protection against cerebral ischaemia by a virtue of direct neuronal action. The excitatory amino acid antagonists, barbiturates, corticosteroids, antiepileptic drugs and antioxidants complete the cytoprotective treatment possibilities in situations of reduced oxygen supply for CNS. (Ref. 17.) PMID- 10458061 TI - [Prophylactic effects of ceftriaxone in patients after endoscopic papillosphincterotomy]. AB - In thirty patients after EPS and gall extraction for cultivation ceftriaxon in preventive dose 1 g was administered. This group of patients was compared with a group of 30 patients after EPS without preventive administration of antibiotic from clinical and biochemical point of view. Most frequently occurring bacteria in the gall of patients after EPS were Pseudomonas aeruginosa and E. coli. All of the detected bacteria were sufficiently sensitive to ceftriaxon. Preventive effect of ceftriaxon was manifested in statistically significant fall of hyperbilirubinemia and hyperamylasemia 24 hours after EPS (p > 0.05). (Tab. 5, Ref. 8.) PMID- 10458062 TI - [Carcinoids--personal experience with surgical therapy]. AB - The authors retrospectively studied a group of 8 patients in the period of 7 years (1990-1997). Most frequent surgical procedure performed was appendectomy (4 cases), right-sided hemicolectomy (2 cases), gastric resection (1 case) and small intestine resection (1 case). The most frequent side of carcinoid localization was appendix followed by small intestine. In 3 cases the tumours were macroscopically detectable, the remaining only histologically. Patients were without clinical manifestations of carcinoid syndrome. (Tab. 1, Ref. 9.) PMID- 10458063 TI - Cyclic patterns of variations in incidence rates of syphilis and gonorrhoea in Slovakia. PMID- 10458064 TI - Botulism in Canada--summary for 1997. PMID- 10458065 TI - Impact of vaccines universally recommended for children--United States, 1990 1999. PMID- 10458066 TI - Hepatitis C--prevention and control: a public health consensus. Ottawa, Canada, October 14-16, 1998. PMID- 10458067 TI - [Vitrectomy and autologous thrombocyte concentrate in the treatment of idiopathic macular holes]. AB - OBJECTIVE: To evaluate the results of surgery of idiopathic macular holes (IMH) using or not using autologous thrombocyte concentrates (ATC). METHOD: Fifty eyes with IMH, stage 2-4, were divided into two groups. Group 1 comprised 27 eyes with an average persistence of complaints of 12 months where pars plana vitrectomy (PPV) and tamponade with 16% C3F8 were combined with the use of ATC. Group 2 comprised 23 eyes with a mean duration of complaints of 14 months which were treated by PPV and 16% C3F8 without ATC. In all IMH stage 2 and 3 the posterior vitreous membrane was detached from the retina. The epimacular membrane was removed, if present. The mean follow-up period in group 1 was 17.2 months and in group 2 12.3 months. RESULTS: In group 1 anatomical success was achieved in 21 eyes (78%), visual acuity improved by two lines and more in 16 eyes (59%). In group 2 anatomical success was recorded in 15 eyes (69%) (p = 0.326) and visual acuity improved in 10 eyes (44%) (p = 0.265). Nine eyes were reoperated with ATC. Reoperations improved the anatomical success in the whole series to 90% and the functional success to 62%. The visual acuity improved in 70% operated eyes within 12 months from the onset of complaints and in 47% eyes with a longer case-history (p = 0.1182). Complications included retinal tears in 5 eyes (10%), detachment of the retina in one eye (2%), minor facete-like defects in the pigmented epithelium in 22 eyes (44%), disciform maculopathy in 5 eyes (10%) and atrophy of the optic disc in 3 eyes (6%). CONCLUSIONS: PPV is beneficial in IMH in the majority of patients. ATC can improve anatomical and functional results. Functional success is more likely after surgery in the early stage of the disease. The results of surgery are jeopardized by various complications. It is important to inform patients on the limitations and risk of surgery. PMID- 10458069 TI - [Transmission electron microscopy of the surface of an intraocular lens removed from the eye]. AB - The authors examined by transmission electron microscopy the surface of an intraocular posterior chamber lens explanted from the eye of a female patient on account of secondary glaucoma. They found a fibrous reaction in particular at the site of contact of the supporting haptic with the sulcus of the ciliary body which was the cause of failure of conservative treatment. PMID- 10458068 TI - [Vitreoretinal surgery in diabetic patients in a dialysis program]. AB - Pars plana vitrectomy in patients with severe renal and retinal affections was considered a relative contraindication for operation. Despite relative failure (deterioration of visual acuity) it is possible in these patients, and in half of them it improved their quality of life. The risk of peroperative and postoperative damage can be reduced by suitable selection of anticoagulants. The favourable effect of tamponade with silicon oil on the postoperative course must be tested in a controlled trial. PMID- 10458070 TI - [Pachymetry in patients with glaucoma]. AB - Central corneal thickness (CCT) was measured in 328 eyes of 167 patients (50 men and 117 women, an average age was 62 years). CCT was determinated by ultrasonic pachymeter NIDEC, design UP-1000, at Eye Department of 3rd Medical Faculty of Prague. RESULTS: The mean CCT at glaucoma patients was 561 microns, in margins 490-650 microns. CCT for right eyes were 0.567 +/- 0.035 mm, for left eyes 0.569 +/- 0.037 mm. There were no influence by previous operations (cataract, trabeculectomy and combined operations). NTG had the lowest CCT values 551 microns, POAG had 569 microns and OHT the greates value, CCT is 588 microns. CCT in patients with NTG was significantly lower (p = 0.047) than in POAG. The higher difference is between NTG a OHT (p = 0.015). We had no statistical difference in CCT between hypermetropic, emetropic and myopic eyes. The corneal thickness is decreasing during ageing. CCT is another clinically helpful factor for evaluation our glaucoma patients. CCT together with other risk factors can help us in establishing a target pressure. Patients after refractive surgery (PRK, LASIK) should be more carefully evaluated not to miss early stages of glaucoma. PMID- 10458071 TI - [Photorefractive keratectomy in high myopic anisometropia in children]. AB - The authors evaluate the results of photorefractive keratectomy in children with high myopic anisometropia proved by intolerance of a contact lens and intolerance of full correction by glasses. The objective was to maintain or improve the already achieved binocular vision without or with feasible correction by glasses. The group is formed by 13 children, mean age 11.5 years at the time of surgery (range 7-15 years). The mean preoperative value of myopia was -8.9 dpt (SE), within the range of -7.0 to -11.75 dpt (SE) on the operated eye. The preprequisite condition for surgery was confirmed intolerance of a contact lens, intolerance of complete correction by glasses and previous systematic pleoptic treatment with training of binocular functions. The operation was always implemented during hospitalization. The follow-up after surgery in all children of the group is more than 2 years. The mean value of the refraction defect two years after surgery is -1.12 dpt (SE). The mean non-corrected visual acuity improved from 0.021 before surgery to 0.48 two years after surgery and the mean best corrected visual acuity improved from 0.51 before surgery to 0.61 two years after surgery. The authors compare results of binocular functions before and after operation. Two years after surgery all children had superposition and a normal Bagolini test, 12 children had fusion I, 9 children fusion II, and 6 children fusion III and stereopsis. Based on the results of the trial PRK appears to be an effective and safe method for correction of high myopic anisometropia in intolerance of contact lenses or complete correction by glasses. This operation thus makes it possible to preserve further the degree of binocular vision practised in advance or to improve it. This is the first group, though small, of thus treated children published in our professional literature. PMID- 10458072 TI - [Assessment of adhesion of the corneal lamella to the stroma in relation to the duration of the LASIK postoperative period]. AB - Adhesion of lamella created by two different microkeratomes was assessed during the healing period of 250 days. Oscillating microkeratome Flapmaker, Refractive Technologies and Draeger rotating microkeratome, Storz were used for this study. Increase of lamella adhesion during the healing period was found. The increase is little faster during the first weeks. The difference in adhesion between eyes operated by different microkeratome was not significant. PMID- 10458073 TI - [LASIK--comparison of clinical results in operations performed with oscillating and rotating microkeratomes]. AB - Clinical results of LASIK with two different microkeratomes were evaluated. Oscillating microkeratome Flapmaker, Refractive Technologies and rotating Draeger microkeratome, Stroz were used for this study. Operations were performed on two group of high myopic patients (-11.09 D sf.eq., -12.03 D sf.eq. mean preop. refraction). Faster visual recovery and less complications were found with Flapmaker. LASIK created clinically acceptable results in both groups of patients. PMID- 10458074 TI - [Contrast sensitivity in type I diabetics without symptoms of diabetic retinopathy]. AB - The objective of the work was to assess whether in patients with IDDM without signs of diabetic retinopathy changes in the sensitivity function to contrast (CS) develop as compared with a group of healthy subjects matched for age but also as a result of the persistence of IDDM or the level of its long-term compensation investigated by means of glycated haemoglobin (HbA1c). CS was examined by the static method using a VCTS 6500 table from a distance of 208 and 420 cm which covered the spatial frequencies from 1.15 to 27.25 c/st. Moreover the authors assessed the best correctable central visual acuity on Snellen's optotypes, the ophthalmoscopic finding on the fundus with a colour photograph, fluorescein angiography and the HbA1c serum level. A group of 49 patients with IDDM was divided into two age groups: A(under 35 years), B (35 years and above). A marked decline of the CS function occurs in all spatial frequencies in both age groups. On comparison of the persistence of IDDM in group B no significant difference was found in CS, while in group A statistical significance was revealed in spatial frequencies 4 and 27.25 c/st. On comparison of the CS function in diabetics with a normal and pathological HbA1c level it was found that long-term hyperglycaemia has not a marked effect on visual functions. CONCLUSION: The decline of CS in patients with IDDM without signs of diabetic retinopathy is influenced in particular by the patient's age, in junior patients also by the persistence of IDDM. Examination of the CS is a simple non-invasive method suitable for screening of early affections of the eyes in IDDM. PMID- 10458075 TI - [Iris cysts]. PMID- 10458076 TI - [Human cytomegalovirus (HCMV) HCMV retinitis]. PMID- 10458077 TI - Tuberculosis in a dentist. PMID- 10458078 TI - Responding to MRSA infection on surgical wards. PMID- 10458079 TI - Urinary porphyrin excretion measurements in healthy neonates. AB - The objective of this study was to establish normal reference ranges for porphyrins in healthy neonates. There is little information about urinary porphyrin excretion in this age group. This knowledge may provide an early diagnostic tool for detecting subtle alterations or latent forms in disorders of heme biosynthesis. Fifty healthy neonates were selected from the Department of Obstetrics. Total urinary porphyrins in random specimens were analysed by a spectrofluorometry method. The measurement of porphyrin fractions was made by fluorometric high-performance liquid chromatography (HPLC). The results were adjusted to urinary creatinine excretion to correct any imprecision and interindividual variation in body mass. The urinary total porphyrin had a median value of 331.50 (nmol/L). A statistically significant relationship between total porphyrin (nmol/L) and creatinine (mmol/L) was found (p < 0.01). The porphyrin/creatinine ratio showed a median value of 56.30 nmol/mmol creatinine. The study of individual porphyrins revealed that coproporphyrin and uroporphyrin were the major porphyrins excreted in neonates (coproporphyrin represents 81.98% and uroporphyrin 16.64% of total porphyrin); in both cases, isomer I was predominant with median values of 22.36 and 6.25 nmol/mmol creatinine, respectively. No significant relationships were found between porphyrin excretion and sex, gestational age, weight, or height. Our data provide the reference limits for porphyrins in neonates as a diagnostic guideline for evaluation of subtle alterations in heme biosynthesis. PMID- 10458080 TI - An immunohistochemical study of E-cadherin expression in human squamous cell carcinoma of the skin: relationship between decreased expression of E-cadherin in the primary lesion and regional lymph node metastasis. AB - E-cadherin is a Ca(2+)-dependent, intercellular adhesion molecule that is specifically expressed in epithelial tissues and is essential for maintaining intercellular connections. It has been reported that E-cadherin expression of tumor cells is often decreased in some types of metastasizing carcinomas as compared with those without metastasis. We immunohistochemically examined the expression of E-cadherin with anti-E-cadherin monoclonal antibody and compared primary lesions of human squamous cell carcinoma of the skin (SCC) with regional lymph node metastasis to those without regional lymph node metastasis. Tumor samples from fifty-five cases of SCC (32 cases of SCC without metastasis and 23 cases with metastasis) were formalin-fixed, paraffin-embedded, and examined. E cadherin was reduced or absent in 39 (70.9%) out of 55 cases of SCC, and in 21 (91.3%) of 23 cases with regional lymph node metastasis. Our results suggest that the decreased expression of E-cadherin in the primary lesion is correlated with regional lymph node metastasis in SCC and that it is more frequently correlated with well-differentiated than with poorly differentiated SCC. E-cadherin may be useful as a marker for metastatic potential in well-differentiated SCC. PMID- 10458081 TI - Pulmonary function changes in patients with psoriasis on methotrexate therapy. AB - Methotrexate is known to induce pulmonary fibrosis. The present study was undertaken to assess pulmonary toxicity, if any, in psoriasis patients on methotrexate therapy and correlate the dose and duration of the treatment with any changes in pulmonary function. Ten patients who had taken methotrexate for one year and ten patients receiving methotrexate were included in the present study. Detailed lung function studies including arterial blood gas analysis were carried out. In both groups, the only pulmonary function abnormalities detected were FEF200-1200, FEF25-75%, residual volume (RV), and RV/TLC%, which showed a decline after six months of treatment. However, this was not significant. No changes were detected in the arterial blood gas values following methotrexate therapy. Therefore, the present study did not find any significant deterioration of lung functions in psoriasis patients on methotrexate therapy. PMID- 10458082 TI - Elevated B-lymphocyte levels in lesional tissue of non-arthritic psoriasis. AB - Psoriasis is a chronic inflammatory skin disorder characterized clinically by maculopapular skin lesions and on the cellular level by increased T-lymphocyte activation in the peripheral blood and migration of activated T-lymphocyte into the lesions. The lymphocyte subpopulations in peripheral blood from 21 Kuwaiti patients showed elevated levels of the T-lymphocyte activation marker CD25, as well as increased expression of HLA-DR compared with a group of age and sex matched controls, confirming published findings on psoriasis. In addition, there was a tendency towards a significant increase in the CD4+/CD45RO+ (memory cell) population that was also consistent with peripheral T-lymphocyte activation. Immunohistological studies showed a heavy infiltrate of all cell types into the lesional tissue including, as expected, activated T-lymphocytes. An unexpected finding was significantly higher levels of B-lymphocytes infiltrating the psoriatic lesions; they numerically exceeded the T-lymphocyte infiltrate. This has previously been reported only in cases of psoriasis with concurrent arthritis. None of the subjects had arthritis, suggesting an immunopathological variant of psoriasis possibly specific to this population group. PMID- 10458083 TI - Detection of human herpesvirus 8 in Korean Kaposi's sarcoma cases by polymerase chain reaction and in situ polymerase chain reaction. AB - Several infectious agents, including herpesvirus-like particles, had been suggested as possible candidates for the development of Kaposi's sarcoma (KS), and a new herpesvirus, human herpesvirus 8 (HHV-8), was recently identified in the vast majority of KS lesions, irrespective of their association with human immunodeficiency virus (HIV) infection. However, the etiologic role of HHV-8 in KS remains controversial. We undertook this study to screen for and localize the presence of HHV-8 in KS in Korea. A total of 46 paraffin-embedded specimens were studied, including KS, hemangioproliferative disorders, and 10 non-KS lesions from HIV-positive patients. We performed nested polymerase chain reaction (PCR) and in situ PCR with HHV-8 specific primers. HHV-8 DNA sequences were detected in 8 of 11 KS specimens. All specimens of hemangioproliferative disorders, non-KS lesions from HIV-positive patients, and other skin samples were negative for HHV 8. When sequencing PCR products, the sequences were almost identical with the prototypic sequence for HHV-8. In PCR-positive tissues, in situ PCR staining of HHV-8 localized to nuclei of endothelial cells and perivascular spindle-shaped tumor cells. The results of this study suggest that HHV-8 is not widespread and has a certain causative role in the development of KS. Further studies, including serological and animal studies, will be helpful to appreciate an epidermiological link and pathogenetic mechanism between HHV-8 and KS. PMID- 10458084 TI - Chronic pruritic papular dermatitis in adult men: a variant of prurigo. AB - Chronic recurrent pruritic papular eruptions in which a specific diagnosis cannot be established becomes a baffling experience to the dermatologist. We have met adult male patients with chronic recurrent pruritic papular eruptions, but their clinicopathological features are not described in English language textbooks. Our purpose was to study the clinical and histological features of this entity and review the various existing taxonomy. We conducted a study of 20 patients over a six year period by taking histories, performing skin biopsies, screening patch tests, and doing immunofluoresence studies. The eruptions occurred exclusively in male adults and had a predilection for the trunk and proximal extremities. The lesions were characterized by severely pruritic, nonfollicular, monomorphic, erythematous urticarial papules. There was no evidence of atopic diathesis or history of insect bite. Most patients had normal levels of serum eosinophils and IgE. The predominant histopathologic finding was a presence of perivascular infiltration of mononuclear cells with eosinophils. The patients followed a chronic course of at least six months with waxing and waning; systemic corticosteroids were the only effective treatment. Finally, all other pruritic erythematous papular dermatoses were ruled out. These cases comprise a distinct entity that has previously been mentioned in a few reports. Clear definition of this entity with an appropriate designation is in order to avoid confusion among dermatologists, and we propose the disease name "chronic papular dermatitis in adult men" as a variant of prurigo. PMID- 10458085 TI - Epstein-Barr virus-associated lymphoproliferative skin lesion with recurrent necrotic papulovesicles of the face. AB - A 24-year-old man showed recurrent necrotic papulovesicles of the face. The cutaneous lesions started with erythema, followed by vesicles, necrosis, and crusting, and leaving depressed scars. Light avoidance did not prevent the development of the lesions. Histologic examination revealed lymphoproliferative lesions confined to the skin. Latent Epstein-Barr virus (EBV) infection was detected in the lymphoid cells from the skin lesions by in situ hybridization. A moderate to high dosage of corticosteroids suppressed the development of new skin lesions, but relapses occurred when these systemic corticosteroids were tapered. PMID- 10458086 TI - Erythema elevatum diutinum complicated by rheumatoid arthritis. AB - A 53-year-old female developed erythema elevatum diutinum (EED) twelve years after the onset of rheumatoid arthritis. The arthritis had been well controlled for the last several years. Annular purpuric macules were characteristically complicated by common nodular and plaque lesions. Both leukocytoclastic vasculitis and fibrosis were observed in the macular lesions, indicating that the lesions were a manifestation of an early phase of EED. Both types of skin lesions disappeared with treatment with dapsone. They have not relapsed for two years after stopping the dapsone. The leukocytoclastic vasculitis was thought to have developed independently of the rheumatoid arthritis. She had noticed sicca symptoms two years before the appearance of EED, but she did not satisfy the diagnostic criteria for Sjogren's syndrome. PMID- 10458087 TI - Low-dose intravenous immunoglobulin therapy for intractable dermatomyositis skin lesions. AB - We report a 45-year-old female patient with dermatomyositis whose corticosteroid resistant cutaneous manifestations improved with low-dose (0.1 g/kg body weight/day for 5 days weekly for 2 weeks) intravenous immunoglobulin (IVIG) therapy. PMID- 10458088 TI - Methotrexate nodulosis. AB - Methotrexate (MTX) nodulosis in patients with rheumatoid arthritis treated with MTX has become a well recognized phenomenon. It has not been described in patients receiving MTX for treatment of other diseases, e.g., dermatological or malignant diseases. Recently, MTX nodulosis was described in a patient with psoriasis and arthritis. The pathophysiology and treatment of MTX nodulosis are yet unsettled. We experienced a case with dermatomyositis who developed multiple subcutaneous nodules after treatment with MTX. In our patient, numerous, small, symmetrically distributed, grouped subcutaneous nodules were seen on a V-shaped area of the anterior chest, both axillary areas, and the medial sides of both upper arms and thighs. We histopathologically examined a nodule and started treatment with hydroxychloroquine after discontinuation of MTX. The histopathology revealed septal panniculitis, and the nodules have been under regression. MTX nodulosis may be a drug-specific phenomenon. Discontinuation of MTX and replacement with hydroxychloroquine are recommended in severe cases. Awareness of this entity is important for diagnosing and treating cases of multiple nodules developing after the administration of MTX, which is widely used for many dermatological diseases. PMID- 10458089 TI - A case of pre-Sezary syndrome preceded by hand lesions. AB - Pre-Sezary syndrome is an erythroderma with a chronic course, clinical findings of Sezary syndrome, lymphocytic subepidermal band infiltration at times, and repeated cycles of circulating Sezary cells of less than 1,000 cells/mm3. Duration of the pre-existing skin diseases preceding pre-Sezary erythroderma varies from a few weeks to 20 years. Before the erythroderma develops, these patients are diagnosed with contact dermatitis, neurodermatitis, chronic dermatitis, atopic dermatitis, or asteatotic eczema. Hand lesion also precedes the pre-Sezary erythroderma. This condition has been controlled by three cycles of chemotherapy consisting of vincristine, cytoxan, doxorubicin, and prednisolone. We describe a case of pre-Sezary syndrome preceded by hand lesion and treated with chemotherapy. PMID- 10458090 TI - Giant cell (temporal) arteritis involving both external and internal carotid arteries. AB - A 76-year-old woman with giant cell (temporal) arteritis was described; she presented with a one year history of headache and tinnitus. Histopathological findings from a superficial temporal artery showed arteritis with granulomatous changes. Bilateral carotid arteriograms demonstrated the stenoses of both internal carotid arteries as well as the narrowing of the superficial temporal arteries. Although we dermatologists rarely encounter the disease in daily clinical practice, it is of clinical importance to perform cerebral angiography in patients suspected of temporal arteritis. PMID- 10458091 TI - Lymphangioma circumscriptum masquerading as irritant contact dermatitis. PMID- 10458092 TI - Determination of bovine beta2-microglobulin and albumin in urine by a reversed phase high-performance liquid chromatography. AB - Reversed-phase high-performance liquid chromatography (HPLC) was used to analyze beta2-microglobulin and albumin in bovine urine. The urine samples were chromatographed on TSK-gel ODS-120T column with an acetonitrile gradient. Urinary beta2-m and albumin were detected at 220 nm. For the pre-treatment, there were two steps proceeding injection: dialysis of urine with distilled water overnight, followed by concentration by solid-phase extraction method using a Sep-Pak cartridge. The retention times of beta2-microglobulin and albumin were 25.35 +/- 0.85 and 32.20 +/- 0.20 minutes (n=5), respectively. The mean analytical recoveries of beta2-microglobulin and albumin added to 0.1 ml of urine samples were 94.5 and 100.5%, respectively. The within-run coefficients of variation ranged from 1.5 to 5.3% for beta2-microglobulin and from 2.3 to 7.0% for albumin. The sensitivity for quantification of each protein was 0.5 microg in 100 microl injected urine samples. Urine samples from healthy cows and from cows with different types of proteinuria were analyzed by this reversed-phase HPLC. Results revealed albumin was remarkable in the urine from a cow with glomerulonephritis, and beta2-microglobulin was, in the urine from a cow with tubular dysfunction. PMID- 10458093 TI - Canine coronavirus infections in Japan: virological and epidemiological aspects. AB - Ten strains, eight field and two reference laboratory strains, of canine coronavirus (CCV) were comparatively examined with respect to antigenic relationships and pathogenic potential in dogs. With monoclonal antibodies and hyperimmune antisera to feline coronavirus and CCV, respectively, varying degrees of antigenic diversities were found among the strains by neutralization and immunofluorescence assays, but it was felt that they belong to one serotype. Specific-pathogen-free puppies experimentally inoculated with some CCV strains manifested clinical symptoms, but there was a difference in their virulence. In order to elucidate the prevalence of CCV infections in dogs in Japan, we tested for neutralizing antibodies to CCV in 467 field dogs, and found a prevalence of 44.1%. Moreover, by using nested reverse transcriptase-polymerase chain reaction on rectal swabs of 100 diarrheic dogs recently presented in veterinary clinics, evidence of CCV in 16% of these specimens was found. The results suggested that CCV infection is more widespread than expected in dogs, and that CCV is a significant etiologic factor in canine diarrhea also in Japan. PMID- 10458094 TI - An investigation of the relationship between duct system and A cell-rich and PP cell-rich pancreatic islets in the canine pancreas. AB - The pancreata of four six-month-old dogs of the same mother, two with both the pancreatic and accessory pancreatic ducts (X-type) and two with only the accessory pancreatic duct (Y-type), were examined in this study. To clarify the relationships between the type of pancreatic duct system and the composition of pancreatic endocrine cells, the pancreata were examined immunohistochemically using antiserum against four types of pancreatic hormones (glucagon, insulin, somatostatin and pancreatic polypeptide). In all areas of the X- and Y-type duct system pancreata, B cells accounted for 52-82% of the total number of islet cells, and D cells accounted for 4-15%. In the X-type ducts system, the percentages of A and PP cells in the right and left lobes of the pancreas differed greatly. It was found that A and PP cells appear in inverse proportion to each other and that there exist A cell-rich and PP cell-rich pancreatic islets. The A cell-rich pancreatic islets appeared in the left lobes along the accessory pancreatic duct, while the PP cell-rich pancreatic islets were observed in the right lobes along the pancreatic duct. The body of the pancreas contained both A cell-rich and PP cell-rich pancreatic islets. In the Y-type duct systems, A cell-rich pancreatic islets appeared in the right lobes. These findings indicate that the composition of A and PP cells in pancreatic islets is closely related to the type of duct system. PMID- 10458095 TI - Binding characteristics of folate to high affinity folate binding protein purified from porcine serum. AB - High affinity folate binding protein (HFBP) in porcine serum was purified 2,000 fold to a specific activity of 1.4 nmol of tetrahydrofolic acid (THF) bound per mg of protein, using folic acid (FA) coupled EAH-Sepharose gel affinity chromatography. Binding activity of purified HFBP to folate was examined by ultrafiltration method linked to high-performance liquid chromatography with electrochemical detection or to liquid scintillation counting. Binding affinity of HFBP to folate was characterized by dissociation constants (Kd): 13, 17, and 31 pM for tritiated FA (3HFA), THF, and N5-methyltetrahydrofolic acid (5MF), respectively. FA, THF, and 5MF significantly inhibited binding of HFBP to 3HFA, and according to the magnitude of intensity of the binding inhibition, the order of affinity of each folate was confirmed to be FA > THF > 5MF. Binding activity was rather high and stable for THF and 5MF at pH ranging from 6.0 to 10.0. The binding activity, however, rapidly decreased at pH below 6.0 and over 10.0. No binding activity was observed pH below 3.0 and over 12. Gel filtration analysis showed that the prepared HFBP solution had specific binding activity at around 77 kDa of apparent molecular weight, which was 82 kDa by SDS-PAGE. It is considered that this specific and stable binding enables THF to distribute in porcine plasma. PMID- 10458096 TI - An epidemic of parapoxvirus infection among cattle: isolation and antibody survey. AB - A disease characterized by papules, nodules, vesicles and, rarely, pustules and ulcers on teats was seen among cattle on a farm in Chiba Prefecture, Japan. A virus was isolated by inoculation of fetal bovine lung cell cultures from a vesicle on a teat of an infected cow. The virus was subsequently passaged in fetal bovine lung and muscle cells in which it produced complete cytopathic changes. The virus was identified by physicochemical examinations and electromicroscopic observation as a parapoxvirus. A seroepidemiological survey was performed on antibody to the isolated virus by the agar gel immunodiffusion test. The isolated virus formed a precipitation line which cross reacted with other parapoxviruses isolated previously in Japan. The positive rate was more than 50% among cattle in the Kanto district. The positive rate increased with age. It was suggested that parapoxvirus infection might have already been prevalent among cattle in Japan. PMID- 10458097 TI - Testicular damage after exposure to carbendazim depends on the number of patent efferent ductules. AB - To study how long-term testicular damage depends on the patency of the efferent ductules (EFDs), rat testes and epididymides were examined after a single exposure to carbendazim (methyl 2-benzimidazole carbamate; MBC). The number of patent EFDs was determined in sections of the caput epididymides at 8, 16, 32 and 70 days post-treatment, and the testes were grouped into the following categories: those with intact EFDs, those with partially patent EFDs, or those with totally occluded EFDs. In each testis, 100 seminiferous tubules were examined for the presence of abnormalities. The mean weight of testes with partially patent EFDs was significantly higher compared with the control, whereas that of testes with totally occluded EFDs was significantly lower. Histologically, most seminiferous tubules of the testes with intact EFDs were normal. The testes with partially patent EFDs contained normal, degenerative and atrophic seminiferous tubules at various frequencies depending on the number of patent EFDs, and it was evident that as the number of patent EFDs increased, the number of normal seminiferous tubules also increased at any interval. In these testes, the number of normal seminiferous tubules increased progressively as the post-treatment interval increased, irrespective of patency of the EFDs. In the testes with totally occluded EFDs, atrophic seminiferous tubules were the most numerous. These results indicate that whether or not the testis is able to survive the long-term deleterious effects of MBC depends largely EFD patency. PMID- 10458098 TI - Immunohistochemical survey of the gut endocrine cells in the common tree shrew (Tupaia belangeri). AB - Regional distribution and relative frequency of endocrine cells in the gastrointestinal tract of the common tree shrew (Tupaia belangeri) were studied immunohistochemically. Ten types of immunoreactive endocrine cells were localized in the gastric mucosa, i.e., chromogranin-, serotonin-, gastrin-, somatostatin-, bovine pancreatic polypeptide (BPP)-, enteroglucagon-, pancreatic glucagon-, peptide tyrosine tyrosine (PYY)-, motilin-, and substance P (SP)-immunoreactive (IR) cells. In the intestine, 13 types of immunoreactive cells were observed, i.e., chromogranin-, serotonin-, somatostatin-, gastrin-, BPP-, enteroglucagon-, PYY-, secretin-, cholecystokinin (CCK)-, gastric inhibitory peptide (GIP)-, motilin-, neurotensin-, and SP-IR cells. The regional distribution and relative frequency of the cell types varied along the gastrointestinal tract. Basically, the types, distribution, and relative frequency of the gut endocrine cells were similar to those reported in other mammalian species. However, some characteristic findings were noted in the present study: (1) the considerably large number of gastrin-IR cells in the pyloric region; (2) numerous serotonin-IR cells in the stomach; (3) appreciable number of BPP-IR cells in the transitional region of the stomach; and (4) wide distribution of PYY- and motilin-IR cells in the gut. PMID- 10458099 TI - Abnormalities of extracellular matrices and transforming growth factor beta1 localization in the kidney of the hereditary nephrotic mice (ICGN strain). AB - ICR-derived strain with glomerulonephritis (ICGN) is a strain of mice with hereditary nephrotic syndrome with an unidentified cause. Based on histopathological and biochemical data, ICGN mice are considered to be a good experimental model for human idiopathic nephrotic syndrome. In the present study, we histochemically investigated the changes in localization of extracellular matrix (ECM) components and transforming growth factor beta1 (TGF-beta1). Strong immunohistochemical staining of basal membrane ECM components (collagen IV and laminin) and interstitial ECM components (type III collagen and fibronectin) were demonstrated in glomeruli and tubulointerstitum of ICGN mice as compared with those of sex and age-matched ICR mice, used as normal healthy controls. Marked type I collagen and tenascin deposition, which were not detected in the glomeruli of ICR mice, were seen in the glomeruli of ICGN mice. A remarkable increase in active-TGF-beta1 was also detected only in glomeruli of ICGN mice, but not in those of ICR mice. Furthermore, strikingly increased alpha-smooth muscle actin, a marker of activated glomerular mesangial cells, was demonstrated in the glomeruli, mainly in the mesangial cells, of ICGN mice. These findings indicated that ECM components are increased in the glomerulus and tubulointerstitum of ICGN mice, and that active-TGF-beta1 induces such increases in ECM components. The present findings may contribute to elucidation of the pathogenic mechanisms of hereditary nephrotic syndrome in ICGN mice and, in future, human idiopathic nephrotic syndrome. PMID- 10458100 TI - Diazepam-induced hyperphagia in mice is sensitive to quinpirole. AB - The present trial examined the possibility that diazepam (DZP, 1 mg/kg) induces hyperphagia by acting on the dopaminergic system. Quinpirole (QP), dopamine D-2 receptor agonist, was used for this purpose. Mice fasted for 24 hr were treated with QP 1 (QP-1) or 2 (QP-2) mg/kg 30 min prior to termination of the starvation. DZP was given to untreated mice and half of the QP-1 and QP-2 treated mice 10 min before the termination of the starvation. Food consumed during six 30 min intervals (30 min-feeding), food consumed for 3 hr (total feeding), time required to enter the room containing food by passing through a maze with four multiple routes (time to banquet), latent period to commencement of eating food after entering the banquet room (latent period), and feeding frequency for the 30 min intervals as well as for 3 hr were measured. DZP stimulated feeding, shortened the latent period without affecting the time to banquet and increased the feeding frequency. The hyperphagic effect was restricted to the first 30 min interval only. Both QP-1 and QP-2 first reduced, then progressively stimulated, and finally reduced feeding without modifying total feeding, thus making a bell shaped profile. They also prolonged both the time to banquet and the latent period, and reduced the feeding frequency of the first 30 min interval but not that for 3 hr. Both QP-1 and QP-2 canceled all the effects of DZP. These results imply that dopamine D2 receptor is involved in the induction of hyperphagia by DZP. PMID- 10458101 TI - Induction of parturition in bitches with minimal side effects by two injections of a low dose of fenprostalene, a prostaglandin F2alpha analogue, and pretreatment with prifinium bromide. AB - An experiment using 16 Beagle bitches (aged 11 months to 6 years and 2 months) in their 56th to 58th day of pregnancy was carried out to investigate the effects of two injections of a low dose of fenprostalene, a long-acting prostaglandin F2alpha analogue, and pretreatment with prifinium bromide, a parasympathetic nerve blocking agent, on the induction of parturition and severity of side effects. The bitches were divided into three treatment groups: one injection of 5 microg/kg of fenprostalene (group I, n=5); one injection of 7.5 mg/head of prifinium bromide followed by one injection of 5 microg/kg of fenprostalene at 5 min after prifinium bromide injection (group II, n=6); and one injection of 7.5 mg/head of prifinium bromide followed by two injections of 2.5 microg/kg of fenprostalene, one injection at 5 min after prifinium bromide injection and the next at 1 hr after the fenprostalene first injection (group III, n=5). Following the injection of fenprostalene, side effects such as salivation, vomiting, colic symptoms, and watery diarrhea occurred most frequently (80-100% of cases) in group I bitches. Apart from colic symptoms, no side effects were observed in group III bitches. Group III bitches also showed the smallest increase in plasma cortisol concentration. No significant difference in the time to initiation of parturition was found between the three groups. The one-week survival rate of newborn puppies was highest in group III. The results showed that pretreatment with prifinium bromide and two injections of 2.5 microg/kg of fenprostalene can alleviate side effects following fenprostalene administration and have no adverse effect on the survival of newborn puppies, indicating that this method is a reliable and safe way of inducing parturition in bitches. PMID- 10458102 TI - Identification and sequence analysis of the Marek's disease virus serotype 2 homologous genes of the herpes simplex virus type 1 UL25, UL26 and UL26.5 genes. AB - We identified and determined the nucleotide sequence of Marek's disease virus serotype 2 (MDV2) UL25, UL26 and UL26.5 homologous genes of herpes simplex virus type 1 (HSV-1). The UL25, UL26 and UL26.5 genes of HSV-1 encode virion proteins (UL25 and UL26.5) and serine protease (UL26). The deduced amino acid sequences of the three proteins show a high degree of homology to counterparts of HSV-1. By northern blot analyses we found that four transcripts whose sizes are 4.9, 3.9, 2.0 and 1.3 kb are transcribed from the domains of MDV2 genome containing the three genes. This is the first report dealing with UL25, UL26 and UL26.5 homologues of HSV-1 in MDV serotypes. PMID- 10458103 TI - The influence of chronic sympathectomy on cutaneous blood flow in the rat tail. AB - Tail blood flow (TBF) in the rat markedly increases during sympathetic withdrawal such as hyperthermia or lumbar sympathetic blockade. However, a long-term alteration of TBF after chronic sympathetic denervation is not well understood. In the present study, TBF following lumbar sympathectomy (LSX) was observed to ascertain whether subsequent changes in TBF occur in the absence of the sympathetic nervous activity in the rat tail. Assessed by recording tail and rectal temperature, the LSX immediately caused an increase in TBF. TBF was gradually decreased along with time and returned to the sham operated (SO) control level within 4 days. About a week after the surgery, a rapid increase in TBF in response to whole body heating was almost abolished in denervated animals. Neither hexamethonium (20 mg/kg, i.v.) for ganglion blockade nor intra-arterial infusion of alpha-receptor antagonist, phentolamine (10, 100 microg) produced vasodilation in LSX animals. Nitroprusside, a donor of nitric oxide, produced an increase in TBF in both LSX and SO animals. These results indicate that the tail vasculature after LSX constricts with capability to be vasodilated independent of sympathetic reinnervation. Quantification of the tail vascular mRNA expression by reverse transcriptase-polymerase chain reaction showed less endothelial nitric oxide synthetase in LSX group than that in SO group whereas endothelin-1 was not significantly different in both groups. It is suggested that functional changes in tail vascular endothelium takes at least a part in the reduction in TBF after LSX. PMID- 10458104 TI - Histopathological and immunohistochemical analysis of the endocrine and exocrine pancreas in twelve cattle with insulin-dependent diabetes mellitus (IDDM). AB - Histological and immunohistochemical studies were carried out on the pancreas of twelve cattle of insulin-dependent diabetes mellitus (IDDM). They showed clinical signs such as persistent hyperglycemia, glycosuria and decreased glucose tolerance, and some cases accompanied with or without ketonuria. Histopathologically, eight cattle were diagnosed as chronic IDDM, while others were acute IDDM. The most characteristic lesions of the pancreas in chronic IDDM showed a decrease in the size and number of pancreatic islets, interlobular and interacinar fibrosis, mild lymphocytic insulitis, and vacuolation of a few islets. Almost all cells in the atrophied islets had a small amount of ungranulated cytoplasm. Immunohistochemical examination revealed that the atrophied islet cells did not react to anti-insulin antibody, but occasionally reacted to anti-glucagon or somatostatin antibodies. A few solitary islets with mild lymphocytic infiltration, necrotic islets with occasional calcification, and atrophied islets with mild fibrosis were also observed. A few islets consisted of many islet cells with vacuolated cytoplasm including a small number of insulin positive granules. Accumulation of glycogen granules was occasionally observed in these islets. Islet fibrosis was due to the proliferation of collagen fibers reactive to both anti-collagen type I and type III antibodies. In acute IDDM, the major islets consisted of the cells with vacuolated cytoplasm indicating the degranulation of islet cells. These islets contained many islet cells with shrunken cytoplasm and karyorrhectic nuclei. Lymphocytic infiltration was frequently observed in the islets which consisted of many islet cells having karyorrhectic nuclei and vacuolated and severely degranulated cytoplasm. Immunohistochemically, islet cells with vacuolated cytoplasm had a small amount of insulin-positive granules, suggesting severe degranulation of beta-cells. An increase in acinar islet-cells and proliferation of ductal epithelial cells showing insulin-immunoreactivity were observed. Bovine IgG-immunoreactive islet cells were frequently seen in the vacuolated islets. In summary, pathological observations suggested that beta-cells were being destroyed by an inflammatory process which selectively affected the pancreatic islets. Lymphocytic insulitis and anti-bovine immunoreactive islet cells were thought to be the most significant changes in determining the etiology and pathogenesis of bovine IDDM, and suggested their role in anti-islet autoimmunity in this form of diabetes. PMID- 10458105 TI - Effect of thymus extract on immunologic reactivity of chicken vaccinated with infectious bursal disease virus. AB - The effects of crude thymus extract on the immune response and protection against challenge with virulent infectious bursal disease virus (IBDV) were studied in one-day-old chick. Oral administration of thymus extract (1 ml/kg) markedly and significantly increased the total protein, albumin, globulin, Tri-iodothyronine (T3), Thyroxine (T4) and the body weight gain in one-day-old chick. In addition, it increased the total lymphocytic count over four weeks after administration. Although vaccination also increased total protein, globulin, T4 and the total lymphocytic count but it significantly decreased the body weight gain of the chick and administration of thymus extract, before, during or after vaccination markedly improved the vaccination effectiveness with significant elevation of the globulin level and body weight gain of the chick. It also prevented the decrease in the relative weights of bursa, spleen and thyroid gland which commonly prevailed during vaccination. Chicken administered thymus extract and vaccinated with infectious bursal disease (IBD) vaccine showed 100% protection against challenge with IBDV. Meanwhile the vaccinated non-thymus treated group exhibited 80% protection against IBDV challenge. These results indicate a potentiating effect of thymus extract on the immune system in baby chick. These findings are supported by ELISA results that showed a marked increase in antibody titers in thymus treated groups. Additionally, microscopical examination of the bursa and the existent lymphoid hyperplasia in thymus treated groups but not vaccinated group support our findings. PMID- 10458106 TI - Interleukin-12, interferon-gamma and interleukin-4 gene expression in cats infected with Toxoplasma gondii. AB - Interleukin-12, Interferon-gamma and Interleukin-4 mRNA levels in cells of the spleen and mesenteric lymph nodes of cats following primary and secondary infection with Toxoplasma gondii were examined by a semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) method. Expression of Interleukin-12p40 mRNA and Interferon-gamma mRNA was observed after primary and secondary oral infection with Toxoplasma gondii. In contrast, no expression of IL 4 mRNA in the spleen and little expression in the mesenteric lymph nodes were observed after primary infection when the cats shed oocysts, however, the expression of IL-4 mRNA observed in the cats after secondary inoculation. PMID- 10458107 TI - Supplementary report on experimental autumn crocus (Colchicum autumnale L.) poisoning in cattle: morphological evidence of apoptosis. AB - Previously we reported that tissue destruction characterized by the presence of karyopyknotic, karyorrhectic and mitotically arrested cells was seen in alimentary epithelial cells and lymphocytes in the lymphoid and hemopoietic systems of cattle experimentally administered with autumn crocus (Colchicum autumnale L.). This report deals with the mechanism of acute cellular injury following experimental autumn crocus poisoning in cattle as demonstrated by the in situ DNA strand break analysis and electron microscopy. The analyses revealed that cellular injury caused by autumn crocus was closely associated with apoptosis. PMID- 10458108 TI - Detection of porcine epidemic diarrhea virus using polymerase chain reaction and comparison of the nucleocapsid protein genes among strains of the virus. AB - Two pairs of PCR primers were designed to perform nested PCR targetting of a 540 bp fragment of the nucleocapsid (N) protein gene (N gene) of porcine epidemic diarrhea virus (PEDV). The N gene of PEDV was amplified with 4 PEDV strains and 11 small intestines of PEDV-infected piglets collected from 2 farms in Kagoshima prefecture, Japan. Nucleotide sequences of the PCR products from a Korean and two Japanese strains (KKN96-1 and S1) of PEDV isolated in 1993 and 1996, respectively, were almost identical. These results suggest that the PCR is an available tool for detection of PEDV from pigs in the field, and that the two Japanese strains (KKN96-1 and S1) were genetically similar to the Korean strain. PMID- 10458109 TI - Enhancement of Borna disease virus transcription in persistently infected cells by serum starvation. AB - Transcription of Borna disease virus (BDV) in persistently infected MDCK (MDCK/BDV) cells increased in the fetal bovine serum free media as detected by Northern blot analysis. Especially, the amount of 1.9-kb RNA without cap formation at the 5' end and polyadenylation at the 3' end, increased as compared to other mRNA molecules of BDV. Growth arrest of MDCK/BDV cells observed in the condition of serum starvation might be important for increasing viral transcription. Since N-cadherin is the responsible factor for cell-to-cell contact, MDCK/BDV cells were cultured in calcium free medium which inhibits the interaction of N-cadherin. However, inhibition of cell-to-cell contact by N cadherin is not effective on up regulation of viral transcription. Our finding in this study indicates that enhancement of BDV transcription by serum starvation is a useful technique for further investigation in understanding of mechanisms of BDV transcription. PMID- 10458110 TI - Morphological observation of canine natural killer cells mediated cytotoxicity. AB - The cytotoxic effects of canine NK cells on CL-1 target cells were examined by scanning electron microscopy (SEM). NK cell mediated cytotoxicity on CL-1 target cells was detected by 51Cr release assay. SEM showed that a canine NK cell extended projections to the CL-1 target cell. Furthermore, the surface of CL-1 target cells changed a mesh-like structure. Therefore, the cytotoxic effects of canine NK cells on CL-1 target cells were morphologically demonstrated. PMID- 10458111 TI - Multicystic renal dysplasia in a Japanese black bull. AB - Multicystic renal dysplasia was found in a 6-day-old Japanese black bull. Grossly, both kidneys were markedly small (2.0 x 3.5 cm) with numerous cysts ranging from 1 to 8 mm in diameter. Histopathologically, both kidneys consisted of many irregularly enlarged cysts, immature glomeruli, small ducts and anomalous stromal connective tissues containing focal persistent mesenchyme characterized by a proliferation of stellate cells with myxomatous area. These features are compatible with those of multicystic renal dysplasia in humans and other mammals. PMID- 10458112 TI - Fluorescence expression by bovine embryos after pronuclear microinjection with the EGFP gene. AB - Fluorescence expression by bovine embryos was examined after pronuclear microinjection with an enhanced green fluorescent protein (EGFP) cDNA under control of the chicken beta-actin promoter and cytomegalovirus enhancer, as a first step in evaluating the applicability of EGFP for non-invasive selection of transgenic bovine embryos. After injection, developmental competence of the embryos was reduced, and light was emitted in 11.9% of them (37/310) under a fluorescence microscope. Although 2.9% of the injected embryos developed to the fluorescent blastocysts (9/310), a majority of the fluorescent embryos showed mosaic expression including the negative blastomeres (26/37, 70.3%). These results suggest the feasibility of EGFP for in vitro selection of transgenic bovine embryos by fluorescence microscopy. However, the impaired development and high frequency of mosaicism were observed in these injected embryos. PMID- 10458113 TI - An efficient multiplex PCR suitable for large scale typing in linkage mapping. AB - The dissection of polygenic traits is made possible with the development of microsatellite markers. Linkage study of this kind involves many markers with tens of hundreds of samples. Although typing essentially contains only two steps: PCR amplification and gel electrophoresis. Such work is still heavy when a large number of samples had to be genotyped. Multiplex PCR may reduce the work, but one has to optimize the conditions from marker to marker. Here we describe a dye compatible multiplex PCR that works under standardized condition without the need to pre-determine the combinational primer concentration and the time-consuming step to mix many samples with gel loading dye before electrophoresis. This successful protocol should greatly reduce the cost and labor for genetic study of polygenic traits. PMID- 10458114 TI - Thymic carcinoma with neuroendocrine differentiation in a calf. AB - A neuroendocrine carcinoma originating in the thymus was found in a 7-month-old, castrated male, Japanese Black calf. The neoplasm consisted largely of very primitive cells, characterized by the paucity of cytoplasmic organelles, but a few cells were immunoreactive for somatostatin or neurofilaments. The expression of both cytokeratin and neurofilament protein was a feature of neuroendocrine differentiation. This neoplasm considered to be a tumor of a thymic stem cell, with little but indubitable evidence of differentiation into somatostatin producing cells. PMID- 10458115 TI - Typing Listeria monocytogenes by random amplified polymorphic DNA (RAPD) fingerprinting. AB - Twenty epidemiologically unrelated Listeria monocytogenes strains isolated from different animals, locations and on different dates in Japan were classified into 18 types by the random amplified polymorphic DNA (RAPD) fingerprinting technique with four primers. Further, seven epidemiologically related L. monocytogenes strains isolated from raw milk and a bulk tank on a dairy farm represented the same RAPD type suggesting that they were all of the same origin. Therefore, RAPD polymerase chain reaction (PCR) analysis, which is rapid, simple and inexpensive to perform, can be used in surveys as a convenient epidemiological technique. PMID- 10458116 TI - Bioremediation of trichloroethylene and cis-1,2-dichloroethylene-contaminated groundwater by methane-utilizing bacteria. AB - Experimental studies on the bioremediation of groundwater contaminated with low concentration trichloroethylene (TCE) and cis1,2-dichloroethylene (DCE) were performed with two sets of bioreactors. Reactors No. 1 and No. 2 were operated without and with methane supplement, respectively. No inoculum was used. The concentrations of TCE and DCE in the effluent and the off gas from reactor No. 2 were much lower than those from reactor No. 1. When air and an H2O2 solution were supplied to reactor No. 2, concentrations of TCE and DCE in the effluent and the off gas were lower than the lowest detectable limit. The population of methane utilizing bacteria in reactor No. 2 was 1,000 times higher than that in groundwater or in the effluent from reactor No. 1. These methane-utilizing bacteria were apparently attributable to the treatment of TCE. PMID- 10458117 TI - Nasal sensory receptors responding to capsaicin, water and tactile stimuli in sevoflurane-anesthetized dogs. AB - Responses of nasal receptors to capsaicin and water were studied from afferent recordings of the posterior nasal nerve (PNN) in 12 anesthetized dogs. Out of 12 non-respiration-modulated nasal receptors, 7 responded only to capsaicin, 3 responded to both water and capsaicin, and 2 to neither of them. All the fibers showed a rapid adaptation to mechanical probing of the nasal mucosa. These results indicate that the presence of sensory receptors responding to capsaicin and water are involved in PNN afferents of the dog. PMID- 10458118 TI - Evolution of surgery for Crohn's disease: a century of progress. PMID- 10458119 TI - Nonoperative management of hemorrhoids: evolution of the office management of hemorrhoids. PMID- 10458120 TI - Management of iatrogenic rectourethral fistula. AB - PURPOSE: Rectourethral fistulas are uncommon, usually iatrogenic injuries that are challenging to treat. Our aim was to determine a logical approach to surgical treatment of this often debilitating problem. METHODS: Records of all patients who were diagnosed with rectourethral fistula between January 1981 and December 1995 were reviewed and 16 males were identified. All but three patients had had intervention for their prostatic malignancy performed elsewhere. All patients were interviewed by telephone to establish follow-up. The mean age of the sixteen patients was 68 years. The mean follow-up was 80 months. Adenocarcinoma of the prostate in 15 patients and recurrent transitional cell epithelioma of the bladder in one patient were the underlying malignant diseases. Seven patients had a radical retropubic prostatectomy, two had radical retropubic prostatectomy after radiation, two had brachytherapy, and three were treated by a combination of radiation and brachytherapy. One patient formed a fistula after cystectomy and dilation of a stricture. This heterogenous group of patients received multiple therapies including initial colostomy (7 patients), transanal repair (2 patients), parasacral repair (2 patients), transperineal repair (2 patients), coloanal anastomosis (3 patients), and muscle transposition (3 patients). Four of our patients required a permanent stoma. CONCLUSION: In patients with iatrogenic rectourethral fistula that occurred after radical retropubic prostatectomy or radiation, fecal and urinary diversion and muscle transposition followed by re establishment of both urinary and intestinal continuity may be the treatment modality of choice. PMID- 10458121 TI - Glyceryl trinitrate ointment for the treatment of chronic anal fissure: results of a placebo-controlled trial and long-term follow-up. AB - PURPOSE: A randomized, double-blind, placebo-controlled trial was performed to test the effect of intra-anal glyceryl trinitrate ointment in patients with chronic anal fissures that would normally have been treated by sphincterotomy. Long-term follow-up was then performed to assess fissure healing. METHODS: Patients with chronic anal fissures were randomly assigned to 0.2 percent topical glyceryl trinitrate ointment or placebo. Anal manometry was performed before treatment, one week later, and 48 hours after treatment ceased at four weeks. Fissure healing was assessed by an observer blinded to the treatment arm. Pain was recorded on a linear analog scale. At the completion of the trial, treatment was continued with glyceryl trinitrate until fissure healing was obtained or lateral sphincterotomy was performed if required for ongoing pain. A long-term follow-up assessment was made at a mean of 29 (range, 25-33) months. RESULTS: There was a significant reduction in anal resting pressure at Week 1 with glyceryl trinitrate (P = 0.001) but not placebo, and at Week 4 there was a significant reduction in pain score with glyceryl trinitrate (P = 0.001) and placebo (P = 0.01) and a significant reduction in fissure grade with glyceryl trinitrate (P = 0.0001) and placebo (P = 0.02). Forty-six percent of fissures healed with glyceryl trinitrate and 16 percent healed with placebo (P = 0.001). At long-term follow-up in 40 of 43 patients, 14 patients (35 percent) had undergone lateral sphincterotomy, and in the remainder who were treated with glyceryl trinitrate there was a significant reduction in pain score (P = 0.0002). Seventeen patients attended for repeat manometry and fissures were healed with glyceryl trinitrate in ten (59 percent) cases. High internal sphincter pressures persisted at long-term follow-up in patients successfully treated with glyceryl trinitrate, indicating that the sphincter is the cause rather than effect of anal fissure. CONCLUSION: Topical glyceryl trinitrate produces a successful internal sphincterotomy, which resulted in long-term healing of 59 percent of chronic anal fissures and significant improvement in pain. Internal sphincter spasm is the cause of chronic anal fissure. PMID- 10458122 TI - Treatment of chronic anal fissure with topical glyceryl trinitrate. AB - PURPOSE: There has been an emerging trend to treat chronic anal fissures by pharmacologic means. In particular, by the topical application of glyceryl trinitrate. However, such treatment has failed to gain wide acceptance within our local surgical community. The aim of this study was to evaluate the efficacy and patient acceptance of topical glyceryl trinitrate in the management of chronic anal fissures. METHODS: A retrospective review was conducted on a consecutive series of 31 patients who were diagnosed as having a chronic anal fissures and were managed by the topical application of 0.2 percent glyceryl trinitrate to the perianal region twice a day. All patients were commenced on a high-fiber diet, and they were reviewed every two weeks until healed or until surgery was required. Patients were subsequently contacted by a mailed questionnaire to determine compliance with therapy, resolution of symptoms, recurrence of symptoms after cessation of therapy, and side effects. RESULTS: Of 31 patients, 27 completed the questionnaire. Only 67 percent (18/27) of patients complied with the treatment as prescribed, and just 56 percent (15/27) achieved complete resolution of their symptoms after three to ten weeks of therapy. Recurrence of symptoms developed in 27 percent (4/15) of these patients after a median follow up of six months. Side effects were reported in 78 percent (21/27) of patients, and in ten percent (2/21) they were so severe that the patients ceased therapy. CONCLUSION: In this study the treatment of chronic anal fissures with topical glyceryl trinitrate appeared less efficacious than previously reported. In addition, the study highlighted problems with patient compliance, healing rates, and side effects that are associated with this modality therapy. PMID- 10458123 TI - Nifedipine for local use in conservative treatment of anal fissures: preliminary results of a multicenter study. AB - PURPOSE: This study was performed according to a prospective, randomized, double blind, multicenter design. The aim was to test the efficacy of local application of nifedipine gel" in healing acute anal fissure by relaxing the internal anal sphincter. METHODS: Two hundred eighty-three patients who gave informed consent were recruited; they received a clinical examination. A questionnaire to evaluate the symptoms and the pain was administered, and a proctoscopy and anorectal manometry were performed. Patients treated with nifedipine (n = 141) used topical 0.2 percent nifedipine gel every 12 hours for three weeks. The control group, consisting of 142 patients, received topical 1 percent lidocaine and 1 percent hydrocortisone acetate gel during therapy. Manometry was performed before and on Days 14 and 21. Anal pressures were measured by recording resting and squeeze pressures. RESULTS: Results obtained were as follows: total remission from acute anal fissure was achieved after 21 days of therapy in 95 percent of the nifedipine-treated patients (P < 0.01), as opposed to 50 percent of the controls (P < 0.01), and previously elevated maximum resting anal pressures decreased from a mean value +/- standard deviation of 72.5 +/- 10.07 mmHg to 50.5 +/- 10.03 mmHg in the nifedipine group. This represents a mean reduction of 30 percent (P < 0.01). We also observed a significant decrease in squeeze pressures in nifedipine treated patients (from a mean +/- standard deviation of 130.5 +/- 19.25 mmHg to 108.5 +/- 18.55 mmHg, a mean reduction of 16.8 percent; P < 0.01). No changes in anal pressures were observed in the control group. We did not observe any systemic side effect or significant anorectal bleeding in patients treated with nifedipine. CONCLUSIONS: Our study clearly demonstrates that the therapeutic use of nifedipine, which at present is used only in cardiovascular pathologies, should be extended with local use to the conservative treatment of anal fissures. PMID- 10458124 TI - Therapeutic effects of different doses of botulinum toxin in chronic anal fissure. AB - PURPOSE: The aim of this study was to evaluate the clinical and manometric results of three different doses of botulinum toxin and two methods of injection for the treatment of chronic idiopathic anal fissure. METHODS: Sixty-nine patients with chronic anal fissure were included in a non-randomized, prospective trial of intrasphincteric injection of botulinum toxin. All patients reported postdefecatory anal pain lasting more than two months. Scoring systems were developed for anal pain, bleeding, and defecatory difficulty. Maximum resting and squeeze anal pressures were determined before and one month after treatment. Twenty-three patients undergoing a 5-U injection of diluted botulinum toxin A (BOTOX) on each side of the anal sphincter (total dose, 10 U) constituted the first group. In a second group 27 patients were injected as previously described, with an additional 5-U injection below the fissure (total dose, 15 U). The 19 patients constituting the third group received a 7-U injection on each side of the anus and below the fissure (total dose, 21 U). All patients were followed up for at least six months. RESULTS: Pain relief one month after treatment was more evident in the second and the third group (48 percent of patients in the first group, 74 percent in the second group, and 100 percent in the third group). A significant reduction of the mean resting pressure was demonstrated only in Groups II and III (P < 0.05), whereas the mean squeeze pressure significantly decreased in the three groups (P < 0.01 in Group I and P < 0.001 in Groups II and III). Fifty-two percent of the patients in the first group, 30 percent in the second group, and 37 percent in the third group were reinjected during the follow up period, because of persistence of symptomatology or early relapse. The need for surgery was similar in the first and the second group (17 and 19 percent, respectively) and clearly lower in the last group (5 percent). No serious complications or incontinence attributable to this therapeutic modality developed in any patient. CONCLUSIONS: Intrasphincteric injection of botulinum toxin is a reliable new option in the treatment of uncomplicated chronic anal fissure. The healing rate is related to the dose and probably to the number of puncture sites. No permanent damage to the continence mechanism was detected in these patients. PMID- 10458125 TI - Posterior sagittal anorectoplasty in adults: secondary repair for persistent incontinence in patients with anorectal malformations. AB - PURPOSE: Posterior sagittal anorectoplasty, regarded as a standard surgical primary repair for anorectal malformations in infancy, was evaluated for effectiveness when performed as a secondary operation for establishing continence in the adult. The purpose of this review was to evaluate our results of performing posterior sagittal anorectoplasty in adult patients and to emphasize the extensive evaluation required to perform proper patient selection. METHOD: From January 1, 1992, to December 31, 1996, eight patients with Grade 3 incontinence underwent posterior sagittal anorectoplasty. The ages ranged from 13 to 40 (mean, 26) years. RESULTS: All patients had diverting stomas at the time of repair and all but one had restoration of intestinal continuity. Of eight patients who underwent posterior sagittal anorectoplasty, one failed secondary to rectal ischemia and retained a diverting stoma. Six patients had restoration of continuity. Five patients were continent and one had incontinence only to gas. DISCUSSION: We have established that posterior sagittal anorectoplasty can effectively be used to establish continence as a secondary procedure for a select group of adult patients. PMID- 10458126 TI - Comparison of ileal pouch-anal anastomosis and ileorectal anastomosis in patients with familial adenomatous polyposis. AB - PURPOSE: The aim of this study was to evaluate the surgical complications and long-term outcome and assess the functional results and quality of life after ileorectal anastomosis and ileal pouch-anal anastomosis in patients with familial adenomatous polyposis. METHODS: From 1980 to 1997, 131 patients with familial adenomatous polyposis were operated on or were followed up or both at the Familial Gastrointestinal Cancer Registry at Mount Sinai Hospital. Demographic and operative data were prospectively collected in the ileal pouch-anal anastomosis group, and retrospectively in the ileorectal anastomosis group. A questionnaire or telephone interview or both were undertaken to evaluate functional outcome and quality of life. RESULTS: The ileorectal anastomosis group consisted of 60 patients (mean age, 31 years; mean follow-up, 7.7 years). In the ileal pouch-anal anastomosis group there were 50 patients (mean age, 35 years; mean follow-up, 6 years). There were no statistically significant differences with respect to anastomotic leak rate in ileal pouch-anal anastomosis vs. ileorectal anastomosis (12 vs. 3 percent; P = 0.21), risk of small-bowel obstruction (24 vs. 15 percent; P = 0.58), and risk of intra-abdominal sepsis (3 vs. 2 percent; P = 0.86). Reoperation rate was similar in the two groups (14 vs. 16 percent; P = 0.94). Twenty-one patients (37 percent) with ileorectal anastomosis were converted to ileal pouch-anal anastomosis (12 patients) or proctocolectomy (9 patients), because of rectal cancer (5 patients), dysplasia (1 patient), or uncontrollable rectal polyps (15 patients). Two pelvic pouches were excised, and another one was defunctioned. Information regarding functional results and quality of life was obtained in 40 patients (66.6 percent) in the ileorectal anastomosis group and in 43 patients (86 percent) in the ileal pouch anal anastomosis group. Patients with ileorectal anastomosis had a significantly better functional outcome with regard to nighttime continence and perineal skin irritation. But otherwise, functional results and quality of life were similar. CONCLUSIONS: Although ileorectal anastomosis has a better functional outcome, ileal pouch-anal anastomosis may be preferable because of the lower long-term failure rate. Ileorectal anastomosis is still an option in patients with familial adenomatous polyposis with rectal polyp sparing and good compliance for follow up. PMID- 10458127 TI - Abdominosacral resection of recurrent rectal cancer in the sacrum. AB - PURPOSE: Resection of the sacrum is the only curative therapy of isolated sacral recurrence after primarily resected rectal cancer. The aim of the study was to assess whether there is a benefit in terms of overall survival, morbidity, and mortality when sacrum resection is performed more radically and in cooperation between colorectal and orthopedic surgeons. Possible prognostic factors were also assessed. METHODS: Twelve consecutive patients who underwent interdisciplinary partial sacral resection were included in a retrospective cohort study. Furthermore, overall survival rate and survival time were calculated. RESULTS: Histologic examination showed tumor-free resection margins in all cases. Extended resection was necessary in seven patients, including total pelvic exenteration in two. No perioperative death occurred and no patient required early reoperation. Complications were observed in 42 percent of patients, mainly caused by poor wound healing. All patients experienced relief from pain. One-year and three-year overall survival rates were 50 and 17 percent, respectively. The overall mean survival time was 21.7 months. Patients who died of recurrent disease within one year either underwent former resection for locoregional recurrence, had extensive local recurrent tumors affecting pelvic visceral structures, or retrospectively suffered from metastatic sacral tumor manifestation. CONCLUSION: The mortality and morbidity rates observed in the present study seem to justify partial sacral resection as a means to achieve palliation of perineosacral pain in spite of rare overall long-term survival. PMID- 10458128 TI - Colorectal cancer in hereditary breast cancer kindreds. AB - PURPOSE: This study compared characteristics of colorectal cancer between families with dominant breast cancer inheritance and the general population. The cumulative incidence of colorectal cancer was also studied in genetically determined breast cancer syndrome subjects with BRCA1 and BRCA2 mutations and compared with the general population. METHODS: Subjects included 42 patients with colorectal cancer from 32 clinically determined hereditary breast cancer kindreds based on the autosomal dominant inheritance of breast cancers and early age of onset. The general population colorectal cancer cohort was composed of 755 patients from a tumor registry. Lifetime risk of colorectal cancer was determined in 164 BRCA1 and 88 BRCA2 gene mutation carriers and compared with the general population. Mean age of colorectal cancer onset, anatomic site distribution, histologic stage at presentation, and five year stage-stratified survival rates were compared between clinically determined hereditary breast cancer family members and the general population. RESULTS: The lifetime risk of colorectal cancer in male BRCA1 and BRCA2 mutation carriers was 5.6 percent, which was not different from 6 percent in males from the general population. Likewise, the lifetime colorectal cancer risk in female BRCA1 and BRCA2 mutation carriers was 3.2 percent, which was not different from 5.9 percent in females from the general population. Mean age of onset +/- standard error for patients with colorectal cancer was 60 +/- 2 years for hereditary breast cancer kindreds compared with 67 +/- 0.4 years for the general population (P = 0.0004). Colorectal cancer site distribution did not vary between hereditary breast cancer and the general population. Overall colorectal cancer stage distribution was significantly different, with more Stage I and fewer Stage IV cancers in subjects with hereditary breast cancer compared with the general population (P = 0.01). Overall five year stage-stratified colorectal cancer survival rate +/- standard error was 66 +/- 8 percent for hereditary breast cancer kindreds and 46 +/- 2 percent for the general population (P = 0.023). CONCLUSION: Lifetime cumulative colorectal cancer incidence in subjects with BRCA1 and BRCA2 gene mutations was not different from the general population. However, significant differences in colorectal cancer were noted between hereditary breast cancer family members and the general population. Hereditary breast cancer-associated colorectal cancer had an earlier age of onset, lower tumor stage, and better survival rate than the general population. Except for age of onset, colorectal cancer in hereditary breast cancer kindreds exhibited more favorable characteristics than colorectal cancer in the general population. PMID- 10458129 TI - Limitations with light microscopy in the detection of colorectal cancer cells. AB - PURPOSE: The failure of light microscopy to predict individual patient survival accurately in pStage I and II colorectal carcinoma can hinder planning postoperative therapy and follow-up. This study was designed and conducted in two parts to assess the influence of relative sensitivity of the light microscope on the pathologist's ability to detect malignant cells in lymph nodes. METHODS: The first part of the study examined the issue of sampling error as a fraction of the number of lymph node sections examined by asking the question, "Does increasing the number of sections (sampling) taken from the block increase tumor cell detection in a lymph node?" Three levels of five sections 4 to 5 microm thick separated by 15 to 20 microm were obtained from each of 494 blocks from 173 cases of pStage I and II colorectal carcinoma. A total of 1,721 lymph nodes were examined. Sections from each level were stained with hematoxylin and eosin and for the expression of cytokeratin. The second part of the study examined the relative sensitivity of the light microscope to detect tumor cells in a lymph node. To simulate lymph nodes, cell blocks were made that contained 10(6) or 10(7) mononuclear cells admixed with increasing numbers of SW480 tumor cells (0, 50, 10(2), 5 x 10(2), 10(3), and 5 x 10(3)). Three pathologists independently examined sections from ten control and ten experimental blocks. RESULTS: Results from the first part of the study demonstrated cytokeratin-positive cells in 278 lymph nodes from 102 of 172 (59 percent) cases. These cells were identified in the first level in 177 (64 percent) as compared with the second or third level or both in 101 (36 percent) of the lymph nodes. Results from the second part of the study demonstrated an overall sensitivity of light microscopic examination of hematoxylin and eosin-stained sections to be approximately 23 percent, representing tumor cells correctly detected in 7 sections of the 30 sections containing tumor cells. The overall specificity was 87 percent or 26 sections correctly classified as lacking tumor cells of a possible 30. Immunohistochemical staining for cytokeratin expression improved sensitivity of the light microscope to detect tumor cells to 18 of 30 (60 percent) and the specificity to 30 of 30 (100 percent). CONCLUSION: This study demonstrates several sources of variability that can induce errors in pathologic staging. These include 1) inadequate section, i.e., sampling, of lymph nodes, 2) use of only hematoxylin and eosin stained sections, 3) samples with tumor cells below the level of detection sensitivity of the light microscope, and 4) observer variability. PMID- 10458130 TI - Histopathologic characteristics of colorectal cancer with liver metastasis. AB - PURPOSE: Although prognostic factors of colorectal cancer have been studied, factors associated with liver metastasis have not been fully investigated. The aim of this study was to clarify the histopathologic characteristics of colorectal cancer with liver metastasis. METHODS: We performed a retrospective histopathologic study on 335 patients who underwent resection of colorectal cancer during 15 years. Histopathologic parameters of tumors with liver metastasis were compared with those without liver metastasis. RESULTS: Forty-one patients (12 percent) had simultaneous liver metastasis. Tumors having liver metastasis, when compared with those not having liver metastasis, were characterized by high frequency of tumor size more than 6 cm (51 vs. 28 percent; P < 0.01), presence of serosal invasion (98 vs. 66 percent; P < 0.01), lymphatic invasion (34 vs. 15 percent; P < 0.01), venous invasion (24 vs. 3 percent; P < 0.01), and lymph node metastasis (85 vs. 39 percent; P < 0.01). Multivariate analysis showed that factors independently associated with liver metastasis were serosal invasion, venous invasion, and lymph node metastasis. Accuracy in the diagnosis of liver metastasis was highest for venous invasion (88 percent) and lowest for serosal invasion (41 percent). Among 98 patients with both serosal invasion and lymph node metastasis, tumors with and without liver metastasis were different in frequency of venous invasion (26 vs. 6 percent; P < 0.01) and extracolic lymph node metastasis (68 vs. 47 percent; P < 0.05). CONCLUSION: In colorectal cancer important factors associated with liver metastasis were serosal invasion, venous invasion, and lymph node metastasis. Significant determinants for liver metastasis from colorectal cancer were venous invasion and extracolic lymph node metastasis. PMID- 10458131 TI - Use of endoanal ultrasound in patients with rectovaginal fistulas. AB - PURPOSE: The purpose of our study was to define the role of endoanal ultrasound in the evaluation and management of patients with rectovaginal fistula. METHODS: A retrospective review was performed of all patients with rectovaginal fistula who were evaluated by endoanal ultrasound at Barnes-Jewish Hospital at Washington University from 1992 to 1997. RESULTS: Twenty-five females underwent endoanal ultrasound before rectovaginal fistula repair. Mean age was 34 years. Rectovaginal fistulas were caused by obstetric trauma (19 patients; 76 percent), cryptoglandular disease (5 patients; 20 percent), and Crohn's disease (1 patient; 4 percent). Previous rectovaginal fistula repair had been performed in ten patients (40 percent). A history of anal incontinence was present in ten patients (40 percent). Rectovaginal fistula location was above (15 patients), at (7 patients), or below (3 patients) the dentate line. Rectovaginal fistula size was <5 mm (19 patients; 76 percent) or >5 mm (6 patients; 24 percent). Anal manometry revealed decreased sphincter pressures (resting or squeeze) in 12 patients (48 percent). Pudendal nerve latency was abnormal in three patients (9 percent). Endoanal ultrasound identified the rectovaginal fistula in 7 patients (28 percent) and an anterior sphincter defect in 23 patients (92 percent). At surgery sphincter injuries were identified in 23 patients (92 percent). Treatment was either sliding flap repair with anal sphincter reconstruction (22 patients; 88 percent) or sliding flap repair alone (3 patients; 12 percent). Repair of the rectovaginal fistula was successful in 23 patients (92 percent). Complications occurred in 11 patients (44 percent): two recurrent rectovaginal fistulas, five infections, two skin separations, one ectropion, and one hematoma. The two patients with recurrent rectovaginal fistula had prior repairs, and both were subsequently repaired successfully. Of the 11 patients with preoperative anal incontinence, 6 patients (54 percent) were continent and 2 (18 percent) improved after surgery. Cause, size, location, and previous repair of fistula had no effect on final outcome. CONCLUSIONS: Noncontrast endoanal ultrasound was not useful in imaging rectovaginal fistulas and cannot be recommended as a diagnostic or screening tool for the identification of a rectovaginal fistula. However, we recommend that endoanal ultrasound be performed preoperatively in all patients with known rectovaginal fistulas to identify and map occult sphincter defects. Concomitant anal sphincter reconstruction should be considered strongly in patients with rectovaginal fistula and an endoanal ultrasound-documented sphincter defect. PMID- 10458132 TI - Influence of erythropoietin on transfusion requirements in patients receiving preoperative chemoradiotherapy for rectal cancer. AB - PURPOSE: Perioperative homologous blood transfusion has been suggested to have an adverse effect on survival in patients undergoing resection of colorectal cancers. Preoperative therapy is being increasingly used for rectal cancer patients and has an adverse effect on erythropoietic capacity. The objectives of this study were to evaluate the feasibility and safety of administration of recombinant human erythropoietin to patients receiving preoperative therapy for rectal cancer and to assess the impact of such treatment on blood transfusion requirements. METHODS: The study was an open-label, Phase I and II, nonrandomized, two-center trial. All patients received 50.4 Gy of irradiation with 5-fluorouracil infusions. Ten patients diagnosed with rectal cancer received 250 U/kg of recombinant human erythropoietin subcutaneously three times per week during preoperative radiation and chemotherapy. Oral iron was given to patients receiving erythropoietin. Ten contemporaneously treated patients who received both radiation and chemotherapy were used as controls. RESULTS: Of the 20 patients 13 were males; mean age was 64 years. Surgical procedures that patients underwent were abdominoperineal resection (14 patients), low anterior resection (4 patients), coloanal anastomosis (1 patient), or none (1 patient). There were no significant differences between groups in age, gender, stage or hemoglobin levels before therapy. No adverse reactions to erythropoietin were encountered. Hemoglobin levels were significantly higher in the treatment group during Weeks 1, 3, and 5 (P < 0.02 for each). Transfusion requirements were significantly decreased in patients who received erythropoietin (0.4 vs. 3.7 units; P < 0.0003). CONCLUSIONS: The data showed that use of erythropoietin during preoperative therapy can prevent the decline in hemoglobin that commonly occurs during therapy. Further, this was not associated with adverse events and significantly decreased the need for perioperative blood transfusions. This suggests that the use of erythropoietin in support of a preoperative chemoradiotherapy regimen for patients with rectal cancer is safe and should be considered. Whether such transfusion avoidance will translate into a survival benefit in this setting will require a large, prospective, clinical trial. PMID- 10458133 TI - Monitoring and antibacterial treatment for postoperative bacterial overgrowth in Crohn's disease. AB - PURPOSE: Bacterial overgrowth sometimes complicates the clinical course of Crohn's disease and may lead to inappropriate treatment. To clarify the effect of antibiotic therapy, we monitored the hydrogen concentration in expiratory breath after fasting. METHODS: We evaluated 18 patients (15 males; median age, 32.7; range, 22.3-60 years) for postoperative bacterial overgrowth symptoms and for intestinal dilation by plain abdominal x-ray. Five patients had ileitis and 13 patients had ileocolitis. Various intestinal resections were performed in all, and strictureplasties were done at the same time in 13 patients. The median postoperative period was 10.2 (range, 1.2-102) months. Nine patients, who had symptoms such as bloating, nausea, vomiting, or pain, were classified as the symptomatic group, whereas nine other patients, who had no symptoms, were classified as the symptom-free group. Sixteen patients who had undergone intestinal resections for noninflammatory bowel disease served as the control group. After overnight fasting, hydrogen concentration in end-expiration, breath was measured with gas chromatography. At the same time clinical examinations of white blood cell count, hemoglobin, total protein, serum albumin, iron, sialic acid, and C-reactive protein in the peripheral blood were performed. To assess the effect of antibacterial treatment, changes in symptoms were assessed in eight patients who received antibacterial treatment. Hydrogen concentration was measured repeatedly before and after treatment in six patients. RESULTS: The symptomatic group had an expiratory hydrogen concentration level significantly higher (median, 40; range, 20-139 ppm) than the control group (median, 3; range, 1-6 ppm) and the symptom-free group (median, 4; range, 1-10 ppm). After the antibiotic treatment the symptoms were improved in all of the patients, and the hydrogen concentration level was significantly reduced (median, 4.5; range, 2-13 ppm). CONCLUSIONS: Antibacterial treatment was useful in the postoperative patients whose assessments were complicated by bacterial overgrowth. Using a hydrogen breath test, bacterial overgrowth was effectively monitored and managed, effecting a change in clinical symptoms. PMID- 10458134 TI - Attenuated adenomatous polyposis coli: the role of ascertainment bias through failure to dye-spray at colonoscopy. AB - PURPOSE: The aim of this study is to show that the diagnosis of attenuated adenomatous polyposis coli must be made with caution and certainly only after adequate colonic examination with dye-spray. METHODS: Four patients thought to have attenuated adenomatous polyposis coli on the basis of family history and the identification of fewer than 100 polyps on simple colonoscopy underwent colonoscopy with dye-spray. RESULTS: All four individuals were found to have more than 100 polyps when dye-spray was used, confirming a diagnosis of classical familial adenomatous polyposis. CONCLUSIONS: The diagnosis of familial adenomatous polyposis may be missed altogether or incorrectly assigned as attenuated adenomatous polyposis coli if dye-spray is not used at colonoscopy. Patients with a family history of familial adenomatous polyposis or colorectal cancer should be considered for dye-spray before the diagnosis of familial adenomatous polyposis is excluded or one of attenuated adenomatous polyposis coli is made. PMID- 10458135 TI - Sigmoid volvulus: study from a north Indian hospital. AB - PURPOSE: The aim of this study was to review our experience with sigmoid volvulus and to compare the various surgical procedures performed for treatment of sigmoid volvulus. METHODS: A retrospective study of 111 patients with sigmoid volvulus treated at University Hospital in northern India during last six years was performed. RESULTS: Mean age for sigmoid volvulus was 51.1 +/- 15.3 years, with a male-to-female ratio of 2.5:1. Duration of symptoms was 4.3 +/- 2.6 days. Plain radiographs of the abdomen suggested the diagnosis in 76 cases (68.47 percent). Resection of the bowel was performed in 40 cases with gangrenous sigmoid colon and 6 cases with viable colon. Nonresectional procedure in the form of colocolopexy (fixation of sigmoid to transverse colon) or extraperitonealization was performed in 13 and 44 cases, respectively. Mortality was 6.3 percent (n = 7). No recurrence was seen after resection and extraperitonealization, whereas 5 of 13 cases (38.5 percent) had recurrence after colocolopexy. CONCLUSION: Resection and extraperitonealization of the sigmoid colon are the suitable alternatives for nongangrenous sigmoid volvulus. Colocolopexy is not indicated, because of the associated high recurrence rate. PMID- 10458136 TI - Benefits of endorectal ultrasound for management of smooth-muscle tumor of the rectum: report of three cases. AB - Smooth-muscle tumor of the rectum is rare, and the therapeutic strategy is still controversial. Endorectal ultrasound was used to evaluate three patients with smooth-muscle tumor of the rectum. Endorectal ultrasound demonstrated a homogenous hypoechoic tumor without invasion to the perirectal tissue in two patients. The tumor was 5 cm in diameter in one patient and 4 cm in diameter in the other patient, and they were excised locally. Their histologic types were leiomyoma and leiomyosarcoma. The third patient had a recurrent leiomyosarcoma. Proctosigmoidoscopy found a linear lesion with ulcerated mucosa on the rectal wall. Endorectal ultrasound observed a hypoechoic solid tumor of 3.5 cm x 1 cm, which involved the mucosal, submucosal, and muscle layers of the rectal wall. Disruption of the first hypoechoic layer was identified. Abdominoperineal resection was performed. Endorectal ultrasound follow-up revealed no evidence of recurrence in any of these patients. Endorectal ultrasound can help to define the extent of disease and may be a useful adjunct in deciding about the appropriate surgical procedure in these diseases. PMID- 10458137 TI - Adenosquamous carcinoma of the rectum showing endocrine-cell differentiation: report of a case. AB - PURPOSE: Previous records of adenosquamous carcinomas with endocrinologic features rarely have been reported. Although the disease behaves in an extremely aggressive manner, chemotherapy after surgery has never been proposed. We used regional chemotherapy for treatment of unresectable liver metastases. METHODS: Hartmann colostomy was performed and 5-fluorouracil was infused into the hepatic artery for 15 weeks after the operation. RESULTS: Multiple liver metastases were present at the initial operation. Three months after the start of the chemotherapy, computed tomography showed that metastatic tumors in the liver had disappeared. The patient survived 18 months after the initial operation. CONCLUSION: This case report describes the first successful treatment with adjuvant regional chemotherapy of a patient who had an adenosquamous carcinoma with endocrine-cell differentiation in the rectum and liver metastases. PMID- 10458138 TI - Refractory proctosigmoiditis caused by myointimal hyperplasia of mesenteric veins: report of a case. AB - PURPOSE: Proctosigmoiditis occurring in young adults is usually idiopathic and usually responds to medical management. If the process progresses to pancolitis and is refractory to medical management, proctocolectomy may be required. Myointimal hyperplasia of mesenteric veins, though rare, may also cause proctosigmoiditis, but this entity, in contrast to the idiopathic variety, does not respond to medical management; surgical excision limited to the involved colonic segment is curative. Because the treatment of the two entities differs significantly, it is important to distinguish them diagnostically. The purpose of this case report is to increase awareness of myointimal hyperplasia of mesenteric veins and to emphasize the clinical features that distinguish it from idiopathic proctosigmoiditis. METHODS: We report the case of a twenty-two-year-old male with an inflammatory process involving the distal colon and rectum, initially thought to be idiopathic proctosigmoiditis. The inflammation did not respond to an extensive course of medical management, and the patient developed complications associated with both the disease process and his medical therapy. Surgical resection of the rectosigmoid was performed. RESULTS: Histologic examination of the resected colon revealed the underlying process to be colonic ischemia caused by myointimal hyperplasia of mesenteric veins not associated with idiopathic inflammatory bowel disease or systemic vasculitis. CONCLUSION: Proctosigmoiditis caused by myointimal hyperplasia of mesenteric veins and idiopathic proctosigmoiditis may present in a similar fashion. Although patients with myointimal hyperplasia of mesenteric veins do not respond to medical management, segmental resection is usually curative, and long-term drug therapy or even proctocolectomy can be avoided. Physicians should consider the possibility of myointimal hyperplasia of mesenteric veins when patients with apparent idiopathic proctosigmoiditis do not respond to medical therapy. PMID- 10458139 TI - Total anorectal and partial vaginal reconstruction with dynamic graciloplasty and colonic vaginoplasty after extended abdominoperineal resection: report of a case. AB - PURPOSE: Quality of life is altered after abdominoperineal resection, because of permanent iliac colostomy. Psychological rehabilitation is even more difficult after extended abdominoperineal resection to the vagina, because of the loss of both continence and sexual functions. We report the first case of total anorectal and vaginal reconstruction using dynamic graciloplasty and colonic vaginoplasty after extended abdominoperineal resection. METHODS: A 46-year-old female underwent extended abdominoperineal resection with posterior colpectomy for a low rectal adenocarcinoma infiltrating the anal sphincter and vagina. Anorectal reconstruction was performed with coloperineal anastomosis and double dynamic graciloplasty. Vaginal reconstruction was performed using a 10-cm, isolated, rotated sigmoid loop. The procedure was performed in three stages, including abdominoperineal resection with reconstruction, implantation of the stimulator, and closure of the temporary ileostomy. RESULTS: Resting and electrostimulated pressures of the neosphincter were 40 and 110 cm H2O respectively. Continence was achieved for formed stools two months after closure of the stoma, with spontaneous defecations (30-90 minutes). The patient experienced regular sexual activity six months after closure of the stoma. CONCLUSION: This new original technique can be proposed in selected young females after extended abdominoperineal resection, to preserve continence, sexual activity, and body image. PMID- 10458140 TI - Treatment of external anorectal mucosal prolapse with circular stapler: an easy and effective new surgical technique. AB - PURPOSE: The aim of this study was to demonstrate the feasibility, effectiveness, and reliability of a new technique for treating overt rectal mucosal prolapse using a stapler device. METHODS: Eighteen consecutive patients with overt rectal mucosal prolapse were selected for the study. Preoperative anal manometry and cinedefecography demonstrated no anal incontinence and the absence of full thickness rectal prolapse. One or two purse strings were prepared 3 to 4 cm distally to the dentate line and tied on a 33 mm circular stapler introduced through the anus and then fired. RESULTS: The operation lasted an average of 15 minutes, and no local complications were recorded. Supplementary hemostatic sutures (2 to 6 stitches) were sometimes necessary. Seven patients did not require postoperative analgesia, whereas eight patients received one or two administrations of analgesics. Longer-lasting analgesic treatment (4 days) was necessary in only three patients. Sixteen patients were discharged after 48 hours, and only 1 after four days because of pulmonary infection. Patients resumed normal activities after a median period of three days. Median follow-up was 20 months. The prolapse was eliminated in all cases. No stricture was found at anal exploration, and no episodes of anal incontinence or bleeding were recorded. Postoperative manometry did not show significant changes compared with preoperative findings. CONCLUSIONS: This new surgical technique is safe, effective, and rapid, causing minimum or no postoperative pain and could be proposed to replace traditional surgery for this common condition. PMID- 10458141 TI - Headaches in the treatment of anal fissure. PMID- 10458142 TI - Academic titles and authorship. PMID- 10458143 TI - Lymph node recovery from colorectal resection specimens. PMID- 10458144 TI - Researcher sues university over rights to transgenic technology. PMID- 10458145 TI - Coordinated work reveals plant evolution. PMID- 10458146 TI - 'Strengthened' Icelandic bioethics committee comes under fire. PMID- 10458147 TI - UK space strategy sets sights on Mars. PMID- 10458148 TI - Legal threat in bid to ban xenotransplants. PMID- 10458149 TI - US acts to lengthen term of patents to help biotechnology. PMID- 10458150 TI - Japan plans to label 'detectable' GM food. PMID- 10458151 TI - Animal rights activists attack Gore over chemical screening. PMID- 10458152 TI - Endocrine disrupter verdict left open. PMID- 10458153 TI - Why quasi-persons are not patentable. PMID- 10458154 TI - Where a little aid could go a long way. PMID- 10458155 TI - Developing B-cell theories. PMID- 10458156 TI - Evolution. Creatures from another world. PMID- 10458157 TI - Oscillations in the basal ganglia. PMID- 10458158 TI - Convergent evolution of cytokines. PMID- 10458159 TI - Photon-stimulated desorption as a substantial source of sodium in the lunar atmosphere. AB - Mercury and the Moon both have tenuous atmospheres that contain atomic sodium and potassium. These chemicals must be continuously resupplied, as neither body can retain the atoms for more than a few hours. The mechanisms proposed to explain the resupply include sputtering of the surface by the solar wind, micrometeorite impacts, thermal desorption and photon-stimulated desorption. But there are few data and no general agreement about which processes dominate. Here we report laboratory studies of photon-stimulated desorption of sodium from surfaces that simulate lunar silicates. We find that bombardment of such surfaces at temperatures of approximately 250 K by ultraviolet photons (wavelength lambda < 300 nm) causes very efficient desorption of sodium atoms, induced by electronic excitations rather than by thermal processes or momentum transfer. The flux at the lunar surface of ultraviolet photons from the Sun is sufficient to ensure that photon-stimulated desorption of sodium contributes substantially to the Moon's atmosphere. On Mercury, solar heating of the surface implies that thermal desorption will also be an important source of atmospheric sodium. PMID- 10458160 TI - Genome complexity, robustness and genetic interactions in digital organisms. AB - Digital organisms are computer programs that self-replicate, mutate and adapt by natural selection. They offer an opportunity to test generalizations about living systems that may extend beyond the organic life that biologists usually study. Here we have generated two classes of digital organism: simple programs selected solely for rapid replication, and complex programs selected to perform mathematical operations that accelerate replication through a set of defined 'metabolic' rewards. To examine the differences in their genetic architecture, we introduced millions of single and multiple mutations into each organism and measured the effects on the organism's fitness. The complex organisms are more robust than the simple ones with respect to the average effects of single mutations. Interactions among mutations are common and usually yield higher fitness than predicted from the component mutations assuming multiplicative effects; such interactions are especially important in the complex organisms. Frequent interactions among mutations have also been seen in bacteria, fungi and fruitflies. Our findings support the view that interactions are a general feature of genetic systems. PMID- 10458161 TI - Dynamics of disease resistance polymorphism at the Rpm1 locus of Arabidopsis. AB - The co-evolutionary 'arms race' is a widely accepted model for the evolution of host-pathogen interactions. This model predicts that variation for disease resistance will be transient, and that host populations generally will be monomorphic at disease-resistance (R-gene) loci. However, plant populations show considerable polymorphism at R-gene loci involved in pathogen recognition. Here we have tested the arms-race model in Arabidopsis thaliana by analysing sequences flanking Rpm1, a gene conferring the ability to recognize Pseudomonas pathogens carrying AvrRpm1 or AvrB. We reject the arms-race hypothesis: resistance and susceptibility alleles at this locus have co-existed for millions of years. To account for the age of alleles and the relative levels of polymorphism within allelic classes, we use coalescence theory to model the long-term accumulation of nucleotide polymorphism in the context of the short-term ecological dynamics of disease resistance. This analysis supports a 'trench warfare' hypothesis, in which advances and retreats of resistance-allele frequency maintain variation for disease resistance as a dynamic polymorphism. PMID- 10458162 TI - Time-dependent reorganization of brain circuitry underlying long-term memory storage. AB - Retrograde amnesia observed following hippocampal lesions in humans and animals is typically temporally graded, with recent memory being impaired while remote memories remain intact, indicating that the hippocampal formation has a time limited role in memory storage. However, this claim remains controversial because studies involving hippocampal lesions tell us nothing about the contribution of the hippocampus to memory storage if this region was present at the time of memory retrieval. We therefore used non-invasive functional brain imaging using (14C)2-deoxyglucose uptake to examine how the brain circuitry underlying long term memory storage is reorganized over time in an intact brain. Regional metabolic activity in the brain was mapped in mice tested at different times for retention of a spatial discrimination task. Here we report that increasing the retention interval from 5 days to 25 days resulted in both decreased hippocampal metabolic activity during retention testing and a loss of correlation between hippocampal metabolic activity and memory performance. Concomitantly, a recruitment of certain cortical areas was observed. These results indicate that there is a time-dependent reorganization of the neuronal circuitry underlying long-term memory storage, in which a transitory interaction between the hippocampal formation and the neocortex would mediate the establishment of long lived cortical memory representations. PMID- 10458163 TI - Memory for places learned long ago is intact after hippocampal damage. AB - The hippocampus is part of a system of structures in the medial temporal lobe that are essential for memory. One influential view of hippocampal function emphasizes its role in the acquisition and retrieval of spatial knowledge. By this view, the hippocampus constructs and stores spatial maps and is therefore essential for learning and remembering places, including those learned about long ago. We tested a profoundly amnesic patient (E.P.), who has virtually complete bilateral damage to the hippocampus and extensive damage to adjacent structures in the medial temporal lobe. We asked him to recall the spatial layout of the region where he grew up, from which he moved away more than 50 years ago. E.P. performed as well as or better than age-matched control subjects who grew up in the same region and also moved away. In contrast, E.P. has no knowledge of his current neighbourhood, to which he moved after he became amnesic. Our results show that the medial temporal lobe is not the permanent repository of spatial maps, and support the view that the hippocampus and other structures in the medial temporal lobe are essential for the formation of long-term declarative memories, both spatial and non-spatial, but not for the retrieval of very remote memories, either spatial or non-spatial. PMID- 10458164 TI - A basal ganglia pacemaker formed by the subthalamic nucleus and external globus pallidus. AB - The subthalamic nucleus of the basal ganglia (STN) is important for normal movement as well as in movement disorders. Lesioning or deep-brain stimulation of the STN can alleviate resting tremor in Parkinson's disease. The STN and its target nuclei display synchronized oscillatory burst discharge at low frequencies, some of which correlate with tremor, but the mechanism underlying this synchronized bursting is unknown. Here we show that the excitatory STN and inhibitory, external globus pallidus (GPe) form a feedback system that engages in synchronized bursting. In mature organotypic cortex-striatum-STN-GPe cultures, neurons in the STN and GPe spontaneously produce synchronized oscillating bursts at 0.4, 0.8 and 1.8 Hz. Pallidal lesion abolishes this bursting, whereas cortical lesion favours bursting at 0.8 Hz. Pallidal bursts, although weaker than STN bursts, were required for synchronized oscillatory burst generation by recruitment of subthalmic rebound excitation. We propose that the STN and GPe constitute a central pacemaker modulated by striatal inhibition of GPe neurons. This pacemaker could be responsible for synchronized oscillatory activity in the normal and pathological basal ganglia. PMID- 10458165 TI - Continued RAG expression in late stages of B cell development and no apparent re induction after immunization. AB - Models of B-cell development in the immune system suggest that only those immature B cells in the bone marrow that undergo receptor editing express V(D)J recombination-activating genes (RAGs). Here we investigate the regulation of RAG expression in transgenic mice carrying a bacterial artificial chromosome that encodes a green fluorescent protein reporter instead of RAG2. We find that the reporter is expressed in all immature B cells in the bone marrow and spleen. Endogenous RAG messenger RNA is expressed in immature B cells in bone marrow and spleen and decreases by two orders of magnitude as they acquire higher levels of surface immunoglobulin M (IgM). Once RAG expression is stopped it is not re induced during immune responses. Our findings may help to reconcile a series of apparently contradictory observations, and suggest a new model for the mechanisms that regulate allelic exclusion, receptor editing and tolerance. PMID- 10458166 TI - A SMAD ubiquitin ligase targets the BMP pathway and affects embryonic pattern formation. AB - The TGF-beta superfamily of proteins regulates many different biological processes, including cell growth, differentiation and embryonic pattern formation. TGF-beta-like factors signal across cell membranes through complexes of transmembrane receptors known as type I and type II serine/threonine-kinase receptors, which in turn activate the SMAD signalling pathway. On the inside of the cell membrane, a receptor-regulated class of SMADs are phosphorylated by the type-I-receptor kinase. In this way, receptors for different factors are able to pass on specific signals along the pathway: for example, receptors for bone morphogenetic protein (BMP) target SMADs 1, 5 and 8, whereas receptors for activin and TGF-beta target SMADs 2 and 3. Phosphorylation of receptor-regulated SMADs induces their association with Smad4, the 'common-partner' SMAD, and stimulates accumulation of this complex in the nucleus, where it regulates transcriptional responses. Here we describe Smurf1, a new member of the Hect family of E3 ubiquitin ligases. Smurf1 selectively interacts with receptor regulated SMADs specific for the BMP pathway in order to trigger their ubiquitination and degradation, and hence their inactivation. In the amphibian Xenopus laevis, Smurf1 messenger RNA is localized to the animal pole of the egg; in Xenopus embryos, ectopic Smurf1 inhibits the transmission of BMP signals and thereby affects pattern formation. Smurf1 also enhances cellular responsiveness to the Smad2 (activin/TGF-beta) pathway. Thus, targeted ubiquitination of SMADs may serve to control both embryonic development and a wide variety of cellular responses to TGF-beta signals. PMID- 10458167 TI - Trigger factor and DnaK cooperate in folding of newly synthesized proteins. AB - The role of molecular chaperones in assisting the folding of newly synthesized proteins in the cytosol is poorly understood. In Escherichia coli, GroEL assists folding of only a minority of proteins and the Hsp70 homologue DnaK is not essential for protein folding or cell viability at intermediate growth temperatures. The major protein associated with nascent polypeptides is ribosome bound trigger factor, which displays chaperone and prolyl isomerase activities in vitro. Here we show that delta tig::kan mutants lacking trigger factor have no defects in growth or protein folding. However, combined delta tig::kan and delta dnaK mutations cause synthetic lethality. Depletion of DnaK in the delta tig::kan mutant results in massive aggregation of cytosolic proteins. In delta tig::kan cells, an increased amount of newly synthesized proteins associated transiently with DnaK. These findings show in vivo activity for a ribosome-associated chaperone, trigger factor, in general protein folding, and functional cooperation of this protein with a cytosolic Hsp70. Trigger factor and DnaK cooperate to promote proper folding of a variety of E. coli proteins, but neither is essential for folding and viability at intermediate growth temperatures. PMID- 10458168 TI - Macular translocation for subfoveal choroidal neovascularization in age-related macular degeneration: a prospective study. AB - PURPOSE: To conduct a prospective study of macular translocation in patients with subfoveal choroidal neovascularization secondary to age-related macular degeneration. METHODS: In 10 eyes of 10 patients with subfoveal choroidal neovascularization and best-corrected visual acuity ranging from 20/50 to 20/800 (median, 20/111), the fovea was relocated by means of scleral imbrication, intentional retinal detachment with small posterior retinotomies, and partial fluid-air exchange. In two eyes, the choroidal neovascular membranes were removed at the time of macular translocation; in seven eyes they were photocoagulated in the postoperative period; and in one eye the membrane was removed during reoperation to unfold a macular fold. RESULTS: All 10 eyes were followed up for 6 months. The median postoperative foveal displacement was 1286 microm (range, 114 to 1,919 microm). In three eyes (30%), a foveal fold formed postoperatively requiring reoperation, with one of these eyes requiring a second reoperation for a rhegmatogenous retinal detachment. Best-corrected visual acuity improved in four eyes (median, 10.5 letters) and decreased in six eyes (median, 14.5 letters). The median change in visual acuity was a decrease of 5 letters. The final best-corrected visual acuity was 20/80 in two eyes, 20/126 in one eye, 20/160 in four eyes, 20/200 in one eye, 20/250 in one eye, and 20/640 in one eye. CONCLUSIONS: Our initial experience with limited macular translocation suggests that this surgical technique is unpredictable. However, in patients with subfoveal choroidal neovascularization from age-related macular degeneration, it offers the potential for improving visual function and may be associated with less loss of vision than the disease itself, if allowed to progress. Further refinements in surgical indications and technique are needed to make this procedure safer, more predictable, and more beneficial. PMID- 10458169 TI - Surgical treatment of submacular hemorrhage associated with idiopathic polypoidal choroidal vasculopathy. AB - PURPOSE: To report the visual outcome of surgical treatment of submacular hemorrhage associated with idiopathic polypoidal choroidal vasculopathy. METHODS: Eight eyes of eight consecutive patients with thick submacular hemorrhages associated with idiopathic polypoidal choroidal vasculopathy were treated with pars plana vitrectomy and tissue plasminogen activator-assisted removal of subretinal blood (December 1995 to September 1997) or intravitreal 100% sulfur hexafluoride gas injection without tissue plasminogen activator (October 1997 to March 1998). RESULTS: Postoperatively, laser treatment was performed for active polypoidal lesions outside the foveal avascular zone in four eyes. A retinal pigment epithelial tear was seen outside the foveal avascular zone in three eyes, and one eye developed a retinal detachment. The best-corrected visual acuity improved (by 3 or more lines) or stabilized in seven of the eight eyes. Four eyes had a final best-corrected visual acuity of 20/40 or better, and three eyes had a final best-corrected visual acuity of 20/50 to 20/200. In one eye, the visual acuity decreased from 20/100 to 20/500 because of the development of a subfoveal neovascular membrane. The membrane was excised, and histologic examination showed fibrovascular tissue between the retina and retinal pigment epithelium (type 2 pattern). CONCLUSIONS: Surgical intervention may be of benefit in eyes with submacular hemorrhage associated with idiopathic polypoidal choroidal vasculopathy. PMID- 10458170 TI - Limiting photoreceptor death and deconstruction during experimental retinal detachment: the value of oxygen supplementation. AB - PURPOSE: To assess the role of hypoxia in causing the death and deconstruction of photoreceptors in detached retinas and the effectiveness of supplemental oxygen in limiting such damage. METHODS: Retinal detachment was induced surgically in the right eye of each of 10 cats. The cats were allowed to survive surgery for 3 days. Two were kept for these 3 days in normoxia (room air, 21% oxygen) and eight in hyperoxia (70% oxygen). The retinas were examined for cell death by use of labels for normal and fragmenting DNA, with antibodies and a cone sheath-specific lectin to demonstrate the status of their inner and outer segments, the synaptic structures of the outer plexiform layer, and the distribution of basic fibroblast growth factor (bFGF) and with in situ hybridization to demonstrate bFGF mRNA. RESULTS: Retinal detachment without oxygen supplementation caused the death of some photoreceptors; the loss of cytochrome oxidase from the inner segments and the collapse of the outer segments of surviving photoreceptors; the loss of synaptophysin profiles from the outer plexiform layer; and the loss of bFGF protein from retinal neurons and neuroglia but not from retinal vessels. Oxygen supplementation (hyperoxia) during detachment mitigated all these changes, reducing photoreceptor death, maintaining the specialized structures of surviving photoreceptors, and stabilizing the bFGF within the retina. CONCLUSIONS: In experimental retinal detachment, hypoxia caused by the separation of outer retina from its normal source of nutrients is a factor in inducing the death and deconstruction of photoreceptors as well as in the loss of bFGF from the detached retina. Hyperoxia offered to human patients between diagnosis of retinal detachment and surgery may enhance the function of the reattached retina. PMID- 10458171 TI - Limiting the proliferation and reactivity of retinal Muller cells during experimental retinal detachment: the value of oxygen supplementation. AB - PURPOSE: To assess the role of hypoxia in inducing the proliferation, hypertrophy, and dysfunction of Muller cells in detached retina and the effectiveness of supplemental oxygen in limiting these reactions. METHODS: Retinal detachments were produced in the right eye of each of 13 cats; the cats survived surgery for 3 days, during which six were kept in normoxia (room air, 21%) and seven in hyperoxia (70% oxygen). Retinas were labeled for proliferation with an antibody (MIB-1) to a cell cycle protein (Ki-67), for evidence of hypertrophy employing antibodies to the intermediate filament protein glial fibrillary acidic protein (GFAP) and to beta-tubulin and for disturbance of glutamate neurochemistry employing antibodies to glutamate to a glutamate receptor (GluR-2) and to glutamine synthetase. RESULTS: Results from the two animals kept in normoxia after retinal detachment confirmed previous reports that detachment caused the proliferation of Muller cells, the hypertrophy of Muller cell processes, and the disruption of glutamate recycling by Muller cells. Oxygen supplementation during detachment reduced Muller cell proliferation and hypertrophy and reduced the abnormalities in the distributions of glutamate, GluR 2, and glutamine synthetase. CONCLUSIONS: Oxygen supplementation reduced the reaction of retinal Muller cells to retinal detachment, limiting their proliferation and helping to maintain their normal structure and function. In the clinical setting, oxygen supplementation between diagnosis and reattachment surgery may reduce the incidence and severity of glial-based complications, such as proliferative vitreoretinopathy. PMID- 10458172 TI - Age-related macular degeneration in grandparents of patients with Stargardt disease: genetic study. AB - PURPOSE: To report clinical features and molecular genetic study in three unrelated families in which age-related macular degeneration was observed in grandparents of patients with Stargardt disease. METHODS: A complete ophthalmologic examination including best-corrected visual acuity measurement, fundus examination, and fluorescein angiography was performed on all members of the three families. The entire coding sequence of the ABCR gene was analyzed using a combination of single strand conformation polymorphism and direct sequence analysis of the 50 exons. RESULTS: Compound heterozygous missense mutations were observed in patients with Stargardt disease (Arg212Cys, Argl107Cys, Gly1977Ser, Arg2107His, and le2113Met). Heterozygous missense mutations were observed in the grandparents with age-related macular degeneration (Arg212Cys and Arg1107Cys). CONCLUSIONS: We report phenotype and genotype findings in three unrelated families segregating patients with Stargardt disease and age-related macular degeneration. The hypothesis that the Arg212Cys and Arg1107Cys ABCR gene mutations could be susceptibility factors for age-related macular degeneration is discussed. We speculate that the relatives of patients affected with Stargardt disease who are carriers of heterozygous ABCR gene mutations may have a higher risk of developing age-related macular degeneration. PMID- 10458173 TI - Juvenile X-linked retinoschisis from XLRS1 Arg213Trp mutation with preservation of the electroretinogram scotopic b-wave. AB - PURPOSE: To present an Arg213Trp missense mutation in the XLRS1 gene in a family with juvenile X-linked retinoschisis in which one affected male had a normal electroretinogram scotopic b-wave amplitude. METHODS: Two affected males and one unaffected male from this family with X-linked retinoschisis underwent standard clinical examination including an electroretinogram. Mutations in the XLRS1 gene were detected by sequence analysis and by restriction enzyme assay for loss of an MSP-I restriction site. RESULTS: A missense mutation of C to T at nucleotide position 637 was identified in exon 6 of the XLRS1 gene. This changed the positively charged arginine to a nonpolar tryptophan (Arg213Trp) within the biologically important discoidin domain. Clinical examination revealed intraretinal cysts in a spoke-wheel distribution and early macular atrophy of the retinal pigment epithelium. Whereas the older affected patient had an "electronegative" electroretinogram typical of retinoschisis, the 13-year-old grandson with the same XLRS1 mutation had a normal electroretinogram scotopic b wave. CONCLUSION: Although the electroretinogram is a key diagnostic test for X linked retinoschisis, this report of a normal electroretinogram scotopic b-wave in a male with molecularly confirmed X-linked retinoschisis indicates that caution is advised in relying on the electroretinogram in differential diagnosis of this condition. PMID- 10458174 TI - Optical coherence tomography of cystoid macular edema associated with retinitis pigmentosa. AB - PURPOSE: To detect cystoid macular edema in consecutive eyes with retinitis pigmentosa by means of optical coherence tomography and to study the correlation between cross-sectional structures and angiographic findings in cystoid macular edema. METHODS: In a prospective study, cross-sectional images through the fovea were evaluated by means of optical coherence tomography in 89 phakic eyes of 46 patients with retinitis pigmentosa. Eyes showing cystoid appearance in the macula in the optical coherence tomographic images were further studied with measurement of the dimensions of cystoid lesions and with a fluorescein angiogram either at 18 minutes after dye injection or later. RESULTS: Cystoid lesions were observed in the macula in optical coherence tomographic images in 12 eyes in six (13%) of 46 patients. In these eyes, the width of total area of the cystoid lesions was positively correlated with the grade of fluorescein angiogram (Spearman rank correlation coefficient, r = .629; P = .029), but the thickness of the neurosensory retina at the center of the fovea was not. Among three variables for grading cystoid macular edema, consisting of angiographic grade, thickness of the neurosensory retina at the center of the fovea, and width of total area of the cystoid lesions, only the last measure was significantly correlated with best corrected visual acuity (Pearson correlation coefficient, r = .693; P = .012). CONCLUSION: Cystoid macular edema in eyes with retinitis pigmentosa could easily be detected with the use of optical coherence tomography independent of the angiographic degree of dye leakage. The size of cystoid lesions demonstrated in the optical coherence tomographic images, especially the thickness of the neurosensory retina at the center of the fovea, was not necessarily correlated with the angiographic grading of dye leakage. Measurement of the width of total area of the cystoid lesions in the optical coherence tomographic images is significantly correlated with the loss of visual acuity. PMID- 10458175 TI - High concentration of dexamethasone in aqueous and vitreous after subconjunctival injection. AB - PURPOSE: To determine the dexamethasone concentration in aqueous, vitreous, and serum of patients after a subconjunctival injection with dexamethasone disodium phosphate and to compare the effectiveness of a subconjunctival injection as a method of delivering dexamethasone into the vitreous with that of two previously tested routes: peribulbar injection and oral administration. METHODS: In a prospective study, 50 phakic patients who underwent a pars plana vitrectomy received a single subconjunctival injection with 2.5 mg of dexamethasone disodium phosphate, aqueous solution (after topical anesthesia and a subconjunctival injection with lidocaine) at varied intervals before surgery. An aqueous and a vitreous sample were taken from each patient, and serum samples were collected at multiple time points from nine of 50 patients. Dexamethasone concentrations were measured by radioimmunoassay. RESULTS: The estimated maximum dexamethasone concentration in the aqueous was 858 ng per ml at 2.5 hours after injection, and in the vitreous, 72.5 ng per ml at 3 hours. In serum, a mean maximum concentration of 32.4 ng per ml was measured at approximately 30 minutes after injection. CONCLUSIONS: Subconjunctival injection of 2.5 mg of dexamethasone disodium phosphate resulted in an estimated vitreous dexamethasone peak concentration three and 12 times higher, respectively, than after a peribulbar injection of 5 mg of dexamethasone disodium phosphate and an oral dose of 7.5 mg of dexamethasone. Thus, a subconjunctival injection is the most effective method of delivering dexamethasone into both the anterior and posterior segments of the eye. Systemic drug absorption is considerable and is of the same order of magnitude as after peribulbar injection. PMID- 10458176 TI - Follow-up of the original cohort with the Ahmed glaucoma valve implant. AB - PURPOSE: To study the long-term results of the Ahmed glaucoma valve implant in patients with complicated glaucoma in whom short-term results have been reported. METHODS: In this multicenter study, we analyzed the long-term outcome of a cohort of 60 eyes from 60 patients in whom the Ahmed glaucoma valve was implanted. Failure was characterized by at least one of the following: intraocular pressure greater than 21 mm Hg at both of the last two visits less than 6 mm Hg at both of the last two visits, loss of light perception, additional glaucoma surgery, devastating complications, and removal or replacement of the Ahmed glaucoma valve implant. Devastating complications included chronic hypotony, retinal detachment, malignant glaucoma, endophthalmitis, and phthisis bulbi; we also report results that add corneal complications (corneal decompensation or edema, corneal graft failure) as defining a devastating complication. RESULTS: The mean follow-up time for the 60 eyes was 30.5 months (range, 2.1 to 63.5). When corneal complications were included in the definition of failure, 26 eyes (43%) were considered failures. Cumulative probabilities of success at 1, 2, 3, and 4 years were 76%, 68%, 54%, and 45%, respectively. When corneal complications were excluded from the definition of failure, 13 eyes (21.5%) were considered failures. Cumulative probabilities of success at 1, 2, 3, and 4 years were 87%, 82%, 76%, and 76%, respectively. Most of the failures after 12 months of postoperative follow-up were because of corneal complications. CONCLUSIONS: The long-term performance of the Ahmed glaucoma valve implant is comparable to other drainage devices. More than 12 months after the implantation of the Ahmed glaucoma valve implant, the most frequent adverse outcome was corneal decompensation or corneal graft failure. These corneal problems may be secondary to the type of eyes that have drainage devices or to the drainage device itself. Further investigation is needed to identify the reasons that corneal problems follow drainage device implantation. PMID- 10458177 TI - Mooren ulcer in South India: serology and clinical risk factors. AB - PURPOSE: To investigate the rate of undiagnosed rheumatologic diseases and hepatitis C infection among patients with the clinical diagnosis of Mooren ulcer seen at Aravind Eye Hospital, Madurai, South India. METHODS: Twenty-one patients with the clinical diagnosis of Mooren ulcer and 44 control patients underwent a complete ophthalmic history and examination, as well as serologic testing for antinuclear antibodies, rheumatoid factor, antineutrophil cytoplasmic antibodies, herpes simplex virus 1 antibodies, and hepatitis C virus antibodies. RESULTS: There were no statistically significant differences in the rates of seropositivity for antinuclear antibodies, rheumatoid factor, antineutrophil cytoplasmic antibodies, herpes simplex virus 1 antibodies, and hepatitis C virus antibodies between patients with Mooren ulcer and control patients. Two patients with Mooren ulcer and four control patients were found to have a rheumatoid factor titer of greater than 1:20. One of the control patients, but none of the patients with Mooren ulcer, was found to have serologic evidence of hepatitis C infection. A history of corneal trauma, surgery, or infection was reported by 68% of patients with Mooren ulcer, compared with 20% of control patients (P < .001). Among patients with Mooren ulcer, bilateral disease occurred in 37% of patients, visual acuity was reduced to light perception in 15% of eyes, and perforation occurred in 19% of eyes. CONCLUSIONS: Nineteen (90%) of 21 patients with the clinical diagnosis of Mooren ulcer were found to have no evidence of an underlying rheumatologic disease by history, examination, or serologic testing, and none was seropositive for hepatitis C. However, patients with Mooren ulcer were more likely than control patients to report a history of corneal trauma, surgery, or infection. PMID- 10458178 TI - Efficacy of unilateral versus bilateral temporal artery biopsies for the diagnosis of giant cell arteritis. AB - PURPOSE: To determine the utility of unilateral versus bilateral temporal artery biopsies in detecting the pathologic changes of giant cell arteritis. METHODS: We performed a retrospective study to determine the utility of unilateral versus bilateral temporal artery biopsies in detecting the pathologic changes of giant cell arteritis. The pathologic reports of consecutive temporal artery biopsy specimens received at the Wilmer Ocular Pathology Laboratory over a 28-year period from 1968 to 1996 were reviewed. RESULTS: Of 908 specimens examined from 758 patients, 300 specimens were simultaneous bilateral biopsies from 150 patients, 72 specimens were bilateral sequential biopsies from 36 patients, and the remaining 536 specimens were unilateral biopsies from 536 patients. Of the 186 patients who had bilateral simultaneous or nonsimultaneous biopsies, 176 had identical diagnoses on both sides. In four patients, no artery was obtained on one side. In each of the remaining six patients, five of whom had bilateral simultaneous biopsies and one of whom had bilateral sequential biopsies performed 8 days apart, the biopsy specimen from one side was interpreted as showing only arteriosclerotic changes with no evidence of active or healed arteritis, whereas the other specimen was interpreted as showing either probable healed arteritis (three specimens) or possible early arteritis (three cases). In none of the six patients with differing diagnoses between the two sides was one side interpreted as showing definite, active giant cell arteritis. Five of the six patients were subsequently determined to have giant cell arteritis, based on a combination of clinical findings, erythrocyte sedimentation rate, and response to treatment with systemic corticosteroids. CONCLUSIONS: The results of this study indicate that performing a bilateral simultaneous or sequential temporal artery biopsy improves the diagnostic yield in at least 3% of cases of giant cell arteritis, whereas in 97% of cases, the two specimens show the same findings. Thus, in patients in whom only one artery can be biopsied, there is a high probability of obtaining the correct diagnosis. Nevertheless, although the improvement in diagnostic yield of bilateral temporal artery biopsies is low, the consequences of both delayed diagnosis and treatment of giant cell arteritis as well as the use of systemic corticosteroids in patients who do not have giant cell arteritis are of such potential severity that consideration should always be given to performing bilateral temporal artery biopsies in patients suspected of having the disease. PMID- 10458179 TI - Third cranial nerve palsy in children. AB - PURPOSE: To report the causes and the sensory, motor, and cosmetic results after treatment for oculomotor (third cranial nerve) palsy in children. METHODS: Review of the clinical records of children with a diagnosis of third cranial nerve palsy followed up in a university-based pediatric ophthalmology practice between 1981 and 1996. RESULTS: Forty-nine children with 53 affected eyes were followed up for a mean of 5.5 years. Third cranial nerve palsy was partial in 31 children (32 eyes) and complete in 18 children (21 eyes). The palsy was congenital in 20 eyes and caused by postnatal trauma in 17 eyes. Seventeen eyes had aberrant regeneration and four eyes with partial third cranial nerve palsy had spontaneous resolution. Thirty-six children (38 eyes) were affected before visual maturation (age 8 years), and 25 (27 eyes) had amblyopia. Of the five amblyopic eyes with quantifiable visual acuity, none had measurable improvement of Snellen visual acuity during the follow-up period. Overall, visual acuity was between 6/5 and 6/12 at the last follow-up visit in 31 eyes (58%). Ocular alignment was greatly improved after strabismus procedures, with a mean of 1.5 procedures for patients with partial third cranial nerve palsy and 2.3 procedures for those with complete palsy. Binocular function was difficult to preserve or restore but was achieved for some patients with partial third cranial nerve palsy. CONCLUSIONS: Surgical treatment of third cranial nerve palsy is frequently necessary, especially in cases of complete palsy. Multiple strabismus procedures are often needed to maintain good ocular alignment. Surgery can result in cosmetically acceptable alignment of the eyes, but it rarely results in restoration or achievement of measurable binocular function. Treatment of amblyopia is effective in maintaining the level of visual acuity present at the onset of the third cranial nerve palsy, but improvement in visual acuity is difficult to achieve. PMID- 10458180 TI - Assessment of central and peripheral fusion and near and distance stereoacuity in intermittent exotropic patients before and after strabismus surgery. AB - PURPOSE: To determine whether central fusion and distance stereoacuity are useful as objective measures in assessing the need for and success of surgery for intermittent exotropia (X[T]). METHODS: A prospective, institutional, clinical trial was conducted of 26 consecutive patients with X(T) who were undergoing strabismus surgery in whom fusion (central and peripheral) and stereoacuity (at near and distance) were assessed preoperatively and postoperatively, as well as in 112 normal subjects. To obtain accurate measurements with sensory tests, the lower age was limited to 5 years for inclusion. A successful surgical alignment was defined as an exotropia of 10 prism diopters or less at 6 m. Sensory and motor outcome measures were determined 1 year after surgery. RESULTS: The successful surgical alignment rate was 69%. All patients with X(T) demonstrated peripheral fusion, whereas 35% demonstrated central suppression preoperatively and postoperatively. Central fusion was not predictive of surgical outcome (P = .078); however, there was a trend toward less surgical success in patients with central suppression. Patients with X(T) exhibited good near stereoacuity before and after surgery. Distance stereoacuity in patients with X(T) preoperatively was significantly diminished compared with normal subjects (P < .001) and was improved in 58% postoperatively. Patients who achieved successful surgical alignment had a greater likelihood of demonstrating distance stereoacuity improvement postoperatively than patients who failed to achieve successful surgical alignment (P = .003). Patients with central suppression were unlikely to improve their distance stereoacuity postoperatively (P = .014). CONCLUSIONS: Successful surgery may improve distance stereoacuity. Better distance stereoacuity and central fusion are frequently associated with better surgical success in X(T). PMID- 10458181 TI - Does hyperoxygenation limit retinal degeneration after retinal detachment? PMID- 10458182 TI - Precrystalline posterior chamber intraocular lens for surgical correction of severe myopia. AB - PURPOSE: To report early experience with a posterior chamber precrystalline intraocular lens designed for correction of severe myopia from -10 to -30 diopters. This intraocular lens respects the transparent crystalline lens and does not interfere with accommodation. METHOD: The concave-convex methylmethacrylate lens, with flexible haptics that are supported in the sulcus, is placed behind the iris, in front of the lens, and centered with the pupil. RESULTS: From July 1995 to November 1998, 149 precrystalline lenses have been inserted with few, generally reversible complications, which are now largely prevented by adequate modification of the intraocular lens. Correction has been satisfactory and stable. CONCLUSION: Although follow-up is relatively short, precrystalline lens placement may be considered an important contribution to the surgical management of severe myopia to improve the patient's vision and, consequently, quality of life. PMID- 10458183 TI - Corneal endothelial changes in patients with chronic renal failure. AB - PURPOSE: To evaluate the morphologic changes of the corneal endothelium in patients with chronic renal failure. METHODS: Twenty corneas of 20 patients with chronic renal failure undergoing hemodialysis were examined by wide-field specular microscopy. Twenty normal corneas from 20 healthy age-matched subjects were enrolled as control corneas. RESULTS: Despite normal endothelial cell density, the corneal endothelium of patients with chronic renal failure showed marked polymegethism and pleomorphism. CONCLUSION: Polymegethism and pleomorphism of the corneal endothelium are found in patients with chronic renal failure. PMID- 10458184 TI - Noninfectious crystalline keratopathy after postoperative subconjunctival 5 fluorouracil. AB - PURPOSE: To describe a case of transient, noninfectious crystalline keratopathy after postoperative subconjunctival 5-fluorouracil. METHOD: Case report. Slit lamp biomicroscopic examinations were performed during serial office examinations. RESULTS: A 69-year-old woman underwent an uncomplicated trabeculectomy with adjunctive sponge 5-fluorouracil application for 5 minutes in her left eye. Postoperative treatment included topical prednisolone acetate, ciprofloxacin, and scopolamine. She was initially examined with intrastromal corneal crystalline deposits 5 days after her first postoperative subconjunctival injection of 5-fluorouracil. The deposits completely disappeared 4 days later without addition to or change in treatment. CONCLUSION: Transient, noninfectious, crystalline, intrastromal corneal deposits may develop after adjunctive subconjunctival 5-fluorouracil administration. PMID- 10458185 TI - Technique for harvesting keratolimbal allografts from corneoscleral buttons. AB - PURPOSE: To report a method for obtaining thin keratolimbal lenticules from cadaveric corneoscleral buttons for corneal epithelial stem cell allografts. METHOD: A standard silicone orbital sizing sphere and three 25-gauge needles were employed to fix the corneoscleral button to facilitate excision of keratolimbal lenticules. RESULT: Limbal stem cells in the form of keratolimbal lenticules were obtained without difficulty. CONCLUSION: With this technique, thin keratolimbal lenticules of good quality can be obtained efficiently, inexpensively, and without specialized instruments. PMID- 10458186 TI - The survival of herpes simplex virus in multidose office ophthalmic solutions. AB - PURPOSE: To determine the survival of herpes simplex virus type 1 (HSV-1) in several multidose ophthalmic solutions. METHODS: In three separate trials, 10 aliquots of 5 ml each of three common multidose topical ophthalmic solutions, sodium fluorescein, proparacaine, and nonpreserved artificial tears, were inoculated with 10(5), 10(4), and 10(3) pfu per ml of HSV-1. All samples were titered on A549 cells at various time points for surviving HSV-1. RESULTS: Herpes simplex virus type 1 was not recovered from the fluorescein and proparacaine solutions at 1 hour or any time thereafter, regardless of inoculation titer. Herpes simplex virus type 1 was recovered from the artificial tears up to 7 days. CONCLUSION: Unlike adenovirus, HSV-1 does not survive in preserved fluorescein and proparacaine multidose solutions; therefore, office transmission is highly unlikely. PMID- 10458187 TI - Microbiologic spectrum and susceptibility of isolates: part I. Postoperative endophthalmitis. Endophthalmitis Research Group. AB - PURPOSE: To present the microbial spectrum and susceptibilities of isolates in postoperative endophthalmitis. METHOD: Isolates from 206 eyes of 206 patients who underwent vitrectomy for postoperative endophthalmitis were examined. RESULTS: One-hundred twelve (54.4%) of 206 vitreous samples were culture positive and 14 (12.5%) of 112 culture-positive cases were polymicrobial, yielding a total of 126 isolates. Isolates included 59 (46.8%) gram-positive cocci, eight (6.3%) gram positive bacilli, 33 (26.2%) gram-negative organisms, five (4.0%) Actino-mycetes related organisms, and 21 (16.7%) fungi. Susceptibilities to amikacin, ceftazidime, chloramphenicol, cefazolin, ciprofloxacin, gentamicin, and vancomycin are reported. CONCLUSIONS: This is the largest, single-center, prospective series on microbial susceptibilities in postoperative endophthalmitis. We report a high prevalence of gram-negative species and fungi, suggesting that empiric therapy should include coverage for gram-negative pathogens and for fungal pathogens in appropriate settings. PMID- 10458188 TI - Microbiologic spectrum and susceptibility of isolates: part II. Posttraumatic endophthalmitis. Endophthalmitis Research Group. AB - PURPOSE: To present the microbial spectrum and susceptibilities of isolates in posttraumatic endophthalmitis. METHOD: Isolates from 182 eyes of 182 patients who underwent vitrectomy for posttraumatic endophthalmitis were examined. RESULTS: One hundred thirteen (62.1%) of 182 vitreous samples were culture-positive, and 23 (20.4%) of 113 culture-positive cases were polymicrobial, including three (2.7%) trimicrobial cases, yielding a total of 139 isolates. Isolates included 63 (45.3%) gram-positive cocci, 24 (17.3%) gram-positive bacilli, 25 (18.0%) gram negative organisms, seven (5.0%) Actinomycetes-related organisms, and 20 (14.4%) fungi. Susceptibilities to amikacin, ceftazidime, chloramphenicol, cefazolin, ciprofloxacin, gentamicin, and vancomycin are reported. CONCLUSIONS: This study represents a large series on microbial spectrum and susceptibilities in posttraumatic endophthalmitis. We report a high prevalence of gram-positive bacilli species and polymicrobial infections containing gram-negative species, underscoring the importance of broad-spectrum, combination antibiotics in the empiric treatment of posttraumatic endophthalmitis. PMID- 10458189 TI - Photocoagulation of iris nevus to control recurrent hyphema. AB - PURPOSE: To report a case of recurrent hyphema caused by an iris nevus that was successfully treated with photocoagulation. METHOD: Case report. In a 30-year-old woman with recurrent hyphema secondary to an iris nevus, photocoagulation was applied to the iris nevus. RESULTS: Before photocoagulation of the iris nevus, the patient had increasing frequency of hyphema episodes. After treatment, no hyphema episodes occurred during 13 months of observation. CONCLUSION: Photocoagulation of the surface of an iris nevus may prevent recurrent hyphema. PMID- 10458190 TI - Familial occurrence of retinitis punctata albescens and congenital sensorineural deafness. AB - PURPOSE: To report the cotransmission of retinitis punctata albescens (RPA) and congenital sensorineural deafness. METHODS: Case reports of two siblings with nyctalopia and profound bilateral sensorineural deafness. RESULTS: The affected siblings, an 11-year-old female and a 7-year-old male, presented with decreased visual acuity and night blindness. In both eyes of both siblings, ophthalmoscopic evaluation disclosed numerous white spots at the level of the retinal pigment epithelium with macular sparing. The rod threshold dark adaptation and electroretinogram tracings were consistent with advanced rod-cone degeneration. CONCLUSION: Two affected members of a family were found to exhibit RPA and congenital sensorineural deafness. This pedigree supports the genetic cotransmission of the traits. PMID- 10458191 TI - Idiopathic central retinal vein occlusion in a thrombophilic patient with the heterozygous 20210 G/A prothrombin genotype. AB - PURPOSE: To report the occurrence of monolateral central retinal vein occlusion in a patient with heterozygous 20210 G/A prothrombin genotype, known to be associated with high thrombophilic risk. METHODS: A monolateral central retinal vein occlusion was diagnosed in a 71-year-old woman, who had suffered from a deep vein thrombosis in her left leg at the age of 36 years. Mutations of the genes involved in the coagulation process were investigated by DNA polymerase chain reaction. RESULT: DNA analysis showed the patient to be heterozygous for the prothrombin 20210 G/A genetic variation. CONCLUSION: The 20210 G/A prothrombin gene mutation may be associated with central retinal vein occlusion. PMID- 10458192 TI - Photic maculopathy after pterygium excision. AB - PURPOSE: To report a patient who developed photic maculopathy after pterygium removal. METHODS: A 27-year-old woman underwent pterygium removal with a conjunctival autograft after administration of retrobulbar anesthesia. A coaxial operating microscope was used, with an estimated retinal exposure of approximately 40 minutes. RESULTS: On the first postoperative day, the patient noted a paracentral scotoma. A fluorescein angiogram on the fourth postoperative day documented a phototoxic lesion in the macula. CONCLUSIONS: Phototoxic injury to the macula may occur after pterygium removal. Ophthalmologists should take precautions to minimize prolonged intense coaxial illumination of the retina while performing any ocular microsurgery. PMID- 10458193 TI - Cytomegalovirus retinitis in HIV-infected patients with and without highly active antiretroviral therapy. AB - PURPOSE: To assess the impact of highly active antiretroviral therapy on the epidemiology of cytomegalovirus retinitis in patients infected with the human immunodeficiency virus (HIV). METHODS: In a study performed in a single center for infectious diseases, we compared the data collected in 1995 (without highly active antiretroviral therapy) with 1997 data (with highly active antiretroviral therapy). RESULTS: In a comparison of 1997 with 1995 data, the mean CD4+ cell count of patients with cytomegalovirus (CMV) retinitis was higher (169 +/- 150 CD4/microl vs 15 +/- 47 CD4/microl) (P = .05), and the relapses of CMV retinitis were less frequent (17% vs 36%) (P = .02). Newly diagnosed CMV retinitis decreased from 6.1% (59 of 952 patients) in 1995 to 1.2% (nine of 726 patients) in 1997 (P < .0001). In 1997, patients with newly diagnosed or relapsing CMV retinitis had a lower mean CD4+ (37 +/- 42) cell count than patients with no relapsing CMV retinitis (197 +/- 160) (P = .01). CONCLUSION: The incidence and recurrences of CMV retinitis decreased from 1995 to 1997, probably as a result of restored immunity while the patients were undergoing highly active antiretroviral therapy; however, the increasing frequency of HIV resistance to highly active antiretroviral therapy justifies close ocular follow-up. PMID- 10458194 TI - Choroidal melanoma with oculodermal melanocytosis in Hispanic patients. AB - PURPOSE: To describe three Hispanic patients with oculodermal melanocytosis and uveal melanoma. METHOD: Case series. RESULTS: Three Hispanic patients with oculodermal melanocytosis and uveal melanoma underwent enucleation. The diagnosis of choroidal melanoma was confirmed by histopathologic examination. CONCLUSION: In the Hispanic population, uveal melanoma can occur in the presence of oculodermal melanocytosis. PMID- 10458195 TI - Choroidal metastasis in men with metastatic breast cancer. AB - PURPOSE: To report two cases of choroidal metastasis in metastatic breast cancer in men. METHOD: Case reports of a 50-year-old man with an 8-year history of breast cancer who was initially examined with a solitary amelanotic choroidal tumor and a 62-year-old man with an 8-month history of breast cancer who was initially examined with numerous unilateral amelanotic choroidal tumors. RESULTS: Ophthalmoscopic and echographic characteristics of the choroidal tumors were typical for breast cancer metastasis. Systemic screening disclosed advanced metastatic disease in both patients. Choroidal metastasis could be effectively treated by external beam irradiation. CONCLUSIONS: Although breast cancer is a rare condition in men, it should be considered as a possible primary cancer in cases of choroidal metastasis. PMID- 10458196 TI - Apparent central nervous system depression in infants after the use of topical brimonidine. AB - PURPOSE: To report two cases in which topical brimonidine resulted in apparent central nervous system depression and unresponsiveness in an infant. METHODS: Review of two patients. An 11-day-old infant became lethargic and apneic after a single drop of brimonidine. These symptoms were reproduced after a second administration of brimonidine. A 5-month-old infant became lethargic and poorly responsive after receiving 1 drop of brimonidine in each eye. RESULTS: The first patient required admission to the hospital for medical stabilization. He recovered without sequelae. The second patient recovered spontaneously approximately 2.5 hours after administration of brimonidine. CONCLUSIONS: Topical brimonidine may be associated with central nervous system depression in infants. The use of brimonidine is not recommended in these patients until further data are available. PMID- 10458197 TI - Compression of the prechiasmatic optic nerve produces a junctional scotoma. AB - PURPOSE: To demonstrate the clinical-radiologic correlation between a junctional scotoma and a focal lesion compressing the prechiasmatic segment of the distal optic nerve. METHODS: Case report involving a man with a pituitary adenoma. Clinical correlation was determined by reviewing visual field evaluations and magnetic resonance images. RESULTS: The tumor compressed the prechiasmatic segment of the distal optic nerve but not the optic chiasm, producing a junctional scotoma documented by Goldmann and automated perimetry. The visual field defect resolved after neurosurgical decompression of the anterior visual pathway. CONCLUSIONS: A junctional scotoma can be caused by focal as well as large and diffuse lesions injuring the anterior visual pathway, specifically at the junction of the optic nerve and chiasm. This finding supports the existence of Wilbrand fibers. PMID- 10458198 TI - Corneal topographic evaluation of decentration in photorefractive keratectomy: treatment displacement vs intraoperative drift. PMID- 10458199 TI - Ultrasound biomicroscopy of ciliary body cysts. PMID- 10458200 TI - Refractive changes after pediatric intraocular lens implantation. PMID- 10458201 TI - Vitreopapillary traction as a cause of elevated optic nerve head. PMID- 10458202 TI - Docetaxel (Taxotere) in the treatment of non-small cell lung cancer: recent results and future developments. PMID- 10458203 TI - Overview of docetaxel (Taxotere) in the treatment of non-small cell lung cancer. AB - In four initial phase II studies of docetaxel administered as a 1-hour intravenous infusion at a dose of 100 mg/m2 every 3 weeks in chemotherapy-naive patients with advanced non-small cell lung cancer (NSCLC), the overall response rate was 31% in 128 evaluable patients and the median survival was 9 months. Five subsequent phase II studies of 100 mg/m2 docetaxel in the first-line setting showed similar results, with response rates in the range of 25% to 63% and favorable survival. Lower doses also have been evaluated in two trials using docetaxel 60 and 75 mg/m2, with overall response rates of 25% and 19%, respectively. In the second-line setting, after failure of first-line platinum based chemotherapy, four studies of docetaxel 100 mg/m2 have achieved response rates of 16% to 22% and encouraging median durations of survival of 30 to 42 weeks. In both first-and second-line settings, the toxicity of docetaxel is tolerable. Docetaxel is clearly an active drug in both the first- and second-line treatment of NSCLC, and recently conducted phase III trials will further define the role of this agent in the standard treatment of NSCLC. PMID- 10458204 TI - The use of docetaxel (Taxotere) in patients with advanced non-small cell lung cancer previously treated with platinum-containing chemotherapy regimens. AB - Several phase II studies have evaluated docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) at a dose of 100 mg/m2 administered as a 1-hour intravenous infusion every 3 weeks in the second-line treatment of non-small cell lung cancer (NSCLC). Two early phase II studies conducted at the University of Texas M.D. Anderson Cancer Center enrolled NSCLC patients who had failed prior platinum containing regimen, and response rates of 22% and 17%, respectively, were observed in evaluable patients. The estimated 1-year survival rate for both studies was 40%. In the later phase II studies, patients who had received prior NSCLC treatment were enrolled and response rates ranged from 15% to 22% in evaluable patients. A phase III randomized study was conducted based on these encouraging data. In this first randomized phase III trial of second-line chemotherapy for advanced NSCLC, patients who had been previously exposed to at least one course of platinum-containing chemotherapy received docetaxel at 100 mg/m2 or 75 mg/m2 versus a control regimen of vinorelbine or ifosfamide. This is the first phase III study that has compared the benefits of different chemotherapies in the second-line treatment of NSCLC. Data from this phase III study are forthcoming. These results may provide important objective indications of the quality of life benefits that can be achieved in patients with NSCLC in the second-line setting with an active agent such as docetaxel. PMID- 10458205 TI - Overview of docetaxel (Taxotere)/cisplatin combination in non-small cell lung cancer. AB - Cisplatin-based chemotherapy is effective in non-small cell lung cancer (NSCLC), although it prolongs survival only modestly. Single-agent docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) is highly active against NSCLC. The activity and tolerability of two docetaxel/ cisplatin regimens were therefore investigated in two multicenter phase II studies, one in Australia and one in France. Chemotherapy-naive patients with inoperable NSCLC received either docetaxel 75 mg/m2 on day 1 plus cisplatin 75 mg/m2 3 weekly (n = 47; Australian study) or docetaxel 75 mg/m2 on day 1 plus cisplatin 100 mg/m2 every 3 weeks for three cycles then every 6 weeks (n = 51; French study). The majority of the population (74%) had metastatic disease. Seventy-eight patients were evaluable for efficacy. Overall response rates were 36% (95% confidence interval, 25 to 47) in all evaluable patients and 34% in patients with metastases. Median duration of response was 6 months, with a 4-month median time to progression. Median survival time was 9 months, with a 1-year survival rate of 34%. A median of four (range, one to nine) treatment cycles were administered. Febrile neutropenia occurred in 14% of patients. Severe infection, which occurred in less than 7% of patients, led to two toxic deaths. Other severe toxicities were rare, with severe stomatitis and severe neurosensory side effects reported in 2% and 1%, respectively, of treated patients. No severe fluid retention occurred. Docetaxel/cisplatin, administered as two different schedules, is well tolerated and exhibits efficacy in the range of the most established combinations in the treatment of advanced NSCLC. PMID- 10458206 TI - Docetaxel (Taxotere) and gemcitabine in combination therapy. AB - In patients with advanced cancers, gemcitabine and docetaxel (Taxotere; Rhone Poulenc Rorer, Antony, France) can be administered together at 80% to 100% of their single-agent doses on days 1 and 8 of an every-3-week cycle without undue toxicity. In phase 1/11 studies, this combination has demonstrated activity against a range of tumors, including non-small cell lung cancer, and may represent an alternative to cisplatin-based chemotherapy. In advanced cancer, benefits in quality of life are likely to result. The fact that the combination of gemcitabine and docetaxel is generally well tolerated also suggests that the regimen should be assessed in the adjuvant and neoadjuvant settings. PMID- 10458207 TI - Docetaxel (Taxotere) in non-small cell lung cancer: ongoing studies in Heidelberg and future plans. AB - Docetaxel (Taxotere, Rhone-Poulenc Rorer, Antony, France) as a "stabilizing antitubulin" has in the last few years gained the reputation of being one of the most effective cytotoxic drugs in the therapy of non-small cell lung cancer. Clinical investigations of docetaxel in non-small cell lung cancer continue and currently focus on new combination chemotherapy regimens, its simultaneous use in combination with radiotherapy, as well as trials within multimodality treatment concepts. Our hospital's direct experiences with docetaxel include its sequential use with gemcitabine in single-agent therapy and a phase I study combining docetaxel with ifosfamide. In addition we are planning a randomized study to test docetaxel within a sequential and concomitant chemoradiotherapy concept in patients with inoperable stage III non-small cell lung cancer. PMID- 10458208 TI - Docetaxel (Taxotere) administered in weekly schedules. AB - The administration of a weekly low-dose taxane markedly reduces the severity of myelosuppression compared with a once-every-3-week schedule and allows the dose intensity (mg/m2/wk) of treatment to be increased. The dose-limiting toxicity observed in a weekly phase I trial was fatigue/asthenia. The maximum tolerated dose of a weekly docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) phase I study was 43 mg/m2; 36 mg/m2 was recommended for further study. This finding was similar to that in another phase I/II trial of weekly docetaxel in previously treated patients with metastatic breast cancer in which the recommended dose for the phase II study was 35 mg/m2. In this latter study, an objective response rate of 50% and a 0% incidence of febrile neutropenia have been reported. Other studies have been conducted to evaluate the efficacy and safety of the weekly schedule. One such study is an ongoing phase II trial in elderly or medically unfit patients with previously untreated advanced non-small cell lung cancer in whom weekly docetaxel appears active and well tolerated. Further investigation of weekly docetaxel alone or in combination is warranted. PMID- 10458209 TI - New approaches in the adjuvant and neoadjuvant therapy of non-small cell lung cancer, including docetaxel (Taxotere) combinations. AB - Among the issues debated in the therapy of early non-small cell lung cancer are whether postoperative chemotherapy improves survival, whether postoperative radiation therapy has some benefit either in local control or in the prevention of distant recurrence, and whether neoadjuvant treatment benefits patients with stage IIIA disease. The role of surgery is being investigated in the North American Intergroup Trial, in which concurrent chemoradiotherapy followed by surgery and postoperative chemotherapy is compared with concurrent chemoradiotherapy alone. Based on the evidence to date, surgery appears to benefit patients in whom complete resection is attained. However, even in these patients, the detection of tumor DNA in serum is a clear indication for postoperative chemotherapy. A trial undertaken by the Spanish Lung Cancer Group is currently investigating a novel neoadjuvant regimen involving gemcitabine, cisplatin, and weekly docetaxel (Taxotere; Rhone-Poulenc Rorer, Antony, France) in patients with mediastinoscopically confirmed N2 disease. PMID- 10458210 TI - Clinical radiosensitization: why it does and does not work. PMID- 10458211 TI - Radiosensitization with carboplatin for patients with unresectable stage III non small-cell lung cancer: a phase III trial of the Cancer and Leukemia Group B and the Eastern Cooperative Oncology Group. AB - PURPOSE: To determine whether the administration of carboplatin concurrently with radiation treatment improves survival in patients with inoperable stage III non small-cell lung cancer. PATIENTS AND METHODS: Two hundred eighty-three patients with inoperable stage III non-small-cell lung cancer were entered onto a randomized trial by the Cancer and Leukemia Group B and the Eastern Cooperative Oncology Group. Randomization was performed before initiation of any therapy. All patients received an induction chemotherapy program with vinblastine and cisplatin for 5 weeks, followed by 6,000 cGy of radiation therapy over 6 weeks. One hundred thirty-seven patients were randomized to this therapy regimen alone; 146 patients were randomized to receive carboplatin at 100 mg/m2/wk concurrent with the radiation therapy. RESULTS: The complete response was 18% with concurrent carboplatin versus 10% with radiotherapy alone (P = .101). There was no difference with respect to failure-free survival (10% with carboplatin and 9% with radiotherapy alone) or overall survival (13% with carboplatin and 10% with radiotherapy alone) at 4 years. In patients not receiving carboplatin, the relapse rate was 69% within the field of radiation and 53% in the boost volume. In patients receiving carboplatin, the relapse rate was 59% within the field of radiation and 43% in the boost volume. Patients with cancers more than 70 cm2 in size had significantly poorer survival (P = .01). CONCLUSION: Carboplatin at the dose and schedule used did not significantly impact on disease control or survival. The relapse rate within the chest remained more than 50%. More effective regimens will be required to impact on local disease control and survival. PMID- 10458212 TI - Randomized phase III study of gemcitabine-cisplatin versus etoposide-cisplatin in the treatment of locally advanced or metastatic non-small-cell lung cancer. AB - PURPOSE: We conducted a randomized trial to compare gemcitabine-cisplatin with etoposide-cisplatin in the treatment of patients with advanced non-small-cell lung cancer (NSCLC). The primary end point of the comparison was response rate. PATIENTS AND METHODS: A total of 135 chemotherapy-naive patients with advanced NSCLC were randomized to receive either gemcitabine 1,250 mg/m2 intravenously (IV) days 1 and 8 or etoposide 100 mg/m2 IV days 1 to 3 along with cisplatin 100 mg/m2 IV day 1. Both treatments were administered in 21-day cycles. One hundred thirty-three patients were included in the intent-to-treat analysis of response. RESULTS: The response rate (externally validated) for patients given gemcitabine cisplatin was superior to that for patients given etoposide-cisplatin (40.6% v 21.9%; P = .02). This superior response rate was associated with a significant delay in time to disease progression (6.9 months v 4.3 months; P = .01) without an impairment in quality of life (QOL). There was no statistically significant difference in survival time between both arms (8.7 months for gemcitabine cisplatin v 7.2 months for etoposide-cisplatin; P = .18). The overall toxicity profile for both combinations of drugs was similar. Nausea and vomiting were reported more frequently in the gemcitabine arm than in the etoposide arm. However, the difference was not significant. Gemcitabine-cisplatin produced less grade 3 alopecia (13% v 51%) and less grade 4 neutropenia (28% v 56% ) but more grade 3 and 4 thrombocytopenia (56% v 13%) than did etoposide-cisplatin. However, there were no thrombocytopenia-related complications in the gemcitabine arm. CONCLUSION: Compared with etoposide-cisplatin, gemcitabine-cisplatin provides a significantly higher response rate and a delay in disease progression without impairing QOL in patients with advanced NSCLC. PMID- 10458213 TI - Disseminated tumor cells in lymph nodes as a determinant for survival in surgically resected non-small-cell lung cancer. AB - PURPOSE: In recent years, the detection of even a few tumor cells in lymph nodes of patients with surgically resected non-small-cell lung cancer (NSCLC) became possible with immunohistochemical staining procedures. Tumor cells in lymph nodes have been shown to be associated with an increased rate of early recurrence. However, the prognostic significance of this minimal tumor cell spread for overall survival remains unclear. PATIENTS AND METHODS: We used the epithelium specific monoclonal antibody Ber-EP4, which recognizes the 17-1A antigen (also called EGP40 or Ep-CAM), to discover small tumor cell deposits (< or = three cells) in 565 regional lymph nodes judged as tumor-free by conventional histopathology in patients with NSCLC staged as pT1-4, pN0-2, M0, R0. In a prospective analysis, we studied the influence of the detected tumor cells on the cancer recurrence rate and survival of 117 patients. RESULTS: Ber-EP4-positive cells were found in 27 of 125 patients (21.6%). After an observation period of 64 months, patients with disseminated tumor cells had reduced disease-free survival (P < .0001) and overall survival (P = .0001) rates in univariate analyses (logrank test). Multivariate analysis (Cox model) showed a 2.7 times increased risk for tumor relapse and a 2.5 times increased risk for shorter survival in patients with disseminated tumor cells compared with patients without such cells. Patients without any evidence of histopathologic and immunohistochemical lymph node involvement had an overall survival rate of 78%. CONCLUSION: The immunohistochemical detection of disseminated tumor cells in lymph nodes of patients with completely resected NSCLC is an independent prognostic factor for overall survival. PMID- 10458214 TI - Cisplatin and gemcitabine treatment for malignant mesothelioma: a phase II study. AB - PURPOSE: We performed a phase II study of combined cisplatin 100 mg/m2, given intravenously on day 1, and gemcitabine 1,000 mg/m2, given intravenously on days 1, 8, and 15 of a 28-day cycle for six cycles among patients with advanced measurable pleural mesothelioma. PATIENTS AND METHODS: Pleural tumor was measured at three levels on computed tomographic scans at study entry and before the second, fourth, and sixth cycles and every 2 months thereafter to disease progression. Of the 21 patients treated, 19 were male; the median age was 62 years (range, 46 to 74 years); 62% had epithelial tumors; and 18 were classified as tumor-node-metastasis system stage III or IV. Ninety-four cycles were given (median, six; mean, 4.5 per patient), with a mean relative dose intensity of cisplatin 96.7% and gemcitabine 82.5%. RESULTS: Best objective responses achieved were as follows: complete response, no patients; partial response, 10 patients (complete response + partial response, 47.6% [95% confidence interval, 26.2% to 69.0%]); no change, nine patients; and progressive disease, two patients. Median response duration was 25 weeks, progression-free survival was 25 weeks, and overall survival was 41 weeks. Nine of the 10 responders (90%) and three of nine patients with no change had significant symptom improvement. Serial measurements of vital capacity were performed on three of the responders; all showed a significant increase during the time of remission. Toxicity was mainly gastroenterologic and hematologic. Grade 3 nausea and vomiting occurred in 33% of patients, grade 3 leukopenia in 38%, grade 3 thrombocytopenia in 14%, and grade 4 thrombocytopenia in 19%. CONCLUSION: Combined cisplatin and gemcitabine is an active combination in malignant mesothelioma and produces symptomatic benefit in responding patients. PMID- 10458215 TI - Bromodeoxyuridine alternating with radiation for advanced uterine cervix cancer: a phase I and drug incorporation study. AB - PURPOSE: Preclinical studies show a significant increase in the ratio of the radiosensitizer bromodeoxyuridine (BUdR) in tumors versus the intestinal mucosa during the drug elimination period, compared with the ratio during drug infusion. We constructed a phase I study in patients with locally advanced cervix cancer, using alternating cycles of BUdR and radiation therapy (RT). PATIENTS AND METHODS: Eighteen patients with stage IIB to IVA cervix cancer participated. A treatment cycle consisted of a 4-day BUdR infusion followed by a week of pelvic RT, 15 Gy twice daily in 1.5-Gy fractions. After three cycles, additional BUdR was infused, followed by brachytherapy. The fraction of thymidine replaced by BUdR and the fraction of cells incorporating BUdR were determined in rectal mucosa and tumor biopsies at the end of the first BUdR infusion (day 5), at the middle of the first RT week (day 10), and at the time of brachytherapy. RESULTS: Dose-limiting toxicity was observed in one of 16 patients receiving 1,000 mg/m2/d x 4 days and in both patients receiving 1,333 mg/m2/d x 4 days each cycle. After a median follow-up of 39 months, 12 patients (66%) were free of pelvic disease and nine (50%) were alive and disease free. The ratio of tumor to rectum BUdR incorporation averaged 1.5 to 1.8 and did not differ significantly between day 5 and day 10. A trend toward reduced ratio was observed at brachytherapy. Drug containing cells in rectal biopsies migrated from the crypts to the mucosal surface. CONCLUSION: In this schedule, 1,000 mg/m2/d is the maximum-tolerated dose of BUdR. BUdR incorporation levels in tumors were consistent with clinically significant radiosensitization. The migration of BUdR-containing rectal mucosa cells from the crypts to the surface at the time of RT suggests that this regimen may offer a relative sparing of the mucosa from radiosensitization. PMID- 10458216 TI - Positron emission tomography for evaluating para-aortic nodal metastasis in locally advanced cervical cancer before surgical staging: a surgicopathologic study. AB - PURPOSE: Positron emission tomographic (PET) scanning provides a novel means of imaging malignancies. This prospective study was undertaken to evaluate PET scanning in detecting para-aortic nodal metastasis in patients with locally advanced cervical carcinoma and no evidence of extrapelvic disease before planned surgical staging lymphadenectomy. MATERIALS AND METHODS: After 20 mCi of 2 [18F]fluoro-2-deoxy-D-glucose (FDG) were administered intravenously, the abdomen and pelvis were scanned. Continuous bladder irrigation was used to reduce artifact. Patients were classified by the presence or absence of FDG uptake in the primary tumor and in pelvic or para-aortic nodes. Para-aortic node metastases were classified as present or absent according to a standardized staging procedure. Pelvic node metastases were similarly classified in a subset of patients who underwent pelvic node resection. RESULTS: Thirty-two patients with stage IIB (n = 6), IIIB (n = 24), and IVA (n = 2) tumors were studied. Fluorodeoxyglucose was taken up by 91% of the cervical tumors. Six of eight patients with positive para-aortic node metastasis had PET scan evidence of para aortic nodal metastasis. One of the two false-negatives had only one microscopic focus of metastatic cancer. In the para-aortic nodes, PET scanning had a sensitivity of 75%, a specificity of 92%, a positive predictive value of 75%, and a negative predictive value of 92%. Fluorodeoxyglucose para-aortic nodal uptake conferred a relative risk of 9.0 (95% confidence interval, 2.3 to 36.0) for para aortic nodal metastasis. All 10 of 17 patients with metastasis were predicted by PET scanning (P < .001); five of these patients had abnormalities on computed tomographic scans. CONCLUSION: Cervical cancers have a high avidity for FDG. The use of PET-FDG scanning accurately predicts both the presence and absence of pelvic and para-aortic nodal metastatic disease. PMID- 10458217 TI - First-line chemotherapy with epirubicin, paclitaxel, and carboplatin for advanced ovarian cancer: a phase I/II study of the Arbeitsgemeinschaft Gynakologische Onkologie Ovarian Cancer Study Group. AB - PURPOSE: Despite the progress that has been achieved over the years, survival rates in patients with advanced ovarian cancer are still disappointing. New methods to improve the efficiency of first-line chemotherapy are warranted. One method to improve results is to add more non-cross-resistant drugs to platinum paclitaxel combination regimens. Anthracyclines are among the candidates for incorporation as the "third drug" into first-line regimens for advanced ovarian cancer. PATIENTS AND METHODS: We performed a phase I/II trial with escalating doses of epirubicin (60, 75, and 90 mg/m2) combined with fixed doses of paclitaxel and carboplatin in 27 previously untreated patients with advanced gynecologic malignancies. RESULTS: Dose-limiting toxicity occurred at dose level 2 (75 mg/m2 epirubicin) and consisted of myelosuppression (neutropenia, thrombocytopenia). No dose-limiting, nonhematologic toxicities were observed. The maximum tolerable dose was epirubicin 60 mg/m2 (E) combined with a 3-hour infusion of paclitaxel 175 mg/m2 (T) and carboplatin AUC 5 (Carbo). Preliminary analysis indicated promising activity against ovarian cancer. CONCLUSION: The three-drug combination ET-Carbo, given according to the outlined dose and schedule, should be considered for further phase III evaluation. A randomized German-French intergroup trial comparing ET-Carbo with carboplatin-paclitaxel has already been initiated. PMID- 10458218 TI - Randomized phase III trial comparing the new potent and selective third generation aromatase inhibitor vorozole with megestrol acetate in postmenopausal advanced breast cancer patients. North American Vorozole Study Group. AB - PURPOSE: To compare the efficacy and safety of vorozole (VOR) 2.5 mg once daily with that of megestrol acetate (MA) 40 mg four times per day as second-line therapy in postmenopausal women with advanced breast cancer whose disease progressed after tamoxifen treatment. PATIENTS AND METHODS: A total of 452 patients were enrolled onto an open, multicenter, randomized phase III trial comparing VOR to MA for tumor response, safety, and quality of life (as indicated by the Functional Living Index-Cancer score). RESULTS: Vorozole produced a response rate of 9.7%, compared with 6.8% for MA (P = .24). Clinical benefit (complete response + partial response + no change in > 6 months) was demonstrated in 23.5% and 27.2% of patients treated with VOR and MA, respectively (P = .42). Median duration of response was 18.2 months for VOR versus 12.5 months for MA (P = .074). There was no significant difference in time to progression or survival between the treatment groups. Discontinuation of treatment because of adverse events occurred less frequently in the VOR-treated group (3.1% v 6.2%; P = .18). Patients on the VOR arm reported significantly more nausea, hot flushes, arthralgia, upper respiratory tract infection, anorexia, and paresthesia, whereas those treated with MA had significantly more dyspnea, increased appetite, and weight increase. There was no difference between the two treatment groups in Functional Living Index-Cancer scores (total or subscales). However, when analyzed by objective response, patients with complete or partial responses (P = .032) or no change (P = .033) who were receiving VOR had significant improvement in the psychologic well-being subscale, compared with patients given MA. CONCLUSION: Vorozole is well tolerated and as effective as MA in the treatment of postmenopausal advanced breast cancer patients with disease progression after tamoxifen treatment. PMID- 10458219 TI - Dose-response trial of megestrol acetate in advanced breast cancer: cancer and leukemia group B phase III study 8741. AB - PURPOSE: To investigate whether dose escalation of megestrol acetate (MA) improves response rate and survival in comparison with standard doses of MA. PATIENTS AND METHODS: Three hundred sixty-eight patients with metastatic breast cancer, positive and/or unknown estrogen and progesterone receptors, zero or one prior trial of hormonal therapy, and no prior chemotherapy for metastatic disease were prospectively randomized into three groups. The groups of patients received either MA 160 mg/d (one tablet per day), MA 800 mg/d (five tablets per day), or MA 1,600 mg/d (10 tablets per day). RESULTS: Patient characteristics were well balanced in the three treatment groups. Three hundred sixty-six patients received treatment and were included in the analyses. The response rates were 23%, 27%, and 27% for the 160-mg, 800-mg, and 1,600-mg arms, respectively. Response duration correlated inversely with dose. Median durations of response were 17 months, 14 months, and 8 months for the 160-mg, 800-mg, and 1,600-mg arms, respectively. No significant differences in the treatment arms were noted for time to disease progression or for survival; survival medians were 28 months (low dose), 24 months (mid dose) and 29 months (high dose). The most frequent and troublesome toxicity, weight gain, was dose-related, with approximately 20% of patients on the two higher-dose arms reporting weight gain of more than 20% of their prestudy weight, compared with only 2% in the 160-mg dose arm. CONCLUSION: With a median follow-up of 8 years, these results demonstrate no advantage for dose escalation of MA in the treatment of metastatic breast cancer. PMID- 10458220 TI - Vinorelbine and paclitaxel as first-line chemotherapy in metastatic breast cancer. AB - PURPOSE: To evaluate the efficacy and toxicity of a combination of vinorelbine (VNB) and paclitaxel (PTX) as first-line chemotherapy in metastatic breast carcinoma (MBC). PATIENTS AND METHODS: Between August 1995 and August 1997, 49 patients with untreated MBC received a regimen that consisted of VNB 30 mg/m2 in a 20-minute intravenous (IV) infusion on days 1 and 8 and PTX 135 mg/m2 in a 3 hour IV infusion (starting 1 hour after VNB) on day 1. Cycles were repeated every 28 days. The median age of the patients was 52 years, and 59% of patients were postmenopausal. Median performance status was 1. Dominant sites of disease were soft tissue in 6%, bone in 29%, and viscera in 65%. RESULTS: Objective responses were recorded in 27 of 45 assessable patients (60%; 95% confidence interval, 46% to 74%). Complete remissions occurred in three patients (7%), and partial remissions occurred in 24 patients (53%). No change was recorded in 12 patients (27%), and progressive disease occurred in six patients (13%). The median time to treatment failure was 7 months, and median survival duration was 17 months. The limiting toxicity was myelosuppression, mainly leukopenia in 49 patients (100%) (grade 1 to grade 2, four patients; grade 3, 30 patients; and grade 4, 15 patients). Neutropenia was observed in 100% of patients (grade 1 to grade 2, three patients; grade 3, 11 patients; grade 4, 35 patients). Two treatment related deaths due to febrile neutropenia were observed in patients with massive liver involvement. Peripheral neurotoxicity developed in 33 patients (67%) (grade 1, 25 patients; grade 2, eight patients); there were no grade 3 or grade 4 episodes. CONCLUSION: The combination of VNB-PTX showed significant activity as first-line chemotherapy for patients with MBC. Myelosuppression was the dose limiting side effect, whereas neurotoxicity was mild to moderate. PMID- 10458221 TI - Multicycle high-dose chemotherapy and filgrastim-mobilized peripheral-blood progenitor cells in women with high-risk stage II or III breast cancer: five-year follow-up. AB - PURPOSE: To determine the safety and efficacy of multiple cycles of dose intensive, nonablative chemotherapy in women with poor-prognosis breast cancer. PATIENTS AND METHODS: Women with stage II breast cancer and 10 or more involved nodes or four or more involved nodes and estrogen receptor-negative tumors and women with stage III disease received three cycles of epirubicin 200 mg/m2 and cyclophosphamide 4 g/m2, with progenitor cell and filgrastim support every 28 days (n = 79) or 21 days (n = 20). Patients were reviewed at least twice yearly thereafter. Twenty-six patients had bone marrow and apheresis collections assessed for the presence of micrometastatic tumor cells. RESULTS: Ninety-nine women (median age, 43 years; range, 24 to 60 years) were treated. Ninety-two completed all three cycles of chemotherapy. The major toxicity was severe, reversible myelosuppression that was more prolonged with successive cycles, and this did not differ between patients given treatment every 28 days and those treated every 21 days. Febrile neutropenia occurred in 176 (61%) of 287 cycles. Severe mucositis (grade 3 or 4) occurred in 23% of cycles but tended to be short lived and was reversible. The cardiac ejection fraction fell by a median of 4% during treatment, and three patients developed evidence of cardiac failure after chemotherapy. Two patients (2%) died of acute toxicity. Three of 26 patients had evidence of circulating micrometastatic tumor cells. The actuarial distant disease-free and overall survival rates at 60-month follow-up were 64% (95% confidence interval [CI], 53% to 75%) and 67% (95% CI, 56% to 78%), respectively. CONCLUSION: Multiple cycles of dose-intensive, nonablative chemotherapy is a feasible and safe approach. Disease control and survival are similar to those in other studies of myeloablative chemotherapy in poor-prognosis breast cancer. The regimen is being evaluated in a randomized trial of the International Breast Cancer Study Group. PMID- 10458222 TI - Sequential dose-dense doxorubicin, paclitaxel, and cyclophosphamide for resectable high-risk breast cancer: feasibility and efficacy. AB - PURPOSE: Dose-dense chemotherapy is predicted to be a superior treatment plan. Therefore, we studied dose-dense doxorubicin, paclitaxel, and cyclophosphamide (A ->T-->C) as adjuvant therapy. METHODS: Patients with resected breast cancer involving four or more ipsilateral axillary lymph nodes were treated with nine cycles of chemotherapy, using 14-day intertreatment intervals. Doses were as follows: doxorubicin 90 mg/m2 x 3, then paclitaxel 250 mg/m2/24 hours x 3, and then cyclophosphamide 3.0 g/m2 x 3; all doses were given with subcutaneous injections of 5 microg/kg granulocyte colony-stimulating factor on days 3 through 10. Amenorrheic patients with hormone receptor-positive tumors received tamoxifen 20 mg/day for 5 years. Patients treated with breast conservation, those with 10 or more positive nodes, and those with tumors larger than 5 cm received radiotherapy. RESULTS: Between March 1993 and June 1994, we enrolled 42 patients. The median age was 46 years (range, 29 to 63 years), the median number of positive lymph nodes was eight (range, four to 25), and the median tumor size was 3.0 cm (range, 0 to 11.0 cm). The median intertreatment interval was 14 days (range, 13 to 36 days), and the median delivered dose-intensity exceeded 92% of the planned dose-intensity for all three drugs. Hospital admission was required for 29 patients (69%), and 28 patients (67%) required blood product transfusion. No treatment-related deaths or cardiac toxicities occurred. Doxorubicin was dose reduced in four patients (10%) and paclitaxel was reduced in eight (20%). At a median follow-up from surgery of 48 months (range, 3 to 57 months), nine patients (19%) had relapsed, the actuarial disease-free survival rate was 78% (95% confidence interval, 66% to 92%), and four patients (10%) had died of metastatic disease. CONCLUSION: Dose-dense sequential adjuvant chemotherapy with doxorubicin, paclitaxel, and cyclophosphamide (A-->T-->C) is feasible and promising. Several ongoing phase III trials are evaluating this approach. PMID- 10458223 TI - Local failure is responsible for the decrease in survival for patients with breast cancer treated with conservative surgery and postoperative radiotherapy. AB - PURPOSE: The aim of the present study was to evaluate the role of local failure (LF) in the survival of patients treated with lumpectomy and postoperative radiotherapy and to investigate whether LF is not only a marker for distant metastasis (DM) but also a cause. METHODS: Charts of patients treated with breast conservative surgery between 1969 and 1991 were reviewed retrospectively. There were 2,030 patients available for analysis. The median duration of follow-up was 6 years. A Cox regression multivariate analysis was performed using LF as a time dependent covariate. RESULTS: Local control (LC) was 87% at 10 years. Local failure led to poorer survival at 10 years than local control (55% v 75%, P < .00). In a Cox model, local failure was a powerful predictor of mortality. The relative risk associated with LF was 3.6 for mortality and 5.1 for DM (P < .00). In patients with LF, the rate of DM peaked at 5 to 6 years, whereas it peaked at 2 years for patients with LC. The mean time between surgery and DM was 1,050 days for patients without LF and 1,650 days for patients with LF (P < .00). CONCLUSION: Our results show that local failure is associated with an increase in mortality. The difference in the time distribution of distant metastasis for LF and LC could imply distinct mechanisms of dissemination. Local failure should be considered not only as a marker of occult circulating distant metastases but also as a source for new distant metastases and subsequent mortality. PMID- 10458224 TI - Utility of magnetic resonance imaging in the management of breast cancer: evidence for improved preoperative staging. AB - PURPOSE: The staging and treatment for breast cancer are changing; there is an increase in the incidence of ductal carcinoma-in-situ, the use of fine-needle aspiration and stereotactic biopsy for diagnosis, and the use of neoadjuvant chemotherapy. Thus, there is a need for a tool to assess more precisely the extent of cancer in the breast before surgery. To better plan surgical and chemotherapeutic interventions, we evaluated high-resolution magnetic resonance imaging (MRI) as such a tool. PATIENTS AND METHODS: Fifty-seven patients with 58 cases of breast cancer were evaluated preoperatively with MRI using a technique called the triple-acquisition rapid gradient echo technique to maximize anatomic detail. Imaging results were compared with mammography and subsequent pathology results. RESULTS: Magnetic resonance imaging correctly identified residual or primary cancer in 55 of 58 cases and accurately predicted the extent of the cancer in 54 of 58 cases. The anatomic extent was more accurately defined with MRI compared with mammography (98% v 55%). Magnetic resonance imaging added the greatest value in cases of multifocal disease. CONCLUSION: By applying MRI selectively to patients with a known diagnosis of cancer and focusing on defining the extent of malignant lesions, we were able to obtain clear and accurate anatomic information. Our results suggest that MRI could provide very valuable information for preoperative planning and single-stage resection in breast cancer. Based on preliminary data from our series, MRI would be valuable as a staging tool in the preoperative setting even if the cost is in the range of $1,300 to $2,000. It is already significantly less than the target cost, so it is reasonable to refine this technique for clinical use to help plan the most appropriate surgical intervention and possibly reduce costs as well. A careful prospective study is warranted to validate our findings. PMID- 10458225 TI - Adjuvant treatment and onset of menopause predict weight gain after breast cancer diagnosis. AB - PURPOSE: Weight gain is common during the first year after breast cancer diagnosis. In this study, we examined clinical factors associated with body size at diagnosis and weight gain during the subsequent year. PATIENTS AND METHODS: An inception cohort of 535 women with newly diagnosed locoregional breast cancer underwent anthropometric measurements at baseline and 1 year. Information was collected on tumor- and treatment-related variables, as well as diet and physical activity. RESULTS: Mean age was 50.3 years; 57% of women were premenopausal. Mean baseline body mass index (weight [kg] divided by height [m] squared) was 25.5 kg/m2. Overall, 84.1% of the patients gained weight. Mean weight gain was 1.6 kg (95% confidence interval, 1.2 to 1.9 kg), 2.5 kg (95% confidence interval, 1.8 to 3.2 kg) in those receiving chemotherapy, 1.3 kg (95% confidence interval, 0.7 to 1.8 kg) in those receiving tamoxifen only, and 0.6 kg (95% confidence interval, 0.01 to 1.3 kg) in those receiving no adjuvant treatment. Menopausal status at diagnosis (P = .02), change in menopausal status over the subsequent year (P = .002), axillary nodal status (P = .009), and adjuvant treatment (P = .0002) predicted weight gain in univariate analysis. In multivariate analysis, onset of menopause and administration of chemotherapy were independent predictors of weight gain (all P < or = .05). Caloric intake decreased (P < .01) and physical activity increased (P < .05) during the year after diagnosis; these factors did not explain the observed weight gain. CONCLUSION: Weight gain is common after breast cancer diagnosis; use of adjuvant chemotherapy and onset of menopause are the strongest clinical predictors of this weight gain. PMID- 10458226 TI - Elements of informed consent for hormone replacement therapy in patients with diagnosed breast cancer. AB - PURPOSE: An approach to providing informed consent to breast cancer survivors considering hormone replacement therapy (HRT) is offered. METHODS: Current information on HRT, breast cancer, and chronic disease prevention is reviewed in the context of risks faced by women with resected breast cancer. RESULTS: Breast cancer patients, unwilling to trade symptom reduction for even a small increase in recurrence risk, are at substantially increased risk of death from breast cancer relative to other causes. Observational studies suggest that long-term HRT increases breast cancer development. The influence of HRT on the growth of established breast cancer has not been determined; however, estrogen reduction (oophorectomy) significantly reduces recurrence in premenopausal women, and current evidence cannot exclude a risk that HRT increases recurrence to the same degree. The following issues are of particular relevance to breast cancer survivors: HRT reduces mammographic sensitivity, increases thromboembolic events, and increases endometrial cancer risk. Although benefit for HRT is commonly inferred from observational studies, randomized trials of HRT on all-cause mortality have not been completed. For coronary heart disease prevention, an array of strategies independent of HRT are available, with some (tamoxifen, selective estrogen receptor modifiers [SERMs], diet, and exercise) likely to favorably influence breast cancer risk; for osteoporosis prevention, an array of strategies also are available, with some (bisphosphonates, tamoxifen, SERMs, and exercise) likely to favorably influence breast cancer risk. CONCLUSION: Current data preclude the generation of evidence-based guidelines for HRT use in breast cancer survivors, and clinical trials in this setting should be supported. However, given available therapeutic alternatives for menopausal symptom management and chronic disease prevention, breast cancer survivors should be offered HRT only with caution and with their full participation in the decision making process. PMID- 10458227 TI - Physical and psychological morbidity after axillary lymph node dissection for breast cancer. AB - PURPOSE: Alternatives to axillary lymph node dissection (ALND) are being developed amid controversy surrounding the therapeutic benefit and overall utility of this routine surgical procedure. Although potential negative side effects associated with ALND are known, we set out to examine whether these side effects contribute significantly to patient reports of quality of life and mental health. PATIENTS AND METHODS: We surveyed 222 women who had received an ALND as part of breast cancer surgery. All women underwent a physical therapy assessment of range of arm/shoulder motion and completed the Modified Post-operative Pain Questionnaire, the Pain Disability Index, the McGill Pain Questionnaire (short form), the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire, and the Mental Health Inventory. RESULTS: Seventy-two percent of the women experienced arm/shoulder pain, weakness, or numbness in the week before the interview, and range of motion of the affected arm/shoulder was impaired in 73% of the women. Severity of pain was reported to be low to moderate, and younger patients experienced greater pain than older patients. Pain severity correlated positively with the number of lymph nodes removed and receipt of chemotherapy and was not significantly related to length of time since surgery or receipt of radiation therapy. Generally high levels of cancer-specific quality of life and mental health were reported. Quality of life was significantly predicted by the McGill Pain Questionnaire, and mental health was significantly predicted by the Pain Disability Index and the physical therapy assessment. CONCLUSION: Surgery-related symptoms after ALND persist for a majority of women with breast cancer and are not significantly related to time since surgery or receipt of radiation therapy. These symptoms and associated disability are significantly predictive of cancer-specific quality of life and mental health. PMID- 10458228 TI - Prognostic factors for the outcome of chemotherapy in advanced soft tissue sarcoma: an analysis of 2,185 patients treated with anthracycline-containing first-line regimens--a European Organization for Research and Treatment of Cancer Soft Tissue and Bone Sarcoma Group Study. AB - PURPOSE: A total of 2,185 patients with advanced soft tissue sarcomas who had been treated in seven clinical trials investigating the use of doxorubicin- or epirubicin-containing regimens as first-line chemotherapy were studied in this prognostic-factor analysis. PATIENTS AND METHODS: Overall survival time (median, 51 weeks) and response to chemotherapy (26% complete response or partial response) were the two end points. The cofactors were sex; age; performance status; prior therapies; the presence of locoregional or recurrent disease; lung, liver, and bone metastases at the time of entry onto the trial; long time period between the initial diagnosis of sarcoma and entry onto the study; and histologic type and grade. RESULTS: Univariate analyses showed (a) a significant, favorable influence of good performance status, young age, and absence of liver metastases on both survival time and response rate, (b) a significant, favorable influence of low histopathologic disease grade on survival time, despite a significantly lower response rate, (c) increased survival time for patients with a long time period between the initial diagnosis of sarcoma and entry onto the study, despite equivalent response rates, and (d) increased survival time with liposarcoma or synovial sarcoma, a decreased survival time with malignant fibrous histiocytoma, a lower response rate with leiomyosarcoma, and a higher response rate with liposarcoma (P < .05 for all log-rank and chi2 tests). The Cox model selected good performance status (P < .0001), absence of liver metastases (P = .0001), low histopathologic grade (P = .0002), long time lapse since initial diagnosis (P = .0004), and young age (P = .0045) as favorable prognostic factors of survival time. The logistic model selected absence of liver metastases (P < .0001), young age (P = .0024), high histopathologic grade (P = .0051), and liposarcoma (P = .0065) as favorable prognostic factors of response rate. CONCLUSION: This analysis demonstrates that for advanced soft tissue sarcoma, response to chemotherapy is not predicted by the same factors as is overall survival time. This needs to be taken into account in the interpretation of trials assessing the value of new agents for this disease on the basis of response to treatment. PMID- 10458229 TI - Desmoid tumor: prognostic factors and outcome after surgery, radiation therapy, or combined surgery and radiation therapy. AB - PURPOSE: To evaluate the therapeutic value of resection and the potential benefits of and indications for adjuvant and definitive radiation therapy for desmoid tumors. MATERIALS AND METHODS: We performed a retrospective review of 189 consecutive cases of desmoid tumor treated with surgical resection, resection and radiation therapy, or radiation therapy alone. Treatment was surgery alone in 122 cases, surgery and radiation therapy in 46, and radiation therapy alone in 21. Median follow-up was 9.4 years. RESULTS: Overall, 5- and 10-year actuarial relapse rates were 30% and 33%, respectively. Uncorrected survival rates were 96%, 92%, and 87% at 5, 10, and 15 years, respectively. For the patients treated with surgery, the actuarial relapse rates were 34% and 38% at 5 and 10 years, respectively. Among 78 patients with negative margins, the 10-year recurrence rate was 27%, whereas 40 margin-positive patients had a 10-year relapse rate of 54% (P = .003). Tumors located in an extremity also had a poorer prognosis than did those in the trunk. For patients treated with radiation therapy for gross disease, the 10-year actuarial relapse rate was 24%. For patients treated with combined resection and radiation therapy, the 10-year actuarial relapse rate was 25%. The addition of radiation therapy offset the adverse impact of positive margins seen in the surgical group. CONCLUSION: Wide local excision with negative pathologic margins is the treatment of choice for most desmoid tumors. Function sparing resection is appropriate because adjuvant radiation therapy can offset the adverse impact of positive margins. Unresectable disease should be treated with definitive radiation therapy. PMID- 10458230 TI - Pretreatment nomogram for prostate-specific antigen recurrence after radical prostatectomy or external-beam radiation therapy for clinically localized prostate cancer. AB - PURPOSE: To present nomograms providing estimates of prostate-specific antigen (PSA) failure-free survival after radical prostatectomy (RP) or external-beam radiation therapy (RT) for men diagnosed during the PSA era with clinically localized disease. PATIENTS AND METHODS: A Cox regression multivariable analysis was used to determine the prognostic significance of the pretreatment PSA level, 1992 American Joint Committee on Cancer (AJCC) clinical stage, and biopsy Gleason score in predicting the time to posttherapy PSA failure in 1,654 men with T1c,2 prostate cancer managed with either RP or RT. RESULTS: Pretherapy PSA, AJCC clinical stage, and biopsy Gleason score were independent predictors (P < .0001) of time to posttherapy PSA failure in patients managed with either RP or RT. Two year PSA failure rates derived from the Cox regression model and bootstrap estimates of the 95% confidence intervals are presented in the format of a nomogram stratified by the pretreatment PSA, AJCC clinical stage, biopsy Gleason score, and local treatment modality. CONCLUSION: Men at high risk (> 50%) for early (< or = 2 years) PSA failure could be identified on the basis of the type of local therapy received and the clinical information obtained as part of the routine work-up for localized prostate cancer. Selection of these men for trials evaluating adjuvant systemic and improved local therapies may be justified. PMID- 10458231 TI - Impaired testicular function in patients with carcinoma-in-situ of the testis. AB - PURPOSE: To elucidate the biologic association between germ cell neoplasia and testicular dysfunction, through investigation of Leydig cell function and semen quality in men with carcinoma-in-situ (CIS) of the testis. PATIENTS AND METHODS: We examined two groups of men, unilaterally orchidectomized for testicular cancer. Biopsy of the contralateral testis had showed CIS in a group of 24 patients and no evidence of CIS in the other group of 30 patients. Semen quality and serum levels of testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were compared in these two groups of men after orchidectomy but before further treatment for testicular cancer. RESULTS: Significantly higher LH levels (median, 8.1 IU/L v 4.8 IU/L; P < .001) and generally lower testosterone levels (median, 12.5 nmol/L v 15.5 nmol/L; P = .13) were found in the CIS group. The proportion of patients with Leydig cell dysfunction was higher in the group of patients with CIS (11 of 24) than in the group of patients without (two of 30) (P = .01). Sperm concentration and total sperm count were significantly lower (P < .001) in patients with CIS (median, 0.03 x 10(6)/mL and 0.10 x 10(6), respectively) than in patients without (median, 9.1 x 10(6)/mL and 32 x 10(6), respectively), whereas the levels of FSH were significantly higher (P < .001) in the former group of men (median, 19.6 IU/L v 9.0 IU/L). CONCLUSION: Not only spermatogenesis but also Leydig cell function is impaired in testes with CIS. This impairment could be due to common factors in the pathogenesis of germ cell neoplasm and testicular dysfunction. Alternatively, CIS cells may have a negative impact on Leydig cell function. PMID- 10458232 TI - Chemotherapy dose-intensification for pediatric patients with Ewing's family of tumors and desmoplastic small round-cell tumors: a feasibility study at St. Jude Children's Research Hospital. AB - PURPOSE: To evaluate the feasibility of dose-intensification for patients with Ewing's family of tumors (EFT) and desmoplastic small round-cell tumors. PATIENTS AND METHODS: From February 1992 to June 1996, we treated 53 consecutive patients on our Ewing's protocol. Induction comprised three cycles of ifosfamide/etoposide on days 1 to 3 and cyclophosphamide (CTX)/doxorubicin on day 5, followed by granulocyte colony-stimulating factor. Local control using surgery and/or radiotherapy started at week 9 along with vincristine/dactinomycin. Maintenance included four alternating cycles of ifosfamide/etoposide and doxorubicin/CTX, with randomization to one of two CTX dose levels to determine the feasibility of dose-intensification during maintenance. RESULTS: Patients had a median age of 13.4 years (range, 4.5 to 24.9 years); 34 patients were male and 43 patients were white. Nineteen patients presented with metastatic disease, 29 had tumors greater than 8 cm in diameter, and 26 had primary bone tumors. These patients received 155 induction cycles, 91% of which resulted in grade 4 neutropenia, 68% in febrile neutropenia, and 68% in grade 3 to 4 thrombocytopenia. During maintenance, grade 4 neutropenia and grade 3 to 4 thrombocytopenia occurred in 81% and 85% of cycles, respectively. Thirty-five patients (66%) completed all therapy, only 13 without significant delays; three developed secondary myeloid malignancies. The toxicity and time to therapy completion were similar in both CTX arms. Estimated 3-year survival and event-free survival were 72%+/-8% and 60%+/-9%, respectively. CONCLUSION: Although intensifying therapy seems feasible for 25% of patients on this study, toxicity was considerable. Therefore, the noninvestigational use of dose-intensification in patients with EFT should await assessment of its impact on disease-free survival. PMID- 10458233 TI - Childhood acute lymphoblastic leukemia with the MLL-ENL fusion and t(11;19)(q23;p13.3) translocation. AB - PURPOSE: To determine the molecular characteristics, clinical features, and treatment outcomes of children with acute lymphoblastic leukemia (ALL) and the t(11;19)(q23,p13.3) translocation. PATIENTS AND METHODS: A retrospective analysis of leukemic cell karyotypes obtained from patients with new diagnoses of ALL who were treated at St. Jude Children's Research Hospital or by the Pediatric Oncology Group was performed to identify cases with the t(11;19)(q23;p13.3) translocation. Molecular analyses were performed on these cases to determine the status of the MLL gene and the presence of the MLL-ENL fusion transcript. RESULTS: Among 3,578 patients with ALL and successful cytogenetic analysis, we identified 35 patients with the t(11;19)(q23;p13.3) translocation: 13 infants and 11 older children had B-precursor leukemia, whereas 11 patients had a T-cell phenotype. Although all of the cases examined had MLL rearrangements and MLL-ENL fusion transcripts, outcome varied according to age and immunophenotype. Among B precursor cases, only two of the 13 infants remain in complete remission, compared with six of the 11 older children. Most strikingly, no relapses have occurred among B-precursor patients 1 to 9 years of age or among T-cell patients. CONCLUSION: Although MLL gene rearrangements are generally associated with a dismal outcome in ALL, two distinct subsets with MLL-ENL fusions have an excellent prognosis. Our results suggest that patients with this genetic abnormality who have T-cell ALL or are 1 to 9 years of age should not be considered candidates for hematopoietic stem-cell transplantation during their first remission. PMID- 10458234 TI - Allogeneic bone marrow transplantation versus chemotherapy for the treatment of childhood acute lymphoblastic leukemia in second remission: a single-institution study. AB - PURPOSE: A retrospective analysis of the treatment of childhood acute lymphoblastic leukemia (ALL) in second remission (CR2) was undertaken at our institution to compare the outcome and prognostic factors of patients treated with chemotherapy or allogeneic bone marrow transplantation (BMT). PATIENTS AND METHODS: Seventy-five children who suffered a medullary relapse and achieved a second remission were treated with either an unmodified allogeneic HLA-matched sibling BMT after hyperfractionated total body irradiation (TBI) and cyclophosphamide (n = 38) or chemotherapy according to institutional chemotherapy protocols (n = 37). To avoid the bias of survival from the attainment of second remission in favor of BMT, the final comparative statistical analysis used the landmark approach and comprised 37 and 29 patients from the BMT and chemotherapy groups, respectively RESULTS: The disease-free survival (DFS) rate was 62% and 26% at 5 years, respectively, for the BMT and the chemotherapy groups (P = .03), with relapse rates of 19% and 67%, respectively, for these two groups (P = .01). There was an overall advantage for the BMT therapeutic approach, as compared with chemotherapy, for patients with ALL in CR2 (1) for patients with a WBC count (at diagnosis) of 20 x 10(9)/L or higher (DFS, 40% v 0%) and those with a WBC count of less than 20 x 10(9)/L (DFS, 73% v35%), (2) for patients whose duration of CR1 was less than 24 months (DFS 48% v 9%) and for patients whose duration of CR1 was 24 months or longer (DFS, 81% v 37%) and (3) for patients who were initially treated with intensive regimens incorporating more than five chemotherapy agents (DFS, 57% v 20%) and for patients treated with five agents or fewer (DFS, 72% v 32%). CONCLUSION: In our single-institution series, unmodified HLA-matched allogeneic sibling transplants using hyperfractionated TBI and cyclophosphamide for patients with ALL in CR2 have resulted in superior outcome with a significantly improved probability of DFS and a lower relapse rate, as compared with those for patients treated with chemotherapy, regardless of the duration of first remission, the disease characteristics at diagnosis, or the intensity of prior treatment during first remission. PMID- 10458235 TI - Molecular and clinical remissions in multiple myeloma: role of autologous and allogeneic transplantation of hematopoietic cells. AB - PURPOSE: To describe molecular monitoring of minimal residual disease in patients with myeloma who have achieved complete remission (CR) after autologous or allogeneic transplantation of hematopoietic cells. MATERIALS AND METHODS: Clonal markers based upon the rearrangement of immunoglobulin heavy-chain genes were generated for each patient and used for polymerase chain reaction (PCR) detection of residual myeloma cells. Fifty-one patients entered the program and 36 achieved CR. After transplantation, molecular monitoring was performed on 29 patients (15 autologous and 14 allogeneic transplants) who had molecular markers. RESULTS: Our data show that molecular remissions are rarely achieved (7%) with high-dose chemotherapy followed by single or double autografting. In addition, virtually all peripheral blood progenitor cell and bone marrow samples contained residual myeloma cells, even when sample collection was scheduled after repeated courses of high-dose chemotherapy. All patients autografted with PCR-positive cells remain positive, and eight of 15 have relapsed. Two patients were autografted with PCR-negative cells: one is in clinical and molecular remission, and one relapsed 25 months after the transplant. In the allografting setting, a higher proportion of patients (50%) achieved molecular remission; there were two relapses, one in the PCR-positive group and one in the PCR-negative group. CONCLUSION: This is the first large study of molecular remissions in myeloma patients to use a PCR-based approach utilizing patient-specific tumor markers. The sizeable fraction of patients who achieved molecular remission after allografting with peripheral blood progenitor cells represents a promising finding in an incurable disease. PMID- 10458236 TI - Patterns of outcome following recurrence after myeloablative therapy with autologous bone marrow transplantation for follicular lymphoma. AB - PURPOSE: To assess the patterns of recurrence, management, and survival following recurrence after myeloablative therapy with autologous bone marrow transplantation (ABMT) in patients with follicular lymphoma (FL). PATIENTS AND METHODS: Between June 1985 and October 1995, 99 patients with FL received cyclophosphamide and total-body irradiation with ABMT as consolidation of second or subsequent remission. RESULTS: Median length of follow-up was 5 1/2 years, and 33 patients developed recurrent lymphoma a median of 14 months after ABMT. In 26 patients, the recurrence was overt; in seven, it was detected on surveillance investigation. Twenty-six patients presented with recurrence at previous sites of disease. Twenty-two patients (67%) had FL at the time of recurrence; in 11 (33%), there was evidence of transformation to diffuse large B-cell lymphoma. Eight patients were managed expectantly; five were alive 21 to 53 months later. Twenty five patients have required treatment to date; eight remained alive 6 months to 10 years later, and five were in remission. The Kaplan-Meier estimate of patients alive 5 years after recurrence is 45% (95% confidence interval, 27% to 62%). In univariate and multivariate analyses, survival after recurrence and overall survival after diagnosis were similar to those of a historical control group who received conventional treatment, before the introduction of myeloablative therapy (adjusted hazard ratio [HR], 1.56, P = .3, and HR, 1.34, P = .4, respectively). CONCLUSION: The survival pattern of patients with FL following recurrence after myeloablative therapy and ABMT suggests that this treatment does not compromise outcome in patients in whom it fails, reflecting the survival pattern of the disease when treated conventionally. PMID- 10458237 TI - Comparison of high-dose therapy and autologous stem-cell transplantation with conventional therapy for Hodgkin's disease induction failure: a case-control study. Societe Francaise de Greffe de Moelle. AB - PURPOSE: To determine the prognostic factors and outcome of first-line induction failure Hodgkin's disease patients who were treated with a salvage regimen of high-dose chemotherapy and autologous stem-cell transplantation, and to compare them with matched, conventionally treated patients. PATIENTS AND METHODS: We retrospectively analyzed data relating to 86 Hodgkin's disease patients who underwent autologous stem-cell transplantation after failure of the first chemotherapy regimen, either because they did not enter a complete remission and experienced progression of disease less than 3 months after the end of their first-line treatment or because they showed evidence of disease progression during first-line therapy. Graft patients were matched with 258 conventionally treated patients (three controls per case) for age, sex, clinical stage, B symptoms, and time at risk; patient data were obtained from international databases. RESULTS: Among the 86 graft patients, the median age at diagnosis was 29 years (range, 14 to 57 years). Thirty-nine percent of patients had stage II disease, 23% had stage III disease, and 38% had stage IV disease. Seventy percent of the patients received chemotherapy and 30% received combined modality therapy; 60% of the patients received a seven- or eight-drug regimen. After this first line treatment, 91% had disease progression and 9% had a brief partial response. Eighty patients received a second-line treatment; pretransplantation status was as follows: 24% of patients had a complete remission, 38% had a partial remission (PR), 14% had stable disease, and disease progression occurred in 24%. With a median follow-up of 22 months (range, 4 to 105 months) from diagnosis, the 5-year event-free survival and overall survival rates from transplantation were 25% and 35% (95% confidence intervals, 15 to 36 and 23 to 49), respectively. In multivariate analysis, the pretransplantation disease status after salvage therapy was the only significant prognostic factor for survival (PR: relative risk = 2.8, P = .017; progressive disease: relative risk (RR) = 5.26, P < .001). From diagnosis, the 6-year overall survival rates of the graft patients and 258 matched conventionally treated patients were 38% and 29%, respectively (P = .058). CONCLUSION: Autologous stem-cell transplantation represents the best therapeutic option currently available for patients with primary induction failure and is associated with acceptable toxicity. Response to second-line treatment before high-dose chemotherapy is the only prognostic factor that can be correlated with survival. PMID- 10458238 TI - Mantle irradiation alone for clinical stage I-II Hodgkin's disease: long-term follow-up and analysis of prognostic factors in 261 patients. AB - PURPOSE: To evaluate mantle radiotherapy (MRT) alone as the initial therapy of patients with clinical stage (CS) I-II Hodgkin's disease (HD). PATIENTS AND METHODS: We performed a retrospective study of patients treated with MRT alone for CS I-II supradiaphragmatic HD between 1969 and 1994. Prognostic factor analysis was performed for progression-free survival (PFS) and overall survival (OS). Outcome was also assessed in favorable cohorts defined in the literature. RESULTS: There were 261 eligible patients. The median follow-up period for surviving patients was 8.4 years (range, 1.8 to 27.4 years). The 10-year OS rate was 73%. Multifactor analysis for OS showed that age was the only important prognostic factor. The 10-year PFS rate was 58%. On multifactor analysis for PFS, the most important prognostic factors were clinical stage, B symptoms, histology, number of sites, and tumor bulk. The 10-year PFS rate for lymphocyte-predominant disease was 81% for stage I and 78% for stage II. In favorable patient cohorts defined in the literature, the 10-year PFS rate ranged from 70% to 73% for the whole group and from 71% to 90% in patients with favorable stage I disease, but only from 48% to 57% in patients with favorable stage II disease. On competing risks analysis, the cumulative 10-year incidence of first site of failure in the para-aortic/splenic region alone was 10.5%. Sixty percent of relapsed patients remain progression-free at 10 years after chemotherapy salvage. CONCLUSION: These results support the use of MRT alone in patients with favorable CS I HD and CS I II HD with lymphocyte-predominant histology. The remainder of patients with CS I II HD require more intensive treatment. PMID- 10458239 TI - Laparotomy versus no laparotomy in the management of early-stage, favorable prognosis Hodgkin's disease: a decision analysis. AB - PURPOSE: To perform a decision analysis that compared the life expectancy and quality-adjusted life expectancy of early-stage, favorable-prognosis Hodgkin's disease (HD) managed with and without staging laparotomy, incorporating data on treatment outcomes of HD in the modern era. METHODS: We constructed a decision analytic model to compare laparotomy versus no laparotomy staging for a hypothetical cohort of 25-year-old patients with clinical stages I and II, favorable-prognosis HD. Markov models were used to simulate the lifetime clinical course of patients, whose prognosis depended on the true pathologic stage and initial treatment. The baseline probability estimates used in the model were derived from results of published studies. Quality-of-life adjustments for procedures and treatments, as well as the various long-term health states, were incorporated. RESULTS: The life expectancy was 36.67 years for the laparotomy strategy and 35.92 years for no laparotomy, yielding a net expected benefit of 0.75 years for laparotomy staging. The corresponding quality-adjusted life expectancies for the two strategies were 35.97 and 35.38 quality-adjusted life years (QALYs), respectively, resulting in a net expected benefit of laparotomy staging of 0.59 QALYs. Sensitivity analysis showed that the decision of laparotomy versus no laparotomy was most heavily influenced by the quality-of life weight assigned to the postlaparotomy state. CONCLUSION: Our model predicted that on average, for a 25-year-old patient, proceeding with staging laparotomy resulted in a gain in life expectancy of 9 months, or 7 quality-adjusted months. These results suggest that a role remains for surgical staging in the management of early-stage HD. PMID- 10458240 TI - Hodgkin's disease survivors more fatigued than the general population. AB - PURPOSE: To estimate the level of fatigue and frequency of fatigue cases among Hodgkin's disease survivors (HDS) and compare them with normative data from the general population. PATIENTS AND METHODS: A cross-sectional follow-up study was done of 557 HDS (age range, 19 to 74 years) treated at the Norwegian Radium Hospital from 1971 to 1991. The sample was approached by mail, and their data were compared with normative data from 2,214 controls (age range, 19 to 74 years) representative of the general Norwegian population. Of the 557 HDS, 459 (82%) responded. The mean age (+/- SD) at the time of study was 44+/-12 years, and the mean observation time was 12+/-6 years. The Fatigue Questionnaire (11 items) measures physical and mental fatigue. Two systems of scoring were used, dichotomized (0, 0, 1, and 1) and Likert (0, 1, 2, and 3). Total fatigue (TF) constitutes the sum of all the Likert scores. Caseness was defined as a total dichotomized score of > or =4 and fatigue that lasted 6 months or longer. RESULTS: The HDS had significantly higher levels of TF than the controls (14.3 v 12.2) (P < .001). Fatigue among the HDS equaled that of the controls in poorest health. More HDS (61%) than controls (31%) reported fatigue symptoms lasting 6 months or longer (P < .001). Fatigue cases were more frequent among HDS (men, 24%; women, 27%) than among the controls (men, 9%; women, 12%) (P < .001). Disease stage/substage IB/IIB predicted fatigue caseness (P = .03). No significant associations were found between treatment characteristics and fatigue. CONCLUSION: Hodgkin's disease survivors are considerably more fatigued than the general population and report fatigue of a substantially longer duration. PMID- 10458241 TI - Prospective randomized trial of melphalan and prednisone versus vincristine, carmustine, melphalan, cyclophosphamide, and prednisone in the treatment of primary systemic amyloidosis. AB - PURPOSE: Primary systemic amyloidosis is an immunoglobulin deposition disorder in which insoluble light chains cause organ dysfunction and death. The established conventional therapy is treatment with melphalan and prednisone. We investigated whether treatment with multiple alkylating agents improved the response rate or survival time, compared with melphalan and prednisone therapy. PATIENTS AND METHODS: We treated 101 patients with biopsy-proven primary amyloidosis. The patients were randomly assigned to receive melphalan and prednisone (52 patients) or vincristine, carmustine, melphalan, cyclophosphamide, and prednisone (49 patients). Patients were stratified according to the presence of cardiac involvement, time from diagnosis to randomization, serum beta2-microglobulin level, and whether peripheral neuropathy was the major manifestation of the disease. RESULTS: The median duration of survival after randomization was 29 months, with no differences in survival time between the two groups. There were 29 patients who fulfilled the response criteria: 15 in the vincristine, carmustine, melphalan, cyclophosphamide, and prednisone arm and 14 in the melphalan and prednisone arm. CONCLUSION: Therapy with multiple alkylating agents did not result in a higher response rate or longer survival time, compared with standard melphalan and prednisone treatment in patients with primary systemic amyloidosis. PMID- 10458242 TI - Treatment of patients with low-grade B-cell lymphoma with the combination of chimeric anti-CD20 monoclonal antibody and CHOP chemotherapy. AB - PURPOSE: To determine the safety and efficacy of the combination of the chimeric anti-CD20 antibody, Rituxan (Rituximab, IDEC-C2B8; IDEC Pharmaceuticals Corporation, San Diego, CA), and cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy. PATIENTS AND METHODS: Forty patients with low grade or follicular B-cell non-Hodgkin's lymphoma received six infusions of Rituxan (375 mg/m2 per dose) in combination with six doses of CHOP chemotherapy. RESULTS: The overall response rate was 95% (38 of 40 patients). Twenty-two patients experienced a complete response (55%), 16 patients had a partial response (40%), and two patients, who received no treatment, were classified as nonresponders. Medians for duration of response and time to progression had not been reached after a median observation time of 29 + months. Twenty-eight of 38 assessable patients (74%) continued in remission during this median follow-up period. The most frequent adverse events attributable to CHOP were alopecia (38 patients), neutropenia (31 patients), and fever (23 patients). The most frequent events attributed to Rituxan were fever and chills, observed primarily with the first infusion. No quantifiable immune response to the chimeric antibody was detected. In a subset of 18 patients, the bcl-2 [t(14;18)] translocation was positive in eight patients; seven of these patients had complete remissions and converted to polymerase chain reaction (PCR) negativity by completion of therapy. CONCLUSION: This is the first report demonstrating the safety and efficacy of Rituxan anti-CD20 chimeric antibody in combination with standard-dose systemic chemotherapy in the treatment of indolent B-cell lymphoma. The clinical responses suggest an additive therapeutic benefit for the combination with no significant added toxicity. The conversion of bcl-2 from positive to negative by PCR in blood and/or marrow suggests possible clearing of minimal residual disease not previously demonstrated by CHOP chemotherapy alone. PMID- 10458243 TI - Prognostic significance of magnetic resonance imaging of femoral marrow in patients with myelodysplastic syndromes. AB - PURPOSE: To investigate whether the abnormalities observed on femoral marrow magnetic resonance images are related to the development of leukemia and survival of patients with myelodysplastic syndromes (MDS). PATIENTS AND METHODS: The findings on magnetic resonance images of the femoral marrow were evaluated over periods of 1 to 92 months (median, 18 months) in 42 consecutive adult patients with newly diagnosed MDS. Magnetic resonance images were obtained by the T1 weighted spin echo method and the short T1 inversion recovery technique. RESULTS: Magnetic resonance images showed that the femoral marrow patterns changed from fatty, faint, or nodular to scattered or uniform as the disease progressed. Development of acute myeloid leukemia was observed in only 13 patients whose marrow exhibited a scattered or uniform pattern. The overall survival of the 29 patients with a scattered or uniform marrow pattern was significantly shorter than that of the 13 patients with a fatty, faint, or nodular marrow pattern (10.7% v 73.3% at 7 years; P < .01). The period of leukemia-free survival was also significantly shorter in the patients with a scattered or uniform marrow pattern versus a fatty, faint, or nodular pattern (37.7% v 100% at 7 years; P < .01). CONCLUSION: Magnetic resonance images of the femoral marrow can provide valuable information for assessing the prognosis and determining the most appropriate management of patients with MDS. PMID- 10458244 TI - Treatment of Philadelphia chromosome-positive early chronic phase chronic myelogenous leukemia with daily doses of interferon alpha and low-dose cytarabine. AB - PURPOSE: To evaluate the efficacy of the combination of interferon alpha (IFN alpha) and daily low-dose cytarabine (ara-C) in the treatment of patients with early chronic-phase chronic myelogenous leukemia (CML) (within 1 year of diagnosis). Improving the degree of hematologic and cytogenetic response in patients with Philadelphia chromosome (Ph)-positive CML may improve prognosis. Both IFN-alpha and ara-C induce cytogenetic responses as single-agent therapy in CML. PATIENTS AND METHODS: One hundred forty patients with Ph-positive early chronic-phase CML received subcutaneous injections of IFN-alpha 5 megaunits/m2 daily and ara-C 10 mg daily. Their median age was 46 years; 53% had good-risk disease, 33% had intermediate-risk disease, and 14% had poor-risk disease. Their results were compared with those of patients receiving IFN-alpha with or without intermittent ara-C (7 days/mo). RESULTS: A complete hematologic response (CHR) was achieved in 92% of patients. A cytogenetic response was seen in 74%: it was major in 50% (Ph-positive < 35%) and complete in 31% (Ph-positive 0%). With a median follow-up of 42 months, the 4-year estimated survival rote was 70% (95% confidence interval, 61% to 79%). Significant side effects included fatigue (43%; grade 3/4, 11%), weight loss (19%; grade 3/4, 11%), muscle and bone aches (20%; grade 3/4, 7%), oral ulcers (4%), diarrhea (6%), and neurologic changes (27%, grade 3/4, 6%). The median dose of IFN-alpha was 3.7 megaunits/m2 daily, mainly because of reductions for myelosuppression (70% of cases); the median ara-C dose was 7.5 mg daily. Prognostic risk groups were predictive for response to the IFN alpha plus ara-C combination. The incidence of CHR was higher with IFN-alpha plus daily ara-C compared with IFN-alpha plus intermittent ara-C and IFN-alpha alone (no ara-C) (92% v 84% v 80%, P = .01), as were the incidences of cytogenetic response (74% v 73% v 58%; P = .003) and major cytogenetic response (50% v 38% v 38%; P = .06). The median time to achievement of major cytogenetic response was significantly shorter than that for previous IFN-alpha regimens (7 v 10 v 12 months; P < .01). However, with the present follow-up, the survival and time to blastic transformation were similar. CONCLUSION: The combination of IFN-alpha plus daily low-dose ara-C seems to be promising for the treatment of CML. High rates of CHR and cytogenetic response were observed with acceptable toxicity and a lower daily dose of IFN-alpha compared with our previous studies. PMID- 10458246 TI - Bone marrow transplant patients with life-threatening organ failure: when should treatment stop? AB - PURPOSE: To discuss issues surrounding life support in bone marrow transplant (BMT) patients, issues that may determine how far we go to keep a deteriorating BMT patient alive--and when we stop trying. How can we define survival chance in BMT patients, and when should prolongation of life be deemed inappropriate? Who should make the decision to terminate support? And how should life support be terminated? DESIGN: Prognostic factors that predict for almost certain nonsurvival have been identified in BMT patients with life-threatening organ failure. The concept of futility raises the question of how low the chance of survival must be before termination of life support is justified--but the concept is flawed, and the value judgments involved in decision making must also be considered. Then, once a decision is made, the manner of withholding or withdrawing life support is also open to discussion. CONCLUSION: Despite controversies, there are areas in which improvements to current practice might be considered. More data are required to determine survival chances of BMT patients with life-threatening organ failure. Greater attention might be devoted, in pretransplant counseling, to issues of intensive life support, with the patient's own views being ascertained before transplantation. And, because technologic possibilities are now imposing fewer boundaries, the problem of finite resources may need to be readdressed, with treatment limits being set down before transplantation. PMID- 10458245 TI - CD56 expression in acute promyelocytic leukemia: a possible indicator of poor treatment outcome? AB - PURPOSE: Blast expression of CD56 is frequent in patients with t(8;21)(q22;q22) acute myeloid leukemia and is associated with an inferior outcome. The expression of CD56 has rarely been reported in acute promyelocytic leukemia (APL) and has not been clinically characterized. Therefore, we examined the prognostic significance of CD56 expression in APL. PATIENTS AND METHODS: We identified all reported cases of CD56+ APL in the medical literature and collected clinical, biologic, and therapeutic details. RESULTS: Data were obtained for 12 patients with CD56+ APL (> 20% blast expression of CD56), including four cases from a single institution with a total of 42 APL patients. All of the CD56+ APL patients had documented cytogenetic presence of t(15;17), and of the nine reported isotypes, eight (89%) were S-isoform. Only six CD56+ patients (50%) attained complete remission (CR); the other six individuals died within 35 days of presentation. Of the six patients who attained a CR, three (50%) relapsed at 111, 121, and 155 weeks, whereas three remained in continuous CR at 19, 90, and 109 weeks. Comparison of the control CD56- to CD56+ APL patients demonstrated that the latter group had a significantly lower fibrinogen level (P = .007), and among patients for whom data were available, there was a higher frequency of the S isoform (P = .006). Additionally, the CR rate (50% v 84%, P = .025) and overall median survival (5 v 232 weeks; P = .019) were significantly inferior for CD56+ APL patients. CONCLUSION: CD56+ acute promyelocytic leukemia is infrequent, seems to occur more frequently with the S-isoform subtype, and may be associated with a lower CR rate and inferior overall survival. PMID- 10458247 TI - Polymerase chain reaction-based detection of circulating melanoma cells as an effective marker of tumor progression. Melanoma Cooperative Group. AB - PURPOSE: Reverse transcriptase (RT) polymerase chain reaction (PCR) with multiple markers has been demonstrated to be highly sensitive in detecting circulating cells from patients with malignant melanoma (MM). We evaluated the clinical significance of the presence in peripheral blood of specific PCR-positive mRNA markers as an expression of circulating melanoma cells. PATIENTS AND METHODS: Total cellular RNA was obtained from the peripheral blood of 235 patients with either localized (n = 154) or metastatic (n = 81) melanoma. We performed RT-PCR using tyrosinase, p97, MUC18, and MelanA/MART1 as gene markers. The PCR products were analyzed by gel electrophoresis and Southern blot hybridization. In addition, 20 healthy subjects and 21 patients with nonmelanoma cancer were used as negative controls. RESULTS: Although detected at various levels among assessable patients, each mRNA marker was significantly correlated with disease stage. A significant correlation with disease stage was demonstrated for patients who were positive to all four markers (P < .0001) or to at least three markers (P < .001). Univariate analysis showed a significant correlation between risk of recurrence (evaluated in stage I, II, and III patients) and increasing number of PCR-positive markers (P = .0002). Logistic regression multivariate analysis indicated that each single marker (except tyrosinase) and, more especially, the presence of four PCR-positive markers remained statistically independent prognostic factors for tumor progression. CONCLUSION: Our data establish the existence of a significant correlation among clinical stages, tumor progression, and presence of circulating melanoma-associated antigens in peripheral blood of MM patients. Preliminary assessment of a subset of patients with a higher risk of recurrence needs longer follow-up and further studies to define the role of RT PCR in monitoring MM patients. PMID- 10458248 TI - Decision analysis as an aid to determining the management of early low rectal cancer for the individual patient. AB - PURPOSE: Because there are no data available from randomized controlled trials (RCT), a decision analysis was performed to aid in the decision of which option, a local excision with or without radiotherapy or an abdominal perineal resection (APR), should be offered to medically fit patients with early (suspected T1/T2) low (< 5 cm) rectal cancer. METHODS: All clinically relevant outcomes, including complications of surgery and radiotherapy, cure, salvageability after local recurrence, distant disease, and death, were modeled for both options. The probabilities of complications and outcomes after radiotherapy and/or local excision were derived from weighted averages of results from studies conducted between 1969 and 1997. The probabilities for the APR option were extracted from relevant RCTs. Long- and short-term patient-centered utilities for each complication and outcome were extracted from the literature and from expert opinion. RESULTS: The expected utility of local excision (EU = 0.81) for the base case was higher than the expected utility for APR (EU = 0.78). Although the result was sensitive to all variables, local excision was always favored over APR within the plausible ranges of the variables taken one, two, or three at a time. The model illustrated the tension between the patient's perception of a colostomy and the higher recurrence rates with local excision. CONCLUSION: The results of this decision analysis suggest that local therapy for early low rectal cancer is the preferred method of treatment. However, there must be careful preoperative assessment, patient selection, and consideration of patient concerns. In addition, decision analysis may be useful in providing patient information and assisting in decision making. PMID- 10458249 TI - Phase II study of a combination of irinotecan and cisplatin against metastatic gastric cancer. AB - PURPOSE: A phase II study of a combination chemotherapy regimen of cisplatin (CDDP) and irinotecan (CPT-11) was conducted to assess its efficacy and feasibility in patients with metastatic gastric cancer. PATIENTS AND METHODS: Eligibility criteria included the following: (1) histologically proven gastric cancer with measurable metastatic lesions, (2) performance status of 2 or less, (3) age of 75 years or younger, (4) one or no prior chemotherapy regimens, (5) adequate bone marrow, liver, renal, and cardiac functions, and (6) written informed consent. The treatment consisted of CPT-11 (70 mg/m2) on day 1 and day 15 and CDDP (80 mg/m2) on day 1, repeated every 4 weeks. RESULTS: Forty-four patients were entered onto the study. The overall response rate was 48% (21 of 44 patients, 95% confidence interval [CI], 33% to 63%) and included one complete remission (2%). The response rate of the patients who had not received prior chemotherapy was 59% (17 of 29 patients, 95% CI, 39% to 77%). The median survival time was 272 days for all patients and 322 days for the 29 patients who had not received prior chemotherapy. Grade 4 neutropenia was observed in 25 patients (57%), and grade 3 or 4 diarrhea was observed in nine patients (20%). Other adverse reactions were mild. No treatment-related deaths occurred. CONCLUSION: This combination chemotherapy regimen is active and well tolerated. It may be an appropriate regimen for future phase III trials. PMID- 10458250 TI - Recurrence patterns of hepatocellular and fibrolamellar carcinoma after liver transplantation. AB - PURPOSE: Tumor recurrence is the major limitation of long-term survival after liver transplantation for hepatocellular carcinoma (HCC) or fibrolamellar carcinoma (FLC). Understanding tumor-biologic characteristics is important for selection of patients and for development of adjuvant therapeutic strategies. PATIENTS AND METHODS: The study included 69 patients who underwent potentially curative liver transplantation for HCC/FLC and survived for more than 150 days; minimum follow-up was 33 months. Frequency, localization, and timing of recurrence were analyzed and compared with primary tumor and patient characteristics. RESULTS: Tumor recurrence was observed in 39 patients at 67 locations. Hematogenous spread was the major route of tumor recurrence (87%), and the most frequent sites were the liver (62%), lung (56%), and bone (18%). Parameters associated with recurrence were absence of cirrhosis, tumor size greater than 5 cm, more than five nodules, vascular infiltration, and International Union Against Cancer (UICC) stage IVA. Selective intrahepatic recurrence was found in nine patients (23%); it was associated with highly differentiated tumors, lack of vascular infiltration, and male sex. Recurrence at multiple sites was found predominantly in young patients (< or = 40 years) and for multicentric (> 5) primary tumors. Recurrences were observed within a wide time range after transplantation (43 to 3,204 days; median, 441 days); late recurrences (> 1,000 days, n = 8) were associated with highly differentiated or fibrolamellar tumors and low UICC stages. Surgical treatment was the only therapeutic option associated with prolonged survival after recurrence. CONCLUSION: In transplant recipients, hepatocellular carcinomas vary considerably in their pattern and kinetics of metastases. Tumor cells may persist in a dormant state for long time periods before giving rise to clinical metastases. Surgical treatment of recurrence should be considered whenever possible. PMID- 10458251 TI - Phase I study in cancer patients of a replication-defective avipox recombinant vaccine that expresses human carcinoembryonic antigen. AB - PURPOSE: A phase I clinical trial in patients with advanced carcinoma was conducted, using a replication-defective avipox vaccine containing the gene for the human carcinoembryonic antigen (CEA). The canarypox vector, designated ALVAC, has the ability to infect human cells but cannot replicate. PATIENTS AND METHODS: The recombinant vaccine, designated ALVAC-CEA, was administered intramuscularly three times at 28-day intervals. Each cohort of six patients received three doses of either 2.5 x 10(5), 2.5 x 10(6), or 2.5 x 10(7) plaque-forming units of vaccine. RESULTS: The vaccine was well tolerated at all dose levels and no significant toxicity was attributed to the treatment. No objective antitumor response was observed during the trial in patients with measurable disease. Studies were conducted to assess whether ALVAC-CEA had the ability to induce cytolytic T-lymphocyte (CTL) responses in patients with advanced cancer. Peripheral blood mononuclear cells (PBMCs) from patients with the MHC class I A2 allele were obtained before vaccine administration and 1 month after the third vaccination. Peripheral blood mononuclear cells were incubated with the CEA immunodominant CTL epitope carcinoembryonic antigen peptide-1 and interleukin 2 and quantitated using CTL precursor frequency analysis. In seven of nine patients evaluated, statistically significant increases in CTL precursors specific for CEA were observed in PBMCs after vaccination, compared with before vaccination. CONCLUSION: These studies constitute the first phase I trial of an avipox recombinant in cancer patients. The recombinant vaccine ALVAC-CEA seems to be safe and has been demonstrated to elicit CEA-specific CTL responses. These studies thus form the basis for the further clinical exploration of the ALVAC-CEA recombinant vaccine in phase I/II studies in protocols designed to enhance the generation of human T-cell responses to CEA. PMID- 10458252 TI - Randomized phase II study of the neurokinin 1 receptor antagonist CJ-11,974 in the control of cisplatin-induced emesis. AB - PURPOSE: To determine the efficacy and safety of the neurokinin type 1 receptor antagonist CJ-11,974 for the control of high-dose cisplatin-induced emesis. PATIENTS AND METHODS: A double-blind, randomized, phase II design with a group sequential stopping rule was used in this study. Sixty-one patients with cancer who were receiving cisplatin at a dose of at least 100 mg/m2 for the first time were enrolled. All patients received granisetron 10 microg/kg and dexamethasone 20 mg intravenously 30 minutes before they were given cisplatin. Patients were randomly assigned to two groups: group 1 received CJ-11,974 100 mg, and group 2 received placebo orally 30 minutes before and 12 hours after cisplatin and then twice daily on days 2 through 5 after cisplatin. The primary end point was the percentage of patients who developed delayed emesis (emesis on the second to fifth days after cisplatin). RESULTS: Thirty patients were enrolled in group 1, and 31 patients were enrolled in group 2. Fifty-eight patients were assessable for efficacy. Complete control of emesis (expressed as the percentage of patients who had no emesis) was as follows: day 1, 85.7% (group 1) and 66.7% (group 2) (P = .090); days 2 through 5, 67.8% (group 1) and 36.6% (group 2) (P = .0425, adjusted); days 1 through 5, 64.3% (group 1) and 30% (group 2) (P = .009). Patients in group 1 experienced significantly less nausea than patients in group 2 on day 1 (P = .024). Treatment was well tolerated in both groups. CONCLUSION: We conclude from this exploratory phase II trial that CJ-11,974 is superior to placebo in controlling cisplatin-induced delayed emesis and may provide additive benefit in acute emesis and nausea control when combined with a 5 hydroxytryptamine-3 receptor antagonist and dexamethasone. Additional larger trials are indicated to confirm the clinical value of CJ-11,974. PMID- 10458253 TI - Costs of treating and preventing nausea and vomiting in patients receiving chemotherapy. AB - PURPOSE: To evaluate the effect of ondansetron availability on the costs of managing nausea and vomiting. METHODS: We retrospectively assessed antiemetic costs (drug costs, nursing time, pharmacy time, physician's time, supplies, and facility "hotel" costs, in 1991 Canadian dollars) for all patients who received moderately or highly emetogenic chemotherapy from 6 months before to 6 months after ondansetron became commercially available in September 1991. We compared the costs for treating patients who received ondansetron versus those who received other antiemetic regimens, the costs for treating patients in the 6 months before versus the 6 months after ondansetron commercial availability, and the costs for treating patients in the first 4 months versus the last 4 months of the study period. RESULTS: We found no cost differences for patients treated with ondansetron versus other antiemetic regimens. However, there was a significant reduction in emesis management costs for patients treated after versus before the availability of ondansetron: for patients treated in the last third versus first third of the study period, there was a decrease in cost per patient per month of treatment of $374 (95% confidence interval, $243 to $505). These savings were achieved through a reduction in hospital bed days and other costs associated with the prevention and more effective management of nausea and vomiting. At the same time, the number of patients who received emetogenic chemotherapy and their average age increased, presumably because of the better control of gastrointestinal toxicity. CONCLUSION: Ondansetron availability has been associated with changes in the clinical management of cancer patients receiving chemotherapy and with overall cost savings compared with previously available antiemetic therapy. PMID- 10458254 TI - First year after head and neck cancer: quality of life. AB - PURPOSE: Treatment regimens for head and neck cancer patients profoundly affect several quality-of-life domains. Rehabilitative needs have been identified through cross-sectional analyses; however, few studies have prospectively assessed quality of life, included assessment of psychosocial variables, and identified predictors of long-term follow-up. PARTICIPANTS AND METHODS: The present study addresses these limitations through a prospective assessment of 105 patients with a newly diagnosed first primary squamous cell carcinoma of the oral cavity, pharynx, or larynx. Participants were enrolled onto a larger randomized controlled trial comparing a provider-delivered smoking cessation intervention with a usual-care-advice control condition. Participants completed a battery of self-report measures after diagnosis and before treatment and additional quality of-life instruments at 1 and 12 months after initial smoking cessation advice. RESULTS: Participants displayed improvements at 12 months in functional status (P = .006) and in the areas of eating, diet, and speech; however, the latter three represent areas of continued dysfunction, and the changes were not statistically significant. Despite these improvements, patients reported a decline in certain quality-of-life domains, including marital (P = .002) and sexual functioning (P = .017), as well as an increase in alcohol use (P < .001). Predictors of quality of life at 12 months included treatment type, the Vigor subscale of the Profile of Mood States instrument, and quality-of-life scores obtained 1 month after initial smoking cessation advice. CONCLUSION: Results reinforce the need for rehabilitation management through the integration of psychologic and behavioral interventions in medical follow-up. PMID- 10458255 TI - Implementing guidelines for cancer pain management: results of a randomized controlled clinical trial. AB - PURPOSE: Pain and symptom management is an integral part of the clinical practice of oncology. A number of guidelines have been developed to assist the clinician in optimizing comfort care. We implemented clinical guidelines for cancer pain management in the community setting and evaluated whether these guidelines improved care. PATIENTS AND METHODS: Eighty-one cancer patients, aged 37 to 76 years, were enrolled onto a prospective, longitudinal, randomized controlled study from the outpatient clinic settings of 26 western Washington-area medical oncologists. A multilevel treatment algorithm based on the Agency for Health Care Policy and Research Guidelines for Cancer Pain Management was compared with standard-practice (control) pain and symptom management therapies used by community oncologists. The primary outcome of interest was pain (Brief Pain Inventory); secondary outcomes of interest were all other symptoms (Memorial Symptom Assessment Scale) and quality of life (Functional Assessment of Cancer Therapy Scale). RESULTS: Patients randomized to the pain algorithm group achieved a statistically significant reduction in usual pain intensity, measured as slope scores, when compared with standard community practice (P < .02). Concurrent chemotherapy and patient adherence to treatment were significant mediators of worst pain. There were no significant differences in other symptoms or quality of life between the two treatment groups. CONCLUSION: This guideline implementation study supports the use of algorithmic decision making in the management of cancer pain. These findings suggest that comprehensive pain assessment and evidence based analgesic decision-making processes do enhance usual pain outcomes. PMID- 10458256 TI - Can 40 seconds of compassion reduce patient anxiety? AB - PURPOSE: To use a standardized videotape stimulus to assess the effect of physician compassion on viewers' anxiety, information recall, treatment decisions, and assessment of physician characteristics. PARTICIPANTS AND METHODS: One hundred twenty-three healthy female breast cancer survivors and 87 women without cancer were recruited for this study. A randomized pretest/posttest control group design with a standardized videotape intervention was used. Participants completed the State-Trait Anxiety Inventory (STAI), an information recall test, a compassion rating, and physician attribute rating scales. RESULTS: Women who saw an "enhanced compassion" videotape rated the physician as warmer and more caring, sensitive, and compassionate than did women who watched the "standard" videotape. Women who saw the enhanced compassion videotape were significantly less anxious after watching it than the women in the other group. Nevertheless, information recall was relatively low for both groups, and enhanced compassion did not influence patient decisions. Those who saw the enhanced compassion videotape rated the doctor significantly higher on other positive attributes, such as wanting what was best for the patient and encouraging the patient's questions and involvement in decisions. CONCLUSION: The enhanced compassion segment was short, simple, and effective in decreasing viewers' anxiety. Further research is needed to translate these findings to the clinical setting, where reducing patient anxiety is a therapeutic goal. PMID- 10458257 TI - RET proto-oncogene in the development of human cancer. AB - The RET proto-oncogene, located on chromosome subband 10q11.2, encodes a receptor tyrosine kinase expressed in tissues and tumors derived from neural crest. Germline (present in every cell of the body) mutations in RET cause multiple endocrine neoplasia type 2 (MEN 2), an inherited cancer syndrome characterized by medullary thyroid carcinoma (MTC), pheochromocytoma (PC), and hyperparathyroidism (HPT). This knowledge has allowed molecular diagnosis and presymptomatic DNA based testing to become possible. RET testing is considered the standard of care in MEN 2 families because clinical decisions are made based on the results of such gene testing. There appears to be a correlation between specific RET mutation type and organ-specific tumor development. Such knowledge might be useful in tailoring targeted surveillance in the near future. Somatic (in the tumor only) RET mutations have been found in a proportion of sporadic MTCs and PCs. Whether the presence of somatic RET mutation is associated with a poor prognosis is currently being investigated as another tool for molecular medicine. PMID- 10458258 TI - Chemotherapy and bone marrow transplantation for cancer patients who are also chronic hepatitis B carriers: a review of the problem. AB - In places where hepatitis B virus (HBV) infection is endemic, it is often necessary to give chemotherapy to or perform bone marrow transplantation for cancer patients who are also chronic HBV carriers. When standard chemotherapy was given to lymphoma patients, elevation of liver transaminases was observed in nearly half of those who were chronic HBV carriers. Ten percent of them became jaundiced, and the overall liver-related mortality was about 5%. There is currently no reliable way to predict the severity of HBV reactivation after chemotherapy. The risk is probably higher when the chemotherapy used is significantly immunosuppressive and the viral load in the liver is high. Different strategies have been used in an attempt to reduce the risk of HBV reactivation after chemotherapy, but they have not been very successful. Further studies will be required to determine the impact of newly available antiviral agents that are active against HBV. Recipients who are carriers of HBV or who receive hepatitis B surface antigen (HBsAg)-positive marrow are at increased risk of hepatitis B-related morbidity and mortality after bone marrow transplantation (BMT). There is evidence to suggest that prophylactic use of an active antiviral agent, such as famciclovir, may result in a significant decrease in the incidence and severity of HBV reactivation after BMT. Sustained serologic clearance of chronic HBV infection has also been reported in many HBsAg-positive marrow recipients receiving hepatitis B surface antibody-positive marrow from their allogeneic donors. There seems to be a transfer of both humoral and cellular immunity against HBV from donors to recipients. Further prospective studies are required to define the best approach to manage HBsAg-positive cancer patients receiving chemotherapy or BMT. It is recommended that all cancer patients be checked for their hepatitis B status before receiving chemotherapy or a bone marrow transplant, especially if they reside in or come from endemic areas of HBV infection. PMID- 10458259 TI - B-cell chronic lymphocytic leukemia: a bird of a different feather. AB - PURPOSE: To review the recent major advances in the molecular and cell biology of B-cell chronic lymphocytic leukemia (B-CLL). METHODS: We analyzed the nature of malignant B-CLL B cells and their interactions with the microenvironment. RESULTS: B-CLL is a malignancy of a mantle zone-based subpopulation of anergic, self-reactive, activated CD5+ B cells devoted to the production of polyreactive natural autoantibodies. It is the quintessential example of a human malignancy that primarily involves defects in the induction of programmed cell death. An abnormal karyotype is observed in about 50% of patients with B-CLL. Patients with 13q14 abnormalities show heavy somatic mutation and have a benign disease. Trisomy 12 is associated with unmutated VH genes, atypical cellular morphology, and progressive disease. Extended cell survival is further shielded by a kinetic refractoriness likely promoted by abnormalities of the B-cell antigen receptor complex and favored by some cytokines that highlight a reciprocal dialog between malignant B and T cells. Because the tumor cells act as the major accessory cells, the accumulating malignant B-cell population per se is a hurdle to the production of normal antibodies and leads to a progressive and severe hypogammaglobulinemia. Conceivably, in the presence of certain immunoglobulin genes and when the T-cell control becomes deficient, activated malignant B cells may become able to present self-antigens and drive residual normal B cells to produce polyclonal autoantibodies restricted to self-antigens expressed only by blood cells and cause autoimmune cytopenias. CONCLUSION: The distinctiveness of B CLL B cells explains why B-CLL is different from other B-cell tumors and accounts for the development of immune deficiency and autoimmunity. PMID- 10458260 TI - Review of the comparative pharmacology and clinical activity of cisplatin and carboplatin. AB - PURPOSE: To review the pharmacodynamics, pharmacokinetics, toxicities, and relative clinical activities of cisplatin and carboplatin. Through a search of the MEDLINE database, we identified phase III clinical trials and pharmacologic studies comparing cisplatin and carboplatin published in the English language medical literature from January 1966 to December 1997. RESULTS: Prospective randomized trials comparing cisplatin to carboplatin were identified for ovarian (n = 12), germ cell (n = 4), non-small-cell lung (n = 1), small-cell lung (n = 3), and head and neck (n = 4) cancers. Carboplatin and cisplatin were equally effective in suboptimally debulked ovarian cancer and extensive-stage small-cell lung cancer. One study each showed a trend toward better survival in favor of cisplatin for patients with optimally debulked ovarian and limited-stage small cell lung cancers. These results were, however, based on subset analyses. In germ cell tumors, carboplatin was inferior because of lower relapse-free survival rates. Cisplatin produced superior response rates and survival in head and neck cancers. There are no published randomized phase III studies of bladder, cervical, endometrial, and esophageal cancers. CONCLUSION: Carboplatin does not possess equivalent activity to cisplatin in all platinum-sensitive tumors. Carboplatin can replace cisplatin in chemotherapy regimens for suboptimally debulked ovarian cancer. Two ongoing studies will address the same question in optimally debulked disease. Carboplatin can also be substituted for cisplatin in the treatment of non-small-cell and extensive-stage small-cell lung cancers. Its role in limited-stage small-cell lung cancer needs to be investigated further. Carboplatin is inferior to cisplatin in germ cell, head and neck, and esophageal cancers. Randomized studies are needed to determine whether carboplatin has equivalent efficacy to cisplatin in bladder, cervical, and endometrial cancers. PMID- 10458261 TI - International Consensus Conference on High-Dose Therapy with Hematopoietic Stem Cell Transplantation in Aggressive Non-Hodgkin's Lymphomas: report of the jury. PMID- 10458262 TI - Hemialopecia due to regional chemotherapy. PMID- 10458263 TI - Multifocal cerebral demyelination secondary to fluorouracil and levamisole therapy. PMID- 10458264 TI - Increased HER2 with U.S. Food and Drug Administration-approved antibody. PMID- 10458265 TI - Use of gabapentin to treat taxane-induced myalgias. PMID- 10458266 TI - Future potentials for using osteogenic stem cells and biomaterials in orthopedics. AB - Ideal skeletal reconstruction depends on regeneration of normal tissues that result from initiation of progenitor cell activity. However, knowledge of the origins and phenotypic characteristics of these progenitors and the controlling factors that govern bone formation and remodeling to give a functional skeleton adequate for physiological needs is limited. Practical methods are currently being investigated to amplify in in vitro culture the appropriate autologous cells to aid skeletal healing and reconstruction. Recent advances in the fields of biomaterials, biomimetics, and tissue engineering have focused attention on the potentials for clinical application. Current cell therapy procedures include the use of tissue-cultured skin cells for treatment of burns and ulcers, and in orthopedics, the use of cultured cartilage cells for articular defects. As mimicry of natural tissues is the goal, a fuller understanding of the development, structures, and functions of normal tissues is necessary. Practically all tissues are capable of being repaired by tissue engineering principles. Basic requirements include a scaffold conducive to cell attachment and maintenance of cell function, together with a rich source of progenitor cells. In the latter respect, bone is a special case and there is a vast potential for regeneration from cells with stem cell characteristics. The development of osteoblasts, chondroblasts, adipoblasts, myoblasts, and fibroblasts results from colonies derived from such single cells. They may thus, theoretically, be useful for regeneration of all tissues that this variety of cells comprise: bone, cartilage, fat, muscle, tendons, and ligaments. Also relevant to tissue reconstruction is the field of genetic engineering, which as a principal step in gene therapy would be the introduction of a functional specific human DNA into cells of a patient with a genetic disease that affects mainly a particular tissue or organ. Such a situation is pertinent to osteogenesis imperfecta, for example, where in more severely affected individuals any improvements in long bone quality would be beneficial to the patient. In conclusion, the potentials for using osteogenic stem cells and biomaterials in orthopedics for skeletal healing is immense, and work in this area is likely to expand significantly in the future. PMID- 10458267 TI - Vertebroplasty: clinical experience and follow-up results. AB - This study was undertaken to report the clinical experience with percutaneous minimal invasive vertebroplasty using polymethyl-methacrylcate (PMMA) for a consecutive group of patients. Over the period of the last 4 years, 40 patients were treated at 68 vertebral segment levels with the intention to relieve pain related to vertebral body lesions. Reduced vertebral body height and destruction of the posterior vertebral wall were not considered to be exclusion criterias. The vertebroplasty procedure was performed under general anesthesia and in prone position with imaging control using mostly biplane DSA fluoroscopic guidance, and rarely with single-plane mobile DSA combined with computed tomographic guidance. Unilateral, but more frequently bilateral, transpedicular introduction of a 2-3 mm OD needle was followed by an injection of polymethyl-methacrylcate (PMMA). PMMA preparation involved a diluted mixture (20 mL powder for 5 mL liquid) allowing for an extended polymerization time of up to 8 min. The PMMA was mixed with metallic powder to enhance its radio-opacity. Before PMMA injection, a vertebral phlebography was obtained to evaluate the filling pattern and identify sites of potential PMMA leakage. Injection of opacified PMMA was performed under continuous visual control with fluoroscopy to obtain adequate filling and to avoid important PMMA leakage. Clinical follow-up involved an evaluation using a questionnaire for assessment of pain, pain medication, and mobility. One to six levels were treated in one to three treatment sessions for patients with metastatic, osteoporotic, and hemangiomatous lesions of the vertebral bodies who presented with pain. The results observed matched those reported previously with a success rate of approximately 80% and a complication rate below 6% per treated level. Treatment failure and complications observed were related to leakage, insufficient pretreatment evaluation, anesthesia, or patient position during treatment. Image guidance with fluoroscopy was efficient both for precise transpedicular approach and PMMA implantation control. Vertebroplasty is very efficient for treatment of pain. Treatment failure was mostly related to insufficient pretreatment clinical evaluation, and complication due to excessive PMMA volume injection. Control of PMMA volume seems to be the most critical point for avoiding complications. A good fluoroscopy control is therefore mandatory. PMID- 10458268 TI - Temperature elevation caused by bone cement polymerization during vertebroplasty. AB - Percutaneous vertebroplasty (PVP), whereby polymethylmethacrylate cement is injected into the vertebral body (VB), has been used to successfully treat various spinal lesions. The mechanism responsible for the palliative effect of PVP is unknown, but it may be the result of neural damage caused by heat liberated during polymerization of the polymethylmethacrylate. The purpose of the current study was to measure in vitro temperature histories at three key locations (anterior cortex, center, spinal canal) in VBs injected with one of two different bone cements (Simplex P and Orthocomp) to determine the role temperature plays in PVP. Twelve VBs (T11-L2) from three elderly female spines were instrumented with thermocouples and injected with 10 cc of one of the two cements. Temperatures were measured with the VBs in a bath (37 degrees C) for 15 min after injection. A Student's paired t-test was used to determine differences in peak temperature and time above 50 degrees C between the two cement groups. Peak temperatures and temperatures above 50 degrees C were significantly higher and longer, respectively, at the center of VBs injected with Simplex P (61.8 +/- 12.7 degrees C; 3.6 +/- 2.1 min) than those injected with Orthocomp (51.2 +/- 6.2 degrees C; 1.3 +/- 1.4 min). There was no significant difference in peak temperature between cements at the spinal canal location; temperature there did not rise above 41 degrees C. Although thermal damage to intraosseous neural tissue caused by cement polymerization cannot be ruled out as a potential mechanism for pain relief experienced by patients subsequent to PVP, it seems unlikely based on the worst-case conditions tested in the current study. PMID- 10458270 TI - The effect of monomer-to-powder ratio on the material properties of cranioplastic. AB - Percutaneous vertebroplasty consists of injecting polymethylmethacrylate cement into the cancellous bone of vertebral bodies for the treatment of various lesions of the spine, including osteoporotic compression fractures. Clinicians practicing vertebroplasty commonly alter the mixture of monomer-to-powder recommended by the manufacturer in an effort to decrease viscosity and increase the working time. The purpose of the current study was to measure the effect that varying the monomer-to-powder ratio has on the compressive material properties of a cement (Cranioplastic) commonly used in vertebroplasty. Cylindrical specimens were prepared varying a monomer-to-polymer ratio of 0.40 to 1.07 ml/g and tested per the American Society for Testing and Materials standard F451. Specimens prepared at 0.53 mL/g, which is near the manufacturer's recommended monomer-to-polymer mixture of 0.57 mL/g, exhibited the greatest mean values for ultimate compressive stress, yield stress, and elastic modulus. Specimens prepared at higher or lower ratios exhibited diminished strength, in some cases by as much as 24%. Although altering the monomer-to-powder ratio affects the cement's material properties, it is as yet unknown if the decrease is clinically significant. PMID- 10458269 TI - An in vitro biomechanical evaluation of bone cements used in percutaneous vertebroplasty. AB - The purpose of this study was to determine the strength and stiffness of osteoporotic vertebral bodies (VBs) subjected to compression fractures and subsequently treated with bipedicular injections of various polymethylmethacrylate cements. Ten spines were harvested from nonembalmed female cadavers (age 68.6 +/- 13.7 years) and evaluated for bone mineral density using the dual energy X-ray absorptiometry method (t-score = -2.3 +/- 2.4). The 50 VBs (L1-L5) were disarticulated, compressed in a materials testing machine to determine initial strength and stiffness, and then assigned to one of six groups. Two of these groups (n = 8, n = 9) concerned experimental cements, the results of which are not reported here. The 33 vertebral bodies in the remaining four groups were left untreated or were repaired using a transpedicular injection of one of three commercially available polymethylmethacrylate cements. These four groups were: a) no treatment (no cement, n = 8); b) Simplex P (n = 9); c) Cranioplastic (n = 8); and d) Osteobond (n = 8). The VBs were then compressed again according to the initial protocol, and posttreatment strength and stiffness were measured. Results suggested that bipedicular injection of Simplex P and Osteobond restored VB stiffness to initial values, whereas VBs injected with Cranioplastic were significantly less stiff than in their initial state. VBs injected with cement (regardless of type) were significantly stronger than they were initially. PMID- 10458271 TI - Effects of various adjuvants (lactic acid, glycerol, and chitosan) on the injectability of a calcium phosphate cement. AB - Calcium phosphate cements are well-known orthopedic materials for filling bone. Various formulations are proposed. The current challenge is to place the material in the surgical site by methods as least invasive as possible. One approach consists of making the cement injectable by incorporation of various adjuvants. However, the requirement properties of the cement must be preserved: setting times suited to a convenient delay with surgical intervention, limited disintegration in aqueous medium, and sufficient mechanical resistance. Various additives were studied: in particular, lactic acid, glycerol, chitosan, and sodium glycerophosphate. Injectability, setting time, disintegration, and toughness after 10 days were followed in vitro. Glycerol greatly improved injectability and increased setting time, but decreased mechanical properties. Lactic acid reduced setting time, increased toughness of the material, but limited the dissolution rate. After injection, the cement did not present any disintegration. The effects lactic acid were correlated with the formation of calcium complex. Its association with sodium glycerophosphate is particularly interesting. Chitosan alone improved injectability, increased setting time, and limited the evolution of the cement by maintaining the OCP phase. Only slight disintegration was observed. These first results show that is possible to transform the cement into an injectable paste by addition of adjuvants without fundamentally modifying the chemical reactions occurring during setting and hardening. PMID- 10458272 TI - Volume effect on biological properties of a calcium phosphate hydraulic cement: experimental study in sheep. AB - Injectable calcium phosphate hydraulic cements (CPHC) are a new family of bone substitutes within the class of bone reconstruction biomaterials. In this work, CPHC were tested in two consistencies (preset blocks or liquid paste) in an experimental model of cancellous bone defect in sheep. The defects were eight times larger than those investigated previously in rabbits. Three delays (12, 24, and 52 weeks) were used. Before death, a double label of oxytetracycline and alizarine was made intravenously. The distribution of implants was randomized, histomorphometric evaluation was performed and compared with micrographic observation, and optical microscopy of stained sections was performed either under visible, ultraviolet, or polarized light. The results were compared with spontaneous healing of empty defects and with a control group of normal cancellous bone from sheeps of the same age. No significant difference has been observed between premolded and injected implants. In the sheep model, the degradation and new bone formation rates are three times slower, compared with those observed previously in rabbits. New bone formation increased from 5.9% (12 weeks) up to 11.0% (24 weeks) in the empty defect group. In the cement groups, 28.3% new bone was obtained at 12 weeks, which seemed then to level off (27.8% new bone at 24 weeks). Cement residues appear as radio-opaque cylinders on microradiographs. In all cases, a radiolucent layer was observed at the cement/bone interface at 24 weeks. Stained sections showed the formation of a fibroconnective capsule around the residual cement, which presumably slows down new bone formation. Nevertheless, quantitative bone remodeling was accelerated in the cement group; mineral apposition as well as adjusted apposition rates were higher, and the formation period as well as the mineralization of osteoid tissue were faster compared with empty cavities and controls. These results point to higher osteoblast activity and better exchange with surrounding tissues in the defects filled with cement. PMID- 10458273 TI - Histological and biomechanical studies of two bone colonizable cements in rabbits. AB - We have developed two colonizable bone cements: the first is a partially resorbable bisphenol-alpha-glycidyl methacrylate (Bis-GMA)-based cement (PRC) and the second is a calcium phosphate cement (CPC). PRC is composed of aluminous silanized ceramic and particles of a bioresorbable polymer embedded in a matrix of Bis-GMA. CPC consisted of tricalcium phosphate, monocalcium phosphate monohydrate, dicalcium phosphate dihydrate, and xanthane. Both cements were implanted into cavities drilled in rabbit femoral and tibial condyles. After 2, 4, 12, and 24 weeks of implantation, histological observations and biomechanical tests were performed. With CPC, a progressive osteointegration with a concomitant biodegradation in the presence of macrophages were observed. The mechanical study revealed a decrease of the compressive strength until the 4th week, followed by a slight increase. There was a general decrease in the elastic modulus with time. Moreover, by week 4, the histological study showed that the new bone was in direct contact with CPC margins. No inflammation was observed during the observation period. With PRC, the osteointegration as well as the biodegradation were slight, but its compressive strength was higher than that of cancellous bone and CPC (p < 0.05) at all observation periods. Its elastic modulus was greater than that of cancellous bone and CPC until the 4th week, then fell under the values of the cancellous bone. PMID- 10458274 TI - Use of autologous growth factors in lumbar spinal fusion. AB - The results of spinal fusion, especially posteriorly above the lumbosacral junction, have been mixed. Autologous growth factor concentrate (AGF) prepared by ultraconcentration of platelets contains multiple growth factors having a chemotactic and mitogenic effect on mesenchymal stem cells and osteoblasts and may play a role in initiating bone healing. The purpose of this retrospective study is to review our results with AGF in lumbar spinal fusions. To date, AGF has been used in 39 patients having lumbar spinal fusion. The study group consisted of the first 19 consecutive cases to allow at least 6 months follow-up. The average follow-up was 13 months (range 6 to 18 months). Follow-up compliance was 91%. There were 7 men and 12 women. Average age was 52 years (range 30-72 years). Nine patients had prior back surgery. There were 8 smokers. AGF was used in posterior (n = 15) or anterior intradiscal (n = 4) fusions. AGF was used with autograft and coraline hydroxyapatite in all posterior fusions, and autograft, coral, and intradiscal spacer (carbon fiber spinal fusion cages or Synthes femoral ring) in intradiscal fusions. Posterior stabilization was used in all cases. Eight cases were single-level fusions, 6 were two-level, and 1 was a three level fusion. Autologous iliac crest bone graft was taken in 14 cases and local autograft used in 5 cases. Posteriorly, a total of 23 levels were fused; of these, nine were at L5-S1, eight at L4-L5, five at L3-L4, and one at L2-L3. No impending pseudoarthroses were noted on plain radiographic examination at last follow-up visit. Solid fusion was confirmed in 3 patients having routine hardware removal, and in 2 patients who had surgery at an adjacent level. There was one posterior wound infection, which was managed without sequelae. When used as an adjunct to autograft, AGF offers theoretical advantages that need to be examined in controlled studies. Further study is necessary to determine whether coralline hydroxyapatite used as a bone graft extender in lumbar spinal fusion may help to obviate the need for secondary site graft harvesting. PMID- 10458275 TI - Association of porous hydroxyapatite and bone marrow cells for bone regeneration. AB - The preparation of hybrid material with osteoinductive capacity may be achieved by association of cultured autologous bone cells with a porous ceramic vehicle. We optimized culture conditions for rabbit marrow stromal stem cells (MSCs), notably by selection from batches of fetal calf serum. Rabbit MSCs formed colony forming unit-ribroblastic (CFU-Fs) in vitro. Their alkaline phosphatase (ALP) activity was doubled in the presence of dexamethasone. Autologous rabbit serum allowed the formation of ALP-positive CFU-Fs, but results were highly variable depending on the rabbit. We tested the osteogenic potential of autologous cultured (with or without dexamethasone addition in the culture medium) and noncultured rabbit MSCs associated with a porous hydroxyapatite ceramic after a dorsal intramuscular implantation. Nucleated cells (10(7) or 10(8)/mL) were used for the preparation of autologous hybrid material. A significantly higher number of implants containing bone was obtained with a suspension of 10(7) cells/mL cultured in the presence of 10(-8) M dexamethasone. Some positive implants were also obtained with a suspension of 10(8) noncultured cells/mL. We demonstrated the feasibility of preparing rabbit autologous hybrid materials following a process for controlling culture conditions, cell characterization and cell/material association. PMID- 10458276 TI - Evaluation of human recombinant bone morphogenetic protein-2-loaded tricalcium phosphate implants in rabbits' bone defects. AB - Porous beta-tricalcium phosphate (betaTCP) has osteoconductive properties. The adsorption of human recombinant bone morphogenetic protein-2 (rhBMP-2) onto TCP could realize an osteoinductive bone substitute. We evaluated it on an animal model using dual-energy X-ray absorptiometry (DEXA) and solid-state 31P nuclear magnetic resonance (NMR) spectroscopy. BetaTCP cylinders loaded with rhBMP-2 were implanted into rabbits' femoral condyle bone defects, and betaTCP alone as control into the contralateral femur. We studied two different doses of rhBMP-2 (10 and 40 microg) on two groups of four animals. Evaluation consisted in radiography, histology, and histomorphometry, DEXA, and NMR spectroscopy using an original method of quantification. With both doses of rhBMP-2, we observed on radiographs an increase of trabecular bone around implants. Histology showed resorption of the ceramic, trabecular bone with osteoblasts and osteoid substance around the implants, and colonization inside the porous betaTCP by new bone formed. Histomorphometry showed that the osteoid surface (OS/BS) was greatest with the high dose of rhBMP-2. The difference was slight between the low dose of rhBMP-2 and control. DEXA showed a dose-dependent increase of bone mineral density of rhBMP-2-loaded betaTCP vs. control. NMR spectroscopy confirmed that the amount of new bone formed in betaTCP was greater when betaTCP carried rhBMP 2, and increased with the dose of rhBMP-2 used. We showed that betaTCP was a good matrix for rhBMP-2, which gave it osteoinductive properties in an orthotopic site, in a dose-dependent manner. Thus, such composite biomaterial seems to be of great interest in reconstructive bone surgery. Further studies are needed in clinical practice to determine optimal doses. PMID- 10458277 TI - Biphasic calcium phosphate/hydrosoluble polymer composites: a new concept for bone and dental substitution biomaterials. AB - Calcium phosphate materials have been increasingly employed in orthopedic and dental applications in recent years and are now being developed for use in noninvasive surgery or as carriers for drug delivery systems. We developed an injectable bone substitute (IBS) constituted of biphasic calcium phosphate and a hydrosoluble polymer as a carrier. In vivo biocompatibility and biofunctionality of IBS were tested in rabbits using implants in osseous and nonosseous areas. The results obtained demonstrated that the concept of IBS, a filler without initial mechanical properties but able to be rapidly resorbed and replaced by newly formed bone, can be applied to new surgical applications in orthopedic surgery, maxillofacial surgery, and dentistry for pulp capping and root filling. PMID- 10458278 TI - Influence of sterilization on injectable bone biomaterials. AB - Injectable biomaterials used in bone surgery include acrylic bone cements, calcium phosphate cements, and new composite-type biomaterials with a mineral content and an organic phase dispersed or dissolved in water. Cellulose derivatives, chitosan solutions, alginates, and other polymers are studied as useful modifiers and binding agents in calcium phosphate cements. We have developed proprietary polyester copolymers including lactic acid moieties and present here results concerning the effect of sterilization on the physico chemical properties of derived bone biomaterials. Chitosan solutions show a dramatic decrease in viscosity after 25-kGy gamma sterilization. Aqueous copolylactic solutions also show, by capillary electrophoresis, that hydrolysis occurs to liberate monomers after 25-kGy gamma sterilization. Heat sterilization also degrades chitosan solutions, and ultrafiltration is difficult because of high viscosity. However, apatite-copolylactic solids can be steam sterilized without deterioration. Gelatin has been used as a natural polymer to bind apatite particles. Gel exclusion chromatography reveals crosslinking of the chains by irradiation. Standard acrylic cements contain monomers sterilized by ultrafiltration because they do not tolerate irradiation. We have used ultrafiltration to prepare aqueous copolylactic solutions without polymer hydrolysis. Implantation of calcium phosphate cement modified by copolylactic acid in a rabbit metaphyseal model defect shows progressive substitution of the biomaterial by new bone tissue. At 3 months, a mild inflammatory reaction still remains associated with the continuing resorption of the biomaterial. These results show that interesting biological properties can be obtained with products not sterilized by irradiation. Undoubtedly, many biopolymers are fragile and, like reactive monomers, need to be sterilized by special methods if they are to be used in injectable, liquid form. PMID- 10458279 TI - Injectable bone substitute using a hydrophilic polymer. AB - We studied a new injectable biomaterial for bone and dental surgery consisting of a hydrophilic polymer as matrix and bioactive calcium phosphate (CaP) ceramics as fillers. This material is composed of complex fluids whose flow is determined by the laws of rheology. We investigated the macromolecular effects on this composite in a tube. The stability of the polymer and the mixture is essential to the production of a ready-to-use injectable biomaterial. These flow properties are necessary to obtain CaP bioactivity in a dental canal or bone defect during percutaneous surgery. Macromolecules provide spaces between CaP ceramic granules and facilitate the role of the biological agents of bone substitution. PMID- 10458281 TI - Potentialities of ultrasounds for the nondestructive evaluation of cell adhesion. AB - The aim of this paper is to present the potentialities of ultrasounds to investigate the mechanical properties of a cell/substrate interface. The adhesion process plays a major role in the development of osteoblastic cells on various substrates used in orthopedic applications such as metals, bioceramics, etc. Particularly, cell adherence appears to be a critical factor in the colonization process. High-frequency and low-power ultrasounds seem to be an appropriate tool for a nondestructive evaluation of interface properties. First, we present the results obtained with bulk longitudinal and shear waves under an arbitrary incidence over an aluminum-adhesive interface. This study was performed for an industrial application of bonding. The results clearly show the sensitivity of shear waves for the evaluation of the adhesion quality owing to the shear solicitations at the interface they induce. A model of ultrasound interactions with a boundary subject to varying degrees of adhesion has been developed and compared to the experiments. Second, we investigated osteoblastic cell cultures with a high-frequency acoustic microscope working at 50 MHz. The images obtained in the shear mode reveal a better contrast than those obtained in the longitudinal mode. For the time being, these results are qualitative, and theoretical models have to be developed according to the point of view of biologists. PMID- 10458280 TI - Short-term effects of mineral particle sizes on cellular degradation activity after implantation of injectable calcium phosphate biomaterials and the consequences for bone substitution. AB - This in vivo study investigated the influence of two calcium phosphate particle sizes (40-80 microm and 200-500 microm) on the cellular degradation activity associated with the bone substitution process of two injectable bone substitutes (IBS). The tested biomaterials were obtained by associating a biphasic calcium phosphate (BCP) ceramic mineral phase and a 3% aqueous solution of a cellulosic polymer (hydroxypropylmethylcellulose). Both were injected into osseous defects at the distal end of rabbit femurs for 2- and 3-week periods. Quantitative results for tartrate-resistant acid phosphatase (TRAP) cellular activity, new bone formation, and ceramic resorption were studied for statistical purposes. Positive TRAP-stained degradation cells were significantly more numerous for IBS 40-80 than IBS 200-500, regardless of implantation time. BCP degradation was quite marked during the first 2 weeks for IBS 40-80, and bone colonization occurred more extensively for IBS 40-80 than for IBS 200-500. The resorption-bone substitution process occurred earlier and faster for IBS 40-80 than IBS 200-500. Both tested IBS displayed similar biological efficiency, with conserved in vivo bioactivity and bone-filling ability. Differences in calcium phosphate particle sizes influenced cellular degradation activity and ceramic resorption but were compatible with efficient bone substitution. PMID- 10458282 TI - Raman microspectrometry studies of brushite cement: in vivo evolution in a sheep model. AB - Calcium phosphate hydraulic cements are promising synthetic bone grafting materials. Brushite-based cements were implanted for 6 and 12 months in the distal condyle of sheep femur, and their in vivo evolution was investigated by Raman microspectrometry. This new technique can probe small volumes in the cubic micrometer range. Its resolution allows a very fine analysis of crystalline changes in calcium phosphate mixtures at the microscopic level. First, Raman spectra of pure brushite, monetite, and beta-tricalcium phosphate (beta-TCP) were recorded, in order to set a data base for the basic components of brushite cements. These spectra show significant differences in the vibration mode v1 for the phosphate ion (988 and 878 cm(-1) for brushite, 988 and 900 cm(-1) for monetite, 968 and 948 cm(-1) for beta-TCP). These differences are strong enough as to allow the qualitative and quantitative analysis of these crystalline phases in the cement. Implanted sheep femur samples were harvested after 24 and 52 weeks post-op, and prepared for Raman analysis in the form of 1-mm-thick sections. Implants at 24 weeks show a core of residual cement isolated from the surrounding bone by fibroconnective tissue. No trace of brushite was detected by micro-Raman analysis in this area, but instead, a mixture of beta-TCP and Type-B carbonated apatite, the latter being very close in composition and structure to the mineral fraction of normal bone in the vicinity of the implant. Implants recovered after 52 weeks show a decrease of the bone/residual cement perimeter, whereas new trabeculations are formed in the implanted zone; the small amounts of residual cement still present are substantially transformed into Type-B carbonated apatite containing small amounts of proteins. In the same area, some beta-TCP particles are also detected showing that, contrary to brushite, the excess beta-TCP originally present in the cement is not completely metabolized. In the implanted zone already converted into trabecular bone, Raman microspectrometry shows the characteristic spectrum of normal bone. PMID- 10458283 TI - Pathology findings with acrylic implants. AB - We report the pathological findings in cases of acrylic implants obtained by direct intratumoral injection of polymethyl-methacrylate (PMMA) and N-butyl-cyano acrylate (NBCA). Direct intratumoral injection of acrylic implants was performed for a variety of primary and secondary bone lesions. These types of treatments have been used at our institution in the last 4 years for 40 vertebroplasty (PMMA) procedures and for nine bone lesions of other locations (PMMA, NBCA). Postmortem histology became available for 1 case of PMMA and for 5 cases with NBCA intratumoral acrylic implants. The pathological findings associated with PMMA and NBCA were evaluated and compared. PMMA exhibited a macroscopic and microscopic rim of tumor necrosis, 6 months after implantation. NBCA exhibited compressive effects on the nearby tumor tissue, however, without signs of significant necrosis outside the acrylic tumor cast. Tumor captured inside the acrylic cast showed extensive to near complete necrosis. Acrylic implants may lead to necrosis when injected directly in tumors. The necrotizing effect may extend beyond the limits of an implant in the case of PMMA. Such an extended effect of PMMA, when compared with NBCA, may be due to the variable toxicity of acrylic implants, including the different degrees of the exothermic reaction during polymerization. PMID- 10458284 TI - Stimulation of bone marrow cells and bone formation by nacre: in vivo and in vitro studies. AB - There is frequently a loss of vertebral bone due to disease or aging. Nacre (mother of pearl from the oyster Pinctada maxima) stimulates bone cell differentiation and bone formation in vitro and in vivo. Experimental bone defects were prepared in the vertebrae of sheep and used to test the suitability of nacre as an injectable osteogenic biomaterial for treating vertebral bone loss. Twenty-one cavities were prepared in the first four upper lumbar vertebrae of 11 sheep and filled with nacre powder. The lumbar vertebrae were removed after 1 to 12 weeks, embedded undecalcified in methacrylate, and processed for histological studies. The nacre slowly dissolved and the experimental cavities contained a large active cell population. By 12 weeks, the experimental cavity was occupied by newly matured bone trabeculae in contact with or adjacent to the dissolving nacre. The functional new bone trabeculae were covered with osteoid lined with osteoblasts, indicating continuing bone formation. The in vitro study on rat bone marrow explants cultured with a water-soluble extract of the nacre organic matrix also resulted in the stimulation of osteogenic bone marrow cells with enhanced alkaline phosphatase activity. Thus, both the in vivo and in vitro findings suggest that nacre contains one or more signal molecules capable of activating osteogenic bone marrow cells. PMID- 10458285 TI - Reinforcement of osteosynthesis screws with brushite cement. AB - The fixation of osteosynthesis screws remains a severe problem for fracture repair among osteoporotic patients. Polymethyl-methacrylate (PMMA) is routinely used to improve screw fixation, but this material has well-known drawbacks such as monomer toxicity, exothermic polymerization, and nonresorbability. Calcium phosphate cements have been developed for several years. Among these new bone substitution materials, brushite cements have the advantage of being injectable and resorbable. The aim of this study is to assess the reinforcement of osteosynthesis screws with brushite cement. Polyurethane foams, whose density is close to that of cancellous bone, were used as bone model. A hole was tapped in a foam sample, then brushite cement was injected. Trabecular osteosynthesis screws were inserted. After 24 h of aging in water, the stripping force was measured by a pull-out test. Screws (4.0 and 6.5 mm diameter) and two foam densities (0.14 and 0.28 g/cm3) were compared. Cements with varying solid/liquid ratios and xanthan contents were used in order to obtain the best screw reinforcement. During the pull-out test, the stripping force first increases to a maximum, then drops to a steady-state value until complete screw extraction. Both maximum force and plateau value increase drastically in the presence of cement. The highest stripping force is observed for 6.5-mm screws reinforced with cement in low density foams. In this case, the stripping force is multiplied by 3.3 in the presence of cement. In a second experiment, cements with solid/liquid ratio ranging from 2.0 to 3.5 g/mL were used with 6.5-mm diameter screws. In some compositions, xanthan was added to improve injectability. The best results were obtained with 2.5 g/mL cement containing xanthan and with 3.0 g/mL cements without xanthan. A 0.9-kN maximal stripping force was observed with nonreinforced screws, while 1.9 kN was reached with reinforced screws. These first results are very promising regarding screw reinforcement with brushite cement. However, the polyurethane foam model presents noninterconnected porosity and physiological liquid was not modelized. PMID- 10458286 TI - Influence of polymeric additives on the mechanical properties of alpha-tricalcium phosphate cement. AB - Recently, great attention has been paid to calcium phosphate cements, because of their advantages in comparison with conventional calcium phosphate bioceramics employed for bone repairing, regarding in situ handling, and shaping abilities. Nevertheless, the calcium phosphate cements exhibit relatively low mechanical strength. The aim of this work was the improvement of the compressive strength of alpha-tricalcium phosphate-based cement. The hydraulic setting reaction of this system produces a calcium-deficient hydroxyapatite phase suitable for bone repairing: alpha-Ca3(PO4)2 + H2O --> Ca9(HPO4)(PO4)5OH. Mechanical strength can be improved using technological solutions developed for other applications, such as Portland cement and dual-setting glass-ionomers, by using polymeric additives. The additives used in this work were sodium alginate, sodium polyacrylate, and an in situ polymerization system resulting in a polyacrylamide crosslinked hydrogel. Parameters evaluated were setting time, compressive strength before and after immersion in simulated body fluid, density, porosity, crystalline phases, and microstructure. Sodium alginate and sodium polyacrylate were deleterious to both setting time and mechanical strength. When the in situ polymerization system was added, two setting reactions progressed in parallel: the conventional hydraulic reaction and the copolymerization of acrylamide and crosslinking water-soluble monomers. The initial and final setting times of the "dual-setting" cement were 9 and 35 min, respectively, and they can be regulated varying the initiator, catalyst, and monomers concentrations. The initial compressive strength of the dual-setting cement (6.8 MPa at 0 h, and 15.2 MPa at 24 h) is higher than that of unmodified cement. The major crystalline phase after setting is hydroxyapatite. The dual-setting cement seems to be suitable for clinical applications in bone repairing and remodeling. PMID- 10458287 TI - Nuclear magnetic resonance spectroscopy of bone substitutes. AB - Calcium phosphate bone replacement biomaterials are widely used in different applications. Structure, composition, and organization are, before implantation, analyzed with different methods. Among them, X-ray diffraction is a recognized test. As bioresorption produces more amorphous material, the process is observed and quantified via scanning electron microscopy. Comparatively high-resolution 31P solid-state nuclear magnetic resonance spectroscopy is able to analyze raw ceramics composition and to estimate osteoformation. PMID- 10458288 TI - Biomimetic calcium phosphate coatings on Ti6AI4V: a crystal growth study of octacalcium phosphate and inhibition by Mg2+ and HCO3-. AB - The biomimetic approach for coating metal implants allows the deposition of new calcium phosphate (Ca-P) phases. Films elaborated at physiological conditions might have structures closer to bone mineral than hydroxylapatite (HA) plasma sprayed coatings. In this study, different Ca-P coatings have been deposited through a two-step procedure. After cleaning and etching, Ti6Al4V plates were pretreated by soaking in a simulated body fluid (SBF), i.e., a solution containing inorganic components in concentration more or less similar to body fluids: a thin amorphous carbonated Ca-P layer precipitated on the metal substrate. Second, by soaking these thinly coated metal substrates in another SBF, with different concentrations, the thin amorphous carbonated Ca-P layer led to the fast precipitation of a second and thick Ca-P layer. Different SBF solutions were used in order to investigate the influence of magnesium and carbonate ions. From SBF containing only Ca2+ and HPO4(2-) ions, an octacalcium phosphate layer grew epitaxially on the substrate. When Mg2+ was added into this SBF, the coating was composed of Ca-deficient apatite crystals, while the addition of HCO3- in SBF led to the formation of a B-carbonated apatite layer. Magnesium and carbonate acted as inhibitors of crystal growth. The three phases obtained by our biomimetic process are closer to bone mineral structure than plasma-sprayed HA. Therefore, the obtained results may be particularly relevant for the development of biomimetic Ca-P coatings with optimal bioactivity. PMID- 10458289 TI - Retinal detachment in cytomegalovirus retinitis: should our approach be changed? PMID- 10458290 TI - Surgical repair of cytomegalovirus-related retinal detachment without silicone oil in patients with AIDS. AB - PURPOSE: To analyze visual and anatomic results following surgical repair of cytomegalovirus (CMV)-related retinal detachment (RD) without silicone oil permanent tamponade. METHODS: We analyzed five consecutive patients (six eyes) with acquired immunodeficiency syndrome and CMV-related RD that were repaired with pars plana vitrectomy with peeling of the posterior hyaloid, laser photocoagulation, encircling scleral buckle, and intraocular gas tamponade. RESULTS: Preoperative vision ranged from 20/40 to hand motion. Total retinal reattachment was achieved in five of six eyes (83%). Macular reattachment was achieved in all eyes. Mean postoperative visual acuity was 20/40 (range 20/30 20/60). Mean postoperative follow-up was 12 months (range 7-19 months). All patients in this series presented with low preoperative CD4+ T-lymphocyte counts (mean, 24 cells per microL) and received highly active antiretroviral therapy. One retina (Patient 1) redetached 7 months after initial repair and was successfully reattached without using silicone oil. Postoperatively, visual acuity remains 20/30, and total retinal reattachment has been maintained for 16 months. CONCLUSION: Good anatomic and visual success can be achieved and maintained in CMV-related RD without the use of silicone oil. PMID- 10458291 TI - Retinal detachment associated with macular hole surgery: characteristics, mechanism, and outcomes. AB - OBJECTIVE: To study the frequency, features, and outcomes of retinal detachment (RD) occurring after macular hole surgery. METHODS: Retrospective review of all cases of macular hole surgery. Cases with postoperative RD were identified for study. Outcome measures included baseline demographic and ocular characteristics, RD features, surgical procedure, macular hole status, and final visual and anatomic outcomes. RESULTS: Retinal detachment occurred in 8 (1.8%) of 438 eyes undergoing macular hole surgery. The rate of RD was 3.5% early in the course of the surgeon's experience (first 200 cases) and 0.4% later in the surgeon's experience (after 200 cases) (P = 0.026). Two of the eight eyes with RD had undergone previous macular hole surgery. All six primary cases occurred after repair of stage 3 macular holes, which had involved peeling of the posterior cortical face. The RD involved the inferior quadrants in seven eyes and the macula in two eyes. The pathogenic retinal break was in the equatorial region in four eyes, was anterior to the equator in two eyes, and remained undetermined in two eyes. The retina was attached and the macular hole was closed in all cases at the final follow-up examination (mean 30 months). The macular hole reopened 2 years following successful reattachment of the RD in one eye. Final visual acuity was > or =20/60 in four eyes and <20/200 in one eye. CONCLUSION: Retinal detachment after macular hole surgery is uncommon, and may be related to posterior cortical vitreous stripping. Early detection of RD minimizes adverse visual and anatomic outcomes. PMID- 10458292 TI - Corneal epithelial defects following vitrectomy surgery using hand-held, sew-on, and noncontact viewing lenses. AB - PURPOSE: To compare the incidence of corneal epithelial defects following vitrectomy surgery with Charles hand-held infusion lenses, Landers sew-on lenses, and the Oculus BIOM noncontact lens system. METHODS: We performed a retrospective chart review of 234 patients who underwent initial vitrectomy surgery by one surgeon. We determined the presence and duration of postoperative epithelial defects and identified predisposing factors. RESULTS: There were more corneal epithelial defects noted postoperatively in eyes with hand-held infusion lenses (23.8%) compared with eyes with sew-on lenses (8.6%; P = 0.010) and eyes with noncontact lenses (0%; P < 0.001). There were also more defects in sew-on versus noncontact lenses (P = 0.014). Diabetic patients had more postoperative epithelial defects with hand-held infusion lenses (32.1 %) than with sew-on lenses (8.8%; P = 0.011) or with noncontact lenses (0%; P < 0.001). The average operative time for patients with epithelial defects (169 minutes) was longer than for patients without defects (117 minutes) (P < 0.001). The risk of these defects remained significant after controlling for both length of surgery and diabetes status. CONCLUSIONS: Corneal epithelial defects were more common after vitrectomy surgery using hand-held infusion lenses than after surgery using sew-on lenses and did not occur with the noncontact lens system. In addition, reducing the operative time may independently reduce the risk of epithelial defects, regardless of lens type. PMID- 10458293 TI - Sub-Tenon's anesthesia in vitreoretinal surgery: a needleless technique. AB - PURPOSE: To evaluate the safety and efficacy of "needleless" regional anesthesia via the sub-Tenon's route without adjunct facial nerve or subconjunctival block for vitreoretinal surgery. METHODS: Forty-eight consecutive patients were included and studied prospectively. The number, time, and volume of anesthetic solutions given were recorded. Any complications such as severe intraocular pressure rise or orbital hemorrhage were noted. Within 24 hours after surgery, the patients were asked to grade their level of pain during the procedure using a standard 10-point visual analog scale. The use of postoperative analgesic agents was recorded from the nurses' notes. Patient demography, number of previous vitreoretinal operations, preoperative diagnosis, and operative procedures performed were recorded. RESULTS: Thirty-seven (77%) patients needed only one or two infiltrations of anesthetic solution (range 1-5, mean 2.1). Ninety-two percent of patients reported a pain score of 0-2 on the visual analog scale (range 0-5, mean 1.17). Ninety percent of patients required no analgesic or just acetaminophen for the control of postoperative pain. The other patients received a single dose or multiple doses of codeine. All patients had adequate akinesia and anesthesia. No complications were encountered. CONCLUSION: Needleless regional anesthesia appears to be safe and effective and can thus be considered as an alternative method of anesthesia in suitable vitreoretinal cases. PMID- 10458294 TI - Focal retinal pigment epithelial dysplasia associated with fundus flavimaculatus. AB - BACKGROUND: One or more focal dysplastic lesions of the retinal pigment epithelium (RPE) occurred in 15 eyes of 10 patients with fundus flavimaculatus. METHODS: Review of patient records including an attempt to obtain follow-up information concerning a history of previous ocular trauma. RESULTS: Mild antecedent ocular trauma occurred to the eye with a dysplastic lesion in two patients. Dysplastic lesions were most frequently solitary and located temporal to the macula. Subretinal neovascularization accompanied two of the dysplastic lesions. The lesions were multifocal and present bilaterally in two patients. CONCLUSIONS: In fundus flavimaculatus, progressive lipofuscin storage is responsible for engorgement and hypertrophy of the RPE. Dysplastic lesions of the RPE probably result from reactive hyperplasia and fibrous metaplasia of RPE cells in response to acute disruption of fragile, hypertrophied RPE cells that may be enormously enlarged in the area of yellow flecks. This disruption may occur in response to trauma, focal inflammation, or other localized stimuli. Patients with fundus flavimaculatus should be cautioned concerning the possible role of trauma in causing dysplastic changes in the RPE and visual loss. PMID- 10458295 TI - Fluorescein angiography of extreme peripheral retina and rubeosis iridis in proliferative diabetic retinopathy. AB - PURPOSE: To study the fluorescein angiographic appearance of the extreme peripheral retina in patients with proliferative diabetic retinopathy with rubeosis iridis. METHODS: Using a modified endoscope, fluorescein angiography was performed prospectively during pars plana vitrectomy on 14 eyes of 12 patients with proliferative diabetic retinopathy who had or developed postoperative rubeosis. These findings were compared with those obtained from 41 eyes of 35 patients without rubeosis. RESULTS: A fibrovascular ridge was observed at the ora serrata in 11 of 12 eyes (92%) with rubeosis but in only 15 of 35 (43%) eyes without rubeosis. Seven of 12 (58%) eyes with rubeosis showed the continuous type of fibrovascular ridge, whereas only 5 of 35 (14%) eyes without rubeosis showed the continuous type. A broom-shaped continuous fibrovascular ridge was observed only in eyes with rubeosis. CONCLUSIONS: Many eyes with proliferative diabetic retinopathy showed unusual findings in the extreme retinal periphery, which were more frequent in the eyes with rubeosis iridis. Fluorescein angiography of this region may provide valuable information for possible anterior neovascularization. PMID- 10458296 TI - Acute posterior multifocal placoid pigment epitheliopathy associated with Wegener's granulomatosis. AB - PURPOSE: To report acute posterior multifocal placoid pigment epitheliopathy (APMPPE) in the course of systemic Wegener's granulomatosis. METHODS: A complete ophthalmologic evaluation was undertaken, including fluorescein and indocyanine green angiography. RESULTS: Two patients diagnosed with Wegener's granulomatosis developed multiple white-yellow subretinal areas with smooth margins typical of APMPPE. Case 1 presented with bilateral choroidal involvement and a granulomatous anterior uveitis. In Case 2, placoid lesions were diagnosed 2 months after onset of systemic Wegener's granulomatosis. In both cases, fluorescein angiography showed an early hypofluorescence and a late staining of the lesions. Indocyanine green angiography revealed early and late hypofluorescence corresponding to the areas seen clinically. Wegener's granulomatosis was diagnosed histologically in both cases. After combined steroid and cyclophosphamide therapy, the lesions healed, and after a follow-up period of 1.5 and 4 years, respectively, funduscopic and angiographic examinations showed cicatricial lesions. CONCLUSION: Acute posterior multifocal placoid pigment epitheliopathy is a rare manifestation of Wegener's granulomatosis and should be considered a possible systemic involvement of the disease. A prompt, thorough investigation should be undertaken for an underlying systemic disorder. PMID- 10458297 TI - Eye injuries caused by cow horns. AB - PURPOSE: To assess ocular injuries caused by cow horns; to investigate clinical findings, treatment, and visual outcome in a population of dairy farmers; and to propose possible preventive measures. METHODS: A retrospective review was conducted to identify patients seen over a 45-month period with cow horn inflicted eye injuries. Eleven patients were identified and their charts reviewed for demographics, mechanism of injury, initial and final visual acuity, surgeries performed, and anatomic outcome. RESULTS: The mean age of the patients was 64 years. Seven patients had open-globe injuries with vitreous hemorrhage. In five cases, pars plana vitrectomy was performed. Final best-corrected visual acuity was < or = hand motion in five patients, between 20/160 and 20/80 in three patients, and better than 20/32 in three patients. CONCLUSION: The majority of the cow horn injuries studied caused severe permanent impairment of vision. Owing to the blunt nature of the horns, a significant amount of energy is imparted into the eye. To prevent these injuries, coagulation of the horns should be performed 2 weeks after a calf's birth or farmers should be advised to wear safety glasses. PMID- 10458298 TI - Use of microdissection and molecular genetics in the pathologic diagnosis of retinoblastoma. AB - BACKGROUND/PURPOSE: Retinoblastoma results from mutations or loss of both alleles of the retinoblastoma gene. Although retinoblastoma is usually recognized clinically, some forms of the disease can elude diagnosis. The purpose of this study was to determine whether the use of molecular genetics to detect a loss of heterozygosity (LOH) in the retinoblastoma gene could assist the ocular pathologist in the diagnosis of this malignancy. METHODS: Deoxyribonucleic acid (DNA) was obtained from tumor cells microdissected from three ocular specimens from two patients with diffuse retinoblastoma. Polymerase chain reaction was used to detect two microsatellite markers (D13S153 and D13S118) of the retinoblastoma gene. Loss of heterozygosity was identified when one of the two polymorphic alleles was present in the DNA from normal tissue but absent or reduced in the DNA obtained from tumor cells. RESULTS: Loss of heterozygosity was identified in all three specimens from the two patients with diffuse retinoblastoma. In one patient, the diagnosis of retinoblastoma was based on identification of LOH from tumor cells obtained from vitrectomy. CONCLUSIONS: This study demonstrates that identification of LOH in retinoblastoma cells not only can contribute to our understanding of the molecular genetics of this tumor, but also can help the ocular pathologist in the diagnosis of atypical forms of the disease. PMID- 10458299 TI - Treatment of herpes retinitis in an animal model with a sustained delivery antiviral drug, liposomal 1-O-octadecyl-SN-glycerol-3-phosphonoformate. AB - PURPOSE: To evaluate the clinical treatment efficacy of a long-lasting intravitreous injectable anti-cytomegalovirus (CMV) liposomal drug, 1-O-octadecyl sn-glycerol-3-phosphonoformate (ODG-PFA). METHODS: Sixty-four pigmented rabbits were used for evaluation of the potency and duration of action of ODG-PFA after intravitreal injection using a herpes simplex virus (HSV)-1 retinitis model. For the potency evaluation, liposomal ODG-PFA was injected into rabbit vitreous at the same time that HSV-1 virus was inoculated onto the retina (simultaneous treatment). For the duration evaluation, ODG-PFA was injected days or weeks before inoculation (pretreatment). Retinitis was clinically graded by indirect ophthalmoscopy, and the retinitis scores were compared across the treatment and control groups. RESULTS: Simultaneous treatment study revealed that ODG-PFA was much more potent than its parent compound, foscarnet (P = 0.0027). Pretreatment study indicated that ODG-PFA possesses a much longer antiviral effect (at least 2 weeks) than foscarnet after a single intravitreal injection. CONCLUSION: Liposomal ODG-PFA is a potent long-lasting intravitreal injectable antiviral compound that may be an ideal alternative for treatment of CMV retinitis in patients with acquired immunodeficiency syndrome. PMID- 10458300 TI - Histopathologic study of presumed parafoveal telangiectasis. AB - PURPOSE: To report the postmortem histopathologic features that closely resemble the clinical features of parafoveal telangiectasis. METHODS: Light and electron microscopy. RESULTS: Histopathologic features included macular edema; telangiectatic vessels; retinal, subretinal, and superficial retinal neovascularization; retinal pigment epithelial hyperplasia around neovascular aggregates; retinal-choroidal vascular anastomosis; and superficial pigmented cells with lipofuscin. CONCLUSION: The postmortem histopathologic findings in a 36-year-old woman with Down syndrome and other systemic conditions correlate with features noted in previous reports of presumed parafoveal telangiectasis. PMID- 10458301 TI - Diagnostic and therapeutic challenges. PMID- 10458302 TI - Giant retinal tears after photorefractive keratectomy. PMID- 10458303 TI - Uveal non-Hodgkin's lymphoma with epibulbar extension simulating choroidal effusion syndrome. PMID- 10458304 TI - Choroidal malignant melanoma associated with a melanocytoma. PMID- 10458305 TI - Choroidal leiomyoma: report of a case. PMID- 10458306 TI - Lecythophora mutabilis endophthalmitis after long-term corneal cyanoacrylate. PMID- 10458307 TI - Positive vitreous cultures in eyes with retained lens fragments. PMID- 10458308 TI - Bilateral Bartonella-associated neuroretinitis. PMID- 10458309 TI - Coats' response in Leber's congenital amaurosis. PMID- 10458310 TI - Progressive presumed choriocapillaris atrophy after surgery for ARMD. PMID- 10458311 TI - Effects of the Resident Assessment Instrument on the care process and health outcomes in nursing homes. A review of the literature. AB - The objective of the paper is to review the effects of the implementation of the Resident Assessment Instrument (RAI) on process measures (quality of care plans and staff satisfaction) and outcome measures (health problems and quality of life) in nursing homes. All available publications on the effects of the RAI were included in the review. The most positive effects of the RAI were found in improvements in the comprehensiveness and accuracy of the care plans. As regards outcome quality, the RAI method had most positive effects on the health condition of nursing home residents with diminished physical and mental functioning. In psychosocial areas of assessment, fewer positive effects were found. We concluded that positive effects have been found, based on pre-test-post-test noncontrolled designs. Control-group designs are needed in future evaluation studies to determine if these positive results will hold. PMID- 10458312 TI - A critical analysis of randomised clinical trials on neck pain and treatment efficacy. A review of the literature. AB - The efficacy of physiotherapy or chiropractic treatment for patients with neck pain was analysed by reviewing 27 randomised clinical trials published 196-1995. Three different methods were employed: systematic analyses of; methodological quality; comparison of effect size; analysis of inclusion criteria, intervention and outcome according to The Disablement Process model. The quality of most of the studies was low; only one-third scored 50 or more of a possible 100 points. Positive outcomes were noted for 18 of the investigations, and the methodological quality was high in studies using electromagnetic therapy, manipulation, or active physiotherapy. High methodological quality was also noted in studies with traction and acupuncture, however, the interventions had either no effect or a negative effect on outcome. Pooling data and calculation of effect size showed that treatments used in the studies were effective for pain, range of motion, and activities of daily living. Inclusion criteria, intervention, and outcome were based on impairment in most of the analysed investigations. Broader outcome assessments probably would have revealed relationships between treatment effect and impairment, functional limitation and disability. PMID- 10458313 TI - A narrative review on classification of pain conditions of the upper extremities. AB - Local and regional musculoskeletal discomfort and pain in the shoulder girdle or upper extremities are often reported, especially in the working population. In this review we describe the most important problems and factors when classifying musculotendinous pain in the upper extremities and shoulders. This includes an analysis of how four common diagnoses (wrist tenosynovitis, lateral epicondylitis, rotator-cuff tendinitis, myofascial pain syndrome) fulfil basic criteria of validity. It is evident that there are some serious problems regarding the validity of the current classification of the conditions. Clinical criteria are often poorly defined and the reliability insufficiently tested. The relationship to objective pathoanatomic or physiological findings seems inconsistent. Although magnetic resonance and ultrasonographic imaging are promising, they are still only preliminary methods for evaluation of tendon and connective tissue structures. The prognosis with and without treatment also seems heterogeneous and can vary between studies. A generally accepted terminology is lacking in the pathogenetically complex regional muscle pain conditions. PMID- 10458314 TI - Functional magnetic resonance imaging of the human motor cortex before and after whole-hand afferent electrical stimulation. AB - Electrical stimulation of the whole hand using a mesh-glove has been shown to improve volitional movement of the hand and arm, and decrease muscle hypertonia after hemispherical stroke in patients who have reached a recovery plateau. The goal of this study was to investigate the effect of stimulation of the nerve afferents of the hand on brain cortical activity elicited by whole-hand subthreshold stimulation for sensation in humans with intact nervous systems. Brain cortical activity in 6 healthy subjects (30-45 years) was studied using blood oxygenation level-dependent functional Magnetic Resonance Imaging during a test motor task, finger-to-thumb tapping and after 20 minutes of mesh-glove stimulation of the resting hand prior to performance of an identical motor task, to test the changes in the conditioned motor task established after 20 minutes of mesh-glove stimulation. Fifteen contiguous echo-planar sequences parallel to the bicommissural plane were acquired for functional magnetic resonance. Post processing of image data included correction of motion artefacts and calculation of correlation coefficients between the signal intensity of pixels during rest and finger tapping and a rectangular reference wave function. The functional Magnetic Resonance Imaging examinations revealed a signal increase in the primary and secondary motor and somatosensory areas when comparing the number of activated pixels during test and conditioned motor tasks. Our preliminary study indicated that change occurred in a definite pattern in the region of the regional cerebral blood flow of the brain cortex after mesh-glove whole-hand stimulation at the subthreshold level for sensation. We assumed that this increase in regional cerebral blood flow also reflected augmented neuronal activity. PMID- 10458315 TI - The influence of non-paretic leg movement on muscle action in the paretic leg of hemiplegic patients. AB - The aim of this investigation was to study the effect of non-involved-side exercise on the involved (paretic) leg-muscle of patients with stroke. Each of the ten adults studied attempted to perform, while seated, a predetermined sequence of the following actions : (1) a voluntary knee extension on the involved side; (2) a maximal, isometric knee-flexion on the non-involved side; (3) a maximal, isometric knee-flexion on the non-involved side with counter resistance (in front of the ankle joint) on the involved side. The root-mean square voltage was measured across the rectus femoris, the medial hamstring, the tibialis anterior, and the medial gastrocnemius muscles (on the involved side) with surface electrodes. The root-mean-square voltages of the involved-side rectus femoris and the tibialis anterior muscles were found to increase substantially during non-involved-side knee flexion relative to that detected for voluntary knee extension on the involved side. The voltage of the rectus femoris muscle increased substantially also when counter-resistance was present. There was no similar substantial increase of muscular activity, in the medial hamstring and medial gastrocnemius, induced by contralateral isometric knee-flexion. This study suggests that, among patients with severe hemiplegia, isometric knee flexion in the non-involved side may be useful for facilitating the paretic rectus femoris and tibialis anterior muscular activities. PMID- 10458316 TI - Muscular dystrophy in adults: a five-year follow-up. AB - The aim was to describe the natural history of adults with hereditary muscular dystrophies, including myotonic dystrophy, with respect to muscular function, ventilation and electrocardiogram. In a prospective study, 46 subjects were followed over a period of five years. In 1991 and 1996, their muscle function was assessed according to an observation scheme and their lung vital capacity was measured by spirometer. Electrocardiograms were obtained in 1991, 1993 and 1996. Deterioration of muscular function was seen with regard to both the functional muscle tests and the vital capacity. The proportion of pathological electrocardiograms increased from 38% in 1991 to 54% in 1996 in the 26 patients with myotonic dystrophy without an increase in clinically detected cardiac abnormalities. Timely examinations using standard methods can reveal medically important information on deterioration, which often passes clinically unnoticed because of the insidious progress of the diseases. PMID- 10458317 TI - Plantar force distribution in Parkinsonian gait: a comparison between patients and age-matched control subjects. AB - This study aimed to ascertain whether roll-off of the feet during gait was essentially different in patients with Parkinson's disease from that of elderly control subjects. Twenty-two patients, belonging mainly to Hoehn & Yahr grades III and IV, and 30 elderly people participated in the study. Plantar force distribution data were collected of two consecutive strides using pressure sensitive insoles as part of the pododynograph system. Results showed that when correcting for gait speed and sex differences, patients with Parkinson's disease walked with significantly lower relative peak forces at the forefoot and heel and increased load at the midfoot. The onset of peak forces indicated slower load acceptance on the heel and early forefoot loading which was confirmed by a reduced amplitude of the centre of force along the length of the foot compared with healthy controls. Roll-off was significantly reduced in patients with Parkinson, a feature which was specific for the disease rather than a result of reduced gait speed alone. PMID- 10458318 TI - The CBF.78 monoclonal antibody to human sialophorin has distinct properties giving new insights into the CD43 marker and its activation pathway. AB - We confirm here the CD43 specificity of the CBF.78 monoclonal antibody (mAb) and compare its phenotypic and functional capacities to classical group-A mAbs (DFT1, MEM-59) and to 2 other CD43 mAbs (RDP/AD9, 161-46). It reacts with stable human CD43 transfectants in a sialic acid independent way and blocks completely cell binding of RDP/AD9 or 161-46 more or less but not DFT1 and MEM-59. Its distribution differs from all other CD43. B lymphocytes, but surprisingly the majority of granulocytes or monocytes are CBF.78 negative. CBF.78 is expressed on all T lymphocytes, but the number of CBF.78 molecules/cell is low and equally represented on resting T CD4 and CD8 cells. In comparison to naive T lymphocytes, CD45RO cells increase their CBF.78 epitopes much more than other CD43 epitopes. At a single cell level, confocal microscopy shows that CBF.78 can exist independently of other epitopes. CBF.78 is able to induce homotypic adhesion in different cell lines but not in peripheral blood lymphocytes and is unable to relocalise the targeted molecules. U937 cell line that is not agglutinated by CBF.78 (or RDP/AD9) undergoes a stronger adhesion with PMA, when this reagent is combined with this mAb. By itself CBF.78 is unable to activate T lymphocytes and to costimulate CD3 mAbs but partially blocks PMA. The phosphorylation of the tyrosine kinase p59fyn and p56lck, driven by CBF.78, is weak and almost blocked by PMA. Altogether these data support the hypothesis that there are at least 3 modes of interaction between PKC and CD43 pathways: each pathway is inhibitory towards the other but the CD43 one can also be synergistic. PMID- 10458319 TI - Relevance of individual CD5 extracellular domains on antibody recognition, glycosylation and co-mitogenic signalling. AB - CD5 is a type I glycoprotein which modulates T- and B-cell receptor-mediated signals and is expressed by thymocytes, mature T cells and a subset of mature B cells. The extracellular region of CD5 is composed of three scavenger receptor cysteine-rich domains (D1, D2, D3) for which only limited functional and structural data are available. Using cell transfectants expressing ectodomain deficient CD5 molecules or CD5 immunoglobulin fusion proteins, we analysed individual CD5 domains with respect to monoclonal antibody binding specificity, glycosylation, and co-mitogenic signalling. Our results show the presence of N linked oligosaccharides on D1 and D2, but not on D3. D1, the most amino-terminal domain, is predicted to be the most appropriately placed domain for an interaction with a ligand. This domain is recognised by a large panel of well characterised CD5 mAbs, reflecting its higher immunogenicity. In an attempt to develop mAbs with specificity for the more conserved membrane-proximal domains, we generated a unique mAb, named 83-C4, whose binding mapped to D3. Co stimulatory studies revealed no significant differences between anti-D1 and anti D3 mAbs. The high interspecies conservation of D3 implies a conserved role of this domain in CD5 function and the 83-C4 mAb promises to be a valuable tool in exploring this. PMID- 10458320 TI - Molecular characterization of a novel human natural killer cell receptor homologous to mouse 2B4. AB - Natural killer (NK) cells spontaneously detect and kill cancerous and virally infected cells through receptors that transduce either activating or inhibiting signals. The majority of well studied NK receptors are involved in inhibitory signaling. However, we have previously described an activating receptor, 2B4, expressed on all murine NK cells and a subset of T cells that mediate non-major histocompatibility complex (MHC) restricted killing. Anti-2B4 monoclonal antibodies directed against IL-2-activated NK cells enhanced their destruction of tumor cells. Recently, we determined binding of 2B4 to CD48 with a much higher affinity than CD2 to CD48. Here we describe the molecular characterization of a cDNA clone homologous to mouse 2B4, isolated from a human NK cell library. The cDNA clone contained an open reading frame encoding a polypeptide chain of 365 amino acid residues. The predicted protein sequence showed 70% similarity to murine 2B4. Additionally, it has 48, 45, and 43% similarity to human CD84, CDw150 (SLAM), and CD48, respectively. RNA blot analysis indicates the presence of 3 kb and 5 kb transcripts in T- and NK-cell lines. A single transcript of 3 kb is identified in poly(A)+ RNA from human spleen, peripheral blood leukocytes, and lymph node, whereas, the level of expression in bone marrow and fetal liver was indeterminate. Preliminary functional data suggests that NK-cell interaction with target cells via 2B4 modulates human NK-cell cytolytic activity. PMID- 10458321 TI - Molecular analysis of HLA class I alleles in the Mexican Seri Indians: implications for their origin. AB - The molecular analysis of HLA class I loci has demonstrated that, although, the genetic profile is restricted in Amerindians, several micropolymorphisms may be important in conferring a biological advantage. We analyzed the HLA-A and B genetic profile of Seris, a Mexican Indian tribe living in northwestern Mexico in the state of Sonora. There are presently only 619 individuals. Our study included 100 Seris belonging to nine families. HLA-A and -B loci typing was performed by polymerase chain reaction using an amplification refractory mutation system (PCR ARMS) on a select group of samples; all of them were typed by polymerase chain reaction using sequence-specific oliogonuoleotide probes (PCR-SSOP) at a low intermediate resolution level. The correlation between the techniques was 100%. Only five HLA-A alleles and seven HLA-B alleles were found. A*0201, A*68, A*31, A*24, B*3501, B*40, B*51, B*3512 and B*15 were present in over 5% of the individuals. B*27052 was detected in 2%. B27 is absent in any other Mexican Indian groups previously studied. The presence of B27 may be the result of a founder effect due to different waves of southward migrations. The B-locus is more diverse and the prevalent haplotypes were: A*0201-B*3501, A*0201-B*40, A*0201-B*3512, A*31-B*51, A*68-B*3501 and A*68-B*40. This genetic profile is different from the pattern of other Mexicans. The phylogenetic tree suggests that Seris are more closely related to the Warao Indians from Venezuela, who live in a similar ecosystem, and to some groups of Argentina, than they are to the Mexican Lacandones who live in the jungle. These data emphasize the relevance of the interaction between genes and environment. PMID- 10458323 TI - HLA genotyping of 5,000- and 6,000-year-old ancient bones in Japan. AB - We used polymerase chain reaction (PCR)-based DNA typing to identify HLA class II alleles of two individuals from ancient human remains. Genomic DNAs were isolated from two ancient human skeletons excavated from the Sanganji and Kitakogane sites in the main and northern islands of Japan, respectively. They were archaeologically estimated to be approximately 5,000 and 6,000 years old respectively, representing the remnants from the Jomon era. High molecular weight DNA was extracted by the standard proteinase K-phenol extraction method followed by purification with a Centricon-30 micro concentrator. Several rounds of PCR successfully gave rise to amplification of the HLA-DRB1 and -DQA1 genes. The PCR restriction fragment length polymorphism (PCR-RFLP) and sequencing based typing (PCR-SBT) methods revealed that those ancient individuals possessed the DRB1 and DQA1 alleles which are highly prevalent among the modern north Asian as well as Japanese populations. PMID- 10458322 TI - An allelic non-histocompatibility antigen with wide tissue distribution as a marker for chimerism in pigs. AB - It is frequently useful in studies of transplantation to have available an antibody to a cell surface antigen, which is not itself responsible for transplant rejection. In this paper, we identify and describe such an antibody/antigen system in miniature swine. The monoclonal antibody, 1038H-10-9, was found to react to a pig allelic antigen (called PAA), found on a variety of pig cells and tissues, including peripheral blood mononuclear cells (PBMC), thymocytes, lymph node, bone marrow, and skin. Analysis for recipient sensitization against PAA was performed by in vitro cell-mediated lympholysis (CML) assay, mixed lymphocyte reaction (MLR) assay, antibody binding studies, and skin graft rejection patterns were examined. No evidence was found to indicate detection of PAA by any of these assays of alloreactivity. We therefore conclude that PAA is an allelic swine cell surface antigen, with wide tissue distribution, and that it is not a histocompatibility antigen. It should provide a powerful tool for studies of transplantation biology in miniature swine, such as identification and quantification of chimerism following organ transplantation. PMID- 10458324 TI - Allele resolution of HLA-A using oligonucleotide probes in a two-stage typing strategy. AB - High-resolution polymerase chain reaction using sequence-specific oligonucleotide probes (PCR-SSOP) typing methods for HLA-A identification have been established. The four systems, which operate independently of each other, are intended for use as secondary typing systems following HLA-A identification with a medium resolution PCR-SSOP technique. The systems, all using digoxigenin-labelled probes, are based on group specific amplifications for resolution of: i) HLA-A*29 & -A*33; ii) HLA-A*24 & -A*30; and iii) HLA-A*26, -A*25, -A*11, -A*34, -A*66 and A*68 alleles, respectively. The fourth system, for the detection of HLA-A*02 alleles, is a modification of a previously reported PCR-SSOP subtyping system. The methods have been applied to individuals from the local bone marrow registry and HLA-A allele frequencies for the Northern Ireland population have been established. PMID- 10458325 TI - High-resolution sequence typing of HLA-DQA1 and -DQB1 exon 2 DNA with taxonomy based sequence analysis (TBSA) allele assignment. AB - High-resolution DNA sequencing of exon 2 of DQA1 and DQB1 genes that uses a taxonomy-based sequence analysis (TBSA) method to assign alleles was developed. The system uses fewer primers for polymerase chain reaction (PCR) amplification and sequencing than other methods and yields accurate DQA1 and DQB1 typing when either homozygous or heterozygous DNA samples are tested. The approach was initially corroborated by the correct typing of 10 blinded samples that had been previously typed by PCR using sequence-specific oligonucleotide probes (PCR-SSOP) or serology, and subsequently confirmed by sequencing of cloned PCR products. DNA from peripheral blood cell samples of 130 individuals enrolled in a case-control analysis of HLA determinants of abdominal aortic aneurysm were subsequently evaluated. Overall, 8 different DQA1 and 19 DQB1 alleles were identified. All 21 DQA1 heterozygous combinations and 45 of 49 DQB1 heterozygous combinations were successfully resolved with TBSA. The two pairs of heterozygous DQB1 combinations that were not unambiguously typed required sequence specific PCR amplification for correct allele identification. We conclude that the method provides precise analysis for HLA-DQ typing. PMID- 10458326 TI - Association of HLA class II alleles with different subgroups of diabetes mellitus in Eastern India identify different associations with IDDM and malnutrition related diabetes. AB - Genetic studies of Malnutrition related diabetes are few. We have analyzed HLA class II gene polymorphism in different types of diabetes mellitus patients from Cuttack in Eastern India. Patients with insulin-dependent diabetes mellitus (IDDM), non-insulin-dependent diabetes mellitus (NIDDM) and malnutrition-related diabetes mellitus (MRDM), which is subdivided into protein-deficient diabetes mellitus (PDDM) and fibrocalculous pancreatic diabetes (FCPD), were studied and their associations with autoantibody markers. IDDM and PDDM were associated with DR3 and DQ2 but not DR4 and DQ8. FCPD was positively associated with DQ9 (A*0201 B*0303). The association of DQ9 with FCPD suggests differences in the genetic background for susceptibility between IDDM and MRDM in the Cuttack population. There is no association seen between HLA-DR-DQ and NIDDM patients from Eastern India. Clinical classification of diabetes into IDDM, NIDDM and MRDM does not identify the underlying pathological mechanisms. Presence of autoantibodies to IDDM autoantigens in clinical MRDM and NIDDM identifies the slow-onset form of IDDM. Due to the absence of autoantibody assays for diagnosis of IDDM in India, slow onset IDDM is not diagnosed and the patients are classified as NIDDM or MRDM. Our study demonstrates that the presence of GAD65 antibody and DR3-DQ2 positivity in MRDM and NIDDM patients in Eastern India would suggest the presence of slow-onset IDDM. Our data would indicate alternatively that MRDM can coexist with IDDM in these patients and malnutrition could be one of the reasons for the slower onset in IDDM-prone individuals. PMID- 10458327 TI - HLA-DRB1*0403 is associated with dominant protection against IDDM in the general Dutch population and subjects with high-risk DQA1*0301-DQB1*0302/DQA1*0501 DQB1*0201 genotype. AB - Insulin-dependent (Type 1) diabetes mellitus (IDDM) is a genetically controlled T cell mediated autoimmune disease. Recently, subtyping of HLA-DRB1*04 identified the HLA-DRB1*0403 allele to be associated with protection in Caucasoids with the highest risk heterozygous genotype DQA1*0301-DQB1*0302/DQA1*0501-DQB1*0201. Some studies confirmed this finding, but other reports were not consistent with a dominantly protective trait. We here report the frequency of HLA-DRB1*0403 in a large cohort (n=200) of Dutch patients with IDDM, their first-degree family members (n=370), and random controls (n=420) of the general population in The Netherlands. We found that HLA-DRB1*0403 is strongly associated with dominant protection against development of IDDM in unrelated subject, even in the context of the highest risk HLA-DQ phenotypes and HLA-DR4-DQB1*0302 (P < 0.0001). PMID- 10458328 TI - Histocompatibility antigens and alleles in Japanese haemophilia A patients with or without factor VIII antibodies. AB - We carried out human leukocyte antigen (HLA)-A, B, Cw, DR and DQ serological typing and HLA-DQA1, DQB1, DRB1 and DPB1 genetic typing for 46 Japanese haemophilia A patients, including 20 who had developed an antibody to factor VIII. It appears that anti FVIII inhibitor formation is associated with the major histocompatibility complex in Japanese haemophilia A patients. Absence of HLA-A24 is a principal risk factor for inhibitor formation in Japanese haemophilia A patients. As supplemental risk factors, HLA-DR4.1, DQ4 and DQA1*0301=2 are positively associated with patients exhibiting inhibitor compared with normal subjects. This and previous studies show that the association between HLA antigens and the formation of inhibitor depends on race. Data of HLA typing may be useful for the recognition of groups at high risk for the possible formation of inhibitor among Japanese haemophilia A patients. PMID- 10458329 TI - Characterization of a novel HLA-A*24 allele containing an HLA-A*03 sequence motif. AB - HLA-A*2418 is described for the first time. It segregates in a Mendelian fashion. Serological analyses indicate that the new allele encodes epitopes from both HLA A9 and -A3 specificities. Results from nucleotide sequencing analyses of polymerase chain reaction (PCR) amplification products derived from genomic and cDNA are compatible with those findings. PMID- 10458330 TI - Rose Payne 1909-1999. With personal recollections by Julia and Walter Bodmer. PMID- 10458331 TI - Nomenclature for factors of the HLA system, update January 1999. Marsh for the WHO Nomenclature Committee for Factors of the HLA System. PMID- 10458333 TI - Nomenclature for factors of the HLA system, update March 1999. Marsh for the WHO Nomenclature Committee for Factors of the HLA System. PMID- 10458332 TI - Nomenclature for factors of the HLA system, update February 1999. Marsh for the WHO Nomenclature Committee for Factors of the HLA System. PMID- 10458334 TI - The use of hyperbaric oxygen in urology. AB - PURPOSE: We review the use of hyperbaric oxygen therapy in urology, and present the mechanisms of hyperoxia action in whole body hyperbaric chamber treatments, patient outcomes and patient selection criteria. MATERIALS AND METHODS: The literature on hyperbaric oxygen use in urology was reviewed. RESULTS: Hyperbaric oxygen is a treatment alternative for patients with an underlying ischemic process unresponsive to conventional therapy. Specific factors which may influence patient selection of hyperbaric oxygen include cancer and absolute contraindications of active viral disease, intercurrent pneumothorax and treatment with doxorubicin or cisplatin. This technique is particularly useful in the treatment of intractable hemorrhagic cystitis secondary to pelvic radiation therapy. Further investigation of the efficacy of hyperbaric oxygen is warranted for patients with necrotizing fasciitis (Fournier's gangrene), posttraumatic ischemic injury and/or impaired wound healing. CONCLUSIONS: Hyperbaric oxygen is a therapeutic alternative which complements the surgical and medical options for select patients. PMID- 10458335 TI - Intraoperative frozen section monitoring of nerve sparing radical retropubic prostatectomy. AB - PURPOSE: We describe the clinical and pathological outcomes of intraoperative frozen sections performed on the posterolateral prostate margins during nerve sparing radical prostatectomy. MATERIALS AND METHODS: We developed a technique of bilateral nerve sparing, inking the posterolateral prostate margins and obtaining frozen sections. When tumor was seen on frozen section, the fascia and neurovascular bundle were widely excised before completing the vesicourethral anastomosis. We reviewed 142 radical retropubic prostatectomies performed by a single surgeon between 1992 and 1997. Patients were divided into group 1--nerve sparing procedure using our technique (48 patients), 2--planned unilateral nerve sparing without frozen sections (46) and 3--planned bilateral nerve sparing without frozen sections (48). Potency was measured implicitly by physician assessment and explicitly with the UCLA Prostate Cancer Index. Group comparisons were made for positive margins, biochemical recurrence and potency. Mean followup was 24.5, 43.8 and 39.4 months for groups 1, 2 and 3, respectively. RESULTS: Of the 48 group 1 patients 9 (18%) had adenocarcinoma in the frozen section specimen, prompting wide excision of the bundles. None of these patients had biochemical recurrence during a mean followup of 20.5 months. Both bundles were spared in the remaining 39 patients (82%). There was no difference in survival or time to biochemical recurrence between groups 1 and 2. Potency was significantly different between groups 1 and 2 (36 versus 13%, p = 0.001), even after age adjustment (p = 0.05). In contrast, potency did not differ between groups 1 and 3 (38 versus 40%). Preoperative stage, grade and prostate specific antigen level were similar among the 3 groups. CONCLUSIONS: We found a significant difference in potency rates adjusted for age between patients with and without frozen sections. Our results indicate that this technique can enhance the ability of the surgeon to monitor the nerve sparing procedure without compromising cancer control. PMID- 10458336 TI - Clinical and genetic characterization of pheochromocytoma in von Hippel-Lindau families: comparison with sporadic pheochromocytoma gives insight into natural history of pheochromocytoma. AB - PURPOSE: Families with von Hippel-Lindau disease have variable risk of pheochromocytoma. Patients with von Hippel-Lindau disease and pheochromocytoma identified by screening can have no characteristic signs or symptoms. Families with von Hippel-Lindau disease were screened and followed to describe the natural history of von Hippel-Lindau pheochromocytoma, and to correlate these findings with von Hippel-Lindau germline mutation. MATERIALS AND METHODS: Between 1988 and 1997, 246 individuals with von Hippel-Lindau disease were identified (von Hippel Lindau group). Between August 1990 and June 1997, 26 consecutive patients with sporadic pheochromocytoma were evaluated (sporadic group). RESULTS: A total of 64 patients with von Hippel-Lindau disease had manifestations of pheochromocytoma, including 33 newly diagnosed during screening at the National Institutes of Health and 31 previously treated (93 adrenal and 13 extra-adrenal pheochromocytomas). Germline von Hippel-Lindau gene missense mutation was associated with extra-adrenal pheochromocytoma, younger age at presentation and the only patient with metastases. Of the 33 newly diagnosed patients with von Hippel-Lindau disease 4 had pheochromocytoma 2 times (37 pheochromocytomas) during followup. Of these pheochromocytomas 35% (13 of 37) were associated with no symptoms, normal blood pressure and normal catecholamine testing. Comparison of urinary catecholamines in the von Hippel-Lindau and sporadic groups demonstrated increased epinephrine, metanephrines and vanillylmandelic acid in the sporadic group. Analysis of urinary catecholamine excretion in the von Hippel Lindau and sporadic groups together demonstrated a correlation between tumor size, and urinary metanephrines, vanillylmandelic acid, norepinephrine, epinephrine and dopamine. In 12 patients without signs or symptoms of pheochromocytoma 17 newly diagnosed pheochromocytomas were followed for a median of 34.5 months without morbidity. Median tumor doubling time was 17 months. CONCLUSIONS: Von Hippel-Lindau gene missense mutation correlated with the risk of pheochromocytoma in patients with von Hippel-Lindau disease. These findings support a von Hippel-Lindau disease clinical classification, wherein some families are at high risk for manifestations of pheochromocytoma. Von Hippel Lindau disease pheochromocytomas identified by screening were smaller and less functional than sporadic pheochromocytomas. PMID- 10458337 TI - Laparoscopic renal and adrenal surgery in obese patients: comparison to open surgery. AB - PURPOSE: The efficacy and morbidity of laparoscopic renal and adrenal surgery in comparison to open surgery in obese patients are unknown. This retrospective study was performed to compare the outcome of laparoscopic versus open renal and adrenal surgery in the markedly and morbidly obese patient (body mass index 30 or greater). MATERIALS AND METHODS: The study group comprised all obese patients undergoing laparoscopic renal and adrenal surgery (laparoscopic group) from August 1997 to February 1998 at our institution. The majority of procedures were performed using a retroperitoneoscopic approach via the flank. These patients were compared with all obese patients undergoing open renal and adrenal surgery (open group) from 1994 to 1998. Open group patients with factors precluding laparoscopic surgery were excluded from the study (mass greater than 10 cm., renal vein and/or inferior vena caval thrombus and extension outside Gerota's fascia). RESULTS: There were 21 obese patients in each group and baseline parameters were comparable between groups. Median body mass index in the laparoscopic and open groups was 34 and 31, respectively. Median surgical time between the laparoscopic (210 minutes) and open (185) groups was comparable (p = 0.16). However, the laparoscopic group had decreased blood loss (100 versus 350 ml., p<0.001), quicker resumption of oral intake and ambulation (less than 1 versus 5 days, p<0.001), decreased narcotic analgesic requirements (12 versus 279 mg., p<0.001), shorter median hospital stay (less than 1 versus 5 days, p<0.001) and quicker convalescence (3 versus 9 weeks, p<0.001). There were 6 complications in 4 laparoscopic cases and 14 in 9 open surgery cases (p = 0.16). CONCLUSIONS: Markedly obese patients have an increased risk of complications from surgery, regardless of the approach. Our data suggest that laparoscopic renal and adrenal surgery is technically feasible in the markedly and morbidly obese patient, and compared with open surgery results in significantly decreased blood loss, quicker return of bowel function, less analgesic requirement, shorter convalescence and reduced hospital stay. PMID- 10458338 TI - Modified anatrophic nephrolithotomy for management of staghorn calculi: is renal function preserved? AB - PURPOSE: We report the results of modified anatrophic nephrolithotomy in select patients with complex staghorn calculi. MATERIALS AND METHODS: From 1987 to 1997 modified anatrophic nephrolithotomy, including 1 bilateral procedure, was performed in 15 patients at San Francisco General Hospital. Preoperative imaging included excretory urography, sonography and computerized tomography. Preoperative and postoperative quantitative renal function was assessed with 99mtechnetium dimercapto-succinic acid renal scintigraphy and serum creatinine measurements. RESULTS: Mean patient age was 42 years and 11 of the 15 patients were male. Bilateral nephrolithotomy was performed in 1 patient on separate occasions. Average surgical time was 3.7 hours with blood loss of 325 ml. Length of hospital stay averaged 4 days. Residual stones were present after 3 procedures, and 1 of these patients required a secondary procedure. There were no other short-term complications. Renal function was not significantly altered. CONCLUSIONS: Modified anatrophic nephrolithotomy for staghorn renal stones rendered most patients stone-free with concomitant preservation of renal function. Because of its safety, efficacy and simplicity we believe that continued use of this procedure is warranted in select patients with complex renal stone disease. PMID- 10458339 TI - Percutaneous stone removal in horseshoe kidneys. AB - PURPOSE: We review our experience with percutaneous nephrolithotomy for stones in horseshoe kidneys when extracorporeal shock wave lithotripsy was unsuccessful or inappropriate because of stone burden. MATERIALS AND METHODS: Stone burden, nephrostomy and percutaneous surgical techniques, and clinical outcome of 12 consecutive patients undergoing percutaneous nephrolithotomy for calculi in horseshoe kidneys between 1988 and 1997 were reviewed. RESULTS: Complete stone removal was achieved by percutaneous methods alone in 9 patients (75%). More than 1 nephrostomy tract was required in 5 patients and additional intraoperative nephrostomies were necessary in 4. Flexible nephroscopy was essential in achieving complete stone removal in 9 of 11 patients (82%). Second look procedures for stone removal were required in 3 patients. No sepsis occurred and no transfusions were needed. CONCLUSIONS: Percutaneous nephrolithotomy is a safe and effective method of stone removal in patients with calculi in horseshoe kidneys. Multiple nephrostomy tracts, the use of flexible nephroscopy and second look procedures may be necessary to achieve optimum stone removal. PMID- 10458340 TI - Live donor nephrectomy in patients with fibromuscular dysplasia of the renal arteries. AB - PURPOSE: We determine the suitability of patients with fibromuscular dysplasia of the renal arteries as renal donors. MATERIALS AND METHODS: We evaluated 37 patients with fibromuscular dysplasia who were potential renal donors. Renal artery lesions were graded I (mild) to IV (severe). Of the patients 19 underwent donor nephrectomy and 18 were rejected as donors. Reasons for rejection included severity of disease or availability of a better donor. Nephrectomy was performed on the side with fibromuscular dysplasia when disease was unilateral or on the side with more advanced disease when the lesions were bilateral. Patients were followed at 1 month, 1 year and then yearly. RESULTS: Of the 19 patients undergoing donor nephrectomy serum creatinine increased by a mean of 0.4 mg./dl. (range 0.1 to 1.1) on postoperative day 1. Effective renal plasma flow of the remaining kidney increased by 25% on postoperative day 5 and by 29% after 1 year. After a mean followup of 4.5 years no patient had hypertension, proteinuria or any significant change in serum creatinine compared to baseline values. Of the 18 patients not undergoing nephrectomy 11 were contacted at a mean followup of 4 years, and none had hypertension, proteinuria or abnormal serum creatinine. CONCLUSIONS: Patients with fibromuscular dysplasia may be acceptable renal donors. The decision to use a patient with fibromuscular dysplasia as a donor is based on patient age, the availability of other suitable donors, and the extent and severity of disease. PMID- 10458341 TI - The role of intraoperative heparin in cyclosporine treated cadaveric renal transplant recipients. AB - PURPOSE: We analyze the effect of intraoperative heparin in cadaveric renal transplantation. MATERIALS AND METHODS: We examined the records of 100 consecutive cadaveric transplant recipients who received 5,000 units of heparin at the time of vascular clamping (group 1), and compared the incidence of graft thrombosis and postoperative hemorrhagic complications to 100 consecutive cadaveric transplant recipients who did not receive heparin (group 2). The groups were similar in terms of donor age, sex, number of transplant recipients, sensitization, multiple vessels, delayed graft function and human leukocyte antigen mismatch. All patients received cyclosporine based triple therapy immunosuppression. All explanted grafts were examined histologically to rule out hyperacute rejection. RESULTS: There was no statistical difference between the 2 groups. The overall incidence of graft thrombosis was 5% (6% in group 1 and 4% in group 2). There was a greater need for blood transfusion in group 1, with 25 patients requiring transfusion postoperatively versus 14 in group 2. CONCLUSIONS: Intraoperative heparin did not reduce the incidence of graft thrombosis in this retrospective study. It did increase the postoperative blood transfusion requirements. As a result of this analysis, we have abandoned its use. PMID- 10458342 TI - Reexamining the value of hematuria testing in patients with acute flank pain. AB - PURPOSE: Hematuria testing is routinely performed in patients with acute flank pain to screen for ureterolithiasis and to help determine the need for excretory urography. Unenhanced helical computerized tomography (CT) has recently been shown to be superior to excretory urography in diagnosing ureteral obstruction and can evaluate many other causes of flank pain. Given the speed, accuracy and safety of CT the value of hematuria testing for acute flank pain should be reexamined. MATERIALS AND METHODS: We reviewed the medical records of 267 consecutive patients with acute flank pain referred for unenhanced helical CT. Microscopic and dipstick urinalysis data were obtained in 195 patients. Using helical CT as the gold standard, we calculated the sensitivity, specificity, predictive value and accuracy of hematuria for diagnosing ureterolithiasis. RESULTS: Of the patients with ureterolithiasis 33% had 5 or less, 19% had 1 or less and 11% had no red blood cells (RBCs) per high power field. Of the patients without ureterolithiasis 24% had greater than 5 and 51% had greater than 1 RBC per high power field. Of the patients with ureterolithiasis 14% had a negative dipstick test and 1 RBC or less per high power field. There were 25 patients without ureterolithiasis who had CT abnormalities unrelated to the urinary tract, of whom 8 had greater than 1 RBC per high power field. CONCLUSIONS: Absence of hematuria in the setting of acute flank pain cannot exclude a diagnosis of ureterolithiasis and should not obviate other diagnostic testing. Even when strongly positive on microscopy, hematuria has insufficient positive predictive value for diagnosing ureterolithiasis and may be misleading as other serious conditions resulting in acute flank pain may yield a positive test. PMID- 10458343 TI - Time to stone passage for observed ureteral calculi: a guide for patient education. AB - PURPOSE: We analyze the natural history of stone passage in patients with ureterolithiasis, and define factors predictive of spontaneous passage. MATERIALS AND METHODS: A total of 75 patients with ureteral calculi were prospectively followed for stone passage. Clinical data included patient gender and age, stone size and location, pain medication requirements and interval to stone passage. Of the 75 patients 13 (17%) required intervention and 62 (83%) were followed until spontaneous stone passage. Stones requiring intervention were not included in the time to passage analysis. RESULTS: Of the 75 patients 41 (55%) had ureteral stones 2 mm. or smaller with an average time to stone passage of 8.2 days and only 2 (4.8%) required intervention, 18 (24%) had stones between 2 and 4 mm. with an average time to stone passage of 12.2 days and 3 (17%) required intervention, and 16 had stones 4 mm. or greater with an average time to stone passage of 22.1 days and 8 required intervention. For 95% of stones to pass it took 31 days for those 2 mm, or less, 40 days for those 2 to 4 mm. and 39 days for those 4 to 6 mm. Multivariate analysis revealed that size, location and side were statistically related to stone passage interval (p = 0.012). Stones that were smaller, more distal and on the right side were more likely to pass spontaneously and required fewer interventions. CONCLUSIONS: Interval to stone passage is highly variable and dependent on stone size, location and side. Degree of pain, and patient gender and age had no bearing on the time to stone passage. Of ureteral stones 95% 2 to 4 mm. pass spontaneously but passage may take as long as 40 days. Intervention may be required in 50% of ureteral calculi greater than 5 mm. PMID- 10458344 TI - Laparoscopic versus open pyeloplasty: assessment of objective and subjective outcome. AB - PURPOSE: We determine the subjective and objective durability of laparoscopic versus open pyeloplasty. MATERIALS AND METHODS: From August 1993 to April 1997, 42 patients underwent laparoscopic pyeloplasty (laparoscopy group) with a minimum clinical followup of 12 months (mean 22). Subjective outcomes and objective findings were compared to those of 35 patients who underwent open pyeloplasty (open surgery group) from August 1986 to April 1997 with a minimum clinical followup of 12 months (mean 58). We assessed clinical outcome based on responses to a subjective analog pain and activity scale. In addition, radiographic outcome was assessed based on the results of the most recent radiographic study. RESULTS: Of the 42 laparoscopy group patients 90% (38) were pain-free (26, 62%) or had significant improvement in flank pain (12, 29%) after surgery. Two patients had only minor improvement and 2 had no improvement in pain. Surgery failed in only 1 patient with complete obstruction. A patent ureteropelvic junction was demonstrated in 98% (41 of 42 patients) of the laparoscopy group on the most recent radiographic study (mean radiographic followup 15 months). Of the 35 open surgery group patients 91% were pain-free (21, 60%) or significantly improved (11, 31%) after surgery. One patient had only minor improvement and 2 were worse. CONCLUSIONS: Pain relief, improved activity level and relief of obstruction outcomes are equivalent for laparoscopic and open pyeloplasty. PMID- 10458345 TI - Endourological treatment of ureteroenteric strictures: efficacy of Acucise endoureterotomy. AB - PURPOSE: The Acucise endoureterotomy balloon catheter has proved to be a safe and effective alternative to open surgery for the management of ureteropelvic junction obstruction and benign ureteral stricture disease. The established management of ureteroenteric strictures following urinary diversion is open surgical revision. There are few reports evaluating the efficacy of Acucise endoureterotomy in patients with ureteroenteric strictures. MATERIALS AND METHODS: The Acucise cutting balloon catheter was used to treat 9 patients with 12 ureteroenteric strictures 2 cm. or less in length. Median time from diversion to stricture was 48 months (range 13 to 192). Success was defined as resolution of symptoms and radiographic confirmation of patency. RESULTS: Recurrence of malignancy accounted for failure in 1 patient with bilateral strictures. Of the remaining 10 benign strictures 7 (70%) required open revision or serial stent changes. The remaining 3 strictures in 2 patients have remained patent at 30 and 18 months of followup for a success rate of 30%. There were no perioperative or operative complications. Mean followup was 9 months after Acucise treatment before intervention was necessary. CONCLUSIONS: Acucise endoureterotomy for ureteroenteric strictures following urinary diversion is a low morbidity procedure. Although the success rate is only 30%, Acucise endoureterotomy offers an alternative to immediate open surgical revision. PMID- 10458346 TI - An assessment of the use of intravesical potassium in the diagnosis of interstitial cystitis. AB - PURPOSE: We evaluated the use of intravesical potassium in the diagnosis of interstitial cystitis. MATERIALS AND METHODS: A blinded test assessment on 39 consecutive subjects attending our urology clinic for the evaluation of symptoms consistent with interstitial cystitis was performed. The pain response to intravesical potassium and water as a control was measured. The response rate was compared to the results of cystoscopy using standard outcome measures associated with diagnostic test assessment. RESULTS: The probability of having interstitial cystitis given a positive intravesical potassium test was 66%. This finding added no new useful information and would not be helpful with clinical decisions as the probability of having interstitial cystitis in this population was already 56% before the test. Similarly, if the test was negative then 46% or nearly half of the subjects were still likely to have interstitial cystitis. Therefore, a negative test would have no ability to rule out disease nor would it be useful in making clinical decisions about how to proceed with evaluation or therapy. Test characteristics were considered poor with a sensitivity of 69.5% and a specificity of 50%. Likelihood ratios (positive 1.39, negative 0.61) also indicated poor inclusion and exclusion capabilities. CONCLUSIONS: The general use of intravesical potassium as a diagnostic test for interstitial cystitis is not validated. The diagnosis of interstitial cystitis must depend on the clinical presentation and endoscopic findings based on National Institutes of Health criteria. PMID- 10458347 TI - Recurrence and progression in low grade papillary urothelial tumors. AB - PURPOSE: We report long-term followup data on patients with World Health Organization (WHO) grade I bladder tumors, and determine whether histopathological subgrouping as papillary neoplasm of low malignant potential and low grade papillary carcinoma is of clinical value. MATERIALS AND METHODS: All 680 patients in western Sweden with first diagnosis of bladder carcinoma in 1987 to 1988 were registered and followed for at least 5 years. Of the tumors 255 (37.5%) were stage Ta, WHO grade I. Tumors were further classified as papillary neoplasm of low malignant potential in 95 patients and low grade papillary carcinoma in 160 according to WHO and the International Society of Urological Pathology consensus classification of urothelial (transitional cell) neoplasms of the bladder. RESULTS: Mean age of patients at first diagnosis of low grade papillary carcinoma was 69.2 years, which was 4.6 years higher than those with papillary neoplasm of low malignant potential (p<0.005). During a mean observation time of 60 months our 255 patients underwent 577 operations for recurrences and had 1,858 negative cystoscopies. The risk of recurrence was significantly lower in patients with papillary neoplasm of low malignant potential compared to those with low grade papillary carcinoma (35 versus 71%, p<0.001). The risk of recurrence was higher in patients with multiple tumors at first diagnosis as well as those with recurrence at the first followup after 3 to 4 months. Stage progressed in 6 patients (2.4%), all with low grade papillary carcinoma at diagnosis. CONCLUSIONS: More than 90% of patients with stage Ta, WHO grade I have a benign form of bladder neoplasm, and few have truly malignant tumors. Future research should focus on reducing the number of recurrences and followup cystoscopies, and finding methods to identify malignant tumors so that pertinent treatment can be instituted. Subgrouping of WHO grade I bladder tumors as papillary neoplasm of low malignant potential and low grade papillary carcinoma seems to add valuable prognostic information. PMID- 10458348 TI - Is transurethral biopsy of the bladder necessary after 3 months to evaluate response to bacillus Calmette-Guerin therapy? AB - PURPOSE: We assessed the need for transurethral biopsy of the bladder at the 3 month evaluation in patients who have received intravesical bacillus Calmette Guerin (BCG). MATERIALS AND METHODS: A total of 81 patients collectively received 83, 6-week courses of BCG between 1995 and 1997, and underwent transurethral bladder biopsies 3 months after initiation of immunotherapy. The pretreatment and posttreatment bladder biopsy results, urinary cytology and posttreatment cystoscopic findings were correlated. RESULTS: Transitional cell carcinoma was demonstrated in 5 of 11 patients with erythematous bladder mucosa and a positive cytology compared to none of 37 with an erythematous lesion and a negative cytology. Of 12 patients with a papillary lesion on cystoscopy and a positive biopsy 10 had a negative cytology, and only 1 of 13 patients with a negative cystoscopy had a positive biopsy. CONCLUSIONS: Bladder biopsy is not necessary in patients 3 months after receiving BCG who have a normal office cystoscopy or an erythematous bladder and normal urine cytology. PMID- 10458349 TI - A stage specific approach to tumor surveillance after radical cystectomy for transitional cell carcinoma of the bladder. AB - PURPOSE: Surveillance protocols after radical surgery for genitourinary tumors typically do not consider that the risk of recurrence is stage dependent. We describe the development of a stage specific protocol for monitoring patients with transitional cell carcinoma for tumor recurrence and conduit complications after radical cystectomy. MATERIALS AND METHODS: The records of 382 patients with transitional cell carcinoma who underwent cystectomy in 1986 to 1994 were reviewed for the dates and presenting symptoms of local and distant recurrences, and the results of radiological imaging studies and liver function tests. Based on the division of patients into pathological stages of pT1, pT2 and pT3 groups, we developed a new transitional cell carcinoma surveillance protocol. RESULTS: Of 97 patients with transitional cell carcinoma metastases 72 (74%) were asymptomatic, including 43 with metastases detected by routine chest x-rays (30) or blood tests (13). Surveillance computerized tomography identified isolated asymptomatic intra-abdominal metastases in 10 patients (10%), of whom 90% had pT3 disease. Based on these results we recommend a stage specific surveillance protocol for pT1--annual history, physical examination, chest x-ray and laboratory studies, pT2-same studies at 6, 12, 18, 24, 30, 36, 48 and 60 months after cystectomy, and pT3-same studies at 3, 6, 12, 18, 24, 30, 36, 48 and 60 months plus computerized tomography at 6, 12 and 24 months after cystectomy. A radiographic study of the upper tract should be performed in all patients every 1 to 2 years to evaluate for recurrences and complications of the ileoureteral anastomosis. CONCLUSIONS: A stage driven surveillance strategy for monitoring patients after radical cystectomy can reduce costly imaging studies while efficiently detecting recurrences and complications. PMID- 10458350 TI - Comparison of mechanical reliability of original and enhanced Mentor Alpha I penile prosthesis. AB - PURPOSE: We report the results of a prospective study of 1,381 Mentor Alpha I penile prostheses implanted to treat impotence, and compare original and enhanced penile prosthesis mechanical reliability. MATERIALS AND METHODS: The study consisted of 410 original models manufactured before November 1992 and 971 enhanced models manufactured since December 1992. Implants were further stratified as first time (virgin) or replacements of a previous implant. Mechanical failure-free survival rates for the original prosthetic and enhanced models were compared. RESULTS: The 5-year survival rate increased from 75.3% for the original to 92.6% for the enhanced model overall (log rank p<0.0001), and from 75.3 to 93.6%, respectively, for the virgin implants only (log rank p<0.0001). The estimated failure rate of approximately 5.6% for the original model was fairly consistent during followup, while the significantly lower failure rate of 1.3% for the enhanced model was not. The failure rate of the enhanced model implants was about 0.8% per year during the first 3.5 years and increased to approximately 3.1% per year thereafter. CONCLUSIONS: Our results strongly support the premise that mechanical reliability is superior with the enhanced compared to the original model. PMID- 10458351 TI - An assessment of the clinical relevance of serum testosterone level determination in the evaluation of men with low sexual drive. AB - PURPOSE: We evaluated the relationship between sexual drive and serum testosterone level among men with erectile dysfunction. MATERIALS AND METHODS: A retrospective medical record review of 180 men who attended a hospital based erectile dysfunction clinic between April 1997 and January 1998 was performed. Sexual drive was assessed using the Brief Sexual Function Inventory questionnaire, and serum total and free testosterone levels were measured. RESULTS: A total of 108 complete records were included in this report. Mean patient age plus or minus standard deviation was 59.5+/-10.2 years (range 33 to 79). Of the men 55 (50.9%) had low, 38 (35.2%) moderate and 15 (13.9%) high sexual drive. Mean sexual drive, bothersomeness and total sexual function scores among the groups were significantly different (p<0.001). Mean serum total testosterone levels among men with low, moderate and high sexual drive were 2.8, 3.2 and 3.4 mg./ml. (normal 2.8 to 8.8), respectively, and mean free testosterone levels were 9.1, 9.5 and 11.4 pg./ml. (normal 8.7 to 54.7), respectively. Differences among means were not statistically significant. As a screening test for low sexual drive, low total testosterone had a positive predictive value, negative predictive value and yield of 59.2, 55.9 and 26.9%, respectively. PMID- 10458352 TI - Clinical efficacy of sildenafil citrate based on etiology and response to prior treatment. AB - PURPOSE: We assess the clinical efficacy of sildenafil citrate and predictors of satisfactory outcome. MATERIALS AND METHODS: All patients treated with sildenafil citrate within the first 6 weeks of its release were evaluated with a self administered questionnaire before and at completion of therapy to assess etiology of erectile dysfunction, level of sexual function, libido, response to previous therapies, response to therapy with sildenafil citrate and quality of life. Sexual function was measured before and during therapy using an abbreviated version of the International Index of Erectile Function, with a successful outcome defined as a level of satisfaction of 4 or 5 on a 5-point scale. RESULTS: Followup was obtained in 267 of the 308 patients who entered the study. Mean age plus or minus standard deviation was 61+/-9.6 years and duration of erectile dysfunction was 4.1+/-3 years. Overall satisfaction with sildenafil citrate for the entire patient population was 65% and response to prior therapies did not affect satisfaction. There was a significant positive correlation between baseline sexual function and response to sildenafil citrate but even patients with severe erectile dysfunction had a 41% satisfaction rate. Etiology of erectile dysfunction had a significant impact on satisfaction rate, with neurogenic causes of erectile dysfunction (diabetes, prostate surgery and so forth) having significantly lower rates than psychogenic or vasculogenic erectile dysfunction. CONCLUSIONS: Sildenafil citrate is a highly effective oral agent for the treatment of erectile dysfunction in clinical practice. The best predictors for response to sildenafil citrate therapy are baseline sexual function and etiology of erectile dysfunction. However, we could not identify any patient characteristic that would predict absolute failure for sildenafil citrate therapy. Therefore, all patients with erectile dysfunction who do not have specific contraindications should be considered for sildenafil citrate therapy. PMID- 10458353 TI - Topical prostaglandin E1 SEPA gel for the treatment of erectile dysfunction. AB - PURPOSE: We compare the systemic effects, local tolerance and effectiveness of topical gel formulations on the penis containing alprostadil (prostaglandin E1) plus 5% SEPA versus SEPA alone (placebo) in men with erectile dysfunction. MATERIALS AND METHODS: Erectile response, skin discomfort and erythema were measured in 48 men with erectile dysfunction secondary to vascular, neurogenic, psychogenic or mixed etiologies in this single-blind, placebo controlled trial. RESULTS: Application of prostaglandin E1 gel correlated positively with erectile response as 67 to 75% of patients had an erection compared to 17% of controls (p<0.001). Blood pressure and heart rate varied minimally. No serious adverse effects were observed in the 48 patients, although the majority had skin discomfort. CONCLUSIONS: Topical prostaglandin E1 gel applied to the penis appears to be safe, and facilitates audiovisual and tactile stimulation resulting in an erection when given in a clinic setting. Consequences to the female partner remain unknown. PMID- 10458354 TI - Medical therapies for erectile dysfunction. PMID- 10458355 TI - Laparoscopic testicular denervation for chronic orchalgia. AB - PURPOSE: No specific cause is identified in most cases of chronic orchialgia. Nonsurgical therapies, including management at a chronic pain clinic, are generally recommended. Only when multiple conservative measures fail are patients offered surgical intervention, such as orchiectomy. We evaluate laparoscopic testicular denervation as an organ preserving and minimally invasive surgical alternative. MATERIALS AND METHODS: Since 1993, 9 patients with chronic orchialgia have undergone transperitoneal laparoscopic testicular denervation after nonsurgical modalities failed. Using 1, 10 mm. and 1 or 2, 5 mm. ports, the gonadal vessels were isolated circumferentially and divided cephalad to the vas deferens and its vasculature. Preoperative treatment modalities, morbidity and outcome were documented. A cord block provided temporary relief in all 9 patients. Analog scales were used to assess long-term pain relief (0 no pain to 100 worst pain) and activity levels (0 bedrest to 100 no restrictions). RESULTS: Average symptom duration before laparoscopic testicular denervation was 4.1 years. Of 9 patients 8 had undergone prior scrotal surgery. Failed nonsurgical modalities included anti-inflammatory drugs in 7 patients, antibiotics in 6, pain clinic consultations in 4 and antidepressant medications in 2. Mean pain score decreased from 69.4 (range 35 to 90) preoperatively to 30.6 at a mean followup of 25.1 months. Excluding the 2 cases that had no pain relief (less than 10-point reduction), average pain score decreased from 69 to 19 postoperatively (mean reduction 71%). Activity levels improved in all cases. There were no significant complications, including testis atrophy. One patient who had no pain relief underwent subsequent hydrocelectomy for pain, which also failed. CONCLUSIONS: Laparoscopic testicular denervation can provide significant long-term pain relief and appears to be a reasonable alternative in select cases with chronic orchialgia refractory to medical therapy. Larger series and prospective evaluations are necessary. PMID- 10458356 TI - Varicocele repair improves semen parameters in azoospermic men with spermatogenic failure. AB - PURPOSE: We analyzed the efficacy of varicocele repair in improving semen parameters in azoospermic men with spermatogenic failure. MATERIALS AND METHODS: After routine clinical evaluation with confirmatory pellet analysis testicular biopsy and varicocele repair were simultaneously performed in 28 azoospermic men with a primary diagnosis of unilateral or bilateral varicocele and spermatogenic failure. Semen analyses were obtained starting 4 months after varicocelectomy. RESULTS: Repair was bilateral repair in 20 men and unilateral in 8. Of the 28 men 12 (43%) had sperm in the ejaculate with a mean postoperative sperm count plus or minus standard deviation of 1.2+/-3.6x10(6)/ml. and an average followup of 24 months. Mean sperm motility was 19+/-24% (range 0 to 80). Testicular biopsy was predictive of outcome. Only 9 men with severe hypospermatogenesis and 5 with maturation arrest spermatid stage had improvement in sperm density. No improvement was seen in 3 men with the Sertoli-cell-only pattern or 3 with maturation arrest spermatocyte stage. No pregnancies by natural intercourse resulted. One couple used fresh ejaculate for intracytoplasmic sperm injection and 1 underwent testicular sperm extraction with intracytoplasmic sperm injection. Both pregnancies resulted in live births. No other predictive factors were identified. CONCLUSIONS: Varicocele repair can result in sperm in the ejaculate of azoospermic men when severe hypospermatogenesis or maturation arrest spermatid stage is present. Select men with spermatogenic failure and varicoceles may be candidates for varicocele repair, rather than resorting to testis biopsy for sperm extraction in preparation for intracytoplasmic sperm injection. However, the couple should be counseled that assisted reproductive technologies will most likely be required to initiate pregnancy. PMID- 10458357 TI - Serial prostate specific antigen screening for prostate cancer: a computer model evaluates competing strategies. AB - PURPOSE: We compare prostate specific antigen (PSA) screening strategies in terms of expected years of life saved with screening, number of screens, number of false-positive screens and rates of over diagnosis, defined as detection by PSA screening of patients who would never have been diagnosed without screening. MATERIALS AND METHODS: A computer model of disease progression, clinical diagnosis, PSA growth and PSA screening was used. Under baseline conditions, when screening is not considered, the model replicates clinical diagnosis and disease mortality rates recorded by the Surveillance, Epidemiology and End Results Program of the National Cancer Institute in the mid 1980s. RESULTS: Biannual screening with PSA greater than 4.0 ng./ml. was projected to reduce the number of screens and false-positive tests by almost 50% relative to annual screening while retaining 93% of years of life saved. With annual screening use of an age specific bound for PSA to consider a test positive instead of the standard 4.0 ng./ml. was projected to reduce false-positive screens by 27% and over diagnosis by a third while retaining almost 95% of years of life saved. Sensitivity analyses did not change the relative efficacy of biannual screening. CONCLUSIONS: Under the model assumptions biannual PSA screening is a cost-effective alternative to annual PSA screening for prostate cancer. With annual screening use of an age specific bound for PSA positivity appears to reduce false-positive results and over diagnosis rates sharply relative to a bound of 4 ng./ml. while retaining most of the survival benefits. PMID- 10458358 TI - Influence of sextant prostate needle biopsy or surgery on the detection and harvest of intact circulating prostate cancer cells. AB - PURPOSE: The feasibility of harvesting intact, circulating prostate cancer cells from the blood of men with advanced prostate cancer has previously been demonstrated. We studied the influence of sextant prostate needle biopsy and radical prostatectomy on harvesting intact circulating prostate cancer cells. MATERIALS AND METHODS: Via standard venipuncture 20 c.c. blood were obtained preoperatively, and 30 minutes and 3 days postoperatively from 23 men with clinically localized prostate cancer undergoing surgery. Similarly, blood was obtained before and after routine prostate biopsy from 13 men for an elevated prostate specific antigen level and/or abnormal digital rectal examination. The blood cells were removed via density centrifugation and magnetic cell sorting. The remaining prostate epithelial cells were characterized by indirect fluorescent immunocytochemical staining and fluorescent in situ hybridization using deoxyribonucleic acid probes. RESULTS: Sextant biopsy of the prostate induced circulating cells in 3 of 13 men (23%), only 1 of whom demonstrated cells with aneuploidy (Gleason score 3+4 = 7). Circulating cells were detected preoperatively, 30 minutes or 3 days postoperatively in 35% of radical prostatectomy cases. Of the patients 13% had detectable circulating cells 30 minutes postoperatively only and 9% had cells harvested on postoperative day 3. Persistence of circulating prostate cancer cells was noted in 13% of men on postoperative day 3. Serum prostate specific antigen level and pathological stage did not appear to be related to harvested cell number. CONCLUSIONS: Prostate cancer cells can be harvested from men with clinically localized disease undergoing sextant needle biopsy or radical prostatectomy. Routine prostate biopsy and surgery may influence the number of measurable circulating cells in the short term but the clinical significance and long-term prevalence of detectable circulating cells are unknown. Further studies are needed to evaluate the clinical usefulness of this assay for detecting, staging and monitoring prostate cancer. PMID- 10458359 TI - Effect of neoadjuvant hormonal therapy on prostatic intraepithelial neoplasia and its prognostic significance. AB - PURPOSE: We studied the effect of neoadjuvant hormonal therapy on prostatic intraepithelial neoplasia in patients undergoing radical prostatectomy and assessed the effect of prostatic intraepithelial neoplasia on disease recurrence as measured by serum prostate specific antigen (PSA). MATERIALS AND METHODS: A total of 278 patients with clinically localized prostate cancer were included in phase II and III studies evaluating radical prostatectomy alone versus radical prostatectomy following neoadjuvant hormonal therapy at Memorial Sloan-Kettering Cancer Center between October 1991 and August 1996. Patient data related to prostatic intraepithelial neoplasia were analyzed. RESULTS: Of 275 evaluable patients 145 (52.7%) had prostatic intraepithelial neoplasia. Of 50 patients treated with neoadjuvant hormonal therapy (hormone group) 22 (44%) had a lower incidence of prostatic intraepithelial neoplasia compared to 69 of 80 controls (86.3%) (chi-square test p<0.0001). Of 262 patients (95.3%) with followup PSA 44 (16.8%) had PSA recurrence at a median followup of 32 months, with a median time to recurrence of 30 months. PSA recurrence was noted in 23 of 145 patients with compared to 21 of 130 without prostatic intraepithelial neoplasia (chi-square test p = 0.95), and did not significantly differ between the hormone group (25 of 142, 17.6%) and controls (19 of 130, 14.6%) (chi-square test p = 0.45). CONCLUSIONS: While patients treated with neoadjuvant hormonal therapy had significantly lower incidence of prostatic intraepithelial neoplasia, neither prostatic intraepithelial neoplasia nor neoadjuvant hormonal therapy significantly affected PSA recurrence at a median followup of 32 months. PMID- 10458360 TI - The durability of external beam radiation therapy for prostate cancer: can it be identified? AB - PURPOSE: We establish criteria to identify a durable response to external beam radiation therapy by calculation of biochemical progression-free probability for patients who attained and maintained defined nadir prostate specific antigen (PSA) levels more than 5 years after treatment. MATERIALS AND METHODS: A total of 460 patients were treated with external beam radiation monotherapy from 1976 to 1995. Patients with PSA less than 0.5 (group 1) or 0.5 to 1.0 (group 2) ng./ml. more than 5 years after treatment were identified. Treatment failure was defined as 3 consecutive increases in PSA after nadir. Progression-free probability after 60 months was calculated for each group. A comparison was also made to patients achieving the same nadir levels anytime after treatment. RESULTS: Failure occurred at 133 months in 1 of 26 group 1 patients (4%) and at a median of 76 months in 5 of 26 group 2 patients (19%). At 10 years progression-free probability was 91% for group 1 compared to 72% for group 2 (p = 0.0575). These same nadir levels anytime after treatment were associated with higher failure rates of 55% for group 1 and 72% for group 2. CONCLUSIONS: If a PSA nadir of less than 0.5 ng./ml. was maintained 5 years after therapy, subsequent failure was rare. Although statistical significance was not reached (p = 0.0575), a higher failure rate was noted if the nadir PSA was 0.5 to 1.0 ng./ml. at 5 years. Thus, patients with PSA 0.5 to 1.0 ng./ml. require careful continued surveillance. Nadir levels less than 1.0 ng./ml. anytime before 5 years were associated with a substantial risk of subsequent progression. PMID- 10458361 TI - Is there a role for digital rectal examination in the followup of patients after radical prostatectomy? AB - PURPOSE: We determine the role of digital rectal examination in the followup of patients after radical prostatectomy. MATERIALS AND METHODS: We retrospectively analyzed data on 501 consecutive patients who underwent radical retropubic prostatectomy between 1992 and 1998, and were followed at the University of Miami. Patients were evaluated at 3 to 6-month intervals after surgery with serum prostate specific antigen (PSA) and digital rectal examination. Biochemical recurrence was defined as PSA greater than 0.2 ng./ml. and increasing on at least 2 consecutive measurements. Local recurrence, detected by an abnormal digital rectal examination, was defined as an induration or nodularity in the prostatic fossa. RESULTS: Mean followup plus or minus standard deviation was 25.4+/-20.8 months. Disease recurred in 72 patients (14.4%) and was biochemical in all. An abnormal digital rectal examination was noted in 4 patients, none of whom had an undetectable PSA at the time of a palpable abnormality. CONCLUSIONS: Our results suggest that an abnormal digital rectal examination after radical prostatectomy is always associated with a detectable PSA, which implies that performing a digital rectal examination in the absence of a detectable PSA may not be necessary. PMID- 10458362 TI - Complications of laparoscopic procedures in urology: experience with 2,407 procedures at 4 German centers. AB - PURPOSE: The 4 most active centers of the laparoscopy working group of the German Urologic Association collected data about the complications associated with laparoscopic surgery in urology. MATERIALS AND METHODS: At 4 centers 2,407 laparoscopies or retroperitoneoscopies were performed as of May 1998, including 776 for varicocelectomy, 259 for cryptorchidism, 481 for pelvic lymph node dissection, 351 for nephrectomy/heminephrectomy renal pathology, 139 for renal cyst resection, 58 for ureteral procedures, 44 for adrenalectomy, 41 for nephropexy, 41 for lymphocele fenestration, 40 for retroperitoneal para-aortic lymphadenectomy and 187 for other operations. The complications were evaluated, listed according to the anatomical specificity and grouped with respect to the surgical step during laparoscopy. RESULTS: A total of 107 complications (4.4%) occurred. The re-intervention rate was 0.8% and the mortality rate was 0.08%. The complication rate depended on the difficulty of the procedure and averaged 1.0, 3.9 and 9.2%, respectively, for easy, difficult and very difficult operations. The majority were vascular injuries (1.7%) and visceral lesions (1.1%) followed by complications of healing and infection (0.8%). Only 0.2% of complications was associated with the access technique (trocar insertion), whereas most occurred during dissection (2.9%). The complication rate was 13.3% for the first 100 procedures and subsequently averaged 3.6%. CONCLUSIONS: Critical documentation of experience from several institutions, especially for an analysis of complications of urological laparoscopy, is important for the development of this surgical technique. The overall complication rate is comparable to other specialties. Future technical developments in trocar insertion, tissue dissection and control of bleeding with our improved training program will further reduce the complication rate. PMID- 10458363 TI - Reconstructive renal surgery using a water jet. AB - PURPOSE: We describe the successful application of a water jet to reconstructive renal surgery. MATERIALS AND METHODS: Two consecutive patients underwent reconstructive renal surgery using a waterjet apparatus for a 4.0x6.5 cm. well encapsulated tumor of the lower pole of the left kidney and a 4.5x2.5 cm. staghorn calculus of the left kidney, respectively. The water jet apparatus (Parenchimotom 01) consists of a pressure generating pump and a flexible hose connected to the hand piece, and a nozzle with a pinhole opening of 0.3 mm. RESULTS: Both patients underwent surgery through a left lumbar incision. Partial nephrectomy was performed in 1 patient and anatrophic nephrotomy for stone removal in the other. Dissection time was 25 and 12 minutes, with blood loss of 150 and 100 ml., respectively. No temporary vascular clamping or local hypothermia was necessary. Both patients were discharged home 10 days after surgery and at followup no negative sequelae were attributable to the procedure. CONCLUSIONS: The operative procedures proved easy, fast and effective. No temporary vessel clamping or hypothermia was required. Using a water jet is a novel approach that is helpful in renal surgery. PMID- 10458364 TI - Stone entrapment during percutaneous removal of infection stones from a continent diversion. PMID- 10458365 TI - Percutaneous vesicolithotomy: an alternative to open bladder surgery in patients with an impassable or surgically ablated urethra. AB - PURPOSE: Although vesical calculi are routinely treated transurethrally, open vesicolithotomy is generally performed in patients with an impassable or surgically ablated urethra. We describe a technique of percutaneous vesicolithotomy which we used in patients who had undergone urethral ablation and concomitant continent diversion by appendicovesicostomy. MATERIALS AND METHODS: Bladder stones were detected in 3 patients with neurogenic bladder who had undergone continent urinary diversion with bladder neck closure and appendicovesicostomy. To treat the stones access to the bladder was achieved percutaneously and the tract was enlarged using a balloon dilator. An Amplatz sheath was slipped over the inflated balloon and after the dilator was removed the sheath provided a working channel through which stones were fragmented and removed using a nephroscope. RESULTS: Each patient was rendered stone-free and discharged home the same day as the procedure. CONCLUSIONS: Percutaneous vesicolithotomy provides an alternative approach for bladder stone removal in patients with an impassable urethra with decreased morbidity compared to open procedures. PMID- 10458366 TI - Collagen injections for urinary stress incontinence in a small urban urology practice: time to failure analysis of 99 cases. AB - PURPOSE: We examined the durability of collagen injection response for female incontinence using Kaplan-Meier survival statistics and the log rank test. MATERIALS AND METHODS: A total of 99 women 26 to 84 years old (mean age 60) underwent collagen injections for urethral incompetence. In this retrospective chart audit treatment failures were analyzed using survival statistics. In addition, we conducted a sensitivity analysis of the data using a worst and best case model to account for variability of the data due to the high amount of censored observations. RESULTS: Mean incidence of success was 56% with a mean followup of 9.5 months (95% confidence interval 0.17 to 24.6). Median duration of success was 4.7 months (95% confidence interval 3.4 to 5.9). Of the 10 time to failure function comparisons only cystometric incontinence classification had a statistically significant predictive value for treatment failures (p = 0.003). The survival analysis curve provided a 13% success probability at 18 months and was close to the worst case time to failure function. CONCLUSIONS: Collagen injection has minimal morbidity and a low median success rate. Bladder instability, as determined by cystometrogram, is an important determinant of success. Survival statistics methods should be used more widely to describe urological outcomes because they provide dynamic and, thus, more meaningful information to urologists and patients than summary statistics. PMID- 10458367 TI - Intravascular capillary hemangioma presenting as a solid renal mass. PMID- 10458368 TI - Renal artery thrombosis secondary to ureteral obstruction. PMID- 10458369 TI - Indwelling ureteral stent fragmentation with severe encrustation and stone formation. PMID- 10458370 TI - Retroperitoneal laparoscopic marsupialization of periureteral urinoma. PMID- 10458371 TI - The giant of the giant schwannomas. PMID- 10458372 TI - The largest bladder diverticulum ever? PMID- 10458373 TI - Xanthogranulomatous cystitis associated with anaerobic bacterial infection. PMID- 10458374 TI - Urothelial carcinoma in a suprapubic cystostomy tract 27 years after tube removal. PMID- 10458375 TI - Hepatocellular carcinoma metastatic to the bladder after liver transplantation. PMID- 10458376 TI - Giant hepatoid yolk sac tumor in a black man. PMID- 10458377 TI - Phyllodes type of atypical prostatic hyperplasia. PMID- 10458378 TI - Estramustine phosphate withdrawal syndrome with dramatic pain relief. PMID- 10458379 TI - Re: A randomized controlled trial to assess the incidence of new onset hypertension in patients after shock wave lithotripsy for asymptomatic renal calculi. PMID- 10458380 TI - Re: Hand assisted laparoscopic nephrectomy: comparison to standard laparoscopic nephrectomy. PMID- 10458381 TI - Re: The diagnosis of interstitial cystitis revisited: lessons learned from the National Institutes of Health Interstitial Cystitis Database Study. PMID- 10458382 TI - Re: Cystoscopic findings consistent with interstitial cystitis in normal women undergoing tubal ligation. PMID- 10458383 TI - Re: Hydronephrosis as a prognostic indicator in bladder cancer patients. PMID- 10458384 TI - Re: Recurrent bilateral amiodarone induced epididymitis. PMID- 10458385 TI - Re: Metastatic adenocarcinoma of the prostate to the choroid with loss of visual acuity as a presenting symptom. PMID- 10458386 TI - Natural history of fetal simple renal cysts detected in early pregnancy. AB - PURPOSE: In this 12-year prospective, longitudinal study we investigated the natural history of fetal simple renal cysts identified by ultrasonography in early pregnancy. MATERIALS AND METHOD: A detailed sonographic examination of the fetus was performed between January 1987 and June 1998 in 29,984 consecutive pregnancies at 14 to 16 weeks of gestation. Amniocenteses and chromosomal investigations were done in all cases in which a simple renal cyst was detected in the fetus. Followup sonography was done in all cases of renal cyst during pregnancy, infancy and, when indicated, childhood. RESULTS: Simple renal cysts were diagnosed at 14 to 16 weeks of gestation in 28 fetuses (1/1,100 pregnancies, 0.09%). In 25 fetuses the cysts resolved during pregnancy. In 2 fetuses the cysts remained benign but persisted postnatally and in 1 a renal cyst that was initially defined as simple was the first sign of unilateral multicystic dysplastic kidney. Except for nonseptated cystic hygroma in 1 fetus, none of the others had associated anomalies of the urinary or other organ systems and no chromosomal anomalies. Postnatal followup in all cases revealed healthy children. CONCLUSIONS: A fetal simple renal cyst can be identified by ultrasonography in early pregnancy. In the absence of associated anatomical or chromosomal abnormalities, the majority of cysts will resolve during pregnancy without any sequelae. Given the transient nature of most fetal simple renal cysts detected in early pregnancy, it is possible that these cysts represent a distinct entity within the spectrum of cystic kidney diseases. PMID- 10458387 TI - Adjunctive oral corticosteroids reduce renal scarring: the piglet model of reflux and acute experimental pyelonephritis. AB - PURPOSE: We investigate the efficacy of antibiotics combined with corticosteroid in diminishing post-pyelonephritic renal scarring compared to standard antibiotic therapy. MATERIALS AND METHODS: Bilateral vesicoureteral reflux was surgically created in 36 piglets (72 kidneys). A week later each bladder was inoculated by percutaneous injection with a standardized broth culture of Escherichia coli and molten paraffin. 99mTechnetium dimercapto-succinic acid (DMSA) scintigraphy was performed 3 days after introduction of urinary infection to detect the presence of acute pyelonephritis. Acute pyelonephritic lesions seen on DMSA scans were graded according to the percentage of renal zone involvement as grade 1--less than 33%, grade 2--33 to 66% and grade 3--greater than 66% involved. When pyelonephritis was present, piglets were randomized to receive either standard antibiotics or antibiotics and 2 mg./kg. prednisolone daily. 99mTechnetium-DMSA scintigraphy was repeated 2 months after completion of therapy, and the kidneys were harvested for gross and histopathological examination. Each kidney was divided into upper, middle and lower zones for correlation of pathological and imaging findings. Severity of renal scarring was then assessed using histopathological confirmation of gross anatomical findings as grade 1--less than 1, grade 2-1 to 2 and grade 3-greater than 2 cm. RESULTS: Acute pyelonephritis was induced in 136 of 216 renal zones. The sites of renal scarring corresponded anatomically to sites of acute pyelonephritis in all but 5 cases. Overall, the prevalence of post-pyelonephritic scarring was 56.6% (77 of 136) of renal zones. The severity of scarring in both groups correlated with the severity of the initial pyelonephritic lesion. Of the 31 zones that formed grade 3 renal scars the distribution of grades 1, 2 and 3 acute pyelonephritis on the initial DMSA scan was 3, 26 and 71%, respectively. Grade 3 acute pyelonephritis was more likely to result in severe (grade 3) renal scars in the control compared to the steroid treated group (59 versus 31%). Overall, acute pyelonephritis completely resolved in 40% of controls and 51% of steroid treated animals. However, only 9% of control animals with grade 3 acute pyelonephritis demonstrated complete resolution, as opposed to 28% of those receiving steroids. CONCLUSIONS: The risk of renal scarring is greatest after severe acute pyelonephritis involving greater than 66% of a renal zone. Adjunctive oral prednisolone appears to be effective in diminishing renal scarring in severely affected kidneys. In kidneys with mild and moderate acute pyelonephritis antibiotics alone appear to be equally effective in preventing scarring. PMID- 10458388 TI - The role of 99mtechnetium dimercapto-succinic acid renal scans in the evaluation of occult ectopic ureters in girls with paradoxical incontinence. AB - PURPOSE: We describe the use of 99mtechnetium (Tc) dimercapto-succinic acid (DMSA) renal scintigraphy to document poorly functioning and/or ectopic renal units associated with occult ectopic ureters. MATERIALS AND METHODS: During the last 8 years 6 toilet-trained girls were referred for lifelong continuous urinary dribbling. Initial radiological evaluation included renal and bladder sonography in 6, excretory urography in 4, a mercaptoacetyltriglycine-3 renal scan in 1 and cystovaginoscopy in 2. Ultimately a 99mTc-DMSA renal scan was performed in all 6 girls before surgical intervention. RESULTS: Sonography failed to establish the diagnosis of ureteral ectopia in all cases, and revealed a normal solitary kidney in 3, normal kidneys in 1, an apparently uncomplicated unilateral duplication without hydroureteronephrosis in 1 and a contralateral uncomplicated duplication in 1. Excretory urography in 4 cases was inconclusive and showed a solitary kidney in 1, ipsilateral duplication without a normal appearing upper pole collecting system in 1, contralateral uncomplicated duplication in 1 and normal kidneys in 1. A mercaptoacetyltriglycine-3 renal scan in another girl with a solitary kidney on sonography failed to demonstrate a contralateral small ectopic kidney. Cystovaginoscopy performed in 2 patients by other pediatric urologists was nondiagnostic. 99mTc-DMSA renal scintigraphy was diagnostic in all 6 cases, and revealed a small poorly functioning ectopic kidney in 3 and a poorly functioning dysplastic upper pole moiety in 3, which were consistent with a diagnosis of ureteral ectopia. An ectopic ureter was confirmed by cystoscopic and surgical findings in all girls. CONCLUSIONS: 99mTc-DMSA renal scintigraphy reliably detects and localizes hypoplastic ectopic kidneys and poorly functioning upper pole moieties associated with occult ectopic ureters in girls with continuous urinary leakage. PMID- 10458389 TI - Salvage continent vesicostomy after enterocystoplasty in the absence of the appendix. AB - PURPOSE: We describe a surgical procedure for a select group of children who had previously undergone augmentation enterocystoplasty, following which intermittent catheterization became more and more difficult or impractical. A new access to the reservoir became necessary, and alternative conduits included the appendix, ureter and tubularized ileum or stomach. Each conduit had its advantages and disadvantages but all required transperitoneal dissection. We report a simple extraperitoneal surgical technique that involves use of the Mitrofanoff and Nissen principles. MATERIALS AND METHODS: A 2x6 cm. flap of the anterior wall, which is usually intestine, of the augmented bladder is raised. The base of the flap is just below the bladder dome. The flap is then tubularized over a 14F catheter and the cystostomy is closed. The bladder is plicated around the base of the tube, similar to the Nissen gastroesophageal fundoplication. The plication extends and covers the proximal 3 cm. of the tube. Intraoperative bladder distention is performed to confirm the competence of the continence mechanism. The distal part of the tube is then anastomosed to the inverted umbilical skin or to a tubularized abdominal wall skin flap. RESULTS: The aforementioned technique was used in 5 children 3 months to 6 years old. Earlier augmentation ileocystoplasty and bladder neck reconstruction had been performed in 4 children. The appendix was unavailable for a Mitrofanoff vesicostomy in all cases. This procedure was also performed on a 13-year-old boy with severe myogenic detrusor failure, due to posterior urethral valves, and a bladder capacity of 700 ml. There was no significant surgical morbidity and all children are dry between clean intermittent catheterizations 1 to 5 years postoperatively (mean 3.3). CONCLUSIONS: Use of the Mitrofanoff and Nissen principles proved to be simple and reliable, and avoided secondary intraperitoneal exploration and use of bowel in these select cases. PMID- 10458390 TI - Progressive dilation for bladder tissue expansion. AB - PURPOSE: The use of gastrointestinal tissue for augmentation cystoplasty is associated with numerous complications. We previously reported the development of a system in which ureters were progressively dilated and used for ureterocystoplasty. We have now applied a similar system for the progressive expansion of native bladder tissue. We investigated whether the expanded bladder tissue retained normal functional and phenotypic characteristics. MATERIALS AND METHODS: Urodynamic studies were performed in 5 beagle dogs and the bladder was divided horizontally into a superior bladder neo-reservoir, and an intact smaller bladder inferiorly with both ureters left intact and draining. A silicone catheter was threaded into the newly formed, superiorly located neo-reservoir, and connected to an injection port which was secured subcutaneously. A saline antibiotic solution was injected daily into the palpable injection port 4 weeks after surgery, dilating the neo-reservoir through the silicone catheter. Baseline and weekly cystograms were performed. Urodynamic studies of the neo-reservoirs were done immediately before sacrifice. Animals were sacrificed 3 months after the initial intervention and the bladder was examined grossly and microscopically. RESULTS: Within 30 days after progressive dilation, the neo reservoir volume was expanded at least 10-fold according to radiography and cystometrograms. Urodynamic studies of the dilated neo-reservoirs showed normal compliance in all animals. Microscopic examination of the expanded neo-reservoir tissue revealed normal histology. A series of immunocytochemical studies demonstrated that the dilated bladder tissue maintained normal phenotypic characteristics. CONCLUSIONS: The system of progressive dilation is effective in expanding bladder tissue which is able to retain normal phenotypic and functional characteristics. PMID- 10458391 TI - The hyaluronic acid receptor RHAMM is induced by stretch injury of rat bladder in vivo and influences smooth muscle cell contraction in vitro [corrected]. AB - PURPOSE: Loss of bladder compliance from hypercontractility and fibrosis may represent an injury response to excessive intravesical pressure. Together, interactions between cell and extracellular matrix components regulate cell response to injury and extracellular matrix remodeling. The receptor for hyaluronic acid mediated motility (RHAMM) is a recently described hyaluronic acid binding protein known to influence multiple types of cell extracellular matrix interaction in development, injury and cancer. We evaluate the role of RHAMM in mediating early events in bladder stretch injury. MATERIALS AND METHODS: An acute stretch injury model was used. The rat bladder was injured by hydrodistention inducing gross hematuria. Tissues were analyzed for temporal and spatial expression of RHAMM in the mucosa and detrusor regions by immunostaining, western and reverse transcriptase polymerase chain reaction analyses. The contractile activity of smooth muscle cell primary cultures was analyzed using a gel contraction assay in the presence of peptide fragments known to block RHAMM function. RESULTS: Acute hydrodistention caused immediate and significant injury to the bladder, with fracturing of smooth muscle cell bundles, edema and hemorrhage. RHAMM immunolocalized to the mucosa and detrusor within 2 hours of injury, peaking by 5 to 10 hours. A shift from low molecular weight (55 kD.) to high (120 kD.) receptor isoforms was prominent during the peak expression period noted by immunolocalization. RHAMM messenger ribonucleic acid increased only slightly (40%) by 5 hours after injury. Smooth muscle cell primary cultures actively initiated and maintained the contraction of collagen gels by more than 75% of baseline in vitro. Blocking RHAMM function significantly inhibited the ability to less than 25% of smooth muscle cells to contract the gels in vitro. CONCLUSIONS: Increased expression of RHAMM is an early event precipitated by stretch injury to the bladder. Since extracellular matrix hyaluronic acid is found early in tissue repair responses, its receptor RHAMM may be mediating initial bladder responses to stretch injury, some of which (contraction) may be experimentally blocked in vitro. Since the receptor directly regulates protein kinase signaling which in turn mediates smooth muscle cell contraction and collagen synthesis, further studies of RHAMM function in bladder pathology are warranted. PMID- 10458392 TI - Normal urodynamics in patients with bladder exstrophy: are they achievable? AB - PURPOSE: Urodynamic study was performed in patients with exstrophy to determine the effect of bladder neck reconstruction and the ability to achieve normal urodynamics following surgery. MATERIALS AND METHODS: A total of 30 exstrophy cases at different stages of reconstruction were retrospectively reviewed in terms of bladder capacity, compliance, stability and presence of detrusor contractions following urodynamic study. RESULTS: Bladder capacity increased from a third predicted volume for age to half after reconstruction. Approximately 80% of patients had compliant and stable bladders before bladder neck reconstruction. Following bladder neck reconstruction approximately half of the patients maintained normal compliance with a smaller number maintaining normal stability. A quarter of patients maintained normal filling dynamics following bladder neck reconstruction, and 19% maintained normal filling and voiding dynamics after reconstruction. CONCLUSIONS: The majority of closed exstrophy bladders have normal filling dynamics before bladder neck reconstruction. Compliance and stability are impaired following bladder neck reconstruction. Approximately 25% of patients with exstrophy may maintain normal detrusor function following reconstruction. However, less invasive alternatives to the Young-Dees-Leadbetter bladder neck reconstruction should be sought. PMID- 10458393 TI - The Mainz II pouch: experience in 5 patients with bladder exstrophy. AB - PURPOSE: We report our experience with 5 patients with bladder exstrophy who underwent creation of a Mainz II pouch. MATERIALS AND METHODS: We retrospectively reviewed the results of the Mainz II pouch as a primary urinary diversion in 2 and a secondary urinary diversion in 3 patients. Each patient underwent multiple surgeries, including osteotomy in 1. All patients were followed postoperatively on a yearly basis. RESULTS: All patients are continent and the upper urinary tract is stable. CONCLUSIONS: The Mainz II pouch is appropriate for children born with a small fibrotic bladder, and as a salvage procedure for those who have endured multiple reconstructive procedures and remain incontinent. Furthermore, this procedure deserves serious consideration in children residing in developing countries. PMID- 10458394 TI - Outcome of posterior urethral valves: to what extent is it improved by prenatal diagnosis? AB - PURPOSE: To assess the impact of prenatal diagnosis and evaluation on the outcome of posterior urethral valves we studied all cases of valves detected prenatally, including cases of pregnancy termination due to posterior urethral valves. MATERIALS AND METHODS: Between 1989 and 1996, 30 neonates with prenatally detected posterior urethral valves were treated at our hospital. The prenatal parameters analyzed were age of gestation at diagnosis, ultrasonographic appearance of renal parenchyma and amniotic fluid volume. Fetal urine was analyzed in 9 cases. We reviewed the outcome of 10 neonates treated for posterior urethral valves which were not diagnosed prenatally during the same period. RESULTS: Of the 30 neonatal survivors 6 (20%) had renal failure, including end stage renal disease in 2, after a mean followup of 4 years. Renal failure developed in 2 of 5 cases detected before 24 weeks of gestation, in 1 of 6 with oligohydramnios and in 2 of 5 with abnormal parenchymal renal ultrasound. Normal parenchymal ultrasound and amniotic volume could not predict for good outcome. Renal failure developed in 2 of 7 cases predicted by fetal urinalysis as good prognosis and in 1 of 2 cases predicted as poor prognosis. Pregnancy was terminated for posterior urethral valves in 5 cases based on prenatal criteria of severe renal impairment. Considering these cases as poor outcome, the rate of poor prognosis increased from 20 to 31%. Among the 10 neonates without a prenatal diagnosis of posterior urethral valves renal failure developed in 2 (20%), including end stage renal disease in 1. CONCLUSIONS: When negative parameters were absent and/or fetal urine predicted good outcome there were no cases of end stage renal disease in early infancy, which was a significant help in parent counseling. The predictive value of the currently available prenatal parameters needs to be updated with larger series specifically dealing with posterior urethral valves. According to the current data, the outcome of posterior urethral valves is not yet significantly improved by prenatal diagnosis. PMID- 10458395 TI - Fetal partial urethral obstruction causes renal fibrosis and is associated with proteolytic imbalance. AB - PURPOSE: We determine whether fetal bladder outlet obstruction induces renal fibrosis, and is associated with an alteration in the regulation of connective tissue degradation and the presence of fibrogenic interstitial cells. MATERIALS AND METHODS: Partial bladder outlet obstruction was surgically induced in 33 fetal sheep at 95 days of gestation. These animals and 24 normal age matched controls were sacrificed at 109, 116 and 135 (term) days of gestation, and the kidneys were rapidly retrieved, drained and weighed. Representative whole kidney samples were snap frozen for assessment of deoxyribonucleic acid, protein and collagen content. Morphometric analysis and alpha-smooth muscle actin immunohistochemistry were performed on histological specimens from formalin fixed kidneys. Tissue extract from fresh kidney specimens were analyzed for metalloproteinase and tissue inhibitor of metalloproteinase activity. Urine samples obtained at the time of sacrifice were analyzed for electrolyte, creatinine and N-acetyl glucosaminidase excretion. RESULTS: All obstructed kidneys were hydronephrotic and larger than age matched controls. Obstructed kidneys at term showed interstitial fibrosis, as measured by increased extracellular matrix volume fraction (45% in male obstructed kidneys versus 2.5% in normal male kidneys, p = 0.0004), increased total collagen content (120 mg./kidney in male obstructed versus 20 mg. in normal male animals, p = 0.016) and collagen/deoxyribonucleic acid content per kidney (2.78 versus 0.53 mg./mg., p = 0.016). Metalloproteinase-1 activity was significantly lower in obstructed kidneys (210 versus 380 U./mg. protein in normal kidneys). Tissue inhibitor of metalloproteinase activity was undetectable in both groups. The presence of an increased population of myofibroblasts often associated with fibrotic processes was seen by alpha-smooth muscle actin staining which was localized to interstitial cells throughout the cortex in obstructed kidneys. CONCLUSIONS: Fetal partial bladder outlet obstruction induces renal interstitial fibrosis as early as 2 weeks after obstruction. A possible mechanism for this process is a shift in proteolytic activity to reduce matrix degradation in obstructed kidneys. These changes might be mediated by the increased number of fibrogenic interstitial cells. The observations suggest several potential approaches to developing an understanding of congenital obstructive uropathy. PMID- 10458396 TI - Topical steroid treatment of phimosis in boys. AB - PURPOSE: We evaluate whether steroid application alone or retraction and hygiene are responsible for successful results in boys treated with topical steroids for phimosis. MATERIALS AND METHODS: A prospective study was performed, which included a control group of 42 patients with phimosis seen at our outpatient department from January to June 1997. During that time we trained the parent to retract and clean the foreskin only. From July 1997 to June 1998 topical steroid cream was prescribed in addition to retraction and hygiene in 276 boys with phimosis. All cases were divided into 3 subgroups of asymptomatic, symptomatic and buried penis. RESULTS: The response rate was greater than 95% in patients who received topical steroid treatment in addition to improved hygiene. Only 13 boys (less than 5%) had no response to steroid treatment. Of the control patients 23 (55%) had no response to gentle retraction and personal hygiene. There was a significant difference (p<0.001) in response rate between the study and control groups. However, the subgroup with a buried penis responded poorly to steroid, retraction and hygiene treatment. There was significant difference (p<0.001) in response rate between the buried penis and other steroid groups but no significant difference (p>0.05) in the control group. CONCLUSIONS: Phimosis is a physiological condition in neonates due to natural adhesion between the foreskin and the glans. Chronic infection due to poor hygiene is responsible for most cases of childhood phimosis. Circumcision is the traditional treatment of choice for phimosis or unretractable foreskin, although it is not always desired by parents or surgeons. Topical steroid cream is an easy, safe and nonsurgical alternative for phimosis. However, boys with a buried penis are not good candidates for steroid treatment. PMID- 10458397 TI - Effects of an environmental endocrine disruptor on fetal development, estrogen receptor(alpha) and epidermal growth factor receptor expression in the porcine male genital tract. AB - PURPOSE: We studied the effect of a potent reproductive tract toxin, 2,3,7,8 tetrachlorodibenzo-rho-dioxin, on fetal development and expression of estrogen receptor alpha and epidermal growth factor receptor (EGFR) in male swine. MATERIALS AND METHODS: Fetal domestic swine and miniswine were injected with 1 microg./kg. dioxin on day 50 of gestation and removed near term (114 days). Germ cell counts were performed on sections of formalin fixed testes. Estrogen receptor a protein, and messenger ribonucleic acid (mRNA) and EGFR mRNA expression were analyzed in frozen tissue using Western blotting and semiquantitative reverse transcriptase polymerase chain reaction. RESULTS: Of 15 dioxin exposed male offspring 8 (53%) had genital anomalies, including cryptorchidism in 4, epididymal detachment in 1, epididymal atresia in 1 and vasal dilatation in 3, while 3 of 17 control male swine (18%) had incompletely descended testes (p = 0.06). High intra-abdominal testes were found in 3 of 4 cryptorchid dioxin exposed but no control male swine. Mean germ cell number per tubule was 4.0+/-1.1 and 2.7+/-0.7 in control and dioxin groups, respectively (p = 0.01). Estrogen receptor a protein and mRNA were identified in fetal uterus, testis, gubernaculum and epididymis. Protein levels were 2 to 3-fold higher in dioxin exposed testis, and mRNA levels were significantly lower in gubernaculum and epididymis. EGFR mRNA expression was similar in treated and control testis and epididymis. CONCLUSIONS: Preliminary data suggest that dioxin produces cryptorchidism and wolffian duct anomalies in male swine exposed just before mid gestation. Germ cell counts and estrogen receptor alpha mRNA expression in gubernaculum and epididymis were significantly reduced, and estrogen receptor a protein expression in testis appeared to be increased by dioxin exposure. Aberrant regulation of estrogen receptor a expression by dioxin may contribute to reproductive tract anomalies in male fetuses. PMID- 10458398 TI - Germ cells may survive clipping and division of the spermatic vessels in surgery for intra-abdominal testes. AB - PURPOSE: Laparoscopy is a well described modality that provides an accurate visual diagnosis upon which further management of intra-abdominal testes may be based. Laparoscopic ligation of spermatic vessels as stage 1 of the procedure is a natural extension of laparoscopy. A staged approach provides adequate viability of the intra-abdominal testis. However, it is uncertain whether the more sensitive germ cells survive this procedure in addition to the Sertoli and interstitial cells of the human testis. Survival of germ cells is a prerequisite of later fertility potential. MATERIALS AND METHODS: We studied 17 nonpalpable testes in 10 patients 1 year and 7 months to 13(1/2) years old. Results of testicular biopsies of 13 intra-abdominal testes taken at stages 1 and 2 of surgery were available for histological comparison. RESULTS: Median number of spermatogonia per tubular cross section of the biopsies taken at stage 2 was slightly lower (0.03) compared to the median number at stage 1 (0.06) of the operation but this difference was not significant (p = 0.2031). CONCLUSIONS: Our study shows that the spermatogonia may survive clipping and division of the spermatic vessels, although the number of spermatogonia per tubular transverse section decreases slightly. PMID- 10458399 TI - Serum inhibin B levels and the response to gonadotropin stimulation test in pubertal boys with varicocele. AB - PURPOSE: The impact of varicoceles on gonadal function in adolescents has been evaluated using several parameters, including size of testes, hormonal levels and provocative endocrine testing. Inhibin B has been demonstrated to be decreased in men with testicular damage from conditions other than varicocele. We determine whether inhibin B levels are low in adolescent boys with varicocele, and if there is a relationship between inhibin B and an exaggerated response to gonadotropin releasing hormone (Gn-RH) stimulation testing, testicular hypertrophy and/or varicocele bilaterality. MATERIALS AND METHODS: We studied 9 boys at Tanner stages III to V of pubertal development who had either left or bilateral varicoceles. Basal inhibin B, follicle-stimulating hormone, luteinizing hormone and testosterone were measured. Each patient underwent Gn-RH stimulation testing. RESULTS: All patients had essentially normal inhibin B levels for Tanner stage. Of the 9 boys 4 had an exaggerated response to Gn-RH stimulating testing. Inhibin B levels did not vary significantly either with the presence of bilateral or unilateral varicoceles or asymmetric testis. CONCLUSIONS: The lack of correlation between inhibin B levels and the aforementioned parameters failed to suggest that inhibin B has a significant role in the clinical assessment of testicular function in adolescents with varicocele. Further studies of larger populations may further elucidate the value of inhibin B levels and varicoceles. PMID- 10458400 TI - Pathological difference between retractile and cryptorchid testes. AB - PURPOSE: We compared testicular biopsies from retractile and cryptorchid testes to determine the histological effect of testicular retraction and the necessity of treatment for retractile testes. MATERIALS AND METHODS: A total of 61 testicular biopsies were performed during orchiopexy in 36 boys 1.3 to 9.8 years old (mean age 5.4) with retractile testes (unilateral in 11, bilateral in 50) and 115 testicular biopsies were done in 83 patients with cryptorchidism (unilateral in 51, bilateral in 64) 0.5 to 14.9 years old (mean age 3.7). Parameters for germ and Sertoli cells were determined in each group. RESULTS: Mean average spermatogonial number (S/T value) and Sertoli cell index were statistically different between retractile and cryptorchid testes with values of 2.96+/-1.33 versus 0.61+/-0.87 and 26.81+/-6.75 versus 23.04+/-5.85, respectively. Average tubular degeneration phase V to VII ratio was 0.23+/-0.18 for retractile testes and 0.22+/-0.17 for cryptorchid testes which was not statistically different. CONCLUSIONS: Similar tubular degeneration phase V to VII values between retractile and cryptorchid testes indicate histological change in retractile testes and suggest the need for hormonal or surgical therapy for those patients with retractile testes lacking spontaneous descent. PMID- 10458401 TI - Paratesticular perineurioma: initial description. PMID- 10458402 TI - Antibiotic hydrogel coated Foley catheters for prevention of urinary tract infection in a rabbit model. AB - PURPOSE: We developed an antibiotic liposome (ciprofloxacin liposome) containing hydrogel for external coating of silicone Foley catheters and evaluated its efficacy in a rabbit model. Our goal was to create a catheter that would hinder the development of catheter associated nosocomial urinary tract infections. MATERIALS AND METHODS: We inserted either an untreated, liposomal hydrogel coated or a liposome hydrogel with ciprofloxacin coated 10F silicone Foley catheter into New Zealand White rabbits. We challenged the system with 5x10(6) virulent Escherichia coli at the urethral meatus twice daily for 3 days. Urine cultures were evaluated twice daily for 7 days. When urine cultures became positive, the rabbits were sacrificed and urine, urethral catheter and urethral tissue were cultured. RESULTS: The time to bacteriuria detection in 50% of the specimens was double for hydrogel with ciprofloxacin coated catheters versus untreated and hydrogel coated catheters. A significant (p = 0.04) improvement in average time to positive urine culture from 3.5 to 5.3 days and a 30% decrease in the bacteriuria rate for hydrogel with ciprofloxacin coated catheters were noted compared to untreated catheters. CONCLUSIONS: A significant benefit was realized by coating the extraluminal catheter surface with a ciprofloxacin liposome impregnated hydrogel. We believe this procedure will provide a significant clinical advantage, while reducing health care costs substantially. PMID- 10458403 TI - Differential expression of C-CAM cell adhesion molecule in prostate carcinogenesis in a transgenic mouse model. AB - PURPOSE: The transgenic adenocarcinoma of mouse prostate (TRAMP) model, in which various grades of prostate intraepithelial neoplasia (PIN) and prostate cancer with metastases can be reproducibly generated, is a paradigm for prostate disease progression. We have previously shown that C-CAM, an adhesion molecule, can suppress the growth of prostate cancer. In this report, we describe immunohistochemical characterization of differential expression of C-CAM at various stages of prostate tumorigenesis in the TRAMP model. MATERIALS AND METHODS: We sampled prostate specimens and periaortic lymph nodes from TRAMP mice. Indirect immunohistochemical staining with a polyclonal anti-C-CAM antibody was performed on the formalin-fixed, paraffin-embedded specimens. After castration at 12 weeks of age, the TRAMP mice developed androgen-independent prostate cancer (AIPC) and lymph node metastasis at 18 to 24 weeks of age. Samples from these castrated mice were also analyzed. RESULTS: C-CAM protein was expressed in the normal prostate epithelia of non-transgenic and TRAMP mice as well as in low-grade PINs in TRAMP mice. Expression was uniform on the luminal surfaces of these epithelia. C-CAM expression was noticeably reduced and the staining pattern heterogeneous in some high-grade PINs. C-CAM staining was generally absent in prostate cancer and metastatic lymph nodes. Androgen independent prostate cancer and its metastatic tumors generated in castrated TRAMP mice were also C-CAM negative. CONCLUSIONS: C-CAM expression correlates with the differentiation states of prostate epithelia and is down regulated early in prostate tumorigenesis in the TRAMP model. PMID- 10458404 TI - Clearance mechanism of prostate specific antigen and its complexes with alpha2 macroglobulin and alpha1-antichymotrypsin. AB - PURPOSE: To investigate the rate of elimination of prostate specific antigen (PSA) and its complexes with human alpha2-macroglobulin (alpha2-M) and alpha1 antichymotrypsin (ACT) and to elucidate the role of the alpha2-macroglobulin receptor/low density lipoprotein receptor-related protein (alpha2-M-R/LRP) in the clearance mechanism. MATERIALS AND METHODS: PSA and complexes of PSA with alpha2 M and ACT were prepared and radiolabeled with [125I]Na (Amersham, Braunschweig, Germany). Radiolabeled proteins were injected into rats and the elimination of radioactivity from circulation was measured by gamma-counting of 20 microL aliquots over time. After 30 minutes different organs were removed and the total radioactivity was counted. The elimination rate and distribution of PSA and PSA complexes was studied in the absence and presence of an excess of transformed alpha2-M. RESULTS: Radiolabeled PSA is rapidly eliminated from circulation with an initial half-life of 6.4+/-2.1 minutes mainly due to extraction by the liver and kidney. The clearance is slightly inhibited by transformed alpha2-M. PSA alpha2-M is solely eliminated by the liver with a half-life of 6.7+/-1 minutes. Uptake by the liver is competitively inhibited by transformed alpha2-M. PSA-ACT is eliminated by the liver and kidney with an initial half-life of 3.51+/-1.1 minutes. Transformed alpha2-M failed to inhibit the clearance of PSA-ACT. CONCLUSIONS: Free PSA and PSA-inhibitor complexes are removed from the circulation by different clearance mechanisms. The sites of metabolism of the different forms of PSA are different but include liver and kidney as main organs for uptake. There are indications that alpha2-M-R/LRP is involved in PSA elimination. Thus, factors which modulate the receptor function and expression as well as the concentration of its natural ligands may interfere with the steady state concentrations of different PSA forms in blood. PMID- 10458405 TI - Simple anti-reflux uretero-ileal anastomosis: an experimental study in dogs. AB - PURPOSE: To develop more simple and effective anti-reflux techniques applicable to the ileal reservoir, we examined the usefulness of a novel anti-reflux uretero ileal anastomosis creating a flap valve mechanism. MATERIALS AND METHODS: Five dogs were used. A 4 cm. long ileal segment was isolated and its oral part (2.5 cm.) was detubularized. The detubularized ileal plate was anastomosed to the dome of the bladder. The left ureter was cut and its proximal end was spatulated and anastomosed end-to-side to the ileal plate. The end of the 1.5 cm. long afferent limb (non-detubularized part of the ileal segment) was closed and fixed down to the ileal plate to create an extramural tunnel. In this way, the reimplanted ureter was covered by the afferent limb. The dogs were evaluated between 3 and 4 months postoperatively. RESULTS: None of the 5 dogs used showed vesico-ureteral reflux or hydronephrosis. Histological examination showed an intact ureter enclosed with a normal ileal wall. CONCLUSION: Our proposed anti-reflux uretero ileal anastomosis is simple and reliable. This technique may be suitable for applying to a urinary reservoir, especially as an alternative to the intussuscepted nipple valve. PMID- 10458406 TI - Expression of messenger RNAs for membrane-type 1, 2, and 3 matrix metalloproteinases in human renal cell carcinomas. AB - PURPOSE: Three different membrane-type matrix metalloproteinases (MT1, 2, and 3 MMP) that can activate proMMP-2 (progelatinase A) are thought to play an important role in invasion and metastasis by various human carcinomas. To further clarify this role, we examined mRNA expression of MT-MMPs in human renal cell carcinomas. MATERIALS AND METHODS: mRNA was extracted from 25 clinical specimens of renal cell carcinoma and 23 specimens of normal renal parenchyma remote from the tumor. Reverse transcription polymerase chain reaction (RT-PCR) using specific primers was performed, and PCR products were hybridized to 32P-labeled internal probes and analyzed by a bioimage analyzer. RESULTS: MT1, 2, and 3-MMP mRNA expression in carcinomas was significantly higher than in normal parenchyma. In terms of the pathologic stage, MT1-MMP mRNA expression in pT2 and pT3 tumors was significantly higher than those in pT1 tumors. Although the sample size was small, it was evident that MT3-MMP mRNA expression in clear cell subtype renal cell carcinomas was higher than in the group of tumors including the granular cell subtype. CONCLUSIONS: These three MT-MMPs may play an important role in the pathogenesis of human renal cell carcinoma, and MT1-MMP in particular is important in invasion by carcinoma cells. It is interesting that the expression of MT3-MMP was higher in carcinomas, especially clear cell carcinoma, than in normal parenchyma, so that MT3-MMP may provide a clue an understanding of the molecular mechanism of carcinogenesis in human kidney. PMID- 10458407 TI - Transforming growth factor-beta1 (TGF-beta1) is sufficient to induce fibrosis of rabbit corpus cavernosum in vivo. AB - PURPOSE: The pleotropic cytokine TGF-beta1 which induces connective tissue synthesis, and inhibits the growth of smooth muscle cells, has been implicated in corpus cavernosum fibrosis. The objective of this study was to determine the dose and time dependence of TGF-beta1 as an active agent in penile corporal fibrosis in an animal model. MATERIALS AND METHODS: A time release method of delivery was developed using sodium alginate microspheres containing recombinant human (rh) TGF-beta1. New Zealand White rabbits were injected intracorporally with a single alginate microsphere either with or without rh-TGF-beta1. Dosage was varied from 325 to 1500 ng./bead. Animals were sacrificed at either three or five days post injection and the penises removed en bloc, examined, and processed for quantitative histomorphometric analysis, staining the sections with Masson's trichrome. RESULTS: Alginate microspheres containing [125I]-rh-TGF-beta1 showed slow-release kinetics (t1/2 = 10.5 hours). Histomorphometric analysis of 60 sets of high powered fields/treatment/ animal showed dose dependent decreases in percentage of corporal smooth muscle with TGF-beta1 treatment (750 to 1500 ng./bead). Placebo (alginate microspheres alone) had trabecular smooth muscle content comparable to values previously reported for untreated rabbit corpus cavernosum. CONCLUSIONS: This study confirms that TGF-beta1 induces fibrosis in situ by altering connective tissue synthesis and hence the structure of the corpus cavernosum. Injection of rh-TGF-beta1 impregnated alginate microspheres into the corpus cavernosum resulted in dose-dependent decreases in percentage of corporal smooth muscle. PMID- 10458408 TI - Calcium ionophore, ionomycin inhibits growth of human bladder cancer cells both in vitro and in vivo with alteration of Bcl-2 and Bax expression levels. AB - PURPOSE: The objective of this study was to characterize the antiproliferative effects of the calcium ionophore, ionomycin on the human bladder cancer cell line HT1376 both in vitro and in vivo. MATERIALS AND METHODS: The in vitro growth rate of HT1376 after treatment with ionomycin was measured by using the MTT assay. The apoptotic features in HT1376 after ionomycin treatment were evaluated by flow cytometric analysis and DNA degradation assay. Bcl-2 and Bax expression levels in HT1376 after ionomycin treatment were examined by Northern and Western blot analyses. The effects of intratumoral injection of ionomycin on the growth of subcutaneous HT1376 tumors established in athymic nude mice were then tested. The efficacy of combined treatment with ionomycin and cisplatin against HT1376 growth was also examined. RESULTS: The in vitro growth rate of HT1376 was suppressed by ionomycin in a dose- and time-dependent manner, and characteristic apoptotic DNA degradation was observed in HT1376. Ionomycin treatment caused a marked decrease in the ratios of Bcl-2 to Bax mRNA and protein in HT1376 cells. Intratumoral injection of ionomycin into subcutaneous HT1376 tumors reduced the tumorigenicity in nude mice. Furthermore, these in vivo growth-inhibitory effects of ionomycin were significantly enhanced by pretreatment with cisplatin. CONCLUSIONS: These findings suggest that ionomycin-based therapy could be used as a novel therapeutic strategy for advanced bladder cancer through the effective induction of apoptosis by decreasing the ratio of Bcl-2 to Bax. PMID- 10458409 TI - Loss of heterozygosity on chromosome 13 is associated with advanced stage prostate cancer. AB - PURPOSE: In order to investigate the possible involvement of a tumor suppressor gene(s) on chromosome 13 in prostatic neoplasms, we performed loss of heterozygosity (LOH) analysis on normal and tumor pairs from 36 prostate cancer patients. MATERIALS AND METHODS: Pure DNA was obtained from carcinoma cells and normal epithelium by tissue microdissection. The DNA had previously been analyzed for LOH on chromosomes 8 and 16. After an initial pilot experiment to determine the region(s) of significant LOH from 9 loci on chromosome 13q, 3 loci at and near the Rb1 locus (D13S153, D13S1319, and D13S1303) were chosen for further study. RESULTS: The overall rate of LOH on chromosome 13 was 27.3%. Four tumors exhibited LOH at all 3 loci. Two tumors exhibited LOH at D13S153 but not at the other, more telomeric loci; two additional tumors had loss at D13S1303 or D13S1319 but not D13S153. These data suggest that a tumor suppressor gene involved in prostate cancer may be located just telomeric to Rb1. Analysis of clinical and pathological data from carcinomas with and without loss shows that chromosome 13q LOH is correlated with advanced stage prostate cancer. CONCLUSIONS: Our LOH data suggests that there may be a tumor suppressor gene telomeric to Rb1 that is potentially involved in prostate cancer progression. Identification of this gene may be valuable in providing diagnostic and prognostic information for prostate cancer patients. PMID- 10458410 TI - Differential RNA expression of the pS2 gene in the human benign and malignant prostatic tissue. AB - PURPOSE: The pS2 trefoil protein has been detected in close association with neuro-endocrine differentiation in prostate cancer and prostatic intraepithelial neoplasia. These preliminary results have suggested that pS2 is a candidate as a specific marker for prostate cancer tissue. To ascertain the specificity of pS2 in prostate cancer tissue, we have used an RT-PCR method from prostate biopsies provided from human malignant and benign prostatic hyperplasia (BPH) tissue. MATERIALS AND METHODS: Prostate biopsies were obtained from transrectal biopsies from 153 patients with an abnormal DRE or a PSA more than 4 ng./ml. or symptoms of BPH and a PSA more than 4 ng./ml. Total RNA was extracted from fresh frozen specimens of tissue samples. Detection of pS2 transcript compared with GADPH transcripts was done using RT-PCR. RESULTS: Biopsy results showed that 108 patients had prostate cancer (average Gleason score 6.39+/-0.74) and 45 patients had BPH. PS2 RT-PCR results showed that PS2 RNA expression was negative in 83% of the BPH cases. Conversely, 92% of prostate cancer specimens were positive (Chi square: 86.09, p<0.001). There was no correlation with tumor stage or the Gleason score. Comparing the expression of pS2 in BPH and localized prostate cancer, we found a sensitivity of 92% and a specificity of 82%. CONCLUSIONS: On this large sample of prostate biopsies from patients at risk of having prostate cancer, pS2 was demonstrated as an interesting marker significantly associated with prostate cancer. Further work on the expression of pS2 according to differentiation and hormonal status is in progress. PMID- 10458411 TI - Detection of circulating uroplakin-positive cells in patients with transitional cell carcinoma of the bladder. AB - PURPOSE: Although transitional cell carcinoma of the bladder (TCC) metastasizes frequently with devastating consequences, no marker has been available to monitor this process. Uroplakins are a group of specific markers for normal urothelium and are continuously expressed by the majority of TCCs. Detection of uroplakin positive cells in the circulation would be a strong indication of hematogenous dissemination of tumor cells in patients with TCC. MATERIALS AND METHODS: Total RNAs were extracted from peripheral blood of 60 patients with TCC (50 non metastatic and 10 metastatic) and 10 healthy controls, reverse-transcribed and subjected to polymerase chain reaction amplification (RT-PCR) using oligonucleotide primers of human uroplakin II gene. A uroplakin-expressing human bladder cancer cell line (RT4) was used as a positive control to establish the sensitivity of the RT-PCR assay. RESULTS: We showed that the PCR-amplification of the mRNA encoding uroplakin II (UPII), a 15-kDa urothelium-specific marker, constitutes a highly sensitive and specific assay for detecting 100% of transitional cell carcinoma tissue, and that this assay can detect a single bladder cancer cell in a 5-ml. blood sample. UPII mRNA was detected in the blood samples of 2 patients with metastatic bladder cancer without chemotherapy and 1 out of 8 such patients with chemotherapy, but not in those of 50 non-metastatic patients or normal controls. CONCLUSIONS: Uroplakin II is a highly specific marker for human TCC and the detection of uroplakin II in the peripheral blood is associated with metastatic spread of bladder cancer cells. The specific and sensitive detection of uroplakin II provides a useful adjunct for detecting bladder cancer metastasis, staging, and monitoring chemotherapeutic response. PMID- 10458412 TI - "Skin-CNS-bladder" reflex pathway for micturition after spinal cord injury and its underlying mechanisms. AB - PURPOSE: A "skin-CNS-bladder" reflex pathway for inducing micturition after spinal cord injury has been established in cat. This reflex pathway which is basically a somatic reflex arc with a modified efferent limb that passes somatic motor impulses to the bladder, has been designed to allow spinal cord injured patients to initiate voiding by scratching the skin. MATERIALS AND METHODS: The skin-CNS-bladder reflex was established in the cat by intradural microanastomosis of the left L7 ventral root (VR) to the S1 VR while leaving the L7 dorsal root (DR) intact to conduct cutaneous afferent signals that can trigger the new micturition reflex arc. After allowing 11 weeks for axonal regeneration, urodynamic, pharmacological and electrophysiological studies were conducted in pentobarbital or chloralose anesthetized animals. RESULTS: A detrusor contraction was initiated at short latency by scratching the skin or by percutaneous electrical stimulation in the L7 dermatome. Maximal bladder pressures during this stimulation were similar to those activated by bladder distension in control animals. Electrophysiological recording revealed that single stimuli (0.3 to 3 mA, 0.02 to 0.2 msec duration) to the left L7 spinal nerve in which the efferent axons had degenerated evoked action potentials (0.5 to 1 mV) in the left S1 spinal nerve distal to the anastomosis. In addition, increases in bladder pressure were elicited by trains of the stimuli (5 to 20 Hz, 5 seconds) applied to the L7 spinal nerve. Urodynamic studies including external sphincter EMG recording demonstrated that the new reflex pathway could initiate voiding without detrusor-external urethral sphincter dyssynergia. Atropine (0.05 mg./kg., i.v.) or trimethaphan (5 mg./kg., i.v.), a ganglionic blocking agent, depressed the bladder contractions elicited by skin stimulation. The skin-CNS-bladder reflex could also be elicited after transecting the spinal cord at the L2-L3 or L7-S1 levels. CONCLUSION: The cross-wired somato-autonomic bladder reflex is effective in initiating bladder contractions and coordinated voiding in cats with an intact neuraxis and can also induce bladder contractions after acute transection of the lumbar spinal cord. The new pathway is mediated by cholinergic transmission involving both nicotinic and muscarinic receptors. It is concluded that somatic motor axons can innervate bladder parasympathetic ganglion cells and thereby transfer somatic reflex activity to the bladder smooth muscle. PMID- 10458413 TI - Effects of intravesical administration of the K+ channel opener, ZD6169, in conscious rats with and without bladder outflow obstruction. AB - PURPOSE: To investigate the urodynamic effects of the new K(ATP) channel opener, ZD6169, given intravesically, in rats with and without bladder outflow obstruction. MATERIALS AND METHODS: Female, conscious Sprague-Dawley rats, normal or with bladder hypertrophy and overactivity secondary to bladder outflow obstruction, were given ZD6169 intravesically (10 or 100 ng./ml.), and intra arterially (1 mg./kg.). Continuous cystometry was performed. RESULTS: In normal and obstructed rats, intravesical ZD6169 had similar, dose-dependent effects on bladder function. In obstructed rats, ZD6169 (100 ng./ml.) significantly decreased micturition pressure (17%), and increased bladder capacity (32%), micturition volume (18%), residual volume (145%), and inter-contraction interval (71%). There was a marked decrease in both frequency (40%) and amplitude (43%) of the spontaneous bladder activity. When given intra-arterially in obstructed rats ZD6169 increased bladder capacity (19%) and residual volume (47%) and decreased amplitude (51%), but not frequency, of the spontaneous bladder activity. CONCLUSIONS: In both normal and obstructed rats, intravesical ZD6169, at the investigated doses, significantly affected bladder function. In obstructed rats, the drug markedly reduced bladder overactivity. If the results have clinical validity, intravesical ZD6169 may offer an alternative way of treating bladder overactivity in selected patients. PMID- 10458415 TI - A comparison of recipient renal outcomes with laparoscopic versus open live donor nephrectomy. PMID- 10458414 TI - Cytotoxicity of adenoviral-mediated cytosine deaminase plus 5-fluorocytosine gene therapy is superior to thymidine kinase plus acyclovir in a human renal cell carcinoma model. AB - PURPOSE: An estimated 11,600 Americans will die of renal cell carcinoma in 1998. The lack of effective chemotherapy or radiotherapy requires the investigation of novel treatment modalities. We compared two forms of toxic gene therapy, cytosine deaminase (CD) plus 5-fluorocytosine (5-FC) and thymidine kinase (TK) plus acyclovir (ACV), in pre-clinical models of human renal cell carcinoma. MATERIALS AND METHODS: Replication-deficient recombinant adenoviral vectors containing the Rous sarcoma virus promoter driving CD (Ad-RSV-CD) or TK (Ad-RSV-TK) gene expression were constructed and tested for in vitro cell-killing assays at various viral multiplicity of infection (MOI) and in vivo for growth inhibition of a human renal cell carcinoma, SK-RC-29 models. Subcutaneous tumors of SK-RC-29 were examined by electron microscopy for presence of intercellular gap junctions. Levels of expression of the gap junctional associated connexin 43 protein in SK RC-29, 31, 38, 42, 52 human RCC cell lines were examined by western immunoblotting. RESULTS: In vitro cell-killing assay comparing Ad-RSV-CD/5F-C and Ad-RSV-TK/ACV at a wide range of MOI (2.5 to 20) revealed superior cell-kill by Ad-RSV-CD/5-FC over Ad-RSV-TK/ACV. Consistent with these results, we observed that Ad-RSV-CD/5-FC but not Ad-RSV-TK/ACV demonstrated a significant in vivo tumor growth inhibition. These results are corroborated by the lack of gap junctions in SK-RC-29 subcutaneous tumors by the electron microscopy and the absence of connexin-43 in all five human RCC cell lines by western immunoblotting. CONCLUSION: We have demonstrated in this study that Ad-RSV-CD/5 FC is superior to Ad-RSV-TK/ACV for the treatment of human RCC in cell culture and animal models. The results are supported by the lack of gap junctional communication between RCC cells assessed by connexin-43 expression. PMID- 10458416 TI - An unusual subset of cryptorchidism: possible end organ failure. AB - PURPOSE: A paucity of germ cells exists in the cryptorchid gonad that usually correlates with a similar finding in the contralateral descended testis. However, we have noted a small number of boys with cryptorchidism in whom there is a significant difference between the histological evaluation of the cryptorchid testis and the normal descended testis that may indicate a different etiology. MATERIALS AND METHODS: From 1986 to 1991, 1,426 boys with unilateral cryptorchidism underwent orchiopexy, of whom 752 also underwent bilateral testicular biopsy. Testicular volume and position, and patency of the processus vaginalis were examined. Biopsies were fixed in 2% glutaraldehyde and embedded in Epon. Semithin tissue sections were analyzed by 2 independent investigators. The number of total germ cells, gonocytes, adult dark and pale spermatogonia, primary spermatocytes and Leydig cells was assessed. RESULTS: Of the 1,426 boys the undescended testis was on the right side in 726 and on the left side in 658 (52 versus 48%, p = 0068). Of the 752 boys who underwent bilateral biopsy 42 (5.6%) 1.1 to 16 years old (mean age plus or minus standard deviation 0 5.2 +/- 3.65) had a poor fertility index of less than 0.2 germ cell per tubule in the cryptorchid gonad, although the germ cell count in the descended testis was normal. Of the 42 testes in this special group of boys 30 (71%) were on the right side (Fisher's exact test p <0.23), including 16 (38%) in an intra-abdominal or high canalicular position. The processus vaginalis was patent in 86% of the intra abdominal testes and in 100% of those located at the tubercle but in only 25% of those in a pre-scrotal position. While average germ cell count in the cryptorchid testis was 0.06 per tubule with abnormal germ cell maturation, number was normal (greater than 2 germ cells per tubule) in the contralateral descended testis with a normal distribution of adult dark and pale spermatogonia, and primary spermatocytes. Average volume of the cryptorchid testis was significantly less than that of the descended testis (1.20 +/- 0.35 versus 1.60 +/- 0.68 mm.3, p <0.0001). CONCLUSIONS: Based on the normal scrotal testis the fertility prognosis is good in this small subgroup of boys with cryptorchidism. Rather than the usual endocrinopathy of cryptorchidism, the undescended testis in these boys may be the result of end organ failure. These patients with favorable fertility potential may be recognized only if each testis is biopsied at unilateral orchiopexy. PMID- 10458417 TI - Age at unilateral orchiopexy: effect on hormone levels and sperm count in adulthood. AB - PURPOSE: We determined whether there are differences in hormone levels, such as inhibin B, follicle-stimulating hormone (FSH), luteinizing hormone and testosterone, and sperm density in men with a history of unilateral cryptorchidism as stratified by age at orchiopexy. MATERIALS AND METHODS: A total of 84 men with a history of unilateral cryptorchidism presented to our institution for serum hormone measurement and semen analysis. These parameters were compared using Pearson's correlations and analysis of variance among 4 groups stratified according to age at orchiopexy (range 1 month to 11 years). RESULTS: Comparison by Pearson's correlation analysis showed that age at orchiopexy significantly correlated inversely with inhibin B (r = -0.274, p = 0.012) and positively correlated with FSH (r = 0.229, p = 0.036). Comparison of mean hormone levels and sperm density by analysis of variance for linear trend revealed a significant relationship between age at surgery with inhibin B (p = 0.032) and testosterone (p = 0.029), while sperm density, FSH and luteinizing hormone were not significantly related. Post hoc comparison of individual means at surgery and at the time of this study demonstrated a significantly higher inhibin B level in the youngest age group than in 2 of the 3 older groups. CONCLUSIONS: Men who previously had unilateral cryptorchidism and who underwent orchiopexy by age 2 years have higher inhibin B and lower FSH profiles than those who underwent surgery later in life. This finding suggests an overall beneficial effect of early orchiopexy in boys born with unilateral cryptorchidism. PMID- 10458418 TI - Laparoscopic Fowler-Stephens orchiopexy for the high abdominal testis. AB - PURPOSE: Laparoscopic orchiopexy is extremely effective for treating patients with nonpalpable testis. However, despite the high dissection and wide mobilization it allows in some cases, vessel length prevents the testis from reaching the scrotum. There have been only incidental cases reported in which laparoscopy has been used for vessel transection and testicular mobilization orchiopexy. We reviewed our cases treated with the Fowler-Stephens orchiopexy performed laparoscopically in 1 or 2 stages. MATERIALS AND METHODS: We reviewed the records of all boys who underwent laparoscopy for a nonpalpable testis at our institutions since 1992. Patients who underwent testicular vessel transection and orchiopexy performed laparoscopically in 1 or 2 stages were selected for evaluation. Office charts and operative reports were reviewed in detail. RESULTS: Of the 126 nonpalpable testes in 108 patients 51 (40%) were intra-abdominal, including 18 (35%) in 14 patients in whom the Fowler-Stephens procedure was performed laparoscopically. Five testes were treated with a 2-stage procedure, while 11 were managed by laparoscopic mobilization followed by laparoscopic vessel clipping and orchiopexy in 1 stage. In 2 additional patients nearly all dissection was performed laparoscopically but due to extenuating circumstances inguinal incision was required as well. Thus, 13 testes were managed by 1-stage Fowler-Stephens orchiopexy, including all cases since August 1996 which required vessel transection. Two patients were hospitalized postoperatively for prolonged ileus after the second stage. All other 2-stage and all 1-stage cases were managed on an outpatient basis. There were no complications. At a mean followup of 6 months all cases without previous surgery that were managed by laparoscopic orchiopexy are without atrophy and the testes are in a scrotal position. Two testes in which previous surgery had been done atrophied postoperatively. CONCLUSIONS: Laparoscopic transection of the testicular vessels is safe in boys with high abdominal testes that do not reach the scrotum after laparoscopic high retroperitoneal dissection. The magnification and wide mobilization of laparoscopy likely allow better preservation of the collateral vascular supply than open exploration. Previous surgery is a risk factor for atrophy. The success rate of 89% overall and 100% in patients who did not previously undergo testicular surgery equals or exceeds that of open orchiopexy in patients with abdominal testes. The 1-stage procedure avoids repeat anesthesia and the extensive, sometimes tedious, dissection that is occasionally required during reoperation. PMID- 10458419 TI - Is preoperative laparoscopy useful for impalpable testis? AB - PURPOSE: Laparoscopic assessment of pelvic anatomy has gained wide popularity over the years. Today surgical treatment of impalpable testis is nearly always preceded by diagnostic laparoscopy. The actual role of such a procedure remains undefined. We performed a prospective randomized clinical trial in patients with impalpable testis to evaluate the clinical usefulness of laparoscopy before surgical exploration. MATERIALS AND METHODS: We studied pediatric, age matched patients with impalpable testis who were randomized to group 1-30 who underwent open surgery only and group 2-31 who underwent laparoscopy and open surgery. Anatomical findings, operative procedures, operative time and cost, number of recurrences and testicular volume at followup were then compared in the 2 groups. RESULTS: There were no statistically significant differences in the 2 groups for any of the considered parameters except operative cost and time, which were significantly higher in the laparoscopy group. CONCLUSIONS: Preoperative laparoscopy does not provide any significant advantage over open surgery for treating impalpable testis. PMID- 10458420 TI - A new management algorithm for impalpable undescended testis with gadolinium enhanced magnetic resonance angiography. AB - PURPOSE: We evaluated the diagnostic accuracy of preoperative localization of impalpable undescended testis using ultrasound and gadolinium (Gd) enhanced magnetic resonance angiography (MRA). MATERIALS AND METHODS: Ultrasound and Gd MRA were performed prospectively in 21 boys (23 impalpable testes) with cryptorchidism before laparoscopy and surgical exploration. Gd-MRA was done using a 1.5 Tesla magnetic resonance scanner with a turbo field echo technique after bolus intravenous injection of 0.4 mmol./kg. body weight of Gd diethylenetriaminepentaacetic acid. A total of 10 dynamic scans were acquired during 15 minutes after Gd injection to cover the early arterial and delayed venous phases. All patients subsequently underwent diagnostic laparoscopy and definitive surgery. RESULTS: Ultrasound correctly localized 9 of 10 intracanalicular testes but failed to reveal any intra-abdominal or vanishing testes. In contrast, Gd-MRA correctly localized 4 intra-abdominal, all 10 intracanalicular and 8 canalicular vanishing testes. In 1 patient with an intra abdominal vanishing testis ultrasound and Gd-MRA failed to make the diagnosis. When correlated with the findings of subsequent laparoscopy and surgical exploration, Gd-MRA had a diagnostic sensitivity of 96% and a specificity of 100% for localizing impalpable undescended testes. Based on Gd-MRA and ultrasound findings laparoscopy could have been avoided in 18 of 23 cases (78%). No adverse effect was associated with Gd use in this study. CONCLUSIONS: Gd-MRA accurately diagnoses vanishing testes and reliably differentiates intraabdominal from intracanalicular impalpable testes, allowing definite preoperative planning of the surgical approach and avoiding unnecessary laparoscopy. A new management algorithm for impalpable testis in patients with cryptorchidism is proposed based on ultrasound and Gd-MRA findings. PMID- 10458421 TI - The incidence of intersexuality in children with cryptorchidism and hypospadias: stratification based on gonadal palpability and meatal position. AB - PURPOSE: The combined findings of cryptorchidism and hypospadias often indicate the existence of an intersex state. Testicular maldescent and incomplete tubularization of the urethral plate occur in a spectrum with the severity of the 2 processes likely dependent on the degree of pathophysiology in the androgenic hormonal axis. The incidence of intersexuality in children with undescended testes, hypospadias and otherwise nonambiguous male genitalia has been reported to be 27%. Although the likelihood of genotypic or gonadal ambiguity has previously been associated with meatal position in this population, to our knowledge our study is the first to evaluate the incidence of intersexuality relative to whether the undescended testis is palpable or nonpalpable. MATERIALS AND METHODS: The database at our hospital was searched for all cases of undescended testes (2,105) and hypospadias (1,057) between 1982 and 1996. Radiographic, histological and karyotypic data were compiled for all patients presenting with both diagnoses. Gonadal palpability (palpable versus nonpalpable) and meatal position (anterior versus mid versus posterior) were recorded and correlated with the likelihood of identifying an intersex condition. Ten boys with a diagnosis of undescended testes subsequent to inguinal hernial repair were excluded from study. Patients with congenital adrenal hyperplasia or complete testicular feminization were also excluded from study due to the clearly female appearance of the external genitalia. Statistical significance was assessed using Fisher's exact test. RESULTS: We identified 79 patients presenting with undescended testes, hypospadias and a phallus that was believed to be a possible penis. Intersex conditions were identified with nearly equal frequency in the 44 cases of unilateral (30%) and 35 of bilateral (32%) cryptorchidism. In the unilateral undescended testes group patients with a nonpalpable testis were at least 3-fold more likely to have an intersex condition than those with a palpable undescended testis (50 versus 15%, p = 0.02). In the bilateral group patients with 1 or more nonpalpable testes were also nearly 3-fold as likely to have an intersex condition than those with bilateral palpable undescended gonads (47 versus 16%, p = 0.07). Meatal position was graded as anterior in 33% of cases, mid in 25% and posterior in 41% with the more posterior location conferring a significantly greater likelihood of an intersex condition (anterior 2 of 26, mid 1 of 20 and posterior 21 of 33). CONCLUSIONS: Gonadal palpability is an important predictor of an intersex state in unilateral and bilateral cases of cryptorchidism with hypospadias. Patients with an undescended testis that cannot be palpated are significantly more likely to have an intersex condition than those in whom the undescended testis may be palpated on physical examination. The severity of hypospadias likewise has a strong positive correlation with an intersex state. PMID- 10458422 TI - Mullerian duct remnants: surgical management and fertility issues. AB - PURPOSE: We reviewed our experience with mullerian duct remnants, also known as prostatic utricular and mullerian duct cysts, to advance further the understanding of the surgical management of these challenging congenital anomalies. The indications, merits and disadvantages of each surgical approach are presented, and the effects of mullerian duct remnants and their treatment on future fertility are discussed. MATERIALS AND METHODS: We reviewed the records of 26 patients 1 month to 19 years old with mullerian duct remnants who were seen between January 1984 and October 1998. Clinical presentation included perineoscrotal hypospadias in 10 cases, urinary retention and/or difficult voiding in 7, urinary tract infection in 6, acute scrotum in 2, and recurrent hemospermia and dysuria in 1. RESULTS: Of the 26 patients 13 required surgical intervention for various symptoms and to correct large diverticula. The surgical approach was transvesical transtrigonal in 8 cases, extravesical in 2, perineal in 2 and posterior sagittal in 1. Transurethral fulguration was performed in 2 cases. The initial surgical approach was successful in 11 of the 13 patients. One patient required conversion to a transvesical transtrigonal approach due to inadequate exposure during attempted perineal excision. Two cases treated with transurethral fulguration failed to resolve completely, and in 1 excision was required using the transvesical transtrigonal technique. A total of 13 patients were treated nonoperatively, including 10 in whom the condition was discovered incidentally during screening for perineoscrotal hypospadias. In 5 of the 10 patients urinary tract infection subsequently developed and they were maintained on long-term chemoprophylaxis. CONCLUSIONS: By tailoring the surgical approach to the type of mullerian duct remnant and the relevant anatomical relationships a high degree of success may be achieved with minimal morbidity. PMID- 10458423 TI - Anatomical studies of the human clitoris. AB - PURPOSE: Endogenous or exogenous testosterone exposure to the fetus during gestation may result in masculinization of the external genitalia. Surgical correction requires a clear understanding of normal female anatomy. We report observations from our anatomical dissections on which we base our approach to reduction clitoroplasty. MATERIALS AND METHODS: A total of 14 normal human fetal clitoral specimens at 8 to 24 weeks of gestation were serially sectioned after formalin fixation. Every tenth section was stained with Masson's trichrome, smooth muscle alpha-actin and the neuronal markers PGP 9.5 or S-100. Computer reconstruction using imaging software permitted 3-dimensional analysis of the nerves, corporeal bodies and glans clitoris. These specimens were compared with 2 obtained postnatally at feminizing genitoplasty. RESULTS: The normal fetal clitoris consists of 2 corporeal bodies with a midline septum. The ultrastructure of the female corporeal bodies is similar to that of the male counterpart. The glans clitoris forms a cap on top of the distal end of the narrowed corporeal bodies. There is a midline septum starting on the ventral aspect and extending approximately halfway into the glans. Large bundles of nerves course along the corporeal bodies with the highest density on the dorsal aspect or top. No nerves were noted at the 12 o'clock position, although nerves extend completely around the tunica in a fashion similar to that of the fetal penis. Glans innervation is provided by multiple perforating branches entering at the dorsal junction of the corporeal body and glans. The lowest density of nerves in the glans is on the ventral aspect in juxtaposition to the glans septum. In surgical specimens obtained from patients with congenital adrenal hyperplasia nerves were adjacent to the excised tunica of the corporeal bodies, especially on the lateral aspect. CONCLUSIONS: A clear understanding of the anatomy of the human clitoris is important for surgical reconstruction. As in the human penis, nerves in the clitoris form an extensive network around the tunica of the corporeal body with a nerve-free zone at the midline 12 o'clock position. Care should be taken to preserve all nerves. Reduction of the glans clitoris should not violate the extensive innervation that predominates on the dorsal aspect of the glans. The normal clitoris has corporeal bodies that are smaller but analogous to those of the penis. One may consider their function when extensive resection is considered. PMID- 10458424 TI - Pediatric management of ambiguous and traumatized genitalia. AB - PURPOSE: The present standard of practice in the management of ambiguous and traumatized genitalia was evaluated. MATERIALS AND METHODS: Published cases of intersexuality and protocols for the management of traumatized genitalia were reviewed with consideration of the input of intersexual individuals. Independent research on different types of intersexuality is also presented. RESULTS: The present standard pediatric recommendations and precepts for the management of ambiguous or traumatized genitalia are wanting. Followup studies on which to base treatment decisions are needed. Evidence based principles of medical management are proposed. CONCLUSIONS: A moratorium on sex reassignment cosmetic surgery is recommended. Also recommended are that followup studies should be instituted on past cases, and honesty and counseling should be the core of initial and subsequent treatment. PMID- 10458425 TI - Prenatal diagnosis of hydronephrosis: impact on renal function and its recovery after pyeloplasty. AB - PURPOSE: We reviewed our experience with corrective surgery for congenital ureteropelvic junction obstruction to assess the impact of mode of presentation on renal function at diagnosis and on postoperative recovery of function. MATERIALS AND METHODS: We reviewed the records of consecutive children who underwent pyeloplasty or nephrectomy for ureteropelvic junction obstruction during a 5-year period at our hospitals. Patients were divided into those with and without a prenatal diagnosis of hydronephrosis. In each group we compared preoperative and postoperative differential renal function, as measured by nuclear renography. RESULTS: We identified 89 patients, of whom 51 (57%) and 38 (43%) presented with prenatal and postnatal hydronephrosis, respectively. Kidneys in which hydronephrosis was diagnosed prenatally had better average differential renal function than those in which the condition was detected postnatally (45 versus 37%). This difference was even more significant in kidneys with less than 40% initial function (31 versus 21%). Presentation with a palpable mass was associated with worst renal function (mean 23%). Postoperatively renal function did not recover significantly in either group. There was a minimal increase in postoperative differential renal function in the subgroup of patients in whom initial differential renal function was less than 40%, although there was no significant difference in the 2 groups (6.5 versus 4.8%). CONCLUSIONS: The early diagnosis of hydronephrosis provided by prenatal ultrasonography is associated with less obstructive nephropathy. Prolonged followup is necessary for studies of the natural history of hydronephrosis because relevant obstruction manifests clinically years later. Despite successful pyeloplasty function recovery is minimal in kidneys with poor function and hydronephrosis diagnosed prenatally. Our findings do not support previous observations that poor function markedly improves after obstruction is relieved. PMID- 10458426 TI - Elevated bladder urine concentration of transforming growth factor-beta1 correlates with upper urinary tract obstruction in children. AB - PURPOSE: We evaluated urinary transforming growth factor-beta1 (TGF-beta1) concentration in children with upper urinary tract obstruction as a potential tool for supporting the diagnosis of clinically significant obstruction. MATERIALS AND METHODS: Renal pelvic and bladder urine samples were obtained for analysis from 30 patients a median of 5 months old who underwent surgery for obstruction at the ureteropelvic (29) and ureterovesical (1)junctions. Urinary TGF-beta1 concentration was measured using a quantitative sandwich enzyme-linked immunoassay technique. Bladder urine TGF-beta1 in patients with obstruction was compared with that in controls. In addition, we compared renal pelvic and bladder urine TGF-beta1 in patients with obstruction. RESULTS: Mean bladder urine TGF beta1 plus or minus standard error of mean was 4-fold higher in patients with upper tract obstruction than in controls (195 +/- 29 versus 47 +/- 7 pg./mg. creatinine, p <0.001). In the obstructed group mean TGF-beta1 in the renal pelvic urine was 378 +/-86 pg./mg. creatinine, or twice that of the bladder urine (p = 0.02). CONCLUSIONS: Bladder urine TGF-beta1 in patients with upper urinary tract obstruction is significantly elevated compared with that in controls. To our knowledge our study is the first to identify a bladder urinary marker that correlates with upper urinary tract obstruction with greater than 90% sensitivity. Measuring TGF-beta1 in a voided bladder urine sample may provide an objective and noninvasive test for assisting in the diagnosis of upper urinary tract obstruction. PMID- 10458427 TI - Does early detection of ureteropelvic junction obstruction improve surgical outcome in terms of renal function? AB - PURPOSE: Prenatal ultrasound leads to the early detection of hydronephrosis. When followed by the early diagnosis of ureteropelvic junction obstruction surgical treatment, if indicated, is associated with minimal morbidity. We attempt to prove the benefits of this therapeutical approach from the point of view of renal function. MATERIALS AND METHODS: From 1979 to 1997, 452 patients with ureteropelvic junction obstruction underwent dismembered pyeloplasty. Of these patients 113 with comparable data were retrospectively evaluated. Patients were divided into group 1-50 who presented with prenatal hydronephrosis and group 2-63 with neonatal hydronephrosis who were lost to followup and who then presented with symptoms leading to the diagnosis of ureteropelvic junction obstruction. In each group the diagnostic criteria and indications for surgery were identical based on symptomatology, ultrasound and renal scan. Patients with bilateral disease, solitary kidney or vesicoureteral reflux were excluded from study. In groups 1 and 2 median patient age at surgery was 11 months and 5 years, and average postoperative followup was 4.2 and 3.2 years, respectively. RESULTS: In 41 of the 50 group 1 and 60 of the 63 group 2 patients hydronephrosis was severe and the renal pelvis was more than 3 cm. wide. Poor relative renal function (less than 30%) was significantly more pronounced in group 2 than in group 1 (56 of 63 patients or 89% versus 6 of 50 or 12%, p <0.05). In all 113 patients postoperative followup ultrasound and renal scan revealed significant improvement in hydronephrosis and washout curve pattern. There was a significant difference in the 2 groups in regard to renal function improvement postoperatively (66% of group 1 versus 16% of group 2 patients, p <0.05). CONCLUSIONS: We believe that patients who undergo prenatal imaging and are diagnosed early with ureteropelvic junction obstruction should be carefully and meticulously followed. Special efforts should be made to ensure that patients are not lost to followup. After any sign of deterioration develops early surgery is indicated. According to our data operative treatment results in improved renal function. PMID- 10458428 TI - Renal function before and after pyeloplasty: does it improve? AB - PURPOSE: Controversy exists concerning the timing of surgical correction of presumed ureteropelvic junction obstruction. Different opinions on the timing of intervention focus on renal function deterioration with time, poor initial relative function, or obstructive drainage curves and/or drainage time on diuretic renography. We retrospectively determined whether there is any improvement in renal function after pyeloplasty for presumed renal obstruction. MATERIALS AND METHODS: We retrospectively reviewed the charts of patients who underwent pyeloplasty between 1990 and 1997 in whom preoperative and postoperative diuretic renography data were available. Patients were excluded from review when they had bilateral ureteropelvic junction obstruction, a solitary kidney, associated vesicoureteral reflux, or other bladder or ureteral abnormalities, and when preoperative and postoperative renography studies were not available. RESULTS: Complete data were available for review in 79 patients 2 weeks to 18 years old (median age 6 months). Of the patients 73% were male and 73% of the affected kidneys were on the left side. Prenatal hydronephrosis had been diagnosed in 58 patients (73%), of whom 19 (33%) were observed for a variable period before pyeloplasty was performed. In all cases diuretic renography performed at the same institution using a standard protocol revealed a drainage time of 20 minutes or greater preoperatively, while in 58 cases a measurable drainage time was never achieved. As a rule, drainage improved postoperatively (mean and median 25 and 16 minutes, respectively). Open renal biopsy done at pyeloplasty in 54 patients was normal in 29. Preoperatively renal function ranged from 5 to 67% (mean and median 41 and 45, respectively). In all patients the paired t test showed no statistical difference in preoperative and postoperative renal function (p = 0.078, 95% confidence interval -3.451 to 0.185). There was no statistical change in renal function in patients with an abnormal renal biopsy regardless of the severity of renal scarring (p = 0.38) or when renal function was 40% or less (mean preoperative versus postoperative 29.7 versus 28.4%, p = 0.46). The group with greater than 40% function preoperatively had no relevant difference in function before or after surgery (mean 49.7 versus 47.8%, p = 0.065). Prenatally screened patients who were initially observed had a statistically significant difference in renal function before and after pyeloplasty (mean 45.6 versus 43%, p = 0.002). CONCLUSIONS: Renal function did not improve after pyeloplasty regardless of the initial level of relative function. Renal scan revealed that differential function decreased after pyeloplasty in some patients in whom hydronephrosis was detected prenatally and who were initially followed with observation. In our opinion waiting for renal function to decrease before considering pyeloplasty is not warranted, since function does not improve even when obstruction is corrected and drainage time improves. PMID- 10458429 TI - Laparoscopic Anderson-Hynes dismembered pyeloplasty in children. AB - PURPOSE: The feasibility and results of laparoscopic Anderson-Hynes dismembered pyeloplasty in children were evaluated. MATERIALS AND METHODS: All laparoscopic Anderson-Hynes pyeloplasties performed by the author were retrospectively reviewed. A total of 18 children 3 months to 15 years old (mean age 17 months) with proved ureteropelvic junction obstruction underwent laparoscopic Anderson Hynes dismembered pyeloplasty between August 1994 and June 1998. Of the 18 pyeloplasties 15 were performed in children who had not undergone previous upper tract surgery, and 3 had undergone previous upper tract surgery, including laparoscopic pyeloplasty in 2 and emergency percutaneous nephrostomy drainage of pyonephrosis 6 weeks earlier in 1. All operations were performed via a transperitoneal route. RESULTS: Postoperative evaluation is complete in 16 patients and pending in 2. Of the 16 patients 14 (87%) have no demonstrable evidence of obstruction. Two patients with persistent obstruction underwent repeat laparoscopic pyeloplasty. There was no conversion to open surgery. Mean operative time was 89 minutes. In 1 patient trocar hematoma resolved with bed rest. In another case the stent was misplaced with its distal end reaching the lower ureter, and was removed via ureteroscopy 6 weeks postoperatively. There was no other operation related morbidity. CONCLUSIONS: Laparoscopic Anderson-Hynes pyeloplasty represents an attractive alternative to conventional open pyeloplasty. It is technically challenging but with practice it may be completed in the same time as conventional open pyeloplasty. It offers results approaching those of conventional dismembered pyeloplasty. PMID- 10458430 TI - Bladder dysfunction in children with refractory monosymptomatic primary nocturnal enuresis. AB - PURPOSE: We studied bladder dysfunction in children with significant primary nocturnal enuresis refractory to treatment. MATERIALS AND METHODS: We evaluated 33 Chinese boys and 8 girls with a mean age of 10.4 years, who had significant monosymptomatic primary nocturnal enuresis (3 or more wet nights weekly) after desmopressin treatment with or without an enuretic alarm failed. Daytime cystometry, continuous nighttime cystometry and electroencephalography monitoring during sleep, and detailed recording of daytime and nighttime urinary output were performed. RESULTS: We recognized 5 patterns of bladder dysfunction and its association with sleep-arousal status. Pattern 1 was normal daytime urodynamics with significant bladder instability at night with normal volume voiding precipitated by unstable detrusor contractions in 14 boys (34%). Pattern 2 was normal daytime urodynamics with frequent small volume voiding at night, probably representing latent bladder instability, in 4 boys (10%). Pattern 3 involved abnormal daytime urodynamics with small bladder capacity, a discoordinated daytime voiding pattern and marked nighttime bladder instability associated with poor sleep in 6 boys (15%). Pattern 4 was abnormal daytime urodynamics with an obstructive pattern, and marked daytime and nighttime detrusor hypercontractility (mean maximum detrusor pressure 178 cm. water) in 8 boys (20%). Pattern 5 was abnormal daytime urodynamics with a dysfunctional daytime voiding pattern and frequent small volume nighttime voiding in 8 girls and 1 boy (22%). In all patients functional bladder capacity was smaller than expected for age and the majority had no nocturnal polyuria. Despite underlying bladder dysfunction a 4 week course of 400 microg. desmopressin orally at bedtime still produced a significant response with a greater than 50% decrease in the number of wet nights during treatment in 47% of the patients, although enuretic symptoms immediately relapsed on cessation of therapy in all. Notably cystourethroscopy in 7 of the 8 boys with pattern 4 dysfunction revealed bladder trabeculations and abnormal urethral lesions, including congenital obstructive posterior urethral membranes in 4, Moormann's ring in 2 and irregular scarring at the bulbous urethra in 1. CONCLUSIONS: Abnormal bladder function, including small functional capacity, instability during sleep and marked detrusor hypercontractility, was common in our enuretic children in whom treatment failed. More importantly, nocturnal enuresis may be the only presenting symptom and there may be a response to desmopressin with a decreased number of wet nights even in cases of significant underlying bladder dysfunction. These findings may have important implications for our management strategy for monosymptomatic primary nocturnal enuresis. PMID- 10458431 TI - Pelvic floor muscle retraining for pediatric voiding dysfunction using interactive computer games. AB - PURPOSE: We evaluated a new noninvasive outpatient method of pelvic muscle retraining in children using computer game assisted biofeedback. MATERIALS AND METHODS: All patients in whom voiding dysfunction was confirmed by history, uroflowmetry-electromyography and voiding cystourethrography were enrolled in a pelvic floor muscle retraining program. Patients received a pretreatment, mid treatment and posttreatment survey instrument documenting subjective improvement, including the frequency of diurnal enuresis, nocturnal enuresis, constipation and encopresis. Pretreatment and posttreatment simultaneous uroflowmetry surface electrode electromyography was performed and post-void residual urine volume was determined in all patients. RESULTS: A total of 8 boys and 33 girls 5 to 11 years old (mean age 7.2) completed therapy and were available for evaluation. These patients completed 2 to 11 (average 6) hourly treatment sessions. Followup was 3 to 15 months (average 7). At the midterm evaluation improvement in nocturnal enuresis was reported by 57% of the patients, diurnal enuresis by 84%, constipation by 83% and encopresis by 91%. End treatment evaluation revealed improvement in nocturnal enuresis by 90% of patients, diurnal enuresis by 89%, constipation by 100% and encopresis by 100%. Uroflowmetry-electromyography patterns improved in 42% of the patients and post-void residual urine decreased in 57%. Comparison of initial to end recorded millivoltage pelvic floor muscle values demonstrated that 56% of the patients had lower resting tone at the beginning of the session after completing therapy and 78% had improved contracting tone after performing Kegel exercises, as proved by increased microvoltage values. Initial uroflowmetry-electromyography revealed certain categories of cases, including a flattened voiding curve with a hyperactive pelvic floor and low post-void residual urine in 40%, a flattened voiding curve with a hyperactive pelvic floor and high post-void residual urine in 40%, a staccato voiding curve with a hyperactive pelvic floor and low post-void residual urine in 3%, and a staccato voiding curve with a hyperactive pelvic floor and high post-void residual urine in 17%. Of the girls 91% presented with the classic spinning top deformity on voiding cystourethrography. A total of 22 patients presented with a significant history of recurrent urinary tract infections, and infection developed in 3 during treatment and followup. Vesicoureteral reflux in 14 patients resolved during treatment in 3, reimplantation was performed in 1 and 10 are still being observed. CONCLUSIONS: A program of conservative medical management with computer game assisted pelvic floor muscle retraining resulted in significant subjective improvement in continence, constipation and encopresis as well as objective improvement in uroflowmetry-electromyography, post-void residual urine volume and the microvoltage value of pelvic floor muscles in the majority of patients with dysfunctional voiding. PMID- 10458432 TI - alpha-Adrenergic blockade in children with neuropathic and nonneuropathic voiding dysfunction. AB - PURPOSE: Inadequate bladder emptying is a common urinary dysfunction in children. The role of alpha-blockers for managing bladder outlet obstruction remains relatively unexplored in children. Because of the well established impact of alpha-blocker therapy in men, we investigated its use for treating inadequate bladder emptying in the pediatric population. MATERIALS AND METHODS: We treated 17 children 3 to 15 years old with documented poor bladder emptying of various etiologies, including dysfunctional voiding, the Hinman syndrome, the lazy bladder syndrome, posterior urethral valves, myelomeningocele and the prune-belly syndrome, using the alpha-1 adrenergic receptor antagonist, doxazosin. The initial dose of 0.5 to 1.0 mg. nightly was increased according to patient response and as tolerated. Patients were followed weekly to monthly by symptomatic history, and urine flow and/or post-void residual urine volume measurement. Two patients with neurogenic bladder were also followed with cystometrography and leak point pressure determination. RESULTS: Bladder symptomatology and/or emptying improved in 14 patients (82%). Ten patients had decreased post-void residual urine during treatment and in 3 uroflowmetry showed increased maximum flow. Two patients with neuropathic bladder secondary to myelomeningocele had decreased leak point pressure on alpha-blocker therapy and in 2 with a history of posterior urethral valves new onset bilateral hydronephrosis completely resolved. Only 1 patient had mild postural hypotension, which resolved with dose reduction. CONCLUSIONS: Selective alpha-blocker therapy seems to be well tolerated in children and appears effective for improving bladder emptying in various pediatric voiding disorders at short-term followup. Long-term followup and further investigation are warranted to validate the potential role of alpha-blocker therapy in pediatric urinary dysfunction. PMID- 10458433 TI - Improved bladder function after prophylactic treatment of the high risk neurogenic bladder in newborns with myelomentingocele. AB - PURPOSE: High pressure dyssynergic voiding may result in irreversible damage to the urinary tract. Prophylactic therapy in the form of clean intermittent catheterization and anticholinergic medication may significantly decrease the incidence of upper urinary tract deterioration. Whether prophylactic therapy in the high risk bladder may also lead to improved long-term bladder dynamics prompted us to study the effect of early versus late treatment of bladder hypertonicity and detrusor-sphincter dyssynergia on the ultimate need for bladder augmentation. MATERIALS AND METHODS: We retrospectively reviewed urological outcomes in patients with myelodysplasia who were at risk for urological deterioration within year 1 of life based on bladder sphincter dyssynergia and/or high filling or voiding pressure. We recorded the dates when high risk voiding dynamics were initially observed, and when intermittent catheterization and anticholinergic therapy were initiated. Patients in whom treatment began at the time a high risk profile was noted (prophylactic group seen between 1985 and 1990) were compared to controls with the same high risk voiding parameters who did not receive early therapy (observation group seen between 1978 and 1984 with therapy instituted 1 year or longer after high risk was noted). The number of augmentations performed in each group was indexed to the total number of years of followup in the 2 populations, respectively. Patients with less than 2 years of followup were excluded from further analysis. RESULTS: Of the 45 patients at risk clean intermittent catheterization and anticholinergic medication were immediately initiated in 18, while 27 were treated expectantly. Patients in the observation group were followed an average of 4.1 years (range 1.1 to 14) before clean intermittent catheterization and anticholinergic medication were started. Of the 27 children treated expectantly 11 (41%) required augmentation, whereas only 3 of the 18 (17%) treated prophylactically required enterocystoplasty. When the number of augmentations was indexed to total years of followup in each of the 2 groups (296 versus 156 years) patients in the expectant group were nearly twice as likely to require augmentation. CONCLUSIONS: Identification and early proactive treatment of the high pressure, dyssynergic lower urinary tract significantly decreases the need for bladder augmentation as children with neurogenic bladder secondary to myelomeningocele mature. PMID- 10458434 TI - Long-term outcome based on the initial surgical approach to ureterocele. AB - PURPOSE: The management of extravesical ureterocele is controversial. Heminephrectomy and recently recommended primary incision or puncture have high reoperation rates. We reviewed and compared the long-term results of these procedures with those of primary lower tract reconstruction for ureterocele. MATERIALS AND METHODS: We reviewed the records of 106 children with ureterocele treated between 1979 and 1997. Followup was available in 99 patients, including 72 with extravesical and 27 with intravesical ureterocele. Patients with extravesical ureterocele were divided based on initial management into group 1-13 who underwent transurethral incision or puncture, group 2-41 who underwent an upper tract approach, including partial or complete nephrectomy with partial ureterectomy or ureteroureterostomy and group 3-18 who underwent complete reconstruction, including ureterocelectomy and ureteral reconstruction with or without upper tract surgery. RESULTS: Overall the reoperation rate in patients with intravesical ureterocele was 22% and 23% in those treated with initial endoscopic incision or puncture. In patients with extravesical ureterocele the reoperation rate was 100, 41 and 0% in groups 1 to 3, respectively. Differences in followup (overall mean 6 years) and the incidence of preoperative reflux in the 3 groups were not statistically significant. In group 2, the reoperation rate in patients with versus without preoperative reflux was 57 versus 20% (p = 0.08). Of the 25 prenatally diagnosed patients urinary tract infection developed preoperatively in 3 (12%) at ages 2, 3, and 6 months, respectively. Mean age at the time of the initial operation in all prenatally diagnosed patients was 3.1 months (range 5 days to 11 months). CONCLUSIONS: Complete reconstruction appears to be safe and highly effective even in infancy for treating extravesical ureterocele. Although the primary upper tract approach is associated with a significantly higher reoperation rate, it is a favorable alternative in patients with no preoperative reflux. However, while transurethral decompression is effective in the majority of patients with intravesical ureterocele, it is not definitive therapy for extravesical ureterocele and it should have a limited role in initial management. PMID- 10458435 TI - Long-term outcome of transurethral puncture of ectopic ureteroceles: initial success and late problems. AB - PURPOSE: We studied the long-term outcome of transurethral puncture of ectopic ureteroceles specifically associated with duplex systems. MATERIALS AND METHODS: We retrospectively reviewed the records of patients who underwent transurethral puncture of an ectopic ureterocele. Study exclusion criteria were orthotopic, bilateral and prolapsing ureteroceles. RESULTS: We identified 19 girls and 2 boys, of whom 11 presented with prenatal hydronephrosis and 10 presented with urinary tract infection. Mean age at puncture was 5 months (range 0.5 to 60). Preoperatively voiding cystourethrography revealed no reflux in 7 patients, isolated ipsilateral lower pole reflux in 8, and bilateral and/or contralateral reflux in 6. Postoperatively studies initially showed no reflux in 8 cases but in 4 of the 8 reflux recurred up to 4 years after puncture. In 10 patients (48%) reflux developed into the ureterocele and upper pole segment. Repeat puncture was required 1 to 13 months after the initial procedure in 4 patients for persistent or recurrent upper pole hydroureteronephrosis. Subsequent open surgery was required in 15 of the 21 cases (71%), including ureterocele excision with ureteral reimplantation in 14. Of the children 10 and 4 underwent open surgery for recurrent urinary tract infection and progressive reflux, respectively, while 1 underwent ureteroureterostomy for progressive upper pole reflux. No patient underwent upper pole nephrectomy. Of the remaining 6 patients 4 have low grade reflux. CONCLUSIONS: Transurethral puncture of ectopic ureteroceles provides effective short-term correction of upper pole obstruction but it is not definitive therapy in the majority of cases. Most children still require open surgery. In patients without reflux after the puncture procedure new onset, recurrent or progressive reflux may later develop with extended followup. Repeat puncture may be required to ensure adequate decompression in a minority of cases, as in the 20% in our series. PMID- 10458436 TI - Associated urological anomalies in children with unilateral renal agenesis. AB - PURPOSE: Unilateral renal agenesis has been noted in 1:1,000 autopsies. Recently an increased incidence of vesicoureteral reflux was reported in patients with a solitary kidney. We determined the incidence of associated renal abnormalities in children with unilateral renal agenesis. MATERIALS AND METHODS: We retrospectively reviewed 46 consecutive cases of unilateral renal agenesis diagnosed at our hospital between January 1985 and February 1998. Patient age at diagnosis ranged from newborn to 12.5 years (mean 2.8 years). There were 24 boys and 22 girls. The left kidney was absent in 27 patients and the right kidney was absent in the remaining 19. A total of 24 patients were evaluated for urinary tract infection and in the other 22 a solitary kidney was found during examination for congenital malformations, enuresis or abdominal pain. The diagnosis was made in all patients by abdominal ultrasound and confirmed by excretory urography, or diethylenetriaminepentaacetic acid or dimercapto-succinic acid scan. A voiding cystourethrogram was performed in 40 patients (87%). RESULTS: Associated urological anomalies were present in 22 of the 46 patients (48%) with unilateral renal agenesis, including primary vesicoureteral reflux in 13 (28%), ureterovesical junction obstruction in 5 (11%), ureteropelvic junction obstruction in 3 (7%), and ureterovesical and ureteropelvic junction obstruction in 1 (2%). Of the 22 patients 14 (64%) underwent surgical intervention. CONCLUSIONS: Nearly half of the patients with unilateral renal agenesis had associated urological anomalies. Vesicoureteral reflux was the most common associated anomaly and it was usually of high grade. Early recognition and treatment of urological anomalies in a patient with a solitary kidney are imperative to decrease the long-term risk of renal damage. PMID- 10458437 TI - Contralateral compensatory kidney growth in rats with partial unilateral ureteral obstruction monitored by magnetic resonance imaging. AB - PURPOSE: We studied dynamic changes in total volume and renal vein blood flow in the kidneys contralateral to partial ureteral obstruction induced in newborn rats. MATERIALS AND METHODS: Using magnetic resonance imaging we investigated changes in total kidney volume and renal vein blood flow in 20 rats with mild and 18 with severe partial ureteral obstruction that was induced on the left side 2 days after birth. A total of 15 sham operated control rats were also studied. Total kidney volume and renal vein blood flow were monitored sequentially every 2 to 6 weeks for a total of 24 weeks. Renal parenchymal volume measured in vivo by magnetic resonance imaging was compared with that measured in vitro at week 24. RESULTS: Total volume and renal vein blood flow increased significantly in contralateral nonobstructed kidneys from week 14 and thereafter in rats with severe partial unilateral ureteral obstruction. At week 24 volume had increased by 22% and blood flow had increased by 25%. Volume and flow did not increase significantly in contralateral nonobstructed kidneys in rats with mild partial unilateral ureteral obstruction (p = 0.09). Before changes occurred in volume and blood flow in the contralateral nonobstructed kidneys, renal vein blood flow had decreased significantly from week 8 and thereafter in severely obstructed kidneys. In rats with mild partial unilateral ureteral obstruction renal vein blood flow decreased significantly in obstructed kidneys from week 18 and thereafter. Good correlation was noted between MRI in vivo and in vitro kidney volume measurements (r = 0.972, p <0.001). CONCLUSIONS: A significant increase in total volume and renal vein blood flow in contralateral nonobstructed kidneys did not develop immediately after the onset of detectable functional deterioration in partially obstructed kidneys. Therefore, caution should be used when incorporating compensatory growth into surgical decision making. PMID- 10458438 TI - Experimental fetal vesicoureteral reflux induces renal tubular and glomerular damage, and is associated with persistent bladder instability. AB - PURPOSE: We assessed renal function and urodynamic status in animals with experimental congenital vesicoureteral reflux. MATERIALS AND METHODS: Vesicoureteral reflux was surgically induced in male sheep fetuses at 95 days of gestation. After birth the animals were maintained on antibiotic prophylaxis. At ages 1 week and 6 months reflux was assessed by fluoroscopic voiding cystography. Cystometrography was performed with the animals awake. Serum creatinine, inulin clearance and the excretion of urinary N-acetyl-beta-D-glucosaminidase were measured at ages 1 week, 1 month and 6 months by surveillance urine cultures. Urinary concentrating capacity was assessed by desmopressin testing at ages 1 and 6 months. RESULTS: Nine animals (18 renal units) were born after the induction of reflux. There was no reflux in 2 renal units, while reflux was mild in 2, moderate in 5 and severe in 9. In the 6 animals available for followup at age 6 months only severe reflux persisted. Reflux resolution was associated with normalization of bladder urodynamics. Surveillance urine cultures were negative until age 6 months, when infection developed in 3 of the 6 lambs. In all animals serum creatinine was normal during followup. Glomerular filtration rate in the lambs with reflux was no different from normal at age 1 week but it was significantly less than normal independent of infection at age 6 months (2.7 versus 3.9 ml./kg. per minute, p = 0.002). As an indicator of renal tubular injury the ratio of N-acetyl-beta-D-glucosaminidase-to-creatinine remained significantly higher in animals with reflux than in normal animals from ages 1 week to 6 months (51.0 versus 10.2 IU/mg., p = 0.03). Maximal concentrating ability after desmopressin testing was already less than normal by age 1 month with a maximal increase of 98 versus 435 mOsm./l. in lambs with reflux versus normal lambs (p <0.0001). It was further impaired by age 6 months. Urodynamic evaluation of the animals with reflux revealed decreased bladder compliance at age 1 week with normal voiding pressure. In addition, in those with reflux there was a more pronounced immature voiding pattern with multiple phasic contractions due to sphincteric activity as well as a post-void bladder contraction. CONCLUSIONS: Our model of fetal vesicoureteral reflux induces alterations in renal function that are consistent with clinical observations and marked by altered tubular function but a relatively mild decrease in glomerular filtration. Bladder dynamics are altered, consistent with observations in human neonates with high grade reflux and bladder instability. Whether this represents cause or effect remains unclear. Our model permits focused study of the interaction of these factors in neonatal reflux and may allow the application of more specific therapies, particularly those directed toward mechanisms of renal and bladder dysfunction. PMID- 10458439 TI - New concepts of histological changes in experimental augmentation cystoplasty: insights into the development of neoplastic transformation at the enterovesical and gastrovesical anastomosis. AB - PURPOSE: To our knowledge the pathogenesis of malignancy associated with ileal cystoplasty, ureterosigmoidostomy and ileal conduits is currently unknown. To gain further insights into the mechanism of neoplastic transformation we studied histological changes in a canine augmentation cystoplasty model. MATERIALS AND METHODS: Enterocystoplasty and gastrocystoplasty were performed using a 5 to 7 cm. patch of ileum in 8 dogs and gastric antrum in 6. Specimens were harvested 4 months postoperatively. Representative 3 microm sections of the enterovesical and gastrovesical junctions were stained with hematoxylin and eosin. Uroplakin expression was assessed using an indirect peroxidase method subjected to double staining with alcian blue and periodic acid-Schiffreagent. RESULTS: The bladder portion of the augmentation cystoplasty had 3 to 4 stratified cell layers covered with a distinctive umbrella cell layer. Strong uroplakin staining was visible in all cell layers except the basal layer. At the enterovesical and gastrovesical junctions 6 to 10 layers of hyperplastic, urothelial appearing cells covered the glandular epithelium of the ileal and gastric segments. These cells expressed uroplakins. At this junction zone there was a marked decrease of underlying enteric glands, which had atrophied in proportion to the degree of urothelial hyperplasia. Double staining of uroplakin stained sections with alcian blue and periodic acid-Schiff reagent revealed mucosubstances in hyperplastic urothelial cells covering the enteral segments, indicating that the cells co-expressed uroplakins and mucins. CONCLUSIONS: Histological changes in this experimental canine model of augmentation cystoplasty indicated that the overgrowth of hyperplastic transitional epithelium develops at the enterovesical and gastrovesical junctions. These cells express not only uroplakins, but also mucosubstances. Our results suggest that the migrated hyperplastic urothelial cells have undergone changes characteristic of the enteric and gastric epithelium, which may have important implications in the pathogenesis of malignancy in bladder augmentations. PMID- 10458440 TI - Understanding bladder regeneration: smooth muscle ontogeny. AB - PURPOSE: We determined the origin of smooth muscle cells in acellular bladder matrix grafts. MATERIALS AND METHODS: A total of 15 female Sprague-Dawley rats underwent partial cystectomy and grafting with an acellular matrix derived from rat bladder. The grafts were examined 1, 2, 3 and 4 weeks after grafting by immunohistochemical studies for smooth muscle markers and by transmission electron microscopy for smooth muscle morphology. Bladder matrix and bladder epithelium recombinants were created and grafted subcutaneously and under the renal capsule in nude mice. Recombinants were examined 1, 2, 3 and 4 weeks postoperatively by immunohistochemical studies for bladder epithelium and bladder smooth muscle. RESULTS: Smooth muscle ingrowth into acellular matrix was initially seen at 2 weeks. The immunohistochemical and electron microscopic characteristics of the cells were similar to those of fetal smooth muscle 2 weeks and newborn smooth muscle 4 weeks after grafting. Matrix epithelium recombinants displayed mature bladder epithelium with 3 to 7 layers but they did not support the ingrowth of smooth muscle cells. CONCLUSIONS: Mature bladder smooth muscle cells undergo dedifferentiation, migration and redifferentiation to repopulate an acellular matrix graft. It is unlikely that adult fibroblasts from the surrounding tissue are induced by epithelium and matrix to form smooth muscle. The contractile behavior of bladder substitute materials likely reflects the properties of the host bladder. PMID- 10458441 TI - Reconstitution of human corporal smooth muscle and endothelial cells in vivo. AB - PURPOSE: The availability of autologous erectile tissue composed of corporal smooth muscle and endothelial cells would be beneficial in patients undergoing penile reconstruction. We previously showed that cultured cavernous cells seeded on polymer scaffolds form corporal muscle when implanted in vivo. However, to reconstruct corporal tissue endothelial and corporal muscle cells are necessary. In this study we investigated the possibility of developing tissue composed of corporal cells in vivo by combining smooth muscle and endothelial cells. MATERIALS AND METHODS: Human corporal smooth muscle and endothelial cells were seeded on biodegradable polyglycolic acid polymer scaffolds at concentrations of 20 x 10(6) and 10 x 10(6) cells per cm3, respectively. A total of 60 polymer scaffolds seeded with cells and 20 control polymers without cells were implanted in the subcutaneous space of 20 athymic mice. Mice were sacrificed 1, 3, 5, 7, 14, 21, 28 and 42 days, respectively, after implantation. Immunocytochemical and histochemical analyses were performed with antifactor VIII, antipancytokeratins and anti-alpha actin antibodies. RESULTS: Histologically the retrieved polymers seeded with corporal smooth muscle and endothelial cells showed the formation of multilayered smooth muscle strips adjacent to endothelial cells 7 days after implantation. Increased organization of the smooth muscle tissue and accumulation of endothelium lining the luminal structures were evident by 14 days. A well organized tissue construct was noted 28 and 42 days after implantation. There was no evidence of tissue formation in controls. Immunocytochemical analysis using antifactor VIII to identify native vasculature only and antipancytokeratins to identify ECV 304 endothelial cells only distinguished the origin of the vascular structures in each construct. Anti-alpha-actin confirmed the smooth muscle phenotype. CONCLUSIONS: Human corporal smooth muscle and endothelial cells seeded on biodegradable polymer scaffolds formed vascularized corpus cavernosum muscle when implanted in vivo. To our knowledge this is the first demonstration in tissue engineering in which capillary formation was facilitated by the addition of endothelial cells in composite tissue in vivo. PMID- 10458442 TI - Oxybutynin chloride inhibits proliferation and suppresses gene expression in bladder smooth muscle cells. AB - PURPOSE: We test the hypothesis that oxybutynin chloride inhibits bladder smooth muscle cell proliferation. MATERIALS AND METHODS: Cultured rat bladder smooth muscle cells were grown in Medium 199 supplemented with 10% fetal bovine serum in the presence of 0, 1, 10 and 100 microM. oxybutynin. Cell proliferation was assessed by counting cell numbers 48 and 96 hours after plating. To investigate the role of oxybutynin in bladder smooth muscle cell proliferation after mechanical stretch, cells were grown on silicone elastomer bottomed culture plates and subjected to cyclical stretch-relaxation for 48 hours in the presence of 10 microM. oxybutynin. Deoxyribonucleic acid synthesis was assessed by tritiated thymidine incorporation assay. To examine the effect of oxybutynin on stretch activated gene expression, bladder smooth muscle cells were subjected to stretch-relaxation for 2 hours with and without 10 microM. oxybutynin, and relative c-jun messenger (m) ribonucleic acid (RNA) levels were assessed by semiquantitative reverse transcriptase-polymerase chain reaction with normalization to glyceraldehyde-3-phosphate dehydrogenase mRNA levels. RESULTS: The serum stimulated increase in bladder smooth muscle cell growth was inhibited by oxybutynin in a dose dependent manner. In bladder smooth muscle cells there was a 4.7-fold increase in deoxyribonucleic acid synthesis after mechanical stretch, which decreased by 40% (p <0.01) when cells were stretched in the presence of oxybutynin. Stretch stimulated significant increase in c-jun mNRA levels, which was significantly decreased by oxybutynin. CONCLUSIONS: Oxybutynin chloride inhibits bladder smooth muscle cell proliferation induced by serum and mechanical stretch. A potential mechanism by which oxybutynin inhibits proliferation may be the down regulation of growth promoting genes, such as c jun. We speculate that oxybutynin may be useful for preventing permanent hypertrophic bladder changes in addition to decreasing intravesical pressure. PMID- 10458443 TI - Direct in vivo gene transfer to urological organs. AB - PURPOSE: Patients with urological disorders may benefit from gene based therapy. We investigated the feasibility of delivering exogenous genes into urological tissues in vivo using direct in vivo electrotransfection. MATERIALS AND METHODS: Gene transfer to rat kidneys, testes and bladders was accomplished via direct local injection of pGL3/luciferase and beta-galactosidase reporter gene constructs, followed by an electrical pulse ranging from 55 to 115 msec at 100 V. Direct injection of deoxyribonucleic acid without an electrical pulse served as the control. The transfected and nontransfected organs were retrieved and analyzed by luciferase activity assay, histochemical and immunocytochemical staining for beta-galactosidase, and reverse transcription polymerase chain reaction with primers specific for beta-galactosidase messenger ribonucleic acid. RESULTS: There was significant luciferase activity 1, 3 and 5 days after direct in vivo electrotransfection in kidneys and testes, and after 3, 5, 7 and 10 days in bladders. Positive beta-galactosidase enzyme activity and beta-galactosidase immunoreactivity were observed in the transfected renal tubular cells, testicular interstitial and germ cells, and uroepithelial bladder layer. Reverse transcription-polymerase chain reaction products of the transfected organs were noted, indicating the successful transcription of messenger ribonucleic acid. CONCLUSIONS: This study demonstrates that direct in vivo electrotransfection is a feasible method of transient gene delivery into intact urological organs. Its apparent safety and relative simplicity suggest that direct in vivo electrotransfection may be useful clinically. PMID- 10458444 TI - Autologous engineered cartilage rods for penile reconstruction. AB - PURPOSE: Conditions such as inadequate and ambiguous genitalia that are caused by rudimentary penis, severe hypospadias or traumatic injury require surgical intervention. Although silicone penile prostheses are an accepted treatment modality, biocompatibility issues may be a problem in select cases. We previously demonstrated that rods composed of cartilage could be created using chondrocytes seeded on biodegradable polymer scaffolds. We showed that the cartilage rods engineered ex situ were readily elastic and withstood high degrees of pressure. We investigated the feasibility of applying the engineered cartilage rods in situ in an animal model. MATERIALS AND METHODS: Autologous chondrocytes harvested from rabbit ears were grown and expanded in culture. Cells were seeded onto biodegradable poly-L-lactic acid coated polyglycolic acid polymer rods at a concentration of 50 x 10(6) chondrocytes per cm3. A total of 18 chondrocyte polymer scaffolds were implanted into the corporal spaces in 10 rabbits. As controls, 1 corpus in each of 2 rabbits was not implanted. The animals were sacrificed 1, 2, 3 or 6 months after implantation. Histological analysis was performed using hematoxylin and eosin, aldehyde fuschin-alcian blue and toluidine blue staining. RESULTS: All animals tolerated the implants for the duration of the study without any complications. Gross examination after retrieval at 1 month showed well formed, milky white cartilage structures within the corpora. All polymers were fully degraded by 2 months. There was no evidence of erosion or infection at any of the implant sites. Histological analysis using alcian blue and toluidine blue staining revealed mature and well formed chondrocytes in the retrieved implants. CONCLUSIONS: Autologous chondrocytes seeded on preformed biodegradable polymer structures form cartilage structures within the rabbit corpus cavernosum. This technology appears to be useful for creating autologous penile prostheses. PMID- 10458445 TI - Gastrocystoplasty: long-term complications in 22 patients. AB - PURPOSE: Gastrocystoplasty has been performed as an alternative to enterocystoplasty to increase bladder capacity and/or compliance while avoiding the complications associated with the use of bowel segments. Gastrocystoplasty is not without metabolic and physiological complications, such as the dysuria hematuria syndrome and hypochloremic metabolic alkalosis. Currently to our knowledge there is limited long-term followup of gastrocystoplasty, which prompted us to review our experience with gastrocystoplasty and compare our findings with those of others. MATERIALS AND METHODS: We retrospectively reviewed for complications the records of 12 boys and 10 girls 8 to 24 years old who underwent gastrocystoplasty. Followup ranged from 48 to 96 months. The diagnosis included neurogenic bladder in 12 cases, posterior urethral valves in 6, bladder exstrophy in 3 and pelvic tumor in 1. All patients underwent preoperative evaluation of serum electrolytes, blood urea nitrogen and creatinine as well as a radionuclide renal scan. Urodynamics were done preoperatively in all patients and postoperatively for complications. A gastric wedge with the pedicle based on the right gastroepiploic artery was removed, leaving the lesser curvature and vagus nerve intact. This technique was used in 21 of the 22 cases, including 1 case after initial surgery elsewhere. Ureteroneocystotomy, Mitrofanoff appendicovesicostomy and bladder neck reconstruction were performed as indicated. RESULTS: There was 1 early complication (postoperative bleeding) and the remainder were late complications, including vesicoureteral reflux in 4 cases, Mitrofanoff valve stenosis in 3, the hematuria-dysuria syndrome, renal calculi, decreased bladder capacity with incontinence and metabolic alkalosis in 2 each, and ureterovesical stricture in 1. The late complication rate in our series was 36%. CONCLUSIONS: Our long-term results differ from those of others in the number of late complications (36 versus 21.8%). In addition, 50% of our patients with complications had multiple complications. These findings may be due to a longer followup. Nevertheless, our data cast serious doubt on the long-term advantages of using stomach for bladder augmentation. PMID- 10458446 TI - The gastrointestinal composite urinary reservoir in patients with myelomeningocele and exstrophy: long-term metabolic followup. AB - PURPOSE: We investigated the long-term metabolic effects of gastrointestinal composite urinary reservoirs in patients with myelomeningocele or exstrophy. MATERIALS AND METHODS: Seven patients with myelomeningocele or exstrophy who required complex urinary reconstruction in the setting of metabolic acidosis or the short bowel syndrome underwent construction of a gastrointestinal composite reservoir, including a staged and a single procedure in 3 and 4, respectively. Preoperatively and postoperatively serum electrolytes were measured, and urinalysis and urine cultures were performed in all patients. In 5 patients serum pH was compared preoperatively and postoperatively, and in all serum gastrin was measured postoperatively. RESULTS: At an average followup of 62 months (range 52 to 87) serum chloride and bicarbonate significantly normalized (p <0.05) in all 7 patients with bladder exstrophy or myelomeningocele. Serum pH also significantly normalized (p <0.05) in 5 patients at long-term followup. Serum gastrin and creatinine were normal and urinary pH fluctuated insignificantly throughout followup. None of the patients had urolithiasis or symptoms of the hematuria dysuria syndrome. Periodic symptomatic urinary tract infections developed but none required chronic antibiotic therapy. CONCLUSIONS: Gastrointestinal composite urinary reservoirs appear to be beneficial for patients with myelomeningocele or exstrophy who have preexisting metabolic acidosis or the short bowel syndrome. Serum electrolyte neutrality is achieved during long-term followup. No patient had the hematuria-dysuria syndrome or urolithiasis. PMID- 10458448 TI - Bladder augmentation with urothelial preservation. AB - PURPOSE: Bladder reconstruction is performed because the characteristic properties of a healthy bladder are no longer present. The bladder manifests poor capacity, poor compliance and potential or actual changes in the upper tracts that may lead to damage. Augmentation procedures provide a means to improve capacity and compliance, and they may be performed with catheterizable channels to facilitate bladder emptying. Natural tissues or synthetic materials may be used but urothelial preservation is desirable. Demucosalized augmentation with a gastric flap and auto-augmentation with peritoneum are techniques that have been used in the last 5 years. MATERIALS AND METHODS: We retrospectively evaluated the records of 13 patients who underwent demucosalized augmentation with a gastric flap and 8 who underwent augmentation with peritoneum from 1992 to 1995. Average age of the 11 girls and 10 boys was 8 years (range 6 to 12). The diagnosis was myelomeningocele in 15 patients, exstrophy in 2, and the VATER association, posterior urethral valves, spinal cord injury and nonneurogenic neurogenic bladder in 1 each. Concurrent procedures included appendicovesicostomy creation, a fascial sling or wrap and ureteroneocystostomy. RESULTS: Mean followup is 50 months for patients who underwent demucosalized augmentation with a gastric flap and 47 for those who underwent augmentation with peritoneum. Outcome was defined as good-dry for 4 hours, catheterization without difficulty and a stable upper tract; poor-a secondary procedure (augmentation) required because the initial procedure did not improve bladder capacity, compliance, continence or the degree ofhydronephrosis, and fair-dry for less than 4 hours, some problems with incontinence, or compliance 10 ml/cm water or less. Of the patients who underwent demucosalized augmentation with a gastric flap the outcome was good in 5, fair in 4 and poor in 4 who required repeat augmentation. Of the 8 patients who underwent auto-augmentation with peritoneum the outcome was good in 5 and poor in 2, and 1 was lost to followup. CONCLUSIONS: Augmentation with urothelial preservation may result in a good capacity, compliant bladder in certain patients but a poorly compliant, small capacity bladder in others. Our overall results underscore the lack of understanding of the properties and characteristics of these bladders and of stromal-epithelial interaction that occurs after augmentation. Such an understanding is critical before this procedure can be recommended routinely. PMID- 10458447 TI - Clinical and urodynamic evaluation after ureterocystoplasty with different amounts of tissue. AB - PURPOSE: Ureter is one of the best tissues for bladder augmentation. The amount of ureteral segment available is extremely variable among patients. We compared results in patients who underwent ureterocystoplasty with 2 ureters, 1 complete ureter or a distal segment only after transureteroureterostomy. MATERIALS AND METHODS: During a 6-year period we performed 32 ureterocystoplasties at 2 pediatric centers in Argentina (16) and Chile (16). Median patient age at surgery was 9 years (range 4 months to 20 years). Clinical presentation included urinary infection, hydronephrosis, incontinence and undiversion. The diagnosis was neurogenic bladder in 20 cases, infravesical obstruction in 7, massive reflux in 3 and ureterocele in 2. All patients had poor bladder compliance and vesicoureteral reflux. We used different options to augment the bladder, including 2 ureters in 5 patients, bilateral nephrectomy in 3, a complete duplex system in 1 and a bilateral partial ureter in 1 (group 1); a complete ureter in 14 (group 2), and a distal segment of ureter with transureteroureterostomy in 13 (group 3). When transureteroureterostomy was performed, a suprapubic tube remained indwelling for 2 weeks and a Double-J stent was placed for 1 month. Median followup was 16 months (range 4 months to 6 years). Clinical and radiological evaluations, including ultrasound, cystography, urodynamics, renal scan and renal function measurement, were done 4 months postoperatively and twice yearly thereafter as needed. RESULTS: We noted no significant difference in bladder capacity when 1 or 2 ureters were used. Median increase in bladder capacity in groups 1 and 2 was 375% (range 80 to 800). All patients who received a complete segment of ureter had clinical improvement, decreased hydronephrosis and resolution of reflux with improved bladder compliance. When a partial segment of ureter was used median capacity increased 230% (range 40 to 400) with clinical improvement in 12 patients (92.3%). Compliance improved, which led to longer intervals between clean intermittent catheterizations. No patient has needed repeat augmentation to date. CONCLUSIONS: There is a difference in median increased bladder capacity when a segment of distal ureter is used to augment the bladder versus 1 or 2 whole ureters. However, the use of distal ureter still represents a safe alternative for augmenting the bladder and simultaneously resolving massive reflux. Ureterocystoplasty is an excellent choice for increasing bladder capacity and improving bladder compliance despite the different amounts of tissue available. PMID- 10458449 TI - Treatment of the Mayer-Rokitansky-Kuster-Hauser syndrome in Bangladesh: results of 10 total vaginal replacements with sigmoid colon at a missionary hospital. AB - PURPOSE: We report our experience with treating the Mayer-Rokitansky-Kuster Hauser syndrome in a developing country. The operations were performed by a pediatric surgical team at a missionary hospital in Khulna, Bangladesh. MATERIALS AND METHODS: From 1995 to 1998, 10 young women with the Mayer-Rokitansky-Kuster Hauser syndrome underwent complete vaginal replacement. In 4 patients the abnormality was discovered after marriage at the initial sexual approach and, thus, the husband abandoned 3. In 4 of the 10 cases the diagnosis was suspected because of absent menstruation. A physician made the diagnosis in only 2 cases. Preoperatively abdominal ultrasound in 3 patients showed a hypoplastic uterus in all and a right solitary pelvic kidney in 1. In all 10 women a neovagina was created using a 14 cm. segment of sigmoid colon. Two weeks postoperatively patients were taught to dilate and irrigate the neovagina. RESULTS: A minimum of 1 year of followup is available in 7 of the 10 patients. The vagina had a good appearing introitus. Mucous production significantly decreased 3 to 4 months after the operation. Two patients already had an active sexual life. The remaining 3 patients underwent surgery during the last mission and they had no complications 6 months postoperatively. CONCLUSIONS: Our experience shows the feasibility of treating patients with a severe abnormality in a hospital with basic facilities in one of the poorest countries in the world. PMID- 10458450 TI - 5alpha-reductase type 2 mutations are present in some boys with isolated hypospadias. AB - PURPOSE: We determined whether 5a-reductase type 2 mutations are present in boys with isolated hypospadias. MATERIALS AND METHODS: Penile skin tissues obtained at surgery during hypospadias repair were examined for 5alpha-reductase type 2 mutations by single strand conformational polymorphism and deoxyribonucleic acid sequence analysis. Clinical data, including family history of hypospadias and preoperative position of the urethral meatus, were correlated with the genetic findings. RESULTS: Of the 81 specimens examined 7 (8.6%) involved a mutation in at least 1, 5alpha-reductase type 2 gene, while 2 patients had mutations in both alleles. The mutations identified were A49T, L113V and H231R. The A49T mutation in 5 patients was the most common (71%) and it was generally present in less severe forms of hypospadias. To our knowledge neither the A49T nor the L113V mutation has been previously reported in association with 5alpha-reductase type 2 deficiency and to date they have only been identified in cases of isolated hypospadias. Family history was negative in the 7 patients with 5a-reductase type 2 mutations but positive in 5 without mutations. CONCLUSIONS: Some boys with isolated hypospadias have a mutation in at least 1 gene for 5alpha-reductase type 2. This finding suggests that a partial deficiency of 5alpha-reductase activity and inadequate levels of dihydrotestosterone in the fetal urethra may be sufficient to cause the phenotype of hypospadias without other clinical features of 5alpha-reductase deficiency. Family history may not be reliable for determining which boys with hypospadias are likely to have such mutations. PMID- 10458451 TI - Tunica vaginalis onlay urethroplasty as a salvage repair. AB - PURPOSE: We review our experience with tunica vaginalis onlay urethroplasty for urethral repair after multiple previous hypospadias repairs. MATERIALS AND METHODS: From January 1992 through December 1997 we repaired the urethra in 10 boys and 1 man using a tunica vaginalis onlay flap. The tunica vaginalis was placed as a patch on the urethra that was proximal to the glans or brought out to the urethral meatus. RESULTS: No intraoperative complications or difficulties were associated with harvesting the tunica vaginalis. Of the 4 children and 1 adult in whom the onlay flap was brought out to the urethral meatus meatal stenosis developed in 3 (60%), while 2 void without problems. In 6 children the onlay flap was placed proximal to the glans and in 1 who was lost to followup 2 discontinuous patches were placed. In 3 of the 5 remaining boys (60%) stricture developed, while 2 void without problems. CONCLUSIONS: Tunica vaginalis onlay urethroplasty results in a substantial complication rate. We did not confirm any advantage of its use. PMID- 10458452 TI - A novel inert collagen matrix for hypospadias repair. AB - PURPOSE: In select patients with hypospadias in whom genital skin is insufficient alternative tissues are needed for urethral reconstruction. Although skin and mucosal grafts may be used, they may increase hospitalization and morbidity. We explored the feasibility of using a bladder submucosal, collagen based inert matrix as a free graft substitute for urethral repair. MATERIALS AND METHODS: Four patients with a history of hypospadias underwent repeat hypospadias repair using a collagen based inert matrix for urethral reconstruction. The inert collagen matrix was trimmed to size as needed for each patient. The neourethra was created by anastomosing the matrix in an onlay fashion to the urethral plate with continuous 6-zero polyglactin sutures. The created neourethra size ranged from 5 to 15 cm. RESULTS: After a 22-month followup 3 of the 4 patients had a successful outcome in regard to cosmesis and function. One patient in whom a 15 cm. neourethra was created had a subglanular fistula. CONCLUSIONS: The use of a collagen inert matrix appears to be beneficial in patients who have undergone previous hypospadias repair and who may lack sufficient genital skin for reconstruction. PMID- 10458453 TI - Chordee correction by corporal rotation: the split and roll technique. AB - PURPOSE: The optimal approach to correcting ventral chordee associated with severe hypospadias is controversial. Dorsal plication tends to shorten the phallus and ventrally positioned grafts often mandate a staged procedure. An alternative approach is presented using corporal rotation to correct ventral chordee associated with hypospadias. MATERIALS AND METHODS: In 6 boys with severe hypospadias the urethral plate was divided and the septum between the corpora cavernosa was partially split with a ventral midline incision. This incision facilitates corporal rotation. Access to the dorsal aspect of the corpora cavernosa was achieved by dissecting Buck's fascia with its encased neurovascular bundles, so that the bundles in the area of chordee were completely elevated and preserved. Using artificial erection as a guide nonabsorbable sutures were placed in the area of maximal curvature from the dorsolateral aspect of 1 corpus cavernosum across the midline to the other side such that, as the knots were tied, the corpora rotated toward the dorsal midline. The knots were buried by apposition of the rotated corporal bodies. RESULTS: Excellent straightening of the phallus was achieved intraoperatively in these patients. CONCLUSIONS: The split and roll technique for correcting severe chordee does not require incisions into the corporal substance, involve use of grafts or cause shortening of the phallus. The neurovascular bundles are preserved and are not compressed by the rotational sutures. The surgeon may perform 1-stage reconstruction while achieving maximal penile length. PMID- 10458454 TI - Distal hypospadias repair by the modified Thiersch-Duplay technique with or without hinging the urethral plate: a near ideal way to correct distal hypospadias. AB - PURPOSE: Various techniques have been described to correct distal hypospadias but many are best suited to patients with subtle specific nuances of meatal and/or glanular configuration. We describe a simplified approach to these defects using the modified Thiersch-Duplay technique with or without hinging the urethral plate (the Snodgrass maneuver). MATERIALS AND METHODS: Between 1990 and 1997 we performed the modified Thiersch-Duplay repair as the primary procedure for repairing hypospadias in 197 boys. We initially performed urethral plate hinging (the Snodgrass maneuver) in 1994. By 1997, 82% of our patients were undergoing repair using the hinge technique. Urethroplasty coverage was provided by a second layer of adjacent local tissue or by a pedicle of subcutaneous tissue. RESULTS: Reoperation was required in 5 boys (2.5%) with fistula and 1 (0.50%) with urinary extravasation. A fistula developed in 4 of the 63 cases (6.4%) in which adjacent local tissue was used to cover the urethroplasty and in 1 of 130 (0.80%) in which a pedicle of subcutaneous tissue was used. Overall 97% of the boys had an excellent result requiring no secondary operative procedure. CONCLUSIONS: The modified Thiersch-Duplay technique with or without urethral plate hinging is reliable for correcting distal hypospadias. The risk of fistula is almost eliminated when a vascularized pedicle of subcutaneous tissue is used to cover the repair. This technique has virtually supplanted all other methods that we used in the past to correct distal hypospadias. PMID- 10458455 TI - Does tubularized incised plate hypospadias repair create neourethral strictures? AB - PURPOSE: Boys in whom hypospadias was repaired by tubularized, incised plate urethroplasty were evaluated to determine whether dorsal incision of the urethral plate results in strictures of the neourethra. MATERIALS AND METHODS: Postoperative followup of the initial 72 patients was reviewed and then attempts were made to contact these boys for delayed assessment more than 1 year after surgery. Periodic neourethral calibration was routinely done during year 1 postoperatively and at delayed followup. Urethroscopy was performed for a decreased urinary stream or during anesthesia for other procedures. Uroflowometry was done at the delayed assessment of toilet trained boys. RESULTS: The urethra was calibrated in 85% of the patients with no strictures noted. Urethroscopy in 10% of the boys demonstrated a healthy appearing neourethra. Uroflowometry was possible in 34% of the toilet trained patients with all results within the 95% confidence levels of standard curves for age. CONCLUSIONS: The dorsal relaxing incision of tubularized, incised plate hypospadias repair did not result in neourethral strictures. PMID- 10458456 TI - Medical management of phimosis in children: our experience with topical steroids. AB - PURPOSE: Circumcision has traditionally been regarded as primary therapy for persistent phimosis in boys. Recently groups in Europe and Australia have advocated the use of topical steroids as conservative treatment in children. We report our experience with this approach. MATERIALS AND METHODS: Between July 1997 and February 1998, 25 boys with a mean age of 8.3 years who presented to our clinic with phimosis were started on a topical steroid. After counseling the family regarding treatment options we prescribed a 1-month course of 0.05% betamethasone cream applied twice daily. RESULTS: Of the 25 patients 24 completed the treatment and were evaluated. A total of 16 boys (67%) had a normal appearing foreskin that was easily retracted, while in the remaining 8 the outcome was unsuccessful and circumcision was scheduled. CONCLUSIONS: Our study demonstrates that the application of topical steroids is a viable alternative for treating phimosis in children. Appropriate candidates for this therapy include boys older than 3 years who have persistent phimosis and no evidence of infection. PMID- 10458457 TI - Concealed penis in childhood: a spectrum of etiology and treatment. AB - PURPOSE: Concealed penis is an uncommon condition due to poor skin fixation at the base of the penis, cicatricial scarring after penile surgery and excessive obesity. The condition varies in severity and several surgical options are available, such as excision of previous scarring, degloving the penile shaft, reconstructing the penile shaft skin with flaps, fixing the penile skin at the penopubic and penoscrotal angles, and removing excess suprapubic fat. MATERIALS AND METHODS: We reviewed the records of 43 patients treated for concealed penis from 1993 to 1998. We categorized the cases as type 1-congenital concealed penis, type 2-concealed penis due to scarring from previous surgery and type 3-complex cases involving excessive obesity. Cases were reviewed in regard to surgical techniques and outcomes. We identified 18 type 1, 18 type 2 and 7 type 3 cases. Mean age of type 1 patients at surgery was 12.4 months with 1 patient presented at age 7 years. None had previously undergone penile surgery. All patients underwent complete penile degloving. To reconstruct the penile shaft flaps or Z plasties with penile skin were used in 12 patients and scrotal skin flaps were used in 2. In 12 patients the penile skin was fixed at the penoscrotal and penopubic angles to maintain penile length and in 2 excess fat was excised. Mean age of type 2 patients at surgery was 19.8 months. All had previously undergone surgery, including hypospadias in 1 and circumcision in 17. All patients underwent complete penile degloving and the cicatricial scar that trapped the penis was excised. Penile skin flaps and Z-plasties were used in 12 cases, scrotal skin flaps were used for reconstruction in 2 and skin grafting was done in 1. In 10 patients the penile skin was fixed with sutures to maintain penile length. Mean age of type 3 patients at surgery was 15.8 years. Of the 7 boys 6 had previously undergone penile surgery. All required extensive scar excision and complex reconstruction involving penile skin flaps in 3, scrotal flaps in 5 and penile skin fixation in 6. Excessive suprapubic fat was removed in 5 patients, of whom 3 underwent liposuction. RESULTS: Surgical results were uniformly good in type 1 patients except in 1 who was believed to have excessive suprapubic fat. Results were good in 14 of the 18 type 2 patients, although 2 retained excessive suprapubic fat and 2 had some unsightly scarring. No type 1 or 2 patient required additional surgery. Of the 7 type 3 patients 6 had a good result and required no additional surgery. One patient has recurrent concealed penis after 2 procedures and awaits additional surgery. CONCLUSIONS: Concealed penis has a varied etiology and requires a flexible surgical approach. The common surgical options in all cases include complete penile degloving, excising the scarring due to previous surgery, removing excess suprapubic fat, reconstructing the penile skin with local flaps, and fixing the penile skin at the penopubic and penoscrotal angles. PMID- 10458458 TI - Risk of contralateral hydrocele or hernia after unilateral hydrocele repair in children. AB - PURPOSE: Recent laparoscopic studies indicate a high incidence of a contralateral open internal ring in children undergoing unilateral hydrocele or hernia repair, raising the question of whether routine contralateral exploration should be done. Data on the long-term risk of clinical contralateral hernia or hydrocele after unilateral hydrocele repair are limited. To address this question we performed long-term followup in patients who underwent unilateral hydrocele repair. MATERIALS AND METHODS: We followed patients who previously underwent unilateral hydrocele repair performed by one of us before 1997. Patients were interviewed by telephone and encouraged to return to one of us or their pediatrician for evaluation. RESULTS: Of the 101 patients who fulfilled study inclusion criteria 85 who were 5 to 107 months old (median age 37) at the original surgery were successfully contacted, including 45 examined by one of us or a pediatrician and 40 followed by telephone interview only. Contralateral hydrocele or hernia developed in 6 of the 89 patients (7%) 6 to 15 months (median 12) postoperatively. The remaining 79 patients have been recurrence-free for 6 to 153 months (mean 44, median 37). Of the patients 5 of 32 are (15%) and 1 of 53 (2%) who underwent left and right hydrocele repair, respectively, had contralateral recurrence. CONCLUSIONS: The risk of a clinically evident contralateral hydrocele or hernia after unilateral hydrocele repair is approximately 7%. We do not recommend routine contralateral exploration in children undergoing unilateral hydrocele repair. PMID- 10458459 TI - Urodynamic findings in children with isolated epispadias. AB - PURPOSE: The exstrophy-epispadias complex represents a spectrum of anomalies with variable implications for upper and lower urinary tract function. Successful treatment of incontinence in this population is challenging and often elusive. To date few studies have focused on urodynamic findings specific to primary epispadias. MATERIALS AND METHODS: We retrospectively reviewed the records of patients presenting with primary epispadias in the absence of bladder exstrophy. We identified 18 boys and 12 girls, of whom 16 underwent formal urodynamic evaluation before (5), after (6) or before and after (5) surgical narrowing of the bladder neck. Bladder capacity was recorded as percent of expected capacity for age. Bladders were considered hypertonic when end filling pressure was greater than 20 cm. water. End filling pressure was defined as the pressure at which urinary leakage was noted or the patient had discomfort. Uninhibited contractions were considered significant when the amplitude was greater than 15 cm water. The ability of the detrusor to generate a voiding contraction at the end of the filling phase was also recorded. Urodynamic studies were performed with a balloon catheter occluding the incompetent bladder neck or with a standard urodynamic catheter after formal bladder neck repair. RESULTS: Before bladder neck repair average bladder capacity was 157 ml (range 55 to 450), corresponding to 52% (range 22 to 100) of expected capacity. Significant uninhibited contractions were noted in 2 patients. In 7 of the 9 evaluable patients (78%) a voiding contraction was generated. In children in whom urodynamic studies were done after surgery mean bladder capacity was 260 ml (range 77 to 660), corresponding to 76% (range 36 to 147) of expected capacity. Hypertonicity and hyperreflexia were noted in 3 and 2 patients, respectively. In 5 of the 11 cases (46%) a voiding bladder contraction was generated. Of the 5 patients who underwent urodynamic evaluation before and after bladder neck repair the percent increase in bladder capacity was significantly greater in the 2 boys (67 and 110%, respectively) than in the 3 girls (-20, 10 and 20, respectively). CONCLUSIONS: The most frequently noted urodynamic pattern before surgical correction of the bladder neck in patients with isolated epispadias is a low capacity, highly compliant bladder with minimal detrusor dysfunction. Postoperatively capacity increases, albeit to a greater extent in boys than in girls, and the incidence of detrusor dysfunction increases as well. In some patients classic bladder neck repair may result in detrimental bladder dynamics due to as yet poorly elucidated mechanisms. PMID- 10458460 TI - Male epispadias repair: surgical and functional results with the Cantwell-Ransley procedure in 40 patients. AB - PURPOSE: We present our experience using the Cantwell-Ransley epispadias technique, particularly focusing on postoperative anatomical and functional complications. MATERIALS AND METHODS: Between 1989 and 1997, 40 patients 1 to 28 years old underwent the Cantwell-Ransley technique for epispadias at our institution. The condition was isolated in 17 cases and associated with exstrophy in 23. Surgery involved a primary and secondary procedure in 29 and 11 patients, respectively. Cavernocavernostomy was performed in only 16 cases. RESULTS: At a mean followup of 3 years 18 patients (45%) had complications and needed further procedures, and 3 (7.5%) had major wound dehiscence. The complication rate was higher in the exstrophy than in the isolated epispadias group (65 versus 28%) and urethral complications were consistently associated with previous urethral plate sectioning. In 1 patient major and persistent loss of continence was probably related to prolonged transurethral bladder drainage. All 17 patients in the postpubertal group report erections, although 1 who did not undergo cavernocavernostomy still complains of dorsal curvature. In 36 patients (90%) a fully satisfactory anatomical and functional result was achieved. CONCLUSIONS: The Cantwell-Ransley technique of epispadias repair allows successful reconstruction in most patients. However, postoperative complications, some of which are serious, may develop that are more associated with previous procedures that compromise the blood supply to the urethral plate than the Cantwell-Ransley technique. PMID- 10458461 TI - Penile disassembly technique for epispadias repair: variants of technique. AB - PURPOSE: We present 2 variants of the penile disassembly technique for epispadias repair that refine some details of the Mitchell technique. In some cases the urethral plate retracts and shortens, and there may be poor vascularization at the most distal portion. In addition, when the neurovascular bundles of the separated hemicorporeal glanular bodies are intact, it is difficult to achieve excellent correction of dorsal chordee. MATERIALS AND METHODS: Between 1995 and 1998 we performed the modified Mitchell technique in 11 boys 2 to 14 years old using 1 of 2 variants. For variant 1 the hemiglans and urethral plate remain connected by a small tissue bridge to avoid shortening the urethral plate and ensure a better blood supply. For variant 2 each corporeal body is dissected from the glans cap and neurovascular bundle to achieve complete mobility. This procedure enables ideal mobility of the corporeal bodies as well as curvature repair. When corporeal rotation was unsuccessful, we corrected persistent dorsal chordee using the Ransley corporotomy with corporostomy in 2 patients and with dermal grafting in 1. RESULTS: Mean followup was 17 months (range 6 to 30). Dorsal curvature was corrected in all cases. Cosmetic appearance was good. Complications included meatal stenosis and urethral fistula in 1 case each. CONCLUSIONS: Our variants of epispadias repair may be good alternatives to the Ransley and Mitchell complete penile disassembly techniques. PMID- 10458462 TI - Endoscopic correction of vesicoureteral reflux in children using autologous chondrocytes: preliminary results. AB - PURPOSE: Previous approaches to the endoscopic correction of vesicoureteral reflux have used foreign bulking substances, raising concern regarding safety and long-term efficacy. We describe the results of a clinical trial using transurethral injection of autologous chondrocytes to correct vesicoureteral reflux in children. MATERIALS AND METHODS: A total of 29 children (46 ureters) with grades II to IV vesicoureteral reflux were treated at 2 sites. Each child underwent cystoscopy and ear cartilage biopsy at the initial setting. Chondrocytes were grown in culture for 6 weeks. Patients then returned for transurethral injection of chondrocytes into the bladder trigone to correct reflux. Ultrasound was performed 1 month and radionuclide cystography was done 3 months postoperatively to confirm reflux resolution. When reflux persisted, repeat treatment with stored chondrocytes was offered. RESULTS: Initial chondrocyte injection corrected reflux in 26 of the 46 ureters (57%), while secondary injection was successful in 12 of 19 (63%). Overall reflux was corrected in 38 of the 46 ureters (83%) and in 24 of the 29 patients (83%). There were no significant complications. CONCLUSIONS: Transurethral injection of autologous chondrocytes to correct vesicoureteral reflux in children appears to be an effective and safe technique that holds promise for treating this congenital abnormality in a minimally invasive fashion. PMID- 10458463 TI - No further development of renal scarring after antireflux surgery in children with primary vesicoureteral reflux: review of the results of 99mtechnetium dimercapto-succinic acid renal scan. AB - PURPOSE: Reports of previous studies using excretory urography indicate that significant numbers of new scars developed in 5 to 31% of cases and scarring progressed in 11 to 16% after antireflux surgery. We evaluated renal scarring after surgery using a more accurate method, that is dimercapto-succinic acid renal scintigraphy. MATERIALS AND METHODS: Between 1985 and 1997 antireflux surgery was performed for primary vesicoureteral reflux in 223 children at our hospital. Of these patients 45 boys and 29 girls with a median age of 74 months at surgery in whom preoperative and postoperative renal scans were available form the basis of our retrospective study. Renal scan was performed 0.3 to 58 months (median 2) preoperatively and 5.3 to 44 months (median 18.7) postoperatively. A total of 24 children (39 renal units) were followed further by additional scanning for as long as 25 to 120 months (median 43.2) postoperatively. Each scan was blindly reviewed for the size, number and zone location of cortical defects based on morphology. Interval changes were grouped into categories as improved, no change, progressed and new scar formation. RESULTS: Postoperatively there was no ureteral obstruction. Pyelonephritis developed in 2 patients (2.7%) and asymptomatic bacteriuria was noted in 35 (47.3%) postoperatively. Of the renal units 110 (86.6%) showed no change, while 15 (11.8%) were improved and 2 (1.6%) had progression. In no case was new renal scar formation observed. CONCLUSIONS: Contrary to most previous reports involving excretory urography, our results indicate no significant progression of renal scarring after antireflux surgery. In most cases postoperative followup renal scans demonstrated no significant morphological change. When there was change, it mostly involved the disappearance or decrease of renal scars. PMID- 10458464 TI - Quantitative nuclear cystography does not predict outcome in patients with primary vesicoureteral reflux. AB - PURPOSE: Quantitative nuclear cystography has been advocated as a tool for determining the prognosis in children with primary vesicoureteral reflux. We reviewed our data on this technique to assess its usefulness for predicting the outcome in this population. MATERIALS AND METHODS: We retrospectively reviewed the records of all patients with primary reflux in whom findings were positive on at least 2 nuclear cystograms at our institution between 1992 and 1997. Patients followed at least 3 years were stratified according to outcome. Unfavorable prognostic criteria included bladder volume at reflux onset 60% or less of total bladder capacity and calculated volume of reflux 2% or greater of bladder capacity. RESULTS: Of the 107 patients in our study 63 were followed for 3 years or longer, and reflux resolved in 17, was repaired in 24 and persisted in 22. Mean patient age at latest followup, duration of followup and number of cystograms did not significantly differ among groups. Intermittent reflux in 33% of the patients followed 3 years or longer was not associated with outcome or detrusor instability. Bladder and reflux volume varied and was nonpredictive in individuals. CONCLUSIONS: Quantitative nuclear cystography did not predict the outcome in patients followed for primary vesicoureteral reflux at a single institution for 3 years or longer. Intermittent reflux was common. These data suggest that nuclear cystography cannot be used to assess reliably the prognosis in individuals. Strong consideration should be given to using negative findings on 2 cystograms to confirm reflux resolution in patients at high risk. PMID- 10458465 TI - Urinary retention after bilateral extravesical ureteral reimplantation: does dissection distal to the ureteral orifice have a role? AB - PURPOSE: We evaluated the outcome and incidence of urinary retention after bilateral detrusorrhaphy using 2 modifications of the original Lich-Gregoir procedure. MATERIALS AND METHODS: We evaluated the charts of 220 patients who underwent correction of bilateral vesicoureteral reflux using the extravesical approach from January 1991 to December 1997. Inverted Y detrusorrhaphy was performed in 154 patients and the advancing suture modification was done in 66. RESULTS: The success rate using the advancing suture technique was 92.4 and 95.4% at 3 and 12 months, respectively. The Y detrusorrhaphy technique was successful in 91.6 and 97.4% of cases at 3 and 12 months, respectively. The difference in the techniques was not statistically significant. Urinary retention developed in 8.4% of the patients who underwent Y detrusorrhaphy compared to 15.2% of those who underwent the advancing suture technique (not statistically significant). However, patients with grades IV and V reflux, children younger than 3 years and boys had significantly (p <0.05) higher postoperative retention rates of 24.6, 35.6 and 20.3%, respectively, when all 220 patients were considered. CONCLUSIONS: Each modification of the original Lich-Gregoir technique is highly effective for treating bilateral vesicoureteral reflux and each is associated with a low rate of temporary urinary retention. Although there seems to be a lower incidence of retention with the Y detrusorrhaphy technique, this was not statistically significant. Because of the higher incidence of urinary retention in young children, boys and/or patients with high grade reflux, we recommend that physicians consider a longer period of catheterization regardless of the technique chosen. PMID- 10458466 TI - Cystography after the Cohen ureterovesical reimplantation: is it necessary at a training center? AB - PURPOSE: Reimplantation by the Cohen procedure has a low rate of recurrent reflux, although postoperative cystography is done routinely at most centers. According to the French training program for pediatric surgery and urology residents, reimplantation is the main pediatric urology procedure performed during residency. We determine whether it is necessary to perform postoperative cystography routinely and whether the fact that the procedure is done by a junior surgeon modifies management. MATERIALS AND METHODS: A total of 268 children with primary vesicoureteral reflux underwent ureteral reimplantation by the Cohen transtrigonal technique. Bilateral reimplantation was done in 97% of the cases. Reimplantation was performed by a surgery resident assisted by a clinical fellow or senior consultant surgeon in 37% of the cases. Routine cystography and renal ultrasound were done in all patients postoperatively. Followup ranged from 6 months to 5 years (mean 10 months). RESULTS: In 2 children (0.7%) with recurrent reflux surgery was not performed by a resident. One of the 2 children had asymptomatic persistent reflux and no further surgery was done. In the other child postoperative cystography was normal at 6 months. One year later she had acute pyelonephritis with recurrent unilateral reflux and underwent repeat reimplantation. CONCLUSIONS: Routine cystography is not necessary after bilateral Cohen reimplantation. Reflux recurrence is low even at a training center where surgery may be performed by junior surgeons. PMID- 10458467 TI - A multicenter outcomes analysis of patients with neonatal reflux presenting with prenatal hydronephrosis. AB - PURPOSE: Approximately 10 to 30% of prenatal cases of hydronephrosis result in the postnatal diagnosis of vesicoureteral reflux. Using a new generic prenatal postnatal data sheet developed by the Society for Fetal Urology the characteristics, natural history and outcome of prenatal hydronephrosis confirmed postnatally to be vesicoureteral reflux were documented at 3 centers. MATERIALS AND METHODS: We performed a retrospective multicenter review of Society for Fetal Urology data sheets completed for each patient in whom prenatal hydronephrosis was proved to be postnatal vesicoureteral reflux from 1993 to 1998. RESULTS: In 56 male and 15 female patients with prenatal hydronephrosis a total of 116 refluxing renal units were confirmed postnatally. Of the 116 renal units 112 were hydronephrotic prenatally. During gestation increased hydronephrosis was noted with voiding in 4 cases. Of the 112 hydronephrotic renal units only 26 ureters in 15 patients were seen prenatally. The obstetrician considered the diagnosis of vesicoureteral reflux in only 24% of the cases. Postnatally 116 refluxing renal units were identified. Initial postnatal ultrasound was normal in 25% of the cases. Bilateral reflux was present in 36 male and 9 female patients. In 10 of the 19 uncircumcised patients (53%) urinary tract infection developed despite antibiotic prophylaxis. In 15 of the 74 renal units with grades III to V reflux the condition resolved at an average patient age of 0.9 and 2.1 years in boys and girls, respectively. A total of 27 refluxing renal units were reimplanted. CONCLUSIONS: The majority of prenatal reflux occurs in boys, and it is high grade and bilateral. The data sheets designed by the Society for Fetal Urology are useful data collection instruments. The presentation and natural history of vesicoureteral reflux are different in male and female individuals. In a significant number of renal units high grade reflux resolves spontaneously. Early circumcision may decrease the incidence of breakthrough urinary tract infection in this subpopulation. In addition, the effective management of prenatally detected reflux depends on multispecialty communication. PMID- 10458468 TI - Ureteral reimplantation in infancy: evaluation of long-term voiding function. AB - PURPOSE: The advisability of early ureteral reimplantation in neonates and infants is controversial and to our knowledge long-term results are not available. We evaluated long-term voiding function after ureteral reimplantation in infancy. MATERIALS AND METHODS: Between 1984 and 1993, 32 children underwent ureteral reimplantation in year 1 of life at our institution. Long-term results were evaluated in regard to surgical outcome, voiding function, urinary tract infection and the need for prophylaxis. Analysis of voiding function included family interviews, uroflowmetry and post-void residual urine measurement. RESULTS: Complete long-term data were available in 14 girls and 7 boys. The diagnosis was primary bilateral and unilateral vesicoureteral reflux, and ureterovesical junction obstruction in 11, 4 and 6 cases, respectively. Of the 26 refluxing renal units overall disease was grade II in 3, III in 6 and IV to V in 17. Patient age at surgery was 0.6 to 12 months (mean 5.4) and followup was 5 to 13 years (mean 9.5). Tapered reimplantation was performed in 8 renal units using excisional or infolding in 5 and 3, respectively. There were no complications in 19 patients (94%), while 2 had postoperative reflux for which 1 underwent reoperation. In 13 patients voiding habits were normal, while 8 reported infrequent voiding (3 or fewer voids daily). In 19 of the 20 patients tested voided volume was appropriate, and the flow rate of 10.8 to 52.7 cc per second (mean 20.9) and post-void residual urine volume of 0 to 40 cc (mean 11) were considered normal. One patient with normal uroflowmetry had incomplete vesical emptying. CONCLUSIONS: Early reimplantation may result in a high technical success rate and low postoperative morbidity in infants. After long-term followup infrequent voiding was common but noninvasive assessment of bladder function revealed no significant abnormality in the majority of patients. PMID- 10458469 TI - Techniques of tissue handling and transfer. PMID- 10458470 TI - Does every patient with prenatal hydronephrosis need voiding cystourethrography? AB - PURPOSE: Prenatal ultrasound has allowed early identification of urinary tract abnormalities that may require urological followup or early intervention. While all children with prenatal hydronephrosis should undergo ultrasound within the first few weeks of life, we believe that there is a subset of postnatal hydronephrosis for which voiding cystourethrography can be avoided if careful observation is continued. MATERIALS AND METHODS: For 5 years 175 infants with a history of prenatal hydronephrosis were evaluated by ultrasound. Of 60 infants with less than Society for Fetal Urology grade II unilateral or bilateral hydronephrosis 44 underwent voiding cystourethrography as part of the early evaluation and 16 were observed without voiding cystourethrography. Four infants for whom we would routinely obtain voiding cystourethrography were excluded from study due to severe prenatal hydronephrosis, renal duplication, hydroureter, ipsilateral small or echogenic kidney and grade II or higher hydronephrosis. RESULTS: Voiding cystourethrography was positive in 6 of the 40 infants (15%) with less than grade II hydronephrosis. Of these cases 3 had grade III or higher vesicoureteral reflux and 1 with high grade reflux required reimplantation. None of the 16 patients followed without voiding cystourethrography has required further evaluation or intervention. In all patients with negative or no voiding cystourethrography parenchyma was preserved and hydronephrosis stabilized or resolved. CONCLUSIONS: Prenatal and postnatal ultrasound in infants should be used to guide further urological evaluation. Among infants with less than grade II hydronephrosis postnatally 15% had reflux on voiding cystourethrography, which is significantly higher than that reported among normal children (approximately 1%). However, none of the 16 infants observed without voiding cystourethrography on short-term antibiotic prophylaxis had deleterious renal events with 6 months to 4.5 years of followup. Therefore, we question the actual significance of the reflux detected in the first cohort of infants. Voiding cystourethrography can provide a definitive answer. However, we also believe that it is not absolutely mandatory based on the outcome in the observed group. With careful counseling and followup most patients with less than grade II hydronephrosis can be observed without urological sequela. PMID- 10458471 TI - Complications of voiding cystourethrography in the evaluation of infants with prenatally detected hydronephrosis. AB - PURPOSE: We determined complications in infants undergoing voiding cystourethrography as part of the evaluation for prenatally detected hydronephrosis. MATERIALS AND METHODS: We retrospectively reviewed the records of infants referred to our institution for the evaluation of prenatal hydronephrosis from 1992 to 1997. Infants with a prenatal history of bilateral hydronephrosis, bladder distention and oligohydramnios, oligohydramnios only or a prenatal abnormality involving any other organ system were excluded from study. Of 206 patients 129 male and 49 female infants underwent postnatal voiding cystourethrography at our institution. Chart review and a telephone interview with the parents were done to assess lower urinary tract infection, pyelonephritis, hospital admission for urosepsis, gross hematuria, urinary retention or skin rash. RESULTS: Postnatal voiding cystourethrography was normal in 138 patients but it diagnosed bilateral vesicoureteral reflux in 15, unilateral vesicoureteral reflux in 20, ureterocele in 4 and refluxing megaureter in 1. Of the 129 male infants evaluated 101 had undergone circumcision as a newborn, 14 were uncircumcised and the circumcision status of 14 was unknown. At voiding cystourethrography suppressive antibiotics were administered to 166 infants, 7 were not on suppressive antibiotics and antibiotic status was unknown in 5. No patient had a lower urinary tract infection, pyelonephritis or urosepsis. In addition, there were no episodes of urinary retention, gross hematuria or skin rash. CONCLUSIONS: While the reported rate of new or recurrent infection associated with voiding cystourethrography is as high as 6%, we did not identify any infectious or other complications in infants undergoing voiding cystourethrography for prenatal hydronephrosis. When properly performed, we believe that voiding cystourethrography is safe and presents little risk in these patients. PMID- 10458472 TI - Treatment of nocturnal enuresis with an ultrasound bladder volume controlled alarm device. AB - PURPOSE: Current treatment regimens for nocturnal enuresis are suboptimal. Medications such as desmopressin are efficacious for preventing the enuretic event but they offer little potential for a permanent cure and have side effects. Although the moisture alarm has good potential for a permanent cure, the child is mostly wet during treatment. Furthermore, the moisture alarm requires that the child make the somewhat remote association between the alarm event and a full bladder after the bladder has emptied. In this exploratory study bladder volume alarming, a new approach to treating nocturnal enuresis, was investigated. MATERIALS AND METHODS: A total of 40 sequential children 6 to 16 years old with nonorganic nocturnal enuresis who had had at least 1 wetting episode weekly for greater than a year were divided into 2 groups. Each child wore a modified PCI 5000 miniature bladder volume measurement instrument during sleep. An alarm sounded when bladder volume reached 80% of the typical daytime voided volume in group 1 and at 80% of the typical enuretic volume in group 2. Group 2 patients also performed daytime bladder retention exercises in regard to instrument measured bladder volume versus a progressing target volume. RESULTS: In groups 1 and 2 the mean dryness rate before study initiation versus during the study was 32.9 and 9.3 versus 88.7 and 82.1%, respectively. Nighttime bladder capacity increased 69% in group 1 and 78% in group 2, while the cure rate was 55% (mean treatment period 10.5 months) and 60% (mean treatment period 7.2 months), respectively. CONCLUSIONS: Bladder volume tracking seems to approach the goals of ideal treatment for nocturnal enuresis in that it prevents the enuretic event, appears to facilitate a permanent cure and is noninvasive. PMID- 10458473 TI - Staghorn calculi in children: treatment with monotherapy extracorporeal shock wave lithotripsy. AB - PURPOSE: We evaluated the effectiveness of monotherapy extracorporeal shock wave lithotripsy (ESWLT) for treating children with staghorn calculi. MATERIALS AND METHODS: From February 1992 to December 1997, 11 boys and 4 girls 14 months to 13 years old (median age 4 years) presented to our institution with staghorn calculi. In these patients ESWL was performed using a Siemens Lithostar-ULTRA with ultrasound stone localization and with intravenous sedation or without anesthesia. Using the Puigvert method the frequency and energy of the shock waves delivered were increased progressively to desensitize cutaneous nerve receptors, making the procedure less painful and improving stone fragmentation. RESULTS: Overall stones resolved in 11 of the 15 patients (73.3%) after an average of 2 ESWL sessions. Of the 11 patients 7 were stone-free after only 1 session, 2 with fragments less than 5 mm. required no further intervention, and 2 required additional surgery, including percutaneous nephrolithotomy to remove large residual stone fragments in 1 and open renal surgery to remove a cystine staghorn calculus in 1. Ureteral stents were not required in any patients. One case of post-ESWL fever resolved promptly with antibiotics. CONCLUSIONS: ESWL using the Siemens Lithostar-ULTRA is simple, effective and safe primary treatment in children with staghorn calculi. PMID- 10458474 TI - Strategies for managing upper tract calculi in young children. AB - PURPOSE: Pediatric urolithiasis is relatively uncommon and there is little information on the application of modern surgical procedures in young children. We present a single center experience with the surgical management of upper tract calculi in this age group. MATERIALS AND METHODS: We reviewed presentation, co morbidity, treatment, outcome and complications in all prepubertal patients who required surgical treatment for ureteral or renal calculi during a 4-year period. The series consists of 24 girls and 17 boys 17 months to 14 years old (mean age 7.5 years). A total of 26 children were anatomically normal, and 4 had myelomeningocele, 4 had ureteropelvic junction obstruction (in a pelvic kidney in 1), 2 had cloacal anomalies, 2 had vesicoureteral reflux, and 1 each had nonrefluxing megaureter, orthotopic ureterocele and a functioning renal transplant. RESULTS: Extracorporeal shock wave lithotripsy was performed in 24 patients. Stents or nephrostomy tubes were only used in the 4 patients who presented with pyonephrosis. Of the 41 cases 17 were rendered stone-free, 3 had a decreased stone burden and 4 were failures. Ureteroscopic extraction of distal ureteral calculi was successful in 11 of 12 children, of whom the youngest was 2.5 years old. No child had postoperative infection or evidence of ureteral obstruction. Stent placement facilitated stone passage or dissolution in 2 patients, a renal calculus was percutaneously extracted in 2 and 7 required open surgery, mostly for correcting simultaneous anatomical abnormalities or after minimally invasive surgery failed. Some metabolic abnormality was detected in 80% of the children tested. CONCLUSIONS: The surgical management of upper urinary tract calculi in young children parallels that in adults. Minimally invasive surgical methods may be safely used even in young infants. Most children do not need elective stenting before lithotripsy. Open procedures are still required in 17% of cases. The majority of children have definable metabolic abnormalities. PMID- 10458475 TI - The incidence of nephrolithiasis in patients with spinal neural tube defects. AB - PURPOSE: Bladder stones are common in patients with spinal neural tube defects but there are little data on the incidence of renal calculi in this population. We examined the incidence, nature and risk factors of nephrolithiasis in our clinic population of patients with neural tube defects. MATERIALS AND METHODS: We retrospectively reviewed the charts and radiological studies of 327 patients followed at our neural tube defects clinic with routine radiological imaging of the urinary tract. Additional confirmatory studies were performed when stones were noted. RESULTS: Renal calculi were identified in 20 patients with neural tube defects (6.1%). The incidence of nephrolithiasis increased with age. Renal stones were noted in 19 patients (10.7%) 12 years old or older. Management of the stones in these patients resulted in overall 53% stone-free and 87% recurrence rates after intervention. Major risk factors for new and/or recurrent renal stone formation were bacteriuria in 95% of the cases, lower urinary tract reconstruction in 80%, pelvicalicectasis in 70%, vesicoureteral reflux in 65%, a thoracic level spinal defect in 60% and renal scarring in 55%. CONCLUSIONS: These data suggest that there is a higher incidence of nephrolithiasis in patients with neural tube defects than in the general population and the risk of stone recurrence is also elevated. Most patients with stones had undergone lower urinary tract reconstruction. Other risk factors were bacteriuria, pelvicalicectasis, vesicoureteral reflux and a thoracic level neural tube defect. PMID- 10458476 TI - Pathophysiologic significance of host reactions in human cancer tissue: desmoplasia and tumor immunity. AB - Invasive growth of malignant cells, particularly carcinoma cells, induces host reaction within and around tumor tissue. Representatives of them are desmoplasia, angiogenesis and immune reactions. Desmoplasia, a process of fibrosis, is induced by activation of fibroblasts with increased production of matrix proteins and matrix degrading enzymes. Angiogenesis is prerequisite for the growth of solid tumor. Inhibition of this is now a target of cancer therapy. The present author has proposed a concept that tumor vessels are composed of nutrient vessels and immune/inflammatory vessels. The latter is similar to venules in inflammatory lesions expressing the cell adhesion molecules to facilitate the transmigration of inflammatory cells to the tissue. In colon cancer, venules distributed along the invasive margin correspond to these vessels, which express E-, and P selectins, and ICAM-1. These venules are considered to be an entry site of immune/inflammatory cells to cancer tissue. To further analyze immune mechanism, the present authors have confirmed that macrophages distributed along the invasive margin of colon cancer express costimulatory molecules B7.1/B7.2, which are required for the proliferation of T-cells. T-cells were co-localized with these cells. Clinicopathologic analysis confirmed that CD8+ T-cells distributed within cancer cell nest (intraepithelial) have the most significant impact on the patients' survival in colorectal cancer. These data suggest that various host reactions take place in the stroma of cancer tissue, which modulate the biologic behavior of cancer. PMID- 10458477 TI - Inhibitory effects of transforming growth factor-beta1 pretreatment on experimental pulmonary metastasis of MCS-1 Chinese hamster mesenchymal chondrosarcoma cells. AB - Recent studies have suggested that transforming growth factor(TGF)-beta1 acts as a multifunctional regulator of cell growth, and also modifies tumor progression and metastasis. In the present study, we investigated the effects of TGF-beta1 on the proliferation and experimental pulmonary metastasis of MCS-1. MCS-1 are undifferentiated type cloned tumor cells established from a mesenchymal chondrosarcoma which spontaneously occurred in the soft tissue of a female Chinese hamster. MCS-1 cells were pretreated with TGF-beta1 (0, 0.05, 0.5, 2, 10 ng/ml) for 72 hours in a medium containing 1% fetal bovine serum, then tested for in vitro growth by the MTT method, in vivo growth by subcutaneous inoculation into athymic nude mice (1 x 10(6) cells/mouse) and experimental pulmonary metastasis by injection into the lateral tail vein of athymic nude mice (5 x 10(4) cells/mouse). TGF-beta1 significantly inhibited in vitro growth of MCS-1, depending on its concentrations, and also experimental metastasis with maximal inhibition at 0.5 or 2 ng/ml treatment compared to untreated controls. TGF-beta1, however, was ineffective for in vivo subcutaneous growth of MCS-1. These results indicated that TGF-beta1 might be an inhibitor of metastasis of mesenchymal chondrosarcomas including other types of non-epitherial cartilage or bone formation tumors. PMID- 10458478 TI - Better-surviving liver grafts by the injection of anti-CD2 antibody: the important roles of host CD8+ and CD2+CD28+ T cells in chronic graft rejection and beta type platelet-derived growth factor receptor (PDGFR-beta) expression on apoptotic liver grafts. AB - Syngeneic liver grafts were implanted in the livers of 22 LEW/Sea strain rats. To prolong the graft survival, anti-CD2 monoclonal antibody (MAb) or anti beta type platelet-derived growth factor receptor (PDGFR-beta) antibody (Ab) was injected, or splenectomy was performed in the rats which were then followed until 10 to 11 weeks posttransplantation. The 22 rats with chronic graft rejection showed increased CD8a-like antigen (probably Fas ligand) on the peripheral blood T cells. All the liver grafts had both necrosis and apoptosis. The liver graft apoptosis was indicated by histopathological abnormalities, and by DNA strand breaks and hemosiderin depositions in the cytoplasm. PDGFR-beta expression in the apoptotic liver graft was demonstrated immunohistochemically. Among the 17 rats injected with anti-CD2 MAb, CD2 signaling on host T cells was effectively suppressed by the injection of anti-CD2 MAb in 4 rats with better-surviving liver grafts. In these 4 rats, CD28 antigen on thymic lymphocytes was down-modulated and high numbers (136-233-positive cells per lobe) of the epithelial reticular cells with apoptotic lymphocytes were counted. Anti-PDGFR-beta Ab caused high pulmonary secretions of growth factors and reticular fibrosis in the lungs of 5 rats injected with the Ab. Anti-PDGFR-beta Ab injection reduced the host cell apoptosis in the lung and thymus, but did not prolong the survival of liver grafts. In the 9 rats with both splenectomy and anti-CD2 MAb injection, pulmonary apoptosis was induced with the 6-16% reductions of CD4+ lymphocytes. Prolonged graft survival was observed in only one of the 9 rats. Anti-CD2 MAb was effective for prolonging the liver graft survival with suppressed CD28 antigen, but anti PDGFR-beta Ab and splenectomy were not. PMID- 10458479 TI - Administration of nerve growth factor, brain-derived neurotrophic factor and insulin-like growth factor-II protects phosphate-activated glutaminase in the ischemic and reperfused rat retinas. AB - Phosphate-activated glutaminase (PAG) activity decreases markedly in the early period of ischemia. The decrease of the enzyme activity is reversible if the ischemic period is relatively short, but it becomes irreversible after 90 minutes of ischemia. The deterioration is a functional damage of the retinas caused by ischemia. We studied effects of growth factors and neurotrophic factors on protection of PAG in the ischemic and reperfused rat retinas. Before ischemia, 1 microl of growth factors or neurotrophic factors (0.1 microg/microl for insulin like growth factor-I [IGF-I], insulin-like growth factor-II [IGF-II], brain derived neurotrophic factor [BDNF], nerve growth factor [NGF]; 1 microg/microl for basic fibroblast growth factor [bFGF]) were injected into the vitreous cavity of the left eyes of anesthetized Sprague Dawley rats. As a control, phosphate buffered saline was injected to the right eyes. To induce ischemia, we clamped left eyes for 90 minutes after bulbar conjunctival incision all around limbus. The rat retinas were homogenized with distilled water 1 day after reperfusion and used for PAG assay. Retinal ammonia concentration was also determined as a ischemic marker. About 80% decrease of retinal PAG activity and 50% increase of retinal ammonia concentration were observed after 90 minutes of ischemia and 1 day of reperfusion as compared with unoperated normal eyes. IGF-II, BDNF and NGF had protective effects on the retinal PAG activity, whereas IGF-I, bFGF, stable bFGF were less effective. In addition, IGF-II and BDNF suppressed elevation of retinal ammonia concentration. BDNF, NGF and IGF-II have marked effect on the protection of PAG activity in the ischemic and reperfused rat retinas, whereas bFGF, which is very effective for the protection of ischemic cell death, shows moderate effect. PMID- 10458480 TI - The effect of spinal instrumentation on lumbar intradiscal pressure. AB - The purpose of this study was to investigate the effect of spinal instrumentation on the intradiscal pressure (IDP) within the fixed motion segment. In vitro biomechanical testing was performed in six single functional spinal units of fresh calf lumbar spines using a pressure needle transducer. Various loads were applied by a materials testing system device. In addition to intact spine (control), anterior spinal instrumentation (ASI) and pedicle screw fixation (PS) constructs, as well as destabilized spine were tested. Relative to the control, the destabilized spine tended to have an increased IDP; by 15% in axial compression and by 9-36% in flexion-extension. Compared to the control, PS decreased the IDP by 23% in axial loading and 51% in extension loading and increased it by 60% in flexion for each loading. ASI decreased the IDP by 32% in flexion and 1% in extension. Lateral bending produced symmetrical changes of IDP in the control and destabilized spine, but no change in the PS construct. The IDP of the ASI construct was decreased by 77% in ipsilateral bending and increased by 22% in contralateral bending. These results demonstrated that eccentric loading from the spinal instruments increased IDP and significant disc pressure may still exist despite an increase in motion segment stiffness after lumbar stabilization. PMID- 10458481 TI - Localization of PDK-1 mRNA in the brain of developing and adult rats. AB - Gene expression for 3-phosphoinositide-dependent protein kinase-1 (PDK-1) in developing and adult rat brains was examined by in situ hybridization histochemistry. In embryonic days, the mRNA was evident throughout the entire neuraxis. The expression remained evident throughout the entire gray matters until postnatal day 7, and thereafter it decreased overall in the mantle and ventricular zones except for the cerebellar Purkinje and granule cell layers, the olfactory and hippocampal neuronal layers. The pattern of this gene expression is similar to those of for protein kinase B and class I phosphoinositide 3-kinases. PMID- 10458482 TI - Evaluation of a new anti-HCV-assay kit for anti-HCV screening in early childhood. AB - Sera of 20 children falsely identified as positive for hepatitis C virus antibody (Anti-HCV) by a second generation anti-HCV-assay kit (Imucheck-HCV Ab "Kokusai") were re-tested using a new third generation anti-HCV-assay kit (Imucheck x F-HCV C50 Ab "Kokusai"). Seventeen of the samples were reclassified as negative and only three remained positive. Changing well solids in the anti-HCV-assay kit from casein to bovine serum albumin appears to have improved the false-positive rate, most likely as a result of decreased non-specific adsorption of casein antibodies. PMID- 10458484 TI - Long-term azathioprine therapy in two children with steroid-dependent minimal change nephrotic syndrome. AB - Long-term azathioprine therapy as an alternative treatment to cyclophosphamide was done in 2 children with steroid-dependent minimal-change nephrotic syndrome (MCNS). They had already been treated with prednisolone, intravenous methyl prednisolone pulse therapy, cyclophosphamide and mizoribine. Although cyclophosphamide had been proved to be effective in maintaining their remission, the cumulative dose of the agent limited another course of cyclophosphamide therapy. Since ciclosporine therapy is much expensive, a trial of azathioprine (2 mg/kg per day) was started, and the therapy resulted in inducing sustained remission and reducing prednisolone. The patients were well tolerated the long term azathioprine therapy over a year. Although the efficacy of azathioprine in the management of childhood MCNS might be restricted, we therefore suggest that this agent should be reconsidered as an alternative treatment to cyclophosphamide. PMID- 10458483 TI - Point mutations in the steroid-binding domain of the androgen receptor gene of five Japanese patients with androgen insensitivity syndrome. AB - We analyzed the androgen receptor (AR) gene in five Japanese patients diagnosed with androgen insensitivity syndrome (AIS). All AR genes from the five patients had single-nucleotide substitutions, which introduced a premature termination codon in three patients (Gln640, Arg752, and Gln640 and Trp751), and a single amino acid substitution in two patients (Arg831 to Gln, and Leu812 to Phe). All the mutations occurred in the steroid-binding domain, comprising exons D through G. The three patients with the premature termination codon(s) and the one patient with Arg831Gln were clinically diagnosed as having complete AIS, while the patient with Leu812Phe had a partial form of AIS. Pubic skin fibroblasts from four of the five patients did not show detectable androgen binding. These data on mutations that have not been reported previously, provide valuable information for the further characterization of structural and functional relationships in the steroid-binding domain of the AR protein. PMID- 10458485 TI - Infantile pulmonary alveolar proteinosis with interstitial pneumonia: bilateral simultaneous lung lavage utilizing extracorporeal membrane oxygenation and steroid therapy. AB - An infant with refractory pulmonary alveolar proteinosis (PAP) associated with severe interstitial pneumonia is described. Although she was treated by bilateral simultaneous lung lavage utilizing extracorporeal membrane oxygenation and steroid therapy, she died of progressive respiratory failure 28 days after admission. Histologic examination of lung autopsy specimen showed only partial alveolar spaces to be filled with a dense PAS positive granular eosinophilic material and showed severe interstitial pneumonia with marked fibrosis of alveolar walls and interstitium. The lung lavage seemed to be effective for PAP because the effluent fluid sufficiently became clear and the PAS positive material was detected only in partial alveoli. The full venoarterial cardiopulmonary bypass with extracorporeal membrane oxygenation seemed to be very useful to support bilateral lung lavage for small infants. The refractory symptoms and failure of treatment were resulted from the association of severe interstitial pneumonia. In neonates or infants with PAP and severe interstitial pneumonia with poor response for steroid therapy, the lung transplantation should be considered. PMID- 10458486 TI - Prenatal confirmation of the translocation between chromosome 15 and Y-chromosome by fluorescence in situ hybridization. AB - A 30-year-old woman and her husband visited our hospital with habitual abortion as the complaint. Chromosome examination revealed a normal 46, XX for her and 46, XY, 15, der (15) t (Y; 15) (q12; p12) for him. After her pregnancy amniocentesis was performed. The karyotype was 46, XX, 15, der (15) t (Y; 15) (q12; p12) pat. ish der (15) (DYZ1+). A female baby was delivered. The growth of the baby was normal at 12 months of age. PMID- 10458487 TI - Hyaluronic acid suppresses the reduction of alpha2(VI) collagen gene expression caused by interleukin-1beta in cultured rabbit articular chondrocytes. AB - In order to investigate how alpha2(VI) collagen gene is regulated by inflammatory cytokines in cultured rabbit articular chondrocytes, we examined the effect of interleukin-1beta (IL-1beta) on this collagen mRNA expression. Polylayer cultures of chondrocytes were exposed to IL-1beta (0.1, 1, 10 ng/ml). Quantitative detection of specific mRNA for this collagen was carried out by reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, to investigate the effect of hyaluronic acid (HA) on alpha2(VI) collagen mRNA expression by IL-1beta chondrocytes were exposed to IL-1beta (10 ng/ml) in the presence of HA (0.01, 0.1, 1 mg/ml) with molecular weight of 900 kDa. Chondrocytes were also exposed to IL-1beta (10 ng/ml) in the presence of HA (1 mg/ml) with molecular weights of 200, 900 and 2000 kDa. Alpha2(VI) collagen mRNA expression was decreased significantly in chondrocytes cultured with 1 and 10 ng/ml of IL-1beta. However, the addition of both IL-1beta and HA (0.1, 1 mg/ml) or both IL-1beta and HA (1 mg/ml) with all the molecular weight significantly suppressed these reduced mRNA levels. No tendency for this suppression to depend on the molecular weight was observed. These results suggest that suppression of transcriptional activity for type VI collagen will be associated with the reduction of cartilage matrix tissue and that HA will be associated with the suppression of the effect of IL-1beta. PMID- 10458488 TI - Expression of MEF2 genes during human cardiac development. AB - To better understand the regulatory mechanisms in gene expression of human cardiomyocytes, we studied the expression of MEF2 genes encoding transcription factors during the course of cardiac development. Expression of all four MEF2 transcripts (MEF2A, MEF2B, MEF2C, and MEF2D) were detected in all developmental stage of the human heart, while Mef2b transcripts were down-regulated in mouse heart development. Although none of the MEF2 genes, besides mouse Mef2b, exhibited any remarkable quantitative change in their transcripts, qualitative changes in MEF2 transcripts were found during the course of cardiac development. In particular, MEF2D transcripts showed prominent changes by alternative splicing in the perinatal period. MEF2D transcripts containing the 21-base exon (exon b) were predominantly expressed after birth. At the same time, transcripts of the alpha myosin heavy chain (alphaMHC) gene increased after birth, as the splicing pattern in transcripts of the cardiac troponin T (cTnT) gene changed to decrease the transcripts of cTnT1 after birth. These changes seemed to be correlated with the alternative splicing changes of MEF2 genes, especially MEF2D. The alternative splicing as well as transcriptional regulation in MEF2 genes might be important for regulating the alphaMHC gene and the maturation of cardiomyocytes. PMID- 10458489 TI - Immunohistochemical characteristics of estrogen receptor alpha positive cells in glandular epithelium of the rat seminal vesicle. AB - Epithelial cells of the rat seminal vesicle stained positively for nuclear estrogen receptor alpha (ER alpha). We studied these cells using immunohistochemical means. We demonstrated in a previous study that some glandular epithelial cells of the seminal vesicles of immature castrated rats treated with estrogen for 1-2 weeks had multilayer features. The present study shows that these glandular epithelial cells are nuclear ER and basal cell specific cytokeratin (34betaE12) positive. These findings suggested characteristics of basal cells. Moreover, we demonstrated that these cells express transforming growth factor beta1 (TGFbeta1) as a result of castration and estrogen treatment. Our findings indicate that glandular epithelial cells with multilayer features, which stained positively for nuclear ER alpha have basal cell features and may play an important role in the expression of TGFbeta1 through an epithelial-stromal interaction. PMID- 10458490 TI - Endotoxin contamination in isolation of lamina propria mononuclear cells. AB - Because the beginning of extraction of lamina propria mononuclear cells is to obtain mucosal tissues that are exposed to luminal bacteria, the contaminated endotoxin in this step and/or the enzymes for mucosal digestion may activate mucosal macrophages and other cells. To address this issue, endotoxin levels in isolation solutions were evaluated during the extraction of lamina propria mononuclear cells from 8 control, 7 Crohn's disease and 8 ulcerative colitis specimens. Endotoxin levels were measured using Toxicolor system based on the limulus tests. Endotoxin levels were consistently below 500 pg/ml, and more importantly, these in enzyme digestion solutions were comparable among control, Crohn's disease, and ulcerative colitis. Therefore, comparative experiments using lamina propria mononuclear cells from these mucosae can be appropriately carried out, at least as far as in a comparable amount of contaminated endotoxin. However, careful consideration is required for the comparative and functional study using peripheral blood and lamina propria mononuclear cells. PMID- 10458491 TI - Point nucleotidic changes in both the RET proto-oncogene and the endothelin-B receptor gene in a Hirschsprung disease patient associated with Down syndrome. AB - A short-segment Hirschsprung disease (HSCR) patient associated with 21 trisomy showing point nucleotidic changes in both the receptor tyrosine kinase (RET) proto-oncogene and the endothelin-B receptor (EDNRB) gene is reported. A T to A heterozygous transition at the splicing donor site of the intron 10 in the RET proto-oncogene, and a G to A heterozygous substitution in non-coding region in the exon 1 of the EDNRB gene were observed. The familial analysis with these genes revealed that the origin of the former mutation was de novo and the latter one was maternal. No patient has been reported with two points mutations in different pathogenetically susceptible loci for HSCR. There is genetic evidence that the RET and EDNRB genes may interact in their susceptibility leading to HSCR. PMID- 10458492 TI - Relation with preoperative fructosamine and autonomic nerve function and blood pressure during anesthesia in diabetics: a retrospective study. AB - Many diabetics may have a high risk involving the cardiovascular system. In an attempt to predict the intraoperative risks of diabetics during anesthesia, we evaluated retrospectively the relationship among the biochemical assay or autonomic nerve function obtained as parts of the preoperative examination, and the blood pressure changes relating to the stimulation of intubation and extubation for anesthesia. In 40 diabetic surgical patients examined the biochemical assay (HbA1c, fructosamine level and blood glucose level) beforehand, the autonomic nerve function was quantified preoperatively by analysis of ECG R-R variability recorded in supine and subsequent standing position using an HRV analyzer, and some parameters of autonomic nerve function especially responsive sympathetic nerve activities were obtained. We assessed the correlation with systolic blood pressure changes in these cases at intubation for general anesthesia comparing to similar conditioned 40 non-diabetics. A diabetics with low vagal activity became larger systolic blood pressure afterdrop at tracheal intubation for anesthesia (r=0.513, p<0.001). Otherwise the blood pressure afterdrop at extubation became larger in a non-diabetics with high sympathetic activity (r=0.502, p<0.001). The preoperative fructosamine concentration in diabetics correlated positively with the responsive sympathetic nerve irritability index; "mRR(sup)-RRmin(std)" (r=0.432, p<0.05) and the responsive sympathetic nerve excitability index; "mRR(sup-std)" (r=0.448, p<0.05). However HbA1c had no correlation with these parameters of autonomic nerve function and blood pressure rise at tracheal intubation. Because of above correlation with blood pressure rise at intubation for anesthesia induction, the preoperative fructosamine examination and the responsive sympathetic nerve function test must be useful preoperative examination for detection of the unexpected heart events of diabetic patients during operation. PMID- 10458493 TI - Rise in plasma oxidized glutathione by experimental hypoglycemia. AB - Changes in plasma glutathione were investigated under hypoglycemic status. Twelve rabbits were randomly divided into hypoglycemic group (n=6) and saline-injected control group (n=6). Hypoglycemia was induced by intravenous injection of insulin as 10 U/kg and recovered by intravenous glucose injection after 60 minutes. In the control group, saline was intravenously injected in stead of insulin. Plasma levels of oxidized glutathione (GSSG) rose significantly (p<0.01) and remarkably decrease in plasma GSH/GSSG ratio (p<0.05) accompanying increase in serum enzymes in the hypoglycemic group. These results suggest that hypoglycemia might cause change in plasma GSSG which is related to increase of serum enzymes by hypoglycemia. PMID- 10458494 TI - Detection of serum antibody against arrestin from patients with acute disseminated encephalomyelitis. AB - In our previous study, we found the presence of serum autoantibody against arrestin in patients with multiple sclerosis (MS), while such serum autoantibody was not detected from patients with other neurological diseases and control subjects. We suggested that serum arrestin antibody titers may be useful for the diagnosis and evaluation of the disease's course. In the present study we examined sera from 7 patients, who were initially diagnosed as having acute disseminated encephalomyelitis (ADEM), for the presence of serum antibody against arrestin, in order to study the specificity of the serum antibody among demyelinated diseases. High titers were detected from 2 patients out of 7. One of the patients, a 4 year-old girl, presented with an additional neurological attack during the 6 months after the initial attack, resulting in change of diagnosis to MS. During her disease course the serum titers against arrestin fluctuated in correspondence with the disease's activity. These observations suggest that the presence of serum autoantibody against arrestin may be specific to MS and be helpful for differential diagnosis of ADEM and MS. PMID- 10458495 TI - Disseminated Fusarium infection identified by the immunohistochemical staining in a patient with a refractory leukemia. AB - The difficulty and uncertainty encountered in diagnosing a systemic mycosis often lead to a delay in starting antifungal therapy. We reported a disseminated infection of multiple fungal isolates including Fusarium species during donor leukocyte transfusion (DLT) after allogeneic bone marrow transplantation in a 20 year-old woman with a refractory leukemia. Skin lesions are the feature of Fusarium and occur in the early period of the infection. In this case, during immunosuppression state after DLT, she presented with the whole body ache and erythematous lesions which appeared rapidly on her trunk and extremities. While administration of amphotericin B was started, her condition was further deteriorated and she died. Autopsy materials revealed that she had multiple fungal infection with different isolates, including Aspergillus and Candida in the brain, lung and liver, but not in the skin. With the immunohistochemical staining with specific antibody, Fusarium or Aspergillus infection was identified from the biopsy skin or autopsy brain, respectively. This rapid and specific immunohistochemical method may be useful for the diagnosis and treatment of invasive fungal infection without delay. PMID- 10458496 TI - A case of interferon alpha-induced manic psychosis in chronic hepatitis C. AB - It is well known that mood disorder such as depression occasionally develops during interferon (IFN) therapy for chronic viral hepatitis. So far, however, IFN induced manic disorder has been rarely reported. We present a case of manic psychosis which developed during IFN treatment for chronic hepatitis C. A 35-year old man with chronic hepatitis positive for hepatitis C virus RNA in serum was treated with natural IFN alpha with a daily dosage of 5 million units. Six weeks later he complained of insomnia, and then became exhilarated, talkative, restless and aggressive. Since the mental state was compatible with manic disorder, IFN therapy was immediately ceased. Simultaneously, psychotropic drugs were administered. One week later, the psychiatric disturbances disappeared. He has been keeping his usual social interactions without the psychotropic drugs after that. It is suggested that manic psychosis happened secondary to IFN alpha treatment. PMID- 10458497 TI - Sensorineural hearing loss associated with Byler disease. AB - Progressive familial intrahepatic cholestasis, sometimes described as Byler disease, is a lethal liver disease and its inheritance is autosomal recessive. There is a previous report on the occasional association between this disease and sensorineural hearing loss without any audiological findings. We report here two siblings, an 18-year-old female and a 16-year-old male, suffering from Byler disease and hearing loss. Pure tone, Bekesy and speech audiometries and auditory brain stem response examination were performed. Audiometric data showed hearing characteristics of cochlear origin, high-frequency loss and progressiveness. This sensorineural hearing loss possibly results from a genetic mutation. The mechanism of cochlear disorder in patients with Byler disease is unknown, however, a novel gene responsible for deafness might be found to be related to Byler disease. PMID- 10458498 TI - Bridging the knowledge gap and communicating uncertainties for informed consent in cervical cytology screening; we need unbiased information and a culture change. PMID- 10458499 TI - A survey of biomedical scientists and consultant pathologists involved in the cervical screening programme. AB - The past decade has seen a transformation in the National Health Service Cervical Screening Programme (NHSCSP) largely driven by responses to various screening scandals. Independent inquiries into these laboratory failures have identified a number of contributing factors. The aim of this postal survey was to determine how widespread these factors are throughout screening laboratories in England. The results indicate that issues around recruitment and retention of staff, the training of pathologists and low morale appear to be widespread. However, the results do not indicate a widespread breakdown in working relationships between consultants and biomedical scientists, although there is room for improvement in their relationships with hospital management. Finally, there seem to be mixed messages about the commitment of consultants to the screening programme. PMID- 10458500 TI - Quality procedures in non-gynaecological cytology laboratories in England and Wales. AB - All non-gynaecological cytology laboratories in England and Wales (n = 212) were surveyed by telephone. The aim was to investigate what concepts of quality applied in this context and to establish what tools and techniques of quality improvement were used. The overall response was 146 (69%). The respondents mainly comprised NHS Trusts and University Departments. The study showed that there was a diverse approach to quality. All types of quality assurance and customer focus procedure questioned were undertaken but to a varied extent; three laboratories (2%) used a complete range and three (2%) used no procedure at all. Accreditation was associated with staffing adequacy and use of surveys, but not quality assurance (QA) or user focus. Laboratories with a high priority for quality performed more QA and reported a higher staffing adequacy. Critical incident analysis was dependent on workload. Computerization did not affect quality procedures and involvement in the Breast Screening Programme did not result in different quality measures. The time since last update was independent of all factors and external quality assurance (EQA) was not widely available. The study suggested that an integrated approach to quality had not been adopted in English and Welsh cytology laboratories and that there may be a need for a more strategic approach with greater availability of EQA, guidelines on quality tools, closer linkage of accreditation and quality procedures and the production of minimum and ideal standards. The ideal standard could be the complete range of procedures, and the minimum standard could comprise those processes in most frequent use, i.e. critical incident analysis, correlation methods, action on information, analysis of what is done with diagnostic information, a complaints procedure and customer surveys. PMID- 10458501 TI - Radial scar/complex sclerosing lesion--a problem in the diagnostic work-up of screen-detected breast lesions. AB - During the first 7.5 years of breast cancer screening in South Australia, 88 radial scar/complex sclerosing lesions were among the mammographically detected abnormalities. A false-positive cytological diagnosis by fine needle biopsy was given in three of 69 such cases with satisfactory smears, a false-positive rate of 4.3% in this particular group. A review of the smears suggested that the false positive diagnoses could have been avoided if a total or near total absence of a benign epithelial component had been included among the criteria for a malignant diagnosis. However, even after review, atypia was considered sufficiently worrying to be reported as a suspicion of malignancy in 7% of cases. PMID- 10458502 TI - Single-strand conformational polymorphism (SSCP)-detected p53 gene mutations are a less sensitive marker of malignancy in pleural fluids than p53 immunostaining. AB - p53 immunostaining has been advocated as a marker of malignancy in pleural biopsies and serous fluids. The object of this study was to compare the sensitivity and specificity of p53 immunostaining for the detection of malignant cells in pleural fluids with a technique designed to detect p53 gene mutations in exons 5, 6, 7 and 8 by SSCP and nucleotide sequencing. Five out of eight pleural fluids containing adenocarcinoma showed p53 immunostaining and two of these also showed polymorphisms on SSCP and a mutation on sequencing. None of the 10 benign pleural fluids showed immunostaining for p53 or polymorphisms on SSCP. We believe that the poor sensitivity of p53 gene mutation by SSCP is mainly due to DNA from the background reactive cells 'swamping' the mutant DNA. We do not advocate its use as a diagnostic aid. PMID- 10458503 TI - Fine needle aspiration (FNA) cytology of pilomatrixoma: report of 14 cases and review of the literature. AB - We describe 14 cases of pilomatrixoma which were diagnosed preoperatively on FNA cytology. In contrast to the literature, our series showed male preponderance and some of the tumours in unusual locations such as thigh and breast. In three patients the tumours were larger than 5 cm. The clinical diagnosis varied from tumour not otherwise defined to sarcoma. The cytologic presentation had features which allowed a correct diagnosis in all cases and included basaloid cells surrounded by delicate pink fibres, shadow cells, giant cells, naked nuclei and calcium deposits. It is concluded that the FNA cytology of pilomatrixoma is characteristic and will allow a conclusive diagnosis even in cases with an aberrant clinical presentation. PMID- 10458504 TI - Malignant pilomatrixoma: a case report with fine needle aspiration (FNA) cytology. PMID- 10458505 TI - Xanthogranulomatous sialadenitis following diagnosis of Warthin's tumour: a possible complication of fine needle aspiration (FNA). PMID- 10458506 TI - Aspiration cytology of a pleomorphic lipoma--a cautionary note. PMID- 10458507 TI - Pseudomalignancy in a hydrocele fluid. PMID- 10458508 TI - The audit of smear takers by personal performance profiles? PMID- 10458509 TI - Training in cervical cytology. PMID- 10458510 TI - Des-Arg9-bradykinin metabolism in patients who presented hypersensitivity reactions during hemodialysis: role of serum ACE and aminopeptidase P. AB - Bradykinin (BK) has been proposed as the principal mediator of hypersensitivity reactions (HSR) in patients dialyzed using negatively charged membranes and concomitantly treated with angiotensin-converting enzyme (ACE) inhibitors. We investigated the metabolism of exogenous BK added to the sera of 13 patients dialyzed on an AN69 membrane with a history of HSR (HSR+ patients) and 10 others who did not present such a reaction (HSR- patients) while dialyzed under the same conditions. No significant difference in the t1/2 of BK was found between the patient groups. However, the t1/2 of generated des-Arg9-BK was significantly increased (2.2-fold) in HSR+ patients compared to HSR-subjects. Preincubation of the sera with an ACE inhibitor (enalaprilat) significantly increased the t1/2 of both BK and des-Arg9-BK in both groups. There was no significant difference between the groups with respect to the t1/2 of BK, but there was a significantly greater increase (3.8-fold) in the t1/2 of des-Arg9-BK in HSR+ patients compared to HSR-subjects. The level of serum aminopeptidase P (APP) activity showed a significant decrease in the HSR+ sera when compared to HSR-samples. In HSR- and HSR+ patients, a significant inverse relation (r2 = 0.6271; P < 0.00005) could be calculated between APP activity and des-Arg9-BK t1/2. In conclusion, HSR in hemodialyzed patients who are concomitantly treated with a negatively charged membrane and an ACE inhibitor can be considered as a multifactorial disease in that a decreased APP activity resulting in reduced degradation of des-Arg9-BK may lead to the accumulation of this B1 agonist that could be responsible, at least in part, for the signs and symptoms of HSR. PMID- 10458512 TI - Bombesin injection into the central amygdala influences feeding behavior in the rat. AB - The present study was performed to determine whether low doses (10 or 40 ng) of bombesin microinjected into the amygdala could modify solid food intake. Forty ng of bombesin in 24 h deprived rats caused transient inhibition of food intake. This inhibitory effect was eliminated by prior bombesin antagonist treatment. A series of quantitative behavioral tests indicated that low doses of bombesin application specifically reduced food intake without altering the behavioral pattern or influencing the body temperature. The present results suggest, that bombesin-like peptides may act as a satiety signal in the central part of the amygdala. PMID- 10458511 TI - Cloning of a second proopiomelanocortin cDNA from the pituitary of the sturgeon, Acipenser transmontanus. AB - A recent study on the pituitary of the sturgeon, Acipenser transmontanus, resulted in the cloning of a cDNA that codes for the prohormone, proopiomelanocortin (POMC). This cDNA is designated sturgeon POMC A. Subsequent analysis of the sturgeon pituitary uncovered a second distinct POMC cDNA (sturgeon POMC B). In both sturgeon POMC cDNAs the open reading frame is 795 nucleotides in length. However, the two sturgeon POMC cDNAs differ at 26 amino acid positions in the opening frame. In addition, the 2 forms of POMC differ at 45 nucleotide positions within the open reading frame. The number and types of point mutations are compared in the 2 sturgeons POMC cDNAs, and the origin of the two POMC genes is discussed. PMID- 10458513 TI - Cyclic estradiol treatment phasically potentiates endogenous cholecystokinin's satiating action in ovariectomized rats. AB - The influence of ovarian cycling and of exogenous estradiol on the cholecystokinin (CCK) satiety-signalling system was investigated in intact and ovariectomized Long-Evans rats, respectively. Intraperitoneal injection of 1 mg/kg devazepide, the most potent and selective CCK(A) receptor antagonist, increased test meal size during estrus, but not during diestrus, confirming the influence of hypothalamic-pituitary-gonadal function on CCK satiety in intact rats. Devazepide was then tested in ovariectomized rats that received chronic cyclic estradiol (2 microg estradiol benzoate on Tuesday and Wednesday each week) or oil treatment. Devazepide did not increase meal size in estradiol-treated rats on Tuesday, prior to estradiol treatment, compared to oil-treated rats, but did selectively increase meal size on Friday, late in the estradiol replacement cycle, compared to Tuesday, early in the cycle. These results suggest that a phasic potentiation of the endogenous CCK satiety-signalling system is part of the mechanism for the decrease in meal size in female rats during estrus. PMID- 10458514 TI - Endogenous cholecystokinin's satiating action increases during estrus in female rats. AB - Food intake and meal size are reduced in female Long-Evans rats during estrus. To investigate the contribution of the satiating action of endogenous cholecystokinin (CCK) to this, rats were injected with 1 mg/kg of the potent, selective CCK(A) receptor antagonist, devazepide, during diestrus, when meal size is maximal, and during estrus, when it is minimal. Devazepide increased spontaneous food intake and meal size during estrus, but not during diestrus. Meal frequency was not affected by devazepide treatment. These results indicate that the potency of the CCK satiety-signaling system increases during estrus. PMID- 10458516 TI - Nociceptin/orphanin FQ increases blood pressure and heart rate via sympathetic activation in sheep. AB - This study was undertaken to examine the cardiovascular effects of nociceptin/Orphanin FQ (OFQ). Nociceptin/OFQ (10-300 nmol/kg, IV) stimulates an increase in mean blood pressure (MBP) and heart rate (HR) in chronically catheterized sheep. Pretreatment with phenoxybenzamine (5 mg/kg) attenuated the pressor response, consistent with sympathetically mediated vasoconstriction. Furthermore, the lack of a reflex bradycardia suggests either blunting of the baroreflex by nociceptin/OFQ or direct beta-adrenergic activation. The bradycardic response to norepinephrine (0.6 microg/kg, IV) remained intact after nociceptin/OFQ administration, demonstrating that nociceptin/OFQ does not blunt the baroreflex. Additionally, the increase in HR was completely reversed by pretreatment with propranolol. These data suggest that nociceptin/OFQ plays a role in cardiovascular regulation via sympathetic activation. PMID- 10458515 TI - Monitor peptide binding sites are expressed in the rat liver and small intestine. AB - 125I-monitor peptide binding was performed using frozen sections of the rat liver and gut and visualized using autoradiography. Saturable binding was observed in unidentified single cells in the liver and in the mucosa of the small intestine. Epidermal growth factor (EGF) and GTPgammaS did not inhibit 125I-monitor peptide binding indicating that the binding sites are not EGF receptors or G protein coupled receptors. The liver binding site exhibited an affinity 3.7-4.4-fold higher than those in the small intestine. It has been established that intraluminal monitor peptide releases cholecystokinin from the small intestine. The present results indicate that monitor peptide may also have liver associated functions. PMID- 10458517 TI - Chimeric peptide of Met-enkephalin and FMRFa induces antinociception and attenuates development of tolerance to morphine antinociception. AB - A synthetic chimeric peptide of Met-enkephalin and FMRFamide (YGGFMKKKFMRFa), based on MERF was synthesized. This peptide was tested for possible antinociceptive effects using the tail flick test in mice. The effect of the chimeric peptide on morphine antinociception and development of tolerance to the antinociceptive action of morphine was also investigated. The chimeric peptide produced significant, dose-dependent antinociception (40, 60 and 90 mg/kg) in the tail flick test. Pretreatment with naloxone (5 mg/kg, IP) significantly attenuated the antinociceptive effect induced by the chimeric peptide (90 mg/kg, IP), indicating involvement of an opioidergic mechanism. In combination experiments with morphine, the antinociceptive dose of the chimeric peptide (60 mg/kg, IP) potentiated morphine (7 mg/kg, IP) antinociception. A low dose of the chimeric peptide (10 mg/kg, IP), that did not produce significant antinociception on its own, also potentiated morphine antinociception. In the tolerance studies, male albino mice received twice daily injections of morphine (20 mg/kg, IP) followed by either saline (0.1 ml) or chimeric peptide (80 mg/kg, IP) for a period of 4 days. A control group received twice daily injections of saline (0.1 ml) for the same period. When tested on Day 5, tolerance to antinociceptive action of morphine (15 mg/kg, IP) was evidenced by decreased response in chronic morphine plus saline treated mice compared to control group. Concurrent administration of chimeric peptide (80 mg/kg, IP) with morphine significantly attenuated the development of tolerance to the antinociceptive action of morphine. The preliminary results of this study demonstrate that peripherally administered chimeric peptide can produce dose dependent, naloxone reversible, antinociception; potentiate morphine antinociception and attenuate morphine tolerance, indicating a possible role of these type of amphiactive sequences in antinociception and its modulation. These chimeric peptides may also prove to be useful tools for further ascertaining the role of FMRFa family of peptides in mechanisms leading to opiate tolerance and dependence. PMID- 10458518 TI - Tyr-W-MIF-1-induced conditioned place preference. AB - Based on the evidence that Tyr-Pro-Trp-Gly NH2 (Tyr-W-MIF-1) produced dose dependent, mu-opiate agonistic/antagonistic effects, we investigated whether Tyr W-MIF- exhibited similar properties in the conditioned place preference (CPP) test. To examine the opiate agonistic effects on CPP, rats were conditioned with alternating ICV injections of saline and Tyr-W-MIF-1 (0 or 200 microg). This procedure resulted in Tyr-W-MIF-1-induced CPP. To examine the opiate antagonistic properties of low doses of Tyr-W-MIF-1, morphine-induced CPP was challenged with Tyr-W-MIF-1 (0, 25, 50, or 100 microg). Morphine-induced CPP was not affected by Tyr-W-MIF-1 at these doses. These findings show that in the CPP test Tyr-W-MIF-1 produced opiate agonistic effects at the high dose and was without opiate antagonistic properties at lower doses. PMID- 10458519 TI - Atrial natriuretic peptide relaxes arterial basal tone induced by coarctation hypertension. AB - We have investigated the vasorelaxant effect of atrial natriuretic peptide (ANP) on isolated non-contracted aorta from coarctation hypertensive rats (HR) and the role of endothelium in this vasorelaxant action. After 7-14 days of surgery, mean blood pressure was higher (P < 0.01) in HR compared with sham operated rats (SR), used as the control. ANP (10(-6) mol/l) significantly lowered basal tone in previously unstimulated HR thoracic aortic rings; however, it had no effect in HR abdominal aorta or in SR abdominal and thoracic aorta. Endothelial destruction potentiated the vasorelaxant effect of ANP on basal tone in HR thoracic aorta. A similar potentiation of the ANP-response was observed by pre-treatment with N(G) nitro-L-arginine methyl ester (L-NAME, 3 x 10(-4) mol/l) or methylene blue (2 x 10(-5) mol/l) in unrubbed HR thoracic aorta. Treatment with calcium-free Krebs + EGTA (2 x 10(-3) mol/l) + sodium nitroprusside (10(-5) mol/l) or calcium-free Krebs significantly decreased basal tone and abolished ANP-response. These effects were observed only in HR thoracic aorta. Similarly, staurosporine (10(-7) mol/l) and calphostin C (10(-6) mol/l), inhibitors of protein kinase C (PKC), diminished basal tone and abolished the ANP-response in HR thoracic aorta. Acetylcholine (10(-6) mol/l) had a small but significant action on the basal tone of unrubbed HR thoracic aorta. These results demonstrate that ANP has a vasorelaxant effect on aortic basal tone when the vessel is exposed to high blood pressure. Inhibition of ANP effects on basal tone by calcium-free Krebs and PKC antagonists suggests that the HR aorta increases Ca2+-active tone, that modifies the response of vascular smooth muscle to the vasodilating hormone ANP. PMID- 10458520 TI - Potentiation of the hypotensive effect of bradykinin by angiotensin-(1-7)-related peptides. AB - In this study, we evaluated the bradykinin potentiating activity and ACE inhibitory activity of several Ang-(1-7)-related peptides: Ang-(2-7), Ang-(3-7), Ang-(4-7), Ang-(1-6), Ang-(1-5) and the selective antagonist of Ang-(1-7): D [Ala7]Ang-(1-7) (A-779). In vivo experiments were performed in freely moving Wistar rats. ACE activity was evaluated by a fluorometric assay in rat plasma using Hip-His-Leu as a substrate. Intravenous injections of Ang-(1-7) (2.2 nmol) transformed the effect of a single dose of bradykinin (1 nmol) into the effect produced by a double dose. A similar bradykinin potentiating activity was demonstrated for Ang-(2-7) and Ang-(3-7). On the other hand, Ang-(1-5), Ang-(1 6), Ang-(4-7) and A-779 did not change the hypotensive effect of bradykinin in doses ranging from 8 up to 25 nmols. The hypotensive effect of bradykinin was increased by intravenous infusion (0.3 ng/min) of Ang-(1-7) > Ang-(2-7) > Ang-(3 7). Conversely, Ang-(1-5), Ang-(1-6), Ang-(4-7) or A-779 did not change the hypotensive effect of bradykinin. ACE inhibition with Ang-(1-7) related peptides occurred in the order: Ang-(2-7) > or = Ang-(3-7) > Ang-(1-7) [>>] Ang-(1-5) > Ang-(4-7) > or = Ang-(1-6) > or = A-779. A-779 in concentrations up to 10(-5) M did not change the ACE inhibitory activity of Ang-(1-7). These results suggest that Ang-(1-7), Ang-(2-7) and Ang-(3-7) can modulate bradykinin actions in vivo. More important, our data pointed out that alternative mechanisms besides interaction with ACE are required to explain the bradykinin potentiating activity of Ang-(1-7). PMID- 10458521 TI - Developmental changes of hypothalamic, pituitary and striatal tachykinins in response to testosterone: influence of prenatal melatonin. AB - Substance P (SP) and neurokinin A (NKA), members of the family of mammalian tachykinins, are involved in the regulation of many physiological functions and are widely distributed in mammalian tissues. In this report, the effects of prenatal melatonin on the postnatal developmental pattern of NKA, and SP, and on testosterone secretion were investigated. Also, tachykinin response to the administration of testosterone propionate (TP) was studied. The brain areas studied were medio-basal-hypothalamus, pituitary gland and striatum. Male rat offspring of control or melatonin treated mother rats were studied at different ages of the sexual development: infantile, juvenile or prepubertal periods, and pubertal period. Both groups received exogenous TP (control-offspring+TP and MEL offspring+TP), or the vehicle (control-offspring+placebo and MEL offspring+placebo). Hypothalamic concentrations of all peptides studied in control-offspring+placebo remained at low levels until the juvenile period, days 30-31 of age. After this age, increasing concentrations of these peptides were found, with peak values at puberty, 40-41 days of age, then declining until adulthood. In the MEL-offspring+placebo a different pattern of development was observed; hypothalamic concentrations of NKA and SP from the infantile period until the end of juvenile period were significantly higher than in control offspring+placebo. TP administration exerted a more marked influence on MEL offspring than on control-offspring and prevented the elevation in tachykinin concentrations associated with prenatal melatonin treatment. TP administration to control-offspring resulted in significantly reduced (P < 0.05) tachykinin concentration only at 40-41 days of age, and increased (P < 0.01) during infantile period as compared to control-offspring+placebo. Pituitary NKA concentrations were lower than in the hypothalamus. In control-offspring+placebo pituitary NKA levels did not show significant changes throughout sexual development. A different developmental pattern was observed in MEL offspring+placebo, with significantly increased (P < 0.05) pituitary NKA concentrations at 35-36 days of age than in control-offspring+placebo. TP administration to control-offspring influenced pituitary NKA levels at the end of the infantile and pubertal periods, showing at both stages significantly higher (P < 0.05) NKA levels as compared to control-offspring+placebo. NKA levels in MEL offspring+TP were only affected at 21-22 days of age, showing significantly increased (P < 0.01) values as compared to MEL-offspring+placebo. Striatal tachykinin concentrations in control-offspring did not undergo important modifications throughout sexual development, but during the prepubertal period they started to increase. Maternal melatonin and TP injections produced short lived alterations during the infantile period. The results showed that prenatal melatonin delayed the postnatal testosterone secretion pattern until the end of the pubertal period and postnatal peptide secretion in brain structures. Consequently, all functions depending of the affected areas will in turn, be affected. PMID- 10458522 TI - Corticotropin-releasing factor (CRF) induced anorexia is not influenced by a melanocortin 4 receptor blockage. AB - CRF and melanocortin (MSH/ACTH) peptides share a number of central effects including anorexia and grooming. The effects of CRF may be secondary, due to CRF's effects on melanocortin peptide release. We investigated if the newly discovered selective melanocortin 4 receptor antagonist HS014 could influence CRF induced anorexia and grooming. The data show that ICV administration of CRF (3 mg/rat), significantly reduced food intake, feeding time and feeding episodes whereas it increased grooming time and grooming episodes. HS014 (5 mg/rat), that previously has been shown to antagonize the anorectic effect and the excessive grooming induced by alpha-MSH, did however not influence any of the behavioral effects induced by CRF when the peptides were administered together. The data indicate that the anorectic and grooming effects of CRF are independent of pathways involving the MC4 receptors. These data suggest that the anorectic and grooming effect of CRF are not due to a secondary effect caused by increase in release of melanocortins acting on the central MC receptors. PMID- 10458523 TI - Fluorescent and biotinylated probes for B2 bradykinin receptors: agonists and antagonists. AB - Peptide ligands carrying additional reporter groups are valuable research tools to facilitate biochemical and pharmacological studies of G protein-coupled receptors. B2 bradykinin receptors, widely distributed in mammalian tissues, regulate many physiological systems and are therapeutic targets. Acylation of the amino-terminus of bradykinin (BK) and a B2a-selective antagonist produced ligands derivatized with biotinamidocaproate or 7-Amino-4-methylcoumarin-3-acetate. These fluorescent and biotinylated peptides bound with high affinity to bovine and rodent B2 receptors. Analysis of second messenger production confirmed that fluorescent and biotinylated analogs of BK were B2 receptor agonists whereas derivatives of DArg0[Hyp3,DPhe7,Leu8]BK were BK receptor antagonists. The complimentary properties of these selective receptor probes will be useful in studying B2 receptor localization, expression and desensitization. PMID- 10458524 TI - cAMP and in vitro inotropic actions of secretin and VIP in rat papillary muscle. AB - Secretin and VIP stimulate cardiac adenylyl cyclase activity and exert a positive inotropic action in several mammalian species. This study examined positive inotropic activity and cAMP levels in rat papillary muscle. Isoproterenol and secretin increased contractions by 150+/-31% and 129+/-27%, respectively. VIP increased contraction by 30+/-21% only at 10 microM. Isoproterenol significantly increased cAMP levels by 82%, whereas increases by secretin (58%) and VIP (56%) were not significant. These results are consistent with reports that secretin and VIP stimulate cardiac adenylyl cyclase in the rat, but suggest that cAMP tissue levels cannot totally explain the positive inotropic responses to secretin and VIP. PMID- 10458525 TI - Role of polypeptides in the treatment and diagnosis of osteoporosis. AB - Osteoporosis is a common disorder characterized by reduced bone mineral density, deterioration of the microarchitecture of bone tissue and increased risk of fracture. The aim of treatment of osteoporosis is to maintain and, ideally, to restore bone strength safely. In recent years the role of polypeptide growth factors in bone metabolism has begun to appear. It has been proposed that alterations in the expression or production of growth factor can modulate the proliferation and activity of bone forming cells. In this direction, the role of structurally diverse peptides for the management and diagnosis of osteoporosis has attracted the attention of many investigators. This paper reviews numerous findings concerning the use of polypeptides, hormones, and growth factors, for the management of osteoporosis. Many of the compounds mentioned here are experimental prototypes of new therapeutic classes. Though it is unlikely that some of the compounds may ever be used clinically, development of safe and efficacious agents in each class will define the future course of therapy for osteoporosis. PMID- 10458526 TI - Pregnancy outcome following first-trimester exposure to zopiclone: a prospective controlled cohort study. AB - BACKGROUND & AIM: Zopiclone, a cyclopyrrolone derivative, is a short-acting hypnotic. To date, no published data exist regarding human pregnancy experience with zopiclone. The purpose of this study was to compare pregnancy outcome following first-trimester exposure to zopiclone with that of a matched control group of women, who were counseled for nonteratogenic exposure. METHODS: The Motherisk Program, the Toronto Teratogen Information Service, prospectively collected and followed up 40 women exposed to zopiclone during pregnancy. Pregnancy outcome was compared with that of a matched control group of women, who were counseled for nonteratogenic exposure. RESULTS: There was no increase in the rate of major malformations (0 of 31 [0%] for zopiclone vs. 1 of 37 [2.7%] for nonteratogenic controls; p = 1). CONCLUSIONS: Our study, which is the first cohort on zopiclone use during embryogenesis, albeit small, suggests that zopiclone does not appear to be a major human teratogen. Larger studies are needed to establish its safety during pregnancy. PMID- 10458527 TI - Nitric oxide: a clinically important amniotic fluid marker to distinguish between intra-amniotic mycoplasma and non-mycoplasma infections. AB - The objective of this study was to determine whether the measurements of amniotic fluid nitric oxide metabolite (NOx: nitrate + nitrite) concentrations could be a clinically useful marker to differentiate between intra-amniotic mycoplasma and nonmycoplasma infections. Amniocentesis was performed on 76 pregnant women with suspicion of intra-amniotic infection. Intra-amniotic infection was defined as the presence of a positive amniotic fluid culture with either mycoplasma or nonmycoplasma infections. Rapid amniotic fluid tests for Gram stain, glucose, leukocyte counts, interleukin-6, and NOx were performed. Amniotic fluid NOx was measured with aspergillus nitrate reductase and Griess reagent. Interleukin-6 was determined by enzyme immunoassays. Amniotic fluid NOx and interleukin-6 were normalized by amniotic fluid creatinine levels. Patients with intra-amniotic mycoplasma (n = 7) and nonmycoplasma infections (n = 8) had significantly higher amniotic fluid leukocyte counts and interleukin-6 concentrations and significantly lower amniotic fluid glucose levels than noninfected controls (n = 61). Amniotic fluid concentrations of NOx were significantly higher in those with intraamniotic nonmycoplasma infection as compared to those with intraamniotic mycoplasma infection and noninfected controls (NOx: 3.35+/-0.74 vs. 2.03+/-0.41 micromol/mg creatinine, p = 0.005, and 3.35+/-0.74 vs. 1.72+/-0.07 micromol/mg creatinine, p < 0.0001, respectively). However, patients with intra-amniotic mycoplasma infection did not differ significantly from noninfected controls. Our data indicate that clinical characteristics of intra-amniotic mycoplasma infection may differ from intra-amniotic nonmycoplasma infection. As delivery is not always indicated in intra-amniotic mycoplasma infection, elevated rapid amniotic fluid tests (leukocyte counts, interleukin-6, and glucose) may not be appropriate in the clinical management of intra-amniotic mycoplasma infection. In addition to these rapid amniotic fluid tests, incorporation of the measurement of amniotic fluid NOx may be of clinical importance in the differentiation and management of patients with suspected intra-amniotic mycoplasma and nonmycoplasma infection. PMID- 10458528 TI - Accuracy of strategies for monitoring fetal heart rate in labor. AB - We investigated whether the accuracy of auscultation could be improved with the use of a heart rate meter. Six fetal heart rate (FHR) traces were presented in a random sequence to 30 subjects using a customized computer program in each of three modalities: auscultation by counting alone, auscultation with the aid of an FHR meter, and visual assessment. The following characteristics were assessed: baseline rate, baseline variability, periodic change, and interpretation of the trace. For baseline rate, counting was associated with consistent underestimation of the FHR, which became more evident as the heart rate increased. The variation observed with each method was greatest with counting. For baseline variability, the proportion of correct responses using a meter was comparable to visual assessment, whereas counting was significantly less effective in 4 of 6 traces. For periodic change, the use of a meter was superior to counting in 4 of 6 traces, but both were inferior to visual assessment in 4 of 6 traces. In the interpretation of the trace, the use of a meter was again superior to counting, but both were inferior to visual assessment. Discrepancies were most marked in the least reassuring traces. Assessment of the FHR is significantly more accurate with the aid of a heart rate meter, and reduces interobserver variation. PMID- 10458529 TI - Hemoglobinopathy screening in pregnancy: comparison of two protocols. AB - This study was designed to determine the ability of a hemoglobinopathy screening protocol involving sickle solubility testing and red blood cell (RBC) indices to identify at-risk pregnancies. Retrospective chart review of all patients registering for prenatal care at the New York Hospital/Cornell Medical Center prenatal clinic in 1996 was the study design. All patients had RBC indices as well as hemoglobin electrophoresis. RBC indices of those with normal and abnormal electrophoreses were compared. Comparison of protocols involving universal hemoglobin electrophoresis and selective use of hemoglobin electrophoresis were compared. Student's t-test was used for statistical analysis. There were 36 carriers of hemoglobinopathy traits in 631 patients screened (5.7%). Four (three with hemoglobin C trait and one with hemoglobin D trait) had normal RBC indices and presumably would have had negative sickle solubility testing. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of a protocol with selective use of hemoglobin electrophoresis would have been 88.9, 79.4, 20.8, and 99.2%, respectively. Cost analysis reveals a difference of $11,384 or $18/patient less in a protocol with selective use of hemoglobin electrophoresis. Although a protocol involving solubility testing with RBC indices will not identify every carrier of a hemoglobinopathy trait, it may be appropriate in some populations. PMID- 10458530 TI - Second-trimester abortion caused by Capnocytophaga sputigena: case report. AB - Intra-amniotic infection is often the cause of a second-trimester abortion. The bacterial species involved include bacteria with low pathogenicity like Ureaplasma urealyticum and various Mycoplasma species. In this case we describe an intra-amniotic infection caused by Capnocytophaga sputigena, often found in the normal bacterial flora of the oral cavity, but not in the vagina. Oral sex during pregnancy was the most probable source of the infection. The aborted fetus showed signs of pneumonia upon histologic examination. The bacterial species was identified using broad-spectrum 16S rDNA polymerase chain reaction (PCR) directly from the amniotic fluid and after bacterial culture. Amniotic fluid glucose was below detection level, confirming the presence of an intra-amniotic infection. PMID- 10458531 TI - Fatigue in pregnancy: a comparative study. AB - The objective of this study was to use validated scales to determine if fatigue is increased during the first trimester of pregnancy compared to nonpregnant women with similar demographic characteristics. Women between 6 and 12 weeks' gestation were invited to complete a demographics questionnaire and the Numerical Rating Scale for Fatigue (NRS-F), State Trait Anxiety Inventory (STAI), and Beck Depression Index (BDI-II). Nonpregnant women were also invited to participate. Statistical analysis was performed using Mann-Whitney U, Fisher's exact, Spearman correlation, and Monte Carlo tests where appropriate; significance was assumed at p <0.05. Twenty pregnant and 15 nonpregnant patients were enrolled. There was no difference in age, parity, BMI, race, marital status, education, income or hours worked outside the home between pregnant and nonpregnant subjects. Pregnant women reported greater number of hours spent sleeping each day (8 [7-10] vs. 7 [6-9], p = 0.03). There was no significant difference in BDI-II, STAI-S, or STAI-T scores, but pregnant women had significantly higher scores on the NRS-F test (72.5 [20 88] vs. 36 [18-94], p <0.05). Women in the first trimester of pregnancy experience significantly greater fatigue compared to a similar group of nonpregnant women. PMID- 10458532 TI - Pregnancy and renal tubular acidosis. AB - Renal tubular acidosis (RTA) is uncommonly encountered in pregnancy. The risk for these women to develop pregnancy-induced hypertension has not been previously described. The renal defect noted in these women, aggravated by the normal hypervolemia of pregnancy, may predispose to hypertension. Three pregnancies in two women with RTA type 1 developed persistent diastolic hypertension in the third trimester. Mild renal insufficiency was noted in each woman as defined by serum creatinine of 0.9-1.1 and 1.4-1.6 mg/dL, respectively. Vaginal delivery was achieved in each without complications. Blood pressures returned to normal following each pregnancy. Pregnancy-induced hypertension developed in each of three pregnancies in two patients with RTA type 1. The risk for these women to develop pregnancy-induced hypertension may be associated with the higher reported risk in women with underlying renal disease. PMID- 10458533 TI - Management of severe neonatal anemia due to fetomaternal transfusion. AB - Three cases of severe neonatal anemia due to fetomaternal transfusion are reported. The key features that lead to early diagnosis were the maternal history, fetal monitoring, the clinical and laboratory findings of anemia, and a negative Coombs test. Diagnosis was confirmed by a rapid Kleihauer-Betke test. A partial exchange transfusion was performed in two of the three neonates with rapid clinical and hematological improvement. As two patients showed signs of heart decompensation, a partial exchange transfusion was performed with good success. In patients presenting with severe subacute or chronic anemia and heart failure, a partial exchange transfusion may be preferable to that of simple transfusion associated with diuretics. PMID- 10458534 TI - The incidence and severity of shoulder dystocia correlates with a sonographic measurement of asymmetry in patients with diabetes. AB - The objective of this paper is to examine the relationship between fetal asymmetry measured sonographically and the incidence and severity of shoulder dystocia in diabetic patients. Ultrasound data were collected retrospectively from examinations of women with gestational and pregastational diabetes who delivered at University of California, Irvine Medical Center from 1993-1995. Sonographic fetal asymmetry was quantified by calculating the difference between the abdominal diameter and the biparietal diameter in centimeters (AD-BPD). The residual AD-BPD was a patient's actual AD-BPD at the time of the ultrasound minus the mean AD-BPD obtained in our population at the patient's gestational age. The correlations between fetal asymmetry and the incidence and severity of shoulder dystocia were assessed using an analysis of variance as well as a logistic regression analysis. Mild shoulder dystocia was defined as a delivery requiring McRobert's maneuver and/or suprapubic pressure, while severe shoulder dystocia was assessed when delivery of the posterior arm with Wood's corkscrew maneuver was required. One hundred twenty-three women met the inclusion criteria for the study. Dividing the cohort into three groups based on AD-BPD residual values resulted in the following AD-BPD residual ranges and incidences of shoulder dystocia: Group I, -1.80 to -0.32 cm (9.8%), Group II, -0.31 to 0.32 cm (19.5%), and Group III .33 to 2.0 cm (34.1%), (p <0.03). The residual AD-BPD difference correlated with the incidence of shoulder dystocia after controlling for maternal age, weight, parity, birth weight, and the gestational age at ultrasound (P <0.03). Similar results were found with regards to dystocia severity as the mean residual AD-BPD difference between those with no dystocia, mild dystocia, and severe shoulder dystocia was -0.09, 0.23, and 0.46 cm, respectively, (p <0.006). The residual AD-BPD correlated with the severity of shoulder dystocia after controlling for the above-mentioned confounding variables (p <0.05) in a regression analysis. There is a direct correlation in diabetic patients between the level of fetal truncal asymmetry measured sonographically and the incidence and severity of shoulder dystocia. PMID- 10458535 TI - Control of infectious diseases. AB - Deaths from infectious diseases have declined markedly in the United States during the 20th century. This decline contributed to a sharp drop in infant and child mortality and to the 29.2-year increase in life expectancy. In 1900, 30.4% of all deaths occurred among children aged <5 years; in 1997, that percentage was only 1.4%. In 1900, the three leading causes of death were pneumonia, tuberculosis (TB), and diarrhea and enteritis, which (together with diphtheria) caused one third of all deaths. Of these deaths, 40% were among children aged <5 years. In 1997, heart disease and cancers accounted for 54.7% of all deaths, with 4.5% attributable to pneumonia, influenza, and human immunodeficiency virus (HIV) infection. Despite this overall progress, one of the most devastating epidemics in human history occurred during the 20th century: the 1918 influenza pandemic that resulted in 20 million deaths, including 500,000 in the United States, in <1 year-more than have died in as short a time during any war or famine in the world. HIV infection, first recognized in 1981, has caused a pandemic that is still in progress, affecting 33 million people and causing an estimated 13.9 million deaths. These episodes illustrate the volatility of infectious disease death rates and the unpredictability of disease emergence. PMID- 10458536 TI - Meningococcal disease--New England, 1993-1998. AB - Neisseria meningitidis, a leading cause of bacterial meningitis and sepsis in children and young adults in the United States, causes both sporadic disease and outbreaks. Preventing and controlling meningococcal disease remains a public health challenge because of the multiple serogroups and the limitations of available vaccines. Vaccination with the polysaccharide meningococcal vaccine, which protects against serogroups A, C, Y, and W135 of N. meningitidis, is recommended by the Advisory Committee on Immunization Practices (ACIP) for controlling outbreaks but routine vaccination is not recommended for control of sporadic cases. During 1998, a cluster of meningococcal disease cases occurred in Rhode Island, and although the situation did not meet ACIP criteria for an outbreak, the Rhode Island Department of Health recommended vaccination of all residents aged 2-22 years. This action stimulated controversy in Rhode Island and the rest of New England (Connecticut, Maine, Massachusetts, New Hampshire, and Vermont) and prompted a review of the epidemiology of meningococcal disease in the region. This report describes meningococcal disease data reported to the region's state health departments during 1993-1998 and discusses the situation in Rhode Island. PMID- 10458537 TI - Progress toward poliomyelitis eradication during armed conflict--Somalia and southern Sudan, January 1998-June 1999. AB - In 1988, the Regional Committee of the World Health Organization (WHO) for the Eastern Mediterranean Region adopted a resolution to eliminate poliomyelitis from the region by 2000. Somalia and parts of southern Sudan have persons living in areas where there is ongoing armed conflict and poor infrastructure (e.g., health care facilities, schools, roads, and power plants). Under these conditions, conducting National Immunization Days (NIDs) and acute flaccid paralysis (AFP) surveillance is difficult. This report summarizes NIDs in Somalia during 1997 and 1998 and in southern Sudan during 1998 and 1999 and establishment of AFP surveillance in northern Somalia and southern Sudan. PMID- 10458538 TI - Beta-estradiol-induced decrease in IL-12 and TNF-alpha expression suppresses macrophage functions in the course of Listeria monocytogenes infection in mice. AB - Mice treated with a contraceptive dose of beta-estradiol (E2) demonstrated changes in their macrophage (Mphi) number and functions. While E2 increased and decreased the Mphi number in PBMC and PEC respectively, it enhanced the in vitro phagocytosis of FITC-labeled beads by both cells. E2 treatment also enhanced the phagocytic function of Mphi as assessed by the in vivo carbon clearance assay. In contrast, the in vitro intracellular killing function of adherent cells in peritoneal exudate cells (PEC) against Listeria monocytogenes decreased after E2 treatment. In line with the decrease in the intracellular killing function, the E2-treated mice showed an impaired protection against L. monocytogenes infection. To clarify the mechanism of the E2-mediated suppression of the protective response against L. monocytogenes infection, we next analyzed the cytokine expression by PEC in E2-treated L. monocytogenes-infected mice. On day 5 of the infection, the expression of IL-12, TNF-alpha and IL-10 by adherent PEC from the E2-treated mice was lower than that from the control-infected mice. The decrease in the cytokine expression by adherent PEC of E2-treated mice coincided with the decrease of IFN-gamma expression, and the increase in the IL-4, IL-10 and TGF beta expressions by non-adherent PEC. These results revealed two aspects of the effects of E2 on Mphi. Even though E2 was found to enhance Mphi phagocytosis, the anti-bacterial function was suppressed. This suppression may be mediated by the inhibition of both IL-12 and TNF-alpha which play important roles in the protective response against intracellular bacteria. PMID- 10458539 TI - Effects of Chlorella vulgaris on bone marrow progenitor cells of mice infected with Listeria monocytogenes. AB - In this study we investigated the effects of the treatment with Chlorella vulgaris extract (CVE) on the hematopoietic response of granulocyte-macrophage colony-forming unit (CFU-GM) of mice infected with a sublethal dose of Listeria monocytogenes (1 x 10(4) organisms/animal). CVE was given orally as 50 mg/kg/day for 5 days. In the CVE treated/infected groups L. monocytogenes was administered at the end of CVE treatment. The colony stimulating activity of the serum (CSA) was also studied in all groups. Although no effects on CFU-GM, as compared to controls, were observed in the groups receiving CVE alone, the extract produced an increase in CSA levels as compared to controls. On the other hand, the presence of the infection led to a significant reduction in the numbers of CFU-GM as observed at 48 and 72 h after the infection, in spite of the significant increase in serum CSA activity. CVE treatment of infected animals restored the numbers of CFU-GM to control levels. In the treated/ infected group the increased serum CSA was significantly higher than that observed in the only infected group. The CVE treatment (50 and 500 mg/kg) of mice infected with a dose of 3 x 10(5) bacteria/animal, which was lethal for all the non-treated controls, produced a dose-response protection which led to a 20 and 52% survival, respectively. These results demonstrated that CVE produces a significant increase in the resistance of the animals infected with L. monocytogenes, and that this protection is due, at least in part, to increased CFU-GM in the bone marrow of infected animals. PMID- 10458540 TI - Impaired T and NK cell response of bone marrow and peripheral blood stem cell products to interleukin (IL)-2. AB - The function of steady-state and interleukin (IL)-2-co-cultured mononuclear cells differs significantly between bone marrow (BM) products, growth factor-mobilized peripheral blood stem cell (PSC) products and normal peripheral blood mononuclear cells (PBMC). The natural killer (NK) cell activity and T cell proliferative response of PSC products from non-Hodgkin's lymphoma (NHL) patients are significantly higher than that of BM products and similar to normal PBMC. However, following a five-day co-culture with IL-2 (100 IU/ml), the NK activity of PSC, PBMC, and BM products (lytic units) was increased 176-, 40-, and 14-fold, respectively, compared to that observed prior to IL-2 culture. In contrast, lymphokine activated killer (LAK) cytotoxicity prior to IL-2 culture was low in PSC and BM products and normal PBMC, but was significantly increased in PSC products and PBMC following IL-2 co-culture. The proliferative response of PSC and BM products to the T cell mitogen phytohemagglutinin (PHA) was significantly lower than that observed with normal PBMC; however, PSC had a significantly higher response than cells from BM products. Similar patterns of T cell PHA mitogenic response were observed after IL-2 co-culture. In addition, the IL-2 mitogenic responses of IL-2-co-cultured PSC and BM products were also significantly lower than that observed with PBMC co-cultured with IL-2. The IL-2 mitogenic response of PBMC was also significantly increased compared to prior to IL-2 co-culture; whereas, the IL-2 mitogenic responses from PSC and BM cells were not. In summary, co-culture with IL-2 can increase the NK and LAK cell cytotoxicity of PSC and BM products from NHL patients, but IL-2 co-culture does not improve T cell function within either BM or PSC products. PMID- 10458541 TI - Effects of salmeterol on host resistance to Trichinella spiralis in rats. AB - Salmeterol is a long-acting beta2-adrenoreceptor agonist. The compound has previously been screened for immunotoxic potential in a repeated dose toxicity study in rats for 28 days. The total serum IgG levels were increased at dose levels of 2 and 10 mg/kg/day. Presently, salmeterol was studied in an immune function assay addressing the host resistance to Trichinella spiralis parasites. Rats were daily treated with salmeterol for 28 days at dose levels of 0, 2, 6 and 10 mg/kg/day. On day 29, the animals were infected with T. spiralis parasites. After six weeks, host resistance was examined. The numbers of T. spiralis muscle larvae in the tongue nor the inflammatory reactions around the encapsulated larvae were affected by salmeterol treatment. The yield of muscle larvae in the whole carcass was not changed either. The IgM, IgA and IgE antibody responses to T. spiralis were unaffected. Only at the highest dose level tested, the anti-T. spiralis IgG antibody response was decreased significantly. However, salmeterol's interference with the generation of anti-T. spiralis antibodies of the IgG subclass apparently did not adversely affect the resistance to infection. PMID- 10458542 TI - The preoperative administration of lentinan ameliorated the impairment of natural killer activity after cardiopulmonary bypass. AB - The aim of this study was to determine whether the preoperative administration of lentinan, which is used clinically to activate T cell function in cancer patients, prevents the impairment of lymphocyte function after cardiopulmonary bypass (CPB). A total of 25 adults undergoing coronary artery bypass grafting were enrolled in this study. Lentinan (2 mg) was given to 10 randomly selected patients 7 d before surgery, while the other 15 patients were considered as a control. The white blood cell count, percentage of lymphocytes, subsets of lymphocytes, and natural killer cell activity were measured preoperatively, immediately after CPB and 1, 3, and 6 d after surgery. The white blood cell counts and the percentage of lymphocytes were not significantly different between the two groups; however, the percentage of CD4-positive cells in the lentinan group recovered to normal more rapidly than in the control group. Although natural killer cell activity was impaired in the control group after CPB, it maintained a nearly normal level in the lentinan group. The preoperative administration of lentinan for patients undergoing CPB ameliorated the impairment of natural killer activity and promoted the rapid recovery of CD4-positive cells. PMID- 10458543 TI - Sonographic diagnosis of intrauterine growth restriction (IUGR) by fetal transverse cerebellar diameter (TCD)/abdominal circumference (AC) ratio. AB - OBJECTIVE: To evaluate the validity of TCD/AC ratio in predicting IUGR. STUDY DESIGN: Prospective descriptive analysis. SETTING: Maharaj Nakorn Chiang Mai Hospital, Chiang Mai University. SUBJECTS: Women meeting the inclusion criteria consisting of: (1) singleton pregnancies after 28 weeks of gestation; (2) known accurate gestational age; and (3) clinically suspected IUGR. METHODS: The patients were sonographically examined for TCD/AC ratio. The best cut-off value of TCD/AC ratio in predicting IUGR was determined by a receiver operating characteristic (ROC) curve. The fetus with a TCD/AC ratio greater than the cut off value would be antenatally diagnosed as IUGR for every gestational week. Standard definition of IUGR was a low birthweight, less than the 10th percentile. RESULTS: One hundred and sixty-seven pregnancies with suspected IUGR were analyzed. The prevalence of IUGR among the study group was 51.5%. The best cut off value of the TCD/AC ratio for predicting IUGR was 15.4%, giving the sensitivity, specificity, positive predictive value and negative predictive value of 73.26%, 80.25%, 79.75%, and 73.86%, respectively. CONCLUSION: The sonographic fetal TCD/AC ratio as a gestational age-independent method can be helpful in antenatal diagnosis of IUGR, especially in pregnancy with uncertain gestational age. PMID- 10458545 TI - Tumor markers in endometriosis. AB - OBJECTIVES: The objective of the present study was to determine the concentrations of CA 125, CA 15-3, CA 19-9, carcioembryogenic antigen (CEA), alpha-fetoprotein (AFP) and beta-2 microglobulin (B2MG) in patients with pelvic endometriosis. METHOD: Fifty women were divided into two groups: group A (control) had no endometriosis or other diseases, and group B consisted of 35 women with pelvic endometriosis. All women were submitted to serum determination of CA 125, CA 15-3, CA 19-9, CEA, AFP and B2MG. Samples were collected during the menstrual cycle and 1 week later. RESULTS: Mean CA 125 concentrations were altered in patients with endometriosis, but all 50 patients studied presented normal CEA, AFP and B2MG concentrations. Small variations detected in CA 19-9 and CA 15-3 had no statistical significance. CONCLUSION: CA 125 is the only important marker in the diagnosis of stages III/IV of endometriosis, especially when blood samples for its determination are obtained during the first 3 days of the menstrual cycle. PMID- 10458544 TI - Cervical ectopy and the transformation zone measured by computerized planimetry in adolescents. AB - OBJECTIVE: To determine whether cervical ectopy and the transformation (T) zone were larger in adolescents using oral contraceptives (OCs) compared to depot medroxyprogesterone acetate (DMPA). METHOD: Cervical photographs were taken on 91 adolescents in Baltimore, Maryland. Ectopy and T zone size were measured using computerized planimetry. Patients provided reproductive and sexual history. Correlates of ectopy and T zone size were identified by proportional odds and linear regression models, respectively. RESULT: Twenty-five women had no ectopy. Parity was independently associated with greater ectopy (OR 3.9, 95% CI 1.37 11.11). Predictors of smaller ectopy were douching (OR 0.23, 95% CI 0.09-0.65), and shorter sexual history (OR 0.20, 95% CI 0.05-0.74). Neither OC or DMPA were associated with ectopy. Predictors of greater T zone size were years since menarche (13.8 mm2/year, P = 0.003) and OCs (54.7 mm2, P = 0.05). DMPA predicted a smaller T zone (-67.0 mm2, P = 0.01). CONCLUSION: Computerized planimetry provides standardized measurements. Douching and sexual activity may decrease ectopy through increased squamous metaplasia. Hormonal contraceptives were not associated with ectopy in adolescents. However, long-term progestin use may decrease T zone size. PMID- 10458546 TI - The prevalence of domestic violence against pregnant women in a Chinese community. AB - OBJECTIVE: To study the incidence of domestic violence in pregnant women attending the antenatal clinic of a local teaching hospital. STUDY DESIGN: All pregnant women attending their first antenatal clinic in Tsan Yuk Hospital between 11th August and 3rd November, 1998 were interviewed by a designated research nurse (Y.Y.J.L.) using a standard questionnaire (Abuse Assessment Screen) to detect the incidence of domestic violence, the nature of violence, the frequency of violence and the perpetrator of abuse. Demographic factors of the abused group were compared with those of the non-abused group using student's t test and chi-square test. RESULTS: Pregnant women (631) were interviewed; 113 of them (17.9%) had a history of abuse; 99 women (15.7%) had been abused in the last year; 27 of them (4.3%) had been abused during their current pregnancy; 59 women (9.4%) had been sexually abused in the last year. The husband was the perpetrator in the majority of cases. The nature of violence during pregnancy was mainly psychological in the form of threats of abuse without any physical injury. Risk factors included unplanned pregnancy (P = 0.002) and women with husbands/partners who were unemployed or manual workers (P < 0.05). Unexpectedly, domestic violence occurred more commonly in permanent local residents rather than new immigrants (P < 0.05). CONCLUSION: This is probably the first study on the incidence of domestic violence in pregnant women in a Chinese community. The incidence is comparable to that from American studies. Routine screening with structured questions during the antenatal visits is necessary in order to identify the abused women so as to prevent potential trauma and to interrupt existing abuse. PMID- 10458547 TI - Knowledge and practice of emergency contraception among Nigerian youths. PMID- 10458548 TI - Rubella immunity in pregnant Turkish women. PMID- 10458549 TI - Antacids vs. antacids plus non-prescription ranitidine for heartburn during pregnancy. PMID- 10458550 TI - Amniotic fluid and maternal characteristics in Chinese pregnancies dated by early ultrasound biometry. PMID- 10458551 TI - Diagnosis and position shift between the first and the second fetus in a diamniotic twin pregnancy. PMID- 10458552 TI - Leiomyomatosis peritonealis disseminata with malignant transformation. PMID- 10458553 TI - Laparoscopic loop ligatures for bladder repair during laparoscopic surgery. AB - In this paper, we report a case where a bladder perforation occurred during a laparoscopically assisted vaginal hysterectomy and was repaired by laparoscopic loop ligatures. This is the first case report of using the laparoscopic loop ligatures to close the bladder perforations. The loop ligature is an easy and quick procedure, which can be performed by most surgeons who take the time to learn the endoscopic suturing techniques. PMID- 10458554 TI - A pictorial chart for managing common menstrual disorders in Nigerian adolescents. PMID- 10458555 TI - Some ethical and legal issues in assisted reproductive technology. AB - The potential and actual applications of reproductive technologies have been reviewed by many governmental committees, and laws have been enacted in several countries to accommodate, limit and regulate their use. Regulatory systems have nevertheless left some legal and ethical issues unresolved, and have caused other issues to arise. Issues that regulatory systems leave unresolved, or that systems have created, include disposal of embryos that remain after patients' treatments are concluded, and multiple implantation and pregnancy. This may result in risks to maternal, embryonic and neonatal life and health, and the contentious relief that may be achieved by selective reduction of multiple pregnancies. A further concern arises when clinics must (or choose to) publicize their success rates, and they compete for favorable statistics by questionable patient selection criteria and treatment priorities. PMID- 10458556 TI - ACOG practice bulletin. Prevention of Rh D alloimmunization. Number 4, May 1999 (replaces educational bulletin Number 147, October 1990). Clinical management guidelines for obstetrician-gynecologists. American College of Obstetrics and Gynecology. AB - The reduction in the incidence of Rh D alloimmunization is a prototype for the effectiveness of preventive medicine. Some controversies remain, however, such as the use of anti-D immune globulin in patients with either threatened abortion or antenatal hemorrhage. Similarly, it may not be cost-effective either to screen all Rh D-negative patients with an indirect Coombs test at 24-28 weeks of gestation or to screen all postpartum patients for excessive fetomaternal hemorrhage. PMID- 10458557 TI - Surface characterization and platelet adhesion studies of plasma polymerized phosphite and its copolymers with dimethylsulfate. AB - Due to its capability for using a wide variety of organic and inorganic compounds as the monomer source, including those which are not polymerizable by conventional methods, plasma polymerization has been used to incorporate various functional groups onto the substrates of interest. Plasma polymerization of trimethylphosphite and triisopropylphosphite was attempted with an aim to deposit a thin film with phosphorous-containing functional groups such as phosphate or phosphite onto the glass substrate. Sulfur-containing functional groups, such as sulfonate, were incorporated as well with the addition of dimethylsulfate to the phosphite monomer inlet. These plasma polymers and plasma copolymers were very hydrophilic. Incomplete surface coverage was noticed under the processing conditions of low phosphite flow rate, low RF input power and shorter deposition time. Extended deposition duration and higher power input resulted in the formation of voids and pits on the plasma polymer surface. In addition, the surface coverage of the plasma copolymers becomes discontinuous if the flow rate ratio of dimethylsulfate to triisopropylphosphite is too high. In vitro platelet adhesion studies indicated the plasma polymers and plasma copolymers with continuous surface coverage are less platelet activating than the glass control. Moreover, the addition of dimethylsulfate into the phosphite monomer flow led to a further reduction in platelet adhesion and activation than the comparable one. This may be attributed to the effect of the sulfur-containing functionalities, such as sulfonate or sulfate, found on the plasma copolymers. PMID- 10458558 TI - Radiation and chemical crosslinking promote strain hardening behavior and molecular alignment in ultra high molecular weight polyethylene during multi axial loading conditions. AB - The mechanical behavior and evolution of crystalline morphology during large deformation of eight types of virgin and crosslinked ultra high molecular weight polyethylene (UHMWPE) were studied using the small punch test and transmission electron microscopy (TEM). We investigated the hypothesis that both radiation and chemical crosslinking hinder molecular mobility at large deformations, and hence promote strain hardening and molecular alignment during the multiaxial loading of the small punch test. Chemical crosslinking of UHMWPE was performed using 0.25% dicumyl peroxide (GHR 8110, GUR 1020 and 1050), and radiation crosslinking was performed using 150 kGy of electron beam radiation (GUR 1150). Crosslinking increased the ultimate load at failure and decreased the ultimate displacement of the polyethylenes during the small punch test. Crosslinking also increased the near-ultimate hardening behavior of the polyethylenes. Transmission electron microscopy was used to characterize the crystalline morphology of the bulk material, undeformed regions of the small punch test specimens, and deformed regions of the specimens oriented perpendicular and parallel to the punch direction. In contrast with the virgin polyethylenes, which showed only subtle evidence of lamellar alignment, the crosslinked polyethylenes exhibited enhanced crystalline lamellae orientation after the small punch test, predominantly in the direction parallel to the punch direction or deformation axis. Thus, the results of this study support the hypothesis that crosslinking promotes strain hardening during multiaxial loading because of increased resistance to molecular mobility at large deformations effected by molecular alignment. The data also illustrate the sensitivity of large deformation mechanical behavior and crystalline morphology to the method of crosslinking and resin of polyethylene. PMID- 10458559 TI - Unified wear model for highly crosslinked ultra-high molecular weight polyethylenes (UHMWPE). AB - Crosslinking has been shown to improve the wear resistance of ultra-high molecular weight polyethylene in both in vitro and clinical in vivo studies. The molecular mechanisms and material properties that are responsible for this marked improvement in wear resistance are still not well understood. In fact, following crosslinking a number of mechanical properties of UHMWPE are decreased including toughness, modulus, ultimate tensile strength, yield strength, and hardness. In general, these changes would be expected to constitute a precursor for lower wear resistance, presenting a paradox in that wear resistance increases with crosslinking. In order to understand better and to analyze this paradoxical behaviour of crosslinked UHMWPE, we investigated the wear behavior of (i) radiation-crosslinked GUR 1050 resin, (ii) peroxide-crosslinked GUR 1050 resin and (iii) peroxide-crosslinked Himont 1900 resin using a bi-directional pin-on disk (POD) machine. Wear behavior was analyzed as a function of crystallinity, ultimate tensile strength (UTS), yield strength (YS), and molecular weight between crosslinks (Mc). The crosslink density increased with increasing radiation dose level and initial peroxide content. The UTS, YS, and crystallinity decreased with increasing crosslink density. While these variations followed the same trend, the absolute changes as a function of crosslink density were different for the three types of crosslinked UHMWPE studied. There was no unified correlation for the wear behavior of the three types of crosslinked UHMWPE with the crystallinity, UTS and YS. However, the POD wear rate showed the identical linear dependence on Mc with all three types of crosslinked UHMWPEs studied. Therefore, we have strong evidence to propose that Mc or crosslink density is a fundamental material property that governs the lubricated adhesive and abrasive wear mechanisms of crosslinked UHMWPEs, overriding the possible effects of other material properties such as UTS, YS and crystallinity on the wear behavior. PMID- 10458560 TI - Wear-testing of a temporomandibular joint prosthesis: UHMWPE and PTFE against a metal ball, in water and in serum. AB - For a temporomandibular joint prosthesis, an estimation of the wear rate was needed, prior to patient application. Therefore, we determined the in vitro wear rate of the ball-socket articulation of this prosthesis, consisting of a metal head and an ultra-high molecular weight polyethylene (UHMWPE) cup. The basic testing configuration consisted of one 8-mm diameter stainless-steel ball, rotating between two conforming cups with a minimum thickness of 5 mm. For validation of the testing apparatus, two cup materials, in two lubricants, were tested. Both cup materials, UHMWPE and polytetrafluoroethylene (PTFE) were tested in deionized water, as well as in a serum-based solution. For UHMWPE in serum, eight samples were tested, for the other combinations four samples. For UHMWPE, the tests ran for 7 million cycles, for PTFE between 0.8 and 1.7 million cycles. For UHMWPE, the wear rate was 0.006 and 0.47 (mm3/10(6) cycles), in water and in serum, respectively. For PTFE, the wear rate was 2.8 and 47 (mm3/10(6) cycles), in water and in serum, respectively. For reason that testing in serum simulates the in vivo situation best, it was concluded that the wear rate of the TMJ prosthesis articulation is 0.47 (mm3/10(6) cycles), which is considered acceptable. PMID- 10458561 TI - Properties of the poly(vinyl alcohol)/chitosan blend and its effect on the culture of fibroblast in vitro. AB - In this work, the properties of poly(vinyl alcohol) (PVA) and PVA/chitosan blended membranes were investigated by scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and electron spectroscopy for chemical analysis (ESCA). The SEM photographs show the PVA/chitosan blended membrane undergoes dramatic changes on the surface and bulk structure during the membrane formation. The DSC analysis shows that PVA and chitosan are not very compatible in the PVA/chitosan blended membrane, whereas the combination of two polymer chains of constitutionally different features is revealed. In addition, the surface of the PVA/chitosan blended membrane is enriched with nitrogen atoms at the ESCA analysis. These reflect the PVA membrane can be modified by blending with chitosan that in turn may affect the biocompatibility of the blended membrane. Therefore, adhesion and growth of fibroblasts on the PVA as well as PVA/chitosan blended membranes were investigated. Cell morphologies on the membranes were examined by SEM and cell viability was studied using MTT assay. It was observed that the PVA/chitosan blended membrane was more favorable for the cell culture than the pure PVA membrane. Cells cultured on the PVA/chitosan blended membrane had good spreading, cytoplasm webbing and flattening and were more compacting than on the pure PVA membrane. Consequently, the PVA/chitosan blended membrane may spatially mediate cellular response that can promote cell attachment and growth, indicating the PVA/chitosan blended membrane should be useful as a biomaterial for cell culture. PMID- 10458562 TI - Highly permeable polylactide-caprolactone nerve guides enhance peripheral nerve regeneration through long gaps. AB - We compared regeneration and functional reinnervation after sciatic nerve resection and tubulization repair with bioresorbable guides of poly(L-lactide-co epsilon-caprolactone) (PLC) and permanent guides of polysulfone (POS) with different degrees of permeability, leaving a 6 mm gap in different groups of mice. Functional reinnervation was assessed to determine recovery of motor, sensory and sweating functions in the hindpaw during four months postoperation. Highly permeable PLC guides allowed for faster and higher levels of reinnervation for the four functions tested than impermeable or low-permeable PLC guides, while semipermeable 30 and 100 kDa POS tubes yielded very low levels of reinnervation. The regeneration success rate was higher with PLC than with POS tubes. Morphometrical analysis of cross-sectional nerves under light microscopy showed the highest number of regenerated myelinated fibers at mid tube and distal nerve in high-permeable PLC guides. Impermeable PLC guides allowed slightly worse levels of regeneration, while low-permeable PLC guides promoted neuroma and limited distal regeneration. The lowest number of regenerated fibers were found in POS tubes. In summary, highly permeable bioresorbable PLC guides offer a suitable alternative for repairing long gaps in injured nerves, approaching the success of autologous nerve grafts. PMID- 10458563 TI - The reduced adsorption of lysozyme at the phosphorylcholine incorporated polymer/aqueous solution interface studied by spectroscopic ellipsometry. AB - Coating hydrogel polymers onto solid substrates can reduce the adsorption of proteins onto these surfaces, but the extent of the reduction in protein adsorption is strongly dependent on how the surface layer is coated. We have examined the effect of coating conditions on the structure of thin polymer films formed from a number of poly(methacrylate)-based hydrogel polymers via the dip coating method. We show in this work how the polarity of the solvent, the speed of lifting, and the annealing temperature affect the thickness and uniformity of ultrathin phosphorylcholine (PC)-incorporated polymer films coated on the surface of native oxide on silicon and the subsequent interaction of these coated surfaces with lysozyme molecules. Our results show that the uniformity of the polymer film, and thus the smoothness of the outer film surface, influence the extent of reduction in protein adsorption. We suggest that the reduction in lysozyme adsorption is the result of a layer of PC groups on the surface of the polymer film. The improvement of the smoothness of the film results in the formation of a close-packed PC layer on the outer surface of the polymer film, leaving few defects or cavities on which protein molecules can bind. PMID- 10458564 TI - A novel osteochondral implant. AB - A novel implant for the use as an osteochondral graft was designed. This implant was prepared by stepwise formation of calcium phosphate crystals within the matrix of a lyophilised collagen sponge. Chondrocytes were then grown on this material to create the osteochondral implant. The implant was characterized with light microscopy, scanning electron microscopy (SEM), electron diffraction crystallography (EDX), and IR. It was observed with IR that the implant had a peak, that was not found so distinctly in its components, at 1400 cm(-1), implying a strong interaction of the two main ingredients of the implant, calcium phosphate and collagen. This strong interaction was also shown in the graft degradation test while the untreated collagen sponge degraded rapidly (in one day) the mineral loaded implant was able to maintain its integrity for two weeks. In the chondrocyte culture medium degradation of the implant was shown by a decrease of the calcium content and calcium to phosphorous ratio. Also, EDX revealed the presence of sulfur one and two weeks after incubation, an element not found among the components of the implant, possibly due to the development of an extracellular matrix. SEM showed that the form of the crystals of calcium phosphate differed depending on whether they were prepared on the template, collagen, or in the absence of a template. The chondrocytes appeared to be growing in number on the implant and their shapes were morphologically normal. The chondrocyte loaded collagen-calcium phosphate composite could thus be considered a potential tissue engineered osteochondral implant. PMID- 10458565 TI - Platelet-derived microparticles on synthetic surfaces observed by atomic force microscopy and fluorescence microscopy. AB - Platelet activation on a thrombogenic surface includes the release of membrane derived microparticles that provide catalytic sites for blood coagulation factors. Here, we describe a quantitative investigation on the production and dimensions of platelet-derived microparticles observed on glass and polyethylene under aqueous conditions, using atomic force microscopy (AFM) and complementary fluorescence microscopy. The results show that contact-activated platelet microparticles are not evenly distributed over a thrombogenic surface, but in clusters in close proximity to adherent platelets. The microparticles are localized near the platelet periphery, and in some cases appear to emanate from platelet pseudopodia, suggesting that formation may result from vesiculation of the pseudopodia. The microparticles measured 125 +/- 21 nm (n = 73) in the x-y dimensions and 5.2 +/- 3.6 nm in height. The results compared closely with 125 +/ 22 nm width and 4.1 +/- 1.6 nm height obtained for control preparations of thrombin activated microparticles, that were filtered and deposited on glass. Large differences between the measured widths and heights of adsorbed microparticles suggest that platelet microparticles may undergo spreading after attachment to a surface. The adsorbed microparticles expressed platelet membrane receptor GPIIb/IIIa, and many expressed the platelet activation marker P-selectin as determined by fluorescence microscopy. The high number distribution of procoagulant microparticles per unit area of surface compared with platelets suggests that platelet-derived microparticles provide a mechanistic route for amplifying thrombus formation on a thrombogenic surface. PMID- 10458566 TI - Simultaneous superficial hyperthermia and external radiotherapy: report of thermal dosimetry and tolerance to treatment. AB - BACKGROUND: In vitro and animal studies indicate that a moderate temperature of 41 degrees C maintained for approximately 1 h will provide radiosensitization if radiation (RT) and hyperthermia (HT) are delivered simultaneously, but not with sequential treatment. A minimum tumour temperature of 41 degrees C is a more feasible goal than the goal of >42 degrees C needed for sequential treatment. METHODS AND MATERIALS: Forty-four patients with 47 recurrent superficial cancers received simultaneous external beam radiotherapy and superficial hyperthermia on successive IRB approved phase I/II studies. All lesions had failed previous therapy, 35 were previously irradiated (mean dose 52.7 Gy). Hyperthermia was delivered with 915 MHz microwave or 1-3.5 MHz ultrasound using commercially available applicators. The average dimensions of 19 lesions treated with microwave were 4.7 x 3.6 x 1.7 cm and the average dimensions of 28 lesions treated with ultrasound were 8.0 x 6.1 x 2.9 cm. The most common sites were chest wall (15 cases) and head and neck (21 cases). Temperatures were monitored at an average of six intratumoral locations using multisensor probes. The median number of hyperthermia treatments was three and the median radiation dose 30 Gy. Radiation dose per fraction was 4 Gy with hyperthermia and 2 Gy or 4 Gy (depending on protocol) on non-hyperthermia days. RESULTS: Six different measures of minimum monitored temperature and duration were found to be highly correlated with each other. There was nearly a one-to-one correspondence between minimum tumour time at or above 41 degrees C (Min t41) and minimum tumour Sapareto Dewey equivalent time at 42 degrees C (Min teq42). After four sessions 63% of cases had a per session average Sapareto Dewey equivalent time at 41 degrees C which exceeded 60 min in all monitored tumour locations. The complete and partial response rate in evaluable lesions were respectively 21/41 (51%) and 7/41 (17%) and were best correlated with site (chest wall showing best response). Toxicity consisted of 10/47 (21%) slow healing soft tissue ulcers which healed in all cases but required a median of 7 months. The most important predictors for chronic ulceration were cumulative radiation dose >80 Gy and complete response to treatment. CONCLUSIONS: Minimum tumour temperatures maintained for durations compatible in vitro with thermal radiosensitization (if RT and HT are delivered simultaneously) are clinically feasible and tolerable for broad but superficial lesions amenable to externally applied ultrasound or microwave hyperthermia. The current in-house protocol is evaluating the impact of more than four hyperthermia sessions on the overall thermal dose distribution and toxicity. PMID- 10458567 TI - Induction of vascular endothelial growth factor (VEGF) by hyperthermia and/or an angiogenesis inhibitor. AB - Intratumoral localization of vascular endothelial growth factor (VEGF) following administration of hyperthermia (HT) and/or anti-angiogenic drugs (TNP-470) was evaluated using SCC VII tumours in C3H/He mice. Hyperthermia at 44.0 degrees C for 30 min was given with a water bath on day 0. TNP-470 (100 mg/kg) was administered alone or after HT on day 0 and day 3. Histological changes on day 4 were evaluated by haematoxylin-eosin (HE) staining and immunohistochemical staining for VEGF. The percentage of the necrotic area relative to the entire tumour area (the % necrotic area) was measured on HE stains. The average % necrotic area of the untreated SCC VII tumours was 7%, while those of tumours treated with TNP-470 alone and HT alone were 27 or 65%, respectively. When HT and TNP-470 were combined, the % necrotic area was 82%, which was significantly higher than that caused by HT alone (p < 0.05). Immunohistochemical staining for VEGF in untreated SCC VII tumours was weak, although strong staining for VEGF was noted in untreated EMT-6 tumours of BALB/c mice, which have spontaneous central necrosis. After administration of HT and/or TNP-470, layer-shaped staining by VEGF was observed in the residual SCC VII tumour cells adjacent to the necrotic area. In conclusion, the expression of VEGF increased in response to administration of HT and/or TNP-470. Hypoxia caused by heat-induced vascular damage may be attributable to increased expression of VEGF in SCC VII tumours. PMID- 10458568 TI - Thermal radiosensitization of human tumour cell lines with different sensitivities to 41.1 degrees C. AB - While much work on radiosensitization by hyperthermia in the 43 degrees C and higher temperature range has been done, relatively little work has been done at temperatures in the 41-42 degrees C range. In this moderate hyperthermia range there are dramatic differences in the resistance of mammalian cells to hyperthermia. Therefore, thermal radiosensitization was measured in two human colon adenocarcinoma cell lines, one that expresses chronic thermotolerance and proliferates at 41.1 degrees C, NSY 42129 (NSY) cells and one that is slowly killed at 41.1 degrees C, HCT15 cells. Heat-resistant NSY cells were found to be more radioresistant than heat-sensitive HCT15 cells. Hyperthermia at 41.1 degrees C enhanced the radiation sensitivity in NSY cells, but no significant induction of heat-induced radiosensitization was observed in HCT15 cells. The radiation sensitivity induced by 41.1 degrees C in NSY cells appeared to be related to both intrinsic heat-induced radiosensitization (HIR) and cell-cycle redistribution at 41.1 degrees C. Incidentally, cells incubated at 41.1 degrees C for between 8-16 h displayed an identical radiosensitivity to those heated for 24 h. This result implies that modest hyperthermia for 2 h or more can have a radiosensitizing effect in heat-resistant cells. PMID- 10458569 TI - Electromagnetic thermal therapy power optimization for multiple source applicators. AB - The optimization of power deposition for electromagnetic (EM) thermal therapy is investigated. Several goal or objective functions are examined using a generalized mathematical formulation. These include maximization of: (1) target power absorption, (2) the ratio of target to non-target power absorption, (3) target power absorption weighted by the ratio of target to non-target power absorption, and (4) target power absorption subject to the constraint that the non-target high power volume ('hot spot' volume) is below a chosen level. The merit of these functions was retrospectively tested using an anatomic data base containing 38 cancer patients that were clinically heated with EM phased arrays. CT and/or MRI image data were used to define relevant anatomic geometries and tissue properties for finite element numerical models. Power optimization is achieved by variation of seven available control parameters (four amplitudes and three phases) for these clinical array devices. The results indicate that site dependent improvements in target power absorption can be achieved using these goal functions relative to a configuration that utilizes equal phase and amplitude for the sources. The relative merit among these various functions favours an optimization strategy that maximizes the target power absorption weighted by the ratio of target power to non-target power absorption. PMID- 10458570 TI - A HF EM installation allowing simultaneous whole body and deep local EM hyperthermia. AB - The structure and main features of a HF EM installation based upon a new approach for creating electromagnetic fields destined for whole body (WBH) and deep local (DLH) hyperthermia are discussed. The HF EM field, at a frequency of 13.56 MHz, is created by a coplanar capacity type applicator positioned under a distilled water filled bolus that the patient is lying on. The EM energy being released directly in the deep tissues ensures effective whole body heating to required therapeutic temperatures of up to 43.5 degrees C, whereas the skin temperature can be maintained as low as 39-40.5 degrees C. For DLH, the installation is equipped with additional applicators and a generator operating at a frequency of 40.68 MHz. High efficiency of the WBH applicator makes it possible to carry out the WBH procedure without any air-conditioning cabin. Due to this, a free access to the patient's body during the WBH treatment is provided and a simultaneous WBH/DLH or WBH/LH procedure by means of additional applicators is possible. Controllable power output in the range of 100-800 W at a frequency of 13.56 MHz and 50-350 W at a frequency of 40.68 MHz allows accurate temperature control during WBH, DLH and WBH/DLH procedures. SAR patterns created by the WBH and DLH applicators in a liquid muscle phantom and measured by means of a non-perturbing E-dipole are investigated. The scattered EM field strength measured in the vicinity of the operating installation during the WBH, DLH and WBH/DLH procedures does not exceed security standards. Examples of temperature versus time graphs in the course of WBH, DLH and WBH/DLH procedures in clinics are presented. The installation is successfully used in leading oncological institutions of Russia and Belarus, though combined WBH/DLH procedures are evidently more complicated and demand thorough planning and temperature measurements to avoid overheating. PMID- 10458571 TI - Technical note: Parasitic thermovoltage in the multithermocouple probe, its explanation and elimination. AB - The parasitic thermovoltage can influence the temperature measurement during hyperthermia using the miniature multithermocouple probes with a common wire. It was noticed that, when a thermocouple junction is placed in a sharp thermal gradient, a parasitic thermovoltage is added to the voltage of all remaining thermocouples situated in the direction to the tip of the probe. This article gives the theoretical explanation, experimental verification, and practical elimination of this phenomenon. PMID- 10458572 TI - Comparisons between the BSD-2000 Quad Amplifier and a new prototype solid state amplifier for deep regional hyperthermia. PMID- 10458573 TI - Bovine tuberculosis: the government reaffirms its strategy. PMID- 10458574 TI - Studies on the efficacy of intranasal vaccination for the prevention of experimentally induced parainfluenza type 3 virus pneumonia in calves. AB - The efficacy of intranasal vaccination in preventing or limiting disease of the lower respiratory tract induced by parainfluenza 3 (PI3) virus was evaluated under experimental conditions, using a commercially available live vaccine containing a temperature-sensitive strain of PI3 virus. In a preliminary study four colostrum-deprived calves were vaccinated intranasally at one week and again at two months of age, and two similar calves were given an intranasal placebo. After the second vaccination serum antibodies to PI3 virus were detected in all four vaccinated calves, but not in the control animals. Seventeen days after the second vaccination all six calves were challenged with virulent PI3 virus, and they were killed six days later. The clinical scores and the extent of pulmonary consolidation were reduced in the vaccinated animals; PI3 virus was detected in the upper and lower respiratory tract of the control calves but in none of the vaccinated calves. In a larger scale study with 14 colostrum-fed calves, seven were vaccinated at one week and again at five weeks of age, and seven were given an intranasal placebo. Two weeks after the second vaccination all 14 calves were challenged with virulent PI3 virus. The clinical scores and lung consolidation were significantly reduced in the vaccinated calves in comparison with the controls. Six days after infection, 10 of the 14 calves were killed; PI3 virus was detectable in the nasal secretions of all seven control calves but in only one of the vaccinated animals, and PI3 viral antigen was detected in the lungs of the control calves but not in those of the vaccinated animals. One of the vaccinated calves had developed a severe clinical response after the challenge, but it had only minor lung consolidation when killed. PMID- 10458575 TI - High carbon dioxide tension (PCO2) and the incidence of cardiac arrhythmias in rapidly growing broiler chickens. AB - The purpose of this investigation was to determine whether two-week-old rapidly growing broiler chickens with high metabolic activity have an increased risk of the development of heart failure three to five weeks later. The incidence of cardiac arrhythmias was assessed in broiler chickens with either a relatively high carbon dioxide tension (PCO2) or a low PCO2 in their venous blood. Their electrocardiograms (ECGS) were measured when the birds were between five and seven weeks old by means of a biotransplant which allowed them to move freely. Premature ventricular beats were observed in all the birds, but the largest numbers were observed in birds that had had a high PCO2 when they were two weeks old. PMID- 10458576 TI - Interdental acrylic stabilisation of canine tooth root and mandibular fractures in a dog. AB - A two-year-old labrador had bilateral open fractures of the apical region of the mandibular canine tooth roots and the adjacent mandible. The fractures were reduced and held in place with an interdental acrylic splint, after which the tooth roots and mandible healed directly. PMID- 10458577 TI - Practical evaluation of the fertilising capacity of frozen-thawed horse semen. PMID- 10458578 TI - Efficacy of two flubendazole formulations against Trichuris vulpis in naturally infected dogs. PMID- 10458579 TI - Body condition in end-of-lay hens: some implications. PMID- 10458581 TI - Equine viral arteritis in a gelding in the UK. PMID- 10458580 TI - The need for a veterinary antibiotic policy. AB - The international recognition of the 'stable to table' approach to food safety emphasises the need for appropriate and safe use of antibiotics in animal production. An appropriate use of antibiotics for food animals will preserve the long-term efficacy of existing antibiotics, support animal health and welfare and limit the risk of transfer of antibiotic resistance to humans. Furthermore, it may promote consumer confidence in the veterinary use of antibiotics. In advancing these arguments, the authors of this article argue that there is a need for a visible and operational policy for veterinary use of antibiotics, paying particular attention to the policies that are being developed in Denmark. PMID- 10458583 TI - Veterinary nursing examinations. PMID- 10458582 TI - Chemical destruction of horses. PMID- 10458584 TI - Watery milk. PMID- 10458585 TI - Fibrous epulides in a horse. PMID- 10458586 TI - Sedation of pot-bellied pigs. PMID- 10458587 TI - Funding for wildlife disease research. PMID- 10458588 TI - Richard Taylor memorial fund. PMID- 10458589 TI - Inflammatory mediators and stroke: new opportunities for novel therapeutics. AB - Contrary to previous dogmas, it is now well established that brain cells can produce cytokines and chemokines, and can express adhesion molecules that enable an in situ inflammatory reaction. The accumulation of neutrophils early after brain injury is believed to contribute to the degree of brain tissue loss. Support for this hypothesis has been drawn from many studies where neutrophil depletion blockade of endothelial-leukocyte interactions has been achieved by various techniques. The inflammation reaction is an attractive pharmacologic opportunity, considering its rapid initiation and progression over many hours after stroke and its contribution to evolution of tissue injury. While the expression of inflammatory cytokines that may contribute to ischemic injury has been repeatedly demonstrated, cytokines may also provide "neuroprotection" in certain conditions by promoting growth, repair, and ultimately, enhanced functional recovery. Significant additional basic work is required to understand the dynamic, complex, and time-dependent destructive and protective processes associated with inflammation mediators produced after brain injury. The realization that brain ischemia and trauma elicit robust inflammation in the brain provides fertile ground for discovery of novel therapeutic agents for stroke and neurotrauma. Inhibition of the mitogen-activated protein kinase (MAPK) cascade via cytokine suppressive anti-inflammatory drugs, which block p38 MAPK and hence the production of interleukin-1 and tumor necrosis factor-alpha, are most promising new opportunities. However, spatial and temporal considerations need to be exercised to elucidate the best opportunities for selective inhibitors for specific inflammatory mediators. PMID- 10458591 TI - Bolus injection versus slow infusion of [15O]water for positron emission tomography activation studies. AB - In positron emission tomography studies using bolus injection of [15O]water, activation responses reflect underlying CBF changes during a short time (15 to 20 seconds) after arrival of the bolus in the brain. This CBF sensitivity window may be too short for complex activation paradigms, however, particularly those of longer duration. To perform such paradigms, we used a slow infusion method of tracer administration to lengthen the CBF sensitivity window. The present study was designed to determine if this slow infusion technique yields similar results to a bolus injection with a short activation task involving memory for faces. When analyzed using statistical parametric mapping, scanning durations of either 90 or 120 seconds and a 90-second slow infusion schedule produced very similar results to a standard 60-second scan collected after bolus injection, indicating that statistically similar brain activation maps can be produced with the two infusion techniques. This slow infusion approach allows for increased flexibility in designing future studies in which a short CBF sensitivity window is a limiting factor. PMID- 10458590 TI - Reduction of cognitive and motor deficits after traumatic brain injury in mice deficient in poly(ADP-ribose) polymerase. AB - Poly(ADP-ribose) polymerase (PARP), or poly-(ADP-ribose) synthetase, is a nuclear enzyme that consumes NAD when activated by DNA damage. The role of PARP in the pathogenesis of traumatic brain injury (TBI) is unknown. Using a controlled cortical impact (CCI) model of TBI and mice deficient in PARP, the authors studied the effect of PARP on functional and histologic outcome after CCI using two protocols. In protocol 1, naive mice (n = 7 +/+, n = 6 -/-) were evaluated for motor and memory acquisition before CCI. Mice were then subjected to severe CCI and killed at 24 hours for immunohistochemical detection of nitrated tyrosine, an indicator of peroxynitrite formation. Motor and memory performance did not differ between naive PARP +/+ and -/- mice. Both groups showed nitrotyrosine staining in the contusion, suggest ing that peroxynitrite is produced in contused brain. In protoco 2, mice (PARP +/+, n = 8; PARP -/-, n = 10) subjected to CCI were tested for motor and memory function, and contusion volume was determined by image analysis. PARP -/- mice demonstrated improved motor and memory function after CC versus PARP +/+ mice (P < 0.05). However, contusion volume was not different between groups. The results suggest a detri mental effect of PARP on functional outcome after TBI. PMID- 10458592 TI - Quantitative magnetic resonance imaging in experimental hypercapnia: improvement in the relation between changes in brain R2 and the oxygen saturation of venous blood after correction for changes in cerebral blood volume. AB - Acute hypercapnia simultaneously induces increases in regional cerebral blood volume (rCBV) and the oxygen saturation of cerebral venous blood (Yv). Changes in both physiologic parameters may influence the changes in R2 (deltaR2) that can be measured in the brain with gradient echo magnetic resonance imaging. The authors examined the effect of incorporating independent measurements of the change in rCBV (deltarCBV) on the fidelity of the relation between deltaR2 and deltaYv in the setting of experimental hypercapnia. A two-dimensional T2-weighted gradient echo sequence was used to measure deltaR2 in the brain parenchyma of anesthetized rats in response to hypercapnia with respect to the control state. In parallel, estimates of rCBV were obtained using a three-dimensional steady-state approach in conjunction with a paramagnetic contrast agent during both control and hypercapnic states so that a deltarCBV could be calculated. Regional CBV values of 2.96 +/- 0.82% and 5.74 +/- 1.21% were obtained during the control and hypercapnic states, respectively, and linear relations between rCBV and CO2 tension in both arterial (r = 0.80) and jugular venous (r = 0.76) blood samples were obtained. When correlating deltaR2 directly with deltaYv, no clear relation was apparent, but a strong linear relation (r = 0.76) was observed when correction for deltarCBV was incorporated into the data analysis. These results are consistent with the current understanding of the mechanisms of blood oxygen level-dependent (BOLD) contrast and underscore the potential importance of taking into account deltarCBV when quantitative estimates of deltaYv from the "BOLD effect" are intended. PMID- 10458593 TI - Dynamic in vivo measurement of erythrocyte velocity and flow in capillaries and of microvessel diameter in the rat brain by confocal laser microscopy. AB - A new method for studying brain microcirculation is described. Both fluorescently labeled erythrocytes and plasma were visualized on-line through a closed cranial window in anesthetized rats, using laser-scanning two-dimension confocal microscopy. Video images of capillaries, arterioles, and venules were digitized off-line to measure microvessel diameter and labeled erythrocyte flow and velocity in parenchymal capillaries up to 200 microm beneath the brain surface. The method was used to analyze the rapid adaptation of microcirculation to a brief decrease in perfusion pressure. Twenty-second periods of forebrain ischemia were induced using the tour-vessel occlusion model in eight rats. EEG, arterial blood pressure, and body temperature were continuously controlled. In all conditions, labeled erythrocyte flow and velocity were both very heterogeneous in capillaries. During ischemia, capillary perfusion was close to 0, but a low blood flow persisted in arterioles and venules, while EEG was flattening. The arteriole and venule diameter did not significantly change. At the unclamping of carotid arteries, there was an instantaneous increase (by about 150%) of arteriole diameter. Capillary erythrocyte flow and velocity increased within 5 seconds, up to, respectively, 346 +/- 229% and 233 +/- 156% of their basal value. No capillary recruitment of erythrocytes was detected. All variables returned to their basal levels within less than 100 seconds after declamping. The data are discussed in terms of a possible involvement of shear stress in the reperfusion period. PMID- 10458594 TI - Simultaneous blood oxygenation level-dependent and cerebral blood flow functional magnetic resonance imaging during forepaw stimulation in the rat. AB - The blood oxygenation level-dependent (BOLD) contrast mechanism can be modeled as a complex interplay between CBF, cerebral blood volume (CBV), and CMRO2. Positive BOLD signal changes are presumably caused by CBF changes in excess of increases in CMRO2. Because this uncoupling between CBF and CMRO2 may not always be present, the magnitude of BOLD changes may not be a good index of CBF changes. In this study, the relation between BOLD and CBF was investigated further. Continuous arterial spin labeling was combined with a single-shot, multislice echo-planar imaging to enable simultaneous measurements of BOLD and CBF changes in a well-established model of functional brain activation, the electrical forepaw stimulation of alpha-chloralose-anesthetized rats. The paradigm consisted of two 18- to 30-second stimulation periods separated by a 1-minute resting interval. Stimulation parameters were optimized by laser Doppler flowmetry. For the same cross-correlation threshold, the BOLD and CBF active maps were centered within the size of one pixel (470 microm). However, the BOLD map was significantly larger than the CBF map. Measurements taken from 15 rats at 9.4 T using a 10-millisecond echo-time showed 3.7 +/- 1.7% BOLD and 125.67 +/- 81.7% CBF increases in the contralateral somatosensory cortex during the first stimulation, and 2.6 +/- 1.2% BOLD and 79.3 +/- 43.6% CBF increases during the second stimulation. The correlation coefficient between BOLD and CBF changes was 0.89. The overall temporal correlation coefficient between BOLD and CBF time courses was 0.97. These results show that under the experimental conditions of the current study, the BOLD signal changes follow the changes in CBF. PMID- 10458595 TI - The effect of transient global ischemia on the interaction of Src and Fyn with the N-methyl-D-aspartate receptor and postsynaptic densities: possible involvement of Src homology 2 domains. AB - Transient ischemia increases tyrosine phosphorylation of N-methyl-D-aspartate (NMDA) receptor subunits NR2A and NR2B in the rat hippocampus. The authors investigated the effects of this increase on the ability of the receptor subunits to bind to the Src homology 2 (SH2) domains of Src and Fyn expressed as glutathione-S-transferase-SH2 fusion proteins. The NR2A and NR2B bound to each of the SH2 domains and binding was increased approximately twofold after ischemia and reperfusion. Binding was prevented by prior incubation of hippocampal homogenates with a protein tyrosine phosphatase or by a competing peptide for the Src SH2 domain. Ischemia induced a marked increase in the tyrosine phosphorylation of several proteins in the postsynaptic density (PSD), including NR2A and NR2B, but had no effect on the amounts of individual NMDA receptor subunits in the PSD. The level of Src and Fyn in PSDs, but not in other subcellular fractions, was increased after ischemia. The ischemia-induced increase in the interaction of NR2A and NR2B with the SH2 domains of Src and Fyn suggests a possible mechanism for the recruitment of signaling proteins to the PSD and may contribute to altered signal transduction in the postischemic hippocampus. PMID- 10458596 TI - Increased sarcolemmal permeability in the cerebral artery during chronic spasm: an assessment using DNA-binding dyes and detection of apoptosis. AB - Alteration of sarcolemmal permeability was evaluated in the cerebral artery after subarachnoid hemorrhage. Significance of membrane dysfunction in the pathogenesis of chronic spasm and contribution of apoptosis were investigated in a canine model. Permeability of the smooth muscle cell (SMC) membrane was assessed by double staining with a hydrophilic (ethidium bromide [EB]) and a lipophilic (Hoechst 33342) DNA-binding dye. Quantitative observations were made with a ultraviolet-fluorescence microscope and a ultraviolet-laser confocal microscope. Occurrence of apoptosis was studied using electrophoresis and TUNEL method. In the normal arteries, nuclei of SMC were stained with Hoechst 33342 but not with EB. In the spastic arteries, SMC in the inner layer of the tunica media were stained with EB. The incidence of EB-positive cells reached maximum on day 7 (45 +/- 19%) and decreased in 2 to 4 weeks (13 +/- 5.2% and 5.0 +/- 2.1%, respectively), in parallel with amelioration of spasm. Electron and light microscopic observations revealed increased density of SMC cytoplasm with widening of the extracellular space. Necrosis was not evident. Apoptosis was not detected by the two methods. These results demonstrate that an augmentation in sarcolemmal permeability takes place during the course of chronic vasospasm and suggest its close correlation to pathogenesis. PMID- 10458597 TI - Induction of argininosuccinate synthetase in rat brain glial cells after striatal microinjection of immunostimulants. AB - The enzyme argininosuccinate synthetase (ASS) initiates the metabolic pathway leading from L-citrulline to L-arginine, the only physiological substrate of all isoforms of nitric oxide synthases. The presence of ASS in glial cells in vivo was investigated by immunohistochemical methods in a model of rat brain inflammation. Phosphate-buffered saline or a mixture of bacterial lipopolysaccharide and interferon-gamma was injected into the left striatum, and animals were killed 24 hours later. Ipsilateral and contralateral sides of brain sections were incubated with an antiserum against ASS or antibodies against cell specific markers. In the three areas examined, striatum, corpus callosum, and cortex, a strong induction of ASS immunoreactivity was observed in glial cells after injection of immunostimulants. A detailed quantitative analysis of double stained sections revealed that ASS was almost exclusively expressed in reactive, ED1-positive microglial cells/brain macrophages in immunostimulant- or sham injected ipsilateral sides of the sections. Furthermore, ASS/ED1 costaining was observed in perivascular cells. Colocalization of ASS with astroglial marker glial fibrillary acidic protein was given only occasionally after immunostimulation. ASS-positive neurons were detected in control and experimental animals; staining intensity was comparable in both cases. The results suggest that neurons express ASS constitutively, whereas the enzyme is induced in glial cells in response to proinflammatory stimuli. This finding is the first demonstration of an induction of a pathway auxiliary to generation of nitric oxide in brain in response to immunostimulants and provides new insight into neural arginine metabolism. PMID- 10458598 TI - Multiple microvascular and astroglial 5-hydroxytryptamine receptor subtypes in human brain: molecular and pharmacologic characterization. AB - Physiologic and anatomic evidence suggest that 5-hydroxytryptamine (5-HT) neurons regulate local cerebral blood flow and blood-brain barrier permeability. To evaluate the possibility that some of these effects occur directly on the blood vessels, molecular and/or pharmacologic approaches were used to assess the presence of 5-HT receptors in human brain microvascular fractions, endothelial and smooth muscle cell cultures, as well as in astroglial cells which intimately associate with intraparenchymal blood vessels. Isolated microvessels and capillaries consistently expressed messages for the h5-HT1B, h5-HT1D, 5-HT1F, 5 HT2A but not 5-HT7 receptors. When their distribution within the vessel wall was studied in more detail, it was found that capillary endothelial cells exhibited mRNA for the h5-HT1D and for the 5-HT7 receptors whereas microvascular smooth muscle cells, in addition to h5-HT1D and 5-HT7, also showed polymerase chain reaction products for h5-HT1B receptors. Expression of 5-HT1F and 5-HT2A receptor mRNAs was never detected in any of the microvascular cell cultures. In contrast, messages for all 5-HT receptors tested were detected in human brain astrocytes with a predominance of the 5-HT2A and 5-HT7 subtypes. In all cultures, sumatriptan inhibited (35-58%, P < .05) the forskolin-stimulated production of cyclic AMP, an effect blocked by the 5-HT1B/1D receptor antagonists GR127935 and GR55562. In contrast, 5-carboxamidotryptamine induced strong increases (> or = 400%, P < .005) in basal cyclic AMP levels that were abolished by mesulergine, a nonselective 5-HT7 receptor antagonist. Only astroglial cells showed a ketanserin sensitive increase (177%, P < .05) in IP3 formation when exposed to 5-HT. These results show that specific populations of functional 5-HT receptors are differentially distributed within the various cellular compartments of the human cortical microvascular bed, and that human brain astroglial cells are endowed with multiple 5-HT receptors. These findings emphasize the complex interactions between brain serotonergic pathways and non-neuronal cells within the CNS and, further, they raise the possibility that some of these receptors may be activated by antimigraine compounds such as brain penetrant triptan derivatives. PMID- 10458599 TI - Delayed hypovolemic hypotension exacerbates the hemodynamic and histopathologic consequences of thromboembolic stroke in rats. AB - Abnormalities in cerebrovascular reactivity or hemodynamic reserve are risk factors for stroke. The authors determined whether hemodynamic reserve is reduced in an experimental model of thromboembolic stroke. Nonocclusive common carotid artery thrombosis (CCAT) was produced in rats by a rose bengal-mediated photochemical insult, and moderate hypotension (60 mm Hg/30 min) was induced 1 hour later by hemorrhage. Alterations in local cerebral blood flow (ICBF) were assessed immediately after the hypotensive period by 14C-iodoantipyrine autoradiography, and histopathologic outcome was determined 3 days after CCAT. Compared to normotensive CCAT rats (n = 5), induced hypotension after CCAT (n = 7) led to enlarged regions of severe ischemia (i.e., mean ICBF < 0.24 mL/g/min) in the ipsilateral hemisphere. For example, induced hypotension increased the volume of severely ischemic sites from 16 +/- 4 mm3 (mean +/- SD) to 126 +/- 99 mm3 (P < 0.05). Histopathologic data also showed a larger volume of ischemic damage with secondary hypotension (n = 7) compared to normotension (22 +/- 15 mm3 versus 5 +/- 5 mm3, P < .05). Both hypotension-induced decreases in ICBF and ischemic pathology were commonly detected within cortical anterior and posterior borderzone areas and within the ipsilateral striatum and hippocampus. In contrast to CCAT, mechanical ligation of the common carotid artery plus hypotension (n = 8) did not produce significant histopathologic damage. Nonocclusive CCAT with secondary hypotension therefore predisposes the post-thrombotic brain to hemodynamic stress and structural damage. PMID- 10458601 TI - For better or worse? Telomerase inhibition and cancer. PMID- 10458600 TI - Effects of the AMPA receptor antagonist NBQX on outcome of newborn pigs after asphyxic cardiac arrest. AB - In neonates, asphyxia is a common cause of neuronal injury and often results in seizures. The authors evaluated whether blockade of alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) receptors during asphyxia and early recovery with 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo-(F)-quinoxaline (NBQX) ameliorates neurologic deficit and histopathology in 1-week-old piglets. Anesthetized piglets were exposed to a sequence of 30 minutes of hypoxia, 5 minutes of room air ventilation, 7 minutes of airway occlusion, and cardiopulmonary resuscitation. Vehicle or NBQX was administered intravenously before asphyxia (30 mg/kg) and during the first 4 hours of recovery (15 mg/kg/h). Neuropathologic findings were evaluated at 96 hours of recovery by light microscopic and cytochrome oxidase histochemical study. Cardiac arrest occurred at 5 to 6 minutes of airway occlusion, and cardiopulmonary resuscitation restored spontaneous circulation independent of treatment modalities in about 2 to 3 minutes. Neurologic deficit over the 96-hour recovery period was not ameliorated by NBQX. Seizure activity began after 24 to 48 hours in 7 of 10 animals with vehicle and in 9 of 10 of animals with NBQX. In each group, four animals died in status epilepticus. Neuropathologic outcomes were not improved by NBQX. The density of remaining viable neurons was decreased in parietal cortex and putamen by NBQX treatment. Metabolic defects in cytochrome oxidase activity were worsened by NBQX treatment. Seizure activity during recovery was associated with reduced neuronal viability in neocortex and striatum in piglets from both groups that survived for 96 hours. This neonatal model of asphyxic cardiac arrest and resuscitation generates neurologic deficits, clinical seizure activity, and selective damage in regions of basal ganglia and sensorimotor cortex. In contrast to other studies in mature brain, AMPA receptor blockade with NBQX failed to protect against neurologic damage in the immature piglet and worsened postasphyxic histopathologic outcome in neocortex and putamen. PMID- 10458602 TI - Sex, cell death, and the genome of C. elegans. PMID- 10458603 TI - Perspectives on oxygen sensing. PMID- 10458604 TI - Twenty-five years of the nucleosome, fundamental particle of the eukaryote chromosome. PMID- 10458605 TI - Stat3 as an oncogene. AB - STATs are latent transcription factors that mediate cytokine- and growth factor directed transcription. In many human cancers and transformed cell lines, Stat3 is persistently activated, and in cell culture, active Stat3 is either required for transformation, enhances transformation, or blocks apoptosis. We report that substitution of two cysteine residues within the C-terminal loop of the SH2 domain of Stat3 produces a molecule that dimerizes spontaneously, binds to DNA, and activates transcription. The Stat3-C molecule in immortalized fibroblasts causes cellular transformation scored by colony formation in soft agar and tumor formation in nude mice. Thus, the activated Stat3 molecule by itself can mediate cellular transformation and the experiments focus attention on the importance of constitutive Stat3 activation in human tumors. PMID- 10458606 TI - Nuclear trafficking of Cubitus interruptus in the transcriptional regulation of Hedgehog target gene expression. AB - Transcriptional activation of Hedgehog (Hh) target genes requires Cubitus interruptus, a 155 kDa cytoplasmic zinc finger protein (Ci155), which in the absence of Hh signaling is processed to form a nuclear repressor (Ci75). We show that Hh signaling reduces phosphorylation of Ci155, and this reduction in turn appears to decrease processing. Blocking processing with proteasome inhibitors or altered Ci proteins, however, is insufficient for activation of Hh targets. We find that Hh signaling increases the rate of Ci155 nuclear import, resulting in significant nuclear accumulation. Even in the absence of signaling, nuclear accumulation of Ci155 suffices for significant induction of Hh targets, and active nuclear export of Ci155 is an essential mechanism for maintenance of the unstimulated state. PMID- 10458607 TI - The TRA-1A sex determination protein of C. elegans regulates sexually dimorphic cell deaths by repressing the egl-1 cell death activator gene. AB - The hermaphrodite-specific neurons (HSNs) of the nematode Caenorhabditis elegans are generated embryonically in both hermaphrodites and males but undergo programmed cell death in males. The gene egl-1 encodes a BH3-containing cell death activator that is required for programmed cell death in C. elegans. Gain-of function (gf) mutations in egl-1 cause the inappropriate programmed cell death of the HSNs in hermaphrodites. These mutations lie 5.6 kb downstream of the egl-1 transcription unit and disrupt the binding of the TRA-1A zinc finger protein, the terminal global regulator of somatic sexual fate. This disruption results in the activation of the egl-1 gene in the HSNs not only in males but also in hermaphrodites. Our findings suggest that in hermaphrodites TRA-1A represses egl 1 transcription in the HSNs to prevent these neurons from undergoing programmed cell death. PMID- 10458608 TI - Identification and cloning of a negative regulator of systemic acquired resistance, SNI1, through a screen for suppressors of npr1-1. AB - Systemic acquired resistance (SAR) is a plant immune response induced after a local infection by necrotizing pathogens. The Arabidopsis NPR1 gene is a positive regulator of SAR, essential for transducing the SAR signal salicylic acid (SA). Mutations in the NPR1 gene abolish the SA-induced expression of pathogenesis related (PR) genes and resistance to pathogens. To identify additional regulators of SAR, we screened for suppressors of npr1-1. In the npr1-1 background, the sni1 (suppressor of npr1-1, inducible 1) mutant shows near wild-type levels of PR1 expression and resistance to pathogens after induction. Restoration of SAR in npr1-1 by the recessive sni1 mutation indicates that wild-type SNI1 may function as a negative regulator of SAR. We cloned the SNI1 gene and found that it encodes a leucine-rich nuclear protein. PMID- 10458609 TI - Giant eyes in Xenopus laevis by overexpression of XOptx2. AB - Overexpression of XOptx2, a homeodomain-containing transcription factor expressed in the Xenopus embryonic eye field, results in a dramatic increase in eye size. An XOptx2-Engrailed repressor gives a similar phenotype, while an XOptx2-VP16 activator reduces eye size. XOptx2 stimulates bromodeoxyuridine incorporation, and XOptx2-induced eye enlargement is dependent on cellular proliferation. Moreover, retinoblasts transfected with XOptx2 produce clones of cells approximately twice as large as control clones. Pax6, which does not increase eye size alone, acts synergistically with XOptx2. Our results suggest that XOptx2, in combination with other genes expressed in the eye field, is crucially involved in the proliferative state of retinoblasts and thereby the size of the eye. PMID- 10458610 TI - Regulation of DLG localization at synapses by CaMKII-dependent phosphorylation. AB - Discs large (DLG) mediates the clustering of synaptic molecules. Here we demonstrate that synaptic localization of DLG itself is regulated by CaMKII. We show that DLG and CaMKII colocalize at synapses and exist in the same protein complex. Constitutively activated CaMKII phenocopied structural abnormalities of dlg mutant synapses and dramatically increased extrajunctional DLG. Decreased CaMKII activity caused opposite alterations. In vitro, CaMKII phosphorylated a DLG fragment with a stoichiometry close to one. Moreover, expression of site directed dlg mutants that blocked or mimicked phosphorylation had effects similar to those observed upon inhibiting or constitutively activating CaMKII. We propose that CaMKII-dependent DLG phosphorylation regulates the association of DLG with the synaptic complex during development and plasticity, thus providing a link between synaptic activity and structure. PMID- 10458611 TI - The sleep disorder canine narcolepsy is caused by a mutation in the hypocretin (orexin) receptor 2 gene. AB - Narcolepsy is a disabling sleep disorder affecting humans and animals. It is characterized by daytime sleepiness, cataplexy, and striking transitions from wakefulness into rapid eye movement (REM) sleep. In this study, we used positional cloning to identify an autosomal recessive mutation responsible for this sleep disorder in a well-established canine model. We have determined that canine narcolepsy is caused by disruption of the hypocretin (orexin) receptor 2 gene (Hcrtr2). This result identifies hypocretins as major sleep-modulating neurotransmitters and opens novel potential therapeutic approaches for narcoleptic patients. PMID- 10458612 TI - Oligomeric complexes link Rab5 effectors with NSF and drive membrane fusion via interactions between EEA1 and syntaxin 13. AB - SNAREs and Rab GTPases cooperate in vesicle transport through a mechanism yet poorly understood. We now demonstrate that the Rab5 effectors EEA1 and Rabaptin 5/Rabex-5 exist on the membrane in high molecular weight oligomers, which also contain NSF. Oligomeric assembly is modulated by the ATPase activity of NSF. Syntaxin 13, the t-SNARE required for endosome fusion, is transiently incorporated into the large oligomers via direct interactions with EEA1. This interaction is required to drive fusion, since both dominant-negative EEA1 and synthetic peptides encoding the FYVE Zn2+ finger hinder the interaction and block fusion. We propose a novel mechanism whereby oligomeric EEA1 and NSF mediate the local activation of syntaxin 13 upon membrane tethering and, by analogy with viral fusion proteins, coordinate the assembly of a fusion pore. PMID- 10458613 TI - The protein CTCF is required for the enhancer blocking activity of vertebrate insulators. AB - An insulator is a DNA sequence that can act as a barrier to the influences of neighboring cis-acting elements, preventing gene activation, for example, when located between an enhancer and a promoter. We have identified a 42 bp fragment of the chicken beta-globin insulator that is both necessary and sufficient for enhancer blocking activity in human cells. We show that this sequence is the binding site for CTCF, a previously identified eleven-zinc finger DNA-binding protein that is highly conserved in vertebrates. CTCF sites are present in all of the vertebrate enhancer-blocking elements we have examined. We suggest that directional enhancer blocking by CTCF is a conserved component of gene regulation in vertebrates. PMID- 10458614 TI - Structure of the DNA repair enzyme endonuclease IV and its DNA complex: double nucleotide flipping at abasic sites and three-metal-ion catalysis. AB - Endonuclease IV is the archetype for a conserved apurinic/apyrimidinic (AP) endonuclease family that primes DNA repair synthesis by cleaving the DNA backbone 5' of AP sites. The crystal structures of Endonuclease IV and its AP-DNA complex at 1.02 and 1.55 A resolution reveal how an alpha8beta8 TIM barrel fold can bind dsDNA. Enzyme loops intercalate side chains at the abasic site, compress the DNA backbone, bend the DNA approximately 90 degrees, and promote double-nucleotide flipping to sequester the extrahelical AP site in an enzyme pocket that excludes undamaged nucleotides. These structures suggest three Zn2+ ions directly participate in phosphodiester bond cleavage and prompt hypotheses that double nucleotide flipping and sharp bending by AP endonucleases provide exquisite damage specificity while aiding subsequent base excision repair pathway progression. PMID- 10458615 TI - Failure of HIV-1 to infect human oocytes directly. AB - In this study, the susceptibility of mature human oocytes to HIV-1 infection has been investigated. We exposed in vitro human oocytes of healthy women using inocula of cell-free HIV-1. We also tested for the presence of HIV-1-specific receptor molecules on the surface of these cells. By applying polymerase chain reaction (PCR) analysis, transmission electron microscopy (TEM), and immunocytochemistry at both light and electron microscopic levels, we did not obtain evidence for HIV DNA production nor for oocyte-associated HIV particles. Experiments of immunostaining for CD4, CCR5, and GalAAG (putative receptor for HIV in sperm), as well as reverse transcriptase (RT)-PCR for CD4, CCR5, and CXCR4, which all suggested the absence of the mentioned receptors in mature oocytes and in follicular cells. This study fills an important gap concerning the information available on the direct HIV infection of human gametes, adds to our basic understanding of HIV infection in human oocytes, provides different results from those obtained with human spermatozoa using comparable methods, and provides a basic contribution to the investigation on HIV infection in human oocytes. PMID- 10458616 TI - Abacavir and mycophenolic acid, an inhibitor of inosine monophosphate dehydrogenase, have profound and synergistic anti-HIV activity. AB - The use of inhibitors of purine nucleoside metabolism has been advocated for the treatment of HIV-1 infection. Abacavir is the first clinically available guanosine analogue HIV-1 reverse transcriptase inhibitor, and the most potent nucleoside analogue yet developed. Mycophenolic acid (MA), a specific inhibitor of lymphocyte proliferation that is currently in use in organ transplantation, acts on inosine monophosphate dehydrogenase to block conversion of inosine monophosphate to guanosine monophosphate. We found abacavir and MA inhibited HIV 1 replication in stimulated peripheral blood mononuclear cells (PBMCs) and in monocyte-derived macrophages (MDMs). Inhibition was potent and synergistic to an extent not previously observed with other antiretroviral combinations. MA was effective at concentrations (0.25 microM) far below those used for immunosuppression in organ transplantation. An HIV strain encoding the M184V mutation was susceptible to the combination of MA and abacavir. However, the combination of MA and zidovudine (ZDV) or stavudine (d4T) was antagonistic. Although the translation of these observations must be carefully evaluated in clinical trials, the judicious combination of antiretrovirals and inhibitors of nucleoside metabolism may emerge as an important strategy in the treatment of HIV infection. PMID- 10458617 TI - Cerebrospinal fluid HIV RNA and drug levels with combination ritonavir and saquinavir. AB - Combination antiretroviral therapy with ritonavir and saquinavir has established potent and durable activity on plasma viremia. CNS HIV infection may be sequestered from drug therapy that does not penetrate the blood-brain barrier. Penetration of these protease inhibitors into the cerebrospinal fluid (CSF) and CSF HIV RNA levels on such therapy has not been well described. DESIGN/METHODS: In a cross-sectional study, 28 HIV1-infected study subjects were evaluated either before initiation of or before maximal response to ritonavir-saquinavir therapy, during maximal plasma virologic response, and after virologic failure. Simultaneous samples of plasma and cerebrospinal fluid were obtained from 24 study subjects to measure HIV RNA and protease inhibitor levels. RESULTS: Across the treatment groups, a strong correlation was found between plasma and CSF HIV RNA levels (r = 0.870; p < .001). In each study subject with plasma HIV RNA levels below assay limit (80 copies/ml), the CSF HIV RNA level was also below the limit of quantitation. Low levels of saquinavir (<2 ng/ml) and ritonavir (<25 ng/ml) in the CSF were observed, with a CSF:plasma drug concentration ratio of < or = 0.005 (0.5%) in all study subjects evaluated (n = 11). The plasma:CSF HIV RNA ratio was high before or early in treatment (median, 38; interquartile range [IQR], 13,97), but low (median, 0.29; IQR, 0.17, 7.5) in those failing therapy (group C, p < .001). CONCLUSIONS: CSF ritonavir and saquinavir levels are consistent with the estimated known fraction of unbound drug in plasma (<2%). Across these treatment response groups, suppression of plasma viremia can predict low CSF HIV RNA levels. This correlation may represent HIV RNA transport and equilibrium between CSF and plasma, or it may represent CNS anti-HIV activity of protease inhibitors. The low drug levels and inverted ratio of HIV RNA in the CSF compared with plasma early in plasma virologic breakthrough suggests CSF virologic failure may contribute to failure of plasma virologic response. PMID- 10458618 TI - Immune function and phenotype before and after highly active antiretroviral therapy. AB - Immune functions represented by equal CD4 counts before and after highly active antiretroviral therapy (i.e., pre- and post-HAART) in the same HIV-infected patients, were examined. Twelve HIV-infected patients were included. Patients had equal CD4 counts pre- and post-HAART and were studied on average 30 months pre HAART and 17 months post-HAART. Post-HAART, CD8+ T cells expressed greater amounts of CD28 (p < .02), smaller amounts of CD38 (p < .02), and a reduced proportion of CD4+CD28+ T cells expressed CD38+ (p < .01). Proliferation increased (p < 10) in lymphocyte cell cultures stimulated with pokeweed mitogens or Candida, and was correlated to expression of CD28 on T cells (p < .02). The proportion of CD3-CD16-CD56+ natural killer (NK) cells increased (p < .05) and CD3-CD16+CD56- NK cells declined (p < .01). Production of interferon-gamma increased (p < .10). The number of naive and memory T cells, the non-major histocompatibility complex (non-MHC)-restricted and HIV-specific MHC-restricted cytotoxicity and the production of macrophage inflammatory protein-1gamma were unchanged. The finding of increased expression of CD28, correlating to increased proliferation capacity, and diminished expression of CD38 on T cells, indicates that following long-term HAART, repopulation occurs with less activated cells with increased proliferative capacity. This finding may be of clinical importance in considering risk and vulnerability for progression of opportunistic infections post-HAART. PMID- 10458619 TI - Antiviral therapy for HIV patients with renal insufficiency. AB - Patients with HIV infection and HIV-related opportunistic infections are treated extensively with a spectrum of drugs. Introduction of new antiretroviral drugs, such as protease inhibitors and nonnucleoside reverse transcriptase inhibitors in addition to nucleoside reverse transcriptase inhibitors, has created exciting dimensions in treatment strategies. Renal dysfunction is also common in HIV infected patients. Because some drugs used in HIV are primarily excreted unchanged by the kidney, dose adjustments are necessary in patients with renal insufficiency. Drugs such as foscarnet, cidofovir and adefovir are directly nephrotoxic, whereas acyclovir can crystallize in the kidneys, and indinavir may cause nephrolithiasis. This paper reviews the impact of renal insufficiency on pharmacokinetics of antiviral drugs used in HIV disease and discusses dosage recommendations needed to avoid toxicity. Finally, we summarize the effects of dialysis on removal of these drugs. PMID- 10458620 TI - Obstacles to needle exchange participation in Rhode Island. AB - OBJECTIVE: This study explores obstacles to participation in needle exchange programs (NEPs) among injection drug users (IDUs) in the state of Rhode Island, U.S.A. METHODS: A written questionnaire was administered at two Rhode Island drug detoxification sites in 1998. RESULTS: 488 self-administered surveys were completed, 226 (46.3%) respondents had injected drugs in the past 6 months. 62.1% reported sharing syringes in the past 6 months, and each syringe was used a mean of 10.7 times. Major obstacles to NEP participation were a lack of awareness of the program (25.6%), inconvenient location or hours (15.9%), and fear of identification and/or police harassment (12.2%). Non-white race was a significant predictor of being unaware of the NEP (p = .01) and not participating in the NEP (p = .03). 13.1% of IDUs who used the NEP were referred to the detoxification program by the NEP. Among all IDUs surveyed, 51.0% had participated in a NEP. CONCLUSIONS: NEPs are important in reducing the spread of bloodborne pathogens among IDUs and are effective referral sources for drug treatment. Surveys of IDUs at sites other than NEPs, such as detoxification facilities, can identify obstacles to the use of NEPs. PMID- 10458621 TI - Changes in AIDS-defining illnesses in a London Clinic, 1987-1998. AB - OBJECTIVES: To describe the incidence of AIDS-defining illnesses within a single large clinic setting, to describe temporal changes over a 10-year period in the overall incidence and of individual AIDS-defining illnesses and to investigate the impact of HIV treatment regimen on the incidence of AIDS-defining illnesses. SUBJECTS AND METHODS: A person-years analysis was used to determine the incidence of AIDS-defining illnesses according to calendar year and stratification by CD4 lymphocyte count and treatment regimen in 1806 patients from the Royal Free Centre for HIV Medicine with at least one CD4 lymphocyte count and follow-up visit. RESULTS: Prior to 1992, the incidence of all AIDS-defining illnesses was 27.4/100 person-years of follow-up (PYFU; 95% confidence interval [CI], 22.8 32.0) and during 1997 this incidence had dropped to 6.9/100 PYFU (95% CI, 4.7 9.1; p < .0001, test for trend). The decline in incidence over time occurred in esophageal candidiasis, cytomegalovirus disease (including retinitis), Kaposi's sarcoma, lymphoma, wasting syndrome, and Pneumocystis carinii pneumonia (p < .05, test for trend), but there was no evidence of a decline in AIDS dementia or in Mycobacterium avium complex. In 1997, among patients with CD4 lymphocyte counts of < or =200 cells/mm3, the incidence rates for any AIDS-defining illness was 51.1/100 PYFU for patients taking no therapy (95% CI, 27.9-85.7), 34.5 among patients on monotherapy (95% CI, 4.2-124.6), 13.2 among patients taking dual combination therapy (95% CI, 3.6-33.8) and 6.1 among patients taking triple therapy or more complex regimens (95% CI, 0.7-22.0; p < .0001, test for trend). CONCLUSIONS: There was a considerable decline in AIDS-defining illnesses during 1996 and 1997, coinciding with the rapid development of new antiretroviral treatments and combinations of treatment. Further follow-up of large observational cohorts is essential to monitor the incidence of diagnoses less common than we were able to consider, such as tuberculosis, cryptosporidiosis, and cryptococcosis, and also to investigate whether the incidence of disease continues to fall, or whether it starts to rise again, as toxicities, compliance, drug resistance, and long-term side effects begin to appear. PMID- 10458623 TI - Belief of vaccine receipt in HIV vaccine trials: further cautions. AB - The purpose of this cross-sectional study was to examine relationships between belief in vaccine receipt, motivations for trial participation, and side effects in phase 1 vaccine trials. Anonymous questionnaires were completed by 125 active vaccine volunteers at two vaccine evaluation sites. Participants believing they had received the vaccine reported more side effects (p < .01), were less likely to report knowing someone with HIV/AIDS as a motivation for trial participation (p < .01), and endorsed greater concern about becoming HIV-infected as motivation for participation (p < .05). Results indicate that inferences made by trial participants in vaccine trials should be identified and addressed, and that greater efforts for maintaining the blinded nature of vaccine trials and educating trial participants about the meaning of side effects are warranted. PMID- 10458622 TI - Incident HIV infection in a high-risk, homosexual, male cohort in Rio de Janeiro, Brazil. AB - An HIV seroincidence study was conducted to identify a high-risk population for HIV prevention trials. Inclusion criteria were male gender, homosexual behavior, age between 18 and 50 years, and negative HIV serostatus; 862 study subjects were screened and 753 were enrolled and observed during follow-up for a mean of 1.5 years. In this population, 34 people had HIV seroconversions for an overall annual seroincidence of 3.1% (95% confidence interval [CI], 2.1%-4.1%). Among study subjects <20 years old, annual incidence was 8.4% (95% CI, 1.7%-15%). Independent risk factors for seroconversion were age <25 years (p = .01), hepatitis B core antibody seropositivity (p > .01), sex at first encounter in the preceding 6 months (p = .11), and a history of gonorrhea or condyloma in the 6 months before seroconversion (p = .04 and p = .08, respectively). At enrollment, 85% of the eventual seroconverters said they would participate in a vaccine trial; all agreed to participate when told there would be a placebo arm. Follow up rates were 97%, 91%, and 88% at 6, 12, and 18 months, respectively. The HIV-1 subtype was B for each of the first 17 seroconverters. These data demonstrate the suitability of this cohort for HIV prevention trials, based on high HIV incidence and retention rates, and a willingness to participate in such trials. PMID- 10458624 TI - Home collection versus publicly funded HIV testing in San Francisco: who tests where? AB - We examined records of all HIV antibody tests performed at anonymous publicly funded (PF) sites and by home collection (HC) testing for residents of San Francisco from August 1996 to December 1997. Although far fewer tests were performed by HC testing than at PF sites (715 versus 8712, respectively), a higher proportion of HC testers reported no prior history of HIV testing (33.1% versus 17.9%). HIV seroprevalence was higher among PF tests (1.8%) than among HC tests (0.9%). Compared with PF testers, HC testers were less likely to be gay men, lesbian or bisexual women, heterosexual women, African American, or Latino. HC testers were more likely to report sex with a known HIV-positive partner. HC testers were also more likely to reside in affluent neighborhoods. HC testing reaches some high-risk persons who may not otherwise seek PF testing, although, overall, the risk profile of HC testers appeared lower than that of PF testers. HC testing reaches some individuals who can financially afford HC testing, thus saving public prevention resources for hard-to-reach, high-risk populations. PMID- 10458625 TI - Lipid abnormalities during saquinavir soft-gel-based highly active antiretroviral therapy. PMID- 10458626 TI - Hydroxyurea as part of a salvage regimen for heavily pretreated patients with advanced HIV infection. PMID- 10458627 TI - Inappropriate model-fitting methods may lead to significant underestimates of viral decay rates in HIV dynamic studies. PMID- 10458628 TI - A phase-II study of trovafloxacin for the treatment of Chlamydia trachomatis infections. AB - BACKGROUND: Chlamydia trachomatis can cause an insidious, asymptomatic infection in both men and women. Trovafloxacin, a fluoronaphthyridone antibiotic related to the fluoroquinolones, has broad antibacterial activity that encompasses C. trachomatis. GOAL OF STUDY: We designed a phase-II study to determine the efficacy and safety of trovafloxacin for chlamydial infections. STUDY DESIGN: Four once-daily dosing regimens were used: 200 mg for 7 days, 200 mg for 5 days, 100 mg for 7 days, and 50 mg for 7 days. RESULTS: Sixty-four men and 66 women were enrolled. Of the 73 bacteriologically evaluable patients, 20 received 200 mg for 7 days, 18 received 200 mg for 5 days, 14 received 100 mg for 7 days, and 21 received 50 mg for 7 days. Bacteriologic failure occurred in 1 patient (5%) in the 200-mg 7-day group, 2 patients (14%) in the 100-mg group, and 1 patient (5%) in the 50-mg group. No clinical failures were noted among the 30 C. trachomatis infected men with nongonococcal urethritis, although 1 patient had only improved at the final visit. Safety data were available from 119 patients. Of 86 patients receiving the three highest dosing regimens, 17 (20%) had at least one adverse event, judged treatment-related in only 9 (10 %). Of 33 patients receiving the lowest dose, 5 (15%) had an adverse event, three (9%) of which were considered treatment related. No adverse event was severe. CONCLUSION: In doses ranging from 200 mg to 50 mg daily for a week and 200 mg daily for 5 days, trovafloxacin appears to be effective for chlamydial infections and is well tolerated. PMID- 10458629 TI - Recording sexual behavior: comparison of recall questionnaires with a coital diary. AB - OBJECTIVE: To compare sexual behavior data obtained using a weekly-recall questionnaire, a daily-recall questionnaire, and a coital diary. DESIGN: Cross sectional survey of female sex workers from KwaZulu-Natal, South Africa between August and October 1998. METHODS: In this study, 52 weekly-recall questionnaires, 27 daily-recall questionnaires, and 79 coital diaries for dates corresponding to the questionnaires were obtained from female sex workers. Variables examined included: number of clients, number of condoms used with clients and partners, and number and type of sexual acts with clients and partners. Statistical analyses were conducted to examine the degree of agreement between the data collection methods and to assess differences between the mean values of the variables in the questionnaires versus the diary. RESULTS: Comparison of weekly recall questionnaires with coital diaries indicated a significantly greater mean number of clients (P < 0.001), number of condoms used (P < 0.001), vaginal acts (P < 0.001), and anal acts (P = 0.044) reported in the diary versus the questionnaire. On comparison of daily-recall questionnaire with coital diary, significant differences were revealed between the means detected for the number of clients (P = 0.027), number of days worked (0.009), and anal acts with clients (P = 0.004). CONCLUSIONS: The use of coital diaries for the collection of sexual behavior data may be limited to cross-sectional community surveys. A recall questionnaire may provide more reliable and a better quality of data for longitudinal studies and for human immunodeficiency virus/sexually transmitted disease evaluation programs. PMID- 10458630 TI - Trichomonas vaginalis as a cause of urethritis in Malawian men. AB - BACKGROUND AND OBJECTIVES: Trichomonas vaginalis is one of the most common sexually transmitted infections. In Malawi, rates of trichomoniasis in women are high. The prevalence of T. vaginalis infection in men is expected to be high but has not previously been documented. GOALS: We sought to determine the prevalence of trichomoniasis in Malawian men with and without urethritis, to evaluate a polymerase chain reaction detection assay for T. vaginalis in urethral swabs and to examine the effect of T. vaginalis infection on excretion of human immunodeficiency virus (HIV) in semen. STUDY DESIGN: Men presenting at the Sexually Transmitted Diseases (STD) and Dermatology Clinics in Malawi were enrolled in a cross-sectional study. We compared a polymerase chain reaction based test for T. vaginalis detection with wet-mount microscopy and culture of urethral swabs. HIV serology was determined by enzyme-linked immunosorbent assay (ELISA), and HIV-1 RNA concentrations in semen were measured by quantitative nucleic acid sequence-based analysis. RESULTS: T. vaginalis was detected in 51 of 293 men. The estimated prevalence among symptomatic men was 20.8% and among asymptomatic men, 12.2%. Polymerase chain reaction performed with a sensitivity of 0.82 (95% CI: 0.66-0.92) and specificity of 0.95 (95% CI: 0.91-0.97) compared to wet-mount microscopy and culture. There was no difference in the rate of HIV seropositivity in men with and without T. vaginalis infection. However, in men with symptomatic urethritis, the median HIV RNA concentration in seminal plasma from men with T. vaginalis was significantly higher that in seminal plasma from HIV-positive men without trichomonas. PMID- 10458631 TI - Trichomonas vaginalis--no longer a minor STD. PMID- 10458632 TI - Reemergence of gonorrhea in Sweden. PMID- 10458633 TI - Clinical features of Chlamydia trachomatis rectal infection by serovar among homosexually active men. AB - BACKGROUND AND OBJECTIVES: Because C. trachomatis serovars correlate with the clinical manifestations of cervical infection, we undertook this study to determine whether clinical, behavioral, and laboratory findings correlate with C. trachomatis serovars isolated from rectal infections. GOAL OF THIS STUDY: To correlate C. trachomatis serovar with signs and symptoms of rectal infection. STUDY DESIGN: A cross-sectional study of 454 men with rectal C. trachomatis infection attending an urban sexually transmitted disease (STD) clinic was undertaken. Isolates were thawed, passaged to high titer, and typed using a panel of monoclonal antibodies. Compared to men infected with B complex isolates (164), men with C complex isolates (55) were less likely to report symptoms (OR: 0.4; 95% CI: 0.1-0.8), or to have erythema, bleeding, or mucopus (OR: 0.3; 95% CI: 0.1 0.8). Among men with inclusion counts of more than 100, those infected with FG group versus B complex isolates were more likely to present with mucopus (OR: 10.5; 95% CI: 1.2-95.5), more than 15 polymorphonuclear leukocytes (OR: 19.2; 95% CI: 1.7-219.8), and proctitis (OR: 4.2; 95% CI: 1.1-16.7). CONCLUSION: Signs and symptoms of rectal infection correlate with the serovar of C. trachomatis isolates. PMID- 10458634 TI - Perceived monogamy and type of clinic as barriers to seeking care for suspected STD or HIV infection: results from a brief survey of low-income women attending Women, Infants, and Children (WIC) clinics in Missouri. AB - BACKGROUND AND OBJECTIVES: Barriers to seeking care for sexually transmitted diseases (STDs) have not been assessed for low-income women. We sought to determine barriers to seeking care for STDs among women receiving Women, Infants, and Children (WIC) benefits in 21 Missouri counties. GOAL OF THIS STUDY: Provide information for promoting care-seeking behavior among low-income women suspecting STD infection. STUDY DESIGN: A survey of 2,256 women was conducted; 491 reported a history of at least one STD. These women indicated possible barriers to seeing a doctor about a suspected STD and preference for type of clinic providing STD services. RESULTS: More than one fifth (21.3%) of those reporting an STD also reported at least one barrier to seeing a doctor about suspected STD or human immunodeficiency virus infection. Among those reporting barriers, the most common barrier was "I only have sex with my steady" (36.2%) followed by being asymptomatic (33.3%), embarrassment (22.8%), and cost (25.7%). Most (63.8%) preferred seeing their own doctor, with others reporting preference for community health centers (14.8%), family planning clinics (16.8%), and STD clinics (4.6%). CONCLUSIONS: Low-income women experience multiple barriers to seeking care including perceptions about a protective value of monogamy. Also, STD services in locations providing other health services for women were preferred. PMID- 10458635 TI - Bacterial vaginosis, ethnicity, and the use of genital cleaning agents: a case control study. AB - BACKGROUND AND OBJECTIVES: Bacterial vaginosis and vaginal douching are both reported to be more common in African-American and Caribbean than white women. It is also thought that douching alters the vaginal milieu. This study was conducted to examine associations between genital cleaning practices, bacterial vaginosis, and ethnic group. STUDY DESIGN: Case-control study of 100 women with bacterial vaginosis, diagnosed by Nugent's criteria, and 100 women without bacterial vaginosis attending a sexually transmitted diseases clinic in an ethnically heterogeneous inner-city area in London, England. RESULTS: Bacterial vaginosis was more common among black Caribbean than white women (OR, 2.1; 95% CI, 1.1 4.1). Vulval use of bubble bath or antiseptic solutions and douching with proprietary or homemade solutions were significantly more common in women with bacterial vaginosis than without. After controlling for use of vulval and vaginal antiseptics and bubble bath, douching, and a history of bacterial vaginosis, there was no ethnic difference in the occurrence of the condition (adjusted OR, 1.1; 95% CI, 0.5-2.5). CONCLUSIONS: Ethnic differences in genital hygiene behaviors can explain a twofold increase in the risk of bacterial vaginosis in black Caribbean compared with white women. The role of vulval and vaginal cleaning practices in the development of bacterial vaginosis should be examined further in longitudinal or randomized controlled studies. PMID- 10458636 TI - Prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and HIV infection among drug users attending an STD/HIV prevention and needle-exchange program in Quebec City, Canada. AB - OBJECTIVES: To determine the prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and human immunodeficiency virus (HIV) infection among injection and noninjection drug users in Quebec City and to identify associated risk factors. STUDY DESIGN: Cross-sectional study of 738 drug users attending a sexually transmitted disease/HIV prevention and needle-exchange program in Quebec City, Canada. RESULTS: The prevalences of N. gonorrhoeae and C. trachomatis were, respectively, 0.4% (95% CI: 0.08%-1.2%) and 3.4% (95% Cl: 2.2%-5.0%). Through multivariate analysis, risk factors associated with sexually transmitted diseases were: among women, being aged between 20 and 24 years and having unprotected sex with commercial partners; in noninjection drug users; cocaine use and first intercourse before 13 years of age; in male noninjection drug users, having regular sexual partners in the previous 6 months. No case of HIV was found in participants who have never injected drugs, but the prevalence was 5.5% (6/110) among the exinjectors and 10.1% (35/347) in current injectors. CONCLUSION: Moderate sexually transmitted disease and HIV prevalences were found, although a high proportion of drug users reported risky behaviors. Needle-exchange program sites may offer a good opportunity to provide sexually transmitted disease/HIV prevention and medical services to drug users. PMID- 10458637 TI - Laser light scattering immunoassay for malaria. AB - Laser light scattering immunoassay (LIA) was proposed as a prospective diagnostic method for the detection of antibody (or antigen) by monitoring the agglutination of antigen (or antibody) coated carrier particles using dynamic light scattering (DLS) as probe. LIA is a very sensitive assay as it can detect microscopic immune complexes even when antibody (or antigen) level is low. A sizeable number of human sera collected from malaria endemic areas and hospitals have been analysed by ELISA using Pf parasite lysate or a RESA derived synthetic peptide as antigen parallel to LIA using Pf antigen coated polystyrene latex beads. Comparative analysis of data suggests LIA to be as good as ELISA and possibly better in terms of sensitivity and simplicity. LIA can be a simple and inexpensive immunoassay suitable for field use and mass application. PMID- 10458638 TI - Fluorescence polarization assay: application to the diagnosis of bovine brucellosis in Argentina. AB - A homogeneous fluorescence polarization assay (FPIA) for detection of bovine antibody to Brucella abortus was validated in Argentina Sera were defined based on their reactivity in the buffered antigen plate agglutination test (BPAT) and the competitive enzyme immunoassay (CELISA). Sera negative in these tests were collected from farms without evidence of brucellosis (n=733). Sera positive in the two tests were collected from cattle on farms from which B. abortus was isolated from at least one animal (n=1039). Sera from cattle vaccinated 26, 89, 240 and 272 days previously with B. abortus strain 19 were collected and tested. A cut-off value of 87 mP was determined for the FPIA, resulting in relative sensitivity and specificity values of 98.1 and 99.6%. The specificity for B. abortus strain 19 vaccinated cattle was 64.9% (26 days post vaccination, DPV), 92.1% (89 DPV), 98.6% (242 DPV) and 97.1% (272 DPV). These values were compared to those obtained with the BPAT, the CELISA, the indirect ELISA, the complement fixation test and the 2-mercaptoethanol agglutination test. Sera from 18 cattle which were vaccinated and revaccinated with B. abortus strain 19 were also tested by the same assays and the FPIA was found to be 100% specific. The use of the FPIA as a diagnostic test for brucellosis is discussed. PMID- 10458639 TI - Detection of zooplankton prey in squid paralarvae with immunoassay. AB - Sustainable management of economically important squid requires monitoring of changes in their abundance, which are related inter alia, to their success in the food chain. The highest mortality is expected in the paralarval stages, which are prone to starvation. Causes of starvation may be linked to the lack of suitable prey. A multiple detection system was developed for the simultaneous identification of five putative zooplankton prey in the guts of paralarval Chokka squid, Loligo vulgaris reynaudii, by employing polyclonal rabbit antisera in conjunction with solid phase immunoassays. Specificities of antisera were validated by ELISA screening against different zooplankton taxa. Cross-reactions observed with ELISA were minimized through manipulation of antibody and antigen concentrations resulting in more specific detection of target prey antigens when used in an immunodot assay. Application of this optimised immunoassay detected multiple predation in paralarval squid samples collected from diverse areas in the Agulhas Bank ecosystem on the south coast of South Africa. PMID- 10458640 TI - A comparison of performances of four enzymes used in ELISA with special reference to beta-lactamase. AB - Horse radish peroxidase, alkaline phaosphatase and beta-D-galactosidase are widely used as labels in the development of enzyme immunoassays (EIAs). Enzyme beta-lactamase, though introduced as a label in late seventies has not yet become very popular inspite of having the necessary features of an enzyme to be used in EIAs. The present article reviews assays developed with this enzyme, highlights its salient features and brings out an argument in favour of its wide spread use in EIAs. PMID- 10458641 TI - A method for correcting for the variability of inhibitory effects of soluble human interleukin 1 receptor II measured by different ELISAS. AB - Seven ELISAs were developed by using several combinations of anti-human IL-1beta antibodies for detecting interleukin 1beta (IL-1beta) in cell culture supernatants. These ELISAs have different sensitivities in detecting standard preparations of recombinant human IL-1beta (WHO reference standard) compared with conventional preparations of IL-1beta produced by stimulated human peripheral blood mononuclear cells. The observed differences were attributed to differences in epitope specificity of the various monoclonal antibodies used and the heterogeneity of IL-1beta secreted into culture supernatants. The presence of soluble IL-1 receptor type I did not alter the levels of IL-1beta detected by these ELISAs. However, soluble IL-1 receptor type II interfered with the detection of IL-1beta to different degrees in these ELISAs. A method involving standarization by means of separate measurement of the amount of receptor and its inhibitory effect in the IL-1beta ELISA, yields consistent estimates of the correct IL-1beta levels. PMID- 10458642 TI - Angiotensin II and connective tissue: homeostasis and reciprocal regulation. AB - As a concept traditionally applied to integrative organ physiology, homeostasis likewise applies to self-regulated growth and structure of loose, dense and specialized connective tissues. De novo generation and co-induction of signals, either stimulatory or inhibitory to the formation of these tissues, provide for a reciprocal regulation of their composition; angiotensin (Ang) II is a growth stimulator. Components involved in AngII generation and its biological activity, including angiotensin converting enzyme (ACE) and AngII receptors, are expressed by mesenchymal cells responsible for connective tissue turnover. ACE inhibition or AT1 receptor antagonism attenuate the formation of these connective tissues. The concept of circulatory homeostasis, and the endocrine properties of plasma AngII involved in maintaining same, need each be broadened to encompass auto- and paracrine effects of AngII produced within connective tissues, where it contributes to their homeostatic regulation of structure and composition. PMID- 10458643 TI - Acceleration of ulcer healing by cholecystokinin (CCK): role of CCK-A receptors, somatostatin, nitric oxide and sensory nerves. AB - CCK exhibits a potent cytoprotective activity against acute gastric lesions, but its role in ulcer healing has been little examined. In this study we determined whether exogenous CCK or endogenously released CCK by camostate, an inhibitor of luminal proteases, or by the diversion of pancreatico-biliary secretion from the duodenum, could affect ulcer healing. In addition, the effects of antagonism of CCK-A receptors (by loxiglumide, LOX) or CCK-B receptors (by L-365,260), an inhibition of NO-synthase by N(G)-nitro-L-arginine (L-NNA), or sensory denervation by large neurotoxic dose of capsaicin on CCK-induced ulcer healing were examined. Gastric ulcers were produced by serosal application of acetic acid and animals were sacrificed 9 days after ulcer induction. The area of ulcers and blood flow at the ulcer area were determined. Plasma levels of gastrin and CCK and luminal somatostatin were measured by RIA and mucosal biopsy samples were taken for histological evaluation and measurement of DNA synthesis. CCK given s.c. reduced dose dependently the ulcer area; the threshold dose of CCK being 1 nmol/kg and the dose inhibiting this area by 50% being 5 nmol/kg. This healing effect of CCK was accompanied by a significant increase in the GBF at ulcer margin and the rise in luminal NO production, plasma gastrin level and DNA synthesis. Concurrent treatment with LOX, completely abolished the CCK-8-induced acceleration of the ulcer healing and the rise in the GBF at the ulcer margin, whereas L-365,260 remained without any influence. Treatment with camostate or diversion of pancreatic juice that raised plasma CCK level to that observed with administration of CCK-8, also accelerated ulcer healing and this effect was also attenuated by LOX but not by L-365,260. Inhibition of NO-synthase by L-NNA significantly delayed ulcer healing and reversed the CCK-8 induced acceleration of ulcer healing, hyperemia at the ulcer margin and luminal NO release, and these effects were restored by the addition to L-NNA of L-arginine but not D-arginine. Capsaicin denervation attenuated CCK-induced ulcer healing, and the accompanying rise in the GBF at the ulcer margin and decreased plasma gastrin and luminal release of somatostatin when compared to those in rats with intact sensory nerves. Detectable signals for CCK-A and B receptor mRNAs as well as for cNOS mRNA expression were recorded by RT-PCR in the vehicle control gastric mucosa. The expression of CCK-A receptor mRNA and cNOS mRNA was significantly increased in rats treated with CCK-8 and camostate, whereas CCK-B receptor mRNA remained unaffected. We conclude that CCK accelerates ulcer healing by the mechanism involving upregulation of specific CCK-A receptors, enhancement of somatostatin release, stimulation of sensory nerves and hyperemia in the ulcer area, possibly mediated by NO. PMID- 10458644 TI - Cellular localization, expression and regulation of neuropeptide Y in kidneys of hypertensive rats. AB - Neuropeptide Y (NPY) is a key modulator of the autonomic nervous system playing pivotal roles in cardiovascular and neuronal functions. In this study, we assessed the cellular localization and gene expression of NPY in rat kidneys. We also examined the relationship between NPY gene expression and renin in two rat models of hypertension (two-kidney, one-clip renal hypertension (2K1C), and deoxycorticosterone-salt-induced hypertension (DOCA-salt)) characterized by a similar blood pressure elevation. In situ hybridization and immunohistochemistry, using anti-NPY or anti-C-flanking peptide of NPY (CPON) antibodies, showed that NPY transcript and protein were colocalized in the tubules of rat kidneys. During experimental hypertension, NPY mRNA was decreased in both kidneys of the 2K1C animals, but not in the kidney of DOCA-salt rats. In 2K1C rats, renal NPY content was also decreased. The difference in NPY gene expression between 2K1C rats (a high renin model of hypertension) and DOCA-salt rats (a low renin model of hypertension) suggests that circulating angiotensin II plays a role in local renal NPY gene expression and that the elevated blood pressure per se is not the primary factor responsible for the control of NPY gene expression in the kidney. PMID- 10458645 TI - Effects of vasopressin-mastoparan chimeric peptides on insulin release and G protein activity. AB - Two chimeric peptides, consisting of the linear vasopressin receptor V1 antagonist PhAc-D-Tyr(Me)-Phe-Gln-Asn-Arg-Pro-Arg-Tyr, in the N-terminus and mastoparan in the C-terminus connected directly (M375) or via 6-aminohexanoic acid (M391), have been synthesised. At 10 microM concentration, these novel peptides increased insulin secretion from isolated rat pancreatic islet cells 18 26-fold at 3.3 mM glucose and 3.5-5-fold at 16.7 mM glucose. PTX pretreatment of the islets decreased the peptide-induced insulin release. M375 and M391 bind to V1a vasopressin receptors with affinities lower than the unmodified vasopressin antagonist, but with K(D) values of 3.76 nM and 9.02 nM, respectively, both chimeras are high affinity ligands. The GTPase activity and GTPgammaS binding in the presence of these peptides has been characterised in Rin m5F cells. Comparison of the influence of the peptides M375 and M391 on GTPase activity in native and pertussis toxin-treated cells suggests a selective activation of G alpha(i)/G alpha(o) subunits, combined with a suppression of other GTPases, primarily G alpha(s). However, the GTPgammaS binding data show that the peptides retain some of the activating property even in PTX-treated cell membranes. In conclusion, the conjugation of mastoparan with the V1a receptor antagonists produce peptides with properties different from the parent peptides that could be used to elucidate the role of different G proteins in insulin release. PMID- 10458646 TI - Evidence for calcitonin gene-related peptide-mediated ischemic preconditioning in the rat heart. AB - Previous studies have suggested that calcitonin gene-related peptide (CGRP) may play an important role in the mediation of ischemic preconditioning. In the present study, we examined the release of CGRP during ischemic preconditioning and the effect of preconditioning frequency on this effect in the isolated rat heart. Thirty minutes of global ischemia and 40 min of reperfusion caused a significant cardiac dysfunction and an increase in the release of creatine kinase (CK) during reperfusion. Preconditioning with one, two or three cycles of 5-min ischemia and 5-min reperfusion caused a marked improvement of cardiac function and a decrease in the release of CK, and there was no difference in the degree of improvement among groups. The protective effects of ischemic preconditioning were abolished by the CGRP receptor antagonist CGRP(8-37). A single preconditioning cycle induced a significant increase in the release of CGRP in the coronary effluent. In the hearts treated with two or three preconditioning cycles, the level of CGRP was highest in the first cycle, and was gradually decreased with increasing number of cycles of preconditioning. These results suggest that the protective effects of ischemic preconditioning are mediated by endogenous CGRP in the isolated rat heart. PMID- 10458647 TI - Active immunoneutralization of somatostatin in the sheep: effects on gastrointestinal somatostatin expression, storage and secretion. AB - In the absence of somatostatin antagonists, somatostatin antisera administered acutely or animals chronically immunized against somatostatin have been used to define the functions of somatostatin. However, the circulating immunoglobulins from immunized animals may contain substantial quantities of endogenous hormones. This has not been examined for somatostatin. We have measured the amount of free somatostatin bound to circulating immunoglobulins in somatostatin-immunized animals and the effect of this sequestering of the free peptide on somatostatin secretion and gastric somatostatin synthesis and storage. The average concentration of somatostatin bound to the antisera was 6.9 nmol/l, about 1000 fold higher than normal circulating levels. Compared to control animals, there was a doubling of somatostatin mRNA in the fundus and a 4-fold increase in fundic somatostatin peptide. Similar increases were seen in pancreas, but the antrum was not significantly affected providing further evidence of distinct regulatory mechanisms between the antrum and fundus. We suggest that withdrawal of active somatostatin activates a regulatory loop to increase fundic somatostatin biosynthesis and storage. The data support the concept that somatostatin autoregulates its own expression at both the RNA and peptide level. PMID- 10458648 TI - Mast cells contribute to PACAP-induced dermal oedema in mice. AB - In the present study the effect of intradermal PACAP-injection on dermal oedema in mice was investigated and the contribution of mast cells to this response was assessed. The injection of PACAP 1-38 into the ears of C57BL/6 mice evoked a dose dependent response, which, after higher doses of PACAP 1-38, lasted at least 24 h. Histological examination showed significant mast cell degranulation induced by PACAP. Using mast cell-deficient WBB6F1-Kit(W)/Kit(W-v) mice and the congenic mice, we demonstrated that the the early phase (30 min to 6 h) of PACAP-induced ear swelling response was significantly diminished in mast cell-deficient mice, suggesting that mast cell degranulation contributes to this phase of the response. When mast cell-deficient WBB6F1-Kit(W)/Kit(W-v) mice were locally and selectively reconstituted by adoptive mast cell transfer, the dermal oedema was almost equal to that of control animals in the early phase of PACAP injection. These results show that mast cell degranulation contributes to PACAP-induced dermal oedema in mice. PMID- 10458649 TI - Islet neuronal abnormalities associated with impaired insulin secretion in type 2 diabetes in the Chinese hamster. AB - This study examined the relationship between islet neurohormonal characteristics and the defective glucose-stimulated insulin secretion in genetic type 2 diabetic Chinese hamsters. Two different sublines were studied: diabetes-prone CHIG hamsters and control CHIA hamsters. The CHIG hamsters were divided into three subgroups, depending on severity of hyperglycemia. Compared to normoglycemic CHIG hamsters and control CHIA hamsters, severely hyperglycemic CHIG hamsters (glucose > 15 mmol/l) showed marked glucose intolerance during i.p. glucose tolerance test and 75% impairment of glucose-stimulated insulin secretion from isolated islets. Mildly hyperglycemic CHIG animals (glucose 7.2-15 mmol/l) showed only moderate glucose intolerance and a 60% impairment of glucose-stimulated insulin secretion from the islets. Immunostaining for neuropeptide Y and tyrosine hydroxylase (markers for adrenergic nerves) and for vasoactive intestinal peptide (marker for cholinergic nerves) revealed significant reduction in immunostaining of islets in the severely but not in the mildly hyperglycemic animals, compared to control CHIA hamsters. The study therefore provides evidence that in this model of type 2 diabetes in Chinese hamsters, severe hyperglycemia is accompanied not only by marked glucose intolerance and islet dysfunction but also by reduced islet innervation. This suggests that islet neuronal alterations may contribute to islet dysfunction in severe but not in mild diabetes. PMID- 10458650 TI - Allatostatins from the retrocerebral complex and antennal pulsatile organ of the American cockroach: structural elucidation aided by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. AB - The occurrence of allatostatins in retrocerebral complexes and antennal pulsatile organs of the American cockroach, Periplaneta americana, was investigated. Previously, molecular cloning of the P. americana allatostatin gene had predicted 14 peptides of this family [Ding et al., Comparison of the allatostatin neuropeptide precursors in the distantly related cockroaches Periplaneta americana and Diploptera punctata. Eur J Biochem 1997;234:737-746], however, only two forms had been identified by peptide isolation procedures [Weaver et al., Identification of two allatostatins from the CNS of the cockroach Periplaneta americana: novel members of a family of neuropeptide inhibitors of insect juvenile hormone biosynthesis. Comp Biochem Physiol 1994;107(C):119-127]. Using an extract of only 200 corpora cardiaca/corpora allata, we have found that at least 11 allatostatins occur in the retrocerebral complex. These peptides were already separated from other substances of the crude extract in the first HPLC step with heptafluorobutyric acid as organic modifier, and subsequently identified by MALDI-TOF mass spectrometry. Moreover, we have demonstrated the occurrence of nearly all allatostatins, including the cleavage product of Pea-AST 2 (LPVYNFGL-NH2), in antennal pulsatile organs of males and females. Allatostatins are predominant neuropeptides in these organs. Additionally, only two other known peptides could be identified in these organs by mass screening: proctolin and leucomyosuppressin. The function of allatostatins in antennal pulsatile organs remains unclear. We assume a release into the hemolymph via the ampullac, which could act as neurohemal release sites. The method described for the identification of allatostatins is a very fast method for neuropeptide screening in neurohemal tissues. PMID- 10458651 TI - Characterization of rabbit kidney and brain pancreatic polypeptide-binding neuropeptide Y receptors: differences with Y1 and Y2 sites in sensitivity to amiloride derivatives affecting sodium transport. AB - Sites sensitive to human and rat pancreatic polypeptides (hPP and rPP) accounted for more than 30% of the specific binding of [125I](Leu31,Pro34) human peptide YY (LP-PYY) in particulates from rabbit kidney cortex, and about 10% of the specific binding in membranes from rabbit hypothalamus. The binding of [125I]hPP or [125I]rPP showed a high-affinity displacement with either hPP, rPP, LP-PYY, neuropeptide Y or peptide YY (Ki below 50 pM for all), while being quite insensitive to Y2-selective ligands. The PP binding had a high sensitivity to alkali cations and inhibitors of phospholipase C, very similar to that of LP-PYY binding 'masked' by excess cold hPP. However, as different from the Y1-like LP PYY binding, but similar to the binding of the Y2-selective ligand [125I]human peptide YY(3-36) (hPYY(3-36)), the PP binding showed a low sensitivity to guanosine polyphosphates. The PP binding was much more sensitive to N5 substituted amiloride inhibitors of Na+ transport than the binding of LP-PYY, or that of hPYY(3-36). The inhibition of PP binding by N5-substituted amilorides was not enhanced by guanine nucleotides or by phospholipase C blockers. However, pairing of N5-substituted amilorides disproportionately increased the inhibition of hPP binding. Thus, in rabbit kidney or hypothalamus, the high-affinity PP responding sites share some of the basic properties of the Y1 and Y2 sites, but are distinguished from both by a high sensitivity to compounds affecting sodium transport. These PP/NPY receptors could associate with membrane structures involved in the control of ion balance and osmotic responses. PMID- 10458652 TI - Recent advances on neuronal caspases in development and neurodegeneration. AB - In view of a large and growing literature, this overview emphasizes recent advances in neuronal caspases and their role in cell death. To provide historical perspective, morphology and methods are surveyed with emphasis on early studies on interleukin converting enzyme (ICE) as a prototype for identifying zymogen subunits. The unexpected homology of ICE (caspase-1) to Caenorhabditis elegans death gene CED-3 provided early clues linking caspases to programmed cell death, and led later to discovery of bcl-2 proteins (CED-9 homologs) and 'apoptosis associated factors' (Apafs). Availability of substrates, inhibitors, and cDNAs led to identification of up to 16 caspases as a new superfamily of unique cysteine proteinases targeting Asp groups. Those acting as putative death effectors dismantle neurons by catabolism of proteins essential for survival. Caspases degrade amyloid precursor protein (APP), presenilins (PS1, PS2), tau, and huntingtin, raising questions on their role in neurodegeneration. Brain contains 'inhibitors of apoptosis proteins' (IAPs) survivin and NAIP associated also with some neuronal disorders. Apoptotic stress in neurons initiates a chain of events leading to activation of distal caspases by pathways that remain to be fully mapped. Neuronal caspases play multiple roles for initiation and execution of cell death, for morphogenesis, and in non-mitotic neurons for homeostasis. Recent studies focus on cytochrome c as pivotal in mediating conversion of procaspase-9 as a major initiator for apoptosis. Identifying signaling pathways and related events paves the way to design useful therapeutic remedies to prevent neuronal loss in disease or aging. PMID- 10458653 TI - Distribution of plasma membrane Ca2+ pump (PMCA) isoforms in the gerbil brain: effect of ischemia-reperfusion injury. AB - Non-species isoform-specific antibodies against three isoforms of the plasma membrane Ca2+ pump (PMCA) were used for immuno-localization of PMCA by Western blot analysis in membrane preparations isolated from different regions of gerbil brain. All three gene products were detected in the membranes from hippocampus, cerebral cortex and cerebellum. However, they showed a distinct distribution pattern. Two proteins were revealed in the case of PMCA1 with molecular masses 129 and 135 kDa. The antibody against PMCA2 recognized three proteins of about 130-137 kDa. Only one protein was detected with the anti-PMCA3 antibody. Levels of immuno-signal for the PMCA isoforms varied significantly among the different brain regions. The PMCA1 is the most abundant in the cerebro-cortical and hippocampal membrane preparations. The PMCA2 was detected in a lesser amount comparing to PMCA1 and was highest in the membrane preparations from cerebellum and in a slightly lesser amount from cerebral cortex. Anti-PMCA3 antibody stained weakly and was localized in the cerebellar and hippocampal membrane preparations. Transient forebrain ischemia (10 min) and reperfusion (for a prolonged period up to 10 d) leads to a significant decrease of PMCA immuno-signal. This decrease could be ascribed to the loss of PMCA1 signal, especially in hippocampal membrane preparations. PMID- 10458654 TI - Cyclosporin A regulates the levels of cyclophilin A in neuroblastoma cells in culture. AB - Cyclophilin A (CyP-A), a member of a highly conserved family of proteins, immunophilins, is the major intracellular receptor for the immunosuppressive drug, cyclosporin A (CsA). CyP-A is widely expressed in many tissues, but is found in the highest concentration in brain tissues and may perform critical neuronal functions. CsA is a known neurotoxin. Therefore, understanding the regulation of CyP-A levels in nerve cells, particularly by CsA, is important. We have utilized murine neuroblastoma (NB) cells as an experimental model to investigate this issue. Our results show that CsA alone was sufficient to induce morphological differentiation in undifferentiated NB cells and to increase CyP-A levels as determined by immunostaining. However, inducing terminal differentiation by elevating adenosine 3',5'-cyclic monophosphate (cAMP) levels using either 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (RO20-1724), an inhibitor of cyclic nucleotide phosphodiesterase, or prostaglandin E1 (PGE1), a stimulator of adenylate cyclase, was not sufficient to increase CyP-A levels. CsA was required to increase CyP-A levels in both RO20-1724- and PGE1-induced differentiated NB cells. Increases in CyP-A levels, however, occurred without any change in the expression of the CyP-A gene as determined by reverse-transcriptase polymerase-chain reaction analysis using (CyP-A)-specific primers. These results suggest that CsA regulates the level of its own binding protein, CyP-A, in both undifferentiated and cAMP-induced differentiated NB cells in culture. PMID- 10458655 TI - Nervous control of the cerebrovascular system: doubts and facts. AB - Increased function of the central neurons results in increased neuronal metabolism and, as a consequence, increased concentration of metabolic end products (H+, K+, adenosin) results in an increased cerebral blood flow (CBF). There is a general agreement among investigators that products of cerebral tissue metabolism as well as chemical stimuli are key factors that determine the rate of blood flow in the brain. CBF, however, may increase out of proportion to metabolic demands, may increase without significant change in local metabolism, and may increase much faster than the accumulation of the metabolic end-products. Therefore, the 100-year-old metabolic hypothesis of Roy and Sherrington, cannot fully explain the increases of CBF during increased functional activity of the central neurons. The tight coupling of neuronal activity and blood flow in the brain is demonstrated by a large amount of data. Therefore, the likelihood exists that neurogenic stimuli via perivascular nerve endings may act as rapid initiators, to induce a moment-to-moment dynamic adjustment of CBF to the metabolic demands, and further maintenance of these adjusted parameters is ensured by the metabolic and chemical factors. Perivascular nerve endings were identified in the outer smooth muscle layer of the cerebral arteries, arterioles and veins. Their axonterminals contain a large variety of neurotransmitters, often co-localised in synaptic vesicles. Stimulation of the nerves results in a release of transmitters into the narrow neuromuscular synaptic clefts in the cerebrovascular smooth muscle, close to specific receptor sites in the vessel wall. In spite of these facts, however, and in spite of the large number of new experimental evidences, the role of the nervous control of the cerebrovascular system is underestimated both in medical textbooks and in the common medical knowledge since decades. In the last 20 years major advances have been made that make it necessary to revise this false view. The purpose of this review is to facilitate this process at the end of this century, when the importance of the nervous control of the cerebral circulation has been fully appreciated among investigators. PMID- 10458656 TI - Butyric acid mediated induction of enhanced transendothelial resistance in an in vitro model blood-brain barrier system. AB - Previously we reported that the co-culture of non-brain vascular endothelial cells with glioma cells leads to the induction of a more differentiated endothelial cell phenotype which exhibits important properties of the blood-brain barrier (BBB). Recognising the potential for improving the model barrier system with agents known to modify the growth and differentiation of cells in culture we examined the effects of four differentiating agents (butyric acid, dexamethasone, retinoic acid, and dimethyl sulfoxide) on barrier function. Of these agents only butyric acid and dexamethasone resulted in an enhancement (depending on the dose used) of transendothelial electrical resistance (barrier function). The greatest effect was observed with butyric acid in a dose-dependent manner and was slow in onset and only occurred in the endothelial/glial cell co-cultures. These data indicate that butyric acid may be a beneficial agent in optimising conditions necessary for induction of BBB properties in in vitro barrier systems. PMID- 10458657 TI - Tolterodine for overactive bladder: time to onset of action, preferred dosage, and 9-month follow-up. AB - This is a prospective study of 28 patients who had urinary frequency (>8 times/day) and either urgency or urge incontinence (>1 time/day). After a 2-week run-in period (visit 1), the patients were started on tolterodine 1 mg twice a day (bid) (visit 2). They were followed at 4 and 8 weeks (visits 3 and 4). The patients were contacted by telephone 1 week after visit 2. Tolterodine was increased to 2 mg bid if the patient had incomplete improvement at either the initial phone call or during visit 3. Evaluation criteria were daily micturition charts including urinary frequency, nocturia, leakage episodes, average urine volume per day, and average voided volume. Tolterodine was well tolerated without side effects in 20 (80%) of 28 patients. Eight patients (20%) dropped out after enrollment because of side effects in 3, no improvement in 2, and missing visits (>1) in 3. Drug dosage in the 20 patients who tolerated tolterodine was 1 mg bid in 3 and 2 mg bid in 17 (85%). According to micturition charts, urinary frequency, nocturia, and leakage episodes decreased significantly after tolterodine treatment, whereas average urine volume per day and average voided volume did not change significantly. There were no electrocardiographic or biochemical abnormalities due to tolterodine treatment. Mean follow-up was 9.4 months. All 20 patients who tolerated tolterodine continue to take the medication without significant side effects. We conclude that tolterodine is well tolerated and effective for overactive bladders. Two milligrams bid is the dosage preferred by the majority of patients and the onset of action is seen within 1 week of treatment. Long-term compliance and efficacy are excellent, with no dropout in >9 months of follow-up. PMID- 10458658 TI - Combination and multivariate analysis of PSA-based parameters for prostate cancer prediction. AB - The aim of this study was to evaluate the ability of prostate-specific antigen (PSA)-based parameters including PSA density (PSAD), PSAD of the transition zone (PSA-TZ), percent free PSA, PSA velocity, and their combination to enhance the specificity of PSA for prostate cancer detection in men with serum PSA levels between 4 and 10 ng/mL. We evaluated prospectively 559 consecutive men referred for early detection of prostate cancer who had serum PSA levels between 4 and 10 ng/mL. All men underwent prostatic ultrasonography and sextant biopsy with two additional TZ biopsies. In all cases, if first biopsies were negative an additional set of biopsies was obtained within 6 weeks. The ability of PSAD, PSA TZ, PSA velocity, percent free PSA, and their combination to improve the detection of prostate cancer was evaluated by univariate and multivariate analysis as well as receiver operating characteristic (ROC) curves. In this prospective study of 559 patients, 217 had prostate cancer and 342 had histologically confirmed benign prostatic hyperplasia. Multivariate analysis and ROC curves showed that PSA-TZ and percent free PSA (f/t PSA) were the most powerful and highly significant predictors of prostate cancer. Areas under the ROC curve (AUC) for PSA-TZ and percent free PSA were 0.827 and 0.778, respectively (p = .01). Combination of f/t PSA with PSA-TZ (AUC = 88.1%) significantly increased AUC as compared to each of the other parameters alone as well as their combination (p = .02). The next best combinations were PSA-TZ + PSAD, PSA-TZ + PSA, and f/t PSA + PSA. PSA-TZ followed by f/t PSA and PSAD were the most powerful predictors of prostate cancer in referred patients with a serum PSA between 4 and 10 ng/mL. f/t PSA + PSA-TZ was the most effective combination. When volume-independent PSA parameters were taken into consideration, f/t PSA + PSA clearly outperformed the other options. PMID- 10458659 TI - Endourologic treatment of renal pelvic and ureteral transitional cell carcinoma. AB - Endoscopic treatment of patients with upper urinary tract transitional cell carcinoma is recommended in those with tumor in a solitary kidney, bilateral disease, renal dysfunction, and significant intercurrent illness that precludes a major abdominal surgical procedure. Endoscopic management also may be appropriate in selected patients with small, low-grade lesions in the presence of a normal contralateral kidney. Almost all ureteral tumors and some renal collecting system lesions can be managed using rigid and/or flexible ureteroscopy, which is associated with less bleeding and more rapid recovery than a percutaneous approach. However, larger renal malignancies can be managed effectively using percutaneous resection. Six patients with upper tract transitional cell carcinoma underwent endoscopic resection. The antegrade and retrograde surgical techniques are described. With follow-up up to 23 months, local recurrence in two patients was managed successfully by repeat endoscopic resection. No patient has experienced disease progression or developed metastases. PMID- 10458660 TI - Penile arteriography. AB - Vasculogenic impotence can be due to multiple different causes, many of which can be diagnosed by noninvasive studies. Once an arterial etiology for impotence is suspected, penile arteriography may be necessary to define arterial pathology and to plan potential surgical intervention. The quality of arterial imaging is highly dependent on attention to the technical aspects of the examination, which include a thorough understanding of the arterial anatomy of the pelvis. In addition, patient preparation, arterial catheterization techniques, and pharmacological adjuncts can be helpful to improve image quality. The technique of image acquisition also can affect the quality of the images obtained. This article discusses the technical issues involved in penile arteriography. PMID- 10458661 TI - Application of retrograde cerebral perfusion and moderate systemic hypothermic circulatory arrest for cavoatrial tumor resection. AB - Renal tumors invading the inferior vena cava have proved to be surgically challenging. For suprahepatic and right atrial involvement, deep hypothermic circulatory arrest (HCA) has been the favored procedure. Retrograde cerebral perfusion (RCP) was combined with moderate HCA in an effort to improve cerebral protection and avoid neurological sequelae. Six patients (mean age 64.7 years) who were operated on using this technique underwent a retrospective medical record analysis. The six patients achieved a mean cardiopulmonary bypass, HCA, and RCP time of 2 hours 34 minutes, 26.5 minutes, and 22 minutes, respectively. There were no focal cerebral defects or mortalities. Transient cerebral events were avoided compared to previous patients subjected to deeper hypothermia. All patients are living with no evidence of cancer 16-30 months after surgery. The addition of RCP to HCA for resection of supradiaphragmatic renal carcinoma enhances cerebral protection and allows time for a more efficacious operative procedure. PMID- 10458662 TI - Multicomponent penile prosthesis implantation under regional anesthesia. AB - Fourteen consecutive patients underwent placement of a multicomponent penile prosthesis under regional anesthesia using a pudendal nerve block. Results demonstrated the need for minimal administration of intraoperative narcotics, no major fluctuations in the blood pressure or pulse rate during intraoperative monitoring, and only 1 of 14 patients was converted to general anesthesia. Postoperatively, our results show that there was no urinary retention or cardiac side effects, adequate postoperative analgesia was obtained, and 13 of the 14 patients were discharged within 24 hours. This method was safe, effective, easily performed, and does not carry the associated morbidity of general or spinal anesthesia. PMID- 10458663 TI - Scrotal (Bianchi) approach to patent processus vaginalis in children. AB - Ligation of an indirect inguinal hernia or communicating hydrocele is the most frequent operation performed on children by pediatric surgeons and urologists. We describe our experience with the scrotal (Bianchi) approach to these problems rather than the traditional inguinal approach. PMID- 10458664 TI - An alternative approach to bladder autoaugmentation. AB - This article describes in detail the alternative technique of bladder autoaugmentation. The authors applied a different autoaugmentation method to reliably create a diffuse circular bulge or diverticulum by incising detrusor muscle while leaving bladder mucosa intact. The results were assessed by urodynamic, radiological, and histopathological methods. PMID- 10458665 TI - Simple technique of ligation and division of dorsal vein complex during radical retropubic prostatectomy. AB - A simple technique for ligation and division of the dorsal vein complex during radical retropubic prostatectomy is described. The dorsal vein complex and anterior wall of the membranous urethra were collectively ligated with a "theta theta" suture using 1-0 catgut and then divided during radical retropubic prostatectomy. This technique prevents dislodgment of the catgut during the procedure, enables maximal preservation of the striated urethral sphincter, and guarantees a lower risk of postoperative urinary incontinence. PMID- 10458666 TI - Modified method for closure of transverse pyelotomy incision in Gil-Vernet pyelolithotomy operation. AB - A technique is described for effective closure of the transverse pyelotomy incision used to initiate the Gil-Vernet extended pyelolithotomy operation. Long preplaced sutures of 5-0 Vicryl are made on the intact pelvis across the proposed incision line. After stone removal, the preplaced sutures are tied to approximate the edges of the pyelotomy incision. This simple method helps reduce the chance of pelvic tear, which is a common occurrence when the conventional method of closure is used. PMID- 10458667 TI - Bilateral ureteritis cystica with unilateral ureteropelvic junction obstruction. AB - Ureteritis cystica is a rare, benign, proliferative disorder characterized by multiple ureteral cysts and multiple filling defects noted on contrast ureteral imaging. A unique case of bilateral ureteritis cystica coincidental with chronic, congenital, unilateral ureteropelvic junction obstruction presenting with microscopic hematuria and lower urinary tract symptoms is described. The characteristic presentation as well as the diagnostic radiographic, ureteroscopic, and histologic features of pyeloureteritis cystica are reviewed. PMID- 10458668 TI - Unilateral obstruction of the vas deferens diagnosed by seminal vesicle aspiration. AB - Causes of vasal obstruction include vasectomy, inguinal surgery, scrotal surgery, and congenital anomalies. The incidence of unilateral obstruction in various clinical situations is unknown because sperm from the contralateral testicle usually is present in the ejaculate. Vasography is the standard technique used to diagnose a unilateral vas deferens obstruction. We used the technique of seminal vesicle aspiration to accurately diagnose unilateral vas deferens obstruction in a man who had a previous inguinal hernia repaired with mesh. Surgical exploration confirmed the impression of vasal obstruction, and successful vasovasostomy was performed. We believe that seminal vesicle aspiration may be helpful in the diagnosis of unilateral vas deferens obstruction and has potential benefits over vasography. PMID- 10458669 TI - Genital incarceration with metal rings: their safe removal. AB - Strangulation of the genitalia with constricting metal bands presents a difficult problem with removal. We report the easy and safe removal by cutting the metal bands in two places using a hand-held Dremel Moto-Tool [corrected] with a metal cutting disk. PMID- 10458670 TI - Case No. 1. Computed tomography (CT) of a 70-year-old male presenting with sepsis and right renal mass. PMID- 10458671 TI - Case No. 2. A 5-year-old male with undescended testes. PMID- 10458672 TI - Case No. 3. Newborn with congenital anomaly. PMID- 10458673 TI - Case No. 4. Newborn with prenatal hydronephrosis: what further studies are necessary? PMID- 10458674 TI - Sphincter preservation for distal rectal cancer: paradise lost? PMID- 10458675 TI - Sentinel lymphadenectomy for melanoma--is it a substitute for elective lymphadenectomy? PMID- 10458676 TI - Refining the optimal technical approach for sentinel node localization in breast cancer patients. PMID- 10458677 TI - The impact of local regional therapy on breast cancer survival. PMID- 10458678 TI - An expanded diagnostic role for core biopsy: assessment of the proportion of the duct carcinoma in situ component in invasive carcinomas. PMID- 10458679 TI - Surgical oncology: a specialty in evolution. AB - Surgical oncology has established its role in the multidisciplinary care of the cancer patient. Surgical oncology fellowships are organized to teach multimodality treatment. The typical fellow has completed 6 years of general surgery residency and 1 year in the laboratory with the resultant eight publications. Data compiled from the review of two Society of Surgical Oncology approved fellowship programs, the Surgical Residency Review Committee and the American Board of Surgery, indicate that the majority of fellows join academic faculties and enhance the training of general surgeons, who, in turn, have the major responsibility for oncologic care of the population at large. PMID- 10458680 TI - Sphincter-sparing treatment for distal rectal adenocarcinoma. AB - BACKGROUND: Studies suggest that the anal sphincter can be preserved in some patients with distal rectal adenocarcinoma (DRA), but this has not been validated in any prospective multi-institutional trial. METHODS: To test the hypothesis that the anal sphincter can be preserved in some patients with DRA, the Cancer and Leukemia Group B and collaborators reviewed 177 patients who had T1/T2 adenocarcinomas < or = 4 cm in diameter, which encompassed < or = 40% of bowel wall circumference, and were < or = 10 cm from the dentate line. Of the 177 patients, 59 patients who were eligible for the study had T1 adenocarcinomas and received no further treatment; 51 eligible T2 patients received external beam irradiation (5400 cGY/30 fractions 5 days/week) and 5-fluorouracil (500 mg/m2 IV d1-3, d29-31) after local excision. RESULTS: At 48 months median follow-up, 6 year survival and failure-free survival rates of the eligible patients are 85% and 78% respectively. Three patients died of unrelated disease. Two patients were treated for second primary colorectal tumors; both remain disease free (NED). Another eight patients died of disease, four with distant recurrence only. One T1 patient is alive with distant disease. Two T1 and seven T2 patients experienced isolated local recurrences; all underwent salvage abdominoperineal resection (APR). After APR, one T1 and four of seven T2 patients were NED at the time of last visit (2-7 years). One T1 patient died of local and distant disease. Three of seven T2 patients died with distant disease. CONCLUSIONS: We conclude that sphincter preservation can be achieved with excellent cancer control without initial sacrifice of anal function in most patients. After local recurrence, salvage resection appears effective, but longer follow-up time of local and distant disease-free survival is advised before extrapolation to patients with T3 primaries. PMID- 10458681 TI - Efficacy of lymphatic mapping, sentinel lymphadenectomy, and selective complete lymph node dissection as a therapeutic procedure for early-stage melanoma. AB - BACKGROUND: Lymphatic mapping, sentinel lymphadenectomy, and selective complete lymph node dissection (LM/SL/SCLND) is an increasingly popular alternative to elective lymphadenectomy (ELND) for patients with early-stage melanoma. Although several reports have demonstrated the accuracy of the LM/SL technique, there are no data on its therapeutic value. METHODS: We performed a matched-pair statistical analysis of 534 patients with clinical stage I melanoma; one half of the patients were treated with LM/SL and the other half were treated with ELND. Patients in the two treatment groups were matched for age (54% were < or =50 years of age), gender (63% were male patients), site of the primary melanoma (49% were on the extremities, 36% on the trunk, and 15% on the head and neck), and thickness of the primary melanoma (7% were < 0.75 mm, 42% between 0.75 and 1.5 mm, 43% between 1.51 and 4.0 mm, and 8% > 4 mm). Patients in the LM/SL group underwent complete regional lymphadenectomy (SCLND) only if the LM/SL specimen contained metastatic melanoma. RESULTS: The overall incidences of nodal metastases were no different (P = .18) between LM/SL (15.7%) and ELND (12%) groups, but the incidence of occult nodal disease was significantly (P = .025) higher among patients with intermediate-thickness (1.51-4.0-mm) primary tumors who underwent LM/SL (23.7%) instead of ELND (12.2%). Survival data were compared by the log-rank score test. LM/SL/SCLND and ELND resulted in equivalent 5-year rates of disease-free survival (79 +/- 3.3% and 84 +/- 2.2%, respectively; P = .25) and overall survival (88 +/- 3.0% and 86 +/- 2.1%, respectively; P = .98). The LM/SL and ELND groups also exhibited similar incidences of same-basin recurrences (4.8% vs. 2.1%, P = .10, respectively) and in-transit metastases (2.6% vs. 3.8%, P = .48) after tumor-negative dissections. Patients who underwent ELND showed a higher incidence of distant recurrences (8.9% vs. 4.0%, P = .03), but this may be related to the longer follow-up period for these patients (median, 169 months), compared with the LM/SL-treated patients (45 months). Among patients with tumor-positive nodal dissections, the 5-year overall survival rates were higher, and approached significance (P = .077) for patients treated by LM/SL/SCLND (64 +/- 12%) compared with ELND (45 +/- 10%). CONCLUSIONS: These findings suggest that LM/SL/SCLND is therapeutically equivalent to ELND but may be more effective for identifying nodal metastases in patients with intermediate thickness primary tumors. PMID- 10458682 TI - Intradermal radiocolloid and intraparenchymal blue dye injection optimize sentinel node identification in breast cancer patients. AB - BACKGROUND: Radiotracer and blue dye mapping of sentinel lymph nodes (SLN) have been advocated as accurate methods to stage the clinically negative axilla in breast cancer patients. The technical aspects of SLN biopsy are not fully characterized. In this study we compare the results of intraparenchymal (IP) and intradermal (ID) injection of Tc-99m sulfur colloid, to establish an optimal method for SLN localization. METHODS: 200 consecutive patients had SLN biopsy performed by a single surgeon. Of these, 100 (Group I) had IP injection and 100 (Group II) had ID injection of Tc-99m sulfur colloid. All patients had IP injection of blue dye as well. Endpoints included (1) successful SLN localization by lymphoscintigraphy, (2) successful SLN localization at surgery, and (3) blue dye-isotope concordance (uptake of dye and isotope by the same SLN). RESULTS: Isotope SLN localization was successful in 78% of Group I and 97% of group II patients (P < .001). When isotope was combined with blue dye, SLN were found in 92% of group I and 100% of Group II (P < .01). In cases where both dye and isotope were found in the axilla, dye mapped the same SLN as radiotracer in 97% of Group I and 95% of Group II patients. CONCLUSIONS: The dermal and parenchymal lymphatics of the breast drain to the same SLN in most patients. Because ID injection is easier to perform and more effective, this technique may simplify and optimize SLN localization. PMID- 10458683 TI - Does local therapy affect survival rates in breast cancer? AB - BACKGROUND: The goal of this study was to challenge the hypothesis that local recurrence of breast cancer does not affect survival rates, by determining whether survival rates differ for conservative and radical surgical policies. METHODS: This study used prospective long-term follow-up monitoring of two contemporaneous groups of patients, within a single unit, who were treated identically except for the one variable of local treatment policy, i.e., conservative or radical. A total of 451 patients with operable breast cancer were chosen from 567 consecutive patients with breast cancer who were treated between 1970 and 1979 in the University Department of Surgery. The rate of survival 132 months after treatment was used as an outcome measure. RESULTS: Two hundred forty one patients were treated using a conservative approach and 210 were treated using a radical approach. At 132 months, the survival rate (58% vs. 42%) and median survival time (> 132 vs. 100 months) were significantly improved for the radically treated group (P < .01). The treatment groups were comparable in terms of age, menopausal status, tumor size, histologic grading, and Nottingham Prognostic Index values. The advantage of the radical policy persisted when examined in relation to each of these prognostic factors. CONCLUSIONS: Use of radical local treatment yielded a highly significant survival benefit (comparable to that obtained with adjuvant therapy), compared with a conservative approach. This was related to a reduced locoregional recurrence rate and provides evidence that local therapy influences long-term outcomes for patients with breast cancer. High-quality locoregional control should be emphasized, as is systemic therapy, in management policies. Assessment of surgical techniques, particularly in relation to locoregional recurrence rates, should be included in all studies in which surgery is a component of therapy. PMID- 10458684 TI - Accurate prediction of the amount of in situ tumor in palpable breast cancers by core needle biopsy: implications for neoadjuvant therapy. AB - BACKGROUND: Neoadjuvant chemotherapy facilitates breast conservation in stage II breast cancer patients, whose primary tumors are assumed to be invasive because they are palpable. However, chemotherapy may not be indicated in the minority of patients whose clinically T2 tumors are completely or predominantly in situ. Almost all previous studies of core needle biopsy in breast cancer have been concerned with nonpalpable, mammographically detected tumors, and none have evaluated its ability to quantitatively determine the amounts of in situ and invasive disease. METHODS: From September, 1992 to December, 1997, core needle biopsy was performed on all patients presenting to the Kings County Hospital Breast Clinic with palpable breast masses. Carcinoma was present in both core needle biopsy samples and surgical specimens subsequently obtained from 95 of 99 patients. Each specimen was evaluated for tumor type, histologic grade, and the amounts of in situ and invasive carcinoma it contained, and the results from surgical and core needle biopsy specimens from the same patients were then compared. RESULTS: The surgical specimens of 14 patients had completely or predominantly in situ disease. Completely or predominantly invasive disease was present in 67 specimens, and the remaining 14 had significant amounts of both. The high level of agreement between the amounts of in situ and invasive disease in core needle biopsy and surgical specimens is indicated by Pearson and intraclass correlation coefficients of 0.91 (P < .001 and < .00001, respectively). Tumor type was correctly predicted by core needle biopsy in each case. Variables among these patients, including primary tumor size, interval between biopsy and surgery, or administration of neoadjuvant systemic therapy, did not alter agreement between core needle biopsy and surgical specimens. CONCLUSIONS: Core needle biopsy can identify palpable breast tumors that are predominantly or completely in situ, and, thus, avoid unnecessary neoadjuvant chemotherapy. It also can demonstrate that a tumor is predominantly invasive, but cannot rule out small invasive foci. For that purpose, complete surgical excision of the tumor is required. PMID- 10458685 TI - Recent advances in melanoma staging and therapy. AB - BACKGROUND: Recent advances in the staging and treatment of melanoma were reviewed. METHODS: A literature-based review was performed. RESULTS: The current American Joint Committee on Cancer (AJCC) Staging system for melanoma has several drawbacks. Proposed changes in the staging system to take into account simplified tumor thickness categories, tumor ulceration, and the number (rather than size) of nodal metastases will allow stage groups with more uniform prognosis. The widespread application of sentinel lymph node biopsy for nodal staging allows accurate nodal staging with minimal morbidity. Reverse transcriptase-polymerase chain reaction (RT-PCR) is a very sensitive molecular staging test that may prove useful for identifying early metastatic disease. There is finally an effective adjuvant therapy for melanoma--interferon alfa-2b. Other adjuvant therapies, including melanoma vaccines, may provide effective and less toxic alternatives. New immunotherapy and gene therapy strategies are under investigation. CONCLUSIONS: Ongoing and future adjuvant therapy trials will benefit from improved melanoma staging by accrual of homogeneous groups of patients. New approaches for adjuvant therapy await completion of clinical trials. Innovative new therapies offer hope for patients with advanced disease. PMID- 10458686 TI - Thrombocytopenia after isolated limb or hepatic perfusions with melphalan: the risk of heparin-induced thrombocytopenia. AB - BACKGROUND: Three cases of heparin-induced thrombocytopenia (HIT) were observed in patients undergoing isolated limb perfusion (ILP) with melphalan. This occurrence prompted the discontinuation of prophylactic postoperative heparin in ILP patients and its avoidance in patients undergoing isolated hepatic perfusion (IHP). The need to reassess these decisions led to a review of thrombocytopenia in both patient populations. METHODS: Records of all patients treated with ILP or IHP at our institution from July 1992 through November 1996, were reviewed. Nine IHP patients were tested prospectively for heparin-related antibodies using serum samples obtained perioperatively and during the second postoperative week. RESULTS: Thrombocytopenia (< 100,000 platelets/microL) developed postoperatively in 30% of 131 ILP patients and in 77% of 56 IHP patients. No cases of HIT were identified other than the three who had been previously diagnosed. The prevalence of HIT in heparinized ILP patients was 2.8% (3/108). All nine IHP patients developed heparin-related antibodies postoperatively. CONCLUSIONS: Because the prevalence of HIT following ILP is in the range observed in other clinical settings, postoperative heparin prophylaxis is an option. However, it probably should be limited to the first week, and daily platelet counts should be reviewed for a pattern of thrombocytopenia consistent with HIT. The prevalence of heparin related antibodies after IHP is so high that prophylactic heparin should be avoided in this setting. PMID- 10458688 TI - Expression of p53 and RB proteins in squamous cell carcinoma of the esophagus: their relationship with clinicopathologic characteristics. AB - BACKGROUND: Cancer of the esophagus is one of the most malignant tumors and has a poor prognosis. The p53 and retinoblastoma (RB) genes are involved in the regulation of cell population by suppressing cell proliferative activity. Our goal was to clarify whether expression of p53 and RB genes could be prognostic factors in squamous cell carcinoma of the esophagus. METHODS: Tumor samples taken from 73 patients undergoing subtotal esophagectomy were immunohistochemically stained for the p53 and RB genes. An image analyzer was used for quantitative assessment of the staining, and clinicopathologic characteristics of those patients were investigated. RESULTS: Patients in whom p53 expression was high had greater tumor diameter, deeper tumor invasion, and worse prognosis compared with patients in whom p53 expression was low. Patients in whom RB expression was low had a higher incidence of lymph node metastasis and more advanced disease than did those in whom RB expression was high. The combination of p53 and RB expression revealed that the cases with high p53 and low RB expression had significantly worse survival rates and deeper tumor invasion compared with other groups. In various clinicopathologic parameters, (e.g., age, sex, tumor-diameter, tumor type, location, differentiation, TNM classification, TNM stage) tumor type, tumor size, depth of invasion, lymph node involvement, distant metastasis, and combined p53 and RB expression showed significant differences in survival by univariate analysis. Among those six variables, only lymph node involvement showed an independent prognostic factor for survival (P = .0055) by multivariate analysis. CONCLUSIONS: The combination of p53 and RB expression is not a prognostic indicator in the surgical treatment of esophageal cancer. PMID- 10458687 TI - p21Waf1/Cip1 expression is a prognostic marker in curatively resected esophageal squamous cell carcinoma, but not p27Kip1, p53, or Rb. AB - BACKGROUND: p21Waf1/Cip1 (p21), p27Kip1 (p27), p53, and Rb play critical roles in cell cycle regulation and may influence the clinical behavior of tumors. We examined whether their expression is useful to predict survival of patients with esophageal squamous cell carcinoma (ESC). METHODS: Expression of p21, p27, p53, and Rb was studied by the immunohistochemical method in specimens from 62 patients with curatively resected ESC tumors and scored by a computerized image analysis system. RESULTS: The median expression scores of p21, p27, p53, and Rb (14, 12, 27, and 50, respectively) were used as cut-off points to define low and high expression groups for each protein. The 5-year survival rate for the high p21 expression group was 68%; that for the low expression group was 31% (P = .0062). p27, p53, and Rb were not correlated with overall survival. When patients were categorized into four groups based on p21 expression level and lymph node involvement (pN), the survival curves were significantly different (P = .0017). Thus, patients without lymph node involvement but with low p21 expression had survival similar to that of patients with lymph node involvement and high p21 expression. Multivariate analysis showed that age (P = .0102), lymph node involvement (P = .0076), and p21 (P = .0276) were independent prognostic factors. CONCLUSIONS: Expression of p21 is an independent prognostic factor in curatively resected ESC. Definition of new subgroups of patients based on p21 expression may help to enhance the stratification of stage. PMID- 10458689 TI - Dual roles of peptic ulcer in the carcinogenesis or extension of early gastric cancer. AB - BACKGROUND: Early gastric cancer (EGC) often coexists with peptic ulcer. In this study we investigated the roles of peptic ulcer in the carcinogenesis and extension of gastric cancer. METHODS: The clinicopathological characteristics of EGC and peptic ulcer and their relationship, as well as that of the background intestinal metaplasia, were compared among the following three groups: patients with peptic ulcer only inside the EGC (Contained group, 53 patients); patients with peptic ulcer only outside the EGC (Separate group, 26 patients); and patients of EGC with no peptic ulcer (Absent group, 43 patients). RESULTS: In the Separate group, a male preponderance was observed (P = .006), and all EGCs developed in the middle or lower third of the stomach (P = .06). Most of the EGCs were an intestinal type of cancer with severe background intestinal metaplasia. Topographically, 88% of the peptic ulcers in the Separate group developed proximal to the EGC. On the other hand, in the Contained group, most EGCs developed in the middle third of the stomach with an intestinal/diffuse type ratio of 1:1. Peptic ulcers inside the EGC were significantly more shallow than those that developed outside the EGC (P = 0.008). Although the incidences of submucosal cancer were nearly the same among the three groups, the maximum cancer diameter tended to be increased in the Contained group compared to that in the Absent group, and the incidence of lymph node involvement tended to be higher in the Contained group (8%) as compared with the other two groups (4%-5%). CONCLUSIONS: These results suggest that peptic ulcer outside the EGC contributes to the development of the intestinal type of EGC, with the background of more severe intestinal metaplasia during the peptic ulcer healing processes, whereas peptic ulcer inside the EGC develops secondary to EGC and favors cancer extension and metastasis. Peptic ulcer associated with EGC can be considered to exert different biological roles in the carcinogenesis or extension of ECG according to the location of the peptic ulcer. PMID- 10458690 TI - Functional down-regulation of beta1 and beta2 integrins of lamina propria lymphocytes (LPL) and tumor-infiltrating lymphocytes (TIL) in colorectal cancer patients. AB - Integrins play an important role in various lymphocyte functions. In this study, we isolated lamina propria lymphocytes (LPL) and tumor-infiltrating lymphocytes (TIL) from normal and malignant tissues in patients with colorectal cancer, and examined the expression of beta1 and beta2 integrins on these lymphocytes quantitatively with two-color flow cytometry. Both LPL and TIL expressed a lower level of common beta1 chain (CD29) in CD4 and CD8 subpopulations than did peripheral blood lymphocytes (PBL). Among the associated alpha chains, the expression levels of alpha1 (CD49a) and alpha2 (CD49b) were slightly higher, whereas those of alpha4 (CD49d) and alpha6 (CD49f) were markedly reduced in LPL and TIL. No significant differences were observed in expressions of any alpha1 integrin chains between these two lymphocytes populations. Similarly, both alphaL (CD11a) and beta2 (CD18) were down-regulated in TIL and LPL with CD8+ cytotoxic phenotype, but not in those with CD4+ phenotype. CD8+ TIL expressed a slightly but significantly higher level of alphaLbeta2 than did CD8+ LPL. CD8+ LPL and CD8+ TIL consistently showed significantly decreased binding to purified ICAM-1, VCAM-1 and HT29 colon cancer cells as compared with CD8+ PBL. Although CD8+ TIL showed a slightly higher level of adhesion to these substrates than did CD8+ LPL, the level was much lower than that in PBL. The expression pattern and functional down-regulation of these integrins may be one of the reasons why TIL cannot eradicate the cancer cells in colorectal cancer. PMID- 10458691 TI - Society of Surgical Oncology: statement on genetic testing for cancer susceptibilty. Committee on Issues and Governmental Affairs of the Society of Surgical Oncology. PMID- 10458692 TI - Assessment of treatment for patients with primary ductal carcinoma in situ in the breast. PMID- 10458693 TI - Follow-up after treatment of primary breast cancer. PMID- 10458694 TI - Use of touch preps for intraoperative diagnosis of sentinel lymph node metastases in breast cancer. PMID- 10458695 TI - Will the true sentinel node please stand? PMID- 10458697 TI - Letters to the editor PMID- 10458696 TI - Erythema ab igne in a child with atopic eczema. PMID- 10458698 TI - A genetic marker associated with non-response to recombinant hepatitis B vaccine by indigenous Australian children. PMID- 10458699 TI - Uses of the stethoscope. PMID- 10458700 TI - Long-term cost-effectiveness of low molecular weight heparin versus unfractionated heparin for the prophylaxis of venous thromboembolism in elective hip replacement. AB - BACKGROUND AND OBJECTIVE: Either low molecular weight heparin (LMWH) or unfractionated heparin (UH) may be used for the prophylaxis of post-operative venous thromboembolic disease (VTD) in elective hip replacement. This study was aimed at assessing the cost-effectiveness of LMWH over UH from the society perspective, which considers all the outcomes occurring in the life-long time horizon. DESIGN AND METHODS: A decision tree modeled the clinical outcomes and resources used in consequence of restricted (2 weeks) and extended (4 weeks) prophylaxis of VTD with LMWH or UH. RESULTS: In the studied population, that of 67 year-old patients, restricted prophylaxis with LMWH saved 25 quality-adjusted days and $75 over UH. Extended prophylaxis provided a small additional benefit with additional cost savings. The incremental outcomes of the model proved independent of most parameters. INTERPRETATION AND CONCLUSIONS: We conclude that LMWH has considerable advantages over UH in the prophylaxis of VTD following elective hip replacement, and should be recommended in clinical practice. PMID- 10458701 TI - Characterization of the reactivity, regioselectivity, and stereoselectivity of the reactions of butadiene monoxide with valinamide and the N-terminal valine of mouse and rat hemoglobin. AB - Occupational exposure to 1,3-butadiene (BD) has been monitored by measuring the level of hemoglobin N-terminal valine adduct formation with the primary reactive metabolite, butadiene monoxide (BMO). However, mechanistic details concerning the relative reactivity, regioselectivity, and stereospecificity of BMO with the N terminal valine of hemoglobin are lacking. In the studies presented here, L valinamide was used as a model for the N-terminal valine of hemoglobin to compare the nucleophilic reactivity, regioselectivity, and stereoselectivity of the reaction both in aqueous solution and within a protein microenvironment. Four products produced by the reaction of L-valinamide with racemic BMO (two pairs of diastereomers produced by reactions at C-1 and C-2 of the epoxide moiety) were synthesized, purified, and characterized by (1)H NMR and GC/MS. These four reaction products were used as analytical standards for kinetic studies of the reaction of valinamide with BMO at physiological pH (7.4) and temperature (37 degrees C). The results show that the adducts formed by reaction at C-2 were formed at a ratio of approximately 2:1 compared to the adducts formed by reaction at C-1. The stereoisomers of each respective regioisomer were produced with similar rates of formation. The reaction of BMO with the N-terminal valine of hemoglobin was also studied in vitro using intact erythrocytes from Sprague Dawley rats and B6C3F1 mice. After cleavage of the N-modified valine by the N alkyl Edman degradation procedure using pentafluorophenylisothiocyanate (PFPITC), a novel procedure was developed that allowed GC/MS detection and quantitation of the four expected products by silylation of the PFPTH-valine-BMO derivatives. The hemoglobin results contrast with the valinamide results in that the reaction of BMO with the N-terminal valine residue in both rat and mouse hemoglobin produced mostly C-1 adducts. The rates obtained with rat hemoglobin were much slower than the rates obtained with mouse hemoglobin or with valinamide. These results, and the finding that the reaction with rat hemoglobin produced a higher ratio of C1:C2 adducts in comparison with the reaction with mouse hemoglobin, indicate the importance of measuring all four adducts when comparing the relative rates of adduct formation both with model compounds and among different species. PMID- 10458702 TI - 2,2',3,3',6,6'-hexachlorobiphenyl hydroxylation by active site mutants of cytochrome P450 2B1 and 2B11. AB - The structural basis of species differences in cytochrome P450 2B-mediated hydroxylation of 2,2',3,3',6,6'-hexachlorobiphenyl (236HCB) was evaluated by using 14 site-directed mutants of cytochrome P450 2B1 and three point mutants of 2B11 expressed in Escherichia coli. To facilitate metabolite identification, seven possible products, including three hydroxylated and four dihydroxylated hexachlorobiphenyls, were synthesized by direct functionalization of precursors and Ullmann and crossed Ullmann reactions. HPLC and GC/MS analysis and comparison with authentic standards revealed that 2B1, 2B11, and all their mutants produced 4, 5-dihydroxy-236HCB and 5-hydroxy-236HCB, while 2B11 L363V and 2B1 I114V mutants also catalyzed hydroxylation at the 4-position. The amount of products formed by 2B1 mutants I114V, F206L, L209A, T302S, V363A, V363L, V367A, I477A, I477L, G478S, I480A, and I480L was smaller than that of the wild type. I477V exhibited unaltered 236HCB metabolism, and I480V produced twice as much dihydroxy product as the wild type. For 2B11, substitution of Val-114 or Asp-290 with Ile decreased the product yields. Replacement of Leu-363 with Val dramatically altered the profile of 236HCB metabolites. In addition to an increase in the overall level of hydroxylation, the mutant mainly catalyzed hydroxylation at the 4-position. Incubation of P450 2B1 with 5-hydroxy-236HCB produced 4,5-dihydroxy 236HCB, which indicates that 4,5-dihydroxy-236HCB may be formed by a direct hydroxylation of 5-hydroxy-236HCB. The findings from this study demonstrate the importance of residues 114, 206, 209, 302, 363, 367, 477, 478, and 480 in 2B1 and 114, 290, and 363 in 2B11 for 236HCB metabolism. PMID- 10458703 TI - Cytotoxicity and DNA interaction of the enantiomers of 6-amino-3-(chloromethyl)-1 [(5,6,7-trimethoxyindol-2-yl)carbonyl]indo- line (amino-seco-CI-TMI). AB - The enantiomers of the previously reported racemic 6-amino-3-(chloromethyl)-1 [(5,6,7-trimethoxyindol-2-yl)carbonyl] indoline (amino-seco-CI-TMI) were prepared via resolution of a precursor by chiral HPLC. The only detectable product isolated from reaction of the racemic compound with calf thymus DNA, followed by thermal cleavage, was shown by mass spectrometry and two-dimensional NMR spectroscopy to be the adenine N3 adduct. Polyacrylamide gel electrophoresis assays with the racemate and with each enantiomer also showed adenine to be the only site of alkylation. While the racemic amino compound exhibited sequence selectivity identical to that of the previously characterized phenol analogue, the enantiomers exhibited distinctly different sequence selectivities, allowing the (+) enantiomer to be assigned the "natural" S configuration. The (+)-(S) enantiomer is 3-fold more cytotoxic than the (-)-(R) enantiomer (IC(50) values of 240 and 700 nM, respectively, in AA8 cells, after exposure for 4 h). PMID- 10458704 TI - Site-specific synthesis of aflatoxin B(1) adducts within an oligodeoxyribonucleotide containing the human p53 codon 249 sequence. AB - This work describes the preparation of the cationic trans-8, 9-dihydro-8-(N7 guanyl)-9-hydroxyaflatoxin B(1) ((AFB)G) adducts at the positions corresponding to G(746) or G(747), within the oligodeoxyribonucleotide d(GGAGGCCT) containing the codon 249 sequence (underlined) of the p53 gene, using DNA triplexes to target adduction at the desired site. This approach enabled the successful preparation and purification of sufficient quantities of d(GGAG(AFB)GCCT) for NMR structural studies, using only standard phosphoramidites. The presence of multiple guanines in this oligodeoxynucleotide precluded the direct reaction of d(GGAGGCCT). d(AGGCCTCC) with aflatoxin epoxide as a method for producing large quantities of site-specific adducts for physical studies. Of the multiple potential alkylation sites at guanine N7 in d(GGAGGCCT). d(AGGCCTCC), it was found that sites G(2) and G(5) exhibited approximately equal reactivity with aflatoxin B(1)-exo-8,9-epoxide; the reactivity at site G(4) was reduced by approximately a factor of 2 as compared to that at G(2) or G(5). To successfully prepare the site-specific adducts, the p53 oligodeoxyribonucleotide was annealed with either the blocking strand d(CTCCATTTTCCT) or d(CCTCCATTTTCCTC) to form the corresponding partial triplexes which targeted AFB(1) adduction either to G(4) or to G(5). Piperidine cleavage, followed by heating, confirmed that in each instance, the product corresponded to the lone guanine not protected from adduction by the partial DNA triplex. The adducted oligodeoxyribonucleotides were examined with regard to purity by capillary electrophoresis. The primary advantage of this modified triple helix methodology is that it requires only standard phosphoramidites; thus, it is applicable to large-scale preparations that are necessary for NMR structural studies or other physical measurements. PMID- 10458705 TI - Alkylation of human serum albumin by sulfur mustard in vitro and in vivo: mass spectrometric analysis of a cysteine adduct as a sensitive biomarker of exposure. AB - To develop a mass spectrometric assay for the detection of sulfur mustard adducts with human serum albumin, the following steps were performed: quantitation of the binding of the agent to the protein by using [(14)C]sulfur mustard and analysis of acidic and tryptic digests of albumin from blood after exposure to sulfur mustard for identification of alkylation sites in the protein. The T5 fragment containing an alkylated cysteine could be detected in the tryptic digest with micro-LC/tandem MS analysis. Attempts to decrease the detection limit for in vitro exposure of human blood by analysis of the alkylated T5 fragment were not successful. After Pronase treatment of albumin, S-[2 [(hydroxyethyl)thio]ethyl]Cys-Pro-Phe was analyzed by means of micro-LC/tandem MS, allowing a detection limit for in vitro exposure of human blood of 10 nM, which is 1 order of magnitude lower than that obtained by means of modified Edman degradation. The analytical procedure could be successfully applied to the analysis of albumin samples from Iranian victims of the Iran-Iraq war. PMID- 10458706 TI - A gas chromatography/electron capture/negative chemical ionization high resolution mass spectrometry method for analysis of endogenous and exogenous N7 (2-hydroxyethyl)guanine in rodents and its potential for human biological monitoring. AB - A gas chromatography/electron capture/negative chemical ionization high resolution mass spectrometry (GC/EC/NCI-HRMS) method was developed for quantitating N7-(2-hydroxyethyl)guanine (N7-HEG) with excellent sensitivity and specificity. [4,5,6,8-(13)C(4)]-N7-HEG was synthesized, characterized, and quantitated using HPLC/electrospray ionization mass spectrometry (HPLC/ESI-MS) so it could serve as an internal standard. After being converted to its corresponding xanthine and derivatized with pentafluorobenzyl (PFB) bromide twice, the PFB derivative of N7-HEG was characterized using GC/EC/NCI-HRMS carried out at full scan mode. The most abundant fragment was at m/z 555, with a molecular formula of C(21)H(9)N(4)O(3)F(10), resulting from the loss of one PFB group. By monitoring m/z 555.0515 (analyte) and m/z 559.0649 (internal standard), this assay demonstrated a linear relationship over a range of 1 fmol to 1 pmol of N7-HEG versus 20 fmol of [(13)C(4)]-N7-HEG on column. The limit of detection (LOD) for the complete assay was 600 amol (S/N = 5) injected on column. The variation of this assay was within 15% from 1 to 20 fmol of N7-HEG versus 2 fmol of [(13)C(4)]-N7-HEG with four replications for each calibration standard. Two hundred to three hundred micrograms of spleen DNA of control rats and mice and 100 microg of spleen DNA of rats and mice exposed to 3000 ppm ethylene for 6 h/day for 5 days were analyzed using GC/EC/NCI-HRMS. The amounts of N7-HEG varied from 0.2 to 0.3 pmol/micromol of guanine in tissues of control rats. Ethylene exposed animals had 5-15-fold higher N7-HEG levels than controls. This assay was able to quantitate N7-HEG in 25-30 microg of DNA from human lymphocytes with excellent specificity. This was due in part to human tissues having 10-15-fold higher amounts of endogenous N7-HEG than rodents. These results show that this GC/EC/NCI-HRMS method is highly sensitive and specific and can be used in biological monitoring and molecular dosimetry and molecular epidemiology studies. PMID- 10458707 TI - Metabolism of carbon tetrachloride to trichloromethyl radical: An ESR and HPLC-EC study. AB - Extensive ESR spin-trapping studies with alpha-phenyl-N-tert-butylnitrone (PBN) have shown that carbon tetrachloride (CCl(4)) is metabolized to trichloromethyl radical ((*)CCl(3)). However, the ESR analysis of alpha-phenyl-N-tert butylnitrone (PBN)-spin trapped (*)CCl(3) in biological systems appears to be complicated. It has been reported that after in vivo administration of PBN and CCl(4) to rats, most of the PBN-CCl(3) adduct collected in the bile was ESR silent, suggesting reduction of the nitroxide to its hydroxylamine form. The PBN CCl(3) nitroxide was also shown to undergo a NADPH-dependent reduction in the presence of liver microsomes. Thus, it appears that the variability (or the absence) of the ESR signal of PBN-CCl(3) nitroxide in biological systems reflects, at least in part, the fluctuations in the equilibrium between the nitroxide and hydroxylamine forms of this adduct. To test this possibility, ESR and HPLC experiments with electrochemical detection (EC) were conducted for analysis of the major redox form of the PBN-CCl(3) adduct in vivo. Standard procedures for the in vitro preparation of both redox forms of PBN-CCl(3) and for their HPLC-EC analysis and electrochemical profiles were established. The intensity of the initially observed ESR spectrum of PBN-CCl(3) nitroxide of the liver extract from a CCl(4)- and PBN-treated rat was relatively constant; after an addition of K(3)[Fe(CN)(6)] to the extract, the intensity of the ESR spectrum increased by 1 order of magnitude, most likely due to the co-oxidation of ESR silent PBN-derived hydroxylamines. The addition of PBN-CCl(3) nitroxide to the liver homogenate resulted in the rapid loss of the ESR signal. The HPLC-EC analysis of the liver extract revealed that the in vivo spin trapping of (*)CCl(3) with PBN leads to a preferential formation of the ESR silent PBN-CCl(3) hydroxylamine. The predominant presence of the hydroxylamine derivative was also detected in the blood of a CCl(4)-treated rat. The results of this work are discussed in terms of combination of the ESR spin trapping and HPLC-EC techniques for the detection of ESR silent radical adducts in biological systems. PMID- 10458708 TI - Relationship between surface properties and cellular responses to crystalline silica: studies with heat-treated cristobalite. AB - A fibrogenic sample of cristobalite dust, CRIS (crystalline silica of mineral origin), was heated to 1300 degrees C (CRIS-1300) to relate induced physicochemical modifications to cytotoxicity. Heating did not affect dust micromorphology and crystallinity, except for limited sintering and decreased surface area of CRIS-1300. Thermal treatments deeply affected surface properties. Electron paramagnetic resonance showed surface radicals progressively annealed by heating, mostly disappearing at >/=800 degrees C. Surface hydrophilicity or hydrophobicity, evaluated with water vapor adsorption, still showed some hydrophilic patches in CRIS-800, but CRIS-1300 was fully hydrophobic. Heating modified the biological activity of cristobalite. Cytotoxicity, tested on proliferating cells of the mouse monocyte macrophage cell line J774, showed that CRIS was cytotoxic and CRIS-800 was still cytotoxic, but CRIS-1300 was substantially inert. Cytotoxicity of CRIS to the rat lung alveolar epithelial cell line, AE6, as measured by colony forming efficiency, was greatly reduced for CRIS-800 and eliminated for CRIS-1300. The rate of lactate dehydrogenase release by rat alveolar macrophages was lowered for CRIS-800, and release was completely inactivated for CRIS-1300. The absence of surface radicals and the onset of hydrophobicity may both account for the loss of cytotoxicity upon heating. Differences observed between CRIS-800 and CRIS-1300, both fully deprived of surface radicals, indicate that hydrophobicity is at least one of the surface properties determining the cytotoxic potential of a dust. PMID- 10458709 TI - Late thrombosis after radiation. Sitting on a time bomb. PMID- 10458710 TI - Angiogenesis and atherosclerosis. The mandate broadens. PMID- 10458711 TI - Cardiac pacing. An alternative treatment for selected patients with hypertrophic cardiomyopathy and adjunctive therapy for certain patients with dilated cardiomyopathy. PMID- 10458712 TI - Late coronary occlusion after intracoronary brachytherapy. AB - BACKGROUND: Intracoronary brachytherapy appears to be a promising technology to prevent restenosis. Presently, limited data are available regarding the late safety of this therapeutic modality. The aim of the study was to determine the incidence of late (>1 month) thrombosis after PTCA and radiotherapy. METHODS AND RESULTS: From April 1997 to March 1999, we successfully treated 108 patients with PTCA followed by intracoronary beta-radiation. Ninety-one patients have completed at least 2 months of clinical follow-up. Of these patients, 6.6% (6 patients) presented with sudden thrombotic events confirmed by angiography 2 to 15 months after intervention (2 balloon angioplasty and 4 stent). Some factors (overlapping stents, unhealed dissection) may have triggered the thrombosis process, but the timing of the event is extremely unusual. Therefore, the effect of radiation on delaying the healing process and maintaining a thrombogenic coronary surface is proposed as the most plausible mechanism to explain such late events. CONCLUSIONS: Late and sudden thrombosis after PTCA followed by intracoronary radiotherapy is a new phenomenon in interventional cardiology. PMID- 10458713 TI - Increased proinflammatory cytokines in patients with chronic stable angina and their reduction by aspirin. AB - BACKGROUND: Proinflammatory cytokines released by injured endothelium facilitate interaction of endothelial cells with circulating leukocytes and thus may contribute to development and progression of atherosclerosis. We investigated whether cytokines and C-reactive protein (CRP) are indicative of myocardial ischemia or of diseased vessels and whether they are influenced by aspirin treatment in patients with chronic stable angina. METHODS AND RESULTS: Plasma macrophage colony stimulating factor (MCSF), IL-1b, IL-6, and CRP were measured in 60 stable patients after 48-hour Holter monitoring and in 24 matched controls. All patients had angiographic documentation of disease and positive exercise ECGs. Patients with ischemia on Holter monitoring (n=40) received aspirin or placebo in a 6-week, randomized, double blind, crossover trial. Blood sampling was repeated at the end of each treatment phase (3 weeks). Compared to controls, patients had more than twice median MCSF (800 versus 372 pg/mL), IL-6 (3.9 versus 1.7 pg/mL), and CRP (1.25 versus 0.23 mg/L) levels (P<0.01 for all comparisons). MCSF was related to ischemia on Holter monitoring (P<0.01), to low ischemic threshold during exercise (P<0.01), and together with IL-1b to number of diseased vessels (P<0.05). MCSF, IL-6, and CRP were all reduced after 6 weeks of aspirin treatment (P<0.05). CONCLUSIONS: These findings suggest that cytokines are associated with both ischemia and anatomic extent of disease in patients with stable angina. Reduced cytokine and CRP levels by aspirin may explain part of aspirin's therapeutic action. PMID- 10458714 TI - Acute platelet inhibition with abciximab does not reduce in-stent restenosis (ERASER study). The ERASER Investigators. AB - BACKGROUND: Although stents reduce restenosis compared with balloon angioplasty, their long-term efficacy is limited by neointimal hyperplasia. Platelet and alpha(v)beta(3) integrin receptor inhibition limits neointimal proliferation in animal models of arterial injury. METHODS AND RESULTS: We tested whether the dual beta(3) integrin blocking agent abciximab, administered for 12 or 24 hours at the same intravenous dose as that shown to reduce adverse clinical events (death, infarction, and revascularization) after angioplasty, would reduce restenotic tissue volume, as measured by intravascular ultrasound at 6 months. Two hundred twenty-five patients were randomly allocated to placebo or abciximab before coronary intervention. Of the 215 patients who received stents and study drug, 191 (88.8%) returned for late (>/=4 months) coronary evaluation. Tissue volume, expressed as a percentage of stent volume, did not differ: 25+/-15%, 27+/-15%, and 29+/-14% for the patients in the placebo and the 12- and 24-hour abciximab groups, respectively. Lack of abciximab benefit was confirmed by quantitative coronary angiography (dichotomous restenosis: 11.6%, 18. 9%, and 19.4%; loss index: 0.33, 0.52, and 0.47, respectively, P=NS). CONCLUSIONS: Potent platelet inhibition with abciximab, as administered in this study, does not reduce in stent restenosis. The interrelationship between stents, platelets, and neointimal proliferation requires further study. PMID- 10458715 TI - Ventricular pacing with premature excitation for treatment of hypertensive cardiac hypertrophy with cavity-obliteration. AB - BACKGROUND: Hypertensive left ventricular hypertrophy with supranormal systolic ejection and distal cavity obliteration (HHCO) can result in debilitating exertional fatigue and dyspnea. Dual-chamber pacing with ventricular preactivation generates discoordinate contraction, which can limit cavity obliteration and thereby increase potential ejection reserve. Accordingly, we hypothesized that pacing may improve exercise tolerance long-term in this syndrome. METHODS AND RESULTS: Dual-chamber pacemakers were implanted in 9 patients with exertional dyspnea caused by HHCO. Intrinsic atrial rate was sensed, and ventricular preactivation was achieved by shortening the atrial ventricular delay. Pacing was on or off for successive 3-month periods (randomized, double-blind, crossover design), followed by 6 additional pacing-on months. Metabolic exercise testing, quality-of-life assessment, and rest and dobutamine-stress echocardiographic/Doppler data were obtained. After 3 months of pacing-on, exercise duration rose from 324+/-133 to 588+/-238 s (mean+/-SD; P=0.001, with 7 of 9 patients improving >/=30%), and maximal oxygen consumption increased from 13.6+/-2.9 to 16.7+/-3.3 mL of O(2). min(-1). kg(-1) (P<0.02). Both parameters were little changed from baseline during the pacing-off period. Improved exercise capacity persisted at 1-year follow-up. Clinical symptoms and activities of daily living improved during the pacing-on period and stayed improved at 1 year, but they were little changed during the pacing-off period. Despite similar basal values, stroke volume (P<0.001) and cardiac output (P<0.02) increased with dobutamine stimulation 2 to 3 times more after 1 year of follow-up as compared with baseline. CONCLUSIONS: Long-term dual-chamber pacing can improve exercise capacity, cardiac reserve, clinical symptoms, and activities of daily living in patients with HHCO. This therapy may provide a novel alternative for patients in whom traditional pharmacological treatment proves inadequate. PMID- 10458716 TI - Effects of autonomic neuropathy on coronary blood flow in patients with diabetes mellitus. AB - BACKGROUND: C ardiac sympathetic signals play an important role in the regulation of myocardial perfusion. We hypothesized that sympathetically mediated myocardial blood flow would be impaired in diabetics with autonomic neuropathy. METHODS AND RESULTS: We studied 28 diabetics (43+/-7 years old) and 11 age-matched healthy volunteers. PET was used to delineate cardiac sympathetic innervation with [(11)C]hydroxyephedrine ([(11)C]HED) and to measure myocardial blood flow at rest, during hyperemia, and in response to sympathetic stimulation by cold pressor testing. The response to cardiac autonomic reflex tests was also evaluated. Using ultrasonography, we also measured brachial artery reactivity during reactive hyperemia (endothelium-dependent dilation) and after sublingual nitroglycerin (endothelium-independent dilation). Based on [(11)C]HED PET, 13 of 28 diabetics had sympathetic-nerve dysfunction (SND). Basal flow was regionally homogeneous and similar in the diabetic and normal subjects. During hyperemia, the increase in flow was greater in the normal subjects (284+/-88%) than in the diabetics with SND (187+/-80%, P=0.084) and without SND (177+/-72%, P=0.028). However, the increase in flow in response to cold was lower in the diabetics with SND (14+/-10%) than in those without SND (31+/-12%) (P=0.015) and the normal subjects (48+/-24%) (P<0.001). The flow response to cold was related to the myocardial uptake of [(11)C]HED (P<0.001). Flow-mediated brachial artery dilation was impaired in the diabetics compared with the normal subjects, but it was similar in the diabetics with and without SND. CONCLUSIONS: Diabetic autonomic neuropathy is associated with an impaired vasodilator response of coronary resistance vessels to increased sympathetic stimulation, which is related to the degree of SND. PMID- 10458718 TI - Fully discharging phases. A new approach to biphasic waveforms for external defibrillation. AB - BACKGROUND: Phase-2 voltage and maximum pulse width are dependent on phase-1 pulse characteristics in a single-capacitor biphasic waveform. The use of 2 separate output capacitors avoids these limitations and may allow waveforms with lower defibrillation thresholds. A previous report also suggested that the optimal tilt may be >70%. This study was designed to determine an optimal biphasic waveform by use of a combination of 2 separate and fully (95% tilt) discharging capacitors. METHODS AND RESULTS: We performed 2 external defibrillation studies in a pig ventricular fibrillation model. In group 1, 9 waveforms from a combination of 3 phase-1 capacitor values (30, 60, and 120 microF) and 3 phase-2 capacitor values (0=monophasic, 1/3, and 1.0 times the phase-1 capacitor) were tested. Biphasic waveforms with phase-2 capacitors of 1/3 times that of phase 1 provided the highest defibrillation efficacy (stored energy and voltage) compared with corresponding monophasic and biphasic waveforms with the same capacitors in both phases except for waveforms with a 30-microF phase-1 capacitor. In group 2, 10 biphasic waveforms from a combination of 2 phase-1 capacitor values (30 and 60 microF) and 5 phase-2 capacitor values (10, 20, 30, 40, and 50 microF) were tested. In this range, phase-2 capacitor size was more critical for the 30-microF phase-1 than for the 60-microF phase-1 capacitor. The optimal combinations of fully discharging capacitors for defibrillation were 60/20 and 60/30 microF. Conclusions-Phase-2 capacitor size plays an important role in reducing defibrillation energy in biphasic waveforms when 2 separate and fully discharging capacitors are used. PMID- 10458717 TI - Insulin stimulates both endothelin and nitric oxide activity in the human forearm. AB - BACKGROUND: The mechanism of the hemodynamic effect of insulin in the skeletal muscle circulation has not been fully elucidated. The purpose of this study was to assess whether the hemodynamic response to insulin involves the concurrent release of endothelin (ET-1) and nitric oxide (NO), 2 substances with opposing vasoactive properties. METHODS AND RESULTS: Bioactivity of ET-1 and NO was assessed without insulin and during insulin infusion in the forearm circulation of healthy subjects by use of blockers of ET-1 receptors and by NO synthesis inhibition. In the absence of hyperinsulinemia, ET-1 receptor blockade did not result in any significant change in forearm blood flow from baseline (P=0.29). Intra-arterial insulin administration did not significantly modify forearm blood flow (P=0. 88). However, in the presence of hyperinsulinemia, ET-1 receptor antagonism was associated with a significant vasodilator response (P<0.001). In the presence of ET-1 receptor blockade, the vasoconstrictor response to NO inhibition by N(G)-monomethyl-L-arginine was significantly higher after insulin infusion than in the absence of hyperinsulinemia (P=0.006). CONCLUSIONS: These findings suggest that in the skeletal muscle circulation, insulin stimulates both ET-1 and NO activity. An imbalance between the release of these 2 substances may be involved in the pathophysiology of hypertension and atherosclerosis in insulin resistant states associated with endothelial dysfunction. PMID- 10458719 TI - Chlamydia pneumoniae infection and early asymptomatic carotid atherosclerosis. AB - BACKGROUND: Chronic Chlamydia pneumoniae infection has been implicated in the pathogenesis of atherosclerosis but whether it plays a role at an early stage in the disease is uncertain. An early estimate of atherosclerosis can be obtained by ultrasonic imaging of the carotid artery to determine intima-media thickness (IMT) and the thickness of any atheroma plaques. METHODS AND RESULTS: In 983 normal population individuals aged 30 to 70 years, we measured common carotid artery (CCA) and carotid bulb IMT, and also carotid plaque thickness and the degree of internal carotid artery (ICA) stenosis. C. pneumoniae IgA titers of >/=16 and IgG titers of >/=64 were taken as positive. There was no association between C. pneumoniae IgA or IgG seropositivity with right, left, or mean CCA or bulb IMT, or with the presence of carotid plaques. There was a significant association between IgA seropositivity and >50% mean carotid stenosis with an odds ratio of 5.24 (95% CI 1.24 to 22.21, P=0.0245) after controlling for age and sex; after controlling for other cardiovascular risk factors, this was not significant 3.96 (95% CI 0. 84 to 18.78, P=0.082). No association was found between IgA or IgG seropositivity and markers of fibrinogen, log C-reactive protein, or leukocyte count. CONCLUSIONS: We found no evidence that serological evidence of C. pneumoniae infection is associated with early atherosclerosis. It is possible that IgA seropositivity is associated with more advanced disease but this hypothesis needs to be examined in a population with a higher prevalence of advanced atherosclerosis. We found no evidence that C. pneumoniae results in a chronic systemic inflammatory state. PMID- 10458720 TI - Increased carotid intimal-medial thickness and coronary calcification are related in young and middle-aged adults. The Muscatine study. AB - BACKGROUND: Increased carotid intimal-medial thickness (IMT) and coronary artery calcification (CAC) are used as 2 markers of early atherosclerosis. Our objectives were to assess whether increased IMT and CAC are related and to determine the relationship between cardiovascular risk factors and carotid IMT in young adults. METHODS AND RESULTS: A sample of 182 men and 136 women aged 33 to 42 years living in Muscatine, Iowa, underwent B-mode carotid ultrasound to determine the mean of 12 measurements of maximal carotid IMT. CAC was defined as calcification in the proximal coronary arteries in >/=3 contiguous pixels with a density of >/=130 HU. The mean IMT was 0.788 mm (SD 0.127) for men and 0.720 mm (SD 0.105) for women. CAC was present in 27% of men and 14% of women and was significantly associated with IMT in men (P<0.025) and women (P<0.005). With multivariate analysis, after adjustment for age, significant risk factors for carotid IMT were LDL cholesterol (P<0.001) and pack-years of smoking (P<0.05) in men and LDL cholesterol (P<0.001) and systolic blood pressure (P<0.01) in women. These risk factors remained significant after CAC was included in the multivariate model. CONCLUSIONS: There is an association between increased carotid IMT and CAC and between cardiovascular risk factors and increased IMT in young adults. Carotid IMT may provide information in addition to CAC that can be used to identify young adults with premature atherosclerosis. PMID- 10458721 TI - Brief myocardial ischemia attenuates platelet thrombosis in remote, damaged, and stenotic carotid arteries. AB - BACKGROUND: Brief antecedent periods of coronary artery occlusion improve subsequent vessel patency in damaged and stenotic coronary arteries via release of adenosine from ischemic/reperfused myocardium and resultant adenosine receptor stimulation. However, the site of receptor stimulation-circulating blood-borne elements (ie, platelets) versus vessel-wall components of the culprit artery remains unclear. If platelet adenosine receptors are involved, then the benefits of brief coronary occlusion (1) should be manifested systemically and improve patency at a remote site and (2) should be inhibited by an antagonist of adenosine A(2) receptors, whereas, in contrast, (3) brief vascular occlusion not associated with appreciable adenosine release should be ineffective in improving vessel patency. METHODS AND RESULTS: In Protocol 1, anesthetized rabbits received 5 minutes of transient coronary occlusion, 5 minutes of transient bilateral carotid occlusion (purported to cause negligible adenosine release from the brain), or no intervention. All rabbits then underwent injury plus stenosis of the left carotid artery, resulting in repeated cyclic variations in carotid blood flow (CFVs). Carotid patency during the initial 2 hours after stenosis (assessed by quantifying the nadir of the CFVs and area of the flow-time profile) was significantly enhanced with antecedent coronary-but not carotid-occlusion versus controls. In Protocol 2, improvement in carotid patency after brief coronary occlusion was corroborated in anesthetized dogs. However, the benefits of brief coronary occlusion were abrogated by the A(2)/A(1) antagonist CGS 15943. CONCLUSIONS: Brief antecedent coronary artery occlusion enhanced vessel patency in remote, damaged, and stenotic carotid arteries, largely due to adenosine receptor stimulation on circulating elements. PMID- 10458722 TI - Antibody blockade of thrombospondin accelerates reendothelialization and reduces neointima formation in balloon-injured rat carotid artery. AB - BACKGROUND: Remodeling of the extracellular matrix plays an important role during the pathogenesis of atherosclerosis and restenosis. The matrix glycoprotein thrombospondin-1 (TSP1) inhibits endothelial cell proliferation and migration in vitro. In contrast, TSP1 facilitates the growth and migration of cultured vascular smooth muscle cells. Accordingly, we investigated the hypothesis that administration of anti-TSP1 antibody could facilitate reendothelialization and inhibit neointimal thickening in balloon-injured rat carotid artery. METHODS AND RESULTS: Sprague-Dawley rats were subjected to left common carotid artery denudation, after which arteries were treated with C6.7 anti-TSP1 or control antibody. Evans blue dye staining 2 weeks after injury disclosed significantly increased reendothelialization in arteries treated with C6.7 antibody compared with the control group, and this effect was associated with increased number of proliferating cell nuclear antigen-positive endothelial cells. In contrast, treatment with C6.7 antibody decreased the number of proliferating cell nuclear antigen-positive vascular smooth muscle cells in the injured arterial wall. Neointimal thickening was correspondingly attenuated to a statistically significant degree in arteries receiving C6.7 antibody versus the control group at both the 2-week and 4-week time points. CONCLUSIONS: Intra-arterial delivery of antibody against TSP1 facilitated reendothelialization and reduced neointimal lesion formation after balloon denudation. PMID- 10458723 TI - Aggregating human platelets stimulate expression of vascular endothelial growth factor in cultured vascular smooth muscle cells through a synergistic effect of transforming growth factor-beta(1) and platelet-derived growth factor(AB). AB - BACKGROUND: Vascular endothelial growth factor (VEGF), an endothelial mitogen and chemoattractant, has been implicated in the recovery of the endothelium after balloon injury. The increased expression of VEGF in vascular smooth muscle cells (SMC) at sites of injury suggests that this cell type may be a major cellular source of VEGF. This study examined whether aggregating platelets stimulate VEGF expression in cultured SMC. METHODS AND RESULTS: ++VEGF expression in SMC was assessed by Northern blot analysis and by reverse transcription followed by polymerase chain reaction and the release of VEGF by Western blot analysis and immunoassay. Platelet-derived products (PDP) released by aggregating human platelets time-dependently and concentration-dependently enhanced VEGF mRNA levels, mainly that coding for the soluble splice variant VEGF(165/164), and stimulated the release of VEGF protein. These effects were potentiated by transient acidification of PDP, which release bioactive transforming growth factor (TGF)-beta(1), and mimicked by platelet-derived growth factor (PDGF)(AB) and TGF-beta(1) in a synergistic manner. Both a TGF-beta-neutralizing antibody and a PDGF-neutralizing antibody significantly attenuated the effect of acidified PDP on VEGF production. CONCLUSIONS: Aggregating human platelets induce VEGF mRNA expression in cultured SMC and the subsequent release of VEGF protein. This effect can be attributed to a supra-additive action of PDGF(AB) and TGF-beta(1) and may represent a novel mechanism by which platelets contribute to the recovery of the endothelial lining at sites of balloon-injured arteries. PMID- 10458724 TI - Inhibition of neointima hyperplasia of mouse vein grafts by locally applied suramin. AB - BACKGROUND: Saphenous vein grafts are widely used for aortocoronary bypass surgery as treatment for severe atherosclerosis and often are complicated by subsequent occlusion of the graft vessel. METHODS AND RESULTS: We described a mouse model of venous bypass graft arteriosclerosis that can be effectively retarded by locally applied suramin, a growth factor receptor antagonist. Mouse isogeneic vessels of the vena cava veins pretreated with suramin were grafted end to end into the carotid arteries and enveloped with a mixture of suramin (1 mmol/L) and pluronic-127 gel. In the untreated group, vessel wall thickening was observed as early as 1 week after surgery and progressed to 4-fold and 10-fold the original thickness in grafted veins at 4 and 8 weeks, respectively. Pluronic 127 gel alone did not influence neointima formation. Suramin treatment reduced the neointima hyperplasia 50% to 70% compared with untreated controls. Immunohistochemical studies demonstrated that a significant proliferation of vascular smooth muscle cells (SMCs) constituted neointimal lesions between 4 and 8 weeks. The majority of SMCs expressed platelet-derived growth factor (PDGF) receptors-alpha and -beta, which were significantly reduced by suramin treatment. In vitro studies indicated that suramin completely blocked PDGF receptor activation or phosphorylation stimulated by PDGF-AB, inhibited activation of mitogen-activated protein kinase (ERK) kinases (MEK1/2) and ERK1/2, and abrogated transcription factor AP-1 DNA-binding activity. CONCLUSIONS: Suramin inhibited SMC migration and proliferation in vivo and in vitro by blocking PDGF-initiated PDGF receptor and MAPK-AP-1 signaling. These findings indicate that locally applied suramin is effective in a mouse model of venous bypass graft arteriosclerosis. PMID- 10458725 TI - Aminorex, fenfluramine, and chlorphentermine are serotonin transporter substrates. Implications for primary pulmonary hypertension. AB - BACKGROUND: Coadministration of phentermine and fenfluramine (phen/fen) effectively treats obesity and possibly addictive disorders. The association of fenfluramine and certain other anorexic agents with serious side effects, such as cardiac valvulopathy and primary pulmonary hypertension (PPH), limits the clinical utility of these drugs. Development of new medications that produce neurochemical effects like phen/fen without causing unwanted side effects would be a significant therapeutic breakthrough. METHODS AND RESULTS: We tested the hypothesis that fenfluramine (and other anorexic agents) might increase the risk of PPH through interactions with serotonin (5-HT) transporters. Because 5-HT transporter proteins in the lung and brain are identical, we examined, in rat brain, the effects of selected drugs on 5-HT efflux in vivo and monoamine transporters in vitro as a generalized index of transporter function. Our data show that drugs known or suspected to increase the risk of PPH (eg, aminorex, fenfluramine, and chlorphentermine) are 5-HT transporter substrates, whereas drugs that have not been shown to increase the risk of PPH are less potent in this regard. CONCLUSIONS: We speculate that medications that are 5-HT transporter substrates get translocated into pulmonary cells where, depending on the degree of drug retention, their intrinsic drug toxicity, and individual susceptibility, PPH could develop as a response to high levels of these drugs or metabolites. Emerging evidence suggests that it is possible to develop transporter substrates devoid of adverse side effects. Such medications could have therapeutic application in the management of obesity, drug dependence, depression, and other disorders. PMID- 10458726 TI - Relation between ligament of Marshall and adrenergic atrial tachyarrhythmia. AB - BACKGROUND: The mechanism of the adrenergic atrial tachyarrhythmia is unclear. We hypothesize that the ligament of Marshall (LOM) is sensitive to adrenergic stimulation and may serve as a source of the adrenergic atrial tachyarrhythmia. METHODS AND RESULTS: We performed computerized mapping studies in isolated perfused canine left atrial tissues from normal dogs (n=9) and from dogs with chronic atrial fibrillation (AF) induced by 10 to 41 weeks of rapid pacing (n=3). Before isoproterenol, spontaneous activity occurred in only one normal tissue (cycle length, CL >1300 ms). During isoproterenol infusion, automatic rhythm was induced in both normal tissues (CL=578+/-172 ms) and AF tissues (CL=255+/-29 ms, P<0.05). The origin of spontaneous activity was mapped to the LOM. In the AF tissues, but not the normal tissues, we observed the transition from rapid automatic activity to multiple wavelet AF. Ablation of the LOM terminated the spontaneous activity and prevented AF. Immunocytochemical studies of the LOM revealed muscle tracts surrounded by tyrosine hydroxylase-positive (sympathetic) nerves. CONCLUSIONS: We conclude that the LOM is richly innervated by sympathetic nerves and serves as a source of isoproterenol-sensitive focal automatic activity in normal canine atrium. The sensitivity to isoproterenol is upregulated after long-term rapid pacing and may contribute to the development of AF in this model. PMID- 10458727 TI - Images in cardiovascular medicine. 21st-century imaging for a 19th-century disease. PMID- 10458728 TI - ACC/AHA guidelines for ambulatory electrocardiography: executive summary and recommendations. A report of the American College of Cardiology/American Heart Association task force on practice guidelines (committee to revise the guidelines for ambulatory electrocardiography). PMID- 10458729 TI - "Reel Syndrome": a new form of Twiddler's syndrome? PMID- 10458731 TI - Laparoscopic fundoplication for dysphagia and peptic esophageal stricture. AB - Peptic esophageal stricture with dysphagia is a late manifestation of severe gastroesophageal reflux disease (GERD). Although laparoscopic fundoplication is an effective antireflux operation, its efficacy for persons with peptic esophageal stricture and dysphagia has not been well defined. The aim of this study was to evaluate outcomes after fundoplication in this subgroup of GERD patients. Forty GERD patients with moderate, severe, or incapacitating dysphagia and peptic esophageal stricture were compared to a control group of 121 GERD patients without significant dysphagia or stricture. Reflux symptom severity was scored by each patient preoperatively and at most recent follow-up postoperatively (mean 1.5 years) using a scale ranging from 0 to 4 (0 = symptoms absent; 4 = symptoms incapacitating). Symptom scores were compared by the Wilcoxon rank-sum test. Postoperative redilation and fundoplication failure rates were also determined. At a mean follow-up of 1.5 years after fundoplication, the median dysphagia score had improved from 3 to 0 (P <0.001) in stricture patients and remained low (score 0) in the control group. The median heartburn score also improved from 3 to 0 (P <0.001) in stricture patients, with an identical response in the control group (P <0.001). Among dysphagia/stricture patients, 35 (87.5%) reported overall satisfaction and have not required secondary medical treatment or esophageal dilation. Four patients (10%) have required endoscopic redilation for residual dysphagia and one (2.5%) had reoperation for fundoplication herniation shortly after operation. Laparoscopic fundoplication is an effective therapy for patients with dysphagia and peptic esophageal stricture. PMID- 10458730 TI - Intestinal microcirculation and gut permeability in acute pancreatitis: early changes and therapeutic implications. AB - Translocation of bacteria from the intestine causes local and systemic infection in severe acute pancreatitis. Increased intestinal permeability is considered a promoter of bacterial translocation. The mechanism leading to increased gut permeability may involve impaired intestinal capillary blood flow. The aim of this study was to evaluate and correlate early changes in capillary blood flow and permeability of the colon in acute rodent pancreatitis of graded severity. Edematous pancreatitis was induced by intravenous cerulein; necrotizing pancreatitis by intravenous cerulein and intraductal glycodeoxycholic acid. Six hours after induction of pancreatitis, the permeability of the ascending colon was assessed by the Ussing chamber technique; capillary perfusion of the pancreas and colon (mucosal and subserosal) was determined by intravital microscopy. In mild pancreatitis, pancreatic capillary perfusion remained unchanged (2.13 c 0.06 vs. 1.98 +/-0.04 nl x min(-1) x cap(-1) [control]; P = NS), whereas mucosal (1.59 +/-0.03 vs. 2.28 +/-0.03 nl x min(-1) x cap((-1))[control]; P <0.01) and subserosal (2.47 +/-0.04 vs. 3.74 +/-0.05 nl x min(-1) x cap((-1))[control]; P <0.01) colonic capillary blood flow was significantly reduced. Severe pancreatitis was associated with a marked reduction in both pancreatic (1.06 +/ 0.03 vs. 1.98 +/-0.04 nl x min(-1) x cap(-1) [control]; P <0. 01) and colonic (mucosal: 0.59 +/-0.01 vs. 2.28 +/-0.03 nl x min(-1) x cap((-1))[control], P <0.01; subserosal: 1.96 +/-0.05 vs. 3.74 +/-0.05 nl x min(-1) x cap(-1) [control], P <0.01) capillary perfusion. Colon permeability tended to increase with the severity of the disease (control: 147 +/-19 nmol x thr(-1) x cm(-2); mild pancreatitis: 158 +/-23 nmol x hr(-1) x cm(-2); severe pancreatitis: 181 +/ 33 nmol x hr(-1) x cm(-2); P = NS). Impairment of colonic capillary perfusion correlates with the severity of pancreatitis. A decrease in capillary blood flow in the colon, even in mild pancreatitis not associated with significant protease activation and acinar cell necrosis or impairment of pancreatic capillary perfusion, suggests that colonic microcirculation is especially susceptible to inflammatory injury. There was no significant change in intestinal permeability in the early stage of pancreatitis, suggesting a window of opportunity for therapeutic interventions to prevent the later-observed increase in gut permeability, which could result in improved intestinal microcirculation. PMID- 10458732 TI - Analysis of cell growth kinetics and substrate diffusion in a polymer scaffold. AB - The cultivation of cartilage cells (chondrocytes) in polymer scaffolds leads to implants that may potentially be used to repair damaged joint cartilage or for reconstructive surgery. For this technique to be medically applicable, the physical parameters that govern cell growth in a polymer scaffold must be understood. This understanding of cell behavior under in vitro conditions, where diffusion is the primary mode of transport of nutrients, may aid in the scale-up of the cartilage generation process. A mathematical model of chondrocyte generation and nutrient consumption is developed here to analyze the behavior of cell growth in a biodegradable polymer matrix for a series of different thickness polymers. Recent literature has implied that the diffusion of nutrients is a major factor that limits cell growth (Freed et al., 1994). In the present paper, a mathematical model is developed to directly relate the effects of increasing cell mass in the polymer matrix on the transport of nutrients. Reaction and diffusion of nutrients in the cell-polymer system are described using the fundamental species continuity equations and the volume averaging method. The volume averaging method is utilized to derive a single averaged nutrient continuity equation that includes the effective transport properties. This approach allows for the derivation of effective diffusion and rate coefficients as functions of the cell volume fraction. The cell volume fraction as a function of time is determined by solution of a material balance on cell mass. Growth functions including the Moser, a modified Contois, and an nth-order heterogeneous growth kinetic model are evaluated through a parameter analysis, and the results are compared to experimental data found in the literature. The results indicate that cellular functions in conjunction with mass transfer processes can account partially for the general trends in the cell growth behavior for various thickness polymers. The Contois growth function appeared to describe the data more accurately in terms of the lag period at early times and the long time limits. However, all kinetic growth functions required variations in the kinetic parameters to fully describe the effects of polymer thickness. This result implies that restricted diffusion of nutrients is not the sole factor limiting cell growth when the thickness of the polymer is changed. Therefore, further experimental data and model improvements are needed to accurately describe the cell growth process. PMID- 10458733 TI - Proteolytic activity in infected and noninfected insect cells: degradation of HIV 1 Pr55gag particles. AB - In this work the proteolytic activity in the supernatant and inside insect cells in culture was evaluated for different multiplicities of infection (MOI) and times of infection (TOI). Several methods to detect proteolytic activity in insect cells were tested and that using fluorescein thiocyanite-casein as a substrate was chosen. It was observed that infection caused not only a reduction in the concentration of proteases by decreasing their synthesis but also an inhibition of the intracellular proteolytic activity by increasing the intracellular ATP level (measured by in vivo nuclear magnetic resonance, NMR). The maximum proteolytic activity in the supernatant was observed at 72 hpi except when the cells were infected in the late exponential growth phase or with very low MOI, yielding a nonsynchronous infection. The proteolytic degradation of Pr55gag particles was studied during culture and after harvest. In this particular case it was concluded that the supernatant should be stored at low temperature or quickly purified, since the degradation after 24 h is only 3% at 4 degrees C while at 27 degrees C this value rises to 23%. There is a complex relationship between MOI, TOI, proteolytic activity, and product titer and quality. Thus, the optimal conditions for each case will be a compromise between the final product titer, the desired product quality, and operational issues like process time and capacity, requiring proper integration between bioreaction and downstream processing. PMID- 10458734 TI - A novel autoregulated proliferation-controlled production process using recombinant CHO cells. AB - Controlled proliferation bioprocesses have shown great enhancement of heterologous protein production. This novel technology has been implemented here using a multicistronic expression unit encoding the product gene and a cytostatic cell-cycle-arresting gene (p27) under control of a single tetracycline repressible (tet(off)) promoter. The strict genetic linkage of both genes allows the dissection of the production process into a nonproductive growth phase (dicistronic expression unit repressed) followed by a proliferation-inhibited production phase (dicistronic expression unit induced) when the cells have reached an optimal cell density. Based on rapid degradation of the external repressible agents tetracycline (tet) and doxycycline (dox) in the cell culture medium, we developed a self-regulated process for transition from the growth phase to the production phase in a fashion that is dependent only on the starting cell population and the initial concentration of the tetracyclines. With this process, no change in medium is required to accomplish the transition from growth to production phase. The two-phase bioprocess achieved here by tet switch controlled proliferation is reliable and allows a growth-arrested production phase of at least 7 days, during which cells remain in a well-defined, highly viable physiological state and show enhanced heterologous protein production. This Tet(SWITCH) process is readily adaptable to a variety of industrial processes designed for production of difficult-to-express protein pharmaceuticals. PMID- 10458735 TI - Monitoring of genes that respond to process-related stress in large-scale bioprocesses. AB - In large-scale aerobic fed-batch processes, cells are exposed to local zones of high glucose concentrations that can also cause local oxygen limitations at high cell densities. The mRNA levels of four stress genes (clpB, dnaK, uspA, and proU) and three genes responding to oxygen limitation or glucose excess (pfl, frd, and ackA) were investigated in an industrial 20-m(3) Escherichia coli process and in a scale-down reactor with defined high-glucose and low-oxygen zones. The mRNA levels of ackA and proU were high during the batch growth phase, but declined drastically when glucose became limited, whereas the mRNA levels of the other stress genes were relatively constant throughout the process. In the industrial scale reactor, the stress gene mRNA levels were, in most cases, highest in the middle part and at the top of the reactor, where the substrate was fed. Cells passing through the high glucose zone of the scale-down reactor had elevated mRNA levels for the oxygen limitation genes and had also elevated heat-shock gene mRNA levels. Both responses to stress occurred within seconds. The approach presented in this study offers a tool for monitoring process-related changes in the transcriptional regulation of genes. PMID- 10458736 TI - Expression of chimeric monomer and dimer proteins on the plasma membrane of mammalian cells. AB - Targeting of proteins to the plasma membrane of cells may be useful for vaccine development, tissue engineering, genetic research, bioseparations, and disease treatment. The ability of different transmembrane domains (TM) to direct a reporter protein (human alpha-feto protein, AFP) to the surface of mammalian cells was examined. High surface expression was achieved with chimeric proteins composed of AFP and the TM and cytosolic tail of murine B7-1 (AFP-B7) as well as with AFP containing a GPI-anchor from decay-accelerating factor (AFP-DAF). Lower surface expression of AFP was observed when the TM of human platelet-derived growth factor receptor or the human asialoglycoprotein receptor H1 subunit were employed. Introduction of the hinge-CH2-CH3 region of human IgG (gamma1 domain) between AFP and TM allowed efficient formation of disulfide-linked dimers. Surface expression of AFP-gamma1-B7 dimers was impaired compared to AFP-B7 whereas AFP-gamma1-DAF dimers were efficiently targeted to the surface. Accumulation of chimeric proteins on the cell surface did not correlate with the level of protein expression. This study demonstrates that high levels of monomeric and dimeric proteins can be targeted to the cell membrane of mammalian cells by proper selection of TM. PMID- 10458737 TI - Kinetic analysis of deactivation of immobilized alpha-chymotrypsin by water miscible organic solvent in kyotorphin synthesis. AB - Two different immobilized chymotrypsin derivatives were used to synthesize kyotorphin, using N-benzoyl-L-tyrosine ethyl ester and L-arginine ethyl ester as substrates, in water-DMF media. The first was adsorbed onto Celite particles and the second was multipoint covalently attached into polyacrylamide gel. In all cases, the conversion of the carboxyl substrate was carried out in first-order reaction conditions. For the adsorbed enzyme, the reaction kinetics deviated from first-order likely due to a fast irreversible inactivation of enzyme during the reaction time even at low DMF concentration (15-20% v/v). The covalent attachment of enzyme resulted in elimination of irreversible activity loss by organic solvent up to 60% (v/v) of DMF. The catalytic activity of the covalent derivative was conserved as appropriate for performing a synthetic reaction up to 60% v/v of DMF (in comparison to 30% v/v for the adsorbed derivative), showing a clear improvement in its stability against reversible denaturation by this solvent. The selectivity of the synthetic reaction was slightly enhanced (from 40-50%) with the increase in DMF concentration to 80% v/v, but it was significantly improved (to 80%) when L-argininamide was used as nucleophile. PMID- 10458738 TI - Slow heat rate increases yeast thermotolerance by maintaining plasma membrane integrity. AB - Thermal resistance of Saccharomyces cerevisiae was found to be drastically dependent on the kinetics of heat perturbation. Yeasts were found to be more resistant to a plateau of 1 h at 50 degrees C after a slope of temperature increase (slow and linear temperature increments) than after a shock (sudden temperature change). Thermotolerance was mainly acquired between 40-50 degrees C during a heat slope, i.e., above the maximal temperature of growth. The death of the yeasts subjected to a heat shock might be related to the loss of membrane integrity: intracellular contents extrusion, i.e., membrane permeabilization, was found to precede cell death. However, the permeabilization did not precede cell death during a heat slope and, therefore, membrane permeabilization was a consequence rather than a cause of cell death. During a slow temperature increase, yeasts which remain viable may have time to adapt their plasma membrane and thus maintain membrane integrity. PMID- 10458739 TI - Bicarbonate concentration and osmolality are key determinants in the inhibition of CHO cell polysialylation under elevated pCO(2) or pH. AB - Accumulation of CO(2) in animal cell cultures can be a significant problem during scale-up and production of recombinant glycoprotein biopharmaceuticals. By examining the cell-surface polysialic acid (PSA) content, we show that elevated CO(2) partial pressure (pCO(2)) can alter protein glycosylation. PSA is a high molecular-weight polymer attached to several complex N-linked oligosaccharides on the neural cell adhesion molecule (NCAM), so that small changes in either core glycosylation or in polysialylation are amplified and easily measured. Flow cytometric analysis revealed that PSA levels on Chinese hamster ovary (CHO) cells decrease with increasing pCO(2) in a dose-dependent manner, independent of any change in NCAM content. The results are highly pH-dependent, with a greater decrease in PSA at higher pH. By manipulating medium pH and pCO(2), we showed that decreases in PSA correlate well with bicarbonate concentration ([HCO(3)( )]). In fact, it was possible to offset a 60% decrease in PSA content at 120 mm Hg pCO(2) by decreasing the pH from 7.3 to 6.9, such that [HCO(3)(-)] was lowered to that of control (38 mm Hg pCO(2)). When the increase in osmolality associated with elevated [HCO(3)(-)] was offset by decreasing the basal medium [NaCl], elevated [HCO(3)(-)] still caused a decrease in PSA, although less extensive than without osmolality control. By increasing [NaCl], we show that hyperosmolality alone decreases PSA content, but to a lesser extent than for the same osmolality increase due to elevated [NaHCO(3)]. In conclusion, we demonstrate the importance of pH and pCO(2) interactions, and show that [HCO(3)(-)] and osmolality can account for the observed changes in PSA content over a wide range of pH and pCO(2) values. PMID- 10458740 TI - Cell lines with reduced UDP-N-acetylhexosamine pool in the presence of ammonium. AB - The glycosylation of pharmaglycoproteins from recombinant cell lines can be affected by an uncontrolled accumulation of ammonium in the medium. Glucosamine-6 phosphate isomerase (GPI) has been proposed as the key enzyme responsible for elevating the intracellular UDP-N-acetylhexosamine pool (UDPGNAc) by accepting ammonium from the medium of cultured mammalian cells. As previously reported, the increased UDPGNAc pool then affects the N-glycan complexity in glycoproteins. To understand the entry of extracellular ammonium into the cellular metabolism, GPI has been isolated to homogeneity from BHK-21 cells and characterized. Thus, the complete pathway by which ammonium enters the cellular metabolism was elucidated. To reduce the negative effects of ammonium, GPI was inhibited using two different strategies. First, the addition of mannose to the culture media and, second, antisense RNA expression. In both cases, the cellular UDPGNAc pool was suppressed in the presence of high ammonium concentrations in the medium. However, constant suppression of the UDPGNAc pool could not be achieved by antisense RNA expression because antisense clones were apparently unstable. Further studies showed that the main reason for instability was the inducibility of GPI by its substrate ammonium. GPI was induced to a factor of two under ammonium-containing medium conditions. We propose gene knockout technology for GPI repression to obtain cell lines consisting of an UDPGNAc pool unaffected by the presence of ammonium. PMID- 10458741 TI - GFP-expressing mammalian cells for fast, sensitive, noninvasive cell growth assessment in a kinetic mode. AB - This study correlates the fluorescent signal from stable recombinant CHO cell lines expressing the green fluorescent protein (GFP) at high levels with biomass or cell number, extending the use of fluorescent proteins to applications and assays where cell growth rates are important. Using a standard fluorometer, growth of these cells can be quantified noninvasively in multiwell plates, and because signals are obtained without preparation, the same culture samples can be measured repeatedly. Even with a small relative change in biomass, the specific growth rate can be determined in a few hours. The dynamics of cell populations can now be studied with high sensitivity, low error rate, and minimum sample preparation. PMID- 10458742 TI - Asymmetric synthesis of chiral amines with omega-transaminase. AB - The asymmetric synthesis of chiral amines using prochiral ketones was carried out with (S)-specific omega-transaminase (omega-TA) from Vibrio fluvialis JS17. This reaction is inhibited severely by both products, (S)-amine and deaminated ketone. In addition, thermodynamic equilibrium strongly favored the reverse reaction. L Alanine proved to be the best amino donor based on easy removal of the products. Optimal pH of the reactions with both whole cells and cell-free extract was 7. Amino acceptor reactivities of ketone substrates and reaction profiles of the asymmetric synthesis showed that the initial rate as well as the reaction yield were lower when the resulting (S)-amine from a prochiral ketone substrate was a more reactive amino donor. The yield could be increased dramatically by removing pyruvate, which is a more inhibitory product than (S)-alpha-methylbenzylamine [(S)-alpha-MBA] when acetophenone and L-alanine are used as an amino acceptor and donor, respectively. The removal of pyruvate was carried out by incorporating lactate dehydrogenase (LDH) in cell-free extract or by using whole cells. The whole cell reaction yielded a much better result. When 25 mM benzylacetone and 30 mM acetophenone were used as an amino acceptor with 300 mM L-alanine, 90.2% and 92.1% of the reaction yields after 1 day were obtained with whole cells, respectively. Enantiomeric excesses of both (S)-alpha-MBA and (S)-1-methyl-3 phenylpropylamine [(S)-MPPA] were all above 99%. PMID- 10458743 TI - Wine yeast fermentation vigor may be improved by elimination of recessive growth retarding alleles. AB - The presence of recessive growth-retarding alleles can reduce the fitness of industrial wine yeasts. In nature, these alleles are supposed to be eliminated through "genome renewal". We emulated this process in the laboratory to increase the fermentation vigor of wine yeasts. The procedure is simply to sporulate the yeast strains and select new homozygous single-spore descendants. Most of the yeasts achieve a faster onset of fermentation when recessive deleterious genes are eliminated. The increase of the degree of homozygosity has no relation, either direct or inverse, with the fermentation vigor of the yeasts or with the quality of the resulting wine. However, in some strains in which recessive growth retarding alleles have been eliminated, the fermentation vigor and the quality of the wine were found to be improved simultaneously. PMID- 10458744 TI - Novel strategy for efficient screening and construction of host/vector systems to overproduce penicillin acylase in Escherichia coli. AB - A novel and simple method of using penicillin for screening of mutant strains with a high penicillin acylase (PAC) activity was developed. Random mutagenesis was conducted using a PAC-producing strain resistant to 6-aminopenicillanic acid (6-APA) as the parent strain and mutants were screened with penicillin at a high concentration. Results suggest that mutants with a high minimum inhibitory concentration for penicillin (MIC(penG)) usually overproduce PAC. Both volumetric and specific PAC activities of a mutant, MD7, were significantly higher than those of the parent strain, HBPAC101 harboring pCLL2902. The mutation(s) resulting in the enhanced expression was mapped on the host chromosome rather than the plasmid. In addition, the mutant strain of MDDeltaP7, derived by elimination of the harbored plasmid in MD7, was demonstrated to be efficient in production of PAC by using the expression plasmids for which expression of the pac gene is limited by translation. An extremely high specific PAC activity of more than 350 U/L/OD(600) was reached upon cultivation of MDDeltaP7 harboring pTrcKnPAC2902 in a bioreactor. As such, the strategy is effective in terms of constructing PAC overproducers and improving the process yield for production of PAC. PMID- 10458745 TI - Enantioselective recognition mechanism of secondary alcohol by surfactant-coated lipases in nonaqueous media. AB - The enantioselective recognition mechanism of secondary alcohol by lipases originated from Candida rugosa and Pseudomonas cepacia was elucidated on the basis of the kinetic study of the esterification of alcohol with lauric acid in isooctane. To obtain inherent kinetic parameters, we utilized a surfactant-coated lipase whose conformation is considered to be an "open" form in a homogeneous organic solvent. Based on the experimental results, the enantioselectivity of lipases was found to be derived from the difference in the V(max) values between the two enantiomers. The same result was observed when lipases of different origin and substrates with different molecular structures were applied. © 1999 John Wiley & Sons, Inc. PMID- 10458746 TI - Construction of a multi RE module: exploitation of mechanochemistry of restriction endonucleases. AB - We describe the construction of a multi-immobilized restriction endonuclease module (Multi RE module). We demonstrate that the applied mechanical stress enables modulation of enzyme activity and modulation of recognition site selectivity (in oligonucleotides of approximately 200 bp) of immobilized restriction endonucleases. The central module which is consisted of different strips of immobilized restriction endonucleases allows limited digestion of a large DNA sample in a controlled manner as a function of applied mechanical stress on strips. The stress-activity relationship and the effect of repeated cycles of stress and relaxation on the immobilized strips are presented here. © 1999 John Wiley & Sons, Inc. PMID- 10458747 TI - Problems in predicting cell damage from bubble bursting. AB - The question is addressed as to whether observed parameter(s) characterizing single bubble burst (bubble jet height and speed) can be used to predict cell damage in sparged animal cell cultures. Bubble burst profiles are examined in the presence of realistic concentrations of fetal calf serum (FCS) or Pluronic F-68 using a high-speed video technique. The damage to TBC3 hybridoma cells from bubble sparging, characterized as a first-order decline, is reduced by even very small concentrations of both FCS and Pluronic F-68, but neither single bubble burst parameters nor surface properties give usable correlations with death rate constants. © 1999 John Wiley & Sons, Inc. PMID- 10458748 TI - Blocking CD40 - CD154 and CD80/CD86 - CD28 interactions during primary allogeneic stimulation results in T cell anergy and high IL-10 production. AB - Although CD28 triggering provides an important co-stimulatory signal to T cells, blocking the CD80/CD86 - CD28 interaction with CTLA-4lg fusion protein is not sufficient for tolerance induction in vivo or in vitro. According to more recent data, interruption of the CD40 - CD154 interaction might complement the effect of CTLA-4lg and induce graft acceptance. We studied the effects of a blocking anti CD40 monoclonal antibody (mAb) and/or blocking anti-CD80/anti-CD86 mAb in cultures of human peripheral blood mononuclear cells (PBMC) stimulated with allogeneic PBMC. T cells activated by alloantigens in the presence of anti-CD80, anti-CD86 and anti-CD40 entered a state of alloantigen-specific non responsiveness as evidenced upon restimulation by lack of proliferation, cytotoxic activity, and IL-2, IL-5 and IL-13 production. IFN-gamma production during restimulation was less than in the control cultures, while the production of IL-10 was enhanced. Addition of recombinant IL-2 during the restimulation rescued alloantigen-specific activity. We conclude that the simultaneous blocking of the CD40 - CD154 and CD80/CD86 - CD28 interaction during allogeneic T cell activation induces T cell anergy. Since anergic cells induced by this treatment still produce high levels of IL-10, the latter could contribute to modulation of antigen-presenting cell activity and to bystander suppression of residual reactive T cells. PMID- 10458749 TI - The CD45 tyrosine phosphatase regulates CD3-induced signal transduction and T cell development in recombinase-deficient mice: restoration of pre-TCR function by active p56(lck). AB - The pre-TCR complex regulates the transition from CD4(-)CD8(-) double-negative (DN) to CD4(+)CD8(+) double-positive (DP) thymocytes during T cell development. In CD45(-/-) mice there is an accumulation of DN cells, suggesting a possible role for CD45 in pre-TCR signaling. We therefore crossed CD45(-/-) with Rag-1(-/ ) mice to investigate the signaling functions of the CD3 complex in DN thymocytes. Remarkably, treatment of Rag-1(-/-)/CD45(-/-) mice with a CD3 mAb caused maturation to the DP stage at only 3% of the level measured in Rag-1(-/-) mice. Furthermore, ligation of the CD3 complex on Rag-1(-/-) /CD45(-/-) thymocytes in vitro induced less tyrosine phosphorylation in specific proteins when compared to Rag-1(-/-) thymocytes. CD45(-/-) mice were also crossed with pLGFA mice expressing a constitutively active form of the lck tyrosine kinase which restored the DN to DP transition to near normal levels. Our results are consistent with a model in which CD45-activated p56(lck) is critical for pre-TCR signal transduction. PMID- 10458750 TI - Quantitative determination of TCR cross-reactivity using peptide libraries and protein databases. AB - A single T cell clone can be activated by many different peptides in the context of a particular HLA molecule. To quantify the number of peptides that can be recognized by a CD4(+) T cell clone, we screened a one-bead-one-peptide synthetic peptide library and a protein database for peptides that stimulate an HLA-DR3 restricted, human glutamic acid decarboxylase (GAD65)-reactive CD4(+) T cell clone. Both the library screening and the database analysis indicated that this T cell clone is able to recognize approximately 10(6) 11-mer peptides at low nanomolar concentration. Furthermore, we determined that the frequency of cross reactivity increased only 1.5-3 times when the peptide concentration increased 10 times, in the range of 0.01 - 1 microM. These data imply that there is a considerable potential for T cell cross-reactivity and are useful for studies on the role of molecular mimicry in the etiology of T cell-mediated disease. PMID- 10458751 TI - Gene structure of the murine NK cell receptor 2B4: presence of two alternatively spliced isoforms with distinct cytoplasmic domains. AB - The NK cell receptor 2B4 is expressed on the surface of all murine NK cells and a subset of T cells. Ligation of 2B4 with monoclonal antibodies increases target cell lysis and IFN-gamma production. 2B4 is the high-affinity counter-receptor for CD48 in mice and humans. 2B4-L is a member of the CD2 subgroup of the immunoglobulin supergene family, which includes CD48, LFA-3, CD84, Ly9 and SLAM. Here we describe 2B4-S, a second 2B4 isoform, and the genomic structure of the 2B4 gene. 2B4-S is identical to the 5' end of 2B4-L, differing only at the 3' end, corresponding to a portion of the cytoplasmic domain and the 3' untranslated sequence. Both 2B4-L and 2B4-S are expressed on IL-2-activated NK cells. The genomic clone of 2B4 reveals that the two cDNA clones are products of alternative splicing. Since they differ only in a portion of the cytoplasmic domain, it is likely that they transduce different signals. PMID- 10458752 TI - Outcome of Staphylococcus aureus-triggered sepsis and arthritis in IL-4-deficient mice depends on the genetic background of the host. AB - Disruption of the IL-4 gene in two inbred mouse strains revealed a dual role of IL-4 in Staphylococcus aureus sepsis and arthritis depending on the host's genetic background. IL-4 was protective in 129SV mice, since 5 days after S. aureus inoculation IL-4(-/-) mice displayed 70% mortality as compared to survival of all 129SV wild-type counterparts. On the other hand, IL-4 was detrimental in C57BL/6 mice, since survival of IL-4(-/-) C57BL/6 mice was increased, as compared to wild-type controls, due to decreased staphylococcal growth. Altogether, our results show the dual role of IL-4 in S. aureus sepsis and arthritis, depending on the genetic background of the host. PMID- 10458753 TI - CD8(+)NKR-P1A (+)T cells preferentially accumulate in human liver. AB - A unique subset of T cells that co-express NKR-P1, which is a lectin type of NK receptor and is thought to have a major role in triggering NK activity, has been identified. In mice, NK1.1 (mouse NKR-P1C)(+) T cells, called NKT cells, preferentially accumulate in the liver and bone marrow. They predominantly use invariant Valpha14 chain TCR and phenotypically are CD4(+)CD8(-) or CD4(-)CD8(-) T cells. In this study, we analyzed, phenotypically and functionally, the NKR-P1A (analogue of murine NKR-P1C)(+) T cells resident in the human liver. Here, we show that in complete contrast to the NKT cells in the mouse liver, the majority of NKR-P1A(+) T cells in the human liver are CD8(+) and their TCR repertoire is not skewed to Valpha24 TCR, the homologue of murine Valpha14 TCR. Almost all of the NKR-P1A(+) T cells in the human liver expressed CD69, suggesting that they were activated. Furthermore, the NKR-P1A(+) T cells in the human liver exhibited strong cytotoxicity against a variety of tumor cell lines including K562, Molt4 and some colonic adenocarcinoma cell lines. PMID- 10458754 TI - The role of calcium homeostasis and flux during bacterial antigen processing in murine macrophages. AB - We report that MHC class II (MHC-II)-restricted antigen processing of two CD4(+) T cell epitopes from the surface M protein of Streptococcus pyogenes in murine macrophages is dependent on intact calcium homeostasis and flux. We have previously shown that the CD4(+) T cell epitope 308-319 of the type 5 M protein is presented by newly synthesized MHC-II molecules via the classical pathway, while 17-31 is loaded on recycling MHC-II molecules via the recycling pathway. In this report we show that processing of viable bacteria for 308-319 presentation depended on the availability of intra- and extra cellular calcium, intact gadolinium-sensitive and/or T-type calcium channels, as well as on thapsigargin sensitive homeostasis of intracellular calcium. In contrast, processing of 17-31 was independent of both intracellular calcium and gadolinium-sensitive calcium channels. The data suggest that alternative antigen processing pathways have different requirements for intracellular calcium homeostasis. PMID- 10458755 TI - Exceptionally long CDR3H region with multiple cysteine residues in functional bovine IgM antibodies. AB - We analyzed VDJ and VJ rearrangements in IgM-secreting B lymphocytes from a cow infected with bovine leukemia virus (BLV). BLV causes expansion of CD5(+) and IgM(+) B lymphocytes regardless of antigen specificity. The data showed that single point mutations contribute to the diversification of IgM antibodies. The most striking observation, however, is that approximately 9% of theVDJ rearrangement in IgM-secreting B cells encode an exceptionally long third complementarity-determining region of the heavy chain (CDR3H; 56 to 61 amino acids) with multiple cysteine residues. Such an exceptionally long CDR3H is the first ever to be documented for an antibody in a species. These VDJ rearrangements encode functional IgM antibodies as some of these show polyspecific reactivity. The presence of even-numbered cysteine residues in the CDR3H may provide hitherto unknown configurational ability to the antigen combining site via intra-CDR3H disulfide bridging. In addition, the VDJ rearrangements encoding exceptionally long CDR3H paired with either novel V(lambda)1 or V(x)1x genes, earlier noted not to be expressed. Overall, these experiments provide evidence that somatic hypermutations and generation of an exceptionally long CDR3H contribute to the diversification of IgM antibodies in cattle. PMID- 10458756 TI - Induction of inhibitory antibodies to the CCR5 chemokine receptor and their complementary role in preventing SIV infection in macaques. AB - The seven-transmembrane G-protein-linked CCR5 molecule functions as a major coreceptor for HIV or simian immunodeficiency virus (SIV) infection. Antibodies to CCR5 were studied in rhesus macaques immunized with SIV grown in human CD4(+) T cells. These macaques were completely protected against i.v. challenge with live SIV. Sera from the protected macaques showed significantly greater inhibition of SIV replication (p < 0.001) and macrophage inflammatory protein 1beta-generated CCR5-dependent chemotaxis (p < 0.01) than sera from unprotected macaques, in the absence of significant neutralizing antibodies to SIV. These two functional assays demonstrate serum antibodies to the CCR5 receptors which were specifically inhibited by CCR5-transfected HEK-293 cells. We postulate that anti CCR5 antibodies may be complementary to beta-chemokines in blocking CCR5 coreceptors to HIV or SIV binding and fusion of CD4(+) cells. PMID- 10458757 TI - Normal human immunoglobulin suppresses experimental myasthenia gravis in SCID mice. AB - Serum IgM has been shown to participate in the control of IgG autoreactivity in healthy subjects. We have recently shown that an immunoglobulin preparation of pooled normal human IgM (IVIgM) contains anti-idiotypic antibodies against disease-associated IgG autoantibodies in autoimmune patients and protects rats from experimental autoimmunity. The aim of the present study was to asses the in vitro and in vivo immunomodulatory effects of IVIgM in comparison with IgG, in SCID mice reconstituted with thymic cells from a myasthenia gravis patient. Non leaky SCID mice were injected i.p. with 60 x 10(6) thymic cells from a patient with myasthenia gravis and 1 day later boosted with 10(6) irradiated acetylcholine receptor (AchR)-expressing TE671 cells. On days 14, 21 and 28, mice were treated with IVIgM or with equimolar amounts of human serum albumin. The level of anti-AchR antibodies in the sera of three out of four IgM-treated animals was less than 1 nM. Further, there was a significant decrease in the loss of endplate AchR on the diaphragms of IgM-treated SCID mice. These findings indicate that pooled normal IgM exerts an immunoregulatory role in experimental myasthenia gravis, and suggests that IgM may be considered as an alternative approach in the therapy of autommune diseases. PMID- 10458758 TI - CD28 co-stimulation results in down-regulation of lymphotactin expression in human CD4(+) but not CD8(+) T cells via an IL-2-dependent mechanism. AB - Chemokines are key molecules in promoting leukocyte migration and, for some of them, T cell adhesion and activation. Lymphotactin, which is the unique known member of the C class of chemokines, is produced by and acts on T lymphocytes, but the requirement of co-stimulatory pathways such as CD28 for its expression is largely unknown. CD28 plays a dominant role in the amplification of T cell proliferation, survival and cytokine production. In this report, we demonstrate that human lymphotactin expression, at both the mRNA and protein levels, is optimally induced by CD3/TCR activation alone, whereas CD28 co-stimulation turns off this expression. This down-regulation is not attributable to secondary activation via CTLA-4, the alternative counter-receptor of B7 ligands. Only the CD4(+) and not the CD8(+) subset is directly affected by this negative regulation. Transcript destabilization can be ruled out as a mechanism by which CD28 down-regulates lymphotactin expression. However, such down-regulation can be partly induced by IL-2 and abrogated by blocking IL-2/IL-2 receptor interaction. This particular profile of lymphotactin expression is not in line with the prevailing dogma of up-regulation of cytokine gene expression by CD28 co stimulation, and represents a new CD28-mediated regulatory mechanism for lymphotactin expression. PMID- 10458759 TI - SSTR2A is the dominant somatostatin receptor subtype expressed by inflammatory cells, is widely expressed and directly regulates T cell IFN-gamma release. AB - Macrophages secrete the immunoregulatory peptide somatostatin (SOM) that inhibits IFN-gamma release by splenocytes and granuloma cells of schistosome-infected mice. In this report we demonstrate that granuloma cells express mRNA for the SOM receptor SSTR2 but not the other four SSTR subtypes. Blocking SSTR2 activity with anti-SSTR2 antiserum prevents SOM inhibition of T cell IFN-gamma production. This demonstrates that SOM regulates T cell function via SSTR2. Two isoforms of SSTR2 exist due to alternative RNA splicing. We developed sensitive and specific competitive PCR assays to quantify total SSTR2, SSTR2A and SSTR2B mRNA levels. The SSTR2A isoform accounts for 99% of inflammatory cell SSTR2 mRNA and does not appear to be regulated at the transcripitonal level. B cells and macrophage cell lines also express SSTR2 mRNA which raises the possibility that SOM influences T cell IFN-gamma release by regulating accessory cell function. We show that SOM acts directly on T cells to inhibit TCR-stimulated IFN-gamma release. Thus, SOM may directly regulate T cell IFN-gamma release at inflammatory sites. PMID- 10458760 TI - Effect on parasite eradication of Pneumocystis carinii-specific antibodies produced in the presence or absence of CD4(+) alphabeta T lymphocytes. AB - The contribution of specific antibodies (Ab) to successful clearance of Pneumocystis carinii from host pulmonary tissues has received increasing attention. Sera collected from diseased recombinase-activating gene (RAG)-1(-/-), TCRbetaxdelta(-/-), TCRbeta(-/-) and Abeta(-/-) mutants as well as from aerogenic parasite-exposed (aero) and intranasally (i. n.) infected C57BL/6 mice were transferred to RAG-1(-/-) mutants inoculated with freshly isolated parasites. All sera, except for RAG-1(-/-) serum, contained P. carinii-specific Ab of varying isotype concentrations. Four weeks after serum treatment pulmonary parasite numbers were reduced slightly by Abeta(-/-) and C57BL/6-aero sera, and markedly by TCRbeta(-/-) and C57BL/6-i.n. sera. Our data reveal: (1) T cells are essential, and CD4(+) T cells are important for formation of protective Ab; (2) at least in the absence of alpha beta T cells, gamma delta T cells provide help for protective Ab. In vitro treatment of bronchoalveolar lavage cells with the different sera largely led to comparable results. Opsonizing Ab impeding parasite attachment to host cells, as well as Ab possibly neutralizing parasite-secreted products were implicated. Furthermore, serum components other than Abappear to participate in resistance to fungal manifestation. PMID- 10458761 TI - Molecular and cellular aspects of induced thymus development in recombinase deficient mice. AB - Thymus development and microenvironment organization require stage- and site specific cross-talk between thymocyte and stroma. In this study we have used recombinase-activating gene-deficient (RAG-2(-/-)) mice to analyze regulated gene expression both in thymocytes and stromal cells following injection of anti-CD3 monoclonal antibodies as inducer of thymus development. We show that IFN-gamma, TNF-alpha and lymphotactin are transcriptionally regulated in thymocytes, whereas cytoskeletal keratin 14, IL-1alpha and TNF-alpha are regulated in the stroma, quantitatively reproducing the variations associated with beta selection of thymocytes. In addition, RAG-2(-/-) thymus development is associated with entry of epithelial cells into the cell cycle. The histochemical evidence that expanded RAG-2(-/-) thymus becomes undistinguishable from wild-type cortex further suggests that cross-talk phenomena occurring during beta selection of thymocyte are reproduced in this system. PMID- 10458762 TI - Regulation of cell survival during B lymphopoiesis: suppressed apoptosis of pro-B cells in P53-deficient mouse bone marrow. AB - B cell development in mouse bone marrow (BM) is subject to quality controls that eliminate aberrant cells by apoptosis, but the intrinsic cellular mechanisms that mediate this negative B cell selection remain unclear. The p53 tumor suppressor transduces signals resulting in apoptosis and cell cycle arrest in cells that sustain DNA damage. Faulty V(D)J recombination in scid lymphocyte precursors activates a p53-dependent DNA damage checkpoint. In the present study, we have examined whether p53 is involved in apoptotic selection of normally developing B cells in BM. Double immunofluorescence labeling and flow cytometry were used to quantitate phenotypically defined B cell populations and their apoptotic rates in BM of homozygous p53-deficient mice. B220(+) mu(-) and terminal deoxynucleotidyl transferase (TdT)(+) pro-B cells were increased in both incidence and absolute number to controls. In contrast, pre-B cells were only slightly increased and the sIgM(+) B lymphocyte compartment remained essentially normal. The incidence of apoptosis among p53(-/-) pro-B cells was greatly reduced, both ex vivo and in short-term culture, whereas, apoptosis of pre-B cells and B lymphocytes was not significantly different from normal. The results indicate that p53 is actively involved as an apoptosis inducer at an early quality control checkpoint in B lymphopoiesis. PMID- 10458763 TI - An airbone mold-derived product, beta-1,3-D-glucan, potentiates airway allergic responses. AB - Repeated inhalation of allergen leads to the down-regulation of allergen-specific IgE responses in non-atopic individuals as well as in mice. This phenomenon is named inhalation-induced IgE tolerance. In contrast, inhaled allergen causes significant IgE and allergic responses in atopic persons. The mechanisms involved in this differential regulation of airway allergen-specific immune responses remain unclear. Besides the allergen exposure of genetically susceptible individuals, environmental contamination is considered to play a role as an initiating factor for airway allergic responses. Using a murine model, we demonstrate here that airborne beta-1, 3-D-glucan, which exists frequently in our environment, particularly in highly humid areas, can abrogate inhalation-induced IgE isotype-specific down-regulation and promote airway eosinophil infiltration to inhaled antigen. PMID- 10458764 TI - Effects of nitric oxide on the induction and differentiation of Th1 cells. AB - We have previously shown that mice lacking inducible NO synthase are markedly more susceptible to Leishmania major infection but developed a significantly enhanced Th1 cell response compared with wild-type mice. Furthermore, at high concentrations, NO inhibited IL-12 synthesis by activated macrophages, thereby indirectly suppressing the expansion of Th1 cells. We report here that at low concentrations, NO selectively enhanced the induction of Th1 cells and had no effect on Th2 cells. NO exerted this effect in synergy with IL-12 during Th1 cell differentiation and had no effect on fully committed Th1 cells. NO appears to affect CD4(+) T cells directly and not at the antigen-presenting cells. These results therefore provide an additional pathway by which NO modulates the immune response and contributes to the homeostasis of the immune system. PMID- 10458765 TI - HLA binding and T cell recognition of a tissue transglutaminase-modified gliadin epitope. AB - DQ2 confers susceptibility to celiac disease (CD) and intestinal CD4(+) T cells of DQ2(+) CD patients preferentially recognize deamidated gliadin peptides. This modification can be mediated by tissue transglutaminase (tTG). We have investigated what role the tTG-modified residues play in DQ2 binding and T cell presentation using a model gamma-gliadin peptide (residues 134 - 153). Treatment of this peptide with tTG resulted in deamidation of Gln residues at positions 140, 148 and 150. Two of these residues act as DQ2 anchors at position P7 (148) and P9 (150) and increased the affinity of the modified peptide for DQ2 50-fold. Testing of a mutant DQ2 molecule demonstrated that the Lys residue at beta71 of DQ2 is important for binding of the deamidated peptide. A variant DQ2 molecule (with the same beta-chain but different alpha-chain) that does not confer susceptibility to CD was capable of presenting the gliadin peptide, but not pepsin/trypsin-digested gliadin, equally well to a T cell. This suggests that processing events might be involved in the preferential presentation of the gliadin peptide by the DQ2 molecule. Substitution of Gln with Glu in some positions not targeted by tTG, but in positions likely to be deamidated via non enzymatic mechanisms, disrupted T cell recognition. This provides additional evidence that tTG is responsible for modification of gliadin in vivo. PMID- 10458766 TI - Identification and function of neonatal Fc receptor in mammary gland of lactating mice. AB - In addition to its proposed function in regulating serum IgG levels, the MHC class I-related neonatal Fc receptor (FcRn) is known to play a role in IgG transfer across rodent yolk sac and neonatal intestine. In contrast to humans, for which transplacental transfer of IgG appears to be the only mechanism of maternal IgG delivery, the transmission of IgG in mice occurs both antenatally (yolk sac) and neonatally (transport from mother's milk across intestinal epithelial cells). In the current study, a possible role for FcRn in regulating IgG transfer into milk has been investigated. FcRn has been shown to be present in functional form in the mammary gland of lactating mice, and is localized to the epithelial cells of the acini. Analysis of the transfer of Fc fragments and IgG which have different affinities for FcRn indicate that, unexpectedly, these proteins are transferred in inverse correlation with their binding affinity for FcRn. Thus, in the lactating mammary gland FcRn appears to play a role in recycling IgG in a mode that may have relevance to FcRn trafficking during the maintenance of constant serum IgG levels. PMID- 10458768 TI - The keratinocyte-derived cytokine IL-7 increases adhesion of the epidermal T cell subset to the skin basement membrane protein laminin-5. AB - Human epidermis contains a subset of epidermal T cells that can mount an immune response by migrating through the skin and into the peripheral lymphnodes to proliferate before re-entering the epidermis. The cytokine IL-7 is shown to be localized to the basement membrane of normal human skin. Furthermore, culturing in the presence of IL-7 causes increased adhesion of epidermal T cells but not peripheral blood T cells to the major epidermal basement membrane protein, laminin-5. The mechanism for increased T cell adhesion to laminin-5 is due, at least in part, to an increase in the cell surface expression of the integrin alpha3beta1. Epidermal T cells cultured in IL-7 that are strongly adherent to laminin-5 are shown by flow cytometry to consist of a variety of subsets; therefore, the increase in cell adhesion is not due to an outgrowth of one T cell subset during culturing. We hypothesize that in vivo, exposure to IL-7 is required for epidermal T cell adhesion to laminin-5. PMID- 10458767 TI - STAT-4 mediated IL-12 signaling pathway is critical for the development of protective immunity in cutaneous leishmaniasis. AB - Recent studies have demonstrated that two IL-12 signaling pathways, a STAT 4 - dependent and STAT4 - independent, are involved in the development of a Th1-like response. To determine their roles in the development of protective immunity against Leishmania major, we monitored progression of cutaneous Leishmania major infection in STAT4-deficient mice (STAT4-/-) compared to similarly infected wild type (STAT4+/+) mice. Although the onset of lesion growth was delayed in STAT4-/- mice during the early phase of infection, these mice eventually developed large, non-healing lesions, whereas STAT4+/+ mice resolved their lesions. As infection progressed, both STAT4+/+ and STAT4-/- mice infected with L. major displayed similar titers of Leishmania-specific IgG1 and IgE but later produced lower IgG2a. On days 20 and 40 post-infection, Leishmania antigen-stimulated lymphnode cells from STAT4-/- mice produced significantly lower amounts of IFN-gamma than those from STAT4+/+ mice as measured by enzyme-linked immunosorbent assay. There was no significant difference, however, in IL-4 and IL-12 production between the two groups. These results indicate that STAT4-mediated IL-12 signaling is critical for the development of protective Th1 response following L. major infection in genetically resistant mice. Additionally, they demonstrate that, although genetically resistant mice lacking STAT4 signaling pathway develop large, non-healing lesions, they do not default towards a Th2-like response. PMID- 10458769 TI - Dephosphorylation of ZAP-70 and inhibition of T cell activation by activated SHP1. AB - Studies with motheaten mice, which lack the SHP1 protein tyrosine phosphatase, indicate that this enzyme plays an important negative role in T cell antigen receptor (TCR) signaling. The physiological substrates for SHP1 in T lymphocytes, however, have remained unclear or controversial. To define these targets for SHP1 we have compared the effects of constitutively active and inactive mutants of SHP1 on TCR signaling. Expression of wild-type SHP1 had a very small effect on the TCR-induced tyrosine phosphorylation of ZAP-70 and Syk, even when SHP1 was overexpressed 20 - 100-fold over endogenous SHP1. Inactive SHP1-D421A and wild type SHP2 were without effects. Constitutively active SHP1-DeltaSH2 had a more pronounced effect on ZAP-70 and Syk, even when expressed at near physiological levels. SHP1-DeltaSH2 also inhibited events downstream of ZAP-70 and Syk, such as activation of the mitogen-activated protein kinase Erk2 and the transcriptional activation of the interleukin-2 gene. In contrast, a constitutively active SHP2 DeltaSH2 had no statistically significant effect (although it caused a slight augmentation in some individual experiments). None of the constructs influenced the anti-CD3-induced tyrosine phosphorylation of the TCR zeta-chain or phospholipase Cgamma1, indicating that Src family kinase function was intact. Taken together, our findings support the notion that ZAP-70 and Syk can be direct substrates for SHP1 in intact cells. However, the two SH2 domains of SHP1 did not facilitate its recognition of ZAP-70 and Syk as substrates in intact cells. Therefore, we suggest that SHP1 is not actively recruited to inhibit TCR signaling induced by ligation of this receptor alone. Instead, we propose that ligation of a distinct inhibitory receptor leads to the recruitment of SHP1 via its SH2 domains, activation of SHP1 and subsequently inhibition of TCR signals if the inhibitory receptor is juxtaposed to the TCR. PMID- 10458770 TI - Fractalkine, a CX3C chemokine, is expressed by dendritic cells and is up regulated upon dendritic cell maturation. AB - The lone CX3C chemokine, fractalkine (FK), is expressed in a membrane-bound form on activated endothelial cells and mediates attachment and firm adhesion of T cells, monocytes and NK cells. We now show that FK is associated with dendritic cells (DC) in epidermis and lymphoid organs. In normal human skin, dual-color fluorescence microscopy co-localized FK expression with Langerhans cells expressing CD1a. In tonsil, FK-positive DC expressed CD83, a marker for mature DC. Human and murine cultured DC up-regulated FK mRNA expression with maturation. Furthermore, CD40 ligation, but not TNF-alpha or lipopolysaccharide treatment, of activated, migratory DC that had migrated from skin explants resulted in a 2.5 fold increase of surface expression of FK without significant alterations of expression of CD80, CD86, CD54 or MHC class II. Since FK mediates adhesion of T cells to activated endothelial cells, the increased expression of FK during DC maturation (and particularly by CD40 ligation) may play a role in the ability of T cells and mature DC to form conjugates and engage in cell-cell communication. PMID- 10458771 TI - Inhibition of experimental autoimmune encephalomyelitis in SJL mice by oral administration of retro-inverso derivative of encephalitogenic epitope P87-99. AB - Retro-inverso modification of peptides preserves parent peptide overall topology and provides at the same time stability to proteolysis, leading to derivatives with prolonged half-life in vitro and in vivo. In this study the encephalitogenic epitope P87 - 99 of myelin basic protein has been prepared in the retro-inverso form to examine its biological activity in a murine model of multiple sclerosis. Experiments of in vivo T cell tolerance induction in SJL mice revealed that the retro-inverso peptide was able to induce a selective T cell hyporesponsiveness, as measured by a reduction in the proliferative response of lymphnode T cells after antigen challenge. Oral administration of retro-inverso peptide decreased the disease severity significantly and delayed considerably the disease onset in treated mice. Enhancement of resistance to proteolysis by retro-inverso modification of encephalitogenic epitopes may increase the therapeutic value of oral tolerance induction in the treatment of multiple sclerosis and other Th1 associated inflammatory disorders. PMID- 10458772 TI - Human acute myeloblastic leukemia cells differentiate in vitro into mature dendritic cells and induce the differentiation of cytotoxic T cells against autologous leukemias. AB - An immune response is involved in the control of leukemias as demonstrated by allogeneic bone marrow transplantation, by the eradication of residual leukemic cells by cytotoxic T cells and finally by the identification of tumor antigens which are recognized by effector T cells. Dendritic cells (DC) are professional antigen-presenting cells (APC) able to present antigens in the context of co stimulatory signals necessary for T cell activation. Although tumor cells may express tumor antigens, they are usually unable to elicit an immune response since they are devoid of co-stimulatory capacities. To overcome this problem, engineering tumors to provide APC function could potentially result in polyvalent immunization to multiple tumor antigens. We have tested the differentiation of AML-5 (monoblastic, promonocytic and monocytic) leukemia cells and demonstrated that eight out of the ten fresh human acute myeloid leukemia populations tested can differentiate in vitro into bona fide APC. Leukemic cells acquire in vitro DC morphology, mature DC markers such as CD83, the up-regulation of MHC and co stimulatory molecules and the ability to produce IL-12 upon maturation, while retaining their characteristic caryotypic abnormalities. However, we could not obtain an immature DC phenotype. They also acquire the ability to induce the differentiation of allogeneic naive cord blood CD4 and CD8 T cells as well as resting autologous cytotoxic T cells. These results demonstrate that some tumor cells acquire APC phenotype and functions and can thereby induce a potent autologous immune response that will be a valuable tool for detection of new tumor antigens and for in vivo immunization. PMID- 10458773 TI - Selective amino acid substitutions of a subdominant Epstein-Barr virus LMP2 derived epitope increase HLA/peptide complex stability and immunogenicity: implications for immunotherapy of Epstein-Barr virus-associated malignancies. AB - The latent membrane protein 2 is an immunogenic antigen expressed in Epstein-Barr virus (EBV)-associated tumors and consequently it may represent a target for specific cytotoxic T lymphocyte (CTL)-based immunotherapies. However, the efficacy of such a therapy is limited by the poor immunogenicity of the protein that induces weak CTL responses directed to the CLGGLLTMV (CLG) epitope only in the minority of EBV-seropositive donors. We have now demonstrated that selective peptide stimulation of peripheral blood lymphocytes induced CLG-specific CTL in all donors, suggesting that this epitope can be a suitable target for specific immunotherapies. We found that the CLG peptide has a low affinity for HLA-A*0201 and does not produce stable complexes, both factors that are likely to determine the strength of CTL responses to this epitope. Therefore, we synthesized and tested CLG analogues carrying single or combined amino acid substitutions to increase HLA/peptide stability. Among the analogues tested we identified two peptides which, compared to the natural epitope, showed higher affinity for HLA A*0201 molecules, and produced stable complexes. These peptides demonstrated a potent, specific stimulatory capacity and could be used for selective CTL-based therapies. PMID- 10458774 TI - Direct evidence for native CD4 oligomers in lymphoid and monocytoid cells. AB - CD4 is expressed by T lymphocytes and monocytes and is generally considered a monomer even though its structure was originally modelled on the REI Bence-Jones homodimer. However, native CD4 was demonstrated as both monomer and dimers of 55 and 110 kDa in lymphoid and monocytoid cells by immunoprecipitation and immunoblotting after solubilization with alkylating (iodoacetamide) or reducing (dithiothreitol, 2-mercaptoethanol) reagents. Full reduction yielded only the 55 kDa monomeric form. Purified CD4 oligomers from CEM-T4 cells were also resolved as homodimers by MALDI-Tof mass fingerprinting after tryptic digestion. Cell treatment with the membrane impermeable, free-thiol reactive, 5,5'-dithiobis-2 nitrobenzoic acid enhanced cell surface CD4 dimers and tetramers. The interaction sites producing dimerization were probably in the D4 domain as OKT4 inhibited self association of recombinant CD4 (rCD4). Oligomerization of rCD4 by glutathione and thioredoxin indicates that thiol exchange interactions were responsible. Enhanced CD4 dimer expression was also observed after PMA (20 ng/ml) activation of THP-1 cells. These findings demonstrate that different quaternary forms of CD4 such as monomers, homodimers and tetramers are expressed by T lymphocytes and monocytes/macrophages. PMID- 10458775 TI - Inhibition of T cell apoptosis by IFN-beta rapidly reverses nuclear translocation of protein kinase C-delta. AB - Type I interferons rescue activated human T cells from cytokine deprivation induced apoptosis. Our data now show that IFN-beta also rapidly inhibits apoptotic signals induced through the Fas receptor (CD95) in human T cells. To identify upstream signaling elements that could be targets of IFN-beta, we have studied protein kinase C (PKC). PKC-delta is actively involved in the regulation of apoptosis and immunofluorescence staining revealed that early in apoptosis PKC delta accumulated in the nucleus. Addition of IFN-beta to T cells already deprived of survival factors or treated with anti-Fas antibody caused a rapid retranslocation of PKC-delta away from the nucleus. Furthermore, the generation of a constitutively active catalytic fragment by cleavage of PKC-delta by caspase 3 occurred only after translocation of full-length PKC-delta to the nucleus. IFN beta also inhibited caspase 3 and the proteolytic activation of PKC-delta. We conclude from these studies that nuclear translocation of PKC-delta is an early event in T cell apoptosis and that IFN-beta rapidly reverses this process. PMID- 10458776 TI - Recombinant human IgG molecules lacking Fcgamma receptor I binding and monocyte triggering activities. AB - Subclasses of human IgG have a range of activity levels with different effector systems but each triggers at least one mechanism of cell destruction. We are aiming to engineer non-destructive human IgG constant regions for therapeutic applications where depletion of cells bearing the target antigen is undesirable. The attributes required are a lack of killing via Fcgamma receptors (R) and complement but retention of neonatal FcR binding to maintain placental transport and the prolonged half-life of IgG. Eight variants of human IgG constant regions were made with anti-RhD and CD52 specificities. The mutations, in one or two key regions of the CH2 domain, were restricted to incorporation of motifs from other subclasses to minimize potential immunogenicity. IgG2 residues at positions 233 - 236, substituted into IgG1 and IgG4, reduced binding to FcgammaRI by 10(4)-fold and eliminated the human monocyte response to antibody-sensitized red blood cells, resulting in antibodies which blocked the functions of active antibodies. If glycine 236, which is deleted in IgG2, was restored to the IgG1 and IgG4 mutants, low levels of activity were observed. Introduction of the IgG4 residues at positions 327, 330 and 331 of IgG1 and IgG2 had no effect on FcgammaRI binding but caused a small decrease in monocyte triggering. PMID- 10458777 TI - Obligatory cross-talk with the tyrosine kinases assembled with the TCR/CD3 complex in CD4 signal transduction. AB - Dissection of the CD4 signal transduction pathway has revealed striking similarities with the TCR/CD3 pathway. Furthermore, downstream signaling by CD4 is impaired in cells lacking surface TCR, suggesting a role for the TCR/CD3 complex in CD4 signal transduction. We have investigated the molecular basis for the dependence of CD4 signaling on TCR/CD3 expression. Using the phosphotyrosine binding domains of the Shc adaptor and the Fyn kinase, which both participate in CD4 signaling, as baits, we show that CD4 induces tyrosine phosphorylation of a subset of the proteins phosphorylated in response to TCR/CD3 engagement. The phosphoprotein patterns were dramatically altered in cells defective for TCR/CD3 expression, and were recoverable by reconstitution of correctly assembled TCR, suggesting that CD4 uses TCR/CD3-associated tyrosine kinases to signal. Among the tyrosine kinases associated with the resting TCR/CD3 complex, only Fyn is activated following CD4 engagement. The failure of Fyn to become phosphorylated in cells defective for TCR expression underlines the unique role of TCR/CD3 associated Fyn in CD4 signal transduction. While no calcium mobilization was measurable in cells defective for TCR/CD3 expression in response to CD4 engagement, the Ras/MAP kinase pathway could be partially activated. Thus, CD4 activates at least two signaling pathways, and tyrosine kinases associated with the TCR/CD3 complex are key components of one of these pathways. PMID- 10458778 TI - Early cytokine induction by Plasmodium falciparum is not a classical endotoxin like process. AB - We have investigated the widely held view that malaria parasites induce pro inflammatory cytokines primarily through an endotoxin-like stimulatory effect on macrophages. We report that the pattern of cytokine production by non-immune human peripheral blood mononuclear cells following stimulation by Plasmodium falciparum-infected erythrocytes (Pfe) in vitro differs considerably from that induced by bacterial endotoxin. The Pfe-induced TNF response at day 1 is associated with a much higher level of IFN-gamma production and a much lower level of IL-12 p40 and IL-10 expression than a comparable endotoxin-induced TNF response. Both CD3(+) and CD14(+) populations are required for this early TNF response to Pfe, whereas the endotoxin-induced response is unaffected by depletion of the CD3(+) population. Pfe fails to stimulate the monocyte-like cell line MonoMac6 to express pro-inflammatory cytokines. These findings suggest that the early inflammatory response to malaria is critically dependent on lymphocyte subpopulations that play a lesser role in the response to bacterial endotoxin. PMID- 10458779 TI - Human mast cells produce matrix metalloproteinase 9. AB - Extracellular matrix-destructive enzymes, like matrix metalloproteinases (MMP), have been recognized in the process of inflammation and tissue remodeling and repair. The affected tissues often contain markedly increased numbers of mast cells. Although mast cells are capable of activating latent collagenase and proMMP, it has so far been unknown whether human mast cells themselves produce and secrete MMP9. In this study, MMP9 production by cord blood-derived cultured human mast cells and HMC-1 human mast cells was examined by reverse-transcriptase PCR, gelatin zymography and Western blot analysis using an antibody against MMP9. Cultured mast cells and HMC-1 cells treated with phorbol 12-myristate 13-acetate were shown to express MMP9 mRNA, and the cultured conditioned media from these cells showed gelatinolytic activity, identical with MMP9. Immunohistochemical examination was performed to detect MMP9 in tissue mast cells; mast cells localized in the skin, lung and synovial tissue showed strongly positive reactions for MMP9. Thus, these findings indicate that human mast cells can produce MMP9, which might contribute to extracellular matrix degradation and absorption in the process of allergic and nonallergic responses. PMID- 10458780 TI - Carbohydrate Mimetics: A New Strategy for Tackling the Problem of Carbohydrate Mediated Biological Recognition. AB - Useful strategies for the design of molecules to mimic carbohydrates have been developed over the past few years. Mimics of the target may contain new functional groups, a new scaffold, or both (in the schematic representation the natural ligand is shown on the left and the modified version on the right). Many examples of successful carbohydrate mimetics that interfere with sugar-protein and sugar-nucleic acid interactions are known. PMID- 10458781 TI - Tertiary Motifs in RNA Structure and Folding. AB - Specific tertiary structural motifs determine the complete architecture of RNA molecules (see picture for examples). Within the last few years a number of high resolution crystal structures of complex RNAs have led to new insights into the mechanisms by which these complex folds are attained. In this review the structures of these tertiary motifs and how they influence the folding pathway of biological RNAs are discussed, as well as new developments in modeling RNA structure based upon these findings. PMID- 10458782 TI - Synthesis of Diaryl Ethers: A Long-Standing Problem Has Been Solved. AB - A breakthrough in the synthesis of diaryl ethers has been achieved as shown in Equation (1). The coupling of phenols with aryl boronic acids in the presence of copper(II) acetate and a base proceeds under very mild conditions (room temperature) as described simultaneously by Evans et al. and Chan et al. Examples: R(1)=2-Cl, 2-I, 2-OMe, 4-tBu, 4-CH(2)CH(NHCOOtBu)CO(2)Me, 3,5-tBu(2); R(2)=2-Me, 3-OMe, 3-NO(2), 4-H, 4-F, 4-OMe. PMID- 10458783 TI - A Phospholipase with a Novel Catalytic Triad. AB - Catalytic triads are a long-standing paradigm of enzyme catalysis. By using a "matched mutation" approach, that is, a simultaneous exchange of the protein (through mutagenesis) and the substrate (through substitution of oxygen atoms by sulfur atoms) followed by enzyme kinetic analysis, a novel catalytic triad (see figure) with an unusual amino acid composition is now proposed for a phospholipase that fulfills a dual function in catalysis. PMID- 10458784 TI - Spontaneous Enantiomeric Resolution in a Fluid Smectic Phase of a Racemate. AB - Right- and left-handed homochiral domains segregate in a fluid smectic phase of the racemic (R*,S*)-beta-Me-TFMHPOBC analogue shown. This CF(3)-containing liquid crystalline compound exhibits an electrooptic response in a homogeneous cell, although no macroscopic dipole is expected to exist. Moreover, the homeotropic cells of this material exhibit striped domains, which are associated with fine stripes having two opposite tilt senses with respect to the predominant stripes. PMID- 10458785 TI - Towards Sixfold Functionalization of Buckminsterfullerene (C(60)) at Fully Addressable Octahedral Sites. AB - Selective C(60)-functionalizations that provide access to unusual multifunctional molecules are of interest in the construction of highly organized three dimensional assemblies. The temporary "masking" of three of the most reactive sites on C(60) by a bisdiene tether has allowed the facile and high-yielding formation of the fully differentiated trisadduct 1 and the interesting hexaadduct 2. PMID- 10458786 TI - Formation of an Effective Opening within the Fullerene Core of C(60) by an Unusual Reaction Sequence. AB - A large hole in a fullerene: The addition of dioxygen to the highly reactive 1,4 diaminobutadiene moiety of 1, formed from the reaction of C(60) with a rigid diazidobutadiene, results in the very efficient formation of an open fullerene (see the space-filling model) with the largest orifice created so far on a fullerene. The opening may be large enough to allow the smallest atoms, molecules, or ions to pass through. PMID- 10458787 TI - Self-Assembly of a Tetrahedral Lectin into Predesigned Diamondlike Protein Crystals. AB - Binding sites analogous to those of sp(3) carbon are presented by concanavalin A. This lectin has now been cross-linked with a bismannopyranoside which contains the C(2) spacer required to form the computer-modeled diamondlike three dimensional protein lattice shown in the picture. PMID- 10458788 TI - N-Substituted Corroles: A Novel Class of Chiral Ligands. AB - Although known for almost 35 years, N-substituted corroles have only now been recognized as being chiral. Several examples of these species were prepared in a facile two-step synthesis and separated into their enantiomers by HPLC. The zinc(II) complex of the corrole shown schematically was also synthesized. PMID- 10458789 TI - A Poly(para-phenylene) with Hydrophobic and Hydrophilic Dendrons: Prototype of an Amphiphilic Cylinder with the Potential to Segregate Lengthwise. AB - Langmuir monolayers are formed from an amphiphilically decorated poly(para phenylene), which indicates that its hydrophilic and hydrophobic parts segregate lengthwise along the polymer backbone in this nanometer-sized cylinder as illustrated in A. This polymer differs from known amphiphiles in that it consists of a linear, covalently bound sequence of "little" amphiphiles. It is much more rigid than common "polysoaps", which should increase its potential to aggregate, for example, into channels. PMID- 10458790 TI - The First Efficient Hydroaminomethylation with Ammonia: With Dual Metal Catalysts and Two-Phase Catalysis to Primary Amines. AB - Primary and unbranched secondary amines are obtained by the highly selective hydroaminomethylation of olefins with ammonia [Eq. (a)]. The selectivity is readily controlled with a new dual Rh/Ir catalyst in a two-phase system. PMID- 10458791 TI - A Novel Coordination Mode of 7-Methyl-7-sila-nido-undecaborate(1-). AB - B-N-M bridges, resulting from the activation of two B,H units, are present in the anion [Nb(MeSiB(10)H(8))(u-NMe(2))(2)Br(3)](-) (see structure depicted). Thus, this is an example for a hitherto unknown coordination mode in the chemistry of heteroborane clusters. PMID- 10458792 TI - Supramolecular Sensors for the Detection of Alcohols. AB - Cooperativity between different noncovalent host-guest interactions is the key for the successful generation of selective supramolecular sensors. A new class of cavitands capable of synergistic CH small middle dot small middle dot small middle dotpi interactions and hydrogen bonding has been designed and exploited for the detection of alcohol vapors using mass transducers (see schematic representation). PMID- 10458793 TI - The Valence States of Nickel, Tin, and Sulfur in the Ternary Chalcogenide Ni(3)Sn(2)S(2)-XPS, (61)Ni and (119)Sn Mossbauer Investigations, and Band Structure Calculations. AB - The hitherto controversial valence states of nickel and tin in the ternary chalcogenide Ni(3)Sn(2)S(2) (see structure) have been determined by photoelectron and Mossbauer spectroscopy ((61)Ni, (119)Sn). Results from band structure calculations confirmed that this shandite phase is a metal and that the approximate distribution of the valence electrons is (Ni(0))(3)(Sn(1)(II))(Sn(2)(II))(S(II-))(2). PMID- 10458794 TI - Samarium-Mediated beta-Elimination in Dihalo Alcohols: Diastereoselective Synthesis of (Z)-Vinyl Halides. AB - High diastereoselectivity in beta-elimination reactions of O-acetyl 1,1 dihaloalkan-2-ols to give (Z)-vinyl halides was achieved by using samarium diiodide [Eq. (1)]. The reaction was also highly diastereoselective and totally chemoselective when a mixture of diastereoisomers was used as substrate (X(1), X(2)=halogen; X(1) not equal X(2)). PMID- 10458796 TI - Functionalizable Alkylidenes: Tungsten Complexes of Phosphanyl-, Amino-, Alkynyl , and Tinalkylidenes and Their Dimetallic Derivatization. AB - The introduction of a heteroatom at the alkylidene carbon atom of calixarene complexes such as 1 results in an organometallic compound that shows properties of both Fischer carbenes and Schrock alkylidenes. Such modified alkylidenes allow access to dimetallic alkylidene complexes. For example, 1 reacts with [CuCOCl] to provide 2. PMID- 10458795 TI - Enantioselective Extraction of Dinitrophenyl Amino Acids Mediated by Lipophilic Deoxyguanosine Derivatives: Chiral Discrimination by Self-Assembly. AB - The transfer of potassium salts of dinitrophenyl amino acids from water to chloroform by the lipophilic guanosine derivative 1 takes place enantioselectively. Depending on the K(+):1 ratio, G-quartets of 1 self-assemble into octamers (O) or polymers. PMID- 10458797 TI - Diastereomeric Shape Recognition Using NMR Spectroscopy in a Chiral Liquid Crystalline Solvent. AB - Unambiguous assignment of the resonances of different diastereomers in a nonracemic mixture is possible with (1)H-decoupled (2)H NMR spectroscopy in a chiral liquid crystalline solvent. This is demonstrated with, for example, the alpha,alpha'-dideuterated diol shown in the picture. Even diastereomers with centers of chirality up to nine bonds apart can be discriminated. PMID- 10458798 TI - Photoregulation of the Formation and Dissociation of a DNA Duplex by Using the cis-trans Isomerization of Azobenzene. AB - The duplex-forming activity of an oligonucleotide has been photoregulated by making use of the isomerization of an azobenzene moiety in the side chain. When the azobenzene moiety is isomerized from the trans form to the cis form upon photoirradiation, the melting temperature of the duplex between the oligonucleotide and its complementary counterpart is significantly lowered, and the duplex is largely dissociated into two single-stranded oligonucleotides (shown schematically). PMID- 10458799 TI - Acylzirconocene Chloride as an "Unmasked" Acyl Anion: Enantioselective 1,2 Addition to alpha,beta-Unsaturated Ketone Derivatives. AB - The chiral monodentate phosphane ligand (R)-MOP facilitated the first enantioselective nucleophilic 1,2-addition of an "unmasked" acyl anion to a carbonyl group in the Pd(II)-catalyzed reaction of alpha,beta-unsaturated ketones with acylzirconocene chlorides [Eq. (1), 66 % ee, 88 % yield; (R)-MOP=(R)-2 (diphenylphosphanyl)-2'-methoxy-1,1'-binaphthyl]. PMID- 10458800 TI - Highly Enantio- and Diastereoselective Hetero-Diels-Alder Reactions Catalyzed by New Chiral Tridentate Chromium(III) Catalysts. AB - Even moderately nucleophilic dienes react with simple aldehydes in the presence of a new Cr(III) catalyst in a hetero-Diels-Alder reaction [Eq. (1)]. Tetrahydropyranyl products with up to three stereogenic centers are generated in near-perfect diastereoselectivities and with greater than 90 % ee (99 % ee for the example shown). TBAF=tetrabutylammonium fluoride; TBS=tert butyldimethylsilyl; TES=triethylsilyl. PMID- 10458801 TI - Copper(I) Complexes with a Proximal Aromatic Ring: Novel Copper-Indole Bonding. AB - A unique bonding of indole to a metal ion has been established. Two new tridentate N ligands with a pendent indole ring gave Cu(I) complexes 1 and 2, the latter of which exhibits the new form of bonding between the Cu(I) ion in a distorted tetrahedral geometry and the indole C(2)-C(3) moiety. The bond is dependent upon the length of the side chain and therefore the accessibility of the ring to the metal center. PMID- 10458802 TI - Self-Assembling Supramolecular Nanostructures from a C(60) Derivative: Nanorods and Vesicles. AB - Simple modification of solution conditions provides facile access to supramolecular fullerene nanostructures. The fullerene derivative shown self assembles to give nanorods or vesicles. The nanorods have diameters of 10-250 nm, depending on the counterion, and lengths greater than 70 um. If ultrasonication is used, no nanorods form, but vesicles result having diameters of 10-70 nm and wall thicknesses of 3-6 nm. PMID- 10458803 TI - Intra- and Intermolecular H/D Exchange in Aqueous Solution Catalyzed by Molybdocenes. AB - C-H bond activation in water catalyzed by water-soluble molybdocene complexes results in H/D exchange in alcohols [Eq. (1)]. Deuterium is incorporated intramolecularly in the Cp' methyl group of the metallocene as well as intermolecularly on the alpha-carbon atoms of the alcohols at moderate temperatures (Cp'=eta(5)-C(5)H(4)CH(3)). PMID- 10458804 TI - Ionization of Ozone/Chlorofluorocarbon Mixtures in Atmospheric Gases: Formation and Dissociation of AB - Complexes that undergo extensive molecular reorganization upon unimolecular metastable dissociation are obtained after ionization of atmospheric gases containing ozone and chlorofluorocarbons [Eq. (1)]. These unusual processes, which require fission of all bonds initially present in the CHX(2) unit (X=Cl, F) and combination of C with one of the O atoms of ozone, were demonstrated and their mechanism rationalized by the joint application of mass spectrometric and theoretical methods. PMID- 10458805 TI - The First General Method for Stille Cross-Couplings of Aryl Chlorides. AB - A "one-two punch" comprising two commercially available reagents, PtBu(3) and CsF, provides a practical and general solution for a long-standing limitation of the Stille reaction-the inability to couple inexpensive and readily available aryl chlorides [Eq. (1); R(1)=OMe, NH(2), o-Me, etc.; R=vinyl, allyl, Ph, Bu, etc.]. PMID- 10458806 TI - A Highly Active Catalyst for the Room-Temperature Amination and Suzuki Coupling of Aryl Chlorides. AB - A unique combination of steric and electronic properties appears to determine the effectiveness of phosphanyl-substituted biphenyls as ligands in palladium catalyzed aminations and Suzuki coupling of aryl chlorides at room temperature [Eq. (1)]. The oxidative addition step is greatly accelerated, and transmetalation (or Pd-N bond formation) and reductive elimination processes are facilitated. Use of these ligands allows for Suzuki coupling at very low catalyst loadings (as little as 10(-6) mol % Pd). R"=cyclohexyl, tert-butyl. PMID- 10458807 TI - Highly Active Ruthenium Catalysts for Olefin Metathesis: The Synergy of N Heterocyclic Carbenes and Coordinatively Labile Ligands. AB - Two N-heterocyclic carbene ligands at once may be one too many, at least if you intend to have highly active ruthenium catalysts for olefin metathesis. Density functional calculations recommend the replacement of the second carbene ligand in the successful ROMP catalysts 1 by coordinatively more labile ligands as in 2 or 3. In both cases, the catalytic activity is greatly improved. PMID- 10458808 TI - A One-Pot, One-Operation AB - Enantiomerically pure 2,1-benzothiazines are readily prepared by a one-pot, one operation procedure consisting of a palladium-catalyzed aryl amination of an ortho-bromobenzaldehyde with a sulfoximine followed by an intramolecular condensation reaction [Eq. (1)]. BINAP=1,1'-binaphthalene-2,2' diylbis(diphenylphosphane). PMID- 10458809 TI - Asymmetric Synthesis of Novel Ferrocenyl Ligands with Planar and Central Chirality. AB - A stereogenic center at the position beta to the metallocene backbone is present in ferrocenyl ligands 2, which are interesting for asymmetric catalysis. These planar-chiral compounds are accessible for the first time by a highly diastereoselective and enantioselective synthesis (de=93-97 %; ee>/=96 %) from the ferrocenyl ketones 1. A variety of donor groups (E(1)=Ph(2)P small middle dotBH(3), SMe, SiPr; E(2)=SMe, STol, SePh, Ph(2)P small middle dotBH(3), iPr(2)P small middle dotBH(3)) can be introduced as electrophiles. Tol=tolyl=CH(3)C(6)H(4). PMID- 10458810 TI - Total Synthesis of Quinoxapeptin A-C: Establishment of Absolute Stereochemistry. AB - The relative and absolute stereochemistry of the naturally occurring potent HIV reverse transcriptase (RT) inhibitors 1 and 2, quinoxapeptin A and B, were established by total synthesis. Their synthetic precursor 3 (dubbed quinoxapeptin C) was found to be a more potent HIV-1 RT inhibitor and to lack the potent cytotoxic activity characteristic of 1 and 2. PMID- 10458811 TI - Rhodium(I)-Catalyzed AB - The regiospecific introduction of substituents into the 4-position of 2,3 dihydroindoles (indolines), which is significant for the synthesis of various natural products and pharmaceuticals, was achieved by rhodium(I)-catalyzed cyclotrimerizations of 1 with acetylene to give 2. Up to four substituents can be introduced simultaneously into the indoline core by using this novel strategy. R=OH, OBzl, Otetrahydropyranyl, NHSO(2)C(6)H(4)CH(3); Ts=H(3)CC(6)H(4)SO(2). PMID- 10458812 TI - Tricyclopropylamine and Its Radical Cation. AB - A surprisingly high first vertical ionization energy (8.44 eV) and an unusually high oxidation potential are exibited by tricyclopropylamine (1)-in sharp contrast to triisopropylamine. These are attributed to the near-tetrahedral geometry of 1 at the N atom and the perpendicular orientation-with respect to the N-orbital axis-of the cyclopropyl groups. gamma-Irradiation of 1 led to the radical cation 1(.)(+), which, in accord with computations, has a planar C(3h) symmetrical structure. The EPR-spectroscopic and computational results disclose a dramatic, previously not reported, conformational change on going from 1 to 1(.)(+). PMID- 10458813 TI - Resolution of Racemic 1,2-Dibromohexafluoropropane through Halogen-Bonded Supramolecular Helices. AB - Halogen bonds, attractive intermolecular interactions between perfluoroalkyl bromides and bromide ions, are present in cocrystals of (-)-sparteinium hydrobromide (1) and (S)-1,2-dibromohexafluoropropane (2; shown schematically), and result in enantiopure and infinite supramolecular helices. The perfluorocarbon-hydrocarbon self-assembly allows the resolution of racemic 2. PMID- 10458814 TI - A Germanium Zeotype Containing Intratunnel Transition Metal Complexes. AB - A new zeolite-type structure is adopted by (NH(4))(+)[M(NH(3))(2)](+)(Ge(9)O(19))(2-) (M=Cu, Ag; shown in the picture). These compounds are the first microporous germanates containing a transition metal complex inside their tunnels. The large separation between the metal centers and the unhindered access of reactants to these active sites through uniformly sized channels make these materials a good point of departure for designing new catalysts. PMID- 10458815 TI - Donor/Acceptor Metallocenes: A New Structure Principle in Catalyst Design. AB - Suitable as highly effective polymerization catalysts is the new class of donor/acceptor metallocenes in which the rotation of the two pi ligands is restricted through the formation of a dative D(+)-->A(-) bond (see picture). Specifically optimized substitution patterns yield excellent properties for the synthesis of high-melting, highly crystalline thermoplastic materials, amorphous thermoplastic materials with high glass transition temperatures as well as polyolefin elastomers with low glass T(g). PMID- 10458816 TI - Macrolide Analogues of the Novel Immunosuppressant Sanglifehrin: New Application of the Ring-Closing Metathesis Reaction. AB - Macrocycles containing a conjugated 1,3-diene moiety have been synthesized for the first time in good yields by the ring-closing metathesis reaction [Eq. (1)]. The new compounds represent cyclophilin-binding, simplified analogues of the macrocyclic core of sanglifehrin A, an immunosuppressant which binds with high affinity to cyclophilin. PMID- 10458817 TI - Total Synthesis of Sanglifehrin A. AB - The immunosuppressive agent sanglifehrin A has been prepared for the first time by total synthesis. The construction of the macrocyclic unit of the target molecule was achieved through a selective intramolecular Stille coupling, and the spirolactam unit by Paterson-aldol reactions. The final steps involve an intermolecular Stille coupling and the opening of the internal acetal unit. This convergent synthesis opens the way for the synthesis of libraries of novel sanglifehrin analogues for biological screening. PMID- 10458818 TI - Frequency of unacceptable results in point-of-care testing. PMID- 10458819 TI - A portrait in history. The anatomical legacy of Dr Sommerring. PMID- 10458820 TI - Evaluation of a fully automated high-performance liquid chromatography assay for hemoglobin A1c. AB - INTRODUCTION: Measurement of hemoglobin A1c (HbA1c) is used as an objective measure of long-term blood glucose control in diabetic patients. Recent improvements in automation combined with new recommendations for precision and accuracy have caused us to reevaluate our methods for measuring HbA1c. OBJECTIVE: We evaluated a newly automated high-performance liquid chromatography (HPLC) instrument for measurement of HbA1c (Tosoh A1c 2.2 Plus Glycohemoglobin Analyzer, Tosoh Medics, Foster City, Calif) and compared the results obtained by HPLC to those obtained with an immunoassay (Hitachi 911, Boehringer Mannheim Corporation, Indianapolis, Ind). RESULTS: The Tosoh analyzer was found to be linear in a range of 5.3% to 17% and had a throughput of 20 samples per hour. HbA1c results for 102 patient samples by the 2 techniques showed good correlation, with a slope of 0.87 and an intercept at 1.27% +/- 0.15%. Both the total and within-run coefficients of variation were consistently lower for the HPLC method compared with the immunoassay method. The HPLC method produces a chromatogram that shows the different hemoglobin fractions, allowing identification of abnormal hemoglobin variants. In heterozygous individuals, HbA1c measurements are made with no interference from the hemoglobin variant. In the case of homozygous or doubly heterozygous hemoglobin variants, the Tosoh HPLC identifies the hemoglobin variants as such and correctly does not report a HbA1c value in the presence of a markedly decreased amount of hemoglobin A. CONCLUSIONS: The Tosoh HPLC provides adequate throughput and improved precision, and the method is traceable to the Diabetes Control and Complications Trial. PMID- 10458821 TI - The use of monoclonal antibody R92F6 and polymerase chain reaction to confirm the presence of parvovirus B19 in bone marrow specimens of patients with acquired immunodeficiency syndrome. AB - BACKGROUND: Parvovirus B19 infection is a cause of chronic anemia and red cell aplasia in patients with acquired immunodeficiency syndrome (AIDS) and in other immunocompromised hosts. Anemia in AIDS patients has a multifactorial etiology, with parvovirus B19 infection being an infrequent but nevertheless treatable cause. Therapy with intravenous immune globulin can result in rapid improvement of parvovirus-induced anemia. This treatment is expensive, therefore accurate and rapid confirmation of parvovirus infection is important in providing appropriate and cost-effective therapy. METHODS: Bone marrow samples from 2 AIDS patients with severe anemia and reticulocytopenia were studied. Bone marrow morphology and serologic studies were evaluated for parvovirus B19 infection. An immunohistochemical method using a monoclonal antibody, R92F6, to B19 capsid proteins was utilized on decalcified, B5-fixed, paraffin-embedded bone marrow biopsies. Bone marrow aspirate cells were examined by electron microscopy for evidence of viral particles. In addition, polymerase chain reaction (PCR) studies using a nested PCR assay to the parvovirus B19 viral genome were performed in a case for which fresh cells were available. RESULTS: Bone marrow findings included marked erythroid hypoplasia with characteristic giant pronormoblasts and intranuclear inclusions. Serologic studies were negative in one case, while the second case showed positive parvovirus B19 immunoglobulin M antibody. Immunohistochemical studies for parvovirus B19 were positive in both cases. The presence of intranuclear virions was demonstrated by electron microscopy and was confirmed by PCR analysis. Both patients were treated with intravenous immune globulin, and subsequent improvement was noted. CONCLUSIONS: Both immunohistochemistry and PCR studies on bone marrow specimens from AIDS patients with anemia are rapid and sensitive methods for the confirmation of parvovirus B19 infection. They are valuable tools, particularly when serologic studies are negative. When PCR is not available, immunohistochemical methods can be useful. The rapid confirmation of parvovirus B19 infection will allow for early and cost effective therapy. PMID- 10458822 TI - Intrathecal methotrexate-induced megaloblastic anemia in patients with acute leukemia. AB - OBJECTIVE: To evaluate the occurrence of megaloblastic anemia induced by the infusion of therapeutic or prophylactic methotrexate in patients with acute leukemia. DESIGN: Data on 3 patients with acute leukemia receiving intrathecal methotrexate were prospectively analyzed. SETTING: Large tertiary-care center. RESULTS: All 3 patients with acute leukemia developed megaloblastic anemia confirmed by examination of the bone marrow aspirate and biopsy. Two of the 3 patients had low folic acid levels, while all patients had normal serum B(12) levels. All patients responded favorably to a therapeutic trial of folic acid. The median time for recovery of the hematologic parameters in these patients was 7 days. CONCLUSIONS: Intrathecally administered methotrexate may result in megaloblastic changes in the bone marrow of leukemic patients. The morphologic clues suggestive of folate deficiency in patients with acute leukemia may be masked by coexisting factors, such as the effects of cytotoxic treatment, prior transfusions, or persistent changes from the leukemic clone itself. Caution should be exercised to avoid attributing these changes to the neoplastic process, since the prognosis and treatment for the conditions involved are totally different. Repeat examination of the bone marrow, obtaining folic acid and vitamin B(12) levels, and a therapeutic trial of folic acid may help identify and reverse these changes. PMID- 10458823 TI - Endoscopic biopsy pathology of Helicobacter pylori gastritis. Comparison of bacterial detection by immunohistochemistry and Genta stain. AB - OBJECTIVES: To describe the endoscopic biopsy pathology of Helicobacter pylori gastritis, compare bacterial detection by immunohistochemistry using a specific antibody with the Genta stain, and to compare the relative costs of the 2 techniques. DESIGN: One hundred cases of gastritis identified as positive for H pylori by Genta stain and 100 cases considered negative by the same technique were stained using an anti-H pylori-specific polyclonal antibody. Laboratory reagent and labor costs for the 2 methods were compared. RESULTS: Chronic active gastritis with lymphoid follicles was significantly associated with H pylori infection (P <.0001). The immunohistochemical method had a sensitivity of 97% and a specificity of 98% compared with the Genta stain, with strong agreement for grading density of organisms (kappa = 0.85; P <.001). Reagent costs were similar for both methods, but immunohistochemistry using an autoimmunostainer required less dedicated technical time and hence was less expensive than the Genta stain. CONCLUSIONS: Immunohistochemistry using a specific antibody is an accurate and cost-effective method for H pylori detection in gastric biopsies. PMID- 10458824 TI - Epithelioid and spindle-celled leiomyosarcoma of the heart. Report of 2 cases and review of the literature. AB - BACKGROUND: Primary cardiac leiomyosarcomas are rare. Isolated reported cases and small series generally describe spindle-celled, high-grade tumors with poor short term survival; however, the pathologic features of many of these tumors are incompletely documented. The authors report in detail the clinicopathologic features of 2 relatively low-grade epithelioid and spindle-celled primary cardiac leiomyosarcomas. METHODS: Cases 1 and 2 were studied using standard histochemical and immunohistochemical techniques, and case 1 was examined by electron microscopy. The literature was reviewed with regard to primary cardiac leiomyosarcomas. RESULTS: Both tumors showed epithelioid and spindle-celled areas. The tumor in case 1 was low grade, and the tumor in case 2 was predominately low grade with a high-grade focus. A review of 28 reported cases revealed a wide age range (mean, 43 years), equal male-to-female ratio, and a predilection for the left atrium (48%). Follow-up of reported cases with fewer than 5 mitoses per 10 high-power fields showed a mean survival of 22 months compared with a 9-month mean survival for all others. CONCLUSIONS: Short-term follow-up of reported cases of high-grade cardiac leiomyosarcoma suggests a poor prognosis. Long-term follow-up in our case 2, along with follow-up of reported cases that were histologically similar to our cases, suggests that cardiac leiomyosarcomas with low-grade features or mixed low- and high-grade features also have a poor overall long-term survival, with a high rate of local recurrence and systemic spread. PMID- 10458825 TI - Mucocutaneous angiomyolipoma. A report of 2 cases arising in the nasal cavity. AB - OBJECTIVE: Angiomyolipoma (AML) is a distinctive tumor that usually occurs in the kidney and rarely in the liver. Angiomyolipoma of other sites is extremely rare, and, to our knowledge, only 2 cases have been described in the nasal cavity. We present 2 additional cases of AML of the nasal cavity and discuss the difference between them and renal AML. CASE REPORTS AND PATHOLOGIC FINDINGS: Two tumors in a 66-year-old man and an 88-year-old woman without tuberous sclerosis are described. They showed 20-mm, well-circumscribed, polypoid shapes. Histologically, they were composed of mature smooth muscle cells, fat cells, and various-sized blood vessels. In addition, aggregated small lymphocytes were noted. Neither epithelioid smooth muscle cells nor HMB45 immunoreactivity was seen in either case. CONCLUSIONS: The clinicopathologic features of AML of the nasal cavity are distinct from renal and hepatic AML and are common to those of AML arising in the skin and oral and pharyngeal mucosa. The term mucocutaneous angiomyolipoma is thought to be appropriate to express these characteristic tumors. PMID- 10458826 TI - Histopathologic features and MIB-1 labeling indices in recurrent and nonrecurrent meningiomas. AB - BACKGROUND: Predicting the behavior of meningiomas based on histopathologic features alone has remained problematic. DESIGN: This study retrospectively compared several histopathologic features and MIB-1 labeling indices (LIs) in recurrent meningiomas with those of nonrecurrent meningiomas. Six histopathologic features, including mitoses, necrosis, loss of architectural pattern, hypervascularity/hemosiderin deposition, prominent nucleoli, and nuclear pleomorphism, were compared between 32 recurrent and 27 nonrecurrent meningiomas using Fisher exact tests. MIB-1 LIs (% positive tumor cell nuclei) were compared using the Wilcoxon rank sum test. RESULTS: The patients in the recurrent group included 26 women (mean age, 55 years), who developed 1 to 5 recurrences. Time intervals to the first recurrence ranged from 5 to 183 months (mean, 55 months). The nonrecurrent group included 21 women (mean age, 56 years), with follow-up ranging from 88 to 124 months (mean, 109 months). Of the histopathologic features evaluated, statistically significant differences between the recurrent and nonrecurrent groups were found only with respect to prominent nucleoli (P =.024) and nuclear pleomorphism (P <.001), both of which were more common in the recurrent group. In the recurrent group, 9 tumors were considered malignant (defined by brain invasion or metastasis) versus 2 of the nonrecurrent meningiomas. Nineteen percent of nonrecurrent tumors versus 41% of recurrent tumors had 2 or more of the 6 histopathologic features. MIB-1 LIs in the nonrecurrent group ranged from 0 to 8.3 (mean, 1.5) and were generally lower than those in the recurrent group (range, 0-32.5; mean, 5.4); no statistical difference was identified between these groups. No statistically significant difference with regard to histology or MIB-1 LIs was noted between the initially excised recurrent tumor and the most recently resected recurrence. CONCLUSIONS: Of the histopathologic features examined, only prominent nucleoli and nuclear pleomorphism were found to be statistically more common in recurrent than nonrecurrent meningiomas. The mean MIB-1 LI was higher in the recurrent than in the nonrecurrent group, although there was no statistical difference between means and there was clear overlap with regard to MIB-1 LI ranges. PMID- 10458827 TI - The myoepithelial immunophenotype in 135 benign and malignant salivary gland tumors other than pleomorphic adenoma. AB - BACKGROUND: We have previously studied the immunoreactivity of 3 novel smooth muscle-specific proteins, alpha-smooth muscle actin, smooth muscle myosin heavy chains, and calponin, to assess myoepithelial differentiation in pleomorphic adenomas. OBJECTIVE: To further expand our knowledge of myoepithelial differentiation in other benign and malignant salivary gland tumors. DESIGN: Formalin-fixed paraffin sections of 135 salivary gland tumors with associated normal glands were stained with monoclonal antibodies using the avidin-biotin complex immunoperoxidase method and enzymatic and microwave heat-induced epitope retrieval. RESULTS: In adenoid cystic carcinomas and epithelial-myoepithelial carcinomas, all 3 markers exclusively highlighted the myoepithelial cell components and the epithelial cells were entirely negative. No immunostaining was detected in canalicular adenomas, oncocytomas, Warthin tumors, acinic cell carcinomas, mucoepidermoid carcinomas, squamous cell carcinomas, and polymorphous low-grade adenocarcinomas. Salivary duct carcinomas and adenocarcinomas, not otherwise specified had a distinctive pattern of uniform periductal staining of reactive myofibroblastic cells, and in salivary duct carcinomas some ducts retained a peripheral immunoreactive myoepithelial cell layer. CONCLUSION: Immunoreactivity for these 3 smooth muscle-specific proteins confirms the known neoplastic myoepithelial component of adenoid cystic carcinomas and epithelial myoepithelial carcinomas. The consistently positive staining pattern in adenoid cystic carcinomas may be diagnostically useful in discriminating histologically similar but consistently negative polymorphous low-grade adenocarcinomas. Periductal linear staining in adenocarcinoma, not otherwise specified and salivary duct carcinomas is distinctive and appears to represent a tight cuff of myofibroblasts associated with the infiltrating glands. PMID- 10458829 TI - Immunoexpression of villin in neuroendocrine tumors and its diagnostic implications. AB - BACKGROUND: Villin, a 95-kd cytoskeletal protein associated with axial microfilament bundles of brush border microvilli, is mostly restricted to intestinal glandular tumors. Villin immunoexpression was recently observed in a small number of carcinoids of the intestinal tract and lung, but its significance in a broad category of neuroendocrine tumors has not been evaluated. DESIGN: A total of 114 neuroendocrine tumors of different origins were tested for villin expression. They included gastrointestinal carcinoids (n = 30), lung carcinoids (n = 15), small cell carcinomas of the lung (n = 24), small cell carcinomas of other sites (n = 15), islet cell tumors (n = 8), Merkel cell carcinomas (n = 6), paragangliomas (n = 6), and others (n = 10). Nine round cell sarcomas were tested as well. RESULTS: Villin immunoreactivity was present in 85% of gastrointestinal carcinoids and small cell carcinomas, but was found in only 40% of lung carcinoids. Other tumors tested were virtually negative for villin. In general, while cytoplasmic reactivity was most common, a characteristic apical membranous pattern simulating brush border was seen in 76% of the gastrointestinal carcinoids and in 50% of the lung carcinoids. CONCLUSIONS: We found that villin was predominantly restricted to gastrointestinal neuroendocrine tumors (excluding islet cell tumors), although a small number of bronchial carcinoids may be positive as well. These results suggest a role for villin in the differential diagnosis of neuroendocrine tumors. PMID- 10458828 TI - BK virus renal infection in a patient with the acquired immunodeficiency syndrome. AB - BACKGROUND: We describe herein a patient with the acquired immunodeficiency syndrome and renal failure due to biopsy-proven BK virus (BKV) infection. Three months after the diagnosis of the renal viral infection, his condition remained unchanged. Although BKV has previously been shown to be associated with ureteral stenosis and renal damage in renal transplant patients, to our knowledge, the literature contains only 3 cases describing the presence of BKV lesions in the kidneys of immunosuppressed patients who had not undergone transplantation. METHODS: The presence of BKV infection was demonstrated by means of histology, immunohistochemistry with polyclonal anti-SV40 antibody, immunoelectron microscopy, polymerase chain reaction, and enzymatic cleavage with BamHI. RESULTS: Histologic examination revealed interstitial inflammatory infiltrates and tubules with enlarged and eosinophilic nuclei. CONCLUSIONS: The high frequency of latent BKV infection and its reactivation during immunosuppression suggest that the possibility of its involvement in renal damage should be considered in immunocompromised patients. PMID- 10458830 TI - Performance of a fluid-based, thin-layer papanicolaou smear method in the clinical setting of an independent laboratory and an outpatient screening population in New England. AB - BACKGROUND: A patented, fluid-based, thin-layer method for preparation of Papanicolaou (Pap) smears (ThinPrep Pap test) has been reported to be significantly more effective than the conventional smear invented by George Papanicolaou. We tested this position by comparing the cytologic diagnosis and specimen adequacy results obtained using the ThinPrep method with data from conventional Pap smears obtained from a similar population. METHODS: Test results of 56 339 ThinPrep specimens were compared with results from 74 756 conventional smear cases obtained from the same sources in a corresponding period of the previous year. RESULTS: The use of ThinPrep for cervicovaginal cytology produced a 75.14% increase in the detection of low-grade squamous intraepithelial lesions and higher diagnoses. Detection of low-grade squamous intraepithelial lesions increased by 71.65% (from 1.58% to 2.71%), and detection of high-grade squamous intraepithelial lesions increased by 102.54% (from 0.26% to 0.52%). There was a 39.11% decrease in the atypical squamous cells of undetermined significance intraepithelial lesion ratio (from 2.07 to 1.26). There were also marked decreases in the number of specimens categorized as "satisfactory but limited," owing to obscuring inflammation (-94.34%), obscuring blood (-99.84%), and poor fixation (-99.25%). CONCLUSION: ThinPrep produced increased detection of premalignant precursors while improving specimen adequacy. PMID- 10458831 TI - Free drug measurements. When and why? An overview. PMID- 10458832 TI - Continuous quality improvement for point-of-care testing using background monitoring of duplicate specimens. AB - CONTEXT: Despite compliance with quality control standards, concerns remain as to the accuracy and reliability of point-of-care testing. OBJECTIVE: To assess a practical method for quality improvement using the context in which point-of-care testing is done. DESIGN: Quality measures for point-of-care testing, making use of natural duplication of results obtained by other testing methods, were used to monitor testing quality and evaluate quality improvement interventions. SETTING: Five adult intensive care units (total of 88 beds) in a large academic medical center, using point-of-care testing for blood gases, electrolytes, and hematocrit levels. PARTICIPANTS: Nurses performing bedside testing and laboratory personnel assigned the responsibility for supervising their performance. INTERVENTIONS: Quality of testing was monitored continuously, and, where problems were identified, training and support interventions implemented, and their effects evaluated. MAIN OUTCOME MEASURES: Improvement in correlation coefficients and regression parameters of point-of-care hematocrit and potassium testing results compared with contemporaneous results from the core laboratory. RESULTS: The initial survey found point-of-care potassium levels were tightly correlated with core laboratory results (r = 0.958). Baseline correlation coefficients and regression parameters for point-of-care hematocrit levels compared with core laboratory values varied widely from unit to unit. The intensive care units with the highest variances of bedside vs core laboratory testing received targeted interventions. Follow-up yielded evidence of dramatic improvement; 1 unit experienced an increase in correlation from 0.50 to 0.95. CONCLUSIONS: The findings suggest that, when point-of-care testing is highly dependent on operator technique, targeted interventions can resolve problems and provide reliable results at the bedside. PMID- 10458833 TI - Myiasis secondary to Sermatobia hominis (human botfly) presenting as a long standing breast mass. AB - A case of a 54-year-old woman who presented with a breast mass is reported. Histologically, a chronic granulomatous inflammatory response was observed. The response was associated with an organism diagnosed as a fly larva, Dermatobia hominis (human botfly). The incidence of myiasis, infestation by fly larvae, presenting as a long-standing breast mass and mimicking a neoplasm is extremely rare, especially in the United States. PMID- 10458834 TI - Osteosarcoma arising in a solitary osteochondroma of the fibula. AB - We present a case of osteosarcoma arising in an osteochondroma of the right fibula in a 30-year-old woman. The available radiographic studies of the lesion were not suggestive of malignant transformation. The lesion and underlying bone were excised. Histologic examination showed a conventional high-grade osteoblastic osteosarcoma that focally eroded the fibrocartilaginous cap. The patient received postoperative chemotherapy and shows no evidence of disease 27 months following operation. The occurrence of osteosarcoma in an osteochondroma is an extremely rare event, and only a few cases are on record in the literature. PMID- 10458835 TI - Coinfection of visceral leishmaniasis and Mycobacterium in a patient with acquired immunodeficiency syndrome. AB - We report a case of coinfection of visceral leishmaniasis and Mycobacterium avium intracellulare in the same lesions in the small bowel and bone marrow of a 33 year-old man with acquired immunodeficiency syndrome who complained of abdominal pain and chronic diarrhea. The duodenal mucosa and bone marrow biopsy specimens showed numerous foamy macrophages packed with two forms of microorganisms that were identified histologically and ultrastructurally as Leishmania and Mycobacterium species. Visceral leishmaniasis is rarely suspected in patients residing in nonendemic countries including the United States. It should be included in the differential diagnosis for opportunistic infection in patients with acquired immunodeficiency syndrome. An appropriate travel history is important. To our knowledge, this is the first reported case showing coinfection of visceral leishmaniasis and Mycobacterium avium-intracelluulare in the same lesion in a patient with acquired immunodeficiency syndrome. PMID- 10458836 TI - Papillary thyroid carcinoma with nodular fasciitis-like stroma. Pitfalls in fine needle aspiration cytology. AB - Papillary thyroid carcinoma with nodular fasciitis-like stroma is one of the rare variants of papillary thyroid carcinoma. The problems posed by the exuberant nodular fasciitis-like stroma, which obscures the neoplastic nature of the tumor, are recognized in surgical pathology but have received little attention in the cytopathology literature. We report a rare case of papillary thyroid carcinoma in which nodular fasciitis-like stroma posed difficulty on fine-needle aspiration cytology. The differential diagnosis of fibroproliferative processes in thyroid fine-needle aspirations is also discussed. PMID- 10458837 TI - Congenital gastrointestinal pacemaker cell tumor. AB - The interstitial cells of Cajal complex within the gut wall function as a pacemaker to direct peristalsis. Their neoplastic counterpart is the gastrointestinal pacemaker cell tumor, a spindle and/or epithelioid cell mesenchymal tumor previously known as gastrointestinal stromal tumor or incorrectly called leiomyosarcoma in some cases of older reports. Although numerous cases of gastrointestinal leiomyosarcomas have been documented in the English-language literature, no pediatric case of gastrointestinal stromal tumor or gastrointestinal pacemaker cell tumor has, to our knowledge, been recorded. Herein, we report a case of congenital gastrointestinal pacemaker cell tumor confirmed by immunohistochemistry and electron microscopy in a full-term male newborn. PMID- 10458838 TI - Epithelioid hemangioendothelioma, multiple focal nodular hyperplasias, and cavernous hemangiomas of the liver. AB - Malignant vascular neoplasms of the liver are uncommon. We report the case of a young woman who developed an epithelioid hemangioendothelioma of the liver associated with multiple focal nodular hyperplasias and hepatic cavernous hemangiomas. Such an unusual association is probably not fortuitous and could support the theory that focal nodular hyperplasia is a reaction to an abnormal vascular supply rather than a true neoplasm. PMID- 10458839 TI - Benign fibroepithelial polyps of the renal pelvis. AB - Benign fibroepithelial polyps of the renal pelvis are rare. Although many ureteral fibroepithelial polyps have been reported, there are, to our knowledge, only 20+ reported cases in the renal pelvis. We report herein the clinical and pathologic features of fibroepithelial polyp of the renal pelvis and provide a detailed review of the literature. PMID- 10458840 TI - Pathologic quiz case. Extrarenal rhabdoid tumor. PMID- 10458841 TI - Images in pathology. Benign meningioma. PMID- 10458842 TI - Images in pathology. Placental transmogrification of the lung. PMID- 10458844 TI - I left my shoes in San Francisco. PMID- 10458845 TI - Serving on a board overseeing a closing institution. AB - Swerving on a board when an institution closes provides one with a wealth of frustrations and anxieties, both concerns for the institution, its employees, its clients, and oneself as possibly financially vulnerable. In today's economy more and more board members may find themselves faced with this uncomfortable task. Hopefully this article will make such a board member aware of some of the pitfalls and some of the necessary actions, minimizing the damage and chaos that inevitably results. Even when a closing is well orchestrated, the process is painful, but a well-designed board oversight plan can minimize problems and frustrations. PMID- 10458846 TI - Tracing our rituals in nursing. PMID- 10458847 TI - The advanced nurse practitioner. Struggling toward a conceptual framework. AB - For our committee, the exercise of exploring the role of the nurse practitioner (as well as the roles of our other advanced practice nurses) was a welcome intellectual task. It confirmed the commitment to our programs as well as giving us an intellectual base on which we could agree, creating a unity among the diversity in our many programs, each operating on what we term (using the language first applied in nursing by Eleanor Barba, RN, EdD) "boutique design," each operating rather independently in spite of some shared courses and modules. It was rewarding to find a commonality beneath our differences. We look forward to further revision of our work by the entire faculty. PMID- 10458848 TI - A feminist analysis of the theories of etiology of depression in women. AB - Women in the United States are more likely to be hospitalized for depression than be diagnosed with breast cancer during their lifetime, despite the continued underrecognition and underdiagnosis of depression. According to the DSM-IV, unipolar depression is a severe mood disorder characterized by a loss of pleasure in most activities, along with one or a combination of associated symptoms. Women between the ages of 20 and 45 are most prone to depression, with the incidence declining as they age. The likelihood of a correct diagnosis and the method of treatment of depression, as is true for other conditions and illnesses, is very much dependent upon the belief in its etiology; therefore, the theories of etiology warrant our serious consideration. Depression in women is explained in the literature most recently by biochemical explanations, though Freud's classic "female masochism" theory is still accepted in updated forms. Conflicting social roles and expectations, continued violence against women and children, and extreme disparities in socioeconomic opportunities and conditions between men and women are also cited as reasons for women's very high rates of depression in the U.S. Primary prevention through assessment for the predictors of depression offers the best hope for the promotion of mental health in women. During the course of routine health care interactions, nurses in primary care settings are potentially in the best position to assess each woman encountered for depression risk factors or symptoms. A few important questions asked by the nurse in the course of taking a brief health history or vital signs--such as where and with whom the woman lives; where she works (inside and/or outside the home) and if she finds the work fulfilling; how she supports herself financially; when, how much, and how well she sleeps; and what kinds of emotional support networks she has- may reveal enough to warrant a more detailed and specific assessment or a referral for treatment. Psychoeducation, often administered by psychiatric RNs, includes consciousness raising and self-help groups, women's studies courses, and the establishment of support networks. These, as well as various forms of feminist-based interpersonal psychotherapy, are oft-cited approaches to aiding at risk women clients. Given the willingness of many women clients to discuss their mental health status with a caring and interested nurse, perhaps the most positive intervention nurses can make is to listen, be supportive, and take seriously the pressures on women to be at once everything to everyone and no one at all. PMID- 10458849 TI - Role of middle range theory for nursing research and practice. Part I. Nursing research. PMID- 10458850 TI - The cost control heroism. Strategies to reduce operating expenses. PMID- 10458851 TI - Interview with Harriet R. Feldman, PhD, RN, FAAN. Interview by Barbara Stevens Barnum. PMID- 10458852 TI - Interview with Joyce J. Fitzpatrick, PhD, RN, FAAN. Interview by Barbara Stevens Barnum. PMID- 10458853 TI - Unmanaged care. PMID- 10458854 TI - Ethical leadership in managed care. Creating a new vision. PMID- 10458855 TI - Interview with Carolyn S. Zagury, MS, RN, CPC. Nurse entrepreneur and publisher. Interview by Barbara S. Barnum. PMID- 10458856 TI - The role of middle-range theory for nursing research and practice. Part 2. Nursing practice. PMID- 10458857 TI - Changing expectations towards nursing, family, and life chances. A cohort analysis of nurses in the 20th century. PMID- 10458858 TI - Cooperation in community health nursing. There is more to cooperation than doing things together. PMID- 10458859 TI - Partnership--a value at twice the price. PMID- 10458860 TI - Acute care nurse practitioners. An idea whose time has come? Point. PMID- 10458861 TI - ACNPs. An idea whose time has come? Counterpoint. PMID- 10458862 TI - Consulting and information technology. PMID- 10458863 TI - Primer for philanthropy. PMID- 10458864 TI - Interview with Dr. Ursula Springer. Interview by Barbara Stevens Barnum. PMID- 10458865 TI - Mergers. Your role as a nurse executive and leader of the system. PMID- 10458866 TI - Managing conflict. Is there a full moon? PMID- 10458867 TI - Case management. A system for improving outcomes. Point. PMID- 10458868 TI - Case management. Is it improving health outcomes? Counterpoint. PMID- 10458869 TI - Improving patient satisfaction through focus group interviews. AB - Long used by the social science community, focus groups have been increasingly used by nursing leaders to obtain information and feedback. Use of the focus group interview (FGI), a qualitative research method, allows the nurse executive the opportunity to obtain data on a variety of topics. In order for the findings to be meaningful, however, the researcher must adhere to basic FGI design and data analysis. Like other research methodologies, the focus group offers advantages and disadvantages that must be evaluated for appropriateness and feasibility. The article illustrates the use of the FGI to spark the creation of patient-focused care and significant increases in patient satisfaction in an academic medical center. PMID- 10458870 TI - Models and strategies of collaboration across countries in doctoral education. AB - This paper addresses the globalization of nursing, especially as it pertains to doctoral nursing education. Its purpose is to consider the issues surrounding cross-country doctoral education as identified by doctoral students and to suggest models and strategies that could be used to facilitate cross-country doctoral education. This paper is based on published sources, conversations with doctoral students and personal observations. Nursing, in order to become globally relevant, should develop an independent body committed to cross-country doctoral education. This body should be committed to facilitating and organizing programs that provide opportunities for students and faculty to attain experiences abroad. Several models incorporated under a comprehensive strategy are suggested in this paper. PMID- 10458871 TI - Interview with Diane O. McGivern. Vice Chancellor New York State Board of Regents. Interview by Barbara Stevens Barnum. PMID- 10458872 TI - The frontiers of global nursing. Need for knowledge development. PMID- 10458873 TI - Why aren't more nurses leaders? A consumer's perspective. AB - Nurse leaders are made, not born. It seems to this patient (and we all are likely to be patients at some time) that not enough nurses are leaders. I don't know if I am right, but if I am, then what it takes to be a nurse leader is for a good number of you to study the need, find specific issues that call for leadership action, and take the lead in effecting the change. Nurses have always had the power, but not enough of your have been organized and have learned how to use it. My experience as a patient has led to this attempt to look at some of the issues surrounding nursing leadership. I hope that this perspective is a stimulus to change for the betterment of patient care. PMID- 10458874 TI - The evolution of exaggerated sexual swellings in primates and the graded-signal hypothesis. AB - Females of some Old World primate taxa advertise their sexual receptivity with exaggerated sexual swellings. Although a number of hypotheses have been proposed, the function of this conspicuous trait remains unsolved. This review updates information on the phylogenetic distribution of exaggerated swellings and identifies aspects of the morphology, physiology and behaviour of species with this conspicuous trait. Some of these patterns represent new information, while other patterns have been previously identified, but not in ways that control for phylogeny. This review shows that exaggerated swellings are correlated with some features that serve to confuse paternity certainty among males, while other features tend to bias paternity towards more dominant males. Hypotheses for the evolution of exaggerated swellings are then reviewed and critically evaluated. Individually, no single hypothesis can account for all the patterns associated with exaggerated swellings; however, a combination of different hypotheses may explain the contradiction between confusing and biasing paternity. I suggest that exaggerated swellings can be viewed as distributions representing the probability of ovulation (the graded-signal hypothesis). In the context of this probabilistic model, exaggerated swellings enable females to manipulate male behaviour by altering the costs and benefits of mate guarding, so that dominant males tend to guard only at peak swelling, but females can mate with multiple males outside peak swelling to confuse paternity. This hypothesis makes testable predictions for future comparative and observational research. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10458875 TI - Postcopulatory sexual selection in Mediterranean fruit flies: advantages for large and protein-fed males. AB - Previous laboratory studies of Mediterranean fruit flies, Ceratitis capitata (medflies), have identified large size and protein feeding as positive influences on the ability of males to secure copulations. In this study, we investigated whether large and protein-fed males experience additional advantages in terms of amount and distribution of sperm stored by mates. We also examined relationships between copula duration and sperm storage. Mates of large and protein-fed males were more likely to store sperm and to store more sperm than mates of small and protein-deprived males. Probability of sperm storage was associated with copula duration; all copulations lasting less than 100 min failed whereas 98% lasting longer than 100 min succeeded. Copulations involving sperm storage were longer if males were small or protein deprived or if the female was large, although there was no evidence of a relationship between copula duration and total sperm storage. Evidence from related studies suggests that variation in latency until sperm transfer, caused by size and diet, is a likely explanation for varying copula duration. Sperm tended to be stored asymmetrically between the female's two spermathecae, consistent with a mating system in which females maintain isolated populations of sperm from different males and later select between them. Storage was less asymmetric when large numbers of sperm were stored but there was little evidence that male size or diet affected this asymmetry. It is uncertain whether postcopulatory advantages of large and protein-fed male medflies arise from female preferences or male dominance through coercion or force. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10458876 TI - Scanning and route selection in the jumping spider Portia labiata. AB - Jumping spiders Portia labiata were tested in the laboratory on three different kinds of detours. In one, both routes led to the lure. In the other variants, one of the routes had a gap, making that route impassable. When tested with only one complete route, Portia chose this route after visually inspecting both routes. An analysis of scanning showed that, at the beginning of the scanning routine, the spiders scanned both the complete and the incomplete route but that, by the end of the scanning routine, they predominantly scanned only the complete route. Two rules seemed to govern their scanning: (1) they would continue turning in one direction when scanning away from the lure along horizontal features of the detour route; and (2) when the end of the horizontal feature being scanned was reached, they would change direction and turn back towards the lure. These rules 'channelled' the spiders' scanning on to the complete route, and they then overwhelmingly chose to head towards the route they had fixated most while scanning. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10458877 TI - Possible causes of reproductive dominance during emergency queen rearing by honeybees. AB - When queenless honeybees, Apis mellifera, rear new queens, the relative frequencies of subfamilies found in the queen and worker brood are often very different, suggesting that certain subfamilies are reproductively dominant. At least two mechanisms could account for the observed differences in queen and worker broods. First, kin selection theory predicts that if honeybee workers are able to distinguish levels of relatedness, they should act nepotistically by favouring super-sisters over less-related half-sisters during emergency queen rearing. Alternatively, selection might result in royalty alleles that make their possessors more favoured for rearing as queens. Documented genetically based tendencies to rear queen or worker brood might interact with either of these mechanisms. To determine which of these effects might best explain reproductive dominance, we removed brood from the queenright section of one colony and offered it to the queenless section of the same colony and to three unrelated queenless colonies. We used two microsatellite loci to determine the paternity of queen and worker brood reared by these colonies. Variance in the proportions of subfamilies in queen and worker brood was greatest when the rearing bees were related to the brood. The results suggest that nepotistic interactions are more important than royalty alleles or other factors in causing reproductive dominance, but that there are complex interactions between the genotype of the nursing workers, and the genotypes of the larvae favoured for rearing as queens. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10458878 TI - Female mating strategy and male brood cannibalism in a sand-dwelling cardinalfish. AB - I investigated male and female sexual strategies of a cardinalfish, Apogon niger, which breeds in sandy areas at sporadic coral colonies. Males mouthbrood an egg mass received from one female at a time. Because of the lengthy mouthbrooding period, the interspawning interval was shorter for females than for males. Females moved between coral colonies to find mates more extensively than did males, and more frequently deserted mates after spawning. The females shortened their interspawning intervals by changing mates, especially in the late breeding season, when their mobility was highest. Their mobility was positively correlated with their disappearance rate, suggesting that mate search increases mortality. This may reduce competition between females for males, resulting in an unbiased rather than female-biased operational sex ratio. Males, on the other hand, practised filial cannibalism of entire broods, which might allow them partially to compensate for the lack of food during the mouthbrooding period. The reproductive loss entailed by filial cannibalism could be effectively offset if males remate soon afterwards. However, cannibal males took a long time to remate because few females were available. Filial cannibalism was less frequent than in a boulder-dwelling congener in which males have easy access to mates. Low mate availability may inhibit male A. niger from committing filial cannibalism. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10458879 TI - Coal tits, Parus ater, lose weight in response to chases by predators. AB - Theoretical models predict that birds should decrease their body mass in response to increased predation risk because lighter birds take off faster and are more manoeuvrable. We studied the effect of predation risk by chasing coal tits in large outdoor aviaries thus simulating an attempt to capture them. With this increase in predation risk, both perceived and actual, coal tits lost significantly more weight than in a control situation when they were not pursued. This pattern was attributable to a smaller gain in weight only during the day; nocturnal weight did not change in relation to diurnal predation risk. The lower daily weight gain was not consistent with predictions from models of interrupted foraging, but was consistent with predictions from risk adjustment models. Moreover, there was no difference in weight gain over 2-h periods that included a 1-h fast and those in which feeding was ad libitum, suggesting that coal tits could easily regain their body mass after a predator had interrupted their feeding. Our results therefore suggest that pursuit by predators leads to a decrease in the body mass of small birds. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10458880 TI - Operational sex ratio influences female preference and male-male competition in guppies. AB - Manipulation of the operational sex ratio (OSR) in guppies, Poecilia reticulata, causes changes in male-male competition and female mate choice. In this study OSR is defined as the number of sexually active males divided by the total number of sexually active adults of both sexes. The rate of male courtship displays decreased, and interference behaviours between males increased, at male-biased OSRs. The OSR influenced both copulatory tactics and postcopulatory guarding. All copulations followed sigmoid displays, except at an OSR of five males to one female where 60% of copulations occurred during sneak attempts. Compared with copulations that followed sigmoid displays, successful sneak copulations were followed by a shorter period of postcopulatory mate guarding and a shorter refractory period before males resumed courtship activities. Females preferred males with more orange colour whenever they had a choice, and the preference for orange colour was stronger with more male-biased OSRs. The OSR thus influences the presence, absence or relative importance of both female mate choice and male male competition which, in turn, should affect the evolution of secondary sexual traits. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10458881 TI - Stallion harassment and the mating system of horses. AB - Feral horse, Equus caballus, breeding groups, called bands, usually include one but sometimes up to five stallions. We found that mares were loyal to single stallion (SS) or multistallion (MS) bands or were social dispersers (maverick mares, Mv). The spacing and social behaviour of mares and stallions in single- and multistallion bands was measured. Indices of mare well-being were also measured including activity budgets (feeding: MS>SS=Mv; resting: MSSS), maternal effort in maintaining contact with foals (MS=Mv>SS), parasite levels in faeces (MS>Mv>SS), body condition (MS=Mv560 contiguous amino acid residues. Fgd2, like FGD1, contains adjacent RhoGEF and PH domains, a second carboxy-terminal PH domain, and a distinctive FYVE domain. Genomic PCR studies indicate some degree of conserved gene structure between Fgd2 and FGD1. Fgd2 transcripts are present in several diverse tissues and during mouse embryogenesis, suggesting a role in embryonic development. Genetic linkage and radiation hybrid mapping data show that Fgd2 and the human FGD2 ortholog map to syntenic regions of murine chromosome 17 and human chromosome 6p21.2, respectively. The observation that all FGD1 gene family members contain equivalent signaling domains and a conserved structural organization strongly suggests that these signaling domains form a canonical core structure for members of the FGD1 family of RhoGEF proteins. PMID- 10458913 TI - cDNA isolation, genomic structure, regulation, and chromosomal localization of human lung Kruppel-like factor. AB - Lung Kruppel-like factor (LKLF) is a zinc finger transcription factor critical for embryonic development. We have previously identified and isolated the mouse LKLF gene and examined its role using gene targeting. In this report, we describe the isolation and molecular characterization of the human homolog of murine LKLF. The human and mouse LKLF homologs exhibit an 85% nucleotide identity and share 90% amino acid similarity. Furthermore, the 5' sequence in the proximal promoter region and 3' untranslated region are also conserved between the two species. Of particular interest is the finding that while sequences in the proximal promoter have diverged between mouse and human, a region of 75 nucleotides is essentially identical. Site-directed mutagenesis in this region impairs the ability of the LKLF promoter to drive reporter gene expression, indicating that it represents a novel transcriptional element important in the regulation of LKLF gene expression. The activation domain is highly proline-rich and, similar to mouse LKLF, contains 22% proline residues. The human LKLF transcriptional unit is located in a genomic region of approximately 3 kb on chromosome 19p13.1. This region of chromosome 19 is known to contain genes involved in various human diseases. Like mouse LKLF, human LKLF consists of three exons that are interrupted by two small introns. The locations of intron/exon boundaries and splice sites are conserved between two homologs. Northern analysis shows that LKLF is expressed in lung in addition to heart, skeletal muscle, placenta, and pancreas. The isolation and chromosomal mapping of human LKLF will make it possible to initiate studies devoted to assess the involvement of this gene in human disease(s). PMID- 10458912 TI - A novel mitochondrial DNA-like sequence in the human nuclear genome. AB - We describe here a nuclear mitochondrial DNA-like sequence (numtDNA) that is nearly identical in sequence to a continuous 5842 bp segment of human mitochondrial DNA (mtDNA) that spans nucleotide positions 3914 to 9755. On the basis of evolutionary divergence among modern primates, this numtDNA molecule appears to represent mtDNA from a hominid ancestor that has been translocated to the nuclear genome during the recent evolution of humans. This numtDNA sequence harbors synonymous and nonsynonymous nucleotide substitutions relative to the authentic human mtDNA sequence, including an array of substitutions that was previously found in the cytochrome c oxidase subunit 1 and 2 genes. These substitutions were previously reported to occur in human mtDNA, but subsequently contended to be present in a nuclear pseudogene sequence. We now demonstrate their exclusive association with this 5842-bp numtDNA, which we have characterized in its entirety. This numtDNA does not appear to be expressed as a mtDNA-encoded mRNA. It is present in nuclear DNA from human blood donors, in human SH-SY5Y and A431 cell lines, and in rho(0) SH-SY5Y and rho(0) A431 cell lines that were depleted of mtDNA. The existence of human numtDNA sequences with great similarities to human mtDNA renders the amplification of pure mtDNA from cellular DNA very difficult, thereby creating the potential for confounding studies of mitochondrial diseases and population genetics. PMID- 10458914 TI - Cloning, expression profile, and genomic organization of the mouse STAP/A170 gene. AB - The preferential screening of cDNA libraries derived from the mouse osteoblastic cell line MC3T3-E1 has yielded a cDNA clone encoding a 442-amino-acid protein designated STAP (signal transduction and adaptor protein), which contains several motifs shared among transcription factors and adaptors such as a Zn-finger like motif, a proline-rich domain, and a PEST sequence. The amino acid sequence homology search also reveals that STAP is identical to a mouse oxidative stress protein, A170, and has 90% homology with a human p62 protein that binds to the tyrosine kinase p56(lck) SH2 domain. Northern blot analysis indicated a broad expression profile of STAP mRNA in various tissues and cell lines. In MC3T3-E1 cells, STAP mRNA was induced by treatment with TGF-beta, but not with BMP-2 or GDF-5. Analysis of the mouse STAP gene isolated from the genomic library revealed that the STAP gene spans a region of over 11 kb and comprises eight exons. The transcription start site was identified by primer extension analysis to be located 35 bp upstream from the translation initiation site. Sequencing analysis of the 5' flanking region of the STAP gene revealed multiple consensus motifs/sequences for several DNA binding transcription factors. The STAP gene had a TATA box, but no CCAAT box. Potential Sp1, AP-1, NF-E2, MyoD, and NF-kappaB binding sites were found in the 5' flanking region (1.4 kb) of the STAP gene. PMID- 10458915 TI - Identification, chromosomal assignment, and expression analysis of the human homeodomain-containing gene Orthopedia (OTP). AB - Homeodomain (HD) genes are helix-turn-helix transcription factors that play key roles in the specification of cell fates. In the central nervous system (CNS), HD genes not only position cells along an axis, but also specify cell migration patterns and may influence axonal connectivity. In an effort to identify novel HD genes involved in the development of the human CNS, we have cloned, characterized, and mapped the human homologue of the murine HD gene Orthopedia (Otp), whose product is found in multiple cell groups within the mouse hypothalamus, amygdala, and brain stem. Human cDNA and genomic libraries were screened with probes derived from mouse Otp sequences to find the human homologue, OTP. The deduced amino acid sequence of the open reading frame of the human cDNA is 99% homologous to mouse Otp and demonstrates a high degree of conservation when compared to sea urchin and Drosophila. OTP was mapped to human chromosome 5q13.3 using radiation hybrid panel mapping and fluorescence in situ hybridization. Flanking markers were identified from YAC clones containing OTP. A single putative OTP gene product was found in 17-week human fetal brain tissue by Western blot analysis using a novel polyclonal antibody raised against a conserved 13-amino-acid sequence at the C-terminus of the OTP protein. Expression in the developing human hypothalamus was confirmed by immunohistochemistry. PMID- 10458916 TI - Cloning and characterization of ZNF236, a glucose-regulated Kruppel-like zinc finger gene mapping to human chromosome 18q22-q23. AB - We report the cDNA cloning and characterization of ZNF236, a novel Kruppel-like zinc-finger gene initially identified by its glucose-regulated expression in human mesangial cells using mRNA differential display. Using the differential display fragment as a probe, we screened a human fetal kidney cDNA library and isolated several clones representing two differently spliced mRNA transcripts, designated ZNF236a and -b. Both transcripts were identical apart from the presence of an additional exon in ZNF236a that truncates the open reading frame. RT-PCR analysis confirmed the expression of both transcripts to be upregulated in human mesangial cells in response to elevated levels of d-glucose. ZNF236a and -b cDNAs encode polypeptides of 174 and 204 kDa, containing 25 and 30 C(2)H(2) zinc finger motifs, respectively. Northern blot analysis showed that ZNF236 is ubiquitously expressed in all human tissues tested. Expression levels were highest in skeletal muscle and brain, intermediate in heart, pancreas, and placenta, and lowest in kidney, liver, and lung. Southern zoo blot analysis indicated that ZNF236 is conserved in the genomes of all mammalian species tested, but not in yeast. The mapping of ZNF236 to human chromosome 18q22-q23, close to the IDDM6 locus, coupled with the glucose-regulated expression of the gene in human mesangial cells, suggests that ZNF236 may be a candidate gene for diabetic nephropathy. PMID- 10458917 TI - Co-production of staphylococcal enterotoxin A with toxic shock syndrome toxin-1 (TSST-1) enhances TSST-1 mediated mortality in a D-galactosamine sensitized mouse model of lethal shock. AB - It has previously been reported that staphylococcal enterotoxin A (SEA) is frequently co-expressed with toxic shock syndrome toxin-1 (TSST-1) in menstrual Toxic Shock Syndrome (MTSS)-associated Staphylococcus aureus. It was hypothesized that co-production of SEA and TSST-1 might yield a more virulent strain than one that produced TSST-1 but not SEA. To test this hypothesis, a TSST-1+/SEA- derivative of S. aureus RN3984 (TSST-1+/SEA+) was constructed by plasmid integration, and the isogenic pair were introduced into a D-galactosamine sensitized mouse model of lethal shock. At 72 h, 27 out of 30 (90%) mice inoculated with the parental strain died, as compared to 21 out of 30 (70%) mice inoculated with the isogenic derivative (P=0.05, Fisher's exact test; 1-tailed; 95% confidence limits, 0.80-20.80). Our results suggest that co-production of SEA with TSST-1 does enhance the ability of this strain of S. aureus to induce lethal shock in vivo. This enhanced virulence could be due to an additive or synergistic activity of the toxin combination on T cell proliferation and cytokine production in the animal model. PMID- 10458918 TI - Capsular hyaluronic acid of group A streptococci hampers their invasion into human pharyngeal epithelial cells. AB - Group A streptococci (GAS) cause various diseases, from uncomplicated noninvasive, to severe invasive infections. Capsular hyaluronic acid (HA) is known to resist phagocytosis, however, interaction between HA and epithelial cells have not been clearly understood. In this study, both HA-producing wild strains and HA-nonproducing mutants were employed to examine their invasiveness into confluent cultures of HEp-2, a nonphagocytic human epithelial cell line. Invasion of HEp-2 cells by GAS strains increased over time. The hasA gene encoding hyaluronate synthase of GAS strains was inactivated by allelic replacement. It was found that hasA-inactivated mutants were internalized into HEp-2 cells more efficiently than their parent strains under various conditions in terms of incubation time and inoculum size. Taken together, these findings indicate that GAS can be internalized into HEp-2 cells with considerably high frequencies and that the presence of HA of GAS decreased the invasion efficiency. PMID- 10458919 TI - Outer membrane vesicles of Porphyromonas gingivalis inhibit IFN-gamma-mediated MHC class II expression by human vascular endothelial cells. AB - Porphyromonas gingivalis is thought to be one of the major pathogenic organisms of adult periodontitis. Of the several virulence factors associated with the pathology it causes, evidence is now presented suggesting that outer membrane vesicles, which form from blebbing of the outer membrane, may also contribute to the pathogenesis of this bacterium. To evaluate this possibility, outer membrane vesicles were isolated from cultures of P. gingivalis and tested for their ability to promote inflammation and for their effects on the biosynthesis of E selectin and ICAM-1 adhesion molecules and MHC class II glycoproteins. The results indicate that these vesicles are capable of inducing acute inflammation characterized by the accumulation of a large number of neutrophils in the connective tissue. This cellular response corresponds to the vesicle-mediated biosynthesis and surface membrane expression of E-selectin and ICAM-1 by vascular endothelial cells. In contrast, IFN-gamma-dependent synthesis of MHC class II molecules was found to be inhibited by vesicles. Inhibition of HLA-DR expression occurred regardless of whether vesicles were added at the same time as, 24 h before, or 24 h after IFN-gamma stimulation of endothelial cells, suggesting that the inhibitory effects occur at both the membrane and intracellular level. These findings, taken together, indicate that P. gingivalis membrane vesicles are capable of inducing and regulating cellular responses involved in inflammation and initiation of acquired immunity. Membrane vesicles are composed of muramyl peptides, periplasmic proteins and outer membrane constituents. The combination of these components probably contribute to the immune regulatory functions reported herein. PMID- 10458920 TI - The Trichomonas vaginalis phenotypically varying P270 immunogen is highly conserved except for numbers of repeated elements. AB - The prominent and phenotypically variable immunogenic protein of Trichomonas vaginalis, termed P270, is present in all isolates. Most, if not all, patients make antibody to the DREGRD epitope contained in the 333 bp tandemly repeating element (TRE). The complete sequence of p270 of a fresh clinical isolate was recently derived (Musatovova and Alderete, Microb Pathogen 1998; 24: 223-39). We hypothesized that the size polymorphisms of P270 were due to the varied number of TREs that comprise a large, central portion of the gene. In this study, we analysed the p270 coding regions of ten representative isolates. It was determined also that the sequence of the TRE of different p270 genes shared > or =99% identity, and individual TREs of the same p270 gene showed them to have identical nucleotide sequences, affirming the highly-conserved nature of this element within each gene. The coding regions upstream and downstream of the central TREs were then generated by PCR amplification using specific primers. The PCR products corresponding to the 5' and 3'-end coding, non-repeat sequences were then subjected to restriction analyses, and the regions were highly conserved for all p270 genes. The complete sequence of two p270 genes showed > or = 99% identity of amino acids at the N- and C-terminal regions of p270, further reinforcing that the reported polymorphisms in Mr of P270 is due to the varying number of TREs and, therefore, the size of the TRE domain. In support of this hypothesis and during these analyses, one isolate, T. vaginalis T016, was discovered which possessed a p270 gene with only one partial repeat unit. Importantly, and as with all other p270 genes, transcription of this single repeat p270 gene in isolate T016 was confirmed. The start codon for the p270 T016 gene was preceded by the 12 nucleotide consensus Inr promoter-like sequence (TCATTTTTAATA) and possessed a putative transmembrane domain at the carboxy terminus. PMID- 10458921 TI - Genetic organization of the lipopolysaccharide O-antigen biosynthetic locus of Leptospira borgpetersenii serovar Hardjobovis. AB - Leptospiral LPS plays a critical role in immunity to leptospirosis and forms the basis for serological classification of Leptospira. However, neither the structure of leptospiral LPS nor the genetics of its biosynthesis have been elucidated. A probe derived from the rhamnose biosynthetic genes of L. interrogans serovar Copenhageni was used to identify the rfb locus of L. borgpetersenii serovar Hardjobovis. Chromosome walking and sequence analysis revealed an rfb locus spanning 36.7 kb, which consists of 31 ORFs transcribed in the same direction. Clusters of genes were identified which encode proteins related to enzymes involved in the biosynthesis of activated sugars including rhamnose. Additional ORFs in the locus encode glycosyltransferases for the assembly of the O-antigen subunit and integral membrane proteins for the transport of O-antigen subunits through the membrane and assembly into LPS. PMID- 10458923 TI - Red and green laser Doppler compared with capillary microscopy to assess skin microcirculation in the feet of healthy subjects. AB - Skin microvasculature consists of nutritive capillaries and subpapillary arteriolar and venular plexus connected by arteriolovenular anastomoses. Capillary perfusion is of paramount importance for skin viability. Recently a new combined laser Doppler instrument has become available, featuring a combination of near-infrared (RL; 780 nm) and green (GL; 543 nm) laser light sources. Theoretically, the red laser will penetrate deeper, whereas the green laser will read fairly superficially. This may enable differentiation between the more superficial, i.e., capillary, and the deeper skin layers. To test this hypothesis, the combined laser Doppler technique was compared with nail fold capillary microscopy in the feet of 10 healthy subjects. Seven males and 3 females with a median age of 26 (range 20-42) years and without arterial pathology were investigated. The laser Doppler (Periflux 4001, Perimed) was equipped with a special dual probe conducting both GL and RL. The probe was attached to the pulp of the big toe (with many AV-shunts) and to the nail fold, at the site where capillary microscopy was performed too. Laser Doppler and capillary perfusion was assessed at rest and during postocclusive reactive hyperemia. These measurements were performed both in the sitting and the supine positions to test the postural vasoconstriction response. Median resting and hyperemic skin perfusion with GL were lower (P < 0.01) than with the RL in both areas and positions, except for the resting value in the sitting position on the dorsum of the toe. Plantar perfusion was found significantly higher than dorsal perfusion only with the RL in the supine position (P < 0.01). GL and RL on the plantar, but not the dorsal, side showed a significantly decreased perfusion upon dependency (P < 0.05), both at rest and during hyperemia. In contrast, resting and peak capillary velocity did show a decrease on dependency (P < 0.05). Although the green laser measures a lower perfusion than does the red laser, which is likely to be derived from more superficial skin layers, it does not show a reactivity similar to that measured with capillary microscopy. Thus, it is questionable whether the green laser exclusively measures capillary perfusion. PMID- 10458922 TI - DNA amplification of nasopharyngeal aspirates in rats: a procedure to detect Pneumocystis carinii. AB - The diagnosis of Pneumocystis carinii pneumonia (PCP) requires invasive methods of bronchoalveolar lavage and lung biopsy. In this study, we examined efficacy of polymerase chain reaction (PCR) compared to Giemsa and silver ammoniacal staining to detect P. carinii in easily accessible extrapulmonary sites as well as lung. Samples were collected from lung, nasal and pharyngeal aspirates, gastric contents, urine and blood from dexamethasone treated or untreated virus-free Sprague-Dawley rats. All immunosuppressed lung samples were P. carinii positive by PCR analysis and both stains. Respectively DNA fragments of P. carinii were found in 93%, of nasal and 75% of pharyngeal aspirates, and 0% of sera, urine or gastric aspirates from immunosuppressed rats. However, no P. carinii cysts or trophozoites were found in nasal and pharyngeal aspirates (extrapulmonary sites) by silver ammoniacal or Giemsa staining. In comparison, none of the specimens from immunocompetent rats were PCR positive at any sites tested including the lungs. Therefore, PCR amplification products of nasal and pharyngeal aspirates showed that immunosuppressed rats with PCP can carry P. carinii DNA fragments in their upper respiratory tracts, but immunocompetent animals without PCP, are free of the organism and this suggests an approach to be investigated in humans with PCP. PMID- 10458925 TI - Elevated oxygen tension inhibits flow-induced dilation of skeletal muscle arterioles. AB - The purpose of this study was to determine if elevated oxygen tension affects flow-induced dilation of in situ skeletal muscle arterioles. Cremaster muscle preparations from Sprague-Dawley rats were superfused with physiological salt solution (PSS) and viewed via television microscopy. A video micrometer was used to measure changes in arteriolar diameter in response to the increase in flow produced by occlusion of a parallel branch from the parent arteriole. Erythrocyte velocity was measured with an optical Doppler velocimeter. The superfusate PO(2) was altered by changing the oxygen concentration of the equilibration gas between 0 and 21% O(2). Elevation of superfusate PO(2) to 10% O(2) and 21% O(2) significantly decreased arteriolar diameter compared to the control diameter during 0% O(2) superfusion. Increases in arteriolar diameter during parallel arteriolar occlusion were reduced as superfusate PO(2) was elevated. However, elevated PO(2) had no significant effect on erythrocyte velocity through the perfused daughter vessel, either prior to or during parallel occlusion. As a result, blood flow through the dilating vessel was reduced with elevated PO(2). As a result of the reduced arteriolar diameter, wall shear rate in the perfused daughter vessel was increased with elevated PO(2), both prior to and during parallel occlusion. These observations demonstrate that elevated PO(2) can override flow-induced vasodilation in the skeletal muscle microcirculation, leading to the persistence of an elevated wall shear rate. PMID- 10458924 TI - Water channel (aquaporin 1) expression and distribution in mammary carcinomas and glioblastomas. AB - The aquaporins represent a family of transmembrane water channel proteins that are widely distributed in various tissues throughout the body and play a major role in transcellular and transepithelial water movement. Most tumors have been shown to exhibit high vascular permeability and high interstitial fluid pressure, but the transport pathways for water within tumors remain unknown. In this study, we examined the distribution of the aquaporin 1 (AQP1) water channel protein in several types of transplanted tumor. Two mammary carcinomas, MCaIV and R3230AC, and three glioblastomas, HGL21, U87, and F98, were implanted in rats and mice. Two sites of implantation in rodents were chosen: a cranial window (CW) region and a subcutaneous (SC) region. Tissues were studied using immunoblot analysis and immunofluorescence staining. In the mammary carcinomas, AQP1 was localized in vascular structures; no differences between CW and SC regions were observed. Among the three glioblastomas, HGL21 and U87 exhibited similar AQP1 localization in vascular structures, whereas the center of F98 did not show vascular staining. Cell membranes of normal epithelial cells did not show AQP1 expression, while membranes of most tumor cells exhibited significant AQP1 expression. Interestingly, however, HGL21 and F98 in the CW locations showed no AQP1 expression on tumor cell membranes. These results show that the AQP1 water channel is heterogeneously expressed in tumor cells and their vasculature, and that the level of expression is determined not only by the specific cellular origin of the tumor, but also by the location of the tumor in the host animal. PMID- 10458926 TI - Effects of megakaryocyte growth and development factor (thrombopoietin) on liver endothelial cells in vitro. AB - Thrombopoietin (TPO) is the major regulator of growth and differentiation of megakaryocytes. Recent studies have shown that TPO also has activity on hematopoietic lineages other than megakaryocytes. However, little is known about the effects of TPO on nonhematopoietic cells expressing the TPO receptor, such as endothelial cells (EC). We have previously shown that specific murine liver EC (LEC-1) located in the hepatic sinusoids coexpress TPO and its receptor, c-mpl. Likewise, we showed that TPO has a proliferative effect on LEC-1. In this study, we have further examined the effects of TPO on other biological functions of LEC 1. Stimulation with TPO induced secretion of proinflammatory cytokines (i.e., IL 1beta, IL-6, TNF-alpha) from LEC-1. TPO-induced proliferation of LEC-1 was synergistically enhanced with the addition of TNF-alpha. TPO also induced the proliferation of LEC-1 in the presence of IFN-gamma, which alone inhibited the growth of these cells. TPO has no effect on other endothelial cell functions such as nitric oxide production and adhesion molecule expression. These observations establish a novel activity of TPO on murine liver endothelial cells in terms of inducing cytokine production by these cells. Our results suggest that this cytokine may act synergistically with other cytokines to induce LEC-1 proliferation. PMID- 10458927 TI - Hydraulic conductivity, albumin reflection and diffusion coefficients of pig mediastinal pleura. AB - Hydraulic conductivity (L), albumin reflection coefficient (sigma), and albumin diffusion coefficient (D) were measured across pig mediastinal pleura. The tissue (7 mm diameter) was bonded between two chambers. Flow (Q) of lactated Ringer solution between the chambers was measured in turn at driving pressures (DeltaP) of 2, 4, and 6 cm H(2)O. Value of L was proportional to the slope of the Q-DeltaP curve. Then Q was measured in turn at three albumin osmotic pressure differences (Deltapi equivalent to -1, -2, and -3 g/dl albumin concentration difference, DeltaC) with DeltaP constant at either 2, 3, 4, or 6 cm H(2)O. From Starling's equation, magnitude of sigma was the slope of the Q-Deltapi curve divided by the slope of the Q-DeltaP curve. We measured the diffusion of 0, 2, 5, and 10 g/dl albumin with tracer (125)I-albumin. Tracer mass (M) that diffused across the pleura was measured for 10 h using a well-type NaI(T1) detector. D was calculated from the slope of the M-time curve. Values of L averaged 2.0 x 10(-8) cm(3). s( 1). dyne(-1) (n = 23). Values of sigma were small (0.02-0.05) and sigma increased as flow increased 20-fold. D (n = 24) increased 3-fold from 2.7 x 10(-8) cm(2)/s as DeltaC increased from 0 to 10 g/dl. The small values of sigma indicated that mediastinal pleura provided little restriction to the passage of protein. PMID- 10458929 TI - Indomethacin-induced disturbances in villous microcirculation in the rat ileum. AB - Indomethacin is a widely used nonsteroidal antiphlogistic compound used-among others-for the treatment of rheumatoid arthritis in humans. Common side effects of indomethacin in the gastrointestinal tract include ulcerative lesions and petechial bleeding in the mucosa. In the rat, oral intake of indomethacin induces ulcerations in the mucosa of the stomach and, if administered systemically, edema, petechial bleeding, and ulcerations in the small intestine. In order to determine if systemic administration of indomethacin may induce changes in villous perfusion, we assessed the effect of indomethacin on erythrocyte velocity and the diameter of the main arteriole in the villi of the rat ileum. We found that indomethacin led to a significant decrease in mean arteriolar blood flow (6.3 +/- 0.8 vs 5.0 +/- 1.2 nl/min, means +/- SD) 7 days after administration. Mean diameter of the main arteriole remained unchanged (control, 7. 8 +/- 0.8 vs indomethacin, 7.0 +/- 0.9 microm). In conclusion, systemic application of indomethacin leads to a decrease in blood supply to the mucosa. We previously found an increase in villous perfusion when assessed 24 h after administration of indomethacin, accompanied by an increase in arteriolar diameter of the main arteriole. In summary, different regimens of application of indomethacin lead to varying observations in microcirculatory parameters in single villi. Further studies are required to identify the underlying mechanisms. PMID- 10458928 TI - Vascular endothelial growth factor-induced migration of vascular smooth muscle cells in vitro. AB - Angiogenesis is a complex process that includes recruitment and proliferation of mural cells-smooth muscle cells (SMC) and pericytes. Vascular endothelial growth factor (VEGF) has been shown to play an important role in angiogenesis and is an endothelial cell chemoattractant. In addition, certain VEGF isoforms have been implicated in the normal formation of smooth muscle cell-surrounded arteries. Because VEGF's role as a mural cell chemoattractant had not been explored, we examined the ability of VEGF to influence vascular SMC migration in vitro. A Boyden chamber migration assay demonstrated that VEGF (0-100 ng/ml) caused a dose dependent migration of SMC. VEGF did not cause proliferation of SMC. Reverse transcriptase-polymerase chain reaction analysis demonstrated the presence of both KDR and flt mRNA, two known VEGF receptors, in SMC cultures. Western blot analysis of SMC lysates confirmed these data, revealing bands migrating at approximately 200 kDa and slightly below 200 kDa consistent with KDR and flt. These observations demonstrate that VEGF receptors are present on SMC, and that VEGF can act as an SMC chemoattractant. PMID- 10458930 TI - A role of protein kinase C in the regulation of cytosolic phospholipase A(2) in bradykinin-induced PGI(2) synthesis by human vascular endothelial cells. AB - The purpose of this study was to elucidate the mechanism by which bradykinin (BK) enhances prostacyclin (PGI(2)) production in human umbilical vein endothelial cells (HUVEC). BK-induced enhancement of PGI(2) synthesis was observed in a dose- and time-dependent manner, and it also increased [Ca(2+)](i) followed by enhancement of cytosolic phospholipase A(2) (cPLA(2)) activity. The PKC inhibitors GF109203X and H7 attenuated the BK-induced increase in [Ca(2+)](i) and inhibited the BK-induced PGI(2) synthesis. Phorbol 12-myristate 13-acetate increased cPLA(2) activity and PGI(2) synthesis but failed to alter [Ca(2+)](i). BK increased cPLA(2) mRNA eightfold by 15 min, and this increase was inhibited by pretreatment with the PKC inhibitors. In response to cycloheximide pretreatment, cPLA(2) mRNA was superinduced. These results suggest that BK stimulates PGI(2) synthesis in HUVEC by activation of cPLA(2) by dual mechanisms: an elevation of [Ca(2+)](i) and a PKC-dependent pathway. Moreover, changes in calcium kinetics and expression of cPLA(2) mRNA may underlie the BK-induced PGI(2) enhancement in these cells. PMID- 10458931 TI - Thermodiffusion for continuous quantification of hepatic microcirculation- validation and potential in liver transplantation. AB - Hepatic microcirculation is a main determinant of reperfusion injury and graft quality in liver transplantation. Methods available for the quantification of hepatic microcirculation are indirect, are invasive, or preclude postoperative application. The aim of this study was the validation of thermodiffusion in a new modification allowing long-term use in the clinical setting. In six pigs Doppler flowmeters were positioned around the hepatic artery and portal vein for the measurement of total liver blood flow. Liver perfusion was quantified by thermodiffusion and compared to H(2) clearance as an established technique under baseline conditions, during different degrees of portal venous obstruction and during occlusion of the hepatic artery. Thermodiffusion measurements were recorded for five days postoperatively followed by histological evaluation of the hepatic puncture site. Perfusion data obtained by thermodiffusion were significantly correlated to H(2) clearance (r = 0.94, P < 0. 001) and to liver blood flow (r = 0.9, P < 0.05). The agreement between thermodiffusion and H(2) clearance was excellent (mean difference -2.1 ml/100 g/min; limits of agreement 12.5 and 8.3 ml/100 g/min). Occlusion of the portal vein or hepatic artery was immediately detected by thermodiffusion, indicating a decrease of perfusion by 64 +/- 7% or 27 +/- 5% of baseline, respectively. Perfusion values at baseline and during vascular occlusion were reproducible during the entire observation period. Histological changes of the liver tissue adjacent to the thermodiffusion probes were minute and did not influence long-term measurements. In vivo validation proved that enhanced thermodiffusion is a minimally invasive technique for the continuous, real-time quantification of hepatic microcirculation. Changes in liver perfusion can be safely detected over several days postoperatively. The implication for liver transplantation has led to the clinical application of thermodiffusion. PMID- 10458932 TI - Regular slow wave flowmotion in skeletal muscle is not determined by nitric oxide and endothelin. AB - In a previous study we showed that the generation of regular slow wave flowmotion (rSWFM, 1-3 cycles per minute) in skeletal muscle of anesthetized rats was related to local changes of arterial pressure and microcirculatory blood flow (MBF), which suggests an involvement of pressure- or flow-induced mechanisms. The present experiments were designed to test the role of flow-dependent endothelial autacoids, such as nitric oxide (NO) and endothelin, in the generation of SWFM. The effects of NO-donor sodium nitroprusside (SNP), the partly NO-dependent metabolite adenosine (ADO), the NO-synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME), and the mixed endothelin receptor blocker bosentan (BOS) were analyzed. MBF and rSWFM were assessed by laser Doppler flowmetry. rSWFM appeared in 7 out of 14 preparations after ADO (200 microg/kg/min), but not after SNP (100 microg/kg/min), L-NAME (30 mg/kg iv), and BOS (10 mg/kg iv). Its occurrence was associated with a significant decrease in arterial pressure to 50 +/- 3% (mean +/- SEM) of the baseline, provided that MBF was not enhanced. When given after induction of rSWFM by a 25% hemorrhage, SNP (50 microg/kg/min) totally abolished rSWFM and ADO (100 microg/kg/min) reduced rSWFM frequency from 2.17 +/- 0.08 to 1.72 +/- 0.08 cycles per minute (cpm) (P < 0.05), whereas the frequency was not affected by the other drugs. ADO, l-NAME (30 mg/kg iv), and BOS (10 mg/kg iv) lead to changes in rSWFM amplitude which showed a drug-independent negative correlation to changes in both MAP and MBF (R(2) = 0.61, multiple regression) in the ranges of 57-176% of MAP before drug application, and 72-120% of MBF, respectively. We conclude that NO and endothelin are not involved in the generation of rSWFM. Our findings strongly suggest that the activity of rSWFM depends on a reduction of vascular wall tension and is inhibited by SNP. PMID- 10458933 TI - Platelet-induced migration of smooth muscle cells under shear stress. PMID- 10458934 TI - Arteriolar growth in the postnatal rat heart. PMID- 10458935 TI - Red blood cell behavior at low flow rate in microvessels. PMID- 10458936 TI - Low-molecular-weight heparin inhibits neutrophil transmigration through endothelial cell monolayers. PMID- 10458937 TI - P-selectin-mediated rolling is a prerequisite for ICAM-1-independent firm adhesion in arterioles provoked by tumor necrosis factor-alpha in vivo. PMID- 10458938 TI - Nailfold capillary microscopy in human immunodeficiency virus-infected patients: a case-control study. PMID- 10458939 TI - The effect of intermittent normobaric hypoxia on vascularization of human myometrium. PMID- 10458940 TI - Three-dimensional MRI atlas of the human cerebellum in proportional stereotaxic space. AB - We have prepared an atlas of the human cerebellum using high-resolution magnetic resonance-derived images warped into the proportional stereotaxic space of Talairach and Tournoux. Software that permits simultaneous visualization of the three cardinal planes facilitated the identification of the cerebellar fissures and lobules. A revised version of the Larsell nomenclature facilitated a simple description of the cerebellum. This atlas derived from a single individual was instrumental in addressing longstanding debates about the gross morphologic organization of the cerebellum. It may serve as the template for more precise identification of cerebellar topography in functional imaging studies in normals, for investigating clinical-pathologic correlations in patients, and for the development of future probabilistic maps of the human cerebellum. PMID- 10458941 TI - Influence of motor activity on striatal dopamine release: A study using iodobenzamide and SPECT. AB - Pharmacologically induced dopamine release can influence the postsynaptic receptor binding of dopaminergic radioligands. This effect has recently been described using in vivo imaging methods and has been attributed to competition of radiotracers with the endogenous ligand. The present study examines the effect of a motor activation task on dopamine release and the consequences of this release on the binding of the selective D(2) receptor ligand (123)I-iodobenzamide (IBZM) to striatal dopamine D(2) receptors. Eight subjects were asked to write a text beginning immediately before IBZM injection and continuing for 30 min thereafter. Eighteen other subjects remained in a supine resting state during this period and served as a control group. All subjects were right handed. We hypothesized that the writing task would lead to an increase of dopamine release. The increased competition of the endogenous ligand with IBZM should lead to a decreased postsynaptic IBZM binding in the experimental group. Images were acquired and reconstructed identically and anatomically normalized to a computerized brain atlas. Regions of interest were drawn covering the striatum and three different reference regions. Ratios of striatal-to-reference-tissue radioactivity accumulation were calculated as semi-quantitative estimates of D(2) receptor binding potential. This decreased bilaterally, although right-sided significantly more than left, regardless of the choice of reference region. These data show that writing with the right hand compared to a supine resting state leads to a decrease of striatal IBZM accumulation. According to our primary hypothesis this reflects dopamine release. PMID- 10458942 TI - Cortical responses to sustained and divided attention in Alzheimer's disease. AB - Neuropsychological data suggests that divided attention is more impaired than sustained attention during the early phases of Alzheimer's disease. The purpose of the present study was to compare cerebral activation patterns during sustained and divided attention between Alzheimer patients and healthy elderly. The O-15 water PET activation method was used to map sustained and divided attention in 16 patients with Alzheimer's disease (mean age +/- SD: 68 +/- 5 years; MMSE: 11-25, mean +/- SD = 19.5 +/- 4.9) and in 16 healthy age-matched control subjects. After stereotactical normalization, voxel-by-voxel t statistics was used to assess the significance of activated brain areas and to compare activations between patients and control subjects. In the healthy elderly, sustained and divided attention both elicited activation of the right inferior parietal lobule, and the right middle frontal gyrus, whereas the anterior cingulate gyrus was activated during sustained attention only. Only medial frontal structures (Brodmann Area (BA) 32/34) were activated in Alzheimer patients, and both frontal (BA-10), posterior cingulate (BA-23/31), and subcortical sites were deactivated. Compared to the healthy elderly, the activations in the patients of the right medial (BA-11) superior (BA-10) and inferior (BA-47) frontal gyri, the right middle temporal (BA 20), and the left lingual (BA-17) gyri were significantly reduced. More cortical sites differed statistically between Alzheimer patients and control subjects during divided than during sustained attention. The activation pattern elicited by attention supports the neuropsychological data that divided attention is more impaired than sustained attention in early Alzheimer's disease. PMID- 10458943 TI - Plurality and resemblance in fMRI data analysis. AB - We apply nine analytic methods employed currently in imaging neuroscience to simulated and actual BOLD fMRI signals and compare their performances under each signal type. Starting with baseline time series generated by a resting subject during a null hypothesis study, we compare method performance with embedded focal activity in these series of three different types whose magnitudes and time courses are simple, convolved with spatially varying hemodynamic responses, and highly spatially interactive. We then apply these same nine methods to BOLD fMRI time series from contralateral primary motor cortex and ipsilateral cerebellum collected during a sequential finger opposition study. Paired comparisons of results across methods include a voxel-specific concordance correlation coefficient for reproducibility and a resemblance measure that accommodates spatial autocorrelation of differences in activity surfaces. Receiver-operating characteristic curves show considerable model differences in ranges less than 10% significance level (false positives) and greater than 80% power (true positives). Concordance and resemblance measures reveal significant differences between activity surfaces in both data sets. These measures can assist researchers by identifying groups of models producing similar and dissimilar results, and thereby help to validate, consolidate, and simplify reports of statistical findings. A pluralistic strategy for fMRI data analysis can uncover invariant and highly interactive relationships between local activity foci and serve as a basis for further discovery of organizational principles of the brain. Results also suggest that a pluralistic empirical strategy coupled formally with substantive prior knowledge can help to uncover new brain-behavior relationships that may remain hidden if only a single method is employed. PMID- 10458944 TI - Spectroscopic analysis of changes in remitted illumination: the response to increased neural activity in brain. AB - Imaging of neural activation has been used to produce maps of functional architecture and metabolic activity. There is some uncertainty associated with the sources underlying the intrinsic signals. It has been reported that following increased neural activity there was little increased oxygen consumption ( approximately 5%), although glucose consumption increased by approximately 50%. The research we describe uses a modification of the Beer-Lambert Law called path length scaling analysis (PLSA) to analyze the spectra of the hemodynamic and metabolic responses to vibrissal stimulation in rat somatosensory cortex. The results of the PLSA algorithm were compared with those obtained using a linear spectrographic analysis method (we refer to this as LMCA). There are differences in the results of the analysis depending on which of the two algorithms (PLSA or LMCA) is used. Using the LMCA algorithm, we obtain results showing an increase in the volume of Hbr at approximately 2 s, following onset of stimulation but no complementary decrease in oxygenated haemoglobin (HbO(2)). These results are similar to a previous report. In contrast, after using the PLSA algorithm, the time series of the chromophore changes shows no evidence for an increase in the volume of deoxygenated haemoglobin (Hbr). However, after further analysis of the time series from the PLSA using general linear models (GLM) to remove contributions from low frequency baseline oscillations, both the HbO(2) and Hbr times series of the response to stimulation were found to be biphasic with an early decrease in saturation peaking approximately 1 s after onset of stimulation followed by a larger increase in saturation peaking at approximately 3 s. Finally, following the PLSA-then-GLM analysis procedure, we do not find convincing evidence for an increase in cytochrome oxidation following stimulation, though we demonstrate the PLSA algorithm to be capable of disassociating changes in cytochrome oxidation state from changes in hemoglobin oxygenation. PMID- 10458945 TI - A developmental functional MRI study of spatial working memory. AB - Functional magnetic resonance imaging (fMRI) was used to examine patterns of cortical activity in children during performance of a spatial working memory task. Six children (8-10 years) and six adults (19-26 years) searched a linear array of four boxes for the appearance of a dot. In the visual blocks, participants made no response. In the motor blocks, participants were instructed to indicate the location of the dot on each trial using a button-press response. In the working memory blocks, participants were instructed to indicate at which location the dot had appeared 1 or 2 trials previously. Both children and adults showed activity in the left precentral and postcentral gyri, as well as the right cerebellum for the motor condition as compared to the visual condition. Comparison of the memory and motor conditions revealed reliable activity in the right superior frontal gyrus (BA 8), right dorsolateral prefrontal cortex (BA 10/46), right superior parietal cortex, and bilateral inferior parietal cortex for both adults and children. These results suggest that spatial working memory tasks activate very similar cortical regions for school-age children and adults. The findings differ from previous imaging studies of nonspatial working memory tasks in that the prefrontal activations observed in the current work tend to be more dorsal. Results are discussed in light of the significant behavioral performance differences observed between child and adult participants. PMID- 10458946 TI - Activation of the remaining hemisphere following stimulation of the blind hemifield in hemispherectomized subjects. AB - We used functional magnetic resonance imaging (fMRI) to investigate the neural substrates mediating residual vision in the "blind" hemifield of hemispherectomized patients. The visual stimuli were semicircular gratings moving in opposite directions on a dynamic random-dot background. They were specifically constructed to eliminate intra- and extraocular light scatter and optimize the activation of extrastriate cortical areas and their subcortical relays. Multislice T2*-weighted gradient echo (GE) echoplanar imaging (EPI) images (TR/TE = 4 s/45 ms, flip angle 90 degrees ) were acquired during activation and baseline visual stimulation. An activation minus baseline subtraction was performed, and the acquired t statistic map transformed into the stereotaxic coordinate space of Talairach and Tournoux. In seven normal control subjects, right hemifield stimulation produced significant activation foci in contralateral V1/V2, V3/V3A, VP, and V5 (MT). Significant activation was also produced in homologous regions of the right occipital lobe with left hemifield stimulation. Stimulation of the intact hemifield in hemispherectomized patients resulted in activation of similar areas exclusively within the contralateral hemisphere. Stimulation of the anopic hemifield produced statistically significant activation in the ipsilateral occipital lobe (putative area V5 or MT) and areas V3/V3A in the only subject with blindsight. We conclude that the remaining hemisphere may contribute to residual visual functions in the blind hemifield of hemispherectomized patients, possibly through the collicular-pulvinar route since the activated areas are known to receive their afferents from these subcortical nuclei. PMID- 10458947 TI - Proceedings of the National Symposium on Medical and Public Health Response to Bioterrorism. Arlington, Virginia, USA. February 16-17, 1999. PMID- 10458949 TI - The emerging threat of bioterrorism. PMID- 10458948 TI - Bioterrorism: how prepared are we? PMID- 10458950 TI - View from the Hill: Congressional efforts to address bioterrorism. PMID- 10458951 TI - Finding the right balance against bioterrorism. PMID- 10458952 TI - Historical trends related to bioterrorism: An empirical analysis. PMID- 10458953 TI - The threat of biological attack: why concern now? PMID- 10458954 TI - Nuclear blindness: An overview of the biological weapons programs of the former Soviet Union and Iraq. PMID- 10458955 TI - Aum Shinrikyo: once and future threat? PMID- 10458956 TI - The prospect of domestic bioterrorism. PMID- 10458957 TI - Potential biological weapons threats. PMID- 10458958 TI - Epidemiology of bioterrorism. PMID- 10458959 TI - Vaccines in civilian defense against bioterrorism. PMID- 10458960 TI - Vaccines, pharmaceutical products, and bioterrorism: challenges for the U.S. Food and Drug Administration. PMID- 10458961 TI - Smallpox: clinical and epidemiologic features. PMID- 10458962 TI - Smallpox: An attack scenario. PMID- 10458963 TI - Aftermath of a hypothetical smallpox disaster. PMID- 10458964 TI - Clinical and epidemiologic principles of anthrax. PMID- 10458965 TI - Anthrax: A possible case history. PMID- 10458966 TI - Applying lessons learned from anthrax case history to other scenarios. PMID- 10458967 TI - Addressing bioterrorist threats: where do we go from here? PMID- 10458968 TI - The surveillance of vero cytotoxin-producing Escherichia coli O157 in Wales, 1990 to 1998. AB - Population-based surveillance for Vero cytotoxin-producing Escherichia coli (VTEC) O157 has been carried out in Wales since 1990. The annual incidence has remained stable during the 9-year period (mean: 1.6 cases per 100,000 population); the rate is highest in children younger than 5 years of age. Blood in the stool is reported in fewer than half the cases, indicating the importance of screening all fecal specimens for VTEC O157. PMID- 10458969 TI - Chlorine disinfection of recreational water for Cryptosporidium parvum. AB - We examined the effects of chlorine on oocyst viability, under the conditions of controlled pH and elevated calcium concentrations required for most community swimming pools. We found that fecal material may alter the Ct values (chlorine concentration in mg/L, multiplied by time in minutes) needed to disinfect swimming pools or other recreational water for Cryptosporidium parvum. PMID- 10458970 TI - Cyclospora cayetanensis among expatriate and indigenous populations of West Java, Indonesia. AB - From January 1995 through July 1998, we investigated the occurrence of Cyclospora cayetanensis infection associated with gastrointestinal illness or diarrhea in foreign residents and natives of West Java, Indonesia. We found that C. cayetanensis was the main protozoal cause of gastrointestinal illness and diarrhea in adult foreign residents during the wet season. The parasite rarely caused illness in the indigenous population or in children. PMID- 10458971 TI - The first major outbreak of dengue hemorrhagic fever in Delhi, India. AB - India An outbreak of dengue hemorrhagic fever/dengue shock syndrome (DHS/DSS) occurred in 1996 in India in and near Delhi. The cause was confirmed as dengue virus type 2, by virus cultivation and indirect immunofluorescence with type specific monoclonal antibodies. This is the largest such outbreak reported from India, indicating a serious resurgence of dengue virus infection. PMID- 10458972 TI - Addressing the potential threat of bioterrorism--value added to an improved public health infrastructure. PMID- 10458973 TI - Current status of smallpox vaccine. PMID- 10458974 TI - West Nile fever in Czechland. PMID- 10458975 TI - Ofloxacin-resistant Vibrio cholerae O139 in Hong Kong. PMID- 10458976 TI - Plant pathology and public health. PMID- 10458977 TI - Pet-associated zoonoses. PMID- 10458978 TI - Avirulence and resistance genes in the Cladosporium fulvum-tomato interaction. AB - The fungus Cladosporium fulvum infects tomato and secretes various proteins that are recognized by resistant plants that respond with a hypersensitive response. Strains of the fungus that escape recognition by tomato are virulent. Resistance genes in tomato, either directly or indirectly involved in recognition of the fungal proteins, encode extracellular membrane-anchored, leucine-rich repeat proteins, which occur in gene clusters. Much progress has been made in our understanding of the evolution of recognitional specificities in the host plant. PMID- 10458979 TI - Immunity to Candida albicans: Th1, Th2 cells and beyond. AB - Resistance to Candida albicans infection in mice results from the development of T helper (Th) type 1 cell responses. Cytokines produced by Th1 cells activate macrophages and neutrophils to a candidacidal state. The development of Th2 responses underlines susceptibility to infection, because cytokines produced by Th2 cells inhibit Th1 development and deactivate phagocytic effector cells. With the recognition of the reciprocal influences between innate and adaptive Th immunity, it appears that the coordinated action of these two lines of immune defense is required to efficiently oppose the infectivity of the fungus and to determine its lifelong commensalism at the mucosal level. PMID- 10458980 TI - Immune deficiency, immune silencing, and clonal exhaustion of T cell responses during viral infections. AB - Analyses of the complex regulatory networks leading to T cell survival, death, and immune deficiency have been aided in the past year by the dramatic development of new technologies to identify T cells and assess T cell function. These new techniques have shown that functional inactivation and apoptotic elimination of both virus-specific and non-virus-specific T cell populations mold T cell responses to viral infections. PMID- 10458981 TI - Cell wall dynamics in yeast. AB - The yeast Saccharomyces cerevisiae is the first fungus for which the structure of the cell wall is known at the molecular level. It is a dynamic and highly regulated structure. This is vividly illustrated when the cell wall is damaged and a salvage pathway becomes active, resulting in compensatory changes in the wall. PMID- 10458982 TI - Virulence factors of Entamoeba histolytica. AB - Recent studies have increased our knowledge of Entamoeba histolytica cell biology and gene regulation. In the ameba, dominant-negative mutations in the Gal/GalNAc lectin affect adhesion and cytolysis, whereas mutations in meromyosin affect cytoskeletal function. Studying these mutant proteins has improved our understanding of the role of these proteins in E. histolytica virulence. The characterization of the CP5 cysteine protease and the induction of apoptosis in host target cells has led to a better comprehension of the mechanisms by which trophozoites can lyse target cells. PMID- 10458983 TI - Genome projects, genetic analysis, and the changing landscape of malaria research. AB - Genome analysis of the Plasmodium falciparum malaria parasite already is identifying genes relevant to therapeutic- and vaccine-related research. The genetic blueprint of P. falciparum will ultimately need to be understood at multiple levels of an integrated system and will provide a detailed account of the life processes of the parasite and of the devastating disease it causes. PMID- 10458984 TI - Recent developments in the biology of respiratory syncytial virus: are vaccines and new treatments just round the corner? AB - Respiratory syncytial virus (RSV) is the major cause of infantile bronchiolitis, and is an important pathogen in the elderly and in the developing world. The production of full length cDNA clones now allows precise genetic engineering of RSV, while knowledge of the immunopathogenesis of augmented disease gives hope that effective vaccines will soon be developed. PMID- 10458986 TI - Host and viral genetics of chronic infection: a mouse model of gamma-herpesvirus pathogenesis. AB - A general association of human and primate lymphotropic herpesviruses (gamma herpesviruses) with the development of lymphomas, as well as other tumors, especially in immunocompromised hosts, has been well documented. The lack of relevant small animal models for human gamma-herpesviruses has impeded progress in understanding the role of these viruses in the development of chronic disease. Recent research characterizing infection of inbred strains of mice with a murine gamma-herpesvirus, gamma-herpesvirus 68 (gammaHV68), is providing insights into viral and host factors involved in the establishment and control of chronic gamma herpesvirus infection. PMID- 10458985 TI - Signal transduction cascades regulating mating, filamentation, and virulence in Cryptococcus neoformans. AB - Cryptococcus neoformans is a basidiomycetous fungal pathogen that infects the central nervous system. The organism has a defined sexual cycle involving mating between haploid MATalpha and MATa cells. Recent studies have revealed signaling cascades that coordinately regulate differentiation and virulence of C. neoformans. One signaling cascade involves a conserved G-protein alpha subunit and cAMP, and senses nutrients during mating and virulence. The second is a conserved mitogen activated protein (MAP) kinase cascade that senses pheromone during mating, and also regulates haploid fruiting and virulence. Interestingly, some of the MAP kinase components are encoded by the MAT locus itself, which may explain the unique association of the MATalpha locus with physiology and virulence. PMID- 10458987 TI - Viruses as triggers of autoimmunity: facts and fantasies. AB - Autoimmunity has been proposed as the cause of several human chronic inflammatory diseases, and recent animal studies show that viruses can induce autoimmune disease. These studies demonstrate how viruses might misdirect the immune system, and here we discuss critically the evidence that similar phenomena may lead to human disease. PMID- 10458989 TI - Adhesins in Candida albicans. AB - The adherent properties of Candida albicans blastoconidia and germ tubes have long been appreciated, but little is known about the mechanisms responsible for adherence. Recently, three genes, ALA1, ALS1 and HWP1, encoding proteins with adherent properties and motifs consistent with linkage to the beta-1, 6-glucan of C. albicans cell walls have provided insight into the topology of protein adhesins. Hwp1, a developmentally regulated adhesin of germ tubes and hyphae, attaches to buccal epithelial cells by an unconventional, transglutaminase mediated mechanism of adhesion. PMID- 10458988 TI - Antigenic variation in Plasmodium falciparum: mechanisms and consequences. AB - In the past year, the major advances in malaria antigenic variation have been concerned with the transcription and switching of variant antigen genes, and the functional expression of regions of the major variant antigen. Also, new variant gene families have been discovered as a result of the Malaria Genome Project. PMID- 10458990 TI - Inhibition of host cell signal transduction by Leishmania: observations relevant to the selective impairment of IL-12 responses. AB - Leishmania parasites are able to delay the onset of cell-mediated immunity by selectively impairing the ability of infected macrophages to produce interleukin (IL)-12. Leishmania infection arrests the JAK/STAT-mediated signal transduction involved in activation of the IL-12 p40 promoter; the phosphorylation defects may be initiated by ligation of the phagocyte receptors used by these organisms to gain entry into the host cell. PMID- 10458991 TI - Initial and innate responses to viral infections--pattern setting in immunity or disease. AB - Host responses to infectious challenges include initial events elicited directly by agent structures distinct from host determinants, activation of innate immune system components by the products of initial events, and the shaping of downstream adaptive immunity by these initial/innate responses. The picture emerging from viral infections is that viral structures interact with intracellular signaling pathways to induce expression of the type 1 interferons, IFN-alpha/beta. In addition to mediating direct antiviral effects, these cytokines play dominant roles in regulating innate and adaptive immune responses to infection. In particular, IFN-alpha/beta acts to inhibit interleukin-12 (IL 12) expression and IL-12 activation of innate natural killer (NK) cell IFN-alpha production, while inducing NK cell cytotoxicity and proliferation, and promoting adaptive T cell IFN-alpha responses. Although certain viral infections do elicit initial/innate IL-12 and NK-cell-produced IFN-alpha, endogenous IFN-alpha/beta also controls the magnitudes of these responses. Thus, the pathways activated, to dominantly regulate innate and adaptive immune responses during viral infections, are being defined. PMID- 10458992 TI - Viral-induced neurodegenerative disease. AB - Viral etiology has been postulated in a variety of neurological diseases in humans, including multiple sclerosis. Several experimental animal models of viral induced neurodegenerative disease provide insight into potential host- and pathogen-dependent mechanisms involved in the disease process. Two such mouse models are the Theiler's murine encephalomyelitis virus (TMEV) infection and mouse hepatitis virus (MHV) infection. PMID- 10458993 TI - Origin, targeting, and function of the apicomplexan plastid. AB - The discovery of a plastid in Plasmodium, Toxoplasma and related protozoan parasites provides a satisfying resolution to several long-standing mysteries: the mechanism of action for various surprisingly effective antibiotics; the subcellular location of an enigmatic 35 kb episomal DNA; and the nature of an unusual intracellular structure containing multiple membranes. The apicomplexan plastid highlights the importance of lateral genetic transfer in evolution and provides an accessible system for the investigation of protein targeting to secondary endosymbiotic organelles. Combining molecular genetic identification of targeting signals with whole genome analysis promises to yield a complete picture of organellar metabolic pathways and new targets for drug design. PMID- 10458994 TI - Role of lectins (and rhizobial exopolysaccharides) in legume nodulation. AB - The lectin recognition hypothesis proposes that plant lectins mediate specificity in the Rhizobium-legume symbiosis. Although the hypothesis was developed eight years before nod genes were identified in rhizobia and sixteen years before Nod factor was shown to be a major determinant of host specificity, experiments performed recently using transgenic lectin plants support its main tenets. PMID- 10458995 TI - Genealogy of legume-Rhizobium symbioses. AB - Accumulating evidence suggests that lateral transfer of nodulation capacity is an important driving force in symbiotic evolution. As a consequence, many distantly related soil bacteria have acquired the capacity to invade plants and fix nitrogen within them. In addition to these proteins required for bacteroid development and nitrogen fixation, core symbiotic competence seems to require flavonoids, NodD proteins, lipochitooligosaccharidic Nod-factors, extra-cellular polysaccharides, as well as various exported proteins. Plants respond to different levels and combinations of these substances in species specific ways. After contact has been initiated by flavonoids and NodD proteins, constant signal exchange fine-tunes these symbiotic demands, especially to overcome defence reactions. PMID- 10458996 TI - Genes controlling expression of defense responses in Arabidopsis. AB - In the past year, two regulatory defense-related genes, EDS1l and COl1, have been cloned. Several other genes with regulatory functions have been identified by mutation, including DND1, PAD4, CPR6, and SSl1. It has become clear that jasmonate signaling plays an important role in defense response signaling, and that the jasmonate and salicylic acid signaling pathways are interconnected. PMID- 10458997 TI - Plant-insect interactions. AB - Recent research shows partially overlapping signal transduction pathways controlling responses to wounding, insects, and pathogens. Chemical and behavioral assays show that plants release herbivore-specific volatiles, and that parasitic wasps can distinguish between these emission patterns. QTL mapping and candidate gene studies are beginning to identify polymorphic resistance genes, and ecological analyses provide information on the physiological and fitness costs of resistance. Such multidisciplinary approaches can elucidate the physiological causes and ecological consequences of plant-herbivore interactions. PMID- 10458998 TI - Early events in host-pathogen interactions. AB - Research focused on early events in host-pathogen interactions has provided new insights into fundamental aspects of microbial pathogenicity and plant responses. Considerable progress has been made in understanding regulation of the delivery of pathogenicity determinants from bacteria into plant cells, signal cascades involved in fungal pathogenicity, the co-ordinating role of the plant cytoskeleton in plant defence and calcium flux as a primary signalling function during the hypersensitive reaction. PMID- 10458999 TI - Functional analysis of plant disease resistance genes and their downstream effectors. AB - Plant disease resistance (R) genes encode proteins that both determine recognition of specific pathogen-derived avirulence (Avr) proteins and initiate signal transduction pathways leading to complex defense responses. Recent developments suggest that recognition specificity of R proteins is determined by either a protein kinase domain or by a region consisting of leucine-rich repeats. R genes conferring resistance to bacterial, viral, and fungal pathogens appear to use multiple signaling pathways, some of which involve distinct proteins and others which converge upon common downstream effectors. Manipulation of R genes and their signaling pathways by transgenic expression is a promising strategy to improve disease resistance in plants. PMID- 10459000 TI - Defence signalling pathways in cereals. AB - The combination of mutational and molecular studies has shed light on the role of reactive oxygen intermediates and programmed cell death in cereal disease resistance mechanisms. Rice Rac1 and barley Rar1 represent conserved disease resistance signalling genes, which may have related functions in animals. The analysis of non-pathogenic Magnaporthe grisea mutants may provide novel tools to study host defence pathways. PMID- 10459001 TI - Role of active oxygen species and NO in plant defence responses. AB - Research in the area of active oxygen species is going through a reflective stage. There is controversy whether multiple mechanisms for active oxygen species generation exist and some data may need reassessing since the discovery of a role for NO in defence responses. Important work concerning upstream and downsteam signalling in this area is emerging, and the stage is set for approaches utilising transgenic knockouts and mutants to resolve many questions. PMID- 10459002 TI - Plant nematode resistance genes. AB - Root-knot and cyst nematodes cause severe damage to crops throughout the world. Genes conferring resistance against nematodes have been identified in many plant species and several of these have been, or soon will be, cloned. Nematode biotypes that can infect resistant plants have been identified. Investigation of cloned resistance genes and of virulent nematodes is likely to lead to improved host resistance. PMID- 10459003 TI - Probing plant cell structure and function with viral movement proteins. AB - Virus-encoded movement proteins are the principal strategy by which all plant viruses counter the primary physical defense of the plant to infection - the cell wall - to produce systemic infection and disease. Our understanding of how these proteins act at the molecular and cellular level has increased enormously in the past decade and ushered in an exciting new era of plant virology as an approach to investigating plant cell structure and function. The earliest studies focused on how movement proteins interacted with plasmodesmata, and were an important element in demonstrating the dynamic nature of these intercellular channels. Current efforts are focused on the role of movement proteins in coordinating the replication of viral genomes and the vectorial movement of the progeny genomes through the infected cell, as well as into adjacent cells. Movement proteins are thus providing unique approaches to unravel the fundamental mechanisms by which macromolecular transport is directed and integrated within and between plant cells. PMID- 10459004 TI - Medicago truncatula--a model in the making! PMID- 10459006 TI - Extranuclear apoptosis. The role of the cytoplasm in the execution phase. PMID- 10459005 TI - Employment of the epidermal growth factor receptor in growth factor-independent signaling pathways. PMID- 10459007 TI - Mcm2, but not RPA, is a component of the mammalian early G1-phase prereplication complex. AB - Previous experiments in Xenopus egg extracts identified what appeared to be two independently assembled prereplication complexes (pre-RCs) for DNA replication: the stepwise assembly of ORC, Cdc6, and Mcm onto chromatin, and the FFA-1 mediated recruitment of RPA into foci on chromatin. We have investigated whether both of these pre-RCs can be detected in Chinese hamster ovary (CHO) cells. Early and late-replicating chromosomal domains were pulse-labeled with halogenated nucleotides and prelabeled cells were synchronized at various times during the following G1-phase. The recruitment of Mcm2 and RPA to these domains was examined in relation to the formation of a nuclear envelope, specification of the dihydrofolate reductase (DHFR) replication origin and entry into S-phase. Mcm2 was loaded gradually and cumulatively onto both early- and late-replicating chromatin from late telophase throughout G1-phase. During S-phase, detectable Mcm2 was rapidly excluded from PCNA-containing active replication forks. By contrast, detergent-resistant RPA foci were undetectable until the onset of S phase, when RPA joined only the earliest-firing replicons. During S-phase, RPA was present with PCNA specifically at active replication forks. Together, our data are consistent with a role for Mcm proteins, but not RPA, in the formation of mammalian pre-RCs during early G1-phase. PMID- 10459008 TI - The ribosome regulates the GTPase of the beta-subunit of the signal recognition particle receptor. AB - Protein targeting to the membrane of the ER is regulated by three GTPases, the 54 kD subunit of the signal recognition particle (SRP) and the alpha- and beta subunit of the SRP receptor (SR). Here, we report on the GTPase cycle of the beta subunits of the SR (SRbeta). We found that SRbeta binds GTP with high affinity and interacts with ribosomes in the GTP-bound state. Subsequently, the ribosome increases the GTPase activity of SRbeta and thus functions as a GTPase activating protein for SRbeta. Furthermore, the interaction between SRbeta and the ribosome leads to a reduction in the affinity of SRbeta for guanine nucleotides. We propose that SRbeta regulates the interaction of SR with the ribosome and thereby allows SRalpha to scan membrane-bound ribosomes for the presence of SRP. Interaction between SRP and SRalpha then leads to release of the signal sequence from SRP and insertion into the translocon. GTP hydrolysis then results in dissociation of SR from the ribosome, and SRP from the SR. PMID- 10459009 TI - The developmental role of warthog, the notch modifier encoding Drab6. AB - The warthog (wrt) gene, recovered as a modifier for Notch signaling, was found to encode the Drosophila homologue of rab6, Drab6. Vertebrate and yeast homologues of this protein have been shown to regulate Golgi network to TGN trafficking. To study the function of this protein in the development of a multicellular organism, we analyzed three different warthog mutants. The first was an R62C point mutation, the second a genomic null, and the third was an engineered GTP bound form. Our studies show, contrary to yeast, that the Drosophila homologue of rab6 is an essential gene. However, it has limited effects on development beyond the larval stage. Only the mechanosensory bristles on the head, notum, and scutellum are affected by warthog mutations. We present models for the modifying effect of Drab6 on Notch signaling. PMID- 10459010 TI - Electrospray ionization tandem mass spectrometry (ESI-MS/MS) analysis of the lipid molecular species composition of yeast subcellular membranes reveals acyl chain-based sorting/remodeling of distinct molecular species en route to the plasma membrane. AB - Nano-electrospray ionization tandem mass spectrometry (nano-ESI-MS/MS) was employed to determine qualitative differences in the lipid molecular species composition of a comprehensive set of organellar membranes, isolated from a single culture of Saccharomyces cerevisiae cells. Remarkable differences in the acyl chain composition of biosynthetically related phospholipid classes were observed. Acyl chain saturation was lowest in phosphatidylcholine (15.4%) and phosphatidylethanolamine (PE; 16.2%), followed by phosphatidylserine (PS; 29.4%), and highest in phosphatidylinositol (53.1%). The lipid molecular species profiles of the various membranes were generally similar, with a deviation from a calculated average profile of approximately +/- 20%. Nevertheless, clear distinctions between the molecular species profiles of different membranes were observed, suggesting that lipid sorting mechanisms are operating at the level of individual molecular species to maintain the specific lipid composition of a given membrane. Most notably, the plasma membrane is enriched in saturated species of PS and PE. The nature of the sorting mechanism that determines the lipid composition of the plasma membrane was investigated further. The accumulation of monounsaturated species of PS at the expense of diunsaturated species in the plasma membrane of wild-type cells was reversed in elo3Delta mutant cells, which synthesize C24 fatty acid-substituted sphingolipids instead of the normal C26 fatty acid-substituted species. This observation suggests that acyl chain-based sorting and/or remodeling mechanisms are operating to maintain the specific lipid molecular species composition of the yeast plasma membrane. PMID- 10459011 TI - Phosphoinositide-AP-2 interactions required for targeting to plasma membrane clathrin-coated pits. AB - The clathrin-associated AP-2 adaptor protein is a major polyphosphoinositide binding protein in mammalian cells. A high affinity binding site has previously been localized to the NH(2)-terminal region of the AP-2 alpha subunit (Gaidarov et al. 1996. J. Biol. Chem. 271:20922-20929). Here we used deletion and site- directed mutagenesis to determine that alpha residues 21-80 comprise a discrete folding and inositide-binding domain. Further, positively charged residues located within this region are involved in binding, with a lysine triad at positions 55-57 particularly critical. Mutant peptides and protein in which these residues were changed to glutamine retained wild-type structural and functional characteristics by several criteria including circular dichroism spectra, resistance to limited proteolysis, and clathrin binding activity. When expressed in intact cells, mutated alpha subunit showed defective localization to clathrin coated pits; at high expression levels, the appearance of endogenous AP-2 in coated pits was also blocked consistent with a dominant-negative phenotype. These results, together with recent work indicating that phosphoinositides are also critical to ligand-dependent recruitment of arrestin-receptor complexes to coated pits (Gaidarov et al. 1999. EMBO (Eur. Mol. Biol. Organ.) J. 18:871-881), suggest that phosphoinositides play a critical and general role in adaptor incorporation into plasma membrane clathrin-coated pits. PMID- 10459012 TI - VAMP-7 mediates vesicular transport from endosomes to lysosomes. AB - A more complete picture of the molecules that are critical for the organization of membrane compartments is beginning to emerge through the characterization of proteins in the vesicle-associated membrane protein (also called synaptobrevin) family of membrane trafficking proteins. To better understand the mechanisms of membrane trafficking within the endocytic pathway, we generated a series of monoclonal and polyclonal antibodies against the cytoplasmic domain of vesicle associated membrane protein 7 (VAMP-7). The antibodies recognize a 25-kD membrane associated protein in multiple tissues and cell lines. Immunohistochemical analysis reveals colocalization with a marker of late endosomes and lysosomes, lysosome-associated membrane protein 1 (LAMP-1), but not with other membrane markers, including p115 and transferrin receptor. Treatment with nocodozole or brefeldin A does not disrupt the colocalization of VAMP-7 and LAMP-1. Immunoelectron microscopy analysis shows that VAMP-7 is most concentrated in the trans-Golgi network region of the cell as well as late endosomes and transport vesicles that do not contain the mannose-6 phosphate receptor. In streptolysin- O permeabilized cells, antibodies against VAMP-7 inhibit the breakdown of epidermal growth factor but not the recycling of transferrin. These data are consistent with a role for VAMP-7 in the vesicular transport of proteins from the early endosome to the lysosome. PMID- 10459013 TI - Sid2p, a spindle pole body kinase that regulates the onset of cytokinesis. AB - The fission yeast Schizosaccharomyces pombe divides by medial fission through the use of an actomyosin contractile ring. Precisely at the end of anaphase, the ring begins to constrict and the septum forms. Proper coordination of cell division with mitosis is crucial to ensure proper segregation of chromosomes to daughter cells. The Sid2p kinase is one of several proteins that function as part of a novel signaling pathway required for initiation of medial ring constriction and septation. Here, we show that Sid2p is a component of the spindle pole body at all stages of the cell cycle and localizes transiently to the cell division site during medial ring constriction and septation. A medial ring and an intact microtubule cytoskeleton are required for the localization of Sid2p to the division site. We have established an in vitro assay for measuring Sid2p kinase activity, and found that Sid2p kinase activity peaks during medial ring constriction and septation. Both Sid2p localization to the division site and activity depend on the function of all of the other septation initiation genes: cdc7, cdc11, cdc14, sid1, spg1, and sid4. Thus, Sid2p, a component of the spindle pole body, by virtue of its transient localization to the division site, appears to determine the timing of ring constriction and septum delivery in response to activating signals from other Sid gene products. PMID- 10459014 TI - Regulation of APC activity by phosphorylation and regulatory factors. AB - Ubiquitin-dependent proteolysis of Cut2/Pds1 and Cyclin B is required for sister chromatid separation and exit from mitosis, respectively. Anaphase-promoting complex/cyclosome (APC) specifically ubiquitinates Cut2/Pds1 at metaphase anaphase transition, and ubiquitinates Cyclin B in late mitosis and G1 phase. However, the exact regulatory mechanism of substrate-specific activation of mammalian APC with the right timing remains to be elucidated. We found that not only the binding of the activators Cdc20 and Cdh1 and the inhibitor Mad2 to APC, but also the phosphorylation of Cdc20 and Cdh1 by Cdc2-Cyclin B and that of APC by Polo-like kinase and cAMP-dependent protein kinase, regulate APC activity. The cooperation of the phosphorylation/dephosphorylation and the regulatory factors in regulation of APC activity may thus control the precise progression of mitosis. PMID- 10459015 TI - Domains in the 1alpha dynein heavy chain required for inner arm assembly and flagellar motility in Chlamydomonas. AB - Flagellar motility is generated by the activity of multiple dynein motors, but the specific role of each dynein heavy chain (Dhc) is largely unknown, and the mechanism by which the different Dhcs are targeted to their unique locations is also poorly understood. We report here the complete nucleotide sequence of the Chlamydomonas Dhc1 gene and the corresponding deduced amino acid sequence of the 1alpha Dhc of the I1 inner dynein arm. The 1alpha Dhc is similar to other axonemal Dhcs, but two additional phosphate binding motifs (P-loops) have been identified in the NH(2)- and COOH-terminal regions. Because mutations in Dhc1 result in motility defects and loss of the I1 inner arm, a series of Dhc1 transgenes were used to rescue the mutant phenotypes. Motile cotransformants that express either full-length or truncated 1alpha Dhcs were recovered. The truncated 1alpha Dhc fragments lacked the dynein motor domain, but still assembled with the 1beta Dhc and other I1 subunits into partially functional complexes at the correct axoneme location. Analysis of the transformants has identified the site of the 1alpha motor domain in the I1 structure and further revealed the role of the 1alpha Dhc in flagellar motility and phototactic behavior. PMID- 10459016 TI - In vivo, villin is required for Ca(2+)-dependent F-actin disruption in intestinal brush borders. AB - Villin is an actin-binding protein localized in intestinal and kidney brush borders. In vitro, villin has been demonstrated to bundle and sever F-actin in a Ca(2+)-dependent manner. We generated knockout mice to study the role of villin in vivo. In villin-null mice, no noticeable changes were observed in the ultrastructure of the microvilli or in the localization and expression of the actin-binding and membrane proteins of the intestine. Interestingly, the response to elevated intracellular Ca(2+) differed significantly between mutant and normal mice. In wild-type animals, isolated brush borders were disrupted by the addition of Ca(2+), whereas Ca(2+) had no effect in villin-null isolates. Moreover, increase in intracellular Ca(2+) by serosal carbachol or mucosal Ca(2+) ionophore A23187 application abolished the F-actin labeling only in the brush border of wild-type animals. This F-actin disruption was also observed in physiological fasting/refeeding experiments. Oral administration of dextran sulfate sodium, an agent that causes colonic epithelial injury, induced large mucosal lesions resulting in a higher death probability in mice lacking villin, 36 +/- 9.6%, compared with wild-type mice, 70 +/- 8.8%, at day 13. These results suggest that in vivo, villin is not necessary for the bundling of F-actin microfilaments, whereas it is necessary for the reorganization elicited by various signals. We postulate that this property might be involved in cellular plasticity related to cell injury. PMID- 10459017 TI - Integrating the actin and vimentin cytoskeletons. adhesion-dependent formation of fimbrin-vimentin complexes in macrophages. AB - Cells adhere to the substratum through specialized structures that are linked to the actin cytoskeleton. Recent studies report that adhesion also involves the intermediate filament (IF) and microtubule cytoskeletons, although their mechanisms of interaction are unknown. Here we report evidence for a novel adhesion-dependent interaction between components of the actin and IF cytoskeletons. In biochemical fractionation experiments, fimbrin and vimentin coprecipitate from detergent extracts of macrophages using vimentin- or fimbrin specific antisera. Fluorescence microscopy confirms the biochemical association. Both proteins colocalized to podosomes in the earliest stages of cell adhesion and spreading. The complex is also found in filopodia and retraction fibers. After detergent extraction, fimbrin and vimentin staining of podosomes, filopodia, and retraction fibers are lost, confirming that the complex is localized to these structures. A 1:4 stoichiometry of fimbrin binding to vimentin and a low percentage (1%) of the extracted vimentin suggest that fimbrin interacts with a vimentin subunit. A fimbrin-binding site was identified in the NH(2)-terminal domain of vimentin and the vimentin binding site at residues 143 188 in the CH1 domain of fimbrin. Based on these observations, we propose that a fimbrin-vimentin complex may be involved in directing the assembly of the vimentin cytoskeleton at cell adhesion sites. PMID- 10459018 TI - Analysis of CD44-containing lipid rafts: Recruitment of annexin II and stabilization by the actin cytoskeleton. AB - CD44, the major cell surface receptor for hyaluronic acid (HA), was shown to localize to detergent-resistant cholesterol-rich microdomains, called lipid rafts, in fibroblasts and blood cells. Here, we have investigated the molecular environment of CD44 within the plane of the basolateral membrane of polarized mammary epithelial cells. We show that CD44 partitions into lipid rafts that contain annexin II at their cytoplasmic face. Both CD44 and annexin II were released from these lipid rafts by sequestration of plasma membrane cholesterol. Partition of annexin II and CD44 to the same type of lipid rafts was demonstrated by cross-linking experiments in living cells. First, when CD44 was clustered at the cell surface by anti-CD44 antibodies, annexin II was recruited into the cytoplasmic leaflet of CD44 clusters. Second, the formation of intracellular, submembranous annexin II-p11 aggregates caused by expression of a trans-dominant mutant of annexin II resulted in coclustering of CD44. Moreover, a frequent redirection of actin bundles to these clusters was observed. These basolateral CD44/annexin II-lipid raft complexes were stabilized by addition of GTPgammaS or phalloidin in a semipermeabilized and cholesterol-depleted cell system. The low lateral mobility of CD44 in the plasma membrane, as assessed with fluorescent recovery after photobleaching (FRAP), was dependent on the presence of plasma membrane cholesterol and an intact actin cytoskeleton. Disruption of the actin cytoskeleton dramatically increased the fraction of CD44 which could be recovered from the light detergent-insoluble membrane fraction. Taken together, our data indicate that in mammary epithelial cells the vast majority of CD44 interacts with annexin II in lipid rafts in a cholesterol-dependent manner. These CD44 containing lipid microdomains interact with the underlying actin cytoskeleton. PMID- 10459019 TI - Exogenous expression of beta-catenin regulates contact inhibition, anchorage independent growth, anoikis, and radiation-induced cell cycle arrest. AB - beta-Catenin is an important regulator of cell-cell adhesion and embryonic development that associates with and regulates the function of the LEF/TCF family of transcription factors. Mutations of beta-catenin and the tumor suppressor gene, adenomatous polyposis coli, occur in human cancers, but it is not known if, and by what mechanism, increased beta-catenin causes cellular transformation. This study demonstrates that modest overexpression of beta-catenin in a normal epithelial cell results in cellular transformation. These cells form colonies in soft agar, survive in suspension, and continue to proliferate at high cell density and following gamma-irradiation. Endogenous cytoplasmic beta-catenin levels and signaling activity were also found to oscillate during the cell cycle. Taken together, these data demonstrate that beta-catenin functions as an oncogene by promoting the G(1) to S phase transition and protecting cells from suspension induced apoptosis (anoikis). PMID- 10459020 TI - Intracellular movement of green fluorescent protein-tagged phosphatidylinositol 3 kinase in response to growth factor receptor signaling. AB - Phosphatidylinositol 3-kinase (PI 3-kinase) is a lipid kinase which has been implicated in mitogenesis, protein trafficking, inhibition of apoptosis, and integrin and actin functions. Here we show using a green fluorescent protein tagged p85 subunit that phosphatidylinositol 3-kinase is distributed throughout the cytoplasm and is localized to focal adhesion complexes in resting NIH-3T3, A431, and MCF-7 cells. Ligand stimulation of an epidermal growth factor receptor/c-erbB-3 chimera expressed in these cells results in a redistribution of p85 to the cell membrane which is independent of the catalytic activity of the enzyme and the integrity of the actin cytoskeleton. The movement is, however, dependent on the phosphorylation status of the erbB-3 chimera. Using rhodamine labeled epidermal growth factor we show that the phosphatidylinositol 3-kinase and the receptors colocalize in discrete patches on the cell surface. Low concentrations of ligand cause patching only at the periphery of the cells, whereas at high concentrations patches were seen over the whole cell surface. Using green fluorescent protein-tagged fragments of p85 we show that binding to the receptor requires the NH(2)-terminal part of the protein as well as its SH2 domains. PMID- 10459023 TI - Anesthesia for gynecologic laparoscopy. AB - Despite developments in instruments and improvements in surgical and anesthesia techniques, laparoscopy is still associated with complications that may be lethal, including those related to anesthesia. Both anesthesiologist and surgeon must thoroughly understand potential complications of the procedure, including physiologic alterations, principles of anesthetic management and postoperative pain control, and problems related to anesthesia. (J Am Assoc Gynecol Laparosc 6(3):245-258, 1999) PMID- 10459021 TI - Insulin-like growth factor II signaling in neoplastic proliferation is blocked by transgenic expression of the metalloproteinase inhibitor TIMP-1. AB - Insulin-like growth factor (IGF) II is overexpressed in many human cancers and is reactivated by, and crucial for viral oncogene (SV40 T antigen, [TAg])-induced tumorigenesis in several tumor models. Using a double transgenic murine hepatic tumor model, we demonstrate that tissue inhibitor of metalloproteinase 1 (TIMP-1) blocks liver hyperplasia during tumor development, despite TAg-mediated reactivation of IGF-II. Because the activity of IGFs is controlled by IGF-binding proteins (IGFBPs), we investigated whether TIMP-1 overexpression altered the IGFBP status in the transgenic liver. Ligand blotting showed that IGFBP-3 protein levels were increased in TIMP-1-overexpressing double transgenic littermates, whereas IGFBP-3 mRNA levels were not different, suggesting that TIMP-1 affects IGFBP-3 at a posttranscriptional level. IGFBP-3 proteolysis assays demonstrated that IGFBP-3 degradation was lower in TIMP-1-overexpressing livers, and zymography showed that matrix metalloproteinases (MMPs) were present in the liver homogenates and were capable of degrading IGFBP-3. As a consequence of reduced IGFBP-3 proteolysis and elevated IGFBP-3 protein levels, dissociable IGF-II levels were significantly lower in TIMP-1-overexpressing animals. This decrease in bioavailable IGF-II ultimately resulted in diminished IGF-I receptor signaling in vivo as evidenced by diminished receptor kinase activity and decreased tyrosine phosphorylation of the IGF-I receptor downstream effectors, insulin receptor substrate 1 (IRS-1), extracellular signal regulatory kinase (Erk)-1, and Erk-2. Together, these results provide evidence that TIMP-1 inhibits liver hyperplasia, an early event in TAg-mediated tumorigenesis, by reducing the activity of the tumor-inducing mitogen, IGF-II. These data implicate the control of MMP-mediated degradation of IGFBPs as a novel therapy for controlling IGF bioavailability in cancer. PMID- 10459022 TI - Role of transmembrane 4 superfamily (TM4SF) proteins CD9 and CD81 in muscle cell fusion and myotube maintenance. AB - The role of transmembrane 4 superfamily (TM4SF) proteins during muscle cell fusion has not been investigated previously. Here we show that the appearance of TM4SF protein, CD9, and the formation of CD9-beta1 integrin complexes were both regulated in coordination with murine C2C12 myoblast cell differentiation. Also, anti-CD9 and anti-CD81 monoclonal antibodies substantially inhibited and delayed conversion of C2C12 cells to elongated myotubes, without affecting muscle specific protein expression. Studies of the human myoblast-derived RD sarcoma cell line further demonstrated that TM4SF proteins have a role during muscle cell fusion. Ectopic expression of CD9 caused a four- to eightfold increase in RD cell syncytia formation, whereas anti-CD9 and anti-CD81 antibodies markedly delayed RD syncytia formation. Finally, anti-CD9 and anti-CD81 monoclonal antibodies triggered apoptotic degeneration of C2C12 cell myotubes after they were formed. In summary, TM4SF proteins such as CD9 and CD81 appear to promote muscle cell fusion and support myotube maintenance. PMID- 10459025 TI - Histologic studies of the effects of circulating hot saline on the uterus before hysterectomy. AB - STUDY OBJECTIVE: To evaluate a new device for endometrial ablation. DESIGN: (Canadian Task Force classification II-1). SETTING: University-affiliated hospital. PATIENTS: Thirty-two women scheduled for hysterectomy. INTERVENTIONS: Endometrial ablation and hysterectomy. MEASUREMENTS AND MAIN RESULTS: The new device for endometrial ablation was evaluated by studying depth of necrosis after staining for the oxidative enzyme NADH. Uniform endomyometrial necrosis was achieved at a depth of 2 to 4 mm with 90 degrees C saline circulated for 10 minutes. CONCLUSION: The procedure was successful in all patients, and there were no adverse clinical sequelae. (J Am Assoc Gynecol Laparosc 6(3):269-273, 1999) PMID- 10459024 TI - A simple method of coagulating endometrium in patients with therapy-resistant, recurring hypermenorrhea. AB - STUDY OBJECTIVE: To assess the efficacy, safety, and tolerability of hydrothermablation in the treatment of recurrent menorrhagia. DESIGN: Prospective, nonrandomized survey (Canadian Task Force classification II-2). SETTING: Endoscopic center at a university hospital. PATIENTS: Eighteen premenopausal women with recurring menorrhagia resistant to hormone treatment. INTERVENTIONS: Coagulation of the endometrium at 90 degrees C for 10 minutes under hysteroscopic control. MEASUREMENTS AND MAIN RESULTS: During follow-up of least 12 months, nine (50%) of patients had amenorrhea and 17 (94%) had reduction or cessation of pathologic blood flow. One woman underwent hysterectomy because of recurrent dysmenorrhea. CONCLUSION: In this study, hydrothermablation was a safe and effective method of treating recurrent menorrhagia. It does not require extensive training and may help avoid hysterectomy. (J Am Assoc Gynecol Laparosc 6(3):265-268, 1999) PMID- 10459026 TI - Evaluation of Hydro ThermAblator for endometrial destruction in patients with menorrhagia. AB - OBJECTIVE: To describe endometrial destruction with the Hydro ThermAblator. DESIGN: (Canadian Task Force classification II-1). SETTING: University-affiliated hospital. PATIENTS: Twenty-six women with uncontrolled menorrhagia. INTERVENTION: Endometrial ablation with saline heated to 90 degrees C and circulated in the uterine cavity for 10 minutes under hysteroscopic control. MEASUREMENTS AND MAIN RESULTS: All patients tolerated the procedure well, all but one was satisfied with the results, and there were no significant side effects or complications. Seventy-seven percent of women were either amenorrheic or hypomenorrheic after 6 months, and 88% were amenorrheic or hypomenorrheic after 1 year, including one patient who was treated a second time; 87.5% were amenorrheic or hypomenorrheic at 18 months. One woman had persistent menorrhagia and was treated by hysterectomy. CONCLUSION: In this study, endometrial ablation was performed successfully with the Hydro ThermAblator in 25 of 26 women. (J Am Assoc Gynecol Laparosc 6(3):275-278, 1999) PMID- 10459027 TI - Selective uterine artery embolization as primary treatment for symptomatic leiomyomata uteri. AB - STUDY OBJECTIVE: To analyze initial experience with uterine artery embolization for treatment of symptomatic leiomyomata. DESIGN: Prospective, longitudinal study (Canadian Task Force classification II-2). SETTING: Private practice, university affiliated hospital. PATIENTS: Three hundred five women (age 26-52 yrs). INTERVENTIONS: Uterine artery embolization, performed over 2 years by a single radiologist working in collaboration with a single gynecology practice. MEASUREMENTS AND MAIN RESULTS: Embolization was technically successful in 96% of patients. No major complications occurred. Average reduction in uterine volume was 48%. Control of menorrhagia was reported by 86% of patients at 3 months, 85% at 6 months, and 92% at 12 months after the procedure. Bulk symptoms were satisfactorily controlled in 64% of patients at 3 months, 77% at 6 months, and 92% at 12 months. Six women subsequently underwent hysterectomy and five had myomectomy. CONCLUSION: Uterine artery embolization appears to be a highly effective treatment for symptomatic uterine leiomyomata. Its impact on fertility and pregnancy remain to be investigated fully. (J Am Assoc Gynecol Laparosc 6(3):279-284, 1999) PMID- 10459028 TI - A randomized, prospective study of endometrial resection to prevent recurrent endometrial polyps in women with breast cancer receiving tamoxifen. AB - STUDY OBJECTIVE: To assess the role of endometrial resection in preventing recurrence of tamoxifen-associated endometrial polyps in women with breast cancer. DESIGN: Randomized, prospective study (Canadian Task Force classification I). SETTING: Tertiary university-affiliated medical center. PATIENTS: Twenty consecutive women (age range 43-61 yrs). INTERVENTIONS: Hysteroscopic removal of tamoxifen-associated endometrial polyps with or without simultaneous resection of the endometrium. MEASUREMENTS AND MAIN RESULTS: Patients were randomized to undergo (10 women) or not undergo (10) concomitant endometrial resection. They were followed for at least 18 months (range 18-24 mo), including transvaginal ultrasonography every 6 months and hysteroscopy when endometrial irregularity was noted. The main outcome variable was recurrence of endometrial polyps; occurrence of uterine bleeding was also noted. In women who underwent endometrial resection, only one had a 1 x 1-cm endometrial polyp diagnosed and removed during follow-up. Seven women remained amenorrheic, and three experienced spotting for a few days every month. In the control group, six women had recurrent endometrial polyps requiring hysteroscopic removal (two-tail Fisher's exact test p <0.06). CONCLUSION: Recurrence of endometrial polyps, one of the most common problems in patients with breast cancer receiving long-term treatment with tamoxifen, may be reduced by performing endometrial resection at the time of hysteroscopic removal of polyps. The possible risk of occult endometrial cancer is yet to be determined. (J Am Assoc Gynecol Laparosc 6(3):285-288, 1999) PMID- 10459029 TI - Measurement of CO(2) hypothermia during laparoscopy and pelviscopy: how cold it gets and how to prevent it. AB - STUDY OBJECTIVE: To evaluate intraabdominal CO(2) temperature during a variety of standard operative laparoscopy procedures with different insufflators (BEI Medical, Snowden & Pencer, Storz Laparoflator, Storz Endoflator, Wolf) and devices to maintain body temperature (Bair Hugger, fluid warmer, Blanketrol blankets). DESIGN: Prospective, nonrandomized study (Canadian Task Force classification II-1). SETTING: Community hospital in rural Alabama. PATIENTS: Sixty-two consecutive patients (53 women, 9 men; average age 56.8 yrs, range 21 94 yrs). INTERVENTIONS: Patients underwent standard laparoscopic and pelviscopic procedures during which intraoperative temperature changes in the insufflation system, abdomen, and rectum were measured. MEASUREMENTS AND MAIN RESULTS: Carbon dioxide was at room temperature in the insufflation hose ( approximately 23 degrees C). During insufflation, intraabdominal gas temperature decreased to as much as 27.7 degrees C (average 32.7 degrees C) depending on length of operation (23 min-5 hrs 8 min), amount of gas used (12.8-801 L), gas flow (up to 20 L/min), and leakage rate. Preoperative and postoperative temperature comparisons showed no decline in rectal temperature (average +0.18 degrees C) because warming equipment was sufficient. CONCLUSION: The decrease in intraoperative intraabdominal gas temperature is remarkable and can potentially harm the patient. It can be limited by restricting gas flow and leakage. In operations longer than 1 hour, substantial core body temperature drop should be prevented with appropriate heating and hydration devices. An insufflator with internal gas heating (Snowden & Pencer) had no significant clinical effect. (J Am Assoc Gynecol Laparosc 6(3):289-295, 1999) PMID- 10459030 TI - Operative laparoscopic treatment of ovarian retention syndrome. AB - STUDY OBJECTIVE: To evaluate the ability to treat ovarian retention syndrome (ORS) by operative laparoscopy. DESIGN METHODS: Retrospective observational analysis (Canadian Task Force classification II-2). SETTING: Pelvic pain referral practice in an university-affiliated community hospital. PATIENTS: Thirty consecutive women with ORS. INTERVENTION: Operative laparoscopy. MEASUREMENTS AND MAIN RESULTS: Salpingo-oophorectomy or oophorectomy was completed laparoscopically in 26 women; conversion to laparotomy was necessary in 4 (13.3%). Mean operating time was 159 +/- 63 minutes, mean blood loss was 119 +/- 152 ml, and mean hospital stay was 1.1 +/- 1.4 days. Complications occurred in four patients (13. 3%); in two (6.6%) they were intraoperative. The mean time interval between hysterectomy and symptoms of ORS was 6.4 +/- 4.6 years. Mean preoperative duration of pain was 40.11 +/- 41.3 months. Of 27 patients with adequate follow-up, 13 (48%) were pain free postoperatively with a mean follow-up of 12.9 +/- 8.2 months. Fourteen women (52%) had recurrent pain with mean time to recurrence of 8.8 +/- 10 months. Mean visual analog pain scores were 7.5 +/- 2. 4 preoperatively and 2.3 +/- 3.1 postoperatively (p <0.001). Endometriosis at time of surgery was associated with a significantly higher risk of recurrent pelvic pain (relative risk = 2.3, 95% confidence intervals 1.1, 5.1). Ovarian preservation was significantly related to recurrence of pain (RR = 2.6, 95% CI 1.52, 4.53) and risk of repeat surgery (RR = 4.4, 95% CI 1.69, 11.33). CONCLUSIONS: Ovarian retention syndrome can usually be treated by laparoscopy. Removal of both ovaries, if present, may be necessary to prevent recurrent pain. In our series, 48% of women experienced prolonged relief. (J Am Assoc Gynecol Laparosc 6(3):297-302, 1999) PMID- 10459031 TI - Carcinoid tumors of the appendix detected at laparoscopy for gynecologic indications. AB - STUDY OBJECTIVE: To evaluate the significance of the abnormal-appearing appendix during gynecologic laparoscopy. DESIGN: Retrospective study (Canadian Task Force classification II-2). SETTING: University-affiliated hospital. PATIENTS: One hundred sixty-six women. INTERVENTIONS: Laparoscopic appendectomy. MEASUREMENTS AND MAIN RESULTS: Five of 166 appendixes removed for abnormal appearance contained a carcinoid tumor. At least one patient was a candidate for additional resection. CONCLUSION: It is important for gynecologists to be aware of the possibility of a carcinoid tumor of the appendix, particularly if an abnormality is seen at the time of surgery. (J Am Assoc Gynecol Laparosc 6(3):303-306, 1999) PMID- 10459032 TI - Laparoscopy in morbidly obese patients. AB - STUDY OBJECTIVE: To assess the safety and efficacy of different insufflation methods in morbidly obese women undergoing laparoscopy. DESIGN: Retrospective analysis of 13 years' experience (Canadian Task Force classification II-2). SETTING: University-affiliated hospital. PATIENTS: One hundred thirty-eight morbidly obese women (weight >250 lbs, body mass index >36). The heaviest patient weighed 400 lbs and had a body mass index of 66. INTERVENTION: Laparoscopic tubal sterilizations and diagnostic laparoscopies performed on an outpatient basis by residents under faculty supervision. MEASUREMENTS AND MAIN RESULTS: Of 138 patients, 36 underwent standard transumbilical insufflation with 5 failures, 83 had transuterine insufflation with 3 failures, 12 had subcostal insufflation with 1 failure, and 7 had open laparoscopy with 2 failures. CONCLUSION: The insufflation failure rate was significantly high for transumbilical insufflation and open laparoscopy, and not for transuterine or subcostal insufflation. Morbid obesity was not a contraindication to laparoscopy. (J Am Assoc Gynecol Laparosc 6(3):307-312, 1999) PMID- 10459033 TI - Laparoscopic management of selected adnexal masses. AB - STUDY OBJECTIVE: To investigate the significance, safety, and intraoperative and immediate postoperative outcomes of laparoscopic management of adnexal masses thought to be at low risk for malignancy. DESIGN: Prospective cohort study (Canadian Task Force classification II-2). SETTING: Tertiary-care teaching hospital. PATIENTS: Two hundred twenty women undergoing laparoscopic surgery for adnexal masses. INTERVENTIONS: Laparoscopic treatment including cystectomy, oophorectomy, adnexectomy, and peritoneal cytology, and, if necessary, frozen sections. A histologic diagnosis was obtained in every patient. MEASUREMENTS AND MAIN RESULTS: Only one ovarian cancer and one borderline ovarian tumor were diagnosed by histologic examination, and both were managed by laparotomy. The remaining 218 patients had laparoscopy for benign adnexal masses. CONCLUSION: Operative laparoscopy with the finding of incidental ovarian malignancy is rare, as shown by pathologic examination. With appropriate preoperative evaluation, laparoscopic surgery is technically feasible, safe, and advantageous, with minimal morbidity, and should replace laparotomy in the management of most adnexal masses. (J Am Assoc Gynecol Laparosc 6(3):313-316, 1999) PMID- 10459034 TI - Complications of gynecologic laparoscopy. AB - STUDY OBJECTIVE: To examine the frequency of gynecologic laparoscopy complications from January 1996 to June 1996. DESIGN: Prospective review (Canadian Task Force classification II-1). SETTING: Tertiary center teaching hospital with 607 beds. PATIENTS: Two hundred thirty-four patients (age 15-70 yrs). INTERVENTIONS: Gynecologic laparoscopy. MEASUREMENTS AND MAIN RESULTS: The overall number of complications was 23 (9.8%). They were principally infection, extraperitoneal insufflation, and injury to blood vessels. Complication rates were examined by service (private, clinic, staff) and by type of laparoscopy (operative, diagnostic, laparoscopic-assisted vaginal hysterectomy). Of 11 (47.8%) complications that occurred during laparoscopic-assisted vaginal hysterectomy, 5 involved postoperative infection. CONCLUSION: The complication rate in this review was similar to that published in the literature, with the exception of ileus and infection, which occurred at higher rates in our institution. (J Am Assoc Gynecol Laparosc 6(3):317-321, 1999) PMID- 10459035 TI - Laparoscopy using the left upper quadrant approach. AB - STUDY OBJECTIVE: To describe a technique of laparoscopy and multiport operative pelviscopy using left upper quadrant primary port insertion. DESIGN: Retrospective evaluation (Canadian Task Force classification II-2). SETTING: Private gynecologic oncology practice. PATIENTS: Ninety women with a history of gynecologic cancer and at least one laparotomy. INTERVENTIONS: Laparoscopy and operative pelviscopy. MEASUREMENTS AND MAIN RESULTS: The procedure was performed using the left upper quadrant as a single entry site for the Veress needle and primary laparoscopy port. In 88 women it was performed without complication. One woman experienced transverse colon injury from primary port insertion, which was repaired immediately by laparotomy. A second patient had a rectosigmoid injury that required temporary colostomy at laparotomy. CONCLUSION: Operative laparoscopy using the left upper quadrant approach seems to be safe in patients with advanced gynecologic malignancy. (J Am Assoc Gynecol Laparosc 6(3):323-325, 1999) PMID- 10459036 TI - Laparoscopic creation of a neovagina in women with pelvic kidney. AB - Laparoscopic modification of Vecchietti's technique for creating a neovagina was carried out in four women with Rokitansky syndrome and renal anomalies. All four patients had pelvic kidney, and in two it was associated with contralateral renal agenesis. The procedure was specifically modified to reduce the risks inherent in the most difficult step, passing the thread-bearing cutting needle from the abdominal wall to the retrohymenal fossa, through the vesicorectal space. In all patients this was done successfully on the side contralateral to the pelvic kidney or in the hemipelvis in which the ureter was absent. Our experience proves that Vecchietti's technique for creating a neovagina, performed by laparoscopy, is feasible in patients with pelvic kidney, and laparotomy is unnecessary. (J Am Assoc Gynecol Laparosc 6(3):327-329, 1999) PMID- 10459037 TI - A new bipolar system for performing operative hysteroscopy in normal saline. AB - We evaluated the efficacy and safety of the VersaPoint bipolar vaporization system during hysteroscopic treatment of 10 women with symptomatic submucous leiomyomata, endometrial polyps, uterine septa, and synechiae in a prospective, uncontrolled pilot study at two Canadian university-affiliated teaching hospitals. Patients were two women with recurrent pregnancy loss associated with a diagnosis of uterine septa, one with infertility, one with recurrent pregnancy loss and synechiae, and six with menorrhagia associated with either leiomyomata or endometrial polyps. Electrodes were inserted through a 5F operating channel of a 15F cystoscope, and a 17 or 21F hysteroscope. Three electrodes were used: ball, twizzle, and spring. Power settings ranged from 50 W (desiccation mode) to 200 W (vapor cut mode). Normal saline was used as the distention medium in all cases. Either general anesthesia or intravenous sedation with paracervical block was used. There were no major complications such as uterine perforation, excessive bleeding, fluid overload, or thermal injury. The amount of normal saline used varied considerably from 0.5 to 20 L. The maximum amount of saline absorbed was 900 ml in a case involving resection of 4.5- and 2.0-cm leiomyomata that lasted 115 minutes. Mild cramping, vaginal bleeding, and discharge in the first 2 to 3 days were relieved by nonsteroidal antiinflammatory agents. No patients were readmitted up to 6 weeks after the procedure. Preliminary results of this pilot study support the safety of the VersaPoint bipolar vaporization system, although its long-term efficacy remains to be determined. It appears to be well tolerated by some women using conscious sedation and paracervical block. Therefore, it can potentially be used in an office setting as well as in symptomatic patients who are at risk from a general anesthetic. (J Am Assoc Gynecol Laparosc 6(3):331-336, 1999) PMID- 10459038 TI - Chain removal of myomata by colpotomy. AB - Multiple myomectomy was performed in 67 women between June 1996 and July 1998. In almost every case, myomas were sutured one to another with a single stitch of 1 silk, forming a chain, and taken out through the vaginal cul-de-sac. We believe this method reduces the risk of losing myomas inside the peritoneal cavity and shortens operating time. (J Am Assoc Gynecol Laparosc 6(3):337-338, 1999) PMID- 10459039 TI - Laparoscopic treatment of vesicouterine fistula. AB - Vesicouterine fistula are rare disorders that are most commonly repaired by laparotomy. A specific type of vesicouterine fistula, vesicocervical fistula, was repaired successfully by laparoscopy. This procedure was associated with minimal blood loss and morbidity, and the patient's hospital stay was less than 24 hours. (J Am Assoc Gynecol Laparosc 6(3):339-341, 1999) PMID- 10459040 TI - Small bowel obstruction in a peritoneal defect after laparoscopic Burch procedure. AB - With expansion of laparoscopic surgery, the gynecologic surgeon faces a new set of complications not encountered in open surgery and completely related to the laparoscopic approach. A rare complication occurred after laparoscopic Burch colposuspension performed in a patient with genuine stress urinary incontinence. Although the fascial incision at the right lower quadrant cannula insertion site was closed, a loop of small bowel herniated through the nonclosed peritoneal defect and caused intestinal obstruction 9 days after surgery. For cannula insertion sites 10-mm and greater, we recommend suturing peritoneal defects at the time of fascial closure. (J Am Assoc Gynecol Laparosc 6(3):343-345, 1999) PMID- 10459041 TI - Pregnancy and laparoscopic surgery. AB - We reviewed the English literature regarding laparoscopic surgery during pregnancy and found that of 518 reported procedures, the most common was cholecystectomy (45%), followed by adnexal surgery (34%), appendectomy (15%), and other operations (6%). We add six cases to this list; three cholecystectomies, an adnexal procedure, and two for abdominal pain. Thirty-three percent were performed in the first trimester, 56% in the second, and 11% in the third trimester. This review demonstrates a definite trend, indicating that laparoscopy in pregnancy appears to be safe when performed by experienced practitioners. (J Am Assoc Gynecol Laparosc 6(3):347-351, 1999) PMID- 10459042 TI - Hsp47-dependent and -independent intracellular trafficking of type I collagen in corneal endothelial cells. AB - PURPOSE: Type I collagen is post-translationally regulated in corneal endothelial cells (CEC): CEC synthesize procollagen I and degrade it intracellularly. We investigated whether there is a Hsp47-independent pathway during intracellular trafficking of procollagen I. METHODS: Specific inhibitors were used to block intracellular transport of procollagen I and Hsp47. Immunocytochemical analysis was performed to determine the intracellular localization of the proteins of interest. RESULTS: When cells were treated with alpha,alpha'-dipyridyl, this specific inhibitor for collagen promoted retention in the endoplasmic reticulum (ER) of some of the underhydroxylated procollagen I, which was colocalized with Hsp47 in CEC. At the same time, another fraction of the alpha,alpha'-dipyridyl induced underhydroxylated procollagen I was not located in the ER. When CEC were treated with brefeldin A, procollagen I and Hsp47 demonstrated a high degree of colocalization at the ER, whereas the inhibitor had less of an effect on the compartmentalization of procollagen I and prolyl 4-hydroxylase. When CEC were treated with either monensin or bafilomycin A1, procollagen I and Hsp47 were not colocalized: procollagen I was mostly localized at the Golgi area, while Hsp47 predominantly showed ER distribution. When colocalization of procollagen I and prolyl 4-hydroxylase was examined, a major population of procollagen I was not colocalized with prolyl 4-hydroxylase in the ER. CONCLUSIONS: These results indicate that some procollagen I and Hsp47 travel together from the ER to the cis Golgi compartment and that a major population of procollagen I that may not be properly hydroxylated may be destroyed intracellularly via the Hsp47-independent pathway in CEC. PMID- 10459043 TI - FGF-2 facilitates binding of SH3 domain of PLC-gamma1 to vinculin and SH2 domains to FGF receptor in corneal endothelial cells. AB - PURPOSE: To determine the cellular localization of the Src homology (SH)2 and SH3 domains of PLC-gamma1 and their cytoplasmic binding partners, living corneal endothelial cells were microinjected with the fusion proteins containing SH domains. METHODS: Fusion proteins were prepared from plasmid vectors, and the fusion proteins containing SH2-SH2 [(SH2)2], SH2-SH2-SH3 [(SH2)2-SH3] or SH3 were isolated using affinity chromatography. Following microinjection, immunolocalization was analyzed using confocal laser microscope. RESULTS: Microinjected SH domains were targeted to the subcellular location following stimulation with FGF-2: the SH3 domain appeared to be targeted to cytoskeleton; the (SH2)2 domain showed a dual localization in cytoplasm and plasma membrane; the (SH2)2-SH3 domain was predominantly localized at membrane and perinuclear sites. In the absence of stimulation by FGF-2, the microinjected fusion proteins remained at the injection sites. When cytoplasmic binding partners were determined by double-staining, the SH3 domain demonstrated colocalization with vinculin: the staining profile of the SH3 domain was identical to that of vinculin, which demonstrates characteristic punctated profiles. The punctated staining of SH3 disappears toward the basal membrane, while that of vinculin remains in all confocal optical sections. On the other hand, some fraction of the (SH2)2 domain was colocalized with FGF receptor at the membrane site. When PLC gamma1 and F-actin were double-stained, the endogenous PLC-gamma1 demonstrated a diffuse cytoplasmic staining and/or perinuclear staining, while phalloidin staining demonstrated that all cells have filamentous cytoplasmic distribution of F-actin. CONCLUSIONS: These findings indicate that the SH3 domain directs PLC gamma1 to bind to vinculin and that the SH2 domains may mediate the binding of PLC-gamma1 to receptor tyrosine kinase. Furthermore, they suggest that phosphorylation is not required for targeting of PLC-gamma1 to membrane or cytoskeleton sites. PMID- 10459044 TI - A trabecular meshwork glucocorticoid response (TIGR) gene mutation affects translocational processing. AB - PURPOSE: To examine possible effects of the E323K mutation in the trabecular meshwork glucocorticoid response (TIGR) gene (also known as myocilin [MYOC]), using assays of translocational processing through the endoplasmic reticulum (ER). The E323K mutation was of particular interest, since the mutation shows a strong association with early onset open-angle glaucoma, but has a minimal predicted effect on protein structure. METHODS: Normal and mutant TIGR cDNA constructs were used to generate protein products in the presence of endoplasmic reticulum (ER) membranes, using an assay previously developed to detect alterations in the ER translocation function. "Paused" regions for potential protein modifications were defined by proteinase K (PK) sensitivity in the presence of ER membranes, with the ability to restart translocation when treated with EDTA. The effects of the E323K mutation were evaluated, as well as mutations located on either side of E323K (G246R, G364V, P370L) as the other mutations had substantial predicted structural changes in addition to clear disease associations. RESULTS: The native TIGR molecule was observed to have a paused region that corresponds to the region of highest olfactomedin (OLF) homology. The E323K mutation, located near the beginning of this region, dramatically altered the normal pattern of nascent proteins observed in the translocational pausing assay. A prominent band appeared with the E323K mutation, which could represent a new product or a marked enhancement of a faint band normally seen, approximately 3 kDa higher than the major paused band. The other TIGR mutants examined did not show this effect. CONCLUSIONS: The major translocational pause that starts near the beginning of the region of high OLF homology may help to explain the high frequency of glaucoma-associated mutations in this area. The observed effect of the E323K mutation on the products of translocational processing suggests a delay in the normal pausing process of TIGR biogenesis. This delay points to a potentially distinct pathogenic mechanism for E323K as compared with the other TIGR mutations so far evaluated. PMID- 10459045 TI - Epithelial tumors of the lacrimal glands: a clinicopathologic study. AB - We report the clinicopathologic features of epithelial tumors of the lacrimal gland apparatus, which are rare and therefore represent a major challenge for diagnosis and treatment. Histologic material from 22 lesions was studied by light microscopy, histochemistry, and immunohistochemistry. A comparison with major and minor salivary gland tumors was performed to analyze the relative distribution of these tumors and to establish whether salivary glands and lacrimal gland tumors are similar or different in their pathologic appearance and clinical behavior. There were three benign pleomorphic adenomas and 19 malignant tumors. The gender distribution was equal. The ages of the patients ranged from 10 to 73 years (mean age, 46 years). Among the malignant tumors, adenoid cystic carcinoma was the most common (nine cases), followed by mucoepidermoid carcinoma (three cases). There were two cases each of malignant mixed tumor and adenocarcinoma. All mucoepidermoid carcinomas and the adenocarcinomas were histologically high grade. There also was one case each of salivary duct carcinoma, spindle cell carcinoma, and oncocytic adenocarcinoma. Of 14 patients in whom clinical follow-up was available, seven had distant metastases and four died of their disease. The only case occurring in a child was an adenoid cystic carcinoma that recurred locally after 14 years. The clinical and pathologic features of lacrimal gland tumors resemble those lesions that arise in the intraoral minor salivary glands. The greater relative proportion of malignant cases in this series probably reflects a selection bias. PMID- 10459046 TI - Histopathologic findings in 37 cases of functional hemispherectomy. AB - Hemispherectomy procedures are performed in patients for whom focal cortical resection would be predicted to produce a significant reduction in seizures. The functional hemispherectomy procedure consists of disconnecting the hemispheres while attempting, in some cases, to preserve parenchyma. This study retrospectively reviews the histopathologic findings in 37 cases of functional hemispherectomy performed between 1990 and 1998 at a major epilepsy center. Procedures were performed in 20 males and 17 females who ranged in age from 3 months to 37 years (mean age, 9.6 years). In all but two cases, more than half or all the material submitted for pathologic testing was examined histologically. Cortical dysplasias or hemimegalencephaly were identified in 14 patients. The most common patterns of dysplasia observed included architectural disorganization (n = 13), increased molecular layer neurons (n = 11), and neuronal cytomegaly (n = 11). One patient was known to have epidermal nevus syndrome. Six patients had Sturge-Weber syndrome. Remote infarct/ischemic damage was identified as the etiology of seizures in six patients; four of these patients had mild associated secondary cortical architectural abnormalities. Three patients demonstrated pathology consistent with Rasmussen's encephalitis; one additional patient had chronic encephalitis changes, not otherwise specified. In two cases, changes consistent with hippocampal sclerosis were identified; additionally, hippocampal neuronal loss and gliosis was focally identified in three patients. Most of these patients had coexistent cortical dysplasia or radiographic evidence of remote infarct. One specimen demonstrated areas of infarct following resection of an arteriovenous malformation. In two specimens, significant histopathologic findings were not identified; both of these patients had radiographic evidence of remote infarct. The spectrum of pathologic conditions that may be encountered in the setting of a functional hemispherectomy is varied and in this study most frequently included cortical dysplasia, Sturge-Weber syndrome remote infarct, and Rasmussen's encephalitis. PMID- 10459047 TI - Malignant Sertoli cell tumor of the ovary metastatic to the lung mimicking neuroendocrine carcinoma: report of a case. AB - A Sertoli cell carcinoma of the ovary with lung metastases mimicking neuroendocrine carcinoma is presented. Lung metastases frequently occur. Primary and secondary tumors may exhibit similar growth patterns and differentiating primary from secondary tumors may be troublesome. This process may be more difficult when metastases occur from a tumor in which metastases are uncommon and morphologically resemble only a small portion of the primary tumor. We report the case of a 52-year-old woman who underwent resection of a 4,550-g Sertoli cell tumor of the ovary. Histologically, in addition to the characteristic tubular pattern of growth, 5% of the tumor consisted of poorly differentiated areas with tumor cells in sheets, a high mitotic rate, and areas of necrosis. Eleven months after this surgery she presented at a different institution with multiple pulmonary nodules. Microscopic examination of a subsequently resected lung nodule showed histologic findings similar to those of the poorly differentiated areas of the ovarian tumor and initial immunohistochemical studies showed positive staining for cytokeratin, neuron-specific enolase, and focal positivity for synaptophysin. Without knowledge of the ovarian tumor the lung lesion was interpreted as large-cell neuroendocrine carcinoma. On review of the clinical history and comparison with the previous surgical material, however, both tumors showed similar light microscopy and immunohistochemical reactivity, and a final diagnosis of metastatic Sertoli cell tumor was made. Immunohistochemical staining for inhibin revealed weak positivity in the poorly differentiated areas of the ovarian tumor but not in the lung metastasis. This is one of the rare reports of ovarian Sertoli cell tumor metastasizing to the lungs and it emphasizes the importance of complete clinical histories, ancillary studies, appropriate sampling, and review of archival material in such unusual cases. PMID- 10459048 TI - Signet-ring cell variant of small lymphocytic lymphoma with a prominent sinusoidal pattern. AB - Signet-ring cell lymphoma is a rare morphologic variant of non-Hodgkin's lymphoma characterized by neoplastic lymphoid cells with cytoplasmic vacuoles or eosinophilic globules that impart a signet-ring cell morphology. Although most cases are variants of follicular center B-cell lymphomas, this pattern also can be seen in T-cell lymphomas. An indolent clinical course and prolonged survival have characterized the majority of published cases. We document the case of a 62 year-old African-American woman with diffuse small lymphocytic signet-ring lymphoma having a predominant sinusoidal growth pattern, which, to our knowledge, has not been previously reported. The prominent sinusoidal pattern of signet-ring lymphocytes contributes to its confusion with metastatic signet-ring cell adenocarcinoma. The correct diagnosis is greatly facilitated by the use of appropriate immunohistochemical stains for lymphoid markers. PMID- 10459049 TI - Myxomatous soft tissue tumors: correlation of cytopathology and histopathology. AB - A small, uncommon group of soft tissue tumors are distinguished by their unique and consistent ability to produce an overwhelming abundance of myxoid ground substance along with the proliferating cells that constitute the tumor. Grossly, all these neoplasms have a variable gelatinous quality. Because of the voluminous stroma, most of these tumors have some findings that overlap with one another. Nonetheless, each tumor has a composite set of morphologic, immunophenotypic, ultrastructural, and genotypic features exclusive to itself. Because soft tissue masses are not a frequent site of fine-needle aspiration, the cytopathology of this set of tumors is vastly unappreciated, both in the literature and in day-to day practice. The aim of this review is to detail the salient light microscopic findings of this group of six major myxoid soft tissue tumors, to correlate the cytopathology (particularly as obtained using the fine-needle aspiration biopsy technique) with its histopathologic counterparts, and to discuss the limitations of both cytologic and histologic methods. This cytohistopathologic correlation should assist the reader in the diagnosis of myxoid tumors of soft tissue. PMID- 10459050 TI - Aggressive (malignant) epithelial odontogenic ghost cell tumor. AB - A 57-year-old man had an expanding cystic lesion of the anterior maxilla that demonstrated destruction and disruption of local structures. A cystic odontogenic neoplasm as well as various forms of odontogenic carcinoma was suspected. Incisional biopsy and microscopic examination revealed an aggressive epithelial odontogenic tumor with histologic features suggesting malignant transformation from an odontogenic cyst. The patient continues to refuse further treatment despite remaining tumor. This case further documents the malignant potential of a calcifying odontogenic cyst and the existence of at least an aggressive or possibly malignant form of epithelial odontogenic ghost cell tumor. To date, metastatic work-up has not been performed. PMID- 10459051 TI - Uncommon morphologic patterns of lobular neoplasia. AB - Discussions of the morphology of lobular neoplasia (atypical lobular hyperplasia and lobular carcinoma in-situ) usually emphasize the commonplace characteristics. Occasional cases deviate from the expected appearance and thereby cause diagnostic confusion. Unusual cytologic alterations include the formation of signet-ring cells and the presence of nuclear pleomorphism. The growth of lobular neoplasia in breast tissue altered by atrophy, sclerosing adenosis, collagenous spherulosis, and benign tumors gives rise to unexpected patterns that one might mistake for normal tissue, invasive carcinoma, or ductal carcinoma in situ. Solid involvement of ducts by lobular neoplasia and uncommon forms of "pagetoid" growth both resemble the appearance of atypical ductal hyperplasia or ductal carcinoma in situ. The authors illustrate these unusual cytologic and architectural findings and discuss features that permit proper diagnosis and interpretation. PMID- 10459052 TI - Henry L. Jaffe, MD (1896-1979), a pioneering authority on bone diseases: reflections and an appreciation. AB - The almost forgotton American pioneer in the history of bone pathology Henry L. Jaffe, MD died 20 years ago. He was instrumental in first describing many pathologic osseous conditions, both neoplastic and non-tumorous. His scientific achievement qualifies Dr Jaffe as a true giant of bont pathology. PMID- 10459053 TI - An application to join a prestigious slide club PMID- 10459054 TI - Preface PMID- 10459055 TI - Laparoscopic anatomy of the pelvis. AB - The laparoscopic anatomy of the pelvis is reviewed. Both male and female anatomy are detailed, and special emphasis is placed on avoiding anatomic complications of laparoscopic pelvic surgery. PMID- 10459056 TI - Microlaparoscopy. AB - Microlaparoscopy is defined as using instruments with an outer sheath of less than 2 mm; as such, it represents the leading edge of fiberoptic and instrument design technology. Although still in its infancy, it has been proposed as the new standard instrument for abdominal entry and for the performance of some diagnostic and therapeutic procedures. It is already the instrument of choice for performing conscious laparoscopic procedures. The small size of the instrument makes it versatile, but it is important that they are used appropriately. In this way, microlaparoscopy offers significant advantages over conventional laparoscopy for surgeon and patient alike. PMID- 10459057 TI - Laparoscopic surgery of the fallopian tubes and ovaries. AB - Operative laparoscopy can be used for many surgical procedures on the fallopian tube and ovary. These include: (1) tubal sterilization; (2) salpingectomy and salpingostomy for tubal pregnancy; (3) fimbrioplasty, salpingoneostomy, and linear salpingostomy for tubal obstruction and infertility; (4) microsurgical tubal reanastomosis for reversal of tubal sterilization; (5) oophorectomy, cystectomy, cyst drainage and fulguration, and excision of ovarian tumors; (6) wedge resection and ovarian drilling for polycystic ovaries; and (7) fulguration and laser vaporization for endometriosis. Many of these procedures are conservative and involve reconstruction of the tube and ovaries to preserve fertility. Microsurgical techniques are incorporated into such fertility sparing or enhancing procedures. Comparison of similar surgical procedures on the tube and ovaries indicates better or similar surgical outcome when done through the laparoscope rather than laparotomy; less blood loss, faster recovery, and cheaper cost are the hallmarks when the procedure is done by laparoscopy. With further improvement and expansion in laparoscopy equipment, it can be expected that more surgical procedures on the adnexa can be undertaken safely and effectively. PMID- 10459058 TI - Laparoscopic management of ectopic pregnancy. AB - The incidence of ectopic pregnancy has been rising over the last 20 years. The cause is multifactorial. The technical advancement in the field of minimal access surgery has greatly enhanced the possibility of both diagnosing and treating the condition effectively. The management of ectopic pregnancy can be expectant and surgical; the latter can be by open or laparoscopic methods. Laparoscopic surgery is usually performed when the patient is haemodynamically stable, the hCG is <6000 IU/L, the history is suggestive of minimal pelvic adhesions and when the pregnancy is confined within the tube. This procedure is, however, vastly dependent on the experience and expertise of the surgeon and the equipment facilities available. There are various means of treating ectopic pregnancy by laparoscopy. This includes laparoscopic salpingectomy, salpingotomy, and direct injection of cytotoxic agents. This article will present an overview to ascertain the effectiveness of minimal access surgery in managing ectopic pregnancy. There seems little doubt that laparoscopic management should be the first line of treatment. The way to realize its full potential is by formulating clear guidelines regarding the indication and implementation of laparoscopic surgery and emphasising the role of a fully structured training program to achieve the goals. PMID- 10459059 TI - Laparoscopic myomectomy. AB - Fibroids are common tumors that present with a variety of symptoms for whom medical treatment is generally unsatisfactory. Laparoscopic myomectomy was described nearly 20 years ago. Since then several hundred procedures have been described in the literature. With increasing experience it has become apparent that the technique is both technically demanding and time consuming. It is also not clear whether laparoscopic surgical outcome is improved compared with laparotomy. At present these factors limit the application of laparoscopic myomectomy. PMID- 10459060 TI - Laparoscopic hysterectomy. AB - The first laparoscopic hysterectomy using only laparoscopic techniques was performed around 10 years ago by Harry Reich. Such total laparoscopic hysterectomy (TLH) approach is surgically elegant but technically difficult. A number of alternative laparoscopic techniques to perform all or some of the hysterectomy have since been introduced to simplify the surgical technique but retain the major advantages of the approach, which is to avoid the use of a large laparotomy wound. The advantages of this approach have been well documented, but so far the laparoscopic route has been little used by the general gynecologic surgical community. Although it has been possible in some units to reduce the laparotomy rate for hysterectomy to 10%, there is evidence from the United Kingdom and the United States that still more than 70% of all hysterectomies are still being performed by the laparotomy approach. This chapter seeks to explain why this should be so and how appropriate training and acquisition of the necessary skills in operative laparoscopy and vaginal surgery can be achieved to ensure that all patients who may benefit by avoiding a laparotomy incision are given the opportunity to do so. PMID- 10459061 TI - Laparoscopic colposuspension. AB - Laparoscopic Burch colposuspension has entered the surgical armory for the treatment of genuine stress incontinence. The limited data available of the outcomes of the procedure currently suggest that success rates are lower than for the open colposuspension in the short term. Data is lacking on long-term effectiveness. This and the longer operating times documented may reflect inexperience of surgeons. There seem to be advantages in terms of sooner return to normal voiding, earlier discharge from hospital, perioperative morbidity, and a lower incidence of voiding problems when compared with the open procedure. No reduction in the incidence of de novo detrusor instability postoperatively has emerged. A large randomized controlled trial of the two procedures is required to provide higher quality data on outcomes. PMID- 10459062 TI - Photodynamic cystoscopy for detection of bladder tumors. AB - The use of a photodynamic fluorescence marker for diagnosis of tumors is an intriguing concept to improve thoroughness of surgical tumor resection. Complete surgical resection of multifocal bladder tumors and flat lesions as carcinoma in situ is known to be difficult, and thus a source of recurrencies. We report on the recent experience with the intravesical application of the photosensitizer prodrug 5-aminolevulinic acid (5-ALA), which is a nontoxic physiological heme substrate. Initial results from fluorescence diagnosis using krypton laser light and recent results with a modified incoherent light source constantly showed a 25% increase in urothelial tumor detection compared with white light endoscopy. Because of the high sensitivity, the number of biopsies could be decreased constantly compared with routine random biopsies taken under white light endoscopy. The results show about 25% to 30% of cases with fluorescent lesions, which are histologically benign. 5-ALA is a promising tool for diagnosis of bladder cancer. The outcome of the initial study of 5-ALA in urology in Germany is positive and is continued by prospective multicenter clinical studies to prove the hypothesis of reduction of tumor recurrence with this method. 5-ALA-based fluorescence endoscopy is strongly recommended for further clinical testing. PMID- 10459063 TI - Laparoscopic management of benign ovarian disease. AB - There was little controversy in the management of ovarian cysts until 15 years ago when operative endoscopy gained popularity. Before this time, laparotomy was the standard form of treatment for pelvic masses. Simple cysts were treated laproscopically by aspiration and solid or large tumors with any malignant potential by laparotomy. Constantly improving technology and instrumentation has allowed more and more operations to be performed laparoscopically. The gynecological oncologist has been wary of adopting this form of surgery for potential gynecological malignancies because of the safety and efficacy issues. Pelvic pathology, particularly ovarian tumors, pose a difficult problem. Apart from difficulty in screening, it is also difficult to confirm the nature of an ovarian pathology using the currently available investigative modalities. If the nature of an ovarian cyst can be ascertained beyond doubt, then the treatment can be simple, laparoscopy for benign and laparotomy for malignant. Having said that, more enthusiastic operative laparoscopists are using this approach to treat gynecological malignancies. We discuss the role of laparoscopy in the management of ovarian cysts. PMID- 10459064 TI - Laparoscopic sterilization. AB - Laparoscopic female sterilization is still the leading method of family planning for patients who have completed their family. Mechanical methods include clips and rings and are preferred because they are safe and efficient and can be used on a day case basis. Appropriate training ensures improved results with fewer complications. Clips and rings have an improved reversal potential. PMID- 10459065 TI - Enteral nutrition in intensive care patients: a practical approach. AB - Severe protein-calorie malnutrition is a major problem in many intensive care (ICU) patients, due to the increased catabolic state often associated with acute severe illness and the frequent presence of prior chronic wasting conditions. Nutritional support is thus an important part of these patient's management. Over the years, enteral nutrition (EN) has gained considerable popularity, due to its favorable effects on the digestive tract and its lower cost and rate of complications compared to parenteral nutrition. However, clinicians caring for ICU patients are often faced with contradictory data and difficult decision making when having to determine the optimal timing and modalities of EN administration, estimation of patient requirements and choice of formulas. The purpose of this paper is to provide practical guidelines on these various aspects of enteral nutritional support, based on presently available evidence. PMID- 10459066 TI - Effects of dietary polyunsaturated fatty acids and nucleotides on tissue fatty acid profiles of rats with carbon tetrachloride-induced liver damage. AB - The deficiency of polyunsaturated fatty acids (PUFA) that occurs in plasma of patients with liver cirrhosis has been assessed in rats with severe steatosis and mild liver necrosis induced by repeated administration of low doses of carbon tetrachloride (CCl(4)). The contribution of both dietary (n-3) long-chain PUFA and nucleotides to the recovery of the altered fatty acid profiles of tissue lipids of these rats has also been studied. Two groups of rats were used. The first was intraperitoneally injected 0.15 ml of a 10% (v/v) CCl(4)solution in paraffin per 100 g of body weight, three times a week for 9 weeks; the second received paraffin alone. After the treatment, six rats of each group were killed. Afterwards, the remaining controls were fed a semipurified diet (SPD) for 3 weeks, and the remaining rats in the CCl(4)group were divided into three new groups: the first was fed the SP diet; the second was fed the SP diet supplemented with 1% (n-3) polyunsaturated fatty acids (PUFA diet); and the third was fed the SP diet supplemented with 250 mg nucleotides per 100 g diet (NT diet). Fatty acids of plasma, erythrocyte membranes and liver microsomes were analyzed. Decreases in linoleic and arachidonic acids in both total plasma lipids and liver microsomal phospholipids were the main findings due to CCl(4)treatment. The rats that received CCl(4)and the PUFA diet showed the lowest levels of (n-6) PUFA and the highest levels of (n-3) PUFA in liver microsomal phospholipids, as well as a significant increase of (n-3) PUFAs in total plasma lipids. The animals that received the NT diet showed no signs of fatty infiltration and exhibited the highest levels of (n-6) PUFAs in liver microsomal phospholipids. These results show that CCl(4)affects fatty acid metabolism which is accordingly reflected in altered tissue fatty acid profiles, and that balanced diets containing PUFA and nucleotides are important for the recovery of the damaged liver in rats. PMID- 10459067 TI - The thiamin, riboflavin and pyridoxine status of patients on emergency admission to hospital. AB - The aim of this study was to assess the prevalence of thiamin, riboflavin and pyridoxine deficiencies at admission to an acute hospital. One hundred and twenty adult patients were selected at random from those admitted via the Accident and Emergency department over 3 days. Comparisons were made with a group of 80 healthy blood donors sequentially attending a local transfusion centre. The alcohol intake of 500 patients admitted sequentially via the same Accident and Emergency department was also assessed. Erythrocyte transketolase (ETK), glutathione reductase (EGR) and aspartate aminotransferase (EAA) coenzyme activation assays were used to determine thiamin, riboflavin and pyridoxine deficiencies. The prevalences of deficiency states in the inpatient group were 21, 2.7 and 32% for thiamin, riboflavin and pyridoxine deficiencies respectively with 49.2% being deficient in one or more vitamin. The mean alcohol intake in the group of patients in whom this was assessed was 9.7 units per week compared with 10 units per week amongst blood donors. PMID- 10459068 TI - The effects of enteral tube feeding and parenteral nutrition on appetite sensations and food intake in health and disease. AB - Enteral tube feeding (ETF) and parenteral nutrition (PN) are unphysiological methods of feeding. They may not elicit the cephalic phase response because part or all of the gastrointestinal tract is bypassed, nutrients are typically given in liquid form by a continuous infusion over many hours and often overnight while patients sleep. Work conducted in animals, healthy subjects and patients suggests that nutrients delivered as ETF or PN are less effective in relieving appetite sensations than food intake. Distressing appetite sensations may even occur despite the provision by artificial nutrition of sufficient nutrients to meet requirements. The energy provided by ETF and PN is largely additional to oral food intake in humans eating ad libitum, although the extent to which this occurs may decrease with time. There is a need to establish ways (e.g. nutritional, pharmacological, psychological) to suppress appetite sensations and food intake when eating is contraindicated, and to enhance them when weaning from artificial nutrition is desirable. PMID- 10459069 TI - How often do parenteral nutrition prescriptions for the newborn need to be individualized? AB - BACKGROUND: Parenteral nutrition is commonly given in the newborn period to premature infants or those with gastrointestinal disorders. Computer-assisted prescribing is widely used, with prescriptions for each patient being varied on a daily basis. It has previously been suggested that 'individualization' of feeds may have little clinical benefit whilst increasing pharmacy workload and costs. However, the scope for use of standard feed solutions as an alternative remains uncertain. METHODS: To assess the potential for using standardized pre-mixed feeds we prospectively reviewed 148 computer assisted prescriptions for newborn infants in order to establish how often the prescribing clinician adhered to the computer protocol, and the reason for modification when this occurred. RESULTS: Only one-fifth of feeds were based strictly on the computer recommendation with no, or minimal, modification. However, many of the deviations in the other four fifths of feed prescriptions reflected a routine use of higher carbohydrate, sodium and phosphate intakes implying that a higher proportion of feeds could be 'standardized' if the computer regimens were modified to reflect current nutritional practices on the unit. CONCLUSIONS: This study suggests that the introduction of standard PN feeds could considerably reduce the use of computer assisted individualized PN prescriptions on the neonatal unit. The practical implications of such a system for pharmacy and the potential cost benefits deserve further investigation. PMID- 10459070 TI - Nutritional status at submission for dietetic services and screening for malnutrition at admission to hospital. PMID- 10459071 TI - Molecular regulation of protein catabolism in trauma patients. PMID- 10459072 TI - Effects of diet composition on triacylglycerol metabolism in humans. PMID- 10459073 TI - Glutamine, a regulator of acute phase protein synthesis. PMID- 10459074 TI - Peroxidation of lipid emulsions: effects of changes in fatty acid pattern and alpha-tocopherol content on the sensitivity to peroxidative damage. PMID- 10459075 TI - Preoperative oral carbohydrates and postoperative insulin resistance. AB - Infusions of carbohydrates before surgery have been shown to reduce postoperative insulin resistance. Presently, we investigated the effects of a carbohydrate drink, given shortly before surgery, on postoperative insulin sensitivity. METHODS: Insulin sensitivity and glucose turnover ([6, 6,(2)H(2)]-D-glucose) were measured using hyper-insulinemic, normoglycemic clamps before and after elective surgery. Sixteen patients undergoing total hip replacement were randomly assigned to preoperative oral carbohydrate administration (CHO-H, n = 8) or the same amount of a placebo drink (placebo, n = 8) before surgery. Insulin sensitivity was measured before and immediately after surgery. Patients undergoing elective colorectal surgery were studied before surgery and 24 h postoperatively (CHO-C (n = 7), and fasted (n = 7), groups). The fasted group underwent surgery after an overnight fast. In both studies, the CHO groups received 800 ml of an isoosmolar carbohydrate rich beverage the evening before the operation (100g carbohydrates), as well as another 400 ml (50g carbohydrates) 2 h before the initiation of anesthesia. RESULTS: Immediately after surgery, insulin sensitivity was reduced 37% in the placebo group (P < 0.05 vs. preoperatively) while no significant change was found in the CHO-H group (-16%, p = NS). During clamps performed 24h postoperatively, insulin sensitivity and whole-body glucose disposal was reduced in both groups, but the reduction was greater compared to that in the CHO-C group (-49 +/- 6% vs. -26 +/- 8%, P>> 0.05 fasted vs. CHO-C). CONCLUSIONS: Patients given a carbohydrate drink shortly before elective surgery displayed less reduced insulin sensitivity after surgery as compared to patients undergoing surgery after an overnight fast. PMID- 10459076 TI - HSP70 expression in granulocytes and lymphocytes of patients with polytrauma: comparison with plasma glutamine. AB - Heat shock proteins (HSP's) are a set of conserved proteins which confer tolerance to stress. These proteins play a major role in the pathophysiology of infection and inflammation. Induction of HSP's before onset of sepsis is able to reduce or prevent organ damage and death. GLN is known to influence the expression of HSP70 in different cell types. In this work we tried to find out if there is an association between plasma GLN levels and HSP70 expression in immune cells. We investigated six polytraumatized patients and a control group of six healthy donors. HSP70 expression was investigated by western blot analysis and immune-histochemistry. We demonstrated that granulocytes and lymphocytes behave differently in the expression of HSP70 in polytraumatized patients. In healthy donors both lymphocytes and granulocytes showed a pronounced expression of HSP70. In contrast, most of the polytraumatized patients showed no HSP70 expression in granulocytes. In lymphocytes of these patients, however, a pronounced expression similar to that of healthy volunteers was observed. Plasma glutamine levels were reduced in all patients and at normal range in healthy donors. These results suggest that lymphocytes and granulocytes behave different when confronted with a reduction of plasma GLN levels. PMID- 10459077 TI - Abnormalities in branched-chain amino acid metabolism in cirrhosis: influence of hormonal and nutritional factors and directions for future research. AB - Plasma branched-chain amino acid (BCAA) levels are decreased in patients with liver cirrhosis, owing to an increase in BCAA tissue uptake and/or catabolism and a decrease in BCAA production from proteins. Non-specific factors such as malnutrition worsen this picture. Studies of BCAA fluxes and protein turnover in cirrhotic patients have given conflicting results due to patient heterogeneity, differences in method and bias in the expression of results. In well compensated cirrhosis, muscle wasting is moderate and probably due more to decreased protein synthesis than to increased protein catabolism. Hyperinsulinemia has been suggested as the main cause of decreased BCAA levels, by increasing BCAA uptake in muscle and additionally in adipose tissue. However, as depletion of fat stores is frequent in cirrhosis, this effect is certainly quantitatively weak. Also, there is no correlation between state of hyperinsulinemia and decrease in BCAA levels. An effect of cytokines (IL1 and TNF) on muscle BCAA catabolism is a possibility. Until recently, the contribution of the liver to abnormal BCAA metabolism has been underestimated. In cirrhotic liver an increase in liver transamination of branched-chain keto acids (BCKAs) has been suggested and may result from inhibition of liver BCKA dehydrogenase. A modification of protein turnover in cirrhotic liver must be also considered. Lastly, the contribution of non-hepatocyte liver cells, which are activated in cirrhosis, remains to be assessed. PMID- 10459078 TI - Nutritional status at submission for dietetic services and screening for malnutrition at admission to hospital. AB - This paper presents two studies in a quality management project that aims to diminish malnutrition among hospitalized patients. The objective of study 1 was to investigate what information was available on the nutritional status of patients submitted for dietetic services for reasons other than obesity (n= 167) and of study 2 to evaluate a nine-question screening sheet for malnutrition in patients (n= 115) within 48 h of admission to the hospital. In study 1 sufficient data to evaluate nutritional status was found for 17% of the patients submitted for dietetic services. In study 2 the screening sheet identified 21% of the patients as malnourished and a full nutritional assessment of seven anthropometrical and biochemical measurements 20%. The screening sheet could be simplified to six questions and then had a sensitivity of 0.69, a specificity of 0.91 and a positive predictive value of 0.65. It is concluded that evaluation of nutritional status in hospitalized patients has been disregarded and a simple screening sheet can be used to identify patients in need of further nutritional assessment and treatment. PMID- 10459079 TI - Is alpha-ketoisocaproyl-glutamine a suitable glutamine precursor to sustain fibroblast growth? AB - BACKGROUND: Glutamine is considered an essential nutrient for cellular growth. AIM: To test the suitability of alpha-ketoisocaproyl-Gln (Kic-Gln) as a new glutamine (Gln) precursor to sustain human fibroblast growth. METHODS: [3H] thymidine uptake into cellular DNA of human fibroblasts. Extracellular and intracellular amino acid patterns were determined with peptides and acylated compounds. RESULTS: L-alanyl-L-glutamine (used here as a recognized Gln precursor) promoted DNA synthesis, while N-acetyl-L-glutamine (used here as a negative control since it is known to be a poor Gln precursor) and alpha ketoisocaproyl-glutamine had no effect. Alanyl-glutamine progressively gave rise to free glutamine in the growth medium. In contrast, glutamine supplied in acylated form was poorly available and did not appear in free form in the medium. In addition, only alanyl-glutamine increased intracellular glutamine and glutamate levels. In contrast, Kic-Gln was able to sustain net protein synthesis as judged by total protein content and reduced intracellular levels of most essential amino acids. CONCLUSION: Kic-Gln appears to be a poor extra-cellular precursor of Gln to sustain cell growth. PMID- 10459080 TI - Bioelectrical impedance analysis to assess changes in total body water in patients with cancer. AB - Predominantly based on studies in obese individuals, the applicability of single frequency bioelectrical impedance analysis (BIA) to measure changes in total body water and fat-free mass has been questioned. To further clarify this issue, we compared changes in BIA-derived height(2)/resistance (ht(2)/R) with changes in total body water (deuterium dilution, delta-TBWdeu) in cancer patients participating in a clinical trial. Thirty-three patients (mean body mass index 23.2 +/- 3.5 kg/m(2)) were studied after an average follow-up of 11 weeks. Changes in TBWdeu occurred in both directions (mean +0.2 +/- 1.6 L, range -3.3 to +3.1 L). These changes were significantly predicted by changes in ht(2)/R (r(2)0.43, P < 0.0001, SEE 1.22 L), although precision was poor (residual SD 1. 2 L). There were in this regard no significant differences between patients with and without underweight. We conclude that in underweight and normal-weight cancer patients, BIA-derived changes in ht(2)/R significantly predict changes in total body water assessed by deuterium dilution. PMID- 10459081 TI - Knowledge about the assessment and management of undernutrition: a pilot questionnaire in a UK teaching hospital. AB - BACKGROUND & AIMS: The detection, prevention and treatment of undernutrition in hospitals is often poor. This study assesses the knowledge about undernutrition of staff in a UK teaching hospital. METHODS: Twenty nine doctors, 65 final year medical students, 45 nurses, 11 dietitians, and 11 pharmacists anonymously completed a questionnaire of 20 multiple choice questions. One of five possible answers was considered correct. Twelve questions were about adult nutritional assessment and requirements, five about oral/enteral nutrition and three about parenteral nutrition. RESULTS: Dietitians scored significantly more (median 16) than the other groups (doctors: seven, medical students: eight, nurses: seven and pharmacists: nine) (P < 0.0001). Medical students scored more than doctors (P < 0.001). Examples of areas in which knowledge could be improved are: 67% respondents thought the prevalence of hospital undernutrition to be less than 30%. While 91% of respondents correctly chose a well 70 kg man to need about 2000 kcal/day, only 23% knew that approximately the same amount was needed for a febrile post-operative patient. Sixteen percent knew antibiotic treatment to be the most common reason for enteral feeding-related diarrhoea. CONCLUSIONS: Knowledge about the assessment and management of undernutrition among doctors, medical students, nurses and pharmacists was poor. This questionnaire provides a framework for teaching and auditing the effectiveness of an educational program. PMID- 10459082 TI - Incidence of vein thrombosis in peripheral intravenous nutrition: effect of fat emulsions. AB - The relative risk of thrombophlebitis induced by either Long-chain Triglycerides (LCTs) or Medium-chain Triglycerides (MCTs) during peripheral i.v. nutrition (PIN) was evaluated. A total of 76 patients were randomly assigned into group A (n=40) and group B (n=36). The nutritional requirements in both groups were covered by a standardized regime of osmolality 1130 mOsm and pH 5.2, which provided 14 kg/day(-1) nitrogen, 600 kcal/day(-1) of carbohydrates and 1000 kcal/day(-1) of lipids. Group A received the lipids as pure LCTs while group B received a mixture of LCTs/MCTs at a ratio 1:1. The infused nutritional volume was 2000 ml and was delivered via a suitable vein in a proximal forearm, using a fine bore polyurethane 22G catheter. The two standardized regimes were evaluated over a 10 day period regarding the incidence of thrombophlebitis. The cumulative risk of thrombophlebitis was documented to be significantly lower in group A compared to group B (17.5% versus 44.4%, P < 0.05). LCTs appear to prolong peripheral vein feeding by lessening the reaction of venous endothelium to the irritating nutritional infusate. PMID- 10459083 TI - The Swedish version of the patient-generated subjective global assessment of nutritional status: gastrointestinal vs urological cancers. AB - PURPOSE: To translate and evaluate the Patient-Generated Subjective Global Assessment (PG-SGA) a method for the assessment of nutritional status. METHOD: Eighty-seven patients with gastrointestinal and urological tumours completed four sections and the remaining sections were independently completed by a doctor and a dietician. Patients were classified as SGA A (well nourished), SGA B (moderately/suspected of being malnourished) or SGA C (severely malnourished). RESULTS: Interobserver agreement was complete in 90% of the cases. More patients with gastrointestinal cancers than with urological cancers were classified as SGA B and C. Mean levels of S-albumin and P-prealbumin differed between the SGA classes. Multivariate logistic regression analyses showed independent contributions to the overall classification by weight loss in the last 6 months, level of food intake, problems with eating, physical activity and muscle wastage. Survival was significantly higher in SGA A than in SGA B+C, P < 0.001. CONCLUSION: The PG-SGA is useful for the assessment of nutritional status. Patients had no problems in answering the questions. The PG-SGA also carried prognostic information. PMID- 10459084 TI - Early response of ornithine decarboxylase activity and energy metabolism to postsurgery refeeding in rat small intestine. AB - INTRODUCTION: Enterocyte proliferation and cellular energy status are important to intestinal integrity after starvation and trauma. The proliferative response to nutrients is expressed in the activity of ornithine decarboxylase (ODC), but ODC activity and ATP level in the intestinal mucosa the first hours after surgery and immediate refeeding are not known. METHODS: Male Wistar rats (240-280 g) were starved for 48 h and submitted to laparotomy with distal ileal transection, gastrostomy and jejunal instillation of either enteral formula or saline. The ODC activity and ATP content of the jejunal mucosa were analysed in samples taken at 1, 2, 4 and 6 h after surgery. RESULTS: ODC activity increased and reached the highest peak at 2 h in the refed animals. ATP concentration and energy charge of jejunal mucosa were significantly reduced 6 h after surgery compared to initial levels, but there were no differences between animals that were refed or not. Intestinal transection did not stimulate ODC activity. CONCLUSION: ATP levels in intestinal mucosa decreased after surgery, and early enteral feeding did not seem to prevent this decrease during the first 6 h. Refeeding immediately after surgery elicits an early but transient increase of ODC activity in rat jejunal mucosa. PMID- 10459085 TI - Editorial PMID- 10459086 TI - Predicting the relative efficacy of shock waveforms for transthoracic defibrillation in dogs. AB - STUDY OBJECTIVE: Previous work has shown that a passive membrane model using a parallel resistor-capacitor circuit is capable of predicting optimal waveforms for transvenous defibrillation. This study tested the ability of that model to predict optimal waveforms for transthoracic defibrillation. METHODS: This study was divided into 3 parts, each of which determined transthoracic defibrillation thresholds (DFTs) in 6 dogs for several different waveform shapes and durations. For each part, strength-duration relationships were determined from both experimental and model data and then compared with test model predictions. Part 1 DFTs were determined at various durations for 3 different monophasic waveforms the ascending ramp, descending ramp, and square waveform. Part 2 DFTs were determined for 3 biphasic waveforms. Phase 1 was a 30-ms ascending ramp, and phase 2 was an ascending ramp, a descending ramp, or a square waveform. Part 3 DFTs were determined for 3 biphasic waveforms with very short second-phase durations. Phase 1 was a 30-ms ascending ramp, and phase 2 was a descending ramp. RESULTS: For part 1, the model was able to predict the relative defibrillation efficacy of the 3 monophasic waveforms ( P < .05). For parts 2 and 3, the model was able to predict the biphasic waveforms with the lowest DFTs. These predictions were based on the criterion that the model response at the end of the second phase should return to or slightly pass the model response value at the beginning of the first phase. CONCLUSION: The resistor-capacitor model successfully predicted the relative defibrillation efficacy of several different waveforms delivered transthoracically. PMID- 10459087 TI - Effect of an emergency department asthma program on acute asthma care. AB - STUDY OBJECTIVE: To examine the effect of an emergency department program on acute asthma care. METHODS: We conducted a before-after study of an acute asthma quality improvement initiative in an urban teaching hospital with 65,000 annual ED visits. In mid-1994, a multidisciplinary group identified deficiencies in acute asthma care, developed and implemented a local version of the National Asthma Education Program's practice guidelines (including a standard asthma order sheet), and provided new peak flow (PF) meters. The "before" group comprised all adults with acute asthma seen during January 1994 (n=51); "after" groups comprised all adults with acute asthma seen during October 1994, February 1995, and June 1995 (n=145). Data were compared across months using a nonparametric test for trend. RESULTS: Although patient demographic characteristics and asthma severity were similar across months, ED process of care significantly changed. Initial PF measurements were obtained in 20% of patients before intervention, compared with 82%, 84%, and 83% during the postintervention months ( P for trend <.001). Follow-up PF readings were obtained in 22%, 70%, 78%, and 62% ( P <.001). Median delays to beta-agonist and steroid therapy decreased by approximately 16 minutes ( P <.001) and 34 minutes ( P =.04), respectively. Outcomes improved, with median ED length of stay decreasing by 58 minutes ( P =.01), and fewer inpatient admissions ( P =.05); there was no significant change in 4-week relapse to our hospital. CONCLUSION: A guideline-based ED asthma program changed clinical practice and improved acute asthma care in a sustained fashion. The effect of this intervention on cost and other outcomes is uncertain. PMID- 10459088 TI - A 5-year time study analysis of emergency department patient care efficiency. AB - STUDY OBJECTIVES: We conducted a 5-year time study analysis of emergency department patient care efficiency. Our specific aims were (1) to calculate the main ED patient care time intervals to identify areas of inefficiency, (2) to measure the effect of ED and inpatient bed availability on patient flow, (3) to quantitatively assess the effects of administrative interventions aimed at improving efficiency, and (4) to evaluate the relationship between waiting times to see a physician and the number of patients who leave without being seen (LWBS) by a physician. METHODS: Seven 1-week ED patient flow time studies were conducted from September 1993 to July 1998 using identical study design and methodology. Patients presenting with complaints of chest pain, abdominal pain, vaginal bleeding, and extremity injury were included to represent the level of severity of patient conditions seen in our Los Angeles County hospital ED. The calculated time intervals representing the main phases of evaluation and treatment were (1) triage presentation to completion of registration, (2) completion of registration to ED treatment area entry, (3) ED treatment area entry to initial medical assessment, (4) triage presentation to initial medical assessment, (5) initial medical assessment to disposition order, and (6) disposition order to patient discharge from the ED. Total ED lengths of stay (LOS) were also calculated as overall measures of efficiency. Time intervals were compared depending on the availability of ED and hospital inpatient beds. The effects of administrative interventions on the specific time intervals were assessed. The relationship between the median waiting time to see a physician and the number of LWBS patients was evaluated. Administrative interventions were implemented by a special interdepartmental continuous quality improvement committee. Interventions were aimed at specific sources of delay and inefficiency identified by the time studies. RESULTS: Eight hundred twenty-six patients were included in the 7 time studies. The unavailability of ED and inpatient beds was associated with significant delays. There was a significant reduction of the median total ED LOS from 6.8 hours to 4.6 hours over the first 5 periods, presumably resulting from the administrative interventions. Median total ED LOS, however, increased from 4.6 hours to 6.0 hours during the last 2 periods, possibly as a result of an increase in our ED patient census and reductions in both nursing and physician staffing imposed by the recent Los Angeles County fiscal crisis. The number of LWBS patients was closely correlated to waiting time to see a physician ( r =0.79, beta=5.20, P =.033). CONCLUSION: Time studies are an effective method of identifying areas of patient care delay. In our ED, targeted administrative interventions apparently reduced the total ED LOS and improved overall efficiency. Despite initial decreases in ED LOS, efficiency appeared to be adversely affected by reductions in nursing and physician staffing and increases in our patient census. The strength of the relationship between waiting times to see a physician and the number of LWBS patients suggests that decreasing waiting times may reduce the number of LWBS patients. PMID- 10459089 TI - Mandatory reporting laws do not deter patients from seeking medical care. AB - STUDY OBJECTIVE: As of March 1994, 45 states had laws that, to varying extents, required health practitioners to report cases of domestic violence (DV). Colorado passed a mandatory DV reporting law in 1995. Laws that mandate police involvement in cases of DV injuries have been criticized because of concerns that these laws deter victims from seeking medical care. We hypothesized that these laws would deter DV victims from seeking medical care. METHODS: A questionnaire was administered in 3 stages: stage 1, convenience time blocks at 2 emergency departments and a primary care clinic; stage 2, prospective randomized blocks at an inner-city ED; and stage 3, a targeted population of women at risk for DV. All English-speaking, noncritical adult patients who presented during the time blocks were eligible to participate. RESULTS: Five hundred seventy-seven patients participated; 55% of the patients were aware of the mandatory DV reporting law. Twenty-seven percent of the patients would be more likely to seek medical care because of this law. Only 12% of patients stated that they would be less likely to seek medical care for a DV-related injury because of this law (15% of men and 9% of women; P =.001). There was no difference between ED patients and targeted female patients at risk for DV in seeking medical care ( P =.833). CONCLUSION: Only rarely did mandatory reporting laws appear to adversely affect a patient's decisions to seek medical care in this study. The benefits of mandatory reporting must be measured to assure that they justify deterrence to a small minority of patients. PMID- 10459090 TI - Screening for ketonemia in patients with diabetes. AB - STUDY OBJECTIVES: To determine the sensitivity and specificity of the urine ketone dip test as a screening test for ketonemia in hyperglycemic patients and to compare the performance of the urine ketone dip test with the anion gap and serum bicarbonate level. METHODS: This was a prospective study conducted in an urban, university-affiliated public hospital emergency department. Inclusion criteria consisted of (1) patients with known diabetes and hyperglycemia (glucose level>200 mg/dL) and any complaint of illness, or (2) patients with hyperglycemia and symptoms of undiagnosed diabetes mellitus. Urine ketone dip test, serum ketone, and electrolyte levels were determined on all subjects. Sensitivity, specificity, and predictive values along with 95% confidence intervals (CIs) were calculated. RESULTS: The study group comprised 697 patients, including 98 patients with diabetic ketoacidosis (DKA) and 88 with diabetic ketosis (DK). The sensitivity, specificity, positive, and negative predictive values of the urine ketone dip test for the detection of DKA were 99% (95% CI 97% to 100%), 69% (95% CI 66% to 73%), 35% (95% CI 29% to 41%), and 100% (95% CI 99% to 100%), respectively. For DKA and DK, the sensitivity, specificity, positive, and negative predictive values of the urine ketone dip test were 95% (95% CI 90% to 97%), 80% (95% CI 76% to 83%), 63% (95% CI 57% to 69%) and 98% (95% CI 96% to 99%). The anion gap and serum bicarbonate level were less sensitive but more specific than the urine ketone dip test for the detection of DKA and DK. CONCLUSION: The urine ketone dip test has high sensitivity for detecting DKA and a high negative predictive value for excluding DKA in hyperglycemic patients with diabetes with any symptoms of illness. The urine ketone dip test is a better screening test for DKA and DK than the anion gap or serum bicarbonate. PMID- 10459091 TI - Warmed versus room temperature saline solution for ear irrigation: a randomized clinical trial. AB - STUDY OBJECTIVE: To test the hypothesis that ear irrigation with warmed normal saline solution is more comfortable and results in fewer side effects than irrigation with room temperature saline solution in normal volunteers. METHODS: The study was a randomized, single-blind, crossover trial in which each subject received 30 mL warmed normal saline solution in 1 ear and 30 mL room temperature saline solution in the opposite ear. The solutions (warmed versus room temperature) and the order of irrigation (right versus left ear) were separately randomized. Investigators obtaining scores were blinded to solution temperature. Subjects rated the discomfort of irrigation, using separate visual analog scales, from 0 (no pain) to 100 mm (worst pain ever). RESULTS: Forty volunteers were enrolled in the study. The mean difference in visual analog scale scores favoring warmed over room temperature saline solution was 26 mm (95% confidence interval [CI], 19 to 33 mm; P <.0001). Twenty percent more patients reported dizziness with room temperature irrigation (95% CI, 6% to 34%). There was no gender effect or order effect for the 2 solutions. CONCLUSION: Warmed normal saline solution was both clinically and statistically more comfortable than room temperature saline solution as an ear irrigant in normal volunteers. Significantly less dizziness was reported with the warmed solution. PMID- 10459092 TI - Common presentations of amebic liver abscess. AB - STUDY OBJECTIVE: The most common extraintestinal manifestation of Entamoeba histolytica, the agent of amebiasis, is a hepatic abscess. This infection is common throughout the world and can be associated with life-threatening consequences. Given the often nonspecific nature of the complaints related to an amebic abscess, a retrospective review of patients with confirmed disease was done to recognize the most common patterns of presentation. METHODS: A retrospective case series was conducted of all patients with confirmed amebic liver abscess over a 5-year period. All available emergency department and inpatient records were reviewed. Age, sex, country of origin, chief complaint (including duration), vital signs, and physical and laboratory findings were recorded. The use of ultrasonography, computed tomography scan, chest radiograph, and serum antibodies was noted, as well as the final ED diagnosis. RESULTS: Seventy-five patients were reviewed; mean patient age was 35.5 years, 80% were male, and Mexico was the country of origin for 64%. The most common complaint was fever (77%), followed by abdominal pain (72%), which was most often located in the right upper quadrant. Cough (16%), chest pain (19%), and chest radiographic abnormalities (57%) were also common. The majority of patients (69%) had symptoms for less than 13 days. The WBC count was the most consistent laboratory abnormality (83%), whereas the liver aminotransferase, alkaline phosphatase, and bilirubin levels were often normal. Most patients received their diagnoses on the basis of ultrasonography (85%), followed by a confirmatory serum antibody titer (88%). The diagnosis of amebic liver abscess was correctly made in the ED in 31.5% of the patients, with the most common misdiagnoses being cholecystitis (16.4%), hepatitis (12.3%), and pneumonia (9.6%). CONCLUSION: Patients with amebic liver abscess do present to EDs in the southwestern United States, especially in areas with a high immigrant population from endemic areas. Patients with complaints of fever and right upper quadrant abdominal pain, especially men of Hispanic origin, warrant a high degree of vigilance. Whereas most laboratory studies are unhelpful, the diagnosis can often be made in the ED by means of a bedside ultrasonographic test. Treatment should be initiated with metronidazole with disposition to an inpatient medical service. PMID- 10459093 TI - Laceration management. AB - In 1996, almost 11 million lacerations were treated in emergency departments throughout the United States. Although most lacerations heal without sequelae regardless of management, mismanagement may result in wound infections, prolonged convalescence, unsightly and dysfunctional scars, and, rarely, mortality. The goals of wound management are simple: avoid infection and achieve a functional and aesthetically pleasing scar. Recent US Food and Drug Administration approval of tissue adhesives has significantly expanded clinicians' wound closure options and improved patient care. We review the general principles of wound care and expand on the use of tissue adhesives for laceration repair. PMID- 10459094 TI - Baby steps. PMID- 10459095 TI - Human error in emergency medicine. PMID- 10459096 TI - The potential for improved teamwork to reduce medical errors in the emergency department. The MedTeams Research Consortium. AB - This article describes emergency department care work teams designed to improve team communication and coordination and reduce error. The core of this teamwork system is the teaching of teamwork behaviors and skills, development of teamwork habits, and creation of small work teams, all of which are key teamwork concepts largely drawn from successful aviation programs. Arguments for enculturating teamwork into ED practice are drawn from a retrospective study of ED malpractice incidents. Fifty-four incidents (1985-1996), a sample of convenience drawn from 8 hospitals, were identified and judged mitigable or preventable by better teamwork. An average of 8.8 teamwork failures occurred per case. More than half of the deaths and permanent disabilities that occurred were judged avoidable. Better teamwork could save nearly $3.50 per ED patient visit. Caregivers must improve teamwork skills to reduce errors, improve care quality, and reduce litigation risks. PMID- 10459097 TI - Motor vehicle safety: current concepts and challenges for emergency physicians. AB - Motor vehicle travel is the primary means of transportation in the United States, providing freedom in travel and enterprise for many people. Motor vehicle crashes are the leading cause of death for the young and contribute to a high degree of morbidity and mortality for all ages. Motor vehicle crashes produce an enormous burden for society in terms of suffering, disability, death, and costs. Motor vehicle crash injury prevention is developing as a focused discipline to implement proven interventions involving technology and behavior known to prevent or reduce the severity of motor vehicle crash injury. Emergency physicians have an important role in advocating motor vehicle safety and injury prevention, both in the emergency department and within the community. PMID- 10459098 TI - Hypoketotic hypoglycemic coma in a 21-month-old child. AB - We present the case of a 21-month-old child with hypoketotic hypoglycemic coma. The differential diagnosis initially included metabolic causes versus a toxicologic emergency (unripe ackee fruit poisoning). Using information obtained from the emergency department, the diagnosis was confirmed as the late-onset form of glutaric acidemia type II. This case illustrates the importance of emergency physicians in the diagnosis and management of children with inborn errors of metabolism. PMID- 10459099 TI - Hypocalcemic tetany in a toddler with undiagnosed rickets. AB - A 17-month-old black girl presented to the pediatric emergency department with sudden onset of her hands and feet "drawing up." A diagnosis of vitamin D deficiency rickets with tetany was made on a history of exclusive breast-feeding, low serum calcium, phosphorus, and 25-hydroxyvitamin D levels, and bone radiographs. Nutritional rickets should be considered in the differential diagnosis of acute hypocalcemia. PMID- 10459100 TI - National Highway Traffic Safety Administration (NHTSA) notes. Child safety seat distribution: what works? PMID- 10459101 TI - Child safety seats--improving use step-by-step. PMID- 10459102 TI - Fifty-seven-year-old female. PMID- 10459103 TI - Glycoprotein IIb/IIIa inhibitors. PMID- 10459104 TI - Glycoprotein IIb/IIIa Inhibitors. PMID- 10459105 TI - Reporting of work-related injury or illness to OSHA. PMID- 10459107 TI - Pathogenesis of Envenomation. PMID- 10459106 TI - High temperature with cerebral hemorrhage. PMID- 10459108 TI - Appropriate utilization of air medical transport in the out-of-hospital setting. EMS Committee. PMID- 10459109 TI - Emergency physician contractual relationships. Emergency Medicine Practice Committee. PMID- 10459110 TI - Support for National Disaster Medical system. EMS Committee. PMID- 10459111 TI - Cutaneous laser resurfacing. AB - Cutaneous resurfacing with the new generation of carbon dioxide and erbium lasers has recently come into favor for the treatment of facial rhytides, photodamage, and scarring. The precise control of these resurfacing lasers over the extent of tissue vaporization minimizes thermal damage to the skin while maximizing therapeutic efficacy. Proper use of resurfacing lasers is contingent upon a complete understanding of their clinical, histologic, and ultrastructural effects, as well as an appreciation of the principles of laser safety. An organized approach to the preoperative, intraoperative, and postoperative management of the patient undergoing laser resurfacing will be provided, including a discussion of prevention and treatment of postoperative side effects and complications. (J Am Acad Dermatol 1999;41:365-89.) LEARNING OBJECTIVE: At the conclusion of this learning activity, participants should be familiar with the clinical, histologic, and ultrastructural effects of resurfacing lasers and be able to discuss the preoperative, intraoperative, and postoperative management of patients undergoing laser resurfacing. PMID- 10459112 TI - Paraneoplastic pemphigus associated with Castleman tumor, myasthenia gravis and bronchiolitis obliterans. AB - BACKGROUND: Cases of paraneoplastic pemphigus (PNP) have been reported associated with various lymphoproliferative malignancies and benign Castleman tumors, with the most severe course and fatal outcome seen in patients with bronchiolitis obliterans. OBJECTIVE: The aim was to establish immunologic associations by coexistence of Castleman tumor, myasthenia gravis, and bronchiolitis obliterans and to evaluate the treatment modalities. METHODS: Clinical studies included computed tomography of the mediastinum, computed tomography and magnetic resonance imaging of the abdominal cavity, and quantitative electromyography. Direct and indirect immunofluorescence on various substrates, immunoblot analysis, immunoprecipitation, and specific enzyme-linked immunosorbent assay using recombinant desmogleins (Dsg) were performed as immunologic assays. RESULTS: Direct and indirect immunofluorescence including rat bladder showed intercellular antibodies. Immunoblotting disclosed antibodies to envoplakin (210 kd protein) and periplakin (190 kd protein); in addition, immunoprecipitation detected antibodies to desmoplakin I (250 kd protein). Antibodies to Dsg3 (pemphigus vulgaris antigen) were detected by specific enzyme-linked immunosorbent assay. Myasthenia gravis was controlled by drugs; however, mucocutaneous changes were not fully responsive to corticosteroids and cyclophosphamide pulses, cyclosporine, and intravenous immunoglobulins. The surgical removal of Castleman tumor did not change the course of the disease. The fatal outcome was the result of bronchiolitis obliterans that occurred after the surgery and was only transitionally controlled by plasmapheresis. CONCLUSION: This is the first case of paraneoplastic pemphigus associated with Castleman tumor, myasthenia gravis, and bronchiolitis obliterans. Despite a benign character of the tumor the patient died, as do all patients with bronchiolitis obliterans. Massive plasmapheresis has only a transient effect. We confirmed the presence of antibodies to Dsg 3, in addition to the set of specific paraneoplastic pemphigus antibodies against various proteins of plakin family. PMID- 10459113 TI - Psoriasis causes as much disability as other major medical diseases. AB - BACKGROUND: Little is known about how the health-related quality of life (HRQL) associated with psoriasis compares with that of other patient populations. OBJECTIVE: We describe HRQL associated with psoriasis and compare it with HRQL of patients with other major chronic health conditions. A second aim is to identify which specific aspects of psoriasis contribute most to HRQL. METHODS: A total of 317 patients completed a non-disease-specific measure of HRQL. Responses were compared with those of patients with 10 other chronic health conditions. HRQL was regressed on ratings of 18 aspects of psoriasis. RESULTS: Patients with psoriasis reported reduction in physical functioning and mental functioning comparable to that seen in cancer, arthritis, hypertension, heart disease, diabetes, and depression. Six aspects of psoriasis predicted physical functioning, and 5 different disease aspects predicted mental functioning. CONCLUSION: The impact of psoriasis on HRQL is similar to that of other major medical diseases. Different aspects of psoriasis are related to the different dimensions of HRQL supporting the need for multidimensional treatment models. PMID- 10459114 TI - Half-side comparison of erythemogenic versus suberythemogenic UVA doses in oral photochemotherapy of psoriasis. AB - BACKGROUND: Early American and European multicenter trials on the efficacy of photochemotherapy (PUVA) for psoriasis have clearly shown that the treatment protocol has a crucial impact on the cumulative UVA dose required for clearing patients. Most, if not all, treatment protocols rely on the PUVA-induced erythema as a guideline for UVA dosimetry. OBJECTIVE: Our aim was to investigate whether phototoxic erythema is integral to an optimized PUVA protocol or reflects an unnecessary overexposure of patients. METHODS: A standard high-dose UVA regimen using minimal phototoxic doses (MPD) was compared against two different low-dose regimens. To this purpose a bilateral comparison study was performed on 31 patients and divided in two parts. In the first trial on 14 patients, half of each patient's body was irradiated at each visit with 1 MPD, whereas the other half received only two thirds of the MPD. In the second trial on 17 patients treatment with 1 MPD was compared against treatment with one half of the MPD. RESULTS: A total of 27 patients (12 patients in the first trial, 15 patients in the second trial) completed the study. In both trials the suberythemogenic doses were therapeutically as effective as the minimal phototoxic doses. CONCLUSION: We conclude that PUVA-induced erythema is not a prerequisite for effective psoriasis treatment and that a low-dose UVA regimen is a promising approach to increase the short- and long-term safety of photochemotherapy. PMID- 10459115 TI - A randomized paired comparison of photodynamic therapy and topical 5-fluorouracil in the treatment of actinic keratoses. AB - BACKGROUND: Photodynamic therapy (PDT) has not been compared with topical 5 fluorouracil (5-FU) in the treatment of epidermal dysplasia. OBJECTIVE: The purpose of this study was to assess the efficacy and tolerability of these two treatment modalities in 17 patients with actinic keratoses on the backs of the hands. METHODS: Each patient's right and left hands were randomized to receive either a 3-week course of topical 5-FU applied twice per day or PDT using topical 5-aminolevulinic acid (5-ALA) and then, after 4 hours, irradiation with an incoherent light source consisting of a 1200 W metal halogen lamp emitting red light (580 to 740 nm). Each hand randomized for PDT received 150 J/cm(2). The observed median fluence rate was 86 mW/cm(2) (interquartile range, 53 to 100 mW/cm(2)). All patients were reviewed at 1, 4, and 24 weeks after starting treatment. RESULTS: Fourteen of 17 patients (82%) completed the study. The mean lesional area treated with topical 5-FU decreased from 1390 mm(2) (standard deviation [SD], 1130) to 297 mm(2) (SD, 209). This represents a mean reduction in lesional area of 70% (confidence interval [CI], 61%-80%). The mean lesional area treated with topical PDT decreased from 1322 mm(2) (SD, 1280) to 291 mm(2) (SD, 274), representing a mean reduction in lesional area of 73% (CI, 61%-84%). The reduction in lesional area elicited by the two treatment methods was similar (CI, -25% to 17%). There was no statistically significant difference between the treatment methods in overall symptom scores for pain and redness. CONCLUSION: One treatment with PDT using topical 5-ALA appears to be as effective and well tolerated as 3 weeks of twice-daily topical 5-FU, a cheap and widely available alternative. PMID- 10459116 TI - Lupus erythematosus-associated red lunula. AB - BACKGROUND: Red lunulae have only rarely been described in patients with lupus erythematosus. OBJECTIVE: We assessed the frequency and classified the type of red lunulae in patients with definite lupus erythematosus seen in an interdisciplinary dermatorheumatologic outpatient clinic. METHODS: We studied 56 patients with either systemic or cutaneous lupus erythematosus for the presence of red lunulae. RESULTS: Eleven of 56 patients (19.6%) with lupus erythematosus had red lunulae. All of them showed a complete type of red lunulae, which was seen on all finger nails in 10 patients and on a single finger nail in 1 patient. Seven patients suffered from systemic lupus, the other from subacute cutaneous (n = 2) or chronic discoid cutaneous lupus (n = 2). There was no statistically significant difference in autoantibody expression or treatment regimen between patients with or without red lunulae. However, symptomatic patients had a shorter disease interval, and all but 1 showed either periungual erythema or chilblain lupus. Periungual erythema was not observed in any patient without red lunulae. CONCLUSION: Red lunulae, although rarely described in the literature, are not an uncommon symptom of patients with lupus erythematosus. They seem to be associated with periungual erythema or chilblain lupus. Red lunulae should be considered in the clinical spectrum of lupus disease. PMID- 10459117 TI - Oral lesions and symptoms related to metals used in dental restorations: a clinical, allergological, and histologic study. AB - BACKGROUND: Allergy to mercury as a cause of oral lichenoid lesions (OLL) remains controversial. Some authors reported high frequency of sensitization to mercury and beneficial effect from removal of amalgam fillings in such patients, whereas others state that this procedure affects favorably all OLL, whether patients are sensitized to inorganic mercury or not. OBJECTIVE: Our purpose was to determine the frequency of sensitization to metal salts in 194 patients (patients with OLL partly adjacent to amalgam fillings: 19, oral lichen planus (OLP) without close contact to amalgam: 42, other oral diseases: 28, oral complaints: 46, control group: 59). We further studied the histologic changes of biopsy specimens from positive patch tests to metal salts, and investigated the effect of removal of amalgam in OLL, to clarify whether it is possible to identify patients who will benefit from this procedure. METHODS: Patch testing was performed with the German standard series, a dental prosthesis series, and a metal salt series including gold, mercury, and palladium salts as well as other salts of metals used in dental restorations. Late readings (10 and 17 days after application of the patch tests) were performed in all patients. RESULTS: Of 19 patients with OLL adjacent to amalgam fillings, 15 (78.9%) were sensitized to inorganic mercury (INM), significantly more than those with OLL not adjacent to amalgam, other oral diseases or complaints, and the control group. In 5 of 15 (33.3%) of the patients with OLL, a positive patch test to INM was observed only at D10 or D17. Amalgam was removed in 18 patients with OLL (sensitization to INM: 15), and in 11 patients with OLP (sensitization to INM: 2). After removal, the lesions of 13 of 15 of the INM-sensitized patients with OLL (86. 7%) and 2 with OLP healed or improved significantly, but this was not observed with the INM negative patients. Frequency of sensitization to gold sodium thiosulfate (GST) and palladium chloride 1% pet (PDC) was high in all groups. This was partly because readings were performed late. Lesions of 2 patients with allergic contact stomatitis caused by gold and 1 caused by palladium healed completely after removal of these restorations. Histologically, lichenoid changes were observed in 14 of 36 biopsy specimens of positive patch tests from INM (9/21), GST (2/10), and PDC (3/5) in all patient groups, mainly in persistent patch tests at D10 or D17. This was not observed in 12 biopsy specimens taken from persistent patch tests from other substances, including nickel sulfate. CONCLUSION: Our results suggest that sensitization to mercury is an important cause of OLL, whether all lesions or only a part of them are adjacent to amalgam fillings. Sensitization to GST may reflect true gold allergy and should be considered as a cause of oral diseases in some patients. Sensitization to PDC is frequent but has yet only little clinical relevance. Patch tests may be positive only at D10 or D17. This suggests the importance of additional readings of GST, PDC, and mercury salts at this time. PMID- 10459118 TI - Oral nalmefene therapy reduces scratching activity due to the pruritus of cholestasis: a controlled study. AB - BACKGROUND: Intravenous naloxone frequently ameliorates the pruritus of cholestasis, but its low oral bioavailability precludes its use as a long-term therapy. Nalmefene is an orally bioavailable opiate antagonist. OBJECTIVE: We assessed the efficacy of oral nalmefene in ameliorating the pruritus of cholestasis. METHODS: In a prospective controlled study conducted in a tertiary referral hospital, 11 patients with generalized pruritus complicating chronic liver disease were randomized to receive either nalmefene or placebo in a double blinded fashion for 2-month periods. Scratching activity was measured continuously for 24-hour periods at baseline and at the end of each treatment period. RESULTS: Data on 8 patients who received at least 1 course of nalmefene were available for comparison with corresponding control data, which consisted of observations obtained during a course of placebo and/or at baseline. Nalmefene therapy was associated with a 75% reduction in the geometric mean hourly scratching activity (P <.01) and a decrease in the mean of a visual analogue score of the perception of pruritus in all 8 patients (mean decrease 77%, P <.01). CONCLUSION: Oral administration of nalmefene can ameliorate pruritus complicating chronic liver disease. PMID- 10459119 TI - Eyelid dermatitis to red face syndrome to cure: clinical experience in 100 cases. AB - A retrospective review of all eyelid dermatitis patients seen over an 18-year period revealed a large subgroup of patients who had, as the basis for their ongoing problem, an addiction to the use of topical or systemic corticosteroids. This group of 100 patients often sought many consultations with various physicians. Unrelenting eyelid or facial dermatitis often resulted in the use of increasing amounts of corticosteroids for longer periods of time. Soon the skin became addicted. Once the work-up ruled out other causes, the remedy for the problem was absolute total cessation of corticosteroid usage. This article describes the typical history of the problem, the evaluation of these patients, and the distinctive pattern of flaring erythema that ensued when the corticosteroids were ceased. We stress the absolute necessity of total cessation of corticosteroid use as the only treatment for corticosteroid addiction. We also demonstrate that no additional therapy or further consultations were necessary once remission was obtained after topical corticosteroid abuse was halted. PMID- 10459121 TI - Hemorrhoids: what the dermatologist should know. AB - Anorectal disorders are common and more than one half of the population will experience one at some time during their lives. It is important for the clinician to recognize the differences between internal and external hemorrhoids and other anorectal problems such as fissures, abscesses, fistulas, skin tags, and a variety of dermatologic conditions because the treatment is often different. This article will discuss the anatomy, pathophysiology, diagnosis, and treatment of internal and external hemorrhoids. PMID- 10459120 TI - Cutaneous squamous cell carcinomas consistently show histologic evidence of in situ changes: a clinicopathologic correlation. AB - Squamous cell carcinoma on sun-damaged skin is a malignant neoplasm that evolves from its inception as squamous cell carcinoma in situ, which is commonly referred to as an actinic keratosis. In this study, we reviewed 1011 squamous cell carcinomas on sun-damaged skin and found that nearly 100% of these lesions contained histopathologic changes consistent with squamous cell carcinoma in situ at the periphery or within the confines of the squamous cell carcinoma. These malignant changes began in single layer areas of the lower epidermis and evolved into the epidermis and dermis. PMID- 10459122 TI - American Academy of Dermatology 1999 Awards for Young Investigators in Dermatology. Alphavbeta6 integrin mediates latent TGF-beta activation: implications for cutaneous fibrosis. PMID- 10459123 TI - Surgical pearl: A unique surgical marker. PMID- 10459124 TI - Striae formation in two HIV-positive persons receiving protease inhibitors. AB - Steroidal side effects such as "buffalo hump" and visceral abdominal fat accumulation have been reported in association with protease inhibitors. We report two cases of severe disfiguring striae formation in patients with HIV who recently started indinavir therapy. These changes occurred within 3 months of starting treatment. PMID- 10459125 TI - Skin manifestations of a patient with mitochondrial encephalomyopathy with lactic acidosis and strokelike episodes (MELAS syndrome). AB - Skin abnormalities in a patient with newly diagnosed MELAS syndrome are reported. He presented with scaly, pruritic, diffuse erythema with reticular pigmentation on his entire body. Moderate hypertrichosis was also noted on the lower extremities. Thickened arrector muscles with longitudinal and cross sections were seen in the dermis. Analysis of mitochondrial DNA in a biopsy specimen from the biceps brachii disclosed the presence of an A-to-G point mutation at nucleotide pair 3243. Electron microscopy revealed marked increase in the number of morphologically abnormal mitochondria in the arrector muscles of the involved skin. This is the first report of a MELAS syndrome patient with skin manifestations and evidence of morphologically abnormal mitochondria. PMID- 10459126 TI - Adverse cutaneous reactions to ticlopidine in patients with coronary stents. AB - A prospective study was carried out to assess the occurrence and character of adverse cutaneous reactions in patients receiving ticlopidine hydrochloride to prevent subacute thrombosis after having undergone placement of coronary stents. During a 1-year period such patients were requested to report any adverse cutaneous reactions, and those with skin reactions were referred for dermatological evaluation. Among the 136 patients who underwent stent placement by one of the authors, 20 were referred for dermatological evaluation. Of these, 16 (11.8%) fit the case definition of ticlopidine-associated cutaneous reactions. In the first 8 consecutive patients ticlopidine was withdrawn (in 2 of these a rechallenge test was later performed); in the next 8 patients ticlopidine was not discontinued before completion of the intended 4-week period of treatment. Patients remained under weekly follow-up and underwent a weekly blood count. Skin biopsies were obtained in 5 patients with different types of eruptions. The skin reactions appeared from 2 to 21 days after commencement of ticlopidine (mean, 10 days), lasting from 2 to 30 days (mean, 5 days). Only 3 patients had other adverse effects: neutropenia in 1 and abdominal pain and nausea in 2. The most common presentations were urticaria, pruritus, and maculopapular eruption. In 3 patients there were previously unreported reactions: fixed drug eruption, erythromelalgia-like eruption, and erythema multiforme-like eruption. Of note was the rapid clearing of the skin eruption in most cases even when the drug was not withdrawn. It was concluded that adverse cutaneous reactions are relatively common in association with ticlopidine treatment but that serious reactions are rare and the disappearance of the signs and symptoms is rapid, suggesting that discontinuation of the drug is not usually imperative. PMID- 10459127 TI - Metastatic Crohn's disease: remission induced by mesalamine and prednisone. AB - We describe a patient with cutaneous Crohn's disease with extensive cutaneous ulcerations preceding the diagnosis of gastrointestinal Crohn's. The cutaneous lesions responded to mesalamine and prednisone. The patient has been maintained on a regimen of mesalamine alone without recurrence. We discuss available forms of mesalamine and their function in cutaneous disease. PMID- 10459128 TI - Anetoderma of prematurity in association with electrocardiographic electrodes. AB - Anetoderma in premature infants is an uncommon lesion that may be associated with the use of various types of monitoring leads. In 2 infants multiple papules of anetoderma occurred on the forehead in association with the use of gel electrocardiographic electrodes. It is postulated that the cause of these papules was a local hypoxemia caused by pressure from the electrodes. Growth-restricted infants may be particularly predisposed to iatrogenic anetoderma. PMID- 10459129 TI - Proliferative verrucous leukoplakia with cutaneous involvement. AB - Proliferative verrucous leukoplakia (PVL) is a distinct variant of oral leukoplakia characterized by a high rate of malignant transformation. Histologic features are variable and range from epithelial dysplasia to verrucous squamous cell carcinoma. To our knowledge, there have been no previous reports of cutaneous PVL. We present an interesting case of PVL involving the skin. PMID- 10459131 TI - Tadeusz Chorzelski (1928-1999). PMID- 10459130 TI - Improving the safety profile of long-term PUVA therapy. PMID- 10459132 TI - Follicular mucinosis in exaggerated arthropod-bite reactions of patients with chronic lymphocytic leukemia. PMID- 10459133 TI - Lichen amyloidosus: A consequence of scratching? PMID- 10459136 TI - Self-Assessment examination of the American Academy of Dermatology(Identification No. 899-209). AB - Learning objectives: At the conclusion of this self-assessment learning activity, physician participants should be able to assess their own diagnostic and patient management skills with respect to those of their colleagues in the field, use the results of the self-assessment to help determine personal learning needs that can be addressed through subsequent CME involvement, and enhance their ability to comply with the requirements for certification in the specialty of dermatology.Instructions for Category I CME credit appear in the front advertising section. See last page of Contents for page number. INSTRUCTIONS: In answering each question, refer to the specific directions provided. Because it is so often necessary to provide information in questions occurring later in a series that give away answers to earlier questions, please answer the questions in each series in sequence. PMID- 10459134 TI - Role of extracorporeal photochemotherapy alone and in combination with interferon alfa in the treatment of cutaneous T-cell lymphoma. PMID- 10459138 TI - Preface-acitretin for psoriasis therapy PMID- 10459137 TI - Answers to Self-Assessment examination of the American Academy of Dermatology. PMID- 10459139 TI - Retinoids and psoriasis: novel issues in retinoid pharmacology and implications for psoriasis treatment. AB - Oral synthetic retinoids have been established as effective systemic therapy for psoriasis since their introduction for clinical use in the 1970s. Acitretin, the free acid of etretinate and its active metabolite, has replaced etretinate as the retinoid of choice for treating psoriasis because of its more favorable pharmacokinetic profile. Despite the demonstrated clinical success of retinoid therapy in psoriasis and other proliferative skin disorders, their mechanism of action has not been fully elucidated. Altered vitamin A metabolism, characterized by an increase in the formation of retinoic acid, has been demonstrated in psoriatic lesions and is potentially influenced by cytokines such as interferon gamma, which is present in high levels in these lesions. Synthetic retinoids such as acitretin may interfere with such cytokine-induced alterations. Studies on nuclear retinoic acid receptors have shown that acitretin activates all 3 receptor subtypes (RAR-alpha, -beta, and -gamma) without measurable receptor binding; this paradox remains unexplained. Further studies on nuclear receptor binding and activity, including possible receptor crosstalk with vitamin D nuclear receptors, promise to enhance understanding of the usefulness of retinoids in treatment of psoriasis. PMID- 10459140 TI - Acitretin in psoriasis: an overview of adverse effects. AB - Oral retinoids are among the drugs of choice for pustular and erythrodermic psoriasis. In addition, retinoids are effective in combination with other topical and systemic agents for the treatment of plaque-type psoriasis. Acitretin, the active retinoid metabolite, has replaced etretinate in retinoid therapy of psoriasis because of its more favorable pharmacokinetic profile, including a significantly shorter half-life. Retinoids, including acitretin, are potent teratogens, leading to strict requirements for pregnancy prevention during and after their use. Other retinoid side effects are generally preventable or manageable through proper patient selection, dose adjustments, and routine monitoring. Mucocutaneous side effects such as cheilitis and hair loss are the most common dose-dependent side effects, requiring dose reduction in some patients. Less common effects such as hepatotoxicity, serum lipid alterations, pancreatitis, and possible skeletal effects are also discussed. PMID- 10459141 TI - Acitretin: optimal dosing strategies. AB - Acitretin is an oral synthetic retinoid effective in the treatment of psoriasis. As monotherapy, acitretin has been shown to be most effective in treating pustular and erythrodermic types of the disease. Monotherapy with acitretin for plaque-type psoriasis is often less successful; however, its use in combination with other therapies is highly effective in treating this form of the disease. Dose-response studies have established the effective dose range of acitretin as well as the dose-dependence of its side effects. Because both efficacy and side effects can vary substantially among individual patients, proper dosing of acitretin requires a balance between optimizing response and minimizing toxicity for each patient. Optimal dosing for individual patients may be achieved through a dose-escalation strategy involving initiation of therapy at low doses (10 to 25 mg/day) and, if necessary, gradually increasing the dose as tolerated until optimal response is achieved. PMID- 10459142 TI - Acitretin combination therapy. AB - Systemic therapies used in psoriasis such as methotrexate, PUVA, cyclosporine, and acitretin often have side effects that limit their usefulness. Combination therapy, an effective strategy for treating psoriasis, allows use of lower doses of medications to achieve superior therapeutic results with fewer side effects. Acitretin is one of the most popular drugs used in combination with other systemic therapies for psoriasis. Details of the regimens used and results obtained from combining acitretin with PUVA, cyclosporine, and methotrexate are discussed. PMID- 10459143 TI - Acitretin in combination with UVB or PUVA. AB - Combination therapy of psoriasis with acitretin and phototherapy (psoralen ultraviolet A [PUVA] or ultraviolet B [UVB]) offers multiple advantages over use of either modality alone. As monotherapy, acitretin in doses of 50 mg/day is moderately effective, but is associated with numerous side effects. Single modality treatment with UVB or PUVA involves multiple visits over a period of months and is also associated with dose-limiting side effects. When used in combination, lower doses of both modalities can be used more effectively, helping to reduce side effects. In addition, clearing occurs much more quickly, reducing treatment time and number of phototherapy visits. Moreover, patients whose psoriasis does not clear with monotherapy will often achieve significant clearing with the combination of acitretin and phototherapy. PMID- 10459144 TI - Systemic sequential therapy of psoriasis: a new paradigm for improved therapeutic results. AB - Sequential therapy is a treatment strategy involving the use of specific therapeutic agents in a deliberate sequence to optimize the therapeutic outcome. The rationale for this strategy in psoriasis is that it is a chronic disease requiring long-term maintenance therapy as well as quick relief of symptoms and that some therapies available for psoriasis are better suited for rapid clearance while others are more appropriate for long-term maintenance. Sequential therapy involves 3 main steps: (1) the clearing, or "quick-fix" phase; (2) the transitional phase; and (3) the maintenance phase. In the example of sequential systemic therapy described in this article, an acute exacerbation of psoriasis is brought under control promptly with the use of cyclosporine at maximum dermatologic dose (5 mg/kg daily). After 1 month, the transitional phase is initiated with the gradual introduction of acitretin as a maintenance agent. Once the maximum tolerated dose of acitretin has been established, cyclosporine is gradually tapered and acitretin is continued for long-term maintenance with phototherapy (UVB or PUVA) added for improved control if needed. The author proposes sequential therapy with cyclosporine and acitretin as a viable option for patients with psoriasis who require systemic therapy and desire an alternative to methotrexate. PMID- 10459145 TI - Specific in vitro association between the hepatitis C viral genome and core protein. AB - Little is known about the molecular interactions required for hepatitis C virion assembly. The 5' noncoding region (5'NCR) of the RNA genome is highly conserved and has extensive secondary structure. The highly basic core protein is rich in arginine and lysine residues. We postulate that a specific interaction between these structures may be important for virion assembly. Using an RNA gel mobility shift assay, a specific interaction has been demonstrated between the RNA of the 5'NCR and recombinant core protein. Proteins from other regions of the virus do not interact with the viral RNA. The interaction is inhibited competitively by unlabelled sense polarity RNA, but antisense 5'NCR RNA and nonspecific RNAs compete only at much higher concentrations. These data suggest that there is a specific interaction between the 5'NCR of the hepatitis C virus (HCV) genome and HCV core protein. This interaction may be important for the specific encapsidation of the viral genome during HCV replication. PMID- 10459146 TI - Identification of hepatitis C virus seroconversion resulting from nosocomial transmission on a haemodialysis unit: implications for infection control and laboratory screening. AB - Hepatitis C virus (HCV) seroconversion was detected by routine screening in a haemodialysis patient, Patient 1. Serological investigations were undertaken over the following 3 months to determine if further transmission to other patients on the unit had occurred. No additional cases were identified. Twenty-two haemodialysis patients known to have HCV infection were investigated using molecular epidemiological methods to determine if transmission between patients had occurred. HCV viraemia was demonstrated by polymerase chain reaction in 19 of 22 patients (86%). Genotyping showed that eight patients were infected with genotype 1, three with genotype 3 and eight, including Patient 1, with genotype 2. Phylogenetic analysis of viral sequences from the eight patients with genotype 2 revealed three, including Patient 1,with a novel subtype of HCV type 2, and revealed close similarity between viral sequences from patient 1 and those from one other patient, suggesting transmission. This was consistent with haemodialysis histories. Among other patients with genotype 2, there were two with subtype 2a and three others with three separate novel subtypes, as yet undesignated. With the exception of patient 1, all patients infected with novel subtypes were of Afro-Caribbean origin. The HCV prevalence among patients on the haemodialysis unit was high (14%), which may reflect the ethnicity of our haemodialysis population. This case emphasises the risk of nosocomial transmission and the importance of infection control procedures on haemodialysis units, and highlights the usefulness of molecular epidemiological techniques for the investigation of outbreaks of HCV infection. PMID- 10459147 TI - Steatosis and intrahepatic hepatitis C virus in chronic hepatitis. AB - Hepatic steatosis has been reported as one of the characteristics which discriminates hepatitis C from other forms of hepatitis, besides lymphoid follicles and bile duct damage. However, it is unclear whether or not the presence of hepatitis C virus (HCV) itself is associated with the development of steatosis. The possibility that the HCV itself is directly related to the development of steatosis was examined. The intrahepatic core protein levels, as a marker of the HCV load, were correlated with the presence of steatosis in 43 patients with chronic hepatitis C. Among 43 patients studied by Western blotting, the core protein was detected in the liver in 27 (62.8%). On the other hand, hepatic steatosis was observed in 21 (48.8%) of the 43 patients. Importantly, the core protein was detectable in 19 (90.4%) of the 21 patients with steatosis, while it was detected in only 8 (36.4%) of the 22 patients without steatosis (P = 0.008). However, serum HCV-RNA levels as determined by the Amplicor monitor were not significantly different between patients with and without steatosis. Multivariate analysis showed that the serum alanine aminotransferase level (P = 0. 013), body mass index (P = 0.038), and intrahepatic HCV core protein positivity (P = 0.038) were the independent parameters best predictive of steatosis. These results indicate a close relationship between intrahepatic HCV and the development of steatosis, and suggest a possible role of the HCV itself or core protein in the pathogenesis of steatosis in human chronic hepatitis C. PMID- 10459148 TI - Selection of genetic variants of the 5' noncoding region of hepatitis C virus occurs only in patients responding to interferon alpha therapy. AB - Interferon alpha (IFN alpha) can suppress the replication of hepatitis C virus (HCV) in chronically infected patients. However, HCV persists in a significant number of patients despite the normalization of alanine transaminase (ALT) during IFN alpha therapy. In this study, HCV variants in patients under IFN alpha therapy were characterized to examine their role in viral persistence during the therapy. Sixteen patients selected for this study were infected with HCV genotype 1b and remained HCV RNA positive for at least 1 month after onset of therapy. Nine patients responded to the therapy in terms of normalization of ALT (responders), whereas seven patients did not show a significant decrease of ALT level (nonresponders). To examine HCV populations in these patients, the HCV 5' noncoding region (5' NCR) was analyzed by polymerase chain reaction amplification and sequencing. Newly emerging variants of the HCV 5' NCR replaced predominant variants present prior to IFN alpha therapy in six of nine responders. Most predominant HCV variants during IFN alpha therapy carried a nucleotide substitution G to A at nt 231 within the 5' NCR. An analysis of the HCV quasispecies population in one responder revealed that a preexisting variant became predominant under IFN alpha therapy. These results emphasized the importance of the genetic heterogeneity of the HCV genome for viral resistance to IFN alpha. Five of seven HCV isolates from nonresponders were identical to those found in responders with regard to the nucleotide sequence of the 5' NCR. However, no selection of variants of the HCV 5' NCR occurred in nonresponders during the course of therapy. We conclude that IFN alpha treatment leads to the selection of variants of the HCV 5' NCR only in responders and may act differently in nonresponders. Our results suggest that the HCV 5' NCR may be a target of anti-HCV actions of IFN alpha. PMID- 10459149 TI - GBV-C/HGV infection in children with chronic hepatitis C. AB - The role of GB virus-C/hepatitis G virus (GBV-C/HGV), a recently identified member of the Flaviviridae family, in children with liver disease is not well understood. The aims of this study were to evaluate the prevalence of GBV-C/HGV and to clarify its pathogenic role in young patients with chronic hepatitis C. Sixty-four Japanese children and adolescents with chronic hepatitis C virus (HCV) infection, with a mean age of 9.8 years, were evaluated retrospectively. Twenty one (32.8%) of the 64 patients were positive for serum GBV-C/HGV RNA. Only 1 (1.6%) of the 64 patients was positive for antibody against the envelope protein E2 of GBV-C/HGV (anti-E2) and GBV-C/HGV. None of them was positive for anti-E2 alone. There was no significant difference in clinical, virological, or histological characteristics between GBV-C/HGV-positive and GBV-C/HGV-negative patients, except for underlying malignant disease. There was no evidence that GBV C/HGV might affect the response of HCV to interferon therapy in young patients with chronic hepatitis C. The prevalence of GBV-C/HGV infection in young patients with chronic hepatitis C is similar to that in adult patients with chronic hepatitis C, but E2-seroconversion is observed infrequently. Underlying malignant disease is a risk factor for GBV-C/HGV viremia. GBV-C/HGV does not seem to affect the clinical course of young patients with chronic hepatitis C. PMID- 10459150 TI - Infection with GB virus C/hepatitis G virus among blood donors and hemophiliacs in Martinique, a Caribbean island. AB - GB virus C/hepatitis G virus (GBV-C/HGV) RNA was detected by reverse transcription-polymer- ase chain reaction with primers derived from the nonstructural region 3 (NS3) in 9 (4.1%) of 221 blood donors and 2 of 20 (10%) hemophilia patients in Martinique, French West Indies. Anti-E2 antibodies were found in sera from 33 (14.9%) of the blood donors and 5 (25%) of the hemophiliacs. None of the subjects was positive for both GBV-C/HGV RNA and anti E2. Among the 20 hemophiliacs, 12 (60%) had anti-HCV antibodies and 7 (35%) were positive for HCV RNA by PCR. All patients positive for HCV markers belonged to the group of 13 patients exposed previously to blood factor concentrates that were not activated virally. Nucleotide sequences of the 5'-untranslated region (5'UTR) of the GBV-C/HGV genome were obtained for the 10 NS3 PCR positive samples. Phylogenetic comparison of these isolates with reference isolates published previously showed a strong homology with European and American GBV C/HGV strains, 8 isolates belonging to the genotype 2a and 1 isolate to the type 2b. The isolate from 1 blood donor was identified as subtype 1a, indicating the presence of West African type strains. PMID- 10459151 TI - Seroprevalence of antibodies to hepatitis E virus in the normal blood donor population and two aboriginal communities in Malaysia. AB - The prevalence of antibodies to hepatitis E virus (HEV) has been examined in many countries, but such studies have generally been limited to majority populations such as those represented in healthy blood donors or cross sections of urban populations. Due to its major route of enteric transmission, large differences in HEV prevalence might be expected between populations in the same country but with different living conditions. Using an ELISA based on GST-ORF2.1 antigen, the prevalence of IgG-class antibodies to HEV was examined in three distinct populations in Malaysia: the normal (urban) blood donor population and two aboriginal communities located at Betau, Pahang and Parit Tanjung, Perak. IgG anti-HEV was detected in 45 (44%) of 102 samples from Betau and 15 (50%) of 30 samples from Parit Tanjung, compared to only 2 (2%) of 100 normal blood donors. The distribution of sample ELISA reactivities was also consistent with ongoing sporadic infection in the aboriginal communities, while there was no significant relationship between HEV exposure and age, sex, or malaria infection. The high prevalence of antibodies to HEV in the two aboriginal communities indicates that this group of people are at high risk of exposure to HEV compared to the general blood donors, and the results suggest that studies of HEV seroprevalence within countries must take into account the possibility of widely varying infection rates between populations with marked differences in living conditions. PMID- 10459152 TI - Comparative studies on neutralisation of primary HIV-1 isolates by human sera and rabbit anti-V3 peptide sera. AB - IgG binding to V3 peptides and serum neutralising responses were studied in four HIV-1 infected individuals with progressive disease over a period of 31-70 months. The 18-20 mer peptides comprised residues 299-317 (numbering of HIV1 MN) in the N-terminal half of the V3 loop of the envelope glycoprotein gp120 and were derived from the sequences of autologous, as well as heterologous isolates. All four individuals studied lacked anti-V3 IgG binding to at least one autologous V3 sequence. V3 peptides to which autologous sera lacked binding IgG were all immunogenic in rabbits and induced antisera that were broadly cross-reactive by EIA and broadly cross-neutralising to primary HIV-1 isolates. This indicates that the peptides are immunogenic per se and that the respective human hosts have selective defects in recognising the corresponding V3 sequences. Despite the absence of antibody binding to autologous V3 peptides, the human sera had neutralising antibodies to autologous (three out of four cases), as well as heterologous isolates (all cases). Moreover, in vitro exposure of the patients' isolates to autologous neutralising serum or the homologous rabbit antiserum selected for variants with amino acid substitutions close to the crown of the V3 loop or in regions outside the sequence corresponding to peptides used for immunisation. The amino acid exchanges affected V3 positions known to be antigenic and which are also prone to change successively in infected persons. It is likely that neutralising antibodies recognise both linear and conformational epitopes in the V3 loop. Apparently, there are several, but restricted, numbers of ways for this structure to change its conformation and thereby give rise to neutralisation resistant viruses. PMID- 10459153 TI - Sequence requirements for incorporation of human immunodeficiency virus gag-beta galactosidase fusion proteins into virus-like particles. AB - The incorporation of human immunodeficiency virus type 1 (HIV-1) Gag-beta galactosidase (Gag-beta-gal; GBG) fusion proteins into HIV virus-like particles in the presence of HIV Gag proteins was studied. HIV Gag-beta-gal fusion constructs were cotransfected individually into COS7 cells with or without an HIV Gag protein expression plasmid. Release of HIV GBG fusion proteins from the cells were measured by assay of the medium versus intracellular beta-gal activities. Analysis indicates that fusion proteins (constructs HIVGBG, GBG 1919 and 1877) retaining the C-terminal portion of the CA and the adjacent NC domains were efficiently assembled into virus-like particles. Fusion proteins with deleted sequences covering the N-terminal portions of the gag sequences (GBG 831, 1147, 1419, 1447, 1511, 1552, 1600, 1630, 1684, 1715, and 1752) were impaired in entry into virus-like particles. The presence of CA major homology region (MHR) in the fusion proteins had no significant effects on inducing fusion protein incorporation when the C-terminal CA sequences in the fusion proteins were truncated (GBG 1841 and 1801). Subcellular fractionation studies indicated that most fusion proteins including the nonmyristylated one were enriched in the crude membrane fraction. Exceptions to this rule were fusion proteins with intact MHR but truncated C-terminal CA sequences, which possessed low levels of membrane association. However, assembly of fusion proteins into HIV Gag particles did not correlate with their subcellular fractionation or immunofluorescence localization patterns. Overall, the studies suggest that the very C-terminal CA and adjacent NC sequences are the primary determinants for incorporation of HIV Gag-beta-gal fusion proteins into virus particles. PMID- 10459154 TI - Human immunodeficiency viruses type 1 subtypes circulating in Sspain. AB - Genetic subtypes of Human immunodeficiency viruses type 1 (HIV-1) were investigated in 101 HIV-1-infected individuals living in Spain from 1993 to 1998. Samples selected randomly from the HIV clinic population included 29 Spanish native born subjects (28.7%) and 72 foreigners (71.3%). Proviral DNA extracted from peripheral blood mononuclear cells (PBMCs) or viral RNA isolated from plasma was amplified, and endonuclease restriction analysis was carried out on polymerase chain reaction (PCR) products. Restriction fragment length polymorphism (RFLP) analysis on the HIV-1 protease region enabled the characterisation of the different HIV genotypes infecting these individuals. Overall, 38 subjects (37.6%) carried non-B subtypes (A in 26, C in 2, D in 1, E in 2, and F in 7), 31 (81. 6%) of them being immigrants. Direct sequence analysis of PCR products and/or a specific serological assay confirmed the data obtained by RFLP in most individuals tested. In conclusion, different HIV-1 subtypes are circulating currently in Spain, with non-B HIV-1 subtypes being confined mostly to immigrants. PMID- 10459155 TI - Is sexual transmission an important pattern for herpes simplex type 2 virus seroconversion in the Spanish general population? AB - Herpes simplex type 2 (HSV-2) seroprevalence within a community is determined by sexual and perinatal transmission from mother to baby, the two main sources of virus shedding. A seroepidemiological study of HSV-2 was undertaken on a representative sample (n = 3974) of the Spanish population to assess indirectly the relative relevance of these two transmission routes. The sample comprised 1922 men and 2052 women in the age range 5-59 years, stratified by sex and age (5 12, 13-19, 20-29, 30-39, 40-49, and 50-59 years). Sera were screened for HSV-2 specific Ig G antibodies by an enzyme-linked immunoabsorbent assay based on recombinant glycoprotein G2 (gG2). The overall prevalence of antibodies to HSV-2 was 3.6% (95% CI: 3. 1-4.2%). Prevalence by gender did not differ: males (3.6%; 95% CI: 2. 8-4.6%) and females (3.6%; 95% CI: 2.8-4.5%). There were no significant differences between age groups with respect to seropositivity rates. Detection of HSV-2 antibodies was not associated with increasing age, as is expected for a sexually transmitted disease. The fact that seroprevalence rates among the different age groups did not differ suggests that the virus is not circulating in the general population and may be restricted to risk groups only. Similar positivity rates found in the group of females of childbearing age and in the youngest population indicate that perinatal viral shedding is the main source of HSV-2 seroconversion in the Spanish population. PMID- 10459157 TI - Route of TT virus infection in children. AB - TT virus (TTV) is a novel viral agent, detected recently in non-A to E hepatitis cases. Little is known about its natural history or routes of transmission in childhood. For the detection of serum TTV DNA, semi-nested polymerase chain reaction (PCR) was carried out using TTV-specific primers and TTV nucleotide sequences were determined by the dideoxy chain-mediated termination method. Five of the 70 children studied (including 20 hepatitis B virus [HBV] carriers, 40 children born to HBV carrier mothers and 10 children born to hepatitis C virus [HCV] carrier mothers) had serum TTV DNA. Three of the 5 children had siblings (4 in total), so that a total of 9 children were studied to determine the time of initial serum TTV DNA detection. In the 8 seropositive children, the time of serum TTV DNA detection ranged from 6 to 14 months after birth, and TTV DNA persisted thereafter throughout the follow-up period. The TTV DNA-negative child was assessed most recently at 6 months of age. TTV DNA was detected in only 2 of the 4 mothers tested (families 2 and 3). When 271-bp TTV DNA fragments from each of the 8 children were sequenced, the degree of homology between siblings in families 1-3 was 100%, 99.5%, and 92.3%, respectively. The degree of homology between child-mother pairs of families 2 and 3 was 99.5-100% and 62. 6-63.9%, respectively. The distribution of different TTV strains was consistent within families, except for family 3. None of the TTV-infected children had elevated levels of alanine aminotransferase or clinical signs of liver disease. PMID- 10459156 TI - Prevalence of TT virus DNA in eastern Taiwan aborigines. AB - We studied the prevalence of TT virus (TTV) DNA in the general population of the eastern Taiwan aborigine villages, about 11% (34 of 317). There is no association between the presence of HBsAg and TTV DNA or between the presence of HCV RNA and TTV DNA. Therefore, the infection of HBV or HCV and the presence of TTV DNA appear to be independent from each other. The association between the presence of TTV DNA and the elevated alanine aminotransferase (and/or aspartate aminotransferase) activity was also investigated. The presence of TTV DNA was not found to be correlated with abnormal liver function (P = 0.574) when age, gender, and the presence of HBsAg, HCV RNA, and HGV RNA were all considered in the assay. The sequence homology of TTV DNA fragments between different isolates from Taiwan and N22 (the clone obtained from the original prototype strain) from Japan ranged from 84 to 97%. The recombinant protein encoded by the TTV DNA fragment corresponding to the open reading frame of N22 was expressed in E. coli successfully. However, no serum response against the recombinant protein was detected. PMID- 10459158 TI - Genetic characterization of the hemagglutinin of two strains of influenza B virus co-circulated in Taiwan. AB - Two isolates of influenza B virus were obtained in the spring of 1997. One strain, B/Taiwan/21706/97, was isolated from a patient who had acute tonsillitis. The other, B/Taiwan/3143/97, was isolated from a patient who was diagnosed with meningoencephalitis. This implies that the influenza B viruses not only cause respiratory symptoms but may also cause inflammation of the nervous system. Sequence analysis of the hemagglutinin (HA) gene, HA1 domain, indicated that there were remarkable amino acid changes in the strain B/Taiwan/3143/97 compared to B/Victoria/2/87, B/Yamagata/16/88, and B/Taiwan/7/88. The changes in the positions 116, 200, 238, 242, and 271 were correlated with receptor binding. Furthermore, a potential glycosylation site at position 233 was lost. In total, 30 amino acid changes were noted at positions ranging from 116 to 295. These changes may affect the antigenicity of the virus. Phylogenetic analyses also showed that the B/Taiwan/3143/97 was located in an independent lineage, when compared to the reference strains belonging to B/Victoria/2/87 and B/Yamagata/16/88 lineages. This supports the hypothesis that influenza B viruses with distinct genetic characteristic were co-circulated in Taiwan. PMID- 10459159 TI - Comparison of two nested PCR, cell culture, and antigen detection for the diagnosis of upper respiratory tract infections due to influenza viruses. AB - Influenza surveillance requires sensitive and rapid diagnostic methods. Different diagnostic procedures have been evaluated on a selected set of nasal swabs sample collected from patients presenting with acute respiratory infection. One hundred fifty-four samples collected during the peak of the influenza epidemic recorded during winter of 1997-1998 in the south of France were processed for influenza detection using antigen detection (ELISA-immunocapture assay), two different nested RT-PCR assays (targeting M and HA genes), and cell culture. Among 154 samples, 93 (60.4%) were positive for influenza detection. Forty specimens (26%) were positive by ELISA, 77 (50%) by culture, 88 (57.1%) using the multiplex HA PCR and 76 (49.4%) using the M-PCR. Multiplex HA-PCR was thus the most sensitive test. The PCR assay offers an alternative to culture for influenza detection. Nevertheless, culture is efficient for influenza diagnosis and is the only technique that allows the reference centres to collect viral strains and characterise fully new variants. PMID- 10459160 TI - Comparison of respiratory syncytial virus humoral immunity and response to infection in young and elderly adults. AB - Little information about immunity to respiratory syncytial virus (RSV) and disease pathogenesis in elderly persons exists. Humoral immunity to RSV was assessed in 41 young, 56 healthy elderly, and 49 frail elderly adults by measuring baseline RSV specific IgG by enzyme immunoassay (EIA) and microneutralization assay (MNA) in serum. A comparison of the immune response of 11 young and 28 elderly persons with natural RSV infection was also performed. Despite significant differences in age and functional status, no decreases in RSV antibody levels by either EIA or MNA were noted in the elderly compared with the young. Mean baseline MNA titers expressed as log2 were 10.5 +/- 1.1 for the young, 10.5 +/- 1.5 for the healthy elderly, and 10.9 +/- 1.6 for the frail elderly. The frail elderly who attend a daycare had the highest RSV titers to F by EIA at 16.6 +/- 2.0, compared with 15.4 +/- 1.4 and 15.1 +/- 1.4 in the healthy elderly and young, respectively. This finding may reflect recent infection due to their communal setting or increased production of non neutralizing antibody. The immune response of older persons to RSV infection was as vigorous as the younger subjects, with 79% having a >/=fourfold rise in MNA titers compared to 64% in the young. These data suggest that the severe clinical manifestations of RSV in the elderly are not due to a significant defect in humoral immunity. PMID- 10459161 TI - Seroepidemiological study of genogroup I and II calicivirus infections in South and southern Africa. AB - Diarrhoea is associated with the daily death of between 180 and 200 children under the age of 5 years in South Africa. Until recently, many cases and outbreaks of diarrhoea were not associated with a known aetiologic agent. Previous studies using baculovirus-expressed Norwalk virus (NV) and Mexico virus (MxV) capsid antigens have shown that human calicivirus infection is common in South Africa. In this study, our surveillance was extended to different populations, as well as to four other southern African countries: Namibia, Angola, Zimbabwe, and Mozambique. More than 1,700 specimens, some involved in previous cohort studies of infectious diseases, were enrolled in the surveillance. The overall seroprevalence of antibody against NV was >90% for all cohorts except for Mozambican refugees that had 83. 8% sero-positivity. The MxV antibody prevalence was higher than NV, with >95% positivity for all cohorts, except for one in Namibia that had 81% exposure. This study is one of only a few reporting on the concurrent incidence of NV and MxV infections in a cohort study, and has determined that small round structured viruses are prevalent in the local populations of South and Southern Africa. These agents may account for a number of previously unknown or unidentified causes of diarrhoeal illness, in both adults and children, in southern Africa. PMID- 10459162 TI - High prevalence of human papillomavirus type 58 in Chinese women with cervical cancer and precancerous lesions. AB - The prevalence of human papillomavirus (HPV) among 332 Hong Kong Chinese women with abnormal Papanicolaou smears were determined by polymerase chain reaction and restriction fragment length polymorphism analysis. The overall HPV positive rate was 44.3% with 18.6% (16/86) for normal/inflamed cervices, 36.4% (32/88) for condyloma, 64.7% (33/51) for cervical intraepithelial neoplasia grade 1 (CIN 1), 37.9% (11/29) for CIN 2, 68.3 (41/60) for CIN 3, and 77.8% (14/18) for carcinoma. Double HPV infection was detected in 17 of the 147 positive samples, with a significantly higher proportion in patients with normal or inflamed cervices than those with CIN or carcinoma (31.3% vs 10.5%, P =.029). The six most commonly identified genotypes were HPV 16 (33.3%), HPV 58 (23.8%), HPV 11, 18, 31 (8.8% each), and HPV 33 (6.8%). The worldwide uncommon genotype HPV 58 was found to be the second most common genotype detected in patients with cervical carcinoma (6 of 18 patients). HPV 58 infection showed a significant association with CIN/carcinoma (odds ratio [OR] = 3.98; 95% confidence interval [CI] = 1.22-14.35) and a significant trend of increase in prevalence with increasing severity of cervical lesion (chi(2) = 5.84; P =.016). Among Hong Kong Chinese women with abnormal cervical cytology, the detection of HPV 58 carried a positive predictive value of 68.6% for a cervical lesion of CIN 1 or higher severity. The high prevalence of HPV 58 among Chinese women, particularly in patients with carcinoma, has an implication on the design of HPV detection methods and the development of vaccines. PMID- 10459163 TI - Standardization of a PCR-ELISA in serum samples: diagnosis of active parvovirus B19 infection. AB - To standardize a PCR assay for the detection of parvovirus B19 DNA in serum samples three different sample treatments were evaluated on the basis of the efficiency of recovery, reproducibility, convenience of sample handling, and presence of PCR inhibitors. Moreover, the presence of an internal standard competitor as the working reagent at one defined concentration in a competitive PCR-ELISA has been suggested as a valid tool to standardize and validate the assay. The results indicated that serum sample treatment by rapid heating fulfilled the criteria for a routine practice in the diagnostic laboratory. Titration experiments carried out to define the optimal amount of the internal standard competitor to use in PCR-ELISA showed that at 2 x10(2) competitor copies, any amplification interferences between target and competitor sequences were avoided. The internal standard competitor in a competitive PCR-ELISA allows the detection of false-negative results due to PCR inhibitors in the samples or large amounts of target DNA. Heating treatment and competitive PCR-ELISA for the detection of parvovirus B19 DNA were applied to the testing of 347 serum samples, which were submitted to the laboratory for B19 investigation. Of the 34 serum samples that were positive for B19 DNA, 15 were from adult patients and 19 from pediatric subjects. B19 infection was associated with haematological disorders, nonimmunological foetal hydrops, atypical rash, arthropathies, hepatic dysfunction, nonspecific symptoms, and congenital infections. PMID- 10459164 TI - Nucleotide sequence analysis of human T-cell lymphotropic virus type I pX and LTR regions from patients with sicca syndrome. AB - Human T-cell lymphotropic virus type I (HTLV-I) is associated with adult T-cell leukemia (ATL) and tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM). Other inflammatory disorders may occur in HTLV-I-infected patients, such as sicca syndrome resembling Sjogren's syndrome. The sicca syndrome may be the unique clinical manifestation of HTLV-I infection, but is associated frequently with TSP/HAM, which could suggest that sicca syndrome might be an early event in disease progression to TSP/HAM in some cases. We investigated whether peculiar pX and LTR mutations could be related to sicca syndrome, or might argue the existence of clinical progression to TSP/HAM. pX, especially pX(I), pX(II), and pX(IV) ORFs corresponding to Tax cytotoxic T-lymphocyte epitopes, and LTR regions from Caribbean patients who have sicca syndrome with or without TSP/HAM, ATL patients, and healthy carriers were sequenced. The sequences were aligned and compared with ATK-1 prototype and published sequences. LTR sequences exhibited 1.5-2.4% of divergence with ATK-1. pX-sequenced regions showed a lower homology within p12(I) encoding sequences. Only few mutations were found within functionally important regions, but were not associated specifically with the clinical status. Finally, no mutations that could be related to sicca syndrome or argue the existence of clinical progression to TSP/HAM were found. It would be of interest to study the clinical evolution of HTLV-I-sicca syndrome in patients and to determine HTLV-I sequences from peripheral blood and salivary glands at different stages. PMID- 10459165 TI - Analysis of antibody responses against coxsackie virus B4 protein 2C and the diabetes autoantigen GAD(65). AB - Type I diabetes mellitus results from the autoimmune destruction of insulin producing beta cells in the pancreas. Certain viral infections, especially those caused by coxsackie B viruses and related enteroviruses, have been associated with the development of type I diabetes. The sequence homology between the coxsackie B4 virus nonstructural protein 2C (CVB4 p2C) and the major diabetes autoantigen glutamic acid decarboxylase (GAD(65)) provides a basis for the hypothesis of molecular mimicry. In this study, we investigated the prevalence of antibodies directed against nonstructural enterovirus proteins. In addition, a correlation of antibodies against CVB4 p2C and GAD(65) was studied in diabetes patients and in healthy controls. Antibody reactivity against CVB proteins was detected by immunoprecipitation of [(35)S]-methionine-labelled viral proteins and GAD(65) antibodies were measured in a quantitative radio-immunoassay. It was shown that antibodies raised against the nonstructural proteins of CVB4 are very common in the population and a high degree of heterotypic cross-reactivity exists between different enterovirus types. CVB4 p2C-specific antibodies were not only detectable in GAD(65) antibody-positive diabetes patients but also in GAD(65) antibody-negative healthy blood donors. Furthermore, GAD(65) antibodies could not be detected in p2C-positive subjects who had various enterovirus infections, indicating that an antibody response to CVB4 p2C does not necessarily induce a cross-reactive immune response against GAD(65). A correlation was not found between antibodies against GAD(65) and p2C. PMID- 10459166 TI - Localization of the delta-opioid receptor and dopamine transporter in the nucleus accumbens shell: implications for opiate and psychostimulant cross-sensitization. AB - Opiate- and psychostimulant-induced modulation of dopamine transmission in the nucleus accumbens shell (AcbSh) is thought to play a key role in their potent reinforcing and locomotor effects. To investigate the cellular basis for potential functional interactions involving opiates active at the delta-opioid receptor (DOR) and psychostimulants that bind selectively to the dopamine transporter (DAT), we examined the electron microscopic localization of their respective antisera in rat AcbSh. DOR immunoperoxidase labeling was seen primarily, and DAT immunogold particles exclusively, in axon terminals. In these terminals, DOR immunoreactivity was prominently associated with discrete segments of the plasma membrane and the membranes of nearby small synaptic and large dense core vesicles. DAT immunogold particles were almost exclusively distributed along nonsynaptic axonal plasma membranes. Thirty-nine percent DOR-labeled profiles (221/566) either apposed DAT-immunoreactive terminals or also contained DAT. Of these 221 DOR-labeled profiles, 13% were axon terminals containing DAT and 15% were dendritic spines apposed to DAT-immunoreactive terminals. In contrast, 70% were morphologically heterogeneous axon terminals and small axons apposed to DAT immunoreactive terminals. Our results indicate that DOR agonists in the AcbSh can directly modulate the release of dopamine, as well as postsynaptic responses in spiny neurons that receive dopaminergic input, but act principally to control the presynaptic secretion of other neurotransmitters whose release may influence or be influenced by extracellular dopamine. Thus, while opiates and psychostimulants mainly have differential sites of action, cross-sensitization of their addictive properties may occur through common neuronal targets. PMID- 10459167 TI - Gamma-vinyl GABA inhibits methamphetamine, heroin, or ethanol-induced increases in nucleus accumbens dopamine. AB - We examined the acute effect of the irreversible GABA-transaminase inhibitor, gamma-vinyl GABA (GVG, Sabril((R)), Vigabatrin((R))) on increases in nucleus accumbens (NAc) dopamine (DA) following acute administration of methamphetamine, heroin, or ethanol. Methamphetamine (2.5 mg/kg) produced a dose-dependent increase (2, 700%) in NAc DA. GVG preadministration (300 or 600 mg/kg), however, inhibited this response by approximately 39 and 61%, respectively. The lower dose of methamphetamine (1.25 mg/kg), increased DA by 1, 700%. This response was inhibited to a similar extent (44%) regardless of the GVG dose preadministered (300 or 600 mg/kg). In addition, heroin-induced increases in NAc DA (0.5 mg/kg, 170%) were inhibited or completely abolished by GVG (150 or 300 mg/kg, 65 and 100%, respectively). Finally, at half the dose necessary for heroin, GVG (150 mg/kg) also completely abolished ethanol-induced increases in NAc DA following a 0.25 g/kg challenge dose (140%). Taken with our previous findings using nicotine or cocaine as the challenge drug, these results indicate that GVG attenuates increases in NAc DA by a mechanism common to many drugs of abuse. However, it appears unlikely that an acute dose of GVG can completely inhibit increases in NAc DA following challenges with a drug whose mechanism of action is mediated primarily through the DA reuptake site. PMID- 10459168 TI - Non-amine dopamine transporter probe [(3)H]tropoxene distributes to dopamine-rich regions of monkey brain. AB - Drug development in psychopharmacology has adhered to the unwritten precept that compounds targeting monoamine transporters must contain an amine nitrogen in the molecular structure. A series of non-amine-bearing aryloxatropanes that are potent inhibitors of the dopamine transporter (DAT) challenged this precept. In the present study, we investigated the brain distribution of a selective, high affinity DAT non-amine, [(3)H]tropoxene (2-carbomethoxy-3, 4dichloro-3-aryl-8 oxabicyclo[3.2.1] octene), which binds to the DAT in monkey striatum. The autoradiographic distribution of [(3)H]tropoxene was conducted in tissue sections of rhesus (Macaca mulatta) monkey brain. Highest accumulation of the radioligand was detected in the putamen and caudate nucleus, with significant levels also observed in the nucleus accumbens and substantia nigra. Moderate to low levels of [(3)H]tropoxene binding were noted in the hypothalamus, amygdala, ventral tegmental area, and thalamus. The distribution of [(3)H]tropoxene was restricted to brain regions previously identified as expressing DAT, and the relative densities of [(3)H]tropoxene binding sites in various brain regions corresponded to those observed with other selective monoamine radioligands for the DAT. This is the first report to demonstrate that transporter-selective compounds that bear no amine nitrogen in their structure bind selectively to brain regions rich in the transporter. The results support our conclusion that an amine nitrogen is not necessary for compounds to bind to monoamine transporters and distribute in brain according to the known distribution of transporters. The findings provide further incentives to investigate the pharmacological potential of transport inhibitors lacking an amine nitrogen in the molecular structure. PMID- 10459169 TI - Chronic clozapine, but not haloperidol, alters the response of mesoprefrontal dopamine neurons to stress and clozapine challenges in rats. AB - Previously, we demonstrated that serotonin-lesioned rats had an enhanced mesoprefrontal dopaminergic response to restraint stress. This study attempted to extend our knowledge regarding this serotonin/dopamine interaction by seeing if suppression of serotonin metabolism by chronic administration of the atypical antipsychotic, clozapine, would have similar effects. Both typical and atypical neuroleptics require chronic administration in humans before antipsychotic activity is seen. Rats treated for 21 days with clozapine or haloperidol, a typical antipsychotic without significant binding affinity for serotonergic receptors, showed lowered basal dopamine metabolism in the medial prefrontal cortex, the nucleus accumbens, and the striatum, as expected. Basal serotonin metabolism in the prefrontal cortex was also lowered by clozapine treatment, but not haloperidol. One of two challenges were given to chronically treated rats: 30 min of restraint stress or an acute challenge of clozapine. When corrected for baseline differences, both challenges significantly elevated dopamine metabolism in the prefrontal cortex of the clozapine group more than the saline or haloperidol groups. No hyperresponsiveness was seen with serotonin metabolism in the prefrontal cortex or either dopamine or serotonin metabolism in the nucleus accumbens in clozapine-treated, challenged rats. Additionally, this augmentation of the dopaminergic stress response was not seen with a single, acute administration of clozapine. The significance of the clozapine-induced hyperresponsiveness of the mesoprefrontal dopamine system is discussed with regard to clinical efficacy of clozapine. PMID- 10459170 TI - Microdialysis study of the effects of the antiparkinsonian drug budipine on L DOPA-induced release of dopamine and 5-hydroxytryptamine by rat substantia nigra and corpus striatum. AB - The purpose of this study was to determine if systemic treatment with the antiparkinsonian drug budipine was capable of influencing the release of dopamine newly synthesised from L-DOPA in the substantia nigra and corpus striatum of the monoamine-depleted rat. Dual probe microdialysis was therefore employed in freely moving animals pretreated with reserpine (4 mg/kg i.p. 18-20 h earlier) and alpha methyl-p-tyrosine (200 mg/kg i.p. 45 min earlier). Budipine (10 mg/kg i.p.) alone evoked a small but significant increase in basal dopamine efflux in nigra, though not in striatum, but did not affect the spontaneous outputs of DOPAC, 5-HT, or 5 HIAA in either structure. A threshold amount of L-DOPA (25 mg/kg i.p.) stimulated the release of dopamine, DOPAC, and 5-HT (but not 5-HIAA), both in nigra and striatum. The L-DOPA-induced releases of dopamine and DOPAC were greatly accentuated by pretreatment with budipine (10 mg/kg i.p. 45 min earlier), which delayed rather than potentiated the nigral and striatal effluxes of 5-HT. A higher dose of L-DOPA (100 mg/kg i.p.) did not significantly raise the outputs of dopamine or 5-HT, but greatly magnified that of DOPAC. In these experiments, pretreatment with budipine (10 mg/kg i.p.) facilitated the formation of DOPAC from L-DOPA, without increasing the extracellular concentration of dopamine. We conclude from these findings that budipine, at a therapeutically relevant dose, potentiates the release of dopamine newly synthesised from L-DOPA from either end of the nigrostriatal dopamine axis. This effect of budipine could be related to the drug's recently described ability to increase the activity of the converting enzyme, aromatic L-amino acid decarboxylase, and could explain the clinical efficacy of budipine as an adjunct to L-DOPA therapy of Parkinson's disease in man. The significance of 5-HT release to the antiparkinsonian L-DOPA, and the delay in this release caused by budipine, remain to be established. PMID- 10459171 TI - Delta(2)-opioid receptor mediation of morphine-induced CCK release in the frontal cortex of the freely moving rat. AB - Numerous pharmacological data have been accumulated in support of the existence of physiological interactions between cholecystokinin (CCK) and opioids in the central nervous system. With the aim of further characterizing these interactions, an in vivo microdialysis approach was used to directly assess the possible influence of opioids on the extracellular levels of CCK-like material (CCKLM) in the frontal cortex of the awake, freely moving rat. Systemic administration of a high dose of morphine (10 mg/kg i.p.) produced a marked increase (up to +200%) of cortical CCKLM outflow, and this effect could be completely prevented by systemic (1.5 mg/kg i.p.) as well as intracortical (10 microM) administration of the opioid receptor antagonist naloxone. The opioid receptors activated by morphine appeared to be of the delta type because the intracortical infusion of naltrindole (10 microM) also prevented the effect of morphine, whereas CTOP (10 microM), a selective mu-opioid receptor antagonist, and nor-binaltorphimine (10 microM), a selective kappa-opioid receptor antagonist, were inactive. In addition, naltriben (10 microM), which acts selectively at the delta(2) subtype, also abolished the stimulatory effect of morphine on cortical CCKLM outflow, whereas 7-benzylidenenaltrexone (10 microM), a selective delta(1)-opioid receptor antagonist (10 microM), did not alter the morphine effect. Conversely, the direct stimulation of cortical delta(2)-opioid receptors by local infusion of [D-Ala(2)] deltorphin II mimicked the stimulatory effect of systemic morphine on CCKLM outflow. These data indicate that delta(2) opioid receptors play a key role in opioid-CCK interactions in the rat frontal cortex. PMID- 10459172 TI - Distribution of AMPA receptor subunits GluR1-4 in the dorsal vagal complex of the rat: a light and electron microscope immunocytochemical study. AB - The dorsal vagal complex, localized in the dorsomedial medulla, includes the nucleus tractus solitarii (NTS), the dorsal motor nucleus of the vagus nerve (DMN) and the area postrema (AP). The distribution of AMPA-preferring glutamate receptors (AMPA receptors) within this region was investigated using immunohistochemistry and antibodies recognizing either one (GluR1 or GluR4) or two (GluR2 and GluR3) AMPA receptors subunits. The distribution of GluR1 immunoreactivity showed high contrast of staining between strongly and lightly labeled areas. Labeling was intense in the AP and weak in the NTS, except for its medial and dorsalmost parts which exhibited moderate staining. Almost no GluR1 immunoreactivity was found in the DMN. GluR2/3 immunolabeling was present in the entire dorsal vagal complex. This labeling was strong in the AP, the DMN and the medial half of the NTS and moderate in the lateral half of the NTS, except for the interstitial subdivision which exhibited intense staining. Labeling induced by the GluR4 antibody was very weak throughout the dorsal vagal complex. Ultrastructural examination showed that GluR1 and GluR2/3 immunoreactivity was localized in neuronal cell bodies and dendrites. No labeled axon terminal or glial cell body was found. Immunoperoxidase staining in labeled cell bodies and dendrites was associated with intracellular organelles (microtubules, mitochondria, cisternae of the endoplasmic reticulum,.) and/or parts of the plasma membrane. Plasma membrane labeling was often associated with asymmetrical synaptic differentiations. No labeled symmetrical synapse was found using either GluR1 or GluR2/3 antibody. The present results show that AMPA receptors have a widespread distribution in neuronal perikarya and dendrites of the rat dorsal vagal complex. They suggest differences in subunit composition between AMPA receptors localized in the NTS, the DMN and the AP. Ultrastructural data are consistent with the fact that AMPA receptors associated with the plasma membrane are mostly synaptic receptors. However, they also suggest the existence of a large intracellular pool of receptor subunits in neuronal soma and dendrites. PMID- 10459173 TI - Displacement of the binding of 5-HT(1A) receptor ligands to pre- and postsynaptic receptors by (-)pindolol. A comparative study in rodent, primate and human brain. AB - Using receptor autoradiography we examined the displacement of the binding of [(3)H]8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) and [(3)H][N-(2-(4-(2 methoxyphenyl)-1-piperazinyl)ethyl)-N-(2-pyridyl)cy clohexanecarboxamide. 3HCl] (WAY 100635) to 5-HT(1A) receptors by (-)pindolol in the brain of four different species, rat, guinea pig, monkey and human. (-)Pindolol completely displaced the binding of both tritiated ligands at 10(-6) M in all species and regions examined. The affinity of (-)pindolol for presynaptic 5-HT(1A) receptors in the dorsal raphe nucleus was similar to that observed in postsynaptic locations, such as hippocampus (areas CA1, CA3 and dentate gyrus) or entorhinal cortex. Affinity values (K(i)) were in the range 3.8 - 15.9 nM for [(3)H]8-OH-DPAT and 5.8 - 22.3 nM for [(3)H]WAY 100635. In human brain, the K(i) values using [(3)H]8-OH-DPAT as ligand were 10.8 nM in the dorsal raphe nucleus and 6.5 - 13.5 in postsynaptic sites. The present data do not support the hypothesis that (-)pindolol may displace 5-HT(1A) ligands preferentially from presynaptic 5-HT(1A) receptors in the dorsal raphe nucleus, as suggested by electrophysiological evidence. The affinity of (-)pindolol for human 5-HT(1A) receptors is below the mean plasma concentration attained in depressed patients treated with a combination of fluoxetine and pindolol, which indirectly supports an action of pindolol at 5 HT(1A) receptors in these patients. PMID- 10459175 TI - ERRATUM: Everett A.W. 1999. Membrane Recycling Due to Low and High Rates of Nerve Stimulation at Release Sites in the Amphibian (Bufo marinus) Neuromuscular Junction. Synapse 32:110-118. AB - ERRATUM: Everett A.W. 1999. Membrane Recycling Due to Low and High Rates of Nerve Stimulation at Release Sites in the Amphibian (Bufo marinus) Neuromuscular Junction. Synapse 32:110-118. In the above-referenced article, Figure 1 was erroneously reproduced in black and white as converted from a color original, not providing an accurate representation of the staining. The figure is reproduced here as the author intended, from an original black and white rendering. The Publisher sincerely apologizes for this error. PMID- 10459174 TI - Comparative in vivo study of iodine-123-labeled beta-CIT and nor-beta-CIT binding to serotonin transporters in rat brain. AB - Both iodine-123-labeled beta-CIT (2beta-carbomethoxy-3beta-(4-iodophenyl)tropane) and nor-beta-CIT (2beta-carbomethoxy-3beta-(4-iodophenyl)nortropane) have shown to be suitable radioligands for imaging serotonin (5-HT) transporters. [(123)I]nor-beta-CIT has the highest in vitro affinity for 5-HT transporters among beta-CIT analogs reported so far. However, no direct comparison-studies of these two radiotracers as to their in vivo binding to 5-HT transporters have been reported so far. Therefore, it is still unclear which of the two radiotracers is more suitable for single photon emission computed tomography (SPECT) imaging of 5 HT transporters. The purpose of this study was to compare directly in a controlled design the in vivo [(123)I]beta-CIT and [(123)I]nor-beta-CIT binding to 5-HT transporters under the same conditions in rats with the focus on brain kinetic characteristics by means of a two-compartment analysis. We observed that [(123)I]beta-CIT has a higher binding potential and faster kinetics for 5-HT transporters than [(123)I]nor-beta-CIT, suggesting that [(123)I]beta-CIT may be a more suitable radioligand than [(123)I]nor-beta-CIT for imaging 5-HT transporters with SPECT. PMID- 10459176 TI - [Clinical and morphological characteristics and peculiarities of differentiated thyroid gland cancer course]. AB - 221 patients were operated for cancer of the thyroid gland during the last 12 years. Adequate preoperative diagnosis including ultrasound examination, and fine needle aspiration biopsy made it possible to operate 88.7% of patients at earlier (I and II) stages of the disease. Long term results of radical surgical treatment have been studied in 197 (89.1%) patients followed up 5 to 16 years, 49.2% of patients being followed up for no less than 10 years. Majority of patients (95.9%) had differentiated forms of tumors, medullar cancer was detected in 4.1% of patients. Clinical and morphological analysis of differentiated thyroid cancer was carried out in 212 patients. Metastases to lymph nodes of the neck occur more frequently in younger patients, and extracapsular spread of the tumor was revealed in aged persons. Risk factors were male sex, old age, follicular cancer and growth of the tumor through the capsule of the gland. The minimal procedure in case of location of the differentiated tumor in a single lobe of thyroid is extrafascial hemithyreoidectomy with resection of the istmus. Prognosis in radically operated patients is relatively favourable, 5-year survival rate made up 97.5%. PMID- 10459177 TI - [Treatment of peritonitis using ozone and hydropressive technology]. AB - In the experiment on 144 mongrel dogs and 546 white rats the methods of ozone therapy and hydropressive treatment of the abdominal cavity have been worked out. These methods were used in the treatment of 94 patients aged from 13 to 89 years with severe forms of acute peritonitis. The control group consisted of 174 patients with peritonitis. A high antimicrobial effect was detected in the treatment of the abdominal cavity with microdisperse stream of the ozonated solution under pressure. After 1-2 procedures of the intestinal ozone dialysis the number of microbial bodies in the lumen of the small bowel in 65% of patients has decreased 2-3 times, the terms of elimination of the paresis has decreased 3 4 times, and more rapid dynamics of elimination of the endogenous intoxication syndrome was detected. The application of the developed programme of curative measures promote lowering of the lethality from 62.07% in the control group to 37.23%--in the test one. PMID- 10459178 TI - [Combined treatment of acute diffuse peritonitis]. AB - Results of treatment in 199 patients were analysed. The patients were divided into 3 groups. In group 1 (80 patients) acute diffuse peritonitis was treated with conventional methods, in group 2 (62 patients) conventional treatment was supplemented by exchange plasmapheresis, in group 3 (57 patients) conventional treatment was used together with extracorporeal detoxication and elective relaparotomies. Plasmapheresis and elective relaparotomies resulted in increase of effectiveness of the treatment in acute diffuse peritonitis. The intervals between the elective relaparotomies and sessions of plasmapheresis depended on the stage and the course of peritonitis. Lethality rate in conventional method of treatment for acute diffuse peritonitis made up 39%, in plasmapheresis 29%, and in combined treatment--24.5%. PMID- 10459179 TI - [Diagnosis and treatment of tuberculous peritonitis]. AB - Basing on 73 cases, the experience in diagnosis and treatment of tuberculous peritonitis (TP) has been summarized. The analysis of principal clinical symptoms was carried out, diagnostic value of various methods of examination in TP was shown. It was established, that in majority of cases conservative therapy with application of antituberculous preparations contributed to improvement of patients' condition. In 31 patients suppuration of peritoneal liquid as well as bowel obstruction nesessitated urgent surgical intervention. PMID- 10459180 TI - [Surgical treatment of patients with duodenal ulcer]. AB - Ulcer incidence in the Crimeahas increased during the last 9 years from 118.4 to 185.7 cases per 10,000 of adult population, being higher in South Coast and in mountains. From 125 patients with duodenal ulcer after conservative treatment only 12.8% have recovered (follow up for 8-10 years). 29.6% of patients underwent surgery; 1.6%--died without operations of ulcer, 35.2%--needed elective treatment. From 97 patients who died during 4 years of ulcer in Simferopol, 25 (25.8%) were not operated. They were aged and very old patients with long history of the disease. Among 9412 operations for ulcer there were 4.17% of lethal outcomes in perforated ulcer and 11.5%--in bleeding (22.18% being at the peak of hemorrhage); 1.45%--after elective resection of the stomach, 0.79%--after organ saving operations with vagotomy, in SPV--0.56%. According to general hospital surgery data, there were no lethal outcomes after SPV and SPV with pyloroplasty (295 patients). As a conclusion it is stated, that surgery in ulcer should be made earlier, before complications. PMID- 10459181 TI - [High time to make choice: preventive or curative appendectomy?]. AB - The authors advocate prophylactic appendectomy in conditions of wide application of laparoscopic surgery, refer to literature data on the absence of life important functions in appendix and any functional disturbances after appendectomy. Taking into account high rate of development and difficulties in the diagnosis of acute appendicitis, favourable clinical and economical results of laparoscopic appendectomy, the authors suggest wider use of preventive removal of the appendix, especially in definite groups of population. PMID- 10459182 TI - [Foreign bodies of gastrointestinal tract]. AB - 153 patients with swallowed foreign bodies in gastrointestinal tract (GIT) were treated for the last 10 years (1988-1997). In 85 (55.5%) cases foreign bodies (FB) were swallowed by psychopathic persons, in 45 (29.4%)--with the aim of mutilation, in 23 (15.0%)--due to carelessness. In 105 (68.63%) patients the foreign bodies were single, in 48 (31.37%) there were multiple foreign bodies of GIT. In 73 (47.71%) cases FB were found in the stomach. Clinical picture in patients with swallowed FB was variable. Dynamic roentgenological examination together with thorough anamnesis is the most important diagnostic tool in this pathology. The indications for urgent surgical treatment were sticking of the FB, peritonitis, gastro-duodenal bleeding, gastrointestinal obstruction. Elective surgery was used in patients with sticked F.B., with multiple FB, which formed conglomerates, as well as in single FB. with the length over 12-15 cm. Active expectant policy is valid FB of GIT with tendency for passage, due to conservative treatment. PMID- 10459183 TI - [Diagnosis and combined treatment of esophageal chemical burns in children]. AB - The experience of management of 426 children with chemical burns of the esophagus and the stomach has been analyzed. The necessity of earlier (up to 3 hours) lavage not only of the stomach, but also of the esophagus, is substantiated. It is considered advisable to carry out diagnostic fibrogastroesophagoscopy not later than 1 day after the trauma and to give up early prophylactic bouginage of the esophagus. The latfer rather stimulates sear formation than prevents it. The developed scheme of treatment was used in 256 patients with resultant decrease of postburn stenoses of the esophagus 3 times is the results of conventional methods of treatment. PMID- 10459184 TI - [Application of argon coagulator in surgical treatment of ventral hernia]. AB - The results of the application of argon coagulator "Forse-40-GSU-System" in reconstruction of abdominal wall for ventral hernias are presented. The use of this coagulator provides decrease of intraoperative blood loss volume, an achievement of good results during the preparation of a skin flap for autodermoplasty. PMID- 10459185 TI - [Use of leukinferon for prophylaxis of pyogenic complications in victims with thoracic wounds]. AB - The results of treatment of 131 victims with penetrating wounds of the thorax complicated by profuse bleeding, are presented. 36 of them after surgery to prevent pyogenous complications received 5 injections of leukinferon (test group); 95 patients who were not treated by leukinferon made up control group comparable with the test group by sex, age and character of wounds. Comparative analysis of the results showed that treatment with leukinferon has resulted in decrease of postoperative complications. In the test group the number of complications made up 47.2%, in control group--61%. Al the same time in the former group there was only one pyogenous complication (2.9%), while in control group--10 (10.6%). After the course of leukinferon leucocytic formula of the blood has become normal, the number of patients with immunodeficiency and imbalance of proteins markers of the acuteness of inflammatory process has decreased. PMID- 10459186 TI - [Kallikrein-kinin system in acute empyema of pleura and pyopneumothorax]. AB - In 56 patients with acute empyema of the pleura the level of aggregate kininogen and prekallikrein, free kallikrein and kininase activities in blood plasma and supernatant of pleural exudate were studied. It was established that epsilon aminocapronic acid, contrykal and gordox suppress free kallikrein activity in the blood plasma and supernatant of pleural exudate. Intravenous and local application of proteolysis inhibitors resulted in improvement of the outcomes of the disease. PMID- 10459187 TI - [Diagnosis and treatment of diaphragm traumatic ruptures]. AB - 29 patients with traumatic ruptures of left cupola of the diaphragm were studied. Pain, dispnoe, tachycardia more intensive after a meal due to repletion and dislocation of the stomach into pleural cavity and its pressure on the organs of the mediastinum were the main symptoms of the disease. The diagnosis of traumatic rupture of the diaphragm was made on the base of clinical and roentgenological examination. In the acute period of trauma the diagnosis is difficult, especially in combined trauma. In 92.3% of cases the stomach and the bowel move into the pleural cavity, which may simulate pneumothorax or relaxation of the diaphragm. Contrast examination of the gastro-intestinal tract and pleural puncture help in differential diagnosis. All the cases of traumatic ruptures of the diaphragm should be subjected to surgical treatment. Urgent operation is indicated in continuing bleeding, incarceration of organs, acute cardiopulmonary insufficiency due to the collapse of the lung and dislocation of mediastinal organs of into contralateral position. Thoracotomy in VII intercostal space is considered as a valid approach. There were 2 (6.9%) lethal outcomes after 29 operations. Long term results are quite favourable. PMID- 10459188 TI - [Prognosis of postinjection abscesses course with use of mathematical model]. AB - Virulent and persistent characteristics of staphylococci isolated from 100 patients with various types of course of the abscessed forms of postinjection suppuration were defined. Comparative analysis of 11 biological characteristics of the microorganisms was carried out. An important role of the complex of properties of Staphylococcus aureus was established, including lysozyme, proteolytic, fibrinolytic activities, ability for inactivation of immunoglobulins through their Fc-receptors' binding to protein A, antilysozyme, "antiinterferon", anticomplement activities in determination of protracted character of festering and inframmatory process initiated by them. The model for prognosis of the course of postinjection abscesses was developed with the help of discriminant analysis, being based on the analysis of the informative properties of the pathogen. PMID- 10459189 TI - [Surgical treatment of pyo-necrotic forms of diabetic foot]. AB - The authors present the experience of combined surgical treatment of 94 patients with complicated "diabetic foot". In the majority of patients (92.8%) there was ischemic form of diabetic foot. Wide usage of staged necrectomies as treatment modalities, preferable use of economical amputations and exarticulations, continuous intensive intraarterial infusion (heparine, desaggregants, antibiotics, insulin suspension) contributed to preservation of the limbs in 82.6% of cases. PMID- 10459190 TI - [Anaerobic paraproctitis complicated by phlegmon of left gluteal region, thigh and shin]. PMID- 10459191 TI - [Surgical treatment of epiphrenical diverticulum and stomach cancer]. PMID- 10459192 TI - [Method of surgical treatment of inguinal hernia]. PMID- 10459193 TI - [Buttock injury penetrating into abdominal cavity]. PMID- 10459194 TI - [Pelvic evisceration in locally advanced cancer of rectum with the ingrowth into urinary bladder]. PMID- 10459195 TI - [Transjugular intrahepatic portosystemic shunt in treatment of portal hypertension]. PMID- 10459196 TI - [Leukemias--diseases of haemopoietic organs stroma (hypothesis)]. AB - Like bones and other tissues in the organism, hemopoietic tissue is a constantly regenereting system in which stem hemopoietic cells (SHC) are involved in mitosis like stem cells of other tissues, this suggesting a universal mechanism of cell repopulation. Hemopoietic connective tissue is a special structure for autoregeneration and differentiation of stem cells. It forms hemopoiesis as the environment in which cells develop. Cell repopulation is the function of connective tissue and bone marrow. In leukemia involvement of the stroma creates conditions for tumor cell growth. Connective tissue disease at first manifests by imbalanced cell-to-cell interactions and then disorders cell repopulation and leads to appearance of abnormal cell forms. Abnormalities in cell development involve alteration of their biological properties. Fibrosis is a common biological phenomenon. It is one of manifestations of connective tissue functions. Fibrous tissue overgrowth in leukemia is to be regarded from a general biological viewpoint. Normally extension of the function of bone marrow hemopoiesis involves or is parallel with connective tissue growth which does not impair balanced interactions between cellular structures, their regeneration and degradation. Myelofibrosis, a form of fibrosis manifestations, is observed (in different measure) in all leukemias and is their integral component. Hemopoietic connective tissue is an "incubator" of SHC, and therefore, development of fibrosis is determined by the initial connective tissue involvement (disease) of nontumorous origin. In leukemia the same biological regularities, maintaining the organism viability, are in force, but their effect is distorted, because in leukemia the bulk of cells is many times higher than in health or even in disease. Moreover, increase of the bulk of cells occurrs under conditions of deficient bone marrow hemopoiesis, present to this or that measure in leukemia. PMID- 10459197 TI - [Study of erythrocytes by atomic force microscopy]. AB - Working principle of Nanotop-202 atomic force microscope is described. The morphology of peripheral blood erythrocytes is studied under this microscope. A method for preparing the specimens is developed and the resultant cell images are described. The results demonstrate the potentialities of the method for morphometry, studies of fine structure of membrane surface, and manipulations with erythrocytes. Prospects of atomic force microscopy in hematological studies are discussed. PMID- 10459198 TI - [Laboratory monitoring of distraction bone formation]. AB - The course of the early period of bone formation during distraction osteosynthesis can be safely, rapidly, and objectively monitored and predicted by hematological and immunological parameters (lymphocytes, rosette-forming cells, monocytes, immunoglobulin A, heterophilic hemagglutinins), which are tested before and after surgery on days 1 and 7-10 and on day 10 of distraction. PMID- 10459199 TI - [Test for detection of activated partial thromboplastin time using ellagic acid]. AB - A simple and sensitive method for estimation of activated partial thromboplastin time (APTT) is developed, making use a complex reagent containing the activator (plant phospholipids) and contact factor (ellagic acid). The test requires additionally only 0.025 M CaCl2. The test is more sensitive to the presence of heparin in the blood and to insufficiency of blood clotting factors VIII and IX than the reagents containing insoluble substances (kaolin and animal phosphatides). Addition of soluble ellagic acid into reagent for APTT estimation allows studies on optic coagulometers. PMID- 10459200 TI - [A method for studying intravascular platelet aggregation in vitro]. AB - A simple available method for evaluating intravascular platelet aggregation is proposed. It consists in graphic recording of disaggregation of platelet-rich citrate plasma, which indicates the degree of intravascular aggregation. Intravascular aggregation is notably increased in coronary patients and negligible in normal subjects. The method may be used for the diagnosis of diseases with a high thrombogenic risk. PMID- 10459201 TI - [Improving the efficacy of tuberculosis immunodiagnosis using anti-human IgG monoclonal antibodies]. AB - Monoclonal antibodies (MAb) F5 to human IgG were used for creating immunoperoxidase conjugate. MAb dissociation constant was 10(-9)M-1 and the number of binding sites 1. Enzyme immunoassay (EIA) and immunoblotting showed that MAb F5 specifically recognize conformation epitope on intact human IgG molecule but not on other human immunoglobulins or denatured IgG. The resultant peroxidase conjugate with MAb F5 was used for EIA titration of antibacterial antibodies in sera from 30 patients with pulmonary tuberculosis, 28 patients with nonspecific pulmonary diseases (bronchitis and/or asthma, pneumonia), and 12 donors. For comparison similar studies were carried out with reference commercial immunoperoxidase conjugate to human IgG(H + L) manufactured at N. F. Gamaleya Institute of Epidemiology and Microbiology. Mycobacterium tuberculosis monoantigen (mol. weight 15-18 kD), affinity isolated by antibacterial MAb S4C1G4 (alpha S4C1G4), and PPD (Batch RT 45, Stattens Seruminstitut, Denmark) were used. Sensitivity and specificity of serum antibacterial antibodies were compared. The specificity of conjugate based on MAb F5 with monoantigen alpha S4C1G4 was 78.21%, sensitivity 94.50%, while those of conjugate to human IgG(H + L) were 53.30 and 76.89%, respectively (p < 0.001). For PPD the specificity and sensitivity were 56.75 and 72.33%, respectively (conjugate with MAb F5) versus 47.67 and 62.38% for conjugate against IgG(H + L), p < 0.001. Similar values were obtained in assessment of the concentrations of antibodies to alpha S4C1G4 for MAb F5 conjugate: specificity and sensitivity 47.16 and 71.56%, respectively, versus 23.67 and 95.16% for PPD (p < 0.05). No statistically significant differences between the experimental groups were detected with IgG(H + L) conjugate. We believe that specific MAb-based conjugate to human IgG will improve the efficacy of EIA as a method for the diagnosis of pulmonary tuberculosis. PMID- 10459202 TI - [CD-markers in the practice of clinical diagnostic laboratories (Lecture)]. PMID- 10459203 TI - [Activation of lymphocyte nucleoli in culture adding the putative allergen to culture media: a tool for allergy diagnosis]. AB - A method for diagnosis of allergy from activation of the peripheral blood lymphocyte nucleoli in short-lived cultures of peripheral blood leukocytes of 16 patients with clinical manifestations of drug allergy is described. The tests were positive in 75% cases. PMID- 10459204 TI - [Clinical application of C-reactive protein ("NicoCard"--a new method for traditional test]. AB - C-reactive protein (CRP) is a unique marker of acute phase response to inflammation. Measurements of CRP in the blood are widely used for diagnosis and monitoring in infections and diseases, for evaluating the activity of inflammation, choice of adequate therapy, control and prediction of disease course. Fulminant course of many diseases dictates the necessity of rapid easily interpreted quantitative non-instrumental test fit for manipulations with whole blood. NicoCard CRP kit manufactured by the Nicomed Firm allows a rapid (2 min) and accurate (quantitative using NicoCard Readers) detection of inflammation and evaluation of its severity, helps differentiate between bacterial and viral infections, choose adequate therapy (antibiotics, steroids, antiinflammatory agents), and monitor the treatment efficacy. PMID- 10459205 TI - [Medium molecular weight molecules and their fractions in Astrakhan rickettsial fever]. AB - The level of medium-weight molecules (MWM) over the course of disease and changes in the lipid share of MWM fractions at the peak of disease were studied in patients with Astrakhan rickettsial fever (ARF). The level of MWM was significantly increased in comparison with the control and MWM concentration clearly correlated with the severity of intoxication. A level higher than 600 U is an unfavorable sign of a grave course. The significance of the lipid part of MWM fraction in the formation of the intoxication syndrome in ARF is shown. PMID- 10459206 TI - [Transferase activities in the cerebrospinal fluid of patients with craniocerebral injury]. PMID- 10459207 TI - [Deontology in bacteriology]. PMID- 10459208 TI - [Trend in rapid diagnosis of purulent infections pathogens and rapid assessment of treatment efficacy]. AB - A method for rapid diagnosis of wound infection based on gas chromatographic detection of metabolites of anaerobic bacteria is proposed. If acetic acid is present in pathological material from the wounds, bacteriological analysis detects facultative anaerobic bacteria in 71% cases. PMID- 10459210 TI - [Hemodynamic stroke]. PMID- 10459211 TI - [Benign hyperbilirubinemias]. PMID- 10459212 TI - [Mechanisms of inflammation and immunity regulation in the pathogenesis of Lyme disease]. PMID- 10459213 TI - [Clinical efficiency of infrared impulse laser therapy in patients with rheumatoid arthritis]. PMID- 10459214 TI - [The comparative assessment of follow-up for hypertensive patients]. AB - The quality of outpatient follow-up was compared in hypertensive patients of the primary care system in 1995 vs 1997. Case records have been retrospectively analyzed for 1301 patients and 1335 ones treated in 6 Kemerovo outpatient clinics in 1995 and 1997, respectively. A special sheet was used for recording the data. More investigations of the target organs were made in 1997. An increase was found in the number of tests for total cholesterol, creatinine, blood glucose, blood count, urinalysis, ultrasound scanning of the kidneys and adrenals, ECG. More patients were taking drugs. Adelphan administration decreased by 32%, clophelin by 30%. More frequently were prescribed ACE inhibitors, calcium antagonists, beta adrenoblockers. Quality of care for hypertensive patients is still low. This is explained by deficient social care, poor medical education of the patients, rejection of active treatment by many patients. PMID- 10459215 TI - [Effectiveness of enap and oliphen combined treatment of chronic pulmonary hypertension]. PMID- 10459216 TI - [Aspects of hypertensive heart in arterial hypertension and insulin-resistant syndrome]. AB - 61 patients with essential hypertension stage II were examined using echocardiography, Holter ECG monitoring, assessment of carbohydrate and lipid metabolism, hormonal profile of arterial hypertension (AH). AH criterium associated with insulin resistance was combination of AH with at least one of the components of the "death four": obesity, hypertriglyceridemia diabetes mellitus or impaired glucose tolerance. Hypertensive heart in AH associated with insulin resistance is severe left ventricular hypertrophy, diastolic myocardial dysfunction. The above disturbances appear prior to elevated blood levels of insulin and aldosteron suggesting the role of the hereditary factors. In hypertensive heart in the syndrome at late stages paroxysmal fibrillation is seen. Episodes of coronary events and ventricular arrhythmia arose irrespective of insulin resistance syndrome. PMID- 10459217 TI - [Interferon therapy in chronic hepatitis C and cellular immunity]. PMID- 10459218 TI - [Comparative analysis of existing criteria in the assessment of iodine-deficiency in dwellers of the Far North areas]. AB - New standards of the thyroid volume proposed by F. Delange (1977) are now being introduced. In view of this, we compared incidence of goiter by F. Delange and by R. Gutekunst (1988) criteria. The examination covered 3920 schoolchildren in 12 settlements of the Tyumen region in 1996. Iodine deficiency was mild to moderate. The greatest deficiency was observed in the North. Ultrasonic investigation of the thyroid by standards of R. Gutekunst (1988) was also indicative of the greatest iodine deficiency in the North. In the Far North, Polar Urals there was severe goiter endemia, in the rest areas the endemy was moderate. By F. Delange (1977) criteria, mild and moderate endemy was in the Far North and Polar Urals, the endemy was absent in the rest areas. As shown by comparison of the palpation data to thyroid ultrasonography, iodinuria, R. Gutekunst criteria are more reliable whereas criteria of F. Delange provide overestimated data. PMID- 10459219 TI - [Effectiveness of grandaxin in the treatment of somatoform disorders]. PMID- 10459221 TI - [The evaluation of the effectiveness and tolerance of movalis in patients with osteoarthrosis]. PMID- 10459220 TI - [Berodual: clinical experience]. PMID- 10459222 TI - [On the ethics of clinical trials of new drugs]. PMID- 10459223 TI - [Possibilities for the prognosis of cardiac rupture in patients with acute myocardial infarct]. PMID- 10459224 TI - [Thrombocytopenias]. PMID- 10459225 TI - [Adolf Kussmaul (1822-1902) as a doctor, scientist, teacher]. PMID- 10459226 TI - [Memorable dates in the history of medicine in 1999]. PMID- 10459228 TI - [Physicians' position towards the Ansvarsnamnden should be stronger]. PMID- 10459227 TI - [If we cease to vaccinate the diseases will come back]. PMID- 10459229 TI - [We are not talking about the same patient group when it comes to Dormicum]. PMID- 10459230 TI - [Increase support to patients with Rett syndrome!]. PMID- 10459231 TI - [More about vitamin AD and sudden infant death]. PMID- 10459232 TI - [Caution for the Respiration Guard!]. PMID- 10459233 TI - [The smoking personnel is only a part of the necessary measures]. PMID- 10459234 TI - [Does physicians' learning capacity cease in connection with retirement?]. PMID- 10459235 TI - [A new vaccination campaign for better protection against whooping cough of infants]. PMID- 10459236 TI - [Special needs of adults with neuropsychiatric disabilities]. PMID- 10459237 TI - [Statistical quality indicators in health care--there is a lot to do]. PMID- 10459238 TI - [Cystic fibrosis--an ancient disease with renewed topicality. History and clinical practice]. AB - Cystic fibrosis (CF) is a general exocrinopathy chiefly characterised by airway and nutritional symptoms, though several other organ systems may also be affected. Intensive collaboration between clinicians and research scientists has enabled great advances to be made in our understanding and treatment of the disease. New mucolytic treatment, new approaches to combating infections, and improvements in nutrition have already appeared. Just around the corner await gene therapy and new drugs impinging directly on the mechanisms whereby the gene mutations exert their harmful effects. The median life expectancy of CF patients is now over 40 years. CF is currently the concern of multiple disciplines. PMID- 10459239 TI - [Adverse effects of dental materials. Important that also physicians report]. PMID- 10459240 TI - [Nordic unity on DAMP/ADHD. A current document summarizes the current knowledge]. PMID- 10459241 TI - [Children of letters--ghosts from the past with a new name. A small group if stringent criteria are used]. AB - Hyperkinetic disorders, MBD (minimal brain dysfunction), DAMP [deficits in attention, motor function (or muscular control) and perception], ADD (attention deficit disorder), and ADHD (attention deficit hyperactivity disorder), are currently the focus of interest among parents, clinicians and researchers in Sweden; and guidelines for the assessment and treatment of such disorders were published in the USA in 1997, and in Europe in 1998. However, despite the accumulated knowledge, and new hypotheses which have been proposed, e.g., that ADHD is "a disorder of adaptation," there is no consensus as to the understanding, treatment and prevention of these disorders. In a consensus statement published by an NIH (National Institutes of Health) panel in the USA it was concluded that, after years of clinical research and experience, our knowledge of the aetiology of ADHD remains speculative, and no documented strategies for its prevention are available. A review of Swedish views and concepts of these disorders since 1950 showed discussion to have been characterised by more similarities than differences, and that nothing really new had emerged. However, differences have existed in the sphere of general education. Since WWI, changes in the organisation of the statutory school system have been designed to improve both education and health among schoolchildren. Such changes seem to have had both beneficial and adverse effects on the overall health of the children. The prevalence of ADHD-like problems declined during the period, 1949-70, when all Swedish six-year-olds were screened for school readiness with a standardised national test, and the class in which a child started school was dependent on intellectual capacity, overall mental age, and the presence or absence of reading, spelling and learning difficulties, and of behavioural problems. Since this was discontinued after reorganisation of the school system in 1970, the prevalence of problematic behaviour has once again increased. Thus, in the search for new approaches to the support of children with hyperactivity and attention deficit problems, analysis of the organisation of the school system should not be forgotten. PMID- 10459242 TI - [National registries for shoulder and elbow arthroplasties are established]. PMID- 10459243 TI - [Oats can be included in gluten-free diet]. AB - In the management of coeliac disease, it has been widely accepted that oats must also be excluded from the diet, along with wheat, rye and barley. The article consists in a review of published reports, and an account of our experience of including oats in the gluten-free diets of adults. Oats were found to be safe and well tolerated by adults with coeliac disease and dermatitis herpetiformis, though the risk of wheat contamination of commercial oat products remains a cause of concern. Similar findings were reported from a study of adolescents, but no such studies have been made of small children. Thus, the inclusion of oats, known to be a fibre-rich, naturally gluten-free food, would broaden the range of foodstuffs tolerable to coeliac patients, though for safety reasons they should be used only by adults until more information is available. PMID- 10459244 TI - [A case report. A vaccine against tick-borne encephalitis caused polyneuritis]. PMID- 10459245 TI - [Prescribed exercise can help the patient to change his habits]. PMID- 10459246 TI - [A medical historical pause. The swan in Lund sculptured in a satin-soft walnut]. PMID- 10459247 TI - [Social aspects of the adaptation and human ecology]. AB - Influence of natural factors on humans or human migration into extreme conditions alters relationships between population and environment, therefore functional resources are mobilized. This alteration sometimes induces genotypic and phenotypic inadequacy to new, more strict requirements of natural and occupational factors. That is because evolution develops and genetically fixes in most people various mechanisms providing and supporting biologic balance. Requirements set in favorable environmental conditions for homeostatic systems including immune one are adequate to genotype of most individuals. When those mechanisms are not fixed and/or environmental conditions appear unfavorable the changes occur that could transform into disease. PMID- 10459248 TI - [Intellectual occupations and chronic stress in shift work]. AB - Genesis of chronic occupational stress covers steps of sequent transformation from functional state to strain, fatigue and overfatigue. Physiologic features of these conditions were analyzed, so the conditions could be assessed through functional levels, relationships between central nervous and cardiovascular systems and inner links of these systems, coordination of diurnal rhythms. Workers having higher occupational psychoemotional strain, shift work schedule and 12-hours working day are significantly prone to neurotic disorders. PMID- 10459249 TI - [Prophylaxis of muscular stress in physical labor]. AB - The authors demonstrated that work of female molders, acquisitors and stacking packers is characterized by physical load, great number of cliched manual movements, intensive inclinations of corpse and constrained working posture. Prolonged exposure to unfavorable occupational factors results in neuromuscular and cardiovascular stress that induces pathologic changes with increasing length of service. Complex of prophylactic measures suggested by the authors proves effective as releases working strain and delays fatigue during the working shift. PMID- 10459250 TI - [Studying strain and recovery of initial cardiovascular functional state in accordance with "vegetative tone background"]. AB - The aim of the investigation is the observation of strain during distinct physical and psychic load and particularly the recovery of the functional resting state of the cardiovascular system during restitution after load. Parameters of performance did not differ significantly between vagotonics and sympathotonics. Subjective appraisal of the ability to recover and relax after load comes off better by vagotonics than by sympathotonics. Vagotonics show a greater variability of parameters of the heart rhythm during transitional stages than sympathotonics do. This is a sign of good regulation of the tone by vagotonics during adaptation of the body to a new situation. We conclude a better come off of restitution of the functional state of the body after load, a better ability to recover and a better ability to hold out against new load. PMID- 10459251 TI - [Specific aspects of fatigue formation in professionals working with video displaying terminals]. AB - The article presents laboratory results describing influence of hand and arm temperature on capacity of fingers' flexor muscles for dynamic physical work. New means and method for individual protection from local vibration are designed. The authors discuss efficiency of these novelties for exposure of hands to local physical load and vibration. PMID- 10459252 TI - [The influence of occupational factors on clinical and physiological states of those working at video-display terminals]. AB - The studies proved users of videodisplays to be under exposure to occupational hazards cumulative even in total 4-hour daily work at the videodisplay. That induces energetic and informational stress that clinically resembles fatigue neurosis. The examinees demonstrated premorbid condition, lower physiologic resources, homeostatic potential and occupational health quality. PMID- 10459253 TI - [Occupational aspects of work conditions and assessment of several clinical biochemical and chemiluminescence parameters in workers of oil processing industry]. AB - The authors present materials concerning hygienic evaluation of work conditions and health state in individuals exposed to pyromellitic dianhydride and other aromatic substances detected in the air of workplace. Changes in lipid peroxidation were evaluated through serum and urine chemiluminescence levels, shift of energy metabolism parameters was assessed via dynamics of ATP, ADP, AMP and adenyl system enzymes in RBC. Examinees of groups 2 and 3 demonstrated significant changes in adenyl system parameters of RBC and in serum and urine chemiluminescence. Conclusion is that serum and urine studies using bioenergy and chemiluminescence methods help to evaluate compensatory and metabolic resources, to reveal premorbid condition and to correct it without transitory disablement of the subjects. PMID- 10459254 TI - [Occupational psychiatry: major stages and prospects of development (review of literature)]. PMID- 10459255 TI - [Work regulations in occupational prophylaxis (territorial center)]. AB - Activities of therapists working in occupational service centers are characterized by significant peculiarities mostly connected with occupational medical inspection. Given high social value of the expertise, specification of scientifically justified working load for those therapists working in various occupational service centers becomes especially topical. PMID- 10459256 TI - ["Operational" and "trace" physiological parameters in hard labor]. PMID- 10459257 TI - [On osteo-muscular disorders in female workers of sewing factories]. PMID- 10459258 TI - [Management process in expedition shift rotation work]. PMID- 10459259 TI - International medical readiness. PMID- 10459260 TI - Disease prevention while deployed. PMID- 10459261 TI - Environmental air sampling to detect biological warfare agents. AB - The rapid and unequivocal detection and identification of biological warfare agents is a major goal of military and civilian defense authorities. To identify agents of concern in an environmental sample, a reliable, region-specific characterization of the microorganisms found naturally at the sampling location is required. We have analyzed environmental air samples from Korea, Kuwait, and Bahrain by polymerase chain reaction and temporal temperature gradient electrophoresis and have produced genetic fingerprints of the natural microbial flora in these regions. Results are displayed as specific bar code patterns against which the unique patterns of potential biological warfare agents appearing in a sample can be quickly discriminated. Data are stored on compact disk, along with other laboratory analyses and relevant meteorological data, and are available to appropriate authorities and researchers. PMID- 10459262 TI - Near-field speech intelligibility in chemical-biological warfare masks. AB - It is common knowledge among field personnel that poor speech intelligibility can occur when chemical-biological warfare (CBW) masks are worn: indeed, many users resort to hand signals for person-to-person communicative purposes. This study was conducted in an effort to generate basic information about the problem; its focus was on the assessment of, and comparisons among, the communicative efficiency of seven different CBW units. Near-field word intelligibility was assessed by use of rhyming minimal contrast tests; user and acoustic restrictions were studied by means of diadochokinetic tests and system frequency response. The near-field word intelligibility of six American-designed masks varied somewhat, but overall it was reasonably good; however, a Russian unit did not perform well. Second, three of the U.S. masks were found to produce less physiological restraint than the others, and the Soviet mask produced the greatest physiological restraint. Finally, a few of the CBW masks also exhibited very low levels of acoustic distortion. Accordingly, it was concluded that two of the several configurations studied exhibited superior features. Other factors being equal, they can be recommended for field use and as a basis for the development of future generations of CBW masks. However, it also should be noted that although these devices provided reasonably good speech intelligibility when the listener was close to the talker, they do not appear to do so even at minimal distances. PMID- 10459263 TI - Suicide in the U.S. Marine Corps, 1990 to 1996. AB - Epidemiologic studies of suicide in the military have not controlled for the higher suicide rates of the unemployed expected in comparative national populations. This study compared the observed number of suicides among U.S. Marine Corps personnel from 1990 to 1996 with the expected number based on rates for the employed general U.S. population. Standardized mortality ratios were calculated to identify demographic groups with higher or lower than expected numbers of suicides. The scan statistic and the Knox technique were used to evaluate potential suicide cluster patterns. Overall, there were fewer suicides in the Marine Corps (n = 213) than expected (n = 225). Hispanic and other ethnic group males and female Marines had greater than expected numbers. Evidence for suicide clustering in time and space was equivocal. PMID- 10459264 TI - The implications of protocol-based care on dental services in the military. AB - Downsizing, limited resources, and increasing costs provide challenges to the military health system. Variations in the diagnosis and treatment of dental disease add to the demands on the delivery system to provide access and ensure quality for uniformed personnel. Evidence-based dentistry is the conscientious, explicit, and judicious use of current best evidence in making decisions about the care of individual patients. An evidence-based practice combines individual clinical expertise with the best external evidence available from systematic review of research findings. It provides a scientific basis for patient care, planning and implementation of health services, and development of health policy. Practice guidelines formulated on scientific evidence can reduce variations in the diagnosis and treatment of various dental conditions. A risk assessment protocol for treating dental caries can reduce operative dental treatment recommended at the initial examination and decrease the need for restorative care during a military career. PMID- 10459265 TI - The prevalence and treatment of dental caries among Israeli permanent force military personnel. AB - A survey was conducted to determine dental caries prevalence and treatment among 1,095 25- to 44-year-old permanent force Israeli military personnel. Caries experience, by decayed, missing, and filled permanent teeth (DMFT), was 11.66, with an average of 1.37 untreated caries, 2.40 extracted teeth, and 7.90 treated teeth. Caries was positively associated with age (p < 0.001). Females demonstrated statistically higher DMFT levels than males (p = 0.009). Negative associations were detected for education levels and untreated and extracted components (p < 0.001), and a positive association was detected for the treated caries component (p < 0.001). Permanent military personnel treated by private dentists exhibited 17.6% untreated caries, compared with 9.4% among personnel treated in the army. Officers had lower levels of untreated caries (8.6%) than others (13.3%). Among the present population, 77% had attended a dental clinic in the preceding 2 years. Permanent force personnel are offered free, comprehensive, and accessible dental treatment. The data emphasize a need for further dental health education. PMID- 10459266 TI - Lumbar epidural sympathectomy for frostbite injuries of the feet. AB - Fourteen patients with pain and paresthesias secondary to frostbite injury were treated with lumbar sympathetic blockade. The majority of patients had an excellent response to 48 hours of continuous epidural blockade with no recurrence of symptoms after blockade. Those patients who had a limited response to epidural blockade responded well to surgical lumbar sympathectomy. Lumbar sympathetic blockade is a safe and effective technique for treating the symptoms associated with frostbite injury. PMID- 10459267 TI - Improved outcome of extremely premature infants in the 1990s. AB - OBJECTIVE: To study the impact of new treatment modalities on premature infants, we compared the effects of surfactant and antenatal steroid use on outcome in two military medical centers. STUDY DESIGN: We performed a retrospective chart review of 234 infants delivered at 22 to 26 weeks estimated gestational age (EGA) at our institutions between 1986 and 1996. The patients were divided into two groups: the presurfactant group included deliveries from 1986 to 1989; and deliveries from 1990 to 1996 were in the surfactant group. RESULTS: At 23 weeks EGA or less, there was only one survivor. At 24 weeks EGA, survival was improved in the surfactant group (46% vs. 5%; p < 0.005). Survival rates at 25 and 26 weeks EGA did not differ significantly. Combined survival at 24 to 26 weeks in the surfactant group exposed to antenatal steroids was 75% versus 44% in the presurfactant group (p = 0.02) Among survivors, the incidence of grade 3 or higher intraventricular hemorrhage was less in the surfactant group (18% vs. 42%; p = 0.038). CONCLUSIONS: Use of surfactant and antenatal steroids are associated with improved survival and decreased incidence of severe intraventricular hemorrhage among the most premature infants delivered in our institutions. PMID- 10459268 TI - The medical department in military operations other than war. Part I. Planning for deployment. AB - Many military deployments are "military operations other than war" (MOOTW), a spectrum of assignments less than all-out combat. The corresponding medical support requirement differs from conventional military medical combat support and also from customary civilian medical practice. Hence, medical planners will use different doctrine and planning tools than are used in civilian facilities or on field training exercises when tasked for MOOTW activities. The deployment team must be self-sufficient, plan for very large numbers of affected individuals, and arrange for food, water, shelter, sanitation, power, light, security, transportation, communications, and team health care in advance of arrival. Careful and well-thought-out advance liaison with numerous interested parties is required to ensure mission success. The medical department on these missions may represent the lead element, and other warfare specialists and line and staff officers may support the medical mission by providing security, communications, transportation, and logistics. The medical team may find that it represents the foreign policy "point of the spear" during MOOTW deployments. PMID- 10459269 TI - Cost-effective syphilis screening in military recruit applicants. AB - A cost-effectiveness analysis of syphilis screening was performed. Strategies included no screening, universal testing at military entrance processing stations, universal testing at basic training centers, and contracting centralized screening. Probabilities derived from data retained on recruit applicants from 1989 through 1991 (N = 1,588,143) and from the published literature were used. Cost estimates were derived from costs incurred by the military and costs projected from implementing new strategies. Sensitivity analyses were performed. Modifying the existing contract for human immunodeficiency virus screening to include syphilis screening would maximize the effectiveness of screening at a cost to the Department of Defense of $9.52 per additional year of service received. The no-screening option was significantly more cost-saving than the current method of testing. Syphilis is rare and treatable, and individuals with syphilis will be identified by other means in many cases. Syphilis screening of recruit applicants at the military entrance processing stations should cease, saving the military $2,541,000 per year. PMID- 10459270 TI - Academic medical centers: a prescription for success in an era of managed care and capitation. AB - Academic medical centers (AMCs) have traditionally symbolized technological advancements and specialization in patient care. AMCs are defined as institutions that include at a minimum a hospital and associated clinics and a medical education role. Today, these institutions face a transition to managed care and radical changes in the financing of health care. These issues are not unique to the private sector health care industry. Military AMCs also must respond to many of the same changes. Their survival may rest on the ability to recognize that they can no longer ignore trends favoring lower costs, less specialization, and more primary care. This paper describes actions being taken at AMCs throughout the country to pursue innovative ways of providing accessible, high-quality, and affordable health care while maintaining quality training programs. Besides examining changes currently taking place within the academic medical industry, recommendations are offered for new initiatives. PMID- 10459271 TI - Physical therapy in a peacekeeping operation: Operation Joint Endeavor/Operation Joint Guard. AB - This article describes the deployment and utilization of physical therapy services at a combat support hospital during a recent peacekeeping mission in Bosnia. Approximately 17% of all soldiers reporting to the 21st Combat Support Hospital were evaluated and treated by physical therapy. Physical therapy services provided musculoskeletal evaluations, developed field-expedient rehabilitative programs for the deployed soldiers, and provided injury prevention programs for the peace implementation and sustainment forces. Physical therapy helped to provide a high return to duty status and a low rate of air evacuation for deployed troops. The lessons learned from this deployment can help clarify the role of physical therapy in future support operations and sustainment operations. PMID- 10459272 TI - Thoracolumbar pain among fighter pilots. AB - High +Gz forces place high stress on the spinal column, and fighter pilots flying high-performance fighter aircraft frequently] report work-related thoracic and lumbar spine pain. The aim of this study was to determine whether +Gz exposure causes work-related thoracolumbar spine pain among fighter pilots. A questionnaire was used to establish the occurrence of thoracic and lumbar spine pain during the preceding 12 months and during duties over the whole working career among 320 fighter pilots and 283 nonflying controls matched for age and sex. Thirty-two percent of the pilots and 19% of the controls had experienced pain in the thoracic spine during the preceding 12 months (odds ratio [OR] = 2.3; 95% confidence interval [CI] = 1.5-3.5; p = 0.002 for the pilots). Among the pilots, the OR increased up to 6.1 (95% CI = 1.6-23.1; p = 0.0007) with the number of +Gz flight hours. There was no difference between the groups with regard to lumbar pain during the preceding 12 months, but over their whole working careers fighter pilots (58%) had experienced lumbar pain during their duties more often than controls (48%) (OR = 1.8; 95% CI = 1.3-2.6; p = 0.002). The greater the number of +Gz flight hours, the greater the occurrence of lumbar spine pain when on duty (OR = 26.9; 95% CI = 6.2-116; p = 0.0001 for the most experienced fighter pilots). The same was not true with regard to the number of +Gz flight hours and lumbar pain during the preceding 12 months. Age had no effect on pain in the thoracic or lumbar spine. Fighter pilots flying high performance aircraft have more work-related thoracic and lumbar spine pain than controls of the same age and sex. The difference is explained by the pilots' exposure to +Gz forces. PMID- 10459273 TI - The effect of loads carried on the shoulders. AB - This work describes a way of measuring the forces acting on the shoulders when subjects carry a backpack. A frame pack was carried by 12 male subjects: 4 hikers, 4 occasional hikers, and 4 novices. They walked for 30 minutes on a 5% inclined treadmill at 3.6 km/h to simulate moderate hiking. The load was carried at T9 and was increased (10, 15, and 20 kg) after 1, 20, and 25 minutes. The data from force transducers placed on the shoulder straps were collected for 30 seconds at 100 Hz. Shoulder strain appeared to be the limiting factor in load carriage. The optimal carrying method depends on the person, but it has been shown that decreasing stride length and wearing appropriate footwear reduces the strain on the shoulders. PMID- 10459274 TI - Ultrasonography in the evaluation of hemoperitoneum in war casualties. AB - The aim of this study was to evaluate the sensitivity, specificity, accuracy, and positive and negative predictive values of emergent ultrasound examination in the detection of hemoperitoneum among war casualties, and to compare the results of this method in a specific war situation and civil conditions. Ninety-four wounded individuals with suspected blunt or penetrating abdominal trauma were treated at a level I war hospital (group W), and 242 civilians with multiple injuries with suspected blunt abdominal trauma were evaluated at the emergency center of a university hospital (group C). All examinations were performed in less than 5 minutes with a portable ultrasonographic scanner, and typical points were scanned (Morison's pouch, Douglas and perisplenic spaces, paracolic gutter). In group W, hemoperitoneum was identified correctly in 19 patients, with three false-negative and no false-positive findings, whereas group C presented 98 true-positive results, 13 false-negative results, and again no false-positive results. We observed that ultrasonography in specific war conditions showed sensitivity of 86%, specificity of 100%, accuracy as high as 97%, positive predictive value of 100%, and negative predictive value of 96%, whereas in civil conditions the corresponding values were 88%, 100%, 95%, 100%, and 91%, respectively. The sensitivity, specificity, accuracy, and positive and negative predictive values of emergent ultrasound examination in the diagnosis of hemoperitoneum are approximately equal in war and civil conditions. PMID- 10459275 TI - Digital nerve repair by autogenous vein graft in high-velocity gunshot wounds. AB - Gunshot wounds to the hands are high-energy injuries that cause widespread tissue damage, including to the nerves. Great difficulty is encountered in later reconstruction with nerve grafting of gaps in these destructive and scarred wounds. We present our experience with three patients with digital nerve repair by autogenous vein graft performed at an early stage in this type of injury. Based on our experience and that of others, this simple and rapid technique suggests a high rate of satisfactory results. It also avoids extensive and destructive late dissection and the morbidity associated with other late reconstructive procedures. PMID- 10459276 TI - Risperidone as an adjunct therapy for post-traumatic stress disorder. AB - Risperidone, an atypical antipsychotic medication, has gained wide acceptance as a first-line drug for several indications. Recently, interest has arisen in the use of risperidone as an adjunct therapy in post-traumatic stress disorder (PTSD). PTSD is a possible sequela of exposure to traumatic events. The military population, because of occupational exposure, is at a higher risk than the general population to develop PTSD, and the treatment of PTSD may be a significant issue for military practitioners. PTSD may be complicated by social issues and comorbid psychiatric conditions, and it is often difficult to treat. We present two case reports on the use of risperidone in the treatment of the intrusive thoughts and subsequent emotional reactivity experienced by some PTSD patients, along with a discussion of some possible mechanisms of action for the efficacy of risperidone in the treatment of these symptoms. PMID- 10459277 TI - May-Hegglin anomaly in a pregnancy complicated by intrauterine growth restriction and ambiguous genitalia. AB - OBJECTIVE: Thrombocytopenia as a hematologic disorder complicates up to 4% of all pregnancies. May-Hegglin anomaly is a rare cause of low platelets in pregnancy. METHODS: A case of May-Hegglin anomaly complicating pregnancy and intrauterine growth restriction in a fetus with ambiguous genitalia is described. RESULTS: The antepartum and intrapartum diagnosis and management of a patient diagnosed with May-Hegglin anomaly is discussed. The involvement and consultation of a perinatologist, neonatologist, internist, and anesthesiologist is reviewed, with emphasis on the mode of delivery. CONCLUSION: The potential maternal and fetal complications associated with May-Hegglin anomaly warrant early pregnancy diagnosis and access to a tertiary care facility. PMID- 10459278 TI - Births and deaths: preliminary data for July 1997-June 1998. AB - OBJECTIVES: This report presents preliminary data on births and deaths in the United States from the National Center for Health Statistics (NCHS) for the 12 months ending June 1998. U.S. data on births are shown by age, race, and Hispanic origin of mother. Natality data on marital status, prenatal care, cesarean delivery, and low birthweight are also presented. Mortality data presented include leading causes of death and infant mortality. METHODS: Data in this report are based on more than a 99-percent sample of births and on more than an 89-percent sample of deaths in the United States for the 12 months ending June 1998. The records are weighted to independent control counts of births, infant deaths, and deaths 1 year and over received in State vital statistics offices from July 1997 to June 1998. Unless otherwise indicated, comparisons are made with final data for the 12-month period ending June 1997. RESULTS: For the period July 1997-June 1998, the birth rate for teenagers dropped 4 percent to 51.5 births per 1,000 women aged 15-19 years, the lowest level since 1987. Birth rates for teenagers have been declining since 1991. Birth rates for women aged 20-29 years changed very little, whereas rates for women in their thirties and forties rose 2 to 4 percent. The birth rate for unmarried women declined slightly, but the number of births to unmarried women was up about 1 percent because of an increase in the number of unmarried women. The rate of prenatal care utilization continued to improve. The percent of births delivered by cesarean section rose from 20.7 percent to 20.9 percent as the result of a slight increase in the primary cesarean rate and a substantial decline in the rate of vaginal births after previous cesarean (VBAC). The overall low birthweight rate was unchanged at 7.5 percent. Age-adjusted death rates reached a record low, 2 percent below the rate for the previous 12-month period. The largest declines in estimated age adjusted death rates among the leading causes of death were for Human immunodeficiency virus (HIV) infection (37 percent) and homicide (9 percent). Smaller declines were noted for most of the other leading causes of death, but no increases occurred. Mortality also decreased for firearm injuries and alcohol induced deaths. The infant mortality rates for all races and white and black infants were about the same as the corresponding rates for the previous 12-month period. PMID- 10459279 TI - Mortality from Alzheimer's disease: an update. AB - Alzheimer's disease is a progressive degenerative condition that has devastating implications for those afflicted. An estimated 4 million Americans, mainly elderly, have this condition, which is characterized by forgetfulness in early stages and increasingly severe debilitating symptoms as the disease progresses over what can be as long as a 20-year period. As an individual's impairment increases, informal or formal care giving becomes necessary to take care of basic needs. Annually, an estimated $80 to $100 billion dollars are spent on health care expenses or lost in wages for the persons with Alzheimer's disease or their care givers. At later stages of the disease, persons with Alzheimer's disease are bedridden and vulnerable to developing other medical conditions and dying before they would if they did not have Alzheimer's disease (1). Physicians report that Alzheimer's disease caused the death of 21,397 persons in 1996 and contributed to the death of 21,703 additional persons. This information is from death certificates completed by physicians for all deaths in the United States, a fundamental source of information on what caused death for the 2.3 million deaths in the United States. The risk of dying from Alzheimer's disease has leveled off in recent years after rapid increases in the early 1980's and subsequent slower growth in the 1990's. The trend likely reflects changes in attitudes of physicians and the public about attributing Alzheimer's disease as a cause of death as well as the availability of improved diagnostic procedures; the recent leveling in mortality trends from this condition may signal that death certificate diagnoses for Alzheimer's disease are more reliable now. Alzheimer's disease is a major cause of death, which exhibits variations by age, sex, race, and geographic area. This report provides recent mortality data on Alzheimer's disease. A previous report covers historic trends (2). PMID- 10459280 TI - [Plain X-ray diagnosis of noncardiogenic edema]. AB - Noncardiogenic edema is pulmonary edema without increased precapillary blood pressure of the lung. The causes of noncardiogenic edema include decreased blood osmotic pressure, increased permeability of basement membrane, and disturbed lymphatic drainage. We describe the plain X-ray findings of noncardiogenic pulmonary edema as they correlate with the pathophysiology of pulmonary water influx from blood vessels into the extravascular space. We discuss certain selected causes of noncardiogenic edema such as ARDS and high altitude, focusing on the differential diagnosis between cardiogenic edema and noncardiogenic edema and the causes of noncardiogenic edema. PMID- 10459281 TI - [CT findings of pulmonary infections]. AB - Although many pulmonary infections are diagnosed using chest radiographs in combination with clinical findings and are treated empirically with antibiotics, there are situations in which chemotherapy is not effective and further examinations are necessary. In those cases, CT, especially high-resolution CT (HRCT), can provide additional information about detailed morphological changes of the lung that are sometimes indicative of the causative organisms of pneumonia, such as pulmonary tuberculosis. Although it is true that the same organism can present a wide spectrum of radiological findings, it is also important for differential diagnosis to understand the basic features of pathological changes of the lung and the CT findings caused by each of the various organisms. Such CT information would also be useful in differentiating infectious pneumonia from noninfectious pneumonia. This review article presents the principal CT findings of pulmonary infections and their pathological correlations with various pneumonias, including bacterial, tuberculous, fungal, and viral pneumonias. PMID- 10459282 TI - [Effects of gadobenate dimeglumine on MRI of mouse liver metastasis model constructed by orthotopic transplantation of highly metastatic murine colon carcinoma (C38-OT7): comparison with gadopentetate dimeglumine]. AB - Twelve mice with metastatic liver tumors were divided into two groups of six, with one group administered contrast medium at 0.1 mmol/kg, and the other at 0.2 mmol/kg. Contrast medium, gadobenate dimeglumine (GD) or gadopentetate dimeglumine (GP), was administered at 0.1 or 0.2 mmol/kg via the tail vein to each animal in each group. Using a Signa Horizon 1.5-Tesla MRI unit, spin-echo transverse and coronal T1-weighted sections were obtained every 15 minutes until 2 hours after administration. The numbers of liver tumors detected on films were counted before and after the administration of contrast medium, and the liver/tumor contrast-to-noise ratio (CNR) was calculated. After the completion of MRI, livers were removed, and the number of metastatic nodules on the liver surface were counted as the number of tumors. Liver/tumor CNR rose after administration in both of the GD groups. In the GP group, liver/tumor CNR remained almost constant throughout the observation period. Relative to the number of tumors detected at optical microscopy, approximately 80% and 100% of tumors were detected at MRI after the administration of GD at 0.1 and 0.2 mmol/kg, respectively. On the other hand, approximately similar numbers of tumors were detected at MRI before and after the administration of GP. These results suggest that GD administered by intravenous injection was transported promptly to the liver, increased liver/tumor CNR, and enhanced detection performance for metastatic liver tumor. PMID- 10459283 TI - [Experimental and clinical studies of fast three-dimensional MR imaging of the heart]. AB - MRI has been utilized since its inception to study the anatomy and physiology of the heart. However, the sensitivity of MRI to motion has always posed a major challenge in imaging this organ. The purpose of this study was to develop a 3D MP RAGE technique for the heart, and to apply it clinically. In the experimental study, data acquisition timing was discussed by normal volunteers. Changes in magnetization recovery time affected imaging contrast very little in the phantom study. Fourteen adults and 21 children were examined. In the adults, MP-RAGE images were rated as high in quality in the visual estimation. In the quantitative estimation, the images provided almost the same anatomical information as those of cine MRI. In the children, MP-RAGE was useful for cases of partial anomalous pulmonary venous drainage, particularly in the evaluation of abnormal pulmonary veins. The 3D MP-RAGE technique was useful in imaging the heart because it was possible to obtain continuous views in the same cardiac cycle and to reconstruct views from any direction after the examination. PMID- 10459284 TI - [Concurrent chemoradiotherapy using protracted infusion of low-dose CDDP and 5-FU and radiotherapy for esophageal cancer]. AB - PURPOSE: We evaluated the effects and safety of concurrent chemoradiotherapy for patients with esophageal cancer. MATERIALS AND METHODS: Between March 1994 and April 1998, concurrent chemoradiotherapy using protracted infusion of low-dose cisplatin (CDDP: 3-6 mg/m2/24h), 5-fluorouracil (5-FU: 200 mg/m2/24h) and radiotherapy was given to 26 patients. The median age was 70 yr, with a range from 58 to 86 yr. With regard to TNM classification (1987), six patients were stage II, five stage III, and 15 stage IV. Radiotherapy was performed by external irradiation alone in 23 patients and external irradiation plus brachytherapy in three patients. One patient underwent surgery after a dose of 40 Gy owing to the possibility of idiopathic bleeding from the stomach. RESULTS: Locally, primary effects resulted in complete response in 11 patients (42.3%) and partial response in 15 (57.7%). Acute toxicity was primarily hematologic. Leukopenia and thrombocytopenia of grade 3 or 4 occurred in eight (30.7%) and six (23.0%) of 26 patients, respectively. In patients administered CDDP at more than 5 mg/m2/day, hemotoxicity was severe because in five of the 10 patients administered 5 mg/m2 CDDP and one of the two patients administered 6 mg/m2 CDDP, thrombocytopenia of grade 3 or 4 occurred. CONCLUSION: Protracted infusion of low-dose CDDP and 5-FU with concomitant radiation therapy is effective, but from the point of acute toxicity, the optimal dose of CDDP and 5-FU needs further investigation. PMID- 10459285 TI - [Comparison of 201Tl-SPECT and MRI using Gd-DTPA for glioma]. AB - 201Tl-SPECT was performed in 25 patients with a pathological diagnosis of glioma. The lesion-to-normal (L/N) ratio of the glioblastoma group (n = 7) was found to be higher than that of the low-grade glioma group (n = 7; Mann-Whitney U-test, p < 0.0167). 201Tl accumulation in the tumor corresponded to contrast enhancement on MRI in 95% of cases. An insufficient blood-brain barrier was considered to be the primary contributor to 201Tl accumulation. In five cases, there was a discrepancy between the extent of 201Tl accumulation and the Gd-DTPA enhanced area. In these cases, the area of 201Tl accumulation was larger than the area of Gd-DTPA enhancement. This may result from damage to the blood-brain barrier that is not severe enough to be detected with Gd-DTPA or from additional factors other than change in the blood-brain barrier. 201Tl-SPECT is able to demonstrate the extent of glioma more accurately than contrast-enhanced MRI. PMID- 10459286 TI - [Composition of vascular tree using moving-table MR angiography: development and preliminary clinical experience with a semi-automated program combining stacks of MR angiographic images]. AB - Moving-table MR angiography, combining a contrast enhanced MRA approach with table movement, has recently become available. This technique allows imaging of long, longitudinal anatomical regions of vasculature. We have developed a semi automated program combining stacks of MRA images obtained from different image stations, and applied it to the reconstruction of vascular trees in twenty-five patients with peripheral arterial occlusive disease. Using this program, continuous vascular trees, extending from the abdominal aorta to lower leg (up to 124 cm), were semi-automatically reconstructed within two minutes from six different projection angles. Extensive vascular lesions were recognized three dimensionally by moving image display mode. PMID- 10459287 TI - [Migratory pneumonitis similar to bronchiolitis obliterans organizing pneumonia after conservative treatment of breast cancer: a case report]. AB - We report the case of a 63-year-old woman who developed cough and fever with migratory lung infiltrates three months after completion of right breast irradiation following conservative surgery. Lung infiltrates were initially localized in the irradiated area, but later spread to unirradiated areas in both lungs. No cause of migratory pneumonitis other than irradiation was found, and we clinically diagnosed this case as radiation-induced migratory pneumonitis similar to BOOP, without lung biopsy. Steroid therapy resulted incomplete resolution of lung infiltrates. The reported case clearly differed from typical radiation pneumonitis. We suggest that lung irradiation might trigger the development of migratory pneumonitis with a clinical pattern similar to that of BOOP. PMID- 10459288 TI - [Endoscopic sinus surgery for unilateral chronic sinusitis]. AB - Functional endoscopic sinus surgery has become an increasingly popular treatment for chronic sinusitis. This approach is aimed at re-establishment of ventilation and mucociliary clearance of the sinuses. However, some otolaryngologists believe that the Caldwell-Luc procedure should be routinely used for unilateral chronic sinusitis, because it is often associated with the maxillary sinus carcinomas. To evaluate the state of endoscopic sinus surgery for the diagnosis and treatment of unilateral chronic sinusitis, we analyzed the cases of 39 patients with unilateral chronic sinusitis who underwent endoscopic sinus procedures. These patients were unresponsive to appropriate antibiotic management for more than 6 months. Generally, endoscopic ethmoidectomy and antrostomy were performed with preservation of the middle turbinate. After the ostium was enlarged, the maxillary sinus was cleaned and carefully inspected for the presence of associated neoplasms using 30 and 70 degree endoscopes. Preoperative computed tomography (CT), postoperatve pathologic diagnosis, fiberscopic findings of the maxillary sinus, and symptomatic improvement were evaluated. Three patients had CT evidence of bone destruction of the lateral nasal wall. Pathological diagnosis demonstrated that three patients had maxillary sinus mycoses caused by Aspergillus species, one patient had inverted papilloma, and the other 35 patients had chronic sinusits. No associated malignancy was found. Eighty-one percent of the patients had almost normal endoscopic findings of the maxillary sinus by postoperative fiberscopic examination 4 to 8 months following surgery. With an average follow-up of 26 months, 88% of the patients were judged as having significantly improved in their presenting complaints of mucopurulent rhinorrhea, nasal obstruction, and facial pain. The results of this series suggest that endoscopic sinus surgery is an effective procedure for the diagnosis and treatment of unilateral chronic sinusitis. PMID- 10459289 TI - [A clinical treatise upon the nasal septal perforation]. AB - Thirty nine cases of septal perforation of various origins were presented using clinical such as backgrounds age, gender, possible causes, and size of septal perforation evaluated by diameter. The cases included 26 males and 13 females with a mean age of 35 years range: 8 to 85 years. Possible causative factors were as follows: septal surgeries (9), other nasal and paranasal surgeries (17), nasal cautery or tamponade (2), occupational (2), collagen disease (2), inflammatory (1) and idiopathic (5). Signs and symptoms related to perforations were minor, such as stuffy nose (3), running nose (3), nosebleed (5), dry sensation (4), nasal pain (1), and asymptomatic (23). Sizes of perforations by largest diameter were small (less than 10 mm) in 7, moderate (11-20 mm) in 13, and large (more than 21 mm) in 6. The size of perforations tended to be variable, but two cases with collagen disease showed large perforations 35 mm. Although iatrogenic and idiopathic cases comprised the majority of cases (80%), such a few cases of grave significases as collagen and hematological diseases showed unusually large perforations. PMID- 10459290 TI - [Clinical analysis of parotid cancer]. AB - A clinical study was performed on 42 patients with parotid cancers initially treated in our hospital from 1972 to 1997. The five-year cumulative survival rate was 69% in the whole group and 72% in the radical surgical treatment group (n = 40). The five-year survival rates according to stage were as follows: stage I, 95% (21); stage II, 75% (4); stage III, 0% (1); and stage IV, 37% (16). The factors influencing prognosis were cases of T3 and T4 (p < 0.05), stage III and IV (p < 0.01), and cervical lymph node metastasis (p < 0.01). Regarding treatment modalities, a partial parotidectomy appeared to be a curable surgical procedure for T1 cases. However, a lobectomy is recommended for T2 cases. Furthermore our study proposed that prophylactic supraomohyoid neck dissection should be necessary for mucoepidermoid carcinoma (high-grade malignancy) and undifferentiated carcinoma of T4N0 cases. PMID- 10459291 TI - [Effect of adrenaline on vagus nerve reflexes]. AB - INTRODUCTION: Dizziness or syncope may occur during treatment of nasal disease. These symptoms are considered imputable to the vagovagal reflex, which is partly involved in neurogenic syncope. In response to trigeminal nerve stimulation, the vagus nerve causes a sudden fall in heart rate and blood pressure. Elevation in blood adrenaline level during pain or tension was noted, and its effects on vagovagal reflexes were studied. MATERIALS AND METHODS: Ten cats were used in the experiment. The vagus nerve was exposed on the right side of the neck by making an incision in the trachea, and a platinum electrode was attached to the vagus nerve on nerve's distal side. The head was tilted a 30 degree angle. The cats were divided into control and adrenaline-treated groups, and changes in cerebral blood flow, heart rate, and arterial pressure were compared between the two groups after electric stimulation of the vagus nerve. Cerebral blood flow was measured by the hydrogen clearance method. 1) Control group The vagus nerve was electrically stimulated for 1 minute. 2) Adrenaline-treated group The vagus nerve was electrically stimulated for 1 minute following 30 seconds of intravenous administration of adrenaline. RESULTS: Cerebral blood flow was significantly decreased in both the control and adrenaline-treated groups after electric stimulation, but the decrease was significantly greater in the latter group at all sites of measurement. Whereas heart rate and arterial pressure were significantly decreased in the control group, these variables in the adrenaline treated group showed no significant change despite the greater decrease in cerebral blood flow. DISCUSSION: Heart rate, blood pressure, and cerebral blood flow were all significantly decreased after electric stimulation of the vagus nerve. These changes were considered owing to a fall in blood pressure due to vasodilation resulting from bradycardia and a relative decrease in sympathetic nervous tension resulting from electric stimulation. On the other hand, in the adrenaline-treated group, neither heart rate nor blood pressure showed any significant change, but cerebral blood flow was significantly decreased at all sites of measurement despite an adrenaline load. This contradictory results may be accounted for by the powerful beta 2-activity of adrenaline. The greater decrease in cerebral blood flow in the adrenaline-treated than in the control group can be attributed to decreased peripheral vascular resistance by its beta 2 activity. In the field of otorhinolaryngology the trigeminal region is often involved in the treatment of nasal disorders so that vagovagal reflexes are often encountered. The results of this study counsels caution in the treatment involving the trigeminal region. PMID- 10459292 TI - [Effects of topical alpha 1- and beta 2-adrenoceptor stimulants on nasal nitric oxide level]. AB - The effects of locally administered alpha 1- and beta 2-stimulants (naphazoline and salbutamol) on the nasal nitric oxide (NO) level were investigated. Twenty four healthy volunteers (except nasal allergy) were subjected to the examination. First, nasal cavity air was sampled continuously from the right nostril for 20 seconds at the rate of 3.5 l/m, and NO-free air was supplied passively to the left nostril. During the sampling time, subjects were made to hold their breath at deep inspiration, which obviated the effect of lower airways by closing their glottises. The sampled air was analyzed using a chemiluminescence technique for NO detection. In addition, nasal airway resistances (NAR) were estimated by a rhinomanometer, and minimum cross-section area (MCA) and nasal cavity volume (NCV) were estimated by an acoustic rhinometer. After these estimations, 12 subjects received naphazoline nitrate 15 micrograms per nostril, and the other 12 subjects received salbutamol sulfate 100 micrograms per nostril. Finally, after 15 minutes rest, these four parameters were reviewed. The results demonstrated that naphazoline significantly decreased NO concentration and NAR, and increased NCV. Furthermore, salbutamol significantly increased NO concentration and NAR, and decreased MCA and NCV. The changes in NAR, MCA and NCV indicated that nasal mucosa became contracted and swollen by topical naphazoline and salbutamol application. Naphazoline, a nasal decongestant, contract nasal vessels by stimulating alpha 1-adrenoceptors, whereas salbutamol dilates then by stimulating beta 2-adrenoceptors, and this vasodilation does not intervene NO and cyclic GMP. Thus, nasal NO concentration is significantly affected by the change of blood supply caused simply by vasocontraction and vasodilation. In conclusion, it appeared that nasal NO concentration was possibly altered by the change of nasal blood supply, moreover, by the change in the supply of NO substrate. PMID- 10459293 TI - [Experimental studies of the protective effect of ginkgo biloba extract (GBE) on cisplatin-induced toxicity in rats]. AB - Cisplatin (CDDP) is an effective antineoplastic agent in the treatment of solid malignant tumors. Its clinical use, however, is limited because of various side effects including sensorineural hearing loss. Several agents have been proposed to reduce these side effects. GBE has recently been reported to scavenge superoxide and hydroxyl radicals, resulting in a reduction of lipid peroxidation. GBE is expected to protect against CDDP-induced toxicity because its generative mechanism is thought to be associated with free-radical formation. The purpose of the present study was to evaluate GBE's efficacy as a protective agent against cisplatin-induced ototoxicity. Fisher rats were used in this study and were divided into three treatment groups: 1) animals administered 1.0 mg of CDDP per kg for 10 days (Group I), 2) animals receiving 100 mg of GBE per kg 90 min before administration 1.0 mg of CDDP per kg (Group II) and 3) a vehicle control (Group III). First, the protective effect of GBE on CDDP-induced ototoxicity was investigated. The auditory threshold was evaluated by means of the compound action potential (CAP) recording. After CAP recordings, cochlear sensory epithelia were observed throughout the cochlea by scanning electron microscopy. In Group II, the elevation of CAP thresholds at 12 kHz, 16 kHz, 20 kHz and the missing rate for the outer hair cells were significantly reduced as compared to those in Group I. These data suggest that GBE is effective for otoprotection against CDDP. Second, the protective effect of GBE on CDDP-induced nephrotoxicity was evaluated. Nephrotoxicity was evaluated by means of measurement of serum BUN and creatinine and histopathological examination of the kidney. These were significant differences in serum BUN and creatinine levels between Group I and Group II. Third, the influence of GBE against the antitumor effect of CDDP was researched in the rats inoculated subcutaneously with SCC-158 squamous cell carcinoma cells. There was no difference in tumor growth rate (TGR) between Group I and Group II. The result suggested that the combined administration had no influence on the antitumor activity of CDDP. In conclusion, the co-administration of CDDP with GBE is beneficial to ameliorate CDDP-induced toxicity without attenuation of CDDP antitumor activity. PMID- 10459294 TI - [A prospective study of introducing self-determined treatment policy for the patients with hypopharyngeal cancer]. AB - A total of 57 patients with hypopharyngeal cancer were given an opportunity to select their treatment options after a thorough explanation of the tumor stage, treatment modalities, complications due to treatment, and prognosis. As a result, many patients selected chemoradiotherapy as the initial treatment. Two patients requested treatment at other institutes and were immediately transferred. A 91 year-old patient with T4N3M0 tumor declined treatment. The duration from the initial visit to the start of treatment course was prolonged because of the informed consent process. Patient rejection of salvage surgery following the failure of chemoradiotherapy increased. However, both the physician-patient relationship and physician-patient's family relationship in the uncured group did not deteriorate. No change in the 3-year crude survival rate by introducing self determined treatment was observed at this time. It is ideal that the patient choice of treatment options is the one physician recommend. Meanwhile, physicians should consider ideal treatment options, which are most desirable for the patient. In conclusion, thorough informed consent provides positive clinical impact even with ominous hypopharyngeal cancer. PMID- 10459295 TI - [The efficiency of magnetic resonance angiography (MRA) in the diagnosis and vertigo--prediction of vertebrobasilar insufficiency (VBI) and atherosclerosis]. AB - Major causes of vertigo in patients who attend Otolaryngology clinics are peripheral vestibular disorders (PVD) and vertebrobasilar insufficiency (VBI). The purpose of this study was to see whether MRA findings can distinguish VBI from PVD to evaluate the efficiency of MRA in the diagnosis of vertigo. A total of 185 vertigo patients were examined by MRA with the 3D-Phase Contrast method. Three kinds of abnormalities of vessels, that is, (a) disappearance, (b) meandering, and (c) stoppage were found in MRA imaging. Therefore, we classified the MRA patterns into four types with two subtypes: type I; normal, type II-R; right hypoform of the vertebral artery, II-L; left hypoform of the vertebral artery, III; meandering form of the vertebrobasilar artery, IV-1; stoppage form of the basilar artery, IV-2; hypoform of the bilateral vertebral and basilar arteries. Diagnosis of the MRA patterns in each patient was camed out by a radiologist and neurosurgeon with the cooperation of an otolaryngologist. In 185 vertigo patients, 139 patients were clinically diagnosed as having PVD, 41 patients as having VBI, and 5 patients as having cerebellar and brainstem infarctions. The numbers of patients in MRA patterns I, II-R, II-L, III and IV were 140, 17, 12, 8, and 8 cases, respectively. The total number of VBI patients who demonstrated type III or IV patterns in MRA was significantly higher than that of type I, II-R and II-L (P < 0.005). All of the 8 cases with cerebellar and brainstem infarction belonged to type IV. Cerebral angiography was performed in nine cases with type II-R, II-L, III or IV-1, and the number of patients in each group was 3, 3, 1 and 2 cases, respectively. All of the type II-R and II-L cases revealed hypoform of the vertebral artery. The appearance of type III was restricted to type III's view was only meandering and type IV-1 showed severe stenotic changes in the union area. These data show that MRA findings in the diagnosis of vertigo patients are very effective in distinguishing VBI and cerebellar and brainstem infarction from PVD and that some pattern of MRA may correspond to atherosclerosis of the vertebrobasilar artery. We think MRA examination of vertigo patients is useful not only to assist in the diagnosis of the etiology but also to predict the stage of atherosclerosis in each patient. PMID- 10459296 TI - [Segment transport]. PMID- 10459297 TI - [Tibial segment transport]. PMID- 10459298 TI - [Injuries due to falls from a great height. A comparative analysis of injuries and their outcome following suicide-related and accidental falls]. AB - The aim of this study was to compare the outcome and clinical course of multiple trauma patients with accidental or intentional (suicide related) fall from heights > 4 m. 211 patients with an injury severity score (ISS) > 17 were assigned to the following groups: I: intentional fall, n = 94; A: accidental fall, n = 117) and ISS (I: 28 +/- 1; A: 30 +/- 1), ventilation time (I: 16 +/- 2; A: 15 +/- 1) were not different. Significant differences were found in sex (m/f: I: 56/44; A: 73/27%), fractures of lumbarspine (I: 34; A: 15%), pelvis (I: 51; A: 38%), lower leg (I: 47; A: 20%), pilon (I: 15; A: 5%), and os calcis (I: 17; A: 9%). Liver lacerations occurred more often after intentional fall (I: 16; A: 6%). Single or multiple organ failure (MOF) was diagnosed significantly more often in group A(I: 1; A: 8%). Main cause of death in both groups was single or multiple organ failure (MOF: I: 47; A: 69%) or related to brain-injuries (I: 35; A: 19%). Prognosis and rehabilitation of multiple trauma patients after intentional fall is related to brain-injuries, spine-fractures and the functional outcome of the injured lower leg. Prognosis of patients after accidental fall is related to the development of MOF during the ICU-course. PMID- 10459299 TI - [Integration of porous hydroxyapatite ceramic prosthesis at the distal radius in elderly patients. Radiological examination]. AB - Between April 1994 and December 1995 14 patients (average age 56 (41-71) years, 13 women, one man) suffering from a fracture of the distal radius (Extensionsfracture, type 1B by Pechlaner, AO-classification A2 and A3) were treated by minimal invasiv procedure. The bone defect was substituted with hydroxylapatit ceramic. In a prospective study the osteointegration of the alloplastic bone graft was evaluated radiographically and on MRI. Up to week 12 X ray revealed a radiolucenty around the allograft in all patients. After 6 months trabecular structure reached the alloplastic bone graft in 57% of the patients. After one year the surface of the hydroxylapatit ceramic was remodelled in 36% of the patients. At this time MRI examination with contrast agent demonstrated marginal integration of the alloplastic bone craft in 71%. X-ray follow up showed stageable integration of the porous hydroxylapatit ceramic. MRI examination demonstrated partial osteointegration. Hydroxylapatit ceramic was incorporated in radial metaphyse of elder patients without any complications. PMID- 10459300 TI - [The external patello-tibial transfixation (EPTT). Part II: Clinical application and results]. AB - In part I of the paper the biomechanical and technical background of the EPTT using the MPT fixator and the indications for this procedure have been described. In part II we report about the clinical application of the EPTT in 67 patients with a wide spectrum of repairs and reconstructions of the extensor mechanism. 48 patients had fresh injuries, 18 of them with severe concomitant knee lesions and 19 patients had neglected rsp. unsuccessfully operated injuries. There were 4 deep infections, two of them related to the MPT fixator. In the patients with uneventful healing the fixator remained in place for 7.3 weeks in average. The clinical, isokinetic and radiological results were reviewed in 17 patients with an average follow-up time of 37.3 months. There were 5 patients with partial patellectomy and tendon reattachment because of lower patella pole comminution and 12 patients with tendon reattachment ruptured at the inferior patella pole or suture repair in midsubstance rupture. The clinical results according to the IKDC score were rated in 3 patients as normal, in 10 patients as nearly normal and in 4 patients as abnormal. This rating was highly dependend on the subjective judgement by the patients who considered their operated knees not as normal as the contralateral knees. From our clinical experiences and results we can derive that the EPTT enables the surgical management of extensor mechanism disruptions with a minimum of internal fixation material and provides a safe protection of the repairs and reconstructions during the healing period. The EPTT allows immediate unrestricted functional rehabilitation and early walking without crutches. Thus the EPTT represents an effective alternative to the patello-tibial cerclage with a wire or synthetic ligaments. PMID- 10459301 TI - [Blood supply of the quadriceps tendon]. AB - Degenerative changes have been considered to be a cause for spontaneous quadriceps tendon rupture. Aim of this study is to investigate the microvasculature of the quadriceps tendon by injection techniques and immunohistochemical methods (antibodies against laminin) with regard to the pathogenesis of tendon degeneration. The blood supply of the quadriceps tendon arises from descending branches of the lateral circumflex femoral artery, by branches of the descending geniculate artery and by branches of the medial and lateral superior geniculate arteries. Blood vessels penetrate the tendon from the surrounding connective tissue and anastomose with a longitudinally orientated intraligamentous network. Compared to the surrounding synovial layer, the amount of vessels in the tendon substance is greatly reduced. The distribution of blood vessels within the quadriceps tendon is not homogenous. The anterior or superficial part of the tendon has a complete vascular network that extends from the musculo-tendinous junction to the patella. Within the deep portion of the quadriceps tendon there is an oval shaped avascular area which measures app. 30 mm in length and app. 15 mm in width. Within this area the immunohistochemical proof of laminin is negative. An explanation for the absence of blood vessels may be compressive stress caused by the patellar groove which serves as hypomochlion for the quadriceps tendon when the knee is flexed. The occurrence of an avascular zone within the deep layer of the tendon that is directed to the joint cavity may explain the frequency of degenerative changes in this region. PMID- 10459302 TI - [Reliability of functional x-ray analysis of cervical vertebrae flexion and extension]. AB - Functional radiographic analysis of the cervical spine in flexion and extension position is increasingly used as screening method for the diagnosis of segmental functional disorders. The objective of this study was to prove Penning's evaluation method for the metrical recording of segmental angles in the sagittal plane for selectivity, reliability and usability. Passive functional radiographs of the cervical spine in flexion and extension were taken of 20 patients with painful limitation of mobility of the cervical spine and 20 subjects, similar in sex and age, without complaints. The radiographs were duplicated. Five physicians measured the angles of segmental mobility in a blind study. Statistical analysis was conducted using the t-test and calculating the correlation coefficient "r". The results of the study prove that the evaluation method by Penning shows a usable and, for segments C3/C4 to C6/C7, significant selectivity. The selectivity of p < or = 0.05 and p < or = 0.01 is sufficient to distinguish patients from healthy subjects. The correlation between the five reviewers showed good to very good results (0.6 < r < or = 0.8; r > 0.8). The measured values, however, have to be considered, in connection with the appropriate clinical symptoms, as still "normal" or "functionally disordered" in the context of segmental hypo-resp. hypermobility. PMID- 10459303 TI - [The unstable elbow joint]. PMID- 10459304 TI - [Ischemia-related small intestine perforation 15 days after car accident. Fate or avoidable complication?]. AB - Intraabdominal organ lesions after blunt abdominal injuries often are missed, especially in the contest of a polytrauma because of the absence of obvious clinical injury signs, difficult examination conditions of an intubated patient and the possibility of secondary development of lesions. For an exact recognition of the abdominal injuries a standard diagnostic procedure is necessary. Clinical and laboratory parameters which determine the circulatory situation, initial abdominal ultrasound, x-ray and follow up examinations are indispensable. In equivocal situations CAT scan respectively angiography are recommended. The correct initial diagnosis of intestinal perforations still is problematic and requires repeated examinations. We report the case of a patient with initial blunt abdominal injury and a non dislocated Weber-A-fracture after a car accident. 15 days after trauma the patient was seen with the clinical signs of an acute abdomen. After emergency laparotomy an ischemic necrosis of the ileum, covered with net, was seen. PMID- 10459305 TI - [Solitary juvenile cyst of the heel bone. Two case reports and review of the literature]. AB - The calcaneus is a uncommon site for a unicameral solitary bone cyst. However, because of the concentration of forces through the heel, such solitary bone cysts are usually symptomatic, depending on the proximity to the joint and the size of the cyst and therefore require in most cases surgical treatment. This is a report about two patients with a solitary bone cyst of the os calcis in 1996 and 1997. The diagnosis was confirmed radiologically and histologically. The standard therapie in both cases was the curretage of the cyst by packing the defect with bone graft from the pelvis. The author reviewed the literature concerning ethiology, diagnosis and treatment of unicameral solitary bone cysts of the calcaneus. The diagnosis cannot be based solely on radiological findings because of the variation of solitary bone cysts and the special forms, such as calcifying solitary bone cysts. Therefore exact histological diagnosis is of particular importance. PMID- 10459306 TI - [Intraoperative extracorporeal irradiation and replantation (EIR) in the treatment of primary malignant bone tumors]. AB - In 6 patients with primary malignant bone tumors (4 Ewing, 1 recurrent Adamantinoma, 1 maligne Haemangioperizytoma) the local therapy was performed as intraoperative extracorporeal irradiation and replantation. During the follow-up 10 (6-24) months no local recurrence in the replanted segment occurred. One recurrence occurred in the host bed; in two cases the replanted segment had to be removed because of severe infection. The other cases showed good functional and radiological results. These clinical findings and the reports in the literature seem to allow the application of the intraoperative extracorporeal irradiation and replantation procedure for defect reconstruction in cases of chemo- and radiotherapy-sensitive bone tumors. PMID- 10459307 TI - A database designed for utilization management in diagnostic imaging. AB - The methods and tools of health services research have been applied to a diverse number of health care areas. Surprisingly, they have been adopted only recently in diagnostic imaging, by a small number of professionals, in response to the severe fiscal constraints and widespread structural changes in the health industry, as well as to a growing concern that the value of social and individual investment in high-cost imaging services could not be validated objectively. As a result of the need for accountability for the use of scarce resources, regulators and payers of health services increasingly demand that a reasoned and objective evaluative process be adopted. To undertake a statistically driven evaluative approach that stands up to objective assessment of methodological rigour, an organized data-collection system is needed. Without this fundamental cornerstone, evaluators are left with little more than anecdotal evidence and professional and personal opinion to guide decision-making. It then becomes difficult to learn from both the successes and failures that are routinely experienced during times of rapid and fundamental change. This article describes the efforts made to integrate health services research in radiology into the routine daily activities and supporting systems of a large academic health system, the Hamilton Health Sciences Corporation and McMaster University Department of Radiology, in an attempt to move in the direction of evidence-based decision-making. The authors hope this will allow others to learn and improve on this work. Radiologists may then move the vast data systems and infrastructure associated with all imaging services to an evidence-based model for managing and guiding the vast resources entrusted to our collective stewardship. PMID- 10459308 TI - The radiologist and the patient: breaking bad news. AB - Radiologists may be asked to disclose results directly to patients. Studies of radiologist-patient communication show that radiologists have a direct responsibility to their patients, that many patients wish to learn the results of imaging tests from the radiologist at the time of the examination, and that many referring physicians support the principle of radiologists disclosing results when patients ask for them. In some areas, such as breast imaging and interventional radiology, disclosure by radiologists is very common. The authors, who work in a perinatology unit in which obstetrical ultrasonography is performed, have developed a template to help them with the often-emotional interactions associated with pregnancy failure. They recommend that radiologists inform the patient clearly of the examination results, choosing everyday words such as "miscarriage" and "pregnancy." They also recommend that physicians show compassion, acknowledge patients' grief and inform the referring physician as soon as possible. These steps should be part of a policy of direct, honest communication with patients. PMID- 10459310 TI - Sonographic surveillance of abdominal aortic aneurysms: what is the smallest change in measured diameter that reliably reflects aneurysm growth? AB - OBJECTIVE: The decision to repair an abdominal aortic aneurysm may be based on an apparent increase in aneurysm diameter seen in successive sonographic examinations. However, true aneurysm growth can be diagnosed only if the measured increase in aneurysm diameter exceeds the variability inherent in the measurement. This study uses analysis of variance to determine the smallest change between 2 successive, independent sonographic measurements of aneurysm diameter that reliably indicates aneurysm growth. METHODS: Pairs of independent observers examined 63 patients with abdominal aortic aneurysms. Each observer obtained a sonographic measurement of the anteroposterior aneurysm diameter in the transaxial and longitudinal scan planes, and the transverse aneurysm diameter in the transaxial scan plane. Analysis of variance yielded the total variance associated with each type of measurement as well as the contributions made by variance among aneurysms, variance between observers and residual variance. These components were used to estimate interobserver standard error of measurement, interobserver reliability and the smallest measurement change needed to diagnose true aneurysm growth. RESULTS: Differences among aneurysms made the largest contribution to overall variance. Interobserver reliability was excellent, ranging from 0.89 to 0.94 (with perfect reliability being 1.00). The smallest difference between 2 successive, independent anteroposterior diameter measurements that indicated aneurysm growth at the 95% confidence level was 0.78 cm in the transaxial scan plane and 0.92 cm in the longitudinal scan plane. The smallest difference between 2 successive, independent transverse diameter measurements that indicated aneurysm growth at the same confidence level was 1.05 cm. CONCLUSION: Despite high interobserver reliability, only changes in measured aneurysm diameter greater than or equal to 0.78 cm indicate aneurysm growth at the 95% confidence level. Smaller changes in measured diameter may not be real, but due to variability in measurement. PMID- 10459309 TI - Mammographic pattern of microcalcifications in the preoperative diagnosis of comedo ductal carcinoma in situ: histopathologic correlation. AB - OBJECTIVE: The comedo subtype of ductal carcinoma in situ (DCIS) is more aggressive than noncomedo DCIS. Differentiating noncomedo DCIS from the more aggressive comedo subtypes on mammography would allow the surgeon to excise comedo DCIS with a wider margin. The mammographic features of microcalcifications associated with nonpalpable comedo DCIS, noncomedo DCIS and benign disease were compared to determine the usefulness of this finding in diagnosis of comedo DCIS. METHODS: The authors retrospectively and blindly reviewed the mammograms of 91 consecutive patients in whom DCIS was diagnosed by needle localization and surgical excision. An equal number of cases of benign microcalcifications were also reviewed. Microcalcifications were evaluated with respect to pattern, density, configuration and size. These results were correlated with the pathologic findings. RESULTS: All 16 cases (100%) of linear branching calcifications and 34 (80%) of the 43 cases of linear calcifications were associated with comedo DCIS (p < 0.001). The number of calcifications, the density and the size of clustering were not diagnostic of comedo DCIS. Granular calcifications occurred in noncomedo DCIS and in benign disease associated with noncalcifying DCIS. CONCLUSION: Comedo DCIS is suggested by the presence of linear and linear branching microcalcifications on mammography. PMID- 10459311 TI - Extensive transmural myocardial calcification: case report. PMID- 10459312 TI - Acute afferent loop obstruction diagnosed with computed tomography: case report. PMID- 10459313 TI - Transabdominal power Doppler sonography of the normal early placenta. AB - OBJECTIVE: To evaluate how well transabdominal power Doppler sonography, a new method of colour Doppler sonography, visualizes the vasculature of the early placenta. METHODS: Evaluation of intrachorionic/intraplacental vasculature in 15 normal, singleton gestations of 8 to 18 weeks. RESULTS: Intrachorionic/intraplacental vessels were identified in all gestations, except the earliest (8 weeks). The number of vessels detected increased with increasing gestational age. Intrachorionic/intraplacental Doppler waveforms of maternal arterial origin were identified in gestations of 8.5, 9.5, and 11.5 weeks. CONCLUSIONS: Transabdominal power Doppler sonography detects intrachorionic/intraplacental vasculature at least as early as 8.5 weeks. Intervillous arterial flow of maternal origin is present in the first trimester, confirming the results of a recent transvaginal colour Doppler sonographic study, and contradicting earlier studies suggesting that such flow is not present in the first trimester. PMID- 10459314 TI - Percutaneous gastrostomy for enteral nutrition: long-term follow-up of 176 procedures. AB - OBJECTIVE: To evaluate the efficacy and safety of percutaneous gastrostomy (PG), using small catheters without gastropexy, to deliver enteral nutrition. METHODS: We reviewed the records of 176 consecutive patients in whom PG was attempted for enteral nutrition. Of these cases, 172 catheters were inserted by the Seldinger technique under fluoroscopic guidance alone, 2 were inserted under computed tomographic guidance, and 2 procedures failed. In primary procedures, 8.5- or 10.2-French catheters were used. RESULTS: The technical success rate was 98.9%. Of the 176 procedures, 2 failed because of the high position of the stomach. Seven-day follow-up was obtained in all patients; 30-day follow-up in 145 patients (83%), and long-term follow-up (30 to 1512 days) in 123 patients (70%). The 30-day mortality rate was 13.2%. One death (0.5%) was directly related to the procedure. Major complications occurred in 4 patients (2.2%), and minor complications in 12 patients (6.9%). No patient required surgery for complications attributable to the gastrostomy procedure. CONCLUSION: PG without gastropexy using small catheters is a simple, safe and effective technique for delivering enteral nutrition. PMID- 10459315 TI - Therapeutic embolization of a pseudoaneurysm of the superior gluteal artery occurring as a complication of bone marrow biopsy: case report. PMID- 10459316 TI - Safety of outpatient arterial stenting. AB - OBJECTIVE: To assess the safety of performing iliac arterial stenting as an outpatient procedure. METHODS: Retrospective analysis of safety including all patients referred for elective iliac arterial stenting over a 1-year period. Sources of data for the analysis included pre- and post-stenting vascular surgical consultation records, hospital case notes, diagnostic and interventional angiography reports, computerized laboratory data, nursing records from our angiography holding area, and the results of routine post-stenting telephone follow-up. RESULTS: There were 29 outpatient iliac stenting procedures in 28 patients (19 men and 9 women, age range 41.0 to 79.8 years, mean age 66.1 years). Of these 29 procedures, 17 involved unilateral iliac stenting, and 12 involved bilateral iliac stenting. Adjunctive renal artery angioplasty was performed in 1 patient and internal iliac angioplasty and stenting were performed in 2 patients. A total of 51 stents were deployed through 42 femoral punctures via introducer sheaths ranging in size from 6 to 8 French. Percutaneous hemostatic closing devices were used in 6 punctures. Two patients required overnight inpatient observation for moderate-size hematomas; these had no clinical sequelae. All others were discharged safely 5 to 6 hours after sheath removal. No clinically significant sequelae were identified in any patient. CONCLUSION: Arterial stenting can be performed safely on an outpatient basis. PMID- 10459317 TI - Residents' corner. Answer to case of the month #62. Systemic mastocytosis. PMID- 10459318 TI - Residents' corner. Answer to case of the month #63. Inflammatory aneurysm of the abdominal aorta. PMID- 10459319 TI - Indications for surgery in our global village. PMID- 10459320 TI - Soft-tissue images. Intestinal nonrotation in the adult. PMID- 10459321 TI - Musculoskeletal images. Granuloma of the buttock after uncemented total hip arthroplasty. PMID- 10459322 TI - Soft-tissue case 28. Ectopic thyroid. PMID- 10459323 TI - Musculoskeletal case 5. Hemangioma of bone. PMID- 10459324 TI - Ovarian carcinoma of low malignant potential treated at the Jewish General Hospital, Montreal, between 1973 and 1997. AB - OBJECTIVE: To review the epidemiologic and pathological characteristics and the management of ovarian cancer of low malignant potential (LMP) at a university teaching institution. DATA SOURCE: Hospital charts from 1973 to 1997. DATA EXTRACTION: The authors carried out a manual study of the individual hospital charts covering the study period. DATA SYNTHESIS: The findings of this review revealed that the mean age of the 30 women in the study was 48.7 years and was similar in the subgroups of women having serous (18) and mucinous (9) types. In those women for whom staging information was available, all had either stage I disease (12 serous, 7 mucinous) or stage III disease (4 serous, 1 mucinous). Treatment consisted of: total abdominal hysterectomy (TAH) and bilateral salpingo oophorectomy (BSO) with or without omentectomy (OM); BSO, unilateral oophorectomy or ovarian cystectomy alone; or TAH, OM and left salpingo-oophorectomy in women with stage I tumours. All women with stage III tumours underwent TAH, BSO and OM. The recurrence rate was low. Only 1 of 22 stage I tumours but 3 of 5 stage III tumours recurred. CONCLUSIONS: Appropriate postoperative treatment for women with this type of ovarian cancer should be conservative. However, the management of higher stage disease remains controversial. PMID- 10459325 TI - Molecular alterations in bone and soft-tissue sarcoma. AB - Understanding the molecular alterations evident in human cancer offers the potential for more meaningful diagnosis and more accurate prediction of cancer risk as well as the hope of developing novel therapies. In this review, several of the molecular abnormalities identified in both bone and soft-tissue sarcoma are outlined. Some sarcomas exhibit consistent gene translocations, which produce a unique oncoprotein that can serve as a molecular fingerprint for the sarcoma subtype. Mutations in tumour suppressor genes result in cell proliferation that is not regulated by normal cell cycle control genes. Amplification (multiple copies) of genes that act as positive controls on cell division have a similar effect on increasing cell proliferation. Finally, the understanding of drug resistance and sarcoma response to signalling systems such as IGF ligand and receptors may help in designing new therapies based on molecular changes in the cancer. PMID- 10459326 TI - Presidential address 1998. In search of daylight. AB - Practising medicine in Canada has become increasingly bureaucratic, confrontational and stressful. The Canadian Orthopaedic Association must take a far more proactive role in the development of orthopedic surgeons as professionals and in the political environment in which they practise. Living in a "knowledge-rich workplace" orthopedic surgeons must support continuous professional development and provide leadership and incentive to maintain competence in their profession. The "baby boomers" are coming. Their numbers will have a profound effect on the practice of orthopedic surgery, not 20 or 30 years from now but within the next 10 years. Therefore it is imperative that orthopedic surgeons assess and accept the impact that the "boomers" will have on surgeons, hospital beds and operating-room time. Orthopedic surgeons and the Canadian Orthopaedic Association are challenged by a new role as vendors of information in a new "information age" economy, whose fundamental sources of wealth are knowledge and communication. PMID- 10459327 TI - Treatment of osteonecrosis of the femoral head by free vascularized fibular grafting: an analysis of surgical outcome and patient health status. AB - OBJECTIVE: To evaluate the limb-specific outcome and general health status of patients with osteonecrosis of the femoral head treated with vascularized fibular grafting. DESIGN: A retrospective review. SETTING: A single tertiary care centre. PATIENTS: Fifty-five consecutive patients with osteonecrosis of the femoral head who underwent fibular grafting (8 bilaterally). INTERVENTION: Vascularized fibular grafting. OUTCOME MEASURES: Limb-specific scores (Harris Hip Score, St. Michael's Hospital Hip Score), general health status (Nottingham Health Profile, SF-36 health status survey) and radiographic outcome measures (Steinberg stage). RESULTS: Patients were young (mean age 34 years, range from 18 to 52 years) and 80% had advanced osteonecrosis (Steinberg stages IV and V). Fifty-nine hips were followed up for an average of 50 months (range from 24 to 117 months) after vascularized fibular grafting. Sixteen hips (27%) were converted to total hip arthroplasty (THA). To date, 73% of hips treated with vascularized fibular grafting have required no further surgery. Preoperative and postoperative Harris Hip Scores were 57.3 and 83.6 respectively (p < 0.001). As measured by patient oriented health status questionnaires (SF-36, Nottingham Health Profile) and compared with population controls, patients had normal mental health scores and only slight decreases in physical component scores. CONCLUSIONS: Free vascularized fibular grafting for osteonecrosis of the femoral head provides satisfactory pain relief, functional improvement and general health status and halts the progression of symptomatic disease. PMID- 10459328 TI - Randomized, controlled trial of bupivacaine injection to decrease pain after laparoscopic cholecystectomy. AB - OBJECTIVES: To determine if intraoperative instillation of bupivacaine would decrease early postoperative pain after laparoscopic cholecystectomy, if the patients would consequently require less narcotic postoperatively and if such patients would elect to be discharged on the day of operation if given the choice. DESIGN: Double-blind, randomized, controlled trial. SETTING: A tertiary care hospital in Hamilton, Ont. PATIENTS: Fifty patients underwent laparoscopic cholecystectomy. Day-surgery patients had the choice of staying overnight for discharge the following day. They were compared with a control group of 47 patients who had laparoscopic cholecystectomy but did not receive bupivacaine. INTERVENTION: Instillation of 20 mL of 0.5% bupivacaine with epinephrine into laparoscopic cholecystectomy port sites intraoperatively before closure. MAIN OUTCOME MEASURES: Visual analogue scale (VAS) pain scores assessed 4 times postoperatively, the choice of patients to leave hospital the same day or to remain in the hospital overnight; the level of postoperative narcotic usage. MAIN RESULTS: Mean VAS pain scores (range 0 [no pain] to 5 [severe pain]) at less than 2 hours and at 6 hours after surgery were 2.9 and 2.9, respectively, in the bupivacaine group compared with 4.5 and 4.0, respectively, in the control group (p = 0.001 and 0.025). VAS scores at 10 hours postoperatively and the next morning did not differ between the groups. More patients in the bupivacaine group elected to go home on the day of surgery (p = 0.034). Narcotic usage was not significantly different. CONCLUSION: Instillation of bupivacaine into port sites should be standard practice for elective laparoscopic cholecystectomy. PMID- 10459329 TI - Sickle cell disease of the spine in children. AB - OBJECTIVE: To determine the incidence of back pain in children admitted with sickle cell disease so as to increase awareness of this disease in black children presenting with back pain or discomfort. DESIGN: A retrospective review. SETTING: The Children's Hospital of Eastern Ontario in Ottawa, a tertiary care, university affiliated centre. PATIENTS: Thirty children were treated for active sickle cell disease between 1990 and 1996. Eleven (5 boys, 6 girls) suffered vascular occlusive phenomena occurring in bone, referred to as "bony crises," requiring a total of 49 admissions. MAIN OUTCOME MEASURES: Clinical manifestations of spinal involvement by sickle cell disease. MAIN RESULTS: The spine represented the second most common area of bone involvement (26%) exceeded only by the knee (35%). The vertebral level affected was lumbosacral in 66% of cases, followed by thoracic in 22% and cervical in only 12%. Eighty-six percent of the children with spinal pain were anemic upon presentation, 71% had an elevated leukocyte count, 15% were hyponatremic and 15% were hyperkalemic. Minimal physical signs in the spine were noted, other than a local tenderness over the spinous process in 71% and a decreased range of back motion in 17%. CONCLUSIONS: Sickle cell disease is becoming more common in Canada as a result of increasing immigration from African countries and should always be considered as a possible cause of back pain in a black child. PMID- 10459330 TI - Evaluation of laparoscopic skills: a 2-year follow-up during residency training. AB - OBJECTIVE: To evaluate laparoscopic technical skill in surgical residents over a 2-year period. DESIGN: The laparoscopic technical skills of general surgical residents were evaluated using the MISTELS program. This provides an objective evaluation of laparoscopic skill, taking into account precision and speed. SETTING: Inanimate laparoscopic skills centre. PARTICIPANTS: Ten general surgical residents (5 PGY1, 3 PGY2 and 2 PGY3 residents) who were required to complete 3 structured laparoscopic tasks. OUTCOME MEASURES: A composite score incorporating precision and timing was assigned to each task. The paired t-test was used to compare performance of each resident at the 2 levels of their residency training for each task. Linear regression analysis was used to correlated level of training and total score (sum of all tasks). RESULTS: Linear regression analysis demonstrated a highly significant correlation between level of training and total score (r = 0.82, p < 0.01). There was a significant increase in scores in the cutting and suturing task over the 2-year period (p < 0.01). Transferring skills did not improve significantly (p = 0.11). CONCLUSIONS: Performance in the simulator improved over residency training and was correlated highly with postgraduate year. This simulator model is a valuable teaching tool for training and evaluation of basic laparoscopic tasks in laparoscopic surgery. PMID- 10459331 TI - Does percutaneous fine-needle aspiration biopsy aid in the diagnosis and surgical management of lung masses? AB - OBJECTIVES: To evaluate the ability of percutaneous, transthoracic fine-needle aspiration biopsy (FNAB) to correctly diagnose intrathoracic masses, to determine what complications were experienced and at what rate they occurred and to define more clearly the role of this technique in the surgical management of lung masses. DESIGN: A chart review. SETTING: Kingston General Hospital, Kingston, Ont., a tertiary care centre and university-affiliated teaching hospital. PATIENTS: One hundred and thirteen patients who underwent 117 percutaneous transthoracic FNABs between Jan. 1, 1991, and July 1, 1996. OUTCOME MEASURES: Patient demographics, size and location of the lesion, diagnostic result of FNAB, complications of the procedure, smoking history, number of needle passes made by the radiologist and results of any other available biopsy (i.e., through bronchoscopy, mediastinoscopy, pleuroscopy) and of surgical resection, as well clinical information pertaining to the disease state in patients with nondiagnostic or negative FNAB. RESULTS: Eighty-six masses (73.5%) were diagnosed as malignant, 31 biopsy specimens (26.5%) were either nondiagnostic or negative for malignancy. Of these specimens, 15 (48.4%) were subsequently shown to be cancer. In 64 biopsies (54.7%), the patient suffered pneumothorax, requiring hospitalization and chest tube insertion in 35 (29.9%) and 24 (20.5%) cases respectively. The size of the lesion was related to both the diagnostic accuracy and the incidence of pneumothorax. CONCLUSIONS: Percutaneous transthoracic FNAB should not be used routinely in the assessment of patients with lung masses who are medically fit to withstand surgery and are free of widespread disease. The results of FNAB do little to modify the course of surgical management in these patients. PMID- 10459332 TI - Mycotic aneurysm of the suprarenal aorta due to Streptococcus pneumoniae: case report and literature review. PMID- 10459333 TI - Endoscopic perforation of the rectum presenting initially as a change in voice. PMID- 10459334 TI - Successful treatment of Madura foot caused by Pseudallescheria boydii with Escherichia coli superinfection: a case report. PMID- 10459335 TI - The Janes Surgical Society. AB - The Janes Surgical Society was formed in 1953 by surgeons who had undertaken their surgical training during the tenure of Dr. Robert M. Janes, Professor of Surgery at the University of Toronto from 1947 to 1957. Over the next 35 years, this unique Canadian surgical society met annually at surgical centres on this continent and abroad as well as at certain resort areas from time to time. Members of the Janes Surgical Society could be found in major clinical and academic positions across the country from St. John's to Victoria. Their annual meetings served a dual purpose: they provided a forum for the exchange of scientific knowledge and ideas by the members; and they provided an opportunity for members and their wives to socialize and renew old friendships dating back to their residency days and to establish new relationships with surgeons and their wives from other countries, including the United States, United Kingdom, France and Sweden. Unfortunately, owing to death and retirements of its members, the Society can no longer hold scientific meetings and travel to distant centres. Its sole activity is now an annual dinner in Toronto, when members and their wives gather to recall the highlights and experiences in their lives that this unique surgical society provided. PMID- 10459336 TI - Gallstones resulting from abdominal shotgun wound. PMID- 10459337 TI - Guide for managing breast lumps. PMID- 10459338 TI - Karyotype studies in 18 ependymomas with literature review of 107 cases. AB - Cytogenetic studies from 17 pediatric ependymomas and 1 ependymoblastoma are presented. Eight tumors had abnormal karyotypes. Another 107 published cases of cytogenetic analyses from pediatric and adult ependymomas or ependymoblastomas were reviewed. Of the total 125 tumors, 83 (66%) had abnormal karyotypes, of which 24 had a sole autosomal abnormality. Approximately one third had monosomy 22 (-22) or breakpoint 22q11-13, with a higher incidence in adult (56%) versus pediatric (28%) tumors. Structural abnormalities of chromosomes 1, 6, and 17, and numerical abnormalities of 7, 9, 12, and 20, in particular, are also discussed. Although no primary cytogenetic abnormality is evident, these findings may provide direction for additional investigations regarding the classification of these tumors. PMID- 10459339 TI - Translocation (15;17)(q22;q21) as a secondary chromosomal abnormality in a case of acute monoblastic leukemia with tetrasomy 8. AB - We describe a case of acute monoblastic leukemia (AML M5a), originally presenting as granulocytic sarcoma of the testis, showing unusual cytogenetic abnormalities. Tetrasomy 8 (primary) and t(15;17)(q22;q21) (secondary) were detected in bone marrow cells 6 months post-diagnosis, both by routine karyotype analysis and by fluorescence in situ hybridization (FISH) studies on metaphases and interphase nuclei. Retrospectively, the same abnormalities were identified in the primary testicular lesion using interphase FISH. However, reverse transcriptase polymerase chain reaction (RT-PCR) did not reveal the presence of a classic PML/RAR alpha fusion transcript. To the best of our knowledge, this is the first case to be reported in the literature of AML showing tetrasomy 8 in combination with secondary t(15;17). PMID- 10459340 TI - AgNOR of human interphase cells in relation to acrocentric chromosomes. AB - Using simultaneous detection of fluorescence in situ hybridization (FISH) to acrocentric chromosome centromeres and argyrophilic nucleolar organizer regions (AgNOR), we investigated the number of AgNOR and involvement pattern of acrocentric chromosomes in the nucleoli in various types of human interphase cells. The number of AgNOR of normal gastric mucosal epithelial cells was 2.27 +/ 1.18 and was higher than that of lymphocytes (1.08 +/- 0.28) and lower than that of gastric cancer (7.76 +/- 3.21). The number of acrocentric chromosome centromere signals of normal gastric mucosal epithelial cells was higher than that of normal leukocytes (P < 0.000), and lower than that of gastric cancer (P < 0.000). The acrocentric chromosome centromere signals in the lymphocytes and neutrophils were only half of that expected for diploid cells, perhaps related to acrocentric chromosome association. The proportion of acrocentric chromosomes attached to AgNOR in gastric cancer (0.88 +/- 0.22) was significantly higher than that of normal gastric mucosal epithelial cells (0.72 +/- 0.35, P = 0.007). In conclusion, acrocentric chromosome association appears to be present in circulating leukocytes even in interphase. The number of AgNORs and proportion of acrocentric chromosomes involved in AgNORs in human interphase cells may vary according to cell types. This could play a significant role in rDNA transcription and determination of cell phenotype, including malignant change. PMID- 10459341 TI - Is a duplication of 14q32 a new recurrent chromosomal alteration in B-cell non Hodgkin lymphoma? AB - Identification of clonal chromosomal abnormalities involving 14q32 and its association with specific histological subtypes of non-Hodgkin lymphoma (NHL) has provided substantial insight to the genetic events leading to the disease. However, in some cases with inferior morphology of tumor cell chromosomes, the additional segment on chromosome 14 remains unidentified by cytogenetic banding techniques alone. To elucidate the origin of the additional chromosomal segment and to correlate the newly determined alterations with histology, metaphases from 15 NHL patients with add(14)(q32) were examined using fluorescence in situ hybridization (FISH) techniques after cytogenetic analysis had been performed. We found the duplication of 14q involving the q32 region in 6 cases with a dup(14) (q32) in 4 cases and a dup(14)(q24q32) in 2 cases. In 8 cases, FISH unveiled known NHL associated translocations; a t(14;18)(q32;q21) in 4 cases, a t(11;14)(q13;q32) in 2 cases, a t(8;14)(q24;q32) and a t(9;14)(p13;q32) in 1 case each. We also noted a t(14;17)(q32;q21) in 1 case. The use of FISH was a valuable asset in determining the origin of the additional material on chromosome 14q32, and helped resolve a group of B-cell NHLs with involvement of a duplicated 14q32 region. PMID- 10459342 TI - Novel chromosome 16 abnormality--der(16)del(16) (q13)t(16;21)(p11.2;q22)- associated with acute myeloid leukemia. AB - Inversion of chromosome 16 is a common feature of acute myeloid leukemia (AML) M4, while t(16;21), although also associated with AML, appears to be a separate entity. We present a patient with myelodysplastic syndrome (MDS) who transformed to AML-M1. The karyotype was normal at diagnosis; at 15 months, hematological evidence of transformation was present, and repeat cytogenetics showed a novel rearrangement of one chromosome 16. Two breaks had occurred; one in the short arm at 16p11, with translocation of the segment distal to this onto chromosome 21q, and the other in the long arm at 16q22 with subsequent deletion of the segment from 16q22-->qter. Fluorescence in situ hybridization (FISH) confirmed the abnormalities detected by cytogenetics and excluded involvement of the AML1 gene on 21q22. While the 16q22 breakpoint was at the usual site for the inv(16), the 16p11 was not. The patient is more characteristic of t(16;21) than inv(16), and adds to the spectrum of chromosome 16 abnormalities in AML. PMID- 10459343 TI - Detection of translocation 8;21 on interphase cells from acute myelocytic leukemia by fluorescence in situ hybridization and its clinical application. AB - To detect a translocation (8;21)(q22;q22) in interphase cells by fluorescence in situ hybridization (FISH), we investigated three probe combinations: single-color hybridization with two cosmid probes (cY8 and cY3), single-color hybridization with four cosmid probes (cY8, cY3, cY107, and cYR4), and dual-color hybridization with two cosmid probes (cY107 and cYR4) from the AML1 gene flanking or overlapping the breakpoint region. Over 95% of nuclei gave sufficient signals in all three probe combinations, and the detection rates were not statistically different among them. Among 18 patients examined at diagnosis, 12 with t(8;21) were also monitored for the number of residual leukemic cells after chemotherapy or bone marrow transplantation (BMT). There were some discrepancies between morphology and genetic (especially FISH) results at partial, or even complete remission. As leukemic cells with t(8;21) can maturate, morphological assessment alone is insufficient to evaluate the residual leukemic cells. Interphase FISH provided more precise information about the clinical status of patients with an 8;21 translocation after treatment. PMID- 10459344 TI - Relation between cytogenetic characteristics of two human colonic adenocarcinoma cell lines and their ability to grow locally or metastasize or both: an experimental study in the nude mouse. AB - This study was aimed at elucidating the relation between the cytogenetic characteristics and the invasive ability of two human colonic adenocarcinoma cells lines, HT29 and CaCO2. These two cell lines have very different tumorigenic and metastatic capacities after intrasplenic injection into nude mice: high for HT29 and relatively weak for CaCO2. At the time of injection, cytogenetic studies of the two cell lines revealed shared abnormalities: paratriploidy with seven common extra chromosomes or chromosome regions and specific particularities. In HT29 cells, we observed a large marker of unknown origin, an isochromosome i(11)(q10) and 5, 12, 13, 15, 19, and (19q+) supernumerary chromosomes, and, finally, the absence of one chromosome 16. In CaCO2 cells, we observed a chromosome 1-derived marker with q24-31 duplication, 12q and 16 supernumerary chromosomes, and a der(16) marker. The most striking difference between the karyotypes of these two cell lines concerned chromosome 16 (under- and overexpressed in HT29 and CaCO2 cells, respectively), overexpression of chromosomes 13, 15, and 19 in HT29 cells, and the relative loss of 12p in CaCO2 cells. Although some differences may be due to the intrinsic characteristics of the stem line, the establishment of specific cytogenetic abnormalities points out the role of many regions of the genome in tumorigenic and metastatic capacities of malignant cells. PMID- 10459345 TI - Deletion 5q31 in patients with stable, melphalan-treated multiple myeloma. AB - The risk of myelodysplastic syndrome (MDS) or acute myeloblastic leukemia (AML) in patients with multiple myeloma has been estimated to be 10-20% after 10 years. Most myeloma patients develop MDS/AML after 3-4 years of treatment with alkylating agents, mainly melphalan; chromosomes 5 and 7 are most frequently involved. We studied 14 patients with myeloma by fluorescence in situ hybridization (FISH) with a probe to 5q31 (the critical area of deletion on chromosome 5) to verify whether deletion of 5q31 occurs during the course of stable, uncomplicated myeloma, and to assess the clinical importance of this abnormality. We found 2 patients (14%) with deletion of 5q31 in 30-40% of their peripheral white blood cells. One patient with this deletion received a high cumulative amount of melphalan, and the other patient was treated with multiple alkylating agents, including melphalan. In these patients, no clinical or laboratory evidence of transformation occurred 14 and 12 months after the finding of the aberration. These findings suggest that 5q-may occur months prior to the overt development of (t)-MDS/AML, and raise important concerns regarding the management of patients with this and similar aberrations, including modification of treatment and performance of cytogenetic evaluation prior to autologous or PSC transplantation. The clinical and biological implications of these findings should be evaluated in larger clinical and laboratory studies. PMID- 10459346 TI - Cytogenetic analysis of a primary salivary gland myoepithelioma. AB - Myoepithelioma, a rare benign salivary gland neoplasm, is a tumor composed entirely of myoepithelial cells. Unlike pleomorphic adenoma, these tumors lack any ductal epithelial differentiation, and manifest a minor stromal element. Previous cytogenetic and molecular genetic studies have mainly investigated pleomorphic adenomas and reported recurring specific chromosomal alterations at 8q12 and 12q13-q15 regions. The cell origin of these alterations, however, remains speculative. We report the cytogenetic analysis of a parotid myoepithelioma and discuss the putative origin for the cells with cytogenetic alterations. Our analysis shows 12q12 involved in a translocation with a previously unreported partner (1q), and nonrandom del(9)(q22.1q22.3) and del(13)(q12q22). Our results indicate that the myoepithelial cell is the source of those cells with chromosomal alterations, and that myoepithelioma shares 12q alterations reported in a subset of pleomorphic adenomas. PMID- 10459347 TI - Cytogenetic divergence of the same blastic clone in transformed chronic granulocytic leukemia: no effect on morphologic and immunologic features. AB - A 19-year-old man with Ph-positive chronic granulocytic leukemia developed lymphoblastic transformation. Cytogenetic evolution was observed, with an abnormal clone showing i(17q) together with the t(9;22). Chronic phase of the chronic granulocytic leukemia were re-established with systemic chemotherapy, which also led to disappearance of the clone with i(17q). However, the acute lymphoblastic leukemia relapsed after 6 weeks, with the emergence of a phenotypically and genetically identical but cytogenetically distinctive clone. Our findings suggest that cytogenetic evolution in transformed chronic granulocytic leukemia reflects only the instability of the blastic clones, and may not determine its phenotypic differentiation. PMID- 10459348 TI - Classification of IVS1-10T-->C as a polymorphism of BRCA1. AB - Mutations inactivating the tumor suppressor gene BRCA1 may be responsible for disease for up to 80% of familial ovarian cancer cases. In this syndrome, tumorigenesis classically initiates from an inherited mutation in one allele followed by somatic deletion of the normal allele. Sequencing of BRCA1 amplified from genomic DNA of lymphocytes and microdissected ovarian tumor cells of a familial ovarian cancer patient revealed three, rare heterozygous DNA variations (2418delA, 233G-->A, and IVS1-10T-->C) in both tumor and constitutional (lymphocyte) DNA. Thus, both copies of BRCA1 were retained in tumor. Haplotype analysis of the patient and four siblings assigned 2418delA to one copy of BRCA1 and 233G-->A and IVS1-10T-->C to the other. The DNA change, 2418delA, is considered a mutation that inactivated one BRCA1 allele because it caused a frameshift and generation of a premature stop codon, resulting in synthesis of a truncated peptide as evidenced by an in vitro protein truncation test. The DNA variation, 233G-->A, does not result in an amino acid change, and is considered a benign polymorphism. IVS1-10T-->C is a unique BRCA1 change that occurs in the last nucleotide of a consensus sequence for a branch site critical for RNA splicing. Therefore, we investigated whether IVS1-10T-->C deleteriously affected BRCA1 splicing or expression, and thereby inactivated the other BRCA1 allele. Using the technique of reverse transcription-polymerase chain reaction (PCR) with RNA isolated from lymphoid cell lines of the patient and of controls, no evidence was found that IVS1-10TC abnormally disrupted mRNA splicing or caused the absence of BRCA1 mRNA. Thus, IVS1-10T-->C is not harmful to BRCA1 function, and is classified a benign polymorphism. Retention of the normal BRCA1 allele in the tumor with the heterozygous germline BRCA1 mutation, 2418delA, indicated that mutational inactivation of both BRCA1 alleles was not required for tumorigenesis. It is possible that the normal allele may be functionally inactivated by a nonmutational mechanism. PMID- 10459349 TI - Allelic loss of the NF1 gene in NF1-associated plexiform neurofibromas. AB - Neurofibromatosis 1 (NF1) is an autosomal dominant disorder with a complex variety of clinical symptoms. Genetic alteration of the NF1 gene on 17q11.2 is the disease. Neurofibromas of the peripheral nervous system are one main manifestation. A variant of neurofibroma is the plexiform neurofibroma which can be found in about 30% of NF1-patients, often causing severe clinical symptoms. In this study, we examined 14 such tumors from 10 NF1-patients for allele loss of the NF1 gene (LOH: loss of heterozygosity) using four intragenic polymorphic markers. Loss of heterozygosity was found in eight tumors from five patients, and suspected in one additional tumor from another patient. This finding suggests that loss of the second allele, and thus inactivation of both alleles of the NF1 gene, is associated with the development of plexiform neurofibromas. The 14 plexiform neufibromas were also examined for mutation in the TP53 gene by screening exons 5 through 8 using temperature gradient gel electrophoresis. No mutation was found in any of the tumors. PMID- 10459350 TI - Consistent fusion of MOZ and TIF2 in AML with inv(8)(p11q13). AB - We have recently cloned the inv(8)(p11q13) in a patient with acute myeloid leukemia (AML), and demonstrated a fusion between the MOZ and TIF2 genes at 8p11 and 8q13, respectively. We have partially characterized a further case of AML with the same karyotypic abnormality. Rearrangements were detected by Southern blotting with a TIF2 probe that was close to the breakpoint in the original inv(8) case and with a MOZ probe corresponding to the breakpoint cluster region in the t(8;16) (p11;p13). These findings indicate the existence of breakpoint cluster regions within both genes and demonstrate that the MOZ-TIF2 fusion is consistently associated with the inv(8)(p11q13). PMID- 10459351 TI - Cytogenetic abnormalities correlate with the plasma cell labeling index and extent of bone marrow involvement in myeloma. AB - Chromosomal abnormalities have biologic and prognostic significance in multiple myeloma, especially among patients with relapsed disease. We report the relationship between chromosomal abnormalities and known prognostic factors such as plasma cell labeling index (PCLI) and bone marrow plasma cell involvement in 75 consecutive patients undergoing autologous stem cell transplantation for relapsed or refractory myeloma. Thirty of 70 patients (43%) had a chromosomally abnormal clone in their bone marrow, and in most cases the karyotype was complex (> 3 abnormalities). Patients with an abnormal clone on cytogenetic analysis had a higher PCLI (median, 1.4) than patients with a normal karyotype (0.2) (P < 0.001). Bone marrow plasma cell percentage also differed: median 48% versus 20%, respectively (P < 0.001). The PCLI and bone marrow plasma cell percentage correlated positively with the percentage of abnormal metaphases on conventional cytogenetic analysis: rho 0.60 (P < 0.001) and 0.46 (P < 0.001), respectively. We categorized patients into those with 20% or more abnormal metaphases, less than 20% abnormal metaphases, and only normal metaphases. The median PCLI values were 3.3, 1.1, and 0.3, respectively (P < 0.001). The bone marrow plasma cell percentage median values were 62%, 40%, and 25%, respectively (P = 0.003). Chromosomal abnormalities may offer a proliferative advantage to the neoplastic plasma cell, thereby leading to an unfavorable outcome. PMID- 10459352 TI - SRY mutation and tumor formation on the gonads of XP pure gonadal dysgenesis patients. AB - We report three patients with XY pure gonadal dysgenesis. Two of these patients developed gonadoblastoma and associated dysgerminoma. Molecular analyses were undertaken to investigate the relationship between the formation of these tumors and Y chromosome aberrations. Deletion analyses were performed by polymerase chain reaction (PCR) amplification of Y chromosome-specific DNA sequences (PABY, SRY, DYS250, DYS254, and DYZ1). A cryptic deletion of the short arm of the Y chromosome that included the PABY, SRY, DYS250, and DYS254 loci was observed in one of the patients (22-years-old) with an associated tumor. In the other two patients who did not demonstrate such a deletion, the sequence of the SRY open reading frame was determined by the dideoxynucleotide method. Two nucleotide substitutions followed by a seven nucleotide deletion were observed in the 3' end of HMG (high mobility group)-box in the other patient (15-years-old) with an associated tumor. The patient (22-years-old) without an associated tumor did not have the cryptic deletion or mutation of SRY. A Y chromosome specific sequence (DYZ1) was demonstrated by PCR amplification of microdissected tumor tissues from these two patients. These results suggest that SRY may play a role in the formation of gonadal tumors, especially dysgerminoma. PMID- 10459353 TI - Unusual clinical course and acquisition of del(11)(q23) in second lymphatic blastic phase of a Ph-positive chronic myeloid leukemia. AB - We describe unusual clinical and cytogenetic findings of a 29-year-old female with a Philadelphia chromosome (Ph)-positive chronic myeloid leukemia (CML), who showed a mosaic of apparently normal cells and cells bearing the classical t(9;22)(q34;q11) during the first lymphatic blastic phase (BP). The second lymphatic BP developed 10 years later. In addition to the t(9;22), which was detected in all metaphases, a del(11)(q23) was identified as a subclonal change in 4 of 25 metaphases. Fluorescence in situ hybridization (FISH) analysis using a chromosome 11-specific library probe and a probe covering the breakpoint cluster region of the MLL gene revealed hybridization signals of both probes on the normal and the deleted chromosome 11, indicating that the breakpoint on chromosome 11 occurred telomerically to the breakpoint cluster region of the MLL gene. Chemotherapeutic treatment resulted in reconstitution of the chronic phase with persistence of the Ph translocation as the sole chromosomal abnormality. PMID- 10459354 TI - Dicentric translocation (9;12) presenting as refractory Philadelphia chromosome positive acute B-cell lymphoblastic leukemia. AB - A 66-year-old Caucasian woman presented with a Philadelphia chromosome positive common B-cell acute lymphoblastic leukemia and a dic(9;12) involving the der(9)t(9;22), a rearrangement so far not observed in acute lymphoblastic leukemia. The patient was treated for the acute lymphoblastic leukemia, but showed refractory disease and died 6 months after initial diagnosis. This case suggests that, in the combination of t(9;22) and dic(9;12), the known poor prognostic feature of t(9;22) in acute lymphoblastic leukemia may outweigh the favorable outcome reported in patients with dic(9;12). PMID- 10459355 TI - 11q13 is a cytogenetically promiscuous site in hematologic malignancies. AB - 11q13 translocation has been described in mantle cell lymphoma in the form of t(11;14) (q13;q32), with rearrangement and over-expression of the cyclin D1 gene. Recently, an association between 11q13 and acute myeloid leukemia is recognized. We describe the occurrence of 11q13 translocations in both acute leukemias and myelodysplastic syndrome, and suggest that other genetic mechanisms unrelated to cyclin D1 may be involved in the tumorigensis. Furthermore, 11q13 appears to be a cytogenetically promiscuous site involved in reciprocal translocations with different chromosomes in both myeloid and lymphoid malignancies. PMID- 10459356 TI - Early onset of gastric carcinoma and constitutional deletion of 18p. AB - We report on the association of a gastric carcinoma and a constitutional deletion of the short arm of chromosome 18 in a 14-year-old patient. The phenotype of the patient, including microcephaly, ptosis, micrognathia, tetralogy of Fallot, and mental retardation, fits well with previously reported cases of del(18p); she also showed a positive serology against Helicobacter pylori. The comparison of the alleles of polymorphic loci located on the short arm of chromosome 18 between the patient and her parents showed a maternal origin of the abnormal chromosome. Loss of heterozygosity (LOH) for loci located in the long arm of chromosome 18 is a frequent event in gastric carcinomas; it was observed in the tumoral mass of our patient and again, the alleles lost were of maternal origin. We postulate that the constitutional chromosomal abnormality may have favored the loss of the abnormal chromosome in some cells and that the loss of the deleted chromosome 18 (demonstrated by LOH for this chromosome in the tumoral mass) has been an early step in the pathogenesis of the gastric carcinoma of our patient with Helicobacter pylori infection acting as a cofactor. PMID- 10459357 TI - p190 BCR-ABL rearrangement in chronic myeloid leukemia and acute lymphoblastic leukemia. AB - A minority of chronic myeloid leukemia (CML) cases have breakpoint in the minor cluster region (m-bcr) of the BCR-ABL fusion gene. We report a patient with Ph positive acute lymphoblastic leukemia and m-bcr breakpoint at diagnosis. The patient was treated with chemotherapy followed by an autologous peripheral blood stem cell transplantation, achieving a clinical and hematological complete remission but with persistence of the Philadelphia chromosome. One year later, she developed leukocytosis with a blood picture consistent with CML. She was treated with hydroxyurea and interferon alpha with no response. This is the second case of m-bcr CML reported presenting with features of lymphoid blast crisis or acute lymphoblastic leukemia. PMID- 10459358 TI - Atypical chromosome abnormalities in a renal oncocytoma. PMID- 10459359 TI - Computerized prescribing: steps to improve therapy. PMID- 10459360 TI - Pulmonary infiltrates in an elderly man. AB - An 80-year-old man presented with subjective fever, chronic cough occasionally producing scant yellow sputum, retrosternal pleuritic pain, and dyspnea on walking one block. Since symptom onset three months earlier, he had lost 20 pounds; he had had two loose stools a day, fatigue, malaise, and anorexia but not hemoptysis, nausea, vomiting, hematemesis, hematochezia, or melena. He denied paroxysmal nocturnal dyspnea or orthopnea. As far as could be ascertained, he not recently been exposed to tuberculosis or any other infectious disease. He had previously been seen at another clinic and had completed a 10-day trial of erythromycin (500 mg p.o. q12 h) without apparent change in symptoms. PMID- 10459361 TI - Supraventricular tachycardia in a man with unstable angina. PMID- 10459362 TI - The new vaccine for Lyme disease. PMID- 10459363 TI - Hemochromatosis: a "simple" genetic trait. AB - With the discovery of the causative gene, the disorder stands revealed as America's single most common mendelian disease. Unlike other genetic diseases, it is already curable. Indeed, genetic screening makes it potentially preventable. Yet a finding of disease-related genotype can also lead to stigmatization. Hemochromatosis therefore presents the issues surrounding genetic testing in especially stark form. PMID- 10459364 TI - Updating the approach to Crohn's disease. AB - Recognizing clinical and pathologic scenarios, rather than ordering definitive tests, is still the basis for diagnosis of Crohn's disease. Steroids, salicylates, immunomodulating agents, and surgery remain the focus of treatment. The advent of infliximab, however, offers a new therapeutic option for patients with refractory disease. PMID- 10459365 TI - Hormone replacement therapy: reassessing the risks and benefits. AB - Although the association between postmenopausal hormone use and breast cancer has not been clarified, there is nothing indefinite about the survival advantage conferred by hormone replacement therapy. Cardiovascular and fracture-related deaths so outnumber breast cancer deaths that even women with family histories of the latter are likely to benefit from low-potency estrogen replacement. PMID- 10459366 TI - Preventing falls in older adults. AB - For many older adults, falls are among the most ominous signs of aging, provoking fears of debilitating injury and loss of independence. Since most falls result from a complex interplay of intrinsic and extrinsic factors, environmental as well as physical challenges must be assessed. Some of the current strategies for preventing falls are reviewed. PMID- 10459367 TI - Peer-reviewed journals and quality. PMID- 10459368 TI - Judging the value of population-based disease management. PMID- 10459369 TI - Health insurance, health reform, and outpatient mental health treatment: who benefits? AB - This research examines how extending health insurance coverage to the previously uninsured impacts outpatient mental health treatment use among adults with different needs. Using data from the Epidemiologic Catchment Area Study and the 1987 National Medical Expenditure Survey, I develop simulations based on estimates of treatment demand. I find that insurance substantially increases demand by the mentally ill, but increased coverage alone cannot meet their treatment needs. Those in better mental health account for significant proportions of additional demand when coverage is expanded. Policies intended to increase access to mental health treatment among targeted groups should carefully consider the costs of increased use by other people. PMID- 10459370 TI - Episodes of mental health and substance abuse treatment under a managed behavioral health care carve-out. AB - Little is known about the effect of a managed behavioral health care (MBHC) carve out on treatment episodes for a mental health/substance abuse (MHSA) condition. This study found adoption of a carve-out for Massachusetts state employees associated with a dramatic drop in total MHSA costs per episode (particularly for individuals with certain severe MHSA conditions). The carve-out also was associated with a shift away from the use of facility care toward the use of outpatient care for enrollees with a diagnosis of unipolar depression. PMID- 10459371 TI - Health insurance, the quantity and quality of prenatal care, and infant health. AB - This paper presents a comprehensive analysis of the relationship between Medicaid, infant health, and the quantity and quality of prenatal care using data from the 1988 National Maternal and Infant Health Survey (NMIHS). This integrated approach provides a more complete picture of the effect of Medicaid and its avenues of influence, and is less likely to lead to spurious findings. The results indicate that there was no statistically significant relationship between insurance status and birth weight holding constant other observed characteristics, although there was some evidence that uninsured women and Medicaid recipients received less prenatal care than did privately insured women. Differences in prenatal care utilization, however, were small. In addition, there was no evidence that uninsured women or Medicaid recipients received lower quality prenatal care than privately insured women did even in a period prior to implementation of state programs aimed at ensuring high-quality care. The results of this paper raise questions about the efficacy of the current public health response to poor infant health that relies on expanding insurance coverage and enriched prenatal care programs. PMID- 10459372 TI - Small group reform and insurance provision by small firms, 1989-1995. AB - Since 1989, states have enacted legislation to dismantle barriers facing small businesses that wish to purchase health insurance. Using data on the insurance offerings of 2,472 small firms (one to 49 employees) observed from 1989 to 1995, we assess whether state reforms encouraged more small firms to sponsor health benefits. We find that small group reforms did not spur uninsured firms to offer insurance. Firms without health insurance say that the high price of coverage is still the major barrier they face to offering a plan. Our findings suggest that the small group reforms within the 1996 Health Insurance Portability and Accountability Act are not likely to have an effect on the small group market. Most states already had implemented measures similar to those found in the act, and not much changed. PMID- 10459373 TI - The Federal Employees Health Benefits Plan: implications for medicare reform. AB - This paper suggests that the Federal Employees Health Benefits Program (FEHBP) is perhaps a model for Medicare reform. First, we introduce the FEHBP and describe important features, such as the method for determining the government's premium contribution. Second, we examine the cost performance of the FEHBP program, and conclude that the FEHBP has out-performed private health insurance programs and Medicare in its ability to control costs. Third, we discuss the problem of adverse selection in the FEHBP. We conclude that the FEHBP has experienced some selection problems, but not enough to prevent it from offering a wide variety of choices without standardized benefits or direct risk adjustment. For a demonstration of competitive pricing in Medicare, the fourth section compares the FEHBP to two models of Medicare reform: "FEHBP for Medicare," proposed by Butler and Moffit; and the "Denver design." PMID- 10459374 TI - Population choice and variable selection in the estimation and application of risk models. AB - This paper shows that risk adjustment models based on demographic and employment variables are not easily transferable from one population to the next, and that administrative variables are not useful in predicting medical expenditures. We found statistically significant differences between models built on populations of employees from a single employer enrolled in two different health plans, and between models built on populations of enrollees from a single health plan employed at two different companies. Employment-based variables (e.g., length of employment) had little predictive power in any of these risk models. We conclude that policymakers should be careful in applying risk models across populations, and that future versions of risk models for use within large employers need not include employment-based variables. PMID- 10459375 TI - The effect of methodological differences in two survey's estimates of the percentage of employers sponsoring health insurance. AB - Two large surveys on employer-sponsored health insurance produced different estimates of the percentage of employers offering insurance to their employees in 1993. These differences occurred despite major similarities in the surveys' purpose and design. In this paper, five survey design factors are assessed. Estimates from the second survey were recomputed to eliminate cases not included in the first survey. Survey estimates were no longer significantly different when cases were removed because establishments had moved, were single-employee establishments on the sample frame, were classified as completed only in the second survey, or when poststratification adjustments in the weighting used only in the second survey were eliminated. Based on a comparison of 449 cases that responded in both surveys, changes in the wording of questions also probably contributed to the difference in survey estimates. These results indicate that estimates from these types of surveys are very sensitive to differing designs. PMID- 10459376 TI - The design of Healthcare for Communities: a study of health care delivery for alcohol, drug abuse, and mental health conditions. AB - There is a shortage of data to inform policy debates about the quickly changing health care system. This paper describes Healthcare for Communities (HCC), a component of the Robert Wood Johnson Foundation's Health Tracking Initiative that was designed to fill this gap for alcohol, drug abuse, and mental health care. HCC bridges clinical perspectives and economic/policy research approaches, links data at market, service delivery, and individual levels, and features a household survey of nearly 9,600 individuals with an employer follow-back survey. Public use files will be available in late 1999. PMID- 10459377 TI - CDC says upcoming doses of rotavirus vaccine should be delayed. PMID- 10459378 TI - Elimination of thimerosal from vaccines set as goal. PMID- 10459379 TI - Integrating pharmacy technicians into the health care team. PMID- 10459380 TI - Creatine. PMID- 10459381 TI - Harvey A. K. Whitney Lecture. At center stage: pharmacy in the next century. PMID- 10459382 TI - Accuracy of International Normalized Ratio determined by portable whole-blood coagulation monitor versus a central laboratory. PMID- 10459383 TI - Trainees' activities and experiences after a clinical pharmacy dialysis traineeship. PMID- 10459384 TI - Stability of tirofiban hydrochloride in three commonly used i.v. solutions and polyvinyl chloride administration sets. PMID- 10459385 TI - Applied pharmacoeconomics: evaluation and use of pharmacoeconomic data from the literature. AB - The evaluation and use of published pharmacoeconomic data are discussed. The pharmacoeconomic literature is a vast and powerful source of information for pharmacists and others who must make decisions about services and products. Published studies can provide data quickly and inexpensively, but they may not necessarily meet criteria for quality and may not be generalizable to a specific situation. Guidelines are available that are intended to standardize study methods, minimize potential bias, and increase the comparability and credibility of data. Before relying on a published study, decision-makers should ensure that it meets criteria related to objective, perspective, pharmacoeconomic method, design, interventions, costs, outcomes, discounting, results, sensitivity analysis, extrapolation of conclusions, and sponsorship. A sound study can be used to support decisions in such diverse areas as individual patient treatment, formulary management, drug-use guideline development, disease management initiatives, and pharmaceutical service evaluation. A decision-maker may be able to use the results by applying them at face value, employing a sensitivity analysis, performing a meta-analysis, incorporating the data into a pharmacoeconomic model, or reproducing the study on a smaller scale. The selection of an application strategy should be based on the potential impact of the decision on the quality and cost of care, with more rigorous and complex strategies reserved for more extreme impacts. Published pharmacoeconomic data can help pharmacists make better and more informed decisions about pharmaceutical services and products. PMID- 10459386 TI - ASHP statement on pharmacist decision-making on assisted suicide. PMID- 10459388 TI - ASHP statement on the pharmacist's role in primary care. PMID- 10459387 TI - ASHP statement on confidentiality of patient health care information. PMID- 10459389 TI - Complementary lipid-lowering therapies. PMID- 10459390 TI - Perspective on ASHP's assisted-suicide policy. PMID- 10459391 TI - Paronychia of the great toes associated with protease inhibitors. PMID- 10459392 TI - Preliminary survey of pharmacists' use of the Internet. PMID- 10459393 TI - Sleep deprivation in depression: what do we know, where do we go? AB - Manipulations of the sleep-wake cycle, whether of duration (total or partial sleep deprivation [SD]) or timing (partial SD, phase advance), have profound and rapid effects on depressed mood in 60% of all diagnostic subgroups of affective disorders. Relapse after recovery sleep is less when patients are receiving medication; it may be prevented by co-administration of lithium, pindolol, serotonergic antidepressants, bright light, or a subsequent phase advance procedure. Diurnal and day-to-day mood variability predict both short-term response to SD and long-term response to antidepressant drug treatment. These mood patterns can be understood in terms of a "two-process model of mood regulation" based on the model well established for sleep regulation: the interaction of circadian and homeostatic processes. The therapeutic effect of SD is postulated to be linked to changes in disturbed circadian- and sleep-wake dependent phase relationships and concomitant increase of slow-wave-sleep pressure; additionally, SD-induced sleepiness may counteract the hyperarousal state in depression. This model has the advantage of providing a comprehensive theoretical framework and stringent protocols ("constant routine," "forced desynchrony") to dissect out specific disturbances. Many aspects tie in with current serotonergic receptor hypotheses of SD action. A treatment inducing euthymia in severely depressed patients within hours is an important therapeutic option that has come of age for clinical use. PMID- 10459394 TI - Regional brain activity during transient self-induced anxiety and anger in healthy adults. AB - BACKGROUND: Several studies have demonstrated that transient self-induced sadness activates anterior paralimbic structures. To further examine the specificity of these findings and the neural substrates involved in anger and anxiety, we studied the neural correlates of the induction of anxiety and anger in healthy adults. METHODS: We used H2(15)O and positron emission tomography (PET) to measure regional cerebral blood flow (rCBF) in 16 healthy adults during the induction of transient anxiety, anger, and neutral emotions. Subjects achieved differential emotions by recalling prior life events while viewing affect appropriate faces. RESULTS: Both the anxiety and anger conditions were associated with increased normalized rCBF in left inferior frontal and left temporal pole regions and decreased rCBF in right posterior temporal/parietal and right superior frontal cortex, compared to the neutral induction. Additionally, compared to neutral induction, anxiety was associated with increased rCBF in the left anterior cingulate and cuneus and decreased rCBF in right medial frontal cortex, while the anger induction was uniquely associated with increased rCBF in right temporal pole and thalamus. CONCLUSIONS: Self-generated transient states of anxiety and anger are associated with both overlapping and distinct regional brain activity patterns and provide a template for further dissection of specific components of normal and pathologic emotions. PMID- 10459395 TI - Anger in healthy men: a PET study using script-driven imagery. AB - BACKGROUND: An understanding of the neurobiological basis of normal emotional processing is useful in formulating hypotheses regarding the pathophysiology of psychiatric illnesses. METHODS: This study examined the mediating functional neuroanatomy of anger in eight healthy men. Narrative scripts were developed from autobiographical information to induce anger and neutral states. The subjects imagined the content of the narrative scripts to induce anger during positron emission tomography to measure normalized regional cerebral blood flow (rCBF). Psychophysiologic responses and subjective ratings of emotional state were measured for each condition. Statistical parametric maps were constructed to reflect the Anger versus Neutral state contrast. RESULTS: Anger was associated with activation of the left orbitofrontal cortex, right anterior cingulate cortex affective division, and bilateral anterior temporal poles. CONCLUSIONS: These results suggest that the subjective experience of anger is associated with rCBF increases in anterior paralimbic regions of the brain. PMID- 10459396 TI - Morphometry of the dorsal raphe nucleus serotonergic neurons in suicide victims. AB - BACKGROUND: The serotonin deficiency hypothesis of suicide has been important heuristically. Few studies have directly examined the brainstem dorsal raphe nucleus (DRN) serotonin neurons. We determined the number and morphometry of DRN serotonergic neurons in suicide victims (n = 7) compared to controls (n = 6). METHODS: Brainstems were collected at autopsy, fixed and cryoprotected. Tissue was sectioned, stained for Nissl and processed with an antiserum that cross reacts with tryptophan hydroxylase. All DRN neurons were identified, counted and analyzed every 1000 microns. Neuron morphometry was characterized by soma area (micron 2), sphericity, perimeter, length and density (neurons per mm3). RESULTS: Neuron number and density was higher in suicide victims (1,780 +/- 127 neurons/mm3) than controls (1,349 +/- 68). The DRN volume did not differ between groups (66 +/- 9 mm3 for controls vs. 67 +/- 5 mm3 for suicides). Mean neuronal area and sphericity did not differ between suicides and controls. The total number and the density of DRN neurons did not correlate with age. CONCLUSIONS: The finding of an increased number of neurons indicates that impaired serotonergic transmission found in association with serious suicide attempts is not due to fewer neurons. PMID- 10459397 TI - Decreased platelet peripheral-type benzodiazepine receptors in adolescent inpatients with repeated suicide attempts. AB - BACKGROUND: Peripheral-type benzodiazepine receptors (PBR) are responsible for mitochondrial cholesterol uptake, the rate limiting step of steroidiogenesis. They have been shown to be increased after acute stress, and decreased during exposure to chronic stressful conditions, and in patients with generalized anxiety disorder and post-traumatic stress disorder. In view of the proven connection between adolescent suicidal behavior and stress, we hypothesized that PBR may be decreased in the suicidal adolescent population. METHODS: We measured [3H] PK 11195 binding to platelet membrane in nine adolescent (age 13-20 years) inpatients with a history of at least three suicidal attempts and ten age-matched psychiatric inpatients with no history of suicide attempts. Suicidality was assessed with the Suicide Risk Scale (SRS), and symptom severity with the Beck Depression Inventory, State-Trait Anxiety Inventory (STAI), Overt Aggression Scale (OAS), and Impulsivity Scale (IS). RESULTS: Suicide Risk Scale scores were significantly higher in the suicidal group. The suicidal group showed a significant decrease in platelet PBR density (-35%) compared to the controls (p < 0.005). CONCLUSIONS: Our results of PBR depletion in adolescent suicide are in accordance with the findings in patients with generalized anxiety disorder and posttraumatic stress disorder and lend further support to the role of PBR in human response to chronic stress in adolescent suicide. PMID- 10459398 TI - Mood effects of 24-hour tryptophan depletion in healthy first-degree relatives of patients with affective disorders. AB - BACKGROUND: Acute tryptophan (TRP) depletion was evaluated in healthy volunteers with or without a family history of major affective disorder (FH+ versus FH-). METHODS: Twenty-seven subjects (16 FH+, 11 FH-) received 100 g of an amino acid mixture with and without TRP according to a placebo-controlled, double-blind cross-over design and a diet devoid of TRP for the next 24 hours. RESULTS: The ratio TRP/large neutral amino acids declined to 22% of baseline values after 6 hours, and increased during the night reaching 85% of baseline after 24 hours. Overall, after 6 hours, TRP depletion lead to a lowering of mood, but after 24 hours, these changes were no longer detected. Mood changes and gastrointestinal side effects were significantly more evident in FH+ subjects than in FH- subjects. CONCLUSIONS: Our data support the hypothesis that subjects with a positive family history for depression are predisposed to increased vulnerability to the adverse consequences of serotonergic imbalance. PMID- 10459399 TI - Tryptophan depletion and depressive vulnerability. AB - BACKGROUND: Rapid and transient depletion of tryptophan (TRP) causes a brief depressive relapse in most patients successfully treated with and taking selective serotonin reuptake inhibitors, but little change in drug-free, symptomatic depressed patients. This study investigates the effects of TRP depletion in drug-free subjects in clinical remission from a prior major depressive episode (MDE). METHODS: Twelve subjects with a prior MDE, currently in clinical remission and drug-free for at least 3 months (patients), and 12 healthy subjects without personal or family history of Axis I disorder (controls), received TRP depletion. The study was conducted in a double-blind, controlled [full (102-g) and quarter-strength (25 g) 15-amino acid drinks], crossover fashion. Behavioral ratings and plasma TRP levels were obtained prior to, during, and after testing. RESULTS: All subjects experienced significant depletion of plasma TRP on both test-drinks, showing a significant dose-response relation. Healthy control subjects had minimal mood changes, but patients had a depressive response of greater magnitude. CONCLUSIONS: In the context of prior TRP depletion studies with antidepressant-treated, and drug-free symptomatic depressed patients, these results suggest that depression may be caused not by an abnormality of 5-HT function, but by dysfunction of other systems or brain regions modulated by 5-HT. PMID- 10459400 TI - Prolactin secretion in depressed children. AB - BACKGROUND: Few studies have examined the involvement of the central dopaminergic system in the pathophysiology of mood disorders. The study of prolactin (PRL) secretion may be an informative indirect method for the assessment of the dopaminergic system in children with major depressive disorder (MDD). METHODS: Plasma PRL concentrations were measured at 20-min intervals over a 24-hr period in 40 pre-pubertal children with MDD, 18 with non-affective psychiatric disorders (PC), and 6 normal controls (NC). A subgroup of depressed children (n = 21) was restudied after recovery. RESULTS: There was no significant differences in either the amount or the pattern of PRL secretion between the MDD, PC, and NC groups. Children who recovered from their depression secreted less PRL during sleep and more while awake compared to when they were acutely depressed. CONCLUSIONS: Overall, there were no differences in baseline PRL secretion between children with MDD, NC and psychiatric control. These results suggest that the dopaminergic system as measured by baseline PRL blood levels is not compromised in children with MDD. PMID- 10459401 TI - The influence of prolactin response to d-fenfluramine on executive functioning in major depression. AB - BACKGROUND: The main purpose of this study was to assess the influence of serotonergic activity, as measured by prolactin response to d-fenfluramine, on executive functioning in major depression. METHODS: Forty-one persons between 22 and 71 years of age who met the DSM-IV criteria for major depressive episode were administered the Wisconsin Card Sorting Test (WCST) in computerized format, and d fenfluramine was administered orally. Postfenfluramine blood samples for ascertaining plasma prolactin levels were obtained. RESULTS: The key finding was that prolactin response was positively related to four out of five selected WCST variables. Also, increasing age was associated with decreasing WCST performance. There was no interaction between prolactin response and age, indicating that the effects of prolactin response on the WCST generalized across the age range examined. CONCLUSIONS: The overall pattern of results suggests that there may be a relationship between serotonergic activity and executive functioning in major depression. PMID- 10459402 TI - Pretreatment platelet 5-HT concentration predicts the short-term response to paroxetine in major depression. Grupo de Trastornos Afectivos. AB - BACKGROUND: A previous retrospective study revealed that a high pretreatment platelet serotonin (5-HT) concentration was associated with a low response to serotonergic antidepressants in drug-free major depressives. We have examined such a relationship in depressive patients treated with paroxetine. METHODS: Seventy-four drug-free major depressives (DSM-IV) were admitted to the study. Clinical ratings were performed and blood was drawn prior to the initiation of treatment and after 4 weeks of paroxetine (20 mg/day). The concentrations of 5 HT, 5-hydroxyindoleacetic acid, and tryptophan were determined in plasma and blood. RESULTS: Paroxetine treatment reduced platelet 5-HT to 17% of baseline after 4 weeks of treatment. Responder patients had a pretreatment platelet 5-HT concentration 22% lower than nonresponders (p < .035). Admission HAMD scores, plasma paroxetine concentration, or platelet 5-HT concentration at endpoint did not differ between responders and nonresponders. Yet, the response rate was 11% in patients with high pretreatment platelet 5-HT (> 900 ng/10(9) platelets) and 50% in those below that value (p < .004). CONCLUSIONS: These findings support that depressed patients with a high pretreatment platelet 5-HT concentration have a poor therapeutic outcome after treatment with a standard paroxetine dose. These differences may be related to the existence of molecular differences in the 5-HT transporter. PMID- 10459403 TI - Declarative and procedural memory in bipolar disorder. AB - BACKGROUND: Memory function is an important but under researched area for neuropsychological investigation in persons with bipolar disorder. Previous studies have reported cognitive deficits on tasks of declarative memory in bipolar patients in the euthymic state. METHODS: This study extended these findings by investigating declarative as well as procedural learning and memory in bipolar patients (with and without alcohol abuse) who were examined in the euthymic state. The California Verbal Learning Test, Star Mirror Tracing Task, Pursuit Rotor Task, American National Adult Reading Test, and the Vocabulary Subtest of the WAIS-R, were administered to bipolar patients and control subjects by researchers who were blind to the subject's group. RESULTS: Bipolar patients performed worse than control subjects on a measure of declarative memory (California Verbal Learning Test) but did not differ from the performance of control subjects on either of the two procedural learning tasks (Pursuit Rotor Task and Star Mirror Task). CONCLUSIONS: These results suggest disturbed function of temporal lobe, but not basal ganglia, structures in persons with bipolar disorder. PMID- 10459404 TI - Serum cholesterol, suicidal tendencies, impulsivity, aggression, and depression in adolescent psychiatric inpatients. AB - BACKGROUND: This study was undertaken to examine the relationship between serum cholesterol levels and suicidal behaviors in adolescent psychiatric inpatients. Any association between serum cholesterol and measures of suicidal behavior, impulsivity, aggression, anxiety, and depression was also examined. METHODS: Consecutive admissions (n = 152) to an adolescent psychiatric inpatient unit were assessed using measures of suicidal behavior, violence, impulsivity, and depression. Serum cholesterol was compared between those admitted for reasons of suicidal tendencies and those for other reasons. Correlation between serum cholesterol and measures of suicidal behavior, violence, impulsivity, and depression were examined. RESULTS: Serum cholesterol levels were significantly higher in adolescent patients who were currently suicidal than in nonsuicidal adolescents. Within the suicidal group, but not in the total inpatient group, serum cholesterol correlated negatively with the degree of suicidal behavior. No correlation between serum cholesterol levels and depression, violence, and impulsivity were detected. No significant differences were found in serum cholesterol levels between diagnoses or between suicidal and nonsuicidal patients within each diagnostic group. CONCLUSIONS: The association between cholesterol and suicidal tendencies remains complex and may depend on several variables within the population studied. Its usefulness as a biologic risk factor in clinical samples remains to be determined. PMID- 10459405 TI - Affective startle modulation in clinical depression: preliminary findings. AB - BACKGROUND: Modulation of the startle reflex by affective foreground stimuli was investigated in a group receiving inpatient treatment for major depressive episodes (n = 14) and an age and gender matched nondepressed group (n = 14). METHODS: Participants viewed 27 pleasant, neutral, and unpleasant pictures chosen from the International Affective Picture System. Acoustic startle probes were presented during picture viewing, and participants also rated the affective qualities of the pictures. RESULTS: While ratings of the pictures were largely similar between the depressed and nondepressed groups, they displayed dissimilar patterns of startle modulation. In the nondepressed group, blinks elicited during unpleasant pictures were significantly larger than during pleasant pictures, whereas the depressed group failed to show this effect. Analyses, which separated the depressed participants into moderate and severe groups based on Beck Depression Inventory scores, revealed that while the moderately depressed group also showed a normal pattern of startle modulation, the severely depressed showed potentiated startles during the pleasant pictures. CONCLUSIONS: These preliminary results suggest that severely depressed patients may respond to some pleasant stimuli as if they are aversive, possibly because such stimuli are seen as signals of frustrative nonreward. PMID- 10459406 TI - Cerebrospinal fluid somatostatin, mood, and cognition in multiple sclerosis. AB - BACKGROUND: Cerebrospinal fluid (CSF) somatostatin (SS) levels have been shown to be decreased in multiple sclerosis (MS) during relapse as well as in disorders characterized by depression or cognitive impairment. Since MS is often associated with depression and cognitive impairment, we examined both the effect of course of illness on CSF SS as well as the variance in SS attributable to associated features (e.g., depression or cognitive impairment). METHODS: Fifteen patients with chronic progressive MS participating in a 2-year cyclosporine trial underwent lumbar punctures for CSF SS at baseline and at 12 and 24 months. Additionally, patients were evaluated by neuropsychological testing, and physical disability and mood ratings. Baseline CSF SS levels were also obtained in a group of control subjects (n = 10). RESULTS: At baseline, CSF SS levels were lower in MS patients than control subjects (p < .001). Decreased CSF SS at 24 months was correlated with decreased cognitive performance on several measures and was best and significantly predicted by cognitive deterioration at 24 months. CONCLUSIONS: Our data support those from previous studies that found lower levels of CSF SS in MS during relapse and suggest that changes in CSF SS are related to the process responsible for diminished cognitive function in MS. PMID- 10459407 TI - Global variation in the frequencies of functionally different catechol-O methyltransferase alleles. AB - BACKGROUND: Catechol-O-methyltransferase (COMT) has been investigated as a candidate gene in many neurologic disorders involving catecholaminergic systems. The NlaIII restriction site polymorphism (RSP) at COMT is a G<-->A (site absent<- >site present) single nucleotide polymorphism (SNP) at nucleotide 322/472 (in the short or long mRNA) that results in a Val<-->Met polymorphism at amino acid 108/158 (in soluble or membrane-bound) COMT protein and different enzyme activity levels, high for Val, low for Met. COMT enzyme activity is known to vary among ethnic groups, presumably as a result of different population frequencies of these COMT alleles. We have undertaken a direct survey of allele frequencies of this polymorphism in a global sample of populations. METHODS: We typed 1314 individuals from 30 different populations using PCR of the relevant region followed by digestion with NlaIII and electrophoresis. RESULTS: The frequencies of the low activity allele (COMT*L, NlaIII site-present) vary significantly from 0.01 to 0.62. Europeans have nearly equal frequencies of the two alleles while the COMT*H allele is much more common in populations in all other parts of the world. Sequencing in nonhuman primates indicates that COMT*H is the ancestral allele in humans. CONCLUSIONS: This is the first global survey of the COMT*L and COMT*H allele frequencies, confirming and extending earlier studies to show significant world-wide variation. This is also the first study establishing the COMT*L allele as the derived allele unique to humans. Henceforth, in any population-based association studies of this polymorphism, the control allele frequencies should be in agreement with these published values for corresponding ethnic groups. PMID- 10459408 TI - Salivary prolactin as a marker for central serotonin turnover. AB - Central nervous system (CNS) serotonin deficits have been linked to many pathological behaviors in both human and nonhuman primates. The plasma prolactin response to fenfluramine has been widely used to assess CNS serotonin functioning in humans. Prolactin is also found as an integrated measure in saliva. We hypothesized that salivary prolactin concentrations would correlate positively with cerebrospinal fluid (CSF) 5-hydroxyindoleacetic acid (5-HIAA) in rhesus monkeys. Twenty-seven adult male and female rhesus macaques (Macaca mulatta) were sampled for concurrent saliva, blood, and CSF. Saliva and blood serum were assayed for prolactin concentrations, and CSF was assayed for 5-HIAA, homovanillic acid (HVA), and 3-methoxy-4-hydroxyphenylglycol (MHPG). Salivary prolactin concentrations were positively correlated with CSF 5-HIAA concentrations. No other relationships between any of the measures, including that between salivary prolactin and serum prolactin, were found to be statistically significant. These findings suggest the possibility of using salivary prolactin concentrations as an index of CNS serotonin turnover in humans. PMID- 10459410 TI - Lack of relationship between menstrual cycle phase and mood in a sample of women with rapid cycling bipolar disorder. AB - BACKGROUND: Women are overrepresented in samples of patients with rapid cycling bipolar disorder (RCBD). To explore whether menstrually related mood changes might account for this gender difference, we studied the relationship between menstrual cycle phase and mood in a sample of premenopausal women with rapid cycling bipolar disorder (RCBD). METHODS: Twenty-five women with RCBD completed daily self-rating forms indicating their mood and days of menstruation for a minimum of three months. The data were analyzed for each individual and for the group as a whole, categorically (depression, euthymia, and hypomania) and ordinally (0-100, with 0 being "most depressed ever felt" and 100 being "most manic"), with and without normalization of the menstrual cycle to a 28-day cycle. RESULTS: None of the group analyses showed a significant effect of menstrual cycle on mood. Although some women did exhibit significant relationships between menstrual cycle phase and categorical mood state, there was no consistent pattern to the relationship. CONCLUSIONS: There was no systematic relationship between menstrual cycle and mood in a sample of women with RCBD. PMID- 10459409 TI - Effects of catecholamine depletion with AMPT (alpha-methyl-para-tyrosine) in obsessive-compulsive disorder. AB - BACKGROUND: Several lines of evidence suggest that brain dopamine function may contribute to some obsessive-compulsive (OC) phenomena. The effects of catecholamine depletion were examined in drug-free patients with obsessive compulsive disorder (OCD). METHODS: The tyrosine hydroxylase inhibitor alpha methyl-para-tyrosine (AMPT) and diphenhydramine hydrochloride (placebo) were administered for three consecutive days, one week apart, to 6 drug-free adult OCD patients without a personal or family history of chronic tics, in a double-blind, randomized design. The effects of AMPT and placebo on OC, depression, anxiety and global clinical symptoms were assessed. RESULTS: AMPT produced no clinically or statistically significant change in any behavioral ratings, including OC symptom severity, compared with placebo. CONCLUSIONS: Acute reduction of catecholamine levels does not seem to affect OC symptoms in drug-free patients with OCD. Studies of catecholamine depletion with AMPT in patients with comorbid OCD and chronic tics may be of considerable neurobiological and clinical interest. PMID- 10459412 TI - Integrating clinical engineering and information systems. PMID- 10459411 TI - Neuroendocrine evidence for dopaminergic actions of hypericum extract (LI 160) in healthy volunteers. AB - BACKGROUND: We studied the effect of a single dose of a formulation of a methanolic extract of Hypericum perforatum (HP), also known as St. John's wort, on plasma concentrations of growth hormone (GH), prolactin (PRL), and cortisol (CORT) in 12 healthy male volunteers. METHODS: Subjects received 9 tablets of the finished product Jarsin 300 and placebo in a double-blind, balanced-order, cross over design. RESULTS: Following HP relative to placebo, there was a significant increase in plasma GH and a significant decrease in plasma PRL. Plasma CORT levels were unchanged. CONCLUSIONS: Taken together with data from animal experimental studies, the findings suggest that this dose of HP may increase some aspects of brain dopamine function in humans. PMID- 10459413 TI - Medical image transmission tops list of telemedicine applications. PMID- 10459414 TI - Engineering endpoints and PM intervals. PMID- 10459415 TI - SGNA on reuse of devices. PMID- 10459416 TI - Defibrillator testing. PMID- 10459417 TI - Engineering in software testing: statistical testing based on a usage model applied to medical device development. AB - When a population is too large for exhaustive study, as is the case for all possible uses of a software system, a statistically correct sample must be drawn as a basis for inferences about the population. A Markov chain usage model is an engineering formalism that represents the population of possible uses for which a product is to be tested. In statistical testing of software based on a Markov chain usage model, the rich body of analytical results available for Markov chains provides numerous insights that can be used in both product development and test planing. A usage model is based on specifications rather than code, so insights that result from model building can inform product decisions in the early stages of a project when the opportunity to prevent problems is the greatest. Statistical testing based on a usage model provides a sound scientific basis for quantifying the reliability of software. PMID- 10459418 TI - Leadership, patient safety, and Y2K: the AAMI Technology Management Committee. PMID- 10459419 TI - Risk management and medical devices. PMID- 10459420 TI - IEC 60601-1, the 3rd edition: what will change? PMID- 10459422 TI - On-line techniques for perfluorochemical vapor sampling and measurement. AB - The authors developed a compact gas sampling and perfluorochemical (PFC) measuring system for use in total and partial liquid ventilation systems, based on a precision two-thermistor thermal detector (TD). They describe the sensitivity and linearity of their on-line method for PFC analysis of expired gases and show how it may be used in partial liquid ventilation studies for determining PFC saturation and loss. Gas is sampled for a short time from a breathing circuit through a heated tube at a selectable point in the breathing cycle. Inspiration is sensed by a pressure transducer. The sample of gas is pulled into the heated (48 degrees C) thermistor chamber by suction and held there while the cooling effect of the vapor changes the thermistor temperature. Dry air in another chamber affects a second thermistor, and the difference of these responses is amplified. The raw signal is corrected for the effects of varying O2 levels by a fuel cell. This signal is sampled and held and displayed on a front panel display. Calibration is performed in percentage saturation at 37 degrees C using the PFC in use at that temperature, or another standard such as O2. In-vitro testing showed a linear response in the thermal detector device (R2 = 0.99) over the range of vapor pressures tested (0-14) mmHg) and was reproducible to within 3%. When electronically corrected for changes in O2 concentration, there was less than a 2% change in PFC saturation. The TD responses to CO2 (R2 = 0.99) and water vapor (R2 = 1.0) were linear and approximately equal and opposite over the normal operating ranges of expired gases. In-vivo results in rabbits showed a significant (R2 = 0.73; p < 0.01) correlation between the auto-sampler and manual collection modes for determination of PFC in expired gas. PMID- 10459423 TI - Dose monitoring in Partial Liquid Ventilation by infrared measurement of expired perfluorochemicals. AB - Patients undergoing Partial Liquid Ventilation (PLV) with the perfluorochemical liquid perflubron (PFB) continuously evaporate the drug from the lung during ventilatory expiration. In this study, two infrared (IR) devices, a modified industrial analyzer ("experimental prototype") and a custom-designed device suitable for use in a clinical environment ("clinical prototype"), were calibrated and validated on the bench to measure a range of PFB concentrations (CPFB) in a gas stream. PFB loss from the lung (area under the CPFB-vs-time curve) could be correlated during PLV simulation with changes in tidal volume, breathing rate, and variable CPFB-vs-time profiles. The two IR devices produced nearly identical measurements for the same CPFB standards (maximum deviation = 1.5%). The experimental IR prototype was tested in 17 anesthetized, paralyzed, and ventilated swine (42-53 kg) to quantify the total amount and rate of evaporate loss of PFB over 12 hours of PLV, both with and without periodic supplemental PFB doses. The residual PFB volumes in the animal lungs at the end of the study, as determined by a gravimetric postmortem lung method, were found to agree on average for all animals to within 10% of the residual PFB volume as predicted by the IR approach. Furthermore, the IR signal of CPFB does not appear to correlate with the absolute amount of PFB in the lungs, but may reflect the relative proportion of PFB-wetted airway and alveolar surface. The authors conclude that IR quantitation of PFB evaporative loss is acceptably accurate for extended periods of PLV and may be a useful tool in the clinic for PFB dose monitoring and maintenance, thereby helping to optimize PLV treatment. PMID- 10459424 TI - Dose maintenance for Partial Liquid Ventilation: passive heat-and-moisture exchangers. AB - Partial Liquid Ventilation (PLV), a promising method for the treatment of acute lung failure, has been evaluated in many animal studies. It has recently progressed to the point of controlled clinical trials in which patients of all ages on conventional mechanical ventilation (CMV) have their lungs substantially filled with a perfluorochemical (PFC) liquid, perflubron (PFB). During PLV, it is desirable to both maintain humidification and minimize the evaporation of PFB in order to maintain a desired dose in the lung and to reduce dose consumption and redosing effort. Heat-and-moisture exchangers (HMEs) have been used for years as a passive means of minimizing water vapor loss from the respiratory tract during CMV support of intensive care and surgical patients. In the current study, research was undertaken to leverage the operating principles of existing HMEs such that specialized "fluorophilic" HMEs (FHMEs), devices optimized for both water and PFB conservation, could be realized. A patient simulator (involving both water vapor and PFB vapor sources) was constructed and used in the in-vitro evaluation of various FHME concepts. Dose-retention efficiencies were determined with the aid of an infrared instrument and a digital thermohygrometer. Although no larger than commercial HMEs in terms of dead space (gas-occupying volume), efficient FHMEs resulted, offering less flow resistance (delta P) than their commercial counterparts. Additionally, the presence of PFB vapor did not appear to compromise the water-exchange efficiency of certain HME configurations. One promising FHME design was also tested in swine undergoing 12-hour PLV treatments. A mean conservation efficiency of 63% at an average tidal volume of 550 mL was shown, although somewhat lower efficiencies may result in adult patients because efficiency was found to trend downward with increasing tidal volume. The use of an FHME is expected to sustain dose levels in patients for longer periods with less frequent dosing and reduced dose consumption, saving treatment labor and cost. PMID- 10459425 TI - A closed rebreathing system for dose maintenance during Partial Liquid Ventilation. AB - Partial Liquid Ventilation (PLV), a treatment for acute respiratory failure in which the lungs are filled, either partially or to functional residual capacity (FRC), with perfluorochemical (PFC) liquid while the patient is on mechanical gas ventilation, has progressed to clinical trials using the PFC perflubron (PFB). Because gas expired during PLV is laden with PFB vapor, PFB is lost via evaporation, which increases dose consumption and necessitates periodic redosing. A device has been developed to minimize evaporative loss by confining PFC vapor to a gas volume breathed by the patient, which is isolated from the ventilator. This closed rebreathing system works with the ventilator such that after the lung is filled with PFB, the patient is connected to the rebreathing system, with breathing still "driven" by the ventilator. The rebreathing system consists of two gas circuits, or compartments, separated by a flexible bag (in a box) partition. One compartment is in gas communication with the lung, while the second communicates with the ventilator. The O2 level on the patient side is matched to that on the ventilator side by sensing gas concentrations and by feedback control of O2 introduction. Similarly, air is introduced into the patient side under pressure-based feedback control to maintain a constant gas volume. On inspiration, the ventilator delivers the tidal volume (breath) into the box surrounding the bag, which, in turn, is transmitted through the bag to the lung. On expiration, the process is reversed. Unidirectional circulation of gas in the rebreathing circuit is achieved via check valves, and expired CO2 is removed by a barium hydroxide lime cartridge. Airway humidification is maintained by captive water vapor in the system and water vapor from the CO2 absorber. It is recommended that flow, pressure, O2, and CO2 levels be monitored at the patient "Y," i.e., the proximal end of the endotracheal tube. Performance data from both in-vitro experiments and in-vivo PLV experiments in pigs are presented. The authors conclude that with the closed rebreathing system, the dose can be safely maintained with fewer redosing procedures, and an approximately 90% savings in dose is achieved. PMID- 10459426 TI - Postoperative care and monitoring. AB - Pneumonectomy remains one of the more risky and morbid procedures routinely performed by the thoracic surgeon. Patients are often frail, with poor underlying pulmonary function and minimal physiologic reserve. Attentive, meticulous postoperative care and monitoring are required to minimize the incidence and severity of complications. PMID- 10459427 TI - Morbidity and mortality after pneumonectomy. AB - Although advances in perioperative management have improved outcome following pneumonectomy, it remains a relatively high-risk thoracic surgical procedure. A 30-day mortality rate of 5% to 10% is reported in most modern series. A higher rate is expected for complex procedures and in the setting of pneumonectomy performed for benign disease. A number of risk factors predictive of poor outcome have been described and are reviewed in this article. Pneumonectomy is also associated with a higher complication rate (predominantly cardiopulmonary in nature) than lesser resections. PMID- 10459428 TI - Early complications. Cardiac complications. AB - Cardiopulmonary complications developing after pneumonectomy are described. Arrhythmias are seen in about 25% of patients undergoing pneumonectomy, and supraventricular tachyarrhythmias are common. To date, the efficacy of medical prophylaxis has not yet been defined. Postpneumonectomy heart failure shares a common cause with postpneumonectomy edema. Cardiac herniation with subsequent torsion is a rare but fatal complication after pneumonectomy. Early recognition and prophylaxis are always crucial. PMID- 10459429 TI - Early complications. Respiratory failure. AB - Pulmonary complications following thoracic surgery are common and associated with significant morbidity and mortality. Respiratory failure after pneumonectomy occurs in approximately 5% to 15% of cases and significantly increases patient mortality. Strategies for ventilator support are based on the nature of the underlying complication and the pathophysiology of respiratory failure. This article describes the cause and pathophysiology of respiratory failure and pulmonary embolus postpneumonectomy. Diagnosis, management, and innovative therapies are also reviewed. PMID- 10459430 TI - Early complications. Recurrent nerve palsy. AB - Unilateral vocal cord paralysis following pneumonectomy has profound effects on deglutition, phonation, protection of the airway, and generation of an adequate cough. If untreated in patients with limited pulmonary reserve, these detrimental effects can have devastating consequences. Several techniques are currently available which allow adequate medialization and rehabilitation of the paralyzed vocal cord. Adequate diagnosis and treatment can minimize the negative consequences associated with vocal cord paralysis following pneumonectomy. PMID- 10459431 TI - Early complications. Chylothorax. AB - Postpneumonectomy chylothorax is a very common but serious complication. Drainage of the pneumonectomy space, metabolic and nutritional support with TPN, and absolute enteral rest may lead to control of the leak. Failure of these measures to obtain a rapid resolution of the chyle losses should be followed by early surgical intervention in most instances in an effort to alleviate the chronic metabolic, nutritional, and immunological consequences of prolonged chyle losses. PMID- 10459432 TI - Early complications. Esophagopleural fistula. AB - Esophagopleural fistulae complicate the outcome of approximately 0.5% of pneumonectomies, regardless of whether performed for benign or malignant conditions. Early postoperative fistulae result from operative injury to the esophagus: both direct tears of the mucosa and devascularization with secondary necrosis have been documented. Late esophagopleural fistulae, diagnosed beyond the third postoperative month, are due to cancer recurrence or various inflammatory disorders. The usual presentation is empyema thoracis. Diagnosis is suggested by drainage of food particles or saliva, and the presence of yeast cells within the pleural fluid. Confirmation relies on direct opacification of the fistulous tract during opaque swallow studies. Treatment is initiated by clearance of empyema with either tube thoracostomy or Clagett window, and feeding gastrostomy or jejunostomy. PMID- 10459433 TI - Late complications. Late respiratory failure. AB - Late and progressive respiratory failure after pneumonectomy may result from a variety of causes. Non-specific causes include restrictive failure by loss of alveolar volume; pulmonary hypertension; initial disease recurrence (e.g. bronchogenic cancer, bronchiectasis); side-effects of radio- and chemotherapy; and benign or malignant pleural or pericardial effusions. Acute or subacute conditions are congestive or ischemic heart failure, pulmonary embolism, and pneumonia. Two causes are specific, benign, and curable: the postpnemonectomy syndrome and the platypneaorthodeoxia syndrome. The latter is related to a right to-left interatrial shunt through a reopened patent foramen ovale. The hemodynamic and anatomical mechanisms are analyzed through an exhaustive review of the literature, together with the particular clinical presentation and the easy diagnosis if suspected. PMID- 10459434 TI - Late complications. Postpneumonectomy syndrome. AB - Postpneumonectomy syndrome is an unusual complication of pneumonectomy characterized by excessive mediastinal displacement towards the empty pleural space with secondary tracheobronchial compression. Successful management involves repositioning of the mediastinum through the use of an intrapleural prosthesis. PMID- 10459435 TI - Late results. AB - Pneumonectomy is performed for a number of benign and malignant conditions. It is most commonly performed for lung cancer. Adjuvant and neoadjuvant protocols have increased the number of these operations being performed and the long-term results are improving. Pneumonectomy may also be performed for metastases to lung and for mesothelioma with encouraging results. Some bronchial adenomas require pneumonectomy. Treatment of resistant mycobacteria or the complications of tuberculosis frequently require pneumonectomy. Late bronchopleural fistulae, esophagopleural fistulae, and empyema may occur. PMID- 10459436 TI - Does gender discrimination exist in a gynecology training program in a private hospital? AB - PURPOSE: Does gender discrimination by attending physicians exists in a residency in regard to residents' opportunities to perform complete/operative management of hysterectomies versus just being surgical assistants? MATERIALS AND METHODS: The program studied is a 4-year program in obstetrics and gynecology residency with 3 residents per year. All cases involving a resident were recorded in a computer program designed by one of the authors (C.S.M.) to collect data for Residency Review Committee reports. Data were able to be sorted in a variety of methods including level of management, date of procedure, Physicians' Current Procedural Terminology codes, and attending physician name or resident name. Only intrafascial and extrafascial hysterectomies for benign disease were included in the study. Data were collected from July 1, 1996 to March 31, 1997. RESULTS: Five hundred and forty-nine hysterectomies with residents participating as primary surgeon (complete/operative management) or surgical assistant were performed during the study period. Complete/operative management was performed by the resident in 82.5% of cases while the resident was surgical assistant in 17.5%. Male residents were responsible for complete/operative management in 81.6% of cases and female residents in 83.2% of cases (P = 0.33). Male attending physicians were more likely to allow residents (male or female) to participate as the primary surgeon in abdominal hysterectomies (95.3%) and vaginal hysterectomies (68.5%) than female attending physicians (abdominal, 87.0% and vaginal, 57.3%) (P < 0.001 and P = 0.006, respectively). CONCLUSIONS: Although male attending physicians were more likely than female attending physicians to allow residents to perform complete/operative management, there was no discrimination as to whether the resident in question was male or female. PRECIS: When determining the level of management private gynecologists will allow residents to perform they do not practice gender discrimination. PMID- 10459437 TI - Immunohistochemical detection of apoptosis in the human term and post-term placenta. AB - PURPOSE: To study the incidence of apoptosis in human term and post-term placenta and to determine its presence in different areas of placentas of uncomplicated pregnancies. METHODS: A total of 15 placentas, 8 obtained from spontaneous deliveries and elective caesarean sections at term (37-41 weeks of pregnancy) and 7 from spontaneous deliveries and elective caesarean sections post-term (> 41 weeks of pregnancy) were included in this study. Apoptosis was identified by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method (TUNEL, Boehringer, Mannheim, Germany) in paraffin-embedded sections. RESULTS: Apoptosis was predominantly detected in trophoblast and stromal tissue. There were no significant differences in the incidence of apoptosis in different parts of placental tissue. A significant increase of apoptosis was seen in both trophoblast and stromal cells of post-term placentas (p < 0.05; p < 0.005). CONCLUSIONS: Apoptosis could be detected in the human term and post-term placenta, with increasing incidence in post-term placental tissue, suggesting a possible role of apoptosis in the mechanism of parturition and placental senescence. PMID- 10459438 TI - Colposcopy, cytology and histology in the diagnosis of squamous intraepithelial lesions of the cervix. AB - OBJECTIVE: To compare colposcopic findings to cytologic and histological diagnoses in women with colposcopic reports of ANTZ and/or HPV infection. METHODS: Among 791 hospitalized women referred for colposcopic examination, colposcopy showed ANTZ grade 0-2 and/or HPV infection in 271 patients (34.26%). Only 153 were fully investigated by colposcopy, cytology (under colposcopic observation) and histology (target punch biopsy: 109 patients; surgical specimens of hysterectomy: 42 patients; conization: 2 patients). Cytological and histological diagnoses were reported according to the Bethesda System. RESULTS: 132/153 Pap smears were estimable for sampling adequacy; 44/63 resulted as normal and were histologically positive for LSIL [1]. Five LSIL-positive Pap tests were negative on histology (false negative and false positive rate of 33.33% and 3.78%). The pap test was diagnostic for intraepithelial neoplasia in 34/65 cases (53.3%) and for invasive cancer in 6/11 cases (54.5%). In 67/132 cases (50.8%) adequate-for-sampling Pap smears could not predict the exact diagnosis. On the other hand, 108/141 patients with colposcopic evidence of ANTZ/cancer showed histological SIL or invasive neoplasm (76.59%): ANTZ 1 was associated to LSIL and HSIL in 74.1% and 2.4%; ANTZ 2 to LSIL, HSIL and invasive cancer in 41%, 30.76% and 10.3%. Colposcopic suspicion of invasive cancer in 8 patients was histologically demonstrated in 7 (87.5%); colposcopic diagnosis of HPV infection was confirmed in 10/12 (83.4%). CONCLUSION: A better correspondance was shown between colposcopy and histology than between cytology and histology in the diagnosis of SIL. We suggest a routine colposcopy investigation for all patients admitted to a gynecological clinic and we believe it is very important to take Pap smears under colposcopic observation if colposcopy and cervical smears are performed in the same sitting. PMID- 10459439 TI - Danazol effects on human endometrial cells in vitro. AB - Although danazol has been reported to inhibit endometrial cell growth in vitro, it is difficult to accept that this is through direct inhibition of danazol on endometrial cells. This is because local danazol therapy improves endometriotic signs and symptoms without any disturbance in ovulation or the menstrual cycle. We have re-evaluated the effects of danazol on human endometrial cells by using normal human endometrial stromal cells and two cell lines derived from highly differentiated endometrial adenocarcinomas. Danazol is difficult to dissolve in aqueous solutions, and undissolved danazol significantly inhibited endometrial cell growth even at less than 100 ng/ml. At 500 ng/ml of fully dissolved danazol, which is the therapeutic dose, danazol solution inhibited leukemic cell growth but not endometrial cell growth. Therefore, the action of danazol may be not an inhibitory effect but rather a regulatory function of endometrial cell growth. PMID- 10459440 TI - Evaluation of stress-related hormones after surgery. AB - INTRODUCTION: Improvements in instrumentation, optics, video cameras and technology have brought laparoscopy to the point at which many surgical procedures that once could be performed only by laparotomy are now successfully performed endoscopically. PATIENTS AND METHOD: Twenty women undergoing surgery by laparotomy (11 patients) or laparoscopy (9 patients) were evaluated prospectively. Concentrations of adrenaline, noradrenaline, cortisol, prolactin, and ACTH were measured. Statistical analysis was performed by using the Friedman Two-way ANOVA, Kruskal-Wallis 1-way ANOVA and Mann-Whitney U-Wilcoxon Rank Sum W Test. RESULTS: By comparing the means of all serum levels of ACTH, cortisol and prolactin between the laparoscopy and the laparotomy group, significant differences of ACTH (p < 0.0001), prolactin (p = 0.0164) and cortisol (p < 0.0001) were found. Furthermore, statistically significant differences (p < 0.05) of the serum levels of noradrenaline and adrenaline between the laparoscopy and the laparotomy group were observed. DISCUSSION: While laparoscopic surgery causes less activation of stress-related hormones laparotomy results in a much more obvious response to all hormones evaluated, particularly catecholamine and ACTH production. This is probably related to the major tissue trauma. PMID- 10459441 TI - Laparoscopic evaluation and management of chronic pelvic pain during adolescence. AB - Chronic pelvic pain (CPP) is a common symptom and a difficult condition to manage especially during adolescence. The aim of this study was to evaluate the role of laparoscopy in the diagnosis and treatment of CPP during this period of life. From January 1993 to December 1997, 98 patients, selected from a group of 180 patients who were referred to our clinic underwent laparoscopy. In most cases (60%) no abnormalities were observed. Endometriosis was found in 25% of cases, followed by ovarian cysts 7%, parovarian cysts 3%, pelvic inflammatory disease 3% and adhesions 2%. Laparoscopic treatment was performed as indicated by laparoscopic findings. We conclude that laparoscopy is a valuable and effective procedure in the diagnosis and management of CPP in a selected group of patients. PMID- 10459442 TI - Pelvic abnormalities in hysterectomized patients-role of early postoperative ultrasonographic evaluation. AB - OBJECTIVE: To assess the incidence of post-hysterectomy pelvic fluid collection in the early postoperative period and to study its natural history. STUDY DESIGN: Cross-sectional, prospective, observational study. PATIENTS AND METHODS: The study sample comprised 36 consecutive patients undergoing hysterectomy for benign conditions in our department. All patients underwent two pelvic ultrasonographic examinations, the first on the third postoperative day and the second one year later. All postoperative complications were thoroughly evaluated. RESULTS: Thirty two patients completed the study evaluations. Abnormal ultrasonographic findings were detected in 4 of them on the early postoperative scan. Three (9.4% of the sample) had pelvic fluid collections which persisted on the follow-up scan. The fourth patient had a simple ovarian cyst 4 cm in diameter which disappeared on follow-up. None of the patients with pelvic fluid collections had a febrile morbidity during the postoperative course. CONCLUSIONS: Transvaginal ultrasonography can detect asymptomatic early postoperative pelvic fluid collections and enables conservative management, thereby reducing patients stress, medical costs, and the need for unnecessary interventions. PMID- 10459443 TI - Pain-relief and movement improvement by acupuncture after ablation and axillary lymphadenectomy in patients with mammary cancer. AB - INTRODUCTION: In the traditional Chinese medicine (TCM), pain and movement restrictions are considered as the result of a blocking of the "Jing-Luo-system" or of a disordered flow of the Jing-Qis in the "Jing-Luo-system". PATIENTS AND METHODOLOGY: In this study 48 patients with mammary cancer after ablation and axillary lymphadenectomy were treated with acupuncture (group I); a control group of 32 patients with the same operation but without acupuncture was compared (group II). RESULTS: The results showed a significantly higher maximum abduction angle (AA) at the first treatment immediately after acupuncture without pain (59.1 degrees vs. 80.4 degrees, p < 0.001) with respect to maximum tolerable pain barrier (73.6 degrees vs. 92.3 degrees, p < 0.001). Between group I (12.3%) and group II (50%) there was a statistically significant difference (p < 0.01) in the appearance of pain in the operation field in the rest position on the 5th postoperative day, while on the 7th postoperative day 8.3% vs. 12.5%) and at the time of discharge a significant difference could not be seen (p > 0.05). The percentage of patients with pain during arm movements showed a statistically significant difference between group I and group II on the 5th postoperative day (81% vs. 100%, p < 0.01), on the 7th postoperative day (43% vs. 96.9%, p < 0.01) and at time of discharge (27.1% vs. 65.6%, p < 0.001). The differences in the abduction angle between group I and group II were also statistically significant on the 5th postoperative day at indolency (89.3 degrees vs. 74.5 degrees, p < 0.001) with respect to maximum tolerable pain (105.8 degrees vs. 87.4 degrees, p < 0.001). The differences in the abduction angle on the 7th postoperative day at indolency (97.5 degrees vs. 81.2 degrees, p < 0.001) and at maximum tolerable pain (118.5 degrees vs. 93.4 degrees, p < 0.001) were statistically significant. This statistically significant difference in the maximum abduction angle between group I and group II at indolency (116.1 degrees vs. 91.5 degrees) with respect to maximum tolerable pain (129.4 degrees vs. 112.7 degrees, p < 0.001) could be observed until discharge. DISCUSSION: Acupuncture seems to be an effective treatment to relieve pain and improve arm-movements after ablation and axillary lymphadenectomy. The "Xie-technique" is used at the main acupuncture points and the patient's feeling must be particularly considered. The combination of the different main points with the correctly selected additional acupuncture points- referred to the basic state and the pre- und post-operative state of the patient- are very important for a successful application of acupuncture. PMID- 10459444 TI - Evaluation of combined endoscopic and pharmaceutical management of endometriosis during adolescence. AB - Laparoscopy is the most frequent surgical approach in gynecologic patients with acute or chronic pelvic pain. The symptomatology is frequently related to a specific gynecological pathology such as endometriosis or associated adhesive disease. During an eight year period, January 1990 to December 1997, 26 patients (aged 16-20 years) with endometriosis were diagnosed endoscopically and managed pharmaceutically in our clinic. The disease was evaluated and staged according to the American Society of Reproductive Medicine. The disease was evaluated as first stage in 16 patients (61.6%), as second stage in eight patients (30.8%), as third stage in one patient (3.8%) and as fourth stage in one patient (3.8%). Patients underwent adhesiolysis and management according to their laparoscopic findings. Postoperative pharmaceutical treatment (Danazol, GnRH analogues, Oral Contraceptives) was given. Patients were followed for the evaluation of the treatment. The efficacy of the combination of endoscopic and pharmaceutical management of the disease is discussed. PMID- 10459445 TI - Postmenopausal users of long-term hormonal replacement therapy: social-cultural features. AB - OBJECTIVE: To study of influence of social-cultural factors on the effects of long-term hormonal replacement therapy (HRT) in preventing cardiovascular disease and menopausal osteoporosis. METHOD: We examined, perspectively the social cultural features of 394 postmenopausal women who attended the menopausal out patient department and were eligible for long term HRT. RESULTS: The HRT users were thinner and younger, with earlier menopause, physically more active, healthier, with more oophorectomies and with more professional work. Data shows that, as a whole, a greater number of healthy women are selected or self-selected for HRT. CONCLUSION: Evaluating the results of long-term HRT in preventing cardiovascular disease and menopausal osteoporosis, the social-cultural features of HRT users must be carefully considered before any mass preventive interventions. PMID- 10459446 TI - Day clinic diagnostic hysteroscopy in a state hospital. AB - Within 6 years (1991-97), a total of 680 diagnostic hysteroscopies were performed at the Day Clinic of the 1st Department of Obstetrics and Gynecology of the University of Athens ("Alexandra" Hospital). The procedure was done without general or other forms of anesthesia, using the Siegler method of approach. However in 12 cases with cervical stenosis (1.7%) and 21 patients with marked nervousness (3.1%) general anesthesia proved inevitable. The leading indication was repeated failure of IVF (54.7%), while other indications included abnormal bleeding, amenorrhea and oligomenorrhea, a history of abortions, and infertility. Abnormal hysteroscopic findings were observed in 276 cases (40.5%) among which intrauterine adhesions, endometrial hyperplasia and polyps were the most common. We had no major complications or fatalities in our series and hysteroscopy proved to be a very useful, accurate and safe method of assessing uterine and endometrial functional status. PMID- 10459447 TI - Ovarian stimulation using a new highly purified urinary FSH: a prospective randomized clinical study. AB - The aim of this study was to determine the effectiveness of a new highly purified urinary FSH. A total of 60 in vitro-fertilization (IVF) patients, undergoing embryo transfer (ET) for the first time, were randomly allocated into two groups: Group A (n = 30). Subcutaneous administration of urinary follicle-stimulating hormone (FSH, Fostimon 75, A.M.S.A., Italy). Group B (n = 30). Subcutaneous administration of urinary follicle-stimulating hormone (FSH, Metrodin 75 HP, Serono, Italy). Statistical analysis was performed using the chi-square test, p < 0.05 was assumed as significant. This prospective randomized clinical study in an IVF-ET program showed that both drugs were equally safe and effective. Except for the number of the high quality embryos (3.16 vs 2.9; p = 0.03) the two groups did not differ in stimulation parameters or clinical pregnancy rates per attempt and per transfer. On the other hand, a mean number of 3.56 vs 2.18 embryos were cryopreserved in group A and in group B, respectively, as a result of the high number of mature oocytes and high quality embryos. When frozen embryos cycles were included, the difference in pregnancy rate became significant. PMID- 10459448 TI - Gestational diabetes mellitus: intensive versus mild treatment. PMID- 10459449 TI - The role of intravenous albumin in the prevention of severe ovarian hyperstimulation syndrome. A pilot experimental study. AB - OBJECTIVE: To evaluate the role of intravenous albumin in the prevention of severe ovarian hyperstimulation syndrome (OHSS). DESIGN: A pilot experimental study. MATERIAL AND METHODS: Ovarian hyperstimulation was induced in 5 rabbits using human menopausal gonadotropin/human chorionic gonadotropin, after pretreatment without (control group) or with bovine serum albumin (BSA group). RESULTS: Despite an increase in serum protein levels, the BSA group showed comparable delta increase in body weight and degree of ascites formation. CONCLUSIONS: Intravenous albumin did not prevent severe OHSS in a rabbit model despite the observed increase in serum oncotic pressure. PMID- 10459450 TI - Does maternal drug ingestion cause megacystis microcolon intestinal hypoperistalsis syndrome? I. Clomiphene trial. AB - PURPOSE: Megacystis Microcolon Intestinal Hypoperistalsis Syndrome (MMIHS) is a congenital and lethal disease, and the etiology of the disease is not clear. It is speculated that maternal ingestion of some drugs during pregnancy may be an etiologic factor. In this study we aimed to investigate the effect of maternal ingestion of clomiphene on fetal bladder and colon in pregnant rats. METHODS: We separated animals into a control group including 14 rats and a clomiphene group with 30 rats. Nothing was given to the control group during pregnancy. Two mg/kg/day clomiphene intraperitoneally was given to the study group from the 6th to 12th day of pregnancy. All of them were sacrificed on the 20th day of pregnancy. Histopathological examination of the fetal colon and bladder was performed. RESULTS: In the clomiphene group a significant decrease in the thickness of the bladder wall, an increase in bladder epithelium, an increase in muscle atrophy of the colon and bladder wall, an increase in vacuoler degeneration of the muscles of the bladder and colon wall, a decrease in ganglion cell numbers in the myenteric plexus of the bladder and a decrease in the thickness of the bladder tunica muscularis were determined. CONCLUSION: In our rat model we found histological structural changes in the rats' colon and bladder walls as a result of using clomiphene on days 6-12 of pregnancy; a similar pathological finding to those found in some of the MMIHS patients' colons and bladders. PMID- 10459451 TI - Breast cancer and replacement therapy: which women are at risk? AB - The fear of breast cancer is the most important concern for women who have to decide whether to take hormone replacement therapy (HRT) for the treatment and prevention of postmenopausal disturbances. A calculation of the risk for breast cancer with respect to HRT is useful to reassure women about this risk and to help them to decide. In the present report, all breast cancer risk factors have been examined and those likely worsened by increased levels of estrogens with HRT have been considered. On the basis of pathogenic, clinical and epidemiological evidence seven breast risk factors (testosterone levels, body mass index, waist to-hip ratio, alcohol consumption, density to mammography, previous benign breast disease, familiarity) have been selected and a score has been awarded to them; then a model for the calculation of the SRR (Summary Relative Risk) has been elaborated. A simple, feasible, easy to achieve decision model for the calculation of breast cancer risk is proposed in relation to the use of HRT. PMID- 10459452 TI - Growth hormone response to oral glucose tolerance and insulin tolerance tests in patients with PCOS. AB - Polycystic ovary syndrome (PCOS) is characterized by its heterogeneity. This is the reason for the diversity in the clinical manifestations and laboratory findings. In this study we examined the serum levels of growth hormone (GH) in 15 women with PCOS and 5 healthy volunteers following oral administration of 75 gr glucose (OGTT) and intravenous administration of insulin (ITT). The OGTT produced no significant difference between the two groups of women, in the ITT there was a difference between the GH response of the women with PCOS and that of the healthy women. The group with PCOS showed a later, more prolonged, higher response, indicating that the hypothalamus is probably involved in PCOS. PMID- 10459453 TI - Strumal carcinoid of ovary. A case report with immunohistochemical investigations. AB - Clinical, histological and immunohistochemical findings in a rare ovarian tumor- strumal carcinoid--are presented. Neuropeptides detected by immunoreactions revealed the origin of tumor cells from the midgut. Immunohistochemical reaction revealed parathyroid hormone-related protein (PTHrp) in the carcinoid cells. After surgical resection of the tumor a decrease in serum calcium level was observed. PMID- 10459454 TI - Treatment results of liquid nitrogen cryotherapy on selected pathologic changes of the uterine cervix. AB - One hundred and eighty-two women treated with cryotherapy were included in the study. In cases of chronic cervical inflammation full recovery was observed in 86% of subjects. In cases of Naboth cysts full recovery was observed within 6 to 8 weeks in 89% of women. In cases of cervical erosions full recovery was observed within 6 to 9 weeks in 84.9% of women. In women, with cervical epithelium dysplasia full recovery was observed within 7 to 11 weeks in treated subjects. Renewal of the pathology was observed in 15% of the women with treated cervicitis and in one of the women with cervical dysplasia within 2 to 5 years of follow-up. PMID- 10459455 TI - Congenital absence of breast and nipple in a woman 32 years old. PMID- 10459456 TI - Serum soluble CD23 and TNF-alpha in women with spontaneous abortion in the first trimester. AB - The present study was designed to investigate the participation of sCD23 and TNF alpha in pregnant women in the first trimester. One hundred and forty-one patients were enrolled in the study. Women were classified into two groups. Serum sCD23 and TNF-alpha were measured in 73 normal pregnant women in the first trimester (Group I) and in 68 women with spontaneous abortion (Group II). We found that the mean values of TNF-alpha levels were higher in women with normal pregnancy than in women with spontaneous abortion (p < 0.05), whereas sCD23 levels of women in group II did not differ statistically from the controls. These results sustain the opinion that women with spontaneous abortion seem to have reduced TNF-alpha levels. However, determination of these immunological parameters provides no useful clinical information about disease activity and management. PMID- 10459457 TI - Colposcopic diagnosis: clinical aspects and experiences. AB - Colposcopy can be applied as an integral part of every gynecologic examination in concert with cytology; to identify and localize lesions suspected on the basis of abnormal cytology findings; and to clarify the nature of clinically suspicious lesions. This implies using colposcopy to clarify the nature of cervical changes seen with the naked eye. This practice is superior to colposcopy which is restricted to evaluating abnormal smears because it can pick up some lesions missed by cytology. But it is not as effective as routine colposcopy because it can miss lesions not picked up by gross inspection of the cervix and because there is no opportunity to inspect the lower tract of the cervical canal. Considering that only 15-20% of lesions are purely endocervical, not too much time is wasted by examining these cases. If colposcopy is limited to evaluating grossly suspicious lesions, then its role is merely to avoid unnecessary biopsies. PMID- 10459458 TI - Herpes simplex genitalis type 2: our experiences. AB - OBJECTIVE: To review the recent research in genital HSV infection and to pay attention to this problem in Slovene population. METHODS: Review of all papers in the field on Medline 1994-98. A descriptive retrospective analysis of symptoms, seroprevalence, treatment, complications and follow-up of 49 HSV-1 and HSV-2 seropositive patients. RESULTS: Five patients with overt genital herpes were HSV 1 seropositive, and four were HSV-1 and HSV-2 seropositive. One patient had extragenital spread and HSV proctitis. Treatment consisted of 1 gr daily dose of Acyclovir (ACV) in combination with topical ACV for two to three weeks. Recurrences were intermittently treated in three patients and one started suppressive treatment. Three pregnancies terminated vaginally with healthy babies. CONCLUSIONS: According to literature overview and our experiences, more attention should be paid on revealing mild forms of genital herpes in order to diminish the spread of infection and its consequences. PMID- 10459459 TI - Treatment of vaginal intraepithelial neoplasia (VAIN) with the carbon dioxide laser. AB - The increasing incidence of VAIN especially in young women, the frequent relapses, and renewed interest in maintaining sexual function have prompted gynecologists to a conservative management of the disease. Over the last decades, surgery, 5-fluorouracil, chemosurgery, electrocautery and cryotherapy were used. Carbon dioxide laser ablation therapy of VAIN has been reported from various authors with different results. From June 1991 through December 1998, 39 patients affected by VAIN were treated with laser surgery (35 vaporizations and 4 excisions). To achieve complete elimination of all lesions, seven patients had two vaporizations and one patient three. One patient was submitted to six combined repeated treatments. Five patients were not evaluable and three presented persistence of VAIN. One patient died because of AIDS. The remaining 30 patients, treated with laser surgery, were lesion free: 7 patients were negative at 12-24 months, 10 at 24-36 months and 13 at 37-90 months. No important complications occurred. Sexual function was not compromised. Carbon dioxide laser is a safe and efficacious tool in the treatment of pre-neoplastic lesions of the vagina. PMID- 10459460 TI - Sexually-transmitted diseases and assisted reproduction techniques. AB - As to the correlation between sexually transmitted diseases (STD) and assisted reproduction techniques (ART) we must consider that it is necessary to collate information and start screening campaigns for all sexually active individuals with the purpose of prevention and early diagnosis of STD which may cause of sterility. Before starting any kind of ART it is mandatory to perform diagnostic tests in all couples and to apply the correct therapy for any STD: treatment must always involve both partners and results must be carefully checked. All diagnostic examinations and all ART procedures should not carry the risk of transmitting any infection. When one or both of the partners are carriers of a transmissible infection (especially AIDS) it is advisable to negate ART or to start treatment, particularly for the semen, in order to reduce the level of infection and to avoid the transmission of any infection to the fetus. PMID- 10459461 TI - Epidemiology of bladder control problems. PMID- 10459462 TI - Stress urinary incontinence. AB - New techniques for the treatment of female stress urinary incontinence are constantly being developed. In assessing treatment modalities, one must consider many variables including patient satisfaction and quality of life, cure or improvement rate, patient selection, and long-term durability of results. In 1998, many papers comparing and examining the outcomes of various new and old techniques were published. PMID- 10459463 TI - Overactive detrusor. AB - Detrusor overactivity poses a major challenge to physicians from many specialities. Progress in our understanding of the pathogenesis of detrusor overactivity is slow but steady. Advances in treatment continue to be made, both with new agents and new methods of delivering older drugs. Neuromodulation is showing great promise as experience increases, and developments in bladder augmentation offer hope for lower morbidity treatments in future. PMID- 10459464 TI - Interstitial cystitis. AB - Interstitial cystitis is a complex inflammatory condition of the bladder. The pathophysiology of interstitial cystitis is incompletely understood, although altered epithelial permeability, mast cell activation and sensory afferent nerve upregulation play critical roles. A unified understanding of the pathogenesis of interstitial cystitis is emerging and this will hopefully lead to the introduction of novel therapies for pain and irritative voiding symptoms. Interstitial cystitis is a common disease among women and is frequently misdiagnosed as prostatitis and benign prostatic hyperplasia among men. PMID- 10459465 TI - Neurogenic bladder dysfunction. AB - Urodynamics are still essential for diagnosis and prognosis of neurogenic lower urinary tract-dysfunction and can not be replaced by other means of investigation so far, neither by a thorough clinical investigation nor by sophisticated means like magnetic resonance imaging. The findings with clinical investigations are specific, but not sensitive enough, and the spinal cord lesions may sometimes be beyond the resolution of magnetic resonance-scanning. Pharmacotherapy is still the corner stone in the management of detrusor hyperreflexia. Further studies with tolterodine, oxybutynin, trospiumchloride and propiverine have increased our knowledge about these substances. Capsaicin was proved to be the effective substance and not the alcoholic solution, which serves as a carrier. Intrathecal clonidine may represent a new conservative reversible alternative treatment for detrusor hyperreflexia. Experiments with detrusor strips from end-stage MMC patients may explain the relative resistance of the low compliant bladder to the common anticholinergic/spasmolytic therapy. The differential indication for bladder augmentation, either using segments of the gastrointestinal-tract or performing a partial detrusor myectomy is ongoing, favourable results are reported for both techniques. Sacral posterior root rhizotomy is able to abolish detrusor hyperreflexia and therefore recommended for tetra- and paraplegics, however autonomic dysreflexia, if present, can not be totally abolished. Collagen injections for neuropathic sphincter incompetence can not be recommended as demonstrated in children with congenital neuropathy, a new design of an artificial sphincter must stand the test of time. PMID- 10459466 TI - Orthotopic bladder augmentation and substitution. AB - Orthotopic bladder augmentation or substitution using intestinal segments has become a standard procedure for many disorders that cause a loss of functional or anatomical bladder capacity. From the technical point of view, reservoir configuration by detubularizing the intestinal segments is the general practice. Various techniques exist, depending which types of segments and which techniques of ureteral implantation are used. Common problems include urinary incontinence, retention, metabolic disorders, and the possibility of secondary malignancies. As a result, research has been conducted into utilizing tissues other than intestine for bladder augmentation or substitution. PMID- 10459467 TI - Urinary stones. PMID- 10459468 TI - Current advances in shock wave lithotripsy. AB - The introduction of shock wave lithotripsy into clinical practice revolutionized the management of urinary tract stone disease. As experience has been gained with its use, however, the limitations and adverse effects associated with it have been recognized. PMID- 10459469 TI - Percutaneous nephrolithotripsy. AB - Percutaneous nephrolithotripsy is a well established technique for the treatment of the stones resistant to ESWL. Simultaneous bilateral procedures, supracostal approach and the 'mini-perc' technique are interesting technical improvements. Percutaneous nephrolithotripsy could in future be the primary treatment not only of large burden stones, but also for lower caliceal stones larger than 1 cm. PMID- 10459470 TI - Ureteroscopic lithotripsy. AB - The indications for ureteroscopic lithotripsy have increased with endoscope miniaturization and powerful, precise endoscopic lithotrites like the holmium: yttrium-aluminum-garnet laser. Successful ureteropyeloscopic treatment with the currently available instrumentation and techniques is independent of the size, composition, and location of stones in the upper urinary tract. Extracorporeal shockwave lithotripsy maintains a major role in treating uncomplicated, moderately sized upper urinary tract calculi. Complex upper urinary tract calculi, however, are best treated endoscopically. In addition, the endoscopic treatment of ureteral calculi is efficacious and definitive, albeit more invasive than extracorporeal shock wave lithotripsy. PMID- 10459472 TI - Bibliography. Current world literature. Voiding dysfunction and female urology. PMID- 10459471 TI - Metabolic evaluation and medical therapy for stone formation. AB - There is little debate that nephrolithiasis is an increasing problem for public health. In the USA, the treatment of renal stone disease costs more than two billion dollars per year. The lack of prospective randomized trials reflects the problem of subjecting stone-formers to prospective studies and placebo treatment, and studies often use a single-group design. Nevertheless, investigation of metabolic abnormalities often makes the identification of an etiologic factor, and consequent treatment, possible, and stone formation can be reduced. PMID- 10459473 TI - Bibliography. Current world literature. Stones. PMID- 10459474 TI - Sudden cardiac death. AB - SCD continues to be an important cause of death and morbidity. Despite expanding insight into the mechanisms causing SCD, the population at high risk is not being effectively identified. Although there is still much to do in the management phase of SCD (predicting the efficacy of various therapies), recent clinical trials have helped define the relative risks and benefits of therapies in preventing SCD. Trials are underway to determine whether treating other patient populations, including asymptomatic patients after MI, will improve survival rate. The approach to reducing mortality rate will always be multifaceted; primary prevention of coronary artery disease and prompt salvage of jeopardized myocardium are 2 important aspects of this approach. In addition to interventions for MI, such as myocardial revascularization when indicated, simple and easily administered therapies that are likely to remain the most effective prophylactic interventions are aspirin, ACE inhibitors, beta-blockers, and cholesterol lowering agents. However, the MADIT and AVID data clearly demonstrate a role for ICD therapy in a subgroup of patients who have VT/VF and are at risk of cardiac arrest. Even though the absolute magnitude of benefit associated with ICDs is still to be determined, the AVID study and other recent reports provide convincing evidence that patients who have VT/VF fare better with ICDs than with antiarrhythmic drug therapy. For the high-risk population described in this article, in addition to aggressive anti-ischemic and heart failure therapy, ICDs are now a mainstay of life-saving treatment. Still to be surmounted is the challenge of identifying patients who have nonischemic substrates and of providing them with the appropriate therapy. Guided by genetic studies and new insight into the mechanisms of such problems as congenital long QT syndrome, life saving and life-enhancing therapies may soon be available for the management of SCD. PMID- 10459475 TI - Comparative evaluation of an automated ribotyping instrument versus pulsed-field gel electrophoresis for epidemiological investigation of clinical isolates of bacteria. AB - A collection of bacterial isolates were typed using the Ribo-Printer Microbial Characterization System (Qualicon, Wilmington, DE, USA), an automated ribotyping system, and pulsed-field gel electrophoresis (PFGE). Grouping patterns, discrimination, and typeability were compared. The collection consisted of 411 isolates of bacteria from 32 medical centers. The isolates included a total of 18 species (both Gram-positive and Gram-negative), covering the range of concern to a laboratory performing epidemiological investigations. The patterns of groups obtained by both typing systems were similar for all species examined. Ribotyping provided less discrimination than PFGE, especially for Serratia marcescens, and Pseudomonas aeruginosa. All strains were typed by the Ribo-Printer, but 2.75% were not typeable by PFGE. The Ribo-Printer has proven to be a valuable primary typing method for a high-volume laboratory, even for those species for which it provides a lesser degree of discrimination than PFGE. PMID- 10459476 TI - Phenotypic characteristics of Enterococcus faecium variants confirmed by intergenic ribosomal polymerase chain reaction and E. faecium polymerase chain reaction. AB - Enterococcus faecium has recently emerged as a serious nosocomial pathogen. The emergence of multiple antimicrobial agent-resistant E. faecium has been remarkable; with its strains it is one of the most phenotypically heterogeneous of all enterococcal species. About 15% of enterococcal strains isolated from human clinical specimens were found to have atypical biochemical characteristics. In order to determine if these strains were E. faecium variants, intergenic ribosomal polymerase chain reaction (ITS-PCR) and E. faecium PCR (EfPCR) were performed in 45 atypical strains, and the two PCR results were used to analyze phenotypic characteristics of the strains. As many as 60% (27/45) of the atypical strains were identified as E. faecium. Thus, it is concluded that if an enterococcal strain shows positive reaction to arabinose, arginine, and ribose and negative reaction to methyl-alpha-D-glucopyranoside and pigment, it should be identified as E. faecium. PMID- 10459477 TI - Evaluation of the Strep A OIA assay versus culture methods: ability to detect different quantities of group A Streptococcus. AB - The Strep A OIA assay by Biostar (Boulder, Co., USA) is a unique optical immunoassay system for the rapid detection of Group A streptococcal carbohydrate. As part of a community-based pediatric cohort study of Group A Streptococcus (GAS) persistence following antibiotic therapy of pharyngitis, the performance of the Strep A OIA assay was compared with the amount of growth from standard throat swab culture methods. A total of 363 throat swabs taken over the course of the study was evaluated from 248 children between 2 and 18 years of age. Two culture methods were performed: an agar plate with the throat swab using Columbia agar base with 5% sheep blood incubated under an anaerobic environment for 48 h and Todd-Hewitt broth (THB) enhancement. The Strep A OIA was then performed. A total of 144 of 363 (39.7%) samples was positive for GAS by one or more of the laboratory tests across study visits: agar culture detected 132 of 144 (91.7%), THB culture detected 128 of 144 (88.9%), and the Strep A OIA assay detected 129 of 144 (89.6%). Complete agreement among all three laboratory tests was found for 333 of 363 (91.7%) of the samples. Agar culture results were comparable to THB cultures with a sensitivity of 96.9%, specificity of 96.6%, a positive predictive value of 93.9%, and a negative predictive value of 98.3%. Although the performance of the Strep A OIA assay had similar specificity (96.5%) and positive predictive value (93.8%) compared with the combined results of the two culture methods, the sensitivity (89.0%) and negative predictive value (93.6%) were lower. A significant difference (p < 0.001) was found in the ability of the Strep A OIA assay to detect agar culture-positive swabs that had a light growth (1+ or 2+) (63.0%) versus a moderate (3+) or heavy (4+) growth (98.1%) of GAS. Although the Strep A OIA assay allows GAS throat swab results to be reported an average of 24 h sooner than either of the cultures, the rapid assay was not as sensitive in detecting light growth GAS-positive cultures. PMID- 10459478 TI - High rate of non-albicans candidemia in Brazilian tertiary care hospitals. AB - In order to evaluate the epidemiology of candidemia in Brazil, we performed a prospective multicenter study conducted in six general hospitals from Sao Paulo and Rio de Janeiro, We enrolled a total of 145 candidemic patients (85 males) with a median age of 32 years. Non-albicans species accounted for 63% of all episodes and the species most frequently causing candidemia were C. albicans (37%), C. parapsilosis (25%), C. tropicalis (24%), C. rugosa (5%), and C. glabrata (4%). Systemic azoles were used before the onset of candidemia in only six patients. There were no differences in the coexisting exposures or underlying diseases associated with the species most frequently causing candidemia. The overall crude mortality rate was 50%. Nosocomial candidemias in our tertiary hospitals are caused predominantly by non-albicans species, which are rarely fluconazole resistant. This predominance of non-albicans species could not be related to the previous use of azoles. PMID- 10459479 TI - Use of reverse transcription polymerase chain reaction with colorimetric plate hybridization to detect a cytomegalovirus late spliced mRNA in polymorphonuclear leukocytes from renal transplant patients. AB - Human cytomegalovirus replication was evaluated in polymorphonuclear leukocytes from ten renal transplant recipients. Three new reverse transcription polymerase chain reactions with plate hybridization suitable for automation were developed for the detection of immediate-early spliced UL123 mRNA, early-late pp65 mRNA, and late spliced UL22 mRNA. The presence of UL22mRNA was found to be significantly associated with the occurrence of cytomegalovirus (CMV) disease. PMID- 10459480 TI - Multicenter evaluation of the MB-Redox medium compared with radiometric BACTEC system, mycobacteria growth indicator tube (MGIT), and Lowenstein-Jensen medium for detection and recovery of acid-fast bacilli. AB - MB-Redox is a new manual culture system designed for the recovery of mycobacteria from clinical specimens. It consists of a liquid medium (modified Kirchner medium) containing a redox indicator, a colorless tetrazolium salt, which is reduced to colored formazan by actively growing mycobacteria. Acid fast bacilli (AFB) are easily detected in the medium as pink to purple pinhead-sized particles. We report the results of a multicenter study (involving four Italian microbiology laboratories processing 2370 clinical specimens) aiming to evaluate the recovery rates of AFB and time required for their detection by using the MB Redox medium. Two different protocols were set up: in Protocol A (1580 specimens) the performance of MB-Redox was compared with those of the radiometric BACTEC 460 TB system (B460) and Lowenstein-Jensen medium (L-J), whereas in Protocol B (790 specimens) it was compared with those of the Mycobacteria Growth Indicator Tube (MGIT) and L-J. A total of 213 mycobacteria were recovered, including 172 Mycobacterium tuberculosis complex (MTB) isolates and 41 nontuberculous mycobacteria (NTM) isolates. In Protocol A, recovery rates were 81% for MB-Redox system, 84% for B460 system, and 77% for L-J. In Protocol B the recovery rates by individual system were 87, 83, and 76% for MB-Redox, MGIT, and L-J, respectively. Differences in both the protocols were not statistically significant. The MB Redox system plus L-J (Combination 1) recovered 94% of the isolates in Protocol A and 93% in Protocol B, while B460 plus L-J (Combination 2) and MGIT plus L-J (Combination 3) detected 91 and 89% of all mycobacteria isolates respectively. No statistically significant differences were found among the combinations. The mean time to detection of mycobacteria was 16.3 days in Protocol A and 19.1 days in Protocol B with the MB-Redox system, 22.4 and 25.9 days with L-J, 13.2 days with B460, and 18.2 days with MGIT. The contamination rates were 2.1, 2.0, 1.9, and 3.6 for MB-Redox, B460, MGIT, and L-J respectively. The MB-Redox is a reliable, nonradiometric system for growth and detection of mycobacteria. When used in combination with a solid medium it proved to be an effective replacement for B460. The MB Redox system is a labor-intensive method requiring much handling during the visual reading procedures. PMID- 10459481 TI - Sitafloxacin (DU-6859a) and trovafloxacin: postantibiotic effect and in vitro interactions with rifampin on methicillin-resistant Staphylococcus aureus. AB - Sitafloxacin (DU-6859a) and trovafloxacin are novel quinolones potent on methicillin-resistant Staphylococcus aureus (MRSA) that are designed for once daily administration. In order to define the adequacy of the above regimen for the therapy of infections by multiple drug-resistant MRSA, their postantibiotic effect (PAE), their bactericidal activity, and their interactions with rifampin were determined on 14 MRSA isolates resistant to both ciprofloxacin and rifampin. PAE was defined after 1-h exposure to 1x, 4x, and 10x MIC and the killing effect after exposure to 1x and 4x MIC. Rifampin was applied for interactive studies at a concentration of 2 micrograms/mL, which is equal to its mean serum level. Median PAEs produced by 1x, 4x, and 10x MIC of sitafloxacin were 1.39, 3.75, and 6.61 h respectively, and by 1x, 4x, and 10x MIC of trovafloxacin 0.87, 2.07, and 2.23 h respectively. PAEs achieved by sitafloxacin were statistically shown to be longer than those achieved by trovafloxacin; PAEs achieved by a concentration of 10x MIC of each quinolone did not differ significantly from those achieved by a concentration of 4x MIC. Both the 4x and 10x MIC concentrations produced a more prolonged PAE than the 1x MIC concentration. A rapid bactericidal activity was expressed over the first 6 h of growth by each quinolone involving 80% of isolates enhanced in some isolates by their interaction with rifampin. The above findings revealed an extended PAE and a rapid killing effect of both sitafloxacin and trovafloxacin on MRSA resistant to ciprofloxacin and to rifampin, thus supporting their once daily administration in the therapy of infections by multiple drug-resistant MRSA. However little in vitro benefit is derived by their interaction with rifampin. PMID- 10459482 TI - Characteristics and trends in macrolide resistance among Helicobacter pylori strains isolated in Bulgaria over four years. AB - Macrolide resistance trends were examined among Helicobacter pylori strains from 154 patients between 1994 and 1998. Applicabilities of screening agar method (SAM) and modified disk diffusion method (MDDM) were evaluated. Overall primary resistance rates to erythromycin and clarithromycin were 14.8% and 8.7%, respectively. No association was found with age, sex, and diseases. Clarithromycin-resistance rate reached 12.5% in the last 2 years. Secondary resistance to erythromycin occurred more often (in 62.5%) than to clarithromycin (in 42.9%). Therapy with spiramycin or erythromycin in four cases induced no clarithromycin resistance. These data show a considerable prevalence of H. pylori resistance to macrolides, which exhibited a tendency to increase and was often associated with metronidazole resistance. By comparing the MDDM with SAM, an overall agreement was obtained in 81 (94.2%) of 86 results. MDDM and SAM are reliable techniques for testing H. pylori susceptibility to macrolides. PMID- 10459483 TI - Antimicrobial activity of gatifloxacin (AM-1155, CG5501), and four other fluoroquinolones tested against 2,284 recent clinical strains of Streptococcus pneumoniae from Europe, Latin America, Canada, and the United States. The SENTRY Antimicrobial Surveillance Group (Americas and Europe). AB - The newer fluoroquinolones generally have greater potency against Gram-positive cocci including Streptococcus pneumoniae. In this study, we report the activity of gatifloxacin (formerly AM-1155 or CG5501) compared with penicillin, erythromycin, and four other peer drugs, tested against 2284 strains isolated in North America (Canada and United States), Latin America (six nations), and Europe in 1997. Reference broth microdilution methods were used and results were interpreted by consensus standards. Gatifloxacin demonstrated uniform potency against pneumococci across all monitored geographic areas (MIC90, 0.5 microgram/mL; > or = 99.6% of strains inhibited at < or = 1 microgram/mL). This activity was comparable to trovafloxacin (MIC90, 0.5 microgram/mL) and sparfloxacin (MIC90, 0.5 microgram/mL) and two- to four-fold greater than that of ciprofloxacin or levofloxacin. The most resistant strains to the fluoroquinolones had mutations in both par C (Ser 79-->Phe) and gyr A (Ser83-->Lys or Phe). Penicillin resistance (MIC, > or = 0.12 microgram/mL) rates varied from 27.6% in Europe to 55.7% in Latin America. Macrolide resistance was greatest in Europe and the United States. Gatifloxacin appears to be a promising new fluoroquinolone for clinical use in respiratory tract infections commonly caused by S. pneumoniae. PMID- 10459484 TI - Pleuro pulmonary infection with Salmonella group E. AB - A patient was admitted with a history of cough, shortness of breath and fever. After investigations, he was found to have a left-sided pneumonia with pleural effusion. Culture of the patient's sputum, pleural fluid and blood revealed Salmonella senftenberg. The patient was started on antibiotics according to the sensitivity report and responded to therapy. The past history revealed attempt at suicide by the intake of corrosive acid, which caused an esophageal stricture. The leak of gastric contents into the mediastinum lead to the infection of the pleural cavity and pneumonia. PMID- 10459485 TI - Cellulitis: evaluation of possible predisposing factors in hospitalized patients. AB - We conducted a prospective study among 62 hospitalized adults, to evaluate the factors that contribute to the development of cellulitis. The majority of patients had multiple possible predisposing factors, and the most common were: diabetes mellitus (31/62), history of cellulitis (30/62), edema (28/62), peripheral vascular disease (25/62), and skin changes suggestive of tinea pedis (20/62). A significant number of patients reported and were clinically noted to have dry skin (42/62). Large controlled studies are needed to evaluate whether aggressive control of possible risk factors can reduce the incidence of cellulitis. PMID- 10459486 TI - Comparison of two selective media for the isolation of Campylobacter species from a pediatric population in Mexico. AB - Two selective media for the isolation of Campylobacter species, a blood containing medium (CampyBAP) and blood-free, charcoal based formulation (CCDA) were compared for the ability to isolate Campylobacter species during a 1-year period. Of the 1,132 stool samples cultured during the study, 42 Campylobacter species were recovered using both media (3.7% yield). CCDA was better than CampyBAP for isolating C. jejuni subsp jejuni (18/20 vs 8/20, P = 0.002) and for all isolates, CCDA was superior over CampyBAP (39/42 vs 13/42, P < 0.0001). Overall, CCDA is a superior medium compared with CampyBAP for isolating Campylobacter species in our study population. PMID- 10459487 TI - A colorimetric microtiter plate PCR system detects respiratory syncytial virus in nasal aspirates and discriminates subtypes A and B. AB - We developed a colorimetric microtiter plate (MTP) PCR system for specific detection of the respiratory syncytial virus (RSV) nucleocapsid gene and differentiation of viral subtypes A and B. Of 47 pediatric nasal aspirate specimens, the sensitivity and specificity were 94.4% (17 of 18) and 100% (15 of 15), respectively, when compared with RSV cell culture isolation in HEp-2 cells. An additional 14 specimens positive for adenoviruses, rhinoviruses, influenza, or parainfluenza viruses did not give positive reactions. PCR testing detected a mean of 0.15 (0.01 to 7.00) plaque-forming units of RSV virions. Inhibition of PCR amplification was detected in 33.3% (6/18) of undiluted specimens and could be avoided by a dilution (1:10) of extracted RNA without decreasing test sensitivity. RSV subtype, as determined by allele-specific probes, was identical to that determined by an immunofluorescence assay. These results indicate that the MTP PCR system provides a sensitive and specific test for clinical laboratory diagnosis and simultaneous subgroup classification of RSV infection. PMID- 10459488 TI - Interfacial adsorption of polymers and surfactants: implications for the properties of disperse systems of pharmaceutical interest. AB - This review considers basic aspects of the interfacial adsorption of polymers and surfactants, with particular reference to the relevance of these processes for the formulation of pharmaceutical disperse systems. First, we discuss different approaches to the interpretation of adsorption isotherms, paying particular attention to systems containing more than one adsorbate. Second, we consider the implications of adsorption for the properties of suspensions, emulsions, and colloidal systems, particularly as regards the use of polymers and surfactants for stabilizing disperse systems, for controlling flocculation, and for modifying the biopharmaceutical behavior of colloidal drug carriers. Finally, we present a number of representative examples of the importance of adsorption of macromolecules in pharmaceutical systems. PMID- 10459489 TI - Regulatory aspects of stability testing in Europe. AB - The stability data requirements for human pharmaceuticals in the European Community (EC) are based on a series of Directive and Regulation requirements and on a series of advisory guidelines that have been developed and adopted through the International Conference on Harmonization (ICH) procedures or, where there is no relevant ICH project, through the Committee for Proprietary Medicinal Products (CPMP). There are relevant requirements that cover new drugs and finished products containing them and also for existing active ingredients and products containing them. The sources of the relevant information and the data requirements are discussed and summarized. PMID- 10459490 TI - Trends in stability testing, with emphasis on stability during distribution and storage. AB - This paper reviews contemporary trends in the stability testing of pharmaceutical products. In particular, it considers the progress toward globalization and harmonization and indicates stability problems, which probably will be the focus of attention for pharmaceutical scientists and regulators in the near future. Attention is specifically directed to monitoring stability in the channels of distribution. PMID- 10459491 TI - Endogenous and adoptively transferred A-NK and T-LAK cells continuously accumulate within murine metastases up to 48 h after inoculation. AB - In murine models, therapeutic efficacy of adoptive immunotherapy (AIT) of cancer with lymphokine activated killer (LAK) cells is seen only when applied together with substantial doses of interleukin-2 (IL-2), probably because this cytokine is imperative for both motility and viability of the LAK cells. We wanted to investigate whether IL-2 in addition mediates an immunostimulatory activation and expansion of endogenous effector cells contributing to tumor regression. Using an immunoperoxidase technique, we have been able to longitudinally analyze the accumulation of tumor infiltrating lymphocytes expressing the pan-T cell/activated lymphocyte phenotype (Thy1.2), the natural killer (NK) cell phenotype (AsGM,) as well as the cytotoxic T (CD8) cell phenotype within experimental established B16 pulmonary melanoma metastases in C57BL/6 mice during the first 48 h after high dose IL-2 monotherapy. Whereas a substantial and selective infiltration of AsGM1+ lymphocytes in tumor tissue was seen (262 and 937 cells per sq.mm malignant tissue at 0 and 48 h, respectively), only a minor increase in accumulation of CD8+ cells was seen (106 and 171 cells per sq.mm tumor tissue at 0 and 48 h, respectively). The addition of adoptive transfer with lymphokine-activated adherent NK (A-NK) cells to the high-dose IL-2 treatment resulted in more than a 1.5 fold increase in infiltrating AsGM1+ cells compared to IL-2 therapy alone (1520 compared to 937 AsGM1+ cells per sq.mm malignant tissue). No substantial accumulation of CD8+ cells was observed in this setting either. In contrast, the treatment with high dose IL-2 together with adoptive transfer of mitogen-stimulated, lymphokine-activated T killer (T-LAK) cells increased the infiltration of CD8+ cells 10-fold compared to IL-2 monotherapy (2078 compared to 171 CD8+ cells per sq.mm malignant tissue, respectively). Interestingly, infiltration of both endogenous and exogenous cells continued over time, since the effector-to-tumor cell ratio in metastatic tissue dramatically increased from 1:8 and 1:6 at 16 h to 1:3 and 1:2 at 48 h after adoptive transfer of A-NK and T-LAK cells, respectively. These data underline the longevity of LAK cells in vivo and highlight the importance of IL-2 treatment in recruiting endogenous immune cells to tumor areas. PMID- 10459492 TI - Lymphocyte subpopulations anomalies in lung cancer patients and relationship to the stage of disease. AB - BACKGROUND: The host immune response is important in the natural history of neoplastic disease. In order to evaluate alterations of immune function associated with cancer we analyzed the nyctohemeral profile of lymphocyte subpopulations in peripheral blood of cancer patients. The study was carried out on seven healthy volunteers (mean age +/- s.e. 68.8 +/- 1.92), seven patients with I and II stage lung cancer (mean age +/- s.e. 67.2 +/- 0.80), seven patients with III and IV stage lung cancer (mean age +/- s.e. 69.5 +/- 2.26). The area under the curve (AUC) and the presence of circadian rhythmicity were evaluated. RESULTS: The most striking differences were a statistically significant decrease of CD8 (T suppressor/cytotoxic subset) and CD8bright (T suppressor subset) in cancer patients, with a loss of normal circadian rhythmicity, and a statistically significant increase of CD16 (natural killer cells) in cancer patients, especially with I and II stage disease, with a clear circadian rhythm present in all the groups. A statistically significant decrease of delta TcS1 (epitope of the variable domain of delta chain of T-cell receptor 1) was observed in the subjects with I and II stage lung cancer, with a loss of circadian rhythmicity in the two groups of cancer patients. TcR delta 1 (epitope of the constant domain of delta chain of T-cell receptor 1) was significantly decreased in cancer patients, but a clear circadian rhythm was present in these subjects. No significant differences among the groups were found in the values of CD2 (total T cells), CD4 (T helper/inducer subset), CD8dim (T cytotoxic subset), CD4/CD8 ratio, HLA-DR (B cells and activated T cells), CD20 (total B cells) and CD25 (T activated lymphocytes with expression of the alpha chain of interleukin-2 receptor). Nyctohemeral variations of CD2 in control subjects and in I-II stage cancer patients and of CD4 and CD20 in III-IV stage cancer patients presented circadian rhythmicity. CONCLUSIONS: The results suggest that lung cancer is associated with alterations in the proportions and nyctohemeral profiles of various lymphocyte subsets, related to the stage of disease and probably the expression of an altered immune function. PMID- 10459493 TI - Trichloroethylene. I. Carcinogenicity of trichloroethylene. AB - Trichloroethylene (TCE) as an industrial pollutant may damage human health and can be considered as carcinogen. TCE has been detected in the environment and in various human organs, e.g., liver, kidney and brain etc. There are histological alterations such as depletion of glycogen and hydropic degeneration in the liver, however, other signs of TCE effects can be found in various organs as well. TCE and its metabolites, e.g., trichlorethanol, trichloro-acetic acid and epoxides were recently identified as strong mutagens in Ames mutagenicity test inducing frameshift and base-substitution mutations. TCE induced predominantly hepatocellular carcinoma after long term administration in mice. In these animals, kidneys and liver were supposed to be primary target organs with low epoxy-hydrolase activity. A high level of mitotic gene conversion (or gene rearrangement) was indicated by the metabolism of TCE after repeated administration. Purified TCE by was a weak mutagen in the presence of S9 microsomal fraction of rats and as a consequence, the carcinogenic activity was low in the kidney of rats. However, a dose related increase of Leydig cell tumors was found in male rats. PMID- 10459494 TI - Trichloroethylene. II. Mechanism of carcinogenicity of trichloroethylene. AB - The cancer inducing effect of trichloroethylene (TCE) was studied by various methods. DNA complexing activity and apoptosis inhibition were found to be the key elements of the carcinogenicity of TCE and its metabolites. The ability of TCE to interact with DNA was low, but its incorporation into the RNA and DNA of the brain, testis, pancreas, kidney, liver, lung and spleen, cannot be excluded. Exposure to TCE and its metabolites provides a selective growth advantage to spontaneously occurring mutations in some K- and H-ras oncogenes (as non specific results of secondary DNA or RNA damage). The amount of DNA-TCE adducts was higher in mouse hepatocytes than in rat hepatocytes. These differences may explain the species difference in carcinogenicity of TCE, which was dose dependent (due to metabolism) in mice but independent in rats. The blood level kinetics of TCE confirmed the faster metabolic rate in mice, including peroxisome proliferation and induction in hepatocytes. Dichloroacetic- and trichloroacetic acid were found to be hepatic carcinogens in mice, and the specificity depends on peroxisome proliferation induction. Possibly, TCE and related compounds down regulated apoptosis in mouse liver, and the reduced ability to remove initiated cells by apoptosis could be responsible for liver cancer induction by TCE. PMID- 10459495 TI - Trichloroethylene. III. Prediction of carcinogenicity of investigated compounds including trichloroethylene. AB - The mutagenicity and carcinogenic properties of trichloroethylene (TCE) derivatives, and their correlation with its molecular properties were analyzed. The observed cancer incidence was compared to the predicted, calculated incidence. The predictions were based on the rodent bioassay results and were consistent with human data. The electrophilic data of molecules of the Ke system provided evidence for 205 rodent carcinogens, where Ke correlated with energy of the lowest unoccupied molecular orbital. The majority of carcinogenic compounds were found to be electron acceptors with decreased lowest unoccupied molecular orbital (LUMO) energy, indicating the particular DNA-reactivity leading to mutations and abnormal cell division. Based on the mutagenic activity in Ames test, the affinity of target organs for mutagens and non mutagens were compared in 351 rodent carcinogens. Nearly 80% of carcinogens (mutagenic and non mutagenic ones) were positive in the mouse and rat, in at least one of the most frequent target organs, i.e. liver, lung, mammary gland, stomach, kidney, hematopoietic system, urinary bladder and vascular system. Several predictive methods have been developed over the last 5 years based on structure-activity relationship studies known as US National Toxicology Program. One of these programs, called "PROGOL" is widely used for the prediction of carcinogenesis for a wide variety of compounds, e.g., nitro aromatics and suramin analogs. This program provides a simple model for predictive carcinogenesis, despite of the fact that the very first steps of carcinogenesis are not fully understood yet. PMID- 10459496 TI - Influence of theophylline on urinary excretion of total, free and acyl carnitine in rats. AB - BACKGROUND: Carnitine plays a critical role in lipid metabolism. Carnitine deficiency may adversely affect the oxidation of fatty acids and further aggravate abnormal lipid metabolism. OBJECTIVE: To investigate the effect of oral theophylline administration on urinary excretion of total (TC), free (FC), acyl (AC) carnitine as well as the ratio of AC to FC in rats. METHOD: The study was a randomized, controlled animal study. Theophylline was given at 100 mg/kg b.w./day and effects were monitored after a treatment period that lasted between one week and five weeks. RESULTS: Theophylline treatment caused significantly increased food intake and urinary excretion as compared to either control or placebo, P < 0.01. The results indicated that a significant increase in urinary TC, FC and AC excretion as compared to those of control and placebo groups (P < 0.01). Furthermore, the ratio of AC to FC was significantly increased (P < 0.01) as compared to either control or placebo group. CONCLUSION: Theophylline administration to rats leads to significant changes in the urinary excretion of carnitine. These changes may result from theophylline-affected alteration renal tubular re-absorption of carnitine. PMID- 10459497 TI - Lack of the inhibitory effect of intragastrically administered capsaicin on NNK induced lung tumor formation in the A.J mouse. AB - Capsaicin is the principal component in Capsicum fruits consumed by humans worldwide as a food additive. The tobacco-specific nitrosamine 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is thought to be an important causative factor in human lung cancer. Dietary factors are reported to modify lung tumor formation in laboratory animals and in humans. In this study, NNK induced lung tumor formation in female A/J mice given intragastric doses of capsaicin (5 mg/kg body wt) was compared to mice not receiving capsaicin. At the end of the 21 week study, mice treated with capsaicin had an average of 17.1 +/- 1.8 lung tumors/mouse while untreated mice had 19.6 +/- 2. There were 100% lung tumor bearers in each group. Capsaicin alone did not affect spontaneous formation of lung tumors. Our results do not support a possible chemoprotective effect of dietary capsaicin toward NNK-induced lung tumors in human smokers. PMID- 10459498 TI - Evaluation of serological tests for screening of chlamydial eye diseases. AB - Trachoma is recognized as one of most important origins of blindness in developing countries and inclusion conjunctivitis is associated with STD in developed countries. We evaluated the diagnostic value of serological tests for the screening of eye diseases associated with Chlamydia trachomatis infection. We determined serum IgG, IgA and IgM antibodies to C. trachomatis from 53 Japanese patients with active inflammatory trachoma (aged more than 60 years) and from 107 adult patients (aged 20 to 50 years) with acute inclusion conjunctivitis by ELISA test kit. We detected serum IgG antibodies from 22 out of 53 (42.5%) patients with trachoma and from 40 out of 107 (37.4%) patients with acute inclusion conjunctivitis. We also detected serum IgM antibodies from 7 out of 53 (13.2%) patients with trachoma and from 35 out of 107 (32.7%) patients with acute inclusion conjunctivitis. The prevalence of serum IgM antibodies to C. trachomatis in patients with acute inclusion conjunctivitis was significantly higher than that in patients with active trachoma (p < 0.05). Serological tests are also thought to be useful for screening of chlamydial eye diseases. PMID- 10459499 TI - Inhibitory effect of povidone-iodine for the antigen expression of human cytomegalovirus. AB - Anti-human cytomegalovirus (HCMV) properties of povidone-iodine (PVP-I) were evaluated in vitro. The effect of PVP-I was evaluated by indirect immunofluorescence assay on HCMV-infected MRC-5 cells. Percentages of fluorescent cells positive for HCMV immediate early and early antigens in cultures inoculated with AD 169 treated with PVP-I at various concentrations and reaction times were compared with the number of fluorescent cells in the controls inoculated with the virus alone. PVP-I was found to exert some inhibitory effect at a concentration of 0.5% and complete inhibition at a concentration of 7.5% on the infection of MRC-5 cells by HCMV AD 169 strain. Entirely new approaches to the prevention of HCMV infections in infants are necessary. The adequate use of PVP-I as a disinfectant may reduce the transmission of HCMV. PMID- 10459500 TI - Relation between two independent DNA-repair pathways in different groups of naevi. AB - The relation between 2 DNA-repair systems was investigated in 3 groups of naevi (naevus cell naevi, dysplastic naevi, fibromatous naevi) using the correlation coefficient according to Spearman. In the group of naevus cell naevi, only 1 significant correlation between MSH2 and GADD34 expression was found. In the group of dysplastic naevi, 9 significant and highly significant correlations between GADD genes and mismatch repair genes were found. In the group of fibromatous naevi, MLH1 correlated significantly with GADD45 and highly significant with GADD34 expression. Different correlations in naevi groups investigated show the different functional connections between the genes of DNA repair. PMID- 10459501 TI - Growth arrest DNA damage gene expression in naevi. AB - Thirty-one naevus cell naevi, 30 dysplastic naevi and 12 fibromatous naevi were stained for the presence of p53 and Growth Arrest DNA Damage genes. All naevus cell naevi and fibromatous naevi were highly positive for GADD genes and negative for p53. Dysplastic naevi had significantly lower GADD34 and GADD153 expression as well as higher p53 expression in relation to the other naevi groups. The absence or decrease of GADD genes expression in naevus may indicate a potential malignant transformation. PMID- 10459502 TI - Expression of DNA mismatch repair genes in naevi. AB - The significant difference of DNA mismatch repair genes expression between naevi and melanomas was demonstrated by our research group in the previous study. The main aim of this study was to compare the expression of MLH1, MSH2, PMS1 and PMS2 in 31 naevus cell naevi, 12 fibromatous naevi and 30 dysplastic naevi. The expression of DNA mismatch repair proteins was found in all naevi investigated. However, the expression (percentage of positively stained cells) was significantly higher in naevus cell naevi than in dysplastic and fibromatous ones. PMID- 10459503 TI - Indomethacin reduces the skin thermal damage in hyperthermic treatment of experimental malignant tumors. AB - Indomethacin (Ind), an inhibitor of prostaglandin synthesis, was previously shown to increase the thermosensitivity of murine tumors. The potency of drug to modify the thermal response of murine skin has been evaluated in mice heated in water bath at 44 degrees C for 30, 60 and 90 min. Ind was administered subcutaneously (s.c.) at dose of 5 mg/kg body weight (b.w.) 1.5 h before heating. The mouse foot skin reactions (FSR) were assessed using the scoring system of Urano et al (1979). The severity of skin thermal damage was decreased markedly by Ind. At the time when heating of the control group at 44 degrees C for 60 min resulted in the irreversible FSR in some mice, pretreatment with Ind before heating lead to the complete recovery from the heat damage in all mice. Similarly, after heating at 44 degrees C for 90 min, the degree of FSR was diminished by Ind from score 4.5 to score 2. It is concluded that Ind selectively protected normal skin during the hyperthermic treatment. Further clinical study is warranted. PMID- 10459504 TI - Diazepam treatment in rats induces changes in the concentrations of different phospholipid classes in liver and liver mitochondria. AB - Liver phospholipid concentrations were determined in rats after the administration of diazepam (5 mg/Kg/day), for a period of two months. Increased concentrations of total phospholipids (P < 0.05), phosphatidylcholine (P < 0.05) and phosphatidylinositol (P < 0.05) were found in the rats taking diazepam. In contrast, a decreased concentration of phosphatidylserine (P < 0.01) was observed in the same group of animals. In addition, changes in the concentration of rat liver mitochondrial phospholipids after the administration of diazepam during the same period of time were determined. Increased concentrations of total phospholipids (P < 0.01), phosphatidylcholine (P < 0.001) and diphosphatidylglycerol (P < 0.001) were found in the rats treated with diazepam. In contrast, decreased phosphatidylserine (P < 0.001) and phosphatidylinositol (P < 0.01) concentrations were observed in the same group of animals. The considerable changes observed in liver phospholipids and individual classes of liver mitochondrial phospholipids induced by long-term administration of diazepam, possibly suggest a stimulation of liver phospholipid biosynthesis. This effect may be related to enzymatic systems which are involved in phospholipid pathways, and are linked to benzodiazepinergic binding sites. PMID- 10459505 TI - Decreased retinoblastoma protein expression in gamma-irradiated mouse ovarian follicles. AB - We immunohistochemically examined the effect of gamma-radiation on immature mouse ovarian follicles. Mice were Jirradiated with a dose of LD80 (8.3 Gy) in KAERI. At 0 h, 6 h, 12 h, 1 d, 2 d, 4 d and 8 d postirradiation, the ovaries were excised and fixed in neutral buffered formalin. We performed immunohistochemistry for protein retinoblastoma (pRb), terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling (TUNEL) and routine hematoxylin-eosin staining in the largest cross sections. Radiation-induced follicular degeneration increased before 6 h, and most irradiated ovarian follicles became acutely atretic. The immunohistochemical staining for pRb was strong in the nuclei of granulosa cells of normal follicles and weak in atretic ones which were, conversely, strong for TUNEL staining. It was shown that pRb expression became lower with the degeneration of the ovarian follicles, which was inhibited by gamma-radiation. In the present study, pRb immunohistochemistry was proven to be a useful tool for the identification of follicular status. PMID- 10459506 TI - Molecular biological ontogenesis of the thymic reticulo-epithelial cell network during the organization of the cellular microenvironment. AB - The thymus provides an optimal humoral microenvironment for the development of immunocompetent T cells. Although yolk sac derived pre-T, committed hematopoietic stem cells enter the thymus using a homing receptor, the immigration process also requires secretion of a peptide called thymotaxin by the cells of the reticulo epithelial (RE) network of the thymic cellular microenvironment. The majority of RE cells have a round or irregular pale nucleus, which contains few, scattered, chromatin granules with a defined, spherical nucleolus, rich in basic histones. Their cytoplasm occasionally displays RNP granules, and is rich in non-histone proteins, fine phospholipid, lipid or cholesterin granules, and vacuoles filled with secreted substances. The cells of the subcapsular, endocrine RE cell layer (giant or nurse cells), characterized by PAS positive granules, express A2B5/TE4 cell surface antigens and MHC Class I (HLA A, B, C) molecules. In contrast to medullar RE cells, these subcapsular nurse cells also produce thymosins beta 3 beta 4. Thymic nurse cells (TNCs) display a neuroendocrine cell specific immunophenotype (IP): Thy-1+, A2B5+, TT+, TE4+, UJ13/A+, UJ127.11+, UJ167.11+, UJ181.4+, and presence of common leukocyte antigen (CLA+). Medullar RE cells display MHC Class II (HLA-DP, HLA-DQ, HLA-DR) molecule restriction. These cells also contain transforming growth factor-beta (TGF-beta) type II receptors and participate in the positive selection of T cells. Transmission electron microscopic (TEM) observations have defined four functional subtypes of medullar RE cells: undifferentiated, squamous, villous, and cystic. All subtypes are connected by desmosomes. Immunocytochemical observations have shown that the secreted thymic hormones, thymosin alpha 1 and thymopoietin (and its short form, thymopentin or TP5), are produced by the same RE cells. Thymic RE cells also produce numerous cytokines including IL1, IL6, G-CSF, M-CSF, and GM-CSF that likely are important in various stages of thymocyte activation and differentiation. The co-existence of pituitary hormone and neuropeptide secretion, such as growth hormone, prolactin, adrenocorticotropic hormone, thyroid stimulating hormone, triiodothyronine, somatostatin, oxytocin, follicle stimulating hormone, luteinizing hormone, arginine vasopressin, growth hormone releasing hormone, corticotropin releasing hormone, nerve growth factor, vasoactive intestinal peptide, (pro) enkephalin, and beta-endorphin, production of a number of interleukins and growth factors, as well as the expression of receptors for all, by the same RE cell is an unique molecular biological phenomenon. These data illustrate the immensely important and diverse immuno neuroendocrine functions of the thymic RE cellular network. Based on our systematic observations of the thymus in humans and other mammalian species, we suggest that the thymic RE cell network represents an extremely important cellular and humoral microenvironment in homeopathic regulatory mechanisms of the multicellular organism. Intrathymic T lymphocyte selection is a complex, multistep process, influenced by several functionally specialized RE cell subtypes and under constant immuno-neuroendocrine regulation, reflecting the dynamic changes of the organism. PMID- 10459507 TI - Lipid peroxidation and tissue damage. AB - In recent years it has become apparent that the oxidation of lipids, or lipid peroxidation, is a crucial step in the pathogenesis of several disease states in adult and infant patients. Lipid peroxidation is a process generated naturally in small amounts in the body, mainly by the effect of several reactive oxygen species (hydroxyl radical, hydrogen peroxide etc.). It can also be generated by the action of several phagocytes. These reactive oxygen species readily attack the polyunsaturated fatty acids of the fatty acid membrane, initiating a self propagating chain reaction. The destruction of membrane lipids and the end products of such lipid peroxidation reactions are especially dangerous for the viability of cells, even tissues. Enzymatic (catalase, superoxide dismutasse) and nonenzymatic (vitamins A and E) natural antioxidant defence mechanisms exist; however, these mechanisms may be overcome, causing lipid peroxidation to take place. Since lipid peroxidation is a self-propagating chain-reaction, the initial oxidation of only a few lipid molecules can result in significant tissue damage. Despite extensive research in the field of lipid peroxidation it has not yet been precisely determined if it is the cause or an effect of several pathological conditions. Lipid peroxidation has been implicated in disease states such as atherosclerosis, IBD, ROP, BPD, asthma, Parkinson's disease, kidney damage, preeclampsia and others. PMID- 10459508 TI - Psychiatric labelling, sex role stereotypes and beliefs about the mentally ill. AB - This study aimed to investigate the effect of two main variables on specific attitudes and beliefs about mental illness. The first variable tested the labelling theory proposal that the label of mental illness per se is stigmatizing for those so labelled. The second tested the proposal of Rosenfield (1982) that males and females receive a more severe societal reaction for deviance when the deviant behaviour is inconsistent with traditional sex role norms. Questionnaires with vignettes describing four behaviour types were given to young adult respondents. The expected effect of a psychiatric diagnosis and of deviance from sex role stereotypes were not confirmed. It was concluded that while several variables combine to influence specific attitudes and beliefs about the mentally ill, the type of behaviour displayed is the crucial factor. PMID- 10459509 TI - Social networks and mental health service utilisation--a literature review. AB - Social networks have been shown to be smaller in individuals with severe mental illness than in the general population. Patients' social networks and social support may impact on their utilisation of psychiatric services. This literature review focuses on social networks, social support and mental health service use in patients with mental illness. Most studies suggest that smaller social networks or less social support are associated with more frequent hospitalisation. Qualitative aspects of the social network are also related to the risk of hospitalisation. The relationship between social networks and other types of service use is not established. PMID- 10459510 TI - Suicides in Hong-Kong and Singapore: a tale of two cities. AB - Suicide statistics from Hong Kong and Singapore over the period 1984-1994 were studied. The suicide behaviours in Hong Kong and Singapore are remarkably similar. Suicide rates increased with age, with a sharp increase among the elderly which was about four to five times the average. A relatively low male:female ratio and low teenage suicide were also found. Jumping from a height was the commonest method of suicide. Nevertheless, a different time trend of the suicide rates was observed, with an upward and downward trend for Hong Kong and Singapore respectively. The cultural, social and economic aspects in understanding suicidal behaviour in Hong Kong and Singapore are also discussed. PMID- 10459511 TI - Suicidality and personality in American and Kuwaiti students. AB - In samples of American and Kuwaiti college students, current and prior suicidal preoccupation were associated with measures of psychological disturbance, with depression being the strongest predictor of suicidality in both samples. Although the Kuwaiti students were more depressed than the American students, they reported less prior suicidal preoccupation. PMID- 10459512 TI - Predicting psychological distress among former Soviet immigrants. AB - This study investigated whether age, gender, marital status, education, employment, length of time in the U.S. and immigration demands (novelty, occupation, language, discrimination, loss, and not feeling at home) were predictors of psychological distress in a sample of 1,647 former Soviet immigrants. Multiple regression analysis indicated that the combined model of demographic and demand of immigration variables was significant. Results indicated that women, older immigrants, those with less than a college education, and those with greater immigration demands related to novelty, language, discrimination, loss, and not feeling at home were most distressed. PMID- 10459513 TI - Measuring consumer participation in mental health services: are attitudes related to professional orientation? AB - This study describes the development of the Consumer Participation Questionnaire to measure consumer involvement in the planning, management and evaluation of mental health services, and the attitudes of mental health workers towards consumer participation. Results indicate that while most professionals view the concept positively, progress may be occurring faster at the level of individual treatment than at the organisational level. Professionals with a more biological (as opposed to psychosocial) orientation were less likely to predict that services would improve if consumers were involved in the planning of services or were employed therein. Recommendations to facilitate more effective consumer participation are made, including the need for the value of a collaborative approach to be emphasised in professional training programmes. PMID- 10459514 TI - A law affecting medically assisted procreation is on the way in Switzerland. PMID- 10459515 TI - A simplified coculture system using homologous, attached cumulus tissue results in improved human embryo morphology and pregnancy rates during in vitro fertilization. AB - PURPOSE: This study was undertaken to evaluate simplified methods of human embryo coculture using either attached or nonattached autologous cumulus tissue. METHODS: Eight hundred one zygotes were cultured for 48 hr in a prospective, randomized trial comparing culture of embryos either with intact cumulus tissue, with cumulus tissue added to the droplet of culture medium, or without any cumulus tissue. In a follow-up study, embryo quality, pregnancy rates, and implantation rates were compared in 120 consecutive patients undergoing in vitro fertilization with a coculture system using cumulus tissue compared to a cohort of 127 patients undergoing IVF immediately preceding the institution of the coculture protocol. RESULTS: Embryo morphology was significantly improved (P < 0.05) following culture with attached cumulus tissue (5.61 +/- 0.29) and culture with added cumulus tissue (4.72 +/- 0.31) compared to that of embryos grown in culture medium without cumulus tissue (3.95 +/- 0.26). The clinical pregnancy rate improved from 39.4% (50/127) to 49.2% (59/120) following institution of a system of coculture with attached cumulus tissue. CONCLUSIONS: These data indicate that a simple coculture system using autologous cumulus tissue can result in improved embryo morphology, implantation rates, and clinical pregnancy rates during in vitro fertilization. This coculture system is simple, is non labor intensive, and eliminates many of the risks which may be present in other embryo coculture systems. PMID- 10459516 TI - The effect of hydrosalpinges on IVF-ET outcome. AB - PURPOSE: Our purpose was to determine if the presence of a hydrosalpinx effects the outcome of in vitro fertilization (IVF)-embryo transfer. METHODS: We performed a retrospective analysis of IVF cycle stimulation sheets. RESULTS: A total of 1000 patients with tubal factor infertility was analyzed. There were 60 hydrosalpinx patients who underwent 116 initiated cycles with 106 embryo transfers, compared to 940 control patients undergoing 1428 initiated cycles with 1150 embryo transfers. Both groups had a similar response to ovarian stimulation, number of oocytes retrieved, and number of embryos transferred. The hydrosalpinx group had a significantly higher preclinical loss rate (22/59 = 37% vs 80/566 = 14%; P = 0.001), a significantly lower implantation rate (55/352 = 16% vs 795/3795 = 21%; P = 0.013), a trend toward a reduced delivery rate per transfer (28/106 = 26% vs 387/1150 = 34%; P = 0.066), a significantly higher ectopic pregnancy rate (5/59 = 8% vs 16/566 = 3%; P = 0.04), and a similar spontaneous abortion rate (9/37 = 24% vs 99/486 = 20%; P = 0.28) compared to the control tubal factor group. CONCLUSIONS: This study demonstrates a decrease in implantation rates and an increase in preclinical miscarriages and ectopic pregnancies in patients with hydrosalpinges compared to tubal-factor patients without sonographic evidence of dilated fallopian tubes. PMID- 10459517 TI - Fertilization and development of a blastocyst-stage embryo after selective intracytoplasmic sperm injection of a mature oocyte from a binovular zona pellucida: a case report. AB - PURPOSE: Our purpose is to describe the development of a blastocyst-stage embryo after the selective fertilization of a mature oocyte from a binovular zona pellucida by intracytoplasmic sperm injection (ICSI). METHOD: A 34-year-old woman underwent intracytoplasmic sperm injection due to severe male-factor infertility. After oocyte retrieval, a binovular zona pellucida including one mature metaphase II oocyte and one immature oocyte at the germinal vesicle stage as well as nine metaphase II oocytes was injected with spermatozoa using a one-to-one approach. RESULTS: The injected mature oocyte of the binovular zona pellucida showed fertilization as evidence by the presence of two pronuclei and cleaved to a four cell embryo on the second day, while the uninjected oocyte showed signs of degeneration. On the third day, this embryo further cleaved to six blastomeres with slight fragmentation and it reached the blastocyst stage on the sixth day. CONCLUSIONS: Selective fertilization of one oocyte from a binovular zona pellucida by ICSI may lead to the development of a morphologically normal blastocyst-stage embryo which can be used for embryo transfer in the presence of a limited number of embryos. PMID- 10459518 TI - Comparison of pregnancy outcome of pronuclear- and multicellular-stage frozen thawed embryo transfers. AB - PURPOSE: Our purpose was to determine if supernumerary embryos generated by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) should be frozen (using 1,2-propanediol) at the pronuclear or multicellular stage. METHODS: The study was a retrospective analysis conducted at the Dubai Gynaecology & Fertility Centre of the Department of Health & Medical Services, Dubai, U.A.E. One hundred forty-one women undergoing frozen-thawed embryo replacement cycles with IVF generated embryos and 84 women undergoing the same with ICSI generated embryos. RESULTS: Supernumerary, IVF-generated embryos frozen at the multicellular stage had a significantly higher rate of survival on thawing (73.9%) than embryos frozen at the pronuclear stage (64.4%). The morphological grades of the embryos in the two groups were similar, but a significantly higher pregnancy rate was obtained with embryos frozen at the multicellular stage (22.8%) than with pronuclear-stage embryos (14.8%). Similarly, with ICSI generated embryos, significantly higher survival was seen with multicellular stage frozen embryos (74.8%) than pronuclear-stage embryos (64.4%). The morphological grades of the embryos and pregnancy outcomes of the two groups were similar. CONCLUSIONS: Supernumerary embryos generated by IVF and ICSI should be frozen at the multicellular stage so as to allow selection of the best embryos for transfer and embryo freezing of only robust embryos. PMID- 10459519 TI - Exogenous luteinizing hormone (LH) increases estradiol response patterns in poor responders with low serum LH concentrations. AB - PURPOSE: Our purpose was to investigate whether the addition of exogenous leuteinizing hormone (LH) increases estradiol secretion in LH-depleted women undergoing controlled ovarian hyperstimulation (COH) with purified follicle stimulating hormone (FSH). METHODS: We carried out case series and retrospective analysis of midfollicular serum LH concentrations and estradiol response patterns in COH cycles. All patients initially received gonadotropins containing purified FSH. Human menopausal gonadotropin containing LH was added to poor responders with low midfollicular LH concentrations. RESULTS: The addition of exogenous LH to the COH regimen significantly increased estradiol secretion in poor responders with low midfollicular endogenous LH concentrations. This was confirmed statistically by an average change in the slope of the estradiol patterns from 27.54 to 85.49 after the addition of exogenous LH. Furthermore, patients with midfollicular serum LH concentrations < 3.0 mIU/ml had significantly lower midfollicular and peak estradiol (E2) concentrations compared to patients with LH concentrations > or = 3.0 mIU/ml (352.3 and 2094.3 vs 855.6 and 3757.1 pg/ml, respectively). CONCLUSIONS: The addition of exogenous LH increases E2 response patterns in poor responders with low midfollicular serum LH concentrations. Low midfollicular serum LH concentrations are associated with significantly lower midfollicular and peak E2 concentrations. PMID- 10459520 TI - Effect of a sonographically diffusely enlarged uterus without distinct uterine masses on the outcome of in vitro fertilization-embryo transfer. AB - PURPOSE: Our objective was to study the effect of a sonographically diffusely enlarged uterus without distinct uterine masses on the outcome of in vitro fertilization-embryo transfer (IVF-ET). METHODS: Nineteen primary infertility patients undergoing IVF-ET who had a sonographically diffusely enlarged uterus without distinct uterine masses were enrolled. An age-controlled group of 144 primary infertility patients undergoing IVF-ET with a normal uterus and no history of uterine surgery was included. RESULTS: The age, day 3 follicle stimulating hormone, antral follicle count, ovarian response, endometrial thickness, number of retrieved and fertilized oocytes, number of transferred embryos, clinical pregnancy rate, and total delivery rate were not statistically different between the two groups (P < 0.05). Patients with a sonographically diffused enlarged uterus without distinct uterine masses had a higher spontaneous abortion rate (66.7%) than controls (P < 0.04; odds ratio = 7.5; 95% confidence interval, 1.16-48.56). CONCLUSIONS: A high spontaneous abortion rate was found in patients with a sonographically diffusely enlarged uterus without distinct uterine masses undergoing IVF-ET. Enhanced luteal support was required. PMID- 10459521 TI - Fimbrial capture of the ovum and tubal transport of the ovum in the rabbit, with emphasis on the effects of beta 2-adrenoreceptor stimulant and prostaglandin F2 alpha on the intraluminal pressures of the tubal ampullae. AB - PURPOSE: Our purpose was to elucidate the roles of the ampullar and isthmic portions of the oviduct and the effects of drugs on oviductal contractility. METHODS: Prostaglandin F2 alpha (PGF2 alpha; Ono Pharmaceuticals, Osaka) and oxytocin (Atonin-O; Teikoku Hormone Manufacturing Co. Ltd., Tokyo) were used to stimulate oviductal contractility, and ritodrine hydrochloride (Utemerin; Solvay Duphar Corp., Denmark) to inhibit the contractility. RESULTS: Both PGF2 alpha and Atonin-O were involved in ovum capture by the ampullar oviduct by stimulating contractility, thus altering the intraductal pressures. Utemerin is effective in inhibiting the enhanced contractility induced by PGF2 alpha and Atonin-O. CONCLUSIONS: Variations in pressure of the ampullar portion of the oviduct seem necessary for the capture of ova expelled from the ovary. Once in the isthmic portion of the oviduct, transport appears to be under the influence of ciliary activity rather than variations in contractility. PMID- 10459522 TI - Effect of Vero cell coculture on the development of frozen-thawed two-cell mouse embryos. AB - PURPOSE: Our purpose was to evaluate the beneficial effects of long-term coculture of Vero cells on the development of frozen-thawed two-cell mouse embryos. METHODS: Two-cell mouse embryos were frozen slowly with 1,2-propandiol and sucrose as cryoprotectants and thawed rapidly, followed by stepwise dilution. Vero cells were cultured in drops of RPMI 1640 to establish monolayers. Frozen thawed embryos were cultured alone (control) or cocultured with Vero cells. The rate of development in both groups was compared. RESULTS: After 4 days of culture, significantly more embryos in coculture were developed to expanded blastocysts (61 vs 37% for controls; P < or = 0.0001). In addition, on the fifth day of cultivation, more embryos in coculture showed the potential of hatching from the zona pellucida (26 vs 7% in controls; P < or = 0.0001). The rate of degeneration in coculture was also much lower than in controls (6 and 15%, respectively). CONCLUSIONS: Coculture of cryopreserved preimplantation-stage embryos with Vero cells seems to be a useful tool to eliminate the postthaw deleterious effect of freezing and also to obtain better-quality embryos appropriate for transfer. PMID- 10459523 TI - Unequal pronuclear size--a powerful predictor of embryonic chromosome anomalies. AB - PURPOSE: Our purpose was to evaluate whether pronuclei of unequal size, observed in 13.7% of zygotes evaluated after in vitro fertilization (IVF), are predictive of chromosome anomalies in the developing embryo. METHODS: Five ploidy of 38 embryos grown from zygotes with unequal-sized pronuclei was assessed by fluorescent in situ hybridization (FISH). Twenty-six embryos developed after intracytoplasmic injection of sperm (ICSI) and 12 embryos were derived from conventional IVF. RESULTS: Chromosome anomalies were documented in the ICSI and IVF groups in 88.5 and 50% of cases, respectively. CONCLUSIONS: We suggest that FISH should be employed to examine the ploidy of zygotes with unequal pronuclei, prior to embryo transfer. PMID- 10459524 TI - Intraoperative applications of saline infusion ultrasonography. PMID- 10459525 TI - Interleukin-1 beta production by human preimplantation embryos. PMID- 10459526 TI - Intratester and intertester reliability of goniometric measurement of passive lateral shoulder rotation. AB - Measurement of lateral rotation range of motion (ROM) is frequently performed during shoulder evaluation. The purpose of this study was to determine the intratester and intertester reliability of goniometric measurement of passive lateral rotation ROM of the shoulder. Two experienced PTs performed the testing in a randomized block design. They were blinded to all clinical information and to their goniometric readings. Passive lateral rotation ROM of the shoulder was assessed in 34 patients with a variety of shoulder pathologies. Patients were placed in the supine position with the arm abducted approximately 20 degrees to 30 degrees. A standard goniometer, placed along the joint axis by the therapist, was red by an independent assistant. Intraclass correlation coefficients (ICCs) and their associated 95% confidence intervals were calculated. Intratherapist ICCs (0.88 and 0.93) and intertherapist ICCs (0.85 and 0.80) were high. These findings suggest that reliable measures of passive lateral rotation ROM of the shoulder can be obtained from patients with shoulder pathology using standard goniometry and by placing the patient in a supine position. PMID- 10459527 TI - Maximum grip strength in normal subjects from 20 to 64 years of age. AB - The purposes of this study were to develop normative maximum grip strength (GRIP) data for men and women aged 20 to 64 years, separated into nine five-year age groups, and to develop prediction equations for GRIP using gender, age, height, weight, and hand dominance. A total of 1,182 volunteers (553 men and 629 women) participated in the study. Maximum hand grips were obtained using the Jamar dynamometer with standardized positioning and instructions. The hand to be tested first was chosen randomly. Each hand was then tested alternately. Three trials were performed on each hand. The highest GRIP for each hand was used for analysis. Two-way analyses of variance showed significant differences between the right and left hands and across the age groups for both genders. Follow-up analyses showed that significant decreases occurred between the age groups of 50 54 and 55-59 years in men and between the age groups of 50-54 and 60-64 years in women. The data also indicated that right and left GRIPs were highly correlated with each other (r = 0.93). Gender, height, and weight moderately correlated with both GRIPS (r = 0.52-0.73. Age correlated weakly with both GRIPs (r = -0.17). If either the right or the left GRIP was known, the other GRIP could be predicted easily from the known GRIP, with 87% of variance accounted for. Without the knowledge of the other GRIP, either GRIP could be predicted through gender, height, weight, age, and hand dominance, with 61% to 62% of the explained variance. The norms and prediction equations of GRIP developed in this study for men and women aged 20 to 64 years will help clinicians with decision making regarding grip strength. PMID- 10459528 TI - Functional limitation immediately after cast immobilization and closed reduction of distal radius fractures: preliminary report. AB - The majority of research on distal radius fractures consists of retrospective, descriptive studies of patients with unstable fractures requiring fixation. The purpose of this investigation was to report on impairments in flexibility, grip strength, and motor control and on the presence of swelling and atrophy immediately after cast immobilization of closed reductions of simple distal radius fractures. Sixteen adult subjects from Kaiser Permanente Medical Center, San Francisco, entered the study, and 13 completed it. At the initial evaluation, upper extremity ranges of motion, grip strength, forearm circumferences, two point discrimination, and motor reaction times were measured on the uninvolved side. The same measurements were taken on the affected side within 48 hours after cast removal. All but one subject worked throughout the casting period. There were significant postcasting impairments in forearm rotation (40% deficit in pronation and supination); wrist flexion, extension, and radial and ulnar deviation (50% reduction in all motions); grip strength (-32 kg, or approximately 24% of the strength of the unaffected side); and forearm circumference (-1.1 cm) and wrist circumference (+1.5 cm). Patients complained of awkwardness of the involved hand. These measured impairments immediately after immobilization of simple radius fractures were greater than the reported impairments in patients after reduction of radius fractures with fixation 6 to 27 months after injury. To prevent long-term disability and recover flexibility, strength, and function, patients with simple distal radius fractures should be referred to a hand, occupational, or physical therapist for evaluation, education, and treatment after immobilization. Longitudinal studies are needed to quantify long-term functional recovery with regard to the type of fracture and the degree of impairment measured immediately after casting. PMID- 10459529 TI - Dynamic versus static grip strength: how grip strength changes when the wrist is moved, and why dynamic grip strength may be a more functional measurement. AB - The synergistic relationship between wrist/forearm range of motion (ROM) and grip strength (GS) is arguably one of the most important aspects of hand function. Clinically, GS is measured with the wrist in a standardized static position, and the results of such tests have been deemed valid and reliable. The question remains, however, whether this static GS (SGS) measurement is an accurate indication of how an individual functionally grips objects--that is, most functional tasks require the fingers to grasp an object forcibly while moving the proximal joints such as the wrist and forearm. Therefore, further analysis of an individual's dynamic GS (DGS) during wrist/forearm movements may improve the clinician's understanding of hand function and provide more pertinent guidelines for assessing functional gripping, e.g., for vocational and avocational tasks and in designing workstations. The purpose of this study is twofold: to describe and assess a DGS testing device that utilizes optically encoded gyroscopes and a strain-gauge dynamometer to simultaneously measure GS and wrist/forearm position over real time; and to assess and compare grip force production differences in SGS and DGS in uninjured wrists, using this novel device. Twenty-nine uninjured wrists of men (n = 15) and women (n = 14)--age range, 21 to 43 years--were tested with the DGS device. Subjects were excluded if they had any previous wrist/forearm fracture, pain, or limitation of motion. The DGS device was designed and fabricated with two optically encoded gyroengines, a vertical gyroscope with two axes for measuring flexion/extension and radial/ulnar deviation, and a directional gyroscope with one axis for measuring supination/pronation, mounted on a strain-gauge dynamometer. The signals from the gyroscopes and dynamometer were processed by means of a data aquisition board and analog-to-digital circuitry and collected on a 486-MHz computer. The methods included repeated testing of each gyroscope axis to known angular measurements, and randomly assigned maximal gripping trials from the 29 subjects. The standard deviation of gyroengines is 1 degree to 2 degrees for each ROM axis. Maximal DGS is significantly less (mean 14%) than SGS, and SGS is 29% less than DGS at the same three-dimensional ROM positions. Gyroengines are feasible three-dimensional tracking devices that can be used to monitor wrist/forearm ROM in conjunction with GS. PMID- 10459530 TI - Management of a metacarpophalangeal joint fracture using a dynamic traction splint and early motion. AB - Dynamic traction and early motion have been used by hand therapists to treat proximal interphalangeal joint fractures with good results. However, metacarpophalangeal joint fractures have been neglected, perhaps because of their relative infrequency. The purpose of this case study is to report the authors' experience using dynamic traction in conjunction with early motion to treat a construction worker who fell from scaffolding and sustained a complex fracture of the left smallfinger proximal phalanx with involvement of the metacarpophalangeal joint. Range of motion of the patient's left small finger at discharge from therapy, approximately 22 weeks postoperatively, was as follows: metacarpophalangeal joint, hyperextension/65 (85 degrees passive flexion); proximal interphalangeal joint, 20/80 (5/90 passively); and distal interphalangeal joint, 0/50 (65 degrees passive flexion). The patient made a successful return to full-time construction work. The results of this case appear to support consideration of the use of dynamic traction and early motion for management of selected metacarpophalangeal joint fractures. PMID- 10459531 TI - Making hinges with thermoplastic tubing. PMID- 10459532 TI - Case reports and case studies: a discussion of theory and methodology in pocket format. AB - Issues of the theory and methodology of case reports and case studies have been presented. Examples of applications of techniques used in the rehabilitation of the upper extremity conclude this report in pocket format on the theory and methodology of case reports and case studies. Case reports and studies enhance our knowledge about practice in ways that are unique to each case, which are not possible when investigations are performed through group comparison. Case inquiries help us share our practice, sharpen our theoretical reasoning, develop our good habits of applying reliable and valid measurements, and above all help us explore and discuss our treatments. PMID- 10459533 TI - Porcine islet isolation: a real advance? PMID- 10459534 TI - N-acetylcysteine in pig liver transplantation from non-heart-beating donors. AB - Lipid peroxidation due to oxygen free radicals (OFR) seems to play a major role in loss of liver graft viability after warm ischemia, preservation, and transplantation. N-acetylcysteine (NAC) is an antioxidant that has a direct effect on OFR, and is also a glutathione precursor, another antioxidant. This study was designed to evaluate the efficacy of NAC in preventing ischemia reperfusion damage of liver grafts harvested from non-heart-beating donors. Liver transplantation was performed on pigs divided into five groups: group 1 (control group; n=5) received livers from heart-beating donors; livers were subjected to 30 min of warm ischemia in groups 2 (n=3, no NAC) and group 3 (n=3; NAC treatment); warm ischemia time lasted 60 min in groups 4 (n=4; no NAC) and 5 (n=5; NAC treatment). Studied parameters included graft survival for more than 3 days, aspartate aminotransferase plasma levels, liver histology, and hepatic total glutathione concentrations. Graft survival was 100% in groups 1, 2, and 3, 0% in group 4, and 20% in group 5. NAC treatment did not influence initial mean aspartate aminotransferase release which was greater in warm ischemic livers than in controls. NAC treatment had no effect on liver hepatic total glutathione after reperfusion of animals receiving warm ischemic grants. Finally, no effect on liver histology was observed with NAC treatment. Our study suggests that in liver transplantation from non-heart-beating donors, NAC has no effect in both graft viability and lipid peroxidation. The role of OFR in primary dysfunction of transplanted warm ischemic livers remains controversial. PMID- 10459535 TI - Induction, maintenance, and reversal of streptozotocin-induced insulin-dependent diabetes mellitus in the juvenile cynomolgus monkey (Macaca fascilularis). AB - BACKGROUND: Insulin-dependent diabetes mellitus (IDDM) is the second most prevalent chronic illness of children. Investigation of the treatment of IDDM is hindered by the lack of a reproducible and easily maintained non-human primate model of this disorder. METHODS: We induced IDDM in 11 juvenile cynomolgus monkeys after a single (150 mg/kg) intravenous injection of streptozotocin (STZ). All diabetic monkeys were treated with insulin twice daily, based on a sliding scale. Subcutaneous vascular access ports were surgically placed in each monkey to facilitate serial blood sampling and drug administration. Allogeneic pancreatic islet cells from unrelated donors were subsequently transplanted into the mesenteric circulation of all STZ-treated monkeys. RESULTS: Mild, transient nausea and vomiting occurred in all animals after STZ injection; however, no additional signs of toxicity occurred. Within 36 hr, all monkeys required twice daily administration of exogenous insulin to maintain a non-ketotic state. Serum C-peptide levels decreased from >1.2 ng/ml before STZ, to between 0.0 and 0.9 ng/ml after STZ, confirming islet cell destruction. Animals were maintained in an insulin-dependent state for up to 147 days without any observable clinical complications. Subcutaneous vascular access port patency was maintained up to 136 days with a single incidence of local infection. Islet cell transplantation resulted in normoglycemia within 24 hr. Serum C-peptide levels increased (range: 2-8 ng/ml) for 6 - 8 days in immune competent animals, and for 39-98 days after transplant in immunosuppressed monkeys. CONCLUSIONS: IDDM can be consistently induced and safely treated in juvenile cynomolgus monkeys. Chronic vascular access can be maintained with minimal supervision and complications. This model is appropriate for studies investigating potential treatments for IDDM including islet cell transplantation. PMID- 10459536 TI - CD34+ selected bone marrow grafts are radioprotective and establish mixed chimerism in dogs given high dose total body irradiation. AB - BACKGROUND: Canine stem cell transplantation models have provided important preclinical information for human clinical studies. The recent cloning of cDNA for canine CD34 and the production of monoclonal antibodies that recognize canine CD34 have been the basis for the development of techniques for the large-scale enrichment of canine hematopoietic progenitor cells. In this study, we evaluated the in vivo functional properties of canine bone marrow CD34+ cells after a myeloablative conditioning regimen. METHODS: After 920 cGy total body irradiation, three dogs received infusion of autologous CD34+ selected cells from the marrow, three dogs CD34+ depleted autologous marrow cells, and two dogs received CD34+ autologous marrow cells that were immunomagnetically selected and then further purified by cell sorting. In addition, four dogs received allogeneic marrow enriched for CD34+ cells from dog leukocyte antigen-identical littermates to investigate long-term repopulating function of CD34+ cells. Chimerism studies were performed using polymerase chain reaction to detect highly polymorphic microsatellite markers. RESULTS: In three recipients of autologous marrow enriched for CD34+ cells to between 29% and 70% (1.6 x 10(6) to 3.4x10(6) CD34+ cells/kg), prompt and full hematopoietic recovery occurred, whereas in three dogs that received marrow depleted of CD34+ cells (1 x 10(7) cells/kg), no hematopoietic recovery was achieved. In two dogs that received highly purified CD34+ cells (purity: 98% and 96%, 0.79x10(6) to 0.547x 10(6) CD34+ cells/kg), delayed but full hematopoietic recovery was seen. Three of four allograft recipients of 1.75x10(6) to 6.8x10(6) CD34+ cells/kg engrafted and showed full hematopoietic recovery, whereas one dog rejected the graft. The three long-term survivors showed stable mixed hematopoietic chimerism with predominantly donor hematopoiesis. CONCLUSION: Transplantation of canine CD34+ cells after lethal total body irradiation provides radioprotection and gives rise to long-term hematopoietic reconstitution. Stable donor/host mixed chimerism was observed in allograft recipients most likely as a result of T-cell depletion of the grafts. Our findings suggest a future role for canine preclinical transplant studies involving in vitro manipulation of hematopoietic pro. PMID- 10459538 TI - Significant progress in porcine islet mass isolation utilizing liberase HI for enzymatic low-temperature pancreas digestion. AB - BACKGROUND: Frequent success in human islet isolation is prevented by the large variability of scarce organ donors; this favors the future utilization of pigs as donors for clinical islet xenotransplantation. Porcine-specific difficulties of islet isolation are attributed to the intrinsic fragility of islets during pancreas digestion. METHODS: To preserve islet integrity during efficient pancreas dissociation, porcine pancreata (n=48) were distended after cold storage with cold University of Wisconsin solution containing Liberase HI and digested at 24-28 degrees C using digestion-filtration. Pancreata distended with University of Wisconsin solution containing well-proven crude collagenase and digested at 32 34 degrees C served as controls (n=46). Monolayer Ficolldiatrizoate gradient purification was performed in a Cobe 2991. RESULTS: Purified yield of islet equivalents per pancreas (mean+/-SEM) was almost doubled by Liberase HI compared with crude collagenase (526,480+/-46,560 vs. 270,270+/-19,420; P < 0.0001) and also significantly increased comparing islet equivalents per gram of pancreas (4,210+/-320 vs. 2,640+/-245; P=0.0004). Islet integrity was better preserved during Liberase HI digestion compared with crude collagenase digestion as indicated by isolation index (2.1+/-0.1 vs. 1.4+/-0.1; P<0.0001). Purity, viability, and in vitro function of islets did not differ between experimental groups. Preserved in vivo function of islets isolated by Liberase HI was demonstrated after subcapsular transplantation into 16 diabetic nude rats. CONCLUSIONS: If the problems related to xenograft rejection and xenosis could be solved, low-temperature digestion of porcine pancreata using Liberase HI could serve as an essential prerequisite for successful 1:1 xenotransplantation of pig islets into type 1 diabetic human recipients. PMID- 10459537 TI - Improved preservation solutions for organ storage: a dynamic study of hepatic metabolism. AB - BACKGROUND: Organ cold storage times may be extended by modifications to organ preservation solutions. METHODS: Three preservation solutions were investigated for their ability to maintain viable hepatic bioenergetics in stored pig livers: modified University of Wisconsin (mUW); mUW+adenosine (1.34 g/L), and mUW+ iloprost (10(-8)mol/L), a prostacyclin analogue. Using human liver retrieval and storage techniques, pig livers were stored on ice for either 2 or 16 hr, after which phosphorus-31 spectra were collected every 2 min during the period of cold ischemia and hypothermic reperfusion (HtR). During HtR, metabolite concentration changes associated with phosphomonoesters, inorganic phosphate, gamma-nucleotide triphosphate (NTP), and beta-NTP were measured for all solutions. RESULTS: After a 2-hr storage, beta-NTP regeneration in mUW+iloprost produced +57.7% (P<0.01) more beta-NTP, at a faster initial rate of +66.3% (P<0.001), compared with mUW, and mUW+adenosine regenerated +35.6% (P<0.05) more beta-NTP, compared with mUW. Storage for 16 hr did not slow the rates of regeneration, and the total NTP produced during the course of the experiment remained unchanged for the respective preservation solutions. Cessation of HtR invoked a net accumulation of nucleotide diphosphate, indicating differential kinetics of adenine nucleotide hydrolysis. CONCLUSION: This large animal model study suggests significant improvements to human organ preservation solutions using prostacyclin analogues and adenosine with respect to hepatic bioenergetics. PMID- 10459539 TI - Biliary secretion of extracorporeal porcine livers with single and dual vessel perfusion. AB - BACKGROUND: Hepatic support systems that provide detoxification without biliary secretion (i.e., isolated hepatocyte systems) are sufficient to improve encephalopathy and bridge patients to transplantation. However, biliary secretion may be critical when hepatic support attempts to restore function and regeneration of the host liver. The purpose of these studies was to optimize the support liver secretory response to bile acid by either single-vessel (portal vein; PV) or dual-vessel (hepatic artery [HA] + PV) perfusions during extracorporeal porcine liver perfusion. METHODS: Extracorporeal porcine liver perfusion of anesthetized pigs was developed using support porcine livers perfused through the PV (n=4) alone and through the HA + PV (n=4) via a venovenous circuit. Support livers were provoked with taurocholate (TC) to enhance bile aqueous and hydrophobic outputs. RESULTS: After cold preservation and reperfusion, both PV and HA + PV livers had initial 1-hr bile aqueous outputs < 15% of in vivo flow, with cholesterol (C) and phospholipid (PC) outputs <25% of in vivo flow. Bile flow was significantly greater for recovered HA + PV livers (3.0+/-0.01 ml/15 min) than PV livers (1.9+/-0.01 ml/15 min). Despite this, PC output was significantly greater for PV than HA + PV livers. The C/PC ratio of PV livers was twice that of HA + PV livers. TC infusion (48 micromol/kg/15 min) of HA + PV livers demonstrated significantly greater increments in bile flow, PC output, and C output than PV livers. CONCLUSION: In the unstimulated state, porcine support livers with dual-vessel perfusion generated greater aqueous and C outputs despite diminished PC output than in those with single-vessel perfusion. TC stimulation increased bile flow, PC output, and C output in dual-perfused livers more than in PV livers. HA + PV perfusion of support livers is the preferred technique for removing hydrophobic compounds that require PC transport for excretion or exist in the aqueous phase. PMID- 10459540 TI - A partial conditioning approach to achieve mixed chimerism in the rat: depletion of host natural killer cells significantly reduces the amount of total body irradiation required for engraftment. AB - BACKGROUND: Mixed allogeneic bone marrow chimerism induces tolerance to solid organ grafts. Although we previously reported that partially ablative conditioning with 700 cGy of total body irradiation (TBI) is sufficient to allow for bone marrow engraftment in mice, we determined that a minimum of 1000 cGy was required in the rat. Because T cells and NK cells are critical in bone marrow graft rejection, our purpose was to examine whether targeting of radioresistant NK cells and/or T cells in the recipient hematopoietic microenvironment would reduce the TBI dose required for engraftment of allogeneic rat bone marrow. METHODS: Wistar Furth rats received either anti-NK3.2.3 monoclonal antibodies on days -3 and -2, anti-lymphocyte serum on day -5, a combination of both or no pretreatment. TBI was performed on day 0 and rats were reconstituted with 100x10(6) T cell-depleted bone marrow cells from ACI donors. RESULTS: Engraftment of T cell-depleted rat bone marrow was readily achieved in animals conditioned with 1000 cGy TBI alone (12/12) and the level of donor chimerism averaged 89%. At 900 cGy TBI alone only one of eight recipients engrafted. In striking contrast, 11 of 12 animals pretreated with anti-NK monoclonal antibodies and irradiated with 900 cGy showed donor chimerism at a mean level of 41%. No further enhancement of bone marrow engraftment could be achieved when recipients were pretreated with antilymphocyte serum alone or antilymphocyte serum plus anti-NK monoclonal antibodies. Mixed allogeneic chimeras exhibited stable multilineage chimerism and donor-specific tolerance to subsequent cardiac allografts. CONCLUSION: Specific targeting of radioresistant host NK cells allows for a significant reduction of the TBI dose required for allogeneic bone marrow engraftment. PMID- 10459541 TI - Transjugular intrahepatic portosystemic shunt after adult liver transplantation: experience in eight patients. AB - BACKGROUND: Transjugular intrahepatic portosystemic shunting (TIPS) has become an effective treatment for the complications of portal hypertension. We assessed the feasibility and outcome of TIPS in liver transplant recipients. METHODS: During the period from December 1992 to January 1998, eight adults presenting recurrent hepatitis C virus (five patients) and hepatitis B virus (one patient) infection, veno-occlusive disease (one patient), and secondary biliary cirrhosis (one patient) had TIPS because of refractory ascites (five patients), bleeding esophageal varices (one patient), refractory hepatic hydrothorax (one patient), retransplantation (two patients), and redo-biliary surgery (one patient). RESULTS: In two patients, the procedure was difficult due to cavo-caval implantation. Ascites, hydrothorax, and variceal bleeding were controlled in all patients. Moderate to severe encephalopathy developed in four patients; two patients had worsening of their existing encephalopathy. Three of five patients treated with cyclosporine needed a drastic dose reduction due to the development of severe side effects. No long-term survivor developed shunt stenosis or occlusion. Two patients did moderately well at 6 and 14 months, respectively; the former died due to chronic rejection while waiting for a retransplantation. Three did well at 14, 36, and 28 months, respectively; the latter patient died of liver failure 32 months after TIPS. One jaundiced patient died after 1.5 months due to necrotic pancreatitis. Two patients died after 4 and 8.5 months, respectively, due to liver failure; the latter was doing well until 7 months after TIPS. CONCLUSIONS: TIPS is feasible in transplant recipients in cases of decompensated allograft cirrhosis, of allograft veno-occlusive disease or when retransplantation or redo-biliary surgery are scheduled in the presence of portal hypertension. At transplantation, the surgeon should keep in mind the eventuality of a later TIPS procedure. Close immunosuppression monitoring is warranted because modified metabolization of cyclosporine (and probably tacrolimus) may cause serious side effects. PMID- 10459542 TI - Renal neoplasias in patients receiving dialysis and renal transplantation: clinico-pathological features and p53 gene mutations. AB - BACKGROUND: In Japan, the relative risk for renal cell carcinoma (RCC) in renal transplants was about 80-fold higher than that in the general population. Depressed immune surveillance due to the use of immunosuppressive agents was considered to cause cancer. Before renal transplantation, a vast majority of patients received hemodialysis, a known causative factor for acquired cystic disease of kidney (ACDK). Because ACDK is also considered to predispose to RCC, at least two risk factors for cancer accumulate in renal transplants. METHODS: In our study, clinicopathological features together with p53 gene mutations were analyzed in 218 patients with RCC: 22 received dialysis followed by renal transplantation, 39 received dialysis alone, and 157 sporadic RCC. P53 mutations were analyzed on DNA extracted from paraffin-embedded specimens with use of single strand conformation polymorphism, followed by direct sequencing. RESULTS: RCC in transplants shared several clinicopathological features with those in dialysis patients, which included small size and multiplicity of tumor, relatively high frequency of presence of ACDK, and papillary type of RCC. p53 gene mutations were infrequent in RCC of any clinical setting. CONCLUSIONS: Atrophic kidney at the end-stage of renal failure and under dialysis have lesions of ACDK that might predispose to RCC in dialysis and transplant patients. PMID- 10459543 TI - Mycophenolate mofetil in renal transplantation: 3-year results from the placebo controlled trial. European Mycophenolate Mofetil Cooperative Study Group. AB - BACKGROUND: The European double-blind, placebo(PLA) controlled study of mycophenolate mofetil (MMF) for prevention of acute renal allograft rejection showed that MMF 2 and 3 g when added to a standard double-drug regimen of cyclosporine and corticosteroids significantly reduced the incidence of acute rejection/treatment failure at 6 months. Our study presents 3-year data for patient and graft survival, and safety in the MMF-treated patients. METHODS: The trial included 491 patients who were randomly assigned to receive PLA (n=166), MMF 2g (n=165), or MMF 3 g (n=160). Patients in the PLA group discontinued taking their PLA medication at 1 year posttransplantation; subsequently, they were followed-up only regarding patient and graft survival and the occurrence of malignancies. RESULTS: The 3-year patient survival was 88.9, 92.7, and 91.8% in the PLA, MMF 2 g, and MMF 3 g groups, respectively. The 3-year graft survival (including death as a cause of graft loss) was 78, 84.8, and 81.2%, respectively. Acute allograft rejection was a principal cause of graft loss in all groups (PLA, 10.8%; MMF 2 g, 4.6%; MMF 3 g, 6.3%). Differences in 3-year graft loss rates (excluding death) and 95% confidence intervals for intent-to-treat comparisons of PLA versus MMF 2 g and 3 g, respectively, were 7.3% (1.1, 14.2) and 3.2% (-3.8, 10.1). This leads to a relative risk of graft loss of 0.55 in the MMF 2 g arm compared with the PLA arm. Acute allograft rejection had a major impact on graft loss at 3 years; 31.5% of patients with biopsy-proven acute rejection within 6 months of transplantation lost their graft by the end of 3 years. In contrast, only 6.6% who had no early acute rejection lost their graft by the end of the 3 year study period. Diarrhea, anemia, and leukopenia were the most common clinically relevant adverse events, occurring predominantly in the MMF 3 g group. Only one patient (MMF 3 g) developed cytomegalovirus tissue-invasive disease after the first year posttransplant. Over the 3-year posttransplant period, 12 patients developed malignancies (5 in the PLA group, 3 in the MMF 2 g group, and 4 in the MMF 3 g group). CONCLUSIONS: At 3 years posttransplantation, MMF was associated with 7.6% reduction in the incidence of graft loss (excluding death). These data indicate that MMF treatment not only results in a reduction of the incidence of acute rejections but also leads to reduction of late allograft loss. PMID- 10459544 TI - Islet cell damage associated with tacrolimus and cyclosporine: morphological features in pancreas allograft biopsies and clinical correlation. AB - BACKGROUND: The introduction of the potent immunosuppressive drugs tacrolimus (FK) and cyclosporine (CSA) has markedly improved the outcome of solid organ transplantation. However, these drugs can cause posttransplantation diabetes mellitus. Abnormalities in the glucose metabolism are of particular significance in pancreas transplantation. METHODS: We studied 26 pancreas allograft biopsies, performed 1-8 months posttransplantation, from 20 simultaneous kidney-pancreas transplant recipients, randomized to receive either FK or CSA. The biopsies were studied by light microscopy, immunoperoxidase stains for insulin and glucagon, in situ DNA-end labeling for detection of apoptosis, and electron microscopy. The islet morphology was correlated with the mean and peak levels of CSA and FK in serum, with corticosteroid administration and with glycemia. RESULTS: On light microscopy cytoplasmic swelling, vacuolization, apoptosis, and abnormal immunostaining for insulin were seen in biopsies from patients receiving either FK or CSA. The islet cell damage was more frequent and severe in the group receiving FK than in the group receiving CSA (10/13 and 5/13, respectively) but the differences were not statistically significant. Significant correlation was seen between the presence of islet cell damage and serum levels of CSA or FK during the 15 days previous to the biopsy, as well as with the peak level of FK. Toxic levels of CSA or FK and administration of pulse steroids were associated with hyperglycemia when these occurred concurrently (P=0.005). Toxic levels of CSA or FK by themselves were associated with hyperglycemia in a minority of cases (8 and 26%, respectively). Electron microscopy showed cytoplasmic swelling and vacuolization, and marked decrease or absence of dense-core secretory granules in beta cells; the changes were more pronounced in patients on FK. Serial biopsies from two hyperglycemic patients receiving FK and evidence of islet cell damage demonstrated reversibility of the damage when FK was discontinued. CONCLUSIONS: The structural damage to beta cells demonstrated in this study is similar to morphological and functional abnormalities previously described in experimental animal models and can at least partially account for the glucose metabolism abnormalities seen in patients receiving these drugs. Toxic levels of CSA or FK and higher steroid doses potentiate each others' diabetogenic effects. PMID- 10459545 TI - Positive and negative selection of alphabetaTCR+ T cells in thymectomized adult radiation bone marrow chimeras. AB - BACKGROUND: The mature T-cell repertoire is characterized by the negative selection of potentially autoreactive T cells and the positive selection of T cells restricted to antigen-recognition in the context of self-MHC molecules. It is currently believed that the thymus is critical for these selection events. Although alpha(beta)T cell receptor (TCR)+ T cells have been reported in thymectomized recipients, whether this represents clonal expansion of residual T cells or de novo generation of new T cells in the absence of a thymus has not been definitively evaluated. METHODS: In the current study, development of the T cell repertoire was evaluated in adult radiation bone marrow chimeras prepared after complete surgical thymectomy. RESULTS: CD4+ and CD8+ T cells were present and exhibited donor-specific TCR-Vbeta expression and self-tolerance, indicative of negative selection. Positive selection was confirmed with the demonstration of host MHC restriction and the presence of donor-derived CD8+ T cells after the transplantation of marrow from Class I deficient donors into normal recipients. CONCLUSIONS: These data provide evidence, for the first time, that the development of a functional T-cell repertoire can occur in adult recipients without the thymic microenvironment. PMID- 10459546 TI - Mycophenolic acid increases apoptosis, lysosomes and lipid droplets in human lymphoid and monocytic cell lines. AB - BACKGROUND: Mycophenolic acid (MPA), a selective inhibitor of inosine monophosphate dehydrogenase, is the active agent of the immunosuppressive drug, mycophenolate mofetil (MMF). Previous studies have shown that MPA inhibits DNA synthesis in T and B lymphocytes by blocking de novo guanosine synthesis, and that MPA induces monocyte differentiation. MMF is being used for prevention of organ graft rejection and has also shown efficacy in rheumatoid arthritis trials. This study was designed to determine if apoptosis also plays a role in the immunosuppressive and anti-inflammatory effects of MMF. METHODS: Cultured human T lymphocytic (MOLT-4) and monocytic (THP-1 and U937) cell lines were treated with MPA. Apoptosis, cell viability, DNA content, lipid content, cell volume, and lysosomes were measured by a variety of microscopic, flow cytometric, and biochemical techniques. RESULTS: MPA inhibits proliferation, arrests cell cycle in S phase, and increases apoptosis in all three cell lines. Exogenous guanosine added within 24 hr of MPA treatment, but not later, partially reversed MPA induced apoptosis in MOLT-4 cells. MPA increased lipid droplets in all three cell lines and increased both cell volumes and numbers of lysosomes in the monocytic cell lines. In both monocytic cell lines, MPA also reduced the number of nuclei containing nucleoli and greatly increased neutral lipids, primarily triacylglycerols, suggesting that these cells were differentiating. CONCLUSIONS: Increased apoptosis and terminal differentiation of both lymphocytes and monocytes may promote the antiproliferative, immunosuppressive, and anti inflammatory effects of MMF seen clinically in transplantation and rheumatoid arthritis. PMID- 10459547 TI - Mechanism of acquired thymic tolerance induced by a single major histocompatibility complex class I peptide with the dominant epitope: differential analysis of regulatory cytokines in the lymphoid and intragraft compartments. AB - BACKGROUND: We have recently shown that intrathymic injection of a combination of immunogenic WAG-derived or Wistar-Furth (WF) (RT1.Au) major histocompatibility complex class I peptides induces acquired systemic tolerance to cardiac and islet allografts in the WF-to-ACI rat combination and therefore hypothesized that identification of the class I peptide dominance may play an important role in the induction of antigen (Ag)-specific tolerance. This study defined the peptide with the dominant epitope among the seven synthetic RT1.Au peptides and analyzed the immunoregulatory cytokines within the lymphoid and intragraft compartments associated with acquired thymic tolerance. METHODS: ACI recipients were pretreated with intrathymic (IT) injection of 300 microg of the individual seven RT1.Au peptides 7 days before WF or Lewis cardiac transplantation. Cytokine profile of mixed lymphocyte reaction supernatants of T cells obtained from the thymus, mesenteric lymph nodes, spleen, peripheral blood leukocytes, and graft infiltrating cells after donor (WF) or third-party (Lewis) Ag stimulation were measured by enzyme-linked immunosorbent assay, whereas cytokine gene expression was determined by reverse transcription-polymerase chain reaction. RESULTS: Only IT injection of peptide 5 (93-109) among the seven RT1.Au peptides induced donor spe cific tolerance to cardiac allografts in the WF-to-ACI rat combination. In addition, intravenous injection of peptide 5 did not prolong WF graft survival in ACI recipients. Analysis of cytokine production by the tolerant recipients showed significant Ag-specific reduction in the production of interleukin (IL)-2 and interferon-gamma (IFN-gamma) in the thymus, mesenteric lymph nodes, spleen, and peripheral blood leukocytes, which was not associated with a concomitant Ag specific increase in IL-4 and IL-10 production. Measurement of cytokine mRNA expression confirmed undetectable PMID- 10459548 TI - CD40 ligation induced phenotypic and functional expression of CD80 by human cardiac microvascular endothelial cells. AB - BACKGROUND: CD40/CD40L (gp39) interactions are known to play a central role in the function of the immune system (1). CD40 is constitutively expressed on professional antigen presenting cells, such as macrophages and dendritic cells, as well as at low levels on other cell lineages, including human umbilical vein endothelial cells (HUVECs). On antigen-presenting cells, ligation of CD40 causes expression of the costimulatory molecule CD80 (B7-1). Similar ligation of CD40 on HUVECs, however, leads to up-regulation of the adhesion molecules VCAM-1, ICAM-1, and E-selectin, but not CD80. METHODS: In efforts to provide evidence that microvascular endothelial cells (MECs) are distinct from HUVECs and that the distinguishing features play a role in allograft rejection, MEC cultures were prepared from the explanted hearts of human heart transplant recipients and primary cell lines were established. These MECs were induced to express higher levels of CD40 with interferon-gamma pretreatment, co-cultured with CD40L transfected HeLa cells, and fluorescence-activated cell sorter-assisted phenotypic studies, in addition to functional allogeneic mixed lymphocyte reaction and accessory-cell dependent mitogen induced proliferation assays were performed. RESULTS: CD40-CD40L interactions induced the expression of the adhesion molecules VCAM-1 and E-selectin and the costimulatory molecule CD80 but not CD86 (B7-2) on the MECs. The expressed CD80 proved functional in both allo MLR assays and accessory-cell dependent mitogen proliferation assays. CONCLUSIONS: MECs are distinct from HUVECs by their potential to express VCAM-1 after interferon-gamma pretreatment and CD80 after CD40 ligation, properties which enable this cell lineage to play a central role in initiating and maintaining allograft rejection in human cardiac transplants. PMID- 10459549 TI - Protection of sinusoidal endothelial cells against storage/reperfusion injury by prostaglandin E2 derived from Kupffer cells. AB - BACKGROUND: In clinical liver transplants, grafts are frequently exposed to endotoxin (lipopolysaccharide, LPS) before harvest and may be predisposed to dysfunction. Because graft failure is linked to sinusoidal endothelial cell injury after storage/reperfusion, we investigated the effect of donor exposure to LPS on graft survival in relation to sinusoidal endothelial cell injury after storage/reperfusion in rats. METHODS: Rats were injected with 0.5 mg/kg LPS. In some rats, 20 mg/kg GdCl3 or 5 mg/kg indomethacin was injected before LPS to ablate Kupffer cells and inhibit prostaglandin (PG) synthesis, respectively. Other rats were injected with 100 microg/kg dimethyl PGE2, a stable PGE2 analog. Rat livers were harvested, stored in cold UW solution and transplanted to non treated rats for determination of survival and liver injury in recipients. Otherwise, after cold storage, the livers were reperfused briefly with physiological buffer containing trypan blue for determination of sinusoidal endothelial cell injury by counting trypan blue-positive nuclei in histological sections. RESULTS: Donor treatment with LPS increased hepatic PGE2 production before storage and decreased recipient survival, but paradoxically decreased killing of sinusoidal endothelial cells after storage and reperfusion. Pretreatment of donors with GdCl3 or indomethacin prevented the protective preconditioning of sinusoidal endothelial cells by LPS, whereas pretreatment with dimethyl PGE2 protected sinusoidal endothelial cells to the same extent as LPS. Unlike LPS, however, PGE2 attenuated graft injury after liver transplants. CONCLUSION: PGE2 derived from LPS-stimulated Kupffer cells protects sinusoidal endothelial cells against storage/reperfusion injury. Unlike LPS, PGE2 improves graft function after liver transplants. Thus, donor preconditioning with PGE2 may be beneficial in liver transplants. PMID- 10459550 TI - Endovascular treatment of hepatic venous outflow obstruction after piggyback technique liver transplantation. AB - BACKGROUND: The piggyback technique of orthotopic liver transplantation is an attractive alternative that preserves the recipient inferior vena cava and allows uninterrupted venous blood return during the anhepatic phase. As with other transplantation techniques, the vascular anastomoses required by the piggyback technique can develop strictures. METHODS: Review of records of 264 piggyback transplantations revealed two cases of delayed-onset hepatic venous obstruction from anastomotic strictures. Both patients also had symptoms of inferior vena cava obstruction, with azotemia and lower extremity edema. Both patients were treated percutaneously with balloon-expandable stents. RESULTS: Rapid, dramatic resolution of symptoms was achieved in both patients. Patients remain completely asymptomatic at 39 and 3 months of follow-up. CONCLUSIONS: Hepatic venous anastomotic strictures in recipients of piggyback technique transplants are a very uncommon complication. They may be easily and effectively treated by minimally invasive endovascular intervention. PMID- 10459551 TI - Complication after laparoscopic donor nephrectomy: a case report and review. AB - BACKGROUND: Laparoscopic procedures are gaining acceptance in the treatment of benign and some malignant urologic disorders. Recently, laparoscopic techniques have been applied to transplant surgery and touted as a safe alternative to traditional open techniques. METHODS: We present a patient who developed a complication from laparoscopic donor nephrectomy that required open corrective surgery. RESULTS: A 25-year-old man underwent laparoscopic donor nephrectomy at a large medical center familiar with the operation. There were no operative or early postoperative complications. Within 6 weeks of the operation, the patient developed signs and symptoms of partial small bowel obstruction. Further evaluation revealed an internal hernia in the retroperitoneum at the site of the nephrectomy. This required a second operation to reduce the hernia and close the defect. CONCLUSION: Laparoscopic donor nephrectomy remains an evolving technique that has not stood the test of time. Larger series will eventually reveal whether this is the procedure of choice as compared to traditional open donor nephrectomy. PMID- 10459552 TI - Delayed graft function and renal allograft outcome. PMID- 10459553 TI - Apolipoprotein E genotype and exercise training-induced increases in plasma high density lipoprotein (HDL)- and HDL2-cholesterol levels in overweight men. AB - We determined if the apolipoprotein E (APO E) genotype affects the exercise training-induced increase in plasma high-density lipoprotein cholesterol (HDL-C) and HDL2-C. Sedentary overweight men on an American Heart Association (AHA) step I diet had plasma lipoprotein-lipids measured before and after 9 months of endurance exercise training. APO E2 (n = 6), E3 (n = 33), and E4 (n = 12) groups were similar at baseline in terms of age, body weight and composition, and plasma lipoprotein-lipid profiles. APO E2 men had a larger increase in plasma HDL-C and HDL2-C with exercise training than APO E3 and E4 men (HDL-C, 8 +/- 4 v 3 +/- 1 v 2 +/- 1 mg/dL; HDL2-C, 5 +/- 3 v 1 +/- 1 v -1 +/- 1 mg/dL; mean +/- SE, all P < .01). After adjusting for body weight changes, the increases in plasma HDL-C and HDL2-C remained greater in APO E2 versus E3 and E4 men (all P < .03). These results indicate that APO E2 men may have greater plasma HDL-C and HDL2-C increases with endurance exercise training. PMID- 10459554 TI - Higher body fat aggravates toxin-induced infectious episodes. AB - Animal models using rabbits were developed to accumulate a variable body fat mass (FM) in two groups of animals while the fat-free mass (FFM), eg, total body protein, was maintained essentially similar between the groups. Thiobarbituric acid-reactive substances (TBARS) were measured as an index of lipid peroxidation and were found to be higher in the whole-body tissues of animals with a higher FM. Bacterial toxin (lipopolysaccharide [LPS]) also induced higher lipid peroxidation in animals with a higher FM, with a concomitant incidence of bloody mucous diarrhea. To our knowledge, this is the first evidence to show the effect of body FM to aggravate toxin-induced infections leading to diarrhea. The overall results suggest further investigations to explore the possible role of body fat in infectious diseases in humans. PMID- 10459555 TI - Adenosine deaminase and body mass index in non-insulin-dependent diabetes mellitus. AB - We studied 273 subjects with non-insulin-dependent diabetes mellitus (NIDDM) from the population of Penne, Italy. A low proportion of the adenosine deaminase (ADA)*2 allele is observed in NIDDM subjects with a body mass index (BMI) of 25 kg/m2 or less. On the contrary, a high proportion of this allele is observed in NIDDM patients with a BMI higher than 34 kg/m2. In the intermediate BMI class, the proportion of ADA*2 alleles does not differ significantly from that of normal subjects from the same population. No significant effect on the relation between ADA and BMI has been observed for the following variables: sex, age at the time of study, age at onset, therapy with insulin, and dyslipidemia. A borderline effect has been observed for the duration of disease. Several lines of experimental evidence suggest that an excess of adenosine A1 receptor activity may contribute to adiposity in NIDDM. ADA is a polymorphic enzyme that irreversibly deaminates adenosine to inosine, contributing to the regulation of intracellular and extracellular concentrations of adenosine. Since the activity of genotypes carrying the ADA*2 allele is lower than that of the more common genotype ADA*1/*1, genetic variability of the enzyme could contribute to degree of obesity in NIDDM. Our data also support attempts to ameliorate the metabolic control of diabetes through pharmacological modulation of adenosine receptors. PMID- 10459556 TI - Multivariate identification of metabolic features in inflammatory bowel disease. AB - Weight loss and malnutrition are commonly reported in inflammatory bowel disease (IBD), but differences between Crohn's disease (CD) and ulcerative colitis (UC) patients have rarely been pointed out. In this regard, a sample of 102 consecutive patients with a diagnosis of either CD (n = 63, 33 males) or UC (n = 39, 25 males) based on previously reported clinical, morphologic, and histopathologic criteria were studied. Twenty-six anthropometric and metabolic variables were measured upon admission. Body composition was assessed by both anthropometry and bioimpedance measurements, and energy expenditure and substrate oxidation were assessed by indirect calorimetry. The data were subjected to principal-component analysis and to factor rotation to derive a set of a few basic independent descriptors of the metabolic features of each subject. Six descriptors were found to be responsible for greater than 86% of the total sample variability and to associate very well with mutually disjoint subsets of the original variables. The six summarizing factors are listed in order of decreasing percentage of explained variation (size 41.8%, fatness 17.9%, fuel 12.2%, shape 5.4%, energy 5.2%, and steroid 3.9%). CD and UC patients differed significantly with respect to fatness (CD lower, P = .004) and carbohydrate (CHO) fuel preference (CD lower, P = .030). Hence, CD patients showed a reduced fat mass (FM) compared with UC patients, and from a metabolic point of view, too, CD and UC are not superimposable. In fact, the lower CHO oxidation (CHOox) rate and consequent preferential lipid utilization found in CD patients may be taken into account as a contributing cause of lipid tissue wasting and in planning therapeutic enteral regimens. PMID- 10459557 TI - Effect of vitamin E and N-acetylcysteine on phosphatidylserine externalization and induction of coagulation by high-glucose-treated human erythrocytes. AB - This study examines the effect of high glucose levels on the markers of oxidative stress, phosphatidylserine (PS) externalization, and induction of coagulation by high-glucose-treated red blood cells (RBCs). Washed normal RBCs were suspended to 15% hematocrit in phosphate-buffered saline and incubated with different concentrations of glucose for 24 hours in a shaking water bath at 37 degrees C. This treatment caused depletion of vitamin E and accumulation of vitamin E quinone and malondialdehyde ([MDA] an end product of lipid peroxidation), externalization of PS in the membrane bilayer, and induction of coagulation by RBCs. Pretreatment of RBCs with N-acetylcysteine (NAC) and vitamin E reduced membrane lipid peroxidation, PS externalization, and the tendency of high-glucose treated RBCs to clot plasma. This study provides further evidence for the increased oxidative stress in RBCs exposed to high glucose levels. In addition, it suggests a role for membrane lipid peroxidation in the PS externalization in the membrane bilayer and in the induction of clotting by RBCs exposed to hyperglycemia. It also suggests that certain antioxidants can decrease cellular damage and restore certain cellular functions in diabetes. PMID- 10459558 TI - Plasma plasminogen activator inhibitor-I is associated with plasma leptin irrespective of body mass index, body fat mass, and plasma insulin and metabolic parameters in premenopausal women. AB - Leptin, the satiety hormone expressed almost exclusively in adipose tissue, is a marker of body fat accumulation in humans. Recent studies have shown that plasminogen activator inhibitor-1 (PAI-1), a prothrombotic factor associated with atherosclerosis complications, is also produced in adipose tissue. The objective of the present study was to determine whether PAI-1 antigen plasma concentrations are associated with leptin plasma levels or the body fat mass (FM) independently of the variables known to influence PAI-1 production. Sixty-one nondiabetic women aged 18 to 45 years with a wide range of values for the body mass index ([BMI] 18.1 to 37.7 kg/m2) were evaluated for (1) body FM and fasting plasma levels of (2) PAI-1 antigen, (3) PAI-1 activity, (4) leptin, (5) insulin, (6) blood glucose, and (7) lipids (cholesterol, high-density lipoprotein [HDL]-cholesterol, and triglycerides [TG]). Body FM and fat-free mass (FFM) were estimated during fasting conditions by the bioimpedance analysis (BIA) method using a tetrapolar device. Body fat distribution was evaluated by the waist circumference and the waist to hip ratio (WHR). FM was directly associated with both PAI-1 antigen (r = .585, P < .001) and PAI-1 activity (r = .339, P < .001). Seemingly, leptin was positively related to both PAI-1 antigen (r = .630, P < .001) and PAI-1 activity (r = .497, P < .001). Moreover, both PAI-I antigen and PAI-1 activity were directly correlated with FFM (r = .285, P < .05, and r = .336, P < .01, respectively), BMI (r = .594, P < .001, and r = .458, P < .001, respectively), and WHR (r = .510, P < .001, and r = .391, P < .005, respectively). Insulin was directly related to PAI-1 antigen (r = .540, P < .001), PAI-1 activity (r = .259, P < .05), leptin (r = .447, P < .001), and FM (r = .435, P < .001). The association between PAI-1 antigen (dependent variable) and leptin or FM was tested by a stepwise regression model simultaneously including leptin, FM, BMI, WHR, age, FFM, and fasting insulin, blood glucose, TG, cholesterol, and HDL cholesterol as independent variables. PAI-1 antigen maintained a significant positive independent relationship only with leptin (t = 2.923, P < .01), insulin (t = 3.489, P < .001), and fasting blood glucose (t = 2.092, P < .05), and a negative independent relationship with HDL-cholesterol (t = -2.634, P < .05). In conclusion, the strong relationship between PAI-1 antigen and leptin irrespective of other variables known to influence these factors seems to indicate that leptin per se may potentially increase PAI-1 plasma concentrations in obese subjects. PMID- 10459559 TI - Dose-response relationship of insulin to glucose fluxes in the awake and unrestrained mouse. AB - The purpose of the study was to use the hyperinsulinemic-euglycemic clamp technique to generate insulin dose-response curves for insulin suppression of endogenous glucose output (EGO) and stimulation of the glucose disposal rate (GDR) in conscious unstressed mice. Five groups of male ICR (Institute for Cancer Research) mice were studied (N = 43). The animals underwent surgery for implantation of a jugular vein catheter 2 to 3 days before the clamp and were fasted 6 hours before the study. Each group was clamped at a different insulin infusion rate of 0, 2.5, 10, or 20 mU/kg/min. 3H-3-glucose was infused for measurement of the glucose turnover rate (rate of appearance [Ra]). Blood samples were collected by milking a severed tail-tip. EGO was calculated as the difference between the Ra and glucose infusion rate (GIR), and the glucose clearance rate (GCR) as the GDR divided by the plasma glucose concentration. From the curves generated, half-maximal EGO and GCR were obtained at a plasma insulin concentration of 20 to 30 microU/mL, which was achieved at an insulin infusion rate of about 4 to 5 mU/kg/min. Maximal suppression of EGO and stimulation of the GCR occurred at an insulin infusion rate of 10 mU/kg/min. The establishment of normative curves for insulin-stimulated glucose metabolism in conscious mice facilitates the evaluation of glucose metabolism in a variety of mouse models of insulin resistance. PMID- 10459560 TI - Effect of exercise on postprandial insulin responses in Mexican American and non Hispanic women. AB - Postprandial insulin responses (integrated area under the curve) to an oral glucose load after a period of aerobic exercise and no exercise (control) were compared in sedentary normoglycemic Mexican American and non-Hispanic women pair matched (n = 9) on total body fat mass (21.8 +/- 3.5 kg). The age (27.4 +/- 3.0 years), body mass index (BMI) (23.6 +/- 1.4 kg/m2), waist to hip ratio (WHR) (0.85 +/- .02), waist circumference (83.5 +/- 4.5 cm), lean mass (36.2 +/- 1.5 kg), and maximal O2 consumption ([VO2 max] 32.9 +/- 1.6 mL x kg(-1) x min(-1)) were similar, although the centrality index (subscapular/triceps skinfolds) was significantly greater in Mexican Americans (0.88 +/- 0.06 v 0.70 +/- 0.05, P < .01). Exercise (treadmill walking for 50 minutes at 70% VO2 max) and control trials were performed 4 weeks apart and 5 to 12 days after the onset of menstruation. A 75-g oral glucose load was administered 15 hours after the completion of each trial, with the subjects 12 hours postprandial. Blood samples were drawn prior to glucose ingestion (fasting, 0 minutes) and at minutes 15, 30, 60, 90, 120, and 150 postingestion. The postprandial insulin response was calculated using a trapezoidal method. In Mexican Americans, significant (P < .02) reductions in the postprandial insulin response (exercise v control, 6.5 +/- 1.0 v 8.5 +/- 1.4 pmol/L x min x 10(4)) and fasting insulin (exercise v control, 77.4 +/- 7.0 v 88.5 +/- 10.3 pmol/L) occurred after exercise compared with the control condition. In non-Hispanics, neither the postprandial insulin response (exercise v control, 7.2 +/- 1.0 v 6.2 +/- 0.9 pmol/L x min x 10(4)) nor fasting insulin (exercise v control, 77.0 +/- 8.2 v 82.9 +/- 8.9 pmol/L) were significantly different between trials. The postprandial insulin response in the control trial was significantly correlated with the change in the insulin response (control minus exercise) in the 18 women (r = .56, P = .01). No trial or group differences were found for postprandial glucose and C-peptide responses. Mexican American women have a high risk of developing type 2 diabetes, and aerobic exercise may be valuable in the prevention or delay of onset of diabetes by reducing peripheral insulin resistance. PMID- 10459561 TI - Insulin sensitivity, glucose effectiveness, and insulin secretion in nondiabetic offspring of patients with non-insulin-dependent diabetes mellitus: a cross sectional study. AB - To evaluate the factors that determine the worsening of intravenous glucose tolerance in subjects at high risk for developing non-insulin-dependent diabetes mellitus (NIDDM), 15 glucose-tolerant offspring of NIDDM patients and 21 control subjects were studied. Each subject underwent a frequently sampled intravenous glucose tolerance (FSIGT) test. The intravenous glucose tolerance index (K(G) index) was calculated between minutes 10 and 40 of a FSIGT test. Insulin sensitivity (S(I)), glucose effectiveness at zero insulin (GEZI), and first- and second-phase insulin responsiveness (phi1 and phi2) were estimated using glucose and insulin kinetic minimal models. The acute insulin response to glucose (AIRg) was calculated as the area under the insulin curve above the basal level between 0 and 10 minutes, and the suprabasal insulin effect was determined by the product of S(I) times AIRg. Offspring had a lower S(I) than control subjects (14.1 +/- 7.5 v 9.25 +/- 4.20 x 10(-5) x min(-1)(pmol x L(-1))(-1), P < .01), and their AIRg was similar (3,284 +/- 2,280 v 3,105 +/- 1,499 pmol x L(-1), NS). Sample division according to the median K(G) value showed that control subjects with low tolerance had a lower AIRg (4,417 +/- 2,531 v 2,043 +/- 1,068 pmol x L(-1), P < .05) and a lower suprabasal insulin effect (0.057 +/- 0.03 v 0.023 +/- 0.009 min( 1), P < .05) than control subjects with high tolerance. Offspring with low tolerance had a lower AIRg (2,574 +/- 1,197 v 3,707 +/- 1,656 pmol x L(-1), P < .05) and a lower GEZI (0.101 +/- 0.05 v 0.212 +/- 0.08 x 10(-1) x min(-1), P < .05) than offspring with high tolerance. Offspring with high and low tolerance showed lower phi1 (375 +/- 155 v 272 +/- 181 v 698 +/- 336 (pmol x L(-1))min(mmol x L(-1)), NS) than control subjects with high tolerance. In conclusion, our data suggest that decreases in GEZI and AIRg are the main factors responsible for the worsening of intravenous glucose tolerance in the offspring of NIDDM patients. PMID- 10459562 TI - Effects of low-intensity aerobic training on the high-density lipoprotein cholesterol concentration in healthy elderly subjects. AB - The concentration of high-density lipoprotein cholesterol (HDL-C) is inversely correlated with the risk of coronary heart disease. The effects of low-intensity aerobic training on serum HDL-C and other lipoprotein concentrations were examined in healthy elderly subjects. The subjects were randomly assigned to two groups matched for sex, age, height, and weight. The training group (n = 20, 10 men and 10 women aged 67 +/- 4 years) participated in a supervised physical exercise regimen using a bicycle ergometer at an intensity of 50% estimated maximal oxygen consumption (VO2max) for 60 minutes two to four times per week for 5 months. In contrast, the control group (n = 20, 10 men and 10 women aged 68 +/- 4 years) did not perform any particular physical training. The training protocol resulted in significant increases in the VO2max (P < .05), HDL-C, HDL2-C, and HDL2-C/HDL3-C ratio (P < .01). The change in HDL2-C (r = .57, P < .01) and HDL2 C/HDL3-C (r = .63, P < .01) was positively associated with an increase in the total exercise duration per week. In addition, the total weekly exercise duration also showed a significant positive relationship with HDL-C (r = .75, P < .01), HDL2-C (r = .81, P < .01), and HDL2-C/HDL3-C (r = .71, P < .01) after the training period. The changes in body weight and the VO2max were not significantly correlated with any lipid parameters. Low-intensity aerobic training may improve the profile of HDL-C and its subfractions in healthy elderly subjects. Also, the total exercise duration may be an important factor for improving HDL-C and HDL2-C in elderly subjects. PMID- 10459563 TI - Hyperinsulinemia in a normal population as a predictor of non-insulin-dependent diabetes mellitus, hypertension, and coronary heart disease: the Barilla factory revisited. AB - The study was initiated to evaluate the ability of hyperinsulinemia (as a surrogate measure of insulin resistance) to predict the development in a previously healthy population of three putative outcomes of this abnormality- glucose intolerance, hypertension, and coronary heart disease (CHD). The study involved defining the incidence at which these changes occurred between 1981 and 1993 to 1996 in 647 individuals who were free of any disease when initially studied. The study population consisted of approximately 90% of the subjects evaluated in 1981, divided into quartiles on the basis of the plasma insulin response to a glucose challenge as determined in 1981. The results indicated that the 25% of the population with the highest insulin response in 1981 had significant (P < .001) increases in the incidence of impaired glucose tolerance (IGT) or type 2 diabetes (eightfold), hypertension (twofold), or CHD (threefold). Furthermore, the ability of hyperinsulinemia to predict the three clinical endpoints was independent of differences in age, gender, or body mass index (BMI). Finally, if CHD is considered the clinical endpoint, multiple logistic regression analysis indicates that the values for plasma triglyceride (TG) and mean arterial blood pressure ([MAP] as measured in 1981) also predict the development of CHD. These results indicate that the untoward clinical effects of insulin resistance and/or compensatory hyperinsulinemia, glucose intolerance, hypertension, and CHD clearly can develop in less than 15 years. PMID- 10459564 TI - Insulin has a biphasic effect on the ability of human chorionic gonadotropin to induce ovarian cysts in the rat. AB - Hyperinsulinemia enhances the ability of subovulatory doses of human chorionic gonadotropin (hCG) to induce ovarian follicular cysts in the rat. To determine the relative contribution of these hormones to the development of ovarian cysts, adult female rats were treated with either (1) vehicle alone (controls), (2) a high-fat diet (HFD) to control for the effects of weight gain, (3) 1.5 to 6 IU hCG twice daily plus 6 U insulin (Ins)/d, or (4) 1.5 to 9 U Ins/d plus 3 IU hCG twice daily. On day 23 of the in vivo treatments, all groups that received at least 6 U Ins/d displayed increased body weight compared with control and HFD rats (P < or = .05). No control rats and only one HFD rat displayed ovarian cysts on this day. Plasma estrone (E1) and androstenedione (A4) were elevated in HFD rats with noncystic follicles compared with control rats (P < or = .05). Between 64% and 80% of rats on 6 U Ins/d plus twice-daily injections of 1.5 to 6 IU hCG displayed ovarian cysts on day 23. Plasma estradiol (E2) concentrations for these treatment groups were similar to those of control rats. Of the hormonally treated animals, only those that had ovarian cysts in response to twice-daily injections of 4.5 or 6 IU hCG plus 6 U Ins/d displayed elevated plasma A4 and/or testosterone compared with controls. In contrast, plasma E1 concentrations were elevated on day 23 for animals bearing ovarian cysts in response to increasing doses of hCG plus the fixed dose of 6 U Ins/d. Between 70% and 80% of rats treated twice daily with 3 IU hCG plus a daily dose of 1.5 to 6 U Ins displayed ovarian cysts on day 23. In marked contrast, only 25% of rats treated with this dose of hCG plus 9 U Ins/d developed cystic follicles. Of the plasma steroids tested, only E1 and A4 were elevated in these treatment groups compared with controls. However, these increases in plasma steroid concentrations did not correlate with the dose of insulin. We conclude from these data that, although the mechanisms remain to be elucidated, extreme hyperinsulinemia has the paradoxical ability to attenuate the induction of ovarian cysts by hCG in some animals. PMID- 10459565 TI - Individualized low-dose growth hormone (GH) treatment in GH-deficient adults with childhood-onset disease: metabolic effects during fasting and hypoglycemia. AB - Growth hormone (GH) has insulin-antagonistic effects, and GH secretion is augmented during fasting and hypoglycemia. In the present study, 10 patients aged 21 to 28 years with childhood-onset GH deficiency (GHD) were studied during a 24 hour fast and a hypoglycemic glucose clamp before and after 9 months of GH replacement. During the 24-hour fast, blood glucose, serum insulin, and serum free fatty acid (FFA) levels were measured. In the hypoglycemic clamp, the counterregulatory hormones (plasma catecholamines, serum glucagon, and serum cortisol), serum insulin-like growth factor (IGF) binding protein-1 (IGFBP-1), serum FFA, and glucose uptake were measured. The GH dose was adjusted to the response of serum IGF-I, and the median GH dose was 0.14 IU/kg/wk (range, 0.08 to 0.19). At the end of the study, serum IGF-I levels were normalized in all but one patient, in whom serum IGF-I was above the normal range. Nine months of GH treatment did not cause any significant changes in the blood glucose level, insulin to glucose ratio, or serum FFA level during the 24-hour fast, and none of the patients experienced hypoglycemia either before or after GH treatment. However, GH therapy resulted in increased insulin resistance during hypoglycemia, without changes in the counterregulatory hormonal responses, serum IGFBP-1, or serum FFA. PMID- 10459566 TI - Twenty-four-hour variation in serum leptin in the elderly. AB - To investigate the possibility that the aging process may affect the diurnal variation in serum leptin in humans, serum leptin levels were measured by a sensitive radioimmunoassay method in 12 elderly (aged 72 to 87 years) and 10 middle-aged (35 to 50 years) lean male subjects. Fasting blood samples (4 mL) were drawn at 8:00 AM, and then every 4 hours until 10:00 PM and every 2 hours from 12:00 midnight to 8:00 AM of the next morning. Circadian rhythmicity analysis was performed using the cosinor method. In elderly subjects, serum leptin levels showed a significant diurnal rhythm, which was similar to that observed in controls. Single cosinor analysis showed a significant rhythm in eight of 12 elderly subjects and in all middle-aged subjects but one. Compared with middle-aged subjects, similar mesor mean values (7.8 +/- 1.0 v 8.1 +/- 0.8 ng/mL) but a decreased amplitude (1.4 +/- 0.3 v 2.3 +/- 0.2 ng/mL) and an earlier acrophase (11:56 PM v 2:04 AM) were observed in the elderly. The data demonstrate that the diurnal variation in serum leptin is generally preserved in the elderly. However, the amplitude of leptin diurnal excursion undergoes a reduction with advancing age. It can be speculated that the blunted diurnal variation in serum leptin observed in the elderly may result in an alteration of the afferent signal in the adipose tissue-central nervous system homeostatic loop. PMID- 10459567 TI - Elevated glucose potentiates contraction of isolated rat resistance arteries and augments protein kinase C-induced intracellular calcium release. AB - The effect of elevated glucose on arterial contractions and intracellular calcium ([Ca++]i) release induced by protein kinase C (PKC) activation and potassium depolarization (KCl) was investigated. Mesenteric resistance arteries (phi < 200 microm) isolated from male Wistar rats were studied using an arteriograph system that allowed control of transmural pressure (TMP) and measurement of lumen diameter. Arteries were incubated in either 11 or 44 mmol/L glucose and the concentration-response to Indolactam V (ILV; a specific PKC activator; LC Laboratories, Woburn, MA) and KCl was determined, as well as the sensitivity to Ca++ in the presence of either agonist. An additional group of arteries were incubated in 5.5 mmol/L glucose and the concentration-response to ILV was compared versus 11 and 44 mmol/L glucose. Arteries in 44 mmol/L glucose were more sensitive to both ILV and KCl, contracting to 10.0 micromol/L ILV, 53.9 +/- 10.1% in 11 mmol/L versus 85.1 +/- 2.0% in 44 mmol/L glucose (P < .01); arteries in 5.5 mmol/L glucose responded the least to ILV, contracting only 36.0 +/- 4% to 10.0 micromol/L ILV (P < .01 v 11 and 44 mmol/L glucose). The KCl EC50 (ie, the value at which the agonist produced 50% maximal contraction) for 11 versus 44 mmol/L glucose was 41.3 +/- 4.8 versus 31.1 +/- 1.2 mmol/L (P < .05). There was no change in Ca++ sensitivity in elevated glucose for either agonist; however, Ca++ sensitivity was augmented threefold for ILV versus KCl, demonstrating an agonist dependent modulation of Ca++ sensitivity. The Ca++ EC50 for ILV and KCl in 11 versus 44 mmol/L was 0.18 +/- 0.05 versus 0.21 +/- 0.05 and 0.59 +/- 0.09 versus 0.60 +/- 0.10 micromol/L (P < .01 v ILV). The effect of elevated glucose on [Ca++]i release from the sarcoplasmic reticulum (SR) was investigated in arteries incubated in zero Ca++ buffer containing either 11 or 44 mmol/L glucose by measuring the contraction produced by 50 mmol/L caffeine, 3.0 micromol/L ILV, or 60 mmol/L KCl. Contraction to caffeine in 11 versus 44 mmol/L glucose was comparable, constricting 42.0 +/- 6.0% in 11 mmol/L and 36.0 +/- 4.4% in 44 mmol/L glucose (P > .05), and contraction to KCl was almost undetectable in both glucose concentrations. However, contraction to ILV increased from 5.6 +/- 0.9% in 11 mmol/L to 18.7% +/- 2.2% in 44 mmol/L glucose (P < .01), indicating that although the amount of Ca++ in the SR (caffeine-sensitive stores) was not increased in elevated glucose, PKC-induced release of [Ca++]i was enhanced, a consequence that may explain the noted glucose-induced increase in contraction to PKC activation. PMID- 10459568 TI - Effect of amino acids on the plasma concentration and urinary excretion of uric acid and uridine. AB - To determine the effect of amino acids on the plasma level and urinary excretion of uric acid and uridine, 200 mL 12% amino acid solution, and 2 weeks later, 100 mL physiological saline solution containing glucagon (1.2 microg/kg weight), was infused into five healthy men. Both increased the urinary excretion of uric acid and the concentration of glucagon, insulin, and glucose in plasma and pyruvic acid in blood, whereas they decreased the concentration of uridine and inorganic phosphate in plasma. However, neither the amino acid infusion nor glucagon infusion affected the concentration of purine bases (hypoxanthine, xanthine, and uric acid), cyclic adenosine monophosphate (cAMP) in plasma, or lactic acid in blood or the urinary excretion of oxypurines (hypoxanthine and xanthine), uridine, or sodium. These results suggest that glucagon may have an important role in the amino acid-induced increase in urinary excretion of uric acid and decrease in plasma uridine. PMID- 10459569 TI - Longitudinal changes in serum 25-hydroxyvitamin D in older people. AB - Cross-sectional studies have suggested that serum 25-hydroxyvitamin D (25OHD) levels decline with aging. We have examined this putative decline in a longitudinal study using participants in the New Mexico Aging Process Study. 25OHD levels were measured in participants in whom serum samples were available between 1980 to 1982 and 1989 to 1994 (37 men and 99 women). The available data for these visits included age, gender, and the date the sample was obtained. Questionnaires assessing physical activity and vitamin D intake were administered at the visits. A seasonal variation (r = .25, P < .05) in 25OHD was demonstrated in the whole group of subjects. In 25 subjects who were not receiving vitamin D supplementation at either time and had samples obtained in the same season, both serum 25OHD (P < .05) and physical activity (P < .05) decreased over a mean period of 11.4 years. In 23 subjects who had samples obtained in the same season but used vitamin D supplements at both times, there was no change in serum 25OHD. Mean summer 25OHD levels did not change with the duration of study. On the other hand, the mean serum 25OHD declined with the duration of study when measured from winter to winter or spring to spring. Multiple regression analysis demonstrated that the month, activity level, vitamin D supplementation, and gender (P < .001) were independent determinants of serum 25OHD levels. This study confirms that aging is associated with a reduction in serum 25OHD, and suggests that this decrease is a reflection of reduced sun exposure rather than aging per se. The reduction in serum 25OHD was the result of decreasing winter and spring 25OHD serum concentrations. It is clear that vitamin D supplementation can prevent the age-related decline in 25OHD levels. PMID- 10459570 TI - Bromocriptine/SKF38393 treatment ameliorates dyslipidemia in ob/ob mice. AB - Our previous studies have shown that the dopaminergic D1 receptor agonist SKF38393 (SKF) plus the D2 receptor agonist bromocriptine (BC) act synergistically to reduce obesity in obese C57BL/6J (ob/ob) mice. The present study investigated the effects of this combination on dyslipidemia in ob/ob mice. Female ob/ob mice were treated daily with vehicle or SKF (20 mg/kg body weight [BW]) plus BC (16 mg/kg BW [BC/SKF]) for 14 days. The animals were then used for the characterization of plasma lipoprotein profiles, hepatic triacylglycerol synthesis and secretion, adipocyte lipolysis, adipose and muscle lipoprotein lipase (LPL) activity, and muscle triglyceride (TG) content. The treatment significantly reduced plasma glucose 54%, TG 41%, cholesterol 21%, phospholipid 20%, and free fatty acid (FFA) 36% (P < .01). Hepatic triacylglycerol synthesis was 55% lower in treated mice versus control mice (P < .01). The cell size of isolated adipocytes was significantly reduced (41%) by treatment. LPL activity was increased in soleus skeletal muscle (25%, P < .05) but was sharply reduced in adipose tissue (91%, P < .01) in treated versus control mice. The TG content of hindlimb muscle was about 49% lower in treated versus control mice (P < .05). The basal and isoproterenol-stimulated lipolytic rate was decreased (approximately 53%) in adipocytes from treated animals compared with the control (P < .01). In conclusion, BC/SKF normalized the hypertriglyceridemia likely via its simultaneous antilipogenic action in liver tissue and antilipolytic action in adipose tissue. Decreased plasma flux of FFA partially contributed to the reduced hepatic lipogenesis, plasma very-low-density lipoprotein (VLDL)-TG, and TG in skeletal muscle. The above-described effects of BC/SKF treatment are largely independent of its effect to normalize hyperphagia in ob/ob mice. PMID- 10459571 TI - Insulin resistance in adipocytes from spontaneously hypertensive rats: effect of long-term treatment with enalapril and losartan. AB - Insulin responsiveness was studied in isolated adipocytes from the normotensive Wistar Kyoto (WKY) rat and the spontaneously hypertensive rat (SHR). The effect of insulin (0.1 to 5 nmol/L) on glucose uptake (glucose transport and lipogenesis) was measured, and the maximal effect of insulin (Emax) and the dose of insulin required to elicit 50% of the maximal response (EC50) were calculated. A diminished Emax on lipogenesis without changes in the EC50 was detected in SHRs. The Emax was 0.49 +/- 0.09 (SHR) and 1.16 +/- 0.14 (WKY) micromol/10(5) cells (P < .05), and the EC50 was 0.13 +/- 0.03 and 0.11 +/- 0.02 nmol/L for WKY and SHR, respectively. Similar results were obtained when measuring insulin stimulated glucose transport. A 30-day long-term treatment with enalapril (20 mg/kg/d) normalized insulin responsiveness in adipocytes from SHRs. The effect of enalapril was suppressed when SHRs were pretreated with enalapril and 150 microg/kg/d of the bradykinin (BK) B2-receptor blocker Hoe 140. Pretreatment with losartan (40 mg/kg/d) did not improve insulin action in the SHR. Since these results were obtained with isolated cells in which glucose availability was not a function of blood flow, and the effect of insulin in the SHR was improved by pretreatment with an angiotensin-converting enzyme (ACE) inhibitor but not with the AT1-receptor blocker, it appears that the insulin resistance linked to the hypertension is not related to changes in blood flow. PMID- 10459572 TI - A common mutation in the methylenetetrahydrofolate reductase gene is a determinant of hyperhomocysteinemia in epileptic patients receiving anticonvulsants. AB - Hyperhomocysteinemia is a condition caused by both genetic and nongenetic factors. To determine whether a common methylenetetrahydrofolate reductase (MTHFR) variant is related to elevated homocysteine concentrations in epileptic patients receiving anticonvulsants, we investigated the plasma total homocysteine (tHcy) level, folate level, and MTHFR 677 C --> T mutation using a polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis with HinfI digestion in 103 patients with epilepsy and 103 normal controls. The prevalence of hyperhomocysteinemia (> or = 11.4 micromol/L, 90th percentile of control group) was higher in patients than in controls (25% v 10.0%, P = .007). The homozygosity for the 677 C --> T mutation of MTHFR was associated with elevated tHcy and low folate levels. The magnitude of hyperhomocysteinemia in MTHFR TT homozygotes was more pronounced in epileptic patients than in controls (18.2 +/- 1.6 v 9.1 +/- 1.2 micromol/L, P = .04). In epileptic patients, hyperhomocysteinemia was more frequent in MTHFR TT genotypes versus CT or CC genotypes (58% v 17% and 16%, P < .001). Multiple logistic regression analysis showed that MTHFR TT genotype was an independent predictor of hyperhomocysteinemia in epileptic patients receiving anticonvulsants (phenytoin and carbamazepine but not valproic acid), suggesting that gene-drug interactions induce hyperhomocysteinemia. These findings indicate that epileptic patients receiving anticonvulsants may have a higher folate requirement to maintain a normal tHcy level, especially homozygotes for MTHFR 677 C --> T mutation. PMID- 10459573 TI - Glucose-6-phosphatase mRNA levels in kidney isolated tubule suspensions are increased by dexamethasone and decreased by insulin. AB - The strong induction of renal glucose-6-phosphatase (G6Pase) during starvation has been suggested to be responsible for the increased role of the kidney in glucose production during long-term fasting. To investigate whether this induction may be caused by a direct hormonal effect on the renal proximal tubular cell, we incubated rat renal tubule suspensions in the presence of glucocorticoids or insulin for 6 hours; normoxia was required, since hypoxic conditions were associated with markedly decreased G6Pase mRNA levels despite maintenance of adenosine triphosphate (ATP) levels. The G6Pase mRNA level was increased twofold to threefold by 10(-8) to 10(-5) mol/L dexamethasone (DXM), whereas the most effective concentration of insulin, 10(-9) mol/L, induced only a 40% decrease. These results suggest that the increased role of the kidney in glucose production during long-term starvation could be linked to a direct effect of glucocorticoids on renal G6Pase. PMID- 10459574 TI - Very-low-density lipoprotein apolipoprotein B100 kinetics in adult hypopituitarism. AB - Hypopituitarism is associated with hyperlipidemia, the mechanisms of which are not fully known. One possible mechanism is an increased hepatic secretion of very low-density lipoprotein (VLDL) apolipoprotein B100 (apo B100). To investigate this, 13 hypopituitary patients (seven women and six men; age, 46 +/- 3 years [mean +/- SEM]; body mass index [BMI], 29 +/- 2 kg/m2) and 13 matched controls (seven women and six men; age, 43 +/- 3 years; BMI, 28 +/- 2 kg/m2) were investigated in a stable-isotope study. [1-(13)C]leucine (1 mg/kg body weight) was administered, followed by a continuous 6-hour infusion of [1-(13)C]leucine (at a rate of 1 mg/kg/h). Patients had a similar fractional secretion rate (FSR) of VLDL apo B100 versus controls (0.37 +/- 0.05 v 0.38 +/- 0.06 pools/h, respectively), but they had a significantly larger pool size (3.4 +/- 0.3 v 1.9 +/- 0.3 mg/kg) and higher absolute secretion rate ([ASR] 27.8 +/- 2.9 v 16.0 +/- 2.5 mg/kg/d). The increase in hepatic VLDL production may explain the lipid abnormalities found in hypopituitarism. Fasting circulating nonesterified fatty acids (NEFAs) were decreased in the patients (284 +/- 26 v 664 +/- 92 micromol/L, P < .001) despite the increase in VLDL secretion. An inverse relationship was observed between the NEFA level and VLDL apo B100 FSR in the patients (r(s) = .85, P < .005). PMID- 10459575 TI - Overproduction of insulin in the chromium-deficient rat. AB - The hypothesis that the insulin secretory hyperresponsiveness observed in rats with diet-induced insulin resistance may be a basic characteristic of dietary chromium (Cr) deficiency was evaluated. Two groups of weanling rats were fed ad libitum a purified diet containing 64% sucrose, 20% casein, 5% corn oil, and the recommended levels of vitamins and minerals without added Cr. Cr-deficient (-Cr) rats were provided with distilled drinking water only, while Cr-supplemented (+Cr) rats received water containing 5 ppm Cr as CrCl3. A third group of rats fed a commercial chow diet served as sucrose controls. Effects of Cr deficiency were assessed by comparing fasting levels of glucose, insulin, and plasma lipids in blood samples collected biweekly from the -Cr and +Cr groups over a 3-month period. Both groups of rats fed the low-Cr sucrose diet developed a transient hyperinsulinemia and hyperlipidemia relative to the chow-fed control rats. There were significant effects of Cr supplementation on plasma triglycerides during the initial 2 weeks of dietary adaptation. Effects of the low-Cr diet were evaluated after the 12-week period by comparing the insulin response area and glucose clearance during a 40-minute intravenous glucose tolerance test (IVGTT). The rates of glucose clearance (KG) in -Cr and +Cr rats were similar (4.2 +/- 1.0 and 4.3 +/- 0.8%/min, respectively) and were comparable to the K(G) in chow-fed rats (4.6 +/- 0.8). In contrast, insulin secretory responses in -Cr rats were exaggerated (area, 14,083 +/- 3,399 microU/mL x min), being twofold greater (P < .05) relative to the +Cr group (6,183 +/- 864). The insulin secretory response area in chow-fed rats (7,081 +/- 408 microU/mL x min) was similar to the value in the +Cr group. These observations provide support for the hypothesis that Cr deficiency can lead to elevated insulin secretory responses to glucose. PMID- 10459576 TI - Zinc supplementation improves glucose disposal in patients with cirrhosis. PMID- 10459577 TI - Aerosolised liposomal hamycin for treatment of systemic Candida infections in mice. AB - Mice lethally infected with Candida albicans were exposed to small-particle aerosols containing hamycin liposomes. The drug, when administered twice daily for 2 h (0.68 mg kg(-1) body weight per day) on days 1, 2 and 3 post inoculation, significantly reduced the numbers of Candida organisms in the kidneys. Aerosol treatment increased the survival time of mice given two 2-h treatments once a week for 4 weeks. A twice weekly 2-h small particle aerosol administration of hamycin for 1, 2, or 3 weeks significantly increased both the mean time of survival and percent survival. PMID- 10459578 TI - Studies on the functional site on staphylococcal enterotoxin A responsible for production of murine gamma interferon. AB - To identify the functional region(s) associated with induction of gamma interferon on the staphylococcal enterotoxin A molecule, native staphylococcal enterotoxin A molecules and 12 various synthetic peptides corresponding to different regions of entire staphylococcal enterotoxin A were compared to induce gamma interferon production in murine spleen cells. The native staphylococcal enterotoxin A molecule induced gamma interferon production, whereas all of the 12 synthetic peptides did not. Pre-treatment of the murine spleen cells with synthetic peptide A-9 (corresponding to amino acid residues 161-180) significantly inhibited the staphylococcal enterotoxin A-induced gamma interferon production, whereas those with other synthetic peptides did not. When native staphylococcal enterotoxin A was pre-treated with either anti-staphylococcal enterotoxin A serum or anti-peptide sera, anti-staphylococcal enterotoxin A serum and antisera to peptides A-1 (1-20), A-7 (121-140), A-8 (141-160), A-9 (161-180) and A-10 (181-200) inhibited the staphylococcal enterotoxin A-induced gamma interferon production. From these findings, the amino acid residues 161-180 on the staphylococcal enterotoxin A molecule may be an essential region for murine gamma interferon production. Furthermore, the neutralizing epitopes may be also located on regions of amino acid residues 1-20, 121-140, 141-160 and 181-200 on the staphylococcal enterotoxin A molecule. PMID- 10459579 TI - Cloning and characterization of a gene encoding an antigenic membrane protein from Actinobacillus pleuropneumoniae with homology to ABC transporters. AB - Actinobacillus pleuropneumoniae is a pathogenic bacterium responsible for a highly contagious and often fatal form of bronchopneumonia in swine. Survival from a natural infection generally results in immunity from further infection by all 12 common serotypes, suggesting the presence of common protective antigens. We have identified one of the antigenic membrane proteins from A. pleuropneumoniae serotype 5, and cloned the gene which encodes it. This gene is found in all 12 serotypes, and encodes a protein with a predicted molecular mass of 30 kDa. Sequence analysis revealed that this antigen has a typical signal sequence characteristic of lipoproteins, and is likely to be secreted and inserted into the periplasmic side of the inner membrane. The gene shows high homology to the surface antigen CjaA of Campylobacter jejuni and to solute binding proteins of the ABC transporter family. The probable role of this protein in substrate binding and transport was supported by the presence of an upstream gene with significant homology to ATP binding proteins of the same family. In Escherichia coli, the cloned gene produced a protein which reacted strongly with convalescent sera from swine infected with A. pleuropneumoniae serotype 5, and weakly with sera from swine infected with serotype 1A or from swine vaccinated with a killed bacterin of serotype 1A or 5. It thus appears that this antigen displays some crossreactivity between serotypes, and may be less exposed in bacterins than in live cells. This protein, designated ApaA, may have an important role in nutrient acquisition and in the pathogenesis of infections caused by A. pleuropneumoniae. PMID- 10459580 TI - Sequences from the aspergillopepsin PEP gene of Aspergillus fumigatus: evidence on their use in selective PCR identification of Aspergillus species in infected clinical samples. AB - In immunodeficient patients, Aspergillus species emerge as circumstantial pathogens. Aspergillus fumigatus is a distant first among the pathogenic aspergilli, which cause deep-seated mycoses. Sequences of the pep gene of A. fumigatus as potential PCR primers, which have not been tested before, were used to identify this species and if possible, differentiate it from other, co identified, clinically important species of the genus. We present results of the three most promising primer pairs, pep-1/pep-22, pep-15/pep-22 and pep-21/pep32. The second pair was of better specificity when tested with DNA extracted from pure cultures of a multitude of aspergilli, whereas the first co-amplified four clinically significant Aspergillus species. The compatibility of the PCR method with the CTAB DNA extraction protocol varied according to the biological fluid tested and the primer pair used. The first two pairs showed moderate adaptability to the different commercial DNA extraction kits, which were tested in whole blood, spiked with Aspergillus fumigatus hyphae and conidia - as were all the biological fluids used. Restriction of the amplification products with MspI produced distinct patterns for different Aspergillus spp. This approach, as a potential diagnostic tool, seems reliable and sensitive due to its flexibility, speed, low cost, ease of application and selectable breadth of detection. PMID- 10459581 TI - F165(1) fimbriae of the P fimbrial family inhibit the oxidative response of porcine neutrophils. AB - The F165(1) fimbrial system has been associated with the resistance of Escherichia coli O115:K"V165" to phagocytic killing by porcine polymorphonuclear leukocytes (PMNLs). One mechanism of this resistance seemed to be inhibition of the oxidative response as observed following induction of PMNLs by phorbol myristate acetate (PMA) and treatment with bacteria possessing the F165(1) fimbriae. In order to confirm whether or not the F165(1) fimbriae are involved in this inhibition, we evaluated the effect of F165(1)-positive strains (a pathogenic wild-type strain 5131, and a recombinant strain HB101(pCJ7)) or an F165(1)-negative strain HB101 (used as negative control) on the oxidative response of porcine neutrophils (pNs) stimulated with PMA. Incubation of pNs with pathogenic E. coli strain 5131 resulted in significant inhibition of the oxidative response as compared to that observed for pNs incubated without bacteria, as assessed by hydrogen peroxide (H2O2) and superoxide anion (O2-) release from the phagocytes, and by the chemiluminescence assay. Similarly, incubation of pNs with the F165(1)-producing cloned strain HB101(pCJ7) resulted in significant inhibition of the pN oxidative response as compared to that observed for pNs incubated without bacteria or with strain HB101. In contrast, addition of purified F165(1) fimbriae to the pNs had no effect on the oxidative response. PMID- 10459582 TI - Electrotransformation of the human pathogenic fungus Scedosporium prolificans mediated by repetitive rDNA sequences. AB - The regions encoding the 5.8S rRNA and the flanking internal transcribed spacers (ITSI and ITSII) from two isolates of the human pathogenic fungus Scedosporium prolificans and one isolate of the taxonomically related species Pseudallescheria boydii (S. apiospermum) were sequenced. The sequences of the two S. prolificans isolates were identical. However, there were minor differences between both species. Phylogenetic analysis of known fungal sequences confirmed a close relationship between S. prolificans and P. boydii. An attempt was made to transform S. prolificans by electroporation using a plasmid vector, pMLF2, bearing the Escherichia coli hygromycin B phosphotransferase gene (hph) under the control of Aspergillus nidulans promoter and terminator sequences. To increase transformation efficiency, the sequenced ribosomal cluster of S. prolificans was used to construct a new vector for homologous recombination. PMID- 10459583 TI - Immune response to rotavirus VP4 expressed in an attenuated strain of Shigella flexneri. AB - An attenuated strain of Shigella flexneri was utilised to express viral protein (VP) 4 of rotavirus and the immunogenicity of the recombinant constructs was studied in BALB/c mice. VP4 was expressed as a fusion with maltose binding protein (MBP) in both the cytoplasm and periplasm, with a much higher level of expression occurring in the former. While all constructs induced a Shigella specific response in mice, only the construct expressing MBP-VP4 in the cytoplasm of Shigella stimulated an immune response specific to rotavirus. This study demonstrates that Shigella can be used to deliver rotavirus antigens and induces an immune response directed towards both rotavirus and Shigella. PMID- 10459584 TI - A surface-displayed cholera toxin B peptide improves antibody responses using food-grade staphylococci for mucosal subunit vaccine delivery. AB - The possibility of improving the antibody responses to a model streptococcal antigen, administered by intranasal immunization as surface-displayed on the food grade bacterium Staphylococcus carnosus, by co-exposure of a peptide (CTBp) comprising amino acids 50-75 of the cholera toxin B subunit, was investigated. It was found that the introduction of the CTBp into the chimeric surface proteins, containing a serum albumin binding protein (ABP) from streptococcal protein G as model antigen, significantly increased serum IgG responses upon intranasal immunization. Similarly, elicited local IgA responses were also found to be improved. Furthermore, it was demonstrated that live delivery of the staphylococci was required to obtain this effect, since UV-irradiated or heat killed bacteria exposing the same chimeric surface proteins did not show increased anti-ABP IgG responses. PMID- 10459585 TI - Toxoplasma gondii infection: analysis of serological response by 2-DE immunoblotting. AB - Toxoplasma gondii is known to cause a variety of diseases ranging from asymptomatic infections to serious conditions in immunocompromised hosts such as AIDS-patients or transplant recipients. In addition they may cause abortion or fetal abnormalities during pregnancy. Despite the clinical importance, diagnosis, treatment and prevention still remain unsatisfactory. Analysis of the parasitic cell determinants, recognized by specific humoral and cellular immune responses, may have important implications for diagnosis, therapy and vaccination strategies. Two-dimensional electrophoresis (2-DE) was used to resolve and compare protein patterns from Toxoplasma gondii strains RH and BK (mouse virulent strains). Comparison of silver-stained gels showed that 35.2% to 60.3% of the spots had the same position. In a second series of experiments, the reactivity of the spots with human sera was tested. Proteins were transferred to PVDF membranes and were detected with sera from different patient groups. Depending upon the immunoglobulin class (IgG, IgM, IgA or IgE) different epitope patterns were observed. Some of the spots seemed to be recognized in different stages of infection. Sera of two patients with similar serology and comparable stage of infection were compared in order to demonstrate an individual immune response. PMID- 10459586 TI - The role of Pseudomonas aeruginosa elastase as a potent inflammatory factor in a rat air pouch inflammation model. AB - Pseudomonas aeruginosa, an opportunistic pathogen, can cause life threatening infections in patients compromised by underlying respiratory disease like bronchiectasis, cystic fibrosis and diffuse panbronchiolitis. Most strains of P. aeruginosa produce some kind of protease with broad substrate specificities during the infectious state in the host. P. aeruginosa elastase, one of the strongest exotoxins, has a tissue-damaging proteolytic activity and is capable of degrading such plasma proteins as immunoglobulins, complement factor and cytokines. The present study focused on the effect of P. aeruginosa elastase and was designed to evaluate the neutrophil accumulation at the inflammation site mediated by P. aeruginosa elastase in the inflammatory response in the host. An air pouch model in rats, considered as a useful model of inflammation, was used to analyze the number of leukocytes, the volume of exudate and the concentration of interleukin-8 after the injection of P. aeruginosa elastase into the pouch cavity. The number of neutrophils and the volume of exudate in the pouch cavity increased significantly at 4 h, peaked at 8 h in a dose-dependent manner and then decreased at 24 h. The concentration of interleukin-8 in pouch fluid peaked 4 h earlier than the peak of the neutrophil number. The enzymatic activity of P. aeruginosa elastase seemed to reinforce the inflammation process. The influence of lipopolysaccharide contamination was negligible. Although these observations were made in the subcutaneous cavity, they indicate that P. aeruginosa elastase plays a role as an immunoprovocative factor in the inflammatory response in cases of infection with P. aeruginosa. PMID- 10459587 TI - Life supporting first aid training of the public--review and recommendations. AB - Since the introduction around 1960 of external cardiopulmonary resuscitation (CPR) basic life support (BLS) without equipment, i.e. steps A (airway control)-B (mouth-to-mouth breathing)-C (chest (cardiac) compressions), training courses by instructors have been provided, first to medical personnel and later to some but not all lay persons. At present, fewer than 30% of out-of-hospital resuscitation attempts are initiated by lay bystanders. The numbers of lives saved have remained suboptimal, in part because of a weak or absent first link in the life support chain. This review concerns education research aimed at helping more lay persons to acquire high life supporting first aid (LSFA) skill levels and to use these skills. In the 1960s, Safar and Laerdal studied and promoted self-training in LSFA, which includes: call for the ambulance (without abandoning the patient) (now also call for an automatic external defibrillator); CPR-BLS steps A-B-C; external hemorrhage control; and positioning for shock and unconsciousness (coma). LSFA steps are psychomotor skills. Organizations like the American Red Cross and the American Heart Association have produced instructor-courses of many more first aid skills, or for cardiac arrest only-not of LSFA skills needed by all suddenly comatose victims. Self-training methods might help all people acquire LSFA skills. Implementation is still lacking. Variable proportions of lay trainees evaluated, ranging from school children to elderly persons, were found capable of performing LSFA skills on manikins. Audio-tape or video-tape coached self-practice on manikins was more effective than instructor-courses. Mere viewing of demonstrations (e.g. televised films) without practice has enabled more persons to perform some skills effectively compared to untrained control groups. The quality of LSFA performance in the field and its impact on outcome of patients remain to be evaluated. Psychological factors have been associated with skill acquisition and retention, and motivational factors with application. Manikin practice proved necessary for best skill acquisition of steps B and C. Simplicity and repetition proved important. Repetitive television spots and brief internet movies for motivating and demonstrating would reach all people. LSFA should be part of basic health education. LSFA self-learning laboratories should be set up and maintained in schools and drivers' license stations. The trauma focused steps of LSFA are important for 'buddy help' in military combat casualty care, and natural mass disasters. PMID- 10459588 TI - Basic life support training for health care students. AB - This paper describes a novel method for delivering basic life support training to undergraduate healthcare students. A comprehensive 8 h programme is organised and delivered by undergraduate students to their peers. These students have undergone training as basic life support instructors validated by the Royal Life Saving Society UK. The course is delivered to multiprofessional groups of medical, dental, physiotherapy, biomaterial and nursing undergraduates. It has been well received by students and academic staff and provides a solution to reduce the workload of over burdened clinical staff while at the same time enhancing quality. It forms part of an overall strategy for improving resuscitation training for undergraduates from all disciplines. PMID- 10459589 TI - Long-term survival after out-of-hospital cardiac arrest: an 8-year follow-up. AB - Between 1988 and 1994, 441 patients were successfully resuscitated outside hospital in the city of Rotterdam, of whom 276 (63%) were discharged from hospital alive. Long-term survival was studied amongst those who were discharged alive. The duration of follow-up averaged 6.71 years. A survival rate of 88% after 1 year, 81% after 3 years, 77% after 5 years and 73% after 7 years was found. After multivariate analysis, age, diagnosis and gender were found to be independent and significant predictors of survival. No significant difference in survival was found in patients who had been resuscitated by emergency personnel, physicians and bystanders. Patients who were still alive were sent a EuroQol questionnaire. No differences in outcomes between the four groups were found. Since long-term prognosis after out-of-hospital resuscitation is satisfactory, learning programmes for resuscitation should be continued. PMID- 10459590 TI - Increased frequency of thorax injuries with ACD-CPR. AB - A prospective, randomised out-of-hospital study in a two-tiered system with active compression-decompression (ACD) cardiopulmonary resuscitation (CPR) versus standard (STD) CPR in patients following non-traumatic cardiac arrest was planned to test the hypothesis that ACD-CPR by the first tier may increase the occurrence of ventricular fibrillation as compared with STD-CPR. Furthermore, in a later phase of the study, sternal and rib fractures induced by both CPR methods were determined by extensive autopsy. After enrolling 90 patients the study was terminated because of a high frequency of chest injuries found at autopsy. Forty two patients received STD-CPR from the first tier and ACD-CPR from the second tier. Thirty-three patients received ACD-CPR only by the first and the second tier, while 15 patients received STD-CPR only from the first and second tiers. In order to obtain a sufficiently large control group for autopsy findings after STD CPR, STD-CPR was performed in an additional 33 patients within a second period of 4 months. There was no improvement in the number of patients found in ventricular fibrillation after ACD-CPR as compared to STD-CPR performed by the first tier. In patients undergoing autopsy (n = 35) there were significantly more sternal fractures with ACD-CPR versus STD-CPR (14/15 vs. 6/20; P <0.005) and rib fractures (13/15 vs. 11/20; P < 0.05) In conclusion, ACD-CPR appears to cause more CPR-related injuries than does standard CPR, but as a result of a number of limitations on this study, this fact cannot be proven beyond doubt. PMID- 10459591 TI - Aminophylline in undifferentiated out-of-hospital asystolic cardiac arrest. AB - PRIMARY OBJECTIVE: To determine if the introduction of intravenous aminophylline, a nonspecific adenosine receptor antagonist, into the resuscitation algorithm of asystole will increase return of spontaneous circulation when used in undifferentiated prehospital cardiac arrest. METHODS: An urban, prehospital, prospective, randomized, double-blind, placebo-controlled trial of nonpregnant normothermic adults suffering nontraumatic out-of-hospital asystolic cardiac arrest. Subjects were treated in accordance with published advanced cardiac life support guidelines and standard pharmacotherapy. They were randomly assigned to receive either placebo or aminophylline along with the initial boluses of atropine and epinephrine. Cardiac rhythms and carotid pulses were monitored throughout the resuscitation. RESULTS: Eighty-two patients were entered into the trial. Forty-five patients were assigned to the placebo group and 37 received aminophylline. Nine of 45 controls (20%; 95% CI 10-35%) achieved return of spontaneous circulation compared to ten of 37 (27%; 95% CI 14-44%) in the aminophylline group. CONCLUSIONS: We were not able to show a statistically significant improvement in return of spontaneous circulation when aminophylline was given during the early resuscitation phase of undifferentiated asystolic cardiac arrest in the prehospital setting with this sample size. PMID- 10459592 TI - The efficacy of atropine in the treatment of hemodynamically unstable bradycardia and atrioventricular block: prehospital and emergency department considerations. AB - OBJECTIVE: To determine the efficacy of atropine therapy in patients with hemodynamically compromising bradycardia or atrioventricular block (AVB) in the prehospital and emergency department settings. METHODS: DESIGN: Retrospective review of prehospital, emergency department, and hospital records. PARTICIPANTS: Prehospital patients with hemodynamically compromising bradycardia or AVB with evidence of spontaneous circulation who received atropine as delivered by emergency medical services personnel (advanced life support level). SETTING: Urban/suburban fire department-based emergency medical service system with on line medical control serving a population of approximately 1.6 million persons. DEFINITIONS: Hemodynamic instability was defined as the presence of any of the following: ischemic chest pain, dyspnea, syncope, altered mental status, and systolic blood pressure less than 90 mmHg. Bradycardia was defined as sinus bradycardia, junctional bradycardia, or idioventricular bradycardia (grouped as bradycardia) while AVB included first-, second- (types I and II), or third-degree (grouped as AVB). The response that occurred within one minute following each dose of atropine was defined as none, partial, complete, or adverse. MAIN RESULTS: Of 172 patients meeting entry criterion complete data was available for 131 (76.1%) and constitutes the study population. The mean age was 71 years. Fifty-one percent were female. Forty-five patients had AVB and 86 bradycardia. Patients with AVB were more likely to have a presenting systolic blood pressure less than 90 mmHg than those with bradycardia. In the 131 patients, responses to atropine were as follows: 26 (19.8%) = partial, 36 (27.5%) = complete, 65 (49.6%) = none, and 4 (2.3%) = adverse. Patients presenting with bradycardia (compared to AVB) more commonly: (1) received a single dose of atropine; (2) a lower total dose of atropine in the prehospital interval; (3) were more likely to arrive in the ED with a normal sinus rhythm; and (4) were less likely to receive additional atropine or isoproterenol in the ED. Those patients who achieved normal sinus rhythm over the total course of care were likely to have achieved that rhythm during the prehospital interval. There was no difference between groups in the likelihood of leaving the ED with a normal sinus rhythm achieved during the ED interval. Acute myocardial infarction was more common in patients presenting with AVB (55.5%) than with bradycardia (23.2%, P = 0.001). CONCLUSIONS: Approximately one-half of patients who received atropine in the prehospital setting for compromising rhythms had either a partial or complete response to therapy. Adverse responses were uncommon. Those patients who presented with hemodynamically unstable bradycardia to EMS personnel responded more commonly to a single dose and a lower total dose of atropine compared to similar patients with AVB. Those patients who achieve normal sinus rhythm by ED discharge were likely to have achieved it during the prehospital interval. PMID- 10459593 TI - Atropine pharmacokinetics and pharmacodynamics following endotracheal versus endobronchial administration in dogs. AB - Emergency endotracheal and endobronchial drug administration provide an effective alternative for intravenous drug delivery during cardiopulmonary resuscitation. The purpose of the present study was to determine the immediate pharmacokinetic and pharmacodynamic properties of atropine following administration by either of these routes. Atropine (0.02 mg/kg) was given to seven anaesthetized mongrel dogs. Each dog was studied twice: once when atropine was injected into the endotracheal tube, and on another day when atropine was given via a flexible catheter wedged into a peripheral bronchus. Plasma atropine concentrations and blood gases were measured during 60 min following drug administration. Both routes of atropine administration differed significantly in three measures: the maximal atropine concentration (Cmax) was significantly higher with the endobronchial administration 40.0 +/- 7.8 ng/ml compared to 23.9 +/- 5 ng/ml endotracheally (P = 0.008); atropine's elimination (t1/2beta) half-life was significantly longer with the endobronchial route (39.3 +/- 5.2 min vs. 28.0 +/- 7.9 min; P = 0.05); Endobronchial administration resulted in an increase of 16% in heart rate, beginning immediately after drug delivery and peaking after 5 min. Other pharmacokinetic parameters were not significantly different. We conclude that endobronchial administration of atropine has a clear advantage over the endotracheal route. PMID- 10459594 TI - Airway management during cardiopulmonary resuscitation--a comparative study of bag-valve-mask, laryngeal mask airway and combitube in a bench model. AB - Gastric inflation and subsequent regurgitation are a potential risk of ventilation during cardiopulmonary resuscitation (CPR). In respect of recent investigations, principal respiratory components such as respiratory system compliance, resistance and lower esophageal sphincter pressure were adapted according to CPR situations. The purpose of our study was to assess lung ventilation and gastric inflation when performing ventilation with bag-valve mask, laryngeal mask airway, and combitube in a bench model simulating an unintubated cardiac arrest patient. Twenty-one student nurses, without any experience in basic life support measures, ventilated the bench model with all three devices. Mean ( +/- S.D.) gastric inflation with the laryngeal mask airway (seven cases) was significantly lower than with the bag-valve-mask (0.6 +/- 0.8 vs 3.0 +/- 2.11 min(-1), P < 0.01). There was no gastric inflation when ventilation was performed with the combitube. Only seven of 21 volunteers exceeded 1-min lung volumes of > 5 1 when using the bag-valve-mask, whereas mean (+/-S.D.) 1-min lung volumes with both laryngeal mask airway and combitube were significantly higher (laryngeal mask airway 15.0+/-6.61, combitube 16.6 +/- 6.81 vs bag-valve-mask 4.8 +/- 2.71, P < 0.01). The time for insertion was significantly faster with both bag-valve-mask and laryngeal mask airway compared with the combitube (median: bag valve mask 22 s, laryngeal mask airway 37 s vs combitube 70 s, P < 0.01). This may tip the scales towards using the laryngeal mask airway during basic life support airway management. In conclusion, our data suggests that both laryngeal mask airway and combitube may be appropriate alternatives for airway management in the first few minutes of CPR. PMID- 10459595 TI - Cell death, calcium mobilization, and immunostaining for phosphorylated eukaryotic initiation factor 2-alpha (eIF2alpha) in neuronally differentiated NB 104 cells: arachidonate and radical-mediated injury mechanisms. AB - These experiments examine the effects of arachidonate with respect to cell death, radical-mediated injury, Ca2+ mobilization, and formation of ser-51 phosphorylated eukaryotic initiation factor 2alpha [eIF2alpha(P)]. It is known that during brain ischemia the concentration of free arachidonate can reach 180 microM, and during reperfusion oxidative metabolism of arachidonate leads to generation of superoxide that can reduce stored ferric iron and promote lipid peroxidation. During early brain reperfusion, we have shown an approximately 20 fold increase in eIF2alpha(P) which maps to vulnerable neurons that display inhibition of protein synthesis. Here in neuronally differentiated NB-104 cells, equivalent cell death (assessed by LDH release) was induced by 40 microM arachidonate and 20 microM cumene hydroperoxide (CumOOH, a known alkoxyl radical generator). In these injury models (1) radical inhibitors (BHA, BHT, and the lipophilic iron chelator EMHP) block CumOOH-induced cell death but do not block arachidonate-induced death; (2) 40 microM arachidonate (but not up to 40 microM CumOOH) rapidly induces Ca2+ release from intracellular stores; (3) both 40 microM arachidonate and 20 microM CumOOH induce intense immunostaining for eIF2alpha(P); and (4) the elF2alpha(P) immunostaining induced by CumOOH but not that induced by arachidonate is completely blocked by anti-radical intervention with EMHP. Arachidonate-induced formation of eIF2alpha(P) and cell death do not require iron-mediated radical mechanisms and are associated with Ca2+ release from intracellular stores; however, radical-mediated injury also induces both eIF2alpha(P) and cell death without release of intracellular Ca2+. Our data link eIF2alpha(P) formation during brain reperfusion to two established injury mechanisms that may operate concurrently. PMID- 10459596 TI - Lazarus phenomenon: another case? PMID- 10459597 TI - Presidential Address. From whence we cometh. PMID- 10459598 TI - Minimally invasive plate osteosynthesis: does percutaneous plating disrupt femoral blood supply less than the traditional technique? AB - OBJECTIVES/HYPOTHESIS: Proximal and distal femur fractures have traditionally been treated with open reduction and internal fixation through a standard lateral approach. New, "minimally invasive" internal fixation techniques, however, have been developed in an effort to devascularize the bone less than the traditional method. The purpose of this study was to determine whether a minimally invasive percutaneous plating technique better preserves bone vascularity relative to the traditional method by comparing the effect of the two approaches on the blood supply of the distal femur using silicone arterial dye injection in a cadaveric model. STUDY DESIGN/METHODS: Ten fresh human cadavers underwent lateral conventional plate osteosynthesis (CPO) through a standard lateral approach on one side and minimally invasive plate osteosynthesis (MIPO) through two three centimeter incisions on the contralateral side. After injection of silicone dye, a dissection was performed bilaterally to identify the femoral perforating and nutrient arteries. RESULTS: All MIPO specimens showed intact perforating and nutrient arteries, whereas the CPO specimens had a variable incidence of vessel disruption. The MIPO group demonstrated better periosteal perfusion in each of the cadavers and improved medullary perfusion in 70 percent of the MIPO specimens compared with the CPO specimens. CONCLUSION: A percutaneous minimally invasive plating technique disrupts the femoral blood supply less than the traditional open method. Such minimally invasive methods may be more advantageous biologically than the traditional method. PMID- 10459599 TI - Biomechanical evaluation of impaction fractures of the femoral head. AB - OBJECTIVES: To measure the effect of an impaction fracture of the femoral head on load transmission in the hip joint. DESIGN: We measured the contact areas and pressure between the acetabulum and femoral head of cadaveric pelves in four different conditions: intact, with an operatively created one-square-centimeter defect in the superior femoral head, with a two-square-centimeter defect, and with a four-square-centimeter defect. All defects were uniformly three millimeters deep. SETTING: Hips were loaded in a simulated single-limb stance. Pressure and area measurements were made with Fuji pressure-sensitive film. SPECIMENS: Seven hip joints in seven whole pelves were tested. MAIN OUTCOME MEASUREMENTS: Contact area, load, and mean and maximum pressures were measured. RESULTS: Peripheral loading was seen in the intact acetabulum. This was not disrupted after impaction fractures of any size. A significant increase in mean maximum pressures in the superior acetabulum was seen with two-square-centimeter and four-square-centimeter defects. CONCLUSIONS: In contrast to prior biomechanical studies of acetabular fractures, our investigation revealed that disruption of the peripheral distribution of load does not occur with impaction fractures of the femoral head. Clinical series indicate that impaction injuries to the femoral head are associated with a poor prognosis. Previous biomechanical data on acetabular fracture patterns associated with a poor prognosis have shown increases in mean and peak pressures in the superior acetabulum. This was seen with two-square-centimeter and four-square-centimeter impaction injuries. Other factors, such as wear of the articular cartilage during joint motion or associated microscopic damage to the remainder of the joint surface at the time of injury, may also contribute to the rapid joint deterioration seen in these injuries. Further study is indicated. PMID- 10459600 TI - Intraoperative fluoroscopy to evaluate fracture reduction and hardware placement during acetabular surgery. AB - OBJECTIVES: To evaluate use of intraoperative fluoroscopy during acetabular surgery to determine fracture reduction and accurate placement of screws. DESIGN: Retrospective. SETTING: Level I trauma center. PARTICIPANTS: Thirty patients with thirty-two acetabular fractures. INTERVENTION: Patients were evaluated with fluoroscopy during surgery to assess fracture reduction and screw placement. Anterior-posterior (AP), oblique, and lateral pelvic fluoroscopic images were obtained intraoperatively. Postoperative radiographs were used to verify fluoroscopic findings; computed tomography (CT) scans were used as the control to assess intraarticular screw placement. MAIN OUTCOME MEASUREMENTS: Radiographic and clinical assessment of fracture reduction and screw placement. RESULTS: Intraoperative fluoroscopy confirmed the extra-articular position of all screws evaluated. Postoperative CT scans confirmed the extra-articular placement of all screws assessed by fluoroscopy. Quality of reduction using intraoperative fluoroscopic images had a 100 percent correlation with reduction on final radiographs. One patient, with two screws placed without fluoroscopic evaluation, had intra-articular placement requiring revision surgery. CONCLUSIONS: Intraoperative fluoroscopy is effective in evaluating both acetabular fracture reduction and hardware placement. PMID- 10459601 TI - Comparative biomechanics of hybrid external fixation. AB - OBJECTIVE: To compare the elastic stiffness, in several loading modes, of commercially available hybrid external fixation systems. DESIGN: Laboratory investigation using a polyvinylchloride pipe periarticular tibia fracture model. SETTING: Simulated periarticular fractures were created in an in vitro tibia fracture model. Instrumented specimens and intact controls were elastically tested in a biomaterials testing system. INTERVENTION: Groups of simulated periarticular tibia fractures were stabilized with one of six different hybrid external fixator designs. MAIN OUTCOME MEASUREMENTS: Elastic stiffness of each specimen was measured in compression, torsion, flexion bending, extension bending, and varus/valgus bending. RESULTS: Fixators with multiple levels of fixation in the periarticular fragment, regardless of design, were stiffer than those with one level. Specifically, the EBI Ring Connector fixator was stiffer than all others in all modes of testing. The Ace, Synthes, Smith & Nephew Richards, and How medica fixators were mechanically similar. The Zimmer Torus fixator was the least stiff fixator tested. CONCLUSIONS: Fixators with multiple levels of fixation in the periarticular fragment, regardless of design, were stiffer than those with one level. The choice of which hybrid external fixator to use should be made based not only on stiffness but also on ease of clinical application, patient comfort, customer support from the manufacturer, and cost. Clinical investigation of the efficacy of each of these devices is warranted. PMID- 10459602 TI - Closed reduction/percutaneous fixation of tibial plateau fractures: arthroscopic versus fluoroscopic control of reduction. AB - OBJECTIVE: To evaluate arthroscopic versus fluoroscopic reduction and percutaneous fixation of lateral tibial plateau fractures of AO/OTA Types 41.B1 to 41.B3. DESIGN: Prospective study. SETTING: University hospital. PATIENTS AND INTERVENTION: One hundred sixty-eight patients underwent operative treatment for a tibial plateau fracture from 1988 to 1995. Thirty-three of these patients had monocondylar fractures of the lateral plateau that were treated by percutaneous reduction and fixation techniques. In the first ten cases, arthroscopic control of reduction was used. The following twenty-three consecutive cases were treated by reduction and fixation solely under fluoroscopic control. The arthroscopy group was followed for a mean of fifty-two months and the fluoroscopy group for thirty-eight months. RESULTS: Nine of ten cases of the arthroscopy group had an excellent or good result in Rasmussen's knee score at follow-up. One patient with an unreduced anterolateral depression zone despite arthroscopic surgery required a total knee prosthesis after eighteen months. Sixteen cases in the fluoroscopy group met the follow-up criteria. Fifteen were graded good or excellent in Rasmussen's clinical score; sixteen were excellent or good in the radiological score. One patient claimed chronic medial joint line pain after a lateral split fracture and had arthroscopy revealing chondral degeneration on the medial side but had no pathological findings in the lateral compartment. No secondary meniscus or ligament surgery was performed in the follow-up period. CONCLUSIONS: Percutaneous treatment of fractures of the tibial plateau can be performed using arthroscopy as well as image intensification to control reduction of the joint surface. We were not able to demonstrate any significant benefit from arthroscopy compared with fluoroscopic reduction. Reduction under image intensification is technically easier in our practice, especially in serial fractures and multiply injured patients. We reserve arthroscopy for cases with significant ligament injuries and for children with fractures of the median eminence. PMID- 10459603 TI - Orthopaedic trauma education: visions for the future through the OTA. Orthopaedic Trauma Association. AB - As the trauma center system continues to expand, not only will the requirement for more and better trained trauma surgeons increase, but the means of educating them will need to become more standardized. The general surgeons recognized this many years ago, but orthopaedic trauma has lagged in its efforts to present a coordinated academic and clinical program to residents and fellows. The Orthopaedic Trauma Association has made a move to develop guidelines which may be used by training programs in an effort to improve the educational standards of this subspecialty. The recruitment and retention of young orthopaedic trauma surgeons remains an issue. PMID- 10459604 TI - Guidelines for orthopaedic trauma fellowships. Orthopaedic Trauma Association, Fellowship and Career Choices Committee. PMID- 10459605 TI - Resident education curriculum for orthopaedic trauma. Orthopaedic Trauma Association, Fellowship and Career Choices Committee. PMID- 10459606 TI - Nonunion of a pediatric lateral condyle fracture without ulnar nerve palsy: sixty year follow-up. AB - Displaced lateral condyle fractures in the pediatric population are usually treated with open reduction and internal fixation. Significant complications associated with the nonoperative management include nonunion, malunion, deformity, and tardy ulnar nerve palsy. However, few cases of nonunion of the lateral condyle and tardy ulnar nerve palsy with long-term follow-up have been reported. We present a radiographically documented case of a pediatric lateral condyle fracture and subsequent nonunion with significant cubitus valgus deformity without ulnar nerve palsy sixty years following injury. PMID- 10459607 TI - Pediatric elbow dislocation associated with a milch type I lateral condyle fracture of the humerus. AB - A Milch Type I lateral condyle fracture associated with a posterior elbow dislocation is described in a pediatric patient. Previously, Milch Type I fractures were thought to be stable injuries due to maintenance of the lateral trochlear rim. Prompt recognition and treatment are essential to avoid complications of this injury and to ensure a good functional result. PMID- 10459608 TI - The East Baltimore Lift: a simple and effective method for reduction of posterior hip dislocations. PMID- 10459610 TI - The full-field flicker test in glaucomas: influence of intraocular pressure and pattern of visual field losses. AB - BACKGROUND: The purpose of this study was to evaluate how temporal contrast sensitivity (TCS) determined with full-field flicker stimuli is influenced by intraocular pressure and whether TCS is reduced in glaucoma patients with diffuse perimetric losses as well as in patients with localized visual field deficits. METHODS: TCS was determined with sinusoidally flickering light (37.1 Hz) in a full-field bowl. Perimetric mean defect (MD) and cumulative defect curves (Octopus G1) were used to distinguish between patients with localized and diffuse field deficits. Normal subjects (296), low-tension glaucoma patients (98) and open-angle glaucoma patients with previously elevated intraocular pressure (541) were classified into five subgroups taking into account the depth of their visual field losses. RESULTS: No significant correlation between full-field flicker sensitivity and prevailing intraocular pressure was found in normals (Y=1.36+0.006 X) or in patients (Y=0.95-0.0002 X). Analyses of validity at a predefined specificity of 90% reveal a reduction of TCS in patients with early (MD<5 dB) diffuse perimetric losses (sensitivity 69%) as well as in those showing localized visual field defects (sensitivity 65%). Sensitivity was 87% in patients with diffuse perimetric defects (MD 5-10 dB), 93% in a group of patients with both types of losses, and 100% in advanced glaucomas (MD>20 dB). The lack of TCS is similar in open-angle glaucomas and in field-loss-matched normal-tension glaucoma patients. CONCLUSIONS: Significantly reduced TCS in patients with early diffuse perimetric losses as well as in those showing localized visual field defects indicates that localized damages can be associated with general deterioration of the ability to perceive flickering stimuli. Thus, this flicker test can be performed in a full-field bowl with no need for fixation. Considering its other clinical qualities (photopic conditions, low influence of prevailing intraocular pressure and media opacity) the test may be a useful, convenient supplementary procedure in glaucoma screening. PMID- 10459609 TI - Diagnosis of chlamydial infection by direct enzyme-linked immunoassay and polymerase chain reaction in patients with acute follicular conjunctivitis. AB - BACKGROUND: Acute follicular conjunctivitis is a clinical diagnosis common to multiple etiologies, of which chlamydial infection requires specific antibiotic treatment. PURPOSE: This prospective study was designed to evaluate Chlamydia trachomatis as the cause of acute follicular conjunctivitis by two sensitive tests: direct enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). METHODS: Conjunctival scrapings from patients presented with untreated acute follicular conjunctivitis were examined by ELISA and PCR, and patients were followed up for prolongation of the disease course. RESULTS: All 36 consecutive patients presented with acute follicular conjunctivitis were negative for Chlamydia trachomatis by ELISA and PCR. None of the patients had a prolonged course of more than 4 weeks or required treatment with systemic antibiotics as would be expected from chlamydial infection. CONCLUSIONS: Chlamydia trachomatis was probably not responsible for the acute follicular conjunctivitis in this series, and ELISA and PCR may not be cost effective for evaluation of acute follicular conjunctivitis due to chlamydial infection. Further evaluation of the cost effectiveness of these tests is required in chronic follicular conjunctivitis. PMID- 10459611 TI - Predictive value of pattern VEP, pattern ERG and hole size in macular hole surgery. AB - OBJECTIVE: To describe pattern-reversal visual evoked response (PRVEP) and pattern electroretinogram (PERG) parameters in eyes with macular hole and their value for predicting postoperative visual outcome. METHODS: Prospectively we studied 27 eyes (27 patients) with a full-thickness macular hole. Preoperatively the hole and rim were measured and the PRVEP and PERG were recorded. The preoperative parameters were correlated with postoperative visual outcome. RESULTS: The macular hole was closed in 26 of 27 eyes. Sixteen eyes (59%) had an increase in visual acuity (VA) of two lines or more, 10 eyes (37%) remained within one line of preoperative VA and 1 eye (4%) had a decrease in VA of two lines. Duration of symptoms was negatively correlated with preoperative VA (R= 0.547, P=0.0038) and postoperative VA (R=-0.519, P=0.0065) and positively correlated with hole area (R=0.533, P=0.0061) and rim area R=0.633, P=0.0009). Only the PRVEP P100 latency of the 10' check size and the PERG N35 latency were significantly associated with visual outcome (P=0.022 and P=0.042 respectively). CONCLUSIONS: There was no association of either hole or rim size with postoperative visual outcome. Preoperative electrophysiology, however, is useful as a prognostic tool. Utilization is limited to the use of latency parameters of the response and is dependent on the check size of the stimulus. PMID- 10459612 TI - A temporal deficit in juvenile diabetics. AB - BACKGROUND: In this study we examined the temporal domain of visual function in diabetics without retinopathy by examining wavelength discrimination ability at two exposure durations. The results were compared to those found by heterochromatic brightness matching and anomaloscope matches. METHODS: Wavelength discrimination was performed between 440 and 540 nm at exposure times of 1 s and 0.04 s in eight juvenile diabetic patients without retinopathy. The monochromatic stimuli were presented in Maxwellian view and were set to be equally bright prior to the experiment using heterochromatic brightness matching. In addition, Rayleigh and Moreland anomaloscope matches were performed. The results of the diabetic group were compared to those of an age-matched control group of eight subjects with normal colour vision. RESULTS: Wavelength discrimination showed no difference between the groups for an exposure time of 1 s. With an exposure duration of 0.04 s, however, the diabetics show raised thresholds for the shortest wavelengths tested. In addition, brightness matches were increased at the short wavelengths, and anomaloscope matches showed a decrease in the match range for the Moreland (blue-yellow) equation. CONCLUSION: The results indicate post-receptoral alterations in diabetic patients with no visible changes in their retinae. PMID- 10459613 TI - Relationship of the distal optic nerve sheath to the circle of Zinn. AB - BACKGROUND: This study was designed to determine the relationship of the vascular circle of Zinn (CZ) and its branches to the termination of the subarachnoid space surrounding the optic nerve sheath, with particular reference to optic disc size. METHODS: Serial sections of 29 normal human optic nerves were performed at 6 microm intervals. The position of the CZ and subarachnoid space were measured with WILD Heerbrugg objective graticules. The results were analysed with reference to the optic disc size. RESULTS: The position of the CZ was variable but two major types were recognised. In type 1 the circle was located anterior to the distal sheath, and in type 2 posterior to the sheath. There were more small optic discs observed in type 2. CONCLUSION: In this study the CZ was more posteriorly located in small optic discs. This location may induce risk factors contributing to the development of ischaemic optic neuropathy. The combination of small discs, posterior placement of the CZ, and anatomical variations in the vascular pattern may predispose to ischaemic events. PMID- 10459614 TI - The residual epiretinal membrane after vitrectomy for macular hole. AB - BACKGROUND: We retrospectively observed idiopathic macular holes in 63 eyes using a scanning laser ophthalmoscope, in order to study the relation between postoperative epiretinal membranes and closure of macular holes following vitrectomy. METHODS: The eyes were classified into three groups based on the degree of the postoperative epiretinal membranes. Group I consisted of 23 eyes with no epiretinal membrane remaining on the retina after vitrectomy. Group II consisted of 20 eyes in which epiretinal membranes were observed on the retina, but separate from the edge of the macular hole. Group III consisted of 20 eyes in which epiretinal membranes were observed at the edge of the macular hole. Using these three groups, we studied how postoperative epiretinal membranes were related to the closure of macular holes. RESULTS: All macular holes (100%) in groups I and II were closed following vitrectomy. In group III, 5 (25.0%) of 20 eyes had complete closure and 13 eyes (65.0%) had incomplete closure of the macular hole, while 2 eyes (10.0%) had re-opening of initially closed macular holes several months after vitrectomy. CONCLUSION: Residual postoperative epiretinal membranes at the edge of macular hole are responsible for primary failure of vitrectomy. Removal of epiretinal tissues around the macular hole is important for macular hole to be closed following vitrectomy. PMID- 10459615 TI - Histopathological changes in iridocorneal angle of inherited glaucoma in rabbits. AB - BACKGROUND: We examined morphologically the angular region of eyes affected by inherited glaucoma in rabbits genetically developed by crossbreeding in order to investigate the etiologic changes in the iridocorneal angle and to establish whether this strain of rabbit is a suitable animal model of goniodysgenetic glaucoma in humans. METHODS: The angular regions of both normal and glaucomatous eyes from four rabbits having unilateral inherited glaucoma were observed with light and electron microscopy. RESULTS: In the glaucomatous eyes angular region, the aqueous plexus corresponding to Schlemm's canal in humans was open and located far peripherally to the peripheral margin of the anterior chamber angle, although the plexus of one glaucomatous eye was poorly developed with a small lumen. In the angular meshwork, which corresponds to the trabecular meshwork in humans, a thick abnormal tissue with round cells embedded in the extracellular matrix was located just beneath the plexus. A large amount of extracellular matrix of basal lamina-like material was observed in the thick tissue. In the normal eyes, the angular region consisted of well-developed trabecular sheets with neither a thick tissue nor accumulations of extracellular matrix in the angular meshwork. CONCLUSION: The findings observed in the glaucomatous eyes are much the same as those observed in goniodysgenetic glaucoma in humans, suggesting that this strain of inherited glaucoma rabbits is a suitable animal model of goniodysgenetic glaucoma in humans. The present study also supports the hypothesis that the presence of a thick subcanalicular tissue due to maldevelopment of the iridocorneal angle is one of the main causes of this type of glaucoma. PMID- 10459616 TI - Effects of beta antagonists on mechanical properties in rabbit ciliary artery. AB - BACKGROUND: We set out to clarify the mechanisms involved in the effectiveness of beta-adrenergic receptor antagonists on the ocular circulation. METHODS: The effects of three beta antagonists, timolol, betaxolol and carteolol, on the isolated rabbit ciliary artery were investigated in vitro using isometric tension recording methods. RESULTS: Phenylephrine dose-dependently contracted ciliary artery smooth muscle, and bunazosin (1 microM) shifted this dose-response curve to the right. Isoproterenol, on the other hand, had no effect up to the concentration of 1 mM. Betaxolol and timolol had no effect on the ciliary artery. However, carteolol dose-dependently contracted this muscle from a concentration of 1 microM. After precontraction by excess-[K]0 solutions, application of betaxolol or timolol dose-dependently provoked relaxation; the minimum concentration of betaxolol or timolol required to generate the relaxation was 100 microM and 300 microM, respectively. Carteolol did not generate relaxation at concentrations up to 1 mM. After pretreatment with L-NAME (300 microM), the amplitude of relaxation induced by 10 microM carbachol was reduced to 33.0+/ 20.2%, while betaxolol- or timolol-induced relaxation was unchanged. Diltiazem (10 microM) induced relaxation which was not inhibited by pretreatment with L NAME. CONCLUSION: Betaxolol and timolol could directly relax rabbit ciliary artery in vitro at relatively high concentrations, and relaxation was not due to NO released from the preparation. Presumably, this relaxation occurs through action similar to Ca antagonists. However, the clinical importance of this effect is not yet clear. Carteolol had no relaxant effect in vitro. PMID- 10459617 TI - Surgically induced degeneration and regeneration of the choriocapillaris in rabbit. AB - BACKGROUND: Retinal pigment epithelium (RPE) and choriocapillaris play an important role in maintaining the outer retina. Clinical and experimental studies have shown that there is a close relationship between them. To examine the relationship between RPE and choriocapillaris we developed an animal model in which degeneration and regeneration of the choriocapillaris can be created easily and reproducibly. METHODS: Using pigmented rabbits a retinal detachment (about 7 disk diameters) was created in the eye and the detached retina was surgically removed. A half area of the exposed RPE was mechanically debrided using a silicone brush. The other half area remained untouched. The eyes were morphologically examined at 3, 7, and 14 days and 4 and 8 weeks after the surgery using scanning or transmission electron microscopy. Changes in the choriocapillaris after the RPE debridement were also evaluated by corrosion vascular casts. RESULTS: The debrided area was resurfaced with elongated RPE in 7 days after surgery. The replaced RPE was dedifferentiated but it gradually recovered cell polarity by 4 weeks after surgery. The choriocapillaris beneath the debrided area underwent degeneration; however, it regenerated by 4 weeks after surgery, corresponding to the timing of the morphologic recovery of the replaced RPE. CONCLUSION: This animal model of surgically induced degeneration and regeneration of the choriocapillaris may be useful to clarify the relationship between RPE and choriocapillaris and to study potential treatments for choroidal vascular diseases. PMID- 10459618 TI - Transfection of basic fibroblast growth factor (bFGF) gene or bFGF antisense fene into human retinal pigment epithelial cells. AB - BACKGROUND: Transplantation of RPE cells offers a potential of restoring retinal pigment epithelium (RPE) function and has been shown to be effective in the dystrophic RCS rat model. Recently, RPE transplantation was attempted in patients with age-related macular degeneration. Basic fibroblast growth factor (bFGF) plays important roles in maintaining normal retinal function. The purpose of this study was to introduce bFGF sense or antisense cDNA into human RPE cells to alter the expression of bFGF. METHODS: Human bFGF sense cDNA or antisense cDNA was inserted into the pBK-CMV vector. For stable gene expression, we introduced the plasmids into RPE cells using the electroporation method. Following electroporation, transfected RPE cells were cultured and resistant cells were selected in the presence of antibiotic G418. We analyzed the expression of the transfected genes in the cloned RPE cells by polymerase chain reaction (PCR) and by reverse transcription (RT)-PCR. RESULTS: Cloned RPE cells in which the bFGF sense or antisense cDNA had been efficiently transfected were established. PCR and RT-PCR analysis demonstrated not only the presence but also the expression of bFGF sense or antisense cDNA in the transfected RPE cells. CONCLUSIONS: Human bFGF sense cDNA or antisense cDNA can be efficiently introduced into cultured RPE cells by the electroporation method. The successful expression of the genes into RPE cells demonstrated that this technique can be used to regulate bFGF expression and thus increase the scope of RPE transplantation for the treatment of retinal diseases. PMID- 10459619 TI - Ultrastructural localization of lipid peroxides as benzidine-reactive substances in the albino mouse eye. AB - BACKGROUND: Lipid peroxidation is considered to be a prominent feature of retinal degeneration and has also been proposed to be involved in the pathogenesis of age related macular degeneration. Melanin protects against lipid peroxidation and takes part in the detoxification of lipid peroxides (LP). LP can be ultrastructurally detected as benzidine-reactive substances (BRS) using tetramethylbenzidine (TMB). Albino mice lack melanin. In the present study, LP were localized as BRS in the eyes of albino and pigmented mice. METHODS: Eye cups of an albino mouse lineage and of wild-type mice were fixed with 2% glutaraldehyde, incubated with 0.5 mg/ml TMB and embedded for electron microscopy. RESULTS: BRS were detected in the eyes of albino mice, but no reaction product was seen in pigmented eyes. BRS located in the retinal pigment epithelium (RPE) and in the choroid of the albino mouse; no BRS were found in intact rod outer segments (ROS). CONCLUSION: The lack of melanin in albino mice is associated with a higher level of lipid peroxidation in RPE and choroid. Melanin seems to protect against LP in RPE and choroid. A lack of melanin is not associated with lipid peroxidation in intact ROS. The present investigation demonstrates the significance of melanin in protection against LP in RPE and choroid. PMID- 10459620 TI - The effect of tranilast on experimental proliferative vitreoretinopathy. AB - BACKGROUND: Tranilast has been clinically used for various allergic diseases. Recently, it has also been found to inhibit excessive scarring in wound healing processes. In this study, we examined the effects of tranilast on the treatment for experimental proliferative vitreoretinopathy (PVR). METHODS: Cultured rabbit conjunctival fibroblasts were injected intravitreously (50000 cells/eye) into the rabbit vitreous to induce experimental PVR. Immediately after that, tranilast (0.5-5 mg/ml, 0.1 ml/eye) was injected into the vitreous. Injection of vehicle solution was used as a negative control. PVR was clinically evaluated by masked observers using ophthalmoscopy and graded into six stages: 0 (no PVR) to 5 (severe PVR). The amount of transforming growth factor beta1 (TGF-beta1) in the vitreous was measured by ELISA method. Functional and morphological changes induced by 5 mg/ml tranilast were sought by electroretinography, light microscopy, and electron microscopy on day 28. RESULTS: The average stage of PVR in the eyes treated with tranilast (1 or 5 mg/ml) was significantly lower than that in the control group on days 14 and 28. There was no difference between the eyes treated with low-dose tranilast (0.5 mg/ml) and the control group. The amount of TGF-beta1 in the vitreous of tranilast-treated eyes was significantly lower than in the control group. The morphological and functional studies did not show any deleterious effect of tranilast on the retinal function and morphology. CONCLUSION: Tranilast effectively inhibits the progression of PVR without showing apparent toxicity of the eye. This agent has therapeutic value for PVR. PMID- 10459621 TI - Confocal microscopy in lattice corneal dystrophy. AB - BACKGROUND: The purpose of the study was to assess the appearance of lattice corneal dystrophy by means of white-light confocal microscopy. METHODS: Two consecutive patients with lattice corneal dystrophy were prospectively examined. In vivo white-light tandem-scanning confocal microscopy was performed in the right eye of the first patient. Her left eye had undergone penetrating keratoplasty 4 years earlier. Histologic findings of the corneal button were compared with confocal microscopic findings of the right eye. The other patient was monocular and confocal microscopy was performed only in the non-seeing eye. RESULTS: In both patients, linear and branching structures with changing reflectivity and poorly demarcated margins were visualized in the stroma. The linear structures measured approximately 40-80 microm in width. CONCLUSION: Lattice corneal dystrophy presents characteristic linear images on confocal microscopy and should not be misdiagnosed as fungal hyphae in cases of corneal infection. PMID- 10459622 TI - Nonalcoholic steatohepatitis (NASH): further expansion of this clinical entity? PMID- 10459623 TI - Strategy of liver-directed gene therapy: present status and future prospects. AB - The liver is particularly amenable to gene therapy as it is the site of many metabolic diseases and malignancies. Thus, liver-directed gene therapy is being actively pursued and developed as a method of treatment for various liver diseases. Strategies of liver-directed gene therapy include drug delivery to the liver, compensation of the defective gene(s), anti-tumor activity, anti-viral therapy, and immunomodulation. The strategy chosen for liver-directed gene therapy depends on the genetic basis of the disease. Many aspects are key factors to the success of the chosen strategy: intervention of genes, efficient gene delivery system, stable transgene expression, transgene regulation, target cell transfection, and timing of transgene expression. Several tactics can be used to overcome problems in the above, and these include the use of a gene switch to exogenously regulate transgene expression, targeting at the transcriptional level, circumvention of the immune response (as in the use of adenovirus vector to achieve long-term correction of genetic diseases), and genetically engineered antibodies in gene transfer. At the present rate of research activity and development, gene therapies may soon be more efficient than current standard treatments for some liver diseases. PMID- 10459624 TI - Interferon versus ribavirin plus interferon in chronic hepatitis C previously resistant to interferon: a randomized trial. AB - BACKGROUND: More than 70% of patients with chronic hepatitis C are resistant to interferon therapy. Ribavirin, in association with interferon, has been demonstrated as effective, at a dose of 800-1200 mg/day, but the efficacy of a lower dose has not been established. METHODS: We assessed the effectiveness of the combination of 600 mg/day of ribavirin plus 3 MU of interferon over a period of 6 months, in a group of patients previously resistant to interferon. Sixty-two patients with chronic hepatitis C with serum and hepatic HCV RNA relapsers or non responders to interferon, were randomly divided into two groups: group A received 3 MU of interferon alpha-2b, three times a week for 6 months; group B was given the same dose plus 600 mg per day of ribavirin for 6 months. Two patients from each group dropped from therapy. One patient from group A and two from group B withdrew from treatment because of adverse effects. RESULTS: Mean alanine aminotransferase levels were similar in both groups throughout the study. A sustained response was observed in 7% and 7.4% of groups A and B with short-term response in 39% and 59%, and no response in 54% and 34% from both groups respectively (non-significant). At 12 months, 4 and 7 patients from groups A and B respectively, cleared serum HCV RNA however, only one sustained responder from each group cleared HCV RNA from peripheral blood mononuclear cells. At 18 months, 3 patients remained serum HCV RNA negative. Adverse effects were similar. Only haemoglobin values were lower in group B in the first month of therapy (p<0.05). CONCLUSION: In conclusion, the combination of 3 MU of interferon plus 600 mg of ribavirin is not effective in chronic hepatitis C resistant to interferon. PMID- 10459625 TI - Frequent expression of mucin core protein MUC1 in non-neoplastic gallbladder mucosa from patients with pancreaticobiliary maljunction. AB - BACKGROUND: Gallbladder carcinoma is known to develop frequently in patients with pancreaticobiliary maljunction, though the causal relationship remains speculative. METHODS: Histopathologic changes, expression of mucin core protein MUC1 and MUC2, and cell proliferative activities in the gallbladder mucosa from 27 patients with panceaticobiliary maljunction and 21 control gallbladders were examined. Three cases of pancreaticobiliary maljunction were associated with gallbladder carcinoma. RESULTS: The lining epithelia of the non-neoplastic gallbladder mucosa of pancreaticobiliary maljunction showed frequently papillary hyperplasia and higher proliferative activities, when compared to the control. In 3 cases with carcinoma, MUC1 was expressed on the luminal border and in the cytoplasm of carcinoma cells, particularly in de-differentiated and invasive areas. MUC1 was variably expressed on the luminal surface of the lining epithelia of non-neoplastic gallbladder mucosa in babies, children, youths and adults with pancreaticobiliary maljunction. However, such expression was focally seen in 2 of the 21 control cases (p<0.01). MUC2 was scattered in the hyperplastic and carcinomatous epithelial cells appearing as goblet cells in pancreaticobiliary maljunction and control groups. CONCLUSIONS: This study suggests that persistent MUC1 expression and increased cell proliferative activities of non-neoplastic gallbladder epithelium of the patients with pancreaticobiliary maljunction after birth reflect an altered phenotype of epithelial cells and these abnormalities may be related to carcinogenesis in such patients. PMID- 10459626 TI - Relationship of hepatitis C viremia to HIV state and to infection by specific hepatitis C genotypes. AB - AIMS/BACKGROUND: This study was undertaken to examine the relationship of hepatitis C (HCV) viremia to human immunodeficiency virus (HIV) infection and to investigate the evidence of infection by specific hepatitis C genotypes. PATIENTS AND METHODS: The analysis of HCV viremia was performed by reverse transcription polymerase chain reaction in serum samples from 186 patients' selected by their positivity for anti-HCV antibodies. All samples were tested for HIV infection. In those patients with sera positivity for HCV RNA, isolates were genotyped by line probe assay. In those patients whose sera were negative for HCV RNA, antibodies specific for genotypes implicated in past HCV infection were detected by ELISA. RESULTS: HCV RNA was detected in 117 patients with anti-HCV antibodies (62.9%). There was no statistically significant association between HCV RNA and HIV positivity (Odds ratio, O.R.: 1.75, Confidence interval 95%, C.I.: 0.92-3.33, p = 0.095). A positive association was demonstrated between infection by HCV genotype 3 and HCV viremia (O.R.: 10.67, C.I.: 1.51-458.05, p = 0.015) in HIV-infected patients, as well as between infection by HCV genotype 1 and HCV viremia (O.R.: 4.71, C.I.: 1.65-13.75, p = 0.002) in HIV-non infected individuals. In both groups, a negative association was observed between past HCV infection by multiple genotypes and HCV viremia (HIV-infected patients, O.R.: 0.10, C.I.: 0.00 1.11, p = 0.033. HIV-non infected patients, O.R.: 0.05, C.I.: 0.00-0.41, p = 0.001). CONCLUSIONS: Infection by specific HCV genotypes (type 3 in HIV-infected patients and by type 1 in HIV-non infected ones) implies a higher risk of HCV viremia, whereas multiple HCV types infection is negatively associated with this probability. HIV coinfection does not influence the probability of HCV viremia. PMID- 10459627 TI - HBeAg immunostaining of liver tissue in various stages of chronic hepatitis B. AB - AIMS: We studied the tissue expression of hepatitis B e antigen (HBeAg) in 29 liver biopsies from 27 HBV carriers. METHODS: HBeAg expression was assessed in relation to HBeAg in serum, precore mutations, HBV DNA levels and liver damage as measured by histology activity index. RESULTS: HBeAg in liver tissue was detected by immunostaining in 6 of 7 patients positive for HBeAg in serum. In patients negative for HBeAg in serum, HBeAg was detected in none of 11 specimens from patients infected exclusively with a precore mutant that disrupts HBeAg synthesis, as compared with 3 of 11 specimens from patients carrying HBV with an intact precore region. These 3 patients all showed high HBV DNA levels in serum and severe liver damage. CONCLUSIONS: Overall, viral replication was strongly associated with the cytoplasmic HBeAg and nuclear HBcAg staining, but not with tissue staining for HBsAg. Because of the close relationship between tissue HBeAg expression and high viral load, the pathogenetic importance of HBeAg remains unclear. PMID- 10459628 TI - Nonalcoholic steatohepatitis: a toxic liver disease in industrial workers. AB - AIMS: Occupational/environmental exposure to hepatotoxins has recently been implicated in nonalcoholic steatohepatitis (NASH). The aims of this study were to determine the presence and frequency of NASH in a large group of workers chronically exposed to several volatile petrochemical products in an industrial area in north-east Brazil and to observe its course in workers removed from the work environment. METHODS: 1500 asymptomatic workers were screened with standard liver blood tests during 1994-5. Those with elevated transaminases (>3x normal) on 3 occasions were evaluated further both clinically and with serum HBsAg, anti HCV, ferritin, lipids and autoantibody determination. Patients with either no etiological diagnosis, positive HBsAg/anti-HCV serology and/or excess alcohol intake underwent liver biopsy. Those with obesity, diabetes or an isolated abnormal GGT were excluded. Of workers diagnosed as having NASH (compatible histology and no excess alcohol intake), a proportion were removed from the work environment and evaluated monthly with liver blood tests and a repeat liver biopsy 8-14 months later. RESULTS: 112 workers had abnormal transaminases and 32 fulfilled the criteria for liver biopsy. 20 of these were classified as NASH, the remainder had viral hepatitis (n = 6), alcoholic liver disease (n = 5) or portal vein thrombosis (n = 1). In all of the 10/20 who were removed from the work environment, their aminotransferases and GGT gradually decreased and their histology improved. CONCLUSIONS: These results demonstrate that NASH can occur following chronic exposure to volatile petrochemical substances in the workplace. Exposed workers should be regularly screened for the presence of liver damage and ideally removed from the work environment where possible. PMID- 10459629 TI - Loss of heterozygosity and microsatellite instability in human non-neoplastic hepatic lesions. AB - AIMS/BACKGROUND: Carcinogenesis is thought to be a multistage process that occurs as a result of mutations in oncogenes and tumor suppressor genes. One way to monitor a vast range of these changes is by microsatellite PCR amplification that detects loss of heterozygosity and microsatellite instability between normal and tumor specimens of the same subject. Viral cirrhosis is considered a strong predisposing factor for the development of liver cancer. The aim of the study therefore was to examine precancerous hepatic lesions and compare them with others not considered as high risk for hepatocellular carcinoma. METHODS: We examined 43 subjects for 19 microsatellite markers spanning chromosomes 1, 9 and 17. Normal specimens were blood samples that were compared to liver needle biopsies. Samples were classified according to histological features as non cancerous (10 cases) and pre-cancerous (33 cases, chronic hepatitis and cirrhosis). RESULTS: Our results indicate that there is a tendency of increased chromosomal alteration as lesions become chronic. Samples from patients with antibodies to antibodies for hepatitis C virus show more alterations than hepatitis B positive samples. Steatohepatitis, a disease of unknown etiology, appears to have a high number of microsatellite abnormalities. CONCLUSIONS: Microsatellite APOA2 located on chromosome 1, shows a statistically significant increase in the rate of loss of heterozygosity as liver lesions become more severe, indicating the presence of tumor suppressor genes which may be involved in the development of these lesions. PMID- 10459630 TI - Differential regenerative response and expression of growth factors following hepatectomy of variable extent in rats. AB - AIMS/BACKGROUND: Liver regeneration is a physiological mechanism which leads to restoration of the hepatic parenchyma following hepatectomy or toxic injury. This process is mediated by a wide variety of cytokines and growth factors. The aim of the present study was to evaluate the influence of hepatectomy extent on the levels of intrahepatic mRNAs for cell-cycle markers and growth factors in rats submitted to a 30%, two-third or 80% hepatectomy. METHODS: Cyclins, thymidine kinase and growth factors mRNA levels were quantitatively assessed by RT-PCR at different time points post-hepatectomy (2h, 6h, 12h, days 1, 2, 6). RESULTS: As compared with a two-third hepatectomy, cyclins and thymidine kinase mRNA levels were increased but with a delayed peak at day 2 in the 80% hepatectomy group and showed a progressive increase until day 6 in the 30% hepatectomy group; mRNA levels for HGF or TGFalpha were increased with a delayed peak at 12 h or day 2 in the 80% hepatectomy group, respectively and this delay was more pronounced in the 30% hepatectomy group with a peak at day 1 or day 6. CONCLUSION: A regenerative response occurs whatever the extent of hepatectomy but the course of regeneration and expression of growth factors differs according to the volume of resected liver. A better knowledge of these events could improve the clinical results of hepatic resection for primary or metastatic liver disease. PMID- 10459631 TI - Transforming growth factor-alpha and epidermal growth factor receptor in chronic liver disease and hepatocellular carcinoma. AB - AIMS/BACKGROUND: Transforming growth factor-alpha (TGF-alpha) is a potent mitogen of normal and neoplastic hepatocytes. In addition, TGF-alpha has been reported to play a pivotal role in hepatocarcinogenesis. To evaluate the significance of TGF alpha in chronic liver diseases and hepatocellular carcinoma, we examined serum TGF-alpha, and expression of TGF-alpha, epidermal growth factor receptor (EGFR) and proliferating cell nuclear antigen (PCNA) mRNA in liver tissues. METHODS: Thirty-five patients with chronic hepatitis (CH), 33 with liver cirrhosis (LC), 55 with hepatocellular carcinoma (HCC) and 53 normal controls (C) were enrolled in this study. Serum TGF-alpha levels were measured by an enzyme-linked immunosorbent assay. Expression of TGF-alpha, EGFR, PCNA and beta-actin mRNA in liver tissues were examined by reverse transcription polymerase chain reaction. RESULTS: Serum TGF-alpha levels in C, CH, LC and HCC were 5.6+/-2.1, 33.2+/-8.3, 404.0+/-173.0 and 100.3+/-39.2 pg/ml, respectively. Serum TGF-alpha level in LC was higher than in other diseases (p<0.01, compared to CH, HCC and C, respectively). Serum TGF-alpha levels exhibited a significant positive correlation with total bilirubin, ICGR15 and Pugh score (p<0.01, p<0.01 and p<0.05, respectively), and increased in parallel with severity of disease according to Child classification. Although the ratios of TGF-alpha, EGFR and PCNA mRNA to beta-actin mRNA were not significantly different among the diseases, the TGF-alpha/beta-actin ratio correlated with EGFR/beta-actin and PCNA/beta actin ratios (p<0.001 and p<0.0001, respectively), and EGFR/beta-actin ratio was related to PCNA/beta-actin ratio in all patients, especially with HCC. CONCLUSION: The results of the present study suggest that serum TGF-alpha levels are closely related to severity of liver dysfunction, and that hepatic expression of TGF-alpha and EGFR correlates with proliferation of normal and neoplastic hepatocytes. PMID- 10459632 TI - Effects of dietary iron overload on progression in chemical hepatocarcinogenesis. AB - AIM: The present study was undertaken to investigate possible effects of dietary iron during the progression step in hepatocarcinogenesis. METHODS: Two experiments were performed, in which preneoplastic foci were produced in rat liver using the Solt & Farber protocol, with diethylnitrosamine as initiator and partial hepatectomy + 2-acetylaminofluorene as promoter. Two weeks after promotion, animals were fed 1.25-2.5% dietary carbonyl iron or a control diet until sacrifice. In the first experiment, animals were killed at different time points when they developed an abdominal mass in combination with weight loss. In the second experiment, animals were sacrificed 45 weeks post-promotion. Liver tumours were counted and histologically graded. Tumour levels of ubiquinone-9 and alpha-tocopherol were determined with HPLC, and labelling and apoptotic indices calculated using immunohistochemistry. The number and area of glutathione S transferase 7,7 (GST-7,7)-positive foci were determined. RESULTS: In experiment number 1, survival and tumour differentiation were similar in iron-treated animals and controls. In the second experiment, iron-treated rats had an increased number of GST-7,7-positive foci compared to controls. Number and size of carcinomas were similar between the groups, whereas tumour differentiation was higher in rats exposed to iron. Cell proliferation, apoptosis and concentrations of alpha-tocopherol in tumours were not altered by iron. The ratio of reduced/oxidized ubiquinone-9 was decreased in tumours from iron-treated animals. CONCLUSION: In this model, dietary iron overload resulted in an increased number of preneoplastic foci but did not enhance the progression of these into hepatocellular carcinomas. Iron decreased the ratio of reduced/oxidized ubiquinone-9 in tumours, indicating that neoplastic liver cells utilize intracellular ubiquinones as a defense mechanism against iron-induced oxidative stress. PMID- 10459633 TI - Persistent infection of hepatitis C virus in the elderly: a clinical and quantitative pathological study of autopsy cases. AB - AIMS/BACKGROUND: The entire spectrum of persistent infection by hepatitis C virus (HCV) is still unknown. METHODS: A total of 37 autopsy cases, positive with second generation anti-HCV ELISA, were analyzed. The cases comprised 15 males and 22 females, the average ages being 76.5 in males and 81.1 in females. Twenty-one cases were found to be asymptomatic, while 16 were symptomatic. RESULTS: Based on two pathological parameters of hepatic fibrosis and inflammation, the cases were divided into five groups; group A (non-inflammatory group without significant fibrosis; 11 cases), group B (inflammatory group without significant fibrosis; 9 cases), group C (non-inflammatory group with significant fibrosis; 1 case), group D (inflammatory group with significant fibrosis; 11 cases) and group E (undetermined inflammatory index; 5 cases). All cases in group A and seven cases in group B were asymptomatic. Group A included 6 cases with normal liver. All cases in groups C and D were symptomatic. All examined cases of the inflammatory groups were positive for serum HCV-RNA. CONCLUSION: Anti-HCV-positive elderly autopsy patients include many asymptomatic cases. The two pathological parameters of hepatic fibrosis and inflammation can be used to divide the cases into five groups with each group being well correlated with clinical and virological features. PMID- 10459634 TI - Portal hypertension secondary to arterio-portal fistulae: two unusual cases. AB - A 62-year-old male presented with variceal haemorrhage. Investigation demonstrated a fistula between the left gastric artery and portal vein with a porto-systemic gradient of 35 mm Hg. Variceal bleeding was controlled by a transcatheter embolisation of the fistula, but the patient died of septicaemia three weeks later. The second patient, a 42-year-old male who presented with variceal bleeding was shown to have diffuse arterio-venous fistulae involving the right lobe of the liver with a portosystemic gradient of 25 mm Hg. In this case the variceal bleeding was successfully controlled by insertion of a transjugular intrahepatic portosystemic shunt (TIPS). The pathogenesis of portal hypertension in arterioportal fistulae and the role of interventional radiological techniques in the management of variceal bleeding in these patients is discussed. PMID- 10459635 TI - Clearance of HCV RNA in a chronic hepatitis C virus-infected patient during acute hepatitis B virus superinfection. AB - The routes of hepatitis B virus and hepatitis C virus transmission are quite similar and coexistence of both viruses in one patient is not a rare phenomenon. Until now, the natural course of liver diseases induced by coinfections has not been well documented and the mechanisms of interaction between the two viruses and the human host have not been fully clarified. We report the case of a patient suffering from chronic hepatitis due to hepatitis C virus who developed an acute hepatitis B virus superinfection. Serum hepatitis C virus ribonucleic acid became undetectable by reverse transcriptase/polymerase chain reaction at diagnosis of acute hepatitis B virus infection. At the same time, there was a striking increase in the serum concentrations of the antibodies against C22 and C33c hepatitis C virus antigens. Four months after clinical resolution of the acute hepatitis, hepatitis B surface antigen was undetectable in serum and three months later antibodies against hepatitis B surface antigen appeared. Two years after acute hepatitis B virus infection, the patient has had no relapse of markers for viral replication of hepatitis B virus. Transaminases are within the reference range and hepatitis C virus ribonucleic acid is undetectable in both serum and liver tissue. We hypothesize that acute hepatitis B virus infection stimulated a specific humoral response against hepatitis C virus as well as triggering non specific defense mechanisms which finally eliminated both viruses. PMID- 10459636 TI - Invasive meningococcal disease among university undergraduates: association with universities providing relatively large amounts of catered hall accommodation. AB - The incidence of invasive meningococcal disease (IMD) among UK university students and non-students of similar age was investigated. In addition, we sought to identify structural risk factors associated with high rates of IMD in individual universities. Cases were ascertained via Consultants in Communicable Disease Control (or equivalent officers) between September 1994 and March 1997. Data on individual universities were obtained from university accommodation officers. University students had an increased annual rate of invasive meningococcal disease (13.2/10(5), 95% CI 11.2-15.2) compared with non-students of similar age in the same health districts (5.5/10(5), CI 4.7-6.4) and in those health districts without universities (3.7/10(5), CI 2.9-4.4). This trend was highly significant. Regression analysis demonstrated catered hall accommodation to be the main structural risk factor. Higher rates of disease were observed at universities providing catered hall places for > 10% of their student population (15.3/10(5), CI 11.8-18;8) compared with those providing places for < 10% of students (5.9/10(5), CI 4.1-7.7). The majority of IMD amongst students was caused by serogroup B organisms. University students in the UK are at increased risk of IMD compared with non-students of a similar age. The incidence of IMD tends to be greatest at universities with a high provision of catered hall accommodation. PMID- 10459637 TI - A case-control study of sporadic infection with O157 and non-O157 verocytotoxin producing Escherichia coli. AB - Potential risk factors for sporadic verocytotoxin-producing Escherichia coli (VTEC) infection in Belgium were investigated in a matched case-control study. Thirty-seven cases, 8 infected with O157 VTEC strains (all eaeA-positive), 29 with non-O157 VTEC strains (13 eaeA-positive and 16 eaeA-negative) and 69 matched controls were interviewed. In a conditional logistic regression analysis, consumption of fish appeared to be a risk factor for infection (adjusted odds ratio (OR) 3.25, P = 0.04). Contact with dogs (OR 0.27, P = 0.04) and consumption of shellfish (OR 0.19, P = 0.05) showed a negative association, corresponding to a decrease in risk. These findings might be explained if low level environmental exposure to VTEC induces protective immunity. Eating raw meat, a frequent habit in Belgium, or hamburgers, or eating in a fast-food restaurant was not more frequently reported by cases than controls. The exposures causing sporadic infections with VTEC, in particular non-O157 strains, may be very different from those which led to outbreaks, and may account for more cases overall. PMID- 10459638 TI - Molecular epidemiological investigation of an outbreak of Campylobacter jejuni identifies a dominant clonal line within Scottish serotype HS55 populations. AB - Three molecular typing methods, pulsed-field gel electrophoresis (PFGE), ribotyping, and flagellin (flaA) gene typing, were used to discriminate within a group of 28 Campylobacter jejuni, heat-stable serotype 55 (HS55) isolates derived from cases of campylobacter enteritis occurring throughout Scotland, including 9 isolates associated with an outbreak. PFGE was found to be most discriminatory, identifying 6 distinct profiles, followed by ribotyping (5 profiles), and then flagellin gene typing (4 profiles). The coincidence of all three genotypic markers identified a dominant clonal line within the HS55 group, accounting for each of the outbreak strains, and for 9 of the 19 sporadic strains. A second, closely related, clonal line accounted for a further 5 of the sporadic strains, and also included the HS55 reference strain. Identification and monitoring of such clonal lines should facilitate more effective future epidemiological surveillance of C. jejuni. PMID- 10459639 TI - An outbreak of Salmonella typhimurium DT170 associated with kebab meat and yogurt relish. AB - During July 1995, an outbreak of Salmonella typhimurium definitive type (DT) 170, an unusual strain, occurred in South Wales. A case-control study found that illness was associated with eating kebabs (odds ratio undefined, P = 0.002), doner kebabs (odds ratio 7.9, 95 % confidence interval 1.5-20.5, P = 0.02) and kebabs with yoghurt based relish (odds ratio undefined, P = 0.009) but not with eating kebabs with mayonnaise-based relish (odds ratio 2.4, 95 % confidence interval 0.4-13.9, P = 0.53). Environmental investigations discovered a complex web of producers and wholesale suppliers. Kebab meat and yoghurt had been supplied to the two main implicated outlets by a single wholesaler. Samples of raw minced lamb and several environmental swabs taken at the wholesaler were positive for S. typhimurium DT170. Blood-stained, unsealed yoghurt pots were observed to be stored under a rack of raw lamb. Investigators of food poisoning outbreaks linked to takeaway food should consider cross-contaminated relishes and dressings as well as undercooked meat as potential vehicles of infection. PMID- 10459640 TI - Outbreaks of salmonellosis associated with eating uncooked tomatoes: implications for public health. The Investigation Team. AB - Laboratory-based surveillance of salmonella isolates serotyped at four state health departments (Illinois, Michigan, Minnesota and Wisconsin) led to the identification of multistate outbreaks of salmonella infections during 1990 (176 cases of S. javiana) and 1993 (100 cases of S. montevideo). Community-based case control studies and product traceback implicated consumption of tomatoes from a single South Carolina tomato packer (Packer A) MOR 16.0; 95% CI2.1, 120.6; P < 0.0001 in 1990 and again in 1993 (MOR 5.7; 95 % CI 1.5, 21.9; P = 0.01) as the likely vehicle. Contamination likely occurred at the packing shed, where field grown tomatoes were dumped into a common water bath. These outbreaks represent part of a growing trend of large geographically dispersed outbreaks caused by sporadic or low-level contamination of widely distributed food items. Controlling contamination of agricultural commodities that are also ready-to-eat foods, particularly fruits and vegetables, presents a major challenge to industry, regulators and public health officials. PMID- 10459641 TI - Virulence of Salmonella enterica serotype Enteritidis aflagellate and afimbriate mutants in a day-old chick model. AB - Certain fimbriae and the flagellae of Salmonella enterica serovar Typhimurium have been shown to contribute to attachment and invasion of gut epithelium in the murine typhoid infection model and to contribute to pathogenesis in the chick. However, little is known of the role these organelles play in Enteritidis poultry infections and, to study this, day-old chicks were dosed orally in separate experiments with defined multiply afimbriate and/or aflagellate mutant strains of Enteritidis. The colonization and invasion characteristics of each mutant were compared with those of the isogenic wild type strain by the determination of the number of bacteria recovered from livers and spleens at known time points post infection. Compared with wild type Enteritidis, a mutant unable to express flagella but retaining the genetic potential to express fimbriae was recovered post mortem from livers and spleens in significantly reduced numbers compared to the isogenic wild-type at all time points post infection (P < 0.001). Conversely, a flagellate but multiply afimbriate mutant (defective for the elaboration of five different fimbrial types) and a flagellate but non-motile 'paralysed' mutant were recovered from livers and spleens in similar numbers to the wild-type. The data suggested that Enteritidis flagella, but not fimbriae, played an important role in pathogenesis in the chick model and that the flagellar apparatus itself and not motility per se contributed significantly to this role. PMID- 10459642 TI - Health risks associated with unpasteurized goats' and ewes' milk on retail sale in England and Wales. A PHLS Dairy Products Working Group Study. AB - A pilot study to determine the microbiological quality of unpasteurized milk from goats and ewes sampled from farm shops, health food shops, and other retail premises found that 47%, (47/100) of goats' and 50% (13/26) of ewes' milk samples failed the standards prescribed by the Dairy Products (Hygiene) Regulations 1995. In addition, Staphylococcus aureus, haemolytic streptococci or enterococci, were present in excess of 10(2) c.f.u./ml in 9 (7 %) 2 (2 %) and 19 (15%) samples, respectively. Salmonella, campylobacter, verocytotoxin-producing Escherichia coli O157:H7 and Listeria monocytogenes were not detected in the samples. At the time of purchase approximately half of the milk samples (58 %) were frozen, the rest were liquid. Farm outlets sold predominantly liquid milk, other retail premises sold a frozen product. The microbiological quality of goats' and ewes' milk, whether frozen or liquid, was not significantly different. Milk sold from farm shops was of lower quality than that from health food shops and other retail premises. In this pilot study most producers (92 %) supplied, and most retailers (76 %) sold unpasteurized goats' and ewes' milk that contained unacceptable levels of indicator organisms. The study was carried out during the winter when goats' milk production is reduced. The results indicate the need for a full representative study of unpasteurized goats' and ewes' milk on retail sale throughout the year. PMID- 10459643 TI - Surveying vendors of street-vended food: a new methodology applied in two Guatemalan cities. AB - Lack of reliable data about street vendors, who are difficult to survey, has hampered efforts to improve the safety of street-vended food. A two-phase method for sampling vendors, surveying first in areas of concentrated vending activity identified by local authorities and second in randomly selected areas, was developed and implemented in two Guatemalan cities where street-vended food had been implicated in cholera transmission. In a 4-day survey in Escuintla, 59 vendors (42 from phase 1, 17 from phase 2) were interviewed. They demonstrated good knowledge of food safety and cholera but unsafe practices, implying that more effective, practical training was needed. In a 6-day survey in Guatemala City, 78 vendors (77 from phase 1, 1 from phase 2) were interviewed. Sixty-eight (87 %) vendors stored water, usually in wide-mouthed vessels prone to contamination; this led to a field test of a new system for safe water storage. Useful information for public health planning and intervention can be gathered rapidly with this new method for surveying street vendors. PMID- 10459644 TI - DNA restriction patterns produced by pulsed-field gel electrophoresis in Moraxella catarrhalis isolated from different geographical areas. AB - Pulsed field gel electrophoresis (PFGE) of the genomic DNA of Moraxella catarrhalis was done in 172 strains isolated from sputum of patients with respiratory infections in Nagasaki (130 strains), Europe (14 strains), Thailand (6 strains), Uganda (3 strains), Bangladesh (5 strains) and Kuwait (14 strains). Restriction endonuclease with SmaI generated 4-16 DNA fragments ranging from 1000 kb to 24.25 kb and was classified into 31 major groups. It was found that there were wide variations of DNA restriction patterns of strains isolated from the same and different geographical areas. DNA restriction patterns of strains isolated in Nagasaki during the last 12 years showed dynamic changes of the predominant strains in each time period. We conclude from this study that PFGE is a suitable method to document interstrain variation in M. catarrhalis. PMID- 10459645 TI - Leptospiral infection among primitive tribes of Andaman and Nicobar Islands. AB - The Andaman islands were known to be endemic for leptospirosis during the early part of the century. Later, for about six decades no information about the status of the disease in these islands was available. In the late 1980s leptospirosis reappeared among the settler population and several outbreaks have been reported with high case fatality rates. Besides settlers, these islands are the home of six primitive tribes of which two are still hostile. These tribes have ample exposure to environment conducive for transmission of leptospirosis. Since no information about the level of endemicity of the disease among the tribes is available, a seroprevalence study was carried out among all the accessible tribes of the islands. A total of 1557 serum samples from four of the tribes were collected and examined for presence of antileptospiral antibodies using Microscopic Agglutination Test (MAT) employing 10 serogroups as antigens. An overall seropositivity rate of 191% was observed with the highest rate of 53.5% among the Shompens. The seropositivity rates in the other tribes were 16.4% among Nicobarese, 222% among the Onges and 14.8% among the Great Andamanese. All of the tribes except the Onges showed a similar pattern of change in the seroprevalence rates with age. The prevalence rates were rising from low values among children to reach a peak in those aged 2140 years and then declined. Among Onges the seroprevalence rates continued to rise beyond 40 years. In all the tribes, seroprevalence rates were found to be significantly higher among the males. The commonest serogroups encountered were Australis followed by Grippotyphosa, Icterohaemorrhagiae, Pomona and Canicola. PMID- 10459646 TI - Antibody response to OspC-I synthetic peptide derived from outer surface protein C of Borrelia burgdorferi in sera from Japanese forestry workers. AB - The prevalence of antibodies against Lyme disease spirochaetes in serum samples from 80 forestry workers at high occupational risk of Lyme disease was surveyed by enzyme-linked immunosorbent assay (ELISA) with the OspC-I synthetic peptide. The peptide is part of the outer surface protein C (OspC) amino acid sequence located in the region conserved among Borrelia burgdorferi sensu stricto or sensu lato. Positivity for antibodies against OspC-I was observed in 25 (313%) of the forestry workers. Of these positive cases, 12 (15.0%) and 19 (23.8%) were positive for immunoglobulin M (IgM) and IgG antibody, respectively. Among 62 workers who were negative for IgG antibody against B. garinii or B. japonica in our previous study, 9 (14.5%) and 4 (6.5%) were positive for IgM and IgG antibody, respectively, in OspC-I ELISA. These results demonstrate for the first time that Lyme disease in forestry workers can be revealed using OspC-I ELISA. We conclude that forestry workers who show positive results for antibodies against OspC-I have very likely been exposed to Lyme disease spirochaetes, and that those who show positivity for IgM antibody against OspC-I may be in the early stage of Lyme disease. PMID- 10459647 TI - Molecular epidemiology of Mycobacterium avium complex isolated from patients with and without AIDS in Brazil and England. AB - Mycobacterium avium complex (MAC) is ubiquitous throughout the world. It is an opportunistic pathogen in AIDS patients but the number of cases in HIV negative patients is also increasing. The aim of this study was to determine whether patients were being infected with different MAC strains or whether one strain was dominant. DNA obtained from isolates in Brazil and England were compared using pulsed field gel electrophoresis (PFGE). Strains from 22 Brazilian patients clustered into 7 groups but 68/90 patients had a unique strain. In all patients, Brazilian and English, the same strain was isolated repeatedly over time, some over several years. This study shows that it is most likely that Man is infected from the environment and that one strain can survive without change for many years both in the environment and in Man. PMID- 10459648 TI - Measles in Vietnamese refugee children in Hong Kong. AB - From September 1991-January 1992, there was a measles epidemic in an established refugee camp for 7000 Vietnamese 'Boat People' living in Hong Kong. This 16 week outbreak occurred against a backdrop of poor uptake of measles vaccination and overcrowded living conditions. Two hundred and sixty-two children were affected (155 boys, 107 girls); 235 children (89.7 %) were < 2 years old, age range 5-39 months. Children age 6-11 months had the highest crude attack rate (AR) of 54.3%. The highest age specific crude AR was 83.8% in children aged 14 months. Measles complications affected 234 (89.3%) children. Sixty-six children (25.2%) were admitted to hospital. There were two deaths from pneumonia, giving a case fatality rate of 0.76%. Measles control in refugee camps continues to be a public health challenge. PMID- 10459649 TI - A major outbreak of hantavirus infection in Belgium in 1995 and 1996. AB - Haemorrhagic fever with renal syndrome (HFRS) is a human disease characterized by flu-like symptoms, renal dysfunction, and in severe cases, haemorrhagic manifestations. The causative agents of HFRS are Hantaan (HTN), Seoul (SEO), Puumala (PUU) and Dobrava (DOB) hantaviruses. Hantavirus infections are of increasing importance in Europe. Outbreaks occur in Belgium with a 3- to 4-year interval with an increasing number of cases. We describe the largest outbreak so far in Belgium with 217 serologically and clinically confirmed cases in the period between October 1995 and December 1996. We demonstrated that the use of viral antigen derived from a local PUU-strain was able to detect significantly more sera positive for IgM in an immunofluorescence assay. Furthermore, although in some cases SEO, HTN and DOB antibody-reactivities were detected by ELISA, only PUU infections could be confirmed by neutralization test. The presence of an unknown hantavirus serotype circulating in Belgium should be considered. PMID- 10459650 TI - Cowpox: reservoir hosts and geographic range. AB - It is generally accepted that the reservoir hosts of cowpox virus are wild rodents, although direct evidence for this is lacking for much of the virus's geographic range. Here, through a combination of serology and PCR, we demonstrate conclusively that the main hosts in Great Britain are bank voles, wood mice and short-tailed field voles. However, we also suggest that wood mice may not be able to maintain infection alone, explaining the absence of cowpox from Ireland where voles are generally not found. Infection in wild rodents varies seasonally, and this variation probably underlies the marked seasonal incidence of infection in accidental hosts such as humans and domestic cats. PMID- 10459651 TI - Risk factors for Plasmodium vivax infection in the Lacandon forest, southern Mexico. AB - A study was conducted to characterize the risk of Plasmodium vivax infection in the Lacandon forest, southern Mexico. Blood samples and questionnaire data were collected in 1992. Malaria cases (n = 137) were identified by the presence of symptoms and a positive thick blood smear. The control group included individuals with negative antibody titres and no history of malaria (n = 4994). From 7628 individuals studied, 1006 had anti-P. vivax antibodies. Seroprevalence increased with age. Risk factors associated with infection included: place of birth outside the village of residence (odds ratio, OR 11.67; 95% CI 5.21-26.11); no use of medical services (OR 4.69, 95% CI 3.01-7.29), never using bed-nets (OR 3.98, 95 % CI 1.23-12.86) and poor knowledge of malaria transmission, prevention and treatment (OR 2.30, 95 % CI 1.30-4.07). Health education represents the best recommendation for controlling the disease in the area. PMID- 10459652 TI - The epidemiology of head lice and scabies in the UK. AB - Anecdotal evidence suggests that the prevalence of both scabies and head lice is increasing and also that both conditions are becoming refractory to pesticide treatment. Using information obtained from the Office of National Statistics, Royal College of General Practitioners Weekly Returns Service, Department of Health, local surveys of school children from Bristol and drug sales of insecticides, we have confirmed that there has been a rise in the prevalence of both conditions. We have shown that scabies is significantly more prevalent in urbanized areas (P < 0.00001), north of the country (P < 0.000001), in children and women (P < 0.000001) and commoner in the winter compared to the summer. Scabies was also shown to have a cyclical rise in incidence roughly every 20 years. Head lice were shown to be significantly more prevalent in children and mothers (P < 0.000001) though both conditions were seen in all age groups. Head lice were also less common during the summer. Host behaviour patterns, asymptomatic carriage, drug resistance and tourism from countries or districts with a higher incidence may be important factors in the currently high prevalence of both scabies and head lice. PMID- 10459653 TI - Genetic subtyping of Escherichia coli O157 isolates from 41 Pacific Northwest USA cattle farms. AB - Escherichia coli O157 (n = 376) from 41 cattle farms were subtyped using pulsed field gel electrophoresis of endonuclease cleaved chromosomal DNA. Cleavage with XbaI resulted in 81 subtypes. Fifty-one isolates from subtypes found in more than one herd, or in herds on multiple sample collection dates were compared using the endonuclease NotI, resulting in 23 additional subtypes. Up to 11 XbaI subtypes were found per farm with up to 7 subtypes/farm identified from a single date. Indistinguishable subtypes (both XbaI and NotI) were found to persist on 4 farms for 6-24 months. Five subtypes were found on more than one farm separated by up to 640 km. Dairy farms where cattle had moved onto the farm had a similar number of subtypes as farms with no movement of cattle, and feedlots had more subtypes than dairy farms. These data indicate that there is a mechanism for multiple herd exposure to specific subtypes, there are multiple sources of exposure for cattle on farms, and on-farm reservoirs other than cattle may exist. PMID- 10459654 TI - Sampling considerations for herd-level measurement of faecal Escherichia coli antimicrobial resistance in finisher pigs. AB - The objective of this study was to determine the most efficient means of sampling faeces of finisher pigs for accurate and precise farm-level estimates of antimicrobial resistance among faecal Escherichia coli. Resistance to tetracycline and gentamicin of 8250 isolates of E. coli from 55 finisher pigs on one farm was measured with a hydrophobic grid membrane filter method. The between pig, within-pen component of variance in resistance was large (97.5%), while between-pen, within-room and between-room components were small (2.5% and 0%, respectively). Using these resistance data, the abilities of two sampling strategies to estimate prevalence were modelled with a Monte Carlo 'bootstrap' procedure. Compositing faecal samples from several pigs before testing produced unbiased and precise estimates of prevalence and is simpler technically than individual animal testing. PMID- 10459655 TI - Discrimination between endemic and feedborne Salmonella Infantis infection in cattle by molecular typing. AB - Salmonella enterica serovar Infantis is endemic in Finnish cattle. Feed contaminated with S. Infantis was distributed to cattle farms in May 1995. Following increased sampling, S. Infantis was detected on 242 farms in 1995. Molecular typing was used to differentiate the farms that were infected by the feed-related Infantis from those infected by other endemic strains. Twenty-three isolates from feed in 1995 and 413 from cattle (72 from 19924, 324 from 1995, 17 from 1996-7) were analysed. The feed-related Infantis was clonally related to the endemic infection by the ribotype, IS200-type and XbaI-profile. The feed isolates had a distinctive plasmid that appeared in pulsed-field gel electrophoresis as a 60 kb band when cleaved with XbaI or linearized by S1-nuclease. This plasmid appeared in cattle only since the outbreak and seemed stable on the follow-up farms. In addition to contact farms, the feedborne strain was found on 19% of the farms infected with S. Infantis in 1995 but not having bought suspected feedstuffs, possibly as secondary infections. PMID- 10459656 TI - A comparison of wildlife control and cattle vaccination as methods for the control of bovine tuberculosis. AB - The Australian brushtail possum is the major source of infection for new cases of bovine tuberculosis in cattle in New Zealand. Using hypothetical values for the cost of putative cattle and possum Tb vaccines, the relative efforts required to eradicate Tb in cattle using possum culling, possum vaccination or cattle vaccination are compared. For realistic assumed costs for 1080 poison bait, possum culling is found to be a cost-effective strategy compared to cattle vaccination if the required control area is below 13 ha per cattle herd, while possum vaccination is cost-effective for control areas of less than 3 ha per herd. Examination of other considerations such as the possible roles of possum migration and heterogeneities in possum population density suggest that each control strategy may be superior under different field conditions. Finally, the roles of the possum in New Zealand, and the Eurasian badger in Great Britain and Ireland in the transmission of bovine tuberculosis to cattle are compared. PMID- 10459658 TI - Genetic analysis of type O viruses responsible for epidemics of foot-and-mouth disease in North Africa. AB - The nucleotide sequences of the 3' end of the capsid-coding region were determined for 30 serotype O foot-and-mouth disease (FMD) viruses isolated between 1987 and 1994 from outbreaks in North Africa and the Middle East. These sequences were compared with the previously published sequences of 9 field virus isolates from the Middle East and 5 vaccine virus strains, 3 of which originated from the Middle East (O1/Turkey/Manisa/69, O1/Sharquia/Egypt/72 and O1/Israel/2/85) and 2 from Europe (O1/Lausanne/Switzerland/65 and O2/Brescia/Italy/47). Cluster analysis of these sequences using the unweighted pair group mean average (UPGMA) method showed: (i) that the FMD viruses isolated from North Africa and the Middle East were very different from the classical European vaccine strains; (ii) that all the viruses isolated during the 1989-92 North African epidemic formed a cluster differing by no more than 6% from each other; (iii) a virus isolated in Libya in 1988 was unrelated to the aforementioned epidemic; and (iv) viruses from a second, less extensive epidemic, occurring in 1994, fell into yet another cluster. PMID- 10459657 TI - PCR-RFLP of outer membrane proteins gene of Dichelobacter nodosus: a new tool in the epidemiology of footrot. AB - Currently only phenotypic epidemiological markers, serogrouping and virulence testing of Dichelobacter nodosus, are available for investigating footrot outbreaks in small ruminants. These methods have limitations in tracing the source of infection. In this study, a genotypic marker, PCR-RFLP of outer membrane protein gene, was used to characterize D. nodosus. The technique was evaluated in a controlled experiment involving two strains of bacteria. PCR-RFLP was found to be highly specific in differentiating isolates obtained from recipient animals infected with different strains. Subsequently, this technique was used to characterize isolates obtained from field cases of footrot in Nepal. A total of 11 patterns was recognized among 66 Nepalese D. nodosus isolates representing four different serogroups. PCR-RFLP also discriminated isolates with similar phenotypic characteristics. However, all isolates which, phenotypically, were virulent were represented by only two patterns irrespective of their serogroups. It is suggested that PCR-RFLP described here could be a useful epidemiological marker in the study of footrot. PMID- 10459659 TI - Predicting the level of herd infection for outbreaks of foot-and-mouth disease in vaccinated herds. AB - Foot-and-mouth disease (FMD) is a highly contagious virus infection of sheep, goats, cattle, pigs and other, non-domesticated species of artiodactyls, and causes both clinical and subclinical infection according to the natural or acquired immunity of the host. Within vaccinated dairy herds FMD may appear as an acute, mild or subclinical infection, dependent upon the immune status of the herd, the level of challenge and the efficacy of the vaccine used. In the large dairy herds of Saudi Arabia, sub-clinical FMD was on a number of occasions, found to have spread amongst the cattle before signs of disease were seen. Such undetected transmission resulted in a large incidence on the first day of diagnosis and curtailed the impact of post-outbreak vaccination (PoV). First day incidence (FDI) for these herds was found to correlate with the final cumulative incidence of clinical disease. Since FDI is available at the start of an outbreak it can be used as a predictive tool for the eventual outcome of an FMD outbreak. During the past 11 years 47 % of dairy herds examined in Saudi Arabia have experienced FMD initially as sub-clinical disease. For the remaining 53 %, waning vaccinal protection did not suppress clinical disease in the initially infected animals, and these showed severe rather than mild signs. Hence, in such herds there was a very low initial level of subclinical infection, so PoV was more effective, and the timing of PoV was found to give a good correlation with cumulative herd incidence: an early PoV resulted in low prevalence of clinically infected animals whilst late PoV permitted high prevalence. PoV timing can thereby be used in tandem with FDI as a predictive tool for future outbreaks, estimating the final cumulative incidence (or prevalence) of clinical FMD cases. PMID- 10459660 TI - Pigs with highly prevalent antibodies to human coronavirus and swine haemagglutinating encephalomyelitis virus in the Tohoku District of Japan. AB - From 1985 to 1988, a total of 2496 swine sera from 60 farms in the Tohoku District of the Honshu Island of Japan were examined for antibodies to swine haemagglutinating encephalomyelitis virus (HEV), human coronavirus (HCV) and bovine coronavirus (BCV) by haemagglutination-inhibition (HI) test. Antibodies to HEV 67N strain and HCV OC43 strain were highly prevalent with positivity rates of 82.1 and 91.4%, respectively, while seropositivity rate to BCV Kakegawa strain was 44.2%. No clinical signs of HEV infection were noticed in any farms including farms with relatively high seropositivity. The results suggested that HCV or antigenitically related virus(es) as well as HEV might be perpetuated in swine in the Tohoku District. PMID- 10459661 TI - Classical swine fever in Sardinia: epidemiology of recent outbreaks. AB - A variable region of the gene encoding the major glycoprotein (E2) of Classical Swine Fever Virus (CSFV) was sequenced from 12 Sardinian isolates which had been obtained from three geographically distinct regions of the Island. Phylogenetic analysis of these viruses and others characterized in previous studies [1, 2] indicated that (a) the Sardinian viruses were all members of the common European subgroup 2.3 and were clearly distinct from live vaccines recently used in this area; (b) they could be resolved into four distinct groups in accordance with the region or date of isolation; (c) in at least two regions wild boar/domestic swine contact was implicated in virus spread; (d) the oldest isolate (1983) and some of the recent isolates were possibly introduced from mainland Italy. In addition, this study has wider implications for the interpretation of CSFV variation. We have been able to demonstrate that small variations within this region of the virus genome (possibly less than 2.7% or five nucleotide substitutions) can be used to separate isolates into groups that precisely fit their geographical distribution. This finding is especially important for deducing the epidemiological relationships between multiple outbreaks caused by similar viruses that occur in close proximity. PMID- 10459662 TI - Effects of prolactin on sleep in cyclic rats. AB - We previously demonstrated that pregnancy-associated sleep enhancement is correlated with the daily surges of prolactin (PRL). However, in spite of a surge of PRL in the proestrous night, a reduction of nocturnal sleep occurs in phase with proestrus. Therefore, to clarify the physiological role of PRL in sleep regulation during the estrous cycle, time-course changes in sleep were analyzed in bromocriptine (CB-154)-treated and/or PRL-supplemented female rats. Sleep patterns characteristic of proestrus-to-estrus were not affected by the CB-154 treatment. In contrast, nocturnal rapid eye movement sleep (REMS) was significantly increased after the PRL supplementation. The CB-154 treatment diminished the REMS-enhancing effect of PRL. Thus, the results suggest that the endogenous PRL is not crucial for the regulation of sleep during the estrous cycle, while exogenous PRL can enhance REMS. PMID- 10459663 TI - Colony-stimulating factors in rapid eye movement sleep and non-rapid eye movement sleep regulation. AB - Although several cytokines are known to be somnogenic, no study has been conducted to examine whether colony-stimulating factors (CSF) affect sleep. Therefore, we studied the effects of granulocyte-macrophage CSF (GM-CSF) and macrophage CSF (M-CSF) on sleep in rats and their possible mechanism of action. At the dose of 10 pmol, GM-CSF or M-CSF significantly increased both non-rapid eye movement and rapid eye movement (REM) sleep or REM sleep only when infused intracerebroventricularly during the dark period. When injected locally in the hypothalamus, GM-CSF and M-CSF increased nitric oxide (NO) production. Thus, NOergic neural signals in the hypothalamus may take part in the somnogenic action of CSF. PMID- 10459664 TI - Acetylcholine and glutamate release during sleep-wakefulness in the pedunculopontine tegmental nucleus and norepinephrine changes regulated by nitric oxide. AB - Cholinergic neurons in the pons appear to play a major role in generating rapid eye movement (REM) sleep. In the present study, acetylcholine and glutamate release in the pedunculopontine tegmental nucleus (PPT) during the sleep-waking cycle were investigated by in vivo microdialysis. Acetylcholine release during slow wave sleep (SWS) was significantly lower (P<0.05) than during REM sleep and wakefulness. On the other hand, glutamate release during wakefulness was higher (P<0.05) than during REM sleep and SWS. Furthermore, the application of N-methyl D-aspartate (1 mM) induced a significant increase of nitric oxides (NOx) for 20 min (P<0.05) and a decrease of norepinephrine for the first 15 min (P=0.01), indicating NOx regulation on norepinephrine release in PPT. PMID- 10459665 TI - The effect of 2-h sleep reduction by a delayed bedtime on daytime sleepiness in children. AB - The purpose of the present study was to investigate the effect of acute 2h sleep reduction by a delayed bedtime on daytime sleepiness. Three children were evaluated over three consecutive days: two control days and a 2h sleep reduction day. A sleep latency test was conducted five times each day at 2h intervals from 10.00 h. The results showed that the effects of sleep loss generally increased daytime sleepiness, in particular, at 10.00 h, and suggested that the intensity of morning sleepiness may become a useful measure which reflects sleep loss. PMID- 10459666 TI - Skin blood flow responses during sleep by laser Doppler flowmetry. AB - Skin surface blood flow at the fingertips was measured in a 21-year-old woman during sleep using laser Doppler flowmetry. Skin blood flow responses (SBFR) are transient reductions in skin blood flow of about 25s in duration. In our subject, SBFR showed a delay time of 8-9s from the onset of skin potential responses (SPR) and appeared more consistently than SPR. The frequency of SBFR decreased during non-rapid eye movement sleep, especially when the sleep stage became deep. These findings indicate the validity of SBFR as a discrete index of autonomic function during sleep. PMID- 10459667 TI - Sleep stage and skin temperature regulation during night-sleep in winter. AB - To clarify the thermally comfortable environment for satisfied sleep, we investigated the relationship between skin temperatures and sleep stages during night-sleep in winter, summer and autumn. Four healthy young females served as the subjects. The experiments were performed in a climate chamber under three conditions: at 13 degrees C-RH70% with futon and blanket in winter, at 20-18 degrees C-RH60% with futon and towelket in autumn and at 29 degrees C-RH70% with towelket in summer. The subjects' skin temperature on the sole was lower until about 4 hours after lights off in winter than in autumn and summer, and the percentage of st.2 was lower and st.4 was higher in winter than in autumn and summer, although the bed climate both in winter and autumn was much the same. PMID- 10459668 TI - Sleep patterns during 20-m nitrox saturation dives. AB - The sleep patterns were examined through three simulated 20-m nitrox saturation dives. The standard polysomnography of 12 divers was recorded respectively for a total of 204 nights, as were patterns of change or consistency in sleep variables. For the 11 divers in their 20s and 30s there was nothing unusual about their sleep variables. However, a reduction of total sleep time in accordance with the lengthening of sleep latency was observed. This was recognized from the latter part of the bottom period to the postdive period. This tendency was notable for the diver in his 50s. These findings suggest that the decompression environment and the psychological stress of the long-term closed environment of a hyperbaric chamber have effects on divers' sleep. PMID- 10459669 TI - A study on polysomnographic observations and subjective experiences under sensory deprivation. AB - We investigated the relationship between subjective experiences and polysomnographic observations under sensory deprivation (SD). The results can be divided into two types: (i) visual images were perceived frequently, accompanied by alpha waves or theta waves in electroencephalogram and frequent rapid eye movements (REM); (ii) the subjects fell in deep sleep without perception of visual images. The perceptions of visual images with alpha waves or theta waves and REM are supposed to be different from those in usual wakefulness and sleep. These results suggest that the SD could disclose different aspects of consciousness from the conventionally categorized states such as wakefulness and sleep. PMID- 10459670 TI - Changes in sleep-wake cycle during the period from late pregnancy to puerperium identified through the wrist actigraph and sleep logs. AB - The purpose of this study was to understand the sleep-wake cycle during the period from late pregnancy to about 3 months of postpartum by evaluating the number of actigraphic activities in four women (one multipara and three primi gravidae), and to compare the results with the findings from sleep logs. An irregularity of the sleep-wake cycle with increased number of wakings at night was notable during about 1 month after delivery, compared to the late pregnancy period, and subsequently this number tended to decrease. These results were indicative of the association between the lactation cycle to neonates and the sleep-wake cycle. PMID- 10459671 TI - Analyses of mothers' sleep logs in postpartum periods. AB - In order to investigate mothers' sleep-wake patterns from five to 12 weeks in the postpartum period, we analyzed continuous sleep logs. Data for 341 days from seven mothers were analyzed for each postpartum week. There were significant differences in total sleep time and wake time at night. Wake time at night gradually decreased from the fifth to the twelfth week. These results from continuous sleep logs support the results of our previous polysomnographic findings based on intermittent measurements. A transition from interrupted sleep to noninterrupted sleep was observed from the ninth to twelfth week of postpartum. The mothers' sleep-wake patterns are discussed focusing on their relationships to the infants' sleep-wake patterns and feeding rhythm. PMID- 10459672 TI - Classification of the sleeping pattern of normal adults. AB - A questionnaire concerning their sleep-related lifestyle and mental health was mailed to people aged in their twenties to fifties living in the Fukushima (North East), Tokyo (Central, Metropolitan) and Okayama (West) areas of Japan. We classified the sleeping pattern of 3642 people (1702 men, 1940 women; mean age 41.3+/-10.67) by multivariational analyses (factor analysis and cluster analysis). They were classified into six groups and defined as 'poor sleeper group' (30.0%), 'good sleeper group' (28.3%), 'long sleeper group' (8.2%), 'short sleeper group' (18.5%), 'irregular sleeper group' (11.3%), and 'Delayed Sleep Phase Syndrome group' (3.7%) based on their characteristics of the factor pattern. PMID- 10459673 TI - A case report on EEG nocturnal sleep in visually impaired persons aged in their 30s and 50s. AB - To determine the EEG sleep structure in visually impaired persons, EEG sleep recordings were made over two or three consecutive nights on five subjects in their 30s and 50s in the laboratory or in the Welfare Center for the Blind. Sleep variables were compared to the normative data of sighted persons of comparable ages. The results indicated that the percentages of slow wave sleep in four of the five blind individuals were much less than the values of the normative data, which is in agreement with Krieger and Glick's results (1971). PMID- 10459674 TI - Laterality of delta waves during all-night sleep. AB - In order to clarify the functional asymmetry of the brain function during sleep, period-amplitude analysis of delta electroencephalogram activity was performed on polysomnograms in 12 right-handed healthy males. Electroencephalograms were recorded from disc electrodes placed at C3, C4, O1 and O2 (10-20 electrode system), using A1 +A2 for reference. Although there were no significant differences in delta counts between O1 and O2, delta counts of C3 were significantly larger than those of C4. These results suggest that there exists distinct laterality in the number of delta waves in the central region, reflecting the functional asymmetry of the brain during sleep. PMID- 10459675 TI - Gender difference of slow wave sleep in middle aged and elderly subjects. AB - Sleep EEG of eight healthy males and eight females aged 54-72 years were recorded at their homes. The electroencephalograms were visually scored and analyzed by spectral analysis using the FFT method. There were no significant differences in sleep parameters except for a higher percentage of stage 3+4 in females. The spectral power of the delta band EEG was classified into two frequencies: 0.5-2 Hz and 2-4 Hz. The total amount of the delta band spectral power through the night was significantly larger in females. Periodic fluctuation of delta band power was observed in females along with non-rapid eye movement-rapid eye movement cycles. PMID- 10459676 TI - Topographic mapping of EEG spectral power and coherence in delta activity during the transition from wakefulness to sleep. AB - The present study examined the topographic characteristics in delta band activity during the transition from wakefulness to sleep, using topographic mapping of electroencephalogram (EEG) power and coherence corresponding to nine EEG stages. The dominant topographic components of delta band activity increased clearly from the vertex sharp-wave stage EEG stage 6) in the anterior-central area. Principal component maps revealed the scalp distribution of EEG. Delta activity in the sleep onset period were composed of two covariant components. One is a diffuse component widely distributed on the scalp and the other is a local component at the temporo-occipital area. PMID- 10459677 TI - Non-linear analysis of the sleep EEG. AB - A sleep electroencephalogram was analyzed by non-linear analysis. The polysomnography of a healthy male subject was analyzed and the correlation dimensions calculated. The correlation dimensions decreased from the 'awake' stage to sleep stages 1-3 and increased during rapid eye movement (REM) sleep. These results were seen during each sleep cycle. In each sleep cycle, the correlation dimensions decrease for slow wave sleep, and increase for REM sleep. PMID- 10459678 TI - Changes of amplitude and topographical characteristics of event-related potentials during the hypnagogic period. AB - In the previous study, during the vertex sharp wave period (hypnagogic EEG stage 4), negative components (N300, N550) were dominant at Fz and Cz in contrast to the positive component (P400) being prominent at the other areas, Pz, Oz, T5 and T6. There is no agreement regarding P400 properties during the hypnagogic period. In this study, using topographic mapping, we found that two negative components (N300, N550) and P400 independently increased their amplitude at the different areas of the scalp as arousal level lowered. The anterior negative components may reflect the information processing related to the K-complex. The P400 may reflect other activities different from the K-complex mechanism or P300 attention mechanisms. PMID- 10459679 TI - Functional changes of the brainstem triggering vertex sharp wave with spindle. AB - A vertex sharp wave followed by spindle (VS-spindle) is one of the hallmarks of human stage 2 sleep. We recorded sleep electroencephalograms (EEG) simultaneously with brainstem auditory evoked potentials (BAEP) from nine healthy male subjects. To investigate the generating mechanism of the VS-spindle, sequentially changing BAEP were analyzed around the VS-spindle. The results revealed the preceding changes of wave-V amplitude to the onset of the VS-spindle. When the generator site of wave-V in the brainstem was considered, the results suggest that the functional changes in the dorsal area of the midbrain-pontine junction participate in the organization of the VS-spindle. PMID- 10459680 TI - Night-day-night sleep-wakefulness monitoring by ambulatory integrated circuit memories. AB - A medium-sized portable digital recorder with fully integrated circuit (IC) memories for sleep monitoring has been developed. It has five amplifiers for EEG, EMG, EOG, ECG, and a signal of body acceleration or respiration sound, four event markers, an 8 ch A/D converter, a digital signal processor (DSP), 192 Mbytes IC flash memories, and batteries. The whole system weighs 1200 g including batteries and is put into a small bag worn on the subject's waist or carried in their hand. The sampling rate for each input channel is programmable through the DSP. This apparatus is valuable for continuously monitoring the states of sleep-wakefulness over 24 h, making a night-day-night recording possible in a hospital, home, or car. PMID- 10459682 TI - Linear automatic detection of eye movements during the transition between wake and sleep. AB - Slow eye movement (SEM) are a sensitive indicator of lowered consciousness or drowsiness in man. A new computerized method for detection of SEM was introduced. A linear regression analysis was applied in each moving window for approximation to the tangent line on the electro-oculogram curve. The results revealed that SEM were more frequent and their duration was shorter at stage wake than at sleep stages 1 and 2. The method was practically suitable for objective detection and measurement of SEM. PMID- 10459681 TI - Development of the polysomnographic database on CD-ROM. AB - We have developed a polysomnographic database on CD-ROM. The data were obtained from 16 subjects with sleep apnea syndrome. The physiological signals include electroencephalogram, electromyogram, electrooculogram, invasive blood pressure, respiratory wave, oxygen saturation, and cardiac volume as measured by VEST method. The CD-ROM also include programs to analyze polysomnography (PSG) data. The CD-ROM has values: (i) for researchers investigating clinical physiology or non-linear dynamics during sleep apnea syndrome; (ii) for engineers developing a new algorithm for the computerized analysis of PSG data related to sleep apnea syndrome; (iii) for students learning sleep physiology. PMID- 10459683 TI - Validity of sleep log compared with actigraphic sleep-wake state II. AB - Twenty-five young people (Y group), three elderly people and seven people with various sleep disorders (SD group) kept a sleep log for 2-7 days, and their wrist activity was monitored simultaneously. The sensitivity and specificity of the sleep log, and the ratio of agreement between the sleep log and actigraphic sleep wake state were calculated. The sensitivity and specificity in Y group were 87.93+/-6.49% and 96.51+/-2.37%, respectively. The sensitivity in SD group was significantly lower than in Y group. Even in Y group one-hour agreement ratios dropped during the sleep onset period. PMID- 10459684 TI - Effects of a small dose of triazolam on P300. AB - Ten healthy men (mean age, 33.9 years) participated in two experimental sessions cross-overed randomly in a double-blind manner: one with the placebo and another with 0.125 mg of triazolam (TRZ). Resting electroencephalography and event related potential under oddball paradigm were recorded before the drug administration, and 1, 2, 4, 6 and 8 h after that. P300 waveforms were analyzed by peak amplitudes and 30-ms bin data. Triazolam may cause cognitive dysfunction without general sedation or apparent sleepiness, and this effect appeared 2 h, 4 h and 6 h, most prominently 6 h, after TRZ administration. PMID- 10459685 TI - Effects of trazodone on polysomnography, blood concentration and core body temperature in healthy volunteers. AB - Polysomnography, blood concentration and core body temperature recordings were performed on 12 healthy volunteers with administration of trazodone and placebo. Trazodone increased slow wave sleep (SWS), and decreased the average, the highest and lowest core body temperature significantly compared to placebo. The blood concentration of trazodone correlated positively with amplitude (the difference between the highest and lowest temperature) and %SWS during the first period of a sleep phase divided into three periods, and negatively with the lowest temperature. The appearance time of the lowest temperature correlated negatively with %SWS. PMID- 10459686 TI - Effect of trazodone in a single dose before bedtime for sleep disorders accompanied by a depressive state: dose-finding study with no concomitant use of hypnotic agent. AB - This was the first dose-finding study of trazodone that was designed to be free of the concomitant use of hypnotics, in which the drug was administered in a single dose for sleep disorders combined with a depressive state. As a result, trazodone at the dosage of 50-100 mg/day improved sleep disorders, particularly at the 100 mg/day dosage. It was confirmed that trazodone improved sleep disorders combined with a depressive state when it was administered in a single dose before bedtime with no concomitant hypnotics. PMID- 10459687 TI - Poor recovery sleep after sleep deprivation in delayed sleep phase syndrome. AB - To clarify disturbances in sleep regulation in patients with delayed sleep phase syndrome (DSPS), we studied three patients with DSPS and seven healthy controls. Sleep propensity and melatonin rhythms after 24-h sleep deprivation were investigated under dim light condition by using the ultra-short sleep-wake schedule. The sleep propensity curves displayed clear differences between DSPS patients and the controls. During the subjective day when melatonin was not produced, recovery sleep after the sleep deprivation did not occur in DSPS patients, while recovery sleep occurred during the subjective day in controls. This suggests that DSPS may involve problems related to the homeostatic regulation of sleep after sleep deprivation. PMID- 10459688 TI - Effects of sleep deprivation: the phosphorus metabolism in the human brain measured by 31P-magnetic resonance spectroscopy. AB - Sleep deprivation (SD) has an antidepressant effect in some, but not all, patients with depression, although its biological mechanisms have not yet been characterized. We previously reported altered brain phosphorus metabolism measured by phosphorus-31 magnetic resonance spectroscopy (31P-MRS) in patients with bipolar depression. We preliminarily examined effects of SD on phosphorus metabolism in the frontal lobes of 15 normal subjects using 31P-MRS. No significant differences of membrane phospholipid metabolism, high-energy phosphate metabolism and intracellular pH were found between before and after SD in these subjects. Further studies will be necessary to elucidate the physiological mechanism of SD for depressive patients. PMID- 10459689 TI - Analysis of rapid eye movement periodicity in narcoleptics based on maximum entropy method. AB - We examined REM sleep periodicity in typical narcoleptics and patients who had shown signs of a narcoleptic tetrad without HLA-DRB1*1501/DQB1*0602 or DR2 antigens, using spectral analysis based on the maximum entropy method. The REM sleep period of typical narcoleptics showed two peaks, one at 70-90 min and one at 110-130 min at night, and a single peak at around 70-90 min during the daytime. The nocturnal REM sleep period of typical narcoleptics may be composed of several different periods, one of which corresponds to that of their daytime REM sleep. PMID- 10459690 TI - Diurnal fluctuation of sleep propensity across the menstrual cycle. AB - Most women experience sleep changes across the menstrual cycle. We applied the ultra-short sleep-wake schedule to healthy females to compare their 24-h sleep propensity rhythms in the follicular and luteal phases. The daytime (09.00-16.30 h) subjective sleepiness and the number of slow wave sleep-containing nap trials increased in the luteal phase compared to the follicular phase, but the mean sleep propensity did not change. During the periods of 17.00-00.30 h and 01.00 08.30 h there were no differences between the two phases. These results suggest that increased daytime sleepiness in the luteal phase may be related to brain mechanisms controlling slow wave sleep. PMID- 10459692 TI - The lower entrainable limit of rat circadian rhythm to sinusoidal light intensity cycles: a preliminary study. AB - The lower entrainable limit of the circadian behavioral rhythm was examined in rats exposed to sinusoidal light intensity cycles with maximum illuminance of 20 lux and the minimum of 0.01 lux. The period (T) of the light intensity cycle was initially kept at 23.5 h for 20 cycles, and then shortened to 23 h for 33 cycles. Thereafter the rats were released into constant darkness. Five out of 10 rats entrained their circadian rhythms to T = 23.5-h cycle, and they also entrained to the T = 23-h cycle. The phase angle of entrainment was almost unchanged when T was shortened from 23.5 h to 23 h. These results suggest that the T = 23-h cycle was close to the lower limit of entrainment. PMID- 10459691 TI - Double-blind test on the efficacy of methylcobalamin on sleep-wake rhythm disorders. AB - The therapeutic effect of methylcobalamin (Met-12) on sleep-wake rhythm disorders was examined in a double-blind test. In the test group which was given a large dosage, a higher percentage of improvement was found compared to the control group with a small dosage, although the difference was not significant. The test group inconsistently showed significant improvement in both the sleep-wake cycle parameters and in clinical symptoms. The tendency was for the results to show a beneficial effect of Met-12 on rhythm disorders. However, because the percentage of improvement was low and significant improvement was inconsistent, Met-12 might be considered to have a low therapeutic potency and possible use as a booster for other treatment methods of the disorders. PMID- 10459693 TI - Nurses' workload associated with 16-h night shifts on the 2-shift system. I: Comparison with the 3-shift system. AB - To assess nurses' workload with 16-h night shifts on a 2-shift system, we investigated the differences between workloads with each shift on the 2- and 3 shift systems with questionnaires on subjective symptoms and measuring heart rate and physical activity. It was found that the 2-shift nurses during the 16-h night shifts complained of fatigue less frequently and showed general decreases in heart rate and physical activity compared with the 3-shift nurses during 8-h evening and night shifts. The results suggest that a 2-h nap, an increase of staff, and a day off may reduce the nurses' workload when they have 16-h night shifts. PMID- 10459694 TI - Nurses' workload associated with 16-h night shifts. II: Effects of a nap taken during the shifts. AB - This study aimed at examining the effects on the subjective symptoms in nurses of both timing and length of a 2-h nap during a 16-h night shift. Compared to pre nap levels, sleepiness, fatigue, and dullness increased immediately after napping. Afterwards, sleepiness decreased significantly, and the other symptoms returned to the pre-nap values. The nurses' subjective symptoms after napping were not associated with the timing of the nap and post-nap fatigue lasted longer as the nap time increased (> 1.5 h). These results suggest that for effective napping during long night shifts, the nap length should be determined carefully to avoid persistent sleep inertia. PMID- 10459695 TI - Bright light treatment for sleep-wake disturbances in aged individuals with dementia. AB - Treatment using bright light exposure was carried out on six aged subjects with dementia in two nursing homes. Sleep logs were recorded by the staff. Bright light treatment was applied in the late morning every day. The %sleep in the lights-out period and/or the %wake in daytime increased in three subjects. In the other three subjects, sleep onset time was advanced. In all subjects abnormal behavior episodes around the lights-out time tended to be reduced. These results suggest that bright light treatment is effective in improving the sleep-wake disturbances of aged individuals with dementia. PMID- 10459696 TI - Effects of phototherapy in patients with delayed sleep phase syndrome. AB - Phototherapy was given to six patients with delayed sleep phase syndrome (DSPS). Polysomnography (PSG) and core body temperature were examined before and after phototherapy. Phototherapy was administered to each patient for 5 days, and this treatment not only advanced the delayed sleep phase but also delayed the time of minimum body temperature in all patients. On the PSG, decreases in total sleep time and amounts of stages 2 and REM were observed after phototherapy. These results suggest that phototherapy is effective even in the short term in advancing delays in sleep phase and time of minimum body temperature in DSPS patients. PMID- 10459697 TI - Effects of morning bright light in healthy elderly women: effects on wrist activity. AB - To investigate the effects of 8000 lux morning bright light in the elderly, home based motor activity on sleep was monitored for 5 days in 10 healthy women (mean age: 59.7 years old, range: 50-69 years old). The activity level and movement index on night 4 were significantly lower in bright light conditions, compared with the controlled condition. The activity level during the day was not significantly different between the two conditions. These results indicate that 8000 lux morning bright light improves sleep quality in healthy elderly women. PMID- 10459698 TI - Effects of morning bright light on sleep in healthy elderly women. AB - Subjective sleep feeling and polysomnography were measured in 10 elderly women to investigate the effects of 8000 lux morning bright light (BL) exposure. The profile of sleep feeling in the BL condition was better than in the control condition. The proportion of awakening time in the first one-third of night sleep decreased, and the amount of awakening time in the last one-third increased in BL condition. Daytime napping reduced in BL condition. These findings suggested the effectiveness of exposure to bright light on the improvement of sleep quality and daytime vigilance of healthy elderly women. PMID- 10459699 TI - Effects of moderately bright light on subjective evaluations in healthy elderly women. AB - Eight healthy elderly women aged 56-72 years (mean age: 67.4 years) participated in this study. They were exposed to moderately bright light (1000 lux) for 1h in the morning every 6 days in their homes. Moderately bright light improved self evaluations for alertness, mood, motivation, happiness, refreshment, concentration and appetite after the second exposure to the light. Sleep maintenance, anxiety and integrated sleep feeling also improved after the fourth exposure to the light. The phase of rectal temperature rhythm did not change in a moderately bright light condition. These results suggest that bright light influences sleep quality and self evaluations without changing the biological rhythm phase. PMID- 10459700 TI - The hypnotic effects of melatonin treatment on diurnal sleep in humans. AB - This study investigated the hypnotic effects of 10 mg melatonin and placebo, which were administered at 10.00 h, according to a single-blind crossover design, on an 8-h diurnal sleep from 11.00 to 19.00 h, following a full night of sleep. The subjects were six healthy male students, each of whom underwent polysomnography and rectal temperature monitoring. Melatonin treatment significantly increased total sleep time in diurnal sleep (403.2+/-SD 72.8 min and 258.5+/-118.3 min, P<0.001). As to changes in rectal temperature during diurnal sleep, however, there were no significant differences between the melatonin and placebo conditions. Thus, these results indicated that melatonin administered at 10.00 h had direct hypnotic effects on diurnal sleep. PMID- 10459701 TI - A newly developed assay for melatonin using cells expressing human mel-1a receptor. AB - We have developed a radioreceptor binding assay (RRA) method for melatonin using membranes from Chinese hamster ovary cells that can stably express human mel-1a receptors. We measured melatonin levels in plasma samples collected every 4h for 24h using the RRA and radioimmunoassay (RIA) methods, simultaneously. There was a statistically significant correlation between the melatonin levels measured by the two methods, this newly developed method providing a sensitive bioassay. As it is possible to circumvent the cross-reactivity usually occurring in the RIA method, this method may be an important tool for detecting bioactive substances relative to the mel-1a receptor. PMID- 10459702 TI - Amplitude reduction of plasma melatonin rhythm in association with an internal desynchronization in a subject with non-24-hour sleep-wake syndrome. AB - The plasma melatonin rhythm was measured longitudinally in a subject with non-24 h sleep-wake syndrome, and the amplitude and area under the curve (AUC) of the melatonin rhythm were investigated in relation to the sleep-wake cycle. When the melatonin rhythm and sleep-wake rhythm were internally desynchronized, the amplitude and the AUC were reduced significantly. These parameters were not influenced by external melatonin administration. These results suggest that a causal relationship between the reduction of circadian oscillation and internal desynchronization exists in this subject. PMID- 10459703 TI - Correlation between the circadian sleep propensity rhythm and hormonal rhythms under ultra-short sleep-wake cycle. AB - The aim of this study was to clarify effects of hormonal and temperature rhythms on circadian fluctuations of sleep propensity. Ten healthy females underwent 24-h sleep deprivation and entered the circadian sleep propensity assessment setting under the ultra-short sleep-wake schedule. During the experiment, sleep propensity rhythm, rectal temperature, and 24-h serum hormone profiles (melatonin, cortisol and thyroid-stimulating hormone) were investigated. The circadian sleep propensity rhythms had two apparent peaks (afternoon and nocturnal peaks) and a trough (nocturnal sleep gate). The timings of the nocturnal sleep gate and the nocturnal peak were correlated exclusively with temperature and melatonin rhythms (P < 0.05), while that of the afternoon peak was significantly correlated with habitual wake time and melatonin rhythm. These results indicate that the circadian sleep propensity rhythm is influenced not only by the circadian pacemaker, but also by sleep habit. PMID- 10459704 TI - Re-entrainment of circadian rhythm of plasma melatonin on an 8-h eastward flight. AB - To estimate the process of re-entrainment we measured the melatonin rhythm on an eastward flight. After the baseline study, 24-hour blood sampling of six male subjects was done on the first and fifth days. During the daytime the subjects were exposed to natural zeitgeber outdoors every day except the blood sampling day. They were analyzed with an illuminometer when under the bright light condition. Four of the six subjects showed orthodromic re-entrainment, another subject showed antidromic re-entrainment, and the other subject kept the baseline pattern of plasma melatonin. The rate of re-entrainment in orthodromic re entrainment was about 55 min per day. Measuring the circadian rhythm of plasma melatonin has clarified the interindividual re-entrainment difference. PMID- 10459705 TI - Comparative studies on sleep disturbance in the elderly based on questionnaire assessments in 1983 and 1996. AB - We made an attempt to compare the complaints about sleep disturbance in the elderly based on the questionnaires using self-rating scales carried out in 1983 and 1996. It was noted that the score of awakening frequency from the course of sleep was the highest of all the items in men and women in both the 1983 and 1996 inquiries. The scores of insomnia nights per week and the difficulty in falling asleep were slightly higher in women than in men in both inquiries. PMID- 10459706 TI - What prevents Japan from establishing a clinical service of sleep medicine: experience gained at a local hospital. AB - We opened a mini-sleep disorders clinic in a local general hospital in rural Japan, and have been operating it for more than 3 years. We encountered many difficulties during this period when we tried to provide proper clinical service. Problems ranged from lack of sleep specialists and polysomnography technicians, shortage of funding, non-existence of an educational system to obtain comprehensive knowledge about sleep medicine, and also lack of cooperation among different departments necessary for maintaining clinical service. Through our experience, we analyzed various factors that prevent Japan from establishing a clinical service of sleep medicine. PMID- 10459707 TI - Demography of sleep disturbances associated with circadian rhythm disorders in Japan. AB - To clarify the demography of delayed sleep phase syndrome (DSPS), non-24-h sleep wake syndrome, and irregular sleep-wake pattern in Japan, a cross-sectional nationwide epidemiological survey was conducted. 1525 adults (age: 15-59 years) were randomly sampled from telephone directories, and they received screening questions over the phone. Persons who were suspected of having the disorders were requested to fill out the second questionnaire, and asked to keep a sleep log for 4 weeks. Diagnoses were made according to the International Classification of Sleep Disorders criteria. As a result, the prevalence of DSPS was estimated to be 0.13%. PMID- 10459708 TI - Lifestyles and sleep disorders among the Japanese adult population. AB - To clarify the effects of daily stress, habitual exercise, drinking and smoking on the prevalence of sleep disorders, we selected 4000 residents (> or =20 years) in Japan by stratified random sampling and carried out structured interviews (response rate 75.8%). Frequencies of sleep disorders (difficulty initiating sleep: DIS; difficulty maintaining sleep: DMS; early morning awakening and hypnotic medication use) were treated as dependent variables. Significant effects of stress were prevalent in all sleep disorders. Habitual exercise had significant negative association with DIS and DMS. Drinking and smoking did not affect sleep disorders. PMID- 10459709 TI - Effects of volitional lifestyle on rest-activity cycle in the aged. AB - The present study investigated the relationship among volitional lifestyle, activity and sleep in the aged. We selected 28 subjects over 65 years of age to survey volition to lifestyle. High-volitional group (14 Ss, mean age: 74.1 years, seven males and seven females) and low-volitional group (14 Ss, mean age: 73.0 years, seven males and seven females) were identified by the scale of self confidence and the Philadelphia Geriatric Center morale scale. Wrist activities were monitored by Actigraph for 10 or 14 consecutive days. The subjects recorded sleep logs by themselves for those days. It was found that high volitional aged individuals had a relatively well structured sleep in comparison with low volitional aged individuals, and that most of them took a clock dependent habitual nap around 14.00 h. On the other hand, low volitional aged individuals had a relatively poor structured sleep, and took a nap time-dependently after 8 h from the morning rising time. These results suggest that high volitional aged individuals build a short nap into their well organized sleep-life habits. PMID- 10459710 TI - Effects of a daytime nap in the aged. AB - This study evaluated the effects of the daytime nap on performance, mood and physiological measures in aged individuals Participants were six healthy aged persons (M=72.2 years old) who habitually napped in the afternoon three or more times a week. They participated under two conditions with an interval of 1 week. In the nap condition, the subjects went to bed at 13:00 h and slept for 30 min. In the rest condition, they just watched television. In both conditions, electroencephalogram (EEG), blood pressure, mood and performance were measured before and after a nap or rest. The daytime nap improved performance, decreased subjective sleepiness and fatigue, and attenuated EEG alpha band activity. Moreover, following a nap diastolic blood pressure significantly decreased. These findings suggest that a habitual daytime nap helps aged individuals to maintain their daytime physiological, psychological and behavioral arousal at an adequate level. PMID- 10459711 TI - Systematic treatment for nocturnal urinary frequency following a sleep micturition chart. AB - Seven patients complaining of nocturnal urinary frequency were treated following a sleep-micturition chart. By clinical analysis of the data, the causes of nocturnal urinary frequency were divided into three diagnostic categories: a small bladder capacity group, a sleep disorder group and a large nocturnal urinary volume group. The three groups were treated by anticholinergic agents, sleeping pills and restriction of water intake at night respectively. With the administration of anti-cholinergic agents or sleeping pills nocturnal bladder capacity increased. By restriction of water intake at night nocturnal urinary volume decreased. In all patients nocturnal urinary frequency decreased and sleep efficiency was improved. PMID- 10459712 TI - A case of nocturnal polyuria in olivopontocerebellar atrophy. AB - We report a case of olivopontocerebellar atrophy without sleep apnea syndrome who presented nocturnal polyuria. It is considered that a disturbance in the circadian rhythm for arginine vasopressin secretion due to degeneration of suprachiasmatic nuclei and marked increase in the secretion of atrial natriuretic peptide due to abnormal diurnal variation in blood pressure may be involved in the mechanism of nocturnal polyuria. PMID- 10459713 TI - Treatment of periodic leg movement disorder and restless leg syndrome with talipexole. AB - In order to investigate the bedtime dose of talipexole, a D2 and alpha2 stimulant, on patients with periodic leg movement disorder (PLMD) and restless leg syndrome (RLS), we made a comparison of polysomnographic findings and subjective symptom ratings before and during 4 weeks of the treatment on five cases with RLS and PLMD. A significant decrease in both the frequencies of periodic leg movements and subjective symptom ratings and significant improvement of sleep composition were recognized during the treatment. We speculate that the combination of the agonistic action of D2 and alpha2 receptor with the drug might not only suppress PLMD and RLS but also improve sleep quality. PMID- 10459714 TI - Disappearance of rhythmic involuntary movements during sleep in a case of olivopontocerebellar atrophy. AB - We report on a 54-year-old woman with an 8 or so year history of olivopontocerebellar atrophy associated with the rhythmic involuntary movements of the left upper and lower limbs, and cervical region. Surface electromyogram of the left upper limb revealed rhythmicity (about 3 Hz) and reciprocity between antagonistic muscles, which disappeared on polysomnography at all sleep stages including rapid eye movement sleep without atonia. These were characterized by the co-existence of rhythmic skeletal myoclonus and parkinsonian tremor. These findings suggest that a disturbance of the striatonigral system as well as the dentato-rubro-olivary circuit may be involved in these movements. It also seems that their fate is dependent on the level of wakefulness and that the ascending reticular activating system also plays a role in the development of these movements. PMID- 10459715 TI - A case of neuroleptic-induced unilateral akathisia with periodic limb movements in the opposite side during sleep. AB - We report on a patient with schizoaffective disorder who developed unilateral akathisia. This is the first case report of a patient with neuroleptic-induced unilateral akathisia on whom an all-night polysomnogram was recorded. On the polysomnogram we observed right side periodic limb movements (PLM) with left side unilateral akathisia, and after her akathisia disappeared, the PLM also disappeared. Brain MRI findings and neurological findings were within normal limits. The pathogenetic lesion causing akathisia could not be elucidated. PMID- 10459716 TI - Two cases of HLA-DR2-negative hypersomnia manifesting sleep-onset rapid eye movement periods in the multiple sleep latency test. AB - We encountered two cases expressing excessive daytime sleepiness (EDS) and manifesting two or more sleep-onset rapid eye movement (REM) periods in the multiple sleep latency test. Unbearable daytime sleepiness occurred abruptly, which usually led to short-lasting naps, after which the patients felt refreshed. The EDS was successfully reduced by treatment with methylphenidate. In spite of these features similar to narcolepsy, these cases of REM hypersomnia did not present cataplexy or other auxiliary symptoms of narcolepsy, and, furthermore, the class-II human leukocyte antigen DR2 appeared to be negative. PMID- 10459717 TI - Intensity of snoring in patients with sleep-related breathing disorders. AB - The intensity of snoring was measured in 73 patients with snoring or sleep apnea using a noise meter. The mean intensity of snoring at 50 cm in front of the mouth was 61.7 dB in the supine position and 53.7 dB in the lateral position. There existed a definite correlation between the logarithmic transformation of the intra-esophageal pressure amplitude and the intensity of snoring in the supine and lateral decubitus positions. These findings suggest that the intensity of snoring may be a useful index for sleep-related breathing disorders. PMID- 10459718 TI - Epworth Sleepiness Scale and sleep studies in patients with obstructive sleep apnea syndrome. AB - Excessive daytime sleepiness (EDS) is the major symptom of patients with obstructive sleep apnea syndrome (OSAS). In this study, we examined the relationship between subjective EDS scored with the Epworth Sleepiness Scale (ESS), objective EDS measured with the multiple sleep latency test (MSLT) and sleep variables evaluated with polysomnography for patients with OSAS. Subjects were 10 patients (51.7+/-19.0 years old). The average ESS and MSLT scores were 10.6+/-5.6 and 7.7+/-5.6, respectively. There was no significant relationship between ESS and MSLT. The Multiple Sleep Latency Test had a significant negative relationship with the number of awakenings and the apnea/hypopnea index. No relationship was found between nocturnal hypoxia and either ESS or MSLT. Our findings suggest that objective EDS in OSAS is related with fragmentation of sleep, and that several patients are not aware of their EDS. PMID- 10459719 TI - Treatment of obstructive sleep apnea syndrome with a Kampo-formula, San'o-shashin to: a case report. AB - The following describes a 76-year-old male with obstructive sleep apnea syndrome successfully treated with a Kampo-formula, San'o-shashin-to (Formula medicamentorum tres ad dispellendi cordis). Polysomnography, performed before and after administration of San'o-shashin-to, revealed that the apnea index decreased from 11.1 events/hour to 4.1 events/hour, and that the apnea plus hypopnea index decreased from 18.4 events/hour to 10.7 events/hour. The patient was normo-weight (body mass index: 20.4 kg/m2), and events of sleep apnea and hypopnea were mostly noted during a non-rapid eye movement sleep. It is possible that San'o-shashin-to has some alleviating effects on the upper airway resistance during sleep. PMID- 10459720 TI - Frequent breathing-related electroencephalogram arousals in four patients with mild obstructive sleep apneas. AB - We report cases of four patients with mild obstructive sleep apnea syndrome (OSAS) with frequent breathing-related electroencephalogram (EEG) arousals which led to excessive daytime sleepiness. In spite of a relatively low apnea hypopnea index (AHI), sleep was disrupted by frequent EEG arousals associated with respiratory effort as observed in upper airway resistance syndrome. The effects of sleep stage and sleep position on EEG arousals were also investigated. We consider that AHI alone is not a sufficient index to assess severity of OSAS, and it is very important to examine microarousals by the alteration of esophageal pressure in addition to the effect of sleep position. PMID- 10459721 TI - Multiple cardiovascular risk factors in obstructive sleep apnea syndrome patients and an attempt at lifestyle modification using telemedicine-based education. AB - Severe obstructive sleep apnea syndrome (OSAS) is a typical 'lifestyle-related disease' characterized by a high incidence of cardiovascular risk factors, such as obesity, smoking, hypertriglyceridemia, and diabetes mellitus. Patients with OSAS tend to have eating disorders as a result of efforts to overcome the intolerable sleepiness. Treatment of OSAS should therefore aim to improve the lifestyle through encouraging weight reduction, physical activity increase, and tobacco avoidance, in addition to direct therapy such as continuous positive airway pressure for upper airway obstruction. The telemedicine system we developed was considered to be effective for providing home-based education on nutrition and exercise aimed at correcting multiple risk factors in OSAS patients PMID- 10459722 TI - Effects of prone and supine position on sleep characteristics in preterm infants. AB - The purpose of this study was to address the influence of sleep position on sleep characteristics in preterm infants. We studied 16 infants at a mean post conceptional age of 36.5 weeks. Infants were successfully recorded with videopolysomnograph in the supine and prone position. Between the two positions, there were no significant differences in percentage of active sleep and quiet sleep (QS), the occurrence of arousal, and the incidence of apnea. The first QS after the feeding was longer in the prone position. The sleep position could affect sleep characteristics but not respiratory characteristics in preterm infants. PMID- 10459723 TI - Polysomnographic and urodynamic changes in a case of obstructive sleep apnea syndrome with enuresis. AB - A 53-year-old female patient with obstructive sleep apnea syndrome was reported. She had complained of enuresis as well as a 15-year history of snoring, but she had no complaint of sleep and awake disturbance. Polysomnographic study showed repeated obstructive apnea and hypopnea with an apnea/hypopnea index of 52.6, and severe oxygen desaturation during sleep. On cystometography during sleep, the changing amplitude of the spike wave corresponds to the changes of respiratory efforts against a closed upper airway. The patient was treated successfully with imipramine and acetazolamide for the obstructive sleep apnea and enuresis. Apnea/hypopnea index, nocturnal oxygen desaturation, and sleep architecture were improved, and enuresis completely disappeared. Cystometrography during sleep showed that the average amplitude of the spike wave tended to be low. Percentage urinary volume during sleep compared with 24 h volume was significantly reduced. We considered that the enuresis was mainly related to increased intra-abdominal pressure produced by respiratory efforts and enhanced nocturnal urine production. PMID- 10459724 TI - Clinical efficacy and indication of acetazolamide treatment on sleep apnea syndrome. AB - The efficacy and indication of acetazolamide treatment on patients with sleep apnea syndrome (SAS) were discussed from assessing the changes of polysomnographic findings with the treatment in 75 SAS patients. For the patients as a whole, respiratory disorder variables improved significantly during the treatment. However, the number of acetazolamide treatment responders who showed a decrease of apnea hypopnea index (AHI) to 50% or less of the pretreatment value numbered only 34 (45.3%). The lower values of body mass index and AHI in the responder group indicated that monotherapy with acetazolamide is the treatment choice only for mild SAS cases without obesity. However, combined treatment with acetazolamide and uvulopalatopharyngoplasty was thought to be beneficial for severe cases. PMID- 10459725 TI - Clinical efficacy of prosthetic mandibular advancement on obstructive sleep apnea syndrome. AB - In order to clarify the efficacy and indication of prosthetic mandibular advancement (PMA) on obstructive sleep apnea syndrome (OSAS), we made a comparison of both the polysomnographic findings and the upper airway configuration between before and during PMA on 19 OSAS patients. During PMA, there was a significant decreased apnea hypopnea index compared to before treatment. The changes in magnetic resonance imaging of the upper airway during sleep with PMA indicated that the treatment is regarded to be the first choice for OSAS patients with glossopharyngeal obstruction. In addition to that, the treatment might be considered for use in velopharyngeal obstruction. PMID- 10459726 TI - Evaluation of the auto continuous positive airway pressure efficacy by upper airway pressure measurement. AB - The efficacy of auto continuous positive airway pressure (CPAP) was prospectively evaluated in 11 obstructive sleep apnea cases by the upper airway pressure measurement. Their apnea plus hypopnea index decreased from 68.3+/-20.2/h to 0.5+/-0.9/h during manually adjusted and 9.6+/-14.5/h during auto CPAP The intraesophageal pressure decreased from -88.0+/-34.0 cm H2O to 11.9+/-7.2 cm H2O during manually adjusted and -26.0+/-13.7 cm H2O during auto CPAP. The manually adjusted CPAP level was 9.1+/-1.7 cm H2O and the maximum pressure of the auto CPAP was 9.4+/-2.0 cm H2O. Auto CPAP could automatically detect the optimal CPAP level. However, the efficacy of the auto CPAP was less than that of the manually adjusted CPAP. PMID- 10459727 TI - Clinical characteristics of upper airway resistance syndrome. AB - Polysomnographic findings and clinical symptoms were investigated in 14 cases of upper airway resistance syndrome. The mean scores of the Epworth sleepiness scale and self-rating depression scale in eight cases were 13.5 and 38.6, respectively. The mean sleep latency of the multiple sleep latency test in four cases was 10.2 min. Seven cases were treated with continuous positive airway pressure (CPAP), and one with hormone replacement therapy. The most common symptom was daytime sleepiness. Five cases had hypertension. CPAP reduced increasing negative esophageal pressure (Pes) and frequency of EEG arousals, and improved hypertension in one case. Hormone replacement therapy ameliorated increasing negative Pes and clinical symptoms. PMID- 10459728 TI - Monoclonal antibody-directed cytotoxic therapy: potential in malignant diseases of aging. AB - The advent of monoclonal antibodies has allowed the development of tumour directed therapies utilising antibody-dependent effector mechanisms and immunoconjugates (e.g. drug, isotope and toxin coupled antibodies) against human malignancies. Preclinical studies in mouse tumour models have been most impressive and have led to numerous clinical trials. Whereas the majority of these phase I/II trials have been less impressive, a few trials have shown efficacy in highly pre-treated refractory patients and have led to phase III trials. The therapeutic monoclonal antibodies examined in these trials will become clinically available in the near future. In this review, various methods of utilising antibody-directed anticancer strategies are presented, with emphasis on recent advances in the field. The advantages and disadvantages of these methods together with the role of antibody-directed therapeutics in cancer management are discussed. PMID- 10459729 TI - Drug-induced cognitive impairment in the elderly. AB - Elderly people are more likely than younger patients to develop cognitive impairment as a result of taking medications. This reflects age- and disease associated changes in brain neurochemistry and drug handling. Delirium (acute confusional state) is the cognitive disturbance most clearly associated with drug toxicity, but dementia has also been reported. The aetiology of cognitive impairment is commonly multifactorial, and it may be difficult to firmly establish a causal role for an individual medication. In studies of elderly hospital patients, drugs have been reported as the cause of delirium in 11 to 30% of cases. Medication toxicity occurs in 2 to 12% of patients presenting with suspected dementia. In some cases CNS toxicity occurs in a dose-dependent manner, often as a result of interference with neurotransmitter function. Drug-induced delirium can also occur as an idiosyncratic complication. Finally, delirium may occur secondary to iatrogenic complications of drug use. Almost any drug can cause delirium, especially in a vulnerable patient. Impaired cholinergic neurotransmission has been implicated in the pathogenesis of delirium and of Alzheimer's disease. Anticholinergic medications are important causes of acute and chronic confusional states. Nevertheless, polypharmacy with anticholinergic compounds is common, especially in nursing home residents. Recent studies have suggested that the total burden of anticholinergic drugs may determine development of delirium rather than any single agent. Also, anticholinergic effects have been identified in many drugs other than those classically thought of as having major anticholinergic effects. Psychoactive drugs are important causes of delirium. Narcotic agents are among the most important causes of delirium in postoperative patients. Long-acting benzodiazepines are the commonest drugs to cause or exacerbate dementia. Delirium was a major complication of treatment with tricyclic antidepressants but seems less common with newer agents. Anticonvulsants can cause delirium and dementia. Drug-induced confusion with nonpsychoactive drugs is often idiosyncratic in nature, and the diagnosis is easily missed unless clinicians maintain a high index of suspicion. Histamine H2 receptor antagonists, cardiac medications such as digoxin and beta-blockers, corticosteroids, non-steroidal anti-inflammatory agents and antibiotics can all cause acute, and, less commonly, chronic confusion. Drug-induced confusion can be prevented by avoiding polypharmacy and adhering to the saying 'start low and go slow'. Special care is needed when prescribing for people with cognitive impairment. Early diagnosis of drug-induced confusion, and withdrawal of the offending agent or agents is essential. PMID- 10459730 TI - Aspirin and heparin in acute ischaemic stroke in older patients. AB - Over four-fifths of all strokes are due to thrombotic or embolic occlusion of cerebral arteries. There is a strong rationale for considering antithrombotic therapy for the treatment of patients with acute ischaemic stroke. Antiplatelet therapy with 150 to 300 mg of aspirin (acetylsalicylic acid) started within the first 48 hours of an ischaemic stroke improves patient outcome in the short and long term, with a low risk of adverse effects. Anticoagulants such as heparin may reduce the risk of developing deep venous thrombosis and pulmonary embolism in patients with stroke, but randomised controlled trials have demonstrated a significant and dose-dependent risk of intracranial haemorrhage. The routine use of parenteral anticoagulants, including unfractionated heparin, low-molecular weight heparins and heparinoids in the acute phase of ischaemic stroke is not associated with any net short or long term benefit. Aspirin is, therefore, the antithrombotic drug of choice in the treatment of acute ischaemic stroke. PMID- 10459731 TI - Health care rationing in the aged: ethical and clinical perspectives. AB - This article provides an ethical analysis of the question of whether aged patients' access to health care should be less than, the same or greater than, the access younger patients enjoy, when economic resources are limited. This topic is being urgently considered in the US because managed care is becoming more common and brings with it new challenges to traditional medical ethics, and because the prevalence of the aged is increasing as is the number of patients with Alzheimer's disease (AD). It is also critical throughout the world because new findings suggest that the progression of AD may be retarded or even reversed by providing patients with enriched interpersonal environments. If these findings are valid, it would be inhumane to not consider providing these resources to patients with AD, since these gains would be so meaningful and substantial. Deontological and consequential values influencing this question are presented and evaluated. The theories of Veatch, Callahan and Daniels in regard to allocating health care to the aged are discussed. It is argued that 2 subgroups of aged patients, the isolated and demented, are among the patients worst off because the capacity to enjoy meaningful relationships with others supercedes all others and both groups of patients have lost this capacity. We assert that, on the basis of the principle of justice according to need, these 2 groups of patients' exceptional needs should be prioritised. We then raise the question of whether the majority of the population would be willing to provide these isolated and demented aged patients this care if the new findings proved valid. We conclude that, in light of many people's fear of growing old and dying, and some peoples bias against the aged (particularly in the US), willingness to provide the necessary resources is open to debate. Finally, we provide specific examples of the kinds of interventions which might be optimal for each group of patients. For patients who are cognitively unimpaired, this might be providing home care so that they could remain closer to and in contact with their loved ones. For patients who are cognitively impaired, this might be providing interpersonal support when these patients begin to lose control, rather than applying restraints or using psychotropic medication. PMID- 10459733 TI - Valrubicin. AB - Valrubicin (AD-32) is an N-trifluoroacetyl, 14-valerate derivative of the anthracycline doxorubicin. It has antineoplastic activity which probably results from interference with nucleic acid metabolism by the drug. Valrubicin entered individual cells more rapidly than doxorubicin in vitro. When valrubicin was administered intravesically to patients with bladder cancer, cytotoxic concentrations of the drug penetrated the superficial muscle layer of the bladder. Complete response rates were 18 and 29% in patients with carcinoma in situ of the bladder which was refractory to intravesical BCG in 2 non-comparative trials of prophylactic intravesical valrubicin. In patients with recurrent superficial papillary tumours, the complete response rate was 46%. Adverse events were generally transient in patients who received intravesical valrubicin. Bladder irritation occurred in 88% of patients. Systemic absorption of intravesically administered valrubicin was minimal. Accordingly, systemic adverse events generally occurred in < or =5% of patients. Valrubicin was less toxic to chick embryos and haematopoietic stem cells in vitro and produced a lower incidence of cardiotoxicity in rabbits, compared with doxorubicin. PMID- 10459732 TI - Photodynamic therapy: a review. AB - Photodynamic therapy (PDT) of malignant tumours is a new technique for treating cancers. After intravenous injection, a photosensitiser is selectively retained by the tumour cells so after time there is more sensitiser in the tumour than in the normal adjacent tissue. The photosensitiser must be able to absorb the wavelength of light being delivered to it, and the amount of light getting to the photosensitiser depends on the characteristics of the tissue it passes through. When exposed to light with the proper wavelength, the sensitiser produces an activated oxygen species, singlet oxygen, that oxidises critical elements of neoplastic cells. Because there is less sensitiser in the adjacent normal tissue, less reaction occurs to it. Since this is an entirely different process, the use of chemotherapy, ionising radiation or surgery does not preclude the use of PDT. Also, unlike ionising irradiation, repeated injections and treatments can be made indefinitely. Different molecules and atoms absorb different wavelengths of energy. Since the light energy must be absorbed to start the photochemical reaction, the absorption spectrum of the photosensitiser determines the wavelength used to initiate the reaction. However, this can be qualified by the tissue the light has to travel through to get to the photosensitiser. The photosensitiser porfimer sodium has a peak absorption in the area of 405 nm (blue violet) and a much lower absorption peak at 630 nm (red). However, because the longer red wavelength penetrates tissue deeper than 405 nm, we use the red wavelength, usually delivered from a laser system. This permits coupling of the red light beam to quartz fibres which can then be used with modifications to treat external surface tumours, inserted interstitially directly into large tumours, passed though any endoscope to treat intraluminal tumours, or inserted behind the retina to treat tumours of the retina. Twenty years after the pioneering work of Dr. Thomas Doherty, the US Food and Drug Administration (FDA) has approved the use of porfimer sodium for photodynamic therapy of endobronchial and oesophageal tumours. Research continues towards approval for management of skin cancers and metastatic cutaneous and subcutaneous breast cancers. The realisation that one of the mechanisms of photodynamic therapy is thrombosis of vessels led to the development of verteporfin to treat macular degeneration. Multiple other areas are being investigated as well as new photosensitisers. Photodynamic therapy is an entirely new treatment modality and its development can be likened to that of the discovery of antibiotics. This is just the beginning, and its possible uses are only limited by the imagination. PMID- 10459734 TI - Schizophrenia: is it time to replace the term? AB - The attitudes of Japanese psychiatrists toward their patients who suffer from schizophrenia were investigated. We were concerned specifically with whether the psychiatrists inform their patients of the suspected diagnosis. We discuss how the term 'schizophrenia' may influence a psychiatrist's decision to inform his patients of the diagnosis. A self-reported questionnaire was distributed to 150 executive board members of the Japanese Society of Psychiatry and Neurology and analysis of the data obtained from 110 respondents was carried out. The results showed that the concepts that psychiatrists use when they give a diagnosis of schizophrenia vary considerably. Fifty-nine per cent of the respondents informed their patients of a diagnosis of schizophrenia on a case-by-case basis, while 37% informed only the patients' families. A tree analysis showed that the most important predictors for informing the patients of the diagnosis were assumptions about the public image of schizophrenia and a negative impression of the term schizophrenia, translated as 'Seishin Bunretsu Byou' in Japanese. The results revealed that the Japanese term for schizophrenia influences a psychiatrist's decision to inform patients of the diagnosis and that, by changing the term to a less stigmatized one, the disclosure of information about schizophrenia to patients would be promoted. PMID- 10459735 TI - Symptom structure and psychiatric comorbidity of combat-related post-traumatic stress disorder. AB - The placement of the diagnostic category of post-traumatic stress disorder (PTSD) among anxiety disorders reflects the recognition that anxiety is a predominant reaction to trauma. Indeed, the symptoms of PTSD overlap considerably with those of other anxiety disorders. The nosological criteria render PTSD as quite a heterogeneous diagnosis. Two individuals with no common symptoms can be diagnosed as having PTSD. In this report we provide information on the phenomenology and psychiatric comorbidity in a sample of 33 patients with combat-related PTSD. The finding of clinical heterogeneity in subjects with combat-related PTSD and the therapy implications are discussed. PMID- 10459736 TI - Regional cerebral blood flow abnormalities in chronic solvent abusers. AB - This study aimed to reveal regional abnormalities of cerebral blood flow (CBF) and their relation to amotivational syndrome which causes poor social prognosis in solvent abusers Sixteen chronic solvent abusers (12 males and four females) along with five normal subjects underwent single photon emission computed tomography with N-isopropyl-p[123I]iodoamphetamine. The patients received a clinical evaluation with the Scale for the Assessment of Negative Symptoms. Using a semiquantitative method (normalized by the parietal cortex count), patients showed a statistically significant decrease in regional cerebral blood flow (rCBF) in the bilateral prefrontal cortices (P<0.01). In addition, the severity of hypoperfusion in the bilateral prefrontal cortices was related to the degree of severity of the avolition-apathy scale on SANS (left; P<0.05, right; P<0.01) even after excluding the effect of antipsychotics. These results suggest that rCBF abnormalities, especially in the prefrontal cortex, develop in chronic solvent abusers, and that this frontal hypoperfusion may be a biological basis of amotivational syndrome. PMID- 10459737 TI - Grief reactions treated in a psychiatric out-patient clinic. AB - Grief reactions to the death of a close relative are not commonly treated in psychiatric out-patient clinics in Japan. However, recently our clinic has had a chance to see such cases. I analyzed 11 cases of grief reactions (7 females, 4 males). The first part of this paper focuses on the distinction between normal and abnormal grief reactions, and an attempt was made to classify grief reactions in the nosology. Following this, three factors were specified as enhancing vulnerability to bereavement: (1) sex (female), (2) age (middle-aged), and (3) cause of death (sudden death). The second part focuses on the dependency seen in the subjects, which was their most marked personality traits. Finally, vertical splitting as defined by Kohut and Kasahara was discussed as a defense mechanism in the crisis of bereavement. The bereaved person preserves a positive image of the deceased and his or herself by vertical splitting, thereby eliminating negative experiences and feelings (denial). This manifested itself in a strong feeling of solidarity with the deceased (identification). The treatment of the subjects followed a fairly good course, but five patients required more than 1 year. Four patients visited the clinic after the termination of their treatment because of a relapse of their depression. PMID- 10459738 TI - Weight and shape concerns and dieting behaviours among pre-adolescents and adolescents in Japan. AB - The attitudes towards body weight and shape, desire for thinness and dieting behaviours were investigated in pre-adolescent and adolescent girls and boys (547 elementary school students, 615 junior high school students, and 470 senior high school students) aged 10-17 years in Osaka Prefecture, Japan, by a self-report questionnaire. Forty-eight per cent of 10-year-old females and 84% of 17-year-old females categorized themselves as 'fat' or 'too fat'. The fear of weight gain and desire for thinness was reported in 35% and 51% of 10-year-old girls, respectively, and increased progressively with ageing to 79% and 87% of 17-year old girls. In contrast, these were reported by 20-30% of boys in the corresponding age groups. Some practices to be slim were found in 22% of the 10 year-old girls, and increased to 37% among the 17-year-old girls, whereas they were found in around 20% of the boys at each age. The practices to be slim were found in 32% of the girls who were 85-90% of the standard body weight (SBW) and in 14% of the girls less than 85% of the SBW. These results suggest that significant concerns about weight and shape and dieting behaviours are present in young Japanese girls and increase progressively with age. These results are compatible with those in Western society. PMID- 10459739 TI - A profile analysis of personality disorders: beyond multiple diagnoses. AB - The authors applied cluster analysis and multidimensional scaling to the analyses of 59 patients with personality pathology. Cluster analysis yielded eight typologies of patients: detached, anankastic, phobic, dramatic, erratic, emotional, milder emotional, and masochistic negativistic. Multidimensional scaling identified the dimensions of classifying patients: anxious rumination versus behavioural acting out, overall severity of personality pathology, and assertiveness versus withdrawal. Considering the distinction between personality disorder (dysfunctional personality) and abnormal personality (extreme personality), the following changes in current classification system are proposed: use of a hierarchy and exclusion criteria in a categorical-type model or use of a personality profile in a dimensional-trait model, in either case, with a dimensional rating for severity of psychopathology to define personality 'disorder'. PMID- 10459740 TI - Application of the Zung self-rating depression scale to patients before and after introduction to haemodialysis. AB - In the present study we applied the Zung self-rating depression scale (SDS) to 19 renal failure patients who were introduced to haemodialysis in the Nagoya Daini Red Cross Hospital, Nagoya, Japan. The patients were divided into two groups: the emergent introduction (EI) group, who underwent unanticipated and sudden introduction to haemodialysis, and the ordinary introduction (OI) group, who experienced a more systematic introduction to haemodialysis following recommendation by medical specialists. The patients' Zung SDS responses were collected twice, just before and 2 weeks after haemodialysis introduction. The total SDS score of the EI group was significantly higher than that of the OI group, both before and after haemodialysis introduction. The total SDS scores for the EI and OI groups were significantly reduced after haemodialysis introduction. The SDS scores for the EI group were significantly higher in the mood of depression and cognitive symptoms categories, both before and after haemodyalysis introduction. Before introduction, SDS scores of the EI group were significantly higher in the categories of motor and vegetative symptoms. The SDS scores for vegetative symptoms in the EI group significantly decreased after introduction to haemodialysis. These results suggest that haemodialytic excretion of uremic toxins helps to reduce SDS scores. PMID- 10459741 TI - A polydiagnostic study of depressive disorders according to DSM-IV and 23 classical diagnostic systems. AB - The classification of mood disorders is one of the most highly debated topics in modern psychiatry. The introduction of DSM-III (and its followers) has set a new standard in this controversy but little empirical evidence is available as to how the various classical diagnostic categories of mood disorders by Kraepelin, Schneider, Leonhard, Hamilton, Kielholz, Winokur and others compare with this new standard. The Intensive Prospective Study arm of the Group for Longitudinal Affective Disorders Study has studied a broad spectrum of mood disorders in 23 participating centres from all over Japan with a polydiagnostic semistructured interview called Comprehensive Assessment List of Affective disorders. In this paper we examined how the various classical diagnostic systems of depressive disorders correspond to the DSM-IV diagnoses, and found the following: (1) The classical 'neurotic' or 'psychogenic' depressions are diagnosed as major depression and not as dysthymia in DSM-IV; although dysthymia was dubbed as 'depressive neurosis' in DSM-III, its criteria were not true to the traditional usage of the term. Viewed from the other side of the coin, DSM-IV can be said to stand in the unitary tradition. (2) Some of the classical diagnostic categories such as Schneider's depressive psychopathy and Klein's acute dysphoria as well as modern ones such as Akiskal's subaffective dysthymia and Angst's recurrent brief depression were rarely seen in traditional psychiatric treatment settings. (3) Comparisons of the unique diagnostic systems such as those by Leonhard, Winokur and Berner warrant further studies on their validity. PMID- 10459742 TI - Aldehyde dehydrogenase genotypes and male alcohol use disorders: a case-control study in Khon Kaen, north-east Thailand. AB - A genetic epidemiological case-control study on aldehyde dehydrogenase 2 (ALDH2) genotype and male probable alcohol use disorders (AUD) was performed in Khon Kaen province, northeast Thailand. One hundred and twenty-four of cases (probable AUD) were obtained from male villagers aged 18-65 years using the modified Michigan Alcoholism Screening Test-Thai version. The same number of controls were selected, being matched with the cases in terms of age (+/-4 years) within the same village. Marital status, education history and past or present histories of physical illnesses were essentially the same for the cases and the controls. All of the cases and 85.5% of the controls were current drinkers, and the cases tended to drink significantly more often than the controls. Genomic DNA was extracted from fingernails and ALDH2 genotypes were determined by polymerase chain reaction technique and digested by Ksp 632I. The ALDH2 genotypes of the cases and the controls were not significantly different: 90.3% versus 91.1% normal homozygote; 8.1% versus 8.9% heterozygote; and 1.6% versus 0.0% mutant homozygote, respectively. Among the normal homozygote, the daily amount of alcohol intakes of the cases were significantly larger than that of the controls (56.2+/-40.6g vs. 8.1+/-14.1 g), the same was found among the ALDH2 deficient (55.9+/-43.4 vs. 2.2+/-5.8 g). Multivariate analysis based on the conditional logistic regression model showed no significant association of AUD with ALDH2 genotype, marital status, education history, or past history of injury, however, occupation and daily amount of alcohol intake were found to be significantly associated with AUD (OR = 10.72, 95% CI = 1.15-99.99, P = 0.037, and OR = 1.12, 95% CI = 1.06-1.18, P = 0.000, respectively). Non-farmers showed 10.7 times larger risk of developing AUD compared to farmers, and the subjects had three times more chance of developing AUD for each increase of 10 g of the daily amount of alcohol intake. PMID- 10459743 TI - Depression among caregivers of the disabled elderly in southern Japan. AB - The present study was conducted in an attempt to investigate factors related to depression of caregivers who looked after the disabled elderly residing in a community in southern Japan. A questionnaire survey was carried out of 45 pairs of caregivers and the disabled elderly who received regular visits from a practice nurse, who was located in Onga County, Fukuoka, in southern Japan. Twenty-four out of 45 caregivers (53.3%) scored above the conventional cut-off of the Center for Epidemiologic Studies Depression Scale (CES-D). The proportion of children-in-law caregivers was significantly smaller (0% vs. 23.8%, P = 0.01) and the proportion of caregivers who had consulted a physician within 1 month prior to the study was greater in the depressed than that in the non-depressed (79.2% vs. 47.6%, P = 0.03). The elderly whose caregivers were depressed (group 1) had more behavioural disturbances than those of the non-depressed caregivers (group 2) (1.92+/-2.38 vs. 0.52+/-0.87, P = 0.01) A multiple logistic regression analysis revealed that only behavioural disturbance was positively related to depression (Odds ratio: 4.29, 95% confidence interval: 1.28-14.37). More behavioural disturbances led the caregivers to depression or the depressed caregivers failed to take care of the elderly, which resulted in an increase in the number of the behavioural disturbances of the elderly. PMID- 10459744 TI - Neuropsychological measures in women with obsessive-compulsive disorder and trichotillomania. AB - Women with obsessive-compulsive disorder (OCD), trichotillomania (TTM) and normal controls completed a neuropsychological battery which was designed to test executive and visuospatial dysfunctions. Differences between the combined patient group and normal controls on the Rey-Osterreith Copy Test were consistent with previous work demonstrating similar neuropsychological dysfunctions in OCD and TTM. PMID- 10459745 TI - A case of seizures 1 week after the cessation of interferon-alpha therapy. AB - We report on a 60-year-old woman with a history of bipolar mood disorder who had seizures and developed a delirious state 1 week after the cessation of interferon alpha (IFN-alpha) for chronic hepatitis C. The IFN-alpha was administered to the patient for 7 weeks (266 million IU). One week after the cessation of IFN-alpha therapy, the patient had four generalized tonic-clonic seizures over a 2-day period and developed a delirious state for 2 months. We consider these seizures and delirious state to be related to IFN-alpha. PMID- 10459746 TI - A search for a mutation in the tumour necrosis factor-alpha gene in narcolepsy. AB - The discovery of almost 100% association of narcolepsy with human leukocyte antigens (HLA) DR2 antigen prompted molecular biological research of this disorder. In the HLA class II gene cluster, the gene for tumour necrosis factor alpha (TNF-alpha), which plays a role in the regulation of normal human sleep, is located. The present study searched for a mutation in the TNF-alpha gene by single-strand conformation polymorphism analysis (SSCP) in patients with narcolepsy. No mutation was detected in exons and introns of the TNF-alpha gene by SSCP and sequencing. PMID- 10459748 TI - The Montgomery Asberg Depression Rating Scale in bipolar II and unipolar out patients: a 405-patient case study. AB - The aim of the present study was to find if the Montgomery Asberg Depression Rating Scale (MADRS) can identify symptom differences between bipolar II and unipolar depression. Four hundred and five consecutive bipolar II and unipolar depressed out-patients were interviewed with the Comprehensive Assessment of Symptoms and History structured interview, following DSM-IV criteria, the MADRS, and the Global Assessment of Functioning Scale. The Montgomery Asberg Depression Rating Scale items were not significantly different between bipolar II and unipolar patients. Comparisons among atypical and non-atypical bipolar II and unipolar patients showed that only MADRS items of 'reduced sleep' and 'reduced appetite' were significantly different between atypical and non-atypical patients. PMID- 10459747 TI - Adult-type metachromatic leukodystrophy with a compound heterozygote mutation showing character change and dementia. AB - A 26-year-old Japanese woman slowly developed a change of character such as hypospontaneity and blunted affect, followed by obvious mental deterioration. She was diagnosed as having a disorganized type of schizophrenia at the first examination. Brain magnetic resonance imaging demonstrated diffuse high intensity in the cerebral white matter, particularly in the frontal lobes. The single photon emission computed tomography images using 123I-IMP disclosed diffuse cerebral hypofusion, especially in the frontal lobes. Electroencephalogram showed a moderate amount of 5-6Hz theta waves on the background of alpha activity. Nerve conduction velocities in the extremities were delayed. The level of leucocyte arylsulphatase was low. In the arylsulphatase A gene analysis, a compound heterozygote having the 99Gly-->Asp and 409Thr-->Ile mutations was confirmed. The patient was diagnosed as having metachromatic leukodystrophy. She gradually showed obvious dementing symptoms such as memory disturbance and disorientation. The characteristics of the psychiatric symptoms in the leukodystrophy are discussed. PMID- 10459750 TI - Writing for the Journal of Orthopaedic Research. PMID- 10459749 TI - Secondary personality change in psychiatric in-patients. AB - Secondary personality change (SPC) can be caused by various medical conditions and is rarely reported. We retrospectively investigated the prevalence, characteristics, clinical features and treatment response in psychiatric in patients with a primary diagnosis of SPC. Ten cases of SPC were diagnosed in a series of 5774 patients. Head trauma was the leading cause of SPC. Impulsivity, affective lability and aggression were commonly found in these SPC patients. After medication treatment, all SPC patients had mild to moderate improvement. Secondary personality change is rarely diagnosed in psychiatric in-patients and responds poorly to medication treatment. PMID- 10459751 TI - Coordinate upregulation of cartilage matrix synthesis in fibrin cultures supplemented with exogenous insulin-like growth factor-I. AB - The addition of insulin-like growth factor-I to cartilage cultures is known to stimulate the synthesis of proteoglycan and type-II collagen in explant and monolayer studies. The purpose of this study was to determine the effects of long term supplementation with insulin-like growth factor-I in chondrocytes cultured in fibrin discs as a preliminary investigation to in vivo application of chondrocyte/insulin-like growth factor-I/fibrin grafts to articular-cartilage repair procedures. Chondrocyte-fibrin cultures were maintained for 14 days, with insulin-like growth factor-I added at varying concentrations of 0, 10, 50, or 100 ng/ml medium. Cultures supplemented with 50 or 100 ng of growth factor/ml had increased levels of aggrecan and type-IIB procollagen mRNA, and translation to aggrecan and type-IIB collagen was confirmed by dye-binding assay of total proteoglycan, type-II collagen immunohistochemistry, and determination of collagen content by high-performance liquid chromatography. Maintenance of the chondrocyte phenotype during the 14 days of culture was confirmed by round cell morphology on routine staining, expression of type-II procollagen mRNA on in situ hybridization, evidence of production of pericellular type-II collagen on immunocytochemistry, synthesis of large-molecular-size aggrecan monomer on CL-2B column chromatography, and lack of appreciable message expression for type I or IIA collagen on Northern blot hybridization. Dose-response effects of insulin like growth factor-I on the expression of chondrocyte matrix constituents were most pronounced at 50 and 100 ng of growth factor per milliliter of medium. These data confirm that (a) culture of chondrocytes for extended periods in three dimensional cultures of fibrin maintains the chondrocyte phenotype and (b) supplementation with increasing concentrations of insulin-like growth factor-I enhances chondrocyte matrix synthesis and may provide a means to enhance chondrocyte phenotypic stability and function during transplantation grafting procedures. PMID- 10459752 TI - Enhanced repair of extensive articular defects by insulin-like growth factor-I laden fibrin composites. AB - Stem cells indigenous to the cancellous spaces of the bone bed in an acute injury provide an important source of pluripotent cells for cartilage repair. Insulin like growth factor-I facilitates chondrogenesis of bone marrow-derived stem cells in long-term culture and may enhance chondrogenesis in healing cartilage lesions in vivo. This study examined the impact of insulin-like growth factor-I, gradually released from fibrin clots polymerized in situ, on the recruitable stem cell pool in a full-thickness critical cartilage defect model. Twelve full thickness 15-mm cartilage lesions in the femoropatellar articulations of six young mature horses were repaired by an injection of autogenous fibrin containing 25 microg of human recombinant insulin-like growth factor-I or, in control joints, fibrin without the growth factor. All horses were killed at 6 months, and cartilage repair tissue and surrounding cartilage were assessed by histology, histochemistry, types I and II collagen immunohistochemistry, types I and II collagen in situ hybridization, and matrix biochemical determinations. White tissue filled grafted and control lesions, with the growth factor-treated defects being more completely filled and securely attached to the subchondral bone. A moderately improved chondrocyte population, more columnar cellular organization, and better attachment to the underlying bone were evident on histological evaluation of growth factor-treated defects. Type-II procollagen mRNA was abundantly present in the deeper half of the treated sections compared with moderate message expression in control tissues. Immunolocalization of type-II collagen showed a preponderance of the collagen in growth factor-treated lesions, confirming translation of type-II message to protein. Composite histologic healing scores for treated defects were significantly improved over those for control defects. DNA content in the cartilage defects was similar in treated and control joints. Matrix proteoglycan content was similar in treated and control defects and lower in the defects than in the intact surrounding and remote cartilage of the treated and control joints. The proportion of type-II collagen significantly increased in growth factor-treated tissues. Fibrin polymers laden with insulin-like growth factor-I improved the histologic appearance and the proportion of type-II collagen in healing, full-thickness cartilage lesions. However, none of the biochemical or morphologic features were consistent with those of normal articular cartilage. PMID- 10459753 TI - Low-intensity ultrasound stimulates proteoglycan synthesis in rat chondrocytes by increasing aggrecan gene expression. AB - We evaluated the effect of low intensity-pulsed ultrasound stimulation on rat chondrocytes in vitro using two different 1.0-MHz ultrasound signals with spatial and temporal average intensities of 50 or 120 mW/cm2. The pulses had a duration of 200 microseconds and were repeated every millisecond, with corresponding average peak-pressure amplitudes of 230 or 360 kPa, respectively. Cells were stimulated one, three, or five times for 10 minutes each day starting the third day after plating. One group of cells was exposed to sham ultrasound as a control. The cultures were evaluated for cell proliferation (by [3H]thymidine incorporation and DNA measurement), steady-state mRNA levels of alpha1(I) and alpha1(II) procollagens and aggrecan (by Northern blotting), and proteoglycan synthesis (by [35S]sulfate incorporation). The results revealed that ultrasound causes increases in the level of aggrecan mRNA (p < 0.05) and in proteoglycan synthesis (p < 0.03) after three and five treatments. Expression of mRNA for alpha1(II) procollagen increased over time, but ultrasound had no stimulatory effect. Expression of mRNA for alpha1(I) procollagen was initially low and remained unchanged with time. Although cell proliferation increased with time in both groups, there was no statistically significant difference between the cultures treated with ultrasound and the controls (p = 0.1). The in vitro results support our previous in vivo findings that low-intensity ultrasound stimulates aggrecan mRNA expression and proteoglycan synthesis by chondrocytes, which may explain the role of ultrasound in advancing endochondral ossification, increasing the mechanical strength of fractures, and facilitating fracture repair. PMID- 10459754 TI - Immunohistochemical analysis of type-X-collagen expression in osteoarthritis of the hip joint. AB - Conflicting data have been reported on the spatial distribution of type X collagen expression in osteoarthritis, and no concise data exist on a possible correlation between type X-collagen expression and clinical and radiological alterations. Well defined clinical and radiological data were compared with histopathological and immunohistochemical findings to investigate the expression of type-X collagen in osteoarthritis of the hip joint. Femoral heads were obtained in toto from 11 patients undergoing routine hip arthroplasty for femoral neck fractures (n = 3) or osteoarthritis (n = 8) and from 13 patients (age: 12 days to 69 years) without any evidence of hip-joint pathology. Whole coronal sections from the femoral head were decalcified for routine histology and immunohistochemical analysis with use of type-specific monoclonal antibodies to type-X collagen. Our results demonstrate that type-X collagen is consistently found in osteoarthritic cartilage and is absent from normal adult cartilage (including the region of calcified cartilage). Except for the occurrence of type X collagen in the middle zone of articular cartilage in advanced stages of osteoarthritis, there is no specific change in the staining pattern or intensity for the collagen during osteoarthritis, particularly when the staining is related to clinical and radiological parameters. Hardly more than 20% of the extracellular matrix stained for type-X collagen; therefore, we suggest that, in most cases, this type of collagen may not play a direct biomechanical role in the weakening of osteoarthritic cartilage but rather may contribute indirectly to a disturbance of the disc biomechanics by altering matrix-molecule interaction. However, expression of type-X collagen may indicate a change in chondrocyte phenotype that consistently coincides with the formation of chondrocyte clusters, one of the first alterations in osteoarthritis visible on histologic examination. PMID- 10459755 TI - Tensile properties of articular cartilage are altered by meniscectomy in a canine model of osteoarthritis. AB - Loss of or damage to the meniscus alters the pattern of loading in the knee joint and frequently leads to cartilage degeneration and osteoarthritis. The mechanical properties of articular cartilage have been shown to reflect the extent of cartilage degeneration in human osteoarthritis and in experimental models of joint disease, but there is little experimental data documenting changes in cartilage mechanics following meniscectomy. We hypothesized that the tensile properties of the surface zone of articular cartilage are altered following total medial meniscectomy. Twelve mongrel dogs underwent complete resection of the medial meniscus in the right knee, and the femoral cartilage was studied 12 weeks after the operation. We performed uniaxial, tensile stress-relaxation tests to determine the equilibrium tensile modulus of surface-zone cartilage. Water and glycosaminoglycan content were also measured at site-matched locations. The tensile moduli of the cartilage decreased significantly following meniscectomy. The linear region modulus decreased by 40%, from 25.5 +/- 7.7 to 15.3 +/- 7.2 MPa. There was a weak (r = -0.45), but significant, correlation between the linear region modulus and the gross morphological grade for cartilage damage. Water and glycosaminoglycan content did not change following meniscectomy. Composition was not correlated with mechanical properties or morphological grade, suggesting that cartilage structure may play a more important role than composition in determining the mechanical properties. The observed decrease in cartilage material properties provides a quantitative measure of the loss of cartilage function following meniscectomy and reflects a pattern of change that is consistent with damage to the collagen-proteoglycan solid network. PMID- 10459756 TI - Mechanically modulated cartilage growth may regulate joint surface morphogenesis. AB - The development of normal joints depends on mechanical function in utero. Experimental studies have shown that the normal surface topography of diarthrodial joints fails to form in paralyzed embryos. We implemented a mathematical model for joint morphogenesis that explores the hypothesis that the stress distribution created in a functional joint may modulate the growth of the cartilage anlagen and lead to the development of congruent articular surfaces. We simulated the morphogenesis of a human finger joint (proximal interphalangeal joint) between days 55 and 70 of fetal life. A baseline biological growth rate was defined to account for the intrinsic biological influences on the growth of the articulating ends of the anlagen. We assumed this rate to be proportional to the chondrocyte density in the growing tissue. Cyclic hydrostatic stress caused by joint motion was assumed to modulate the baseline biological growth, with compression slowing it and tension accelerating it. Changes in the overall shape of the joint resulted from spatial differences in growth rates throughout the developing chondroepiphyses. When only baseline biological growth was included, the two epiphyses increased in size but retained convex incongruent joint surfaces. The inclusion of mechanobiological-based growth modulation in the chondroepiphyses led to one convex joint surface, which articulated with a locally concave surface. The articular surfaces became more congruent, and the anlagen exhibited an asymmetric sagittal profile similar to that observed in adult phalangeal bones. These results are consistent with the hypothesis that mechanobiological influences associated with normal function play an important role in the regulation of joint morphogenesis. PMID- 10459757 TI - Mechanical modulation of growth for the correction of vertebral wedge deformities. AB - This study tested the following hypotheses: (a) a vertebral wedge deformity created by chronic static asymmetrical loading will be corrected by reversal of the load asymmetry; (b) a vertebral wedge deformity created by chronic static asymmetrical loading will remain if the load is simply removed; and (c) vertebral longitudinal growth rates, altered by chronic static loading, will return to normal after removal of the load. An external fixator was used to impose an angular deformity (Cobb angle of 30 degrees) and an axial compression force (60% body weight) on the ninth caudal (apical) vertebra in two groups of 12 5-week-old Sprague-Dawley rats. This asymmetrical loading was applied to all rats for 4 weeks to create an initial wedge deformity in the apical vertebra. The rats from group I (load reversal) then underwent 1 week of distraction loading followed by 4 weeks of asymmetrical compressive loading with the imposed 30 degree Cobb angle reversed. The rats from group II (load removal) had the apparatus removed and were followed for 5 weeks with no external loading. Weekly radiographs were obtained and serial fluorochrome labels were administered to follow vertebral wedging. After the initial 4-week loading period, the combined average wedge deformity that developed in the apical vertebra of the animals in both groups was 10.7 +/- 4.4 degrees. The group that underwent load reversal showed significant correction of the deformity with the wedging of the apical vertebra decreasing to, on average, 0.1 +/- 1.4 degrees during the 4 weeks of load reversal. Wedging of the apical vertebra in the group that underwent load removal significantly decreased to 7.3 +/- 3.9 degrees during the first week after removal of the load, but no significant changes in wedging occurred after that week. This indicated a return to a normal growth pattern following the removal of the asymmetrically applied loading. The longitudinal growth rate of the apical vertebra also returned to normal following removal of the load. Vertebrae maintained under a load of 60% body weight grew at a rate that was 59.4 +/- 17.0% lower than that of the control vertebrae, whereas after vertebrae were unloaded their growth averaged 102.4 +/- 31.8%. These findings show that a vertebral wedge deformity can be corrected by reversing the load used to create it and that vertebral growth is not permanently affected by applied loading. PMID- 10459758 TI - Reduced bone stress as predicted by composite beam theory correlates with cortical bone loss following cemented total hip arthroplasty. AB - Clinical and experimental evidence suggest that periprosthetic bone loss following total hip arthroplasty is caused in part by stress-shielding. Changes in bone stress in the proximal femur following implantation can be estimated with use of composite beam theory. We hypothesized that the degree of stress-shielding predicted by beam theory correlates with the magnitude of bone loss following cemented total hip arthroplasty. We analyzed cross sections from the proximal femur of 13 patients who had undergone unilateral cemented total hip arthroplasty. A matching implant was inserted contralaterally, and the cross sectional properties of the implant and bone and the bone density were determined. Bone loss was calculated on the basis of differences between contralateral (control) and ipsilateral (remodeled) sections and correlated to several beam-theory parameters calculated from the control sections: implant rigidity, bone rigidity, ratio of implant to bone rigidity, and predicted decrease in bone stress. All parameters except implant rigidity were significantly correlated with bone loss (p < 0.05). Parameters that included implant and bone properties were more strongly correlated with bone loss than were those based on bone properties alone. The predicted decrease in bone stress explained 50-60% of the variance in bone loss. The data also indicated that patients were not likely to lose substantial amounts of bone unless the reduction in bone stress exceeded a threshold value. Although limited by a small and heterogeneous sample, these results indicate that beam-theory predictions correlate with the degree of femoral resorption and should be investigated further as a means to identify patients at high risk for bone loss. PMID- 10459759 TI - Quantitative determination of joint incongruity and pressure distribution during simulated gait and cartilage thickness in the human hip joint. AB - The objective of this study was to provide quantitative data on hip-joint incongruity and pressure during a simulated walking cycle and on articular cartilage thickness in the same set of specimens. Using a casting technique in eight specimens of the human hip (age: 18-75 years), we determined the width of the joint space (incongruity) required at minimal load for contact at four phases of the gait cycle. The pressure distribution, measured with pressure-sensitive film, was determined at physiologic load magnitudes on the basis of in vivo measurements of hip-joint forces. Cartilage thickness was assessed with A-mode ultrasound. At minimal loading, the average maximum width of the joint space ranged from 1.1 to 1.5 mm in the acetabular roof, with the contact areas located ventro-superiorly and dorso-inferiorly throughout the gait cycle. At physiological loading, the width decreased and the contact areas covered the complete articular surface during midstance and heel-off but not during heel strike or toe-off. The pressure distribution was inhomogeneous during all phases, with average maximum pressures of 7.7 +/- 1.95 MPa at midstance. The cartilage thickness varied considerably throughout the joint surfaces; maxima greater than 3 mm were found ventro-superiorly. These data can be used to generate and validate computer models to determine the load-sharing between the interstitial fluid and the solid proteoglycan-collagen matrix of articular cartilage, the latter being relevant for the initiation of mechanically induced cartilage degeneration. PMID- 10459760 TI - Size and position of a single condyle allograft influence knee kinematics. AB - An optimal match for size and shape between the donor femur and the host knee is considered a critical factor influencing the outcome of a knee allograft implantation. An in vitro allograft model was developed to determine the influence of the size and position of a lateral distal femoral condylar allograft on knee kinematics. Functional knee motion was simulated in a cadaver host knee in the intact state after removing and reimplanting the native lateral condyle of the distal femur and after serially replacing the native condyle with eight donor allografts. Each allograft was first tested in an optimal position and subsequently shifted 3 mm proximal and 3 mm distal to the joint line to quantify changes in joint kinematics due to the position of the allograft. The intact knee and the knee with the ideally implanted native allograft followed similar kinematic trends. Decreasing the width of the allograft increased the valgus knee orientation at full flexion, translated the tibia posteriorly at full extension, and externally rotated the tibia throughout knee flexion. The proximal shift in allograft position increased the valgus orientation at full extension, translated the tibia posteriorly at mid-flexion, and externally rotated the tibia throughout flexion. The distal shift in position had the opposite effect on the kinematics of the proximal shift. These results indicate that improving techniques for preoperative size-matching and intraoperative allograft placement may help to reduce biomechanical complications following implantation of the allograft. PMID- 10459761 TI - Effects of isokinetic muscle activity on pressure in the supraspinatus muscle and shoulder torque. AB - Work-related shoulder pain is an increasing problem. Work in overhead positions, which causes high pressure in the supraspinatus muscle, has been shown to increase the shoulder load. It is not known how different types of muscle activity in the shoulder affect pressure in the muscle. The aim of this study was to investigate the difference between contraction modes. This has not been done before for shoulder muscles. The results are relevant from several aspects, both clinical and experimental. Pressure in the supraspinatus muscle and torque generation in the shoulder during isokinetic concentric and eccentric activity were measured in nine healthy human subjects. Torque and arm position were measured continuously with a computerized ergonometer. The contraction velocity was 60 degrees per second, and the range of movement was 0-120 degrees of abduction. Electromyographic measurements were recorded with a surface electrode above the supraspinatus muscle belly. Intramuscular pressure was recorded with a microcapillary infusion technique. Peak intramuscular pressure did not differ significantly between the two modes of contraction, but the peak was reached at 115 degrees (SD = 15 degrees) of abduction during concentric activity and at 6 degrees (SD = 7 degrees) of abduction during eccentric activity. The ratio between intramuscular pressure and torque was 3.0 mm Hg/Nm (0.40 kPa/Nm) during concentric activity and 2.3 mm Hg/Nm (0.31 kPa/Nm) during eccentric activity. The peak torque occurred at 44 degrees of abduction during concentric muscle activity and at 74 degrees of abduction during eccentric activity. Intramuscular pressure was higher during isometric contraction than during eccentric and concentric activity, and the torque was in between the two latter contraction modes. We conclude that the supraspinatus muscle is heavily loaded not only in high arm positions during concentric contraction but also during eccentric contraction in arm positions of 0-30 degrees of abduction. PMID- 10459762 TI - Substance P and calcitonin gene-related peptide expression at the extensor carpi radialis brevis muscle origin: implications for the etiology of tennis elbow. AB - With use of immunohistochemistry and antibodies to substance P and calcitonin gene-related peptide, nerve fibers showing substance P-like and calcitonin gene related peptide-like immunoreactivity were demonstrated at the origin of the extensor carpi radialis brevis muscle in patients with tennis elbow (n = 6) and in healthy controls (n = 6). The nerve fibers were distributed in association with a subpopulation of small blood vessels and in nerve bundles but were not distributed in the tunica media-adventitia junction of the arterioles. There were no inflammatory-cell infiltrates and few solitary mast cells. The present study gives further evidence to previous suggestions that tennis elbow is not an inflammatory process in the sense of involving inflammatory cells. Frequent mechanical involvement affects sensory innervation, and substance P and calcitonin gene-related peptide may have various important efferent effects, including microvascular leakage and local edema formation; therefore, the observations from this study constitute a morphological substrate for possible effects of substance P and calcitonin gene-related peptide at the origin of the extensor carpi radialis brevis muscle. PMID- 10459763 TI - Muscle regeneration and fiber-type transformation during distraction osteogenesis. AB - The successful outcome of distraction osteogenesis depends in part on the adequate adaptation of the surrounding soft tissue. We characterized the adaptation of the tibialis anterior during distraction osteogenesis at different rates (0.7 and 1.4 mm/day) and amounts (15 and 30%) of lengthening. We documented the increased expression of neonatal and slow myosin heavy chain in the tibialis anterior of skeletally immature rabbits. There was neither expression of neonatal myosin heavy chain in the experimental soleus or in the slow muscle fibers of the tibialis anterior nor increased expression of slow myosin heavy chain in the soleus or gastrocnemius. The increased amount of neonatal myosin heavy chain was concentrated in the distal half of the muscle, whereas the increase in the number of fibers that were labeled with antibodies to slow myosin occurred to the same extent throughout the tibialis anterior. Electrophysiological methods showed that the tibialis anterior was functionally intact during and after distraction osteogenesis. We concluded that in the tibialis anterior of young, skeletally immature animals, distraction osteogenesis seems to induce a recapitulation of the developmental process without leading to functional changes. In addition, during distraction osteogenesis, a fiber-type transformation occurs similar to that observed in models of muscle overloading. PMID- 10459764 TI - Late-preconditioning protection is evident in the microcirculation of denervated skeletal muscle. AB - We investigated whether ischemic preconditioning induces microvascular protection in skeletal muscle at the late phase (after 24 hours) when the same muscles are subjected to prolonged warm global ischemia. The cremaster muscle of the male Sprague-Dawley rat underwent vascular isolation and was subjected to 4 hours of ischemia and 60 minutes of reperfusion. Early preconditioning consisted of 45 minutes of ischemia followed by 15 minutes of reperfusion before prolonged ischemia/reperfusion; late preconditioning also consisted of 45 minutes of ischemia but was done 24 hours (24-hour period of reperfusion) before the prolonged ischemia/reperfusion. Arteriole diameters and capillary perfusion were measured with use of intravital microscopy. Four groups were compared: rats that underwent early preconditioning, their controls, rats that underwent late preconditioning, and their controls. Early and late preconditioning significantly attenuated vasospasm and capillary no-reflow compared with the controls for each. Average arteriole diameter was significantly larger in the rats that underwent late preconditioning than in any other rats; it was also significantly larger in the controls for late preconditioning than in those for early preconditioning. We introduce a model of the rat cremaster muscle that has been isolated from its vascular supply as a useful preparation to study the effects of late preconditioning on microcirculation in skeletal muscle. Late preconditioning provided better microvascular protection than did early preconditioning. The mechanism for this preconditioning protection is being investigated because it should provide a means for therapeutic intervention. PMID- 10459765 TI - Dynamic measurement of bone blood perfusion with modified laser Doppler imaging. AB - Although the mechanisms are not clearly defined, blood flow may play an important role in moderating skeletal adaptation. Most techniques currently available to measure blood flow in bone are time-consuming and require destruction of the tissue, but laser Doppler technology offers a less invasive method. The present study assessed whether laser Doppler perfusion imaging could detect changes in perfusion in cortical bone. By use of modified laser Doppler perfusion imaging with an adjustable, incorporated, near infrared-laser gain photodetection system, perfusion of blood in the mid-diaphyseal tibial cortex of New Zealand White rabbits (n = 5) was measured before, during, and after occlusion of the femoral artery. During occlusion, perfusion decreased 69% compared with control levels; removal of the arterial clip caused flux values to return to near normal. Laser Doppler perfusion imaging provides a two-dimensional image related to blood flow, and the results of this pilot study suggest that it may be an effective technique for imaging in vivo dynamic changes in perfusion in cortical bone. PMID- 10459766 TI - Uniaxial yield strains for bovine trabecular bone are isotropic and asymmetric. AB - Although evidence suggests that yield strains for trabecular bone are isotropic, i.e., independent of loading direction, decisive support for this hypothesis has remained elusive. To explicitly test whether yield strains for trabecular bone are isotropic, compressive and tensile yield strains of 51 specimens of bovine tibial trabecular bone (0.41 +/- 0.08 g/cm3 [mean apparent density +/- SD]) were measured without end artifacts in on-axis (along the principal trabecular orientation) and off-axis (30-40 degrees oblique to on-axis) orientations. Yield strains for the on-axis and off-axis orientations were similar in tension (0.80 +/- 0.03% compared with 0.85 +/- 0.04%, p = 0.21) and compression (0.97 +/- 0.05% compared with 0.96 +/- 0.07%, p > 0.99); as expected, modulus and strength depended on loading direction. When considered with an ancillary experiment on bovine tibial trabecular bone that showed yield strains to be similar between on axis and 90 degrees off-axis bone, these results firmly establish the isotropy of uniaxial yield strains for bovine tibial trabecular bone. This bone is of high density and has a strong, plate-type, anisotropic architecture. Therefore, yield strains for uniaxial loading are expected to be isotropic, or nearly so, for other types of dense trabecular bone, although further work is required to confirm this and to establish this behavior for bone of lower density. PMID- 10459767 TI - Effects of sintered bovine bone on cell proliferation, collagen synthesis, and osteoblastic expression in MC3T3-E1 osteoblast-like cells. AB - To determine the function of biomaterials on osteoblasts, we studied the effects of two different biomaterials, sintered bovine bone (true bone ceramic) and hydroxyapatite-related material, on DNA and collagen synthesis and osteoblastic expression in osteoblast-like cells in vitro. Osteoblasts cultured with true bone ceramic exhibited greater increases in DNA and collagenous protein synthesis and alkaline phosphatase activity than those cultured with hydroxyapatite. Furthermore, expression of mRNA for type-I collagen and osteocalcin, as assessed by reverse transcription-polymerase chain reaction, was higher in cultures grown on true bone ceramic or hydroxyapatite than in those grown with glass fragments, and levels of gene expression in true bone ceramic and hydroxyapatite cultures were at almost the same level. These findings indicate that osteoblasts cultured with true bone ceramic increase their activity, suggesting that true bone ceramic may be a more favorable substrate than hydroxyapatite for growth and differentiation of osteoblast-like cells. PMID- 10459768 TI - Pertussis toxin-sensitive G proteins as mediators of stretch-induced decrease in nitric-oxide release of osteoblast-like cells. AB - Mechanical loading plays an important role in regulating bone remodeling, and nitric oxide may be one regulator of this process. To determine how mechanical stress modulates osteoblast function, we loaded cyclic tensile stretch on osteoblast-like cells and measured levels of nitric oxide in the medium. High frequency of stretch at any magnitude inhibited release of nitric oxide; however, low frequency of stretch enhanced its release from the static control. To examine the involvement of G protein (guanine nucleotide-binding regulatory protein) in stress-inhibited release of nitric oxide, we added pertussis toxin, a specific inhibitor of the Gi class, and found that it completely reversed the stress inhibited release. These data support the idea that pertussis toxin-sensitive G protein is activated in the presence of cyclic tensile stretch. PMID- 10459769 TI - Effect of demineralized bone matrix on polymorphonuclear leukocyte degranulation. AB - The potential use of allogenic demineralized bone matrix to augment or treat bone defects or nonunions in animals and humans is currently being investigated. Demineralized bone matrix induces osteogenesis by a multistep cascade of endochondral ossification that is mediated by bone-induction factors. The migration and activation of polymorphonuclear leukocytes appear to be critical in the initiation of the cascade of osteogenesis induced by demineralized bone matrix. This study examined the effects of demineralized bone matrix on the degranulation of polymorphonuclear leukocytes. Demineralized bone matrix stimulated the release of polymorphonuclear leukocyte-specific, but not azurophilic, granules in a time and dose-dependent manner. The ability of the bone matrix to induce this degranulation was independent of its size and species. The mechanism by which this degranulation occurs is not completely understood; however, it is known that it does not occur by means of a receptor that requires guanidine triphosphate-dependent regulatory proteins as does polymorphonuclear leukocyte degranulation induced by N-formyl peptide. The factor that stimulates degranulation is not type-I collagen but rather appears to be a cytokine that has a heparin-binding domain and a molar mass of 10-70 kDa. Loss of the ability of demineralized bone matrix to induce degranulation of polymorphonuclear leukocytes correlated positively with the loss of its ability to induce bone formation. PMID- 10459770 TI - Novel formulation of fibroblast growth factor-2 in a hyaluronan gel accelerates fracture healing in nonhuman primates. AB - Recent advances in understanding the biology of fracture healing and the availability of specific macromolecules has resulted in the development of novel treatments for injuries to bone. Fibroblast growth factor-2 or basic fibroblast growth factor (4 mg/ml), a potent mitogen, and hyaluronan (20 mg/ml), an extracellular matrix component, were combined into a viscous gel formulation intended for direct, percutaneous injection into fresh fractures. In an experimental primate fracture model, a bilateral 1-mm-gap osteotomy was surgically created in the fibulae of baboons. A single direct administration of this hyaluronan/fibroblast growth factor-2 formulation to the defect site significantly promoted local fracture healing as evidenced by increased callus formation and mechanical strength. Radiographic analysis showed that the callus area was statistically significantly larger at the treated sites than at the untreated sites. Specimens treated with 0.1, 0.25, and 0.75 ml hyaluronan/fibroblast growth factor-2 demonstrated a 48, 50, and 34% greater average load at failure and an 82, 104, and 66% greater energy to failure than the untreated controls, respectively. By histologic analysis, the callus size, periosteal reaction, vascularity, and cellularity were consistently more pronounced in the treated osteotomies than in the untreated controls. These results suggest that hyaluronan/fibroblast growth factor-2 may provide a significant advance in the treatment of fractures. PMID- 10459771 TI - Integrin subunits responsible for adhesion of human osteoblast-like cells to biomimetic peptide surfaces. AB - We have identified the integrin subunits responsible for the initial adhesion of human osteoblast-like cells to peptide-modified surfaces. Biomimetic peptide surfaces containing homogenous RGD (Arg-Gly-Asp), homogenous FHRRIKA (Phe-His-Arg Arg-Ile-Lys-Ala), and a mixed ratio of FHRRIKA:RGD (25:75) were used to assess integrin-mediated adhesion. The RGD and FHRRIKA peptides were selected from the cell-binding and putative heparin-binding domains of bone sialoprotein. A panel of monoclonal antibodies against human alpha1, alpha2, alpha3, alpha4, alpha5, beta1, alpha(v), and alpha(v)beta3 was used to identify the subunits most dominant in mediating short-term (10 or 30 minutes) and long-term (4 hours) cell adhesion to the peptide surfaces. Anti-alpha2, anti-beta1, and anti-alpha(v) significantly (p < 0.05) diminished cell attachment to homogenous RGD surfaces following 30 minutes of incubation. After 4 hours of incubation on RGD-grafted surfaces, immunostaining of these integrin subunits revealed discrete localization of the alpha(v) subunit at the periphery of the cell (similar to focal contact points), whereas the alpha2 and beta1 subunits stained very diffusely throughout the cell. A radial-flow apparatus was used to determine the effect of anti-integrin antibodies on strength of cell detachment following 10 minutes of incubation on peptide-grafted surfaces. The strength of detachment from surfaces containing RGD was significantly reduced (p < 0.05) in the presence of anti-alpha2, anti-alpha(v), or anti-beta1 compared with controls (presence of preimmune mouse IgG). None of the antibodies significantly influenced cell attachment to homogenous FHRRIKA-grafted surfaces. These results demonstrate that initial (30 minutes) attachment of human osteoblast-like cells to homogenous RGD surfaces was mediated by the collagen receptor alpha2beta1 and the vitronectin receptor alpha(v)beta3, whereas only the vitronectin receptor governed longer term (longer than 30 minutes) adhesion (localization to focal contacts). The importance of distinct integrins in mediating the attachment of bone cells to RGD immobilized surfaces indicates a strategy for engineering orthopaedic implants with a built-in surface specificity for cell adhesion. PMID- 10459772 TI - Elastic fibers of musculoskeletal tissues in bovine Marfan syndrome: a morphometric study. AB - Bovine Marfan syndrome has all the major pathognomonic, clinical, and pathological features of the human syndrome. To further explore the alterations characterizing Marfan syndrome, musculoskeletal tissues from affected and healthy cattle were subjected to histochemical and histomorphometric analysis. Our findings demonstrated reduced elastic fiber content in the periosteum, capsule, interosseous ligament, and flexor tendons of the metatarsophalangeal joint of affected cattle. The elastic properties of a tissue are supplied by elastic fibers in the extracellular matrix; therefore, their reduced content in articular tissues suggests that these fibers could be functionally incompetent to withstand normal stress, causing a predisposition to joint laxity and dislocation. Moreover, perichondrial-periosteal membranes, which are made of collagen and elastic fibers, are known to affect the growth process of the long bones. The decrease in the number of elastic fibers in these membranes could result in reduced restraint of skeletal growth and explain some skeletal abnormalities of Marfan syndrome (i.e., dolichostenomelic habitus). PMID- 10459773 TI - Deposition and retention of vital and dead Streptococcus sanguinis cells on glass surfaces in a flow-chamber system. AB - The proportion of vital as compared with dead Streptococcus sanguinis cells attached to glass surfaces was monitored and related to varying proportions of planktonic vital as compared with dead Strep. sanguinis cells. In a flow chamber with six parallel-mounted glass plates, Strep. sanguinis was suspended in pretreated sterile human saliva. Deposition of Strep. sanguinis took place, with a proportion of vital sanguinis streptococci in saliva (%VSs) of 90%, 45% or 22.5%. After exposure times of 30, 60, 90, 120 and 240 min, adherent microorganisms were labelled with two fluorescence stains to differentiate between vital and dead bacteria. Proportions of vital attached streptococci (%VSa) were determined microscopically. Dead bacteria were detected on all glass plates. The %VSa at 30 min and 60 min was significantly lower than the baseline %VSs. During the course of a single run the %VSa frequently increased after either 30, 60 or 90 min without exceeding the %VSs at 4 h. %VSs was the only variable exerting a significant effect on %VSa at 30 and 60 min. It is suggested that during the initial events of microbial attachment the dead rather than vital Strep. sanguinis cells attach preferably to solid surfaces. PMID- 10459774 TI - Formation and mineralization of murine molar roots with hypocalcaemia induced by a low-calcium diet and the changes after returning to a normal diet. AB - The purpose was to examine the formation and mineralization of molar roots in rats with hypocalcaemia, induced by a low-calcium diet and the changes after returning to a normal diet. Two-week-old male Wistar rats were fed a low-calcium diet (0.03% calcium) during the period when the first molar roots form and then a diet containing normal amounts of calcium (1.83% calcium) was restored. The blood calcium, the length and mineralization of the first upper molar roots, and the density of surrounding alveolar bone were measured, and the morphological and histological features of the roots examined. While the rats were fed the low calcium diet, they had a significantly lower blood calcium than normal controls. Morphologically, the upper first molar roots were shorter and the predentine layer was thicker. The mineralization of dentine and the surrounding alveolar bone was significantly less than in the controls and no mineralization was detected in the thickened predentine. After a normal calcium diet had been restored, the blood calcium, thickness of dentine, alveolar bone density, and length of the roots caught up with the normal. In addition, hypomineralized dentine and interglobular dentine were observed. Subsequently, the mineralization of the dentine increased and the amount of interglobular dentine gradually decreased. These results suggest that in rats a low-calcium diet induces hypocalcaemia, causing the formation of interglobular dentine and hypomineralization of the dentine of the roots. Most physiological variables recovered completely with the return to a normal-calcium diet, although some hypomineralization of the dentine remained. PMID- 10459775 TI - Haemodynamic changes in human masseter and temporalis muscles induced by different levels of isometric contraction. AB - This study evaluated the influence of low contraction forces on intramuscular haemodynamics in human masseter and temporalis using near-infrared tissue spectroscopy. This method allowed the intramuscular haemoglobin (Hb) to be assessed dynamically before, during and after a 5, 15, 25 and 100% maximum voluntary contraction (MVC). Twenty volunteers, 10 males and 10 females, without pain or dysfunction in the masticatory system were included in this study. Data were recorded for 30 s before, 30 s during and 5 min after the four sustained contraction tasks. The results showed that all four levels of voluntary contraction produced a clear haemodynamic response (during and after contraction) in both muscles. For analytical purposes, the maximum Hb achieved after 100% MVC was set equal to 1.00. In the masseter the mean peak Hb during the 5, 15, 25 and 100% MVC was 0.49, 0.92, 1.30 and 1.73 while after the contractions it was 0.50, 0.65, 0.78 and 1.00, respectively. In the temporalis the peak Hb during the contractions was 0.23, 0.36, 0.48 and 0.66 and after the contractions 0.32, 0.45, 0.56 and 1.00, respectively. Repeated-measures analysis of variance revealed a significant main effect for the different contraction levels both in the masseter (during contraction, p = 0.001; after contraction, p<0.001) and the temporalis (during contraction, p = 0.002; after contraction, p<0.001). These data suggest that low levels of contraction induce a clear haemodynamic response, even at 5% effort. When compared, the masseter and anterior temporalis showed clearly different patterns for the Hb signal during the contraction (p<0.001) as well as after it (p = 0.007). Specifically, the Hb during the contractions in the masseter appeared more stable than in the temporalis, which showed a strong return to baseline. Obviously the contracting masseter had a stronger and more sustained venous occlusion than the contracting temporalis. It is speculated that variation in architecture between the two muscles contributes to these differences in blood flow. PMID- 10459776 TI - Effect of 5-hydroxytryptamine-2 and alpha-adrenergic receptor antagonists on the 5-hydroxytryptamine-induced decrease in rabbit masseter muscle blood flow. AB - A previous study showed a significant decrease in blood flow in the rabbit masseter during infusion of 5-hydroxytryptamine (5-HT) (10(-9) mol/l). The aim of the present study was to test the hypothesis that blockade of the 5-HT2 or the alpha-adrenergic receptor would inhibit the 5-HT-induced decrease of microcirculatory blood flow in the masseter. In 12 rabbits, the masseters were infused with 5-HT (10(-9) mol/l) in combination with the alpha-adrenoceptor antagonist phentolamine (10(-6) mol/l) or the 5-HT2-receptor antagonist ritanserin (10(-6) mol/l). The effect on microcirculatory blood flow was measured by laser-Doppler flowmetry. Infusion of 5-HT induced a significant decrease in blood flow. Inclusion of ritanserin in the 5-HT infusion solution significantly inhibited this decrease, while inclusion of phentolamine did not. This study therefore showed that the 5-HT2 receptor antagonist ritanserin inhibited the 5-HT induced decrease in microcirculatory blood flow in the rabbit masseter. This decrease in blood flow is thus mediated by the 5-HT2 receptor. PMID- 10459777 TI - Mandibular posture during sleep in patients with obstructive sleep apnoea. AB - Vertical mandibular posture is thought to be related to narrowing of the upper airway, because mouth opening is associated with an inferior-posterior movement of the mandible and the tongue which influences pharyngeal airway patency. To test whether the mandibular posture is related to the occurrence and/or termination of obstructive sleep apnoea (OSA), the vertical mandibular position was recorded intraorally using a magnet sensor during a standard sleep study in seven patients with OSA. Measurements were recorded during sleep both in the supine and lateral recumbent positions. The percentage of total sleep time spent with mandibular opening greater than 5 mm was significantly larger (p<0.001) in patients with OSA (69.3+/-23.3%) compared with our previous results obtained from healthy adults without OSA (11.1+/-11.6%). The stage of sleep affected the vertical mandibular posture during sleep in the supine position, but not in the lateral recumbent position in patients with OSA. In non-rapid eye-movement sleep, mandibular opening increased progressively during apnoeic episodes and decreased at the termination of apnoeic episodes. In contrast, no significant change in mandibular posture occurred in apnoeic episodes during rapid eye-movement sleep. It was concluded that the vertical mandibular posture is more open during sleep in patients with OSA than in healthy adults and that mandibular opening increases progressively during apnoeic episodes and decreases at the termination of those episodes. PMID- 10459778 TI - Negative correlation between oral malodour and numbers and activities of sulphate reducing bacteria in the human mouth. AB - The majority of cases of oral malodour are thought to be due to bacterial activities in the mouth, but many of the bacterial species responsible have not been identified. Volatile sulphide compounds have been proposed as constituents of oral malodour. Therefore, the relation between intensity of odour and numbers of bacteria in the mouth that are sulphide-producing from sulphate was investigated. Numbers of such dissimilatory sulphate-reducing bacteria (SRB) and sulphide reduction rates were evaluated in samples from different oral sites in relation to measures of oral malodour. Results showed that sulphate-reducing bacterial numbers and activities were negatively correlated with malodour, as determined by organoleptic assessment and measurement with a sulphide-monitoring instrument, the Halimeter. The data indicate that sulphide produced by oral SRB may not be an important contributor to oral malodour. A rather poor correlation was observed between Halimetric and organoleptic values, indicating that these methods may measure different aspects of oral malodour intensity. PMID- 10459779 TI - Simultaneous recording of mandibular condylar movement and single motor-unit activity at verified sites in the human lateral pterygoid muscle. AB - In recent years, understanding of normal jaw-muscle function has been enhanced by detailed descriptions of their complex internal architecture and of the functional activity of single motor units (SMUs). The lateral pterygoid muscle, however, has been poorly studied, although it is thought to play an important part in the control of jaw and jaw-joint movement. The present study is the first of a series of SMU studies to clarify the normal function of this muscle. The aims were to demonstrate (a) the unequivocal isolation of SMU activity from one or two verified recording sites within the lateral pterygoid, and (b) that these SMUs can be recorded reliably together with condylar movement during simple command jaw movements. Recordings of SMU activity were made with fine-wire electrodes from sites within the superior and inferior heads of the right lateral pterygoid during biting or command lateral jaw movements and combined with recordings of condylar and mid-incisor point movement. Recording sites were verified by computed tomography. In four young adults, the activities of 17 SMUs were reliably discriminated at seven recording sites within the lateral pterygoid. The units could be recorded during repeated trials of the same movement throughout a recording session with no appreciable change in amplitude or waveform. Units could also be discriminated simultaneously at separate recording sites--one in the superior head and the other in the inferior head. These data demonstrate that SMU activity can be recorded from verified sites within the lateral pterygoid simultaneously with condylar movement during command jaw movements. PMID- 10459780 TI - Substance P in the hypoglossal nucleus of the rat. AB - The distribution of substance P (SP)-containing synaptic terminals in the hypoglossal nucleus (XII) of adult rats was examined by retrograde peroxidase labelling and immunocytochemistry. From the location of peroxidase injections into the tongue and of labelled neurones in the ventral lamina of XII, motor neurones that supply intrinsic vertical, longitudinal and transverse fibres as well as the extrinsic muscle genioglossus appear to have been labelled. SP containing terminals were found making contact, and sometimes dual synapses, with unlabelled neuronal dendrites but not with retrogradely labelled somata or dendrites. These findings suggest that SP terminals may contact dendrites of interneurones or of neurones supplying other extrinsic muscles located in the anterior part of the tongue. Dual SP-containing synapses between XII motor neurones may be the means by which tongue muscle fibres are recruited and their function synchronized. PMID- 10459781 TI - Effects of cyclosporin A on the mandibular condylar cartilage in rats. AB - Twenty, 5-week-old, male, Sprague-Dawley rats were divided into a control and a cyclosporin A (CSA) group for evaluating effects of the drug on condylar cartilage. Animals in the treatment group daily received CSA (15 mg/kg body wt) in mineral oil by gastric feeding over a 4-week observation interval. Control animals received mineral oil only. Five animals from each group were killed at weeks 2 and 4 of study. After histological processing, five tissue sections from the mid-region of the condyle were selected and examined. Three compositional zones (articular fibrous, proliferative, and hypertrophic) of the superior, posteriosuperior and posterior regions of the condylar cartilage were evaluated by light microscopy. At week 2, total condylar cartilage thickness was similar in the CSA and control groups, but the thickness of each zone was altered in CSA treated animals, including a decrease of the fibrous and proliferative zones and an increase in hypertrophic zone compared to control (P<0.05). At week 4, CSA treated animals exhibited overall decreased cartilage thickness, including decreased thickness of each zone compared to control (P<0.05). The results suggest that CSA has an inhibitory effect on the maturation of the mandibular condyle in rats. PMID- 10459782 TI - Depression, cognition and quality of life in parkinsonian patients. PMID- 10459783 TI - The importance of psychological symptoms in Parkinson's disease. PMID- 10459784 TI - Ageing and the baroreflex. PMID- 10459785 TI - Measuring the impact of Parkinson's disease with the Parkinson's Disease Quality of Life questionnaire. AB - OBJECTIVES: To assess the validity of the Parkinson's Disease Quality of Life (PDQL) questionnaire, a patient-specific multi-dimensional quality of life measure, in a community-based sample of patients with Parkinson's disease (PI)) using standardized measures of disease severity, depressive symptomatology and cognitive function. DESIGN: A group of 194 patients with probable PD were randomly selected from a community-based register and were invited to self complete the 37-item PDQL. Disease severity was measured by the disease-specific Webster scale, cognition by the CAMCOG neuropsychological test and depressive symptomatology by the self-report 15-item GDS-15 geriatric depression scale. RESULTS: A total of 136 patients returned completed PDQL questionnaires. Significant differences (P < 0.05) emerged between the pooled PDQL score of patients grouped on the basis of disease severity. Depressive symptoms and cognition were also associated with poorer perceived quality of life as measured by the PDQL. CONCLUSIONS: The results of this study are indicative of the validity of the PDQL as an important additional measurement which reflects the impact of PD from the patient perspective. It shows poorer quality of life to be associated with increasing age, disease severity more severe depressive symptomatology and impaired cognitive functioning. However, the responsiveness of this instrument in the evaluation of care in PD remains to be determined. PMID- 10459786 TI - Older subjects show no age-related decrease in cardiac baroreceptor sensitivity. AB - OBJECTIVE: To examine the relationship between age, blood pressure and cardiac baroreceptor sensitivity derived from spectral analysis, the Valsalva manoeuvre and impulse response function. METHODS: We studied 70 healthy normotensive volunteers who were free from disease and not taking medication with cardiovascular or autonomic effects. We measured beat-to-beat arterial blood pressure and used standard surface electrocardiography to record pulse interval under standardized conditions with subjects resting supine as well as during three Valsalva manoeuvres. We performed single, multiple and stepwise regression of patient characteristics against cardiac baroreceptor sensitivity results. RESULTS: There is a non-linear decline in cardiac baroreceptor sensitivity with advancing age, increasing systolic blood pressure and heart rate values (except for the Valsalva-derived result), but little further decline after the fourth decade. Only age significantly influenced values derived using the Valsalva manoeuvre and impulse response analysis. Using spectral analysis, age, systolic and diastolic blood pressure and heart rate influenced cardiac baroreceptor sensitivity, age contributing to 50% of the variability. Age also influenced the relationship between pulse interval and blood pressure, possibly indicating more non-baroreceptor-mediated changes with advancing age. CONCLUSIONS: Although age is the dominant factor influencing cardiac baroreceptor sensitivity in this normotensive population, there is little change in mean values after 40 years of age. The differences in the relationship between pulse interval and blood pressure with advancing age have implications for the calculation of cardiac baroreceptor sensitivity using spectral analysis. PMID- 10459787 TI - Additional risk factors in atrial fibrillation patients not receiving warfarin. AB - OBJECTIVE: To study how many elderly inpatients with previously diagnosed atrial fibrillation were not receiving anticoagulant prophylaxis, and the prevalence of additional risk factors in these patients. METHODS: All new admissions to a department of medicine for the elderly were screened for atrial fibrillation. Additional risk factors were analysed in those with established atrial fibrillation who were not receiving warfarin. Previous hospital admissions, documentation of why prophylaxis was not being used and use of aspirin as an alternative agent were also examined. RESULTS: 56 patients had previously diagnosed atrial fibrillation; 82% were not taking warfarin and 71% of these were not on aspirin either. All patients not taking warfarin had one additional risk factor for stroke and 95% had two or more. Fifty-two percent had attended hospital when atrial fibrillation was present within the previous 3 years and there was nothing documented in their records to explain why anticoagulation had not been used. CONCLUSIONS: Most elderly inpatients with established atrial fibrillation were not taking warfarin. All had additional risk factors for stroke, which increase the absolute benefit of anticoagulation. PMID- 10459788 TI - The presentation, treatment and outcome of renal cell carcinoma in old age. AB - OBJECTIVE: To review clinical presentation and outcome of patients with a diagnosis of renal cell carcinoma at a district general hospital and assess whether older patients were more likely to present in a non-specific manner or receive more conservative management and whether their survival was less favourable. SUBJECTS AND METHODS: 39 patients presenting with a diagnosis of renal cell carcinoma between 1987 and 1995 were identified from hospital activity analysis data and histopathology records. We divided the subjects into young patients (< 69 years: n = 27) and elderly patients (>70 years: n = 10), and made a retrospective analysis of clinical features, laboratory results, pathology, staging, treatment and survival from hospital records. RESULTS: Anaemia, hypertension and weight loss were common clinical features in both young and elderly groups. The prevalence of non-urological symptoms did not differ between study groups. Anaemia was frequently microcytic and hypochromic. Hypertension was present in 46% of patients and one-third of these were newly diagnosed. In 19% of patients with renal cell carcinoma, the diagnosis was made incidentally while imaging for other indications. Elderly patients were as likely to receive surgical treatment as younger patients. Survival differed with stage but not age. CONCLUSIONS: Neither clinical presentation, management nor survival differed between the young and elderly subjects. Renal cell carcinoma should be considered in elderly patients with systemic features such as malaise or weight loss associated with anaemia, hypertension and raised erythrocyte sedimentation rate. PMID- 10459789 TI - Age is no contraindication to thyroid surgery. AB - BACKGROUND: There is increased risk of surgical treatment in elderly patients, but little has been written on the safety and efficacy of thyroid surgery in this group. OBJECTIVE: To determine if there is an increased risk of morbidity and mortality of thyroid surgery in patients over 75 years old. PATIENTS AND METHODS: During the period 1986-96, 1631 patients aged 50 and over underwent 1673 procedures by two surgeons in our unit. We retrospectively compared indications for surgery, procedures, pathology complications and mortality in three groups: 50-60-year-olds (725 patients), 61-74-year-olds (685 patients) and those aged 75 years and over (221 patients) RESULTS: The main indication for surgery in all three groups was compression or risk of malignancy. Total thyroidectomy was the main procedure performed. Although benign multinodular goitre was the commonest diagnosis, the over-75-year-olds had fewer benign multinodular goitres than the 61-74 group and more malignancy than the other two groups. There was no significant difference in mortality between the three groups. CONCLUSIONS: Surgery in patients over 75 is as safe as in younger patients with no increase in morbidity and mortality. Benign multinodular goitre is the most common indication for surgery. PMID- 10459790 TI - Lack of association between Helicobacter pylori infection and extracardiac atherosclerosis in dyspeptic elderly subjects. AB - BACKGROUND: There are conflicting data on the association between Helicobacter pylori (HP) infection and cardiovascular diseases. AIM: To determine if there is an association between gastric HP infection and atherosclerosis of cerebral or peripheral arteries in elderly subjects. METHODS: 90 dyspeptic elderly subjects had upper gastro-intestinal endoscopy and the gastroduodenal pathology was identified. HP infection was confirmed by gastric histology and the rapid urease test. Vascular ultrasonography of extracranial cerebral arteries and leg arteries was performed to evaluate (i) the presence of an atherosclerotic lesion, (ii) the total length of all plaques documented and (iii) the number of arteries with atherosclerotic lesions. Statistical analysis was by the chi2 test, Yates's corrected chi2 test, the Mann-Whitney test and logistic regression. RESULTS: 59 subjects were HP-positive. These had a higher prevalence of peptic ulcer disease (P = 0.01) and higher serum levels of IgG anti-HP antibodies (P = 0.0001), but no significant differences in the number of atherosclerotic lesions, the total length of the plaques or the number of arteries with lesions. No significant association of HP positivity was found with diabetes mellitus, hypertension, cigarette smoking or coronary heart disease, nor with serum concentrations of HDL cholesterol, fibrinogen, triglycerides or glucose. CONCLUSIONS: Elderly dyspeptic subjects with gastric HP infection had significantly more peptic ulcer disease but no more atherosclerotic lesions than those who were HP-negative. Atherosclerosis was not associated with HP infection. In this cross-sectional study of elderly patients with dyspepsia, no association between HP infection and extracardiac atherosclerosis was found. PMID- 10459792 TI - How ageing and social factors affect memory. AB - OBJECTIVES: To explore the relationships between lifestyle and memory, and determine whether social factors influence memory. METHODS: the relationship between memory and lifestyle was examined in 497 adults aged 25-80 years, using the Mectamemory in Adulthood questionnaire. We asked about sports activity and perceived activity, participation in voluntary organizations and social contacts. RESULTS: Activity and frequent contact with friends and family were related to higher memory capacity scores. Those with higher capacity scores were also younger, had better health and a stronger internal locus of control. In contrast, people with higher anxiety scores had more symptoms and less education, and were more externally oriented. CONCLUSIONS: people who consider themselves socially and physically active also consider their memory capacity to be good and are less anxious about their memory than less socially and physically active people. Perceived memory change appears to be predominantly influenced by ageing, whereas memory capacity and memory anxiety are more influenced by social factors. PMID- 10459791 TI - Declining physical abilities with age: a cross-sectional study of older twins and centenarians in Denmark. AB - OBJECTIVE: To evaluate whether physical disability reaches a plateau in the oldest age groups. DESIGN: Cross-sectional survey. PARTICIPANTS: A total of 3351 individuals, which included all those living in Denmark who celebrated their 100th anniversary during the period from 1 April 1995 to 31 May 1996 (276 subjects) and all Danish twins aged 75-94 registered in the Danish Twin Register (3075 subjects). MAIN OUTCOME MEASURES: The ability to perform selected items of basic activities of daily living independently. RESULTS: The prevalence of independence in each of six selected activities of daily living was significantly lower in both men and women centenarians compared with octo- and septuagenarians. The sex difference in independence in all six selected activities of daily living was larger for each advancing age group, with women being most disabled (P < 0.001). In centenarians 20% of women and 44% of men were able to perform all selected activities of daily living independently. CONCLUSION: Compared with individuals aged 75-79 years, physical abilities of men and women gradually diminished in age groups 80-84, 85-90 and 90-94, with the lowest levels among 100 year-olds. Although women have lower mortality, they are more disabled than men, and this difference is more marked with advancing age. PMID- 10459793 TI - Depressive illness, depressive symptomatology and regional cerebral blood flow in elderly people with sub-clinical cognitive impairment. AB - BACKGROUND: Depressive illness in dementia is often assumed to be a unitary clinical phenomenon. AIM: To describe changes in patterns of depressive symptomatology with time, and associated changes in cerebral blood flow to the frontal and temporal regions. METHOD AND RESULTS: 397 elderly people with sub clinical cognitive dysfunction were observed over 3 years. Sixteen percent of them developed dementia during the study The prevalence of depressive symptomatology was higher in this group than in the general population, especially in women, who also had higher recovery rates. A changing profile of depressive symptoms was found in depressed elderly people progressing to dementia, with fewer affective symptoms and increases in agitation and motor slowing. These changes were paralleled by greater reductions in left temporal regional cerebral blood flow than in non-depressed subjects with Alzheimer's disease. CONCLUSION: In dementia, there may be two separate and interacting depressive syndromes whose differentiation may be clinically important. PMID- 10459794 TI - Chromosomal damage and ageing: effect on micronuclei frequency in peripheral blood lymphocytes. AB - BACKGROUND: Instability in the organization and expression of the genetic material has been hypothesized as the basic mechanism of ageing. OBJECTIVE: To quantify the effect of ageing on chromosomal damage as measured by spontaneous micronuclei (MN) frequency in peripheral blood lymphocytes. METHOD: Analysis of a large population sample from two laboratories applying the cytokinesis-block technique and a third using traditional interphase analysis. The age-related effect on baseline level of micronuclei frequency and on cell proliferation measures was further investigated in a study of peripheral blood samples from healthy subjects. RESULTS: There was an increase of MN frequency with age. The regression lines showed a positive slope and were statistically significant (P< 0.01) with a steeper trend for cytochalasin B-treated samples. An inverse correlation with age was detected for the percentage of binucleated cells in laboratories using cytochalasin B. This study confirms the increase of basal level of MN with age. A decrease by age in proliferation efficiency measured by the percentage of binucleated cells suggests an interference of age-related factors on cell division. CONCLUSION: There is an increase in MN frequency with increasing age. PMID- 10459795 TI - Arnold-Chiari malformation with syringomyelia in an elderly woman. AB - PRESENTATION: A 76-year-old woman, complaining of leg pain and unsteady gait for 3 years, presented with a spastic paraparetic gait, severe spasticity and touch, thermal and pain sensory loss limited to arms, lower thorax and upper abdomen. Brain and spinal cord magnetic resonance imaging showed a large loculated syrinx. Cerebellar tonsillar herniation into the foramen magnum was also seen (Arnold Chiari malformation, type I). OUTCOME: The patient had successful cervico-spinal surgical decompression which resulted in marked reduction in hypertonia and weakness, normal gait and normal joint movement at 6 months. CONCLUSION: This unusual, late clinical presentation of a congenital disease underlines the importance of a comprehensive diagnostic work-up in the elderly patients with complex neurological signs. PMID- 10459796 TI - Extrapyramidal features in Alzheimer's disease. PMID- 10459797 TI - Indices of dehydration in elderly people. PMID- 10459798 TI - Helicobacter pylori infection in elderly patients with peptic ulcer. PMID- 10459800 TI - Are rates of ageing determined in utero? PMID- 10459799 TI - Euthanasia and old age. PMID- 10459801 TI - Outcomes of percutaneous endoscopic gastrostomy feeding. PMID- 10459802 TI - Outcomes of percutaneous endoscopic gastrostomy feeding. PMID- 10459803 TI - Pneumococcal macrolide resistance--myth or reality? AB - There is no doubt that owing to the prolific use of the macrolides and azithromycin over the past several years, resistance has developed and is increasing in incidence. I believe we should re-evaluate the use of these antibiotics for our patients and consider parameters other than the negative in vitro results. Firstly, microbiology laboratories should return to the habit of providing the clinician with MIC values for pathogenic isolates rather than generic susceptibility reports ((S)usceptible, (I)ntermediate, (R)esistant) that are based on standard disc diffusion testing. Although agar dilution MIC testing is a bulky and labour intensive practice, it provides the best data when conducted in the appropriate environment. Secondly, and more importantly, these MIC values need to be compared with in-vivo antibiotic pharmacokinetics and pharmacodynamics. Although it is possible to compare MIC values directly with serum concentrations of beta-lactams and aminoglycosides, this is not a valid practice for azithromycin or the macrolides. MICs of azithromycin and the macrolides must be compared with the infection site and phagocytic cell concentrations to determine the utility, or lack thereof, of one of these agents. Whereas azithromycin cellular penetration allows maximal pharmacodynamics potentially even against moderately or highly resistant pneumococci, the macrolides do so less optimally. Although there are no reports of widespread clinical failures resulting from macrolide/azalide resistance in pneumococci, it is expected that such reports will appear once the isolates become consistently highly resistant. This is likely to affect the macrolides, erythromycin and clarithromycin, before the azalide, azithromycin owing to the differences in pharmacokinetics of these drugs. Until then, it will be important to determine the MICs of not just one macrolide, but of all macrolides and azalides for the isolates. This will allow the clinician to make a pharmacokinetically and pharmacodynamically sound choice. By choosing clinical MIC breakpoints of 4-8 mg/L for oral macrolides and < or = 32 mg/L for oral azithromycin, rather than the present standard breakpoints, the clinician can make a macrolide/azalide choice that will optimize the pharmacodynamics of the drug against the isolated pathogen and result in the best possible clinical outcome. Once data concerning the cellular penetration of intravenous formulations of these drugs becomes available, it will be possible to develop clinical breakpoints for these formulations as well. Only through utilizing good antibiotic prescribing practices and by using the drugs appropriately when they are used, can resistance trends be stemmed. In this way, not only does a clinician treat the patient more effectively, but they also extend the antibiotic's useful life. PMID- 10459804 TI - Chemotherapeutic control of influenza. PMID- 10459805 TI - Identification and characterization of class 1 integrons in bacteria from an aquatic environment. AB - In a survey of 3000 Gram-negative bacteria isolated from an estuarine environment over a 2 month period, the incidence of class 1 integrons was determined to be 3.6%. Of 85 integrons studied further, 11 lacked both the qacEdelta1 and sull genes usually present in the 3' conserved segment of the integron. The qacEdelta1 and sull genes were identified in the 3' conserved segment of 36 integrons. The remaining 38 integrons lacked a sull gene but contained a qacE gene. The variable region of 74 integrons was characterized by PCR and sequence analysis. Forty of the integrons were found to lack integrated gene cassettes, although 21 of these 'empty' integrons were shown to contain inserted DNA which has been tentatively identified as a novel insertion sequence (IS) element. Of the 34 integrons which contained inserted gene cassettes, the aadA1a gene was found to be the most prevalent (74%). Nineteen integrons contained additional or other gene cassettes in their variable region, including those encoding resistance to trimethoprim (dfr1a, dfrIIc, dfrV, dfrVII, dfrXII), chloramphenicol (catB3, catB5), aminoglycosides (aadA2, aacA4, aacC1), beta-lactamases (oxa2) and erythromycin (ereA). This study confirms the occurrence of integrons in bacteria from a natural habitat and suggests that in the absence of continued antibiotic selective pressures, integrons which persist appear to preferentially exist without integrated antibiotic resistance gene cassettes. PMID- 10459806 TI - A variety of gram-positive bacteria carry mobile mef genes. AB - The mefE gene codes for a membrane bound efflux protein, which confers resistance to macrolides, and has been identified in Streptococcus pneumoniae. A variety of gram-positive organisms were examined. Twenty-six isolates of S. pneumoniae carried mefE and were resistant to erythromycin (MIC of 2-16 mg/L). Two additional isolates of Emr S. pneumoniae carried both ermB and mefE(MIC of 16-128 mg/L). One Micrococcus luteus, one Corynebacterium jeikeium, three Corynebacterium spp., two viridans streptococci and seven Enterocccus spp. also carried mef genes. It was possible to move the mef gene from all 11 S. pneumoniae tested to susceptible S. pneumoniae and/or Enterococcus faecalis recipients. The addition of DNase (1 g/L) did not affect the gene transfer. It was also possible to move the mef gene from donor Enterococcus spp., viridans streptococci, M. luteus, C. jeikeium and Corynebacterium spp. to E. faecalis recipients. Transconjugant isolates were resistant to erythromycin (MIC = 16 mg/L). Hybridization with a labelled mef oligonucleotide probe against Southern blots and bacterial dot blots confirmed the presence of the mef genes. This is the first time that a mobile mef gene has been identified in four different genera, from three distinct geographical locations. PMID- 10459807 TI - Rhodamine 6G efflux for the detection of CDR1-overexpressing azole-resistant Candida albicans strains. AB - We investigated the drug efflux mechanism in azole-resistant strains of Candida albicans using rhodamine 6G (R6G). No significant differences in R6G uptake were observed between azole-sensitive B2630 (9.02 +/- 0.02 nmol/10(8) cells) and azole resistant B67081 (8.86 +/- 0.03 nmol/10(8) cells) strains incubated in glucose free phosphate buffered saline. A significantly higher R6G efflux (2.0 +/- 0.21 nmol/10(8) cells) was noted in the azole-resistant strain (B67081) when glucose was added, compared with that in the sensitive strain B2630 (0.23 < or = 0.14 nmol/10(8) cells). A fluconazole-resistant strain C40 that expressed the benomyl resistance gene (CaMDR) also showed a low R6G efflux (0.16 +/- 0.06 nmol/10(8) cells) as did the sensitive strains. Accumulation of R6G in growing C. albicans cells was inversely correlated with the level of CDR1 mRNA expression. Our data also suggest that measurement of intracellular accumulation of R6G is a useful method for identification of azole-resistant strains due to CDR1-expressed drug efflux pump. PMID- 10459808 TI - Lentivirus-derived antimicrobial peptides: increased potency by sequence engineering and dimerization. AB - We have previously described a family of cationic amphipathic peptides derived from lentivirus envelope proteins that have properties similar to those of naturally occurring antimicrobial peptides. Here, we explored the effects of amino acid truncations and substitutions on the antimicrobial potency and selectivity of the prototype peptide, LLP1. Removal of seven residues from the C terminus of LLP1 had little effect on potency, but abrogated haemolytic activity. Replacement of the two glutamic acid residues of LLP1 with arginine resulted in a peptide with greater bactericidal activity. We discovered that the cysteine containing peptides spontaneously formed disulphide-linked dimers, which were 16 fold more bactericidal to Staphylococcus aureus. Monomeric and dimeric LLP1 possessed similar alpha helical contents, indicating that disulphide formation did not alter the peptide's secondary structure. The dimerization strategy was applied to magainin 2, enhancing its bactericidal activity eight-fold. By optimizing all three properties of LLP1, a highly potent and selective peptide, named TL-1, was produced. This peptide is significantly more potent than LLP1 against gram-positive bacteria while maintaining high activity against gram negative organisms and low activity against eukaryotic cells. In addition to new antimicrobial peptides, these studies contribute useful information on which further peptide engineering efforts can be based. PMID- 10459809 TI - Antibiotic susceptibility assay for Staphylococcus aureus in biofilms developed in vitro. AB - Four slime-producing isolates of Staphylococcus aureus were used in an antibiotic susceptibility assay for biofilms developed on 96-well polystyrene tissue culture plates. The study involved 11 antibiotics, two biofilm ages (6 and 48 h), two biofilm growth media (tryptone soy broth (TSB) and delipidated milk) and three antibiotic concentrations (4 x MBC, 100 mg/L and 500 mg/L). ATP-bioluminescence was used for automated bacterial viability determination after a 24 h exposure to antibiotics, to avoid biofilm handling. Under the conditions applied, viability in untreated biofilms (controls) was lower when biofilm growth was attempted in milk rather than in TSB. Various antibiotics had a greater effect on viability when used at higher (> or =100 mg/L) antibiotic concentrations and on younger (6 h) biofilms. Increased antibiotic effect was observed in milk-grown rather than TSB-grown biofilms. Phosphomycin and cefuroxime, followed by rifampicin, cefazolin, novobiocin, vancomycin, penicillin, ciprofloxacin and tobramycin significantly affected biofilm cell viability at least under some of the conditions tested. Gentamicin and erythromycin had a non-significant effect on cell viability. Transmission electron microscopy revealed that cells at the inner biofilm layers tend to remain intact after antibiotic treatment and that TSB grown biofilms favoured a uniformity of cell distribution and increased cell density in comparison with milk-grown biofilms. A reduced matrix distribution and enhanced cell density were observed as the biofilm aged. The S. aureus biofilm test discriminated antibiotics requiring shorter (3 h or 6 h) from those requiring longer (24 h) exposure and yielded results which may be complementary to those obtained by conventional tests. PMID- 10459810 TI - Effects of salicylate and related compounds on fusidic acid MICs in Staphylococcus aureus. AB - Salicylate, acetyl-salicylate, benzoate and ibuprofen increased fusidic acid MICs for fusidic acid-resistant and -susceptible strains of Staphylococcus aureus representing six genetic lineages. The effects of these substances on fusidic acid resistance levels occurred in a strain-dependent manner. The weak acid acetate, and acetaminophen did not alter fusidic acid resistance levels, while the addition of saligenin, the alcohol of salicylate, reduced gradient plate MICs for all strains studied. These findings indicate that a benzoic acid structure is required for the induction of increased intrinsic fusidic acid resistance levels. When 2 mM salicylate was added to media used in population analyses, the number of cells able to survive on high concentrations of fusidic acid increased. This increase in cell survival was observed in two unrelated fusidic acid-resistant strains, with chromosomal (WBG8287) or plasmid (WBG1576) mediated resistance determinants and two unrelated susceptible strains. The salicylate-induced increase in fusidic acid resistance was phenotypic at low fusidic acid concentrations (relative to resistance phenotype) for WBG8287 and a fusidic acid susceptible strain. On media containing salicylate and high fusidic acid concentrations, the mutation frequency to higher fusidic acid resistance levels was greater for WBG8287, compared with unsupplemented fusidic acid-containing media. These experiments provide evidence for a novel salicylate inducible fusidic acid resistance mechanism in S. aureus. PMID- 10459811 TI - In-vitro interactions of itraconazole with flucytosine against clinical isolates of Cryptococcus neoformans. AB - Treatment failures can occur in AIDS patients infected with Cryptococcus neoformans, despite aggressive antifungal therapy. Combination regimens with additive or synergic drugs could provide additional options for treating cryptococcosis. We studied the effects of itraconazole combined with flucytosine against 16 strains of C. neoformans var. neoformans. Combination therapy revealed different results for the various strains, including synergy (fractional inhibitory concentration (FIC) index 0.5, 63% of the interactions), addition (FIC >0.5 to 1.0, 31% of the interactions) and indifference (FIC >1.0 to <2.0, 6% of the interactions). Antagonism (FIC >2.0) was not observed. The efficacy of combination therapy was confirmed by quantitative cfu and killing curve assays. In particular, killing curves conducted in replicating cells showed that the addition of itraconazole prevented the development of flucytosine-resistant mutants of C. neoformans. These data show that the combination of itraconazole and flucytosine is significantly more active than either drug alone against C. neoformans in vitro. PMID- 10459812 TI - Flow cytometric assessment of amphotericin B susceptibility in Leishmania infantum isolates from patients with visceral leishmaniasis. AB - Amphotericin B susceptibility was measured by a flow cytometric membrane potential assay in Leishmania infantum promastigotes isolated from 11 immunocompetent children treated with liposomal amphotericin B and 19 HIV infected young adults treated with intralipid amphotericin B. Susceptibility levels were measured by the 90% inhibitory concentrations (IC90) representing the concentrations of drug that induced a 90% decrease in membrane potential compared with the control culture. In immunocompetent children, treatment was fully effective whatever the susceptibility of isolates to amphotericin B. In immunocompromised adults, on the contrary, unresponsiveness and relapses could be observed in all cases and IC90 increased in the course of successive treatments: a decrease of amphotericin B susceptibility in both promastigote and amastigote forms could be observed in a patient who had six relapses. These results suggest that the success of amphotericin B treatment depends greatly on patient immunity status, and indicate that successive relapses could enhance emergence of amphotericin B resistant isolates. The results demonstrate that the flow cytometric membrane potential assay can be used as an easy and reliable tool for studying the evolution of interactions between amphotericin B and the parasite membrane during long-term treatments. PMID- 10459813 TI - Pharmacological parameters of intravenously administered amphotericin B in rats: comparison of the conventional formulation with amphotericin B associated with a triglyceride-rich emulsion. AB - The LD50 determined in rats for the potent antifungal amphotericin B (AB) increased from 4.2 to 12.0 when the conventional AB-deoxycholate (DOC) was compared with AB associated with a triglyceride-rich emulsion (AB-emulsion). The reduction in amphotericin B toxicity is not due to a modification in plasma clearance, as both formulations seem to be removed from plasma at the same rate. Major differences in amphotericin B tissue distribution were not seen for kidney and liver but were seen for the lung. After 24 h administration of a single amphotericin B dose (2.0 mg/kg body weight) 23.78 +/- 11.71 mg/kg tissue was recovered from the lung of animals treated with AB-DOC whereas for AB-emulsion only 5.19 +/- 2.50 mg/kg tissue was recovered. The higher lethality of AB-DOC may be related to the higher concentration of amphotericin B in the lung. The therapeutic efficacy of AB-emulsion was similar to that of AB-DOC as attested by survival curves obtained after treatment of mice infected by Candida albicans. This is highly relevant, as the same is not necessarily found for other less toxic proposed vehicles. The equivalent efficacy and the increment in the LD50 will result in an important improvement in the therapeutic activity of amphotericin B. Furthermore, some data related to storage and stability indicate the clinical utility of this type of drug delivery. PMID- 10459814 TI - Antimicrobial treatment of an experimental otitis media caused by a beta lactamase positive isolate of Haemophilus influenzae. AB - A gerbil model of otitis media induced by a beta-lactamase producing and non serotypeable isolate of Haemophilus influenzae was used to assess the in-vivo efficacy of co-amoxiclav and cefuroxime at low (5 mg/kg) and high (20 mg/kg) doses. The MIC of the antibiotics tested against the pathogen was 1 mg/L (1/0.5 mg/L for co-amoxiclav). The organism was inoculated (+/-10(6) cfu) by transbullar challenge directly in the middle ear and antibiotic treatment was commenced 2 h post-inoculation and continued at 8 h intervals for three doses. Only high dose co-amoxiclav significantly reduced the number of culture-positive specimens as compared with untreated animals or with other treatment groups (91.7% as compared with 36.7% for high dose cefuroxime). The results obtained in any treatment group were related to middle ear antibiotic level/MIC. Antibiotic concentrations in the middle ear 90 min after administration were about 10% of serum levels at 15 min, probably related to a slight inflammatory response. Only after high dose co amoxiclav did the concentration in the middle ear exceed the MIC by a factor of four. In otitis media with effusion, if indicated, antibiotics active in vitro should be administered in high doses and, to avoid side effects, probably in short courses. PMID- 10459816 TI - Isepamicin in intensive care unit patients with nosocomial pneumonia: population pharmacokinetic-pharmacodynamic study. AB - A population approach was used to determine isepamicin pharmacokinetics in 196 intensive care unit patients treated for nosocomial pneumonia with isepamicin and a broad-spectrum beta-lactam. Patients were randomized in four groups with respect to the following isepamicin dosing regimens: (i) 15 mg/kg od for 5 days or (ii) 10 days, (iii) 25 mg/kg on the first day followed by 15 mg/kg od for 4 days or (iv) 9 days. A total of 1489 serum isepamicin concentrations were measured (median, eight per patient; range, 1-18). Mean +/- S.D. 1 h-peak levels at day 1 were 76 +/- 32 mg/L after the 25 mg/kg dose (n = 85) and 43 +/- 15 mg/L after the 15 mg/kg dose (n = 99). A bicompartmental model was fitted to the data by a mixed-effect modelling approach. Isepamicin clearance was related to age, bodyweight and serum creatinine level. Central volume of distribution was related to bodyweight. Pharmacokinetic parameters were independent of the dosage in the range 15-25 mg/kg and were not different in the patients treated for 5 or 10 days. Bayesian estimates of individual pharmacokinetic parameters were used to calculate various surrogate markers of isepamicin exposure to be tentatively correlated with clinical outcome and nephrotoxicity. No correlation was found between peak, AUC or their ratio with MIC and clinical efficacy. A weak correlation was found between the increase of serum creatinine level (day 1 versus day 5) and isepamicin 24 h trough level at day 1 (R2 = 0.10). These data do not favour a systematic therapeutic monitoring of isepamicin in intensive care unit patients, at least with the doses and antibiotic combinations used in this study. PMID- 10459815 TI - Treatment of experimental pneumonia in rats caused by a PER-1 extended-spectrum beta-lactamase-producing strain of Pseudomonas aeruginosa. AB - The antibacterial activity of imipenem, cefepime and piperacillin-tazobactam alone or in combination with amikacin against a Pseudomonas aeruginosa strain producing an extended-spectrum beta-lactamase (PER-1) were compared using an experimental model of pneumonia in non-leucopenic rats. Animals were infected intratracheally with 8.0 +/- 0.4 log10 cfu of P. aeruginosa, and therapy was initiated 3 h later, by which time animal lungs showed bilateral pneumonia containing >7 log10 P. aeruginosa cfu/g of tissue. Since rats eliminate antibiotics much more rapidly than humans, renal impairment was induced in all animals to simulate the pharmacokinetic parameters of humans. MICs determined using an inoculum of 4 log10 cfu/mL were as follows: imipenem, 1 mg/L; cefepime, 8 mg/L; piperacillin-tazobactam, 32 mg/L; and amikacin, 16 mg/L. A noticeable inoculum effect was observed with the four antimicrobial agents tested, which was greatest for cefepime and piperacillin-tazobactam. In-vitro studies indicated that imipenem was the beta-lactam with the greatest bactericidal effect and that amikacin was synergic only in combination with cefepime and imipenem. Cefepime and piperacillin-tazobactam alone failed to decrease bacterial counts in the rats' lungs 60 h after therapy onset, whereas imipenem and, to a lesser extent, amikacin significantly reduced the number of viable microorganisms. Combination of amikacin with any of the three beta-lactams tested was synergic, despite a high amikacin MIC for the infecting strain. These results paralleled our in-vitro data showing a marked inoculum effect for cefepime and piperacillin-tazobactam. Based on the results of this study, the best treatment for infections caused by this type of extended-spectrum beta-lactamase-possessing strain would be imipenem plus amikacin. PMID- 10459817 TI - Effects of voriconazole on Candida glabrata in vitro. AB - The effects of voriconazole on the growth, morphology and lipids of Candida glabrata were studied. MIC data showed that voriconazole was up to 32- to 64-fold more active than fluconazole in its ability to inhibit various C. glabrata strains. Voriconazole inhibited the growth of C. glabrata in a dose-dependent fashion. Electron microscope examination showed that voriconazole treatment affected the external and internal morphology of C. glabrata. Treatment of C. glabrata with voriconazole inhibited ergosterol synthesis and led to accumulation of methylated sterols. In contrast, no significant difference in phospholipid composition was observed between treated and untreated cells. PMID- 10459818 TI - Antimicrobial resistance patterns in urinary isolates from nursing home residents. Fifteen years of data reviewed. AB - The antibiotic resistance patterns of gram-negative bacteria isolated from nursing home patients between 1983 and 1997 were analysed. Escherichia coli was the most prevalent isolate (48%) followed by Proteus spp. (26%) and other Enterobacteriaceae (20%). During the study period, the susceptibility of E. coli decreased for co-trimoxazole (79% to 62%), increased for nitrofurantoin (79% to 91%) and remained unchanged for amoxycillin (41%). Susceptibility to norfloxacin, available from 1990, decreased from 87% to 71%. Similar trends were observed when the susceptibilities of all gram-negative urinary pathogens were combined. The changes in susceptibility can probably be attributed to the empirical prescribing practices in the nursing homes studied. PMID- 10459819 TI - Incidence and clinical impact of fluoroquinolone-resistant Escherichia coli in the faecal flora of cancer patients treated with high dose chemotherapy and ciprofloxacin prophylaxis. AB - This study evaluated the susceptibility of Escherichia coli to quinolones in 72 stool samples collected from 31 patients with solid tumours who had undergone high dose chemotherapy (HDC) and peripheral blood stem-cell (PBSC) rescue with ciprofloxacin prophylaxis. Samples were obtained at admission, after completing prophylaxis and three months later. All E. coli strains isolated from baseline samples were susceptible to quinolones. Fluoroquinolone-resistant E. coli strains were isolated in 10 (32%) patients in the second sample. In eight of these patients isolates were susceptible 3 months later. No patient developed infection due to fluorquinolone-resistant E. coli. No differences were observed in outcome between patients with susceptible and resistant flora. PMID- 10459820 TI - Glycopeptide tolerance in bacteria causing endocarditis. AB - Glycopeptides have been recommended as therapy for endocarditis. MICs and MBCs of vancomycin and teicoplanin were compared for 100 isolates from patients with proven bacterial endocarditis. MICs were generally lower for teicoplanin and tolerance to both agents was common. Almost all isolates of enterococci were tolerant to both glycopeptides. Among the streptococci, 78% were tolerant to teicoplanin and 57% to vancomycin. Similar findings were demonstrated for staphylococci. Although isolates appear sensitive to glycopeptides, bactericidal activity cannot always be predicted. If a glycopeptide is indicated for treatment of endocarditis, combination therapy with a suitable aminoglycoside should be considered unless MBC testing can be performed. PMID- 10459821 TI - Influence of Sanguisorba officinalis, a mineral-rich plant drug, on the pharmacokinetics of ciprofloxacin in the rat. AB - The significance of an interaction between ciprofloxacin and Sanguisorba officinalis L. (SO), a mineral-rich herbal medicine, was evaluated in this study. Male Sprague-Dawley rats (220-250 g) receiving ciprofloxacin dosages of 20 mg/kg po were concomitantly dosed with an aqueous extract of SO (equivalent to 2 g/kg crude drug). Blood and urine samples were collected over 6 and 24 h, respectively, for the quantitation of ciprofloxacin by HPLC. The presence of SO reduced significantly (P < 0.05) the maximum plasma concentration, the area under the concentration-time curve and the urinary recovery of ciprofloxacin, by 94%, 78% and 79%, respectively, compared with rats receiving only ciprofloxacin. The presence of SO also caused an eight-fold and two-fold increase in drug distribution (Vd, lambda(z)/F) and terminal elimination half-life (t1/2, lambda(z)) from 30.8 L/kg and 1.96 h, respectively. Therefore, should the use of both agents be required, sufficient time should be allowed to ensure the efficacy of ciprofloxacin. PMID- 10459822 TI - Efficacy and safety of an intravenous induction therapy for treatment of disseminated Mycobacterium avium complex infection in AIDS patients: a pilot study. AB - Monotherapy with macrolides for the treatment of disseminated Mycobacterium avium complex (MAC) bacteraemia leads to drug resistance and relapse of bacteraemia. Gastrointestinal intolerance is a common reason for treatment withdrawal of multidrug regimens. We have assessed the efficacy and safety of initial parenteral therapy together with a macrolide, for disseminated MAC infection, defined as two positive blood cultures, in AIDS patients. Patients received a daily infusion of amikacin 15 mg/kg + ethambutol 20 mg/kg + ciprofloxacin 400 mg/day, for 1 month, together with a macrolide by oral route. Fifteen patients were included and 13 (86%) achieved negative culture before the end of parenteral therapy. PMID- 10459823 TI - Emergence of mefA and mefE genes in beta-haemolytic streptococci and pneumococci in France. PMID- 10459824 TI - Carbapenem-hydrolysing beta-lactamase from clinical isolates of Pseudomonas aeruginosa in Portugal. PMID- 10459825 TI - Spectrophotometric determination of sparfloxacin in tablets. PMID- 10459826 TI - Acute pancreatitis with severe lactic acidosis in an HIV-infected patient on didanosine therapy. PMID- 10459827 TI - Do insulin-like growth factors mediate the effect of alcohol on breast cancer risk? AB - Despite a large number of epidemiologic studies demonstrating an increased risk of breast cancer in association with alcohol consumption, a causal relationship between alcohol intake and breast cancer risk remains to be determined. Several biological mechanisms have been proposed, but none of them explains well the features of the association, i.e. a modest increase in risk, a limited range of dose-response relationship and no further increase in risk among heavy drinkers. A new mechanism underlying a possible biological role of alcohol in breast cancer is proposed in this paper. Moderate consumption of alcohol increases the production of insulin-like growth factors (IGFs) by the liver and elevated IGFs via circulation stimulate or promote the development and/or growth of breast cancer. The effect of alcohol on IGF production declines among heavy drinkers as alcohol-caused liver-function damage results in no further increase in IGF production. Therefore, compared to moderate drinkers, heavy alcohol users do not have a higher risk of breast cancer. PMID- 10459828 TI - Why do we speak with the left hemisphere? AB - Human language tends to be associated with circuitry in the left cerebral hemisphere, regardless of individual hand dominance. This may have resulted from the coevolution of language and dexterous manipulation, specifically the use of the dominant hand to direct forces and point to objects in the environment. Asymmetric manipulation of physical objects reflects a fundamental asymmetry of perceptual-motor brain circuitry, which in turn results from the nature of the mechanical interaction between the organism and its environment. The natural selection of more effective manipulative ability, especially in the form of tool use, strengthened the fundamental organismal asymmetry, leading to distinct handedness and hemispheric dominance for manipulation and language in humans. The related subject of ocular dominance is also discussed. PMID- 10459829 TI - Induction of tolerance to hepatitis B virus: can we 'eat the disease' and live with the virus? AB - Hepatitis B virus (HBV) is a major health problem, with over 300 million HBsAg carriers worldwide. The HBV itself is non-cytopathic, and it is widely accepted that the mechanism of hepatocellular injury is the host anti-viral immune response. Current treatments, including the newly developed therapeutic modalities, are either based on antiviral drugs or focus on attempts to augment the anti-viral immune response. The results of these approaches have been largely disappointing. There is evidence, however, that subjects with a natural tolerance to HBV or a down-regulated immune response develop little or no liver injury, despite chronic viremia. Lately, it has been shown that it is possible to induce tolerance toward viruses by oral administration of major viral structural proteins. Here, we discuss the pathogenesis of HBV-mediated liver disease and approaches to down-regulate the immune response directed against liver cells by orally inducing tolerance toward the virus. We hypothesize that this acquired tolerance should turn chronic active hepatitis patients with deteriorating disease into 'healthy' virus carriers. The proposed new treatment strategy would redirect the focus from augmenting anti-viral immune response to inducing host tolerance towards the virus. PMID- 10459830 TI - Progression of cancer. AB - Carcinoma constitution originates in the hetero-duplication mitoses (4) which divide non-maturable stem cells into two different types of daughter cells: maturable and non-maturable cancerous stem cells. Throughout the progression of such mitoses, the organoid continuity of the cancer depends only on the non maturable daughter stem cells, not on the maturable ones. This type of cancer can be considered non-progressive or to be in the incubation stage, as it only enables cancer tissue to preserve its organoid continuity without allowing progressive growth. However, when an occasional mitotic phase of the hetero duplication mitotic progression undergoes genuine cell-phase duplication mitosis, this non-progressive or incubation stage of cancer is converted to a progressive type of cancer. This conversion is dependent on the reappearance of the complete mitotic condition of the mitotic maturation promoting system (MMPS) containing an abnormal supplement. Thus, a thorough investigation of the abnormal supplement and the reappearance of the complete mitotic condition is an essential part of research efforts to prevent and eradicate cancer. PMID- 10459831 TI - Upper gastrointestinal bleeding: an ammoniagenic and catabolic event due to the total absence of isoleucine in the haemoglobin molecule. AB - Upper gastrointestinal bleeding causes increased urea concentrations in patients with normal liver function and high ammonia concentrations in patients with impaired liver function. This ammoniagenesis may precipitate encephalopathy. The haemoglobin molecule is unique because it lacks the essential amino acid isoleucine and has high amounts of leucine and valine. Upper gastrointestinal bleeding therefore presents the gut with protein of very low biologic value, which may be the stimulus to induce a cascade of events culminating in net catabolism. This may influence the function of rapidly dividing cells and short half-life proteins. We hypothesize that, following a variceal bleed in a cirrhotic patient, the lack of isoleucine in blood protein is the cause of the exaggerated ammoniagenesis and catabolism. We propose that intravenous administration of isoleucine may serve as a simple therapeutic that transforms blood protein in a balanced protein, resulting in only a short-lived rise in ammonia and urea production, and preventing interference with protein synthesis. PMID- 10459832 TI - On ring chromatids. AB - The paper studies mammalian chromatids conformed as donut-like rings. Albeit enacting many topologic and dynamic aspects of straight chromatids (e.g. replication, eversion, antipodal segregation at anaphase, etc.), rings differ in certain properties regarding in particular disposal of soliton-like, kinked frustrations of their chirality during recurrent sister chromatid exchanges (SCEs). PMID- 10459833 TI - Re-entry into the cell cycle: a mechanism for neurodegeneration in Alzheimer disease. AB - Several recent findings demonstrated increased expression of cell cycle-related proteins in the degenerating neurons found in Alzheimer disease. We hypothesize that this apparent attempt to re-enter the cell cycle is a neuronal response to external growth stimuli that leads to an abortive re-entry into the cell cycle. However, since neurons of adults apparently lack the capacity both to divide in vivo and in vitro, it is possible that they lack the components necessary to complete the cell division process. Nonetheless, the importance of these findings is that they provide an explanation for the increased phosphorylation of cytoskeletal proteins such as tau and neurofilaments that represent the most striking intracellular changes in the disease. Further, it is our contention that inappropriate reentry into the cell cycle and interrupted mitotic processes are significant factors not only in the cytoskeletal pathology but also in the neuronal degeneration that characterizes the pathology of Alzheimer disease. PMID- 10459834 TI - Cysteine, glutathione (GSH) and zinc and copper ions together are effective, natural, intracellular inhibitors of (AIDS) viruses. AB - Sufficient essential nutrients such as methionine, cysteine, copper, selenium, zinc and vitamins C and E are indispensable for the maintenance of optimal (immune) cell functions. Parasitic organisms such as protozoa, fungi, bacteria and viruses also depend on these essential nutrients for their multiplication and functioning. An evolutionarily developed optimal distribution of available nutrients between host (cells) and parasitic organisms normally prevents diseases, the nature of which will depend on genetic and environmental factors. The way in which the right amount of cysteine, glutathione (GSH), and copper and zinc ions made available in the right place at the right time and in the right form can prevent an unchecked multiplication of (AIDS) viruses in a more passive or active way forms the basis for the AIDS zinc-deficiency hypothesis (A-Z hypothesis) presented in this article. Zinc and copper ions stimulate/inhibit/block in a concentration-dependent way the (intracellular) activation of essential protein-splitting enzymes such as HIV proteases. Zinc and copper ions as 'passive' virus inhibitors. Apart from this, zinc ions directly or indirectly regulate, via zinc finger protein molecular structures, the activities of virus-combating Th-1 cells such as cytotoxic T-cells (CTLs). Zinc ions as regulators of the active, virus-combating Th-1 cells. Zinc and copper ions that remain available in sufficient amounts via cysteine/GSH are effective natural inhibitors/combaters of (AIDS) viruses and thereby prevent the development of chronic virus diseases that can lead to AIDS, autoimmune diseases, (food) allergies and/or cancer. A safe, relatively inexpensive and extensively tested medicine such as N-acetylcysteine (NAC) can help in supplying extra cysteine. The anti-HIV peptide T22, synthesized on the basis of two natural peptides from the Tachypleus tridentatus and Limnus polyphemus crabs, appears to be able to serve as supplier/carrier molecule of cysteine and zinc and/or to hinder the entry of HIVs into cells by way of the CD4 receptor. PMID- 10459835 TI - The biographical component of schizophrenia: a two-faced definition of relationship? AB - The schizophrenic's disturbed relationship with reality is generally ascribed to a mental illness of endogenous origin. Following the author's paper on a fixation disorder (1) provable in these patients, she expounds that their disturbed relationship with reality does not primarily concern their consciousness of objective reality but of their own self. As a result of contradictory definitions of relationship within his group (2,3) the schizophrenic is unable to integrate the awareness of his potentialities with the awareness of his social role. He cannot protect himself against a joker role imposed on him since his fixation disorder acts like a one-way valve impeding him from taking an active part in the definition of relationship. A hypothesis is presented as to how the schizophrenic's described fixation disorder may cause this one-way functioning of the definition of relationship. PMID- 10459836 TI - Asymmetric DDE (D35E)-like sequences in the RAG proteins: implications for V(D)J recombination and retroviral pathogenesis. AB - Experimental evidence suggests that the mechanism of vertebrate V(D)J recombination catalyzed by the vertebrate RAG proteins is similar to both retroviral integration and the transposition of IS630/Tc1-family transposons. The mechanism of both retroviral integration and IS630/Tc1 element transposition is well characterized and utilizes a functional metal ion binding site termed the DDE (or D35E) motif. We have previously identified a DDE-like region in the RAG-2 protein and a similar region within the RAG-1 protein. In this work, we propose that interference between DDE-like regions in the RAG proteins and the DDE-site of the HIV integrase may be a mechanism of retroviral pathogenesis in cells in which both the RAG proteins and retroviral integrase are co-expressed. PMID- 10459837 TI - Differences in radiosensitivity between brain, small and large arteriovenous malformations. A tentative explanation of the incongruent results of radiotherapy. AB - Intriguing differences in the results of brain arteriovenous malformation (AVM) therapy with ionizing radiation are reported in the literature, i.e. AVMs obliterate at different time intervals from the initiation of radiotherapy although presenting with similar nidus volumes that are treated with the same radiation dose. The purpose of this paper is to present a new concept that explains the variety of results after radiotherapy. To account for an individual AVM's responsiveness to radiotherapy, four variables have been identified from the literature. The variables are factors that are generally accepted to influence the biological effect of radiation. Combined, they can visualize a variety of AVM tissues as well as surrounding normal brain. These hypothesized variations in the type of tissues are then matched with the incongruent results of radiotherapy in order to clarify the issue. Future clinical research programs and possible therapeutical implications of this concept are proposed and discussed. PMID- 10459838 TI - Aluminium-induced biphasic effect. AB - Increased bioavailability of aluminium has raised concerns about the toxic effect of aluminium. The cholinotoxic effect of aluminium is already well established. The biological response of an organism following exposure to a chemical may be biphasic. Although aluminium-induced biphasic change has been reported in diverse organ systems, the biphasic effect on cholinergic system has received less attention. In vitro and in vivo studies have demonstrated an aluminium-induced biphasic effect on the marker enzyme of cholinergic system, acetylcholinesterase. The biphasic effect of aluminium on the acetylcholinesterase enzyme activity may be due to the direct neurotoxic effect of the metal and the level of aluminium accumulated. Among various hypotheses, peroxidation-induced changes in the structure of membrane following aluminium accumulation seems to explain the biphasic effect of aluminium on acetylcholinesterase activity. PMID- 10459839 TI - Covariance of breast cancer incidence with smoking-, oestrogen- and diet-related cancers in pre- and postmenopausal women in Sweden. AB - Effects of smoking on breast cancer risk remains controversial. Tar products have been claimed to increase risk, antioestrogenic effects to reduce risk. Another possibility is that associations to smoking have been confounded by diet. The increasing incidence of breast cancer from 1960 to 1994 in Sweden is parallel to that of lung cancer and the increasing proportion of female smokers. The incidence of endometrial and colon cancer was in premenopausal women negatively and in postmenopausal women positively related to the incidence of breast cancer. Possible explanations and hypotheses to the different co-variance between breast cancer and lung, endometrial and colon cancer in pre- and postmenopausal women are discussed from the perspectives of smoking, sex hormones and diet. It is concluded that the strong and specific positive relationship between breast and lung cancer in premenopausal women is compatible with the hypothesis that aromatic hydrocarbons may be involved in the causation of disease. PMID- 10459840 TI - Sudden infant death syndrome: hypothalamic failure to sense elevated blood pyrogens. AB - Sudden infant death syndrome (SIDS) is frequently associated with a mild infection, the incidence peaking during the third month of life. We hypothesize that the neonatal immaturity of both the acute febrile response and hypothalamus promote neonatal protection from SIDS. Vagal afferents modify the febrile response. Vagotomized rodents displayed a loss of febrile responsiveness in a 'non-sensing' brain. The failure of a 'non-sensing' brain to react to elevated blood pyrogens leads to failure of the febrile response and to a shock-like state. SIDS infants may appear well yet, within hours of this observation, may be found dead. There is a mismatch between the acute febrile response and hypothalamic hypoactivation. The discrepancy increases with development. There is an elevated cytokine response in endothelial cells which induces nitric oxide (NO) production and retarded development of the hypothalamus. Cigarette smoke also induces NO production and retards hypothalamic development by augmented apoptosis. Zinc inhibits this effect in mouse thymocytes. Fetal haemoglobin (HbF) induces hypoxia, which is a stimulator of the immune response while vasodilator gases (carbon monoxide (CO), NO) reduce hypothalamic function. The hypothalamic failure to sense elevated blood pyrogens induces toxic shock - a feature of SIDS. PMID- 10459841 TI - Sudden infant death syndrome: oxidative stress. AB - In studies of oxidative stress in sudden infant death syndrome (SIDS) there were two major findings: (1) During normal post-natal development, there was a gradual decline in the number of Cu/Zn superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) immunoreactive neurons in the hippocampus and parahippocampus gyrus in the brain; (2) The total number of immunoreactive neurons was elevated in SIDS victims compared to age-matched controls in infants 6 months of age and under (1). SOD and neuronal aging and degeneration in the hippocampus and neocortex were features of SIDS, Alzheimer's disease and Down's syndrome. In the SIDS study of infants from 3-6 months of age, the elevation of SOD in SIDS victims was significant, whereas no significant elevation of GSHPx was detected. An imbalance between SOD and GSHPx was said to be crucial in the prevention of toxicity of free radicals (1). Zinc-deficient cells cannot up-regulate gene expression of the scavenger enzymes SOD and GSHPx in cells exposed to high levels of superoxide and hydrogen peroxide (2). GSHPx coupled to reduced nicotine adenine diphosphate (NADPH) regenerating systems via glutathione reductase is virtually able to guarantee an effective protection of biological structures against oxidative attack (22). When the capacity of the cell to regenerate GSH is exceeded - primarily due to an insufficient supply of NADPH-oxidised glutathione (GSSG) is released from the cell and protein synthesis turns off (20). We hypothesize that the increased incidence of aging and neuronal death and increased incidence of SOD and GSHPx reactive neurons in early post-natal development indicates an increased up-regulation of gene expression of scavenger enzymes during high exposure to oxidative stress after birth. GSH-dependent peroxide metabolism is linked to the pentose phosphate shunt via NADPH-dependent glutathione reductase (GR). GSHPx is a selenium containing enzyme which together with catalase (CAT) SOD and vitamin E protects cells in the free radical chain. Zinc upregulates gene expression of these antioxidants. PMID- 10459842 TI - A possible link between neural tube defects and ultraviolet light exposure. AB - The protective role of folate in preventing neural tube defects is now well established. The hypothesis is advanced here that photolysis of folate by ultraviolet (UV) light may, in some women, precipitate a folate deficiency sufficient to cause a neural tube defect (NTD) during the first few weeks of pregnancy. This hypothesis is supported by the demonstration of in vitro photolysis of folate by simulated strong sunlight (1), a decline in folate levels in light-skinned subjects exposed to UV light for dermatological conditions (1), and the occurrence of NTDs in the offspring of women who exposed themselves to high levels of UV light on the sunbeds of tanning salons (2). If established, a connection between in vivo folate photolysis by UV light, clinical folate deficiency and NTDs would suggest that intense or prolonged periconceptual exposure of women to UV light for recreational or therapeutic reasons should be avoided. PMID- 10459843 TI - Tuberculosis I: a conceptual frame for the immunopathology of the disease. AB - An analysis of the cellular and humoral immune responses after bacille Calmette Guerin (BCG) vaccination and during tuberculosis treatment favors the hypothesis of an immune defence developed in four overlapping successive stages. The initial immune response is innate. The following two intermingle innate and specific responses against low molecular weight oligopeptidic and nonpeptidic antigens, as muramyldipeptide and trehalose dimycolate, and large molecular weight nonpeptidic antigens such as lipoarabinomannan. The ultimate specific response is directed against protein antigens as Antigen 60. BCG and primary tuberculosis (TB) infections induce cellular and humoral immune responses essentially against oligopeptidic and small and large molecular weight nonpeptidic antigens. Immune responses against non-peptidic substances contribute to the immunoprotection of the infected person who develops a primary infection. Some infected people allow the expression of the immunosuppressive activity of the pathogen. This results in the synthesis of interleukin-10 (IL-10), which suppresses the formation of interferon-gamma (INF-gamma) and IL-2, and of IL-6, which suppresses T-cell responses. These patients have a skewed immune response against non-peptidic antigens and present with symptoms. They will not recover unless responses directed against proteinic antigens occur, which restore INF-gamma and IL-2 production. The formation of immumoglobulin-G (IgG)-type antibodies and of a cellular immunity against mycobacterial peptidic antigens is essential for a good protection against a post-primary infection. PMID- 10459844 TI - HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) pathogenesis hypothesis. A shift of homologous peptides pairs, central nervous system (CNS)/HTLF-1, HTLV-1/thymus, thymus/CNS, in a thymus-like CNS environment, underlies the pathogenesis of HAM/TSP. AB - Determinants shared by thymus, brain and HTLV-1 induce lymphocytic neurotropism and demyelinization in HAM/TSP, within the framework thymus-like brain environment. The disease evolves in two phases. The first phase of the disease would be dependent on CD4 T-lymphocytes specific for thymic autoantigens, reactivated by viral antigens homologous to thymus and CNS autoantigens. During this phase, demyelinization could be due initially to a stop in the synthesis of myelin following an altered expression of adhesion proteins at the surface of oligodendrocytes and neurons. The second phase, which covers the inflammatory and chronic character of the disease, would be dependent, on the one hand, on CD8 T lymphocytes specific for viral peptides, and on the other hand, on CD8 T lymphocytes specific for peptides arising from the cell-proteases induced progressive proteolysis of protein components from the myelin layers and other protein components of the CNS. Non-specific inflammatory and non-inflammatory cytokines keep the activation going of the different cellular types. The thoracic spinal cord cell-location specificity would be linked to a peptidic coherence between HTLV-1 (significant agent), thymus and thoracic spinal cord antigens, genetically peculiar to HAM/TSP patients. PMID- 10459845 TI - Mycoplasma: a new potential vector for gene therapy? AB - The introduction of foreign genetic material in living cells is the basis of the current protocols of gene therapy. The concept that the de novo synthesis of a foreign therapeutic protein requires the entrance of the corresponding gene into target cells via virus or synthetic vectors is directly inherited from experiments on bacterial transduction or transformation. However, the difficulties inherent in the penetration and the expression of foreign DNA into eucaryotic cells are probably responsible for the low efficiency of this therapeutic approach. In this paper, we explore the possibility of avoiding these limiting critical steps by expressing the foreign gene on the surface rather than inside the target cells by the use of mycoplasma, the smallest reported living cell. The absence of transfer of genetic information between this vector and eucaryotic target cells, the sensibility of mycoplasmas to antibiotics and their cytadherance are among the interesting features of this potential vector. The interest of this new vector in the case, e.g. of the gene-directed enzyme prodrug therapy of solid tumors, is discussed. PMID- 10459846 TI - The 'anti-delirium' theory of electroconvulsive therapy action. AB - It is proposed that electroconvulsive therapy (ECT) is not specifically mood elevating or anti-depressant but that its effect is as an anti-delirium intervention. I suggest that ECT exerts its primary therapeutic effects by inducing a generalized epileptic seizure which operates on the brain like a deep and restorative sleep that acts rapidly to resolve delirium. Provided that the diagnosis is made using sufficiently sensitive criteria, delirium is here assumed to be a common feature of many so-called 'functional' psychoses - frequently occurring as a consequence of sleep deprivation, and leading to symptoms such as hallucinations, bizarre delusions and psychomotor retardation. Testable predictions of this 'anti-delirium' theory of ECT action are described. PMID- 10459847 TI - A new hypothesis of neurodegenerative diseases: the deleterious network hypothesis. PMID- 10459848 TI - Growth response of breast epithelial cells to estrogen is influenced by EGF. AB - Estrogen-induced growth stimulation has not previously been demonstrated in estrogen receptor (ER) cDNA transfected human cell lines in contrast to breast cancer cell lines expressing endogenous ER. On the contrary, estrogen usually inhibits cell growth of ER transfected cell lines. Growth inhibition by estrogen has also been demonstrated in our cell line, F9, which is an ER transfected subline of HMT-3522 breast epithelial cells derived from fibrocystic disease and propagated in chemically defined medium. By omitting EGF in the medium, we have demonstrated not only an increased transcriptional activity of the ER but also- after an adaptation period--estrogen-dependent growth of the cells, and we have succeeded in establishing a new subline, S3B, that requires 17beta-estradiol (E2) for growth. This is the first example of a nonmalignant, human breast epithelial cell line which is dependent on estrogen for continued growth. The S3B cells express functional ER as measured by transcriptional activity. ER-E2 induced transcription was not inhibited by EGF as in F9 cells. We propose that a growth stimulatory response of breast epithelial cells in vitro to E2 is dependent on an inactive or down-regulated EGF receptor signaling pathway and it is possible that the effect of estrogen on normal breast epithelium in vivo also is modulated by the EGFR. PMID- 10459849 TI - Activation of the insulin-like growth factor binding protein-1 promoter by progesterone receptor in decidualized human endometrial stromal cells. AB - Insulin-like growth factor binding protein-1 (IGFBP-1) is induced extensively when human endometrial stromal cells are decidualized by progestin and relaxin in a long-term primary culture system. The purpose of this study is to investigate whether progesterone receptor (PR) directly activates the IGFBP-1 gene promoter. In decidualized stromal cells, activity of the IGFBP-1 promoter (from -1.2 kb to +68 bp) containing putative progesterone-response elements (PREs) was increased 80-fold. Mutation of either 5' or 3' half-site of the putative PRE1 site (from 193 to -179 bp) reduced the promoter activity. Mutations that converted PRE1 closer to consensus PRE increased the promoter activity. In undifferentiated stromal cells, mutations of PRE sites had no effect on the promoter activity. When a PR expression vector (hPR1) was cotransfected, progestin increased promoter activity derived from p275CAT but not from p1.2CAT, suggesting that the function of PRE1 was repressed by the region from -1.2 kb to -275 bp in the promoter. Progestin did not increase promoter activity derived from p275CAT without cotransfection of hPR1, suggesting that endogenous PR alone is insufficient to activate PRE1. In summary, results indicate that the PRE1 site of the IGFBP-1 promoter mediates a direct activation of PR on transcription specifically in decidualized stromal cells. PMID- 10459850 TI - Discordant expression of insulin-like growth factors and their receptor messenger ribonucleic acids in endometrial carcinomas relative to normal endometrium. AB - The inappropriate expressions of insulin-like growth factors (IGF-I and II) and IGF-I receptor (IGF-IR) are implicated in the malignant growth of many cancers. To determine changes, if any, in the levels of expression of IGFs and IGF receptor genes in neoplastic endometrium, relative to normal endometrium, the mRNA levels of IGF-I and II and of IGF-IR and IIR were measured in samples of endometrial carcinomas (EC) and normal endometrium, through all phases of the menstrual cycle, by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) assays. In normal endometrium, the mRNA levels of IGF-I were elevated in the proliferative and early secretory phases. The IGF-II mRNAs were relatively high in the proliferative phase, but unaltered through early and late secretory phases. Significantly elevated levels of IGF-II transcripts were observed during the menstrual phase, suggesting a possible role of IGF-II in endometrial regeneration. A positive correlation between the levels of IGF-I and IGF-IR mRNAs, apparent in the samples of normal endometrium, was not observed in endometrial carcinomas. The IGF-IR and IIR mRNA levels were elevated in endometrial carcinoma samples. On the other hand, the IGF-I and II mRNA levels were conspicuously low in many carcinoma samples, which were not associated with hyperplasia (type II EC), but relatively elevated in two other carcinoma samples, associated with adenomatous hyperplasia (type I EC). These results albeit with few samples suggest the possibility that the overexpressed receptor, IGF-IR, could be activated differently in two types of endometrial carcinomas, namely ligand-dependently in type I ECs and ligand-independently in type II ECs. PMID- 10459851 TI - Retinoid and androgen regulation of cell growth, epidermal growth factor and retinoic acid receptors in normal and carcinoma rat prostate cells. AB - Recent in vivo and in vitro studies suggest that retinoic acid receptor (RAR) mediated processes may be involved in androgen regulation of prostate cells in a manner that may be altered during prostatic carcinogenesis. We tested this hypothesis in the newly established carcinoma and non-carcinoma rat prostate epithelial cell lines, NRP-154 and NRP-152, respectively. In DMEM/F-12 medium supplemented with 10% charcoal stripped fetal calf serum (cFCS), the number of both NRP-152 and NRP-154 cells were stimulated by testosterone (T), with a 4-fold greater effect in NRP-152 than in NRP-154 cells. Retinoic acid (RA) alone also stimulated the growth of NRP-152 cells, but failed to induce cell growth of NRP 154 cells. Importantly, the level of RAR alpha mRNA was elevated whereas the levels of RAR gamma and androgen receptor (AR) mRNA were lower in NRP-154 cells compared to those in NRP-152 cells. Treatment of NRP-152 cells with increasing doses of T resulted in a dose-dependent decrease and rebound of the level of RAR alpha and gamma mRNA in NRP-152 cells; these effects were not apparent, if not reversed, in NRP-154 cells. Both ligand binding and Western blot analyses revealed that epidermal growth factor receptor (EGF-R) was stimulated by 20 nM T but was suppressed by 0.1 microM RA, which also attenuated the stimulating effects of T on EGF-R in NRP-152 and to a lesser extent in NRP-154 cells. The differences in the level and androgen regulation of RAR mRNAs and reciprocal regulation of EGF-R expression by T and RA between NRP-154 and NRP-152 cells suggest that variations in the EGF-R and RAR signal events may contribute to differences in growth rate between these two cell lines. PMID- 10459852 TI - Participation of protein kinase C alpha in 1,25-dihydroxy-vitamin D3 regulation of chick myoblast proliferation and differentiation. AB - Changes in morphology and DNA synthesis in cultured myoblasts in response to 1,25 dihydroxy-vitamin D3 [1,25(OH)2D3] have previously suggested that the vitamin D hormone may affect muscle cell proliferation and differentiation. However, this interpretation was not substantiated by measurement of specific biochemical markers of myogenesis. To study the effect of 1,25(OH)2D3 on muscle development, chicken embryo myoblasts were cultured for 1-6 days in the presence or absence of 1,25(OH)2D3 (10(-9) M). The hormone increased DNA synthesis and decreased creatine kinase activity, indicating stimulation of cell proliferation and inhibition of myogenesis, in undifferentiated myoblasts (1 day of culture). At longer culture intervals, when myoblasts elongate and fuse to form differentiated myotubes, 1,25(OH)2D3 promoted myogenesis, as indicated by an inhibition of DNA synthesis and an increase in specific muscle differentiation markers as creatine kinase activity and myosin expression. The role of protein kinase C (PKC) in mediating the effects of hormone and the likely PKC isoform involved were also investigated. Increased PKC activity was observed during 1,25(OH)2D3 stimulation of myoblast proliferation whereas inhibition of PKC activity accompanied the effects of the hormone on myoblast differentiation. The specific PKC inhibitor calphostin suppressed hormone potentiation of DNA synthesis in proliferating myoblasts. 1,25(OH)2D3-dependent changes in the expression of PKC isoforms alpha, beta, delta, epsilon and zeta during myogenesis were investigated by Western blot analysis. The early stimulation of myoblast proliferation by the hormone mainly correlated to increased PKC alpha expression whereas decreased PKC alpha levels were observed during the subsequent activation of myoblast differentiation. These results support that 1,25(OH)2D3 has a function in embryonic muscle growth and maturation, and PKC alpha may participate in the signal transduction pathway which mediates the response to the hormone. PMID- 10459853 TI - Growth hormone and insulin-like growth factor-I enhance beta-glucuronidase gene activation by androgen in mouse kidney. AB - Beta-glucuronidase (GUS) is a lysosomal enzyme that, in mouse kidney, is subject to control by multiple hormones: androgen, which increases GUS transcription; estrogen, which antagonizes androgen-mediated stimulation of GUS; and growth hormone (GH), which appears to be necessary for the full androgen effect. Neither estrogen nor GH affects GUS in the absence of androgen. In hypophysectomized or pituitary dwarf mice the reduced androgen stimulation of GUS can be partially restored with GH treatment. Androgen-induced GUS mRNA increased significantly with intermittent GH, compared to no GH or continuous GH. Intact mice subjected to continuous infusion of GH showed a depressed androgen effect on GUS similar to that seen in GH-deficient mice. Thus, pulsatile GH is required for the full androgen response. Insulin-like growth factor-I (IGF-I) also restored GUS induction by androgen in GH-deficient mice. We conclude that GH enhances the effect of androgen on the GUS gene via IGF-I. Using transgenic mice, we have also identified a genetic variant of the GUS gene that is insensitive to GH enhancement of the androgen effect. PMID- 10459854 TI - Stimulation of specific binding of [3H]-progesterone to bovine luteal cell surface membranes: specificity of digitonin. AB - Non-genomic actions of progesterone have been described in the ovary, and luteal membranes of several species have been shown to possess specific binding sites for [3H]-progesterone. However, binding of radiolabelled progesterone to luteal membranes was demonstrable only in the presence of digitonin. Digitonin is a non ionic detergent which is thought to act by forming one-to-one complexes with certain sterols. It is also a cardiotonic agent, inhibiting (Na+-K+) ATPase activity by interaction with the extracellular (ouabain/K+) binding site. We therefore investigated which properties of digitonin were responsible for its stimulatory actions on progesterone binding to bovine luteal membranes. A range of compounds with detergent, cardiotonic and or cholesterol-complexing activities were tested for their effects on [3H]-progesterone binding to bovine luteal membrane fractions, and on haemolysis of rat erythrocytes. Stimulation of progesterone binding to luteal membranes was highly specific for digitonin, and a number of ionic and non-ionic detergents, cardenolides, saponins and cholesterol complexing reagents tested failed either to stimulate [3H]-progesterone binding to bovine luteal membranes in the absence of digitonin, or to inhibit binding specifically in the presence of digitonin. When digitonin was first reacted with excess cholesterol or pregnenolone to form the respective digitonides, stimulatory activity was greatly reduced, suggesting that the ability of digitonin to interact with (an) endogenous steroid(s) may be important in its action. High performance liquid chromatography (HPLC)-mass spectrometry of commercially available digitonin preparations indicated the presence of numerous minor impurities in most commercial digitonin preparations. Three major UV absorbing peaks were isolated and characterised by mass spectrometry: all stimulated progesterone binding to bovine luteal membrane receptors in a dose dependent manner, though to differing extents. Our data suggest that the unique action of digitonin on luteal membrane progesterone receptors is not related to its detergent or cardiotonic properties, but appears to be related to its ability to complex with membrane sterols. PMID- 10459855 TI - The HDQVH-motif in domain E of the estradiol receptor alpha is responsible for zinc-binding and zinc-induced hormone release. AB - The estradiol receptor alpha and proteolytic fragments thereof which contain the entire ligand-binding domain E, bind 65Zn with high affinity. Four putative double-histidine zinc-binding sequences can be identified within the hormone binding domain E: HDQVH [amino acid (aa) 373-377], HIH (aa 474-476), HFRH (aa 513 516) and HRLH (aa 547-550). Only the HDQVH-motif is responsible for the 1:1 zinc binding to domain E because the proteolytic (endo-Lys-C) 17 kDa fragment (aa 303 467) from porcine estradiol receptor alpha possesses the zinc-binding ability but none of the fragments containing the other motifs. In addition, H373A- and H377A mutants lack the metal-binding capacity. Moreover, divalent metal ions are able to release estradiol out of the binding-niche. The order for this feature parallels the competition pattern of 65Zn-binding: Mg2+ < Ni2+ << Zn2+ < or = Cu2+. Mutant estradiol receptor alpha fragments (H373A and H377A) lack the zinc induced hormone release. PMID- 10459856 TI - Lack of responsiveness to TGF-beta1 in a thyroid carcinoma cell line with functional type I and type II TGF-beta receptors and Smad proteins, suggests a novel mechanism for TGF-beta insensitivity in carcinoma cells. AB - Transforming growth factor-beta (TGF-beta) is a multifunctional cytokine. In the present study we have investigated the expression of TGF-beta receptors (TbetaR's) and SMAD proteins in non-neoplastic and neoplastic thyroid follicle cells. We found expression of all TbetaR's (type I, II and III) and SMAD proteins analysed (Smad2, Smad3, Smad4, Smad6 and Smad7). Five out of six human anaplastic thyroid carcinoma cell lines were growth inhibited by addition of TGF-beta1, and therefore considered to be TGF-responsive. One cell line however, HTh 7, did not respond to TGF-beta1 with growth inhibition, induction of the extracellular matrix protein fibronectin or immediate early genes junB, Smad6 and Smad7 mRNA. Analysis of the TGF-beta intracellular signalling pathway in HTh 7 cells showed that receptors were capable of signalling, e.g. Smad2 phosphorylation and SMAD nuclear translocation. In summary, our data shows abundant expression of TGF-beta signalling components in thyroid follicle cells, and the escape from TGF-beta sensitivity in one anaplastic thyroid carcinoma despite an apparently functional TGF-beta/SMAD-signalling pathway, indicating a novel mechanism for TGF-beta insensitivity. PMID- 10459857 TI - The first exon of the human sc gene contains an androgen responsive unit and an interferon regulatory factor element. AB - Secretory component (SC) plays a key role in the transport of IgA and IgM to the lumina of many glands. The gene is constitutively expressed, but can be modulated by hormonal and immunological stimuli. Recently, the promoter and the first exon of the human sc gene have been cloned. The first exon contains a putative androgen/glucocorticoid response element (ARE/GRE) and an Interferon Regulatory Factor Element (IRF-E). Here we show that the ARE/GRE can bind the DNA-binding domain (DBD) of both the androgen (AR) and glucocorticoid receptor (GR) with a preference for the AR-DBD. In transient transfection experiments, this element confers higher responsiveness to androgens than to glucocorticoids. The IRF-E can function as an IRF-2, but surprisingly not as an IRF-I responsive element. We postulate that these two regulatory elements play a key role in the complex regulation of the sc gene in vivo. PMID- 10459858 TI - Nitric oxide inhibits human aldosteronogenesis without guanylyl cyclase stimulation. AB - Deta nonoate (deta-NO), a zwitterion nitric oxide (NO) donor, potently inhibited forskolin- and angiotensin II-stimulated aldosterone production in human adrenocortical H295R cells in a concentration-dependent manner (0.1-1000 microM). The half-maximal and maximal inhibition of forskolin-evoked aldosteronogenesis occurred at 0.6 and 100 microM deta-NO, respectively. The respective half-maximal and maximal deta-NO-mediated inhibition of angiotensin II-stimulated aldosterone generation occurred at 150 microM and 1 mM. In H295R cells, deta-NO and sodium nitroprusside did not stimulate cGMP production, and the soluble guanylyl cyclase inhibitor oxadiazoloquinoxalinone (10 microM) did not block deta-NO-mediated attenuation of aldosteronogenesis. 25-Hydroxycholesterol (10 microM)-facilitated aldosterone synthesis was also diminished with half-maximal and maximal inhibition occurring at 120 microM and 1 mM deta-NO, respectively. Taken together, these results demonstrate that NO inhibits human aldosteronogenesis without stimulating guanylyl cyclase in H295R cells. PMID- 10459859 TI - Osteogenic protein-1 increases gene expression of vascular endothelial growth factor in primary cultures of fetal rat calvaria cells. AB - Osteogenic protein-1 (OP-1 or BMP-7) stimulates new bone formation in vivo and induces cell proliferation and differentiation of osteoblasts in vitro. In the present study, we examined effects of OP-1 on the expression of vascular endothelial growth factor (VEGF) in primary cultures of fetal rat calvaria (FRC) cells. OP-1 increased the steady-state level of VEGF mRNA by about 3-fold in an OP-1 concentration- and time-dependent manner. The increase in VEGF mRNA level depended on transcription and was sensitive to cell replication. The VEGF mRNA stability was unaffected. The mRNA levels for both types of VEGF receptors, Flk-1 and Flt-1 were low but detectable in FRC cells by RT-PCR and were not changed by OP-1. Inhibition of VEGF synthesis and function by antisense oligonucleotide and by suramin, respectively arrested the OP-1-induced alkaline phosphatase activity and mineralized bone nodule formation. Together with published studies of VEGF on vascular endothelial cells which are usually found in close proximity to osteoblastic cells in vivo, these results suggest that VEGF participates in the OP-1-induced osteogenesis by taking part in bone cell differentiation and by promoting angiogenesis at the site of bone formation. PMID- 10459860 TI - Transplantation of primary bovine adrenocortical cells into scid mice. AB - Bovine adrenocortical cells were transplanted into scid mice, using a small cylinder inserted beneath the kidney capsule. The tissue formed from primary bovine adrenocortical cells replaced the essential functions of the animals' own adrenal glands, which were removed during the cell transplantation procedure. Most adrenalectomized animals bearing transplanted cells survived indefinitely, whereas adrenalectomized control animals died following surgery. Formation of well-vascularized tissue at the site of transplantation was associated with stable levels of cortisol in the blood, replacing the mouse glucocorticoid (corticosterone). Ultrastructurally, the cultured cells before transplantation had characteristics of rapidly growing cells, but tissue formed in vivo showed features associated with active steroidogenesis. We investigated two potentially critical aspects of the procedure: the provision of support for angiogenesis in the transplant by the inclusion of FGF-secreting 3T3 cells with the adrenocortical cells; and the administration of synthetic steroids as a temporary replacement for steroids lost by adrenalectomy. We found that FGF was required for the rapid formation of well-vascularized tissue, whereas steroid administration avoided some early mortality but was not absolutely required. In contrast to transplants formed from clonal cells, which did not usually secrete aldosterone, transplants formed from primary bovine adrenocortical cells, even though derived from the zona fasciculata, secreted aldosterone as well as cortisol. PMID- 10459861 TI - Co-ordinate expression of activin A and its type I receptor mRNAs during phorbol ester-induced differentiation of human K562 erythroleukemia cells. AB - Activins were originally isolated based on their ability to stimulate follicle stimulating hormone secretion but later they have been shown to regulate a number of different cellular functions such as nerve cell survival, mesoderm induction during early embryogenesis as well as hematopoiesis. We studied the regulation of activin A, a homodimer of betaA-subunits, mRNA and protein in K562 erythroleukemia cells, which are known to be induced toward the erythroid lineage in response to activin or TGF-beta or toward the megakaryocytic lineage by the phorbol ester protein kinase C activator 12-O-tetradecanoylphorbol-13-acetate (TPA). Here we show by Northern blot analysis as well as by Western and ligand blotting that TPA strongly promotes activin betaA-subunit mRNA and activin A protein expression in K562 cells in time- and concentration dependent manner. In contrast, neither activin A nor TGF-beta induced betaA-subunit mRNA expression during erythroid differentiation in K562 cells. Interestingly, whereas activin type II receptors are not regulated during K562 cell differentiation (Hilden et al. (1994) Blood 83, 2163-2170), we now show that the activin type I and IB receptor mRNAs are clearly induced by TPA but not by activin or TGF-beta. We also show that the inducing effect of TPA on expression of activin betaA-subunit mRNA is potentiated by the protein kinase A activator 8-bromo-cAMP. We conclude that activin A and its type I receptors appear to be co-ordinately up-regulated during megakaryocytic differentiation of K562 cells. PMID- 10459862 TI - Unbound rather than total concentration and saturation rather than unsaturation determine the potency of fatty acids on insulin secretion. AB - Isolated mouse islets were used to compare the effects of three saturated (myristate, palmitate and stearate) and three unsaturated (oleate, linoleate and linolenate) long-chain fatty acids on insulin secretion. By varying the concentrations of fatty acid (250-1250 micromol/l) and albumin simultaneously or independently, we also investigated whether the insulinotropic effect is determined by the unbound or total concentration of the fatty acids. Only palmitate and stearate slightly increased basal insulin secretion (3 mmol/l glucose). All tested fatty acids potentiated glucose-induced insulin secretion (10-15 mmol/l), and the following rank order of potency was obtained when they were compared at the same total concentrations: palmitate approximately = stearate > myristate > or = oleate > or = linoleate approximately = linolenate. The effect of a given fatty acid varied with the fatty acid to albumin molar ratio, in a way which indicated that the unbound fraction is the important one for the stimulation of beta cells. When the potentiation of insulin secretion was expressed as a function of the unbound concentrations, the following rank order emerged: palmitate > myristate > stearate approximately = oleate > linoleate approximately = linolenate. In conclusion, the acute and direct effects of long chain fatty acids on insulin secretion are due to their unbound fraction. They are observed only at fatty acid/albumin ratios higher than those normally occurring in plasma. Saturated fatty acids are stronger insulin secretagogues than unsaturated fatty acids. Unbound palmitate is by far the most potent of the six common long-chain fatty acids. PMID- 10459864 TI - Natural variants of the beta isoform of the human glucocorticoid receptor do not alter sensitivity to glucocorticoids. AB - The beta isoform of the human glucocorticoid receptor, hGRbeta, is a product of alternative splicing of the hGR gene. The physiological function of this isoform is unknown up to now. Recent data are contradictory in that they either favor or argue against a role of hGRbeta as a repressor of the functional hGRalpha isoform. In the present study hGRbeta did not inhibit transcriptional activation of the MMTV-driven luciferase reporter gene by dexamethasone-activated hGRalpha in COS-1 cells. In addition, two naturally occurring variants of the hGRbeta isoform associated with altered sensitivity to glucocorticoids, termed hGRbeta R23K and hGRbeta-N363S, did not repress hGRalpha, even when overexpressed 10 fold. We conclude that the hGRbeta isoform, as well as two of its natural variants, do not act as dominant negative inhibitors of hGRalpha function and that the beta isoform does not appear to play a role in the regulation of glucocorticoid sensitivity. PMID- 10459863 TI - GLUT2 and glucokinase expression is coordinately regulated by sulfonylurea. AB - In the present study we examined the effect of sulfonylurea on the expression of the glucose transporter GLUT2 and the glucose phosphorylating enzyme Glucokinase (GK) in betaTC6-F7 cells; furthermore, we studied the modifications induced by sulfonylurea on glucose-responsiveness and -sensitivity. Results demonstrate that sulfonylurea increases GLUT2 and GK mRNA expression after 24 h in a dose dependent manner. On the contrary, after 48 and 72 h a time-dependent reduction of both GLUT2 and GK mRNA occurs. GLUT2 and GK protein expression follow the same modifications. Therefore, GLUT2 and GK are coordinately regulated by sulfonylurea, probably by a common mechanism. Glucose-induced insulin release is increased by sulfonylurea as well as glucose sensitivity. Our study suggests that short-term effect of sulfonylurea increases while long-term effect reduces the expression of glucose sensing elements. The long-term inhibitory effect on glucose sensing elements would explain the reduced insulin secretion occurring after chronic sulfonylurea treatment. PMID- 10459865 TI - Cloning of zebrafish activin type IIB receptor (ActRIIB) cDNA and mRNA expression of ActRIIB in embryos and adult tissues. AB - A full-length cDNA encoding for activin type IIB receptor (ActRIIB) was cloned from zebrafish embryos. It encodes a protein with 509 amino acids consisting of a signal peptide, an extracellular ligand binding domain, a single transmembrane region, and an intracellular kinase domain with predicted serine/threonine specificity. The extracellular domain shows 74-91% sequence identity to human, bovine, mouse, rat, chicken, Xenopus and goldfish activin type IIB receptors, while the transmembrane region and the kinase domain show 67-78% and 82-88% identity to these known activin IIB receptors, respectively. In adult zebrafish, ActRIIB mRNA was detected by RT-PCR in the gonads, as well as in non-reproductive tissues, including the brain, heart and muscle. In situ hybridization on ovarian sections further localized ActRIIB mRNA to cytoplasm of oocytes at different stages of development. Using whole-mount in situ hybridization, ActRIIB mRNA was found to be expressed at all stages of embryogenesis examined, including the sphere, shield, tail bud, and 6-7 somite. These results provide the first evidence that ActRIIB mRNA is widely distributed in fish embryonic and adult tissues. Cloning of zebrafish ActRIIB demonstrates that this receptor is highly conserved during vertebrate evolution and provides a basis for further studies on the role of activin in reproduction and development in lower vertebrates. PMID- 10459866 TI - Molecular mechanism for cooperation between Sp1 and steroidogenic factor-1 (SF-1) to regulate bovine CYP11A gene expression. AB - Bovine cholesterol side-chain cleavage cytochrome P450 (P450scc; product of the CYP11A gene) gene expression is regulated by gonadotropins via cAMP in the ovary, and by ACTH via cAMP in adrenal cortical cells. Previously, we characterized response elements located at -57/-32 and at -111/-101 bp in the 5'-flanking region of the bovine CYP11A gene required for cAMP-stimulated transcription in both mouse Y-1 adrenal tumor cells and bovine ovarian cells in primary culture, which bind SF-1 (or Ad4-BP) and Sp1, respectively. The role of these transcription factors in CYP11A transcription was further confirmed by deletion and mutation analyses. In addition, results obtained employing a double mutation of the Sp1- and SF-1-binding sites and a mammalian two-hybrid system indicate that Sp1 and SF-1 function cooperatively in the transactivation of the bovine CYP11A promoter in both bovine luteal cells and Y-1 cells. Here we report that SF 1 and Sp1 are able to associate with one another in vitro and in vivo. The NH2 terminal region of SF-1, especially the DNA-binding domain, is the binding site for Sp1. In addition, as CBP is a common coactivator required for the transcriptional activity of numerous transcription factors including nuclear receptors, we investigated whether CBP functions as a cofactor for the regulation of bovine CYP11A promoter activity. We show here that CBP enhanced the PKA induced CYP11A promoter activity, while a double mutation of both Sp1 and SF-1 sites within the CYP11A promoter region abolished CBP-induced activity. Furthermore, CBP stimulated Sp1-dependent transactivation, and a CBP/Sp1 complex in vivo was demonstrated by a co-immunoprecipitation assay. Also, CBP potentiated the transcriptional activity of GAL4-SF-1 in the presence of PKA. Thus, the cooperation between SF-1 and Sp1, required for the regulation of bovine CYP11A gene expression, is mediated by a direct protein-protein interaction and/or the common coactivator CBP. PMID- 10459867 TI - How to assess sympathetic activity in humans. AB - Sympathetic factors play a central role not only in cardiovascular homeostatic control but also in the pathogenesis and/or in the progression of several cardiovascular diseases, such as essential hypertension, myocardial infarction, cardiac arrhythmias and congestive heart failure. This explains why assessment of adrenergic neural function in humans has been, and certainly still remains, one of the major fields in cardiovascular research. The present paper will review in detail the haemodynamic, pharmacological, biochemical, neurophysiological, neurochemical and neural imaging techniques by which sympathetic activity is assessed in humans, highlighting the main advantages and limitations of each of them. Although plasma noradrenaline measurement represents a useful guide to assess sympathetic neural function, direct recording of sympathetic nerve traffic via microneurography and noradrenaline radiotracer methods have in recent years largely supplanted the plasma noradrenaline approach. This is because they allow (1) discrimination between the central or peripheral nature of increased plasma noradrenaline levels, and (2) precise estimation of the behaviour of regional sympathetic neural function both under physiological and pathological conditions. In contrast, the approach based on spectral analysis of heart rate and blood pressure signals has been shown to have important limitations which prevent the method from faithfully reflecting sympathetic cardiovascular drive. Neural imaging techniques, which require expensive technical support, allow direct visualization of sympathetic enervation of human organs, thus providing information on the 'in vivo' metabolism of noradrenaline in different cardiovascular districts. Although technical improvements have allowed a more precise assessment of human adrenergic function, no technique so far available can be viewed as a 'gold standard' with which the others might be compared. Limitations and disadvantages of the various techniques may be reduced if these methods are seen as being complementary and employed in combination, allowing more reliable information to be achieved on the sympathetic abnormalities characterizing cardiovascular diseases, and thus hopefully providing a stronger rationale for newer therapeutic approaches involving pharmacological modification of the sympathetic nervous system and adrenoreceptors. PMID- 10459868 TI - Guidelines for arterial hypertension: the echocardiography controversy. AB - While the recent evolution of guidelines for arterial hypertension also includes the need to evaluate coexistent cardiovascular risk factors and target organ damage in the base work-up for arterial hypertension, it does not include echocardiography systematically, because there is no evidence that information on LV geometry and function can modify management strategy in every circumstance, and there is concern about the technical variability of repeated echocardiographic examinations in the individual patient. The present issue of the Journal publishes a paper showing that adherence to the 1993 World Health Organization - International Society of Hypertension recommendations leaves untreated a proportion of patients with 'mild hypertension' who instead would have been treated if decision was also based on echocardiographic information on LV geometry. These findings challenge the most recent positions of national Societies, reserving the indication for echocardiography to patients with high risk (the vast majority). The present study appears indeed to reinforce the notion that echocardiographic examination might be very important in patients in whom, based on guidelines adherence, no pharmacological treatment would be required, whereas, based on the present evidence, echocardiographic information might be less important and perhaps superfluous for decision making in patients assigned to a high risk score, for whom aggressive treatment has been already scheduled. PMID- 10459869 TI - Hypertensive men who exercise regularly have lower rate of cardiovascular mortality. AB - BACKGROUND: Regular exercise has been associated in prospective studies with reduced incidence of cardiovascular disease (CVD) and death. OBJECTIVE: To assess in a cohort study whether there is a similar protective effect of regular exercise among hypertensive individuals. DESIGN: Population-based prospective cohort study. Spare time physical activity was assessed by structured interview. SETTING: Malmo, Sweden. PARTICIPANTS: Healthy men (n = 642) born in 1914. A baseline examination took place in 1969-1970. MAIN OUTCOME MEASURES: All-cause and cardiovascular mortality rates during 25 years of follow-up in relation to blood pressure and other risk factors for atherosclerosis. RESULTS: One-hundred (16%) men reported vigorous spare time physical activity. In this group, 31 had hypertension (blood pressure >160/95 mm Hg or treatment for hypertension), 47 were smokers and 39 had hyperlipidaemia. Among the 173 men with hypertension, vigorous physical activity was associated with markedly reduced rates of all cause (17.3 versus 40.0 deaths per 1000 person-years) and cardiovascular mortality (6.3 versus 21.0 deaths per 1000 person-years). The risk reductions associated with exercise remained statistically significant after adjustment for smoking, systolic blood pressure and antihypertensive therapy. The relative risk was 0.43 (confidence interval 0.22-0.82) for total mortality and 0.33 (confidence interval 0.11 -0.94) for CVD mortality. CONCLUSION: People who regularly perform physical activity constitute a heterogeneous group with regard to their exposure to known cardiovascular risk factors. Our results support the view that regular physical activity is associated with a reduced incidence of cardiovascular disease and death and suggest that this protective effect may be enhanced among hypertensive individuals. PMID- 10459870 TI - Carotid intima-media thickness and coronary heart disease risk factors in a low risk population. AB - OBJECTIVE: Coronary heart disease (CHD) risk factors have been consistently related to an increase in carotid intima-media thickness (IMT) in selected populations. However, few studies were population-based and furthermore little attention has been given to the influence of CHD risk factors on IMT in low-risk populations for CHD. DESIGN: We examined the association between carotid IMT and CHD risk factors in a large (n = 1013) and representative sample of middle-aged men and women in one of the European populations with the lowest CHD risk. METHODS: High-resolution B-mode ultrasonography of the common carotid arteries was performed. RESULTS: Age, smoking (not significant in women), body mass index, waist to hip ratio, systolic (SBP) and diastolic blood pressure, alcohol consumption, total and low-density lipoprotein cholesterol, triglycerides, glycaemia, fibrinogen (not significant in women), haematocrit (not significant in men) and insulin (not significant in women) were positively and significantly associated with mean IMT. High-density lipoprotein (HDL) cholesterol (not significant in women) was negatively and significantly associated with mean IMT. In a subsample of 355 men, IMT was not associated with angiotensin I-converting enzyme gene polymorphism. Multivariate analyses showed, in men, independent associations between mean IMT (0.61+/-0.11 mm) and age, pack-years, SBP, HDL cholesterol, alcohol and the interaction between age and alcohol. In women, only age and SBP were independently associated with mean IMT (0.58+/-0.09 mm). CONCLUSIONS: We found thinner IMT than those found in high-risk populations, suggesting that an increased IMT might reflect local atherosclerosis. Protective factors such as HDL cholesterol or regular and moderate alcohol consumption are probably important determinants of the early stages of atherosclerosis in these low-risk populations. PMID- 10459871 TI - Effects of a traditional lifestyle on the cardiovascular risk profile: the Amondava population of the Brazilian Amazon. Comparison with matched African, Italian and Polish populations. AB - OBJECTIVE: To determine the relationships between lifestyle and cardiovascular risk factors among the Brazilian Amondava, one of the world's most isolated populations. DESIGN: Cross-sectional, population-based study. Four age- and sex matched samples from Brazil Africa, Italy and Poland, representing different levels of modernization, were compared. Body weight, height, blood pressure, serum cholesterol and glycaemia were measured, and a standard questionnaire administered. Data concerning dietary habits and physical activity were collected. A personal socio-economic score was calculated, on the basis of type of economy, level of formal education, type of occupation, type of habitat, availability of piped water and electricity, main source of income, housing conditions, availability of radio, television or personal computer, knowledge of a second language, and organized health facilities. SETTING: Primary epidemiological screening, at an institution. RESULTS: Among the Amondava blood pressure was always <140/90 mm Hg, it did not increase with age and was not correlated with any other variable; 46.6% of subjects had systolic blood pressure <100 mm Hg. Blood pressure among the Amondava (109.6+/-11.1/69.5+/-6.4 mm Hg) was on average lower (P<0.0001) than in all other samples. Among the Amondava, the concentration of total cholesterol was always <200 mg/dl, i.e. similar to that of Africans whose diet included large amounts of vegetable foodstuffs; 90% had glycaemia (<80 mg/dl), and their mean value was the lowest (55.1+/-14.9 mg/dl) of all the groups. CONCLUSIONS: In addition to a possible genetic predisposition not analysed in this study, a traditional lifestyle (no contact with civilization, diet based on complex carbohydrates and vegetables, high energy expenditure) may protect against the development of hypertension, hypercholesterolaemia, and diabetes. PMID- 10459872 TI - Angiotensinogen gene and essential hypertension in the Japanese: extensive association study and meta-analysis on six reported studies. AB - BACKGROUND: Accumulating evidence has supported the pathophysiological role of angiotensinogen in essential hypertension. However, some studies of molecular genetics have implicated that there may be an ethnic variation concerning the disease susceptibility of the AGT gene. OBJECTIVES AND METHODS: To evaluate the importance of this candidate gene for hypertension, we undertook an extensive association study in the Japanese. This case-control study was conducted in a total of 1232 individuals consecutively enrolled in a single institution, divided into two subgroups: one subgroup comprised 254 hypertensive and 224 normotensive subjects and the other comprised 463 hypertensive and 291 normotensive subjects. A meta-analysis was subsequently performed on six Japanese studies including the present study. RESULTS: No significant association was observed between a molecular variant of AGT, Thr235, and hypertension status in our case-control study. Moreover, this finding was extendible to another AGT polymorphism, G-6A, one of the potential functional polymorphisms in the promoter region, because these two polymorphisms proved to be in complete linkage disequilibrium in the studied population. The meta-analysis revealed that the pooled estimate of the odds ratio across the studies was 1.22 (95% CI 1.05-1.42), and that there was significant evidence against homogeneity of the odds ratios among the studies included (phi2 = 19.8, df = 5, P = 0.0014). In particular, a large range of variation (60-83%) was found for the allele frequency of Thr235 among control subjects of the six Japanese case-control studies. CONCLUSIONS: Although the meta analysis appears in favour of association between the AGT variant and essential hypertension in the Japanese, there is considerable heterogeneity among the studies and the evidence is also rather borderline. Further comprehensive approaches are needed to resolve this debatable issue. PMID- 10459873 TI - Angiotensin-converting enzyme insertion-deletion polymorphism in normotensive and pre-eclamptic pregnancies. AB - OBJECTIVE: To investigate the hypothesis that pre-eclampsia is associated with a common insertion-deletion polymorphism in the angiotensin-converting enzyme gene. DESIGN: Seventy-two women with pre-eclampsia and 83 normotensive pregnant women participated in the study. Pre-eclampsia was defined as a blood pressure exceeding 140/90 mm Hg in a previously normotensive woman, associated with proteinuria in excess of 300 mg/l in a 24 h collection. Samples for fetal genotyping were available from 66 pregnancies complicated by pre-eclampsia and 79 normotensive pregnancies. METHODS: Maternal and fetal samples were genotyped at the insertion-deletion (I-D) polymorphism in intron 16 of the angiotensin converting enzyme gene by the polymerase chain reaction followed by agarose electrophoresis. RESULTS: Neither the I-D genotype distributions nor the allele frequencies differed significantly between pre-eclamptic and normotensive pregnancies in maternal or fetal samples (phi2 <0.3, not significant). The odds ratio for pre-eclampsia in women with the DD genotype, compared with the ID and II genotype, was 1.09 (95% confidence interval 0.55-2.16). The odds ratio associated with the DD genotype in the fetus was 1.14 (0.56-2.32). CONCLUSION: This study has found no evidence that the insertion-deletion polymorphism in the angiotensin-converting enzyme gene is associated with pre-eclampsia. PMID- 10459874 TI - Effects of chronic treatment with simvastatin on endothelial dysfunction in spontaneously hypertensive rats. AB - OBJECTIVE: To investigate the effects of chronic treatment with simvastatin (SV) on endothelium-dependent relaxation and ouabain-induced contractions in aortic rings from spontaneously hypertensive rats (SHR), comparing with normotensive Wistar-Kyoto rats (WKY). METHODS: After a 12-week period of administration of 1 or 2 mg/kg SV to SHR and WKY, systolic blood pressure (SBP) and vascular reactivity in endothelium-intact aortic rings were assessed. RESULTS: Relaxation in response to acetylcholine (ACh) in WKY remained unaltered, but in SHR treated with 1 mg/kg SV, enhanced ACh-induced relaxation (P<0.05 versus untreated SHR) reached values observed in untreated WKY. The 2 mg/kg treatment also improved ACh relaxation (P<0.01 and P<0.05 versus untreated SHR and WKY respectively). Inhibiting cyclo-oxygenase (COX) with indomethacin (INDO) improved ACh relaxation in SHR (P<0.05) but not in WKY, independent of treatment with SV. Inhibition of nitric oxide synthase (NOS) with N(G)-nitro-L-arginine (L-NOARG) abolished ACh relaxations in all cases (P<0.001). The result was unaltered when combining INDO plus L-NOARG. SV treatment also decreased ouabain-induced contractions in endothelium-intact aortic rings from SHR, diminishing the percentage effect of contraction from 64.56+/-2.95 (untreated SHR) to 26.98+/-7.06 and 38.10+/-8.21 (1 and 2 mg/kg treated SHR respectively). Response to ouabain in WKY was not significantly affected by SV treatment CONCLUSIONS: Chronic treatment of SHR with SV improves endothelium-dependent ACh relaxation of the aortic rings, probably by an NO-involving mechanism more than by inhibiting contractile COX-derived factors. An improvement in endothelial modulation of ouabain-induced contractions was also observed after treatment with SV in SHR, which might be due to an inhibition of a calcium-sodium exchanger. PMID- 10459875 TI - Reduced intestinal epithelial cell brush border membrane calcium transport in spontaneously hypertensive rats. AB - OBJECTIVE: The present study aimed to determine whether there were alterations in intestinal calcium homeostasis in the spontaneously hypertensive rats (SHR) and to identify at which interface of the intestinal epithelial cell (brush border or basolateral) this occurs. DESIGN: Controversy exists as to whether intestinal calcium transport is altered in association with hypertension. Studies using perfused duodenal segments of the SHR have shed little light on the problem; other studies have only measured calcium transport in brush border membrane vesicles. This study allows specific focus on calcium transport mechanisms at both the brush border and basolateral membrane using simultaneously prepared membrane vesicles. METHODS: Calcium transport was studied by measuring radiolabelled calcium (45Ca) uptake in isolated brush border and basolateral membrane vesicles, prepared from the small intestines of SHR and Wistar-Kyoto (WKY) rats. Calcium uptake was measured when vesicles were incubated in solutions containing different concentrations of ATP and calcium. Orientation and membrane marker assays were used to confirm the phenotypes of the two membrane vesicle preparations. RESULTS: ATP-dependent calcium efflux was only observed in the basolateral membrane, which contains the Ca2+ -ATPase pump. SHR brush border membrane vesicles displayed no significant increase in calcium incorporation, whereas WKY brush border vesicles showed a 500% increase in uptake (ANOVA, P<0.05, n = 7). CONCLUSIONS: This study indicates that deficiencies exist in SHR intestinal calcium transport at the brush border membrane of intestinal epithelial cells. While further studies are required to ascertain the exact mechanisms involved, postulated deficiencies in the actions of calcium regulating hormones at this membrane suggest the need for concurrent intake of a calcitrophic agent to assist calcium uptake at the brush border membrane. PMID- 10459876 TI - Membrane microviscosity, blood pressure and cytosolic pH in Dahl rats: the influence of plasma lipids. AB - OBJECTIVE: To determine the relationships between blood pressure, membrane microviscosity, plasma lipids and cytosolic pH in Dahl rats susceptible or resistant to salt hypertension. DESIGN AND METHODS: Blood pressure, plasma triglycerides and total cholesterol, platelet cytosolic pH (pHi) and the microviscosity of both outer membrane leaflet (TMA-DPH fluorescence anisotropy) and membrane lipid core (DPH fluorescence anisotropy) were studied in platelets and erythrocyte ghosts of Dahl salt-sensitive (SS/Jr) and salt-resistant (SR/Jr) rats fed either a low-salt diet (0.3% NaCl) until the age of 9, 15 or 24 weeks or a high-salt diet (4% NaCl) for 5 or 10 weeks after weaning. RESULTS: At low salt intake, DPH but not TMA-DPH anisotropy increased with age in platelets of SS/Jr rats. Chronic high salt intake was accompanied by an increase of DPH anisotropy in platelets but not in erythrocyte ghosts of SS/Jr rats. Platelet DPH anisotropy correlated positively with blood pressure of salt-loaded SS/Jr rats. Chronic high salt intake also reduced pHi in platelets, the regulation of which seemed to be related to the changes in TMA-DPH anisotropy. This especially concerns the thrombin-induced pHi rise which was inversely related to basal pHi, plasma lipids and TMA-DPH anisotropy. Altered membrane lipid composition might be the underlying mechanism because both membrane microviscosity and platelet pHi regulation were reported to correlate significantly with plasma triglycerides and/or cholesterol. CONCLUSIONS: Platelets of salt hypertensive Dahl rats are characterized by an increased microviscosity of membrane lipid core which correlated positively with blood pressure. The major influence of plasma triglycerides on DPH anisotropy should be taken into consideration when investigating the links between membrane microviscosity and blood pressure. On the other hand, the changes in microviscosity of the outer membrane leaflet might be involved in pHi regulation (probably through control of the Na+/H+ exchanger). PMID- 10459877 TI - Differential distribution of apolipoprotein E in young and aged spontaneously hypertensive and stroke-prone rats. AB - OBJECTIVES: To evaluate whether the differential distribution of apolipoprotein E among lipoprotein fractions and hepatic expression of the apolipoprotein E gene are causal factors in the regulation of lipid metabolism and physiological functions in young and aged spontaneously hypertensive and stroke-prone rats. DESIGN AND METHODS: Biochemical analyses were performed on serum and hepatic specimens from young (2-month-old) and aged (8-month-old) spontaneously hypertensive rats, stroke-prone spontaneously hypertensive rats and normotensive Wistar-Kyoto rats. Levels of apolipoprotein E among various lipoprotein fractions were determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Liver concentrations of apolipoprotein E mRNA were analyzed by Northern blotting and relative levels of apolipoprotein E mRNA in different strains of rats were compared. Statistical analysis was performed by measuring correlations between hepatic apolipoprotein E mRNA levels and biological parameters. RESULTS: Levels of apolipoprotein E in high-density and low-density lipoproteins were significantly lower in hypertensive rats than in age-matched normotensive Wistar- Kyoto rats. Although there was a significant increase in high-density lipoprotein apolipoprotein E contents in all aged animals, the elevation in aged hypertensive rats was much less than that in aged normotensive rats. Levels of apolipoprotein E in the very-low-density lipoprotein fraction were diminished in young stroke prone rats but were remarkably high in aged rats. Steady-state levels of apolipoprotein E mRNA increased with age in all strains of rats, whereas aged hypertensive rats exhibited lower apolipoprotein E mRNA levels than aged normotensive rats. CONCLUSIONS: The distribution of apolipoprotein E among various lipoprotein fractions was dramatically altered with age, and the alteration varied among different strains of rats. The differential distribution of apolipoprotein E in young and aged spontaneously hypertensive and stroke-prone rats suggests that apolipoprotein E could be a causal factor that disturbs the homeostasis of lipids and lipoproteins and perturbs physiological functions in hypertensive rats. PMID- 10459878 TI - Similar baroreflex bradycardic actions of atrial natriuretic peptide and B and C types of natriuretic peptides in conscious rats. AB - OBJECTIVE: We have previously shown that atrial natriuretic peptide (ANP) modulates cardiac barosensitive afferent pathways to enhance reflex bradycardia in rats. The present study examined whether B-type natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) also modulate heart rate reflex function. DESIGN: Baroreflex bradycardia was evoked by rapid (over 4-6 s) intravenous (i.v.) infusions of methoxamine (100 microg/kg; 'ramp' baroreflex technique) in the presence of infused i.v. natriuretic peptide and of vehicle (0.9% saline, 270 microl/h) in conscious adult Munich-Wistar rats. Initially a dose-response study to ANP (infused at 25, 50 and 100 pmol/kg per min i.v.) was performed in 10 rats to determine an appropriate dose for subsequent experiments with the other peptides. In a separate group of 11 animals, rat BNP-32 and rat CNP-22 were infused at 50 pmol/kg per min i.v. RESULTS: Reflex responses to ANP were dose related, with a significant increase in baroreflex sensitivity of 50+/-15% at the 25 pmol dose, 102+/-10% at the 50 pmol dose and 117+/-11% at 100 pmol dose (all P<0.05). BNP and CNP (50 pmol/kg/min i.v.) substantially increased baroreflex bradycardia (by 115+/-17% and 62+/-15%, respectively; P<0.05) compared to vehicle infusion. CONCLUSIONS: Both BNP and CNP augmented baroreflex slowing of heart rate in response to rapid increases in blood pressure in rats. Whereas other reports have shown marked differences in cardiovascular responses between the natriuretic peptides, particularly with CNP, our findings demonstrate an important common action of ANP, BNP and CNP to facilitate vagal heart rate baroreflexes. PMID- 10459879 TI - A heart-specific increase in cardiotrophin-1 gene expression precedes the establishment of ventricular hypertrophy in genetically hypertensive rats. AB - OBJECTIVE: Cardiotrophin-1 is a cytokine, a novel member of the interleukin-6 superfamily, which is isolated from mouse embryoid bodies. It is known to bind a gp130/ leukemia inhibitory factor (LIF) receptor heterodimer and to induce myocyte hypertrophy. Accumulating evidence indicates that a gp130 signaling pathway is involved in cardiac development and ventricular hypertrophy. METHODS: In order to elucidate the pathophysiologic significance of cardiotrophin-1 in ventricular hypertrophy associated with hypertension, we examined the level of cardiotrophin-1 mRNA in the ventricle of spontaneously hypertensive rats/Izm stroke-prone (SHRSP/Izm) in neonates, and at 4-, 12- and 20-weeks of age by Northern blot analysis. We also examined the gene expression of LIF by Northern blot and reverse transcription-polymerase chain reaction analyses. RESULTS: No significant difference was observed in the level of cardiotrophin-1 mRNA in the ventricle between SHRSP/ Izm and Wistar-Kyoto/Izm (WKY/Izm) neonates. However, the level of cardiotrophin-1 mRNA in the ventricle was significantly augmented in 4-week-old SHRSP/Izm, which did not yet show overt ventricular hypertrophy, and its augmented expression lasted for the duration of the experimental period. The difference in the level of cardiotrophin-1 mRNA between the two strains was most prominent at the age of 4 weeks. This augmented expression of the cardiotrophin-1 gene was not related to the severity of left ventricular hypertrophy. The level of cardiotrophin-1 mRNA in other organs, including the kidney and lung, showed no significant change with aging and was not different between the two strains. After long-term treatment with lisinopril, levels of cardiotrophin-1 mRNA were not changed, although it morphologically prevented the development of left ventricular hypertrophy. LIF mRNA was not detected in any ventricles examined by Northern blot analysis. CONCLUSIONS: The present study demonstrates that the expression of cardiotrophin-1 mRNA is increased in the early stage of ventricular hypertrophy in SHRSP/Izm and it remains elevated after hypertrophy has been established. However, it is unlikely that cardiotrophin-1 plays a mechanistic role in the development and maintenance of left ventricular hypertrophy in SHRSP/Izm. The present study also suggests that cardiotrophin-1, but not LIF, is a possible candidate for natural ligand of a gp130 signaling pathway in the heart. PMID- 10459880 TI - Can echocardiography identify mildly hypertensive patients at high risk, left untreated based on current guidelines? AB - OBJECTIVE: To determine whether the decision to treat uncomplicated mild hypertension with drugs, in accordance with the World Health Organization - International Society of Hypertension (WHO/ISH) guidelines based on a series of blood pressure (BP) measurements over 6 months, resulted in the treatment of patients at high risk on the basis of echocardiography. METHODS: One hundred and eighteen patients with mild hypertension (diastolic blood pressure 90-105 mm Hg and/or systolic blood pressure 140-180 mm Hg) were examined by echocardiography at inclusion and followed up for 6 months by a single physician unaware of the echographic results. RESULTS: Drug treatment was given to 48 patients, and 70 remained untreated. Treated patients had higher echographic indices than untreated patients (all P<0.05): left ventricular (LV) mass/body surface area (83.0+/-15.6 versus 75.3+/-14.8 g/m2), inter-ventricular septal thickness (9.7+/ 1.7 versus 8.5+/-1.3 mm), LV posterior wall thickness (8.4+/-1.1 versus 7.8+/-1.1 mm), relative wall thickness (0.37+/-0.06 versus 0.34+/-0.06). LV geometry was normal in 98 patients, and 20 had LV concentric remodelling. The 10-year coronary disease risk (Framingham equation) was higher in the 20 patients with concentric remodelling than in those with normal LV geometry (10.4 versus 4.2%; P<0.005). Nine of these 20 patients were still untreated at the end of the 6-month follow up period. CONCLUSION: Rigorous application of the WHO/ISH clinical guidelines in a group of mild hypertensive patients led to the treatment of patients with slightly higher LV mass and more concentric LV geometry than were found in those not treated. However, a high-risk subgroup, with concentric remodelling, was not identified and left untreated. PMID- 10459881 TI - Hyper-responsiveness to angiotensin II is related to cardiac structural adaptation in hypertensive subjects. AB - BACKGROUND: Angiotensin II has been found to be a growth stimulating factor for myocardial cells. In humans, angiotensin II infusion causes vasoconstriction in systemic and renal vasculature and leads to aldosterone secretion. Our hypothesis was that hyper-responsiveness to angiotensin II is related to left ventricular mass in human essential hypertension. METHODS AND RESULTS: In 30 normotensive individuals and 30 subjects with mild essential hypertension (white men, mean age 26+/-3 years), the responsiveness to angiotensin II was assessed by measuring changes in mean arterial pressure, renal blood flow, glomerular filtration rate and aldosterone secretion in response to i.v. angiotensin II infusion (0.5 and 3.0 ng/kg per min). The provoked changes to angiotensin II infusion were similar in the normotensive and hypertensive group with the exception of an exaggerated increase in mean arterial pressure in hypertensives (14+/-5 versus 10+/-5 mm Hg, P<0.001 at 3.0 ng/kg per min angiotensin II). The increase in mean arterial pressure was correlated with left ventricular mass in hypertensive subjects (angiotensin II 0.5 ng/kg per min: r = 0.49, P<0.005; angiotensin II 3.0 ng/kg per min: r = 0.35, P<0.05); no such correlation was found in the normotensive group. After taking into account baseline mean arterial pressure and body mass index, the increase in mean arterial pressure to angiotensin II 0.5 ng/kg per min was still correlated with left ventricular mass (partial r = 0.50, P<0.01). Similarly, the change of glomerular filtration rate but not of renal blood flow in response to angiotensin II 0.5 ng/kg per min was correlated with left ventricular mass, (r = 0.42, P<0.02) in the hypertensive group but not in the normotensive one. This relationship remained significant even after taking baseline glomerular filtration rate, mean arterial pressure and body mass index into account (partial r = 0.43, P<0.05). CONCLUSION: Hyper-responsiveness to angiotensin II is related to an increased left ventricular mass in hypertensive subjects independent of blood pressure. PMID- 10459882 TI - Blood pressure control in a hypertension hospital clinic. AB - OBJECTIVES: First, to evaluate the prevalence of clinic blood pressure (BP) control (BP < or = 140/90 mm Hg) in a representative sample of treated hypertensive patients followed in our hypertension clinic. Second, to assess in a subgroup of these patients: (a) the proportion of BP control with both clinic blood pressure (CBP < or =140/90 mm Hg) and daytime ambulatory blood pressure (ABP) (< or =132/85 mm Hg) criteria, and (b) the prevalence of echocardiographic left ventricular hypertrophy (LVH) (left ventricular mass index, LVMI>125 g/m2 in men and >110 g/m2 in women). DESIGN AND METHODS: Seven hundred consecutive hypertensive patients who attended our hypertension centre clinic during a period of 6 months and who had regularly been followed up by the same medical team were included in the study. BP was taken in the clinic by a doctor using a mercury sphygmomanometer with the participants seated. Seventy-four patients with similar demographic and clinical characteristics to the entire population of participants underwent complete echocardiographic examination and 24 h ABP monitoring. RESULTS: During follow-up, 352 of the treated patients had clinic BP < or =140/90 mm Hg, 198< or =160/95 mm Hg and 150>160/95 mm Hg, indicating that BP control was satisfactory in 50.3%, borderline in 28.3% and unsatisfactory in 21.4% of the cases. In the subgroup of 74 patients, the proportion of individuals with satisfactory clinic BP control (CBP< or =140/90 mm Hg) was higher (50.0 versus 33.6%) than with satisfactory ABP control (daytime ABP values < or =132/85 mm Hg). LVH was found in 21 of the 74 patients (28.3%): 12 of them had unsatisfactory CBP control and 19 had unsatisfactory ABP control. LVMI did not correlate with CBP values but only with ABP values (mean 24 h systolic r = 0.47, diastolic r = 0.40, P<0.001; mean daytime systolic r = 0.45, mean daytime diastolic r = 0.39, P<0.001; mean night-time systolic r = 0.38, mean night-time diastolic r = 0.38, P<0.001). CONCLUSION: This study demonstrates that hypertensive patients managed in a hypertension centre clinic have satisfactory CBP control in 50% of cases, but this rate seems to over-estimate the effective BP control during daily life. A large fraction of patients show persistence of LVH and this evidence of organ damage almost entirely concerns individuals with poor ABP control. PMID- 10459883 TI - Effects of chronic long-term therapy with calcium antagonists on cytogenetic damage in humans. AB - OBJECTIVE: To assess whether chronic long-term calcium antagonist therapy may increase genotoxicity, the chromosome aberration test, a widely accepted genotoxic assay, was used ex vivo in peripheral human lymphocytes of patients with or without long-term exposure to calcium antagonist therapy. METHODS AND RESULTS: In a case-control study design, we evaluated 30 ischaemic and/or hypertensive patients (22 males, eight females; age 59.4+/-1.5 years), under chronic calcium antagonist treatment (group I), for more than 3 years (4.4+/-0.34 years) and 30 age-matched subjects, without any previous exposure to calcium antagonists (group II). Venous blood samples were collected from the patients and cultures were set up for cytogenetic analysis by standard methods. For each subject, 100 metaphases were scored. The two groups showed similar values (mean +/- SEM) for percentage aberrant cells (group I 2.6+/-0.3 versus group II 2.5+/ 0.3, not significant), percentage structural aberrations (group I 1.9+/-0.3 versus group II 1.8+/-0.2, not significant) and percentage numerical aberrations (group I 0.70+/-0.2 versus group II 0.73+/-0.2, not significant). CONCLUSIONS: Long-term calcium antagonist therapy is not associated with an increased incidence of chromosomal indices of genotoxic damage in humans. PMID- 10459884 TI - The impact of stroke on informal carers: a literature review. AB - Stroke incidence is set to rise in Western societies as population projections predict an increase in the proportion of older people. Most of these stroke survivors are supported by close kin members who play an important role in the rehabilitation and care of this group. The objective of this review was to establish the following: (a) to evaluate the impact of the stroke on the informal carers' quality of life, (b) to identify factors which help carers to cope with their caring role, (c) to evaluate health service provision for stroke carers. A systematic literature search using BIDS-EMBASE, MEDLINE and PSYCHLIT, identified 31 relevant studies. These showed that most studies concentrated on carers' psychological health and the negative impact that the stroke had. Carers ability to cope with the stroke was enhanced both by the use of positive coping strategies and more concrete measures e.g. more stroke information. Furthermore, whilst most carers were generally satisfied with health services, the few interventions directed at improving carer outcomes showed mixed results. The studies reviewed had many limitations; few gave definitions of 'informal' carer and there was a predominant use of cross-sectional studies and non-standardised outcome measures. Future studies should broaden their research question to evaluate quality of life, using standardised measures to do this and employing either a longitudinal or randomised control design to improve the robustness of results. More studies are also needed evaluating the effectiveness of health services on carers' quality of life. PMID- 10459885 TI - 'Total pain', disciplinary power and the body in the work of Cicely Saunders, 1958-1967. AB - Pain first emerged as an area of clinical specialisation in the 1950s, but more recently has attracted wider interest from social scientists and clinicians who seek to expand its understanding to incorporate ideas about meaning, embodiment and culture. So far there have been few empirical studies which focus on how ideas and practices about pain are changing in modern healthcare. This paper addresses these issues through a specific case study of the early writings of Cicely Saunders in the period 1958-1967. A professional training in the three disciplines of nursing, social work and medicine, coupled with a strong personal religious faith, provided the biographical context for the development of Cicely Saunders' concern with pain. Through these influences we find in her work with dying patients an emphasis on pain as a key which unlocks other problems and as something which requires multiple interventions for its resolution. From here the concept of 'total pain' is formulated, to include physical, psychological, social, emotional and spiritual elements. This concept, which proved so important to the development of hospice clinical practice, is shown to have paradoxical and conflicting implications. Adopting current ideas about the social theory of the body, 'total pain' may be formulated either as a nomenclature of inscription, or as a nomenclature of facilitation. It is suggested that both of these may be at work in the discourse of 'total pain' and that an appreciation of each enhances our understanding of the concept. PMID- 10459886 TI - Determinants of depression and HIV-related worry among HIV-positive women who have recently given birth, Bangkok, Thailand. AB - HIV-infected pregnant women have been the focus of considerable research related to biomedical issues of mother-to-child transmission worldwide. However, there have been few reports on the psychological well-being of new mothers with HIV, either in developed or developing countries. As part of a perinatal HIV transmission and family impact study in Bangkok, predictors of psychological scales were evaluated from interview data (N = 129) collected 18-24 months postpartum. Standardised questionnaires were used to assess depressive symptoms and HIV-related worry. Depressive symptomatology and HIV-related worry were common amongst these women. Multivariate logistic regression analysis identified several factors that predicted these psychological outcomes. High depression scores were associated with women who were no longer in a relationship with their partner (odds ratio (OR) 5.72, confidence interval (CI) 2.18-14.97) and who used venting coping strategies (OR 2.15, CI 1.44-3.21). Higher levels of HIV-related worry were associated with women whose babies were HIV-infected (OR 3.51, CI 1.28 10.69), who had not disclosed their HIV status to others (OR 3.05, CI 1.29-7.24) and who reported that their HIV-infection was something about which their family would be ashamed (OR 3.44, CI 1.34-9.77). Based on the current findings, intervention strategies we propose are psychological interventions which address disclosure issues, feelings of shame and coping strategies as well as financial assistance for single mothers. Interventions that require few resources such as group counselling or support merit special consideration. PMID- 10459887 TI - Socio-demographic and spatial aspects of male mortality from HIV-AIDS related diseases in New South Wales, Australia, 1990-1994. AB - During 1990-1994, a very high proportion of males aged under 40 who died from HIV AIDs related diseases in New South Wales were never married. However, a significant minority of men aged 40-64 who died had been married at some stage in their lives and in the cities of Newcastle, Wollongong and in higher income areas of Sydney this proportion approached 40%. AIDS related mortality was over represented in professional, managerial, paraprofessional and service occupations even though educational levels were higher in these groups and, thus, presumably knowledge about risk behaviour and preventative behaviour. AIDS related mortality was also elevated among men with no stated occupation. Thus, with males, AIDS was in part a disease of affluence, even though the highest proportions of those dying resided in lower income areas, where marginalised persons may also be at risk. The proportion of deaths to men over age 40 was markedly higher than that in Australia in the 1980s. Elevated mortality with the New Zealand and the Americas birthplace groups may reflect overseas travel and exposure to risk in overseas countries, of persons domiciled in Australia. There was a very strong spatial pattern of AIDS-related mortality in the inner and eastern suburbs in Sydney and there is limited evidence of persons having migrated back to places of origin to be cared for by families. The impact on total mortality under age 65 in these localised areas was considerable. Prevention strategies should include the sensitive targeting of bisexual men, men generally having sex with men and those who are both gay and members of the drug sub-culture. PMID- 10459888 TI - The cost-effectiveness of introducing a varicella vaccine to the New Zealand immunisation schedule. AB - This study examined the cost-effectiveness of adding a varicella vaccine to an existing childhood immunisation schedule relative to a counterfactual where the varicella vaccine is available on a user-pays basis (the current New Zealand situation). The costs and consequences of chickenpox in an annual cohort of 57,200, 15-month old children were simulated for a 30-year period. The cohort simulation design captures the 'phasing-in' effects of routine varicella vaccination on the population. From a health care payer's perspective (medical costs only) every dollar invested in a vaccination programme would return NZ $0.67. However, from a societal point of view (which includes the value of work loss), a vaccination programme would return NZ $2.79 for every dollar invested. To implement a varicella vaccination programme covering 80% of 15-month old children in New Zealand would add more than NZ $1 million in net direct (health care) costs each year. However, the indirect cost savings from reduced losses of work-time exceed NZ $2 million annually. The net average health care cost per child vaccinated over the 30-year modelling period was $54 whereas the cost savings from work-loss averted averaged $101 per child vaccinated. Total cost savings to society of $47 per child vaccinated, on average, could be gained from a vaccination programme. The finding that the addition to vaccination costs resulting from a routine programme (including the cost of complications from the vaccine) were greater than the offsetting health care cost savings from reduced incidence of chickenpox were robust to a sensitivity analysis on all assumptions within plausible ranges. Overall cost-effectiveness estimates were most sensitive to assumptions regarding lost work-time, the discount rate, and the price and efficacy of the vaccine. Estimates were relatively insensitive to changes in assumptions regarding health care utilisation. PMID- 10459889 TI - Physical activity social support and middle- and older-aged minority women: results from a US survey. AB - BACKGROUND: Many American adults remain sedentary despite many known health benefits. Research on the determinants of physical activity have indicated that social support is one of the strongest correlates, but little is known about this relation in important subgroups of middle and older-aged women. OBJECTIVE: The purpose of this study was to assess the association of physical activity-related social support on several measures of physical activity in a national sample of minority women. A unique aspect of these measures is the inclusion of vigorous household tasks and occupational physical activities. METHODS: The US Women's Determinants Study was conducted in 1996-1997. The survey was a modified-random sample, telephone survey of 2912 Black, Hispanic, American Indian/Alaskan Native, and White women age 40 and older. A composite score of physical activity social support (PASS) was analyzed as the independent variable in logistic regression analyses. Four measures of physical activity levels served as the dependent variables. A separate analysis was done to distinguish PASS from friends versus PASS from relatives. The potential confounding effect of race/ethnicity, marital status, age, income and education were evaluated and adjusted in the models. RESULTS: Hispanic women were more likely to have high PASS scores than the other racial/ethnic groups. Odds ratios indicate that subjects with high levels of PASS were significantly less likely to be sedentary than those with low support, even after adjusting for race/ethnicity. While there were significant associations among levels of social support and physical activity, this was not true for the measure of "regular exercise." There was no significant difference between the contribution of "friend" support versus "family" support on all four measures on physical activity. DISCUSSION: Based on our results, enhancing social support may be an important aspect of interventions aimed at increasing physical activity in a population of sedentary women of various racial/ethnic backgrounds. Also, "regular exercisers" in this population appear to be less reliant social support to maintain their behavior. PMID- 10459890 TI - The effect on morbidity of variability in deprivation and population stability in England and Wales: an investigation at small-area level. AB - We seek to determine whether variability in deprivation at small area level, and population stability, influence standardised morbidity ratios in England and Wales. A regression analysis was conducted with data from the 1991 British Census, in order to explain variation in morbidity. Both an area deprivation score (for electoral wards) and the within-area variability of deprivation scores were examined as possible determinants of morbidity (self-reported, limiting, long-term illness). Particular attention was focused on a spatially-sensitive measure of the variability of deprivation scores within a wider 'locality'. There was a significant, positive relationship between age-standardised limiting, long term illness and deprivation. The variation in area deprivation scores within the small areas themselves was also significant and positive. However, the variation in deprivation scores calculated for both an electoral ward and its contiguous neighbours (the locality) was slightly more significant. Areas with higher relative levels of in-migration also had significantly lower standardised morbidity ratios. Multivariate models showed that the deprivation score, the variation in deprivation scores for the broader locality, and the measure of migration, were all significant in combination. Residual analysis showed that many areas in London had lower levels of morbidity than expected, while electoral wards in the coal mining valleys of South Wales had higher levels than expected. We conclude that, for small areas (wards) in England and Wales, morbidity is related to deprivation, variation in deprivation within and surrounding each area, and the proportion of the population that are migrants. Variations in deprivation influence standardised morbidity rates, and policies which widen inequalities will influence health outcomes. Resource allocation based simply on measures of deprivation, which ignore population change within the area and variations in deprivation in the locality, may be inefficient. PMID- 10459891 TI - Sexuality education in Russia: defining pleasure and danger for a fledgling democratic society. AB - Public health indicators have plummeted throughout Russia since the collapse of the Soviet Union, with exponential increases in sexually transmitted diseases reported among this society's young adults. Newly developing sexuality education programs provide insights into the ways local health providers interpret such public health challenges and conceptualize the educational needs of Russian youth. Moreover, these initiatives reveal the impact of both Soviet-era discourses and more recent, international anti-abortion activism on contemporary thinking about sexual health matters. This article explores the implicit and sometimes explicit ways that sex education lectures are being driven by debates over the significance of the Soviet past and anxieties over the perceived chaos of current transformations. Drawing on material from lectures, fieldwork, and interviews with sex educators, I argue that sexuality education efforts reveal a persistent ambivalence between the hope to promote individual autonomy from state interests and the presumed need to control sexual expression and reproductive practices within an emerging moral economy of post-Soviet Russia. PMID- 10459892 TI - Different tuberculosis in men and women: beliefs from focus groups in Vietnam. AB - After decades in decline, tuberculosis (TB) has been increasing worldwide. In 1993, the World Health Organisation declared TB a global emergency. Passive case finding is an important part of TB control programmes, and this is strongly affected by people's perceptions and beliefs of TB and society's behaviour towards TB sufferers. The aim of this study was to describe the perceptions and beliefs of Vietnamese people regarding TB and its risk factors with special reference to differences between men and women. Sixteen focus group discussions (FGDs) were organised in four districts representing different regions in Vietnam and consisting of men and women, TB patients and non-TB participants. In general, participants had good knowledge of TB being a dangerous, contagious and infectious disease, caused by germs. However, traditional beliefs in different types of TB still exist, mainly among older people in rural areas, but also resorted to by other people once ill. Four main types of TB were reported: (1) 'Lao truyen' (hereditary TB), handed down from older generations to latter ones through 'family blood', regardless of sexes; (2) 'Lao luc' (physical TB), caused by hard work, more men affected; (3) 'Lao tam' (mental TB), caused by too much worrying-more women affected; and (4) 'Lao phoi' (lung TB), dangerous and caused by TB germs, transmitted through the respiratory system-more men affected. Other general risk factors were also mentioned. Men were perceived to get TB more often than women, as they were more exposed to risk factors during both work and leisure time. These traditional beliefs may contribute to long delays to TB diagnosis and increased social stigma and isolation of TB patients and their families due to erroneous beliefs in transmission routes. Our findings demonstrate areas where TB control programmes may be improved. PMID- 10459893 TI - What price information? Modelling threshold probabilities of fetal loss. AB - This paper is an extension of previous work in which an alternative method of measuring the benefits of antenatal screening was proposed. The method is based on the elicitation of threshold probabilities of fetal loss at which women would be indifferent between having and not having an amniocentesis for the prenatal diagnosis of fetal abnormalities. The aim of this paper is to extend the previous work by modelling the preferences of a larger sample of women and investigating the consistency and validity of their responses. The threshold probabilities are elicited using standard gambles and modelled using Tobit estimation. The results indicate that it is possible to model these probabilities in this way and that it is possible to obtain a high degree of consistency in response to standard gamble questions. While establishing the validity of the responses is more difficult there is some evidence that the technique can provide valid responses. PMID- 10459894 TI - Understanding social variation in cardiovascular risk factors in women and men: the advantage of theoretically based measures. AB - Many studies have attempted to understand observed social variations in cardiovascular disease in terms of sets of intermediate or confounding risk factors. Tests of these models have tended to produce inconsistent evidence. This paper examines the relationships to cardiovascular risk factors or two theoretically based measures of social position. It shows that the strength of the relationships between social position and cardiovascular risk factors varies according to the definition of social position which is used: there is a closer relationship between most health behaviours and the Cambridge scale, an indicator of 'general social advantage and lifestyle', whereas the Erikson-Goldthorpe schema, which is based on employment relations and conditions, is more strongly related to work control and breathlessness. The implications of these findings for understanding the conflicting evidence in other studies of health inequalities are then discussed. The paper concludes that inconsistencies between studies may be in part due to unexamined differences between the conceptual bases of the measures of social position they use, combined with a failure to make explicit the hypothetical mechanisms of effect. If neither the conceptual basis of the measure of social position, nor the links between social position and health outcome tested in each study are clear, inconsistencies between studies will be difficult to interpret, making policy recommendations highly problematic. PMID- 10459895 TI - Tragedy of variant Creutzfeldt-Jakob disease. PMID- 10459896 TI - Is intensive drug therapy appropriate for older patients? PMID- 10459897 TI - Uncloaking the meningococcus: dynamics of carriage and disease. PMID- 10459898 TI - Time to reconsider treatment options for intracranial aneurysms. PMID- 10459899 TI - Trials stopped early: too good to be true? PMID- 10459900 TI - Cough and gastro-oesophageal reflux. PMID- 10459901 TI - The real millennium bug. PMID- 10459902 TI - Cationic lipid-mediated CFTR gene transfer to the lungs and nose of patients with cystic fibrosis: a double-blind placebo-controlled trial. AB - BACKGROUND: We and others have previously reported significant changes in chloride transport after cationic-lipid-mediated transfer of the cystic fibrosis transmembrane conductance regulator (CFTR) gene to the nasal epithelium of patients with cystic fibrosis. We studied the safety and efficacy of this gene transfer to the lungs and nose of patients with cystic fibrosis in a double-blind placebo-controlled trial. METHODS: Eight patients with cystic fibrosis were randomly assigned DNA-lipid complex (active) by nebulisation into the lungs followed 1 week later by administration to the nose. Eight control patients followed the same protocol but with the lipid alone (placebo). Safety was assessed clinically, by radiography, by pulmonary function, by induced sputum, and by histological analysis. Efficacy was assessed by analysis of vector specific CFTR DNA and mRNA, in-vivo potential difference, epifluorescence assay of chloride efflux, and bacterial adherence. FINDINGS: Seven of the eight patients receiving the active complex reported mild influenza-like symptoms that resolved within 36 h. Six of eight patients in both the active and placebo groups reported mild airway symptoms over a period of 12 h following pulmonary administration. No specific treatment was required for either event. Pulmonary administration resulted in a significant (p<0.05) degree of correction of the chloride abnormality in the patients receiving active treatment but not in those on placebo when assessed by in-vivo potential difference and chloride efflux. Bacterial adherence was also reduced. We detected no alterations in the sodium transport abnormality. A similar pattern occurred following nasal administration. INTERPRETATION: Cationic-lipid-mediated CFTR gene transfer can significantly influence the underlying chloride defect in the lungs of patients with cystic fibrosis. PMID- 10459903 TI - How should age affect management of acute myocardial infarction? A prospective cohort study. AB - BACKGROUND: About 75% of patients with acute myocardial infarction are older than 70 years, but patients in this age group are commonly treated less vigorously than younger patients. This differential treatment may partly reflect clinicians' misconceptions about the outlook of such patients, and the importance of age in clinical decisions. We examined how age does and should affect the management of patients and risk stratification in acute myocardial infarction. METHODS: In this prospective cohort study, we recruited 1225 consecutive patients admitted with acute myocardial infarction to a district general hospital in east London. The primary endpoint was death. We used tabulation and regression methods to analyse the association between age group and clinical variables. FINDINGS: Patients aged 70 years or older took a longer time to arrive in hospital and were less likely to receive thrombolysis or discharge beta-blockers than patients younger than 60 years: odds ratio 0.63 (95% CI 9.45-0.88) for thrombolysis and 0.25 (0.16-0.37) for beta-blockade, adjusted for sex, diabetes, previous acute myocardial infarction, Q wave infarction, and left-ventricular failure. Left-ventricular failure was the strongest independent predictor of death within 1 year of infarction with a hazard ratio of 4.76 (3.53-6.43), adjusted for age, sex, diabetes, and Q wave infarction. Patients aged 70 years or older without left ventricular failure had significantly better survival at 1 year after acute myocardial infarction than patients under 60 years with left-ventricular failure. 70.8% (62.2-78.2) of the older patients who survived to hospital discharge were still alive 3 years later. INTERPRETATION: Elderly patients with acute myocardial infarction were treated less vigorously than younger patients. The prognosis of acute myocardial infarction, however, was substantially affected by the development of left-ventricular failure and other clinical indices, such that many older patients had a better outlook than younger patients with adverse clinical factors. In planning risk-based management, consideration of age independently of clinical status is inappropriate. PMID- 10459904 TI - Early surgery compared with watchful waiting for glue ear and effect on language development in preschool children: a randomised trial. AB - BACKGROUND: Otitis media with effusion (OME) is the most common cause of hearing loss in children and is generally treated by elective surgery. We compared in children with persistent OME the effect on speech and language development of immediate surgery (ventilation-tube insertion) and watchful waiting before surgery. METHODS: We did a randomised controlled trial with masked outcome assessment in Bristol Children's Hospital, Bristol, UK. We included 186 children born between April 1, 1991, and Dec 31, 1992, who had confirmed bilateral OME and bilateral hearing impairment of 25-70 dB of at least 3 months' duration. Children were randomly assigned surgery within 6 weeks (n=92), or 9 months of watchful waiting (n=90), after which bilateral tube insertion was done if required. We assessed hearing loss, expressive language, and verbal comprehension at 9 months and 18 months. FINDINGS: At 9 months, standardised scores for expressive language and verbal comprehension differed between groups with marginal significance after adjustment for baseline differences (p=0.04 and p=0.028, respectively). At 9 months, verbal comprehension and expressive language skills in the watchful waiting group were 3.24 months behind those in the early-surgery group. The watchful-waiting group was delayed on these two measures compared with their age expected levels. 18 months after randomisation, 85% of children in the watchful waiting group had received surgery and groups did not differ significantly. INTERPRETATION: There is some benefit from ventilation-tube insertion for expressive language and verbal comprehension but the timing of surgery is not critical. PMID- 10459905 TI - Predictive value of gadolinium-enhanced magnetic resonance imaging for relapse rate and changes in disability or impairment in multiple sclerosis: a meta analysis. Gadolinium MRI Meta-analysis Group. AB - BACKGROUND: Reliable prognostic factors are lacking for multiple sclerosis (MS). Gadolinium enhancement in magnetic resonance imaging (MRI) of the brain detects with high sensitivity disturbance of the blood-brain barrier, an early event in the development of inflammatory lesions in MS. To investigate the prognostic value of gadolinium-enhanced MRI, we did a meta-analysis of longitudinal MRI studies. METHODS: From the members of MAGNIMS (European Magnetic Resonance Network in Multiple Sclerosis) and additional centres in the USA, we collected data from five natural-course studies and four placebo groups of clinical trials completed between 1992 and 1995. We included a total of 307 patients, 237 with relapsing disease course and 70 with secondary progressive disease course. We investigated by regression analysis the relation between initial count of gadolinium-enhancing lesions and subsequent worsening of disability or impairment as measured by the expanded disability status scale (EDSS) and relapse rate. FINDINGS: The relapse rate in the first year was predicted with moderate ability by the mean number of gadolinium-enhancing lesions in monthly scans during the first 6 months (relative risk per five lesions 1.13, p=0.023). The predictive value of the number of gadolinium-enhancing lesions in one baseline scan was less strong. The best predictor for relapse rate was the variation (SD) of lesion counts in the first six monthly scans which allowed an estimate of relapse in the first year (relative risk 1.2, p=0.020) and in the second year (risk ratio=1.59, p=0.010). Neither the initial scan nor monthly scans over six months were predictive of change in the EDSS in the subsequent 12 months or 24 months. The mean of gadolinium-enhancing-lesion counts in the first six monthly scans was weakly predictive of EDSS change after 1 year (odds ratio=1.34, p=0.082) and 2 years (odds ratio=1.65, p=0.049). INTERPRETATION: Although disturbance of the blood-brain barrier as shown by gadolinium enhancement in MRI is a predictor of the occurrence of relapses, it is not a strong predictor of the development of cumulative impairment or disability. This discrepancy supports the idea that variant pathogenetic mechanisms are operative in the occurrence of relapses and in the development of long-term disability in MS. PMID- 10459906 TI - First-line tuberculosis therapy and drug-resistant Mycobacterium tuberculosis in prisons. AB - BACKGROUND: We assessed a programme of tuberculosis control in a prison setting in Baku, Azerbaijan. The programme used first-line therapy and DOTS (directly observed treatment, short course). METHODS: 467 patients had sputum-positive tuberculosis. Their treatment regimens followed WHO guidelines, and they had regular clinical examinations and dietary supplements. Isolates were tested by standard methods for resistance to isoniazid, rifampicin, ethambutol, and streptomycin in three laboratories. Treatment success was defined as three consecutive negative sputum smears at end of treatment. Factors independently associated with treatment failure were estimated by logistic regression. FINDINGS: Drug-resistance data on admission were available for 131 patients. 55% of patients had strains of Mycobacterium tuberculosis resistant to two or more antibiotics. Mortality during treatment was 11%, and 13% of patients defaulted. Overall, treatment was successful in 54% of patients, and in 71% of those completing treatment. 104 patients completed a full treatment regimen and remained sputum-positive. Resistance to two or more antibiotics, a positive sputum result at the end of initial treatment, cavitary disease, and poor compliance were independently associated with treatment failure. INTERPRETATION: The effectiveness of a DOTS programme with first-line therapy fell short of the 85% target set by WHO. First-line therapy may not be sufficient in settings with a high degree of resistance to antibiotics. PMID- 10459907 TI - Expression of cytokeratin 20 redefines urothelial papillomas of the bladder. AB - BACKGROUND: Most non-invasive urothelial tumours of the bladder are diagnosed as papillary carcinomas in accordance with the WHO classification and because the identification of papillomas is difficult by routine histology; some patients are therefore misdiagnosed. This practice is associated with psychological morbidity for the patient and may also skew cancer statistics. Cytokeratin 20 (CK20) is a sensitive marker of urothelial differentiation. We investigated whether this marker could be used in the identification of urothelial papillomas and used the rate of recurrence as an indicator to assess the biological behaviour of such tumours. METHODS: In a prospective study, immunocytochemistry for CK20 was done on tumours of all patients who presented for the first time with non-invasive papillary bladder tumours. We classified the expression pattern of CK20 as normal or abnormal at the time of initial diagnosis. We recorded time to first biopsy proven recurrence or length of follow-up when no recurrence was observed. FINDINGS: Of 58 consecutive patients, ten had tumours with a normal pattern of CK20 expression. No patients developed further tumours during the follow-up (median 18 [range 13-28] months). By contrast, 30 (73%) of the 41 evaluable patients with tumours that showed abnormal CK20 expression developed further tumours; the median time to a second tumour was 6 (2-24) months. The only factor that had a significant effect on the outcome of patients in terms of recurrence was expression of CK20 (p<0.0001). INTERPRETATION: Normal urothelial differentiation, as evidenced by a normal pattern of CK20 expression, is retained in a proportion of non-invasive papillary urothelial tumours and thus justifies use of the term urothelial papilloma. A large-scale study is needed to investigate the outcome of patients with such tumours. PMID- 10459908 TI - A blistering rash and swollen knees. PMID- 10459909 TI - Deaths from variant Creutzfeldt-Jakob disease. PMID- 10459910 TI - QT lengthening and life-threatening arrhythmias associated with fexofenadine. PMID- 10459911 TI - Risperidone-induced tardive dystonia and psychosis. PMID- 10459912 TI - Blood vessels change in the mesencephalon of patients with Parkinson's disease. PMID- 10459913 TI - Platelet glycoprotein IIIa polymorphism, aspirin, and thrombin generation. PMID- 10459914 TI - Fulminant liver failure associated with flutamide therapy for hirsutism. PMID- 10459915 TI - Inhibition of proinflammatory cytokine production by pravastatin. PMID- 10459916 TI - Pulmonary disease severity in men with deltaF508 cystic fibrosis and residual chloride secretion. PMID- 10459917 TI - Ultrasonography of the small bowel after oral administration of anechoic contrast solution. PMID- 10459918 TI - Insects in the transmission of Mycobacterium ulcerans infection. PMID- 10459919 TI - UK "Bristol case" inquiry formally opened. PMID- 10459920 TI - New day dawns for research on circadian rhythms. PMID- 10459921 TI - Peace at all cost--the challenge of Sierra Leone. PMID- 10459922 TI - Paradox of the global emergency of tuberculosis. PMID- 10459923 TI - An African perspective on the threat of tuberculosis and HIV/AIDS--can despair be turned to hope? PMID- 10459924 TI - The threat of multidrug resistance: is tuberculosis ever untreatable or uncontrollable? PMID- 10459925 TI - Tuberculosis, Russia, and the Holy Grail. PMID- 10459926 TI - Sex, gender, and tuberculosis. PMID- 10459927 TI - Children and tuberculosis: protecting the next generation? PMID- 10459928 TI - Success with the DOTS strategy. PMID- 10459929 TI - Post DOTS, post genomics: the next century of tuberculosis control. PMID- 10459930 TI - A donor's perspective on tuberculosis in international health. PMID- 10459931 TI - Operating theatre design. PMID- 10459932 TI - Beta-blockers in heart failure. PMID- 10459933 TI - Beta-blockers in heart failure. PMID- 10459934 TI - Colorectal cancer. PMID- 10459935 TI - Colorectal cancer. PMID- 10459936 TI - Colorectal cancer. PMID- 10459937 TI - Primary aldosteronism in general practice. PMID- 10459938 TI - Primary aldosteronism in general practice. PMID- 10459939 TI - Iron-deficiency anaemia in premenopausal women. PMID- 10459940 TI - Iron-deficiency anaemia in premenopausal women. PMID- 10459941 TI - A deadly thorn: a case of imported melioidosis. PMID- 10459942 TI - A deadly thorn: a case of imported melioidosis. PMID- 10459943 TI - Achondroplasia associated with pelvic lipomatosis. PMID- 10459944 TI - Achondroplasia associated with pelvic lipomatosis. PMID- 10459945 TI - Mortality in the CHAOS trial. Cambridge Heart Antioxidant Study. PMID- 10459946 TI - Patients' concerns about clinical trials in Japan. PMID- 10459948 TI - Public health in Celtic Britain. PMID- 10459947 TI - Fly control and shigellosis. PMID- 10459949 TI - The Nobel chronicles. 1946: Hermann Joseph Muller (1890-1967). PMID- 10459950 TI - Below the fold. Eastbourne. PMID- 10459951 TI - Institutionalised racism in health care. PMID- 10459952 TI - Short-course zidovudine for prevention of perinatal infection. PMID- 10459953 TI - Proof that glucose-insulin-potassium provides metabolic protection of ischaemic myocardium? PMID- 10459954 TI - Prediction of burden of hip fracture. PMID- 10459955 TI - Prosthetic joint infections: controversies and clues. PMID- 10459956 TI - Academia and industry: lessons from the unfortunate events in Toronto. PMID- 10459957 TI - Short-course zidovudine for perinatal HIV-1 transmission in Bangkok, Thailand: a randomised controlled trial. Bangkok Collaborative Perinatal HIV Transmission Study Group. AB - BACKGROUND: Many developing countries have not implemented the AIDS Clinical Trials Group 076 zidovudine regimen for prevention of perinatal HIV-1 transmission because of its complexity and cost. We investigated the safety and efficacy of short-course oral zidovudine administered during late pregnancy and labour. METHODS: In a randomised, double-blind, placebo-controlled trial, HIV-1 infected pregnant women at two Bangkok hospitals were randomly assigned placebo or one zidovudine 300 mg tablet twice daily from 36 weeks' gestation and every 3 h from onset of labour until delivery. Mothers were given infant formula and asked not to breastfeed. The main endpoint was babies' HIV-1-infection status, tested with HIV-1-DNA PCR at birth, 2 months, and 6 months. We measured maternal plasma viral concentrations by RNA PCR. FINDINGS: Between May, 1996, and December, 1997, 397 women were randomised; 393 gave birth to 395 live-born babies. Median duration of antenatal treatment was 25 days, and median number of doses during labour was three. 99% of women took at least 90% of scheduled antenatal doses. Adverse events were similar in the study groups. Of 392 babies with at least one PCR test, 55 tested positive: 18 in the zidovudine group and 37 in the placebo group. The estimated transmission risks were 9.4% (95% CI 5.2 13.5) on zidovudine and 18.9% (13.2-24.2) on placebo (p=0.006; efficacy 50.1% [15.4-70.6]). Between enrolment and delivery, women in the zidovudine group had a mean decrease in viral load of 0.56 log. About 80% of the treatment effect was explained by lowered maternal viral concentrations at delivery. INTERPRETATION: A short course of twice-daily oral zidovudine was safe and well tolerated and, in the absence of breastfeeding, can lessen the risk for mother-to-child HIV-1 transmission by half. This regimen could prevent many HIV-1 infections during late pregnancy and labour in less-developed countries unable to implement the full 076 regimen. PMID- 10459958 TI - Short-course oral zidovudine for prevention of mother-to-child transmission of HIV-1 in Abidjan, Cote d'Ivoire: a randomised trial. AB - BACKGROUND: In Africa, the risk of mother-to-child transmission of HIV-1 infection is high. Short-course perinatal oral zidovudine might decrease the rate of transmission. We assessed the safety and efficacy of such a regimen among HIV 1-seropositive breastfeeding women in Abidjan, Cote d'Ivoire. METHODS: From April, 1996, to February, 1998, all consenting, eligible HIV-1-seropositive pregnant women attending a public antenatal clinic in Abidjan were enrolled at 36 weeks' gestation and randomly assigned placebo or zidovudine (300 mg tablets), one tablet twice daily until the onset of labour, one tablet at onset of labour, and one tablet every 3 h until delivery. We used HIV-1-DNA PCR to test the infection status of babies at birth, 4 weeks, and 3 months. We stopped the study on Feb 18, 1998, when efficacy results were available from a study in Bangkok, Thailand, in which the same regimen was used in a non-breastfeeding population. FINDINGS: 280 women were enrolled (140 in each group). The median duration of the prenatal drug regimen was 27 days (range 1-80) and the median duration of labour was 7.5 h. Treatment was well tolerated with no withdrawals because of adverse events. All babies were breastfed. Among babies with known infection status at age 3 months, 30 (26.1%) of 115 babies in the placebo group and 19 (16.5%) of 115 in the zidovudine group were identified as HIV-1 infected. The estimated risk of HIV-1 transmission in the placebo and zidovudine groups were 21.7% and 12.2% (p=0.05) at 4 weeks, and 24.9% and 15.7% (p=0.07) at 3 months. Efficacy was 44% (95% CI -1 to 69) at age 4 weeks and 37% (-5 to 63) at 3 months. INTERPRETATION: Short-course oral zidovudine was safe, well tolerated, and decreased mother-to child transmission of HIV-1 at age 3 months. Substantial efforts will be needed to ensure successful widespread implementation of such a regimen. PMID- 10459960 TI - Antihypertensive drugs in very old people: a subgroup meta-analysis of randomised controlled trials. INDANA Group. AB - BACKGROUND: Beneficial clinical effects of treatment with antihypertensive drugs have been shown in middle-aged patients and in those hypertensive patients over 60 years old, but whether treatment is beneficial in patients over 80 years old is not known. METHODS: We collected data from all participants aged 80 years and over in randomised controlled trials of antihypertensive drugs through direct contact with study investigators. Our primary outcome was fatal and non-fatal stroke. Secondary outcomes were death from all causes, cardiovascular death, fatal and non-fatal major coronary and cardiovascular events, and heart failure. FINDINGS: There were 57 strokes and 34 deaths among 874 actively treated patients, compared with 77 strokes and 28 stroke deaths among 796 controls, representing 1 non-fatal stroke prevented for about 100 patients treated each year. The meta-analysis of data from 1670 participants aged 80 years or older suggested that treatment prevented 34% (95% CI 8-52) of strokes. Rates of major cardiovascular events and heart failure were significantly decreased, by 22% and 39%, respectively. However, there was no treatment benefit for cardiovascular death, and a non-significant 6% (-5 to 18) relative excess of death from all causes. INTERPRETATIONS: The inconclusive findings for mortality contrast with the benefit of treatment for non-fatal events. Results of a large-scale specific trial are needed for definite conclusion that antihypertensive treatment is beneficial in very elderly hypertensive patients. Meanwhile, an age threshold beyond which hypertension should not be treated cannot be justified. PMID- 10459959 TI - 6-month efficacy, tolerance, and acceptability of a short regimen of oral zidovudine to reduce vertical transmission of HIV in breastfed children in Cote d'Ivoire and Burkina Faso: a double-blind placebo-controlled multicentre trial. DITRAME Study Group. DIminution de la Transmission Mere-Enfant. AB - BACKGROUND: Zidovudine reduces the rate of vertical transmission of HIV in non breastfed populations. We assessed the acceptability, tolerance, and 6-month efficacy of a short regimen of oral zidovudine in African populations practising breastfeeding. METHODS: A randomised double-blind placebo-controlled trial was carried out in public clinics of Abidjan, Cote d'Ivoire, and Bobo-Dioulasso, Burkina Faso. Eligible participants were women aged 18 years or older, who had confirmed HIV-1 infection and pregnancy of 36-38 weeks duration, and who gave written informed consent. Exclusion criteria were severe anaemia, neutropenia, abnormal liver function, and sickle-cell disease. Women were randomly assigned zidovudine (n=214; 300 mg twice daily until labour, 600 mg at beginning of labour, and 300 mg twice daily for 7 days post partum) or matching placebo (n=217). The primary outcome was the diagnosis of HIV-1 infection in the infant on the basis of sequential DNA PCR tests at days 1-8, 45, 90, and 180. We compared the probability of infection at a given age in the two groups. Analyses were by intention to treat. FINDINGS: Women were enrolled between September, 1995, and February, 1998, when enrolment to the placebo group was stopped. Analysis was based on 421 women and 400 lifeborn infants. Baseline demographic, clinical, and laboratory characteristics were similar in the two groups. The Kaplan-Meier probability of HIV infection in the infant at 6 months was 18.0% in the zidovudine group (n=192) and 27.5% in the placebo group (n=197; relative efficacy 0.38 [95% CI 0.05-0.60]; p=0.027). Adjustment for centre, period of recruitment, mode of delivery, maternal CD4-cell count, duration of labour, prolonged rupture of membranes, and duration of breastfeeding did not change the treatment effect. The proportions of women taking more than 80% of the planned maximum dose were 75% before delivery, 81% during labour, and 83% post partum, without statistical difference between the groups. No major adverse biological or clinical event was reported in excess among women and children of the zidovudine group. INTERPRETATION: A short course of oral zidovudine given during the peripartum period is well accepted and well tolerated, and provides a 38% reduction in early vertical transmission of HIV-1 infection despite breastfeeding. PMID- 10459962 TI - Hip fractures in Finland between 1970 and 1997 and predictions for the future. AB - BACKGROUND: Hip fractures in elderly people are common worldwide, and the predicted ageing of populations is increasing the burden of these fractures on health-care systems. However, prediction of the true increases in number of patients needing treatment requires exact knowledge of whether the number of hip fractures is rising more rapidly than can be accounted for by demographic changes alone. We aimed to make such a prediction for people aged 50 years or more in Finland. METHODS: All patients aged 50 years or more, who were admitted to hospitals in Finland during 1970-97 for primary treatment of first hip fracture were selected from the National Hospital Discharge Register. The age-specific and age-adjusted fracture incidences were expressed as the number of patients per 100,000 individuals per year, and prediction of the number, and incidence of hip fractures in Finland (population 5 million) until the year 2030 was calculated with a regression model. FINDINGS: The number of hip fractures in Finnish people aged 50 or more rose during the study period, from 1857 in 1970 to 7122 in 1997. The corresponding fracture incidence (per 100,000 people) increased from 163 to 438. The age-adjusted incidence of hip fractures also showed a steady increase from 1970 to 1997: in women, from 292 to 467, and in men, from 112 to 233. If this trend continues, the number of hip fractures in Finland will be almost three fold higher in the year 2030 than in 1997. INTERPRETATION: The number of hip fractures in elderly Finnish men and women is increasing at a rate that cannot be explained merely by demographic changes. The precise reasons for this are not known, but deterioration in age-adjusted bone-mineral density and strength, with accompanying increase in the age-adjusted incidence of injurious falls of the elderly, could partly account for the development. PMID- 10459961 TI - Adjuvant intra-arterial iodine-131-labelled lipiodol for resectable hepatocellular carcinoma: a prospective randomised trial. AB - BACKGROUND: Resection of hepatocellular carcinoma is potentially curative, but local recurrence is common. In this prospective randomised trial, we aimed to find out if one dose of postoperative adjuvant intra-arterial iodine-131-labelled lipiodol could reduce the rate of local recurrence and increase disease-free and overall survival. METHODS: Patients who underwent curative resection for hepatocellular carcinoma and recovered within 6 weeks were randomly assigned one 1850 MBq dose of 131I-lipiodol or no further treatment (controls). We compared rates of recurrence and disease-free and overall survival (the primary endpoints) between the two groups by intention to treat. We planned an interim analysis when 30 patients (both groups together) had been followed up for a median of 2 years, with the intention of stopping early if the between-group difference in disease free survival was significant (p=0.029). FINDINGS: Between April, 1992, and August, 1997, we recruited 43 patients: 21 received intra-arterial 131I-lipiodol and 22 received no adjuvant treatment. During a median follow-up of 34.6 (range 14.1-69.7) months, there were six (28.5%) recurrences among the 21 patients in the adjuvant treatment, compared with 13 (59%) in the controls (p=0.04). Median disease-free survival in the treatment and control groups was 57.2 (0.4-69.7) and 13.6 (2.1-68.3) months, respectively (p=0.037). 3-year overall survival in the treatment and control groups was 86.4% and 46.3%, respectively (p=0.039). The interim analysis showed a significant increase in disease-free survival in the treatment group compared with the controls (p=0.01), so we closed the trial early. 131I-lipiodol had no significant toxic effects. INTERPRETATION: In patients with hepatocellular carcinoma, one 1850 MBq dose of intra-arterial 131I lipiodol given after curative resection significantly decreases the rate of recurrence and increases disease-free and overall survival. PMID- 10459963 TI - A woman who left her wheelchair. PMID- 10459964 TI - Deprivation and hospital admission for infectious intestinal diseases. PMID- 10459965 TI - Monitoring antibody responses to cancer vaccination with a resonant mirror biosensor. PMID- 10459966 TI - Abciximab not RGD peptide inhibits von Willebrand factor-dependent platelet activation under shear. PMID- 10459967 TI - Reactivation of herpesvirus infections after vaccinations? PMID- 10459968 TI - Sexual dysfunction with protease inhibitors. PMID- 10459969 TI - Hypermenorrhea associated with ritonavir. PMID- 10459970 TI - N-3 fatty acids as a risk factor for haemorrhagic stroke. PMID- 10459971 TI - Codeine for pruritus in primary billiary cirrhosis. PMID- 10459972 TI - An iceberg of childhood coeliac disease in the Netherlands. PMID- 10459973 TI - ACE genotype and risk of high altitude pulmonary hypertension in Kyrghyz highlanders. PMID- 10459974 TI - Ageing eyes retain their mystery. PMID- 10459975 TI - When impotence leads contraception. PMID- 10459976 TI - South African budget targets AIDS campaigns. PMID- 10459977 TI - Peritoneal dialysis. AB - Peritoneal dialysis has now become an established form of renal replacement therapy; nearly half the patients on dialysis in the UK are treated in this way. Survival of patients is now equal to that with haemodialysis. However, long-term peritoneal dialysis (>8 years) is limited to a small percentage of patients because of dropout to haemodialysis for inherent complications of peritoneal dialysis--peritonitis, peritoneal access, inadequate dialysis, and patient related factors. However, improvements in the understanding of the pathophysiological processes involving the peritoneal membrane have paved the way for advances in the delivery of adequate dialysis, more biocompatible dialysis fluids, and automated peritoneal dialysis. Other technical advances have led to a reduction in peritonitis. Peritoneal dialysis is an important dialysis modality and should be used as an integral part of RRT programmes. PMID- 10459978 TI - Evidence and ethics. PMID- 10459979 TI - Science, ethics, and the future of research into maternal infant transmission of HIV-1. Perinatal HIV Intervention Research in Developing Countries Workshop participants. AB - Effective, feasible interventions to prevent perinatal transmission of HIV-1 in developing nations are an urgent necessity. Scientific issues of concern include a need to identify other effective antiretroviral agents; to define the shortest effective course of therapy; to assess interventions other than antiretroviral agents; and to investigate interventions that may reduce HIV-1 transmission via breastfeeding. Sound scientific design is fundamental to all research studies. Ethical standards must guide such studies and include the necessity that the problem studied be a health priority in the host country; that the highest standard of care attainable in the country be assured to participants; that the health-care resources of the country not be harmed; that the informed consent of participants be obtained; and that a process of discussion ensure that a successful intervention will be considered for implementation. There are circumstances in which a no-antiretroviral comparison may be ethically justified. PMID- 10459980 TI - Seizing the opportunity: collaborative initiatives to reduce HIV and maternal mortality. PMID- 10459981 TI - Possible interaction between sildenafil and HIV combination therapy. PMID- 10459982 TI - CD38/CD8 expression and HAART failure. PMID- 10459983 TI - Adherence to HAART in HIV. PMID- 10459984 TI - Unrecognised iron deficiency in critical illness. PMID- 10459985 TI - Unrecognised iron deficiency in critical illness. PMID- 10459986 TI - Unrecognised iron deficiency in critical illness. PMID- 10459987 TI - Unrecognised iron deficiency in critical illness. PMID- 10459988 TI - ACE inhibitors and prevention of aspiration pneumonia in elderly hypertensives. PMID- 10459989 TI - Vascular disruptive syndromes after exposure to misoprostol or chorionic villus sampling. PMID- 10459990 TI - Thrombotic vascular events after change of statin. PMID- 10459991 TI - Thrombotic vascular events after change of statin. PMID- 10459992 TI - Thrombotic vascular events after change of statin. PMID- 10459993 TI - Octreotide for nausea and vomiting after chemotherapy and stem-cell transplantation. PMID- 10459994 TI - NSAIDs and hepatic reactions. PMID- 10459995 TI - Affirmative action. PMID- 10459996 TI - Affirmative action. PMID- 10459997 TI - Developing country twinning programmes in paediatric oncology. PMID- 10459998 TI - Save the rain forest--it saved our skin. PMID- 10459999 TI - Web opens window on mental illness art. PMID- 10460000 TI - The Nobel chronicles. 1944: Joseph Erlanger (1874-1965); and Herbert Spencer Gasser (1888-1963). PMID- 10460001 TI - Blame the scholar, not the discipline. PMID- 10460002 TI - Cross-country comparisons suggest shared risk factors for carcinomas, including male lung adenocarcinoma and colon cancer development. AB - Smoking is clearly the major risk factor for both squamous cell carcinomas (SCCs) and adenocarcinomas (ACs) of the lung, although less so for the latter, where other influences appear to be important. In order to determine whether cross country comparisons might provide evidence of shared risk factors with cancers in other organs of males, an examination of IARC/WHO data for cancer incidence was made for countries/registries in Europe, North America, Australia and Asia. Significant simple correlations, which persisted on partial analysis, were observed between lung SCCs and tumors of the larynx (P < 0.001), but not the buccal cavity or oesophagus, along with a link to rectal ACs (P < 0.001). Incidences of lung ACs also correlated with those for colon ACs (P < 0.001) but not lung SCCs. Oesophageal ACs were only related to colon cancers at the simple correlation level, this not persisting on partial analysis or separation into Asian and Western groups. The results suggest that blood borne factors, like hormones, may be important as determinants for the increasingly prevalent lung AC. PMID- 10460003 TI - Cadherin and catenin expression in normal human bronchial epithelium and non small cell lung cancer. AB - Cadherins are transmembrane cell adhesion molecules (CAMS) that mediate cell-cell interactions and are important for maintenance of epithelial cell integrity. This function is dependent on an indirect interaction between the cytoplasmic domain of the cadherin molecule with three cytoplasmic proteins known as alpha-, beta-, and gamma-catenin (-cat). Growing evidence suggests that alterations in cadherin or catenin expression or function may be important to the development of an invasive or metastatic phenotype. Immunohistochemical techniques were used to study the expression of the two major epithelial cadherins, E-cadherin (E-cad) and P-cadherin (P-cad) as well as alpha- and gamma-cat in normal bronchial epithelium and in a series of carefully TMN-staged pulmonary adenocarcinomas (n = 21) and squamous cell carcinomas (n = 7). The cadherin profile of normal pseudostratified bronchial epithelium was heterogeneous. Basilar cells strongly expressed P-cad, alpha- and gamma-cat, while columnar cells moderately expressed E-cad, alpha- and gamma-cat. In contrast to other epithelial tumors, E-cad on non small cell lung carcinomas was actually upregulated, however, a decrease in P-cad expression was noted in 68%. At least one cadherin or catenin was downregulated, compared to normal bronchial epithelium, in 82% of tumors examined. With the exception of an association between loss of P-cad expression and poorly differentiated state, changes in cadherin and catenin expression levels were not significantly correlated to tumor stage, cell type, or nodal status. These findings illustrate that alteration of expression of cadherins and catenins are often found in non-small cell lung carcinoma when compared to the progenitor bronchial epithelium, and may play a role in the development of the malignant phenotype. PMID- 10460004 TI - Percutaneous brachytherapy for small-sized non-small cell lung cancer. AB - A patient with a small-sized pulmonary adenocarcinoma was successfully treated by percutaneous high dose rate interstitial brachytherapy alone. The patient, who had an adenocarcinoma with 12-mm diameter in the lingular lobe of left lung, was judged to be inoperable because of poor pulmonary function due to emphysema and extensive pleural adhesion. The tumor was punctured with a 21-gauge fine applicator needle followed by the introduction of an iridium 192 (192Ir) radioactive source through the applicator needle using a remote afterloader. The tumor was irradiated for 225.1 s in one fraction. The tumor was in the inside of the iso-dose line of 40 Gy. The delivered doses calculated at nine reference points, which were 12.5 mm distant from the center of the tumor, distributed between 19.225 and 32.169 Gy, with a mean of 24.8 Gy. No apparent side effect including pneumothorax and hemoptysis was observed. The tumor shrank and showed no increment of the size for about 2 years. PMID- 10460005 TI - Low-grade B-cell lymphoma of mucosa-associated lymphoid tissue in the lung: a report of a case with pleural dissemination. AB - A 79-year-old man with an abnormal shadow on a chest radiograph was referred to our hospital for further examination. Open lung biopsy revealed numerous nodules on visceral pleura and the tumor, obtained by wedge resection of the left upper lobe of the lung, consisted of centrocyte-like cells and lymphoplasmacytoid cells, expressing CD20 and CD79a. These cells invaded bronchiolar epithelium to form lymphoepithelial lesions. The pleural-based nodules were similarly composed of the same cells as those in the left upper lobe tumor. To our knowledge, pleural dissemination of low grade B-cell lymphoma of mucosa-associated lymphoid tissue has not been reported previously. PMID- 10460006 TI - Summary of the proceedings of the United States--Japan lung cancer clinical trials summit: San Francisco, CA, 20-22 November, 1998. PMID- 10460007 TI - Aromatase and sex steroid receptors in human vena cava. AB - Among sex steroids, especially estrogen metabolism has been considered to play a role in the function and pathology of human veins. We investigated the expression and activity of the estrogen-producing enzyme aromatase and estrogen receptor (ER) in human vena cava to assess possible in situ biosynthesis of estrogens and their modes of action. We first examined aromatase expression by immunohistochemistry in human inferior vena cava obtained from 29 autopsy cases (11 males, 18 females, 63.6 +/- 3.0 years old). We then semiquantitated the level of aromatase mRNA by reverse transcriptase-polymerase chain reaction in 24 cases and aromatase activity by 3H-water assay in 15 cases to examine whether or not and in which cell types aromatase was expressed. We also studied alternative use of multiple exon 1s of its gene and immunolocalization of 17beta-hydroxysteroid dehydrogenase type I (17beta-HSD I), which converts estrone produced by aromatase to estradiol, a biologically active estrogen and ER. Aromatase and 17beta-HSD I immunoreactivity were both detected in smooth muscle cells (SMC) of the media in all the cases and in endothelial cells (EC) in 20 and 22 cases, respectively. ER immunoreactivity was detected in SMC of vena cava in 21 cases. The amount of aromatase mRNA was significantly greater in the cases utilizing 1c (I.3) or 1d (P.II) of exon 1 (9 cases, 191.1 +/- 26.3 attomol/ng total RNA) than those utilizing 1b (I.4) as the promoter (14 cases, 50.6 +/- 13.0 attomol/ng total RNA) (p < 0.01). Significant correlation (p < 0.05) was observed between the amount of aromatase mRNA and aromatase activity in 15 cases examined. No significant correlation was detected between the amount of aromatase mRNA or aromatase labeling index and the ER status. These results suggest that estrone and estradiol are produced in the human vena cava and that their production is mediated by aromatase and 17beta-HSD I, respectively but not all of these locally synthesized estrogens may not work directly in situ. PMID- 10460008 TI - Effect of 22-oxa-1,25-dihydroxyvitamin D3 on human thyroid cancer cell growth. AB - To examine whether synthetic vitamin D3 analog, 22-oxa-1,25(OH)2D3 (OCT) has an inhibitory effect on the growth of thyroid carcinoma, we tested the in vitro and in vivo effects of OCT on the growth of a well-differentiated thyroid cancer cell line, NPA. OCT bound to its receptor at the same rate as 1,25(OH)2D3, and inhibited the proliferation of NPA cells in vitro in a dose-dependent manner, similar to that observed with 1,25 (OH)2D3. Northern blot analysis showed that steady-state and fetal bovine serum-stimulated levels of c-myc mRNA were suppressed after 0.5-4 hour treatment with OCT. Transfection studies with the deletion mutants of the 5'-up-stream flanking region of c-myc/chloramphenicol acetyltransferase chimera genes indicated the presence of an OCT responsive element between -410 and -106. Next, we examined OCT effects in implanted NPA tumor cells in nude mice. OCT showed no remarkable hypercalcemic effect compared to 1, 25 (OH2)D3, but OCT and 1, 25 (OH2)D3, had no significant inhibitory effect in vivo after either intra-tumor or intra-peritoneum injection. Our results demonstrate that OCT inhibits the proliferation of well-differentiated thyroid cancer in an in vitro system associated with the suppression of c-myc mRNA, but this inhibitory effect was not reproducible in in vivo model. PMID- 10460009 TI - Regulation of c-fos gene induction and mitogenic effect of transforming growth factor-beta1 in rat articular chondrocyte. AB - We have previously reported that type I transforming growth factor beta (TGF beta1) is a potent stimulator of cell growth in articular chondrocytes. In this study, we examined the mechanism of TGF-beta1 induced cellular proliferation by using cultured rat articular chondrocytes (CRAC). A time-course study of [3H]thymidine incorporation upon TGF-beta1 (1 ng/mL) or 10% fetal bovine serum stimulation revealed that TGF-beta1 directly stimulates DNA synthesis in CRAC. Pretreatment with H7, an inhibitor for protein kinase C (PKC), completely blocks TGF-beta1-induced proliferation. Since TGF-beta1 has been shown to transduce signals through MAP kinase cascades, we investigated the induction of several protooncogenes by Northern blotting. TGF-beta1 addition causes an immediate and transient induction of c-fos but not myc or jun mRNA. Furthermore, this c-fos expression is not inhibited by cycloheximide, but is completely abolished by pretreatment with TPA, so that the c-fos gene is a direct target of TGF-beta1 signalling and PKC is involved in this c-fos induction. To refine our understanding of TGF-beta1 regulation of the c-fos promoter region, we performed chloramphenicol acetyltransferase (CAT) assays. A serial deletion analysis of the c-fos promoter region reveals a TGF-beta1 responsive element in a region between 403 and -329 bp upstream of the transcription initiation site. We attempted gel shift assays on this response element with CRAC nuclear extracts. Although this region contains a sis-inducible binding element, we fail to detect specific DNA protein complexes. Our results, however, suggest that TGF-beta1 acts as a primary mitogen in CRAC and this mitogenic activity requires PKC activation. Finally, the subsequent induction of c-fos expression occurs through an as yet unidentified transactivation mechanism. PMID- 10460010 TI - MRI detection of suprasellar germinoma causing central diabetes insipidus. AB - This is a case report of an 18-year-old man with central diabetes incipidus (DI). An MRI done three months after the onset of the DI did not disclose a responsible lesion. Four months later, a second MRI showed the location of the tumor origin at the upper pituitary stalk and median eminence. Eight months later, the tumor occupied the hypothalamic area. The tumor became large and contrast-making enough to be visible on MRI between 3 and 4 months after the onst of DI. Besides the suprasellar tumor, another mass was noted in the pineal region. The growth pattern of the latter mass corresponded well to that of the former. Although the MRI is a sensitive diagnostic tool for the detection of intracranial tumors, no adequate rationale has been given as to how the MRI might be repeated for children and adolescents who have been diagnosed as having the central DI, when their initial MRIs may have been normal. In our patient, the superconductive thin slice MRI revealed the suprasellar germinoma 4 months after the onset. The suprasellar and pineal tumors in this report originated and developed simultaneously. This may indicate a multi-center origin of the tumor. Another possibility is a very early dissemination from the onset of the tumor development. PMID- 10460011 TI - Osmoregulation of vasopressin secretion in patients with the syndrome of inappropriate antidiuresis associated with central nervous system disorders. AB - To clarify the characteristics of vasopressin (AVP) secretion in patients with the syndrome of inappropriate antidiuresis (SIAD) related to central nervous system disorders, we examined the response of AVP secretion to osmotic stimulus by hypertonic saline infusion and analyzed the possible causative factors in six patients with SIAD associated with head trauma or cerebral infarction. Hyponatremia developed after head trauma in four patients and cerebral infarction in two patients. In all patients the clinical state and laboratory findings fulfilled the criteria for SIAD, which was supported by either nonsuppressible plasma AVP levels or effectiveness of treatments with water restriction, demeclocycline, nonpeptide V2 AVP antagonist or diphenylhydantoin. Although patterns of plasma AVP response to the osmotic stimulus varied, plasma AVP concentrations neither increased nor decreased to undetectable levels with a rise in plasma osmolality. In one patient, plasma AVP levels responded to increasing plasma osmolality when plasma osmolality normalized; in which the threshold and the sensitivity of osmostat were normal. In two other patients, AVP secretion responded to plasma osmolality after the treatment. The changes in AVP secretion were not due to nonosmotic stimuli for AVP release. In conclusion, this study shows that patients with SIAD and central nervous system disorders may have persistent AVP secretion with a loss of hypotonic suppression such as found in patients with adrenal insufficiency or depletional hyponatremia in central nervous system disorders, indicating that careful evaluation is necessary to determine the relationship between persistent AVP secretion and the pathogenesis of hyponatremic disorders. PMID- 10460012 TI - Polymorphism of homopolymeric glutamines in coactivators for nuclear hormone receptors. AB - Some of the recently identified coactivators which interact with members of nuclear hormone receptors contain a stretch of homopolymeric glutamines (poly-Q). Length of poly-Q in several genes are known to be polymorphic in healthy subjects, and extraordinary expansion of poly-Q in specific genes is known to cause neurodegenerative disorders. In the present study, we investigated whether such polymorphism can be observed in two coactivators, CBP (CREB [cyclic AMP responsive element binding protein]-binding protein) and AIB1/ACTR (amplified in breast cancer-1/ACTR, also called RAC3/TRAM-1). The genomic regions encoding the poly-Q were amplified by means of PCR using fluorescence labeled primer and analyzed by an automatic sequencer. While contiguous glutamine residues inAIB1/ACTR ranged from 26 to 32 with a heterozygosity of 54%, no polymorphism could be observed in poly-Q of CBP among 54 unrelated subjects. These results suggest that the residue in CBP may play a critical role in the function so that individuals with CBP containing different sizes of poly-Q might have been eliminated. It has been reported that AIB1/ACTR is overexpressed in some of the cell lines derived from breast cancer. If the length of poly-Q alters the stability of AIB1/ACTR and/or potency to enhance hormone action through nuclear receptors, the length of poly-Q is likely to be one of the genetic factors affecting not only susceptibility to breast cancers but also the sensitivity to hormones. This polymorphism should also be tested in patients with neurodegenerative disorders of unknown cause. PMID- 10460013 TI - A male patient presenting with major clinical symptoms of glucocorticoid deficiency and skeletal dysplasia, showing a steroid pattern compatible with 17alpha-hydroxylase/17,20-lyase deficiency, but without obvious CYP17 gene mutations. AB - We report the case of a 17-year-old boy with delayed puberty, who presented a complexity of clinical problems. An analysis of steroid hormones led to a diagnosis of 17alpha-hydroxylase/17,20-lyase deficiency (17OHD). Unlike typical cases of 17OHD, however, the patient had pubertal development without medical intervention. In addition, he never exhibited the symptoms of mineralocorticoid excess, showing instead the symptoms of glucocorticoid deficiency, including fatigability, emaciation, and weight-loss induced by minor infection. He also had dysmorphic features, which comprised marfanoid habitus, arachnodactyly and putative craniosynostosis. The combination of these malformations substantially resembled that of Shprintzen-Goldberg syndrome. Direct sequencing of the CYPl7 gene did not reveal any significant aberrations in the exons or exon-intron boundaries. We speculate that the association of partial combined 17OHD with the Shprintzen-Goldberg phenotype in the present patient may result from an aberration of a hitherto unknown gene that controls both steroid hormone synthesis and skeletal development. PMID- 10460014 TI - Changes in interleukin-1beta mRNA expression in the rat ovary during the estrous cycle in response to lipopolysaccharide. AB - The changes in interleukin-1 (IL-1) beta mRNA expression and the number of macrophages were studied in the ovary during the estrous cycle in rats and after intraperitoneal injection of lipopolysaccharide (LPS, 2 mg/body) 2 hours before autopsy. IL-1beta mRNA expression was very low in the ovary, and there was no statistically significant change during the estrous cycle. Hybridization signals of IL-1beta mRNA were localized intensely in the thecal layer, moderately in the corpora lutea, and slightly in granulosa cells of the ovary during the cycle. The number of macrophages seen mainly in the hilum and interstitium significantly increased on proestrus compared with other estrous days. LPS significantly increased IL-1beta mRNA expression on each day with the highest response to LPS at 1500 h on proestrus, and caused an increase in the number of macrophages in the ovary within 2 hours. These results indicate that IL-1beta mRNA expressions are low during the estrous cycle in rats, and proestrus is the day of maximal IL 1beta synthesis in response to LPS. The increase in IL-1beta synthesis caused by LPS might be due to at least the influx of macrophages into the ovary. PMID- 10460015 TI - Estrogen supplementation for bone dematuration in young epileptic man treated with anticonvulsant therapy; a case report. AB - We encountered a young man treated with anticonvulsant therapy who had greatly reduced bone mineral density. An 18-year-old man was admitted to our hospital for shoulder pain and further evaluation of decreased bone mineral density. He had been treated with anticonvulsants, including phenytoin, phenobarbital, valproic acid and zonisamide for seizures. Although testosterone was found within the normal range for adult men, the serum estrogen concentration was below the detection limit (< 10 pg/ml) and his wrist epiphyses were not yet closed. After 10 months of treatment with the conjugated estrogen, both his height and weight showed improvement, while his bone mineral density and bone age were increased. These findings suggested that estrogen therapy had a significant effect on his skeletal growth and bone maturation in man. This is the first report showing the beneficial effect of estrogen supplementation in an epileptic man receiving treatment with anticonvulsants. PMID- 10460016 TI - Inhibitory effect of thyrotropic hormone on apoptosis induced by actinomycin D in a functioning rat thyroid cell line. AB - Apoptosis appears to play important roles in physiological and pathological processes in the endocrine system including the thyroid, but little is known about the regulation of apoptosis in the thyroid. The functioning rat thyroid cell line (FRTL-5), a cloned cell line of differentiated thyroid cells, hardly undergoes apoptosis. In this study we examined the factors which prevent FRTL-5 cells from undergoing apoptosis. After culturing FRTL-5 cells in medium with and without TSH, actinomycin D (AMD) or cycloheximide (CHX) was added. CHX did not induce apoptosis. AMD induced apoptosis in FRTL-5 cells cultured in medium lacking TSH, as confirmed by the presence of DNA fragmentation, together with nuclear fragmentation and condensation, but AMD did not induce apoptosis in FRTL 5 cells cultured in the presence of TSH. Furthermore, the fact that AMD did not induce apoptosis in FRTL-5 cells cultured with dibutyryl cyclic AMP (Bt2cAMP), or forskolin suggests that TSH has an inhibitory effect on apoptosis in FRTL-5 cells via the TSH-cAMP pathway. PMID- 10460017 TI - Synergistic enhancement by 12-O-tetradecanoylphorbol-13-acetate and dibutyryl cAMP of 1alpha,25-dihydroxyvitamin D3 action in human promyelocytic leukemic HL 60 cells. AB - We have reported that dibutyryl cAMP (dbcAMP), an activator of cAMP-dependent protein kinase (PKA), potentiated the effects of 1alpha,25-dihydroxyvitamin D3(1,25-(OH)2D3)-induced 24-hydroxylation activity in HL-60 cells by increasing 1,25-(OH)2D3 receptor (VDR). The present study demonstrated that 12-O tetradecanoylphorbol-13-acetate (TPA), a potent phorbol ester, also potentiated the effect of 1,25-(OH)2D3 on HL-60 cells and that TPA and dbcAMP acted in a synergistic manner to enhance the effect of 1,25-(OH)2D3. It is interesting that TPA induced 24-hydroxylation activity far more efficiently than dbcAMP, in addition to their effects in increasing VDR. TPA increased basal levels of c-fos mRNA to the maximum by 1 h after the treatment, whereas dbcAMP failed to affect c fos gene expression. Together with the previous data indicating the presence of AP-1-like sequence in the promoter of 24-hydroxylase gene, it was suggested that TPA potentiated the effect of 1,25-(OH)2D3 through an activation of c-fos gene expression. This notion was further supported by the data showing that TPA and dbcAMP also acted in a synergistic manner to activate c-fos gene expression. Neither TPA nor dbcAMP affected c-jun early response gene in the HL-60 clone used in the present study. The present study suggested that the activation of early c fos response gene by TPA might be another mechanism to enhance the effect of 1,25 (OH)2D3, besides up-regulation of VDR. PMID- 10460018 TI - A novel germline mutation of multiple endocrine neoplasia type 1 (MEN1) gene in a Japanese MEN1 patient and her daughter. AB - Familial multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant inherited disorder characterized by tumors of the parathyroid, anterior pituitary and gastro-entero-pancreatic endocrine tissues. The MEN1 gene has recently been cloned and its germline mutations have been considered to play an important role in the tumorigenesis of MEN1. We analyzed a Japanese MEN1 patient and her daughter for germline mutations of the MEN1 gene. The proband (60 y.o.) had primary hyperparathyroidism (PHP) and gastrinoma, and her daughter (30 y.o.) had prolactinoma. Clinical examinations revealed no evidence of PHP in the daughter. We identified a novel heterozygous germline mutation (712 A del) at codon 201 in exon 3 of the MEN1 gene in the proband. Restriction digestion analysis revealed the same mutation pattern in her daughter. These findings suggest that this family has familial MEN1 including a rare case of MEN1 with a single lesion of the pituitary. Genetic examinations are useful as diagnostic tools for any rare or variant case of familial MEN1. PMID- 10460019 TI - Marked increase in plasma ACTH with tumor reduction after chemotherapy in ectopic ACTH syndrome. AB - We report on a case of rapid and marked hormone release as a result of rapid tumor reduction due to chemotherapy in a 36-year-old woman with ectopic ACTH syndrome due to small cell lung cancer. Treatment of the cancer with cisplatin and etoposide resulted in an 80% reduction in tumor size on computed tomographic scan within two weeks. Concurrently, plasma ACTH exhibited an unexpected and astonishing increase from 373 pg/ml before treatment to more than 1200 pg/ml. There were no biochemical characteristics observed in tumor lysis syndrome of solid tumors such as azotemia, increased LDH and hyperkalemia. The present case indicates that anticancer chemotherapy instituted in patients with ectopic ACTH syndrome could result in an acute increase of plasma ACTH and exacerbation of hypercortisolism, similar to tumor lysis syndrome, which is a potentially fatal complication following anti-cancer chemotherapy. PMID- 10460020 TI - Changes in serum levels of type I collagen-related proteins after surgically induced menopause and correlations with bone loss in the lumbar spine. AB - The purpose of this prospective study was to characterize the changes in serum levels of two proteins produced during the synthesis and degradation of type I collagen, i.e., the carboxyterminal propeptide of type I procollagen (PICP) and the pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP), respectively, after oophorectomy, and to assess the degree of correlation between changes in the serum values of these proteins and changes in bone mineral density (BMD) of the lumbar spine. Serum levels of PICP, ICTP and bone gla protein (BGP) were determined in 18 women before oophorectomy (baseline) and at 7 days, and 1, 2, 3, 6, 9 and 12 months post-oophorectomy (PO). The BMD of the lumbar spine was measured at baseline, and at 6 months and 12 months PO. ICTP had increased significantly at 7 days PO and peaked between 1 and 3 months PO. PICP and BGP had increased significantly at 2 months PO and remained at high levels thereafter. The percent changes in lumbar BMD from baseline values (% CFB) at 6 months and at 12 months PO were significantly correlated with % CFB in ICTP, but not with % CFB in PICP or BGP. Accordingly, bone resorption is a main determinant of bone mineral loss after oophorectomy and the change in recently-developed bone resorption markers, such as ICTP, is of clinical utility in predicting a degree of subsequent bone loss after surgical menopause. PMID- 10460021 TI - A patient with GH-producing pituitary adenoma presenting with a binasal superior quadrantanopsia. PMID- 10460022 TI - Low cholesterol in severe hypothyroidism? PMID- 10460023 TI - 'Pet Travel Scheme': the Government starts to publicise the requirements. PMID- 10460024 TI - Growing concern over TB in cattle. PMID- 10460026 TI - Efficacy of a single dose of oral antibiotic given within two hours of birth in preventing watery mouth disease and illthrift in colostrum-deficient lambs. AB - An antibiotic with a product licence limited to the treatment and control of infectious bacterial enteritis associated with Escherichia coli in piglets was tested for its ability to control watery mouth disease in neonatal lambs. Three groups of lambs were kept in conditions commonly encountered in intensive lambing systems, where high levels of environmental bacterial contamination may be expected. They were allocated at birth to: a control group (group 1) consisting of 18 colostrum-deprived lambs; group 2, consisting of 17 lambs given one feed of colostrum when they were two hours old; and group 3, consisting of 18 colostrum deprived lambs given spectinomycin orally when they were two hours old. Nine group 1 lambs became diseased and were killed for humane reasons. Blood biochemical changes included hyperglycaemia followed by hypoglycaemia, lactacidaemia, hypoproteinaemia and metabolic acidosis, and postmortem examination of the diseased lambs showed signs consistent with endotoxaemia and a clinical diagnosis of watery mouth disease. Coliforms were isolated from the blood of all group 1 lambs and from half the lambs in groups 2 and 3, but endotoxaemia and watery mouth disease occurred only in group 1 lambs. The results for groups 2 and 3 showed that neither colostrum nor antibiotic at the rates and frequency used prevented bacteraemia, although consecutive samples were positive only on three occasions. Group 3 lambs consistently grew more rapidly than the surviving group 1 lambs and as rapidly as group 2 lambs. There was no evidence that male lambs were more prone to watery mouth disease than female lambs. The results indicated that the antibiotic spectinomycin did not induce endotoxaemia during low-grade bacteraemia and that a single oral dose given within two hours of birth protected colostrum-deprived lambs delivered into a contaminated indoor environment against watery mouth disease. PMID- 10460027 TI - Seroprevalence of Neospora caninum and abortion in dairy cows in northern Spain. AB - The seroprevalence of Neospora caninum infection was estimated from a sample of 889 cattle from 43 dairy herds in three counties in the Asturias region of Spain. The true prevalence of infection was estimated to be 30.6 per cent (95 per cent confidence interval (CI) 27.6 to 33.6). Seropositivity was associated with abortion during the previous year (odds ratio (OR)=3.31, P<0.001) and was slightly higher among purchased cattle (37.6 per cent), than among cattle raised on the farm (29.1 per cent) (P=0.078). Seropositive cows were more likely than seronegative cows to have had a seropositive dam (OR=2.3, P=0.011), suggesting that congenital transmission contributed to about 56 per cent of the infections. Herds with a true seroprevalence above 10 per cent had more dogs on the farm, than herds with a lower prevalence (P=0.032). The ORS relating abortion to seropositivity in individual herds ranged from 0.7 to 19, indicating that some herds experienced few abortions caused by N. caninum, while others experienced more abortions due to the organism. Overall, 38.7 per cent of the abortions were estimated to have been attributable to N. caninum. PMID- 10460028 TI - Persistent vitelline vein in a foal. AB - A three-day-old foal died from intestinal strangulation due to a remnant of vitelline vein which extended between the umbilicus and the portal vein. The strangulating vein was identified on the basis of its morphological and histological structure. This finding, which is the first reported case of a persistent vitelline vein in a horse, is discussed in relation to the normal development and involution of the vitelline circulation. PMID- 10460029 TI - Possibility of sexual transmission of foot-and-mouth disease from African buffalo to cattle. PMID- 10460030 TI - Evidence of neoplastic nature and viral aetiology of so-called fowl glioma. PMID- 10460031 TI - Multiple cutaneous mast cell tumour in a calf. PMID- 10460032 TI - Outbreak of Listeria ivanovii abortion in sheep in India. PMID- 10460033 TI - Anaesthesia of cranes with alphaxolone-alphadolone. PMID- 10460034 TI - Borna disease in cats. PMID- 10460035 TI - Withdrawal of Immobilon LA. PMID- 10460036 TI - Post-weaning multisystemic wasting syndrome in Spain. PMID- 10460037 TI - Balance of the sexes. PMID- 10460038 TI - Balance of the sexes. PMID- 10460039 TI - The internal medicine clerkship: the view from the vantage point of one chair of medicine. PMID- 10460040 TI - Interferon alpha-2B and ribavirin in combination for patients with chronic hepatitis C who failed to respond to, or relapsed after, interferon alpha therapy: a randomized trial. AB - PURPOSE: To assess the efficacy of interferon alpha-2b and ribavirin in combination in the treatment of patients with chronic hepatitis C who had either failed to respond to therapy with interferon alpha (nonresponders), or who had relapsed after interferon therapy (relapsers). SUBJECTS AND METHODS: Four hundred patients with chronic hepatitis C (200 nonresponders and 200 relapsers) were randomly assigned in equal numbers to receive either subcutaneous administration of recombinant interferon alpha-2b (3 million units three times per week) and ribavirin (1,000 to 1,200 mg/daily orally) or interferon alpha-2b alone (6 million units three times per week). Both ribavirin and interferon alpha-2b were given for 24 weeks. The patients were then followed for an additional 24 weeks. RESULTS: At the end of the treatment period, normalization of serum alanine aminotransferase levels and absence of hepatitis C virus RNA were seen in 21% of nonresponders and in 39% of relapsers who were treated with interferon alpha-2b and ribavirin, compared with 5% of nonresponders (P = 0.001) and 9% of relapsers treated with interferon alpha-2b alone (P <0.001). At the end of follow-up, 14% of nonresponders and 30% of relapsers treated with the combination therapy had a sustained response, compared with 1% of nonresponders (P = 0.001) and 5% of relapsers treated with interferon alpha alone (P <0.001). CONCLUSIONS: A 24-week course of treatment with interferon alpha-2b and ribavirin offers a chance of sustained response, whereas retreatment with interferon alpha-2b alone does not give satisfactory results. The role of long-term therapy in inducing prolonged remission remains to be explored. PMID- 10460041 TI - A randomized trial of povidone-iodine compared with iodine tincture for venipuncture site disinfection: effects on rates of blood culture contamination. AB - BACKGROUND: Contamination of blood cultures creates problems in their interpretation and unneeded resource utilization. Because skin flora comprise the major group of contaminant species, more effective skin disinfection at the venipuncture site could reduce contamination. SUBJECTS AND METHODS: We performed a randomized trial in adult inpatients at a tertiary care teaching hospital. Antecubital venipuncture sites were randomly disinfected with povidone-iodine or iodine tincture, and blood cultures (two bottles, 10 mL of blood) were drawn by professional phlebotomists. Scoring of contaminant species was restricted to skin flora. Hospital resource utilization was compared among patients with contaminated blood cultures and those with sterile blood cultures. RESULTS: Of the 3,851 blood cultures collected during the study, 120 (3.1%) were contaminated with skin flora. The contamination rate for blood cultures collected after povidone-iodine was 3.8% (74 of 1,947), compared with a rate of 2.4% (46 of 1,904, P = 0.01) after iodine tincture. The difference in mean total hospital costs for patients with contaminated blood cultures and those with sterile blood cultures was $4,100 (95% confidence interval: $740 to $7,400, P = 0.02). CONCLUSIONS: Iodine tincture is superior to povidone-iodine for venipuncture site antisepsis before blood culture sampling. Because of the high costs associated with contaminated blood cultures, hospitals should consider switching from povidone-iodine to iodine tincture. Reduction of the contamination rate may improve the quality of patient care and reduce hospital costs. PMID- 10460042 TI - The use of a transscrotal testosterone delivery system in the treatment of patients with weight loss related to human immunodeficiency virus infection. AB - PURPOSE: Weight loss is a strong predictor of morbidity and mortality in human immunodeficiency virus (HIV)-infected patients. Men with acquired immunodeficiency syndrome (AIDS) lose body cell mass. Hypogonadism is also common. This study tested the efficacy of a testosterone transscrotal patch (6 mg/day) in improving body cell mass and treating hypogonadism in these patients. SUBJECTS AND METHODS: This multicenter, randomized, double-blinded, placebo controlled trial was conducted from August 1995 to October 1996 in 133 men, 18 years of age and older, who had AIDS, 5% to 20% weight loss, and either a low morning serum total testosterone level (<400 ng/dL) or a low free testosterone level (<16 pg/mL). Outcomes included weight, body cell mass as measured using bioelectrical impedance analysis, quality of life, and morning measurements of serum testosterone and dihydrotestosterone levels, lymphocyte subsets, and HIV quantification. RESULTS: There were no significant differences in baseline weight, CD4 cell counts, or HIV serum viral quantification between treatment arms. Morning total and free testosterone levels increased in those treated with testosterone, but not with placebo. Following 12 weeks of treatment there were no differences (testosterone-placebo) in mean weight change (-0.3 kg [95% confidence interval (CI): -1.4 to 0.8]) or body cell mass (-0.2 kg [95% CI: -1.0 to 0.6]) in the two groups. There were also no changes in quality of life in either group. CONCLUSION: Hypogonadal men with AIDS and weight loss can achieve adequate morning serum sex hormone levels using a transscrotal testosterone patch. However, this system of replacement does not improve weight, body cell mass, or quality of life. PMID- 10460043 TI - Symptom status and the desire for Helicobacter pylori confirmatory testing after eradication therapy in patients with peptic ulcer disease. AB - BACKGROUND: There is consensus that individuals with Helicobacter pylori associated peptic ulcer disease should receive a test to confirm H. pylori cure if symptoms recur after eradication therapy. It remains controversial whether individuals who are asymptomatic after therapy should undergo a confirmatory test to establish cure. Patients' desire to know whether their infection has been cured and symptom status after treatment are two important determinants of whether confirmatory H. pylori testing should be undertaken routinely. METHODS: We identified 87 patients with H. pylori-associated peptic ulcer disease scheduled to undergo urea breath testing 4 weeks after H. pylori eradication therapy. At the time of testing, willingness-to-pay methodology was used to estimate patients' desire for confirmatory testing in the absence of symptoms. At a follow-up visit after eradication therapy (mean follow-up, 297 days; range, 143 to 494 days), patients were surveyed to assess gastrointestinal symptom status. RESULTS: Of the 87 patients, 78 (90%) responded that they would prefer to undergo confirmatory testing if asymptomatic, as opposed to delaying testing until symptoms recurred. Patients' median willingness to pay for confirmatory testing in the absence of symptoms was greater than $50. On follow-up, 38% of patients in whom H. pylori was eradicated reported that their symptoms were completely resolved. There was no significant difference in the percentage of patients who reported complete symptom resolution by H. pylori status (H. pylori eradicated 38%, H. pylori infected 28%, P = 0.42, 95% confidence interval, -14% to 34%). CONCLUSIONS: Patients' desire for confirmation of cure, coupled with a frequent need for confirmatory testing as a result of recurrent symptoms after therapy, may justify routine confirmatory testing after H. pylori treatment. PMID- 10460044 TI - The effect of hospital experience on mortality among patients hospitalized with acquired immunodeficiency syndrome in California. AB - BACKGROUND: Several studies in the 1980s suggested that mortality rates for patients hospitalized with the acquired immunodeficiency syndrome (AIDS) were lower in hospitals that cared for greater numbers of AIDS patients. We sought to determine whether this observation persisted in the mid-1990s in California. SUBJECTS AND METHODS: We performed an analysis of hospital discharge data for 7,901 adults discharged with human immunodeficiency virus (HIV) or AIDS-related diagnoses from all acute care hospitals (n = 333) in California during 1994. The main outcome measure was in-hospital mortality, adjusted for severity of illness, comorbidity, prior hospitalizations, and other patient and hospital characteristics. RESULTS: Among 7,901 persons hospitalized with AIDS, the unadjusted inpatient mortality was 9.0%. The adjusted mortality rate varied significantly (P <0.0001) from 12.4% among institutions with the lowest quartile of AIDS experience to 10.3%, 6.3%, and 7.6% by quartile of greater AIDS experience. Increasing severity of illness, comorbidity, and previous hospitalizations were also significant predictors of increased mortality. Sex, race, and insurance status were not associated with hospital mortality. CONCLUSIONS: Hospital mortality for AIDS patients was greater among less experienced hospitals. The difference in mortality rate was equivalent to more than four additional deaths per 100 patients with AIDS admitted to the least experienced hospitals. This finding was not explained by severity of illness, comorbidity, or other case-mix variables. PMID- 10460045 TI - Clinical features and costs of care for hospitalized adults with primary Epstein Barr virus infection. AB - PURPOSE: To study the clinical and laboratory features of primary infection with the Epstein-Barr virus in adults who required hospitalization and to assess the difficulty in its diagnosis, the use of diagnostic procedures, and the associated costs of care. PATIENTS AND METHODS: We retrospectively identified all adult patients who were diagnosed with primary Epstein-Barr virus infection in our region between 1988 and 1997 using strict serologic criteria. The added costs of unnecessary diagnostic tests and treatment were estimated. RESULTS: The analysis included 47 patients (60% men) with a mean (+/-SD) age of 30 +/- 14 years. The prime cause of admission was fever (83%). Compared with patients 35 years of age and older, those younger than 35 years were more likely to have pharyngitis (45% vs 10%) and lymphadenopathy (66% vs 17%). Younger patients also had a greater mean atypical lymphocyte count (17% +/- 14% vs 8% +/- 6%) and more abnormal hepatic enzyme levels. Inpatient work-ups resulted in 309 days of hospitalization, many diagnostic tests, and unnecessary empiric treatments (total 203 days of antibiotic therapy). Overall, unnecessary diagnostic procedures and medical treatments contributed an average of approximately $12,000 in health-care costs per patient. CONCLUSIONS: Testing for primary Epstein-Barr virus infection should be a routine step in the investigation of fever in adults of all ages. A higher index of suspicion might prevent unnecessary, sometimes hazardous inpatient work-ups from being performed, thereby reducing health-care expenses. PMID- 10460046 TI - Encounters with pharmaceutical sales representatives among practicing internists. AB - BACKGROUND: Although pharmaceutical sales representatives provide physicians with information on new products, these encounters have rarely been studied in practice settings. We examined these interactions among practicing internists and assessed whether prior residency policies limiting pharmaceutical sales representative access affected the subsequent behavior of practitioners. METHODS: We conducted a mail survey of the internal medicine staffs of a medical school hospital and two affiliated community hospitals. A second request was sent to nonresponders. After the second mailing, a random sample of nonresponders was compared with a similar sample of respondents. Multivariate odds ratios (OR) and 95% confidence intervals (CI) were estimated with logistic regression. RESULTS: Of the 346 (40%) internists who responded, 22% were women and 60% were trained in university hospitals. There were no differences in gender, subspecialization, or type of training when survey responders and nonresponders were compared. Two hundred eighty-seven (83%) physicians had met with pharmaceutical sales representatives within the previous year, of whom 248 (86%) had received drug samples. Having had a policy that limited access to pharmaceutical sales representatives during residency did not affect the subsequent likelihood of seeing these representatives (P = 0.20) or accepting samples in practice (P = 0.99). Those describing themselves as busy practitioners were significantly less likely to abstain from meeting pharmaceutical sales representatives (OR = 0.2, 95% CI: 0.1 to 0.6, P <0.001). Those with very frequent contacts (>10 times/month) were virtually all busy practitioners. CONCLUSIONS: Encounters between physicians and pharmaceutical sales representatives are common in internal medicine practice, especially in busy offices. Policies designed to limit pharmaceutical sales representative access during residency do not appear to affect the subsequent likelihood of meeting with pharmaceutical sales representatives or accepting samples. PMID- 10460047 TI - The sequence of withdrawing life-sustaining treatment from patients. AB - PURPOSE: To describe the observed sequence of withdrawal of eight different forms of life-sustaining treatment and to determine whether aspects of those treatments determine the order of withdrawal. SUBJECTS AND METHODS: We observed 211 consecutive patients dying in four midwestern US hospitals from whom at least one of eight specific life-sustaining treatments was or could have been withdrawn. We used a parametric statistical technique to explain the order of withdrawal based on selected characteristics of the forms of life support, including cost, scarcity, and discomfort. RESULTS: The eight forms of life support were withdrawn in a distinct sequence. From earliest to latest, the order was blood products, hemodialysis, vasopressors, mechanical ventilation, total parenteral nutrition, antibiotics, intravenous fluids, and tube feedings (P <0.0001). The sequence was almost identical to that observed in a previous study based on hypothetical scenarios. Forms of life support that were perceived as more artificial, scarce, or expensive were withdrawn earlier. CONCLUSION: The preference for withdrawing some forms of life-sustaining treatments more than others is associated with intrinsic characteristics of these treatments. Once the decision has been made to forgo life-sustaining treatment, the process remains complex and appears to target many different goals simultaneously. PMID- 10460048 TI - Leukocyte adhesion molecules in transplantation. PMID- 10460049 TI - Penicillin skin testing: a way to optimize antibiotic utilization. PMID- 10460050 TI - Intravascular lymphoma associated with endocrine dysfunction: a report of four cases and a review of the literature. PMID- 10460051 TI - Acute monoarthritis from infectious mononucleosis. PMID- 10460052 TI - Treatment of hepatitis C: a decision that needs careful consideration. PMID- 10460053 TI - Interactions between physicians and drug company representatives. PMID- 10460054 TI - A pilot study of the use of smokeless tobacco. PMID- 10460055 TI - A pilot study of the use of smokeless tobacco. PMID- 10460057 TI - Castleman's disease with Kaposi's sarcoma and glomerulonephritis. PMID- 10460056 TI - Intestinal involvement in Hodgkin's disease causing perforation. PMID- 10460058 TI - A mesenteric mass in a chronic LSD user. PMID- 10460059 TI - Magnesium in mineral bottled waters and cerebrovascular stroke. PMID- 10460060 TI - Diagnostic yield of pericardial drainage depends on underlying diseases in the population. PMID- 10460061 TI - Beta blocker therapy in cholinergic urticaria. PMID- 10460062 TI - Expulsion of an artificial eye in a patient with thyroid-associated ophthalmopathy and surgical anophthalmos. PMID- 10460063 TI - New frontiers of cardioprotection. PMID- 10460064 TI - Propofol decreases the clearance of midazolam by inhibiting CYP3A4: an in vivo and in vitro study. AB - OBJECTIVE: To examine the effect of propofol on the pharmacokinetics of midazolam in vivo and to elucidate the mechanism of the pharmacokinetic changes of midazolam by propofol with the use of human liver microsomes and recombinant CYP3A4. METHODS: In an in vivo, double-blind randomized study, 24 patients received 0.2 mg/kg midazolam and either 2 mg/kg propofol (propofol group) or placebo (placebo group) for induction of anesthesia. In the propofol group, continuous infusion of propofol at 9 mg/kg/h was started immediately after the bolus infusion of propofol and was maintained for an hour. In the placebo group the same dose of soybean emulsion as a placebo was given and infused intravenously for an hour instead of propofol. In an in vitro study the effect of propofol on the metabolism of midazolam was studied with human liver microsomes and recombinant CYP3A4. RESULTS: In the propofol group the mean clearance of midazolam was decreased by 37% (P = .005) and the mean elimination half-life was prolonged by 61% (P = .04) compared with the placebo group. The mean plasma concentrations of 1'-hydroxymidazolam were lower in the propofol group than in the placebo group at 5, 10, 15, 20, and 30 minutes after midazolam was administered (P < .05). The mean (+/-SD) Michaelis-Menten constant for midazolam 1'-hydroxylation by human liver microsomes was 5.6 +/- 3.3 micromol/L. The formation of 1'-hydroxymidazolam was competitively inhibited by propofol, and the mean inhibition constant was 56.7 +/- 16.6 micromol/L. The mean Michaelis-Menten constant and mean inhibition constant values for midazolam 1'-hydroxylation by recombinant CYP3A4 were 4.0 micromol/L and 61.0 micromol/L, respectively, consistent with the mean values obtained from human liver microsomes. CONCLUSION: Propofol decreases the clearance of midazolam, and the possible mechanism is the competitive inhibition of hepatic CYP3A4. PMID- 10460065 TI - Grapefruit juice increases serum concentrations of atorvastatin and has no effect on pravastatin. AB - BACKGROUND: Grapefruit juice greatly increases the bioavailability of lovastatin and simvastatin. We studied the effect of grapefruit juice on the pharmacokinetics of atorvastatin and pravastatin. METHODS: Two randomized, two phase crossover studies were performed--study I with atorvastatin in 12 healthy volunteers and study II with pravastatin in 11 healthy volunteers. In both studies, volunteers took 200 mL double-strength grapefruit juice or water three times a day for 2 days. On day 3, each subject ingested a single 40 mg dose of atorvastatin (study I) or pravastatin (study II) with either 200 mL grapefruit juice or water, and an additional 200 mL was ingested 1/2 hour and 1 1/2 hours later. In addition, subjects took 200 mL grapefruit juice or water three times a day on days 4 and 5 in study I. In study I, serum concentrations of atorvastatin acid, atorvastatin lactone, 2-hydroxyatorvastatin acid, 2-hydroxyatorvastatin lactone, and active and total 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors were measured up to 72 hours. In study II, pravastatin, pravastatin lactone, and active and total HMG-CoA reductase inhibitors were measured up to 24 hours. RESULTS: Grapefruit juice increased the area under the serum concentration-time curve of atorvastatin acid from time zero to 72 hours [AUC(0-72)] 2.5-fold (P < .01), whereas the peak serum concentration (Cmax) was not significantly changed. The time of the peak concentration (tmax) and the elimination half-life (t1/2) of atorvastatin acid were increased (P < .01). The AUC(0-72) of atorvastatin lactone was increased 3.3-fold (P < .01) and the Cmax 2.6-fold (P < .01) by grapefruit juice, and the tmax and t1/2 were also increased (P < .05). Grapefruit juice decreased the Cmax (P < .001) and AUC(0-72) (P < .001) of 2-hydroxyatorvastatin acid and increased its tmax and t1/2 (P < .01). Grapefruit juice also decreased the Cmax (P < .001) and AUC(O-72) (P < .05) of 2 hydroxyatorvastatin lactone. The AUC(0-72) values of active and total HMG-CoA reductase inhibitors were increased 1.3-fold (P < .05) and 1.5-fold (P < .01), respectively, by grapefruit juice. In study II, the only significant change observed in the pharmacokinetics of pravastatin was prolongation of the tmax of active HMG-CoA reductase inhibitors by grapefruit juice (P < .05). CONCLUSIONS: Grapefruit juice significantly increased serum concentrations of atorvastatin acid, atorvastatin lactone, and active and total HMG-CoA reductase inhibitors, probably by decreasing CYP3A4-mediated first-pass metabolism of atorvastatin in the small intestine. On the other hand, grapefruit juice had no effect on the pharmacokinetics of pravastatin. Concomitant use of atorvastatin and at least large amounts of grapefruit juice should be avoided, or the dose of atorvastatin should be reduced accordingly. PMID- 10460066 TI - Disposition kinetics and effects of menthol. AB - BACKGROUND: Menthol is widely used in a variety of commercial products and foods, but its clinical pharmacology is not well studied. To determine the disposition kinetics and to examine subjective and cardiovascular effects of menthol, we conducted a crossover placebo-controlled study that compared pure menthol versus placebo, along with an uncontrolled exposure to menthol in food or beverage. A novel assay for the measurement of menthol in biological fluids was also developed. METHODS: Twelve subjects were studied; each received a 100 mg l menthol capsule, a placebo capsule, and 10 mg menthol in mint candy or mint tea on three different occasions. Plasma and urine levels of menthol and conjugated menthol (glucuronide), cardiovascular measurements, and subjective effects were measured at frequent intervals. RESULTS: Menthol was rapidly metabolized, and only menthol glucuronide could be measured in plasma or urine. The plasma half life of menthol glucuronide averaged 56.2 minutes (95% confidence interval [CI], 51.0 to 61.5) and 42.6 minutes (95% CI, 32.5 to 52.7) in menthol capsule and mint candy/mint tea conditions, respectively (P < .05). The plasma area under the plasma concentration-time curve ratios for menthol capsule to mint candy/mint tea treatment averaged 9.2 (95% CI, 8.2 to 10.1). Urinary recovery of menthol as the glucuronide averaged 45.6 and 56.6% for menthol capsule and mint candy/tea, respectively (difference not significant). After menthol capsule dosing, the decrease in heart rate was less than the decrease after placebo administration (P < .05). Menthol reduced subjective vigor value at 30 minutes. CONCLUSIONS: We conclude that pure menthol and menthol in food or beverages have a similar systemic bioavailability and that menthol has a small cardioaccelerating effect. PMID- 10460067 TI - Pharmacokinetics and tolerance of flunitrazepam in neonates and in infants. AB - OBJECTIVE: To study the pharmacokinetics of flunitrazepam (used for sedation in neonates and infants), to determine the influence of both gestational and postnatal age on the pharmacokinetic parameters, and to analyze the relationship between the hemodynamic parameters and flunitrazepam plasma concentration. METHODS: Flunitrazepam was infused for 20 minutes as a single dose (0.2 mg x kg( 1)) and as multiple doses (0.1 mg x kg(-1)). Six to eight 1-mL blood samples were collected per patient. Flunitrazepam plasma concentration was measured by gas chromatography-mass spectrometry. RESULTS: Thirty-one patients (25 neonates and six infants) were included in the study. Only three of them received multiple doses. After the single dose (n = 28), half-life was 22.6 +/- 7.3 hours, clearance was 0.15 +/- 0.14 L x kg x h(-1), and volume of distribution was 4.6 +/ 4.1 L x kg(-1) (mean +/- SD). Plasma clearance and volume of distribution significantly increased with postnatal age (P < .05), but no pharmacokinetic parameter varied significantly with gestational age. Diastolic blood pressure significantly decreased with increasing flunitrazepam plasma concentrations (P < .05). CONCLUSION: Postnatal age but not gestational age influenced flunitrazepam pharmacokinetic parameters in neonates and infants. Diastolic blood pressure was inversely correlated to flunitrazepam plasma concentration. PMID- 10460068 TI - Pharmacodynamic modeling of the acid inhibitory effect of ranitidine in patients in an intensive care unit during prolonged dosing: characterization of tolerance. AB - OBJECTIVE: To characterize the relationship between the pharmacokinetics and the acid inhibitory effect of ranitidine during prolonged dosing on the basis of a physiologic indirect-response model. METHODS: Multiple doses of ranitidine were administered to 18 patients in an intensive care unit in an open randomized trial. All patients received an initial intravenous dose of 50 mg ranitidine; after 12 hours repeated injections (50 mg every 6 hours) or a primed continuous infusion (50 mg plus 0.125 mg/kg/h) was administered. Intragastric pH was monitored continuously for at least 42 hours. RESULTS: After the initial injection a time lag was observed between the increase of plasma concentration and the increase of pH. With the indirect-response model the rate of onset of effect (kout) could be estimated adequately by relating the inhibitory effect on acid secretion to the concentration according to a sigmoid Emax model. For administration of a single dose, estimated pharmacodynamic parameters were 4.5 +/ 0.9 h(-1) for kout, 1.4 +/- 0.1 for baseline pH, 0.051 +/- 0.012 mg/L for 50% inhibition constant, and 7.0 +/- 1.5 for Hill factor (mean +/- SEM; n = 18). Tolerance developed during subsequent dosing that could be described as a linear increase (beta) of 50% inhibition constant with time (beta = 0.0030 and 0.0045 mg/L/h for repeated and continuous administration, respectively). CONCLUSIONS: The developed physiologic indirect-response model may be used to quantify the pharmacokinetic-pharmacodynamic relationship of ranitidine during single and multiple dosing. During prolonged intravenous dosing, tolerance developed within 42 hours and could be characterized on the basis of the developed indirect response model. PMID- 10460069 TI - Clomipramine dose-effect study in patients with depression: clinical end points and pharmacokinetics. Danish University Antidepressant Group (DUAG). AB - OBJECTIVE: To examine the problems of establishing dose-effect and concentration effect relationships of antidepressant therapy with clomipramine. METHODS: This randomized double-blind study compared five fixed doses of clomipramine hydrochloride: 25, 50, 75, 125, and 200 mg/day in hospitalized or day patients at nine clinical centers in Denmark. A 1-week washout period was followed by 6 weeks of active treatment and weekly depression ratings. In total, 151 patients (100 women and 51 men) with major depression scoring > or =18 on the Hamilton Depression Scale (HDS) or > or =9 on the Hamilton Depression subscale (HDSS) before and after the washout period were randomized. The treatment groups (n = 29 to 32) were well balanced with respect to sex, age, and depression rating. Serum concentrations of clomipramine plus metabolites were measured at weekly intervals. A sparteine test was performed before and during drug treatment. RESULTS: There was pronounced interpatient variability in response and kinetics at each dose. Drop-outs attributable to adverse events increased with rising doses, whereas drop-outs caused by worsening or lack of effect or nonresponse declined with increasing dose. Completer analyses showed a moderate and statistically significant relationship between depression rating and dose at all ratings after 1 to 6 weeks of treatment (trend analysis). HDS items representing core symptoms of depression showed a particularly consistent dose-effect relationship. Early sustained response occurred more frequently with the two highest doses. Serum levels of clomipramine and desmethylclomipramine showed weak correlation with depression ratings (Rs = -0.18 to -0.27; P < .05 to P < .01). A few blood pressure measurements and a few typical side-effect ratings showed a statistically significant dose-effect and concentration-effect relationship. Serum concentration of clomipramine and desmethylclomipramine showed a pronounced disproportionate increase with increasing dose. Clomipramine inhibited in a dose dependent fashion CYP2D6 (sparteine oxidation). CONCLUSION: The dose-effect curves, indicating the probability of a certain outcome at a given dose, were flat and overlapping suggesting a narrow therapeutic range. This pattern is similar to that observed with newer antidepressants. PMID- 10460070 TI - Fenofibrate decreases plasma fibrinogen, improves lipid profile, and reduces uricemia. AB - Plasma fibrinogen has been found to be a major cardiovascular disease risk factor. This 2-year trial was designed to assess the effect of fenofibrate on fibrinogen and, as secondary end points, on lipid profile and uric acid in patients with dyslipidemia. Eighty subjects (40 women and 40 men) were admitted to either a control or an active group. Sixty-seven (84%) had sole hypercholesterolemia, 13 (16%) subjects had mixed dyslipidemia. The effect attributable to fenofibrate was a decrease of 15% in fibrinogen, 26% in the ratio low-density lipoprotein cholesterol to high-density lipoprotein cholesterol (-20% low-density lipoprotein cholesterol, +10% high-density lipoprotein cholesterol), 34% in triglycerides (median), and 13% in uric acid (P < .0001 for all). Fenofibrate simultaneously affected hemostasis (by lowering fibrinogenemia) and lipid profile. Because fenofibrate has few adverse effects, it could be a fair option for patients who need polytherapy and do not tolerate resins or niacin. Its clinical efficacy should be tested in long-term studies to assess its real capacity to prevent cardiovascular events. PMID- 10460071 TI - Implication of evidence-based medicine in prescription guidelines taught to French medical students: current status in the cardiovascular field. AB - OBJECTIVES: To study how satisfactory the contents of introductory courses in cardiovascular therapeutics, given to medical students in France, are with respect to the concepts of evidence-based medicine. METHODS: Medical school lecturers were asked to provide written course material used as part of medical school courses. Best-available evidence was classified as existent (including two therapeutic subclasses: indicated and contraindicated), and nonexistent. Four scores (from 0 to 10) were given, according to conformity with best-available evidence, and citation of randomized clinical trials (RCT), meta-analyses and therapeutic objectives. RESULTS: Thirty-four written documents were obtained from 43 faculties. Although the score (mean +/- SEM) of conformity with best-available evidence was 5.43 +/- 0.28 for the existent best-available evidence class, the corresponding scores for the citation of RCT, meta-analyses, and therapeutic objectives were, respectively, 1 +/- 0.2, 0.16 +/- 0.07, and 2.7 +/- 0.3. The four scores were highest when the best-available evidence belonged to the indicated class, intermediate when best-available evidence was nonexistent, and lowest for the contraindicated class (P < .05). These scores were significantly higher when the printed material was intended for specialists. CONCLUSION: Despite some limitations, the extent of agreement with the best-available evidence is only moderate. Pathophysiologic reasoning is largely preferred to justify the choice of therapeutics recommended to medical students. PMID- 10460072 TI - Low frequency of defective alleles of cytochrome P450 enzymes 2C19 and 2D6 in the Turkish population. AB - BACKGROUND AND OBJECTIVES: The genetically polymorphic cytochrome P450 enzymes 2Cl9 (CYP2Cl9) and 2D6 (CYP2D6) contribute to the metabolism of about 30% of all drugs. For analysis of the ethnic-related differences in drug disposition and as a preparation for routine genotyping, we examined CYP2C19 and CYP2D6 mutations in a large Turkish population. METHODS: CYP2C19 and CYP2D6 alleles were determined with use of genomic deoxyribonucleic acid from 404 unrelated Turkish individuals. CYP2C19 alleles *1 to *5 and CYP2D6 alleles *1 to *12, and *14, *15, and *17 were measured by polymerase chain reaction-restriction fragment length polymorphism assays. RESULTS: From 404 subjects genotyped for CYP2C19, allele frequencies of CYP2C19*1 (wt), CYP2C19*2 (ml), and CYP2C19*3 (m2) were 0.88, 0.12, and 0.004, respectively; mutations m3 and m4 were not found. Four individuals (1.0%) were predicted to be poor metabolizers (CYP2C19*2/*2), a significantly lower frequency compared to Middle European populations. Among 404 subjects genotyped for CYP2D6, most frequent alleles were CYP2D6*1 (allele frequency 0.37), *2 (0.35), *4 (0.11), *10 (0.06), duplications *1x2, *2x2, or *4x2 (0.06), *5 (0.01), and *17(0.01). Overall, six subjects (1.49%) were predicted to be CYP2D6 poor metabolizers, and 35 subjects (8.66%) were predicted to be ultrarapid metabolizers as a result of CYP2D6 gene duplications. CONCLUSION: Obviously, within Europe there is a north-south gradient, with decreasing frequency of poor metabolizers of CYP2C19 and CYP2D6 to the south and a corresponding increase of ultrarapid metabolizers of CYP2D6. As in other white groups, only CYP2C19*2 plays a relevant role for the CYP2C19 poor metabolizer phenotype. The mutational spectrum of CYP2D6 indicated partial ethnic relationships to Asian and African populations. PMID- 10460073 TI - Polymorphism of dextromethorphan oxidation in South Indian subjects. AB - One hundred fifty-six unrelated healthy South Indian subjects were phenotyped according to their ability to metabolize dextromethorphan to its O-demethylated metabolite dextrorphan. Each volunteer was administered 25 mg oral dextromethorphan hydrobromide (19.3 mg dextromethorphan). Urine was collected during an 8-hour period after drug administration and was analyzed for dextromethorphan and dextrorphan by HPLC with fluorescence detection. This analysis was performed with and without previous deconjugation. The log10 (metabolic ratio), calculated as the ratio of dextromethorphan to dextrorphan, was bimodally distributed, and it was inferred that the frequency of occurrence of poor metabolizers of dextromethorphan in South Indian subjects is 3.2%. Phenotype assignment remained the same with both methods of analysis. Furthermore, a fairly good correlation (Spearman rank order correlation coefficient [r(s)] = 0.61; P < .0001) was observed between the log-transformed metabolic ratio derived from both methods. PMID- 10460074 TI - Dosing recommendations in liver disease. PMID- 10460075 TI - Variations in sedating uncooperative, stable children for post-traumatic head CT. AB - OBJECTIVE: To characterize variations among pediatric emergency physicians and their hospital facilities regarding sedation of the uncooperative, stable child for head CT following closed head injury. DESIGN: Mail survey with two follow-up mailings. PARTICIPANTS: Surveys were sent to all members of the Emergency Medicine Section of the American Academy of Pediatrics (AAP). RESULTS: Of 596 surveys sent, 431 (72%) were returned, with 304 (51%) usable responses. Respondents annually sedate over 17,500 children for post-traumatic head CT. Formal training to sedate children for head CT was noted by 73%. Published guidelines for sedation are followed by 74%; 10% were unaware of the existence of published guidelines for sedation. Twenty-six percent of the respondents were very or somewhat dissatisfied with their sedation-related practices. In response to three clinical scenarios involving sedation of 8-month-old, 3-year-old, and 6 year-old children for head CT, midazolam was the most commonly chosen drug. Over 20 different sedation strategies were selected for each scenario. CONCLUSIONS: Sedation practices for post-traumatic pediatric head CT vary widely, among both physicians and individual practitioners. Institutional and individual sedation relation policies vary widely as well. Variation and dissatisfaction with sedation practices may reflect uncertainty regarding optimal sedation strategies. Further cost-effectiveness research is necessary. PMID- 10460076 TI - Discordant radiograph interpretation between emergency physicians and radiologists in a pediatric emergency department. AB - OBJECTIVES: To describe the types of discrepancies in radiograph interpretation between emergency physicians and radiologists in a pediatric emergency department, and to determine the impact of discrepant interpretations on patient care. METHODS: Prospective cohort study of discordant radiographs from the period beginning March 1, 1995 and ending March 31, 1996. During this period, 2083 radiographs were coded by the radiologist as concordant or discordant. Three hundred forty-nine were coded as discordant, and 324 were eligible for the study. Charts were reviewed for relevant physical examination findings and emergency department management. Discrepancies that affected patient care were deemed clinically significant. RESULTS: Twenty-three (1.1%) of 2083 radiographs were interpreted differently by the emergency physician and the radiologist in a way that might have changed patient management. This represents 7% (23/324) of the radiographs originally coded by a radiologist as discrepant. The most common discrepancy was a patient with a normal chest examination and a radiograph interpreted as having an infiltrate by the emergency physician, but subsequently read as having no infiltrate by a radiologist (12/324). These patients may have received antibiotics unnecessarily. Two discrepant interpretations had the potential to have serious consequences to the patient if not identified. One patient with cardiomegaly and another patient with free air on abdominal radiograph were not noted by the emergency physician. CONCLUSIONS: Emergency physicians would benefit from more rigorous interpretation of chest x-rays to avoid unnecessary treatment with antibiotics. Emergency physicians do a good job interpreting plain radiographs, but occasionally miss significant findings that could lead to adverse outcomes. The presence of radiologists to immediately read radiographs 24 hours a day could prevent missed findings, but, given the small number of significant misinterpretations, is unlikely to be cost effective. PMID- 10460077 TI - Can portable bedside fluoroscopy replace standard, postreduction radiographs in the management of pediatric fractures? AB - OBJECTIVE: To determine the accuracy of portable bedside fluoroscopy in documenting postreduction fracture alignment in the pediatric emergency department (ED). DESIGN/SETTING: Prospective trial in an urban pediatric ED. PARTICIPANTS: Convenience sample of 80 pediatric patients requiring ED reduction of isolated long bone fractures. METHODS: Patients who underwent closed fracture reduction using portable fluoroscopic guidance (FluoroScan) in the ED were enrolled in the study. Postreduction images were obtained using both bedside fluoroscopy and conventional radiographs. A pediatric orthopedic subspecialist, blinded to clinical outcome, reviewed the fluoroscopic and radiographic images for adequacy of alignment and rated the utility of conventional radiography for fracture management. RESULTS: The patients were 2.5 to 16 years of age (mean 8.3). Distal radial and radioulnar fractures comprised 96% (76/80) of cases. Sixty-three percent of the fractures were displaced, and the mean angulation of the primary fracture site was 24 degrees . Fluoroscopy was found to be 100% sensitive (75/75 cases) and 100% specific (5/5 cases) in predicting postreduction fracture position when compared to conventional radiographs. Intra-rater observer agreement on the necessity of conventional postreduction radiographs was 0.92 (95% CI 0.82-1.00) using the kappa coefficient. In no case did postreduction radiographs alter acute fracture management. CONCLUSIONS: Bedside fluoroscopy with printed fluoroscopic images are highly reliable in evaluating fracture reduction and can replace conventional radiography in documenting adequate distal forearm fracture reduction when there is no intraarticular involvement. PMID- 10460078 TI - Otitis media in infants aged 0-8 weeks: frequency of associated serious bacterial disease. AB - OBJECTIVE: To determine the frequency of serious bacterial infection in well appearing infants aged 0-8 weeks with isolated otitis media (OM). METHODS: Infants with confirmed OM underwent tympanocentesis with middle ear fluid (MEF) culture and complete sepsis evaluation. Enrolled infants were admitted to the hospital for parenteral antibiotics until blood, urine, and CSF cultures were negative for 48 hours. RESULTS: Forty non-toxic appearing infants were enrolled between January 1994 and April 1995, of whom 15 (38%) had a documented rectal temperature > or =38 degrees C. Bacterial pathogens were isolated from MEF cultures in 25 (62.5%) infants. All afebrile infants had negative blood, urine, and cerebrospinal fluid cultures (upper limit (UL) 95% CI 0.11). Only two febrile infants had positive cultures from sites other than the MEF (UL 95% CI 0.36). CONCLUSION: In our study population, previously healthy, non-toxic appearing afebrile infants aged 2-8 weeks and having isolated OM infrequently have an associated serious bacterial infection, suggesting that outpatient treatment with oral antibiotics and close follow-up may be an option. Further studies with large numbers of infants are necessary to confirm this conclusion. PMID- 10460079 TI - Providing immunizations in a pediatric emergency department: underimmunization rates and parental acceptance. AB - OBJECTIVES: To assess the vaccination status and vaccinate eligible children with age-appropriate antigens. DESIGN: Intervention. SETTING: Pediatric emergency department in an urban, public hospital. PATIENTS: Convenience sample of children, aged birth through 72 months. INTERVENTIONS: Immunization of eligible children. MAIN OUTCOME MEASURES: 1) Immunization coverage rates in the sample population, 2) Acceptance rates of immunization. RESULTS: A total of 9321 children were enrolled over a 2-year period. Fifty-nine percent were documented to be underimmunized. Overall, 2514 children received a total of 6482 immunizations. Parents who carried portable immunization cards documenting that their child was underimmunized were almost five times more likely to accept immunization for their child than parents who lacked documentation (71% vs 15%, P < 0.0001). The estimated cost of providing immunizations in the emergency department was $47.15 per child immunized, or $18.56 per immunization given. CONCLUSIONS: The majority of children with documentation of immunization status were underimmunized. When documentation of underimmunization was available, parents were significantly more likely to accept vaccination. These data suggest that vaccinating children in nontraditional settings is feasible and support the creation of an accessible vaccine registry. PMID- 10460080 TI - Fatal laryngeal injury in an achondroplastic dwarf secondary to airbag deployment. AB - We report the case of an unrestrained driver with achondroplastic dwarfism who suffered a fatal anterior neck injury when her airbag deployed as she rear-ended another vehicle at 30-40 mph. Her short stature and short limbs required her to sit within a handbreadth of the steering wheel, which probably allowed the airbag or airbag cover to strike her neck as it opened. This is the first reported case of fatal injury to a driver with achondroplastic dwarfism. PMID- 10460081 TI - Acute dilated cardiomyopathy and central nervous system toxicity following propranolol intoxication. AB - OBJECTIVE: We report a case of a 16-year-old boy who developed central nervous system (CNS) depression and acute dilated cardiomyopathy following ingestion of 3200 mg of propranolol in a suicide attempt. Early echocardiographic findings were the only sign of cardiac toxicity. DESIGN: A case report. SETTING: Pediatric intensive care unit of a teaching hospital. RESULTS: This child developed significant acute dilated cardiomyopathy and severe CNS depression 2 hours after ingesting 3200 mg of propranolol. The child was treated with gastric lavage, activated charcoal, and mechanical ventilation. Following the echocardiographic findings, treatment with isoprenaline hydrochloride and glucagon were given intravenously. Echocardiographic examination 12 hours following treatment showed normal left ventricular size and function. No change in pulse rate or blood pressure was reported on admission and during his hospitalization. DISCUSSION: In the early stages of propranolol and other lipophilic beta-blocker intoxication, severe CNS depression can develop in the absence of clinical signs of cardiac toxicity. Early echocardiographic evaluation is important and may prevent delay in diagnosis and treatment of cardiac toxicity. PMID- 10460082 TI - Strychnine poisoning: an uncommon intoxication in children. PMID- 10460083 TI - Olanzapine overdose in a 1-year-old male. PMID- 10460084 TI - Exchange transfusion treatment in a newborn with phenobarbital intoxication. AB - A comatose, 14-day-old boy was referred to our emergency department (ED) after an overdose of phenobarbital, which was used for the treatment of long-standing jaundice. Plasma phenobarbital concentration was 112.4 microg/ml before treatment. One hour after giving albium transfusion, an exchange transfusion, which took about 45 minutes, was performed. Volume of exchange was 400 ml (volume of exchange (ml) = 2 x 85 ml/kg). After the exchange transfusion, the phenobarbital concentration decreased to 50.84 microg/ml. At clinical and laboratory follow-up, the patient recovered fully. This case suggests that exchange transfusion is an effective and successful treatment for phenobarbital intoxication in newborn. PMID- 10460085 TI - Imitation appendicitis: primary omental torsion. PMID- 10460086 TI - Cardiac arrest induced by blunt trauma in children. AB - BACKGROUND: There is incomplete knowledge regarding the outcome of children who suffer a cardiac arrest after blunt trauma. We sought to determine mechanisms of injury, mortality, and rate of organ donation in this population of children. METHODS: Since 1984, all traumatically injured children in San Diego County, California, have been treated at San Diego Children's Hospital. This review encompasses 10,979 pediatric trauma patients evaluated from August 1, 1984 through September 30, 1996. All patients who did not meet the following two criteria were eliminated from the review: 1) a mechanism of blunt trauma, and 2) cardiopulmonary resuscitation performed by a trained medical provider prior to arriving or on arrival to the hospital. A chart review of this set of patients was undertaken to determine mechanism of injury, severity of injury, mortality, and rate of organ donation. RESULTS: In this large metropolitan county, 65 children suffered cardiac arrest following blunt trauma. Accidents involving motor vehicles were the mechanisms responsible for 80% of these injuries. The average Injury Severity Score was 50.3. Mortality was largely related to severe head injury as manifested by a mean Abbreviated Injury Score for head and neck equal to 5.9. All but one of these patients died despite resuscitation. Ninety four percent of these children died within the first 24 hours of injury. The single survivor was discharged in a vegetative state. Solid organs were obtained from 9% of the patients. CONCLUSION: The outcome from blunt cardiac arrest in children is rapidly and nearly uniformly fatal despite resuscitation. Because severe head injuries resulting in brain death are the leading cause of mortality, a significant percentage of organ donations are obtained from these patients. PMID- 10460087 TI - Evaluation and classification of pediatric ocular trauma. AB - PURPOSE: To offer to the pediatric emergency physician consistent and unambiguous terms for the description of pediatric ocular trauma, based upon an adapted version of a standardized classification system. To show the potential effect of this reclassification system in a tertiary care emergency department. METHODS: The authors reviewed a new classification system of ocular trauma and adapted it for use by pediatric emergency physicians. In addition, a retrospective analysis of the records of pediatric patients presenting over a 2-year period to a tertiary emergency department with ocular complaints was performed. The diagnoses related to ocular trauma were reclassified according to the new classification system. RESULTS: Over a 2-year period, 117 pediatric patients were evaluated for ophthalmic complaints. Sixty-seven (57%) of these cases involved an ocular contusion or ruptured globe; however, six disparate diagnoses were given. The cases were reclassified into an adapted, unambiguous, classification system. In some cases, the reclassification altered the indication for immediate ophthalmologic referral. CONCLUSION: There is currently no standardized system of terminology to describe pediatric ocular trauma. This may lead to confusion in communication among the pediatric emergency physician, the pediatrician, and the ophthalmologist. Consistent, unambiguous, terminology will assist in this communication, facilitate the writing of peer-reviewed articles and case reports, and increase the level of accurate documentation in the medical record. PMID- 10460088 TI - Fever, cough, and irritability. PMID- 10460089 TI - Pediatric emergency medicine: legal briefs. PMID- 10460090 TI - Strychnine poisoning. PMID- 10460091 TI - Case records of the Children's Mercy Hospital, Case 02-1999: a 1-month-old infant with respiratory distress and shock. PMID- 10460092 TI - What you see is not always what you get (or want). PMID- 10460093 TI - Proceedings of the 1999 Annual Meeting of the American Academy of Pediatrics, Section of Emergency Medicine. PMID- 10460094 TI - Methamphetamine toxicity. PMID- 10460095 TI - Management of MDR-TB in resource-poor countries. PMID- 10460096 TI - Tuberculosis preventive therapy in HIV-infected persons: feasibility issues in developing countries. AB - There is now strong evidence from several randomized controlled trials for the efficacy of preventive therapy in the prevention of tuberculosis in tuberculin skin test positive persons infected with the human immunodeficiency virus (HIV). The World Health Organization and the International Union Against Tuberculosis and Lung Disease recommend preventive therapy for tuberculin skin test positive, HIV-infected persons who do not have active tuberculosis. While implementation of preventive therapy is manageable in industrialised countries because it is affordable and the infrastructure is in place to screen, treat and monitor patients on a regular basis, its implementation in developing countries presents several problems. Feasibility issues such as identification of large numbers of HIV-infected persons, exclusion of active tuberculosis, identification of those most likely to benefit, supervision of preventive therapy and monitoring of adverse drug reactions need to be resolved before tuberculosis preventive therapy can be introduced on a larger scale in developing countries. Possible sites for implementation of a tuberculosis preventive therapy service include voluntary counselling and testing centres for HIV and occupational health clinics for military personnel, hospital or company workers. Feasibility studies need to be carried out to address these issues in developing countries. PMID- 10460097 TI - Tuberculosis treatment for the beginning of the next century. AB - As we move into the next century it appears that new antituberculosis drugs will arise from four categories: 1) new use of old drugs, 2) new delivery of old drugs, 3) new drugs within old classes, and 4) new classes of drugs. Old drugs such as clofazimine and its analogues, rifabutin, the macrolides, aminoglycosides, quinolones and perhaps vitamin D may find a way into better regimens. New therapy may also arise from new combinations and new uses of current antituberculosis drugs. New drugs are being developed in the rifamycin, fluoroquinolone, and nitroimidazole families. Several immune amplifiers, such as interferon-gamma (IFN-gamma), interleukin-2 (IL-2), and interleukin-12 (IL-12) have undergone pilot testing. Counteracting adhesion molecules is being tested for several infectious diseases. With the unraveling of the tuberculosis genome, attacking enzymes unique to Mycobacterium tuberculosis is easier and allows us to hit elements in both a metabolic pathway and its alternate pathway. Interfering with transcription factors that bind DNA but do not promote RNA production could interrupt transcription. Genetic products of mycobacteria can be modified to cause their own death. Phages may deliver antisense nucleic acids for inhibition of mycobacterial gene expression. The distinction between drugs, immunotherapies and vaccines may blur. PMID- 10460098 TI - Tuberculosis prevention and control activities in the United States: an overview of the organization of tuberculosis services. AB - After a 20% increase in tuberculosis (TB) cases between 1986 and 1992, TB cases in the United States have declined from 1993 through 1997, an average of 5 to 7 per cent per year. In this paper, we review trends and the current epidemiology of TB in the US, present a brief history of TB control efforts in the country, and present the key strategies for TB control in the US. We describe the current organizational structure of TB services in the US, the role of the private sector in TB control, and how TB control is funded. Finally we discuss the mechanisms by which TB policy is developed. The US model represents a categorical disease program that combines a centralized role of the national government in development of policy, funding, and in the maintenance of national surveillance, and a decentralized role of state and local jurisdictions, which adapt and implement national guidelines and which are responsible for day-to-day program activities. Given the relative success of this combined approach, other countries facing the challenge of maintaining an effective TB control program in the face of increased decentralization of health services may find this description useful. PMID- 10460099 TI - A study of maternal mortality at the University Teaching Hospital, Lusaka, Zambia: the emergence of tuberculosis as a major non-obstetric cause of maternal death. AB - SETTING: The Department of Obstetrics and Gynaecology, University Teaching Hospital, Lusaka, Zambia. OBJECTIVES: To ascertain 1) the non-obstetric causes of maternal mortality, 2) the importance of tuberculosis as a cause of maternal deaths, and 3) the trends in the aetiology of non-obstetric causes of maternal deaths during the past decade in the light of the human immunodeficiency virus epidemic. DESIGN: A 2-year retrospective study of the aetiology of all maternal deaths occurring at the University Teaching Hospital (UTH), Lusaka, Zambia between 1 January 1996 and 31 December 1997. Comparison of these data with available data published between 1974 and 1989. RESULTS: There were 251 maternal deaths recorded during the study period. Of these, 106 (42%) were due to direct (obstetric) causes and 145 (58%) were due to indirect (non-obstetric) causes. Malaria (30%), tuberculosis (25%) and unspecified chronic respiratory tract infections (22%) accounted for 77% of non-obstetric causes of maternal deaths and 44% of all causes of maternal deaths. The diagnosis of AIDS was closely linked with that of tuberculosis (92% of cases), and unspecified chronic respiratory illnesses (97%), but not with malaria (37%). The maternal mortality ratio for UTH was calculated at 921 per 100,000 live births, a significant increase from the 118 noted in 1982 and 667 in 1989. CONCLUSIONS: Despite improved obstetric services, the maternal mortality ratios at UTH, Lusaka, have increased eight-fold over the past two decades. This dramatic increase is mainly due to non-obstetric causes of death. Malaria and AIDS-associated tuberculosis and unspecified 'chronic respiratory illnesses' are now major causes of maternal death in Zambia. Greater emphasis is urgently required on early detection, accurate diagnosis, treatment and prevention of malaria and tuberculosis in pregnancy. Further definition of chronic 'unspecified' respiratory illnesses is also required. PMID- 10460100 TI - The effect of immunodeficiency on cutaneous delayed-type hypersensitivity testing in HIV-infected women without anergy: implications for tuberculin testing. HER Study Group. HIV Epidemiology Research. AB - SETTING: A collaborative study in four urban medical centers in the United States. OBJECTIVE: To determine the effect of human immunodeficiency virus (HIV) infection and immunodeficiency on delayed type hypersensitivity (DTH) responses and the implications for interpretation of tuberculin reactions in non-anergic women with or at risk for HIV infection. DESIGN: Demographic and behavioral information, HIV antibody testing, CD4+ lymphocyte counts, and cutaneous responses to DTH testing with mumps, Candida, tetanus toxoid, and tuberculin (purified protein derivative-PPD) antigens were obtained in 1184 women. RESULTS: Reactions to one or more of the four antigens occurred in 436 HIV-seropositive and 356 high-risk seronegative women. Among non-anergic women, HIV-seropositives were less likely (P < or = 0.05) to react to mumps (62% vs 81%), tetanus (72% vs 84%), and PPD (13% vs 19%). Induration in HIV-seropositive reactors was associated with CD4+ cell level for mumps (P = 0.004) and tetanus (P < 0.001), but not for Candida or PPD. HIV-seropositive reactors with CD4+ cell counts >500/mm3 did not have significantly smaller reactions than HIV-seronegatives for any antigen tested. PPD sizes were similar among HIV-seropositive reactors with CD4+ cell counts >500/mm3 (12.4 +/- 7.4 mm) and HIV-seronegative reactors (12.0 +/- 8.3 mm); induration > or =10 mm was seen in 16/173 (9.2%) seropositive women with CD4+ cell counts >500/mm3 and 41/356 (11.5%) seronegative women, respectively (P = 0.5). CONCLUSION: Among HIV-infected women able to react to a DTH antigen, induration in response to that antigen was relatively intact at CD4+ counts >500/mm3. This suggests that degree of immunodeficiency should be considered when interpreting PPD reactions in HIV-infected persons. PMID- 10460101 TI - Drug-resistant pulmonary tuberculosis in Israel, a society of immigrants: 1985 1994. AB - SETTING AND OBJECTIVES: Drug-resistant tuberculosis was uncommon in Israel until 1985, when the waves of immigration began. We studied the incidence and clinical course of resistant pulmonary tuberculosis nationwide. DESIGN: Isolates of Mycobacterium tuberculosis between 1985 and 1994 were surveyed. Data on 150 patients with resistance and 110 patients with drug-sensitive disease were reviewed. Ethnic origin, type of resistance, radiological findings and outcome were analysed. RESULTS: In total, 16.7% of the isolates showed resistance to at least one drug; 58% had resistance to multiple drugs. In 67% of the patients the resistance was primary. Most patients were immigrants from the former USSR and from Ethiopia; none were Israeli-born Jews. Mortality with resistance was 10%, and was highest (14%) with multiple drug resistance. Mortality among drug resistant cases was lowest (3%) among Ethiopian Jews. Cavities and extensive disease were more common with drug resistance. CONCLUSION: Drug resistance has become relatively common in Israel due to immigration from the former USSR and Ethiopia. It is more extensive radiologically and carries a poorer outcome. PMID- 10460102 TI - Treatment of bacillary pulmonary tuberculosis at the chest clinics in the private sector in Korea, 1993. AB - SETTING: Cohort study of bacillary pulmonary tuberculosis patients treated at private sector chest clinics in Korea. OBJECTIVE: To assess the treatment behaviour of physicians in private chest clinics and the treatment outcomes of their patients. DESIGN: 1) A retrospective analysis of a cohort of patients admitted from July through October in 1993, and 2) comparison with results from health centres under the National Tuberculosis Programme (NTP). RESULTS: Nine hundred and sixty bacillary patients (507 newly diagnosed--'new', and 453 retreatment--'old') were admitted to the study. Initial smears and cultures were not performed in 7% and 21%, and follow-up smears and cultures not done in 19% and 28%, respectively. The regimens prescribed were variable: 23 in 'new' and 72 in 'old' patients, 86 in total. Six-month short-course treatment using HRZE was prescribed for 26.2% of 'new' patients. In many instances, the planned treatment duration was excessive. The success rates (cured plus completed) for 'new' and 'old' patients were 74% and 51%, respectively. The failure rates were less than 1% in 'new' and 9% in 'old' patients. CONCLUSION: Prescribed regimens were variable in terms of drug combinations and treatment duration. Overall treatment outcome was inferior to that of the health centres under the NTP. PMID- 10460103 TI - Pharmacokinetics of isoniazid under fasting conditions, with food, and with antacids. AB - STUDY OBJECTIVES: To determine the intra- and intersubject variability in and the effects of food or antacids on the pharmacokinetics of isoniazid (INH). DESIGN: Randomized, four-period cross-over Phase I study in 14 healthy male and female volunteers. Subjects ingested single doses of INH 300 mg under fasting conditions twice, with a high-fat meal, and with aluminum-magnesium antacid. They also received standard doses of rifampin, pyrazinamide, and ethambutol. RESULTS: Serum was collected for 48 hours, and assayed by high performance liquid chromatography (HPLC). Data were analyzed using noncompartmental methods and a compartmental analysis using nonparametric expectation maximization. Both fasting conditions produced similar results: a mean INH Cmax of 5.53 +/- 2.92 microg/ml, Tmax of 1.02 +/- 1.10 hours, and AUC0-infinity of 20.16 +/- 12.45 microg x hr/ml. These findings are similar to those reported previously. Antacids did not alter these parameters significantly (Cmax of 5.62 +/- 2.53 microg/ml, Tmax of 0.71 +/- 0.56 hours, and AUC0-infinity of 20.27 +/- 11.39 microg x hr/ml). In contrast, the high-fat meal recommended by the Food and Drug Administration reduced INH Cmax by 51% (2.73 +/- 1.70 microg/ml), nearly doubled Tmax (1.93 +/- 1.61 hours), and reduced AUC0-infinity by 12% (17.72 +/- 10.32 microg x hr/ml). CONCLUSIONS: These changes in Cmax, Tmax, and AUC0-infinity can be avoided by giving INH on an empty stomach whenever possible. PMID- 10460104 TI - A national survey of human Mycobacterium bovis infection in France. Network of Microbiology Laboratories in France. AB - OBJECTIVE: To evaluate the role of Mycobacterium bovis in the epidemiology of human tuberculosis in France. DESIGN: A national survey in France in 1995 using a questionnaire mailed to all French microbiological laboratories performing mycobacteria cultures. RESULTS: M. bovis was isolated in 38 out of 7075 cases of bacteriologically confirmed tuberculosis (0.5%) notified to the National Reference Centre (CNR) in 1995, resulting in an incidence of 0.07 per 100,000 population. Incidence rates increased with age, and were the highest among patients of 75 years or more (range 0.02-0.33/100,000). Two cases of tuberculosis due to M. bovis were reported in foreign-born children who had come to France for treatment of their disease. No cases were reported among French-born children. The site of tuberculosis was pulmonary in 17 cases, extra-pulmonary in 14, both pulmonary and extra-pulmonary in one, and unknown in six. Extra-pulmonary sites were more frequent in older patients, and pulmonary sites more frequent in younger patients. Two patients were coinfected with the human immunodeficiency virus. Occupational exposure was identified in 13 cases and ingestion of non pasteurised milk in three. In addition, 11 patients had a possible risk of exposure related to their country of birth, family contact or occupation. CONCLUSION: In France, the 0.5% proportion of human tuberculosis due to M. bovis is similar to that of other developed countries. The higher incidence of the disease among older people is likely to reflect the efficacy of the control measures for tuberculosis in cattle. PMID- 10460105 TI - Tuberculosis due to Mycobacterium bovis in the Australian population: cases recorded during 1970-1994. AB - OBJECTIVE: To determine the contribution of Mycobacterium bovis to active tuberculosis in the Australian population during 1970-1994, and to collate and analyse demographic data from bacteriologically proven cases. DESIGN: Summary data for tuberculosis cases notified by Australian public health agencies during 1970-1985 and 1991-1994 were obtained from the database of notifiable diseases maintained by the Department of Health and Family Services. More detailed demographic data for cases confirmed by bacteriology during 1970-1994 were supplied by the Australian Mycobacterium Reference Laboratory Network. RESULTS: At least 236 cases of bovine tuberculosis (TB) occurred in the Australian population during 1970-1994 (mean 9.4 cases; range 3-22 cases annually). The bovine strain has accounted for around 1% of Australian cases of TB during this period. Laboratory sources provided demographic data for 150 cases with positive bacteriology. For this group, the mean age was 54 years (range 22-86), and the male:female ratio was 2.4:1. The majority of cases (74%) involved pulmonary disease. Australian-born persons accounted for 68% of the total cases and typically had histories of employment in meat and/or livestock industries. CONCLUSION: M. bovis was responsible for less than 1.5% of cases of TB in the Australian population during 1970-1994. Most cases were apparently due to reactivation of infection acquired through occupational exposure. Thus, although virtual eradication of M. bovis from Australia's cattle herds has now reduced the risk of exposure, it can be expected that human cases of bovine TB will continue to be detected for years to come. The bovine strain should be considered as the possible agent of TB in foreign-born Australians. PMID- 10460106 TI - Tuberculosis due to Mycobacterium bovis in the Australian population: DNA typing of isolates, 1970-1994. AB - SETTING: Bacteriologically confirmed cases of Mycobacterium bovis in the Australian population. OBJECTIVE: To evaluate the DNA fingerprinting techniques commonly used for M. bovis on isolates from humans and determine whether they were useful for determining the origin of human infection. DESIGN: M. bovis strains isolated between 1970 and 1994 were obtained from five Australian Reference Laboratories. Four DNA fingerprinting techniques, comprising Southern hybridisation with three different probes (the insertion sequence [IS]6110, the polymorphic guanine-cytosine-rich sequence [PGRS] and the direct repeat [DR]) and a PCR-based method (spoligotyping) were used. RESULTS: The PGRS, DR and IS6110 RFLP methods identified 32, 22 and 14 different types respectively from the 45 isolates available. Spoligotyping identified 18 different types. When all methods were combined 41 different strains were identified. Clear differences were found between many isolates from Australian-born patients and those from patients born overseas. CONCLUSIONS: The PGRS RFLP method was the most effective method for typing the human strains, but a combination of methods is recommended for maximum sensitivity. Most Australian-born patients that had worked in the meat and livestock industries were infected with strains similar to those that are commonly found in Australian cattle, confirming the occupational risk in these industries. Patients born overseas were typically infected with strains genetically different from those of patients born in Australia. This suggests that patients born overseas identified with M. bovis were presenting with reactivation of infection acquired outside Australia. PMID- 10460107 TI - Evaluation of a serologic test for the diagnosis of tuberculosis. AB - The ICT TB test, a new, simple, serologic diagnostic test for tuberculosis (TB), was performed on serum samples from individuals seen at an urban teaching hospital and a local health department clinic. The study population included cases of TB, disease with mycobacteria other than tuberculosis (MOTT), non mycobacterial pulmonary disease, and healthy controls. In contrast to prior studies, we found the ICT TB test had little value in detection of new cases of TB (overall sensitivity was 20%). It had very low sensitivity (4%) in the first month of disease. The sensitivity improved in patients tested at least 3 months after clinical presentation, but still remained fairly low. The test was also positive in 30% cases of disease caused by MOTT demonstrating cross-reactivity. It was negative in all human immunodeficiency virus (HIV) positive cases of TB or MOTT. The overall specificity was 89%. At least part of the discrepancy between our results and those of previous investigators may be attributable to differences in the respective study populations, including incidence of HIV disease and duration of tuberculosis illness prior to testing. PMID- 10460108 TI - Sensitivity and specificity of a multi-antigen ELISA test for the serological diagnosis of tuberculosis. AB - A new serological assay (DETECT-TB, BioChem ImmunoSystems), using three recombinant proteins and two synthetic peptides for the detection of the anti Mycobacterium tuberculosis IgG, was evaluated using a panel of serum specimens collected from 100 tuberculosis (TB) patients and 270 controls, in comparison with a homemade ELISA test using purified protein derivative (PPD) as antigen. DETECT-TB presented a higher sensitivity (75%) compared to PPD-ELISA (56%; P < 0.01), while the specificity of each assay was similar (DETECT-TB 97%; PPD-ELISA 100%; P > 0.80). Receiver operating characteristics (ROC) analysis obtained with these data confirmed the higher level of performance of DETECT-TB in comparison with PPD-ELISA. Considering the rapidity, cost-effectiveness and simplicity of this assay, its use may provide useful clinical information aiding in the rapid diagnosis of difficult TB cases. PMID- 10460109 TI - Jabberwocky. PMID- 10460110 TI - HIV infection among patients with tuberculosis in Kenya. PMID- 10460111 TI - The case of drug-resistant pulmonary tuberculosis was nocardiosis! PMID- 10460112 TI - Respiratory manifestations of salmonelloses in AIDS patients. PMID- 10460113 TI - Evaluation of a serological method (ELISA) for the diagnosis of pulmonary tuberculosis in children. PMID- 10460114 TI - Outcome measurement in brain injury rehabilitation--towards a common language. PMID- 10460115 TI - The UK FIM+FAM: development and evaluation. Functional Assessment Measure. AB - BACKGROUND AND AIMS: The aim of this study was to develop and evaluate the UK version of the Functional Assessment Measure (UK FIM+FAM). DESIGN: Before and after evaluation of inter-rater reliability. DEVELOPMENT: Ten 'troublesome' items in the original FIM+FAM were identified as being particularly difficult to score reliably. Revised decision trees were developed and tested for these items over a period of two years to produce the UK FIM+FAM. EVALUATION: A multicentre study was undertaken to test agreement between raters for the UK FIM+FAM, in comparison with the original version, by assessing accuracy of scoring for standard vignettes. METHODS: Baseline testing of the original FIM+FAM was undertaken at the start of the project in 1995. Thirty-seven rehabilitation professionals (11 teams) each rated the same three sets of vignettes - first individually and then as part of a multidisciplinary team. Accuracy was assessed in relation to the agreed 'correct' answers, both for individual and for team scores. Following development of the UK version, the same vignettes (with minimal adaptation to place them in context with the revised version) were rated by 28 individuals (nine teams). RESULTS: Taking all 30 items together, the accuracy for scoring by individuals improved from 74.7% to 77.1% with the UK version, and team scores improved from 83.7% to 86.5%. When the 10 troublesome items were taken together, accuracy of individual raters improved from 69.5% to 74.6% with the UK version (p <0.001), and team scores improved from 78.2% to 84.1% (N/S). For both versions, team ratings were significantly more accurate than individual ratings (p <0.01). Kappa values for team scoring of the troublesome items were all above 0.65 in the UK version. CONCLUSION: The UK FIM+FAM compares favourably with the original version for scoring accuracy and ease of use, and is now sufficiently well developed for wider dissemination. PMID- 10460116 TI - Usability of thenar eminence orthoses: report of a comparative study. AB - OBJECTIVE: To determine the difference in the usability of three types of thenar eminence (TE) orthoses. DESIGN: Prospective comparative pre-experimental study with randomized cross-over design. SETTING: Rehabilitation centre. SUBJECTS: Ten patients with osteoarthritis of the carpometacarpal I joint confirmed by X-ray. INTERVENTION: Three types of TE orthoses, made of either supple elastic material, elastic material with a semi-rigid thumb busk or a semi-rigid (polyethylene) material. MAIN OUTCOME MEASUREMENTS: Pain at the thenar eminence, pinch force, hand function, cosmesis, comfort and function of the TE orthosis. RESULTS: The supple elastic TE orthosis scores significantly better than the more rigid types on the subjects 'comfort', 'function' and the Green Test, while the semi-rigid orthosis scores best on 'cosmesis'. The score on the subject 'pain reduction' shows no significant difference between the three types of TE orthoses. The scores obtained correspond with the outcome of structured interviews about the usability of the TE orthoses. CONCLUSION: Eight out of 10 patients prefer the permanent use of a TE orthosis. Six patients chose the supple elastic orthosis and two chose the semi-rigid orthosis. PMID- 10460117 TI - Trunk kinematics in hemiplegic gait and the effect of walking aids. AB - OBJECTIVE: To establish baseline measurements of trunk movements during hemiplegic gait, to assess the relationship between trunk movements and walking ability, and to investigate the effect of walking aids on the trunk movements. METHOD: Twenty subjects with a chronic hemiplegia from a stroke, who could walk independently, were recruited. Lateral and vertical movements of the pelvis, and symmetry of these movements were measured using CODA (a three-dimensional movement analysis system) as the subjects walked at their own pace without an aid. They were also tested as they walked with a stick and a tripod to assess the effect of different walking aids. Mean values for the trunk movements and symmetry were calculated, Pearson's correlations assessed the relationship between each trunk movement and gait velocity (a measure of overall walking ability), and the influence of the different aids was assessed using a one-way repeated measures ANOVA. RESULTS: Lateral displacement was large (mean = 9.9 cm, SD 3.9) and orientated to the sound side, vertical displacement was small (mean = 2.45 cm, SD 1.4). The movements showed marked asymmetry which favoured the hemiplegic side in that there was less movement of, or towards this side. There was a significant relationship (at 5% level) between walking ability and lateral movements (r = 0.6), but not vertical movements (r= 0.41). No significant differences were found with the different aids. CONCLUSION: These results give baseline values for trunk movements during hemiplegic gait and the relationship between the movements and walking ability. The use of walking aid and the type of walking aid did not affect the subjects' trunk movements or walking ability. PMID- 10460118 TI - Unpacking the black box of therapy -- a pilot study to describe occupational therapy and physiotherapy interventions for people with stroke. AB - OBJECTIVE: To describe the components used in the practice of occupational therapy and physiotherapy for people with stroke and to examine variability between services. DESIGN: A time-sampling strategy in which therapists recorded their face-to-face interventions with stroke patients during 12 weeks over a total of 17 months. SETTINGS AND SUBJECTS: Six occupational therapists and seven physiotherapists from four services (three day hospitals and one domiciliary stroke rehabilitation service) recorded interventions with 89 stroke patients recruited to a larger randomized controlled trial. MAIN OUTCOME MEASURES: Frequencies of use of interventions, together with other details about delivery of therapy, were recorded using a data collection booklet and coding system designed by the participating therapists. RESULTS: The median treatment time for a session was 45 minutes. The most frequently recorded components of physiotherapy intervention were 'walking', 'standing balance' and 'upper limb movement pattern', and of occupational therapy 'physical function', 'social and leisure activities' and 'other'. There was variability between the services in terms of median treatment time, use of intervention codes, frequency of treatment sessions, amount of time spent working with assistance and amount of group work. CONCLUSIONS: The findings support the view that occupational therapy and physiotherapy with people with stroke are not homogeneous activities, and vary between therapists and services. Recommendations include further development of the tool, and use of other methodologies to explore the process and nature of stroke rehabilitation. PMID- 10460119 TI - More than a sympathetic ear? A report on the first year of a writer in residence in a unit for young, physically disabled people. AB - OBJECTIVE: To describe a writer in residence scheme and report the views of patients and staff. DESIGN: Qualitative study including interviews with patients, a questionnaire survey of staff, and dialogue with the writer. SETTING: A unit for young people with physical disability. SUBJECTS: A convenience sample of patients admitted to the unit and members of the multidisciplinary rehabilitation team. INTERVENTION: A writer in residence working 15 hours per week in the unit. RESULTS: A small number of patients worked regularly with the writer and this led to the publication of a play, short stories and poems. Patients described wide ranging benefits including improvements in mood state. Staff regarded the writer in residence scheme as enhancing their treatment and care and thought it helpful in addressing their own psychological needs. The writer saw herself as a catalyst 'for the creativity of others'. CONCLUSION: Such schemes have an important role to play in rehabilitation settings. PMID- 10460120 TI - The process of stroke rehabilitation: what happens and why. AB - OBJECTIVE: To assess the process of hospital-based stroke rehabilitation. DESIGN: A criterion-based process audit, combined with surveys of patient and staff opinions and basic outcome measures. SETTING: All wards and units treating stroke patients in a health care district, including an acute and a community National Health Service (NHS) trust. SUBJECTS: Process audit: documented notes of 115 stroke patients admitted over a four-month period. Patient satisfaction survey: 93 surviving stroke patients. Staff opinion survey: Hospital doctors, therapists and nurses treating stroke patients throughout the district. RESULTS: A disappointingly poor level of service. The main shortcomings were poor assessment of impairment (pass rate, 46%), inadequate communication between staff and with patients and carers (pass rate, 43%), and an absence of rehabilitation beyond the basic of activities of daily living and indoor mobility (pass rate for assessment of disability and emotional need, 50%). Thirty-three per cent of patients were dissatisfied with the hospital-based service they received, particularly lack of therapy, information and recovery. The main reasons for these shortcomings were low priority given to stroke patients, lack of time, shortage of staff, and lack of knowledge and awareness of stroke amongst the staff. Rehabilitation units and elderly care tended to perform better than general medical units in areas of assessment of impairment, self-care skills and mobility, and wheelchair provision. CONCLUSIONS: Evidence from previous publications suggests that this service was no worse than in other districts, but this audit methodology, by comprehensively examining many aspects of a service together, is better able to reveal inadequacies. PMID- 10460121 TI - Persisting symptoms and carers' views of outcome after subarachnoid haemorrhage. AB - OBJECTIVE: To report the outcome of aneurysmal subarachnoid haemorrhage (SAH) in terms of subjective symptoms and carers' perspective over a period of two years. DESIGN: Forty-four consecutive patients admitted to a Regional Neurosurgical Unit and who survived aneurysmal SAH were invited for outpatient assessment at 6, 12 and 24 months post haemorrhage. MEASURES: The Glasgow Outcome Scale (GOS) was used to measure global outcome; the Hospital Anxiety and Depression Scale (HAD) to screen for affective symptoms; the Head Injury Symptom Checklist (HISC) to detect symptoms commonly reported after head trauma; and information was collected from a close friend or relative of the patient using the Relative's Questionnaire (RQ). RESULTS: GOS outcome was significantly related to the severity of the original haemorrhage, as classified by the World Federation of Neurological Surgeons (WFNS) Grading Scale, on admission to hospital. However, even in cases where patients had made a good recovery according to the GOS, a variety of problems were frequently reported by patients and relatives, and many of these persisted for the duration of the study. The three most common and persistent symptoms were tiredness (63%, 59% and 59% at 6, 12 and 24 months respectively), memory disturbance (50%, 52% and 56%) and passivity (61%, 47% and 46%). In contrast, the prevalence of disturbed mood, as reported using the HAD, was similar to that of the general population. CONCLUSIONS: Studies of outcome following SAH should address these subtle disturbances, and information pertaining to potential long-term problems should be provided to patients and relevant others. PMID- 10460122 TI - Social reintegration after stroke: the first stages in the development of the Subjective Index of Physical and Social Outcome (SIPSO). AB - OBJECTIVE: To ensure at outset the content validity of a measure (to be developed) of social integration following stroke. DESIGN: Qualitative study, using in-depth interviews with patients at least six months following stroke. SETTING: Community setting, Bristol, UK. SUBJECTS: Thirty survivors of stroke, discharged at least six months previously from a neurological rehabilitation unit in a district general hospital. RESULTS: Patients reported a change, usually for the worse, in their levels of integration following stroke. More specifically they alluded to a decrease in both the quantity and quality of activities, especially with regard to leisure and those activities occurring outside the home. Social interaction was reduced in most patients, in terms of frequency and quality of contact. From the data, six dimensions of change in quality of interaction emerged. Finally, changes in patients' physical and financial environments were reported. Based on the findings, four diagrams were developed, providing a structure for questionnaire development. CONCLUSIONS: The results of this study confirm the findings of previously reported research. An individual's level of social integration can be affected by stroke in a wide variety of areas and along many different dimensions. Data from this study have highlighted the importance of the patient's subjective impression of the quality of both functional and social performance. This has provided the opportunity to develop a questionnaire based on the patient's perspective, rather than that of health professionals. PMID- 10460123 TI - Maximal grip force in chronic stroke subjects and its relationship to global upper extremity function. AB - OBJECTIVES: Previous studies have shown that recovery of recordable grip strength in acute stroke subjects is one of the most sensitive assessments of initial upper limb recovery and a good prognostic factor for latter recovery. The objectives of this study were to test the reliability of maximal voluntary grip force (MVGF) measures and evaluate the relationship between paretic grip strength deficit and paretic upper extremity function in chronic stroke subjects. DESIGN: Over a three-week period, bilateral MVGF was assessed three times with a modified strain gauge dynamometer in 15 chronic stroke subjects and 10 control subjects. The paretic MVGF deficit was expressed in relation to the MVGF of the nonaffected hand. OUTCOME MEASURES: Upper extremity function in stroke subjects was measured using the Fugl-Meyer, the upper extremity performance test for the elderly (TEMPA), Box and Block and finger-to-nose tests. RESULTS: MVGF measures in both groups of subjects demonstrated good reliability (intraclass correlation, ICC >0.86) and low standard error measurements (SEM). The paretic MVGF of the stroke subjects was greatly impaired in comparison to the control subjects. Results of linear and quadratic regressions analyses show that this impairment was significantly correlated (p <0.01) with the performance of the stroke subjects on the four upper extremity function tests. The percentages of variances explained by the MVGF deficit on all four upper extremity tests varied from 62% to 78% for the linear regressions and from 72% to 93% for the quadratic regressions. CONCLUSIONS: These results suggest that the paretic maximal grip strength, normalized with the maximal grip strength on the nonaffected side, appears to be a valuable outcome measure of upper extremity function in chronic stroke subjects. PMID- 10460124 TI - A methodology for studying the effects of various types of prosthetic feet on the biomechanics of trans-femoral amputee gait. AB - This paper reports on a methodology developed for studying the effects of various types of prosthetic feet on the gait of trans-femoral amputees. It is shown that an analysis in three planes of motion of not only the prosthetic, but also the sound limb provides important information on the performance of prosthetic feet. Two male trans-femoral amputees were tested with four different prosthetic feet; the Springlite II, Carbon Copy III, Seattle LightFoot and the Multiflex foot. A detailed analysis of the results of one amputee and a summary of the most important results of a second subject is presented. The tests were carried out at normal (1.16 m s(-1)) and fast (1.56 m s(-1)) walking speeds. Three dimensional gait analysis was carried out to derive the time curves of the joint angles, intersegmental moments and power at the ankle, knee and hip joints at both the prosthetic and sound sides. A higher first peak of the ground reaction force at the sound side with the Seattle LightFoot compared to that with the Springlite II, may be the result of the lower late stance dorsiflexion angle with the former. Compared to the other two feet, the Carbon Copy III and the Springlite II showed higher prosthetic dorsiflexing moments and positive power at late stance, which could assist in the push-off. The 3D intersegmental loads at the ankle and knee can be used as a guide for design and for compilation of standards for testing of lower limb prostheses incorporating flexible feet. PMID- 10460125 TI - Analysis of stress distribution in the alveolar septa of normal and simulated emphysematic lungs. AB - The alveolar septum consists of a skeleton of fine collagen and elastin fibers, which are interlaced with a capillary network. Its mechanical characteristics play an important role in the overall performance of the lung. An alveolar sac model was developed for numerical analysis of the internal stress distribution and septal displacements within the alveoli of both normal and emphysematic saline-filled lungs. A scanning electron micrograph of the parenchyma was digitized to yield a geometric replica of a typical two-dimensional alveolar sac. The stress-strain relationship of the alveolar tissue was adopted from experimental data. The model was solved by using commercial finite-element software for quasi-static loading of alveolar pressure. Investigation of the state of stresses and displacements in a healthy lung simulation yielded values that compared well with experimentally reported data. Alteration of the mechanical characteristics of the alveolar septa to simulate elastin destruction in the emphysematic model induced significant stress concentrations (e.g., at a lung volume of 60% total capacity, tensions at certain parts in an emphysematic lung were up to 6 times higher than those in a normal lung). The combination of highly elevated stress sites together with the cyclic loading of breathing may explain the observed progressive damage to elastin fibers in emphysematic patients. PMID- 10460126 TI - The influence of pedaling rate on bilateral asymmetry in cycling. AB - The objectives of this study were to (1) determine whether bilateral asymmetry in cycling changed systematically with pedaling rate, (2) determine whether the dominant leg as identified by kicking contributed more to average power over a crank cycle than the other leg, and (3) determine whether the dominant leg asymmetry changed systematically with pedaling rate. To achieve these objectives, data were collected from 11 subjects who pedaled at five different pedaling rates ranging from 60 to 120 rpm at a constant workrate of 260 W. Bilateral pedal dynamometers measured two orthogonal force components in the plane of the bicycle. From these measurements, asymmetry was quantified by three dependent variables, the percent differences in average positive power (%AP), average negative power (%AN), and average crank power (%AC). Differences were taken for two cases--with respect to the leg generating the greater total average for each power quantity at 60 rpm disregarding the measure of dominance, and with respect to the dominant leg as determined by kicking. Simple linear regression analyses were performed on these quantities both for the subject sample and for individual subjects. For the subject sample, only the percent difference in average negative power exhibited a significant linear relationship with pedaling rate; as pedaling rate increased, the asymmetry decreased. Although the kicking dominant leg contributed significantly greater average crank power than the non-dominant leg for the subject sample, the non-dominant leg contributed significantly greater average positive power and average negative power than the dominant leg. However, no significant linear relationships for any of these three quantities with pedaling rate were evident for the subject sample because of high variability in asymmetry among the subjects. For example, significant linear relationships existed between pedaling rates and percent difference in total average power per leg for only four of the 11 subjects and the nature of these relationships was different (e.g. positive versus negative slopes). It was concluded that pedaling asymmetry is highly variable among subjects and that individual subjects may exhibit different systematic changes in asymmetry with pedaling rate depending on the quantity of interest. PMID- 10460127 TI - The tensile and stress relaxation responses of human patellar tendon varies with specimen cross-sectional area. AB - In order to provide insight into the mechanical response of the collagen fascicle structures in tendon, a series of constant strain rate and constant displacement, stress relaxation mechanical tests were performed on sequentially sectioned human patellar tendon specimens (protocol 1) and specimens with both small (approximately 1 mm2) and large (approximately 20 mm2) cross-sectional areas (protocol 2). These data described the stress relaxation and constant strain rate tensile responses as a function of cross-sectional area and water content. The experimental data suggested that small portions of tendon exhibit a higher tensile modulus, a slower rate of relaxation and a lower amount of relaxation in comparison to larger specimens from the same location in the same tendon. The decrease in relaxation response and the increase in tensile modulus with decreasing cross-sectional area was nonlinear. These data suggest that there may be structures other than the subfascicle, such as the epitenon and other connective tissue components, which influence the tensile and stress relaxation responses in tendon. PMID- 10460128 TI - Flow patterns in the radiocephalic arteriovenous fistula: an in vitro study. AB - A significant number of late failures of arteriovenous fistulae for haemodialysis access are related to the progression of intimal hyperplasia. Although the aetiology of this process is still unknown, the geometry of the fistula and the local haemodynamics are thought to be contributory factors. An in-vitro study was carried out to investigate the local haemodynamics in a model of a Cimino-Brescia arteriovenous (AV) fistula with a 30 degrees anastomotic angle and vein-to-artery diameter ratio of 1.6. Flow patterns were obtained by planar illumination of micro-particles suspended in the fluid. Steady and pulsatile flow studies were performed over a range of flow conditions corresponding to those recorded in patients. Quantitative measurements of wall shear stress and turbulence were made using laser Doppler anemometry. The flow structures in pulsatile flow were similar to those seen in steady flow with no significant qualitative changes over the cardiac cycle. This was probably the result of the low pulsatility index of the flow waveform in AV fistulae. Turbulence was the dominant feature in the vein, with relative turbulence intensity > 0.5 within 10 mm of the suture line decreasing to a relatively constant value of about 0.10-0.15 between 40 and 70 mm from the suture line. Peak and mean Reynolds shear stress of 15 and 20 N/m2, respectively, were recorded at the suture line. On the floor of the artery, peak values of temporal mean and oscillating wall shear stress of 9.22 and 29.8 N/m2, respectively. In the vein, both mean and oscillating wall shear stress decreased with distance from the anastomosis. PMID- 10460129 TI - A biomechanical model on muscle forces in the transfer of spinal load to the pelvis and legs. AB - Based on musculoskeletal anatomy of the lower back, abdominal wall, pelvis and upper legs, a biomechanical model has been developed on forces in the load transfer through the pelvis. The aim of this model is to obtain a tool for analyzing the relations between forces in muscles, ligaments and joints in the transfer of gravitational and external load from the upper body via the sacroiliac joints to the legs in normal situations and pathology. The study of the relation between muscle coordination patterns and forces in pelvic structures, in particular the sacroiliac joints, is relevant for a better understanding of the aetiology of low back pain and pelvic pain. The model comprises 94 muscle parts, 6 ligaments and 6 joints. It enables the calculation of forces in pelvic structures in various postures. The calculations are based on a linear/non-linear optimization scheme. To gain a better understanding of the function of individual muscles and ligaments, deviant properties of these structures can be preset. The model is validated by comparing calculations with EMG data from the literature. For agonistic muscles, good agreement is found between model calculations and EMG data. Antagonistic muscle activity is underestimated by the model. Imposed activity of modelled antagonistic muscles has a minor effect on the mutual proportions of agonistic muscle activities. Simulation of asymmetric muscle weakness shows higher activity of especially abdominal muscles. PMID- 10460130 TI - A first estimation of prestress in so-called circularly fibered osteonic lamellae. AB - The existence and role of prestress in the various hierarchical structures of long bone are long standing questions. In this study, the prestress and associated strain that exist in a component of human bone microstructure, circularly fibered osteonic lamella, are estimated. Such estimates allow the formulation of hypotheses on prestress formation and lamellar stiffness. Dimensional measurements were obtained for eight fully calcified lamellae. These dimensions, before isolation from the surrounding alternate osteon and after strain relief by isolation and axial sectioning, furnish data upon which a geometric lamellar model is constructed. A material model is based on the most likely hypothesis as to lamellar structure. This geometric-material model allows estimation of the preexisting strain. The largest strains occur in shear circumferential-axial and normal axial strain directions, averaging 0.08 and 0.05, respectively. The geometric-material model expresses prestress in terms of as yet unknown elastic moduli. The average prestress magnitude is the largest in shear circumferential-axial direction, compensating for alternate osteon weakness in this direction. The estimated axial prestress confirms long hypothesized alternate osteon precompression, which impedes fractures of areas of collagen bundles transverse to the osteon axis at low stresses. The results of the model support the formulation of the following biological hypotheses: (a) lamellar prestress occurs at a supra-molecular level, namely through collagen bundles which are themselves likely to be prestressed; (b) collagen bundles oblique to the lamellar axis are responsible for shear prestress; (c) prestress ranges up to 0.11 GPa; and (d) the lamella is less stiff than alternate osteon. PMID- 10460131 TI - On the kinematic modelling and the parameter estimation of the human shoulder. AB - This paper presents some results on the modelling and the corresponding parameter estimation of the human shoulder. This system consists of the clavicle, the scapula, the humerus and the various joints between these bodies and the trunk through the sternum; it will be represented as a succession of a rotational joint between the sternum and the clavicle and a constant distance joint, representing the scapula between, the clavicle and the humerus head. The parameters of this system are the components of the position vectors of the joint characteristic points (the corresponding centres of the rotations). Experimental results are presented as well as a validation of the proposed model. PMID- 10460132 TI - An improved method for measuring tibiofemoral contact areas in total knee arthroplasty: a comparison of K-scan sensor and Fuji film. AB - A computerised, real time, thin-film pressure transducer method is used to measure tibiofemoral contact area in total knee arthroplasty (TKA) devices that is easier and more reliable and reproducible as compared to the Fuji pressure sensitive film technique. Many authors have suggested that contact areas and pressures within TKA devices can be a predictor of wear and failure of the ultra high molecular weight polyethylene (UHMWPE) tibial insert. In this study, two contact area measurement techniques (Fuji pressure-sensitive Film and K-scan sensor system) were compared using a custom TKA testing jig designed for freedom of movement so that in any loading configuration the component found and seated in its own "home" position. The K-scan system was used to measure contact areas of one TKA design at several angles from 0 to 110 degrees flexion with loads equating to 4, 4.5, and 5 times body weight. For comparison, four ranges of Fuji film were used to measure areas at the same flexion angles but at 5 times body weight only. Contact areas measured with the Fuji films were 11-36% (p < 0.05) lower than those measured by the K-scan sensor. PMID- 10460133 TI - Comparison of physiological and simple pulsatile flows through stenosed arteries. AB - Most experimental and numerical studies of pulsatile flow through stenosed arteries have been performed for a first harmonic oscillatory flow. In this paper, numerical solutions are presented for a physiological pulsatile flow as well as for an equivalent simple pulsatile flow, having the same stroke volume as the physiological flow, and the differences in their flow behavior are discussed. The analysis is restricted to laminar flow, Newtonian fluid and axisymmetric rigid stenosis. Comparison of results shows that the behaviors of the two flows are similar at some instances of time, however, important observed differences indicate that for thorough understanding of pulsatile flow behavior in stenosed arteries, the actual physiological flow should be simulated. PMID- 10460134 TI - Simulation of initial frontside and backside wear rates in a modular acetabular component with multiple screw holes. AB - A sliding distance-based finite element formulation was implemented to predict initial wear rates at the front and back surfaces of a commercially available modular polyethylene component during in vitro loading conditions. We found that contact area, contact stress, and wear at the back surface were more sensitive to the liner/shell conformity than the presence of multiple screw holes. Furthermore, backside linear and volumetric wear rates were at least three orders of magnitude less than respective wear estimates at the articulating surface. This discrepancy was primarily attributed to the difference in maximum sliding distances at the articulating surfaces (measured in mm) versus the back surface (measured in microm). This is the first study in which backside wear has been quantified and explicitly compared with frontside wear using clinically relevant metrics established for the articulating surface. The results of this study suggest that with a polished metal shell, the presence of screw holes does not substantially increase abrasive backside wear when compared with the effects of backside nonconformity. PMID- 10460135 TI - Motion analysis of an articulated locomotion model by video and telemetric data. AB - Traditional techniques of human motion analysis use markers located on body articulations. The position of each marker is extracted from each image. Temporal and kinematic analysis is given by matching these data with a reference model of the human body. However, as human skin is not rigidly linked with the skeleton, each movement causes displacements of the markers and induces uncertainty in results. Moreover, the experiments are mostly conducted in restricted laboratory conditions. The aim of our project was to develop a new method for human motion analysis which needs non-sophisticated recording devices, avoids constraints to the subject studied, and can be used in various surroundings such as stadiums or gymnasiums. Our approach consisted of identifying and locating body parts in image, without markers, by using a multi-sensory sensor. This sensor exploits both data given by a video camera delivering intensity images, and data given by a 3D sensor delivering in-depth images. Our goal, in this design, was to show up the feasibility of our approach. In any case the hardware we used could facilitate an automated motion analysis. We used a linked segment model which referred to Winter's model, and we applied our method not on a human subject but on a life size articulated locomotion model. Our approach consists of finding the posture of this articulated locomotion model in the image. By performing a telemetric image segmentation, we obtained an approximate correspondence between linked segment model position and locomotion model position. This posture was then improved by injecting segmentation results in an intensity image segmentation algorithm. Several tests were conducted with video/telemetric images taken in an outdoor surrounding with the articulated model. This real life-size model was equipped with movable joints which, in static positions, described two strides of a runner. With our fusion method, we obtained relevant limbs identification and location for most postures. PMID- 10460136 TI - A method for synchronising digitised video data. AB - This paper presents a general method for synchronising digitised video data using a mathematical approach based upon the direct linear transformation reconstruction technique. The method was tested using digitised data from genlocked video recordings of gymnastic vaulting, tumbling, high bar and rings. The mean synchronisation error was less than 0.002 s for vaulting and less than 0.001 s for the other activities. PMID- 10460137 TI - A note on an asymptotic solution for the contact of two biphasic cartilage layers in a loaded synovial joint at rest. AB - Retaining the first terms of asymptotic expansions and assuming zero gradient of the contact pressure at the contact edge in the perpendicular direction to the edge, Ateshian et al. (1994, J. Biomechanics 27, 1347-1360; 1992, Adv. Bioeng. ASME 22, 191-194) have presented an asymptotic axially symmetric and plane strain solutions to the contact problems of the two thin biphasic cartilage layers in the synovial joint (with the synovial fluid film in between already depleted), subjected to a sudden normal load. Both the immediate and an early time response to a step loading have been analysed. The present note shows that the contact width thus obtained immediately after the load application differs from the numerical values for a dry frictionless contact of incompressible single-phase elastic layers available in the literature. The difference increases with the decreasing contact width-to-layer thickness ratio. It is proposed to improve the above solutions by releasing zero pressure gradient condition at the contact edge and calculating the instantaneous contact widths from the equations proposed for the corresponding case by Matthewson (1981, J. Mech. Phys. Solids 29, 89-113) and Meijers (1967, Appl. Sci. Res. 18, 353-383) that approximate better the numerical values. The instantaneous rate of change in the contact width is then obtained for a step load varying in time. PMID- 10460138 TI - Active responses decrease impact forces at the hip and shoulder in falls to the side. AB - Active responses, such as using the arm to break the fall, may be an effective means of decreasing likelihood of injury in a fall and may help explain why only a small percentage of falls result in a fracture. We quantified the impact force at the hip and shoulder in falls to the side from a kneeling position under three conditions: (1) attempting to break the fall by using an arm; (2) falling with the body relaxed; and (3) falling with the body tensed. Subjects fell from a kneeling position onto a force platform array covered with foam padding and impact force data were recorded. The ground reaction force-time curve was generally bimodal due to sequential impacts of the hip and shoulder. Impact forces at the hip and shoulder were 12 and 16% less for the slap condition (p < 0.05) than for the tensed condition. The impact forces for the relaxed and tensed conditions were not significantly different, although impact forces tended to be less in the relaxed condition. We concluded that active responses reduce the impact forces experienced at the hip and shoulder in falls to the side. Decreased effectiveness of protective responses, due to increases in reaction time and decreases in strength with age, may help explain why so many hip fractures occur in the elderly but so few occur in younger people. PMID- 10460139 TI - Mechanical characterization of the bovine iris. AB - Quantification of the mechanical properties of the iris is necessary to assess the clinical significance of passive iris deformation, which has been suggested as a mechanism for certain forms of glaucoma. Extension tests were performed on isolated bovine irises to determine the passive mechanical behavior of the iris and the contribution of each of its two constituent muscles, the sphincter iridis and the dilator pupillae, to the overall properties. Because of the shape of the iris and our desire to use intact tissue, a "loop" experiment was performed in which the iris was stretched by hooking the sample and pulling. A simple mathematical model was used to account for the geometry of the experiment and the progressive recruitment of tissue. Radial extension experiments on samples dissected from the iris were also performed. The iris was found to be anisotropic, elastic, and incompressible. The average azimuthal Young's modulus of the sphincter was found to be 340 kPa; the average azimuthal Young's modulus of the dilator was found to be 890 kPa, which was significantly higher (p < 0.01). The radial Young's modulus of the dilator was found to be 9.6 kPa, much lower than the azimuthal value. PMID- 10460140 TI - Respiratory distress after use of droperidol for agitation. PMID- 10460141 TI - Pathogenesis of envenomation. PMID- 10460142 TI - Medical direction for staffing of ambulances. EMS Committee. PMID- 10460143 TI - Treatment of multiple miliary osteoma cutis with tretinoin gel. PMID- 10460145 TI - Phenylalanine residues in the active site of tyrosine hydroxylase: mutagenesis of Phe300 and Phe309 to alanine and metal ion-catalyzed hydroxylation of Phe300. AB - Residues Phe300 and Phe309 of tyrosine hydroxylase are located in the active site in the recently described three-dimensional structure of the enzyme, where they have been proposed to play roles in substrate binding. Also based on the structure, Phe300 has been reported to be hydroxylated due to a naturally occurring posttranslational modification [Goodwill, K. E., Sabatier, C., and Stevens, R. C. (1998) Biochemistry 37, 13437-13445]. Mutants of tyrosine hydroxylase with alanine substituted for Phe300 or Phe309 have now been purified and characterized. The F309A protein possesses 40% less activity than wild-type tyrosine hydroxylase in the production of DOPA, but full activity in the production of dihydropterin. The F300A protein shows a 2.5-fold decrease in activity in the production of both DOPA and dihydropterin. The K(6-MPH4) value for F300A tyrosine hydroxylase is twice the wild-type value. These results are consistent with Phe309 having a role in maintaining the integrity of the active site, while Phe300 contributes less than 1 kcal/mol to binding tetrahydropterin. Characterization of Phe300 by MALDI-TOF mass spectrometry and amino acid sequencing showed that hydroxylation only occurs in the isolated catalytic domain after incubation with a large excess of 7, 8-dihydropterin, DTT, and Fe(2+). The modification is not observed in the untreated catalytic domain or in the full length protein, even in the presence of excess iron. These results establish that hydroxylation of Phe300 is an artifact of the crystallography conditions and is not relevant to catalysis. PMID- 10460146 TI - Hammerhead ribozyme mechanism: a ribonucleotide 5' to the substrate cleavage site is not essential. AB - Three hammerhead ribozymes with triplet specificities for cleavage 3' of CUC, GUC, and GUA have been evaluated for their sensitivity to the substitution of thymidine or 2'-deoxyuridine at central nucleotide position 16.1 in the substrate triplet. All three ribozymes cleaved their respective substrates, containing uridine or the modifications, with comparable rates. This indicates that the 2' hydroxy group at position 16.1 is not essential for activity even though X-ray structure analysis shows it participates in H-bonding interactions. These H-bonds were considered to be of functional significance because an earlier report had provided data that thymidine at position 16.1 is deleterious for catalytic activity [Yang, J.-H., Perreault, J.-P., Labuda, D., Usman, N., and Cedergren, R. (1990) Biochemistry 29, 11156-11160]. PMID- 10460147 TI - Mechanism of DnaB helicase of Escherichia coli: structural domains involved in ATP hydrolysis, DNA binding, and oligomerization. AB - We describe the delineation of three distinct structural domains of the DnaB helicase of Escherichia coli: domain alpha, amino acid residues (aa) 1-156; domain beta, aa 157-302; and domain gamma, aa 303-471. Using mutants with deletion in these domains, we have examined their role(s) in hexamer formation, DNA-dependent ATPase, and DNA helicase activities. The mutant DnaBbetagamma protein, in which domain alpha was deleted, formed a hexamer; whereas the mutant DnaBalphabeta, in which domain gamma was deleted, could form only dimers. The dimerization of DnaBalphabeta was Mg(2+) dependent. These data suggest that the oligomerization of DnaB helicase involves at least two distinct protein-protein interaction sites; one of these sites is located primarily within domain beta (site 1), while the other interaction site is located within domain gamma (site 2). The mutant DnaBbeta, a polypeptide of 147 aa, where both domains alpha and gamma were deleted, displayed a completely functional ATPase activity. This domain, thus, constitutes the "central catalytic domain" for ATPase activity. The ATPase activity of DnaBalphabeta was kinetically comparable to that of DnaBbeta, indicating that domain alpha had little or no influence on the ATPase activity. In both cases, the ATPase activities were DNA independent. DnaBbetagamma had a DNA-dependent ATPase activity that was kinetically comparable to the ATPase activity of wild-type DnaB protein (wtDnaB), indicating a specific role for C terminal domain gamma in enhancement of the ATPase activity of domain beta as well as in DNA binding. Mutant DnaBbetagamma, which lacked domain alpha, was devoid of any helicase activity pointing to a significant role for domain alpha. The major findings of this study are (i) domain beta contained a functional ATPase active site; (ii) domain gamma appeared to be the DNA binding domain and a positive regulator of the ATPase activity of domain beta; (iii) although domain alpha did not have any significant effect on the ATPase, DNA binding activities, or hexamer formation, it definitely plays a pivotal role in transducing the energy of ATP hydrolysis to DNA unwinding by the hexamer; and (iv) all three domains are required for helicase activity. PMID- 10460148 TI - Mechanism of DNA binding by the DnaB helicase of Escherichia coli: analysis of the roles of domain gamma in DNA binding. AB - We have analyzed the mechanism of single-stranded DNA (ssDNA) binding mediated by the C-terminal domain gamma of the DnaB helicase of Escherichia coli. Sequence analysis of this domain indicated a specific basic region, "RSRARR", and a leucine zipper motif that are likely involved in ssDNA binding. We have carried out deletion as well as in vitro mutagenesis of specific amino acid residues in this region in order to determine their function(s) in DNA binding. The functions of the RSRARR domain in DNA binding were analyzed by site-directed mutagenesis. DnaBMut1, with mutations R(328)A and R(329)A, had a significant decrease in the DNA dependence of ATPase activity and lost its DNA helicase activity completely, indicating the important roles of these residues in DNA binding and helicase activities. DnaBMut2, with mutations R(324)A and R(326)A, had significantly attenuated DNA binding as well as DNA-dependent ATPase and DNA helicase activities, indicating that these residues also play a role in DNA binding and helicase activities. The role(s) of the leucine zipper dimerization motif was (were) determined by deletion analysis. The DnaB Delta 1 mutant with a 55 amino acid C-terminal deletion, which left the leucine zipper and basic DNA binding regions intact, retained DNA binding as well as DNA helicase activities. However, the DnaB Delta 2 mutant with a 113 amino acid C-terminal deletion that included the leucine zipper dimerization motif, but not the RSRARR sequence, lost DNA binding, DNA helicase activities, and hexamer formation. The major findings of this study are (i) the leucine zipper dimerization domain, I(361)-L(389), is absolutely required for (a) dimerization and (b) ssDNA binding; (ii) the base rich RSRARR sequence is required for DNA binding; (iii) three regions of domain gamma (gamma I, gamma II, and gamma III) differentially regulate the ATPase activity; (iv) there are likely three ssDNA binding sites per hexamer; and (v) a working model of DNA unwinding by the DnaB hexamer is proposed. PMID- 10460149 TI - Engineered recombinant factor VII Q217 variants with altered inhibitor specificities. AB - Recombinant factor VII with residue 217 (chymotrypsinogen numbering system) converted to alanine (VIIQ217A), glutamic acid (VIIQ217E), or glycine (VIIQ217G) was characterized. In a prothrombin time assay, VIIQ217E demonstrated 100%, VIIQ217A 15%, and VIIQ217G <1% clotting activities relative to wild-type VII. Binding of VIIQ217A and VIIQ217G to TF was comparable to that of wild-type VII to TF. All the variants were readily activated by factor Xa. Autoactivation in the presence of TF was efficient with VIIQ217E, slow with VIIQ217A, but undetected with VIIQ217G. Relative to wild-type VII added at the same concentration, VIIQ217E had no effect on the PT of normal plasma, whereas VIIQ217A slightly and VIIQ217G dramatically prolonged the clotting time in a dose-dependent manner. Activation of macromolecular substrates paralleled this functional inhibition. The k(cat)/K(M) values for factor X activation in the presence of TF were 2.4 for VIIaQ217E as compared to 1.9 (M(-)(1) s(-)(1) x 10(7)) for wild-type VIIa, 1.57 for VIIaQ217A, and 0.05 with VIIaQ217G. In comparison to wild-type VIIa, VIIaQ217E cleaved the chromogenic substrate S2765 (Z-D-Arg-Gly-Arg-pNA) with 10 fold higher k(cat). Analysis of the interactions with the inhibitors TFPI and antithrombin III demonstrated that VIIaQ217A but not VIIaQ217E or VIIaQ217G was inhibited less efficiently by TFPI either in the presence or in the absence of factor Xa. In contrast, VIIaQ217A association with antithrombin III in the presence of heparin was the fastest among the variants with a second-order rate constant of 2.31 (x10(3) M(-)(1) min(-)(1)), as compared to 0.47 and 1.47 for VIIaQ217E and wild-type VIIa, respectively. Our results demonstrate that residue Q(217) is important in regulating substrate and, more importantly, inhibitor recognition by VIIa. PMID- 10460150 TI - A molecular envelope of the ligand-binding domain of a glutamate receptor in the presence and absence of agonist. AB - Solution scattering studies were performed on a ligand-binding domain (S1S2) of a glutamate receptor ion channel (GluR) in order to study GluR-binding and signal transduction mechanisms. The core of the ligand-binding domain is homologous to prokaryotic periplasmic binding proteins (PBP), whose binding mechanism involves a dramatic cleft closure: the "Venus flytrap". Several models of GluR function have proposed that a similar cleft closure is induced by agonist binding. We have directly tested this putative functional homology by measuring the radius of gyration of S1S2 in the presence and absence of saturating concentrations of agonists. In contrast to the PBP, S1S2 shows no reduction in radius of gyration upon agonist binding, excluding a comparably large conformational change. Furthermore, we determined an ab initio molecular envelope for our S1S2 construct, which also contains the peptides that connect the PBP homology core to the three transmembrane domains and to an N-terminal domain. By fitting an atomic model of the ligand-binding domain core to the envelope of our extended construct, we were able to establish the likely position of these connecting peptides. Their positions relative to one another and to the expected sites of an agonist-induced conformational change suggest that ion channel gating and desensitization may involve more subtle and complex mechanisms than have been assumed based on the structural homology to the PBP. PMID- 10460151 TI - Probing the role of metal ions in RNA catalysis: kinetic and thermodynamic characterization of a metal ion interaction with the 2'-moiety of the guanosine nucleophile in the Tetrahymena group I ribozyme. AB - Deciphering the role of individual metal ions in RNA catalysis is a tremendous challenge, as numerous metal ions coat the charged backbone of a folded RNA. Metal ion specificity switch experiments combined with quantitative analysis may provide a powerful tool for probing specific metal ion-RNA interactions and for delineating the role of individual metal ions among the sea of metal ions bound to RNA. We show herein that Mn(2+) rescues the deleterious effect of replacing the 2'-OH of the guanosine nucleophile (G) by -NH(2) (G(NH)()2) in the reaction catalyzed by the Tetrahymena group I ribozyme (E), and the Mn(2+) concentration dependence suggests that a single metal ion is responsible for rescue. This provides strong evidence for a metal ion interaction with the 2'-moiety of G in this ribozyme (referred to as M(C)), confirming and extending previous results in a bacteriophage group I intron [Sjogren, A.-S., Pettersson, E., Sjoberg, B.-M., and Stromberg, R. (1997) Nucleic Acids Res. 25, 648-654]. Toward understanding the >10(6)-fold catalytic contribution of the 2'-OH of G, we have determined the individual reaction steps affected by M(C) and quantitated these effects. has only a small effect on binding of G(NH)()2 to the free ribozyme or ribozyme.oligonucleotide complexes that lack the reactive phosphoryl group. In contrast, increases the binding of G(NH)()2 to the ribozyme.oligonucleotide substrate (E.S) complex 20-fold and increases the binding of S to the E.G(NH)()2 complex by the same amount. These and other observations suggest that M(C) plays an integral role in the coupled binding of the oligonucleotide substrate and the guanosine nucleophile. This metal ion may be used to align the nucleophile within the active site, thereby facilitating the reaction. Alternatively or in addition, M(C) may act in concert with an additional metal ion to coordinate and activate the 3'-OH of G. Finally, these experiments have also allowed us to probe the properties of this metal ion site and isolate the energetic effects of the interaction of this specific metal ion with the 2'-moiety of G. PMID- 10460152 TI - Protonated 2'-aminoguanosine as a probe of the electrostatic environment of the active site of the Tetrahymena group I ribozyme. AB - We have probed the electrostatic environment of the active site of the Tetrahymena group I ribozyme (E) using protonated 2'-aminoguanosine (), in which the 2'-OH of the guanosine nucleophile (G) is replaced by an group. At low concentrations of divalent metal ion (2 mM Mg(2+)), binds at least 200-fold stronger than G or G(NH)()2, with a dissociation constant of 5' direction. In addition, the DNA adducts of these complexes inhibited DNA transcription in vitro. Thus, the binding of the 1,1/c, c complexes modifies DNA in a way that is distinctly different from the modification by the antitumor drug cisplatin. In addition, there are significant differences between the dinuclear 1,1/c,c and 1,1/t, t isomers. The results of this work are consistent with the hypothesis and support the view that platinum drugs that bind to DNA in a fundamentally different manner can exhibit different biological properties including the spectrum and intensity of antitumor activity. The intracellular DNA binding of the dinuclear compounds is compared to the results presented here. It has been suggested that differences in cross-link structure may be an important factor underlying their different biological efficiencies. PMID- 10460155 TI - Effect of cysteine residues on the activity of arginyl-tRNA synthetase from Escherichia coli. AB - Arginyl-tRNA synthetase (ArgRS) from Escherichia coli (E. coli) contains four cysteine residues. In this study, the role of cysteine residues in the enzyme has been investigated by chemical modification and site-directed mutagenesis. Titration of sulfhydryl groups in ArgRS by 5, 5'-dithiobis(2-nitro benzoic acid) (DTNB) suggested that a disulfide bond was not formed in the enzyme and that, in the native condition, two DTNB-sensitive cysteine residues were located on the surface of ArgRS, while the other two were buried inside. Chemical modification of the native enzyme by iodoacetamide (IAA) affected only one DTNB-sensitive cysteine residue and resulted in 50% loss of enzyme activity, while modification by N-ethylmeimide (NEM) affected two DTNB-sensitive residues and caused a complete loss of activity. These results, when combined with substrate protection experiments, suggested that at least the two cysteine residues located on the surface of the molecule were directly involved in substrates binding and catalysis. However, changing Cys to Ala only resulted in slight loss of enzymatic activity and substrate binding, suggesting that these four cysteine residues in E. coli ArgRS were not essential to the enzymatic activity. Moreover, modifications of the mutant enzymes indicated that the two DTNB- and NEM sensitive residues were Cys(320) and Cys(537) and the IAA-sensitive was Cys(320). Our study suggested that inactivation of E. coli ArgRS by sulfhydryl reagents is a result of steric hindrance in the enzyme. PMID- 10460156 TI - Identification of linker regions and domain borders of the transcription activator protein NtrC from Escherichia coli by limited proteolysis, in-gel digestion, and mass spectrometry. AB - We have developed a mass spectrometry based method for the identification of linker regions and domain borders in multidomain proteins. This approach combines limited proteolysis and in-gel proteolytic digestions and was applied to the determination of linkers in the transcription factor NtrC from Escherichia coli. Limited proteolysis of NtrC with thermolysin and papain revealed that initial digestion yielded two major bands in SDS-PAGE that were identified by mass spectrometry as the R-domain and the still covalently linked OC-domains. Subsequent steps in limited proteolysis afforded further cleavage of the OC fragment into the O- and the C-domain at accessible amino acid residues. Mass spectrometric identification of the tryptic/thermolytic peptides obtained after in-gel total proteolysis of the SDS-PAGE-separated domains determined the domain borders and showed that the protease accessible linker between R- and O-domain comprised amino acids Val-131 and Gln-132 within the "Q-linker" in agreement with papain and subtilisin digestion. The region between amino acid residues Thr-389 and Gln-396 marked the hitherto unknown linker sequence that connects the O- with the C-domain. High abundances of proline-, alanine-, serine-, and glutamic acid residues were found in this linker structure (PASE-linker) of related NtrC response regulator proteins. While R- and C-domains remained stable under the applied limited proteolysis conditions, the O-domain was further truncated yielding a core fragment that comprised the sequence from Ile-140 to Arg-320. ATPase activity was lost after separation of the R-domain from the OC-fragment. However, binding of OC- and C- fragments to specific DNA was observed by characteristic band-shifts in migration retardation assays, indicating intact tertiary structures of the C-domain. The outlined strategy proved to be highly efficient and afforded lead information of tertiary structural features necessary for protein design and engineering and for structure-function studies. PMID- 10460157 TI - Replication protein A stimulates proliferating cell nuclear antigen-dependent repair of abasic sites in DNA by human cell extracts. AB - Base excision repair (BER) pathway is the major cellular process for removal of endogenous base lesions and apurinic/apyrimidinic (AP) sites in DNA. There are two base excision repair subpathways in mammalian cells, characterized by the number of nucleotides synthesized into the excision patch. They are the "single nucleotide" (one nucleotide incorporated) and the "long-patch" (several nucleotides incorporated) BER pathways. Proliferating cell nuclear antigen (PCNA) is known to be an essential factor in long-patch base excision repair. We have studied the role of replication protein A (RPA) in PCNA-dependent, long-patch BER of AP sites in human cell extracts. PCNA and RPA were separated from the other BER proteins by fractionation of human whole-cell extract on a phosphocellulose column. The protein fraction PC-FII (phosphocellulose fraction II), which does not contain RPA and PCNA but otherwise contains all core BER proteins required for PCNA-dependent BER (AP endonuclease, DNA polymerases delta, beta and DNA ligase, and FEN1 endonuclease), had reduced ability to repair plasmid DNA containing AP sites. Purified PCNA or RPA, when added separately, could only partially restore the PC-FII repair activity of AP sites. However, additions of both proteins together greatly stimulated AP site repair by PC-FII. These results demonstrate a role for RPA in PCNA-dependent BER of AP sites. PMID- 10460159 TI - Tertiary structure stabilization promotes hairpin ribozyme ligation. AB - The hairpin ribozyme catalyzes a reversible RNA cleavage reaction that participates in processing intermediates of viral satellite RNA replication in plants. A minimal hairpin ribozyme consists of two helix-loop-helix segments. These segments associate noncoaxially in the active folded structure in a way that brings catalytically important loop nucleotides into close proximity. The hairpin ribozyme in the satellite RNA of Tobacco Ringspot Virus assembles in the context of a four-way helical junction. Recent physical characterization of hairpin ribozyme structures using fluorescence resonance energy transfer demonstrated enhanced stability of the folded structure in the context of a four way helical junction compared to minimal hairpin ribozyme variants. Analysis of the functional consequences of this modification of the helical junction has revealed two changes in the hairpin ribozyme kinetic mechanism. First, ribozymes with a four-way helical junction bind 3' cleavage products with much higher affinity than minimal hairpin ribozymes, evidence that tertiary interactions within the folded structure contribute to product binding energy. Second, the balance between ligation and cleavage shifts in favor of ligation. The enhanced ligation activity of hairpin ribozymes that contain a four-way helical junction supports the notion that tertiary structure stability is a major determinant of the hairpin ribozyme proficiency as a ligase and illustrates the link between RNA structure and biological function. PMID- 10460158 TI - Effect of DNA polymerases and high mobility group protein 1 on the carrier ligand specificity for translesion synthesis past platinum-DNA adducts. AB - Translesion synthesis past Pt-DNA adducts can affect both the cytotoxicity and mutagenicity of the platinum adducts. We have shown previously that the extent of replicative bypass in vivo is influenced by the carrier ligand of platinum adducts. The specificity of replicative bypass may be determined by the DNA polymerase complexes that catalyze translesion synthesis past Pt-DNA adducts and/or by DNA damage-recognition proteins that bind to the Pt-DNA adducts and block translesion replication. In the present study, primer extension on DNA templates containing site-specifically placed cisplatin, oxaliplatin, JM216, or chlorodiethylenetriamine-Pt adducts revealed that the eukaryotic DNA polymerases beta, zeta, gamma, and human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) had a similar specificity for translesion synthesis past Pt-DNA adducts (dien >> oxaliplatin >/= cisplatin > JM216). Primer extension assays performed in the presence of high mobility group protein 1 (HMG1), which is known to recognize cisplatin-damaged DNA, revealed that inhibition of translesion synthesis by HMG1 also depended on the carrier ligand of the Pt-DNA adduct (cisplatin > oxaliplatin = JM216 >> dien). These data were consistent with the results of gel-shift experiments showing similar differences in the affinity of HMG1 for DNA modified with the different platinum adducts. Our studies show that both DNA polymerases and damage-recognition proteins can impart specificity to replicative bypass of Pt-DNA adducts. This information may serve as a model for further studies of translesion synthesis. PMID- 10460160 TI - NMR characterization of substrate binding in the phthalate dioxygenase system. AB - The paramagnetic enhancements in the NMR relaxation rates for the fluorine in fluorophthalates have been used to determine the position of the phthalate with respect to the mononuclear metal ion in native and metal-substituted derivatives of phthalate dioxygenase (PDO). These studies show directly that the substrate interacts with the mononuclear metal of PDO and provide the first structural characterization of this interaction. With a molecular mass of 200 kDa, PDO is one of the largest proteins studied to date by paramagnetic NMR. Two paramagnetically broadened (19)F lines were observed for monofluorophthalates bound to CoPDO. This demonstrates that fluorophthalate binds to PDO with a handedness, i.e., with the fluorine label facing to the "right" or to the "left", relative to the hyperfine tensor of the Co(II). The relative affinities of the two orientations are slightly different, with a 2-fold and 5-fold excess of the preferred orientation for 4-fluorophthalate and 3-fluorophthalate, respectively. The longitudinal relaxation rate (T(1)) and transverse relaxation rate (T(2)) data give mutually consistent fluorine to cobalt distances. These results are consistent with approximate bilateral symmetry, with the Co to 3-fluorophthalate distances ( approximately 5.5 A) approximately 25% longer than the Co to 4 fluorophthalate distances ( approximately 4. 5 A). A detailed geometric model is derived from these data. This structural characterization of the mononuclear site provides a framework to develop hypotheses for the mechanism of oxygenation by the Fe(II)-containing aromatic dioxygenases. PMID- 10460161 TI - Electron paramagnetic resonance measurements of the ferrous mononuclear site of phthalate dioxygenase substituted with alternate divalent metal ions: direct evidence for ligation of two histidines in the copper(II)-reconstituted protein. AB - The metalloenzyme phthalate dioxygenase (PDO) contains two iron-based sites. A Rieske-type [2Fe-2S] cluster serves as an electron-transferring cofactor, and a mononuclear iron site is the putative site of substrate oxygenation. A reductase, which contains FMN and a plant-type [2Fe-2S] ferredoxin domain, transfers electrons from NADH to the Rieske center. Any of the metal ions, Fe(II), Cu(II), Co(II), Mn(II), and Zn(II), can be used to populate the mononuclear site, but only Fe(II) is competent for effecting hydroxylation. Nevertheless, studies of how these metal ions affect both the EPR spectra of the reduced Rieske site and the kinetics of electron transfer in the PDO system indicated that each of these metal ions binds tightly and affects the protein similarly. In this study, EPR spectra were obtained from samples in which iron of the mononuclear site was replaced with Cu(II). The use of (63)Cu(II), in combination with PDO obtained from cultures grown on media enriched in (15)N [using ((15)NH(4))(2)SO(4) as a sole nitrogen source], [delta,epsilon-(15)N]histidine, as well as natural abundance sources of nitrogen, enabled detailed spectral analysis of the superhyperfine structure of the Cu(II) EPR lines. These studies clearly show that two histidines are coordinated to the mononuclear site. Coupled with previous studies [Bertini, I., Luchinat, C., Mincione, G., Parigi, G., Gassner G. T., and Ballou, D. P. (1996) J. Bioinorg. Chem. 1, 468-475] that show the presence of one or two water molecules coordinated to the iron, it is suggested that the mononuclear site is similar to several other mononuclear nonheme iron proteins, including naphthalene dioxygenase, for which crystal structures are available. The lack of observable EPR interaction signals between Cu(II) in the mononuclear site and the reduced Rieske center of PDO suggest that the two sites are at least 12 A apart, which is similar to that found in the naphthalene dioxygenase crystal structure. PMID- 10460163 TI - The methionyl aminopeptidase from Escherichia coli can function as an iron(II) enzyme. AB - The identity of the physiologically relevant metal ions for the methionyl aminopeptidase (MetAP) from Escherichia coli was investigated and is suggested to be Fe(II). The metal content of whole cells in the absence and presence of expression of the type I MetAP from E. coli was determined by inductively coupled plasma (ICP) emission analysis. The observed change in whole cell concentrations of cobalt, cadmium, copper, nickel, strontium, titanium, and vanadium upon expression of MetAP was negligible. On the other hand, significant increases in the cellular metal ion concentrations of chromium, zinc, manganese, and iron were observed with the increase in iron concentration being 4.4 and 6.2 times greater than that of manganese and zinc, respectively. Activity assays of freshly lysed BL21(DE3) cells containing the pMetAAP plasmid revealed detectable levels (>2 units/mg) of MetAP activity. Control experiments with BL21(DE3) without the MetAP plasmid showed no detectable enzymatic activity. Since MetAP is active upon expression, these data strongly suggest that cobalt is not the in vivo metal ion for the MetAP from E. coli. The MetAP from E. coli as purified was found to be catalytically inactive ( 34 kDa > 36 kDa > 38 kDa. The 34 and 38 kDa polypeptides react with an antibody specific for the N-terminus of the M1 isoform, and 32 and 36 kDa correspond to the shorter M23 isoform. Immunogold electron microscopic studies with rat cerebellum cryosections demonstrated that the 34 kDa polypeptide colocalizes in perivascular astrocyte endfeet where the 32 kDa polypeptide is abundantly expressed. Velocity sedimentation, cross-linking, and immunoprecipitation analyses of detergent-solubilized rat brain revealed that the 32 and 34 kDa polypeptides reside within heterotetramers. Immunoprecipitation of AQP4 expressed in Xenopus laevis oocytes demonstrated that heterotetramer formation reflects the relative expression levels of the 32 and 34 kDa polypeptides; however, tetramers containing different compositions of the two polypeptides exhibit similar water permeabilities. These studies demonstrate that AQP4 heterotetramers are formed from two overlapping polypeptides and indicate that the 22-amino acid sequence at the N-terminus of the 34 kDa polypeptide does not influence water permeability but may contribute to membrane trafficking or assembly of arrays. PMID- 10460173 TI - 1H NMR studies on the CuA center of nitrous oxide reductase from Pseudomonas stutzeri. AB - 1H NMR spectra of the CuA center of N2OR from Pseudomonas stutzeri, and a mutant enzyme that contains only CuA, were recorded in both H2O- and D2O-buffered solution at pH 7.5. Several sharp, well-resolved hyperfine-shifted 1H NMR signals were observed in the 60 to -10 ppm chemical shift range. Comparison of the native and mutant N2OR spectra recorded in H2O-buffered solutions indicated that several additional signals are present in the native protein spectrum. These signals are attributed to a dinuclear copperII center. At least two of the observed hyperfine shifted signals associated with the dinuclear center, those at 23.0 and 13.2 ppm, are lost upon replacement of H2O buffer with D2O buffer. These data indicate that at least two histidine residues are ligands of a dinuclear CuII center. Comparison of the mutant N2OR 1H NMR spectra recorded in H2O and D2O indicates that three signals, c (27.5 ppm), e (23.6 ppm), and i (12.4 ppm), are solvent exchangeable. The two most strongly downfield-shifted signals (c and e) are assigned to the two N epsilon 2H (N-H) protons of the coordinated histidine residues, while the remaining exchangeable signal is assigned to a backbone N-H proton in close proximity to the CuA cluster. Signal e was found to decrease in intensity as the temperature was increased, indicating that proton e resides on a more solvent-exposed histidine residue. One-dimensional nOe studies at pH 7.5 allowed the histidine ring protons to be definitively assigned, while the remaining signals were assigned by comparison to previously reported spectra from CuA centers. The temperature dependence of the observed hyperfine-shifted 1H NMR signals of mutant N2OR were recorded over the temperature range of 276-315 K. Both Curie and anti-Curie temperature dependencies are observed for sets of hyperfine-shifted protons. Signals a and h (cysteine protons) follow anti-Curie behavior (contact shift increases with increasing temperatures), while signals b g, i, and j (histidine protons) follow Curie behavior (contact shift decreases with increasing temperatures). Fits of the temperature dependence of the observed hyperfine-shifted signals provided the energy separation (Delta EL) between the ground (2B3u) and excited (2B2u) states. The temperature data obtained for all of the observed hyperfine-shifted histidine ligand protons provided a Delta EL value of 62 +/- 35 cm-1. The temperature dependence of the observed cysteine C beta H and C alpha H protons (a and h) were fit in a separate experiment providing a Delta EL value of 585 +/- 125 cm-1. The differences between the Delta EL values determined by 1H NMR spectroscopy and those determined by EPR or MCD likely arise from coupling between relatively low-frequency vibrational states and the ground and excited electronic states. PMID- 10460175 TI - Crystal structure of the proenzyme domain of plasminogen. AB - We have solved the X-ray crystal structure of the proenzyme form of the catalytic domain of plasminogen, with the nonessential mutations M585Q, V673M, and M788L, to 2.0 A resolution. The structure presents an inactive protease characterized by Asp740 (chymotrypsinogen 194) hydrogen bonded to His586 (chymotrypsinogen 40), preventing proper formation of the oxyanion hole and S1 specificity pocket. In addition, the catalytic triad residues are misplaced relative to the active conformation adopted by serine proteases in the chymotrypsin family. Finally, a unique form of zymogen inactivation is observed, characterized by a "foot-in mouth" mechanism in which Trp761 (chymotrypsinogen 215) is folded into the S1 specificity pocket preventing substrate binding. PMID- 10460174 TI - Binding properties of the stilbene disulfonate sites on human erythrocyte AE1: kinetic, thermodynamic, and solid state deuterium NMR analyses. AB - A novel stilbene disulfonate, 4-trimethylammonium-4'-isothiocyanostilbene-2,2' disulfonic acid (TIDS), has been chemically synthesized, and the interaction of this probe with human erythrocyte anion exchanger (AE1) was characterized. Covalent labeling of intact erythrocytes by [N(+)((14)CH(3))(3)]TIDS revealed that specific modification of AE1 was achieved only after removal of other ligand binding sites by external trypsinization. Following proteolysis, (1.2 +/- 0.4) x 10(6) TIDS binding sites per erythrocyte could be blocked by prior treatment with 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), a highly specific inhibitor of AE1. Inhibition of sulfate equilibrium exchange by TIDS in whole cells was described by a Hill coefficient of 1.10 +/- 0.06, which reduced to 0.51 +/- 0.01 following external trypsinization. The negative cooperativity of TIDS binding following external trypsinization suggests that trypsin-sensitive proteins modulate allosteric coupling between AE1 monomers. Thermodynamic analysis revealed that TIDS binding induces smaller conformational changes in AE1 than is observed following DIDS binding. The similar inhibitory potencies of both TIDS (IC(50) = 0.71 +/- 0.48 microM) and DIDS (IC(50) = 0.2 microM) imply that there is no correlation between the ability of stilbene disulfonates to arrest anion exchange function and the magnitude of ligand-induced conformational changes in AE1. Solid state (2)H NMR analysis of a [N(+)(CD(3))(3)]TIDS-AE1 complex in both unoriented and macroscopically oriented membranes revealed that large amplitude "wobbling" motions describe ligand dynamics. The data are consistent with a model where TIDS bound to AE1 is located exofacially in contact with the bulk aqueous phase. PMID- 10460176 TI - Amyloid beta protein (1-40) forms calcium-permeable, Zn2+-sensitive channel in reconstituted lipid vesicles. AB - Amyloid beta protein (A beta P) forms senile plaques in the cerebrocortical blood vessels and brain parenchyma of patients with Alzheimer's disease (AD). The nonfamilial or sporadic AD (SAD), the most prevalent form of AD, has been correlated with an increased level of 40-residue A beta P (A beta P1-40). However, very little is known about the role of A beta P1-40 in AD pathophysiology. We have examined the activity of A beta P1-40 reconstituted in phospholipid vesicles. A combined light fluorescence and atomic force microscope (AFM) was used to image the structure of reconstituted vesicles and 45Ca2+ uptake was used as an assay for calcium permeability across the vesicular membrane. Vesicles reconstituted with fresh and globular A beta P1-40 contain a significant amount of A0 beta P and exhibit strong immunofluorescence labeling with an antibody raised against the N-terminal domain of A beta P, suggesting the incorporation of A beta P1-40 peptide in the vesicular membrane. Vesicles reconstituted with A beta P1-40 exhibited a significant level of 45Ca2+ uptake. The vesicular calcium level saturated over time, showing an important ion channel characteristic. The 45Ca2+ uptake was inhibited by (i) a monoclonal antibody raised against the N-terminal region of A beta P and (ii) Zn2+. However, a reducing agent (DTT) did not inhibit the 45Ca2+ uptake, indicating that the oxidation of A beta P or its surrounding lipid molecules is not directly involved in A beta P-mediated Ca2+ uptake. These findings provide biochemical and structural evidence that fresh and globular A beta P1-40 forms calcium-permeable channels and thus may induce cellular toxicity by regulating the calcium homeostasis in nonfamilial or sporadic Alzheimer's disease. PMID- 10460177 TI - Thermodynamic, spectroscopic, and equilibrium binding studies of DNA sequence context effects in six 22-base pair deoxyoligonucleotides. AB - Effects of different end sequences on stability, circular dichroism spectra (CD), and enzyme binding properties were investigated for six 22-base pair, non-self complementary duplex DNA oligomers. The center sequences of these deoxyoligonucleotides have 8-14 base pairs in common and are flanked on both sides by sequences differing in context and A-T content. Temperature-induced melting transitions monitored by differential scanning calorimetry (DSC) and ultraviolet absorbance were measured for the six duplexes in buffered 115 mM Na(+) solutions. Values of the melting transition enthalpy, DeltaH(cal), and entropy, DeltaS(cal), were obtained directly from DSC experiments. Melting transition parameters, DeltaH(vH) and DeltaS(vH), were also estimated from van't Hoff analysis of optical melting curves collected as a function of DNA concentration, assuming a two-state melting transition. Melting free energies (20 degrees C) of the six DNAs evaluated from DSC experiments ranged from -18.7 to 32.7 kcal/mol. van't Hoff estimates of the free energies ranged from -18.5 to 48.0 kcal/mol. With either method, the trends in free energy as a function of sequence were identical. Equilibrium binding by BamHI restriction endonuclease to the 22-base pair DNAs was also investigated. The central eight base pairs of all six molecules, 5'-A-GGATCC-A-3', contained a BamHI recognition sequence bounded by A-T base pairs. Magnesium free binding assays were performed by titering BamHI against a constant concentration of each of the deoxyoligonucleotide substrates and analyzing reaction products by gel retardation. Binding isotherms of the total amount of bound DNA versus protein concentration were constructed which provided semiquantitative estimates of the equilibrium dissociation constants for dissociation of BamHI from the six DNA oligomers. Dissociation constants ranged from 0.5 x 10(-)(9) to 12.0 x 10(-)(9) M with corresponding binding free energies of -12.5 to -10.6 (+/-0. 1) kcal/mol. An inverse relationship is found when binding and stability are compared. PMID- 10460178 TI - Purification, cloning, and synthesis of a novel salivary anti-thrombin from the mosquito Anopheles albimanus. AB - An anti-thrombin peptide (anophelin) was isolated from the salivary glands of the mosquito Anopheles albimanus through molecular sieving and reverse-phase high performance liquid chromatography. The purified peptide inhibited thrombin induced platelet aggregation, thrombin esterolytic activity on a synthetic substrate, and thrombin cleavage of fibrinogen. The purified anti-thrombin had a molecular mass of 6342.4 Da. Its amino terminus was blocked, but internal sequence yielded three peptide sequences, which were used to design oligonucleotide probes for polymerase chain reaction amplification of salivary gland cDNA and isolation of the full-length clone. Analysis of the sequence of anophelin shows no similarities to any other anti-thrombin peptides. Anophelin was successfully synthesized and characterized to be a tight-binding, specific, and novel inhibitor of thrombin. PMID- 10460179 TI - The peculiar nature of the guanidine hydrochloride-induced two-state denaturation of staphylococcal nuclease: a calorimetric study. AB - This work determines the ratio of DeltaH(vH) /DeltaH(cal) for staphylococcal nuclease (SN) denaturation in guanidine hydrochloride (GdnHCl) to test whether GdnHCl-induced denaturation is two-state. Heats of mixing of SN as a function of [GdnHCl] were determined at pH 7.0 and 25 degrees C. The resulting plot of DeltaH(mix) vs [GdnHCl] exhibits a sigmoid shaped curve with linear pre- and post denaturational base lines. Extending the pre- and post-denaturational lines to zero [GdnHCl] gives a calorimetric DeltaH (DeltaH(cal)) of 24.1 +/- 1.0 kcal/mol, for SN denaturation in the limit of zero GdnHCl concentration. Guanidine hydrochloride-induced denaturation Gibbs energy changes in the limit of zero denaturant concentration (DeltaG degrees (N)(-)(D)) at pH 7. 0 were determined for SN from fluorescence measurements at fixed temperatures over the range from 15 to 35 degrees C. Analysis of the resulting temperature-dependent DeltaG degrees (N)(-)(D) data defines a van't Hoff denaturation enthalpy change (DeltaH(vH)) of 26. 4 +/- 2.8 kcal/mol. The model-dependent van't Hoff DeltaH(vH) divided by the model-independent DeltaH(cal) gives a ratio of 1.1 +/- 0.1 for DeltaH(vH)/DeltaH(cal), a result that rules out the presence of thermodynamically important intermediate states in the GdnHCl-induced denaturation of SN. The likelihood that GdnHCl-induced SN denaturation involves a special type of two state denaturation, known as a variable two-state process, is discussed in terms of the thermodynamic implications of the process. PMID- 10460180 TI - A focus of deer tick virus transmission in the northcentral United States. AB - We screened salivary glands from adult deer ticks collected near Spooner and Hayward, Wisconsin, to determine whether deer tick virus, a recently described flavivirus, occurs with other tickborne agents in the upper Midwest. Intraacinar inclusions suggestive of replicating virus were detected in 4 (4.6%) of 87 ticks. The virus was isolated by suckling-mouse inoculation. PMID- 10460181 TI - Dengue reemergence in Argentina. AB - Aedes aegypti, eradicated from Argentina in 1963, has now reinfested the country as far south as Buenos Aires. In 1997, four persons with travel histories to Brazil, Ecuador, or Venezuela had confirmed dengue, and surveillance for indigenous transmission allowed the detection of 19 dengue cases in Salta Province. These cases of dengue are the first in Argentina since 1916 and represent a new southern extension of dengue virus. PMID- 10460182 TI - Non-host cells in the pathogenesis of autoimmune disease: a new paradigm? PMID- 10460183 TI - Pulse methylprednisolone therapy for arthritis causing muscle weakness. PMID- 10460184 TI - Does social disadvantage contribute to the excess mortality in rheumatoid arthritis patients? AB - BACKGROUND: Socioeconomic deprivation is associated with increased mortality from cardiovascular causes and malignancy. The influence of disadvantage in patients with rheumatoid arthritis (RA), who are known to have premature mortality, has not been ascertained. AIM: To assess the relation between the level of patient deprivation and mortality in RA patients. METHODS: 200 RA patients, enrolled in a study comparing sulfasalazine and penicillamine in 1984-85, have been followed up prospectively for 12 years. Subjects were categorised into Carstairs groups with deprivation scores ranging from 1 (most affluent) to 7 (most deprived). Information about deaths was obtained from the Registrar General in Scotland, death certificates and GP/hospital records. RESULTS: There were more RA patients in the deprived areas then expected compared with the West of Scotland and England and Wales. Some 47.5% of the RA patients had died by 12 years-the majority of cardiorespiratory causes or malignancy. There were no differences in the median age or disease duration in the various Carstairs groups at study entry, but the percentage of deaths was higher in the more deprived groups after 12 years (36% dead in most affluent area compared with 61% in the most deprived, that is, in groups 6 and 7). CONCLUSION: In patients with RA increasing deprivation was associated with premature mortality. If confirmed elsewhere these findings have implications for rheumatological management strategies, for researchers involved in collaborative studies of patients from different socioeconomic backgrounds and for resource allocation. PMID- 10460185 TI - No effect of bipolar interferential electrotherapy and pulsed ultrasound for soft tissue shoulder disorders: a randomised controlled trial. AB - OBJECTIVE: To assess the efficacy of bipolar interferential electrotherapy (ET) and pulsed ultrasound (US) as adjuvants to exercise therapy for soft tissue shoulder disorders (SD). METHODS: Randomised placebo controlled trial with a two by two factorial design plus an additional control group in 17 primary care physiotherapy practices in the south of the Netherlands. Patients with shoulder pain and/or restricted shoulder mobility, because of a soft tissue impairment without underlying specific or generalised condition, were enrolled if they had not recovered after six sessions of exercise therapy in two weeks. They were randomised to receive (1) active ET plus active US; (2) active ET plus dummy US; (3) dummy ET plus active US; (4) dummy ET plus dummy US; or (5) no adjuvants. Additionally, they received a maximum of 12 sessions of exercise therapy in six weeks. Measurements at baseline, 6 weeks and 3, 6, 9, and 12 months later were blinded for treatment. OUTCOME MEASURES: recovery, functional status, chief complaint, pain, clinical status, and range of motion. RESULTS: After written informed consent 180 patients were randomised: both the active treatments were given to 73 patients, both the dummy treatments to 72 patients, and 35 patients received no adjuvants. Prognosis of groups appeared similar at baseline. Blinding was successfully maintained. At six weeks seven patients (20%) without adjuvants reported very large improvement (including complete recovery), 17 (23%) and 16 (22%) with active and dummy ET, and 19 (26%) and 14 (19%) with active and dummy US. These proportions increased to about 40% at three months, but remained virtually stable thereafter. Up to 12 months follow up the 95% CI for differences between groups for all outcomes include zero. CONCLUSION: Neither ET nor US prove to be effective as adjuvants to exercise therapy for soft tissue SD. PMID- 10460186 TI - Decreased serum biochemical markers of muscle origin in patients with ankylosing spondylitis. AB - OBJECTIVES: There is a lack of unanimity about (increased) serum levels of creatine kinase (CK) in patients with ankylosing spondylitis (AS), perhaps because of the inclusion of inappropriate controls. Therefore, serum levels of biochemical markers of muscle origin were assessed in AS patients compared with controls. METHODS: In a comparative study serum levels of sarcoplasmic proteins indicating muscle cell leakage, creatinine, and C reactive protein (CRP) were measured. Fifty eight AS patients with a mean disease duration of 22 (SD 11) years and 58 age and sex matched controls (without back complaints) were included. RESULTS: Lower serum levels in AS patients compared with controls were found for CK (mean (SD): 46 (21) v 76 (44) IU/l; p<0.001), aldolase (0.43 (0.36) v 0.58 (0. 32) IU/l; p=0.001), creatinine (91 (13) v 96 (11) micromol/l; p=0. 02), alanine aminotransferase (2.8 (1.5) v 4.1 (2.9) IU/l; p=0.001) and aspartate aminotransferase (7.0 (2.7) v 8.4 (3.5) IU/l; p=0.02). Also the lean body mass, as estimated by a formula using height, weight, age and sex, showed lower values in patients versus controls (56 (9) v 59 (9) kg; p=0.004), but creatinine clearance (by Cockcroft and Gault formula) was not different (p=0.48). Partial correlation coefficients adjusted for age and sex showed that CRP levels correlated negatively with CK and aldolase levels in AS patients (r= -0.48, p<0.001 and r= -0.37, p=0.005, respectively). CONCLUSION: Serum levels of biochemical markers of muscle origin were lower in AS patients compared with controls. Patients with active AS, as reflected by high CRP levels, may have an increased protein degradation, predominantly in skeletal muscle. PMID- 10460187 TI - Characterisation of T cell clonotypes that accumulated in multiple joints of patients with rheumatoid arthritis. AB - OBJECTIVE: To investigate whether identical T cell clonotypes accumulate in multiple rheumatoid joints, the clonality of T cells that had infiltrated into synovial tissue (ST) samples simultaneously obtained from multiple joints of patients with rheumatoid arthritis (RA) was analysed. METHODS: T cell receptor (TCR) beta gene transcripts, amplified by reverse transcription-polymerase chain reaction from ST and peripheral blood lymphocytes of five RA patients, were subjected to single strand conformation polymorphism analysis and DNA sequencing. RESULTS: Approximately 40% of accumulated T cell clonotypes found in one joint of a patient were found in multiple joints in the same patient. Furthermore, identical amino acid sequences were found in TCR beta junctional regions of these clonotypes from different patients with at least one HLA molecule match. CONCLUSIONS: The T cell clonotypes accumulating in multiple rheumatoid joints may be involved in the perpetuation of polyarthritis by reacting to antigens common to these multiple joints. PMID- 10460188 TI - Double blind glucocorticoid controlled trial of samarium-153 particulate hydroxyapatite radiation synovectomy for chronic knee synovitis. AB - BACKGROUND: Samarium-153 particulate hydroxyapatite (Sm-153 PHYP) is a relatively new radiation synovectomy agent developed for the treatment of chronic synovitis. Although it has been shown that the levels of unwanted extra-articular radiation are lower after intra-articular injection of Sm-153 PHYP than yttrium-90 colloid, its clinical efficacy has not been rigorously studied. OBJECTIVES: To establish whether Sm-153 PHYP radiation synovectomy results in a clinically useful benefit sustained at one year. METHODS: In a randomised double blind study, patients received either intra-articular 40 mg triamcinolone hexacetonide alone or 40 mg triamcinolone hexacetonide combined with Sm-153 PHYP in an outpatient clinic. RESULTS: Sixty patients (28 male, 32 female), median age 51 (18-75) with chronic knee synovitis were studied. Diagnoses included: rheumatoid arthritis (n=29); psoriatic arthritis (n=9); ankylosing spondylitis (n=3); reactive arthritis (n=2); undifferentiated seronegative oligoarthritis (n=13) and miscellaneous inflammatory conditions (n=4). More patients who received Sm-153 PHYP/triamcinolone hexacetonide sustained clinical benefit a year after treatment compared with patients who received corticosteroid alone (12 of 31 (39%) v 6 of 29 (21%), a difference of 18% more patients (95% CI -5% to 41%)) though the difference was not significant (chi(2)=2.31, 0.2>p>0.1, n=60). Despite the variation in injected activity (median 563 MBq, range 218-840 MBq), there was no obvious relation between low levels of injected activity (<555 MBq) and relapse within 12 months of treatment (chi(2) =2.61, 0.2>p>0.1, n=31). CONCLUSIONS: There was no clear beneficial clinical effect of combined Sm-153 PHYP/triamcinolone hexacetonide injection over triamcinolone hexacetonide alone a year after treatment for chronic knee synovitis. PMID- 10460189 TI - The value of synovial fluid analysis in the assessment of knee joint destruction in arthritis in a three year follow up study. AB - OBJECTIVES: To assess the predictive significance of synovial fluid (SF) analysis for progressive radiological knee joint destruction in arthritis. METHODS: Altogether 55 patients with arthritis and knee joint effusion were included in the study. The diagnosis was rheumatoid arthritis (RA) for 44 of them, chronic seronegative spondylarthropathy for seven and juvenile rheumatoid arthritis for four. The mean age of the patients was 51.8 (SD 14.9, range 19-82) years, and the mean duration of disease 10.9 (SD 9.2, range 0.5-37) years. In addition to the routine laboratory tests, different markers of collagen synthesis and breakdown in serum and SF were assessed. The radiological grade of the knee joint was assessed by Larsen's method at the baseline and after a three year follow up. RESULTS: During the follow up, Larsen's grade deteriorated in 22 (40%) patients. These patients had a significantly higher median level of cross linked carboxyterminal telopeptide of type I collagen (ICTP) in SF at entry than those who had a stable index (p = 0.035). Serum ICTP did not have any predictive value for a specific joint. The median levels of total SF leucocytes (p = 0.012) and the subgroup of polymorphonuclear leucocytes (p = 0.018) were higher in the patients with a stable Larsen's index. However, the relation of SF leucocyte level to radiological progression could not be confirmed in the RA group. CONCLUSION: It is concluded that SF analysis may help in the identification of patients with inflammatory arthritis who are at risk for progressive destruction in a particular joint. A high total SF leucocyte level is not necessarily associated with a poor prognosis. Instead, a high SF ICTP level seems to reflect accelerated bone degradation. PMID- 10460190 TI - Rasch analysis of the Western Ontario MacMaster questionnaire (WOMAC) in 2205 patients with osteoarthritis, rheumatoid arthritis, and fibromyalgia. AB - OBJECTIVE: Advances in health measurement have led to the application of Rasch Item Response Theory (IRT) analysis (Rasch analysis) to evaluate instruments measuring health status and quality of life of patients, including the Health Assessment Questionnaire and SF-36. This study investigated the extent to which the Western Ontario MacMaster osteoarthritis questionnaire (WOMAC) satisfies the Rasch model, particularly in respect to unidimensionality, item separation, and linearity. METHODS: The study included a total of 2205 patients, 1013 with rheumatoid arthritis (RA), 655 with osteoarthritis of the knee or hip (OA), and 537 with fibromyalgia. All patients completed the WOMAC as part of a longitudinal study of rheumatic disease outcomes. To examine whether the WOMAC pain and function scales each fits the Rasch model, the Winsteps program was used to assess item difficulty, scale unidimensionality, item separation, and linearity. RESULTS: Although the WOMAC worked best in OA, regardless of disorder, both the pain and function scales were unidimensional, had adequate item separation, and had a long range (25-150) of linearity in the function scale. Several functional items, however, had a high information weight fit (INFIT) statistic, indicating poor fit to the model. These items included "getting in and out of the bath" and "going down stairs." CONCLUSION: The WOMAC generally satisfies the requirements of Rasch item response theory across all disorders studied, and is an appropriate measure of lower body function in OA, RA and fibromyalgia. Although some individual items do not fit well, it is not likely that removing such items would result in more than overall minimal differences, and it will be difficult to remove traces of multidimensionality while keeping the central constructs of progressive lower body musculoskeletal abnormality intact. In addition, it is possible that a "purer", still more unidimensional instrument would be less useful in clinical trials and epidemiological studies by restricting the range of the scale. PMID- 10460191 TI - Projecting the need for hip replacement over the next three decades: influence of changing demography and threshold for surgery. AB - OBJECTIVES: To estimate the requirement for total hip replacement in the United Kingdom over the next three decades METHODS: Projection of age and sex specific hip replacements in the UK over 10 year intervals taking account of demographic change and the extrapolation of arthroplasty rates from Sweden; a country with recently introduced guidelines. RESULTS: Assuming no change in the age and sex specific arthroplasty rates, the estimated number of hip replacements will increase by 40% over the next 30 year period because of demographic change alone. The proportionate change will be substantially higher in men (51%) than women (33%), with a doubling of the number of male hip replacements in those aged over 85. Changes in the threshold for surgery may increase this further-up to double the current number. CONCLUSION: A sharp rise in hip replacements will be needed to satisfy needs in the UK population over the next 30 years. PMID- 10460193 TI - Most calcium pyrophosphate crystals appear as non-birefringent. AB - OBJECTIVE: To determine the proportion of calcium pyrophosphate dihydrate (CPPD) crystals that appear as non-birefringent when observed under the polarised light microscope. METHODS: Two observers examined independently 10 synovial fluid samples obtained during an episode of arthritis attributable to CPPD crystals. Ten synovial fluid samples from patients with acute gout were used as a reference. The examination was performed after placing a fluid sample in a Niebauer haemocytometric chamber; a crystal count was done first under ordinary light, then in the area corresponding to a 0.1 ml, under polarised light RESULTS: The percentages of birefringence appreciated for CPPD were 18% (confidence intervals (CI) 12, 24) for observer 1, and 17% (CI 10, 24) for observer 2 (difference NS). The percentages of birefringence for monosodium urate were 127% (CI 103, 151) for observer 1 and 107% (CI 100, 114) for observer 2 (difference NS). Percentages above 100% indicate that crystals missed under ordinary light became apparent under polarised light. CONCLUSION: Only about one fifth of all CPPD crystals identified by bright field microscopy show birefringence when the same synovial fluid sample is observed under polarised light. If a search for CPPD crystals is conducted under polarised light, the majority of the crystals will be missed. Ordinary light allows a better rate of CPPD crystal detection but observation under polarised light of crystals showing birefringence is required for definitive CPPD crystal identification. PMID- 10460192 TI - Tumour necrosis factor alpha stimulated rheumatoid synovial microvascular endothelial cells exhibit increased shear rate dependent leucocyte adhesion in vitro. AB - OBJECTIVE: To investigate endothelial cell adhesion molecule expression and leucocyte adhesion to endothelial cells isolated from the microvasculature of rheumatoid arthritic synovial tissue (SMEC) in comparison with similar cells isolated from healthy subcutaneous adipose tissue (ADMEC) or from umbilical veins (HUVEC). METHODS: Cultured endothelial cells were treated with tumour necrosis factor alpha (TNFalpha) for 2-24 hours before the assessment of cell surface E selectin, vascular (VCAM-1) or intercellular cell adhesion molecule-I (ICAM-1) expression. Neutrophil and T lymphocyte adhesion to TNFalpha treated endothelial cells was assessed using static and shear dependent assay systems. RESULTS: VCAM 1 expression by SMEC was significantly less sensitive to TNFalpha stimulation than HUVEC or ADMEC. E-selectin expression by SMEC appeared to be more sensitive to TNFalpha stimulation and maximal expression was about 30% greater in comparison with HUVEC or ADMEC. Sensitivity to TNFalpha induction and maximal ICAM-1 expression was similar in all three endothelial cell types. Static neutrophil adhesion to TNFalpha stimulated SMEC was significantly increased in comparison with HUVEC, however this phenomenon was dependent on the presence of neutralising antibodies to ICAM-1. At shear rates in excess of 2.4 dynes/cm(2) significantly more neutrophils and, predominantly CD45RO+, T lymphocytes adhered to TNFalpha stimulated SMEC than HUVEC. CONCLUSION: Rheumatoid synovial endothelial cells differentially regulate E-selectin and VCAM-1. The increased ability of TNFalpha stimulated synovial endothelial cells to support leucocyte adhesion may help to explain the leucocyte, in particular CD45RO+ T-lymphocyte, recruitment observed in the rheumatoid synovium. PMID- 10460194 TI - Old drug, new tricks: haloperidol inhibits secretion of proinflammatory cytokines. AB - OBJECTIVES: It was noted that treatment of a patient with acute mania by haloperidol was associated with marked improvement in activity of rheumatoid arthritis. The objective of this study was to examine the effects of haloperidol on inflammatory cytokine release in vitro, as a potential mechanism to explain the in vivo anti-inflammatory effects of haloperidol. METHODS: The effect of haloperidol on the production of inflammatory cytokines interleukin 1beta (IL1beta) and tumour necrosis factor alpha (TNFalpha) was measured in bacterial lipopolysaccharide stimulated whole blood cultures and on the promonocyte cell line THP-1, using commercial and in house enzyme linked immunosorbent assays to measure cytokine concentrations. RESULTS: Haloperidol inhibited lipopolysaccharide stimulated production of both IL1beta and TNFalpha in vitro in a dose dependent manner and over a prolonged time period. Marked inhibition was seen over a range of concentrations of haloperidol from 0.5 microgram/ml to 50 microgram/ml, including those predicted to occur in the patient's blood. CONCLUSIONS: Haloperidol treatment seemed to alleviate inflammation in rheumatoid arthritis. In vitro experiments would suggest that the mechanism is by direct inhibition of proinflammatory cytokine release. This phenomenon requires further investigation and may potentially lead to the development of novel treatment. PMID- 10460195 TI - Vitamin A-sensitive tissues in transgenic mice expressing high levels of human cellular retinol-binding protein type I are not altered phenotypically. AB - The suggested function of cellular retinol-binding protein type I [CRBP(I)] is to carry retinol to esterifying or oxidizing enzymes. The retinyl esters are used in storage or transport, whereas oxidized forms such as all-trans or 9-cis retinoic acid are metabolites used in the mechanism of action of vitamin A. Thus, high expression of human CRBP(I) [hCRBP(I)] in transgenic mice might be expected to increase the production of retinoic acid in tissues, thereby inducing a phenotype resembling vitamin A toxicity. Alternatively, a vitamin A-deficient phenotype could also be envisioned as a result of an increased accumulation of vitamin A in storage cells induced by a high hCRBP(I) level. Signs of vitamin A toxicity or deficiency were therefore examined in tissues from transgenic mice with ectopic expression of hCRBP(I). Testis and intestine, the tissues with the highest expression of the transgene, showed normal gross morphology. Similarly, no abnormalities were observed in other tissues known to be sensitive to vitamin A status such as cornea and retina, and the epithelia in the cervix, trachea and skin. Furthermore, hematologic variables known to be influenced by vitamin A status such as the hemoglobin concentration, hematocrits and the number of red blood cells were within normal ranges in the transgenic mice. In conclusion, these transgenic mice have normal function of vitamin A despite high expression of hCRBP(I) in several tissues. PMID- 10460196 TI - Soybean phytochemicals inhibit the growth of transplantable human prostate carcinoma and tumor angiogenesis in mice. AB - The objectives of our studies are to characterize the ability of dietary soybean components to inhibit the growth of prostate cancer in mice and alter tumor biomarkers associated with angiogenesis. Soy isoflavones (genistein or daidzein) or soy phytochemical concentrate inhibit the growth of prostate cancer cells LNCaP, DU 145 and PC-3 in vitro, but only at supraphysiologic concentrations, i.e., 50% inhibitory concentration (IC(50)) > 50 micromol/L. G2-M arrest and DNA fragmentation consistent with apoptosis of prostate cancer cells are also observed at concentrations causing growth inhibition. In contrast, the in vitro proliferation of vascular endothelial cells was inhibited by soy phytochemcials at much lower concentrations. We evaluated the ability of dietary soy phytochemical concentrate and soy protein isolate to inhibit the growth of the LNCaP human prostate cancer in severe combined immune-deficient mice. Mice inoculated subcutaneously with LNCaP cells (2 x 10(6)) were randomly assigned to one of the six dietary groups based on the AIN-76A formulation for 3 wk. A 2 x 3 factorial design was employed with two protein sources (20%, casein vs. soy protein) and three levels of soy phytochemical concentrate (0, 0.2 and 1.0% of the diet). Soy components did not alter body weight gain or food intake. Compared with casein-fed controls, the tumor volumes after 3 wk were reduced by 11% (P = 0.45) by soy protein, 19% (P = 0.17) by 0.2% soy phytochemical concentrate, 28% by soy protein with 0.2% soy phytochemical concentrate (P < 0.05), 30% by 1.0% soy phytochemical concentrate (P < 0.05) and 40% by soy protein with 1.0% soy phytochemical concentrate (P < 0.005). Histologic examination of tumor tissue showed that consumption of soy products significantly reduced tumor cell proliferation, increased apoptosis and reduced microvessel density. The angiogenic protein insulin-like growth factor-I was reduced in the circulation of mice fed soy protein and phytochemical concentrate. Our data suggest that dietary soy products may inhibit experimental prostate tumor growth through a combination of direct effects on tumor cells and indirect effects on tumor neovasculature. PMID- 10460197 TI - Regular exercise modulates muscle membrane phospholipid profile in rats. AB - We investigated the effect of regular exercise and changes in dietary fatty acid profile on skeletal muscle phospholipid fatty acid profile in rats. Rats were randomly divided into three groups and for 4 wk fed either a carbohydrate-rich diet (CHO, 10 percent of total energy (E%) fat, 20 E% protein, 70 E% CHO) or one of two fat-rich diets (65 E% fat, 20 E% protein, 15 E% CHO) containing predominantly either saturated or monounsaturated fatty acids. Each dietary group was randomly assigned to a trained (6 d/wk, progressive to 60 min, 28 m/min at a 10 degrees incline) or a sedentary group. The effect of training was apparent in the three hindlimb muscles analyzed: red quadriceps, white quadriceps and soleus. The unsaturation index was significantly lower in the trained than in the sedentary groups (206 +/- 2 vs. 215 +/- 2, P < 0. 01), which largely reflected a lower content of arachidonic acid [20:4(n-6): 14.5 +/- 0.5 vs. 16.6 +/- 0.4% of total fatty acids, P < 0.01] and docosahexaenoic acid [22:6(n-3): 11.1 +/- 0.2 vs. 11.7 +/- 0.3% of total fatty acids, P < 0.03] and a concomitant higher content of linoleic acid [18:2(n-6): 20.0 +/- 0.4 vs. 17.8 +/- 0.4% of total fatty acids, P < 0.01]. Training affected skeletal muscle membrane structural composition, and this occurred independently of dietary fatty acid changes. This change likely reflects an increased utilization of highly unsaturated fatty acids for energy, an effect which may have deleterious effects on insulin action. PMID- 10460198 TI - Expression of the mouse metallothionein-I and -II genes provides a reproductive advantage during maternal dietary zinc deficiency. AB - The function of metallothionein in zinc homeostasis was examined by using mice homozygous for knockout (KO) of the metallothionein-I or -II (MT-I and MT-II) genes. Pregnant MT-I/II KO mice or control mice were fed a zinc-deficient (1 microg/g or 5 microg/g) diet or a zinc-adequate (50 microg/g) diet during specific periods of pregnancy, and the effects on morphogenesis of the embryos were determined at day 14 of pregnancy (day 1 = vaginal plug). In the homozygous MT-I/II KO, as well as in the nontransgenic control mice, severe dietary zinc deficiency (1 microg/g) beginning on day 1 of pregnancy was embryotoxic and teratogenic, and the majority of the embryos in both strains were dead by mid gestation. However, 53% of the surviving embryos in the MT-I/II KO mice were morphologically abnormal compared to only 32% of the embryos in the control mice. In subsequent experiments, moderate dietary zinc deficiency (5 microg/g beginning on day 1 of pregnancy or 1 microg/g dietary zinc beginning on day 8 of pregnancy) exerted teratogenic, but not embryotoxic effects. Embryos in the MT-I/II KO mice were 260 to 290% as likely to develop abnormally than were embryos in the control mice fed these same diets. These results demonstrate that the expression of the MT-I and -II genes in pregnant females improves reproductive success during maternal dietary zinc deficiency. PMID- 10460199 TI - Delayed changes in postprandial lipid in young normolipidemic men after a nocturnal vitamin A oral fat load test. AB - The oral fat load tests (OFLT) used to study postprandial lipemia are generally conducted during the day. A nocturnal fat load test could be convenient and physiologically more appropriate. We have therefore compared the lipemic responses of 9 normolipidemic young men to OFLT given at 2200 h (nocturnal) and at 0700 h (diurnal). Triglyceride and retinyl palmitate concentrations were measured for 10 h. Peak plasma concentrations or areas under curves (AUC) for triglyceride after the diurnal and nocturnal tests were not significantly different [2.17 +/- 0.78 (diurnal) vs. 2.04 +/- 0.87 mmol/L (nocturnal) and 13.12 +/- 4.45 (diurnal) vs. 13.74 +/- 5.79 mmol/(L. h) (nocturnal)]. Peak plasma concentrations and AUC retinyl palmitate for the two tests were not different [1.71 +/- 0.69 (diurnal) vs. 1.42 +/- 0.66 mg/L (nocturnal) and 7.17 +/- 3.98 (diurnal) vs. 6.63 +/- 4.23 mg/(L. h) (nocturnal)]. The diurnal triglyceride peak occurred significantly earlier (4.3 +/- 1.2 h) than the nocturnal peak (5.8 +/- 1.7 h, P < 0.05). We have developed a model using only three sample time points to predict AUC [triglyceride at 0 h, triglyceride at average peak-time (4 h for diurnal and 6 h for nocturnal tests), and triglyceride at 10 h], thus reducing the number of blood samples. The predicted AUC was well correlated with the total AUC after nocturnal OFLT (r = 0.98, P < 0.0001). The nocturnal test appeared to be well tolerated by the subjects. The three-point simplified protocol may well be suitable for studies on large groups of subjects. PMID- 10460200 TI - A common mutation A1298C in human methylenetetrahydrofolate reductase gene: association with plasma total homocysteine and folate concentrations. AB - Methylenetetrahydrofolate reductase (MTHFR) is one of the main regulatory enzymes of homocysteine metabolism. Previous studies revealed that a common mutation in MTHFR gene C677T is related to hyperhomocysteinemia and occlusive vascular pathology. In the current study, we determined the prevalence of a newly described mutation in the human MTHFR gene A1298C, and the already known C677T mutation, and related them to plasma total homocysteine and folate concentrations. We studied 377 Jewish subjects, including 190 men and 186 women aged 56.8 +/- 13 y (range 32-95 y). The frequency of the homozygotes for the A1298C and the C677T MTHFR mutations was common in the Jewish Israeli population (0.34 and 0.37, respectively). Subjects homozygous (TT) for the C677T mutation had significantly greater plasma total homocysteine concentrations (P < 0.01) than subjects without the mutation (CC). Homozygotes (CC) for the A1298C mutation did not have elevated plasma total homocysteine concentrations. Our study indicated that subjects with the 677CC/1298CC genotype had significantly lower concentrations (P < 0. 05) than those with a 677CC/1298AA genotype. Neither mutation (the A1298C and the C677T) was associated with established cardiovascular risk factors such as hypertension, elevated total cholesterol or body mass index. PMID- 10460201 TI - Catechin is present as metabolites in human plasma after consumption of red wine. AB - Flavonoids are components of fruits, vegetables and wines. An abundance of flavonoids in the diet is correlated with reduced heart disease mortality, suggesting that they act as protective nutrients. However, little is known about the absorption and metabolism of flavonoids after normal foods are consumed. This study measured the levels of one abundant flavonoid, (+)-catechin, and its metabolites in plasma after five male and four female volunteers consumed 120 mL of red wine (RW) one day and de-alcoholized red wine (DRW) on a separate day. Each wine sample contained 35 +/- 1 mg catechin (mean +/- SEM). Plasma levels of catechin and its metabolite 3'-O-methylcatechin (3'MC) were determined by gas chromatography-mass spectrometry (GC-MS) of the trimethylsilylated (TMS) derivatives. Glucuronide and sulfate conjugates were determined after enzymatic hydrolysis. Before RW or DRW consumption, plasma levels of catechin, 3'MC and all conjugates were <2 nmol/L. After 1 h, average levels of catechin, 3'MC and all conjugates increased to 91 +/- 14 nmol/L (RW) and 81 +/- 11 nmol/L (DRW). At 1 h, 21 +/- 1% of the metabolites were methylated and <2% of catechin and 3'MC were unconjugated. Catechin was present as both a sulfate conjugate and a conjugate containing both glucuronide and sulfate residues. 3'MC was present primarily as a glucuronide conjugate. At every time point, catechin was present almost exclusively as metabolites, and these levels were independent of ethanol. Therefore, if flavonoids are protective nutrients, the active forms are likely to be metabolites, which are far more abundant in plasma than the forms that exist in foods. PMID- 10460202 TI - World Health Organization hemoglobin cut-off points for the detection of anemia are valid for an Indonesian population. AB - The study was designed to determine whether population-specific hemoglobin cut off values for detection of iron deficiency are needed for Indonesia by comparing the hemoglobin distribution of healthy young Indonesians with that of an American population. This was a cross-sectional study in 203 males and 170 females recruited through a convenience sampling procedure. Hemoglobin, iron biochemistry tests and key infection indicators that can influence iron metabolism were analyzed. The hemoglobin distributions, based on individuals without evidence of clear iron deficiency and infectious process, were compared with the National Health and Nutrition Survey (NHANES) II population of the United States. Twenty percent of the Indonesian females had iron deficiency, but no male subjects were iron deficient. The mean hemoglobin of Indonesian males was similar to the American reference population at 152 g/L with comparable hemoglobin distribution. The mean hemoglobin of the Indonesian females was 2 g/L lower than that of the American reference population, which may be the result of incomplete exclusion of subjects with milder form of iron deficiency. When the WHO cutoff (Hb < 120 g/L) was applied to female subjects, the sensitivity of 34.2% and specificity of 89.4% were more comparable to the test performance for white American women, in contrast to those of the lower cut-off. On the basis of the finding of hemoglobin distribution of men and the test performance of anemia (Hb < 120 g/L) for detecting iron deficiency for women, it is concluded that there is no need to develop different cut-off points for anemia as a tool for iron-deficiency screening in this population. PMID- 10460203 TI - Clinical pallor is useful to detect severe anemia in populations where anemia is prevalent and severe. AB - Clinical pallor is recommended as a simple way to detect severe anemia, but more data are needed on its accuracy and usefulness when assessed by nonphysicians in diverse settings. We measured hemoglobin and trained non-physician health workers to assess clinical pallor of the conjunctiva, palm and nail beds in five population samples in Nepal and Zanzibar, where severe anemia is common. In total, 5,760 individuals were examined, 3,072 of whom were anemic and 192 of whom had severe anemia (hemoglobin <70 g/L). The prevalence of pallor did not correspond to the prevalence of anemia or severe anemia in the groups studied. However, in all studies, pallor at each anatomical site was associated with a significantly lower hemoglobin concentration. The relative performance of different anatomical sites was not consistent among studies, and we recommend that multiple sites be assessed. Pallor at any of the three sites detected severe anemia with >84% specificity. However, the sensitivity varied from 81% in Nepalese postpartum women to 29% in Zanzibari preschoolers in 1996. Overall estimates for sensitivity and specificity were 50 and 92%, respectively. Although imperfect, use of pallor to screen and treat severe anemia by primary care providers is feasible and worthwhile where severe anemia is common. Usually, the majority of persons with severe anemia will be detected at practically no cost. Many people who are not severely anemic will also receive treatment, but the costs of this error are low compared to the benefits. PMID- 10460204 TI - Food restriction beneficially affects renal transport and cortical membrane lipid content in rats. AB - Food restriction (FR) exerts a variety of beneficial effects and may prolong life in both humans and animals. However, studies of its effects on the cortical brush border membrane (BBM) and basolateral membrane (BLM) lipid concentration, which may be pertinent to renal function, have not been reported in detail. We hypothesized that FR would decrease renal work and lower renal membrane lipid concentration. The changes in lipid concentration would be most dramatic in BBM because this membrane is the entry site for the recovery of filtered ions and nutrients. Young male Fischer 344 x Brown-Norway F1 rats consumed food ad libitum (AL) or were food-restricted (FR, 60% of AL consumption) for 6 wk. AL rats had higher fractional excretions of Na(+), K(+), and Cl(-) than did the FR group (P < 0.001). Renal Na,K-ATPase activity in AL rats was 100% higher than in FR rats (P < 0.001), reflecting greater renal work. The work required for renal proton secretion was lower in FR than in the AL rats. In FR rats, all BBM phospholipid concentrations (phosphatidylserine, phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin) were approximately 50% lower than in the AL rats (P < 0.001). In the BLM, food restriction resulted only in lower phosphatidylcholine concentration, while the other phospholipids were unaffected. Plasma and renal membrane (BBM and BLM) cholesterol concentrations were significantly lower in FR than in AL rats. These results show that a nutritionally complete, but energy restricted, diet improves renal function. It also prevents renal membrane lipid deposition and decreases plasma cholesterol. Prolonged food restriction might attenuate the renal injury that occurs in obese humans as a consequence of insulin resistance and atherosclerosis. PMID- 10460205 TI - Carnitine import to isolated hepatocytes and synthesis are accelerated in pivalate-treated rats. AB - To investigate the effect of pivalate on carnitine import and carnitine synthesis in the liver, we measured carnitine uptake in isolated rat hepatocytes with L [(14)C] carnitine and concentrations of free carnitine, gamma-butyrobetaine and acylcarnitines using tandem mass spectrometry. Hepatocytes from rats treated with 20 mmol/L of pivalate for 4 wk had greater L-[(14)C] carnitine uptake than those of unsupplemented rats after 5, 10, 30 and 90 min. Addition of 1 mmol/L of pivalate or 1 mmol/L of pivaloylcarnitine to control cell suspensions did not affect L-[(14)C] carnitine uptake. The K(m) values for L-[(14)C] carnitine uptake for pivalate-treated rats were significantly lower than control (2.9 +/- 0.7 mmol/L for pivalate-treated rats, 6.2 +/- 1.1 mmol/L for controls). The concentration of free carnitine was not reduced in the liver of pivalate-treated rats, whereas the concentrations of acetylcarnitine and gamma-butyrobetaine were significantly lower than controls. In the heart and muscle the concentration of free carnitine was significantly lower and that of gamma-butyrobetaine was higher than controls. These results suggest that carnitine transport from plasma into the liver and synthesis in the liver are accelerated in rats with secondary carnitine deficiency induced by the administration of pivalate. PMID- 10460206 TI - Liquid concentrates are lower in bioavailable tryptophan than powdered infant formulas, and tryptophan supplementation of formulas increases brain tryptophan and serotonin in rats. AB - The bioavailability of tryptophan in powdered and/or liquid concentrate forms of milk-based infant formulas was determined by studying rat growth response by using a slope ratio method (food conversion efficiency: weight gain/food consumed vs. tryptophan consumed). A gelatin basal diet formulated to be adequate in all nutrients, except tryptophan (0.03%), for rat growth was supplemented with graded levels of crystalline L-tryptophan (0.02, 0. 04, 0.06, 0.08, 0.10, 0.12 and 0.14%, standard diets) or infant formulas providing 0.04 and 0.08% supplemental tryptophan (test diets). These diets were fed to weanling rats for 2 wk. Tryptophan bioavailabilities of various formulas varied from 83 to 95%, with some of the liquid concentrates having the lowest values. The levels of bioavailable tryptophan in the liquid concentrate forms (9.7-12.6 mg/g protein) and the powdered forms (11.1-13.1 mg/g protein) were considerably lower than those of human milk (17-19 mg/g protein). Supplementation of the liquid concentrates with graded levels of L-tryptophan (0.1, 0.5 and 1.0%) had no effect on protein quality indices, based on rat growth, but resulted in a dose-related increase in the concentrations of tryptophan in the plasma and brain and of serotonin and 5 hydroxyindole-3-acetic acid in the brains of rats. This study supports further research to investigate the influence of tryptophan supplementation of infant formulas, to more closely simulate tryptophan composition of human milk, on tryptophan metabolites and their potential related effects on sleep latency and neurobehavioral developments in infants. PMID- 10460207 TI - Growing kittens require less dietary calcium than current allowances. AB - We previously demonstrated that a purified diet containing 3.125 microg of cholecalciferol/kg was adequate to maintain plasma concentrations of 25 hydroxyvitamin D in growing kittens. With the use of this concentration of cholecalciferol, the response of growing kittens to varying levels of calcium in purified diets was measured. Five groups (treatments 1-5), each comprised of seven weaned kittens, were given diets containing 3.8, 5.0, 6.0, 7.2 or 8. 1 g calcium/kg diet (Ca:P ratio of 1:1.25) from 9 to 18 wk of age. Two further groups of kittens (treatments 6 and 7) received similar diets containing 6.0 g Ca/kg diet, with Ca:P ratios of 1:1.55 and 1:2.61, respectively. No clinical signs of calcium deficiency were observed, i.e., growth rate, energy intake and plasma total calcium were not affected by the treatments. However, ionized calcium was significantly lower in kittens in treatment 7. Plasma phosphorus was lower in kittens in treatment 7 than in kittens in treatments 1, 2, 3 and 4, and there was a negative relationship between dietary and plasma phosphorus concentrations. Kittens in treatment 7 had a significantly higher alkaline phosphatase concentration in plasma than kittens in treatments 1, 2, 3 and 5. Kittens in treatment 1 had a lower percentage of bone minerals measured by dual-energy X-ray absorptiometry than kittens in treatments 2-6. These results indicate that the calcium requirement of growing kittens is not >6.0 g/kg diet, (calculated metabolizable energy approximately 20 kJ/g) and that kittens are not very sensitive to inverse Ca:P ratios up to 1:1.55. PMID- 10460208 TI - Dietary taurine enhances cholesterol degradation and reduces serum and liver cholesterol concentrations in rats fed a high-cholesterol diet. AB - The effect of taurine on hypercholesterolemia induced by feeding a high cholesterol (HC) diet (10g/kg) to rats was examined. When various amounts of taurine (0.25, 0.5, 1, 2.5, 5, 10, 20, 30, 40 or 50 g/kg diet) were supplemented to HC for 2 wk, serum total cholesterol gradually and significantly decreased in a dose-dependent manner and normalized at the dose of 10 g taurine/kg, compared with the control (cholesterol free) diet group. By contrast, serum HDL cholesterol was elevated by taurine supplementation. The HC diet caused a significant decrease in the concentration of taurine in serum, liver and heart compared to that in the control group, and the effective dose of supplemental taurine to improve its reduction was 2.5 g/kg diet. In the hypercholesterolemic rats fed the HC diet, the excretion of fecal bile acids and hepatic cholesterol 7 alpha-hydroxylase (CYP7A1) activity and its mRNA level increased significantly, and the supplementation of taurine further enhanced these indexes, indicating an increase in cholesterol degradation. The abundance of mRNA for Apo A-I, one of the main components of HDL, was reduced by HC and recovered by taurine supplementation. Agarose gel electrophoresis revealed that, in hypercholesterolemic rats fed the HC diet, the serum level of the heavier VLDL increased significantly, but taurine repressed this increase and normalized this pattern. Significant correlations were observed between the time- and dose dependent increases of CYP7A1 gene expression and the decrease of blood cholesterol concentration in rats fed the HC diet supplemented with taurine (time, r = -0.538, P < 0.01, n = 32; dose, r = -0.738, P < 0.001, n = 20). These results suggest that the hypocholesterolemic effects of taurine observed in the hypocholesterolemic rats fed the HC diet were mainly due to the enhancement of cholesterol degradation and the excretion of bile acid. PMID- 10460209 TI - Changes in women's plasma lipid and lipoprotein concentrations due to moderate consumption of alcohol are affected by dietary fat level. AB - We studied the impact of substituting ethanol for dietary carbohydrate, in high- and low-fat diets, on plasma lipids and lipoprotein concentrations. During a 12 wk, weight maintaining, controlled feeding study, women consumed only food and beverage provided by the Human Studies Facility of the USDA Beltsville Human Nutrition Research Center. Twenty-six women (age 41-59 y) consumed either a high fat diet (38% of energy from fat) or a low-fat diet (18% of energy from fat) for 12 wk. The 12-wk feeding period was divided into two 6-wk periods in a cross-over design during which either ethanol or carbohydrate was added to the diet (5% of total daily energy intake). When the women consuming the high-fat diet had ethanol added to their diet, they had 6% lower plasma cholesterol (P = 0.003), 11% lower LDL cholesterol (P = 0.001) and 3% higher HDL cholesterol (P = 0.06) than when they had an equal amount (% energy) of carbohydrate added to their diet. The greater HDL cholesterol concentration was due to a 21% greater the HDL(2) subfraction (P = 0. 001). The ratio of LDL to HDL cholesterol was 14% lower. No significant differences existed in plasma lipids in women consuming the low-fat diet between the periods in which they had ethanol or carbohydrate added to their diet. This study suggests that the decreases in cardiovascular disease risk factors typically seen with moderate alcohol consumption may not be evident in individuals consuming a diet low in fat. Therefore changes in the risk factors associated with a low-fat diet and moderate alcohol consumption do not appear to be additive. PMID- 10460210 TI - Metabolism of cholesterol is altered in the liver of C3H mice fed fats enriched with different C-18 fatty acids. AB - We examined whether the degree of saturation of C-18 fatty acids influenced hepatic cholesterol metabolism in C3H mice. The mice were fed diets containing 20 g/100 g fat, enriched in stearic (18:0), oleic (18:1) or linoleic acid (18:2) with or without 1 g/100 g cholesterol. Plasma total cholesterol concentration was lower in mice fed the 18:0 diet relative to those fed the 18:1- or 18:2-enriched diets (P < 0.05) regardless of dietary cholesterol supplementation. Dietary cholesterol significantly raised hepatic total cholesterol concentration (P < 0.05) in those fed the 18:1- and 18:2-enriched diets, but not in mice fed the 18:0-enriched diet. Dietary cholesterol raised biliary cholesterol concentration (P < 0. 05) in mice fed the 18:1- and 18:2-enriched diets, but not in mice fed the 18:0-enriched diet. The cholesterol saturation index was variably affected by the fat diets. Feeding diets containing cholesterol suppressed the hepatic 3 hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) activity and induced acyl coenzyme A:cholesterol acyl transferase (ACAT) activity compared with feeding diets without cholesterol (P < 0.05), indicating that the liver was exposed to dietary cholesterol. Hepatic ACAT activity was lower in mice fed the 18:0 enriched diet compared with those fed the 18:1- or 18:2-enriched diets (P < 0.05). Addition of cholesterol to the 18:1 diet induced the largest increase of hepatic ACAT activity, and this was associated with the enrichment of VLDL with cholesterol. Varying the degree of saturation of C-18 fatty acids influences the metabolism and disposition of hepatic cholesterol. PMID- 10460211 TI - Soy protein peptic hydrolysate with bound phospholipids decreases micellar solubility and cholesterol absorption in rats and caco-2 cells. AB - This experiment was designed to evaluate the effects of casein, soy protein, soy protein with bound phospholipids (SP), soy protein peptic hydrolysate (SPH) or soy protein peptic hydrolysate with bound phospholipids (SPHP) on the micellar solubility of cholesterol and the taurocholate binding capacity in vitro. We also evaluated the effects of various proteins on cholesterol metabolism in rats and Caco-2 cells. SPHP had a significantly greater bile acid-binding capacity than that of SPH in vitro. Micellar cholesterol solubility in vitro was significantly lower in the presence of SPHP compared to casein tryptic hydrolysate (CTH). The cholesterol micelles containing SPHP and SPH significantly suppressed cholesterol uptake by Caco-2 cells compared to the cholesterol micelles containing CTH. Consistent with these findings in the in vivo cholesterol absorption study using radioisotopes, fecal excretion of total steroids was significantly greater in rats fed the SPHP diet compared with those fed the casein, soy protein, SP and SPH diets. Serum total cholesterol was significantly lower in rats fed SPHP than in those fed casein. The concentrations of total lipids and cholesterol in liver were significantly lower in the SPHP-fed group compared with all other groups. These results suggest that the suppression of cholesterol absorption by direct interaction between cholesterol-mixed micelles and SPHP in the jejunal epithelia is part of the mechanism underlying the hypocholesterolemic action of SPHP. SPHP may also inhibit the reabsorption of bile acids in the ileum, thus lowering the serum cholesterol level. PMID- 10460212 TI - Chlorella accelerates dioxin excretion in rats. AB - We investigated the effects of Chlorella on fecal excretion of polychlorinated dibenzo-p-dioxin (PCDD) congeners and polychlorinated dibenzofuran (PCDF) congeners in Wistar rats administered the rice oil that caused Yusho disease, as a substitute for purified dioxin. The rats were fed 4 g of a control diet or a 10% Chlorella diet containing 0.2 mL of the rice oil once during the 5-d experimental period. The amounts of PCDD and PCDF congeners excreted in feces from d 1 to 5 in the group fed 10% Chlorella were 0.2-11.3 and 0.3-12.8 times greater (P < 0.05), respectively, than those of the control group. We then investigated the fecal excretion of PCDD and PCDF congeners from d 8 to 35 in rats administered 0.5 mL of the rice oil. Rats consumed the basal diet for 1 wk. After 1 wk, they consumed either the basal diet or the 10% Chorella diet. The fecal excretions of PCDD and PCDF congeners in the group fed 10% Chlorella were 0.3-3.4 and 0.5-2.5 times greater (most, P < 0.05), respectively, than those of the control group. Thus, the fecal excretions of PCDD and PCDF congeners were greater in rats fed Chlorella. These findings suggest that the administration of Chlorella may be useful in preventing gastrointestinal absorption and for promoting the excretion of dioxin already absorbed into tissues. Moreover, these findings suggest that Chlorella might be useful in the treatment of humans exposed to dioxin. PMID- 10460213 TI - Galactose consumption induces conditioned flavor avoidance in rats. AB - Recent findings revealed that intragastric infusions of galactose conditioned a flavor avoidance in adult rats. To determine whether the galactose-conditioned avoidance was due to the infusion procedure, we investigated the flavor conditioning effect of orally consumed galactose. Food-restricted rats drank a flavored galactose solution, a flavored fructose solution and a flavored saccharin solution in separate one-bottle training sessions; grape, cherry and orange flavors were used. Because fructose is sweeter than galactose, saccharin was added to the galactose solution to increase its palatability. Pre- and posttraining preferences for the galactose and fructose solutions were evaluated in two-bottle choice tests. Also, preferences for the sugar-paired flavors were evaluated in two-bottle tests with the flavors presented in saccharin. In Experiment 1, rats were trained with flavored 80 g/L fructose, 80 g/L galactose + 2 g/L saccharin, and 2 g/L saccharin solutions (20 mL/d). Their preference for the flavored galactose solution changed (P < 0.01) from 76% (pretraining) to 19% (posttraining). The rats also avoided (P < 0.05) the flavor paired with the galactose solution in choice tests with the fructose-paired flavor and the saccharin-paired flavor. Similar pre- to posttraining preference reversals were obtained in Experiments 2 and 3, which used 20 g/L galactose and fructose solutions, and 20 g/L galactose and fructose solutions mixed with 20 g/L glucose, respectively. These findings, together with the intragastric infusion data, demonstrate that galactose has aversive postingestive consequences in adult rats even at low concentrations (20 g/L). Unlike lactose intolerance, which is due to intestinal malabsorption, this galactose-induced flavor avoidance is presumably due to the slow and incomplete postabsorptive metabolism of galactose. PMID- 10460214 TI - Essential amino acids affect interstitial dopamine metabolites in the anterior piriform cortex of rats. AB - The anterior piriform cortex (APC) is essential for the anorectic reactions to an amino acid-imbalanced diet, and it also responds to repletion of the limiting amino acid. In the present study, we examine the dynamic changes of the interstitial dopamine metabolites in the APC following feeding of either an amino acid-corrected or -imbalanced diet. Microdialysates, collected from the APC, were analyzed using HPLC with electrochemical detection. The concentrations were 19.7 +/- 4.8 microg/L for 3, 4-dyhydroxyphenylacetic acid and 25.1 +/- 4.4 microg/L for homovanillic acid, respectively, in the baseline dialysates. After diet treatments, no significant changes occurred in 3, 4-dyhydroxyphenylacetic acid in the corrected (n = 7) or imbalanced (n = 9) groups vs. the basal group (n = 7). However, after feeding the threonine-corrected diet, the concentration of homovanillic acid was significantly less (P < 0.01) than after the basal and imbalanced diets. The homovanillic acid level in the corrected group was already significantly lower than in the basal group by 20 min (P < 0.05), and reached its lowest level at 70 min (P < 0.05). The concentrations of homovanillic acid in the corrected group remained at this low level until the end of the experiment. The present results introduce the idea that the dopaminergic system is involved in the feeding responses to essential amino acid repletion. PMID- 10460215 TI - The decidualizing effect of progesterone may involve direct transcriptional activation of corticotrophin-releasing hormone from human endometrial stromal cells. AB - The hypothalamic neuropeptide corticotrophin-releasing hormone (CRH) is also produced by human endometrial cells and is directly involved in the decidualization process as a paracrine inducer. The aim of the present work was to examine the effect of progesterone, the main decidualizing factor, on endometrial CRH, in primary cultures of human endometrial stromal cells. The effect of progesterone was examined by measuring the effects of medroxyprogesterone acetate (MPA) on (i) the concentration of immunoreactive CRH in isolated human endometrial stromal cells and (ii) the activity of the CRH promoter in human endometrial stromal cells transfected with a 0.9 kb fragment of the 5'-flanking region of the human CRH gene coupled to luciferase reporter. The data show that MPA increased the production and secretion of immunoreactive CRH from stromal cells and induced the activity of the CRH promoter, both in a dose dependent manner. These effects were partially reversed by a molar excess of the antiprogestin RU 486 and were completely abolished in the presence of 100 nmol/l of the cAMP inhibitor, Rp-cAMP. The effect of progesterone on the CRH promoter requires the existence of an intact CRH sequence since experiments carried out with a deleted palindromic cAMP response element (CRE: 5'-TGACGTCA) at -224 bp of the CRH promoter resulted in a complete loss of MPA effect. In conclusion, these data provide evidence that progesterone induces the transcription of CRH gene in human endometrial stroma. This effect coupled with the decidualizing properties of progesterone and CRH may indicate that progesterone and CRH form a decidualizing local pathway within the human endometrium. PMID- 10460216 TI - Association of oestrogen receptor gene polymorphisms with outcome of ovarian stimulation in patients undergoing IVF. AB - Oestrogen plays an important role in follicular formation and oocyte maturation via its receptor (ER). Many studies have shown association of the ER gene polymorphisms with a variety of pathological conditions. In this study we have examined the relationship of a common PvuII and a rare BstUI polymorphism in the ER gene to the mean numbers of follicles and oocytes, their mean ratios, mean number of embryos, mean oestrogen concentrations, mean size of the follicles and pregnancy rates. Analyses were carried out in 200 local Chinese patients undergoing in-vitro fertilization (IVF) and embryo transfer in three consecutive cycles. The mean follicular number, oocyte number, embryo number, follicular size and pregnancy rate were significantly smaller in patients homozygous for PvuII polymorphism (P < 0.001). These results indicate that PvuII polymorphism may be associated with ovarian follicular development and subsequently with the pregnancy rate. This study supports the view that genetic variability in the ER gene may have a role in the quality of the ovarian follicles in stimulation, which may affect implantation. However BstUI polymorphism was not found in either the IVF or control groups, suggesting that it has no role in the local Chinese population. PMID- 10460217 TI - Cyclic expression of mRNA transcripts for connective tissue components in the mouse ovary. AB - In the ovary, differentiation of germinal cells into primordial follicles, functional ovulatory follicles and corpus luteum, all take place in a connective tissue matrix. We postulated that extracellular matrix (ECM) of the ovary participates actively in ovarian functions. To test this, the mRNA levels for several ECM components were determined in the mouse ovary at six distinct stages of the 4-day oestrous cycle. Northern analysis revealed statistically significant cyclic expression patterns for the mRNAs coding for type III, IV and VI collagens as well as for the small proteoglycan, biglycan, and for syndecan-1 and osteonectin. The cyclic changes observed in the mRNAs for these structural components exceeded those for matrix metalloproteinases (MMP)-2, -9 and -13, and for tissue inhibitors of matrix metalloproteinases (TIMP)-1, -2 and -3, where the changes were not statistically significant, despite their apparent role in ECM remodelling in the ovary. These observations support the hypothesis that cyclic changes in the production and degradation of ECM are part of normal ovarian function connected with follicular maturation, rupture and corpus luteum formation. PMID- 10460218 TI - Characterization of human semen alpha-L-fucosidases. AB - Human semen contains a large amount of alpha-L-fucosidase activity, the great majority of which is found in the seminal fluid. Immunocytochemical studies indicate that a small amount of semen fucosidase activity is present on the sperm plasma membrane, primarily in the posterior head region. Subcellular fractionation studies also indicate that sperm alpha-L-fucosidase is present in the plasma membrane-enriched fraction. Comparative characterization of human seminal fluid and sperm alpha-L-fucosidases indicates that seminal fluid alpha-L fucosidase has a broad pH optimum curve with a number of near-equal maxima between pH 4.8 and 7.0 while sperm fucosidase has a major optimum between pH 3.4 and 4.0. Isoelectric focusing indicates that seminal fluid alpha-L-fucosidase contains three to six isoforms with isoelectric points (pI) of 5-7 while sperm fucosidase contains two distinct isoforms with pI values of 5. 2 +/- 0.2 and 7.0 +/- 0.2. Western blotting indicates that seminal fluid fucosidase contains a major protein band with a molecular mass ratio (M(r)) of approximately 56 kDa while sperm fucosidase contains a major protein band of approximately 51 kDa. The overall results indicate the presence of a low-abundance, plasma membrane associated human sperm alpha-L-fucosidase, which is different in its properties from human seminal fluid alpha-L-fucosidase(s), and whose function is not yet known. PMID- 10460219 TI - Relationship between sperm motility and the processing and tyrosine phosphorylation of two human sperm fibrous sheath proteins, pro-hAKAP82 and hAKAP82. AB - Sperm motility is regulated by the cAMP-dependent protein kinase (protein kinase A)-mediated phosphorylation of a group of largely unidentified flagellar proteins. Human AKAP82 (hAKAP82) and its precursor protein, pro-hAKAP82, are members of the A-kinase anchor protein (AKAP) family. These proteins tether protein kinase-A to the fibrous sheath of human spermatozoa and presumably localize the activity of the kinase near specific targets in the sperm flagellum. In this way, pro-hAKAP82 and hAKAP82 may be involved in regulating sperm motility. Similar to its homologues in other species, pro-hAKAP82 is proteolytically processed to hAKAP82. However, the amount of processing of pro hAKAP82 in human spermatozoa is less than the amount of processing of the precursor in other species. We postulated that this lower extent of processing may be related to lower percentages of human sperm motility. In addition, both pro-hAKAP82 and hAKAP82 are tyrosine phosphorylated in a capacitation-dependent manner. Since capacitation is associated with hyperactivated motility, we postulated that tyrosine phosphorylation of pro-hAKAP82/hAKAP82 is associated with changes in motility. However, using a combination of immunofluorescence and immunoblotting approaches, we found no evidence for an association between either processing or tyrosine phosphorylation of pro-hAKAP82/hAKAP82 and significant differences in motility in spermatozoa from normal men. PMID- 10460220 TI - Relationship between abnormal sperm chromatin packing and IVF results. AB - This study was initiated to determine the relationship between the fertilizing potential of spermatozoa and abnormalities in the compact packing of their chromatin which occurs in the final stage of male germ cell differentiation. Chromatin packing involves disulphide bridge covalent cross-linking. The degree of packing was determined from the accessibility of DNA to a fluorescent dye, ethidium bromide, following detergent treatment of the spermatozoa. The amount of dye bound was determined by flow cytometry in the presence or absence of heparin, a polyanion which removes only non-disulphide bridge-linked proteins. The results of a number of different sperm samples were compared with their results following in-vitro fertilization, and a relationship between disordered sperm chromatin packing and rates of embryo cleavage was observed. This study suggests that abnormal chromatin packing in spermatozoa may contribute to male fertility. PMID- 10460221 TI - A comparison of DNA damage in testicular and proximal epididymal spermatozoa in obstructive azoospermia. AB - Testicular and epididymal spermatozoa are used routinely for intracytoplasmic sperm injection (ICSI) to treat men with obstructive azoospermia. Little is known of the effects of obstruction and stasis on the DNA of these spermatozoa, particularly in the epididymis where spermatozoa have been retained for long periods. Surgical epididymal aspiration for ICSI could provide spermatozoa that are senescent or dying. Using the Comet assay, the percentage of undamaged DNA of testicular spermatozoa from 20 men with obstructive azoospermia was significantly better (83.0 +/- 1. 2%) than from proximal epididymal spermatozoa (75.4 +/- 2.3%; P < 0. 05). There was no difference between the percentage of undamaged DNA of testicular spermatozoa from 39 men with obstructive azoospermia (84.0 +/- 0.9) or from 10 fertile men at vasectomy (86.8 +/- 1.8) or from ejaculated spermatozoa from five of the controls (78.9 +/- 3.9; P > 0.05). In nine subjects, a second biopsy was carried out 6 months later. There was no significant difference in undamaged DNA on these two occasions (83.5 +/- 5.6 and 84.1 +/- 4.2; P > 0.05). This confirms the reproducibility of the Comet assay for non-ejaculated spermatozoa. Our data suggest that testicular sperm DNA appears to be significantly less damaged than epididymal sperm DNA, and so testicular spermatozoa should be used in preference for ICSI to treat men with obstructive azoospermia. PMID- 10460222 TI - Sperm integrity is critical for normal mitotic division and early embryonic development. AB - The human zygote relies on the paternal gamete to provide the centrosome component essential for the first mitotic division. It is not known whether normal centrosome function requires an intact spermatozoon, or whether donation of an isolated paternal centrosome component can result in normal zygotes and embryos. To explore this possibility, mature human oocytes were microinjected with either intact or dissected spermatozoa. Fertilization and cleavage rates were documented; nuclear and cytoskeletal changes were observed with fluorescent immunocytochemistry; and chromosomal normality was assessed with fluorescent in situ hybridization. A pilot study was performed to identify cytoskeletal features suggestive of centrosome function. Unfertilized oocytes and tripronucleate (3PN) zygotes from in-vitro fertilization or intracytoplasmic sperm injection were assessed to confirm the sequence of the landmarks of human fertilization. Oocytes injected with mechanically-dissected spermatozoa appear to be capable of normal pronuclear formation and embryonic cleavage, but do not undergo normal mitotic division. Although decondensed, apposed nuclei are noted in combination with diffuse cytoskeleton assembly, no spindle was detected in any zygote resulting from the injection of a dissected spermatozoon. Analysis of selected embryos resulting from dissected sperm injection revealed chromosomal mosaicism in the majority of specimens. The lack of a bipolar spindle, in combination with chromosomal mosaicism, suggests abnormalities of the mitotic apparatus when sperm integrity is impaired following dissection. PMID- 10460223 TI - Alternative splicing of the telomerase catalytic subunit in human oocytes and embryos. AB - The human telomerase catalytic subunit (hTCS) is a ribonucleoprotein which synthesizes telomere repeats on the ends of chromosomes. Telomerase activity is thought to be essential in maintaining normal telomere length in immortal (cancer) and germ cells. The objective of this study was to determine the gene expression of telomerase mRNA in human oocytes at different meiotic stages and in embryos. Normal and abnormal human oocytes, preimplantation embryos, and blastocysts were analysed for the presence and expression of the hTCS transcripts. Multiple telomerase mRNA products were identified by reverse transcription-polymerase chain reaction (RT-PCR) using primers within the reverse transcriptase domain. DNA sequencing of these amplicons suggest that there are alternative splicing variants which align to other telomerase reverse transcriptase (RT) consensus domains. Surprisingly, in unfertilized and immature gametes, as well as preimplantation embryos, hTCS expression revealed three different PCR product sizes, 457, 421 and 275 bp. The frequency of the 275 bp DNA product was 6.6% in oocytes (two out of 30) compared with 56.6% (17 out of 30) in poorly developing human preimplantation embryos (P < 0.005). The presence of alternately spliced mRNA variants in human preimplantation embryos may suggest a lack of telomerase activity and thus chromosomes associated with shortened telomeres. PMID- 10460224 TI - Identification of genes expressed in human primordial germ cells at the time of entry of the female germ line into meiosis. AB - In mammals, primordial germ cells (PGCs) are first observed in the extraembryonic mesoderm from where they migrate through the hindgut and its mesentery to the genital ridge to colonize the developing gonads. Soon after reaching the gonads, the female PGCs enter meiosis, while the male PGCs are arrested in mitosis and enter meiosis postnatally. To gain an insight into the molecular events controlling human germ cell development, we determined specific profiles of gene expression using cDNA prepared from PGCs isolated from male and female fetal gonads at 10 weeks gestation, when female PGCs start to enter meiosis. The identity of the isolated PGCs, and the cDNA molecules prepared from them, was confirmed respectively, by alkaline phosphatase staining and by the presence of transcripts of OCT4, a marker gene for PGCs and pluripotent stem cells in mice. Using differential display to compare the profiles of gene expression of male and female germ cells with each other and with that of a whole 10 week old fetus, we have identified eight transcripts differentially expressed in male and/or female germ cells. Among these transcripts, we have identified a member of the olfactory receptor gene family, which contains genes known to be germline-specific in the dog and possibly associated with chemotactic function. Another transcript is common to a previously isolated sequence from the human testis and we have extended this sequence towards the 5' end for partial characterization. The germline-specific sequences also include two novel sequences not represented in the databases. These findings are highly encouraging for the elucidation of the genetic programming of male and female germ line development. PMID- 10460225 TI - Expression of the insulin-like growth factor-1 gene and its receptor in preimplantation mouse embryos; is it a marker of embryo viability? AB - Recent studies have demonstrated the importance of insulin-like growth factors (IGF) in mouse preimplantation development. We examined IGF-1 and IGF-1 receptor (IGF-1R) gene expression in a single blastomere of an early mouse embryo and compared it with subsequent embryo development in culture. Fertilized eggs and 2 cell embryos were obtained by tubal flushing in superovulated and mated female mice. Single cells were removed from embryos at cleavage stage between 3 and 8 cells using the standard embryo biopsy techniques. Individual blastomeres from each embryo were then assayed for the presence of IGF-1 and IGF-1R mRNA using reverse transcription-polymerase chain reaction. The biopsied embryos were washed in medium and placed in co-culture with murine endometrial cells. Embryonic development in culture was assessed and blastocyst grading was performed. IGF-1 gene expression was then examined for an association with in-vitro development. Eighty-seven embryos were biopsied. IGF-1R gene expression was detected in the majority of embryos tested and IGF-1 gene expression was detected in 34 of 81 (42%) embryos. A significant association between IGF-1 expression and blastocyst formation in vitro was found (P < 0.01). There was no association between IGF-1R expression and subsequent embryo development. We conclude that IGF-1 gene expression could potentially be used as a marker of embryo quality. PMID- 10460226 TI - Regulation of HOXA-10 and its expression in normal and abnormal endometrium. AB - HOXA-10 is a member of a family of genes that serve as transcription factors during development and have been shown to be important for uterine function. Using immunohistochemistry and RNAse protection assays (RPA), HOXA-10 was shown to be expressed in both epithelial and stromal cells with increased expression during the window of implantation. By in-vitro culture of isolated endometrial epithelium or stroma, HOXA-10, expression was increased after treatment with oestradiol (10(-8) mol/l) with or without progesterone (10(-6) mol/l). In stromal cells, oestradiol and progesterone both appeared to increase HOXA-10 expression and were additive. Relaxin (30 ng/ml) appeared to further increase stromal HOXA 10 expression. HOXA-10 expression during the window of implantation was compared in normal menstrual cycles to endometrium from women with endometriosis and suspected defects in uterine receptivity. Little or no difference was seen in luminal, glandular or endothelial HOXA-10 expression but a significant reduction in stroma HOXA-10 expression was noted in women with endometriosis. In conclusion, HOXA-10 is a hormone-regulated endometrial transcription factor that appears to be responsive to both ovarian steroids and relaxin. The appearance of this nuclear protein during the window of implantation in epithelium and stroma may offer new insight into the regulation of uterine receptivity and assist in the identification of other genes that are critical to the establishment of a successful pregnancy. PMID- 10460227 TI - The potential role of stem cell factor and its receptor c-kit in the mouse blastocyst implantation. AB - Embryo implantation is a complex process that requires the interaction of embryo and endometrium. Several growth factors and cytokines appear to be involved in this process. Stem cell factor (SCF) and its receptor c-kit regulate the proliferation and survival of germ cells and play an important role in follicular development. However, little information is available on the role of SCF and c kit in the process of blastocyst implantation. In the present study, we examined the expression of SCF and c-kit mRNA in mouse embryos and in the stromal and epithelial cells of the uterine endometrium by reverse transcription-polymerase chain reaction (RT-PCR). SCF mRNA was expressed in the spreading blastocysts and endometrial cells, with especially strong expression occurring in the stromal cells. Expression of c-kit mRNA was detected in the blastocysts and spreading blastocysts, as well as in the endometrial cells. By immunocytochemical studies, staining for c-kit protein was observed in the in-vitro spreading trophoblasts. We found that 50-100 ng/ml SCF significantly promoted the expansion of the surface area of the spreading blastocysts (P < 0.01). These results are consistent with the hypothesis that SCF derived from endometrial cells and the implanting embryo exerts paracrine and/or autocrine action on the process of implantation by stimulating trophoblast outgrowth through its receptor c-kit. PMID- 10460228 TI - Expression of cyclo-oxygenase types-1 and -2 in human myometrium throughout pregnancy. AB - Human labour is associated with increased prostaglandin synthesis within the uterus. The aim of this study was to examine the expression of the two isoforms of the central prostaglandin synthetic enzyme, cyclo-oxygenase (COX-1 and COX-2) in human myometrium throughout pregnancy and to test the hypothesis that COX in the myometrium may play a role in labour onset. Expression of COX-1 and COX-2 at the mRNA level was analysed using reverse transcriptase-polymerase chain reaction (RT-PCR) and at the protein level using Western blotting. No significant changes of COX-1 RNA or protein expression were observed either with gestational age or labour. COX-2 mRNA and protein expression increased at term with significant up regulation occurring prior to the onset of labour (P < 0.005). These data would suggest that up-regulation of COX-2, rather than COX-1, mediates increased prostaglandin synthesis in human myometrium at term. The increased COX-2 expression observed preceded labour onset, suggesting that COX-2 has a role in labour onset, rather than its presence merely a consequence of labour. PMID- 10460229 TI - Human placental cells show enhanced production of interleukin (IL)-8 in response to lipopolysaccharide (LPS), IL-1 and tumour necrosis factor (TNF)-alpha, but not to IL-6. AB - Interleukin-8 (IL-8) is a chemotactic and activating factor for neutrophils which play important roles in host defence mechanisms. The human placenta constitutively produces IL-8 during pregnancy and enhances its production in chorioamnionitis. The present study was designed to investigate in vitro the regulatory mechanism for IL-8 production in the placentas in normal and inflammatory states. Placental cells produced IL-8 in a dose-dependent fashion when stimulated with lipopolysaccharide (LPS). The purified trophoblasts showed significantly higher IL-8 production than untreated placental cells. The expression of IL-8 gene in the trophoblasts in the third trimester was observed by reverse transcription-polymerase chain reaction (RT-PCR). The placental cells also release IL-8 in a dose-dependent manner, in response to r-(recombinant) IL 1alpha and tumour necrosis factor (TNF)-alpha, but not rIL-6. Moreover, LPS activated placental cells spontaneously produced a much larger amount of IL-8 and showed increased responses to rIL-1alpha and TNF-alpha. It may, therefore, be proposed that placental cells with multiple endocrine functions exert immunological functions by constitutive production of IL-1 and TNF-alpha, which stimulate placental IL-8 release. This cytokine cascade in the placenta may be augmented by LPS in chorioamnionitis, thereby potentiating the feto-maternal defence mechanisms against infection. PMID- 10460230 TI - Reactive oxygen species mediate activity-dependent neuron-glia signaling in output fibers of the hippocampus. AB - Nonsynaptic signaling is becoming increasingly appreciated in studies of activity dependent changes in the nervous system. We investigated the types of neuronal activity that elicit nonsynaptic communication between neurons and glial cells in hippocampal output fibers. High-frequency, but not low-frequency, action potential firing in myelinated CA1 axons of the hippocampus resulted in increased phosphorylation of the oligodendrocyte-specific protein myelin basic protein (MBP). This change was blocked by tetrodotoxin, indicating that axonally generated action potentials were necessary to regulate the phosphorylation state of MBP. Furthermore, scavengers of the reactive oxygen species superoxide and hydrogen peroxide and nitric oxide synthase inhibitors prevented activation of this neuron-glia signaling pathway. These results indicate that, during periods of increased neuronal activity in area CA1 of the hippocampus, reactive oxygen and nitrogen species are generated, which diffuse to neighboring oligodendrocytes and result in post-translational modifications of MBP, a key structural protein in myelin. Thus, in addition to their well-known capacity for activity-dependent neuron-neuron signaling, hippocampal pyramidal neurons possess a mechanism for activity-dependent neuron-glia signaling. PMID- 10460231 TI - Caffeine-sensitive calcium stores regulate synaptic transmission from retinal rod photoreceptors. AB - We investigated the role of caffeine-sensitive intracellular stores in regulating intracellular calcium ([Ca(2+)](i)) and glutamatergic synaptic transmission from rod photoreceptors. Caffeine transiently elevated and then markedly depressed [Ca(2+)](i) to below prestimulus levels in rod inner segments and synaptic terminals. Concomitant with the depression was a reduction of glutamate release and a hyperpolarization of horizontal cells, neurons postsynaptic to rods. Caffeine did not affect the rods' membrane potentials indicating that caffeine likely acted via some mechanism(s) other than a voltage-dependent deactivation of the calcium channels. Most of caffeine's depressive action on [Ca(2+)](i), on glutamate release, and on I(Ca) in rods can be attributed to calcium release from stores: (1) caffeine's actions on [Ca(2+)](i) and I(Ca) were reduced by intracellular BAPTA and barium substitution for calcium, (2) other nonxanthine store-releasing compounds, such as thymol and chlorocresol, also depressed [Ca(2+)](i), and (3) the magnitude of [Ca(2+)](i) depression depended on basal [Ca(2+)](i) before caffeine. We propose that caffeine-released calcium reduces I(Ca) in rods by an as yet unidentified intracellular signaling mechanism. To account for the depression of [Ca(2+)](i) below rest levels and the increased fall rate of [Ca(2+)](i) with higher basal calcium, we also propose that caffeine evoked calcium release from stores activates a calcium transporter that, via sequestration into stores or extrusion, lowers [Ca(2+)](i) and suppresses glutamate release. The effects of store-released calcium reported here operate at physiological calcium concentrations, supporting a role in regulating synaptic signaling in vivo. PMID- 10460232 TI - Presynaptic mechanism for phorbol ester-induced synaptic potentiation. AB - Phorbol ester facilitates transmitter release at a variety of synapses, and the phorbol ester-induced synaptic potentiation (PESP) is a model for presynaptic facilitation. To address the mechanism underlying PESP, we have made paired whole cell recordings from the giant presynaptic terminal, the calyx of Held, and its postsynaptic target in the medial nucleus of the trapezoid body in rat brainstem slices. Phorbol ester potentiated EPSCs without affecting either presynaptic calcium currents or potassium currents. Protein kinase C inhibitors applied from outside or injected directly into the presynaptic terminal attenuated the PESP. Furthermore, presynaptic loading of a synthetic peptide with the sequence of the N-terminal domain of Doc2alpha interacting with Munc13-1 (Mid peptide) significantly attenuated PESP, whereas mutated Mid peptide had no effect. We conclude that the target of the presynaptic facilitatory effect of phorbol ester resides downstream of calcium influx and may involve both protein kinase C and Doc2alpha - Munc13-1 interaction. PMID- 10460233 TI - Properties of Q-type calcium channels in neostriatal and cortical neurons are correlated with beta subunit expression. AB - In brain neurons, P- and Q-type Ca(2+) channels both appear to include a class A alpha1 subunit. In spite of this similarity, these channels differ pharmacologically and biophysically, particularly in inactivation kinetics. The molecular basis for this difference is unclear. In heterologous systems, alternative splicing and ancillary beta subunits have been shown to alter biophysical properties of channels containing a class A alpha1 subunit. To test the hypothesis that similar mechanisms are at work in native systems, P- and Q type currents were characterized in acutely isolated rat neostriatal, medium spiny neurons and cortical pyramidal neurons using whole-cell voltage-clamp techniques. Cells were subsequently aspirated and subjected to single-cell RT-PCR (scRT-PCR) analysis of calcium channel alpha(1) and beta (beta(1-4)) subunit expression. In both cortical and neostriatal neurons, P- and Q-type currents were found in cells expressing class A alpha(1) subunit mRNA. Although P-type currents in cortical and neostriatal neurons were similar, Q-type currents differed significantly in inactivation kinetics. Notably, Q-type currents in neostriatal neurons were similar to P-type currents in inactivation rate. The variation in Q type channel biophysics was correlated with beta subunit expression. Neostriatal neurons expressed significantly higher levels of beta(2a) mRNA and lower levels of beta(1b) mRNA than cortical neurons. These findings are consistent with the association of beta(2a) and beta(1b) subunits with slow and fast inactivation, respectively. Analysis of alpha(1A) splice variants in the linker between domains I and II failed to provide an alternative explanation for the differences in inactivation rates. These findings are consistent with the hypothesis that the biophysical properties of Q-type channels are governed by beta subunit isoforms and are separable from toxin sensitivity. PMID- 10460234 TI - Formation of compact myelin is required for maturation of the axonal cytoskeleton. AB - Although traditional roles ascribed to myelinating glial cells are structural and supportive, the importance of compact myelin for proper functioning of the nervous system can be inferred from mutations in myelin proteins and neuropathologies associated with loss of myelin. Myelinating Schwann cells are known to affect local properties of peripheral axons (de Waegh et al., 1992), but little is known about effects of oligodendrocytes on CNS axons. The shiverer mutant mouse has a deletion in the myelin basic protein gene that eliminates compact myelin in the CNS. In shiverer mice, both local axonal features like phosphorylation of cytoskeletal proteins and neuronal perikaryon functions like cytoskeletal gene expression are altered. This leads to changes in the organization and composition of the axonal cytoskeleton in shiverer unmyelinated axons relative to age-matched wild-type myelinated fibers, although connectivity and patterns of neuronal activity are comparable. Remarkably, transgenic shiverer mice with thin myelin sheaths display an intermediate phenotype indicating that CNS neurons are sensitive to myelin sheath thickness. These results indicate that formation of a normal compact myelin sheath is required for normal maturation of the neuronal cytoskeleton in large CNS neurons. PMID- 10460235 TI - Allosteric control of gating and kinetics at P2X(4) receptor channels. AB - The CNS abundantly expresses P2X receptor channels for ATP; of these the most widespread in the brain is the P2X(4) channel. We show that ivermectin (IVM) is a specific positive allosteric effector of heterologously expressed P2X(4) and possibly of heteromeric P2X(4)/P2X(6) channels, but not of P2X(2), P2X(3), P2X(2)/P2X(3,) or P2X(7) channels. In the submicromolar range (EC(50,) approximately 250 nM) the action of IVM was rapid and reversible, resulting in increased amplitude and slowed deactivation of P2X(4) channel currents evoked by ATP. IVM also markedly increased the potency of ATP and that of the normally low potency agonist alpha, beta-methylene-ATP in a use- and voltage-independent manner without changing the ion selectivity of P2X(4) channels. Therefore, IVM evokes a potent pharmacological gain-of-function phenotype that is specific for P2X(4) channels. We also tested whether IVM could modulate endogenously expressed P2X channels in the adult trigeminal mesencephalic nucleus and hippocampal CA1 neurons. Surprisingly, IVM produced no significant effect on the fast ATP-evoked inward currents in either type of neuron, despite the fact that IVM modulated P2X(4) channels heterologously expressed in embryonic hippocampal neurons. These results suggest that homomeric P2X(4) channels are not the primary subtype of P2X receptor in the adult trigeminal mesencephalic nucleus and in hippocampal CA1 neurons. PMID- 10460236 TI - The insulin receptor tyrosine kinase substrate p58/53 and the insulin receptor are components of CNS synapses. AB - The synapse is the primary locus of cell-cell communication in the nervous system. It is now clear that the synapse incorporates diverse cell signaling modalities in addition to classical neurotransmission. Here we show that two components of the insulin pathway are localized at CNS synapses, where they are components of the postsynaptic density (PSD). An immunochemical screen revealed that polypeptides of 58 and 53 kDa (p58/53) were highly enriched in PSD fractions from rat cerebral cortex, hippocampus, and cerebellum. These polypeptides were purified and microsequenced, revealing that p58/53 is identical to the insulin receptor tyrosine kinase substrate p58/53 (IRSp53). Our analysis of IRSp58/53 mRNA suggests that within rat brain there is one coding region for IRSp58 and IRSp53; we find no evidence of alternative splicing. We demonstrate that IRSp58/53 is expressed in the synapse-rich molecular layer of the cerebellum and is highly concentrated at the synapses of cultured hippocampal neurons, where it co-localizes with the insulin receptor. Together, these data suggest that insulin signaling may play a role at CNS synapses. PMID- 10460237 TI - Ionic basis for plateau potentials in deep dorsal horn neurons of the rat spinal cord. AB - Approximately 28% of dorsal horn neurons (DHNs) in lamina V of the rat spinal cord generate voltage-dependent plateau potentials underlying accelerating discharges and prolonged afterdischarges in response to steady current pulses or stimulation of nociceptive primary afferent fibers. Using intracellular recordings in a transverse slice preparation of the cervical spinal cord, we have analyzed the ionic mechanisms involved in the generation and maintenance of plateau potentials in lamina V DHNs. Both the accelerating discharges and afterdischarges were reversibly blocked by Mn(2+) and enhanced when Ca(2+) was substituted with Ba(2+). The underlying tetrodotoxin-resistant regenerative depolarization was sensitive to dihydropyridines, being blocked by nifedipine and enhanced by Bay K 8644. Substitution of extracellular Na(+) with N-methyl-D glucamine or choline strongly decreased the duration of the plateau potential. Loading the neurons with the calcium chelator BAPTA did not change the initial response but clearly decreased the maximum firing frequency and the duration of the afterdischarge. A similar effect was obtained with flufenamate, a specific blocker of the calcium-activated nonspecific cation current (I(CAN)). We conclude that the plateau potential of deep DHNs is supported by both Ca(2+) influx through intermediate-threshold voltage-gated calcium channels of the L-type and by subsequent activation of a CAN current. Ca(2+) influx during the plateau is potentially of importance for pain integration and the associated sensitization in spinal cord. PMID- 10460238 TI - A neuronal-specific mammalian homolog of the Drosophila retinal degeneration B gene with expression restricted to the retina and dentate gyrus. AB - Mutations in the Drosophila retinal degeneration B (rdgB) gene cause a rapid loss of the electrophysiological light response and subsequent light-enhanced photoreceptor degeneration. The rdgB gene encodes a protein with an N-terminal phosphatidylinositol transfer protein domain, a large C-terminal segment, and several hydrophobic regions thought to multiply span the subrhabdomeric cisternal membrane. A mammalian rdgB homolog (m-rdgB1) was previously identified and shown to exhibit widespread tissue distribution and functionally rescue the Drosophila rdgB mutant phenotypes. We describe a second mammalian rdgB homolog (m-rdgB2) that possesses 46% amino acid identity to Drosophila RdgB and 56% identity to M RdgB1. M-RdgB2 possesses a neuronal-specific expression pattern, with high levels in the retina and the dentate gyrus mossy fibers and dendritic field. Using M RdgB2-specific antibodies and subcellular fractionation, we demonstrate that M RdgB2 is not an integral membrane protein but is stably associated with a particulate fraction through protein-protein interactions. Although transgenic expression of M-RdgB2 in rdgB2 null mutant flies suppressed the retinal degeneration, it failed to fully restore the electrophysiological light response. Because transgenic expression of M-RdgB2 does not restore the wild-type phenotype to rdgB2 mutant flies to the same extent as M-RdgB1, functional differences likely exist between the two M-RdgB homologs. PMID- 10460239 TI - Entire course and distinct phases of day-lasting depression of miniature EPSC amplitudes in cultured Purkinje neurons. AB - The cerebellar long-term depression (LTD) is the long-lasting reduction of transmission efficacy at the granule neuron-Purkinje neuron (G-P) synapses and is a candidate mechanism for the motor learning. Despite extensive studies on its induction and expression mechanisms, it has not been known how long the LTD lasts. The LTD is accompanied by the decrease in the postsynaptic responsiveness to glutamate, the transmitter at G-P synapses. Therefore, during the LTD, the amplitude of miniature EPSCs (mEPSCs) at G-P synapses should decrease. We studied the depression of mEPSC amplitudes (DME) as a possible contributing factor for the LTD and found that the conditioning treatment of cultured cerebellar neurons with 50 mM K(+) and 100 microM glutamate, an analogous condition used to induce the LTD, induced the long-lasting DME. The mEPSC amplitudes recovered to the original level 48 hr after the 5 min conditioning treatment. Changing the duration of the conditioning revealed that the DME consisted of two distinct phases: the early phase lasting for a few hours and the late phase for >1 d. The latter was distinguished from the former by its requirement of prolonged conditioning treatment and syntheses of mRNA and protein for the induction. There were critical periods for mRNA and protein syntheses. The critical period for protein synthesis was much longer than that for mRNA synthesis. These results demonstrate that the DME lasts for 1-2 d and that it consists of two phases, whose induction and maintenance mechanisms are distinct. PMID- 10460240 TI - High conductance sustained single-channel activity responsible for the low threshold persistent Na(+) current in entorhinal cortex neurons. AB - Stellate cells from entorhinal cortex (EC) layer II express both a transient Na(+) current (I(Na)) and a low-threshold persistent Na(+) current (I(NaP)) that helps to generate intrinsic theta-like oscillatory activity. We have used single channel patch-clamp recording to investigate the Na(+) channels responsible for I(NaP) in EC stellate cells. Macropatch (more than six channels) recordings showed high levels of transient Na(+) channel activity, consisting of brief openings near the beginning of depolarizing pulses, and lower levels of persistent Na(+) channel activity, characterized by prolonged openings throughout 500 msec long depolarizations. The persistent activity contributed a noninactivating component to averaged macropatch recordings that was comparable with whole-cell I(NaP) in both voltage dependence of activation (10 mV negative to the transient current) and amplitude (1% of the transient current at -20 mV). In 14 oligochannel (less than six channels) patches, the ratio of transient to persistent channel activity varied from patch to patch, with 10 patches exhibiting exclusively transient openings and one patch showing exclusively persistent openings. In two patches containing only a single persistent channel, prolonged openings were observed in >50% of test depolarizations. Moreover, persistent openings had a significantly higher single-channel conductance (19.7 pS) than transient openings (15.6 pS). We conclude that this stable high conductance persistent channel activity is responsible for I(NaP) in EC stellate cells. This persistent channel behavior is more enduring and has a higher conductance than the infrequent and short-lived transitions to persistent gating modes that have been described previously in brain neurons. PMID- 10460241 TI - Junctional versus extrajunctional glycine and GABA(A) receptor-mediated IPSCs in identified lamina I neurons of the adult rat spinal cord. AB - Colocalization of GABA and glycine in synaptic terminals of the superficial dorsal horn raises the question of their relative contribution to inhibition of different classes of neurons in this area. To address this issue, miniature IPSCs (mIPSCs) mediated via GABA(A) receptors (GABA(A)Rs) and glycine receptors (GlyRs) were recorded from identified laminae I-II neurons in adult rat spinal cord slices. GABA(A)R-mediated mIPSCs had similar amplitude and rise times, but significantly slower decay kinetics than GlyR-mediated mIPSCs. Lamina I neurons appeared to receive almost exclusively GlyR-mediated mIPSCs, even after application of hypertonic solutions. Yet, all neurons responded to exogenous applications of both GABA and glycine, indicating that they expressed both GABA(A)Rs and GlyRs. Given that virtually all glycinergic interneurons also contain GABA, the possibility was examined that GABA(A)Rs may be located extrasynaptically in lamina I neurons. A slow GABA(A)R-mediated component was revealed in large, but not minimally evoked monosynaptic IPSCs. Administration of the benzodiazepine flunitrazepam unmasked a GABA(A)R component to most mIPSCs, suggesting that both transmitters were released from the same vesicle. The isolated GABA(A)R component of these mIPSCs had rising kinetics 10 times slower than that of the GlyR component (or of GABA(A)R mIPSCs in lamina II). The slow GABA(A)R components were prolonged by GABA uptake blockers. It is concluded that, whereas GABA and glycine are likely released from the same vesicle of transmitter in lamina I, GABA(A)Rs appear to be located extrasynaptically. Thus, glycine mediates most of the tonic inhibition at these synapses. This differential distribution of GABA(A)Rs and GlyRs confers distinct functional properties to inhibition mediated by these two transmitters in lamina I. PMID- 10460242 TI - Ultrastructural localization of the serotonin transporter in limbic and motor compartments of the nucleus accumbens. AB - Extracellular levels of serotonin [5-hydroxytryptamine (5-HT)] in the nucleus accumbens (NAc) can influence both cognitive and motor functions involving extensive connections with the frontal cortex. The 5-HT levels reflect vesicular release and plasmalemmal reuptake through the serotonin transporter (SERT). We used electron microscopic immunocytochemistry to determine the sites for SERT activation in the limbic shell and motor-associated core of the rat NAc. Of the SERT-immunoreactive profiles in each region, >90% were serotonergic axons and axon terminals; the remainder were nonserotonergic dendrites and glia. Axonal SERT immunogold labeling was seen mainly at nonsynaptic sites on plasma membranes and often near 5-HT-containing large dense core vesicles (DCVs). SERT-labeled axonal profiles were larger and had a higher numerical density in the shell versus the core but showed no regional differences in their content of SERT immunogold particles. In contrast, immunoreactive dendrites had a lower numerical density in the shell than in the core. SERT labeling in dendrites was localized to segments of plasma membrane near synaptic contacts from unlabeled terminals and/or dendritic appositions. Our results suggest that in the NAc (1) reuptake into serotonergic axons is most efficient after exocytotic release from DCVs, and (2) increased 5-HT release without concomitant increase in SERT expression in individual axons may contribute to higher extracellular levels of serotonin in the shell versus the core. These findings also indicate that SERT may play a minor substrate-dependent role in serotonin uptake or channel activity in selective nonserotonergic neurons and glia in the NAc. PMID- 10460243 TI - Enhancement of AMPA-mediated current after traumatic injury in cortical neurons. AB - Overactivation of ionotropic glutamate receptors has been implicated in the pathophysiology of traumatic brain injury. Using an in vitro cell injury model, we examined the effects of stretch-induced traumatic injury on the AMPA subtype of ionotropic glutamate receptors in cultured neonatal cortical neurons. Recordings made using the whole-cell patch-clamp technique revealed that a subpopulation of injured neurons exhibited an increased current in response to AMPA. The current-voltage relationship of these injured neurons showed an increased slope conductance but no change in reversal potential compared with uninjured neurons. Additionally, the EC(50) values of uninjured and injured neurons were nearly identical. Thus, current potentiation was not caused by changes in the voltage-dependence, ion selectivity, or apparent agonist affinity of the AMPA channel. AMPA-elicited current could also be fully inhibited by the application of selective AMPA receptor antagonists, thereby excluding the possibility that current potentiation in injured neurons was caused by the activation of other, nondesensitizing receptors. The difference in current densities between control and injured neurons was abolished when AMPA receptor desensitization was inhibited by the coapplication of AMPA and cyclothiazide or by the use of kainate as an agonist, suggesting that mechanical injury alters AMPA receptor desensitization. Reduction of AMPA receptor desensitization after brain injury would be expected to further exacerbate the effects of increased postinjury extracellular glutamate and contribute to trauma-related cell loss and dysfunctional synaptic information processing. PMID- 10460244 TI - Ca(2+)-dependent activator protein for secretion is critical for the fusion of dense-core vesicles with the membrane in calf adrenal chromaffin cells. AB - Calcium-dependent activator protein for secretion (CAPS) is a neural/endocrine cell-specific protein that has been shown to function at the Ca(2+)-dependent triggering step of dense-core vesicle (DCV) exocytosis in permeabilized PC12 cells. To evaluate the function of CAPS under physiological conditions, we introduced affinity-purified anti-CAPS IgGs into calf adrenal chromaffin (AC) cells via a patch pipette and tested the kinetics of catecholamine secretion using both amperometric and membrane capacitance techniques. The antibodies reacted with a single major approximately 145 kDa protein in AC cells based on immunoblot analysis. AC cells stimulated with sequential trains of action potentials at 7 Hz resulted in successive secretory episodes of equivalent magnitude. When either of two different anti-CAPS IgGs or their Fab fragments were present, a rapid and progressive inhibition of catecholamine release ensued to a maximum of >80%. The effect was specific because preabsorption of IgGs with the respective antigens ablated the inhibitory effect, and the IgGs had no effect on Ca currents. CAPS immunoneutralization not only reduced the number of amperometric spikes but markedly altered the kinetic characteristics of the residual events. The remaining spikes were much smaller (by 85%) and broader (by approximately 3.5-fold) than those in control cells, suggesting that CAPS plays a role in determining release of vesicle contents via the fusion pore. Anti-CAPS IgGs also slowed the rate of the initial exocytotic capacitance burst, representing the docked-and-primed vesicle pool, by approximately 90% but had no effect on the kinetics of rapid endocytosis. These results suggest that CAPS is a key component regulating the fusion of DCVs to the plasma membrane, and possibly fusion pore dilation, in catecholamine secretion from AC cells. PMID- 10460245 TI - Evidence of an agrin receptor in cortical neurons. AB - Agrin plays a key role in directing the differentiation of the vertebrate neuromuscular junction. Understanding agrin function at the neuromuscular junction has come via molecular genetic analyses of agrin as well as identification of its receptor and associated signal transduction pathways. Agrin is also expressed by many populations of neurons in brain, but its role remains unknown. Here we show, in cultured cortical neurons, that agrin induces expression of the immediate early gene c-fos in a concentration-dependent and saturable manner, as expected for a signal transduction pathway activated by a cell surface receptor. Agrin is active in cortical neurons at picomolar concentrations, is Ca(2+) dependent, and is inhibited by heparin and staurosporine. Despite marked differences in acetylcholine receptor (AChR) clustering activity, all alternatively spliced forms of agrin are equally potent inducers of c-fos in cortical neurons. A similar, isoform-independent response to agrin was also observed in cultures prepared from the hippocampus and cerebellum. Only agrin with high AChR-clustering activity was effective in cultured muscle, whereas non-neuronal cells were agrin insensitive. Although consistent with a receptor tyrosine kinase model similar to the muscle-specific kinase-myotube associated specificity component complex in muscle, our data suggest that CNS neurons express a unique agrin receptor. Evidence that neuronal signal transduction is mediated via an increase in intracellular Ca(2+) means that agrin is well situated to influence important Ca(2+)-dependent functions in brain, including neuronal growth, differentiation, and adaptive changes in gene expression associated with synaptic remodeling. PMID- 10460246 TI - Mitochondrial depolarization is not required for neuronal apoptosis. AB - Mitochondria are sites of cellular energy production but may also influence life and death decisions by initiating or inhibiting cell death. Mitochondrial depolarization and the subsequent release of pro-apoptotic factors have been suggested to be required for the activation of a cell death program in some forms of neuronal apoptosis. We induced apoptosis in cultured rat hippocampal neurons by exposure to the protein kinase inhibitor staurosporine (STS) (300 nM). The time course of mitochondrial membrane potential (DeltaPsi(m)) during apoptosis was examined using the probe tetramethylrhodamine ethyl ester (TMRE). Cells exhibited no decrease in TMRE fluorescence, indicative of mitochondrial depolarization, up to 8 hr after STS exposure. Rather, baseline TMRE fluorescence remained unchanged up to 2 hr and thereafter actually increased significantly. Throughout this time period, the mitochondria could also be depolarized with the protonophore carbonyl cyanide p-trifluoromethoxy-phenylhydrazone (FCCP, 0.1 microM), exhibiting the same relative magnitude of fluorescence release (unquenching) as controls. Even after 16 hr of staurosporine treatment, neurons that showed signs of nuclear apoptosis maintained DeltaPsi(m) and could be depolarized with FCCP. In contrast, caspase-3-like activity had increased roughly sevenfold by 2 hr and >20-fold by 8 hr. Double-labeling of hippocampal neurons with the potential-sensitive probe Mitotracker Red Chloromethyl X-Rosamine and an antibody to cytochrome c demonstrated at the subcellular level that mitochondrial cytochrome c release also occurred in the absence of mitochondrial depolarization. These data suggest that mitochondrial depolarization is not a decisive event in neuronal apoptosis. PMID- 10460247 TI - Neutralizing intraspinal nerve growth factor blocks autonomic dysreflexia caused by spinal cord injury. AB - Autonomic dysreflexia is a condition that develops after spinal cord injury in which potentially life-threatening episodic hypertension is triggered by stimulation of sensory nerves in the body below the site of injury. Central sprouting of small-diameter primary afferent fibers in the dorsal horn of the spinal cord occurs concurrently with the development of this condition. We propose a model for the development of autonomic dysreflexia in which increased nerve growth factor (NGF) in the injured cord stimulates small-diameter primary afferent fiber sprouting, thereby magnifying spinal sympathetic reflexes and promoting dysreflexia. We identified this population of afferent neurons using immunocytochemistry for calcitonin gene-related peptide. Blocking intraspinal NGF with an intrathecally-delivered neutralizing antibody to NGF prevented small diameter afferent sprouting in rats 2 weeks after a high thoracic spinal cord transection. In the same rats, this anti-NGF antibody treatment significantly decreased (by 43%) the hypertension induced by colon stimulation. The extent of small-diameter afferent sprouting after cord transection correlated significantly with the magnitude of increases in arterial pressure during the autonomic dysreflexia. Neutralizing NGF in the spinal cord is a promising strategy to minimize the life-threatening autonomic dysreflexia that develops after spinal cord injury. PMID- 10460248 TI - Regulated expression and subcellular localization of syndecan heparan sulfate proteoglycans and the syndecan-binding protein CASK/LIN-2 during rat brain development. AB - The syndecan family of cell surface heparan sulfate proteoglycans interacts via their cytoplasmic C-terminal tail with the PDZ domain of CASK/LIN-2, a membrane associated guanylate kinase homolog. The syndecan-CASK interaction may be involved in intercellular signaling and/or cell adhesion. Here we show that syndecan-1 to syndecan-4 have distinctive mRNA distributions in adult rat brain by in situ hybridization, with syndecan-2 and -3 being the major syndecans expressed in neurons of the forebrain. At the protein level, syndecan-2 and -3 are differentially localized within neurons; syndecan-3 is concentrated in axons, whereas syndecan-2 is localized in synapses. The synaptic accumulation of syndecan-2 occurs late in synapse development. CASK is a cytoplasmic-binding partner for syndecans, and its subcellular distribution changes strikingly during development, shifting from a primarily axonal distribution in the first 2 postnatal weeks to a somatodendritic distribution in adult brain. This change in CASK distribution correlates temporally and spatially with the expression patterns of syndecan-3 and -2, consistent with the association of both of these syndecans with CASK in vivo. In support of this, we were able to coimmunoprecipitate a complex of CASK and syndecan-3 from brain extracts. Our results indicate that specific syndecans are differentially expressed in various cell types of the brain and are targeted to distinct subcellular compartments in neurons, where they may serve specialized functions. Moreover, CASK is appropriately expressed and localized to interact with both syndecan-2 and -3 in different compartments of the neuron throughout postnatal development. PMID- 10460249 TI - Specificity and sensitivity of a human olfactory receptor functionally expressed in human embryonic kidney 293 cells and Xenopus Laevis oocytes. AB - Here, we provide the first evidence for functional expression of a human olfactory receptor protein (OR17-40) and show that recombinant olfactory receptors can be functionally expressed in heterologous systems. A mixture of 100 different odorants (Henkel 100) elicited a transient increase in intracellular [Ca(2+)] in human embryonic kidney 293 (HEK293) cells stably or transiently transfected with the plasmid pOR17-40. By subdividing the odorant mixture into progressively smaller groups, we identified a single component that represented the only effective substance: helional. Only the structurally closely related molecule heliotroplyacetone also activated the receptor. Other compounds, including piperonal, safrole, and vanillin, were completely ineffective. Mock transfected cells and cells transfected with other receptors showed no change in intracellular [Ca(2+)] in response to odor stimulation. We were also able to functionally express OR17-40 in Xenopus laevis oocytes. Coexpression of a "reporter" channel allowed measurement of the response of oocytes injected with the cRNA of the human receptor to the odor mixture Henkel 100. The effective substances were the same (helional, heliotropylacetone) as those identified by functionally expressing the receptor in HEK293 cells and were active at the same, lower micromolar concentration. These findings open the possibility of now characterizing the sensitivity and specificity of many, if not all, of the hundreds of different human olfactory receptors. PMID- 10460250 TI - Synaptic control of glycine and GABA(A) receptors and gephyrin expression in cultured motoneurons. AB - We have evaluated the influence of the secretory phenotype of presynaptic boutons on the accumulation of postsynaptic glycine receptors (GlyRs), type A GABA receptors (GABA(A)Rs), and gephyrin clusters. The cellular distribution of these components was analyzed on motoneurons cultured either alone or with glycinergic and/or GABAergic neurons. In motoneurons cultured alone, we observed gephyrin clusters at nonsynaptic sites and in front of cholinergic boutons, whereas glycine and GABA(A) receptors formed nonsynaptic clusters. These receptors are functionally and pharmacologically similar to those found in cultures of all spinal neurons. Motoneurons receiving GABAergic innervation from dorsal root ganglia neurons displayed postsynaptic clusters of gephyrin and GABA(A)Rbeta but not of GlyRalpha/beta subunits. In motoneurons receiving glycinergic and GABAergic innervation from spinal interneurons, gephyrin, GlyRalpha/beta, and GABA(A)Rbeta formed mosaics at synaptic loci. These results indicate that (1) the transmitter phenotype of the presynaptic element determines the postsynaptic accumulation of specific receptors but not of gephyrin and (2) the postsynaptic accumulation of gephyrin alone cannot account for the formation of GlyR-rich microdomains. PMID- 10460251 TI - Pulsed laser imaging of Ca(2+) influx in a neuroendocrine terminal. AB - The surge of Ca(2+) that triggers vesicle fusion is shaped by the distribution of Ca(2+) channels and the physical relationship between those channels and the exocytotic apparatus. Although channels and the release apparatus are thought to be tightly associated at fast synapses, the arrangement at neuroendocrine cells is less clear. The distribution of Ca(2+) influx near release sites is difficult to determine because of spatial and temporal limitations on Ca(2+) imaging techniques. We now present spatially resolved images of Ca(2+) influx into rat neuroendocrine terminals on a millisecond time scale. Images of voltage-dependent Ca(2+) influx into neurohypophysial terminals were captured after excitation of Ca(2+)-sensitive dyes with pulses of laser light lasting a fraction of a microsecond. Submembranous Ca(2+) increases were detected during the first millisecond of an evoked Ca(2+) tail current. Steep gradients of Ca(2+) were evident, with concentrations near the membrane reaching above 1 microM during a 30 msec depolarization. Ca(2+) influx appeared evenly distributed, even when diffusion was restricted with an exogenous Ca(2+) chelator. During longer depolarizations, mean and peak Ca(2+) concentrations reached an asymptote in parallel, suggesting that Ca(2+) binding proteins near the membrane rapidly buffer Ca(2+) and do not become saturated during prolonged influx. These data support the hypothesis that exocytosis is activated in these terminals by the summation of influx through multiple, randomly spaced Ca(2+) channels. PMID- 10460252 TI - Regulation of mitogen-activated protein kinases by sphingolipid products in oligodendrocytes. AB - Sphingolipid products such as ceramide (cer), sphingosine (sph), and sphingosine 1-phosphate (SPP) are implicated in the regulation of cell growth and apoptosis. We have recently shown that cer, sph, and SPP differentially modulate ionic events in cultured oligodendrocytes (OLGs). Cer but not sph or SPP inhibits the inward rectifier (I(Kir)) in OLGs. To further investigate the role of sphingolipid products in OLGs, we studied the effect of cer, sph, and SPP on OLG survival and on the regulation of mitogen-activated protein kinases (MAPKs). We found that cer, sph, and SPP differentially modulate OLG survival and activation of MAPK members. Cer causes OLG apoptosis, sph causes OLG lysis, and SPP does not affect OLG survival. Cer induces a preferential activation of p38alpha, whereas sph and SPP induce a preferential activation of extracellular signal-regulated kinase 2 (ERK2) in OLGs. In addition, the effect of cer on p38alpha activity is mimicked by the inhibition of I(Kir) with Ba(2+). In contrast, exposure to cer results in increased activity of ERK2 but not of p38alpha in astrocytes. Cer induced OLG apoptosis is attenuated by a p38 inhibitor, SB203580, and by expression of a p38alpha dominant negative mutant. We conclude that p38alpha is the mediator in cer-induced OLG apoptosis and that cer-induced I(Kir) inhibition may contribute to the sustained activation of p38alpha in OLGs. PMID- 10460253 TI - Cloning and expression of a queen pheromone-binding protein in the honeybee: an olfactory-specific, developmentally regulated protein. AB - Odorant-binding proteins (OBPs) are small abundant extracellular proteins thought to participate in perireceptor events of odor-pheromone detection by carrying, deactivating, and/or selecting odor stimuli. The honeybee queen pheromone is known to play a crucial role in colony organization, in addition to drone sex attraction. We identified, for the first time in a social insect, a binding protein called antennal-specific protein 1 (ASP1), which binds at least one of the major queen pheromone components. ASP1 was characterized by cDNA cloning, expression in Pichia pastoris, and pheromone binding. In situ hybridization showed that it is specifically expressed in the auxiliary cell layer of the antennal olfactory sensilla. The ASP1 sequence revealed it as a divergent member of the insect OBP family. The recombinant protein presented the exact characteristics of the native protein, as shown by mass spectrometry, and N terminal sequencing and exclusion-diffusion chromatography showed that recombinant ASP1 is dimeric. ASP1 interacts with queen pheromone major components, opposite to another putative honeybee OBP, called ASP2. ASP1 biosynthetic accumulation, followed by nondenaturing electrophoresis during development, starts at day 1 before emergence, in concomitance with the functional maturation of olfactory neurons. The isobar ASP1b isoform appears simultaneously to ASP1a in workers, but only at approximately 2 weeks after emergence in drones. Comparison of in vivo and heterologous expressions suggests that the difference between ASP1 isoforms might be because of dimerization, which might play a physiological role in relation with mate attraction. PMID- 10460254 TI - BAX translocation is a critical event in neuronal apoptosis: regulation by neuroprotectants, BCL-2, and caspases. AB - Members of the BCL-2 family of proteins either promote or repress programmed cell death. Here we report that neonatal sympathetic neurons undergoing apoptosis after nerve growth factor (NGF) deprivation exhibited a protein synthesis dependent, caspase-independent subcellular redistribution of BAX from cytosol to mitochondria, followed by a loss of mitochondrial cytochrome c and cell death. Treatment with elevated concentrations of the neuroprotectants KCl or cAMP at the time of deprivation prevented BAX translocation and cytochrome c release. However, administration of KCl or cAMP 12 hr after NGF withdrawal acutely prevented loss of mitochondrial cytochrome c, but not redistribution of BAX; rescue with NGF acutely prevented both events. Overexpression of Bcl-2 neither altered the normal subcellular localization of BAX nor prevented its redistribution with deprivation but did inhibit the subsequent release of cytochrome c, caspase activation, and cell death. Bcl-2 overexpression did not prevent cell death induced by cytoplasmic microinjection of cytochrome c into NGF deprived competent-to-die neurons. These observations suggest that the subcellular redistribution of BAX is a critical event in neuronal apoptosis induced by trophic factor deprivation. BCL-2 acts primarily, if not exclusively, at the level of mitochondria to prevent BAX-mediated cytochrome c release, whereas NGF, KCl, or cAMP may abort the apoptotic program at multiple checkpoints. PMID- 10460255 TI - cAMP-dependent protein kinase phosphorylations on tau in Alzheimer's disease. AB - To elucidate the role cAMP-dependent protein kinase (PKA) phosphorylations on tau play in Alzheimer's disease, we have generated highly specific monoclonal antibodies, CP-3 and PG-5, which recognize the PKA-dependent phosphorylations of ser214 and ser409 in tau respectively. The present study demonstrates by immunohistochemical analysis, CP-3 and PG-5 immunoreactivity with neurofibrillary pathology in both early and advanced Alzheimer's disease, but not in normal brain tissue and demonstrates that cAMP-dependent protein kinase phosphorylations on tau precede or are coincident with the initial appearance of filamentous aggregates of tau. Studies using heat-stable preparations demonstrate that neither site appears to be phosphorylated to any appreciable extent in normal rodent or human brain. Further analysis demonstrates that the beta catalytic subunit of PKA (Cbeta), the beta II regulatory subunit of PKA (RIIbeta), and the 79 kDa A-kinase-anchoring-protein (AKAP79), are tightly associated with the neurofibrillary pathology, positioning cAMP-dependent protein kinase to participate directly in the pathological hyperphosphorylation of tau seen in Alzheimer's disease. PMID- 10460256 TI - Mitochondrial clearance of cytosolic Ca(2+) in stimulated lizard motor nerve terminals proceeds without progressive elevation of mitochondrial matrix [Ca(2+)]. AB - This study used fluorescent indicator dyes to measure changes in cytosolic and mitochondrial [Ca(2+)] produced by physiological stimulation of lizard motor nerve terminals. During repetitive action potential discharge at 10-50 Hz, the increase in average cytosolic [Ca(2+)] reached plateau at levels that increased with increasing stimulus frequency. This stabilization of cytosolic [Ca(2+)] was caused mainly by mitochondrial Ca(2+) uptake, because drugs that depolarize mitochondria greatly increased the stimulation-induced elevation of cytosolic [Ca(2+)], whereas blockers of other Ca(2+) clearance routes had little effect. Surprisingly, during this sustained Ca(2+) uptake the free [Ca(2+)] in the mitochondrial matrix never exceeded a plateau level of approximately 1 microM, regardless of stimulation frequency or pattern. When stimulation ceased, matrix [Ca(2+)] decreased over a slow ( approximately 10 min) time course consisting of an initial plateau followed by a return to baseline. These measurements demonstrate that sustained mitochondrial Ca(2+) uptake is not invariably accompanied by progressive elevation of matrix free [Ca(2+)]. Both the plateau of matrix free [Ca(2+)] during stimulation and its complex decay after stimulation could be accounted for by a model incorporating reversible formation of an insoluble Ca salt. This mechanism allows mitochondria to sequester large amounts of Ca(2+) while maintaining matrix free [Ca(2+)] at levels sufficient to activate Ca(2+)-dependent mitochondrial dehydrogenases, but below levels that activate the permeability transition pore. PMID- 10460257 TI - Disabled-1 binds to the cytoplasmic domain of amyloid precursor-like protein 1. AB - Disruption of the disabled-1 gene (Dab1) results in aberrant migration of neurons during development and disorganization of laminar structures throughout the brain. Dab1 is thought to function as an adapter molecule in signal transduction processes. It contains a protein-interaction (PI) domain similar to the phosphotyrosine-binding domain of the Shc oncoprotein, it is phosphorylated by the Src protein tyrosine kinase, and it binds to SH2 domains in a phosphotyrosine dependent manner. To investigate the function of Dab1, we searched for binding proteins using the yeast two-hybrid system. We found that the PI domain of Dab1 interacts with the amyloid precursor-like protein 1 (APLP1). The association of Dab1 with APLP1 was confirmed in biochemical assays, and the site of interaction was localized to a cytoplasmic region of APLP1 containing the amino acid sequence motif Asn-Pro-x-Tyr (NPxY). NPxY motifs are involved in clathrin-mediated endocytosis, and they have been shown to bind to PI domains present in several proteins. This region of APLP1 is conserved among all members of the amyloid precursor family of proteins. Indeed, we found that Dab1 also interacts with amyloid precursor protein (APP) and APLP2 in biochemical association experiments. In transiently transfected cells, Dab1 and APLP1 colocalized in membrane ruffles and vesicular structures. Cotransfection assays in cultured cells indicated that APP family members increased serine phosphorylation of Dab1. Dab1 and APLP1 are expressed in similar cell populations in developing and adult brain tissue. These results suggest that Dab1 may function, at least in part, through association with APLP1 in the brain. PMID- 10460258 TI - Dependence of nodal sodium channel clustering on paranodal axoglial contact in the developing CNS. AB - Na(+) channel clustering at nodes of Ranvier in the developing rat optic nerve was analyzed to determine mechanisms of localization, including the possible requirement for glial contact in vivo. Immunofluorescence labeling for myelin associated glycoprotein and for the protein Caspr, a component of axoglial junctions, indicated that oligodendrocytes were present, and paranodal structures formed, as early as postnatal day 7 (P7). However, the first Na(+) channel clusters were not seen until P9. Most of these were broad, and all were excluded from paranodal regions of axoglial contact. The number of detected Na(+) channel clusters increased rapidly from P12 to P22. During this same period, conduction velocity increased sharply, and Na(+) channel clusters became much more focal. To test further whether oligodendrocyte contact directly influences Na(+) channel distributions, nodes of Ranvier in the hypomyelinating mouse Shiverer were examined. This mutant has oligodendrocyte-ensheathed axons but lacks compact myelin and normal axoglial junctions. During development Na(+) channel clusters in Shiverer mice were reduced in numbers and were in aberrant locations. The subcellular location of Caspr was disrupted, and nerve conduction properties remained immature. These results indicate that in vivo, Na(+) channel clustering at nodes depends not only on the presence of oligodendrocytes but also on specific axoglial contact at paranodal junctions. In rats, ankyrin-3/G, a cytoskeletal protein implicated in Na(+) channel clustering, was detected before Na(+) channel immunoreactivity but extended into paranodes in non-nodal distributions. In Shiverer, ankyrin-3/G labeling was abnormal, suggesting that its localization also depends on axoglial contact. PMID- 10460259 TI - Polysialylated neural cell adhesion molecule-positive CNS precursors generate both oligodendrocytes and Schwann cells to remyelinate the CNS after transplantation. AB - Transplantation offers a means of identifying the differentiation and myelination potential of early neural precursors, features relevant to myelin regeneration in demyelinating diseases. In the postnatal rat brain, precursor cells expressing the polysialylated (PSA) form of the neural cell adhesion molecule NCAM have been shown to generate mostly oligodendrocytes and astrocytes in vitro (Ben-Hur et al., 1998). Immunoselected PSA-NCAM+ newborn rat CNS precursors were expanded as clusters with FGF2 and grafted into a focal demyelinating lesion in adult rat spinal cord. We show that these neural precursors can completely remyelinate such CNS lesions. While PSA-NCAM+ precursor clusters contain rare P75+ putative neural crest precursors, they do not generate Schwann cells in vitro even in the presence of glial growth factor. Yet they generate oligodendrocytes, astrocytes, and Schwann cells in vivo when confronted with demyelinated axons in a glia-free area. We confirmed the transplant origin of these Schwann cells using Y chromosome in situ hybridization and immunostaining for the peripheral myelin protein P0 of tissue from female rats that had been grafted with male cell clusters. The number and distribution of Schwann cells within remyelinated tissue, and the absence of P0 mRNAs in donor cells, indicated that Schwann cells were generated by expansion and differentiation of transplanted PSA-NCAM+ neural precursors and were not derived from contaminating Schwann cells. Thus, transplantation into demyelinated CNS tissue reveals an unexpected differentiation potential of a neural precursor, resulting in remyelination of CNS axons by PNS and CNS myelin-forming cells. PMID- 10460260 TI - Inactivation of Rho signaling pathway promotes CNS axon regeneration. AB - Regeneration in the CNS is blocked by many different growth inhibitory proteins. To foster regeneration, we have investigated a strategy to block the neuronal response to growth inhibitory signals. Here, we report that injured axons regrow directly on complex inhibitory substrates when Rho GTPase is inactivated. Treatment of PC12 cells with C3 enzyme to inactivate Rho and transfection with dominant negative Rho allowed neurite growth on inhibitory substrates. Primary retinal neurons treated with C3 extended neurites on myelin-associated glycoprotein and myelin substrates. To explore regeneration in vivo, we crushed optic nerves of adult rat. After C3 treatment, numerous cut axons traversed the lesion to regrow in the distal white matter of the optic nerve. These results indicate that targeting signaling mechanisms converging to Rho stimulates axon regeneration on inhibitory CNS substrates. PMID- 10460261 TI - Neuropil pattern formation and regulation of cell adhesion molecules in Drosophila optic lobe development depend on synaptobrevin. AB - To investigate a possible involvement of synaptic machinery in Drosophila visual system development, we studied the effects of a loss of function of neuronal synaptobrevin, a protein required for synaptic vesicle release. Expression of tetanus toxin light chain (which cleaves neuronal synaptobrevin) and genetic mosaics were used to analyze neuropil pattern formation and levels of selected neural adhesion molecules in the optic lobe. We show that targeted toxin expression in the developing optic lobe results in disturbances of the columnar organization of visual neuropils and of photoreceptor terminal morphology. IrreC rst immunoreactivity in neuropils is increased after widespread expression of toxin. In photoreceptors, targeted toxin expression results in increased Fasciclin II and chaoptin but not IrreC-rst immunoreactivity. Axonal pathfinding and programmed cell death are not affected. In genetic mosaics, patches of photoreceptors that lack neuronal synaptobrevin exhibit the same phenotypes observed after photoreceptor-specific toxin expression. Our results demonstrate the requirement of neuronal synaptobrevin for regulation of cell adhesion molecules and development of the fine structure of the optic lobe. A possible causal link to fine-tuning processes that may include synaptic plasticity in the development of the Drosophila CNS is discussed. PMID- 10460262 TI - Identification of an interneuronal population that mediates recurrent inhibition of motoneurons in the developing chick spinal cord. AB - Studies on the development of synaptic specificity, embryonic activity, and neuronal specification in the spinal cord have all been limited by the absence of a functionally identified interneuron class (defined by its unique set of connections). Here, we identify an interneuron population in the embryonic chick spinal cord that appears to be the avian equivalent of the mammalian Renshaw cell (R-interneurons). These cells receive monosynaptic nicotinic, cholinergic input from motoneuron recurrent collaterals. They make predominately GABAergic connections back onto motoneurons and to other R-interneurons but project rarely to other spinal interneurons. The similarity between the connections of the developing R-interneuron, shortly after circuit formation, and the mature mammalian Renshaw cell raises the possibility that R-interneuronal connections are formed precisely from the onset. Using a newly developed optical approach, we identified the location of R-interneurons in a column, dorsomedial to the motor nucleus. Functional characterization of the R-interneuron population provides the basis for analyses that have so far only been possible for motoneurons. PMID- 10460263 TI - Mechanisms of induction and expression of long-term depression at GABAergic synapses in the neonatal rat hippocampus. AB - Synaptic plasticity at excitatory glutamatergic synapses is believed to be instrumental in the maturation of neuronal networks. Using whole-cell patch-clamp recordings, we have studied the mechanisms of induction and expression of long term depression at excitatory GABAergic synapses in the neonatal rat hippocampus (LTD(GABA-A)). We report that the induction of LTD(GABA-A) requires a GABA(A) receptor-mediated membrane depolarization, which is necessary to remove the Mg(2+) block from postsynaptic NMDA receptors. LTD(GABA-A) is associated with an increase in the coefficient of variation of evoked GABA(A) receptor-mediated synaptic currents and a decrease in the frequency, but not amplitude, of Sr(2+) induced asynchronous GABA(A) quantal events. We conclude that LTD(GABA-A) induction requires the activation of both GABA(A) and NMDA postsynaptic receptors and that its expression is likely presynaptic. PMID- 10460264 TI - Evidence for brainstem and supra-brainstem contributions to rapid cortical plasticity in adult monkeys. AB - Cortical maps can undergo amazingly rapid changes after injury of the body. These changes involve functional alterations in normal substrates, but the cortical and/or subcortical location(s) of these alterations, and the relationships of alterations in different substrates, remain controversial. The present study used neurophysiological approaches in adult monkeys to evaluate how brainstem organization of tactile inputs in the cuneate nucleus (CN) changes after acute injury of hand nerves. These data were then compared with analogous data from our earlier cortical area 3b studies, which used the same approaches and acute injury, to assess relationships of cuneate and cortical changes. The results indicate that cuneate tactile responsiveness, receptive field locations, somatotopic organization, and spatial properties of representations (i.e., location, continuity, size) change during the first minutes to hours after injury. The comparisons of cuneate and area 3b organization further show that some cuneate changes are preserved in area 3b, whereas other cuneate changes are transformed before being expressed in area 3b. The findings provide evidence that rapid reorganization in area 3b, in part, reflects mechanisms that operate from a distance in the cuneate nucleus and, in part, reflects supracuneate mechanisms that modify brainstem changes. PMID- 10460265 TI - Effects of attention on the processing of motion in macaque middle temporal and medial superior temporal visual cortical areas. AB - The visual system is continually inundated with information received by the eyes. Only a fraction of this information appears to reach visual awareness. This process of selection is one of the functions ascribed to visual attention. Although many studies have investigated the role of attention in shaping neuronal representations in cortical areas, few have focused on attentional modulation of neuronal signals related to visual motion. We recorded from 89 direction selective neurons in middle temporal (MT) and medial superior temporal (MST) visual cortical areas of two macaque monkeys using identical sensory stimulation under various attentional conditions. Neural responses in both areas were greatly influenced by attention. When attention was directed to a stimulus inside the receptive field of a neuron, responses in MT and MST were enhanced an average of 20 and 40% compared with a condition in which attention was directed outside the receptive field. Even stronger average enhancements (70% in MT and 100% in MST) were observed when attention was switched from a stimulus moving in the nonpreferred direction inside the receptive field to another stimulus in the receptive field that was moving in the preferred direction. These findings show that attention modulates motion processing from stages early in the dorsal visual pathway by selectively enhancing the representation of attended stimuli and simultaneously reducing the influence of unattended stimuli. PMID- 10460266 TI - Behavioral modulation of tactile responses in the rat somatosensory system. AB - We investigated the influence of four different behavioral states on tactile responses recorded simultaneously via arrays of microwires chronically implanted in the vibrissal representations of the rat ventral posterior medial nucleus (VPM) of the thalamus and the primary somatosensory cortex (SI). Brief (100 microsecond) electrical stimuli delivered via a cuff electrode to the infraorbital nerve yielded robust sensory responses in VPM and SI during states of quiet immobility. However, significant reductions in tactile response magnitude and latency were observed in VPM and SI during large-amplitude, exploratory movements of the whiskers (at approximately 4-6 Hz). During small amplitude, 7-12 Hz whisker-twitching movements, a significant reduction in SI response magnitude and an increase in VPM and SI response latencies were observed as well. When pairs of stimuli with interstimulus intervals <100 msec were delivered during quiet immobility, the response to the second stimulus in the pair was reduced and occurred at a longer latency compared with the response to the first stimulus. In contrast, during large-amplitude whisker movements and general motor activity, paired stimuli yielded similar sensory responses at interstimulus intervals >25 msec. These response patterns were correlated with the amount and duration of postexcitatory firing suppression observed in VPM and SI during each of these behaviors. On the basis of these results, we propose that sensory responses are dynamically modulated during active tactile exploration to optimize detection of different types of stimuli. During quiet immobility, the somatosensory system seems to be optimally tuned to detect the presence of single stimuli. In contrast, during whisker movements and other exploratory behaviors, the system is primed to detect the occurrence of rapid sequences of tactile stimuli, which are likely to be generated by multiple whisker contacts with objects during exploratory activity. PMID- 10460267 TI - Mechanisms underlying spontaneous oscillation and rhythmic firing in rat subthalamic neurons. AB - Subthalamic neurons drive basal ganglia output neurons in resting animals and relay cortical and thalamic activity to the same output neurons during movement. The first objective of this study was to determine the mechanisms underlying the spontaneous activity of subthalamic neurons in vitro and to gain insight into their resting discharge in vivo. The second objective was to determine the response of subthalamic neurons to depolarizing current injection and how intrinsic properties may shape their response to cortical and thalamic inputs during movement. Cell-attached and whole-cell recordings were made from subthalamic neurons in brain slices prepared from 3- to 4-week-old rats. The slow, rhythmic discharge of subthalamic neurons was resistant to blockade of excitatory synaptic transmission indicating that intrinsic currents underlie their spontaneous discharge. A persistent sodium current was the source of current during the depolarizing phase of the oscillation. A powerful afterhyperpolarization following each action potential was sufficient to terminate the depolarization. A long duration component of the spike afterhyperpolarization determined the period of the oscillation and was generated by an apamin-sensitive calcium-activated potassium current. Calcium entry responsible for that current was associated with action potentials. Subthalamic neurons exhibited a sigmoidal frequency-current relationship with the steeper portion starting at approximately 30-40 Hz. This property makes subthalamic neurons more sensitive to input at high firing rates associated with movement than at low rates associated with rest. We propose that the subthreshold persistent sodium current overcomes calcium activated potassium current which accumulates during high frequency firing and underlies the enhanced sensitivity to current >30 Hz. PMID- 10460268 TI - Frequency-dependent PSP depression contributes to low-pass temporal filtering in Eigenmannia. AB - This study examined the contribution of frequency-dependent short-term depression of PSP amplitude to low-pass temporal filtering in the weakly electric fish Eigenmannia. Behavioral and neurophysiological methods were used. Decelerations of the electric organ discharge frequency were measured in response to continuous and discontinuous electrosensory stimuli. Decelerations were strongest (median = 4.7 Hz; range, 3.5-5.9 Hz) at continuous beat rates of approximately 5 Hz and weakest (median = 0.4 Hz; range, 0.0-0.8 Hz) at beat rates of 30 Hz. Gating 20 or 30 Hz stimuli at a rate of 5 Hz, however, elicited decelerations that were sixfold greater than that of continuous stimuli at these beat rates (median = 2.6 Hz; range, 2.0-4.7 Hz for 30 Hz). These results support the hypothesis that short term processes enhance low-pass filtering by reducing responses to fast beat rates. This hypothesis was tested by recording intracellularly the responses of 33 midbrain neurons to continuous and discontinuous stimuli. Results indicate that short-term depression of PSP amplitude primarily accounts for the steady state low-pass filtering of these neurons beyond that contributed by their passive and active membrane properties. Previous results demonstrate that passive properties can contribute up to 7 dB of low-pass filtering; PSP depression can add up to an additional 12.5 dB (median = 4.5). PSP depression increased in magnitude with stimulus frequency and showed a prominent short-term component (t(1) = 66 msec at 30 Hz). Initial PSP amplitude recovered fully after a gap of 150 msec for most neurons. Remarkably, recovery of PSP amplitude could be produced by inserting a brief low-temporal frequency component in the stimulus. PMID- 10460269 TI - Nitric oxide stimulates ACTH secretion and the transcription of the genes encoding for NGFI-B, corticotropin-releasing factor, corticotropin-releasing factor receptor type 1, and vasopressin in the hypothalamus of the intact rat. AB - We investigated the effect of the intracerebroventricular injection of the nitric oxide (NO) donor 3-morpholino-sydnonimine (SIN-1) on the release of adrenocorticotropin hormone (ACTH) and the neuronal response of hypothalamic neurons responsible for this release. Rats that were administered SIN-1 showed significant elevations in plasma ACTH levels, a response that was virtually abolished by antibodies against corticotropin-releasing factor (CRF) and significantly blunted by vasopressin (VP) antiserum. SIN-1 also upregulated heteronuclear (hn) transcripts for CRF and VP and messenger RNA (mRNA) levels for the immediate early gene NGFI-B and for CRF receptor type 1 (CRF-R(1)) in the parvocellular portion of the paraventricular nucleus (PVN) of the hypothalamus. Blockade of prostaglandin synthesis with ibuprofen did not alter the ACTH or the PVN response to SIN-1. The central nucleus of the amygdala and the supraoptic nucleus, regions that are involved in autonomic adjustments to altered cardiovascular activity, also responded to SIN-1 with elevated NGFI-B mRNA levels. However, the only change in mean arterial blood pressure caused by this NO donor was a transient and modest increase. To our knowledge, this is the first demonstration that in the intact rat NO stimulates the activity of PVN neurons that control the hypothalamic-pituitary-adrenal axis. It must be noted, however, that our results do not allow us to determine whether this effect was direct or mediated through PVN afferents. This study should help resolve the controversy generated by the use of isolated brain tissues to investigate the net effect of NO on hypothalamic peptide production. PMID- 10460270 TI - Lateral hypothalamic serotonin inhibits nucleus accumbens dopamine: implications for sexual satiety. AB - Dopamine (DA) is released in several brain areas, including the nucleus accumbens (NAcc), before and during copulation in male rats. DA agonists administered into this area facilitate, and DA antagonists inhibit, numerous motivated behaviors, including male sexual behavior. Serotonin (5-HT) is generally inhibitory to male sexual behavior. We reported previously that 5-HT is released in the anterior lateral hypothalamic area (LHA(A)) and that a selective serotonin reuptake inhibitor microinjected into that area delayed and slowed copulation. Our present results, using high temporal resolution microdialysis, (1) confirm previous electrochemical evidence that extracellular levels of DA increase in the NAcc during copulation and decrease during the postejaculatory interval (PEI) and (2) reveal that LHA(A) 5-HT can inhibit both basal and female-elicited DA release in the NAcc. These findings suggest that the neural circuit promoting sexual quiescence during the PEI includes serotonergic input to the LHA(A), which in turn inhibits DA release in the NAcc. These findings may also provide insights concerning the inhibitory control of other motivated behaviors activated by the NAcc and may have relevance for understanding the sexual side effects common to antidepressant medications. PMID- 10460271 TI - Recovery of presynaptic dopaminergic functioning in rats treated with neurotoxic doses of methamphetamine. AB - Repeated administration of methamphetamine (METH) to animals can result in long lasting decreases in striatal dopamine (DA) content. In addition, the evoked overflow of striatal DA is reduced in rats 1 week after neurotoxic doses of METH. However, whether these functional changes in DA release are permanent or tend to recover over time has not been established. In the present study we used in vivo electrochemistry and microdialysis to examine evoked overflow of DA in the striatum of METH-treated rats at several time points after treatment to determine if DA overflow would spontaneously recover. Male Fischer-344 rats were administered METH (5 mg/kg, s.c. ) or saline four times in one day at 2 hr intervals. In vivo electrochemistry experiments in anesthetized rats, and in vivo microdialysis studies in awake rats, were carried out 1 week, 1 month, 6 months, and 12 months after treatment. At 1 week after treatment there were significant decreases in potassium- and amphetamine-evoked overflow of DA, and in clearance of DA, in the striatum of the METH-treated animals. Basal extracellular levels of DA and its metabolites were also decreased. Evoked overflow had partially recovered by 1 month. By 6 months evoked overflow of DA appeared to be normal in the METH-treated rats. However, whole tissue levels of striatal DA were still significantly decreased. All parameters were back to control values by 12 months. These results suggest that presynaptic dopaminergic functioning can recover to normal levels in the striatum of METH-treated rats by 12 months after treatment. PMID- 10460272 TI - Redundant basal forebrain modulation in taste aversion memory formation. AB - Mnemonic deficits resulting from excitotoxic lesion of the basal forebrain have been classically attributed to the resulting depletion of cortical acetylcholine activity. It has been demonstrated that in spite of the strong cholinergic depletion after injections into the basal forebrain of the immunotoxin 192IgG saporin, no detectable deficit can be found in the acquisition of several learning tasks, including conditioned taste aversion. Conversely, NMDA-induced lesions of the basal forebrain strongly impair taste aversion learning. In this study we show that 192IgG-saporin produces an efficient and selective cholinergic deafferentation of the rat neocortex but not the amygdala. Furthermore, a stronger relationship between severity of memory impairment after NMDA lesions and basoamygdaloid cholinergic deafferentation was found. Therefore, in a second experiment, we show that combining NMDA-induced lesions into the basolateral amygdala with 192IgG-saporin injections into the basal forebrain results in a strong disruption of taste aversion learning, whereas none of these treatments were by themselves capable of producing any detectable impairment in this learning task. The double lesion effect was only paralleled by simple NMDA lesions into the basal forebrain, suggesting that the learning deficits associated to excitotoxic lesions of the basal forebrain are the result of the simultaneous destruction of the corticopetal and basoamygdaloid interaction. A model is proposed, according to which the modulation of learning processes exerted by the basal forebrain can be redundantly performed by both the basocortical and basoamygdaloid pathway. PMID- 10460273 TI - Spinal substance P receptor expression and internalization in acute, short-term, and long-term inflammatory pain states. AB - Inflammatory pain involves the sensitization of both primary afferent and spinal cord neurons. To explore the neurochemical changes that contribute to inflammatory pain, we have examined the expression and ligand-induced internalization of the substance P receptor (SPR) in the spinal cord in acute, short-term, and long-term inflammatory pain states. These inflammatory models included unilateral injection of formalin (8-60 min), carrageenan (3 hr), and complete Freund's adjuvant (CFA; 3 d) into the rat hindpaw as well as adjuvant induced polyarthritis (21 d). In acute inflammatory pain there is ongoing release of substance P (SP) as measured by SPR internalization in lamina I neurons at both 8 and 60 min after formalin injection. Although there is no tonic release of SP in short-term inflammatory pain, at 3 hr after carrageenan injection, SP is released in response to both noxious and non-noxious somatosensory stimulation with SPR internalization being observed in neurons located in both laminae I and III-IV. In long-term inflammatory pain models (CFA and polyarthritis) the same pattern of SP release and SPR activation occurs as is observed in short-term inflammation with the addition that there is a significant upregulation of the SPR in lamina I neurons. These results suggest that SPR internalization might serve as a marker of the contribution of ongoing primary afferent input in acute and persistent pain states. These stereotypical neurochemical changes suggest that there are unique neurochemical signatures for acute, short-term, and long term inflammatory pain. PMID- 10460274 TI - Reorganization in primary motor cortex of primates with long-standing therapeutic amputations. AB - Intracortical microstimulation was used to investigate the organization of primate primary motor cortex (M1) in three squirrel monkeys and two galagos years after the therapeutic amputation of an injured forelimb or hindlimb. In two squirrel monkeys with forelimb amputation, physiological results were correlated with the distribution of corticospinal neurons after injections of tracers into the lower cervical segments of the spinal cord. Distributions of labeled corticospinal neurons helped identify the locations of the former forelimb cortex in M1. Evoked movements from M1 ipsilateral to the missing limb were not obviously different from M1 of normal controls. Stimulation in the deefferented part of M1 contralateral to the missing limb elicited movements of the remaining proximal muscles as well as movements from adjacent body representations in all cases. Stimulation in the deefferented forelimb cortex evoked shoulder stump, trunk, and orofacial movements, whereas stimulation in the deefferented hindlimb cortex evoked hip stump, trunk, and tail movements. Movements were evoked from all sites in the deprived cortex, so that there were no unresponsive zones. Minimal levels of current necessary to evoke these movements varied from those in the normal range to those of much higher levels, with the average threshold higher than normal. Finally, multiunit recording from the two galagos revealed that the deprived portions of S1 were responsive to touch or taps on the stump and neighboring body parts. PMID- 10460275 TI - Dopaminergic modulation of voltage-gated Na+ current in rat hippocampal neurons requires anchoring of cAMP-dependent protein kinase. AB - Activation of D1-like dopamine (DA) receptors reduces peak Na(+) current in acutely isolated hippocampal neurons via a modulatory mechanism involving phosphorylation of the Na(+) channel alpha subunit by cAMP-dependent protein kinase (PKA). Peak Na(+) current is reduced 20-50% in the presence of the D1 agonist SKF 81297 or the PKA activator Sp-5,6-dichloro-l-beta-d-ribofuranosyl benzimidazole-3',5'-cyclic monophosphorothionate (cBIMPS). Co-immunoprecipitation experiments show that Na(+) channels are associated with PKA and A-kinase anchoring protein 15 (AKAP-15), and immunocytochemical labeling reveals their co localization in the cell bodies and proximal dendrites of hippocampal pyramidal neurons. Anchoring of PKA near the channel by an AKAP, which binds the RII alpha regulatory subunit, is necessary for Na(+) channel modulation in acutely dissociated hippocampal pyramidal neurons. Intracellular dialysis with the anchoring inhibitor peptides Ht31 from a human thyroid AKAP and AP2 from AKAP-15 eliminated the modulation of the Na(+) channel by the D1-agonist SKF 81297 and the PKA activator cBIMPS. In contrast, dialysis with the inactive proline substituted control peptides Ht31-P and AP2-P had little effect on the D1 and PKA modulation. Therefore, we conclude that modulation of the Na(+) channel by activation of D1-like DA receptors requires targeted localization of PKA near the channel to achieve phosphorylation of the alpha subunit and to modify the functional properties of the channel. PMID- 10460277 TI - [Biomaterials in the human]. PMID- 10460276 TI - Growth-associated protein 43 is located in type I corticothalamic terminals in the cat visual thalamus. AB - Growth-associated protein 43 (GAP 43) is a presynaptic protein that has been proposed to be involved in synaptic plasticity. To determine the location of GAP 43 within the synaptic circuitry of the thalamus, immunocytochemical staining for GAP 43 was examined in a relay nucleus, the dorsal lateral geniculate nucleus (dLGN), and two association nuclei, the pulvinar nucleus and the lateral subdivision of the lateral posterior (LP) nucleus. In the dLGN, moderate neuropil staining was seen in the A laminae, and denser staining was found in the interlaminar zones and the C laminae. Uniform dense staining of the neuropil was found in both the pulvinar and LP nuclei. At the ultrastructural level, the GAP 43 staining was restricted to small-diameter myelinated axons, thin unmyelinated fibers, and small terminals that contained densely packed round vesicles (RS profiles) and made asymmetric synaptic contacts with small-caliber dendrites in the extraglomerular neuropil. The distribution of immunocytochemical label within the visual thalamus suggests that GAP 43 is confined to type I corticothalamic terminals and axons that originate from extrastriate cortical areas. These results also suggest that in both relay and association nuclei GAP 43 may be used to augment the cortical control of thalamic activity. In addition, these results underscore the distinction between the small type I corticothalamic terminals, which appear to contain GAP 43 throughout the visual thalamus, and the large type II corticothalamic terminals that, like the type II retinal terminals in the dLGN, do not contain GAP 43. PMID- 10460278 TI - [Biomaterials, human tolerance and integration]. AB - Biomaterials and related process engineering in order to obtain optimal surface and structural biocompatibility of implants and devices are presented. Vital avital composites for tissue engineering, cell culture models, porous ceramics and degradable polymers are introduced as examples. Emphasis is laid on the conversion of basic research results into clinical applications and on the exchange of technologies from the non-medical into the medical field and vice versa. PMID- 10460279 TI - [Endovascular prostheses: evolution, state-of-the-art and perspectives of a new technology]. AB - The treatment of infrarenal aortic aneurysms by means of transluminally placed endovascular prostheses reflects significant progress in the field of vascular surgery. In the case of infrarenal aortic aneurysm it is possible to achieve technically successful implantation of such a prosthesis in well over 90 % of cases. The rate of clinical success, meaning lasting effective exclusion of the aortic aneurysm, cannot (yet) be definitively determined, since no long-term results are so far available. Secondary leaks are observed in at least 10 % of all patients, making a further therapy necessary (endorepair, conversion, embolization). Further development of endovascular prostheses will include optimization of the aortal/iliac attachment of the prostheses, a better configuration and the development of long-lasting materials that can be used for endovascular prostheses. PMID- 10460280 TI - [Metal stents in gastroenterology]. AB - Self-expanding metal stents have, when introduced in their constrained form through gastrointestinal and biliary strictures, a relatively small diameter. Once placed through the stricture and released, however, they expand to a much larger internal diameter, thus giving rise to sufficient palliative reopening of these strictures. Since metal stents are usually not removable, their primary use has been in malignant stenoses. Benign strictures should be treated with metal stents only in exceptional situations. For palliation of dysphagia in esophagocardial malignancies, metal stents have been shown to be associated with significantly fewer initial complications on placement than plastic tubes. The long-term fate of both stent types seems to be similar. In the palliation of malignant jaundice, metal stents were demonstrated to have a significantly longer patency rate, resulting in a favourable cost-benefit ratio despite the high price of metal stents. Continued developments are necessary to further reduce long-term complications and effectiveness. PMID- 10460281 TI - [Meshes within the abdominal wall]. AB - The marked reduction in recurrence rates following reinforcement of the abdominal wall by meshes in incisional hernia has promoted their increasingly widespread use. The primary suture in technique failed more than half of the cases; therefore, the closure method needs to be changed and improved, particularly with regard to a possibly underlying defect in collagen metabolism. After more than 100 years of mesh development they are mainly placed in a sublay or onlay position, ePTFE, polyester and polypropylene are preferred. In any case the mesh has to overlap the defect sufficiently because of wound contraction. On the basis of our experience and reports in the literature, the advantages and disadvantages of various mesh techniques and mesh materials are discussed. However, because long-term studies are missing, the relevance of the cumulative risk for long-term complications such as mesh migration and fistula formation, the extent of patient complaints or the potential risk of a persistent foreign-body reaction cannot yet be ascertained. Nevertheless, because there are no surgical alternatives, meshes represent an improvement in hernia surgery that cannot be overestimated. PMID- 10460282 TI - [Lebertransplantation mit Erhalt der V. cava und temporarem portocavalem Shunt] PMID- 10460283 TI - [Biomaterials in the skeletal system]. AB - The increasing number of biomaterials for the skeletal system requires mote and move their distinct clinical application. To guarantee useful therapeutic results the characteristics of the biomaterials have to be matched with the characteristics of the implantation site. Recently developed biodegradable polymers with a slow degradation process and longer stability are being increasingly clinically applied with a low complication rate. The development of new remodelable bone cements for bone-defect filling revives the idea of cement metallic implant constructs. Finally, recombinant human bone growth factors are currently under controlled clinical examination with first promising results. Long-term results allowing common clinical use of these factors are still to be expected. PMID- 10460284 TI - [Immune regulation of wound healing]. AB - Wound healing is a complex cascade of biochemical and cellular events designed to achieve restoration of tissue integrity following injury. Immune cells are critical for the outcome of healing. Much has been learned about the individual events that are involved in this process. Knowledge is scant, however, about the spatial and temporal interweaving of these events and how one step sets the stage for subsequent observed phenomena. Imbalance of stimulating and inhibiting factors causes failure of healing. Immunosuppressive states interfere with a normal immune response and impair wound repair. Clinically, new therapeutic approaches to modulate impaired healing are based on a knowledge of immune regulation of wound healing. PMID- 10460286 TI - [The influence of different therapeutic approaches on the survival of elderly burn patients]. AB - The present study analyzes the outcome of 51 patients aged more than 65 years, who were admitted between 1993 and 1997 to a specific burns unit with the possibility of intensive care medicine including respirator therapy and hemofiltration. The results were compared to an historical control group treated between 1980 and 1990. The overall survival rate was 54 %. Analyzing the patients' mortality, an increase in the number of nurses, improvement of enteral nutrition and wound monitoring had more influence on the survival rates than respirator therapy, hemofiltration and catecholamines. This is in contrast to our experience with younger individuals. The positive influence of intensive care on survival was restricted only to a small number of cases. The group of patients who died on the intensive care unit showed a relatively long median survival time of 24 days. Septic multiorgan failure was the main cause of death. PMID- 10460285 TI - [Results of lung-volume reduction surgery in end-stage lung emphysema]. AB - BACKGROUND: Surgical treatment of diffuse, nonbullous emphysema was first described by Brantigan et al. in 1957 and was reintroduced by Cooper et al. in 1995 as lung-volume reduction surgery (LVRS). Meanwhile it has become an internationally established procedure in the treatment of lung emphysema. We report our results after LVRS in 91 patients. METHODS: Between September 1994 and August 1998 LVRS was performed through median sternotomy (n = 15), videoendoscopy (n = 49), thoracotomy (n = 18) or combined video endoscopy on one side and thoracotomy on the other (n = 9) in 91 patients (aged 33-80 years; mean 56,4 years). All patients showed progression of severe dyspnea despite maximum medical and physical therapy. RESULTS: Perioperative mortality was 5.5 % (5 patients). Seventy-five percent of the patients showed significant functional improvement [postoperatively above 120 % of the preoperative forced expiratory volume in 1 s (FeV1)]. Mean FeV1 significantly increased by 28.6 % from 25.5 +/- 1.2 % predicted (% p) preoperatively to 32.8 +/- 1.9 % p during the first 6 months postoperatively (p < 0.005). Furthermore, the mean RV was reduced from 320.3 +/- 7.9 % p preoperatively to 248.4 +/- 7.5 % p 6 months postoperatively and mean TLC from 140.2 +/- 2.4 % p to 126.1 +/- 2.1 % p (p < 0.005). Intrinsic PEEP decreased significantly from 5.1 +/- 0.4 cm H(2)O preoperatively to 2.3 +/- 0.3 cm H(2)O postoperatively (p < 0.05). CONCLUSION: LVRS is an excellent therapeutic option for selected patients with severe emphysema and additional signs of severe hyperinflation. It results in significant postoperative functional improvement and marked increase in quality of life. The long-term benefit of LVRS remains to be defined. PMID- 10460287 TI - [Kommentar auf Anforderung der Schriftleitung] PMID- 10460288 TI - [Learning and optimising minimally invasive techniques by operating on embalmed human corpses]. AB - The expansion of minimally invasive surgery worldwide provokes an intensive interest of all surgical disciplines in gaining possibilities for research, learning and teaching by operating on human corpses. Despite the fact that German anatomical institutes in general have the infrastructure to realise such clinical cooperations, at present they may offer such opportunities only to a restricted degree, since the concentration on student teaching and anatomic research limits the capacity of the staff for a commitment in this field of applied science. To provide a basis for future estimations of the efforts necessary to perform solo surgery on the human cadaver, especially with emphasis on research, quality control and teaching, this article reports on practical experiences with such a project named "ANAtoMIC", identifying minimal conditions which have to be realised. PMID- 10460289 TI - [Recanalization of extensive arterial occlusion in the thigh using an endovascular PTFE stent graft (Gore Hemobahn) as an alternative to a femoropopliteal P1 bypass]. AB - A few months ago an endovascular implantable PTFE stent graft became commercially available. Since May 1998 we have treated 15 patients with extensive vascular occlusion at the femoropopliteal level. The implantation is carried out via a single inguinal approach in combination with conventional vascular surgical techniques like orthograde ringstripping of the superficial femoral artery, local TEA and patch-grafting of the femoral bifurcation, angioplasty, local lysis, etc. The self-expanding thin-walled PTFE stent prothesis appears to be a viable alternative to the femoropopliteal P1 bypass and can be performed under local anesthesia, percutaneously or through a small inguinal incision. The implantation can be carried out with minimal tissue trauma. PMID- 10460290 TI - [A new treatment method in severe fecal incontinence in children]. AB - We report a case of successful operative therapy of a 12-year-old child with fecal incontinence (Kelly-Holschneider score: 2 points). In the german-speaking area it was the first implantation of an artificial bowel sphincter in a child. The operative procedure, clinical results and anorectal measurements are described. PMID- 10460291 TI - [Benign ulcer in the gastric tube 9 years after esophagectomy for adenocarcinoma]. AB - Esophagectomy for esophageal carcinoma is associated with significant morbidity. Long-term survival rates are usually poor. Common problems in the early postoperative period are pulmonary complications and leakage at the collar anastomosis causing local infection, sepsis and subsequent anastomotic strictures. There are only a few reports in the literature about benign ulcerations in the gastric tube after resection of the esophagus for carcinoma. We report a patient with a huge benign ulcer in the gastric tube penetrating into the right lung 9 years after esophagectomy for carcinoma. Several aspects of possible etiologic factors and different prophylactic approaches as well as therapeutic aspects are discussed. PMID- 10460292 TI - [Hepatic ischaemia during Whipple resection: case report of a rarely required revascularisation]. AB - With the performance of a partial duodenopancreatectomy, the collateral vascular bed between the coeliac trunk and the superior mesenteric artery is reduced. Normally, this does not lead to ischaemia of the upper abdominal organs. However, we present the rare case of a patient in whom a Whipple resection in combination with a hyploplastic coeliac trunk led to hepatic and splenal ischaemia and aorto hepatic bypass grafting. The indication for preoperative arteriography is in discussion, since variations or alteration of the upper abdominal vessels are known to be common but manifest organ ischaemia during resection is a rare complication. Patients with signs of general arteriosclerosis or those in whom upper abdominal resection has been performed previously may benefit from angiography. In other cases a test occlusion before vessel ligation is considered to be suitable in assessing the need for vascular surgical intervention. PMID- 10460293 TI - [Traumatic pulmonary hernia]. AB - The traumatic pulmonary hernia is a rare and uncommon observation. We report a case of a 52-year-old patient who had a car accident. Beside multiple polytraumatic injuries we were able to diagnose an incarcerated pulmonary hernia. Hernia size, incarceration and the respiratory insufficiency of the patient necessitating immediate surgical intervention. We relocated the hernia into the chest and stabilized the thoracic wall. The postoperative course was uneventful. PMID- 10460294 TI - [Solitary fibrous tumors of the pleura]. AB - Solitary fibrous tumors of the pleura (SFTP) are very rare neoplasms. The majority of these tumors are benign, but about 10-20 % fulfill the criteria of malignancy. The clinical presentation varies according to the size and intrathoracic localization. In early stages, often asymptomatic, the tumors may grow to an enormous size and then cause symptoms such as cough, chest pain and dyspnea, but also paraneoplastic syndromes such as hypoglycemia or digital clubbing. Between 1981 and June 1998 we treated in our institution 16 SFTP in 14 patients (4 M, 10 F, average age at first operation 58 years). Eight patients showed symptoms, whereas in the other cases the tumors were found on routine chest X-rays. The usually pedunculated SFTP were completely resected without complications. Two patients developed malignant recurrences, which infiltrated the right upper lung lobe and the diaphragm respectively. In these cases the tumor was resected together with the adjacent structures. Since late recurrences are more often malignant than primary SFTPs long-term follow-up is mandatory even in benign lesions. PMID- 10460295 TI - [Littre's hernia--clinical aspects and review of the history]. AB - In 1700, the French surgeon Alexandre de Littre described for the first time a new form of inguinal hernia. This hernia varied from the known forms of hernias in its clinical course and in the postmortem examination results performed by Littre himself. The characteristic feature of this hernia was the fact that the entire circumference of the bowel wall was not part of the hernial sac, but only the antimesenteric part of the intestinal wall. The underlying pathomechanism was explained 100 years later by Meckel. In a scientific paper about hernias some years earlier, Richter described the intestinal wall hernia, and this initiated the confusing use of the clinical entity known as the Richter-Littre hernia in Germany. In this case report we describe the historic development of this entity. PMID- 10460296 TI - [Surgery of nervous system injuries. I]. PMID- 10460297 TI - [Nail changes in rheumatic disease]. AB - Clinically the nail organ ist easy to investigate. When estimating dermato rheumatological diseases, knowledge of important associated nail symptoms is extremely helpful. In some diseases like osteoarthropathia psoriatica and special variants, Reiter's syndrome and connective tissue disease, typical or even characteristic symptoms can be found. In other disorders like rheumatoid arthritis, goal and systemic vasculitis, associated nail disorders are not uncommon, but of less diagnostic specificity. We review data from the literature and from our dermato-rheumatological outpatient clinic. PMID- 10460298 TI - [Tryptase, a marker for the activation and localization of mast cells]. AB - The serine protease tryptase (ECNr. 3.4. 21.59), which is almost exclusively expressed in mast cells, is released by mast cell degranulation in an enzymatically active form together with other mediators, e.g. histamine, into the extracellular space and the circulation. The capability of the enzyme to directly stimulate several cell types as well as to cleave polypeptide hormones and to activate pro-enzymes suggests a role for tryptase in inflammatory and tissue remodeling processes. Therefore, in the skin, a role of tryptase is suggested not only in mastocytosis and immediate type hypersensitivity reactions, but also in other inflammatory diseases, degenerative or neoplastic conditions as well as in wound healing, where an accumulation and/or activation of mast cells is found. Extracellular tryptase may be superior to histamine as a parameter for the onset and course of immediate type reactions and as an indicator for the activation of mast cells in other conditions. Its absence during histamine-liberating reactions may suggest basophil activation. In addition, tryptase has been shown to be a sensitive and specific marker for the localization of mast cells in tissues. PMID- 10460299 TI - [CO(2) laser vaporisation of actinic cheilitis]. AB - As a premalignant condition, actinic cheilitis requires therapy, and multiple treatment modalities have been described. Although frequently reported in Angloamerican literature, CO(2) laser vaporization for the treatment of actinic cheilitis is scarcely mentioned in German journals. We report 19 cases of CO(2) laser treatment of actinic cheilitis. In only one case, the ablation proved to be too superficial after the first treatment. Except in this case we did not see any recurrences and only one case with minor scarring within a median follow up period of 16 months. In comparison with other treatment modalities, we consider the CO(2) laser ablation to be a safe and cost-saving method with excellent cosmetic results. PMID- 10460300 TI - [Endoscopic fasciotomy and subfascial perforator division for chronic stasis ulcers]. AB - Chronic venous ulcer disease is often refractory to conservative treatment modalities. After surgery of the superficial vein system, endoscopic methods can be used for division of incompetent perforators or to perform paratibial fasciotomy in cases of chronic functional compartment syndromes. We report on 13 endoscopically performed paratibial fasciotomies with or without concomitant endoscopic subfascial division of perforators (ESDP) in patients with stasis ulcers present for a median duration of 15 years. In all patients we observed immediate reduction of pain and edema. In 8 of 13 cases the ulcers healed within 3 months, another ulcer healed within 6 months and the remaining 4 ulcers showed a reduction in size of more than 75%. We conclude that endoscopically performed fasciotomy with or without ESDP is highly effective and has its place in the treatment of chronic venous ulcer disease. PMID- 10460301 TI - [Long term results following radiation therapy of locally recurrent and metastatic malignant melanoma]. AB - The 20-year radiotherapy (RT) experience in patients with locally advanced, recurrent or metastatic malignant melanoma (MM) is analyzed with respect to different endpoints and prognostic factors. From 1977 to 1995, 2917 consecutive patients were entered in our MM registry. RT was indicated in 121 patients (56 females, 65 males) for palliation in advanced MM stages. The histology of the primary lesion was nodular in 51, superficial spreading in 35, acral-lentiginous in 8 and lentigo maligna in 4 patients); 22 were missing or could not be reclassified. Eleven patients had primary or recurrent lesions which were ineligible for surgery or had residual disease (R2) after resection of a primary or recurrent lesion (UICC IIB); 57 patients had lymph node (33) or in-transit metastases (24) (UICC III), 53 had distant organ metastases (7 M1a; 46 M1b) (UICC IV). Time from first diagnosis to on-study RT averaged 19 (median: 18; range: 3 186) months. In most cases conventional RT was applied (2-6 Gy single fractions) up to a mean total RT dose of 45 (median: 48; range: 20-66) Gy. At 3 months follow-up (FU), complete response (CR) was achieved in 7 (64%) and overall response (CR+PR) in all (100%) UICC IIB patients, in 25 (44%)/44 (77%) of 57 UICC III patients, and in 9 (17%)/26 (49%) of 53 UICC IV patients. Progression during RT occurred in 25 (21%) patients. Patients with CR survived longer (median: 40 months) than those without CR (median: 10 months) (p<0. 01). At last FU, 26 patients were alive: 6 (55%) UICC IIB, 17 (30%) UICC III, and 3 (6%) UICC IV patients (p<0.01). In univariate analysis following favorable prognostic factors for CR and long-term survival were identified: low UICC stage (p<0.001), primary site head and neck and total dose >40 Gy (all p<0). PMID- 10460302 TI - [Localized reticulate hyperpigmentation]. AB - 34 year-old pregnant woman presented with reticulate pigmentation of the flexures, the dorsum of the hands and the genitoperianal region. She was in good health and her family history was unremarkable. Histologic examination of the hyperpigmented patches revealed pigmented filiform downgrowths of the interfollicular epidermis and follicular infundibula, as well as small epithelial cysts. Upon immunohistochemical and ultrastructural studies, the number of melanocytes appeared normal. The elongated dendritic processes of the melanocytes contained many mature melanosomes. In the adjacent keratinocytes large melanosomes did not aggregate into complexes. The diagnosis of localized reticulate pigmentary disorder was established. The knowledge of the broad clinical spectrum of localized reticulate hyperpigmentations with its favorable prognosis is of practical importance. Genital or flexural pigmented lesions have to be differentiated from melanosis of the vulva or acanthosis nigricans. The presented case gives further evidence that many of the proposed entities characterized clinically by reticulate pigmented macules and hyperkeratotic follicular lesions are different phenotypic expressions of the same autosomal dominant genodermatosis. PMID- 10460303 TI - [Focal acral hyperkeratosis]. AB - Focal acral hyperkeratosis is a disorder characterized by hyperkeratotic papules along the border of the hands and feet. Focal acral hyperkeratosis and several marginal punctate keratodermas closely resemble acrokeratoelastoidosis of Costa. The question if there is a difference between acrokeratoelastoidosis of Costa and focal acral hyperkeratosis or if they are variants of the same entity are discussed. We report a case showing focal acral hyperkeratosis, review the clinical and histological features of related conditions and present a modified concept of marginal papular acrokeratodermas. PMID- 10460304 TI - [Acatalasemia--Takahara's disease]. AB - The case of a 30 year old man with acatalasemia is presented. The congenital disorder is charaterized by a lack or major reduction of catalase, an enzyme that catalyzes the decomposition of hydrogen peroxide to oxygen and water. The defect is inherited in an autosomal recessive fashion. Occasionally the defect manifests as progressive oral gangrene or Takahara's, disease. PMID- 10460305 TI - [Acanthosis nigricans associated with transitional cell carcinoma of the bladder- symptomatic treatment with calcipotriol]. AB - Acanthosis nigricans is a hyperkeratotic mucocutaneous eruption of heterogenous etiology which is characterized by hyperpigmentation, velvety cutaneous thickening, intensified skin markings and development of verrucous excrescences typically involving the intertriginous areas. Malignant acanthosis nigricans is most often associated with an abdominal adenocarcinoma frequently unresectable at the time of diagnosis. We report on the rare association of acanthosis nigricans with a transitional cell carcinoma of the urinary bladder. Symptomatic treatment with topical calcipotriol significantly reduced the hyperkeratotic and papillomatous skin changes. PMID- 10460306 TI - [Sensitization to gold salts and circumscribed scleroderma]. PMID- 10460307 TI - [Letter from Kismierz-Dolney]. PMID- 10460308 TI - [Tadeusz Chorzelski--(1928-1999)]. PMID- 10460309 TI - [Exanthema in childhood. 2: Bacterial and drug-induced exanthema, exanthema after bone marrow transplantation, exanthema of uncertain etiopathogenesis]. PMID- 10460310 TI - Neue erkenntnisse zum wirkmechanismus von topisch appliziertem vitamin A (all trans-Retinsaure) AB - LANGUAGE="DE">Eine Vielzahl von Untersuchungen hat in den letzten Jahren gezeigt, dass die topische Anwendung von all-trans-Retinsaure, der biologisch aktiven Form des Vitamins A, die Haut gegenuber zwei der wichtigsten durch Sonnenlicht hervorgerufenen Schaden, namlich der Lichtalterung und der Photokarzinogenese, zu schutzen vermag [1].http://link.springer-ny. com/link/service/journals/00105/bibs/50n8p618.html PMID- 10460311 TI - A comparison of the effects of amphetamine and low doses of apomorphine on operant force production, inter-response times and response duration in rat. AB - RATIONALE: Low doses of apomorphine (APO), a non-selective dopamine (DA) agonist, are thought to suppress motor activity via the preferential activation of DA autoreceptors, which effectively reduces DA tone. OBJECTIVES: The suppressant effects on operant responding of low doses of apomorphine were explored and compared with the effects of amphetamine (AMP), an indirect DA agonist. METHODS: In an operant task, rats were trained to press sequentially three separate beams under the following different behavioral requirements: low-force beam (1 g50 g), and a long-duration beam (response duration>2 s). Inter-response times and kinetic measures, such as peak force, the rate of rise of force and response duration, were recorded. Following training, performance was assessed after systemic injection of low doses of APO (0.01, 0.03 and 0.1 mg/kg, s.c.) and AMP (0.1, 0.3 and 1.0 mg/kg, i.p.). RESULTS: APO decreased peak force for the high-force and the long-duration beams by decreasing the rate of rise of force, but did not affect performance success on the low force beam or response duration on the long-duration beam. This indicates that APO impaired the ability to generate high forces but did not interfere with the memory or execution of an overall motor plan. Low doses of APO also increased the times taken to switch from one response to the next and to visit the tray when food was present. In contrast, AMP at 1.0 mg/kg shortened both the time taken to switch between responses and the time spent visiting the food tray. CONCLUSIONS: Low doses of APO interfered with response initiation and execution, suggesting that dopamine acts as a "gating" system, enabling certain processes to be carried out in an efficient and automated manner. PMID- 10460312 TI - Effects of opiate antagonist treatment on the alcohol deprivation effect in long term ethanol-experienced rats. AB - RATIONALE: Opiate antagonists are promising pharmacotherapeutic agents for the treatment of alcohol dependence, reducing craving and relapse rates in weaned alcoholics. However, preclinical findings indicate that they can also increase ethanol consumption and preference in animals with a strong liking for ethanol, depending on the dose and treatment regimen. OBJECTIVE: The present study examined the effects of chronic, intermittent and acute opiate antagonist treatment on the alcohol deprivation effect (ADE) in long-term ethanol- experienced rats, which is an animal model of craving and relapse. METHODS: Long term ethanol-experienced rats were either implanted with mini-osmotic pumps delivering 0, 0.5 or 1 mg/kg per hour naloxone (chronic treatment) or received intermittent naltrexone injections (2x5 mg/kg per day SC). Effects of chronic and intermittent treatment on the ADE were studied in a four-bottle home cage drinking paradigm. In a second experiment, long-term ethanol-experienced rats trained in an operant ethanol self-administration paradigm received acute naltrexone treatment (0, 0.1, 1 or 10 mg/kg SC) before a 23-h session either during basal drinking or during the ADE. RESULTS: Chronic naloxone treatment increased ethanol preference during the ADE. Intermittent naltrexone treatment at a dose comparable to the lower dose of chronic treatment moderately attenuated the ADE. Acute naltrexone treatment selectively reduced lever pressing for ethanol both during the ADE and during basal drinking only at the lowest dose, whereas higher doses also suppressed water intake. The ethanol-specific suppressant effect on the ADE was long lasting. Concerning basal drinking, however, naltrexone had a long lasting reductive effect only on lever pressing for water. CONCLUSIONS: A low dose of naltrexone and an intermittent treatment regimen seem to be necessary to maintain a specific reduction in ethanol intake in individuals with a high motivation to consume ethanol. These findings are consistent with the notion that, at low doses, opiate antagonists reduce the reward value of reinforcers. PMID- 10460313 TI - Pharmacological characterization of the discriminative stimulus properties of the phencyclidine analog, N-[1-(2-benzo(b)thiophenyl)-cyclohexyl]piperidine. AB - RATIONALE: Although both cocaine and the phencyclidine analog, BTCP, have dopamine (DA) re-uptake blocking properties, under some conditions their behavioral effects can be differentiated. Therefore, we examined whether the discriminative stimulus (DS) effects of BTCP are different from those of cocaine. OBJECTIVES: To compare the effects of monoamine re-uptake blockers, varying in their in vitro potencies as inhibitors of DA, norepinephrine (NE), or serotonin re-uptake, in different groups of rats trained to discriminate either BTCP or cocaine from saline. Additionally, drugs from other pharmacological classes were tested in both groups. METHODS: Rats were trained to discriminate either BTCP (5 mg/kg, i.p.) or cocaine (10 mg/kg, i.p.) from saline under a two-lever FR10 drug discrimination procedure. RESULTS: BTCP and cocaine cross-substituted in BTCP- and cocaine-trained rats. The DA re-uptake blockers, mazindol, indatraline, methylphenidate, GBR12909, and GBR12935, occasioned dose-related drug-lever (DL) selection both in cocaine- and in BTCP-trained rats, with potencies that were significantly correlated. In contrast, the NE re-uptake blockers, nisoxetine, desipramine, and nortriptyline, produced higher levels of DL selection in BTCP trained rats than in cocaine-trained rats, a profile like that reported in low dose cocaine-trained rats. Drugs from other classes acted similarly in both discriminations. Further, the alpha1-adrenergic antagonist prazosin dose dependently blocked the DS effects of the training dose of BTCP, but not of cocaine. CONCLUSIONS: The results suggest that the DS effects of BTCP are similar to cocaine, and resemble those of a low training dose of cocaine. PMID- 10460314 TI - Cognitive functioning is susceptible to the level of blood glucose. AB - RATIONALE: It is traditionally assumed that under normal conditions the brain is well supplied with glucose, its basic fuel. However, given the limited stores of glucose in the brain and its dependence on a continual peripheral supply of glucose, it was considered whether the availability of glucose, and the ability to efficiently utilise glucose, affects cognitive functioning. OBJECTIVE: There is increasing evidence that the provision of blood glucose influences memory. To date, the impact of blood glucose on non-memory task performance has received little attention. The present study investigated whether the performance of non memory tasks was susceptible to the level of blood glucose. Two studies are reported in which the influence of a glucose containing drink on six cognitive tests was considered. RESULTS: The consumption of a glucose containing drink resulted in faster performance on the Porteus Maze and greater Verbal Fluency. Higher levels of blood glucose on arrival at the laboratory were associated with better performance on the Water Jars test. With both the Porteus Maze and Block Design tests, after taking a glucose drink, poor performance was associated with blood glucose that remained at higher levels. CONCLUSION: It was suggested that we should consider two physiological mechanisms, firstly, that an equilibrium develops between plasma and brain glucose, such that those with higher levels of blood glucose could be expected to have higher levels of brain glucose and secondly, whether there are individual differences in the efficiency with which glucose is taken from the blood; those with poor glucose control perform some cognitive tasks more poorly. PMID- 10460315 TI - Moderate cortical EEG changes in apolipoprotein E-deficient mice during ageing and scopolamine treatment but not after nucleus basalis lesion. AB - RATIONALE: Recent studies suggest that apoE-deficient mice may have impaired central cholinergic function and neuronal recovery capacity. OBJECTIVE: We investigated whether apoE-deficient mice are more susceptible to the biochemical and EEG defects induced by ageing or nucleus basalis (NB) lesion. METHODS: ApoE deficient and control mice were used. The baseline EEG activity and EEG response to a muscarinic acetylcholine receptor antagonist, scopolamine (0.05 and 0.2 mg/kg) and a benzodiazepine receptor agonist, diazepam (0.5 and 2.0 mg/kg), were studied during ageing. In addition, the cortical and hippocampal ChAT activities were measured in aged mice. The baseline EEG activity and EEG response to scopolamine (0.05 and 0.2 mg/kg), and cortical ChAT activity, were studied after quisqualic acid-induced unilateral NB lesion. RESULTS: The baseline EEG fast wave activity (relative alpha and beta) was higher in apoE-deficient mice. Ageing decreased relative alpha activity similarly in both strains. The scopolamine induced EEG slowing was less prominent in apoE-deficient than in control mice, and the difference between the strains became slightly clearer during ageing. The NB lesion failed to produce more severe changes in cortical EEG and ChAT activity in apoE-deficient mice. Cortical and hippocampal ChAT activity was equal in young and aged apoE-deficient and control mice. The EEG response to diazepam in young and aged mice was similar in both strains. CONCLUSIONS: The regulation of cortical EEG activity of apoE-deficient mice was somewhat altered during ageing and the response to scopolamine treatment was blunted. However, the cholinergic cells of the NB of apoE-deficient mice were not more sensitive to lesion or to ageing, suggesting that apoE does not have to be present to preserve the viability of cholinergic neurons. PMID- 10460316 TI - Effect of novel environmental stimuli on rat behaviour and central noradrenaline function measured by in vivo microdialysis. AB - RATIONALE: Although physically aversive stimuli induce functional changes in central noradrenergic neurones, little is known about the noradrenergic response to environmentally aversive stimuli. OBJECTIVES: The first aim was to characterise environmental features that are perceived as stressful by rats. The second was to investigate whether changes in the concentration of extracellular noradrenaline are induced by these environmental features. METHODS: A light/dark shuttle-box was used to test rats' behavioural response to a range of stimuli (novelty, bright light, and the presence of an unfamiliar rat), either before or after microdialysis probe implantation. Changes in the concentration of extracellular noradrenaline in the frontal cortex and hypothalamus in vivo were then evaluated on exposure to these same test conditions. RESULTS: Naive rats spent less time in a brightly-lit test arena than a dark one. However, the behavioural response to the light arena was attenuated by the presence of an unfamiliar rat. Probe implantation intensified the response to the light arena but did not affect behaviour in the dark arena. In the microdialysis studies, there was no change in the concentration of extracellular noradrenaline on transfer of rats to the dark arena but there was an increase in both the frontal cortex (+45%) and hypothalamus (+75%) on exposure to the light arena. A similar increase was induced in both brain regions when the light arena contained an unfamiliar rat. CONCLUSIONS: Implantation of a microdialysis probe modifies the behavioural responses to certain environmental stimuli. Regardless of this, the extent to which rats perceive a novel environment as aversive is not the only determinant of the noradrenergic response to such stimuli. However, differences in stimulus controllability in the microdialysis and the behavioural experiments could influence the apparent intensity of the stress. PMID- 10460317 TI - Idazoxan potentiates rather than antagonizes some of the cognitive effects of clonidine. AB - Several investigations have revealed substantial influences of pharmacological manipulation of central noradrenergic activity upon performance in cognitive tests sensitive to frontal lobe dysfunction. They suggest a significant role for the noradrenergic coeruleo-cortical projection in cognitive function but conflicting findings and the complex pharmacology of adrenoceptor agents make it difficult to be precise about underlying mechanisms. In order to clarify these we have compared the effects of an alpha1/alpha2-adrenoceptor agonist, clonidine, an alpha2-adrenoceptor antagonist, idazoxan, and these agents in combination. Three groups of healthy volunteers were used to investigate the effects of these noradrenergic manipulations upon performance of tasks from the CANTAB test battery known to be sensitive to frontal lobe dysfunction. Previously reported effects of clonidine upon sustained visual attention and upon session-to-session improvement were replicated. Furthermore, idazoxan inhibited the hypotensive effect of clonidine. Idazoxan had no overall effect on performance of any of the tests but did inhibit session-to-session improvement in performance of a planning task, attentional set shifting and sustained visual attention. Rather than leading to the anticipated mutual antagonism of effects, combining clonidine and idazoxan led to a wider and more striking range of cognitive impairments. These results are discussed alongside findings which support a role for imidazoline (I1) receptors in blood pressure control, where clonidine and idazoxan are antagonistic, and evidence of less potent antagonism at somato-dendritic alpha2 adrenoceptors in the locus coeruleus. PMID- 10460318 TI - Manipulations of mu-opioid and nicotinic cholinergic receptors in the pontine tegmental region alter cocaine self-administration in rats. AB - RATIONALE: The pedunculopontine tegmental nucleus (PPTg) has been implicated in drug reward, particularly in the development of dependence. However, little is known of the receptor systems within this nucleus which might be involved. Furthermore, some research suggests that the PPTg may also be part of the neuronal circuitry involved in established drug-taking behavior. OBJECTIVE: The objective of these experiments was to examine the role of mu-opioid and nicotinic cholinergic mechanisms in the PPTg in cocaine self-administration. METHODS: Microinfusions of mu-opioid and nicotinic receptor selective compounds were made into the PPTg of rats trained to self-administer cocaine intravenously, in the vicinity of cholinergic cells which are known to project to the midbrain dopamine neurons of the ventral tegmental area (VTA). RESULTS: The mu-opioid selective agonist DAMGO, tested at doses of 0, 0.05 and 0.5 microg, produced a dose-related reduction in the number of cocaine infusions obtained during the 1-h self administration sessions. The mu-selective antagonist CTOP (0-2 microg) and nicotine (0-10 microg) did not produce significant changes in cocaine self administration. Microinfusions of the nicotinic antagonist dihydro-beta erythroidine (0-30 microg) produced a small but significant increase in cocaine maintained responding. CONCLUSIONS: These data show that mu-opioid mechanisms in the PPTg can influence cocaine self-administration markedly. Moreover, the data demonstrate that PPTg circuitry can influence drug reward in already-established drug-reinforced behavior, as well as during the development of dependence (as shown by previous research). PMID- 10460319 TI - Effects of training dose and two- versus three-choice testing procedure on nicotine discrimination responding in humans. AB - RATIONALE: Discrimination of a drug's interoceptive stimulus effects often depends substantially on training and testing conditions. OBJECTIVES: We examined changes in nicotine discrimination behavior in humans as a function of lowering the training dose and of varying the discrimination testing procedure. METHODS: Smokers and never-smokers (n=10 each) were initially trained to discriminate 20 microg/kg nicotine by nasal spray from placebo (0) and tested on generalization of discrimination responding across a range of doses from 0 to 20 microg/kg. Each subsequently learned to reliably discriminate progressively smaller doses of nicotine from placebo until his or her threshold dose for discrimination was identified (mean=2.7 microg/kg). A repeat testing of generalization responding across 0-20 microg/kg was then conducted, using placebo and the subject's threshold dose as training doses. Generalization testing involved both two-choice and three-choice (novel response option) quantitative procedures. RESULTS: A significant shift to the left was seen in nicotine-appropriate responding in the two-choice procedure when the nicotine training dose was lowered (i.e. from the first to the second test of generalization). In the three-choice procedure, however, there was no such leftward shift. Instead, in never-smokers, a flattening of nicotine-appropriate responding occurred with a lowering of the training dose, while novel-appropriate responding significantly increased. The subjective effects of "head rush" and, in never-smokers only, "jittery" also showed a shift to the left in their relationship with nicotine generalization dose when the training dose was lowered. CONCLUSIONS: These results confirm the importance of training and testing conditions on discrimination behavior and subjective drug responses within subjects and demonstrate the utility of the novel-response, three-choice procedure for assessing qualitatively different stimulus effects of novel drug doses. PMID- 10460320 TI - Context-specific morphine tolerance on the paw-pressure and tail-shock vocalization tests: evidence of associative tolerance without conditioned compensatory responding. AB - RATIONALE: Demonstrations of associative tolerance to the analgesic effects of morphine, not confounded by practice or novelty effects, have been restricted to the tail-flick and flinch-jump tests. OBJECTIVES: Experiment 1 investigated whether associative tolerance would be found on two other nociceptive assessment methods: the paw-pressure withdrawal and tail-shock vocalization thresholds. Experiment 2 tested the hypothesis that conditioned compensatory behavioral responses are the substrate of associative morphine tolerance in the paw pressure, tail-shock, and tail-flick tests. METHODS: Rats were given eight morphine injections (20 mg/kg, i.p.) explicitly paired or unpaired with a distinctive context. Control animals were given saline injections over the course of conditioning. Animals were then tested after morphine (experiment 1) or placebo injections (experiment 2) in the context. RESULTS: There was evidence of context-specific tolerance across both testing methods, with a rightward shift of dose-response curves of paired relative to unpaired animals. No evidence of conditioned compensatory responding was found on any of the three testing methods. CONCLUSIONS: The data indicated that, although Pavlovian processes can play a major role in tolerance acquisition, there was little support for the thesis that the conditioned tolerance response is a behavioral effect that is opposite in direction to the direct effects of the drug. PMID- 10460321 TI - Nicotine blocks quinpirole-induced behavior in rats: psychiatric implications. AB - RATIONALE AND OBJECTIVES: Because of known and imputed roles of dopaminergic and nicotinic cholinergic systems in a variety of neurological and neuropsychiatric disorders, combined neurochemical and behavioral methods assessments were made to study the intermodulatory roles of these neurochemical systems. METHODS: Rats were treated daily during postnatal ontogeny with the dopamine D2/D) agonist, quinpirole (QNP) HCl (1.0 mg/kg/day), for the first 3 weeks from birth. This priming process replicated previous findings of behavioral sensitization, manifested as hyperlocomotion, increased paw treading with jumping, and increased yawning. RESULTS: All effects were partially or totally blocked by acute treatment with nicotine (0.3 mg/kg, i.p.). The effects of nicotine, in turn, were partially or totally blocked by the nicotinic antagonist, mecamylamine (1.0 mg/kg, i.p.). In concert with these behavioral actions, QNP-primed rats displayed greater binding of [3H]cytisine in midbrain and cerebellum and greater [125I]alpha-bungarotoxin binding in hippocampus and striatum. CONCLUSIONS: Accordingly, these selective ligands for alpha4beta2 and alpha7 nicotinic receptors, respectively, demonstrate that nicotinic receptors are altered by dopamine D2/D3 agonist treatment of rats with primed dopamine receptors. We propose that nicotinic agonists may have a therapeutic benefit in behavioral disorders brought about by central dopaminergic imbalance. PMID- 10460323 TI - The effects of alcohol cues and an alcohol priming dose on a multi-factorial measure of subjective cue reactivity in social drinkers. AB - RATIONALE: Exploring subjective alcohol cue reactivity in non-clinical samples should assist understanding in clinical samples where additional problems muddy the water. However, exploration is stalled through using insensitive, single-item representations. OBJECTIVE: The effect of alcohol cues and a priming dose of alcohol on a new multi-factorial representation of cue reactivity is sought (DAQ, Desire for Alcohol Questionnaire). METHODS: Prime and Cue exposure are variables in a standard 2x2 between subjects design set within a stooge taste-evaluation experiment. The DAQ was administered after a Prime and Cue exposure phase. RESULTS: Main effects for Cue exposure but not Prime were found for the DAQ total and the subscales Mild desires (positively reinforcing items) and Strong desires/intentions but not Negative reinforcement (negatively reinforcing items) and Controllability; however, there was no interaction. CONCLUSION: The DAQ is a sensitive measure of subjective cue reactivity in social drinkers and its potential in the evaluation of pharmacological interventions is proposed. PMID- 10460322 TI - Acute and chronic effects of nornicotine on locomotor activity in rats: altered response to nicotine. AB - RATIONALE: Nicotine, a tobacco alkaloid, is known to be important in the acquisition and maintenance of tobacco smoking. Nornicotine, an active nicotine metabolite, stimulates nicotinic receptors and may produce psychomotor effects similar to nicotine. OBJECTIVE: The present study determined the effects of acute and repeated administration of nornicotine on locomotor activity and compared its effects with those of nicotine. METHODS: R(+)-Nornicotine (0.3-10 mg/kg), S(-) nornicotine (0.3-10 mg/kg), S(-)-nicotine (0.1-1 mg/kg) or saline was administered s.c. to rats acutely or repeatedly (eight injections at 48-h intervals). Activity was recorded for 50 min immediately after each injection. RESULTS: S(-)-Nicotine produced transient hypoactivity, followed by dose-related hyperactivity. Repeated S(-)-nicotine administration resulted in tolerance to the hypoactivity and sensitization to the hyperactivity. Subsequent testing following a saline injection revealed evidence of conditioned hyperactivity. Acute administration of 0.3 mg/kg or 1 mg/kg R(+)- or S(-)-nornicotine produced no effect. Transient hypoactivity was observed at 3 mg/kg and 10 mg/kg R(+) nornicotine and at 10 mg/kg S(-)-nornicotine. However, rebound hyperactivity was not observed following acute administration of either nornicotine enantiomer, suggesting that nornicotine-induced psychomotor effects differ qualitatively from those of S(-)-nicotine. Repeated R(+)-nornicotine resulted in tolerance to the transient hypoactivity, however hyperactivity was not observed. Repeated S(-) nornicotine resulted in tolerance to the hypoactivity and the appearance of hyperactivity. Repeated administration of either nornicotine enantiomer resulted in a dose-dependent alteration in response to a 1 mg/kg S(-)-nicotine challenge, suggesting some commonalities in the mechanism of action. CONCLUSION: Nornicotine likely contributes to the neuropharmacological effects of nicotine and tobacco use. PMID- 10460324 TI - Letter from the editor PMID- 10460325 TI - Imaging of sickle cell disease. AB - Sickle cell disease is an important health care issue in the United States and in certain areas in Africa, the Middle East and India. Although a great deal of progress has been made in understanding the disease at the molecular and pathophysiologic level, specific treatment which is safe and accessible for most patients is still elusive. Going into the next millennium, the management of this disease is still largely dependent on early diagnosis and the treatment of complications with supportive care. Thus, diagnosis and evaluation of the complications of the disease are crucial in directing clinical care at the bedside. Modern imaging modalities have greatly improved, and their application in the patient with the sickling disorders has enhanced the decision - making process. The purpose of this article is to review the clinical aspects of common complications of the disease and to discuss imaging approaches which are useful in their evaluation. PMID- 10460326 TI - Cervicomedullary astrocytomas of childhood: clinical and imaging follow-up. AB - BACKGROUND. Cervicomedullary astrocytomas are a unique subset of brainstem tumors in children because they have a good prognosis when compared to the pontine subset of brainstem gliomas. Objective. To review the clinical and imaging findings in a series of children with cervicomedullary astrocytomas as to diagnosis and management. MATERIALS AND METHODS: A retrospective review of eleven children (six females, five males, age range: 10 days-18 years; mean = 7 years) with cervicomedullary tumors was done including the clinical presentation, imaging studies (MR: eleven, CT and MR: four), surgical findings, pathological results, and follow-up clinical and imaging findings (range: 0.2-11 years; mean = 5.2 years). RESULTS: Symptoms and signs were delayed and protracted, often occurring over months to years (mean = 2.3 years, range 0.5-7 years). The tumors expanded the dorsal medulla and involved the upper cervical spinal cord (mean maximum tumor diameter = 4.4 cm). Only three patients had hydrocephalus. In three of four cases the tumor was not seen on CT. On MR, the majority of the tumors were T1 hypointense and T2 hyperintense. Treatment consisted of surgery only in six patients, surgery and radiation therapy in four, and surgery, chemotherapy, and radiation in one. There was recurrent local disease in four patients and on follow-up metastatic disease in the brain in one. On follow-up the majority of the patients are alive and stable (mean = 5.2 years, range 0.2-11 years). There has been one death. The majority of tumors were pilocytic astrocytomas. CONCLUSION: Cervicomedullary tumors are a unique subset of brainstem gliomas in childhood that present with a long duration of symptoms and a greater long-term survival than pontine gliomas. PMID- 10460327 TI - MRI appearances of metachromatic leukodystrophy. AB - BACKGROUND: The leukodystrophies constitute a wide spectrum of cerebral disorders of varying etiology. The imaging appearances on CT and MRI are recognizable as abnormalities of white matter; however, it may be impossible to arrive at the correct diagnosis based on imaging studies alone. PATIENTS AND METHODS: Three patients of varying age and clinical symptomatology diagnosed with metachromatic leukodystrophy (MLD) had remarkably similar MRI appearances. A "tigroid" or "leopard-skin" appearance was demonstrated within deep white matter in each case. RESULTS: All of the patients had biochemical confirmation of MLD. CONCLUSION: Although the "tigroid" pattern previously was considered to be pathognomonic of Pelizaeus-Merzbacher disease, the diagnosis of MLD must now be considered when these MRI appearances are encountered. PMID- 10460328 TI - Rickets on MR images. AB - BACKGROUND: The pathologic changes at the physis in patients with rickets have been well demonstrated histologically. Radiographs can depict only the associated osseous abnormalities. PATIENTS AND METHODS: We report two children in whom MR imaging demonstrated rachitic changes in the physeal cartilage beyond the well recognized bony features. RESULTS: The striking appearance of the physes and the physes of the secondary ossification centers confirm that MR imaging can successfully evaluate the cartilaginous structures of the developing skeleton. CONCLUSION: Though MR imaging is clearly unnecessary for the diagnosis of rickets, it is important that the typical features are not misinterpreted as other pathology. PMID- 10460329 TI - The association between Turner's syndrome and juvenile rheumatoid arthritis. AB - BACKGROUND: Turner's syndrome (TS) is a chromosomal disease frequently associated with autoimmune conditions including thyroid disease, inflammatory bowel disease, and diabetes. Recent reports have described an association with juvenile rheumatoid arthritis (JRA) and psoriatic arthritis. We describe three additional cases of TS associated with JRA. OBJECTIVE: The objective of this report is to describe the radiographic and clinical features of TS associated with JRA to heighten awareness of this association and alert radiologists to recognize the superimposition of radiographic bony changes of chronic joint disease and the bone changes of TS. Patients and methods. Clinical history and radiographic images of three girls with TS and arthritis were reviewed. The radiographic findings typical of TS and juvenile arthritis are described. RESULTS: Of about 65 patients at our center with Turner's syndrome 3 had JRA (as described in this report), supporting the association between TS and JRA. All our patients who met American College of Rheumatology Classification criteria for JRA had radiographic and clinical findings consistent with both their JRA and Turner's syndrome. CONCLUSION: We believe that it is important to consider the diagnosis of Turner's syndrome in girls with JRA, recognizing that characteristic radiographic findings such as metacarpal shortening are usually present. Conversely, suspicion of an underlying inflammatory arthritis is warranted in search for radiological findings consistent with JRA in girls with TS and joint symptoms. PMID- 10460330 TI - Skeletal abnormalities in fetuses with Down's syndrome: a radiographic post mortem study. AB - OBJECTIVE: To evaluate skeletal abnormalities on post-mortem radiographs of fetuses with Down's syndrome. MATERIALS AND METHODS: Biometrical and morphological criteria, which are used for US prenatal detection of trisomy 21, were assessed. Limb long bones, biparietal diameter (BPD)/occipito-frontal diameter (OFD) ratio, ossification of nasal bones and appearance of the middle phalanx of the fifth digit (P2) in 60 fetuses with Down's syndrome were analysed and compared with 82 normal fetuses matched for gestational age (GA) from 15 to 40 weeks' gestation (WG). RESULTS: We observed reduced growth velocity of limb long bones during the third trimester in both groups, but the reduction was more pronounced in the trisomic group. Brachycephaly was found as early as 15 WG in Down's syndrome and continued throughout gestation (sensitivity 0.28, specificity 1). Ossification of the nasal bones, which can be detected in normal fetuses from 14 WG, was absent in one quarter of trisomic fetuses, regardless of GA. The middle phalanx of the fifth digit was evaluated by comparison with the distal phalanx (P3) of the same digit. We found that P2 was not ossified in 11/31 trisomic fetuses before 23 WG, and was either not ossified or hypoplastic in 17/29 cases after 24 WG (sensitivity 0.56, specificity 1). CONCLUSIONS: Three key skeletal signs were present in trisomic fetuses: brachycephaly, absence of nasal bone ossification, and hypoplasia of the middle phalanx of the fifth digit. All these signs are appropriate to prenatal US screening. When present, they fully justify determination of the fetal karyotype by amniocentesis. PMID- 10460331 TI - Hepatic enhancement analysis in children using smart prep monitoring for 2 : 1 pitch helical scanning. AB - PURPOSE: To analyze hepatic enhancement by using Smart Prep protocols appropriate for children of different weight groups and 2:1 pitch helical CT imaging as the investigative tools. PATIENTS AND METHODS: A group of 55 children ranging in weight between 20 and 180 lbs underwent 67 contrast-enhanced abdominal helical CT examinations using Smart Prep (GE Medical Systems, Milwaukee, Wisc.). Of these studies, 21 (31 %) were excluded because of failure to follow the prescribed Smart Prep protocols. Smart Prep protocols were established for nine different weight groups. Scan delay, aorta and liver time to peak, and liver enhancement over baseline were recorded. RESULTS: Optimal abdominal CT studies with adequate contrast enhancement of hepatic and portal veins were obtained in 46 patients. There was no significant difference in the time between peak aortic and the liver enhancement among different weight groups (mean time 12.0 +/- 7.1 s for all children). However, the mean hepatic enhancement over baseline in children weighing < 30 lbs was below 50 Hounsfield units (HU) compared to the rest of the children who had mean hepatic enhancement of > 50 HU. CONCLUSION: Two-thirds of the Smart Prep protocols were successfully implemented, and all of these resulted in good contrast enhancement of hepatic and portal veins. Optimal mean liver enhancement (> 50 HU) was seen in children >/= 30 lbs. Children < 30 lbs had mean liver enhancement of 33 HU +/- 7.2 above the baseline likely caused by contrast dose. PMID- 10460332 TI - MR urography: the future gold standard in paediatric urogenital imaging? AB - BACKGROUND: Examination of the paediatric urogenital tract is traditionally performed using methods that utilise ionising radiation, such as intravenous urography (IVU), computerised tomography (CT), voiding cystourethrography (VCU), and scintigraphy, in addition to ultrasound (US). OBJECTIVE: To determine the potential and effectiveness of MR urography (MRU) in infants and children. Materials and methods. 44 MRU examinations were prospectively performed in 39 patients (21 infants, mean age 3.5 months, and 18 children, mean age 6 years 2 months) with known or suspected pathology of the urinary tract. Non-enhanced, fast spin-echo sequences (TSE) were performed in all patients. In 70 % of the patients a contrast-enhanced, fast gradient-echo sequence (TFE) was included. The dynamic sequence was prolonged and supplemented with furosemide provocation in some patients with suspected urinary-tract obstruction. RESULTS: Nine percent of examinations were non-diagnostic or interrupted due to movement. MRU contributed additional information in 66 %. Nine patients with suspected urinary-tract obstruction were examined with both contrast-enhanced MRU and scintigraphy. Three MRU examinations were less informative and one equal to scintigraphy when obstruction was the diagnosis. When using a technique with a prolonged dynamic sequence, including frusemide provocation, four MRU examinations were equal and one was superior to scintigraphy. CONCLUSIONS: MRU has the potential to replace traditional diagnostic methods which use ionising radiation in paediatric patients. Further studies are needed before definite conclusions can be drawn. PMID- 10460333 TI - Nonradiopaque penetrating foreign body: "a sticky situation". AB - Foreign bodies within soft tissues are common in children. They may cause a chronic inflammatory reaction that can result in abnormal findings on radiographs, including lytic or blastic osseous changes. These radiographic findings can mimic both benign and malignant processes. In cases where the history is uncertain and the foreign body is not recognized, magnetic resonance (MR) imaging can make a specific diagnosis and direct appropriate therapy. PMID- 10460334 TI - Ultrasound features of intussusception predicting outcome of air enema. AB - OBJECTIVE: To examine features identified on US which predict success or failure of air-enema reduction of intussusception. MATERIALS AND METHODS: A retrospective study of 117 consecutive episodes of intussusception, presenting for US over a 6 year period. The specific features examined were: free fluid within the peritoneum, small-bowel obstruction, colonic wall thickness, and fluid trapped between the colon and the intussusceptum. RESULTS: The overall reduction rate, irrespective of US features, over the 6-year period was 72 %. Reduction rates were significantly higher with the absence of free fluid, trapped fluid, or small bowel obstruction (93 %). The presence of trapped fluid predicted an unfavourable outcome, with a significantly lower success rate (25 %). Colonic wall thickness did not predict outcome; in successful reductions, mean wall thickness was 7.2 mm and in failed reductions 7.6 mm. CONCLUSIONS: Where free fluid, small-bowel obstruction, and trapped fluid are absent, almost 100 % success with air-enema reduction should be achievable. Where trapped fluid is present, air enema should be performed cautiously to avoid perforation caused by overvigorous attempts at pneumatic reduction of an incarcerated intussusception. PMID- 10460335 TI - Imaging of pediatric mesenteric abnormalities. AB - The relative paucity of mesenteric fat seen in the pediatric population can make detection and localization of processes in the mesentery difficult. This pictorial essay reviews pediatric mesenteric disorders and presents criteria that help localize processes to the mesentery. Disorders are categorized by specific patterns of involvement, which can readily be identified by imaging: developmental abnormalities of mesenteric rotation, diffuse mesenteric processes, focal mesenteric masses, and multifocal mesenteric masses. PMID- 10460336 TI - Abnormal clavicles in a neonate with partial monosomy 21 by Wang and Aftimos, New Zealand. PMID- 10460337 TI - Editorial commentary on Roebuck DJ:"Risk and benefit in paediatric radiology". PMID- 10460338 TI - Incoherent neutron scattering of copper azurin: a comparison with molecular dynamics simulation results. AB - The low-frequency dynamics of copper azurin has been studied at different temperatures for a dry and deuterium hydrated sample by incoherent neutron scattering and the experimental results have been compared with molecular dynamics (MD) simulations carried out in the same temperature range. Experimental Debye-Waller factors are consistent with a dynamical transition at approximately 200 K which appears partially suppressed in the dry sample. Inelastic and quasielastic scattering indicate that hydration water modulates both vibrational and diffusive motions. The low-temperature experimental dynamical structure factor of the hydrated protein shows an excess of inelastic scattering peaking at about 3 meV and whose position is slightly shifted downwards in the dry sample. Such an excess is reminiscent of the "boson peak" observed in glass-like materials. This vibrational peak is quite well reproduced by MD simulations, although at a lower energy. The experimental quasielastic scattering of the two samples at 300 K shows a two-step relaxation behaviour with similar characteristic times, while the corresponding intensities differ only by a scale factor. Also, MD simulations confirm the two-step diffusive trend, but the slow process seems to be characterized by a decay faster than the experimental one. Comparison with incoherent neutron scattering studies carried out on proteins having different structure indicates that globular proteins display common elastic, quasielastic and inelastic features, with an almost similar hydration dependence, irrespective of their secondary and tertiary structure. PMID- 10460339 TI - Poisson-Boltzmann model studies of molecular electrostatic properties of the cAMP dependent protein kinase. AB - Protonation equilibria of residues important in the catalytic mechanism of a protein kinase were analyzed on the basis of the Poisson-Boltzmann electrostatic model along with a cluster-based treatment of the multiple titration state problem. Calculations were based upon crystallographic structures of the mammalian cAMP-dependent protein kinase, one representing the so called closed form of the enzyme and the other representing an open conformation. It was predicted that at pH 7 the preferred form of the phosphate group at the catalytically essential threonine 197 (P-Thr197) in the closed form is dianionic, whereas in the open form a monoanionic ionization state is preferred. This dianionic state of P-Thr197, in the closed form, is stabilized by interactions with ionizable residues His87, Arg165, and Lys189. Our calculations predict that the hydroxyl of the Ser residue in the peptide substrate is very difficult to ionize, both in the closed and open structures of the complex. Also, the supposed catalytic base, Asp166, does not seem to have a pK(a) appropriate to remove the hydroxyl group proton of the peptide substrate. However, when Ser of the peptide substrate is forced to remain ionized, the predicted pK(a) of Asp166 increases strongly, which suggests that the Asp residue is a likely candidate to attract the proton if the Ser residue becomes deprotonated, possibly during some structural change preceding formation of the transition state. Finally, in accord with suggestions made on the basis of the pH-dependence of kinase kinetics, our calculations predict that Glu230 and His87 are the residues responsible for the molecular pK(a) values of 6.2 and 8.5, observed in the experiment. PMID- 10460340 TI - Mathematical simulation of chlorophyll a fluorescence rise measured with 3-(3',4' dichlorophenyl)-1,1-dimethylurea-treated barley leaves at room and high temperatures. AB - Chlorophyll a fluorescence induction (FI) measured by Plant Efficiency Analyser fluorometer at room temperature shows a typical O-J-I-P pattern which is at high temperature changed to an O-K-P pattern with a new step K. It has been suggested that the appearance of the K step reflects inhibition of an oxygen evolving complex (OEC). When FI is measured at room temperature with the photosystem II (PSII) herbicide 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU), which blocks electron transport from Q(A) to Q(B) (the first and the second quinone electron acceptors in PSII, respectively), the time course of the FI shows a sigmoidal increase to the maximal fluorescence which is reached at a little longer time than that of the J step. Similarly, the FI measured at high temperature with DCMU reaches the maximal value of fluorescence at the time which is a little longer than that of the K step. On the other hand, the reversible radical pair model (RRP) describes energy utilization and electron transport up to Q(A). In this work we present the first, to our knowledge, RRP model extended by a description of the function of the donor side of PSII. Assuming the inhibition of the OEC or its full function, the extended RRP model successfully simulates the fluorescence rise measured with DCMU at high and room temperatures, respectively. The roles of the initial state of the OEC and the values of the rate constants in the extended RRP on the simulations of the fluorescence rise at room and high temperatures are also discussed. PMID- 10460341 TI - Constructing the suitable initial configuration of the membrane-protein system in molecular dynamics simulations. AB - A method for constructing the suitable initial configuration of the membrane protein system for molecular dynamics (MD) simulations is presented. This method could provide some hydrated initial configurations and help us to determine the best surface area of the system by contracting the surface area and comparing the optimized lowest energy of the system by energy minimization. The gramicidin A (GA) channel in;the fully hydrated dimyristoylphosphatidylcholine (DMPC) bilayer was used as our model. Three configurations with different surface areas were selected and applied for one 400 ps and two 300 ps MD simulations at constant pressure and temperature. All simulations were fairly stable without any constraints. Through analysis of the MD trajectories we found that the system with the best surface area was more stable than the other two systems, whose sizes were changed in the simulations. Further analysis of the bilayer normal length and the order parameters of the lipid alkyl tails indicates that the system with the best surface area shows some characteristics of the L(alpha) phase, while both the smaller and the larger size systems have distinct deviations from the L(alpha) phase that we expect. This illustrates that the correct surface area and the suitable initial configuration have an important influence on the phase of the membrane in the MD simulation. In addition, by comparing the root mean square differences of GA relative to the initial structure and interaction energy between different components of the system for all three systems, we find that the state of the DMPC bilayer has exerted a significant influence on the structure of GA. All these results demonstrate the validity of our method for constructing the initial configuration of the membrane protein system for MD simulations. PMID- 10460342 TI - Simulation of the packing of idealized transmembrane alpha-helix bundles. AB - The aim of this study is to investigate if the packing motifs of native transmembrane helices can be produced by simulations with simple potentials and to develop a method for the rapid generation of initial candidate models for integral membrane proteins composed of bundles of transmembrane helices. Constituent residues are mapped along the helix axis in order to maintain the amino acid sequence-dependent properties of the helix. Helix packing is optimized according to a semi-empirical potential mainly composed of four components: a bilayer potential, a crossing angle potential, a helix dipole potential and a helix-helix distance potential. A Monte Carlo simulated annealing protocol is employed to optimize the helix bundle system. Necessary parameters are derived from theoretical studies and statistical analysis of experimentally determined protein structures. Preliminary testing of the method has been conducted with idealized seven Ala(20) helix bundles. The structures generated show a high degree of compactness. It was observed that both bacteriorhodopsin-like and delta endotoxin-like structures are generated in seven-helix bundle simulations, within which the composition varies dependent upon the cooling rate. The simulation method has also been employed to explore the packing of N = 4 and N = 12 transmembrane helix bundles. The results suggest that seven and 12 transmembrane helix bundles resembling those observed experimentally (e.g., bacteriorhodopsin, rhodopsin and cytochrome c oxidase subunit I) may be generated by simulations using simple potentials. PMID- 10460343 TI - Molecular dynamics simulation of alpha-melanocyte stimulating hormone in a water membrane model interface. AB - The conformation of the tridecapeptide alpha-melanocyte stimulating hormone in the presence of a double water-membrane interface was studied by molecular dynamics simulation, using the computational package THOR. In this program the solvent is represented by a continuous medium with dielectric constant epsilon, and the interface between different media is simulated by a surface of discontinuity of the dielectric constant. The electrostatic image method was used to write down the terms, added to the force field, that describe the polarisation effects induced in the interface by the atomic charges. The program was further improved by the introduction of a second surface, parallel to the first one, to mimic the membrane. A conformational search using the software Prelude was employed to find an initial geometry for the peptide in water. The molecular dynamics simulation performed during 10 ns showed that the peptide structure is flexible in water, without stabilisation of any preferential conformation. In the presence of the model membrane, the peptide moved to the medium representing the interior of the membrane. Inside the low dielectric constant medium, the structure of the peptide showed a turn in the central sequence of amino acids and a packed conformation remained stabilised during more than 7.0 ns of simulation. PMID- 10460344 TI - Calculation of hydrodynamic properties of macromolecular bead models with overlapping spheres. AB - For the calculation of hydrodynamic properties of rigid macromolecules using bead modelling, models with overlapping beads of different sizes are used in some applications. The hydrodynamic interaction tensor between unequal overlapping beads is unknown, and an oversimplified treatment with the Oseen tensor may introduce important errors. Here we discuss some aspects of the overlapping problem, and explore an ad hoc form of the interaction tensor, proposed by Zipper and Durchschlag. We carry out a systematic numerical study of the hydrodynamic properties of a two-spheres model, showing how the Zipper-Durchschlag correction removes efficiently the numerical instabilities, and predicts the correct limits. PMID- 10460345 TI - Topography of cell traces studied by atomic force microscopy. AB - Migrating adherent cells release material onto artificial substrates like glass and silicon while moving. Traces of mouse fibroblasts (L929) have been visualised by atomic force microscopy (AFM). "Non-contact" mode AFM in a liquid environment can extract topographic information from these traces. This dynamic mode allows the study of these soft structures without damage or compression. The AFM images show crossing and branching networks (with specific angles of branching), structured patches, nodular elements, linear elements with irregular height and other features. Fourier analysis of segment spacing in the strands is presented. These spatial features of fibroblast traces are strong indications that actin linked to structural proteins is involved in the formation of cell traces. We also give methods for trace preparation and undistorted imaging and discuss further perspectives. PMID- 10460346 TI - Thermal stability of a flavoprotein assessed from associative analysis of polarized time-resolved fluorescence spectroscopy. AB - Upon gradually heating a particular mutant of the flavoprotein NADH peroxidase, it was found from the peculiar time-resolved fluorescence anisotropy pattern of the flavin prosthetic group (FAD) that, at elevated temperature, FAD is released from the tetrameric enzyme. Since in this case a mixture of free and enzyme-bound FAD contributes to the time-dependent fluorescence anisotropy, its analysis can only be accomplished by an associative fitting model, in which specific fluorescence lifetimes of both species are linked to specific correlation times. In this letter the general approach to the associative polarized fluorescence decay analysis is described. The procedure can be used for other flavoproteins to determine the temperature at which the onset of thermal denaturation will start, leading to release of the flavin prosthetic group. PMID- 10460347 TI - Treatment-related chronic myelogenous leukemia. AB - Treatment-related (Tr) AML and MDS after chemotherapy, radiotherapy, or the combination of both have been well characterized. However, tr-CML seems to differ from these better-known entities in frequency, clinical course, and prognosis. Tr CML cannot be distinguished from de novo CML cytogenetically, and, in contrast to tr-AML and tr-MDS, typical chromosomal aberrations related to tr-CML have not been described. Treatment-related CML is a late effect of cytotoxic or immunosuppressive therapy which might be increasingly recognized due to a higher number of patients treated with intensive therapy regimens. We review here the available data on incidence of tr-CML as well as the affected individual's characteristics with regard to different treatment options in malignant and nonmalignant diseases. PMID- 10460348 TI - Prevalence study and molecular characterization of alpha-thalassemia in Filipinos. AB - In order to determine the prevalence and molecular basis of alpha-thalassemia (thal) among Filipinos, a total of 2954 Filipinos in Taiwan were enrolled in this study. A complete blood count was done for every subject. Those with microcytosis (MCV less than 82.5 fl) were studied with hemoglobin (Hb) high-performance liquid chromatography to determine the levels of Hb A2 and Hb F, and with an enzyme immunoassay to determine plasma ferritin levels. Those who had microcytosis and normal or low levels of Hb A2 and Hb F were further studied with molecular methods for alpha-globin gene mutations. We used Southern blot hybridization and/or the polymerase chain reaction to detect Southeast Asian deletion, Filipino deletion, rightward and leftward single alpha-globin gene deletions, and Hb Constant Spring and Hb Quong Sze. Specific amplification and direct DNA sequencing of the alpha2- and alpha1-globin genes were carried out in apparent alpha-thal carriers without any of the above-mentioned mutations. Our results showed that in Filipinos the prevalence of alpha-thal 1 was 5% (147 carriers) and that of alpha-thal 2 was 1.7% (49 carriers); two had Hb H disease. Among the alpha-thal 1 carriers, 89 had the Southeast Asian deletion and 58 had the Filipino deletion. Among the alpha-thal 2 carriers, 48 had a rightward deletion and one had a leftward deletion. None had Hb Constant Spring or Hb Quong Sze. Specific amplification and DNA sequencing in five apparent alpha-thal carriers did not reveal mutations in the 2-kb region spanning the alpha2- and alpha1 globin genes. The molecular defects of alpha-thal in Filipinos were different from those in the neighboring ethnic groups. Elucidation of the alpha-thal mutations in Filipinos is useful in the genetic counseling and prenatal diagnosis of this common disease. PMID- 10460349 TI - Serum transferrin receptors in detection of iron deficiency in pregnancy. AB - A prospective hospital-based study was conducted to evaluate the efficacy of serum transferrin receptors in the detection of iron deficiency in pregnant women. The iron status of 100 pregnant women with single uncomplicated term pregnancies in the first stage of labor was established using standard laboratory measures. These included complete hemogram, red cell indices, serum iron, percent transferrin saturation, and serum ferritin. In addition, serum transferrin receptor (STFR) was estimated. The results of 81 women with complete laboratory profiles were analyzed. Thirty-five (43.2%) women were anemic (hemoglobin <11 g/dl). Hemoglobin (Hb) showed a significant correlation with MCH, MCHC, serum iron, and percent transferrin saturation, suggesting that the anemia was likely to be due to iron deficiency. The mean STFR level was 18.05+/-9.9 mg/l in the anemic women and was significantly raised (p<0.001) compared with that of the nonanemic women. STFR correlated significantly with Hb (p<0.001), MCH (p<0.05), MCHC (p<0.01), serum iron (p<0.01), and percent transferrin saturation (p<0.01) and also showed a highly significant correlation with the degree of anemia. Serum ferritin in these women did not correlate with Hb, and only 54.4% of the women had levels <12 ng/ml, which does not reflect the true prevalence of iron deficiency. Serum transferrin receptor estimation is thus a useful measure for detecting iron deficiency in pregnancy. PMID- 10460350 TI - Comparison of different methods for separation and ex vivo expansion of cord blood progenitor cells. AB - Umbilical cord blood is capable of hematopoietic stem cell reconstitution in children. However, the major limitation of cord blood is a relatively low content of pluripotent progenitor cells. Thus, safe engraftment for adolescents and for adults is still not predictable and a technology for ex vivo expansion of umbilical cord blood cells is desirable. In a first step, four different methods of red cell depletion followed by magnetic cell sorting of CD34+ cells were evaluated in this study in order to assess the efficacy and safety of optimal stem cell recovery. A modified two-step Ficoll gradient separation and a hydroxyethyl starch separation tended to produce a better WBC/MNC recovery (median 94.2+/-2.44% vs. 90.2+/-5. 8%) as compared with standard Ficoll gradient separation and a gelatin-based procedure (median 78.35+/-7.1% vs. 67.2+/-5.5%). However, the recovery of CD34+ cells after magnetic cell sorting did not reach a statistically significant difference after the four different methods of red cell depletion, indicating that the recovery of WBC/MNC is not predictably correlated with the recovery of stem cells within these fractions. In a second step, we established three different cytokine combinations by adding the megakaryocyte growth and development factor +/- erythropoietin and granulocyte colony stimulating factor to a fetal calf serum containing medium with Flt 3, stem cell factor, and interleukin-3. Net expansion of total colony-forming cells 20- to 50 fold and expansion of colony-forming cells after 5 weeks of culture 1.5- to 3 fold were obtained over a period of 7-14 days. These results demonstrate that cord blood stem cells can be expanded substantially in this short-term culture system. PMID- 10460351 TI - Assessment of renal function in patients with multiple myeloma: the role of urinary proteins. AB - Renal failure (RF) in multiple myeloma (MM) is considered an ominous complication even though, when timely therapy is started in patients with minimal damage, a high percentage of cases can achieve a regression. The evaluation of renal involvement usually relies on serum creatinine or its clearance, but these parameters have proved to be inadequate to identify initial damage. The aim of this study was to assess the role of the following urinary proteins in diagnosing renal impairment at an early stage: high-molecular-mass proteins (transferrin, IgG, albumin) as markers of glomerular damage, and low-molecular-weight proteins and parenchymal enzymes [alpha(1)-acid glycoprotein (AGP), alpha(1)-microglobulin (alpha(1)M), retinol-binding protein (RBP), beta(2)-microglobulin (beta(2)M), lysozyme (LZ), and N-acetyl-beta-d-glucosaminidase (NAG)] as indicators of tubular disorder. Thirty MM patients (nine at disease onset and 21 previously treated) were included in the study. No correlation was found between the urinary proteins and the phase or the stage of the disease. By the Spearman test, Bence Jones proteinuria correlated significantly with the 24 h proteinuria (p=0. 01) and beta(2)M (p=0.02), and weakly with the alpha(1)M. Serum creatinine concentrations and urea correlated with most of the analytes evaluated: RBP correlated well with urea (p=0.004) and creatinine (p=0.004); IgG (p=0.006) albumin (p=0.009), AGP (p=0.04), and NAG (p=0.02) correlated with serum creatinine. Significant statistical correlation was found between all the analytes except LZ and the creatinine clearance. Twelve of the 30 MM patients (40%) showed abnormal values of urinary proteins. Four of these patients showed overt renal failure with significant modification of the serum parameters and of creatinine clearance, three showed an isolated decrease of creatinine clearance, and five did not present any alteration of serum or urinary parameters. This testifies to the utility of urinary proteins in highlighting renal damage even in cases where the customary serum indicators of renal disorder are normal. In conclusion, our results demonstrate that AGP, RBP, NAG, transferrin, and IgG are good indicators of renal damage. They do not correlate with the severity of the disease, but they seem to be helpful in identifying a subset of patients with initial renal dysfunction. PMID- 10460352 TI - Hematopoietic donor chimerism and graft-versus-myeloma effect in relapse of multiple myeloma after allogeneic bone marrow transplantation. AB - A large group of patients relapsing after allogeneic bone marrow transplantation (BMT) have obtained remission after infusion of leukocytes from their original donor, suggesting a graft-versus-myeloma effect. However, side effects such as graft-versus-host disease and myelosuppression are severe, and sometimes fatal, complications of this therapeutic approach. Previously we demonstrated that patients with leukemia who lack donor hematopoiesis in relapse after BMT experience severe and lasting aplasia after infusion of donor leukocytes. In two patients - one with extramedullary and one with marrow relapse after a sex mismatched transplantation - we analyzed hematopoietic chimerism by cell sorting and bone marrow cultures. CD34-positive cells, CD4-CD8-positive cells, committed progenitors, and LTC-IC were of donor origin, as demonstrated by two-color fluorescence in situ hybridization (FISH). Additionally, in relapse complete donor T-cell chimerism was seen. In contrast, plasma cells were of recipient origin in the patient who had a relapse in the bone marrow. Both patients were treated with infusions of donor leukocytes from their original donor. Neither patient suffered myelosuppression, and one achieved a stable complete remission. PMID- 10460353 TI - Fludarabine, cytarabine, and G-CSF (FLAG) for the treatment of acute myeloid leukemia relapsing after autologous stem cell transplantation. AB - Twenty-six patients affected by acute myeloid leukemia (AML) who relapsed after autologous stem cell transplantation (ASCT) were treated with the FLAG regimen (fludarabine, cytarabine, and G-CSF). Their median age was 39 years (range 14 59). The median interval from achievement of CR to ASCT was 4 months (2-8). The conditioning regimen was BAVC (BCNU, amsacrine, VP-16, cytarabine) in eight patients, BuCy (busulfan, cyclophosphamide) in 13, and TBI-Cy (total body irradiation, cyclophosphamide) in five. Relapse occurred after a median of 7 months (2-18). ASCT had been performed in CR1 for 23 patients and in CR2 for three. Nineteen patients had been given bone marrow, seven peripheral blood stem cells collected following consolidation plus G-CSF. Overall, CR was obtained by 13 patients (50%), all remitters requiring a single course. The median time for hematological recovery of neutrophils >500/microl and platelets >20,000/microl was 24 and 30 days, respectively. The median duration of G-CSF administration was 25 days, while the median hospitalization was 31 days. There were four deaths in induction (15%), while nine patients (35%) were resistant. After achieving CR, two patients received allogeneic BMT, five a second ASCT, and four were consolidated with HD-ARA-C. Only two patients were judged unable to receive any further therapy. There were 14 documented infections, while nine patients experienced fever of unknown origin. WHO >2 nonhematological toxicity consisted of stomatitis (50%), hepatic dysfunction (11%), diarrhea (11%), and lethargy (4%). Median overall survival and disease-free survival were 6 and 13 months, respectively. Six patients are in CCR at present. We conclude that FLAG is effective in patients with AML who are relapsing after ASCT. The toxicity is acceptable, enabling most patients to receive further treatment, including second transplantation procedures. PMID- 10460354 TI - Disseminated nocardiosis as a complication of Evans' syndrome. AB - Nocardiosis is an opportunistic infection caused by gram-positive, weakly acid fast filamentous aerobic organisms. Three species cause infection in man: N. asteroides, N. brasiliensis, and N. caviae, the first one being the most common. With increased use of immunosuppressive therapy for various autoimmune diseases, opportunistic infection by Nocardia has increasingly been reported. N. asteroides infections manifest in various ways; the lungs, skin, and brain are the organs most frequently involved. We describe a patient with Evans' syndrome, a disease requiring long-term immunosuppression, who acquired systemic nocardiosis. The infection was primarily pulmonary, misdiagnosed as tuberculosis, with subsequent hematogenous dissemination to the skin and central nervous system. The diagnosis of cerebral involvement was difficult to prove, as the patient presented with stroke-like episodes. After a positive blood culture was obtained, antibiotic therapy was introduced. The patient's condition deteriorated and the brain with infiltration of the meninges, lungs, skin, and kidneys. Nocardia is an important but often overlooked opportunistic infectious agent in immunocompromised hosts, causing diagnostic and therapeutic problems. As the mortality of cerebral nocardiosis is greater than 80%, early diagnosis and appropriate therapy are crucial. PMID- 10460356 TI - MR imaging of the head and neck. AB - The purpose of this paper is to describe current imaging protocols for MR imaging of the head and neck region and to define results and clinical impact. Depending on the clinical question, different MRI protocols are presented for imaging of the head and neck. The appearance of different pathologic findings on imaging studies and how adapted imaging protocols help to improve differential diagnosis is discussed. In summary, MRI is the method of choice for imaging of the head and neck. PMID- 10460357 TI - Spinal cord tumors. AB - Spinal cord tumors are rare; however, every radiologist should be able to recognize and readily identify those lesions often found in younger patients or children [1, 2, 3, 4, 5, 6, 7, 8,9]. Early diagnosis plays an important role in the management of the lesions and interferes with the prognosis and final outcome of the patient [10]. Plain X-ray of the spine and CT are of limited diagnostic value in cases of intramedullary pathology. Magnetic resonance imaging should be performed as soon as possible and as the first technique whenever an intrinsic spinal cord lesion is clinically suspected. Systematic analysis of the MR images together with a basic knowledge of the most common spinal cord tumors encountered, including astrocytomas, ependymomas, and hemangioblastomas, should provide a correct diagnosis and lead to appropriate treatment subsequently. PMID- 10460358 TI - MR imaging of the spine: trauma and degenerative disease. AB - The purpose of this paper is to discuss the capabilities and drawbacks of MR imaging in patients with trauma to the spine and degenerative spinal conditions. In spinal trauma MR imaging is secondary to plain X-ray films and CT because of the greater availability and ease of performance of these techniques and their superior capability for detecting vertebral fractures. Magnetic resonance imaging is useful for detecting ligamentous ruptures and intraspinal mass lesions such as hematoma, and for assessing the state of the spinal cord and prognosis of a cord injury. In degenerative spinal disease the necessity is emphasized of critically evaluating the clinical relevance of any abnormal feature detected, as findings of degenerative pathology are common in individuals without symptoms. Magnetic resonance myelography permits rapid and accurate assessment of the state of the lumbar nerve roots (compressed or not). In the cervical region the quality of the myelographic picture is often degraded in patients with a narrow spinal canal. PMID- 10460359 TI - Non-invasive evaluation of the visceral arteries with magnetic resonance angiography. AB - Traditionally, there have been only a few indications for imaging of the visceral arteries. With improvements in treatment of many mesenteric vascular disorders, it has become more important to establish a diagnosis early in the course of the disease. With the advent of ultrasound, computed tomography and magnetic resonance imaging non-invasive imaging of the visceral arteries became possible, although all these modalities were limited in scope for demonstration of mesenteric pathology. The advent of high-quality mesenteric MR arteriography and venography, which allows comprehensive evaluation of both visceral artery anatomy and function, has led to a huge increase in the number of studies directed at the mesenteric arteries in many departments. PMID- 10460360 TI - MR angiography: supra-aortic vessels. AB - Magnetic resonance angiography (MRA) has become an imaging modality which comprises various techniques based on two concepts: methods relying on the natural flow effects, the time-of-flight and phase-contrast technique, either in two- or three-dimensional acquisition mode, and the more recently developed contrast-enhanced (CE) MRA methods. The main indications for evaluation of the supra-aortic vessels are, firstly, the grading of carotid artery stenoses caused by an atherosclerotic process, and secondly, the evaluation of dissections of the cervical arteries because this disease plays an important role as a cause for stroke especially in younger patients. The various MRA techniques in their application to the main pathologies encountered at the supra-aortic vessels are presented, and recent developments in the promising field of CE MRA are discussed. PMID- 10460361 TI - MR angiography of run-off vessels. AB - Magnetic resonance angiography has taken a huge step forward since the introduction of contrast-enhanced MR angiography using gadolinium chelates. The more conventional MR angiographic techniques, such as time-of-flight and phase contrast MR angiography, have been ousted by contrast-enhanced MR angiography in most vascular areas. However, in imaging the lower extremities, the major obstacle is the length of the vascular tree. In order to cover the entire peripheral vasculature, at least two to three fields of view are required. Using contrast-enhanced MR angiography, the best results are obtained if the vessels of interest are imaged during passage of a bolus of contrast material. Vessel-to background contrast in subsequent acquisitions using subsequent injections of contrast material is hampered by recirculation and leakage of previously injected gadolinium, enhancing both the venous system and surrounding tissue. To overcome this problem several research groups have come up with various solutions. The three main strategies employed can be classified as either bolus catch, bolus chase, or bolus track techniques. The purpose of this article is to explain working mechanisms of the three bolus imaging strategies for imaging both inflow and outflow vessels of the lower extremities, to show their advantages and disadvantages, and to review results described in the literature in imaging patients using these techniques. PMID- 10460362 TI - Bronchiolar inflammatory diseases: high-resolution CT findings with histologic correlation. AB - Bronchiolar diseases pose a significant challenge to the clinician confronted with the evaluation and management of the affected patient. A variety of infectious and non-infectious diseases may affect the bronchioles causing either reversible or fixed bronchiolar obstruction. High-resolution CT (HRCT) is currently the best imaging modality for evaluation of small-airway disease. In fact, a wide spectrum of abnormalities are identified at HRCT in patients with bronchiolar diseases. These abnormalities are shown on HRCT in the presence of a normal or unclear chest radiograph. Additionally, HRCT performed at suspended full expiration may demonstrate the physiologic consequences of bronchiolar disease, e. g., air trapping. The differential diagnosis of pulmonary manifestations of bronchiolar diseases at HRCT is based on the different patterns of abnormality. Familiarity with the presentation of different bronchiolar inflammatory processes aid the radiologist in narrowing the differential diagnosis or even in suggesting a specific diagnosis. This article reviews the HRCT findings of various bronchiolar inflammatory diseases outlining their pathologic features. Knowledge of the underlying gross and microscopic pathologic features leads to a better understanding of their CT appearances. PMID- 10460363 TI - Chest wall infiltration by lung cancer: value of thin-sectional CT with different reconstruction algorithms. AB - The aim of this investigation was to evaluate whether thin-sectional CT with different reconstruction algorithms can improve the diagnostic accuracy with regard to chest wall invasion in patients with peripheral bronchogenic carcinoma. Forty-one patients with intrapulmonary lesions and tumor contact to the thoracic wall as seen on CT staging underwent additional 1-mm CT slices with reconstruction in a high-resolution (HR) and an edge blurring, soft detail (SD) algorithm. Five criteria were applied and validated by histological findings. Using the criteria of the intact fat layer, HRCT had a sensitivity of 81 % and a specificity of 79 %, SD CT had a sensitivity of 96 % and a specificity of 78 %, and standard CT technique had a sensitivity of 50 % and a specificity of 71 %, respectively. Regarding changes of intercostal soft tissue, HRCT achieved a sensitivity of 71 % and a specificity of 96 %, SD CT had a sensitivity of 94 % and a specificity of 96 % (standard CT technique: sensitivity 50 % and specificity 96 %). For the other criteria, such as pleural contact area, angle, and osseous destruction, no significant differences were found. Diagnostic accuracy of chest wall infiltration can be improved by using thin sectional CT. Especially the application of an edge-blurring (SD) algorithm increases sensitivity and specificity without additional costs. PMID- 10460364 TI - The value of pulmonary angiography for the differential diagnosis of pulmonary embolism. AB - The aim this study was to evaluate potential additional information of pulmonary angiography in patients with suspected pulmonary embolism and non-diagnostic lung scan findings. In a series of 150 patients who underwent pulmonary angiography for suspected venous thromboembolism, the images for potential alternative diagnoses other than pulmonary embolism were evaluated. All patients had non diagnostic lung scan findings. Angiography was performed both by conventional and by digital subtraction angiography techniques. Images were evaluated by at least two experienced readers. Angiograms were scored for both presence or absence of pulmonary embolism, as well as other diagnoses. Pulmonary embolism was proven in 40 patients (27 %) and excluded in 105 patients (70 %), whereas non-interpretable images were obtained in 5 patients (3 %). A range of alternative diagnoses were detected by angiography: atelectasis (n = 24), pleural effusion (n = 15), pneumonia (n = 11), emphysematous bullae (n = 8), neoplasm (n = 3), atrial septum defect (n = 2), chronic thromboembolism (n = 1), and other diagnoses (n = 3). Overall, only 54 patients (36 %) had completely normal angiograms. Although pulmonary angiography remains the reference method for the diagnosis of pulmonary embolism, one has to be aware of other, often unexpected, but equally important findings which could influence the management of the patient. This aspect of pulmonary angiography has been insufficiently emphasized in the literature. PMID- 10460365 TI - Percutaneous CT-guided lung biopsy: sequential versus spiral scanning. A randomized prospective study. AB - The aim of this study was to evaluate in a prospective and randomized study spiral versus sequential scanning in the guidance of percutaneous lung biopsy. Fifty thoracic lesions occurring in 48 patients were biopsied by a senior and a junior operator. Six different time segments of the procedure were measured. Scanning mode versus length of procedure, pathological results, irradiation and complications were evaluated. Total duration of the procedure and of the first sampling was significantly longer with spiral CT for the senior operator (p < 0.004). No significant time difference was observed for the junior operator. Diameter of the lesion, depth of location, position of the patient and needle entry site did not influence the results. The sensitivity was 90.9, specificity 100, positive predictive value 100 and negative predictive value 60 % for spiral CT, and 94.7, 100, 100 and 85.7 % for sequential CT, respectively. Eleven pneumothoraces and ten perinodular hemorrhages were seen with spiral CT and six and ten, respectively, with sequential CT. The mean dose of irradiation was 4027 mAs for spiral CT and 2358 mAs for conventional CT. Spiral CT does neither reduce procedure time nor the rate of complications. Pathological results do not differ compared with sequential CT, and total dose of irradiation is higher with spiral scanning. PMID- 10460366 TI - "Air bubble": a new diagnostic CT sign of perforated pulmonary hydatid cyst. AB - The greatest difficulty in the CT diagnosis of perforated pulmonary hydatid cyst (PPHC) is the increase in the attenuation numbers following infection. Because of the solid density of infected hydatid cysts, the differentiation from an abscess or neoplasm is usually impossible. The aim of this study was to evaluate the value of "air bubble" as a new CT sign in the diagnosis of PPHC. Sixty-five patients (28 men and 37 women) with PPHC were included in the study. As a control group, 55 patients who had malignant (n = 36) or non-malignant (n = 19) pulmonary diseases were also examined. Radiological diagnosis with classical CT findings was made in only 38 of 65 patients (58.5 %) with PPHC. Air bubble sign was positive in 54 of the patients with PPHC (sensitivity 83.1 %) but only 3 of 55 patients in control group (specificity 94.5 %). When we analyzed the CT scans with classical CT findings including air bubble, the diagnosis of PPHC was made in 61 of patients (93.8 %). It is concluded that "air bubble sign" is a valuable CT finding in the diagnosis of PPHC. PMID- 10460367 TI - Intralobar lung sequestration with systemic coronary arterial supply. AB - Pulmonary sequestration is a rare anomaly. An accurate pre-operative evaluation of its vascular supply is essential for the surgeon's operative approach. We describe here an intrapulmonary sequestration with vascular arterial supply via the left circumflex and the right coronary artery. This case demonstrates that if aortography is unrevealing, then a coronary source should be considered in the preoperative search for the arterial supply to a pulmonary sequestration. Moreover, pulmonary sequestration should be listed in the differential diagnosis of aberrant coronary arteries. PMID- 10460368 TI - Dilatation of the rete testis: ultrasound study. AB - The aim of this study was to show the US findings in mode B and color Doppler duplex of the dilatation of the rete testis, in order to analyze its association with other scrotal processes and to confirm their inclusion into the benign testicular lesions. We present seven diagnosed cases (mean age 61 years) of dilatation of the rete testis to which a clinical control and US was accomplished up to 1 year. The scrotal sonography study was carried out with a linear probe of 7.5 Mhz. In the US examination we observed in all cases an intratesticular image located in the mediastinum testis constituted by anechoic and serpiginous tubular structures, which do not show any blood flow with the color Doppler. In one case the mentioned observations were bilateral. Five cases had cysts in epididymis and the last case showed an increase in size in the epididymis head. Color Doppler duplex examination did not detected flow dots in the seven patients. The dilatation of the rete testis is a benign entity frequently associated with pathology in epididymis, with specific US findings which permit avoidance of invasive tests. PMID- 10460369 TI - Anatomic variation in the origin of the main renal arteries: spiral CTA evaluation. AB - The aim of this study was to provide quantitative data on the origin and trajectory of the main renal arteries using spiral CT angiography and arteriography. Normal renal artery anatomy was assessed on spiral CT angiography (axial transverse sections and shaded-surface-display reconstructions) in 100 patients referred for renal arteriography who had no significant renal artery stenosis. Two hundred major renal arteries were studied. The vast majority of right (88 %) and left (87 %) renal arteries originated between the lower third of the first lumbar vertebra and the lower border of the second lumbar vertebra. In 50 patients both ostia were at the same level; in the remaining 50 patients, the right ostium was located above the left in 37 patients. On the right, the angle of origin varied from -10 to + 55 degrees (mean + 24 degrees ). On the left, the angle of origin varied from + 30 to -55 degrees (mean -11 degrees ). Spiral CT angiography provides additional anatomic data, notably regarding the angle of origin of the renal arteries, that is potentially useful for planning interventional procedures. PMID- 10460370 TI - Sclerosing stromal tumor of the ovary: radiologic findings. AB - Sclerosing stromal tumor is a rare ovarian neoplasm. We describe the radiologic findings of sclerosing stromal tumor in two patients. In both patients, MR and CT images showed a large mass in the left adnexal region. On dynamic contrast enhanced images, the tumors showed early peripheral enhancement with centripetal progression. PMID- 10460371 TI - Pediatric liver neoplasms: a radiologic-pathologic correlation. AB - Only 1-2 % of all pediatric tumors occur in the liver. Two thirds of these tumors are malignant and almost all of the tumors cause clinical symptoms due to their mass effects. Besides the poor prognosis in most of the malignant tumors, for further treatment the origin and nature of the neoplasm has to be known. Due to the mostly unimpeded growth into the peritoneal cavity, the origin of the tumors is primarily often unclear and can non-invasively only be determined by advanced imaging techniques. The display of the macro- and microhistological key features of primary pediatric liver neoplasms, including hepatoblastoma (HB), infantile hemangioendothelioma (IHE), mesenchymal hamartoma (MH), undifferentiated (embryonal) sarcoma (UES), and hepatocellular carcinoma (HCC), together with their imaging representation by ultrasound, computed tomography, and magnetic resonance imaging, may deepen the understanding of the underlying pathology and its imaging appearance. Furthermore, in many cases sufficient information may be provided not only to differentiate benign from malignant tumors, but also to guide for adequate treatment. PMID- 10460372 TI - The spectrum of imaging in Currarino triad. AB - The aim of this study was to evaluate the spectrum of findings and the efficacy of different imaging modalities in order to formulate recommendations for diagnostic imaging of Currarino triad (ASP syndrome), including screening of relatives. The imaging films of five female patients (age range 6 weeks to 12 months) were analysed retrospectively. The studied material consisted of US and MRI of the lower spine (5 patients each), lumbosacral plain radiography (4 patients), contrast enema (4 patients), urinary US (2 patients), genitography (1 patient) and myelo-CT (1 patient). Depiction of pathological findings with different imaging modalities was reviewed and validated with special respect to their demonstrability by US. Ultrasonography detected the sacral bony defect as well as the presacral pathology (meningocele and/or tumour) and thereby gave the basic diagnosis in all of the cases. It also depicted tethered cord and urinary tract abnormalities correctly. Magnetic resonance imaging gave a more distinct visualization of pre- and intraspinal pathology with additional demonstration of intraspinal lipoma in two cases. Regarding anorectal and genital malformations, radiographic contrast agent studies had been used in all patients. Two blind ending retrorectal fistulas, depicted by enema, were missed by MRI. Patients with congenital or early infancy obstipation, anorectal malformations and complex urinary tract malformations should have spinal and pelvic sonography first. A plain film of the sacrum is recommended in equivocal cases. The need for MRI and contrast agent studies depends on the individual pathology, whereas presently MRI has made further radiographic imaging increasingly dispensable. A screening program with lumbosacral US or plain radiography for families with Currarino triad should be obligatory. PMID- 10460373 TI - Papillary cystic neoplasm of the pancreas in a teenage boy. AB - We present a case of a 13-year-old boy with a left-sided abdominal mass which proved to be a papillary cystic neoplasm of the pancreas. This low-grade malignant lesion of young patients is very rare, and exceedingly rare in males. The prognosis following resection of this tumor is good. We present the ultrasound and computed tomographic picture of this lesion, as well as the gross and microscopic pathology. PMID- 10460374 TI - Role of hemolysis in potassium release by iodinated contrast medium. AB - It has been demonstrated that an iodinated contrast medium (CM) causes release of potassium into blood vessel lumina, resulting in an increase in serum potassium. The purpose of the present study was to assess whether this potassium release is due to hemolysis. Fresh human blood was mixed in vitro with CM at a ratio of 10:2. Potassium release rates were determined, and serum haptoglobin and free hemoglobin were measured after 30 min of exposure to CM. To compare the potassium release curve between CM exposure and true hemolysis induced by distilled water, fresh human blood was also mixed with distilled water. The level of serum haptoglobin decreased due to hemodilution. Changes in haptoglobin were not correlated with potassium release rates. The serum free hemoglobin level did not increase significantly, and there was no correlation between changes in the free hemoglobin level and the rate of potassium release. Hemolysis caused by water occurred instantaneously, whereas potassium release caused by CM was a slow response, which was linearly correlated with exposure time. Potassium release from blood cannot be explained by hemolysis. PMID- 10460375 TI - Iodixanol in cerebral computed tomography: a randomized, double-blind, phase-III, parallel study with iodixanol and iohexol. AB - Iodixanol is a new nonionic dimer, isotonic with blood at all clinically relevant concentrations. Iodixanol (270 mg I/ml) was compared in a double-blind, randomized, parallel-group, phase-III study to the monomeric nonionic iohexol (300 mg I/ml) for evaluation of safety, tolerability and radiographic efficacy during cerebral CT. One hundred adult patients scheduled to undergo contrast enhanced cerebral CT were randomly allocated to receive either iodixanol or iohexol. All completed the trial. Safety was evaluated by recording discomfort and other adverse events, tolerance by assessing intensity and incidence of discomfort. Radiographic efficacy was assessed from the diagnostic information and the radiographic density. No serious adverse events occurred. One patient (2 %) in the iodixanol group and one patient (2 %) in the iohexol group experienced a transient reddening at the neck and lower neck-line, respectively. Both contrast agents were well tolerated. One patient (2 %) in the iodixanol group and two patients (4 %) in the iohexol group experienced a sensation of warmth (discomfort) in connection with the injection. No difference between the two contrast media were noted radiographically. This comparison between iodixanol and iohexol showed both contrast media to be safe, well-tolerated and efficacious for use in cerebral CT. PMID- 10460376 TI - Imaging findings in patients with myelofibrosis. AB - The purpose of this review is to illustrate the wide range of radiological abnormalities in myelofibrosis. Myelofibrosis, also called myeloid metaplasia, is a myeloproliferative disorder of unknown etiology. The common imaging findings in patients with myelofibrosis are osteosclerosis, hepatosplenomegaly, and lymphadenopathies. In addition, extramedullary hematopoiesis may develop in multiple sites such as chest, abdomen, pelvis, and central nervous system, simulating malignant disease. Selected plain-film, CT, and MR images in patients with myelofibrosis are shown as pictorial essay to allow ready recognition of the most common imaging abnormalities of the disease. PMID- 10460377 TI - Magnetic resonance imaging of lateral epicondylitis of the elbow with a 0.2-T dedicated system. AB - The purpose of this study was to evaluate MR imaging findings of the common extensor tendon in patients with lateral epicondylitis and asymptomatic volunteers studied on a 0.2-T dedicated system. In 23 patients (age range 29-58 years, mean age 47 years) with clinical symptoms of lateral epicondylitis MR imaging was performed using T1-, T2- and contrast-enhanced T1-weighted spin-echo sequences. In addition, the elbows of seven healthy volunteers (age range 22-29 years, mean age 25 years) and the symptom-free contralateral elbow of 11 of the 23 patients (age range 29-58 years, mean age 47 years) were studied as controls. Five patients were surgically treated after the MR examination and the results of histopathology were correlated with MR findings. Of the patients, 95.6 % showed intratendinous signal intensity changes on T1-weighted images on the symptomatic side. In 69.6 % signal alterations were observed on T2-weighted sequences and in 56.5 % an intratendinous contrast enhancement was present. Histopathology showed fibrovascular proliferation and fatty degeneration in patients with distinct signal intensity changes and contrast enhancement. Patients with only minor signal intensity changes on T1- and T2-weighted sequences and no contrast enhancement demonstrated fibrosclerotic degeneration and intratendinous cartilage formation in histopathology. The contralateral elbow showed signal intensity changes in 6 of 11 (54.5 %) cases on T1-weighted images and in 3 of 11 (27.3 %) on T2-weighted images. In the group of healthy volunteers minor signal intensity changes of the common extensor tendon could be seen in only 1 case. In patients with lateral epicondylitis of the elbow the type and extent of pathologic changes within the common extensor tendon can be evaluated using a dedicated low-field MR system. On the basis of MR imaging findings a more specified therapy planning among the variety of treatment modalities can be achieved. PMID- 10460378 TI - Patient throughput times for orthopedic outpatients in a department of radiology: results of an interdisciplinary quality management program. AB - The purpose of this project was to employ quality management methods in order to decrease throughput times for orthopedic outpatients sent to the department of radiology. The following intervals were measured at the onset of the study and after 6 and 12 months: (a) between arrivals at outpatient clinic and radiology counter; (b) between arrival at radiology counter and time of last radiograph; and (c) between time of last radiograph and radiology report printing time. After the initial measurement, numerous changes were initiated both in radiology and in orthopedic surgery. The mean interval between arrival at the outpatient clinic and in radiology decreased by one third from 60 min during the first measurement to 40 (p < 0.001) and 41 min during the second and third measurement. The proportion of patients with total radiology times of more than 30 min decreased from 41 to 29 % between the first and third measurements (p < 0.001). The corresponding results for radiology times of more than 45 min were 17 and 11 % (p = 0.03). A standard type of quality management program can be employed successfully in order to reduce radiology throughput times for orthopedic outpatients. PMID- 10460379 TI - Craniocervical artery dissection: MR imaging and MR angiographic findings. AB - Dissection of the carotid and vertebral arteries is a not so uncommon cause of stroke and has to be considered as a differential diagnosis especially in younger patients. Therapeutic and prognostic implications are different from those in extracranial atherosclerotic disease. Dissection results from hemorrhage into the vessel wall usually between the layers of the media. Digital subtraction angiography (DSA) depicts the resulting luminal compromise that may reveal some typical, but not specific, findings. The same is true for non-invasive angiographic techniques such as time-of-flight magnetic resonance angiography (MRA) and computed tomography angiography (CTA), which have shown accurate results compared with DSA. The main advantage of these techniques is the direct visualization of the vessel wall confirming the intramural hematoma. This is achieved best with MR imaging due to the high signal of blood degradation products on T1- and T2-weighted images. Therefore, MRI in combination with MRA is presently the method of choice for initial diagnosis and follow-up of craniocervical artery dissection (CCAD). In some questionable cases, CTA is a non invasive alternative that is independent of flow phenomena. PMID- 10460380 TI - Echo-planar magnetic resonance imaging (EPI) with high-resolution matrix in intra axial brain tumors. AB - The aim of this study was to assess the potential of high-speed interleaved echo planar imaging (EPI) to achieve diagnostic image quality comparable to T2 weighted imaging in patients with brain tumors. Seventeen patients with intra axial, supratentorial tumors (10 untreated gliomas, 7 radiated gliomas) were investigated on a 1. 5-T scanner. The conventional scan (SE, TR/TE = 2200/80 ms, 18 slices) was acquired in 8 min, 4 s, and EPI (TR/TE = 3000/80 ms, 18 slices) was completed in 25 s. The films were compared in a blinded trail by three radiologists. On the general impression and anatomic display, both sequences were rated to be of similar quality. Artifacts were slightly more pronounced at the skull base and around surgical clips using EPI. Tumor delineation was nearly equivalent using EPI, compared with the T2-weighted sequence. Echo-planar imaging reached diagnostic quality in all patients. Interleaved high-resolution EPI yielded sufficient quality to depict intra-axial, supratentorial brain tumors. Since EPI can be obtained in a small fraction of the time needed for conventional spin echo, in addition to other indications it could be considered to study patients unable to cooperate. PMID- 10460381 TI - Lymphocytic hypophysitis presenting with diabetes insipidus: MR findings. AB - It has been thought that neurohypophysial involvement manifesting as central diabetes insipidus in lymphocytic hypophysitis is rare. The radiological and clinicopathological features of two cases represent a variant of lymphocytic adenohypophysitis and/or lymphocytic infundibulo-neurohypophysitis are discussed. PMID- 10460382 TI - Idiopathic hypertrophic cranial pachymeningitis associated with an orbital pseudotumor. AB - We report a case of idiopathic hypertrophic cranial pachymeningitis (IHCP) associated with an orbital pseudotumor and granulocytic thyroiditis of unknown origin. The patient suffered from progressive and finally complete loss of visual acuity, as well as from multiple cranial nerve dysfunctions. Medical treatment and radiation therapy were not beneficial. As reported in the literature, the CT and MRI findings of IHCP were non-specific; thus, diagnosis had to be made by exclusion and open biopsy was required. An inflammatory pseudotumor of the orbit is rarely found in patients with IHCP. Thyroiditis has not yet been reported in the context of IHCP. The patient also suffered from anosmia, which is an unusual feature of IHCP. PMID- 10460383 TI - Presacral extramedullary haematopoiesis with involvement of the sciatic nerve. AB - We present the case of a 60-year-old woman with no known blood disease who developed an extramedullary haematopoiesis of presacral localization that affected the right sciatic nerve. The diagnosis was made with imaging studies and CT-guided fine-needle aspiration. PMID- 10460384 TI - Abdominal retained surgical sponges: CT appearance. AB - Retention of surgical sponges is rare. They cause either an aseptic reaction without significant symptoms or an exudative reaction which results in early but nonspecific symptoms. Computed tomography is very useful for recognition of retained sponges. The appearance of retained sponges is widely variable. Air trapping into a surgical sponge results in the spongiform pattern which is characteristic but unfortunately uncommon. A low-density, high-density, or complex mass is found in the majority of cases, but these patterns are not specific. Sometimes, a thin high-density capsule may be seen. Rim or internal calcification is a rare finding. Finally, a radiopaque marker is not a reliable sign. Differentiation from abscess and hematoma is sometimes difficult. PMID- 10460385 TI - MR cholangiopancreatography: prospective comparison of a breath-hold 2D projection technique with diagnostic ERCP. AB - The aim of this study was to compare prospectively a breath-hold projection magnetic resonance cholangiopancreatography (MRCP) technique with diagnostic endoscopic retrograde cholangiopancreatography (ERCP). Seventy-six patients with suspected strictures or choledocholithiasis were referred for MRCP and subsequent ERCP examination, which were performed within 4 h of each other. The MRCP technique was performed using fat-suppressed rapid acquisition with relaxation enhancement (RARE) projection images obtained in standardised planes with additional targeted projections as required by the supervising radiologist. Two radiologists (in consensus) assessed the MRCP results prospectively and independently for the presence of bile duct calculi, strictures, non-specific biliary dilatation and pancreatic duct dilatation, and recorded a single primary diagnosis. The ERCP was assessed prospectively and independently by a single endoscopist and used as a gold standard for comparison with MRCP. Diagnostic agreement was assessed by the Kappa statistic. The MRCP technique failed in two patients and ERCP in five. In the remaining 69 referrals ERCP demonstrated normal findings in 23 cases, strictures in 19 cases, choledocholithiasis in 9 cases, non specific biliary dilatation in 14 cases and chronic pancreatitis in 4 cases. The MRCP technique correctly demonstrated 22 of 23 normal cases, 19 strictures with one false positive (sensitivity 100 %, specificity 98 %), all 9 cases of choledocholithiasis with two false positives (sensitivity 100 %, specificity 97 %), 12 of 14 cases of non-specific biliary dilatation and only 1 of 4 cases of chronic pancreatitis. There was overall good agreement for diagnosis based on a kappa value of 0.88. Breath-hold projection MRCP can provide non-invasively comparable diagnostic information to diagnostic ERCP for suspected choledocholithiasis and biliary strictures and may allow more selective use of therapeutic ERCP. PMID- 10460386 TI - Palliation of malignant esophageal strictures: initial results with self expanding uncovered nitinol coil stents. AB - The purpose of this study was to determine the efficacy of the uncovered coil stents in patients with malignant dysphagia. Coiled spring-shaped uncovered self expanding metallic Esophacoil stents (Instent, Eden Prairie, Minnesota) were placed in 11 patients (9 men and 2 women; age range 38-77 years, mean age 60.5 years) with malignant esophageal strictures and dysphagia, under fluoroscopic guidance. Dysphagia was graded on a scale of 0 to 4 (0 = no dysphagia; 1 = dysphagia to normal solids; 2 = dysphagia to soft solids; 3 = dysphagia to solids and liquids; 4 = complete dysphagia, inability to swallow saliva). Two patients had received radiation therapy, 4 had had chemotherapy, and 5 had had a combination of both radiation and chemotherapy before stent palliation. Control clinical examinations and endoscopic or barium swallow studies were performed every 4 weeks until the patient died. The stents were well tolerated by all patients and were effective in 9 of 11 patients with malignant dysphagia. Complications of the procedure included incomplete opening of the stent in 1 case, migration in 1 case, transient pain in 8 cases, reflux in 3 cases and minor gastrointestinal bleeding in 2 cases. Stent migration in 1 case resulted in surgical intervention and incomplete opening of the stent allowed only partial improvement of dysphagia in 1 case. The quality of life significantly improved in all other patients. Mean survival time of the patients was 73 days (range 34-125 days) and no significant tumor ingrowth was detected during the follow-up period. Insertion of an Esophacoil has a good palliative effect on dysphagia in patients with malignant esophageal strictures with few complications. Although the stent is uncovered, tumor ingrowth and overgrowth were not observed in our study, possibly because of previous treatments. PMID- 10460387 TI - Sensitivity of transabdominal ultrasonography in detection of intraperitoneal fluid in humans. AB - The sensitivity and specificity of ultrasonography in detection of free intraperitoneal fluid is over 90 %. The lowest detectable volume of free fluid in humans is unknown. The distribution of intraperitoneal fluid was studied in 86 patients by transabdominal US in group A (n = 21, 10 ml of fluid), in group B (n = 15, 50 ml of fluid) and group C (n = 50, splenic trauma). Ultrasound detected fluid in 15 of 21 patients in group A, and in all patients in groups B and C. In group A 10 ml of fluid was found in 71 % of cases behind the bladder, and in only 5-14 % of cases in the upper abdomen. In group B 50 ml of fluid was found in all patients in the lower pelvis, but in only 20 % in Morison's pouch and in 7 % around the spleen. In group C 200-4500 ml of fluid was detected by US in 72 % of patients in the perisplenic space, in 60 % in Morison's pouch and in 42 % in the retrovesical space. Small volumes of free intraperitoneal fluid (10-50 ml) can be detected with current US scanners, but only near the site of injury. These results support the role of US as a primary imaging modality in abdominal trauma. PMID- 10460388 TI - Perforated jejunal diverticulitis as a rare cause of acute abdomen. AB - Jejunal diverticula is rare and in most cases without any symptoms. They become clinically relevant when complications, such as diverticulitis, malabsorption caused by bacterial overgrowth, intestinal hemorrhage, or obstruction, occur. In this case report a case of perforated jejunal diverticulitis is presented and the problems in finding the correct diagnosis are discussed. PMID- 10460389 TI - Intestinal stenosis from mesenteric injury after blunt abdominal trauma. AB - We report a case in which blunt abdominal trauma resulted in injury to the mesentery with subsequent ischemic stricture of the adjacent small bowel. We present CT images at the time of trauma and 5 weeks later when clinical signs of intestinal obstruction occurred. We include images of enteroclysis and angiography of this uncommon sequela of blunt abdominal trauma. At surgery, a stenotic small bowel loop was found adjacent to a healed defect in the mesentery. Histological examination of the resected segment showed mucosal and submucosal ischemia with mucosal ulceration, mural inflammation, and fibrosis. Posttraumatic intestinal stenosis subsequent to a mesenteric tear should be included in the differential diagnosis in a patient with a history of blunt abdominal trauma and signs of intestinal obstruction. PMID- 10460390 TI - Colosalpingeal fistula: a rare complication of colonic diverticular disease. AB - Diverticular disease is a common condition in Western countries. The formation of inflammatory fistulae, usually from sigmoid colon to bladder or vagina, can be a feature of complicated cases of the disorder and is normally an indication for surgical intervention. We present a case of colosalpingeal fistulation occurring secondary to diverticulitis, a complication which, to our knowledge, has not been previously reported in the radiological literature. As in this instance, the initial clinical presentation of this problem can often be non-specific, with localising symptoms occurring later. In our case, barium enema examination allowed good demonstration of the fistulous communication before the more specific symptoms were clinically apparent. PMID- 10460391 TI - Transcatheter embolization of hepatic arteriovenous fistulas in Rendu-Osler-Weber disease: a case report and review of the literature. AB - A patient with hereditary hemorrhagic telangiectasia and diffuse intrahepatic arteriovenous fistulas developed secondary high-output ventricular failure and pulmonary hypertension. A serial staged hepatic arterial coil embolization was performed with long-term resultant haemodynamic and clinical improvement. The methods of this procedure and related complications are discussed. PMID- 10460392 TI - Aortocaval fistula complicating abdominal aortic aneurysm: diagnosis with gadolinium-enhanced three-dimensional MR angiography. AB - With approximately 150 reported cases, fistulas between the abdominal aorta and inferior vena cava are rare. Preoperative clinical diagnosis of aortocaval fistula is difficult because the classical triad of abdominal pain, pulsatile abdominal mass, and abdominal machinery-like bruit may be absent in up to 50 % of patients. We report a case of aortocaval fistula complicating abdominal aortic aneurysm which was diagnosed preoperatively using breath-hold gadolinium-enhanced three-dimensional MR angiography. PMID- 10460393 TI - Pseudoaneurysm of the internal carotid artery after shrapnel injury in World War II: demonstration by CT angiography with 3D MIP reconstruction. AB - A case of pseudoaneurysm of the left internal carotid artery (ICA) after shrapnel injury is demonstrated by intra-arterial digital subtraction angiography (DSA) and computed tomography angiography (CTA) with subtraction technique. Although the pseudoaneurysm was well demonstrated by intra-arterial DSA, CTA was the only modality to demonstrate the three-dimensional shape of the perfused part of pseudoaneurysm and the aneurysmal neck, which affected the therapeutic strategy. The CTA technique is useful in the assessment of large pseudoaneurysms and for therapeutic planning. PMID- 10460394 TI - Multivariate analysis of the adjustment of the colour duplex unit for the differential diagnosis of lymph node alterations. AB - Intranodal angioarchitecture was used as criterion for the differential diagnosis of lymph node alterations by colour duplex sonography. The influence of the adjustment of the duplex unit on the diagnostic accuracy and on the rate of lymph nodes with detectable intranodal flow signals was tested. A total of 243 superficial lymph nodes in 127 patients were included in a retrospective study. Diagnoses were approved by histopathology (n = 173) or clinical follow-ups (n = 70 reactive lymph nodes). The intranodal angioarchitecture and sonomorphology were assessed. We used nonparametric discriminant analysis in order to define appropriate settings for transducer frequency, flow mode, power and threshold which result in the best diagnostic quality. Of the lymph nodes, 87 % displayed flow signals using a threshold of >/= 13 and the flow modes low or medium. Vascularisation was detected only in 47 % of the nodes assessed with different adjustments. Diagnostic accuracy dropped from 94 to 68-84 % if threshold was < 13 or power was less than -11 dB. An insufficient adjustment of the colour duplex unit decreases the chance of detecting intranodal flow signals and impairs the diagnostic quality of colour duplex sonography. Flow mode, power and threshold are the most important parameters which have to be adjusted properly. PMID- 10460395 TI - Speech processing in radiology. AB - The goal of this study was to evaluate the recognition rate, learning potential and amount of time needed to complete a report with the Philips speech recognition system SP 6000 (Philips, Best, The Netherlands). Four radiologists dictated reports of interventional radiology, MRI examinations of the musculoskeletal system and CT examinations of the thorax and abdomen with the Philips system using the German language. The recognition rate of each report and improvement rate after each learning phase of the Philips system was assessed. The time needed to complete a report using the Philips system was then compared with the time needed to complete a report using the tape-based system via a time analysis. The average recognition rate for the four radiologists using the Philips system was 79.6 %, which improved to 92.5 % after the third adaptation. Initially, the average time demand to dictate and correct one report was approximately 16.8 min, but this time decreased to 8.1 min after the third adaptation. In contrast, only 3. 6 min were needed to dictate and correct one report using the tape-based system. However, with the speech recognition system, dictation, correction and transcription of the report can be completed within 15 min, whereas with the tape-based system, it takes nearly 1 day. With the Philips system, speech recognition can reach as high as 95 % since each adaptation of the system improves the recognition rate by approximately 5 %. While the Philips system is associated with longer dictation times than the tape-based system, turn around time for a complete report is substantially shorter with the Philips system than the tape-based system. PMID- 10460396 TI - The number and distribution of computerised tomography scanners in Turkey. AB - The goal of this study was to investigate the number and distribution of CT scanners in Turkey. Our results show 173 CT scanners in Turkey in 1994, which equals 2.9 scanners per million people. All of the scanners are located in 45 cities, where 81 % of the population resides. The other 31 cities in Turkey have no scanners. Of the 173 scanners, 103 (59.6 %) are owned by the private sector and the other 70 are owned by the public sector. Of Turkey's CT scanners, 49.2 % are located in private health centres, 21.9 % in university hospitals, 16.7 % in Ministry of Health (MOH) hospitals, 10.4 % in private hospitals and 1.8 % in social security hospitals. PMID- 10460397 TI - Patient satisfaction and quality of care at four diagnostic imaging procedures: mammography, double-contrast barium enema, abdominal ultrasonography and vaginal ultrasonography. AB - The objective of this study was to measure patient satisfaction and to investigate the practical implications of monitoring the quality of care at four radiology procedures. A survey was conducted immediately after the examinations in eight radiology departments: 550 patients attending for mammography, 110 for double-contrast barium enema (DCBE), 97 for abdominal ultrasonography and 90 for vaginal ultrasonography. Outcome measures were seven questionnaire scales: pain, emotional distress, information received, staff's punctuality and technical ability, facilities, and general satisfaction. Response rate was 87 %. Multivariate regression analysis showed significant differences between procedures on all scales (p < 0.001). Differences considered to be of practical importance, i. e. >/= 7 scale points, were detected on five of the scales. Mammography and DCBE caused the most pain, and vaginal US and DCBE caused the most distress. The US procedures entailed dissatisfaction with information about the procedures. The DCBE patients recorded dissatisfaction with the staff's lack of punctuality, and these and the mammography patients recorded dissatisfaction with the facilities. The findings indicate a potential for improving patients' experiences. Several aspects of care, i. e. pain management, attention to the patient's emotional concerns, explanation of procedures, punctuality and quality of the facilities, can be improved. PMID- 10460399 TI - Quality of life research in urology. PMID- 10460398 TI - Quiz case of the month. Diagnosis: duodenal gangliocytic paraganglioma. PMID- 10460400 TI - Health-related quality-of-life studies in urology: conceptual and methodological considerations. AB - This paper provides an overview of the scientific and practical aspects of assessing health-related quality of life (HRQL). The paper provides a brief overview of developments in the field and then discusses the definition of HRQL. The different types of HRQL measure available to researchers are described, with specific references to urology measures. The paper then focuses on the conceptual and methodological difficulties inherent in patient-centred measures and describes the various means of ensuring scientific soundness in such measures. It concludes that HRQL measures offer much potential to those interested in patient outcomes but that their widespread introduction into research and practice must be carried out with caution. PMID- 10460401 TI - Quality-of-life aspects in urology - benign prostatic hyperplasia. PMID- 10460402 TI - Examining health-related quality of life in men treated for prostate cancer. PMID- 10460403 TI - When the bladder is gone: quality of life following different types of urinary diversion. AB - Methods aiming for continence after radical cystectomy for bladder cancer, e.g., continent cutaneous urinary diversion and orthotopic bladder substitution, have become clinically accepted alternatives to ileal conduit diversion in the past decade. The a priori assumption has been that improvement in the postoperative quality of life would be greater following the new methods than after ileal conduit diversion. Studies on the validity of this assumption have been hampered by a lack of consensus on what should be measured and how this should be done: in no two studies have identical test instruments been used. The frequent inclusion of patients with nonmalignant disease has also impeded comparative evaluation of reports. By and large, published studies do not seem to confirm that the new types of urinary tract reconstruction improve the quality of life of bladder cancer patients. Common for all methods are psychosocial and sexual problems. PMID- 10460404 TI - Quality-of-life assessment in bladder cancer. PMID- 10460405 TI - The impact of urinary incontinence on the quality of life of women. PMID- 10460406 TI - Quality-of-life issues in the treatment of testicular cancer. AB - Quality of life (QOL) issues in testis cancer have recently assumed great importance for both physicians and patients. Since most of the patients are going to be long-term survivors, with modern therapeutic approaches, psychosocial difficulties and sexual life problems may become one of the major long-term complications of testis cancer treatment. QOL studies available demonstrate that approximately 10% of the patients will suffer from enduring long-term psychological problems, namely anxiety, depression, fatigue, and disrupted intimate relationships. Since these problems develop unrelated to the therapeutic approach, one has to develop risk profiles predicting psychological illness, such as with psychological counseling, prior to the initiation of the therapy. Impairment of sexual life and infertility distress represent other long-term sequelae of testis cancer treatment. The highest incidence of sexual dysfunction develops within the first 6 months following therapy, with most patients recovering within the next 3 years, resulting in a 15% rate of long-term sexual dysfunction. This relatively high frequency of sexual problems warrants an adequate counseling before and after therapy. Future perspectives of QOL research in testis cancer has to concentrate on the development of a site- specific questionnaire. Since the different therapeutic strategies in clinical stage 1 testis cancer result in the same high cure rates but may encounter various levels of psychosocial distress, QOL appears to represent the most important endpoint end of different treatment modalities in the clinical setting of different treatment modalities and QOL documentation must be integrated in all clinical study protocolls. QOL studies are important issues in the evaluation of each new future method of treatment modality going to be established for testis cancer. PMID- 10460408 TI - Quality-of-life studies in patients with ambiguous genitalia. AB - No published study to date has adequately addressed the quality of life (QOL) of intersexuals, although three studies reviewed some of these aspects. A tool specifically for studies in intersexuality is being developed. A pilot survey tested by this author using questions from a population-specific, standardized QOL survey tool was evaluated by use on a small sampling of an intersexual patient test population. The group comprised ten adult intersexuals with an average age of 34.2 years (range: 22-47 years). Survey results disclosed that all considered themselves healthy and physically active. Family relationships were described as generally good, though in discussion most showed some family problems that centered on gender issues. Six had many friends, three had some, and one had few. Eight of the ten had experienced orgasm. Only two felt they trusted doctors. All but one were partially or completely satisfied with their overall physical appearance. Those who had surgery were distressed by the appearance of their genitals, some expressing complete dissatisfaction. With development of sensitive survey tools reflecting the attitudes and concerns of this special population, important issues relating to intersexuality can be addressed. PMID- 10460407 TI - Mental health, psychosocial functioning, and quality of life in patients with bladder exstrophy and epispadias - an overview. AB - Although there has been only limited clinical research on mental or psychosocial implications in patients with bladder exstrophy and epispadias, questions have been raised as to whether their life is of such questionable quality that a termination of pregnancy should be considered. A systematic overview of outcome studies published over the past three decades was carried out. In all, 1208 abstracts and 52 papers were read; only 10 (0.8%) papers focused on the mental or psychosocial outcome, but with diverse findings. However, most of the studies suffered from serious methodological deficiencies. Physical, mental, and psychosocial problems revealed in studies with reliable and valid instruments have clinical implications and underline the need for the further development of surgical and psychosocial interventions. Multicenter studies with a multimodal, prospective, and longitudinal design, based on semistructured interviews and specific questionnaires related to the disorder, are appropriate. PMID- 10460409 TI - What makes homologous chromosomes find each other in meiosis? A review and an hypothesis. AB - The conditions re reviewed that must be met by any model of long distance attraction and transport of homologous chromosomes to the points of intimate DNA synapsis. A proposal for possible mechanisms is presented. It includes transcription and repair factors acting on coding sequences as a preparatory step toward pairing, and the attachment of specific pairing proteins to these sequences. Double-strand break formation is prepared but not immediately completed at the same sites. It is concluded that DNA-DNA interactions cannot bridge the distances between homologous chromosomes in the nucleus, and it is suggested that protein chains are formed between homologous segments. These attach to homologous chains emanating from homologous sequences in other chromosomes, and the chains move along each other until the homologous DNA sequences meet. Then, if required, a synaptonemal complex is formed, and exchange can take place. PMID- 10460410 TI - Localization and phosphorylation of HP1 proteins during the cell cycle in mammalian cells. AB - Mammalian heterochromatin proteins 1 (HP1alpha, HP1beta, and HP1gamma) are nonhistone proteins that interact in vitro with a set of proteins that play a role in chromatin silencing, transcription, and chromatin remodeling. Using antibodies specific for each HP1 isoform, we showed that they segregate in distinct nuclear domains of human HeLa cells. By contrast, in mouse 3T3 interphase cells, HP1alpha and HP1beta are strictly colocalized. In mitotic HeLa cells, all of HP1alpha and a fraction of HP1beta and HP1gamma remain associated with chromosomes. Immunostaining of spread HeLa chromosomes showed that HP1alpha is mainly localized on centromeres as shown previously for HP1beta, while HP1gamma is distributed on discrete sites on the arms of chromosomes. Biochemical analysis showed that HP1alpha and HP1gamma are phosphorylated throughout the cell cycle, although more extensively in mitosis than in interphase, while HP1beta apparently remains unphosphorylated. Therefore, despite their extensive sequence conservation, mammalian HP1 isoforms differ widely in their nuclear localization, mitotic distribution and cell cycle-related phosphorylation. Thus, subtle differences in primary sequence and in posttranslational modifications may promote their targeting at different chromatin sites, generating pleiotropic effects. PMID- 10460411 TI - RADHA--a new male germ line-specific chromosomal protein of Drosophila. AB - A new chromosomal protein - RADHA - of Drosophila is described that is specific for the male germ line. It is encoded by a single-copy gene, located in the region 96C-D of D. melanogaster polytene chromosomes. Transcription of the radha gene is restricted to the primary spermatocyte stage. The protein initially accumulates in some of the Y-chromosomal lampbrush loops. After meiosis it is found in the nuclei of spermatids and might be involved in chromatin rearrangement processes in the male germ line. RADHA is a basic protein with a C terminal leucine zipper region and several segments capable of forming coiled coil structures. PMID- 10460412 TI - Identification and characterization of MmORC4 and MmORC5, two subunits of the mouse origin of replication recognition complex. AB - Two new members of the mouse origin recognition complex (ORC) have been cloned that are closely related to Saccharomyces cerevisiae ORC4 and ORC5 as well as to their human homolog. Both MmORC4p and MmORC5p have a putative nucleotide triphosphate binding motif. Transcription of MmORC4 and MmORC5 is not suppressed in mouse NIH3T3 fibroblasts made quiescent by serum starvation. The transcription levels of both ORC genes are constantly high in all phases of the cell cycle. A screen based on the two-hybrid approach suggests that the product of the ORC4 gene interacts with the ORC2, but not with the ORC1 protein. The conservation of structure among members of the ORC4- and ORC5-related family of proteins suggests that these proteins play a key role in the initiation of DNA replication in all eukaryotes. PMID- 10460413 TI - Microdissection and microcloning of rye (Secale cereale L.) chromosome 1R. AB - Chromosome 1R was microdissected and collected from mitotic metaphase spreads of rye (Secale cereale L.) by using glass needles. The isolated chromosomes were amplified in vitro by Sau3A linker adaptor-mediated polymerase chain reaction (PCR). After amplification, the presence of rye-specific DNA was verified by Southern hybridization. The second-round PCR products from five 1R chromosomes were cloned into a plasmid vector to create a chromosome-specific library, which produced approximately 220,000 recombinant clones. Characterization of the microclone library showed that the 172 clones evaluated ranged in size from 300 1800 bp with an average size of 950 bp, of which approximately 42% were medium/high copy and 58% were low/unique copy clones. Chromosome in situ hybridization confirmed that the PCR products from microdissected chromosomes originated from chromosome 1R, indicating that many chromosome 1R-specific sequences were present in the library. PMID- 10460414 TI - Chromosome engineering: generation of mono- and dicentric isochromosomes in a somatic cell hybrid system. AB - The most common isochromosome found in humans involves the long arm of the X, i(Xq), and is associated with a subset of Turner syndrome cases. To study the formation and behavior of isochromosomes in a more tractable experimental system, we have developed a somatic cell hybrid model system that allows for the selection of mono- or dicentric isochromosomes involving the short arm of the X, i(Xp). Simultaneous positive and negative counterselection of a mouse/human somatic cell hybrid containing a human X chromosome, selecting for retention of the UBE1 locus in Xp but against the HPRT locus in Xq, results in a variety of abnormalities of the X chromosome involving deletions of Xq. We have generated 70 such "Pushmi-Pullyu" hybrids derived from seven independent X chromosomes. Cytogenetic analysis of these hybrids using fluorescence in situ hybridization showed i(Xp) chromosomes in approximately 19% of the hybrids. Southern blot and polymerase chain reaction analyses of the Pushmi-Pullyu hybrids revealed a distribution of breakpoints along Xq. The distance between the centromeres of the dicentric i(Xp)s generated ranged from approximately 2 Mb to approximately 20 Mb. To examine centromeric activity in these dicentric i(Xp)s, we used indirect immunofluorescence with antibodies to centromere protein E (CENP-E). CENP-E was detected at only one of the centromeres of a dicentric i(Xp) with approximately 2 3 Mb of Xq DNA. In contrast, CENP-E was detected at both centromeres of a dicentric i(Xp) with approximately 14 Mb of Xq DNA. Two other dicentric i(Xp) chromosomes were heterogeneous with respect to centromeric activity, suggesting that centromeric activity and chromosome stability of dicentric chromosomes may be more complicated than previously thought. The Pushmi-Pullyu model system presented in this study may provide a tool for examining the structure and function of mammalian centromeres. PMID- 10460415 TI - Chromosome painting of Y chromosomes and isolation of a Y chromosome-specific repetitive sequence in the dioecious plant Rumex acetosa. AB - The dioecious plant Rumex acetosa has a multiple sex chromosome system: XX in female and XY(1)Y(2) in male. Both types of Y chromosome were isolated from chromosome spreads of males by manual microdissection, and their chromosomal DNA was amplified using degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR). When the biotin-labeled DOP-PCR product was hybridized with competitor DNA in situ, the fluorescent signal painted the Y chromosomes. A library of Y chromosome DNA was constructed from the DOP-PCR product and screened for DNA sequences specific to the Y chromosome. One Y chromosome-specific DNA sequence was identified and designated RAYSI (R. acetosa Y chromosome-specific sequence I). RAYSI is a tandemly arranged repetitive DNA sequence that maps to the 4',6 diamidino-2-phenylindole bands of both Y chromosomes. PMID- 10460416 TI - Vitreous humor fructosamine concentrations in the autopsy diagnosis of diabetes mellitus. AB - In clinical practice, biochemical markers, particularly serum glucose levels are used to diagnose diabetes mellitus. However, at autopsy this marker is of no value due to the substantial and capricious fluctuations in glucose levels after death. The aim of this study was to evaluate the usefulness of the postmortem determination of fructosamine in vitreous humor for confirming the presence of antemortem hyperglycemia. This was a study of 92 cadavers with a mean age of 60.05 years (SD 17.73) and a mean postmortem interval of 17.02 h (SD 9.76, range 2-58 h). Cases were assigned to two diagnostic groups according to the antemortem diagnosis of diabetes mellitus based on the patients' medical records. In vitreous humor statistically significant differences were found in glucose and fructosamine concentrations between the two diagnostic groups, the highest values being obtained in the group of subjects with a previous diagnosis of diabetes mellitus. PMID- 10460417 TI - Histological demonstration of haemosiderin deposits in lungs and liver from victims of chronic physical child abuse. AB - In the context of chronic physical child abuse, two entities have been described based on macroscopical and radiological criteria: the battered baby syndrome and the shaken baby syndrome. However, in some autopsy cases, clinico-radiological information may not be available. In these cases, histological examinations are necessary to look for sequelae of repeated haemorrhages, particularly in organs likely to have suffered traumatisms such as the lungs, or in organs belonging to the mononucleated macrophage resorption system, such as the liver and the spleen. We examined a series of 15 young children who died from proven chronic child abuse and compared them with 15 sex and age-matched control subjects who died from natural causes with no history of child abuse. Using Perl's stain for iron, we identified haemosiderin deposits in pulmonary, hepatic and splenic samples and the deposits were evaluated qualitatively and quantitatively. Haemosiderin deposits were significantly (P < 0.001) more abundant in the lungs and liver of the chronic abuse victims than in those of the control subjects. However, they were not significantly more abundant in the spleens of child abuse victims than in controls. We conclude that haemosiderin deposits in lungs and liver could be proposed as a marker for chronic physical child abuse. This study stresses the importance of systematic histological examination to look for pulmonary and hepatic haemosiderin deposits in cases in which chronic child abuse is suspected. PMID- 10460418 TI - Mitochondrial DNA typing from human axillary, pubic and head hair shafts - success rates and sequence comparisons. AB - The analysis of mitochondrial DNA (mtDNA) from shed hairs has gained high importance in forensic casework since telogen hairs are one of the most common types of evidence left at the crime scene. In this systematic study of hair shafts from 20 individuals, the correlation of mtDNA recovery with hair morphology (length, diameter, volume, colour), with sex, and with body localisation (head, armpit, pubis) was investigated. The highest average success rate of hypervariable region 1 (HV 1) sequencing was found in head hair shafts (75%) followed by pubic (66%) and axillary hair shafts (52%). No statistically significant correlation between morphological parameters or sex and the success rate of sequencing was found. MtDNA sequences of buccal cells, head, pubic and axillary hair shafts did not show intraindividual differences. Heteroplasmic base positions were observed neither in the hair shafts nor in control samples of buccal cells. PMID- 10460419 TI - Expanding the forensic German mitochondrial DNA control region database: genetic diversity as a function of sample size and microgeography. AB - Mitochondrial DNA control region sequences were determined in 109 unrelated German Caucasoid individuals from north west Germany for both hypervariable regions 1 (HV1) and 2 (HV2) and 100 polymorphic nucleotide positions (nps) were found, 63 in HV1 and 37 in HV2. A total of 100 different mtDNA lineages was revealed, of which 7 were shared by 2 individuals and 1 by 3 individuals. The probability of drawing a HV1 sequence match within the north west Germans or within published sets of south Germans and west Austrians is similar (within a factor of 2) to drawing a sequence match between any two of these three population samples. Furthermore, HV1 sequences of 700 male inhabitants of one village in Lower Saxony were generated and these showed a nearly linear increase of the number of different haplotypes with increasing number of individuals, demonstrating that the commonly used haplotype diversity measure (Nei 1987) for population samples tends to underestimate mtDNA diversity in the actual population. PMID- 10460420 TI - Fatal poisonings where ethylmorphine from antitussive medications contributed to death. AB - The hypothesis that antitussives containing ethylmorphine are abused by alcoholics and drug addicts and that this may lead to fatal poisonings where ethylmorphine causes or contributes to death was investigated. For this purpose 14 cases were analysed where a blood ethylmorphine concentration above the therapeutic level of >/= 0.3 microg/g was found in autopsy blood samples. Alcohol was found in 8 of the 14 cases and alcoholism or drug addiction was noted on 8 of the 14 death certificates. Other drugs, mostly benzodiazepines, were found in all 14 cases. The cause of death was fatal poisoning in 8 of the 14 cases and although there were no mono-intoxications, the cause of death was specified as fatal ethylmorphine poisoning in 2 cases. Among the unspecified medicinal drug poisonings there were five cases with very high blood levels of ethylmorphine, indicating that this drug played an important contribution to the cause of death. The results indicate that deaths due to ethylmorphine in antitussive medicines may occur among drug addicts and alcoholics taking it in overdose. Physicians should therefore be restrictive in prescribing cough mixtures containing ethylmorphine to these categories of patients. Prescription of large amounts of the drug should be avoided. PMID- 10460421 TI - Iatrogenic staphylococcus aureus septicaemia following intravenous and intramuscular injections: clinical course and pathomorphological findings. AB - The clinical course, autopsy and histological findings are presented from three (one 33-year-old female and two males aged 26 and 56) fatalities resulting from injection therapy which has produced Staphylococcus aureus septicaemia. The autopsies were performed within 2-4 days postmortem. No primary focus other than the insertion site of the peripheral venous catheters or the intramuscular injections, representing the initial entry site of Staphylococcus aureus, could be identified. Death was attributed directly to the staphylococcal infection as a result of iatrogenic injection therapy for the treatment of a non-severe underlying illness (premature labour pains, acute loss of hearing, lumbago). The forensic diagnosis of Staphylococcus aureus septicaemia following iatrogenic injections has to be critically evaluated and can be established routinely in cases with delayed autopsy only when no other cause of death is revealed by autopsy, no apparent source of infection other than the insertion site can be detected and careful attention is paid to histological and bacteriological findings. All doubtful cases of nosocomial bloodstream infections with fatal outcome should undergo an immediate autopsy. In cases of very early forensic involvement microbiological investigations, including phagotyping, molecular biological characterization and identification of bacterial toxins from micro organisms out of appropriate specimens obtained postmortem, could be efforts of potential evidential value regarding the aetiological proof. To optimize aetiopathogenetic conclusions concerning a causal relationship between iatrogenic injections and septic complications, the medicolegal investigation should also include an interdisciplinary co-operation with consultants from other relevant fields (e.g. microbiology and hygienics). PMID- 10460422 TI - Munchausen syndrome by proxy and intra-alveolar haemosiderin. AB - Munchausen syndrome by proxy is characterised by the invention of a false history and/or the deliberate inducement of a factitious illness by parents in their child. First reported as a disorder of mothers, this syndrome is now recognised to have male perpetrators. One of the most common characteristic presentations is with the child allegedly suffering repeated apnoeic attacks. These are in fact deliberately induced episodes of upper airway obstruction. The children of these parents are at great risk of serious harm or death if not recognised. The identification of smothering at autopsy is fraught with difficulty and the presence of intra-alveolar haemosiderin has been claimed to be a marker of previous smothering. A case of Munchausen syndrome by proxy is presented where there were deliberate acts of repeated partial smothering and where the finding of intra-alveolar haemosiderin at autopsy provided additional supportive evidence of smothering. PMID- 10460423 TI - Fatal brain injury caused by the free-flying blade of a knife - case report and evaluation of the unusual weapon. AB - A man suffered a fatal injury from a self-inflicted accident while handling a special type of knife. A spring in the shaft of the knife accelerated the blade, which perforated the orbital cavity and the frontal lobe at the right side. Death was due to central disregulation. The initial velocity of the blade was measured to be 15 m/s. In a total of 20 experimental shots to a fresh pig cadaver, the blade always penetrated the skin and 5-10 cm of soft tissue as long as the distance did not exceed 1 m. Thin layers of bone were also perforated. The free flight of the blade did not remain stable if the distance was more than 1 m, which resulted in superficial wounds only. So this unusual construction resembling a knife can be considered an effective combat weapon for close range fighting instead of a tool. PMID- 10460424 TI - Forensic mtDNA hair analysis excludes a dog from having caused a traffic accident. AB - A dog was suspected of having caused a traffic accident. Three hair fragments were recovered from the damaged car and subjected to DNA sequence analysis of the canine mitochondrial D-loop control region. The results were compared to saliva and hair samples from the alleged dog, as well as to control hair samples from four unrelated dogs of different breeds. Two sequence types exhibiting five nucleotide differences in a 377 bp fragment were identified among the four controls. Whereas the evidence hair fragment was identical to the type 1 control sequence, the alleged dog shared the type 2 control sequence except for one position. Thus the dog could be excluded as the origin of the hair fragment. As canine mtDNA appears to exhibit only limited polymorphism, mitochondrial D-loop sequence comparison is currently only suitable for exclusions. PMID- 10460425 TI - Perimortem fixation of the gastric and duodenal mucosa: a diagnostic indication for oral poisoning. AB - Two cases of fatal oral poisoning are presented. In the first case, a 40-year-old man died due to a lethal dose of mercury (blood concentration 113.8 microg/ml) and in the second, a 34-year-old man died of chloralhydrate overdose with a lethal blood concentration of trichloroethanol (52 microg/ml), the active metabolite of chloralhydrate. In both cases gross examination and histology showed an unusually well preserved gastrointestinal mucosa in addition to unspecific signs of intoxication. The two cases demonstrate that the phenomenon of perimortal fixation is a useful indication for the forensic pathologist and should direct the suspicion to oral poisoning. The detection of fixation facilitates toxicology screening by indicating that the relevant substance must have the capability to precipitate proteins. PMID- 10460426 TI - Non-traumatic liver rupture due to a perforated gastric ulcer. AB - The case of a 57-year-old woman with a fatal liver rupture due to a necrotizing perihepatic abscess caused by a perforated gastric ulcer is presented. The ulcer had been treated successfully by surgical intervention 8 days before. The autopsy revealed a large perihepatic abscess and multiple ruptures of Glisson's capsule with a large subcapsular hematoma and underlying lacerations of the liver parenchyma. The patient had no history of previous abdominal trauma and the known etiological factors for spontaneous liver rupture were excluded by the autopsy findings or by clinical and laboratory data. No liver penetration by the gastric ulcer was found at autopsy and there were no clinical signs or symptoms for an infection or any degenerative or inflammatory diseases. Histologically abundant vegetable fibers, identified as stomach contents and a dense infiltrate of lymphocytes and granulocytes were found in the perihepatic abscess next to Glisson's capsule. Below Glisson's capsule there were hemorrhages, focal hepatocellular necrosis and a mixed cell inflammatory infiltration. In the present case, preceding perforation of the gastric ulcer with leaking of gastric acid into the peritoneal cavity resulted in peptic digestion of Glisson's capsule. Vascular lesions of the affected parts of Glisson's capsule and the liver parenchyma underneath resulted in intrahepatic hemorrhage and an increase in intrahepatic pressure with subsequent liver rupture. To the authors' knowledge no similar case of spontaneous liver rupture due to perforation of a gastric ulcer has been reported previously. PMID- 10460427 TI - German shepherd dog is suspected of sexually abusing a child. AB - A rare case of provoked anal penetration of an 11-year-old boy by a male German shepherd dog was confirmed by the results of morphological, serological and molecular genetic investigations. These results were of great importance to refute the suspicion on two adults. Some serious doubts remained in the version of the course of the event as presented by the boy. Some weeks later when confronted by a psychologist, the boy admitted having deliberately stimulated the dog manually and caused the animal to penetrate him. PMID- 10460428 TI - Allele frequencies of three STRs of the human von Willebrand factor gene (vWF) in a Brazilian population sample. AB - Allele frequencies were calculated for three tetrameric short tandem repeats (STRs) located in intron 40 of the human von Willebrand factor (vWA, vWF1 and vWF2) in 352 white individuals sampled from an urban population from the northeastern region of the State of Sao Paulo, Brazil. The exact test did not indicate any significant deviation from HWE for any of the three investigated loci. The allele frequencies of vWA and vWF1 showed unimodal and bimodal distributions, respectively, and the frequencies of vWF2 in our sample exhibited bimodal or even trimodal patterns. These differing patterns could reflect the differential action of one selective factor or of the distribution of mutations in these STRs, although the STRs are very close to one another and belong to the same gene. The frequency of paternity exclusions observed for each of these three loci conform to the theorectical expectations. The lack of difficulties regarding the methodology of typing and the forensic value of statistical parameters confirm the usefulness of these systems to study Brazilian populations. PMID- 10460429 TI - Manipulated radiographic material--capability and risk for the forensic consultant? AB - As interest is being increasingly focused on the digital processing of radiographs for identification of the deceased, the benefits and risks of electronic image processing are presented. With digitization of all kinds of radiographic equipment being on the increase and image processing personal computers being readily accessible, increasing quantities of manipulated radiographic material are to be expected in the future. This potential risk is meanwhile highlighted from the legal aspect. PMID- 10460430 TI - DNA typing in cases of blood chimerism. AB - A chimera is an organism whose cells derive from two or more distinct zygote lineages. and therefore two different blood cell populations circulate in one individual. To point out the potential pitfalls in forensic analysis, a set of triplets (a girl and two boys) who revealed blood chimerism was investigated with four STR systems using PCR. The results indicated that a DNA profile based on DNA extracted from blood can lead to a false determination because the band pattern of each triplet contained a mixture of the original genotype and the genotype of the siblings. Additional investigations on biological materials other than blood must be made in order to find out the real genetic characteristics of each child. PMID- 10460431 TI - Allele frequencies for the VNTR locus D17S5 (YNZ22) in Hungary. AB - A population genetic study for the locus D17S5 was carried out on Caucasoids from the Budapest area consisting of 209 unrelated individuals. In this system we identified 13 different alleles and 51 genotypes. No new alleles were found and no significant deviation from Hardy-Weinberg equilibrium was observed. The power of discrimination was estimated at 0.958, the power of exclusion at 0. 60 and the observed heterozygosity at 0.80. PMID- 10460432 TI - Genetic data on three complex STRs (ACTBP2, D21S11 and HUMFIBRA/FGA) in the Galician population (NW Spain). AB - The allele frequency distributions of three complex STRs, ACTBP2 (SE33), D21S11 and HUMFIBRA/ FGA in the population of Galicia were investigated. Analysis was carried out under denaturing conditions and fluorescent detection in the ALF DNA sequencer and typing was made by comparison with sequenced allelic ladders. No significant deviations from Hardy-Weinberg equilibrium were observed. No significant differences were found between our data and other Caucasian population data. ACTBP2 seems to be one of the most informative and polymorphic STRs. PMID- 10460433 TI - Spanish population data on the four STR loci D8S1179, D16S539, D18S51 and D21S11. AB - Population data were generated for four tetrameric short tandem repeat loci systems (D8S1179, D16S539, D18S51 and D21S11) for a Spanish Caucasian population sample (n = 218-219 individuals) using PCR. All loci were highly polymorphic, met Hardy-Weinberg expectations and the results demonstrated the assumption of independence of the loci analysed. The allele frequency data can be used in identity testing to estimate the frequency of a multiple PCR-based DNA profile in the Spanish population. PMID- 10460434 TI - Modified primers for D12S391 and a modified silver staining technique. AB - In this paper we describe a new primer pair for the short tandem repeat (STR) D12S391 which makes it possible to obtain considerably shorter amplification fragments (125-173 bp), compared to the previously published primers (205-253 bp). The primers were tested on 70 samples with known genotypes, and no differing results were found. In sensitivity studies, forensic casework samples and DNA quality studies, we proved that the new primers can improve the efficiency of the amplification. Moreover, the resolution of this locus on denaturing PAGE followed by silver staining was dramatically improved. This improvement was found to be most valuable for typing the rare.3 variants known for this locus. We also present and propose a new method for silver staining denaturing acrylamide gels. PMID- 10460435 TI - Allele distribution of three STRs in a population from Mozambique. AB - The short tandem repeat systems (STRs) D12S391, HUMFIBRA/FGA and HUMACTBP2 (SE33) were studied in a population of unrelated individuals from the Maputo area of south Mozambique. PMID- 10460436 TI - Correct mitochondrial L-strand sequencing after C-stretches. W. Parson et al. Int J Legal Med (1998) 111: 124-132. PMID- 10460438 TI - Transcranial color-coded duplex sonography. AB - Transcranial color-coded duplex sonography (TCCS) enables the reliable assessment of intracranial stenoses, occlusions, and cross-flow through the circle of Willis without using potentially hazardous compression tests. Transpulmonary ultrasound contrast agents (UCAs) increase the number of conclusive TCCS investigations, which suggests that UCAs may provide the conclusive evaluation of intracranial arteries in most patients with ischemic cerebrovascular disease. Further, contrast-enhanced TCCS may become an important tool both for the management of acute ischemic stroke by assessing intracranial hemodynamics and the displacement and diameter changes in supratentorial ventricles. TCCS is useful for the detection and monitoring of intracranial vasospasm, may visualize larger supratentorial hematomas with subcortical location and hemorrhagic transformation of ischemic infarcts, and provides the incidental detection of cerebral aneurysms and arteriovenous malformations. Second-generation UCAs and new ultrasound machines are very likely to further increase the frequency of conclusive TCCS studies. Power-based three-dimensional, contrast-enhanced TCCS is an important further development, which would make the method much less operator dependent. Site-targeted UCAs appear to provide a new and exciting method for ultrasonic diagnosis and management of patients with ischemic cerebrovascular disease. PMID- 10460439 TI - The stiff man and stiff man plus syndromes. AB - Rigidity in the setting of continuous motor unit activity at rest can be caused by a variety of central and peripheral conditions. A central origin is suggested by the presence of painful reflex spasms. Focal spinal lesions and infective causes are relatively easily excluded through imaging, microbiological and serological studies. There then remain a group of patients who may have the classical 'stiff-man syndrome' or a related syndrome. When strict diagnostic criteria are used, patients with the stiff man syndrome uniformly have axial rigidity, and about 90% are found to have antibodies against glutamic acid decarboxylase. Treatment response and prognosis are excellent. Stiff persons with 'plus' signs, particularly those with rigidity of a distal limb, are unlikely to have the classical stiff man syndrome. They have a poorer treatment response and prognosis. Some have a paraneoplastic aetiology, while a non-malignant autoimmune basis seems likely in others. Those in whom post-mortem pathology findings are available usually are seen to have had an encephalomyelitis with prominent involvement of the grey matter. Clinically, stiff persons with 'plus' signs may be divided into three groups according to the aggressiveness of the pathology and its relative distribution. Encephalomyelitis with rigidity follows a relentless subacute course, leading to death within 3 years. Chronic cases may present with predominantly brainstem involvement, including generalised myoclonus (the 'jerking stiff person syndrome') or spinal cord involvement, dominated by stiffness and spasm in one or more limbs (the 'stiff limb syndrome'). PMID- 10460440 TI - Chance and design. PMID- 10460441 TI - Leukoencephalopathy with cerebral amyloid angiopathy: a semiquantitative and morphometric study. AB - To investigate changes in caliber of vessels in leukoencephalopathy with cerebral amyloid angiopathy (CAA) we performed a histological and morphometric study of cerebral arteries in this disease. We histologically examined changes in cortico leptomeningeal arteries in five cases of leukoencephalopathy with CAA and compared their morphometrically determined wall-to-lumen ratio [(external diameter-internal diameter) x 0.5/internal diameter] with those of amyloid negative arteries to estimate stenotic changes. Additionally, we compared wall-to lumen ratios of medullary arteries in brains with CAA and white matter lesions (WML) (CAA(+)/WML(+), n = 5), subcortical arteriosclerotic encephalopathy without CAA (CAA(-)/WML(+), n = 7), and neither CAA nor white matter lesions (CAA(-)/WML( ), n = 5). Amyloid-positive arteries had thinned walls and dilated lumens. The external diameter and the wall-to-lumen ratio for amyloid-positive arteries was smaller than for amyloid-negative arteries in CAA(+)/WML(+) brains. There was no significant difference in the external diameters among the three groups. The wall to-lumen ratio for medullary arteries in CAA(-)/WML(+) brains was significantly greater than for CAA(+)/WML(+) and CAA(-)/WML(-), but there was no significant difference between CAA(+)/WML(+) and CAA(-)/WML(-). Amyloid deposition causes degeneration of the tunica media, resulting in thinning of the wall and dilation of the lumen. The tunica media of small arteries is important in regulation of cerebral blood flow with degeneration causing impairment of cerebrovascular autoregulation in response to blood pressure. This impairment may lead to white matter lesions. PMID- 10460442 TI - Plant-induced seizures: reappearance of an old problem. AB - Several plant-derived essential oils have been known for over a century to have epileptogenic properties. We report three healthy patients, two adults and one child, who suffered from an isolated generalized tonic-clonic seizure and a generalized tonic status, respectively, related to the absorption of several of these oils for therapeutic purposes. No other cause of epilepsy was found, and outcome was good in the two adult cases, but the course has been less favorable in the child. A survey of the literature shows essential oils of 11 plants to be powerful convulsants (eucalyptus, fennel, hyssop, pennyroyal, rosemary, sage, savin, tansy, thuja, turpentine, and wormwood) due to their content of highly reactive monoterpene ketones, such as camphor, pinocamphone, thujone, cineole, pulegone, sabinylacetate, and fenchone. Our three cases strongly support the concept of plant-related toxic seizure. Nowadays the wide use of these compounds in certain unconventional medicines makes this severe complication again possible. PMID- 10460443 TI - Limb contractures in levodopa-responsive parkinsonism: a clinical and investigational study of seven new cases. AB - We describe six patients with classical levodopa-responsive Parkinson's disease (PD) and one case of levodopa-responsive familial juvenile dystonia-parkinsonism with fixed contractures of the hands, feet or legs. In most patients contractures became established over a short period (2 months-2 years) but a considerable time after onset of parkinsonism (mean 13 years). Mean disease duration was 17 years, and all patients had severe levodopa-induced dyskinesias, either biphasic or peak dose, in the affected limb prior to onset of the contracture. Nerve conduction studies excluded peripheral ulnar nerve lesions in all patients with one exception, who was found to have a mild bilateral ulnar entrapment neuropathy. Transcranial magnetic stimulation performed in five of the seven patients showed shorter mean central motor conduction time in the affected than in the unaffected limb. Results of magnetic resonance imaging of the brain performed in a subgroup of patients were normal, with no evidence to suggest multiple system atrophy, cerebral infarction or focal abnormalities of the basal ganglia. We conclude that hand and feet contractures are not necessarily restricted to parkinson plus syndromes and may complicate otherwise typical PD in the absence of a structural or peripheral nervous cause. Striatal dopaminergic deficiency, particularly long standing, may have a role in the pathogenesis of limb contractures in PD. PMID- 10460444 TI - Dysphagia and nutritional status in multiple sclerosis. AB - In this observational study of patients with multiple sclerosis (MS) admitted to a regional neurology centre we assessed the frequency of dysphagia (objectively defined), dysphagia related symptoms, bulbar signs and nutritional status. We studied 79 consecutive admissions with MS (24 at diagnostic admission and 55 more advanced cases admitted for treatment and/or rehabilitation): normative swallowing data were from 181 healthy controls. Swallowing symptoms and signs were semi-quantitatively measured and compared to healthy controls. Dysphagia was defined by a quantitative water test. Disability was determined by Kurtzke's Expanded Disability Status Scale and Barthel's index. Nutritional status was assessed by body mass index, estimated percentage body fat from skin fold thickness measurements at four sites, a global evaluation of nutrition, the presence of pressure sores and the pressure sore risk using the Waterlow score. Patients with MS were more likely to complain of abnormal swallowing, of coughing when eating, and of food 'going down the wrong way' than healthy controls (P < 0.005). These significantly associated symptoms had high specificity but relatively low sensitivity. 43% of patients had abnormal swallowing, almost half of whom did not complain of it: abnormal swallowing was associated with several factors including abnormal brainstem/cerebellar function, disability, vital capacity, and depression score. Those with abnormal swallowing had higher Waterlow scores (P < 0.001), but, overall, abnormal swallowing was not associated with a difference in nutritional indices or incidence of pressure sores. In summary, abnormal swallowing is common in MS although often not complained of. It is associated with disordered brainstem/cerebellar function, overall disability, depressed mood and low vital capacity. It was not associated with major nutritional failure or pressure sores in this study. PMID- 10460445 TI - Stroke following chiropractic manipulation of the cervical spine. AB - We analyzed the clinical course and neuroradiological findings of ten patients aged 27-46 years, with ischemic stroke secondary to vertebral artery dissection (VAD; n = 8) or internal carotid artery dissection (CAD; n = 2), all following chiropractic manipulation of the cervical spine. The following observations were made: (a) All patients had uneventful medical histories, no or only mild vascular risk factors, and no predisposing vascular lesions. (b) VAD was unilateral in five patients and bilateral in three. VAD was located close to the atlantoaxial joint in all eight patients and showed additional involvement of lower sections in six, as well as temporary occlusion of one vertebral artery in three. (c) Nine of ten patients had brain infarction documented by magnetic resonance imaging or computed tomography. (d) Onset of symptoms was immediately after the manipulation (n = 5) or within 2 days (n = 5). (e) Progression of neurological deficits occurred within the following hours to a maximum of 3 weeks. (f) Maximum neurological deficits were severe in nine of ten patients. (g) Outcome after 4 weeks-3 years included no or mild neurological deficits in five patients, marked deficits in three, persistent locked-in syndrome in one, and persistent vegetative state in one. (h) Informed consent was obtained in only one of ten patients. Thus, patients at risk for stroke after chiropractic manipulation may not be identified a priori. Neurological deficits may be severely disabling and are potentially life threatening. PMID- 10460446 TI - Disease activity in multiple sclerosis studied by weekly triple-dose magnetic resonance imaging. AB - This study assessed whether dysfunction of the blood-brain barrier is an obligatory early event in lesion formation in multiple sclerosis. Dual-echo and T1-weighted magnetic resonance imaging after the injection of a triple dose (0.3 mmol/kg) of gadolinium-DTPA were obtained from ten patients with relapsing remitting multiple sclerosis every week for 2 months. Sixty-four newly active lesions were detected by the two techniques. All the 44 new lesions seen on dual echo scans enhanced during the early phases of their formation: 33 at their first appearance, 10 1 week before their appearance on the dual-echo scans, and one the week thereafter. When the every fourth (monthly) scan was analyzed, a total of 55 newly active lesions were detected (i.e., 14% active lesions would have been missed compared to the number found on weekly scanning). Thirty-one of them were detected by both dual-echo and triple-dose scans, 15 only by enhanced scans, and nine only by dual-echo scans. This study confirms that with highly sensitive magnetic resonance imaging techniques dysfunction of the blood-brain barrier is an obligatory early event in new lesion formation in relapsing-remitting multiple sclerosis. PMID- 10460447 TI - Magnetic resonance proton spectroscopy of the brain in Wilson's disease. AB - We studied 13 patients with Wilson's disease (WD) using localized magnetic resonance proton spectroscopy to test whether hepatic encephalopathy or impaired energy metabolism contributes to neurological dysfunction. Levels of myoinositol (MI), choline-containing compounds (Cho), creatine (Cr), N-acetyl-aspartate (NAA), Glx (unresolved resonances of glutamate, glutamine, and gamma-aminobutyric acid) and lactate were measured using a relative quantitative approach. Results were compared with those from 12 healthy controls. In one patient with de novo WD and acute hepatic disease but no neurological symptoms we found a marked decrease in the Cho/Cr and MI/Cr ratios. However, proton spectroscopy in the white matter, gray matter, and putamen of patients with treated WD showed no significant differences compared to healthy controls. In none of the subjects studied was the lactate/Cr ratio elevated. The spectroscopic findings were compatible with subclinical hepatic encephalopathy in the one patient with de novo WD and acute hepatic disease, but this does not play a major role in brain dysfunction in patients with treated WD. Additionally, there was no evidence of increased lactate concentration, indicating that cerebral energy metabolism was not grossly impaired. PMID- 10460448 TI - Outcome of central pontine and extrapontine myelinolysis (n = 44). AB - The findings in 44 patients (42 of whom were chronic alcoholics) with central pontine myelinolysis show that the outcome does not depend on the severity of neurological deficits during the acute phase of the condition or on concomitant internal diseases, including the degree of hyponatremia. Of the 34 patients for whom follow-up data were available, 32 survived. Of these 11 completely recovered, 11 had some deficits but were independent, and 10 were dependent (4 through disorders of memory or cognition, 3 with tetraparesis, 2 with cerebellar ataxia, 1 with polyneuropathy). The electrophysiological findings did not contribute usefully to the prediction of outcome. Additional neuroradiological diagnostic testing with magnetic resonance imaging was also of no prognostic significance. The extent of the initial pontine lesion was not correlated with the severity of clinical findings during the acute phase of disease, nor was persistence of the pontine lesion as usually seen on magnetic resonance imaging correlated with clinical improvement. We conclude that patients with cerebral myelinolysis survive if the nonspecific secondary complications of transient illnesses such as aspiration pneumonia, ascending urinary tract infection with subsequent septicemia, deep venous thrombosis, and pulmonary embolism can be avoided. PMID- 10460449 TI - Cerebral computed tomography and electroencephalography compared with neuropsychological findings in systemic lupus erythematosus. AB - Central nervous system involvement was evaluated in 36 patients with systemic lupus erythematosus (SLE) using cerebral computed tomography (CT), electroencephalography (EEG), and a neuropsychological test battery. The purpose was to investigate whether brain dysfunction as assessed by comprehensive neuropsychological investigation is associated with findings of routine investigation methods such as CT and EEG which are available in most hospitals. Abnormal EEG was found in 19%, and CT revealed cerebral atrophy in 47% of SLE patients. Few neuropsychological functions were affected by the presence of abnormal EEG, cerebral atrophy, or infarcts. Significant associations were found only between cortical atrophy and impairment of tactile spatial problem-solving and motor dexterity, and between cortical infarcts and motor dexterity in the dominant hand. The value of conventional EEG in assessing cerebral SLE is negligible, except for identifying epileptic activity and focal pathology. Cerebral CT has little relevance in predicting brain dysfunction as established by neuropsychological assessment in SLE, except for detecting cortical atrophy and infarcts. PMID- 10460450 TI - Dissections after childbirth. AB - The occurrence of spontaneous internal carotid or vertebral artery dissection after childbirth remains rare. To our knowledge, seven cases of arterial dissection in the postpartum period have been described in the literature as single case reports. We report four additional cases of internal carotid and vertebral artery dissection in the puerperal period, documented by angiography. Physicians should consider the possibility of arterial dissection in any young patient presenting with acute ischemic stroke, including women in the postpartum period. The availability of modern noninvasive ultrasound and imaging techniques may result in earlier diagnosis and facilitate identification of this condition. PMID- 10460451 TI - Absence of mutations in ATM, the gene responsible for ataxia telangiectasia in patients with cerebellar ataxia. AB - Ataxia-telangiectasia (AT) is an autosomal recessive multisystem disorder presenting in childhood with progressive cerebellar ataxia, oculocutaneous telangiectasia, immune deficiency, radiosensitivity, and cancer predisposition. The gene for AT, designated ATM (AT, mutated) encodes a protein with a carboxy terminal phosphoinositide-3 kinase domain which is involved in cell cycle checkpoints and other responses to genotoxic stress. Most of the patients with the classical AT phenotype are homozygous or compound heterozygous for severe mutations causing truncation or destabilization of the ATM protein. Patients with a milder forms of disease, called AT variants, have been found to be either homozygous for milder mutations or compound heterozygotes for null alleles and mild mutations. In order to define the clinical phenotype of patients homozygous (or compound heterozygotes) for other, milder mutations, we decided to search for ATM mutations in patients with either sporadic or familial idiopathic ataxia. Thirty-four patients with idiopathic cerebellar ataxia, aged 3-77 years, were screened for mutations in the ATM coding region. There were 12 familial cases. None of the patients had abnormal immunoglobulin or alpha-fetoprotein levels, and none had mutations in the ATM coding region. In this heterogeneous group of patients with cerebellar ataxia we found no mutations in the ATM gene. We conclude that mutations in the ATM gene are probably not a common cause for cerebellar ataxia other than AT. PMID- 10460452 TI - Eye opening in brain death. PMID- 10460453 TI - Remission of AIDS-associated progressive multifocal leukoencephalopathy after cidofovir therapy. PMID- 10460454 TI - A sporadic case of late-onset familial amyloid polyneuropathy with Bence-Jones proteinuria. PMID- 10460455 TI - Serial gadolinium-enhanced and magnetization transfer imaging to investigate the relationship between the duration of blood-brain barrier disruption and extent of demyelination in new multiple sclerosis lesions. PMID- 10460456 TI - Plasmapheresis and immunosuppression in stiff-man syndrome with type 1 diabetes: a 2-year study. PMID- 10460457 TI - Early cerebellar involvement on diffusion-weighted magnetic resonance images in herpes simplex encephalitis. PMID- 10460458 TI - Primary EBV-associated cauda equina lymphoma. PMID- 10460459 TI - Meningomyelitis in a case of toxocariasis with markedly isolated CSF eosinophilia and an MRI-documented thoracic cord lesion. PMID- 10460460 TI - Journal club PMID- 10460462 TI - ENS news PMID- 10460461 TI - Neuroscience news PMID- 10460463 TI - Robert Feulgen Prize Lecture 1999. Detection and amplification systems for sensitive, multiple-target DNA and RNA in situ hybridization: looking inside cells with a spectrum of colors. AB - In situ hybridization (ISH) is a powerful technique for localizing specific nucleic acid sequences (DNA, RNA) in microscopic preparations of tissues, cells, chromosomes, and linear DNA fibers. To date, a wide variety of research and diagnostic applications of ISH have been described, making the technique an integral part of studies concerning gene mapping, gene expression, RNA processing and transport, the three-dimensional organization of the nucleus, tumor genetics, microbial infections, and prenatal diagnosis. In this review, I first describe the ISH procedure in short and then focus on the currently available non radioactive probe-labeling and cytochemical detection methodologies that are utilized to visualize one or multiple different nucleic acid targets in situ with different colors. Special emphasis is placed on the procedures applying fluorescence and brightfield microscopy, the simultaneous detection of nucleic acids and proteins by combined ISH and immunocytochemistry, and, in addition, on the recent progress that has been made with the introduction of signal amplification procedures to increase the detection sensitivity of ISH. Finally, a comparison of fluorescence, enzyme cytochemical, and colloidal gold silver probe detection systems will be presented, and possible future directions of in situ nucleic acid detection will be discussed. PMID- 10460464 TI - Hypertrophy of growth plate chondrocytes in vivo is accompanied by modulations in the activity state and surface area of their cytoplasmic organelles. AB - The rate of longitudinal bone growth is regulated primarily by modulations in the activity of epiphyseal plate hypertrophic chondrocytes, these being manifested as changes in cell and matrix volume. It was the purpose of this study to ascertain whether the cytoplasmic organelles representing the cellular production apparatus, i.e. rough endoplasmic reticulum, Golgi apparatus and mitochondria, contribute to these changes by modulating their rate of activity or by increasing/decreasing the surface area and/or volume of their membranes. Using rats at different stages of growth, the surface areas and volumes of the three organellar systems were quantified in epiphyseal plate chondrocytes at the onset and termination of hypertrophy by ultrastructural stereology. Matrix synthesis during the same span was assessed by monitoring the production of its principal components, namely, fibrillar collagen (ultrastructural morphometry) and glycosaminoglycans (quantitative (35)S-autoradiography). Each organelle adapts to increases (21- to 35-day-old rats) and decreases (35- to 80-day-old rats) in growth rate by its own individual combination of the two alternative mechanisms, but modulations in the level of activity predominate over alterations in the surface area or volume of their membranes. These findings point to the danger of relying solely on data gleaned from a quantitative ultrastructural analysis of organellar parameters and emphasise the necessity of conducting functional assays in parallel, as performed here. PMID- 10460465 TI - Decalcification by ascorbic acid for immuno- and affinohistochemical techniques on the inner ear. AB - An ascorbic acid decalcifying solution was applied to immuno- and affinohistochemical studies on the inner ear. Rat inner ears fixed in 4% paraformaldehyde in PBS or in 2% acetic acid in ethanol solutions were adequately decalcified in an ascorbic acid solution, at a temperature of 4 degrees C. The decalcifying solution was prepared with 1% ascorbic acid and 0.84% sodium chloride in distilled water (pH 2.5-2.6). The decalcification time was in a direct relationship to the specimen calcification. In this study, two neuroactive substances (gamma-aminobutyric acid and calcitonin gene-related peptide), neurofilaments, and the galectine endogenous lectin were successfully detected immunohistochemically. PMID- 10460466 TI - Expression of platelet-derived growth factor proteins and their receptor alpha and beta mRNAs during fracture healing in the normal mouse. AB - Platelet-derived growth factor (PDGF), abundant in bone tissue, has been reported to stimulate mesenchymal cell proliferation and migration. To elucidate the functional roles of PDGF during fracture healing, we investigated the expression of PDGF-A and -B chain proteins and receptor alpha and beta mRNAs in fractured mouse tibiae. Twelve-week-old male BALB/c mice were operated on to make a closed fracture on the proximal tibia. On days 2, 4, 7, 10, 14, 21, and 28 after the operation, the fractured tibiae were excised, fixed with 4% paraformaldehyde, decalcified with 20% EDTA, and embedded in paraffin to prepare 7-microm sections. Immunohistochemistry using polyclonal antibodies against human PDGF-A and -B chains was carried out by the avidin-biotin-peroxidase method. For in situ hybridization, we used digoxigenin-labeled single-stranded DNA probes specific for mouse PDGF receptors alpha and beta generated by unidirectional polymerase chain reaction. In the inflammatory phase on days 2-4 after the fracture, mesenchymal cells gathering at the fracture site expressed the PDGF-B chain and beta receptor mRNA. At the stage of cartilaginous callus formation on day 7, the immunoreactivity for PDGF-A and -B chains on proliferating and hypertrophic chondrocytes and the signals of alpha and beta receptor mRNAs on proliferating chondrocytes became manifest. At the stage of bony callus and bone remodeling on days 14-21, the predominant expression of the PDGF-B chain and beta receptor was observed on both osteoclasts and osteoblasts. On day 28, signals for PDGF ligand proteins and receptor mRNAs diminished. The coincidental localization of PDGF ligands and their receptors implies a paracrine and autocrine mechanism. Our data suggested that PDGF contributed in part to the promotion of the chondrogenic and osteogenic changes of mesenchymal cells from the early to the midphase of fracture healing; the functions mediated by the beta receptor, including cell migration, might be prerequisites to the recruitment of mesenchymal cells in the initial step and to the interaction between osteoclasts and osteoblasts in the bone remodeling phase. PMID- 10460467 TI - Distribution of adrenomedullin (AM), proadrenomedullin N-terminal 20 peptide, and AM mRNA in the rat gastric mucosa by immunocytochemistry and in situ hybridization. AB - Adrenomedullin (AM) is a novel vasorelaxant peptide isolated from pheochromocytoma. Proadrenomedullin N-terminal 20 peptide (PAMP) is a hypotensive peptide generated by posttranslational enzymatic processing of a 185-amino acid pro-AM molecule, the same precursor as AM. In this study, we investigated localizations of these peptides by immunocytochemistry and AM mRNA by non radioisotopic in situ hybridization followed by the streptavidin and biotin complex (ABC) method and catalyzed signal amplification (CSA) in the rat adrenal medulla and gastric mucosa. In the gastric mucosa, both AM- and PAMP-like immunoreactivities were found in the neuroendocrine cells, but PAMP-positive cells were more abundant than AM-positive ones. By immunoelectron microscopy, AM and PAMP were localized exclusively in the secretory granules. The distribution pattern of AM mRNA-positive cells, only a limited portion of which had AM and/or PAMP, was also similar to that of the two peptides. But AM mRNA was detected also in a few epithelial cells as well as neuroendocrine cells. The two peptides might play an important role in the control of local circulation in the rat stomach. PMID- 10460468 TI - Polyhormonal aspect of the endocrine cells of the human fetal pancreas. AB - Histological studies were performed on 30 pancreases obtained from normal human fetuses aged between the 9th and 38th week. For immunocytochemistry, the avidin biotin-peroxidase method was used to identify and colocalise insulin, glucagon, somatostatin, pancreatic polypeptide and proliferating cell nuclear antigen. In the 9th week, cells containing all investigated peptides were present. During the fetal period, two populations of endocrine cells have been distinguished, Langerhans islets and freely dispersed cells. The free cells were polyhormonal, containing insulin, glucagon, somatostatin and pancreatic polypeptide, and were localised in the walls of pancreatic ducts throughout the whole gland. During the development of the islets we have observed four stages: (1) the scattered polyhormonal cell stage (9th-10th week), (2) the immature polyhormonal islet stage (11th-15th week), (3) the insulin monohormonal core islet stage (16th-29th week), in which zonular and mantle islets are observed, and (4) the polymorphic islet stage (from the 30th week onwards), which is characterised by the presence of monohormonal cells expressing glucagon or somatostatin. Bigeminal and polar islets also appeared during this last stage. The islets consisted of an insulin core surrounded by a thick (in the part developing from the dorsal primordium) or thin rim (part of the pancreas concerned with the ventral primordium) of intermingled mono- or dihormonal glucagon-positive or somatostatin-positive cells. The most externally located polyhormonal cells exhibited a reaction for glucagon, somatostatin and pancreatic polypeptide. Apart from the above-mentioned types of islets, all arrangements observed in earlier stages were present. Proliferating cell nuclear antigen-positive cells (single in the large islets and more numerous in the smaller ones) were predominantly observed in the outermost layer. Taken together our data indicate that, during the human prenatal development of the islet, endocrine cells are able to synthesise several different hormones. Maturation of these cells involved or depended on a change from a polyhormonal to a monohormonal state and is concerned with decreasing proliferative capacity. This supports the concept of a common precursor stem cell for the hormone-producing cells of the fetal human pancreas. PMID- 10460469 TI - Early detection of staurosporine-induced apoptosis by comet and annexin V assays. AB - Comet, TUNEL, and annexin V assays were used to identify DNA fragmentation and plasma membrane alterations occurring during staurosporine-induced apoptosis in Chinese hamster ovary cells. TUNEL assay detected apoptotic cells after 6 h treatment. The occurrence of annexin V immunofluorescence staining after 1 h treatment confirms that exposure of phosphatidylserine (PS) residues is an early biochemical feature of apoptosis. According to intensity, three annexin staining patterns were distinguished, related to different steps in the apoptotic process. The detection of highly damaged cells by the comet assay after 3 h treatment occurred earlier than the detection of DNA modifications by the TUNEL assay, but later than the exposure of PS residues. However, late apoptotic cells, otherwise characterized by plasma membrane disruption and high annexin V staining, were not detected by the comet assay. In this case, comet assay modified by omitting electrophoresis (halo assay) was more sensitive for an accurate quantification of the apoptotic fraction. PMID- 10460470 TI - Vitamin D receptor ontogenesis in rat liver. AB - Vitamin D through its receptor (VDR) plays a major role in bone mineral metabolism. However, VDR is also present in a variety of cell lines as well as in numerous tissues, suggesting other functions of the hormone beyond bone metabolism and mineral homeostasis. At the liver level, it has been shown that vitamin D induces numerous changes (i.e. enzyme activity level, stimulation of some metabolic pathways and stimulation of the normal liver recovery after partial hepatectomy). However, some works did not find VDR in the liver, and also used liver tissue as a negative control of VDR gene expression. In this paper, we examined fetal, neonatal and adult rat tissues for the presence of VDR using a sensitive RT-PCR technique and immunohistochemistry. We found VDR mRNA and VDR protein in rat liver at all different periods of rat life. Thus, we suggest that some of the actions of vitamin D on liver could be mediated at the genomic level through the VDR, and that the use of this tissue as a negative control of VDR gene expression is clearly inappropriate. PMID- 10460471 TI - Expression of oligo/polyalpha2,8-linked deaminoneuraminic acid and megalin during kidney development and maturation: mutually exclusive distribution with polyalpha2,8-linked N-acetylneuraminic acid of N-CAM. AB - The expression of homopolymers of alpha2,8-linked deaminoneuraminic acid (oligo/polyalpha2,8-KDN) and of megalin during rat kidney development was investigated using immunocytochemistry and immunoblotting, and compared to homopolymers of alpha2,8-linked N-acetylneuraminic acid (polyalpha2,8-Neu5Ac) of the neural cell adhesion molecule (N-CAM). Both, oligo/polyalpha2,8-KDN and megalin were found in early proximal tubules of embryonic day 18 kidneys. In addition, megalin, but not oligo/polyalpha2,8-KDN, was detectable in late S shaped bodies and early capillary loop stages. Until postnatal day 7, oligo/polyalpha2,8-KDN and megalin immunoreactivity was present, not only in convoluted but also in straight proximal tubules, and then restricted to the convoluted part as in adult kidney. Immunoblotting revealed increasing megalin expression until postnatal week 3 of kidney development, when the level corresponded to adult kidney. Combined immunoprecipitation/immunoblot analyses showed a steady level of oligo/polyalpha2,8-KDN on megalin throughout development. This was in striking contrast to the expression of polyalpha2,8 Neu5Ac and N-CAM, which was highest in early embryonic kidney, undetectable in kidneys of 3-week-old rats, and mutually exclusive with oligo/polyalpha2,8-KDN in its distribution. These findings demonstrated the coincidence of oligo/polyalpha2,8-KDN and megalin expression and the first appearance of proximal tubules, and revealed the high degree of specialization of the biosynthetic machinery for protein polysialylation in kidney. PMID- 10460472 TI - Highly sensitive chelating agents for histochemical staining of rare earth metals. AB - In tissues, bromopyridylazo-diethylaminophenol has been found to be capable of staining very small amounts of rare earth metals, particularly praseodymium, terbium, dysprosium, holmium, ytterbium, and lutetium. Differentiation of a target metal from interfering metals was achieved using masking agents, polyphosphates and aminopolycarboxylic acids. PMID- 10460473 TI - Comparative study of shape, course, and disintegration of the rostral notochord in some vertebrates, especially humans. AB - The rostral part of the notochord reveals many peculiarities compared with the trunk mesoderm. Furthermore, its role in head formation and inductive processes in the head is not as well understood as the interaction of the trunk notochord with the spinal cord and somites. To interpret experimental and molecular biological examinations in the developing head region, exact knowledge about morphological features of the rostral notochord is fundamental. Here we show that the rostral notochord reveals variations that depend on species and individual. We describe morphological characteristics of the rostral (head) notochord in human embryos (Carnegie stages X-XIV), which are shown in semithin sections and three-dimensional graphic reconstructions. Special attention is paid to the relationship of the notochord with the prechordal mesoderm and the adenohypophysis. We propose that in the human the rostral notochordal tip terminates at Rathke's pouch, whereas in the chick prechordal mesoderm is found in between the notochordal tip and the anlage of the adenohypophysis. The behaviour of the notochord at the end of the embryonic period proper and early fetal time is shown in sagittal histological sections of 16 to 49 mm CRL human embryos. Position and disintegration of the rostral notochord is also described in embryos of cat (8-25 mm), mouse (stage 21-24 according to Theiler) and chicken (stage 22-26 HH). A synopsis reveals the different course of the notochord within, at the inner or outer side of the basioccipital cartilage. The course of rostral notochord is determined by its attachment points at the hypophysis, the pharynx or the footplate of the brain. In all species, it has an undulating course. Its rostral tip is highly coiled, and fragments or splinters are found within the anlage of the dorsum sellae. Thus, we have reasons to believe that the adenohypophysis is a hindrance for the rostral elongation of the notochord. Variable adhesions between notochord and pharyngeal epithelium are considered to be responsible for invaginations of the pharyngeal wall forming bursae pharyngeae. In contrast to other authors, we observed in the mouse that rostrally the notochord bends ventral and penetrates the chondrocranium at the level of the later synchondrosis basisphenoidale to build a bursa pharyngea. Finally, partial duplications of two human notochords are described. PMID- 10460474 TI - Involvement of long- and short-range signalling during early tendon development. AB - Tendons connect muscle to skeletal elements. Although tendons have been shown to originate from the lateral plate mesoderm, very little is known at the molecular level about how they are formed. We have found that two genes, Follistatin and Eph-A4, are expressed in regions associated with tendon formation in developing chick limbs. Follistatin is expressed near the tip of the digits and subsequently around the tendon, whereas Eph A4 transcripts were localized in a slightly more proximal region and later in the body of the tendon. Previous work has demonstrated that application of TGFbeta1 or TGFbeta2 to inter-digital regions or the removal of ectoderm in the foot plate induces ectopic cartilage formation, while removal of ectoderm or application of FGF to tips of developing digits leads to truncation. Here we show that TGFbeta1 or removal of ectoderm is also able to induce the expression of both Eph-A4 and Follistatin and that manipulations that cause truncations affect these genes. Thus cartilage and tendon development appear to be coordinated. Ectopic application of recombinant human Follistatin, an antgaonist of certain TGFbeta super-family proteins including Activin and Bmp-4, results in the loss of tendon, implicating signalling by TGFbeta super-family in the development of tendon during chick embryogenesis. Signalling by TGFbeta family members, antagonised by Noggin is known to regulate skeletal development. Thus we suggest that parallel pathways govern both skeletal and tendon patterning. PMID- 10460475 TI - Neurotransmitter characteristics of neurons projecting to the supramammillary nucleus of the rat. AB - Retrograde labelling was combined with immunohistochemistry to localize neurons containing choline acetyltransferase, gamma-aminobutyric acid (GABA), glutamate, serotonin, somatostatin, Leu-enkephalin, neurotensin, and substance P immunoreactivity in neurons projecting to the supramammillary nucleus in the rat. Injections of wheat germ agglutinin conjugated to horseradish peroxidase (WGA HRP) into the supramammillary nucleus resulted in retrogradely labelled neurons in the medial septal nucleus, the nuclei of the diagonal band of Broca, the infralimbic cortex, the medial and lateral preoptic nucleus, the subiculum, the laterodorsal tegmental nucleus, the compact subnucleus of the central superior nucleus and the dorsal raphe nucleus. In the medial septal nucleus and in the nuclei of the diagonal band of Broca, 80-85% of WGA-HRP- labelled neurons (30-40 per section) were also immunoreactive for choline acetyltransferase and small numbers of WGA-HRP-labelled neurons were immunoreactive for GABA, glutamate, neurotensin or substance P. In the medial preoptic nucleus, 85-90% of retrogradely labelled neurons (25-30 per section) were immunoreactive for somatostatin and a few WGA-HRP-labelled neurons displayed neurotensin immunoreactivity. In the rostroventral part of the subiculum, small numbers of retrogradely labelled neurons were also immunoreactive for neurotensin or for glutamate. In the laterodorsal tegmental nucleus, 90% of WGA-HRP-labelled neurons (20-25 per section) were immunoreactive for choline acetyltransferase and small numbers of retrogradely labelled neurons also displayed substance P immunoreactivity. In the compact subnucleus of the central superior nucleus, 50 60% of retrogradely labelled neurons (15-20 per section) were also immunolabelled for GABA and approximately 30-40% of WGA-HRP-labelled neurons (10-12 per section) were immunoreactive for Leu-enkephalin. The compact subnucleus of the central superior nucleus also contained small numbers of retrogradely labelled neurons that displayed neurotensin immunoreactivity. In the dorsal raphe nucleus, 80-85% of WGA-HRP- labelled neurons (30-40 per section) were also immunoreactive for serotonin and small numbers of retrogradely labelled neurons displayed neurotensin or glutamate immunoreactivity. These results suggest that the multiple neurochemicals contained in ascending and descending projections to the SuM participate in complex interactions in the transmission process of SuM neurons. PMID- 10460476 TI - The earliest invasion of macrophages into the developing brain and eye of the tree shrew Tupaia belangeri. AB - The earliest occurrence of macrophages was investigated in the brain and optic anlagen of the tree shrew Tupaia belangeri. Nineteen serially sectioned embryos, belonging to five phases of programmed neuroepithelial cell death previously found during optic cup formation, were used. Macrophages were identified by structural criteria and by labelling with the lectin Griffonia simplicifolia I B(4). Macrophages, most probably derived from the yolk sac, are present in the perineural vessels of the phase 1 embryo (V-shaped optic evagination). Within this compartment, their number increases up to phase 4 (advanced invagination) and drops during phase 5. This first wave of macrophages is followed by a second one occurring within the perineural mesenchyme and within the neuroepithelium of the brain and eyes from phase 3 onwards. In the phase 4 embryos, a considerable rise in the number of intraventricular macrophages is noted. During phase 5 (far advanced invagination), marked vascularization of the brain starts, and a peak of macrophages is noted in the neuroepithelium and in the ventricular lumen of the brain. This spatiotemporal pattern suggests that, in Tupaia, the earliest macrophages are simultaneously shifted from perineural vessels into the neuroepithelial walls of the developing brain and, at earlier stages than previously described in other vertebrate species, of the eye anlagen. PMID- 10460477 TI - Regression of the hyaloid vessels and pupillary membrane of the mouse. AB - Regression of the pupillary membrane (PM) and hyaloid vessels - hyaloid arteries (HAs), tunica vasculosa lentis (TVL), and vasa hyaloidea propria (VHP) - in mice aged from 0 to 16 days was observed using stereomicroscopy and transmission electron microscopy. Whole-mount stereomicroscopy revealed that the pattern of normal developed vessels was basically the same as that reported in rats and rabbits and that the VHP and PM disappeared between 12 and 16 days and 10 and 12 days, respectively, while certain examples of the TVL and HA remained even at 16 days. In the TVL, VHP and PM, regression occurred segmentally and resulted in a decreased number of interconnections. The ultrastructure of the vessels in the VHP, TVL and PM was consistent with a typical capillary with pericyte covering and no fenestrations. HAs had tunica media and adventitia in the older stages. Some endothelial cells in the TVL and PM attaching to the lens capsule were thin at the side of the lens. Many macrophages were observed in the vitreous and around vessels in the whole-mount specimens at all stages. Some macrophages remained linearly arranged even after vessels became vestigial and disappeared. In transmission electron microscopy, most of these macrophages were seen to possess vacuoles and/or processes, and some of them had phagosomes. Electron microscopic findings from regressing ocular vessels were consistent with the apoptosis of both endothelia and pericytes. Obstruction of the vessels was noted at older stages. These results add further anatomical information to previous studies and suggest that the VHP and TVL as well as PM regress via apoptosis. The precise mechanisms of regression of hyaloid vessels and the role of macrophages remain for further studies. PMID- 10460478 TI - Axonal growth of newly formed vomeronasal receptor neurons after nerve transection. AB - Chemosensory neurons in the vomeronasal epithelium (vomeronasal neurons) regenerate following experimentally induced degeneration. Transection of the vomeronasal nerves leads to retrograde degeneration of vomeronasal neurons followed by replacement of the cell population. The projection of the axons of regenerated vomeronasal neurons was examined by horseradish peroxidase(HRP) histochemistry and electron microscopy. HRP-wheat germ agglutinin (WGA) was placed on the surface of the vomeronasal organ of the rat. Dense distribution of HRP-labeled fibers was observed in the vomeronasal nerve and glomerular layers in the accessory olfactory bulb (AOB) of the intact rat. At one week after transection, HRP-labeled fibers were not found in the AOB, and no labeled fibers could be observed on the medial surface of the olfactory bulb where the vomeronasal nerve traversed. Three weeks after transection, labeled fiber bundles were observed on the medial surface of the olfactory bulb in all animals. No labeled fibers were detected in the AOB. From 12 to 32 weeks after transection, projection of HRP-labeled fibers was identified in the AOB in 8 out of 26 rats (the incidence of projection was 30%). But the number of projection fibers on the operated side was much smaller than on the control side. Electron microscopy confirmed that the HRP-labeled terminals make synaptic contacts with neurons in the AOB. PMID- 10460479 TI - Functional analysis of articular cartilage deformation, recovery, and fluid flow following dynamic exercise in vivo. AB - The function of articular cartilage depends on the interaction between the tissue matrix and the interstitial fluid bound to the proteoglycan molecules. Mechanical loading has been shown to be involved in both the metabolic regulation of chondrocytes and in matrix degeneration. The purpose of the present study was therefore to analyze the deformation, recovery, and fluid flow in human articular cartilage after dynamic loading in vivo. The patellae of 7 volunteers were imaged at physical rest and after performing knee bends, with a specifically optimized fat-suppressed FLASH-3D magnetic resonance (MR) sequence. To measure cartilage deformation, the total volume of the patellar cartilage was determined, employing 3D digital image analysis. Patellar cartilage deformation ranged from 2.4 to 8.6% after 50 knee bends, and from 2.4% to 8.5% after 100 knee bends. Repeated sets of dynamic exercise at intervals of 15 min did not cause further deformation. After 100 knee bends, the cartilage required more than 90 min to recover from loading. The rate of fluid flow during relaxation ranged from 1.1 to 3.5 mm(3)/min (0.08 to 0.22 mm(3)/min per square centimeter of the articular surface) and was highly correlated with the individual degree of deformation after knee bends. The data provide the first quantification of articular cartilage recovery and of the rate of fluid flow between the cartilage matrix and surrounding tissue in intact joints in vivo. Measurement in the living opens the possibility of relating interindividual variations of mechanical cartilage properties to the susceptibility of developing joint failure, to assess the load-partitioning between the fluid phase and solid cartilage matrix during load transfer, and to determine the role of mechanically induced fluid flow in the regulation of the metabolic activity of chondrocytes. PMID- 10460480 TI - Pulmonary lymphatic filling is increased in spontaneously hypertensive rats. AB - Extravascular lung liquid must rely on tissue-space pressure gradients to drive it into the lymphatics because the fluid is outside the lymphatic contractile pumping and valve control. Focal tissue pressure changes could result from muscular contraction in the blood vessel walls. Perivascular lymphatics usually lie within the adventitia of pulmonary blood vessels, and are generally more noticeable in veins than arteries. Spontaneously hypertensive rats have exaggerated focal pulmonary venous muscle (venous sphincters). These muscular tufts are often near initial lymphatics; if their contraction was important for lymph transport, spontaneously hypertensive rats could have more lymphatic filling in the areas of the pulmonary venous sphincters than normotensive rats. Because the focal muscularity is found in pulmonary veins more than arteries, veins may have more focal lymphatic filling than arteries. To test these hypotheses, lung histology and vascular and lymphatic casts of spontaneously hypertensive and normotensive rats were examined. Contracted venous sphincters were found on 108 of 127 veins with lymphatics in the spontaneously hypertensive rats and 5 of 41 in the normotensive rats P<0.01). The spontaneously hypertensive rats had deeper venous contractions and more lymphatic filling around both arteries and veins (P<0.01). In the hypertensive rats, the venous was greater than the arterial lymphatic filling (P<0.01). On the pleural surface, hypertensive rats also had greater lymphatic filling than controls (P<0.01). This anatomic evidence suggests that pulmonary venous sphincters are associated with focal lymphatic filling, and perivascular muscle action might be a component of the pulmonary lymphatic system. PMID- 10460482 TI - Cell and tissue research announces online first publication PMID- 10460481 TI - Development of the thymus, spleen, lymph nodes and liver in the marsupial, Isoodon macrourus (Northern brown bandicoot, Peramelidae). AB - We report for the first time the development and morphological characteristics of the spleen thymus, lymph nodes and liver of the northern brown bandicoot, Isoodon macrourus. To date few marsupial species have been studied. The development and morphological characteristics of the organs shared the typical features of those few other marsupials studied as well as those of eutherians. These suggest comparative functional properties with the eutherian immune system. The thymus differentiated within the first week of pouch life and showed evidence of immunolymphopoiesis up to juvenile stages. The spleen, though seeded by lymphocytes within the first week of pouch life, was slower to mature, but differentiated and showed signs of immunocompetency by the time young left the pouch. The mature spleen displayed the same anatomical blood filtering and immunosurveillance properties as that of the eutherian spleen, with evidence of erythrocyte destruction, thrombopoietic activity, activation and differentiation of immunocompetent lymphocytes. However, the absence of sheathed capillary structures in the spleen may indicate differences in the humoral response to circulating antigens. Similarly, lymph nodes also mature by this stage with anterior nodes appearing before posterior nodes. The mature lymph nodes displayed structural features of secondary immuno-lymphoid organs consistent with production of immune responses. Finally, the liver displayed haemopoietic activity for the first four weeks of pouch life. The pattern of development in the bandicoot appears to parallel the pattern reported for other marsupials, yet the thymus matures considerably earlier than previously reported and may be of significance in the development and onset of cell-mediated immunity. Current studies to characterise cellular components, such as T/B lymphocyes and accessory cells of these organs will help to define the mechanisms of immune recognition, activation and hence outline the basis of the marsupial humoral and cellular immunity. PMID- 10460483 TI - Development of the cholinergic neurotransmitter phenotype in postganglionic sympathetic neurons. AB - Sympathetic ganglia are composed of noradrenergic neurons and cholinergic neurons that differ in the expression of neurotransmitter-synthesizing enzymes, neurotransmitter transporters and neuropeptides. The analysis of the cholinergic differentiation during development revealed important principles involved in the generation of neuronal diversity, in particular the importance of signals from the innervated target. Some peripheral targets, such as the sweat glands in the mammalian footpads, are purely cholinergically innervated in the adult, whereas skeletal muscle arteries receive both noradrenergic and cholinergic innervation. For sympathetic neurons innervating sweat glands there is convincing evidence that these neurons are initially noradrenergic and that the interaction of innervating fibers and target tissue induces a shift in the neurotransmitter phenotype from noradrenergic to cholinergic. In addition to this target-dependent differentiation, an earlier expression of cholinergic characters was observed in sympathetic ganglia that occurs before target contact. These data raise the possibility that different subpopulations of cholinergic sympathetic neurons, innervating distinct peripheral targets, may develop along distinct schedules. In vitro studies suggest that growth factors of the family of neuropoietic cytokines are involved in the specification of the cholinergic sympathetic phenotype. Recent in vivo studies that interfered with cytokine receptor expression in developing avian sympathetic ganglia indicate that only the late, target dependent differentiation depends on cytokine signaling. The signals involved in the early, target-independent expression of cholinergic properties remain to be determined, as well as the identity of the target-derived cytokine. Thus, cholinergic sympathetic differentiation seems to be more complex than expected, involving either both target-independent and target-dependent control or only target-induced differentiation, according to the specific neuronal subpopulation and target. PMID- 10460484 TI - Molecular and functional aspects of latent transforming growth factor-beta binding protein: just a masking protein? AB - Latent transforming growth factor-beta (TGFbeta) binding protein (LTBP), a component of the high-molecular-weight latent TGFbeta complex, is found in various cell and tissue types. Originally described as a TGFbeta-masking protein, recent detections of four isoforms and numerous splice variants provide new aspects of its putative functional role. Regulation and sequestration of TGFbeta activity and structural remodeling of the extracellular matrix (ECM) seem to be the main tasks, but other possible functions might exist. The mechanism by which LTBP interacts with cell surface molecules or cellular receptors and ECM components remains unclear. Cellular, molecular and functional aspects will be discussed. PMID- 10460485 TI - Genesis, a Winged Helix transcriptional repressor, has embryonic expression limited to the neural crest, and stimulates proliferation in vitro in a neural development model. AB - A novel repressor of the Winged Helix (formerly HNF-3/Forkhead) transcriptional regulatory family, termed Genesis (also called HFH2), was previously found to be exclusively expressed in primitive embryonic cell lines. In this study in situ cRNA hybridization experiments revealed that Genesis was expressed during embryogenesis only in developing neural crest cells. Its expression diminished upon their terminal differentiation into sympathetic and parasympathetic neurons. Based on that finding, Genesis was retrovirally transduced into pluripotent N Tera-2 clone D1 (NT2/D1) teratocarcinoma cells, which are a well-described in vitro model of neural development. Retinoic acid (RA) treatment will drive these cells to differentiation toward the neuronal lineage and cause an increase in expression of the cyclin-dependent kinase inhibitor p21 protein, which leads to an inhibition in cellular proliferation. Although RA-induced expression of neuronal differentiation markers was not influenced by forced overexpression of Genesis in NT2-D1 cells, proliferation of Genesis-transduced cells continued following RA treatment. RA was unable to induce the expression of the cyclin dependent kinase inhibitor p21 in the Genesis-transduced cells, but Go/G1 tumor suppressor p53 expression was induced normally. Therefore, Genesis may play a role in the regulation of primitive neural crest development by preventing terminal quiescence through inhibition of p21 protein expression. These data also lend evidence for the hypothesis that proliferation and differentiation pathways are not irrevocably linked, but can function independently. PMID- 10460486 TI - Differential antigen expression during metamorphosis in the tripartite olfactory system of the African clawed frog, Xenopus laevis. AB - In the adult African clawed frog, Xenopus laevis, olfactory epithelium is housed in three separate nasal cavities: the principal cavity, the middle cavity, and the vomeronasal organ. The sensory epithelium in each of these cavities has distinct cellular features, and presumed physiological and behavioral functions, which arise during metamorphosis. Most notably, the middle cavity is formed de novo, and the principal cavity is transformed from a larval sensory epithelium with water exposure to an adult olfactory epithelium with air exposure. To understand the cellular nature of this plasticity more clearly, we characterized the staining patterns generated in the olfactory system of X. laevis with a new monoclonal antibody, anti-E7. The olfactory epithelium is first stained with anti E7 during late embryonic development. Transection of the olfactory nerves during metamorphosis eliminates all staining and indicates that the staining is associated with mature or nearly mature olfactory receptor neurons. The antibody diffusely stains the vomeronasal organ throughout development and in adults. In the larval principal cavity, the olfactory receptor neurons are brightly stained, but this cellular staining is lost after metamorphosis. The mucus from Bowman's glands in the principal cavity, however, is intensely stained in adults. The middle cavity, throughout development and in adulthood, has the same staining characteristics as the larval principal cavity. Thus, the E7 antibody can distinguish the three areas of the olfactory epithelium, allowing measurement of sensory epithelium volume, and serves as an excellent marker for the changes in the sensory epithelium that occur during metamorphosis. PMID- 10460487 TI - Morphology and synaptic connectivity of nitric oxide synthase-immunoreactive neurons in the guinea pig retina. AB - Immunocytochemical methods with an antiserum against neuronal nitric oxide synthase (NOS) were applied to identify the morphology and synaptic connectivity of NOS-like immunoreactive neurons in the guinea pig retina. In the present study, two types of amacrine cells were labeled with anti-NOS antisera. Type 1 cells had large somata located in the inner nuclear layer (INL) with long, sparsely branched processes ramifying mainly in stratum 3 of the inner plexiform layer (IPL). The somata of type 2 cells (smaller diameters) were located in the INL. Some displaced amacrine cells in the ganglion cell layer were labeled. The soma size of the displaced amacrine cells was similar to that of the type 2 amacrine cells. However, processes originating from type 2 amacrine cells and displaced amacrine cells stratified mainly in strata 1 and 5, respectively. Some cone bipolar cells were weakly NOS-immunoreactive. The synaptic connectivity of NOS-like immunoreactive amacrine cells was identified in the IPL by electron microscopy. NOS-labeled amacrine cell processes received synaptic input from other amacrine cell processes and bipolar cell axon terminals in all strata of the IPL. The most frequent postsynaptic targets of NOS-immunoreactive amacrine cells were other amacrine cell processes. Cone bipolar cells were postsynaptic to NOS-labeled amacrine cells in all strata of the IPL. Labeled amacrine cells synapsing onto ganglion cells were found only in sublamina b. A few synaptic contacts were observed between labeled cell processes. In the outer plexiform layer, dendrites of labeled bipolar cells made basal contact with cone pedicles or formed a synaptic triad opposed to a synaptic ribbon of cone pedicles. PMID- 10460488 TI - Distribution of intrinsic cardiac neurons in whole-mount guinea pig atria identified by multiple neurochemical coding. A confocal microscope study. AB - Functional data indicate that neurons in distinct regions of the heart exert preferential regional cardiac control. To date the regional distribution of specific types of neurons within the intrinsic cardiac nervous system remains unknown, as does their associations with distinct neurotransmitter and/or neuromodulatory profiles. This study was designed to ascertain: (1) the distribution of different classes of neurons within the intrinsic cardiac nervous system as determined by microscopic analysis; (2) the neurochemical profiles of neurons in differing atrial loci; (3) which neurochemicals are co-localized within specific populations of intrinsic cardiac neurons; and (4) the distribution of specific sub-populations of neurons expressing specific immunoreactivities. Taking advantage of confocal laser scanning microscopy and distinct immunoreactive fluorescent markers in various double-label combinations, several sub-populations of intrinsic cardiac neurons were identified. Of all identified neurons, 85-90% were located in ganglia (ganglionic neurons), the rest being isolated (individual neurons). The two general neuronal markers protein gene product 9.5 (PGP 9.5) and microtubule-associated protein (MAP-2) were associated with neurons clustered primarily in the interatrial septum and around the origins of the two vena cavae. Ganglia (group 1) contained three sub populations of neurons: approx. 80% of ganglionic neurons were large (15-40 microm diameters; group 1a) and approx. 20% had smaller diameters (less than 15 microm; group 1b). All of these neurons were PGP-immunoreactive, exhibiting choline acetyltransferase (ChAT) immunoreactivity (IR), tyrosine hydroxylase (TH) IR, neuropeptide Y (NPY) IR, vasoactive peptide (VIP) IR and substance P (SP) IR. The remaining 5% of ganglionic neurons were small (group 1c; less than 20 microm). These displayed TH immunoreactivity but not MAP, PGP, CHAT, NPY or SP immunoreactivity. Ten to fifteen percent of all neurons loosely distributed outside of ganglia were small (10-25 microm) and located primarily around the origin of the superior vena cava. They displayed immunoreactivity to TH, ChAT, VIP, NPY and SP, but not to MAP-2 or PGP 9.5. These data provide anatomical and immunohistochemical evidence for specific localization of differing populations of intrinsic cardiac neurons with respect to their size, ganglionic distributions and capacity to express multiple neurotransmitters. Although the functional importance of such a regional distribution of differing populations of intrinsic cardiac neurons remains unknown, these anatomical data support the thesis that unique clustering of specific populations of neurons within this nervous system represents the anatomical substrate for complex local cardiac regulatory phenomena occurring at the level of the target organ. PMID- 10460489 TI - Relationships between nerves and myofibroblasts during cutaneous wound healing in the developing rat. AB - Contraction of skin excision wounds is affected by age and the presence of peripheral nerves. The present study examined relationships between peripheral innervation, wound contractile cells, and rate of wound closure to determine whether these are altered during development. Full-thickness 4-mm-diameter circular flaps were excised from the interscapular skin of rats on postnatal day (PND) 5, PND 12, or PND 60. Wounds of PND 5 and PND 12 rats contracted 45% between post-wound days (WD) 3 and 5 and more slowly thereafter, with a scar 9 14% of the original wound size by WD 21. In contrast, PND 60 wounds contracted only 22% between WD 3 and 5, and the residual scar at WD 21 was 40% of the original wound size. In younger rats, alpha-smooth muscle actin-immunoreactive myofibroblasts first appeared on WD 2 and attained maximum density at WD 5. Innervation, as assessed by protein gene product 9.5 immunoreactivity, appeared by WD 3 and increased rapidly through WD 7 in younger rats. In PND 60 wounds, myofibroblasts did not appear until WD 5 and did not attain a maximum until day 10. Nerve ingrowth was not significant until WD 10 and was depressed relative to younger rats throughout the healing phase. Wound nerves were predominantly immunoreactive to calcitonin gene-related peptide, and synaptophysin immunostaining revealed close associations between varicosities and myofibroblasts. These findings suggest that wound myofibroblasts may be a target of peripheral nerves, and delayed wound closure in mature rats is associated with deficiencies in both myofibroblasts and innervation. PMID- 10460490 TI - Early stages of endothelial wound repair: conversion of quiescent to migrating endothelial cells involves tyrosine phosphorylation and actin microfilament reorganization. AB - Endothelial repair to reestablish structural integrity following wounding is a complex process. Since the actin cytoskeleton undergoes specific changes in distribution as quiescent endothelial cells switch to activated migrating cells over a 6-h period following wounding (Lee et al. 1996), we studied tyrosine phosphorylation in association with actin microfilaments and adhesion proteins using double immunofluorescent confocal microscopy. We showed that in a confluent monolayer phosphotyrosine localized at the periphery of the cell at vinculin cell cell adhesion sites within the actin-dense peripheral band (DPB) and centrally at talin/vinculin cell-substratum adhesion sites at the ends of central microfilaments. Over a period of 6 h following in vitro wounding there was a reduction of peripheral phosphotyrosine associated with the loss of both cell cell adhesion sites and the DPB (stage I). Concomitantly, an increase in central phosphotyrosine was associated with an increase in cell-substratum adhesion sites and central microfilaments parallel to the wound edge (stage II), which subsequently redistributed perpendicular to the wound edge (stage III). We also localized FAK and paxillin at the ends of parallel and perpendicular central microfilaments. Immunoprecipitation of paxillin showed increased phosphotyrosine and protein levels when prominent central microfilaments were present and underwent remodeling. Inhibition of tyrosine kinases by genistein and tyrosine phosphatases by sodium orthovanadate resulted in reduced endothelial repair associated with disruption of adhesion site formation and central microfilament formation/redistribution in each stage of repair. We suggest that tyrosine phosphorylation of adhesion proteins, such as paxillin, may be important in regulating the early stages of endothelial wound repair. PMID- 10460491 TI - Correlation between gender and spontaneous C-cell tumors in the thyroid gland of the Wistar rat. AB - In many rat strains, C-cell hyperplasia occurs in an age-dependent manner and is often associated with multifocal C-cell carcinoma. The purpose of this study was to investigate the spectrum of spontaneous, proliferative C-cell disorders by gender in Wistar rats throughout their lifespan. The incidence of C-cell hyperplasia shows a significant increase with age (P<0.001) and is much higher in female rats than in male rats (P<0.05). From 3 to 24 months of life, 27.5% of female rats showed a normal C-cell pattern, 55.0% showed C-cell hyperplasia, and 17.5% showed C-cell tumors; while 57.5% of male rats showed a normal C-cell pattern, 32.5% showed C-cell hyperplasia, and 10% showed C-cell tumors. Although the overall frequency of C-cell neoplasms in females was nearly double that in males, these data are not statistically significant. However, the number of C cell tumors showed a significant increase with age (P<0.05). Therefore, we can conclude that there were significant differences in the incidence of the total spectrum of C-cell proliferative abnormalities in the thyroid gland of Wistar rats that were both age-dependent and gender-dependent. PMID- 10460492 TI - Evidence for gonadotropin-releasing hormone receptor mRNA expression by estrogen in rat granulosa cells. AB - The hormonal regulation of ovarian gonadotropin-releasing hormone (GnRH) receptor mRNA expression has been examined by in situ hybridization in hypophysectomized immature rats. In hypophysectomized rats, GnRH receptor mRNA expression is localized in the interstitial cells. After diethylstilbestrol treatment, most follicles grow to form early antral follicles and express GnRH receptor mRNA in the peripheral part of the granulosa layer, indicating that the expression in the growing follicles is estrogen-dependent. Only weak or no expression of the receptor mRNA is detectable in the atretic follicles of hypophysectomized rats, whereas very strong expression has been observed in the granulosa cells of atretic follicles of intact immature rats. Administration of testosterone or a GnRH agonist, both of which are atretic agents for ovarian follicles, to hypophysectomized rats markedly increases the apoptotic cell death of the granulosa cells but fails to induce GnRH receptor mRNA expression. The co administration of these agents with diethylstilbestrol causes the granulosa cells of atretic follicles to express the receptor mRNA very strongly, suggesting that this mRNA expression in the atretic follicles is also estrogen-dependent. On the other hand, expression of the receptor mRNA in the ovarian interstitial cells is not affected by hypophysectomy or hormone treatments. All of these results clearly indicate that estrogen is essential for the expression of ovarian GnRH receptor mRNA in the granulosa cells of atretic follicles and growing follicles, whereas the expression in the interstitial cells is estrogen-independent. PMID- 10460493 TI - Intracellular production of interleukin-18 in human epithelial-like cell lines is enhanced by hyperosmotic stress in vitro. AB - Interleukin-18 is a novel multifunctional cytokine, which enhances natural killer cell activity and promotes the induction of cytokine production, including that of interferon-gamma by T cells and antitumor effects. Interleukin-18 is produced by cells of several different tissues (e.g., macrophages, keratinocytes, osteoblasts, and intestinal epithelium); however, it is unclear what physiological conditions or stimuli induce interleukin-18 production. To determine physiological conditions for the production of interleukin-18, we have examined the effect of mannitol-induced hyperosmotic conditions on normal human umbilical vein endothelial cells (HUVEC) and eight established human epithelial like cell lines (Intestine 407, Caco-2, A253, HeLa, SCC25, HT1197, ACHN, A549). Hyperosmotic conditions induced interleukin-18 immunoreactivity in all the human cell lines tested, as detected by immunocytochemistry. The enhanced interleukin 18 production was also observed when mannitol was replaced with NaCl as the inducer of hyperosmotic stress. Enzyme-linked immunosorbent assays revealed that interleukin-18 concentrations in cell extracts were significantly increased by hyperosmotic conditions. Reporter gene assays also revealed that hyperosmotic conditions stimulated transcriptional activity of the interleukin-18 promoter. These results show for the first time that hyperosmotic stress is a stimulator of interleukin-18 production in epithelial-like cells. PMID- 10460494 TI - Differential in vitro phenotype pattern, transforming growth factor-beta(1) activity and mRNA expression of transforming growth factor-beta(1) in Apert osteoblasts. AB - The phenotype of Apert osteoblasts differs from that of normal osteoblasts in the accumulation of macromolecules in the extracellular matrix. Apert osteoblasts increase type I collagen, fibronectin and glycosaminoglycans secretion compared with normal osteoblasts. Because the extracellular matrix macromolecule accumulation is greatly modulated by transforming growth factor-beta(1), we examined the ability of normal and Apert osteoblasts to secrete transforming growth factor-beta(1) by CCL-64 assay and to produce transforming growth factor beta(1 )by analysis of the mRNA expression of transforming growth factor-beta(1). Northern blot analysis revealed an increased amount of transforming growth factor beta(1) mRNA expression in Apert osteoblasts compared with normal ones. Moreover, the level of the active transforming growth factor-beta(1) isoform was higher in Apert than in normal media. In pathologic cells, the increase in transforming growth factor-beta(1) gene expression was associated with a parallel increase in the factor secreted into the medium. The level of transforming growth factor beta(1) was decreased by the addition of basic fibroblast growth factor. Transforming growth factor-beta(1) is controlled temporally and spatially during skeletal tissue development and produces complex stimulatory and inhibitory changes in osteoblast functions. We hypothesise that in vitro differences between normal and Apert osteoblasts may be correlated to different transforming growth factor-beta(1) cascade patterns, probably due to an altered balance between transforming growth factor-beta(1) and basic fibroblast growth factor. PMID- 10460495 TI - Localization and ultrastructure of pulmonary dendritic cells during delayed-type hypersensitivity reactions in mice. AB - Dendritic cells (DCs) are widely distributed in the airways and can serve as potent antigen-presenting cells. To clarify their involvement in the cell mediated immune responses of the lung, we immunohistochemically investigated their distribution and kinetics during pulmonary delayed-type hypersensitivity (DTH) reactions induced in sensitized mice by intratracheal instillation of hapten. Cellular infiltrate appeared around the bronchiole and its accompanying blood vessel at 12 h after elicitation and progressively expanded by 48 h. As quantitated by computer-assisted morphometry, I-A(+) DCs and CD4(+) Th cells significantly increased in number around the bronchiole to a maximum at 24 h, whereas F4/80(+) macrophages were predominantly accumulated around the accompanying vessel with a peak at 48 h. Serial-section analysis revealed that DCs were colocalized with Th cells in the inflamed peribronchiolar tissue. Immunoelectron microscopy demonstrated that DCs found inside and around the capillaries and venules of peribronchiolar interstitium displayed round forms, indicating their emigration from here, while those situated far from the microvessels were elongated, often in close apposition to the lymphocytes. Mitosis of DCs was rarely seen. The present results suggest that peribronchiolar accumulation of DCs resulting from accelerated influx of blood-borne immature DCs and the interaction with T cells at the application site may play inducing roles in the development of pulmonary DTH reactions by enhancing the recruitment of macrophages. PMID- 10460496 TI - Ultrastructural, immunocytochemical and morphometric characterization of liver peroxisomes in gray mullet, Mugil cephalus. AB - Peroxisomes of the hepatocytes of gray mullets, Mugil cephalus, were characterized cytochemically and immunocytochemically using antibodies against the peroxisomal proteins catalase and palmitoyl-coenzyme A (CoA) oxidase. In addition, morphometric parameters of peroxisomes were investigated depending on the hepatic zonation, the age of the animals and the sampling season. Mullet liver peroxisomes were reactive for diaminobenzidine, but presented a marked heterogeneity in staining intensity. Most of the peroxisomes were spherical or oval in shape, although irregular forms were also observed. Their size was heterogeneous, with profile diameters ranging from 0.2 to 3 microm. Peroxisomes tended to occur in clusters, usually near the mitochondria and lipid droplets. They also showed a very close topographical relationship to the smooth endoplasmic reticulum. Mullet liver peroxisomes did not contain cores or nucleoids as rodent liver peroxisomes, but internal substructures were observed in the matrix, consisting of small tubules about 60 nm in diameter and larger semicircles 120 nm in diameter. The volume density of peroxisomes was higher in periportal hepatocytes of mullets sampled in summer than in pericentral hepatocytes, indicating that mullet peroxisomes vary depending on physiological and environmental conditions. By immunoblotting, the mammalian antibodies cross react with the corresponding proteins in whole homogenates of mullet liver. Paraffin sections immunostained with the antibodies against catalase and palmitoyl-CoA oxidase showed a positive reaction corresponding to peroxisomes localized in the hepatocyte cytoplasm. In agreement, the ultrastructural study revealed that catalase and palmitoyl-CoA oxidase are exclusively localized in the peroxisomal matrix in fish hepatocytes, showing a dense gold labeling. The presence of the peroxisomal beta-oxidation enzyme palmitoyl-CoA oxidase in peroxisomes indicated that these organelles play a key role in the lipid metabolism of fish liver. PMID- 10460497 TI - Immunohistochemical localization of calbindin D28k during root formation of rat molar teeth. AB - The present study was undertaken to examine the localization of calbindin D28k (CB)-like immunoreactivity (-LI) during the root formation of the rat molar. In the adult rat, CB-LI was detected in some of the cells of the epithelial rest of Malassez at the bifurcational region and in certain cells between the root dentin and cementum at the apical region. These cells had indented nuclei and many tonofilaments, and cementocytes lacked CB-LI. Moreover, CB-LI was observed in the periodontal fibroblasts in the alveolar half of the apical region. During root formation, the cells in the Hertwig's epithelial root sheath (HERS) lacked CB-LI, but most fragmented cells along the root surface began to express CB-LI when HERS was disrupted. Preodontoblasts and odontoblasts at the apical portion of the root also showed CB-LI. After the formation of cellular cementum, the CB immunoreactive (-IR) cells were entrapped between the root dentin and cementum in the apical portion of the root. The number of CB-IR cells at the root surface decreased gradually, while that between the root dentin and cementum increased. The fibroblasts in the periodontal ligament began to express CB-LI after commencement of the occlusion, and the number and the staining intensity of CB-IR fibroblasts increased gradually with the passage of time. The present results suggest that CB may play an important role in the survival of the epithelial cells, in the cellular responses of periodontal fibroblasts against mechanical forces caused by the occlusion, and in the initial mineralization by the odontoblasts through the regulation of intracellular Ca(2+) concentration. PMID- 10460498 TI - Isolation and characterization of an apically sorted 41-kDa protein from the midgut of tobacco hornworm (Manduca sexta). AB - Immunocytochemical localization and sorting properties of a newly purified 41-kDa protein (MsM41) were investigated in an insect, the tobacco hornworm Manduca sexta. The protein purified from midgut homogenates of feeding fifth-stadium larvae was found exclusively in this tissue on Western blots. Presence of MsM41 protein was indicated in both anterior and posterior regions of the midgut during the whole fifth stadium. However, in the posterior region an additional 39-kDa protein was also detected during the feeding period of the last larval stage. Upon light-microscopic examination immunoreactivity was localized in the columnar cells, while the goblet, endocrine and regenerative cells remained unlabeled. Distribution of the label during the feeding period was different in the anterior and posterior regions. In the anterior region immunoreactivity was localized only to the brush border membrane of columnar cells, while in the posterior region some cytoplasmic structures identified as large trans-Golgi vesicles, endoplasmic reticulum and small secretory vesicles were also labeled. Large, apical extrusions remained immunonegative. In vitro translation confirmed that our protein was expressed only in the posterior region of the midgut. The primary translation product was a 39-kDa protein. Putative post-translational modifications yielded the 41-kDa form, which was then secreted apically. Its presence in the region of the anterior part microvilli was probably due to the countercurrent flux of the ectoperitrophic fluid. PMID- 10460499 TI - Glial cells in the developing and adult olfactory lobe of the moth Manduca sexta. AB - The antennal lobe of the moth contains several classes of glial cells that are likely to play functional roles in both the developing and mature lobe. In this study, confocal and electron microscopy were used to examine in detail the morphology of two classes of glial cells, those associated with olfactory receptor axons as they course to their targets in the lobe and those that form borders around the synaptic neuropil of the olfactory glomeruli. The former, the nerve-layer glia, have long processes with multiple expansions that enwrap axon fascicles; the latter, the neuropil glia, constitute two subgroups: complex glia with large cell bodies and branching, vellate arbors; and simple glia, with multiple, mostly unbranched processes with many lamellate expansions along their lengths. The processes of complex glia appear to be closely associated with axon fascicles as they enter the glomeruli, while those of the simple glia surround the glomeruli as part of a multi-lamellar glial envelope, their processes rarely invading the synaptic neuropil of the body of the glomerulus. The full morphological development of antennal-lobe glial cells requires more than two thirds of metamorphic development. During this period, cells that began as cuboidal or spindle-shaped cells that were extensively dye-coupled to one another gradually assume their adult form and, at least under nonstimulated conditions, greatly reduce their coupling. These changes are only weakly dependent on the presence of olfactory receptor axons. Glial processes are somewhat shorter and less branched in the absence of these axons, but basic structure and degree of dye-coupling are unchanged. PMID- 10460500 TI - Color Doppler energy--a new technique to study tissue perfusion in renal transplants. AB - Information on renal tissue perfusion after transplantation remains important for renal allograft monitoring. Findings obtained by conventional Doppler sonography are limited to vascular resistance (RI). The new technique color Doppler energy (CDE) is Doppler angle independent, omits flow velocity and direction, and is proportional to the returning signal strength. The aim of our study was to standardize the application of this technique and to analyze the information obtained. Forty-six CDE studies were performed with an Acuson 128XP in 28 children (mean age 12.4+/-5.3 years) between 4 days and 10 years after renal transplantation. The most-reproducible information was obtained with a 5-MHz linear probe and a constant area of 2x3 cm (log compression 40 dB, filter 3). CDE provided a high-resolution cross-sectional display of perfused cortical tissue vessels. According to the density of signals, the perfusion could be grouped into six perfusion scores (PS). The interobserver concordance was more than 85%. No correlation was found between PS and RI or blood pressure. However, there was a significant correlation between PS and glomerular filtration rate (r=-0.78, P<0. 001). These first results demonstrate a significant relationship between PS and chronic rejection. Non-rejection-related functional impairment exhibited no decrease in PS. We conclude that our proposed standardized CDE renal study is observer independent. CDE is a promising new technique that provides information on renal allograft dysfunction that is different from classical color Doppler findings. Further studies will clarify its role in renal transplant monitoring and its ability to replace more-invasive techniques. PMID- 10460501 TI - The role of early renal biopsy in cyclosporin induced thrombotic microangiopathy. AB - Thrombotic microangiopathy is an uncommon complication of cyclosporin immunosuppression following renal transplantation. We present a 15-year-old girl who developed clinical features of acute rejection, but in whom an early biopsy revealed thrombotic microangiopathy, allowing a change to FK506 immunosuppression resulting in excellent graft recovery. PMID- 10460502 TI - Renal remodelling in dietary protein modified rat polycystic kidney disease. AB - Dietary protein restriction slows progression of the Han:SPRD-cy rat model of polycystic kidney disease. We undertook studies to examine the relative changes in interstitial and tubular pathology as a result of feeding an 8% casein (LP) diet to Han:SPRD-cy rats. Archival tissue from a previous study comparing LP and 20% casein (NP) diets was examined morphometrically after immunohistochemical or histochemical staining for apoptosis, proliferation antigens, interstitial fibrosis, and macrophage infiltration. Expression of common extracellular matrix genes was measured by Northern analysis. Animals fed LP diet demonstrated reduced tubular epithelial remodelling compared with animals fed NP diet by both proliferating cell nuclear antigen-positive cells (57.5 vs. 71.6 cells/mm epithelium, P=0.007) or apoptosis (31.2 vs. 35.6 cells/mm epithelium, P=0.006). Interstitial pathology demonstrated that LP feeding was associated with proportionately greater reductions in interstitial fibrosis (0.3 vs. 1.3 ml/kg body weight, P=0.003), interstitial cellularity (361 vs. 604 cells/high-power field, P=0.0002), and interstitial macrophages (67 vs. 149 cells/high-power field, P=0. 0002). Northern analysis only revealed significantly lower levels of monocyte chemoattractant protein mRNA (P=0.04) in animals fed the LP diet. Dietary protein restriction modifies both tubule and interstitium, with significant impact upon interstitial inflammation and fibrosis in the Han:SPRD-cy rat. PMID- 10460503 TI - Longitudinal evaluation of transport kinetics in children receiving peritoneal dialysis. AB - Functional stability of the peritoneal membrane is necessary for maintenance of peritoneal dialysis (PD) as a therapeutic option. Few studies have investigated this issue in children. We evaluated the peritoneal membrane solute transport capacity longitudinally in 26 children (mean age 11.0+/-5.5 years) receiving long term PD. Each patient underwent a standardized peritoneal equilibration test on two occasions (mean interval between studies 19.8+/-5.9 months) to determine solute dialysate to plasma (D/P) ratios, dialysate glucose to initial dialysate glucose (D/D(0)) ratios, and mass transfer area coefficients (MTAC). The correlation of transport capacity with peritonitis history was also assessed. No significant change in MTAC, D/P, or D/D(0) values were found when comparing original and follow-up data of the group overall. However, transport of creatinine and glucose was significantly (P<0.05) greater in the peritonitis group compared with the group without peritonitis, and differences in the change over time between the peritonitis groups was significant for MTAC creatinine (P=0.035) and D/D(0) glucose (P=0.020). In summary, this experience demonstrates functional stability of the peritoneal membrane in pediatric patients receiving PD. However, follow-up assessments of peritoneal solute kinetics may be necessary in patients with a history of peritonitis in order to permit early identification of those who may be at risk for ultrafiltration failure and sclerosing peritonitis. PMID- 10460504 TI - Tuberculosis in children undergoing continuous ambulatory peritoneal dialysis. AB - The incidence of tuberculosis (TB) is increasing worldwide. Due to an impairment of cellular immunity, patients with chronic renal failure are susceptible to reactivation of TB. Seventy patients were treated by continuous ambulatory peritoneal dialysis (CAPD) in our pediatric nephrology department during the years 1989-1997. TB was diagnosed in 4 patients, representing 5.7% of all CAPD patients in our department. One patient had extrapulmonary (TB osteomyelitis) and the others had pulmonary TB. All patients were treated with antituberculous drugs. Two patients with pulmonary TB were cured. Symptoms improved in the other 2 patients but they died at home for unknown reasons. We recommend that all children in regions of high prevalence of TB should be investigated for TB, especially if they have a cough or fever of unknown etiology. PMID- 10460505 TI - Intravenous iron treatment of renal anemia in children on hemodialysis. AB - Treatment of anemia in children with end-stage renal disease (ESRD) has been greatly facilitated by the introduction of recombinant human erythropoietin (rHuEPO). A major limiting factor in the treatment of renal anemia is sufficient iron supplementation. Eight children (aged 10-17 years) receiving hemodialysis were treated with intravenous iron (1 mg/kg per week) for 3 months. Hemoglobin (Hb), hematocrit (Hct), and serum ferritin levels were measured regularly. The mean Hct increased from 25% to 30%, the mean Hb increased from 7. 8 g/dl to 9.2 g/dl, and the mean ferritin level from 200 to 395 mg/dl. The mean EPO dosage could be tapered from 6,500 IU to 6,150 IU. No adverse side-effects were noted. Hence, in this uncontrolled study intravenous iron was an effective treatment for iron deficiency during rHuEPO therapy in children with ESRD on hemodialysis. PMID- 10460506 TI - Treatment of hyperhomocysteinemia in children on dialysis by folic acid. AB - Adult patients with renal failure have a high total homocysteine concentration in plasma. Hyperhomocysteinemia is an independent risk factor for cardiovascular diseases. Folic acid lowers the homocysteine concentrations in plasma in hyperhomocysteinemia. Whether this results in a reduced risk for cardiovascular diseases remains to be proven by intervention studies. In the present study we investigated: (1) if homocysteine concentrations are elevated in the plasma of children with renal failure and (2) the influence of folic acid administration on the plasma homocysteine concentration. The plasma homocysteine concentration was measured in 21 children, 9 on hemodialysis and 12 on peritoneal dialysis, before and 4 weeks after treatment with 2.5 mg folic acid daily. Healthy children (234) constituted the control group. In controls the median homocysteine concentration was 9.1 micromol/l (range 4.3-20.0 micromol/l). The median plasma homocysteine concentration in patients before folic acid treatment was 20.0 micromol/l (Q1-Q3 13.7-26.0; Q, quartile). After 4 weeks of folic acid treatment the median plasma homocysteine concentration was 12.0 micromol/l [Q1-Q3 9.8-14.3 (P<0.0001 Wilcoxon signed rank test)]. There was no significant difference between hemodialysis and peritoneal dialysis patients. Children with renal failure treated with hemodialysis or peritoneal dialysis have elevated plasma homocysteine concentrations, but this is significantly reduced after administration of 2.5 mg folic acid daily for 4 weeks. It is suggested that folic acid be added to the treatment of children with renal failure, although a beneficial effect still has to be proven. The required dose needs further study. PMID- 10460507 TI - Basic fibroblast growth factor in HIV-associated hemolytic uremic syndrome. AB - Endothelial injury is the primary pathogenic event leading to the renal thrombotic microangiopathic lesions typical of the hemolytic uremic syndrome (HUS). Basic fibroblast growth factor (bFGF) is an angiogenic growth factor released by injured endothelial cells. In a previous study we have found a significant accumulation of bFGF in human immunodeficiency virus (HIV)-transgenic mice with renal disease. Here we investigated whether bFGF was accumulated in the circulation and kidneys of two children with HIV-associated HUS (HIV-HUS), and studied the mechanisms involved in this process. The plasma levels of bFGF in children with HIV-HUS (124+/-20 pg/ml) were increased compared with five children with HIV nephropathy (49+/-6 pg/ml) and twenty HIV-infected children without renal disease (26+/-4 pg/ml, P<0.001). Immunohistochemistry and receptor binding studies showed that bFGF was accumulated bound to heparan sulfate proteoglycans in renal glomeruli and interstitium surrounding renal tubules in HIV-HUS kidneys. Basic FGF stimulated the proliferation of mesangial and urinary renal tubular epithelial cells isolated from both patients. These findings support the hypothesis that bFGF and its low-affinity binding sites may play a relevant role in modulating the process of glomerular and renal tubular regeneration during the acute stages of HIV-HUS. A follow-up study in a larger sample population is required to confirm these results. PMID- 10460508 TI - Etiology and outcome of chronic renal failure in Indian children. AB - A prospective analysis of all new pediatric cases of chronic renal failure (CRF) was performed at our hospital over a 1-year period. The diagnosis of CRF was based on serum creatinine >2 mg/dl with supportive clinical, laboratory, and radiological findings. There were a total of 48 patients with CRF with a median age of 13 years (range 10 days to 16 years). The causes of CRF included glomerulonephritis (37.5%), obstruction and interstitial (52%), hereditary (6.3%), and undetermined (4.2%). Patients were symptomatic for a mean of 33.2 months (range 10 days to 11 years) at presentation. Eight patients (16.7%) had acute reversible deterioration of renal function at presentation. This was due to accelerated hypertension in 2, infection in 3, volume depletion in 2, and nonsteroidal antiinflammatory drugs in 1 patient. At presentation, 22 (46%) children had mild to moderate renal failure and 26 (54%) had end-stage renal disease. Twenty-one children (43. 7%) had associated illness at presentation. Mean follow-up was 22.9 weeks (range 2-126 weeks). At the end of the study period, 10 (21%) patients were on conservative treatment, 7 (14.6%) on maintenance dialysis, 8 (16.7%) patients had functioning allografts, 4 (8.3%) patients had died, and 19 (39.6%) opted against further therapy. We conclude that CRF in Indian children carries a poor prognosis due to late referral and the limited availability and high cost of renal replacement therapy. PMID- 10460509 TI - Glomerulonephritis without IgA deposits in a case of Henoch-Schonlein purpura. AB - A boy aged 3 years 8 months with Henoch-Schonlein purpura (HSP) developed significant proteinuria with hematuria 2 days after the appearance of purpura rash. Although thought to be purpura nephritis, a percutaneous renal biopsy revealed diffuse mesangial proliferative glomerulonephritis (MesPGN) without deposition of immunoglobulin A or complement. Since his urine screening test during a health check at the age of 3.5 years had been unremarkable, HSP might have played a role in the pathogenesis of his non-IgA MesPGN. To our knowledge, non-IgA MesPGN is an uncommon renal manifestation of HSP. PMID- 10460510 TI - Minimal change nephrotic syndrome: a possible complication of ehrlichiosis. AB - Ehrlichiae are rickettsial organisms recently shown to be human pathogens. Infections often cause fever, myalgia, and hematological abnormalities, and sometimes mild elevation in transaminases, creatinine, and urinary protein. We report a teenager with nephrotic syndrome from minimal change glomerulonephritis and serological evidence of ehrlichiosis. In the appropriate clinical setting, Ehrlichiae should be considered in the etiological assessment of patients with minimal change disease. PMID- 10460511 TI - Disseminated autoimmune disease during levamisole treatment of nephrotic syndrome. AB - Side effects such as cutaneous vasculitis, which occur during prolonged levamisole treatment, may discourage the utilization of the drug in relapsing nephrotic syndrome. We describe a child who developed disseminated vasculitis during prolonged treatment with levamisole. The acute phase was characterized by hepatosplenomegaly, hemolytic anemia, IgM anticardiolipin and p-antineutrophil cytoplasmic antibodies. One month after withdrawal of therapy all symptoms had disappeared and tests normalized. This case report, together with other reports on cutaneous vasculitis, suggest caution and close monitoring during prolonged levamisole therapy. PMID- 10460512 TI - Nephrotic syndrome at 5 months: no definitive treatment or complications for 12 years. AB - We describe a patient who developed nephrotic syndrome at 5 months, with extensive glomerular and tubular damage on biopsy. The patient was treated with diuretics and was asymptomatic for a decade despite unremitting proteinuria. A repeat biopsy at 13 years of age showed remarkable healing with histopathological features consistent with "minimal change" nephrotic syndrome. This patient illustrates a favorable clinical outcome, without specific treatment, of nephrotic syndrome of long duration. PMID- 10460513 TI - PHEX expression in parathyroid gland and parathyroid hormone dysregulation in X linked hypophosphatemia. AB - X-linked hypophosphatemia (XLH), a renal phosphate (Pi) wasting disorder with defective bone mineralization, is caused by mutations in the PHEX gene (a Pi regulating gene with homology to endopeptidases on the X chromosome). Parathyroid hormone (PTH) status in XLH has been controversial, with the prevailing belief that hyperparathyroidism develops in response to Pi therapy. We report a 5-year old girl with XLH (patient 1) who had significant hyperparathyroidism at presentation, prior to initiation of therapy. We examined her response to a single oral Pi dose, in combination with calcitriol, and demonstrated a rise in serum concentration of intact PTH, which peaked at 4 h and paralleled the rise in serum Pi concentration. We also present two other patients whose parathyroid glands were analyzed for PHEX mRNA expression following parathyroidectomy. Patient 2 had autonomous hyperparathyroidism associated with chronic renal insufficiency, and patient 3, with XLH, developed autonomous hyperparathyroidism after 8 years of therapy with Pi and calcitriol. Following parathyroidectomy, patient 3 exhibited an increase in both serum Pi concentration and renal Pi reabsorption. The abundance of PHEX mRNA, relative to beta-actin mRNA, in parathyroid glands from patients 2 and 3 was several-fold greater than that in human fetal calvaria, as estimated by ribonuclease protection assay. In summary, we have shown that hyperparathyroidism can be a primary manifestation of XLH and that PHEX is abundantly expressed in the parathyroid gland. Given that PHEX has homology to endopeptidases, we propose that PHEX may have a role in the normal regulation of PTH. PMID- 10460514 TI - Molecular and cellular pathophysiology of obstructive nephropathy. AB - Congenital obstructive nephropathy remains one of the most-important causes of renal insufficiency in children. This review focuses on the unique interactions that result from urinary tract obstruction during the period of renal development in the neonatal rodent. Following unilateral ureteral obstruction (UUO), growth of the obstructed kidney is impaired and compensatory growth by the intact opposite kidney is related directly to the duration of obstruction. Development of the renal vasculature is delayed by UUO, and the activity of the intrarenal renin-angiotensin system is enhanced throughout the period of obstruction. Glomerular maturation is also delayed by UUO, and nephrogenesis is permanently impaired. The effects of UUO on the developing tubule are also profound, with a suppression of proliferation, stimulation of apoptosis, and the maintenance of an immature phenotype by tubular epithelial cells. Expression of tubular epidermal growth factor is suppressed and transforming growth factor-beta1 and clusterin are increased. Maturation of interstitial fibroblasts is delayed, with progression of tubular atrophy and interstitial fibrosis resulting in part from continued activation of the renin-angiotensin system and oxygen radicals. Future efforts to prevent the consequences of congenital urinary tract obstruction must account for the dual effects of obstruction: interference with normal renal development and progression of irreversible tubulointerstitial injury. PMID- 10460515 TI - The Wilms tumour gene, WT1, in normal and abnormal nephrogenesis. AB - The Wilms tumour gene, WT1, has been shown to play an important role in normal development of the kidney and gonad. Constitutional mutations predispose to both malformation and childhood tumours of these organs. There is a genotype-phenotype correlation, with missense mutations producing more severe abnormalities than complete absence of one allele. Two syndromes with early-onset protein-losing nephropathy can be distinguished according to the type of WT1 mutation. Children with apparently isolated diffuse mesangial sclerosis may also be WT1 mutation carriers. WT1 is not the major gene mutated in Wilms tumour, but has given important insights into the molecular genetics of this childhood embryonal kidney cancer. Recommendations for management of children suspected of having a WT1 mutation are discussed. PMID- 10460522 TI - Anaesthesia and the law: a philosophy of change. PMID- 10460523 TI - ICU admission in patients infected with the human immunodeficiency virus - a multicentre survey. AB - We conducted a retrospective study to assess the reasons for admission to the intensive care unit, and subsequent outcome, in patients infected with the human immunodeficiency virus (HIV). Four hospitals in the south of England participated, all with specialist HIV units. Data were collected on 127 patients admitted to ICU on 133 separate occasions between June 1993 and October 1997. The mean age on admission was 38 years (range 23-60 years). Ninety-four patients (70.7%) were documented HIV-positive before admission and 36 (27%) were diagnosed HIV-positive for the first time during admission; 36.1% were admitted with Pneumocystis carinii pneumonia. Overall ICU mortality was 33%, in-hospital mortality was 56% and the eventual mortality at the end of follow-up (March 1998) was 72%. Survival was highest in those admitted with respiratory HIV-related disease or HIV-unrelated illness. Associations with poor outcome included a prior AIDS-defining illness, a CD4 cell count of less than 100 cells.ml-1 and admission secondary to sepsis. PMID- 10460524 TI - The effect of anaesthesia and surgery on plasma cytokine production. AB - The aim of this study was to investigate cytokine production in response to anaesthesia [total intravenous anaesthesia (TIVA) with propofol, sufentanil and atracurium] and surgery (laparoscopic vs. open cholecystectomy). Forty adult patients, ASA I-II, undergoing elective laparoscopic (group 1) or open (group 2) cholecystectomy were studied. Venous blood samples for measurement of interleukin (IL)-1beta, IL-2, IL-4, IL-6, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) were taken before the induction of anaesthesia, pre incisionaly, at the end of anaesthesia and surgery and 24-h postoperatively. Pre incisionaly, in both groups, IL-1beta, IL-4, IL-6, TNF-alpha and IFN-gamma did not show a significant change, whereas IL-2 showed a significant decrease (p < 0.005 in group 1 and p < 0.001 in group 2) compared with pre-induction levels. By the end of anaesthesia and surgery, IL-1beta, IL-2, IL-4, IL-6 and TNF-alpha showed a significant increase in group 2 (p < 0.005 for IL-1beta, IL-2 and IL-4, and p < 0.05 for IL-6 and TNF-alpha); while in group 1, only IL-2 showed a significant increase (p < 0.01) and IFN-gamma showed a significant decrease (p < 0.05) compared with pre-incisional levels. By 24-h postoperatively, IL-1beta, IL 4, IL-6 and TNF-alpha had decreased significantly in group 2 (p < 0.005 for IL-4 and p < 0.05 for the others); whereas in group 1, IL-2 and IFN-gamma showed a significant increase (p < 0.005) compared with the end of anaesthesia and surgery level. In conclusion, TIVA with propofol, sufentanil and atracurium does not seem to have a significant effect on IL-1beta, IL-4, IL-6, TNF-alpha and IFN-gamma release. IL-2 was the only cytokine to show a significant decrease due to the effect of anaesthesia alone in both groups. The cytokine response to open cholecystectomy stimulated both the pro-inflammatory (IL-1beta, IL-6 and TNF alpha) and the anti-inflammatory (IL-4) components, while this response was absent in laparoscopic cholecystectomy. PMID- 10460525 TI - Prognostic indicators following emergency aortic aneurysm repair. AB - We performed a retrospective study of 135 patients presenting for emergency abdominal aneurysm repair to determine predictive factors for outcome. The outcome measures investigated were mortality in the operating theatre and intensive care, and at 28 and 100 days. Univariate analysis showed that the patient's age, hypotension on admission, aneurysmal rupture, pre-operative cardiopulmonary resuscitation, intra-operative blood loss and hypotension were risk factors for death either in the operating theatre or up to 100 days after surgery. Binary logistic regression identified the independent risk factors for survival. Operative survival was determined by acute factors such as pre operative cardiopulmonary resuscitation, aneurysmal rupture and intra-operative hypotension. Longer term survival was determined by the patient's age, aneurysmal rupture, blood loss and blood pressure at admission. Using a binary logistic regression equation, from which a simplified risk score was derived, it is possible to predict the likelihood of survival of individual patients presenting for abdominal aortic aneurysm repair. PMID- 10460526 TI - Target-controlled propofol vs. sevoflurane: a double-blind, randomised comparison in day-case anaesthesia. AB - We compared target-controlled propofol with sevoflurane in a randomised, double blind study in 61 day-case patients. Anaesthesia was induced with a propofol target of 8 microgram.ml-1 or 8% sevoflurane, reduced to 4 microgram.ml-1 and 3%, respectively, after laryngeal mask insertion and subsequently titrated to clinical signs. Mean (SD) times to unconsciousness and laryngeal mask insertion were significantly shorter with propofol [50 (9) s and 116 (33) s, respectively] than with sevoflurane [73 (14) s and 146 (29) s; p < 0.0001 and p = 0.0003, respectively]; however, these differences were not apparent to the blinded observer. Propofol was associated with a higher incidence of intra-operative movement (55 vs. 10%; p = 0.0003), necessitating more adjustments to the delivered anaesthetic. Emergence was faster after sevoflurane [5.3 (2.2) min vs. 7.1 (3.7) min; p = 0.027], but the inhaled anaesthetic was associated with more nausea and vomiting (30 vs. 3%; p = 0.006), which delayed discharge [258 (102) min vs. 193 (68) min; p = 0.005]. Direct costs were lower with sevoflurane but nausea would have increased indirect costs. Patient satisfaction was high (>/= 90%) with both techniques. In conclusion, both techniques had advantages and disadvantages for day-case anaesthesia. PMID- 10460527 TI - Anaesthesia clinical directors in the United Kingdom: organisation, objectives and support needs. AB - A postal survey of all 269 acute hospital trusts identified in the United Kingdom was carried out to study the work of Clinical Directors of anaesthesia. Initial responses from 163 Clinical Directors and 129 completed questionnaires were analysed. Four main areas of concern revealed by the survey were contracts and objectives, funding of managerial sessions, access to information and perceived need for support. Most Clinical Directors had no job description and most had no formal written objectives, despite a substantial body of advice that these should be provided. There was generally substantial underfunding of managerial hours compared with those actually worked and approximately 20% of Clinical Directors surveyed had no funding for managerial duties. Clinical Directors' ratings of the information available to assist their decision making were also a cause of concern. Clinical Directors perceived that they need better networking, more training particularly on human resource management and improved management information. PMID- 10460528 TI - Provision of training in chronic pain management for specialist registrars in the United Kingdom. AB - A study published in 1992 highlighted wide variations in the provision of training in pain management. In this survey, data were collected from both pain clinicians and Programme Directors of the Schools of Anaesthesia to see if there had been any changes in training patterns since the introduction of the Calman training scheme. There did not seem to be a uniform improvement in the provision of training in pain management for Specialist Registrars and many may reach their Certificate of Completion of Specialist Training without a basic knowledge of chronic pain. It is thought that at the present time there will be few Specialist Registrars with sufficient training to take up consultant posts in pain management unless they compete for the much sought after, and often not fully funded, pain fellowships outside their rotations. PMID- 10460529 TI - Hemispheric-synchronisation during anaesthesia: a double-blind randomised trial using audiotapes for intra-operative nociception control. AB - The possible antinociceptive effect of hemispheric-synchronised sounds, classical music and blank tape were investigated in patients undergoing surgery under general anaesthesia. The study was performed on 76 patients, ASA 1 or 2, aged 18 75 years using a double-blind randomised design. Each of the three tapes was allocated to the patients according to a computer-generated random number table. General anaesthesia was standardised and consisted of propofol, nitrous oxide 66%/oxygen 33%, isoflurane and fentanyl. Patients breathed spontaneously through a laryngeal mask and the end-tidal isoflurane concentration was maintained near to its minimum alveolar concentration value of 1.2%. Fentanyl was given intravenously sufficient to keep the intra-operative heart rate and arterial blood pressure within 20% of pre-operative baseline values and the fentanyl requirements were used as a measure of nociception control. Patients to whom hemispheric-synchronised sounds were played under general anaesthesia required significantly less fentanyl compared with patients listening to classical music or blank tape (mean values: 28 microgram, 124 microgram and 126 microg, respectively) (p < 0.001). This difference remained significant when regression analysis was used to control for the effects of age and sex. PMID- 10460530 TI - Effects of lamotrigine on pain-induced chemo-somatosensory evoked potentials. AB - Lamotrigine, a sodium channel blocker that selectively inhibits the neuronal release of glutamate, has been shown to produce analgesia in acute and chronic pain models in rats without causing noticeable sedation. After oral administration it also reduces pain scores, as assessed by the cold pain test, in volunteers. The purpose of this study was to determine the analgesic effect of lamotrigine given by mouth to healthy volunteers as evidenced by alterations in chemo-somatosensory evoked potentials. The following factors were measured: latency to N1 and P100 peak (ms); amplitude between the N1 and P100 peak (microV); visual analogue pain intensity scores. A double-blind, randomised and crossover design was used in which 12 volunteers received either placebo or lamotrigine 300 mg on separate occasions as determined by the randomisation schedule. Volunteers were tested before and 2 h after the treatment. The plasma lamotrigine concentration was measured immediately after the end of the experimental sessions. Lamotrigine produced a significantly higher latency to P100 values at 2 h postdrug than placebo (p < 0.05) but had no significant effects on the other factors. Although plasma concentrations were similar to those observed in the cold pain test, we conclude that lamotrigine 300 mg by mouth had no analgesic effect in this acute pain model. PMID- 10460531 TI - The 'dedicated airway': a review of the concept and an update of current practice. AB - The term 'dedicated airway' was first used in connection with nasal fibreoptic intubations using the cuffed nasopharyngeal airway. Since that time, the concept has developed and the term has been extended to include fibreoptic intubation techniques involving both the laryngeal mask airway and cuffed oropharyngeal airway. 'Dedicated airway' can now be defined as: 'An upper airway device dedicated to the maintenance of airway patency while other major airway interventions are anticipated or are in progress. The device should be compatible with spontaneous and controlled ventilation. 'Dedicated airway techniques allow planned fibreoptic intubations in difficult cases and provide an emergency airway option in an unexpected difficult intubation when the alternative may be to wake the patient. As well as promoting safe conditions for training fibreoptic intubation in general, there is the particular advantage of being able to train using these techniques in patients known to be difficult to intubate. The authors' evolved clinical experiences in promoting the concept and the relevant literature are reviewed. PMID- 10460532 TI - Acromegaly and papillomatosis: difficult intubation and use of the airway exchange catheter. AB - We describe the anaesthetic management of a patient with acromegaly scheduled for transsphenoidal resection of a pituitary tumour who was found at intubation to have coexisting laryngeal papillomatosis. Oral intubation was impossible using both direct and fibreoptic techniques. Nasal fibreoptic intubation was successful but precluded the transsphenoidal approach to surgery. A Cook Airway Exchange Catheter [Cook (UK) Ltd, Monroe House, Letchworth SG6 1LN] was used with a Negus bronchoscope to convert to oral intubation and allow completion of surgery without resort to tracheostomy. PMID- 10460533 TI - Air emboli with Haemaccel(R) AB - We report two cases of venous air embolism which occurred in association with infusion of Haemaccel(R) using a pressure bag. As a result of these incidents, we performed a study that showed that up to 45 ml of air can be infused into a patient from a pressurised Haemaccel(R) plastic bottle using a standard administration set. We also demonstrated that the volume of air infused was influenced by the type and size of the pressure bag and the warming of the Haemaccel(R) plastic container. PMID- 10460534 TI - Intubating laryngeal mask and rapid sequence induction in patients with cervical spine injury. AB - The Intubating Laryngeal Mask (FastrachTM), a modified conventional laryngeal mask airway, and its prototype cuffed silicone tube, continue to be an appropriate intubating tool in combination with fibreoptic bronchoscopy in the emergency situation. This is an account of two patients with suspected cervical spine fracture admitted to our emergency room in a haemodynamically unstable condition and requiring a rapid sequence induction of anaesthesia, in whom we successfully applied this newly developed intubating device for the first time. Provided that there are no intubation difficulties, direct laryngoscopy is still the fastest method of securing an airway; however, this procedure leads to an extension of the cervical spine, which may be hazardous in the case of a cervical spine injury. Intubation by means of the Intubating Laryngeal Mask avoids dangerous hyperextension of the occipito-atlanto-axial complex, a fact that we were able to verify by lateral cervical spine fluoroscopy during intubation. PMID- 10460535 TI - Effect of cricoid pressure on gastro-oesophageal reflux in awake subjects. AB - This study aimed to evaluate whether cricoid pressure is associated with a high risk of gastro-oesophageal reflux. Fifteen awake, fasted volunteers were studied. A cricoid pressure of 44 N was applied for 60 s by resting a padded yoke over the cricoid cartilage. Using continuous oesophageal pH monitoring, no volunteer had gastro-oesophageal reflux during cricoid pressure, although one subject had a reflux spike soon after relieving cricoid pressure. We conclude with 95% confidence that the incidence of gastro-oesophageal reflux during cricoid pressure is not more than 20%. PMID- 10460536 TI - Smokers and haemodynamic responses to desflurane. AB - Changes in basic haemodynamic variables following a sudden increase in the concentration of inspired desflurane administered during anaesthesia were compared in 30 female smokers and 30 nonsmokers. Smokers had significantly greater percentage increases in heart rate (p = 0.02), systolic and diastolic blood pressure (p < 0.005 and = 0.01, respectively) and rate-pressure product (p < 0.005). Such increases were reached faster by smokers for both systolic and diastolic blood pressures (p < 0.01) and for rate-pressure product (p = 0.02). The peak values of rate pressure product reached were sufficient to warrant caution when using desflurane for anaesthesia in smokers. PMID- 10460537 TI - Long-term outcome of percutaneous thermocoagulation for trigeminal neuralgia. AB - A retrospective analysis of long-term efficacy of percutaneous radiofrequency thermocoagulation of the trigeminal ganglion or root for the relief of trigeminal neuralgia was carried out in our unit. From the medical records and questionnaires, outcomes of 108 procedures performed in 81 patients from January 1986 to December 1990 were obtained with a follow-up period of 6-11 years. The initial success rate was 87% and the probability of remaining pain-free 1, 2 and 11 years after the procedure was 65, 49 and 26%, respectively. Patients with typical symptoms had a better long-term efficacy than those with atypical presentations, and patients who had not undergone a previous surgical procedure also had a better outcome. There was no mortality in this series. Common adverse effects included dysaesthesia in 20 patients, corneal numbness in 12 patients and masseter weakness in three patients. PMID- 10460538 TI - Paediatric intensive care - specialisation reduces mortality. PMID- 10460539 TI - Paediatric intensive care - specialisation reduces mortality PMID- 10460540 TI - Training method for placement of the laryngeal mask. PMID- 10460541 TI - Airway difficulties associated with severe epistaxis. PMID- 10460542 TI - Emergency tracheostomy tube change: another use of the tracheal tube. PMID- 10460543 TI - Tonsillectomy - an unusual complication. PMID- 10460544 TI - A potential complication of caudal anaesthesia. PMID- 10460545 TI - Peri-operative silent myocardial ischaemia. PMID- 10460546 TI - Co-induction for ambulatory surgery. PMID- 10460547 TI - Waiting for generic propofol. PMID- 10460548 TI - Ethical approval for research in anaesthesia. PMID- 10460549 TI - An unfortunate allergy. PMID- 10460550 TI - 'Mighty things from small beginnings grow' John Dryden (1631-1700) Annus mirabilis. PMID- 10460551 TI - Evaluation of self-reported failures in cognitive function after cardiac and noncardiac surgery. AB - Long-term cognitive deficits after cardiac surgical procedures involving cardiopulmonary bypass have been well documented. The occurrence of prolonged cognitive changes after noncardiac surgery has not, however, been clearly established. Using the Cognitive Failures Questionnaire, which permits self assessment of cognitive impairment, we studied 50 patients before and 2 months after coronary bypass surgery and major vascular surgical procedures. Pre operative test scores did not differ between groups. Postoperatively, 24 cardiac surgical patients and 22 vascular surgical patients completed the questionnaire. Both groups reported significantly more cognitive failures occurring after surgery than in the pre-operative period. This suggests that there are factors other than the exposure to cardiopulmonary bypass during cardiac surgery that affect self-assessed, long-term postoperative cognitive sequelae. PMID- 10460552 TI - The cardiorespiratory effects of laparoscopic procedures in infants. AB - We assessed the cardiorespiratory effects of laparoscopic procedures in 27 infants aged between 36 and 365 days. Infants were monitored and anaesthetised in a standardised manner. Heart rate, mean arterial pressure, end-tidal carbon dioxide and oxygen saturation were recorded, and blood gases were measured at 5 min after intubation, 15 and 30 min after carbon dioxide pneumoperitoneum, 5 min after desufflation and after extubation. The pH, PaO2, base excess, SaO2 and SpO2 decreased, and PCO2 increased by insufflation of carbon dioxide intraperitoneally, and improved following deflation. Changes in pH and PaO2 during the study were statistically significant (p < 0.0001). The increase in PaCO2 30 min after pneumoperitoneum was statistically significant when compared with initial values. Transient arrhythmias were observed in 10 infants 1 min after pneumoperitoneum. There were no statistically significant alterations in heart rate and systolic blood pressure. PMID- 10460553 TI - Time course of changes in breathing pattern in morphine- and oxycodone-induced respiratory depression. AB - The time course of changes in breathing pattern in opioid-induced respiratory depression was characterised for two opioids. Intravenous morphine (0.039 mg.kg-1 bolus + 0.215 mg.kg-1.h-1 infusion) and oxycodone (0.05 mg.kg-1 bolus + 0.275 mg.kg-1.h-1 infusion) were administered to six healthy male volunteers for 2 h in a random, double-blind and cross-over fashion. Monitoring included pulse oximetry and noninvasive respiratory-inductive plethysmography for the measurement of breathing pattern. The total amounts of drugs given were 35.1 (0.0) mg [mean (SD)] morphine and 41.3 (8.0) mg oxycodone. Four of the six oxycodone infusions had to be stopped at 99 (14) min because of respiratory depression as judged by pulse oximetry. No morphine infusions were stopped. The first changes in breathing pattern were a decrease in respiratory rate and an increase in the contribution of the rib cage to tidal volume, while the compensatory increase in tidal volume became evident later. A decrease in minute ventilation and inspiratory duty cycle were also found. PMID- 10460554 TI - Sevoflurane: a comparison between vital capacity and tidal breathing techniques for the induction of anaesthesia and laryngeal mask airway placement. AB - Sixty unpremedicated adult day-case patients were randomly assigned to either vital capacity or tidal breathing inhalational induction techniques. End points assessed included loss of eyelash reflex, time to drop a weighted syringe, time to jaw relaxation and time to the end of laryngeal mask airway insertion. Complications occurring during the induction of anaesthesia were recorded. The data show that there is no statistical or clinical difference between the two induction techniques. Patient acceptance of both techniques was similarly high. When the time taken to prime the anaesthetic breathing system is taken into consideration, the vital capacity technique is more expensive for induction of anaesthesia. These results therefore question the need for the vital capacity induction technique with sevoflurane 8%. PMID- 10460555 TI - Critically ill medical patients, their demographics and outcome. AB - There are few reports describing the demographic details and outcome of noncoronary medical patients on adult general intensive care units. It is not known how medical patients differ from other critically ill patients and how this may influence their outcome. Consequently, we recorded the demographic details of 374 critically ill medical patients and followed their survival for up to 3 years. Patients referred from medical specialties are younger, more severely ill and suffer a higher severity-of-illness-adjusted intensive care unit mortality than other patients. The short-term survival of medical patients is poor with a median survival of 40 days. Twenty per cent of medical patients die after discharge from intensive care but before 40 days. However, the long-term survival of medical patients is better than other patients and almost as good as the general population. Further research is required to identify those patients who are likely to survive beyond 40 days. PMID- 10460556 TI - The patient-at-risk team: identifying and managing seriously ill ward patients. AB - A 'patient-at-risk team', established to allow the early identification of seriously ill patients on hospital wards, made 69 assessments on 63 patients over 6 months. Predefined physiological criteria were not able to reliably predict which patients would be admitted to the intensive care unit. The incidence of cardiopulmonary resuscitation before intensive care admission was 3.6% for patients seen by the team and 30.4% for those not seen (p < 0.005). Of admissions seen by the team, 25% died on the intensive care unit compared with 45% of those not seen (not significant, p = 0.07). Among those not seen by the team, mortality was 40% for those who did not require resuscitation and 57% for those who did (not significant). Many critically ill ward patients had abnormal physiological values before intensive care unit admission. Identification of critically ill patients on the ward and early advice and active management are likely to prevent the need for cardiopulmonary resuscitation and to improve outcome. PMID- 10460557 TI - Adrenocortical function in critically ill patients 24 h after a single dose of etomidate. AB - We compared the effects of single doses of etomidate and thiopentone on adrenocortical function in a randomised controlled clinical trial involving 35 critically ill patients who needed a general anaesthetic. Just before induction of anaesthesia, a baseline blood cortisol sample was taken. Twenty-four hours later we performed a short adrenocorticotrophic hormone stimulation test. No patient had a low cortisol level (< 160 nmol.l-1) at any time during the study. Baseline, pre-ACTH and post-ACTH cortisol levels were similar in the two groups. However, significantly more patients in the etomidate group had an ACTH stimulated cortisol increment < 200 nmol.l-1. The clinical significance of these findings is not clear, but we conclude that single doses of etomidate may interfere with cortisol synthesis for at least 24 h in the critically ill. PMID- 10460558 TI - Antiseptic-bonded central venous catheters and bacterial colonisation. AB - This study was undertaken to evaluate the impact of chlorhexidine/silver sulphadiazine-bonded catheters on the incidence of colonisation and catheter related sepsis in critically ill patients. Threehundred and fifty-one catheters were inserted into 228 patients during the study period, 174chlorhexidine/silver sulphadiazine-bonded catheters and 177 standard catheters. Indications for catheter removal were: death, clinical redundancy and clinical evidence of local or systemic infection. All catheter tips were sent to the microbiology laboratory for semiquantitative analysis of bacterial colony count. Seventy-one (40.2%) of the standard catheters and 47 (27.2%) of the antiseptic-bonded catheters were found to be colonised on removal (p < 0.01). Eight cases (4.7%) of catheter related sepsis were associated with standard catheters and three cases (1.7%) with antiseptic-bonded catheters, however, this reduction was not statistically significant. Our results indicate that the use of antiseptic-bonded catheters in critically ill patients significantly reduces the incidence of bacterial colonisation. PMID- 10460559 TI - Fibre-optically lit laryngoscope. AB - The light from a battery-powered laryngoscope with a fibreoptic blade is limited by the 3-V battery and deteriorates with use, as the optic fibres break. A laryngoscope handle is described in which the bulb is replaced by a fibreoptic cable connecting to a mains, halogen light source. This laryngoscope gives 6000 Cd.m(-2) light at the centre of the field compared with 800 Cd.m(-2) for the battery. The field of illumination is also improved from 1600 Cd.m(-2) 20 mm from the centre compared with 120 Cd.m(-2) for the battery handle. The fibre optic cable is attached at the side of the handle and does not interfere with intubation. The mains-powered light source has the advantages of being reliable with the ability to switch immediately to a second bulb; low running costs; improved light for intubation and it makes it possible to use a dental mirror for indirect laryngoscopy in difficult intubations. PMID- 10460560 TI - The effect of steam sterilisation at 134 degrees C on light intensity provided by fibrelight Macintosh laryngoscopes. AB - We studied the repeated effect of sterilisation on light intensity in laryngoscopes from Penlon, Riester, Heine (two different blades), Medicon and Upsher. Light intensity was measured by a light meter using two methods. Measurements were performed before the decontamination procedure was carried out and subsequently after each series of 25 procedures until a total of 200 cycles was reached. Using method 1 (and 2), the reduction in light intensity after 200 cycles was 100% (100%; no light emitted), 37% (13%), 75% (69%), 79% (60%), 37% (14%) and 63% (55%) for each blade, respectively. PMID- 10460561 TI - The use of neuromuscular blocking agents in noncardiac surgery after dynamic cardiomyoplasty. AB - Dynamic cardiomyoplasty is a surgical treatment to improve cardiac performance in patients with end-stage heart failure by wrapping the latissimus dorsi muscle around the heart. The use of skeletal muscle raises concerns about the safety of neuromuscular blocking agents used during general anaesthesia in noncardiac surgery in patients after cardiomyoplasty. We describe the administration of rocuronium to a patient undergoing carotid endarterectomy 18 months after cardiomyoplasty. No clinically relevant effects on haemodynamics were observed. We conclude that the use of nondepolarising neuromuscular blocking agents for noncardiac surgery in patients after cardiomyoplasty does not compromise cardiac performance in a clinically relevant way, although the time between the cardiomyoplasty procedure and the use of nondepolarising neuromuscular blocking agents remains a concern. PMID- 10460562 TI - Exaggerated cardiovascular response to anaesthesia--a case for investigation. AB - We present a case of a 40-year-old woman who developed major cardiovascular complications during anaesthesia for an elective clipping of a cerebral arteriovenous malformation. Postoperative investigation confirmed the diagnosis of an adrenal phaeochromocytoma. In retrospect, it became apparent that she had experienced a series of potentially life-threatening events over a 20-year period all of which are known complications of phaeochromocytoma. This case highlights the importance of investigating young patients who have unexpected and unexplained cardiovascular events during anaesthesia and surgery. PMID- 10460563 TI - Failure of insertion of a laryngeal mask airway caused by a variation in the anatomy of the thyroid cartilage. AB - We report the failure of insertion of a laryngeal mask airway in a patient with a pre-operative diagnosis of an abnormality of the superior cornua of the thyroid cartilage. We believe that this is the first time that this reason for failure has been reported. PMID- 10460564 TI - Anaesthetic management of Caesarean section in an elderly parturient with pre eclampsia. AB - The number of women over 40 years of age becoming pregnant has increased over recent years. They suffer a high incidence of hypertensive complications, and require more frequent operative interventions. We present a case report of a 51 year-old woman having a Caesarean section for a twin pregnancy complicated by pre eclampsia. We discuss the effects of age on pregnancy and the implications for anaesthetic management. PMID- 10460565 TI - Hyperbaric oxygen therapy in the management of severe acute anaemia in a Jehovah's witness. AB - A case is described in which a Jehovah's Witness patient who refused blood transfusion suffered massive antepartum haemorrhage, her haemoglobin falling as low as 2.0 g.dl(-1). She was treated on an intensive care unit with intermittent positive pressure ventilation and general supportive measures, pulsed hyperbaric oxygen therapy and recombinant human erythropoietin. PMID- 10460566 TI - Profound motor blockade with epidural ropivacaine following spinal bupivacaine. AB - Ropivacaine, a relatively new amide local anaesthetic, reputedly produces less motor block than equivalent doses of bupivacaine, potentially combining high quality analgesia with the ability to ambulate. We report two cases of prolonged, profound motor block with patient-controlled epidural analgesia using 0.1% ropivacaine, following spinal bupivacaine for Caesarean section. As there was no evidence of inadvertent intrathecal ropivacaine administration or of any neurological injury, we hypothesise that epidural ropivacaine may interact with intrathecal bupivacaine to prolong its effect. PMID- 10460567 TI - Effect of epidural bupivacaine on the relationship between the bispectral index and end-expiratory concentrations of desflurane. AB - We compared the relationship between the bispectral index and end-tidal desflurane concentrations in 20 patients undergoing elective surgery. Patients received epidurally either 10 ml saline (group S) or 10 ml bupivacaine 0.125% with epinephrine 1/800 000 (group B) before induction of anaesthesia with sufentanil (0.15 microgram.kg(-1)) and propofol (2 mg.kg(-1)); muscle relaxation was obtained with cisatracurium (0.2 mg.kg(-1)). Patients lungs were ventilated to maintain end-tidal desflurane at 3% in O2/N2O (50/50) until 5 min after skin incision, followed by two consecutive 10 min periods at end-tidal desflurane 6% and 9%. bispectral index values were recorded before induction, at 3% desflurane before and 5 min after skin incision, and at 6% and 9% end-tidal desflurane. Bispectral index decreased with increasing end-tidal desflurane concentration (ANOVA: p < 0.05). The decrease in bispectral index was significant between pre induction, 3% and 6% desflurane. No significant difference was observed at 3% desflurane before and after skin incision, or between 6 and 9% desflurane. The relationship between bispectral index and end-tidal desflurane concentration was fitted by a linear regression in each group. No significant difference in bispectral index was observed between the groups at any time. We conclude that bispectral index decreases with increasing desflurane concentration and that this relationship is not affected by epidural 0.125% bupivacaine. PMID- 10460568 TI - The effects of ropivacaine hydrochloride on coagulation and fibrinolysis. An assessment using thromboelastography. AB - Amide local anaesthetics impair coagulation by inhibition of platelet function and enhanced fibrinolysis. The potential therefore exists that the presence of amide local anaesthetics in the epidural space could contribute to the therapeutic failure of an epidural autologous blood patch. Ropivacaine is an aminoamide local anaesthetic increasingly used for epidural analgesia and anaesthesia, particularly in obstetric practice. This study was undertaken to investigate whether concentrations of ropivacaine in blood, which could occur clinically in the epidural space, alter coagulation or fibrinolysis. Thromboelastography was used to assess clotting and fibrinolysis of blood to which ropivacaine had been added. Although modest alterations in maximum amplitude, coagulation time and alpha angle were observed, the effect of ropivacaine on clotting and fibrinolysis was not clinically significant. We conclude that it is unlikely that the presence of ropivacaine in the epidural space would reduce the efficacy of an early or prophylactic epidural blood patch. PMID- 10460569 TI - The pre-operative ECG in day surgery: a habit? AB - As the population presenting for day-case surgery and anaesthesia increases, so does the challenge of adequate pre-operative assessment. Although an electrocardiogram is frequently performed, its value in day-case surgery remains unproven. One thousand, one hundred and eighty-five patients presenting for day case surgery were assessed. One hundred and fifty-four (13%) were referred for electrocardiogram according to well-recognised criteria for the prediction of coronary artery disease. They were read independently by the anaesthetist responsible for the case and by an experienced cardiologist. A significant abnormality was noted in 26% of electrocardiograms, most frequently in patients referred with hypertension. There was a good correlation between the reports of the anaesthetist and cardiologist. Only 20% of those patients with an abnormal electrocardiogram had their surgery postponed. No adverse events occurred in patients proceeding to surgery despite the abnormalities. We conclude that a resting electrocardiogram is of limited value in risk stratification of patients undergoing day-case surgery. PMID- 10460570 TI - Clinical governance--what's it all about? PMID- 10460571 TI - Neck flexion and the intubating laryngeal mask. PMID- 10460573 TI - Airway problems in the recovery room. PMID- 10460574 TI - Nausea and vomiting during caesarean section under spinal anaesthesia. PMID- 10460575 TI - Epidural catheter clamp fixes too firmly. PMID- 10460576 TI - Painful ear sign. PMID- 10460578 TI - Why not just ask the parent? PMID- 10460577 TI - Problems involved with anaesthetising patients with DHPR deficiency. PMID- 10460579 TI - Experts in our field? PMID- 10460580 TI - Undecipherable abbreviations. PMID- 10460581 TI - Demonstration of an X chromosome in a neutrophil drumstick. PMID- 10460582 TI - Is albumin harmful? PMID- 10460583 TI - Iron deficiency in children: detection and prevention. PMID- 10460584 TI - Time trends in incidence of haematological malignancies and related conditions. PMID- 10460585 TI - The AML1-ETO chimaeric transcription factor in acute myeloid leukaemia: biology and clinical significance. PMID- 10460586 TI - The role of adoptive immunotherapy in the prevention and treatment of lymphoproliferative disease following transplantation. PMID- 10460587 TI - Haemopoietic reconstitution after sublethal irradiation: comparison of the effects of different haemopoietic cytokines on murine lymphocytes and bone marrow cells. AB - Sublethally irradiated mice were injected with recombinant cytokines to stimulate haemopoietic reconstitution. Interleukin (IL)-11 and IL-6 were able to significantly accelerate the recovery of thymus, spleen and bone marrow cells when used in combination with IL-3, but not alone. Stem cell factor (SCF) also displayed detectable effects when used with IL-3. Conversely, the IL-6 superagonist K-7/D-6 was able, even when used alone, to induce recovery of thymus, spleen and bone marrow cells up to the level of unirradiated controls. Together, these results indicate that it is possible to attain complete recovery of lymphoid organs and tissues as early as 7 d after irradiation by use of haemopoietic cytokines. PMID- 10460588 TI - Erythroblasts from friend virus infected- and phenylhydrazine-treated mice accurately model erythroid differentiation. AB - The dynamics of gene expression during terminal erythroid differentiation have been examined in three murine models; the erythroleukaemia cell line HCD-57 and splenic erythroblasts isolated from mice treated with either the anaemia-inducing strain of Friend virus (FVA cells) or the haemolytic agent phenylhydrazine (PHZ cells). In response to erythropoietin (EPO) and haemin, HCD-57 cells proliferated and synthesized haemoglobin, but failed to complete terminal differentiation as indicated by lack of change in both gene expression and morphological appearance. In contrast, EPO-induced terminal differentiation in FVA and PHZ cells in vitro was accompanied by increases in haemoglobin positivity, morphological maturation and a shared pattern of gene expression. EPO receptor (EPO-R) mRNA levels peaked before globin gene expression which was maximal at 24 h. Peak GATA-1 and EKLF mRNA levels also preceded the globin gene peak, but the highest NF-E2 levels coincided with maximal globin levels, suggesting a role for NF-E2 in the maintenance, rather than the initiation of globin gene expression. Peak expression of delta-aminolaevulinic acid synthase (ALAS) coincided with peak globin expression. FVA and PHZ cells represent more effective models than the HCD 57 cell line for the investigation of erythroid gene expression during EPO regulated terminal erythropoiesis. PMID- 10460589 TI - Def-2, -3, -6 and -8, novel mouse genes differentially expressed in the haemopoietic system. AB - To identify developmentally regulated genes during myeloid differentiation, a self-inactivating retroviral gene-trap vector carrying a beta-galactosidase neomycin (SA/lacZ/neo) fusion gene was constructed and used to infect myeloid progenitor cells (FDCP-Mix A4). G418-resistant and beta-galactosidase positive cell lines (gene-trap integration [GTI] clones) were established and induced to differentiate in vitro into either macrophages or granulocytes. Expression of the trapped loci was monitored at a single-cell level by analysing the mature cell types for beta-galactosidase activity. All 37 GTI clones tested showed down regulation either during granulocyte or both granulocytic and macrophage differentiation. The endogenous coding regions fused to the SA/lacZ/neo reporter gene were isolated from eight clones. Molecular analysis revealed that half of them represented novel mouse genes (def-2, -3, -6 and -8) which we confirmed to be differentially expressed in primary haemopoietic tissues. Database searches revealed no significant similarities for def-2 (associated with haemopoietic progenitors) and def-8 (expressed most strongly in peripheral leucocytes). Def-6, which is down-regulated upon the differentiation into myeloid as well as erythroid lineages, was found to be closely related but not identical with the recently described B-cell-specific switch recombinase SWAP-70. Def-3, which is down-regulated upon differentiation into granulocytes but expressed in progenitor cells and macrophages, defines a novel family of RNA binding proteins. PMID- 10460590 TI - Interaction of thrombopoietin with the platelet c-mpl receptor in plasma: binding, internalization, stability and pharmacokinetics. AB - Thrombopoietin (TPO) is the primary regulator of platelet production and acts through binding its receptor, c-mpl, found on megakaryocyte progenitor cells, megakaryocytes and platelets. Circulating levels of TPO are regulated primarily by the clearance of TPO after it binds to c-mpl receptors on circulating platelets. In this study the interaction of TPO with the platelet c-mpl receptor has been analysed under physiological conditions using radiochemical and pharmacokinetic approaches. 125I-rHuTPO was prepared using a novel method of gentle iodination that preserved its biological activity and used to demonstrate that platelets, but not endothelial cells, have a single class of binding sites (56 +/- 17 binding sites/platelet) with high affinity (Kd = 163 +/- 31 pM). Cross linking experiments confirmed that TPO, but not erythropoietin (EPO), specifically associated with the 95 kD platelet c-mpl receptor. Upon addition of TPO to platelets, 80% of the TPO binding sites were internalized within an hour and were not recycled. TPO that was not bound by platelets was stable for up to 6 d in both platelet-poor and platelet-rich plasma. Using unlabelled recombinant human TPO (rHuTPO), standard pharmacokinetic analysis demonstrated that platelets have an average TPO clearance of 1.24 +/- 0.38 ml/h/109 platelets and that TPO clearance was reduced by low temperature but not by a number of drugs or metabolic inhibitors. The maximal amount of TPO removed by platelets in vitro was identical to that predicted by the total number of TPO binding sites. These results provide a biochemical and pharmacokinetic basis for the clinical use of TPO and for understanding possible disorders of platelet production. PMID- 10460591 TI - Early progenitor cells from human mobilized peripheral blood express low levels of the flt3 receptor, but exhibit various biological responses to flt3-L. AB - The biological effects of flt3-L, and the expression of its tyrosine kinase receptor (flt3, CD135) were investigated on the immature subsets of human circulating peripheral blood progenitors obtained from cancer patients or normal volunteer donors, after mobilization with rhG-CSF or chemotherapy. flt3 was expressed at low levels, and its expression increased concomitantly with expression of CD38 within the CD34+ cell population. Despite this low-level expression, flt3-L exerted synergistic effects with a combination of c-kit ligand, IL-3, IL-6, GM-CSF and G-CSF, mainly to induce proliferation of CD34+/CD38- cells. In addition, flt3-L increased the detection of HPP-CFC, both immediately after cell selection, and after 7 and 14 d of cultures. We conclude that flt3-L is active on circulating early mobilized haemopoietic progenitors, despite the low- level expression of its receptor. PMID- 10460592 TI - Circadian variation of granulocyte colony stimulating factor levels in man. AB - Glucocorticoids dose-dependently increase plasma levels of granulocyte colony stimulating factor (G-CSF). Based on the marked circadian rhythm of cortisol levels, we hypothesized that plasma levels of G-CSF may also show a diurnal rhythm. A prospective study was conducted in 12 healthy young volunteers. Blood samples were obtained every 2 h over 24 h. G-CSF levels averaged 18.0 ng/l (CI 13. 1-22.9) at 8.00 am, increased continuously and reached peak values at 10.00 p.m. Individual harmonic regression analysis showed a clear circadian rhythm. The individual differences between nadir and peak levels averaged 54% (CI 43-65%). This pronounced diurnal rhythm of G-CSF levels may help understand the circadian changes in circulating stem cells, bone marrow DNA synthesis, or bone marrow toxicity induced by some chemotherapeutic agents. PMID- 10460593 TI - Selective effect of cyclosporine monotherapy for pure red cell aplasia not associated with granular lymphocyte-proliferative disorders. AB - Morphological characteristics of lymphocytes and the response to cyclosporine treatment have revealed some unique patients with pure red cell aplasia. Lymphocytes from these patients consisted mainly of non-granulated lymphocytes. All of the patients were successfully managed by cyclosporine monotherapy irrespective of prior treatment. A reduction in lymphocyte mass was not a prerequisite for the remission of pure red cell aplasia, and responses occurred within 1 month from the start of therapy. Clonal T-cell proliferation was detected in four patients, which raised the possibility of idiopathic pure red cell aplasia being associated with a clonal proliferation of T cells. An examination of the lymphocytes in patients with pure red cell aplasia could potentially be used to plan better therapeutic modalities and assess prognosis. PMID- 10460595 TI - Haemochromatosis mutations in North-East Scotland. AB - The HFE gene and its mutations C282Y and H63D cause hereditary haemochromatosis (HH). Among 54 affected individuals from North-East Scotland, 91% were homozygous for C282Y and 5.5% were compound heterozygotes for C282Y and H63D. The general population allele frequencies were high (8% and 15.7% for C282Y and H63D respectively). Although it is likely that HH is under diagnosed, these figures suggest that disease expression is variable, and many of those with the genetic predisposition HH will never develop the clinical consequences of iron overload. This has implications for diagnosis and predictive testing. PMID- 10460594 TI - High inducibility of heat shock protein 72 (hsp72) in peripheral blood mononuclear cells of aplastic anaemia patients: a reliable marker of immune mediated aplastic anaemia responsive to cyclosporine therapy. AB - To better characterize immunologic aberrations in aplastic anaemia (AA), we examined peripheral blood mononuclear cells (PBMC) of 67 patients with AA and other patients with various haematological diseases for the expression of heat shock protein 72 (hsp72), which is inducible in lymphocytes by various stressors including antigenic stimulation. When freshly obtained PBMC were examined using flow cytometry, the proportion of cells expressing hsp72 in cytoplasm was significantly higher in allogeneic marrow transplant recipients (22 +/- 15%, mean +/-standard deviation (SD)) and AA patients (17 +/- 21%) than in normal individuals (6 +/- 3%). When PBMC were tested after heat treatment, only the proportion of hsp72+ cells of AA patients (37 +/- 30%) was significantly higher than that of the normal control (17 +/- 11%). Dual fluorescence analysis of the PBMC revealed that the majority of hsp72+ cells was CD3+. For 28 untreated AA patients, the proportion of hsp72+ cells in those who later responded to cyclosporine (CyA) (62 +/- 24%) was higher than that in non-responders (19 +/- 13%). Immunoblotting analysis revealed predominant expression of hsp72 in T cells. These findings indicate that high inducibility of hsp72 in PBMC by heat treatment is an immunologic aberration characteristic of CyA-responsive AA and that this simple test may be useful for identifying a subset of AA patients responsive to immunosuppressive therapy. PMID- 10460596 TI - Coil embolization of cerebral aneurysms in patients with sickling disorders. AB - In patients with sickle cell disease cerebral aneurysm formation is thought to be a complication of recurrent red cell sickling, and multiple aneurysms have been reported in these patients. Management of patients with suspected cerebral aneurysm has traditionally involved cerebral vessel angiography followed by craniotomy and aneurysmal clipping. In patients without sickle cell disease, non operative intervention in the form of endovascular coil embolization is increasingly being used to ablate aneurysms, but has not thus far been reported in patients with sickle cell disease. We report two patients with sickling disorders who have undergone coil embolization of cerebral aneurysms with good functional and radiological outcomes. These patients illustrate that endovascular coiling is useful in the treatment of cerebral aneurysms associated with sickling disorders, although, as with surgical intervention, preparation with exchange transfusion is appropriate. PMID- 10460597 TI - Circulating platelet-neutrophil complexes represent a subpopulation of activated neutrophils primed for adhesion, phagocytosis and intracellular killing. AB - Platelets play a prominent role in linking the processes of inflammation, haemostasis and thrombosis. Recent studies have shown that platelets form heterotypic aggregates with leucocytes via platelet CD62P and leucocyte beta2 integrins. These interactions have been observed in vitro in blood taken from healthy volunteers and in clinical conditions in which thrombosis and inflammation are prominent. This study investigated the properties of platelet neutrophil complexes (PNCs) in anticoagulated whole blood. At rest, neutrophils in PNCs exhibit a significantly more activated adhesion molecule profile than free neutrophils with increased CD11b expression and activation (increased binding of the CD11b/CD18 'activation reporter' monoclonal antibody 24) and decreased CD62L expression. In addition, neutrophils in PNCs phagocytosed significantly more Neisseria meningitidis and produced more toxic oxygen metabolites than free neutrophils. Stimulation with the platelet agonist adenosine diphosphate (ADP) led to further increases in CD11b expression and activation, loss of CD62L as well as increased phagocytosis and toxic oxygen metabolite production throughout the whole neutrophil population. When these experiments were repeated with the CD62P blocking antibody G1 the effects were inhibited to a variable extent, dependent upon the parameter under investigation. These results indicate that both soluble and contact-dependent factors contribute to platelet-mediated neutrophil activation. Platelet neutrophil complexes represent a large subpopulation of neutrophils with a more activated adhesion molecule profile, and a greater capacity for phagocytosis and toxic oxygen metabolite production. This study provides further support for a role for PNCs in both health and disease. PMID- 10460598 TI - The CD69 early activation molecule is overexpressed in human bone marrow mast cells from adults with indolent systemic mast cell disease. AB - We have analysed the quantitative expression of surface CD69 antigen on human mast cells (MC), from both normal and pathological bone marrow (BM) samples, using flow cytometry. Our major aim was to analyse whether CD69 is constitutively expressed by normal BMMC and to explore the possible differences between CD69 expression by BMMC from normal controls and patients suffering from different pathological conditions. The constitutive expression of surface CD69 was clearly demonstrated in BMMC; however, systemic mast cell disease (SMCD) patients showed significantly higher levels of surface CD69 expression than healthy controls (P < 0.001), chronic lymphocytic leukaemia (P = 0.001), monoclonal gammopathy of unknown significance (P < 0.001), multiple myeloma (P < 0.001) patients, and myelodysplastic syndromes (P = 0.002). Furthermore, almost no overlap between the levels of CD69 expression on BMMC was observed between SMCD cases and the remaining groups of individuals except for the paediatric mastocytosis group (P > 0.05). From the other groups of patients, monoclonal gammopathy of unknown significance (P = 0.04), myelodysplastic syndromes (P = 0.03) and paediatric mastocytosis (P = 0.003) cases showed a significantly higher expression of surface CD69 as compared to normal subjects. In summary, our findings show that the CD69 antigen is overexpressed in SMCD patients. PMID- 10460599 TI - Induction of apoptosis and caspase activation in cells obtained from familial haemophagocytic lymphohistiocytosis patients. AB - Familial haemophagocytic lymphohistiocytosis (FHL) is a rare and uniformly fatal disorder of early childhood characterized by fever, hepatosplenomegaly, cytopenia and widespread infiltration of vital organs by activated lymphocytes and macrophages. In order to test whether the massive accumulation of immune cells in these patients is associated with a perturbation of apoptosis, lymphocytes were isolated from eight patients and subjected to the chemotherapeutic agent etoposide or agonistic anti-Fas monoclonal antibodies in vitro. These stimuli elicited a normal apoptotic response in FHL patient cells when compared to healthy controls, as determined by phosphatidylserine exposure, DNA fragmentation, in vitro cleavage of the caspase-3-like substrate aspartate glutamate-valine-aspartate-7-amino-4-methyl-coumarin (DEVD-AMC) and proteolysis of the anti-apoptotic protein Bcl-2. In addition, the degree of constitutive and Fas-triggered apoptosis in freshly isolated neutrophils was monitored in three children, with similar results. These studies indicate that immune cells derived from FHL patients are not inherently resistant to apoptosis induction. Specifically, etoposide-induced and Fas-triggered activation of intracellular caspases appears to remain intact in these individuals. However, the degree of spontaneous activation of caspase-3-like enzymes in activated lymphocytes was attenuated in three of the four patients tested prior to initiation of therapy, suggesting a possible biological deficiency in these individuals. PMID- 10460600 TI - Thrombomodulin with the Asp468Tyr mutation is expressed on the cell surface with normal cofactor activity for protein C activation. AB - Thrombomodulin (TM) is an endothelial cell glycoprotein that acts as an anticoagulant. Mutation in the TM gene is a potential risk factor for thrombosis. The first TM mutation identified was a heterozygous substitution of T for G at nucleotide position 1456, which predicted Asp468 with Tyr in a Ser/Thr-rich domain. To evaluate the reported TM gene mutation as a possible cause of thrombosis, we transiently tranfected a vector for TM gene carrying the mutation to mammalian COS7 cells. TM antigen levels in lysates of cells transfected with variant TM were comparable to those in preparations of normal TM. The TM cofactor activity for protein C (PC) activation on the variant TM-expressing cells was similar to that of the control. The Michaelis constant Km and Vmax. of variant TM for PC activation were shown to be similar compared to those of normal TM. The affinity of each TM for thrombin in PC activation was also similar. We obtained several stable cell lines expressing normal and variant TM. Lysate of the cell lines with normal and variant TM genes had a similar expression level of TM antigen. Pulse-chase analysis showed that normal and variant TM were glycosylated and resistant to endoglycosidase H, indicating that the variant TM was expressed on the cell surface in a mature form. Variant TM protein is apparently expressed on the cell surface with normal cofactor activity for PC activation. It is unlikely that the TM variant directly causes thrombosis by mechanism of reduced expression or impaired cofactor activity for PC activation, which comprises a major anticoagulant activity of TM. PMID- 10460601 TI - Reproductive choices of haemophilia carriers. AB - The actual reproductive choices made by slightly over a quarter of all the carriers of severe or moderate haemophilia in Sweden were investigated and compared with those of a randomly selected age-matched group of women who were not carriers of haemophilia. In general, the 105 carriers had the same number of children as other women of similar age. However, carriers who did not choose prenatal diagnosis (PD) often abstained from further pregnancies after the birth of a haemophilic child, and they had significantly fewer children than the remainder of the carriers, as well as fewer children than women in the control group. Logistic regression analysis showed choice of PD to be correlated to a positive attitude towards abortion following PD and a family history of haemophilia. Carriers who have experienced the complications of haemophilia or its treatment appear to be more in favour of PD than women whose haemophilic children have received modern treatment without complications. PMID- 10460602 TI - A novel ELISA-based primer extension assay for the detection of the factor V Leiden mutation. AB - We describe an enzyme-linked immunosorbent assay (ELISA) based primer extension method for the detection of the factor V Leiden (FVL) mutation. The wild-type nucleotide at position 1691 or the mutant nucleotide at the complementary position on the antisense strand were detected by the incorporation of biotinylated complementary bases onto fluorescein isothiocyanate (FITC) labelled mini-sequence primers with specificity for the sense and antisense gene segments downstream from the bases adjacent to position 1691. The reactions took place in pairs of tubes containing the complementary bases to either the wild-type or mutant nucleotide respectively. Primer extension products from each reaction tube pair which have incorporated biotinylated bases were then captured in streptavidin-coated microtitre plate wells and detected colourimetrically using an ELISA procedure. 200 patient samples were tested to validate the assay and there was complete genotypic agreement between the ELISA method and restriction site analysis using Mnl I (137 wild type, 55 FVL heterozygotes and eight homozygotes). The method utilizes non-radioactive reagents and does not require electrophoretic techniques. It is therefore a safe, simple and rapid assay which lends itself to automation. PMID- 10460603 TI - Prevalence of TT virus in plasma pools and blood products. AB - A high prevalence of TT virus (TTV), a novel virus recently identified in the serum of a patient with post-transfusion hepatitis of unknown aetiology, has been reported in blood donors worldwide. We investigated the presence of TTV DNA in several lots of blood products and in the corresponding plasma pools. In the process, we determined, from three sets of primers, the one which was most efficient in detecting the viral nucleic acid. This set amplifies the region closest to the 3'-end of the TTV genome which was proved, by sequence analysis, to be more conserved than the other two regions. Whereas all 10 intravenous immunoglobulin and 21 albumin batches were TTV negative, 4/5 factor VIII concentrates and 4/10 intramuscular immunoglobulin batches were TTV positive. A high prevalence of TTV DNA (70%) was found in the plasma pools that were collected from four different countries. These results confirm the worldwide distribution of this virus and show that TTV is removed with a varying efficiency during the manufacture of blood products. PMID- 10460604 TI - Treatment-related deaths during induction and first remission of acute myeloid leukaemia in children treated on the Tenth Medical Research Council acute myeloid leukaemia trial (MRC AML10). The MCR Childhood Leukaemia Working Party. AB - Between 1988 and 1995, 341 children with acute myeloid leukaemia (AML) were treated on the Medical Research Council Acute Myeloid Leukaemia Trial (MRC AML10). The 5-year overall survival was 57%, much improved on previous trials. However, there were 47 deaths (13. 8%), 11 of which were associated with bone marrow transplantation (BMT). The treatment-related mortality was significant at 13.8%, but decreased in the latter half of the trial from 17.8% in 1998-91 to 9. 6% in 1992-95 (P = 0.03%). The main causes of death were infection (65.9%), haemorrhage (19.1%) and cardiac failure (19.1%). Fungal infection was a significant problem, causing 23% of all infective deaths. Haemorrhage occurred early in treatment, in children with initial white cell counts >100 x 109/l (P = 0.001), and was more common in those with M4 and M5 morphology. Cardiac failure only occurred from the third course of chemotherapy onwards, with 78% (7/9) in conjunction with sepsis as a terminal event. Some deaths could be prevented by identifying those most at risk, and with prompt recognition and aggressive management of complications of treatment. Future options include the prophylactic use of antifungal agents, and the use of cardioprotectants or alternatives to conventional anthracyclines to decrease cardiac toxicity. PMID- 10460605 TI - Isolated isochromosome 17q: a distinct type of mixed myeloproliferative disorder/myelodysplastic syndrome with an aggressive clinical course. AB - A clinicopathologic study was performed on 15 patients with haematological malignancies in which isochromosome 17q [i(17q)] was the sole structural chromosome abnormality identified in bone marrow. The data indicated that an isolated i(17q) is associated with a distinct type of mixed chronic myeloproliferative/myelodysplastic disorder with an aggressive clinical course. The patients ranged in age from 37 to 83 years (median 60) with a M:F ratio of 3:1. All cases were chronic myeloid disorders with mixed proliferative and dysplastic features, making classification difficult. 11 patients tested for BCR/ABL gene fusion were normal. A low bone marrow blast count (<5%) at presentation was a typical finding. All cases had severe myeloid dysplasia which included non-segmented neutrophils and an increase in the monocyte/macrophage lineage. Fluorescence in situ hybridization (FISH) analysis of one case showed the i(17q) to involve all myeloid lineages, but not the lymphocytes. For cases with complete follow-up (n = 11) the median survival was 2.5 years (range 0.83 5.25) and 64% progressed to AML prior to death. The following features were identified which defined the haematological disorder associated with an isolated i(17q): (1) adult patient, (2) chronic myeloid disorder with clonal involvement of all myeloid lineages, (3) mixed chronic myeloproliferative/myelodysplastic features, (4) severe hyposegmentation of neutrophil nuclei, (5) prominence of the monocyte/macrophage lineage, (6) high risk for progression to AML, and (7) median survival of 2.5 years. PMID- 10460606 TI - Cross-validation of prognostic scores in myelodysplastic syndromes on 386 patients from a single institution confirms importance of cytogenetics. AB - In myelodysplastic syndromes (MDS) different prognostic risk analysis systems based on clinical and morphological data are used for predicting survival. Data on diagnostic and prognostic relevance of karyotype aberrations have prompted the development of scores including cytogenetics. The aim of this study was to assess and compare the explanatory power of different scoring systems and to assess the additional explanatory power of cytogenetics by evaluating the clinical and laboratory data of MDS patients from a single institution. Data of 386 MDS patients was available, with cytogenetic analysis at time of diagnosis in 256. Clinical/morphological scores: Bournemouth, modified Bournemouth and Dusseldorf; and scores including cytogenetics: Lausanne-Bournemouth, Lille and the International Prognostic Scoring System (IPSS), were calculated and their predictive power was compared for both overall survival and preleukaemic duration. Each of the scores had significant correlation on both endpoints. Calculating the prognostic value of different cytogenetic aberrations we found that differentiating between evidence for no aberration, single aberrations excluding chromosomes 7 and 8, aberrations on chromosomes 5, 7 or 8 and complex aberrations was important. These data were incorporated in a 'prognostic index cytogenetics' (pi score). Cytogenetic scores significantly improved the prognostic value of the best clinical/morphological score in regard to both overall survival and preleukaemic duration. In conclusion, our data further stress the importance of cytogenetics for predicting prognosis in MDS. PMID- 10460607 TI - Ineffective erythropoiesis in myelodysplastic syndromes: correlation with Fas expression but not with lack of erythropoietin receptor signal transduction. AB - Ineffective erythropoiesis in myelodysplasia is characterized by a defect in erythroid progenitor growth and by abnormal erythroid differentiation. Increased apoptosis of erythroid, granulocytic and megakaryocytic lineages is thought to account for cytopenias. Erythropoietin (Epo)-induced BFU-E and CFU-E growth was studied in 25 myelodysplastic syndrome (MDS) marrow specimens and found to be drastically diminished. To investigate the functionality of Epo-R in MDS marrow, we focused on Epo-induced STAT5 activation. Epo was able to stimulate STAT5 DNA binding activity in all normal and 12/24 MDS marrows tested, with no correlation between the level of STAT5 activation and the development of erythroid colonies in response to Epo. In contrast, impaired proliferation of erythroid progenitors was related to an increased expression of the transmembrane mediator of apoptotic cell death Fas/CD95 on the glycophorin A+ subpopulation. Therefore we conclude that the stimulation of pro-apoptotic signals rather than the defect of anti apoptotic pathways resulting from Epo-stimulated Jak2-STAT5 pathway, predominantly accounts for ineffective erythropoiesis in myelodysplasia. PMID- 10460608 TI - Response to cladribine in previously treated patients with chronic lymphocytic leukaemia identified by ex vivo assessment of drug sensitivity by DiSC assay. AB - The ability to identify non-responders to cytotoxic chemotherapy has significant clinical and economic benefits. Differential staining cytotoxicity (DiSC) assays were performed in 34 previously treated patients with chronic lymphocytic leukaemia prior to treatment with cladribine. Of the 28 identified as ex vivo sensitive, 26 achieved a complete (CR) or partial response (PR) (median length of response 1. 5 years, median survival 3.37 years) and two had a >70% fall in lymphocytes: six identified as ex vivo resistant failed to respond. The DiSC assay can accurately identify a subgroup of patients resistant to cladribine. PMID- 10460609 TI - Cyclin D2 promoter disrupted by t(12;22)(p13;q11.2) during transformation of chronic lymphocytic leukaemia to non-Hodgkin's lymphoma. AB - In a unique case of chronic lymphocytic leukaemia (CLL) we performed a longitudinal cytogenetic and molecular genetic study of tumour cells from diagnosis through progression and transformation to non-Hodgkin's lymphoma (NHL) and lymphomatous meningitis. CLL cells at diagnosis had trisomy 12 and a t(14;19)(q32;q13.3). At relapse, the leukaemic cells had a subclone carrying a t(12;22)(p13;q11.2) in addition to the initial changes. We cloned reciprocal translocation junctions at the 22q11.2- chromosome and the 12p13+ chromosome and the corresponding germline DNA fragments. Restriction map analysis and nucleotide sequence analysis of the cloned DNA fragment from the 22q11.2- chromosome mapped the translocation break within the immunoglobulin (Ig)-lambda-C complex at the nt3889; nts 3890, 3891 were lost from the translocation site. A probe from the 3' end of the clone derived from the 22q11.2- chromosome showed single copy hybridization which was different from the Ig-lambda probe. Nucleotide sequence analysis of the exact junction region and the corresponding germline DNA showed that the translocation at 12p13 occurred in the negative regulatory region of the cyclin D2 gene at the nt -1602, and a pentamer consisting of nts -1603 to -1599 was lost at the break site. We sequenced another 227 bp upstream of the known 5' end of the promoter and did not find any open reading frame. From these results we hypothesize that, in this patient, the t(12;22) disrupted the negative regulator in the promoter of cyclin D2 which in turn might have deregulated cyclin D2. PMID- 10460610 TI - Multiplex PCR reaction for the detection and identification of immunoglobulin kappa deleting element rearrangements in B-lineage leukaemias. AB - Immunoglobulin kappa (Igkappa) gene recombinations can be used - similarly to IgH rearrangements - as clonal markers in B-lineage leukaemias. Based on the extensive junctional diversity, these rearrangements represent valuable targets for the analysis of minimal residual disease (MRD). In order to provide a simple method for the rapid detection of leukaemia clone-specific kappa deleting element (Kde) mediated rearrangements, we developed a multiplex PCR reaction that is able to amplify the five most frequent rearrangements in one tube. Position of the amplimers were chosen to enable identification of the involved segments according to the size of the PCR product. This method was tested on 101 B-lineage leukaemias (71 childhood B-cell precursor acute lymphoblastic leukaemias (BCP ALL) and 30 chronic lymphocytic leukaemias (CLL)). 39 and 22 Kde rearrangements could be readily detected in 30 (44%) BCP ALL and 22 (56%) CLL, respectively. 36% of the Kde rearrangements in BCP ALL and 45% in CLL were intron recombination signal sequence (RSS)-Kde rearrangements. The other Kde rearrangements involved the Vkappa families: VkappaI in 36% and 50%, VkappaII in 32% and 16.7%, VkappaIII in 24% and 25%, and VkappaIV in 8% and 8.3% in BCP ALL and CLL, respectively. The sensitivity of the multiplex system was 10-2-10-3. We compared this multiplex PCR assay with multiple single PCR reactions using different sets of primer combinations. Thereby the number and types of rearrangements were confirmed in all cases. Clonality of rearrangements was proven by sequence analysis. Our data show that by this method clonal Kde rearrangements were rapidly detected and precisely identified. PMID- 10460611 TI - Aberrant expression and reverse signalling of CD70 on malignant B cells. AB - In normal lymphoid tissues the tumour necrosis factor-receptor family member CD27 and its ligand CD70 have a restricted expression pattern. Previously, we reported that expression of CD27 is deregulated in B-cell leukaemias and lymphomas. Here we show that, although infrequently expressed by normal human B cells in vivo, CD70 is found on 50% of B-CLLs, 33% of follicle centre lymphomas, 71% of large B cell lymphomas, and 25% of mantle cell lymphomas. Interestingly, in the majority of leukaemias and lymphomas examined, CD70 was found to have a capped appearance, a feature that coincided with co-expression of CD27. Functional analysis showed that a subset of B-CLLs could proliferate vigorously in response to CD70 mAb but not to CD27 mAb. This response was synergistically enhanced by ligation of CD40 but inhibited by the presence of IL-4. Additional experiments indicated that the proliferative response was due to an agonistic signal delivered via CD70, rather than blocking of negative signalling by CD27. Thus, next to its role as ligand, in a subset of malignant B cells CD70 can operate as receptor and as such might contribute to progression of these B-cell malignancies. PMID- 10460612 TI - Angiogenic growth factors and endostatin in non-Hodgkin's lymphoma. AB - A number of clinical studies have demonstrated the prognostic significance of angiogenesis and angiogenic growth factors in solid tumours; however, very little is known about the relevance of these parameters in haematological malignancies. We evaluated circulating levels of angiogenic growth factors and endostatin in 36 non-Hodgkin's lymphoma (NHL) patients. Baseline vascular endothelial growth factor (VEGF) levels of patients in complete remission (CR) after a median follow up of 21 months were significantly lower than those of patients with progressive disease (P = 0.016). Event-free survival (EFS) rate was significantly higher in patients who had baseline VEGF and basic-fibroblast growth factor (b.FGF) levels below the median values of 147 and 19.5 pg/ml (P = 0.018 and 0.039 by log-rank test, respectively). Conversely, the levels of endostatin, angiogenin and leptin were not different in CR patients compared to relapsed patients and did not correlate with EFS. Our data suggest that b-FGF and, particularly, VEGF might be considered prognostic factors in NHL staging and management. PMID- 10460613 TI - GBV-C/HGV and HCV infection in mixed cryoglobulinaemia. AB - Recently, a new, suspected hepatotropic virus has been identified. Named GBV C/HGV, this virus shares with the hepatitis C virus (HCV) routes of transmission and molecular organization. Indeed, a proportion of HCV-infected patients (10 25%) are also carriers of GBV-C/HGV. Since mixed cryoglobulinaemia (MC) is closely associated with HCV infection, the aim of this study was to determine the prevalence of GBV-C/HGV infection in MC patients, and to investigate whether the double infection influenced the clinical and/or laboratory aspects of the disease. 52 patients affected by MC were studied. 100 patients affected by HCV positive chronic liver disease (CLD) without MC were used as control group. To determine the prevalence of GBV-C/HGV infection in general population, 150 blood donors were studied, as well as 80 patients affected by non-A-E CLD. Among the MC patients, only five (9.6%) were positive for both HCV and GBV-C/HGV infection. No difference was found between patients with and without double infection as regards main clinical and laboratory aspects. Among HCV-positive CLD cases, 27 were positive for double infection. Among blood donors, the prevalence of GBV C/HGV infection was 8.0%, whereas in cases with cryptogenetic CLD the prevalence was 5.0%. In conclusion, these data show that GBV-C/HGV infection does not play any role in the pathogenesis of MC. PMID- 10460614 TI - Long-term follow-up of patients with hairy cell leukaemia after treatment with pentostatin or cladribine. AB - We report the long-term follow-up results on two groups of patients with hairy cell leukaemia (HCL) treated with either pentostatin (deoxycoformycin) or cladribine (2-chlorodeoxyadenosine). 165 HCL patients received treatment with pentostatin (between 1986 and 1994), and 45 were treated with cladribine (between 1992 and 1997). Age and sex characteristics were similar in the two groups. 38 patients in the pentostatin group and 12 in the cladribine group were previously untreated. 22 patients in the cladribine group had received prior treatment with pentostatin; four were resistant, 17 had relapsed following partial (four) or complete (13) responses, and one was not evaluable for response. The response rates were the same in the two groups: 82% complete response (CR), 15% partial response (PR) for pentostatin and 84% CR, 16% PR for cladribine. Relapse rates were 24% for pentostatin and 29% for cladribine after median follow-up of 71 and 45 months respectively. At 45 months, however, the relapse rate for pentostatin was only 9.7%. We found a statistically significant difference in the disease free interval (DFI) between the two groups suggesting that patients may relapse more quickly after cladribine. The majority of relapsed patients achieved second remissions following further therapy with either pentostatin or cladribine, with no evidence of cross resistance between the two agents. The 5-year survival for all patients was 97% and treatment- related toxicity was low. We conclude that both pentostatin and cladribine induce durable remissions in the majority of HCL patients. Longer follow-up is required to establish whether some patients are cured as there is no plateau in DFI, and which of these two agents may be the treatment of choice. PMID- 10460615 TI - Complete remission in acute myeloid leukaemia with t(8;21) following treatment with G-CSF: flow cytometric analysis of in vivo and in vitro effects on cell maturation. AB - A 75-year-old patient diagnosed as having acute myeloid leukaemia with t(8;21) received G-CSF alone as induction therapy. Complete remission was achieved following 2 weeks of treatment. Flow cytometric analysis, performed by CD45 technique modified by the introduction of preliminary gating with LDS-751, confirmed the disappearance of blast cells along with myeloid maturation. Finally, in vitro studies demonstrated that G-CSF, as compared to other differentiation inducers, was able to induce a striking effect toward neutrophilic differentiation. PMID- 10460616 TI - alpha4beta1 integrin-mediated adhesion of CD34+ cells from patients with chronic myeloid leukaemia: influence of IL-3. AB - Interactions between integrins on haemopoietic progenitor cells and their stromal ligands have an important role in the control of haemopoiesis. Growth factors can modulate these interactions (so-called 'inside-out' signalling) resulting in changes in ligand binding activity. We have studied alpha4beta1 integrin-mediated adhesion to the H120 fragment of fibronectin (which contains the strongest alpha4beta1 binding site) in CD34+ cells from patients with chronic myeloid leukaemia (CML) and have determined the effect of IL-3 on the level of adhesion. Compared to normal CD34+ cells isolated from cord blood and peripheral blood progenitor harvests (mean of 61.4 +/- 14.9% of cells attached) the CML CD34+ cells showed reduced levels of adhesion (mean of 41.9 +/- 14.7%, P < 0.05). The effect of 10 ng/ml of IL-3 resulting in reduced adhesion of normal CD34+ cells at 30 min was absent in 6/7 patients with CML. Abnormalities of adhesion to fibronectin may thus be related to IL-3 pathways affected by BCR-ABL. These findings will have implications for understanding the dysregulation of growth and adhesion in CML. PMID- 10460617 TI - Detection of a Hodgkin/Reed-Sternberg cell specific immunoglobulin gene rearrangement in the serum DNA of a patient with Hodgkin's disease. AB - We analysed multiple serum samples from a patient with mixed cellularity Hodgkin's disease for the Hodgkin/Reed-Sternberg cell clone-specific rearranged Ig gene sequence. The clone-specific sequence could be detected in DNA extracted from a serum sample obtained during clinical relapse but not in serum samples obtained during or after treatment following relapse. PMID- 10460618 TI - CD130 rather than CD126 expression is associated with disease activity in multiple myeloma. AB - We analysed the expression of both components of IL-6R, CD126 the ligand binding protein and CD130 the signal transducing protein, on plasma cells from MGUS and multiple myeloma (MM) cases using flow cytometry. CD126 was detectable in 50% of either MGUS or MM patients without any change of expression during disease progression. In contrast, CD130 expression was up-regulated during tumoural expansion (43% of MM patients at diagnosis versus 88% at relapse). Finally, combining CD126 and CD130 expression we found a significant increase of the percentage of CD126+ CD130+ patients at relapse, underlying the crucial role of IL-6 response in the late stage of MM. PMID- 10460619 TI - Early diagnosis of central nervous system aspergillosis using polymerase chain reaction, latex agglutination test, and enzyme-linked immunosorbent assay. AB - To investigate the usefulness of examination of cerebrospinal fluids (CSF) in the diagnosis of central nervous system (CNS) aspergillosis, we examined five patients with either brain abscesses or cerebral infarctions and 11 control patients. CSF samples were subjected to enzyme-linked immunosorbent assay (EIA), latex agglutination test (LA) and polymerase chain reaction (PCR). Cultures of CSF samples were negative in all the patients, but PCR, EIA and LA were positive in five, four and four patients with CNS aspergillosis, respectively. None of these tests were positive in the control patients. CSF examination may be beneficial in the diagnosis of CNS aspergillosis. PMID- 10460620 TI - Use of the haemopoietic progenitor cell count of the Sysmex SE-9500 to refine apheresis timing of peripheral blood stem cells. AB - The Sysmex SE-9500 automated cell counter provides an estimate of immature cells referred to as 'haemopoietic progenitor cells' (HPC). The aim of this study was to relate the HPC count to CD34+ cell levels in mobilized peripheral blood and to determine whether the HPC count was valuable in predicting apheresis yields of CD34+ cells. Studies were performed on 114 samples from 67 patients undergoing progenitor cell mobilization. HPC cells were undetectable in the steady state. On the day of apheresis the HPC and CD34 counts were weakly correlated, with the median HPC count being 2.3-fold greater than the CD34+ cell count. The HPC count did not include the CD34+ cells as immunomagnetic depletion of CD34+ cells did not significantly reduce the HPC count. CD34+ cell counts predicted for apheresis yield (r = 0.773) on that day as did the HPC count (r = 0. 623). The optimal strategy to prevent unnecessary harvesting while minimizing the risk of missing an adequate harvest, and minimizing laboratory investigations, was to screen all samples for HPC and limit CD34+ cell measurements to those with an HPC count <10 x 106/l (19/114 samples). If the CD34+ cell count was also <10 x 106/l then harvesting should not be carried out. PMID- 10460621 TI - Clonotypic CD20+ and CD19+ B cells in peripheral blood of patients with multiple myeloma post high-dose therapy and peripheral blood stem cell transplantation. AB - The number of circulating clonotypic B cells in patients with multiple myeloma (MM) after high-dose therapy (HDT) with peripheral blood stem cell transplantation (PBSCT) was investigated. Peripheral CD19+ B cells have been reported to persist throughout conventional and HDT and might resemble a source of relapse in patients with MM. We assessed the proportion of malignant cells in CD20+ and CD19+ cell fractions of 14 peripheral blood (PB) samples from 12 patients after HDT and PBSCT. Nine samples were obtained from patients in continuous remission, and five patients were in progressive disease or beginning relapse. The CD20+ fractions obtained had a mean purity of 96.8%. The percentages of tumour cells were determined using a quantitative allele-specific oligonucleotide PCR assay based on the method of limiting dilutions. In the group of patients in continuous remission the median number of tumour cells in the CD20+ cell fractions was 1.9/ml (range 0-7.2 tumour cells/ml PB) higher than in the CD20- fractions (median 0; range 0-29 tumour cells/ml PB). Higher tumour cell numbers in both fractions, particularly pronounced in the negative ones, were found in patients with progressive disease or beginning relapse (CD20+: range 3.8 585; median 32 tumour cells/ml PB; CD20-: range 25-25527; median 334 tumour cells/ml PB). Enrichment with the anti-CD19 antibody as a second pan B-cell marker revealed comparable tumour cell numbers. In conclusion, an anti-CD20 antibody treatment could be a promising approach for the eradication of malignant cells in the PB of patients in continuous remission after HDT and PBSCT with low amounts of tumour cells in the B-cell compartment and an almost complete absence of tumour cells in the CD20- fractions. PMID- 10460622 TI - A combination of megakaryocyte growth and development factor and interleukin-1 is sufficient to culture large numbers of megakaryocytic progenitors and megakaryocytes for transfusion purposes. AB - Chemotherapy-induced thrombocytopenia is a major risk factor in cancer treatment. The transfusion of autologous ex vivo expanded megakaryocytes could be a new therapy to shorten the period of thrombocytopenia. Therefore we investigated, in a liquid culture system, the effect of various cytokine combinations composed of pegylated megakaryocyte growth and development factor (PEG-rHuMGDF), interleukin 1 (IL-1), IL-3, IL-6, IL-11 and stem cell factor (SCF) on the proliferation and differentiation of CD34+ cells, in order to define the most optimal and minimum levels of cytokine combinations for megakaryocyte expansion. Besides PEG-rHuMGDF, IL-1 was found to be important for optimal megakaryocyte expansion. Depletion of either SCF, IL-6 or IL-11 did not exert a large effect, but the absence of IL-1 strongly diminished the number of megakaryocytic cells. Addition of IL-3 to the combination PEG-rHuMGDF, IL-1, IL-6, IL-11 and SCF significantly reduced the number of megakaryocyte progenitors (CD34+CD41+ cells) and the number of CFU-Meg. Furthermore, we found a strong correlation between the number of CD34+CD41+ cells and the number of CFU-Meg obtained after 8 d culture. Our study shows that optimal ex vivo expansion of megakaryocytes is achieved by the combination of PEG rHuMGDF and IL-1. The numbers of megakaryocytes and megakaryocyte progenitors (CD34+CD41+) obtained in our liquid culture system with the growth factor combination PEG-rHuMGDF and IL-1 are suitable for transfusion purposes. PMID- 10460623 TI - Prevalence of mild hyperhomocysteinaemia and association with thrombophilic genotypes (factor V Leiden and prothrombin G20210A) in Italian patients with venous thromboembolic disease. AB - Mild hyperhomocysteinaemia is an established risk factor for deep vein thrombosis (DVT); few data concerning its potential interaction with thrombophilic genotypes are available at the present time. We investigated 121 thrombosis-free individuals and 111 patients with at least one objectively confirmed episode of DVT. A thrombophilic condition (deficiency in antithrombin, protein C and S, factor V Leiden, prothrombin G20210A) was detected in 25.2% of the patients; mutant factor V or prothrombin genotypes were present in 6.6% of the controls. Hyperhomocysteinaemia was found in 14.4% of patients and 3. 3% of the controls, with a 3.7-fold increase in risk for DVT (95% CI 1.1-12.3). Adoption of different cut-off levels for definition of hyperhomocysteinaemia did not substantially change the magnitude of the risk. Carriership of both hyperhomocysteinaemia and factor V Leiden or prothrombin G20210A was detected in 2.7% of patients for each combination and in none of the controls. An approximate estimate of 30-fold increased risk in carriers of both hyperhomocysteinaemia and factor V Leiden and 50-fold increased risk in carriers of both hyperhomocysteinaemia and prothrombin G20210A was calculated, suggesting a synergistic interaction between hyperhomocysteinaemia and such thrombophilic genotypes. Yet statistical analysis is highly unstable due to the small number of individuals with combined defects. Further investigations on large series of patients are needed. PMID- 10460624 TI - Successful management of concomitant Diamond-Blackfan anaemia and aplastic anaemia with splenectomy. PMID- 10460625 TI - An atypical myelodysplastic syndrome with t(9;12)(q22;p12) and TEL gene rearrangement. PMID- 10460626 TI - Clonal selection of CD20-negative non-Hodgkin's lymphoma cells after treatment with anti-CD20 antibody rituximab. PMID- 10460627 TI - Calcitriol in myelodysplastic syndromes. PMID- 10460628 TI - rHuEpo treatment in low-risk myelodysplastic syndromes. PMID- 10460629 TI - rHuEpo TREATMENT IN LOW-RISK MYELODYSPLASTIC SYNDROMES: A REPLY PMID- 10460630 TI - Systematic reviews and meta-analyses: the value for surgery. PMID- 10460631 TI - Postoperative small bowel leak. PMID- 10460632 TI - Adjuvant medical therapy in peripheral bypass surgery. AB - BACKGROUND: A review was conducted of published clinical trials of adjuvant medical therapy in infrainguinal bypass procedures to evaluate the strength of the evidence for the use of various agents. METHODS: Trials were identified by literature search. The methods used were reviewed and the results with each agent tested were assessed taking into account the soundness of the study design. RESULTS: Thirty-three studies were identified; fewer than half had a randomized and double-blind design. Most were single-centre studies including a mixture of different surgical procedures and patients with varying degrees of lower limb ischaemia. Clinical outcomes were seldom reported. The median sample size was 61. The median follow-up duration was 12 months, but was often not standardized for all patients in a trial. Only aspirin in prosthetic grafts and ticlopidine in vein grafts have been shown in well designed, double-blind, randomized, controlled trials to reduce the likelihood of occlusion in infrainguinal bypass grafts. CONCLUSION: The majority of the trials reviewed had significant deficiencies in their design, reducing the reliance that can be placed on their results. Further studies are required to investigate adequately the effectiveness of existing medical therapies for the maintenance of infrainguinal bypass grafts. PMID- 10460633 TI - Graduated compression stockings in the prevention of venous thromboembolism. AB - BACKGROUND: Surveys still show a wide variation in routine use of deep vein thrombosis (DVT) prophylaxis despite its established place in current patient management. This article reviews the mechanism of action, efficacy and complications of stockings in preventing DVT. METHODS: Relevant publications indexed in Medline (1966-1998) and the Cochrane database were identified. Appropriate articles identified from the reference lists of the above searches were also selected and reviewed. RESULTS AND CONCLUSION: Graduated compression stockings reduce the overall cross-sectional area of the limb, increase the linear velocity of venous flow, reduce venous wall distension and improve valvular function. Fifteen randomized controlled trials of graduated compression stockings alone were reviewed. Stockings reduced the relative risk of DVT by 64 per cent in general surgical patients and 57 per cent following total hip replacement. The effect of stockings was enhanced by combination with pharmacological agents such as heparin; the combination is recommended in patients at moderate or high risk of DVT. Knee-length stockings are as effective and should replace above-knee stockings. Complications are rare and avoidable. PMID- 10460634 TI - Experimental study of liver dysfunction evaluated by direct indocyanine green clearance using near infrared spectroscopy. AB - BACKGROUND: Blood clearance of indocyanine green (ICG) is an objective test of liver function. Hepatic ICG clearance can now be measured directly using near infrared spectroscopy (NIRS). The aim of this study was to evaluate measurement of hepatic ICG clearance by NIRS in an animal model of acute hepatic dysfunction. METHODS: New Zealand white rabbits (n = 36) underwent laparotomy for liver exposure. Hepatic blood flow and microcirculation were measured along with hepatic ICG concentration by NIRS. Hepatic ICG clearance was measured in groups of six animals after reduction of the hepatic blood flow by hepatic artery occlusion and portal vein partial occlusion, lobar ischaemia and reperfusion (I/R), colchicine administration and bile duct ligation. Hepatic ICG uptake and excretion rates were calculated by a non-linear least square curve fitting method from the ICG concentration-time curve. RESULTS: There was a significant positive correlation between hepatic ICG rate of uptake and both hepatic blood flow and microcirculation (r = 0.79, P = 0.0001; r = 0.59, P = 0.005 respectively). I/R resulted in a significant reduction of both the rates of ICG uptake (mean(s.d.) 0. 85(0.59) min-1; P = 0.0002 versus control) and ICG excretion (0. 020(0.006) min-1; P = 0.02 versus control). Colchicine decreased the rate of hepatic ICG excretion (0.030(0.010) min-1; P = 0.02 versus control) as did bile duct ligation (0.002(0.001) min-1; P = 0.01 versus control). CONCLUSION: Measurement of hepatic ICG clearance by NIRS is a promising technique for assessing hepatic parenchymal dysfunction and may have application in liver surgery and transplantation. PMID- 10460635 TI - Major hepatectomy for hepatocellular carcinoma in patients with an unsatisfactory indocyanine green clearance test. AB - BACKGROUND: Liver failure is the commonest cause of postoperative death in patients with hepatocellular carcinoma (HCC). With the improvement in operative technique and perioperative care, the limit of hepatic functional reserve may be lowered. The aim of this study was to evaluate the postoperative morbidity, mortality and survival rates in patients with an indocyanine green (ICG) retention value higher than 14 per cent, after major hepatectomy for HCC. METHODS: From January 1994 to December 1997, 117 patients underwent major hepatectomy for HCC; 92 patients had preoperative ICG retention at 15 min lower than 14 per cent (median 8.3 (range 1.6-13.8) per cent), while 25 patients had ICG retention greater than 14 per cent (17.4 (range 14.3-35.3) per cent). Data were collected prospectively and analysed retrospectively. RESULTS: The two groups of patients were similar in terms of age, sex ratio, preoperative platelet count, liver biochemistry, Child-Pugh status and operative procedures performed, but the prothrombin time was significantly longer in the high ICG group. The operative blood loss (1.5 litres), the amount of blood transfused and the number of patients requiring blood transfusion were similar. The postoperative complication rate (41 versus 40 per cent), duration of hospital stay (12 versus 13 days), hospital mortality rate (1 versus 4 per cent) and median survival time (47 versus 45 months) were not significantly different. CONCLUSION: With meticulous surgical technique to decrease intraoperative blood loss and good perioperative care, selected patients with limited hepatic functional reserve can achieve a good immediate postoperative result and a survival rate similar to that of patients with good hepatic functional reserve. PMID- 10460636 TI - Management of pancreatic remnant with strategies according to the size of pancreatic duct after pancreaticoduodenectomy. PMID- 10460637 TI - Bacterial infection and extent of necrosis are determinants of organ failure in patients with acute necrotizing pancreatitis. AB - BACKGROUND: The risk factors predisposing to organ failure in patients with necrotizing pancreatitis remain unclear. The relationship between the extent of pancreatic necrosis, the presence of infection and the incidence of organ failure was analysed. METHODS: In a retrospective review, the occurrence of pulmonary insufficiency, renal insufficiency, shock, sepsis/sepsis-like syndrome (SLS) and coagulopathy was evaluated in 273 patients with necrotizing pancreatitis, and a comparison was made between patients with sterile or infected necrosis. Additionally, the relation between the incidence of organ failure and extent of pancreatic parenchymal necrosis was investigated by classifying the patients into three groups according to the amount of necrotic tissue found by contrast enhanced computed tomography (group 1, extent less than 30 per cent; group 2, 30 50 per cent; group 3, more than 50 per cent). RESULTS: Organ failure was more frequent in patients with infected necrosis than in those with sterile necrosis. Differences were found in the incidence of pulmonary insufficiency, sepsis/SLS and coagulopathy. Organ failure occurred more frequently in group 3 than in group 2 or 1 (95 versus 79 and 66 per cent; P = 0.0004). The extent of infected necrosis was not related to the incidence of organ failure (group 1, 88 per cent; group 2, 86 per cent; group 3, 96 per cent). However, there was a relation between the incidence of organ failure and the extent of sterile necrosis (group 1, 59 per cent; group 2, 74 per cent; group 3, 94 per cent; P = 0.0001). Multivariate analysis confirmed the presence of infection and the extent of necrosis as independent determinants of organ failure. CONCLUSION: The incidence of organ failure is determined by both bacterial infection and extent of necrosis. The incidence of organ failure is determined by the extent of necrotic parenchyma in patients with sterile necrosis. Infected necrosis is associated with a high incidence of organ failure irrespective of the extent of necrosis. PMID- 10460638 TI - Adjuvant regional chemotherapy after hepatic resection for colorectal metastases. AB - BACKGROUND: This study explored the possibility of achieving a better survival rate and reduced recurrence in the remaining liver in patients with colorectal hepatic metastases undergoing hepatic resection. Adjuvant postoperative regional chemotherapy was administered via the hepatic artery or the portal vein. METHODS: A retrospective study was performed on 174 patients after hepatic resection for colorectal metastases. These comprised 78 patients who had hepatic artery infusion (HAI) chemotherapy (HAI group), 30 who had portal vein infusion (PVI) chemotherapy (PVI group) and 66 who had no regional chemotherapy (resection alone group). The three groups were compared with one another in terms of complications, survival rate and patterns of recurrence. RESULTS: Severe complications did not occur at any point during adjuvant HAI or PVI chemotherapy. The 5-year disease-free survival rate of patients in the HAI, PVI and resection alone groups were 35, 13 and 9 per cent respectively, including six hospital deaths. Patients in the HAI group showed significantly improved recurrence rates in the remaining liver compared with the resection alone group (P = 0.03), and more prolonged disease-free and overall survival than those in the PVI (P = 0.01 and P = 0.02 respectively) and resection alone (P = 0.0001 and P = 0.0006 respectively) groups. CONCLUSION: This study suggests that adjuvant HAI chemotherapy after hepatic resection may have therapeutic potential for improved management of patients with colorectal metastases. PMID- 10460639 TI - Prognostic significance of anatomical resection and des-gamma-carboxy prothrombin in patients with hepatocellular carcinoma. AB - BACKGROUND: Portal venous tumour extension and intrahepatic metastasis result in a poor prognosis following hepatectomy for hepatocellular carcinoma (HCC). Anatomical resection is, in theory, preferable for eradicating these types of invasion. Des-gamma-carboxy prothrombin (DCP) has been reported to be associated with adverse pathological variables. This study investigated the significance of anatomical resection and DCP as predictive factors for postoperative recurrence of HCC. METHODS: A retrospective cohort study was carried out in 138 consecutive patients who underwent hepatectomy for HCC smaller than 5 cm using the Cox proportional hazards model. RESULTS: Eight factors were univariately related to poor prognosis (in decreasing order of hazard ratio): intrahepatic metastasis, multiple tumours, alpha-fetoprotein 32 ng/ml or more; DCP greater than 0.1 arbitrary units (AU), tumour-exposed surgical margin, vascular invasion, non anatomical resection and tumour 2.5 cm or more. Three variables (DCP, vascular invasion and tumour-exposed surgical margin) were excluded by a stepwise procedure in multivariate analysis. Although DCP was not an independent prognostic factor, a model replacing intrahepatic metastasis with DCP showed similar predictive accuracy in a receiver-operating characteristic curve. CONCLUSION: Anatomical resection appeared to have a beneficial effect on recurrence-free survival after hepatectomy for HCC. DCP measurement was effective in predicting HCC recurrence and had the advantage that it can be assessed before operation. PMID- 10460640 TI - Biphasic role for nitric oxide in experimental renal warm ischaemia-reperfusion injury. AB - BACKGROUND: Whilst nitric oxide has a clearly defined role in renal haemostasis, debate continues over its pathophysiology. This study investigated the function of nitric oxide in a model of renal warm ischaemia-reperfusion injury. METHODS: Rats underwent bilateral renal warm ischaemia (45 min) after pretreatment with nitric oxide donors, nitric oxide synthase (NOS) inhibitors or saline (control). Following reperfusion (20 min) a unilateral nephrectomy was performed to measure renal nitric oxide (as nitroxides) and oxidative DNA and protein damage. Renal function was measured on days 2 and 7 before terminal nephrectomy for analysis and morphology. RESULTS: The increase in renal nitric oxide level seen early in reperfusion (20 min) (P < 0.01) was prevented by inhibition of constitutive (cNOS) but not inducible (iNOS) NOS. The increase in oxidative damage (P < 0.01) was exacerbated by nitric oxide donors (P < 0.01) but ameliorated by NOS inhibition (P < 0.01). Control nitric oxide remained increased through to day 7 (P < 0.01) but was reduced by nitric oxide donors and cNOS inhibitors (P < 0.05). Oxidative damage returned towards normal in the control group, whereas both DNA and protein damage persisted following NOS inhibition (P < 0.01). CONCLUSION: Inhibition of the postischaemic increase in the level of nitric oxide was associated with an early decrease in, but eventual exacerbation of, oxidative damage. This suggests the prolonged increase in renal nitric oxide concentration was cytoprotective overall. PMID- 10460641 TI - Factors affecting late survival after elective abdominal aortic aneurysm repair. AB - BACKGROUND: The late prognosis after repair of an abdominal aortic aneurysm (AAA) may be affected by atherosclerosis, which involves various organs including the aorta. METHODS: The influence of organ dysfunction and atherosclerosis on the long-term survival of 338 consecutive patients undergoing successful elective AAA repair between 1980 and 1997 was analysed using Cox hazards model. Survival rate was compared with that of 349 patients undergoing successful revascularization for aortoiliac occlusive disease (AIOD) during the same interval. RESULTS: Renal dysfunction and a previous history of cerebrovascular events were important variables predicting late death (risk ratio 1.980 and 1.903 respectively), while a history of cardiac disease predicted only cardiac-related death. The survival rate in patients with normal renal function was significantly better than that in those with renal dysfunction (P = 0.0371). Similarly, the survival rate was significantly better in patients without a history of previous cerebrovascular events (P = 0.0414). The survival rate after AAA repair was nearly identical to that of age- and sex-matched patients with grade II or III symptoms who had surgery for AIOD. CONCLUSION: Advanced atherosclerosis with underlying organ dysfunction is a true determinant of long-term survival following elective repair of AAA. PMID- 10460642 TI - Marimastat inhibits elastin degradation and matrix metalloproteinase 2 activity in a model of aneurysm disease. AB - BACKGROUND: Abdominal aortic aneurysms are characterized by degradation of the extracellular matrix, with a reduction in the elastin concentration of the arterial media. These changes have been linked to increased levels of endogenous metalloproteinases (MMPs) within the aorta, particularly MMP-2 and MMP-9. This provides a potential therapeutic target for pharmacological agents aimed at reducing the growth rate of small aneurysms. In this study, the ability of marimastat (an MMP inhibitor) to reduce matrix degradation was assessed in a previously described model of aneurysm disease. METHODS: Porcine aortic segments (n = 12) were preincubated in exogenous pancreatic elastase for 24 h before culture in standard conditions for 13 days with marimastat 10(-5), 10(-6) and 10( 7) mol/l. Control segments were cultured both without marimastat and without elastase. At the termination of culture, MMPs were extracted from the tissue and quantified by substrate gel enzymography. The volume fractions of elastin and collagen were determined by stereological analysis of sections stained with Miller's elastin and van Gieson's stain. RESULTS: Stereological analysis demonstrated preservation of elastin in aorta treated with marimastat at 10(-6) and 10(-5) mol/l; this was significant at the latter concentration (P = 0.007). This was accompanied by a significant reduction in active MMP-2 activity in the samples treated with marimastat 10(-5) mol/l (P < 0.01). CONCLUSION: Marimastat significantly inhibited elastin degradation and active MMP-2 production within aortic organ cultures. PMID- 10460643 TI - Mesenteric vein thrombosis due to factor V Leiden gene mutation. AB - BACKGROUND: Mesenteric venous thrombosis is a rare cause of acute abdominal pain that may be the result of coagulation abnormalities. METHODS: Four consecutive patients with mesenteric venous thrombosis underwent haematological evaluation. RESULTS: All four had activated protein C resistance resulting from a single mutation in the gene coding for coagulation factor V. Three had surgery; in one patient the diagnosis was made by ultrasonography. One of the patients who had surgery died but the other three survived and were treated with long-term anticoagulation. CONCLUSION: Activated protein C resistance may be an important pathogenetic factor in primary mesen-teric vein thrombosis. PMID- 10460644 TI - Outcome after surgical resection for high-grade anal intraepithelial neoplasia (Bowen's disease). AB - BACKGROUND: High-grade anal intraepithelial neoplasia (Bowen's disease) may predispose to anal carcinoma. Treatment options include surgical resection but effectiveness remains uncertain. This paper reports long-term follow-up of patients with high-grade anal intraepithelial neoplasia treated by surgical resection. METHODS: Between 1989 and 1996, 46 patients were identified with high grade anal intraepithelial neoplasia. Thirty-four underwent local excision of all macroscopically abnormal disease and the resulting defect was left open, closed primarily or skin grafted. Regular follow-up subsequently included anoscopy and biopsy of any suspicious lesions. RESULTS: Median follow-up was 41 (range 12-104) months. Total excision was difficult; 19 patients had histological evidence of incomplete excision at the time of initial resection. Some 12 of 19 had histo logically proven recurrent high-grade intraepithelial neoplasia within 1 year. Even with microscopically complete excision two of 15 patients subsequently developed recurrent high-grade intraepithelial neoplasia at 6 and 32 months after operation. No patient developed carcinoma but five had complica-tions of anal stenosis or faecal incontinence. CONCLUSION: Although no definite recommendations can be made for the treatment of high-grade anal intraepithelial neoplasia, these results illustrate some potential drawbacks of surgical excision with a high potential for incomplete excision and persistent disease, even after complete excision in some patients, and a high morbidity rate. PMID- 10460645 TI - Association between alcoholism and diverticulitis. PMID- 10460646 TI - Smoking is a predictive factor for outcome after colectomy and ileorectal anastomosis in patients with Crohn's colitis. PMID- 10460647 TI - Unstimulated graciloplasty in traumatic faecal incontinence. PMID- 10460648 TI - Oral contrast with computed tomography in the evaluation of blunt abdominal trauma in children. AB - BACKGROUND: The use of oral contrast in evaluating children by computed tomography (CT) following blunt trauma is controversial. The aim of this study was to evaluate retrospectively the use of oral contrast with abdominal CT in children with suspected abdominal injury. METHODS: The medical records of 101 children who underwent CT for abdominal trauma between 1993 and 1997 were reviewed for data pertaining to the mechanism of injury, clinical findings and management. Scans were reviewed by a paediatric radiologist and criteria of intestinal injury on CT described by Cox and Kuhn were used: (1) extraluminal air or contrast material, (2) focal area of thickening of bowel wall and mesentery, and (3) free intraperitoneal fluid in the absence of solid organ injury. RESULTS: CT was performed within a median time of 2.4 (range 1-48) h after the injury. On 37 (62 per cent) of 60 scans in children who had oral contrast, the duodenum was not opacified after a mean delay of 30 min. Intestinal injury was suspected on CT in four children. In two children with CT evidence of intestinal injury (with/without oral contrast) rupture of the duodenojejunal flexure (n = 1) or ileal perforation (n = 1) was found at laparotomy. Two children had a false positive scan, leading to negative laparotomy; one scan with oral contrast incorrectly suggested a duodenal leak and in another child CT without oral contrast showed thickening of bowel wall with free intraperitoneal fluid but no specific intestinal injury was identified at laparotomy. One patient had two negative CT scans (with and without oral contrast) and underwent laparotomy for clinical suspicion of bowel injury; rupture of the splenic flexure of the colon was found at laparotomy. CONCLUSION: CT is not reliable for diagnosing intestinal injuries and this is not improved by use of oral contrast. Omission of oral contrast was not associated with delay in the diagnosis of intestinal injury. Since intestinal injuries are uncommon in children, a prospective multicentre study would determine more precisely the role of the routine use of oral contrast. PMID- 10460649 TI - Consultant surgeons' opinion of the skills required of basic surgical trainees. AB - BACKGROUND: Accurate and appropriate assessment of surgical trainees requires clear determination of the skills needed for surgical competence. This study was designed to identify those skills, rank them in order of importance and translate them into behavioural terms. METHODS: A Delphi technique, using anonymous postal questionnaires, was used. All consultant surgeons in South-East Scotland were asked to identify the skills they expected of surgical trainees. Skills identified were then returned to all consultants for weighting. Differences among specialties in the importance of each item were identified using analysis of variance. RESULTS: The qualities identified fell into five domains: technical skills, clinical skills, interaction with patients and relatives, teamwork, and application of knowledge. Consultants from all specialties gave high weightings to the generic domains of clinical skills, teamwork, and interaction with patients and relatives. CONCLUSION: This study has identified the skills considered necessary by consultant surgeons in Scotland for a successful surgical career. Contrary to expectation, consultant surgeons value many generic skills more highly than technical skills, indicating that they value well rounded doctors, not just those with technical ability. The characteristics identified are being used to develop an assessment tool for use on basic surgical trainees. PMID- 10460650 TI - Role of laparoscopic ultrasonography in the management of patients with oesophagogastric cancer. AB - BACKGROUND: Laparoscopy and laparoscopic ultrasonography (lapUS) have been shown to improve the staging of patients with oesophagogastric cancer but there remains doubt as to whether most benefit follows laparoscopy alone and how much is contributed by the addition of lapUS. METHODS: The role of lapUS in surgical decision making was evaluated prospectively in a consecutive series of patients with oesophagogastric cancer following conventional radiological assessment. The results of the lapUS findings over and above the laparoscopic findings were documented in order to identify the additional benefit of lapUS. RESULTS: After initial conventional assessment 41 patients were considered unsuitable for surgery and treated by palliation, with a further 25 patients proceeding to surgery without laparoscopy. Of the 93 patients who underwent laparoscopy, 18 were shown to have irresectable disease and avoided further surgery; a further seven avoided inappropriate surgery by the addition of lapUS. The open-close laparotomy rate was reduced from five of 25 in patients who did not undergo laparoscopy to nine (12 per cent) of 75 by the introduction of laparoscopy and to two (3 per cent) of 68 with the addition of lapUS. CONCLUSION: Laparoscopy alone prevented unnecessary surgery in 18 (19 per cent) of 93 patients with oesophagogastric cancer and the addition of lapUS identified a further seven patients (8 per cent) in whom unnecessary surgery was avoided. PMID- 10460652 TI - Electron microscopy for tumour diagnosis: is it redundant? AB - The histopathological diagnosis of tumours has been transformed by immunohistochemistry. Used with experience and judgement, a panel of antibodies or antisera, combined when necessary with antigen retrieval, will enable the accurate typing of most problematic tumours. This has led many histopathologists to question whether the electron microscope has any residual utility for tumour diagnosis; the machines are large, costly to purchase and maintain, and will accept only minute samples of tissue. The following articles by Mierau and by Eyden, both strong advocates, comment on the current and future role of electron microscopy in tumour diagnosis. PMID- 10460651 TI - Results of pneumatic dilatation in patients with dysphagia after antireflux surgery. AB - BACKGROUND: The mechanisms and treatment of persistent dysphagia after antireflux surgery are not well established. The results of pneumatic dilatation were evaluated in a retrospective study. METHODS: Sixteen patients were reviewed. All had severe and persistent postoperative dysphagia evaluated by oesophageal manometry before pneumatic dilatation. RESULTS: Seven patients had one dilatation and nine had two dilatations. There was no complication and no relapse of reflux symptoms. Results were satisfactory in nine patients (mean(s.e.m.) follow-up 19.2(6.9) months) and poor in seven, who required revisional surgery. Age, time since operation and weight loss were not related to the outcome. Thirteen patients had abnormal manometry with an 'achalasia-like' motor pattern in four. Lower oesophageal sphincter (LOS) pressure, LOS relaxation and oesophageal contraction amplitude were similar in the two groups. The only difference was in the percentage of normal peristaltic contractions (mean(s.e.m.) 82.2(11) versus 39.1(13.8) per cent for satisfactory and poor results respectively; P < 0.05). CONCLUSION: In this retrospective study pneumatic dilatation was effective and safe in nine of 16 patients with dysphagia following antireflux surgery. Peristalsis was normal in eight patients, in seven of whom it was associated with satisfactory results. PMID- 10460653 TI - Electron microscopy in tumour diagnosis: continuing to complement other diagnostic techniques. AB - The histopathological diagnosis of tumours has been transformed by immunohistochemistry. Used with experience and judgement, a panel of antibodies or antisera, combined when necessary with antigen retrieval, will enable the accurate typing of most problematic tumours. This has led many histopathologists to question whether the electron microscope has any residual utility for tumour diagnosis; the machines are large, costly to purchase and maintain, and will accept only minute samples of tissue. The following articles by Mierau and by Eyden, both strong advocates, comment on the current and future role of electron microscopy in tumour diagnosis. PMID- 10460654 TI - Angiofibroblastoma of the skin: a histological, immunohistochemical and ultrastructural report of two cases of an undescribed fibrous tumour. AB - AIMS: The aim of this report is to present two cases of a distinct mesenchymal tumour of the skin that does not fit into one of the established entities. METHODS AND RESULTS: All cases of fibromyxoid tumours from the files of two dermatopathology centres, together with clinical data and histopathological records, were reviewed. Two cases of a lesion composed of stellate and spindle shaped cells with the phenotype of fibroblasts embedded in a fibromyxoid to dense fibrous stroma were identified. Because of the large number of capillary-sized blood vessels and their peculiar distribution within the stroma, the name angiofibroblastoma of the skin is proposed for this peculiar neoplasm. In both cases, the tumour appeared as an indolent and slowly growing nodule on the extremities of adults. Immunohistochemical and, in one case, ultrastructural studies were performed. CONCLUSIONS: The lack of recurrences during a follow-up period of 9 and 3 years, respectively, after complete excision together with the bland histopathological appearance supports the benign character of the tumour. PMID- 10460655 TI - Intraneural angiosarcoma and angiosarcoma arising in benign and malignant peripheral nerve sheath tumours: clinicopathological and immunohistochemical analysis of four cases. AB - AIMS: Angiosarcomatous differentiation represents the least common form of heterologous differentiation in malignant peripheral nerve sheath tumours (MPNST), and is seen most frequently in patients with neurofibromatosis type 1. More rarely, it has been reported in patients without stigmata of neurofibromatosis, or in benign nerve sheath tumours and peripheral nerves. This study was undertaken to confirm this rare association. METHODS AND RESULTS: Four cases of angiosarcoma arising in a peripheral nerve, in a long-standing schwannoma and in two MPNST are described. Immunohistochemical studies were performed on paraffin sections with the alkaline phosphatase-antialkaline phosphatase method. An intraneural high-grade epithelioid angiosarcoma arose in the left posterior tibial nerve of a 78-year-old man, a well to moderately differentiated angiosarcoma was seen in an ancient schwannoma of the lateral neck in a 73-year-old women, and an angiosarcoma of varying grades of differentiation developed in a recurrent MPNST in the thigh of 38-year-old man. In addition a high-grade MPNST in the axillary region of a 30-year-old man showed foci of heterologous high-grade angiosarcomatous differentiation. The neural and endothelial lines of differentiation were confirmed in each case by positive immunohistochemical staining for neural and endothelial markers, respectively. In all cases tested, the neural differentiated cells stained immunohistochemically positive for antibodies against vascular endothelial growth factor. CONCLUSIONS: This study confirms the rare association of angiosarcoma arising in peripheral nerves, as well as in benign and malignant peripheral nerve sheath tumours. PMID- 10460656 TI - High mitotic index associated with poor prognosis in gastrointestinal autonomic nerve tumour. AB - AIMS: Three gastrointestinal autonomic nerve tumours (GANT) were characterized by immunohistochemistry and flow cytometry. Two of the three cases occurred in the small intestine, while the third was found in the stomach. Besides the immunohistochemical and ultrastructural description, the aim of this study was to examine the relation between the known and accepted predictive factors (ploidy data, the S-phase fraction, the mitotic and MIB-1 index and the size of the tumour) and the survival of the patients. METHODS AND RESULTS: The immune profile showed that 3/3 cases were vimentin and NSE, 2/3 were synaptophysin and PGP 9.5 positive, while 1/3 also showed S100 positivity. Ultrastructurally, all the cases had dense core granules, one of them contained skenoid fibres. The flow cytometry revealed diploid DNA in all cases, however, significant differences could be seen in the proliferative activity of the individual neoplasms. CONCLUSIONS: In spite of the published data of gastrointestinal stromal tumours (GIST) generally, neither the MIB-1 index and the ploidy data nor the size of the primary tumour helped to predict the clinical progression of the examined GANTs. However, the high proliferative activity (57 mitoses/10 HPF) and the elevated S-phase fraction (24%) was associated with advanced, metastatic and recurring disease in case 3. On the basis of these three cases, high mitotic activity is the most reliable factor in predicting aggressive clinical behaviour. PMID- 10460657 TI - A novel immunohistochemical detection system using mirror image complementary antibodies (MICA). AB - AIMS: To describe and illustrate a novel and highly sensitive peroxidase-based immunohistochemical detection system which employs mutually attractive, mirror image complementary antibodies (MICA). METHODS AND RESULTS: To demonstrate the sensitivity of the MICA system alongside the avidin-biotin complex (ABC) method, we selected a range of mouse monoclonal and rabbit polyclonal primary antibodies against antigens that are generally regarded as relatively difficult or impossible to detect on formalin-fixed, paraffin-embedded lymphoid tissue. Compared with the ABC method, the MICA immunodetection method enabled us to dilute primary antibodies up to 200-fold with equivalent or superior immunostaining results and, usually, considerably shortened primary antibody incubation times. CONCLUSIONS: We have described and illustrated a novel immunohistochemical detection system and demonstrated greatly increased sensitivity over the commonly used ABC system. An additional advantage of the MICA system is that it is avidin-free and so avoids non-specific staining due to endogenous tissue biotin. PMID- 10460658 TI - Successful application of indirect in-situ polymerase chain reaction to tissues fixed in Bouin's solution. AB - AIMS: To evaluate the value of polymerase chain reaction-in situ hybridization (PCR-ISH) for the detection of human papillomaviruses (HPV) in paraffin sections of cervical biopsies fixed either in 10% formalin or in Bouin's solution. METHODS AND RESULTS: We analysed 40 biopsies from Italian women infected with the human immunodeficiency virus type 1 (HIV 1). In-situ hybridization techniques were performed with commercial biotinylated probes. The PCR-ISH was carried out by the 'hot start modification'. Cervical intraepithelial neoplasia (CIN) was found in 23 of 40 patients (57. 5%); eight cases showed condylomatous features. Human papillomavirus was detected in 42.5% by ISH and in 65% by PCR-ISH. Sixty-nine per cent of positive biopsies contained HPV 16, 18, 31 and 33. HPV 6 and 11 were found only in condylomata acuminata samples. CONCLUSIONS: The results point to a high incidence of HPV infection as well as of CIN in HIV-positive patients. Human papillomavirus type 16 appears to be most frequently associated with CIN. Polymerase chain reaction-ISH is more sensitive than ISH in the detection and typing of HPV DNA both in clinical and in 'latent' infections. The two techniques yielded the same results with either formalin- or Bouin's-fixed material. PMID- 10460659 TI - CD44v6 expression in inflammatory bowel disease is associated with activity detected by endoscopy and pathological features. AB - AIMS: Overexpression of CD44v6 in colon crypt epithelial cells has been suggested to have diagnostic potential in differentiating ulcerative colitis from other forms of colon inflammation, including Crohn's disease. Our aim was to determine the value of CD44v6 expression in inflammatory bowel disease (IBD) and to look for possible associations between CD44v6 expression and activity of this disease. METHODS AND RESULTS: CD44v6 expression was studied using immunohistochemical techniques in 100 surgical and endoscopic colon samples of ulcerative colitis (n = 71) and Crohn's disease (n = 29), and in every case disease activity was studied by endoscopy and microscopic examination. Fifty-five of 71 (77.5%) samples of ulcerative colitis showed monoclonal antibody 2F10 stained colon epithelium, as did 16 of 29 (55.2%) samples of Crohn's disease. CD44v6 was detected in 88.2% (15 of 17) of cases of IBD with severe disease activity and in 100% of eight cases of severe ulcerative colitis. Our study showed a strong association between CD44v6 expression and the activity of IBD (P = 0.007). CONCLUSIONS: CD44v6 expression in IBD is significantly associated with activity detected by means of endoscopy and pathological features. Our data suggest that CD44v6 expression may have some usefulness in conjunction with other factors as a means of evaluating the disease activity. Moreover, CD44v6 expression was higher in ulcerative colitis than Crohn's disease (P = 0.02), although this does not confirm the utility of monoclonal antibody 2F10 in differential diagnosis between ulcerative colitis and Crohn's disease, as there was a notable percentage of positive samples of Crohn's disease. PMID- 10460660 TI - Small cell carcinoma of urinary bladder is differentiated from urothelial carcinoma by chromogranin expression, absence of CD44 variant 6 expression, a unique pattern of cytokeratin expression, and more intense gamma-enolase expression. AB - AIMS: Small cell (neuroendocrine) carcinoma of the urinary bladder is clinically more aggressive than urothelial (transitional cell) carcinoma. We have investigated the immunohistochemical markers most useful in diagnosing small cell carcinoma in bladder. METHODS AND RESULTS: We evaluated the expression of chromogranin A, CD44 variant 6 (CD44v6), cytokeratin (CAM 5.2), gamma-enolase, synaptophysin, and CD45 in 46 small cell carcinomas of the bladder. Small cell and urothelial carcinoma were mixed in 21 (46%) cases. The two immunohistochemical markers with best ability to discriminate between small cell and urothelial carcinoma were chromogranin A and CD44v6. Chromogranin A had 97% specificity for small cell carcinoma, staining 65% of cases with 2+/3+ mean intensity; only one case (5%) of urothelial carcinoma was weakly (1+/3+) positive. CD44v6 was 80% specific for urothelial carcinoma, with immunoreactivity in 60% of cases, compared with 7% of small cell carcinoma cases. In cases positive for CD44v6, the mean percentage of reactive urothelial carcinoma cells was 75% (range 10-100%), greater than the 12% of cells in three cases of small cell carcinoma (P = 0.31); further, the pattern of immunoreactivity was membranous vs. focal cytoplasmic, respectively. All small cell carcinomas stained with one of the three neuroendocrine markers tested; 76% of cases were reactive for synaptophysin and 93% for gamma-enolase, with specificities of 86% and 73% in comparison to urothelial carcinoma. gamma-enolase staining of small cell carcinoma was more intense (P = 0.01) than for urothelial carcinoma. Cytokeratin CAM 5.2 stained a mean 47% of cells in small cell carcinoma, always in a punctate perinuclear pattern, and 75% in urothelial carcinoma, in a membranous pattern. CONCLUSIONS: CD44v6, chromogranin A, and possibly gamma-enolase and cytokeratin (CAM 5.2) help differentiate small cell carcinoma from urothelial carcinoma. PMID- 10460661 TI - Papillary renal cell carcinoma with clear cell cytomorphology and chromosomal loss of 3p. AB - AIMS: Cytogenetic studies on renal cell carcinomas (RCCs) have disclosed a correlation between chromosome aberrations and histomorphological features. Nevertheless, it is still controversial whether the cytomorphology of the tumour cells (clear cell, chromophilic, chromophobe) or their growth pattern (nonpapillary, papillary) is more discriminative for the combined histomorphological-cytogenetic classification of RCCs. METHODS AND RESULTS: Three RCCs with papillary growth pattern and clear cell cytomorphology were analysed by classical cytogenetics using standard G-banding techniques. Each tumour displayed clonal aberrations leading to loss of terminal 3p chromosomal segments. Monosomy 14 was also consistently found. Trisomy 17 was not observed in any of the tumours. CONCLUSIONS: This series of three RCCs consisting of clear cells with papillary architecture revealed chromosomal aberrations characteristic for the conventional (clear cell) RCC. Irrespective of the predominant papillary growth pattern, none of the cases were characterized by trisomy of chromosomes 3q, 7, 8, 12, 16, 17 and 20 and loss of Y chromosome which are widely regarded as the most consistent genetic alterations for papillary RCC. Therefore, our cytogenetic findings provide evidence that papillary clear cell RCCs should be classified according to their cytomorphology rather than their growth pattern even when papillary architecture is prominent. PMID- 10460662 TI - p53 expression in phyllodes tumours is associated with histological features of malignancy but does not predict outcome. AB - AIMS: To study p53 protein expression in phyllodes tumours of the breast, with particular attention to its prevalence and to its relationship with histological features and clinical outcome. METHODS AND RESULTS: Stromal and epithelial p53 immunohistochemical expression was studied in 57 phyllodes tumours (27 benign, 17 borderline, 13 malignant) using an avidin-biotin peroxidase method. High levels of expression (> 30% of stromal nuclei) were found in eight phyllodes tumours (14%). p53 expression was associated with tumour grade (P = 0.001), prominent stromal overgrowth (P = 0.0003), prominent stromal nuclear pleomorphism (P = 0.006), high stromal mitotic count (P = 0.05), and an infiltrative tumour margin (P = 0. 05). Six patients were lost to follow-up after surgery. Mean follow-up time of the remaining 51 patients was 7.3 years (median 4. 3, range 0.5-25) or until death. Sixteen patients (31%) experienced tumour recurrence. Recurrence was more likely if there was an infiltrative tumour margin (P = 0.006) or prominent stromal overgrowth (P = 0.04) but not p53 expression (P = 0.55). A minority of recurrences expressed p53 more extensively than their primary counterparts. There were five tumour-related deaths (10% of patients). Death was associated with high grade (P = 0.0002), prominent stromal overgrowth (P = 0.0001), an infiltrative margin (P = 0.0002), prominent nuclear pleomorphism (P = 0.005), a high mitotic count (P = 0.01) and tumour size (P = 0.03). Again, p53 expression was not associated with tumour-related survival (P = 0. 13). CONCLUSIONS: p53 abnormalities occur in a minority of borderline and malignant phyllodes tumours. p53 expression is associated with known negative prognostic factors, but does not appear to be a useful determinant of tumour recurrence or long-term survival. PMID- 10460663 TI - Glioblastoma multiforme in a mature ovarian teratoma with recurring brain tumours. AB - AIMS: We report a case study to highlight the occurrence of glioblastoma multiforme in an ovarian teratoma. METHODS AND RESULTS: A 10-year-old girl presented with a left frontal lobe primitive neuroectodermal tumour which was successfully treated. After 6 uneventful years, she developed glioblastoma multiforme located posterior to the site of the initial tumour. Six years later, she presented with a mature cystic teratoma containing glioblastoma multiforme. CONCLUSIONS: Glioblastoma in an ovarian teratoma is an exceptional event, which might have an initial clinical presentation as a metastatic brain tumour. Alternatively, recurring glial tumours may occur in a genetically predisposed person; the role of radiation and chemotherapy in this context remains to be elucidated. PMID- 10460664 TI - Apoptosis in the endometrium. PMID- 10460665 TI - Ljubljana classification of epithelial hyperplastic laryngeal lesions. PMID- 10460666 TI - Mammary fibroadenoma in a male-to-female transsexual. PMID- 10460667 TI - Congenital lipoid adrenal hyperplasia. PMID- 10460669 TI - Amyloid tumour (amyloidoma) of the leg: histology, immunohistochemistry and electron microscopy. PMID- 10460668 TI - Large B-cell lymphoma of the mandible comprising filiform and signet-ring cells. PMID- 10460670 TI - Chondrolipoma of the breast presenting as calcifications in a routine mammogram. PMID- 10460671 TI - Second-order stereology and ultrastructural examination of the spatial arrangements of tissue compartments within glomeruli of normal and diabetic kidneys. AB - The present study explores 3D spatial arrangements of compartments within the rat renal glomerulus and tests for differences after chemically-induced diabetes. In particular, the arrangements of capillaries, podocytes, mesangium and urinary space are quantified and compared between (a) kidneys within groups and (b) kidneys from streptozotocin-diabetic rats and age-matched controls. The stereological tool employed is the pair correlation function which is estimated by counting linear dipole probes of different sizes superimposed on ultrathin sections so as to be random in position and orientation. Unbiased estimates of the volume density of each glomerular component were estimated by point counting. Thereafter, estimates of the covariance and pair correlation function were determined from corresponding dipole counts. Plots of covariance and pair correlation functions against dipole length were almost identical in control and diabetic groups, indicating that diabetes did not disturb the normal spatial arrangements within glomeruli. However, differences were detected between compartments within groups. Whilst volume elements within all compartments were clustered at distances below about 8 microm (the approximate size of the basic cellular or other structural unit), the cluster size varied between compartments. The pattern was one of progressively smaller clusters in the sequence capillaries, podocytes, urinary space, mesangium. Beyond a distance of 8 microm, all glomerular components (in both control and diabetic groups) were arranged as expected for a 'random' (meaning neither clustered nor repulsed) volume process. These studies re-emphasize the relative invariance of biological organization and the value and limitations of covariance analysis for quantifying different levels of organization in different tissues and experimental groups. PMID- 10460673 TI - Secondary phase distribution analysis via finite body tessellation AB - The concept of a Dirichlet tessellation has been extended to that of a 'finite body' tessellation to provide a more meaningful description of the spatial distribution of non-spherical secondary phase bodies on two-dimensional sections. A finite body tessellation consists of a network of cells constructed from the interfaces of each individual secondary phase body such that every point within a cell is closer to the corresponding body than to any other. Spatial distribution related cell characteristics derived from Dirichlet tessellations have been extended to finite body tessellations. Quantitative comparisons between the two methods indicate that finite body tessellation measurements are more physically representative as well as more sensitive to local distribution characteristics of secondary phases. To reflect the potential application of finite body tessellations, a methodology is described for analysing the effects of particle distribution and morphology on short crack behaviour in particulate reinforced metal matrix composites. PMID- 10460672 TI - Stereological estimation of covariance using linear dipole probes. AB - Classical stereology is capable of quantifying the total amount or 'density' of a geometrical feature from sampled information, but gives no information about the local spatial arrangement of the feature. However, stereological methods also exist for quantifying the 'local' spatial architecture of a 3D microstructure from sampled information. These methods are capable of quantifying, in a statistical manner, the spatial interaction in a structure over a range of distances. One of the key quantities used in a second-order analysis of a volumetric feature is the set covariance. Previous applications of covariance analysis have been 'model-based' and relied upon computerized image analysis. In this paper we describe a new 'design-based' manual method, known as linear dipole probes, that is suitable for estimating covariance from microscopic images. The approach is illustrated in practice on vertically sectioned lung tissue. We find that only relatively sparse sampling per animal is required to obtain estimates of covariance that have low inter-animal variability. PMID- 10460674 TI - Stereological analysis and modelling of gradient structures AB - Gradient structures are inhomogeneous along a particular gradient direction but homogeneous perpendicular to that direction. Consequently, structural parameters such as volume fraction or surface area density are local characteristics which depend on the 'vertical' coordinate with respect to the 'vertical' gradient axis. Analogously, models for gradient structures have model parameters depending on the vertical coordinate z. For example, a Voronoi tessellation with a gradient is generated by a gradient point process with a local intensity which is a function of z. Similarly, a gradient germ grain model is obtained from a gradient point process where the grain size distribution may also depend on z. For a gradient Boolean model, local volume fraction VV(z) and local surface area density SV(z) can be calculated from the model parameters. Stereological methods for gradient structures are based on vertical sections parallel to the gradient direction. Estimation of VV(z), SV(z) and local length density LV(z) is done by lineal analysis using horizontal test lines with vertical coordinate z. Similarly, lineal analysis is used to estimate local mean cell volume of gradient tessellations. For the estimation of local particle number density and size in the spirit of the Wicksell problem the use of kernel methods and distributional assumptions is required. PMID- 10460675 TI - Single-fluorophore imaging with an unmodified epifluorescence microscope and conventional video camera AB - Single fluorophores in aqueous solution were imaged in real time with a conventional silicon-intensified target video camera connected to an unmodified commercial microscope (IX70, Olympus) with epifluorescence excitation with a high pressure mercury lamp. Neither a powerful laser nor an extremely sensitive video camera was required. Three experimental systems were used to demonstrate quantitatively that individual, moving or stationary Cy3 fluorophores could be imaged with the microscope: Cy3-gelsolin attached to an actin filament sliding over heavy meromyosin, sliding actin filaments sparsely labelled with Cy3, and heavy meromyosin labelled with one or two Cy3 fluorophores. The results should encourage many laboratories to attempt 'single-molecule physiology' in which the functions and mechanisms of molecular machines are studied at the single-molecule level in an environment where the biological machines are fully active. PMID- 10460676 TI - Resolution beyond the rayleigh limit using beam displacement AB - Two fluorescence microscope systems which claim to achieve resolution beyond the Rayleigh limit have recently been described. These systems operate using two displaced beams focused on the sample and produce the image from the region of overlap. We describe an analogous system, which performs in a similar manner but does not rely on fluorescence. The imaging performance of these systems is analysed and we show that they all give improved resolution although, crucially, the optical bandwidth is not increased. These systems merely attenuate the lower spatial frequencies and, although such systems can be useful and operate in a manner analogous to pupil plane filters, it is important to appreciate that they do not offer true superresolution, contrary to the impression given in previous papers. PMID- 10460677 TI - Localization of lead in Allium cepa L. cells by electron microscopy. AB - The study of mechanisms by which plants tolerate lead requires ultrastructural observations of lead distribution in cells. However, the conventional technique used in electron microscopy brings up the problem of lead translocation from tissues during chemical processing. If most of the lead is washed out of tissues, then the method is not suitable for this type of study; if, however, it remains in the tissues, the method can be used. The amount of lead washed out of Allium cepa root tips during successive stages of fixing and dehydrating was determined in this study. Allium cepa plants were treated with Pb(NO3)2 in hydroponic cultures. The conventional fixing (GA, OsO4) and dehydration (alcohol + propylene oxide or acetone) methods used in the preparation of tissues for electron microscopy were then applied to root tips. The lead content in tissues and in reagents was analysed in the successive stages of the procedure. It was shown that 96.2% of the lead taken up was retained in the tissues and was not washed out during fixing and dehydrating. Of the 3.8% of the lead lost in the chemical procedure, about half was washed out during fixing in osmium tetroxide. This reagent is thus the least suitable for this type of study. Our study showed that the conventional electron microscopy preparative technique is appropriate for studying the distribution of lead deposits in A. cepa root cells, owing to the way in which lead is compartmentalized/sequestered in plant cells. PMID- 10460678 TI - Backscattered electron imaging of titanium dioxide in frozen hydrated preparations AB - Backscattered electron (BSE) imaging was used to study ultrafine TiO2 crystals distribution in a test cream. The cream was fast frozen, cryofractured and observed uncoated at low temperature. The BSE detector was a microchannel plate. The results demonstrate that up-to-date photoprotective preparations can be investigated by this technique. PMID- 10460679 TI - Effect of anticoagulation and lavage prior to casting of postmortem material with Mercox and Batson 17. AB - In this study we compare the quality of vascular casts, obtained from organs of several animal species from different sources and sacrificed under different conditions. Organs from healthy animals were injected with two different polymers such as Mercox and Batson No. 17. When the specimens were observed under a scanning electron microscope structural elements such as endothelial nuclear impressions on vessels and capillaries, endothelial cell borders, venous valves, imprints of smooth muscle cells and intra-arterial cushions were identified. Organs excised post mortem from large animals can be used for microvascular corrosion casting studies with optimal results. PMID- 10460681 TI - Introduction. PMID- 10460680 TI - Combined ultramicrotomy for AFM and TEM using a novel sample holder. AB - A new sample holder that allows combined microtomy for atomic force microscopy (AFM) and transmission electron microscopy (TEM) is described. The main feature of this sample holder is a small central part holding the sample. This central part fits into the head of an atomic force microscope. AFM measurements can be performed with a sample mounted in this central part of the sample holder. This makes the alignment of a microtomed bulk sample unnecessary, and offers the opportunity of an easy and fast combined sample preparation for AFM and TEM. PMID- 10460682 TI - Quantitative metallography by electron backscattered diffraction. AB - Although electron backscattered diffraction (EBSD) in the scanning electron microscope is used mainly to investigate the relationship between local textures and microstructures, the technique has now developed to the stage where it requires serious consideration as a tool for routine quantitative characterization of microstructures. This paper examines the application of EBSD to the characterization of phase distributions, grain and subgrain structures and also textures. Comparisons are made with the standard methods of quantitative metallography and it is shown that in many cases EBSD can produce more accurate and detailed measurements than the standard methods and that the data may sometimes be obtained more rapidly. The factors which currently limit the use of EBSD for quantitative microstructural characterization, including the speed of data acquisition and the angular and spatial resolutions, are discussed, and future developments are considered. PMID- 10460683 TI - A comparison of grain imaging and measurement using horizontal orientation and colour orientation contrast imaging, electron backscatter pattern and optical methods. AB - The problems associated with the definition of a grain, grain size measurement, and the issues associated with making one- and two-dimensional measurements on a three-dimensional structure are discussed. The relatively new scanning electron microscope (SEM)-based techniques of colour orientation contrast imaging (COCI) and automated electron backscatter pattern (EBSP) are explained and examples given. Comparisons with conventional (horizontal) orientation contrast imaging (HOCI) in the SEM are made. A direct comparison is made between conventional metallographic methods and these new techniques on precisely the same region of an interstitial free iron specimen. Both optical imaging and HOCI were found to miss a large number of grain boundaries (7 and 12%, respectively), and to create boundaries ( approximately 2%). COCI was found to be reliable, with only 3% of boundaries missed. EBSP was taken to be the standard against which the others were compared, as it unambiguously measured changes in crystallographic orientation. Errors in the number of grain boundaries that are detected have a considerable effect on grain size measurements, e.g. mean linear intercept, and a follow-on effect on the modelling of mechanical properties. New methods for increasing the acquisition speed of orientation maps are discussed, along with examples. The combination of COCI (for grain location) and EBSP (for orientation measurement) is promising, but requires improvements in either imaging or image analysis to be totally reliable. PMID- 10460684 TI - Study of dislocation structures near fatigue cracks using electron channelling contrast imaging technique (ECCI). AB - The fatigue of copper single crystals, orientated for single slip, has been studied using electron channelling contrast imaging in a scanning electron microscope. With the incident beam set at the Bragg condition, changes in the backscattered electron intensity occur as the beam is scanned over dislocations that cause a local tilting of the diffraction planes. This technique allows the evolution of dislocation structures over large areas to be followed through different stages of the fatigue life. Furthermore, it enables direct imaging of dislocation configurations at crack tips. The technique is compared with transmission electron microscopy and electron backscatter diffraction in its application to fatigue studies. PMID- 10460685 TI - Source point calibration from an arbitrary electron backscattering pattern. AB - Precise knowledge of the position of the source point is a requirement if electron backscattering patterns (EBSPs) are to be used for crystal orientation measurements or other types of measurements which demand a geometrical analysis of the patterns. Today, possibly the most popular method for locating the source point is a computational technique which uses the positions of a number of indexed Kikuchi bands for calculating the coordinates of the point. A serious limitation of this calibration technique is, however, that the localized bands must first be indexed, which is difficult if the location of the source point is not known with reasonable precision. This paper describes a new technique which determines the location of the source point from the positions of a number of bands in an arbitrary EBSP. Besides the positions of the Kikuchi bands, the only information which is required by this new calibration procedure is the same crystallographic information which is used for normal indexing of EBSPs. The procedure is shown to work successfully with patterns from a simple cubic crystal, as well as with patterns from an orthorhombic BiSCCO superconductor. In the former case, four bands are shown to be sufficient to ensure a unique determination of the source point, whereas five bands are required in the latter case. Once the bands have been localized, the time required for calculating the source point position is of the order of 1 min on a standard PC. PMID- 10460686 TI - Electron backscatter diffraction of grain and subgrain structures - resolution considerations. AB - Characterization of microstructures containing small grains or low-angle grain boundaries by electron backscattered diffraction (EBSD) is limited by the spatial and angular resolution limits of the technique. It was found that the best effective spatial resolution (60 nm) for aluminium alloys in a tungsten-filament scanning electron microscope (SEM) was obtained for an intermediate probe current which provided a compromise between pattern quality and specimen interaction volume. The same specimens and EBSD equipment when used with a field-emission gun SEM showed an improvement in spatial resolution by a factor of 2-3. For characterizing low-angle boundary microstructures, the precision of determining relative orientations is a limiting factor. It was found that the orientation noise was directly related to the probe current and this was interpreted in terms of the effect of probe current on the quality of the diffraction patterns. PMID- 10460687 TI - Problems in determining the misorientation axes, for small angular misorientations, using electron backscatter diffraction in the SEM. AB - The errors associated with calculating misorientation axes from electron backscatter diffraction (EBSD) data have been assessed experimentally. EBSD measurements were made on the same grains after imposed rotations of 2 degrees, 5 degrees, 7 degrees, 10 degrees, 12 degrees, 17 degrees, 27 degrees and 180 degrees around the normal to the specimen surface. The misorientation magnitudes and the misorientation axes associated with the imposed rotations have been calculated from the EBSD data. Individual measurements of misorientation axes are precise for misorientation magnitudes greater than approximately 20 degrees. The errors must be appreciated when assessing misorientation data at lower misorientation magnitudes and particularly at magnitudes less than 5 degrees. Where misorientation axes can be characterized by the distribution of axes from a number of individual measurements, current EBSD techniques are satisfactory, for data sets of 30 measurements, as long as misorientation magnitudes are 10 degrees or more. With larger data sets it may be possible to extend this approach to smaller misorientation magnitudes. For characterization of individual misorientations less than 5 degrees, new EBSD techniques need to be developed. PMID- 10460688 TI - Crystallographic analysis of facets using electron backscatter diffraction. AB - Applications of electron backscatter diffraction (EBSD), also known as backscatter Kikuchi diffraction in the scanning electron microscope (SEM) are first and foremost microtexture and grain boundary misorientation analysis on a single polished section in the specimen. A more subtle and revealing approach to analysis of these data is to use EBSD to probe the orientations of planar surfaces, i.e. facets, which bound crystals. These surfaces include: * grain or phase boundaries * fractures * cracks It is of great interest to know the crystallography of such facets since it provides a key to understanding the physical properties of them. As far as investigation methodology is concerned, surfaces or facets associated with polycrystals are of two types: exposed or unexposed. Exposed facets, such as a fracture surface, can be viewed directly in the SEM, whereas unexposed facets, such as a grain boundary, are usually revealed as an etched trace on a polished surface. Photogrammetric methods can be used to obtain the positional orientation of an exposed facet, and the crystallographic orientation is obtained either directly from the surface or by indirect sectioning. Calibrated sectioning is required to obtain the equivalent parameters for an internal surface. The present paper compares the methods for obtaining and interpreting the crystallography of facets, with illustrations from several materials. PMID- 10460689 TI - Electron backscatter diffraction: applications for nuclear materials. AB - The diffraction of electrons was first observed in 1928 by Kikuchi. The phenomenon results in the formation of characteristic diagrams of the crystalline lattice and the orientation of the phase. Backscattered electrons are diffracted by the different crystallographic planes (hkl) according to the Bragg angle thetab. These describe, by symmetry, two cones of axes normal to the diffracting plane. Information is collected on a phosphor screen, leading to the acquisition of a diffraction pattern called a Kikuchi diagram. Several improvements now give a wide range of applications such as phase identification (carbides or complex compounds in a (U,Zr,O) structure), analysis of materials interfaces (ZrO2/UO2), as well as solidification studies and local texture determination (molybdenum sheets). In these applications, EBSD, as a type of quantitative metallography, is a powerful tool. PMID- 10460690 TI - Influence of grain orientations on the initiation of fatigue damage in an Al-Li alloy. AB - The variation in microstructure and texture in a rectangular bar extruded from a billet of spray-cast 8090 Al-Li alloy has been examined. The fine grain size of the as sprayed billet and the moderate extrusion ratio ( approximately 25 : 1) were seen to cause geometric dynamic recrystallization (GDR) in regions of higher strain towards the edge of the bar. The grain morphology varied from the expected elongated grains at the centre of the bar to equiaxed grains where GDR occurred at the bar edges. A <111> + <100> double fibre texture, significantly distorted towards rolling components and varying through the bar thickness, was found using electron backscatter diffraction. Fatigue resulted in a high density of short secondary cracks, many of which had arrested at grain boundaries. The cracks preferentially nucleated in grains from the <100> fibre texture corresponding to high Schmid factors. PMID- 10460691 TI - Mushrooms, tumors, and immunity. AB - Medicinal properties have been attributed to mushrooms for thousands of years. Mushroom extracts are widely sold as nutritional supplements and touted as beneficial for health. Yet, there has not been a critical review attempting to integrate their nutraceutical potential with basic science. Relatively few studies are available on the biologic effects of mushroom consumption, and those have been performed exclusively in murine models. In this paper, we review existing data on the mechanism of whole mushrooms and isolated mushroom compounds, in particular (1-->3)-beta-D-glucans, and the means by which they modulate the immune system and potentially exert tumor-inhibitory effects. We believe that the antitumor mechanisms of several species of whole mushrooms as well as of polysaccharides isolated from Lentinus edodes, Schizophyllum commune, Grifola frondosa, and Sclerotinia sclerotiorum are mediated largely by T cells and macrophages. Despite the structural and functional similarities of these glucans, they differ in their effectiveness against specific tumors and in their ability to elicit various cellular responses, particularly cytokine expression and production. Unfortunately, our data base on the involvement of these important mediators is still rather limited, as are studies concerning the molecular mechanisms of the interactions of glucans with their target cells. As long as it remains unclear what receptors are involved in, and what downstream events are triggered by, the binding of these glucans to their target cells, it will be difficult to make further progress in understanding not only their antitumor mechanisms but also their other biological activities. PMID- 10460692 TI - Isoprenoid-mediated inhibition of mevalonate synthesis: potential application to cancer. AB - Pure and mixed isoprenoid end products of plant mevalonate metabolism trigger actions that suppress 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase activity. These actions modulate HMG CoA reductase mRNA translation and the proteolytic degradation of HMG CoA reductase. Such post-transcriptional events, we propose, are activated directly by acyclic isoprenoids and indirectly by cyclic isoprenoids. Isoprenoids, acting secondarily to the dominant transcriptional effector of sterologenesis, modestly lower cholesterol levels, if and only if, sterologenesis is not repressed by a saturating imput of dietary cholesterol. An anomaly associated with tumor growth-a sterol feedback-resistant HMG CoA reductase activity-ensures a pool of sterologenic pathway intermediates. Such intermediates provide lipophilic anchors essential for membrane attachment and biological activity of growth hormone receptors, nuclear lamins A and B, and oncogenic ras. Tumor HMG CoA reductase retains high sensitivity to the isoprenoid mediated secondary regulation. Repression of mevalonate synthesis by plant derived isoprenoids reduces ras and lamin B processing, arrests cells in G1, and initiates cellular apoptosis. This unique tumor cell-specific sensitivity allows isoprenoids to be used for tumor therapy, an application emulating that of the statins, but one free of adverse effects. When evaluated at levels provided by a typical diet, isoprenoids individually have no impact on cholesterol synthesis and tumor growth. Nonetheless, isoprenoid-mediated activities are additive, and, sometimes synergistic. Therefore, the combined actions of the estimated 23,000 isoprenoid constituents of plant materials, acting in concert with other chemopreventive phytochemicals, may explain the lowered cancer risk associated with a diet rich in plant products. In contrast, that lowering of cancer risk does not correspond to supplemental intake of other dietary factors associated with fruits, vegetables, and cereal grains, namely fiber, beta-carotene, vitamin C, and vitamin E, and only weakly to supplemental folate. PMID- 10460693 TI - Endothelin-1 and endothelin receptor antagonists in cardiovascular remodeling. AB - Endothelins build a peptide family composed of three isoforms, each of them containing 21 amino acids. Endothelin-1 is the isoform mainly responsible for any cardiovascular action and therefore the sole scope of this review. Endothelin-1 is the most potent endogenous vasoconstrictor known; in addition it acts as a potent (co)mitogen. There is a substantial body of experimental evidence that endothelin-1 may contribute not only to sustained vasoconstriction, but also to remodeling within the cardiovascular system. Thus, with the help of endothelin receptor antagonists (available for a few years) the involvement of mainly ETA receptors in structural diseases such as heart failure, pulmonary hypertension, atherosclerosis, restenosis, systemic hypertension, and chronic renal failure has been shown. These data make endothelin receptor antagonists, and especially those selective for the ETA receptor, promising agents for the treatment of chronic cardiovascular diseases associated with remodeling. Currently several chemically distinct, orally available members of this novel class of therapeutic agents are under clinical investigation. PMID- 10460694 TI - Food restriction-like effects of dehydroepiandrosterone: decreased lymphocyte numbers and functions with increased apoptosis. AB - Both dietary dehydroepiandrosterone (DHEA) and food restriction can prevent or modulate the initiation or progression of a number of diseases in rodents and prolong life span. We sought to determine if these interventions have common mechanisms of action in regulating lymphocyte functions and cell numbers. We observed that male C57BL/6 mice receiving DHEA in the diet (0.45%, w/w) ate approximately 50% as much food as mice on the DHEA-free diet, and this was reflected in decreased body weights throughout a 10-week period. Mice either fed the DHEA-containing diet or pair-fed to the DHEA-treated mice had decreased spleen and thymus weights and lymphocyte cell numbers compared to mice having free access to the control diet. Mice were fed these diets for 2 weeks before and 2 weeks after exposure to sublethal irradiation (500 cGy). In mice fed DHEA or pair-fed, there was a decrease in spleen cell numbers, and B cells were the most severely affected. The frequency of apoptosis in peripheral blood cells increased from <5% in nonirradiated controls to >50% within 4 days after starting DHEA or pair-feeding. Shortly after irradiation, >87% of blood lymphocytes were hypodiploid (apoptotic) in all groups. By 9 days only 27% of lymphocytes in mice on the control diet were hypodiploid compared to 62% in DHEA and 74% in pair-fed mice. In addition, both DHEA and pair-fed mice had significant reductions in T cell function (contact hypersensitivity to dinitrofluorobenzene), B cell function (antibody response to trinitrophenolated-lipopolysaccharide), and NK cell function (lung clearance of radiolabeled YAC-1 tumor cells) 2 weeks after irradiation. In a complementary study, peripheral blood lymphocytes from naive mice were treated overnight with various concentrations of either DHEA or hydrocortisone 21-acetate. Only glucocorticoid-treated cells underwent apoptosis. Thus, DHEA induces apoptosis in vivo but not in vitro. We conclude that dietary DHEA induces apoptosis and decreased lymphocyte production and function in C57BL/6 mice largely by reducing food intake. PMID- 10460695 TI - Effect of pregnancy, postnatal growth, and gender on renal sulfate transport. AB - Serum sulfate concentrations are increased in infants, young children, and pregnant women, compared with adult values. The objective of this investigation was to examine the influences of age, gender, and pregnancy on renal sulfate transport using guinea pigs as an animal model. Membrane vesicles were isolated from the kidney cortex of male animals at four different ages, from male and female adult animals, and from pregnant and nonpregnant female animals. There were no significant differences in marker enzymes for the brush-border membrane (BBM) or basolateral membrane (BLM) among all groups examined. Uptake was determined by a rapid filtration method and membrane fluidity by measuring the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene. The Vmax values for Na+ /sulfate co-transport in BBM were significantly increased with decreasing age, whereas the Km for this process was unchanged. The Vmax and Km for Na + /sulfate co-transport in BBM of pregnant animals were significantly higher than the values in the nonpregnant group. Bicarbonate-driven anion exchange of sulfate in BLM was not different among the different age groups. The Vmax for the bicarbonate/sulfate exchange process in BLM was not different between pregnant and nonpregnant groups; however, the Km for this process in BLM of pregnant animals was significantly greater than the value in nonpregnant animals. There were no gender-related differences in sulfate transport in BBM or BLM isolated from adult male and female animals. Renal BBM fluidity was increased with decreasing age and in pregnant animals, suggesting that altered membrane fluidity may represent one possible mechanism to explain the increased sodium/sulfate uptake in young and pregnant animals. The higher Vmax for Na+/sulfate co-transport in young and pregnant animals suggests that there is an increased density of co-transporter protein or an increase in the rate of movement of the carrier protein (i.e., turnover) once loaded with sodium and sulfate. This increased conservation of inorganic sulfate in young and pregnant guinea pigs may be related to the increased demand for sulfated substrates, such as sulfated glycosaminoglycans, during growth and development. PMID- 10460696 TI - Prolactin and growth hormone stimulation of lactation in mice requires thyroid hormones. AB - This experiment tested the hypothesis that thyroid hormones are essential for a milk production response to growth hormone (GH) and prolactin (PRL). Prior to breeding, female transgenic mice expressing the herpes simplex type-I thymidine kinase in the thyroid were treated with ganciclovir to ablate thyroid follicular cells. To provide for normal gestation, thyrocyte-ablated mice were supplied thyroxine (T4) in drinking water (0.2 microgram/ml) until 7 days before parturition. Litter size was adjusted to 9 pups, hormone administration began on Day 2 of lactation, and mice were sacrificed on Day 12. There were 5-6 mice in each of 7 treatments that included nonablated controls, thyrocyte-ablated controls, and thyrocyte-ablated mice treated with T4, GH, PRL, GH + T4, and PRL + T4. Thyroxine was administered in drinking water, and GH and PRL (20 microgram/d) were administered by subcutaneous injection. Compared with thyrocyte-ablated controls, litter weight gain was unaffected when dams were treated with GH, PRL, or T4 alone. However, when dams were treated with GH or PRL in combination with T4, litter weight gain increased 13% compared with thyrocyte-ablated controls and 18% compared with GH or PRL-treated mice. Concentration of T4 in serum of pups averaged 62 ng/ml and did not differ among treatments. Concentration of T4 in serum of dams averaged 76 ng/ml when T4-treated. Thyroxine 5'-deiodinase (5'D), the enzyme that converts T4 to triiodothyronine, was quantitated in liver, kidney, and mammary gland. Quantity of 5'D was lower in liver and kidney of thyrocyte-ablated dams without T4 than in respective tissues of mice treated with T4, and there was no effect of GH or PRL. However, in mammary gland, 5'D was increased by treatment with GH, PRL, or T4. Data show that thyroid hormones are necessary for a galactopoietic response to GH and PRL and demonstrate a unique organ-specific regulation of 5'D by galactopoietic hormones. PMID- 10460697 TI - A study of the metabolism of apolipoprotein B100 in relation to insulin resistance in African American males. AB - The purpose of this study was to determine the relationship between insulin resistance and apoB100 metabolism in African American males. Fifteen subjects, 33 +/- 7.6 years old, were divided into two groups, insulin-resistant (IR) or insulin-sensitive (IS), based on the sum of the plasma insulin concentrations during an oral glucose tolerance test. The IR group (n = 8) differed significantly from the IS group (n = 7) with respect to body mass index (BMI) (30.1 vs 23.1 kg/m2; P = 0.0003), fasting triglycerides, (118 vs 54 mg/dl, P = 0. 013), and total plasma apolipoprotein B100 (80 vs 59 mg/dl, P = 0.014). Significantly elevated apoB100 levels in the IR group were seen in very low density lipoprotein (VLDL) (5.1 vs 3.4 mg/dl, P = 0.045) and intermediate density lipoprotein (IDL) (18 vs 12 mg/dl, P = 0.017) but not in low density lipoprotein (LDL) (57 vs 46 mg/dl, P = 0.19). Total cholesterol, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein A-I, and blood pressure were not significantly different between the two groups. There was a high correlation between the sum of insulins during the oral glucose tolerance test and the BMI (rho = 0.88, P = 0.0001). In five IR and five IS subjects, apoB100 kinetics were determined in the fasting state using a bolus dose of deuteroleucine and multicompartmental modeling. IR subjects had significantly lower fractional catabolic rates (FCR) in the larger VLDL1 (-70%), the smaller VLDL2 (-71%), and the IDL (-53%) fractions. No significant differences in production rates were observed for any lipoprotein class. There was a significant correlation between the sum of insulins and the FCR of the apoB100 of VLDL1 (rho = -0.65, P = 0.05) and of IDL (rho = -0.85, P = 0.004). The correlation coefficient of the sum of insulins and the FCR of VLDL2 was -0.61 with P = 0.067. We conclude that in this population of African American males, IR is correlated with a decreased FCR of apoB100 in VLDL and IDL and elevated plasma levels of apoB and triglycerides (TG). These changes might be explained by decreased clearance of the TG-rich lipoproteins. We postulate that this may reflect decreased lipoprotein and/or hepatic lipase activity related to insulin resistance and its association with obesity. PMID- 10460698 TI - A new neurological mutant rat with symmetrical calcification of Purkinje cells in cerebellum. AB - A new neurological mutant has been found in the inbred F344 strain of rats. The mutation is inherited as an autosomal recessive trait and is manifest clinically by a hesitant and wobbling gait with asynergic limbs and slight tremor. These symptoms begin at 16-18 days of age and remain essentially constant thereafter. Histologic examination revealed severe degeneration of the Purkinje cells and symmetrical calcification in these and in their dendritic branches in the cerebellar cortex. Such calcified Purkinje cells were intensely stained with the periodic acid-Schiff (PAS) method. PAS-positive substances in the Purkinje cells and extending diffusely over the lesioned sites in the molecular layer were also evident before calcification took place. We have named this neurological mutant the Cerebellar Calcification (CC) rat with the gene symbol cc. This offers a new animal model for the study of the Purkinje cell degeneration and intracranial calcification. PMID- 10460699 TI - Effect of hamster pregnancy on female protein, a homolog of serum amyloid P component. AB - Pentraxins such as human serum amyloid P component (SAP) and C reactive protein (CRP) represent an ancient family of proteins that are ubiquitous in nature and have evolved with little change in structure or regulation. The pentraxin in the Syrian hamster (Mesocricetus auratus) is unique because it is preferentially expressed in the female at high constitutive levels and accordingly called female protein (FP) or FP(SAP) due to its close homology with human SAP. The high levels of FP in female serum (100-fold greater than male serum) suggested its role in hamster pregnancy, one of the shortest of any eutherian mammal. We determined the serum FP concentration in pregnant Syrian hamsters and found a marked decrease (>80%) at term with the nadir at parturition with subsequent increase. A similar downregulation of FP was found in the normal female Syrian hamster after injury (acute phase response), so in both cases the assumed beneficial effects were achieved with less, rather than more pentraxin, a paradoxical pentraxin response. The fall in serum FP concentration could represent a response to protect the fetus from the high and potentially toxic level of FP normally found in the female, that is harmful because of its association with amyloidosis. An FP that is 97.5% identical to Syrian hamster FP is found in the Turkish hamster (Mesocricetus brandti), although serum levels in females are much lower, and amyloid is very rare. During pregnancy/parturition of Turkish hamsters, the serum level of FP remained remarkably constant. In a more distantly related hamster, the Armenian hamster (Cricetulus migratorius), serum FP actually increased during pregnancy and at parturition in a manner similar to that found in the Armenian hamster during an acute phase response. The heterogeneity of FP kinetics during pregnancy in these three species of hamster indicates pleomorphic gene structure for regulation of their similar FPs, and suggests that this protein may have a different function in the pregnancy of each species. PMID- 10460700 TI - The role of nitric oxide in saline-induced natriuresis and diuresis in rats. AB - This study was designed to determine to what extent nitric oxide (NO) mediates the natriuretic and diuretic responses to acute isotonic saline (0.9 gram % NaCl) volume expansion (SVE, 0.5 ml min-1 kg-1). Studies were performed on 49 pentobarbital anesthetized (65 mg/kg) female Sprague-Dawley rats with or without a NO synthase inhibitor, Nomega-nitro-L-arginine (LNA). Group 1 received saline at 27 microliter/min for 1 hr (baseline) and then SVE for 1 hr; Groups 2-4 received LNA at 10, 150, and 200 microgram kg-1 min-1, respectively, for 1 hr followed by LNA + SVE. To determine to what extent inhibition of NOS would reverse an ongoing SVE-induced natriuresis and diuresis, Group 5 was saline volume-expanded for hours 1 and 2 whereas Group 6 was administered SVE during the first hour and then SVE + 150 microgram kg -1 min-1 LNA during the second hour. SVE caused a significant (P < 0.05) increase in the glomerular filtration rate (GFR) of Group 1 and the LNA-treated rats (Groups 2-4). This SVE-induced increase in the GFR occurred despite the fact that baseline GFR was significantly lower in the two groups of rats that were infused with the highest doses of LNA (Groups 3 4). SVE was also associated with similar increases in urine flow rate, sodium and potassium excretion, and total osmolar excretion in Groups 1-4. On the other hand, mean arterial pressure (MAP) was significantly higher in Group 2 during SVE + LNA and during the baseline as well as during the SVE periods in Groups 3-4; MAP was also significantly elevated in Group 6 during SVE + LNA. Thus, despite the fact that MAP was higher in LNA-treated rats, sodium and urine flow rates were the same as in Group 1 (i.e., there was no evidence of a pressure natriuresis or diuresis in these animals). Along these lines, there was a small but significant positive linear correlation coefficient (r = 0.41, P = 0.05) between sodium excretion values and corresponding MAP values in SVE control rats but not in Groups 3-4 during SVE (r = 0.28, P = 0.26). The current data demonstrate that 1) NO does not mediate SVE-induced hyperfiltration in the rat, 2) NO also does not mediate SVE-induced natriuresis or diuresis, and 3), consistent with other reports, NO appears to mediate pressure natriuresis and diuresis. PMID- 10460701 TI - Reversal of propranolol blockade of adrenergic receptors and related toxicity with drugs that increase cyclic AMP. AB - An overdose of propranolol, a widely used nonselective beta-adrenergic receptor blocking agent, can result in hypotension and bradycardia leading to irreversible shock and death. In addition, the blockade of adrenergic receptors can lead to alterations in neurotransmitter receptors resulting in the interruption of the activity of other second messengers and the ultimate cellular responses. In the present experiment, three agents, aminophylline, amrinone, and forskolin were tested in an attempt to reverse the potential lethal effects of a propranolol overdose in dogs. Twenty-two anesthetized beagle dogs were given a 10-min infusion of propranolol at a dose of 1 mg/kg/min. Six of the dogs, treated only with intravenous saline, served as controls. Within 15-30 min all six control dogs exhibited profound hypotension and severe bradycardia that led to cardiogenic shock and death. Seven dogs were treated with intravenous aminophylline 20 mg/kg 5 min after the end of the propranolol infusion. Within 10 15 min heart rate and systemic arterial blood pressure returned to near control levels, and all seven dogs survived. Intravenous amrinone (2-3 mg/kg) given to five dogs, and forskolin (1-2 mg/kg) given to four dogs, also increased heart rate and systemic arterial blood pressure but the recovery of these parameters was appreciably slower than that seen with aminophylline. All of these animals also survived with no apparent adverse effects. Histopathologic evaluation of the hearts of the dogs treated with aminophylline showed less damage (vacuolization, inflammation, hemorrhage) than the hearts from animals given propranolol alone. Results of this study showed that these three drugs, all of which increase cyclic AMP, are capable of reversing the otherwise lethal effects of a propranolol overdose in dogs. PMID- 10460702 TI - 5-hydroxytryptamine evokes endothelial nitric oxide synthase activation in bovine aortic endothelial cell cultures. AB - Activation of endothelial nitric oxide synthase (eNOS) results in the production of nitric oxide (NO) that mediates the vasorelaxing properties of endothelial cells. The goal of this project was to address the possibility that 5 hydroxytryptamine (5-HT) stimulates eNOS activity in bovine aortic endothelial cell (BAEC) cultures. Here, we tested the hypothesis that 5-HT receptors mediate eNOS activation by measuring agonist-stimulated [3H]L-citrulline ([3H]L-Cit) formation in BAEC cultures. We found that 5-HT stimulated the conversion of [3H]L arginine ([3H]L-Arg) to [3H]L-Cit, indicating eNOS activation. The high affinity 5-HT1B receptor agonist, 5-nonyloxytryptamine (5-NOT)-stimulated [3H]L-Cit turnover responses were concentration-(0.01 nM to 100 microM) and time-dependent. Maximal responses were observed within 10 min following agonist exposures. These responses were effectively blocked by the 5-HT1B receptor antagonist, isamoltane, the 5-HT1B/5-HT2 receptor antagonist, methiothepin, and the eNOS selective antagonists (0.01-10 microM): L-Nomega -monomethyl-L-arginine (L-NMMA) and L-N omega-iminoethyl-L-ornithine (L-NIO). Pretreatment of BAEC cultures with pertussis toxin (PTX; 1-100 ng/ml) for 16 hr resulted in significant inhibition of the agonist-stimulated eNOS activity, indicating the involvement of Gi proteins. These findings lend evidence of a 5-HT1B receptor/eNOS pathway, accounting in part for the activation of eNOS by 5-HT. Further investigation is needed to determine the role of other vascular 5-HT receptors in the stimulation of eNOS activity. PMID- 10460704 TI - Occult hyperextension "toddler's" fracture by Swischuk, et al. PMID- 10460703 TI - Guidelines of care for laser surgery. American Academy of Dermatology. Guidelines/Outcomes Committee. PMID- 10460705 TI - Elective tracheal intubation and the intubating laryngeal mask. PMID- 10460706 TI - Difficult laryngoscopy and difficult intubation. PMID- 10460707 TI - Perception of pain experienced and adequacy of analgesia following elective craniotomy. PMID- 10460708 TI - Why mothers die. PMID- 10460709 TI - Blood culture in apyrexial patient having epidural blood patch. PMID- 10460710 TI - Rapid sequence induction. PMID- 10460711 TI - 'Target'-controlled infusion: misnamed and misplaced. PMID- 10460712 TI - Ultrasound guided central vein cannulation. PMID- 10460713 TI - Pitfalls of disposable equipment. PMID- 10460714 TI - Care, cars and the millennium bug. PMID- 10460715 TI - Use of a cuffed oropharyngeal airway and Aintree catheter in a difficult airway. PMID- 10460716 TI - Laryngoscopy grades. PMID- 10460717 TI - Epidural protocol audit. PMID- 10460718 TI - Labelling of obstetric patients. PMID- 10460719 TI - Pulsating mass in the neck following attempted internal jugular vein catheterisation. PMID- 10460720 TI - Treating Guillain-Barre syndrome by filtration of cerebrospinal fluid. PMID- 10460721 TI - Forearm venous pressures and blood back flow. PMID- 10460722 TI - An unfortunate cause of death. PMID- 10460723 TI - Raynaud's phenomenon and propofol. PMID- 10460724 TI - Epidural anaesthesia in the anaesthetised patient. PMID- 10460725 TI - A plan for the NHS for the 21st century. PMID- 10460726 TI - Scissors as a propofol ampoule 'snapper'? PMID- 10460727 TI - Precordial stethoscopes in children. PMID- 10460729 TI - Locomotor forces on a swimming fish: three-dimensional vortex wake dynamics quantified using digital particle image velocimetry. AB - Quantifying the locomotor forces experienced by swimming fishes represents a significant challenge because direct measurements of force applied to the aquatic medium are not feasible. However, using the technique of digital particle image velocimetry (DPIV), it is possible to quantify the effect of fish fins on water movement and hence to estimate momentum transfer from the animal to the fluid. We used DPIV to visualize water flow in the wake of the pectoral fins of bluegill sunfish (Lepomis macrochirus) swimming at speeds of 0.5-1.5 L s(-)(1), where L is total body length. Velocity fields quantified in three perpendicular planes in the wake of the fins allowed three-dimensional reconstruction of downstream vortex structures. At low swimming speed (0.5 L s(-)(1)), vorticity is shed by each fin during the downstroke and stroke reversal to generate discrete, roughly symmetrical, vortex rings of near-uniform circulation with a central jet of high velocity flow. At and above the maximum sustainable labriform swimming speed of 1.0 L s(-)(1), additional vorticity appears on the upstroke, indicating the production of linked pairs of rings by each fin. Fluid velocity measured in the vicinity of the fin indicates that substantial spanwise flow during the downstroke may occur as vortex rings are formed. The forces exerted by the fins on the water in three dimensions were calculated from vortex ring orientation and momentum. Mean wake-derived thrust (11.1 mN) and lift (3.2 mN) forces produced by both fins per stride at 0.5 L s(-)(1) were found to match closely empirically determined counter-forces of body drag and weight. Medially directed reaction forces were unexpectedly large, averaging 125 % of the thrust force for each fin. Such large inward forces and a deep body that isolates left- and right-side vortex rings are predicted to aid maneuverability. The observed force balance indicates that DPIV can be used to measure accurately large-scale vorticity in the wake of swimming fishes and is therefore a valuable means of studying unsteady flows produced by animals moving through fluids. PMID- 10460728 TI - Pseudotuberculous pyelonephritis: a rare entity and a diagnostic pitfall. PMID- 10460730 TI - Locomotion in sturgeon: function of the pectoral fins. AB - Pectoral fins are one of the major features of locomotor design in ray-finned fishes and exhibit a well-documented phylogenetic transition from basal to derived clades. In percomorph fishes, the pectoral fins are often used to generate propulsive force via oscillatory movements, and pectoral fin propulsion in this relatively derived clade has been analyzed extensively. However, in the plesiomorphic pectoral fin condition, exemplified by sturgeon, pectoral fins extend laterally from the body in a generally horizontal orientation, have been assumed to generate lift to balance lift forces and moments produced by the heterocercal tail, and are not oscillated to generate propulsive force. The proposal that pectoral fins in fishes such as sturgeon generate lift during horizontal locomotion has never been tested experimentally in freely swimming fishes. In this paper, we examine the function of pectoral fins in sturgeon swimming at speeds from 0.5-3.0 L s(-)(1), where L is total body length. Sturgeon were studied during steady horizontal locomotion as well as while sinking and rising in the water column. Pectoral fin function was quantified using three dimensional kinematics to measure the orientation of the fin surface, digital particle image velocimetry (DPIV) was used to describe flow in the wake of the fin and to estimate force exerted on the water, and electromyography was used to assess pectoral fin muscle function. Sturgeon (size range 25-32 cm total length) swam horizontally using continuous undulations of the body with a positive body angle that decreased from a mean of 20 degrees at 0.5 L s(-)(1) to 0 degrees at 3.0 L s(-)(1). Both the angle of the body and the pectoral fin surface angle changed significantly when sturgeon moved vertically in the water column. Three dimensional kinematic analysis showed that during steady horizontal swimming the pectoral fins are oriented with a negative angle of attack predicted to generate no significant lift. This result was confirmed by DPIV analysis of the pectoral fin wake, which only revealed fin vortices, and hence force generation, during maneuvering. The orientation of the pectoral fins estimated by a two-dimensional analysis alone is greatly in error and may have contributed to previous suggestions that the pectoral fins are oriented to generate lift. Combined electromyographic and kinematic data showed that the posterior half of the pectoral fin is actively moved as a flap to reorient the head and body to initiate rising and sinking movements. A new force balance for swimming sturgeon is proposed for steady swimming and vertical maneuvering. During steady locomotion, the pectoral fins generate no lift and the positive body angle to the flow is used both to generate lift and to balance moments around the center of mass. To initiate rising or sinking, the posterior portion of the pectoral fins is actively moved ventrally or dorsally, respectively, initiating a starting vortex that, in turn, induces a pitching moment reorienting the body in the flow. Adjustments to body angle initiated by the pectoral fins serve as the primary means by which moments are balanced. PMID- 10460731 TI - Tail kinematics of the chub mackerel Scomber japonicus: testing the homocercal tail model of fish propulsion. AB - Scombrid fishes possess a homocercal caudal fin with reduced intrinsic musculature and dorso-ventrally symmetrical external and internal morphology. Because of this symmetrical morphology, it has often been assumed that scombrid caudal fins function as predicted by the homocercal tail model. According to that model, the caudal fin moves in a dorso-ventrally symmetrical manner and produces no vertical lift during steady swimming. To test this hypothesis, we examined the tail kinematics of chub mackerel, Scomber japonicus (24.8+/-1.3 cm total length, L). Markers were placed on the caudal fin to identify specific regions of the tail, and swimming chub mackerel were videotaped from lateral and posterior views, allowing a three-dimensional analysis of tail motion. Analysis of tail kinematics suggests that, at a range of swimming speeds (1.2-3.0 L s(-)(1)), the dorsal lobe of the tail undergoes a 15 % greater lateral excursion than does the ventral lobe. Lateral excursion of the dorsal tail-tip also increases significantly by 32 % over this range of speeds, indicating a substantial increase in tail-beat amplitude with speed. In addition, if the tail were functioning in a dorso-ventrally symmetrical manner, the tail should subtend an angle of 90 degrees relative to the frontal (or xz) plane throughout the tail beat. Three-dimensional kinematic analyses reveal that the caudal fin actually reaches a minimum xz angle of 79.8 degrees. In addition, there is no difference between the angle subtended by the caudal peduncle (which is anterior to the intrinsic tail musculature) and that subtended by the posterior lobes of the tail. Thus, asymmetrical movements of the tail are apparently generated by the axial musculature and transmitted posteriorly to the caudal fin. These results suggest that the caudal fin of the chub mackerel is not functioning symmetrically according to the homocercal model and could produce upward lift during steady swimming. PMID- 10460732 TI - Antibody to H(+) V-ATPase subunit E colocalizes with portasomes in alkaline larval midgut of a freshwater mosquito (Aedes aegypti). AB - The pH profile, gross structure, ultrastructure and immunolabeling of the mosquito (Aedes aegypti) larval midgut are described as a first step in analyzing the role of plasma membrane H(+ )V-ATPase in the alkalization of the gut, nutrient uptake and ionic regulation. Binding of an antibody to H(+ )V-ATPase subunit E colocalizes with 'portasomes' (approximately 10 nm in diameter), which are thought to correspond to the V(1) part of the H(+) V-ATPase. In gastric caeca (pH 8), both antibody-binding sites and portasomes are located apically; in the anterior midgut (pH 10-11), they are located basally; and in the posterior midgut (pH approximately equal to 8) they are again located apically. The hypothesis that the energization of alkalization is mediated by an H(+) V-ATPase is supported by the inability of larvae to maintain the high pH after 72 h in 10 (micro)M bafilomycin B1. Confirming earlier reports, the two principal epithelial cell types are designated as 'columnar' and 'cuboidal' cells. The apical plasma membranes (microvilli) of epithelial cells in the gastric caeca and basal infoldings of anterior midgut are invaded by mitochondria that lie within approximately 20 nm of the portasome-studded plasma membranes. The colocalization of V-ATPase-immunolabeling sites and portasomes to specific plasma membranes within so-called 'mitochondria-rich' cells of gastric caeca and anterior midgut suggests that midgut alkalization in mosquitoes is achieved by molecular mechanisms similar to those that have been described in caterpillars, even though the gross structure of the midgut and the localization of the V-ATPase are dissimilar in the two species. In caterpillars, the high alkalinity is thought to break down dietary tannins, which block nutrient absorption; it may play a similar role in plant-detritus-feeding mosquito larvae. The colocalization of immunolabeling sites and portasomes, together with the presence of long, 'absorptive-type' microvilli in the posterior midgut, suggest that the V-ATPase energizes nutrient uptake there. PMID- 10460734 TI - Metamorphosis in the summer flounder Paralichthys dentatus: changes in gill mitochondria-rich cells. AB - Salinity tolerance changes during larval development and metamorphosis in the summer flounder (Paralichthys dentatus) and other teleosts. The physiological mechanisms responsible for osmoregulation during these early stages of development are not well understood. This study characterized changes in ultrastructure, intracellular membranes and immunoreactive Na(+)/K(+)-ATPase of mitochondria-rich cells (MRCs) in the gills of summer flounder during metamorphosis. Gill ultrastructure at the start of metamorphosis revealed only one type of MRC, which had weak reactivity to osmium and lacked a well-defined apical pit. In juveniles, two types of MRCs were observed: light-staining MRCs (LMRCs) with weak reactivity to osmium, and dark-staining MRCs (DMRCs) with strong reactivity to osmium and positioned adjacent to LMRCs. Compared with MRCs at the start of metamorphosis, the mitochondria of juvenile MRCs appeared smaller, with more transverse cristae and electron-dense matrices. Changes in MRCs during metamorphosis were also accompanied by increased immunoreactive Na(+)/K(+)-ATPase. These findings suggest that gill MRCs develop during the metamorphosis of summer flounder as the gill takes on an increasingly important osmoregulatory role. PMID- 10460733 TI - Contraction parameters, myosin composition and metabolic enzymes of the skeletal muscles of the etruscan shrew Suncus etruscus and of the common European white toothed shrew Crocidura russula (Insectivora: soricidae). AB - In the Etruscan shrew, the isometric twitch contraction times of extensor digitorum longus (EDL) and soleus muscles are shorter than in any other mammal, allowing these muscles to contract at outstandingly high contraction frequencies. This species has the highest mass-specific metabolic rate of all mammals and requires fast skeletal muscles not only for locomotion but also for effective heat production and for an extremely high ventilation rate. No differences could be detected in the fibre type pattern, the myosin heavy and light chain composition, or in the activity of the metabolic enzymes lactate dehydrogenase and citrate synthase of the two limb muscles, the EDL and the soleus, which in larger mammalian species exhibit distinct differences in contractile proteins and metabolic enzymes. All properties determined in EDL and soleus muscles of Suncus etruscus, as well as in the larger Crocidura russula, are typical for fast oxidative fibres, and the same holds for several other skeletal muscles including the diaphragm muscle of S. etruscus. Nevertheless, the EDL and soleus muscles showed different mechanical properties in the two shrew species. Relaxation times and, in C. russula, time to peak force are shorter in the EDL than in the soleus muscle. This is in accordance with the time course of the Ca(2+) transients in these muscles. Such a result could be due to different parvalbumin concentrations, to a different volume fraction of the sarcoplasmic reticulum in the two muscles or to different Ca(2+)-ATPase activities. Alternatively, the lower content of cytosolic creatine kinase (CK) in the soleus compared with the EDL muscle could indicate that the observed difference in contraction times between these shrew muscles is due to the CK-controlled activity of their sarcoplasmic reticulum Ca(2+)-ATPase. PMID- 10460735 TI - Leptocephalus energetics: metabolism and excretion. AB - Leptocephali are the unusual transparent larvae that are typical of eels, bonefish, tarpon and ladyfish. Unlike the larvae of all other fishes, leptocephali may remain in the plankton as larvae for several months before metamorphosing into the juvenile form. During their planktonic phase, leptocephali accumulate energy reserves in the form of glycosaminoglycans, which are then expended to fuel metamorphosis. The leptocephalus developmental strategy is thus fundamentally different from that exhibited in all other fishes in two respects: it is far longer in duration and energy reserves are accumulated. It was anticipated that the unusual character of leptocephalus development would be reflected in the energy budget of the larva. This study describes the allocation of energy to metabolism and excretion, two important elements of the energy budget. Metabolic rates were measured directly in four species of leptocephali, Paraconger caudilimbatus, Ariosoma balearicum, Gymnothorax saxicola and Ophichthus gomesii, using sealed-jar respirometry at sea. Direct measurements of metabolic rates were corroborated by measuring activities of lactate dehydrogenase and citrate synthase, two key enzymes of intermediary metabolism, in addition to that of Na(+)/K(+)-ATPase, a ubiquitous ion pump important in osmotic regulation. Excretion rates were determined by subsampling the sea water used in the respiratory incubations. The entire premetamorphic size range for each species was used in all assays. Mass-specific oxygen consumption rate, excretion rate and all enzyme activities (y) declined precipitously with increasing mass (M) according to the equation y=aM(b), where a is a species specific constant and -1.740.40). The narrow-sense genetic correlation between wheel-running and body mass after 8 weeks of wheel access was estimated to be -0. 50. A negative genetic correlation could account for the negative relationship between voluntary wheel-running and body mass that has been reported across 13 species of muroid rodents. PMID- 10460739 TI - Regulation of swimming in the Culex pipiens (Diptera, Culicidae) pupa: kinematics and locomotory trajectories. AB - High-speed videography was used to investigate swimming kinematics and locomotory trajectories during escape responses in the pupa of Culex pipiens (Diptera, Culicidae). The pupa can perform straight-line motion despite undergoing backward and forward somersaults through 400 degrees at a rate of 20 s(-)(1). For linear motion to occur, the effective stroke of the abdomen must be delivered during the part of each cycle when the body is facing forwards, allowing passive rotation to carry the body round to the correct position for the start of the next half stroke. On-off motion of the abdomen is regulated by a click mechanism based on stressed integumentary plates which buckle at strategic points in the cycle. The importance of self-inertia as a component in positional regulation was demonstrated by increasing the kinematic viscosity of the medium: this retarded rotation and resulted in somersaulting on the spot. Whereas normally the pupa invariably directs its swimming downwards, reversal of the usual light gradient produced upward paths as well. It is concluded that straight-line motion is generated automatically by the locomotory machine without any need for sensory feedback about body orientation during each cycle, but that swimming direction can be influenced by sensory cues such as light. PMID- 10460740 TI - Ca(2+) uptake in the sarcoplasmic reticulum from the systemic heart of octopod cephalopods. AB - We have measured Ca(2+) uptake in crude homogenates of heart tissue, as well as cell shortening and ionic currents in isolated myocytes exposed to caffeine, to characterize Ca(2+) uptake in the sarcoplasmic reticulum (SR) of the systemic heart of octopus. The maximal rate of SR Ca(2+) uptake in crude homogenates of octopus heart was 43+/-4 (mean +/- s.e.m., N=7), compared with 28+/-2 nmol min( )(1 )mg(-)(1) protein (N=4) in homogenates of rat heart. The Ca(2+)-dependency of SR Ca(2+) uptake was similar for the two species, with a Ca(2+) activity at half maximal uptake rate (pCa(50)) of 6.04+/-0.02 for octopus and 6.02+/-0.05 for rat. Exposure of isolated myocytes to 10 mmol l(-)(1) caffeine resulted in cell shortening to 53+/-2 % of the resting cell length and an inward trans-sarcolemmal ionic current. The charge carried by this current was 3.28+/-0.70 pC pF(-)(1) (mean +/- s.e.m., N=5) corresponding to extrusion of 34.0+/-0.7 amol Ca(2+ )pF( )(1) from the cell by Na(+)/Ca(2+) exchange. This is approximately 50 times more than the Ca(2+) carried by the Ca(2+) current elicited by a 200 ms depolarization from -80 to 0 mV and corresponds to an increase in the total intracellular [Ca(2+)] of 404+/-86 (&mgr;)mol l(-)(1) non-mitochondrial volume due to Ca(2+) release from the SR. Thus, we find that at 20 degrees C in the SR both Ca(2+) content and Ca(2+) uptake rate in the systemic heart of octopus are comparable with or larger than the corresponding values obtained in the rat heart. These results support the argument that the SR may play an important role in the regulation of contraction in the systemic heart of cephalopods. PMID- 10460741 TI - Circadian oscillation of sensitivity of spider eyes: diurnal and nocturnal spiders. AB - The circadian oscillation of sensitivity of the anterior median eye of a nocturnal spider Araneus ventricosus and that of a diurnal spider Menemerus confusus were examined by recording electroretinograms. The anterior median eye of Araneus ventricosus showed a marked circadian oscillation of sensitivity, but that of Menemerus confusus showed no circadian oscillation. PMID- 10460742 TI - Ultrastructural and protein analysis of surfactant in the Australian lungfish Neoceratodus forsteri: evidence for conservation of composition for 300 million years. AB - The Australian lungfish Neoceratodus forsteri is the most primitive member of the lungfish family, with a surfactant lipid composition similar to the actinopterygiian fishes, which evolved 400 million years ago. We have analysed the proteins associated with surfactant isolated from lung lavage of this species, and used electron microscopy and immunohistochemistry to examine the surfactant structures and the subcellular localisation of these proteins. The epithelial lining of the gas-exchange region of the lungfish lung consists of one basic cell type, which has characteristics of both mammalian alveolar type I and type II cells and may be the common ancestor of both. It has long cytoplasmic plates containing microvilli, large osmiophilic bodies resembling mammalian lamellar bodies and a cytoplasm rich in metabolic organelles. Extracellular structures reminiscent of mammalian surfactant forms, but not including tubular myelin, were observed in the airspaces. Immunochemical analysis of the lungfish surfactant and lung tissue, using antibodies to human SP-A and SP-B, showed a similar staining pattern to human surfactant, indicating that SP-A- and SP-B-like proteins are present. Immunohistochemistry revealed that both SP-A and SP-B reactivity was present in the secretory cell osmiophilic bodies. In conclusion, our results suggest that, despite the great diversity in present day lung structures, a common cellular mechanism may have evolved to overcome fundamental problems associated with air-breathing. PMID- 10460743 TI - Work-dependent deactivation of a crustacean muscle. AB - Active shortening of respiratory muscle L2B from the crab Carcinus maenas results in contractile deactivation, seen as (1) a decline of force during the course of isovelocity shortening, (2) a reduction in the rate of force redevelopment following shortening, (3) a depression of the level of isometric force reached following shortening, and (4) an accelerated relaxation at the end of stimulation. The degree of deactivation increases with increasing distance of shortening, decreases with increasing shortening velocity, and is approximately linearly related to the work done during shortening. Deactivation lasts many seconds if stimulation is maintained, but is largely although not completely removed if the stimulation is temporarily interrupted so that the force drops towards the resting level. Deactivation for a given distance and velocity of shortening increases with increasing muscle length above the optimum length for force production. Stimulating muscle L2B at suboptimal frequencies gives tetanic contractions that are fully fused but of less than maximal amplitude. The depression of force following shortening, relative to the force during an isometric contraction, is independent of the stimulus frequency used to activate the muscle, indicating that deactivation is not a function of the background level of stimulus-controlled muscle activation upon which it occurs. Deactivation reduces the work required to restretch a muscle after it has shortened, but it also lowers the force and therefore the work done during shortening. The net effect of deactivation on work output over a full shortening/lengthening cycle is unknown. PMID- 10460744 TI - Eotaxin. An essential mediator of eosinophil trafficking into mucosal tissues. PMID- 10460745 TI - Exhaled nitric oxide (NO), NO synthase activity, and regulation of nuclear factor (NF)-kappaB. PMID- 10460746 TI - The macroimportance of the pulmonary immune microenvironment. PMID- 10460747 TI - Interleukin (IL)-5 downregulates tumor necrosis factor (TNF)-induced eotaxin messenger RNA (mRNA) expression in eosinophils. Induction of eotaxin mRNA by TNF and IL-5 in eosinophils. AB - An eotaxin is a chemoattractant specific for eosinophils that are known to play a role in helminth infection and allergic responses. Although several cellular sources have been reported to produce eotaxin, it would be interesting to know whether eosinophils are able to produce their own eotaxin and participate in recruitment of themselves in response to inflammation. To this end, a cloned eotaxin complementary DNA was transcribed in vitro to use as a probe for detecting eotaxin messenger RNA (mRNA), and eotaxin protein levels were determined by enzyme-linked immunosorbent assay. Eotaxin mRNA was, as analyzed by in situ hybridization, rarely detectable in unstimulated eosinophils, but was strongly induced in eosinophils when stimulated with tumor necrosis factor (TNF). Interleukin (IL)-5, which is known to be a major factor of eosinophil survival in vivo and in vitro, was also able to induce a modest level of eotaxin mRNA but inhibited TNF-induced eotaxin mRNA expression in a dose-response manner. Dexamethasone inhibited TNF-induced eotaxin mRNA expression. This result was consistent with that from reverse transcription/polymerase chain reaction followed by Southern blot analysis. Unlike the little expression of eotaxin mRNA in the absence of stimuli, the measurement of eotaxin protein revealed that a considerable amount of eotaxin protein was constitutively produced in unstimulated eosinophils. Its expression was upregulated by TNF and IL-5 as well. However, the inhibitory effect of IL-5 on TNF-mediated eotaxin protein production was not as pronounced as that on eotaxin mRNA induction. Collectively, these data reflect the complex physiology of eosinophils in the expression of eotaxin gene upon the exposure to their survival and/or death factors. PMID- 10460748 TI - Nitric oxide blocks nuclear factor-kappaB activation in alveolar macrophages. AB - Nitric oxide (NO) is an important endogenous regulatory molecule implicated in both proinflammatory and antiinflammatory processes in the lung. Previously, we demonstrated that in human alveolar macrophages (AM), NO decreased inflammatory cytokine production, including that of interleukin-1beta, tumor necrosis factor alpha and macrophage inflammatory protein-1alpha. One mechanism by which NO could regulate such diverse cytokine production is through effects on the transcription factor nuclear factor-kappaB (NF-kappaB), which controls the expression of the genes for these inflammatory cytokines and growth factors. We therefore investigated whether NO affects NF-kappaB activation in AM in vitro and in vivo. In vitro studies with AM showed that NF-kappaB activation by lipopolysaccharide (LPS) is decreased by NO in a dose-dependent manner. NO prevented an LPS-mediated decrease in the NF-kappaB inhibitory protein IkappaB-alpha. In asthma, airway NO levels are increased, whereas in primary pulmonary hypertension (PPH), airway NO levels are lower than in healthy lungs. In vivo investigations were conducted with freshly isolated AM from healthy controls, asthmatic individuals, and PPH patients. Healthy individuals had airway NO levels of 8 +/- 2 ppb (mean +/- SEM), which is associated with low NF-kappaB activation. Asthma patients with airway NO levels > 17 ppb showed minimal NF-kappaB activation, whereas asthmatic individuals with NO levels 0.2). CONCLUSION: After cataract surgery only a minor difference exists between the colour vision scores of diabetic and non diabetic patients. This indicates that accelerated yellowing of the lens in diabetes is the predominant cause of the colour vision anomaly found in phakic diabetic patients. PMID- 10460773 TI - Treatment of retinal tears and lattice degenerations in fellow eyes in high risk patients suffering retinal detachment: a prospective study. AB - BACKGROUND/AIMS: Fellow eye prophylaxis for retinal detachment (RD) is still a controversial issue since opinions are not unanimous regarding the kind of lesions to be treated or the method of treatment. This prospective clinical study aimed to follow the course of vitreoretinal conditions in 150 high risk fellow eyes. METHODS: 150 consecutive patients with unilateral rhegmatogenous RD were included in this study. Inclusion criteria were good explorability of fellow eye retinal periphery and one of the following conditions in the fellow eye-aphakia, pseudophakia with capsulotomy, high myopia (>-6D), contralateral eye to a giant retinal tear. Prophylactic treatment (photocoagulation or scleral buckling) was performed in the presence of retinal tears and lattice degenerations. The state of the vitreous body was determined at the beginning of the study and at the end, when RD occurred. RESULTS: Follow up ranged from 36 to 132 months. 95 fellow eyes were subjected to laser treatment; five eyes underwent prophylactic surgical treatment. Initially, in the treated group posterior vitreous detachment (PVD) was present in 100 eyes (100% of cases), but as a complete PVD only in 42 of them (42%). 10 eyes in the treated group developed RD during the follow up period. In five of these cases the partial PVD had progressed and a retinal tear in a previously healthy area was the cause of the retinal detachment. In the other five eyes RD apparently developed from previously treated lesions. Progression of PVD was evident in four out of these five eyes. The untreated eyes had no visible degenerative lesions. During follow up eight eyes developed RD. These eyes had no PVD at the beginning of the study, but showed a partial PVD at the time of the diagnosis of RD. CONCLUSION: Fellow eyes with pre-existing retinal tears and PVDs can go on to retinal detachment in spite of laser prophylactic treatment. When PVD is not detectable or a partial PVD is present, the progression of posterior vitreous separation can account for retinal tears and RDs arising in formerly healthy areas. PMID- 10460774 TI - Intravitreal dexamethasone in exogenous bacterial endophthalmitis: results of a prospective randomised study. AB - AIM: To evaluate the efficacy of intravitreal dexamethasone co-administered with intravitreal antibiotics along with vitrectomy in the management of exogenous bacterial endophthalmitis. METHODS: In a prospective randomised clinical trial, 63 patients (63 eyes) with suspected bacterial endophthalmitis (postoperative and post-traumatic) were treated with vitrectomy and intravitreal antibiotics and randomised to intravitreal dexamethasone (IOAB with = 29 eyes) and no dexamethasone (IOAB without = 34 eyes). Inflammation score (IS) and visual acuity were measured by two masked observers before surgery, and at 1, 4, and 12 weeks after surgery in both the groups. RESULTS: There was significant reduction (p <0.0001) in IS at 1, 4, and 12 weeks after the surgery in the "IOAB with" group; there was temporary but significant increase (p <0.01) in IS at 1 week in the "IOAB without" group, before decline (p <0.001) of IS at 4 and 12 weeks. The magnitude and relative percentage change in IS between the two groups were found to be significant at 1 (p <0.0001), and 4 (p <0.01) weeks, and not at 12 weeks. The visual acuity at 12 weeks was comparable in both the IOAB with and IOAB without groups. CONCLUSION: Intravitreal dexamethasone helps in early reduction of inflammation in exogenous bacterial endophthalmitis, but has no independent influence on the visual outcome. In selected patients with endophthalmitis where oral corticosteroids cannot be given for medical reasons intravitreal corticosteroids could be beneficial; in other situations they could be complementary to oral corticosteroid therapy. PMID- 10460775 TI - Effect of panretinal photocoagulation on serum levels of laminin in patients with diabetes: a prospective study. AB - BACKGROUND/AIM: Laminin, a major specific non-collagenous glycoprotein of basement membrane, has been proposed as an index of diabetic retinopathy and high serum concentrations have been reported in patients with proliferative diabetic retinopathy. On the other hand, panretinal photocoagulation (PRP) prevents the progression of severe diabetic retinopathy and reverses preretinal neovascularisation. The aim of the study was to investigate the effect of PRP on serum levels of laminin in patients with diabetes. METHODS: 20 patients with diabetes undergoing PRP and 15 patients with mild or moderate non-proliferative diabetic retinopathy in whom a PRP was not performed were included in the study. Serum laminin-P1 (Lam-P1), the largest pepsin resistant fragment of laminin, was determined by radioimmunoassay in each patient before starting PRP and 3 months after it was accomplished. Similarly, a baseline and a 4 month sample were analysed in the non-photocoagulated controls. RESULTS: Serum Lam-P1 concentrations obtained 3 months after PRP were significantly lower when compared with the initial values (1.62 (SD 0.36) U/ml v 1.91 (0.37) U/ml; p <0.001). A decrease of serum levels of Lam-P1 could be seen in all patients. By contrast, in those patients with mild or moderate non-proliferative diabetic retinopathy in whom a PRP was not performed, no significant changes were detected in serum Lam P1 concentrations (1.72 (0.20) U/ml v 1. 74 (0.17); p=0.250). CONCLUSION: PRP decreases serum Lam-P1 levels in patients with severe diabetic retinopathy. Thus, the studies addressed to evaluate the usefulness of Lam-P1 as a marker of diabetic retinopathy should consider previous PRP as an influencing factor. Finally, our results suggest that retinal source of Lam-P1 strongly contributes to serum Lam-P1 in patients with severe diabetic retinopathy. PMID- 10460776 TI - Everting suture correction of lower lid involutional entropion. AB - AIMS: To assess the long term efficacy of everting sutures in the correction of lower lid involutional entropion and to quantify the effect upon lower lid retractor function. METHODS: A prospective single armed clinical trial of 62 eyelids in 57 patients undergoing everting suture correction of involutional entropion. Patients were assessed preoperatively and at 6, 12, 24, and 48 months postoperatively. The main outcome variables were lower lid position and the change in lower lid retractor function. RESULTS: When compared with the non entropic side, the entropic lid had a greater degree of horizontal laxity and poorer lower lid retractor function. These differences however, were not significant. At the conclusion of the study and after a mean follow up period of 31 months, the entropion had recurred in 15% of the patients. There were no treatment failures in the group of five patients with recurrent entropion. The improvement in lower lid retractor function after the insertion of lower lid everting sutures did not reach statistical significance. There was no significant difference between the treatment failure group and the group with a successful outcome with regard to: the degree of horizontal lid laxity or lower lid retractor function present preoperatively; patient age or sex; an earlier history of surgery for entropion. There was neither a demonstrable learning effect nor a significant intersurgeon difference in outcome. The overall 4 year mortality rate was 30%. CONCLUSIONS: The use of everting sutures in the correction of primary or recurrent lower lid involutional entropion is a simple, successful, long lasting, and cost effective procedure. PMID- 10460777 TI - Concentration dependent effects of hydrogen peroxide on lens epithelial cells. AB - AIMS: To evaluate the effects of hydrogen peroxide exposure on the survival and proliferation of cultured lens epithelial cells. METHODS: TOTL-86 cells, a line of rabbit lens epithelial cells, were used. The survival and proliferation of TOTL-86 cells were quantified by a rapid colorimetric assay (MTT assay). To determine the effects of hydrogen peroxide, TOTL-86 cells were exposed to different concentrations of hydrogen peroxide. To determine the effect of cell numbers on the survival and proliferation of TOTL-86 cells at a fixed concentration of hydrogen peroxide, different numbers of cells were plated and exposed to hydrogen peroxide. To determine whether there is a synergistic effect between hydrogen peroxide and EGF, bFGF, PDGF-AA, and insulin, TOTL-86 cells were exposed to hydrogen peroxide combined with one of these growth factors. RESULTS: High levels (1 mM) of hydrogen peroxide killed TOTL-86 cells and sublethal levels (100 microM) suppressed their proliferation. From 1 nM to 1 microM of hydrogen peroxide, there was a dose dependent increase in the cell numbers. The initial seeded cell number dramatically affected the response to hydrogen peroxide. Although growth factors showed no synergistic effects with hydrogen peroxide on proliferation, both EGF and insulin, but not bFGF or PDGF, rescued TOTL-86 cells from the sublethal effect. CONCLUSION: Hydrogen peroxide in cooperation with some growth factors plays an important role in the proliferation of lens epithelial cell. PMID- 10460778 TI - Alpha/beta- and gamma/delta TCR(+) lymphocyte infiltration in necrotising choroidal melanomas. AB - AIM: To detect specific tumour infiltrating T cells (TIL) carrying antigen specific MHC-I restricted receptor genes on necrotising and non-necrotising malignant melanomas and to correlate the findings with clinical data. METHODS: alpha/beta- and gamma/delta- TIL were determined by immunohistochemical staining in melanomas of patients with known follow up of more than 10 years. An antigen retrieval method was used to determine variable genes delta1 and gamma1 on TCR(+) cells by an anti-TCR Vdelta1 and anti-CrgammaM1, and of Valpha and Vbeta TCR(+) by an anti-pan-TCR(+) alpha/beta antibody. RESULTS: Intratumoral TIL were present in 86 of 113 (76.1%) necrotising melanomas (NMM) v 21 of 100 (21%) in non necrotising melanomas (MM); of these, Valpha/beta- TCR(+) cells were present in 52 of 74 (70.3%) TIL harbouring NMM v four of 21 (19%) MM; Vgamma1 in 29 of 74 (39.2%) NMM v two of 21 (10%) MM; and Vdelta1 in 39 of 74 (52.7%) NMM v three of 21 (14%) MM. Extratumoral lymphocytic infiltration was seen in 86 (76.1%) NMM including Valpha/beta TCR(+) cells in 10 (11.6%) cases, v five (5%) MM cases with no Valpha/beta TCR(+) cells detected. Vgamma1 and Vdelta1 TCR(+) cells were not found in extratumoral infiltrates. CONCLUSIONS: In NMM, the median survival was 69.3 (range 6-237) months, 19 of 74 patients (25.7%) survived 5 years, and mortality was associated with advanced stage (p<0.001), patient age (p<0.023), and extent of necrosis (p<0.048). Survival was increased with evidence of Vgamma1 and Vdelta1 TCR(+) cells (p<0.026). PMID- 10460779 TI - IL-4 regulates chemokine production induced by TNF-alpha in keratocytes and corneal epithelial cells. AB - BACKGROUND: Eosinophils are thought to play a major role in the pathogenesis of corneal lesions in ocular allergies. The regulation of chemokine production in corneal cells by the Th2 cytokine, interleukin 4 (IL-4), was examined in order to investigate its role in ocular allergies. METHODS: Pure cultures of human corneal epithelial cells and keratocytes were exposed to tumour necrosis factor alpha (TNF-alpha) and/or IL-4. 24 hours after exposure, culture supernatants were removed and concentrations of IL-8 and RANTES were quantified by ELISA assay. RESULTS: Simultaneous addition of IL-4 inhibited TNF-alpha induced IL-8 production in both corneal epithelial cells and keratocytes. TNF-alpha and IL-4 synergistically stimulated the production of RANTES in keratocytes. CONCLUSION: Differential regulation of chemokine production from corneal cells by IL-4 may play a role in the selective recruitment of predominantly eosinophils to the ocular surface in ocular allergies. PMID- 10460780 TI - Culture and characterisation of epithelial cells from human pterygia. AB - BACKGROUND/AIMS: Pterygia are a common disorder of the ocular surface. The disease represents a chronic fibrovascular and degenerative process thought to originate at the conjunctival-corneal junction, where altered limbal stem cells are proposed to be the cell of origin. Extensive epidemiological evidence exists to implicate ultraviolet B irradiation in the pathogenesis of pterygia. To date no animal or in vitro culture model has been developed to test such an hypothesis. The aim of this study was to establish and characterise a pure population of epithelial cells derived from pterygium tissue. METHODS: Tissue specimens were obtained from patients undergoing pterygium excision. Explants were cultured in either serum free or serum supplemented medium. Primary and passaged cells were processed for light microscopy, analysed by flow cytometry, and characterised immunohistochemically using specific antibodies. RESULTS: In serum free culture, cuboidal cells with typical morphology of epithelial cells migrated from the pterygium explants from 3 days onwards and eventually formed a cohesive monolayer. Passaged cells consisted of 98.4% cytokeratin positive cells and demonstrated immunoreactivity for multiple cytokeratins, including AE1, AE3, AE5, but were negative for AE8. These cells also expressed an epithelial specific antigen, together with vimentin and mucin, as did epithelial cells in sections of pterygia. CONCLUSIONS: A relatively simple method of isolating pterygium epithelial cells has been established. Cultured pterygium epithelial cells are phenotypically and functionally similar to their in vivo counterparts with respect to keratin, vimentin, and mucin expression. In vitro assays using these cells may aid in elucidating the pathogenesis of pterygia. PMID- 10460781 TI - Hypertension and diabetic retinopathy--what's the story? PMID- 10460782 TI - Paediatric neuro-Behcet's disease presenting with optic nerve head swelling. PMID- 10460783 TI - Should diabetic patients be screened for glaucoma? PMID- 10460784 TI - Hertel exophthalmometry: the most appropriate measuring technique. PMID- 10460785 TI - Histological Typing of Tumours of the Eye and its Adnexa. PMID- 10460786 TI - Walsh and Hoyt's Clinical Neuro-Ophthalmology. PMID- 10460788 TI - Manual of Orbital and Lacrimal Surgery. PMID- 10460787 TI - Laser Photocoagulation of the Retina and Choroid. PMID- 10460789 TI - Eye Examination and Refraction. PMID- 10460790 TI - 1998 International Meeting on the Arylamine N-Acetyltransferases: synopsis of the workshop on nomenclature, biochemistry, molecular biology, interspecies comparisons, and role in human disease risk. AB - On October 22-24, 1998, a workshop was held at Kuranda, Queensland, Australia. The purpose of the meeting was to provide a forum for discussion of a number of diverse research areas of the biochemistry and molecular biology of arylamine N acetyltransferases and to foster collaboration among several major groups of investigators around the world. In addition, participants were asked to consider how the nomenclature system for arylamine N-acetyltransferases could be strengthened to cope with the burgeoning number of new alleles discovered in the last 3 years. The full text of all meeting abstracts can be viewed at. PMID- 10460791 TI - Cytochrome CYP sources of N-alkylprotoporphyrin IX after administration of porphyrinogenic xenobiotics to rats. AB - Cytochrome P-450 (CYP) 3A2 and CYP2C11 are sources of 70 and 30%, respectively, of N-vinylprotoporphyrin IX (N-vinylPP) formation after administration of 3 [(arylthio)ethyl]sydnone (TTMS) to rats. Female rats receiving TTMS were pretreated with dexamethasone, which induces CYP3A1 preferentially to CYP3A2. The resulting 12-fold increase in N-vinylPP formation showed that CYP3A1 was also a source of N-vinylPP. Phenobarbital (PB) pretreatment, which induces CYP2B1/2 and 3A1/2 in male rats, increased N-vinylPP formation after TTMS administration. Troleandomycin, a selective CYP3A inhibitor, was unable to decrease TTMS-mediated N-vinylPP formation in PB-treated male rats, indicating that CYP2B1/2 were sources of N-vinylPP. This conclusion was supported by demonstrating a 15-fold increase in TTMSinduced N-vinylPP formation in female rats after CYP2B1/2 induction with PB pretreatment. Allylispropylacetamide (AIA) inactivates rat CYP2B1/2, 2C6, 2C7, 2C11, and 3A1/2. Troleandomycin was unable to decrease N-AIA protoporphyrin IX adduct (N-AIAPP) formation, showing that CYP3A1/2 were not susceptible to AIA-mediated N-alkylation. N-AIAPP formation in females was approximately 30% of that in males, and thus we attribute 30% of N-AIAPP formation in males to the non-gender-specific isozymes (CYP2C6, 2C7, and/or 2B1/2), whereas approximately 70% originates from CYP2C11. PB treatment in female rats resulted in a 5-fold increase in N-AIAPP formation, showing that CYP2B1/2 were also susceptible to N-alkylation mediated by AIA. 1-Aminobenzotriazole elicited formation of equivalent amounts of N'N-aryl bridged protoporphyrin IX in male and female rat liver, demonstrating that nonselective mechanism-based inactivation is accompanied by nonselective conversion of the CYP heme moieties to N'N-aryl bridged protoporphyrin IX. PMID- 10460792 TI - N-acetylation of the heterocyclic amine batracylin by human liver. AB - Batracylin (8-aminoisoindolo[1,2-b]quinazolin-12(10 H)-one; BAT) is a heterocyclic amine that exhibits antitumor activity in a number of in vivo and in vitro models. The acetyl product has been implicated in BAT toxicity in animals, cells, and bacteria. The ability of human N-acetyltransferase (NAT) to form this product was investigated. Nine human liver samples were analyzed for NAT1 and NAT2 genotypes. Seven of the samples possessed at least one NAT1*4 allele. Three samples contained one or more NAT2*4 allele and were classified as rapid acetylators. The remaining six had two alleles associated with the slow phenotype. NAT activities were evaluated with BAT, sulfamethazine (SMZ), a preferential substrate for human NAT2, and p-aminobenzoic acid, a substrate for NAT1. BAT activities in the nine donor samples ranged from 14.9 to 0.56 nmol/min/mg. The mean apparent K(m) values in rapid acetylators for BAT, SMZ, and p-aminobenzoic acid were 6.59 +/- 3.21, 278 +/- 69.4, and 31.2 +/- 12.5 microM, respectively. The apparent K(m) values for slow acetylators did not differ from the rapid acetylator phenotype. However, a significant difference in the apparent V(max) for BAT and SMZ was observed between rapid and slow acetylators. Comparing the apparent intrinsic clearance (V(max)/K(m)) for BAT and SMZ, a significant correlation (r(2) = 0.97, p <.001) was observed. These data demonstrate that BAT N-acetylation is similar to SMZ, and suggests that BAT is a preferential substrate for human NAT2. Thus, rapid acetylators would be more likely to develop toxicity when exposed to this drug. PMID- 10460793 TI - Modulation of rat cytochrome P-450 by an investigational HIV protease inhibitor. AB - Previous studies in vitro have revealed that L-754,394, an HIV protease inhibitor, is a potent suicide inhibitor of cytochrome P-450 enzymes. The present report examines the effect of chronic treatment of L-754,394 on hepatic cytochrome P-450s in adult male rats. L-754,394 was administered orally once a day for 7 days and resulted in significant changes in marker activities. An unusual parabolic (ascending, then descending) profile was observed for testosterone 2beta-/6beta-(CYP 3A1/2-catalyzed) hydroxylase activities during the 7-day treatment with 20 mg/kg L-754,394. These activities, which were elevated 2 fold on day 2, returned to basal levels by day 8. In contrast, testosterone 2alpha-/16alpha-(CYP2C11-catalyzed) hydroxylase activities showed an opposite parabolic (descending, then ascending) profile during the same period, reducing to 40% of control activities on day 4, followed by a rebounding trend. Immunoquantitation of CYP 3A1/2 and 2C11 showed that the expressed protein levels were in parallel with the associated activities. Furthermore, mRNA levels of CYP 3A2 and CYP2C11 showed the same trends as the protein expression of the respective isoforms. These observations show that L-754,394 perturbs the relative abundance of P-450 isoforms in rat liver by affecting the regulation at a pretranslational step. This may further involve a disturbance of hormonal homeostasis. Although serum levels of testosterone did not show a marked change during treatment, thyroxine and triiodothyronine markedly decreased on days 2 and 4, and subsequently increased to basal levels. PMID- 10460795 TI - Metabolism of antitumor hydroxymethylacylfulvene by rat liver cytosol. AB - Acylfulvenes are a potent class of antitumor agents derived from illudin S, a fungal sesquiterpene. Illudin S possesses antitumor activity but has a poor therapeutic index. Acylfulvene is 100-fold less toxic against human lung adenocarcinoma cells than illudin S, but inhibits tumor growth in human xenografts, opposite to illudin S. An analog of acylfulvene, MGI 114 (hydroxymethylacylfulvene), shows much greater efficacy, producing complete tumor regression in xenograft models. MGI 114 is currently in phase II clinical trials. Cytotoxicity of MGI 114, like that of illudin S, is believed to involve both chemical reaction and enzymatic reduction. Enzymatic reduction by a cytosolic NADPH-dependent enzyme (from rat liver) produced an aromatic metabolite similar to that formed from illudin S. However, the reaction occurred more slowly. In addition, four new metabolites were isolated, two hydroxylated derivatives and two in which the primary allylic hydroxyl was replaced by hydride. All retained the reactive centers of the parent MGI 114. PMID- 10460794 TI - Dose-dependent up-regulation of rat pulmonary, renal, and hepatic cytochrome P 450 (CYP) 1A expression by nicotine feeding. AB - In a previous study in which a single 2.5 mg/kg (15.4 micromol/kg) s. c. dose of nicotine effected a transient, lung-specific induction of cytochrome P-450 (CYP) 1A1 in the rat, a dose-response study and assessment of the lung specificity of the induction was limited by toxicity of the acute parenteral nicotine exposure. In the present study, we examined the dose-CYP1A1/2 induction response relationship and the tissue specificity of the induction by orally administered nicotine, which lacks the toxicity of the parenterally administered drug. Nicotine, administered in a nutritionally balanced liquid diet, at a level of 20 (low), 60 (medium), or 200 (high) mg/kg of diet, induced CYP1A1 in the lung and kidney in a dose-dependent manner and in the liver at the high nicotine dose only, whereas CYP1A2 was induced in the liver dose-dependently and in the kidney at the high nicotine dose only. The high nicotine dose up-regulated mRNA level in the three tissues examined, but with the lung being the most responsive to the up regulation. Induction of the CYP1A1-preferential activity ethoxyresorufin O deethylase by the low, medium, and high nicotine diets was 1.9-, 4.9-, and 21.6 fold, respectively, in the lung, 1.4-, 1.7-, and 15.9-fold, respectively, in the kidney, and 1.7-, 2.9-, and 5.1-fold, respectively, in the liver. Similarly, albeit to lower extents, the dietary alkaloid induced the CYP1A2-preferential activity methoxyresorufin O-demethylase in all three tissues dose-dependently. Plasma nicotine concentration correlated neither with the dietary nor intake dose of the alkaloid nor with tissue levels of CYP1A, especially with the high-dose diet. Plasma nicotine levels at which CYP1A induction was maximal were comparable to those reported in smokers, suggesting that nicotine may induce CYP1A1 in humans. PMID- 10460796 TI - Activated sulfonamides are cleaved by glutathione-S-transferases. AB - In preclinical pharmacokinetic studies and in in vitro rat, dog, and human primary hepatocyte incubations, the sulfonamide (-NH-SO(2)-) bond of a potent inhibitor of the HIV-1 protease containing the p-cyanopyridinyl moiety (PNU 109112), undergoes metabolic cleavage to form the corresponding amine metabolite (PNU-143070). Strikingly, a compound, PNU-140690, obtained by substituting the cyanopyridinyl group of PNU-109112 with a trifluoropyridinyl moiety, was stable under the same in vivo and in vitro conditions used for PNU-109112. The apparent "sulfonamidase activity" present in liver was localized to the cytosolic fraction and shown to be an enzyme-mediated reaction requiring reduced glutathione (GSH). The enzyme responsible was purified in a single step on a GSH immobilized gel and was identified as glutathione-S-transferase (GST) by sequence analysis of peptides obtained by tryptic digestion of the purified protein. Moreover, a mixture of GST isoenzymes purified from rat liver, and three recombinant human GST isoforms, A1-1, M1-1, and P1-1, were active toward PNU-109112 sulfonamide cleavage; the three isoforms exhibited differential rates of PNU-109112 cleavage, demonstrating isoenzyme selectivity. PMID- 10460797 TI - Mechanism, structure-activity studies, and potential applications of glutathione S-transferase-catalyzed cleavage of sulfonamides. AB - The mechanism of sulfonamide cleavage of PNU-109112, a potent HIV-1 protease inhibitor, by glutathione-S-transferase (GST) was investigated in the presence of reduced GSH. GST-catalyzed sulfonamide cleavage takes place via the nucleophilic attack of GSH on the pyridine moiety of the substrate with formation of the GS para-CN-pyridinyl conjugate, the corresponding amine, and sulfur dioxide. Structure activity studies with a variety of sulfonamides indicate that an electrophilic center alpha to the sulfonyl group is required for cleavage. Substituents that withdraw electron density from the carbon atom alpha- to the sulfonyl group facilitate nucleophilic attack by the GS(-) thiolate bound to GST. The rate of sulfonamide cleavage is markedly affected by the nature of the electrophilic group; replacement of para-CN by para-CF(3) on the pyridine ring of PNU-109112 confers stability against sulfonamide cleavage. On the other hand, stability of sulfonamides is less dependent on the nature of the amine moiety. These principles can be applied to the synthesis of sulfonamides, labile toward cellular GST, that may serve as prodrugs for release of bioactive amines. Tumors are particularly attractive targets for these sulfonamide prodrugs as GST expression is significantly up-regulated in many cancer cells. Another potential application could be in organic synthesis, where protection of amines as the corresponding activated sulfonamides can be reversed by GST/GSH under mild conditions. PMID- 10460798 TI - Enhancement of cytochrome P-450 3A4 catalytic activities by cytochrome b(5) in bacterial membranes. AB - Activities of testosterone, nifedipine, and midazolam oxidation by recombinant cytochrome P-450 (P-450) 3A4 coexpressed with human NADPH-P-450 reductase (NPR) in bacterial membranes (CYP3A4/NPR membranes) were determined in comparison with those of other recombinant systems and of human liver microsomes with high contents of CYP3A4. Growth conditions for Escherichia coli transformed with the bicistronic construct affected expression levels of CYP3A4 and NPR; an excess of NPR over P-450 in membrane preparations enhanced CYP3A4-dependent testosterone 6beta-hydroxylation activities of the CYP3A4/NPR membranes. Cytochrome b(5) (b(5)) and apolipoprotein b(5) further enhanced the testosterone 6beta hydroxylation activities of CYP3A4/NPR membranes after addition to either bacterial membranes or purified enzymes. NPR was observed to enhance catalytic activity when added to the CYP3A4/NPR membranes, either in the form of bacterial membranes or as purified NPR (in combination with cholate and b(5)). Apparent maximal activities of testosterone 6beta-hydroxylation in CYP3A4/NPR membranes were obtained when the molar ratio of CYP3A4/NPR/b(5) was adjusted to 1:2:1 by mixing membranes containing each protein. Testosterone 6beta-hydroxylation, nifedipine oxidation, and midazolam 4- and 1'-hydroxylation activities in CYP3A4/NPR membranes plus b(5) systems were similar to those measured with microsomes of insect cells coexpressing CYP3A4 with NPR and/or of human liver microsomes, based on equivalent CYP3A4 contents. These results suggest that CYP3A4/NPR membrane systems containing b(5) are very useful models for prediction of the rates for liver microsomal CYP3A4-dependent drug oxidations. PMID- 10460799 TI - The reductive metabolism of nitric oxide in hepatocytes: possible interaction with thiols. AB - Nitric oxide (NO) is both an endogenously generated species and the active species released from a variety of important drugs. Due to its endogenous generation and use as a therapeutic agent, the metabolism and fate of NO is of interest and concern. To date, most attention regarding the metabolism and fate of NO has been paid to its oxidized metabolites. Due to the reducing environment of cells, we considered that NO may also undergo reductive metabolism as well. Therefore, we have examined the reductive metabolism of NO by hepatocytes. Generation of nitrous oxide (N(2)O) was used as an indication of NO reduction. Indeed, we observed that NO could be reduced to N(2)O by the cytosolic fraction of hepatocytes. The N(2)O production was partially inhibited by the thiol modifying agent, N-ethylmaleimide and thiol consumption was observed during N(2)O formation. Thus, our results indicate that NO reduction is feasible and likely occurs via a thiol-dependent process. PMID- 10460800 TI - Glucuronidation of the lung carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanol (NNAL) by rat UDP-glucuronosyltransferase 2B1. AB - 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and its major metabolite, 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), are potent lung carcinogens in animals. UDP-glucuronosyltransferase (UGT)-mediated glucuronidation of NNAL is a potentially important detoxification pathway for these carcinogens. To identify the UGT isozyme(s) involved in this pathway, we examined the glucuronidation of NNAL in rat liver microsomes and homogenates from cell lines overexpressing specific UGT isozymes. NNAL glucuronidation was induced in liver microsomes from rats treated with family 2 UGT inducers including phenobarbitol and 3, 5-di-tert butyl-4-hydroxytoluene, which exhibited 1.7- and 2.6-fold higher rates of glucuronidation than microsomes from control rats. The rates of NNAL glucuronidation in liver microsomes from GUNN (deficient in family 1 UGTs) and RHA parental control rats were similar. All rat liver microsomes used in the present study catalyzed the glucuronidation of (S)-NNAL at a rate between 3.5 and 5.5 times that of the glucuronidation of (R)-NNAL. Liver microsomes from Wistar rats exhibiting the low-androsterone glucuronidation phenotype characteristic of the UGT2B2-deficient genotype glucuronidated NNAL at a rate similar to microsomes from Wistar rats exhibiting the high-androsterone glucuronidation phenotype/UGT2B2 [+] genotype. Homogenates from UGT2B1-overexpressing cells catalyzed the glucuronidation of NNAL at a K(m) of 745 microM. As with rat liver microsomes, NNAL-Gluc I was the major diastereomer formed by UGT2B1. Glucuronidation of NNAL was not detected with homogenates from UGT2B12 overexpressing cells. These results suggest that UGT2B1 plays an important role in the glucuronidation of NNAL in the rat. PMID- 10460801 TI - Rapid characterization of the major drug-metabolizing human hepatic cytochrome P 450 enzymes expressed in Escherichia coli. AB - The major drug-metabolizing human hepatic cytochrome P-450s (CYPs; CYP1A2, 2C9, 2C19, 2D6, and 3A4) coexpressed functionally in Escherichia coli with human NADPH P-450 reductase have been validated as surrogates to their counterparts in human liver microsomes (HLM) using automated technology. The dealkylation of ethoxyresorufin, dextromethorphan, and erythromycin were all shown to be specific reactions for CYP1A2, CYP2D6, and CYP3A4 that allowed direct comparison with kinetic data for HLM. For CYP2C9 and CYP2C19, the kinetics for the discrete oxidations of naproxen and diazepam were compared to data obtained using established, commercial CYP preparations. Turnover numbers of CYPs expressed in E. coli toward these substrates were generally equal to or even greater than those of the major commercial suppliers [CYP1A2 (ethoxyresorufin), E. coli 0.6 +/ 0.2 min(-1) versus B lymphoblasts 0.4 +/- 0.1 min(-1); CYP2C9 (naproxen), 6.7 +/ 0.9 versus 4.9 min(-1); CYP2C19 (diazepam), 3.7 +/- 0.3 versus 0.2 +/- 0.1 min( 1); CYP2D6 (dextromethorphan), 4.7 +/- 0.1 versus 4.4 +/- 0.1 min(-1); CYP3A4 (erythromycin), 3 +/- 1.2 versus 1.6 min(-1)]. The apparent K(m) values for the specific reactions were also similar (K(m) ranges for expressed CYPs and HLM were: ethoxyresorufin 0.5-1.0 microM, dextromethorphan 1.3-5.9 microM, and erythromycin 18-57 microM), indicating little if any effect of N-terminal modification on the E. coli-expressed CYPs. The data generated for all the probe substrates by HLM and recombinant CYPs also agreed well with literature values. In summary, E. coli-expressed CYPs appear faithful surrogates for the native (HLM) enzyme, and these data suggest that such recombinant enzymes may be suitable for predictive human metabolism studies. PMID- 10460802 TI - The role of CYP2C19 in the metabolism of (+/-) bufuralol, the prototypic substrate of CYP2D6. AB - Upon characterization of baculovirus-expressed cytochrome P-450 (CYP) 2C19, it was observed that this enzyme metabolized (+/-) bufuralol to 1'hydroxybufuralol, a reaction previously understood to be selectively catalyzed by CYP2D6. The apparent K(m) for this reaction was 36 microM with recombinant CYP2C19, approximately 7-fold higher than for recombinant CYP2D6. The intrinsic clearance for this reaction was 37-fold higher with CYP2D6 than for CYP2C19. The involvement of human CYP1A2 in bufuralol 1'-hydroxylation was also confirmed using the recombinant enzyme. Using S-mephenytoin as an inhibitor, the K(i) for inhibition of recombinant CYP2C19-mediated bufuralol hydroxylation was 42 microM, which is the approximate K(m) for recombinant CYP2C19-mediated S-mephenytoin metabolism. The classic CYP2D6 inhibitors quinidine and quinine showed no inhibition of CYP2C19-catalyzed bufuralol metabolism at concentrations that abolished CYP2D6-mediated bufuralol metabolism. Ticlopidine, a potent inhibitor of CYP2C19 and CYP2D6, inhibited bufuralol 1'-hydroxylation by each of these enzymes equipotently. In human liver microsomes that are known to be deficient in CYP2D6 activity, it was shown that in the presence of quinidine, the K(m) shifted from 14 to 38 microM. This is consistent with the K(m) determination for recombinant CYP2C19 of 36 microM. In human liver microsomes that have high CYP2D6 and CYP2C19 activity, the K(m) shifted to 145 microM in the presence of S mephenytoin and quinidine, consistent with the K(m) determined for CYP1A2. This data suggests that bufuralol, and possibly other CYP2D6 substrates, have the potential to be metabolized by CYP2C19. PMID- 10460803 TI - Multiple cytochrome P-450s involved in the metabolism of terbinafine suggest a limited potential for drug-drug interactions. AB - Biotransformation pathways and the potential for drug-drug interactions of the orally active antifungal terbinafine were characterized using human liver microsomes and recombinant human cytochrome P-450s (CYPs). The terbinafine metabolites represented four major pathways: 1) N-demethylation, 2) deamination, 3) alkyl side chain oxidation, and 4) dihydrodiol formation. Michaelis-Menten kinetics for the pathways revealed mean K(m) values ranging from 4.4 to 27.8 microM, and V(max) values of 9.8 to 82 nmol/h/mg protein. At least seven CYP enzymes are involved in terbinafine metabolism. Recombinant human CYPs predict that CYP2C9, CYP1A2, and CYP3A4 are the most important for total metabolism. N demethylation is primarily mediated by CYP2C9, CYP2C8, and CYP1A2; dihydrodiol formation by CYP2C9 and CYP1A2; deamination by CYP3A4; and side chain oxidation equally by CYP1A2, CYP2C8, CYP2C9, and CYP2C19. Additionally, characteristic CYP substrates inhibited pathways of terbinafine metabolite formation, confirming the involvement of multiple enzymes. The deamination pathway was mainly inhibited by CYP3A inhibitors, including troleandomycin and azole antifungals. Dihydrodiol formation was inhibited by the CYP1A2 inhibitor furafylline. Terbinafine had little or no effect on the metabolism of many characteristic CYP substrates. Terbinafine, however, is a competitive inhibitor of the CYP2D6 reaction, dextromethorphan O-demethylation (K(i) = 0.03 microM). In summary, terbinafine is metabolized by at least seven CYPs. The potential for terbinafine interaction with other drugs is predicted to be insignificant with the exception that it may inhibit the metabolism of CYP2D6 substrates. Clinical trials are needed to assess the relevance of these findings. PMID- 10460804 TI - Oxidative metabolism of monensin in rat liver microsomes and interactions with tiamulin and other chemotherapeutic agents: evidence for the involvement of cytochrome P-450 3A subfamily. AB - Monensin (MON) is an ionophore antibiotic widely used in veterinary practice as a coccidiostatic or a growth promoter. The aims of this study were to characterize the P-450 isoenzyme(s) involved in the biotransformation of the ionophore and to investigate how this process may be affected by tiamulin and other chemotherapeutic agents known to produce toxic interactions with MON when administered concurrently in vivo. In liver microsomes from untreated rats (UT) or from rats pretreated, respectively, with ethanol (ETOH), beta-naphthoflavone (betaNAF), phenobarbital (PB), pregnenolone 16alpha-carbonitrile (PCN), or dexamethasone (DEX), the rate of MON O-demethylation was the following: DEX > PCN > PB >> UT = ETOH > betaNAF; similar results were obtained by measuring total MON metabolism. In addition, the extent of triacetyloleandomycin-mediated P-450 complexes was greatly reduced by the prior addition of 100 microM MON. In DEX treated microsomes, MON O-demethylation was found to fit monophasic Michaelis Menten kinetics (K(M) = 67.6 +/- 0.01 microM; V(max) = 4.75 +/- 0.76 nmol/min/mg protein). Tiamulin markedly inhibited this activity in an apparent competitive manner, with a calculated K(i) (Dixon plot) of 8.2 microM and an IC(50) of about 25 microM. At the latter concentration, only ketoconazole or metyrapone, which can bind P-450 3A, inhibited MON O-demethylase to a greater extent than tiamulin, whereas alpha-naphthoflavone, chloramphenicol, or sulphametasine was less effective. These results suggest that P-450 3A plays an important role in the oxidative metabolism of MON and that compounds capable of binding or inhibiting this isoenzyme could be expected to give rise to toxic interactions with the ionophore. PMID- 10460805 TI - Liquid chromatography/mass spectrometry and high-field nuclear magnetic resonance characterization of novel mixed diconjugates of the non-nucleoside human immunodeficiency virus-1 reverse transcriptase inhibitor, efavirenz. AB - Efavirenz (Sustiva) is a potent and specific inhibitor of the HIV-1 reverse transcriptase and is approved for the treatment of HIV infection. The metabolism of efavirenz in different species has been described previously. Efavirenz is primarily metabolized in rats to the glucuronide conjugate of 8-OH efavirenz. Electrospray ionization-liquid chromatography/mass spectrometry analyses of bile samples from rats dosed with either efavirenz or with 8-OH efavirenz revealed three polar metabolites, M9, M12, and M13, with pseudomolecular ions [M-H](-) at m/z 733, 602, and 749, respectively. The characteristic mass spectral fragmentation patterns obtained for metabolites M9 and M13 suggested that these were glutathione-sulfate diconjugates, and the presence of a glutathione moiety in metabolite M9 was confirmed by liquid chromatograpy/nuclear magnetic resonance (NMR) analysis of bile extracts. Metabolite M12 was characterized by liquid chromatography/mass spectrometry as a glucuronide-sulfate diconjugate. Unambiguous structures of M9, M12, and M13 were obtained from one-dimensional proton and carbon NMR as well as proton-proton (correlated spectroscopy, two dimensional shift correlation), proton-carbon heteronuclear multiple quantum correlation, and long-range proton-carbon (heteronuclear multiple bond correlation) correlated two-dimensional NMR analyses of metabolites isolated from rat bile. The mass spectral and NMR analyses of M10, which was isolated from rat urine, suggested a cysteinylglycine-sulfate diconjugate. The isolation of these polar metabolites for further characterization by NMR was aided by mass spectral analyses of HPLC fractions and solid phase extraction extracts during the isolation steps. The complete characterization of these novel diconjugates demonstrates that further phase II metabolism of polar conjugates such as sulfates could take place in vivo. PMID- 10460806 TI - Studies on sulfation of synthesized metabolites from the local anesthetics ropivacaine and lidocaine using human cloned sulfotransferases. AB - The metabolism of the local anesthetics lidocaine and ropivacaine (ropi) involves several steps in humans. Lidocaine is mainly hydrolyzed and hydroxylated to 4-OH 2,6-xylidine (4-OH-xyl). The metabolism of ropi, involving dealkylation and hydroxylation, gives rise to 3-OH-ropi, 4-OH-ropi, 3-OH-2'6'-pipecoloxylidide (3 OH-PPX), and 2-OH-methyl-ropi. Because the metabolites are hydroxylated, they are particularly prone to subsequent Phase II conjugation reactions such as sulfation and glucuronidation. This study focused on the in vitro sulfation of these metabolites as well as another suspected metabolite of ropi, 2-carboxyl-ropi. All the metabolites were synthesized for the subsequent enzymatic studies. Five cloned human sulfotransferases (STs) were used in this study, namely, the phenol sulfating form of ST (P-PST-1), the monoamine-sulfating form of ST (M-PST), estrogen-ST (EST), ST1B2, and dehydroepiandrosterone-ST (DHEA-ST), all of which are expressed in human liver. The results demonstrate that all of the metabolites except 2-OH-methyl-ropi and 2-carboxyl-ropi can be sulfated. It was also found that all of the STs can conjugate the remaining hydroxylated metabolites except DHEA-ST. However, there are large differences in the capacity of the individual human ST isoforms to conjugate the different metabolites. P-PST-1 sulfates 3-OH PPX, 3-OH-ropi, and 4-OH-xyl; M-PST and EST conjugate 3-OH-PPX, 3-OH-ropi, and 4 OH-ropi whereas ST1B2 sulfates only 4-OH-xyl. The most extensively sulfated ropi metabolite is 3-OH-PPX. In conclusion, all of the hydroxylated metabolites of lidocaine and ropi can be sulfated if the hydroxyl group is attached to the aromatic ring in the metabolites. The human ST enzymes that are considered to be responsible for the sulfation of these metabolites in vivo are P-PST-1, M-PST, EST, and ST1B2. These enzymes are also found in the liver; this is the most important tissue for the metabolism of ropi in humans, demonstrated by. PMID- 10460807 TI - Human and Escherichia coli beta-glucuronidase hydrolysis of glucuronide conjugates of benzidine and 4-aminobiphenyl, and their hydroxy metabolites. AB - Individuals exposed to carcinogenic aromatic amines excrete arylamine N- and O glucuronide metabolites. This study assessed the susceptibility of selected glucuronides to hydrolysis by human and Escherichia coli beta-glucuronidase. N- or O-glucuronides were prepared with the following aglycones: benzidine, N acetylbenzidine, N'-hydroxy-N-acetylbenzidine, N-hydroxy-N-acetylbenzidine, N hydroxy-N,N'-diacetylbenzidine, 3-hydroxy-N,N'-diacetylbenzidine, 3-hydroxy benzidine, 4-aminobiphenyl, N-hydroxy-4-aminobiphenyl, and N-hydroxy-N-acetyl-4 aminobiphenyl. The (3)H- and (14)C-labeled glucuronides were prepared with human or rat liver microsomes using UDP-glucuronic acid as cosubstrate. Each of the 10 glucuronides (6-12 microM) was incubated at pH 5.5 or 7.0 with either human recombinant (pure) or E. coli (commercial preparation) beta-glucuronidase for 30 min at 37 degrees C. Hydrolysis was measured by HPLC. Reaction conditions were optimized, using the O-glucuronide of N-hydroxy-N,N'-diacetylbenzidine. Both enzymes preferentially hydrolyzed O-glucuronides over N-glucuronides and distinguished between structural isomers. With E. coli beta-glucuronidase at pH 7.0, selectivity was demonstrated by the complete hydrolysis of N-hydroxy-N acetyl-4-aminobiphenyl O-glucuronide in the presence of N-acetylbenzidine N glucuronide, which was not hydrolyzed. Metabolism by both enzymes was completely inhibited by the specific beta-glucuronidase inhibitor saccharic acid-1,4-lactone (0.5 mM). The concentration of human beta-glucuronidase necessary to achieve significant hydrolysis of glucuronides was substantially more than the amount of enzyme reported previously to be present in urine under either normal or pathological conditions. The bacterial enzyme may hydrolyze O-glucuronides, but not N-glucuronides, in urine at neutral pH. Thus, the nonenzymatic hydrolysis of N-glucuronides by acidic urine is likely a more important source of free amine than enzymatic hydrolysis. PMID- 10460808 TI - Cytochrome P-450 3A4 and 2C8 are involved in zopiclone metabolism. AB - Zopiclone is a widely prescribed, nonbenzodiazepine hypnotic that is extensively metabolized by the liver in humans. The aim of the present study was to identify the human cytochrome P-450 (CYP) isoforms involved in zopiclone metabolism in vitro. Zopiclone metabolism was studied with different human liver microsomes and a panel of heterologously expressed human CYPs (CYP1A2, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, and 3A4). In human liver microsomes, zopiclone was metabolized into N desmethyl-zopiclone (ND-Z) and N-oxide-zopiclone (NO-Z) with the following K(m) and V(m) of 78 +/- 5 and 84 +/- 19 microM, 45 +/- 1 and 54 +/- 5 pmol/min/mg for ND-Z and NO-Z generation, respectively. Ketoconazole (CYP3A inhibitor) inhibited approximately 40% of the generation of both metabolites, sulfaphenazole (CYP2C inhibitor) inhibited the formation of ND-Z, whereas alpha-naphtoflavone (CYP1A), quinidine (CYP2D6), and chlorzoxazone (CYP2E1) did not affect zopiclone metabolism. The generation of ND-Z and NO-Z were highly correlated to testosterone 6beta-hydroxylation (CYP3A activity, r = 0.95 and 0.92, respectively; p =.0001), and ND-Z was highly correlated to CYP2C8 activity (paclitaxel 6alpha-hydroxylase; r = 0.76, p =.004). Recombinant CYP2C8 had the highest enzymatic activity toward zopiclone metabolism into both its metabolites, followed by CYP2C9 and 3A4. CYP3A4 is the major enzyme involved in zopiclone metabolism in vitro, and CYP2C8 contributes significantly to ND-Z formation. PMID- 10460809 TI - Effect of the acute-phase response on the pharmacokinetics of chlorzoxazone and cytochrome P-450 2E1 in vitro activity in rats. AB - The acute-phase response is known to produce alterations in hepatic cytochrome P 450 (CYP) expression. Lipopolysaccharide (LPS), a well known inducer of acute phase response decreases hepatic CYP2E1 in vitro activity in rats. This study was designed to determine if LPS administration produced alterations in the pharmacokinetics of chlorzoxazone (CZN), a marker for CYP2E1 expression. Sprague Dawley rats were administered a single i.p. injection of LPS (5 mg/kg) or saline control approximately 24 h before a single i.v. bolus dose of CZN (15 mg/kg). Serial blood samples were collected over a 120-min period to quantitate CZN plasma concentrations and protein binding. In addition, livers were removed and processed for evaluating in vitro CYP2E1 protein concentrations and activity. Systemic clearance decreased by 35% in LPS-treated rats, whereas half-life and steady-state volume of distribution increased by 167 and 66%, respectively. The plasma free-fraction of CZN increased 2-fold after LPS treatment. The CZN intrinsic clearance decreased in LPS rats by 71% compared with control values. The CYP2E1 liver microsomal activity decreased between 55 and 75% along with a 41% decrease in CYP2E1 protein concentration. The CZN intrinsic clearance was significantly correlated with both the CZN and p-nitrophenol liver microsomal activity (r = 0.97 and r = 0.91, respectively). This study demonstrated that LPS administration produced expected reductions in the in vivo intrinsic clearance of CZN, and these changes were highly correlated with in vitro activity studies. In addition, LPS produced significant increases in the steady-state volume of distribution of CZN secondary to reductions in its plasma protein binding. PMID- 10460810 TI - Effect of antipsychotic drugs on human liver cytochrome P-450 (CYP) isoforms in vitro: preferential inhibition of CYP2D6. AB - The ability of antipsychotic drugs to inhibit the catalytic activity of five cytochrome P-450 (CYP) isoforms was compared using in vitro human liver microsomal preparations to evaluate the relative potential of these drugs to inhibit drug metabolism. The apparent kinetic parameters for enzyme inhibition were determined by nonlinear regression analysis of the data. All antipsychotic drugs tested competitively inhibited dextromethorphan O-demethylation, a selective marker for CYP2D6, in a concentration-dependent manner. Thioridazine and perphenazine were the most potent, with IC(50) values (2.7 and 1.5 microM) that were comparable to that of quinidine (0.52 microM). The estimated K(i) values for CYP2D6-catalyzing dextrorphan formation were ranked in the following order: perphenazine (0.8 microM), thioridazine (1.4 microM), chlorpromazine (6.4 microM), haloperidol (7.2 microM), fluphenazine (9.4 microM), risperidone (21.9 microM), clozapine (39.0 microM), and cis-thiothixene (65.0 microM). No remarkable inhibition of other CYP isoforms was observed except for moderate inhibition of CYP1A2-catalyzed phenacetin O-deethylation by fluphenazine (K(i) = 40.2 microM) and perphenazine (K(i) = 65.1). The estimated K(i) values for the inhibition of CYP2C9, 2C19, and 3A were >300 microM in almost all antipsychotics tested. These results suggest that antipsychotic drugs exhibit a striking selectivity for CYP2D6 compared with other CYP isoforms. This may reflect a remarkable commonality of structure between the therapeutic targets for these drugs, the transporters, and metabolic enzymes that distribute and eliminate them. Clinically, coadministration of these medicines with drugs that are primarily metabolized by CYP2D6 may result in significant drug interactions. PMID- 10460811 TI - Absence of host-site influence on angiogenesis, blood flow, and permeability in transplanted RG-2 gliomas. AB - The host site is believed to regulate tumor angiogenesis, which could result in site-dependent drug delivery parameters, greatly affecting experimental tumor research. In RG-2 rat gliomas we measured cellular proliferation; cell cycle time was the same for RG-2 cells in brain and s.c. tumors (25 h), and was the same for endothelial cells in these tumors (46 h). We measured the transcapillary transfer constant (K) of alpha-aminoisobutyric acid and blood flow (F) with iodoantipyrine in RG-2 gliomas transplanted into brain, liver, kidney, muscle, s.c. tissue, and into the abdominal cavity. Data was evaluated by quantitative autoradiography and direct tissue sampling. The variation of F (12.6-84.0 ml/g/min) and K (26.1-49.2 microl/g/min) in RG-2 tumors in the different host sites was less than in surrounding tumor-free tissue (F = 20-1500 ml/g/min and K = 1.6-700 microl/g/min). In contrast to other models, RG-2 does not result in tumors with host site-dependent behavior. The RG-2 tumor cells appear to participate in, if not dominate, the angiogenesis process regardless of the host site. Values of F and K were more dependent on tumor topography (center versus periphery) and local histological features (necrosis versus viable tumor) than host site. We believe that the methods used for data acquisition may introduce as much variability in Results as the tumors themselves and that to better understand how tumor angiogenesis affects the vascular phenotype, comparative studies are needed to validate the results obtained with newer methodologies. PMID- 10460812 TI - Diurnal variation in the metabolism of S-carboxymethyl-L-cysteine in humans. AB - The routes of metabolism of S-carboxymethyl-L-cysteine in humans are dependent on the time of dosing. Administration of 750 mg of S-carboxymethyl-L-cysteine (Day 1) during the day at 8:00 AM followed by a 8:00 AM to 4:00 PM urine collection revealed that S-carboxymethyl-L-cysteine S-oxide was the major urinary metabolite produced. The 4:00 PM to midnight urine collection resulted in S (carboxymethylthio)-L-cysteine being identified as the major urinary metabolite. However, the administration of 750 mg of S-carboxymethyl-L-cysteine (day 15) during the night at midnight and analysis of the midnight to 8:00 AM urine collection found that thiodiglycolic acid was the major urinary metabolite, whereas thiodiglycolic S-oxide was identified as the major urinary metabolite in the 8:00 AM to 4:00 PM urine collection. A diurnal variation in the metabolism of S-carboxymethyl-L-cysteine was seen and, in particular, the timing of S carboxymethyl-L-cysteine administration had a profound effect on the identity of urinary S-oxide metabolites produced. After administration at 8:00 AM the urinary S-oxides produced were S-carboxymethyl-L-cysteine S-oxide and S-methyl-L-cysteine S-oxide but at midnight the major urinary S-oxide metabolite produced was thiodiglycolic acid S-oxide. PMID- 10460813 TI - Early revascularization in acute myocardial infarction complicated by cardiogenic shock. SHOCK Investigators. Should We Emergently Revascularize Occluded Coronaries for Cardiogenic Shock. AB - BACKGROUND: The leading cause of death in patients hospitalized for acute myocardial infarction is cardiogenic shock. We conducted a randomized trial to evaluate early revascularization in patients with cardiogenic shock. METHODS: Patients with shock due to left ventricular failure complicating myocardial infarction were randomly assigned to emergency revascularization (152 patients) or initial medical stabilization (150 patients). Revascularization was accomplished by either coronary-artery bypass grafting or angioplasty. Intraaortic balloon counterpulsation was performed in 86 percent of the patients in both groups. The primary end point was mortality from all causes at 30 days. Six-month survival was a secondary end point. RESULTS: The mean age of the patients was 66+/-10 years, 32 percent were women and 55 percent were transferred from other hospitals. The median time to the onset of shock was 5.6 hours after infarction, and most infarcts were anterior in location. Ninety-seven percent of the patients assigned to revascularization underwent early coronary angiography, and 87 percent underwent revascularization; only 2.7 percent of the patients assigned to medical therapy crossed over to early revascularization without clinical indication. Overall mortality at 30 days did not differ significantly between the revascularization and medical-therapy groups (46.7 percent and 56.0 percent, respectively; difference, -9.3 percent; 95 percent confidence interval for the difference, -20.5 to 1.9 percent; P=0.11). Six-month mortality was lower in the revascularization group than in the medical-therapy group (50.3 percent vs. 63.1 percent, P=0.027). CONCLUSIONS: In patients with cardiogenic shock, emergency revascularization did not significantly reduce overall mortality at 30 days. However, after six months there was a significant survival benefit. Early revascularization should be strongly considered for patients with acute myocardial infarction complicated by cardiogenic shock. PMID- 10460814 TI - Surgery to cure the Zollinger-Ellison syndrome. AB - BACKGROUND AND METHODS: The role of surgery in patients with the Zollinger Ellison syndrome is controversial. To determine the efficacy of surgery in patients with this syndrome, we followed 151 consecutive patients who underwent laparotomy between 1981 and 1998. Of these patients, 123 had sporadic gastrinomas and 28 had multiple endocrine neoplasia type 1 with an imaged tumor of at least 3 cm in diameter. Tumor-localization studies and functional localization studies were performed routinely. All patients underwent surgery according to a similar operative protocol, and all patients who had surgery after 1986 underwent duodenotomy. RESULTS: The 151 patients underwent 180 exploratory operations. The mean (+/-SD) follow-up after the first operation was 8+/-4 years. Gastrinomas were found in 141 of the patients (93 percent), including all of the last 81 patients to undergo surgery. The tumors were located in the duodenum in 74 patients (49 percent) and in the pancreas in 36 patients (24 percent); however, primary tumors were found in lymph nodes in 17 patients (11 percent) and in another location in 13 patients (9 percent). The primary location was unknown in 24 patients (16 percent). Among the patients with sporadic gastrinomas, 34 percent were free of disease at 10 years, as compared with none of the patients with multiple endocrine neoplasia type 1. The overall 10-year survival rate was 94 percent. CONCLUSIONS: All patients with the Zollinger-Ellison syndrome who do not have multiple endocrine neoplasia type 1 or metastatic disease should be offered surgical exploration for possible cure. PMID- 10460815 TI - Obstetrical outcomes among women with extrapulmonary tuberculosis. AB - BACKGROUND: The prevalence of tuberculosis, especially extrapulmonary tuberculosis, is increasing worldwide. Because information on the outcome of pregnancy among women with extrapulmonary tuberculosis is limited, we studied the course of pregnancy and labor and the perinatal outcome in these women and their infants. METHODS: From 1983 to 1993, we followed 33 pregnant women who had extrapulmonary tuberculosis (12 with tuberculous lymphadenitis and 9 with intestinal, 7 with skeletal, 2 with renal, 2 with meningeal, and 1 with endometrial tuberculosis) through their deliveries. Of the 33, 29 received antituberculosis treatment during pregnancy. The antenatal complications, intrapartum events, and perinatal outcomes were compared with those among 132 healthy pregnant women without tuberculosis who were matched for age, parity, and socioeconomic status. RESULTS: Tuberculous lymphadenitis did not affect the course of pregnancy or labor or the perinatal outcome. However, as compared with the control women, the 21 women with tubercular involvement of other extrapulmonary sites had higher rates of antenatal hospitalization (24 percent vs. 2 percent, P< 0.001), infants with low Apgar scores (< or =6) soon after birth (19 percent vs. 3 percent, P=0.01), and low-birth-weight (<2500 g) infants (33 percent vs. 11 percent, P=0.01). CONCLUSIONS: Extrapulmonary tuberculosis that is confined to the lymph nodes has no effect on obstetrical outcomes, but tuberculosis at other extrapulmonary sites does adversely affect the outcome of pregnancy. PMID- 10460816 TI - A prospective study of walking as compared with vigorous exercise in the prevention of coronary heart disease in women. AB - BACKGROUND: The role of walking, as compared with vigorous exercise, in the prevention of coronary heart disease remains controversial, and data for women on this topic are sparse. METHODS: We prospectively examined the associations between the score for total physical activity, walking, and vigorous exercise and the incidence of coronary events among 72,488 female nurses who were 40 to 65 years old in 1986. Participants were free of diagnosed cardiovascular disease or cancer at the time of entry and completed serial detailed questionnaires about physical activity. During eight years of follow-up, we documented 645 incident coronary events (nonfatal myocardial infarction or death from coronary disease). RESULTS: There was a strong, graded inverse association between physical activity and the risk of coronary events. As compared with women in the lowest quintile group for energy expenditure (expressed as the metabolic-equivalent [MET] score), women in increasing quintile groups had age-adjusted relative risks of 0.77, 0.65, 0.54, and 0.46 for coronary events (P for trend <0.001). In multivariate analyses, the inverse gradient remained strong (relative risks, 0.88, 0.81, 0.74, and 0.66 for women in increasing quintile groups as compared with those in the lowest quintile group; P for trend=0.002). Walking was inversely associated with the risk of coronary events; women in the highest quintile group for walking, who walked the equivalent of three or more hours per week at a brisk pace, had a multivariate relative risk of 0.65 (95 percent confidence interval, 0.47 to 0.91) as compared with women who walked infrequently. Regular vigorous exercise (> or =6 MET) was associated with similar risk reductions (30 to 40 percent). Sedentary women who became active in middle adulthood or later had a lower risk of coronary events than their counterparts who remained sedentary. CONCLUSIONS: These prospective data indicate that brisk walking and vigorous exercise are associated with substantial and similar reductions in the incidence of coronary events among women. PMID- 10460817 TI - Images in clinical medicine. Mesenteric cyst. PMID- 10460819 TI - Treatment of diabetic retinopathy. PMID- 10460818 TI - The control of labor. PMID- 10460820 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 26-1999. A three-week-old girl with pyloric stenosis and an unexpected operative finding. PMID- 10460821 TI - Goodbye, for now. PMID- 10460822 TI - Early revascularization in cardiogenic shock--a positive view of a negative trial. PMID- 10460823 TI - Surgery for the Zollinger-Ellison syndrome. PMID- 10460825 TI - Correction: Concurrent Cisplatin-Based Radiotherapy and Chemotherapy for Locally Advanced Cervical Cancer. PMID- 10460824 TI - How to resolve an ethical dilemma concerning randomized clinical trials. PMID- 10460826 TI - Enhancing therapeutic glycoprotein production in Chinese hamster ovary cells by metabolic engineering endogenous gene control with antisense DNA and gene targeting. AB - Recombinant glycoprotein therapeutics have proven to be invaluable pharmaceuticals for the treatment of chronic and life-threatening diseases. Although these molecules are extraordinarily efficacious, many diseases have high dosage requirements of several hundred milligrams of protein for each administration. Multiple doses at this level are often required for treatment. One of the major challenges currently facing the biotechnology industry is the development of large-scale, cost-effective production and manufacturing processes of these biologically synthesized molecules. Metabolic engineering of animal cell expression hosts promises to address this challenge by substantially enhancing recombinant protein quality, productivity, and biological activity. In this report, we describe a novel approach to metabolic engineering in Chinese hamster ovary cells by control of endogenous gene expression. Analysis of the advantages and limitations of using antisense DNA and gene targeting as a means of control are discussed and several gene candidates for regulation with these techniques are identified. Practical considerations for using these technologies to reduce the levels of the CHO cell sialidase (Warner et al., Glycobiology, 3, 455-463, 1993) as a model gene system for regulation are also presented. PMID- 10460827 TI - Molecular cloning, expression and exon/intron organization of the bovine beta galactoside alpha2,6-sialyltransferase gene. AB - In this study, we report the first isolation and characterization of a bovine sialyltransferase gene. Bovine cDNAs prepared from different tissues contain an open-reading frame encoding a 405 amino acid sequence showing 83%, 75%, and 60% identity with human, murine, and chicken ST6Gal I (beta-galactoside alpha2,6 sialyltransferase) sequences, respectively. When transfected into COS-7 cells, a recombinant enzyme was obtained which catalyzed the in vitro alpha2, 6 sialylation of LacNAc (NeuAcalpha2-6Galbeta1-4GlcNAc) and LacdiNAc (NeuAcalpha2 6GalNAcbeta1-4GlcNAc) acceptor substrates. The K (m) values were 2.8 and 6.9 mM, respectively. Different relative efficiencies (Vmax/Km) for the two precursors (36 for LacNAc and 4.3 for LacdiNAc) were observed. Bovine ST6Gal I gene consists of four 5'-untranslated exons E(-2) to E(1), and five coding exons from E(2) to E(6). This later carries a 3'-untranslated region of 2. 7 kb. Gene sequence spans at least 80 kb of genomic DNA. Two processed pseudogenes have been identified. They are 94.3 and 95.6% similar to the bovine cDNA, respectively. Three families of mRNA isoforms were isolated. They differed by their 5'-untranslated regions and could be generated by three tissue-specific promoters. Family 1 is made up of exons E(-2) and E(1) to E(6), family 2 of exons E(-1) to E(6), and family 3 of exons E(1) to E(6). Tissular distribution of transcript families appears noticeably different than those described in human and rat. PMID- 10460828 TI - Naturally occurring anti-alpha-galactosyl antibodies: relationship to xenoreactive anti-alpha-galactosyl antibodies. AB - Antibodies produced by an individual without a known history of sensitization to the relevant antigen are called "natural" antibodies. Some natural antibodies, called xenoreactive antibodies, react with the cells of foreign species. Most xenoreactive antibodies in humans and higher primates bind to a nonreducing terminal galactose expressed by pigs and other lower mammals. Although human natural antibodies which bind to one or more of a variety of terminal alpha galactosyl structures have been identified previously, the antigen recognized by anti-alpha-galactosyl antibodies on the cells of foreign species is thought to be exclusively Galalpha1-3Gal. Thus, anti-alpha-galactosyl antibodies which do not react with Galalpha1-3Gal are thought to be nonxenoreactive. Here, we identify natural antibodies in human serum which bind to Galalpha1-6Hexosepyrranosides but not Galalpha1-3Gal, indicating that these antibodies are not xenoreactive. Various lower mammals were found to have natural anti-Galalpha1-2Gal antibodies in their sera, suggesting that at least some anti-Galalpha1-2Gal antibodies might not be xenoreactive and indicating, surprisingly, that anti-alpha-galactosyl antibodies are much more phylogenetically disperse than previously known. Also surprising was the finding that some natural antibodies which bind to Galalpha1 3Gal in vitro do not bind to porcine xenografts. These studies show that naturally occurring anti-alpha-galactosyl antibodies in mammalian serum include antibodies with a greater variety of reactivities than previously thought, only some of which would bind to a porcine xenograft. Further, these studies show that the methods used to detect anti-alpha-galactosyl antibodies of relevance in xenotransplantation must be carefully evaluated to avoid detection of anti-alpha galactosyl antibodies which would not bind to a porcine organ and which therefore are not involved in xenograft rejection. PMID- 10460829 TI - Forssman penta- and tetraglycosylceramide are xenoantigens of ostrich kidney and liver. AB - The heterophile antigens Galalpha1-->3Gal and N-glycolylneuraminic acid are the major obstacle to grafting mammal organs, especially from pig, to man. Lack of expression of these common xenoantigens by birds has raised interest in ostrich as a potential organ donor for xenotransplantation. Glycosphingolipids of ostrich liver and kidney were investigated for their carbohydrate determinants. Both organs were found similar in their glycolipid composition with three major species, mono-, di-, and pentaglycosylceramide. The pentaglycosylceramide was characterized as the Forssman antigen. In both organs, the ceramide portion was highly hydroxylated with prevalence of alpha-hydroxylated fatty acids, C18 phytosphingosine in kidney and C18 sphingosine in liver Forssman glycolipid. These data indicate that hydroxylation of kidney glycosphingolipids, which is found in mammals, has been maintained since the divergence of birds from other vertebrates. Characterization of a minor glycolipid as a Forssman tetraglycosylceramide built on the galabiosylceramide core indicates that the Forssman tetraglycosylceramide also exists in vivo. Its precursors, galactosyl- and galabiosylceramide, were characterized in kidney and liver. The Forssman antigen is the third heterophile antigen against which man raises natural antibodies. Its localization in the vascular endothelium and connective tissue makes ostrich an unpromising organ or cell donor for xenotransplantation to man. PMID- 10460830 TI - Identification of two discrete peptide: N-glycanases in Oryzias latipes during embryogenesis. AB - Two different types of peptide:N-glycanase (PNGase) were identified in developing embryos of medaka fish ( Oryzias latipes ). Because the optimum pH values for their activities were acidic and neutral, they were designated as acid PNGase M and neutral PNGase M, respectively. The acid PNGase M corresponded to the enzyme that had been partially purified from medaka embryos (Seko,A., Kitajima,K., Inoue,Y. and Inoue,S. (1991) J. Biol. Chem., 266, 22110-22114). The apparent molecular weight of this enzyme was 150 K, and the optimal pH was 3.5-4.0, and the K m for L-hyosophorin was 44 microM. L-Hyosophorin is a cortical alveolus derived glycononapeptide with a large N-linked glycan chain present in the perivitelline space of the developing embryo. Acid PNGase M was competitively inhibited by a free de-N-glycosylated nonapeptide derived from L-hyosophorin. This enzyme was expressed in ovaries and embryos at all developmental stages after gastrulation, but activity was not detected in embryos at developmental stages between fertilization and gastrula. Several independent lines of evidence suggested that acid PNGase M may be responsible for the unusual accumulation of free N-glycans derived from yolk glycoproteins (Iwasaki,M., Seko,A., Kitajima,K., Inoue,Y. and Inoue,S. (1992) J. Biol. Chem., 267, 24287-24296). In contrast, the neutral PNGase M was expressed in blastoderms from the 4-8 cell stage and in cells up to early gastrula. The general significance of these findings is that they show a developmental stage-dependent expression of the two PNGase activities, and that expression of the neutral PNGase M activity occurs concomitantly with the de-N-glycosylation of L-hyosophorin. These data thus support our conclusion that the neutral PNGase M is responsible for the developmental-stage-related de-N-glycosylation of the L-hyosophorin. PMID- 10460831 TI - N-glycosylation of the carcinoembryonic antigen related cell adhesion molecule, C CAM, from rat liver: detection of oversialylated bi- and triantennary structures. AB - Rat C-CAM is a ubiquitous, transmembrane and carcinoembryonic antigen related cell adhesion molecule. The human counterpart is known as biliary glycoprotein (BGP) or CD66a. It is involved in different cellular functions ranging from intercellular adhesion, microbial receptor activity, signaling and tumor suppression. In the present study N-glycosylation of C-CAM immunopurified from rat liver was analyzed in detail. The primary sequence of rat C-CAM contains 15 potential N-glycosylation sites. The N-glycans were enzymatically released from glycopeptides, fluorescently labeled with 2-aminobenzamide, and separated by two dimensional HPLC. Oligosaccharide structures were characterized by enzymatic sequencing and MALDI-TOF-MS. Mainly bi- and triantennary complex structures were identified. The presence of type I and type II chains in the antennae of these glycans results in heterogeneous glycosylation of C-CAM. Sialylation of the sugars was found to be unusual; bi- and triantennary glycans contained three and four sialic acid residues, respectively, and this linkage seemed to be restricted to the type I chain in the antennae. Approximately 20% of the detected sugars contain these unusual numbers of sialic acids. C-CAM is the first transmembrane protein found to be oversialylated. PMID- 10460832 TI - Transcription of the beta-galactoside alpha2,6-sialyltransferase gene (SIAT1) in B-lymphocytes: cell type-specific expression correlates with presence of the divergent 5'-untranslated sequence. AB - A single gene, SIAT1, encodes ST6Gal I, the sialyltransferase that mediates transfer of alpha2,6-linked sialic acids to Galbeta1, 4GlcNAc termini of N-linked glycoproteins. In vivo, multiple SIAT1 mRNA forms, differing only in the 5' untranslated region, are expressed in a tissue-specific manner. This mRNA heterogeneity has been attributed, at least in part, to transcription from a number of physically distinct promoter regions. In mature B-lymphocytes, SIAT1 transcription initiates at P2, a regulatory region known to function only in B lineage cells. Bacterial chloramphenicol acetyltransferase (CAT) under the control of the P2 region encompassing 415 bp 5'- and 125 bp 3' of the transcriptional initiation site is efficiently expressed in Louckes, a mature B lymphoblastoid cell line. In contrast, CAT expression in Reh, a T-null/B-null precursor line, and in HepG2, a hepatoma line, are 14-fold and >25-fold less than in Louckes, respectively. The data is consistent with the presence of cis -acting regulatory elements residing both 5' and 3' of the P2 transcriptional initiation site. At least 370 bp of 5'-flanking sequence, coinciding with the inclusion of AP2 and NF-kappaB sites, is necessary for high level expression in Louckes. Exon sequences 3' of the transcription start site are also important for expression. A segment from(+)32 to(+)125 (position(+)1 is transcription start site) is capable of exerting promoter-like activity in Louckes, but not in Reh or HepG2. CAT expression by P2 is negligible in Reh cells. However, enhanced CAT activity is not accompanied by elevated mRNA levels. This observation is consistent with the relief of translational restraints imposed by the(+)32 to(+)125 region. Together, the data demonstrate that efficient and cell-specific transcription regulation in mature B lymphocytes is contained in a 495 bp P2 segment that is comprised of 370 bp of 5'-flanking region and 125 bp of transcribed region of Exon X. PMID- 10460833 TI - Characterization of human vascular endothelial cadherin glycans. AB - The glycosylation pattern of human vascular endothelial cadherin (VE-cadherin), purified from cultured human umbilical cord vein endothelial cells, was analyzed. VE-cadherin was metabolically radiolabeled with d-[6-(3)H]glucosamine, isolated by immunoprecipitation, purified by SDS-PAGE and in-gel digested with endoproteinase Asp N. Oligosaccharides were sequentially released from resulting glycopeptides and analyzed by chromatographic profiling. The results revealed that VE-cadherin carries predominantly sialylated diantennary and hybrid-type glycans in addition to some triantennary and high mannose-type species. Highly branched, tetraantennary oligosaccharides were found in trace amounts only. Immunohistochemical labeling of VE-cadherin and sialic acids displayed a codistribution along the intercellular junctions in endothelial cells of human umbilical arteries, veins, and cultured endothelial monolayers. Ca(2+)-depletion, performed on cultured endothelial cells, resulted in a reversible complete disappearance of VE-cadherin and of almost all sialic acid staining from the junctions. Sialidase treatment of whole cells caused a change of VE-cadherin immunofluorescence from a continuous and netlike superstructural organization to a scattered inconsistent one. Hence, cell surface sialic acids might play a role in VE-cadherin organization. PMID- 10460834 TI - Structure of the sulfated alpha-L-fucan from the egg jelly coat of the sea urchin Strongylocentrotus franciscanus: patterns of preferential 2-O- and 4-O-sulfation determine sperm cell recognition. AB - The egg jelly coats of sea urchins contains sulfated polysaccharides responsible for inducing the sperm acrosome reaction which is an obligatory event for sperm binding to, and fusion with, the egg. Here, we extend our study to the sea urchin Strongylocentrotus franciscanus. The egg jelly of this species contains a homofucan composed of 2- O -sulfated, 3-linked units which is the simplest structure ever reported for a sulfated fucan. This polysaccharide was compared with other sulfated alpha-L-fucans as inducers of acrosome reaction in conspecific and heterospecific sperm. Although all these fucans are linear polymers composed of 3-linked alpha-L-fucopyranosyl units, they differ in the proportions of 2-O- and 4-O-sulfation. The reactivity of the sperm of each species is more sensitive to the egg jelly sulfated fucan found in their own species. The reactivity of the sperm does not correlate with the charge density of the fucan, but with the proportion of 2-O- and 4-O-sulfation. The pattern of sulfation may be an important feature for recognition of fucans by the sperm receptor contributing to the species-specificity of fertilization. PMID- 10460835 TI - The formation of the oncofetal J28 glycotope involves core-2 beta6-N acetylglucosaminyltransferase and alpha3/4-fucosyltransferase activities. AB - The feto-acinar pancreatic protein or FAPP, the oncofetal glycoisoform of bile salt-dependent lipase (BSDL), is characterized by the presence of the J28 glycotope recognized by mAbJ28. This fucosylated epitope is carried out by the O linked glycans of the C-terminal mucin-like region of BSDL. This glycotope is expressed by human tumoral pancreatic tissues and by human pancreatic tumoral cell lines such as SOJ-6 and BxPC-3 cells. However, it is not expressed by the normal human pancreatic tissues and by MiaPaCa-2 and Panc-1 cells. Due to the presence of many putative sites for O-glycosylation on FAPP and BSDL, the structure of the J28 glycotope cannot be attained by classical physical methods. In the first part of the present study, we have determined which glycosyltransferases were differently expressed in pancreatic tumoral cell lines compared to normal tissues, focusing in part on fucosyltransferases (Fuc-T) and core-2 beta6-N-acetylglucosaminyltransferase (Core2GlcNAc-T). Our data suggested that alpha2-Fuc-T activity was decreased in the four cell lines tested (SOJ-6, BxPC-3, MiaPaCa-2, and Panc-1). The alpha(1-3) and alpha(1-4) fucosylations were decreased in tumor cells that do not express the J28 glycotope whereas alpha4-Fuc T and Core2GlcNAc-T activities were significantly increased in SOJ-6 cells which best expressed the J28 glycotope. Therefore, we wished to gain information about glycosyltransferases involved in the building of this structure by transfecting the cDNA encoding the mucin-like region of BSDL in CHO-K1 also expressing Core2GlcNAc-T and/or FUT3 and/or FUT7 activities. These CHO-K1 cells have been previously transfected with the cDNA encoding Core2GlcNAc-T and/or FUT3 and/or FUT7. Data indicated that the C-terminal peptide of BSDL (Cter) produced by those cells did not carry out the J28 glycotope unless Core2GlcNAc-T activity is present. Further transfection with FUT3 cDNA, increased the antibody recognition. Nevertheless, transfection with FUT3 or FUT7 alone did not generate the formation of the J28 glycotope on the C-terminal peptide. Furthermore, the Cter peptide produced by CHO-K1 cells expressing Core2GlcNAc-T was more reactive to the mAbJ28 after in vitro fucosylation with the recombinant soluble form of FUT3. These data suggested that the J28 glycotope encompasses structures initiated by Core2GlcNAc T and further fucosylated by alpha3/4-Fuc-T such as FUT3, likely on GlcNAc residues. PMID- 10460836 TI - Spatially and temporally regulated expression of N-acetylglucosamine-6-O sulfotransferase during mouse embryogenesis. AB - GlcNAc-6-O-sulfotransferase is involved in formation of 6-sulfo-N acetyllactosamine-containing structures such as 6-sulfo sialyl Lewis x. We investigated the mode of expression of GlcNAc-6-O-sulfotransferase during postimplantation embryogenesis in the mouse by in situ hybridization. Sulfotransferase mRNA was not detected on embryonic day (E) 6.5, while on E7.5 it was detected in the mesoderm, ectoderm, and ectoplacental cone. On E10.5, the sulfotransferase signals were mainly observed in the nervous tissue. On E12.5 and 13.5, various tissues in the process of differentiation expressed this mRNA. Several epithelial and mesenchymal tissues undergoing epithelial-mesenchymal interactions strongly expressed the mRNA. For example, in the developing tooth strong sulfotransferase mRNA expression was found only in the condensing mesenchyme on E13.5. On E13.5 and 15.5, the sites showing intense expression of the sulfotransferase again became restricted. In the brain, sulfotransferase mRNA was frequently found as discrete signals in narrow regions. These results suggest that 6-sulfo-N-acetyllactosamine structures have important roles in development. On E13.5 and 15.5, G152 (6-sulfo sialyl Lewis x antigen) was expressed in the neocortex, and AG223 (6-sulfo Lewis x antigen) in the thalamus and neocortex where the sulfotransferase signal was detected. However, in other organs, expression of these antigens did not correlate with the sulfotransferase mRNA, implicating complex nature of regulation of expression of the fucosyl 6-sulfo antigens. PMID- 10460837 TI - Requirement for a different hydrophobic moiety and reliable chromogenic substrate for endo-type glycosylceramidases. AB - A series of synthetic lactosides with aglycones that differed in length and structure were used to determine the substrate specificity of endo-type glycosylceramidases. Endoglycoceramidases (EGCase) from bacteria preferred lactosides with an acylamide structure over simple n-alkyl lactosides. While ceramide glycanase (CGase) from leech did not show preference. N -Acylaminoethyl beta-lactosides and n -alkyl lactosides were substrates for both EGCase and CGase, but N-acylaminobutyl beta-lactosides, whose acylamide residue differs from that in ceramide, were not hydrolyzed by EGCases. Thus, EGCases, but not CGase, appear to require an N-acyl group at the same position as that of intact glycosphingolipid for substrate recognition. A p-nitrophenyl lactoside derivative possessing an N-acyl chain was degraded by both EGCases and CGase and this chromogenic substrate may be an alternative substrate for endo-type glycosylceramidase activity. Km of the chromogenic lactoside for CGase and Rhodococcus EGCase were 28 microM and 2.9 mM, respectively. PMID- 10460852 TI - Herausgeber PMID- 10460853 TI - Krankenhaus agatharied: filmlos in die digitale zukunft PMID- 10460854 TI - BDR erreicht ruckwirkende anderungen zu Angio/Interventionen PMID- 10460855 TI - Niedersachsen paragraph signHVM-katastrophe in niedersachsen PMID- 10460856 TI - Leistungsverweigerung wegen kostenunterdeckung rechtswidrig. Aktuelles urteil des LSG nordrhein-westfalen PMID- 10460857 TI - Versichern sie sich uber den berufsverband !.Versicherungen zu spitzenkonditionen paragraph sign3. Berufshaftpflicht mit erweitertem straf-rechtschutz PMID- 10460858 TI - [Thorotrast-induced liver cancer: results of the German thorotrast study]. AB - AIMS: The X-ray contrast medium Thorotrast, used worldwide between 1930 to 1950 predominantly for arteriography, consisted of a colloidal solution of thorium dioxide. The radioactive thorium-232 (half-life 1.4x10(10) years) is stored lifelong in the organs of the reticulo-endothelial system after intravascular injection, causing chronic exposure to alpha radiation. The aim of the German Thorotrast study is the assessment of radiation late effects and the calculation of risk estimates. MATERIAL AND METHODS: The German Thorotrast study started in 1968 as a cohort study and comprises 2326 Thorotrast patients and 1890 patients of a matched control group. The Thorotrast patients who were still alive at the beginning of the study were examined by X-ray plain films of the upper abdomen and of the injection site of the contrast medium as well as by whole-body counter measurements. At the beginning we offered the patients ultrasound and later on CT and MRI at regular intervals for early detection of liver cancer. RESULTS: To date 454 primary liver cancers have been registered in the group of Thorotrast patients compared to 3 cases in the control group. With the help of modern imaging methods relatively small liver cancers were detected and could be surgically removed. DISCUSSION: There is a correlation between the mean accumulated dose to the liver and the incidence of liver cancer. The cumulative risk for liver malignancies is about 600 diseases per 10(4) persons whose liver was exposed to 1 Gy. Also the incidence of liver cirrhosis is correlated with the mean accumulated dose to the liver. PMID- 10460859 TI - [Computed tomography of the abdomen with multidetector-array CT]. AB - BACKGROUND: The advent of multidetector CT (MDCT) constitutes a quantum leap creating a wealth of new opportunities in medical imaging. However, while the basic principles of spiral CT still apply, we are now challenged to rethink our strategies in planning a CT study to take full advantage of the increased capabilities of MDCT. MATERIALS AND METHODS: We report here our preliminary experiences with MDCT for abdominal CT imaging within a 5-month period. During this time, suitable protocols for investigation of the abdomen using MDCT were developed. The capabilities of MDCT allow for tailoring dedicated CT protocols for routine applications as well as for biphasic liver studies and CT angiography of the aorta and abdominal vessels. RESULTS: The speed of MDCT can either be used to reduce the time needed to cover a given volume, or to use narrower beam collimations to increase the resolution of detail along the z-axis and reduce volume averaging. Higher scan speed allows reduction of the amount of contrast material in vascular applications and suppression of motion artifacts. Higher spatial resolution with thinner collimations reduces volume averaging and improves the detection of small hepatic and pancreatic lesions. Detailed analysis of vascular structures and high-quality three-dimensional reformations become feasible. New problems arise from the large amount of data generated by MDCT. CONCLUSIONS: MDCT offers a wealth of new opportunities that help us to come to a fast and accurate diagnosis in suspected abdominal disease. Traditional indications for performing CT are reemphasized and new clinical applications can be exploited. PMID- 10460860 TI - [New techniques and pulse sequences in MRI of the liver]. AB - PURPOSE: The MRI techniques which have contributed to increasing utilization of MRI for abdominal imaging are described and recent advances addressed. METHODS: For breath-hold examinations of the abdomen, two basic techniques are required: array coil technology and fast and ultrafast pulse sequences providing T1 and T2 contrast. RESULTS: Circular polarized array coils render high signal-to-noise ratios (SNR) within large imaging volumes. With fast gradient-echo sequences the liver can be scanned with or without fat saturation within one breath-hold. When adequate parameters are selected, T2-weighted fast (turbo) spin-echo sequences allow high contrast between normal liver tissue and focal liver lesions, even if breath-hold acquisition is applied. Moreover, good soft tissue contrast can also be achieved with ultrafast single-shot sequences. Based on this sequence type, MRCP with a 512 matrix could be performed. The "TRUE FISP" allows for high resolution visualisation of vessels without contrast media. Three-dimensional T1 weighted sequences allow for scanning the upper abdomen with a slice thickness of 3 mm within one breath-hold. Diffusion-weighted sequences contribute to the characterisation of focal liver lesions. DISCUSSION: Modern MRI technology including phase-array coils and high-performance gradient systems made it possible to perform all examinations in breath-hold techniques, reducing motion artifacts. PMID- 10460861 TI - [Multiphase, contrast-enhanced 3D-MR angiography for morphological and functional focal lesion detection. Initial results]. AB - PURPOSE: To investigate the value of multiphase breath-hold 3D gadolinium (Gd) enhanced MR angiography (MRA) for lesion detection and characterization of focal liver lesions. MATERIALS AND METHODS: Breath-hold 3D Gd-enhanced MRA was performed in 25 patients with benign and malignant hepatic lesions on a 1.5-T MR system using an ultrafast 3D spoiled gradient echo sequence (TR/TE =5/2 ms, FOV=300-450 mm, matrix=256x168, voxel volume=1.8x2.3 x2.5 mm, 64 partitions, central k-space reordering; acquisition time=27 s). Three measurements were done in the arterial, portal venous, and late venous phase. RESULTS: The analysis of the spatial and temporal evolution of contrast enhancement of the 3D-MRA improved significantly (P<0.01) lesion detection and characterization if compared with T1 precontrast, T2-weighted, and T1 postcontrast images. CONCLUSION: Multiphase breath-hold 3D Gd-enhanced MRA imaging is a robust new technique to significantly improve morphological detection of benign and malignant lesions during the early arterial phase and further improves functional characterization of liver lesions by a combination of an arterial, a portal venous, and a late venous phase. Schlussselworter Multiphasisch. MR-Angiographie. Leberlasionen PMID- 10460862 TI - [Comparison of dual-phase helical CT with native and ferum oxide-enhanced magnetic resonance imaging in detection and characterization of focal liver lesions]. AB - PURPOSE: Evaluation of the diagnostic efficacy of contrast-enhanced, dual-phase spiral CT and MRI before and after administration of SPIO particles in focal hepatic disease with previously uncertain diagnosis. MATERIAL AND METHODS: In 46 patients in whom primary or secondary hepatic malignancy was suspected, dual phase spiral CECT and breath-held T1-weighted gradient-echo and T2-weighted fast spin-echo MRI (1.5 T, body-phased-array coil) before and after SPIO administration were compared. The indications for the subsequent MRI studies were based on ambiguous findings on CECT. The number of hepatic lesions, the overall lesion detection and characterization was evaluated by consensus and compared to the gold standard (histological proof in 30/46 of the cases, long-term follow-up in 16/46 of the cases). RESULTS: In 34 of 46 cases the correct diagnoses were established by CECT (sensitivity 96%, specificity 48%) revealing significantly less lesions than MRI. Unenhanced MRI had sensitivity of 93% and specificity of 71%, whereas SPIO-enhanced MRI had sensitivity of 97% and specificity of 88%. The differences between the modalities were even more pronounced in the detection of lesions smaller than 10 mm with SPIO-MRI as the most sensitive method. CONCLUSIONS: In this problem-oriented scenario, SPIO-enhanced MRI was superior to spiral CT and unenhanced MRI regarding the diagnostic efficacy in the pre operative work-up of focal liver lesions. SPIO-enhanced MRI can be recommended as a problem-solving tool for the clinical routine. PMID- 10460863 TI - [Magnetic resonance imaging of benign and premalignant tumors in childhood]. AB - PURPOSE: To evaluate the potential of MRI in determining benign and premalignant abdominal tumors in childhood. METHODS: MR images of 93 children with 69 malignant and 24 benign abdominal tumors were analyzed retrospectively without and with knowledge of clinical findings. Based on the final diagnosis, MR findings were surveyed with regard to the correct differential diagnosis and to the differentiation between benign and malignant masses. RESULTS: Analysis of MRI alone revealed relatively low sensitivity of 67% for diagnosis of a benign tumor. The main reason was unspecific morphologic criteria leading to the false-negative diagnosis of a malignant tumor. Together with clinical and laboratory findings, sensitivity could be increased to 92%. The main criterion for differentiation of malignant tumors turned to be out the origin of tumor. Benign tumors could be best differentiated by their internal structure. CONCLUSION: When considering clinical findings in pediatric abdominal tumors, MRI represents a valuable modality for differential diagnosis. PMID- 10460864 TI - [Echo-planar diffusion-weighted MRI in the diagnosis of acute ischemic stroke: characterisation of tissue abnormalities and limitations in the interpretation of imaging findings]. AB - INTRODUCTION: An accurate diagnosis is frequently difficult in early stroke. Diffusion-weighted (DW) magnetic resonance imaging (MRI) allows visualization of ischemic parenchyma and quantitative assessment of tissue changes before unequivocal abnormalities appear on T(2)-weighted MRI. METHODS: We analyzed 105 MRI examinations of patients with acute stroke (<24 h) with regard to patterns of abnormalities in T(2)-weighted and DW MRI. Furthermore we assessed the influence of artifacts related to DW echo-planar single-shot MRI on image interpretation. RESULTS: Depending on the time of patient assessment there were three partly overlapping T(2)/DW patterns: (1) in the very early phase (/=3 h). Typical artifacts (susceptibility distortions, N/2 artifact, chemical shift artifact and eddy currents artifact) had to be considered when interpreting images. CONCLUSIONS: Provided typical artifacts are taken into consideration, echo-planar DW MRI allows a more precise diagnostic assessment in acute stroke. PMID- 10460865 TI - [A case of unusual space-occupying lesion in the knee joint]. PMID- 10460866 TI - [Digital projection radiography. Technical bases. Imaging quality and usefulness]. PMID- 10460867 TI - Hyaluronidase ameliorates rejection-induced edema. AB - Hyaluronan, a glucosaminoglycan with unique water-binding capacity, is accumulated in the interstitial edematous tissue in rejecting organs. We here investigated whether the increased tissue content of water and hyaluronan seen during allograft rejection can be prevented by treatment with the hyaluronan degrading enzyme hyaluronidase. Heterotopic heart transplantations between PVG and Wistar/Kyoto rats were performed. Recipient rats were treated with hyaluronidase prophylactically or therapeutically, either alone or in combination with cyclosporine. Daily intravenous injections of hyaluronidase induced a significant reduction of the cardiac content of both hyaluronan and water, as evaluated on day six after transplantation. Morphological examination revealed grafts with better preserved morphology and fewer infiltrating mononuclear cells, compared to untreated controls. Hyaluronidase therapy, alone or combined with cyclosporine, resulted in prolonged graft survival times. Hyaluronidase infusion for two hours also reduced already established edema five days after transplantation. This study confirms the hypothesis that hyaluronan accumulation plays a critical role in edema formation, and that hyaluronidase therapy can be used to reduce edema after organ transplantation. PMID- 10460868 TI - Permanently reduced plasma ionized magnesium among renal transplant recipients on cyclosporine. AB - Hypomagnesemia is common after kidney transplantation. Until recently, only the determination of total plasma magnesium was possible, whereas the assessment of ionized magnesium has since become practicable. One hundred and nine renal transplant patients on cyclosporine with allografts functioning stably for more than 6 months and plasma creatinine levels of less than 200 micromol/l entered the study. Total and ionized circulating magnesium were assessed among these 109 patients, as well as among 15 renal transplant patients not on cyclosporine and 21 healthy volunteers. Cyclosporine patients showed significantly lower total and ionized circulating magnesium values than the two control groups. Plasma total and ionized magnesium levels were also significantly lower among cyclosporine patients treated concurrently with insulin or oral hypoglycemic agents than among those who were not. No correlation was noted between time after transplantation and plasma magnesium with respect to patients on cyclosporine. In conclusion, the study demonstrates that a large subset of renal transplant patients treated with cyclosporine have permanent deficiencies of ionized and total magnesium. The tendency towards hypomagnesemia is also more pronounced among patients with diabetes mellitus. PMID- 10460869 TI - T-cell immune defect and B-cell activation in renal transplant recipients with monoclonal gammopathies. AB - Monoclonal immunoglobulins (molg) have repeatedly been described in organ and bone marrow transplantation. Although their exact significance is not known, their occurrence is often associated with intensive immunosuppression. We investigated whether molg reflect T-cell immune defect and B-cell activation in renal transplant recipients. Immunofixations and lymphocyte subset analysis (CD4, CD8, CD19) were performed in 182 renal transplant recipients. Soluble CD23 concentrations were measured in patients with molg and in control transplant patients without molg. Monoclonal immunoglobulins were identified in 54 patients (29.6 %). Transplant endurance was shorter (62 +/- 53 months vs 81 +/- 47 months; P < 0.02) and age was older (53 +/- 13 years vs 46 +/- 13 years; P < 0.005) in patients with molg. Maintenance immunosuppression did not differ between patients with and without molg. Mean CD4-cell count was significantly lower in patients with molg (387 +/- 286/mm(3) vs 538 +/- 341/mm(3); P < 0.005). Both CD8- and CD19 cell counts were similar for the 2 groups. Soluble CD23 concentrations were higher in patients with abnormal immunoglobulin values than in patients with normal immunofixation (12.8 +/- 8 vs 1.9 +/- 1.8 microg/l; P < 0. 005). Our study provides new evidence that molg reflect T-cell immune defect in renal transplant recipients. Further studies are required to determine whether CD4-cell count and sCD23 may help to predict the risk of lymphoma in transplant patients with molg. PMID- 10460870 TI - Intravenous ganciclovir prophylaxis for cytomegalovirus in heart, heart-lung, and lung transplant recipients. AB - Cytomegalovirus (CMV) disease has had a significant clinical impact on the heart, heart-lung and lung transplant recipients in our centre. CMV disease has been so severe with CMV antibody-negative heart-lung transplant patients receiving organs from CMV antibody-positive donors (CMV-mismatched patients) that in 1986 we adopted the policy of not transplanting CMV-positive organs into CMV-negative heart-lung or lung recipients. In December 1992, we instituted a policy of providing intravenous ganciclovir (5 mg/kg twice a day for 28 days) during the immediate postoperative period for CMV-mismatched heart recipients and CMV antibody-positive heart-lung and lung patients, who have been the patients at greatest risk of severe CMV disease in our centre. A placebo group was not employed because of ethical considerations, ganciclovir having been shown to be effective for the treatment of CMV infections among transplant patients. Compared with a historical control group of patients receiving no prophylaxis, prophylactic ganciclovir reduced the incidence of CMV infection (39 % vs 91 %, P = 0.0006) and CMV disease (17 % vs 74 %, P = 0.0004) among CMV antibody-positive heart-lung recipients. Prophylactic ganciclovir did not significantly reduce the incidence of CMV infection or disease among heart or isolated lung recipients. Ganciclovir was well tolerated, with few adverse reactions. In the case of heart lung transplant patients, one month of intravenous prophylactic ganciclovir significantly reduced the incidence of both CMV infection and disease when compared with patients who received no prophylaxis. With the lung transplant and heart transplant patients, there were no significant differences between the prophylaxis and nonprophylaxis groups, although there was a consistent trend towards less infection and disease in the prophylaxis groups. PMID- 10460871 TI - Local liberation of cytokines during liver preservation. AB - In order to investigate locally produced mediators during the process of organ storage in liver transplantation, we collected the liver preservation solution effluent of 15 transplanted livers and compared it with serum samples taken preoperatively from donor and recipient, as well as 60 min after reperfusion. The mean ischemia time +/- SEM was 10 h 10 min +/- 53 min. Mean concentrations in University of Wisconsin preservation solution effluent were: interleukin-(IL )1beta 154 +/- 77 pg/ml; IL-1 receptor antagonist (IL-1 ra) 1281 +/- 309 pg/ml; IL-6 412 +/- 90 pg/ml; and for tumor necrosis factor-(TNF-)alpha 74 +/- 21 pg/ml. Cytokine levels in the donors were lower than those detected in the effluent. All measured cytokines showed higher concentrations in the effluent compared to those of the recipient prior to the operation. With respect to a comparison of donor and recipient values, no correlation is evident. Likewise, the ischemic time does not correlate with effluent values. Further development of liver preservation concepts requires information about the state of the graft before reperfusion. Data on cytokine liberation may serve as a helpful tool for the further development of preservation concepts because they enable an estimation of cell activation during preservation. PMID- 10460872 TI - Eosinophilic alveolitis in BAL after lung transplantation. AB - Lung transplantation has become a therapeutic option for patients with end stage lung disease. However, outcome after transplantation is complicated by episodes of rejection and infections. Bronchoalveolar lavage is a valuable tool in monitoring patients after transplantation, since it allows the detection of pathogens. A marker specifically indicating rejection from changes in BAL fluid has not been found yet. Especially changes in differential cell count, like lymphocytosis or an increase in polymorphnuclear granulocytes, are unspecific. The role of high eosinophil levels in BAL has not been elucidated yet. We analyzed 25 BAL samples and clinical data of 4 patients who underwent lung transplantation and presented with recurrent episodes of eosinophilic alveolitis in BAL. All patients demonstrated a deterioration of clinical condition, lung function, and blood gas analysis during times of eosinophilia in BAL, compared to previous examinations. In all cases, eosinophilia in BAL was accompanied by rejection. All patients were finally treated with high doses of steroids, resulting in improvement of all parameters. Eosinophilia was not associated with significant changes in the IL-5 concentration in BAL or the pattern of IL-5 expression in BAL cells. In conclusion, eosinophilic alveolitis may indicate acute rejection in patients after lung transplantation, if other causes of eosinophilia are excluded. PMID- 10460873 TI - Successful recanalization of late portal vein thrombosis after liver transplantation using systemic low-dose recombinant tissue plasminogen activator. AB - Portal vein thrombosis (PVT) is an infrequent complication following hepatic transplantation. However, deterioration of liver function and accompanying complications may be life threatening. Several attempts of surgical or percutaneous transhepatic procedures have been described. In some cases high dose fibrinolytic regimens have been successful. We describe the case of a male liver recipient with recurrent liver fibrosis due to hepatitis B reinfection and late portal vein thrombosis 45 months after transplantation. Complete recanalization was achieved using systemic low dose recombinant tissue plasminogen activator (rt PA). PMID- 10460874 TI - Transmission of factor VII deficiency through liver transplantation. AB - The liver is the primary site of synthesis for the majority of coagulation factors. There are published accounts of liver donor-to-recipient transmission of protein C deficiency with dysfibrinogenemia and factor XI deficiency. In this article, we report what we believe to be the first observation, of transmission of factor VII deficiency, a rare, autosomal recessive coagulation disorder, from an affected liver donor to a naive liver recipient. At 300 days after transplantation, the recipient remains with an isolated prolongation of the prothrombin time and a below-normal level of factor VII, and has had no bleeding complications. PMID- 10460875 TI - Successful transplantation of pediatric en bloc kidneys with bilateral double ureters. AB - We report a case in which en bloc kidneys with bilateral double ureters from a 5 month-old donor were successfully transplanted into a 25-year-old recipient. No stents were used. There were no complications after the transplant. The patient remains well at more than 1.5 years post-transplantation with serum creatinine 1.2 mg/dl. PMID- 10460876 TI - Disappearance of hepatitis B virus core deletion mutants and successful combined kidney/liver transplantation in a patient treated with lamivudine. AB - Hepatitis B virus (HBV) core deletion variants with enhanced viral replication are associated with rapid deterioration of liver function in renal allograft recipients. Antiviral agents such as famciclovir and lamivudine offer new treatment strategies for these patients. Appearance, accumulation and persistence of HBV core deletion mutants were closely monitored in a kidney transplant recipient with liver cirrhosis before and after initiation of antiviral treatment. Under treatment with famciclovir HBV DNA concentration decreased by 50 %, HBV mutants persisted. After replacement of famciclovir by lamivudine HBV replication was reduced below the detection limit. Lamivudine was well tolerated and liver function improved. After successful combined kidney/liver transplantation the patient became HBsAg and HBV DNA (detected by PCR) negative under continuous hyperimmune globulin and lamivudine treatment. Antiviral therapy with lamivudine may be useful in treatment of progressive liver disease associated with HBV core deletion mutants in renal allograft recipients and may enable successful liver transplantation. PMID- 10460877 TI - Conversion from cyclosporin (Neoral) to tacrolimus (Prograf) in renal allograft recipients with chronic graft nephropathy: results of an observational study. AB - To evaluate the role of tacrolimus in the treatment of Chronic Graft Nephropathy (CGN), a pilot cross-sectional study was performed on 14 patients with deteriorating renal function and biopsy-proven CGN. Maintenance therapy was switched from cyclosporin to tacrolimus, and results of conversion on allograft function were assessed by estimated glomerular filtration rate (GFR) and clinical outcome. Minimum follow-up was 15 months. Two distinctive response patterns emerged: (i) continuing deterioration of renal function with no apparent benefit over the projected trend of GFR (nine patients), and (ii) unequivocal change in the GFR trend line equation with reduced rate of deterioration in one patient and sustained improvement of GFR in four patients (reversal of downward trend). Five out of 14 patients (36 %) benefited from replacing Neoral with Prograf. All five patients exceeded their estimated time of return to dialysis by a median of 41 weeks (range: 29-52) and their grafts continue to function. PMID- 10460878 TI - Malakoplakia of the caecum in a kidney-transplant recipient: presentation as acute tumoral perforation and fatal outcome. AB - Malakoplakia is a rare pseudotumoral inflammatory disease known to affect immunocompromised subjects, mainly with a history of recurrent Escherichia coli infection. The urinary tract is the most frequent site of the disease, although all organs can be involved. In the present article, we report a case of malakoplakia of the caecum, that developed in a 52-year-old man, who had received a kidney transplant 9 years before and had a history of recurrent E. coli urinary tract infections. Malakoplakia presented as acute intestinal perforation, and, despite aggressive surgical and medical management, disease progressed toward a fatal outcome due to sepsis and multiple organ failure 9 months later. A defect in the macrophagic activity was demonstrated. PMID- 10460879 TI - Gastrointestinal mucormycosis and liver transplantation; a case report and review of the literature. PMID- 10460880 TI - The new Eurotransplant kidney allocation system: a justified balance between equity and utility? PMID- 10460881 TI - Upcoming meetings and workshops PMID- 10460882 TI - Announcements PMID- 10460883 TI - Recent advances in understanding resin acid biodegradation: microbial diversity and metabolism. AB - Resin acids are tricyclic diterpenoids that are found in the oleoresin of coniferous trees. Resin-acid-degrading microorganisms are ubiquitous in the environment. The bacterial isolates that grow on resin acids as sole organic substrates are physiologically and phylogenetically diverse, and include psychrotolerant, mesophilic, and thermophilic bacteria. Recent studies of the biodegradation of resin acids by these organisms have demonstrated that in gram negative bacteria, distinct biochemical pathways exist for the degradation of abietane- and pimerane-type resin acids. One of these organisms, Pseudomonas abietaniphila BKME-9, harbors a convergent pathway that channels the nonaromatic abietanes and dehydroabietic acid into 7-oxodehydroabietic acid. This dioxygenolytic pathway is encoded by the recently cloned and sequenced dit gene cluster. The dit cluster encodes the ferredoxin and the alpha- and beta-subunits of a new class of ring-hydroxylating dioxygenases as well as an extradiol ring cleavage dioxygenase. Although it was previously thought that resin acids are very recalcitrant under anoxic conditions, recent investigations have demonstrated that they are partially metabolized under anoxic conditions by undefined microorganisms. The anaerobic degradation of resin acids principally generates aromatized and decarboxylated products (such as retene) that are thought to persist in the environment. PMID- 10460884 TI - Isolation and characterization of a Clostridium sp. with cinnamoyl esterase activity and unusual cell envelope ultrastructure. AB - Microorganisms that hydrolyse the ester linkages between phenolic acids and polysaccharides in plant cell walls are potential sources of enzymes for the degradation of lignocellulosic waste. An anaerobic, mesophilic, spore-forming, xylanolytic bacterium with high hydroxy cinnamic acid esterase activity was isolated from the gut of the grass-eating termite Tumilitermes pastinator. The bacterium was motile and rod-shaped, stained gram-positive, had an eight-layered cell envelope, and formed endospores. Phylogenetic analysis based on 16S rRNA indicated that the bacterium is closely related to Clostridium xylanolyticum and is grouped with polysaccharolytic strains of clostridia. A wide range of carbohydrates were fermented, and growth was stimulated by either xylan or cellobiose as substrates. The bacterium hydrolysed and then hydrogenated the hydroxy cinnamic acids (ferulic and p-coumaric acids), which are esterified to arabinoxylan in plant cell walls. Three cytoplasmic enzymes with hydroxy cinnamic acid esterase activity were identified using non-denaturing gel electrophoresis. This bacterium possesses an unusual multilayered cell envelope in which both leaflets of the cytoplasmic membrane, the peptidoglycan layer and the S layer are clearly discernible. The fate of all these components was easily followed throughout the endospore formation process. The peptidoglycan component persisted during the entire morphogenesis. It was seen to enter the septum and to pass with the engulfing membranes to surround the prespore. It eventually expanded to form the cortex, verification for the peptidoglycan origin of the cortex. Sporogenic vesicles, which are derived from the cell wall peptidoglycan, were associated with the engulfment process. Spore coat fragments appeared early, in stage II, though spore coat formation was not complete until after cortex formation. PMID- 10460885 TI - Aerobic turnover of dimethyl sulfide by the anoxygenic phototrophic bacterium thiocapsa roseopersicina AB - This is the first report describing the complete oxidation of dimethyl sulfide (DMS) to sulfate by an anoxygenic, phototrophic purple sulfur bacterium. Complete DMS oxidation was observed in cultures of Thiocapsa roseopersicina M11 incubated under oxic/light conditions, resulting in a yield of 30.1 mg protein mmol(-1). No oxidation of DMS occurred under anoxic/light conditions. Chloroform, methyl butyl ether, and 3-amino-1,2,4-triazole, which are specific inhibitors of aerobic DMS oxidation in thiobacilli and hyphomicrobia, did not affect DMS oxidation in strain M11. This could be due to limited transport of the inhibitors through the cell membrane. The growth yield on sulfide as sole electron donor was 22.2 mg protein mmol(-1) under anoxic/light conditions. Since aerobic respiration of sulfide would have resulted in yields lower than 22 mg protein mmol(-1), the higher yield on DMS under oxic/light conditions suggests that the methyl groups of DMS have served as an additional carbon source or as an electron donor in addition to the sulfide moiety. The kinetic parameters V(max) and K(m) for DMS oxidation under oxic/light conditions were 12.4 +/- 1.3 nmol (mg protein)(-1) min(-1) and 2 &mgr;M, respectively. T. roseopersicina M11 also produced DMS by cleavage of dimethylsulfoniopropionate (DMSP). Specific DMSP cleavage rates increased with increasing initial substrate concentrations, suggesting that DMSP lyase was only partly induced at lower initial DMSP concentrations. A comparison of T. roseopersicina strains revealed that only strain M11 was able to oxidize DMS and cleave DMSP. Both strain M11 and strain 5811 accumulated DMSP intracellularly during growth, while strain 1711 showed neither of these characteristics. Phylogenetic comparison based on 16S rRNA gene sequence revealed a similarity of 99.0% between strain M11 and strain 5811, and 97.6% between strain M11 and strain 1711. DMS and DMSP utilization thus appear to be strain specific. PMID- 10460886 TI - Sexual development of Aspergillus nidulans in tryptophan auxotrophic strains. AB - The interplay between sexual development and amino acid biosynthesis in the ascomycete Aspergillus nidulans was studied. The growth and differentiation of four tryptophan auxotrophic strains that are unable to regulate their tryptophan biosynthesis, were examined. These strains are sterile on medium containing low tryptophan concentrations. Fruitbody formation was restored by supplementation with high concentrations of tryptophan and was promoted by supplementation with indole or auxin. Tryptophan supplementation resulted in auxin production of A. nidulans. Fertility of ascospores of the tryptophan auxotrophic strains could only be partially re-established. trpC transcript levels and enzyme activities remained stable in cleistothecia and conidiophore extracts as compared to those of mycelium, while levels of gene transcripts involved in glycolysis were lower in fruitbodies and conidiospores. Auxotrophic strains unable to form fruitbodies at intermediate amino acid supplementation levels turned on the cross-pathway regulatory system that is induced by amino acid starvation. We conclude that there is a connection between the genetic network of cross-pathway control and sexual development in A. nidulans. PMID- 10460887 TI - Heliorestis daurensis, gen. nov. Sp. Nov., An alkaliphilic rod-to-coiled-shaped phototrophic heliobacterium from a siberian soda lake AB - A novel alkaliphilic heliobacterium was isolated from microbial mats of a low salt alkaline Siberian soda lake. Cells of the new organism were tightly coiled when grown in coculture with a rod-shaped bacterium, but grew as short filaments when finally obtained in pure culture. The new phototroph, designated strain BT H1, produced bacteriochlorophyll g and a neurosporene-like pigment, and lacked internal photosynthetic membranes. Similar to other heliobacteria, strain BT-H1 grew photoheterotrophically on a limited range of organic compounds including acetate and pyruvate. Sulfide was oxidized to elemental sulfur and polysulfides under photoheterotrophic conditions; however, photoautotrophic growth was not observed. Cultures of strain BT-H1 were alkaliphilic, growing optimally at pH 9, and unlike other heliobacteria, they grew optimally at a temperature of 25 degrees C rather than at 40 degrees C or above. Analysis of the 16S rRNA gene sequence of the new organism showed that it groups within the heliobacterial clade. However, its branching order was phylogenetically basal to all previously investigated species of heliobacteria. The G+C content of the DNA of strain BT-H1 (44.9 mol%) was also quite distinct from that of other heliobacteria. This unique assemblage of properties implicates strain BT-H1 as a new genus and species of the heliobacteria, Heliorestis daurensis, named for its unusual morphology ("restis" is Latin for "rope") and for the Daur Steppe in Russia in which these soda lakes are located. PMID- 10460888 TI - Purification of cold-shock-like proteins from Stigmatella aurantiaca - molecular cloning and characterization of the cspA gene. AB - Prominent low-molecular-weight proteins were isolated from vegetative cells of the myxobacterium Stigmatella aurantiaca and were found to be members of the cold shock protein family. A first gene of this family (cspA) was cloned and sequenced. It encodes a protein of 68 amino acid residues that displays up to 71% sequence identity with other bacterial cold-shock(-like) proteins. A cysteine residue within the RNP-2 motif is a peculiarity of Stigmatella CspA. A cspA::(Deltatrp-lacZ) fusion gene construct was introduced into Stigmatella by electroporation, a method that has not been used previously for this strain. Analysis of the resultant transformants revealed that cspA transcription occurs at high levels during vegetative growth at 20 and 32 degrees C, and during fruiting body formation. PMID- 10460889 TI - Purification and characterization of 5-oxo-L-prolinase from Paecilomyces varioti F-1, an ATP-dependent hydrolase active with L-2-oxothiazolidine-4-carboxylic acid. AB - An enzyme cleaving L-2-oxothiazolidine-4-carboxylic acid to L-cysteine was purified 75-fold with 8% recovery to near homogeneity from crude extracts of Paecilomyces varioti F-1, which had been isolated as a fungus able to assimilate L-2-oxothiazolidine-4-carboxylic acid. The molecular mass was estimated to be 260 kDa by gel filtration. The purified preparation migrated as a single band of molecular mass 140 kDa upon SDS-PAGE. The maximum activity was observed at a range of pH 7.0-8.0 and at 50 degrees C. The enzyme activity was completely inhibited by SH-blocking reagents such as AgNO(3), p-chloromercuribenzoic acid, N ethylmaleimide, and N-bromosuccinimide. The enzyme required ATP, Mg(2+), and KCl for the cleavage of L-2-oxothiazolidine-4-carboxylic acid. The enzyme also cleaved 5-oxo-L-proline to L-glutamic acid and is considered to be 5-oxo-L prolinase. PMID- 10460890 TI - Quantification of osteopontin in the urine of healthy and stone-forming men. AB - Osteopontin (OPN) is one of the most important components in calcium stone matrix, but its role in stone formation is not clear. Since quantitative data regarding the excretion of OPN are necessary to assess its role, we have developed a quantitative enzyme-linked immunosorbent assay (ELISA) for OPN, and measured the urinary OPN concentrations in urolithiasis patients. Forty-seven men with urinary stones composed chiefly of calcium oxalate participated in the study. The controls were 13 normal healthy male volunteers. Urine samples were collected early in the morning and analyzed by a quantitative ELISA employing purified polyclonal antibodies to synthesized OPN aminopolypeptides. The urinary ratio of the concentrations of OPN and creatinine (OPN/Cre) in the urolithiasis patients (0.039 +/- 0.029) was significantly lower than that in the control subjects (0.062 +/- 0.030) (P<0.05). Single stone formers (n = 26; 0.050 +/- 0.020) had significantly higher OPN/Cre ratios compared with recurrent stone formers (n = 21; 0.031 +/- 0.021) (P<0. 05). The results show that OPN excretion in urolithiasis patients was lowered, presumably because of the incorporation of OPN by kidney stones. PMID- 10460891 TI - Calcium oxalate crystallization kinetics at different concentrations of human and artificial urine, with a constant calcium to oxalate ratio. AB - The effect of in vitro dilution of artificial urine or human urine on the crystallization of calcium oxalate was examined in a mixed suspension, mixed product removal crystallization system. Direct growth inhibition by components of artificial urine was not significant and supersaturation was the dominant factor in determining crystal nucleation and growth rates. Dilution of human urine caused a decrease in crystal growth rate that was independent of the input calcium and oxalate concentrations, suggesting that dilution of growth inhibitors could be physiologically more important than any reduction in supersaturation. This loss of growth inhibition was counteracted by a reduction in nucleation promotion, with the net effect that the mass of crystals declined. Correlation of crystallization measurements with urinary concentration (osmotic pressure) confirmed these observations, with a negative relationship for growth rate and a positive relationship for nucleation rate and suspension density. Increasing the concentration of urine shifts the crystallization balance from low nucleation/high growth to high nucleation/low growth. Calcium oxalate crystalluria in healthy urine is therefore less likely at early stages of urine development in the nephron and the likelihood can be further reduced by increased fluid output. Our results suggest that lowering the heterogeneous nucleation activity by dilution is more than sufficient to override the loss of growth inhibition. PMID- 10460892 TI - Distribution of osteopontin and calprotectin as matrix protein in calcium containing stone. AB - We recently reported that osteopontin (OPN) and calprotectin (CPT) are present in the matrix of urinary calcium stones, and that OPN mRNA is expressed in the renal distal tubular cells. In the present study, we examined the immunohistochemical distributions of OPN and CPT in urinary stones. The stones used in this study were passed spontaneously from the upper urinary tract. One half of each of the stones was analyzed with an infrared spectrophotometer, and were shown to be comprised of calcium oxalate, calcium phosphate, uric acid and cystine. The other half of each stone was immersed in tetrasodium ethylenediamine-tetraacetate (EDTA) solution. The half-stones were embedded in paraffin and cut into 5-microm sections. The avidin-biotin-peroxidase complex technique was employed. A monoclonal antibody to human milk-derived OPN and a monoclonal antibody to human granulocyte-derived CPT were used as primary antibodies. The immunochemical study using the OPN and CPT antibodies showed positive staining of the matrix of the urinary calcium stones. The stones showed staining in two distinct zones: a core area was stained with randomly aggregated OPN and CPT, and peripheral layers were stained in concentric circles. On the basis of our observations, it is reasonable to presume that OPN and CPT play roles as the matrix in the structure of urinary calcium stones. PMID- 10460893 TI - Alpha-1-microglobulin: inhibitory effect on calcium oxalate crystallization in vitro and decreased urinary concentration in calcium oxalate stone formers. AB - In the past few years, alpha-1-microglobulin (alpha1m) has been copurified from human urine with bikunin, a potent inhibitor of calcium oxalate (CaOx) crystallization in vitro. In this study, we have purified alpha1m without bikunin contamination and investigated its possible role in CaOx crystallization by in vitro and in vivo studies. Alpha-1m was purified with an anti-alpha1m antibodies CNBr-activated sepharose column. Two molecular species of alpha1m of respectively 30 and 60 kDa were purified. For each protein, two blots of 30 and 60 kDa cross reacted with anti-alpha1m antibodies, suggesting that these two forms were derived one from the other. Both protein species inhibited CaOx crystallization in a dose-dependent manner in two in vitro tests. In the first test, the presence of alpha1m of 30 kDa (8 microg/ml) in a medium containing 0. 76 mM CaCl(2) (with (45)Ca) and 0.76 mM Ox(NH(4))(2) inhibited CaOx crystallization by 38% as estimated by supernatant radioactivity after 1 h of agitation. In the second test, CaOx kinetics were examined for 3 to 10 min in a turbidimetric model at 620 nm. The presence of alpha1m of 30 kDa in a medium containing 4 mM CaCl(2) and 0.5 mM Na(2)Ox inhibited CaOx crystallization by 41.5%, as estimated by the slope modification of turbidimetric curve. Alpha-1m can be considered as another inhibitor of urinary CaOx crystal formation, as shown by the present in vitro studies. Using an ELISA assay, we found that urinary alpha1m concentration was significantly lower in 31 CaOx stone formers than in 18 healthy subjects (2.95 +/ 0.29 vs 5.34 +/- 1.08 mg/l respectively, P = 0.01). The decreased concentration of alpha1m in CaOx stone formers could be responsible in these patients, at least in part, for an increased risk of CaOx crystalluria. PMID- 10460894 TI - Isoelectric focusing of native urinary uromodulin (Tamm-Horsfall protein) shows no physicochemical differences between stone formers and non-stone formers. AB - Recent studies have suggested the occurrence of an abnormal form of uromodulin in stone formers which may be related to a reduced sialic acid content of the protein in these patients. Previous attempts to demonstrate these differences have required extensive sample processing prior to analysis, which may in itself alter the nature and behaviour of uromodulin. By employing a sensitive detection system of enhanced chemiluminescence on Western blots following isoelectric focusing we have been able to study the physicochemical properties of uromodulin in whole unprocessed urine from 50 idiopathic calcium stone formers and 15 non stone formers. Uromodulin, desialated in vitro with either acid or neuraminidase was also analysed using the same system. All urine samples analysed from stone formers and non-stone formers showed a single band of pI 3.5 after isoelectric focusing. Desialated uromodulin showed a series of bands ranging from pI 4.0 to 5.1 reflecting different amounts of sialic acid removed. We conclude there are no charge-related differences in native uromodulin between stone formers and non stone formers, in particular none relating to the sialic acid content of the protein. PMID- 10460896 TI - The impact of the geometry of the lithotriptor aperture on fragmentation effect at extracorporeal shock wave lithotripsy treatment. AB - The aim of this study was to determine the impact of the aperture size of an electro-hydraulic lithotriptor on the fragmentation effect. We also wanted to investigate whether a potential change in the capacitance of the pulse forming network (PFN), at a certain energy level, might have an impact on fragmentation rate. Two different apertures with a diameter of 23 and 17 cm respectively were compared using two different values of total PFN capacitance: 50 nF and 80 nF. Model stones of similar size and weight were fragmented. The number of shots for complete fragmentation or the grade of fragmentation after a certain number of shots was measured. This study shows that for the shock wave system used, the 23 cm aperture seems to provide more effective fragmentation as function of the number of shots compared with the 17-cm aperture at the same energy level. Furthermore, a minor change in the PFN capacitance between reasonable limits does not affect the fragmentation efficiency. This article also highlights the fact that it is not relevant simply to compare the voltage level given in the shots in extracorporeal shock wave lithotripsy treatment between different lithotriptors. PMID- 10460895 TI - Expression of bone matrix proteins in urolithiasis model rats. AB - Urinary calcium stones are a pathological substance, and they show similarities to physiological mineralization and other pathological mineralizations. The expression of messenger (m) RNAs of osteopontin (OPN), matrix Gla protein (MGP), osteonectin (ON) and osteocalcin (OC) in bones and teeth has been described. We previously identified OPN as an important stone matrix protein. In addition, the spontaneous calcification of arteries and cartilage in mice lacking MGP was recently reported, a finding which indicates that MGP has a function as an inhibitor of mineralization. Here, we examined the mRNA expressions of OPN, MGP, ON, and OC in the kidneys of stone-forming model rats administered an oxalate precursor, ethylene glycol (EG) for up to 28 days. The Northern blotting showed that the mRNA expressions of OPN and MGP were markedly increased with the administration of EG, but their expression patterns differed. The OPN mRNA expression reached the maximal level at day 7 after the initiation of the EG treatment and showed no significant difference after 14 and 28 days, whereas the MGP mRNA expression rose gradually to day 28. The in situ hybridization demonstrated that the cell type expressing OPN mRNA was different from that expressing MGP. We suggest that OPN acts on calcification and MGP acts on suppression. PMID- 10460897 TI - How to improve lithotripsy and chemolitholysis of brushite-stones: an in vitro study. AB - Because of their resistance to fragmentation, treatment of brushite stones is a big problem. This study was performed to look for an improvement in therapeutic strategies by using artificial stones made of brushite (Bon(n)-stones), which are comparable to their natural counterparts. Using an ultrasound transmission technique, longitudinal wave propagation speeds were measured at different time intervals during treatment with hemiacidrin. From these and density measurements, transverse wave speed, wave impedance and dynamic mechanical properties of the artificial stones were calculated. Moreover, the microhardness of artificial stones was measured and investigations on shock wave lithotripsy (SWL) combined with initial chemolytic treatment of the stones were performed. The suggestion was confirmed that stone fragility and thus SWL can be improved by varying the physical properties of brushite stones through treatment with hemiacidrin. Additionally, we demonstrated the efficacy of Suby G in dissolving artificial brushite stones using an experimental arrangement simulating the physiological conditions in the upper urinary tract. Moreover, the efficacy of four different intracorporeal lithotripsy devices (electrohydraulic, pneumatic, laser and ultrasound) was tested and it was shown that electrohydraulic lithotripsy seems to be the best system for comminution of brushite stones. PMID- 10460898 TI - Duration of increased mucosal permeability of the urinary bladder after acute overdistension: an experimental study in rats. AB - The duration of damage to the mucosal barrier of the urinary bladder after overdistension was investigated in a rat model. Overdistension was induced for 3 h in 48 female Sprague-Dawley rats by forced diuresis and balloon obstruction of the bladder neck. In 24 rats 0.5 ml of 2% solution of Trypan blue in 0.9% NaCl solution was instilled into the bladder for 1 h at 0 h, 24 h, 48 h, 7 days and 21 days after overdistension. After dyeing, full-thickness samples were taken from the wall of the bladder dome and body immediately above the ureteral orifices for histological studies. Inflammatory reaction was investigated histologically without Trypan blue dyeing in 24 rats at 0 h, 24 h, 48 h, 7 days and 21 days after overdistension. At 0, 24 and 48 h after overdistension the bladder wall was deep blue throughout. The dome and body were similar. At 7 days there was only slight staining of the bladder surface urothelium and subjacent connective tissue, while at 21 days there was no longer any dye in the bladder wall or urothelium. Oedema reached its maximum at 48 h, and large numbers of inflammatory cells were seen in the submucosa at 48 h. These changes had normalized by 7 days. After overdistension urothelial integrity is destroyed for several days, making it possible for different substances in the urine to penetrate into the bladder wall. This renders questionable the use of bladder distension in the treatment of interstitial cystitis, as it may only increase leakage of the urothelium and accelerate inflammatory reaction in the bladder wall. However, in the present study of healthy rat bladders the integrity of the urothelium had recovered to a large extent after 1 week and completely after 3 weeks. PMID- 10460899 TI - Increased intraprostatic pressure in patients with chronic prostatitis. AB - The purpose of this prospective study was to develop a method for measuring intraprostatic pressure. Intraprostatic, extraprostatic and perineal subcutaneous pressures were measured in 43 patients. Twenty-four patients had chronic nonbacterial prostatitis (CNP) and prostatic hyperplasia (group A), 10 patients had benign prostatic hyperplasia (BPH) (group B) and 9 patients served as controls (group C). The pressure measurements were performed with a Stryker pressure monitor under transrectal ultrasonographic control at three different points: perineal subcutaneous tissue, paraprostatic tissue and the apex of the prostate beneath the capsule. Significantly higher intraprostatic pressure values (P < 0.001) were recorded in the patients with CNP compared with the BPH patients or the controls. We conclude that this novel method of measuring intraprostatic pressure, which has not been reported earlier, could be a new tool in the diagnosis of CNP and in the evaluation of the therapeutic effects of the different treatment modalities used in CNP. PMID- 10460900 TI - Evidence for only a moderate lipid peroxidation during ischemia-reperfusion of rat kidney due to its high antioxidative capacity. AB - The extent of lipid peroxidation after ischemia-reperfusion (I-R) injury in rat kidney has been controversial. After I, xanthine oxidase (XO) is thought to be the main oxygen radical-generating system and malondialdehyde (MDA) is considered to be a marker of lipid peroxidation (LPO). In young rats (10 weeks old) a unilateral warm I of 40 and 60 min duration with subsequent R up to 1 h was conducted. Beside the "footprints" of oxidative stress, the cytosolic antioxidative capacity, expressed as superoxide anion (SOA) scavenging capacity, and the renal catalase were also investigated. There was only a moderate and transient increase of renal MDA 5 and 10 min after the onset of reoxygenation (133.57/70. 67 and 97.84/91.57 vs. 49.47 nmol/g ww in preischemic controls). ATP breakdown (to 83/65 from 2947 nmol/g ww) with consecutive accumulation of hypoxanthine (up to 1105 nmol/g ww) at the end of ischemic period and the subsequent rapid decline of hypoxanthine by XO during reperfusion were used for an assessment of the SOA-generating capacity of these kidneys. Superoxide dismutase (SOD) activity, glutathione (GSH) and the high activity of catalase (18000 U/g ww) remained nearly unchanged during R. Only 1/25-1/50 of the kidney cytosol was able to scavenge the whole amount of SOA generated by the total XO activity of rat kidney. Thus, it could be analytically and stoichiometrically shown that after IR there is only a moderate oxidative stress in kidneys of young rats; this is due to their high SOA-scavenging capacity compared with their SOA generating ability. PMID- 10460901 TI - Does the immunocytochemical detection of epithelial cells in bone marrow (micrometastasis) influence the time to biochemical relapse after radical prostatectomy? AB - The detection of cytokeratin-positive bone marrow cells has been considered a prognostic factor in numerous malignant tumors. We investigated whether this was also valid for localized prostate cancer. Bone marrow aspirates were taken prior to radical prostatectomy from 169 consecutive patients with pT1/2 pN0 G1-3 adenocarcinoma of the prostate. The immunocytochemical detection of cytokeratin no. 18 (CK 18)-positive cells using monoclonal antibody CK 2 was interpreted as micrometastasis. Repeat marrow aspirations were performed at 6 months postoperatively and once a year thereafter. The patients were re-examined over a period of at least 10 and a maximum of 72 months (median 32 months). An increase in prostate specific antigen >/=0.5 ng/ml was considered a biochemical "relapse". One hundred and fifty-four patients had evaluable bone marrow aspirates, of which 74.7% were CK 18-negative and 25.3% positive. The latency period for biochemical relapse was 1481 days (median) in the CK 18-negative group and 1106 days (median) in the CK 18-positive group. This difference was not statistically significant. The CK 18-positive aspirates (n = 39) showed one positive cell in 20 cases, two positive cells in 8 and three or more positive cells in 11 cases. The preoperative number of cells had no statistically significant effect upon the onset of biochemical relapse. Only patients with three or more CK 18-positive cells tended to have a poorer prognosis. One hundred and thirteen patients had evaluable bone marrow aspirates pre- and postoperatively. Postoperative persistence or occurrence of CK 18-positive cells did not affect the outcome of the disease. The detection of CK 18-positive cells in bone marrow does not influence the prognosis of patients with localized prostate cancer within a period of 32 months (median). Solely a subgroup of patients showing a large preoperative number of CK 18-positive cells seems to tend to an unfavorable course of the disease. Thus, further studies are necessary aiming at a more detailed characterization of these cells. PMID- 10460902 TI - Circulating prostate-specific antigen mRNA during radical prostatectomy in patients with localized prostate cancer: with special reference to neoadjuvant hormonal therapy. AB - To determine the potential risk of hematogenous dissemination of prostate cancer cells during radical prostatectomy (RP), we investigated the pre- and intraoperative circulating prostate-specific antigen (PSA) mRNA in patients with clinically localized prostate cancer, with special reference to neoadjuvant hormonal therapy (NHT). Using a nested reverse transcriptase (RT) polymerase reaction (PCR) assay, PSA mRNA in the peripheral blood was evaluated pre- and postoperatively in a total of 23 patients, 10 of whom received NHT with antiandrogens. The RT-PCR assay employed detected one LNCaP cell in 10(7) mononuclear blood cells, and showed no positive signal in the blood samples from all 15 healthy controls. Pre- and intraoperative circulating PSA mRNA was positive in 11 (48%) and 18 patients (78%), respectively. All 11 patients with positive preoperative PSA mRNA continued to be positive during RP, and seven (58%) of 12 patients with negative preoperative PSA mRNA had a positive conversion. Although the patients' ages, preoperative serum PSA values and clinical or pathological stages were not associated with the pre- and intraoperative PSA mRNA results, the NHT group showed a significantly lower incidence of preoperative PSA mRNA positivity (2/10) than the group receiving RP alone (9/13) (20% vs 69%, P = 0.036). NHT, however, showed no suppressive effect on either intraoperative positivity or positive conversion of circulating PSA mRNA. The present study suggests that a substantial number of patients receiving RP are at risk of hematogenous dissemination, and NHT with antiandrogens has a minimal or no suppressive effect on the circulating PSA mRNA during surgical manipulation of the prostate. Because the clinical significance of circulating cancer cells remains to be determined, long-term follow-up in association with the circulating cancer cells assessed by the RT-PCR is essential in order to establish the role of molecular staging as well as NHT. PMID- 10460903 TI - Glutamine: a necessary nutrient for the intensive care patient. AB - Glutamine is a dispensable amino acid, a fact which is particularly important for intensive care patients, and it can be used as an oxidative substrate in processes which require prompt regulation of quantitatively large flows. The production and transport of glutamine from skeletal muscle may be inadequate in patients under intensive care, hence supplemented nutrition has been suggested. Improved long-term survival has been reported, which makes glutamine treatment one of the very few therapeutic strategies that improves outcome in intensive care. This overview deals with the metabolic and physiologic features and updates the clinical documentation of the field. PMID- 10460904 TI - Does glutamine reduce bacterial translocation? A study in two animal models with impaired gut barrier. AB - Failure of intestinal barrier function and subsequent translocation of bacteria from the gut are believed to play a decisive role in the development of systemic septic complications, for example, following major trauma or major abdominal surgery. This study evaluated: (a) the effect of glutamine on colonic microcirculation and electrophysiological parameters reflecting gut barrier function, (b) the translocation of live bacteria to extraintestinal organs, and (c) disease outcome in two animal models with impaired gut barrier function. Severe acute pancreatitis or colitis was induced in rats randomized for therapy with or without glutamine (0.5 g/kg daily). After 48 h one animal group was prepared for intravital microscopy of colonic capillary blood flow and electrophysiological measurement of gut permeability; another was killed after 96 h for histological and microbiological examination. In animals with pancreatitis, glutamine (Gln) supplementation significantly improved gut permeability, i.e., Gln increased colonic transmucosal resistance from 67+/-7 to 92+/-3 Omega/cm(2) and decreased mannitol flux through the epithelium by 53%. Capillary blood flow in the colonic mucosa was improved by 25%. The prevalence of pancreatic infections was reduced from 86% in animals on standard parenteral nutrition to 33% in animals given the Gln-enriched diet (P<0.05); mortality decreased by 32%. In colitis, Gln had no significant effect on these parameters except for improving colonic capillary blood flow in colon segments not adjacent to the major injury site. Glutamine supplementation improves colonic capillary blood flow, stabilizes gut permeability, and reduces secondary pancreatic infections and mortality in severe rodent pancreatitis, but it is not helpful in colitis. This confirms previous reports that glutamine stabilizes gut barrier function only in certain diseases. Our experimental data strongly suggest that acute pancreatitis (rather than colitis) is one of the diseases with gut barrier dysfunction in which glutamine substitution may be helpful to reduce bacterial translocation and should therefore be tested in a controlled clinical trial. PMID- 10460905 TI - Short-chain fatty acids stimulate mucosal cell proliferation in the closed human rectum after Hartmann's procedure. AB - Hartmann's procedure in humans results in a closed rectum deprived of its natural short-chain fatty acid source. This induces atrophy of the entire rectal wall. Ki 67 is a monoclonal antibody directed towards proteins in the cell nucleus that are present only during cell proliferation. This study investigated the effects of short-chain fatty acids on mucosal cell proliferation in the human rectum after Hartmann's procedure by means of Ki-67. Eight patients in whom Hartmann's procedure was performed were treated with placebo and then short-chain fatty acids for 14 days. Biopsies specimens were taken from the rectum before and after treatment; these were prepared with Ki-67 and labeling index was determined. The treatment was found significantly to increase proliferative activity in the rectal mucosa (P<0.01); the increase was principally in the middle (P<0.01) and upper crypt (P<0.05) compartments. PMID- 10460906 TI - Laparoscopic colectomy yields similar morbidity and disability regardless of patient age. AB - This study compared the outcome factors of morbidity and the length of disability in older and younger patients following laparoscopic colorectal surgery. All patients undergoing laparoscopic segmental resection during the study period were included. Morbidity was determined by reviewing the medical records, and disability by a patient-administered questionnaire. The series was divided into two age cohorts (/=65 years), which did not differ significantly in gender or type of procedure. Between these two groups we found no significant differences in mean duration of ileus (3.3 days in both groups), the mean length of hospitalization (5.7 vs. 6.3 days, respectively), morbidity rate (18% vs. 21%), or time until returning to partial activity (1.6 vs. 1.6 weeks) or to full activity (3 vs. 2 weeks). Our findings demonstrate that neither the morbidity rate nor the disability period after laparoscopic techniques differ between elderly and younger patients. We therefore endorse the use of laparoscopy regardless of patient age. PMID- 10460907 TI - A case-control-study comparing laparoscopic versus open surgery for rectosigmoidal and rectal cancer. AB - This study compared laparoscopic with open surgery for the cure of cancer of the rectosigmoid and rectum. Results of surgery, postoperative recovery, and oncological follow-up were compared between 32 laparoscopic curative procedures (19 laparoscopic-assisted anterior resections for cancer of the rectosigmoid or upper rectum and 13 laparoscopic abdominoperineal resections for low rectal cancer) and 32 controls matched for age, UICC stage, tumor site, and type of resection who underwent open surgery during the same observation period. Morbidity was identical after laparoscopic and open resection (31.3%). Surgery was equally radical in the two groups regarding yield of lymph nodes and lateral and distal margins. Survival, recurrence, and cancer-related mortality showed no statistical differences. There was no port-site recurrence. The benefits of laparoscopic surgery were shown with a reduction in perioperative blood transfusion and earlier return of bowel function. However, the operative time was significantly increased in the laparoscopic group. This study shows that laparoscopic surgery for the cure of colorectal cancer is technically feasible, and that oncological short-term outcome does not differ from the results achieved by open techniques. However, prospective randomized trials are mandatory to evaluate the definite role of laparoscopic surgery for malignancy. PMID- 10460908 TI - Total anorectal reconstruction by double graciloplasty: experience with delayed, selective use of implantable pulse generators. AB - This study reports our experience with total anorectal reconstruction (TAR), supported at a later phase, whenever necessary, by an implantable pulse generator. Thirteen patients underwent total anorectal reconstruction by double graciloplasty, diverting loop colostomy, and implantation of temporary electrodes. External-source, short-term, intermittent electrostimulation and biofeedback were used for neosphincter voluntary control training. After abdominal stoma closure, 6 months after initial surgery in disease-free patients, functional results were evaluated by a scoring system and anomanometry. A pulse generator was implanted whenever continence was judged unsatisfactory. After continuous electrostimulation training, neosphincter function was reassessed. Major graciloplasty complications (partial muscle necrosis and perineal colostomy necrosis) were treated successfully by surgery. One death of myocardial infarction occurred after discharge. Three patients refused further surgery. One patient did not undergo abdominal stoma closure because of early hepatic metastases. Functional evaluation after closure (eight patients) showed the following results: two "excellent" (no pulse generator implanted), three "good" (two stimulator implantations, with an "excellent" result), two "fair", and one "poor" (3 implantations, with a "good" result). In addition to improving clinical results (P=0.042), resting anal pressures were also increased significantly by active an implantable pulse generator (P=0.043). Although stimulators, whenever implanted, improved the neosphincter function, delayed, selective use of these in some cases rendered an implantable pulse generator either unnecessary from a functional viewpoint or redundant because of cancer recurrence or infectious complications. Drawbacks to the procedure were poor patient compliance to neosphincter training and to multiple surgical procedures, and excessive wasting of human resources during training for intermittent electrostimulation and biofeedback. PMID- 10460909 TI - Colonoscopy in octogenarians: a review of 428 cases. AB - Studies on the use of colonoscopy in the octogenarian are few. Therefore this study evaluated the results and cost-effectiveness of colonoscopy in octogenarians. A total of 403 patients 80 years of age or older who underwent colonoscopy from May 1994 to May 1996 were reviewed (median 84, range 80-95). Parameters evaluated were indications for colonoscopy, significant endoscopic findings (biopsy-confirmed adenocarcinoma and adenomatous polyps >/=1 cm), complications, colonoscopy completion rate, and mean charge per procedure. Postpolypectomy bleeding occurred in one patient. The cecal intubation rate was 94%. The calculated cost per procedure was U.S. $2,342. Indications for colonoscopy/number of cancers detected include: change in bowel habits, 78/2; blood/hemoccult positive, 69/8; abdominal pain, 12/0; constipation, 9/0; diarrhea, 8/0; surveillance for history of polyps, 159/3; surveillance for history of cancer, 51/1; cancer or polyp on sigmoidoscopy, 42/4. The cancer detection rate in patients with bleeding was 11.5%, compared with 1. 9% for all other symptoms. Colonoscopy can be safely performed in the octogenarian population. Our data suggest that more stringent selection criteria for colonoscopy in the octogenarian could result in significant cost savings. PMID- 10460910 TI - Palliative cryosurgery for rectal carcinoma. AB - Cryosurgery is one of the palliative treatment options to decrease local symptoms due to rectal carcinoma. A total of 106 patients (aged 45-92 years) underwent palliative cryosurgery for primary rectal cancer. Inclusion criteria were prohibitive surgical risk, unresectability, distant metastases, patient refusal of surgical intervention, and old age. The palliative effect of cryosurgery on local symptoms was classified as good, moderate, or none. The duration of palliation was calculated in relation to survival as a palliative index: the number of months of (moderate or good) palliation divided by the number of months of survival from the start of the treatment, multiplied by 100. In 66 patients (62%) there was complete relief of local symptoms, with a palliative index of 88%. In 17 patients (16%) palliation was moderate, and in 23 patients (22%) no palliation was achieved. The 50% survival duration in the 66 patients was 33 months. Especially symptoms of blood and/or mucous discharge were easily alleviated. Good alleviation of local symptoms was associated with the extent of the primary tumor process. Palliative effect was be assessed after a few treatments. Patients with circular rectal tumors seldom had long-term palliative effects of cryosurgery. Our findings show that Cryosurgery is a simple and safe treatment for rectal cancer. It should be considered for alleviation of local symptoms in patients with rectal cancer who are unsuitable for radical surgery. PMID- 10460913 TI - Instability, stabilization, and formulation of liquid protein pharmaceuticals. AB - One of the most challenging tasks in the development of protein pharmaceuticals is to deal with physical and chemical instabilities of proteins. Protein instability is one of the major reasons why protein pharmaceuticals are administered traditionally through injection rather than taken orally like most small chemical drugs. Protein pharmaceuticals usually have to be stored under cold conditions or freeze-dried to achieve an acceptable shelf life. To understand and maximize the stability of protein pharmaceuticals or any other usable proteins such as catalytic enzymes, many studies have been conducted, especially in the past two decades. These studies have covered many areas such as protein folding and unfolding/denaturation, mechanisms of chemical and physical instabilities of proteins, and various means of stabilizing proteins in aqueous or solid state and under various processing conditions such as freeze-thawing and drying. This article reviews these investigations and achievements in recent years and discusses the basic behavior of proteins, their instabilities, and stabilization in aqueous state in relation to the development of liquid protein pharmaceuticals. PMID- 10460914 TI - Concomitant and controlled release of dexamethasone and 5-fluorouracil from poly(ortho ester). AB - A viscous bioerodible and hydrophobic poly(ortho ester) has been developed as a biocompatible, sustained drug release system for an ophthalmic application in intraocular proliferative disorders. The combination of wound healing modulators such as 5-fluorouracil and dexamethasone is a major advantage since these drugs act at different stages of these diseases. Since 5-fluorouracil is an acidic, water-soluble compound and dexamethasone exists in three chemical forms, i.e. the water-insoluble base, the highly hydrophobic acetate ester or the basic phosphate salt, it was of interest to investigate whether the physicochemical properties of the drugs have an influence on their release rates, and whether a concomitant and sustained release of both 5-fluorouracil and dexamethasone could be achieved. It has been found that lipophilicity and acidobasicity play a major role in controlling drug release rates and polymer degradation. The combination of 5 fluorouracil and dexamethasone phosphate allows a sustained and concomitant release of both drugs, due to the basic characteristics of the corticosteroid which stabilize the polymer. This system appears to be promising for concomitant and controlled drug delivery aimed at the pharmacological treatment of intraocular proliferative disorders. PMID- 10460915 TI - Effect of formulation on the systemic absorption of insulin from enhancer-free ocular devices. AB - Several Gelfoam (absorbable gelatin sponge, USP) based surfactant free devices containing either sodium or zinc insulin were prepared with diluted acetic or hydrochloric acid. They were evaluated by the lowering of the blood glucose concentration in rabbits. The systemic absorption of insulin from the device can be enhanced by using a 5% or higher concentration of acetic acid solution as well as 1% HCl solution. The results indicate that the proposed device prepared with up to 30% of acetic acid solution produced no eye irritation. A single device containing 0.2 mg of insulin is sufficient to control the blood glucose levels in a uniform manner (60% of initial) for over 8 h. PMID- 10460916 TI - Ultrasound-enhanced diffusion into coupling gel during phonophoresis of 5 fluorouracil. AB - PURPOSE: To investigate the competitive transport across skin and back-diffusion of 5-fluorouracil into coupling gel under the influence of ultrasound, heat-alone and Azone enhancement. METHODS: The ultrasound effect on 5-fluorouracil penetration through whole rat skin was investigated in modified diffusion cells using a commercial ultrasound generator which was calibrated with a bilaminar membrane hydrophone. RESULTS: Ultrasonic dosimetry measurements demonstrated that the skin membrane was subjected to a complex and unpredictable standing wave field which induced physiologically acceptable heating of the tissue. Surprisingly, ultrasonication produced a decrease in percutaneous drug penetration. Quantification studies indicated that this effect was due to the diffusive loss of the hydrophilic substance 5-fluorouracil from the skin surface into the overlying volume of coupling gel. This phenomenon could be duplicated by the application of conductive heating, indicating that the thermal effects of ultrasound were probably responsible for accelerated 5-fluorouracil diffusion through the gel. CONCLUSION: This study acutely demonstrates how formulation design of the donor vehicle/coupling gel may radically affect therapeutic efficacy in phonophoretic systems. PMID- 10460917 TI - TR146 cells grown on filters as a model of human buccal epithelium: III. Permeability enhancement by different pH values, different osmolality values, and bile salts. AB - The aim of the present study was to evaluate the TR146 cell culture model as an in vitro model of human buccal epithelium with respect to the permeability enhancement by different pH values, different osmolality values or bile salts. For this purpose, the increase in the apparent permeability (P(app)) of the hydrophilic marker mannitol due to exposure to solutions with pH values or osmolality values different from the physiological values was studied. As in studies with solutions of either taurocholate (TC), glycocholate (GC) or glycodeoxycholate (GDC) the results were compared to the increase in P(app) of mannitol obtained in analog studies using porcine buccal mucosa in an Ussing chamber. The effect of the exposure on the electrical resistance of the TR146 cell culture model and the porcine buccal mucosa was measured, and the degree of protein leakage due to GC exposure was investigated in the TR146 cell culture model. The porcine buccal mucosa was approximately ten times less permeable to mannitol than the TR146 cell culture model. The P(app)TC. Increased P(app) values correlated with a decrease in the electrical resistance of the TR146 cell culture model and the porcine buccal mucosa. GC was shown to induce concentration dependent protein leakage in the TR146 cell culture but only from the site of application, and the results indicate that duration of exposure further than 120 min was of minor importance. The present results indicate that the TR146 cell culture model may be a suitable in vitro model for efficacy studies and mechanistic studies of enhancers with potential use in human buccal drug delivery. PMID- 10460918 TI - Degradation study of the investigational anticancer drug clanfenur. AB - Clanfenur belongs to a new group of substituted benzoylphenylureas. The drug shows both in vitro and in vivo antitumour activity. To assess its chemical stability, a study was carried out in which the effect of pH, temperature, ionic strength and buffer concentration on the reaction rate constant k(obs) were examined. A stability-indicating reversed-phase high performance liquid chromatography (RP-HPLC) system was used. The pH-log k(obs) degradation profile, obtained at 70 degrees C, shows that clanfenur has its maximum stability in the pH region 4-5. At pH 7, half-lives were calculated by extrapolation of the Arrhenius plot; at 4 degrees C the half-life was calculated to be 141 years and at 25 degrees C 9. 5 years. The activation energy was calculated to be 114 kJ/mol. In acidic, neutral, and alkaline media, the ionic strength has no effect on the degradation. The buffer concentration of citrate, phosphate, borate, and carbonate did not affect the value of k(obs). An RP-HPLC chromatogram of degraded clanfenur shows the presence of four degradation products, three of which were identified by LC-ESI-MS as p-chloroaniline, p-chlorophenylurea and 2-fluoro-6 dimethylaminobenzamide. PMID- 10460919 TI - Powder filling into hard gelatine capsules on a tamp filling machine. AB - A series of pharmaceutical excipient powders used in capsule formulations as fillers have been filled into hard gelatine capsules on a Bosch GKF-400S tamp filling machine. These machines depend on pushing pins through a powder bed so that a unit dose is transferred into a dosing disc. This dose is then ejected into the capsule body. The results indicate that the range of powders that can be filled on this type of machine exceeds that applicable to a dosator nozzle system. Filling problems due to powder flooding could be solved by increasing the powder bed height in the powder bowl. The fact that such powders usually do not form a firm plug was not reflected in the coefficient of fill weight variation, and uniform filling could hence be achieved without problems. The influence of the powder bed height on the capsule fill weight increased with decreasing powder flow. The influence of the setting of the tamping pins on the capsule fill weight was comparatively small and further decreased with a decrease in powder flow. However, when a granulated product (Elcema G250) was filled, the tamping pin setting was more important than the influence of the powder bed height on the capsule fill weight. For moderate flowing powders the coefficient of fill weight variation appeared to be nearly independent of powder bed height or tamping pin setting. However, the filling performance of powders with poor flow properties could be adjusted by optimising both machine settings. Very complex relationships were found between the powder properties such as angle of internal flow, dynamic densification profile, Carr's compressibility index and particle size and shape, and the filling behaviour. PMID- 10460921 TI - Correction: Cisplatin, Radiation, and Adjuvant Hysterectomy Compared with Radiation and Adjuvant Hysterectomy for Bulky Stage IB Cervical Carcinoma. PMID- 10460920 TI - Formulation strategies for the stabilization of tetanus toxoid in poly(lactide-co glycolide) microspheres. AB - The development of a single-dose tetanus vaccine based on Poly(Lactic acid) (PLA) or Poly(Lactide-co-Glycolide) (PLGA) microspheres has been complicated due to the instability of tetanus toxoid (TT) inside these systems. Herein we report an attempt to re-design PLGA microspheres by co-encapsulating TT in the dry solid state together with potential protein stabilizers, such as trehalose, bovine serum albumin, alginate, heparin, dextran or poloxamer 188 and by using an appropriate microencapsulation technique. These newly developed PLGA microspheres were able to release in vitro antigenically active TT for at least 5 weeks, the amount released being highly dependent on the stabilizing excipient used. More specifically, results showed that dextran and heparin provided a particularly stabilizing environment for TT inside the microspheres during the polymer degradation process. The efficacy of this strategy was demonstrated by the high, long lasting titers of neutralizing antibodies achieved after in vivo administration of dextran-containing microspheres with a small amount of alum adsorbed TT, as compared to the commercial adsorbable tetanus vaccine. These findings suggest that future developments in the area of vaccinology depend on ability to combine a detailed knowledge of the microencapsulation technology with a rational choice of stabilizing excipient or combination of excipients. PMID- 10460922 TI - Biomass cooking fuels and prevalence of tuberculosis in India. AB - OBJECTIVES: To examine the relation between use of biomass cooking fuels (wood or dung) and prevalence of active tuberculosis in India. METHODS: The analysis is based on 260,162 persons age 20 and over in India's 1992-93 National Family Health Survey. Logistic regression is used to estimate the effects of biomass fuel use on prevalence of active tuberculosis, as reported by household heads, after controlling for a number of potentially confounding variables. RESULTS: Persons living in households that primarily use biomass for cooking fuel have substantially higher prevalence of active tuberculosis than persons living in households that use cleaner fuels (odds ratio [OR] = 3.56; 95% confidence interval [CI] = 2.82-4. 50). This effect is reduced somewhat when availability of a separate kitchen, house type, indoor crowding, age, gender, urban or rural residence, education, religion, caste or tribe, and geographic region are statistically controlled (OR = 2.58; 95% CI = 1.98-3.37). Fuel type also has a large effect when the analysis is done separately for men (OR = 2.46; 95% CI = 1.79-3.39) and women (OR = 2. 74; 95% CI = 1.86-4.05) and separately for urban areas (OR = 2.29; 95% CI = 1.61-3.23) and rural areas (OR = 2.65; 95% CI = 1.74 4.03). The analysis also indicates that, among persons age 20 years and over, 51% of the prevalence of active tuberculosis is attributable to cooking smoke. CONCLUSIONS: Results strongly suggest that use of biomass fuels for cooking substantially increases the risk of tuberculosis in India. PMID- 10460923 TI - Fatal dengue hemorrhagic fever in Cuba, 1997. AB - OBJECTIVES: After more than 15 years without dengue activity, a dengue II epidemic was reported in Cuba in 1997. Three thousand and twelve serologically confirmed cases were reported, with 205 dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) cases and 12 fatalities. This report presents the clinical, serologic, and virologic findings in the 12 fatal DHF/DSS cases. METHODS: Serum and necropsy samples were studied by viral isolation in C636 cell line and polymerase chain reaction. Serum samples were tested by IgM capture enzyme-linked immunoassay (ELISA) and ELISA inhibition method (EIM). RESULTS: All 12 cases were classified as DHF/DSS according to the Pan American Health Organization Guidelines for Control and Prevention of Dengue and Dengue Hemorrhagic Fever in the Americas. All patients were older than 15 years. Women were more frequently affected. The symptoms and signs presented by these patients were similar to those previously described in DHF/DSS cases. Clinical deterioration occurred on average at day 3.75. Abdominal pain and persistent vomiting were the earliest and most frequent warning signs. Dengue infection was confirmed in all cases. IgM antibodies were detected in 11 of 12 cases, all of them with a secondary infection. Dengue II virus was detected by viral isolation in 12 samples and by polymerase chain reaction in 17. Virus or RNA was detected in various tissues, including kidney, heart, lung, and brain. CONCLUSION: The clinical, pathologic, and laboratory features of 12 cases of fatal dengue hemorrhagic fever were reviewed. The results obtained demonstrate that adults with a primary dengue infection are at risk of developing the severe disease (DHF) if they are infected with a different serotype. PMID- 10460924 TI - Imported malaria in a Singapore hospital: clinical presentation and outcome. AB - OBJECTIVE: To evaluate the clinical presentation and outcome of imported malaria. METHODS: A retrospective chart review was conducted of patients with imported malaria admitted to the Communicable Disease Centre (CDC), Singapore (a 130-bed tertiary referral center) from January 1992 to December 1993. An imported case was defined as a smear-positive infection that was acquired in another country. RESULTS: Among 200 malaria patients hospitalized at CDC, 168 imported cases (137 males and 31 females, 131 nonresidents and 37 residents) were studied. The mean age was 31.6 6 10.5 years. The countries visited were India (49.4%), Indonesia (16.7%), and Bangladesh (13%). Five patients had chemoprophylaxis and 36 patients had experienced previous malaria infection. The predominant symptoms were fever (97.6%), chills (79.2%), and rigors (67.9%). Hepatomegaly was detected in 56 (33.3%) and splenomegaly in 49 patients (29.2%). Plasmodium vivax was present in 132 patients, Plasmodium falciparum in 29, and mixed P. vivax and P. falciparum in 7 patients. Parasitemia ranged from 0.1% to 8.0%. Of the vivax cases, 130 were treated with chloroquine, followed by primaquine in 123 patients. Quinine was given to 36 patients (29 falciparum malaria and 7 mixed infections). Median time to fever defervescence was 2 days. Complications occurred in three patients (2 with shock and 1 with pulmonary edema). According to World Health Organization gravity criteria, body temperature over 40 degrees C was detected in six patients, bilirubinemia higher than 50 mmol/L in nine, parasitemia over 5% in five, glycemia less than 2.2 mmol/L in two patients. There were five relapses. No death was recorded. CONCLUSION: Plasmodium vivax is the most common cause of imported malaria, with the majority acquired from the Indian subcontinent. Only a few patients presented with severe malaria. PMID- 10460925 TI - Parapertussis and pertussis: differences and similarities in incidence, clinical course, and antibody responses. AB - OBJECTIVES: To compare the incidence, clinical course, and serologic response to Bordetella antigens in patients with parapertussis and pertussis. DESIGN: Two studies were performed in Sweden during the 1990s, when pertussis vaccines were used only in clinical trials. Study I was a retrospective study of patients with positive Bordetella cultures obtained in clinical routine, and study II involved an active search for patients with Bordetella infections during a placebo controlled trial of a pertussis toxoid vaccine. RESULTS: Study I includes 58, and study II 23 patients with parapertussis. In study I, the incidence of parapertussis was 0.016 cases per 100 person years in children 0 to 6 years old and 0 in older children and adults. In study II, the incidence rates of parapertussis and pertussis were 0.2 and 16.2 per 100 person years, respectively, in children followed from 3 months to 3 years of age. The median number of days with cough was 21 in parapertussis and 59 in pertussis. The proportions of children with whooping and vomiting were lower in parapertussis than in pertussis. Geometric mean serum filamentous hemagglutinin IgG increased from 6 to 63, and pertactin IgG from 4 to 12 units/mL in parapertussis patients, which was similar to increases in children with pertussis. CONCLUSIONS: Disease caused by Bordetella parapertussis is diagnosed less commonly and is milder and of shorter duration than disease caused by Bordetella pertussis. Parapertussis induced serum IgG against filamentous hemagglutinin and pertactin of similar magnitude as does pertussis, and did not induce serum IgG against pertussis toxin. PMID- 10460926 TI - Severe pneumococcal infection at a Thai hospital. AB - OBJECTIVES: To determine clinical manifestations, prognostic factors, and therapeutic outcomes of severe pneumococcal infection. METHODS: Hospitalized patients with specimens cultured positive for Streptococcus pneumoniae were identified retrospectively by reviewing hospital records from 1992 to May 1998 at Siriraj Hospital. RESULTS: Of 205 evaluable cases, 130 (63.4%) patients were male. Nineteen (9.3%) patients were less than 2 years old, 29 (14.1%) were between 2 and 13 years, 99 (48.3%) were between 14 and 60 years, and 58 (28.3%) were over 60 years of age. From 1992 to 1997, the average admission rate was highest (36.4%) between January and March (range = 20-45%). Average admission rates during other periods ranged from 20.0% to 23.1%. Pneumonia (50.7%) and acute exacerbation of chronic obstructive pulmonary disease or infected bronchiectasis or bronchopneumonia (21.0%) were the most frequent diagnoses, followed by meningitis (14.6%) and primary sepsis without localized lesion (8. 3%). The mortality rate during the first 7 days of hospitalization was 28.8%, and thereafter, 11.7%. The odds ratios (95% CI) of old age, congestive heart failure, and alcoholism for death were 3.4 (1. 4-8.2), 8.6 (0.97-76.1), and 8.0 (3.1 20.9), respectively. For pneumonitis only, mortality rates among alcoholic and nonalcoholic patients were 76.9% and 39.6%, respectively (P = 0.025). CONCLUSIONS: Patients who were alcoholic, over 60 years of age, or had congestive heart failure were vulnerable to severe pneumococcal infection with significant mortality, in spite of proper selection of empirical antimicrobials. Diabetes mellitus and multiple myeloma also contributed to late mortality after 7 days of hospitalization. PMID- 10460927 TI - Antimicrobial resistance among community-acquired pneumonia isolates in Europe: first results from the SENTRY antimicrobial surveillance program 1997. SENTRY Participants Group. AB - OBJECTIVE: The SENTRY antimicrobial surveillance program was established to monitor the occurrence and antimicrobial susceptibility of bacterial pathogens via an international network of sentinel hospitals. MATERIAL AND METHODS: Microorganisms were forwarded to the reference laboratory for testing against various antimicrobial agents using broth microdilution. Twenty European hospitals referred 286 Streptococcus pneumoniae, 309 Haemophilus influenzae, and 167 Moraxella catarrhalis isolates during the first 10 months of the study, starting in April 1997. RESULTS: Seven percent of the S. pneumoniae isolates were highly resistant to penicillin, and 21% showed intermediate resistance. The highly resistant pneumococcal isolates came from Coimbra, Barcelona, Athens, and London, whereas the intermediate penicillin-resistant isolates were received from all participating countries. The incidence of intermediate penicillin-resistant pneumococci was lowest in Lausanne, Freiburg and Duesseldorf, London, and Utrecht and highest in southern European countries. Fifty-five percent of the penicillin resistant S. pneumoniae were also resistant to erythromycin, and 35% to clindamycin. Sparfloxacin, trovafloxacin, levofloxacin, and vancomycin were fully active against pneumococcal isolates. Haemophilus influenzae isolates were generally highly susceptible to most of the antibiotics tested, and 92% of the M. catarrhalis isolates were resistant to penicillin. Susceptibility to cephalosporins, ciprofloxacin, levofloxacin, and rifampicin was 100%. CONCLUSION: Penicillin may no longer be the first-choice drug for empirical treatment of pneumococcal infections. The newer fluoroquinolones may play a role in the empirical treatment of community-acquired pneumonia. PMID- 10460929 TI - Angiostrongylus cantonensis eosinophilic meningitis. AB - In the past 50 years, Angiostrongylus cantonensis, the most common cause of eosinophilic meningitis, has spread from Southeast Asia to the South Pacific, Africa, India, the Caribbean, and recently, to Australia and North America, mainly carried by cargo ship rats. Humans are accidental, "dead-end" hosts infected by eating larvae from snails, slugs, or contaminated, uncooked vegetables. These larvae migrate to the brain, spinal cord, and nerve roots, causing eosinophilia in both spinal fluid and peripheral blood. Infected patients present with severe headache, vomiting, paresthesias, weakness, and occasionally visual disturbances and extraocular muscular paralysis. Most patients have a full recovery; however, heavy infections can lead to chronic, disabling disease and even death. There is no proven treatment for this disease. In the authors' experience, corticosteroids have been helpful in severe cases to relieve intracranial pressure as well as neurologic symptoms due to inflammatory responses to migrating and eventually dying worms. PMID- 10460928 TI - Amphotericin B with and without itraconazole for invasive aspergillosis: A three year retrospective study. AB - BACKGROUND: Treatment of invasive aspergillosis is frequently unsuccessful, so innovations in therapy are needed. Clinical studies demonstrate that itraconazole may be an effective alternative to amphotericin B. Itraconazole also has been combined with amphotericin B in animal models of aspergillosis, but this regimen produced antagonistic effects. OBJECTIVES: To determine the role of itraconazole in the adjunctive treatment of invasive aspergillosis. METHODS: A review was conducted of all patients with definite or probable aspergillosis from January 1995 to December 1997 who were treated with conventional amphotericin B alone or in combination with itraconazole. RESULTS: Of 21 patients, 10 received amphotericin B and 11 received the combination. The two groups of patients were comparable clinically at baseline (including similar mean APACHE III scores). Both groups received similar doses and days of amphotericin B treatment. Of the patients who received combination therapy, nine (82%) were cured or improved, and of those who received only amphotericin B, five (50%) were cured or improved. CONCLUSIONS: This study demonstrates that itraconazole and amphotericin B given together are not clinically antagonistic and that the promise of combination therapy for aspergillosis should be evaluated further in a randomized clinical trial. PMID- 10460930 TI - Epidemiology and management of diarrheal disease in HIV-infected patients. AB - Diarrhea is the most common gastrointestinal symptom in human immunodeficiency virus (HIV) infection. It affects up to 90% of patients, becoming more frequent and severe as the immune system deteriorates. It often is associated with significant morbidity and mortality particularly in the developing countries. Gastrointestinal infections, some of which are attributable to inadequate sanitation and poor hygiene are the predominant cause of diarrhea, although multiple medications, including traditional herbs, also may be causes. The basic principles of management include detection of treatable causes, relief of symptoms, prevention of malnutrition, and psychosocial support. In up to 60% of cases, no cause can be identified, partly because of inadequate investigative facilities. Symptomatic treatment is the mainstay of management particularly when no cause can be identified. Unfortunately this can be extremely difficult when the patient is severely immune-suppressed. There is poor response to motility control drugs, such as loperamide, and others, such as octreotide, are too expensive. Fluid replacement should be started early to prevent excessive dehydration. This should be combined with nutritional support to prevent malnutrition. Psychosocial support, including counselling, for both the patient and the caring relatives, is required to alleviate anxiety, particularly when the diarrhea becomes intractable. PMID- 10460931 TI - On the etiology of tropical spastic paraparesis and human T-cell lymphotropic virus-I-associated myelopathy. AB - The purpose of this review is to present some concepts on the etiology of tropical spastic paraparesis or human T-cell lymphotropic virus-I (HTLV-I) associated myelopathy (TSP/HAM). The large number of syndromes that have been associated with HTLV-I (60 to date), the existence of TSP/HAM cases associated with other retroviruses (human immunodeficiency virus-2 [HIV-2], HTLV-II), the existence of many TSPs without HTLV-I, and the evidence of clear epidemiologic contradictions in TSP/HAM indicate that the etiopathogenesis of TSP/HAM is not yet clear. Tropical spastic paraparesis/HAM affects patients of all human ethnic groups, but usually in well localized and relatively isolated geographic regions where HTLV-I has been endemic for a long time. Environmental factors and geographic locations appear to be critical factors. Because the neuropathology of TSP/HAM suggests a toxometabolic, rather than a viral cause, it is proposed that an intoxication similar to neurolathyrism could account for some of TSP/HAM cases, mainly in tropical and subtropical countries. If this were the case, HTLV I could be a cofactor or act as a bystander. it is possible that co-infection with another agent is necessary to produce TSP/HAM and most of the syndromes associated with HTLV-I. PMID- 10460932 TI - Diuretic-induced hyponatremia. AB - Diuretics are one of the most common causes of severe hyponatremia. Yet, despite several relevant studies and years of clinical experience, the mechanism and optimal treatment of diuretic-induced hyponatremia remain unclear. What is clear is that most cases are caused by thiazide rather than loop diuretics and that severe hyponatremia can develop very rapidly in susceptible patients. In this review, I will discuss the pathogenesis, clinical features, prevention, and treatment of diuretic-induced hyponatremia in the hope that increased awareness and understanding will reduce the incidence and complications of this potentially life-threatening syndrome. PMID- 10460933 TI - IgA-antigliadin antibodies in patients with IgA nephropathy: the secondary phenomenon? AB - Circulating IgA-antigliadin antibodies (IgA-AGA) are often found in patients with IgA nephropathy (NP). IgA-AGA are sensitive markers of an abnormal immune system reaction to gluten, seen particularly in patients with celiac disease. However, a lack of IgA-antireticulin and IgA-antiendomysium antibodies and often jejunal mucosal atrophy of patients with IgA NP suggest that most patients do not have latent celiac disease. To examine the relationship between IgA-AGA and clinical data, enzyme-linked immunosorbent assays for IgA-AGA were performed in 28 patients with IgA NP and in 50 healthy persons. The results were calculated in arbitrary units (AU). The cutoff level for a negative or a positive test was found to be 60 AU, calculated according to the AGA test result (mean + 3 SD) in 50 healthy persons. The following clinical data were assessed: age, gender, disease duration, daily proteinuria, blood pressure, serum creatinine, and creatinine clearance. Control sera were negative for IgA-AGA. Positive IgA-AGA tests were observed in 14 of the 28 patients (p < 0.0001 vs. controls) and high levels of IgA-AGA (AU >90) in 6 of the 28 patients (p < 0.001 vs. controls). The mean duration of the disease of the patients with positive IgA-AGA was significantly longer as compared with the patients who had a negative antibody test. IgA-AGA correlated with age (p < 0.05, r = 0. 56), disease duration (p < 0.05, r = 0.40), and blood pressure (p < 0.05, r = 0.48). Antireticulin and antiendomysium antibody tests were negative in all patient and control sera. We conclude that IgA-AGA are associated with the progression of IgA NP. Our findings support the current concept about the pathogenesis of IgA NP, where the defective IgA production itself may be the primary and intestinal lesions as well as the production of IgA-AGA the secondary phenomenon. PMID- 10460934 TI - Intradialytic dobutamine therapy in maintenance hemodialysis patients with persistent hypotension. AB - Intradialysis hypotension is a common problem, especially in patients with poor left-ventricular function. We studied 6 patients who were on maintenance hemodialysis with left-ventricular ejection fraction of <40%, whose dialysis sessions were often complicated with severe hypotension (systolic blood pressure <90 mm Hg). Dobutamine infusion during dialysis significantly reduced the number of hypotensive episodes, increased left-ventricular ejection fraction, and decreased the number of emergency admissions to the hospital. PMID- 10460935 TI - Evidence for genetic heterogeneity in benign familial hematuria. AB - Benign familial hematuria (BFH: MIM141200) is an autosomal-dominant disease accounting for one-fifth of all hematuria of unknown cause in children. Previous observations suggest that BFH may be allelic to recessive Alport syndrome (AS: MIM 203780) with a mutation in the COL4A3/COL4A4 locus. However, it is not clear whether all cases of BFH are due to heterozygous mutation of COL4A3/COL4A4 genes. We report here the exclusion of linkage between BFH and COL4A3/COL4A4 loci at 2q35-37 in a restricted population from Sicily (Italy). Total lod score is -9.6 at theta 0. Furthermore, in some cases exclusion of linkage is evident even considering single families. We conclude that BFH is genetically heterogeneous. PMID- 10460936 TI - Clinical findings and outcomes of intra-hemodialysis cardiopulmonary resuscitation. AB - Renal failure with severe uremia is still an important cause of mortality, despite effective renal replacement therapy. Cardiopulmonary arrest (CPA) is the most severe complication during hemodialysis (HD). To acquire more information about cardiopulmonary resuscitation (CPR) during HD, we retrospectively enrolled 24 patients (11 males and 13 females) who had CPR during HD in a medical center during a 3-year period. Their mean age was 66.8 +/- 16.8 years. The CPR rate of the patients from our outpatient department (0.02%) was significantly lower than that from general wards (0.11%), the intensive care unit (ICU, 0.16%), or the emergency room (ER, 0.38%). Eighteen patients (75%) were initially resuscitated successfully. Only 11 patients (45.8%) survived more than 24 h after CPR, and 2 patients (8.3%) survived more than 1 month, but none survived until discharge. The rates of surviving 24 h and surviving to discharge during HD were lower than those in the general wards, the ICU or the ER. Sepsis (33.3%) and cardiogenic shock (25%) were the two leading causes of death. For analyzing factors affecting the outcome of CPR, we divided the patients into 2 groups by survival time (<==24 vs. >24 h). Patients with heart disease or with prolonged CPR durations (>30 min) had shorter survival. No significant survival difference between the 2 groups was found due to factors of age, sex, diabetic nephropathy, pre-arrest morbidity scores, pre-arrest laboratory data, renal failure pattern, HD duration, the preceding HD time and ultrafiltrated volume. PMID- 10460937 TI - Relapsing membranous nephropathy. Response to therapy of relapses compared to that of the original disease. AB - BACKGROUND: Although controversial, treatment of membranous nephropathy appears to yield a reduction in the degree of proteinuria and conservation of renal function. METHODS: We report herein our experience with the treatment with steroids alone (group II, n = 13), or in combination with immunosuppressants (group III, n = 19) of patients with membranous nephropathy and the nephrotic syndrome, with a mean follow-up of 8.37 years. RESULTS: All patients underwent a first remission, with 24-hour urine protein excretion falling to 0.63 +/- 0.25 g in group II and 0.62 +/- 0.26 g in group III (p = NS) after 12.69 +/- 10.94 months of treatment in group II and 18.95 +/- 13.17 months in group III (p = NS). Three patients from group II (23%) and seven patients from group III (36.8%) experienced four and eight relapses, respectively (proteinuria in 24 h 4.0 +/- 0.80 g in group II relapsers and 4.4 +/- 0.87 in group III relapsers; p = NS). On treatment, all relapses remitted (second remission) after 7 +/- 6.93 months of therapy for group II and 8.6 +/- 6.70 months of treatment for group III (p = NS). Thereafter, no patients from group II, but 3 patients from group III (33.3%) had a second relapse. After treatment, all relapses remitted (third remission) in 3.3 +/- 1.53 months of therapy. CONCLUSIONS: These studies show that relapses, which occur in one-third of patients, respond favorably to treatment albeit remitting in approximately half the time, and that the duration of remission gets progressively longer in the later compared to the earlier remission. PMID- 10460938 TI - Improvement of myocardial fatty acid metabolism through L-carnitine administration to chronic hemodialysis patients. AB - The concentration of carnitine, which is essential to fatty acid metabolism, can decrease markedly in patients on long-term hemodialysis coincident with life threatening cardiac damage. However, administration of L-carnitine improves the myocardial function of these patients. To evaluate the underlying events of this phenomenon, we used recently developed technology, (123)I-labeled beta-methyl-p iodophenyl-pentadecanoic acid (BMIPP) myocardial scintigraphy, as a test of myocardial fatty acid metabolism. Our results showed that the free carnitine concentration (19.2 +/- 6.5 micromol/l) was lower in 11 chronically dialyzed patients than in 8 healthy controls (49.3 +/- 7.7 micromol/l, p < 0. 0001). Additionally the heart to mediastinal ratio (H/M) of BMIPP was higher for these patients than for the controls (1.91 +/- 0.19 vs. 1.52 +/- 0.24, p < 0.005), and the patients' washout rate (WOR) of BMIPP was lower (17.2 +/- 6.0 vs. 22.8 +/- 4.2%, p < 0.05). After L-carnitine was administered orally to the patients at doses of 1 g/day for 1 month and 0.5 g/day for the following month, the concentration of free carnitine in their sera increased to 85.4 +/- 27.0 micromol/l (p < 0.0001). Although the H/M ratio did not change (1.89 +/- 0.20) with this treatment, their WOR increased to 21.9 +/- 6.6% (p < 0.001), similar to that of controls. The left ventricular end-diastolic dimension and left ventricular fractional shortening remained unchanged, as shown by echocardiography. The results presented here denote that a carnitine deficiency in chronically hemodialyzed patients disrupts their myocardial fatty acid metabolism, which is improved by L-carnitine supplementation. PMID- 10460939 TI - The plasma leptin concentration is closely associated with the body fat mass in nondiabetic uremic patients. AB - Plasma leptin is associated with the body mass index and, more precisely, with the body fat mass. Plasma leptin has been found to be elevated in uremic patients. This study aimed at investigating the plasma leptin concentration and associations between plasma leptin, body fat mass, and glomerular filtration rate in nondiabetic predialysis uremic patients and in nondiabetic patients on chronic hemodialysis. Plasma leptin, body fat mass, and creatinine clearance were measured in 22 predialysis uremic patients, 18 hemodialysis patients, and 24 healthy control subjects. The logarithmically transformed plasma leptin concentration was closely associated with the body fat mass in all groups (r = 0.93, r = 0.83, and r = 0.72, respectively; p < 0.000001, < 0.000002 and p < 0.001, respectively). In predialysis uremic patients the plasma leptin concentration was slightly elevated as compared with controls 10.4 (3.1-59.5) ng/ml versus 5.4 (1.6-47.5) ng/ml (median and range in parentheses; p < 0. 05), whereas the plasma leptin concentration was normal in hemodialysis patients. Plasma leptin was not significantly associated with the creatinine clearance in predialysis patients. In conclusion; the glomerular filtration rate seemed to have a limited influence on the plasma leptin concentration in nondiabetic uremic subjects matched by body fat mass to controls. The plasma leptin concentration was closely associated with the body fat mass, and the leptin level might, therefore, be useful as an indicator of the fat mass in nondiabetic uremic patients. PMID- 10460940 TI - Efficiency of 1-year treatment with fluvastatin in hyperlipidemic patients with nephrotic syndrome. AB - The influence of fluvastatin, a liver-selective, competitive inhibitor of the 3 hydroxymethyl-glutaryl-coenzyme A reductase, on the lipoprotein metabolism was investigated in 9 patients with nephrotic syndrome. All patients had biopsy proven renal disease as cause of their nephrotic syndrome and exhibited severe hyperlipidemia [baseline: serum cholesterol 358 +/- 46 mg/dl (9.3 mmol/l), low density lipoprotein cholesterol 236 +/- 18 mg/dl (6.1 mmol/l), triglycerides 333 +/- 28 mg/dl (3.8 mmol/l), and lipoprotein Lp(a) 46 +/- 11 mg/dl]. After 1 year of 40 mg of fluvastatin, significant reductions of total cholesterol by 31% to 242 +/- 26 mg/dl (6.3 mmol/l) and low-density lipoprotein cholesterol by 29% to 162 +/- 12 mg/dl (4.2 mmol/l) were observed. Furthermore, triglyceride values were also lowered significantly by 19% to 268 +/- 21 mg/dl (3.1 mmol/l). Lipoprotein Lp(a) and high-density lipoprotein-cholesterol remained unchanged by fluvastatin. These improvements in lipid profile were maintained during the entire follow-up period of 1 year. There were no adverse events, and the slight increase in serum creatinine observed during the study was considered to be due to the primary renal disease. In conclusion, long- term administration of fluvastatin in patients with nephrotic syndrome appears to be an effective and safe treatment of the hyperlipidemia associated with this disorder. PMID- 10460941 TI - Correlations among expression of glomerular intercellular adhesion molecule 1 (ICAM-1), levels of serum soluble ICAM-1, and renal histopathology in patients with IgA nephropathy. AB - The purpose of the present study was to evaluate the correlations among expression of intercellular adhesion molecule 1 (ICAM-1) in glomeruli, levels of soluble ICAM-1 (sICAM-1) in sera, and renal injuries in patients with IgA nephropathy. The levels of sICAM-1 in sera from 27 patients with IgA nephropathy and 7 healthy controls were measured by the human soluble ICAM-1 immunoassay. The expression of ICAM-1 in glomeruli was detected by indirect immunofluorescence. We observed marked expression of ICAM-1 in glomerular capillary walls and mesangial areas in patients with advanced-stage, but not in those with mild IgA nephropathy. Since the histopathological changes in the advanced stage of this disease were characterized by diffuse mesangial cell proliferation and tubulointerstitial injury, the expression of ICAM-1 in the glomeruli may be of value in evaluating the degree of renal lesions in patients with IgA nephropathy. However, there was no significant change in the levels of serum sICAM-1 among mild-stage and advanced-stage patients and healthy controls. It appears that the measurement of serum sICAM-1 is not useful in evaluating the degree of renal injuries in patients with IgA nephropathy. PMID- 10460942 TI - Protein intake seems to respond to increases in Kt/V despite baseline Kt/V greater than 1.2. AB - The nutritional status is a strong predictor of outcome in hemodialysis patients. Adequate delivery of dialysis is necessary for hemodialysis patients to maintain their protein nutrition. In general, a single-pool Kt/V of 1.2 has been considered adequate dialysis. We recently decided to maximize the blood flow during hemodialysis in all of our patients; this enabled us to increase the dose of delivered dialysis in those patients who were not initially utilizing their maximum blood flow. There were 18 patients who already received a Kt/V greater than 1.2. We were able to increase Kt/V even further in 10 of them, resulting in a significant increase in nPCR and a trend to increase serum albumin over the next 6 months of follow-up. The mean normalized protein catabolic rate (nPCR) and serum albumin remained unchanged in the remaining 8 patients whose Kt/V could not be increased. Our data provide evidence that protein intake in hemodialysis patients will increase with an increase in delivered dialysis above the level generally considered to be adequate. PMID- 10460943 TI - Effect of amlodipine therapy on the monoclonal antibody 3G8-induced calcium signal in polymorphonuclear leukocytes of hemodialysis patients. AB - Chronic renal failure is associated with impaired phagocytosis. This was attributed to the PTH-induced elevation of basal levels of [Ca2+]i of polymorphonuclear leukocytes (PMNLs) and to the small calcium transient induced by the ligation of the Fcgamma RIII receptors of these cells with 3G8 monoclonal antibodies. The blocking of the action of PTH on the PMNLs of patients with chronic renal failure by their treatment with a calcium channel blocker normalized the basal levels of [Ca2+]i of the PMNLs and reversed the defect in their phagocytic property. It is not known whether such therapy would also restore the calcium transient in the PMNLs in response to 3G8 monoclonal antibody to normal. We examined this issue in 12 normal subjects and 18 hemodialysis patients; 9 of them were treated with the calcium channel blocker, amlodipine, and the other 9 did not receive such therapy. The treatment with amlodipine normalized [Ca2+]i of PMNLs as well as the calcium transient in response to 3G8 monoclonal antibody and reversed the defect in their phagocytosis. It is concluded that chronic renal failure is associated with deranged calcium homeostasis of PMNLs which causes abnormalities in the function of Fcgamma RIII receptors and consequently results in impaired phagocytosis. Therapy with a calcium channel blocker can reverse all these derangements in metabolism and function of PMNLs. PMID- 10460944 TI - Serum hepatocyte growth factor levels in patients with renal diseases. AB - The serum levels of hepatocyte growth factor (HGF) were determined in patients with various renal diseases. In patients with acute-phase acute renal failure (ARF) and chronic tubulointerstitial nephritis (chronic TIN), the serum HGF levels were 0.55 +/- 0.24 and 0.44 +/- 0.37 ng/ml (mean +/- SD), respectively, and were significantly higher than that in the control group (0.12 +/- 0.12 ng/ml). The serum HGF level tended to be high also in patients with active-phase steroid-sensitive nephrotic syndrome (SSNS). The serum levels of HGF were not elevated in patients with IgA nephropathy (IgAN), Henoch-Schonlein purpura nephritis (HSPN), membranoproliferative glomerulonephritis (MPGN), poststreptococcal acute glomerulonephritis (PSAGN), unilateral renal atrophy, unilateral nephrectomy, or proximal tubular dysfunction. These observations suggest that glomerular disorders cause no apparent elevation of the serum HGF level, and that elevation of the serum HGF level may be associated with tubulointerstitial damage in renal diseases. PMID- 10460945 TI - Membranoproliferative glomerulonephritis with subendothelial deposits (type 1) associated with hepatitis G virus infection in a renal transplant recipient. AB - BACKGROUND: Infection with hepatitis B virus (HBV) or hepatitis C virus (HCV) is a well-known etiology for membranoproliferative glomerulonephritis (MPGN) with subendothelial deposits (MPGN type 1). MATERIAL AND METHODS: The newly discovered hepatitis G virus (HGV) is currently under active investigation. We report the first case of de novo MPGN type 1 associated with HGV infection in a young male renal transplant recipient who manifested glomerulonephritis (GN) with proteinuria 7 years after transplant, and whose original disease was chronic obstructive pyelonephritis secondary to nephrolithiasis. RESULTS: Serum markers for HBV and HCV infections were negative. HGV infection was detected by specific double-nested reverse transcriptase-polymerase chain reaction (RT-PCR) in sera (positive HGV viremia) 2.5 years after renal transplantation. By specific in situ RT-PCR, the presence of the HGV genome was detected in peripheral blood mononuclear cells and in the kidney biopsy (glomeruli and tubules), but not in the liver. CONCLUSION: This report adds new information on the role of HGV infection in the occurrence of de novo GN (MPGN type 1) in renal transplantation. PMID- 10460946 TI - Dialysis-associated renal cystic disease resembling autosomal dominant polycystic kidney disease: a report of two cases. AB - Acquired renal cystic disease is common in patients receiving dialysis. Characteristically, the kidneys are small or, less often, normal in size, and the cysts are usually less than 0.6 cm in diameter. We present here 2 patients who, after 5 and 7 years on hemodialysis, developed marked renal enlargement, with large cysts in the kidneys and, in 1 patient, in the liver as well; the appearance on ultrasonography and computed tomography was indistinguishable from autosomal dominant polycystic kidney disease. Before starting dialysis the first patient was a 19-year-old man who developed renal shutdown from crescentic glomerulonephritis, and the second patient was a 33-year-old man who developed end-stage renal failure from malignant hypertension. Neither patient had renal cysts at the onset of end-stage renal failure. PMID- 10460947 TI - A rapidly progressive cataract in a patient with autoimmune hypoparathyroidism and acute liver and renal failure. AB - Cataract is a well-known complication of hypoparathyroidism, albeit the mechanism is obscure. The progression of cataract is typically slow in patients with idiopathic hypoparathyroidism. We describe a case of an extremely rapid evolution of typical hypocalcemic cataracts in a patient with familial autoimmune hypoparathyroidism during acute idiopathic hepatic and renal failure, while serum calcium and phosphorus were unbalanced. Physicians and ophthalmologists must be aware of cataracts developing rapidly in the setting of such metabolic derangements. PMID- 10460948 TI - Chronic rheumatoid arthritis complicated by myeloperoxidase antineutrophil cytoplasmic antibody-associated nephritis. AB - We report a myeloperoxidase antineutrophil cytoplasmic antibody-positive rheumatoid arthritis patient who developed necrotizing crescentic glomerulonephritis. Steroid therapy was given combined with an immunosuppressant agent, and double-filtration plasmapheresis was started with the aim of removing antibodies from the blood. This therapeutic regimen was found to be useful. PMID- 10460949 TI - Mesangial cell proliferation in long-term streptozotocin-induced diabetes mellitus in the rat and the renoprotective activity of heparin. AB - At present, it is not clear whether mesangial proliferation underlies mesangial expansion in diabetic nephropathy. To address this issue and the relationship between heparin's renoprotective and antimitogenic activities, we studied three streptozotocin-induced diabetic rat groups 5 and 12 months after diabetes induction: two groups were administered a modified heparin, each with a different protocol, and two healthy rat groups, one of which was treated with the same heparin, served as controls. Untreated diabetic animals developed clear evidence of nephropathy, namely expansion of the glomerular extracellular matrix, as expressed by glomerular basement membrane thickening, and increased mesangial deposition of type IV collagen. These alterations were prevented/cured by heparin treatment. Kidney sections were processed immunohistochemically for proliferating cell nuclear antigen and smooth muscle alpha-actin which is expressed only by proliferating mesangial cells. The number of proliferating cell nuclear antigen positive nuclei and alpha-actin-positive cells per glomerulus did not differ between groups at both 5 and 12 months. In conclusion, there is no evidence that mesangial proliferation is increased in late experimental diabetic nephropathy, and heparin seems to be renoprotective through mechanisms other than antiproliferation. PMID- 10460950 TI - Influence of nasopharyngeal CPAP on breathing pattern and incidence of apnoeas in preterm infants. AB - Continuous positive airway pressure (CPAP) applied by endotracheal (ET) tube in the pharynx (nasopharyngeal CPAP, n-CPAP) is widely used for the treatment of mild respiratory distress syndrome and of apnoeas of prematurity. Effects on breathing pattern and on different types of apnoeas are not fully understood. We wanted to know the effect of discontinuing n-CPAP on the respiratory rate, apnoeas and bradycardia/desaturation events. Thirteen prematurely born infants with bradycardia and/or desaturation events were studied when weaning from n-CPAP was clinically considered. Polygraphic studies were performed for 2 h during n CPAP therapy and for 2 h without CPAP. Nasal flow was measured by registering expiratory pCO(2) at the free nostril opening. During n-CPAP, the respiratory rate was significantly lower, there were fewer obstructive apnoeas, more short central apnoeas (6-9 s) and less severe apnoea-associated desaturations. During n CPAP, the infants spent significantly more time in a state of quiet breathing. PMID- 10460951 TI - Functional and antigenic concentrations of alpha-1-proteinase inhibitor after administration for the prevention of chronic lung disease of prematurity. AB - OBJECTIVE AND METHODS: Alpha-1-proteinase inhibitor (A1PI) supplementation has been used in adults with inherited alpha-1-antitrypsin (A1AT) deficiency to impede the development of emphysema. A1PI supplementation may also be useful for protecting premature neonates who receive mechanical ventilation from the development of chronic lung disease (CLD). However, the pharmacokinetics of exogenous A1PI in this population are unknown. We attempted to determine the disposition of A1PI in premature infants with birth weight 600-1,250 g who received 60 mg/kg on days 0, 4, 7 and 14 in a randomized, placebo-controlled, double-blind trial. Functional and antigenic plasma concentrations of A1PI were measured at specified time points. RESULTS: On both functional and antigenic assays, concentrations began in the normal adult range and rose from day 0 to 10 then fell slightly, but remained above initial values. The concentrations were not significantly different between the treatment and placebo groups. CONCLUSIONS: The results of this study indicate that neonatal pharmacokinetics of A1PI differ markedly from those of the adult. Total plasma clearance of exogenous A1PI seems high in the ventilated premature neonate. Higher or more frequent doses may be necessary to maintain A1PI plasma concentrations above baseline. PMID- 10460952 TI - Activities of some glycosaminoglycan- degrading enzymes in Wharton's jelly and their alteration in EPH-gestosis (Pre-eclampsia). AB - Oedema, proteinuria, hypertension (EPH)-gestosis (pre-eclampsia) is associated with a premature replacement of hyaluronic acid by sulphated glycosaminoglycans (GAGs), both in the umbilical cord arteries and in Wharton's jelly. It may be concluded from our previous report that such a phenomenon may be the result of reduction in degradation of these compounds. In order to support such a conclusion the activities of GAG-degrading enzymes in normal umbilical cord arteries and those taken from newborns delivered by mothers with EPH-gestosis were compared. It was found that EPH-gestosis results in a significant reduction in the activities of neutral endoglycosidases degrading most of the sulphated GAGs (except keratan sulphate). In the case of acidic endoglycosidases, no characteristic alterations have been found. Only the activity of heparan sulphate degrading endoglycosidase significantly decreased. In contrast to the above mentioned endoglycosidases, the activities of arylsulphatase B and 6-sulphatase distinctly increased. The decrease in the activities of endoglycosidases are thought to be responsible for EPH-gestosis-associated accumulation of sulphated GAGs in extracellular matrix of Wharton's jelly. This leads to the suspicion that EPH-gestosis-induced changes in the GAGs composition may alter the fibrillogenesis conditions in Wharton's jelly. The sulphated GAGs accumulated in Wharton's jelly may interact with some growth factors which modify the myofibroblasts' proliferation, gene expression, protein biosynthesis and other processes. A significance of EPH-gestosis-induced alteration in Wharton's jelly is discussed. PMID- 10460953 TI - Effects of hypoxia and reoxygenation with 21% and 100%-oxygen on cerebral nitric oxide concentration and microcirculation in newborn piglets. AB - Bioelectric sensors for continuous registration of nitric oxide (NO) concentrations in tissues provide a new tool for invasive measurement of this gaseous molecule. This study sought to validate cerebral NO measurements using an amperiometric sensor. A series of experiments in 1- to 3-day-old piglets was carried out to study the response of NO and microcirculation during hypoxia (F(i)O(2) 0.06) and reoxygenation with 100% and 21% oxygen. Two-channel laser Doppler flowmetry was performed in the forebrain cortex. Significant decreases of NO levels were observed immediately after induction of hypoxemia (p < 0.05). During reoxygenation with 21 or 100% O(2) for 30 min, NO increased significantly compared to the values at the end of hypoxia (p < 0.05). The increase of NO levels in the 100% oxygen group was greater than the increase in the 21% oxygen group (p < 0. 05). There were no significant differences between the two groups during the following 3.5 h of observation. A significant increase in CBF was found in the first 2 min of hypoxia (p < 0.05), it then continued to fall to values significantly lower than baseline values at the end of hypoxemia (p < 0.05). During reoxygenation CBF normalised and there were consistent but no significant differences between the two reoxygenation groups. We conclude that NO concentration decreased during the course of hypoxia. Hypoxia-induced cerebral hyperaemia occurred in spite of significantly lower NO concentrations. Reoxygenation with 21 or 100% O(2) restored CBF in both groups similarly, although values were higher after reoxygenation with 100% O(2) compared to air. In fact, reoxygenation with 100% O(2) led to supranormal levels of NO by contrast to 21% O(2). PMID- 10460954 TI - Effect of increased brain GABA concentrations on breathing in unanesthetized newborn rabbits. AB - Evidence suggests that gamma-aminobutyric acid (GABA) is involved in control of breathing and in the hypoxia-related ventilatory depression in newborns. However, this evidence is obtained mainly from studies on anesthetized animals. Because anesthesia may interfere with the GABA system, the objectives of our study were to examine effects of GABA on ventilation (V(E)) and ventilatory response to hypoxia and to reveal effects of repeated hypoxia on GABA concentrations in unanesthetized newborns. The study was performed in rabbits in two age groups: 1 3 days old (group I) and 10-14 days old (group II). To increase brain endogenous GABA concentrations, rabbits were injected with an inhibitor of GABA transaminase, aminooxyacetic acid (AOAA; 20 mg/kg i.p.). To prevent postmortem formation of GABA, at the end of experiments the rabbits received an inhibitor of glutamic acid decarboxylase, IP-3-mercaptopropionic acid (100 mg/kg i.p.). Animals were studied in normoxia alone, or they were exposed for 15 min to 8% O(2) before and 10 and 35 min after saline or AOAA. GABA concentrations were measured in brainstem, cerebrum, and cerebellum by means of a capillary electrophoresis. In group I, AOAA had no respiratory effects. In group II, AOAA decreased V(E), tidal volume, and mean inspiratory flow in normoxia and reversed V(E) decline during hypoxia 10 min after the injection, GABA concentrations were not age dependent and the highest in the brainstem. Repeated hypoxia increased the cerebellar GABA concentrations and had no effect in group I. These results imply that in unanesthetized rabbits, GABAergic neurotransmission in the respiratory control network becomes functional by the 2nd week of life, but it does not contribute to the biphasic ventilatory response to moderate hypoxia. In contrast, GABA-evoked block of the cerebellar inhibitory input during hypoxia may be responsible for the reversal of the V(E) decline in unanesthetized newborns. PMID- 10460955 TI - Regulation of myocardial beta-adrenergic receptor function in adult and neonatal rabbits. AB - beta-Adrenergic receptor (betaAR) desensitization is the decrease in response following sustained agonist stimulation by catecholamines. While developmental changes in betaAR response have been well documented in the mammalian heart, much less is known regarding the regulation of betaAR function in immature hearts. The purpose of the present study was to determine whether there are developmental differences in myocardial betaAR desensitization. We used an isolated heart preparation to examine the betaAR-mediated inotropic response before and after sustained exposure to 1 microM isoproterenol in adult and neonatal rabbits. We also assayed the adenylyl cyclase activity and performed radioligand-binding studies to determine betaAR characteristics in adult and neonatal ventricular tissues with and without exposure to isoproterenol. Both adult and neonatal rabbit hearts showed a concentration-dependent increase in systolic function, namely, isovolumic left ventricular developed pressure (LVDP) and maximal positive dP/dt of LVDP (dP/dt(max)) in response to isoproterenol. Adults, however, showed a significantly greater response than neonates. After sustained exposure to isoproterenol, the subsequent betaAR-mediated responses in LVDP and dP/dt(max) were significantly attenuated in adults, but much less so in neonates. The adenylyl cyclase activity in response to isoproterenol was significantly different between adult, but comparable in neonatal tissues both exposed or not to isoproterenol. The total betaAR density was higher in neonatal than in adult tissues without isoproterenol exposure, but there was no significant change in betaAR density in either group following isoproterenol exposure. In addition, isoproterenol exposure increased the amount of the inhibitory G protein in adult, but not neonatal tissues. Our results suggest that there were developmental differences in myocardial betaAR functional responses in betaAR desensitization. PMID- 10460957 TI - A comparison of clinical and radiographic caries diagnoses in posterior teeth of 12-year-old Lithuanian children. AB - Bite-wing radiography has been recommended for use in adolescents as clinical examinations alone may lead to underestimation of carious lesions in approximal and occlusal surfaces. The aim of this study was to describe the relationship between clinical and radiographic caries diagnoses among 12-year-old Lithuanians using a new clinical scoring system which differentiates between cavitated and non-cavitated caries lesions. Eight hundred and seventy-two children were examined clinically and two standardized posterior bite-wing radiographs were taken of each participant. Bite-wing radiography contributed significantly to the total number of lesions diagnosed only at the dentin level in approximal surfaces. Clinical examination performed better than radiographic examination at the non-cavitated/enamel level, particularly on occlusal surfaces. Less than 2% of the clinically sound surfaces were diagnosed with dentin lesions/fillings radiographically. When using the cavitated level of clinical diagnosis, the frequency of 'hidden' caries lesions increased from 1.9 to 2.9% in approximal surfaces, and from 1.7 to 5. 2% in occlusal surfaces. The intra-examiner reliability data for the clinical and radiographic recordings supported the conclusion of an additional diagnostic value of bite-wings only for approximal surfaces. The findings demonstrate that the diagnostic yield of bite-wing radiography is higher for approximal than for occlusal surfaces. The efficacy of bite-wings depends on the refinement of the clinical caries diagnostic criteria. 'Hidden' caries does not seem to be a major problem when the clinical caries diagnostic criteria include non-cavitated diagnoses. PMID- 10460956 TI - Caries prevention in a community-dwelling older population. AB - A clinical trial was conducted to compare the effect of different caries preventive strategies on caries progression in lower-income, ethnically diverse persons 60 years of age and older. Two hundred and ninety-seven subjects were randomized into one of five experimental groups. Group 1 received usual care from a public health department or a private practitioner. Group 2 received an educational program of 2 h duration implemented twice a year. Group 3 received the educational program plus a 0.12% chlorhexidine rinse weekly. Group 4 received the education and chlorhexidine interventions and a fluoride varnish application twice a year. Group 5 received all the above interventions as well as scaling and root planing every 6 months throughout the 3-year study. A carious event was defined as the onset of a carious lesion, a filling, or an extraction on a surface which was sound at baseline. Two hundred and one subjects remained in the study for the 3-year period. Groups that received usual intraoral procedures (groups 3, 4, and 5) had a 27% reduction for coronal caries events (p = 0.09) and 23% for root caries events (p = 0.15), when compared to the groups that received no intraoral procedures (groups 1 and 2). Routine preventive treatments may have had only a small-to-moderate effect upon caries development. PMID- 10460958 TI - The influence of specific foods and oral hygiene on the microflora of fissures and smooth surfaces of molar teeth: A 5-day study. AB - A group of 20 students, harbouring >10(4) mutans streptococci per millilitre of saliva, was enrolled into the study. Models for sampling, reproducibly, the dental plaque present in specific sites (fissure and smooth surface) on the dentition were developed and validated. Withdrawal of normal oral hygiene procedures for only 1 day resulted in approximately 10-fold increases in the number of micro-organisms recovered from both sites. The effect of supplementing the subjects' diets with particular food items given 5 times per day [lemonade (5.8% w/v sugars, 250 ml), biscuits (digestive biscuits, 67.6% w/w carbohydrate of which 22% w/w was sugars and 45.6% w/w was starch), caramel toffees and sugar lumps] on the number of micro-organisms recovered and on the composition of the flora at both sites was determined. Dental plaque samples were taken after 5 days and it was found that supplementation of the diet with toffee and sugar lumps resulted in significantly more micro-organisms at both sampling sites. The supplementation of the diets with lemonade or biscuits did not significantly alter the numbers of micro-organisms recovered from either site. The percentage composition of the plaque samples from both dental sites remained relatively unaffected by oral hygiene although there were lower levels of mutans streptococci which might be related to the use of an antimicrobial toothpaste containing fluoride, triclosan and zinc citrate. This study suggests that the cariogenicity of certain sucrose-containing foods may, in part, be due to the enhancement of plaque accumulation in addition to other effects on the percentage composition of the plaque which may become manifest on prolonged usage of these dietary foodstuffs. These observations are consistent with dietary survey findings which often find consumption of confectionery related to caries experience or incidence. PMID- 10460959 TI - Distribution of three cariogenic bacteria in secondary carious lesions around amalgam restorations. AB - Secondary dental caries remains an unresolved problem in dentistry and little is known of its microbial etiology. The purpose of this study was to compare the distribution of the three most suspected cariogenic groups of bacteria, mutans streptococci. Actinomyces naeslundii genospecies 2 and lactobacilli, in natural secondary caries around amalgam restorations. Extracted teeth with secondary caries were sectioned to obtain three samples that were randomly distributed to three different groups. Each group was immunolabeled with antibodies to either Streptococcus mutans, A. naeslundii genospecies 2 or Lactobacillus casei and subsequently labeled with secondary fluorescent antibodies. All samples were analyzed three-dimensionally using confocal microscopy. The results indicated that the three different bacteria were widely present and could have an important role in the development of secondary caries around amalgam restorations. PMID- 10460960 TI - Inhibitory effect of antibacterial resin composite against Streptococcus mutans. AB - Dental resin composites with antibacterial activity may be useful for preventing the secondary caries frequently seen around restorations. Three types of silver supported antibacterial materials (Novaron, Amenitop and AIS) inhibited the growth of the major oral pathogen Streptococcus mutans. Minimum inhibitory amounts in suspensions of Novaron, Amenitop and AIS against S. mutans were 40, 30 and 400 microg/ml, respectively. These antibacterial materials were incorporated into TEGDMA-UDMA-based light-activated resin composite, and the antibacterial activities of these composites were examined. Composites incorporating 5 wt% (N 5) or more of Novaron and 7 wt% (AM-7) or more of Amenitop inhibited the growth of S. mutans, whereas composites incorporating up to 10 wt% of AIS did not. No significant difference in either compressive or flexural strength was observed between the control and N-5 composites after 1 day and 6 months of storage in water. However, for AM-5 composite, there was a significant difference in either strength parameter between the two immersion periods. There was no or extremely little release of silver ions from the N-5 and AM-5 composites after 1 day or 6 months of immersion in water. These results indicated that a light-activated resin composite incorporating silver-supported antibacterial material such as Novaron may be clinically useful due to its inhibitory effect against S. mutans and favorable mechanical properties. PMID- 10460961 TI - Influence of in vivo formed salivary pellicle on enamel erosion. AB - This study assessed the protective effect of the salivary pellicle formed in vivo during 24 h or 7 days against demineralization of bovine enamel caused by citric acid. In addition, the influence of acid treatment on the behavior of the pellicle was investigated. Enamel specimens with and without in vivo pellicles were immersed in citric acid (0.1, 1.0%) over 30, 60, and 300 s, and processed for scanning (SEM) and transmission electron microscopy (TEM), as well as for measurement of surface microhardness (SMH). Specimens coated with the in vivo formed pellicles revealed less extensive erosive demineralization of the enamel surface compared to uncovered enamel specimens. SEM analysis and SMH results did not indicate distinct differences between erosive surface alterations on enamel slabs covered with 24-hour pellicles and on those covered with 7-day pellicles. TEM analysis showed that the pellicle layer was dissolved in part from the enamel surface due to acid exposure. However, pellicle residues could be detected by TEM in all specimens, even after 5-min exposure to 1.0% citric acid. It is concluded that the in vivo salivary pellicle can resist the acidic action to some extent and provides protection to the underlying enamel surface against erosive destruction caused by short-term action of citric acid. PMID- 10460962 TI - A two-film versus a four-film bite-wing examination for caries diagnosis in adults. AB - The aim of this study was to compare the caries diagnostic outcome of a two-film and a four-film routine bite-wing examination undertaken in an adult population. A digital bite-wing examination using two and four films, respectively, was performed in 174 patients. The radiographs from the two examinations were assessed jointly by 2 observers for the number of imaged surfaces in the area from the distal surface of the canine to the distal surface of the third molar and for the detection of carious lesions. Significantly more surfaces were available for examination by the four-film than by the two-film examination. Overlapping surfaces were recorded in 14 and 10%, respectively, of the surfaces imaged by the two-film and four-film set. The prevalence of primary enamel and dentinal caries was 6.8% with the two-film and 5.6% with the four-film method. In 1, 684 surfaces only imaged by the four-film method caries was detected in 20 surfaces. If these extra lesions were added to the number of lesions found by the two-film method the total disease prevalence using the last-mentioned method would increase by 2 per thousand. The majority (92-99%) of all recorded lesions were detected in surfaces located between the occlusal surface of the first premolar and the mesial surface of the third molar. The little additional diagnostic outcome obtained by the use of four instead of two films did not seem to match the extra resources (double patient charge and radiation dose, and extended scan/developing and recording time for the dentist) connected to the four-film method. Thus, it may be recommended that a routine bite-wing examination undertaken in young adults should be performed with only two films: one in each side of the mouth, placed behind the premolars and the first and second molars. PMID- 10460963 TI - Dyract versus Tytin Class II restorations in primary molars: 36 months evaluation. AB - Due to the changed treatment approach of proximal caries and the amalgam controversy, clinicians are in search for new materials. The aim of the present study was to compare amalgam with an adhesive material in deciduous molars in a clinical, split-mouth design study. At baseline 30 polyacid modified composite (Dyract) and 30 amalgam (Tytin) restorations were placed in primary molars, of which 24 and 17 could be evaluated after 24 and 36 months, respectively. Modified USPHS criteria were used for clinical evaluation every 6 months. Annual bite-wing radiographs were taken for evaluation of recurrent caries and cervical gap formation. In the present study, for Dyract as well as for Tytin restorations, low rates of recurrent caries were found, while Dyract restorations showed a better marginal adaptation and surface texture compared to Tytin restorations. In the Dyract group more radiolucencies were found at baseline. In both groups no patient complaint or pain was reported related to the radiolucencies. After 3 years the colour of Dyract was not comparable to the original. For Dyract no excessive wear was noticed compared to enamel. During the study one Dyract (recurrent caries: 18 months) and two Tytin (pulpal aetiology: 6 months, recurrent caries: 36 months) restorations had to be replaced. Even though the restorations were placed in caries risk children, at the 36 months' evaluation of this clinical study, the results indicate that Dyract can be an alternative for Tytin in the primary dentition. PMID- 10460964 TI - Effect of Apis mellifera propolis from two Brazilian regions on caries development in desalivated rats. AB - The purpose of the present study was to evaluate the effect of Apis mellifera propolis collected from two regions of Brazil on caries development in desalivated rats. Ethanolic extracts of propolis (EEP) were prepared from crude propolis samples collected in Minas Gerais state (MG), southeastern Brazil, and Rio Grande do Sul state (RS), southern Brazil. The flavonoid composition of EEP was analyzed by high-performance thin-layer chromatography (HPTLC) and reversed phase high-performance liquid chromatography (HPLC). For the animal study, 30 specific pathogen-free Wistar rats were infected with Streptococcus sobrinus 6715 and surgically desalivated. The rats were randomly divided into three groups which were treated with 80% ethanol (control), EEP from MG and EEP from RS. The animals were placed in a Konig-Hofer programmed feeder and received 17 meals of diet 2000 daily at hourly intervals. The solutions were applied on the rat molars (25 microl on molars of each quadrant) twice a day, by using graduate syringes. After 3 weeks, the animals were killed by CO(2) asphyxiation. For microbial assessment, the left jaw was removed and sonicated in 154 mM NaCl solution. Dental caries was evaluated according to Larson's modification of Keyes' system. The HPTLC patterns and HPLC profiles demonstrated that both quality and quantity of flavonoid aglycones of EEP from MG were different compared to EEP from RS. In general, it is apparent that EEP from RS contained the highest concentrations of pinocembrin, chrysin, acacetin and galangin. The group of animals treated with EEP from RS showed the lowest smooth-surface and sulcal caries scores as well as less caries severity in smooth-surface and sulcal lesions, and these data were statistically different when compared with the control group. The group treated with EEP from MG only demonstrated a significant difference in the severity of sulcal lesions when compared to the control group. The percentage of S. sobrinus was lower in the groups treated with EEP, but did not differ statistically from the control group. The results showed that the cariostatic effect of propolis depends on its composition, and consequently the region of collection of propolis samples. PMID- 10460965 TI - Toothbrushing habits and caries experience. PMID- 10460966 TI - In vitro adhesion to enamel and in vivo colonization of tooth surfaces by Lactobacilli from a bio-yoghurt. PMID- 10460967 TI - Involvement of sulfated proteoglycans in embryonic brain expansion at earliest stages of development in rat embryos. AB - The expansive force generated by the positive pressure of the neural tube fluid confined inside the brain vesicles has been shown to be a key factor during the earliest stages of brain morphogenesis and development of chick embryos. In a previous study, we demonstrated the existence of an intracavity extracellular matrix rich in condroitin sulfate in this species, which could be involved in the regulation of the expansive process. In this report, scanning electron microscopy and immunohistochemistry show that, after neurulation, a similar extracellular matrix rich in chondroitin sulfate is present inside the brain vesicles of rat embryos during early enlargement of the brain anlage. In vitro treatment of rat embryos with beta-D-xyloside (a chemical compound which disrupts chondroitin sulfate synthesis) shows that changes in intralumen chondroitin sulfate concentration are accompanied by significant changes in brain anlage growth. These results support the hypothesis that intracerebral chondroitin sulfate plays a relevant role in the regulation of the expansive process of the brain anlage of rat embryos, and could represent a general mechanism in the early brain development of avian and mammalian embryos. PMID- 10460968 TI - Transcript-specific mRNA trafficking based on the distribution of coexpressed myosin isoforms. AB - mRNAs encoding four myosin heavy chain (MHC) isoforms were localized in rat skeletal muscle fibers by in situ hybridization. The ratio of MHC transcript signal in the fiber core compared to the fiber periphery was quantified using image analysis. Two distinct patterns of subcellular localization were observed. Type 1 (beta-cardiac) and type 2A MHC mRNAs were located preferentially in the muscle fiber periphery, while type 2B and type 2X mRNAs were distributed homogeneously across the fiber cross section. Since most normal muscle fibers express only a single MHC isoform, this difference in mRNA distribution could reflect either variation in the localization of the synthetic apparatus across different fiber types or differences in the trafficking of different MHC transcripts. To examine the basis for the observed differential distribution in normal muscles, mRNA distribution was assessed in muscle fibers that coexpressed multiple isoforms of the fast MHCs (i.e. types 2A, 2X and 2B), which occurred either in the combination type 2A/2X or type 2X/2B. The quantitative mRNA distribution seen in muscle fibers expressing a single isoform was not significantly different compared to that observed for mRNAs coexpressed in the same fiber (p > 0.6). Given the size similarity and homology of our riboprobes, these data suggest that their subcellular localization may be determined by relatively small differences in the sequences of the mRNAs, perhaps by differential binding of RNA sequence motifs to cytoskeletal elements. PMID- 10460969 TI - Connexin43 expression in network-forming cells at the submucosal-muscular border of guinea pig and dog colon. AB - The tissue-specific expression of connexin subtypes in gap junctions between the interstitial cells and smooth muscle cells in the submuscular plexus of the colon has a functional importance in relation to intestinal pacemaker activity. Immunocytochemical observations of two types of connexin molecules, connexin43 and connexin45, were made with a confocal laser scanning microscope on cryosections of freshly frozen dog, guinea pig, mouse and rat proximal colon. Connexin43 immunoreactivity appeared as a series of dots along the submuscular plexus of guinea pig and dog. In contrast, connexin43 immunoreactivity was not found in mouse and rat colon. Connexin43 immunoreactivity was not observed in the colon muscular layer in the four animal species examined. In double-stained materials with a marker for either vimentin or smooth muscle actin, connexin43 immunoreactivity was colocalized with vimentin immunoreactivity, whereas it was not with either smooth muscle actin immunoreactivity or phalloidin reactivity. This indicated that the connexin43-expressing cells possess a vimentin-positive fibroblast-like nature rather than a smooth muscle-like one. In addition, in guinea pig colon, connexin43 immunoreactivity colocalized with c-Kit immunoreactivity. In conclusion, network-forming cells are connected by connexin43 gap junctions in the submuscular plexus of guinea pig and dog colon, most likely indicating that interstitial cells act as an intestinal pacemaker and conductive system. PMID- 10460970 TI - Structural and ultrastructural study of GH, PRL and SMT cells in male goat by immunocytochemical methods. AB - The structural and ultrastructural characteristics of adenohypophyseal growth hormone (GH)-, prolactin (PRL)- and GH-PRL (SMT)-secreting cells were studied using immunocytochemical techniques in two normal and one lactating male goat. SMT cells were found in both types of males, which showed the same characteristics as those reported for the female. PRL and SMT cells were more frequent in the lactating male, which reflects their greater galactopoietic activity. GH cells did not seem to influence this process significantly. PMID- 10460971 TI - Rat model of Achilles tendon disorder. A pilot study. AB - Three-month-old male rats were subjected 3 times weekly for 1 h to eccentric exercise of one triceps surae muscle (30 stimulations/min) under general anesthesia in order to induce Achilles tendon disorder corresponding to paratenonitis and tendinosis in man. Net muscle work during the sessions ranged between 0.67 and 4.37 mJ (mean 1.72, SD 0.77). After 9 and 13 sessions, respectively, 2 rats started to show gait alterations during the functional test which was performed 2-3 times weekly. These rats were killed after additional sessions which showed a worsening of the limp. The other trained rats and controls did not limp and were killed after 7-11 weeks. Histologic evaluation of the Achilles tendons from the exercised limb showed in the majority of the cases hypervascularization, increased number of nerve filaments and increased immunoreactivity for substance P and calcitonin gene-related peptide. The tendons from the nonstimulated limb looked normal. The distribution of collagen types I and II appeared normal in the tendon and its insertion to the calcaneus. Inflammation of the epi- and paratenon could be provoked in the rat, but tendon changes corresponding to chronic tendinosis did not develop within 11 weeks with the used training regime. The clinical relevance of this model for chronic tendon disease needs to be evaluated further. PMID- 10460972 TI - Innervation of human trigeminal nerve blood vessels. AB - Film preparations and histological sections of human trigeminal nerve impregnated with silver nitrate and treated according to Falck-Hillarp revealed a rich innervation within the intraneural blood vessels. Highly diverse and complex neural interconnections were noted in the arterioles and venules of the trigeminal ganglion capsule as well as in the epineurium and external layers of the perineurium of the trigeminal nerve branches. Neural plexuses were detected on the vessel walls within these layers. Sensory innervation of the neural blood vessels was mainly relegated to polyvalent tissue-vascular receptors with adrenergic neural plexuses encountered on the walls of the intraneural vessels. PMID- 10460973 TI - Sympathetic and sensory innervations are heterogeneously distributed in relation to the blood vessels at the extensor carpi radialis brevis muscle origin of man. AB - By using immunohistochemistry and antibodies to a general nerve marker, protein gene product (PGP) 9.5, the overall innervation at the extensor carpi radialis brevis (ECRB) muscle origin was investigated in patients with tennis elbow and in healthy controls. The autonomic innervation was studied by using antibodies to neuropeptide Y (NPY), tyrosine hydroxylase (TH), and vasoactive intestinal peptide (VIP). The sensory innervation was visualized by using antibodies to substance P and calcitonin gene-related peptide. PGP 9.5 immunoreactions were detected in association with small blood vessels and arteries and within nerve bundles. There was, however, heterogeneity in the perivascular nerve fiber distribution since some blood vessels exhibited a high degree of PGP 9.5 innervation and some negligible or no such innervation at all. There was marked TH/NPY innervation in the walls of a subpopulation of the arteries, basically no VIP-containing nerves, and sensory innervation restricted to the small blood vessels. These observations show that the ECRB muscle origin is supplied with heterogeneously distributed sympathetic and sensory innervations and, furthermore, that there appears to be an imbalance between the vasoconstrictor and vasodilator innervations along the vascular tree in this region. PMID- 10460974 TI - National treatment systems in global perspective. AB - Drug policy development is mostly viewed as emerging within the nation state. Processes of diffusion of innovative policies have been neglected to a large extent. The comparative study of public policy has demonstrated, however, that diffusion is an important predictor of early policy adaptation. Thus, the analysis asks the general question of the relative importance of endogenous and exogenous effects on the development of drug policies in various countries. Specifically it describes the Swiss debate leading to the popular initiative on 'Youth Without Drugs' as well as the international reactions regarding its liberal outcome. Results of an expert survey show two broad types of reactions. There is one set of countries where chances for the introduction of limited heroin-prescription trials during the next 5 years are considered probable and a second set of countries which seems to be strictly status quo oriented. In the concluding section a model is suggested which systematically considers endogenous as well as exogenous predictors of 'soft' or 'hard' drug policy adoption. Results of a first tentative test of the model are encouraging for future empirical research on diffusion processes of drug policies. PMID- 10460975 TI - Drug treatment systems and policy frameworks: a comparative social policy perspective. AB - The article offers a discussion of drug treatment systems from a social policy perspective. Comparative studies of social policy have utilised typologies to aid analysis. These are briefly characterised in this article. Current analysis of social policy points to changes in the shape of policy regimes. How are these changes impacting on drug treatment systems? Key principles around which policy forms are organised are the desire to effect social control, the desire to meet human need, and the need to minimise harm from risks present in the environment. In practice, these principles operate in tension and different outcomes are observable in different societies. The article argues that, in spite of these historical and cultural differences, some convergence is observable in systems as they adapt to global influences. There remains however an urgent need for more detailed empirical research on policy responses in different countries, utilising the case-study approach adopted by the ISDRUTS collaborators. PMID- 10460976 TI - Drug treatment in contemporary anthropology and sociology. AB - The relatively slim social science literature on drug treatment is reviewed. Attention is paid to the institutionalization of treatment at the meso (local community or clinic) level and the micro (client) level rather than to the more commonly examined macro (societal or national) level. The inter-penetration across these levels of ideologies and practices around drugs is revealed through discussion of the targeting of certain populations for treatment, methods of client control in treatment settings, and the client's view of treatment. In the literature, drug users are often presented as passive individuals, subject to various forms of restrictive social control in therapeutic settings. Their perspective on the processes or efficacy of treatment is rarely sought. The concept of drug treatment as necessarily beneficial to clients is questioned in this paper. PMID- 10460977 TI - Drug misusers and their treatment in the Czech Republic: changing problems and changing structures. AB - This paper reviews recent developments in treatment of drug-related disorders in the Czech Republic after the socio-political changes in 1989. In the first part, a profile of drug users is given. In the second part, changes in the inpatient and outpatient treatment facilities are discussed. PMID- 10460978 TI - Drug treatment in the Baltic countries. AB - Illegal drug use was already a problem in the Baltic countries in the Soviet era, when the Iron Curtain was still in place. At independence the Baltic countries inherited drug treatment system, operated mainly by psychiatrists and based on inpatient system. As the Soviet treatment model was abandoned and professionals in the Baltic countries turned to the West, the drug treatment field became open to the Western drug treatment experience, especially from European countries and USA. Unexpectedly, they were to be exposed to different and often conflicting concepts of treatment modalities, which usually competed with one another in claims of being the most effective and useful. The question remains open as to which programs will be developed to meet the growing demand for treatment in each country and how widely they will be adopted. PMID- 10460979 TI - Recent trends in drug treatment in Europe. AB - The article describes the recent developments in drug treatment systems in several European countries. The article is based on the up-dated papers delivered in the closing meeting of the ISDRUTS-project (International Study of the Drug Treatment Systems) in Lisbon, October 7-9, 1998. In the article latest trends in drug situation and drug-related harm in different countries are represented. Also recent changes in legal measures, the proceeding of the harm reduction measures, the situation with heroin trials, the implosion of the drug treatment system into the alcohol treatment system and alternatives to imprisonment and other diversion mechanisms for addicts are described. In the concluding chapter recent trends in drug treatment are analysed in the framework of the political climate in Europe. PMID- 10460980 TI - The contextualization of drug treatment - future research perspectives and concepts. AB - In the article the growing interest of studies on treatment systems rather than specific treatment modalities is taken as a starting point for a discussion and outlook on how the research agenda of drug treatment research will develop in the near future. The relative absence of research that provides substantial guidance in the search for treatment effects has opened up for studies on larger, 'natural' entities, which are examined from composite, and theoretically elaborated, standpoints. The article argues that there is much to be done when it comes to research on treatment from a client's perspective and that a development of that particular path is a necessity for a more profound description and understanding of treatment. The need for theoretical input is emphasized and exemplified with ideas from organizational theory and discussions related to the concept of globalization. Finally the article identifies a need for more attention to some definitional issues that must be addressed in order to empirically develop comparative research on drug treatment systems. PMID- 10460981 TI - [Changes of renal blood flow during organ-associated foot reflexology measured by color Doppler sonography]. AB - Using colour Doppler sonography blood flow changes of the right kidney during foot reflexology were determined in a placebo-controlled, double-blind, randomised study. 32 healthy young adults (17 women, 15 men) were randomly assigned to the verum or placebo group. The verum group received foot reflexology at zones corresponding to the right kidney, the placebo group was treated on other foot zones. Before, during and after foot reflexology the blood flow of three vessels of the right kidney was measured using colour Doppler sonography. Systolic peak velocity and end diastolic peak velocity were measured in cm/s, and the resistive index, a parameter of the vascular resistance, was calculated. The resistive index in the verum group showed a highly significant decrease (p /=5.5 micromol/l (upper decile of distribution of control values). Adjusted for maternal age, parity, and body mass-index, a second trimester elevation of homocyst(e)ine was associated with a 3. 2-fold increased risk of preeclampsia (adjusted OR = 3.2; 95% CI 1. 1-9.2; p = 0.030). There was evidence of a interaction between maternal adiposity (as indicated by her prepregnancy body mass index) and parity with second trimester elevations in serum homocyst(e)ine. Nulliparous women with elevated homocyst(e)ine levels experienced a 9.7-fold increased risk of preeclampsia as compared with multiparous women without homocyst(e)ine elevations (95% CI 2.1-14.1; p = 0.003). Women with a higher prepregnancy body mass index (>/=21.4 kg/m(2), or upper 50th percentile) and who also had elevated homocyst(e)ine levels, as compared with leaner women without homocyst(e)ine elevations were 6.9 times more likely to later develop preeclampsia (95% CI 1.4-32.1; p = 0.016). CONCLUSION: Our findings are consistent with other indications of vascular endothelial dysfunction predating clinical preeclampsia. Studies designed to examine the effect of dietary and/or pharmacological mediators of homocyst(e)ine metabolism in preeclampsia are warranted. PMID- 10461000 TI - Ploidy analysis and S-phase fraction determination by flow cytometry in anembryonic pregnancy and spontaneous abortions. AB - The nuclear DNA content of 20 anembryonic pregnancies was studied by flow cytometry from paraffin embedded tissue blocks. An abnormal amount of DNA content was found in 8 of the cases. This was a significantly higher percentage than encountered in spontaneous abortions studied by the same population (40 and 9%, respectively, p < 0.05). The S-phase fraction in anembryonic pregnancies was lower than in spontaneous abortions (22.4 +/- 12.7 and 35.4 +/- 6.8, respectively, p < 0.01). The results indicated that abnormal embryogenesis with grave chromosomal aberrations may play a major role in the etiology of anembryonic pregnancy. PMID- 10461001 TI - The chick embryo chorioallantoic membrane as a model to investigate the angiogenic properties of human endometrium. AB - The chick embryo chorioallantoic membrane (CAM) is an established in vivo angiogenesis assay. The aim of our study was to assess the angiogenic properties of endometrium and to quantitate the vascular response in an accurate way. Samples of proliferative endometrium (n = 17) and control mouse skin tissue (n = 8) were explanted onto the CAM at day 10 of incubation. Additional controls consisted of normal unmanipulated CAM (n = 12). Four days after grafting, photographs of the explant and the surrounding area were taken in ovo to measure the vascular density index (VDI). The VDI is a stereological estimate of vessel number and length, which was obtained by counting the intersections of vessels with a circular grid superimposed on a computerized image. Endometrium caused a significant increase in VDI as compared to both unmanipulated CAM (p < 0.001) and skin tissue as a control (p < 0.007). The intra-observer variability was 5.2%. This study demonstrates that the CAM assay is a suitable model to assess the angiogenic properties of endometrium. Furthermore, it allows detailed quantitation of the vascular response in an objective and reproducible way. Our findings suggest the CAM to be a promising model to study the role of angiogenesis in both normal human endometrium and diseases involving the endometrium. PMID- 10461002 TI - Comparative study of cervical laminar tents prior to extra-amniotic injection of ethacridine lactate (Rivanol) and a condom-nelathon catheter method for second trimester pregnancy interruption in Vietnam. AB - OBJECTIVE: To determine whether the regimen for termination of second-trimester pregnancies using laminaria tents 12-24 h prior to extra-amniotic ethacridine lactate (Rivanol) instillation, is more effective in shortening the insertion expulsion interval than the presently used method of abortion induction by a condom/Nelathon catheter. DESIGN: A prospective randomised comparative study was performed at Uong Bi General Hospital in Quang Ninh Province, Vietnam, on 91 women undergoing pregnancy termination in the second trimester. INTERVENTIONS: The subjects were randomly allocated to 2 treatment groups, receiving either the Nelathon catheter-condom method (n = 50) or by insertion of a laminaria tent into the cervical canal for preinduction, 13-29 h before extra-amniotic instillation of ethacridine lactate (n = 34). Seven subjects were not pretreated with the laminaria tent. MAIN OUTCOME MEASURES: The insertion-expulsion intervals and the incidence of side effects were assessed. RESULTS: The mean insertion-expulsion interval in the 2 groups was not significantly different (mean 27.5 +/- 16.1 and 26.4 +/- 16.4 h, respectively), calculating the insertion-expulsion interval from the start of active treatment, i.e. from the instillation of Rivanol or insertion of the Nelathon catheter and condom, until expulsion of the fetus. CONCLUSIONS: The laminaria-Rivanol method for pregnancy interruption is not more advantageous than the existing Nelathon catheter-condom method. Simple, successful and cost effective methods in achieving second-trimester abortion in the Vietnamese context have therefore to be identified and tested. PMID- 10461003 TI - Dyspareunia, back pain and chronic pelvic pain: the importance of this pain complex in gynecological practice and its relation with grandmultiparity and pelvic relaxation. AB - The aim of this study was to evaluate the correlation between severe dyspareunia, back pain, dysmenorrhea and chronic pelvic pain (CPP), and the relationship of each pain type with various sociodemographic factors, pelvic relaxation and high parity. Two hundred thirty-five premenstrual individuals were included. The prevalences of CPP, deep dyspareunia, dysmenorrhea and back pain were found to be 80.4, 30.6, 57.0 and 57.4%, respectively. Marriage duration was longer (p < 0. 01) and also parity was higher (p < 0.0001) in CPP cases than controls. However, none of the demographic factors had a significant correlation with dyspareunia, back pain and dysmenorrhea. CPP was correlated with both back pain (r = 0.18, p < 0.005) and dyspareunia (r = 0.19, p < 0.004). However, there was no correlation between back pain and dyspareunia. On the other hand, dysmenorrhea did not show a correlation with any pain types. While grandmultiparity had a significant effect on CPP (p < 0.0001), it did not have a significant effect on other pain types in a MANOVA model. CPP is very often seen in our population and it often makes a pain complex with dyspareunia and back pain. The prevention of grandmultiparity is important to decrease the incidence of CPP. PMID- 10461004 TI - Effect of ovarian involvement on the frequency of luteinized unruptured follicle in endometriosis. AB - OBJECTIVE: When we review the current literature on endometriosis and luteinized unruptured follicle (LUF), we see that most of the studies deal with only the association between LUF frequency and the severity of endometriosis. Our purpose was to evaluate the effect of ovarian involvement on LUF frequency in endometriosis and assess the relationship between endometriosis and LUF in infertile women. STUDY DESIGN: This study is a prospective analysis covering a total of 126 infertile women between 22 and 35 years of age who underwent diagnostic laparoscopy from September 1995 to August 1997 in the Department of Obstetrics and Gynecology at Suleyman Demirel University, Isparta, Turkey. Endometriosis was diagnosed in 58 of these patients. All had received a revised American Fertility Society staging score at the time of the laparoscopic diagnosis. Diagnosis of LUF was made when the following criteria were fulfilled: absence of ultrasonic signs of ovulation, and absence of an ovulation ostium on the follicle by laparoscopy, despite increased serum progesterone. Statistical evaluation was performed using chi2 test and Fisher's exact test where appropriate. RESULTS: The prevalence of LUF in mild, moderate and severe endometriosis cases was 13.3, 41.2 and 72.7%, respectively. The LUF frequencies were 45.9% in 37 endometriosis patients with ovarian involvement, 9.5% in 21 cases without ovarian involvement, and 5.9% in 68 cases without endometriosis. A statistically significant difference was observed between the LUF rate in the group with ovarian involvement and that without involvement (chi(2) = 8.06, p < 0.001). CONCLUSION: In summary, in this study we noted a significant increase in LUF frequency in endometriosis patients with ovarian involvement. PMID- 10461005 TI - Prolactin is an autocrine or paracrine growth factor for human myometrial and leiomyoma cells. AB - OBJECTIVE: To test the hypothesis that prolactin (PRL) acts as a mitogenic growth factor for human leiomyoma and myometrial cells. METHODS: To test this hypothesis, we performed three different types of experiments. First, we assessed whether exogenous PRL acted as a mitogen for cultured uterine smooth muscle cells. Second, we examined the role of endogenous PRL by assessing the cell number after exposure of the cultures to a neutralizing antibody to PRL. Finally, we examined both fresh tissues and cultured cells for expression of the PRL receptor messenger ribonucleic acid using the techniques of reverse-transcriptase polymerase chain reaction and Southern blotting. RESULTS: A significant suppression in cell number was seen after 5 days of culture for leiomyoma cells but not for myometrial cells after treatment with exogenous PRL. Both cell types showed a significant decrease in cell number after treatment with anti-PRL antibody. A 893-bp segment consistent with the cytoplasmic domain of the long form of the PRL receptor was amplified from both fresh and cultured tissues and confirmed by Southern blotting and sequencing. CONCLUSIONS: PRL appears to be an autocrine or paracrine growth factor for both leiomyoma and myometrial cells. However, there are some differences between tissues in their sensitivity to this growth factor. PMID- 10461006 TI - Clinical use of serum c-erbB-2 in patients with ovarian masses. AB - The c-erbB-2 (Her-2/neu) gene product has a large extracellular domain (ECD) and part of which could be identified in the serum. We measured the serum level of c erbB-2 ECD in 93 patients, who presented with ovarian masses, with an enzyme immunoassay test and an elevated level was found in 5.5, 16.7 and 38% of patients with benign, borderline and malignant ovarian neoplasms, respectively. This serum marker may reflect the overexpression of c-erbB-2 gene in tumor tissues, which is associated with poor prognosis. However, measurement of c-erbB-2 ECD when used alone or in combination with CA 125 is not useful in differentiating benign from malignant ovarian tumors. PMID- 10461007 TI - Supernumerary ovary found by ultrasonogram and FSH measurement after an extensive operation for a yolk sac tumor of the ovary. AB - A rare case of supernumerary ovary found by a transvaginal ultrasonogram and follicle-stimulating hormone (FSH) measurement is presented. The patient was a 32 year-old female who underwent an extensive operation for a yolk sac tumor of the ovary. An asymptomatic cystic tumor was found during follow-up. There was no evidence of recurrence or metastasis of the yolk sac tumor. Although histological confirmation was not possible because the patient refused removal of the mass, a diagnosis of supernumerary ovary was made because changes in the shape of the cystic mass completely correlated with changes in the serum FSH level. PMID- 10461008 TI - Endometrioid adenocarcinoma arising from adenomyosis. AB - In spite of many references to carcinoma arising from endometriosis at extrauterine sites, there are few documented cases of carcinoma developing in association with adenomyosis. We present 2 rare cases of adenocarcinoma arising from adenomyosis. The relationship between prior frequent estrogen use and carcinogenesis and the possible effects of chemotherapy and radiation therapy are reviewed. PMID- 10461009 TI - Re: Breast-feeding, pain and infection. PMID- 10461010 TI - Relative and absolute changes of activated platelets, microparticles and platelet aggregates after activation in vitro. AB - Standard flow cytometers provide relative numbers of activated platelets, microparticles, and platelet aggregates. With fluorescent beads it is now possible to determine absolute numbers. Whole blood and platelet-rich plasma were incubated with agonists (ADP, collagen, thrombin). CD62p expression, microparticle and platelet aggregate formation were measured. Flow-Count Fluorospheres((R)) were added to calculate absolute concentrations. After activation there was an increase in the percentage of CD62p-positive platelets. However, the total number of platelets decreased and therefore the absolute number of CD62p-positive platelets did not increase but decreased. The number of CD62p-positive platelets decreased not as much as the number of CD62p-negative platelets, which explains why the relative percentage of CD62p-positive platelets increased. A similar increase in percent and decrease in absolute counts was found for microparticles. Platelet aggregates increased both in relative and absolute numbers. These results suggest that the detection of activated platelets by flow cytometry has to be complemented by the determination of the absolute concentrations to avoid misinterpretation. PMID- 10461011 TI - The procoagulant effect of thrombin on fibrin(ogen)-bound platelets. AB - In a final stage of activation, platelets become procoagulant because of the appearance of phosphatidylserine (PS) at the membrane outer surface. This PS exposure requires a rise in cytosolic [Ca(2+)](i), is accompanied by formation of membrane blebs, and stimulates the formation of thrombin from its precursor prothrombin. Here, we investigated whether thrombin, as a potent platelet agonist, can induce this procoagulant response in plasma-free platelets interacting with fibrin or fibrinogen through their integrin alpha(IIb)beta(3) receptors. First, in platelets that were stimulated to spread over fibrin or fibrinogen surfaces with adrenaline, addition of thrombin and CaCl(2) caused a potent Ca(2+) signal that in about 30% of the cells was accompanied by exposure of PS. At low doses, integrin alpha(IIb)beta(3) receptor antagonist (RGD peptide) inhibited platelet spreading as well as thrombin-evoked PS exposure. Second, in platelet-fibrinogen microaggregates that were preformed in the presence of adrenaline, thrombin/CaCl(2) induced PS exposure and bleb formation of about 35% of the cells. Third, a potent, thrombin-dependent stimulation of prothrombinase activity was measured in platelet suspensions that were incubated with a fibrin clot. These results indicate that, in the absence of coagulating plasma, thrombin is a moderate inducer of the procoagulant response of platelets, once integrin alpha(IIb)beta(3)-mediated interactions are stimulated (by adrenaline) and CaCl(2) is present. PMID- 10461012 TI - Elevated thrombotic activity after myocardial infarction: A 2-year follow-up study. AB - This study examines the evolution of the thrombotic activity in patients with myocardial infarction (MI) treated with aspirin (200 mg/day) for 2 years after MI. Plasma samples of 10 patients were collected at 7, 30, 60, 90, 120, 150, 180, 360 and 720 days. In all the samples we measured fibrinogen (Fg), high molecular weight Fg (HMW-Fg), fibrinopeptide A (FPA), prothrombin fragment 1+2 (F1+2), beta thromboglobulin (beta-TG), von Willebrand factor (vWF), tissue factor (TF) and TF pathway inhibitor (TFPI). The plasma Fg, HMW-Fg, FPA, F1+2, beta-TG and vWF levels were significantly elevated in the patients at the beginning of the study as compared to the normal group. The 95% confidence intervals were Fg 277-333 mg/dl, HMW-Fg 200-244 mg/dl, FPA 5.3-16.5 ng/ml, F1+2 1.4-1.8 nmol/l, beta-TG 110 118 IU/ml and vWF 139-195%. At thirty days Fg and HMW-Fg returned to normal levels, whereas the increase in FPA and F1+2 levels persisted throughout the study. At 120 and 150 days, respectively, beta-TG and vWF returned to normal levels. The increase in thrombin generation and activity pointed to a persistent hypercoagulable state 2 years after MI. Plasma levels of TF and TFPI showed no statistically significant variations with respect to the normal values over the 2 year period studied. In conclusion, these results suggest a persistent generation and activity of thrombin and cellular activation in these patients after MI. PMID- 10461013 TI - Glycemic control and coagulation inhibitors in diabetic patients. AB - OBJECTIVE: To investigate the plasma antigenic levels and functional activities of coagulation inhibitors in poorly controlled diabetic patients and the possible effect of good glycemic control on these parameters. RESEARCH DESIGN AND METHODS: Both functional activities and plasma antigenic levels of coagulation inhibitors (antithrombin III, heparin cofactor II, protein C, and protein S) and plasma levels of C4b-binding protein were measured in 28 diabetic patients (13 males, 15 females; 2 IDDM, 26 NIDDM; median age 56.5 years; median duration of diabetes 5.5 years) with poor glycemic control (median HbA(1c) 11.8%). Twenty-three healthy subjects were enrolled as controls. Following a 3-month intensification of antihyperglycemic therapy, good glycemic control (HbA(1c) <8%) was achieved in 17 patients, and the plasma levels of the same parameters during this period were compared with baseline values. RESULTS: Functional activities and plasma antigenic levels of coagulation inhibitors were comparable in poorly controlled diabetic patients and healthy subjects. In patients achieving good control after 3 months, there was a significant reduction in plasma antigenic levels of protein S (p = 0.005) and C4b-binding protein (p = 0.03); however, no difference could be observed in other parameters. HbA(1c) did not show any correlation with plasma antigenic levels or functional activities of coagulation inhibitors either at baseline or at 3 months of good glycemic control. CONCLUSIONS: Our findings suggest that in poorly controlled diabetic patients, coagulation inhibitors are not different from healthy controls. Short-term good glycemic control may not exert a profound effect on coagulation inhibitors except protein S and its binding protein, C4b-binding protein. PMID- 10461014 TI - The beneficial effect of a beta-D-xyloside, Iliparcil, in the prevention of postthrombolytic rethrombosis in the rat. AB - The effect of Iliparcil, a new orally active beta-D-xyloside venous antithrombotic, was studied on the rethrombosis following thrombolytic therapy in rats, using a modified Umetsu model. The drug was administered by oral route prior to thrombolytic therapy, which consisted of administering a combination of heparin and urokinase (H/U) at 37.5 and 70,000 IU/kg, respectively. Time to reocclusion increased from 3.9 min with saline to 10.5 min following H/U injection. When Iliparcil (30 mg/kg, oral route) was administered 4 h before H/U injection, the time to reocclusion was increased by 250% compared with H/U alone (p < 0.001). Similarly, dermatan sulfate (DS), administered intravenously (3 mg/kg) 5 min before thrombus induction, also increased the time to reocclusion (300% compared with H/U alone; p < 0.001). It was also shown that times to reocclusion following Iliparcil or DS treatments were still increased even when heparin dosage was decreased. These results suggest that an antithrombotic product derived from the beta-D-xyloside family could be advantageously used in combination with thrombolytic treatment instead of heparin, which causes complications and side effects. PMID- 10461015 TI - Comparative study of a portable monitor for prothrombin time determination, Coaguchek, with three systems for control of oral anticoagulant treatment. AB - Self-testing of oral anticoagulation is a new possibility related to the development of portable capillary whole blood prothrombin time monitors. The aim of this study was to evaluate one of this monitors, Coaguchek, with respect to its comparability with our routine prothrombin time determination system, as well as with the reference manual technique and two thromboplastins of high sensitivity, Manchester Reagent and one manufactured in our center, Thromboplastin Bilbao, in a group of patients on oral anticoagulant treatment. Although a correlation of r = 0.9271 was found between international normalized ratio (INR) values of Coaguchek and our routine method, Neoplastine/STA analyzer, the difference of the INR scatter increased with the magnitude of measurement, being lowest for INR between the portable monitor and Manchester Reagent and Thromboplastin Bilbao, with a similar coefficient of correlation, r = 0.8948 and r = 0.8905, respectively. A test was performed showing a 65.6% agreement with the INR values of the STA analyzer, 66.4% with Manchester Reagent and 73.4% with Thromboplastin Bilbao. On the basis of this correspondence with laboratory prothrombin time results Coaguchek may be considered as a possible option for monitoring anticoagulated patients even though patients should be given instructions and advice as regards the management and interpretation of the results. PMID- 10461016 TI - Genetic determinants of bone mass. AB - A genetic contribution to bone mass determination was first described in the early 70s. Elucidation of gene contribution to this has since been attempted through studies analyzing associations between bone mass acquisition and/or maintenance and polymorphic variations of several genes. The first to be described was the vitamin D receptor gene (VDR), initially claimed to contribute to almost 75% of the genetic variation in bone mineral density (BMD) in twin and general population studies. Not all of the studies published to date conclude that a clear relationship exists between polymorphic VDR alleles and BMD, and the molecular basis for the VDR gene polymorphisms influence on bone mineralization has not yet been clarified. Since then, other genes with a significant role in bone metabolism such as estradiol receptor, collagen type 1alpha1, TGF-beta1, interleukin-6, calcitonin receptor, alpha2-HS-glycoprotein, osteocalcin, calcium sensing receptor, interleukin-1 receptor antagonist, beta3-adrenergic receptor, apolipoprotein E, PTH, IGF-I and glucocorticoid receptor have been analyzed. Some polymorphic variations in these genes have been associated in some works with significant differences in BMD, with even more significant contributions when associations of different gene polymorphisms were analyzed. Again, the molecular basis for the contribution of these alleles to bone mass determination has not yet been described. A different approach has been attempted by linkage analysis of loci involved in bone density in pedigrees with low BMD using BMD as a quantitative trait. Recent results do not confirm, in these families, any association with any of the previously reported genes, but rather with other as yet unidentified genes. The genetic contribution to mild variations in the general population, as a result of environmental and endogenous individual influences, probably differs completely from that providing a pathologic BMD. PMID- 10461017 TI - Prevalence of coeliac disease in patients with thyroid autoimmunity. AB - The occurrence of autoimmune thyroid disorders among patients with coeliac disease (CD) is well documented, but the exact prevalence of CD among patients with autoimmune thyroid diseases (ATD) is as yet unclear. We screened 150 newly diagnosed patients with ATD by serum endomysial antibody detection (EmA). In 5 subjects (3.3%) EmA positivity was found; all underwent jejunal biopsy. On gluten free diet an excellent clinical and histological response was recorded with an improvement of hypothyroidism and reduction of the thyroxine dosage. Our data suggest a significant high prevalence (3.3%) of CD in patients with ATD, in particular with Hashimoto's thyroiditis. PMID- 10461018 TI - Long-term treatment of growth hormone insensitivity syndrome with IGF-I. Results of the European Multicentre Study. The Working Group on Growth Hormone Insensitivity Syndromes. AB - A total of 33 patients (17 female, 16 male) with Laron syndrome (n = 31) or hGH-1 gene (n = 2, type IA deletion) from 22 centres in 12 countries were enrolled in a study conducted by Pharmacia & Upjohn, Stockholm, which was designed to test the efficacy, in terms of growth promotion and safety, of IGF-I (Igef(TM)). The patients were treated with 40-120 microg/kg IGF-I s.c. twice daily after meals. After the study ended, the patients continued to be treated on an individual basis. The results of 17 patients, who were treated for 48 months or longer were available for the present analysis. Six patients were treated for up to 72 months. When treatment started, the mean age of these patients (8 female, 9 male) was 9.1 (3.7-13.5) years and mean height was -6.5 +/- 1.3 SDS. At the end of the observation period, the mean age of the 17 patients was 14.2 (9.1-17. 7) years and mean height was -4.9 +/- 1.9 SDS. All patients showed a significant increase in growth during the final year on IGF-I, with two of them reaching the age corresponding 3rd centile. The total gain in height (DeltaHT) was 1.7 +/- 1.2 SDS. DeltaHT SDS correlated negatively with age at onset of treatment (R(2) = 0.78, p < 0.02). BMI was 0.6 +/- 1.8 SDS at start of treatment and 1.8 +/- 1.5 SDS at the end of observation. Total DeltaHT SDS correlated positively with total DeltaBMI SDS (R(2) = 0.59, p < 0.01). Long-term treatment of patients with GHIS thus proved to be effective in promoting growth. If treatment is started at an early age, there is considerable potential for achieving height normalisation. The treatment modalities need to be optimized with respect to the growth promoting and metabolic effects of IFG-I. PMID- 10461019 TI - A novel frameshift mutation in the first exon of the 21-OH gene found in homozygosity in an apparently nonconsanguineous family. AB - Congenital adrenal hyperplasia is most frequently due to steroid 21-hydroxylase (21-OH) deficiency. Due to the existence of a pseudogene in tandem duplicated with the 21-OH gene, asymmetric recombination causes the majority of the molecular defects underlying this deficiency: gene conversions and deletions of the functional gene. Screening for a small array of mutations, those existing in the pseudogene together with deletions, allows the characterization of most mutated alleles, 91% in the Spanish population. We report the case of a boy from a nonconsanguineous family, diagnosed during the neonatal period of a salt wasting form of the deficiency, in which this screening did not allow the characterization of the paternal or the maternal allele. This infrequent finding in a nonconsanguineous family was further investigated. Single-strand conformation polymorphism screening for new mutations revealed an abnormally migrating pattern when polymerase chain reaction fragments from 21-OH gene exon 1 of the patient and relatives were analyzed. Upon direct sequencing, the insertion of a T at position 64 (64insT, frameshift generating a stop codon at exon 2) was found in homozygosity in the patient. Microsatellite typing in the HLA region revealed the patient to be homozygous for five markers (heterozygosities 0.62 to 0.74). Apparently this new mutation was generated several generations ago and has been preserved for years. Consanguinity had been discarded for several generations, although both families could be traced back to a small rural area in Navarra (Spain). PMID- 10461020 TI - Clinical usefulness of TSAb assay with high polyethylene glycol concentrations. AB - We previously demonstrated the stimulatory effect of polyethylene glycol (PEG) on thyroid-stimulating antibody (TSAb)-IgG-stimulated cAMP production (thyroid stimulating (TS) index) in porcine thyroid cell (PTC) assay. In the present study the clinical usefulness of the practical method using high PEG concentrations was examined. TS activity using PEG 22.5% precipitated fraction (PF) was significantly higher compared to standard TSAb activity using 12.5% PF from TSAb positive serum, but the maximum TS activity was observed with PEG 12.5% PF + 4% PEG or PEG 22.5% PF + 2% PEG. In all cases of untreated Graves' patients, TSAb activity determined by PEG 22.5% PF was higher compared to standard TSAb activity using PEG 12. 5% PF from test serum, but the highest TSAb activity was observed by PEG 12.5% PF + 4% PEG without increased cAMP production to normal serum. TSAb was positive in 85% (40/47), 98% (46/47) and 100% (47/47) of untreated Graves' patients by the method of PEG 12.5% PF, PEG 22.5% PF and PEG 12.5% + 4% PEG, respectively. Increased TSAb activity by PEG 12.5% PF + 4% PEG method was also observed even if the standard TSAb activity using PEG 12.5% PF method was negative in the euthyroid states of Graves' patients during antithyroid drug therapy. The stimulatory effect of PEG on TS activity was not found in other thyroidal diseases [thyroiditis chronica (with high serum TSH), thyroid stimulation-blocking antibody (TSBAb)-positive sera (with low serum TSH), adenomatous goiter, subacute thyroiditis, and thyroid cancer]. The stimulatory effect of 5% PEG on TS activity produced directly by small amounts of Graves' serum (50 microl) was also found, although the sensitivity was lower than with PEG-precipitated IgG from 0.2 ml serum. The clinical usefulness of the sensitive TSAb assay using PEG-precipitated IgG or direct serum assay in the presence of high PEG concentrations was demonstrated. PMID- 10461021 TI - Vitamin-D receptor genotype does not predict bone mineral density, bone turnover, and growth in prepubertal children. AB - We examined whether the polymorphism for BsmI restriction enzyme in the vitamin-D receptor (VDR) gene influenced radial (distal third) and lumbar (L2-L4) bone mineral density (BMD), phospho-calcium metabolism (calcium, phosphate, intact parathyroid hormone, 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D), biochemical markers of bone formation (osteocalcin and carboxy-terminal propeptide of type-I procollagen) and bone resorption (carboxy-terminal telopeptide of type-I collagen and urinary cross-linked N-telopeptides of type I collagen), insulin-like growth factor I and insulin-like growth factor-binding protein 3, and growth in 209 healthy prepubertal children (112 males and 97 females) aged 7.1-10.0 years. Genotype frequencies were BB 19%, Bb 46%, and bb 35% in the pooled group of children. Clinical findings, dietary calcium intake, calcium density, and physical activity rate were not different (p NS) among the VDR genotypes. Radial BMD, lumbar BMDarea and lumbar BMD adjusted for the apparent bone volume (BMDvolume), and all the biochemical parameters did not differ (p NS) in relation to the VDR genotype. In conclusion, our data show that polymorphism for BsmI restriction enzyme in the VDR gene is not associated with radial and lumbar BMD, parameters of phospho-calcium metabolism and bone turnover, growth hormone-dependent growth factors, and growth in prepubertal children. PMID- 10461023 TI - Activation of arcuate nucleus neurons by systemic administration of leptin and growth hormone-releasing peptide-6 in normal and fasted rats. AB - Both leptin and growth hormone secretagogues are believed to have stimulatory effects on the hypothalamic growth hormone pulse generator, though whether these are achieved through the same pathway is unknown. Systemic administration of a normally maximal effective dose of the growth hormone secretagogue GHRP-6 to male rats causes the induction of c-Fos protein in the ventromedial aspect of the hypothalamic arcuate nucleus. The effect of the same dose of GHRP-6 is, however, much greater in animals that have been fasted for 48 h, suggesting that in the food-replete rat, arcuate neurons either show reduced sensitivity to endogenous growth hormone secretagogues or they are under the tonic inhibitory influences of other factors. The major populations of arcuate neurons activated by GHRP-6 have been shown to contain neuropeptide Y or growth hormone-releasing factor, while leptin is thought to be inhibitory to neuropeptide Y neurons. Leptin did not alter the response of the rats to GHRP-6. However, it was able by itself to induce c-Fos protein immunoreactivity in the ventral, including the ventrolateral, arcuate nucleus of fasted rats. This is a clear demonstration of the acute activation of arcuate neurons in the rat following systemic leptin injection and suggests that GHRP-6 and leptin act on the growth hormone axis via different pathways. PMID- 10461022 TI - Chronic central infusion of growth hormone secretagogues: effects on fos expression and peptide gene expression in the rat arcuate nucleus. AB - Growth hormone (GH) secretagogues induce GH release, in part, by direct actions upon anterior pituitary somatotropes and, in part, by actions upon the neuroendocrine circuitry that regulates GH secretion. In particular, acute systemic administration of GH secretagogues results in increased neuronal activity and Fos protein expression in the arcuate nucleus of the hypothalamus. Prolonged administration of GH secretagogues has been reported to have long lasting effects upon GH release, promoting increased pulsatile secretion. Here, we investigated how chronic central infusion of GH secretagogues affects the response of arcuate nucleus neurons to acute systemic administration of GH secretagogues. In male rats, after central infusion of GH secretagogues for 5 days, there was no sustained expression of Fos in the arcuate nucleus, no significant induction of Fos expression in response to acute GH secretagogue challenge, and a greatly attenuated secretion of GH in response to acute GH secretagogue challenge, all reflecting loss of funtional responsiveness to GH secretagogues. In situ hybridisation revealed that, in the arcuate nucleus of GH secretagogue-infused rats, mRNA levels for GH-releasing hormone, neuropeptide Y and somatostatin were not different than in saline-infused animals. However, somatostatin mRNA levels in the periventricular nuclei of GH secretagogue-infused rats were significantly higher than those of saline-infused rats, indicating that this nucleus may play an important role in mediating the effects of chronic GH secretagogue administration. PMID- 10461024 TI - Growth hormone-releasing hormone and morphine attenuate growth hormone secretagogue-induced activation of the arcuate nucleus in the male rat. AB - Growth hormone secretagogues (GHS) administered systemically selectively induce growth hormone (GH) release from the pituitary and the expression of Fos protein in arcuate nucleus neurons. Both the control of GH release and the expression of the GHS receptor in the arcuate nucleus are thought to be regulated, at least in part, by the negative feedback actions of GH. In this study, we utilized the immunocytochemical detection of Fos protein to examine the effects of morphine- and GH-releasing hormone (GHRH)-induced GH release on the activation of arcuate nucleus neurons following GHS administration. Given alone, two structurally different GHS induced significant amounts of Fos-LI in the arcuate nucleus of male rats, suggesting activation of cells in this region. Prior administration of morphine or GHRH significantly reduced the number of Fos-positive cells in the arcuate nucleus of rats injected with either GHS, although when given together, morphine and GHRH did not produce a greater reduction in Fos expression than when given alone. In no case was there a complete reduction in Fos expression, indicating that some arcuate nucleus neurons are not subject to the feedback effects of endogenous GH. These results provide evidence that, in the male rat, GH can feedback to the hypothalamus, altering the responsiveness of neurons involved in the central response to GHS. PMID- 10461025 TI - Transgenic mice overexpressing the growth-hormone-releasing hormone gene have high concentrations of tachykinins in the anterior pituitary gland. AB - According to recent reports, substance P (SP) is localized in the anterior pituitary gland within subsets of thyrotropes and somatotropes, although earlier electron-microscopic studies described the presence of this tachykinin in mammotropes and gonadotropes. Transgenic mice overexpressing the growth-hormone releasing hormone (GHRH) gene have markedly enlarged pituitary glands, due to hyperstimulation of the somatotropes. Therefore, we speculated that if somatotropes are able to synthesize tachykinins, these peptides should be greatly increased in the anterior pituitary of transgenic GHRH mice. We found that, in accordance with our working hypothesis, both SP and neurokinin A (NKA) were markedly increased in the anterior pituitary gland of male and female transgenic mice, compared with their respective normal controls. In male transgenic mice, NKA was 13.6- and SP 20.2-fold higher than in the anterior pituitary from normal mice. In female transgenic mice, NKA was 40- and SP 100-fold higher than in the anterior pituitary from normal female mice. In male transgenic mice, NKA and neuropeptide K (NPK) contents in the anterior pituitary showed no significant changes between 26 and 50 days of age but significantly increased between 50 days and 5 months of age. The concentration of NKA in the anterior pituitary did not show significant differences between 26 days and 5 months of age, but NPK concentrations in the anterior pituitary significantly decreased with age. In female transgenic mice, NKA content and concentration in the anterior pituitary increased after 35 days of age, but NPK concentrations significantly decreased after 26 days of age. Triiodothyronine markedly decreased anterior pituitary tachykinins, but ovariectomy and estrogen administration failed to significantly affect tachykinin concentrations in the anterior pituitary of transgenic mice. Tachykinin immunostaining was detected in some somatotropes, but tachykinins were also present in cells that were not GH positive. These findings indicate that hyperstimulated somatotropes contain increased stores of tachykinins and that these cells are a source of tachykinins in the anterior pituitary. Tachykinin stores in the anterior pituitary of transgenic mice were affected by thyroid hormones but seem to be insensitive to estrogens. The GHRH transgenic mice may be an interesting model to study the regulation of tachykinin stores in the anterior pituitary gland. PMID- 10461026 TI - Development of a site-directed polyclonal antibody against the pituitary growth hormone-releasing hormone receptor and its use to estimate GHRH receptor concentration in normal and hypothyroid rats. AB - A site-directed polyclonal antipeptide antibody was generated in rabbits against segment 392-404 of the rat pituitary growth hormone-releasing hormone receptor (GHRH-R), using a multiple antigenic peptide system strategy of immunization. This C-terminal intracellular region of the rat GHRH-R exhibits 85% sequence identity with the human GHRH-R. The purified anti-GHRH-R(392-404) IgGs were characterized in cell lines expressing the human GHRH-R and in rat and human anterior pituitary, using immunoblotting. The polyclonal antibody recognized a 45 kD protein in human GHRH-R-transfected BHK 570 cell membrane preparations but not in wild-type cells. A 45-kD N(alpha)-tagged human GHRH-R was immunodetected with both antitag and anti-GHRH-R antibodies in human GHRH-R-transfected HEK 293 cells. Cross-linking of [(125)I-Tyr(10)]hGHRH(1-44)NH(2) to GHRH-R-transfected BHK cells led to the detection of a major and specific 45-kD radioactive complex. Its probing with the anti-GHRH-R(392-404) IgGs led also to the detection of a 45 kD entity. In rat anterior pituitary homogenates or membrane preparations, immunoblotting led to the detection of 44-, 47- and 65-kD proteins. In human anterior pituitary membrane preparations, immunoblotting led to the detection of 52- and 55-kD proteins. No immunoreactive signal was observed in the rat liver. Cross-linking of [(125)I-Tyr(10)]hGHRH(1-44)NH(2) to rat anterior pituitary homogenates revealed the presence of specific 28-, 47- and 65-kD radioactive complexes. Probing of these radioactive complexes with the anti-GHRH-R(392-404) IgGs resulted in the visualization of 28-, 47- and 65-kD entities and of an additional immunoreactive 44-kD protein. To assess the usefulness of this GHRH-R antibody, estimation of changes in the concentration of rat anterior pituitary GHRH-R was performed by immunoblotting and compared to binding data after a 3 week antithyroid treatment. The treatment known to depress the 2.5- and 4-kb GHRH R mRNA transcripts by at least 1.7-fold decreased the apparent maximal concentration of high (B(max1)) and low (B(max2)) affinity binding sites by 4.6- and 15.2-fold, respectively, and the 47- and 65-kD GHRH-R proteins by 3.5- and 1. 25-fold, respectively. Altogether, the characteristics of the anti-GHRH-R(392 404) polyclonal antibody indicate that it specifically recognizes the human and rat GHRH-R. It also represents an additional valuable tool to estimate variations of GHRH-Rs in physiopathological conditions known to affect GHRH-R mRNA and/or GHRH binding site concentrations. PMID- 10461027 TI - Restricted presence of the growth hormone-releasing hormone receptor to somatotropes in rat and human pituitaries. AB - Specific binding of growth hormone-releasing hormone (GHRH) to its plasma membrane receptor represents the first step of cellular signals leading to exocytotic GH secretion in the anterior pituitary. The GHRH receptor (GHRH-R) has been cloned and belongs to the secretin/glucagon/vasoactive intestinal peptide subfamilly of G-protein-coupled receptors. To study its characteristics in rat and human pituitaries and examine its cellular and subcellular localization, a site-directed polyclonal antibody recognizing the C-terminal portion 392-404 of the rat and human GHRH-R was used. Immunohistochemistry was performed on paraffin embedded pituitary sections while ultrastructural immunocytology was done on frozen and Lowicryl-resin-embedded ultrathin sections. GHRH-R-like immunoreactivity was restricted to somatotropes and colocalized with GH in both rat and human tissues. No signal was detected in gonadotropes, lactotropes, corticotropes and thyrotropes. At the subcellular level, gold particles were associated with the plasma membrane (observed on ultrathin frozen sections), secretory granule membrane, cytoplasmic matrix, nuclear membrane and nuclear matrix. In the nucleus, gold particles were mainly observed at the junction between eu- and heterochromatin. The highest density of labeling was observed in the cytoplasm (55 vs. 45% in the nucleus), mainly in secretory granules (59% of cytoplasmic labeling) and the plasma membrane. These results support the hypothesis that GHRH-mediated actions in the pituitary are specific to somatotropes and that GHRH-R isoforms and/or ligand-receptor complexes are involved in intracellular trafficking, recycling processes and nuclear functions. PMID- 10461028 TI - Identification of endocannabinoids and cannabinoid CB(1) receptor mRNA in the pituitary gland. AB - Most data on effects of natural and synthetic cannabinoids on anterior pituitary hormone secretion point out to a primary impact on the hypothalamus. There is also some evidence, however, of possible direct actions of these compounds on the anterior pituitary, although the presence of cannabinoid receptors in the pituitary has not been documented as yet. In the present study, we evaluated the presence of cannabinoid CB(1) receptor-mRNA transcripts in the pituitary gland by in situ hybridization. We observed CB(1) receptor-mRNA transcripts in the anterior pituitary and to a lesser extent in the intermediate lobe whereas they were absent in the neural lobe. We then examined whether CB(1) receptor-mRNA levels in both pituitary lobes responded to chronic activation by a specific agonist, as did receptors located in adjacent hypothalamic nuclei and in other brain regions. Daily administration of CP-55,940 for 18 days produced a small, but statistically significant paradoxical increase in CB(1) receptor-mRNA levels in the anterior pituitary, with no changes in the intermediate lobe, in contrast to reduced CB(1) receptor-mRNA levels observed in the ventromedial hypothalamic nucleus (VMN), and to decreased CB(1) receptor binding in the VMN and the arcuate nucleus. The time-course of up-regulation of CB(1) receptor-mRNA transcripts in the anterior lobe was biphasic; daily administration of Delta(9) tetrahydrocannabinol produced an early and marked decrease in CB(1) receptor-mRNA levels after 1 and 3 days, followed by normalization after 7 days and by a small increase after 14 days. We also checked whether endogenous cannabinoid ligands are present in the anterior pituitary and the hypothalamus. Although anandamide itself was detected only in trace amounts, concentrations of its precursor N arachidonoyl-phosphatidyl-ethanolamine and of 2-arachidonoyl-glycerol were found in both tissues, suggesting that endocannabinoids may be synthetized in the anterior pituitary. In summary, CB(1) receptors and corresponding ligands seem to be expressed in cells of the anterior and intermediate lobes of the pituitary, but the response of CB(1) receptor-mRNA transcripts in the anterior lobe to chronic agonist activation is different than the desensitization observed in hypothalamic nuclei. PMID- 10461030 TI - IgA nephropathy in children. PMID- 10461029 TI - Insulin metabolism in Alzheimer's disease differs according to apolipoprotein E genotype and gender. AB - Higher fasting plasma insulin levels and reduced CSF-to-plasma insulin ratios, suggestive of insulin resistance, have been observed in patients with Alzheimer's disease (AD) who do not possess an apolipoprotein E (APOE)-epsilon4 allele. We examined the relationship of APOE and gender to peripheral insulin action and hyperinsulinemic memory facilitation in patients with AD using a sensitive measure of insulin-mediated glucose disposal. Participants were 32 patients with AD (9 without an epsilon4 allele, 23 with an epsilon4 allele) and 25 healthy age matched adults (16 without an epsilon4 allele, 9 with an epsilon4 allele). AD subjects without an epsilon4 allele had significantly lower insulin-mediated glucose disposal rates than AD patients with an epsilon4 allele (p < 0.03), or than normal adults without an epsilon4 allele (p < 0.02). Female AD subjects showed lower insulin-mediated glucose disposal rates than did male AD subjects (p < 0.02). No significant interaction was observed between APOE group and gender, suggesting that these effects are independent. AD subjects without an epsilon4 allele also showed significant memory facilitation in the hyperinsulinemic condition (p < 0.04), whereas the AD-epsilon4 group did not. Also in the hyperinsulinemic condition, AD patients without an epsilon4 allele had lower insulin levels than patients with an epsilon4 allele (p < 0.02), and women with AD had lower insulin levels than did men with AD despite similar insulin infusion rates and body mass (p < 0.004). No gender or genotype effects were observed in either condition for normal subjects. These results provide in vivo evidence of differences in insulin-mediated energy metabolism between epsilon4 and non epsilon4 AD, and suggest that defective insulin action may be of particular pathophysiologic significance for patients without an epsilon-4 allele. PMID- 10461031 TI - Chronic rejection in kidney transplants. An in-depth review. PMID- 10461032 TI - Increased prevalence of apolipoprotein E3/E4 genotype among Swedish renal transplant recipients. AB - There is increasing evidence that lipoproteins are involved in the progression of kidney diseases and in the deterioration of kidney transplant function, although the exact mechanism is still not known. Common polymorphisms of apolipoprotein E genotype associate with the variability of lipoprotein levels and composition. We have, therefore, determined the apolipoprotein E genotype in a group of 112 renal transplant patients, of whom 27 had had an episode of acute vascular rejection, while 85 had not. We found no difference in apolipoprotein E genotype distribution or in relative allele frequency in the vascular rejection group as compared with the group without vascular rejection. The apolipoprotein E genotype distribution in the transplant group was also compared with that in a group of 407 healthy Swedish individuals. The E3/E4 genotype occurred with a significantly increased frequency in the transplant group: 38.3 versus 16% in the control group (p < 0.001). The prevalence of individuals carrying the epsilon4 allele among the transplant group was also significantly higher (44%) as compared with the control group (30%; p < 0.01). This increase was entirely due to the predominant increase of E3/E4, as the E4/E4 genotype was less frequent in transplant recipients than in normal controls (3.5 vs. 10.6%; p < 0.05). The relative frequencies of epsilon2 (0.044), epsilon3 (0.716), and epsilon4 (0.238) alleles in the renal transplant group were not different from those of normal controls (0. 078, 0.718, and 0.202, respectively). With regard to the prevalence of E4/E4 in the two groups, the lack of difference in the relative frequency of the epsilon4 allele must be interpreted with caution. The results thus suggest that the E3/E4 genotype may be associated with the progression of kidney disease leading to renal insufficiency. However, the apolipoprotein E genotype does not seem to influence the risk of vascular rejection among transplant recipients. PMID- 10461033 TI - Quality of life in end-stage renal disease patients after successful kidney transplantation: development of the ESRD symptom checklist - transplantation module. AB - The End-Stage Renal Disease Symptom Checklist - Transplantation Module (ESRD SCL((R))) was developed to assess the specific physical and psychological quality of life of renal transplant recipients, with a special focus on side effects of immune system suppression therapy. A list of potentially relevant items was administered to 458 transplant recipients. The symptoms present in >20% of patients were chosen, and factor analysis was used to create the final questionnaire which consists of 43 items in six dimensions: (1) limited physical capacity (10 items; internal consistency: Cronbach's alpha = 0.85); (2) limited cognitive capacity (8 items, alpha = 0.82); (3) cardiac and renal dysfunction (7 items, alpha = 0. 76); (4) side effects of corticosteroids (5 items, alpha = 0.77); (5) increased growth of gum and hair (5 items, alpha = 0.78), and (6) transplantation-associated psychological distress (8 items, alpha = 0.80). All questions are scored on a five-point Likert scale. Validity was demonstrated in correlation with corresponding SF-36 scales and in a stepwise hierarchical regression model predicting the subscales of the ESRD-SCL by sociodemographic and medical data. The ESRD-SCL was found to have adequate reliability, test-retest correlations in a subsample of 88 stable patients after 1 year, and construct validity. PMID- 10461034 TI - Suppressive effects of Perilla frutescens on spontaneous IgA nephropathy in ddY mice. AB - Perilla frutescens (perilla) is a common herb used in Japan for garnishing raw seafood to protect the alimentary tract from inflammatory diseases. The present study was performed to investigate whether or not perilla prevents the development of lesions of IgA nephropathy in ddY mice which spontaneously develop this disease. After orally administering perilla extract to ddY mice from 8 to 42 weeks of age, the changes in urine, serum, and kidneys were evaluated. Perilla extract significantly suppressed proteinuria and glomerular IgA deposition (p < 0.01 and p < 0.05, respectively). The decreased serum IgA concentration in perilla-treated mice showed a significant correlation with glomerular IgA deposition. Such findings suggest that perilla reduced glomerular IgA deposition via suppression of IgA production in the serum. On the other hand, the nitric oxide concentration in the serum of perilla-treated mice was significantly higher than that observed in the controls. The addition of the sera of perilla-treated mice to quiescent cultured murine mesangial cells resulted in a cell proliferation which was less than in controls, suggesting that perilla might either directly prevent mesangial cell proliferation or prevent proliferation by regulating circulating cytokines. Such results indicate that perilla should prevent IgA nephropathy, thus representing a promising herbal medicine for glomerulonephritis. PMID- 10461035 TI - Anticardiolipin antibodies in acute poststreptococcal glomerulonephritis and streptococcal impetigo. AB - Anticardiolipin (aCL) antibodies have been described in diverse clinical situations, linked to the risk of thrombosis in different vascular locations. They have been rarely studied in renal diseases, and occasionally they have been associated with glomerular thrombosis. We analyzed the incidence of aCL (isotypes IgG, IgA, and IgM) in samples, taken during the acute phase of the disease, from 27 well-documented patients having acute poststreptococcal glomerulonephritis. Twelve cases were positive on IgG testing, 1 case on IgA testing only, and no one was positive on IgM testing. A serological follow-up was performed with a second sample taken about 7 months later, for the patients initially positive on IgG testing showing persistence in 9. Clinical variables during the acute phase and after a follow-up period of 25 (range 6-89) months were analyzed for possible associations with the presence of these antibodies, but non was significantly related. Renal histopathological investigation did not reveal particular findings in the aCL-positive patients, and glomerular thrombosis was not found in any case. In addition, serum samples from 12 streptococcal impetigo patients without renal involvement were analyzed, showing similar incidence (4 positive on IgG testing, 1 of them positive on IgM testing as well, and no one positive on IgA testing) and titers of aCL antibodies. We conclude that the presence of aCL antibodies in acute poststreptococcal glomerulonephritis may be a marginal immunological phenomenon unrelated to the glomerular disease, triggered by the streptococcal infection. PMID- 10461036 TI - Initial ultrastructural changes in pore size and anionic sites of the glomerular basement membrane in streptozotocin-induced diabetic rats and their prevention by insulin treatment. AB - BACKGROUND/AIM: The present study was conducted to elucidate the mechanism(s) of the development of early diabetic nephropathy, examining ultrastructural changes employing electron microscopy, especially changes in pore size of the glomerular basement membrane (GBM) of streptozotocin (STZ)-induced diabetics rats. METHODS: Urinary albumin excretion rate (UAE), pore size of the lamina densa of the GBM visualized directly by the tissue negative staining method, and number of anionic sites (AS) in the corresponding portion of the lamina rara externa were determined for 6 weeks in diabetic rats without and with insulin treatment. RESULTS: The UAE of the diabetic rats increased with time and was significantly greater than that of the nondiabetic control rats after 4 weeks (p < 0.01), while insulin treatment suppressed the increased UAE of diabetic rats. The median values in both short diameter and long dimension of the pores in the diabetic group were markedly increased at the 2nd week as compared with those in the nondiabetic control rats, whereas no significant change was found in the pore size of the diabetic rats with insulin treatment. Moreover, the number of AS in the GBM of the diabetic rats was significantly (p < 0.001) decreased from the 2nd week onward. Insulin treatment also prevented a decrease in AS number in diabetic rats. CONCLUSIONS: It is suggested from these results that an impairment of barrier size selectivity occurs at a very early stage of STZ-induced diabetes in rats, which may enhance the abnormality of the charge-selective properties of the GBM. In addition, insulin treatment may protect this barrier system through normalizing blood glucose control in STZ-diabetic rats. PMID- 10461037 TI - Influence of parathyroidectomy and calcium on rat renal function. AB - Parathyroid hormone (PTH) has multiple effects on water and electrolyte transport along the nephron. However, the influences of PTH and calcium on the urinary concentration ability are not fully understood. In this study, clearance and microperfusion studies were performed in thyroparathyroidectomized (TPTX) rats either supplemented (TPTX+Ca(2+)) or not with calcium added to the ingested food as CaCl(2) (1.6 g/100 g). Acid-base data and renal functional parameters were measured in TPTX and TPTX+Ca(2+) rats. Additional studies were performed in the isolated inner medullary collecting tubules of intact and TPTX rats to evaluate the osmotic permeability of this segment in the presence of 10(-6) M PTH added to the bath. In these experiments the possible influence of PTH on antidiuretic hormone induced changes of the osmotic permeability in TPTX and TPTX+Ca(2+) rats was also investigated. In the TPTX+Ca(+) group, the glomerular filtration rate increased significantly when compared to the TPTX group (6.04 +/- 0.42 vs. 4.88 +/- 0.20 ml.min(-1).kg(-1); p < 0.05), but the U/P inulin ratio remained lower than control values (30.8 +/- 1.48 vs. 54.0 +/- 3.5; p < 0.05), which suggests that normal levels of PTH are necessary to maintain the concentrating ability. In a group of TPTX rats, an acute infusion of PTH (0.5 microg.min(-1).kg(-1)) significantly decreased the urinary flow and increased the renal plasma flow, results that agree with the vasomodulator action of this hormone on the renal vasculature. A significant increase in the fractional K(+) excretion observed in the TPTX+Ca(2+) group as compared with both control and TPTX, groups suggests that the excreted load of Ca(2+) may interfere with tubular K(+) handling in the absence of PTH. PTH (10(-6) M) added to the bath of the isolated inner medullary collecting tubules did not change the osmotic permeability, of intact, TPTX, and TPTX+Ca(2+) rats. Furthermore, it did not modify the antidiuretic hormone induced changes in the osmotic permeability. These results suggest that this segment of the nephron is PTH insensitive as far as water and ion transport are concerned. PMID- 10461038 TI - Enhanced hyaluronan synthesis in the MRL-Fas(lpr) kidney: role of cytokines. AB - BACKGROUND/AIM: Pathological accumulation of the extracellular matrix component hyaluronan (HA) occurs in the kidney cortex in immune-system mediated tissue injury. The purpose of the present study was to examine the pattern of HA deposition and the mechanisms of HA synthesis in the MRL-Fas(lpr) mouse model of lupus nephritis. METHODS: Kidneys from normal and autoimmune mice were examined for HA content by immunofluorescence staining. Steady state mRNA levels for key enzymes involved in HA synthesis - uridine diphosphate-glucose dehydrogenase (UDPGDH) and HA synthases (HAS) 1, 2 and 3 - were assessed by reverse transcriptase polymerase chain reaction (RT-PCR). Using cultured mouse tubular epithelial cells, the regulation of the HA production in vitro in response to tumor necrosis factor alpha and interferon gamma was also examined. RESULTS: By immunofluorescence staining, large amounts of HA were detected in the cortical interstitium of MRL-Fas(lpr) mice with autoimmune renal injury, but not in congenic MRL-++ mice. By RT-PCR the presence of transcripts for several genes involved in the synthesis of HA in normal and autoimmune kidneys could be demonstrated, including mRNA for UDPGDH and HAS1 and HAS2, but not for HAS3. Except for HAS2, steady state mRNA levels for these enzymes did not correlate with disease activity. Analyzing a kidney tubular epithelial cell line in vitro, it was found that tumor necrosis factor alpha and interferon gamma, and particularly the combination of these two cytokines, markedly enhanced the synthesis of HA. The expression of HAS2 mRNA was also enhanced in response to cytokine treatment. CONCLUSIONS: HA deposition is prominent in MRL-Fas(lpr) mice with renal disease and could be mediated by local synthesis through HAS1 and HAS2. We hypothesize that the enhanced synthesis of HA could be promoted by proinflammatory cytokines in vivo. The functional significance of HA accumulation in autoimmune renal injury remains to be determined. PMID- 10461039 TI - Historadioautographic localization, pharmacology and ontogeny of V(1a) vasopressin binding sites in the rat kidney. AB - The localization and pharmacological characteristics of vasopressin (VP) binding sites of the V(1a) subtype in developing and adult rat kidney were investigated by radioautography on kidney sections incubated in the presence of a radioiodinated selective V(1a) antagonist. Their localization after in vivo systemic infusion of the radioligand was also investigated. V(1a) binding sites first appear at embryonic day 16 on vascular elements. In the adult, they were localized in the cortex (vascular and tubular structures, juxtaglomerular apparatus), the outer medulla outer stripe (vasa recta) and inner stripe (thin descending limbs of short looped nephrons) and the inner medulla (collecting ducts). Data obtained in vitro were confirmed by in vivo binding at postnatal day 30 (PN30). Whatever their localizations, the V(1a) binding sites exhibited full V(1a) pharmacological profile in postnatal stages rats and in adult rats: a high affinity (nM range) for VP and for the V(1a) agonist, a lower affinity (microM range) for oxytocin and no affinity for the oxytocin agonist. The presence of V(1a) binding sites in these different structures raises the question of the putative roles of VP in modulating renal functions. A striking finding is the presence of V(1a) binding sites in the outer medullary thin descending limbs of short looped nephrons suggesting their colocalization with urea transporters. PMID- 10461040 TI - Balkan endemic nephropathy. Slowed progression of kidney disease by avoidance of etiological factors. PMID- 10461041 TI - Persistent left superior vena cava detected by hemodialysis catheterization. PMID- 10461042 TI - Complete atrioventricular blockade secondary to conventional-release verapamil in a patient on hemodialysis. PMID- 10461043 TI - Measurement of lean body mass with dual energy X-Ray absorptiometry in dialysis patients and its pitfalls. PMID- 10461044 TI - Losartan in renal allograft recipients receiving cyclosporine A. PMID- 10461045 TI - IgA nephropathy and antiphospholipid syndrome. PMID- 10461046 TI - Antihypertensive medication compliance in African-American stroke patients: behavioral epidemiology and interventions. AB - Hypertension is a major cause of stroke in the African-American community, and lack of control of hypertension appears to be common. Improving compliance to antihypertensive therapy in African-American stroke patients could have a significant impact on recurrent stroke rates. Little is known about factors affecting compliance in this community and which interventions would be effective in improving compliance. Health behavior models which assess the patient's perception of stroke and hypertension, barriers to the desired behavior, perception of ability to perform the behavior, perception of others' acceptance of the behavior and the patient's behavioral stage could be used to tailor interventions to improve compliance. A plan to improve compliance should take into account the target population's baseline rates of compliance, perception of need for intervention and risk factors for noncompliance. PMID- 10461047 TI - Dementia in Argentina and other Latin-American countries: An overview. AB - Latin-American countries are expected to experience an expansion of the elderly population, as life expectancy increases. We reviewed the literature to determine the frequency of dementia in our region and surveyed selected Latin-American countries to determine the availability of diagnostic and treatment services and long-term care facilities. Latin-American countries face a challenge to develop public health strategies to cope with the anticipated heightened number of elderly with cognitive impairment. PMID- 10461048 TI - A report on the WHO working group on Parkinson's disease. AB - In order to raise awareness of the public health importance of Parkinson's disease, the World Health Organisation (WHO) has recently established a Working Group on Parkinson's Disease and included it in the framework of the WHO Global Initiative on Neurology and Public Health. The first meeting of this international expert group produced a set of recommendations covering the following aspects of Parkinson's disease: epidemiology; organisation of services and treatment; education, training and information, and direct and indirect costs of care. An independent international research project entitled Global Parkinson's Disease Survey has recently been launched in response to the recommendations of the WHO Working Group on Parkinson's Disease. This paper summarises the recommendations of this WHO Working Group and outlines objectives, methods and preliminary pilot results of the Global Parkinson's Disease Survey. PMID- 10461049 TI - Greater stroke rate during hospitalization for acute heart disease among Mexican Americans than non-Hispanic whites. AB - BACKGROUND AND PURPOSE: This study compared the risk for stroke during acute myocardial infarction (AMI), percutaneous transluminal coronary angioplasty (PTCA) and coronary artery bypass grafting (CABG) between Mexican Americans (MAs) and non-Hispanic whites. METHODS: We examined the age-specific rate ratios (RR) of acute stroke during hospitalization for AMI, CABG and PTCA in a population based study in Corpus Christi, Tex. by searching the cardiac surveillance data for ICD-9 codes for stroke (430-437). ICD-9 stroke codes were validated by comparing medical chart abstraction with ICD-9 discharge diagnoses. RESULTS: Stroke codes were found in 220 of the 5,697 admissions for AMI, CABG and PTCA. In the 45- to 59-year age-group MAs had a RR of 2.66 (95% CI 1.36-5.23) relative to non-Hispanic whites. In the 60- to 74-year age-group the RR was 1.52 (95% CI 1.11 2.08). There were no significant differences in the 25- to 44-year age-group. These ethnic relationships were found in nondiabetics but not in diabetics. Women in the 45- to 59-year age-group had a RR of 1.88 (95% CI 1.09-3.25) compared with men, but there were no significant sex differences in the 25- to 44- or 59- to 74 year age-groups. Stroke ICD-9 codes have a poor positive predictive value for acute stroke ranging from 10 to 76%. The stroke misclassifications were nondifferential with respect to ethnicity or sex. CONCLUSIONS: MAs have a higher stroke rate complicating acute heart disease in Corpus Christi. A rigorous stroke surveillance project is needed to study the burden of stroke in MAs, the United States' largest Hispanic population. PMID- 10461050 TI - Spectrum of disease in vascular cognitive impairment. AB - The recognition that cognitive impairment of vascular origin is not limited to multi-infarct dementia has led to the development of several sets of new criteria for vascular dementia (VaD). We set out to define the spectrum of disease in patients presenting with vascular cognitive impairment (VCI). Of 412 patients consecutively seen at a memory clinic, 80 had VCI. These patients had vascular cognitive impairment not dementia (n = 19), VaD (n = 48), and mixed Alzheimer's disease-VaD (n = 13). Radiographic patterns were: white matter changes only (40%); multiple infarcts (30%); single strategic stroke (14%), and no identified lesion (16%). Of note, 19 (24%) of these patients meet none of the currently published criteria for VaD. To better understand and treat ischaemic causes of cognitive impairment, the concept of VaD should be expanded to include patients who do not meet traditional dementia criteria. PMID- 10461051 TI - Temporal trend and factors associated with delayed hospital admission of stroke patients. AB - The effectiveness of stroke treatment depends on the time interval between onset of symptoms and admission to hospital. The purpose of our investigation was to assess, over a 10-year period, the mean delay in admission to hospital in stroke patients to determine factors which might be associated with this delay, to define the putative number of patients available for accrual in clinical trials, and to identify strategies aimed at decreasing the time to admission. We collected data on all stroke patients consecutively admitted to our clinic from 1986 to 1995. The following variables were investigated: age, sex, educational and occupational level, home accommodation, family and personal history of vascular disease or factors known to affect the risk of vascular disease, and type and severity of stroke. The individual and independent contribution of these variables was assessed by univariate and multivariate analysis. The accurate time of stroke onset was established for 760 patients. Of these, 24.7% were admitted within 1 h from the onset of symptoms, 41% within 2 h, 54% within 4 h and 72.5% within 12 h. The mean delay was 21 +/- 2 h (SE) and the median was 3.5 h. Acute onset of neurological deficits, stroke severity and family history of cerebrovascular disease were associated with earlier presentation. According to the current guidelines for thrombolytic therapy, only 16% of the patients could have been included in a clinical trial. This study suggests that despite a relatively short time to hospital admission in most patients and an altered help seeking behavior over time, many stroke patients did not present early enough to be recruited for clinical trials or to benefit from new treatments. The majority of patients with timely presentation were not eligible for acute treatment, or were subjects with severe stroke for whom caution is advised before initiating thrombolytic therapy. It has been suggested that the patient's indecision to seek medical help is the most important reason for a delayed hospital admission of stroke patients. These results underscore the importance of interventions aimed at reducing the delay in stroke treatment induced by patients who are unaware of the decisive role of the time of treatment induction. The finding that earliest admissions for stroke comprised patients with a previous history of cerebrovascular disease suggests that an education campaign might highlight the importance of an early admission. PMID- 10461052 TI - Spinal muscular atrophy. Incidence in Iceland. AB - Spinal muscular atrophy (SMA) is among the commonest degenerative disorders of the nervous system in childhood. This is an inherited autosomal recessive disease which results in premature death of anterior horn cells of the spinal cord and is manifested by progressive weakness and atrophy of skeletal muscles. Few studies have looked at the frequency of the disease in a defined population. We identified all patients diagnosed with SMA in Iceland during a 15-year period. The diagnosis is based on typical symptoms and supported by results of electromyography/nerve conduction studies and muscle biopsy. The average annual incidence was 13.7 per 100,000 live births for all types of SMA, which is similar to that reported in other population-based studies. PMID- 10461053 TI - A family history of Parkinson's disease and its effect on other PD risk factors. AB - Parkinson's disease (PD) is likely a result of both inherited and exogenous factors. In a study of 144 PD cases and 464 controls, we used PD family history as a surrogate for inherited PD susceptibility. Cases were more likely to report a first- or second-degree relative with PD: 16.0 vs. 4.3%; odds ratio (OR) = 4. 2; 95% confidence interval (CI) = 2.3-7.6. A PD family history was a greater risk factor for PD in subjects under age 70 (OR = 8.8; 95% CI = 3.4-22.8) compared with those over 70 (OR = 2.8; 95% CI = 1.3-6. 1) and in men (OR = 8.1; 95% CI = 3.4-19.2) compared with women (OR = 2.6; 95% CI = 1.1-6.0). We also tested whether a PD family history modified the effects of other PD risk factors. In subjects with a PD family history, occupational exposure to copper, lead or iron increased the risk for PD (OR = 3.0; 95% CI = 0.7-13.3), but this was not the case for those without a family history (OR = 1.1; 95% CI = 0.7-1.6). Ever smoking cigarettes was inversely associated with PD in those without a PD family history (OR = 0.6; 95% CI = 0.4-0.9), but was positively associated with PD in those with a PD family history (OR = 1.7; 95% CI = 0.5-5.9). In summary, our results suggest that a PD family history, and perhaps, therefore, an inherited susceptibility, confers a greater risk for PD in men and individuals under 70 years of age and may modify the effects of environmental risk factors for PD. PMID- 10461054 TI - Management of non-epithelial ovarian tumours. AB - Ovarian tumours of non-epithelial origin are less common than those of epithelial origin but must be distinguished from these as their natural history and management differ. As these tumours are rare, histological review by an expert in the field is essential. There have been no randomised trials to outline the management of these tumours and therefore this paper represents a review of descriptive data. Non-epithelial ovarian tumours, which represent approximately 50% of all ovarian tumours and approximately 25% of malignant ovarian tumours, are outlined. Lymphomas and metastatic tumours should be treated the same as when found in other sites and will not be discussed further. PMID- 10461055 TI - Induction chemotherapy followed by concurrent chemotherapy and high-dose radiotherapy for locally advanced squamous cell carcinoma of the cervical oesophagus. AB - The efficacy and toxicity of combined radiochemotherapy for locally advanced squamous cell carcinomas of the cervical oesophagus was evaluated retrospectively. Induction chemotherapy consisted of three courses of 5 fluorouracil (5-FU), leucovorin, etoposide and cisplatin (FLEP) or two courses weekly six times of 5-FU and leucovorin combined with biweekly cisplatin. This induction regimen was followed by high-dose external beam radiotherapy up to 60 66 Gy and concurrent chemotherapy with cisplatin and etoposide. Median follow-up of the recruited 17 patients was 37 months (13-73 months). Long-term survival was 24% at 2 and 3 years. The probabilities of locoregional tumour recurrences and distant metastases as sites of first relapse were 67 and 39% at 2 years. Acute and late toxicity of this schedule was moderate. The protocol offers a definitive chance of long-term survival for patients with locally advanced carcinomas of the cervical oesophagus, but local in-field recurrences remain the predominant risk after treatment. Intensification of the regimen seems possible because no dose limiting late toxicities were observed. PMID- 10461056 TI - Efficacy of continuous hyperthermic peritoneal perfusion for the prophylaxis and treatment of peritoneal metastasis of advanced gastric cancer: evaluation by multivariate regression analysis. AB - Thirty-two patients with advanced gastric cancer underwent continuous hyperthermic peritoneal perfusion (CHPP) combined with surgery: to prevent peritoneal recurrence in 15 patients without peritoneal metastasis (prophylactic CHPP) and to treat 17 patients with peritoneal metastases (therapeutic CHPP). The postoperative outcome was compared with that of control patients treated with surgery alone. Peritoneal recurrence was less frequent (26%) and the 5-year survival rate was significantly higher (39%) in the patients with prophylactic CHPP than in 40 control patients (42 and 17%, respectively). The patients with therapeutic CHPP showed significantly better median survival than did 20 control patients (11 vs. 6 months). Cox multivariate regression analysis revealed that CHPP was an independent prognostic factor in the prophylactic study (hazard ratio = 0.3965), and that the independent prognostic factor in the therapeutic study was not CHPP but complete resection of the peritoneal metastasis. Thus, CHPP has no marked benefit for established peritoneal metastasis. CHPP for the prevention of peritoneal recurrence may have a beneficial effect on long-term survival, but a prospective randomized trial is needed to clarify its prognostic value. PMID- 10461057 TI - Dose intensification of platinum compounds with glutathione protection as induction chemotherapy for advanced ovarian carcinoma. AB - Based on previous clinical experience indicating the tolerability and efficacy of high-dose cisplatin with glutathione protection in the treatment of advanced ovarian cancer, this study was undertaken to explore the efficacy and feasibility of an alternative high-dose, platinum-based approach including a combination of high-dose cisplatin plus carboplatin as induction chemotherapy of advanced ovarian carcinoma and intervention surgery. Fifty consecutive eligible patients with untreated stage III or IV epithelial ovarian cancer received 40 mg/m(2) cisplatin daily on days 1-4 and 160 mg/m(2) carboplatin on day 5. The cycle was repeated after 28 days. Patients received glutathione (2,500 mg) before each cisplatin or carboplatin administration and standard intravenous hydration. After 2 courses of induction chemotherapy, the patients underwent surgical reevaluation with debulking, when possible, followed by a further 3 cycles of 120 mg/m(2) cisplatin (i.e. 40 mg/m(2) daily for 3 consecutive days plus 600 mg/m(2) cyclophosphamide on day 3) except in instances of lack of response. All eligible patients were assessed for response and toxicity. The toxicity was moderate with lack of significant nephrotoxicity. Neurotoxicity and ototoxicity were acceptable and in no patient was treatment discontinued for those toxic effects. Myelotoxicity was somewhat more severe than that observed with our previous study with high-dose cisplatin and probably related to the addition of carboplatin. Of the 40 responsive patients, 23 (46%) had a pathological complete response and 4 (8%) had a clinical complete response (without second-look laparotomy). The efficacy of the present protocol was also documented by overall survival (median survival >48 months), which appeared to be better than expected with the current therapy in this group with advanced/bulky disease. The impressive efficacy suggests a possible contribution of reduced glutathione itself in improving the outcome, as supported by preclinical studies. The results of this study should be placed in context with current platinum-based therapy including paclitaxel. PMID- 10461058 TI - Malignant seeding of the lumpectomy cavity upon breast-conserving surgery. AB - One of the etiologic factors involved in local recurrence after breast-conserving surgery may be malignant seeding of the wound during the lumpectomy procedure. A total of 340 patients with stage I and II breast cancer were entered into the study. Of these, 270 patients received breast-conserving surgery (BCS group), and the other 70 patients underwent mastectomy (control group). After resection, lavage cytology was performed at the surgical wound. There were 55 patients (20.4%) who showed positive lavage cytology in the BCS group. In the control group, there were only 3 patients (4.3%) with positive cytology. Positivity was significantly higher in the former group (p = 0.00064). Patients with evidence of cutting across cancer lesions showed significantly higher positive rates in lavage cytology (p < 0.00001). Positivity in lavage cytology was significantly higher in patients with positive surgical margins evaluated by frozen sections (p = 0.0017), touch cytology (p < 0.0001) and formalin-fixed, paraffin-embedded sections (lateral or medial margin; p = 0.0036, anterior and posterior margin: p = 0.0210). The positivity was also significantly higher in patients with an extensive intraductal component (p < 0.0001), and less than or equal to 50 (p = 0.0061) years of age. Multivariate analysis revealed that the highest relative risk factor for positive cytology was evidence of cutting across cancer lesions (relative risk = 8. 166; p < 0.00001). PMID- 10461059 TI - Outpatient treatment of neutropenic fever with oral antibiotics and granulocyte colony-stimulating factor. AB - In recent years, several cancer patients who developed neutropenic fever were effectively treated on an outpatient basis with either intravenous or oral antibiotics. This approach is associated with reduced cost and improved patient convenience. However, the appropriate antibiotic regimen and the role of growth factors have not been established yet. In order to address these issues we performed a nonrandomized phase II study to assess the feasibility and efficacy of an oral antibiotic regimen in combination with granulocyte colony-stimulating factor (G-CSF) for the outpatient treatment of cancer patients with low-risk neutropenic fever. In 50 patients with solid tumors or lymphoma, 60 episodes of neutropenic fever were treated with the combination of oral ofloxacin 400 mg twice a day, oral amoxicillin 1 g 3 times a day and G-CSF 5 microgram/kg/day subcutaneously. Patients receiving G-CSF prophylaxis were eligible for our study. Oral antibiotics were administered for at least 5 days and G-CSF was continued until resolution of neutropenia. Our patients were ambulatory, hemodynamically stable, and without significant comorbidity. Our combination was successful in 57 episodes (95%) with a median time for fever resolution of 3 days (range: 1-5 days). There was no significant toxicity associated with the antibiotic regimen with the exception of one case of reversible renal impairment. The role of G-CSF in the success of our antibiotic treatment is highly questionable since one half of our patients developed fever while on G-CSF prophylaxis. The combination of oral ofloxacin and amoxicillin with G-CSF is highly effective for the outpatient treatment of cancer patients who develop uncomplicated febrile neutropenia. The relative contribution of G-CSF needs clarification with a prospective randomized study. PMID- 10461060 TI - Cooccurrence of reduced expression of alpha-catenin and overexpression of p53 is a predictor of lymph node metastasis in early gastric cancer. AB - BACKGROUND AND OBJECTIVES: Even though the pathological background contributes to lymph node metastasis, the biological characteristics of tumors have also gained wide attention. In this study, the expression of the cadherin-catenin complex and p53 was studied in early gastric cancer. Their correlation with lymph node metastasis and the predictability of lymph node metastases, by combining these factors, were also discussed. METHODS: One hundred and one specimens obtained from surgery were studied by immunohistochemistry using monoclonal anti-E cadherin, anti-alpha-catenin and anti-p53 antibodies. RESULTS: Expression of E cadherin and alpha-catenin was reduced in 50.5 and 64.4%, respectively. p53 protein staining was positive in 29.7%. There was a significant correlation between E-cadherin and alpha-catenin expression, but no correlation was found between p53 expression and E-cadherin or alpha-catenin expression. A reduction in alpha-catenin expression and p53 overexpression correlated to lymph node metastases, respectively. Multivariate analysis showed that cooccurrence of reduced expression of alpha-catenin and overexpression of p53 was an independent factor indicating lymph node metastases. CONCLUSION: A study of both alpha catenin and p53 expression may be helpful to predict lymph node metastases in early gastric cancer. PMID- 10461061 TI - Tumor-associated macrophage infiltration in pulmonary adenocarcinoma: association with angiogenesis and poor prognosis. AB - The relation between tumor-associated macrophage (TAM) density and the density of microvessels was investigated in specimens from 113 patients with pulmonary adenocarcinoma, as was the influence of TAM density on prognosis. The rank correlation test revealed a significant relation between TAM density and microvessel density (y = 14.418 + 0.863x, r = 0.454, p > 0.0001). A significant difference in patient survival rate was detected between tumors with a TAM density defined as high and those with a TAM density defined as low (p < 0.01). Multivariate analysis also showed that TAM density was significantly related to survival (p < 0.05). These data indicate that TAM infiltration may contribute to tumor angiogenesis, and that TAM density is a useful prognostic marker in pulmonary adenocarcinoma. PMID- 10461062 TI - Serum pro-gastrin-releasing peptide is a useful marker for treatment monitoring and survival in small-cell lung cancer. AB - We investigated the usefulness of serum pro-gastrin-releasing peptide (Pro-GRP) as a tumor marker for diagnosis, treatment monitoring and the prediction of relapse and prognosis in patients with small-cell lung cancer (SCLC). Serum samples were obtained from 127 patients with primary lung cancer (48 patients with small-cell carcinoma, 31 with adenocarcinoma, 36 with squamous cell carcinoma and 11 with large-cell carcinoma). The cutoff levels of serum Pro-GRP and neuron-specific enolase (NSE) were set at 46 pg/ml and 10 ng/ml, respectively. The specificity of Pro-GRP was significantly higher than that of NSE (Pro-GRP: 93.7%, NSE: 65.8%, p < 0.01). According to the histological type of lung cancer, the positive rates of Pro-GRP were 75% (36/48) in the small-cell carcinomas, 9.7% (3/31) in the adenocarcinomas, 5.6% (2/36) in the squamous cell carcinomas and 0% (0/10) in the large cell carcinomas. The median levels of Pro GRP in limited disease (LD) and extensive disease (ED) patients were 199 and 295.5 pg/ml, whereas those of NSE were 14.8 and 29.3 ng/ml, respectively. The positive rates of Pro-GRP in LD and ED patients were 80.0% (16/20) and 71.4% (20/28), whereas those of NSE were 70.0% (14/20) and 89.3% (25/28), respectively. The positive rate of NSE tended to elevate with the progression of disease, whereas that of Pro-GRP was already high at an early stage. Among the 29 patients with SCLC who could be followed, the serum Pro-GRP levels of 18 responders were significantly decreased after treatment (p < 0.01), whereas those of the 11 nonresponders were not significantly different between before and after treatment (p = 0.72). In the 9 patients with SCLC who relapsed, the serum Pro-GRP levels were again elevated at the time of relapse. Seventeen patients whose ratio of the Pro-GRP level after treatment to the level before treatment was below 50% (taking the levels before treatment as 100%) survived significantly longer than did the patients whose ratio was over 50% (p < 0.01). The results of the present study suggest that serum Pro-GRP has high specificity and could be a useful marker of SCLC for treatment monitoring and prognosis. PMID- 10461063 TI - Hypermethylation of the p16 gene in sporadic T3N0M0 stage colorectal cancers: association with DNA replication error and shorter survival. AB - Hypermethylation in the promoter region of the p16 gene was suspected to be involved in the tumorigenesis of colorectal cancers, although its clinical and biological significance remains obscure. In this study, we collected 84 T3N0M0 stage primary colorectal cancers that were curatively resected. The clinicopathologic data were reviewed. p16 hypermethylation was determined by a methylation-specific polymerase chain reaction (PCR). p53 overexpression was detected by immunocytochemistry (ICC). The point mutations in the 12 and 13 codons of the K-ras gene were screened by restriction enzyme analysis. Loss of heterozygosity (LOH) of the DCC (Deleted in Colorectal cancer) gene was examined by PCR using primers of the DCC (18q21) microsatellite marker. The DNA replication error (RER) was examined using 7 microsatellite markers at distinct chromosomal loci. p16 hypermethylation, regarded as an indication of p16 inactivation, was evident in 24 (28.6%) of the tumors. No correlation was found between p16 hypermethylation and various clinicopathologic factors, includinig age, sex, tumor location, tumor size, growth pattern, tumor differentiation, mucin production, vascular and/or lymphatic invasion, lymphocyte infiltration of the tumor, and serum level of carcinoembryonic antigen. There was no association between p16 hypermethylation of K-ras gene mutation, p53 overexpression and LOH of the DCC gene. However, p16 hypermethylation was significantly associated with DNA RER (p = 0.01). Survival analysis revealed a significant survival disadvantage of p16-hypermethylated versus non-p16-hypermethylated tumors (p = 0.0001). These findings indicate that p16 hypermethylation plays a role in the carcinogenesis of a subset of colorectal cancers; and the presence of p16 hypermethylation predicts shorter survival in T3N0M0 stage colorectal cancers. PMID- 10461064 TI - Amplification of c-myc in hepatocellular carcinoma: correlation with clinicopathologic features, proliferative activity and p53 overexpression. AB - Expression of the proto-oncogene c-myc has been implicated in liver regeneration and hepatocarcinogenesis. The biologic significance of c-myc gene amplification in human hepatocellular carcinoma, however, is unconfirmed. We correlated c-myc gene amplification with clinicopathologic features, proliferative activity, and p53 expression in 42 resected tumors. c-myc amplification in tumor tissue was determined using a differential polymerase chain reaction, a useful procedure for the evaluation of gene amplification in archival formalin-fixed paraffin-embedded tissues, in comparison with a dopamine D2 receptor gene. Proliferative activity was estimated by numbers of argyrophilic nucleolar organizer regions and immunohistochemical nuclear labeling rates using a monoclonal antibody against Ki 67. The c-myc gene was amplified in 14 of 42 tumors (33.3%). Amplification of c myc was more frequent in younger patients and in larger tumors, and less differentiated tumors. No correlation was noted with alpha-fetoprotein level or viral hepatitis state. The amplification showed positive correlation with both proliferative activity and p53 overexpression. Disease-free survival in patients showing c-myc amplification was significantly shorter than in those without amplification. These results suggest that c-myc amplification is an indicator of malignant potential and poor prognosis in hepatocellular carcinoma. c-myc amplification and p53 alteration may be coparticipating events in the progression of these tumors. PMID- 10461065 TI - Neoangiogenesis and relationship to nuclear p53 accumulation and vascular endothelial growth factor expression in advanced gastric carcinoma. AB - Recent studies have shown that tumor suppressor genes, particularly p53, may play a crucial regulatory role in the control of angiogenesis. Accordingly, we analyzed the correlation between p53 accumulation and the expression of vascular endothelial growth factor (VEGF) and microvessel counts in a series of human gastric adenocarcinomas by immunohistochemical staining. p53 expression was absent in 35 cases (29.7%), weak in 34 cases (28.8%), moderate in 35 cases (29.7%), and strong in 14 cases (11.8%). The prognosis of patients with both absent and weak expression of p53 was significantly better than that of patients with both moderate and strong expression of p53. VEGF expression was significantly associated with increased p53 expression. Moreover, increasing expression of p53 significantly correlated with an increase in microvessel counts. Nuclear p53 accumulation may correlate with tumor angiogenesis through VEGF upregulation in gastric carcinomas. PMID- 10461066 TI - Low incidence of germline BRCA2 gene mutations among Spanish breast cancer patients. PMID- 10461067 TI - Role of the combination of ifosfamide and etoposide in the treatment of soft tissue sarcomas. PMID- 10461068 TI - Preclinical evaluation of benzoporphyrin derivative combined with a light emitting diode array for photodynamic therapy of brain tumors. AB - OBJECTIVE: The aim of this study was to investigate the second-generation photosensitizer benzoporphyrin derivative (BPD) and a novel light source applicator based on light-emitting diode (LED) technology for photodynamic therapy (PDT) of brain tumors. METHODS: We used a canine model to investigate normal brain stem toxicity. Twenty-one canines underwent posterior fossa craniectomies followed by PDT with BPD. These animals were compared to light only and BPD control. In addition, we investigated the ability of BPD and LED to cause inhibition of cell growth in canine glioma and human glioma cell lines, in vitro. The biodistribution of BPD labeled with 111In-BPD in mice with subcutaneous and intracerebral gliomas and canines with brain tumors was studied. RESULTS: The in vivo canine study resulted in a maximal tolerated dose of 0.75 mg/kg of BPD and 100 J/cm(2) of LED light for normal brain tissue. The in vitro study demonstrated 50% growth inhibition for canine and human glioma cell lines of 10 and 4 ng/ml, respectively. The mucine study using 111In-BPD showed a tumor to normal tissue ratio of 12:1 for intracerebral tumors and 3.3:1 for subcutaneous tumors. Nuclear scans of canines with brain tumors showed uptake into tumors to be maximal from 3 to 5 h. CONCLUSION: Our study supports that BPD and LED light sources when used at appropriate drug and light doses limit normal brain tissue toxicity at doses that can cause significant glioma cell toxicity in vitro. In addition, there is higher BPD uptake in brain tumors as compared to normal brain in a mouse glioma model. These findings make BPD a potential new-generation photosensitizer for the treatment of childhood posterior fossa tumors as well as other malignant cerebral pathology. PMID- 10461069 TI - Immediate posttraumatic seizures: is routine hospitalization necessary? AB - OBJECTIVE: A recent Internet survey of pediatric neurosurgeons showed that 86% routinely admitted children with immediate posttraumatic seizures (PTS) for a brief period of observation. We wished to determine whether certain children meeting predefined criteria could instead be safely discharged from the emergency room. METHODS: We reviewed the records of children admitted during the past 5 years with a diagnosis of seizure and head injury. Children with a minor head injury, a PTS occurring within 24 h of injury and no intracranial abnormalities on admission CT scan were included. Children with previous neurological conditions, a history of prior seizures (other than PTS or febrile seizures), a prior history of anticonvulsant use, or intracranial abnormalities on the admission CT scan were excluded. Records were abstracted for child's age, gender, length of admission, previous history of PTS or febrile seizures, mechanism of injury, location of impact, time between impact and PTS, the number, length and type of PTS, Glasgow Coma Score (GCS) on admission, subsequent complications and hospital costs. RESULTS: Seventy-one children met the inclusion criteria. Eleven children presented to the emergency room with prolonged seizures, transient apnea or persistently low GCS and required admission to the intensive care unit (ICU). Among the 60 remaining children with simple PTS, none had further seizures during the follow-up period, and none had significant complications. The average cost of hospitalization was known for 58 children; after excluding the costs for 5 patients who were admitted to the ICU, the average hospital cost amounted to USD 1,615 per patient. CONCLUSIONS: Our data suggest that children with isolated minor head injuries and simple PTS who recover fully in the emergency room, whose CT scans show no intracranial abnormalities and who have no prior history of neurological disease, epilepsy or anticonvulsant use are at low risk for recurrent seizures or neurological complications, and could potentially be sent home to a reliable caretaker and a stable home situation. However, because of the limited sample size in this study, the statistical risk of a bad outcome may be as high as 9%; we therefore suggest that much larger studies are potentially needed before this becomes a standard policy. PMID- 10461070 TI - Pediatric blunt carotid injury: a review of the National Pediatric Trauma Registry. AB - Blunt carotid injury (BCI) is an uncommon yet potentially devastating entity which has received little attention in the pediatric literature. In an attempt to better characterize pediatric BCI, a review of the National Pediatric Trauma Registry was performed. Records were obtained from all children diagnosed with internal or common carotid injury associated with blunt trauma. The incidence of BCI was 0.03% (15 of 57,659 blunt trauma patients). Variables examined included: age, gender, mechanism of injury, associated injuries, various injury severity scores, and outcome. Various injuries were associated with an increase in BCI incidence including chest trauma (4-fold), combined head and chest trauma (6 fold), basilar skull fractures (4-fold), intracranial hemorrhage (6-fold), and clavicle fractures (8-fold). Thirty-three percent of the patients diagnosed with BCI suffered neurological complications directly attributable to their carotid injuries. Current practices regarding screening, diagnosis, and treatment are reviewed. PMID- 10461071 TI - The bifrontal olfactory nerve-sparing approach to lesions of the suprasellar region in children. AB - Suprasellar masses in children include lesions such as craniopharyngiomas and germ cell neoplasms. The difficult location of these lesions and their proximity to important neural and vascular structures pose challenges to resection. We operated on 14 patients using a bifrontal craniotomy with removal of both orbital rims to provide access to suprasellar masses, even those with significant extension into the third ventricle. A complete resection was possible in 8/14 patients and 8/10 craniopharyngiomas. In 13 patients, the optic nerves were identified and preserved, and vision was stable or improved postoperatively. Postoperatively, 1 patient with hydrocephalus developed a CSF leak which was successfully treated with a ventriculoperitoneal shunt. No patient had a cosmetic defect related to orbital rim removal. The bifrontal approach used here enhanced the exposure of the suprasellar region and minimized manipulation of the optic apparatus and the carotid arteries. It may be used alone, or in conjunction with other approaches, to resect lesions in the suprasellar region. PMID- 10461072 TI - Does age or other factors influence the incidence of ventriculoperitoneal shunt infections? AB - Some studies indicate that infants, especially those less than 1 month of age have a higher incidence of ventriculoperitoneal shunt infections. To look at age as well as other variables that might relate to the rate of shunt infection, we reviewed the records of all patients undergoing a ventriculoperitoneal shunt insertion or revision at our institution from January 1, 1985, to December 31, 1994. There were a total of 2,325 ventriculoperitoneal shunting procedures performed on 1,193 patients with a male:female ratio of 678:515. The overall infection rate was 3.2% (74 infections). Analyzed by age, the infection rates were as follows: <1 month 9/223 (4.0%), 1-6 months 16/449 (3.6%), 6-12 months 13/297 (4.4%), 12-18 months 3/122 (2.5%), 18-24 months 7/116 (6.0%) and 24+ months 26/1, 118 (2.3%). There was no statistically significant difference between age groups (p > 0.05). Upon selectively examining premature neonates who developed hydrocephalus secondary to intraventricular hemorrhage from the figures given above, one finds that 2/44 (4.5%) of neonates became infected, which was also not significant. The infection rate was the same irrespective of whether the procedure was to insert or revise the shunt, or whether another operative procedure was done under the same anesthesia. The etiology of the hydrocephalus was not a factor, nor was the presence of an open neural tube defect. The presence of fluid accumulation along the shunt tract or at another neurological operative site was associated with a significant increase in incidence of infection 15/168 (8.9%) when compared to those with no fluid accumulation (p < 0.001). The type of infecting organism was divided roughly in thirds, with relatively equal representation from Staphylococcus epidermidis/coagulase negative and Staphylococcus aureus. The remaining third was comprised of a wide variety of organisms. PMID- 10461073 TI - Embolization of a giant torcular dural arteriovenous fistula in a neonate. AB - We treated a neonatal infant who presented with heart failure and a giant torcular dural arteriovenous fistula by staged transtorcular embolization with two guidewires, 95 platinum microcoils, and tissue glues through a needle-size craniotomy. Blood loss was minimal. The patient was stable without heart failure after a three-staged embolization. The lesion disappeared on the follow-up angiography done at the age of 3 years. This is the first documented case of neonatal giant torcular dural arteriovenous malformation cured by interventional neuroradiology. The dilemma and strategy in managing this patient will be presented and discussed. PMID- 10461074 TI - Tandem spinal cord injuries without radiographic abnormalities in a young child. AB - Anatomic features unique to the pediatric spine render this population susceptible to spinal cord injuries without radiographic abnormalities (SCIWORA). To date, published descriptions of SCIWORA have been limited to a single region of the spinal column. We describe a case of a 3-year-old boy in whom, after a motor vehicle accident, tandem SCIWORA lesions involving the lower cervical spinal cord and thoracolumbar junction resulted in severe quadraparesis. The child was initially treated with 24 h of methylprednisolone followed by 3 months of external orthoses of both the cervical and thoracic spine. We include in this article a brief review of the literature and treatment guidelines for SCIWORA and postulate the mechanism of these tandem spinal cord injuries. PMID- 10461075 TI - Infundibuloneurohypophysitis in children. A report of 2 cases. AB - Two children with diabetes insipidus secondary to a chronic inflammatory infiltrate of the infundibulum - infundibuloneurohypophysitis - are presented. Features unique to these cases are contrasted with prior case reports. The differential diagnoses of diabetes insipidus and inflammatory pituitary disorders are discussed. Recommendations for diagnosis and treatment are proposed. PMID- 10461076 TI - Segmental costovertebral malformations: association with neural tube defects. Report of 3 cases and review of the literature. AB - Patients with spondylocostal dysostosis (SCD) have vertebral abnormalities and numerical or structural rib anomalies that produce thoracic asymmetry. Rib anomalies and dysmorphism are the typical features that differentiate this syndrome from spondylothoracic dysostosis (STD). Jarcho-Levin syndrome is a severe form with involvement of the whole vertebral column. Other associated findings such as congenital heart defects, abdominal wall malformations, genitourinary malformations and upper limb anomalies may be found; in addition, neural tube defects (NTDs) have been associated with this malformation. SCD is transmitted both in a recessive form and as a dominant defect. We report on 3 children with SCD; 2 also had NTDs. All of them were studied with X-rays and spinal magnetic resonance (MR), and over the same period they underwent multidisciplinary clinical functional evaluation. One of our cases with NTD also presented polythelia, which has not previously been described in patients with SCD. The common association of segmental costovertebral malformations with NTDs could be related to an early gastrulation genomic defect, or one after gastrulation, when there are two independent somitic columns. The latter sometimes progresses and then involves primary and secondary neurulation. Also, the association of SCD with NTDs could be related to the interaction of different genes, resulting in this complex phenotype. Therefore, additional genetical and embryological studies are necessary to provide evidence of an etiological link between SCD and NTD. PMID- 10461077 TI - Coexistence of a ganglioglioma and Rasmussen's encephalitis. AB - A small number of recent reports have documented coexisting cerebral pathologic entities in patients with Rasmussen's encephalitis. We report the case of a 4 year-old boy who presented with refractory seizures and was found to have both a ganglioglioma and Rasmussen's encephalitis of the same hemisphere. The patient ultimately underwent a functional hemispherectomy with excellent seizure control. We also review the clinical and pathologic hallmarks of both gangliogliomas and Rasmussen's encephalitis. We hope that the addition of this patient to the small but growing literature on dual pathology in Rasmussen's encephalitis may help shed some light on the etiology of this mysterious disease. PMID- 10461078 TI - Pulmonary malignancies in the immunocompromised patient. AB - Clinicians should be familiar with immunodeficiency-related malignancies, as their incidence is expected to increase further with the rise in the number and survival of immunocompromised patients. The most common malignancies affecting the lungs in those patients are Kaposi's sarcoma, non-Hodgkin's lymphoma and, to a far less extent, Hodgkin's disease and bronchogenic carcinoma. However, their relative frequency depends on the types of immune deficiency, including those due to congenital disorders, AIDS and drug treatments. This review will summarize epidemiological data on the frequency of immmunodeficiency-related malignancies, recent advances on their pathogenesis and current approaches to their diagnosis and treatment in the various immunosuppressed groups. PMID- 10461079 TI - Prevalence of sleep disordered breathing and sleep apnea in 50- to 70-year-old individuals. A survey. AB - PURPOSE: To determine the prevalence of sleep disordered breathing (SDB) and sleep apnea syndrome (SAS) in a general population aged from 50 to 70 years. SUBJECTS AND METHODS: We recruited 76 individuals aged between 50 and 70 years, chosen at random from the electoral census. They were invited to the clinic where a detailed medical history was taken and physical examination, ENT examination, pulmonary function tests and night time recording of respiratory variables performed. RESULTS: The prevalence of SDB (apnea-hypopnea index >/= 5) was 28.9%, and there were no differences between men (28%) and women (30%). However, the prevalence of SAS was 6.8%, and there were differences between men (5 cases) and women (0 cases) (p = 0.0521). Subjects in the SDB group had higher systolic blood pressure than in the non-SDB group (p < 0.05). CONCLUSIONS: SDB and SAS are common among 50- to 70-year olds. The prevalence of SDB was 28.9% and the prevalence of SAS was 6.8%. PMID- 10461080 TI - Reliability of thoracic gas volume derived from mechanical impedance at different levels of the vital capacity. AB - BACKGROUND: Thoracic gas volume (TGV) may be estimated during spontaneous breathing by measuring simultaneously respiratory impedance (Zrs) and alveolar gas compression (Vpl) at several oscillation frequencies [Peslin and Duvivier: J Appl Physiol 1998;84:862-867]. OBJECTIVE: The aim of the study was to test the validity of that approach at different levels of the vital capacity (VC). METHODS: We measured Zrs and Vpl at frequencies ranging from 6 to 29 Hz in 10 healthy subjects rebreathing BTPS gas in a constant volume body plethysmograph. In a first series, the subjects were asked to breathe voluntarily at different levels of the VC and oscillation TGV (TGVos) was compared to standard plethysmographic TGV (TGVst) assessed immediately after TGVos measurements. In a second series, the subjects were asked to change stepwise their lung volume in the middle of the forced oscillation recording, and the changes in TGVos (DeltaTGVos) were compared to the changes in lung volume (DeltaV) computed from the integrated flow signal. RESULTS: In most subjects TGVos and TGVst were highly correlated and the slopes of the relationships did not differ significantly from unity. DeltaTGVos and DeltaV were also highly correlated both in individuals and in the group (r = 0.97), and their signless differences averaged 0.23 +/- 0.20 liter. CONCLUSION: We conclude that forced oscillation estimates of TGV are reliable in healthy subjects over a large part of the VC. PMID- 10461081 TI - Inhaled and systemic corticosteroid therapies: Do they contribute to inspiratory muscle weakness in asthma? AB - BACKGROUND: Patients with asthma incur the risk of steroid-induced myopathy, which is a well-known side effect of treatment with corticosteroids. However, the adverse effect of long-term steroid treatment on respiratory muscle function remains controversial. OBJECTIVE: We aimed to evaluate the effects of long-term moderate dose of systemic corticosteroids and high-dose inhaled beclomethasone on maximal inspiratory and expiratory pressures (PImax and PEmax, respectively) in two groups of asthmatic patients exhibiting comparable levels of hyperinflation. METHODS: Twelve steroid-dependent asthmatic patients requiring 10-20 mg/day of prednisone-equivalent corticosteroids for an average of 9.83 +/- (SD) 9.86 years; 14 subjects with moderate to severe asthma who have used inhaled beclomethasone for at least 1 year at a daily dose higher than 1,000 microg and 15 healthy controls were included to the study. RESULTS: No significant difference in pulmonary function tests and arterial blood gases appeared between two asthmatic groups with different treatment modalities. PImax as an absolute value was significantly lower in steroid-dependent asthmatics than in patients treated with inhaled beclomethasone and controls (p < 0.01). %PImax was also lower in steroid dependent asthmatics than in control groups (p < 0.01). A significant correlation was found between %PImax and hyperinflation assessed by %RV, %FRC, %FRC/TLC (p < 0.05) in all asthmatic patients. CONCLUSIONS: We believe that hyperinflation plays a major role in inspiratory muscle dysfunction in asthma, but the finding of significantly decreased PImax values in steroid-dependent asthmatics when compared with patients on high-dose inhaled beclomethasone with a comparable level of hyperinflation points to a deleterious effect of long-term, moderate dose systemic corticosteroid but not high-dose beclomethasone on inspiratory muscle function in asthmatics. PMID- 10461082 TI - Pneumoconiosis among Cretan dental technicians. AB - Pneumoconiosis among dental technicians has recently emerged as an area of research in interstitial lung disease. This study was carried out to estimate the prevalence of pneumoconiosis in Greek dental technicians on the island of Crete. Fifty-one of the 58 dental technicians of the Heraklion Dental Technicians' Association completed an exposure history questionnaire and underwent a clinical examination, including chest radiographs, and spirometric assessment of lung volume and diffusing capacity. Values were compared with 51 control subjects. Five dental technicians showed radiological evidence of pneumoconiosis. Mean lung function values of the dental technicians, even in those with pneumoconiosis, were not significantly different from controls. No significant changes in lung function parameters were associated with smoking, exposure to metals and other contaminants. Dental technicians, however, reported more respiratory symptoms than controls (p = 0.008). Symptoms were associated to the absence of a ventilation system. We concluded that occupational exposure in dental technicians in Crete did not affect lung function. The prevalence of pneumoconiosis in this study was 9. 8%. PMID- 10461083 TI - Immunohistochemical localization of B7 costimulating molecules and major histocompatibility complex class II antigen in pulmonary sarcoidosis. AB - BACKGROUND: Alveolar macrophages (AM) of sarcoidosis have an enhanced capacity to mediate antigen-induced T lymphocyte proliferation. To induce an effective immune response, antigen-presenting cells have to not only present antigenic peptide with MHC molecules to T lymphocytes, but also express B7 costimulating molecules. OBJECTIVE: The purpose of this study was to investigate the expression of B7 and MHC molecules in lung tissues from patients with sarcoidosis. METHODS: We performed immunohistochemistry for B7-1, B7-2 and MHC class II antigens using transbronchial lung biopsy specimens obtained from patients with sarcoidosis and normal lung parenchyma obtained by lobectomy for solitary pulmonary nodule as controls. RESULTS: B7-1, B7-2 and MHC class II antigen were expressed in epithelioid cells in granulomas in 14 (93.3%), 2 (13.3%) and 9 (60.0%) of 15 patients with sarcoidosis, respectively. These were also expressed in AM in 14 (93. 3%), 5 (33.3%) and 12 (80.0%) of 15 patients with sarcoidosis, respectively. The positivity of B7-1 was significantly higher than that of B7-2 in both epithelioid cells and AM in sarcoidosis (p < 0. 01). Positive signals for B7-1, B7-2 and MHC class II antigen were also found in AM in 9 (90%), 8 (80%) and 8 (80%) of 15 of controls, respectively. However, the intensity of positive signals for B7-1, but not B7-2 or MHC class II antigen in AM was significantly increased in sarcoidosis compared to controls (p < 0.05). CONCLUSIONS: These results suggested that epithelioid cells in granulomas and AM from patients with sarcoidosis had the capability to act as accessory cells and that the accessory function of these cells was shifted to B7-1 rather than B7-2 in sarcoidosis. PMID- 10461084 TI - Both inhaled histamine and hypertonic saline increase airway reactivity in non sensitised rabbits. AB - BACKGROUND: Asthmatics react with bronchoconstriction upon a variety of stimuli, i.e. exercise and hypertonic aerosol challenge. We have previously shown that hyperventilation with dry gas in a rabbit model resulted in a change of the ion content of the tracheal wall. This was followed by a hyperreactive response to histamine. OBJECTIVE: We hypothesised that nebulisation with 3.6% hypertonic saline will be accompanied by a hyperreactive response to histamine in a rabbit model. METHODS: Anaesthetised rabbits were given histamine after nebulisation with hypertonic saline. In addition, repeat nebulisation with hypertonic saline was given with or without histamine between these nebulisations. RESULTS: There was a different response to histamine 10 mg x ml(-1) whether hypertonic saline had been given or not (p < 0.001). Histamine nebulisation, given after hypertonic saline, caused an increase from baseline in resistance of 65 +/- 12 cm H(2)O.litre(-1) x s (mean +/- SEM, p < 0. 001) and a decrease in compliance of 2.3 +/- 0.4 ml x cm H(2)O(-1) (p < 0.001). The corresponding values for the control animals were 10 +/- 4 cm H(2)O.litre(-1) x s (n.s.) and 1.7 +/- 0.2 ml x cm H(2)O(-1) (p < 0.001). At a second nebulisation with hypertonic saline, with a histamine challenge 30 min before, the resistance increased from baseline by 35 +/- 10 cm H(2)O x litre(-1) x s (p < 0.01). This was not observed when no histamine had been given between the hypertonic saline nebulisations. CONCLUSIONS: This study in rabbits shows that hypertonic solutions cause an increase in the responsiveness to histamine and that histamine causes an increase in responsiveness to hypertonic saline. This is similar to the response of asthmatics to hypertonic saline. PMID- 10461085 TI - Substance P relaxes rat bronchial smooth muscle via epithelial prostanoid synthesis. AB - BACKGROUND: Substance P is present in bronchial nerve fibres. The physiological actions of substance P are mediated via tachykinin NK(1) receptors. Immunochemical studies have demonstrated tachykinin NK(1) receptors in the rat airway epithelium. OBJECTIVE: To elucidate how epithelial tachykinin NK(1) receptors affect smooth muscle response to substance P. METHODS: Contractile response of isolated rat bronchial trunk with or without epithelium was recorded. RESULTS: In intact segments precontracted by 5-hydroxytryptamine, relaxation was induced by substance P and the nitric oxide donor, sodium nitroprusside. Removal of the epithelium abolished relaxation induced by substance P but did not affect relaxation induced by sodium nitroprusside. The cyclo-oxygenase inhibitor, indomethacin, but not the nitric oxide synthase inhibitor, L-N(G) monomethylarginine, reduced the relaxation in response to substance P. CONCLUSIONS: Epithelial tachykinin NK(1) receptors mediate substance-P-induced relaxation of rat bronchial smooth muscle via release of prostanoids but not nitric oxide. PMID- 10461086 TI - The eye catcher. Reversible lungs or reversed X-ray? PMID- 10461087 TI - Congenital broncho-oesophageal fistula associated with bronchiectasis in adults. Report of two cases and review of the literature. AB - Congenital broncho-oesophageal fistula is a rare entity in adult patients. This anomaly may cause various symptoms such as respiratory infections, coughing bouts when eating or drinking and even haemoptysis. Even rarer than its occurrence with the above-mentioned symptoms is its presentation with bronchiectasis. A congenital broncho-oesophageal fistula presenting with bronchiectasis in a 28 year-old male and 36-year-old female are described. In reported cases, symptoms of chronic recurrent pulmonary suppuration were initially attributed to alternative aetiologies. In both cases, with such an unusual presentation, the observation of the fistulous tract was coincidental. Surgical division of the fistula associated with lobectomy resulted in complete resolution of symptoms. PMID- 10461088 TI - Pleural empyema: An unusual presentation of esophageal perforation. AB - A 67-year-old patient presented with pleural empyema as the sole manifestation of thoracic esophageal perforation, 2 weeks after accidental fish bone ingestion. Nonspecific chest pain and general deterioration, unusual presenting symptoms in themselves, accounted for the extreme delay in the diagnosis. The empyema was treated surgically, and the esophageal perforation conservatively. Despite the poor prognostic factors, the patient recovered completely after 50 days in hospital. PMID- 10461089 TI - Allergic bronchopulmonary aspergillosis due to Aspergillus niger without bronchial asthma. AB - A 65-year-old woman was admitted to our hospital with a dry cough and pulmonary infiltrates. Chest radiograph and CT revealed mucoid impaction and consolidations. Peripheral blood eosinophilia and elevated serum IgE were observed. Aspergillus niger was cultured repeatedly from her sputum, but A. fumigatus was not detected. Immediate skin test and specific IgE (RAST) to Aspergillus antigen were positive. Precipitating antibodies were confirmed against A. niger antigen, but not against A. fumigatus antigen. She had no asthmatic symptoms, and showed no bronchial hyperreactivity to methacholine. Thus, this case was diagnosed as allergic bronchopulmonary aspergillosis (ABPA) without bronchial asthma due to A. niger, an organism rarely found in ABPA. The administration of prednisone improved the symptoms and corrected the abnormal laboratory findings. PMID- 10461090 TI - Massive pulmonary hemorrhage due to cytomegalovirus infection in a Japanese patient with alpha-1-antitrypsin-deficient emphysema. AB - Although alpha(1)-antitrypsin (AAT) deficiency is one of the most common hereditary diseases and a recognized cause of emphysema in Caucasians, variants of this deficiency are extremely rare among Orientals. We present here a Japanese emphysema patient with the AAT deficiency variant originally identified as S(iiyama). After an 8-year follow-up period, the patient suffered from repeated pulmonary Pseudomonas aeruginosa infection for 4 years. He died suddenly of massive pulmonary hemorrhage. The pathologic examination revealed a necrotic hematoma in the right S10 lobe, which exhibited pneumonia due to cytomegalovirus (CMV) infection. Pulmonary hemorrhage due to CMV can occur and be fatal in patients with emphysema and AAT deficiency. PMID- 10461091 TI - A 59-year-old female with increasing dyspnoea, an unusual rash and myalgia. Diagnosis: dermatomyositis with associated interstitial lung disease. PMID- 10461092 TI - The effect of liposomes on skin barrier structure. AB - The present work deals with the 'in vivo' stripping technique to evaluate the percutaneous absorption of sodium fluorescein (NaFl) vehiculized in two different liposome preparations formed by phosphatidylcholine (PC) and lipids mimicking the stratum corneum (SC; ceramides, cholesterol, palmitic acid and cholesteryl sulphate), respectively. Furthermore, the possible effect of these vesicles on the SC lipid alkyl chain conformational order were evaluated at different depths of SC by non-invasive biophysical techniques: Corneometer, Tewameter and especially ATR-FTIR. The results of NaFl percutaneous absorption indicate the highest penetration in the case of incorporation in PC liposomes, which could be related to the increase in SC lipid disorder detected by ATR-FTIR, i.e. a decrease in skin barrier function. On the other hand, SC lipid liposomes have been shown to have a higher affinity for SC owing to the high amount of NaFl found in this layer, suggesting a greater reservoir capacity of SC when similar lipid composition formulation is applied. A lipid order increase is observed by infrared spectroscopy, when these types of liposomes are topically applied, resulting in a strong barrier effect. These results could be useful in designing specific liposomal topical applications. PMID- 10461093 TI - Penetration of titanium dioxide microparticles in a sunscreen formulation into the horny layer and the follicular orifice. AB - Coated titanium dioxide (TiO2) microparticles are commonly used as UV filter substances in commercial sunscreen products. The penetration of these microparticles into the horny layer and the orifice of the hair follicle was investigated. The distribution of the microparticles in the horny layer was analyzed using the method of tape stripping in combination with spectroscopic measurements. Deeper layers of the stratum corneum were devoid of TiO(2) even after repetitive application of sunscreen preparation when analyzing interfollicular areas. Only in the areas of the pilosebaceous orifices could microparticles be identified. The penetration of TiO(2) was investigated in histological skin sections. A biopsy was taken from a skin area from which the horny layer had been removed by tape stripping. In isolated areas, a penetration of coated TiO2 into the open part of the follicle was observed. The amount of TiO2 found in a given follicle was less than 1% of the applied total amount of sunscreens. A penetration of microparticles into viable skin tissue could not be detected. PMID- 10461095 TI - Long-term stability of 8-methoxypsoralen in ointments for topical PUVA therapy ('Cream-PUVA'). AB - 8-Methoxypsoralen (8-MOP) is an established photochemotherapeutic agent for PUVA therapy. Recently, a so-called 'cream-PUVA' modality was introduced into therapy of psoriasis and other dermatoses. Little is known, however, about the stability of 8-MOP in ointments used for the topical application of this compound. Therefore, we investigated the long-term stability of 8-MOP in three different ointments, Unguentum Cordes(TM), Cold Cream Naturel(TM) and a water-containing gel on the basis of Carbopol 940. All three ointments were prepared with 8-MOP concentrations of 0.05 and 0.005%, and stored over 12 weeks at room temperature (19-20 degrees C) and at 5 degrees C. 8-MOP concentrations were measured at days 1, 8, 15, 29, 57 and 88 after preparation by thin-layer chromatography (TLC). The ointments were dissolved in an organic solvent, 10 microl were transferred onto the TLC plate and the chromatograms were developed first in toluene and then in toluene/ethyl acetate 2:1 v/v to resolve 8-MOP from the ointment constituents. The peak heights of 8-MOP were used for quantitation. The intraday variabilities are <3% for Unguentum Cordes and Cold Cream Naturel and <6% for the Carbopol 940 gel. The interday variabilities were <6.3% in all cases. In Unguentum Cordes and Cold Cream Naturel the concentrations of 8-MOP remain stable, but in Unguentum Cordes the emulsion began to break up after 8 weeks. In the Carbopol gel, only about 40% of the nominal concentrations of 8-MOP were found and they decrease significantly at storage at 5 degrees C. We conclude that the Carbopol gel seems to be unsuitable for PUVA therapy, whereas Cold Cream Naturel shows the best results. PMID- 10461094 TI - Influence of ion channel blockers on proliferation and free intracellular Ca2+ concentration of human keratinocytes. AB - The proliferation of in vitro cultivated keratinocytes from the HaCaT cell line while under the influence of different ion channel blockers, i.e. calcium channel blockers tetraethylammonium (TEA) and the nonspecific anion channel blocker 4',4' diisothiocyanato-stilbene-2-2' disulfonic acid (DIDS) was investigated. There were changes in proliferation of the keratinocytes brought on by TEA and DIDS. It can be assumed that the effects of TEA on proliferated keratinocytes lead to a temporary arrest of cells in the G(1)-phase of the cell cycle. The change in the extracellular K+ concentration ([K+]e) had no influence on the proliferation of keratinocytes. During the entire course of investigations, the adding of DIDS leads to an increase in proliferation. Proliferation of keratinocytes is known to be greatly dependent on intracellular Ca2+ content ([Ca2+]i). The blocking of ion channels with TEA or DIDS does not lead to a change in the free [Ca2+]i. Also a change in the [K+]e does not lead to a displacement of the [Ca2+]i of keratinocytes. PMID- 10461096 TI - The response of distant uninvolved psoriatic skin to standardised injury is not different from that in normal skin. AB - The epidermis of uninvolved psoriatic skin is characterised by a slight hyperproliferation and an increase in inflammatory parameters, whereas no differentiation abnormalities are seen. Data with respect to the response of distant uninvolved psoriatic skin to standardised injury are not uniform. In this study, a recently developed multiparameter flow cytometric assay was used to compare the response to tape stripping of uninvolved psoriatic and normal skin. With this method, a parameter for proliferation, differentiation and inflammation was measured simultaneously. Concerning these parameters, no statistically significant differences were found between uninvolved psoriatic skin and normal skin. The mechanism that underlies hyperproliferation in distant uninvolved psoriatic skin does not indicate an intrinsic abnormality in keratinocytes. Inflammatory signals might play a role in this process. PMID- 10461097 TI - N-Methyl-L-serine stimulates hyaluronan production in human skin fibroblasts. AB - We examined the effects of N-methyl-L-serine (NMS), an amino acid derivative, on hyaluronan (HA) synthesis in human skin fibroblasts. NMS (1-10 mM), but not L serine, stimulated the incorporation of [(3)H]glucosamine into HA dose dependently, with a maximum stimulation of 1.5-fold compared to the control. The effect of NMS was specific for HA production, because there was no change in sulfated glycosaminoglycan formation. Neither the N-methyl derivatives of L glycine or L-alanine, nor N-methyl-D-serine, could stimulate HA synthesis, indicating that the beta-hydroxyl group and the L-configuration were essential for the activity. Gel filtration of the products showed that NMS stimulated the production of high-molecular-mass HA (>10(6) D) without affecting the production of low-molecular-mass HA. NMS required 24 h to stimulate HA production, and when fibroblasts were pretreated for 10-24 h with NMS (1-10 mM), membrane-associated HA synthase activity was increased dose-dependently. Thus, a second messenger is likely to be involved in the stimulation of HA production by NMS. PMID- 10461098 TI - Release and skin distribution of silicone-related compound(s) from a silicone gel sheet in vitro. AB - The efficacy of topical silicone gel sheeting in prevention and/or reduction of keloids and hypertrophic scars is well recognized. In the present study, we reexamined the possible release of silicone-related compound(s) from a commercially available silicone gel sheet (Cica-Care, Smith and Nephew, Hull, England) in aqueous media in vitro. The silicone gel sheet was also applied on the excised skin surface to examine the possible distribution of silicone-related compounds into the skin in vitro. Silicone-related compounds were measured as silicon by an inductively coupled plasma-atomic emission spectrophotometer. When a piece of silicone gel sheet was placed in phosphate buffer solution (pH 3-9) at 37 degrees C for 7 days, the concentration of silicon in the medium increased with time, depending on the pH of the medium. This indicates that the released silicone-related compounds are water-soluble. When Cica-Care was applied on the surface of excised rat skin, human axilla skin and hypertrophic scars under hydrated conditions in vitro, silicon was detected in all skin samples. Greater distribution was observed in rat skin than in human axilla skin and hypertrophic scars. The release of silicone-related compounds from a silicone gel sheet (Cica Care) and their distribution into the skin were demonstrated in vitro. Silicone related compounds distributed into the skin may have pharmacological effects on the skin. Further investigation will be necessary to investigate in detail the action of silicone-related compounds on the proliferation of fibroblasts and excessive production of collagen. PMID- 10461099 TI - The effect of the base on the kinetics of action of the capsaicinoid nonivamide: evaluation with a hyperemic test. AB - Because of their selective receptor-mediated action on cutaneous nociceptive C fibers, the interest in capsaicinoids as topical analgetic drugs has been constantly growing over the past years. Knowledge of the pharmacokinetics seems particularly important for potential future clinical applications. Therefore, the aim of the present study was to investigate the influence of the type of preparation on the time course of action of the capsaicinoid nonivamide monitored by the axon-reflex-induced hyperemic action and area of erythema. Sixteen healthy subjects were included in the study. The hyperemic responses after application of nonivamide in an oil-in-water (O/W) emulsion and in a water-free ointment were assessed both by laser Doppler perfusion imaging and planimetry after 0, 15, 30, 45, 60, 120 and 240 min. They were compared with the reaction after application of the nicotinic ester nicoboxil and a combination of nonivamide and nicoboxil in the same preparations. Applied as a water-free ointment, nonivamide showed a slow onset of hyperemic action, reaching its maximum 45 min after application. When applied as an O/W emulsion, however, the increase in effect was high, reaching its maximum already after 30 min. Application of the nicoboxil preparations revealed a clearly lesser influence of the base regarding the onset of maximum effect. The combination of both substances showed an additive effect for both bases, and a maximum effect was found already after 15 min with both the water free ointment and the O/W emulsion base. In conclusion, the results of the present study show that there appears to be a strong influence of the vehicle on the kinetics of action of capsaicinoids, and that the hyperemic test used in this paper is very useful for the quantitative determination of the pharmacokinetic properties of capsaicinoids. Moreover, the C-fiber-stimulating effect of capsaicinoids can, at least in part, be enhanced by combination with a nicotinic ester, even though these substances have no direct effect on the C-fibers. PMID- 10461100 TI - Hypericin levels in human serum and interstitial skin blister fluid after oral single-dose and steady-state administration of Hypericum perforatum extract (St. John's wort). AB - The photodynamically active plant pigment hypericin, a characteristic metabolite of Hypericum perforatum (St. John's wort), is widely used as an antidepressant. When administered orally, phototoxic symptoms may limit the therapeutic use of hypericin-containing drugs. Here we describe the high-performance liquid chromatographic (HPLC) detection of hypericin and semiquantitative detection of pseudohypericin in human serum and skin blister fluid after oral single-dose (1 x 6 tablets) or steady-state (3 x 1 tablet/day, for 7 days) administration of the Hypericum extract LI 160 in healthy volunteers (n = 12). Serum levels of hypericin and pseudohypericin were always significantly higher than skin levels (p 100 ng/ml). PMID- 10461101 TI - Microelectrode recordings define the ventral posteromedial pallidotomy target. AB - The benefits of stereotactic ventral posteromedial pallidotomy in the treatment of Parkinson's disease have been recently rediscovered. Optimal lesion location and lesion volume, however, have yet to be determined. Micro-electrode recording and microstimulation are carried out to determine an appropriate site for the placement of electrocoagulation lesions in the medial pallidum. The cellular activity of the globus pallidus is examined for characteristic firing patterns, mean firing rates, movement-evoked activity, and presence of tremor cells, laminae, and border zones. Microstimulation allows the identification of the adjacent optic tract by reports of visual sensation and of the internal capsule by sensorimotor responses. Lesions are centred at sites in the internal segment of the globus pallidus at least 3 mm from these structures, to avoid injury to them during pallidotomy. PMID- 10461102 TI - Lesion volume and clinical outcome in stereotactic pallidotomy and thalamotomy. AB - Postoperative lesion volume and clinical outcome were assessed in 19 Parkinson's disease (PD) patients who received posteroventral pallidotomy, and in 14 essential tremor (ET) patients who received ventrolateral thalamotomy. Before and after surgery, PD patients were evaluated using the Unified PD Rating Scale (UPDRS), and ET patients were evaluated using the Fahn-Tolosa-Marin (FTM) tremor rating scale. Inner and total lesion volumes were determined with postoperative MR imaging and three-dimensional data segmentation. Lesion volumes were compared to percent improvement in UPDRS and FTM scores, using Spearman's rank-order correlation test. No rank-order correlations were found between lesion volume and clinical improvement in either the PD or the ET patients. In performing stereotactic surgery for movement disorders, any lesion volume within a prescribed range may be equally effective in relieving symptoms associated with PD or ET. PMID- 10461103 TI - Brain targets for pain control. AB - A variety of brain sites have been targeted for surgical treatment of intractable pain. Both ablative and chronic stimulation procedures have been reported to attenuate such pain. These targets include the thalamus and its projections, the periventricular gray, the cingulate cortex and the motor cortex. An overview of these procedures and their efficacy is provided. PMID- 10461104 TI - Direct spinal cord electrical stimulations during surgery of intramedullary tumoral and vascular lesions. AB - Despite the use of somatosensory evoked potentials during surgery for spinal cord tumors or vascular lesions, postoperative neurological disorders, particularly motor deficits, frequently occur after aggressive surgery with an attempt of gross total resection. We report the use of peroperative direct spinal cord electrical stimulation to decrease morbidity while improving the quality of resection. Three patients with intramedullary lesions (1 ependymoma and 2 cavernomas), initially revealed by pain and followed by neurological deficit, were operated at our institution using peroperative direct medullary electrical stimulations (60 Hz, biphasic square wave pulses with 1 ms/phase, 0.9 mA) under general anesthesia without curare. In all cases, gross total resection was performed until motor responses to stimulation, which indicated anterior and lateral boundaries between the lesion and the functional tissue, were obtained. There was no postoperative neurological worsening, but an immediate partial improvement of sensory and bladder disorders in the patient with ependymoma. Postoperative MRI confirmed total resection in the 3 patients. These cases demonstrate that direct medullary electrical stimulation is a safe, easy, precise and reliable method to reduce morbidity during spinal cord surgery. PMID- 10461105 TI - Effects of a stereotactic headframe assembly on proton magnetic resonance spectroscopy. AB - The effects of a magnetic resonance-compatible stereotactic headframe assembly on single voxel proton magnetic resonance spectroscopy (MRS) were investigated. Multiple stimulated echo acquisition mode (STEAM) spectra were obtained within a commercially available brain metabolite phantom placed within the headframe assembly (Leksell, Model G). All acquisition parameters were kept constant, except voxel location. Maximal distortion occurred for voxels acquired in the immediate vicinity of a headframe fixation pin, manifested by spectral broadening, changes in peak area and height and distortion of the baseline, rendering these acquisitions nondiagnostic. The range of this interaction was short, and a voxel acquired with nearest edge located 2.0 cm or greater from the fixation pin tip produced NAA/Cr, Cho/Cr and mI/Cr ratios differing by less than 7.5% from a spectrum obtained at the phantom center. The feasibility of performing single voxel MRS with a stereotactic headframe in place is demonstrated. PMID- 10461106 TI - Patterns of metastasis in muscle-invasive bladder cancer (pT2-4): An autopsy study on 367 patients. AB - BACKGROUND: Metastatic status is an essential determinant of prognosis of patients with muscle-invasive bladder cancer treated by cystectomy, and preoperative metastases detection is crucial for treatment selection in these patients. To better understand the metastatic behavior of bladder tumors, autopsies of patients with muscle-invasive bladder carcinomas (pT2-4) were evaluated. METHODS: Protocols and histologic sections from autopsies of 367 patients with pT2-4 bladder cancer were reviewed. RESULTS: Metastases were found in 251 of 367 patients (68%). The most frequent sites of metastases were regional lymph nodes (90%), liver (47%), lung (45%), bone (32%), peritoneum (19%), pleura (16%), kidney (14%), adrenal gland (14%), and the intestine (13%). There was no difference in the frequency and location of metastases between 308 transitional cell carcinomas and 38 squamous cell carcinomas. The frequency of metastases increased with local tumor extension (patients with cystectomy: pT2, 36%; pT3a, 45%; pT3b, 69%; pT4, 79%; p < 0.0001). For all pT classifications, the frequency of metastases was slightly higher in patients treated by cystectomy (metastases in 45% of 29 pT2 and 89% of 28 pT4 tumors) than in patients without cystectomy (36% of pT2 and 79% of pT4 tumors). CONCLUSIONS: These results argue against relevant clinical differences between histologic tumor types. The high frequency of metastases in patients having undergone cystectomy indicates that metastasis often occurs before the time of diagnosis. This emphasizes the need for a better prediction of the metastatic capability of these tumors at the time of diagnosis. PMID- 10461107 TI - Interstitial laser coagulation in benign prostatic hyperplasia: A critical evaluation after 2 years of follow-Up. AB - Since 1993 we have prospectively followed a cumulative cohort of males with benign prostatic hyperplasia and symptomatic bladder outlet obstruction who underwent interstitial laser coagulation (ILC) of the prostate. We evaluated the safety and efficacy of ILC with respect to relief of symptoms and bladder outlet obstruction. In addition to the critical evaluation of our clinical results, the perineal and transurethral approaches were investigated as they may make a substantial impact on the overall success rate, including prostate size, number of sticks per prostate volume and type of application. A total of 59 patients were treated with the Nd-YAG laser (mediLas fibertom) between April 1993 and December 1996. At the time of reevaluation, 47 patients had completed a follow-up of up to 24 months. A perineal approach was used in 34%, transurethral in 23%, and a combined approach in 43% of the patients, depending on the preoperative volume of the prostate. 75% were high-risk patients in accordance with the ASA score (ASA III). The efficacy of treatment was assessed 6, 12, 24 and 52 weeks postoperatively in accordance with the International Prostate Symptom Score (IPS/quality of life), cystomanometric studies, peak urinary flow rate, residual volume and volume reduction of the prostate. Reduction of prostatic volume and sticks used per prostate volume were correlated to the overall success rate. A significant improvement in all voiding parameters (flow rate, residual volume), including the symptom score, was observed. P(det) decreased from an average of 90 cm H(2)O preoperatively to 42 cm H(2)O postoperatively after 24 weeks and the mean reduction in prostate volume was 14 cm(3). Interestingly, it was noted that the overall success rate was not size-related. A distinct positive correlation was found in the number of sticks performed and the improvement in objective and subjective parameters. In view of the low morbidity outcome, especially in high risk patients, we proclaim Nd-YAG interstitial laser coagulation of the prostate to be an effective and safe alternative method of treatment for symptomatic benign prostatic hyperplasia. PMID- 10461108 TI - Interaction between osteopontin on madin darby canine kidney cell membrane and calcium oxalate crystal. AB - We recently reported that the addition of the protein osteopontin (OPN) resulted in an increase in the deposition of calcium oxalate (CaOx) crystals on the surface of Madin Darby canine kidney (MDCK) cells. To determine the degree to which this increased deposition is caused by OPN, we investigated the extent to which the CaOx crystal deposition produced by the expression of OPN at the cell surface was suppressed by 4 different methods prior to the determination of the level of CaOx crystal binding. MDCK cells (2 x 10(6) cells/well) were cultured to a confluent state, and the binding of OPN to the cellular surface was then inhibited by adding one of the following 4 substances: human OPN polyclonal antibody, thrombin, cyclic Arg-Gly-Asp (RGD) peptides and tunicamycin. The cells were cultured for 24 h. We then used a fluorescent antibody technique with an OPN polyclonal antibody to determined whether the expression of OPN at the cell surface was inhibited, and we measured the degree of CaOx crystal deposition using the isotope (45)Ca. The degree of CaOx crystal deposition was inhibited by 80% or more in the antibody-treated group, by 50-80% in the thrombin-treated group, by 60-80% in the cyclic RGD-treated group, and by 50-60% in the tunicamycin-treated group. These results suggest that OPN in the extracellular matrix is the main cause of CaOx crystal deposition on the surface of MDCK cells. PMID- 10461109 TI - Excretion of tamm-horsfall protein in patients with uric acid stones. AB - The cause of reduced Tamm-Horsfall protein excretion in patients suffering from uric acid diathesis is still unknown. Our investigation was conducted based on the hypothesis that the solubility of uric acid is increased by Tamm-Horsfall protein and that an increased uric acid content in the urine might cause a decrease in Tamm-Horsfall protein. In 20 patients with uric calculi the excretion of Tamm-Horsfall protein, uric acid, calcium, and citrate was measured. 65% of the patients had pure uric acid stones (group I) and 35% showed mixed stones with at least 30% of uric acid (group II). Reduced Tamm-Horsfall protein excretion was found in 63% of the patients of group I and in 43% of the patients of group II. The excretion of Tamm-Horsfall protein was significantly reduced in pure uric acid stone formers compared to normal subjects (p < 0. 0001). The excretion of uric acid was elevated in 61% of the patients of group I and in 86% of the patients of group II. There was no significant correlation between Tamm-Horsfall protein excretion and uric acid excretion (r = 0.2139). Calcium excretion was elevated in 57% of the patients with mixed stones. The excretion of citrate was reduced in almost all of the patients of groups I and II. Our results do not support the hypothesis that an increased content of uric acid in the urine causes a decrease in Tamm-Horsfall protein. In our opinion the lower excretion of Tamm Horsfall protein in some of the stone patients might be caused by damage in the distal tubular epithelium. Moreover, it has to be supposed that there are defects both in the distal and the proximal tubule in patients prone to develop uric acid calculi. PMID- 10461110 TI - Effect of ageing on blood flow to the bladder and bladder function. AB - Ischemia, induced by atherosclerosis, is a common cause of disorders in the elderly. Bladder dysfunction in older people may be caused by detrusor ischemia. We compared blood flow to the bladder and detrusor function in vivo and in vitro in young (6-month-old) and aged (24-month-old) male Sprague-Dawley rats. In both young and old rats, blood flow to the bladder measured by a laser Doppler flowmeter decreased as intravesical volume increased and was smaller in old rats than in young rats. Cystometrograms performed under anesthesia showed that old rats had smaller voiding pressure and larger bladder capacity than young rats. In isolated bladders, the pressure increase in response to bethanechol and low frequency field stimulation were impaired by aging. Volume-pressure studies showed that in isolated bladders of old rats compliance was greater and peak response to field stimulation was observed at a larger capacity. These findings indicate that bladders of older rats have a larger capacity with good compliance, but less contractility. Aging changes correlate with a decrease in blood flow to the bladder. PMID- 10461112 TI - Paraganglioma of the urinary bladder: A case report and review of the Japanese literature. AB - We describe a patient with primary paraganglioma of the urinary bladder associated with typical clinical symptoms. Systolic blood pressure frequently increased to 300 mm Hg immediately after micturition. Levels of urinary vanillylmandelic acid excretion and serum norepinephrine were significantly elevated. The tumor was removed by partial cystectomy. Histological examination of the tumor revealed paraganglioma of the urinary bladder. Compared with a review of 53 similar cases reported in Japan, the distribution of age, sex and tumor location in the bladder differed from those described worldwide. PMID- 10461111 TI - Management of ureterointestinal obstruction and extensive loss of the ureter by creating a pedicle tube graft from the orthotopic neobladder. AB - We report a case of ureterointestinal obstruction and extensive loss of the ureter following radical cystectomy and orthotopic bladder replacement. The patient was successfully treated by creating a pedicle tube graft from the orthotopic Mainz pouch. PMID- 10461113 TI - Malignant fibrous histiocytoma of the epididymis. Case report and review of the literature. AB - A 54-year-old patient was found to have a painless ca. 1.5 cm diameter tumor in the region of the right epipidymis. The right testis was exposed from a scrotal incision. The final pathology surprisingly showed malignant fibrous histiocytoma (MFH) of the epipidymis and the patient subsequently underwent radical inguinal orchiectomy. Complete metastatic screening was normal. Postoperatively, 60 Gy external beam radiation was administered. At the least follow-up (6 years), there were no recurrences or metastases. MFH is an extremely rare, highly malignant connective tissue tumor, which may, occasionally, affect the male genital tract. There are no agreed treatment principles. The overall prognosis is poor. PMID- 10461114 TI - CT appearance of a renal aspergilloma in a patient with the acquired immunodeficiency syndrome. AB - The appearance of a unilateral renal aspergilloma on computed tomography is described in a 32-year-old man with acquired immunodeficiency syndrome (AIDS). Aspergillus infections are uncommon in the AIDS population. Only 9 cases of renal aspergilloma have been described in AIDS. The treatment performed was percutaneous drainage followed by antifungal drug administration and unilateral nephrectomy. This case report emphasizes the fact that renal fungal infections need to be considered in differential diagnosis of kidney infections in AIDS patients. PMID- 10461115 TI - Stenturia: An unusual manifestation of spontaneous ureteral stent fragmentation. AB - Two patients presented with passage of worm-like stent fragments in the urine. The first had undergone attempted percutaneous removal of left renal calculus and ureteral stenting 4 months prior to presentation. The second had left-sided stent placement for obstructive anuria on account of bilateral renal calculi 3 months earlier. The stents had fragmented into multiple pieces over a mean indwelling time of only 3.5 months. Apart from calculus disease, both patients had documented urinary tract infection. Stent fragmentation is a relatively rare (0.3%) but major complication. However, spontaneous excretion of these fragments has not been hitherto reported. These cases of rapid stent disintegration highlight the need for closer monitoring of the indwelling stents, especially in patients with calculus disease and associated persistent infection. In such patients the stent should probably be changed within 3 months. PMID- 10461116 TI - Vesicocolonic fistula four years after augmentation colocystoplasty. AB - An 8-year-old girl was born with crossed fused renal ectopia and neurogenic bladder due to sacral agenesis. Due to progressive upper tract deterioration and incontinence despite clean intermittent catheterization and pharmacotherapy with anticholinergic agents, the patient underwent augmentation colocystoplasty at the age of 4 years. Four years after surgery the girl was readmitted because of persistent febrile urinary tract infection, persistent metabolic acidosis, and intermittent watery diarrhea. A cystogram revealed a fistula between the dome of the augmented bladder and the transverse colon. The fistula was successfully resected. The presence of enterovesical fistula should always be suspected in a patient with augmented bladder who have late onset of urinary tract infection, metabolic acidosis, and diarrhea. PMID- 10461117 TI - Inflammatory pseudotumor of the urinary bladder and sigmoid colon. AB - We report an unusual case of an inflammatory pseudotumor of the bladder. A biopsy specimen prior to surgical treatment suggested leiomyosarcoma, and computerized tomography and magnetic resonance imaging strongly suggested bladder tumor with sigmoid colon invasion. Accordingly, radical surgical treatment consisting of total cystectomy, anterior resection of the sigmoid colon, and ileal conduit was performed. After operation, however, histological examination revealed an inflammatory pseudotumor of bladder and sigmoid colon. These findings were confirmed by immunohistochemical staining for smooth muscle specific actin, desmin, S-100, and vimentin. Histological examination revealed a diverticulum of the sigmoid colon in the middle of the pseudotumor, and this diverticulum may have ruptured to cause the inflammatory pseudotumor of bladder and sigmoid colon. PMID- 10461118 TI - Unusual semi-spheric perivesical calcification after pelvic radiotherapy. AB - An uncommon case with semi-spheric perivesical calcification after pelvic radiotherapy is reported and the possible pathogenesis of this phenomenon is discussed. PMID- 10461119 TI - Epitheloid sarcoma of the penis. AB - We report a case representing the clinical as well as histopathologic features of epitheloid sarcoma. Both a clinical review and the results of definitive treatment together with the follow-up data are presented. PMID- 10461120 TI - Radio-contrast enhancement of a urinary tract calculus. AB - We report the case of a woman who presented with obstruction and urosepsis of her left kidney secondary to small cystine calculi. The calculi could not be seen on initial plain abdominal X-ray. However, following percutaneous nephrostogram the calculi became more radio-dense and visible on later X-rays. Experimental enhancement of calculi has been described before but not in man. We presume that the contrast medium was adsorbed by the calculus to increase the radio-density. Further investigation of this phenomenon could prove useful in the management of complex radiolucent calculi. PMID- 10461121 TI - An unusual case of ureteral ectopia in the seminal vesicle: diagnosis and surgical management. AB - We report the case of a young man who presented with numerous episodes of ipsilateral epididymitis. Selected imaging studies with consideration of urogenital embryology lead to the rare diagnosis of a dysplastic kidney with ureteral ectopia in the seminal vesicle. After nephroureterectomy and vesiculectomy, convalescence was uneventful. No specific symptoms, equivocal diagnostic findings and the small number of patients limit the surgical experience in diseases of the seminal vesicle. Suprainguinal extravesical extirpation, however, appears to be an excellent operative approach in cases of unilateral seminal vesicle cysts. PMID- 10461122 TI - Deferred imitation by 6- and 9-month-old infants: more evidence for declarative memory. AB - Deferred imitation has recently surfaced as a hallmark measure of nonverbal declarative memory. In two experiments, we examined the developmental origins of deferred imitation during early infancy. Six- and 9-month-old human infants observed an experimenter perform specific actions with multiple objects. The infants' ability to reproduce those actions was assessed following a 24-hr delay. With a single demonstration session, infants of both ages reproduced significantly more actions that had been demonstrated than control actions that had not. These findings challenge the view that memory development is characterized by the emergence of a fundamentally different, declarative memory system later in development. We conclude that the rudiments of declarative memory are present by at least 6 months of age. PMID- 10461123 TI - Long-term maintenance of infant memory. AB - The present experiments with human infants asked whether periodic nonverbal reminders could maintain a memory established at 2 months of age over a substantial period of development. In Experiment 1, a reactivation reminder recovered infants' forgotten memory after 3 weeks, but a reinstatement reminder did not. In Experiment 2, 2-month-olds received a reminder every 3 weeks through 6(1/2) months of age and a final test at 7(1/4) months of age. A preliminary retention test preceded each reminder; which type of reminder (reinstatement or reactivation) infants received depended on performance during this test. Infants exhibited significant retention 4(1/2) months later, and most remembered 5(1/4) months later, when infants outgrew the task. Untrained controls exhibited no retention after any delay. These data confirm that periodic reminders can maintain early memories over significant periods of development and challenge popular claims that preverbal human infants cannot maintain memories over the long term because of neural immaturity or an inability to rehearse experiences by talking about them. PMID- 10461124 TI - Ambient odors associated to failure influence cognitive performance in children. AB - We investigated whether odors can become conditioned to emotionally salient experiences such that when later encountered they influence performance consistent with a previously associated event. To test this hypothesis, 5-year olds were given the experience of failure/frustration on a cognitive maze in a room scented with fragrance and later given another cognitively challenging test in a different room scented with either the same odor, a different odor, or no odor. Results revealed that subjects who performed the test in the presence of the same odor as the maze task did significantly worse than subjects in any other group. Performance in the different odor and the no odor groups were equivalent. Facial expressions and verbal remarks made during the maze task indicated a predominant display of negative affect. These findings show that odors can become conditioned to experiential states and when later encountered have directional influences on behavior. PMID- 10461125 TI - A neural explanation of fetal heart rate patterns: A test of the polyvagal theory. AB - The current study applies a neurophysiological model based on the Polyvagal Theory (Porges, 1995) to interpret fetal heart rate patterns. Beat-to-beat heart rate data from 7 fetuses monitored during the first and second stages of labor were analyzed. Transitory heart rate accelerations and reduced beat-to-beat variability reliably preceded heart rate decelerations. The data are interpreted within the context of the Polyvagal Theory, which provides a plausible explanation of the neurophysiological mechanisms that mediate fetal heart rate decelerations. Specifically, it is proposed that both the transitory heart rate accelerations and the depression of the respiratory rhythm in the beat-to-beat heart rate pattern reflect a withdrawal of the vagal tone determined by myelinated vagal pathways originating in the nucleus ambiguus. Functionally, withdrawal of vagal tone originating in the nucleus ambiguus results in the cardiac pacemaker becoming vulnerable to sympathetic influences and to the more primitive unmyelinated vagal pathways originating in the dorsal motor nucleus of the vagus, which may contribute to clinically relevant bradycardia. PMID- 10461126 TI - Behavioral and psychophysiological correlates of self-presentation in temperamentally shy children. AB - We examined temporal changes in behavior, regional brain electrical activity (EEG), heart rate, cardiac vagal tone, the startle eyeblink response, and salivary cortisol during a task designed to elicit self-presentation anxiety in a group of 7-year-olds, some of whom were classified as temperamentally shy. We found that temperamentally shy children displayed a significantly greater increase in anxious behavior, a greater increase in right, but not left, frontal EEG activity, and a greater increase in heart rate as the task became more demanding compared with their nonshy counterparts. However, the results failed to reveal any significant group differences on the startle eyeblink and salivary cortisol measures. The present findings extend our prior work, in which we found distinct patterns of psychophysiological activity on baseline measures, to differences on psychophysiological measures collected concurrently during a socially evaluative situation in temperamentally shy children. PMID- 10461127 TI - Changes in progressive ratio responding for intravenous cocaine throughout the reproductive process in female rats. AB - Operant responding on a progressive ratio (PR) schedule for intravenous cocaine as well as sucrose reinforcement was examined in female rats throughout the reproductive process. Self-administration sessions began before mating, and continued throughout pregnancy and until lactational Day 8; following parturition, litters were present with dams during operant sessions. Physiological changes associated with the reproductive process dramatically altered PR responding for cocaine, while PR responding for sucrose was relatively stable throughout pregnancy and lactation. Female animals exhibited the highest number of responses/session for cocaine during estrus and the 1st trimester of pregnancy and the lowest responding near parturition, with levels only partially recovering during lactation. Dams self-administering cocaine exhibited notably different patterns of maternal behavior in the operant chambers than dams responding for sucrose. Thus, cocaine's reinforcing efficacy may be influenced by (a) the changing physiological profile associated with the reproductive process and (b) competition from the reinforcing properties of offspring during lactation. PMID- 10461128 TI - Maternal separation disrupts the integrity of the intestinal microflora in infant rhesus monkeys. AB - The integrity of the indigenous microflora of the intestines after maternal separation was investigated in infant rhesus monkeys to determine whether psychological stress may lead to an internal environment conducive to pathogen infection. The stability of the indigenous microflora were estimated by enumeration of total and gram-negative aerobic and facultatively anaerobic bacterial species, specifically Lactobacilli, from coprocultures taken before and after maternal separation. In addition, behavioral and cortisol responses to separation were correlated to the microflora. A significant decrease in fecal bacteria, especially Lactobacilli, was evident on day 3 postseparation, with a return to baseline by the end of the week. The drop in the microflora was correlated with the display of stress-indicative behaviors, but not with cortisol secretion. In addition, infants who displayed numerous stress-indicative behaviors were more susceptible to opportunistic bacterial infection. These results suggest that strong emotional reactions to disruption of the mother infant bond may increase vulnerability to disease. PMID- 10461129 TI - The development of a sex-differentiated defensive motor pattern in rats: A possible role for juvenile experience. AB - When protecting a food item held in the forepaws, rats will dodge laterally away from an approaching conspecific. Both males and females dodge, but do so differently, with females pivoting around the pelvis and males pivoting around the midbody. While females tend to end the dodge with their rumps opposing the other rat's midbody, males typically oppose the head. In this study, two developmental factors were investigated for their role in the genesis of this sex differentiated motor pattern: (a) Dodging by males and females was analyzed before and after puberty to ascertain whether the differential pattern of movement was associated with the pubertal change in pelvic morphology, and (b) Dodging by adult males and females that had been raised without social interaction from weaning was analyzed to ascertain whether experience in the juvenile phase of development was necessary. In both studies, males and females performed the sex-typical version of the dodging motor pattern regardless of age or test condition. However, orienting to the head of the opponent was disrupted in males reared in social isolation, a feature of dodging that developed between weaning and puberty. Therefore, the evidence is consistent with the view that while the sexual differentiation of the motor organization of dodging develops without the need for experience, the males' ability to direct this motor pattern with the correct orientation towards the opponent requires some prepubertal experience. PMID- 10461130 TI - Epidermal glands in Squamata: microscopical examination of precloacal glands in Amphisbaena alba (Amphisbaenia, Amphisbaenidae). AB - The femoral or cloacal region of many species of lizards and amphisbaenians exhibits epidermal glands. The pores of these glands are plugged with holocrine solid secretions that serve as semiochemical sources. Many authors assume that these glands are mainly associated with reproduction and demarcation of territory. The structure of precloacal glands in Amphisbaena alba was previously studied by Antoniazzi et al. (Zoomorphology 113:199-203, 1993; J. Morphol. 221:101-109, 1994). These authors suggested that as the animal moves inside tunnels, the secretion plugs are abraded against the substrate, releasing a secretion trail. Some aspects of the plug were difficult to interpret in fine sections due to the dense and brittle nature of the plug. The morphology of the trail, and the manner of deposition on the substrate, have never been reported. This study presents a primarily scanning electron microscopic description of A. alba precloacal glands and of the secretion plugs. It also demonstrates experimentally the formation of the trail and its fine morphology. The results show that when the plugs scrape against the substrate, their constitution helps them to fragment into tiny pieces, which are spread on the ground, thus forming a trail. Each one of the fragments corresponds to a secretion granule of the precloacal gland's secretory cells. In this way, the trail might have an extensive area for volatilization of semiochemicals, constituting an efficient means of intraspecific communication inside the tunnels. PMID- 10461131 TI - Musculature of the facultative parasite Urastoma cyprinae (Platyhelminthes). AB - In an effort to understand how the feeding motions of Urastoma cyprinae are generated, the arrangement of its musculature was studied using fluorescence microscopy of phalloidin-linked fluorescent stains and conventional light histology and transmission electron microscopy. BODIPY 558/568 phalloidin and Alexa 488 phalloidin resolved a meshwork of ribbon-shaped body-wall muscles as well as inner-body musculature associated with the pharynx and male copulatory organ. The general pattern of body-wall muscles in U. cyprinae is similar to that of other rhabdocoel turbellarians in consisting only of circular, longitudinal, and diagonal fibers; the arrangement of these muscles readily correlates with the bending motions the animal undergoes as it feeds at the surface of gills in bivalves it parasitizes. The orogenital atrium of U. cyprinae lies at the posterior apex of the body, opening at a terminal pore. As evidenced by the arrangement of its epithelium and musculature, it appears to be an invagination of the body wall and comes closest of any such duct studied in turbellarians to satisfying the hypothetical model of a "pseudopharynx," ostensibly adapted as an organ for swallowing and so supplementing the ingestive role of the animal's true pharynx. PMID- 10461132 TI - Testing an inference of function from structure: snake vertebrae do the twist. AB - The zygapophyses and zygosphene-zygantrum articulations of snake vertebrae are hypothesized to restrict or eliminate vertebral torsion. This hypothesis is apparently based solely on the inference of function from structure, despite the limitations of such inferences, as well as contradictory observations and measurements. In this study, I observed and measured axial torsion in gopher snakes, Pituophis melanoleucus. To examine the structural basis of axial torsion, I measured the vertebral articulation angles along the body and the insertion angles of five epaxial muscles. To examine torsion in a natural behavior, I digitized video images and measured the degree of apparent axial torsion during terrestrial lateral undulation. Finally, I measured the mechanical capacity of the vertebral joints for actual torsion over intervals of 10 vertebrae in fresh, skinned segments of the trunk. Vertebral articulation angles vary up to 30 degrees and are associated with variation in torsional capacity along the trunk. The freely crawling P. melanoleucus twisted up to 2.19 degrees per vertebra, which produced substantial overall torsion when added over several vertebrae. The vertebral joints are mechanically capable of torsion up to 2.89 degrees per joint. Therefore, despite the mechanical restriction imposed by the complex articulations, vertebral torsion occurs in snakes and appears to be functionally important in several natural behaviors. Even in cases in which mechanical function appears to be narrowly constrained by morphology, specific functions should not be inferred solely from structural analyses. PMID- 10461133 TI - Role of the midgut gland in purine excretion in the robber crab, Birgus latro (Anomura: Coenobitidae). AB - White fecal strands of Birgus latro are composed of small spherules of uric acid with a mean diameter of 1.6 +/- 0.6 microm. Large numbers of membrane-bound spherules with concentric lamellae are present in the R cells of the midgut gland, so we suggest that lengths of white feces are produced by coordinated secretion of these spherules into the lumen of the midgut gland tubules. There are four cell types in the tubules with embryonic (E) cells at the distal tip, B cells in a narrow band at the distal end and R cells making up the bulk of the tubules and gland. F cells are sparsely scattered among the R cells. Midgut gland tissue was assayed for activities of xanthine dehydrogenase and xanthine oxidase, the two forms of xanthine oxidoreductase. Contrary to previous reports, we found that the midgut gland of B. latro contains only high activities of xanthine dehydrogenase. If proteinase inhibitors were omitted from the assays, however, significant activity of xanthine oxidase was measured, a result we regard as an artifact attributable to the partial conversion of xanthine dehydrogenase to xanthine oxidase by endogenous proteinases. R cells were demonstrated to contain peroxisomes, which may be involved in lipid metabolism rather than synthesis of uric acid. PMID- 10461134 TI - Constructional morphology and mode of attachment of the trunk of Corynosoma cetaceum (Acanthocephala: Polymorphidae). AB - Dead specimens of Corynosoma cetaceum were used to describe the trunk musculature of this species and to infer the use of the trunk as a secondary holdfast. Inferences were based on trunk muscle arrangement, changes in trunk shape, size and distribution of spines, and geometry of tegument thickness. The foretrunk of C. cetaceum is swollen and forms a spiny disk that is bent ventrally. The disk is flattened by several groups of muscles not described previously, which seem able to finely adjust the disk surface over the substratum. Disk attachment appears to be accomplished by two dorsal neck retractor muscles specialized in pulling the anchored proboscis into the foretrunk. This mechanism has been described in other acanthocephalans, becoming surprisingly efficient when used with a flattened, armed foretrunk. The ventrally spined hindtrunk requires force to move downwards in order to attach. A single ventral neck retractor muscle seems specialized in pulling the posterior trunk forward, inducing a downward force due to the muscle's precise points of insertion. This mechanism necessarily generates ventral wrinkling that needs to be eliminated for the spiny surface to be functional. The trunk ventral muscles are apparently arranged so as to concentrate the "excess" of the tegument into a single fold, optimizing the use of the remaining surface for attachment. The size and distribution of spines, as well as the geometry of tegumental thickness, conform to these observations. Morphological changes, seemingly simple, such as structural bending, may have triggered a cascade of subtle modifications and new functions during acanthocephalan evolution, reflecting how morphological integration and novelty interact. PMID- 10461135 TI - Functional and historical determinants of shape in the scapula of Xenarthran mammals: evolution of a complex morphological structure. AB - The mammalian scapula is a complex morphological structure, composed of two ossification plates that fuse into a single structure. Most studies on morphological differentiation in the scapula have considered it to be a simple, spatially integrated structure, primarily influenced by the important locomotor function presented by this element. We used recently developed geometric morphometric techniques to test and quantify functional and phylogenetic influences on scapular shape variation in fossil and extant xenarthran mammals. The order Xenarthra is well represented in the fossil record and presents a stable phylogenetic hypothesis for its genealogical history. In addition, its species present a large variety of locomotor habits. Our results show that approximately half of the shape variation in the scapula is due to phylogenetic heritage. This is contrary to the view that the scapula is influenced only by functional demands. There are large-scale shape transformations that provide biomechanical adaptation for the several habits (arboreality, terrestriality, and digging), and small scale-shape transformations (mostly related to the coracoid process) that are not influenced by function. A nonlinear relationship between morphometric and phylogenetic distances indicates the presence of a complex mixture of evolutionary processes acting on shape differentiation of the scapula. PMID- 10461136 TI - Lateral line morphology and cranial osteology of the rubynose brotula, Cataetyx rubrirostris. AB - Cranial osteology, canal neuromast distribution, superficial neuromast distribution and innervation, and cephalic pore structure were studied in cleared and stained specimens of the deep sea brotulid Cataetyx rubrirostris. The cranial bone structure of C. rubrirostris is similar to other brotulids (Dicrolene sp.) and zoarcids (Zoarces sp.), except for an unusual amount of overlapping of the bones surrounding the cranial vault. The superficial neuromasts are innervated by the anterodorsal, anteroventral, middle and posterior lateral line nerves and are organized similarly to those of the blind ophidioid cave fish Typhliasina pearsei. The cephalic pores open into a widened lateral line canal system. The canal is compartmentalized into a series of neuromast-containing chambers that probably amplify signals received by the system. PMID- 10461138 TI - A message from the editor PMID- 10461137 TI - Muscle development in the abdominal region of larval Hylidae (Amphibia: Anura). AB - Larval muscle development in the abdominal region of five species of hylid frogs (Scinax nasicum, S. fuscovarium, Hyla andina, Phyllomedusa boliviana, Gastrotheca gracilis) was studied using differential staining techniques. These five species represent three major hylid subfamilies. The development of the main abdominal muscles, the rectus abdominis, the two lateral muscles (obliquus externus and transversus), and the lateral pectoralis abdominalis is described. The number of myotomes of the rectus abdominis varies between five and six, and the abdominal muscles associated with the rectus abdominis (obliquus externus, pectoralis abdominalis, and rectus cervicis) vary interspecifically in time of appearance and configuration. The presence of gaps in the configuration of the rectus abdominis has been related to the lotic habits of the larvae. However, our observations indicate the presence of such gaps in larvae that inhabit lentic environments as well. These results suggest that the presence of these gaps is unrelated to larval habitat. There are relatively small differences in muscle morphology among these closely related species, which apparently cannot be explained by morphological adaptations related to their ecology. In the species studied, the number of elements that form the abdominal musculature in larvae is equal to that observed in adults. Likewise, the general morphology of the muscles is ontogenetically conserved. This suggests that both the axial skeleton and musculature are more ontogenetically conserved in relation to the substantial changes that are observed in the skull and head muscles of developing anurans. PMID- 10461145 TI - Discovering ion-channel modulators - making the electrophysiologist's life more interesting. PMID- 10461146 TI - Dizzying but scary: looking towards R&D in 2005. PMID- 10461147 TI - Researchers reveal ways to defeat 'superbugs' PMID- 10461148 TI - Combinatorial chemistry and HTS - feeding a voracious process. PMID- 10461149 TI - Imaging systems in assay screening. AB - High-throughput screening laboratories continuously seek higher throughput, lower cost and compound conservation. In an attempt to approach these goals, improvements in traditional detection instruments (e.g. scintillation counters, fluorescence plate readers, luminometers) are being made. A more radical alternative is the use of image-based detection methods, which have the advantage that large numbers of wells can be quantified in a single detection procedure (favouring miniaturization) and that faint luminescence or scintillation assays can be accomplished very rapidly. However, image-based screening is a very new technology and must prove itself before being broadly accepted. In this review, the technology of a commercial instrument for image-based screening (the LEADseeker from Amersham Pharmacia Biotech, Amersham, UK) is described, and its performance summarized with scintillation, luminescence, fluorescence and absorbance signals. PMID- 10461150 TI - Corrigendum. PMID- 10461151 TI - Microdispensing technologies in drug discovery. AB - Because of the advent of managed care,the pharmaceutical industry is entering a new era, characterized by increased competition and pricing pressures. As a result, drug discovery within pharmaceutical companies is rapidly embracing new paradigms to help bring more novel drugs to the market as rapidly as possible. One paradigm currently being pursued is the miniaturization of the processes involved in the exploratory phase of drug discovery. This reduction in scale has led to the development of new dispensing technologies. This review examines several microdispensing technologies for drug discovery. PMID- 10461153 TI - Correction. PMID- 10461152 TI - RNA as a small-molecule drug target: doubling the value of genomics. AB - Recent advances in the determination of RNA structure and function have led to new opportunities that will have a significant impact on the pharmaceutical industry. RNA, which, among other functions, serves as a messenger between DNA and proteins, was thought to be an entirely flexible molecule without significant structural complexity. However, recent studies have revealed a surprising intricacy in RNA structure. This observation unlocks opportunities for the pharmaceutical industry to target RNA with small molecules. Because both proteins and their specific mRNAs are potential drug-binding sites, the number of targets revealed from genome sequencing efforts is effectively doubled. Perhaps more importantly, drugs that bind to RNA might produce effects that cannot be achieved by drugs that bind to proteins. PMID- 10461154 TI - Cell-based assays and instrumentation for screening ion-channel targets. AB - Ion channels are an important class of drug targets. They comprise the molecular basis for essential physiological functions including fluid secretion, electrolyte balance, bioenergetics and membrane excitability. High-throughput screening for ion-channel function requires sensitive, simple assays and instrumentation that will report ion channel activity in living cells. This article will review relevant assay technologies for ion channels and describe voltage-sensitive probes and instruments based on fluorescence resonance energy transfer (FRET) that enable ion-channel drug discovery. PMID- 10461155 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 10461156 TI - Combinatorial chemistry. PMID- 10461157 TI - Potential use of infrared microspectroscopy to study drug-related structural changes in bone. PMID- 10461158 TI - Echinococcosis from every angle. PMID- 10461159 TI - Praziquantel--its use, cost and possible development of resistance. PMID- 10461160 TI - Transport and trafficking in the malaria-infected erythrocyte. PMID- 10461161 TI - An integrated model of chloroquine action. PMID- 10461162 TI - Na+/H+ antiporter, chloroquine uptake and drug resistance: inconsistencies in a newly proposed model. PMID- 10461164 TI - Websites of interest PMID- 10461163 TI - Beaver fever and pinworm neuroses on the Net. PMID- 10461165 TI - GTPases in protozoan parasites: tools for cell biology and chemotherapy. AB - Small G proteins belong to a superfamily of GTPases related to the protooncogene ras, and function as master control elements for a range of cellular functions. This ability is related to their low rate of substrate turnover; GTPases catalyse the conversion of GTP to GDP, but with a rate in the order of one substrate per second, orders of magnitude slower than 'good' enzyme catalysis, but placing the reaction into the temporal frame of many cellular processes including signal transduction, cytoskeletal reorganization and vesicle trafficking. In this article, Mark Field, Bassam Ali and Helen Field describe some recent advances in G-protein studies in the parasite field, concentrating on the protozoan parasites. Because of their numerous roles in cell biology, understanding parasite G proteins has great potential for increasing our knowledge of parasite cellular physiology, as well as providing important inroads into vital processes for potential therapeutic exploitation. PMID- 10461166 TI - Genetic manipulation of kinetoplastida. AB - During the 1980s, many kinetoplastid genes were cloned and their function inferred from homology with genes from other organisms, location of the corresponding proteins or expression in heterologous systems. Up until 1990, before the availability of DNA transfection methodology, we could not analyze the function of kinetoplastid genes within the organisms themselves. Since then, it has become possible to create and complement mutants, to overexpress foreign proteins in the parasites, to knock out genes and even to switch off essential functions. However, these methods are not equally applicable in all parasites. Here, Christine Clayton highlights the differences and similarities between the most commonly used model organisms, and assesses the relative advantages of different approaches and parasites for different types of investigation. PMID- 10461167 TI - Schistosomiasis of the female genital tract: public health aspects. AB - In this paper Gabriele Poggensee, Hermann Feldmeier and Ingela Krantz discuss the public health relevance of female genital schistosomiasis (FGS). Some of the stated hypotheses are supported only by clinical observations and/or circumstantial evidence as valid epidemiological and immunological data of this disease entity are still very scanty. Morbidity caused by the presence of schistosome eggs in the lower and upper genital tract have been almost completely neglected during the past two decades. This has been acknowledged by the WHO and, in 1997, the Gender Task Force of the WHO's Tropical Disease Research Programme (TDR) included FGS in a list of scientific areas that deserve high research priority. PMID- 10461168 TI - The role of albendazole in programmes to eliminate lymphatic filariasis. AB - Citing earlier advances in the treatment of lymphatic filariasis [particularly the effectiveness of single-dose diethylcarbamazine (DEC) in reducing microfilaraemia and its enhanced effectiveness when co-administered with single dose ivermectin], Eric Ottesen, Mahroof Ismail and John Horton consider recent studies on the antifilarial activity of albendazole that have led to the current recommendations for its use in single-dose regimens in conjunction with either DEC or ivermectin for large-scale control/elimination programmes. Furthermore, the potential of albendazole as a macrofilaricide for treating individual patients with lymphatic filarial infections is emphasized as one of a number of important research questions that remain to be explored. PMID- 10461169 TI - Human dirofilariasis in the European Union. AB - The dog parasites Dirofilaria immitis and D. (Nochtiella) repens, well known as zoonotic agents, are widely distributed in southern Europe. Although both species are canine parasites, infection with immature worms has been found in humans, who develop nodules, mainly in subcutaneous tissue or in lung parenchyma arising from branches of the pulmonary artery. In humans, the parasites do not usually reach the adult stage and microfilaremia is absent, as has been shown by diagnosis using invasive methods for removing the nodules. In this article, Antonio Muro, Claudio Genchi, Miguel Cordero and Fernando Simon review the current situation concerning the clinical and epidemiological aspects, immune response and diagnosis of human dirofilariases. PMID- 10461170 TI - The strains of retyping Leishmania. PMID- 10461171 TI - Crosstalk Among Multiple Signaling Pathways Controlling Ovarian Cell Death. AB - Ovarian cell death is an essential process for the homeostasis of ovarian function in human and other mammalian species. It ensures the selection of the dominant follicle and the demise of excess follicles. In turn, this process minimizes the possibility of multiple embryo development during pregnancy and assures the development of few but healthy embryos. Degeneration of the old corpora lutea in each estrous/menstrual cycle by programmed cell death is essential to maintain the normal cyclicity of ovarian steroidogenesis. Although there are multiple pathways that can determine cell death or survival, crosstalk among endocrine, paracrine and autocrine factors, as well as among protooncogenes, tumor suppressor genes, survival genes and death genes, plays an important role in determining the fate of ovarian somatic and germ cells. The establishment of immortalized rat and human steroidogenic granulosa cell lines and the investigation of pure populations of primary granulosa cells allows systematic studies of the mechanisms that control steroidogenesis and apoptosis of granulosa cells. These cells are the most abundant type of somatic follicular cell. Moreover, crosstalk between p53 and extracellular matrix components such as laminin, fibronectin and basic fibroblast growth factor, between cAMP- and p53 generated signals and between steroid hormones and Bcl-2, can explain some of the fine tuning that controls ovarian steroidogenesis and apoptosis. Further study of the mechanisms of ovarian cell death will lead to a better understanding of the processes involved and permit the formulation of novel strategies for the treatment of ovarian malfunctions, such as polycystic ovarian syndrome and ovarian cancer. PMID- 10461172 TI - The PTB Domain: The Name Doesn't Say It All. AB - The phosphotyrosine-binding domain is a recently described protein-protein interaction domain which, despite its name, is involved in both phosphotyrosine dependent and -independent interactions. Proteins with this domain are involved in diverse cellular functions, ranging from receptor signaling to protein targeting. PMID- 10461173 TI - Pathophysiological Role of Growth Factors in Diabetic Kidney Disease: Focus on Innovative Therapy. AB - Various growth factors have been proposed to be players in the development of diabetic microvascular complications. In particular, the growth hormone/insulin like growth factor system and the transforming growth factor beta system have measurable effects on the development of diabetic kidney disease through a complex intrarenal system. Recent findings indicating that these growth factors might be responsible for early renal changes in diabetes have provided insight into processes that might be relevant to the future development of new drugs useful in the treatment of diabetic kidney disease. PMID- 10461174 TI - Neurohormonal Signaling Pathways and the Regulation of Alzheimer beta-Amyloid Precursor Metabolism. AB - Postmenopausal women on estrogen replacement appear to have their relative risk of developing Alzheimer's disease diminished by about one half. Because brain amyloid accumulation plays a key role in initiating Alzheimer's pathology, it is attractive to postulate that estrogen might modulate Alzheimer's risk by inhibiting amyloid accumulation. Data and cell biological models supporting such a scenario are reviewed here. PMID- 10461175 TI - Genetic Analysis in Fine-needle Aspiration of the Thyroid: A New Tool for the Clinic. AB - This review will examine the application of genetic analysis to cytological specimens obtained by fine-needle aspiration biopsy in the management of thyroid tumours. In particular, it will consider the use of polymerase chain reaction based techniques in the diagnosis of the micrometastasis of differentiated thyroid carcinoma to cervical lymph nodes, as well as in the detection of oncogenic alterations in solid thyroid nodules. PMID- 10461176 TI - Why did biocatalysis in organic media not take off in the 1930s? PMID- 10461177 TI - A general strategy for enzyme engineering. PMID- 10461178 TI - A guide to the WWW PMID- 10461179 TI - A spatial approach to transcriptional profiling: mechanotransduction and the focal origin of atherosclerosis. AB - The initiation and progression of focal atherosclerotic lesions has long been known to be associated with regions of disturbed blood flow. Improved precision in experimental models of spatially defined flow has recently been combined with regional and single-cell gene-expression profiling to investigate the relationships linking haemodynamics to vessel-wall pathobiology. PMID- 10461180 TI - Kleisli: a new tool for data integration in biology. AB - One of the central problems in bioinformatics is data retrieval and integration. The existing biological databases are geographically distributed across the Internet, complex and heterogeneous in data types and data structures, and constantly changing. With the current rapid growth of biomedical data, the challenge is how large volumes of data retrieved from multiple databases can be transformed and integrated automatically and flexibly. This article describes a powerful new tool, the Kleisli system, for complex queries across multiple databases and data integration. PMID- 10461181 TI - Improving nutrient capture from soil by the genetic manipulation of crop plants AB - Modern agricultural practices require the massive application of fertilizer to soils worldwide. In the USA alone, millions of tons of fertilizer are applied to agricultural soils each year. The adverse environmental impact and high cost of fertilizer use underscore the importance of improving the capability of plants to capture nutrients from soil. Therefore, engineering plants with improved micronutrient uptake may help in creating sustainable agriculture in the next century. PMID- 10461182 TI - Tandem constructs: preventing the rise of superweeds. AB - Transgenic crops may interbreed with nearby weeds, increasing their competitiveness, and may themselves become a 'volunteer' weed in the following crop. The desired transgene can be coupled in tandem with genes that would render hybrid offspring or volunteer weeds less able to compete with crops, weeds and wild species. Genes that prevent seed shatter or secondary dormancy, or that dwarf the recipient could all be useful for mitigation and may have value to the crop. Many such genes have been isolated in the past few years. PMID- 10461183 TI - Biopharmaceutical production in transgenic livestock. AB - The production of recombinant human proteins in the milk of transgenic dairy animals offers a safe, renewable source of commercially important proteins that cannot be produced as efficiently in adequate quantities by other methods. A decade of success in expressing a variety of proteins in livestock has brought three human recombinant proteins to human clinical trials. Recent progress has drawn on molecular biology and reproductive physiology to improve the efficiency of producing and reproducing useful transgenic founder animals, and to improve the expression of heterologous proteins in their milk. PMID- 10461184 TI - Prospects for drug screening using the reverse two-hybrid system. AB - Rational drug-screening strategies have been limited by the number of available protein targets. The fields of genomics and functional genomics are now merging into 'chemical genomics' approaches, in which large numbers of potential target proteins can be used in standardized high-throughput drug-screening assays. Because protein-protein interactions are critical to most biological processes and can be tested in standardized assays, they may represent optimal targets in the chemical-genomics era. The reverse two-hybrid system appears to have several properties that would be critical for the success of this approach. PMID- 10461185 TI - Clathrin: anatomy of a coat protein. AB - Clathrin is a vesicle coat protein involved in the assembly of membrane and cargo into transport vesicles at the plasma membrane and on certain intracellular organelles. Recently, crystal structures of two separate parts of the clathrin heavy chain, a fragment of the proximal leg and the N-terminal domain, have been analysed, providing the first high-resolution data for a vesicle coat protein. Viewing these structures in the context of a hexagonal barrel coat, recently determined to 21 A by cryo-electron microscopy, provides new insights into the assembly of clathrin coats. PMID- 10461186 TI - Gamma-tubulin complexes: size does matter. AB - gamma-Tubulin is a conserved component of all microtubule-organizing centres and is required for these organelles to nucleate microtubule polymerization. However, the mechanism of nucleation is not known. In addition to its localization to organizing centres, a large pool of gamma-tubulin exists in the cytoplasm in a complex with other proteins. The size of the gamma-tubulin complex and number of associated proteins vary among organisms, and the functional significance of these differences is unknown. Recently, the nature of these gamma-tubulin complexes has been explored in different organisms, and this has led us closer to a molecular understanding of microtubule nucleation. PMID- 10461187 TI - The coronin family of actin-associated proteins. AB - Coronin was first isolated from Dictyostelium, but similar proteins have been identified in many species and individual cell types. The coronin-like protein in yeast promotes actin polymerization and also interacts with microtubules. Dictyostelium mutants lacking coronin are impaired in cytokinesis and all actin mediated processes. Analysis of coronin-GFP (green-fluorescent protein) fusions and knockout mutants shows that coronin participates in the remodelling of the cortical actin cytoskeleton that is responsible for phagocytosis and macropinocytosis. Likewise, in mammalian neutrophils, a coronin-like protein is also associated with the phagocytic apparatus. The diversity of function in this family of actin-associated proteins is just beginning to be explored. PMID- 10461188 TI - Pak to the future. AB - Members of the Pak family of serine/threonine kinases serve as targets for the small GTP-binding proteins Cdc42 and Rac and have been implicated in a wide range of biological activities. Recently, some exciting developments help elaborate the regulation of Pak activity and identify downstream signalling targets. These include the discovery of the Cool/Pix and Cat proteins, which modulate Pak signalling, and downstream kinases that modulate the organization of the actin cytoskeleton or gene expression. We present these recent findings and consider how these new regulators and targets could explain some of the cellular effects that have been attributed to Pak family members. PMID- 10461189 TI - Membrane trafficking of neurotransmitter transporters in the regulation of synaptic transmission. AB - Many psychoactive drugs influence the transport of neurotransmitters across biological membranes, suggesting that the physiological regulation of neurotransmitter transport might contribute to normal and perhaps abnormal behaviour. Over the past few years, molecular characterization of the neurotransmitter transporters has enabled investigation of their subcellular location and regulation. The analysis of location suggests that membrane trafficking has an important role in the normal function of these proteins. One of the major regulatory mechanisms also involves changes in localization that might contribute to synaptic plasticity. This article discusses recent work on the membrane trafficking of neurotransmitter transporters and its role in regulating their activity. PMID- 10461191 TI - Careers-perspective interview. PMID- 10461190 TI - Putting a new twist on actin: ADF/cofilins modulate actin dynamics. AB - The actin-depolymerizing factor (ADF)/cofilins are a family of essential actin regulatory proteins, ubiquitous among eukaryotes, that enhance the turnover of actin by regulating the rate constants of polymerization and depolymerization at filament ends, changing the twist of the filament and severing actin filaments. Genetic and cell-biological studies have shown that an ADF/cofilin is required to drive the high turnover of the actin cytoskeleton observed in vivo. The activity of ADF/cofilin is regulated by a variety of mechanisms, including specific phosphorylation and dephosphorylation. This review addresses aspects of ADF/cofilin structure, dynamics, regulation and function. PMID- 10461193 TI - Corrigendum. PMID- 10461192 TI - When are concepts one or many? PMID- 10461194 TI - How infants begin to extract words from speech. AB - A crucial step for acquiring a native language vocabulary is the ability to segment words from fluent speech. English-learning infants first display some ability to segment words at about 7.5 months of age. However, their initial attempts at segmenting words only approximate those of fluent speakers of the language. In particular, 7.5-month-old infants are able to segment words that conform to the predominant stress pattern of English words. The ability to segment words with other stress patterns appears to require the use of other sources of information about word boundaries. By 10.5 months, English learners display sensitivity to additional cues to word boundaries such as statistical regularities, allophonic cues and phonotactic patterns. Infants' word segmentation abilities undergo further development during their second year when they begin to link sound patterns with particular meanings. By 24 months, the speed and accuracy with which infants recognize words in fluent speech is similar to that of native adult listeners. This review describes how infants use multiple sources of information to locate word boundaries in fluent speech, thereby laying the foundations for language understanding. PMID- 10461195 TI - Perception of self-motion from visual flow. AB - Accurate and efficient control of self-motion is an important requirement for our daily behavior. Visual feedback about self-motion is provided by optic flow. Optic flow can be used to estimate the direction of self-motion ('heading') rapidly and efficiently. Analysis of oculomotor behavior reveals that eye movements usually accompany self-motion. Such eye movements introduce additional retinal image motion so that the flow pattern on the retina usually consists of a combination of self-movement and eye movement components. The question of whether this 'retinal flow' alone allows the brain to estimate heading, or whether an additional 'extraretinal' eye movement signal is needed, has been controversial. This article reviews recent studies that suggest that heading can be estimated visually but extraretinal signals are used to disambiguate problematic situations. The dorsal stream of primate cortex contains motion processing areas that are selective for optic flow and self-motion. Models that link the properties of neurons in these areas to the properties of heading perception suggest possible underlying mechanisms of the visual perception of self-motion. PMID- 10461196 TI - Development of theory of mind and executive control. AB - Several recent studies have demonstrated a developmental link, in the age range of 3-5 years, between the acquisition of a 'theory of mind' and self control. In this review, we consider the existence of such a link in assessing five competing theoretical hypotheses that might help us to understand the nature of this developmental advance: (1) executive control depends on theory of mind; (2) theory of mind development depends on executive control; (3) the relevant theory of mind tasks require executive control; (4) both kinds of task require the same kind of embedded conditional reasoning; (5) theory of mind and executive control involve the same brain region. We briefly describe these theoretical accounts and evaluate them in the light of existing empirical evidence. At present, only account (3) can be ruled out with some confidence. PMID- 10461197 TI - An embodied cognitive science? AB - The last ten years have seen an increasing interest, within cognitive science, in issues concerning the physical body, the local environment, and the complex interplay between neural systems and the wider world in which they function. Yet many unanswered questions remain, and the shape of a genuinely physically embodied, environmentally embedded science of the mind is still unclear. In this article I will raise a number of critical questions concerning the nature and scope of this approach, drawing a distinction between two kinds of appeal to embodiment: (1) 'Simple' cases, in which bodily and environmental properties merely constrain accounts that retain the focus on inner organization and processing; and (2) More radical appeals, in which attention to bodily and environmental features is meant to transform both the subject matter and the theoretical framework of cognitive science. PMID- 10461198 TI - Neuromodulation: acetylcholine and memory consolidation. AB - Clinical and experimental evidence suggests that hippocampal damage causes more severe disruption of episodic memories if those memories were encoded in the recent rather than the more distant past. This decrease in sensitivity to damage over time might reflect the formation of multiple traces within the hippocampus itself, or the formation of additional associative links in entorhinal and association cortices. Physiological evidence also supports a two-stage model of the encoding process in which the initial encoding occurs during active waking and deeper consolidation occurs via the formation of additional memory traces during quiet waking or slow-wave sleep. In this article I will describe the changes in cholinergic tone within the hippocampus in different stages of the sleep-wake cycle and will propose that these changes modulate different stages of memory formation. In particular, I will suggest that the high levels of acetylcholine that are present during active waking might set the appropriate dynamics for encoding new information in the hippocampus, by partially suppressing excitatory feedback connections and so facilitating encoding without interference from previously stored information. By contrast, the lower levels of acetylcholine that are present during quiet waking and slow-wave sleep might release this suppression and thereby allow a stronger spread of activity within the hippocampus itself and from the hippocampus to the entorhinal cortex, thus facilitating the process of consolidation of separate memory traces. PMID- 10461199 TI - Multi-organellar disorders of pigmentation: intracellular traffic jams in mammals, flies and yeast. AB - Several different mutant genes in humans, mice and Drosophila, most of which were identified initially on the basis of reduced pigmentation, have been associated with defects of multiple cytoplasmic organelles - melanosomes, lysosomes and granules. Recent discoveries show that several of these mutations directly affect components in the pathway of organelle-specific protein trafficking, and provide new insights into the relationships of these pathways in mammals, flies and yeast. PMID- 10461200 TI - The best yeast? PMID- 10461201 TI - Gene-cluster analysis in chloroplast genomics. PMID- 10461202 TI - Evolution of a conserved protein module from Archaea to plants. PMID- 10461203 TI - Bayesian statistics in genetics: a guide for the uninitiated. AB - Statistical analyses are used in many fields of genetic research. Most geneticists are taught classical statistics, which includes hypothesis testing, estimation and the construction of confidence intervals; this framework has proved more than satisfactory in many ways. What does a Bayesian framework have to offer geneticists? Its utility lies in offering a more direct approach to some questions and the incorporation of prior information. It can also provide a more straightforward interpretation of results. The utility of a Bayesian perspective, especially for complex problems, is becoming increasingly clear to the statistics community; geneticists are also finding this framework useful and are increasingly utilizing the power of this approach. PMID- 10461204 TI - RNA-triggered gene silencing. AB - Double-stranded RNA (dsRNA) has recently been shown to trigger sequence-specific gene silencing in a wide variety of organisms, including nematodes, plants, trypanosomes, fruit flies and planaria; meanwhile an as yet uncharacterized RNA trigger has been shown to induce DNA methylation in several different plant systems. In addition to providing a surprisingly effective set of tools to interfere selectively with gene function, these observations are spurring new inquiries to understand RNA-triggered genetic-control mechanisms and their biological roles. PMID- 10461205 TI - Why have organelles retained genomes? AB - The observation that chloroplasts and mitochondria have retained relics of eubacterial genomes and a protein-synthesizing machinery has long puzzled biologists. If most genes have been transferred from organelles to the nucleus during evolution, why not all? What selective pressure maintains genomes in organelles? Electron transport through the photosynthetic and respiratory membranes is a powerful - but dangerous - source of energy. Recent evidence suggests that organelle genomes have persisted because structural proteins that maintain redox balance within bioenergetic membranes must be synthesized when and where they are needed, to counteract the potentially deadly side effects of ATP generating electron transport. PMID- 10461206 TI - Pax 6: mastering eye morphogenesis and eye evolution. AB - Pax 6 genes from various animal phyla are capable of inducing ectopic eye development, indicating that Pax 6 is a master control gene for eye morphogenesis. It is proposed that the various eye-types found in metazoa are derived from a common prototype, monophyletically, by a mechanism called intercalary evolution. PMID- 10461207 TI - Internet resources for the functional analysis of 5' and 3' untranslated regions of eukaryotic mRNAs. PMID- 10461208 TI - It's a knockout! AB - 'It's a Knockout!' provides an update of some of the latest mouse knockouts in TBASE (http://www.jax.org/tbase/ and Ref. 1). The column provides a concise phenotypic profile of novel mutants and renders their complete characterization directly accessible to Web users, via unique and unchanging accession numbers (TBASE identities). Where possible, interesting knockouts will be grouped according to gene families, application or phenotypic similarities. PMID- 10461209 TI - The scramble to patent human genes. PMID- 10461210 TI - Conscious awareness, memory and the hippocampus. AB - Declarative memory, the conscious recollection of past experiences, is known to involve the hippocampus. Now a study of amnesic patients shows that hippocampus dependent learning can occur in the absence of conscious awareness. PMID- 10461211 TI - Semaphorins: repulsive guidance molecules show their attractive side. AB - Semaphorins are known to repel growth cones of developing axons, but a study of the grasshopper limb bud shows that they can also serve as attractive guidance cues. PMID- 10461212 TI - A sonic boom for gene delivery. AB - A new method for infecting embryos with retroviral vectors allows transgenes to be expressed with high efficiency during very early stages of neural development. PMID- 10461214 TI - Synaptic physiology in C. elegans PMID- 10461213 TI - Anxiety: at the intersection of genes and experience. AB - Human anxiety disorders arise from a combination of genetic vulnerability and traumatic experience. Mice with a GABAA receptor mutation may provide a model for these disorders. PMID- 10461215 TI - "Both" means more than "two": localizing and counting in patients with visuospatial neglect. PMID- 10461216 TI - Calcium channel activation stabilizes a neuronal calcium channel mRNA. AB - We have identified a calcium-dependent pathway in neurons that regulates expression levels of the alpha1B subunit and N channel current. When neurons are depolarized and voltage-gated calcium channels activated, the half-life of cellular N channel alpha1B mRNA is prolonged. This stabilizing effect of depolarization is mediated through the 3' untranslated region of a long form of the alpha1B mRNA and may represent a form of modulation of N-channel levels that does not require changes in gene transcription. Increases in N channel expression would affect several key neuronal functions controlled by calcium, including transmitter release and neurite outgrowth. PMID- 10461217 TI - One GABA and two acetylcholine receptors function at the C. elegans neuromuscular junction. AB - We describe an electrophysiological preparation of the neuromuscular junction of the nematode C. elegans, which adds to its considerable genetic and genomic resources. Mutant analysis, pharmacology and patch-clamp recording showed that the body wall muscles of wild-type animals expressed a GABA receptor and two acetylcholine receptors. The muscle GABA response was abolished in animals lacking the GABA receptor gene unc-49. One acetylcholine receptor was activated by the nematocide levamisole. This response was eliminated in mutants lacking either the unc-38 or unc-29 genes, which encode alpha and non-alpha acetylcholine receptor subunits, respectively. The second, previously undescribed, acetylcholine receptor was activated by nicotine, desensitized rapidly and was selectively blocked by dihydro-beta-erythroidine, thus explaining the residual motility of unc-38 and unc-29 mutants. By recording spontaneous endogenous currents and selectively eliminating each of these receptors, we demonstrated that all three receptor types function at neuromuscular synapses. PMID- 10461218 TI - Ectopic semaphorin-1a functions as an attractive guidance cue for developing peripheral neurons. AB - Transmembrane and secreted glycoproteins of the semaphorin family are typically classified as inhibitory neuronal guidance molecules. However, although chemorepulsive activity has been demonstrated for several semaphorin family members, little is known about the function of the numerous transmembrane semaphorins identified to date. Here we demonstrated that the extracellular semaphorin domain of a transmembrane semaphorin, semaphorin-1a, could actively perturb axon pathfinding in vivo when presented homogenously as a recombinant freely soluble factor. When ectopic overexpression was limited to defined epithelial regions, semaphorin-1a could directly steer axons by acting as an attractive guidance molecule. PMID- 10461219 TI - Continual remodeling of postsynaptic density and its regulation by synaptic activity. AB - A postsynaptic density (PSD) protein, PSD-95, was tagged with green fluorescent protein (GFP-PSD-95) and expressed in cultured hippocampal neurons using recombinant adenoviruses. GFP-PSD-95 was selectively localized to excitatory postsynaptic sites. Time-lapse fluorescence imaging of hippocampal neurons revealed that >20% of GFP-PSD-95 clusters turned over within 24 hours. The appearance rate of clusters was higher than the disappearance rate, and this difference accounted for the gradual increase of the cluster density observed in culture. Dynamics of PSD-95 clusters were also inhibited by blockers of excitatory synaptic transmission. Continual PSD turnover and its regulation by synaptic activity may be important in activity-dependent remodeling of neuronal connections. PMID- 10461220 TI - A method for rapid gain-of-function studies in the mouse embryonic nervous system. AB - We used ultrasound image-guided injections of high-titer retroviral vectors to obtain widespread introduction of genes into the mouse nervous system in utero as early as embryonic day 8.5 (E8.5). The vectors used included internal promoters that substantially improved proviral gene expression in the ventricular zone of the brain. To demonstrate the utility of this system, we extended our previous work in vitro by infecting the telencephalon in vivo as early as E8. 5 with a virus expressing Sonic Hedgehog. Infected embryos showed gross morphological brain defects, as well as ectopic expression of ventral telencephalic markers characteristic of either the medial or lateral ganglionic eminences. PMID- 10461221 TI - Dendritic asymmetry cannot account for directional responses of neurons in visual cortex. AB - A simple model was proposed to account for the direction selectivity of neurons in the primary visual cortex, area V1. In this model, the temporal asymmetries in the summation of inhibition and excitation that produce directionality were generated by structural asymmetries in the tangential organization of the basal dendritic tree of cortical neurons. We reconstructed dendritic trees of neurons with known direction preferences and found no correlation between the small biases of a neuron's dendritic morphology and its direction preference. Detailed simulations indicated that even when the electrotonic asymmetries in the dendrites were extreme, as in cortical Meynert cells, the biophysical properties of single neurons could contribute only partially to the directionality of cortical neurons. PMID- 10461222 TI - Receptive fields of disparity-selective neurons in macaque striate cortex. AB - To identify neuronal mechanisms underlying stereopsis, we characterized interactions between inputs from the two eyes in disparity-selective neurons in macaque V1. All disparity-selective cells showed suppressive interactions between the right and left eyes, and some showed facilitatory interactions. Disparity selectivity was narrower than the receptive-field width and was constant across the receptive field. Such position-invariant disparity selectivity is also found in anesthetized cat V1. Complex cells have been suggested to inherit their disparity selectivity from simple cells with receptive fields mismatched between the two eyes. However, we found no such antecedent disparity-tuned simple cells. We did find disparity-selective cells with some simple-cell characteristics, but surprisingly, they also showed position-invariant disparity selectivity rather than simple linear binocular interactions. PMID- 10461223 TI - Decreased GABAA-receptor clustering results in enhanced anxiety and a bias for threat cues. AB - Patients with panic disorders show a deficit of GABAA receptors in the hippocampus, parahippocampus and orbitofrontal cortex. Synaptic clustering of GABAA receptors in mice heterozygous for the gamma2 subunit was reduced, mainly in hippocampus and cerebral cortex. The gamma2 +/- mice showed enhanced behavioral inhibition toward natural aversive stimuli and heightened responsiveness in trace fear conditioning and ambiguous cue discrimination learning. Implicit and spatial memory as well as long-term potentiation in hippocampus were unchanged. Thus gamma2 +/- mice represent a model of anxiety characterized by harm avoidance behavior and an explicit memory bias for threat cues, resulting in heightened sensitivity to negative associations. This model implicates GABAA-receptor dysfunction in patients as a causal predisposition to anxiety disorders. PMID- 10461224 TI - Stereoscopic occlusion junctions. AB - Portions of surfaces in a binocularly viewed scene may be 'half occluded', that is, visible in only one eye. The human visual system uses zones of half occlusion to help segment the visual scene and infer figure-ground relationships at object boundaries. We developed a quantitative model of the depth-discontinuity cue provided by half occlusion. Half occlusions are revealed by two-dimensional interocular displacements of binocularly viewed occlusion junctions, such as T junctions. We derived a formula relating this two-dimensional displacement, or 'pseudodisparity', to binocular disparities and orientations of occluding and occluded contours. In human psychophysical experiments, perceived depth and contour orientation quantitatively depended on pseudodisparity, as predicted by our model, implying that the visual system senses quantitative variations in interocular junction position to reconstruct occlusion geometry. PMID- 10461226 TI - SOD1 mutants linked to amyotrophic lateral sclerosis selectively inactivate a glial glutamate transporter PMID- 10461227 TI - Reply PMID- 10461225 TI - Memory deficits for implicit contextual information in amnesic subjects with hippocampal damage. AB - The role of the hippocampus and adjacent medial temporal lobe structures in memory systems has long been debated. Here we show in humans that these neural structures are important for encoding implicit contextual information from the environment. We used a contextual cuing task in which repeated visual context facilitates visual search for embedded target objects. An important feature of our task is that memory traces for contextual information were not accessible to conscious awareness, and hence could be classified as implicit. Amnesic patients with medial temporal system damage showed normal implicit perceptual/skill learning but were impaired on implicit contextual learning. Our results demonstrate that the human medial temporal memory system is important for learning contextual information, which requires the binding of multiple cues. PMID- 10461228 TI - Immunocytochemistry and the diagnosis of schistosomiasis: ancient and modern. PMID- 10461229 TI - Changes in cerebral cortex size are governed by fibroblast growth factor during embryogenesis PMID- 10461230 TI - Signaling myopia PMID- 10461231 TI - Dendritic Ih normalizes temporal summation in hippocampal CA1 neurons PMID- 10461232 TI - Timing of cochlear feedback: spatial and temporal representation of a tone across the basilar membrane PMID- 10461233 TI - Future directions in tissue heart valves: impact of recent insights from biology and pathology. PMID- 10461234 TI - Mitral valve replacement with a pulmonary autograft: initial experience. AB - BACKGROUND AND AIM OF THE STUDY: For long-term substitution of the mitral valve, mechanical prostheses require life-long anticoagulation which is impractical in developing countries, xenografts degenerate early in our young population, and mitral homografts have not yet been established as being suitable. We therefore returned to an original concept first reported by one of the authors (D.N.R.) in 1967. METHODS: Between July 1997 and November 1998, 22 patients (mean age 40.3 years; range: 28 to 57 years) with rheumatic mitral valve disease unsuitable for reconstruction were subjected to excision of their pulmonary valve in the standard fashion of the Ross procedure. The inverted autograft was incorporated in a 2.5 cm-long Dacron conduit, with a pericardial collar attached to its proximal end. The distal end of the autograft-conduit was sutured to the annulus of the excised mitral valve, and the proximal end incorporating the pericardial collar was attached to the adjacent atrial wall. In this way all prosthetic material was covered. The right ventricular outflow was reconstructed with a pulmonary homograft in 17 patients, with an aortic homograft in two, and with a porcine pulmonary xenograft in three. RESULTS: One patient developed a fatal cerebrovascular accident, probably related to an incorrectly placed pericardial collar with rough surface exposed to the blood flow. In a second patient the autograft had to be replaced six weeks after operation due to bacterial endocarditis contracted in the operating room. Echocardiography confirmed excellent function of the remaining autografts up to 16 months postoperatively (mean follow up 8.3 months). CONCLUSIONS: We believe the pulmonary autograft to be a valid option for mitral valve replacement in our patients. PMID- 10461235 TI - Lack of durability of the Mitroflow valve does not affect survival. AB - BACKGROUND AND AIM OF THE STUDY: The aim of the study was to compare the durability and risk of reoperation in patients undergoing isolated aortic valve replacement with either a porcine standard Carpentier Edwards or a Mitroflow pericardial valve. METHODS: Follow up evaluation was performed in 118 patients receiving a Mitroflow valve (M) and 94 patients receiving a standard Carpentier Edwards porcine valve (CE) between 1980 and 1987. The two groups were identical in terms of clinical characteristics; only prosthesis size differed, with small sized valves used more frequently among the Mitroflow group. RESULTS: The risk of structural valve deterioration (SVD) was 2.3% per patient-year (pt-yr) in CE valves, and 5.4 per pt-yr in M valves. Freedom from SVD was 100%, 87 +/- 4% and 63 +/- 8% at 5, 10 and 15 years for CE valves, and 96 +/- 2%, 56 +/- 7% and 5 +/- 4% for M valves. Freedom of reoperation was 98 +/- 1%, 83 +/- 5% and 76 +/- 7% at 5, 10 and 15 years respectively for CE valves, and 94 +/- 2%, 55 +/- 7% and 11 +/ 9% for M valves. Despite the high number of valve-related reoperations, survival at 5, 10 and 15 years was not affected in M valve patients. Multivariate analysis (Cox model) showed that age and valve type were the two main risk factors for SVD and reoperation, though the latter factor had no impact on survival. CONCLUSIONS: In younger patients (aged < 75 years), the CE valves offer superior results to the M counterpart in valve replacement. However, in patients aged > 75 years, pericardial and porcine bioprostheses demonstrate equivalent durability, despite post-implantation tissue changes in the former material. PMID- 10461236 TI - Multiple valve replacement increases the risk of reoperation for structural degeneration of bioprostheses. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to analyze the results of reoperations for structural degeneration of bioprostheses, and to define a high risk population for reoperative procedures. METHODS: A series of 524 consecutive patients who had undergone a first reoperative replacement for a failed bioprosthesis between 1978 and 1998 was reviewed retrospectively. The reoperative procedure comprised 363 single valve replacements, and 161 multiple valve replacements. During the original procedure, 648 bioprostheses had been implanted in the mitral (n = 403), aortic (n = 220) and tricuspid (n = 25) positions. RESULTS: The mean interval between the original procedure and reoperation was 8.8 +/- 3.3 years. Tissue valve failure was revealed by recurrence of cardiac insufficiency in 70% of cases. The overall early mortality rate was 8%, but early mortality rates for elective single mitral and aortic reoperative valve replacements were only 3.9% and 4%, respectively. Early mortality following reoperation for single and multiple valve replacement was 6.0% and 12.4% respectively (p = 0.02). Other significant multivariable predictors for early mortality were old age (p = 0.003), NYHA functional class (p = 0.007), presence of ascites (p = 0.02) and reoperation performed before 1988 (p = 0.013). CONCLUSIONS: The risk of reoperation for structural degeneration of bioprostheses is acceptable for elective single reoperative valve replacement as opposed to multiple reoperative valve replacement. This may limit the use of bioprostheses during the original procedure when multiple valve replacement is required. PMID- 10461237 TI - Inter-laboratory comparisons: approaching a new standard for prosthetic heart valve testing in vitro. AB - Current standards governing the evaluation of prosthetic heart valve designs have come under scrutiny. Generally, standards require measurements of pressure drops and regurgitant flow. While this information is important in the characterization of valve performance, these standards are both insufficient and ambiguous. Their insufficiency is due to the fact that they do not cover issues related to thrombosis and structural damage, and their ambiguity is demonstrated by the fact that different pulse duplicators will produce different results for nominally the same set of conditions. While the insufficiency of the current standards has recently been addressed, the ambiguity has not been addressed in a systematic way except for one particular study involving two pulse duplicator systems. This paper explores physical sources for disagreement in pressure and flow measurements between pulse duplicators, and suggests ways to account for them. By considering these physical phenomena, standards can be developed for testing chambers that improve similarity between systems. This should not compromise innovation in the design of new pulse duplicators, which may be necessary to address additional concerns besides the pressure and flow characteristics of the valve. PMID- 10461238 TI - Perioperative bleeding and thromboembolic risk during non-cardiac surgery in patients with mechanical prosthetic heart valves: an institutional review. AB - BACKGROUND AND AIMS OF THE STUDY: The study aim was to determine the risk of thromboembolic and bleeding complications in patients with mechanical heart valve prostheses who underwent non-cardiac surgery under different regimens of perioperative anticoagulation. Data were analyzed on the basis of surgery type and underlying disease. METHODS: A series of 235 patients (mean age 63 +/- 4.5 years) with one or two mechanical heart valves underwent subsequent non-cardiac surgery comprising abdominal, vascular and thoracic, orthopedic, urologic, neurosurgery, ENT, plastic and reconstructive, and gynecologic operations. Mean interval between heart valve replacement and non-cardiac surgery was 3.9 +/- 3.3 years. Perioperative oral anticoagulation was managed by discontinuation of oral anticoagulation and intravenous heparin administration; or by discontinuation and early postoperative re-institution of oral anticoagulation without intravenous heparin; or by no withdrawal of oral anticoagulation. Patients with bioprostheses were excluded. RESULTS: Overall hospital mortality during non-cardiac surgery was 2.9%. Thromboembolic events included cerebral embolism with transient deficit (n = 3), residual defect (n = 1) and irreversible defect (n = 1), as well as peripheral embolism (n = 11). Hemorrhagic complications included wound hematoma (n = 10) and increased postoperative bleeding (n = 8) with re-exploration in five patients. Thromboembolic complications occurred most often in patients with prosthetic mitral valve and atrial fibrillation; the lowest risk was in patients with sinus rhythm after aortic valve replacement. Most complications occurred after discharge and in patients with surgery for malignancy, within 10 days of instituting oral anticoagulation, and despite a therapeutic INR value. CONCLUSIONS: Minor surgical procedures can be performed safely without discontinuing anticoagulation. When major non-cardiac surgery is planned, discontinuing oral anticoagulation and starting perioperative intravenous heparin minimizes bleeding and thromboembolic risks. Thromboembolic complications may occur within one month of surgery, despite adequate oral anticoagulation, though permanent morbidity is low. PMID- 10461239 TI - Correlation of valve closing to visualization of single-leg separation in radiographic images of Bjork-Shiley convexo-concave heart valves. AB - BACKGROUND AND AIM OF THE STUDY: The study goal was to determine whether the visualization of single-leg separation (SLS) in cineangiographic sequences of Bjork-Shiley convexo-concave heart valves could be correlated to the position of the occluder disk within the cardiac cycle. METHODS: Images from ten patient cases with SLS valves were reviewed by three experts, who identified the image frames within a cine sequence that appeared suspicious for SLS. The position of the occluder disk, the frame rate, and the length of the cardiac cycle were noted relative to these image frames. RESULTS: The probability of detecting a SLS was not significantly correlated to any of these factors. CONCLUSIONS: Visualization of SLS in cineangiographic images is limited to a few frames within an imaging sequence. It appears that other features within the image play a larger role in a clinician's ability to detect a fracture than do the cardiac dynamics of the system. PMID- 10461240 TI - Videoassisted thrombectomy of mechanical prosthetic heart valves. AB - BACKGROUND AND AIM OF THE STUDY: Thrombosis of a bileaflet mechanical heart valve is a life-threatening clinical event. Surgical thrombectomy of bileaflet mechanical prostheses remains an appropriate treatment in selected patients. METHODS: Between 1996 and 1998, five patients (three men, two women; average age 56 +/- 1 years; range: 56 to 66 years) with thrombosis of left-sided bileaflet mechanical valves were treated with videoassisted thrombectomy of the prosthesis. Four patients had thrombosis of a bileaflet mitral mechanical valve, and one patient had thrombosis of an aortic valve prosthesis. Preoperatively, patients were in either NYHA functional class IV (n = 4) or class I (n = 1). Surgery was performed through a right anterior thoracotomy or a median sternotomy. A rigid 30 degrees thoracoscope was inserted into the left atrium or aorta to visualize the thrombosed valve. The thrombus was extracted and the prosthesis under-surface examined and cleaned. Leaflet mobility, assessed with transesophageal echocardiography, was normal following surgical thrombectomy. RESULTS: Mean cardiopulmonary bypass time was 102 +/- 30 min; mean aortic cross-clamping time was 47 +/- 25 min. There was no hospital mortality; mean hospital stay was 9 +/- 1 days (range: 6 to 11 days). Anticoagulation with intravenous heparin was resumed 24 h after surgery. Three patients were discharged on coumarin treatment alone; two patients received aspirin plus coumarin. Mean postoperative follow up was 7 +/- 8 months (range: 1 to 21 months). One patient died 21 months after thrombectomy of a mitral prosthesis, with an unconfirmed diagnosis of recurrent mitral valve dysfunction. At 1-15 months after surgery, four patients are in NYHA class I, without evidence of prosthesis dysfunction. CONCLUSION: Videoassisted thrombectomy of a bileaflet mechanical heart valve is a treatment option in patients with acute thrombosis of the prosthesis. PMID- 10461241 TI - Aortic root geometry: pattern of differences between leaflets and sinuses of Valsalva. AB - BACKGROUND AND AIMS OF THE STUDY: Growing interest in aortic root replacement with the use of stentless auto-, homo- and xenografts, and new developments in aortic valve conservation demand a deeper understanding of the normal aortic root anatomy. METHODS: Ten cryopreserved human aortic roots were pressurized, fixed and measured directly (leaflet free edge and attachment) and using three dimensional computed tomography imaging software (sinus of Valsalva height and volume). RESULTS: The mean of the measurements of all four parameters yielded a pattern in which the non-coronary sinus (N) structures had the greatest dimensions followed by the right (R) and then the left (L). Non-parametric ANOVA on each of these parameters also showed significant differences among the sinuses yielding a pattern of N > R > L. This pattern determined an angle of tilt between the plane at the base (annulus) and the plane intersecting the sinotubular junction with a mean value of 11 degrees. Linear regression indicated that this angle did not depend on the size of the base (annulus). CONCLUSIONS: The data showed a geometric pattern of the aortic root, with the structures of the non coronary sinus being the largest followed by the right and then the left. The possible hemodynamic relevance and surgical implications of these findings need to be explored. PMID- 10461242 TI - Aortic stenosis: an atherosclerotic disease? AB - Aortic stenosis is a common disorder, which affects an increasing number of elderly patients, and is responsible for considerable morbidity. Although some associations for this disorder have been described, factors driving its progression remain obscure. This review proposes the hypothesis that aortic stenosis is another manifestation of atherosclerotic disease, and that its progression can be explained by classical cardiovascular risk factors. PMID- 10461243 TI - Surgical repair in ruptured congenital sinus of Valsalva aneurysms: a 13-year experience. AB - BACKGROUND AND AIM OF THE STUDY: Rupture of congenital sinus of Valsalva aneurysm is a rare cardiac malformation that usually causes reduced cardiac performance. METHODS: Twenty patients (mean age 28.3 +/- 10.7 years; range: 14 to 55 years) with rupture of congenital sinus of Valsalva aneurysm were operated on at our institution between January 1985 and March 1999. The origin of the ruptured aneurysms was the right coronary sinus in 18 patients (90%) and the non-coronary sinus in two (10%). No ruptures were observed originating from the left coronary sinus. The aneurysms ruptured into the right ventricle in 14 patients (70%), into the right atrium in five (25%), and into the left ventricle in one patient (5%). Subarterial ventricular septal defect (VSD) was the most common associated defect (30%), and aortic insufficiency the second (20%). No correlation was found between subarterial VSDs and aortic insufficiency (p > 0.05). To achieve repair, the aorta and cavity into which the aneurysm had ruptured were opened. The aneurysmal sac was excised and the defect closed with a patch in 18 patients, and without patch in two. RESULTS: One patient died in hospital (mortality rate 5%); no late mortality was observed. Surviving patients were followed up for 4.4 +/- 3.3 years (range: 1 to 13 years). The actuarial survival rate was 95% at 12.35 years. Recurrence of fistula was observed in one patient (5%) in whom the ruptured aneurysm had been closed by primary suture without the use of a patch. At 12.35 years, the actuarial freedom from recurrence of fistula was 94.74% for all survivors, and 100% for patients with patch closure. No late complications have been observed. CONCLUSIONS: Following diagnosis of ruptured sinus of Valsalva aneurysm, surgical repair is the treatment of choice. Surgery performed as rapidly as possible after diagnosis in general leads to an excellent outcome. PMID- 10461244 TI - Exercise-induced hyperkalemia and concentration of Na,K-pumps in skeletal muscle in mitral stenosis: effect of balloon mitral valvotomy. AB - BACKGROUND AND AIMS OF THE STUDY: The study aim was to examine the effects of balloon mitral valvotomy (BMV) on exercise-induced hyperkalemia, and on changes in the concentration of Na,K-pumps in skeletal muscle, as an exaggerated exercise induced rise in potassium concentration ([K+]) may contribute to exertional fatigue and breathlessness. METHODS: Eight subjects were evaluated with mitral stenosis (mean age 34 +/- 5.2 years) before, and at two weeks and four months after BMV. Subjects underwent incremental exercise to exhaustion for exercise induced rise in [K+] and vastus lateralis biopsy for concentration of Na,K-pumps. RESULTS: Mean (+/- SE) valve area increased from 0.89 +/- 0.03 cm2 before to 1.75 +/- 0.05 cm2 after BMV. There was a progressive increase in VO2,max (15.3 +/- 1.6, 17.2 +/- 1.4 and 19.9 +/- 1.9 l/kg/min) at baseline, early after and later after BMV, respectively (p < 0.01). The rise in [K+] with absolute workload fell progressively at early and late follow up post-BMV (p < 0.05), but was unchanged when plotted against percentage of VO2,max to match for relative workload. The concentration of Na,K-pumps was similar to baseline at early follow up (233 +/- 10 versus 228 +/- 15 pmol/g wet weight), but was significantly increased at late follow up after four months (265 +/- 17 pmol/g; p < 0.05). When the relationship between the concentration of Na,K-pumps and the exercise-induced rise in [K+] was studied, a negative correlation was found. However, correlation analysis for the effects of changes in Na,K-pumps on changes in exercise hyperkalemia after BMV was not significant. CONCLUSIONS: The progressive reduction in exercise-induced rise in [K+] after BMV may contribute to the progressive improvement in exercise performance. The increased concentration of Na,K-pumps in skeletal muscle may assist in this improvement, and emphasizes the importance of peripheral adaptations in clinical improvement after BMV. PMID- 10461245 TI - Extended vertical transseptal approach versus conventional left atriotomy for mitral valve surgery. AB - BACKGROUND AND AIM OF THE STUDY: Mitral valve surgery requires optimal exposure of the valvular apparatus, particularly when a conservative procedure is used. Retrospectively, we compared surgical results in patients who underwent mitral valve surgery using the vertical transseptal approach (which has been adopted routinely in our institute) with those in patients undergoing conventional left atriotomy. METHODS: A total of 172 consecutive patients operated on for mitral procedures were allocated to either group A (those operated on through a longitudinal left atriotomy; n = 62), or group B (mitral valve exposure achieved through an extensive vertical transseptal approach; n = 110). RESULTS: In group A, there were 24 valvular reconstructions and 38 valvular replacements. Mean (+/- SD) cardiopulmonary bypass (CPB) time was 65.9 +/- 17 min and mean ischemia time 37.4 +/- 13 min. Total postoperative bleeding was 277 +/- 171 ml. There was no surgical re-exploration for bleeding. One patient in this group died (mortality rate 2%). Among 40 patients in atrial fibrillation preoperatively, four had one episode of temporary junctional rhythm, six had temporary sinus rhythm and two had stable sinus rhythm. Among patients with preoperative sinus rhythm, six (27%) had episodes of atrial fibrillation and two (9%) temporary atrioventricular block. In group B, 46 mitral reconstructions and 64 mitral replacements were performed. Mean CPB time was 67.9 +/- 20 min and mean ischemia time 48.1 +/- 17 min. Total postoperative bleeding was 400 +/- 189 ml. Three patients in this group died (mortality rate 2.7%). Among 60 patients with preoperative atrial fibrillation, six (10%) had one episode of temporary junctional rhythm, 14 (24%) had temporary sinus rhythm and two (3%) had conversion to stable sinus rhythm. Among those in sinus rhythm preoperatively, 16 (32%) had episodes of temporary junctional rhythm, two (4%) had temporary atrial fibrillations, and four (8%) had stable atrial fibrillation. In group B patients, the incidences of ischemia time and total postoperative bleeding (p = 0.004), and postoperative junctional arrhythmia in those with preoperative sinus rhythm (p < 0.001), were greater than in group A patients. CONCLUSIONS: No technique-related deaths occurred; neither were causes of re-exploration for bleeding related to technique, and there was no evidence of residual interatrial shunt. In conclusion, transseptal extended atriotomy provides excellent exposure for mitral valve surgery. Disadvantages of minimally increased ischemia time and surgical bleeding are minor compared with the superior and more complete surgical reconstruction achieved. The advantages of the technique are undermined by the higher incidence of junctional arrhythmia which, even if temporary, requires strict postoperative monitoring. PMID- 10461246 TI - Aortic stenosis in endogenous ochronosis. AB - Endogenous ochronosis, a rare inherited disease of tyrosine metabolism, is caused by a deficiency of the enzyme, homogentisic acid oxidase, and may lead to cardiovascular involvement seen most frequently as aortic valve stenosis. We report the case of a patient with generalized ochronosis who developed cardiovascular symptoms due to aortic valve stenosis and who underwent aortic valve replacement. PMID- 10461247 TI - Combined mitral and tricuspid valve repair in acute infective endocarditis. AB - Combined repair of the mitral and tricuspid valves involved with acute infective endocarditis was carried out in a 38-year-old drug addict. Mitral valve repair included vegetectomy, closure of posterior leaflet perforation, and posterior annuloplasty with a patch and a strip of glutaraldehyde-tanned autologous pericardium, respectively, while the tricuspid valve was reconstructed with the use of artificial chordae and valve bicuspidalization. At five months follow up the patient is asymptomatic, with echocardiographic evidence of only trivial mitral and tricuspid incompetence, and no signs of recurrent infection. This case report supports the use of valve reconstruction as a valuable option in patients in whom there is simultaneous involvement of the mitral and tricuspid valves with infective endocarditis. PMID- 10461248 TI - Asymptomatic left ventricular outflow tract obstruction after mitral valve replacement with leaflet preservation. AB - It has been shown that inappropriate anterior mitral leaflet preservation during mitral valve replacement may cause left ventricular outflow tract (LVOT) obstruction, usually with dismal prognosis. In this report, we describe a patient with chronic asymptomatic LVOT obstruction after mitral valve replacement with leaflet preservation. PMID- 10461249 TI - Successful Cox-maze III procedure combined with mitral valve replacement in a massively thrombosed left atrium. AB - A case is reported of chronic atrial fibrillation resulting from long-standing rheumatic mitral stenosis complicated by a massively thrombosed left atrium. In this patient, restoration of sinus rhythm and atrial transport function was performed using a Cox-maze III procedure with mitral valve replacement. PMID- 10461250 TI - Isolated tricuspid valve regurgitation resulting from severe annular dilatation: case report. AB - A rare case of isolated tricuspid regurgitation (TR) in a 65-year-old man is presented. Echocardiography revealed enlargement of the right atrium, dilatation of the tricuspid valve annulus without thickening or prolapse of the leaflets, and an intact atrial septum. No downward displacement of the tricuspid septal leaflet was observed by echocardiography. Mild mitral regurgitation and severe TR were detected on color flow Doppler studies. Cardiac catheterization indicated elevated right atrial pressure, with a pronounced V-wave. No left-to-right shunt was detected at the right atrium. At surgery, severe annular dilatation of the tricuspid valve (without organically diseased or deformed tricuspid leaflets) was observed, and tricuspid annuloplasty with a prosthetic ring performed. Postoperative echocardiography and right ventriculography showed trivial TR. PMID- 10461251 TI - Conservative surgery for atrioventricular valve myxoma. AB - Patients with valvular myxoma are usually candidates for surgery because of the high incidence of life-threatening embolism. In some cases, the tumor is sessile or presents with a large peduncle: complete excision may then lead to valve replacement. We report two cases of atrioventricular valve myxoma where replacement was avoided. In one patient, a mitral myxoma appended from the edge of the anterior leaflet close to the chordae insertion; safe excision implied destruction of the two chordae and a peritumoral section of the anterior leaflet. A chordal transposition technique was used to preserve valve competence. In a second patient, a tricuspid myxoma causing syncopal episodes was resected; this was characterized by a large stalk, located on the anterior tricuspid leaflet away from chordal attachment and the valvular annulus. Treatment was by resection and the leaflet reconstructed with a pericardial patch. Techniques for conservative treatment of degenerative valvular disease or endocarditis, when monitored peroperatively by transesophageal echocardiography, may be successful in the surgical resection of atrioventricular myxoma. PMID- 10461252 TI - Ehlers-Danlos syndrome: surgical management of mitral regurgitation and atrial fibrillation. AB - Ehlers-Danlos syndrome is associated with fatal cardiovascular complications and intraoperative death. The lack of literature on cardiac surgery in the syndrome makes it difficult for surgeons to assess the risk of an operation and to choose optimal treatment. We describe the case of a 63-year-old man with Ehlers-Danlos syndrome type I or II, mitral regurgitation and atrial fibrillation who was managed successfully with mitral valve repair and surgical cardioversion. PMID- 10461253 TI - Screening of high-risk infants using distortion product otoacoustic emissions. AB - Otoacoustic emissions have been used as an objective and noninvasive test of cochlear function. Due to its ease of use, accuracy, and test-retest reliability, otoacoustic emissions testing is considered an excellent tool for evaluating difficult-to-test patients. This project involved screening infants at high risk of congenital hearing loss. These infants were selected from the Special Infant Clinic at Queen's University, Kingston, Ontario. The first part of this project was to establish a distortion product otoacoustic emissions (DPOAEs) "norm" as the reference for this group of patients. A prospective study was then conducted to compare DPOAE and auditory brainstem response (ABR) as screening tools for this group of patients. Auditory brainstem response has been the gold standard for evaluating the hearing status of these infants. We will also present our experience in testing over 50 patients at high risk of congenital hearing loss using DPOAEs. PMID- 10461254 TI - Evaluation of immunotherapy by nasal antigen challenge. AB - BACKGROUND: Nasal antigen challenge test is used to evaluate the allergic conditions within different diagnostic procedures. OBJECTIVE: This study was conducted to evaluate the efficacy of immunotherapy (IT) in an objective and simple way in children with perennial allergic rhinitis. METHOD: This study was a double-blind, placebo-controlled procedure. Nineteen patients (mean age 10 +/- 6 years) were treated only with IT and 17 patients (mean age 11 +/- 5 years) constituted the control group without any treatment. The patients were evaluated at the beginning and at 3 and 6 months of IT. Nasal provocation reactions were evaluated and scored at 5-, 10-, and 30-minute intervals. RESULTS: In the IT group, mean clinical scores and mean nasal provocation scores decreased significantly between the initial value and at 3 and 6 months of IT, whereas there was no significant change in the value for the control group. There were no significant differences among the 5-, 10-, 15-, and 30-minute evaluations within both the IT and control groups. CONCLUSION: The efficacy of IT in perennial allergic rhinitis may be assessed by the nasal antigen challenge test in a simple and objective way within a 5-minute period. PMID- 10461255 TI - Scintigraphic assessment of swallowing after horizontal supraglottic laryngectomy. AB - OBJECTIVE: Oral-pharyngo-esophageal scintigraphy (OPES) proved to be able to demonstrate the pharyngeal residue of the radioactive bolus after deglutition and the presence and amount of tracheobronchial aspiration, and to calculate exactly the transit time of the various swallowing phases. The aim of this study was to evaluate the real objective effectiveness of OPES in order to assess the degree of swallowing recovery after horizontal supraglottic laryngectomy (HSL). METHODS: Nineteen patients entered the study, and 17 healthy subjects were included as control group (group 1). Nineteen patients in whom HSL had been performed at least 1 year before underwent OPES. They were divided into two groups: 13 patients (group 2) in whom the tracheal cannula had been removed and swallowing, phonatory, and respiratory functions were satisfactory and 6 patients (group 3) in whom the tracheostomy tube was still in situ for aspiration of liquids and scarring of the laryngeal vestibule. RESULTS: Our results showed that in the patients who underwent HSL, all scintigraphic semiquantitative parameters and particularly aspiration percentage values and Pharyngeal Ritention Index (PRI) at 10 and 60 seconds were able to pinpoint some residual "subclinical" alteration and/or minimal surgical sequelae frequently observed after this kind of functional surgery, even though a substantially satisfactory recovery was achieved clinically (group 2 versus group 1). In addition, aspiration amounts in group 3 were markedly higher than those in group 2 (p < .0002), and mean PRI values at 10 and 60 seconds were significantly different (p < .0001) from normal mean control values (group 1). CONCLUSION: Oral-pharyngo-esophageal scintigraphy may be regarded as a noninvasive, well-tolerated technique, with a radiation body burden within satisfactory limits of radioprotection. It appeared to be also sensitive in assessing the swallowing recovery after HSL. PMID- 10461257 TI - Vestibular rehabilitation on a budget. AB - OBJECTIVE: This study was conducted to compare the results of an office-based vestibular rehabilitation program with the results from other larger programs in the literature. METHOD: Using a retrospective chart review, 138 dizzy patients seen in a tertiary medical centre were asked to perform directed exercises. Significant subjective improvement reported by the patients. RESULTS: Sixty-six percent of patients improved with this form of vestibular rehabilitation after 1 month. The results compare favourably with improvement rates from other centres in the literature with large, multidisciplinary teams. CONCLUSION: Vestibular rehabilitation therapy can be effectively administered from an office-based program. PMID- 10461256 TI - Does nasal nitric oxide come from the sinuses? AB - OBJECTIVE: The purpose of this study was to assess nitric oxide (NO) output by the nose and sinuses. METHOD: In one volunteer, the osteomeatal complex and sphenoethmoidal recess were occluded to isolate the nose from the sinuses. The antrum and frontal sinus were each punctured by two catheters and irrigated with air at constant flow. Nitric oxide output and its rate of accumulation in the absence of air flow were measured in each sinus and in the adjacent nasal cavity. RESULTS: Prior to ostial occlusion, NO output in the nose was 96 nL/min. It decreased by 12% after blockage of all of the ostia. In the isolated sinuses, it was 190 nL/min (antrum) and 68 nL/min (frontal). After 5 minutes stagnation; NO concentration [NO] rose in the occluded sinuses to 24,700 nL/L in the antrum and 22,300 nL/L in the frontal sinus. In the nose, it increased to 29,000 nL/L. When the period of stagnation was prolonged in the frontal sinus, the [NO] reached a plateau. NO output and accumulation were not altered in the nose or either sinus by opening their ostia. In the antrum and frontal sinus, lidocaine reduced NO output and the rate of NO accumulation, but not in the nose. CONCLUSIONS: In this volunteer, 88% of nasal NO was derived from the nose itself. Nitric oxide exchange between the frontal sinus, antrum, and nose was negligible. In the absence of air flow, [NO] rose to a plateau in the nose and frontal sinus. Lidocaine inhibited NO output in the sinuses but not the nose. PMID- 10461258 TI - Argyremia in septal cauterization with silver nitrate. AB - OBJECTIVE: The purpose of this study was to demonstrate silver absorption in blood and hair specimens after septal cauterization with silver nitrate and to discuss the potential toxicity of silver. METHOD: A prospective study of 11 volunteers without any known occupational exposure to silver products or past history of septal cauterization with silver nitrate was undertaken. Subjects were recruited in an academic tertiary care centre from October 1996 to September 1997. The study population consisted of five patients with anterior epistaxis and six healthy volunteers without any bleeding problem. Cauterization was done with one or two silver nitrate applicators directly on the bleeding vessel or Kiesselbach's area. Blood was sampled before cauterization and at specified times after application, while hair strands were sampled only 3 months later. Measurements of silver concentration in whole blood and in hair segments were obtained by inductively coupled plasma mass spectrometry. RESULTS: Silver concentrations in whole blood increased significantly after cauterization (p = .02). The measured peak level seemed to correlate with the number of applicators used. No significant increase in silver concentration was observed in hair samples. CONCLUSIONS: Effective silver absorption occurs with only one or two silver nitrate applicators. Hair has not been as reliable as whole blood to document an acute and fragmentary exposure. The indiscriminate use of silver nitrate is a potential source of silver intoxication. PMID- 10461259 TI - Surgical anatomy of the ear of the fat sand rat. AB - OBJECTIVE: This study was conducted to investigate and describe the anatomical details of the ear of the fat sand rat (Psammomys obesus). METHODS: Thirty ears (15 in dry skull and 15 in live animals) were dissected with the aid of an operating microscope and microsurgical techniques. Photographs were taken through an operating microscope. RESULTS: The temporal bone of the fat sand rat consists mainly of an unusually large bulla. Three distinct auditory ossicles were visualized and the manubrium of the malleus is relatively long in relation to the size of the ear. Most parts of the inner ear bulge into the bulla cavity and are easily accessible. The cochlea consists of 3.25 turns. The tympanic membrane, facial nerve, stapedial artery, and eustachian tube are described in detail. The large size of the bulla is probably related to the high sensitivity, particularly at low frequencies of the animals ear. CONCLUSION: The fat sand rat was found to be an excellent experimental animal for the investigation of middle and inner ear physiology. PMID- 10461260 TI - Management of congenital subglottic hemangioma. AB - OBJECTIVE: This study was conducted to assess the safety and efficacy of individualized management of congenital subglottic hemangioma (CSH) at the Children's Memorial Hospital in Chicago; describe treatment modalities, including endoscopic excision with the carbon dioxide (CO2) laser, systemic corticosteroids, and intralesional corticosteroid injection with short-term intubation; and determine the success of these various strategies in avoiding tracheotomy. METHODS: During the 10-year period between January 1, 1988 and December 31, 1997, 28 infants were diagnosed with CSH. A retrospective review of medical records was undertaken to determine demographics, presenting symptoms, location of the lesion, therapeutic modality, and complications. All patients were contacted at the time of writing. RESULTS: Twenty-eight patients met the criteria for this study. Gender distribution was 1.8:1 female to male. Age at diagnosis ranged from 4 weeks to 8 months, with a mean of 78.8 days and a median of 60 days. The location of CSH was most often posterior and on the left. Associated hemangiomas were found in 14 (50%) cases. The most common symptoms were stridor and cough. Management included 1 to 13 operative direct laryngoscopies and bronchoscopies, endoscopic excision with the (CO2) laser, and the use of systemic and intralesional corticosteroids. No patient required tracheotomy. CONCLUSION: Morbidity and the need for tracheotomy in CSH patients can be minimized using a combination of therapeutic modalities. Each infant's treatment is individualized based on the severity of the patient's symptoms and the morphology of the lesion. PMID- 10461261 TI - Laryngeal scleroma. AB - Respiratory scleroma is a chronic, progressive granulomatous disease of the respiratory tract. The causal organism is the Klebsiella rhinoscleromatis. The disease has three stages: the initial catarrhal stage, granulomatous stage, and sclerotic stage. The object of this report was to determine the clinical behaviour of the laryngeal scleroma. The study included 17 patients with this manifestation characterized by airway obstruction and dysphonia. The report revealed that respiratory scleroma affected the larynx in 40%, and the principal findings were glottic/subglottic stenosis. PMID- 10461262 TI - Measurement of nasal geometry by acoustic rhinometry in normal-breathing Asian children. AB - OBJECTIVE: The purpose of this study was to obtain normal values of nasal geometry in Asian children with no nasal problems using the acoustic rhinometry method. METHOD: Acoustic rhinometric measurement of the nasal fossae was performed in pediatric patients coming for general surgical operations with no nasal problem. RESULTS: A cohort study of 183 children in an acute care hospital with full elective and emergency surgical services was undertaken. The mean minimal cross-sectional area (MCA), which was functionally important for nasal breathing, was found to be 0.32 cm2 (SD = 0.13) and situated at 1.40 cm (SD = 0.26) from the anterior nare. Normal values for area, length, and volume of the nose in these children as a group and at different ages from 1 to 11 years old were obtained. CONCLUSIONS: Acoustic rhinometry was found to be well tolerated by the children. The MCA was confirmed to be situated at the anterior part of the nose and had a positive correlation with an increase in age. PMID- 10461263 TI - Evaluation system for facial skin assessment. PMID- 10461264 TI - Paranasal sinus scleroma. PMID- 10461265 TI - A terminally ill patient wants to die--can you be the judge? PMID- 10461266 TI - Surgery to heal the psyche? PMID- 10461268 TI - Personality testing--the short form. PMID- 10461267 TI - Anxiety plays a major role in persistent depression. PMID- 10461269 TI - The 'symptom' of alternative therapy in breast cancer. PMID- 10461270 TI - Treating pediatric anxiety may prevent adult disorders. PMID- 10461271 TI - Healthy people 2000--how are we doing? PMID- 10461272 TI - Well siblings living with schizophrenia. Impact of associated behaviors. AB - As a result of the ongoing hallucinations and delusions, life at home became "anxious and chaotic" with "lots of stress." Because there was no on-site crisis intervention by professionals, well siblings were sometimes called upon to control the violent behavior and were vigilant and fearful regarding the potential for abuse. Negative symptoms were the most disturbing to well siblings. Siblings need help to understand that social isolation and lack of motivation are symptoms of the illness--not due to "laziness." PMID- 10461273 TI - Spousal caregiving when the adult day services center is closed. AB - Of those spousal caregivers who identified a time when they most needed assistance, morning or afternoon time periods appeared to be most problematic. Spousal caregivers who reported receiving respite time on the weekends identified adult children as the informal support person most often available. However, more than half of the caregivers reported having no respite time during the weekend. Differences in the ways spousal caregivers spent their respite time on the weekends, if available, varied by gender, length of tenure as a caregiver, and age. PMID- 10461274 TI - Toward a cultural conception of the self. AB - The dominant mental health models rely excessively on a modern Western ideal of the self. Health services in a pluralistic society require interventional approaches that recognize the significance of culture in shaping a person's conception of self as well as mental health and illness. PMID- 10461275 TI - Images & relationships forged in war. A study of women nurses who served in Vietnam. AB - Remembrance of visual images and other sensations of war lasts a long time after the war is over, and these memories may be painful and disruptive for years. Training of military nurses and other health care personnel for war or disaster response should emphasize lessons learned from past wars and disasters. Curricula to meet the clinical and psychological challenges of war and disaster nursing will better prepare those who serve, and will lead to a more realistic understanding of situational demands and expectations. PMID- 10461276 TI - Who was John Howard? AB - John Howard was an 18th-century English philanthropist who made significant contributions in prison reform. Despite personal tragedy and an oppositional social climate, he became an early promoter of humane treatment for prisoners. Other reformers followed John Howard, making valuable contributions, but many challenges remain in the management of forensic hospitals and prison systems. Howard's legacy is not only the modernization of prison structures and programs, but also the work of numerous worldwide societies and associations that provide services for communities and prisoners. PMID- 10461277 TI - Asset building: rhetoric versus reality--a cautionary note. PMID- 10461278 TI - Building developmental assets: response to Price and Drake. PMID- 10461279 TI - "Ouch, my aching back, again!". PMID- 10461280 TI - Encouraging schools to promote health: impact of the Western Australian School Health Project (1992-1995). AB - The Western Australian School Health (WASH) Project, a school health promotion intervention operating over a four-year period (1992-1995), provided a comprehensive, year-long intervention to help successive groups of schools develop health promotion programs. The WASH Project worked with self-selected school communities and used community development strategies to support participating schools in identifying and responding to health concerns relevant to their students. This paper reports the school impact results of the WASH Project. School impact data involved 24 variables categorized into two areas: school organizational factors supportive of health promotion, and school health promotion factors. Two methods of analysis were used: logistic regression indicating the direction of change, and linear regression indicating the magnitude of change. Results demonstrated that schools successfully made organizational changes, such as the allocation of additional time, personnel, and monetary resources, to support health promotion. PMID- 10461281 TI - Lessons learned from the Hunter Region Health Promoting Schools Project in New South Wales, Australia. AB - This paper describes development, over a five-year period, of the Hunter Region Health Promoting Schools Project in New South Wales (NSW), Australia. The project implemented the principles and philosophy of the Health Promoting Schools concept and evaluated its effectiveness using a randomized controlled trial involving 22 public secondary schools. An overview of the preliminary intervention model based on the health promoting schools philosophy and trialed in a pilot study is provided. The authors also outline barriers to, and difficulties in, implementing the philosophically based intervention model in the secondary school setting. The current intervention approach, which evolved over five years, is described in relation to the roles played by project team members and school communities. In addition, the authors outline guiding principles arising from the new approach which facilitated adoption of health promotion strategies in secondary school settings. PMID- 10461282 TI - Asthma knowledge, roles, functions, and educational needs of school nurses. AB - In October 1997, 790 school nurses in Maryland and the District of Columbia were surveyed to determine their attitudes, knowledge, and beliefs about asthma. Results for 550 (70%) nurses indicated school nurses possess a generally appropriate level of knowledge concerning asthma, and most asthma myths have been replaced with knowledge. However, school nurses also have varied responsibilities that affect their ability to provide health education and support services to children with asthma at school. Little time is available for a proactive role. Concerns about the criteria and follow-up for delegating medication administration within the school setting were reported. A lack of communication existed with parents about the child's asthma. Open communication between school nurses and the family is recommended to establish a partnership and improve asthma management outcomes. In addition, school policies and procedures should be updated to meet the demands of children with asthma. PMID- 10461283 TI - The status of middle level HIV/STD education as assessed by state and local education agencies. AB - During spring 1998 RAD Educational Programs, in cooperation with the National Middle School Association, conducted a needs assessment of middle level HIV/STD prevention programs from the perspective of representatives from state education agencies (SEA) and local education agencies (LEA). Data were collected from 84% of states (SEAs) and 88% of cities (LEAs) funded by CDC to conduct HIV/STD prevention activities. A range of activities are occurring in school-based HIV/STD prevention for early adolescents in grades 5-8. However, a prevailing characteristic among states was the high degree to which local control and local decision making affects programming in schools. Needs of SEAs and LEAs are similar as reflected by the need for continued improvement of middle level HIV/STD prevention education. A concrete result was the multitude of requests for effective HIV/STD curricula for middle level students. In addition, administrators shared similar needs for an increase in time allotment for health instruction. Most SEAs (65%) and LEAs (77%) believed that time for health education in the classroom would increase if a nation-wide mandate existed resulting in appropriate standards and assessments for all middle level and high school students. PMID- 10461284 TI - Ontario, Canada, high school teachers as enforcers in the war on drugs: what their students see and say. AB - This paper examines how high school teachers interact with students on the subject of marijuana. Results, based on 49 focus groups with 278 high school students in Ontario, Canada, reveal three basic assumptions of the students: 1) only some teachers can actually tell when a student is high on marijuana; 2) many teachers have tried marijuana or continue to use it; and 3) individual teachers vary in how they respond to students who are high. Results suggest that changes in the relationship between marijuana and authority account in large part for the seeming reluctance of so many teachers to exercise their mandate to discipline students who use marijuana. The reasons for this are twofold: 1) many teachers do not see use of marijuana of and by itself as a threat to their authority; and 2) teachers who choose to confront users run the risk of having their authority and independence of action undermined once they report infractions to administrators who have authority over teachers and students alike. PMID- 10461285 TI - Electronic portfolios enhance health instruction. PMID- 10461286 TI - CABG and ICD for all patients with hemodynamically significant ventricular arrhythmia and significant coronary artery disease? Do we know enough to decide- or to design a randomized trial? PMID- 10461287 TI - Coronary artery bypass grafting and defibrillator implantation in patients with ventricular tachyarrhythmias and ischemic heart disease. AB - In patients with sustained ventricular tachyarrhythmias and myocardial ischemia due to multivessel coronary artery disease, it remains unclear whether revascularization is enough to control the arrhythmias or whether additional implantation of a defibrillator is indicated. We therefore reviewed our clinical strategy of performing both bypass surgery and implantation of a defibrillator in patients with syncopal ventricular tachycardia or fibrillation and significant multivessel coronary artery disease. We retrospectively reviewed the outcome of 18 patients with malignant ventricular tachyarrhythmias, significant multivessel coronary artery disease, and signs of myocardial ischemia who underwent both bypass surgery and defibrillator implantation. Data on these patients were compared to data from 232 other defibrillator patients with respect to baseline clinical variables, cardiac events, and mortality during follow-up. Except for underlying pathology, no other important differences in baseline characteristics were noted between the study patients and the other defibrillator patients. The cumulative occurrence of shocks during follow-up was comparable in both groups (66% vs 67%). The cumulative survival from all-cause mortality was 94% in the study patients and 78% in the others (P = NS). Pre- and postoperative electrophysiological testing was not useful to predict arrhythmia recurrences. In this population of patients with ventricular tachyarrhythmias and ischemia due to multivessel coronary artery disease, bypass surgery alone would not have prevented recurrences of arrhythmias. An excellent survival and a high incidence of shocks after both bypass surgery and defibrillator implantation were observed. PMID- 10461288 TI - A prospective, randomized, comparison in patients between a pectoral unipolar defibrillation system and that using an additional inferior vena cava electrode. AB - The decrease of defibrillation energy requirement would render the currently available transvenous defibrillator more effective and favor the device miniaturization process and the increase of longevity. The unipolar defibrillation systems using a single RV electrode and the pectoral pulse generator titanium shell (CAN) proved to be very efficient. The addition of a third defibrillating electrode in the coronary sinus did not prove to offer advantages and in the superior vena cava showed only a slight reduction of the defibrillation threshold (DFT). The purpose of this study was to determine whether the defibrillation efficacy of the single lead unipolar transvenous system could be improved by adding an electrode in the inferior vena cava (IVC). In 17 patients, we prospectively and randomly compared the DFT obtained with a single lead unipolar system with the DFT obtained using an additional of an IVC lead. The RV electrode, Medtronic 6936, was used as anode (first phase of biphasic) in both configurations. A 108 cm2 surface CAN, Medtronic 7219/7220 C, was inserted in a left submuscular infraclavicular pocket and used as cathode, alone or in combination with IVC, Medtronic 6933. The superior edge of the IVC coil was positioned 2-3 cm below the right atrium-IVC junction. Thus, using biphasic 65% tilt pulses generated by a 120 microF external defibrillator, Medtronic D.I.S.D. 5358 CL, the RV-CAN DFT was compared with that obtained with the RV-CAN plus IVC configuration. Mean energy DFTs were 7.8 +/- 3.6 and 4.8 +/- 1.7 J (P < 0.0001) and mean impedance 65.8 +/- 13 O and 43.1 +/- 5.5 O (P < 0.0001) with the RV-CAN and the IVC configuration, respectively. The addition of IVC significantly reduces the DFT of a single lead active CAN pectoral pulse generator. The clinical use of this biphasic and dual pathway configuration may be considered in patients not meeting implant criteria with the single lead or the dual lead RV-superior vena cava systems. This configuration may also prove helpful in the use of very small, low output ICDs, where the clinical impact of ICD generator size, longevity, and related cost may offset the problems of dual lead systems. PMID- 10461289 TI - Correlation waveform analysis to discriminate monomorphic ventricular tachycardia from sinus rhythm using stored electrograms from implantable defibrillators. AB - In order to examine whether a template-matching program utilizing correlation waveform analysis (CWA) might be used to discriminate monomorphic ventricular tachycardia (MMVT) from sinus rhythm (SR) in patients with implantable cardioverter defibrillators (ICDs), we studied stored episodes of induced MMVT in 25 patients and compared them to corresponding stored SR electrograms. We calculated mean correlation coefficients for SR beats against an SR template chosen within each sinus episode, induced MMVT beats against an induced MMVT template within each ventricular tachycardia episode, and induced MMVT beats against the original SR template. For each patient, the 99.5% lower confidence limit for the mean correlation coefficient of SR beats versus an SR template (patient-specific method) or the empirical correlation coefficient value 0.9 were selected as threshold values to discriminate induced MMVT from SR. The mean correlation coefficient for induced MMVT beats versus the original SR template for each patient was subtracted from both threshold values. A positive value is defined as accurate discrimination of induced MMVT from SR. Using 0.9 for a threshold cut off, 21 of 25 episodes of induced MMVT were accurately labeled with a sensitivity of 84%. Using the patient-specific method, we were able to correctly distinguish 23 of 25 episodes of induced MMVT from SR with a sensitivity of 92%. There was no statistically significant difference between the patient-specific or empirical methods in detecting MMVT (P 50.4). This is the first demonstration using stored intracardiac electrograms from ICDs that CWA is able to discriminate MMVT from SR with high sensitivity. Such a template-matching system may be used for off-line analysis or real-time rhythm discrimination. PMID- 10461290 TI - Experience with a low profile bipolar, active fixation pacing lead in pediatric patients. AB - Continued miniaturization of permanent pacing systems has promoted use of this technology in younger and smaller pediatric patients. Intermedics ThinLine 438-10 active fixation pacing leads (4.5 Fr lead body) were implanted in 26 patients (17 males/9 females; 9.9 +/- 6.9 years). Twenty of 26 patients received dual chamber systems, 6 of 26 patients single lead systems. Each patient has been followed 3 months. Pacemaker analysis at implant and 6 months later evaluated pulse width thresholds at 2.5 V (atrial 0.07 +/- 0.02 vs 0.13 +/- 0.02 ms [P = 0.01]; ventricular 0.08 +/- 0.04 ms vs 0.20 +/- 0.04 ms [P = 0.01]); sensing thresholds (atrial 4.1 +/- 0.41 mV vs 4.0 +/- 4.2 mV [P = NS]; ventricular 9.7 +/- 0.72 vs 9.3 +/- 0.94 mV [P = NS]); and impedance (atrial 345 +/- 12 vs 370 +/- 120 O [P = 0.04]; ventricular 412 +/- 17 vs 458 +/- 190 O [P < 0.01]). One volt lead failed with exit block at approximately 6 weeks. The youngest (9 months to 5 years) and smallest (6.5-18.0 kg) ten patients have each shown by venography to have at least mild venous stenosis at the lead(s) insertion site; five patients demonstrated collateral formation around asymptomatic obstruction, with no thrombus formation. The Intermedics 438-10 ThinLine pacing lead has demonstrated good and stable early postimplant electrical parameters. Angiographic evaluation in our smaller patients has shown evidence for asymptomatic venous obstruction. PMID- 10461291 TI - Evaluation of autonomic influences on QT dispersion using the head-up tilt test in healthy subjects. AB - Our objective was to examine the autonomic influence on QT interval dispersion using the head-up tilt test in healthy subjects. RR and QT intervals, heart rate variability, and plasma norepinephrine concentration were measured in the supine position and tilting to 70 degrees for 20 minutes using a footboard support in 15 healthy male volunteers (mean age +/- SD: 28.0 +/- 4.5 years). The rate-corrected QT interval (QTc) was calculated using Bazett's formula, and QT and QTc dispersions were defined as the maximum minus minimum values for the QT and QTc, respectively, from the 12-lead ECG. Spectral analysis of the heart rate variability generated values for the low- and high-frequency powers (LF and HF) and their ratio (LF/HF). Compared with values obtained in the supine position, tilting significantly increased QT (P < 0.05) and QTc dispersion (P < 0.01), the LF/HF ratio (P < 0.0001), and plasma norepinephrine concentration (P < 0.0001), and significantly decreased HF (P < 0.0001). QTc dispersion was positively correlated with the LF/HF ratio and plasma norepinephrine concentration, and negatively correlated with HF. These results suggest that head-up tilt testing increases QT dispersion by increasing sympathetic tone and/or decreasing vagal tone in healthy subjects. PMID- 10461292 TI - Time trends in the intracardiac potential recorded by pacemaker telemetry: comparison between steroid-eluting small area electrodes. AB - We assessed the time course of electrograms sensed both in the atrium and ventricle by two different steroid-eluting electrodes: Medtronic Capsure SP (with an area of 5.5 mm2) and Z (with an area of 1.2 mm2). We considered 68 unipolar electrodes: 31 atrial (19 Capsure SP 4523 and 12 Capsure Z 4533) and 37 ventricular (24 Capsure SP 4023 and 13 Capsure Z 4033) implanted in 47 consecutive patients (30 men and 17 women, with an age of 72 +/- 9.4 years). The pacemaker model was Medtronic Elite 7077-7086 (DDD-DDDR) in 25 patients and Medtronic Legend 8419-8424 (VVIR-AAIR) in 22 patients. The endocavitary signal (all patients had spontaneous rhythm) was telemetrically obtained by a Medtronic 9790 device and acquired on a personal computer at implantation and 7, 30, and 180 days thereafter. The signal was studied both in the time domain and in the frequency domain by spectral analysis. The following parameters were calculated: amplitude (A): peak-to-peak value of the complex; slew rate (SR) peak negative first derivative; F0: frequency at which the power spectrum reaches its maximum value; and bandwidth (Bw): expressed as the distance between the -3 dB points and statistically analyzed by a two-way analysis of variance with factors "time" (four measurements) and "electrode" (Capsure SP and Z) and repeated measurements on the former. Ventricular sensing: no time or electrode effect (P > 0.1 in all comparisons) was found for F0, Bw, or SR, while a time effect (P < 0.04) not dependent on the type of electrode was found for the amplitude of the signal. In particular, a significant increase was found between the measurement at 6 months and that at implantation (P < 0.004). Atrial sensing: A, F0, and bandwidth were not affected by time or electrode (P > 0.09), while SR behaved differently over time (P < 0.05) in the two electrodes (the Capsure Z showed an increase at sixth month [P < 0.04] compared to implantation). In conclusion, the Medtronic Capsure SP and Z electrodes proved to be valid and substantially equivalent as far as concerns the measurement of the intracardiac potential despite the difference between their surface areas. Further studies should be devised to assess whether transitory decreases of atrial Bw in the first month of follow-up observed in a few patients for both electrodes could be responsible for clinical episodes of sensing deficit. PMID- 10461293 TI - Head-up tilt test in patients with high pretest likelihood of neurally mediated syncope: an approximation to the "real sensitivity" of this testing. AB - This study was designed to examine the "true sensitivity" of a specific head-up tilt (HUT) testing protocol using clinical findings. The HUT protocol used 45 minutes at 60 degrees for the baseline portion and intermittent boluses of 2, 4, and 6 micrograms of isoproterenol in the second phase. Eighty-eight patients (40 men and 48 women; mean age of 33.8 +/- 16 years) with recurrent syncope and high pretest likelihood of neurally mediated syncope were included. The following were considerated as high pretest likelihood criteria: (1) at least two syncopal episodes; (2) no structural heart disease and normal baseline ECG; (3) age < 65 years; (4) a typical history of neurally mediated syncope, triggering factors plus premonitory signs; and (5) short duration of symptoms and fast recovery without neurological sequelae. Fifty-four patients (61%) had a positive tilt test (34/88 baseline [39%] and 20/50 with isoproterenol [40%]). The shorter time interval between the last syncopal episode and baseline HUT test was the only predictor for a positive response (P < 0.003). Conversely, this time interval was not predictor of positive responses during isoproterenol-tilt testing. IN CONCLUSION: (1) we claim a "sensitivity" for this combined protocol of 61%; and (2) our results indicate that patients with syncope of unknown origin must be tilted nearest as possible to the last syncope to increase the positive responses of HUT test. PMID- 10461294 TI - Can early timed internal atrial defibrillation shocks reduce the atrial defibrillation threshold? AB - The defibrillation threshold is markedly reduced very early following the initiation of ventricular fibrillation. The purpose of this study was to determine if the same finding holds true for atrial defibrillation. Sustained, reproducible AF was induced with programmed atrial pacing using acetyl-beta methylcholine chloride (40-640 microL/min) in six adult sheep (heart weight 245 300 g). Seven timing intervals (125 ms, 200 ms, 1 s, 3 s, 10 s, 30 s, and 5 min after AF induction) and two lead configurations: (1) RA as cathode and CS as anode; and (2) RA as cathode and RV apex as anode were tested. Single capacitor biphasic waveforms (3/1 ms) were delivered and atrial defibrillation thresholds (ADFTs) were determined in random order. No significant differences in leading edge voltage and total energy were detected for the RA-CS configuration for the seven timing intervals. For the RA-RV configuration, a significant difference was detected comparing the voltage for 125 ms to the 5-minute timing interval. For all times except 125 ms, the RA-RV threshold was significantly higher than the RA CS level. In contrast to ventricular defibrillation, the ADFT does not change significantly within the first 5 minutes after the initiation of AF for the RA-CS configuration. However, if the shock is given very early (125 ms after AF induction) with the RA-RV configuration, the ADFT is lowered almost to the RA-CS level. PMID- 10461295 TI - Defibrillation thresholds are increased by right-sided implantation of totally transvenous implantable cardioverter defibrillators. AB - Whether an ICD is placed via a left- or right-sided approach depends on venous access, the presence of a preexisting pacemaker, and other factors. Since the DFT is affected by lead position, which in turn is determined in part by the side of access, right-sided venous access could adversely affect DFTs. Furthermore, right sided active can placement directs electric current toward the right hemithorax, which could further increase DFTs. This study sought to determine whether DFTs were increased by right-sided vascular access, and whether active can technology was beneficial or detrimental with right-sided ICD placement. Stepdown to failure DFTs were found in 290 patients receiving transvenous systems at the time of initial ICD implantation. Of these, 271 (93%) received left-sided systems and 19 (7%) received right-sided systems. The mean DFT in systems placed via left-sided vascular access was 11.3 +/- 5.3 J versus 17.0 +/- 4.9 J for right-sided implantation (P < 0.0001); right-sided DFTs were elevated for both active can and cold can systems. Right-sided active can devices had a lower DFT than right-sided cold can systems (15 +/- 4.1 J vs 19 +/- 4.8 J, P = 0.05). The right-sided implantation of implantable defibrillators results in significantly higher DFTs than the left-sided approach. This may be due to the less favorable distribution of the defibrillating field relative to the myocardium with the devices on the right. When right-sided implantation is clinically mandated, active can devices result in lower thresholds and should be used. PMID- 10461296 TI - Permanent pacing in patients with univentricular heart. AB - Patients with one of several varieties of malformation designated as "univentricular hearts" may be especially challenging when permanent pacing is required. Our objective was to review our experience in this subset of patients that had undergone permanent pacing and thus determine the optimal approach. A retrospective chart review was done of 32 patients with some variety of "univentricular" malformation who had required permanent pacing at our institution. Although technically challenging, permanent pacing in this group of patients can be successful through several approaches. The various approaches, as well as consideration of the differences that exist between patients undergoing septation and those undergoing a Fontan procedure are discussed. Although long term permanent pacing is possible in this group of patients, before pacing begins, a thorough understanding of the anatomy and prior surgical procedures is necessary. Use of a combined atrial transvenous and ventricular epicardial pacing system may work well for some patients. With the development of newer and more reliable coronary sinus leads, dual chamber transvenous pacing with ventricular stimulation via the coronary sinus could become the approach of choice in some patients with "univentricular hearts." PMID- 10461297 TI - Single center experience with femoral extraction of permanent endocardial pacing leads. AB - Between March 1995 and June 1997, 128 leads were extracted from the hearts of 28 women and 50 men, 69 +/- 15 years of age (mean +/- SD, range 22-92 years). The indications for the procedure were: Accufix leads in 18 patients (14%), dysfunction or incompatibility with ICD in 16 (12%), endocarditis on the lead in 41 (32%), pulse generator pocket infection in 28 (22%), and pulse generator and/or lead erosion in 25 patients (19%). The extraction was performed with a snare (lasso), via a femoral vein as a first approach in 116 leads, and as an alternate approach, after extraction from the original site of implantation had failed, in 12 leads. The leads had been implanted for 62 +/- 48 months (range 1 205 months). A Cook sheath was used in 7, and a femoral approach traction in 20 instances. Of the 128 leads, 122 (95%) were completely extracted, and 2 (2%) were partially extracted (the distal electrode remaining attached to the myocardium), and 4 (3%) could not be removed. Four complications occurred: 2 tears of the tricuspid valve without clinical consequences, one separation of the lead's distal electrode which migrated into the hypogastric vein, and one hemorrhage at the femoral puncture site. There was no death or serious complication caused by lead extraction in this series. PMID- 10461298 TI - Nonfluoroscopic localization of an amagnetic stimulation catheter by multichannel magnetocardiography. AB - This study was performed to: (1) evaluate the accuracy of noninvasive magnetocardiographic (MCG) localization of an amagnetic stimulation catheter; (2) validate the feasibility of this multipurpose catheter; and (3) study the characteristics of cardiac evoked fields. A stimulation catheter specially designed to produce no magnetic disturbances was inserted into the heart of five patients after routine electrophysiological studies. The catheter position was documented on biplane cine x-ray images. MCG signals were then recorded in a magnetically shielded room during cardiac pacing. Noninvasive localization of the catheter's tip and stimulated depolarization was computed from measured MCG data using a moving equivalent current-dipole source in patient-specific boundary element torso models. In all five patients, the MCG localizations were anatomically in good agreement with the catheter positions defined from the x-ray images. The mean distance between the position of the tip of the catheter defined from x-ray fluoroscopy and the MCG localization was 11 +/- 4 mm. The mean three dimensional difference between the MCG localization at the peak stimulus and the MCG localization, during the ventricular evoked response about 3 ms later, was 4 +/- 1 mm calculated from signal-averaged data. The 95% confidence interval of beat-to-beat localization of the tip of the stimulation catheter from ten consecutive beats in the patients was 4 +/- 2 mm. The propagation velocity of the equivalent current dipole between 5 and 10 ms after the peak stimulus was 0.9 +/- 0.2 m/s. The results show that the use of the amagnetic catheter is technically feasible and reliable in clinical studies. The accurate three-dimensional localization of this multipurpose catheter by multichannel MCG suggests that the method could be developed toward a useful clinical tool during electrophysiological studies. PMID- 10461299 TI - Electrophysiological and clinical comparison of two temporary pacing leads following cardiac surgery. AB - Temporary pacing leads are invaluable in diagnosis and treatment of arrhythmia after cardiac surgery. The ideal lead possesses reliable pacing and sensing capabilities throughout the postoperative period. Ease of handling and a low complication rate are essential. This study compares a new lead (Medtronic model #6492) to a temporary pacemaker lead established in clinical use (Medtronic model #6500). One lead of each type was placed in the right atrium and ventricle in 33 patients undergoing elective coronary revascularization. Pacing function was measured on the first and fourth postoperative days. Lead complications were documented. On day 1, there was no statistical difference between lead types in terms of pacing (voltage threshold, current threshold), impedance, or sensing (P wave amplitude, R wave amplitude). On day 4, both leads showed an increase in pacing threshold and a decrease in sensing ability. The only statistical difference between lead types was in atrial sensing on day 4, as measured by the P wave amplitudes (1.95 +/- 0.18 V for model 6492 vs 1.40 +/- 0.14 V for model 6500, P < 0.05). Two leads of each model failed to pace in the ventricular position and one lead in each model in the atrial position. There were no complications attributable to either pacemaker lead. We found that both lead types were reliable and functioned well. The Medtronic Model #6492 lead demonstrated better long-term sensing, although in this study the difference was not clinically significant. PMID- 10461300 TI - Five-year follow-up of a bipolar steroid-eluting ventricular pacing lead. AB - Steroid-eluting pacing leads are known to attenuate the threshold peaking early after implantation. Long-term performance, however, is not yet settled. The lead design tested in this prospective study combines a 5.8-mm2 tip of microporous platinum-iridium with elution of 1.0 mg of dexamethasone sodium phosphate and tines for passive fixation (model 5024, Medtronic Inc.). In 50 patients (mean age 69 +/- 10 years), the electrode was implanted in the right ventricular apex. Follow-up was performed on days 0, 2, 5, 10, 28, 90, 180 and every 6 months thereafter for 5-years postimplant. At each visit, pacing thresholds were determined as pulse duration (ms) at 1.0 V and as the minimum charge (microC) delivered for capture. Lead impedance (omega) was telemetered at 2.5 V-0.50 ms, and sensing thresholds (mV) were measured in triplicate using the automatic sensing threshold algorithm of the pacemaker implanted (model 294-03, Intermedics Inc.). On the day of implantation, mean values were 0.10 +/- 0.03 ms, 0.12 +/- 0.03 microC, 758 +/- 131 omega, and 13.1 +/- 1.8 mV, respectively. Beyond 1-year postimplant, pacing thresholds did not vary significantly. Sensing thresholds and lead impedance values were stable during long-term follow-up. Five years after implantation, mean values were 0.23 +/- 0.11 ms, 0.24 +/- 0.07 microC, 670 +/- 139 omega, and 11.6 +/- 3.1 mV for pulse width and charge threshold, lead impedance, and sensing threshold, respectively, and all leads captured at 1.0 V with the longest pulse duration available (1.50 ms). It is concluded that the bipolar steroid-eluting tined ventricular lead showed stable stimulation thresholds, lead impedance values, and sensing thresholds for 5 years after implantation. PMID- 10461301 TI - The effect of biphasic defibrillation on the immediate pacing threshold of a dedicated bipolar, steroid-eluting lead. AB - It is apparent that pacing threshold increases following an ICD shock, although the degree of change observed is dependent on the method used to assess pacing and the lead design used. We previously demonstrated a rise in postshock pacing threshold using a lead with integrated bipolar pacing in which the distal shocking coil also serves as the pacing anode. In this study, we sought to investigate whether the postshock pacing threshold increased significantly in an endocardial, steroid-eluting lead with dedicated bipolar pacing electrodes. Twenty patients (16 men, 4 women; median age 73, ejection fraction [EF] 0.17 0.58) were studied during pectoral ICD implantation (Medtronic active can model 7221Cx or 7223Cx with model 6932-65 lead). The diastolic pulse width pacing threshold at 1 or 2 V was determined. Pacing rate was set > or = 100/min at twice diastolic threshold output to assess pacing immediately following the first DFT test shock. For subsequent shocks, the output was adjusted to establish postshock thresholds as 1, 2, 3, or 4 times the diastolic threshold. The postshock threshold was defined as the output yielding 100% capture > or = 2.5 seconds following a shock. In 8 of 20 patients (ratio 0.40 +/- 0.11), a rise in the post shock threshold was shown by failure of consistent capture when pacing at 2 times diastolic threshold > or = 2.5 seconds after a DFT test shock. Two of these patients failed at 3 times threshold, but none failed at 4 x threshold. Five of 12 patients with successful capture of 2 times threshold failed to capture at threshold. The postshock threshold increased by a mean factor of 2.83 +/- 0.83 in the group of patients with a threshold rise. Following ICD shock in an active can, steroid-eluting lead system with dedicated bipolar pacing, the post-shock threshold increases significantly. Our studies suggest a need for postshock pacing to be set at least 4 x threshold regardless of the lead design. PMID- 10461302 TI - A comparison of ventricular function during high right ventricular septal and apical pacing after his-bundle ablation for refractory atrial fibrillation. AB - This study compares LV performance during high right ventricular septal (RVS) and apical (RVA) pacing in patients with LV dysfunction who underwent His-bundle ablation for chronic AF. We inserted a passive fixation pacing electrode into the RVA and an active fixation electrode in the RVS. A dual chamber, rate responsive pulse generator stimulated the RVA through the ventricular port and the RVS via the atrial port. Patients were randomized to initial RVA (VVIR) or RVS (AAIR) pacing for 2 months. The pacing site was reversed during the next 2 months. At the 2 and 4 month follow-up visit, each patient underwent a transthoracic echocardiographical study and a rest/exercise first pass radionuclide ventriculogram. We studied nine men and three women (mean age of 68 +/- 7 years) with congestive heart failure functional Class (NYHA Classification): I (3 patients), II (7 patients), and III (2 patients). The QRS duration was shorter during RVS stimulation (158 +/- 10 vs 170 +/- 11 ms, P < 0.001). Chronic capture threshold and lead impedance did not significantly differ. LV fractional shortening improved during RVS pacing (0.31 +/- 0.05 vs 0.26 +/- 0.07, P < 0.01). RVS activation increased the resting first pass LV ejection fraction (0.51 +/- 0.14 vs 0.43 +/- 0.10, P < 0.01). No significant difference was observed during RVS and RVA pacing in the exercise time (5.6 +/- 3.2 vs 5.4 +/- 3.1, P = 0.6) or the exercise first pass LV ejection fraction (0.58 +/- 0.15 vs 0.55 +/- 0.16, P = 0.2). The relative changes in QRS duration and LV ejection fraction at both pacing sites showed a significant correlation (P < 0.01). We conclude that RVS pacing produces shorter QRS duration and better chronic LV function than RVA pacing in patients with mild to moderate LV dysfunction and chronic AF after His bundle ablation. PMID- 10461303 TI - Dynamic beat-to-beat QT-RR relationship during physiotherapy effort in elderly patients without primary heart disease. AB - The ECGs from 18 patients hospitalized in a rehabilitation setting, following surgery for hip fracture, were examined to characterize the dynamic behavior of uncorrected QT interval in relation to changing RR interval during physiotherapy effort. ECG waveforms were analyzed to extract beat-to-beat QT and RR intervals using a computerized ECG Analyzer (CEA-1100). The method of defining the QT and RR intervals is based on performing multiple cross-correlations that enable rejection of artifacts from the analysis. The relationship between the RR and QT intervals was found using the following general formula QTi = cRRi-1b. Linear regression was performed on the logarithms of QT and RR measurements obtained to estimate the constant (a = log c) and the slope (b) values, reflecting the dynamic change of QT during physiotherapy effort. Having these two values, the dynamic QT extrapolated to a heart period of 1 second (QTcd) was calculated. The results were compared to the conventional corrected static QT according to the Bazzet formula (QTcs). The mean values of constants (a = log c) and slopes (b) over all patients were found to be 1.61 +/- 0.23 and 0.33 +/- 0.08, respectively, giving a QT (ms) heart-period (ms) dynamic relation of QTi = 41 x RR(i-1)0.33. The correlation between the dynamic QT and the static QT intervals was not significant. The mean values of the QTcd and QTcs intervals were significantly different (392 +/- 25 ms vs 434 +/- 28 ms; P < 0.0001). This dynamic measurement method of QT intervals may provide additional information on normal and abnormal cardiac repolarization in health and disease, helping in the diagnosis of cardiac disorders and arrhythmia risk. PMID- 10461304 TI - An unusual ECG from a patient with an atriobiventricular pacemaker. PMID- 10461305 TI - Pacing and ICD therapy in Germany (West and East) before and after reunification in 1989. AB - In 1989 the two German countries, FRG and GDR, were reunited after 50 years of political separation. During this time, these countries underwent independent, and in parts quite different, developments. While the reunification has had less effect on the overall situation in the Western part of the country, the Eastern part has experienced considerable changes, including the health service. In the field of pacing and ICD therapy, this study finds that 8 years after the reunification, the German federal states of both parts of the country have converged in the field of pacing and ICD therapy. PMID- 10461306 TI - Heartbeat international: a program designed to use pacemakers as peacemakers. PMID- 10461307 TI - Asymptomatic form of Brugada syndrome. AB - We describe a patient with the asymptomatic form of Brugada syndrome. His electrographical, electropharmacological, and electrophysiological characteristics were similar to those reported in patients with the symptomatic form of Brugada syndrome. We believe that he has the same arrhythmogenic substrate as that of patients with Brugada syndrome. The fact that he had no episode of spontaneous ventricular fibrillation might be explained by his absence of the triggering factors. PMID- 10461308 TI - Brugada-like electrocardiographic pattern in a patient with a mediastinal tumor. AB - We report on a patient with a mediastinal tumor and electrocardiographic findings similar to those described in the Brugada syndrome. This peculiar ECG pattern disappeared after tumor removal, thus suggesting it was probably caused by compression of the right ventricular outflow tract by the mass. PMID- 10461309 TI - Ibutilide for termination of atrial fibrillation in the Wolff-Parkinson-White syndrome. AB - Ibutilide promptly restored sinus rhythm on two occasions in an elderly patient with AF and rapid ventricular response associated with the WPW syndrome. As a selective Class III antiarrhythmic agent that prolongs cardiac refractoriness, ibutilide offers an alternative effective therapy for rapid termination of AF in WPW. PMID- 10461310 TI - Cough-induced heart block. PMID- 10461311 TI - Peripheral pulmonary migration of a retained pacemaker lead. PMID- 10461312 TI - Trends toward subcutaneous prepectoral implantable cardioverter defibrillator (ICD) implantation. PMID- 10461313 TI - Association of sunlight exposure and photoprotection measures with clinical outcome in systemic lupus erythematosus. AB - This study was designed to explore the relationship of sunlight exposure and ultraviolet (UV) light protection measures with clinical outcome in systemic lupus erythematosus (SLE). A structured questionnaire was administered to sixty Puerto Rican SLE patients, to assess their attitudes and behavior regarding sunlight exposure and photoprotection measures. Medical records were reviewed to evaluate the clinical outcome measures that included: clinical manifestations, number of SLE-related hospitalizations, number of exacerbations and pharmacologic treatment. Almost all (98.3%) patients were well acquainted of sunlight effects on disease activity. Two thirds were exposed to direct sunlight for an average of less than one hour per day and 33.3% for one hour or more. Thirty patients (50%) reported use of sunscreen, with sun protective factor of 15 or greater, when exposed to sunlight. Less than 40% of patients regularly wore hat or long-sleeves clothes to protect from sunlight. Although there were some clinical differences between the groups with different sunlight exposure times, none reached statistical significance. Also, no significant differences were found between the groups in regards to sunlight protective clothes. However, patients that regularly used sunscreen had significantly lower renal involvement (13.3 vs 43.3%), thrombocytopenia (13.3 vs 40%), hospitalizations (26.7 vs. 76.7%), and requirement of cyclophosphamide treatment (6.7 vs. 30%) than patients that did not used it (P < 0.05). We conclude that use of sunscreen photoprotection was associated with a better clinical outcome in our SLE patients. These findings further support the importance and benefits of photoprotective measures in patients with SLE. PMID- 10461314 TI - Malignant melanoma in Puerto Rico: an update. AB - A study about the incidence of malignant melanoma in Puerto Rico during the calendar year 1996 was carried out compiling the information submitted by pathological reports to the Puerto Rico Cancer Registry and identifying similar reports at other pathology laboratories. A total of 107 new cases were documented in 1996, resulting in an incidence of 3.03 (+/- 17.9) per 100,000 persons for that particular year. The age of the patients fluctuated between 19 and 88 years with a mean of 61.3 years, most of them being between 50 and 80 years. More than half of the cases (54.2%) were located in the head and neck, and the extremities. Thickness measurement of the neoplasm was reported in only 42% of the cases. In 60% of those cases in which the tumor thickness was reported, it was found to be less than 1.49 mm. Previous data in Puerto Rico from the years 1981 to 1986 had shown a tendency to an increase in the incidence of melanoma per 100,000 inhabitants rising 1.20 to 1.52 respectively. The present study reflects a continuation of the same pattern, with a two-fold increase in the last 10 years (from 1.52 to 3.03). PMID- 10461315 TI - Diagnosis of malaria by polymerase chain reaction. AB - Malaria is no longer endemic in Puerto Rico, however, imported cases of the disease are occasionally reported to the Health Department of the Island. This is a report of a 45-year-old female patient who traveled to Kenya and Niger and was admitted to a San Juan area hospital with an 8 day history of daily chills and fever, myalgia, nausea and vomiting. Upon admission, peripheral blood displayed multiple intra-erythrocytic ring-shape trophozoites, highly suggestive of Plasmodium falciparum. The polymerase chain reaction was used as a complementary method for the detection of malaria parasites and confirmation of post-treatment parasite clearance. This report presents an imported case of malaria in Puerto Rico and showed the use of a molecular technique to diagnose Plasmodium. PMID- 10461316 TI - A tail of protein folding. AB - This review describes the use of a simple genetic system that has provided important insight into the process of folding and, of its flipside, that of protein aggregation. These studies make use of the tail protein of the bacterial virus P22 which infects Salmonella typhimurium. This folding system serves as a model for a number protein structural elements and may also provide important insights into disease-related protein folding defects at a time when an increasing number of diseases are being shown to be due to protein folding alterations. PMID- 10461317 TI - An epidemiological review of tuberculosis in the Puerto Rican population. AB - Past and recent developments related with tuberculosis (TB) epidemiology in the island were reviewed. Characteristics of the disease such as surveillance, incidence, mortality and the relation of TB with HIV infection were included. TB remains an important public health problem in Puerto Rico and in the Americas, particularly, in HIV-infected patients. From 1981 through 1998, a total of 1,000 AIDS/TB cases has been reported in the island accounting for 4.4% of the total of AIDS cases. 82% of the AIDS/TB cases were males. The incidence was highest among injecting drug users (IDUs) infected with HIV. The overall incidence of TB in recent years seems to reach a plateau. By the contrary, AIDS/TB cases have been increasing. However, problems of under-reporting must be taken into consideration when analyzing morbidity trends. Adequate and innovative solutions to maintain TB control in the community must be promptly implemented. PMID- 10461318 TI - The history of cardiovascular surgery in Puerto Rico. PMID- 10461319 TI - Fine-needle aspiration diagnosis of hydatid cyst. AB - The diagnosis of hydatid disease outside endemic areas is usually not suspected. Hydatid cysts in imaging studies can be confused with hepatic tumors, abscesses, cystadenomas, liver cysts or other lesions. Serology is the usual confirmatory test, but cytologic diagnosis has been described. Aspiration of the cysts has not been employed as a routine diagnostic method for fear of spillage and anaphylactic reactions. We report a case of unsuspected hepatic echinococcosis that was confirmed by fine-needle aspiration of the lesion and cytologic confirmation without complications. PMID- 10461320 TI - [Bioethical considerations in the treatment of terminal patients]. PMID- 10461321 TI - [Clinical practice guidelines on the Internet]. PMID- 10461322 TI - Prenatal cellular transplantation. AB - Based on the ontogeny of fetal immunologic development, the strategy of fetal cellular transplantation may prove to be the most physiological way to achieve replacement of abnormal hemoglobin, immune cells, or enzyme defects diagnosed early prenatally. Similarly, if cellular transplantation can induce tolerance, postnatal solid organ transplantation may be performed without the need for chronic immunosuppression or with a reduced risk of graft-versus-host disease. This review presents experimental data from the late 1940s until the present for both small and large animal models; it also describes the limited clinical experience with prenatal cellular transplantation. PMID- 10461323 TI - Molecular approaches to understanding organogenesis. AB - The elucidation of the molecular mechanisms of mammalian organogenesis is the foundation on which we can build an improved understanding of organ pathology and pathophysiology. This paper uses the lung and the pancreas as paradigms to demonstrate how advances in basic molecular developmental biology research has translated into new appreciation of, and even novel potential treatment strategies for, congenital anomalies and mature diseases of these organs. PMID- 10461324 TI - The current status of tissue engineering as potential therapy. AB - End-stage organ disease and tissue loss continue to be major medical problems. Although transplantation has become an established and successful method of therapy, the severe scarcity of donor organs, especially in the pediatric population, has become a major limitation and has stimulated investigation into selective cell transplantation. The authors have been investigating the fabrication of functional living tissue, or tissue engineering, using cells seeded on highly porous synthetic biodegradable polymer scaffolds as a novel approach toward the development of biological substitutes that may replace lost tissue function. Over the past decade, we have applied the principles of tissue engineering in the fabrication of a wide variety of tissues, including both structural and visceral organs. This article reviews the progress that has been achieved and the current status of tissue engineering as potential therapy for end-stage organ disease and tissue loss. PMID- 10461325 TI - Fetal wound repair: where do we go from here? AB - In contrast to adult wound healing, early-gestation fetal skin wound healing occurs rapidly, in a regenerative fashion, and without scar formation. The accelerated rate of healing, relative lack of an acute inflammatory response, and an absence of neovascularization distinguishes fetal from adult wound healing. However, this remarkable ability of the fetus to heal without scarring still remains poorly understood. The uncertainties include the role of cytokines, extracellular matrix components, homeobox genes, and certain cell types in the scarless wound repair process. Nevertheless, some strides have been made within the last two decades. This report, discusses the current knowledge of the mechanisms and characteristics of scarless fetal wound healing. Furthermore, to shy away from being just another all inclusive review, the authors point out deficiencies in the knowledge base on this important topic. Last, the future direction of research is discussed that may elucidate the mechanisms regulating the scarless repair phenomena. PMID- 10461326 TI - The metabolic needs of critically ill children and neonates. AB - The pediatric metabolic response to injury and operation is proportional to the degree of stress and causes an increase in the turnover of proteins, fats, and carbohydrates. Thereby, substrates are made readily available for the immune response and wound healing. Because this process requires energy, the resting energy expenditure of ill patients increases. Whole-body protein degradation rates are elevated out of proportion to synthetic rates, and negative protein balance also ensues. Neonates and children are particularly susceptible to the loss of lean body mass and its attendant increased morbidity and mortality caused by an intrinsic lack of endogenous stores and greater baseline requirements. An appropriately designed mixed fuel system of nutritional support replete in protein does not quell this metabolic response but can result in anabolism and continued growth in ill children. In addition, the use of adequate analgesia and anesthesia is a readily available and proven means of reducing the magnitude of the catabolism associated with operation and injury. Finally, as hormonal- and cytokine-mediated metabolic alterations are better understood, therapeutic interventions may become available to directly modulate the metabolic response to illness, thus potentially further improving clinical outcome in pediatric surgical patients. PMID- 10461327 TI - New approaches to understanding the etiology and treatment of total parenteral nutrition-associated cholestasis. AB - Total parenteral nutrition-associated cholestasis (TPN-AC) may be a fatal disease. The only known effective treatment is to discontinue TPN and institute full enteral feedings. However, this is not possible for many patients with severe gastrointestinal failure. Current research supports two theories regarding the etiology of TPN-AC. One proposes that the enteral fast disrupts the enterohepatic circulation. Cholestasis, in this hypothesis, results from a combination of altered gut hormone production and endotoxins produced by bacterial translocation. The second theory implicates the direct toxicity of TPN solution. Amino acid solutions and plant sterols in intralipid have generated much interest. Ursodeoxycholic acid and S-adenosyl-L-methionine are promising treatments for TPN-AC. They have been proven to be effective in animals and adult liver diseases. Cholecystokinin also has been investigated as a possible prophylactic agent. However, results from these experiments do not conclusively show a beneficial effect. PMID- 10461328 TI - Nitric oxide and intestinal barrier failure. AB - The systemic inflammatory response syndrome (SIRS) is a leading cause of morbidity and mortality in adults and children. Various proinflammatory mediators have been implicated in the pathogenesis of SIRS; however, their mechanisms of action are poorly defined. Recent evidence suggests that nitric oxide (NO) plays a regulatory role in gut barrier function. Sustained upregulation of NO production in the intestine can lead to intestinal epithelial injury through the formation of peroxynitrite. Peroxynitrite can nitrate mitochondrial proteins and inhibit cellular respiration. The resultant changes in mitochondrial function lead to activation of the caspase cascade, subsequent DNA fragmentation, and enterocyte apoptosis. Enterocyte apoptosis results in a transient "bare area" in the intestinal epithelium where bacteria can attach and then penetrate the lamina propria. Bacteria that successfully escape the immune system may in turn incite a systemic inflammatory response. PMID- 10461329 TI - Respiratory failure: current status of experimental therapies. AB - A number of advances in the treatment of infants and children with respiratory failure have been investigated in the laboratory with translation to clinical practice. Investigators have recognized that application of high ventilating pressures and failure to apply adequate levels of positive end-expiratory pressure (PEEP) can inflict injury to the already failing lung. Other interventions such as prone positioning and application of new ventilating strategies such as proportional assist ventilation (PAV), inverse ratio ventilation (IRV), high frequency ventilation, liquid ventilation, and intratracheal pulmonary ventilation (ITPV), continue to be developed and explored. Administration of inhaled nitric oxide (iNO) may improve pulmonary physiology and gas exchange in patients with respiratory insufficiency. Finally, the technique of extracorporeal life support (ECLS) is being simplified and refined. This report summarizes the status of these advances and describes the basic science and clinical research that brought them to clinical application. PMID- 10461330 TI - Prenatal ultrasonographic diagnosis of nuchal cord(s): disregard, inform, monitor or intervene? PMID- 10461331 TI - Ultrasound dating of pregnancy--still controversial? PMID- 10461332 TI - Fetal outcome in pregnancies defined as post-term according to the last menstrual period estimate, but not according to the ultrasound estimate. AB - OBJECTIVE: To study the risk of adverse fetal outcome in fetuses that were post term according to the last menstrual period estimate but not according to the ultrasound estimate. DESIGN: A total of 11,510 women with singleton pregnancies, reliable last menstrual period and delivery after 37 weeks were divided into four groups: women who delivered at term, i.e. within 259-295 days according to both the ultrasound and the last menstrual period estimate; women who delivered post term according to the last menstrual period estimate but not according to the ultrasound estimate; women who delivered post-term according to the ultrasound estimate but not according to the last menstrual period estimate; and women who delivered post-term according to both the ultrasound and the last menstrual period estimates. Stepwise logistic regression was used to test whether the risk of Apgar score of < 7 after 5 min and transfer to the neonatal intensive care unit increased in any of the post-term groups. RESULTS: There was no significant difference in mortality between the term group and the three study groups. There was no significant increase in the risk for Apgar score of < 7 after 5 min or transfer to the neonatal intensive care unit for pregnancies that were defined as post-term according to the last menstrual period estimate but not according to the ultrasound estimate. There was, however, an increased risk for Apgar score of < 7 after 5 min in the group that was post-term according to the ultrasound estimate but not according to the last menstrual period estimate. There was also an increased risk for transfer to the neonatal intensive care unit in the group that was post-term according to both estimates. CONCLUSION: The effect of ultrasound in changing the estimated day of delivery to a later date leading to pregnancies becoming post-term according to the last menstrual period estimate but not according to the ultrasound estimate does not have any adverse consequences for the fetal outcome. However, there seems to be an increased risk for adverse consequences for pregnancies that are post-term according to the ultrasound estimate. PMID- 10461333 TI - Fetal outcome when the ultrasound estimate of the day of delivery is more than 14 days later than the last menstrual period estimate. AB - OBJECTIVE: To evaluate the effect on fetal outcome of changing the estimated day of delivery as calculated according to ultrasound measurements more than 14 days later than the day estimated according to the last menstrual period. DESIGN: A non-selected population comprising 15,241 women was evaluated. A study group (the day of delivery based on the ultrasound estimate being changed to more than 14 days later than the estimate based on the last menstrual period) and a control group (the two estimates being within 7 days of each other) were compared regarding various parameters concerning fetal outcome. RESULTS: Changing the estimated day of delivery, based on the ultrasound evaluation, to a date 14 days later than the day of delivery as estimated according to the last menstrual period did not influence the risk of abortion, perinatal death or transferral to the neonatal intensive care unit. There was a difference of 3 days in the accuracy of the prediction of day of delivery between the two groups. There was a greater number of infants with a birth weight below 2500 g in the study group, but no difference was found between the groups in the number of infants with a birth weight < 2 SD from the mean according to the ultrasound estimate. CONCLUSION: There was no indication of any adverse consequence of the routine scan and change of estimated day of delivery among 15,000 pregnancies in a non selected population. PMID- 10461335 TI - Tetralogy of Fallot in the fetus: findings at targeted sonography. AB - OBJECTIVES: To evaluate the findings of tetralogy of Fallot in various fetal sonographic views. METHODS: We reviewed the fetal sonograms and medical records of 20 fetuses with prenatal diagnosis of tetralogy of Fallot. We analyzed the indications for targeted sonography, the abnormalities seen in various sonographic views, the postnatal echocardiographic and angiographic findings and autopsy findings. RESULTS: The most common indication for targeted sonography was an abnormal (n = 12) or inadequate (n = 3) finding on sonographic screening in which the abnormality was most frequently found on the three-vessel view (n = 9). The key pathological features of tetralogy of Fallot were uniformly demonstrated in the ventricular outflow tract, three-vessel and short-axis views. The ductus arteriosus was small in 70% of cases and not identifiable in the remaining fetuses. In three of six fetuses with no identifiable ductus, the ductus was shown to be absent at autopsy. The direction of ductal flow was variable. CONCLUSION: The key features of tetralogy of Fallot were always demonstrable in the ventricular outflow tract, three-vessel and short-axis views. The most common reason for referral was the abnormal three-vessel view. PMID- 10461334 TI - Evaluation of ultrasound-estimated date of delivery in 17,450 spontaneous singleton births: do we need to modify Naegele's rule? AB - OBJECTIVE: To compare the size of error in the predicted date of delivery by biparietal diameter (BPD) and last menstrual period (LMP) in different clinical models. MATERIALS AND METHODS: Predicted dates of delivery were calculated in 14,805 spontaneous deliveries with a reliable date of LMP using two assumptions: average length of pregnancy of 280 and of 282 days. Errors in these calculated dates were tested when used alone or combined, i.e. the LMP-predicted date of delivery was used as first choice unless the discrepancy between gestational age calculated by BPD and LMP exceeded 7, 10 or 14 days. RESULTS: The average discrepancy (error) between predicted date of delivery from BPD and LMP and date of spontaneous delivery was 7.96 and 8.63 days, respectively (p < 0.0001). Adding 282 instead of 280 days to the first day of the LMP reduced the error of the LMP method from 8.63 to 8.41 days, reduced the percentage of classified post-term deliveries from 7.9 to 5.2% and increased the preterm births from 3.96 to 4.48%. No models of combined use of LMP and BPD were superior to the use of BPD alone. CONCLUSION: If both BPD and LMP are available, BPD should be used to predict term. If only LMP is available, term should be calculated as the first date of the LMP plus 282. PMID- 10461336 TI - Comparison of the non-stress test with the evaluation of centralization of blood flow for the prediction of neonatal compromise. AB - OBJECTIVE: To determine whether the middle cerebral to umbilical artery systolic/diastolic velocity waveform ratio (MC/UA) was a more sensitive indicator of fetal compromise than the non-stress test (NST). STUDY DESIGN: An outcome study of high-risk patients undergoing NST testing and MC/UA studies within 10 days of delivery. Patients were divided into four groups based on their test results, and neonatal outcome parameters were compared. RESULTS: There were significant differences between all four test result groups with respect to length of stay in the neonatal intensive care unit (NICU). Patients in whom both the NST and MC/UA ratio were normal had significantly lower utilization of Cesarean section for delivery, admission and length of stay in the NICU and occurrence of significant neonatal complications. Logistic regression analysis indicated that the combination of the NST and MC/UA ratio was an excellent predictor of perinatal outcome. CONCLUSIONS: The MC/UA ratio improves the sensitivity for the prediction of poor perinatal outcome when it is combined with the NST. PMID- 10461337 TI - Prenatal assessment of Wharton's jelly in umbilical cords with single artery. AB - OBJECTIVE: To investigate whether the amount of Wharton's jelly in non-malformed fetuses with a single umbilical artery is different from that of fetuses with a normal umbilical cord. METHODS: We evaluated patients with singleton pregnancies, non-malformed fetuses and single umbilical artery undergoing sonographic evaluation at a gestational age ranging from 19 to 41 weeks' gestation. The cross sectional areas of the umbilical cord and of the umbilical vessels were measured. The amount of Wharton's jelly was calculated by subtracting from the total cross sectional area of the umbilical cord the areas of the artery and of the vein. The umbilical cord cross-sectional area, the umbilical artery and vein areas as well as the amount of Wharton's jelly were plotted on previously published nomograms. RESULTS: Twenty-two patients met the inclusion criteria. The umbilical cord cross sectional area was within the normal range in 20 (90.1%) cases. The umbilical artery and vein areas were above 2 standard deviations from the mean in 20 cases and in 11 cases (50%), respectively. The amount of Wharton's jelly was below 2 standard deviations from the mean in all cases. An abnormal insertion of the umbilical cord (marginal, velamentous) was present in five cases (22.7%). CONCLUSIONS: A reduction of Wharton's jelly is frequently present in cases of single umbilical artery. The increased perinatal morbidity and mortality observed in cases of single umbilical artery, even in the absence of congenital or chromosomal abnormalities, could be in part the consequence of a reduced amount of Wharton's jelly. PMID- 10461338 TI - The possible impact of a myomatous uterus on the accuracy of fetal biometric measurements. AB - OBJECTIVE: The propagation speed of ultrasound beams changes in different media. The aim of the study was to assess the possible measurement error of fetal biometry performed in cases of a myomatous uterus. DESIGN: Three iron phantoms of 30 mm, 50 mm and 70 mm were measured by ultrasound. Each phantom was measured by three different transducers of 3.5 MHz, 6.5 MHz and 7.5 MHz. Measurements were performed in a water bath and the intraobserver variability was assessed. Thereafter phantoms were measured with one of their edges covered by a specimen of a myomatous uterus while their other edge was covered with water. RESULTS: Artifactual lengthening of the phantoms' size was observed in measurements performed with the uterine myoma. The measurement errors were inversely related to the length of the phantoms and became statistically significant in the phantoms of 30 mm (13.15 +/- 3.16% without the myoma vs. 17.38 +/- 4.39% with the myoma; p < 0.0001). Measurement errors were independent of the transducer type (p = 0.001, p = 0.01 and p = 0.014 for the 3.5-MHz, 6.5-MHz and 7.5-MHz transducers, respectively). CONCLUSIONS: Ultrasound measurement errors are expected while examining small fetal organs which are partially overshadowed by a myoma. PMID- 10461339 TI - The effect of the menstrual cycle on human cerebral blood flow: studies using Doppler ultrasound. AB - OBJECTIVE: Previous studies have demonstrated hemodynamic changes at different phases in the menstrual cycle, but the cerebral circulation has not been investigated. Our aim was to study carotid and cerebral blood flow during the menstrual cycle using Doppler ultrasound. Two different techniques of Doppler waveform analysis were used: standard Doppler indices and Laplace transform analysis (LTA), which may provide additional hemodynamic information. DESIGN: This was a prospective study of healthy volunteers who were providing pre conception data for a subsequent longitudinal study set in the Department of Obstetrics and Gynaecology, Nottingham University Hospital. Nineteen women were studied in the mid-follicular and mid-luteal phases of 27 ovulatory menstrual cycles. Doppler recordings were obtained from the internal and external carotid and middle cerebral arteries. The standard Doppler indices (systolic/diastolic ratio, pulsatility index and resistance index) and LTA parameters were calculated. RESULTS: The standard Doppler indices were all significantly higher in the luteal compared to the follicular phase in the right middle cerebral artery (p < 0.05). However, no changes were seen in the standard indices in the carotid arteries or in any of the LTA parameters in any artery. Using the LTA, vessel wall stiffness was greater and absolute velocity of flow lower in the middle cerebral compared to the carotid arteries. CONCLUSIONS: Increased ventilation and a subsequent lowering of alveolar CO2 pressure secondary to a raised progesterone level in the mid-luteal phase could account for the observed changes within the middle cerebral artery. Under the conditions of this study the LTA appears less sensitive at detecting alterations in downstream resistance compared to the standard Doppler indices. PMID- 10461340 TI - The value of sonohysterography combined with cytological analysis of the fluid retrieved from the endometrial cavity in predicting histological diagnosis. AB - OBJECTIVE: The purpose of the study was to assess the efficacy of sonohysterography combined with cytological analysis of the fluid retrieved from the endometrial cavity in predicting histological diagnosis. STUDY DESIGN: A prospective study was conducted comparing sonohysterography combined with endometrial washings for cytology with histological evaluation after surgical procedures. Of 152 patients referred for sonohysterography, 87 were premenopausal and 65 were postmenopausal. Some of the injected fluid was aspirated for cytological analysis. Sixty-one patients (40%) underwent surgical hysteroscopy and eight (5%) had dilatation and curettage as a result of the sonohysterographic findings. Histological diagnoses were compared with the sonohysterographic and cytological findings. RESULTS: In 99 (65%) patients, sonohysterography demonstrated endometrial polypoid lesions. Only 54 endometrial cavitary lesions were confirmed pathologically. Epithelial cells with atypia were more often found in patients without (five of 53) than in those with (two of 99) an endometrial polyp (p < 0.05). Only one out of nine cases of histological diagnosis of hyperplasia was predicted cytologically. CONCLUSIONS: The addition of cytological analysis of the fluid retrieved from the endometrial cavity during sonohysterography did not contribute to the prediction of benign histological diagnosis of endometrial hyperplasia. PMID- 10461341 TI - Perinatal management of fetal hemolytic disease due to Rh incompatibility combined with fetal alloimmune thrombocytopenia due to HPA-5b incompatibility. AB - We report out experience in the perinatal management of a complex case of fetal hemolytic disease primarily due to Rhesus incompatibility combined with fetal alloimmune thrombocytopenia. The lowest fetal hemoglobin and platelet levels were 2.6 g/dl and 13,000/microliter, respectively. Intrauterine treatment consisted of six transfusions of packed red cells into the umbilical vein and one transfusion of platelets. The neonate required four transfusions of packed red cells to correct her hyporegenerative erythropoiesis. Postnatal management also included one platelet transfusion, intravenous immunoglobulins and erythropoietin. Although some degree of fetal thrombocytopenia may invariably be found in fetal red cell incompatibility, other rare causes need to be excluded. PMID- 10461342 TI - Pathophysiology and treatment of fetal anemia due to placental chorioangioma. AB - Placental chorioangiomas occur in 1% of pregnancies. Large chorioangiomas may cause serious complications such as fetal anemia, hydrops and fetal death. In this case report, a pregnancy complicated by a large placental chorioangioma is described. Severe fetal anemia without the occurrence of hydrops fetalis was suspected using ultrasound and Doppler examinations. Successful intrauterine blood transfusion was performed, with an unusually large amount of blood needed to obtain an adequate rise in fetal hematocrit. Two weeks later, at 32 weeks, the infant was born in good condition. In pregnancies with large chorioangiomas, we advise regular ultrasound and Doppler examinations, with the aim of detecting fetal anemia before hydrops develops. When anemia is suspected, fetal blood sampling is indicated and intrauterine transfusion therapy may be beneficial to preserve fetal health until maturity is reached. PMID- 10461343 TI - Prenatal diagnosis of dacryocystocele: a possible marker for syndromes. AB - We present our experience of the sonographic prenatal diagnosis of dacryocystocele and review the literature. This lesion can be distinguished from a facial hemangioma, dermoid or anterior encephalocele by the ultrasound findings and Doppler flow studies. These conditions are separate entities and their associated diagnosis and prognosis are very different in each case. As dacryocystocele may be part of numerous syndromes, its prenatal visualization raises the rare possibility of associated anomalies. PMID- 10461344 TI - Prenatal sonographic diagnosis of dacryocystocele. PMID- 10461345 TI - Picture of the month. Evolution of umbilical cord entanglement in monoamniotic twins. PMID- 10461346 TI - Ultrasound-induced morphological changes in the murine small intestine. PMID- 10461347 TI - Gastric neuroendocrine cells and secretory products. AB - The ECL cell is the most common cell type in the oxyntic mucosa of the stomach. It is producing a number of peptides and amines where histamine and chromogranin A seems to be the most important and abundant products. Recent data indicate a direct correlation between ECL-cell mass and circulating chromogranin A levels. Chromogranin A and its splice products might serve as growth promoting agents in ECL-cell hyperplasia or gastric carcinoids. PMID- 10461348 TI - Ontogeny of ECL cells in the rat. AB - ECL cells produce histamine and chromogranin A, and are restricted to the oxyntic mucosa of the stomach. ECL cell ontogeny has been studied in some detail in the rat. Using histidine decarboxylase immunostaining, the first ECL cells can be demonstrated at embryonic day 17. Immunoreactive histamine and chromogranin A appear one day later. At embryonic day 20, the vesicular monoamine transporter type 2 is also present in the ECL cells. Neonatally the ECL cell proliferation is slow; however, one to three weeks postnatally there is a rapid growth of ECL cells to populate the basal half of the glands. Gastrin is known to be an important stimulator of ECL cell activity and growth in the adult rat. As revealed in recent mouse gene knock out models gastrin does not seem to play a role in the early ECL cell differentiation and development. PMID- 10461349 TI - Physiology of the ECL cells. AB - The enterochromaffin-like (ECL) cells of the oxyntic mucosa (fundus) of the stomach produce, store and secrete histamine, chromogranin A-derived peptides such as pancreastatin, and an unanticipated but as yet unidentified peptide hormone. The cells are stimulated by gastrin and pituitary adenylate cyclase activating peptide and suppressed by somatostatin and galanin. Choline esters and histamine seem to be without effect on ECL cell secretion. The existence of a gastrin-ECL cell axis not only explains how gastrin stimulates acid secretion but also may help to explore the functional significance of the ECL cells with respect to the nature and bioactivity of its peptide hormone. From the results of studies of gastrectomized/fundectomized and gastrin-treated rats, it has been speculated that the anticipated ECL-cell peptide hormone acts on bone metabolism. PMID- 10461350 TI - The fate of released histamine: reception, response and termination. AB - Histamine released from ECL cells elicits responses from a variety of cellular targets in the vicinity. Three sets of receptors are involved (H1, H2 and H3). Receptor occupation is promptly transduced into cellular responses. The responses, in turn, are terminated by diverse mechanisms: enzymatic inactivation, cellular uptake and desensitization at the receptor level. Under specific pathological conditions, histamine effects could be exaggerated by the presence of derivatives that may be of marginal relevance under physiological conditions. PMID- 10461351 TI - Physiological significance of ECL-cell histamine. AB - In the oxyntic mucosa of the mammalian stomach, histamine is stored in ECL cells and in mucosal mast cells. In the rat, at least 80 percent of oxyntic mucosal histamine resides in the ECL cells. Histamine is a key factor in the regulation of gastric acid secretion. Following depletion of ECL-cell histamine by treatment with alpha-fluoromethylhistidine (alpha-FMH), basal acid secretion was reduced, and gastrin-stimulated acid secretion was abolished. Vagally-induced acid secretion (by insulin injection or pylorus ligation) was unaffected by alpha-FMH treatment but inhibited by an H2 antagonist. These results suggest that gastrin stimulates acid secretion via release of ECL-cell histamine, whereas vagally induced acid secretion--although histamine-dependent--does not rely on ECL-cell histamine. Gastrin is known to have a trophic effect on the oxyntic mucosa. By combining long-term hypergastrinemia with continuous infusion of alpha-FMH, we were able to show that gastrin-evoked trophic effects in the stomach do not depend on ECL-cell histamine. PMID- 10461352 TI - Control of histidine decarboxylase gene expression in enterochromaffin-like cells. PMID- 10461353 TI - Histamine metabolism of gastric carcinoids in Mastomys natalensis. AB - Pharmacological inhibition of gastric acid secretion and subsequent hypergastrinemia in Mastomys natalensis is an experimental model well suited for the study of gastric carcinoid formation. The genetic susceptibility of Mastomys to develop such tumors is a feature reminiscent of the situation in patients with the MEN-1 Zollinger Ellison syndrome, in whom tumor-induced hypergastrinemia, promotes the development of gastric carcinoids. Chronic hypergastrinemia, induced by the irreversible H2-receptor antagonist loxtidine will cause carcinoid formation in Mastomys already after four to six months. As in humans, gastric carcinoids in Mastomys are mainly composed of enterochromaffinlike (ECL) cells and have low malignant potential. Administration of exogenous gastrin to normal young animals increases the expression of histidine decarboxylase (HDC) mRNA in the oxyntic mucosa within 30 minutes. Endogenous hypergastrinemia, induced by short-time loxtidine treatment (three to 29 days) enhances the expression of HDC mRNA, histamine contents and ECL cell numbers in the oxyntic mucosa. Long-term loxtidine treatment (seven to 21 months) results in sustained hypergastrinemia and tumor formation. Tumor-bearing animals exhibited an increase in HDC mRNA and histamine content in the oxyntic mucosa as well as increased urinary excretion of the main histamine metabolite, tele-methylimidazole acetic acid (MeImAA). Subsequent to cessation of loxtidine treatment for two weeks, all parameters of histamine metabolism were normalized in tumor-bearing animals. These results indicate that gastric carcinoids developing during hypergastrinemia are well differentiated neoplasms whose histamine synthesis and metabolism is regulated by plasma gastrin. PMID- 10461354 TI - ECL cell morphology. AB - Using immunohistochemistry at the conventional light, confocal and electron microscopic levels, we have demonstrated that rat stomach ECL cells store histamine and pancreastatin in granules and secretory vesicles, while histidine decarboxylase occurs in the cytosol. Furthermore the ECL cells display immunoreactivity for vesicular monoamine transporter type 2 (VMAT-2), synaptophysin, synaptotagmin III, vesicle-associated membrane protein-2, cysteine string protein, synaptosomal-associated protein of 25 kDa, syntaxin and Munc-18. Using electron microscopy in combination with stereological methods, we have evidence to suggest the existence of both an exocytotic and a crinophagic pathway in the ECL cells. The process of exocytosis in the ECL cells seems to involve a class of proteins that promote or participate in the fusion between the granule/vesicle membrane and the plasma membrane. The granules take up histamine by VMAT-2 from the cytosol during transport from the Golgi zone to the more peripheral parts of the cells. As a result, they turn into secretory vesicles. As a consequence of stimulation (e.g., by gastrin), the secretory vesicles fuse with the cell membrane to release their contents by exocytosis. The crinophagic pathway was studied in hypergastrinemic rats. In the ECL cells of such animals, the secretory vesicles were found to fuse not only with the cell membrane but also with each other to form vacuoles. Subsequent lysosomal degradation of the vacuoles and their contents resulted in the development of lipofuscin bodies. PMID- 10461355 TI - Properties of isolated gastric enterochromaffin-like cells. AB - The gastric enterochromaffin-like cell (ECL) has been studied in gastric fundic glands by confocal microscopy and as a purified cell preparation by video imaging of calcium signaling and measurements of histamine release. Regulation of gastric acid secretion is largely due to alterations of histamine activation of the H2 receptor on the parietal cell and can be divided into central neural regulation, with direct actions of neuronally released mediators and into peripheral regulation by substances released from other endocrine cells. Gastric neuronal stimulation of acid secretion by alteration of ECL cell function is probably mediated by pituitary adenylate cyclase activating peptide (PACAP) receptors on the ECL cell, which activate calcium signaling and histamine release. Peripheral stimulation of acid secretion via the ECL cell is largely mediated by gastrin stimulation of calcium signaling and histamine release. Gastric neuronal inhibition of ECL cell function is probably mediated by galanin inhibition of calcium signaling, and histamine release and peripheral inhibition of ECL cell function is mainly due to somatostatin release from D cells. PMID- 10461356 TI - The role of endogenous gastrin in the development of enterochromaffin-like cell carcinoid tumors in Mastomys natalensis: a study with the specific gastrin receptor antagonist AG-041R. AB - We examined the effects of a newly synthesized gastrin receptor antagonist, AG 041R, on the growth of enterochromaffin-like (ECL) carcinoid tumors in Mastomys natalensis both in vitro and in vivo. AG-041R was as potent as the well known gastrin antagonist L365,260 in inhibiting not only the gastrin-induced release of histamine from but also histidine decarboxylase (HDC) gene expression in the ECL carcinoid tumor cells. AG-041R also inhibited gastrin-induced DNA synthesis and c fos gene expression in the tumor cells. Furthermore, AG-041R significantly inhibited the growth of the transplanted Mastomys ECL carcinoid tumors in vivo. From these data, it is concluded that endogenous gastrin is involved in the growth of ECL carcinoid tumors in Mastomys natalensis. Moreover, AG-041R is shown to have a potential as an anti-neoplastic agent for ECL carcinoid tumor of the stomach. PMID- 10461357 TI - Recognition and the histopathologic classification of ECL cell proliferations. PMID- 10461358 TI - Pathogenesis of ECL cell tumors in humans. AB - In ECL cell tumors developed in the setting of hypergastrinemic conditions (ECL cell carcinoids type 1 and 2), hypergastrinemia is the dominant agent acting as a promoter in all steps (hyperplasia-dysplasia-neoplasia) of the tumorigenic sequence. In contrast, it apparently lacks transforming properties as shown by the absence of ECL cell carcinoids in patients exposed to hypergastrinemia alone, i.e., those with sporadic Zollinger-Ellison syndrome. The potential transforming factors include: the allelic loss of the MEN-1 suppressor gene in the genetically predisposed MEN-1 patients, an alteration that may induce ECL cell tumors even in the absence of hypergastrinemia; the still unknown factor(s) associated with atrophic corporal gastritis; agents whose role in the induction of human ECL cell tumors is still unclarified, such as basic Fibroblast Growth Factor, human Chorionic Gonadotropin-alpha and Transforming Growth Factor-alpha; and agents having a favoring role on the ECL exposure to mitogens such as BCL-2. No information is currently available on the pathogenesis of gastrin-independent, sporadic ECL cell carcinoids (type 3) or of gastric neuroendocrine carcinomas. PMID- 10461359 TI - Natural history, clinicopathologic classification and prognosis of gastric ECL cell tumors. AB - A series of 50 gastric endocrine tumors classified according to Rindi et al. [1] comprised 12 small cell neuroendocrine carcinomas (NEC) and 38 ECL cell carcinoids, of which 22 associated with type A chronic atrophic gastritis (A CAG), eight with hypertrophic gastropathy due to combined Multiple Endocrine Neoplasia and Zollinger/Ellison syndrome (MEN/ZES), and eight sporadic. Variables found to predict tumor malignancy were: size > 2 cm, > 2 mitoses and > 130 Ki67 positive cells/10 high power fields (HPF), grade 2 or 3 histology, angioinvasion, p53 protein nuclear accumulation, and the presence of a single tumor. None of these factors increased significantly the predicting ability of tumor classification itself, although grade 2 + 3 shows 100 percent negative predictive value and Ki67 and angioinvasion 100 percent positive predictive value. When the mostly non-malignant A-CAG and MEN-ZES tumors were analysed against the mostly malignant sporadic and NEC tumors, a positive predictive value of 90 percent and a negative predictive value of 93 percent was obtained. Investigation of a larger tumor series is under way with the aim to develop an optimal model for prognostic evaluation of gastric endocrine tumors. PMID- 10461360 TI - Hypergastrinemia and enterochromaffin-like cell hyperplasia. AB - The enterochromaffin-like (ECL) cells, the most frequent endocrine cells of the oxyntic mucosa of the stomach, are under the trophic stimulus of gastrin. These cells undergo a hyperplastic increase in variety of hypergastrinemic diseases. The most widely accepted nomenclature for the description of hyperplastic proliferation has been retrospectively arranged in a sequence presumed to reflect a temporal evolution of the proliferative process. A comparative, prospective study aimed to verify, in human hypergastrinemic diseases such as atrophic body gastritis (ABG), Zollinger-Ellison syndrome (ZES) and antral gastrin cell hyperfunction (AGCH), the effect of exposure of ECL cells to different pattern of gastrin hypersecretion, is lacking. To this purpose, we studied a series of consecutive patients with ABG, ZES and AGCH at the time of first diagnosis. MATERIAL AND METHODS: The patients included in this study (124 ABG, 18 ZES and 10 AGCH) were selected on the basis of two previously performed screening studies aimed to diagnose these diseases. All patients at the time of diagnosis underwent gastroscopy, with multiple biopsies of the gastric body mucosa for the evaluation of qualitative pattern of ECL cells hyperplasia, and basal fasting gastrin determination. A sample of hypergastrinemic patients from each group was further investigated by meal-stimulation of gastrin secretion and quantitative morphometry for CgA positive gastric body endocrine cells. RESULTS: AGCH patients showed only the normal or simple hyperplasia pattern. In the ZES group, simple and linear grades accounted for 38.4 percent and 46.1 percent, respectively. MEN I patients showed only these two patterns. The majority of ABG patients showed the presence of micronodular pattern (59.7 percent). A correlation analysis between fasting gastrin levels and grade of hyperplasia (r = 0.5580, p < 0.0001), indicates that the greater the gastrin levels, the higher is the degree of severity of ECL hyperplasia pattern. In conclusion, our data support the role of gastrin as the selective contributor to the progression of ECL cell hyperplasia in humans. PMID- 10461361 TI - Clinical aspects of ECL-cell abnormalities. AB - ECL cell hyperplasia results from hypergastrinemia, and in man this occurs due to achlorhydria in atrophic gastritis (pernicious anemia [PA]) and gastrinoma (Zollinger-Ellison syndrome [ZES]). Progression to neoplasia, i.e., ECL cell carcinoids (usually small, multicentric and non-functional), occurs in some five to 10 percent of patients with PA where they remain gastrin-dependent and reversible by normalization of serum gastrin by antrectomy. Even if untreated, the carcinoids are almost invariably benign and do not cause death. In ZES, ECL cell hyperplasia is progressive due to hypergastrinemia. However, carcinoids develop only in the MEN-I subtype but pose no additional threat of malignancy. A conservative approach is recommended for small multicentric carcinoids, and the tumors do not need removal. By contrast, single, large, non-gastrin-dependent carcinoids represent a different biological and clinical problem and are frequently malignant. PMID- 10461362 TI - Diagnosis and treatment of ECL cell tumors. AB - The diagnosis of ECL-omas is easy to perform. In patients with Zollinger-Ellison syndrome (ZES), ECL-omas are almost always observed in the setting of multiple endocrine neoplasia type I. In patients without ZES, the first step is to discard non-gastrin-related sporadic ECL-omas whose prognosis is poor. By contrast, prognosis of ECL-omas in patients with ZES or chronic atrophic gastritis is good. Metastases are rare, and tumor-related deaths are exceptional. In both conditions, ECL-omas measuring less than 1 cm should be treated by endoscopic polypectomy and survey. Treatment modalities (surgery, endoscopic polypectomy) for larger tumors are still discussed. The impact of endoscopic ultrasonography on the therapeutic decision has not yet been evaluated. Considering the good prognosis of these tumors, aggressive surgery could be limited to selected patients. Multicentric studies should be undertaken to determine the best treatment modalities. PMID- 10461363 TI - Relationship of ECL cells and gastric neoplasia. AB - The enterochromaffin-like (ECL) cell in the oxyntic mucosa has a key role in the regulation of gastric secretion since it synthesizes and releases the histamine regulating the acid secretion from the parietal cell. Gastrin is the main regulator of the ECL cell function and growth. Long-term hypergastrinemia induces ECL cell hyperplasia, and if continued, neoplasia. ECL cell carcinoids occur in man after long-term hypergastrinemia in conditions like pernicious anemia and gastrinoma. There is also accumulating evidence that a proportion of gastric carcinomas of the diffuse type is derived from the ECL cell. Furthermore, the ECL cell may, by producing substances with angiogenic effects (histamine and basic fibroblast growth factor), be particularly prone to develop malignant tumors. Although the general opinion is that gastrin itself has a direct effect on the oxyntic mucosal stem cell, it cannot be excluded that the general trophic effect of gastrin on the oxyntic mucosa is mediated by histamine or other substances from the ECL cell, and that the ECL cell, therefore, could play a role also in the tumorigenesis/carcinogenesis of gastric carcinomas of intestinal type. PMID- 10461364 TI - Small synthetic ligands of the cholecystokinin-B/gastrin receptor can mimic the function of endogenous peptide hormones. AB - The gastric cholecystokinin-B/gastrin receptor (CCK-BR) is a key regulator of enterochromaffin-like cell function and proliferation. Over the last decade, a number of small non-peptide CCK-BR "antagonists" have been discovered. Here, we demonstrate that some of these non-peptide ligands in fact possess significant ability to activate the human CCK-BR, and are, therefore, more properly categorized as partial agonists. When tested in COS-7 cells transiently expressing the recombinant human CCK-BR, saturating concentrations of the small "peptoid" ligands PD 135,158 and PD 136,450 stimulated inositol phosphate formation to 23 and 43 percent, respectively, of the maximum response induced by a considerably larger endogenous peptide agonist, cholecystokinin octapeptide. In contrast, the benzodiazepine-derived CCK-BR ligand, YM022, acted as a "true" high affinity antagonist of cholecystokinin-induced inositol phosphate formation (pA2 = 9.69). Consistent with recent findings in animal experiments, our data reveal that small synthetic ligands have the potential to function as either CCK-BR agonists or antagonists. These dual properties of synthetic molecules must be considered when evaluating candidate drugs for human disease. PMID- 10461365 TI - Abnormal gastric morphology and function in CCK-B/gastrin receptor-deficient mice. AB - Mice lacking the cholecystokinin (CCK)-B/gastrin receptor have been generated by targeted gene disruption. The roles of this receptor in controlling gastric acid secretion and gastric mucosal growth have been assessed. The analysis of homozygous mutant mice vs. wild type included measurement of basal gastric pH, plasma gastrin concentrations as well as quantification of gastric mucosal cell types by immunohistochemistry. Mutant mice exhibited a marked increase in basal gastric pH (from 3.2 to 5.2) and about a 10-fold elevation in circulating carboxyamidated gastrin compared with wild-type controls. Histologic analysis revealed a decrease in both parietal and enterochromaffin-like (ECL) cells, thus explaining the reduction in acid output. Consistent with the elevation in circulating gastrin, antral gastrin cells were increased in number while somatostatin cells were decreased. These data support the importance of the CCK B/gastrin receptor in maintaining the normal cellular composition and function of the gastric mucosa. PMID- 10461366 TI - Biological processing of fossil fuels. Resume of the Bioconversion Session of ICCS'97 PMID- 10461367 TI - Biotechnology and microbiology of coal degradation AB - For several years it has been known that fungi and bacteria can attack and even liquefy low rank coals. This review covers the progress in coal biotechnology and microbiology, mainly during the last decade, from describing the first effects to elucidating the mechanisms used by the microorganisms. More than one mechanism is responsible for microbial coal degradation/liquefaction: oxidative enzymes (peroxidases, laccases), hydrolytic enzymes (esterases), alkaline metabolites and natural chelators. Due to the heterogeneous structure of coal, which is described in one section, and for economic reasons the review focuses on the enzymatic depolymerization of brown coal. Approaches which seem not so promising are discussed (anaerobic, reductive pathways, chemical pretreatment). Finally the possible applications and products in this field are summarized, as lignite with a worldwide production of about 940 million tons a year will continue to play an important economic role in the future. PMID- 10461368 TI - Fungal biosolubilization of rhenish brown coal monitored by Curie-point pyrolysis/gas chromatography/mass spectrometry using tetraethylammonium hydroxide AB - Residues and coal fractions that remained after the biosolubilization of Rhenish brown coal by strains of Lentinula edodes and Trametes versicolor have been studied by Curie-point pyrolysis/gas chromatography/mass spectrometry using tetraethylammonium hydroxide (NEt4OH) at 610 degrees C. To differentiate methyl derivatives of esters and ethers from free or bound hydroxyl and carboxyl groups NEt4OH was used in the thermochemolysis experiments instead the commonly used tetramethylammonium hydroxide. A comparison of humic acid fractions before and after fungal attack shows considerable alteration of the soluble macromolecules of coal. Depending on the coal fraction studied and the fungi used, the assortment of fatty acid esters released during the pyrolysis varies significantly. Furthermore, dicarbonic acid ethyl diesters as well as ethyl derivatives of aromatic ethers and acids yield information about humic acid structure and the biosolubilization of brown coal. Variations in the mixture produced are possibly caused by differences in the pattern of extracellular enzymes secreted that attack the macromolecular structural elements of brown coal. Therefore pyrolysis of native and microbiologically altered geomacromolecules using NEt4OH allows one to differentiate between free hydroxyl groups as well as substances that are attached to humic substances via ester or ether bridges, and their methylated counterparts. PMID- 10461369 TI - Processes of liquefaction/solubilization of Spanish coals by microorganisms AB - Several fundamental aspects of microbial coal liquefaction/solubilization were studied. The liquefied/solubilized products from coal by microorganisms were analysed. The liquid products analysed by IR titration and UV/visible spectrometry showed some alterations with regard to the original coal. Humic acids extracted from the liquefied lignite showed a reduction in the average molecular weight and a increase in the condensation index, probably due to depolymerization caused by microorganisms. The mechanisms implicated in coal biosolubilization by two fungal strains, M2 (Trichoderma sp.) and M4 (Penicillium sp.) were also studied. Extracellular peroxidase, esterase and phenoloxidase enzymes appear to be involved in coal solubilization. PMID- 10461370 TI - Mechanisms of coal solubilization by the deuteromycetes Trichoderma atroviride and Fusarium oxysporum AB - Three different mechanisms can be envisaged that are used by fungi to solubilize coal: the production of alkaline substances, the extrusion of chelators and, of special interest in the scope of biotechnology, the action of enzymes. Whether these mechanisms are operating separately or in various combinations has not yet been finally assessed. The two deuteromycetes Fusarium oxysporum and Trichoderma atroviride solubilize coal by synergistic effects of various differnt mechanisms depending on the cell metabolism. F. oxysporum seems to solubilize coal by increasing the pH of the mycelial surroundings and by the action of chelators induced during growth in glutamate-containing media (without involvement of enzymes). T. atroviride, on the other hand, appears to use, in addition to an alkaline pH and a high chelator activity, at least two classes of enzyme activity to attack coal: hydrolytic activity for coal solubilization and ligninolytic activity for degradation of humic acids. PMID- 10461371 TI - Evidence for and expression of a laccase gene in three basidiomycetes degrading humic acids. AB - The majority of lignin-degrading basidiomycetes are able to depolymerize humic acids. In this presentation the relationship and possible similarities between enzymes involved in lignin degradation and humic acid depolymerization were examined on the genetic level. We have cloned fragments of the gene encoding the extracellular ligninolytic enzyme laccase from Clitocybula dusenii, Nematoloma frowardii and a fungal strain designated i63-2, and compared the three sequences with those of several other published laccase genes. The sequenced fragments displayed a high homology both on the DNA (97%-77%) and amino acid (100%-85%) level. Furthermore, the expression of this gene in the above-mentioned fungi was demonstrated by a nested polymerase chain reaction with cDNA as template. PMID- 10461372 TI - Transformation of macromolecules from a brown coal by lignin peroxidase AB - Indirect evidence has suggested that lignin peroxidase (LiP) of the white-rot fungus Phanerochaete chrysosporium catalyses oxidative decolourisation and depolymerisation of macromolecules from brown coal in vivo. In this study we show that LiP catalyses these transformations in vitro. Unmethylated (USC45 coal) and methylated (MWSC6 coal) fractions of solubilised macromolecules (M(r) > 30,000) from a brown coal were treated with a semi-purified preparation of LiP isozymes from P. chrysosporium. Both coal fractions were decolourised, losing between 26% and 39% of their absorbance at both 280 nm and 400 nm, in reactions that had an absolute requirement for H2O2 and veratryl alcohol. Neither coal fraction was transformed when the enzyme was heat-inactivated or in the presence of the LiP inhibitor metavanadate. Gel-permeation chromatography showed that MWSC6 coal but not USC45 was depolymerised and yielded low-molecular-mass (M(r) < 30,000) fragments. Nine monomeric products were identified by GC-MS. PMID- 10461373 TI - Degradation of lignite (low-rank coal) by ligninolytic basidiomycetes and their manganese peroxidase system AB - Ligninolytic basidiomycetes (wood and leaf-litter-decaying fungi) have the ability to degrade low-rank coal (lignite). Extracellular manganese peroxidase is the crucial enzyme in the depolymerization process of both coal-derived humic substances and native coal. The depolymerization of coal by Mn peroxidase is catalysed via chelated Mn(III) acting as a diffusible mediator with a high redox potential and can be enhanced in the presence of additional mediating agents (e.g. glutathione). The depolymerization process results in the formation of a complex mixture of lower-molecular-mass fulvic-acid-like compounds. Experiments using a synthetic 14C-labeled humic acid demonstrated that the Mn peroxidase catalyzed depolymerization of humic substances was accompanied by a substantial release of carbon dioxide (17%-50% of the initially added radio-activity was released as 14CO2). Mn peroxidase was found to be a highly stable enzyme that remained active for several weeks under reaction conditions in a liquid reaction mixture and even persisted in sterile and native soil from an opencast mining area for some days. PMID- 10461375 TI - Biosynthesis of polyhydroxyalkanoates from low-rank coal liquefaction products by Pseudomonas oleovorans and Rhodococcus ruber AB - A screening identified several bacteria that were able to use chemically heterogeneous low-rank coal liquefaction products as complex carbon sources for growth. Pseudomonas oleovorans and Rhodococcus ruber accumulated polyhydroxyalkanoic acids (PHA) amounting to 2%-8% of the cell dry weight when the cells were cultivated on these liquefaction products in the absence of any other carbon source. R. ruber accumulated, in addition to PHA, small amounts of triacylglycerols. The accumulated PHA consisted of 3-hydroxyhexanoate, 3 hydroxydecanoate, and 3-hydroxydodecanoate (P. oleovorans) or 3-hydroxybutyric acid and 3-hydroxyvaleric acid (R. ruber). Low-rank coal liquefaction products obtained from Trichoderma atroviride were better substrates for P. oleovorans than chemically produced fulvic acids. PMID- 10461374 TI - Degradation of alicyclic molecules by Rhodococcus ruber CD4. AB - The present work describes investigations on the bacterial degradation of the alicyclic molecule cyclododecane. It represents a structure where the initial degradative steps have to be similar to a "subterminal" attack as there is no "terminal" part of the molecule. We were able to show that the gram-positive bacterium Rhodococcus ruber CD4 DSM 44394 oxidizes cyclododecane to the corresponding alcohol and ketone, the latter being subject to ring fission by a Baeyer-Villiger oxygenase. This key enzyme is an NADPH- and O2-dependent flavoprotein with a substrate specificity for bigger rings. The further metabolism of the resulting lactone gives rise to an omega-hydroxyalkanoic acid that is susceptible to common beta-oxidation. Due to its alicyclic character and its ring size, cyclododecane is comparable to aliphatic bridge components that are an important element in the coal texture. They contribute to the three dimensional coal structure and thus could serve as a valuable target for the oxidative abilities of R. ruber CD4 to reduce the molecular mass of coal. PMID- 10461376 TI - Developments in destructive and non-destructive pathways for selective desulfurizations in oil-biorefining processes AB - Biocatalytic desulfurization is still not a commercial technology, but conceptual engineering and sensitivity analyses have shown that the approach is very promising. The purpose of this paper is to investigate further some aspects of the biodesulphurization pathways, discussing the non-destructive pathway with the well-known Rhodococcus rhodochrous IGTS8. Findings revealed byproducts, such as 2'-hydroxybiphenyl (HBP), sulfite and sulfate, obtained by the desulfurization of dibenzothiophene (DBT), to exert an inhibiting effect. The results suggest that IGTS8 may follow two different metabolic pathways in stationary-growth-phase cells or under growing conditions. The first pathway is characterized by oxidative steps, which convert DBT to DBT sulfoxide and to DBT sulfone. The sulfone is transformed to 2-(2'-hydroxyphenyl)benzene sulfinate and then to HBP and sulfite by a sulfinic acid hydrolase. In the second pathway the sulfone is further oxidized to 2-(2'-hydroxyphenyl)benzene sulfonate and then to HBP and sulfate by a sulfonic acid hydrolase. Experiments using benzene sulfonic acid suggest that the sulfonic acid hydrolase is an induced enzyme. PMID- 10461377 TI - Bioremoval of organic and inorganic sulphur from coal samples AB - The microbial ecology of different Spanish coal samples has been studied. Several bacteria have been isolated from enrichment cultures and characterised and their biodesulphurization abilities evaluated. Using morphological and physiological properties, different isolates have been related to species of the Xanthomonas, Pseudomonas, Chryseomonas and Moraxella genera. Some of the isolates, B(30)15 and T(30)10, gave important levels of organic desulphurization, close to 70%. Other isolates, B(30)7 and B(30)8, were able to remove inorganic sulphur with high efficiencies, over 67%. One of the isolates, B(30)10, metabolically related to Xanthomonas maltophila, was able to remove both organic and inorganic sulphur at neutral pH, with efficiencies of 69% and 68% respectively. The results obtained underline the potential use of some of these strains for industrial coal desulphurization processes. PMID- 10461378 TI - Manipulation of the DNA coding for the desulphurizing activity in a new isolate of Arthrobacter sp. AB - A new bacterial strain able to cleave C-S bonds from organosulphur heterocyclic compounds through the 4-S pathway and tentatively classified as Arthrobacter sp. was recently isolated. In the present short article we describe the cloning and the characterization of the DNA encoding the enzymes responsible for desulphurization in this microorganism, referred to as Arthrobacter sp. DS7. The desulphurization operon was found to be located in a large plasmid that also bears the genes conferring cadmium and arsenic resistance. By shortening this plasmid, a new cloning vector was prepared and used to obtain a recombinant derivative strain that desulphurizes dibenzothiophene despite of the presence of inorganic sulphur in the growth medium. PMID- 10461379 TI - Bioassays for risk assessment of coal conversion products. AB - Traditional as well as biotechnological processing of coal leads to complex mixtures of products. Besides chemical and physical characterization, which provides the information for product application, there is a need for bioassays to monitor properties that are probably toxic, mutagenic or cancerogenic. Investigations carried out focused on the selection, adaptation and validation of bioassays for the sensitive estimation of toxic effects. Organisms like bacteria, Daphnia magna and Scenedesmus subspicatus, representing different complexities in the biosphere, were selected as test systems for ecotoxicological and mutagenicity studies. The results obtained indicate that bioassays are, in principle, suitable tools for characterization and evaluation of coal-derived substances and bioconversion products. Using coal products, coal-relevant model compounds and bioconversion products, data for risk assessment are presented. PMID- 10461380 TI - A comparative genomics approach to the evolution of eukaryotes and their mitochondria. AB - The Organelle Genome Megasequencing Program (OGMP) investigates mitochondrial genome diversity and evolution by systematically determining the complete mitochondrial DNA (mtDNA) sequences of a phylogenetically broad selection of protists. The mtDNAs of lower fungi and choanoflagellates are being analyzed by the Fungal Mitochondrial Genome Project (FMGP), a sister project to the OGMP. Some of the most interesting protists include the jakobid flagellates Reclinomonas americana, Malawimonas jakobiformis, and Jakoba libera, which share ultrastructural similarities with amitochondriate retortamonads, and harbor mitochondrial genes not seen before in mtDNAs of other organisms. In R. americana and J. libera, gene clusters are found that resemble, to an unprecedented degree, the contiguous ribosomal protein operons str, S10, spc, and alpha of eubacteria. In addition, their mtDNAs code for an RNase P RNA that displays all the elements of a bacterial minimum consensus structure. This structure has been instrumental in detecting the rnpB gene in additional protists. Gene repertoire and gene order comparisons as well as multiple-gene phylogenies support the view of a single endosymbiotic origin of mitochondria, whose closest extant relatives are Rickettsia-type alpha-Proteobacteria. PMID- 10461381 TI - Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. AB - Small subunit rRNA sequence data were generated for 27 strains of cyanobacteria and incorporated into a phylogenetic analysis of 1,377 aligned sequence positions from a diverse sampling of 53 cyanobacteria and 10 photosynthetic plastids. Tree inference was carried out using a maximum likelihood method with correction for site-to-site variation in evolutionary rate. Confidence in the inferred phylogenetic relationships was determined by construction of a majority-rule consensus tree based on alternative topologies not considered to be statistically significantly different from the optimal tree. The results are in agreement with earlier studies in the assignment of individual taxa to specific sequence groups. Several relationships not previously noted among sequence groups are indicated, whereas other relationships previously supported are contradicted. All plastids cluster as a strongly supported monophyletic group arising near the root of the cyanobacterial line of descent. PMID- 10461382 TI - Plastids and protein targeting. AB - Plastids with two bounding membranes--as exemplified by red algae, green algae, plants, and glaucophytes--derive from primary endosymbiosis; a process involving engulfment and retention of a cyanobacterium by a phagotrophic eukaryote. Plastids with more than two bounding membranes (such as those of euglenoids, dinoflagellates, heterokonts, haptopytes, apicomplexa, cryptomonads, and chlorarachniophytes) probably arose by secondary endosymbiosis, in which a eukaryotic alga (itself the product of primary endosymbiosis) was engulfed and retained by a phagotroph. Secondary endosymbiosis transfers photosynthetic capacity into heterotrophic lineages, has apparently occurred numerous times, and has created several major eukaryotic lineages comprising upwards of 42,600 species. Plastids acquired by secondary endosymbiosis are sometimes referred to as "second-hand." Establishment of secondary endosymbioses has involved transfer of genes from the endosymbiont nucleus to the secondary host nucleus. Limited gene transfer could initially have served to stabilise the endosymbioses, but it is clear that the transfer process has been extensive, leading in many cases to the complete disappearance of the endosymbiont nucleus. One consequence of these gene transfers is that gene products required in the plastid must be targeted into the organelle across multiple membranes: at least three for stromal proteins in euglenoids and dinoflagellates, and across five membranes in the case of thylakoid lumen proteins in plastids with four bounding membranes. Evolution of such targeting mechanisms was obviously a key step in the successful establishment of each different secondary endosymbiosis. Analysis of targeted proteins in the various organisms now suggests that a similar system is used by each group. However, rather than interpreting this similarity as evidence of an homologous origin, I believe that targeting has evolved convergently by combining and recycling existing protein trafficking mechanisms already existing in the endosymbiont and host. Indeed, by analyzing the multiple motifs in targeting sequences of some genes it is possible to infer that they originated in the plastid genome, transferred from there into the primary host nucleus, and subsequently moved into the secondary host nucleus. Thus, each step of the targeting process in "second-hand" plastids recapitulates the gene's previous intracellular transfers. PMID- 10461383 TI - The non-photosynthetic plastid in malarial parasites and other apicomplexans is derived from outside the green plastid lineage. AB - The discovery of a non-photosynthetic plastid genome in Plasmodium falciparum and other apicomplexans has provided a new drug target, but the evolutionary origin of the plastid has been muddled by the lack of characters, that typically define major plastid lineages. To clarify the ancestry of the plastid, we undertook a comprehensive analysis of all genomic characters shared by completely sequenced plastid genomes. Cladistic analysis of the pattern of plastid gene loss and gene rearrangements suggests that the apicomplexan plastid is derived from an ancestor outside of the green plastid lineage. Phylogenetic analysis of primary sequence data (DNA and amino acid characters) produces results that are generally independent of the analytical method, but similar genes (i.e., rpoB and rpoC) give similar topologies. The conflicting phylogenies in primary sequence data sets make it difficult to determine the the exact origin of the apicomplexan plastid and the apparent artifactual association of apicomplexan and euglenoid sequences suggests that DNA sequence data may be an inappropriate set of characters to address this phylogenetic question. At present we cannot reject our null hypothesis that the apicomplexan plastid is derived from a shared common ancestor between apicomplexans and dinoflagellates. During the analysis, we noticed that the Plasmodium tRNA-Met is probably tRNA-fMet and the tRNA-fMet is probably tRNA-Ile. We suggest that P. falciparum has lost the elongator type tRNA Met and that similar to metazoan mitochondria there is only one species of methionine tRNA. In P. falciparum, this has been accomplished by recruiting the fMet-type tRNA to dually function in initiation and elongation. The tRNA-Ile has an unusual stem-loop in the variable region. The insertion in this region appears to have occurred after the primary origin of the plastid and further supports the monophyletic ancestory of plastids. PMID- 10461384 TI - Voltage-dependent reversal of anodic galvanotaxis in Nyctotherus ovalis. AB - Aerobic and anaerobic ciliates swim towards the cathode when they are exposed to a constant DC field. Nyctotherus ovalis from the intestinal tract of cockroaches exhibits a different galvanotactic response: at low strength of the DC field the ciliates orient towards the anode whereas DC fields above 2-4 V/cm cause cathodic swimming. This reversal of the galvanotactic response is not due to backward swimming. Rather the ciliates turn around and orient to the cathode with their anterior pole. Exposure to various cations, chelators, and Ca(2+)-channel inhibitors suggests that Ca(2+)-channels similar to the "long lasting" Ca(2+) channels of vertebrates are involved in the voltage-dependent anodic galvanotaxis. Evidence is presented that host-dependent epigenetic factors can influence the voltage-threshold for the switch from anodic to cathodic swimming. PMID- 10461385 TI - Fractionation of sporogonial stages of the microsporidian Encephalitozoon cuniculi by Percoll gradients. AB - Microsporidia are obligate intracellular parasites that are increasingly recognized as a cause of opportunistic infections in immunocompromised individuals. Encephalitozoon cuniculi has been identified in humans with AIDS and infects a wide range of mammalian hosts. Little is known about the metabolic processes that regulate growth and replication of microsporidia. Examination of the individual stages of development will facilitate such studies and reveal possible targets for drug therapy. The purpose of this study was to fractionate and purify stages of the microsporidian life cycle. Encephalitozoon cuniculi were cultured in RK-13 cells. The tissue supernatants containing multiple parasite stages, empty microsporidial husks and host cell debris were collected, washed, and subjected to differential centrifugation in 80% stock isotonic Percoll. Transmission electron microscopy and SDS-polyacrylamide gel electrophoresis were used to compare the content and purity of each fraction. Mature spores formed a band at a density of approximately 1.138 g/ml. Sporoblasts were found at densities between 1.102 g/ml and 1.119 g/ml. A mixture of sporonts, sporoblasts, microsporidial husks, and cell debris remained at the top of the gradient and additional centrifugation in 30% and 50% Percoll resulted in separation of these stages. These results represent the first step toward fractionating stages of microsporidia infecting humans. PMID- 10461386 TI - Characterization of two Perkinsus spp. from the softshell clam, Mya arenaria using the small subunit ribosomal RNA gene. AB - Sequence analysis and riboprinting of the small subunit ribosomal RNA genes were used to characterize two morphologically different Perkinsus species isolates from the gill (G117) and the hemolymph (H49) of the softshell clam, Mya arenaria. Sequence data of the polymerase chain reaction amplified ribosomal RNA loci of G117 and H49 indicated that these genes are 1803 and 1806 base-pair long, respectively. A sequence similarity of > 98.9% was calculated among ribosomal RNA sequences of the two isolates of this study and the published sequences of Perkinsus marinus from the American eastern oyster, Crassostrea virginica, and Perkinsus sp. from the blood cockle of the Australian mollusc, Anadara trapezia. From a phylogenetic tree obtained from Jukes-Cantor distances of the aligned ribosomal RNA gene sequences of 13 eukaryotic taxa using the Neighbor-Joining method, we showed that G117 and H49 clustered within the genus Perkinsus. Guided by the sequence data of Perkinsus marinus (accession # X75762) and Perkinsus sp. (accession # L07375), restriction endonucleases were selected for restriction fragment analysis of polymerase chain reaction products of the small subunit ribosomal RNA genes (riboprinting). Riboprinting was used to distinguish the four members of the genus Perkinsus from each other. PMID- 10461388 TI - Keto-acid oxidoreductases in the anaerobic protozoa. AB - In anaerobes, decarboxylation of pyruvate is executed by the enzyme pyruvate:ferredoxin oxidoreductase, which donates electrons to ferredoxin. The pyruvate:ferredoxin oxidoreductase and its homologues utilise many alternative substrates in bacterial anaerobes. The pyruvate:ferredoxin oxidoreductase from anaerobic protozoa, such as Giardia duodenalis, Trichomonas vaginalis, and Entamoeba histolytica have retained this diversity in usage of alternative keto acids for energy production utilising a wide variety of substrates. In addition to this flexibility, both T. vaginalis and G. duodenalis have alternative enzymes that are active in metronidazole-resistant parasites and that do not necessarily involve donation of electrons to characterized ferredoxins. Giardia duodenalis has two oxoacid oxidoreductases, including pyruvate:ferredoxin oxidoreductase and T. vaginalis has at least three. These alternative oxoacid oxidoreductases apparently do not share homology with the characterized pyruvate:ferredoxin oxidoreductase in either organism. Independently, both G. duodenalis and T. vaginalis have retained alternative oxoacid oxidoreductase activities that are clearly important for the survival of these parasitic protists. PMID- 10461387 TI - Analysis of intraspecific sequence variation among eight isolates of the rumen symbiont, Isotricha prostoma (Ciliophora), from two continents. AB - The internally transcribed spacer regions 1 and 2 (ITS-1 and ITS-2) and the 5.8S ribosomal RNA gene of Isotricha prostoma were examined for intraspecific sequence variation. There were no differences in the ITS-1/5.8S/ITS-2 region among cattle and sheep isolates of I. prostoma from Australia, Canada, and the United States, indicating that this region is 100% conserved among eight isolates from two continents. PMID- 10461389 TI - Spermatozoa, DNA binding and transgenic animals. AB - The idea that sperm cells could be used as an effective tool for introducing exogenous DNA into an oocyte at fertilization is generally regarded with scepticism. However, in recent years, several investigators have been working on different aspects of this intriguing research topic. In the present review, their results are summarised and discussed. Sections have been dedicated to the way DNA molecules bind to spermatozoa of different species, to the events regulating such binding, to the fate of the DNA within sperm cells, and to the attempts made to produce transgenic animals with this method. The data available on the interaction between DNA and spermatozoa begin to explain how this event takes place and how it is regulated. However, the stable integration of exogenous genes into the genome of adult animals mediated by sperm cells is a very rare event, although several reports describe forms of partial success. Available evidence suggests that changes to the DNA molecules, occurring mostly within the oocyte, represents the limiting step in the production of transgenic animals using spermatozoa as vectors of exogenous genes. At present there are not enough data to understand what happens to sperm-associated DNA upon its entrance into the oocyte at fertilization. Therefore, it has not yet been resolved whether sperm mediated gene transfer is a possible way to manipulate the genome or if evolution has imposed some unsurpassible barriers to its use. PMID- 10461390 TI - Biosafety of E. coli beta-glucuronidase (GUS) in plants. AB - The beta-glucuronidase (GUS) gene is to date the most frequently used reporter gene in plants. Marketing of crops containing this gene requires prior evaluation of their biosafety. To aid such evaluations of the GUS gene, irrespective of the plant into which the gene has been introduced, the ecological and toxicological aspects of the gene and gene product have been examined. GUS activity is found in many bacterial species, is common in all tissues of vertebrates and is also present in organisms of various invertebrate taxa. The transgenic GUS originates from the enterobacterial species Escherichia coli that is widespread in the vertebrate intestine, and in soil and water ecosystems. Any GUS activity added to the ecosystem through genetically modified plants will be of no or minor influence. Selective advantages to genetically modified plants that posses and express the E. coli GUS transgene are unlikely. No increase of weediness of E. coli GUS expressing crop plants, or wild relatives that might have received the transgene through outcrossing, is expected. Since E. coli GUS naturally occurs ubiquitously in the digestive tract of consumers, its presence in food and feed from genetically modified plants is unlikely to cause any harm. E. coli GUS in genetically modified plants and their products can be regarded as safe for the environment and consumers. PMID- 10461391 TI - Tissue-specific expression of human salivary mucin gene, MUC7, in transgenic mice. AB - The MUC7 gene encodes the protein core of the low molecular weight human salivary mucin (MG2, mucin glycoprotein 2) and is expressed in a tissue-specific manner in salivary glands. The purpose of this study was to examine MUC7 expression by transgenic mouse technology. A 16 kb DNA fragment, containing the MUC7 gene (10 kb) and 3 kb of the upstream and 3 kb of the downstream sequences, was used to generate transgenic mice. We have identified five transgenic founder mice which were propagated as individual transgenic lines and analysed. Tissues of transgenic offspring from each line were analysed by RT-PCR to determine the sites of the MUC7 expression. The results indicated that only line 3 and line 5 expressed the MUC7 gene in salivary glands. The level of MUC7 expression in selected tissues was then determined by northern blot analyses. The results showed that line 3 mice contained high levels of MUC7 transcripts in the sublingual glands of both males and females and indicated low levels of MUC7 transcripts in the submandibular glands of females. No MUC7 expression was detected in this line by northern blot analysis in any other tissue tested. On the other hand, no expression of MUC7 was detected in any tissues of line 5 mice examined by northern blot analysis. A Southern blot analysis of human and mouse genomic DNA demonstrated multiple copies of the MUC7 transgene in line 3 and a single copy in line 5. Collectively, these results indicate that the regulatory sequences required for the tissue-specific expression of MUC7 are within the MUC7 transgene. However, the sequences necessary for expression comparable to that of MUC7 in human salivary glands may be missing from this construct. Western blot analysis of protein extracts from different tissues of transgenic mice line 3 showed that MUC7 gene product was produced in the submandibular-sublingual gland complex of both male and female mice and not in the other tissues examined. PMID- 10461392 TI - Rescue of an MMTV transgene by co-integration reveals novel locus control properties of the ovine beta-lactoglobulin gene that confer locus commitment to heterogeneous tissues. AB - In an attempt to enhance the frequency and level of expression of a poor performing MMTV-driven transgene, we co-integrated this construct with the ovine beta-lactoglobulin (BLG) gene in transgenic mice. Seven lines of transgenic mice possessing co-integrated BLG and MMTV-RZ5 transgenes were compared with 12 lines of mice that possessed only the MMTV-RZ5 construct. Co-integration enhanced the frequency of expression in the mammary gland from two out of 12 lines for the MMTV-RZ5 transgene alone, to five out of seven when co-integrated with BLG. Surprisingly, co-integration also resulted in co-expression of the two transgenes in the salivary gland, lung and spleen in addition to the mammary gland. Furthermore, both transgenes were expressed in virgin animals, and throughout pregnancy and lactation, suggesting that the developmental regulation of the locus follows that of the MMTV-promoter. These findings represent a novel locus control property of the ovine BLG gene that confers commitment of the locus to the mammary gland, but also to a range of heterogeneous tissues possibly defined by the second promoter at the locus. PMID- 10461394 TI - Gallium replication in aquatic and nonaquatic organism scanning electron microscopy. AB - A technique using pure gallium metal as a replication material is reported for biological surface scanning electron microscopy (SEM). The technique first directly enables aquatic organisms in water to be replicated due to gallium's low melting point and, second, reproduces surface structures and images of the two dimensional (2D) distribution of substances transferred from the original surface to the gallium surface due to gallium's high surface tension. An aquatic protozoan in water was directly replicated to show its typical surface structures. The technique was then used to visualize human hair surface structures and 2D transferred substance distribution using X-ray microanalysis. PMID- 10461393 TI - Double pronuclei injection of DNA into zygotes increases yields of transgenic mouse lines. AB - Transgenic mice are increasingly used for gene function and regulation studies of mammalian genes. A major limitation is the necessity to produce a large number of founder animals to obtain one line with the desired expression pattern. We developed a method, the 'double pronuclei injection', that doubles the yield of transgenic mouse lines obtained from each injection session, thereby reducing the time, effort and costs of generating transgenic mice. Three transgenic vectors were microinjected into the male and female pronuclei of zygotes. Approximately half of the resulting born mice were transgenic. This represented a 60% increase in the yield of founders per injected zygote, and a 100% increase in the yield of transgenic mice per born animal, when compared to yields obtained using single pronucleus injection. This method should prove useful for generating large numbers of transgenic mice for gene regulation studies and for conditional gene ablation. PMID- 10461395 TI - Health hazard evaluation of methyl bromide soil fumigations. PMID- 10461396 TI - New draft OSHA ergonomics rule. PMID- 10461397 TI - Industrial hygiene databases: structure and function. PMID- 10461398 TI - Control of exposure to perchloroethylene in commercial drycleaning (substitution). National Institute for Occupational Safety and Health. PMID- 10461399 TI - The OSHA poultry initiative. PMID- 10461400 TI - Measurements of oxygen concentration of air within totally encapsulating chemical protective suits. AB - This study measured oxygen concentrations of air within totally encapsulating chemical-protective (TECP) suits during simulated hazardous materials emergency response operations. This was done to allow assessment of the suitability of use of air within TECP suits as an emergency air supply during escapes from contaminated areas following loss of primary air supply. Samples of air were extracted through a probe installed in TECP suits while the suits were worn during simulated hazardous materials control operations. Data on oxygen concentration were obtained using direct reading equipment and through Orsat analysis of samples. Results indicate that the air within a TECP suit becomes significantly oxygen-deficient soon after the suit is closed, but generally maintains oxygen concentrations somewhat above those known to produce significant physiological effects. However, due to the further reduction of oxygen content inherent in breathing the air within TECP suits, it is anticipated that symptoms of oxygen deficiency sufficient to impair escape could be experienced in some instances. Recommendations are made for modifying current training practices to better prepare personnel working in Level A chemical-protective ensembles to cope with loss of air supply. PMID- 10461401 TI - An Australian study to evaluate worker exposure to chrysotile in the automotive service industry. AB - A study was conducted in Sydney, Australia, in 1996 to investigate the current exposure levels, control technologies, and work practices in five service garages (four car and one bus), three brake bonding workshops, and one gasket processing workshop. This study formed part of the assessment of chrysotile as a priority existing chemical under the Australian National Industrial Chemicals Notification and Assessment Scheme. A total of 68 (11 personal and 57 area) air samples were collected, in accordance with the Australian standard membrane filter method. Fiber concentrations were determined by the traditional phase contrast microscopy (PCM) method and 16 selected samples were analyzed by the more powerful transmission electron microscopy (TEM). Chrysotile exposure of car mechanics measured by PCM was typically below the reportable detection limit of 0.05 f/mL, irrespective of whether disc brake, drum brake, or clutch was being serviced. These low levels can be attributed to the wet cleaning or aerosol spray methods used in recent years to replace the traditional compressed air jet cleaning. In the three brake shoe relining workshops, task-specific exposure reached up to 0.16 f/mL in the processes of cutting and radius grinding. TEM results were generally higher, due to its higher resolution power. The median diameter on samples taken from the service garages (passenger cars), as determined by TEM, was 0.5-1.0 micron; and was between 0.2-0.5 micron for the brake bonding and gasket processing workshops, while that for the bus service depot was 0.1-0.2 micron. Most of the respirable fibers (84%, mainly forsterite) from the bus service depot were below 0.2 micron in diameter which is the resolution limit of PCM. In the brake bonding and gasket cutting workshops, 34 percent and 44 percent of the chrysotile fibers were below 0.2 micron in diameter. PMID- 10461402 TI - Surveillance of construction worker injuries: the utility of trade-specific analysis. AB - Construction is a dangerous industry, with high rates of both fatal and nonfatal injuries. To learn more about the causes of nonfatal construction worker injuries, and to identify injury cases for further work site-based investigations or prevention programs, we established an emergency department-based surveillance program in November 1990. This article describes circumstances of injury, diagnoses, and demographic characteristics of injured construction workers for 2,791 cases identified through mid-August, 1997. Lacerations and strains and sprains were the most frequent diagnoses; cutting and piercing objects were the leading causes of injury among all construction workers, followed by falls and overexertion. Because of the variety of work performed in this industry, more detailed injury descriptions, by trade, are most useful for thinking about injury prevention. To illustrate this, we profile injury patterns among workers from four specific trades: carpenters, electricians, plumbers, and ironworkers. Areas of concern highlighted by the trade-specific analyses include eye injuries among plumbers; falls from ladders among electricians and plumbers; slips, trips, and falls on the same level among ironworkers; electrical exposure among electricians; and, amputations among carpenters. PMID- 10461403 TI - A description of factors affecting hazardous waste workers' use of respiratory protective equipment. AB - This article describes the first phase of a study that was designed to gain an understanding of hazardous waste workers' attitudes and beliefs about the use of respiratory protective equipment. Exploratory, open-ended interviews were conducted among 28 respirator users at a US Department of Energy facility. Subjects were asked to describe their knowledge, attitudes, and beliefs about their risks to hazards at their worksites and to discuss their use of respiratory protective equipment. A detailed content analysis of the interviews resulted in the generation of a taxonomy of issues and concerns which fell into three general categories: 1) Knowledge, Beliefs, and Attitudes, 2) Physical and Psychological Effects, and 3) External Influences. Knowledge, Beliefs, and Attitudes included Training, Fit Testing, Medical Clearance, Work Exposures, Respirator Use, and Vulnerability to Disease. Physical and Psychological Effects included Somatic/Health Effects, Personal Comfort, Visual Effects, Fatigue, Communication, and Anxiety. External Influences included Structural Environment, Quality and Availability of Equipment, Other PPEs, Co-Worker Influence, Supervisor Influence, and Organizational Culture. The findings from this study have important implications to training and education programs. Effective respiratory protection programs depend on a knowledge of the factors that affect workers' use of equipment. This study suggests that efforts to assure equipment comfort and fit, to assist workers who see and hear less well as a result of their equipment, and to develop strategies to allay worker anxiety when wearing equipment should all be components of a program. An organizational culture that supports and abets the appropriate use of equipment is also a critical element in a successful program. PMID- 10461404 TI - Surface composition of silica particles embedded in an Australian bituminous coal. AB - The composition and structure of the surface layers of a series of silica particles (10-20 microns across), embedded in a bituminous coal from the Whybrow seam, Sydney Basin, Australia, have been characterized in situ using time-of flight secondary ion mass spectrometry (TOFSIMS), ion imaging, and depth profiling. The silica particles investigated are typically encased in a multilayered shell, the composition of which differs from average composition of both the silica and the bulk coal. The analysis directly demonstrates the presence of a silanol-rich (Si-OH) interfacial layer 3 nm in thickness. This silanol-rich region separates the bulk silica and a complex non-silica layer encasing the particles. The interfacial region also shows significant lithium enrichment (approximately fivefold over bulk) which implies diffusion and precipitation of lithium-containing species during the authigenetic formation of the surface layers of the silica grains. The outer layer encasing the silica particles is 10 nm in thickness and is composed of clays and carbonates, and, in some cases, includes organic material. The elemental constituents of this layer include aluminium, sodium, potassium, magnesium, iron, and lesser amounts of titanium and copper. The variation in the aluminium concentration from the outermost surface to the deeper layers is less than that of other non-silica species. A relatively high amount of calcium is found associated with the silica bulk. Although only non-respirable-sized silica particles are examined in this work, the methods of analysis developed have potential in providing an insight into the surface composition of respirable particles and in further studies of the surface bioavailability of silica species. PMID- 10461405 TI - The indigenous Sea Gypsy divers of Thailand's west coast: measurement of carbon monoxide in the breathing air. AB - Approximately 400 indigenous divers live and work on Thailand's west coast. They dive with surface supplied air from primitive compressor units mounted on open boats which measure from seven to 11 meters in length. It was suspected that carbon monoxide was present in the breathing air of at least the gasoline-driven compressor units. To determine the presence of carbon monoxide gas in the breathing air, compressed air from the compressor was pumped through the diver air supply hose through a plenum (monitoring) chamber established on the boat. After a compressor warm-up of 15 minutes, the diving air was measured with the boat at eight different bearings to the wind, each 45 degrees apart at intervals of five minutes. Three of the four gasoline-driven compressor units tested showed presence of carbon monoxide in the breathing air. One diesel-driven unit showed a very low concentration of carbon monoxide (3-4 ppm) and six diesel-driven units showed no detectable carbon monoxide. Although not tested, diesel exhaust emissions could also enter the breathing air by the same route. A locally made modification to the compressor air intake was designed and successfully tested on one gasoline-driven compressor unit. An information sheet on the hazards of carbon monoxide as well as on the modification has been developed for distribution among the villages. PMID- 10461406 TI - The predictive value of lymphoscintigraphy for nodal metastases of cutaneous melanoma. AB - Lymphoscintigraphy is used to identify ambiguous drainage patterns in cutaneous melanoma of the head, neck and trunk. This study evaluated the efficacy and reliability of lymphoscintigraphy to identify the drainage nodes in 51 patients undergoing both elective and therapeutic lymphadenectomy over a seven-year period. All 13 patients who had lymph node metastases during this follow-up period had the metastatic disease in the very lymph node basins identified by lymphoscintigraphy. Most significantly, none of the 51 patients had metastatic disease in lymphatic basins that were not previously identified by lymphoscintigraphy. PMID- 10461407 TI - A logical approach to the injection treatment of varicose and "spider" veins. AB - The incurable nature of varicose veins mandates a re-evaluation of excisional surgery both as appropriate therapy and as the gold standard against which to judge alternative treatments. An injection techniques has been developed which enables synchronous in situ obliteration of all varicose veins and superficial venous ectasia, regardless of their size or extent, and eliminates the need to identify incompetent perforators. The results of 742 patients (1,425 legs) were evaluated retrospectively from two to 12 years following initial therapy. All patients experienced total relief of painful symptoms and 95% were very pleased with the final appearance. Some residual brown pigmentation or a scar from an injection ulcer were the reasons for less than total satisfaction in the remaining 5%. None regretted having undergone the procedure. Because of the difficulty in sustaining compression in the inguinal area, 6% of patients required a concomitant sapheno-femoral disconnection under local anesthesia immediately prior to injection therapy, but none required stripping. The results repudiate the generally held belief that injection therapy is only of value for treating postsurgical recurrences, or for eliminating small varicosities and superficial capillary ectasia (spider veins) for cosmetic improvement. PMID- 10461408 TI - Iatrogenic vitamin D intoxication: report of a case and review of vitamin D physiology. PMID- 10461409 TI - Sympathetic schwannoma: a case report. AB - Schwannomas (also known as Neurilemomas or Neurinomas) are benign tumors of the nervous system that originate in the neural sheath and most commonly occur as solitary encapsulated subcutaneous tumors in otherwise healthy individuals. When they present as multiple tumors, they are histologically indistinguishable from the solitary tumors and, in such a case, they may be associated with neurofibromatosis 2 (NF2) in which the sensory roots of the cranial and spinal nerves are affected more often than the motor roots and the hallmark tumor of which is a vestibular schwannoma (also known as acoustic neuroma). They also may exist as a distinct clinical entity without any stigmata of neurofibromatosis known as schwannomatosis. Generally, schwannomas are less than 5 cm in diameter but sizes greater than 14 cm have been reported. They present as slow growing masses and may present with pain or neurological symptoms. Here we report a patient with sympathetic schwannoma who presented with right flank pain. PMID- 10461410 TI - An acute sign of ischemic infarction: the hyperdense middle cerebral artery. AB - Ischemic cerebrovascular accidents are a leading cause of death with significant disability common among the survivors. The hyperdense artery sign seen on computed tomography is a useful finding in the early recognition of nonhemorrhagic cerebral infarction. We report a case of a hyperdense middle cerebral artery. PMID- 10461411 TI - Infliximab (Remicade). PMID- 10461412 TI - Improving the care of patients with community-acquired pneumonia: a multihospital collaborative QI project. AB - BACKGROUND: Performance of several processes of care was measured in eight acute care hospitals in Connecticut which provided inpatient treatment to 713 elderly patients with community-acquired pneumonia (CAP). BASELINE DATA ABSTRACTION AND FEEDBACK: Chart review feedback was provided, and the hospitals were requested to design their own quality improvement (QI) interventions, after which re examination of process of care performance was conducted. HOSPITAL QI INTERVENTIONS: Six of the eight hospitals had submitted QI plans. The quality indicators dealing with timeliness of antibiotic delivery were specifically addressed by five hospitals. However, each hospital also picked one or two other process of care for intervention. RESULTS: The mean time to antibiotic administration decreased from 5.5 hours (+/- 0.2) to 4.7 hours (+/- 0.3; P < 0.0001), and the percentage of patients who received antibiotics within four hours increased from 41.5% to 61.6% (P < 0.0001). DISCUSSION: This project called for obtaining buy-in from both the clinician and administrative representatives of each hospital early in the process. In this way, the targeted processes of care were likely to have relevance for each of the participating hospitals. Education of practicing physicians and other health professionals, as the method chosen by each hospital to address delays in antibiotic administration, appears to have been successful in this project as part of a multifaceted intervention. The project also helped establish a collegial environment that has served as the basis for more ambitious pneumonia QI projects. SUMMARY AND CONCLUSIONS: Widespread improvements in process of care performance can result from hospitals' participation in a Quality improvement Organization collaboration. PMID- 10461413 TI - Leaders in medicine. Donald L. Cooper, MD. PMID- 10461414 TI - 54-year-old man with progressive dementia: a clinicopathologic correlation conference from the University of Oklahoma College of Medicine. PMID- 10461415 TI - Hepato-biliary abnormalities secondary to ceftriaxone use: a case report. AB - Ceftriaxone was approved in 1997 for the treatment of otitis media despite previous studies that documented an association of ceftriaxone with elevated hepato-biliary enzymes and transient biliary stasis. The case cited here highlights the need for continued awareness education for physicians who may use ceftriaxone to treat common illnesses such as acute exudative tonsillitis and otitis media in children. Specifically, for children with a family history of gallbladder, biliary tract, liver or pancreas dysfunction, ceftriaxone may not be the drug of choice since the likelihood of complications is increased in this population. Additionally, ceftriaxone may cause problems in either adults or children with preexisting disease, who may not be well-nourished, or who may be dehydrated. PMID- 10461416 TI - Information management in medicine. PMID- 10461417 TI - Dr. Kelly West and a brief history of the diabetes epidemic of American Indians. PMID- 10461418 TI - Choledochal cyst associated the with anomalous union of pancreaticobiliary duct (AUPBD) has a more grave clinical course than choledochal cyst alone. AB - OBJECTIVE: Since choledochal cyst is frequently associated with the anomalous union of pancreaticobiliary duct (AUPBD), AUPBD has been regarded to be the etiologic factor of choledochal cyst. However, the clinical significance of AUPBD an patients with choledochal cyst has not been clearly defined. Therefore, to clarify the significance of AUPBD in choledochal cyst patients, we compared the clinical features of patients with choledochal cyst according to the presence or absence of AUPBD. METHODS: Among 52 cases which were diagnosed as choledochal cyst out of 5,037 ERCP referrals between August 1990 and December 1996, we selected 44 cases, in which the pancreaticobiliary junction was clearly visualized on cholangio-pancreaticography. These cases were divided into AUPBD present group (n = 28) and AUPBD-absent group (n = 16). Clinical features were compared between the two groups. Furthermore, in AUPBD-present group, clinical data were also analyzed according to Kimura's classification of AUPBD. RESULTS: In our study, AUPBD was associated with choledochal cyst in 28 (64%) cases. AUPBD was found only in type I and IV according to Todani's classification of choledochal cyst. There were no significant differences between the AUPBD-present group and the AUPBD-absent group in the incidence of gallstone disease, while the incidence of acute inflammation was 93% (26/28) in the AUPBD-absent group (p < 0.01). Carcinoma developed only in the AUOBD-present group (9/28, 32%) (p < 0.05). Pancreatic disorders (i.e. pancreatic stone, pancreatitis or pancreatic cancer) occurred in 12 of 28 cases in the AUPBD-present group (43%), while only in 1 of 16 cases in the AUPBD-absent group (6%) (p < 0.05). CONCLUSION: AUPBD associated with choledochal cyst may have implications not only as a possible etiologic factor but also as an important factor that may affect the clinical course, surgical planning and prognosis. In cases with choledochal cyst, we should make an effort to evaluate the presence of AUPBD. PMID- 10461419 TI - Effect of eradication of Helicobacter pylori on the benign gastric ulcer recurrence--a 24 month follow-up study. AB - OBJECTIVES: To evaluate the effect of eradication of Helicobacter pylori (H. pylori) on the recurrence of benign gastric ulcer (BGU) in the patients with BGU. METHODS: This study was performed for 40 H. pylori-positive BGU patients cured of BGU and H. pylori eradicated, and for 25 H. pylori-positive patients (non eradicated group) who were not treated with H. pylori eradication regimen or H. pylori was not eradicated. Four different methods--CLOtest, microscopy of Gram stained mucosal smear, culture and histology of modified Giemsa staining--were taken for identifying colonization of H. pylori before treatment, and 4 weeks after completion of triple therapy. For the control group in which triple therapy was not tried, follow-up gastroscopy was done to confirm the healing of the ulcer. To detect BGU recurrence, the gastroscopy was performed at 6, 12, 18, and 24 months after therapy. RESULTS: In the non-eradicated group, the BGU recurrence rate was 16% within 6 months, 40% within 1 year, 56% within 18 months and 60% within 2 years. The respective recurrence rates in the 40 patients in whom the bacteria had been eradicated were 0%, 7.5%, 10% and 10% (4 patients), respectively. Among the four BGU-recurred patients in whom H. pylori had been eradicated, one patient was found to have BGU recurring with H. pylori positive again in one year, and another two patients had NSAIDs ingestion history. CONCLUSION: The eradication of H. pylori in patients with BGU reduces the recurrence of BGU. In addition, the major causes of BGU recurrence look like NSAIDs ingestion and reinfection of H. pylori. PMID- 10461420 TI - Imipenem-cilastatin versus sulbactam-cefoperazone plus amikacin in the initial treatment of febrile neutropenic cancer patients. AB - The treatment of infectious complications in cancer patients has evolved as a consequence of the developments in the chemotherapy of cancer patients. In this prospective, randomized study, we compared imipenem-cilastatin and sulbactam cefoperazone with amikacin in the empiric therapy of febrile neutropenic (< 1000/mm3) patients with liquids and solid tumours. Of 30 evaluable episodes, 15 were treated with imipenem-cilastatin and 15 were treated with sulbactam cefoperazone plus amikacin. 73% of episodes were culture-positive: gram-positive pathogens accounted for 62% of the isolates. Bacteremia was the most frequent site of infection. The initial clinical response rate for both regimens was 60% (p > 0.05). No major adverse effects occurred. This study demonstrated that imipenem-cilastatin monotherapy and combination therapy of sulbactam-cefoperazone plus amikacin were equally effective empiric therapy for febrile granulocytopenic cancer patients. PMID- 10461421 TI - Transesophageal echocardiographic detection of thoracic aortic plaque could noninvasively predict significant obstructive coronary artery disease. AB - OBJECTIVE: Previous pathologic and roentgenographic studies have suggested a relation between aortic plaque and coronary artery disease but have lacked clinical utility. The study was undertaken to elucidate whether atherosclerotic aortic plaque detected by transesophageal echocardiography can be a clinically useful marker for significant obstructive coronary artery disease. METHODS: Clinical and angiographic features and intraoperative transesophageal echocardiographic findings were prospectively analyzed in 131 consecutive patients (58 women and 73 men, aged 17 to 75 years [mean 54 +/- 12]) undergoing open heart surgery. Significant obstructive coronary artery disease was defined as > or = 50% stenosis of > or = 1 major branch. RESULTS: Seventy-six (58%) of 131 patients were found to have obstructive coronary artery disease. In 76 patients with significant coronary artery disease, 71 had thoracic aortic plaque. In contrast, aortic plaque existed in only 10 of the remaining 55 patients with normal or minimally abnormal coronary arteries. The presence of aortic plaque on transesophageal echocardiographic studies had a sensitivity of 93%, a specificity of 82% and positive and negative predictive values of 88% and 90%, respectively, for significant coronary artery disease. There was a significant relationship between the degree of aortic intimal changes and the severity of coronary artery disease (r = 0.74, P < 0.0001). Multivariate logistic regression analysis of patient age, sex, risk factors of cardiovascular disease and transesophageal, echocardiographic findings revealed that atherosclerotic aortic plaque was the most significant independent predictor of coronary artery disease. CONCLUSION: This study indicates that transesophageal echocardiographic detection of atherosclerotic plaque in the thoracic aorta is useful in the noninvasive prediction of the presence and severity of coronary artery disease. PMID- 10461422 TI - Protective effect of chlormethiazole, a sedative, against acetaminophen-induced liver injury in mice. AB - OBJECTIVES: The hepatotoxicity of acetaminophen is not a result of the parent compound but is mediated by its reactive metabolite N-acetyl-p-benzoquinone imine. Cytochrome P4502E1 (CYP2E1) is the principal enzyme of this biotransformation, which accounts for approximately 52% of the bioactivation in human microsomes. Recently, chlormethiazole a sedative drug, is reported to be an efficient inhibitor of CYP2E1 activity in human beings. In this study we wished to evaluate whether chlormethiazole, an inhibitor of CYP2E1, could prevent acetaminophen-induced liver injury in mice. METHODS: Acetaminophen, at doses ranging from 200 to 600 mg/kg, was injected into the peritoneum of female C57BL/6 inbred mice fasted for four hours. Chlormethiazole (60 mg/kg) or 5% dextrose water was given 30 min before or 2 h after acetaminophen. Serum aminotransferase activities, histologic index score, survival rate and hepatic malondialdehyde levels were compared. RESULTS: Pretreatment with chlormethiazole 30 min before 400 mg/kg of acetaminophen completely inhibited acetaminophen-induced liver injury (median 118.5 U/L, range 75 to 142 vs. 14,070 U/L, range 5980 to 27,680 for AST; 49 U/L, range 41 to 64 vs. 15,330 U/L, range 13,920 to 15,940 for ALT). In mice receiving chlormethiazole 2 h after acetaminophen, the mean AST and ALT levels were also less elevated, reaching only 20% of the value of acetaminophen only group. These protective effects were confirmed histologically. Whereas more than 50% of mice died at 500 mg/kg of acetaminophen, all the mice pretreated with chlormethiazole survived at the same dose. CONCLUSION: Chlormethiazole effectively reduces acetaminophen-induced liver injury in mice. Further studies are needed to assess its role in humans. PMID- 10461423 TI - Effect of cilostazol on the neuropathies of streptozotocin-induced diabetic rats. AB - OBJECTIVES: This study examined the effect of cilostazol, a potent phosphodiesterase inhibitor, on the progression of neuropathies associated with streptozotocin-induced diabetes mellitus in Sprague-Dawley rats. METHODS: Eight weeks after streptozotocin treatment, a pelleted diet containing 0.03% cilostazol (15 mg/kg body weight) was given for four weeks. Body weight, blood glucose level, motor nerve conduction velocity (MNCV), myelinated fiber density and size distribution of sciatic nerves were compared between age-matched normal rats (Group 1), control diabetic rats (Group 2) and cilostazol-treated diabetic rats (Group 3). RESULTS: Body weight was significantly reduced and blood glucose level was significantly increased in diabetic rats (Group 2 and 3) compared to normal rats. MNCV and cAMP content of sciatic nerves were significantly reduced in diabetic rats 12 weeks after streptozotocin treatment. Myelinated fiber size and density were also significantly reduced, and thickening of the capillary walls and duplication of the basement membranes of the endoneural vessels were observed in the diabetic rats. Whereas both body weight and blood glucose level of Group 3 did not differ significantly from those of Group 2, cilostazol treatment significantly increased MNCV and cAMP content of sciatic nerves in Group 3 but not to the levels observed in Group 1. MNCV positively correlated with cAMP content of sciatic nerves (r = 0.86; p < 0.001). Cilostazol treatment not only restored myelinated fiber density and size distribution but reversed some of the vascular abnormalities. CONCLUSION: These findings suggest that a reduced cAMP content in motor nerves may be involved in the development of diabetic neuropathy, and that cilostazol may prevent the progression of diabetic neuropathy by restoring functional impairment and morphological changes of peripheral nerves. PMID- 10461424 TI - Nasal angiocentric lymphoma with hemophagocytic syndrome. AB - OBJECTIVES: Hemophagocytic syndrome (HS) is a fatal complication of nasal angiocentric lymphoma (AL) and difficult to distinguish from malignant histiocyosis. Epstein-Barr virus (EBV)-associated HS is frequently observed in lymphoma of T-cell lineage and EBV is highly associated with nasal AL. Clinicopathologic features of 10 nasal ALs with HS were reviewed to determine the clinical significance and the pathogenetic association with EBV. METHODS: Ten patients of HS were identified from a retrospective analysis of 42 nasal ALs diagnosed from 1987 to 1996. Immunohistochemical study and in situ hybridization were performed on the paraffin-embedded tumor specimens obtained from 10 patients. Serologic study of EBV-Ab was performed in 3 available patients. RESULTS: Five patients had HS as initial manifestation, 3 at the time of relapse and 2 during the clinical remission of AL. Four patients were treated by combination chemotherapy (CHOP) and others had only supportive care. The median survival of all patients with HS was 4.1 months (range 2 days-36.5 months) and all had fatal outcome regardless of the treatment-modality. All cases were positive for UCHL1 (CD45RO) and EBV by EBER in situ hybridization. The data of serologic tests indicated the active EBV infection. CONCLUSIONS: HS is a fatal complication of nasal AL and has a high association with EBV. Reactivation of EBV may contribute to HS and further investigation of predictive factors and effective treatment of HS should be pursued in the future. PMID- 10461425 TI - Expression of ICAM-1 on the Hantaan virus-infected human umbilical vein endothelial cells. AB - OBJECTIVES: In HFRS, there is a varying degree of disseminated intravascular coagulation which was evident in the early phase of the illness. It is believed also that DIC would be the consequence, at least in part, of functional changes of endothelium resulting in kinin activation and clinical syndrome. This study investigated the role of adhesion molecule in the pathogenesis of Hantaan virus related disease. METHODS: The expression of ICAM-1 antigen on the cell membrane of human umbilical vein endothelial cells was assessed by immunohistochemistry, and ICAM-1 mRNA in the endothelial cells was assessed by in situ hybridization after Hantaan virus infection (2.6 x 10(4) PFU/mL) with the time course. RESULTS: In immunohistochemistry, the number of ICAM-1 positive cells increased with time during the 12 or 24 hours after infection. 5 to 10% of HUVECs had been positive after 12-24 hours and the number of positive cells decreased abruptly after 24 hours. Hantaan antigen had been noticed after 12 hours focally on the HUVECs but continued to proliferate into day 7 post-infection when most of HUVECs were infected by Hantaan virus. In situ hybridization showed identical patterns of ICAM-1 mRNA expression after Hantaan virus infection. CONCLUSION: It implies that the Hantaan virus infection on HUVECs would express more ICAM-1 on their surface and implicated in the pathogenesis of early clinical syndrome of HFRS. PMID- 10461426 TI - Establishment of BALB/c mice model infected with Helicobacter pylori. AB - OBJECTIVES: Considering the geographic differences in the prevalence of virulence factors such as CagA or VacA of H. pylori isolated from Korean adults compared with those from western countries, the establishment of a mouse model infected with H. pylori isolated from Korean adults is needed to investigate the pathogenesis and to develop vaccines against H. pylori infection in Korea. The aim of this study was to establish the BALB/c mouse model infected with H. pylori isolated from Korean. METHODS: Six-week-old BALB/c mice were inoculated intragastrically with 10(9) CFU of H. pylori. Loss of glandular architecture, erosions and infiltration of inflammatory cells within the lamina propria compared with normal gastric mucosa were scrutinized. Evidence for H. pylori infection was assessed by rapid urease test of gastric mucosa and by microscopic examination using the H & E stain and Warthin-Starry silver stain. RESULTS: Rapid urease test was positive in 55% of all inoculated mice. Definite histologic changes and the evidence of H. pylori colonization were observed in the H. pylori infected group. Significant infiltration of inflammatory cells was observed 6 weeks after the last inoculation and the level of serum IgG against H. pylori was increased from 2 weeks after the last inoculation. CONCLUSIONS: The H. pylori isolated freshly from Korean adults could colonize the stomach of BALB/c mice and induce pathologic alterations that mimics human gastric diseases. This model would facilitate the investigations for the pathogenetic mechanisms of H. pylori infection. PMID- 10461427 TI - Assessment of body composition using dual energy x-ray absorptiometry in patients with liver cirrhosis: comparison with anthropometry. AB - OBJECTIVES: The aim of this study was to evaluate changes of body composition in cirrhotic patients. Dual energy x-ray absorptiometry (DEXA) and anthropometry were used, and the values obtained were compared. METHODS: Mid-arm fat and muscle areas were calculated by anthropometry in 66 cirrhotic patients and 94 healthy controls. In 37 of the cirrhotic patients and 39 of the controls, fat mass, lean soft tissue mass and bone mineral contents were measured with DEXA. RESULTS: The number of cirrhotic patients with measured values below the fifth percentile of normal controls was 21 (31.8%) by mid-arm fat area, six (9.1%) by mid-arm muscle area, 15 (40.5%) by fat mass and 0 (0%) by lean soft tissue mass. The fat mass in cirrhotic patients was less than in controls, whereas lean soft tissue mass and bone mineral content were not different. Fat depletion was severe in Child-class C patients and with severe ascites. Mid-arm fat area and fat mass showed close correlation (r = 0.85, p < 0.01), but mid-arm muscle area and lean soft tissue mass showed poor correlation (r = 0.32, p < 0.05). CONCLUSION: Cirrhotic patients showed lower fat component, with preserved lean soft tissue mass and bone mineral content. In clinical practice, the measurement of mid-arm fat area was useful for the assessment of fat mass. PMID- 10461428 TI - The imbalance between coagulation and fibrinolysis is related to the severity of the illness and the prognosis in sepsis. AB - OBJECTIVES: The coagulation and fibrinolytic system appears to be activated by the septic process independently, leading to the syndrome of disseminated intravascular coagulation (DIC). In this study, we investigated the changes within the hemostatic system related to the severity of the illness and the prognosis in patients with sepsis. METHODS: Plasma thrombin-antithrombin III (TAT) and plasmin-alpha 2-antiplasmin (PAP) complexes were measured using ELISA methods in 32 patients with sepsis and 20 controls and were analyzed according to the APACHE III scores and survival of the patients. RESULTS: Plasma TAT and PAP in patients with sepsis were significantly higher than controls. Nonsurvivors showed greater levels of TAT (21.7 +/- 22.3 ng/mL) and lower levels of PAP (628.4 +/- 378.1 ng/mL) than survivors (TAT: 11.1 +/- 11.2 ng/mL; PAP: 857.1 +/- 364.1 ng/mL). The imbalance between coagulation and fibrinolysis described as TAT/PAP ratio was closely related with APACHE III scores in patients with sepsis (r = 0.47) and the TAT/PAP ratio in nonsurvivors was significantly higher compared with survivors (34.4 +/- 21.4 vs. 14.4 +/- 13.8). CONCLUSION: In sepsis, both coagulation and the fibrinolysis system are activated and the imbalance between coagulation and fibrinolysis predisposes to the hypercoagulation state and is closely related to the severity of the disease and the prognosis. PMID- 10461429 TI - A case of Sweet's syndrome in patient with dermatomyositis. AB - Sweet's syndrome (SS) has been reported as an association with malignant neoplasms and autoimmune diseases, e.g., Behcet's disease, Sjogren's syndrome, and rheumatoid arthritis. But dermatomyositis (DM), one of the rare autoimmune diseases, was not reported as an associated disease of SS. We describe an interesting case of SS associated with DM. Diagnosis was made by skin biopsy, and subsequent clinical resolution occurred after institution of prednisolone. PMID- 10461430 TI - Epigastric appendiceal abscess with spontaneous drainage into the stomach. AB - The appendiceal abscess is a common complication of acute appendicitis and usually is located in the right lower quadrant of the abdomen. An epigastric appendiceal abscess has never been reported at an unusual location. We experienced an unusual case of a 49-year-old man with an epigastric appendiceal abscess. Initially, this abscess was suspected to be a pancreatic abscess. Abdominal CT scan and barium enema demonstrated a hyperrotated cecum with an appendiceal abscess in the left upper quadrant of the abdomen. An gastroscopy revealed a small fistula-like lesion with purulent coating at the bulging posterior gastric wall. The abscess resolved spontaneously. We believe that the abscess drained into the stomach through a small fistula between the stomach and abscess cavity. There was no recurrence for over 6 months. PMID- 10461431 TI - Imported tertian malaria resistant to primaquine. AB - In Plasmodium vivax and Plasmodium ovale malaria, some of the liver stage parasites remain dormant. The activation of these dormant forms (called hypnozoite) can give rise to relapse weeks, months or years after the initial infection. To prevent relapses, a course of primaquine may be given as terminal prophylaxis to patients. Different strains of Plasmodium vivax vary in their sensitivity to primaquine and, recently, cases of relapse of Plasmodium vivax after this standard primaquine therapy were reported from various countries. We reported a case of primaquine resistant malaria which initially was thought to be relapsed caused by loss of terminal prophylaxis. PMID- 10461433 TI - A case of sternal insufficiency fracture. AB - We report a case of insufficiency fracture of the sternum in a 70-year-old female patient with a review of the literature. She complained of sudden onset chest pain and aggravating dyspnea. She has been managed with corticosteroid due to chronic obstructive pulmonary disease for 15 years. Diagnosis of sternal insufficiency fracture presented with thoracic kyphosis was made on the basis of absence of trauma history, radiologic findings of lateral chest radiograph, bone scintigraphy and chest computed tomography. Thoracic kyphosis and osteoporosis secondary to menopause, corticosteroid therapy and limited mobility due to chronic obstructive pulmonary disease were considered as predisposing factors of the sternal insufficiency fracture in this patient. PMID- 10461432 TI - A case of xanthogranulomatous cholecystitis. AB - Xanthogranulomatous cholecystitis (XGC) is an uncommon, focal or diffuse destructive inflammatory disease of the gallbladder that is assumed to be a variant of conventional chronic cholecystitis. A 36-year-old male was admitted to Chonnam National University Hospital with a 10-day history of right upper quadrant pain with fever. 15 years ago, he was first diagnosed as having hemophilia A, and has been followed up in the department of Hematology. Computed tomogram (CT) revealed a well-marginated, uniform, marked wall thickening of the gallbladder with multiseptate enhancement. Magnetic resonance imaging (MRI) demonstrated diffuse wall thickening of the gallbladder by viewing high signal foci with signal void lesions. After factor VIII replacement, exploration was done. On operation, the gallbladder wall was thickened and the serosa were surrounded by dense fibrous adhesions which were often extensive and attached to the adjacent hepatic parenchyma. There was a small-sized abscess in the gallbladder wall near the cystic duct. Dissection between the gallbladder serosa and hepatic parenchyma was difficult. Cross sections through the wall revealed multiple yellow-colored, nodule-like lesions ranging from 0.5-2 cm. There were also multiple black pigmented gallstones ranging from 0.5-1 cm. The pathologic findings showed the collection of foamy histiocytes containing abundant lipid in the cytoplasm and admixed lymphoid cells. Histologically, it was confirmed as XGC. We report a case with XGC mimicking gallbladder cancer in a hemophilia patient. PMID- 10461434 TI - Swallow syncope. AB - Swallowing (or deglutition) syncope is an uncommon, vagally-mediated etiology for syncope that may be seen in children and adults. The mechanism of syncope involves afferent impulses from the upper gastrointestinal tract and efferent impulses to the heart that can produce a variety of bradyarrhythmias with atrioventricular block. Two cases of swallow syncope are reported, one associated with drinking a cold carbonated beverage, and the other precipitated by eating a large bolus of food (which we have termed "Vaso-Bagel" syncope). In evaluating patients with syncope, a history of a temporal relationship to eating or drinking should be sought. While changes in eating habits may be effective in some cases, permanent pacemaker placement is often indicated and is curative. PMID- 10461435 TI - Enhanced external counter pulsation: the Howard County experience in the first 18 patients. AB - Enhanced external counter pulsation is a noninvasive therapy that uses sequentially inflated pneumatic cuffs on the lower extremities to enhance coronary artery diastolic flow and decrease left ventricular afterload. We studied its effect on 18 patients with persistent angina despite maximal medical, surgical and catheter-based interventions. After enhanced external counter pulsation all subjects improved their functional class. Treatment with enhanced external counter pulsation improved functional class significantly from baseline 3.1 +/- 0.6 to 1.6 +/- 0.5 (p < .001). These results are consistent with the national experience. Growing physician awareness, recent Medicare approval status, and subsequent reimbursement will hopefully increase the use of this therapy. PMID- 10461436 TI - Carotid endarterectomy in a community hospital setting: a three-year experience. AB - The need for all medical institutions that treat carotid artery occlusive disease to continuously monitor their surgical experience is quite evident. Nonetheless, a national survey in 1995 found that only 15% of physicians reported knowing the perioperative stroke rate at the hospitals where they perform or refer patients for carotid endarterectomy. Neurologic morbidity related to the performance of carotid endarterectomy has been continuously monitored at Howard County General Hospital for the past decade, but the true significance of this data could not be appreciated without a more comprehensive analysis of the clinical experience. The present study was undertaken with the intent of providing this information. PMID- 10461437 TI - Laparoscopic spinal fusion. AB - We have reviewed the history of laparoscopic fusions and our results over the past two years, utilizing a femoral ring allograft and bone graft in 51 patients. The rate of fusion in this series is nearly 100%, with relief of radicular pain in about 90%. The average hospital stay is approximately two days and return to work is about six weeks. Minimally invasive techniques for spinal surgery will evolve with time and will become the standard of care in the near future. PMID- 10461438 TI - Ultrasonic assisted liposuction. AB - Ultrasonic assisted liposuction is a new and highly advanced technology that uses sound waves to emulsify body fat thereby allowing an efficient removal compared with the mechanical disruption of fat used in traditional liposuction. Safe application requires an exceptionally sophisticated plastic surgical, anesthetic, and operating room team. The media bombards patients with "magic wand" medical technology but seems less inclined to portray limitations, complications, and deaths. As with any emerging technology, ultrasonic assisted liposuction is producing its share of problems. This article provides an overview of this procedure and guidelines for patient selection. A conservative approach is emphasized for safety. PMID- 10461439 TI - Skin cancer detection and prevention: a community program promoting sun safe behaviors. AB - New cases of skin cancer will be diagnosed over one million times in the United States in 1999. Basal and squamous cell types can cause significant cosmetic morbidity. Malignant melanoma can cause serious morbidity and mortality. Thus, early detection and prevention efforts are crucial for all skin cancers. Studies have identified genetic, hereditary, and environmental factors associated with increased risk of developing skin cancer. In this article, the incidence and risk factors are reviewed. The Howard County Skin Cancer Awareness Project, established in 1996, is presented as a community based program that aims to increase public awareness about the prevention and early detection of skin cancer. Future directions are also addressed. PMID- 10461440 TI - Pentoxifylline treatment for microangiopathic hemolytic anemia caused by mechanical heart valves. AB - The use of pentoxifylline increased the hematocrit and eliminated the transfusion requirement for a patient who had microangiopathic hemolytic anemia due to three prosthetic heart valves. PMID- 10461441 TI - Bird fanciers' lung: a case report. AB - The clinical course of a case of bird fanciers' lung is described along with the considerations that are important in the management of this form of hypersensitivity pneumonitis. Diagnostic parameters, therapeutic considerations, and environmental issues are discussed in the context of the current literature. PMID- 10461442 TI - Cat scratch encephalopathy. AB - Cat scratch disease is usually a self-limited illness associated with tender lymph nodes, fever, malaise, and fatigue. Lymphadenopathy usually resolves spontaneously within three to four months. Cat scratch disease can be atypical as indicated by the presentation of our patient. PMID- 10461443 TI - Postural instability in Parkinson's disease. AB - Persons with Parkinson's disease are at great risk of suffering traumatic injuries from falls. Intervention with physical therapy and the use of assistive devices are helpful in preventing falls. Unfortunately, many patients are not referred for these interventions until they have already suffered traumatic injury. A simple measure of balance, which can easily be performed in an office setting, is the functional reach. This measure has been shown to be predictive of falls in the elderly. In this study, the functional reach was measured in patients with idiopathic Parkinson's disease during each visit over at least one year. Patients deemed at risk of falling were referred for physical therapy and possibly assistive devices. Those patients subsequently suffering falls were noncompliant with the recommendations. PMID- 10461444 TI - Physician volunteers. AB - There is an increasing population of working poor in our community. They earn too little to afford health insurance, yet they don't qualify for government assistance. Physician volunteers Howard County have joined together and developed a free clinic to meet this challenge. PMID- 10461445 TI - Infant mortality statistics from the 1997 period linked birth/infant death data set. AB - OBJECTIVES: This report presents 1997 period infant mortality statistics from the linked birth/infant death data set (linked file) by a wide variety of maternal and infant characteristics. METHODS: Descriptive tabulations of data are presented. RESULTS: In general, mortality rates were lowest for infants born to Asian and Pacific Islander mothers (5.0), followed by white (6.0), American Indian (8.7), and black (13.7) mothers. Infant mortality rates were higher for Puerto Rican mothers (7.9) than for Mexican (5.8), Cuban (5.5), Central and South American (5.5), or non-Hispanic white mothers (6.0). Infant mortality rates were higher for those infants whose mothers began prenatal care after the first trimester of pregnancy, were teenagers or 40 years of age or older, did not complete high school, were unmarried, or smoked during pregnancy. Infant mortality was also higher for male infants, multiple births, and infants born preterm or at low birthweight. In 1997, 65 percent of all infant deaths occurred to the 7.5 percent of infants bom at low birthweight. The three leading causes of infant death--Congenital anomalies, Disorders relating to short gestation and unspecified low birthweight (low birthweight), and Sudden infant death syndrome (SIDS) taken together accounted for nearly one-half of all infant deaths in the United States in 1997. Cause-specific mortality rates varied considerably by race and Hispanic origin. For black mothers, the infant mortality rate for low birthweight was four times that for white mothers. For American Indian mothers, the SIDS rate was 2.4 times that for white mothers. For Hispanic mothers, the SIDS rate was one-third lower than that for non-Hispanic white mothers. PMID- 10461446 TI - [Clinical aspects of heparin induced thrombocytopenia]. AB - Heparin induced thrombocytopenia has remained the major complication of therapy or prophylaxis with heparin. Although low molecular weight heparins seem to confer much lesser chance to induce thrombocytopenia, the danger is still considerable, and the fatal outcome is not rare. A lot of new data have been published about the origin, binding, physicochemical properties of the antibodies, the responsive platelet membrane receptors, and laboratory diagnosis in particular, however, many issues are still unresolved. The anticoagulant treatment of cases, in which heparin induced thrombocytopenia in associated with progressive, frequently arterial thrombosis still needs great skills, experience and the use of new generation antithrombotic agents. This review summarizes briefly the internationally accepted standard diagnostic and therapeutic protocols with heparin induced thrombocytopenia. PMID- 10461447 TI - [Prognostic value of the presence of the mutation of the codons 12, 13 and 61 in K-ras oncogene in colorectal cancer]. AB - Despite of extensive and intensive investigations, the predictive and prognostic value of c-K-ras mutation is not unequivocal. There has been reported about investigation the occurrence of mutations in the 88 colorectal cancer patient's specimen using polymerase chain reaction. Age: 61.9 years (27-80), gender 8 male, 42 female. Dukes' stages: 43 at the B, 35 at C, 10 at D. Primary of tumour: 52 colon, 36 rectal adenocarcinoma. Mutation out of one of the three ras-codons was detectable in the 54 cases, more frequently at the stage Dukes' C (p < 0.05). The ras-mutation concerned to more elevated death-rate in the stages Dukes' B and C (p < 0.01). Mean survival time to progression was significantly longer at the stage Dukes' B if mutation had not been detected (p < 0.01). The occurrence of the rate of genetic alteration was significantly more frequent at tumours of right-side colon, than left side (p < 0.02) or rectum (p < 0.05) one's. However, at the age of 41-50 years it was significantly more presented at the cases of rectal cancer (p < 0.01). At the age of 51-60 years mutations were detected among men at higher rate (p < 0.05). The cases of local recurrences concerned by mutation at the codon of 13 (p < 0.05). Occurrence of ras-oncogene is the sign of extremely malignant potential of tumour. This fact manifested itself in the time to progression and mean survival time of patients at same clinical or pathological stage. The higher frequency of genetic alterations at the proximal colon may be the reason of more unfavourable prognosis of the disease localised to this site. Reconstructing the molecular events, the presence of ras mutation can serve as a basis for prognosis of the disease and permit of potentially individualised therapeutic intervention. PMID- 10461448 TI - [Effectiveness of ceftibuten++ in the sequential therapy of respiratory and urinary tract infections]. AB - A prospective, comparative, multicentre trial was performed to study the efficacy and safety of ceftibuten in respiratory and complicated urinary tract infections. Patients (n = 152) requiring parenteral 2nd or 3rd generation cephalosporine therapy were randomly assigned to continue parenteral therapy (Group A) or to receive oral ceftibuten 400 mg, or 9 mg/kgbw/day (Group B) from the 3.-5. days on. The patients, whose conditions have not improved significantly at day 3-5, were omitted from the study, so the number of evaluated patients was 131. In Group A, out of 59 patients 51 were clinically cured, the bacteriological eradication rate was 47/54. In Group B, out of 72 patients 67 were cured and 62 out of 66 pathogens were eradicated. The cost of step-down therapy was 44.3% less than the parenteral one. No adverse effect was observed that could surely be attributed to ceftibuten. According to these data, ceftibuten can be used in step down therapy in respiratory and urinary tract infections requiring parenteral therapy at first and that offers a safe and less expensive therapeutic approach. PMID- 10461449 TI - [Cerebral uptake and metabolism of (11C) Vinpocetine in monkeys: PET studies]. AB - Vinpocetine, a vinca alkaloid, is a therapeutic agent widely used in the treatment of acute and chronic stroke patients. To explore the uptake and distribution of vinpocetine in the primate brain, vinpocetine was labelled with 11C and positron emission tomography (PET) was used to measure the uptake and distribution of 11C-vinpocetine in the brain and the trunk of a cynomolgous monkey. HPLC was used to determine the concentration of vinpocetine and its labelled metabolites in blood and plasma. Following the radioligand's intravenous administration, after an initial peak, the total concentration of radioactivity in blood was relatively stable with time. The uptake of 11C-vinpocetine into the brain was rapid and about 5% of the total injected radioactivity was present in the brain two minutes after drug administration. These facts indicate that the compound passes the blood-brain barrier readily and enters the brain. The radioactivity uptake was heterogeneously distributed among brain regions. The highest concentrations were found in the thalamus, the basal ganglia and certain neocortical regions. In an earlier PET investigation on chronic stroke patients the highest increases in cerebral blood flow and glucose metabolism after intravenous vinpocetine treatment occurred in these anatomical structures. The heterogenous regional distribution of vinpocetine and the observation that the highest uptake values in brain structures go parallel with the greatest regional blood flow and glucose metabolic rate increases indicate that direct CNS effects of vinpocetine should be considered as an explanation for the therapeutic effects. The confirmation of this suggestion requires further investigations. PMID- 10461450 TI - [Commemoration of Lajos Markusovszky--1999]. PMID- 10461451 TI - [From Vesalius to the electronic library]. PMID- 10461452 TI - [Communication from the Szent Istvan Municipal Hospital. Data on the surgical management of scarified pyloric stenosis (historical article)]. PMID- 10461453 TI - Lysosomes are sites of fluoroquinolone photosensitization in human skin fibroblasts: a microspectrofluorometric approach. AB - The fluoroquinolone antibiotics are widely used despite their strong phototoxicity under solar UV irradiation. Although they are known as good photodynamic photosensitizers, other factors than production of activated oxygen species may play a role in the effectiveness of the phototoxic effect. Subcellular localization is one of the important parameters that may determine this strength. Using microspectrofluorometry, it is shown that norfloxacin, ofloxacin, lomefloxacin, ciproflaxin and BAYy3118 are readily incorporated into lysosomes of HS68 human skin fibroblasts although weak staining of the whole cytoplasm also occurs especially with norfloxacin. Consistent with their photoinstability in solutions, the fluoroquinolones under study are readily photobleached by UVA in the HS68 fibroblasts. The BAYy3118 derivative that has the fastest bleaching rate also shows the strongest phototoxicity toward HS68 fibroblasts. Photosensitization with these fluoroquinolones induces lysosomal membrane damage as shown by the increased rate of leakage of the lysosomal probe lucifer yellow as compared to that observed with untreated cells. PMID- 10461454 TI - Photodynamic crosslinking of proteins. III. Kinetics of the FMN- and rose bengal sensitized photooxidation and intermolecular crosslinking of model tyrosine containing N-(2-hydroxypropyl)methacrylamide copolymers. AB - As part of a study on the role of Tyr residues in the photosensitized intermolecular crosslinking of proteins, we have surveyed the kinetics of the rose bengal- and flavin mononucleotide (FMN)-sensitized photooxidation and crosslinking of a water-soluble N-(2-hydroxypropyl)methacrylamide copolymer with attached 6-carbon side chains terminating in tyrosinamide groups (thus the -OH group of the Tyr is free, but both the amino and carboxyl groups are blocked, simulating the situation of a nonterminal Tyr in a protein). The intermolecular photodynamic crosslinking of the Tyr copolymer can result only from the formation of Tyr-Tyr (dityrosine) bonds, because the copolymer itself is not photooxidizable. Rose bengal, primarily a Type II (singlet oxygen) sensitizer, sensitized the rapid photooxidation of the Tyr residue in the Tyr copolymer only at high pH, where the Tyr phenolic group is ionized; crosslinking did not occur with rose bengal under any of the reaction conditions used. In contrast, FMN, which can sensitize by both Type I (free radical) and Type II processes, sensitized the photooxidation of the Tyr copolymer over the pH range 4-9.5. Also, significant photocrosslinking occurred, but only from pH 4 to 8, with a maximum rate at pH 6. Crosslinking required the presence of oxygen. Studies with inhibitors, D2O as solvent, catalase and superoxide dismutase indicated that the photooxidation and photocrosslinking of the Tyr copolymer with FMN at pH 6 were not mediated by singlet oxygen, superoxide or hydrogen peroxide. It appears that crosslinking involves the abstraction of an H atom from the Tyr phenolic group to give Tyr and FMN radicals. The Tyr radical in one Tyr copolymer can then react with a Tyr radical in another Tyr copolymer to give an intermolecular dityrosine crosslink. PMID- 10461455 TI - Femtosecond two-photon excited fluorescence of melanin. AB - Fluorescence of synthetic melanin in dimethyl sulfoxide has been excited by two photon absorption at 800 nm, using 120 fs pulses with photon flux densities > or = 10(27) cm-2 s-1. The shortest main component of the three-exponential decay of fluorescence is 200 +/- 2 ps. The overall spectral shape is red-shifted with respect to the 400 nm excited fluorescence. Two-photon excited melanin fluorescence also has been measured from excised samples of healthy human skin tissue. Because of the selectivity of melanin excitation via resonant two-photon absorption, it is hypothesized that fluorescence excited in this way may yield information on malignant transformation. PMID- 10461456 TI - Photophysical studies of A2-E, putative precursor of lipofuscin, in human retinal pigment epithelial cells. AB - With age, human retinal pigment epithelial cells accumulate lipofuscin that can absorb photons in the visible range leading to light-induced damage and impaired vision. A putative precursor of lipofuscin, 2-[2,6-dimethyl-8-(2,6,6-trimethyl-1 cyclohexen-1-yl)-1E,3E, 5E,7E- octatetraenyl]-1-(2-hydroxyethyl)-4-[4-methyl-6 (2,6,6-trimethyl-1 - cyclohexen-1-yl)-1E,3E,5E-hexatrienyl]-pyridinium (A2-E), has recently been isolated and characterized from aged human retinal pigment epithelial cells. We have found that A2-E inhibits the growth of human retinal pigment epithelial cells at concentrations greater than 1 microM. Time-resolved fluorescence measurements of 1 microM A2-E in solution, performed under 413 nm excitation, showed that fluorescence wave forms integrated across the spectrum (450-600 nm) were best-fitted with three decay times in the nanosecond and subnanosecond time scale: 6.6, 1.9 and 0.33 ns. Untreated retinal pigment epithelial cells were characterized by three fluorescence lifetimes: 6.3, 1.7 and 0.35 ns. In retinal pigment epithelial cells treated with 1 microM A2-E, the fluorescence decay was significantly faster, with the marked presence (approximately equal to 30%) of a fourth short lifetime (0.12 ns). These fluorescence decay times for A2-E bound to human retinal pigment epithelial cells are similar to those of lipofuscin granules isolated from aged human retinal pigment epithelial cells. This similarity supports the hypothesis that A2-E is a precursor of lipofuscin and suggests that A2-E may play a role in the overall light damage associated with age-related retinal diseases. PMID- 10461457 TI - Oxidation of guanine in cellular DNA by solar UV radiation: biological role. AB - The formation of cyclobutane pyrimidine dimers (CPD) and 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodGuo) was investigated in Chinese hamster ovary cells upon exposure to either UVC, UVB, UVA or simulated sunlight (SSL). Two cell lines were used, namely AT3-2 and UVL9, the latter being deficient in nucleotide excision repair and consequently UV sensitive. For all types of radiation, including UVA, CPD were found to be the predominant lesions quantitatively. At the biologically relevant doses used, UVC, UVB and SSL irradiation yielded 8-oxodGuo at a rather low level, whereas UVA radiation produced relatively higher amounts. The formation of CPD was 10(2) and 10(5) more effective upon UVC than UVB and UVA exposure. These yields of formation followed DNA absorption, even in the UVA range. The calculated relative spectral effectiveness in the production of the two lesions showed that efficient induction of 8-oxodGuo upon UVA irradiation was shifted toward longer wavelengths, in comparison with those for CPD formation, in agreement with a photosensitization mechanism. In addition, after exposure to SSL, about 19% and 20% of 8-oxodGuo were produced between 290-320 nm and 320-340 nm, respectively, whereas CPD were essentially (90%) induced in the UVB region. However, the ratio of CPD to 8-oxodGuo greatly differed from one source of light to the other: it was over 100 for UVB but only a few units for UVA source. The extent of 8-oxodGuo and CPD was also compared to the lethality for the different types of radiation. The involvement of 8-oxodGuo in cell killing by solar UV radiation was clearly ruled out. In addition, our previously reported mutation spectra demonstrated that the contribution of 8-oxodGuo in the overall solar UV mutagenic process is very minor. PMID- 10461458 TI - The soluble eumelanin precursor 5,6-dihydroxyindole-2-carboxylic acid enhances oxidative damage in human keratinocyte DNA after UVA irradiation. AB - Soluble melanin precursors are present in serum and may act as skin chromophores contributing to UVR-induced oxidative damage. Our study aimed to determine whether the soluble eumelanin precursor 5,6-dihydroxy-indole-2-carboxylic acid (DHICA) photosensitizes DNA damage in human keratinocytes exposed to UVA irradiation. The HaCaT keratinocytes were incubated with and without DHICA, before irradiation with broadband UVA (320-400 nm). The DNA photodamage was assessed using the comet assay that detects frank single-strand breaks (SSB) and specific oxidative lesions with the addition of endonuclease III. Without DHICA incubation, there was no significant increase in SSB, compared to unirradiated cells, for doses up to 48.5 J/cm2 (< 1 minimum erythemal dose). Preincubation with 0.5 microM DHICA caused an increase in SSB at every UVA dose (significant from 12.1 to 48.5 J/cm2), while varying the DHICA concentration (0.125-2 microM) showed this effect to be concentration dependent such that SSB increased and endonuclease III-sensitive sites decreased with increasing DHICA concentration. The irradiation of cells in the presence of antioxidants (catalase, mannitol and histidine) suggests that DHICA-induced photosensitization is mediated via singlet oxygen and, to a lesser extent, hydroxyl radicals. These results indicate that DHICA can induce strand breaks with UVA at clinically relevant doses via a mechanism involving reactive oxygen species. PMID- 10461459 TI - Early induction of binucleated cells by ultraviolet A (UVA) radiation: a possible role of microfilaments. AB - The effects of UVA (365 nm) radiation on the cellular distribution of F-actin and formation of binucleated cells have been studied using 3T3 Swiss albino mouse fibroblasts and V79 Chinese hamster fibroblasts. Ultraviolet A at biologically relevant fluences was found to disintegrate the actin filaments in the cells shortly (5 min) after irradiation, concomitant with the formation of cells with two nuclei. In 76-100% of the bi- and multinucleated cells the distribution of F actin was clearly altered. Cells in GI phase of the cell cycle were most probably involved in the formation of binucleated cells. The disintegration of F-actin was presumably not due to depolymerization of F-actin to G-actin, as the amount of F actin in the cells was unaltered after UVA exposure but rather due to direct breakage of the actin filaments. Ultraviolet B (297/302 nm) had no effect on the cellular distribution of microfilaments, not even at highly lethal fluences. PMID- 10461460 TI - Decreased host-cell reactivation of UV-irradiated adenovirus in human colon tumor cell lines that have normal post-UV survival. AB - An ongoing study in our laboratories is to examine the relationship of DNA repair defects to human cancer. Our underlying hypothesis has been that human tumors may arise that lack interesting DNA repair pathways if these pathways are important in preventing cancer. In this study, we found that the UV-irradiated adenoviruses showed hypersensitivity when assayed on monolayers of certain human colon tumor cell lines, including three that are reported to have defects in long patch DNA mismatch repair genes and one with no reported defect in mismatch repair. The survival curves showed two components. The first sensitive component was characteristic of 77-95% of the infections depending upon the cell line and the experiment and had an average slope indicating 4.8-fold hypersensitivity to UV. The average of the second-component slopes indicated that the remainder of the infections was accompanied by near-normal repair. Although the value of the first component indicated that the colon tumor lines supported the growth of UV-damaged adenoviruses poorly, the cell lines themselves showed the same post-UV colony forming ability as did normal human fibroblasts, and their ability to support the growth of N-methyl-N'-nitro-N-nitrosoguanidine-damaged adenoviruses was normal, i.e. it parallelled their ability to repair O6-methylguanine in vitro. We previously observed two-component survival curves when assaying UV-irradiated adenovirus on monolayers of all of seven strains of fibroblasts from Cockayne's syndrome patients. By contrast, single-component curves have been obtained using 21 strains of normal human fibroblasts and seven other tumor lines. We interpret the two-component survival curves in terms of the defective transcription-coupled repair of UV-induced DNA damage that is characteristic both of Cockayne's and certain colon tumor cell lines. In addition, four mismatch repair-deficient colon tumor lines were resistant to killing by elevated levels of dG. PMID- 10461461 TI - Effect of cyclobutane pyrimidine dimers on apoptosis induced by different wavelengths of UV. AB - Ultraviolet radiation within three different wavelength ranges, UVA (340-400 nm), UVB (290-320 nm) or UVC (200-290 nm), was shown to induce apoptosis in OCP13 cells, derived from the medaka fish. Morphological changes such as cell shrinkage and a decrease in the number of nucleoli appeared 4 h after UVA, UVB or UVC irradiation, although with different relative efficiencies. Doses required to induce apoptosis with similar efficiencies were about 2500-fold higher for UVA and 10-fold higher for UVB than for UVC. The following phenomena occurred after UVA irradiation but not after UVB or UVC irradiation. (1) Ultraviolet-A-induced cell detachment occurred with or without cycloheximide pretreatment. (2) Cells attached to plastic showed morphological changes such as rounding up of nuclei without a change in the cell distribution. (3) Morphological changes after UVA irradiation could not be evaded by photorepair treatment. (4) Morphological changes did not occur in cells attached to glass coverslips but only those in plastic dishes. (5) Apoptosis occurred without detectable increase of caspase-3 like activity. (6) Morphological changes were inhibited by N-acetylcysteine, a scavenger of active oxygen species. These results suggest the existence of two different pathways leading to apoptosis, one for long- (UVA) and the other for short- (UVB or UVC) wavelength radiation. PMID- 10461462 TI - Near-infrared fluorescence spectroscopy detects Alzheimer's disease in vitro. AB - The purpose of this study was to investigate whether near-infrared (NIR) fluorescence spectroscopy could be used to detect Alzheimer's disease (AD) by brain tissue autofluorescence. Unfixed temporal cortex specimens from AD cases and age-matched, non-AD controls were frozen at autopsy and then thawed just prior to spectral measurement. Spectra of intrinsic tissue fluorescence induced by 647 nm light were recorded from 650 to 850 nm. We used principal component analysis of the tissue spectra from 17 AD cases and 5 non-AD control cases in a calibration study to establish a diagnostic algorithm. Retrospectively applied to the calibration set, the algorithm correctly classified 23 of 24 specimens. In a prospective study of 19 specimens from 5 AD brains and 2 non-AD control brains, 3 of the 4 control specimens and all AD specimens were correctly diagnosed. Both the excitation light used and the measured brain tissue autofluorescence are at NIR wavelengths that can propagate through skull and overlying tissue. Therefore, our results demonstrate an optical spectroscopic technique that carries direct molecular level information about disease. This is the first step toward a clinical tool that has the potential to be applied to the noninvasive diagnosis of AD in living patients. PMID- 10461463 TI - Epidemiological support for an hypothesis for melanoma induction indicating a role for UVA radiation. AB - An hypothesis for melanoma induction is presented: UV radiation absorbed by melanin in melanocytes generates products that may activate the carcinogenic process. Products formed by UV absorption in the upper layers of the epidermis cannot diffuse down as far as to the melanocytes. Thus, melanin in the upper layer of the skin may be protective, while that in melanocytes may be photocarcinogenic. Observations that support this hypothesis include: (1) Africans with dark skin have a reduced risk of getting all types of skin cancer as compared with Caucasians, but the ratio of their incidence rates of cutaneous malignant melanoma to that of squamous cell carcinoma is larger than the corresponding ratio for Caucasians. (2) Albino Africans, as compared with normally pigmented Africans, seem to have a relatively small risk of getting cutaneous malignant melanomas compared to nonmelanomas. This is probably also true for albino and normally pigmented Caucasians. (3) Among sun-sensitive, poorly tanning persons, frequent UV exposures are associated with increased risk of melanoma, whereas among sun-resistant, well-tanning persons, increased frequency of exposure is associated with decreased melanoma risk. (4) It is likely that UVA, being absorbed by melanin, might have a melanoma-inducing effect. This is in agreement with some epidemiological investigations which indicate that sun-screen lotions may not protect sufficiently against melanoma induction. The relative latitude gradient for UVA is much smaller than that for UVB. The same is true for the relative latitude gradient of cutaneous malignant melanoma as compared with squamous cell carcinoma and basal cell carcinoma. Under the assumption that the average slopes of the curves relating incidence rates with fluences of carcinogenic UV radiation are similar for melanomas and nonmelanomas, these facts are in agreement with the assumption that UVA plays a significant role in the induction of melanomas in humans. This is in agreement with the experimental results with Xiphophorus. PMID- 10461464 TI - Chronic photodamage in skin of mast cell-deficient mice. AB - Solar elastosis is a hallmark of photoaged human skin and a prominent feature in experimentally produced photoaging in murine skin. The products of mast cells have been implicated in the development of photoaged skin. We evaluated whether products from mast cells mediate chronic UVB-induced changes in murine skin by employing a strain of mast cell-deficient mice, WWv. The responses in these mice were compared to those in BALB/c, another albino mouse strain. Mice were exposed three times per week to UVB radiation for 11 weeks; the total dose was 18.8 J/cm2. Irradiated WWv mice showed greater epidermal alterations than the irradiated BALB/c mice. In the dermis, a 3.6-fold increase in elastin content, as measured by desmosine, was produced in the UVB-treated BALB/c mice; in contrast, no difference was observed in elastin between UVB-treated and untreated WWv mice. Collagen content was not increased by UVB treatment in either strain, and the glycosaminoglycan content increased a similar amount in UVB-treated mice in both strains. The number of mast cells increased two-fold and the number of neutrophils increased six-fold in UVB-treated BALB/c mice compared to age-matched unirradiated controls. Neutrophils, as well as mast cells, were absent in untreated and UVB-treated WWv mouse skin. These results suggest that products of mast cells are important in the development of solar elastosis in murine skin either by directly inducing elastin production by fibroblasts or indirectly by mediating the presence of other cell types that produce products that increase fibroblast elastin production. PMID- 10461465 TI - Cloning and molecular characterization of the cDNA for the Brazilian larval click beetle Pyrearinus termitilluminans luciferase. AB - The larval click-beetle Pyrearinus termitilluminans elicits the phenomenon of luminous termite mounds in the central-west region of Brazil. The bioluminescence (BL) spectrum of this larva (lambda max = 534 nm) is one of the most blue-shifted reported among known luminescent Coleoptera. We have isolated mRNA from larval thoracic lanterns and constructed a cDNA library into a lambda ZAP II vector. An expression library was obtained after excision of the pBluescript plasmid. This library was screened by photodetection and one clone that emitted green BL (lambda max = 538 nm) was isolated. The 2.2 kb cDNA insert includes a 543 residue open reading frame showing 82% homology with the luciferase isoenzymes of Pyrophorus plagiophthalamus (Coleoptera: Elateridae). As expected, the region between residues 223 and 247 that contains the putative active site for BL color determination showed a higher degree of homology among click-beetle luciferases that elicit closer BL colors. The in vitro BL spectrum of recombinant P. termitilluminans luciferase also peaks at 538 nm and, as in the case of native enzyme, does not show any bathochromic shift upon decreasing the pH. PMID- 10461466 TI - Light-induced damage in the retina: differential effects of dimethylthiourea on photoreceptor survival, apoptosis and DNA oxidation. AB - In the rat, photoreceptor cell death from exposure to intense visible light can be prevented by prior treatment with antioxidants. In this study we subjected albino rats raised in dim cyclic light and rats made more susceptible to light damage by rearing in darkness to exposures of green light that led to similar losses of photoreceptor cells. Rhodopsin and photoreceptor DNA, indicators of the number of surviving photoreceptor cells, were determined at various times over a period of 14 days after light exposure. Fragmentation of DNA was determined over a similar time course by neutral and alkaline agarose gel electrophoresis. Apoptosis in retinal DNA was measured by quantitating the appearance of 180 base pair (bp) nucleosomal fragments. Oxidation of DNA was measured by electrochemical detection of the nucleoside 8-hydroxydeoxyguanosine (8-OHdG) after separation by high-performance chromatography. For albino rats reared in dim cyclic light, 24 h of intense light exposure resulted in the loss of 50% rhodopsin and photoreceptor cell DNA. In dark-reared rats, the losses were 40%, respectively, after only 3 h of intense light treatment. In both cases pretreatment with the antioxidant dimethylthiourea (DMTU) prevented rhodopsin and photoreceptor cell DNA loss. The kinetics of the light-induced apoptosis depended markedly on the rearing environment of the rats. The DNA ladders appeared within 12 h of the onset of intense light in the rats reared in dim cyclic light. In these rats the 180 bp fragment was at two-thirds of its maximum intensity immediately after 24 h of light exposure and reached the maximum 12 h later. Dimethylthiourea partially inhibited ladder formation in rats reared in dim cyclic light and delayed the time of appearance of the 180 bp maximum by 6 h. By contrast, in rats reared in darkness the 180 bp fragment was undetected immediately after 3 h of light exposure and reached its maximum 2 days later. Pretreatment with DMTU completely eliminated DNA ladders in these rats. Alkaline gel electrophoresis revealed a pattern of single-strand DNA breaks, with relatively high molecular weight fragments, 6 h after light exposure of dark-reared rats. Single-strand DNA breaks in cyclic light rats corresponded with the onset of apoptotic ladders, but peak values preceded by 12 h the peak of DNA ladder formation. The quantity of 8-OHdG in retinal DNA remained close to control values in all samples with the exception of a peak of twice the control value 18 h after light exposure in the dark-reared rats and a value 60% higher 16 days after exposure in cyclic light animals. Dimethylthiourea had no effect on the amount of oxidized purine in any of the samples. The differences between dark-reared rats and rats reared in dim cyclic light in the kinetics of DNA fragmentation and in their response to treatment with DMTU is consistent with previous observations of fundamental differences in retinal cell physiology in these animals. In dim light-reared rats, the pathway to apoptosis may be qualitatively different from the pathway to net photoreceptor loss in rats reared in darkness. The lack of effect of DMTU on 8-OHdG formation suggests that the oxidation of DNA bases is not a causal factor in light-mediated photoreceptor cell death. PMID- 10461467 TI - Sun exposure at school. AB - There is strong evidence that sun exposure during childhood and adolescence plays an important role in the etiology of skin cancer, in particular cutaneous melanoma. Between the age of 6 and 18, most children and adolescents will spend around 200 days per year at school and may receive a substantial fraction of their daily total solar ultraviolet radiation (UVR) exposure while at school. This study estimated the average daily erythemally effective dose of 70 grade 8 students from a high school in Townsville during 5 school days in July 1998. Through UV measurements of shade locations at the school and a combination of frequency counts and a questionnaire of grade 8 students, it was possible to determine the fraction of solar UVR reaching under the shade structures during lunch breaks and routine outdoor activities. Also, a routinely operating UV Biometer provided the annual variation of the daily dose that was used to calculate exposure levels for the 70 students. Our results suggest that up to 47% of the daily total dose fell within the time periods where students were outdoors during school hours. For students not seeking shade structures during the breaks (which usually was the case when involved in sport activities such as basketball or soccer), the average daily dose could have been as high as 14 SED (standard erythemal dose). Using results from the questionnaire of 70 grade 8 students, their average annual dose while at school was 414 SED or 2 SED per school day. However, the distribution of average daily erythemal effective dose per grade 8 student over the whole year showed that on 31% of all school days in 1998, this dose was exceeded. Because most previous attempts to change arguably poor sun protective behavior of young Australian children and adolescents at school showed little success, one way of decreasing the amount of harmful UVR reaching unprotected skin is the more careful design of shade structures at schools. PMID- 10461468 TI - Twins and the genetic architecture of osteoarthritis. PMID- 10461469 TI - Virus-associated vasculitides. PMID- 10461470 TI - Animal models of the antiphospholipid syndrome. AB - The antiphospholipid antibody syndrome (APS) is characterized by thrombocytopenia, recurrent thromboembolic phenomena and recurrent fetal loss, in association with anticardiolipin antibodies (aCL) and/or lupus anticoagulant (LA). Owing to the ethical and practical restrictions of experimentation on humans, we have to look to animal experimentation to broaden our knowledge of the pathogenesis and management of APS. Work has been carried out predominantly on strains of naive mice in which APS has been induced, passively and actively, using autoantibodies, autoantigens and other antigens. Studies of autoimmune prone mice and naive rabbits are present in the literature, to a lesser degree. We review the various animal models of the pathogenesis of APS, whether spontaneous or induced, which have been developed over the years. Although several of the models have provided insights into the relationship between antiphospholipid antibodies and fetal loss, very few give guidance to explain the link with thrombosis. Novel or experimental therapeutic regimens have to be tested on appropriate animal models before any kind of human clinical trials may proceed. The regimens devised thus far are also reviewed. PMID- 10461471 TI - Hyaluronic acid in the treatment of osteoarthritis of the knee. AB - OBJECTIVES: We examined the efficacy, safety and patient satisfaction of intra articular hyaluronic acid (HA) in patients with osteoarthritis of the knee. METHODS: One hundred patients with mild to moderate osteoarthritis of the knee entered a randomized blind-observer trial of 6 months HA vs placebo. Primary efficacy criteria were pain on walking, measured with a visual analogue scale, and the Lequesne Index. RESULTS: For pain on walking, a significant difference in favour of HA was found for completed patients at week 5, the end of the course of injections, and at month 6, the end of the study (P = 0.0087 and P = 0.0049, respectively). Further analysis using the Last Observation Carried Forward (LOCF) also showed a significant benefit favouring HA at month 6 (P = 0.0010). For the Lequesne Index, a significant difference in favour of HA was found at week 5 (P = 0.030) and at month 2 (P = 0.0431), but this was only of borderline significance at month 4 (P = 0.0528). Patients' global assessment of efficacy favoured HA at month 6 (P = 0.012). Improvement in other secondary criteria was generally superior in the HA group compared to placebo both at week 5 and month 6. Adverse events, mainly local injection site reactions, occurred in both groups with equal frequency. CONCLUSIONS: The study demonstrated that five weekly intra-articular injections of sodium hyaluronate (Hyalgan) were superior to placebo and well tolerated in patients with osteoarthritis of the knee with a symptomatic benefit which persisted for 6 months. PMID- 10461472 TI - Safety of a home exercise programme in patients with polymyositis and dermatomyositis: a pilot study. AB - OBJECTIVES: To investigate whether a home exercise programme could safely be performed by patients with stable, inactive polymyositis (PM) and dermatomyositis (DM), regarding disease activity, muscle function, health status and pain. METHODS: Ten patients with reduced muscle function completed the study. A home exercise programme including exercises for strength in the upper and lower limbs, neck and trunk, for mobility in the upper limbs and moderate stretching was developed. The patients exercised for 15 min and took a 15 min walk 5 days a week during a 12 week period. Assessments included clinical evaluation of disease activity, serum creatinine phosphokinase (CPK) levels, magnetic resonance imaging (MRI) of the quadriceps, repeated muscle biopsy of the vastus lateralis, a muscle function index (FI), a walking test and a health status instrument (the SF 36) performed at the start of the study and after 12 weeks. RESULTS: After 12 weeks of exercise, there were no signs of increased disease activity as assessed clinically, by CPK values, MRI or muscle biopsy findings. On an individual basis, all patients improved regarding muscle function according to the FI, in six cases the improvement reached statistical significance (P < 0.05). A significant improvement regarding muscle function in the upper and lower limbs, walking distance and general health status was achieved. CONCLUSIONS: Our results indicate that this home exercise programme can be safely employed in patients with stable, inactive PM and DM, with beneficial effects on muscle function. PMID- 10461473 TI - Enhanced production of interleukin-6 (IL-6), oncostatin M and soluble IL-6 receptor by cultured peripheral blood mononuclear cells from patients with systemic sclerosis. AB - OBJECTIVES: To determine whether the spontaneous production of interleukin 6 (IL 6), oncostatin M (OSM), soluble IL-6 receptor (sIL-6R) and soluble gp130 (sgp130) from peripheral blood mononuclear cells (PBMC) is increased in patients with systemic sclerosis (SSc). METHODS: The culture supernatants of PBMC from patients with SSc (n = 33) and healthy controls (n = 20) were examined by enzyme-linked immunosorbent assay. RESULTS: The production levels of IL-6, OSM and sIL-6R were significantly higher in patients with SSc than in controls. However, sgp130 levels in supernatants from patients with SSc were not significantly elevated when compared with those from controls. Soluble IL-6R levels correlated significantly with the severity of pulmonary fibrosis in patients with SSc. CONCLUSIONS: The enhanced production of IL-6, OSM and sIL-6R from PBMC may cooperatively contribute to the disease process in SSc. In particular, enhanced sIL-6R production from PBMC may be related to the development of pulmonary fibrosis via enhancement of IL-6 signal transduction in SSc, since sIL-6R can act as an agonist of IL-6. PMID- 10461474 TI - Serum YKL-40 concentrations in patients with rheumatoid arthritis: relation to disease activity. AB - OBJECTIVE: YKL-40, also called human cartilage glycoprotein-39, is secreted by chondrocytes, synovial cells, macrophages and neutrophils. Studies have shown that YKL-40 is an autoantigen in rheumatoid arthritis (RA). We evaluated whether serum YKL-40 was related to disease activity in patients with RA. METHODS: Serum YKL-40 was determined by radioimmunoassay in 156 patients with RA during a 1 yr longitudinal study. RESULTS: Serum YKL-40 was increased in 54% of the patients with clinically active disease. Patients with clinically active disease initially who became inactive after 12 months had a significant decrease in serum YKL-40 ( 30%, P < 0.002) and patients who changed from inactive to active disease had an increase in serum YKL-40. Patients who remained active had unchanged serum YKL-40 during the study. Serum YKL-40 decreased rapidly (-24% after 7 days, P < 0.01) during prednisolone therapy, and more slowly in patients treated with methotrexate only (-15% after 60 days, P < 0.01). Patients with early RA (disease duration < 3 yr, n = 50) and a persistently elevated serum YKL-40 were at risk of radiological disease progression as determined by Larsen score. CONCLUSION: Serum YKL-40 varies according to disease activity in RA, but provides in some respect information different from conventional markers. Our previous studies are consistent with a local release of YKL-40 in the arthritic joint followed by a secondary increase in serum YKL-40. YKL-40 may prove to be a new tool for the study of disease activity and pathophysiology of RA. PMID- 10461475 TI - Extrapyramidal type rigidity in rheumatoid arthritis. AB - OBJECTIVES: We had noted cogwheel rigidity in a number of patients with rheumatoid arthritis (RA). Based on this finding, we aimed to investigate formally the presence of rigidity and cogwheeling in RA patients. Our secondary aim was to survey the co-existence of RA and Parkinson's disease (PD). METHODS: A total of 87 consecutive patients with a diagnosis of RA, 78 patients with PD and 67 otherwise healthy patients attending a dedicated headache clinic participated in the study. RESULTS: Rigidity was observed in 24% of RA, 60% of PD and 2% of headache patients. The frequency among the RA patients was significantly higher compared to that of patients with headache (chi 2 = 15.2; P = 0.00009). The frequency of PD among the RA patients was 2/87 (2.3%), while the frequency of RA among the PD patients was 6/78 (7.7%). CONCLUSION: Rigidity can be observed in approximately a quarter of patients with RA. PMID- 10461476 TI - Depressed proliferative responses by peripheral blood mononuclear cells from early arthritis patients to mycobacterial heat shock protein 60. AB - OBJECTIVES: T-cell responses to mycobacterial heat shock protein 60 (M.hsp60) have been implicated in the pathogenesis of adjuvant arthritis, but whether they play a role in rheumatoid arthritis (RA) is undefined. We therefore examined T cell responses to M.hsp60 and to other recall antigens in a cohort of patients with early RA and in healthy controls. METHODS: In vitro peripheral blood mononuclear cells' (PBMC) proliferative responses to antigen were measured by [3H]thymidine incorporation, and results correlated with clinical and laboratory features of disease. RESULTS: Whereas responses to the recall antigens tetanus toxin and purified protein derivative (PPD) were equivalent in the two groups, responses to both M.hsp60 and the Escherichia coli hsp60 were lower in the RA patients. These results could not be explained by either the higher prevalence of HLA-DR4 in the RA group, or the disease severity of the patients. CONCLUSION: In the light of results from the adjuvant arthritis model which suggest that arthritis may be ameliorated by the actions of an hsp60-reactive T-cell population, the lack of response to M.hsp60 in RA could contribute to disease persistence. PMID- 10461477 TI - A vascular basis for repetitive strain injury. AB - OBJECTIVE: The blanket term 'repetitive strain injury' (RSI) covers a wide variety of work-related clinical syndromes, most of which are localized lesions. However, some patients complain of diffuse forearm pain, a clinically distinct form of RSI, the aetiology of which is unknown. METHODS: Using Doppler ultrasound, we measured the vascular responses to muscular work in the radial artery in 13 patients with bilateral diffuse forearm pain, seven with unilateral diffuse pain and 19 controls with localized arm pain. RESULTS: We found that in diffuse forearm pain the radial artery is relatively constricted compared to the controls and fails to vasodilate with exercise, which suggests that diffuse forearm pain may be due to physiological claudication of the working forearm muscle. CONCLUSION: A possible explanation is inhibition of local endothelial nitric oxide function, and this may be an unusual secondary, but self perpetuating, pain condition which can follow other more specific, but chronic, arm pain syndromes in susceptible individuals. PMID- 10461478 TI - Identification of thiopurine methyltransferase (TPMT) polymorphisms cannot predict myelosuppression in systemic lupus erythematosus patients taking azathioprine. AB - OBJECTIVE: To determine whether the presence of polymorphisms associated with reduced or absent activity of thiopurine methyltransferase (TPMT), an enzyme involved in azathioprine metabolism, can predict side-effects, particularly myelosuppression, in patients taking this drug. METHODS: The TPMT genotype was determined in 120 patients with systemic lupus erythematosus (SLE) together with 15 patients with inflammatory bowel disease (IBD) and correlated with the effects of clinical exposure to azathioprine. RESULTS: TPMT polymorphisms were detected in eight patients. Severe marrow toxicity occurred in the single homozygote identified. Azathioprine was generally well tolerated, but 11 drug-associated neutropenias were detected. In only one of the 11 cases was a TPMT polymorphism identified. CONCLUSION: Homozygous TPMT deficiency was associated with severe marrow suppression. In the majority of cases, however, TPMT genotyping prior to azathioprine therapy would not have predicted myelosuppressive events and may augment, but not replace, regular blood monitoring. PMID- 10461479 TI - Evaluation of a new Apo-1/Fas promoter polymorphism in rheumatoid arthritis and systemic lupus erythematosus patients. AB - OBJECTIVE: We looked for an association between the MvaI polymorphism, a recently reported polymorphism on the promoter of the Apo-1/Fas gene, and rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) patients. METHODS: Two cohorts of Caucasian RA patients (total number = 185) and one cohort of SLE patients (n = 103) were studied. The MvaI polymorphism was typed by polymerase chain reaction and followed by MvaI digestion and gel electrophoresis. RESULTS: A skewed distribution of MvaI genotypes was found in the first cohort of RA patients (n = 103) compared to the controls, as a result of increased MvaI*2 and decreased MvaI*1 homozygosity. This skewed distribution of genotypes was also observed in RA patients with either early onset of disease or with systemic involvement or progressive disease (assessed by the presence of erosions). The frequency of the MvaI*2 allele was significantly increased in female patients (P = 0.035), patients with extra-articular involvement (P = 0.04) and patients with early onset (P < 0.01), compared to the normals. To confirm these findings, the MvaI polymorphism was also examined in a second cohort of RA patients (n = 82). The results in this cohort did not replicate the associations shown in the first cohort of RA patients. Part of this inconsistency could be attributed to different populations and different parameters collected and analysed. In SLE patients, frequencies of MvaI alleles were not statistically different to the controls. However, MvaI*2 homozygosity was significantly higher in SLE patients with photosensitivity (P = 0.03) or oral ulcers (P = 0.01) than in SLE patients without these features. CONCLUSION: The role of the Apo-1/Fas gene promoter MvaI polymorphism in RA and SLE is unclear and further substantiation in larger patient samples is needed. PMID- 10461480 TI - Prediction of maximal back muscle strength from indices of body mass and fat-free body mass. AB - OBJECTIVES: The purpose of this study was to examine, in healthy volunteers, the relationship between back extensor muscle strength and body size. METHODS: The maximal voluntary isometric back extensor strength of 456 volunteers, aged 18-42 yr and with no history of low back pain, was measured in a standing, slightly flexed forward posture. This was then correlated with two indices of body size: body mass and fat-free body mass. RESULTS: Significant linear relationships were observed between back muscle strength and each of the two indices of body size. There was a gender difference in both the slope and the intercept of the regression equations describing the relationships. There was no independent influence of age within the range studied. CONCLUSION: It was possible to establish predictive equations for back extensor strength based on body size which could be used to quantify strength 'deficits', for instance in patients with low back pain, and to prescribe submaximal target forces for use in endurance training and testing. PMID- 10461481 TI - Shoulder joint impairment among Finns aged 30 years or over: prevalence, risk factors and co-morbidity. AB - OBJECTIVES: To describe the prevalence, risk factors and consequences of shoulder joint impairment in the population. METHODS: A representative sample (n = 7217) of the Finnish population aged > or = 30 yr participated in a health examination survey (the Mini-Finland Health Survey). The design of the survey allowed an independent assessment of disability, reported shoulder pain, shoulder joint impairment and major chronic co-morbidity. RESULTS: Shoulder impairment was observed in 8.8%, while pain was reported by 30%. The prevalence of shoulder pain decreased among the elderly, whereas impairments increased up to 20% of those aged 75-80 yr. In addition to age, sex, previous injury to the shoulder joint and a history of physically heavy work, diabetes was associated with shoulder impairment (OR 1.6, 95% CI 1.2-2.1). Shoulder impairment was associated with disability (adjusted OR 2.0, CI 1.6-2.5). CONCLUSIONS: Shoulder impairment is an important component of ill health among the elderly, and cannot be reduced to reported pain alone. PMID- 10461482 TI - Gait variables: appropriate objective outcome measures in rheumatoid arthritis. AB - OBJECTIVE: To assess the reliability and responsiveness of gait speed, cadence and stride length at two self-selected speeds (SSS) in subjects with rheumatoid arthritis (RA). METHODS: Thirty-one subjects with RA were assessed on three occasions. At each assessment session, subjects were asked to self-report walk pain on visual analogue scales, rate physical function using the Health Assessment Questionnaire, and walk five times at both a normal SSS and a fast SSS along an 8 m electric footswitch walkway. RESULTS: Despite stability of pain and physical function, there were significant gait changes from the first to the second assessment session at the normal SSS, although this 'learning' effect was not evident at either SSS between the second and the third assessment session. CONCLUSION: If the recommended protocol is followed, quantitative gait variables can provide reliable and responsive outcome measures in this population for use in evaluating therapeutic interventions. PMID- 10461483 TI - Extra-articular rheumatoid arthritis: prevalence and mortality. AB - OBJECTIVE: The prevalence and distribution of extra-articular manifestations of rheumatoid arthritis (ExRA) and associated mortality were studied retrospectively in a cohort of RA patients admitted to University Hospital, Malmo, Sweden, during the period 1990-94. RESULTS: Of 489 patients who fulfilled the 1987 ACR criteria for RA, 37 manifested onset of ExRA, predominantly serositis and cutaneous vasculitis, during the period, corresponding to a cumulative incidence of 7.9%. The occurrence of ExRA was independent of disease stage. Among patients with ExRA, 1 death/4.3 person-years at risk (pyr) occurred, as compared with 1 death/11.4 pyr in the non-ExRA subgroup. The age- and sex-adjusted mortality rate ratio was 2.49 (95% confidence interval 1.43-4.03). The major cause of death among ExRA cases was heart disease, which occurred in 9/13 cases (69%) in comparison to the expected 2.4 cases. CONCLUSION: In this series, serositis and cutaneous vasculitis were predominant extra-articular manifestations of RA; and mortality was greater in the ExRA than in the non-ExRA subgroup, perhaps due to a high frequency of associated heart disease. PMID- 10461484 TI - Anticardiolipin antibodies, free protein S levels and thrombosis: a survey in a selected population of rheumatoid arthritis patients. AB - OBJECTIVE: To investigate plasma levels of natural anticoagulant proteins such as protein S, protein C and antithrombin III in a selected population of patients with rheumatoid arthritis (RA) with and without anticardiolipin antibody (aCL) positivity, and to evaluate the possible relationships with an increased risk of thrombotic events in RA. METHODS: A total of 184 female RA patients attending our Extra-Articular Involvement RA Clinic were evaluated for aCL levels, total and free protein S, protein C and antithrombin III concentrations, and for the occurrence of thrombotic events. Patients were grouped as aCL positive (n = 35) and aCL negative (n = 149). RESULTS: Higher rates of venous and/or arterial thromboses were diagnosed in patients with RA compared to controls (P = 0.01). In particular, lower free protein S levels were found in aCL-positive patients with RA compared to both aCL-negative patients and controls (P = 0.001). Functional assays for protein C, antithrombin III as well as total protein S levels were found to be in the normal range in all patients and controls. CONCLUSION: The association observed between aCL positivity and decreased levels of free protein S in RA patients may represent one of the risk factors for thrombotic events. PMID- 10461485 TI - Sleep apnoea caused by rheumatoid arthritis. PMID- 10461486 TI - Re: Assessment of disease activity in rheumatoid arthritis using magnetic resonance imaging: quantification of pannus volume in the hands. PMID- 10461487 TI - Relevance of tumour necrosis factor alpha gene polymorphism in rheumatic disease. PMID- 10461488 TI - Wegener's granulomatosis and diabetes insipidus. PMID- 10461489 TI - Sjogren's syndrome: a community-based study of prevalence and impact--comment on the article by Thomas et al. PMID- 10461490 TI - Polytopic spondylodiscitis secondary to Streptococcus bovis endocarditis. PMID- 10461491 TI - Re: Estimating the prevalence of delayed median nerve conduction in the general population. PMID- 10461492 TI - Reality-based relative value scales. PMID- 10461493 TI - CNS Resident Award. Malignant meningiomas frequently lose multiple chromosomal regions in addition to 22q: putative meningioma progression loci identified by comparative genomic hybridization. PMID- 10461494 TI - Controversies in neurosurgery: microsurgery versus radiosurgery for arteriovenous malformations--introduction. PMID- 10461495 TI - Controversies in neurosurgery: microsurgery versus radiosurgery for arteriovenous malformations--the case for microsurgery. PMID- 10461496 TI - Radiosurgery versus surgery for arteriovenous malformations: the case for radiosurgery. PMID- 10461497 TI - Anterior cervical fusion: the case for fusion without plating. PMID- 10461498 TI - The argument for single-level anterior cervical discectomy and fusion with anterior plate fixation. PMID- 10461499 TI - Surgical or radiosurgical treatment for brain metastases? Opportunity lost, responsibility shirked. PMID- 10461500 TI - The case for radiosurgery. AB - Brain metastases represent a significant health-care problem, with almost 200,000 patient in the Unite States annually suffering from symptomatic parenchymal lesions. Lung, breast, melanoma, renal, and gastrointestinal cancers contribute the majority of lesions that come to clinical attention. Although median survival once brain metastases are diagnosed is less than a year, timely therapy can restore neurological function and can often prevent further neurological complications of cancer for the duration of a patient's survival. Important prognostic features associated with improved survival include the absence of extracranial disease progression, young age, a high pretreatment neurological status, one to three versus more than three lesions, and a long interval from primary disease diagnosis to the development of brain metastases. The need to aggressively treat brain metastases effectively is becoming increasingly important, however, as advances in the treatment of systemic disease result in an increasing number of patients developing brain metastases in the setting of limited systemic disease. For many such patients, surgery provides the best therapy, but results are still not encouraging because even patients with the best prognostic indicators often die within 18 to 24 months. Until Superior treatment modalities are developed, the judicious use of available techniques for treatment of patients with limited systemic disease provides the best opportunities for palliation and extended survival. Perhaps the most significant development in the treatment of patients with brain metastases during the last decade is the increasing use of radiosurgery. For patients with a single lesion, local control and survival rates of radiosurgery compare well with those produced with surgical resection. Radiosurgery remains an important treatment modality and, when used promptly, can reverse neurological deficits, often for the remainder of a patients life. There is compelling evidence to suggest that aggressive local therapy (surgery or radiosurgery) for patients with a single brain metastasis produces superior survival and quality of life compared with treatment with whole brain radiotherapy alone. However, surgery should be restricted to the minority of patients for whom brain metastases represents the life-threatening site of their disease. For an asymptomatic or mildly symptomatic patient with a lesion smaller than 3 cm in diameter, radiosurgery is an excellent alternative to surgery. Although radiosurgery is a noninvasive procedure, the same selection criteria should be considered as for those patients undergoing surgical resection. PMID- 10461501 TI - Surgical treatment of brain metastases. PMID- 10461502 TI - The management of pituitary adenomas: the MGH experience. PMID- 10461503 TI - Stroke: magnitude of the problem and impact on health care. PMID- 10461504 TI - Developing effective emergency systems for stroke. PMID- 10461505 TI - The neurosurgeon and the acute stroke patient in the emergency department: diagnosis and management. PMID- 10461506 TI - Protocols and critical pathways for stroke care. AB - We have presented a conceptual approach toward developing clinical protocols and critical pathways in a complex multidisciplinary environment with a commitment to clinical excellence, evidence-based practice methodology, and education. The process and the mood surrounding these have been more important in our view than any particular protocol or pathway. They have generated an attitude aiming toward avoidance of complications rather than crisis management. They have contributed to a philosophy of integrative multidisciplinary collaborations among various specialists, house staff, and nursing and paramedical personnel and a greater mutual sensitivity in interactions with medical center management and administration. The overall impact of this global approach has been quantifiable (Fig. 6), although the role(s) of one or more facets of it cannot be independently defined (21, 22). Protocols and clinical pathways should be viewed as components of total quality management. They should not be allowed to restrict the patient's or physician's choice of interventions, they should not inhibit in any way innovation or the introduction of novel methodologies. Yet, protocols and critical pathways should and do generate a pressure on every member of the health care team, a sense of negative entropy constantly urging a move toward a higher level of excellence and quality. PMID- 10461507 TI - Intensive care management of subarachnoid hemorrhage, ischemic stroke, and hemorrhagic stroke. PMID- 10461508 TI - Stroke: indications for emergent surgical intervention. AB - As the brain attack message is disseminated throughout our medical community and the awareness of the public increases, neurosurgeons will have the opportunity to treat patients with stroke at a much earlier time in the evolution of the process than we have been accustomed. Are the relatively unimpressive results of acute surgical intervention in patients operated on later in the course of the disease applicable to those who seek medical attention early, within the first few hours of ictus? There is little firm data. However, there is an overwhelming amount of anecdotal and experimental evidence supporting the potential for ultra-early intervention, which frequently should be surgical. New surgical techniques may improve safety and feasibility of emergent operations. In the coming years, diagnostic techniques such as perfusion/diffusion magnetic resonance imaging will allow the clinician to determine who may benefit from intervention. These determinations will be made on physiological data, addressing the issues of tissue viability and degree of compromise of the blood-brain barrier. In the future, the window of opportunity for intervention will not be solely a function of time from ictus or a qualitative impression based on collateral circulation as extrapolated from angiography, transcranial Doppler, or magnetic resonance angiography. These new magnetic resonance imaging techniques, which are beginning to be tested clinically or are still in the developmental stages, will provide the functional data now provided by positron emission tomography and xenon computed tomography, but with improved sensitivity, specificity, and logistical ease. Neurosurgeons have been leaders in stroke care and have provided some of the most important experimental rationale for the brain attack concept. These contributions include demonstration of the ischemic penumbra, the importance of time and potential collateral circulation as factors determining viability of ischemic tissue, and the value of early revascularization and many neuroprotective maneuvers in preserving brain tissue after arterial occlusion. There is every reason to preserve and to enhance the role of the neurosurgeon as a "stroke expert" and as a leading member of the brain attack team. Early access to patients with stroke will offer us the opportunity to test clinically, in a rigorous fashion, the value of surgical revascularization procedures (open or endovascular) and medical maneuvers that we have developed clinically and tested in the laboratory. We have shown, as we did with the bypass study, that neurosurgeons know how to perform these trials and abide by their results, even when they are not to our liking. PMID- 10461509 TI - Outcome science and stroke. PMID- 10461510 TI - Current patterns and future possibilities in the care of acute ischemic stroke. AB - Zack Hall, Director of the NINDS, succinctly described the present situation: "This is the golden age of neuroscience research" (verbal presentation at the annual meeting of the American Neurological Association, 1997). Recent trials in clinical research have demonstrated the power of thrombolytic therapy in acute ischemic stroke, and investigators across the country are now refining that therapy. The advantages of thrombolytic intervention can be provided to a much larger proportion of the population as we defined techniques to slow the rate of neuronal cell death after ischemia. However, the most exciting future opportunities for care of individuals of acute ischemic stroke arises from two somewhat unsuspected avenues. First, neuroscientists are learning in an extraordinarily rapid fashion the potentials of replacement of neural cell populations using progenitor cells. Second, the incredible explosion of genetic information has created an opportunity to identify genes responsible for atherosclerosis and other cerebrovascular disease. This, in turn, could lead to precise therapies that prevent or diminish the disease. Exploiting these opportunities requires that neural clinicians continue their cooperative efforts and, also, learn to work together with neuroscientists and geneticists. With such cooperation, we are poised to translate the golden age of neuroscience research into a genuine benefit for individuals with cerebrovascular disorders. PMID- 10461511 TI - Stroke: the neurosurgeon's domain. PMID- 10461512 TI - Neurosurgery of the spine in the year 2001: a four-year review. PMID- 10461513 TI - Gliomas--past, present, and future. PMID- 10461514 TI - Cerebral vasospasm: current clinical management and results. PMID- 10461515 TI - Historical perspective and future direction. PMID- 10461516 TI - Management of traumatic brain injury: past, present, and future. PMID- 10461517 TI - Present technology and future applications. PMID- 10461518 TI - Controversies in neurosurgery: summary and perspective. PMID- 10461519 TI - The case for surgery. PMID- 10461520 TI - The case for biopsy and adjuvant therapy. PMID- 10461521 TI - The case for resective surgery. PMID- 10461522 TI - Neurogenetic surgery: current limitations and the promise of gene- and virus based therapies. PMID- 10461523 TI - Extracranial carotid therapy: endarterectomy versus endovascular therapy. PMID- 10461524 TI - The case for stenting. PMID- 10461525 TI - The case for endarterectomy. PMID- 10461526 TI - Benign tumors of the cavernous sinus. PMID- 10461527 TI - The case for aggressive resection. PMID- 10461528 TI - Management of meningiomas of the cavernous sinus: conservative surgery and adjuvant therapy. PMID- 10461529 TI - Idiopathic interstitial pneumonitis/fibrosis: a historical note. PMID- 10461530 TI - Advances in radiology for interstitial lung disease. AB - Recently, with radiology some advances have been made in the care of patients with interstitial lung disease, predominately through the use of high-resolution computed tomography. This technique is helpful not only in the diagnosis of interstitial lung disease, but also in patient follow-up and management. Other techniques, including nuclear medicine and magnetic resonance imaging, have limited but well-defined uses. Positron emission tomography is a promising nuclear medicine technique, but its benefits have not been substantiated in the literature. PMID- 10461531 TI - The role of thoracic surgery in diagnosing interstitial lung disease. AB - In this article, the current status of thoracic surgery options for reaching a diagnosis in interstitial lung disease is described. When surgery is needed, mediastinoscopy is the first step in cases of suspected stage I or II sarcoidosis. If this is not the case, video-assisted thoracoscopy is currently preferred to open lung biopsy because the need for analgesia lessened, less blood is lost, the operative time is shorter, the complication rate is lower, and the postoperative stay is shorter. In some cases, video-assisted thoracoscopy may also be preferred to mediastinoscopy, especially in young women, for cosmetic reasons. PMID- 10461532 TI - Treatment issues in interstitial lung disease in tropical countries. AB - Management of interstitial lung disease, especially idiopathic pulmonary fibrosis, is both difficult and unsatisfactory. In many patients, only supportive therapy can be instituted. Attempts have been made to use anti-inflammatory therapy to reverse inflammation, provide symptomatic relief, stop disease progression, and prolong survival; the results of such treatment have varied from no improvement to significant prolongation of survival. Corticosteroids are the most frequently used anti-inflammatory agents. Cytotoxic drugs, such as oral azathioprine or intermittent intravenous cyclophosphamide, have also been shown to be effective both alone and in combination with low-dose oral corticosteroids. Of the other antifibrotic drugs that have been used, colchicine seems to provide some benefit. It is especially useful in aged persons and those with corticosteroid-induced problems or concomitant illnesses that are likely to be worsened by steroids. Anti-inflammatory therapy is costly to administer and monitor, particularly in the developing world. It is therefore important to consider these issues before instituting treatment. Younger patients and patients with less-severe disease of recent onset are most likely to respond to treatment. Similarly, patients with lymphocytic alveolitis or desquamative interstitial pneumonia respond better. Despite the use of newer strategies for treatment, the overall prognosis for patients with interstitial lung disease has not really changed, and the median population survival remains almost the same as it was about 30 years ago. PMID- 10461533 TI - Clinical courses and prognoses of pulmonary sarcoidosis. AB - In this article, long-term prognoses and prognostic factors of patients with sarcoidosis are reviewed. In patients with intrathoracic sarcoidosis, functional impairments and parenchymal lesions at the time of initial examination strongly predicted an unfavorable prognosis. We also discussed the significance of extrathoracic lesions in terms of clinical outcomes of intrathoracic sarcoidosis. In addition, we focused on the genetic approach and the new insights being offering with respect to the disease susceptibility and the development of pulmonary lesions. PMID- 10461534 TI - A new look at hypersensitivity pneumonitis. AB - Hypersensitivity pneumonitis is an immunologically induced lung disease. Although both immune complex-mediated immune response and T cell-mediated immune response are involved in the pathogenesis of the disease, recent studies show the latter mechanism to be more important. As for T cell-mediated immune response, Th1/Th2 and Tc1/Tc2 cytokines produced by CD4+ and CD8+ T cells play important roles in the development of granulomatous inflammation in the lung, a pathologically characteristic feature of the disease. The critical distinction between CD4+ and CD8+ T cells pertains to recognition of antigens presented by different major histocompatibility complex molecules. Serum levels of KL-6 and soluble intercellular adhesion molecule-1 in patients with HP are useful markers of the disease activity. The chronic form of HP can be difficult to diagnose, and provocation testing is helpful. Erythromycin might be useful for anti inflammatory therapy. PMID- 10461535 TI - Pulmonary involvement in adult-onset Still's disease. AB - Adult-onset Still's disease (AOSD) is a rare splenic disorder with an unknown cause. It is not uncommon for AOSD to involve other organs, such as the liver; the kidney; the bone marrow; and, less often, the lungs. In this review, we discuss the pulmonary complications of AOSD. Pulmonary involvement in AOSD usually consists of pleural effusion or transient pulmonary infiltrates, but it may become life threatening if it progresses to the adult respiratory distress syndrome. Chronic conditions, such as restrictive lung disease, have also been reported in patients with AOSD. The only treatment currently available is high dose steroids, although other agents, such as intravenous immunoglobulin, cyclophosphamide, and azathioprine, have been tried with some success. PMID- 10461536 TI - Pulmonary veno-occlusive disease. AB - Pulmonary veno-occlusive disease (PVOD) is a rare cause of pulmonary hypertension that mainly affects children and young adults. Its cause is unknown, although viral infections and drugs have been implicated. Patients with PVOD present with symptoms of right-sided heart failure. Radiologic examination shows prominent pulmonary arteries with Kerley B lines, pleural effusion, and mediastinal adenopathy. The definite diagnosis is made by histologic examination. Eccentric intimal fibrosis and recanalized thrombi in pulmonary veins and venules, arterialized veins, alveolar edema, and medial hypertrophy of arteries are seen on lung biopsy. No effective treatment is available; lung transplantation has been tried. The prognosis associated with PVOD is poor. PMID- 10461537 TI - Pulmonary involvement in Behcet disease. AB - Pulmonary artery aneurysms varying in size and number continue to be the principal feature of pulmonary involvement in Behcet disease (BD). Pulmonary aneurysms have been reported to be associated with cardiac thromboses, mainly in the right heart. Bronchiolitis obliterans organizing pneumonia has also been seen in the setting of pulmonary artery aneurysms. Aneurysms of the aortic arch and subclavian artery are rarely recognized thoracic manifestations of BD. Noninvasive imaging techniques, such as helical computed tomography and magnetic resonance angiography, are the safer and preferred methods for identifying aneurysms and thrombi. Digital substraction angiography has been found to be inadequate in showing thrombosed vessels. Follow-up computed tomography was performed in the documentation of the aneurysmal healing process. In patients receiving immunosuppressive treatment, intramural thrombus formation occurred and was followed by aneurysmal regression and disappearance. General principles for the treatment of systemic vasculitis are used in the treatment of BD. PMID- 10461538 TI - Paracoccidioidomycosis. AB - Paracoccidioidomycosis is a systemic infection caused by the dimorphic fungus Paracoccidioides brasiliensis. It is commonly an endemic disease in Latin America, but several cases have been reported outside this area, particularly now in this time of world globalization. Primary pulmonary infection occurs commonly in the first and second decades of life and usually has a benign, self-limited respiratory infection course. The adult chronic manifestation of the disease is usually the result of reactivation of quiescent lesions with diffuse lung infiltrates, predominately of the interstitial pattern, with or without involvement of various other organs. The finding of this disease in a patient is an important step for the large differential diagnosis of the interstitial lung diseases group. PMID- 10461539 TI - Pulmonary associations in familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) is a hereditary periodic fever syndrome expressed by acute episodes of fever and painful manifestations. In this report, the pulmonary manifestations of FMF are reviewed, the most prominent of which are chest attacks due to pleuritis. Nephropathic amyloidosis of the AA type, which complicates FMF in most untreated patients, may progress to affect other organs, including the lungs, but this rarely produces noticeable symptoms. The common association between FMF and vasculitis makes pulmonary hemorrhage, infarction, or infiltrates highly possible. These complications, however, have been reported only rarely. Asthma was found to occur less often than expected in patients with FMF, but methodologic faults make this finding doubtful. Finally, the occurrence of mesothelioma in five patients with FMF who were not exposed to asbestos suggests a role for recurrent FMF serositis in the pathogenesis of this malignancy. PMID- 10461540 TI - Bibliography. Current world literature. Interstitial lung disease. PMID- 10461541 TI - Food toxins, ampa receptors, and motor neuron diseases. AB - Environmental chemicals involved in the etiology of human neurodegenerative disorders are challenging to identify. Described here is research designed to determine the etiology and molecular pathogenesis of nerve cell degeneration in two little known corticomotoneuronal diseases with established environmental triggers. Both conditions are toxic-nutritional disorders dominated by persistent spastic weakness of the legs and degeneration of corresponding corticospinal pathways. Lathyrism, a disease caused by dietary dependence on grass pea (Lathyrus sativus), is mediated by a stereospecific plant amino acid (beta-N oxalylamino-L-alanine) that serves as a potent agonist at the (RS)-alpha-amino-3 hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) subclass of neuronal glutamate receptors. A neurologically similar disorder, konzo ("tied legs"), is found among protein-poor African communities that rely for food on cyanogen-containing cassava roots. Thiocyanate, the principal metabolite of cyanide, is an attractive etiologic candidate for konzo because it selectively promotes the action of glutamate at AMPA receptors. Studies are urgently needed to assess the health effects of cassava and other cyanogenic plants, components of which are widely used as food. PMID- 10461543 TI - Neurotoxic mechanisms of degeneration in motor neuron diseases. PMID- 10461542 TI - Damage and repair of nerve cell DNA in toxic stress. AB - It is generally agreed that ALS/PDC is triggered by a disappearing environmental factor peculiar to the lifestyle of people of the western Pacific (i.e., Guam, Irian Jaya, Indonesia, and the Kii Peninsula of Japan). A strong candidate is the cycad plant genotoxin cycasin, the beta-D-glucoside of methylazoxymethanol (MAM). We propose that prenatal or postnatal exposure to low levels of cycasin/MAM may damage neuronal DNA, compromise DNA repair, perturb neuronal gene expression, and irreversibly alter cell function to precipitate a slowly evolving disease ("slow toxin" hypothesis). In support of our hypothesis, we have demonstrated the following: 1. DNA from postmitotic rodent central nervous system neurons is particularly sensitive to damage by MAM. 2. MAM reduces DNA repair in human and rodent neurons, whereas DNA-repair inhibitors potentiate MAM-induced DNA damage and toxicity in mature rodent nervous tissue. 3. Human neurons (SY5Y neuroblastoma) that are deficient in DNA repair are susceptible to MAM-induced cytotoxicity and DNA damage, whereas overexpression of DNA repair in similar cells is protective. 4. MAM alters gene expression in SY5Y human neuroblastoma cells and, in the presence of DNA damage and reduced DNA repair, enhances glutamate-modulated expression of tau mRNA in rat primary neurons; the corresponding protein (TAU) is elevated in ALS/PDC and Alzheimer's disease. These findings support a direct relationship between MAM-induced DNA damage and neurotoxicity and suggest the genotoxin may operate in a similar manner in vivo. More broadly, a combination of genotoxin-induced DNA damage (via exogenous and/or endogenous agents) and disturbed DNA repair may be important contributing factors in the slow and progressive degeneration of neurons that is characteristic of sporadic neurodegenerative disease. Preliminary studies demonstrate that DNA repair is reduced in the brain of subjects with western Pacific ALS/PDC, ALS, and Alzheimer's disease, which would increase the susceptibility of brain tissue to DNA damage by endogenous/exogenous genotoxins. Interindividual differences in the extent of prior exposure to DNA-damaging agents and/or the efficiency of its repair might produce population variety in the rate of damage accumulation and explain the susceptibility of certain individuals to sporadic neurodegenerative disease. Studies are underway using DNA-repair proficient and deficient neuronal cell cultures and mutant mice to explore gene-environment interplay with respect to MAM treatment, DNA damage, and DNA repair, and the age-related appearance of neurobehavioral and neuropathological compromise. PMID- 10461544 TI - Toxic neuronal apoptosis and modifications of tau and APP gene and protein expressions. AB - The causes and the mechanisms of neuronal death in Alzheimer's disease are not elucidated, although some new insights have been proposed over the past years, including free-radical toxicity, beta-amyloid toxicity, excitotoxicity, and disturbed cellular calcium metabolism. Some authors have also pointed out that apoptosis could play a role in neuronal degeneration, but it is still largely debated. Here, we review some recent data linking the induction of experimental neuronal apoptosis in vitro and the molecular pathology of the tau protein and amyloid precursor protein (APP). In cultures exposed to mild glutamate toxicity, tau mRNA expression, not beta-actin, is enhanced in stressed neurons. The Guam cycad toxin metabolite methylazoxymethanol also produces an increase of tau gene transcription that exacerbates changes induced by glutamate. In serum-deprived cultures or glutamate-exposed cultures, neurons committed to apoptosis have a reduced tau gene expression, whereas resistant neurons display a stable or even augmented tau mRNA expression accompanied by a persistent tau phosphorylation near serine 202. In the same conditions, stressed neurons produce membrane blebbings strongly immunopositive for APP and putative amyloidogenic fragments that are subsequently released in the extracellular space. Experimental apoptosis in neurons can recapitulate tau and APP modifications that could be associated with a selective vulnerability and a progression of cellular degeneration along the neuronal network. PMID- 10461545 TI - FK506 and the role of the immunophilin FKBP-52 in nerve regeneration. AB - In summary, FKBP-12 does not mediate the neurite outgrowth-promoting properties of neuroimmunophilin ligands (e.g., FK506). Instead, the neurotrophic properties of neuroimmunophilin ligands (FK506) and steroid hormones are mediated by disruption of steroid-receptor complexes. It remains unclear which component mediates neurite outgrowth, although the most likely candidates are FKBP-52, hsp 90, and p23 [42]. Regardless of the underlying mechanism involved, the FKBP-52 antibody data reveal that it should be possible to design, based on the structure of FK506, non-FKBP-12-binding (nonimmunosuppressant) compounds selective for FKBP 52 and test these new libraries for their ability to augment nerve regeneration. It may also be possible to exploit the structure of geldanamycin to develop a new class of hsp-90-binding compounds for use in nerve regeneration. PMID- 10461546 TI - Metabolism-based drug interactions involving oral azole antifungals in humans. PMID- 10461547 TI - The role of CYP2B6 in human xenobiotic metabolism. PMID- 10461548 TI - Frontier orbitals in chemical and biological activity: quantitative relationships and mechanistic implications. PMID- 10461549 TI - Thrombolytic therapy has been advocated as an effective treatment for acute ischemic stroke. PMID- 10461550 TI - Analysis of time management in stroke patients in three French emergency departments: from stroke onset to computed tomography scan. AB - The aim of this study was to determine and identify the factors associated with shortening or lengthening time interval from stroke onset to performance of computed tomography (CT) scan in stroke patients admitted to three French emergency departments. All suspected stroke patients were eligible (n = 317). The time intervals between stroke onset and presentation to the emergency department and between CT scan request and CT scan performance were determined. Twelve variables likely to influence time interval before presentation to the emergency department, and five variables likely to influence time interval before CT scan performance were evaluated using stepwise regression analysis. Of the 317 patients included in the study, the mean time interval from stroke onset to CT scan performance for 180 patients was 7 hours 46 minutes (466 minutes). The mean time interval between stroke onset and presentation to the emergency department was 4 hours 36 minutes (276 minutes), varying according to the study site, level of initial severity, medical contact before admission, witnesses at stroke onset, and mode of transportation. The mean time interval between request and CT scan performance was 2 hours 14 minutes (134 minutes), varying upon the site, hour of CT scan request, type of stroke and level of severity at admission. It is concluded that current delays in stroke management are often incompatible with early treatment. The public needs to be informed and admission procedures reorganized. Improved response to the urgency of ischaemic stroke is required as well as direct access to the scanner during periods of scheduled use. PMID- 10461551 TI - Decreased health care quality associated with emergency department overcrowding. AB - The objective of this study was to assess the influence of overcrowding on health care quality provided by emergency departments (ED). The study was carried out in an urban, university tertiary care hospital. All patients seen at the internal medicine unit (IMU) of the ED who returned during the following 72 hours, and those who died in the ED rooms were included in the study. During a consecutive period of 2 years (104 weeks), we prospectively quantified the number of weekly visits, revisits and deaths. We calculated revisit and mortality rates (in respect of percentage of all visited patients) for each week. Correlation between the number of weekly visits, and revisit and mortality rates was assessed using a simple linear regression model. We consigned 81,301 visits, 1137 revisits and 648 deaths; mean (+/- SD) number of weekly visits, revisits and deaths were 782 (68), 10.93 (3.97) and 6.23 (3.04) respectively; weekly revisit rate was 1.40% (0.48%) and weekly mortality rate was 0.79% (0.36%). We observed a significant, positive correlation between mortality rates and weekly number of visits (p = 0.01). Although a similar trend was also found for revisit rates, such an increase did not reach statistical significance (p = 0.06). It is concluded that since revisit and mortality rates constitute good health care quality markers, present data demonstrate that ED overcrowding implies a decrease in the health care quality provided by it. PMID- 10461552 TI - Patients' attitudes to medical students in the accident and emergency department. AB - The objective of this study was to assess patients' attitudes towards medical students in the accident and emergency department. Patients attending a university department of accident and emergency were given a questionnaire asking about their attitudes to medical students. Ninety-three per cent of the patients would not object to being seen and examined by a medical student in the accident and emergency department. No patients reported feeling pressurized to accept this proposal. It is concluded that most patients attending the emergency department had favourable attitudes towards medical students. PMID- 10461553 TI - Advanced trauma life support (ATLS) courses: should training be refocused towards rural physicians? AB - Recently there has been an increase in the number of courses designed to improve the theoretical knowledge and practical skills of health workers for immediate emergency management under life-threatening conditions. However, the numbers of applicants for these courses far exceed the available places. Priorities should be reviewed to solve the current shortfall. The purpose of this study was to compare the improvement in basic knowledge associated with the advanced trauma life support (ATLS) programme for paediatricians practising in a metropolitan area, with those practising in rural hospitals. This should facilitate proper use of the limited existing resources for training of physicians. A retrospective, comparative analysis of the impact of the ATLS courses for pediatricians was assessed by analysing scores of multiple choice tests before and after the programme. The study group comprised all Israeli paediatricians (n = 72) who completed the course during 1996. Performance was compared between paediatricians working in major, level I (n = 39) and rural, level II trauma centres (n = 33). After the course, all paediatricians improved their scores; those in level I trauma centres improved their mean precourse scores from 72 +/- 13.6 to 85 +/- 5.4, while those working in level II trauma centres improved from 67 +/- 12.4 to 85 +/- 5.0. The impact of the course was reflected in the net and proportional gains. Both were higher for paediatricians working in level II trauma centres, compared with those working in metropolitan trauma centres. Furthermore, the net gain was significantly higher (p < 0.05). Efforts should be made to increase the participation of paediatricians practising in rural level II trauma centres in trauma care courses. Further research should address the applicability of these results for recruiting primary care physicians to the ATLS courses. PMID- 10461554 TI - The influence of the mode of emergency ambulance transportation on the emergency patient's outcome. AB - Emergency transport by an ambulance can cause considerable psychical and physical stress for patients. We determined the haemodynamic and endocrinological values of 54 healthy volunteers subjected to one high speed emergency transport and one smooth transport. There were significant differences in all measurements: heart rate (p < or = 0.001), blood pressure, cortisol (p < or = 0.01), prolactin, somatotropine and ACTH between the two modes of transportation. We hypothesize that the additional stress of high speed ambulance transport particularly in patients with acute cardiac disease may result in additional morbidity. PMID- 10461555 TI - Motor cortex excitability changes within 8 hours after ischaemic stroke may predict the functional outcome. AB - Motor evoked potentials after magnetic transcranial stimulation and the excitability of the motor cortex to increasing magnetic stimulus intensities were evaluated in six patients with hemiparesis after ischaemic stroke within 8 hours after stroke. The latencies of motor evoked potentials were normal in all patients. After stimulation of the ischaemic hemisphere we obtained responses comparable with the contralateral ones in two patients (mean NIH score 2 (SD 0)) and this group was completely asymptomatic after 15 days (NIH score 0). In four patients the excitability of the motor cortex involved by the ischaemia was reduced and magnetic motor threshold was higher than that of the spared motor cortex. This finding was associated with a poor motor recovery and the NIH score after 15 days was unchanged (NIH score 1.75 (SD 1.5)). The present data suggest that the evaluation of the excitability of motor cortex may offer a mean of predicting functional outcome following stroke. PMID- 10461556 TI - Thermodynamic modelling of hypothermia. AB - Hypothermia is an important threat in trauma patients. The prevention of accidental hypothermia requires a thermal steady state. A simple mathematical model describing thermal steady state is introduced. When applied to trauma patients the model seems sensitive to changes in temperature of intravenous fluids. To simulate possible strategies to prevent hypothermia in the trauma patient the mathematical solution needs to be extended to describe situations where steady state does not exist. From these simulations it can be concluded that infusion heating devices are mandatory in patients with high fluid requirements. PMID- 10461557 TI - Intravenous urography in the emergency department: when do we need it? AB - Intravenous urography (IVU) is a useful radiographic study in the detection of renal and ureteral calculi. However, it is time consuming, expensive, and exposes the patient to i.v. contrast and radiation. To determine the impact of utilizing IVU less for the detection of renal calculi, criteria for ordering IVU in the emergency department (ED) were evaluated, and patients with high probability of positive IVU were identified. Variables included presence of acute flank pain with haematuria, prior history of renal calculus, degree of haematuria, and uncontrolled pain. We reviewed patients presenting with acute flank and abdominal pain with haematuria from May 1995 to May 1996 at a large urban university hospital. Charts were abstracted for prior history, reason for ordering IVU, time in the ED, laboratory results, IVU result, final diagnosis, and disposition. Data was analysed with Student's t-test, Wilcoxon rank-sum and receiver operating characteristic (ROC) analysis. A total of 302 patients were identified, and 185 underwent IVU during the study period. For patients with prior history of renal calculi 82% had positive IVU (sensitivity 51%, specificity 87%). For patients with both acute flank pain and haematuria, 92% had a positive IVU (sensitivity 93%, specificity 43%), and 19% of patients with abdominal pain of unclear aetiology with haematuria had a positive IVU. All patients with uncontrolled pain had evidence of high-grade obstruction on IVU. Degree of haematuria was not predictive of a positive IVU from ROC curve derivation. IVU is a useful study in the ED but may be overutilized, leading to lengthy patient stays. The combined objective findings of acute flank pain and haematuria are sensitive, and prior history is specific in identifying patients with renal calculi. Degree of haematuria was not useful in predicting renal calculi. By utilizing the criteria of acute flank pain and haematuria as a decision aid, 66% of all IVUs ordered could have been avoided. PMID- 10461558 TI - Ruptured spleen due to blunt trauma in children: analysis of blood transfusion requirements. AB - Selective non-operative management of splenic injury in children is generally considered to be safe, and the majority of those with isolated injuries do not require blood transfusion. Eighty-four children were treated for blunt splenic trauma from 1988 to 1997 in the Department of Paediatric Surgery, The Medical Faculty of Uludag University, Bursa, Turkey. Management involved non-operative care in 56 cases (66.7%), splenectomy in 20 (23.8%) and splenorraphy in eight (9.5%) cases. There were no later complications related to splenic injury. The overall mortality was 9.5% (8/84) and the factors effecting mortality were additional intra-abdominal and/or other system injuries. Twenty-two children were not transfused during non-operative treatment. In our study, only 57.1% of the children in the non-operative group received blood transfusions. Some of the patients in this group received only small amounts of blood and probably would have recovered without it. It is concluded that, based on a very strict protocol in conservative management, the total amount of transfused blood could be reduced in children with splenic injuries due to blunt abdominal trauma. PMID- 10461559 TI - Order in chaos: modelling medical management in disasters. AB - The medical aspects of disaster management, also referred to as disaster medicine, is a relatively new medical specialty, the roots of which are to be found in war surgery and traumatology. The main content of disaster medicine is based on empiricism. During the past couple of years, a mathematical approach to some aspects has been added. This may well result in the creation of some order in chaos. This modelling of medical disaster management is important not only in the preparedness phase, but also during the disaster itself and its evaluation. This may in turn result in a decrease in mortality, morbidity and disability amongst disaster casualties. PMID- 10461560 TI - Sympathetic overactivity from fenfluramine-phentermine overdose. AB - A 24-year-old male presented to the emergency department with hyperadrenergic manifestations of fenfluramine-phentermine overdose: tachycardia, mydriasis, fever, diaphoresis, hyperventilation, and combativeness. Sedatives, neuromuscular paralytics, adrenergic antagonists, and mechanical ventilation were required to care for the patient. In addition, the patient had self-inflicted 15% TBSA second degree burns and developed adult respiratory distress syndrome which required continued intubation and mechanical ventilation for 12 days. The patient had split thickness skin grafts for his leg burns on day 11. He was discharged after a 26-day hospital stay. We are unaware of any previously reported cases of fenfluramine-phentermine overdose with such profound degree of sympathetic storm. PMID- 10461561 TI - Emergency therapeutical approach simulating ultrarapid opioid detoxification in methadone withdrawal precipitated by erroneous administration of naltrexone. AB - We report the case of a 30-year-old male, heroin dependent, receiving methadone treatment, who, while staying at home, ingested 50 mg of naltrexone. He immediately developed serious withdrawal symptoms and was admitted to the hospital. In the emergency department the drugs given to counteract the agitation were ineffective, and the patient developed respiratory distress. Anaesthesia with propofol was then started and the patient was intubated, ventilated and hospitalized in the intensive care unit. He was then sedated for 48 hours due to persistent withdrawal signs. When medically stable the patient was transferred to the medical ward where daily treatment with naltrexone and psychological support where started. After 4 days the patient was discharged. Afterwards he did not attend his scheduled outpatient follow-up visits. Treatment with propofol is effective in the case of a patient with a serious withdrawal syndrome secondary to naltrexone overdose during methadone therapy. Despite the actual possibility of getting through the withdrawal symptoms the patient failed to return for follow-up visits, which might be related to a lack of motivation. PMID- 10461562 TI - Hemiplegia hypoglycaemia syndrome. AB - We report the case of a 83-year-old man who presented to the emergency department with hypoglycaemia resembling a cerebrovascular accident. Hypoglycaemic hemiparesis is an under-recognized manifestation of hypoglycaemia. If not recognized and treated promptly, hypoglycaemia may cause irreversible central nervous system injury; it rarely results in death. It is imperative that emergency physicians consider hypoglycaemia in all patients with coma in spite of focal neurological deficit even when the findings seem to be explained initially by other aetiologies. PMID- 10461563 TI - Concurrent administration of amrinone with nitric oxide most improves PaO2/FiO2 in acute respiratory and cardiac failure. AB - A 55-year-old man developed acute respiratory failure, pulmonary hypertension and left heart failure due to acute myocardial infarction. Nitric oxide (NO) inhalation improved arterial oxygenation, decreased pulmonary arterial pressure and increased cardiac output (CO), but combined use of dobutamine with NO produced increases in pulmonary arterial pressure and pulmonary capillary wedge pressure (PCWP). In this patient, amrinone decreased pulmonary arterial pressure and PCWP, and increased PaO2/FiO2 effectively while increasing CO. Combined use of inhaled NO and intravenous amrinone may have beneficial effects for a patient with acute respiratory and cardiac failure. PMID- 10461564 TI - Epileptic disorders with onset in the first year of life: neurological and cognitive outcome. AB - We examined prospectively a series of 150 children with epilepsy beginning in the first year of life. We classified the types of epilepsy into five categories: West syndrome, other epileptic encephalopathies, generalized, partial and undetermined epilepsies. Of 150 patients, 15 died; 135 were followed for at least 4 years. In order to define possible factors influencing prognosis we evaluated neurological and cognitive outcome and made percentage comparisons between groups, for aetiology, age of onset, family history of epilepsy, and psychomotor development before onset. Epileptic encephalopathies as well as the symptomatic forms of West syndrome showed a very poor neurological and cognitive outcome. As previously recognized, only cryptogenic forms of West syndrome had a benign prognosis. For the generalized epilepsies, analysis of different factors, namely late age at onset, cryptogenicity and absence of primary cognitive impairment, indicated a good prognosis. In contrast, partial epilepsies usually had a poor outcome, irrespective of the aetiology, whether cryptogenic or symptomatic. PMID- 10461565 TI - Clinical experiences with topiramate in children with intractable epilepsy. AB - At a tertial referral epilepsy centre 39 children were consecutively enrolled in an open add-on study with topiramate (TPM). All children had intractable epilepsy; the mean seizure frequency was 36 per month, and 31 children were treated with polypharmacy. All but five children were mentally retarded. The initial dose of TPM was 0.5-1 mg/kg daily, slowly titrated with 1-3 mg/kg daily every second week with an estimated target dose of 10 mg/kg daily. At latest follow-up 19 children continued on TPM, three (8%) were seizure-free, eight (21%) had a seizure reduction of more than 50% and eight (21%) improved their general condition. Mean follow-up was 13 months (range 9-36 months). Seizure reduction was seen in focal as well as generalized epilepsies. Adverse effects were reported in 21 cases (54%), weight loss and sedation being most frequent. The mean steady state dose in the children continuing on TPM was at latest follow-up: 14 mg/kg daily (< 5 years), 10 mg/kg daily (5-7 years), 5.8 mg/kg daily (8-17 years). The corresponding plasma level varied from 3 to 45 mumol/litre, and a significant correlation between the daily dose in mg/kg and the plasma level was found. Two patients with progressive myoclonus epilepsy are described separately; one had a dramatic general improvement. It is concluded that TPM seems to be a promising new broad-spectrum anti-epileptic drug, which is efficacious even in epilepsy syndromes, intractable to other new anti-epileptic drugs such as vigabatrin and lamotrigine. PMID- 10461566 TI - Evaluation of hyperhomocysteinaemia in children with stroke. AB - Hyperhomocysteinaemia is associated with an increased risk of arterial vascular disease and thrombosis in adults. Our aim was to study the association of hyperhomocysteinaemia and stroke in children. Since some patients who had suffered a stroke developed seizures and received treatment with anti-epileptic (antifolate) drugs, we also examined the possible interaction between anti epileptic drugs and hyperhomocysteinaemia. Plasma total homocysteine was measured in 68 children with stroke (23 of the 68 were taking anti-epileptic drugs) and 100 children undergoing anti-epileptic treatment but without history of stroke, and we compared the values with our reference values for similar ages (n = 195). Total homocysteine was determined by high profile liquid chromatography with fluorescence detection. Hyperhomocysteinaemia was defined as a homocysteine concentration above the 95th percentile for the reference values. Significant differences were found in total homocysteine values of children with stroke and those taking anti-epileptic drugs compared with our reference values for similar ages, except for the adolescent group. Total homocysteine values above the 95th percentile for the reference values were found in 36% of patients with stroke and 28% of children on anti-epileptic treatment. Total homocysteine concentrations in the 23 patients with both stroke and anti-epileptic drug treatment were similar to those of untreated patients with stroke in all age groups. In summary, systematic screening for hyperhomocysteinaemia should be included in the protocol to investigate the aetiology of stroke, even in paediatrics. Anti-epileptic treatment in children with stroke may be responsible for the mild hyperhomocysteinaemia observed in some of them. A dietary supplement of folate may be of benefit in children with stroke and in patients taking anti-epileptic drugs. PMID- 10461567 TI - Cerebrospinal fluid gangliosides in patients with Rett syndrome and infantile neuronal ceroid lipofuscinosis. AB - Concentrations of the four major brain gangliosides, GM1, GD1a, GD1b and GT1b, biochemical markers of neuronal membranes, were determined in cerebrospinal fluid from a large series of patients with classical Rett syndrome, aged 1.5-21 years at sampling, and from 11 patients with infantile neuronal ceroid lipofuscinosis, aged 1.5-11 years. The results were compared with age-matched healthy controls. Compared with fluid from the control group, the cerebrospinal fluid samples from Rett patients contained significantly reduced levels of gangliosides GD1a and GT1b. In cerebrospinal fluid of the infantile neuronal ceroid lipofuscinosis patients, even the very young ones, all four major brain gangliosides were significantly reduced compared with controls and the concentration levels also differed significantly from those in patients with Rett syndrome. The ganglioside pattern in the brain is reflected in the cerebrospinal fluid early in the course of the disease in Rett syndrome and infantile neuronal ceroid lipofuscinosis. PMID- 10461568 TI - Clinical and brain 18fluoro-2-deoxyglucose positron emission tomographic findings in ethylmalonic aciduria, a progressive neurometabolic disease. AB - We report a 2-year-old boy with ethylmalonic aciduria and vasculopathy syndrome evaluated by 18fluoro-2-deoxyglucose positron emission tomographic (18FDG PET) brain scan, with intense uptake of 18FDG in the caudate nucleus and putamen bilaterally but with no morphological changes on magnetic resonance imaging (MRI). A repeat 18FDG PET brain scan 1 year later showed a significant bilateral decreased uptake of glucose in the putamen and the head of the caudate nucleus as well as a decreased uptake in the frontal lobes. On MRI, there was atrophy and watershed infarcts in the basal ganglia, explaining the loss of glucose uptake. These results reflect a selective vulnerability of the basal ganglia, their functional derangement, and ultimate degeneration. PMID- 10461569 TI - A case of paroxysmal tonic upgaze of childhood with ataxia. AB - Paroxysmal tonic upgaze of childhood is a rare, distinctive, childhood syndrome that may be associated with ataxia and sometimes strabismus or amblyopia. Neurological examination as well as metabolic studies, electroencephalogram and neuroradiological investigations are normal in these patients. Although it has been considered as an age-related, dopa-sensitive dystonia, the exact pathogenetic mechanism is still unknown. Aggravation of attacks by fatigue, intercurrent infection or vaccination, and possible corticomesencephalic dysmaturation may underlie this abnormality. We report on a sporadic case of paroxysmal tonic upgaze with ataxia in which there was prompt aggravation of symptoms with sleep without response to levodopa treatment. This case suggests a different underlying pathogenetic mechanism from dopaminergic pathways for this syndrome. PMID- 10461571 TI - The muscular dystrophies. PMID- 10461570 TI - 'Fou rire prodromique' as the heralding symptom of lenticular infarction, caused by dissection of the internal carotid artery in a 12-year-old boy. AB - A 12-year-old, right-handed boy experienced a pathological fit of laughter before a sudden right hemiplegia. Magnetic resonance imaging showed a left basal ganglia infarction, induced by a left internal carotid dissection. Arteriography revealed an underlying fibromuscular dysplasia. This case study demonstrates that cerebral artery dissection can occur in children and that a basal ganglia infarction may be preceded by pathological laughter called 'fou rire prodromique'. The clinical and anatomical relationship of this paroxysmal event are discussed. PMID- 10461572 TI - Cerebellar astrocytoma and Asperger syndrome. PMID- 10461573 TI - Characterization of color CRT display systems for monochrome applications. AB - Soft-copy presentation of medical images is becoming more and more important as medical imaging is strongly moving toward digital technology, and health care facilities are converting to filmless hospital and radiological information management. Although most medical images are monochrome, frequently they are displayed on color CRTs, particularly if general-purpose workstations or PCs are used for medical viewing. In the present report, general measurement and modeling procedures for the characterization of color CRT monitors for monochrome presentation are introduced. The contributions from the three color channels (red, green, and blue) are weighted according to the spectral sensitivity of the human eye for photopic viewing. The luminance behavior and the resolution capabilities of color CRT monitors are analyzed with the help of photometer and charge-coupled device (CCD) camera measurements. For the evaluation of spatial resolution, a two-dimensional Fourier analysis of special test images containing white noise (broadband response) is employed. A stage model for a color CRT monitor is developed to discuss the effects of scanning and dot sampling. Furthermore, display intrinsic veiling glare and reflectivity of typical color CRT monitors are measured and compared with those of monochrome CRT monitors. The developed methods and models allow one to describe the image quality aspects of color monitors if they are applied for medical monochrome image presentation. Particularly, because of the reduced luminance and dynamic range of color monitors, the calibration and control of their luminance curves is a very important task. For present color CRT monitors, 1,280 x 1,024 turns out to be an intrinsic limit for the displayable matrix of medical images. PMID- 10461574 TI - Aspects of computer security: a primer. AB - As health care organizations continue on the path toward total digital operations, a topic often raised but not clearly understood is that of computer security. The reason for this is simply the vastness of the topic. Computers and networks are complex, and each service offered is a potential security hole. This article describes for the lay person the fundamental points of computer operation, how these can be points attacked, and how these attacks can be foiled- or at least detected. In addition, a taxonomy that should aid system administrators to evaluate and strengthen their systems is described. PMID- 10461575 TI - A client server model to facilitate creation of a medical image teaching library. AB - A simple and convenient system for indexing and archiving medical images used in teaching was developed. The approach was to combine a smart client-side graphical user interface that controlled image size, file format, and keyword structure, and communicated with the hospital information system via hypertext mark-up language, to populate the interface with user selectable pull-down menus. The result is a system that is easily extensible beyond the radiology images for which it was originally designed. Only minor modifications of the client interface are required to adapt the program to accept any file format or image type. PMID- 10461576 TI - A literature review on communication between picture archiving and communication systems and radiology information systems and/or hospital information systems. AB - The purpose of this literature review is to present the concepts surrounding the issue of communication between imaging systems and information systems in radiology and the literature about them. Picture archiving and communication systems (PACS) were developed to combine viewing of modality images, archiving, and distribution of images. When PACS is integrated/interfaced with radiology information systems (RIS) or hospital information systems (HIS), it can merge patient demographics, medical records, and images. To address several issues surrounding communication between PACS and HIS/RIS and to make interface development easier and faster, various organizations have developed standards for the formatting and transfer of clinical data. Additional work continues to better handle these issues. Communication protocol Health Level 7 (HL7) is a standard application protocol used for electronic text data exchange in health care by most HIS/RIS. The imaging communication protocol for PACS is the Digital Imaging and Communications in Medicine (DICOM) standard specification protocol that describes the means of formatting and exchanging images and associated information. PMID- 10461577 TI - Picture archiving and communication systems planning: a methodology. AB - This article presents the Picture Archiving and Communication Systems (PACS) planning methodology used by the Department of Defense's (DOD) Joint Imaging Technology Project Office (JITPO). This methodology evaluates four areas of PACS planning and implementation: strategic planning, clinical scenario planning, installation planning, and implementation planning. The first task is to develop a PACS team, from the local facility, that will execute the program. A written PACS plan is developed by the JITPO, with active input and final say from the site's PACS team. This plan includes the PACS goals and objectives, clinical requirements, facility requirements, and the status of the implementation. This methodology, when applied fully at a military clinical site, has resulted in the site obtaining best "value" in terms of cost and performance by requiring the DOD's contracted PACS vendors to propose a PACS package that meets or exceeds the site's unique requirements. The identification of the requirements and the matching of a known PACS configuration with them has reduced the number of unknowns within the vendors' proposals and created true competition in both initial cost and the cost to maintain PACS in the maintenance years. Although there are certain factors unique to planning a military PACS, such as preselected vendors, the planning methodology described in this article should provide a valuable strategy for any hospital planning a PACS. PMID- 10461578 TI - Evaluation: beyond the rhetoric. PMID- 10461579 TI - Achievable benchmarks of care: the ABCs of benchmarking. AB - Benchmarking is generally considered to be an important tool for quality improvement. Traditional approaches to benchmarking have relied on subjective identification of 'leaders in the field'. We derive an objective, reproducible and attainable Achievable Benchmark of Care (ABC) by measuring and analysing performance on process-of-care indicators. Three characteristics of the ABC that we deem essential are: (1) benchmarks represent a measurable level of excellence; (2) benchmarks are demonstrably attainable; (3) benchmarks are derived from data in an objective, reproducible and predetermined fashion. From these characteristics it follows that (4) providers with high performance are selected to define a level of excellence in a predetermined fashion, but (5) providers with high performance on small numbers of cases do not influence unduly benchmark levels. We use the 'pared mean' to operationalize the ABC. Roughly, the pared mean summarizes the performance of top-ranked providers whereby at least 10% of the patient pool across all providers is included. Bayesian estimators for adjustment of performance of providers with small sample sizes are used to rank providers. Randomized controlled trials to assess the independent effect of the ABC in quality improvement projects are under way. We have developed a methodology objectively and reproducibly to derive a level of excellent, attainable performance, based on measured performance by a group of providers. The ABC can be applied to groups of providers in communities, to institutions and departments within them, or to individual practitioners. PMID- 10461580 TI - Systematic validation of disease models for pharmacoeconomic evaluations. Swiss HIV Cohort Study. AB - Pharmacoeconomic evaluations are often based on computer models which simulate the course of disease with and without medical interventions. The purpose of this study is to propose and illustrate a rigorous approach for validating such disease models. For illustrative purposes, we applied this approach to a computer based model we developed to mimic the history of HIV-infected subjects at the greatest risk for Mycobacterium avium complex (MAC) infection in Switzerland. The drugs included as a prophylactic intervention against MAC infection were azithromycin and clarithromycin. We used a homogenous Markov chain to describe the progression of an HIV-infected patient through six MAC-free states, one MAC state, and death. Probability estimates were extracted from the Swiss HIV Cohort Study database (1993-95) and randomized controlled trials. The model was validated testing for (1) technical validity (2) predictive validity (3) face validity and (4) modelling process validity. Sensitivity analysis and independent model implementation in DATA (PPS) and self-written Fortran 90 code (BAC) assured technical validity. Agreement between modelled and observed MAC incidence confirmed predictive validity. Modelled MAC prophylaxis at different starting conditions affirmed face validity. Published articles by other authors supported modelling process validity. The proposed validation procedure is a useful approach to improve the validity of the model. PMID- 10461581 TI - Mandatory practice self-appraisal: moving towards outcomes based continuing education. AB - Most organized physician continuing educational activities are undertaken without assessing their effects on long-term changes in physician practice patterns, patient outcomes or return on investment. Practice audit and practice self appraisal are two activities that can be used to achieve these objectives. It is recommended that these be promoted from an educational perspective. A series of principles, including the mandatory application of practice audit and practice self-appraisal are proposed to guide the process. The identification of issues relating to learning, diffusion of information and behavioural change required to facilitate this change are briefly discussed. PMID- 10461582 TI - The role of audit in making do not resuscitate decisions. AB - Audit has been proposed as a useful means of implementing and evaluating 'Do Not Resuscitate (DNR)' policies. This paper explores the background to the introduction of DNR policies and reviews published reports of their use in clinical practice. Problems with auditing and implementing DNR policies are highlighted. PMID- 10461583 TI - Audit in British general practice: domination or disillusionment? AB - The history of audit is traced over the past 30 years demonstrating how it has come to dominate attempts to improve the quality of patient care in British general practice. Nevertheless it is still uncertain whether audit usually or occasionally yields such improvements. There is much more to learn about audit including whether any benefits it brings are through professional education. I suggest that the use of audit as a foundation upon which clinical governance is built is not a secure one. PMID- 10461584 TI - The lack of integration of clinical audit and the maintenance of medical dominance within British hospital trusts. AB - Concerns have been expressed repeatedly about the effectiveness of clinical audit. Some have argued that this is limited by the lack of integration within day-to-day practice and with other NHS policy initiatives. We aimed to explore what mechanisms were being used to develop annual clinical audit programmes within NHS Trusts, and to describe the influence of other initiatives on this; to understand how such influences are exerted; and to understand the role of key players, in order to inform future programme development. Semi-structured face-to face interviews were performed with Chairs of Clinical Audit Committees, Clinical Audit Managers and Co-ordinators (N = 15) in the former Yorkshire Region of the NHS in England. Concerns about the development, planning and integration of clinical audit focused upon an almost exclusive medical dominance and upon how audit leadership could be delivered within the context of hospital management structures. The lack of an overall plan for the development of clinical audit in most sites was seen as enabling the doctors' agenda to dominate. Purchasing authorities were recognized as being important, but often with limited influence. Other influences on the audit agenda, such as research and development (R&D) and clinical risk management, were rarely well co-ordinated. These findings concur with previous studies in identifying a wide range of constraints on the progress of audit. Several of these constraints operate within the internal environment, for example the doctors' agenda, and concerns about management involvement. Such constraints require resolution in order to facilitate the integration of audit with other initiatives and to achieve the goals of audit effectively. Clinical effectiveness and clinical governance may offer a means of facilitating this integration. PMID- 10461585 TI - Public trust and accountability for clinical performance: lessons from the national press reportage of the Bristol hearing. AB - The General Medical Council hearing into events at the Bristol Royal Infirmary resulted in verdicts of serious professional misconduct against three senior doctors. After the longest-running hearing in the GMC's history the press response was fierce. This paper reviews the reporting of the Bristol case (and issues arising from it) in the main broadsheet and tabloid national newspapers (dailies and Sundays) in the 5-week period around the GMC's delivery of the verdicts and subsequent sentencing. The aim was to describe the main themes emerging from the press coverage and to assess the implications for future debates over clinical performance and accountability. Media interest in the Bristol case was intense (184 published items in 5 weeks). The reporting was emotive and largely hostile, raising doubts about not just isolated lapses of care but also the possibility of more systematic failings. Diminished trust and reduced public confidence were recurrent themes, powerfully expressed. Professional self-regulation received scathing criticism, with calls for more public access to individual performance data. Future debates about clinical governance will need to take account of the new public context in the wake of Bristol. Arguments about the relative merits of self-regulation or data-driven performance management systems now need to be played out for a knowing and openly sceptical print media. PMID- 10461586 TI - The pen is mightier than the scalpel. Commentary on the paper--public trust, and accountability for clinical performance: lessons from the national press reporting of the Bristol hearing. PMID- 10461588 TI - What are quality of life improvements measuring and do they last? An explanatory model. PMID- 10461587 TI - A consultant looks at the NHS today. PMID- 10461589 TI - Research into emergency treatments--could the offer of 'advance directives' help? PMID- 10461590 TI - Bioethics in and from Asia. PMID- 10461591 TI - Family consent, communication, and advance directives for cancer disclosure: a Japanese case and discussion. AB - The dilemma of whether and how to disclose a diagnosis of cancer or of any other terminal illness continues to be a subject of worldwide interest. We present the case of a 62-year-old Japanese woman afflicted with advanced gall bladder cancer who had previously expressed a preference not to be told a diagnosis of cancer. The treating physician revealed the diagnosis to the family first, and then told the patient: "You don't have any cancer yet, but if we don't treat you, it will progress to a cancer". In our analysis, we examine the role of family consent, communication patterns (including ambiguous disclosure), and advance directives for cancer disclosure in Japan. Finally, we explore the implications for Edmund Pellegrino's proposal of "something close to autonomy" as a universal good. PMID- 10461592 TI - Survey of Japanese physicians' attitudes towards the care of adult patients in persistent vegetative state. AB - OBJECTIVES: Ethical issues have recently been raised regarding the appropriate care of patients in persistent vegetative state (PVS) in Japan. The purpose of our study is to study the attitudes and beliefs of Japanese physicians who have experience caring for patients in PVS. DESIGN AND SETTING: A postal questionnaire was sent to all 317 representative members of the Japan Society of Apoplexy working at university hospitals or designated teaching hospitals by the Ministry of Health and Welfare. The questionnaire asked subjects what they would recommend for three hypothetical vignettes that varied with respect to a PVS patient's previous wishes and the wishes of the family. RESULTS: The response rate was 65%. In the case of a PVS patient who had no previous expressed wishes and no family, 3% of the respondents would withdraw artificial nutrition and hydration (ANH) when the patient did not require any other life-sustaining treatments, 4% would discontinue ANH, and 30% would withhold antibiotics when the patient developed pneumonia. Significantly more respondents (17%) would withdraw ANH in the case of a PVS patient whose previous wishes and family agreed that all life support be discontinued. Most respondents thought that a patient's written advance directives would influence their decisions. Forty per cent of the respondents would want to have ANH stopped and 31% would not want antibiotics administered if they were in PVS. CONCLUSIONS: Japanese physicians tend not to withdraw ANH from PVS patients. Patients' written advance directives, however, would affect their decisions. PMID- 10461593 TI - The status of the do-not-resuscitate order in Chinese clinical trial patients in a cancer centre. AB - OBJECTIVE: To report and analyse the pattern of end-of-life decision making for terminal Chinese cancer patients. DESIGN: Retrospective descriptive study. SETTING: A cancer clinical trials unit in a large teaching hospital. PATIENTS: From April 1992 to August 1997, 177 consecutive deaths of cancer clinical trial patients were studied. MAIN MEASUREMENT: Basic demographic data, patient status at the time of signing a DNR consent, or at the moment of returning home to die are documented, and circumstances surrounding these events evaluated. RESULTS: DNR orders were written for 64.4% of patients. Patients in pain (odds ratio 0.45, 95% CI 0.22-0.89), especially if requiring opioid analgesia (odds ratio 0.40, 95% CI 0.21-0.77), were factors associated with a higher probability of such an order. Thirty-five patients were taken home to die, a more likely occurrence if the patient was over 75 years (odds ratio 0.12, 95% CI 0.04-0.34), had children (odds ratio 0.14, 95% CI 0.02-0.79), had Taiwanese as a first language (odds ratio 6.74, 95% CI 3.04-14.93), or was unable to intake orally (odds ratio 2.73, 95% CI 1.26-5.92). CPR was performed in 30 patients, none survived to discharge. CONCLUSIONS: DNR orders are instituted in a large proportion of dying Chinese cancer patients in a cancer centre, however, the order is seldom signed by the patient personally. This study also illustrates that as many as 20% of dying patients are taken home to die, in accordance with local custom. PMID- 10461594 TI - Ancient Chinese medical ethics and the four principles of biomedical ethics. AB - The four principles approach to biomedical ethics (4PBE) has, since the 1970s, been increasingly developed as a universal bioethics method. Despite its wide acceptance and popularity, the 4PBE has received many challenges to its cross cultural plausibility. This paper first specifies the principles and characteristics of ancient Chinese medical ethics (ACME), then makes a comparison between ACME and the 4PBE with a view to testing out the 4PBE's cross-cultural plausibility when applied to one particular but very extensive and prominent cultural context. The result shows that the concepts of respect for autonomy, non maleficence, beneficence and justice are clearly identifiable in ACME. Yet, being influenced by certain socio-cultural factors, those applying the 4PBE in Chinese society may tend to adopt a "beneficence-oriented", rather than an "autonomy oriented" approach, which, in general, is dissimilar to the practice of contemporary Western bioethics, where "autonomy often triumphs". PMID- 10461595 TI - The ambiguity about death in Japan: an ethical implication for organ procurement. AB - In the latter half of the twentieth century, developed countries of the world have made tremendous strides in organ donation and transplantation. However, in this area of medicine, Japan has been slow to follow. Japanese ethics, deeply rooted in religion and tradition, have affected their outlook on life and death. Because the Japanese have only recently started to acknowledge the concept of brain death, transplantation of major organs has been hindered in that country. Currently, there is a dual definition of death in Japan, intended to satisfy both sides of the issue. This interesting paradox, which still stands to be fully resolved, illustrates the contentious conflict between medical ethics and medical progress in Japan. PMID- 10461597 TI - Should childhood immunisation be compulsory? AB - Immunisation is offered to all age groups in the UK, but is mainly given to infants and school-age children. Such immunisation is not compulsory, in contrast to other countries, such as the United States. Levels of immunisation are generally very high in the UK, but the rates of immunisation vary with the public perception of the risk of side effects. This article discusses whether compulsory vaccination is acceptable by considering individual cases where parents have failed to give consent or have explicitly refused consent for their children to be immunised. In particular, the rights of: a parent to rear his/her child according to his/her own standards; the child to receive health care, and the community to be protected from vaccine-preventable infectious disease are considered. The conclusion of the article is that compulsory vaccination cannot, with very few exceptions, be justified in the UK, in view of the high levels of population immunity which currently exist. PMID- 10461596 TI - Should Zelen pre-randomised consent designs be used in some neonatal trials? AB - My aim is to suggest that there is a case for using a randomised consent design in some neonatal trials. As an example I use the trials of extracorporeal membrane oxygenation (ECMO) in neonates suffering pulmonary hypertension. In some trials the process of obtaining consent has the potential to harm the subject, for example, by disappointing those who end in the control group and by creating additional anxiety at times of acute illness. An example of such were the trials of extracorporeal membrane oxygenation (ECMO) in neonates suffering pulmonary hypertension. Pre-randomised consent could avoid or lessen these harms. However, a number of ethical objections are made to these research designs. They involve denial of information, using people, denial of choice, and "overselling" of allocated treatment. Furthermore, they are the wrong response; better communication might be the answer, for example. I argue that these objections are not completely persuasive. However, they are enough to suggest caution in the use of such designs. PMID- 10461598 TI - Genetic medicine: an experiment in community-expert interaction. AB - This project tested a two-way model of communication between lay groups and experts about genetic medicine in Perth, Western Australia. Focus group discussion with community group participants was followed by a communication workshop between community group participants and experts. Four groups of concerns or themes emerged from discussion: clinical considerations; legislative concerns; research priorities, and ethical and wider considerations. Community group concerns are not always met by the actions of "experts". This is, in part, because of the differing life-worlds of each group. However, the communication workshop showed the potential of two-way communication for both lay and expert members in understanding the others' viewpoint. Further, the approach developed here offers one possible way for community groups to participate in a substantial way in policy formulation processes. PMID- 10461599 TI - In defence of medical ethics. AB - A number of recent publications by the philosopher David Seedhouse are discussed. Although medicine is an eminently ethical enterprise, the technical and ethical aspects of health care practices can be distinguished, therefore justifying the existence of medical ethics and its teaching as a specific part of every medical curriculum. The goal of teaching medical ethics is to make health care practitioners aware of the essential ethical aspects of their work. Furthermore, the contention that rational bioethics is a fruitless enterprise because it analyses non-rational social events seems neither theoretically tenable nor to be borne out by actual practice. Medical ethics in particular and bioethics in general, constitute a field of expertise that must make itself understandable and convincing to relevant audiences in health care. PMID- 10461600 TI - Camouflage is no defence--a response to Kottow. AB - The author responds to Professor Kottow's criticisms, explaining numerous errors and misconceptions. PMID- 10461601 TI - Voluntary euthanasia in The Netherlands. PMID- 10461602 TI - Life support or molecular maintenance. PMID- 10461603 TI - The interactive topography of the future. PMID- 10461604 TI - Agreement and disagreement between "metabolic viability" and "contractile reserve" in akinetic myocardium. AB - BACKGROUND: In patients with chronic coronary artery disease and depressed left ventricular function, assessment of residual viability in akinetic myocardium is important for therapeutic management. Intact perfusion, preserved metabolism, and presence of contractile reserve are different aspects of cellular viability. However, not all viable cells exhibit all characteristics; it is thought that contractile reserve is less often preserved compared with metabolic activity or intact perfusion. In this study we performed a direct comparison between perfusion imaging with thallium-201 single photon emission computed tomography (SPECT), metabolic imaging with F18-fluorodeoxyglucose SPECT, and assessment of contractile reserve with low-dose dobutamine echocardiography in akinetic myocardium. METHODS AND RESULTS: Forty patients with depressed left ventricular function (mean left ventricular ejection fraction 31% +/- 16%) were studied. Resting echocardiography showed akinesis in 165 (32%) segments. Most (n = 154, 93%) of these segments demonstrated resting hypoperfusion. F18-fluorodeoxyglucose imaging revealed a perfusion-metabolism mismatch in 41 segments and a match in 113 segments. Contractile reserve was present in 33 (80%) of the segments with a perfusion-metabolism mismatch and in 7 (6%) segments with a match (P < .0005). Of the 11 segments with normal perfusion, only 5 (45%) showed contractile reserve. The agreement between SPECT and dobutamine echocardiography was 87%. Although 94% of the segments that were nonviable on scintigraphy did not show contractile reserve, the disagreement between SPECT and dobutamine echocardiography was caused mainly by the absence of contractile reserve in 27% of the segments that were viable on scintigraphy. CONCLUSION: This study shows a good agreement between SPECT and dobutamine echocardiography, although a substantial number of segments with preserved viability on SPECT do not exhibit contractile reserve, indicating underestimation of viability by dobutamine echocardiography compared with F18-fluorodeoxyglucose imaging. PMID- 10461605 TI - Comparison of exercise, dipyridamole, adenosine, and dobutamine stress with the use of Tc-99m tetrofosmin tomographic imaging. AB - PURPOSE: The purpose of this study was to compare defect extent and severity and myocardial uptake with exercise and pharmacologic stress with technetium-99m (Tc 99m) tetrofosmin tomographic myocardial perfusion imaging. BACKGROUND: Detection of stress-induced myocardial perfusion defects depends on both a disparity in blood flow between normal and stenotic vessels and the extraction fraction and linearity of myocardial uptake of the tracer. There are limited clinical data for exercise or pharmacologic stress with Tc-99m tetrofosmin tomographic myocardial perfusion imaging. METHODS: Thirty-one patients with coronary artery disease and 7 with a < 5% likelihood of coronary artery disease underwent on separate days Tc 99m tetrofosmin single-photon emission computed tomographic imaging at rest and after exercise, dipyridamole, adenosine, and dobutamine stress. Images were interpreted by a blinded consensus of 3 experienced readers with a 17-segment model and 5-point scoring system. RESULTS: Compared with exercise, the summed stress score was smaller with dipyridamole (P < .01), and the reversibility score was smaller with both dipyridamole (P < .01) and dobutamine (P < .05), whereas the number of abnormal and reversible segments was less with both dipyridamole (P < .01 and P < .001, respectively) and dobutamine (both P < .05). No significant differences were found in the summed stress or reversibility scores and the number of abnormal or reversible segments between exercise and adenosine. CONCLUSIONS: Compared with exercise, defect extent, severity, and reversibility are less with dipyridamole and dobutamine with Tc-99m tetrofosmin single photon emission computed tomographic imaging. PMID- 10461606 TI - Transient left ventricular dilation at quantitative stress-rest sestamibi tomography: clinical, electrocardiographic, and angiographic correlates. AB - BACKGROUND: Few data are available regarding the incidence and significance of transient left ventricular (LV) dilation on stress sestamibi single photon emission computed tomography (SPECT), which is different from thallium-201 studies because images are acquired late after tracer injection. METHODS: We studied 234 patients with ischemic heart disease and interpretable electrocardiograms undergoing stress-rest sestamibi SPECT on separate days. Sestamibi uptake defect extent was quantified on SPECT polar maps. Epicardial and endocardial transient dilation indexes (TDI) were also calculated. RESULTS: According to our normal TDI values, 148 patients (63%) had no dilation and 86 patients (37%) had abnormal endocardial TDI; a global LV dilation (abnormal endocardial and epicardial TDI) was observed in 19 patients (8%). ST-segment depression was more frequent in patients with transient LV dilation (55%) than in those without (36%; P < .01), as were the extent of stress hypoperfusion (13% +/- 12% vs 6% +/- 7% in patients with no dilation; P < .001) and the angiographic severity score (11.4 +/- 5.9 vs 9.2 +/- 3.7; P < .05). At multivariate analysis, stress hypoperfusion was the sole predictor of transient LV dilation. CONCLUSIONS: Transient LV cavity dilation is frequent on stress sestamibi SPECT. Ventricular cavity dilation is more common than global dilation and suggests subendocardial ischemia. It is related to a greater amount of jeopardized myocardium and is strongly associated with electro-cardiographic signs of ischemia. PMID- 10461607 TI - Clinical implication of down-scatter in attenuation-corrected myocardial SPECT. AB - BACKGROUND: Interpretation of myocardial perfusion single photon emission computed tomography (SPECT) studies is hampered by attenuation artifacts. Attenuation correction methods with simultaneous emission and transmission are now commercially available. However, it has been observed in clinical practice that attenuation correction without down-scatter correction in a 1-day rest/stress myocardial perfusion protocol may lead to serious interpretation errors. Therefore the aim of this study was to study errors resulting from down scatter under realistic conditions, thus providing a background for the assessment of further corrections. METHODS AND RESULTS: Forty-six patients underwent myocardial perfusion scintigraphy in a 1-day technetium 99m-tetrofosmin rest-stress SPECT protocol, with a moving 153Gd line-source device for attenuation correction without down-scatter correction. Short-axis slices were quantified as inferior/anterior, septal/lateral, and apical/remainder count ratios. The changes at rest (350 MBq) and exercise (900 MBq) induced by attenuation correction were studied. Attenuation correction gave differences in apparent perfusion between rest and exercise not seen before correction. The gender differences in inferior-anterior ratio were greatly reduced after correction at rest but remained at exercise. A torso phantom study indicated that these results were due to under-correction at exercise because of down-scatter. CONCLUSIONS: Down-scatter results in an underestimation of attenuation in simultaneous emission and transmission, if not accurately accounted for. Particularly, a high-dose study compared with a low-dose study, as in the 1-day protocol, might cause serious interpretation errors. PMID- 10461609 TI - Comparative performance of gated perfusion SPECT wall thickening, delayed thallium uptake, and F-18 fluorodeoxyglucose SPECT in detecting myocardial viability. AB - To evaluate the comparative abilities of gated single photon emission computed tomography (SPECT) wall thickening, delayed thallium-201 (Tl-201) SPECT, and F-18 fluorodeoxyglucose (FDG) SPECT in detecting myocardial viability, 23 patients with previous myocardial infarction and clinically suspected viability were studied. Each patient had at least 1 extensive fixed perfusion defect on rest/stress technetium-99m sestamibi SPECT. A total of 41 major vascular territories had fixed defects. The mean (+/- 1 SD) left ventricular ejection fraction determined from gated perfusion SPECT was 26% +/- 11%. Wall thickening was assessed in a semiquantitative fashion by the regional increase in myocardial intensity during systole and was considered normal when a > or = 20% increase was observed. Tl-201 SPECT was acquired 4 hours after resting tracer injection was administered. Viability was considered present when regional defect Tl-201 count density, determined by quantitative analysis, was > 20% greater than that on the resting sestamibi scan. FDG SPECT was performed independently with a 10 mCi F-18 FDG dose after oral glucose loading was performed. A camera equipped with ultrahigh energy collimation was used. Quantitative criteria for viability were the same as for Tl-201. In the 23 patients viability within the fixed sestamibi defects was manifest by preserved wall thickening in 8 patients, delayed Tl-201 uptake in 10 patients, and FDG uptake in 18 patients. Nine major vascular territories with fixed defects were judged viable by wall thickening, 11 by Tl 201 SPECT, and 24 by FDG SPECT (P = .0009). We conclude that FDG SPECT demonstrates more evidence of myocardial viability than either gated sestamibi wall thickening or delayed Tl-201 SPECT. PMID- 10461608 TI - Accuracy of left ventricular ejection fraction determined by gated myocardial perfusion SPECT with Tl-201 and Tc-99m sestamibi: comparison with first-pass radionuclide angiography. AB - BACKGROUND: We compared estimates of left ventricular ejection fraction (LVEF) assessed by gated single photon emission computed tomography (SPECT), using both technetium-99m sestamibi and thallium-201, with those obtained by first-pass radionuclide angiography (FPRNA) in patients with a broad spectrum of LVEF and perfusion abnormalities. METHODS: Sixty-three patients were randomly selected to undergo a dual isotope gated SPECT study (rest Tl-201 followed by adenosine Tc 99m sestamibi scintigraphy). Studies were processed by use of the Cedars quantitative gated SPECT software. FPRNA was acquired during an intravenous bolus injection of Tc-99m sestamibi and processed with a commercially available software. RESULTS: The estimates of LVEF were similar (P = NS) with Tl-201 gated SPECT (54% +/- 15%), Tc-99m gated SPECT (54% +/- 16%), and FPRNA (54% +/- 12%). There was an excellent correlation between Tc-99m and Tl-201 gated SPECT (Pearson's r = 0.92, P < .0001). There were also good linear correlations between Tc-99m sestamibi gated SPECT and FPRNA (Pearson's r = 0.85, P < .0001), as well as between Tl-201 gated SPECT and FPRNA (Pearson's r = 0.84, P < .0001). In the 16 patients with LVEF < 50%, Tc-99m sestamibi gated SPECT and FPRNA (Pearson's r = 0.84, P < .0001) and Tl-201 gated SPECT and FPRNA (Pearson's r = 0.92, P < .0001) correlated well. CONCLUSION: LVEF can be accurately assessed by gated SPECT with either Tc-99m sestamibi or Tl-201 in properly selected patients with normal or depressed left ventricular function. PMID- 10461610 TI - Cardiac gated equilibrium radionuclide angiography and multiharmonic Fourier phase analysis: optimal acquisition parameters in arrhythmogenic right ventricular cardiomyopathy. AB - BACKGROUND: Multiharmonic Fourier phase analysis of radionuclide angiography is a well-established method for the diagnosis of arrhythmogenic right ventricular cardiomyopathy. We sought to determine the optimal acquisition parameters: number of frames per cycle and number of counts per frame, with all other acquisition and processing parameters being fixed. METHODS AND RESULTS: Radionuclide angiography with list mode acquisition was performed in 10 normal subjects (pilot group) and 11 patients with arrhythmogenic right ventricular cardiomyopathy (validation group), allowing the reconstruction of electrocardiography-gated constant phase studies with different parameters: 16, 24, and 32 frames per cycle and 200, 400, 600, and 800 kcounts per frame. Three harmonics Fourier phase analysis was applied, and optimal acquisition parameters (defined as those providing best homogeneous phase distribution histogram in the pilot group) were defined as judged by the H3 right ventricular phase SD and delta 95%. These were 16 frames per cycle and 600 kcounts per frame. Then we verified in the validation group that these optimal acquisition parameters did not induce any significant relative loss of information compared with other acquisition parameters with more temporal resolution (24 and 32 frames per cycle) or more statistics (800 kcounts per frame). This result was realized by the calculation of normalized H3 right ventricular SD, right ventricular delta 95%, and (SD[left ventricle] - SD[right ventricle]). CONCLUSIONS: In practice, 16 frames per cycle and 600 kcounts per frame are optimal for multiharmonic Fourier phase analysis, with all other acquisition and processing variables being fixed as specified. PMID- 10461611 TI - Radionuclide imaging probes for expressed proteins. PMID- 10461612 TI - An introduction to attenuation correction. AB - Attenuation correction techniques have demonstrated improved diagnostic accuracy and quality of myocardial perfusion SPECT images in limited studies. The future success of these methods relies largely on understanding the characteristics of the images and their interpretative meaning, as well as their limitations. It will be important to define the impact on patient management decisions, quantitation, laboratory efficiency physician confidence, and communication of important findings. Advances in these areas will help position nuclear cardiology to compete with other imaging modalities. As this technology matures, the technologist has an essential role in ensuring efficient use of these techniques and maximizing the quality of this promising new way to image patients. PMID- 10461613 TI - Metabolic imaging and contractile reserve for assessment of myocardial viability: friends or foes? PMID- 10461614 TI - Current clinical relevance of cardiovascular magnetic resonance and its relationship to nuclear cardiology. PMID- 10461615 TI - American Society of Nuclear Cardiology position statement on electrocardiographic gating of myocardial perfusion SPECT scintigrams. PMID- 10461616 TI - Dipyridamole-Tc-99m-tetrofosmin SPECT imaging can identify provokable ischemia in heart transplantation. PMID- 10461617 TI - The patient who assumed his analyst's identity. PMID- 10461618 TI - Repetition of the mother-infant dyad and the process of dying in the psychotherapy of an HIV-positive man. PMID- 10461619 TI - The relationship of repression to reality testing in adult women who report childhood sexual and physical abuse. PMID- 10461620 TI - Epigenesis and sexual orientation: a report of five bisexual males. PMID- 10461621 TI - Dynamic psychotherapy and the disabled psychiatrist. PMID- 10461622 TI - Unresolved problems in the dream of Irma's Injection. PMID- 10461623 TI - Mask and steel: Mishima--when life imitates art. PMID- 10461624 TI - Medieval antecedents of the therapeutic alliance. PMID- 10461625 TI - Is there a future in disillusion? Constructionist and deconstructionist approaches in psychoanalysis. PMID- 10461626 TI - HSUS withdraws FDA lawsuit. PMID- 10461627 TI - The two sides on hormone-treated beef. PMID- 10461628 TI - What is your diagnosis? Pulmonary bullous emphysema with pneumothorax and subcutaneous emphysema. PMID- 10461629 TI - Geographic distribution of female and male veterinarians in the United States, 1998. PMID- 10461630 TI - Cardiopulmonary measurements in dogs undergoing gastropexy without gastrectomy for correction of gastric dilatation-volvulus. AB - OBJECTIVE: To measure cardiopulmonary variables, including cardiac index, in dogs with naturally acquired gastric dilatation-volvulus (GDV). DESIGN: Prospective clinical study. ANIMALS: 6 dogs with GDV. PROCEDURE: In addition to typical medical and surgical management of GDV, the dorsal metatarsal and pulmonary arteries and right atrium of the dogs were catheterized to obtain cardiopulmonary measurements before and during anesthesia and surgery. RESULTS: All dogs underwent gastropexy but none required gastrectomy. Mean cardiac index and mean arterial blood pressure for this small population of dogs with GDV were not significantly different from those reported for clinically normal awake or anesthetized dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Dogs with naturally acquired GDV without gastric necrosis may not have the classic characteristics, including decreased cardiac index and hypotension, of hypovolemic circulatory shock. PMID- 10461631 TI - Effects of phenobarbital treatment on serum thyroxine and thyroid-stimulating hormone concentrations in epileptic dogs. AB - OBJECTIVE: To determine whether phenobarbital treatment of epileptic dogs alters serum thyroxine (T4) and thyroid-stimulating hormone (TSH) concentrations. DESIGN: Cross-sectional study. ANIMALS: 78 epileptic dogs receiving phenobarbital (group 1) and 48 untreated epileptic dogs (group 2). PROCEDURE: Serum biochemical analyses, including T4 and TSH concentrations, were performed for all dogs. Additional in vitro analyses were performed on serum from healthy dogs to determine whether phenobarbital in serum interferes with T4 assays or alters free T4 (fT4) concentrations. RESULTS: Mean serum T4 concentration was significantly lower, and mean serum TSH concentration significantly higher, in dogs in group 1, compared with those in group 2. Thirty-one (40%) dogs in group 1 had serum T4 concentrations less than the reference range, compared with 4 (8%) dogs in group 2. All dogs in group 2 with low serum T4 concentrations had recently had seizure activity. Five (7%) dogs in group 1, but none of the dogs in group 2, had serum TSH concentrations greater than the reference range. Associations were not detected between serum T4 concentration and TSH concentration, age, phenobarbital dosage, duration of treatment, serum phenobarbital concentration, or degree of seizure control. Signs of overt hypothyroidism were not evident in dogs with low T4 concentrations. Addition of phenobarbital in vitro to serum did not affect determination of T4 concentration and only minimally affected fT4 concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Clinicians should be aware of the potential for phenobarbital treatment to decrease serum T4 and increase TSH concentrations and should use caution when interpreting results of thyroid tests in dogs receiving phenobarbital. PMID- 10461632 TI - Administration of diminazene aceturate or imidocarb dipropionate for treatment of cytauxzoonosis in cats. AB - Bobcats (Lynx rufus) are the reservoir hosts for Cytauxzoon felis, the causative agent of cytauxzoonosis. Cytauxzoonosis is a highly fatal tickborne blood protozoal disease of domestic and exotic cats. Treatment of clinically affected cats has generally been unrewarding. In our report, 6 of 7 cats had signs of illness and laboratory findings indicative of cytauxzoonosis and were successfully treated with 2 i.m. injections of diminazene aceturate or imidocarb dipropionate (2 mg/kg [0.9 mg/lb] of body weight, each). One cat died after the first injection of diminazene. Additional treatment with isotonic fluids i.v. and heparin s.c. were used as supportive care for dehydration and disseminated intravascular coagulation that were detected by laboratory testing between diminazene or imidocarb treatments. Atropine was effective in recovery and preventing adverse reactions associated with imidocarb treatment of 1 cat. PMID- 10461633 TI - Use of cyproheptadine to control urine spraying in a castrated male domestic cat. AB - A 10-year-old castrated male domestic cat was admitted to the hospital because of lifelong urine spraying of vertical surfaces. A diagnosis of territorial urine marking was made. Laboratory analytes for urine analysis, hemogram, serum biochemical analysis, and serum thyroxine concentration were within reference ranges, and testosterone concentration was consistent with the reference range of castrated male cats. Treatment included behavior modification and the administration of cyproheptadine, which resulted in the immediate arrest of undesirable urine marking. Cyproheptadine administration was adjusted to determine the lowest dosage that effectively maintained the cat's consistent use of the litter box. Cyproheptadine administration was recommended for at least 1 year before any attempt to withdraw its use. A follow-up phone call to the owner 8 months after the beginning of treatment revealed that the cat continued to have remission of inappropriate urination. Cyproheptadine, an antihistamine prescribed for its orexigenic effects in cats, has antiandrogenic effects in other species. Information in this report indicates that cyproheptadine is effective in the control of urine spraying even in castrated cats. PMID- 10461634 TI - Drug disposition and dosage determination of once daily administration of gentamicin sulfate in horses after abdominal surgery. AB - OBJECTIVE: To evaluate pharmacokinetics of once daily i.v. administration of gentamicin sulfate to adult horses that had abdominal surgery. DESIGN: Prospective study. ANIMALS: 28 adult horses that underwent abdominal surgery for colic. PROCEDURE: 14 horses were treated with each dosage of gentamicin (i.e., 6.6 or 4 mg/kg, i.v., q 24 h) and blood samples were collected for pharmacokinetic analysis. Plasma gentamicin concentrations were measured by use of a fluorescence polarization immunoassay. Pharmacokinetic analysis measured the elimination half-life, volume of distribution, and gentamicin total systemic clearance. Treatment outcome, CBC, and serum creatinine concentrations were recorded. RESULTS: 1 horse in the high-dosage group died. All other horses successfully recovered, and did not develop bacterial infection or have evidence of drug toxicosis resulting in renal injury. Mean pharmacokinetic variables for gentamicin administration at a high or low dosage (i.e., 6.6 or 4 mg/kg, i.v., q 24 h) were half-life of 1.47 and 1.61 hours, volume of distribution of 0.17 and 0.17 L/kg, and systemic clearance of 1.27 and 1.2 ml/kg/min, respectively. Mean serum creatinine concentration was 1.74 and 1.71 for the high and low dosages, respectively, and serum creatinine concentration was not correlated with gentamicin clearance. CONCLUSIONS AND CLINICAL RELEVANCE: Gentamicin administration at a dosage of 4 mg/kg, i.v., every 24 hours, will result in plasma concentrations that are adequate against susceptible bacteria with a minimum inhibitory concentration (MIC) of < or = 2.0 micrograms/ml. Gentamicin administration at a calculated dosage of 6.8 mg/kg, i.v., every 24 hours will result in optimum plasma concentrations against susceptible bacteria with a MIC of < or = 4.0 micrograms/ml. PMID- 10461635 TI - Isolation of Salmonella organisms from the mesenteric lymph nodes of horses at necropsy. AB - OBJECTIVES: To determine the prevalence of Salmonella infections in horses at necropsy. DESIGN: Cross-sectional prevalence survey. ANIMALS: 102 horses. PROCEDURE: Mesenteric lymph nodes were collected from horses that were necropsied. Horses had died or were euthanatized because of severe disease or at the request of the owner. Twenty-eight of the horses were racehorses euthantized following acute catastrophic injuries on the racetrack. Mesenteric lymph nodes were submitted for Salmonella culture via direct plating of tissue specimens on MacConkey agar and by use of 4 enrichment culture techniques that used tetrathionate and selenite enrichment broth and brilliant green and Salmonella Shigella selective plating media. RESULTS: Salmonella typhimurium was isolated from the mesenteric lymph nodes of 2 foals (2/102, 1.96% of the horses). Salmonella organisms were not isolated from the mesenteric lymph nodes of adult horses. CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of Salmonella infections in horses of our study (1.96%) suggests that the results of cross-sectional surveys, using bacteriologic culture to determine prevalence of Salmonella infection, should be interpreted with caution. Prevalence of Salmonella infections determined in a single facility may not reflect the prevalence of Salmonella-infected horses in the general population; furthermore, obtaining a Salmonella isolate from a horse does not establish that the horse is a chronic Salmonella carrier. PMID- 10461636 TI - Lawsonia intracellularis-like organism infection in a miniature foal. AB - A 7-month-old foal was admitted to the hospital with a history of lethargy, weight loss, mild diarrhea, and anorexia. A diagnosis of proliferative enteritis caused by Lawsonia intracellularis-like organisms was made after necropsy and histologic examination of the small intestine. Although infection with L intracellularis-like organisms is a rare cause of enteritis in foals, it should be considered in the differential diagnosis, especially if the foal was housed in the proximity of pigs or pig feces. Antemortem diagnosis remains challenging because isolation of the organism in fecal material requires cell culture, and histologic evaluation of intestinal biopsy specimens may be unrewarding because of the lack of information regarding the frequency and distribution of lesions in horses. Alternatively, use of immunochemical stain, dot-blot technique, and polymerase chain reaction provide specific diagnostic tests that can be performed on fecal material. Postmortem diagnosis relies on histologic examination of infected tissues and use of immunofluorescence and polymerase chain reaction. PMID- 10461637 TI - Infertility and abortion among first-lactation dairy cows seropositive or seronegative for Leptospira interrogans serovar hardjo. AB - OBJECTIVE: To estimate the extent to which exposure to Leptospira hardjo before or at the time of first parturition was associated with infertility and abortion during the first lactation among dairy cows that had not been vaccinated for > or = 12 months. ANIMALS: 207 first-lactation cows from a herd of 2,000 lactating cows. PROCEDURE: Cows were tested for antibodies to L hardjo within 40 days after calving. Time from calving to first breeding, time from calving to conception, number of breedings per conception, and risk of abortion were compared between cows seropositive for L hardjo and cows that were seronegative. RESULTS: For the 9 (4.3%) cows that were seropositive for L hardjo, median time from calving to conception (132.6 days) was significantly longer than time for seronegative cows (95.4 days). Cows that were seropositive were twice as likely (relative risk, 2.07) to fail to conceive as seronegative cows. Mean number of breedings required per conception for seropositive cows (3.4) was significantly higher than that for seronegative cows (2.1). The proportion of seropositive cows that aborted was not significantly different from the proportion of seronegative cows that aborted. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure of nonvaccinated dairy cows to L hardjo can be associated with a subsequent reduction in fertility, as indicated by a greater time from calving to conception and higher number of breedings required per conception. The efficacy of leptospiral vaccines should be assessed to determine whether vaccination will minimize herd infertility associated with L hardjo infection. PMID- 10461638 TI - Congenital joint laxity and disproportionate dwarfism in a herd of beef cattle. AB - Three calves from a herd of beef cattle were examined because of disproportionate dwarfism and excessive extension of metacarpophalangeal and metatarsophalangeal joints. Abnormalities had been noticed at birth, 1 to 6 days earlier. A thorough herd investigation revealed that 16 calves born to 70 multiparous cows were affected during the calving season. The condition did not adversely affect calf survival. Affected calves had limbs that were disproportionally short, compared with their trunk size, and wide epiphyses of the femurs and humeri. Radiographic evaluation revealed incomplete maturation of carpal and tarsal bones and incomplete maturation and abnormal flaring of epiphyses of the short humeri and femurs. Histologic findings were consistent with chondrodystrophy. This disorder had not been seen in the herd in previous years and was traced to feeding of dry, spoiled silage to the dams during midgestation. Covering the silage prevented problems in the subsequent year. PMID- 10461639 TI - Management of androgenetic alopecia. AB - BACKGROUND: Androgenetic alopecia (AGA) is the most frequent cause of hair loss affecting up to 50% of men and 40% of women by the age of 50. METHODS: This paper outlines the current status of diagnosis and offers guidelines for optimal management of AGA in both men and women. RESULTS: The diagnosis of AGA can usually be confirmed by medical history and physical examination alone. A trichogram can be useful to assess the progression of the hair loss. A scalp biospy is diagnostic but usually not required. In women with signs of hyperandrogenism, investigation for ovarian (polycystic ovarian disease) or adrenal (late-onset congenital adrenal hyperplasia) disorders is required. Mild to moderate AGA in men can be treated with oral finasteride or topical minoxidil. Oral finasteride at the dosage of 1 mg/day produced clinical improvement in up to 66% of patients treated for 2 years. The drug is effective for both frontal and vertex hair thinning. Medical treatment with finasteride or minoxidil should be continued indefinitely since interruption of therapy leads to hair loss with return to pretreatment status. Mild to moderate AGA in women can be treated with oral antiandrogens (cyproterone acetate, spironolactone) and/or topical minoxidil with good results in many cases. Hair systems and surgery may be considered for selected cases of severe AGA both in men and in women. CONCLUSIONS: Patients with AGA should be informed about the pathogenesis of the condition. If used correctly, available medical treatments arrest progression of the disease and reverse miniaturization in most patients with mild to moderate AGA. PMID- 10461641 TI - Past and current understanding of the pathophysiology of muscle cramps: why treatment of varicose veins does not relieve leg cramps. AB - Historically relevant hypotheses on the pathophysiology of muscle cramps are reviewed. Psychosomatic, static, vascular, myogenic and neural theories are highlighted from a clinician's point of view. Modern neurophysiologic research leaves little doubt that muscle cramp is caused by excitation of spinal motor neurones mediated by changes in presynaptic input. Nevertheless, obsolete theories and relative treatments stubbornly persist in clinical practice. PMID- 10461640 TI - Notalgia paresthetica: clinical, physiopathological and therapeutic aspects. A study of 12 cases. AB - Notalgia paresthetica (NP) is a common but often unrecognized neurocutaneous condition, with very few cases reported to date. It is characterized by pruritus localized in an area between D2 and D6 dermatomes, sometimes accompanied by sensory neuropathies and/or electrical conductivity disorders. Cutaneous pigmented patches and friction amyloidosis can arise with irritation. Some hereditary cases have been noted mainly in young patients, associated with multiple endocrine neoplasia type 2A. However, NP mainly occurs in older patients and most are sporadic pathologies linked with musculoskeletal compression of spinal nerves. Only capsaicin has shown some (but unfortunately only transient) efficacy in relieving NP symptoms. We present observations on 12 sporadic cases of NP. Spinal X-rays revealed dorsal arthrosis or spinal static disequilibrium in nine of these patients. Six patients underwent spinal and paraspinal ultrasound or radiation (better) physiotherapy, and the symptoms subsided in four of these cases. These results highlighted that spinal disorders could be a determining factor in NP, indicating that patients could benefit from physiotherapy. PMID- 10461642 TI - Subcorneal pustular dermatosis involving the face. AB - We report an interesting case of subcorneal pustular dermatosis in a young woman with prominent involvement of the face. Facial involvement in this condition has not previously been reported. PMID- 10461643 TI - Multiple perifollicular fibromas. AB - Perifollicular fibroma is a benign mesodermal tumour of the hair follicle. It can occur as a solitary papule or as multiple lesions that are clinically indistinguishable from other tumours of the pilar apparatus. Multiple perifollicular fibromas may be inherited although the pattern remains unclear. Adnexal tumours can be associated with internal malignancy and perifollicular fibromas have been linked with adenomatous colonic polyps. This report describes a patient with multiple perifollicular fibromas with no associated malignancy to date and a family pedigree suggestive of an autosomal dominant pattern of inheritance. PMID- 10461644 TI - Aneurysmal fibrous histiocytoma: an unusual variant of cutaneous fibrous histiocytoma. AB - Aneurysmal fibrous histiocytoma (AFH) (Santa-Cruz DJ, Kyriakos M. Aneurysmal ('Angiomatoid') fibrous histiocytoma of the skin. Cancer 1981;47:2053-2061) is a distinct but poorly recognized clinicopathological variant of cutaneous fibrous histiocytoma (CFH) that may result from the slow extravasation of blood into the tumour. The resulting lesion can have a very different clinicopathological appearance resulting in diagnostic confusion. We describe a patient with an AFH that presented as a pigmented nodule on the foot and discuss clinical recognition and histological differentiation from other tumours. PMID- 10461645 TI - Herpetic whitlow in an AIDS patient. AB - This case confirms that cutaneous herpes simplex virus (HSV) infections in many AIDS patients is important not only for the difficulty in diagnosis of herpetic lesions, but also for the possibility that co-infection by HSV and HIV can adversely affect prognosis in these patients. PMID- 10461646 TI - Hydroxyurea-induced leg ulcers: is macroerythrocytosis a pathogenic factor? AB - Hydroxyurea is a common cancer chemotherapy agent that inhibits ribonucleotide reductase, an enzyme essential to DNA synthesis. It is considered the drug of choice in the treatment of chronic myelogenous leukemia and essential thrombocythemia. The occurrence of leg ulcers have been described in 8.5% of patients receiving continuous treatment with hydroxyurea, but the cause of this complication is unknown. We report two additional patients and suggest that macroerythrocytosis, which occurs in almost all the patients taking hydroxyurea, may be a pathogenic factor. Macroerythrocytosis can be considered as an 'acquired' blood dyscrasia, and similar leg ulcers have long been known to occur with certain hereditary blood dyscrasias, such as sickle cell anemia, thalasemia, and spherocytosis. PMID- 10461647 TI - Mid-dermal elastolysis associated with Hashimoto's thyroiditis. AB - We report the case of a 38-year-old Caucasian female presenting asymptomatic plaques of fine wrinkling and perifollicular papular protrusions especially on the trunk. Histological examination evidenced loss of elastic fibers in the mid dermis due to elastophagocytosis, with giant cells and granuloma formation. Moreover, elevated titers of thyroid autoantibodies were detected and thyroid ultrasound revealed echo-poor tissue. These findings met the diagnoses of mid dermal elastolysis and Hashimoto's thyroiditis. This association has not been reported before. We present a comprehensive overview of the literature and discuss the pathogenetic aspects of mid-dermal elastolysis and the significance of the association with Hashimoto's thyroiditis. PMID- 10461648 TI - Asthma induced by allergy to Trichophyton rubrum. AB - The worldwide incidence of asthma and of allergic respiratory diseases is increasing (Akiyama K. 'Environmental allergens and allergic diseases.' Rinsho Byori 1997;45(1):13. D'Amato G, Liccardi G, D'Amato M. Environment and development of respiratory allergy. II. Indoors. Monaldi Arch Chest Dis 1994;49(5):412. Weeke AR. Epidemiology of allergic diseases in children. Rhinol Suppl 1992;13:5. Ulrik CS, Backer V, Hesse B, Dirksen A. Risk factors for development of asthma in children and adolescents: findings from a longitudinal population study. Respir Med 1996;90(10):623.) This has been attributed to several factors, including lifestyle changes and an expanding variety of potential causative allergens. Management of asthma entails preventive and acute medications, immunologic therapies, and removal of the identified allergen(s) from the patient's environment. Without the latter, patients may not experience full symptomatic relief. This case report describes a patient who developed bronchial asthma subsequent to an infection of tinea pedis and pedal onychomycosis; antifungal management resulted in full resolution of his tinea pedis, onychomycosis and asthma. PMID- 10461649 TI - Subcutaneous fat necrosis of the newborn. AB - Subcutaneous fat necrosis of the newborn (SFN) is an uncommon disease that affects newborns who have suffered from tissue hypoxia during or following delivery. This disease appears during the first weeks of life. It consists of indurate, erythematous or purple-erythematous nodules and plaques in the skin. Histology of a biopsy specimen shows granulomatous necrosis in the subcutis with radial crystals in lipocytes and giant cells. Spontaneous resolution in a few weeks is usual, but the mobilization of calcium from the necrosed subcutis together with the action of some hormones may cause hypercalcemia and certain serious complications. A newborn female child developed SFN after dystocic delivery causing cerebral frontal lobe hemorrhage. The skin nodules resolved spontaneously in a few weeks and no complications were observed 1 year later. PMID- 10461650 TI - Primary cutaneous mucormycosis: a diagnosis to consider. AB - Primary cutaneous mucormycosis is a deep fungal infection, mainly seen in diabetics and immunocompromised subjects. Rapid diagnosis and therapy are necessary to avoid fatal outcome. We describe the complete histopathological and microbiological studies of primary cutaneous mucormycosis in a 74-year-old man with several risk factors, such as chronic obstructive pulmonary disease, respiratory acidosis, hemolytic anemia, myelodysplastic syndrome and iatrogenic diabetes, due to corticosteroid therapy. He developed two cutaneous necrotic scars on his left leg. Mucormycosis was suspected and specimens from surgical debridement were histopathologically and microbiologically studied confirming the clinical diagnosis. Amphotericin B was given topically and intravenously resulting in complete healing of the ulcer. Risk factors and microbiological studies are compared with those in the current literature. It is necessary in certain cases to suspect mucormycosis infections in diabetics, immunocompromised subjects and even in healthy individuals. Rapid diagnosis and treatment are important, but they should be based on complete histopathological and microbiological studies, to establish the genus of the causal agent. PMID- 10461651 TI - The inter-relationship between isotretinoin/acne and depression. PMID- 10461652 TI - Soluble ICAM-1 seems to be related to prognosis in malignant melanoma. PMID- 10461653 TI - Uncommon presentation of cutaneous leishmaniasis as eczema-like eruption. PMID- 10461654 TI - Primary cutaneous Aspergillosis in a patient with acquired immunodeficiency syndrome. PMID- 10461655 TI - Annual recurrent annular acroerythema without lactate dehydrogenase M-subunit deficiency. PMID- 10461656 TI - Erysipelas melanomatosum. PMID- 10461657 TI - Borderline tuberculoid leprosy coexisting with systemic lupus erythematosus. PMID- 10461658 TI - Malignant pyoderma. PMID- 10461659 TI - Xanthoma disseminatum with multi-system involvement and fatal outcome. PMID- 10461660 TI - Periorbicular discoid lupus erythematosus. PMID- 10461661 TI - Scar sarcoidosis following herpes zoster. PMID- 10461662 TI - Generalized morphea and lichen sclerosus et atrophicus successfully treated with sulphasalazine. PMID- 10461663 TI - Positivism and psychoanalysis: a new look at the historical record. PMID- 10461664 TI - Variations on a theme of shame: Chekhov, Glenn Gould, and the "cased-in-man" syndrome. PMID- 10461665 TI - Psychoanalysis as askesis. PMID- 10461666 TI - Inauthentic communication and the false self. PMID- 10461667 TI - Freud, Moses, and the religions of Egyptian antiquity: a journey through history. PMID- 10461668 TI - Numbers and psychoanalysis: reflections on the quest for certainty. 2. PMID- 10461669 TI - The riddle that doesn't exist: Ludwig Wittgenstein's transmogrification of death. PMID- 10461670 TI - Laboratory production of buffalo (Bubalus bubalis) embryos. AB - There is an increasing interest in large-scale in vitro production (IVP) of buffalo embryos through in vitro maturation (IVM), fertilization (IVF) and culture (IVC) of oocytes for faster multiplication of superior germplasm. The recovery of total and usable quality oocytes from slaughterhouse ovaries is low in this species. The nuclear maturation rates of buffalo oocytes matured in the presence of follicular fluid or serum and hormones like luteinizing hormone, follicle-stimulating hormone and oestradiol vary from 70 to 80% and are comparable to those reported for cattle oocytes. However, with fertilization rates of 40-55%, and the yield of blastocysts at around 10-15%, the efficiency of IVP is much lower than that in cattle. The in vitro sperm preparation procedures and the systems employed for performing IVF and culture of zygotes up to blastocyst stage are suboptimal and need substantial improvements. The quality and viability of blastocysts produced need to be checked by cell count, and after transfer to synchronized recipients, for development of quality control standards. PMID- 10461671 TI - Developmental changes in plasma angiotensin-converting enzyme concentration in fetal and neonatal lambs. AB - Relationships between plasma angiotensin-converting enzyme (ACE) and cortisol, and blood pressure were examined in chronically catheterized ewes and their fetuses during late gestation (111-141 days, term 145 +/- 2 days). Plasma ACE was also measured in non-pregnant adult ewes and in lambs over the first 5 weeks of life. In fetuses near term (136-141 days), plasma ACE was greater than in those studied earlier in gestation; overall, plasma ACE correlated with gestational age (r = 0.72). The ontogenic rise in plasma ACE was associated with prepartum increases in plasma cortisol (r = 0.67) and blood pressure (r = 0.66). No relationship was observed between plasma ACE and partial pressure of oxygen in utero. Peak plasma ACE concentration observed in fetuses near term was maintained in newborn lambs for 3 days after birth. By 2 weeks of postnatal age, plasma ACE had decreased to the value seen in non-pregnant adult ewes. Maternal plasma ACE was similar at all gestational ages studied, and was lower than that observed in non-pregnant ewes. Therefore, in the sheep fetus, plasma ACE increased towards term in association with the prepartum cortisol surge. Developmental changes in ACE activity may be partly responsible for the ontogenic rise in fetal blood pressure. PMID- 10461672 TI - Gestational changes in fetal renal and hepatic angiotensinogen mRNA and protein. AB - The aim of the study was to determine the amount of angiotensinogen expression and its protein product in fetal sheep liver and kidney in the last third of gestation. Angiotensinogen mRNA was measured by RNase protection assay and its protein levels were measured by radioimmunoassay. Levels were measured at 80, 95, 111, 125 and 139 days. Angiotensinogen mRNA was present in all fetal liver and kidney samples tested. The ratio of hepatic angiotensinogen mRNA/18 S rRNA increased by 100% (P < 0.001) and angiotensinogen levels increased by 33% (P < 0.001) in fetal sheep from 80 to 139 d. Over the same period the ratio of renal angiotensinogen mRNA/18 S rRNA increased by 170% (P < 0.001) and renal angiotensinogen protein increased by 41% (P < 0.001). The levels of angiotensinogen mRNA and its protein in the adult kidney were less than in kidneys of 139 d old fetuses (P < 0.01). There was a direct relationship between levels of angiotensinogen mRNA and its protein in the liver (r = 0.53, P < 0.01, n = 25) and in the kidney (r = 0.75, P < 0.0001, n = 24). These findings demonstrate that there is a significant increase in both hepatic and renal angiotensinogen gene expression in the last third of gestation in the fetal sheep and that this increase is associated with an increase of angiotensinogen levels in both tissues. This increase in angiotensinogen in late gestation could influence the activity of both the intrarenal and circulating renin angiotensin systems. PMID- 10461673 TI - Low-dose flunarizine does not affect short-term fetal circulatory responses to acute asphyxia in sheep near term. AB - Asphyxia is one of the major causes of perinatal brain damage and neuronal cell loss, which may result in psychomotor deficits during later development. It has been shown previously that the immature brain can be protected from ischemic injury by flunarizine, a class IV calcium antagonist. However, cardiovascular side-effects of flunarizine, when applied at the dosages used in those studies, have been reported. Recently, the present authors were able to demonstrate that even by injecting flunarizine at a far lower dosage (1 mg kg-1 estimated bodyweight) neuronal cell damage, caused by occlusion of both carotid arteries for 30 min, can be reduced in fetal sheep near term. The aim of the present study was, therefore, to examine whether low-dose flunarizine affects fetal cardiovascular responses to acute asphyxia in sheep near term. Ten fetal sheep were chronically instrumented at a mean gestational age of 132 +/- 1 days (term is at 147 days). Fetuses from the study group received a bolus injection of flunarizine (1 mg kg-1 estimated fetal weight) 60 min before asphyxia, whereas the solvent was administered to the fetuses from the control group. Organ blood flows, physiological variables and plasma concentrations of catecholamines were measured before, during and after a single occlusion of uterine blood flow for 2 min (i.e. at 0, 1, 2, 3, 4, and 30 min). Before asphyxia, the distribution of combined ventricular output and physiological variables, as well as concentrations of catecholamines, in fetuses from the control group were in the normal range for chronically prepared fetal sheep near term. During acute asphyxia there was a redistribution of cardiac output towards the central organs accompanied by a pronounced bradycardia and a rapid increase in arterial blood pressure. After asphyxia circulatory centralization did not resolve quite as rapidly as it developed, but was almost completely recovered at 30 min after the insult. There were nearly no differences in the time course of physiological and cardiovascular variables measured before, during and after acute intrauterine asphyxia between the control and study groups. From the present study it was concluded that low-dose flunarizine does not affect short-term fetal circulatory responses to acute asphyxia in sheep near term. PMID- 10461674 TI - Successful development in vitro of hamster 8-cell embryos to 'zona-escaped' and attached blastocysts: assessment of quality and trophoblast outgrowth. AB - The peri-implantation development involves zona escape (hatching) of blastocysts and their attachment and proliferation. These events are difficult to study in vivo, so in this study hamster 8-cell embryos were cultured through the hatched and attached blastocyst stages using different formulations of hamster embryo culture medium (HECM)-2. Supplementation of succinate, amino acids, vitamins (inositol, pantothenate, choline chloride) and bovine serum albumin (BSA) to HECM 2 supported 100% development of 'zona-escaped' blastocysts. In this medium (designated as hatching, i.e. HECM-2h) all blastocysts invariably deflated and escaped from focally lysed zonae, which underwent complete dissolution. In their presence, pre-morula stage embryos also escaped from zonae. Omission of BSA from HECM-2h failed to support zona escape whereas that of vitamins reduced zona escape (34.0% +/- 7.0). Blastocysts with the potential to undergo zona escape in HECM-2h were of high quality as they had a higher mean cell number (MCN) than the MCN of those developing in BSA-free HECM-2h (35.2 +/- 1.6 v. 24.3 +/- 1.1). Cell allocation (i.e. trophectoderm to inner cell mass ratio) in blastocysts remained unaltered in both media (2.6 +/- 0.2 v. 2.7 +/- 0.2). Supplementation of 10% bovine fetal serum (BFS) to HECM-2h was detrimental to the development of blastocysts (22.3% +/- 7.4) and none of them underwent zona escape. Interestingly, BFS was required either as a supplement to the medium or as a coating on dishes for azonal blastocysts to attach (> or = 70%) and exhibit trophoblast (TB) outgrowth (30.3 x 10(3) +/- 2.9 x 10(3) micron 2 at 48 h). These results show that HECM-2h supports maximal development of zona-escaped blastocysts with the potential to attach and exhibit TB outgrowth, and there is a developmental stage-specific requirement for serum during peri-attachment in hamster development. PMID- 10461675 TI - Increased ovulation rate in Merino ewes immunized against small synthetic peptide fragments of the inhibin alpha subunit. AB - Four experiments were carried out in Merino ewes during a period of 4 years to determine the long-term effects of immunization against different synthetic peptides mimicking the amine terminal of the alpha subunit of porcine inhibin. Peptides were conjugated to human serum albumin and 100-200 micrograms emulsified in Freund's complete adjuvant for the primary immunization. Usually two booster injections were given at monthly intervals with 50-100 micrograms conjugated peptide using either incomplete Freund's adjuvant or Montanide:Marcol. In some experiments a further immunization was carried in the next year. Blood samples were taken 10 days after each immunization, during the luteal phase, for estimation of gonadotrophin concentrations and determination of inhibin antibody titres. One day after blood sampling cloprostenol was used to induce luteolysis and laparoscopy was performed in the subsequent oestrous cycle. Immunization of ewes with synthetic peptides 1-32, 1-26, 7-26 and 8-30 resulted in large increases in the ovulation rate (OR). An approximately two-fold increase in OR was observed following the first booster immunization with these peptides and a three- to five-fold increase after the second booster immunization. Immunization with these large peptides resulted in a sustained increase in OR for a period of at least 1 year after the second booster immunization. Of the shorter peptides, peptides 10-26 and 13-26 gave a reasonable ovulatory response, although it was more difficult to obtain a response with peptides 1-16, 8-22, 13-25, 8-19 and 10 19; peptides 7-13 and 1-6 gave no response (but were examined for one breeding season only). The smaller peptides led to lower inhibin antibody titres that were not necessarily associated with increased follicle-stimulating hormone (FSH) or OR. More intensive blood sampling in one experiment showed that following primary immunization against peptide 1-32 there was a transient increase in plasma FSH, which did not lead to an increased OR. Moreover, a prolonged period of raised FSH after the first booster was significantly correlated with increased OR. In these animals antibody titres were only slightly increased after primary immunization, but after the first booster immunization higher titres were observed that were significantly correlated with trough FSH values and the subsequent OR. These results are interpreted as showing that (1) to obtain an increase in OR peptides 1-32, 1-26 and 7-26 are suitable as immunogens; (2) smaller peptides are less reliable, often require multiple injections, and the response may be delayed; and (3) an extended period of raised plasma FSH is needed to give a large ovulatory response. PMID- 10461676 TI - The relationship between membrane status and fertility of boar spermatozoa after flow cytometric sorting in the presence or absence of seminal plasma. AB - The motility, viability (percent live), capacitation status and in vitro fertility of boar spermatozoa were examined, after staining with Hoechst 33,342 and flow cytometric sorting in the absence or presence of seminal plasma. Viability was higher in unstained controls and when seminal plasma was present in the medium used to collect spermatozoa from the cell sorter than when seminal plasma was absent or in the staining extender only, but motility was highest when seminal plasma was included in the extender only, compared with the controls and other treatments. The proportions of capacitated spermatozoa were increased by sorting, but were lower when seminal plasma was present, rather than absent, from the staining extender and the collection medium. Compared with unstained controls, extension and staining without sorting only increased the proportion of capacitated spermatozoa after washing in preparation for in vitro fertilization. The percentages of polyspermic, penetrated and cleaved oocytes were lower when inseminated with unsorted (stained) than control (unstained) spermatozoa, regardless of the presence or absence of seminal plasma. These parameters were higher for sorted than for control spermatozoa in the absence of seminal plasma, but in its presence penetration and cleavage were substantially lower. The proportions of capacitated spermatozoa were lower when seminal plasma was present in the collection medium only than in the staining extender or when it was absent altogether, but the former treatment substantially reduced the proportions of polyspermic, penetrated and cleaved oocytes, and the proportion of blastocysts. These findings indicate that sperm capacitation associated with flow cytometric sorting can be reduced by the inclusion of seminal plasma in the collection medium, but this treatment reduces the ability of spermatozoa to fertilize oocytes in vitro under these conditions. PMID- 10461677 TI - Comparison of the merits of measuring equine chorionic gonadotrophin (eCG) and blood and faecal concentrations of oestrone sulphate for determining the pregnancy status of miniature horses. AB - The relative merits of measuring blood concentrations of equine chorionic gonadotrophin (eCG, previously known as pregnant mare serum gonadotrophin (PMSG)), or oestrone sulphate (OS), or faecal OS concentrations for determining pregnancy status in miniature horses were investigated. Pregnant mares between 40 and 140 days after mating had serum eCG concentrations > 1 I.U. mL-1, with the highest concentrations occurring between days 50 and 120. However, eCG measurements were susceptible to returning a 'false positive' diagnosis of pregnancy. Plasma OS concentrations ranged from 0.1 to 3.6 ng mL-1 in non pregnant mares, whereas pregnant mares beyond 100 days post-mating all had plasma OS concentrations > 30 ng mL-1. Faecal OS concentrations ranged from 4 to 89 ng g 1 in non-pregnant mares. For faecal samples collected from pregnant mares 150 days or more after mating, 97% of samples had OS concentrations > 85 ng g-1, the value 3 standard deviations above the mean non-pregnant value. None had values below 67 ng g-1, the value 2 standard deviations above the mean non-pregnant value. These results show that measurement of eCG is suitable for determining pregnancy status in miniature mares between 40 and 100 days post-mating. However, mares returning a 'pregnant' diagnosis should undergo a blood OS test 100 or more days after mating to eliminate the possibility of a 'false positive' diagnosis. Measuring blood OS is recommended as the method of choice for determining pregnancy status in miniature mares 100 or more days after mating. Faecal OS measurements provide a non-invasive alternative to blood OS testing from 150 days post-mating. However, the discrimination between 'pregnant' and 'non-pregnant' levels of OS is not as great in faeces as it is in blood. PMID- 10461679 TI - Oxytocin does not induce a rise in intracellular free calcium in human breast cancer cells. AB - Research suggests that oxytocin acts as a growth modulating agent for breast cancer cells. However, the signaling mechanisms responsible for these modulatory effects have not been fully elucidated. In the physiological setting oxytocin is known to stimulate contraction of myometrial cells in the uterus and myoepithelial cells in the breast by increasing intracellular free calcium ([Ca2+]i). The expression of oxytocin receptor mRNA in T-47D breast cancer cells, and four additional breast cancer cell lines (BT-549, MCF-7, MDA-MB- 231, ZR-75 1), was confirmed by RT-PCR analysis. Oxytocin-induced changes in [Ca2+]i in indo 1 AM loaded T-47D breast cancer cells were monitored using flow cytometric analysis. In this cell line, oxytocin (0, 1, 10, 100, and 1,000 nM) did not induce a dose-dependent increase in the mean 405 nm/485 nm emission ratio. These results indicate that oxytocin signaling in T-47D breast cancer cells does not appear to involve an increase in [Ca2+]i. PMID- 10461678 TI - Effects of calcium and bicarbonate on head-to-head agglutination in ejaculated boar spermatozoa. AB - The present study was conducted to reveal the effects of calcium and bicarbonate on the occurrence of head-to-head agglutination in ejaculated boar spermatozoa in vitro. Boar spermatozoa were washed and incubated in a modified Krebs-Ringer bicarbonate (mKRB) in a 37 degrees C CO2 incubator (5% CO2 in air) for 1-5 h. Before and after the incubation, aliquots of each sperm sample were fixed, smeared on glass slides, and stained with a phosphate-buffered solution of Giemsa to assess the percentages of head-to-head agglutinated spermatozoa. Before the incubation, only 5-12% of the spermatozoa were agglutinated. After the 1-h incubation, however, the percentage of head-to-head agglutinated spermatozoa rose to approximately 50%, followed by only minor increases thereafter. This rise was dependent on the concentrations of calcium chloride contained in the mKRB and was attenuated by the addition of 2 mM [ethylenebis(oxyethylenenitrilo)]tetra-acetic acid (EGTA) to the medium. Moreover, the replacement of sodium bicarbonate with 2 [4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid (Hepes) in the medium and treatment with ruthenium red, which have both been shown previously to inhibit calcium uptake by boar spermatozoa, significantly reduced the rise. Based on these findings, it was concluded that extracellular calcium and bicarbonate are key factors regulating head-to-head agglutination in boar spermatozoa. The possible relationship between agglutinability and the fertilizing ability of boar spermatozoa is also discussed. PMID- 10461680 TI - Effects of selective and non-selective endothelin antagonists on ischemia reperfusion damage in the isolated perfused murine liver. AB - The objective of the present study was to clarify the differential effects of endothelinA (ETA) and ETB antagonism in the early phase of ischemia-reperfusion damage. Male Sprague Dawley rats were randomly divided into 4 groups: control (n = 10), bosentan (40 nM; n = 10), BQ-485 (20 nM; n = 10), and BQ-788 (50 nM; n = 10) to compare the effects of ETA or ETB or both ETA and ETB antagonists against the warm ischemia-reperfusion damage of murine livers. Isolated livers were perfused with oxygenated Krebs-Henseleit bicarbonate buffer solution and ET antagonists for 30 min before inducing warm ischemia (non-recirculating system). After 40 min without perfusate, measurements (portal pressure, O2 tensions of influent and effluent perfusate, liver enzymes, etc.) were taken up to 60 min after reperfusion. The BQ-788 group had significantly more liver damage than did the other groups, and more O2 consumption than did the bosentan group. BQ-485 and bosentan were more protective at some points after reperfusion. Antagonism of only the ETB receptors is detrimental, but antagonism of the ETA receptors appears to have a role in protecting the liver from warm ischemia-reperfusion damage in the early phase. PMID- 10461681 TI - Neutrophil elastase enhances macrophage production of chemokines in receptor mediated reaction. AB - Proteases influence various leukocyte effector functions. We investigated the specific binding activity of neutrophil elastase (NE) on rat peritoneal macrophages to stimulate chemokine production in vitro. NE enhanced macrophage production of monocyte chemo-attractant protein-1 (MCP-1) (CC-chemokine) and cytokine-induced neutrophil chemo-attractant (CINC) (CXC-chemokine). However, the serine protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), significantly reduced these chemokine productions by macrophages stimulated with NE. 125I-NE was significantly bound to macrophages, but 125I-NE binding was inhibited by addition of unlabeled NE. In addition, NE significantly stimulated [3H]thymidine incorporation into DNA on 3T3 fibroblasts in a dose-dependent manner. These results suggest that NE stimulates chemokine production by macrophages. This may be a receptor-mediated reaction. Thus, NE not only degrades extra-cellular matrix but also stimulates chemokine production by macrophages. PMID- 10461682 TI - Micromethods for the analysis of tear proteins in pharmacological studies. AB - Although it is well established that ocular mucins and other proteins are essential for tear film stability, whether certain drugs, like non steroidal antiinflammatory drugs (NSAIDs), could cause ocular dryness by inhibiting their secretion is not known. To perform these and other studies of pharmacological interest, we evaluated several micromethods for the analysis of tear samples. The major proteins of the tear fluid collected in capillaries, i.e. IgA, lactoferrin, tear specific prealbumin and lysozyme, were analyzed by SDS-PAGE and gel permeation HPLC, using 2.5-5 microliters of sample. Gastric mucin (PGM), examined as a standard, was analyzed by solid phase assays based on previously described histochemical staining methods: dot blot assays were performed using small disks of polyvinylidene difluoride or nylon membranes, prepared by an ordinary paper punch, which were coated with PGM and stained by Alcian blue or the periodic acid Schiff's reagent. The densitometric analysis was carried out using an ordinary flat scanner controlled by a personal computer equipped with an inexpensive software. The sensitivity of these simple assays was low (100-500 micrograms) but considered sufficient for certain studies. A more sensitive assay (5-20 micrograms) was developed by immobilizing PGM in small agarose gels (100 microliters), prepared in the wells of 96-well microplates, which could by stained by stains-all and analyzed by an automatic plate reader at 595 nm. PMID- 10461683 TI - Preventive effect of zinc compounds, polaprezinc and zinc acetate against the onset of hepatitis in Long-Evans Cinnamon rat. AB - It is known that Long-Evans Cinnamon (LEC) rats are characterized by the fulminant hepatitis occurring as a result of an abnormal hepatic deposition of Cu due to the lack of the Cu-transporter p-type ATPase. To prevent the hepatitis, two Zn compounds, Zn acetate and polaprezinc were given orally to LEC rats aged 30 days. At 100 days after birth, the control group composed of LEC rats fed a basal diet (Cu, 17 ppm; Zn, 50 ppm; Fe, 150 ppm) exhibited slight jaundice and showed high activities of serum enzymes related to hepatic function. The groups fed the diet fortified (1000 ppm as Zn) with Zn acetate or polaprezinc did not have jaundice. The hepatic Cu concentrations were 174 +/- 34 micrograms/g and 156 +/- 23 micrograms/g in the polaplezinc group and Zn acetate group, respectively. The control group showed 267 +/- 17 micrograms Cu/g and 298 +/- 62 micrograms Fe/g in the liver. The Fe concentration was about 1.7 times the concentration in the two Zn groups. Hepatic free Cu and Fe concentrations were 2.6 +/- 0.3 and 21.4 +/- 5.8 micrograms/g, 1.7 +/- 0.7 and 6.8 +/- 1.1 micrograms/g, and 1.3 +/- 0.1 and 6.2 +/- 0.8 micrograms/g in the control, polaprezinc and zinc acetate groups, respectively. Intestinal metallothionein (MT) concentrations were not increased significantly by the Zn diets. The two Zn compounds inhibit Cu absorption from the intestinal tract, resulting in a decrease of hepatic Cu deposition. The new Zn compound as well as Zn acetate is categorized as a therapeutic drug for Cu poisoning, including Wilson's disease. PMID- 10461684 TI - Therapeutic effects of tetrathiomolybdate on hepatic dysfunction occurring naturally in Long-Evans Cinnamon (LEC) rats: a bona fide animal model for Wilson's disease. AB - Long-Evans Cinnamon (LEC) rats were fed a diet containing 7 ppm Cu and 30 ppm Cu from 60 days after birth. Fischer (Fischer group) and LEC (LEC-control group) rats fed a 7 ppm Cu diet showed normal growth throughout the whole period (60 to 125 days after birth). On the other hand, LEC rats fed the 30 ppm Cu diet had decreased body weight and showed slight jaundice at around 100 days after birth. Tetrathiomolybdate (TTM, 10 mg/kg bw) was injected sub-cutaneously at 101 and 105 days after birth into half of the LEC rats fed the 30 ppm Cu diet. LEC rats given TTM (LEC+TTM group) recovered their body weight and the jaundice rapidly disappeared. However, LEC rats without TTM (LEC-TTM group) had sharply decreased body weight and showed severe jaundice at 103 days after birth. The hepatic Cu concentration in LEC+TTM rats (460 micrograms/g) exceeded that of LEC-control rats (330 micrograms/g) at 125 days after birth. Our data suggest that TTM is effective for treatment of acute hepatic injury in the LEC rat. PMID- 10461685 TI - Copper increases in both plasma and red blood cells at the onset of acute hepatitis in LEC rats. AB - Ceruloplasmin is excreted mostly in the apo-form in Wilson's disease patients and Long-Evans rats with a cinnamon-like coat color (LEC rats), an animal model for Wilson's disease, and hence the concentration of Cu in the plasma is low. However, it increases toward and at the onset of acute hepatitis in LEC rats, the increased Cu in the plasma being bound to ceruloplasmin, metallothionein and albumin. Changes in the concentration of Cu in red blood cells (RBCs) were monitored with age for the first time together with that in the plasma in LEC rats. Cu in the RBCs was found to increase to a 5-7 times higher level than that in the plasma toward the onset and peaked at the onset, the pattern being similar to that in the plasma. The source of the Cu increase in the RBCs was discussed, and it was assumed that the so-called free Cu ions that leak from the damaged hepatocytes are bound to albumin and/or taken up by the RBCs. PMID- 10461686 TI - Insulin resistance and impaired endothelium-dependent renal vasodilatation in fructose-fed hypertensive rats. AB - We investigated the contribution made by hyperinsulinemia and endothelial dysfunction to the hypertension of rats that had received 10% fructose in their drinking water for 12 weeks. As control animals with endothelial dysfunction, we used streptozotocin-induced diabetic rats. Measurements were made at 12 weeks after the start of fructose feeding or a single streptozotocin injection. Systolic blood pressure was greater in fructose-fed rats than in either streptozotocin-diabetic rats or age-matched controls. By comparison with the age matched controls, the plasma levels of glucose, triglyceride, high density lipoprotein cholesterol, low density lipoprotein cholesterol and free fatty acids were all significantly raised in both fructose-fed rats and streptozotocin diabetic rats. The plasma insulin was significantly raised in fructose-fed rats, whereas it was significantly lowered in streptozotocin-diabetic rats. The vasodilatation induced in the perfused kidney by acetylcholine was weaker in both fructose-fed rats and streptozotocin-diabetic rats than in the age-matched controls and these weakened responses were further attenuated by indomethacin. Acetylcholine increased the nitrite and nitrate (NO2- and NO3-) levels in the renal perfusate in age-matched controls. This effect was much weaker in the two experimental groups. These results suggest that endothelial dysfunction in fructose-fed rats and streptozotocin-diabetic rats may be due to a decreased synthesis of nitric oxide at least in the perfused kidney. It is further suggested that hyperinsulinemia is more important than endothelial dysfunction as a cause of hypertension in fructose-fed rats. PMID- 10461687 TI - Pharmacokinetics of gliquidone, glibenclamide, gliclazide and glipizide in middle aged and aged subjects. AB - Six middle-aged (42-59 years old) and six aged (71-75 years old) subjects received each, on separate days, an oral administration of gliquidone (30 mg), glibenclamide (5 mg), gliclazide (80 mg) and glipizide (5 mg). The plasma concentration of the drugs was measured before and at eight times (60 min to 24 h) thereafter. The half-life of gliclazide was higher than that of the other three hypoglycemic agents in middle-aged subjects and was the sole to be significantly increased in aged subjects. There is no obvious difference between sulfonylureas eliminated mainly by either the kidney (glibenclamide, gliclazide, glipizide) or the liver (gliquidone) in terms of the influence of aging upon their clearance. PMID- 10461688 TI - Screening for colorectal cancer. PMID- 10461689 TI - Crohn's disease in childhood: the case for enteral nutrition. PMID- 10461690 TI - Maximising the quality of care in rheumatoid arthritis. PMID- 10461691 TI - Scotland's health service and the voluntary sector. PMID- 10461692 TI - Royal College of Physicians of Edinburgh Consensus Conference on lipid lowering to prevent vascular events 17 & 18 March 1999. PMID- 10461693 TI - The Football World Cup 1998: an analysis of related attendances to an accident and emergency department. AB - Throughout the Football World Cup Finals of France 1998, patients attended our inner city Accident and Emergency department with a variety of conditions related to the event. Analysis was made on the total of 151 patients who attended with World Cup related presentations during this five-week period. The majority were young men involved in alcohol related trauma. The workload of the department was only substantially affected after one match, that of the opening game between Brazil and Scotland. PMID- 10461694 TI - Polaroid photographic referral for skin cancer--a potentially useful method of reducing time to surgery. AB - A prospective trial was conducted on twenty-five patients referred by dermatologists during 1998 using a referral letter and an accompanying polaroid photograph for prospective plastic surgical management of skin cancers. Using the description and the photograph, suitable patients were given dates for operation without requiring a plastic surgery out-patient clinic appointment. This enabled, not only a saving in the cost of an out-patient appointment, but a significant reduction in waiting time to surgery. PMID- 10461695 TI - Immediate psychiatric discharge letters by fax. AB - Much psychiatric care is provided outside the hospital setting. It is important for general practitioners (GPs) to have available information of good quality, provided promptly, after patients' discharges from in-patient psychiatric care to enable them to provide a high standard of follow-up care. In order to assess the value of hand-written Immediate Discharge Letters sent by fax we undertook a postal questionnaire survey of GPs, and examined a proportion of the clinical notes relating to 160 patients who between January and March 1998 had been discharged from in-patient care in the psychiatry admission wards at Crichton Royal Hospital, Dumfries. We found that structured letters, hand-written on a patient's discharge from in-patient status were generally valued by GPs as was their transmission by fax. Though certain deficiencies were confirmed in their completion, they are of value pending the arrival of a more definitive final discharge summary. We conclude that the continued use of such immediate discharge letters in psychiatry and their continued transmission by fax is justified. PMID- 10461696 TI - Primary ciliary dyskinesia syndrome and primary generalised epilepsy. AB - A child with primary ciliary dyskinesia syndrome (PCDS) developed primary generalised epilepsy (PGE). The PGE had characteristics of both childhood absence epilepsy (CAE) and juvenile myoclonic epilepsy (JME). The association of PCDS and PGE has not previously been recorded. PMID- 10461697 TI - An audit of paediatric audit in Scotland. AB - We undertook a retrospective review of the Scottish Clinical Audit Project Register (SCAPR). This generated information on audit activities in Scotland relating to children aged 0-15 years over the period 1992 to 1997. All healthcare specialties (including dentistry) in both acute and community sectors were included. Of the 7,073 registered entries, 613 (9%) relating to children were identified. Greater Glasgow contributed a third of entries despite having only 18% of the child population. Over half the projects were conducted in secondary care settings and only 5% in primary care. Medicine and nursing appeared the most audit active professional groups. Our findings suggest that children (who comprise 19% of the Scottish population) are an under-audited group in Scotland. Secondary care (especially surgery) seemed over-represented and primary care under-represented as audit settings. We were unable to use the SCAPR to identify specific service deficiencies or improvements that may have been stimulated in the course of "closing the audit loop." Future refinements to the way data are collected and recorded on the SCAPR will clearly require to address this shortcoming. PMID- 10461698 TI - Isaac Israeli and his Book of Urine. AB - Isaac Israeli (c850-950), philosopher and physician was born in Egypt where he practised as an oculist before moving to Kairouan, in Tunisia, where he served as physician to the Caliph Ziyadat Allah and later to 'Ubayd Allah al-Mahdi. His best known medical work Book of Fevers (Kitab al-Hummayat) was widely used as a medical text until the seventeenth century. This paper reviews Israeli's medical career through his Book of Urine (Kitab al-bawl), based on a fifteenth century Hebrew manuscript Sefer Hasheten (MS Hunter 477) in the Hunterian Collection of the University of Glasgow. This manuscript, most likely a translation from the original Arabic, describes the formation of urine and the value of visual examination of urine for the diagnosis and prognosis of disease. Israeli emerges as an outstanding physician and scholar who made a lasting contribution to Arab Jewish medicine in its most productive period. PMID- 10461699 TI - Metastatic epidural spinal cord compression. AB - OBJECTIVES: To provide an overview on the presentation, diagnosis, and treatment of metastatic epidural spinal cord compression (MESCC). DATA SOURCES: Published articles, book chapters, and research reports. CONCLUSIONS: MESCC is a common oncologic emergency that requires prompt recognition and emergency treatment to relieve pain and preserve neurologic function. The signs and symptoms of MESCC are easily detected and can be integral assessment components of the nursing care of any patient with a solid tumor. IMPLICATIONS FOR NURSING PRACTICE: Nurses can have a dramatic impact on preventing neurologic complications caused by this oncologic metastatic problem. When neurologic compromise is not prevented or reversed, nurses also can provide expert care to patients and families in the rehabilitation phase of MESCC. PMID- 10461700 TI - Syndrome of inappropriate secretion of antidiuretic hormone in malignancy. AB - OBJECTIVES: To provide a review of pathophysiology, epidemiology, signs and symptoms, diagnosis, treatment, and nursing management of the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) in malignancy. DATA SOURCES: Research reports, review articles, and book chapters relevant to SIADH. CONCLUSIONS: The principle cause of hyponatremia in malignancy is SIADH. Early recognition and prompt treatment can prevent serious neurologic sequelae. Antineoplastic therapy and methods to correct hyponatremia constitute effective treatment strategies. IMPLICATIONS FOR NURSING PRACTICE: Nurses play a major role in nursing assessment, appropriate interventions and symptom control, and promoting resolution of problems and optimal quality of life in patients with malignancy complicated by SIADH. PMID- 10461701 TI - Cardiac tamponade. AB - OBJECTIVES: To review the oncologic emergency of cardiac tamponade through a case study presentation/analysis and a discussion of the pathophysiology, diagnosis, treatment, and nursing management. DATA SOURCES: Research studies, review articles, book chapters, abstracts, and clinical practice. CONCLUSIONS: Cardiac tamponade is a potentially life-threatening condition that is not uncommon in the oncology setting. It can result directly from the malignant or metastatic process or from the treatment of the malignancy. Observation and prompt intervention are mandatory to deal effectively with cardiac tamponade. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses play important roles in identifying patients at risk for cardiac tamponade and in recognizing signs and symptoms of cardiac tamponade early so this life-threatening emergency can be treated promptly. PMID- 10461702 TI - Disseminated intravascular coagulation. AB - OBJECTIVES: To provide an overview of the pathophysiology, manifestations, diagnosis, and treatment of disseminated intravascular coagulation (DIC) as it occurs in cancer. DATA SOURCES: Published articles, research reports, and book chapters. CONCLUSIONS: The syndrome of DIC is a serious hypercoagulation state that in its acute form may be life-threatening. The hemorrhage and intravascular coagulation that occur with DIC may lead to irreversible morbidity and mortality. Prompt recognition and emergency treatment are necessary to help minimize morbidity and mortality. IMPLICATIONS FOR NURSING PRACTICE: Nurses can play an important role in early recognition of DIC to allow for prompt intervention. Nurses caring for patients affected by DIC will be providing complex nursing care, in addition to psychosocial support to patients and families. PMID- 10461703 TI - Superior vena cava syndrome. AB - OBJECTIVES: To provide a review of the treatment and nursing management of superior vena cava syndrome (SVCS). DATA SOURCES: Review articles, research studies, and book chapters. CONCLUSIONS: SVCS is primarily associated with small cell lung cancer. It usually has a chronic, insidious onset, but may present acutely with laryngeal or cerebral edema. Radiotherapy, chemotherapy, surgery, thrombolysis, and interventional radiology have provided effective treatment. IMPLICATIONS FOR NURSING PRACTICE: Management of the patient with SVCS includes recognition of high-risk patients and initial symptomatology, accurate assessments, appropriate therapies, psychosocial support, and education regarding recurrent SVCS. PMID- 10461704 TI - Hypercalcemia. AB - OBJECTIVES: To provide a review of hypercalcemia of malignancy, including the incidence, pathophysiology, signs and symptoms, treatment, and nursing interventions. DATA SOURCES: Research studies, review articles, proceedings from nursing conferences, and book chapters. CONCLUSIONS: Hypercalcemia of malignancy is difficult to diagnose because the signs and symptoms are similar to those experienced by patients with end-stage cancers. Knowing the cancers and histologic cell types that are at high risk for hypercalcemia assists in early diagnosis and treatment. Treatment decisions must consider potential benefits and their risks and affect on quality of life. IMPLICATIONS FOR NURSING PRACTICE: Hypercalcemia of malignancy as a metabolic oncologic emergency requires accurate assessment and rapid intervention. Early recognition by the patient, family, and health care professionals can result in improved quality of life. PMID- 10461705 TI - Tumor lysis syndrome. AB - OBJECTIVES: To review the pathophysiology, clinical manifestations, and management of tumor lysis syndrome. DATA SOURCES: Published articles, case reports, and book chapters on tumor lysis syndrome. CONCLUSIONS: Tumor lysis syndrome is a very serious and sometimes life-threatening complication of cancer treatment. Identification of patients at risk and initiation of preventative interventions are the focus of medical and nursing management. Ongoing monitoring during and following cancer treatment is necessary to promote early response to changes in patient condition and minimize adverse events. IMPLICATIONS FOR NURSING PRACTICE: Early recognition of signs and symptoms and clinical management of tumor lysis syndrome is a challenging responsibility of the oncology nurse. PMID- 10461706 TI - Sepsis. AB - OBJECTIVES: To review infection and sepsis in patients with cancer and to provide an overview of controversies and research-based practices of infectious complications and management strategies. DATA SOURCES: Research studies, review articles, web sites, and consensus documents. CONCLUSIONS: Traditional assumptions about infection and its optimal management are redefined by research regarding transfusion and catheter-related infections, prophylactic antibiotic administration, use of growth factors, and antimicrobial therapy regimens. IMPLICATIONS FOR NURSING PRACTICE: Infection is still the most common source of morbidity and mortality among cancer patients. The importance of recognizing high risk patients, implementing infection prevention practices, and prompt intervention for infection symptoms has been established. PMID- 10461707 TI - Acute hypersensitivity reactions to chemotherapy. AB - OBJECTIVES: To provide a review of the immune system response involved in acute hypersensitivity reactions (HSRs) and anaphylactic reactions to chemotherapy. DATA SOURCES: Professional experience, published articles, and book chapters. CONCLUSIONS: Acute HSRs from chemotherapy administration can be a rare to moderate occurrence, depending on the chemotherapy agents involved. It is of vital importance for nurses caring for patients receiving chemotherapy to take steps to prevent HSRs when possible and/or to minimize the potentially fatal effects of those reactions. IMPLICATIONS FOR NURSING PRACTICE: Nurses caring for patients receiving chemotherapy should understand the risk of HSRs as well as the pathophysiology involved to ensure utilization of preventive measures when appropriate and early recognition and intervention in the event of an acute HSR. PMID- 10461708 TI - Assessment of sports participation levels following knee injuries. AB - Although there are many published instruments designed to determine outcome following the treatment of knee injuries, only a few incorporate specific assessments of sports activity level and participation into the evaluation. This article reviews 3 of the most commonly used sports activity outcome instruments: the scales devised by Tegner and Lysholm, the Hospital for Special Surgery and the International Knee Documentation Committee. Problems and potential study biases that can arise with improper questionnaire design and data reduction techniques are reviewed, and recommendations are made to correct these problems. The problems identified include: (i) the failure to precisely define sports activity levels according to a specific sport and intensity of participation; (ii) the failure to sort populations according to overall intensity of athletic participation both before and after treatment; (iii) the failure to detect and sort from the population patients who return to sports and experience significant symptoms; (iv) the combination of work and sports activities into the same scale; and (v) the failure to detect alterations in sports participation caused by changes in lifestyle or non-knee-related factors. We have developed a sports assessment instrument, the Cincinnati Sports Activity Scale (CSAS). The methodology used to create this scale, its use in the assessment of return to or change in sports activities, and the assessment of specific functions of daily and sports activities, are briefly reviewed. The CSAS is based on 2 criteria: (i) the frequency of participation; and (ii) the general types of forces experienced by the lower extremity during the sport. The assessment of change in sports activities accounts for modifications in lifestyle and can also detect patients who have returned to sports but are experiencing significant symptoms and problems. Examples of data reduction and reporting are provided to represent practical situations from a prior investigation. PMID- 10461709 TI - Effects of exercise on lipoprotein(a). AB - Lipoprotein(a) [Lp(a)] is a unique lipoprotein complex in the blood. At high levels (> 30 mg/dl), Lp(a) is considered an independent risk factor for cardiovascular diseases. Serum Lp(a) levels are largely genetically determined, remain relatively constant within a given individual, and do not appear to be altered by factors known to influence other lipoproteins (e.g. lipid-lowering drugs, dietary modification and change in body mass). Since regular exercise is associated with favourable changes in lipoproteins in the blood, recent attention has focused on whether serum Lp(a) levels are also influenced by physical activity. Population and cross-sectional studies consistently show a lack of association between serum Lp(a) levels and regular moderate physical activity. Moreover, exercise intervention studies extending from 12 weeks to 4 years indicate that serum Lp(a) levels do not change in response to moderate exercise training, despite improvements in fitness level and other lipoprotein levels in the blood. However, recent studies suggest the possibility that serum Lp(a) levels may increase in response to intense load-bearing exercise training, such as distance running or weight lifting, over several months to years. Cross sectional studies have reported abnormally high serum Lp(a) levels in experienced distance runners and body builders who train for 2 to 3 hours each day. However, the possible confounding influence of racial or ethnic factors in these studies cannot be discounted. Recent intervention studies also suggest that 9 to 12 months of intense exercise training may elevate serum Lp(a) levels. However, these changes are generally modest (10 to 15%) and, in most individuals, serum Lp(a) levels remain within the recommended range. It is unclear whether increased serum Lp(a) levels after intense exercise training are of clinical relevance, and whether certain Lp(a) isoforms are more sensitive to the effects of exercise training. Since elevation of both low density lipoprotein cholesterol (LDL-C) and Lp(a) levels in the blood exerts a synergistic effect on cardiovascular disease risk, attention should focus on changing lifestyle factors to decrease LDL-C (e.g. dietary intervention) and increase high density lipoprotein cholesterol (e.g. exercise) levels in the blood. PMID- 10461710 TI - The future of doping control in athletes. Issues related to blood sampling. AB - When current antidoping programmes were developed, the most frequently used doping agents were xenobiotics, such as stimulants and anabolic steroids, that are readily detectable in urine with the use of gas chromatography and mass spectrometry. As control of traditional doping agents became effective, some athletes turned to other means to improve performance, including blood doping and the application of recombinant peptide hormones such as erythropoietin and growth hormone. Doping with these agents is not easily detected in urine samples, and therefore new strategies must be developed as a supplement to those already in use. Such strategies will probably include analysing blood samples, as several of the most promising methods that are able to detect modern doping agents use blood as the analytical matrix. Non-autologous blood doping results in an admixture of self and foreign red blood cells that can be detected in a blood sample with the methods available. Methods to indicate doping with erythropoietin include the indirect finding of an elevated level of soluble transferrin receptor in serum, or a direct demonstration of a shift from the normal to an abnormal spectrum of erythropoietin isoforms. To indicate doping with growth hormone, a set of serum parameters including insulin growth factors and their binding proteins are under investigation as indirect evidence. A direct method using isotopic differences between endogenous and recombinant growth hormones is being investigated. A similar method has been established to detect the administration of testosterone esters. Several legal and ethical questions must be solved before blood sampling can become a part of routine doping control, but the major ethical question is whether sport can continue as today without proper methods to detect many modern doping agents. PMID- 10461711 TI - Alpine ski bindings and injuries. Current findings. AB - In spite of the fact that the overall incidence of alpine ski injuries has decreased during the last 25 years, the incidence of serious knee sprains usually involving the anterior cruciate ligament (ACL) has risen dramatically since the late 1970s. This trend runs counter to a dramatic reduction in lower leg injuries that began in the early 1970s and to date has lowered the risk of injury below the knee by almost 90%. One of the primary design objectives of modern ski boots and bindings has been to protect the skier from tibia and ankle fractures. So, in that sense, they have done an excellent job. However, despite advances in equipment design, modern ski bindings have not protected the knee from serious ligament trauma. At the present time, we are unaware of any binding design, settings or function that can protect both the knee and lower extremities from serious ligament sprains. No innovative change in binding design appears to be on the horizon that has the potential to reduce the risk of these severe knee injuries. Indeed, only 1 study has demonstrated a means to help reduce this risk of serious knee sprains, and this study involved education of skiers, not ski equipment. Despite the inability of bindings to reduce the risk of severe knee injuries there can be no doubt that improvement in ski bindings has been the most important factor in the marked reduction in incidence of lower leg and ankle injuries during the last 25 years. The authors strongly endorse the application of present International Standards Organisation (ISO) and American Society for Testing and Materials (ASTM) standards concerning mounting, setting and maintaining modern 'state of the art' bindings. PMID- 10461712 TI - Effects of creatine supplementation on exercise performance. AB - While creatine has been known to man since 1835, when a French scientist reported finding this constitutent of meat, its presence in athletics as a performance enhancer is relatively new. Amid claims of increased power and strength, decreased performance time and increased muscle mass, creatine is being hailed as a true ergogenic aid. Creatinine is synthesised from the amino acids glycine, arginine and methionine in the kidneys, liver and pancreas, and is predominantly found in skeletal muscle, where it exists in 2 forms. Approximately 40% is in the free creatine form (Crfree), while the remaining 60% is in the phosphorylated form, creatine phosphate (CP). The daily turnover rate of approximately 2 g per day is equally met via exogenous intake and endogenous synthesis. Although creatine concentration (Cr) is greater in fast twitch muscle fibres, slow twitch fibres have a greater resynthesis capability due to their increased aerobic capacity. There appears to be no significant difference between males and females in Cr, and training does not appear to effect Cr. The 4 roles in which creatine is involved during performance are temporal energy buffering, spatial energy buffering, proton buffering and glycolysis regulation. Creatine supplementation of 20 g per day for at least 3 days has resulted in significant increases in total Cr for some individuals but not others, suggesting that there are 'responders' and 'nonresponders'. These increases in total concentration among responders is greatest in individuals who have the lowest initial total Cr, such as vegetarians. Increased concentrations of both Crfree and CP are believed to aid performance by providing more short term energy, as well as increase the rate of resynthesis during rest intervals. Creatine supplementation does not appear to aid endurance and incremental type exercises, and may even be detrimental. Studies investigating the effects of creatine supplementation on short term, high intensity exercises have reported equivocal results, with approximately equal numbers reporting significant and nonsignificant results. The only side effect associated with creatine supplementation appears to be a small increase in body mass, which is due to either water retention or increased protein synthesis. PMID- 10461713 TI - Popliteal artery entrapment syndrome. AB - Popliteal artery entrapment syndrome (PAES) is a rare cause of exercise-induced leg pain. Entrapment occurs because of an abnormal relationship between the popliteal artery and the surrounding myofascial structures in the popliteal fossa. Arterial insufficiency in the affected limb arises with entrapment of the artery, commonly giving leg symptoms with exertion. The true incidence of PAES in the general population is not known. The aetiology of PAES has an embryological basis related to the development of the popliteal artery and the surrounding musculature. Many different classification schemes have been developed to differentiate the various types of abnormal anatomy that are associated with the syndrome. Repeated popliteal artery compression causes trauma to the arterial wall, leading to premature localised atherosclerosis. The pathology of PAES is believed to be progressive, with arterial thrombosis occurring in some individuals as a natural progression of the disease process. Acute ischaemia can occur if there is an occlusion of the artery or thrombosis within an aneurysm. Clinically, up to 85% of individuals diagnosed with the syndrome are males. The mean age of individuals in a large series was 28 years. The condition can be found bilaterally in 25% of cases. Most individuals present with exercise-induced leg pain, the remainder presenting with acute or chronic ischaemia. The condition can result in significant functional loss for active individuals. Surgery has been advocated to prevent the progression of the disease that is believed to be the natural history of untreated PAES. However, the little research that has been done to determine the prognosis for individuals who have undergone surgery has focused on the patency rate of the arteries after surgery and the presence or absence of complications. Research needs to be done to look at the natural history of untreated PAES and the functional status of athletes after undergoing PAES surgery. PMID- 10461714 TI - Ultrasound technology for hyperthermia. AB - Hyperthermia (HT) is used in the clinical management of cancer and benign disease. Numerous biological and clinical investigations have demonstrated that HT in the 41-45 degrees C range can significantly enhance clinical responses to radiation therapy, and has potential for enhancing other therapies, such as chemotherapy, immunotherapy and gene therapy. Furthermore, high-temperature hyperthermia (greater than 50 degrees C) alone is being used for selective tissue destruction as an alternative to conventional invasive surgery. The degree of thermal enhancement of these therapies is strongly dependent on the ability to localize and maintain therapeutic temperature elevations. Due to the often heterogeneous and dynamic properties of tissues, most notably blood perfusion and the presence of thermally significant blood vessels, therapeutic temperature elevations are difficult to spatially and temporally control during these forms of HT therapy. However, ultrasound technology has significant advantages that allow for a higher degree of spatial and dynamic control of the heating compared to other commonly utilized heating modalities. These advantages include a favorable range of energy penetration characteristics in soft tissue and the ability to shape the energy deposition patterns. Thus, heating systems have been developed for interstitial, intracavitary, or external approaches that utilize properties such as multiple transducer arrays, phased arrays, focused beams, mechanical and/or electrical scanning, dynamic frequency control and transducers of various shapes and sizes. This article provides a general review of a selection of ultrasound hyperthermia systems that are either in clinical use or currently under development, that utilize these advantages as a means to better localize and control HT for the aforementioned therapies. PMID- 10461715 TI - Clinical use of ultrasound tissue harmonic imaging. AB - The recent introduction of tissue harmonic imaging could resolve the problems related to ultrasound in technically difficult patients by providing a marked improvement in image quality. Tissue harmonics are generated during the transmit phase of the pulse-echo cycle, that is, while the transmitted pulse propagates through tissue. Tissue harmonic images are formed by utilizing the harmonic signals that are generated by tissue and by filtering out the fundamental echo signals that are generated by the transmitted acoustic energy. To achieve this, two processes could be used; one by using filters for fundamental and harmonic imaging and the second using two simultaneous pulses with a 180 degrees difference in phase. The introduction of harmonics allows increased penetration without a loss of detail, by obtaining a clearer image at depth with significantly less compromise to the image quality caused by the use of lower frequencies. This imaging mode could be used in different organs with a heightening of low-contrast lesions through artefact reduction, as well as by the induced greater intrinsic contrast sensitivity of the harmonic imaging mode. PMID- 10461716 TI - 3-D ultrasound quantification of neonatal cerebral ventricles in different head positions. AB - We determined the influence of head position on lateral ventricular cerebral volume in low-birth-weight infants by three-dimensional (3-D) ultrasound (US). Thirty-nine neonates were examined prospectively in a controlled and blinded study. We used a freehand 3-D US system to acquire data sets after head positioning for 3 h on left and right side in random order. The borders of the lateral ventricles were marked in stored cross-sections. Volumes were calculated as mean of duplicate measurements. Median volume of lateral cerebral ventricles was 1.03 (quartiles 0.78-1.36) mL. Median left ventricular volume was slightly larger than right one (p = 0.13). Down-side lateral ventricles showed smaller volumes than up-side positioned ventricles (p < 0.01). Freehand 3-D US allows quantification of small volumes as neonatal lateral cerebral ventricles. Head position influences the lateral cerebral ventricle volume in low-birth-weight infants. PMID- 10461717 TI - Iron accumulation in the substantia nigra in rats visualized by ultrasound. AB - In recent studies, we have found a marked increase in substantia nigra (SN) echogenicity in patients with Parkinson's disease (PD) using transcranial ultrasound. Because a substantial body of evidence has accumulated indicating a selective elevation of iron in the SN from patients with PD, we set out to test the hypothesis that trace metals like iron could lead to the observed increase of SN echogenicity in PD. Rat brains were scanned after stereotactic injection of iron in different concentrations into the SN and after injecting ferritin, zinc and 6-OHDA alone, and after the addition of desferrioxamine. The amount of iron in the SN was measured spectroscopically. For iron, and partly for 6-OHDA, in different concentrations, a dose-dependent increase of SN echogenicity could be visualized, corresponding to an increase of iron measured by spectroscopy. No increase of echogenicity was visualized after the injection of ferritin and the addition of desferrioxamine to 6-OHDA, though an increase of iron was measured by spectroscopy. Therefore, we conclude that iron not bound to these proteins may lead to an increase of echogenicity of the SN. PMID- 10461718 TI - Noninvasive 3-D ultrasound of atherosclerotic plaques in the Watanabe rabbit. AB - We have investigated the ability to quantitate atherosclerosis in the aortic arch of the Watanabe rabbit using noninvasive 3-D ultrasound. Our methodology utilizes postprocessing of videotaped freehand 2-D interrogations to form a compound 3-D data block. Structures may then be segmented on the attributed grey-scale level and volumes measured. Analysis of 3-D reconstructions revealed a low echo structure in the aortic arch of atherosclerotic rabbits, absent in nonatherosclerotic rabbits, at recognized sites of plaque predilection. This structure volume correlated closely with fatty streak volume determined from histology (r = 0.890). During a 30-week study, this structure volume increased in untreated animals, but was blocked by treatment with the antiatherosclerotic agent probucol. Thus, a new 3-D ultrasound methodology has been used noninvasively to detect and quantitate a low echo structure corresponding to fatty streaks in the Watanabe rabbit aortic arch. This new methodology could potentially aid plaque burden quantification in human peripheral arteries. PMID- 10461719 TI - Portal venous volume flow: in vivo measurement by time-domain color-velocity imaging. AB - The portal venous velocity and flow volume in 39 patients (16 with liver cirrhosis, 11 with chronic hepatitis, 12 without liver disease) were measured using both color velocity imaging quantification (CVI-Q) and conventional Doppler flowmetry. The average portal venous velocity and flow volume values obtained using the two methods were similar. The correlation coefficients for the paired measurements show positive correlations (velocity: 0.73, p < 0.0001; volume: 0.50, p = 0.001). However, the coefficients of variation between the two methods were not good (velocity: 14.9%, volume: 26.4%). In conventional Doppler flowmetry, the mean velocity to maximum velocity ratio (Vmean:Vmax) is assumed to be constant (Vmean:Vmax = 0.57 in this study). However, the Vmean:Vmax ratios calculated from the flow profile in CVI-Q were 0.67 +/- 0.13 in the patients with liver cirrhosis, 0.58 +/- 0.13 in the patients with chronic hepatitis, and 0.53 +/- 0.08 in the patients without liver disease. Therefore, a measurement method that takes the blood flow profile into account, such as CVI-Q, might be useful for the quantitative measurement of the portal venous velocity and volume. PMID- 10461721 TI - Comparison of transcranial Doppler flow velocity and cerebral blood flow during focal ischemia in rabbits. AB - The predictive value of transcranial Doppler (TCD) cerebral blood flow velocity (CBFV) measurements for cerebral blood flow (CBF) calculations in humans is still controversial, and experimental correlative studies are lacking. The aim of the present study was to validate TCD signals of CBFV during focal cerebral ischemia. Therefore, CBFV determined in the middle cerebral artery (MCA) was compared with values of CBF obtained from autoradiograms of ischemic brain areas. To determine CBFV, a transcranial Doppler ultrasound probe (TCD) adapted to small sample volumes was used in 9 rabbits. CBF was quantified after a final infusion of [14C] iodoantipyrine in the same animals. For focal cerebral ischemia induction, two threads were flushed upward simultaneously into the internal carotid artery, resulting in a flow reduction in the ipsilateral MCA. After thread occlusion, mean systolic CBFV in the MCA decreased from 49 +/- 9 cm/s to 22 +/- 3 cm/s. CBF in the caudate nucleus was reduced (19 +/- 8 mL/100 g/min) compared to the contralateral nonischemic side (52 +/- 18 mL/100 g/min). The decrease in hemispheric CBF correlated well with the decrease in both mean systolic (r = 0.97) and diastolic (r = 0.94) CBFV in the MCA (p < 0.01). The decrease in CBFV determined by transcranial Doppler ultrasound in the MCA appears to reflect the reduction in CBF in the affected brain hemisphere and can be used as a quantitative in vivo parameter for tissue perfusion. PMID- 10461720 TI - Doppler evaluation of the effects of pharmacological treatment of portal hypertension. AB - The splanchnic pharmacodynamic effects of the drugs used for the treatment of hemorrhagic complications of portal hypertension were poorly clarified until some years ago. The introduction of Doppler ultrasound provided a powerful tool to investigate such hemodynamic effects and brought new insights in this field. The present article reviews the pharmacodynamics of the substances used in the treatment of portal hypertension, with particular regard to the effects assessable by duplex Doppler ultrasonography. PMID- 10461722 TI - Fetal transesophageal echocardiography utilizing a 10-F, 10-MHz intravascular ultrasound catheter--comparison with conventional maternal transabdominal fetal echocardiography in sheep. AB - The purpose of our study in fetal sheep was to assess the measurement agreement between fetal transesophageal echocardiography (FTEE) and conventional maternal transabdominal echocardiography (CMTFE) by the Bland-Altman method. We performed our study in 11 fetal sheep between 95-103 days of gestation (term = 145 days). FTEE was performed by imaging the fetal heart in horizontal planes utilizing a 10 F, 10-MHz intravascular ultrasound catheter. CMTFE was carried out using a 5.0 MHz phased-array transducer replicating the FTEE imaging planes. We found close agreement between FTEE and CMTFE measurements of great vessel and cardiac valvar dimensions. Conversely, the variability between both techniques for measuring ventricular dimensions was inadequate. We conclude that FTEE permits measurement of great vessel and cardiac valve dimensions with high agreement with CMTFE measurements. This finding strengthens the applicability of FTEE as a monitoring tool during experimental open or fetoscopic fetal cardiac interventions. PMID- 10461723 TI - 3-D power Doppler cerebral angiography in neonates and young infants: comparison with 2-D power Doppler angiography. AB - The aims of this study were to evaluate the ability of 3-dimensional (3-D) power Doppler angiography (3DPDA) to depict the intracranial vasculature in infants, to compare with 2-D power Doppler ultrasonography (2DPDU), and to explore the potential clinical applications of this procedure in young infants with brain disorders. We performed 3DPDA in 27 infants. 2DPDU were completed in both sagittal and coronal directions in 12 of these patients. In the other 15, only right sagittal plane images were available for comparison. Using a grading system and with only vessels with more than half of the length demonstrated included for comparison, we compared the Doppler signals of major vessels. 3DPDA could have good visualization in more than 60% of the internal carotid artery, ophthalmic artery, pericallosal artery, callosomarginal artery, internal cerebral vein, vein of Galen, and straight sinus in the sagittal plane. 3DPDA also could have good demonstration in about 50% of basilar artery in coronal plane, and posterior communicating artery, posterior cerebral artery, and lenticulostriate artery in sagittal plane. 3DPDA was better than 2DPDU in demonstrating all the major intracranial vessels in different planes, except the anterior communicating artery. In the anterior communicating artery, neither can demonstrate more than 30%. PMID- 10461724 TI - The use of the wavelet transform to describe embolic signals. AB - A number of methods to detect cerebral emboli and differentiate them from artefacts using Doppler ultrasound have been described in the literature. In most, Fourier transform-based (FT) spectral analysis has been used. The FT is not ideally suited to analysis of short-duration embolic signals due to an inherent trade-off between temporal and frequency resolution. An alternative approach that might be expected to describe embolic signals well is the wavelet transform. Wavelets are ideally suited for the analysis of sudden short-duration signal changes. Therefore, we have implemented a wavelet-based analysis and compared the results of this with a conventional FFT-based analysis. The temporal resolution, as measured by the half-width maximum, was significantly better for the continuous wavelet transform (CWT), mean (SD) 8.40 (8.82) ms, compared with the 128-point FFT, 12.92 (9.70) ms, and 64-point FFT, 10.80 (5.69) ms. Time localization of the CWT for the embolic signal was also significantly better than the FFT. The wavelet transform appears well suited to the analysis of embolic signals offering superior time resolution and time localization to the FFT. PMID- 10461725 TI - Frequency analysis of echo texture in tendon. AB - A texture discriminant based on spatial frequencies is proposed for characterizing B-scans of Achilles' tendon. The anisotropic echo texture of normal tendon has an ellipsoidal spatial spectrum that can be quantified by the ratio of the major-to-minor axis and by the direction of the major axis. Applying a moving window to the B-scan, a corresponding tissue elliptical axis ratio (TEAR) image is derived that segments out tendon. The algorithm was applied to B scan images taken from 13 volunteers, 6 of whom had tendon abnormalities: tendon rupture (n = 3) or cholesterol deposits (xanthomas) in patients with heterozygous familial hypercholesterolemia (n = 3). The average TEAR value was 1.75 +/- 0.17 for normal tendon, 1.04 +/- 0.06 for torn tendon, and 1.31 +/- 0.16 for tendons with xanthomas. The dispersion of the directionality vectors was used to further differentiate tendons with xanthomas from normal tendons. This technique appears to be useful for characterizing both diffuse and focal tendon abnormalities. PMID- 10461726 TI - The axolotl as an animal model for the comparison of 3-D ultrasound with plain film radiography. AB - We assessed the usefulness of an animal model, the axolotl (Ambystoma mexicanum), in comparing 3-D ultrasound (3-D US) and plain film radiographs. Hindlimbs were amputated from 5 animals, at either the zeugopodial or stylopodial level, and each regenerating limb was imaged 16 times with 3-D US and 14 times with plain film X ray over 315 days. US images were acquired with a Siemens Sonoline Versa Pro and a 10-MHz linear array transducer. For 3-D US images, the probe was translated in a motor-driven linear stage while images were digitized. The regenerating tibia and fibula bones were detected on 3-D US an average of 37 days earlier than on plain film radiography, and regenerating phalangeal bones were detected on 3-D US an average of 18 days earlier. After 120 days, both imaging modalities consistently showed the bones. The average bone growth rates for the tibia and fibula were 0.019 +/- 0.001 mm/day and 0.019 +/- 0.001 mm/day, respectively. PMID- 10461727 TI - Liquid jets, accelerated thrombolysis: a study for revascularization of cerebral embolism. AB - A prior study has reported that a rapid recanalization therapy of cerebral embolism, using liquid jet impacts generated by the interaction of gas bubbles with shock waves, can potentially penetrate through thrombi in as little as a few microseconds with very efficient ablation (Kodama et al. 1997). The present study was undertaken to examine the liquid jet impact effect on fibrinolysis in a tube model of an internal carotid artery. First, the conditions for generating the maximum penetration depth of liquid jets in the tube were investigated. Gelatin was used to mimic thrombi. The shock wave was generated by detonating a silver azide pellet weighing about a few micrograms located in a balloon catheter. The collapse of the inserted gas bubbles and the subsequent liquid jet formation were recorded with high-speed photography. Second, thrombi were formed using fresh human blood from healthy volunteers. The fibrinolysis induced by the liquid jet impact with urokinase was explored. This was conducted under selected conditions based on the experiment using the gelatin. Fibrinolysis was calculated as the percentage of the weight loss of the thrombus. Fibrinolysis with urokinase alone and with a single liquid jet impact with urokinase was 1.9 +/- 3.7% (n = 16) and 20.0 +/- 9.0% (n = 35), respectively, for an incubation time of 60 min. Statistical differences were obtained between all groups (ANOVA). These results suggest that liquid jet impact thrombolysis has the potential to be a rapid and effective therapeutic modality in recanalization therapy for patients with cerebral embolism and other clinical conditions of intra-arterial thrombosis. PMID- 10461728 TI - Hemostasis of punctured vessels using Doppler-guided high-intensity ultrasound. AB - The use of Doppler ultrasound was investigated to determine if it would aid in guiding the application of high-intensity focused ultrasound (HIFU) to stop bleeding from punctured vessels. Major vessels (abdominal aorta, illiac, carotid, common femoral and superficial femoral arteries and the jugular vein) were surgically exposed, punctured and treated in anesthetized pigs. Treatment was applied when the Doppler sounds indicated the focus coincided with the bleeding site. In 89 treatment trials, the average time to achieve major hemostasis (a point where bleeding was reduced to a level of only oozing) was 8 s, and for complete hemostasis was 13 s. These times were significantly shorter than those of an identical former study in which only visual guidance was used. In that study, the average times for major and complete hemostasis were 40 and 62 s, respectively. The advantage of Doppler guidance in applying HIFU in treating bleeding vessels was demonstrated. PMID- 10461729 TI - Effect of high-intensity focused ultrasound on whole blood with and without microbubble contrast agent. AB - Using human whole blood samples with and without contrast agent (CA), we evaluated the effect of exposures to focused, continuous wave (CW) 1.1-MHz ultrasound for durations of 10 ms to 1 s at spatial average intensities of 560 to 2360 W/cm2. Cavitation was monitored with a passive cavitation detector and hemolysis was determined with spectroscopy. In whole blood alone, no significant cavitation, heating or hemolysis was detected at any exposure condition. Conversely, cavitation and hemolysis, but not heating, were detected in whole blood with CA. A CA concentration as low as 0.28 microL CA per mL whole blood at an intensity of 2360 W/cm2 for 1 s resulted in measurable cavitation and a 6-fold increase in hemolysis compared to shams. Cavitation and hemolysis increased proportional to the concentration of CA and duration of exposure. In samples containing 4.2 microL CA per mL whole blood exposed for 1 s, a threshold was seen at 1750 W/cm2 where cavitation and hemolysis increased 10-fold compared to exposures at lower intensities. HIFU exposure of whole blood containing CA leads to significant hemolysis in vitro and may lead to clinically significant hemolysis in vivo. PMID- 10461730 TI - Transient poration and cell surface receptor removal from human lymphocytes in vitro by 1 MHz ultrasound. AB - The study objective was to gain insight into ultrasound-induced, sub-lytic cell surface modifications. Two primary hypotheses were tested by flow cytometric methods; viz., sonication will: 1. remove all or part of a specific cell surface marker in lymphocytes surviving insonation, and 2. induce transient pores in the cell membranes of some surviving cells. RPMI 1788 human lymphocytes were exposed in vitro to 1-MHz, continuous-wave ultrasound (approximately 8 W/cm2 ISP) for 30 s, which lysed approximately 50% of the cells. Insonation: 1. altered cell morphology, increasing the population of cells of reduced size but high structure (designated as population R2), many of which were nonviable, and diminishing the population of cells of large size and high structure (designated as population R1), most of which were viable, 2. diminished the fluorescence signal from the pan B lymphocyte marker CD19 in populations R1 and R2 to equivalent extents, and 3. increased by approximately 7-fold the number of transiently permeabilized cells in R1, as evidenced by simultaneous uptake of propidium iodide and fluorescein diacetate. The results indicate that ultrasound-induced CD19 removal from R1 cells can occur without accompanying gross membrane loss. The cell morphology/mortality shifts indicate that the ultrasound-induced morphological change is associated with lethal membrane poration, suggesting that the diminished CD19 fluorescence signal from insonated R2 cells arises partly by simultaneous loss of membrane fragments, CD19 and cytoplasm. PMID- 10461731 TI - Acoustic saturation and output regulation. AB - Acoustic saturation pressures are predicted for ultrasonic beams of a range of frequencies and focal depths. Using reasonable approximations, saturation values for mechanical index (MI) and derated spatial-peak, time-average and pulse average intensities are calculated. These are compared with thresholds set for regulatory purposes by the U. S. Food and Drug Administration (FDA), and by the International Electrotechnical Commission (IEC). It is concluded that there are many conditions for which acoustic saturation in water prevents the values of MI and regulated intensities from exceeding thresholds set by the FDA. These conditions are particularly associated with higher frequencies and deeper focal lengths. The thresholds for action set by IEC 61157 are sufficiently low that similar problems do not arise. It is concluded that present regulations are not fully effective in limiting the output from diagnostic ultrasound equipment, and that some conditions exist that are not subject to output control. PMID- 10461732 TI - Date-based exports: MAFF explains how the scheme will work. PMID- 10461733 TI - Current British veterinary attitudes to perioperative analgesia for dogs. AB - In March 1996, a questionnaire was sent to 2000 veterinary surgeons, primarily involved in small animal practice, to assess their attitudes to perioperative analgesic therapy in dogs, cats and other small mammals. This paper is concerned only with the data relating to dogs. The veterinary surgeons considered that pain was a consequence of all the surgical procedures specified, but there were differences in their treatment of pain. Some veterinarians considered that a degree of pain was necessary postoperatively to prevent excessive activity. In general, women and more recent graduates assigned higher pain scores to the procedures and were more likely to treat the pain with analgesics. A significant number of veterinarians consider the use of opiates or non-steroidal anti inflammatory drugs before surgical procedures, but relatively few appear to use combinations of different classes of analgesics either before or after operations. PMID- 10461734 TI - Effects of glucose infusion on the endocrine, metabolic and cardiorespiratory responses to halothane anaesthesia of ponies. AB - Glucose was infused intravenously into six ponies during halothane anaesthesia, to evaluate its effect on their endocrine response to anaesthesia. The ponies were premedicated with acepromazine, and anaesthesia was induced with thiopentone and maintained with halothane in oxygen for two hours. Glucose was infused to maintain the plasma glucose concentration above 20 mmol/litre. Anaesthesia was associated with hypothermia, a decrease in haematocrit, hypotension, hyperoxaemia, respiratory acidosis and an increase in the plasma concentrations of lactate and arginine vasopressin. The concentration of beta-endorphin in plasma increased transiently after 20 minutes but there were no changes in concentrations of adrenocorticotrophic hormone, dynorphin, cortisol or catecholamines. These data suggest that the glucose infusion attenuated the normal adrenal response of ponies to halothane anaesthesia. PMID- 10461735 TI - Hydronephrosis and ureteral duplication in a young alpaca. AB - A seven-month-old male alpaca (Lama pacos) with signs of abdominal straining was examined. A fluid-filled structure was palpable in the mid-abdominal region, and ultrasonography revealed a hydronephrosis of the right kidney, with an associated mega-ureter. The affected kidney was removed and the clinical signs resolved. Histological examination of the kidney revealed the unusual congenital abnormality of ureteral duplication. It is suggested that the hydronephrosis developed as a result of this underlying condition. PMID- 10461736 TI - Increased S-100b in the cerebrospinal fluid of some cattle with bovine spongiform encephalopathy. PMID- 10461737 TI - Comparison of ventrodorsal and dorsoventral radiographic projections for hip dysplasia diagnosis. PMID- 10461738 TI - Outbreak of acute ovine mastitis associated with Pseudomonas aeruginosa infection. PMID- 10461739 TI - S-100 and bovine spongiform encephalopathy. PMID- 10461740 TI - False acacia poisoning in horses. PMID- 10461741 TI - Hairworms of birds of prey. PMID- 10461742 TI - Rhabditis dermatitis in sheep in Saudi Arabia. PMID- 10461743 TI - A novel D-leucine-containing Conus peptide: diverse conformational dynamics in the contryphan family. AB - A Conus peptide family (the contryphans) is noteworthy because of the presence of a post-translationally modified D-amino acid in all members of the family. A new contryphan peptide, Leu-contryphan-P, was isolated from the venom of Conus purpurascens; the sequence of this peptide is: Gly-Cys-Val-D-Leu-Leu-Pro-Trp-Cys OH. This is the first known occurrence of D-leucine in a Conus peptide. The discovery of Leu-contryphan-P suggests that there may be branches of the contryphan peptide family that diverge much more in sequence than previously anticipated. Several natural contryphans provide dramatic examples of interconversion between multiple conformational states in small constrained peptides. The contryphans that have 4-trans-hydroxyproline and D-tryptophan in positions 3 and 4, respectively, exhibit two peaks under reverse-phase HPLC conditions, indicating interconversion between two discrete conformations. However, [L-Trp4]contryphan-Sm (with L-Trp substituted for D-Trp) exhibits a single, broad peak that elutes later than the natural peptide, suggesting that D Trp stabilizes a conformation in which hydrophobic residues are buried. Leucontryphan-P which has valine and D-leucine instead of 4-trans-hydroxyproline and D-tryptophan shows only a single peak that elutes much later than the other contryphans. PMID- 10461744 TI - Fluoroalcohols as structure modifiers in peptides and proteins: hexafluoroacetone hydrate stabilizes a helical conformation of melittin at low pH. AB - The effect of hexafluoroacetone hydrate (HFA) on the structure of the honey bee venom peptide melittin has been investigated. In aqueous solution at low pH melittin is predominantly unstructured. Addition of HFA at pH approximately 2.0 induces a structural transition from the unstructured state to a predominantly helical conformation as suggested by intense diagnostic far UV CD bands. The structural transition is highly cooperative and complete at 3.6 M (50% v/v) HFA. A similar structural transition is also observed in 2,2,2 trifluoroethanol which is complete only at a cosolvent concentration of approximately 8 M. Temperature dependent CD experiments support a 'cold denaturation' of melittin at low concentrations of HFA, suggesting that selective solvation of peptide by HFA is mediated by hydrophobic interactions. NMR studies in 3.6 M HFA establish a well defined helical structure of melittin at low pH, as suggested by the presence of strong NH/NHi+1 NOEs throughout the sequence, along with many medium range helical NOEs. Structure calculations using NOE-driven distance constraints reveal a well-ordered helical fold with a relatively flexible segment around residues T10-G11-T12. The helical structure of melittin obtained at 3.6 M HFA at low pH is similar to those determined in methanolic solution and perdeuterated dodecylphosphocholine micelles. HFA as a cosolvent facilitates helix formation even in the highly charged C-terminal segment. PMID- 10461745 TI - Synthesis and kinetics of cyclization of MHC class II-derived cyclic peptide vaccine for diabetes. AB - Conformationally constrained cyclic peptides are known to be better vaccines because of their ability to mimic the native structure of a protein against which an immune response is sought. To test the hypothesis of using conformationally constrained, disease-associated, MHC-derived peptides as vaccines for the prevention of type I diabetes, a 22 amino acid nonobese diabetic(NOD) mouse MHC class II-derived synthetic peptide was cyclized by the formation of end-to-end disulfide bonds and used to prevent diabetes and insulitis in NOD mice. The peptide was synthesized by Fmoc chemistry and cyclized using two methods: a commercially available cyclizing resin (Ekathiox) and air oxidation. When a 10 m excess of resin was used, the Ekathiox yielded a substantial amount of cyclic peptide with few or no side reactions. The kinetics of cyclization by air oxidation at different temperatures indicated that increasing both temperature and pH decreased the cyclization time significantly. Air oxidation at pH 10 at 37 55 degrees C yielded the desired product within 2 h. PMID- 10461746 TI - Development of a photoreactive parathyroid hormone antagonist to probe antagonist receptor bimolecular interaction. AB - Parathyroid hormone (PTH) and PTH-related protein (PTHrP) exert their calciotropic activities by binding to a specific seven-transmembrane-helix containing G protein-coupled receptor mainly located in bone and kidney cells. In order to map in detail the nature of hormone-receptor interaction, we are employing 'photoaffinity scanning' of the bimolecular interface. To this end, we have developed photoreactive benzophenone (BP)-containing PTH analogs which can be specifically and efficiently cross-linked to the human (h) PTH/PTHrP receptor. In this report, we describe the photocross-linking of a BP-containing PTH antagonist, [Nle8,18,D-2-Nal12,Lys13(epsilon-BP),2-Nal23,Tyr34]bPT H(7-34)NH2 (ANT) to the recombinant hPTH/PTHrP receptor stably expressed in human embryonic kidney cells (HEK-293, clone C-21). This photoreactive antagonist has high affinity for the hPTH/PTHrP receptor and inhibits agonist-induced cyclase activity and intracellular calcium release. The photo-induced cross-linking of the radioiodinated antagonist (125I-ANT) to the recombinant hPTH/PTHrP receptor followed by SDS-PAGE analysis reveals a single radiolabeled band of approximately 85kDa, similar to that observed after cross-linking of a radioiodinated BP containing agonist. The formation of this covalent 125I-ANT - hPTH/PTHrP receptor conjugate is competed dose-dependently by a variety of unlabelled PTH- and PTHrP derived agonists and antagonists. This is the first report of a specific and efficient photocross-linking of a radioiodinated PTH antagonist to the hPTH/PTHrP receptor. Therefore, it provides the opportunity to study directly the nature of the bimolecular interaction of PTH antagonist with the hPTH/PTHrP receptor. PMID- 10461747 TI - Design, synthesis and characterization of antimicrobial pseudopeptides corresponding to membrane-active peptide. AB - To obtain active and metabolically stable analogues, peptide backbone modifications have been incorporated into many biologically active peptides. In this study, we designed and synthesized pseudopeptides corresponding to the antimicrobial peptide that acted on the lipid membrane of the pathogen. Most pseudopeptides exhibited a longer half-life than the peptide in the presence of serum as well as a considerable activity against test bacteria and fungi. Circular dichroism spectra and retention times of the pseudopeptides helped us to elucidate the effect of the incorporation of backbone modifications on the structural parameters necessary for the activity, indicating that alpha-helical structure was the most important factor for the activity and hydrophobicity had a considerable effect on the activity. Backbone modifications employed in this study can be a useful tool for structure-activity relationship studies and the development of therapeutic agents from membrane-active antimicrobial peptides. PMID- 10461748 TI - The solution structure of uperin 3.6, an antibiotic peptide from the granular dorsal glands of the Australian toadlet, Uperoleia mjobergii. AB - Uperin 3.6 (GVIDA5AKKVV10NVLKN15LF-NH2) is a wide-spectrum antibiotic peptide isolated from the Australian toadlet, Uperoleia mjobergii. With only 17 amino acid residues, it is smaller than most other wide-spectrum antibiotic peptides isolated from amphibians. In 50% (by vol.) trifluoroethanol, an NMR study and structure calculations indicate that uperin 3.6 adopts a well-defined amphipathic alpha-helix with distinct hydrophilic and hydrophobic faces. Examination of the activities of synthetic modifications of uperin 3.6 reveal that the three lysine residues are essential for antibiotic activity. PMID- 10461749 TI - Synthesis and cytostatic activities of didemnin derivatives. AB - The highly cytostatic didemnins contain a 23-membered cyclopeptolide with a side chain attached to the backbone through the amine group of threonine. Thirty-six derivatives varying the side chain were prepared, but only compounds with D-MeLeu attached to threonine show remarkable biological activities. To protect the macrocycle from degradation by lipases the two ester bonds were replaced successively by amide bonds. Although these variations have a major effect on the conformation and rigidity of the ring, the compound which contains exclusively amide bonds is highly active, equivalent to acetyl-didemnin A. PMID- 10461750 TI - Effects of amounts of additives on peptide coupling mediated by a water-soluble carbodiimide in alcohols. AB - The optimal amounts of 1-hydroxybenzotriazole (HOBt), 3,4-dihydro-3-hydroxy-4-oxo 1,2,3-benzotriazine (HOOBt) and 1-hydroxy-7-azabenzotriazole (HOAt) for enhancement of peptide coupling mediated by 1-ethyl-3-(3 dimethylaminopropyl)carbodiimide (EDC) hydrochloride in alcoholic solvents were found to be less than equimolar against the carboxyl component or the carbodiimide. In comparison with the use of equimolar additives, the use of less equimolar ones was more effective in suppressing the competitive ester formation and in increasing the yield of desired peptides. EDC hydrochloride/around 0.1 equimolar HOAt or HOOBt were efficient reagents for peptide synthesis in the media. PMID- 10461751 TI - Convenient preparation of [Orn(Tfa)2]- and [Orn(Boc)2, Orn(Tfa)2]gramicidin S, versatile unsymmetrically protected derivatives of gramicidin S. AB - Treatment of gramicidin S (GS) with trifluoroacetic anhydride afforded a derivative in which only one of the two Orn side chains was trifluoroacetylated in 72% yield, furnishing the first efficient method for the preparation of a monoprotected derivative of GS. The mono(Tfa) derivative [Orn(Tfa)2']GS was treated with di-tert-butyl dicarbonate to yield dually protected derivative [Orn(Boc)2,Orn(Tfa)2']GS from which another monoprotected derivative [Orn(Boc)2]GS was prepared in high yield. These unsymmetrically protected GS derivatives are versatile starting materials for the preparation of various other GS derivatives. As an example of application of the unsymmetrically protected derivatives, a dimeric GS derivative was prepared via a singly p nitrobenzenesulfonyl(NBS)-activated derivative [Orn(Boc)2,Orn(NBS)2']GS. PMID- 10461752 TI - International radiobiology archives of long-term animal studies: structure, possible uses and potential extension. AB - Animal experiments have contributed a great deal to our information on effects and risks arising from exposure to radionuclides. This applies, in particular, to alpha-emitting radionuclides where information from man is limited to thorotrast, 224Ra and 226Ra. The late C.W. Mays was the first to suggest that animal data in conjunction with epidemiological data could allow estimates of human risks for radionuclides - predominantly from actinides - where information in man is scarce. The 'International Radiobiology Archives of Long-term Animal Studies' were created through the combined efforts of European, American and Japanese scientists and aim to safeguard the large amount of existing data on long-term animal experiments and make them available for, among others, an improved assessment of risks from alpha-emitting radionuclides. This paper summarizes the structure of the archives and reviews their present status and future plans. It also demonstrates the extensive information available in these archives on alpha emitting radionuclides which is suitable for further analysis. Also, the structure of the animal archives could - in a slightly modified form - accommodate the epidemiological data available on 224Ra and thorotrast and, thus, facilitate a direct comparison of data from man, dogs and rodents. PMID- 10461753 TI - Applications of amorphous track models in radiation biology. AB - The average or amorphous track model uses the response of a system to gamma-rays and the radial distribution of dose about an ion's path to describe survival and other cellular endpoints from proton, heavy ion, and neutron irradiation. This model has been used for over 30 years to successfully fit many radiobiology data sets. We review several extensions of this approach that address objections to the original model, and consider applications of interest in radiobiology and space radiation risk assessment. In the light of present views of important cellular targets, the role of target size as manifested through the relative contributions from ion-kill (intra-track) and gamma-kill (inter-track) remains a critical question in understanding the success of the amorphous track model. Several variations of the amorphous model are discussed, including ones that consider the radial distribution of event-sizes rather than average electron dose, damage clusters rather than multiple targets, and a role for repair or damage processing. PMID- 10461754 TI - The Bethe surface of liquid water. AB - The Bethe surface of liquid water, which was previously calculated by using a semiempirical model, is compared with recently available data from IXS experiments (inelastic x-ray scattering; Compton scattering of high energy photons) in liquid water. No alarming discrepancy is found for a global view of the Bethe surface, in part because the two sets of data have been constrained by the Bethe sum rule. The shape of the Bethe ridge given by the new data is broader than that obtained through the impulse approximation. The extrapolation to the optical limit (viz., at zero momentum transfer) is shown, and the reliability of these data is discussed in detail. PMID- 10461755 TI - A Monte Carlo code for positive ion track simulation. AB - An ion interaction model has been described for simulating positive ion tracks in a variety of media with the capability of interfacing with several secondary electron transport codes. Data are presented for single-and double-differential cross-sections, binding energies, probability density distribution for delocalisation parameters for conductors and tissue, branching ratios and ionisation efficiencies for water vapour and liquid water. PMID- 10461756 TI - Formation of single- and double-strand breaks of pBR322 plasmid irradiated in the presence of scavengers. AB - By the method of gel electrophoresis, radiation-induced DNA single- and double strand breaks (SSB, DSB) were studied with a model system of pBR322 solution in vitro in the presence of 'OH radical scavengers, mannitol and TE (10(-2) mol dm( 3) Tris and 10(-3) mol dm(-3) ethylene diamine tetra-acetic acid). Experiments showed that SSB resulted from one-hit events of radiation energy deposition and DSB resulted from both one-hit and two-hit energy deposition events and so were distinguished into two classes of alphaDSB and betaDSB. Moreover, alpha/beta, where alpha is the number of DSB per unit dose induced in one irradiation event and beta the number of DSB per unit squared dose induced by the combination of two independent SSB, was related to the scavenging capacity, sigma, and for sigma > 10(8) s(-1), alphaDSB predominate over DSB. On the other hand, if sigma<2x10(8)s(-1), the measured G(alphaDSB) decreased in parallel with G(SSB), i.e., G(alphaDSB)/G(SSB) was a constant. When sigma>2x10(8) s(-1). G(alphaDSB) decreased slightly so that the ratio of alphaDSB to SSB evidently increased. Therefore, alphaDSB could be induced by the radical transfer mechanism for sigma<2x10(8) s(-1) and contrarily produced by the local multiply damaged sites (LMDS) mechanism for sigma larger than this value. In addition, the distance for two independent complementary SSB forming a DSB was deduced, but no apparent variation of it was found in the wide sigma range from approximately 10(5) to approximately 10(9) s(-)1, which shows that the DNA steric structure was not influenced by mannitol. PMID- 10461757 TI - Biological effects of ion beams in Nicotiana tabacum L. AB - The biological effects of ion beams on Nicotiana tabacum L., particularly the induction of chromosome aberrations, were investigated. Dry seeds were exposed to 12C5+, 4He2+ and 1H+ beams with linear energy transfer (LET) ranging from 1 to 111 keV/microm and irradiated with gamma-rays. Ion beams were more effective in reducing germination and survival of the seeds than gamma-rays. The LD50 for 12C5+ beams, 4He2+ beams and gamma-rays were 35, 60 and 500 Gy, respectively. The frequencies of mitotic cells with chromosome aberrations, such as chromosome bridges, acentric fragments and lagging chromosomes in the root tip cells of the exposed seeds, increased linearly with increasing doses. Relative biological effectiveness (RBE) values, based on the doses that induced a survival inhibition of 50% and a 10% frequency of aberrant cells, were 14.3-17.5 for the 12C5+ beams, 7.0-8.3 for the 4He2+ beams and 7.8 for the 1H+ beams. Furthermore, the relative ratios of the chromosome aberration types were significantly different between the ion beam and the gamma-ray regimes: chromosome fragments were more frequent in the former, and chromosome bridges in the latter. Based on these results, we concluded that the repair process of initial le PMID- 10461758 TI - Reduction of radiocaesium transfer to broiler chicken meat by a clinoptilolite modified with hexacyanoferrate. AB - The effect of RADEKONT (a natural clinoptilolite modified by hexacyanoferrate) on 137Cs uptake into meat was tested in experiments with broiler chickens. Three experiments determined the influence of RADEKONT on radiocaesium transfer after single or repeated applications of artificially contaminated feed mixture and one experiment investigated the effect of RADEKONT when feeding a mixture containing wheat contaminated by the Chernobyl fallout. Independent of the effect of RADEKONT, the uptake of radiocaesium was faster in leg meat than in breast meat. Reduction factors (137Cs transfer without the RADEKONT additive compared with those observed after supplementation of the additive into the feed mixture) of 1.1-1.3 and 1.2-2.3, respectively, were achieved after single and repeated administrations of artificially contaminated feed. No significant differences in reduction between breast and leg meat were observed. RADEKONT was more effective when the chickens were fed with Chernobyl-contaminated wheat (reduction factors of up to 3.7) than an artificial 137Cs source. RADEKONT as a supplement during the decontamination period decreased the biological half-life of 137Cs to less than 1 day. The timing of the application of RADEKONT might be important in determining its effectiveness, especially in young, rapidly growing chickens. PMID- 10461759 TI - Nuclear translocation of the p65 subunit of NF-kappaB in L5178Y sublines differing in antioxidant defense. AB - We examined the induction of nuclear translocation of the p65 subunit of NF kappaB in L5178Y (LY) cells. We used two LY sublines which are inversely cross sensitive to hydrogen peroxide and x-rays: LY-R cells are radioresistant and oxidant-sensitive, whereas LY-S cells are radiosensitive and oxidant-resistant. Hydrogen peroxide, phorbol ester and x-rays caused a marked translocation of p65 NF-kappaB in LY-R cells and a weak translocation in LY-S cells. By manipulating the antioxidant defense status, we obtained an alteration in the p65-NF-kappaB translocation induction in LY-R cells. A similar effect was achieved with lovastatin pretreatment (25 microM, 24 h, 37 degrees C). The response of LY-S cells under all these conditions was considerably weaker. We conclude that differential nuclear translocation of p65-NF-kappaB in LY sublines is not related to the lethal effect of the activating, damaging agent; rather it is due to the more efficient antioxidant defense in LY-S than in LY-R cells. PMID- 10461760 TI - Repair of cellular radiation damage in space under microgravity conditions. AB - The influence of microgravity on the repair of x-ray-induced DNA double-strand breaks was studied in the temperature-conditional repair mutant rad54-3 of diploid yeast Saccharomyces cerevisiae. Cells were exposed on the ground and kept at a low temperature until microgravity conditions were achieved. In orbit, they were incubated at the permissive temperature to allow repair. Before re-entry they were again cooled down and kept at a low temperature until final analysis. The experiment, which was flown on the shuttle Atlantis on flight STS-76 (SMM 03), showed that repair of pre-formed DNA double-strand breaks in yeast is not impaired by microgravity. PMID- 10461761 TI - Enhanced airborne radioactivity during a pine pollen release episode. AB - A single episode of pine pollen release in the highly contaminated area of Novozybkov, Russian Federation, which led to enhanced atmospheric concentrations of 137Cs is discussed. The pollen grains were sampled by a rotating arm impactor and analysed by gamma-spectrometry for 137Cs activity and by image analysis for their size. In the vicinity of a forest, a maximum concentration of 4.5+/-0.4 mBq m(-3) was measured, and a mean activity per pollen grain of 260+/-80 nBq was determined. The emission rate of the Novozybkov mixed pine forest was estimated to be approximately 400 Bq m(-2) per year. Because of the large size of pine pollen grains (about 50 microm) and the short emission period of 5-8 days per year, the estimated potential annual inhalation doses are very low. Biological emissions including pollen release may be a source of increased airborne radionuclide concentrations at larger distances from the source areas as well. PMID- 10461762 TI - Mechanism underlying the therapeutic effects of electroconvulsive therapy (ECT) on depression. AB - Electroconvulsive therapy (ECT) is used to treat drug-resistant depressive disorders. The results of studies on the mechanism underlying the effectiveness of ECT on depression are still controversial. ECT stimulus is usually larger than the threshold of induction of seizures and activation of whole-brain is believed to be necessary to produce therapeutic effects. A single ECT session induces alterations of the electroencephalogram (EEG) including initial epileptic discharges, then slow waves, and finally flattened EEG. Repeated ECT results in an increasing number of slower waves in the EEG for as long as a month. ECT induced changes in various neurotransmitter systems have also been reported. Serotonin (5-hydroxytryptamine, 5-HT) is one of the most important neurotransmitters involved in depressive illness, and ECT alters several 5-HT receptor subtypes in the central nervous system. 5-HT1A receptors in post synaptic neurons are sensitized by repeated ECT, but those in pre-synaptic neurons (auto-receptors) are not changed. In addition, our electrophysiological studies have shown that ECT increases sensitivity to 5-HT of 5-HT3 receptors in the hippocampus, resulting in an increase in release of neurotransmitters such as glutamate and gamma-aminobutyric acid. In contrast, ECT decreases the auto receptor functions in noradrenergic and dopaminergic neurons in the locus coeruleus and substantia nigra, respectively, resulting in an increase in release of noradrenaline and dopamine. In conclusion, 5-HT1A-receptor sensitization may be important for explaining the effectiveness of ECT, as this change induces a decrease in the number of 5-HT2A receptors that are elevated in depressive patients. Facilitation of neurotransmitter releases due to 5-HT3-receptor sensitization by ECT may also play an important role in effective treatment of depressive patients refractory to therapeutic drugs. PMID- 10461763 TI - Ca2+ signaling and the insulin secretory cascade in the pancreatic beta-cell. AB - Recent progress in electrophysiological and microscopic techniques have enabled us to estimate exocytotic and pre-exocytotic events in the secretory machinery in single pancreatic beta-cells. We have been studying mechanisms involved in the regulation of insulin granule movement, which supplies release-ready granules, by direct visualization of granule traffic in living beta-cells and found the movement to be regulated by a mechanism different from that controlling exocytosis. From the obtained findings together with those from electrophysiological approaches, a new understanding of the role of the crucial second messenger Ca2+, and other second messengers, as well as resultant protein phosphorylation has been generated. The aim of this review is to describe a synergistic network for the control of insulin release by second messengers and protein kinases. PMID- 10461764 TI - Therapeutic effects of a calcium antagonist, lacidipine, on stroke-prone spontaneously hypertensive rats with cerebrovascular lesions. AB - The aim of this study was to investigate the therapeutic effectiveness of lacidipine in stroke-prone spontaneously hypertensive rat (SHRSP) with cerebrovascular lesions in comparison with nicardipine. SHRSP were fed 1% saline as drinking water. After the onset of stroke, saline was replaced with water and each drug was administered orally once a day for 3 weeks. In the drug-untreated group, recurrence of stroke was repeated, deterioration and amelioration of neurological deficits (ND) were repeated, and histological examination and measurement of regional blood flow (rBF) using nonradioactive colored microspheres performed at the end of treatment revealed severe damages and significantly decreased rBF in brain and kidney, respectively. In kidney, not only lacidipine (1 mg/kg) but also nicardipine (30 mg/kg) decreased vascular lesions and ameliorated low-rBF significantly. Both drugs also inhibited the recurrence of stroke completely even at a low dose that did not ameliorate severe hypertension. Neuronal damages and ND in each lacidipine-treated group were ameliorated significantly, whereas those in each nicardipine-treated group were slightly improved. Lacidipine at 1 mg/kg alone ameliorated the cerebral low-rBF significantly even at 24 hr after administration. These results suggest that a long-lasting improvement of low-rBF after stroke may be useful in the treatment of SHRSP with cerebrovascular lesions. PMID- 10461765 TI - A comparison of the antagonistic activities of tamsulosin and terazosin against human vascular alpha1-adrenoceptors. AB - Tamsulosin, a selective alpha1A-adrenoceptor antagonist, and terazosin, a non selective one, are effective for the treatment of urinary disturbance due to benign prostatic hypertrophy. In the present study, their alpha1-adrenoceptor blocking effects on blood vessels, which may cause orthostatic hypotension, were investigated in 10 healthy males. After the subjects took orally 0.2 mg of tamsulosin, 1 mg of terazosin or a lactate capsule as the control in a randomized cross-over fashion, their finger tip vasoconstrictor response to cold stimulation and vasoconstrictor response of the dorsal hand vein to increasing doses of phenylephrine were examined. The finger tip vasoconstrictor response was significantly reduced and the infusion rate of phenylephrine producing a half maximal constriction was significantly increased by terazosin, but tamsulosin had no significant effect on these parameters. These data suggest that the usual dose of tamsulosin exerts little alpha1-adrenoceptor-blocking activity on blood vessels, and orthostatic episodes might be mild, if any, during the treatment with tamsulosin. PMID- 10461766 TI - Protective effect of histidine on hydroxyl radical generation induced by potassium-depolarization in rat myocardium. AB - We investigated the efficacy of histidine on potassium-depolarization induced hydroxyl radical (*OH) generation in the extracellular fluid of rat myocardium by a flexibly mounted microdialysis technique (O system). After the rat was anesthetized, a microdialysis probe was implanted in the left ventricular myocardium, and then sodium salicylate in Ringer's solution (0.5 nmol/microl per minute) was infused to detect the generation of *OH as reflected by the nonenzymatic formation of 2,3-dihydroxybenzoic acid (DHBA). Infusion of KCl (70 mM) clearly produced an increase in *OH formation. However, when KCl in the presence of a high concentration of histidine (25 mM) was infused through the microdialysis probe, KCl failed to increase the 2,3-DHBA formation. To examine the effect of histidine on ischemia-reperfusion of the myocardium, the heart was subjected to myocardial ischemia for 15 min by occlusion of the left anterior descending coronary artery (LAD). When the heart was reperfused, a marked elevation of the levels of 2,3-DHBA was observed in the heart dialysate. However, when corresponding experiments were performed with histidine (25 mM)-pretreated animals, histidine prevented the ischemia-reperfusion induced *OH formation trapped as 2,3-DHBA. These results indicate that histidine may protect against K+ depolarization-evoked *OH generation in rat myocardium. PMID- 10461767 TI - No participation of adenosine A1 receptor in acute nephrotoxicity by 4-pentenoic acid administration in dogs. AB - Intrarenal infusion of 4-pentenoic acid is known to lower renal cortical ATP content and cause a reduction in glomerular filtration rate (GFR). The alteration in nucleotide metabolism might augment the production of adenosine, thereby eliciting the fall in GFR. This study was conducted to examine whether 4 pentenoic acid stimulates renal production of adenosine, and if so, to examine the role of adenosine A1 receptor in the reduction of GFR by 4-pentenoic acid. With infusion of 4-pentenoic acid (1 micromol x kg(-1) x min(-1)) into the renal artery of anesthetized dogs, GFR gradually decreased and reached minimum at 60 min with values ranging from 33.9+/-2.2 to 20.2+/-2.8 ml/min. Neither renal blood flow nor mean arterial pressure was affected, but tubular reabsorption of water and sodium was significantly attenuated. Renal venous plasma concentration and urinary excretion of adenosine rose markedly (20-fold) without any change in arterial concentration, suggesting that renal adenosine production was augmented by 4-pentenoic acid. However, KW-3902 (8-(noradamantan-3-yl)-1,3 dipropylxanthine), a selective antagonist of the adenosine A1 receptor, did not affect the action of 4-pentenoic acid on GFR or renal handling of water and sodium. It is concluded that 4-pentenoic acid markedly increases renal adenosine production, but adenosine A1 receptor is not involved in the 4-pentenoic acid induced nephrotoxicity. PMID- 10461769 TI - Inhibition by alkylamines of NADPH oxidase through blocking the assembly of enzyme components. AB - Alkylamines inhibit NADPH oxidase both in intact neutrophils and in a cell-free system. The aim of this study was to examine the mechanism underlying this inhibitory effect. Among alkylamines with different chain lengths, the C12 compound (laurylamine) showed the greatest inhibitory effect on the cell-free NADPH oxidase activity induced by arachidonic acid (AA) in the presence of GTPgammaS. The inhibition was overcome by further addition of AA, and it was observed irrespective of whether laurylamine was added before or after the enzyme activation by AA. When added prior to the enzyme activation, laurylamine blocked translocation to the membrane of all three cytosolic components (p47-phox, p67 phox and rac) in a cell-free translocation assay. When added after the activation, laurylamine released only rac from the membrane. Laurylamine did not inhibit the reduction of cytochrome c by xanthine oxidase, suggesting that it does not have superoxide-scavenging activity. These results indicate that laurylamine inhibits both the activation process of NADPH oxidase and the activated enzyme itself by blocking the assembly of the oxidase components. PMID- 10461768 TI - Antiulcer effect of lafutidine on indomethacin-induced gastric antral ulcers in refed rats. AB - Lafutidine is a new type antiulcer agent with antisecretory and gastroprotective activities. We investigated the effect of lafutidine on indomethacin-induced antral ulcer in refed rats. Subcutaneous indomethacin injection resulted in the formation of gastric antral ulcer. Lafutidine (1-10 mg/kg, p.o.) reduced the area of ulcer in a dose-dependent manner when administered immediately after the indomethacin injection. Capsaicin at 3 mg/kg, p.o. and 16,16-dimethyl prostaglandin E2 at 3 microg/kg, p.o. also reduced the ulcer area. Chemical deafferentation of capsaicin-sensitive neurons or N(G)-nitro-L-arginine treatment aggravated the ulcer formation and abolished the preventive effect of lafutidine and capsaicin. After the induction of gastric ulcer, lafutidine given twice daily for 2.5 days reduced the area of ulcer in a dose-dependent manner with a significant effect at 10 mg/kg, p.o., as compared with that of the control group. In chemically-deafferentated rats, lafutidine did not show any healing effect. Cimetidine (30 mg/kg, p.o.) and famotidine (1 mg/kg, p.o.) had no significant effect on indomethacin-induced antral ulcer. These results may suggest that lafutidine, unlike cimetidine and famotidine, can prevent the indomethacin induced antral ulcer formation and accelerate the healing of the ulcer in refed rats through mechanisms involving the capsaicin-sensitive afferent neurons and nitric oxide. PMID- 10461770 TI - Effects of K+ channel modulators on the relationship between action potential duration and Ca2+ transients in single ventricular myocytes of the guinea pig. AB - Effects of K+ channel modulators, cromakalim and E4031 [1-[2-(6-methyl-2-pyridyl) ethyl]-4-(4-methylsulfonylaminobenzoyl) piperidine], on the relationship between the action potential duration (APD) and Ca2+ transients were examined in single myocytes isolated from guinea pig cardiac left ventricle. Application of cromakalim decreased APD at 90% repolarization (APD90) and Ca2+ transient elicited at 0.5 Hz (IC50s=0.6 and 3 microM, respectively). Application of 0.3 microM E4031 increased these parameters by 110% and 45%, respectively. Under voltage-clamp, the relation between the duration of depolarization to 0 mV and Ca2+ transients could be described by the sum of two exponential components; the time constants were approximately 5 and 280 msec, respectively. The first component was abolished by 10 microM ryanodine, suggesting the involvement of Ca2+-induced Ca2+ release (CICR). Neither cromakalim nor E4031 directly affected Ca2+ current and Ca2+ transients under voltage clamp. When APD was changed by K+ channel modulators, the relation between APD90 and Ca2+-transients was almost similar to that obtained by changing the depolarization duration under voltage clamp. CICR was changed significantly only when APD90 was markedly shortened by cromakalim. The extensively prolonged AP and Ca2+ transient in the presence of E4031 were reduced by an addition of cromakalim. It is concluded that these two K+ channel modulators can significantly alter the AP-induced Ca2+ transient mainly by changing APD, which regulates both Ca2+ influx and extrusion. PMID- 10461771 TI - Histamine release induced by immobilization, gentle handling and decapitation from mast cells and its inhibition by nedocromil in rats. AB - The effect of immobilization, gentle handling and decapitation on the level of plasma histamine in Wistar rats was investigated. Mast cell deficient (Ws/Ws) rats were used to characterize the source of elevated histamine in plasma by stress, and the effect of nedocromil, a mast cell stabilizer, on histamine release was assessed in these models in vivo. The plasma histamine concentration of freely moving rats was 93.0+/-2.3 pmol/ml. Gentle handling produced a transient increase in plasma histamine level by 1.9-fold, whereas immobilization resulted in a longer-lasting elevation by 2.6-fold compared to that in the freely moving rats. Decapitation increased the plasma histamine level by 10- to 16-fold compared with that in the freely moving rats. No increase in plasma histamine was found in Ws/Ws rats exposed to stress. Nedocromil inhibited the increase in plasma histamine level induced by stress in a dose-dependent manner. These findings suggest that stress induces histamine release from mast cells in Wistar rats and the extent of this histamine release increases with the severity of stress. Nedocromil proved to be a good pharmacological tool to inhibit stress induced release of mediators from mast cells. PMID- 10461772 TI - Pharmacological evaluation of a new laser blood flowmeter for measuring coronary blood flow, assessed in the canine isolated, blood-perfused ventricular tissue preparation in comparison with an electromagnetic flowmeter. AB - A new portable-type laser blood flowmeter was recently developed for measuring the blood flow in vessels. The sensitivity and specificity of the laser flowmeter was assessed in comparison with the well-established electromagnetic flowmeter using a canine isolated, blood-perfused ventricular tissue preparation. The laser flowmeter can record the phasic pattern of the coronary blood flow like the electromagnetic flowmeter. The extent of the changes after intracoronary administration of ACh and angiotensin II as well as coronary occlusion was almost identical between these two methods. These results suggest that the new laser flowmeter may possess potential utilities in both basic experimental and clinical practices. PMID- 10461773 TI - Chronotropic, inotropic, dromotropic and coronary vasodilator effects of bisaramil, a new class I antiarrhythmic drug, assessed using canine isolated, blood-perfused heart preparations. AB - The cardiovascular effects of a new class I antiarrhythmic drug, bisaramil, were examined using canine isolated, blood-perfused heart preparations. Bisaramil exerted negative chronotropic, inotropic and dromotropic effects as well as coronary vasodilator action, which are qualitatively the same as those of classical class I drugs. The selectivity of bisaramil for the intraventricular conduction vs the other cardiac variables was compared with that of disopyramide and flecainide. Bisaramil was the most selective for intraventricular conduction, while it was the least selective for ventricular muscle contraction. We conclude that bisaramil may become a useful antiarrhythmic drug with less cardiac adverse effects. PMID- 10461774 TI - Effects of phenylephrine and prazosin on axial movement of the rat incisor and arterial blood pressure. AB - We investigated the dose-response effects of phenylephrine and antagonistic effects of prazosin on axial movement of the rat incisor and arterial blood pressure. Phenylephrine caused a temporal extrusive tooth movement and an increase in blood pressure at all doses. With increasing phenylephrine doses, the maximum extrusive tooth movement and maximum increase in blood pressure were enhanced. The maximum extrusive tooth movement and increase in blood pressure induced by phenylephrine were markedly suppressed after pretreatment with prazosin. These results suggested that extrusive tooth movement is closely related to the rise in blood pressure due to stimulation of vascular alpha1 receptors. PMID- 10461775 TI - The Edwin Smith Surgical Papyrus. AB - The Edwin Smith Surgical Papyrus is undoubtedly one of the most significant medical texts ever discovered. It is of particular interest to neurosurgeons because of its specific references to ancient neurosurgical cases and is the first written record of many terms of neurosurgical interest. This review describes the colorful and controversial history of the Edwin Smith Papyrus and gives translations of four cases of neurosurgical interest. PMID- 10461776 TI - Endoscopic approach to arachnoid cyst. AB - A prospective study of 36 consecutive patients with congenital arachnoid cysts treated endoscopically is reported. There were 15 female and 21 male patients. The mean age at the time of diagnosis was 12.3 years (10 days to 38 years). Arachnoid cysts were located in the suprasellar region in 16 patients, the sylvian fissure in 11, the quadrigeminal cistern in 4 and the posterior fossa in 5. Endoscopic fenestration was combined with cysto-peritoneal shunting for 6 temporal cysts and with ventriculo-peritoneal shunting in 1 suprasellar cyst. Mean postoperative follow-up was 4.2 years (range 1-8 years). Follow-up imaging studies showed that 28 arachnoid cysts (77.8%) were obliterated after endoscopic procedures. Long-term clinical results were good in all patients, although the cysts of 8 patients were not reduced in size. There was no mortality or morbidity. We conclude that endoscopic procedures may be a promising alternative to microsurgical operations or shunting for the treatment of arachnoid cysts. PMID- 10461777 TI - Intracranial hemorrhage in an infant owing to vitamin K deficiency despite prophylaxis. AB - A 3-month-old male infant with intracranial hemorrhage attributable to a vitamin K deficiency is reported. Vitamin K2 was administered orally at birth and then at 5 days and I month of age. Oral antibiotics were also given 2 days before the onset of bleeding. Although the incidence of intracranial hemorrhage resulting from vitamin K deficiency has decreased since the introduction of vitamin K2 prophylaxis, spontaneous intracranial hemorrhages are still being reported in infants. We suggest that vitamin K prophylaxis is needed especially for breast fed infants and for those undergoing antibiotic therapy. PMID- 10461778 TI - The different forms of neurofibromatosis. AB - In the last two decades our knowledge of the natural history, genetics and management of the different forms of neurofibromatosis has changed. Of the numerical classifications of neurofibromatosis proposed in the past, only neurofibromatosis type 1 (Nf1) and neurofibromatosis type 2 (Nf2) have been shown to be distinct at clinical and molecular levels. Mosaicism has been demonstrated both in patients with Nf1 and in patients with Nf2, and features of segmental or mosaic Nf1 and Nf2 have been defined. The outlying phenotypes and the molecular genetics of other, rarer, types of neurofibromatosis have been delineated: these are hereditary spinal neurofibromatosis, Schwannomatosis, familial intestinal neurofibromatosis, autosomal dominant "cafe-au-lait spots alone", autosomal dominant "neurofibromas alone", Watson syndrome, Noonan/neurofibromatosis syndrome and the so-called syndrome of multiple naevi, multiple schwannomas and multiple vaginal leiomyomas. In this article I will review the different forms of neurofibromatosis, focusing on those aspects that most commonly challenge the neurosurgeon. PMID- 10461779 TI - Neurosurgical management of brain abscesses in children. AB - The authors review the management of brain abscesses (BAbs) in 59 pediatric cases. The major surgical procedure used to treat them was repeated puncture and aspiration (51 cases, or 86.44%), excision (8 cases - 13.56%) being of secondary importance. The main etiology was metastatic lesions (24 cases - 40.68%), those due to cyanotic congenital heart disease being the most frequent. Single lesions dominated (41 cases - 69.49%). Gram-positive cocci were the main bacteria involved (31 cases, 52.5%). CT scan represented the main tool in the diagnosis and follow-up. Puncture and aspiration in BAbs led to a significant decrease in mortality (7 cases - 11.86%), and the incidence of seizures and neurological deficits was also reduced. Mortality was significantly correlated with the following factors: consciousness status, multiple BAbs location and hematogenous dissemination. There were recurrences in 11 cases (18.64%), all observed after aspiration procedures. The current concepts for complex management of BAbs, as reflected in recent literature data, are reviewed, with particular emphasis on the use of real-time CT or ultrasound-guided operative techniques. PMID- 10461780 TI - International Multicenter Study of Head Injury in Children. ISHIP Group. AB - With the object of evaluating different epidemiological factors in the acute phase of head injury (HI) in the pediatric age group in five countries (Argentina, Brazil, France, Hong-Kong and Spain), we carried out a prospective and descriptive study, in which we analyzed the clinical and radiological risk factors versus management and outcome 7-30 days after trauma. We included all children seen in the emergency department and hospitalized who were aged between 0 and 15 years and had sustained HI. Data were compiled from the clinical records and analyzed for neurological evaluation with the Glasgow Coma Scale (GCS) and the Glasgow Paediatric Coma Score (GPCS), and also by means of dynamics, symptoms, skull X-rays, CT scans. The total of 2478 patients enrolled in the study was made up of 60.9% boys and 39.1 % girls. Age distribution was as follows: 55.2% aged 0-4 years; 28.3% aged 5-9 years, and 16.4% aged 10-15 years. Most (75.3%, or 1768) of these patients completed follow-up. The total sample included 1058 children (42.7%) who required hospitalization. Skull fractures were identified in 11.8% (298) of the cases, and 6.4% (158) of CT scans were pathologic. Minor HI accounted for 56.4% of these children, moderate HI for 38.9%, and severe HI for the remaining 4.7%. The lethality rate was 1.6%. Our preliminary data reveal that it is very important for new guidelines on the treatment of minor HI to be prepared, because patients with minor HI had undergone the most skull X-rays and also most frequently been admitted to hospital for unnecessarily long periods of time, though the incidence of brain damage (1.6%) was lowest in this group of the study population. We intend to carry out a full analysis of the various risk factors at the end of the study. PMID- 10461781 TI - Fatal head injury in children: a new approach to scoring axonal and vascular damage. AB - As part of a multidisciplinary study of brain damage in children fatally injured in motor vehicle accidents, a simple method to quantify and visualise the distribution and extent of injury has been developed. Vascular and axonal injury were assessed using coronal brain sections stained for haematoxylin and eosin, or reacted immunohistochemically for beta-amyloid precursor protein. Subsequent analysis was carried out using NIH Image software, and the resulting information is displayed in schematic diagrams. These summary diagrams simply and clearly show the distribution of injury in both the coronal and horizontal planes. This technique offers an advantage over previous scoring methods in that it provides both a quantitative and a visual summary of the distribution and extent of brain injury. This information can then be used to compare the injury distribution and severity with estimated impact points and acceleration data. PMID- 10461782 TI - Cephaloceles and abnormal venous drainage. AB - Seven cases of parietal cephalocele and three cases of occipital cephalocele associated with abnormal venous drainage were evaluated, and the mechanism of the development for venous system was discussed from the embryological viewpoint. In parietal cephaloceles the abnormally draining vein, which corresponds to the straight sinus, ascended along the interhemisphere apart from the tentorium and emptied into the superior sagittal sinus below the cephalocele. The superior sagittal sinus formed the confluence, which was positioned high up. Intraoperative findings revealed that cephalocele penetrated the superior sagittal sinus at the midline. In the case of occipital cephalocele, the straight sinus followed a postero-superior course, to drain into the confluence just above the neck of cephalocele. The results of our study suggest that the causation of abnormal venous drainage in the great vein of Galen, the straight sinus and the superior sagittal sinus may be secondary, through interaction with a developmentally pre-existing cephalocele. PMID- 10461783 TI - Neurenteric cyst: its various presentations. AB - Neurenteric (NE) cyst is an uncommon developmental lesion lined with epithelium of endodermal origin. To investigate the clinical manifestation and response to surgery, we retrospectively analyzed eight cases of NE cyst that has been confirmed by surgery. Four were in children. The duration of follow-up ranged from 2 to 105 (mean 38) months. One cyst was in the ventral portion of the posterior cranial fossa and the other seven were on the spinal cord. The chief complaints were motor weakness (5), pain (2), and voiding difficulty (1). In one child and three adults, the duration of symptoms was more than 3 years. Children tended to show rapid progression and excellent recovery after surgery. Although total removal of cyst was possible only in two cases, there was no recurrence. The presentation of an NE cyst may be insidious. Clinical suspicion is important for an early diagnosis and better outcome. Because of the benign course after subtotal excision, too-aggressive removal of the lesion should be avoided. PMID- 10461784 TI - Ethical issues in managed and rationed care for children with severe neurological disabilities: a questionnaire survey. AB - The attitudes of pediatric neurosurgeons to managed and rationed care for children with severe al neurological disabilities were surveyed as reflected in responses from International Society for Pediatric Neurosurgery (ISPN) members to a questionnaire. Of 399 ISPN members, 156 (39.1%) responded to the questionnaire. There were 15 questions, which were designed to explore what care is medically indicated, whether all medically indicated care should always be provided, and how this care should be managed or rationed. Most respondents agreed that these patients should receive the same level of medical care as a normally developing child. However, respondents felt that cardiopulmonary resuscitation (CPR) is not indicated if a child is not retrievable. Most respondents also felt that provision of care should be influenced by cost. The responses to the questions from ISPN members were compared with those from Child Neurology Society (CNS) members. PMID- 10461785 TI - A case of malignant tumour of the jugular foramen in a young infant. AB - Malignant tumours of jugular foramen are very rare in infants. To our knowledge there have been no earlier reports. The case of an infant who had a malignant schwannoma in the region of the jugular foramen is now reported. The difficulties of identifying a particular nerve as the origin of such a tumour are also indicated. PMID- 10461786 TI - Cavernous hemangioma of the skull in a neonate. AB - Cavernous hemangiomas rarely occur in the calvarium and most commonly present in middle-age. Although a congenital vascular disorder can theoretically cause a diploic lesion in any age group, a calvarial cavernous hemangioma has not been reported in newborn. A 4-month-old male infant presented with a large left parietal mass that had been present since birth. Total resection was performed. Pathological examination revealed a cavernous hemangioma developing within the diploic space adjacent to prior hemorrhages. Surgery was performed in this case because of the size and persistence of the lesion. Removal of tumors of a benign nature from the calvarium can be done safely. Cavernous hemangioma of the skull in a neonate should be considered as one of the differential diagnoses in the case of suspected ossified cephalohematoma. PMID- 10461787 TI - Aneurysm of the posterior inferior cerebellar artery caused by a traumatic perforating artery tear-out mechanism in a child. AB - Traumatic posterior circulation aneurysms in the absence of fractures and penetrating wounds are extremely uncommon, especially in children. To our knowledge this is the first traumatic posterior inferior cerebellar artery(PICA) aneurysm reported that cannot be related to a skull fracture or a trauma caused by the edge of a rigid meningeal structure. In the present case, the initial subarachnoid hemorrhage (SAH) was caused by a perforating artery, originating from the PICA, which was torn out as the result of a deceleration trauma. Such a mechanism explains both the initial SAH and the development of the false aneurysm responsible for the second SAH. PMID- 10461788 TI - Gunshot wound of the atlas in a youth. AB - The case of a bullet retained, without causing neurological symptoms, in the anterior arch of a youth's atlas after a gun had been fired a short distance from his mouth is reported. The patient was managed with external stabilization. PMID- 10461789 TI - Giant posterior fossa teratoma. AB - In this study we report a rare case of a giant midline posterior fossa teratoma; its clinical presentation, radiological appearance, treatment and outcome, with an extensive review of the literature. PMID- 10461790 TI - Dynamic delay time compensation for sampling capillaries used in respiratory mass spectrometry. AB - In intensive care patients who receive ventilatory support or full mechanical ventilation, valuable information can be drawn from gas exchange measurements. In this setting, the most favorable method for gas exchange measurement is by simultaneous recording of gas concentrations and gas flow, and by time resolved multiplication and accumulation. This paper presents a new method to compensate for the signal delay time which occurs when a sampling capillary is used for measuring gas concentrations with a respiratory mass spectrometer or some equivalent sidestream gas analyzer. The signal delay of gas concentrations must be accurately compensated to avoid error accumulation in gas exchange calculation. A delay time can be easily measured with a test gas in a laboratory setup and be readily compensated for during the measurements in a ventilated patient. This is a standard procedure which gives reasonable results under normal conditions. Special attention is however required in cases where the gas viscosity changes due to large changes in gas composition, e.g., those used for diagnostic breathing or ventilatory maneuvers. Such changes of viscosity will influence the delay time of the capillary, because they affect its flow resistance. As a consequence they will degrade the quality of measurements when done with a simple fixed delay compensation. The method described here consists of an algorithm which enables compensation for such a temporally changing delay time due to changes in gas composition. PMID- 10461791 TI - The use of PC based VR in clinical medicine: the VREPAR projects. AB - Virtual reality (VR) is an emerging technology that alters the way individuals interact with computers: a 3D computer-generated environment in which a person can move about and interact as if he actually was inside it. Given to the high computational power required to create virtual environments, these are usually developed on expensive high-end workstations. However, the significant advances in PC hardware that have been made over the last three years, are making PC-based VR a possible solution for clinical assessment and therapy. VREPAR - Virtual Reality Environments for Psychoneurophysiological Assessment and Rehabilitation - are two European Community funded projects (Telematics for health - HC 1053/HC 1055 - http://www.psicologia.net) that are trying to develop a modular PC-based virtual reality system for the medical market. The paper describes the rationale of the developed modules and the preliminary results obtained. PMID- 10461792 TI - Biotransport in the human respiratory system. AB - The human respiratory system is an 'open' organ, which is designed to exchange oxygen and carbon dioxide between the circulating blood and the external environment. This gas exchange is successfully accomplished via a set of transport phenomena comprised of oscillatory air flow, heat and water vapor exchange, mucus transport and air-blood gas exchange all of which take place in a complex geometry that undergoes large changes. These transport phenomena occur simultaneously to supply the body's need for oxygen in different physiological conditions and/or environments, while defending it from external hazards. The need for better comprehension of the mechanisms involved in pulmonary diseases and for advanced techniques for both diagnosis and intervention stimulated numerous studies of the different biotransport processes that take place in the human respiratory system. PMID- 10461793 TI - Data compression methods for EEG. AB - Modem brain research experiments require the recording of large amounts of high accuracy EEG data. Sampling frequency, bit resolution and number of channels are significantly larger than in routine clinical measurements. Because of this, the need of efficient signal compression has emerged. This paper presents a survey of available methods and explores some techniques, both lossy and lossless, for compressing EEG signals. PMID- 10461794 TI - Problems associated with FES-standing in paraplegia. AB - Prolonged immobilization, such as occurs after the spinal cord injury (SCI), results in several physiological problems. It has been demonstrated that the standing posture can ameliorate many of these problems. Standing exercise can be efficiently performed by the help of functional electrical stimulation (FES). The first application of FES to a paraplegic patient was reported by Kantrowitz in 1963. It was later shown by our group that standing for therapeutic purposes can be achieved by a minimum of two channels of FES delivered to both knee extensors. The properties of the stimulated knee extensors (maximal isometric joint torque, fatiguing, and spasticity) were not found as sufficient conditions for efficient standing exercise. According to our studies, the ankle joint torque during standing is the only parameter which is well correlated to the duration of FES assisted standing. For good standing low values of the ankle joint torque are required. To minimize the ankle joint torque the lever belonging to the vertical reaction force must be decreased. Adequate alignment of the posture appears to be the prerequisite for efficient FES assisted and arm supported standing exercise. Some patients are able to assume such posture by themselves, while many must be aided by additional measures. At present, surface stimulation of knee extensors combined with some appropriately "compliant shoes" looks to be adequate choice. PMID- 10461795 TI - The Traveling Shutter Wave analyses non-linear compliance during mechanical ventilation. AB - Mechanical ventilation is an important, often life-saving component of modern intensive care medicine. However, it may further aggravate pulmonary pathology by endinspiratory overdistension of the alveoli or by their endexpiratory collapse. To prevent both the ventilator may be adjusted based on the slope of the pressure volume curve, named as compliance, which is often determined by a stepwise inflation of the lungs. This maneuver gained no widespread clinical acceptance because of being cumbersome and invasive. Therefore, we developed a modification of the well known interrupter technique - the Traveling Shutter Wave. A wave of short-term (300 ms) occlusions "travels" over the tidal volume range. Differential compliance is calculated by division of volume and pressure differences between two adjacent occlusion maneuvers. The technique is well suited for the clinical setting because the ventilatory pattern does not need to be changed. This manuscript describes the realization of the Traveling Shutter Wave as well as its application in two patients. PMID- 10461796 TI - A comparison of the plasma fructose concentrations in dogs and cats and changes in the fructose concentrations in dogs following intravenous administration of fructose. AB - The plasma concentrations of fructose, glucose, free fatty acids (FFA) and triglycerides (TG) were measured in dogs and cats. Changes in these concentrations were investigated in dogs by an intravenous fructose tolerance test (IVFTT) at a dose of 0.1 g/kg body weight. Fructose concentrations in the plasma of dogs were significantly higher than those of cats. There was no significant difference in plasma glucose concentrations between dogs and cats. Plasma FFA concentrations decreased and TG concentrations increased after feeding in both dogs and cats. During the IVFTT, the plasma fructose concentrations in the dogs increased rapidly to a peak by 2 min and then decreased to half of the peak by 5 min after the administration of fructose. Administration of fructose resulted in an increase in the plasma TG concentrations and reduced plasma FFA concentrations in the dogs. Only 4%, of the administered fructose was detected in the urine of dogs following IVFTT. Plasma fructose was considered to be rapidly absorbed and metabolized in both dogs and cats. However, as with glucose metabolism, there appear to be some differences in fructose metabolism between dogs and cats. PMID- 10461797 TI - Prevalence of anti-Toxoplasma gondii antibody in wild boar, Sus scrofa riukiuanus, on Iriomote Island, Japan. PMID- 10461798 TI - Hepatic lipidosis associated with cobalt deficiency in Omani goats. AB - Livers from 36 of 684 (5.3%) apparently healthy goats examined at an abattoir in the greater Muscat area of Oman exhibited gross pathological findings characterized by extremely pale, friable, fatty livers encompassing the entire organ. Histopathologically, diffuse hepatic lipidosis and occasional bile duct proliferation were observed. Periodic acid Schiff-positive, diastase-resistant pigment was observed in the macrophages lining the sinusoids. These histopathological lesions were consistent with those characteristic of ovine white liver disease. Cobalt analysis revealed that normal livers had six times more cobalt and a 3-fold less fat content than those measured in the fatty livers. This is the first report of an association between cobalt deficiency and hepatic lipidosis in Omani goats. PMID- 10461799 TI - Lack of sex-influence on the in vitro metabolism of ivermectin by hepatic microsomal preparations from cattle. PMID- 10461801 TI - Studies on normal haematological and serum biochemical values of the 'Hijin' racing camels (Camelus dromedarius) in Kuwait. AB - Haematological estimations and serum biochemical analyses were made on 100 samples collected from clinically healthy 'Hijin' racing camels (Camelus dromedarius) in Kuwait. The red blood cell counts, packed cell volume, haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration and total white blood cell counts were estimated. In the serum biochemical analyses, sodium, potassium, iron, calcium, phosphorus, magnesium, total bilirubin, blood urea nitrogen, creatinine, glucose, total protein and cholesterol concentrations were measured, as were the alanine aminotransferase, aspartate aminotransferase, gamma-glutamyltransferase, creatine kinase and alkaline phosphatase activities. The results are discussed in relation to other findings reported in camels. PMID- 10461800 TI - Influence of diet type and pretreatment fasting on the disposition kinetics of albendazole in sheep. AB - The influence of the quality and quantity of diets on the disposition kinetics of albendazole were studied in sheep in two different experiments. The plasma concentration profiles of albendazole sulphoxide and albendazole sulphone were measured following intraruminal administration of albendazole at 5.0 mg/ kg body weight in weaner sheep offered three different diets: 100% green Sorghum spp., 100% dry mature Cenchrus ciliaris hay and a 50:50 mix of these two diets. The peak plasma concentrations and the availability of the albendazole metabolites, as measured by the area under the concentration time curve, were significantly higher (p < 0.01) in the animals offered exclusively dry fodder compared to other diets. Changing the diet from dry to green fodder resulted in a significantly lower systemic availability of the drug metabolites. It is suggested that a decreased transit time of the digesta in the bowel on the green diet, with its high water content, limited the systemic availability of the drug by reducing the time available for gastrointestinal absorption. An experiment on the influence of different levels of pretreatment fasting on the pharmacokinetics of albendazole revealed significantly higher (p < 0.05) plasma concentrations of the anthelmintically active sulphoxide metabolite from 12 h onwards following administration of the drug in animals subjected to 24 h of pretreatment fasting compared to other groups with pretreatment fasting of 8, 12 or 18 h. The area under the concentration time curve and the minimum residence time of the drug metabolites were significantly greater (p < 0.05) in animals that had been fasted for 24 h. It is suggested that fasting induces a decrease in the flow of digesta through the gastrointestinal tract of ruminants and prolongs the duration of dissolution of the drug, resulting in enhancement of the absorption of albendazole and of the systemic availability of its metabolites. PMID- 10461802 TI - Random amplification of polymorphic DNA fingerprinting of Trypansoma evansi. AB - The total genomic DNAs from Trypanosoma evansi isolates of bubaline, equine and cameline origin were amplified by polymerase chain reaction using eight arbitrarily selected 10-base primers. Informative band patterns were obtained for all isolates analysed. Depending upon the T. evansi isolate primer combination, between 1 and 11 reproducible DNA fingerprints of 205 to 3016 bp were amplified, suggesting minor and major differences in their RAPD (random amplified polymorphic DNA) profiles. PMID- 10461803 TI - The effects on reproductive performance in the short and medium term of the combined use of exogenous melatonin and progestagen pessaries in ewes with a short seasonal anoestrous period. AB - The effects on reproductive performance in the short and medium term of the combined use of exogenous melatonin and progestagen pessaries in ewes with a short seasonal anoestrous period. PMID- 10461804 TI - Spatio-temporal localization of the face inversion effect: an event-related potentials study. AB - Event-related potentials (ERPs) from 58 electrodes at standard EEG sites were recorded while 14 subjects performed a delayed-matching task on normal and inverted faces. A large and single difference between normal and inverted face processing was observed at occipito-temporal sites about 160 ms following stimulus onset, mainly in the right hemisphere (RH). Although the topographies indicate that similar areas are involved at this latency in processing the two types of stimuli, the electrophysiological activity, which corresponds to the previously described N170, was larger and delayed for inverted as compared to normal face processing. These results complement and specify, at a neural level, previous behavioral and divided visual field studies which have suggested that the loss of configural face information by inversion may slow down and increase the difficulty of face processing, particularly in the RH. PMID- 10461805 TI - Effects of CS-US interval modification on diminution of the unconditioned response in electrodermal classical conditioning. AB - The purpose of this experiment was to study whether interstimulus interval (ISI) modification differentially affects the amplitude of the unconditioned response (UR amp.). Seventy-five volunteer subjects received discrimination training with an interval between conditioned stimulus (CS) and unconditioned stimulus (US; aversive white noise) of 8 s. After the discrimination training phase, subjects exhibiting discriminative control were randomized into three groups that differed according to the ISI, of 1, 5 or 8 s. The subjects of each group were then tested with five presentations of CS + /US. The results demonstrate that the UR showed a greater amplitude when an ISI of 8 s was used than with ISIs of 1 or 5 s. No significant differences were found between the UR amp. of the 1- and 5-s ISI groups. These findings are discussed as a result of a hypothetical experimentally induced blend of the anticipatory CRs with the UR. PMID- 10461806 TI - ERP effects of spatial attention and display search with unilateral and bilateral stimulus displays. AB - Two experiments were performed in which the effects of selective spatial attention on the ERPs elicited by unilateral and bilateral stimulus arrays were compared. In Experiment 1, subjects received a series of grating patterns. In the unilateral condition these gratings were presented one at a time, randomly to the right or left of fixation. In the bilateral condition, gratings were presented in pairs, one to each side of fixation. In the unilateral condition standard ERP effects of visual spatial attention were observed. However, in the bilateral condition we failed to observe an attention related posterior contralateral positivity (overlapping the P1 and N1 components, latency interval about 100-250 ms), as reported in several previous studies. In Experiment 2, we investigated whether attention related ERP lateralizations are affected by the task requirement to search among multiple objects in the visual field. We employed a task paradigm identical to that used by Luck et al. (Luck, S.J., Heinze, H.J., Mangun, G.R., Hillyard, S.A., 1990. Visual event-related potentials index focused attention within bilateral stimulus arrays. II. Functional dissociation of P1 and N1 components. Electroencephalogr. Clin. Neurophysiol. 75, 528-542). Four letters were presented to a visual hemifield, simultaneously to both the attended and unattended hemifields in the bilateral conditions, and to one hemifield only in the unilateral conditions. In a focused attention condition, subjects searched for a target letter at a fixed position, whereas they searched for the target letter among all four letters in the divided attention condition (as in the experiment of Luck et al., 1990). In the bilateral focused attention condition, only the contralateral P1 was enhanced. In the bilateral divided attention condition a prolonged posterior positivity was observed over the hemisphere contralateral to the attended hemifield, comparable to the results of Luck et al. (1990). A comparison of the ERPs elicited in the focused and divided attention conditions revealed a prolonged 'search related negativity'. We discuss possible interactions between this negativity and attention related lateralizations. The display search negativity consisted of two phases, one phase comprised a midline occipital negativity, developing first over the ipsilateral scalp, while the second phase involved two symmetrical occipitotemporal negativities, strongly resembling the N1 in their topography. The display search effect could be modelled with a dipole in a medial occipital (possibly striate) region and two symmetrical dipoles in occipitotemporal brain areas. We hypothesize that this effect reflects a process of rechecking the decaying information of iconic memory in the occipitotemporal object recognition pathway. PMID- 10461807 TI - A startle-probe methodology for investigating the effects of active avoidance on negative emotional reactivity. AB - This study introduces a new methodology for investigating the impact of active avoidance and behavioral control on defensive emotional reactivity using the startle reflex. A between-groups yoked design was devised that permitted manipulation of participants' perception of control over an aversive event (loud noise) while precisely controlling motor activity and noxious stimulation. Startle responses to tactile (airpuff) probes were compared during threat/performance trials and neutral trials. Results conclusively demonstrated cross-modal startle potentiation in the context of a continuous motor performance task. Also, consistent with prior research, heart rate increased with perceived control. However, behavioral control per se did not appear to mitigate defensive emotion as indexed by startle potentiation. These findings indicate that other parameters may mediate the efficacy of active coping in addition to control, and that the startle probe paradigm can provide a valuable tool for investigating these parameters in future research. PMID- 10461808 TI - Molecular mechanisms that control leukocyte extravasation: the selectins and the chemokines. AB - Attachment of leukocytes to the blood vessel wall initiates leukocyte extravasation. This enables leukocytes to migrate to and accumulate at sites of tissue injury or infection where they execute host-defense mechanisms. A series of vascular cell adhesion molecules on leukocytes and on endothelial cells mediate leukocyte attachment to the endothelium in a stepwise process. A large panel of about 40 known human chemokines is able to specifically activate certain leukocytes and attract them to migrate across the endothelial barrier and within tissue. The specific combination of molecular signals provided by the diversity of cytokines, adhesion molecules, and chemokines regulates the specificity and selectivity of the recruitment of certain subpopulations of leukocytes in vivo. This review will focus on selectins and chemokines which initiate the cell contact and regulate activation and chemoattraction of leukocytes. PMID- 10461809 TI - Syndecan-4 expression is associated with follicular atresia in mouse ovary. AB - Ovarian granulosa cells synthesize heparan sulfate proteoglycans (HSPGs), that have anticoagulant properties. Moreover, HSPGs greatly increase in the granulosa cells during follicular atresia. However, the species of ovarian HSPGs have not yet been identified. Syndecan-4 (ryudocan, amphiglycan) is a membrane-spanning HSPG and a member of the syndecan family. Herein, we demonstrate that syndecan-4 is expressed in the granulosa cells of type 4-5b follicles and, most intensely, in those of the atretic follicles in the mouse ovary, as revealed by in situ hybridization. There is no relationship between syndecan-4 expression and age or sexual cycle stage. Compared with syndecan-4 expression, syndecan-1 and -3 are expressed more abundantly in postovulatory follicles and the corpora lutea, but less in the type 4-5b follicles and much less in the atretic follicles. Immunohistochemistry also demonstrates syndecan-4 expression in atretic follicles with apoptosis. The present study has revealed the distinct modes of expression of the syndecan family members, and the association of syndecan-4 expression and apoptosis in ovarian atretic follicles. PMID- 10461810 TI - Glucokinase is concentrated in insulin-secretory granules of pancreatic B-cells. AB - We immunohistochemically examined the distribution of glucokinase (GK) in the B cells of pancreatic islets of normal rats. GK was stained punctately in the cytoplasm of B-cells when examined under the light microscope. By use of a double immunostaining technique, most of the GK immunoreactivity was observed to be colocalized with insulin immunoreactivity. Electron microscopic examination by the immunogold method revealed that GK immunoreactivity was predominantly located within insulin-secretory granules of pancreatic B-cells. Exploration of the intracellular distribution of GK in hepatocytes suggested that the shuttling of the enzyme between the nucleus and the cytoplasm is essential for the regulation of GK activity (Toyoda et al. 1994, 1995, 1996a,b, 1997a). As an approach to the elucidation of the mechanism of control of GK activity, we immunohistochemically investigated the intracellular distribution of the enzyme in pancreatic B-cells under both light microscopy and electron microscopy in this study. A preliminary report of the present study has been published. PMID- 10461811 TI - Characterisation of caveolins from cartilage: expression of caveolin-1, -2 and -3 in chondrocytes and in alginate cell culture of the rat tibia. AB - This study was performed to determine if rat articular chondrocytes express caveolin, the structural protein of caveolae, and to determine differences in the distribution of the caveolin subtypes 1, 2 and 3 in knee joints of newborn and adult rats. All three subtypes of caveolin were detected in adult cartilage by immunocytochemical staining. In newborn rats, only caveolin-1 was found in the hyaline cartilage. Caveolin-1, -2 and -3 messenger RNA and protein were also detected in chondrocyte cell cultures. Ultrastructural investigations of cell culture and cartilage tissue revealed the presence of caveolae at the plasma membrane of chondrocytes. These findings represent the first report on the different expression of caveolin isoforms, in particular the expression of the muscle cell-specific caveolin-3 in chondrocytes. There is evidence that caveolin 2 and -3 are upregulated during growth and development of articular cartilage, suggesting a role for caveolins in chondrocyte differentiation. PMID- 10461812 TI - Fibronectin in human prostatic cells in vivo and in vitro: expression, distribution, and pathological significance. AB - In the present study we examined the expression and release of the extracellular matrix glycoprotein fibronectin (FN) in a prostate cancer cell line (LNCaP) and in primary prostatic stromal cells using the reverse transcription-polymerase chain reaction (RT-PCR) and by an enzyme-linked immunosorbent assay. Perturbation experiments in vitro using antibodies directed against FN and the FN receptor were also performed. Immunohistochemistry was used to show the in vivo distribution of FN and the FN receptor in tissue sections of normal human prostate, benign prostatic hyperplasia, and prostate carcinoma. The expression of the oncofetal FN ED-B segment in benign prostatic hyperplasia and prostate carcinoma tissue was investigated by RT-PCR. The FN mRNA was expressed by LNCaP and primary prostatic stromal cells, respectively. Both cell types released FN into the medium in a time-dependent manner, whereby FN secretion was about 2.5 fold higher in cultures of stromal cells relative to LNCaP cells. Blocking FN with anti-FN antibodies resulted in a significant decrease in cell adhesion for LNCaP cells and a change in morphology for the primary stromal cells. FN was located mainly in the stromal compartment of the prostate, showing a distinct distribution pattern in prostate carcinoma, whereas the FN receptor was detectable only in the prostate epithelia. RT-PCR experiments showed the expression of the oncofetal FN ED-B segment in benign prostatic hyperplasia and prostate carcinoma tissue, with a 3.5-fold higher expression in the prostate carcinoma probes. Our data point to an important role for FN in cell adhesion of prostatic cells and show that an alternatively spliced FN mRNA is upregulated in the pathologically altered human prostate. PMID- 10461813 TI - Increased spinal expression of c-Fos following stimulation of the lower urinary tract in chronic spinal cord-injured rats. AB - c-Fos expression was studied in the lumbar and sacral spinal cord regions involved in processing afferent input from the lower urinary tract and a comparison was made between spinal cord-injured (SCI) animals and control animals with intact neuraxes. Afferent pathways from the lower urinary tract were activated either by insertion of a catheter through the urethra into the urinary bladder or by catheterisation plus induction of reflex micturition contractions by intravesical saline infusion. Placement of a catheter alone elicited Fos expression in a similar number of neurones in SCI and control rats mainly in the medial dorsal horn (MDH) and dorsal commissure (DCM) in the segments L1-2 and L5 S1 with a maximum in L5. Additional saline infusion induced low-frequency, high amplitude, rhythmic bladder contractions of long duration in the rats with intact spinal cords, whereas in SCI rats, bladder distension elicited reflex contractions at a higher frequency, smaller amplitude and shorter duration. However, the basal and mean bladder pressure, as well as the total contraction time relative to the whole recording time, was not significantly different. Distension-induced bladder contractions markedly increased Fos expression primarily in the spinal segments L5-S1 in the control rats, where the majority of bladder and urethral afferent fibres terminates. Fos-positive cells were located in the MDH, lateral dorsal horn (LDH), DCM and the lateral aspect of laminae V VII. Compared to controls, Fos expression after spinal cord injury (SCI) occurred in a significantly greater number of neurones throughout the segments L3-S1 following induction of bladder reflexes. The greatest proportional increase in the number of Fos-positive cells occurred in L3-5 which normally receive only little afferent input from the urinary bladder. Cell numbers predominantly increased in the LDH and lateral lamina V-VII. The data are consistent with the concept of a neuroplastic reorganisation of spinal pathways after SCI. Unmasking of silent synapses or formation of new connections by afferent axonal sprouting caudal to the lesion, as evident from the increased numbers of cells expressing Fos after bladder distension, could be factors underlying the emergence of reflexogenic micturition in chronic SCI rats. PMID- 10461814 TI - Training affects myosin heavy chain phenotype in the trapezius muscle of women. AB - The aim of this investigation was to determine whether 10 weeks of three different types of training can alter the myosin heavy chain (MyHC) composition of the trapezius muscle. Twenty-one women were randomly assigned to three training groups that performed strength (n=9), endurance (n=7) or coordination training (n=5). Pre and post biopsies were taken from the upper part of the descending trapezius muscle and were analysed for MyHC isoform content using 5% gel electrophoresis. In addition, we have studied the expression of embryonic and neonatal MyHCs using double-immunofluorescence staining. In the strength-trained group, there was a significant increase in the amount of MyHC IIA and a significant decrease in the amount of MyHC IIB and MyHC I. In the endurance group, there was a significant decrease in the amount of MyHC IIB. MyHC composition in the coordination group was not altered. Following the training period, myotubes and individual small-sized muscle fibres were observed in the strength and endurance trained groups. These structures were stained with the markers for early myogenesis (MyHC embryonic and neonatal). These data suggest that specific shifts in MyHC isoforms occur in the trapezius muscle following strength and endurance training. The presence of small-sized muscle fibres expressing the developmental isoforms of MyHC suggests that strength and endurance training induced the formation of new muscle fibres. PMID- 10461815 TI - Localisation of actin, villin, fimbrin, ezrin and ankyrin in rat taste receptor cells. AB - Mammalian taste buds consist of 50-150 pear-or spindle-shaped taste receptor cells which contain, at their apical cell surface, a bundle of microvillar projections. The microvilli probably serve to increase the receptive membrane surface of the chemosensory receptor cells. The molecular basis controlling the ultrastructure of taste receptor microvilli is present unknown. In the present study we analysed, by immunostaining at the light and electron microscopic levels and by immunoblotting, components of the cytoskeleton of these microvilli. We show here that taste cell microvilli contain the major cytoskeletal proteins of intestinal microvilli, actin, fimbrin and villin. Another actin-binding, peripheral membrane protein of intestinal microvilli, ezrin, was also localised to taste cell microvilli, where ezrin might play a role, for example, in placement of specific membrane proteins to the microvillus membrane. In search of further linkage proteins, we found ankyrin localised along the basolateral cell surface of taste receptor cells, where ankyrin might be involved in the immobilisation of the Na+, K+-ATPase or other ion-translocating proteins of taste cells to the membrane cytoskeleton. PMID- 10461816 TI - Are there lessons from Littleton? PMID- 10461817 TI - Use of selective serotonin reuptake inhibitors for the treatment of childhood panic disorder: a pilot study. AB - This preliminary study examines the effectiveness and safety of selective serotonin reuptake inhibitors (SSRIs) for the treatment of panic disorder in children and adolescents. In a prospective open label study, 12 children and adolescents with panic disorder were treated with SSRIs, and if necessary, with benzodiazepines, for a period of 6-8 weeks and were followed for approximately 6 months. During the trial, clinician-based and self-report rating scales for anxiety and depression, functioning, and side effects, were administered. Using the Clinical Global Impression Scale (CGIS) 75% of patients showed much to very much improvement with SSRIs without experiencing significant side effects. After controlling for changes in depressive symptoms, self-report and clinician-based anxiety scales also showed significant improvement. At the end of the trial, 67% of patients no longer fulfilled criteria for panic disorder and 4 patients remained with significant residual symptoms. In conclusion, SSRIs appear to be a safe and promising for the treatment of children and adolescents with panic disorder, however, randomized controlled trials evaluating the effects of SSRIs and other interventions (e.g., cognitive therapy) for treating panic disorder in children and adolescents are warranted. It appears that until the SSRIs begin to exert their effects, a benzodiazepine adjunct treatment might be helpful for patients with severe panic disorder. PMID- 10461818 TI - Thermal response to serotonergic challenge and aggression in attention deficit hyperactivity disorder children. AB - Body temperature change in response to the serotonergic (5-HT) enhancer, d,l fenfluramine (FEN), was examined in 27 prepubescent boys diagnosed with Attention Deficit Hyperactivity Disorder (ADHD) to determine (1) the utility of this measure as an index of central serotonergic function; and (2) if the magnitude of temperature change is associated with aggression. FEN, 1 mg/kg, produced a significant increase in body temperature, the magnitude of which was correlated with plasma levels of the FEN metabolite, norfenfluramine (NORFEN). Furthermore, a significant inverse relationship was found between temperature response to FEN and teacher ratings of aggression. Parent ratings of aggression were not significantly correlated with the hyperthermic response to FEN. Interestingly, the magnitude of the hyperthermic response was unrelated to changes in plasma levels of prolactin and cortisol, suggesting that thermal and neuroendocrine responses are mediated by distinct 5-HT mechanisms. The agreement of these finding with those of studies using other procedures to assess the relationship between 5-HT and aggression suggests that decreased central 5-HT is associated with increased aggression. PMID- 10461819 TI - Risperidone-induced hepatotoxicity in children and adolescents? A chart review study. AB - Risperidone is an atypical antipsychotic drug that has been used in the treatment of numerous psychiatric disorders in children and adolescents. The question of whether risperidone-induced weight gain is associated with steatohepatitis has recently been raised. The purpose of this chart review was to ascertain: (1) the rate of liver dysfunction observed during risperidone treatment in children and adolescents; and (2) the clinical factors associated with liver dysfunction. For purposes of this chart review study, abnormal liver function was defined by serum transaminase or bilirubin values falling outside the normal laboratory ranges. Chart reviews were completed on 38 youths with ages ranging from 5-17 years with a variety of psychiatric diagnoses. The mean length of risperidone treatment was 15.2 months at a mean dose of 2.5 mg/day. It was found that 37 of the 38 youths treated with risperidone had no liver enzyme abnormalities at the end of study. One subject had an alanine aminotransferase (ALT) level of 46 U/L which was 7 U/L above the upper limit of normal for this laboratory test. This isolated value was not considered clinically significant. These data were noted in spite of weight gain and the use of numerous concomitant psychotropic medications. These findings suggest that risperidone in short term treatment does not commonly lead to evidence of abnormal liver function at therapeutic doses in children and adolescents. Larger-scale, prospective studies are needed in order to confirm these findings. PMID- 10461820 TI - Open-label quetiapine in the treatment of children and adolescents with autistic disorder. AB - OBJECTIVE: The purpose of this trial was to investigate the short-term safety and efficacy of quetiapine fumarate in the treatment of children and adolescents with autistic disorder (AD). METHOD: This was a 16-week, open-label trial that included 6 male subjects with a mean age of 10.9 +/- 3.3 years. All subjects met the DSM-IV criteria for AD and functioned in the mentally retarded range (mild, n = 2; moderate, n = 3; severe, n = 1). Behavioral ratings were obtained at baseline and every four weeks thereafter. RESULTS: Overall, there was no statistically significant improvement between baseline and endpoint for the group as a whole. Only two subjects completed 16 weeks of treatment and were considered "responders" by the global improvement item of the Clinical Global Impression Scale (CGIS). Dosages ranged from 100 to 350 mg/day (1.6-5.2 mg/kg/day). Subjects dropped out prematurely because of lack of response and sedation, limiting further dose increases (n = 3), and because of a possible seizure during the fourth week of treatment (n = 1). Other significant side effects included behavioral activation, increased appetite and weight gain (range, 0.9 to 8.2 kg). CONCLUSIONS: Quetiapine was poorly tolerated and associated with serious side effects in this clinical population. PMID- 10461821 TI - Phenelzine treatment of selective mutism in four prepubertal children. AB - Four children, ages 5 1/2 to 7 years, diagnosed with selective mutism and successfully treated with phenelzine are presented. All four children were anxious and extremely shy and had family histories of anxiety and shyness. The first three children were treated with phenelzine prior to reports that fluoxetine was helpful in some cases. The fourth child was treated initially with fluoxetine and had shown only minimal improvement after 10 months of doses up to 16 mg daily. Phenelzine doses ranged from 30 to 60 mg/day and treatment, including medication taper, lasted from 24 to 60 weeks. The phenelzine was generally well-tolerated and weight gain was the most common side effect. No hypertensive reactions or serotonin syndromes occurred. There was no recurrence of mutism after medication discontinuation. The authors conclude that phenelzine can be an effective treatment for selective mutism, but because of the possibility of serious food and drug interactions and the necessary dietary restrictions, it should be reserved for cases that do not respond to behavior therapy and fluoxetine or other specific serotonin reuptake inhibitors. PMID- 10461823 TI - Atrioventricular nodal re-entrant tachycardia associated with stimulant treatment. AB - A 13-year-old African-American female taking sertraline for obsessive compulsive disorder was diagnosed with her first episode of atrioventricular (AV) nodal re entrant tachycardia five days after beginning Mixed Salts of a Single-Entity Amphetamine Product (Adderall) for treatment of attention-deficit hyperactivity disorder (ADHD). She received successful cardioversion with 6 mg of intravenous adenosine, but developed a second episode of possible AV nodal re-entrant tachycardia twelve days after Adderall was reinitiated at half the previous dose. The patient had clinically similar cardiac episodes five and six months after treatment was changed to slow-release methylphenidate. Stimulant medication may evoke onset of AV nodal tachyarrhythmias in patients who have the potential to develop them, possibly in combination with a selective serotonergic reuptake inhibitor (SSRI). PMID- 10461822 TI - Risperidone augmentation of serotonin reuptake inhibitor treatment of pediatric obsessive compulsive disorder. AB - In this case series, risperidone augmentation of treatment with a serotonin reuptake inhibitor (SRI) is described in four pediatric patients diagnosed with obsessive compulsive disorder (OCD). An improved treatment response was observed in all cases, albeit in different ways. All four of the patients had failed prior SRI monotherapy. Comorbid tics were observed in two cases and aggressive behavior or violent images were seen in three. Possible predictors of response to risperidone in patients with OCD and future research avenues are explored. PMID- 10461824 TI - Pharmacokinetics and hemodynamic effects of single oral doses of thalidomide in asymptomatic human immunodeficiency virus-infected subjects. AB - Thalidomide (alpha-N-phthalimidoglutarimide), a potent inhibitor of tumor necrosis factor alpha (TNF-alpha), is proving to be a promising drug in the treatment of a number of inflammatory, autoimmune, and HIV-associated disorders. The pharmacokinetics and hemodynamic effects of two single oral doses of thalidomide (100 and 200 mg) were investigated, using a randomized, two-period crossover design, in a group of asymptomatic, male HIV-seropositive subjects. Thalidomide pharmacokinetics were linear at the doses studied, and were best described by a one-compartment model with first-order absorption and elimination processes. The drug was rapidly absorbed, with a mean absorption half-life of 0.95 hr (range, 0.16-2.49 hr) and 1.19 hr (range, 0.33-3.53 hr) after 100- and 200-mg doses, respectively. The corresponding mean Cmax values were 1.15+/-0.24 microg/ml (100 mg) and 1.92+/-0.47 microg/ml (200 mg; p<0.001), which were achieved (Tmax) at 2.5+/-1.5 h and 3.3+/-1.4 hr, respectively. Plasma concentrations of thalidomide declined thereafter, in a log-linear manner, with elimination half-lives of 4.6+/-1.2 hr (100 mg) and 5.3+/-2.2 hr (200 mg). The apparent volumes of distribution (Vdss/F) were 69.9+/-15.6 liters (100 mg) and 82.7+/-34.9 liters (200 mg) while total body clearances (Cl/F) were 10.4+/-2.1 and 10.8+/-1.7 liters/hr, respectively. Significant dose-dependent decreases in supine systolic and diastolic blood pressures were seen for up to 2 hr postdosing; somnolence, headache, dizziness, and confusion were also reported more frequently at the higher dose of thalidomide. PMID- 10461825 TI - Normal telomere lengths in naive and memory CD4+ T cells in HIV type 1 infection: a mathematical interpretation. AB - To study CD4+ T cell productivity during HIV-1 infection, CD4+ T cell telomere lengths were measured. Cross-sectional and longitudinal analysis of HIV-1 infected individuals with CD4+ T cells counts >300 cells/mm3 showed normal average telomeric restriction fragment (TRF) length and normal shortening rates of CD45RA+ naive and CD45RO+ memory CD4+ T cells. These TRF data were interpreted in terms of CD4+ T cell production by means of a mathematical model. This model resolves previous criticisms arguing that the normal TRF length of CD4+ T cells in HIV-1 clinical latency is due to the killing of dividing CD4+ T cells by the virus. Only an increased priming rate of naive CD4+ T cells to become memory cells may elongate the average TRF length of memory CD4+ T cells, and may therefore mask the shortening effect of increased turnover in the CD4+ memory T cell compartment. The data are more compatible with the notion that during HIV-1 clinical latency the turnover of CD4+ T cells is not markedly increased, however, and that HIV-related interference with renewal from progenitors plays a role in CD4+ T cell depletion. In such a "limited renewal" scenario disease progression is no longer a consequence of markedly increased CD4+ T cell production. PMID- 10461826 TI - Correlation between circulating stromal cell-derived factor 1 levels and CD4+ cell count in human immunodeficiency virus type 1-infected individuals. AB - Stromal cell-derived factor 1 (SDF-1) is the natural ligand that recognizes CXCR4, which also serves as a coreceptor for some strains of HIV-1. In this study, we explored SDF-1 blood levels among HIV-1-infected individuals exhibiting a wide range of CD4+ cell counts. Plasma or serum concentrations of SDF-1 protein were measured by ELISA in samples from 31 HIV-1-seronegative individuals and 79 HIV-1-infected subjects. Although SDF-1 protein levels were stable for months among seronegative individuals (mean intrasubject variation, 17%), the absolute values varied widely (0.28 to 106.5 ng/ml; mean, 25.6 ng/ml). In HIV-1-infected subjects, there was a direct correlation between SDF-1 level and CD4+ cell count. Subjects with fewer than 50 CD4+ cells per cubic microliter of blood had significantly lower mean SDF-1 levels (+/-SD) than did either HIV-1-infected subjects with higher CD4+ cell counts or uninfected controls: CD4+ cell count <50, mean SDF-1 level of 10.7+/-33.7, 50 < CD4+ cell count <200, mean SDF-1 level of 12.9+/-19.0, 200 < CD4+ cell count <500, mean SDF-1 level of 19.3+/-36.8; CD4+ cell count >500, mean SDF-1 level of 18.5+/-25.2; uninfected control mean SDF-1 level, 25.6+/-34.7. No significant change in SDF-1 level was detected after administration of antiretroviral therapy in nine subjects with advanced disease (mean intrasubject variation, 43%). Analysis of SDF-1 mRNA expression in lymph nodes from HIV-1-infected subjects at different disease stages revealed that the medullary cords contained stromal cells that express SDF-1 mRNA. This preliminary analysis suggests a possible link between lower SDF-1 levels and disease progression. PMID- 10461827 TI - Sustained suppression of plasma HIV RNA is associated with an increase in the production of mitogen-induced MIP-1alpha and MIP-1beta. AB - The chemokine receptor CCR5 has been shown to be a major coreceptor for HIV-1. The chemokines that bind to this receptor (MIP-1alpha, MIP-1beta, and RANTES) are potent inhibitors of HIV replication and may play an important role in the pathophysiology of HIV disease. We investigated the effect of potent antiretroviral therapy (ritonavir and saquinavir) on the production of MIP 1alpha, MIP-1beta, and RANTES in 19 HIV-infected patients who had sustained decreases in plasma HIV RNA levels (<200 copies/ml). Chemokine concentrations were measured in serum, plasma, and PHA-stimulated PBMCs at baseline and 24 and 48 weeks after initiating therapy. MIP-1alpha, MIP-1beta, and RANTES levels in serum and plasma did not significantly change in the 48-week period. In contrast, MIP-1alpha and MIP-1beta secreted by PHA-stimulated PBMCs increased at 24 weeks, with this increase sustained at 48 weeks, whereas no significant change was observed in PHA-induced RANTES production. A significant positive correlation was found between the changes in PHA-induced chemokine production and baseline CD4+ T cell counts. These data demonstrate that sustained suppression of viral replication by potent antiretroviral therapy has a potentially beneficial effect on chemokine production and early initiation of this therapy appears to confer a more favorable chemokine profile. PMID- 10461828 TI - Anti-CD4 antibodies in exposed seronegative adults and in newborns of HIV type 1 seropositive mothers: a follow-up study. AB - In this work, an ELISA for the quantitative determination of IgG anti-CD4 autoantibodies was validated and utilized in the follow-up of two cohorts of HIV 1-exposed seronegative subjects. A serum with an arbitrarily assigned concentration of 100,000 units/ml was used as a reference, and the detection limit, inter- and intraassay variability, and analytical recovery were calculated. The study subjects included adults sexually exposed to HIV-1-infected partners and the newborns of HIV-1+ mothers who seroreverted by 18 months of age. Some of these individuals were studied over an 18- to 24-month period. The detection limit of the assay was 2000 AU/ml. Intra- and interassay variability was, respectively, 3.92 and 3.90%. Analytical recovery in an assay in which a fixed amount of anti-CD4 antibodies was added to different samples was 98%. A proportion of adults (16 of 47, 34.0%) and babies (12 of 27, 44.4%) had significantly higher concentrations of anti-CD4 antibodies. Among them, 8 adults maintained the same concentration as that found in the first determination; on the other hand, 12 babies born to seronegative mothers showed a significant increase in the concentration of anti-CD4 antibodies during their first months of life. In conclusion, anti-CD4 antibodies can be measured using a validated ELISA. They represent a serologic trait that is quantitatively conserved in HIV-1 exposed seronegative adult individuals and is actively acquired by newborns to HIV+ mothers. PMID- 10461829 TI - Monoclonal antibodies recognize at least five epitopes on the SIV Nef protein and identify an in vitro-induced mutation. AB - Eleven monoclonal antibodies (MAbs) to SIV Nef were produced and characterized. Five antibody-binding sites on SIV Nef were identified on the basis of the reactivity of the antibodies with recombinant proteins. Two of the five epitopes were defined using overlapping peptides. A further three epitopes could not be defined with peptides but all antibodies reacted in Western blot, suggesting that the epitopes were at least partially conformation dependent. Antibodies in two of the five epitope groups were further differentiated by competition analysis. The panel of MAbs described is able to distinguish between a number of recombinant Nef proteins currently under investigation in vivo in macaques. Two of the MAbs described are able to distinguish between the Nef protein from pathogenic (J5) and attenuated (C8) strains of SIV, thus providing useful tools for studying the relevance of the Nef protein in the pathogenesis of SIV infection. In FACScan analysis two of the MAbs, KK70 and KK75, were used to identify an in vitro induced mutation in J5 Nef grown in C8166 cells. Sequence analysis of the phenotypic variants identified a mutation of the tryptophan (TGG) at amino acid 214 to a stop codon (TGA), thus truncating the Nef protein. The functional significance of this observation remains unclear but highlights the need to interpret data with caution if virus has been cultured in vitro even for a short period of time. PMID- 10461830 TI - Accumulation of specific amino acid substitutions in HLA-B35-restricted human immunodeficiency virus type 1 cytotoxic T lymphocyte epitopes. AB - HLA is one of the genetic factors that influence the clinical course of HIV-1 infection, and patients with HLA-B35 are prone to rapid disease progression. Nine viral epitopes that are recognized by cytotoxic T lymphocytes (CTLs) in an HLA B35-restricted manner were determined. To examine how HIV-1 sequences are selected by CTLs in vivo, we sequenced the nine CTL epitopes of the virus in patient plasma. Here we show that certain amino acid substitutions at three epitopes were observed with significantly higher frequency in HLA-B35-positive patients than in HLA-B35-negative patients. By performing experiments with CTL clones established from the HLA-B35-positive patients, it was determined that one of the three substitutions was probably an escape mutation. However, concerning the other two epitopes, representative CTL clones killed target cells pulsed with mutant peptides as efficiently as those pulsed with wild-type peptides, suggesting that CTLs that can be established in vitro are not functioning properly in vivo. Amino acid sequence drift in all HLA-B35-restricted epitopes was rare during the observation period (1 year). Our results may have relevance in understanding the rapid clinical progression in HLA-B35-positive patients. PMID- 10461831 TI - Sendai virus-based production of HIV type 1 subtype B and subtype E envelope glycoprotein 120 antigens and their use for highly sensitive detection of subtype specific serum antibodies. AB - We previously described a Sendai virus (SeV)-based expression system for the recombinant gp120 of HIV-1 subtype B (rgp120-B), which has permitted the production of antigenetically and functionally authentic gp120 at a concentration as high as 6 microg/ml of culture supernatant (Yu D et al.: Genes Cells 1997;2:457-466). Here the same procedure was successfully applied to the production of HIV-1 subtype E gp120 (rgp120-E). The remarkable production of the proteins by the SeV expression system enabled us to use crude culture supernatants for serological and functional studies of gp120s. The immunological authenticity of rgp120-E was verified by patient sera and anti-V3 loop monoclonal antibodies specific for HIV-1 subtypes B and E. CD4-binding properties were corroborated by FACS analyses. The rgp120s were then used in an enzyme immunoassay (rgp120-EIA) to detect antibodies in the sera of HIV-1-infected individuals, and the performance was assessed in comparison with a conventional V3 loop peptide EIA (V3-EIA). The initial evaluation of a serum panel (n = 164) consisting of 76 subtype E and 88 subtype B sera revealed that the rgp120-EIA was nearly 1000-fold more sensitive than the V3-EIA and was able to detect subtype specific antibody with 100% sensitivity and with a complete correlation with the genotypes, whereas the V3-EIA failed to detect 9 and 24% of the same subtype E and B sera, respectively. Furthermore, a study employing a panel of 28 international sera with known genotypes (HIV-1 subtypes A through F) confirmed the remarkable specificity of this method. An EIA reactivity higher than 1.0 was an unambiguous predictor of HIV-1 subtype E and B infections. The data imply the presence of strong subtype-specific epitopes for antibody bindings to these rgp120s. PMID- 10461832 TI - Combined systemic and mucosal immunization with microsphere-encapsulated inactivated simian immunodeficiency virus elicits serum, vaginal, and tracheal antibody responses in female rhesus macaques. AB - We determined the efficacy of immunization with microsphere-encapsulated whole inactivated simian immunodeficiency virus (SIV) by combined systemic and mucosal administration to protect female rhesus macaques against vaginal challenge with homologous rhesus PBMC-grown SIVmac251. Animals in one group were primed and boosted intramuscularly. Two groups were primed intramuscularly and boosted either intratracheally or orally. A final group was primed by vaccinia/rgp140 scarification and subdivided for either intratracheal or oral boosting. Strong ELISA titers of circulating SIV-specific IgG and modest IgA responses were elicited in the animals primed intramuscularly. Intratracheal boosting in the intramuscularly primed macaques resulted in high bronchial alveolar wash (BAW) IgG and less pronounced IgA. SIV-specific vaginal wash (VW) IgG was also present in the intramuscular/intramuscular and intramuscular/intratracheal groups. Vaccinia/rgp140 priming gave low ELISA titers to whole SIV, and failed to elicit mucosal antibody regardless of the booster route. No animal in any group developed serum neutralizing antibody to homologous SIVmac251. On vaginal challenge none of the immunized groups was infected at a lesser frequency than the unimmunized controls. These data suggest that the use of microspheres in a combined parenteral and mucosal regimen is an effective method of eliciting IgG and IgA antibody at mucosal surfaces. PMID- 10461833 TI - Domestic transmission of HIV type 1 subtype G strains in Taiwan. PMID- 10461834 TI - Analysis of the V3 loop sequences from 12 HIV type 1-infected patients from Colombia, South America. PMID- 10461835 TI - Isolation of HIV type 1 from long-term nonprogressors in Herpesvirus saimiri immortalized T cells. PMID- 10461836 TI - The role of glycopeptide antibiotics in the treatment of infective endocarditis. AB - There are several sets of guidelines for the treatment of infective endocarditis, reflecting the need for differing treatment in various countries and times. This review considers the need for differing treatment modalities and in particular the utility of the glycopeptide antibiotics vancomycin and teicoplanin. Specific recommendations are offered as to when to consider the use of glycopeptides, appropriate dosage, length of treatment course and whether to use monotherapy or combined therapy. Used judiciously, the glycopeptides give results as good as can be achieved with other antimicrobial agents without exceptional toxicity. The potential of teicoplanin for use in the outpatient treatment of infective endocarditis is considered. PMID- 10461837 TI - Correlation between consumption of antimicrobials in humans and development of resistance in bacteria. AB - The correlation between consumption of antimicrobials in humans and the emergence of resistance in bacteria is complex and has proved difficult to establish. Besides antimicrobial use, many other distinct contributing factors are also involved in the issue. Despite this complexity, there is a substantial body of evidence that the use of antibiotics in prophylaxis and in therapy is associated with the development of resistance in the hospital and in the community. Some examples are reviewed, including increase of resistance in enterobacteriaceae, Pseudomonas aeruginosa, Acinetobacter spp., Streptococcus pneumoniae, Staphylococcus aureus, Coagulase Negative Staphylococci and Streptococcus pyogenes after the use of beta-lactam antibiotics, aminoglycosides, fluoroquinolones and macrolides. Success in reversing the rise of resistant strains has been rarely described. Two examples are highlighted, the reduction in the incidence of nasal carriage of penicillin-resistant pneumococci in Icelandic children, and a significant decline in erythromycin resistance in S. pyogenes after the reduction in the use of macrolides in Finland. PMID- 10461838 TI - Molecular epidemiology of Plasmodium falciparum antifolate resistance in Vietnam: genotyping for resistance variants of dihydropteroate synthase and dihydrofolate reductase. AB - Using PCR techniques, we analysed the dihydropteroate synthase (DHPS) mutations associated with sulphonamide resistance and the dihydrofolate reductase (DHFR) mutations associated with resistance to pyrimethamine and cycloguanil in samples from Plasmodium falciparum-infected Vietnamese patients. Of the 40 samples analysed, 39 had DHFR mutations associated with high level resistance to pyrimethamine, whereas only three had mutations at position 164, which is linked to cross resistance to both DHFR inhibitors. The DHPS, 437Gly variant associated with very mild resistance to sulphadoxine was found in 38 out of the 40 samples. Of seven samples resistant to Fansidar in vivo, only two were fully explained by the currently documented DHPS mutations. The treatment failure could be due to a high level of pyrimethamine resistance caused by the detected mutations. Most patients, however, were cured with a single dose of Fansidar in spite of the high number of resistance mutations found. PMID- 10461839 TI - Reduced susceptibility in laboratory-selected mutants of Aspergillus fumigatus to itraconazole due to decreased intracellular accumulation of the antifungal agent. AB - To study the mechanism of resistance of Aspergillus fumigatus to itraconazole, spontaneous mutants with reduced susceptibility were selected by spreading 2 x 10(8) conidia from a clinical isolate (W73355) susceptible to miconazole (MIC 2 mg/l), itraconazole (MIC 0.25 mg/l) and amphotericin B (MIC 0.5 mg/l) on 40 peptone yeast extract glucose agar plates containing miconazole (32 mg/l). The 19 colonies that grew (frequency 0.95 x 10(-7)) in the presence of miconazole were screened by broth macrodilution technique for their susceptibility to itraconazole. A total of two isolates (frequency 1 x 10(-8)) MCZ14 and MCZ15 had MICs of 16 and 8 mg/l, respectively, for itraconazole. Both MCZ14 and MCZ15 showed concomitant increases in MICs for ketoconazole and miconazole, but not for amphotericin B. Growth inhibition studies as well as kill curve experiments revealed that MCZ14 and MCZ15 were less susceptible to itraconazole compared to the parental strain. The intracellular accumulation of itraconazole in A. fumigatus was time and concentration dependent. Maximum accumulation was obtained within 30 min at 5 microM itraconazole. In MCZ14 and MCZ15 intracellular accumulation of [3H]itraconazole was reduced by approximately 80 and 60%, respectively, compared to the susceptible parent. The respiratory inhibitor carbonyl cyanide m-chlorophenyl hydrazone at 200 microM reduced the intracellular accumulation of itraconazole by approximately 36.2% (P < or = 0.05) in the parent and in the mutant strains. These results suggest that (i) the reduced accumulation of itraconazole in MCZ14 and MCZ15 is due to diminished permeability of the drug and perhaps not due to efflux, (ii) the uptake of itraconazole in A. fumigatus may be an energy dependent process, and (iii) decreased accumulation of itraconazole is at least in part responsible for the reduced susceptibility of the mutant isolates to itraconazole. PMID- 10461840 TI - Efficacy of NND-502, a novel imidazole antimycotic agent, in experimental models of Candida albicans and Aspergillus fumigatus infections. AB - In vitro and in vivo anti-Candida albicans and anti-Aspergillus fumigatus activities of NND-502, a new imidazole-antimycotic, were compared with those of fluconazole (FCZ), itraconazole (ITZ) and/or amphotericin B (AmB). NND-502 exhibited strong in vitro antifungal activity against both fungal species; its MIC against C. albicans was 1-4 times lower than that of FCZ, and its MIC against A. fumigatus was at least 60-2000 times lower than that of ITZ and AmB. In vivo antifungal treatments with each drug were initiated 1 h after inoculation in the experimental models, so that antifungal potential reflected prophylactic activity rather than therapeutic activity. The oral regimen of NND-502 in a murine model of systemic C. albicans infection was much less effective than that of FCZ. In vivo anti-A. fumigatus activity of oral NND-502 in a murine model of systemic infection was apparently superior to that of FCZ and ITZ in terms of prolonging survival. In addition to the murine model of systemic aspergillosis, intravenous NND-502 was shown to be highly effective in a rat model of pulmonary aspergillosis compared with intravenous AmB; 90% of animals survived at a dose of 2.5 mg/kg per day of NND-502 while only 30% of animals escaped death when 5 mg/kg per day of AmB was used. This potent efficacy of NND-502 was also confirmed in a sublethal challenge study in which the administration of the agent at a dose as low as 1.25 mg/kg per day resulted in the significant reduction of organisms in the lung; no comparable effect of AmB was found. PMID- 10461841 TI - Postantibiotic and physiological effects of tilmicosin, tylosin, and apramycin at subminimal and suprainhibitory concentrations on some swine and bovine respiratory tract pathogens. AB - The antimicrobial activity of tilmicosin, tylosin, and apramycin on some important gram-negative swine and bovine pathogens namely, Pasteurella multocida, Pasteurella haemolytica, Bordetella bronchiseptica, and Actinobacillus pleuropneumoniae were studied in vitro. The effect of minimal inhibitory concentrations (MICs) and sub-MICs (1/4, 1/2 MIC) on bacterial growth was evaluated. The presence of tilmicosin, tylosin and apramycin in the medium decreased the rate of growth of the bacterial strains tested using drug concentrations as low as 1/4 MIC. The postantibiotic effect (PAE) which is the suppression of optimal bacterial growth that persists after a short exposure (2 h) of microorganisms to an antibiotic was studied by exposure of bacteria to drugs at 1/4, 1/2, 1, 4 and 8 times MIC. The duration of PAEs increased with rising concentration for all drugs tested but at concentrations below the MIC, tilmicosin showed more significant PAEs than tylosin or apramycin against P. multocida and A. pleuropneumoniae. Tilmicosin and tylosin caused PAEs of up to 8 h when used at 8 times the MIC, while apramycin caused PAEs of up to 5 h when used at this concentration. Sub-MICs of either tilmicosin, tylosin, or apramycin had no effect on P. multocida dermonecrotic toxin production. However sub-MICs of tylosin, or apramycin significantly reduced the haemolytic activity of A. pleuropneumoniae and affected the capsular material production of this isolate and of one isolate of P. multocida (type A). The in vitro effect of tilmicosin, tylosin, and apramycin (even when used at sub-MIC levels) on growth, production of capsular material, and haemolytic activity might impair the virulence of some of the microorganisms studied. In addition to the effects of these drugs on some putative virulence factors, we suggest that the strong PAEs caused by tilmicosin, tylosin, and apramycin may also contribute to the in vivo efficacy of these drugs. PMID- 10461842 TI - Enhanced susceptibility of pentylenetetrazole kindled mice to quinolone effects. AB - The present study was designed to examine the ability of different quinolones to affect the seizure severity and the latency of development of chemical kindling produced by repeated treatment using a subconvulsant dose of pentylenetetrazole (PTZ). A group of mice (kindled control) were treated subcutaneously (s.c.) with vehicle + PTZ (30 mg/kg, three times a week) for 6 consecutive weeks and the changes in excitability associated with the kindling state were observed over the following 2 h. A second group of mice were injected intraperitoneally (i.p.) with the following quinolone derivatives, ciprofloxacin (ciprox), pefloxacin (peflox), ofloxacin (oflox), cinoxacin (cinox), nalidixic acid (nalidixic), 1-cyclopropyl-6 amino-7-tetrahydroisoquinoline-8-methyl-4-oxo-1,4-dihydr oquinoline-3-carboxylic acid (M5) and 1-cyclopropyl-7-tetrahydro-isoquinoline-8-methyl-4-oxo-1,4 dihydroquinol ine-3-carboxylic acid (MH5) at a dose of 20 mg/kg 15 min before receiving a subconvulsant dose of PTZ (30 mg/kg, s.c.). The results showed that pretreatment with some of the quinolones tested facilitated the development of kindling to PTZ-induced seizures. In particular, ciprox, peflox, oflox, M5 and MH5 derivatives variously increased the development of kindling to PTZ induced seizures, whilst cinox and nalidix did not significantly affect it. Additionally we determined whether the enhanced susceptibility of kindled mice only occurred after relatively short intervals following the last seizure or whether it was a more permanent phenomenon. For the study of the persistence of kindling, the animals were rechallenged with the kindling stimulus (PTZ 25 mg/kg, s.c.) 15 and 30 days after the last injection of the chronic treatment with PTZ (30 mg/kg, s.c.) and the behavioural changes in the kindled mice were compared with the control ones (chronically treated with vehicle). The present data demonstrated that kindling produced long-lasting alterations, substantiating that epileptogenesis initiated by kindling renders the brain more susceptible to central nervous system (CNS) side effects of quinolones. An interaction between PTZ and quinolone derivatives which involves either an inhibition of gamma aminobutyric acid (GABA) neurotransmission or/and an increase in the function of the excitatory amino acid (EAA) system is suggested. PMID- 10461843 TI - Vaginal disinfection with chlorhexidine during childbirth. AB - The purpose of this study was to determine whether chlorhexidine vaginal douching, applied by a squeeze bottle intra partum, reduced mother-to-child transmission of vaginal microorganisms including Streptococcus agalactiae (streptococcus serogroup B = GBS) and hence infectious morbidity in both mother and child. A prospective controlled study was conducted on pairs of mothers and their offspring. During the first 4 months (reference phase), the vaginal flora of women in labour was recorded and the newborns monitored. During the next 5 months (intervention phase), a trial of randomized, blinded placebo controlled douching with either 0.2% chlorhexidine or sterile saline was performed on 1130 women in vaginal labour. During childbirth, bacteria were isolated from 78% of the women. Vertical transmission of microbes occurred in 43% of the reference deliveries. In the double blind study, vaginal douching with chlorhexidine significantly reduced the vertical transmission rate from 35% (saline) to 18% (chlorhexidine), (P < 0.000 1, 95% confidence interval 0.12-0.22). The lower rate of bacteria isolated from the latter group was accompanied by a significantly reduced early infectious morbidity in the neonates (P < 0.05, 95% confidence interval 0.00-0.06). This finding was particularly pronounced in Str. agalactiae infections (P < 0.0 1). In the early postpartum period, fever in the mothers was significantly lower in the patients offered vaginal disinfection, a reduction from 7.2% in those douched using saline compared with 3.3% in those disinfected using chlorhexidine (P < 0.05, 95% confidence interval 0.01-0.06). A parallel lower occurrence of urinary tract infections was also observed, 6.2% in the saline group as compared with 3.4% in the chlorhexidine group (P < 0.01, 95% confidence p interval 0.00-0.05). This prospective controlled trial demonstrated that vaginal douching with 0.2% chlorhexidine during labour can significantly reduce both maternal and early neonatal infectious morbidity. The squeeze bottle procedure was simple, quick, and well tolerated. The beneficial effect may be ascribed both to mechanical cleansing by liquid flow and to the disinfective action of chlorhexidine. PMID- 10461844 TI - Clinical and microbiological efficacy of single dose cefuroxime prophylaxis for dental surgical procedures. AB - In a prospective controlled randomised trial of 59 patients the effect of a single dose of cefuroxime before multiple tooth extractions on the clinical findings and occurrence of bacteraemia was studied. In one group 1.5 g cefuroxime was administered intravenously 10 min before start of surgery. A total of 118 blood samples were collected after 10 and 40 min. The cefuroxime group had a significantly lower rate of bacteraemia (33%) compared with the control group (86%). The predominant bacteria were gram-positive cocci. Fifty four different strains of bacteria were isolated, of which most were susceptible to cefuroxime. However seven strains which were susceptible to cefuroxime in vitro, were isolated from blood cultures of the treated group. Serum concentrations of cefuroxime ranged from 52.3 to 141.6 mg/l at 10 min and 32.6 to 91.8 mg/l at 40 min with a mean of 78.6 and 55.1 mg/l respectively. The only other factor which had a significant influence on the rate of bacteraemia was the number of extracted teeth. Oral hygiene and periodontal status did not significantly affect bacteraemia. There were no wound infections in any of the groups. During these dental interventions, sufficient plasma levels of antibiotic prevented bacteraemia. Selected patients might benefit from single-dose-prophylaxis with cefuroxime in preparation for dental surgery. PMID- 10461845 TI - Antimicrobial activity of spices. AB - Spices have been shown to possess medicinal value, in particular, antimicrobial activity. This study compares the sensitivity of some human pathogenic bacteria and yeasts to various spice extracts and commonly employed chemotherapeutic substances. Of the different spices tested only garlic and clove were found to possess antimicrobial activity. The bactericidal effect of garlic extract was apparent within 1 h of incubation and 93% killing of Staphylococcus epidermidis and Salmonella typhi was achieved within 3 h. Yeasts were totally killed in 1 h by garlic extract but in 5 h with clove. Some bacteria showing resistance to certain antibiotics were sensitive to extracts of both garlic and clove. Greater anti-candidal activity was shown by garlic than by nystatin. Spices might have a great potential to be used as antimicrobial agents. PMID- 10461846 TI - Imipenem for the treatment of melioidosis. AB - Melioidosis is a protean disease caused by Burkholderia pseudomallei. It is rare in the UK and is generally only seen in patients with a travel history to endemic areas such as Thailand, Singapore and Malaysia. Cases may present with disseminated bacteraemic, non-disseminated bacteraemic, multi-focal bacteraemic or localized disease. Subclinical infections also occur. Following acquisition of the organism a patient may remain asymptomatic for several years before infection becomes clinically apparent. Factors such as diabetes, renal failure or other causes for a decrease in host immunity may precipitate the appearance of overt disease. The current treatment choice for severe melioidosis is parenteral ceftazidime followed by oral amoxycillin-clavulanic acid or a combination of co trimoxazole, doxycycline and chloramphenicol. We report a case of melioidosis in a 59-year-old male diabetic from Bangladesh who initially responded to piperacillin-tazobactam but was changed to ceftazidime when a definitive diagnosis was made. His condition deteriorated on the latter antibiotic. He subsequently responded to imipenem. The patient's long-term outcome is still not known. PMID- 10461847 TI - Activity of pradimicin BMS-181184 against Aspergillus spp. AB - The pradimicins are a new class of antifungal agents with activity against the majority of human fungal pathogens. In this study, the in vitro activity of pradimicin BMS-181184 was investigated against a range of the most common species of Aspergillus. The results were compared with itraconazole and amphotericin B. BMS-181184 was found to be active against most Aspergillus spp., but at higher concentrations than itraconazole and amphotericin B. PMID- 10461848 TI - Estramustine resistance. AB - Estramustine (EM), a conjugate of nornitrogen mustard and estradiol, is a antimicrotubule drug currently in use for the treatment of advanced prostatic carcinoma. Experimental data are accumulating concerning the antitumor effect of EM in other malignancies, and clinical studies in other malignancies are ongoing. This review summarizes the information available to date concerning the effects of EM and the development of drug resistance. EM depolymerizes microtubules by binding to microtubule-associated proteins (MAPs) as well as tubulin. Because of the radiosensitizing effect of this drug there has been a recent increase in interest concerning estramustine and its clinical use. Recently, it was proposed that EM induces an apoptotic cell death in glioma cells in vitro and in a rat model. EM resistance is distinct from MDR phenotype; it has been used in combination with antimitotic agents which are part of the MDR phenotype. Observations made with estramustine-resistant cell lines show the acquisition of estramustine resistance is a function of multiple adaptation by changes at tubulin expression pattern, and is also associated with changes in tau expression and phosphorylation. PMID- 10461849 TI - Effects of nicardipine and bupivacaine on early after depolarization in rabbit sinoatrial node cells: a possible mechanism of bupivacaine-induced arrhythmias. AB - The effects of nicardipine and bupivacaine on early afterdepolarizations (EADs) were investigated in rabbit sinoatrial (SA) nodes using the conventional microelectrode technique. In a nominally Ca2+ -free, Mg2+ -free solution, addition of 0.5 mM Sr2+ produced EADs following prolongation of action potential duration. Nicardipine (10 microM) as well as Mg2+ (1 mM), both of which block the L-type Ca2+ channel current (iCa,L), abolished Sr2+ -induced EADs. Bupivacaine (5 microM), blocking the delayed rectifier K+ current (iK), facilitated the generation of EADs in the Sr2+ solution containing 1 mM Mg2+. The EADs in Sr2+ solution and the effect of bupivacaine were well simulated by the mathematical model when enhancement of slowly inactivating iCa,L and suppression of iK were assumed. Bupivacaine may cause sinus arrhythmias by facilitating EAD generation in SA node cells. PMID- 10461850 TI - Vasodilating effect of benidipine hydrochloride in the renal and hindquarter vascular regions (supplied by terminal aorta) of spontaneously hypertensive rats. AB - One of two Ca antagonists, benidipine (3-30 microg/kg) or nifedipine (30-600 g/kg), was administered in a bolus injection through the jugular vein, and the changes in mean arterial pressure (MAP), renal flow (RF), and hindquarter flow (HQF) in conscious spontaneously hypertensive rats (SHRs) and normotensive control rats (NCRs). Renal vascular resistance (RR) and hindquarter resistance (HQR) were calculated as MAP divided by RF and HQF, respectively. When a high dose was administered to decrease the blood pressure by about 20%, the RR was significantly lower with benidipine than with nifedipine. The decrease in HQR was not significantly different between benidipine and nifedipine. When a low dose was administered to decrease the blood pressure by about 7%, the decrease in RR was not significantly different between benidipine and nifedipine, but the HQR was significantly lower with benidipine than with nifedipine. In the NCRs, no pharmacological properties were significantly different between these two Ca antagonists. PMID- 10461851 TI - Taurine and calcium interaction in protection of myocardium exposed to ischemic reperfusion injury. AB - We aimed to investigate the cardio-protective role of taurine with low calcium level against reperfusion damage by adding taurine to extracellular fluid. Guinea pig hearts were mounted on Langendorf perfusion apparatus and different compositions of perfusion solutions were prepared for each experimental group. After 20 min of normothermic ischemia the hearts were reperfused. Pre-ischemic, post-ischemic and post-reperfusion percentage changes of heart rate and contractile force were compared. Post-reperfusion tissue weight, malondialdehyde (MDA) and prostaglandin E-like activity (PGE-like activity) were assessed. Taurine-added low-calcium perfusion solution significantly decreased the postischemic myocardial injury. PMID- 10461852 TI - Effects of adenosine receptor agonists on induction of contractions to phenylephrine of guinea-pig aorta mediated via intra- or extracellular calcium. AB - The vasorelaxant actions of adenosine and its analogue, 5'-(N-ethylcarboxamido) adenosine (NECA), were investigated in guinea-pig isolated aortic rings by addition to the tissue prior to induction of a contraction by the alpha1 adrenoceptor agonist phenylephrine (PE, 3x10(-6) M). The effect was calculated from the ratio (C2/C1) of the contraction to PE before (C1) and in the presence of adenosine or NECA (C2). This was compared with a control ratio obtained at the same time in which no vasorelaxant was present during C2. Experiments were performed in either "normal" or "Ca2+ -free" bathing medium. Both adenosine and NECA caused inhibition of contractions in "normal" and "Ca2+ -free" conditions, the latter indicating that the vasorelaxant action was due in part to inhibition of intracellular Ca2+ mobilization. To determine whether inhibition of influx of extracellular Ca2+ is a target for the vasorelaxation, contractions to PE were obtained in "normal" Ca2+ and in the presence of ryanodine (10(-5) M), which prevents the release of intracellular Ca2+. These contractions were inhibited by NECA indicating that stimulation of A2-receptors by NECA interferes with the influx of Ca2+ via the opening of receptor-operated Ca2+ channels (ROCs). This study has demonstrated that cell surface A2-receptor stimulation in the guinea pig aorta inhibits phenylephrine-induced contractions by interfering with both the release of intracellular Ca2+ and the influx of extracellular Ca2+, presumably via ROCs. PMID- 10461853 TI - Pharmacological actions of monovalent ionophores on spontaneously beating rabbit sino-atrial nodal cells. AB - The effects of sodium (monensin) and potassium (nigericin and lonomycin A) ionophores on the spontaneous activity in rabbit sino-atrial (SA) nodal cells were investigated using microelectrode and whole-cell voltage-clamp techniques. In the multicellular preparations, the ionophores produced a negative chronotropic effect in a concentration-dependent manner, and at 3x10(-5) M significantly decreased the amplitude and duration of action potentials and enhanced the maximum rate of depolarization. The ionophores elicited dysrhythmias and then a sinus arrest often occurred. These responses were reversible. In whole cell clamp experiments, monensin enhanced the L-type Ca2+ current (ICa), whereas lonomycin A and nigericin inhibited ICa. The fast component of the inactivation phase for ICa was decreased by the ionophores, but the slow component was unaffected. The activation and inactivation kinetics (d infinity and f infinity) were not altered. The ionophores did not affect the hyperpolarization-activated inward current. Monensin inhibited the delayed rectifier K+ current (I(K)), but lonomycin A and nigericin increased I(K). Its activation kinetics shifted in the depolarizing direction. The effects on the ionic currents were irreversible. Monensin (30 microM) increased cellular Ca2+ concentration ([Ca2+]i), using Ca2+ sensitive fluorescent dye (fura-2). These results indicate that the monovalent ionophores depress the action potentials and produce a negative chronotropic effect due to direct and indirect modulations of the ionic currents and the [Ca2+]i level in rabbit SA nodal cells. PMID- 10461854 TI - YoshixolTR inhibits B16 melanoma cell growth in vivo and induces apoptosis-like (quantum thermodynamic) cell death. AB - In this report, antitumor effects of YoshixolTR in vivo and in vitro were investigated in B16 melanoma cells. For in vivo experiments, the present study shows a dramatic inhibition of tumor growth of B16 melanoma transplanted on the leg or intraperitoneal cavity after treatment with YoshixolTR intraperitoneally. A proliferation of B16 cells in vitro was inhibited by YoshixolTR in a dose-and time-dependent manner. YoshixolTR induced apoptosis-like cell death in histological observations (phase-contrast, scanning and transmission electron microscopy), DNA fragmentation, and a smaller increase in lactate dehydrogenase (LDH) as a marker of cell leakage. Immunohistochemical investigation of cytoskeletal components, such as actin and tubulin, showed a cell wall disruption of B16 melanoma cells and a nuclear extrusion after the treatment with YoshixolTR. Treatment with YoshixolTR in vitro showed an arrest at the G0/G1 stage of the cell cycle, followed by a flow cytometric measurement. As a possible physiological mechanism of YoshixolTR on B16 melanoma cells, intracellular Ca++ was measured with Fura-2 technique. An adequate concentration of YoshixolTR, which induces apoptosis-like cell death, showed a decrease in intracellular free Ca++ concentration. In conclusion, YoshixolTR has an antitumor potency with a new biological mechanism of cell growth, proliferation, and differentiation, including cellular signalling pathways, and is a new candidate for an ideal chemotherapeutic agent against malignant tumors. PMID- 10461855 TI - Effect of bupranolol for BRL37344 and noradrenaline-induced relaxations mediating atypical beta/beta3-adrenoceptor in rat oesophageal muscularis mucosae. AB - We previously suggested that the existence of atypical beta/beta3-adrenoceptor with pA2-values for bupranolol, a non-selective beta-adrenoceptor antagonist, against BRL37344 and noradrenaline were 5.79 and 5.53 in guinea pig taenia caecum, respectively. We furthermore determined the affinity of bupranolol to subclassify atypical beta/beta3-adrenoceptor in rat oesophageal muscularis mucosae, because it is rich in atypical beta/beta3-adrenoceptor. BRL37344 and noradrenaline produced a concentration-dependent relaxation of rat oesophageal muscularis mucosae. The responses to BRL37344 and noradrenaline were resistant to 3x10(-6) M propranolol, 10(-4) M atenolol, and 10(-4) M butoxamine. However, bupranolol antagonized the responses to BRL37344 and noradrenaline in a concentration-dependent manner. Schild plot analyses of bupranolol against BRL37344 and noradrenaline gave pA2-values of 7.06 and 6.96, respectively. These results suggest that bupranolol can distinguish the difference in affinity between atypical beta/beta3-adrenoceptors in rat oesophageal muscularis mucosae and guinea pig taenia caecum. The difference in behavior of bupranolol confirms the existence of some atypical beta/beta3-adrenoceptors subtypes. PMID- 10461856 TI - In vitro and in vivo antitumor activity of YoshixolTR against murine L1210 leukemic cells. AB - In this report, antiproliferative effects of YoshixolTR in vitro and in vivo were investigated in murine L1210 cells. A proliferation of L1210 cells in vitro was inhibited by YoshixolTR in a dose- and time-dependent manner. This inhibition showed an arrest at the G0/G1 stage of the cell cycle, followed by a flow cytometric measurement. YoshixolTR induced apoptosis-like cell death identified by histological observations (scanning electron and transmission electron microscopy), DNA fragmentation, and a smaller increase in lactate dehydrogenase (LDH). In the in vivo experiments, YoshixolTR (5 microl/kg of body weight, on days 1, 3, and 5) was injected intraperitoneally in mice inoculated with L1210 cells. No marked prolongation of survival occurred between the control group and treated group. However, a survival curve in the treated group showed a shift toward a possible longer survival time. Additionally, on the basis of apoptosis like cell death due to YoshixolTR as indicated above, a possibility of immunotherapy as a tumor vaccine has been examined. A vaccination of rabbit anti serum, which consisted of components from the L1210 cells killed by YoshixolTR, produced a dramatic improvement of viability in the leukemic mice. In conclusion, YoshixolTR has an anti-leukemic potency with a new biological mechanism and an inductive potency of super-antigens as immunotherapeutic agents against malignant tumors. PMID- 10461857 TI - Dual action of the benzodiazepine receptor inverse agonist RU34347 on responses to exogenously applied GABA in the rat cerebellar slice. AB - The benzodiazepine receptor inverse agonist has been shown to produce agonist like effects at low concentrations. RU34347 has both inverse agonist (attenuation of GABA-responses) and agonist-like (reduction of spontaneous Purkinje cell firing rate) in the cerebellar slice preparation. The benzodiazepine antagonist flumazenil prevented the inverse agonist actions, but only partially reduced the agonist-like effects. Further, brief application of RU34347 to slices mimicked the response to GABA, and pharmacological investigation determined that this action was mediated through increased GABA through action at a site proximal to the parallel fiber-basket cell synapse, at an as yet undetermined receptor. PMID- 10461858 TI - The effect of tryptophan plus methionine, 5-azacytidine, and methotrexate on adjuvant arthritis of rat. AB - Within the wider framework of our studies on the genesis of rheumatoid arthritis we have investigated the two signal processes in arthritis: adenoribosylation of proteins and DNA methylation. Arthritis can be induced when Freund's complete adjuvant is applied to rats. This form of arthritis can then be reduced or even totally suppressed through the application of several different substances. In the present article we have investigated if the effect of two of these substances, 5-azacytidine and methotrexate can be influenced by the application of tryptophan plus methionine. When applied singly, these latter two substances are known to reduce the formation of arthritis. This effect is intensified by a combination of tryptophan plus methionine. Application of tryptophan plus methionine without methotrexate or 5-azacytidine causes an enhanced development of an adjuvant induced arthritis. PMID- 10461859 TI - Nicotinamide and methionine reduce the liver toxic effect of methotrexate. AB - Methotrexate is widely used as a therapeutic agent in different diseases. This therapy is connected with various side effects, including liver toxicity. We have developed a mouse model to demonstrate the toxic effects of methotrexate: mice were given 50 mg/kg acetaminophen, which itself has no effect on the liver. If, additionally, methotrexate is applied, there is an increase in the death rate, as well as in glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) activities. If methotrexate is administered in conjunction with either nicotinamide or methionine, the rise in the death rate and in GOT and GPT activities associated with methotrexate application is markedly reduced. On the basis of these results, it can be concluded that methotrexate therapy should be combined with either nicotinamide or methionine, respectively. PMID- 10461860 TI - Antithrombotic effects of low-molecular-weight heparin calcium (LMWH-Ca) in experimental models. AB - The antithrombotic activity of low-molecular-weight heparin calcium (LMWH-Ca) was studied in venous and arterial thrombosis models, arterial thrombosis model in rats, arterio-venous shunt model and venous thrombosis model in rabbits. The data showed that LMWH-Ca reduced thrombus formation in a dose-dependent manner. It suggests that LMWH-Ca is a potent antithrombotic agent for venous thrombosis, and also may be a beneficial therapeutic agent in arterial thrombosis. PMID- 10461861 TI - Suppression of 125I-vasoactive intestinal polypeptide binding sites in arteries of the hamster seminal vesicle following castration. AB - The presence and distribution of 125I-vasoactive intestinal polypeptide (VIP) binding sites in blood vessels supplying the hamster seminal vesicle was studied using a receptor autoradiographic technique before and following castration. 125I VIP binding was studied in intact animals, in animals under a 15-day period of castration and in animals under the same period of castration but submitted to a further 15-day period of testosterone treatment. Our results show that, in the seminal vesicle, VIP-binding sites are localized in the gland smooth muscle coat and arterial smooth muscle. A 15-day castration period abolishes 125I-VIP binding to vascular smooth muscle but has no effect on 125I-VIP binding to the gland smooth muscle coat. Treatment with testosterone restores 125I-VIP binding to the vascular smooth muscle, completely reversing the effect of castration. Our results indicate that VIP-binding sites in the smooth muscle wall of arteries supplying the hamster seminal vesicle are under androgenic control and are more sensitive to androgen deprivation that VIP-binding sites associated to the gland smooth muscle coat. PMID- 10461862 TI - Myosin heavy chain profiles in regenerated fast and slow muscles innervated by the same motor nerve become nearly identical. AB - Plasticity of mature muscles exposed to different activation patterns is limited, probably due to restricted adaptive range of their muscle fibres. In this study, we tested whether satellite cells derived from slow muscles can give rise to a normal fast muscle, if transplanted to the fast muscle bed. Marcaine-treated rat soleus and extensor digitorum longus (EDL) muscles were transplanted to the EDL muscle bed and innervated by the 'EDL' nerve. Six months later expression of myosin heavy chain isoforms was analysed by areal densities of fibres, binding specific monoclonal antibodies, and by SDS gel electrophoresis. Both regenerated muscles closely resembled each other. Their myosin heavy chain profiles were similar to those in fast muscles although they were not identical to that in the control EDL muscle. Since not even regenerated EDL was able to reach the myosin heavy chain isoform profile of mature EDL muscle, our experimental model did not permit studying the adaptive capacity of satellite cells in different muscles in its whole extent. However, the results favour the multipotential myoblast stem cell population in rat muscles and underline the importance of the extrinsic regulation of muscle phenotype. PMID- 10461863 TI - Proteoglycan distribution pattern during aging in the nematode Caenorhabditis elegans: an ultrastructural histochemical study. AB - Glycosaminoglycans are important constituents of the extracellular matrix of vertebrates, where distinct changes in their distribution pattern occur during aging. However, little is known about their changes in the nematode Caenorhabditis elegans, which ages extremely rapidly compared to mammals. The presence of glycosaminoglycans was analysed in cross-sections of all organs of the nematode, in three different age groups (60, 144, 228 h), using the electron dense dye Cuprolinic Blue in conjunction with the critical electrolyte concentration method and specific glycosaminoglycan degrading enzymes. The nematodes (strain DH 26) were grown at 25.5 degrees C. The results indicate the presence of an organ-specific distribution pattern. Chondroitin-4-sulphate and/or chondroitin-6-sulphate are present in the epicuticula. Chondroitin-4-sulphate and/or chondroitin-6-sulphate and dermatan sulphate are detected in the mesocuticula. If stained by conventional methods the mesocuticula shows an empty fissure, which is filled by chondroitin sulphates and dermatan sulphate as shown by Cuprolinic Blue staining and enzymes. Heparan sulphate is found in the terminal web of intestinal cells while dermatan sulphate is revealed in the central cores of microvilli. An unknown polyanion staining at high electrolyte concentrations is observed in the gonads. Age-related changes do not impair the composition of the glycosaminoglycan fraction. In conclusion an unexpected highly differentiated pattern of glycosaminoglycans with high stability during aging exists. PMID- 10461864 TI - Histochemical and ultrastructural characterization of vacuoles and spherosomes as components of the lytic system in hyphae of the fungus Botrytis cinerea. AB - An integrated approach to acid phosphatase (EC 3.1.3.2) histochemistry by the azo dye and lead-capture ('Gomori') methods in phosphate-starved hyphae of the fungus Botrytis cinerea revealed strikingly different patterns of localization of activity staining. Reaction product formed with the azo-dye method was found in numerous small organelles (<0.5 microm diameter), which also accumulated the lipophilic dye Nile Red and mislocalized the formazan indicating mitochondrial succinate dehydrogenase activity. Such small organelles were stained only weakly and sporadically with the lead-capture method; instead, lead phosphate deposits were produced mainly in large vacuoles (up to 2.5 microm diam.), similar to those accumulating the vital dye Neutral Red. Additionally, acid phosphatase activity was detected in apical secretory vesicles with the lead-capture method but not with the azo-dye method. Ultrastructural studies by transmission electron microscopy confirmed the presence of large vacuoles which showed evidence of autophagic activity, and of small moderately osmiophilic organelles. The latter are considered to be spherosomes rather than lysosomes because of their weak reaction with the lead-capture method and their high lipid content. It is suggested that their apparently strong reaction with the azo-dye method is caused partly by false localization due to the lipophilic nature of the reaction product. PMID- 10461865 TI - An immunohistochemical study of macrophage influx and the co-localization of inducible nitric oxide synthase in the pancreas of non-obese diabetic (NOD) mice during disease acceleration with cyclophosphamide. AB - Cyclophosphamide has been used to accelerate and synchronize diabetes in non obese diabetic (NOD) mice. It was injected to 70-day-old female NOD mice and its effect on the progression of insulitis studied at days 0, 4, 7, 11 and at onset of diabetes. Pancreatic sections were also examined for the influx of CD4 and CD8 T cells and macrophages following immunofluorescence staining. The kinetics of macrophage immunoreactive cells in the exocrine and intra-islet areas were also investigated. Light and confocal microscopy were-employed to examine the expression and co-localization of inducible nitric oxide synthase following dual- and triple-label immunofluorescence histochemistry. After cyclophosphamide administration, the severity of insulitis remained similar from days 0 to 4 but began to rise at day 7 and markedly by day 11 and at onset of diabetes. At these two later stages, the insulitis scores were close to 100% while in age-matched control groups the insulitis scores were considerably lower. Immunohistochemical staining showed increasing numbers of CD4 and CD8 T cell subsets and macrophages within the islets and in exocrine, sinusoidal and peri-vascular regions. At onset of diabetes, several islets contained prominent clusters of macrophage immunoreactive cells. Macrophage influx into the islets increased sharply from day 7 (mean number per islet: 119 +/- 54 SEM), peaked at day 11 (mean number per islet: 228 +/- 42), and then declined at onset of diabetes (mean number per islet: 148 +/- 49). Several cells with immunolabelling for inducible nitric oxide synthase were detectable from day 7 onwards until the onset of diabetes. Dual- and triple-label immunohistochemistry showed that a significant proportion of macrophages and only a few beta cells contained the enzyme. Macrophages positive for the enzyme were located as clusters or occasionally contiguously, in the peri islet and intra-islet areas but rarely in the exocrine region. Islets with minimal distribution of macrophages in the peri-islet areas were not positive for inducible nitric oxide synthase. Beta cells positive for the enzyme were observed in islets with significant macrophage infiltration in locations close to macrophages. The present results show that cyclophosphamide administration to female NOD mice results in a rapid influx of CD4 and CD8 cells and macrophages. The marked up-regulation of inducible nitric oxide synthase in a selective proportion of macrophages, within the islets, immediately preceding and during the onset of diabetes suggests that nitric oxide released by islet macrophages may be an important molecular mediator of beta cell destruction in this accelerated model of insulin-dependent diabetes mellitus. PMID- 10461866 TI - Ultrastructural identification of caveolae and immunocytochemical as well as biochemical detection of caveolin in chondrocytes. AB - Using fluorescence immunocytochemistry, transmission electron microscopy and Western blotting, we have shown that caveolae and caveolin are abundant on chondrocytes of different cartilaginous structures of newborn and adult rat knee joints. Caveolin was detected in chondrocytes of the outer layer of articular cartilage, in the fibrocartilage of the menisci, and in fibrocartilage-like cells at tendon and ligament insertions. Electron microscopical studies revealed caveolae-like invaginations along the plasmalemmal membrane of articular chondrocytes and fibrocartilage cells. Immunoblot analysis demonstrated caveolin in detergent-insoluble and soluble complexes isolated from cultured rat chondrocytes. PMID- 10461867 TI - Persistent expression of developmental myosin heavy chain isoforms in the tapered ends of adult pigeon pectoralis muscle fibres. AB - We have shown previously that in addition to the adult myosin heavy chain (MyHC) isoform present throughout the length of each fast-twitch glycolytic muscle fibre within the pectoralis of the mature chicken, the neonatal isoform is retained in the tapered ends of these fibres. This work, however, has been the only published report of this phenomenon. Here, we tested the hypothesis that similar to the chicken, the ends of mature pigeon pectoralis muscle fibres contain developmental MyHC isoform(s). A histological stain was used to visualize endomysium to assist in the analysis of transverse sections of pectoralis muscle from four mature pigeons. Immunocytochemical techniques were used to localize MyHC isoform(s) characteristic of pigeon pectoralis development. We show that within mature pigeon pectoralis, the ends of both fast-twitch glycolytic and fast-twitch oxidative-glycolytic fibre types express MyHC isoform(s) characteristic of their earlier development. Thus, we extend our findings on chicken to another species and an additional muscle fibre type. Retention of developmental MyHC isoform(s) within the tapered ends of mature muscle fibres may be more widespread than is currently appreciated. PMID- 10461868 TI - Stage specific glycosylation pattern for lactoseries carbohydrates in the developing chick retina. AB - Based on the idea of differentiation-related changes in the glycosylation pattern of neurons, the expression of two cell surface oligosaccharide epitopes, N-acetyl lactosamine (NALA), and its sulpho-glucuronyl derivative (HNK-1), was studied, by immunohistochemistry and Western blot experiments, in the developing chick retina beginning on day 2 of incubation (E2) until day 18 post-hatching. NALA was detectable on neuroepithelial cells as soon as the primary optic vesicles formed, and this pattern continued until E3. During subsequent retinal development NALA expression became progressively restricted in concert with the appearance of postmitotic neurons as revealed by neurite outgrowth, and with the formation of synaptic contacts until it disappeared at the end of the incubation period. The pattern of NALA expression was the inverse of HNK-1 which was detected for the first time at E3 on postmitotic ganglion cells accumulating at the vitreal surface. The number of HNK-1+ cells steadily increased until around E10, when the entire neural epithelium was labelled. Synchronously to synaptogenesis, most neurons lost their HNK-1 immunoreactivity. At the time of hatching the adult-like pattern was found, characterised by subpopulations of labelled horizontal, bipolar, amacrine, and ganglion cells. Immunoblot experiments demonstrated transient NALA glycosylation of protein bands, partially identical in their apparent molecular weight to those proteins with HNK-1 glycosylation. The observed temporospatial changes in the glycosylation patterns of distinct proteins during retinal development suggest NALA as a suitable marker for neuronal proliferation, and HNK-1 for differentiation and establishment of final synaptic configuration. PMID- 10461869 TI - Chromosomal polymorphisms due to supernumerary chromosomes and pericentric inversions in the eyelidless microteiid lizard Nothobachia ablephara (Squamata, Gymnophthalmidae). AB - Cytogenetic studies were performed on eight specimens of the monotypic microteiid lizard Nothobachia ablephara, endemic of the sand dunes of the middle Sao Francisco river, in the semiarid caatinga, State of Bahia, Brazil. Chromosomes from fibroblast cultures were analysed after conventional, Ag-NOR staining, C-, and replication R- banding. A basic karyotype of 2n = 62, consisting mostly of subtelocentric and acrocentric chromosomes of decreasing size, was found in five specimens. Diploid number variation (2n = 63 and 2n = 64) occurred in two specimens due to the presence of one and two medium-sized subtelocentric supernumerary chromosomes (Bs). The Bs were not clearly distinguishable from the autosomes in Giemsa-stained metaphases and C-banding, but showed late replication after R-banding. Polymorphisms of pairs 1 and 5, observed in three different combinations, including acrocentrics, subtelocentrics, submetacentrics and metacentrics, were interpreted as the result of small pericentric inversions. Variation in the number of Ag-NORs was also reported. A chromosomal mechanism of sex determination of the XX:XY type is present in this species. Our data add more evidence to confirm the remarkable chromosomal variability that has been found in Gymnophthalmidae. PMID- 10461870 TI - Ribosomal DNA location in the scarab beetle Thorectes intermedius (Costa) (Coleoptera: Geotrupidae) using banding and fluorescent in-situ hybridization. AB - Mitotic metaphase chromosomes of the scarab beetle Thorectes intermedius (Costa) (Coleoptera Scarabaeoidea: Geotrupidae) were analyzed using various banding methods and fluorescent in-situ hybridization (FISH) with a ribosomal probe. The results obtained indicate that silver and CMA3 staining are unable to localize the chromosome sites of nucleolar organizer regions (NORs). Such an inadequacy is a consequence of the extensive silver and CMA3 stainability of both constitutive heterochromatin and heterochromatin associated to the NORs. PMID- 10461871 TI - Conservation of centromere protein in vertebrates. AB - The chicken genome comprises 78 chromosomes which include several macrochromosomes and many microchromosomes. Very little information is currently available concerning chicken centromere structure and function and it is unclear if the two types of chromosomes share a common centromere mechanism or whether this mechanism resembles those in other species. Immunofluorescence studies using antibodies to mammalian constitutive centromere proteins CENP-A, CENP-B, and CENP C and the passenger proteins CENP-E, and CENP-F revealed the presence of each of these proteins at the centromeres of both macro- and microchromsomes. CENP-A, CENP-B, and CENP-E levels showed variability between metaphase centromeres while CENP-C and CENP-F levels were relatively constant. These results suggest a common centromere mechanism for both types of chromosomes as well as indicating a high degree of conservation of individual proteins between widely divergent vertebrate classes and an overall conservation of centromere function throughout vertebrate evolution. PMID- 10461872 TI - Comparative genome analysis in American marsupials: chromosome banding and in situ hybridization. AB - We performed a comparative analysis of the G- and C-banded karyotypes of seven species of didelphid marsupials, representing the three diploid numbers (2n = 14, 18 and 22) known to occur in this family. In addition to a great similarity among karyotypes with the same diploid numbers, we also identified homeologies for all autosomal arms comprising the three karyotypes. Robertsonian rearrangements, pericentric inversions and heterochromatin variation account for the differences among the karyotypes. Interspecific variation in the size of the sex chromosomes is due to differences in heterochromatic content. In-situ hybridization with total genomic DNA revealed considerable conservation of the euchromatic portions of the three karyotypes and indicated divergence of repetitive DNA sequences in autosomal heterochromatin. PMID- 10461873 TI - X-irradiation of G1 CHO cells induces SCE which are both true and false in BrdU substituted cells but only false in biotin-dUTP-substituted cells. AB - The SCE-test is widely used in genetic toxicology and therefore knowledge of the contribution of BrdU to the formation of spontaneous and induced SCE is of great importance. The present study was undertaken to analyse the role of BrdU in X-ray induced SCE. If SCE resulted from inversions, rings and double minutes (RDM) would be the asymmetrical counterparts of SCE and should therefore have the same frequencies. Dose-effect relationships of SCE and RDM show that the frequencies of SCE are much higher than those of RDM. We conclude that only a few SCE may represent inversions. In a second set of experiments, endoreduplications were induced in cells irradiated either before or after labelling with BrdU. Analysis of SCE in endoreduplicated chromosomes allows the discrimination of the cell cycle in which they originated. The results show that SCE are only induced in the first cell cycle following irradiation of BrdU-substituted cells, indicating that labelling with BrdU is a necessary prerequisite for the formation of SCE. In order to test this directly, radiation-induced SCE frequencies were studied in cells prelabelled with BrdU or biotin-dUTP in a third set of experiments. The structure of biotin-dUTP suggests that, in contrast to BrdU, it does not give rise to radicals during irradiation. Significantly lower frequencies of SCE were observed in biotin-dUTP-substituted cells than in BrdU-labelled cells. Calculations show that nearly all SCE induced in biotin-dUTP-labelled chromosomes can be explained by chromosomal aberrations (false SCE). In contrast to this, most SCE induced by X-rays in BrdU-labelled cells are not due to chromosomal aberrations, but result from S-dependent lesions (true SCE). This clearly points towards radiation damage in BrdU-moieties as the source of DNA lesions leading to SCE. PMID- 10461874 TI - Comparative painting reveals strong chromosome homology over 80 million years of bird evolution. AB - Chickens and the great flightless emu belong to two distantly related orders of birds in the carinate and ratite subclasses that diverged at least 80 million years ago. In the first ZOO-FISH study between bird species, we hybridized single chromosome paints from the chicken (Gallus domesticus) onto the emu chromosomes. We found that the nine macrochromosomes show remarkable homology between the two species, indicating strong conservation of karyotype through evolution. One chicken macrochromosome (4) was represented by a macro- and a microchromosome in the emu, suggesting that microchromosomes and macrochromosomes are interconvertible. The chicken Z chromosome paint hybridized to the emu Z and most of the W, confirming that ratite sex chromosomes are largely homologous; the centromeric region of the W which hybridized weakly may represent the location of the sex determining gene(s). PMID- 10461875 TI - AFLPs represent highly repetitive sequences in Asparagus officinalis L. AB - The chromosomal and genomic organization of 5 cloned AFLP fragments in asparagus (Asparagus officinalis L.) were investigated. Two of the 5 AFLP loci were sex linked. The fragments, amplified with EcoRI/Mse I primers, ranged from 107 to 267 bp and were AT-rich. Southern hybridization gave interspersed, middle repetitive to high copy sequence signals. Fluorescence in-situ hybridization (FISH) exhibited hybridization signals on all chromosomes with dispersed distribution pattern and varying signal intensities. Repetitive signals in the form of clusters were observed on all chromosomes. In addition, the 5S rRNA gene was physically mapped on one pair of chromosomes and the 18S-5.8S-25S rRNA genes on three pairs. The results of the FISH and Southern analyses showed that the AFLP marker technology relies on repetitive sequences. Since repetitive DNA sequences represent a fraction of the plant genome undergoing rapid changes during the course of evolution, the question of whether such molecular markers originating from repetitive DNA sequences remain stable is discussed. PMID- 10461876 TI - Hypothesis: for the worst and for the best, L1Hs retrotransposons actively participate in the evolution of the human centromeric alphoid sequences. AB - A number of questions concerning the evolution and the function of the alpha satellite DNA sequences present at the centromere of all human chromosomes are still open. In this paper, we present data which could contribute to understanding these points. It is shown here that the alphoid sequences within which L1 elements are found are quite divergent from those of the homogeneous alphoid subsets present at each centromere where none has so far been detected. In addition, a number of L1s are detected close to the ends of the alpha satellite blocks. A fairly high proportion exhibit a polymorphism of presence/absence. Strikingly, several L1s localized at a distance from each other are always either present or absent simultaneously. This is interpreted as resulting from intrachromosomal recombination, through distant L1s, leading to deletion of several of them at once together with their surrounding alphoid sequences. The parameters determining which portion of the several megabases of alphoid sequences is actually involved in the centromeric function are not known. From the above data we suggest that the alpha satellite domain within which DNA sequences are recruited to form a centromere is both homogeneous in sequence and uninterrupted by L1s or any other retrotransposons. Conversely, non-centromere competent alphoid sequences would be both divergent and punctuated by scattered L1 elements, particularly at the borders of the alphoid blocks. On the grounds of these data and hypotheses, a model is presented in which it is postulated that accumulation of L1 insertions within a centromere competent alphoid domain is ruining this competence, the consequence being damage to or even loss of the centromere-forming capability of the chromosome. Restoration of fully centromere forming competence is supposed to occur by two alternative means, either de-novo amplification of a homogeneous and uninterrupted alphoid domain or by unequal crossing over with a homologue harbouring a large competent one. If L1 retrotransposons are acting detrimentally to centromere integrity (for the worst), one must also consider them as having positive consequences on chromosomes by preventing their centromeres from swelling indefinitely by the addition of alphoid sequences (for the best). The data and ideas presented here fit well with those already put forward by Csink and Henikoff (1998) using the example of Drosophila. PMID- 10461877 TI - Human HPA endoglycosidase heparanase. Map position 4q21.3. PMID- 10461878 TI - Glutathione and signal transduction in the mammalian CNS. AB - The tripeptide glutathione (GSH) has been thoroughly investigated in relation to its role as antioxidant and free radical scavenger. In recent years, novel actions of GSH in the nervous system have also been described, suggesting that GSH may serve additionally both as a neuromodulator and as a neurotransmitter. In the present article, we describe our studies to explore further a potential role of GSH as neuromodulator/neurotransmitter. These studies have used a combination of methods, including radioligand binding, synaptic release and uptake assays, and electrophysiological recording. We report here the characteristics of GSH binding sites, the interrelationship of GSH with the NMDA receptor, and the effects of GSH on neural activity. Our results demonstrate that GSH binds via its gamma-glutamyl moiety to ionotropic glutamate receptors. At micromolar concentrations GSH displaces excitatory agonists, acting to halt their physiological actions on target neurons. At millimolar concentrations, GSH, acting through its free cysteinyl thiol group, modulates the redox site of NMDA receptors. As such modulation has been shown to increase NMDA receptor channel currents, this action may play a significant role in normal and abnormal synaptic activity. In addition, GSH in the nanomolar to micromolar range binds to at least two populations of binding sites that appear to be distinct from all known excitatory amino acid receptor subtypes. GSH bound to these sites is not displaceable by glutamatergic agonists or antagonists. These binding sites, which we believe to be distinct receptor populations, appear to recognize the cysteinyl moiety of the GSH molecule. Like NMDA receptors, the GSH binding sites possess a coagonist site(s) for allosteric modulation. Furthermore, they appear to be linked to sodium ionophores, an interpretation supported by field potential recordings in rat cerebral cortex that reveal a dose-dependent depolarization to applied GSH that is blocked by the absence of sodium but not by lowering calcium or by NMDA or (S)-2-amino-3-hydroxy-5-methyl-4-isoxazolepropionate antagonists. The present data support a reevaluation of the role of GSH in the nervous system in which GSH may be involved both directly and indirectly in synaptic transmission. A full accounting of the actions of GSH may lead to more comprehensive understanding of synaptic function in normal and disease states. PMID- 10461879 TI - Characterization of the cDNA coding for rat brain cysteine sulfinate decarboxylase: brain and liver enzymes are identical proteins encoded by two distinct mRNAs. AB - Cysteine sulfinate decarboxylase (CSD) is considered as the rate-limiting enzyme in the biosynthesis of taurine, a possible osmoregulator in brain. Through cloning and sequencing of RT-PCR and RACE-PCR products of rat brain mRNAs, a 2,396-bp cDNA sequence was obtained encoding a protein of 493 amino acids (calculated molecular mass, 55.2 kDa). The corresponding fusion protein showed a substrate specificity similar to that of the endogenous enzyme. The sequence of the encoded protein is identical to that encoded by liver CSD cDNA. Among other characterized amino acid decarboxylases, CSD shows the highest homology (54%) with either isoform of glutamic acid decarboxylase (GAD65 and GAD67). A single mRNA band, approximately 2.5 kb, was detected by northern blot in RNA extracts of brain, liver, and kidney. However, brain and liver CSD cDNA sequences differed in the 5' untranslated region. This indicates two forms of CSD mRNA. Analysis of PCR amplified products of genomic DNA suggests that the brain form results from the use of a 3' alternative internal splicing site within an exon specifically found in liver CSD mRNA. Through selective RT-PCR the brain form was detected in brain only, whereas the liver form was found in liver and kidney. These results indicate a tissue-specific regulation of CSD genomic expression. PMID- 10461880 TI - Extensive alternative splicing in the 5'-untranslated region of the rat and human neuropeptide Y Y5 receptor genes regulates receptor expression. AB - Neuropeptide Y (NPY) elicits a plethora of physiological effects by interacting with several distinct G protein-coupled receptors. Activation of one of these receptors, the NPY Y5 receptor, is thought to result in increased food intake, anticonvulsant effects, attenuation of opiate withdrawal, inhibition of neuronal activity, and alteration of renal function. Several alternatively spliced human and rat NPY Y5 receptor cDNAs have been isolated that use different combinations of exons in the 5'-untranslated region. The various human NPY Y5 receptor cDNAs appear to be differentially expressed in different brain regions. The level of human NPY Y5 receptor expressed transiently in COS1 cells was significantly influenced by the sequence of the 5'-untranslated region. These results indicate that alternative splicing in the 5'-untranslated region of the human and rat NPY Y5 receptor genes occurs in a tissue-specific manner and is one mechanism by which cells control the level of NPY Y5 receptor expression. PMID- 10461881 TI - Regulation of synaptotagmin I phosphorylation by multiple protein kinases. AB - Synaptotagmin I has been suggested to function as a low-affinity calcium sensor for calcium-triggered exocytosis from neurons and neuroendocrine cells. We have studied the phosphorylation of synaptotagmin I by a variety of protein kinases in vitro and in intact preparations. SyntagI, the purified, recombinant, cytoplasmic domain of rat synaptotagmin I, was an effective substrate in vitro for Ca2+/calmodulin-dependent protein kinase II (CaMKII), protein kinase C (PKC), and casein kinase II (caskII). Sequencing of tryptic phosphopeptides from syntagI revealed that CaMKII and PKC phosphorylated the same residue, corresponding to Thr112, whereas caskII phosphorylated two residues, corresponding to Thr125 and Thr128. Endogenous synaptotagmin I was phosphorylated on purified synaptic vesicles by all three kinases. In contrast, no phosphorylation was observed on clathrin-coated vesicles, suggesting that phosphorylation of synaptotagmin I in vivo occurs only at specific stage(s) of the synaptic vesicle life cycle. In rat brain synaptosomes and PC12 cells, K+-evoked depolarization or treatment with phorbol ester caused an increase in the phosphorylation state of synaptotagmin I at Thr112. The results suggest the possibility that the phosphorylation of synaptotagmin I by CaMKII and PKC contributes to the mechanism(s) by which these two kinases regulate neurotransmitter release. PMID- 10461882 TI - Role of the outward delayed rectifier K+ current in ceramide-induced caspase activation and apoptosis in cultured cortical neurons. AB - We studied the novel hypothesis that an up-modulation of channels for outward delayed rectifier K+ current (I(K)) plays a key role in ceramide-induced neuronal apoptosis. Exposure for 6-10 h to the membrane-permeable C2-ceramide (25 microM) or to sphingomyelinase (0.2 unit/ml), but not to the inactive ceramide analogue C2-dihydroceramide (25 microM), enhanced the whole-cell I(K) current without affecting the transient A-type K+ current and increased caspase activity, followed by neuronal apoptosis 24 h after exposure onset. Tetraethylammonium (TEA) or 4-chloro-N,N-diethyl-N-heptylbenzenebutanaminium tosylate (clofilium), at concentrations inhibiting I(K), attenuated the C2-ceramide-induced caspase-3 like activation as well as neuronal apoptosis. Raising extracellular K+ to 25 mM similarly blocked the C2-ceramide-induced cell death; the neuroprotection by 25 mM K+ or TEA was not eliminated by blocking voltage-gated Ca2+ channels. An inhibitor of tyrosine kinases, herbimycin A (10 nM) or lavendustin A (0.1-1 microM), suppressed I(K) enhancement and/or apoptosis induced by C2-ceramide. It is suggested that ceramide-induced I(K) current enhancement is mediated by tyrosine phosphorylation and plays a critical role in neuronal apoptosis. PMID- 10461883 TI - Calnexin and the immunoglobulin binding protein (BiP) coimmunoprecipitate with AMPA receptors. AB - To identify proteins that interact with alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptors, we carried out coimmunoprecipitation analyses on detergent-solubilized rat forebrain membranes. Membranes were solubilized with Triton X-100, and immunoprecipitation was done using subunit specific antibodies to GluR1, GluR2/3, and GluR4 attached to protein Aagarose. Proteins bound to the antibodies were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by silver staining and western blotting. With solubilization in low ionic strength buffer, several coimmunoprecipitating proteins, with Mr = 17,000-100,000, were identified in silver-stained gels. Western blots were then probed with antibodies to a series of candidate proteins that were chosen based on the molecular masses of the copurifying proteins. Two of these were identified as the molecular chaperones calnexin (90 kDa) and the immunoglobulin binding protein (BiP; 78 kDa). Immunoprecipitation with antibodies to calnexin and BiP demonstrated that glycosylated AMPA receptor subunits were associated. The relationship between AMPA receptors and calnexin and BiP was further studied with immunocytochemistry of the hippocampus. Both calnexin and BiP labeling was present not only in the cell body but also in dendrites of hippocampal pyramidal neurons, where double label immunofluorescence also showed the presence of AMPA receptor subunits. PMID- 10461884 TI - Different roles for RhoA during neurite initiation, elongation, and regeneration in PC12 cells. AB - The goal of the present study was to characterize the effects of RhoA at different stages of nerve growth factor (NGF)-induced neuronal differentiation in the PC12 model. This comparative analysis was prompted by previous studies that reported apparently opposite effects for Rho in different models of neuronal differentiation and regeneration. PC12 cells were transfected with activated V14RhoA or dominant negative N19RhoA under the control of either a constitutive or a steroid-regulated promoter. Upon exposure to NGF, V14RhoA cells continued to proliferate and did not extend neurites; however, they remained responsive to NGF, as indicated by the activation of extracellular signal-regulated kinases. This inability to differentiate was reversed by C3 toxin and activation of cyclic AMP signaling, which inactivate RhoA. N19RhoA expression led to an increase in neurite initiation and branching. In contrast, when the RhoA mutants were expressed after NGF priming, only the rate of neurite extension was altered; V14RhoA clones had neurites approximately twice as long, whereas neurites of N19RhoA cells were approximately 50% shorter than those of appropriate controls. The effects of Rho in neurite regeneration mimicked those observed during the initial stages of morphogenesis; activation inhibited, whereas inactivation promoted, neurite outgrowth. Our results indicate that RhoA function changes at different stages of NGF-induced neuronal differentiation and neurite regeneration. PMID- 10461885 TI - Semaphorins as mediators of neuronal apoptosis. AB - Shrinkage and collapse of the neuritic network are often observed during the process of neuronal apoptosis. However, the molecular and biochemical basis for the axonal damage associated with neuronal cell death is still unclear. We present evidence for the involvement of axon guidance molecules with repulsive cues in neuronal cell death. Using the differential display approach, an up regulation of collapsin response mediator protein was detected in sympathetic neurons undergoing dopamine-induced apoptosis. A synchronized induction of mRNA of the secreted collapsin-1 and the intracellular collapsin response mediator protein that preceded commitment of neurons to apoptosis was detected. Antibodies directed against a conserved collapsin-derived peptide provided marked and prolonged protection of several neuronal cell types from dopamine-induced apoptosis. Moreover, neuronal apoptosis was inhibited by antibodies against neuropilin-1, a putative component of the semaphorin III/collapsin-1 receptor. Induction of neuronal apoptosis was also caused by exposure of neurons to semaphorin III-alkaline phosphatase secreted from 293EBNA cells. Anti-collapsin-1 antibodies were effective in blocking the semaphorin III-induced death process. We therefore suggest that, before their death, apoptosis-destined neurons may produce and secrete destructive axon guidance molecules that can affect their neighboring cells and thus transfer a "death signal" across specific and susceptible neuronal populations. PMID- 10461886 TI - Disruption of type IV intermediate filament network in mice lacking the neurofilament medium and heavy subunits. AB - To clarify the role of the neurofilament (NF) medium (NF-M) and heavy (NF-H) subunits, we generated mice with targeted disruption of both NF-M and NF-H genes. The absence of the NF-M subunit resulted in a two- to threefold reduction in the caliber of large myelinated axons, whereas the lack of NF-H subunits had little effect on the radial growth of motor axons. In NF-M-/- mice, the velocity of axonal transport of NF light (NF-L) and NF-H proteins was increased by about two fold, whereas the steady-state levels of assembled NF-L were reduced. Although the NF-M or NF-H subunits are each dispensable for the formation of intermediate filaments, the absence of both subunits in double NF-M; NF-H knockout mice led to a scarcity of intermediate filament structures in axons and to a marked approximately twofold increase in the number of microtubules. Protein analysis indicated that the levels of NF-L and alpha-internexin proteins were reduced dramatically throughout the nervous system. Immunohistochemistry of spinal cord from the NF-M-/-;NF-H-/- mice revealed enhanced NF-L staining in the perikaryon of motor neurons but a weak NF-L staining in axons. In addition, axonal transport studies carried out by the injection of [35S]methionine into spinal cord revealed after 30 days very low levels of newly synthesized NF-L proteins in the sciatic nerve of NF-M-/-;NF-H-/- mice. The combined results demonstrate a requirement of the high-molecular-weight subunits for the assembly of type IV intermediate filament proteins and for the efficient translocation of NF-L proteins into the axonal compartment. PMID- 10461887 TI - hFE65L influences amyloid precursor protein maturation and secretion. AB - The amyloid precursor protein (APP) is processed in the secretory and endocytic pathways, where both the neuroprotective alpha-secretase-derived secreted APP (APPs alpha) and the Alzheimer's disease-associated beta-amyloid peptide are generated. All three members of the FE65 protein family bind the cytoplasmic domain of APP, which contains two sorting signals, YTS and YENPTY. We show here that binding of APP to the C-terminal phosphotyrosine interaction domain of hFE65L requires an intact YENPTY clathrin-coated pit internalization sequence. To study the effects of the hFE65L/APP interaction on APP trafficking and processing, we performed pulse/chase experiments and examined APP maturation and secretion in an H4 neuroglioma cell line inducible for expression of the hFE65L protein. Pulse/chase analysis of endogenous APP in these cells showed that the ratio of mature to total cellular APP increased after the induction of hFE65L. We also observed a three-fold increase in the amount of APPs alpha recovered from conditioned media of cells overexpressing hFE65L compared with uninduced controls. The effect of hFE65L on the levels of APPs alpha secreted is due neither to a simple increase in the steady-state levels of APP nor to activation of the protein kinase C-regulated APP secretion pathway. We conclude that the effect of hFE65L on APP processing is due to altered trafficking of APP as it transits through the secretory pathway. PMID- 10461888 TI - The role of the 7B2 CT peptide in the inhibition of prohormone convertase 2 in endocrine cell lines. AB - Prohormone convertase (PC) 2 plays an important role in the processing of neuropeptide precursors via the regulated secretory pathway in neuronal and endocrine tissues. PC2 interacts with 7B2, a neuroendocrine protein that is cleaved to a 21-kDa domain involved in proPC2 maturation and a carboxyl-terminal peptide (CT peptide) that represents a potent inhibitor of PC2 in vitro. A role for the CT peptide as an inhibitor in vivo has not yet been established. To study the involvement of the CT peptide in PC2-mediated cleavages in neuroendocrine cells, we constructed a mutant proenkephalin (PE) expression vector containing PE with its carboxyl-terminal peptide (peptide B) replaced with the 7B2 inhibitory CT peptide. This PECT chimera was stably transfected into two PC2-expressing cell lines, AtT-20/PC2 and Rin cells. Although recombinant PECT proved to be a potent (nM) inhibitor of PC2 in vitro, cellular PC2-mediated cleavages of PE were not inhibited by the PECT chimera, nor was proopiomelanocortin cleavage (as assessed by adrenocorticotropin cleavage to alpha-melanocyte-stimulating hormone) inhibited further than in control cells expressing only the competitive substrate PE. Tests of stimulated secretion showed that both the CT peptide and the PE portion of the chimera were stored in regulated secretory granules of transfected clones. In both AtT-20/PC2 and Rin cells expressing the chimera, the CT peptide was substantially internally hydrolyzed, potentially accounting for the observed lack of inhibition. Taken together, our data suggest that overexpressed CT peptide derived from PECT is unable to inhibit PC2 in mature secretory granules, most likely due to its inactivation by PC2 or by other enzyme(s). PMID- 10461889 TI - Endothelins stimulate expression of cyclooxygenase 2 in rat cultured astrocytes. AB - Endothelin (ET) is one of the active endogenous substances regulating the functions of astrocytes. In the present study, we examined effects of ET on cyclooxygenase (COX) expression in cultured astrocytes. ET-3 (100 nM) caused transient increases in the expression of both COX2 mRNA and protein, but not those of COX1, in cultured astrocytes. ET-induced COX2 mRNA expression was suppressed by 5 microg/ml actinomycin D, 30 microM BAPTA/AM, inhibitors of protein kinase C (1-100 nM staurosporin and 100 microM H-7), 2 microM dexamethasone, and prolonged treatment with 100 nM phorbol 12-myristate 13 acetate. ET-3 stimulated production of prostaglandin (PG) E2 in cultured astrocytes. The effect of ET-3 on the PGE2 production was diminished by actinomycin D. Indomethacin and NS398, a selective COX2 inhibitor, comparably decreased both the basal and the ET-stimulated PGE2 production. Proliferation of cultured astrocytes was stimulated by 100 nM ET-3, and the increased proliferation was reduced by co-addition of 1 microM PGE2. Treatment with 1 microM PGE2 caused astrocytic morphological changes accompanied by disappearance of stress fibers, a prominent structure of organized cytoskeletal actin in cultured astrocytes. In the presence of 10 nM ET-3, PGE2 did not show an effect on astrocytic actin organization. The present study shows that ET is an inducer of astrocytic COX2 and suggests that ET-induced PGE2 production through COX2 may be involved in the regulation of astrocytic functions. PMID- 10461890 TI - Wnt signaling induces GLT-1 expression in rat C6 glioma cells. AB - The regulation of glial and neuronal Na+-dependent glutamate/aspartate transporters is of interest because abnormal glutamate transport may be responsible for certain neurological diseases. Because expression of the Wnt-1 protooncogene results in induction of the glial-type glutamate transporter GLAST in PC12 neuron-like cells, we have evaluated the effect of Wnt-1-induced signaling on glutamate transporter expression in rat C6 glioma cells. C6 cells are known normally to express EAAC1, a neuronal glutamate transporter, but not the GLAST or the GLT-1 glutamate transporter. C6 cells that ectopically expressed Wnt-1 contained a GLT-1 RNA species similar in size (>10 kb) to the GLT-1 transcript present in rat brain, and they also contained a previously unreported 3.3-kb GLT-1 RNA species. Both GLT-1 RNAs contain large parts of the coding region. However, the 3.3-kb GLT-1 species contains at least one small deletion within the coding region. The Wnt-1-expressing C6 cells contained little, if any, GLT-1 protein as determined by immunological techniques. We suggest that one or both of the GLT-1 RNA species induced by Wnt-1 either fail to be translated or yield abnormal translation products that are quickly degraded. Wnt-1-expressing C6 cells may thus represent a novel in vitro system for studying GLT-1 transporter expression at the transcriptional and/or posttranscriptional levels. PMID- 10461891 TI - Localization of the noradrenaline transporter in rat adrenal medulla and PC12 cells: evidence for its association with secretory granules in PC12 cells. AB - The noradrenaline transporter (NAT) is present in noradrenergic neurons and a few other specialized cells such as adrenal medullary chromaffin cells and the rat pheochromocytoma (PC12) cell line. We have raised antibodies to a 49-residue segment (NATM2) of the extracellular region (residues 184-232) of bovine NAT. Affinity-purified NATM2 antibodies specifically recognized an 80-kDa band in PC12 cell membranes by western blotting. Bands of a similar size were also detected in membranes from human neuroblastoma (SK-N-SH) cells expressing endogenous NAT and human embryonic kidney (HEK293) cells stably expressing bovine NAT. Immunocytochemistry of rat adrenal tissue showed that NAT staining was colocalized with tyrosine hydroxylase in medullary chromaffin cells. Most NAT immunoreactivity in rat adrenal chromaffin and PC12 cells was present in the cytoplasm and had a punctate appearance. Cell surface biotinylation experiments in PC12 cells confirmed that only a minor fraction of the NAT was present at the cell surface. Subcellular fractionation of PC12 cells showed that relatively little NAT colocalized with plasma membrane, synaptic-like microvesicles, recycling endosomes, or trans-Golgi vesicles. Most of the NAT was associated with [3H]noradrenaline-containing secretory granules. Following nerve growth factor treatment, NAT was localized to the growing tip of neurites. This distribution was similar to the secretory granule marker secretogranin I. We conclude that the majority of NAT is present intracellularly in secretory granules and suggest that NAT may undergo regulated trafficking in PC12 cells. PMID- 10461892 TI - Role of serotonin(2A) and serotonin(2B/2C) receptor subtypes in the control of accumbal and striatal dopamine release elicited in vivo by dorsal raphe nucleus electrical stimulation. AB - This study investigates, using in vivo microdialysis, the role of serotonin2A (5 HT2A) and 5-HT(2B/2C) receptors in the effect of dorsal raphe nucleus (DRN) electrical stimulation on dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), and 5-hydroxyindoleacetic acid (5-HIAA) extracellular levels monitored in the nucleus accumbens (NAC) and the striatum of halothane-anesthetized rats. Following DRN stimulation (300 microA, 1 ms, 20 Hz, 15 min) DA release was enhanced in the NAC and reduced in the striatum. The 5-HT2A antagonist SR 46349B (0.5 mg/kg) and the mixed 5-HT(2A/2B/2C) antagonist ritanserin (0.63 mg/kg) significantly reduced the effect of DRN stimulation on DA release in the NAC but not in the striatum. DA responses to DRN stimulation were not affected by the 5 HT(2B/2C) antagonist SB 206553 (5 mg/kg) in either region. None of these compounds was able to modify the enhancement of DOPAC and 5-HIAA outflow induced by DRN stimulation in either the NAC or the striatum. Finally, in both brain regions basal DA release was significantly increased only by SB 206553. These results indicate that 5-HT2A but not 5-HT(2B/2C) receptors participate in the facilitatory control exerted by endogenous 5-HT on accumbal DA release. Conversely, 5-HT(2B/2C) receptors tonically inhibit basal DA release in both brain regions. PMID- 10461893 TI - Nicotinic acetylcholine receptor antagonistic activity of monoamine uptake blockers in rat hippocampal slices. AB - The aim of our study was to investigate the effect of different monoamine uptake blockers on the nicotine-evoked release of [3H]noradrenaline ([3H]NA) from rat hippocampal slices. We found that desipramine (DMI), nisoxetine, cocaine, citalopram, and nomifensine inhibit the nicotine-evoked release of [3H]NA with an IC50 of 0.36, 0.59, 0.81, 0.93, and 1.84 microM, respectively. These IC50 values showed no correlation with the inhibitory effect (Ki) of monoamine uptake blockers on the neuronal NA transporter (r = 0.17, slope = 0.02), indicating that the NA uptake system is not involved in the process. In whole-cell patch clamp experiments neither drug blocked Na+ currents at 1 microM in sympathetic neurons from rat superior cervical ganglia, and only DMI produced a pronounced inhibition (52% decrease) at 10 microM. Comparison of the effect of DMI and tetrodotoxin (TTX) on the electrical stimulation- and nicotine-evoked release of [3H]NA showed that DMI, in contrast to TTX, inhibits only the nicotine-induced response, indicating that the target of DMI is not the Na+ channel. Our data suggest that monoamine uptake blockers with different chemical structure and selectivity are able to inhibit the nicotinic acetylcholine receptors in the CNS. Because these compounds are widely used in the therapy of depressed patients, our findings may have great importance in the evaluation of their clinical effects. PMID- 10461894 TI - Fluoxetine increases extracellular dopamine in the prefrontal cortex by a mechanism not dependent on serotonin: a comparison with citalopram. AB - Fluoxetine at 10 and 25 mg/kg increased (167 and 205%, respectively) the extracellular dopamine concentration in the prefrontal cortex, whereas 25 (but not 10) mg/kg citalopram raised (216%) dialysate dopamine. No compound modified dialysate dopamine in the nucleus accumbens. The effect of 25 mg/kg of both compounds on cortical extracellular dopamine was not significantly affected by 300 mg/kg p-chlorophenylalanine (PCPA) (fluoxetine, saline, 235%; PCPA, 230%; citalopram, saline, 179%; PCPA, 181%). PCPA depleted tissue and dialysate serotonin by approximately 90 and 50%, respectively, and prevented the effect of fluoxetine and citalopram on dialysate serotonin (fluoxetine, saline, 246%; PCPA, 110%; citalopram, saline, 155%; PCPA, 96%). Citalopram significantly raised extracellular serotonin from 0.1 to 100 microM (251-520%), whereas only 10 and 100 microM increased dialysate dopamine (143-231%). Fluoxetine similarly increased extracellular serotonin (98-336%) and dopamine (117-318%). PCPA significantly reduced basal serotonin and the effects of 100 microM fluoxetine (saline, 272%; PCPA, 203%) and citalopram (saline, 345%; PCPA, 258%) on dialysate serotonin but did not modify their effect on dopamine (fluoxetine, saline, 220%; PCPA, 202%; citalopram, saline, 191%; PCPA, 211%). The results clearly show that the effects of fluoxetine and of high concentrations of citalopram on extracellular dopamine do not depend on their effects on serotonin. PMID- 10461895 TI - Presynaptic kappa-opioid and muscarinic receptors inhibit the calcium-dependent component of evoked glutamate release from striatal synaptosomes. AB - In addition to cytosolic efflux, reversal of excitatory amino acid (EAA) transporters evokes glutamate exocytosis from the striatum in vivo. Both kappa opioid and muscarinic receptor agonists suppress this calcium-dependent response. These data led to the hypothesis that the calcium-independent efflux of striatal glutamate evoked by transporter reversal may activate a transsynaptic feedback loop that promotes glutamate exocytosis from thalamo- and/or corticostriatal terminals in vivo and that this activation is inhibited by presynaptic kappa and muscarinic receptors. Corollaries to this hypothesis are the predictions that agonists for these putative presynaptic receptors will selectively inhibit the calcium-dependent component of glutamate released from striatal synaptosomes, whereas the calcium-independent efflux evoked by an EAA transporter blocker, L trans-pyrrolidine-2,4-dicarboxylic acid (L-trans-PDC), will be insensitive to such receptor ligands. Here we report that a muscarinic agonist, oxotremorine (0.01-10 microM), and a kappa-opioid agonist, U-69593 (0.1-100 microM), suppressed the calcium-dependent release of glutamate that was evoked by exposing striatal synaptosomes to the potassium channel blocker 4-aminopyridine. The presynaptic inhibition produced by these ligands was concentration dependent, blocked by appropriate receptor antagonists, and not mimicked by the delta-opioid agonist [D-Pen2,5]-enkephalin. The finding that glutamate efflux evoked by L trans-PDC from isolated striatal nerve endings was entirely calcium independent supports the notion that intact basal ganglia circuitry mediates the calcium dependent effects of this agent on glutamate efflux in vivo. Furthermore, because muscarinic or kappa-opioid receptor activation inhibits calcium-dependent striatal glutamate release in vitro as it does in vivo, it is likely that both muscarinic and kappa receptors are inhibitory presynaptic heteroceptors expressed by striatal glutamatergic terminals. PMID- 10461896 TI - The kappa-opioid agonist, U-69593, decreases acute amphetamine-evoked behaviors and calcium-dependent dialysate levels of dopamine and glutamate in the ventral striatum. AB - The effects of a kappa-opioid receptor agonist on acute amphetamine-induced behavioral activation and dialysate levels of dopamine and glutamate in the ventral striatum were investigated. Amphetamine (2.5 mg/kg i.p.) evoked a substantial increase in rearing, sniffing, and hole-poking behavior as well as dopamine and glutamate levels in the ventral striatum of awake rats. U-69593 (0.32 mg/kg s.c.) significantly decreased the amphetamine-evoked increase in behavior and dopamine and glutamate levels in the ventral striatum. Reverse dialysis of the selective kappa-opioid receptor antagonist, nor-binaltorphimine, into the ventral striatum antagonized the effects of U-69593 on amphetamine induced behavior and dopamine and glutamate levels. Reverse dialysis of low calcium (0.1 mM) into the ventral striatum decreased basal dopamine, but not glutamate, dialysate levels by 91% 45 min after initiation of perfusion. Strikingly, 0.1 mM calcium perfusion significantly reduced the 2.5 mg/kg amphetamine-evoked increase in dopamine and glutamate levels in the ventral striatum, distinguishing a calcium-dependent and a calcium-independent component of release. U-69593 did not alter the calcium-independent component of amphetamine-evoked dopamine and glutamate levels. These data are consistent with the view that a transsynaptic mechanism augments the increase in dopamine and glutamate levels in the ventral striatum evoked by a moderately high dose of amphetamine and that stimulation of kappa-opioid receptors suppresses the calcium dependent component of amphetamine's effects. PMID- 10461897 TI - Activation of nuclear calcium dynamics by synaptic stimulation in cultured cortical neurons. AB - L-type voltage-sensitive Ca2+ channels (VSCCs) are enriched on the neuronal soma and trigger gene expression during synaptic activity. To understand better how these channels regulate somatic and nuclear Ca2+ dynamics, we have investigated Ca2+ influx through L-type VSCCs following synaptic stimulation, using the long wavelength Ca2+ indicator fluo-3 combined with laser scanning confocal microscopy. Single synaptic stimuli resulted in rapid Ca2+ transients in somatic cytoplasmic compartments (<5 ms rise time). Nuclear Ca2+ elevations lagged behind cytoplasmic levels by approximately 60 ms, consistent with a dependence on diffusion from a cytoplasmic source. Pharmacological experiments indicated that L type VSCCs mediated approximately 50% of the nuclear and somatic (cytoplasmic) Ca2+ elevation in response to strong synaptic stimulation. In contrast, relatively weak excitatory postsynaptic potentials (EPSPs; approximately 15 mV) or single action potentials were much less effective at activating L-type VSCCs. Antagonist experiments indicated that activation of the NMDA-type glutamate receptor leads to a long-lasting somatic depolarization necessary to activate L type VSCCs effectively during synaptic stimuli. Simulation of action potential and somatic EPSP depolarization using voltage-clamp pulses indicated that nuclear Ca2+ transients mediated by L-type VSCCs were produced by sustained depolarization positive to -25 mV. In the absence of synaptic stimulation, action potential stimulation alone led to elevations in nuclear Ca2+ mediated by predominantly non-L-type VSCCs. Our results suggest that action potentials, in combination with long-lived synaptic depolarizations, facilitate the activation of L-type VSCCs. This activity elevates somatic Ca2+ levels that spread to the nucleus. PMID- 10461898 TI - Effects of cannabinoids on dopamine release in the corpus striatum and the nucleus accumbens in vitro. AB - Cannabinoid receptors are widely distributed in the nuclei of the extrapyramidal motor and mesolimbic reward systems; their exact functions are, however, not known. The aim of the present study was to characterize the effects of cannabinoids on the electrically evoked release of endogenous dopamine in the corpus striatum and the nucleus accumbens. In rat brain slices dopamine release elicited by single electrical pulses was determined by fast cyclic voltammetry. Dopamine release was markedly inhibited by the OP2 opioid receptor agonist U 50488 and the D2/D3 dopamine receptor agonist quinpirole, indicating that our method is suitable for studying presynaptic modulation of dopamine release. In contrast, the CB1/CB2 cannabinoid receptor agonists WIN55212-2 (10(-6) M) and CP55940 (10(-6)-10(-5) M) and the CB1 cannabinoid receptor antagonist SR141716A (10(-6) M) had no effect on the electrically evoked dopamine release in the corpus striatum and the nucleus accumbens. The lack of a presynaptic effect on terminals of nigrostriatal and mesolimbic dopaminergic neurons is in accord with the anatomical distribution of cannabinoid receptors: The perikarya of these neurons in the substantia nigra and the ventral tegmental area do not synthesize mRNA, and hence protein, for CB1 and CB2 cannabinoid receptors. It is therefore unlikely that presynaptic modulation of dopamine release in the corpus striatum and the nucleus accumbens plays a role in the extrapyramidal motor and rewarding effects of cannabinoids. PMID- 10461899 TI - Regulation of myelin basic protein phosphorylation by mitogen-activated protein kinase during increased action potential firing in the hippocampus. AB - Myelin basic protein (MBP) phosphorylation is a complex regulatory process that modulates the contribution of MBP to the stability of the myelin sheath. Recent research has demonstrated the modulation of MBP phosphorylation by mitogen activated protein kinase (MAPK) during myelinogenesis and in the demyelinating disease multiple sclerosis. Here we investigated the physiological regulation of MBP phosphorylation by MAPK during neuronal activity in the alveus, the myelinated output fibers of the hippocampus. Using a phosphospecific antibody that recognizes the predominant MAPK phosphorylation site in MBP, Thr95, we found that MBP phosphorylation is regulated by high-frequency stimulation but not low frequency stimulation of the alveus. This change was blocked by application of tetrodotoxin, indicating that action potential propagation in axons is required. It is interesting that the change in MBP phosphorylation was attenuated by the reactive oxygen species scavengers superoxide dismutase and catalase and the nitric oxide synthase inhibitor N-nitro-L-arginine. Removal of extracellular calcium also blocked the changes in MBP phosphorylation. Thus, we propose that during periods of increased neuronal activity, calcium activates axonal nitric oxide synthase, which generates the intercellular messengers nitric oxide and superoxide and regulates the phosphorylation state of MBP by MAPK. PMID- 10461900 TI - Characterization of nucleotide transport into rat brain synaptic vesicles. AB - ATP transport to synaptic vesicles from rat brain has been studied using the fluorescent substrate analogue 1,N6-ethenoadenosine 5'-triphosphate (epsilon ATP). The increase in intravesicular concentration was time dependent for the first 30 min, epsilon-ATP being the most abundant nucleotide. The complexity of the saturation curve indicates the existence of kinetic and allosteric cooperativity in the nucleotide transport, which exhibits various affinity states with K0.5 values of 0.39 +/- 0.06 and 3.8 +/- 0.1 mM with epsilon-ATP as substrate. The Vmax values obtained were 13.5 +/- 1.4 pmol x min(-1) x mg of protein(-1) for the first curve and 28.3 +/- 1.6 pmol x min(-1) x mg of protein( 1) considering both components. This kinetic behavior can be explained on the basis of a mnemonic model. The nonhydrolyzable adenine nucleotide analogues adenosine 5'-O-3-(thiotriphosphate), adenosine 5'-O-2-(thiodiphosphate), and adenosine 5'-(beta,gamma-imino)triphosphate and the diadenosine polyphosphates P1,P3-di(adenosine)triphosphate, P1,P4-di(adenosine)tetraphosphate, and P1,P5 di(adenosine)pentaphosphate inhibited the nucleotide transport. The mitochondrial ATP/ADP exchange inhibitor atractyloside, N-ethylmaleimide, and polysulfonic aromatic compounds such as Evans blue and 4,4'-diisothiocyanostilbene-2,2' disulfonic acid also inhibit epsilon-ATP vesicular transport. PMID- 10461901 TI - Prion protein peptide neurotoxicity can be mediated by astrocytes. AB - A peptide based on amino acids 106-126 of the sequence of human prion protein (PrP106-126) is neurotoxic in culture. A role for astrocytes mediating PrP106-126 toxicity was investigated. The toxicity of PrP106-126 to cerebellar cell cultures was reduced by aminoadipate, a gliotoxin. Normally, PrP106-126 is not toxic to cultures containing neurones deficient in the cellular isoform of prion protein (PrPc). However, PrP106-126 was toxic to cerebellar cells derived from Prnp(0/0) mice (deficient in PrPc expression) when those cerebellar cells were cocultured with astrocytes. This toxicity was found to occur only in the presence of PrPc positive astrocytes and to be mediated by glutamate. Furthermore, PrPc-positive astrocytes were shown to protect Prnp(0/0) cerebellar cells from glutamate toxicity. This effect could be inhibited by PrP106-126. PrP106-126 did not enhance the toxicity of glutamate to neurones directly. When cerebellar cells were cocultured with astrocytes, the neurones became dependent on astrocytes for protection from glutamate toxicity and expressed an increased sensitivity to glutamate. In such a system, the protective effects of astrocytes against glutamate toxicity to neurones were inhibited by PrP106-126, resulting in a greater reduction in neuronal survival than would have been caused by PrP106-126 when astrocytes were not present. This new model provides a possible mechanism by which the gliosis in prion disease may accelerate the neurodegeneration seen in the later stages of the disease. PMID- 10461902 TI - Treatment of C6 glioma cells and rats with antidepressant drugs increases the detergent extraction of G(s alpha) from plasma membrane. AB - Results from previous studies suggested that chronic treatment of rats or C6 glioma cells with antidepressants augments the coupling between Gs and adenylyl cyclase. As these effects on C6 glioma cells are seen in the absence of presynaptic input, several antidepressant drugs may have a direct "postsynaptic" effect on their target cells. It was hypothesized that the target of antidepressant action was some membrane protein that may regulate coupling between G proteins and adenylyl cyclase. To test this, C6 glioma cells were treated with amitriptyline, desipramine, iprindole, or fluoxetine for 3 days. Chlorpromazine served as a control for these treatments. Membrane proteins were extracted sequentially with Triton X-100 and Triton X-114 from C6 glioma cells. Triton X-100 extracted more G(s alpha) in membranes prepared from antidepressant treated C6 glioma cells than from control groups. In addition, cell fractionation studies revealed that the amount of G(s alpha) in caveolin-enriched domains was reduced after antidepressant treatment and that adenylyl cyclase comigrated with G(s alpha) in the gradients. These data suggest that some postsynaptic component that increases availability of Gs to activate effector molecules, such as adenylyl cyclase, might be a target of antidepressant treatment. PMID- 10461903 TI - G Protein-coupled cyclic AMP signaling in postmortem brain of subjects with mood disorders: effects of diagnosis, suicide, and treatment at the time of death. AB - Components of cyclic AMP (cAMP) signaling were examined in postmortem cerebral cortex of a well characterized group of patients with mood disorders and nonpsychiatric control subjects. We measured G protein levels, adenylyl cyclase (AC) activity, and CREB levels in cerebral cortex of the subjects with respect to diagnosis, treatment, and suicide. There was no effect of diagnosis on any measure, except for a trend toward decreased stimulated AC activity in subjects with mood disorders relative to control subjects. We also detected a significant effect of suicide on temporal cortex CREB levels in subjects that died as a result of suicide relative to those that did not, which was more evident in patients with major depressive disorder. Bipolar disorder (BD) subjects treated with anticonvulsants at the time of death had decreased temporal cortex CREB levels relative to those not receiving anticonvulsants. Furthermore, we found a trend toward decreased occipital cortex G alpha(s) (short) levels in BD subjects treated with lithium. These results support the hypothesis of altered cAMP signaling in mood disorders and raise the possibility that factors other than diagnosis, such as treatment and suicide, may be relevant to cell-signaling abnormalities reported in the literature. PMID- 10461904 TI - Dopamine oxidation alters mitochondrial respiration and induces permeability transition in brain mitochondria: implications for Parkinson's disease. AB - Both reactive dopamine metabolites and mitochondrial dysfunction have been implicated in the neurodegeneration of Parkinson's disease. Dopamine metabolites, dopamine quinone and reactive oxygen species, can directly alter protein function by oxidative modifications, and several mitochondrial proteins may be targets of this oxidative damage. In this study, we examined, using isolated brain mitochondria, whether dopamine oxidation products alter mitochondrial function. We found that exposure to dopamine quinone caused a large increase in mitochondrial resting state 4 respiration. This effect was prevented by GSH but not superoxide dismutase and catalase. In contrast, exposure to dopamine and monoamine oxidase-generated hydrogen peroxide resulted in a decrease in active state 3 respiration. This inhibition was prevented by both pargyline and catalase. We also examined the effects of dopamine oxidation products on the opening of the mitochondrial permeability transition pore, which has been implicated in neuronal cell death. Dopamine oxidation to dopamine quinone caused a significant increase in swelling of brain and liver mitochondria. This was inhibited by both the pore inhibitor cyclosporin A and GSH, suggesting that swelling was due to pore opening and related to dopamine quinone formation. In contrast, dopamine and endogenous monoamine oxidase had no effect on mitochondrial swelling. These findings suggest that mitochondrial dysfunction induced by products of dopamine oxidation may be involved in neurodegenerative conditions such as Parkinson's disease and methamphetamine-induced neurotoxicity. PMID- 10461905 TI - Increased butyrylcholinesterase levels in microsomal membranes of dystrophic Lama2dy mouse muscle. AB - The proportions and the glycosylation of butyrylcholinesterase (BuChE) forms in vesicles rich in sarcoplasmic reticulum from normal (NMV) and dystrophic (DMV) muscle were analyzed, using merosin-deficient dystrophic mice. BuChE activity in DMV was two- to threefold that in NMV. Globular amphiphilic G1A, G2A, and G4A and hydrophilic G4H BuChE forms were identified in NMV and DMV. The amount of G2A forms increased sevenfold in DMV, and the other forms increased about twofold. The higher BuChE level in DMV might reflect a maturational defect, with dystrophy preventing the down-regulation of BuChE with muscle development. About half of G1A, G2A, and G4H BuChE forms in NMV or DMV bound to Lens culinaris agglutinin (LCA), a higher fraction to wheat germ agglutinin (WGA), and little to Ricinus communis agglutinin (RCA). Most of the G4A forms in NMV or DMV bound to LCA or WGA; those from NMV failed to bind to RCA, whereas most of the variants in DMV bound to it, suggesting that the excess of tetramers in DMV is mainly RCA reactive. The differential interaction of lectins with BuChE components from muscle microsomes, serum, and nerves confirmed that the microsomal BuChE was muscle-intrinsic. The results provide clues regarding the alterations that dystrophy produces in the biosynthesis of BuChE forms in muscle. PMID- 10461906 TI - Pharmacological characterization of morphine-induced in vivo release of cholecystokinin in rat dorsal horn: effects of ion channel blockers. AB - Previous studies indicate that an increased release of cholecystokinin (CCK) in response to morphine administration may counteract opioid-induced analgesia at the spinal level. In the present study we used in vivo microdialysis to demonstrate that systemic administration of antinociceptive doses of morphine (1 5 mg/kg, s.c.) induces a dose-dependent and naloxone-reversible release of CCK like immunoreactivity (CCK-LI) in the dorsal horn of the spinal cord. A similar response could also be observed following perfusion of the dialysis probe for 60 min with 100 microM but not with 1 microM morphine. The CCK-LI release induced by morphine (5 mg/kg, s.c.) was found to be calcium-dependent and tetrodotoxin sensitive (1 microM in the perfusion medium). Topical application of either the L type calcium channel blocker verapamil (50 microg) or the N-type calcium channel blocker omega-conotoxin GVIA (0.4 microg) onto the dorsal spinal cord completely prevented the CCK-LI release induced by morphine (5 mg/kg, s.c.). Our data indicate that activation of L- and N-type calcium channels is of importance for morphine-induced CCK release, even though the precise site of action of morphine in the dorsal horn remains unclear. The present findings also suggest a mechanism for the potentiation of opioid analgesia by L- and N-type calcium channel blocking agents. PMID- 10461907 TI - Enhancing effect of manganese on L-DOPA-induced apoptosis in PC12 cells: role of oxidative stress. AB - L-DOPA and manganese both induce oxidative stress-mediated apoptosis in catecholaminergic PC12 cells. In this study, exposure of PC12 cells to 0.2 mM MnCl2 or 10-20 microM L-DOPA neither affected cell viability, determined by the 3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, nor induced apoptosis, tested by flow cytometry, fluorescence microscopy, and the TUNEL technique. L-DOPA (50 microM) induced decreases in both cell viability and apoptosis. When 0.2 mM MnCl2 was associated with 10, 20, or 50 microM L-DOPA, a concentration-dependent decrease in cell viability was observed. Apoptotic cell death also occurred. In addition, manganese inhibited L-DOPA effects on dopamine (DA) metabolism (i.e., increases in DA and its acidic metabolite levels in both cell lysate and incubation medium). The antioxidant N-acetyl-L-cysteine significantly inhibited decreases in cell viability, apoptosis, and changes in DA metabolism induced by the manganese association with L-DOPA. An increase in autoxidation of L-DOPA and of newly formed DA is suggested as a mechanism of manganese action. These data show that agents that induce oxidative stress mediated apoptosis in catecholaminergic cells may act synergistically. PMID- 10461909 TI - Aberrant copper chemistry as a major mediator of oxidative stress in a human cellular model of amyotrophic lateral sclerosis. AB - We have investigated the response to oxidative stress in a model system obtained by stable transfection of the human neuroblastoma cell line SH-SY5Y with plasmids directing constitutive expression of either wild-type human Cu,Zn superoxide dismutase or a mutant of this enzyme (H46R) associated with familial amyotrophic lateral sclerosis. We report that expression of mutant H46R Cu,Zn superoxide dismutase induces a selective increase in paraquat sensitivity that is reverted by addition of D-penicillamine. Furthermore, expression of this mutant enzyme affects the activity of the endogenous wild-type enzyme both in basal conditions and in copper overloading experiments. Our data indicate that aberrant metal chemistry of this mutant enzyme is the actual mediator of oxidative stress and that concurrent impairment of the activity of wild-type endogenous enzyme compromises the cell's ability to respond to oxidative stress. PMID- 10461908 TI - Oxidative damage to the c-fos gene and reduction of its transcription after focal cerebral ischemia. AB - We investigated oxidative damage to the c-fos gene and to its transcription in the brain of Long-Evans rats using a transient focal cerebral ischemia and reperfusion (FCIR) model. We observed a significant (p < 0.001) increase in the immunoreactivity to 8-hydroxy-2'-guanine (oh8G) and its deoxy form (oh8dG) in the ischemic cortex at 0-30 min of reperfusion in all 27 animals treated with 15-90 min of ischemia. Treatment with a neuronal nitric oxide synthase (nNOS) inhibitor, 3-bromo-7-nitroindazole (60 mg/kg, i.p.), abolished the majority but not all of the oh8G/oh8dG immunoreactivity. Treatment with RNase A reduced the oh8G immunoreactivity, suggesting that RNA may be targeted. This observation was further supported by decreased levels of mRNA transcripts of the c-fos and actin genes in the ischemic core within 30 min of reperfusion using in situ hybridization. The reduction in mRNA transcription occurred at a time when nuclear gene damage, detected as sensitive sites to Escherichia coli Fpg protein in the transcribed strand of the c-fos gene, was increased 13-fold (p < 0.01). Our results suggest that inhibiting nNOS partially attenuates FCIR-induced oxidative damage and that nNOS or other mechanisms induce nuclear gene damage that interferes with gene transcription in the brain. PMID- 10461910 TI - Increase in external glutamate and NMDA receptor activation contribute to H2O2 induced neuronal apoptosis. AB - The present study aims to investigate the role of extracellular glutamate and NMDA receptor stimulation in the neuronal death induced by a transient exposure to H2O2 of cultured neurons originating from mouse cerebral cortex. Most of the neuronal loss following a transient exposure to H2O2 of cortical neurons results from an apoptotic process involving a secondary stimulation of NMDA receptors, which occurs after H2O2 washout. Indeed, (a) the neurotoxic effect of H2O2 was strongly reduced by antagonists of NMDA receptors, (b) the neurotoxic effect of H2O2 was enhanced in the absence of Mg2+, (c) the protective effect of MK-801 progressively decayed when it was applied with increasing delay time after H2O2 exposure, and (d), finally, the extracellular concentration of glutamate was increased after H2O2 exposure. The major part of H2O2-induced neurotoxicity is mediated by the formation of hydroxyl radicals, which might be involved in (a) the delayed accumulation of extracellular glutamate and NMDA receptor activation and (b) the poly(ADP-ribose) polymerase activation and the related NAD content decrease. The combination of these two mechanisms could lead to both an increase in ATP consumption and a decrease of ATP synthesis. The resulting large decrease in ATP content might be finally responsible for the neuronal death. PMID- 10461911 TI - Mitochondrial respiratory function and cell death in focal cerebral ischemia. AB - Pyruvate-supported oxygen uptake was determined as a measure of the functional capacity of mitochondria obtained from rat brain during unilateral middle cerebral artery occlusion and reperfusion. During ischemia, substantial reductions developed in both ADP-stimulated and uncoupled respiration in tissue from the focus of the affected area in the striatum and cortex. A similar pattern of change but with lesser reductions was seen in the adjacent perifocal tissue. Succinate-supported respiration was more affected than that with pyruvate in perifocal tissue at 2 h of ischemia, suggesting additional alterations to mitochondrial components in this tissue. Mitochondrial respiratory activity recovered fully in samples from the cortex, but not the striatum, within the first hour of reperfusion following 2 h of ischemia and remained similar to control values at 3 h of reperfusion. In contrast, impairment of the functional capacity of mitochondria from all three regions was seen in the first 3 h of reperfusion following 3 h of ischemia. Extensive infarction generally affecting the cortical focal tissue with more variable involvement of the perifocal tissue developed following 2 h of focal ischemia. Thus, mitochondrial impairment during the first 3 h of reperfusion was apparently not essential for tissue infarction to develop. Nonetheless, the observed mitochondrial changes could contribute to the damage produced by permanent focal ischemia as well as the larger infarcts produced when reperfusion was initiated following 3 h of ischemia. PMID- 10461912 TI - Calbindin D28K gene transfer via herpes simplex virus amplicon vector decreases hippocampal damage in vivo following neurotoxic insults. AB - Increases in cytoplasmic Ca2+ concentration ([Ca2+]i) can lead to neuron death. Preventing a rise in [Ca2+]i by removing Ca2+ from the extracellular space or by adding Ca2+ chelators to the cytosol of target cells ameliorates the neurotoxicity associated with [Ca2+]i increases. Another potential route of decreasing the neurotoxic impact of Ca2+ is to overexpress one of the large number of constitutive calcium-binding proteins. Previous studies in this laboratory demonstrated that overexpression of the gene for the calcium-binding protein calbindin D28K, via herpes simplex virus (HSV) amplicon vector, increases the survival of hippocampal neurons in vitro following energetic or excitotoxic insults but not following application of sodium cyanide. We now report that in vivo hippocampal infection with the calbindin D28K HSV vector increases neuronal survival in the dentate gyrus after application of the antimetabolite 3 acetylpyridine and increases transsynaptic neuronal survival in area CA3 following kainic acid neurotoxicity. The protective effects of infection with the calbindin D28K vector in an intact brain may prove to be beneficial during changes in Ca2+ homeostasis caused by neurological trauma associated with aging and certain neurological diseases. PMID- 10461913 TI - Blood-brain barrier produces significant efflux of L-aspartic acid but not D aspartic acid: in vivo evidence using the brain efflux index method. AB - The brain efflux index method has been used to clarify the mechanism of efflux transport of acidic amino acids such as L-aspartic acid (L-Asp), L-glutamic acid (L-Glu), and D-aspartic acid (D-Asp) across the blood-brain barrier (BBB). About 85% of L-[3H]Asp and 40% of L-[3H]Glu was eliminated from the ipsilateral cerebrum within, respectively, 10 and 20 min of microinjection into the brain. The efflux rate constant of L-[3H]Asp and L-[3H]Glu was 0.207 and 0.0346 min(-1), respectively. However, D-[3H]Asp was not eliminated from brain over a 20-min period. The efflux of L-[3H]Asp and L-[3H]Glu was inhibited in the presence of excess unlabeled L-Asp and L-Glu, whereas D-Asp did not inhibit either form of efflux transport. Aspartic acid efflux across the BBB appears to be stereospecific. Using a combination of TLC and the bioimaging analysis, attempts were made to detect the metabolites of L-[3H]Asp and L-[3H]Glu in the ipsilateral cerebrum and jugular vein plasma following a microinjection into parietal cortex, area 2. Significant amounts of intact L-[3H]Asp and L-[3H]Glu were found in all samples examined, including jugular vein plasma, providing direct evidence that at least a part of the L-Asp and L-Glu in the brain interstitial fluid is transported across the BBB in the intact form. To compare the transport of acidic amino acids using brain parenchymal cells, brain slice uptake studies were performed. Although the slice-to-medium ratio of D-[3H]Asp was the highest, followed by L-[3H]Glu and L-[3H]Asp, the initial uptake rate did not differ for both L-[3H]Asp and D-[3H]Asp, suggesting that the uptake of aspartic acid in brain parenchymal cells is not stereospecific. These results provide evidence that the BBB may act as an efflux pump for L-Asp and L-Glu to reduce the brain interstitial fluid concentration and act as a static wall for D-Asp. PMID- 10461914 TI - In vitro autoradiographic visualization of guanosine-5'-O-(3 [35S]thio)triphosphate binding stimulated by sphingosine 1-phosphate and lysophosphatidic acid. AB - Sphingosine 1-phosphate or lysophosphatidic acid activation of guanosine-5'-O-(3 [35S]thio)triphosphate ([35S]GTPgammaS) binding to G proteins was studied by in vitro autoradiography in rat and guinea pig brain. The highest stimulation of [35S]GTPgammaS binding by sphingosine 1-phosphate was observed in the molecular layer of the cerebellum. Marked stimulation was observed in most forebrain areas, including neocortex and striatum. With the exception of the substantia gelatinosa and nucleus of the solitary tract, sphingosine 1-phosphate-enhanced binding was weaker in the brainstem and spinal cord. Lysophosphatidic acid-enhanced labeling was only observed in white matter areas. The G protein inhibitor 5'-p fluorosulfonylbenzoyl guanosine completely inhibited lysophosphatidic acid enhanced [35S]GTPgammaS binding but only partially sphingosine 1-phosphate enhanced binding. N-Ethylmaleimide abolished binding stimulated by both agonists. Sphingosine 1-phosphate enhanced labeling by another GTP analogue (beta,gamma imido[8-3H]guanosine-5'-triphosphate) similarly to that of [35S]GTPgammaS. Lysophosphatidic acid stimulated [35S]GTPgammaS binding in the olfactory bulb, glia limitans, and cortical subventricular zone of 1-day-old rats, whereas enhanced labeling was not observed in the latter area of 5-day-old rats. Sphingosine 1-phosphate stimulated binding in the cortical and striatal subventricular zones and olfactory bulb in 1- and 5-day-old rats. In the absence of radioligand for sphingosine 1-phosphate and lysophosphatidic acid receptors, [35S]GTPgammaS autoradiography provides a unique opportunity to study the spatial distribution, ontogeny, and coupling properties of these receptors. PMID- 10461915 TI - The amino terminus with a conserved glutamic acid of G protein-coupled receptor kinases is indispensable for their ability to phosphorylate photoactivated rhodopsin. AB - To investigate functions of the consensus amino terminus of G protein-coupled receptor kinases (GRKs), two amino terminus-truncated mutants (delta30 or delta15) and two single-amino-acid mutants of conserved acidic residues (D2A or E7A) of human GRK1 were constructed and expressed in human embryonic kidney 293 cells. It was shown that truncated mutations and one single-point mutation (E7A) greatly decreased GRK1's activity to phosphorylate photoactivated rhodopsin (Rho*), whereas the abilities of these mutants to phosphorylate a synthetic peptide substrate and to translocate from cytosol to rod outer segments on light activation were unaffected. Further experiments demonstrated that the same truncated mutations (delta30 or delta15) of GRK2, representative of another GRK subfamily, also abolished the kinase's activity toward Rho*. The similar single point mutation (E5A) of GRK2 heavily impaired its phosphorylation of Rho* but did not alter its ability to phosphorylate the peptide, and the G329-rhodopsin augmented peptide phosphorylation by GRK2 (E5A) remained unchanged. Our data, taken together, suggest that the amino terminus as well as a conserved glutamic acid in the region of GRKs appears essential for their ability to functionally interact with G protein-coupled receptors. PMID- 10461916 TI - Light exposure activates retina ganglion cell lysophosphatidic acid acyl transferase and phosphatidic acid phosphatase by a c-fos-dependent mechanism. AB - We previously reported that the biosynthesis of phospholipids in the avian retina is altered by light stimulation, increasing significantly in ganglion cells in light and in photoreceptor cells in dark. In the present work, we have determined that light significantly increases the incorporation of [3H]glycerol into retina ganglion cell glycerophospholipids in vivo by a Fos-dependent mechanism because an oligonucleotide antisense to c-fos mRNA substantially blocked the light-dark differences. We also studied in vitro the enzyme activities of phosphatidate phosphohydrolase (PAPase), lysophosphatidate acyl transferase (AT II), and phosphatidylserine synthase from retinas of chickens exposed to light or dark. Higher PAPase I and AT II activities were found in incubations of retinal ganglion cells from animals exposed to light; no increase was observed in preparations obtained from light-exposed animals treated with the c-fos antisense oligonucleotide. No light-dark differences were found in phosphatidylserine synthase activity. These findings support the idea that a coordinated photic regulation of PAPase I and AT II is taking place in retina ganglion cells. This constitutes a reasonable mechanism to obtain an overall increased synthesis of glycerophospholipids in stimulated cells that is mediated by the expression of Fos-like proteins. PMID- 10461917 TI - Effect of G protein-coupled receptor kinase 2 on the sensitivity of M4 muscarinic acetylcholine receptors to agonist-induced internalization and desensitization in NG108-15 cells. AB - NG108-15 cells express predominantly the M4 subtype of the muscarinic receptor for acetylcholine. Stimulation of these receptors by the agonist carbachol causes an inhibition of cellular adenylyl cyclase and a consequent fall in the intracellular cyclic AMP concentration. Pretreatment of the cells with carbachol caused both internalization and desensitization of the M4 receptor. Overexpression of G protein-coupled receptor kinase (GRK) 2 caused an increase in the rate constant for receptor endocytosis (from 0.06 to 0.18 min(-1)) and a decrease in the EC50 for carbachol stimulation of internalization (from 15 to 3 microM). Overexpression of a dominant negative form of GRK2 had more modest effects, reducing the rate constant for endocytosis (from 0.11 to 0.07 min(-1)) and increasing the EC50 for carbachol stimulation of internalization (from 8 to 17 microM). Neither GRK2 nor dominant negative GRK2 overexpression had any effect on the rate constant for receptor recycling following agonist removal. The time course and extent of receptor desensitization in control cells were identical to the corresponding values for receptor internalization, and the rate and extent of desensitization were again increased by GRK2 overexpression. Exposure of the cells to hyperosmolar sucrose (0.6 M) almost completely blocked agonist-induced receptor internalization in both control and GRK2-overexpressing cells. Sucrose treatment also blocked agonist-induced desensitization. We conclude that the internalization and desensitization of the M4 muscarinic receptor in NG108-15 cells can be modulated in response to changes in GRK2 activity and also that internalization plays a key role in desensitization. PMID- 10461918 TI - The detoxification of cumene hydroperoxide by the glutathione system of cultured astroglial cells hinges on hexose availability for the regeneration of NADPH. AB - The ability of astroglia-rich primary cultures derived from the brains of newborn rats to detoxify exogenously applied cumene hydroperoxide (CHP) was analyzed as a model to study glutathione-mediated peroxide detoxification by astrocytes. Under the conditions used, 200 microM CHP disappeared from the incubation buffer with a half-time of approximately 10 min. The half-time of CHP in the incubation buffer was found strongly elevated (a) in cultures depleted of glutathione by a preincubation with buthionine sulfoximine, an inhibitor of glutathione synthesis, (b) in the presence of mercaptosuccinate, an inhibitor of glutathione peroxidase, and (c) in the absence of glucose, a precursor for the regeneration of NADPH. The involvement of glutathione peroxidase in the clearance of CHP was confirmed by the rapid increase in the level of GSSG after application of CHP. The restoration of the initial high ratio of GSH to GSSG depended on the presence of glucose during the incubation. The high capacity of astroglial cells to clear CHP and to restore the initial ratio of GSH to GSSG was fully maintained when glucose was replaced by mannose. In addition, fructose and galactose at least partially substituted for glucose, whereas exogenous isocitrate and malate were at best marginally able to replace glucose during peroxide detoxification and regeneration of GSH. These results demonstrate that CHP is detoxified rapidly by astroglial cells via the glutathione system. This metabolic process strongly depends on the availability of glucose or mannose as hydride donors for the regeneration of the NADPH that is required for the reduction of GSSG by glutathione reductase. PMID- 10461919 TI - A calcium/calmodulin-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate in the CNS of the cuttlefish Sepia officinalis: localization in specific neural pathways controlling the inking system. AB - Chemical, biochemical, and immunohistochemical evidence is reported demonstrating the presence in the brain of the cuttlefish Sepia officinalis of a Ca2+-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate, occurring in neurons and fibers functionally related to the inking system. Nitric oxide synthase activity was concentrated for the most part in the cytosolic fraction and was masked by other citrulline-forming enzyme(s). The labile nitric oxide synthase could be partially purified by ammonium sulfate precipitation of tissue extracts, followed by affinity chromatography on 2',5'-ADP-agarose and calmodulin agarose. The resulting activity, immunolabeled at 150 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis by antibodies to rat neuronal nitric oxide synthase, depended on NADPH and tetrahydro-L-biopterin, and was inhibited by N(G)-nitro-L-arginine. NMDAR2/3 subunit-immunoreactive proteins migrating at 170 kDa could also be detected in brain extracts, along with glutamate (whole brain: 0.32 +/- 0.03 micromol of glutamate/mg of protein; optic lobes: 0.22 +/- 0.04; vertical complex: 0.65 +/- 0.06; basal lobes: 0.58 +/- 0.04; brachial lobe: 0.77 +/- 0.06; pedal lobe: 1.04 +/- 0.08; palliovisceral lobe: 0.86 +/- 0.05). Incubation of intact brains with 1.5 mM glutamate or NMDA or the nitric oxide donor 2-(N,N-diethylamino)diazenolate-2-oxide caused a fivefold rise in the levels of cyclic GMP, indicating operation of the glutamate-nitric oxide-cyclic GMP signaling pathway. Immunohistochemical mapping of Sepia CNS showed specific localization of nitric oxide synthase-like and NMDAR2/3-like immunoreactivities in the lateroventral palliovisceral lobe, the visceral lobe, and the pallial and visceral nerves, as well as in the sphincters and wall of the ink sac. PMID- 10461920 TI - Synthesis and characterization of binding of 5-[76Br]bromo-3-[[2(S) azetidinyl]methoxy]pyridine, a novel nicotinic acetylcholine receptor ligand, in rat brain. AB - 5-[76Br]Bromo-3-[[2(S)-azetidinyl]methoxy]pyridine ([76Br]BAP), a novel nicotinic acetylcholine receptor ligand, was synthesized using [76Br]bromide in an oxidative bromodestannylation of the corresponding trimethylstannyl compound. The radiochemical yield was 25%, and the specific radioactivity was on the order of 1 Ci/micromol. The binding properties of [76Br]BAP were characterized in vitro and in vivo in rat brain, and positron emission tomography (PET) experiments were performed in two rhesus monkeys. In association experiments on membranes of the cortex and thalamus, >90% of maximal specific [76Br]BAP binding was obtained after 60 min. The dissociation half-life of [76Br]BAP was 51 +/- 6 min in cortical membranes and 56 +/- 3 min in thalamic membranes. Saturation experiments with [76Br]BAP revealed one population of binding sites with dissociation constant (K(D)) values of 36 +/- 9 and 30 +/- 9 pM in membranes of cortex and thalamus, respectively. The maximal binding site density (Bmax) values were 90 +/ 17 and 207 +/- 33 fmol/mg in membranes of cortex and thalamus, respectively. Scatchard plots were nonlinear, and the Hill coefficients were <1, suggesting the presence of a lower-affinity binding site. In vitro autoradiography studies showed that binding of [76Br]BAP was high in the thalamus and presubiculum, moderate in the cortex and striatum, and low in the cerebellum and hippocampus. A similar pattern of [76Br]BAP accumulation was observed by ex vivo autoradiography. In vivo, binding of [76Br]BAP in whole rat brain was blocked by preinjection of (S)(-)-nicotine (0.3 mg/kg) by 27, 52, 68, and 91% at survival times of 10, 25, 40, 120, and 300 min, respectively. In a preliminary PET study in rhesus monkeys, the highest [76Br]BAP uptake was found in the thalamus, and radioactivity was displaceable by approximately 60% with cytisine and by 50% with (S)(-)-nicotine. The data of this study indicate that [76Br]BAP is a promising radioligand for the characterization of nicotinic acetylcholine receptors in vivo. PMID- 10461921 TI - Processing of rat preprocortistatin in mouse AtT-20 cells. AB - Preprocortistatin (PPCST) has been recently identified as a novel somatostatin (SST)-related gene expressed only in brain. PPCST shares 11 of 14 residues with SST-14 at its C-terminal segment, where it features Lys-Lys and Lys-Arg basic sites for cleavage to putative cortistatin (CST)-14 and CST-29 peptides, respectively. Although synthetic replicates of the two putative CST peptides interact with SST receptors, they also display novel effects suggesting independent biological functions. Nothing is currently known about the naturally occurring mature cleavage products of PPCST posttranslational processing. Here we have cloned rat PPCST cDNA, stably expressed it in AtT-20 pituitary cells, and characterized the cellular and releasable products of PPCST processing by HPLC and radioimmunoassay using a SST-14 antibody that recognizes synthetic CST-14 and CST-29. Transfected cells released 120 +/- 21 pg of total CST-LI per plate basally, with an increase to 204 +/- 33 pg per plate with forskolin stimulation (p < 0.05). HPLC chromatograms of cell extracts revealed three peaks corresponding to CST-14, CST-29, and unprocessed PPCST (ratio, 41:55:4.5). CST was released preferentially as CST-14 (63-70%) compared with CST-29 (30-37%) under basal and forskolin-stimulated conditions. These studies demonstrate efficient processing of PPCST to both CST-14 and CST-29 through putative cleavage at both C-terminal dibasic sites of PPCST. Although the two peptides are synthesized approximately equally, CST-14 is released preferentially via the regulated secretory pathway. PMID- 10461922 TI - Regional distribution, ontogeny, purification, and characterization of the Ca2+ independent phospholipase A2 from rat brain. AB - We purified an 80-kDa Ca2+-independent phospholipase A2 (iPLA2) from rat brain using octyl-Sepharose, ATP-agarose, and calmodulin-agarose column chromatography steps. This procedure gave a 30,000-fold purification and yielded 4 microg of a near-homogeneous iPLA2 with a specific activity of 4.3 micromol/min/mg. Peptide sequences of the rat brain iPLA2 display considerable homology to sequences of the iPLA2 from P388D1 macrophages, Chinese hamster ovary cells, and human B lymphocytes. Under optimal conditions, the iPLA2 revealed the following substrate preference toward the fatty acid chain in the sn-2 position of phosphatidylcholine: linoleoyl > palmitoyl > oleoyl > arachidonoyl. The rat brain iPLA2 also showed a head group preference for choline > or = ethanolamine >> inositol. The iPLA2 is inactivated when exposed to pure phospholipid vesicles. The only exception is vesicles composed of phosphatidylcholine and phosphatidylinositol 4,5-bisphosphate. Studies on the regional distribution and ontogeny of various phospholipase A2 (PLA2) types in rat brain indicate that the iPLA2 is the dominant PLA2 activity in the cytosolic fraction, whereas the group IIA secreted PLA2 is the dominant activity in the particulate fraction. The activities of these two enzymes change during postnatal development. PMID- 10461923 TI - Cysteine 144 is a key residue in the copper reduction by the beta-amyloid precursor protein. AB - The beta-amyloid precursor protein (beta-APP) contains a copper-binding site localized between amino acids 135 and 156 (beta-APP(135-156)). We have employed synthetic beta-APP peptides to characterize their capacities to reduce Cu(II) to Cu(I). Analogues of the wild-type beta-APP(135-156) peptide, containing specific amino acid substitutions, were used to establish which residues are specifically involved in the reduction of copper by beta-APP(135-156). We report here that beta-APP's copper-binding domain reduced Cu(II) to Cu(I). The single-mutant beta APP(His147-->Ala) and the double-mutant beta-APP(His147-->Ala/His149-->Ala) showed a small decrease in copper reduction in relation to the wild-type peptide and the beta-APP(Cys144-->Ser) mutation abolished it, suggesting that Cys144 is the key amino acid in the oxidoreduction reaction. Our results confirm that soluble beta-APP is involved in the reduction of Cu(II) to Cu(I). PMID- 10461924 TI - Effect of social experience on dopamine-stimulated adenylyl cyclase activity and G protein composition in chick forebrain. AB - The stimulation of adenylyl cyclase (AC) by dopamine was investigated in membrane fractions of the forebrain areas mediorostral neostriatum/hyperstriatum ventrale (MNH) and lobus parolfactorius (LPO) of 8-day-old domestic chicks that had been raised under different social conditions: group A, socially isolated; group B, imprinted on an acoustic stimulus; group C, trained but nonimprinted; and group D, reared in small groups. Only in the brain of the socially experienced groups could cyclic AMP (cAMP) synthesis be stimulated by dopamine, but not in the socially isolated animals (group A). Ligand binding studies of dopamine D1- and D2-type receptors in membrane fractions did not reveal differences between socially experienced and isolated animals. Forskolin stimulation of total AC in MNH and LPO membrane fractions revealed a significantly enhanced AC stimulation in the socially reared but not in the imprinted group compared with isolated controls. Stimulation of AC by the G protein activator guanylylimidodiphosphate was significantly increased in the MNH and the LPO of socially reared chicks compared with isolated control animals. These results suggest that early postnatal social experience modulates the rate of cAMP synthesis and that these lasting changes are not due to changes of dopamine receptors but are related to increased AC activities and to increased sensitivity of Gs protein. PMID- 10461925 TI - Noninvasive measurements of [1-(13)C]glycogen concentrations and metabolism in rat brain in vivo. AB - Using a specific 13C NMR localization method, 13C label incorporation into the glycogen C1 resonance was measured while infusing [1-(13)C]glucose in intact rats. The maximal concentration of [1-(13)C]glycogen was 5.1 +/- 0.6 micromol g( 1) (mean +/- SE, n = 8). During the first 60 min of acute hyperglycemia, the rate of 13C label incorporation (synthase flux) was 2.3 +/- 0.7 micromol g(-1) h(-1) (mean +/- SE, n = 9 rats), which was higher (p < 0.01) than the rate of 0.49 +/- 0.14 micromol g(-1) h(-1) measured > or = 2 h later. To assess whether the incorporation of 13C label was due to turnover or net synthesis, the infusion was continued in seven rats with unlabeled glucose. The rate of 13C label decline (phosphorylase flux) was lower (0.33 +/- 0.10 micromol g(-1) h(-1)) than the initial rate of label incorporation (p < 0.01) and appeared to be independent of the duration of the preceding infusion of [1-(13)C]glucose (p > 0.05 for correlation). The results implied that net glycogen synthesis of approximately 3 micromol g(-1) had occurred, similar to previous reports. When infusing unlabeled glucose before [1-(13)C]glucose in three studies, the rate of glycogen C1 accumulation was 0.46 +/- 0.08 micromol g(-1) h(-1). The results suggest that steady-state glycogen turnover rates during hyperglycemia are approximately 1% of glucose consumption. PMID- 10461926 TI - Tyrosine hydroxylase is inactivated by catechol-quinones and converted to a redox cycling quinoprotein: possible relevance to Parkinson's disease. AB - Quinone derivatives of DOPA, dopamine, and N-acetyldopamine inactivate tyrosine hydroxylase, the initial and rate-limiting enzyme in the biosynthesis of the catecholamine neurotransmitters. The parent catechols are inert in this capacity. The effects of the catecholquinones on tyrosine hydroxylase are prevented by antioxidants and reducing reagents but not by scavengers of hydrogen peroxide, hydroxyl radicals, or superoxide radicals. Quinone modification of tyrosine hydroxylase modifies enzyme sulfhydryl groups and results in the formation of cysteinyl-catechols within the enzyme. Catecholquinones convert tyrosine hydroxylase to a redox-cycling quinoprotein. Quinotyrosine hydroxylase causes the reduction of the transition metals iron and copper and may therefore contribute to Fenton-like reactions and oxidative stress in neurons. The discovery that a phenotypic marker for catecholamine neurons can be converted into a redox-active species is highly relevant for neurodegenerative conditions such as Parkinson's disease. PMID- 10461927 TI - A novel effect of cyclic AMP on capacitative Ca2+ entry in cultured rat cerebellar astrocytes. AB - One of the most important intracellular Ca2+ regulatory mechanisms in nonexcitable cells, "capacitative Ca2+ entry" (CCE), has not been adequately studied in astrocytes. We therefore investigated whether CCE exists in cultured rat cerebellar astrocytes and studied the roles of cyclic AMP (cAMP) and protein kinase C (PKC) in CCE. We found that (1) at least two different intracellular Ca2+ stores, the endoplasmic reticulum and mitochondria, are present in cerebellar astrocytes; (2) CCE does exist in these cells and can be inhibited by Ni2+, miconazole, and SKF 96365; (3) CCE can be directly enhanced by an increase in intracellular cAMP, as 8-bromoadenosine 3',5'-cyclic monophosphate (8-brcAMP), forskolin, and isobutylmethylxanthine have stimulatory effects on CCE; and (4) neither of the two potent protein kinase A (PKA) inhibitors, H8 and H89, nor a specific PKA agonist, Sp-adenosine 3',5'-cyclic monophosphothioate, had a significant effect on cAMP-enhanced Ca2+ entry. The [Ca2+]i increase was not due to a release from calcium stores, hyperpolarization of the membrane potential, inhibition of calcium extrusion, or a change in pHi, suggesting that cAMP itself probably acts as a novel messenger to modulate CCE. We also conclude that activation of PKC results in an increase in CCE. cAMP and PKC seem to modulate CCE by different pathways. PMID- 10461928 TI - Malonate-induced generation of reactive oxygen species in rat striatum depends on dopamine release but not on NMDA receptor activation. AB - Intrastriatal injection of the reversible succinate dehydrogenase inhibitor malonate produces both energy depletion and striatal lesions similar to that seen in cerebral ischemia and Huntington's disease. The mechanisms of neuronal cell death involve secondary excitotoxicity and the generation of reactive oxygen species. Here, we investigated the effects of dopamine on malonate-induced generation of hydroxyl radicals and striatal lesion volumes. Using in vivo microdialysis, we found that malonate induced a 94-fold increase in extracellular striatal dopamine concentrations. This was paralleled by an increase in the generation of hydroxyl radicals. Prior unilateral lesioning of the nigrostriatal dopaminergic pathway by focal injection of 6-hydroxydopamine blocked the malonate induced increase in dopamine concentrations and the generation of hydroxyl radicals and attenuated the lesion volume. In contrast, the NMDA receptor antagonist MK-801 attenuated malonate-induced lesion volumes but did not block the generation of hydroxyl radicals. Thus, the dopaminergic and glutamatergic pathways are essential in the pathogenesis of malonate-induced striatal lesions. Our results suggest that the malonate-induced release of dopamine but not NMDA receptor activation mediates hydroxyl radical formation. PMID- 10461929 TI - Guidelines for the diagnosis and management of asthma: accepting the challenge. PMID- 10461930 TI - The impact of severe asthma on schoolchildren. AB - Episodic airway obstruction and hypoxia are potentially life-threatening to children with asthma and may account for neuropsychological impairment. Moreover, living with this chronic disease may severely disrupt children's emotional functioning. The general functioning of 25 children with severe asthma aged 10-13 years was tested by a comparison with 25 matched normal controls. Testing included variables with relevance to normal daily functioning: memory, concentration, school performance, physical condition, subjective symptoms after exercise, and negative emotions. The results showed that children with asthma did not significantly deviate from controls. They reported more dyspnea after physical exercise, which could not be attributed to lung function. Differences in school performance were not significant. It was concluded that children may generally adapt well to living with asthma. PMID- 10461931 TI - Relationship of flow and cross-sectional area to frictional stress in airway models of asthma. AB - Frictional stress from air flowing through narrowed airways may damage the airway mucosa and thereby increase airway inflammation and airway obstruction. To investigate the levels of frictional stress that might occur in the airway, we measured the frictional stress in three physical airway models (model 1: normal state; models 2 and 3: narrowed states with cross-sectional area half and one fourth of model 1, respectively) at tracheal expiratory flow rates of 1, 2, 3, 4, 5, 6, 7, and 8 L/sec. Frictional stress measured at stations in the trachea (A), two each in the left (B and C) and right (D and E) major bronchi, and one in the right secondary bronchus (F) indicated that at higher flow rates, high values of the frictional stress seen in model 3 (highest value being 139.2 N/m2 at 8 L/sec at station) could well damage the airway wall, especially during episodes of cough, and particularly when the mucosa is inflamed and friable as it is in asthmatic patients. Conversely, control of cough may have anti-inflammatory benefits in asthmatic patients. PMID- 10461932 TI - Physicians' approaches to providing asthma education to patients and the level of patient involvement in management decisions. AB - The objectives of this study were to describe physicians' self-reported approaches to providing disease-specific education to adults with asthma in an outpatient setting and their opinions about the level of patient involvement in management decisions. A mailed questionnaire was completed by 163 randomly selected physicians, representing an 80% response rate. The educational actions provided most frequently included information about prescribed medications (90% 100% of physicians), general asthma information (87%-98%), and inhaler demonstration (85%-95%). Educational activities provided least frequently were action plans (7%-74%) and referral to a nonprofit community asthma organization for further information (18%-36%). The reported provision of asthma education was related to patients' asthma severity (p < 0.0001) and physician specialty (p < 0.005). Physicians indicated that their patients were less involved in asthma management decisions than they would prefer (p < 0.001). The results suggest that physicians vary markedly in their approaches to providing asthma education to patients. Future descriptive and intervention studies are needed to identify the most effective models for providing education and patient involvement. PMID- 10461933 TI - Inhalation technique of 166 adult asthmatics prior to and following a self management program. AB - Self-management of asthma and self-treatment of exacerbations are considered important in the treatment of asthma. For successful self-treatment, medication has to be inhaled correctly, but the percentage of patients inhaling effectively varies widely. As part of a self-management program we checked and corrected inhalation technique. This paper addresses differences among inhalers in relation to patient characteristics and the effect of instruction, 1 year after enrollment. Maneuvers that are essential for adequate inhalation were identified. When errors in inhalation technique were observed, patients were instructed in the correct use of their devices. One year later, inhalation technique was checked again. Only patients who used the same inhaler throughout the entire study period were analyzed. Of the 245 adult asthmatic patients who were enrolled in the self-management program, 166 used the same inhaler throughout the study period. One hundred twenty patients (72%) performed all key items correctly at baseline and this increased to 80% after 1 year. At follow-up, older patients were less likely to demonstrate a perfect inhalation. Patients with a Diskhaler made fewest errors. Adjustment for differences in patient characteristics did not significantly change the results. Because many patients with asthma use their inhaler ineffectively, there is a need to know which inhaler leads to fewest errors. Diskhaler was nominated by this study. When patients are not able to demonstrate adequate inhalation technique in a "tranquil" setting, it is doubtful that they can do so when they experience an exacerbation. Therefore, inhalation instruction should be considered an essential ingredient, not only of self management programs, but also of asthma patient care in general. PMID- 10461934 TI - Requests for repeat medication prescriptions and frequency of acute episodes in asthma patients. AB - This study was conducted to determine if suboptimal use of inhaled steroid and over-reliance on bronchodilator medication to control asthma symptoms is associated with higher risk of acute asthma episodes. Details of repeat prescriptions for medication and use of health services over 12 months were collected for 754 adult outpatients with asthma; all were prescribed inhaled corticosteroid. Patients who requested less than five prescriptions per year were considered suboptimal users. Patients who requested seven or more bronchodilator prescriptions and less than five inhaled steroid prescriptions had significantly more family physician consultations for asthma episodes (p < 0.05), more hospital admissions (p < 0.05), and more disturbed nights in the week before hospital or family physician review (p < 0.05). Some patients with more severe asthma put themselves at risk by relying on bronchodilator medication rather than regular inhaled steroid for asthma control. Among patients who were low bronchodilator users, those who requested few inhaled steroid prescriptions were younger and more anxious but did not have an increased risk of acute asthma episodes. PMID- 10461935 TI - Item responsiveness of a rhinitis and asthma symptom score during a pollen season. AB - Twenty-one asthma patients with allergic rhinitis completed a series of self administered questionnaires (21-item symptom score for rhinosinusitis and asthma, bother scale, McMaster Asthma Quality of Life Questionnaire [MAQOL] and Euroqol) at 2-week intervals from August to November 1994. Relative responsiveness of the instruments was assessed in reference to the maximum and minimum average scores for MAQOL, with area under the curve (AUC) and correlation coefficients between the different instruments. Symptom score, MAQOL, and bother scale provided similar results for both extreme values and AUC, whereas Euroqol utilities were less responsive. These results suggest that the symptom scores and bother scales are responsive and valid, and might prove valuable in everyday practice, clinical trials, and quality assurance programs. PMID- 10461936 TI - Circadian variation in exhaled nitric oxide in nocturnal asthma. AB - Asthma is characterized by airway inflammation and shows a circadian variation with nocturnal exacerbations. Because exhaled nitric oxide (ENO) measurement appears to be a noninvasive marker of airway inflammation, we examined the hypothesis that ENO would increase at night. In five nocturnal and five non nocturnal asthmatics, ENO was measured at 4 P.M., 10 P.M., and 4 A.M. before and after bronchodilator. Both pre- and post-bronchodilator ENO (mean pre- and post bronchodilator +/- SEM, ppb) unexpectedly fell significantly in nocturnal asthma from 4 P.M. (77.2 +/- 8.2) compared to 10 P.M. (68.4 +/- 8.7, p < 0.003) and 4 A.M. (66.0 +/- 8.5, p < 0.001) with no significant difference between 10 P.M. and 4 A.M.. In contrast, there were no significant differences in mean ENO at 4 P.M., 10 P.M., and 4 A.M. in non-nocturnal asthma. (51.3 +/- 10.8, 57.7 +/- 13.4, 53.8 +/- 12.5 ppb, respectively). Following bronchodilator, ENO rose significantly by 10.5 +/- 1.8 ppb in the nocturnal asthma group alone. The circadian rhythm of ENO differed greatly between nocturnal and non-nocturnal asthma. The significant decrease in ENO in nocturnal asthma may reflect an important chronobiological defect in the endogenous production and/or increased disposition of nitric oxide, which in view of its bronchodilator action, could play a role in nocturnal exacerbations of asthma. PMID- 10461938 TI - Arginine kinase and phosphoarginine, a functional phosphagen, in the rhabditoid nematode steinernema carpocapsae. AB - Moderate activity of arginine kinase was found in Steinernema carpocapsae, an entomopathogenic nematode. In the forward reaction, 4.60 and 3.12 micromol ATP/min/mg protein was produced in infectious third-stage juveniles (J3s) and adult nematodes, respectively. For the reverse reaction, 3.20 and 2.27 micromol phosphoarginine/min/mg protein was produced by J3s and adults, respectively. The K(m)s for phosphoarginine and ADP were 0.73 and 0.42 mM, respectively, in the forward reaction, whereas in the reverse reaction, the K(m)s were 0.37 and 2.35 mM for arginine and ATP, respectively, for the enzyme from J3s. The pH optimum for the forward reaction was 7.2 and 7.3 in J3s and adults, respectively. The pH optimum was elevated for the reverse reaction, 7.8 and 7.9-8.5 in J3s and adults, respectively. In the J3s, the in vitro optima for arginine kinase activity was correlated with the in vivo tissue pH in hypoxic (6.9) and aerobic (7.5) J3s estimated by in vivo flow 31P-NMR. PMID- 10461937 TI - Citric-acid cycle key enzyme activities during in vitro growth and metacyclogenesis of Leishmania infantum promastigotes. AB - The activities of 5 key regulatory enzymes in most energetic systems, namely citrate synthase (EC 4.1.3.7, CS), NADP-specific isocitrate dehydrogenase (EC 1.1.1.42, ICDH), succinate dehydrogenase (EC 1.3.99.1, SDH), L-malate dehydrogenase (EC 1.1.1.37, MDH), and decarboxylating malic enzyme (EC 1.1.1.40, ME), were measured during the growth and metacyclogenesis of a cutaneous (CL) and a visceral (VL) strain of Leishmania infantum. As occurs with other Leishmania species, infective promastigotes were present along all phases of growth, but their percentages were higher at the early stationary phase for VL and the end of the same phase for CL. High CS and SDH activities were detected in both strains, as compared with other trypanosomatids, bringing more evidence for an actively functional citric-acid cycle in L. infantum. Both strains showed higher levels of CS, ICDH, and MDH and lower SDH and ME activities when more metacyclic promastigotes were present, but in VL these changes paralleled an increase in glucose consumption, whereas in CL these changes coincided with an NH3 hyperproduction. This suggests that the energy metabolism during L. infantum growth and metacyclogenesis is affected by regulated enzymes that probably respond to changes in the culture medium in the levels of glucose and amino acids. PMID- 10461939 TI - The ultrastructure of gametogenesis of cryptosporidium baileyi (eimeriorina; cryptosporidiidae) in the respiratory tract of broiler chickens (Gallus domesticus). AB - The ultrastructural features of sexual development of Cryptosporidium baileyi in the respiratory tract of experimentally infected broiler chickens were studied using transmission electron microscopy. Sexual stages of C. baileyi were seen attached to the tracheal epithelium and free in the tracheal lumen. These stages included intracellular type III merozoite-like stages, microgamonts, microgametes, macrogamonts, thin-walled oocysts, and thick-walled oocysts. These stages were developmentally similar to those observed for other Cryptosporidium species. All of the above stages were observed during each study day. Thin-walled oocysts, microgamonts, and microgametes were seen less frequently than other sexual stages. Microgamonts, macrogamonts, and oocysts attached to the epithelium were all contained in a host cell membrane or within a parasitophorous vacuole. Thin-walled oocysts of C. baileyi were observed for the first time on an ultrastructural level in the respiratory tract of chickens. PMID- 10461940 TI - The morphogenesis of Ascaris suum to the infective third-stage larvae within the egg. AB - Studies of the morphology of Ascaris suum larvae developing in the egg during embryonation in vitro at room temperature showed that 2 molts take place within the egg. The first larval stage (L1) appeared in the egg after 17-22 days of cultivation, the first molt to the second larval stage (L2) took place from day 22 to day 27, and the second molt to the third larval stage (L3) started on day 27 and continued during the 60-day observation period. Infectivity of the eggs was studied by oral egg inoculation in mice and showed that the L3 are the infective stage for mice. Molting to the L3 stage occurs gradually over a period of 2-6 wk, and it is recommended to have an additional maturation period so the infectivity of an egg batch may reach maximum level. PMID- 10461941 TI - Molecular genetic key for the identification of 17 Ixodes species of the United States (Acari:Ixodidae): a methods model. AB - A taxonomic key, based on restriction enzyme analysis of the second internal transcribed spacer (ITS-2) in the nuclear ribosomal DNA gene, was developed for identification of 17 Ixodes tick species in the United States. This key includes: Ixodes affinis Neumann, Ixodes angustus Neumann, Ixodes baergi Cooley and Kohls, Ixodes brunneus Koch, Ixodes cookei Packard, Ixodes dentatus Marx, Ixodes jellisoni Cooley and Kohls, Ixodes kingi Bishopp, Ixodes minor Neumann, Ixodes muris Bishopp and Smith, Ixodes pacificus Cooley and Kohls, Ixodes scapularis Say, Ixodes sculpularis Neumann, I. spinipalpis Hadwen and Nuttall, Ixodes texanus Banks, Ixodes uriae White, and Ixodes woodi Bishopp. A 900-bp fragment of the ITS-2 was amplified using the polymerase chain reaction. This fragment was then digested with the restriction enzymes MspI and CfoI, and the digested fragments were size fractionated on a 2.5% high-resolution agarose gel. A dichotomous key was developed based on digested fragment sizes relative to a standard set of size markers. Little intraspecific variation in restriction fragment banding patterns was detected. PMID- 10461942 TI - Ultrastructure of shizonts and merozoites of Sarcocystis falcatula in the lungs of budgerigars (Melopsittacus undulatus). AB - Transmission electron microscopy was used to study the ultrastructure of schizogony of Sarcocystis falcatula in the lungs of budgerigars (Melopsittacus undulatus). Schizogony occurred exclusively by endopolygeny within endothelial cells of pulmonary capillaries, venules, and small veins. Early schizonts were elongate with a large nucleus and nucleolus, surrounded by a pellicle consisting of a plasmalemma and an inner single membrane, and contained most of the organelles and inclusion bodies found in merozoites of Sarcocystis species. As development proceeded, schizonts increased in size and conformed to the shapes of the pulmonary blood vessels. As micronemes, dense granules, the conoid, and subpellicular microtubules disappeared, there was an increase in the size and number of mitochondria, Golgi complexes, and Golgi adjuncts (apicoplasts). As the nucleus elongated, there was a progressive increase in the number of spindles located at various intervals along the nuclear envelope. Eventually, 2 merozoites formed internally immediately above each spindle. During endopolygeny, a portion of the nucleus was incorporated into each merozoite bud along with 1 or 2 Golgi adjuncts, a Golgi complex, mitochondria, endoplasmic reticulum, and ribosomes. During merozoite formation, micronemes appeared in close association with the Golgi complex and gradually increased in number. The pellicle invaginated around the merozoites so they budded at the schizont surface leaving behind a small, central residual body. Dense granules appeared after merozoites were completely formed. Schizonts were 24 x 6.8 microm and contained 24-96 merozoites. Merozoites were 5.1 x 1.8 microm and were found free in the pulmonary air passages and pulmonary capillaries and within nearly all cells of the lung except red blood cells. PMID- 10461943 TI - Evolutionary relationships among the protostrongylidae (Nematoda: Metastrongyloidea) as inferred from morphological characters, with consideration of parasite-host coevolution. AB - The phylogeny of nematodes in the family Protostrongylidae (Nematoda: Metastrongyloidea) was reconstructed by cladistic analysis of 28 binary and multistate characters derived from comparative morphology. Analyses were hierarchical, and examined (1) relationships among genera, including 13 ingroup taxa and Metastrongylidae as an outgroup (single tree, 78 steps, consistency index [CI] = 0.705); and (2) relationships among genera and species groups, including 21 ingroup taxa and Metastrongylus apri as an outgroup (single tree, 76 steps, CI = 0.582). In the species-level tree, Protostrongylidae was divided into 2 major clades, 1 containing the subfamilies Muelleriinae (including the recently described Umingmtakstrangylus pallikuukensis), Elaphostrongylinae, and the Varestrongylinae (excluding Pneumocaulus kadenazii). Varestrongylus was paraphyletic as it included Pneumostrongylus calcaratus. The second major clade consisted of a paraphyletic group containing Protostrongylus spp. and Spiculocaulus leuckarti and, basal to this subclade, several other individual protostrongylid lineages. The various subclades generally correspond to the subfamilial divisions of the Protostrongylidae. The Neostrongylinae, however, is not supported as Neostrongylus and Orthostrongylus are not sister groups. Based on a large number of hypothesized synapomorphies, the elaphostrongylines appear to be a highly derived group of protostrongylids, a feature potentially correlated with their habitat localization in muscular and nervous tissues. The generic-level tree retained most of the primary structure revealed among the species but excluded the varestrongylines from the Muelleriinae + Elaphostrongylinae subclade. Artiodactyles of the family Cervidae are considered basal hosts for protostrongylids; secondary colonization in Caprini, Rupicaprini, and among lagomorphs is postulated. PMID- 10461944 TI - Nucleotide sequence variability in the nontranscribed spacer of the rRNA locus in the oyster parasite Perkinsus marinus. AB - We examined the sequence variability of the nontranscribed spacer (NTS) and internal-transcribed spacer (ITS1 and ITS2) domains of the rRNA locus of Perkinsus marinus from Maryland, Florida, and Louisiana. The sequence of P. marinus DNA including the 5S rRNA, NTS, small subunit (SSU) rRNA, ITSI, and ITS2 regions confirmed their contiguity in the rRNA locus and revealed differences at 28 positions with the SSU rRNA sequences published earlier. The 307-bp polymerase chain reaction (PCR)-amplified fragments from the NTS domain of the various P. marinus isolates revealed the presence of 2 distinct sequences, designated as types I and II, that differed at 6 defined nucleotide positions. Based on these differences, nested PCR and restriction enzyme digests were used to distinguish between the 2 types. Sequences of the ITS1 and ITS2 domains of samples from either NTS type I (n = 3) or type II (n = 3) showed no variation and were identical to published sequences. Frequencies of the P. marinus NTS sequence types I and II in infected oysters varied with the geographic origin of the samples. All Maryland samples examined (n = 19) corresponded to the NTS type I sequence, the type II was the most frequent in the Florida samples (n = 17), and both types were about equally represented in the Louisiana samples (n = 19), with both sequence types found in individual oyster specimens. Although it has been suggested that P. marinus is diploid, it remains to be determined if both NTS sequence types can be present in a single P. marinus trophozoite. PMID- 10461945 TI - Infection and immunity with the RH strain of Toxoplasma gondii in rats and mice. AB - Infection and immunity to toxoplasmosis induced by the RH strain of Toxoplasma gondii was compared in Sprague-Dawley (SD) and Wistar rats and in outbred Swiss Webster mice. All rats injected with up to 1,000,000 RH-strain tachyzoites remained clinically normal, whereas mice injected with only 1 live tachyzoite died of acute toxoplasmosis. Rats could be infected with 1 tachyzoite of the RH strain as shown by antibody development and by bioassay in mice. However, after 8 days, RH-strain organisms were recovered only inconsistently from SD and Wistar rat brains. Contrary to a report of sterile immunity to T. gondii infection in rats after immunization with live RH tachyzoites, we found infection immunity after challenge with the VEG strain. Toxoplasma gondii tissue cysts of the VEG strain could be recovered from most SD and Wistar rats, first injected with live RH-strain tachyzoites and then challenged with oocysts of the VEG strain. Our RH strain, and probably many others, passed for 50+ yr as tachyzoites has lost not only the capacity to form oocysts, but also shows a marked reduction or absence of tissue cyst (bradyzoites) formation. PMID- 10461946 TI - Immunological characterization of antigens released by Trypanosoma cruzi-infected cells. AB - Chagas' disease, caused by Trypanosorna cruzi, is characterized by the appearance of pathological lesions in the heart and other tissues during the chronic phase. The mechanisms responsible for such damage are still unclear. In the vertebrate host, T. cruzi replicates intracellularly before transforming from amastigotes into trypomastigotes. The infected host cell then lyses, releasing the cytoplasmic contents and the parasites that shed membrane glycoproteins soon after release. The sum of all these components we have termed released antigen (Rag). We characterized antigens, released in vitro by fibroblasts infected with T. cruzi, obtained by concentrating conditioned serum-free culture media. The results demonstrate that Rag contains a complex protein mixture including stage specific T. cruzi antigens (Ssp-1, -2, -4), glucose-regulated protein (Grp) 78h, and peptides recognized by the monoclonal antibody 2B10. These peptides exhibit neuraminidase activity and are expressed by intracellular and 10-20% of released trypomastigotes. Additionally, Rag is recognized by sera from T. cruzi-infected mice and human chagasic patients. Rag also stimulates in vitro production of interferon-gamma by splenocytes from resistant C57B1/6 and susceptible BALB/c infected mice and interleukin-4 by splenocytes from BALB/c infected mice. Altogether these results indicate that Rag is immunologically relevant and could contribute to pathogenesis of T. cruzi infection. PMID- 10461947 TI - Adaptation of the AMRU-1 strain of Plasmodium vivax to Aotus and Saimiri monkeys and to four species of anopheline mosquitoes. AB - A chloroquine-resistant strain of Plasmodium vivax (AMRU-1) from Papua New Guinea has been adapted to grow in 4 species of Aotus monkeys (Aotus lemurinus griseimembra, Aotus vaciferans, Aotus nancymai, and Aotus azarae boliviensis), hybrid Aotus monkeys, and Saimiri boliviensis monkeys. Whereas it was possible to infect Saimiri monkeys with this parasite by inoculation of parasitized erythrocytes, only 42% of Saimiri monkeys became infected, compared to 92% of Aotus monkeys attempted. Comparative mosquito feedings showed that only A. vociferans, A. l. griseimembra, and Saimiri boliviensis monkeys produced infections in mosquitoes. Oocysts were observed on the guts of the 4 species of mosquitoes used (Anopheles gambiae, Anopheles stephensi, Anopheles freeborni, and Anopheles dirus), but sporozoite transmission was effected only with the intravenous inoculation of sporozoites from An. dirus into an A. l. griseimembra monkey. PMID- 10461948 TI - The fate of Cryptosporidium parvum oocysts ingested by dung beetles and their possible role in the dissemination of cryptosporidiosis. AB - The fate of oocysts of Cryptosporidium parvum ingested by dung beetles and the possible role these beetles serve in the dissemination of cryptosporidiosis were tested on the following species: Anoplotrupes stercorosus, Aphodius rufus, and Onthophagus fracticornis. Ten specimens of each species were offered cattle dung supplemented with 5.9 x 10(6) oocysts of C. purvum. After 24 hr of feeding, the beetles were examined for the presence of oocysts on their external surfaces, in their gastrointestinal tracts, and in feces passed during the experiment. Results indicate that although many oocysts pass safely through the mouthparts and gastrointestinal tracts of the beetles, the majority of them are destroyed. Coprophagous insects can, therefore, be considered an important aspect in the ecology of gastrointestinal diseases of man and livestock, as both agents of control and dissemination. PMID- 10461949 TI - Possible role of the 34-kilodalton hyaluronic acid-binding protein in visceral Leishmaniasis. AB - Leishmania donovani, the causative organism of human visceral leishmaniasis, invades host macrophages through its interaction with the cell surface molecules of target cells. The presence of a cell surface protein (Mr 34 kDa) having specific affinity toward hyaluronan (HA), a major extracellular matrix component, has been previously reported in macrophage cell lines. In order to identify the possible role of this HA-binding protein (HABP) in leishmaniasis, initially we demonstrated its overexpression in spleen, liver, macrophages, and serum of hamsters infected with L. donovani. We further observed higher levels of HABP in the macrophage cell line J774.G8 upon infection with L. donovani. Finally, we observed a significant increase in the level of HABP in the serum of patients with kala-azar. In order to understand its functional role in leishmaniasis, we report here a significant inhibition of cellular phosphorylation of HABP in hamster macrophages infected with L. donovani. Interestingly, the 34-kDa HABP was shown to bind with 2 proteins of promastigotes as well as amastigotes of L. donovani (with molecular masses of 55 kDa and 30 kDa respectively), suggesting a possible role for HABP in adhesion during the interaction of promastigotes and macrophages. PMID- 10461950 TI - Serological differences in Neospora caninum-associated epidemic and endemic abortions. AB - A sandwich enzyme-linked immunosorbent assay (ELISA) for the sensitive and specific detection of bovine antibodies to Neospora caninum was developed and evaluated using sera from cattle experimentally infected with N. caninum, Toxoplasma gondii, Sarcocystis cruzi, Sarcocystis hominis, Sarcocystis hirsuta, Eimeria bovis, Cryptosporidium parvum, Babesia divergens, and field sera from naturally exposed animals. Field sera were classified using a gold standard that included the results from an indirect fluorescent antibody test (IFAT) and an immunoblot (IB). Based on these gold standard results, i.e., IFAT-IB results, an equal relative sensitivity and specificity of 94.2%(theta0) was reached when a cutoff of 0.034 (d0) was employed. The analysis of IFAT-IB-positive field sera showed that within groups of aborting and nonaborting dams, the animals from herds with endemic N. caninum-associated abortions had significantly higher ELISA indices than animals from herds with N. caninum-associated epidemic abortions. By contrast, IFAT-IB-positive aborting dams from herds with endemic N. caninum associated abortions had significantly lower IFAT titers than IFAT-IB-positive aborting dams from herds with epidemic N. caninum-associated abortions. This is the first time that statistically significant serological differences between herds exhibiting epidemic and endemic N. caninum-associated abortions are described. PMID- 10461951 TI - A cladistic approach to relationships in pentastomida. AB - The positioning of the Pentastomida among the Metazoa has provoked many debates up to the present. On the other hand, the internal relationships among the pentastomid subgroups have received much less attention in the past. We provide the first phylogenetic analyses under Hennigian principles. Thirty-two morphological characters were selected from the primary literature, analyzed manually, and then with the program Hennig86. Four most parsimonious trees were obtained; these were analyzed by successive weighting and reduced to 1 consensus cladogram 380 steps long, with a consistency index of 0.98 and a retention index of 0.99. Characters were also analyzed as unordered, producing results that were congruent with the previous analyses. The internal groups were ordered according to the following system: (Heymnonsicambria + Haffnericambria + Backlericambria (Cephalobaenida (Railietiellida nov. (Reighardiida nov. (Porocephalida (Linguatuloidea (Linguatulidae + Subtriquetridae) + Porocephaloidea (Sebekidae + Porocephalidae))))))). This phylogenetic system is largely congruent with the first modern taxonomic arrangement proposed for the Pentastomida. PMID- 10461952 TI - New species of Spinuris (monogenea: monocotyladae) from Zapteryx exasperata (elasmobranchii: rhinobatidae) from Baja California Sur, Mexico. AB - Monocotylid monogeneans were collected from the gills of 2 Rhinobatos productus and 5 Zapteryx exasperata in Bahia Almeja, Baja California Sur, Mexico. All are parasitized by monocotylids of the genus Spinuris, 9 specimens of Spinuris lophosoma Doran on Rhinobatos productus (new record) and 27 specimens of Spinuris zapterygis n. sp. on Z. exasperata. This new species differs from the other species in the genus Spinuris by the number of dorsal haptoral sclerites, morphology of hooks, and male copulatory apparatus. The generic diagnosis of the genus Spinuris is revised. PMID- 10461954 TI - Reintroduction of Profilicollis Meyer, 1931, as a genus in acenthocephala: significance of the intermediate host. AB - Polymorphid acanthocephalans with long necks, spheroid proboscides, and eggs without polar swellings of the fertilization membrane constitute a morphologically distinct group of species. In the past, this taxon was considered a separate genus, Profilicollis. More recently, workers have been unwilling to accept these features as generic-level characters, and presently the group is considered a subgenus of Polymorphus. An analysis prompted by our studies of various polymorphid cystacanths in crabs along the coasts of Scotland and the U.S.A. reveals that all records of Polymorphus in decapods refer to species with 6 cement glands frequently assigned to Hexaglandula, to incidental occurrences of species purported to be Polymorphus minutus, or to species of the subgenus Profilicollis. Occurrence in decapod crustaceans implies substantial life history differences from the other species of the genus occurring in amphipod crustaceans. We conclude this, together with morphological distinctiveness, justifies return of Profilicollis to full generic status within Polymorphidae. PMID- 10461953 TI - Molecular differentiation and phylogeny of entomopathogenic nematodes (rhabditida: heterorhabditidae) based on ND4 gene sequences of Mitochondrial DNA. AB - We determined partial ND4 gene sequences of mitochondrial DNA from 15 heterorhabditid nematode isolates, representing 5 species collected from different regions of the world, by using polymerase chain reaction (PCR) and direct-sequencing of PCR products. Aligned nucleotide as well as amino acid sequences were used to differentiate nematode species by comparing sequence divergence and to infer phylogeny of the nematodes by using maximum parsimony and likelihood methods. Robustness of our phylogenetic trees was checked by bootstrap tests. The 15 nematode isolates can be divided into 7 haplotypes based on DNA sequences. On a larger scale, the sequence divergence revealed 4 distinct groups corresponding to 4 described species. No sequence divergence was detected from 5 isolates of Heterorhabditis bacteriophora or between Heterorhabditis marelatus to Heterorhabditis hepialius. Our sequence data yielded phylogenetic trees with identical topologies when different tree-building methods were used. Most relationships were also confirmed by using amino acid sequences in maximum parsimony analysis. Our molecular phylogeny of Heterorhabditis species support an existing taxonomy that is based largely on morphology and the sequence divergence of the ND4 gene permits species identification. PMID- 10461955 TI - Two new species of coccidia (apicomplexa: Eimeriidae) from the bearded false chameleon Chamaeleolis barbatus (Sauria: polychridae) from cinco pesos, Pinar Del Rio, Cuba. AB - Parasitological examination of bearded false chameleons Chamaeleolis barbatus freshly imported from Cuba revealed the presence of 2 species of coccidia that are described as new. Oocysts of Isospora chamaeleolidis n. sp. are spherical to slightly subspherical, 16.1 (13-21) x 15.6 (13-19) microm, with a brownish and bilayered wall approximately 1.0-1.5 microm thick; outer layer markedly pitted. 0.75-1.0 microm thick. One, rarely 2, globular polar granules, 1.5 in diameter are present in the sporulated oocysts. Sporocysts are ellipsoidal, 10.8 (10-13) x 7.8 (7-9) microm, with a smooth, colorless, and unilayered sporocyst wall. Stieda body and substieda bodies are present. A sporocyst residuum is present, consisting of small granules of irregular size scattered among the sporozoites. Oocysts of Eimeria chamaeleolidisbarbati n. sp. are broadly oval, 19.0 (17-21) x 15.7 (15-17) microm, with a bilayered, colorless oocyst wall approximately 0.75 thick; outer layer of oocyst wall is smooth, 0.5 microm thick. One or 2, rarely 4, globular, irregular polar granules, approximately 1.5 microm in diameter, are present in sporulated oocysts. Sporocysts are broadly oval, 7.4 (7-8.5) x 6.1 (5.5-7) microm, with a smooth, colorless, and unilayered sporocyst wall, composed of 2 valves joined by suture; Stieda body and substieda bodies are absent. PMID- 10461956 TI - Activity of atovaquone against Babesia microti in the Mongolian gerbil, Meriones unguiculatus. AB - The hydroxynaphthoquinone, atovaquone (Wellvone, Glaxo-Wellcome Ltd.) was found to have significant activity against Babesia microti, the main cause of human babesiosis in the U.S.A. This activity compares well with that of the most effective babesicide currently available for use in animals, imidocarb dipropionate, that unlike atovaquone is not licensed for use in humans. Treatment with well tolerated doses of atovaquone results in a rapid reduction in parasitemias and an early disappearance of parasites from blood smears. However, in common with all the other babesicides tested, atovaquone did not sterilize gerbils of infection, even at very high daily doses administered for up to 10 days. A combination of atovaquone and clindamycin was more effective than atovaquone alone in the treatment of both acute and chronic infections but failed to eliminate parasites completely. PMID- 10461957 TI - Biological effects of lithium chloride on Herpetomonas muscarum muscarum and Blastocrithidia culicis (kinetoplastida: trypanosomatidae). AB - Lithium is widely used in medicine as an antidepressive drug and for myelosuppression attenuation during chemotherapy. In spite of abundant literature, questions on the biological action of lithium ions are far from being answered. We have here examined the effects of lithium (10-200 mM) on culture forms of the trypanosomatid protozoa Herpetornonas muscarum muscarum and Blastocrithidia culicis. Incubation of these parasites with LiCl inhibited cell growth in a concentration-dependent manner, but growth could be restored when the drug was removed from the medium. Furthermore, Li+ induced cell differentiation in H. m. muscarum. Light microscopy examination of cell viability, using erythrosin B staining, showed that all treated parasites remained viable with all drug concentrations used. Ultrastructural analysis by transmission electron microscopy showed that the cells presented no signs of degeneration. However, in H. m. muscarum the nuclei lost their peripheral heterochromatin and appeared filled with a homogeneous matrix, whereas in B. culicis an increased amount of lipid droplets was present in the cytoplasm. Our data show that LiCl treatment arrested the cell division process, stimulated cell differentiation, and affected the metabolism of these parasites. PMID- 10461958 TI - Schistosoma mansoni: IL-4 is necessary for concomitant immunity in mice. AB - To ask whether type-2 immune responses serve an essential role in concomitant immunity, that is the prevention of superinfection with Schistosoma mansoni, we compared resistance to a challenge infection in infected wild-type (WT) mice and in infected IL-4-/- mice, which are unable to mount Th2 responses during schistosomiasis. Although WT mice are protected from superinfection, resistance is abrogated in the absence of interleukin (IL)-4. We conclude that IL-4 or IL-4 dependent responses, or both, are necessary for resistance to S. mansoni superinfection in mice. PMID- 10461959 TI - Schistosoma japonicum is less sensitive to cyclosporin A in vivo than Schistosoma mansoni. AB - We compared the toxic effects of cyclosporin A (CsA) on postmigratory immature Schistosoma mansoni and Schistosoma japonicum in mice. For each species, CsA was administered either relatively early or late during development. Exposure of 20 day-old S. mansoni to 1 subcutaneous dose of CsA (50 mg/kg) reduced the worm burden by 45% and induced herniae and/or boli of the gut in 32% of perfusable worms. These results agree with previous reports. In addition, CsA induced a marked liver shift (37% of total worm number). For S. japonicum, CsA was administered at 11 days postinfection (PI) because this species migrates more quickly. Killing of worms and damage to the gut were not observed, and only a slight liver shift occurred. Similarly, these effects were not recorded when CsA was administered at the later times of 34 days PI for S. mansoni and 17 days PI for S. japonicum. For both species, CsA stunted worms, affecting both sexes early PI but only females late PI. In conclusion, immature worms of S. japonicum are less sensitive than S. mansoni to CsA. Also, S. mansoni displays marked age dependent differences in its sensitivity to CsA. PMID- 10461960 TI - Age-dependent infectivity of orally transferred juvenile Fasciola hepatica. AB - Juvenile Fasciola hepatica is infective when administered orally. To determine whether the age of juveniles is a factor in infectivity by oral transfer, experimental mice were challenged orally with immature F. hepatica that had been grown in donor mice for 12, 14, 16, and 18 days. Experimental mice were examined for infections 12 16 days after the oral transfers. The infection success in experimental mice decreased with the age of juveniles. The worm recovery also decreased according to the age of juveniles. None of the juveniles was infective when grown for longer than 11 days. Once infected, orally transferred worms continued to grow. Juvenile age was a significant factor in determining the infectivity of orally transferred juvenile F. hepatica. PMID- 10461961 TI - Secretion of multilamellar whorls by Eimeria tenella zoites. AB - Multilamellar whorls were demonstrated by transmission electron microscopy to be associated with sporozoites and all generations of merozoites of Eimeria tenella, in chicken cecal tissue fixed without tannic acid or ruthenium red at room temperature. Whorls were found within the parasitophorous vacuoles of recently invaded cells at all stages of development, suggesting a role in the formation of the host parasite interface. Whorls were also associated with intraluminal third generation merozoites prior to host cell invasion and appeared to be secreted directly through the pellicle. Membranous sheaths, shown by serial sectioning to be derived from intracellular whorl material, were observed enveloping some intraluminal merozoites. In many third-generation merozoites, whorl material was located within discrete novel organelles (here termed lamellosomes) located in the apical region. These densely staining spherical organelles were morphologically distinct from micronemes and rhoptries and were one-third the size of dense granules. These findings confirm that whorls are nonartifactual secretions whose lamellar organization is lost during normal fixation on ice without tannic acid. It is hypothesized that whorls secreted prior to invasion are involved in protection of the motile zoite, immune evasion, or some aspect of gliding motility. PMID- 10461962 TI - Congenital transmission of experimental chronic toxoplasmosis in rats. AB - A 10% transplacental transmission rate was observed in litters from 89 Wistar rats chronically infected with Toxoplasmosis gondii, as judging from bioassays. The rats had been fed T. gondii 2 mo prior to mating. Six of 7 isolates of T. gondii were transplacentally transmitted. The frequency of transmission did not appear to be affected by the strain of T. gondii or the size of the inoculum. PMID- 10461963 TI - Prevalence of Sarcocystis kirkpatricki Sarcocysts in the central nervous system and striated muscles of raccoons from the Eastern United States. AB - A retrospective survey of 760 raccoons (Procyon lotor) revealed 9 animals with sarcocysts of Sarcocystis kirkpatricki in their brains. Six of the raccoons also had the organisms in their skeletal muscles, and 1 had them in the heart muscle. No age or gender predisposition was seen. Seven of the raccoons had concurrent viral diseases (canine distemper or rabies), suggesting that concurrent viral infections in raccoons may facilitate infection of brain tissue with S. kirkpatricki. PMID- 10461964 TI - Comparison of the internal transcribed spacer, ITS-1, from Sarcocystis falcatula isolates and Sarcocystis neurona. AB - The genetic diversity among 6 Sarcocystis falcatula isolates derived from geographically distinct regions in the U.S.A. was detected using the first internal transcribed spacer region 1 (ITS-1) of the rRNA gene. These sequences were then compared to the full sequence from a Sarcocystis neurona isolate obtained from a California horse diagnosed with equine protozoal myeloencephalitis. No nucleotide differences were detected over partial sequence analysis of 2 additional S. neurona isolates: however, the complete nucleotide sequence for the ITS-1 region was not compared. Twelve nucleotide differences were consistently detected when aligned sequences of S. neurona were compared to those of the S. falcatula isolates. Additional nucleotide base changes were detected among the S. falcatula isolates, but these changes were not consistent in all the S. falcatula isolates. These results indicate that S. falcatula may be comprised of a heterogeneous population and that the ITS-1 region can be used to distinguish S. neurona from S. falcatala used in this study. PMID- 10461965 TI - Validation of a rapid method for extraction of total RNA applied to Leishmania promastigotes. AB - This report explains a rapid procedure (approximately 50 min) for the isolation of highly purified total RNA from Leishmania promastigotes based on extraction with acidic phenol. The simplicity of the manipulations required make this method ideal for processing multiple samples; the quality of the RNA obtained is suitable for reverse transcription polymerase chain reaction analysis. PMID- 10461966 TI - Trichinella pseudospiralis in sedentary night-birds of prey from Central Italy. AB - Trichinella pseudospiralis has been isolated from carnivorous and carrion-feeding mammals and birds in Eastern Europe, Asia, Australia, and North America, suggesting its cosmopolitan distribution. We conducted a survey to detect this parasite in raptorial and carrion-feeding birds in Italy, examining muscles from 205 animals by artificial digestion. We isolated from the breast muscle 1 larva from a tawny owl (Strix aluco) and 2 larvae from a little owl (Athene noctua). These larvae were identified as T. pseudospiralis by the polymerase chain reaction with a specific primer set. This is the first documented report of T. pseudospiralis in animals in Western Europe. PMID- 10461967 TI - Diet texture modifies outcome of a primary infection with Heligmosomoides polygyrus (nematoda) in mice. AB - To determine if dietary texture altered the establishment, survival and reproduction of Heligmosomoides polygyrus during a primary infection, commercial mouse chow (Purina Chow) was fed in either pelleted or powdered form to 2 groups of infected BALB/c mice, and a semipurified, biotin-fortified, egg-white-based diet was provided in powdered form to a third group of mice. Diet texture (powdered vs. pelleted) modified parasite establishment, as evidenced by higher worm recovery 6 days postinfection (PI) in both groups of mice fed powdered diets compared with the group fed the pelleted diet, but diet texture had no detectable effect on net or per capita egg production or on day 30 worm survival. However, almost twice as many worms were recovered from mice fed the semipurified, powdered diet than those fed either textural formulation of the commercial mouse chow on day 30 PI, indicating that a dietary component in the semipurified diet, such as the single fiber source cellulose, facilitated parasite survival, or that other ingredients in the commercial mouse chow, perhaps the more complex fiber components, reduced worm survival, or both. We conclude that dietary texture influences parasite establishment and that specific fibers that change texture can individually and independently modify H. polygyrus survival. PMID- 10461968 TI - Accumulation of mucosal T lymphocytes around epithelial cells after in vitro infection with Cryptosporidium parvum. AB - We had previously shown that ileal intraepithelial lymphocytes isolated from calves with cryptosporidiosis include significantly increased numbers of CD8+ T lymphocytes and activated CD4+ cells. These increases could result from redistribution of resident mucosal lymphocytes or from homing of peripheral T cells to ileal mucosa. To determine whether resident mucosal lymphocytes can redistribute to Cryptosporidium parvum-infected epithelium, oocysts were inoculated in vitro onto ileum explants taken from 1-2-wk-old noninfected calves. After 24 hr of incubation, the explants were collected and frozen in liquid nitrogen. Immunohistochemical analysis of T-lymphocyte subpopulations was performed on sections, and labeled lymphocytes adjacent to villous epithelial cells were counted. Compared with uninoculated explants, there was a statistically significant increase in the number of CD8+ T lymphocytes per 100 epithelial cells in oocyst-inoculated tissue. In addition, there were increased numbers of CD4+ T cells and activated (CD25+) lymphocytes adjacent to C. parvum infected epithelium. These results show that resident mucosal T lymphocytes can accumulate at the epithelium during C. parvum infection. PMID- 10461969 TI - Revisiting the code: clarifying name-bearing types for photomicrographs of protozoa. PMID- 10461970 TI - Effect of prepubertal feeding regimen on reproductive development of gilts. AB - The effect of prepubertal feed level on growth and reproductive development of gilts was investigated. At 13 wk. of age, white crossbred gilts were penned individually and assigned to the following treatments: Ad lib, ad libitum intake from 13 to 25 wk. of age (n = 64); Control, ad libitum intake from 13 wk. of age until 100 kg BW and then 90% of ad libitum intake until 25 wk. of age (n = 65); and Restricted, 74% of ad libitum intake from 13 wk. to 25 wk. of age (n = 64). Feed was formulated to primarily restrict energy intake. The study was replicated in two seasons. At 25 wk. of age, gilts were moved to group pens, approximately 16 gilts/pen, allowed ad libitum access to feed, and estrus detection was initiated. Gilts were mated at first estrus and those recycling were remated. After mating, gilts were moved to gestation stalls and fed 1.5x maintenance. At 30 d of gestation, reproductive tracts were harvested, and numbers of corpora lutea (CL) and live embryos were recorded. From 13 to 25 wk. of age, feed consumption was 258 for Ad lib, 251 for Control, and 189 kg/gilt for Restricted, and, from 13 wk. of age until 30 d of gestation, total feed consumption was 367 for Ad lib, 356 for Control, and 299 kg/gilt for Restricted gilts. Age at puberty (196 d) and pregnancy (200 d) was not affected (P>.18) by treatment. However, the rate at which gilts attained puberty (e.g., percentage pubertal at 28 d) was greatest in Ad lib (75) and least in Control (61) gilts. Number of CL and live embryos at 30 d of gestation/gilt assigned to the study was unaffected (P>.21) by treatment. Quantity of feed consumed from 13 wk. of age to 30 d of gestation per live embryo in gilts assigned to the study was 40.0 for Ad lib, 39.8 for Control, and 30.6 kg/gilt for Restricted gilts. These results indicate that moderate feed restriction of gilts during prepubertal development may increase efficiency of swine production without negative impact on reproductive performance through 30 d of gestation. PMID- 10461971 TI - Using current on-line carcass evaluation parameters to estimate boneless and bone in pork carcass yield as influenced by trim level. AB - The objective of this study was to develop prediction equations for estimating proportional carcass yield to a variety of external trim levels and bone-in and boneless pork primal cuts. Two hundred pork carcasses were selected from six U.S. pork processing plants and represented USDA carcass grades (25% USDA #1, 36% USDA #2, 25% USDA #3, and 14% USDA #4). Carcasses were measured (prerigor and after a 24 h chill) for fat and muscle depth at the last rib (LR) and between the third and fourth from last rib (TH) with a Hennessy optical grading probe (OGP). Carcasses were shipped to Texas A&M University, where one was randomly assigned for fabrication. Selected sides were fabricated to four lean cuts (ham, loin, Boston butt, and picnic shoulder) then fabricated progressively into bone-in (BI) and boneless (BL) four lean cuts (FLC) trimmed to .64, .32, and 0 cm of s.c. fat, and BL 0 cm trim, seam fat removed, four lean cuts (BLS-OFLC). Total dissected carcass lean was used to calculate the percentage of total carcass lean (PLEAN). Lean tissue subsamples were collected for chemical fat-free analysis and percentage carcass fat-free lean (FFLEAN) was determined. Longissimus muscle area and fat depth also were collected at the 10th and 11th rib interface during fabrication. Regression equations were developed from linear carcass and OGP measurements predicting FLC of each fabrication point. Loin muscle and fat depths from the OPG obtained on warm, prerigor carcasses at the TH interface were more accurate predictors of fabrication end points than warm carcass probe depth obtained at the last rib or either of the chilled carcass probe sites (probed at TH or LR). Fat and loin muscle depth obtained via OGP explained 46.7, 52.6, and 57.1% (residual mean square error [RMSE] = 3.30, 3.19, and 3.04%) of the variation in the percentage of BI-FLC trimmed to .64, .32, and 0 cm of s.c. fat, respectively, and 49.0, 53.9, and 60.7% (RMSE = 2.91, 2.81, and 2.69%) of the variation in the percentage of BL-FLC trimmed to .64, .32, and 0 cm of s.c. fat, respectively. Fat and loin muscle depth from warm carcass OGP probes at the TH interface accounted for 62.4 and 63.5% (RMSE = 3.38 and 3.27%) of the variation in PLEAN and FFLEAN, respectively. These equations provide an opportunity to estimate pork carcass yield for a variety of procurement end point equations using existing on-line techniques. PMID- 10461972 TI - Effect of level of chronic immune system activation on the lactational performance of sows. AB - The effect of the level of chronic immune system (IS) activation on sow lactational performance was determined in 11 pairs of littermate, primiparous sows. Sows with a low level of IS activation were created by rearing the animals via early weaning, isolated rearing schemes. During lactation, two levels of IS activation were achieved in each littermate sow pair by subcutaneous administration of either 0 (saline) or 5 microg/kg of sow BW of Escherichia coli lipopolysaccharide (LPS) in a mineral oil adjuvant emulsion on d 2 and 10 of lactation. Litters were standardized to 13 pigs by 8 h postpartum. Sows were offered daily 6.0 kg of a corn-soy diet formulated to contain a minimum of 250% of the dietary nutrient concentrations estimated to be needed by lactating sows. Based on antibody titers to LPS and serum concentrations of alpha-1 acid glycoprotein (AGP), high IS sows mounted an immune response to the LPS during lactation, and low IS sows maintained a low level of IS activation. Over an 18-d lactation, a high level of chronic activation of the sows' immune systems depressed daily sow feed intakes by .56 kg, litter weight gains by .32 kg, and daily milk by 1.4 kg, milk energy by 1.7 Mcal, and milk protein yields by 71 g, but did not alter sow body weight loss. The reductions in yields of milk and milk nutrients likely were because of proinflammatory cytokine-induced inhibition of the lactogenic hormones resulting from high chronic IS activation. Based on these data, the level of chronic IS activation alters the lactational performance of sows. PMID- 10461973 TI - Threshold-linear versus linear-linear analysis of birth weight and calving ease using an animal model: I. Variance component estimation. AB - Birth weight and calving difficulty were analyzed with Bayesian methodology using univariate linear models, a bivariate linear model, a threshold model for calving difficulty, and a joint threshold-linear model using a probit approach. Field data included 26,006 records of Gelbvieh cattle. Simulated populations were generated using parameters estimated from the field data. The Gibbs sampler was used to obtain estimates of the marginal posterior mean and standard deviation of the (co)variance components, heritabilities, and correlations. In the univariate analyses, the posterior mean of direct heritability for calving difficulty was .23 with the threshold model and .18 with the linear model. Maternal heritabilities were .10 and .08, respectively. In the bivariate analysis, posterior means of direct heritability for calving difficulty were .21 and .18 for the bivariate linear-threshold and linear-linear model, respectively. Maternal heritabilities were .09 and .06, respectively. Direct heritability for birth weight was .25 for the univariate model and .26 for bivariate models. Maternal heritability was .05 for the linear-threshold model and the univariate model and .06 for the bivariate linear model. Genetic correlation between direct genetic effects in both traits was .81 for the linear-threshold model and .79 for the bivariate linear. Residual correlation was .35 for the bivariate linear model and .50 for the bivariate linear-threshold. A simulation study confirmed that the posterior mean of the marginal distribution was suitable as a point estimate for univariate threshold and bivariate linear-threshold models. PMID- 10461974 TI - Threshold-linear versus linear-linear analysis of birth weight and calving ease using an animal model: II. Comparison of models. AB - Several models were evaluated in terms of predictive ability for calving difficulty. Data included birth weight and calving difficulty scores provided by the American Gelbvieh Association from 26,006 calves born to first-parity cows and five simulated populations of 6,200 animals each. Included in the model were fixed age of dam x sex interaction effects, random herd-year-season effects, and random animal direct and maternal effects. Bivariate linear-threshold and linear linear models for birth weight/calving ease and univariate threshold and linear models for calving ease were applied to the data sets. For each data set and model, one-half of calving ease records were randomly discarded. Predictive ability of the different models was defined with the mean square error (MSE) for the difference between a deleted calving ease score and its prediction obtained from the remaining data. In terms of correlation between simulated and predicted breeding values, the threshold models had a 1% advantage for direct genetic effects and 3% for maternal genetic effects. In simulation, the average MSE was .29 for linear-threshold, .32 for linear-linear, .37 for threshold, and .39 for linear model. For the field data set, the MSE was .31, .33, .39, and .40, respectively. Although the bivariate models for calving ease/birth weight were more accurate than univariate models, the threshold models showed a greater advantage under the bivariate model. For the purpose of genetic evaluation for calving difficulty in beef cattle, the use of the linear-threshold model seems justified. In dairy cattle, the evaluation for calving ease can benefit from recording birth weight. PMID- 10461975 TI - Two-stage selection strategies utilizing marker-quantitative trait locus information and individual performance. AB - Short- and long-term response to marker-assisted selection in two stages was studied using a stochastic simulation of a closed nucleus herd for beef production. First-stage selection was carried out within families based on information at a fully additive quantitative trait locus (QTL). Second-stage selection strategies were based on 1) individual phenotype, 2) individual phenotype precorrected for QTL, 3) a selection index incorporating phenotype and QTL information, 4) a standard animal model BLUP, and 5) a selection index incorporating marker-QTL information and standard animal model BLUP on records precorrected for QTL. All strategies were efficient in moving the favorable allele at the QTL to fixation, but they differed in the time to reach fixation. Mass selection was less efficient in changing allele frequencies than BLUP. Discounted accumulated response, accounting for the time response was realized and inflation rate, was proposed to rank strategies and to elude the conflict between short- and long-term response in marker-assisted selection. Discounted accumulated response at a time horizon of 20 yr for alternative two-stage selection strategies was compared with conventional BLUP carried out in second stage only. Within-family selection increased discounted accumulated response by more than 11% using Strategy 4 and by up to 12% using Strategy 5 at an inflation rate of 2%. The percentage increase in response was less for highly heritable traits and when the proportion of additive variance explained by the QTL was small. Strategy 5 gave larger response with reduced inbreeding. This strategy also resulted in the lowest cost-benefit ratio, requiring less genotyping per unit of response. Cost-benefit ratio for discounted genotyping and for discounted in vitro production of embryos for traits with low heritability was two to four times that for traits with high heritability. The use of first-stage selection slightly increased the level of inbreeding for both mass (Strategy 1) and BLUP selection (Strategies 4 and 5). PMID- 10461976 TI - Genetic trends and breed overlap derived from multiple-breed genetic evaluations of beef cattle for growth traits. AB - Genetic evaluations for a multiple-breed population of beef cattle were used to estimate genetic trends for five breeds, and genetic differences and overlap among 14 breeds. Genetic evaluations studied were for direct contributions to birth weight, gain from birth to 200 and 365 d, and maternal contribution to gain from birth to 200 d. Almost all genetic trends were positive, but the magnitude of the trends varied among breeds. Trends were nonlinear between 1985 and 1995 for most breed and trait combinations. The rates of increase in genetic trends were generally higher for the lighter weight breeds, and lighter weight breeds had faster growth rate genetic trends at 1995 than the heavier breeds. Genetic trend estimates for yearling gain at 1995 were 2.46, 2.23, 1.73, 1.70, and 1.46 kg/yr for Angus, Hereford, Limousin, Charolais, and Simmental, respectively. Corresponding birth weight genetic trends were .130, .226, .049, .130, and .048 kg/yr. Mean genetic differences between breeds have been decreasing in magnitude due to these differences in genetic trends between heavier and lighter breeds. Genetic variation for the traits studied seemed to be greater within than between breeds for calves born and cows calving between 1993 and 1995. Genetic trends at 1995 suggest that ratios of within:between breed variation will increase and that across-breed genetic improvement initiatives for growth traits will become more important in the future. PMID- 10461977 TI - Technical note: direct genotyping of the double-muscling locus (mh) in Piedmontese and Belgian Blue cattle by fluorescent PCR. AB - A simple PCR-based allele detection system has been developed to assist in the management of the two most prevalent double-muscled (mh) breeds in the U.S. Application of this assay will permit the implementation of structured mating systems dependent on precise genotypes at the mh locus. The genetic assay uses standard fluorescent genotyping technology and relies on the unique nucleotide composition of wild-type and mutant alleles of myostatin, the gene underlying the double-muscled phenotype. We present data demonstrating the efficacy of this fluorescent primer-based PCR assay in genotyping animal populations carrying normal and(or) mutant alleles of the myostatin gene. PMID- 10461978 TI - Shrinking membership in the American Society of Animal Science: does the discipline of poultry science give us some clues? AB - Concerns have been expressed by the American Society of Animal Science (ASAS) leadership about the declining membership in ASAS. I present the viewpoint that the history of the Poultry Science Association (PSA) membership and the elimination of poultry science departments from many land grant universities could be an indication of what the future holds for animal science. I suggest that the industrialization of poultry production has led to a decline in the demand for traditionally trained poultry scientists. Industrialization of swine production is proceeding rapidly, with other animal-based industries showing the same trend. If maintaining a large ASAS membership is indeed a priority, new opportunities must be developed. Equine and companion animal programs offer such possibilities, tapping into a high level of student interest. PMID- 10461979 TI - Effect of pen size on behavioral, endocrine, and immune responses of water buffalo (Bubalus bubalis) calves. AB - Female water buffalo (Bubalus bubalis) calves (n = 28) aged 7 to 10 d were divided into four groups of seven animals each to examine the effects of space allowance (Group A: 2.6 indoor m2 + 2.0 outdoor m2/calf; Group B: 2.6 indoor m2/calf; Group C: 1.5 indoor m2/calf; Group D: 1.0 indoor m2/calf) on behavioral, endocrine, and immune variables for a period of 60 d. Animals were offered 7 L/d of a commercial acidified milk substitute. The calves averaged 45.9 kg initially and 92.4 kg finally. The behavior observations were conducted 7 d after grouping and fortnightly thereafter. At wk 4 and 8, the phytohemagglutinin (PHA) skin test was performed to induce aspecific delayed hypersensitivity. At wk. 1 and 3, calves were injected i.m. with keyhole limpet hemocyanin. Antibody titers were determined at weekly intervals for 7 wk. Calves in pens with greater space allowance (Groups A and B) were less active than Groups C and D (P<.001). The latter groups were also observed feeding more often at wk 7 (P<.01). Calves provided with an outdoor paddock spent less time standing than Groups C and D (P<.01), and lay with a greater number of outstretched legs (P<.001). Groups C and D showed a lower reaction to PHA in both skin tests than did Groups A and B (P<.001 and P<.05, respectively). Group A showed an antibody response consistently higher than groups B, C, and D (P<.01, P<.05, and P<.05, respectively). At the end of the experimental period, the calves were subjected to an isolation test lasting 10 min. Group D showed a longer duration of movement with respect to Groups A and B (P<.01); animals from Group C walked more than did Group A (P<.05). Cortisol concentration evaluated 0, 10, 45, 90, 150, and 225 min after separation from the group was higher in Groups C and D than in Groups A and B (P<.01). For all animals, the highest cortisol level was observed immediately after the isolation test (P<.001). Space restriction resulted in evidence of stress in the animals as shown by alterations in a number of physiological responses. However, the use of small groups of only seven animals per pen may have affected their reactions to space restriction. It is possible that using larger groups could change these conclusions. PMID- 10461980 TI - Adrenocorticotropic hormone and cortisol in calves after corticotropin-releasing hormone. AB - The aim for this study was to analyze responsiveness of the hypothalamo-pituitary adrenocortical axis to exogenous bovine corticotropin-releasing hormone (bCRH) in calves. Two dose-response studies were carried out, using either bCRH alone (dose rates of 0, .01, .03, and .1 microg bCRH/kg live weight) or in combination with arginine-vasopressin (bCRH:AVP, 0:0, .1:.05, .5:.25, and 1:.5 microg kg live weight). The bCRH was administered i.v. to calves (n = 5 to 7 per dose) housed individually or in groups. Serial blood samples were obtained from before to 300 min after injection and analyzed for plasma ACTH and cortisol concentrations. The lowest bCRH dose that produced a response in all calves was .1 microg/kg. In the experiment using bCRH with AVP, increasing the bCRH dose from .1 to 1 microg/kg resulted in an increase in peak ACTH concentration (321 vs. 2,003 pg/mL) but did not significantly affect the peak cortisol concentration (37 vs. 40 ng/mL). The time to reach the peak cortisol concentration increased with the dose of bCRH with AVP (from 38 to 111 min). The ACTH and cortisol concentrations determined at any time between 20 and 90 min after bCRH injection were correlated to the integrated responses calculated as areas under the ACTH and the cortisol curves (r between .61 and .99, P<.05). In comparison with results from studies in humans, pigs, and sheep, our data showed that the pituitary of calves seems less sensitive to CRH than that of other mammals, despite a greater capacity to produce ACTH. Moreover, the calf's adrenals seem to have a lower capacity to produce cortisol than adrenals of other mammals. As in other species, it seems that AVP enhances the release of ACTH and cortisol. For CRH challenge to be used in calves, we suggest injecting at least .1 microg of bCRH/kg live weight either with or without AVP and taking several blood samples before injection and between 20 and 90 min after injection. PMID- 10461981 TI - Butorphanol tartrate acts to decrease sow activity, which could lead to reduced pig crushing. AB - The objective of this study was to determine whether administration of an analgesic to sows immediately after farrowing would allow them to lie more restfully. Sows lying on their pigs, causing them to be "crushed," is a major cause of pig mortality. Most deaths due to crushing occur during the first 3 d postpartum. For modern, lean-type sows, farrowing crates are relatively hard and unforgiving, even though they may be equipped with plastic-coated, expanded metal flooring. Indeed, many sows develop pressure sores on their shoulders, and this may contribute to the sows' discomfort. These sores may cause a sow to change position frequently to alleviate pain, thus increasing its chances of crushing pigs. Sixteen production sows were assigned to either a control group (C, n = 8) with litter size 11.71+/-.78 or an experimental group (B, n = 8) with litter size 11.63+/-1.22. Pigs born to C and B sows weighed 1.60+/-.04 and 1.37+/-.04 kg, respectively. The C sows were given no treatment, whereas the B sows were administered an i.m. injection of butorphanol tartrate at a dose of .15 mg/kg BW every 6 h until 3 d after farrowing. Data were collected on all sows using time lapse photography (1 frame/.4 s) for a 3-d duration upon the initiation of farrowing. To assess the degree of comfort of each sow, body position changes were recorded when sows switched between lying, sitting, and standing. Data were analyzed by 12-h periods using Wilcoxon-Mann-Whitney equations. During the 72-h period, B sows tended to perform fewer position changes than C sows (P = .10). Specifically, position changes were fewer for B sows from 48 to 72 h postpartum (P<.06). There were no differences in position changes between treatments from 0 to 48 h postpartum (P>.50). There was no difference in the rate of crushing between treatments (C = 5, B = 5). The butorphanol did not seem to affect pig activity or normal behaviors or to create adverse effects, such as diarrhea. Although the sows given butorphanol showed a reduced number of position changes, the dose was intermediate, and a higher dose may have a greater effect. If pig mortality can be decreased, an analgesic protocol that is simple to administer and readily available to producers can be developed. Alternatively, using of more pliable flooring or an increase in sow body fat may allow sows to lie more stationary. PMID- 10461982 TI - Danger to pigs due to crushing can be reduced by the use of a simulated udder. AB - Sows that lie on their young, pig "crushing", is a significant cause of pig mortality in current production systems. Although mortality rates of pigs in farrowing crates are lower than mortality rates of pigs in pens, loss due to crushing is still estimated to be between 4.8 and 18%. During the first few days after parturition, pigs are highly attracted to the odor of their dam's udder. Thus, our research was designed to move the pigs away from the sow by competing with the sow's udder using a "simulated" udder. Fifteen Yorkshire x Landrace sows and their litters (11.4+/-.78 pigs) were assigned to either a control (C, n = 9) or an experimental group (SU, n = 6). The C pigs had access to a heat lamp, whereas the SU pigs' crate had a simulated udder. Data were collected using time lapse photography (1 frame/.4 s) for a 3-d duration at the initiation of farrowing. When a sow stood, data were recorded by 1-min scan samples to record the number of pigs using either the heat lamp or the simulated udder. In addition, stillborn pigs, pig crushing, and death by other means also were recorded. Data were analyzed by 12-h periods using generalized estimating equations. Results indicate that from 12 to 72 h postpartum, excluding 24 to 36 h postpartum, the estimated probability that pigs were in a safe area (simulated udder or heat lamp) was .89 for SU pigs, compared with only .72 for C pigs (P = .005). During the 24- to 36-h period, it was more probable to find pigs on a simulated udder (.77) than under only a heat lamp (.61, P = .016). Stillborn pigs, pig crushing, and death by other means were not different between treatments (mean = .87, .60, 1.2; P>.20). The simulated udder drew pigs away from the sow's udder better than heat lamps alone. Considering these findings, mortality of pigs due to crushing may be decreased substantially using a simulated udder. These results are promising, but further refinement should be done, including improved udder design and investigation of the attractiveness of various stimuli. PMID- 10461983 TI - Shade and wind barrier effects on summertime feedlot cattle performance. AB - In each of three summertime trials conducted over consecutive years, approximately 110 predominantly black and black-white-face steers were blocked by weight and randomly allotted to one of 16 pens in a 2x2 factorial arrangement of treatments. Factors consisted of cattle being fed in facilities with or without wind barriers and with or without shade. Steers were fed dry-rolled corn-based diets (1.43 Mcal/ kg, NEg). Mean starting date and days on feed were June 26 and 79, respectively. In unshaded areas, temperature and humidity averaged 21.6 degrees C and 77.9%, and the blackglobe-humidity index (BGHI) at 1500 averaged between 84.0 and 89.1. Each of four 6.1-x6.1-m structures (mean height = 3.4 m) with white steel roofs provided shade (2.65 m2/steer) for two pens. In facilities with wind barriers provided, airflow was reduced from the north and northwest by a 25-m-wide shelterbelt containing six rows of trees. For cattle fed in pens with wind barriers, shade increased (P<.05) gain from 0 to 56 d and decreased (P<.05) DMI/ADG from 0 to 28 d. Differences (P<.05) in performance were not found between shaded and unshaded cattle in any portion of the feeding period for cattle fed in the pens without wind barriers and over the entire feeding period in either type of facility. The shade response in pens with wind barriers seemed to be greater the 1st yr than in subsequent years. Differences in weather patterns among years, especially air temperature, humidity, and solar radiation, may partially explain this interaction. Also, in yr 1, cattle tended to have greater fat thickness at finish than in yr 2 and 3. Correlations between BGHI and DMI tended to be greater during the early portion of the trial (0 to 28 d) than over the entire trial. Correlations between the difference in BGHI under shade vs no shade and percentage of shade use had the greatest magnitude and were significant only in the first 28 d vs over the entire feeding period. Although no heat-related cattle deaths occurred in this study, results suggest that shade improves cattle performance in the summer when they are fed in facilities with winter wind protection available and have not become acclimated to hot conditions. Once cattle are acclimated or hot conditions subside, compensation by unshaded cattle offsets much of the initial benefits of providing shade. PMID- 10461984 TI - Facilitation of sexual behavior in French-Alpine goats treated with intravaginal progesterone-releasing devices and estradiol during the breeding and nonbreeding seasons. AB - The effectiveness of administering progesterone (P4) using controlled intravaginal drug release (CIDR) devices on estradiol (E2)-induced sexual behaviors was examined in ovariectomized (ovx) French-Alpine goats during the fall and spring. Estradiol-induced attractivity and receptivity were facilitated during the spring when P4-filled CIDR devices were removed 24 or 48 h before injection of 30 microg of E2. During the fall, attractivity was also facilitated by CIDR removal 24 h prior to E2 injection, whereas E2-induced receptivity was unaffected by removal of the CIDR at this interval. Concentrations of P4 in circulation during the 3 d of treatment with a CIDR were similar to those during the late luteal phase of the estrous cycle in intact goats. Treatment with P4 filled CIDR for 3 d, followed by injection with 30 microg of E2 24 h after removal, was determined to be a useful model for inducing sexual behavior in a physiologically relevant manner, and it may also be an effective means for facilitating estrus detection due to the high frequency of display of sexual behavior during a predictable time period following steroid treatment. PMID- 10461985 TI - Effects of oral chlortetracycline and dietary protein level on plasma concentrations of growth hormone and thyroid hormones in beef steers before and after challenge with a combination of thyrotropin-releasing hormone and growth hormone-releasing hormone. AB - The objective of this study was to determine the effect of a subtherapeutic level of chlortetracycline (CTC) fed to growing beef steers under conditions of limited and adequate dietary protein on plasma concentrations of GH, thyroid-stimulating hormone (TSH), and thyroid hormones before and after an injection of thyrotropin releasing hormone (TRH) + GHRH. Young beef steers (n = 32; average BW = 285 kg) were assigned to a 2x2 factorial arrangement of treatments of either a 10 or 13% crude protein diet (70% concentrate, 15% wheat straw, and 15% cottonseed hulls) and either a corn meal carrier or carrier + 350 mg of CTC daily top dressed on the diet. Steers were fed ad libitum amounts of diet for 56 d, and a jugular catheter was then placed in each steer in four groups (two steers from each treatment combination per group) during four consecutive days (one group per day). Each steer was injected via the jugular catheter with 1.0 microg/kg BW TRH + .1 microg/kg BW GHRH in 10 mL of saline at 0800. Blood samples were collected at -30, -15, 0, 5, 10, 15, 20, 30, 45, 60, 120, 240, and 360 min after releasing hormone injection. Plasma samples were analyzed for GH, TSH, thyroxine (T4), and triiodothyronine (T3). After 84 d on trial, the steers were slaughtered and the pituitary and samples of liver were collected and analyzed for 5'-deiodinase activity. Feeding CTC attenuated the GH response to releasing hormone challenge by 26% for both area under the response curve (P<.03) and peak response (P<.10). Likewise, CTC attenuated the TSH response to releasing hormone challenge for area under the response curve by 16% (P<.10) and peak response by 33% (P<.02), and attenuated the T4 response for area under the curve by 12% (P<.08) and peak response by 14% (P<.04). Type II deiodinase activity in the pituitary was 36% less (P<.02) in CTC-fed steers than in steers not fed CTC. The results of this study are interpreted to suggest that feeding subtherapeutic levels of CTC to young growing beef cattle attenuates the release of GH and TSH in response to pituitary releasing hormones, suggesting a mechanism by which CTC may influence tissue deposition in cattle. PMID- 10461986 TI - The interaction of hydrocortisone and thyroxine during fetal adipose tissue differentiation: CCAAT enhancing binding protein expression and capillary cytodifferentiation. AB - Late-term fetal pigs from genetically obese dams have elevated levels of thyroid hormones and glucocorticoids, depressed levels of GH, larger fat cells and elevated lipogenesis than do fetal pigs from lean dams. We investigated the influence of elevated levels of thyroid hormones and glucocorticoids per se on adipose tissue traits by chronically treating hypophysectomized (hypox; d 70) fetal pigs between d 90 and 105 of gestation with either thyroxine (T4), hydrocortisone (HC), or the combination of T4 + HC. Treatment with T4 and T4 + HC increased serum T4 and IGF-I levels and enhanced skin and hair development. Treatment with HC and T4 + HC increased serum HC levels, fat cell size, and inner subcutaneous adipose tissue thickness. Quantitative analysis of stained adipose tissue sections indicated that T4 + HC treatment increased lipid accretion and fat cell cluster development more than did either hormone alone. The T4 + HC markedly increased apparent fat cell number, because there was only a 19% increase in fat cell size. A hypox-induced deficit in cytodifferentiation of capillaries associated with adipocytes was not influenced by T4, but was partially normalized by treatment with HC and T4 + HC. Immunocytochemical and Western blot analyses showed no influence of hormonal treatment on expression of three CCAAT enhancing binding protein (C/EBP) isoforms. However, expression of C/EBPdelta in adipose tissue was markedly reduced in control fetal pigs compared with hypox fetal pigs. These studies indicate that concurrent action of glucocorticoids and thyroid hormones may be the critical aspect of endocrine regulation of fetal adipogenesis. PMID- 10461987 TI - The relationship between endogenous insulin-like growth factors and growth in pigs. AB - Previous studies have reported conflicting data on gender differences in plasma IGF-I in postnatal pigs. There is also debate over the role of IGF-II in regulation of postnatal growth. We have, therefore, determined the concentrations of plasma IGF-I, IGF-II, and IGF-binding protein-3 (IGFBP-3) in boars, barrows, and gilts and related these to postnatal growth characteristics. Plasma concentrations of IGF-I were higher in boars than in gilts or barrows from 13 wk. of age, and plasma IGF-II levels were generally higher in barrows than in boars or gilts. Plasma IGFBP-3 levels were higher in boars than in gilts or barrows at most ages. Between 15 and 23 wk. of age, IGF-I and IGFBP-3, but not IGF-II, were positively associated with growth rate, voluntary feed intake, and gain:feed ratio. Plasma IGF-II, but not IGF-I or IGFBP-3, was positively associated with backfat depth during this period. These results support the hypothesis that circulating IGF-I and IGF-II are regulators of lean and adipose tissue growth, respectively. PMID- 10461988 TI - Correlation between histochemically assessed fiber type distribution and isomyosin and myosin heavy chain content in porcine skeletal muscles. AB - Highly sensitive enzyme assays developed to differentiate skeletal muscle fibers allow the recognition of three main fiber types: slow-twitch oxidative (SO), fast twitch oxidative glycolytic (FOG), and fast-twitch glycolytic (FG). Myosin, the predominant contractile protein in mammalian skeletal muscle, can be separated based on the electrophoretic mobility under nondissociating conditions into SM2, SM1, IM, FM3, and FM2 isoforms, or under dissociating conditions into myosin heavy chain (MHC) I, IIb, IIx/d, and IIa. The purpose of the present study was to determine whether the histochemical method of differentiation of fiber types is consistent with the electrophoretically identified isomyosin and MHC isoforms. These comparisons were made using serratus ventralis (SV), gluteus medius (GM), and longissimus muscles (LM) from 13 pigs. Two calculation methods for the histochemical assessed fiber type distribution were adopted. The first method incorporated the number of fibers counted for each fiber type and calculated a percentage of the total fiber number (fiber number percentage: FNP). The second method expressed the cross-sectional area of each fiber type as a percentage of the total fiber area measured per muscle (fiber area percentage: FAP). Independent of the calculation methods, correlation analyses revealed in all muscles a strong relation between SO fibers, the slow isomyosin (SM1 and SM2), and MHCI, as well as between the FG fibers, the fast isomyosin (FM3 and FM2), and MHCIIx/b content (P<.05). There were no correlations between FOG fiber population assessed by histochemical analysis and intermediate isoform (IM) or MHCIIa content. The present results did not provide conclusive evidence as to which of the calculation methods (FNP or FAP) was more closely related to myosin composition of skeletal muscles. Despite some incompatibility between the methods, the present study shows that histochemical as well as electrophoretic analyses yielded important information about the composition of porcine skeletal muscle. The combination of the two methods may be essential to accurately characterize porcine skeletal muscles. PMID- 10461989 TI - Identification of quality management practices to reduce the incidence of retail beef tenderness problems: development and evaluation of a prototype quality system to produce tender beef. AB - A prototype quality system for ensuring beef tenderness was designed and tested. The test population of cattle was genetically diverse, but it was constrained to include youthful (14- to 17-mo-old) steers with no more than 3/8 Bos indicus inheritance. Feeding and preharvest management of the cattle were consistent with procedures recommended for production of grain-finished beef of an acceptable quality level. In addition, the target endpoint for harvest (11-mm external fat thickness over the longissimus at the 12th rib) resulted in production of mostly Select and low Choice beef carcasses; 92% of the resulting carcasses qualified for these two grade levels. Application of the prototype quality system reduced the expected rate of nonconformance to desired tenderness specifications from about one in four loin steaks (23% for top sirloins and 26% for strip loins) to approximately one in eight loin steaks (13% for top sirloins and 12% for strip loins). Tenderness comparisons among sires suggested that the rate of nonconformance for strip loin steaks might be reduced even further by control of genetic inputs into the system. Use of process control in a quality management system was demonstrated to be an effective approach for assurance of beef tenderness. PMID- 10461990 TI - Impact of the hydrodyne process on tenderness, microbial load, and sensory characteristics of pork longissimus muscle. AB - Paired, boneless pork loin muscles were obtained from 76 market hogs to evaluate tenderness, meat quality characteristics, sensory attributes, and microbial characterization of pork muscle exposed to the Hydrodyne Process (H) compared with untreated control (C) loin. A subset of 16 paired loins was randomly selected for use in sensory evaluation and microbial characterization. Loins were vacuum packaged and immersed in a heat shrink tank prior to the H treatment. The Hydrodyne treatment exposed the loin to the pressure equivalent of a 150-g explosive, generating a pressure distribution of approximately 703 kg/cm2 at the surface of the samples. Meat quality assessments taken following treatment included subjective color, firmness/wetness, marbling scores (1 to 5 scale), Minolta reflectance and color readings, drip loss, and lipid content. The P-value for statistical significance for main effects and interactions was set at <.05 in all analyses. Administration of H resulted in a 17% improvement in Warner Bratzler shear force (2.69 vs. 3.24 kg), with the shear force similar at two end point cooking times (11 and 16 min) corresponding to approximately 75 and 83 degrees C, respectively. No differences between H and C were observed for color score, firmness score, Minolta L, Minolta Y, or drip loss on uncooked samples. The H loins had lower marbling scores (P<.05) and intramuscular lipid (P<.05) content than the paired C loin. Sensory evaluation on the randomly selected (n = 16) paired loins samples showed no improvement in Warner-Bratzler shear force. Sensory panelists were also unable to detect a difference between H and C loins for both initial and sustained tenderness scores. No differences between H and C loins were found for pork flavor, off-flavor, cohesiveness, or number of chews before swallowing, but H loins had a significantly lower juiciness score and more cooking loss than C loins. Microbial analysis results showed no differences in coliform bacteria counts, aerobic plate counts, and no detectable levels of Escherichia coli bacteria in any loins. The findings support the ability of the Hydrodyne procedure to improve tenderness without impacting other muscle quality attributes of pork. PMID- 10461991 TI - Influence of high ambient temperatures on performance of multiparous lactating sows. AB - Multiparous Large White sows (n = 63) were used to investigate the effects of five ambient temperatures (18, 22, 25, 27, and 29 degrees C) and two dietary protein contents on their lactation performance. At each temperature treatment, ambient temperature was maintained constant over the 21-d lactation period. Dietary protein content was either 14 or 17% with essential amino acids levels calculated not to be limiting. The animals had ad libitum access to feed between the seventh and the 19th day of lactation. Diet composition did not influence lactation performance. Over the 21-d lactation, feed intake decreased from 5.67 to 3.08 kg/d between 18 and 29 degrees C. Between d 7 and 19, the corresponding values were 7.16 and 3.48 kg/d, respectively. This decrease was curvilinear; an equation to predict voluntary feed intake (VFI) from temperature (T, degrees C) and body weight (BW, kg) is proposed: VFI = -49,052 + 1,213 T - 31.5 T2 + 330 BW .61 BW2 (residual standard deviation: 1,018). Skin temperature increased regularly with increased ambient temperature (34.6 to 37.4 degrees C between 18 and 29 degrees C), whereas udder temperature reached a plateau at 25 degrees C (38.3 degrees C). The gradient of temperature between skin and rectum was minimal (2 degrees C ) at 27 degrees C and remained constant at 29 degrees C. This constancy coincides with the marked reduction of feed intake. The respiratory rate increased from 26 to 124 breaths/min between 18 and 29 degrees C, and this indicates that the evaporative critical temperature was below 22 degrees C. The BW loss increased from 23 to 35 kg between 18 and 29 degrees C, but its estimated chemical composition remained constant. Pig growth rate was almost constant between 18 and 25 degrees C (241 g/d) and was reduced above 25 degrees C (212 and 189 g/d at 27 and 29 degrees C, respectively). In conclusion, temperatures above 25 degrees C seem to be critical for lactating sows in order to maintain their performance. PMID- 10461992 TI - Phytase improves iron bioavailability for hemoglobin synthesis in young pigs. AB - Dietary phytase supplementation improves bioavailabilities of phytate-bound minerals such as P, Ca, and Zn to pigs, but its effect on Fe utilization is not clear. The efficacy of phytase in releasing phytate-bound Fe and P from soybean meal in vitro and in improving dietary Fe bioavailability for hemoglobin repletion in young, anemic pigs was examined. In Exp. 1, soybean meal was incubated at 37 degrees C for 4 h with either 0, 400, 800, or 1,200 units (U) of phytase/kg, and the released Fe and P concentrations were determined. In Exp. 2, 12 anemic, 21-d-old pigs were fed either a strict vegetarian, high-phytate (1.34%) basal diet alone, or the diet supplemented with 50 mg Fe/kg diet (ferrous sulfate) or phytase at 1,200 U/kg diet (Natuphos, BASF, Mt. Olive, NJ) for 4 wk. In Exp. 3, 20 anemic, 28-d-old pigs were fed either a basal diet with a moderately high phytate concentration (1.18%) and some animal protein or the diet supplemented with 70 mg Fe/kg diet, or with one of two types of phytase (Natuphos or a new phytase developed in our laboratory, 1,200 U/kg diet) for 5 wk. In Exp. 2 and 3, diets supplemented with phytase contained no inorganic P. In Exp. 1, free P concentrations in the supernatant increased in a phytase dose-dependent fashion (P<.05), whereas free Fe concentrations only increased at the dose of 1,200 U/kg (P<.10). In Exp. 2 and 3, dietary phytase increased hemoglobin concentrations and packed cell volumes over the unsupplemented group; these two measures, including growth performance, were not significantly different than those obtained with dietary supplemental Fe. In conclusion, both sources of phytase effectively degraded phytate in corn-soy diets and subsequently released phytate-bound Fe from the diets for hemoglobin repletion in young, anemic pigs. PMID- 10461993 TI - Effect of trimethylamine oxide and betaine in swine diets on growth performance, carcass characteristics, nutrient digestibility, and sensory quality of pork. AB - Two growth experiments and one digestibility experiment were conducted to study the effect of trimethylamine oxide (TMAO) and betaine in swine diets. In Exp. 1, 36 limit-fed pigs averaging 19.1 kg in initial weight were used to study the effect of adding TMAO at 10 g/kg of feed or betaine at an equivalent level of methyl groups (10.5 g/kg feed) to a high-fat (11.3% ether extract) basal diet. Dietary addition of TMAO increased ADG by 61 g/d, reduced number of days to market by 8.3 d (P<.02), and tended (P<.09) to improve gain/feed (G/F) compared with the control diet. Betaine had no effect on growth performance of pigs. Adding TMAO or betaine to diets had no effect on percent carcass fat, percent carcass lean, or dressing percentage. Dietary supplementation of TMAO reduced (P<.05) plasma triacylglycerol level (TAG) compared with the control diet. There was no effect of dietary TMAO or betaine on sensory quality characteristics of pork. In Exp. 2, 48 ad libitum-fed pigs averaging 21.7 kg initial BW and 104.7 kg final BW were used to determine the effect of adding low and intermediate levels of TMAO (1, 2, or 5 g/kg) to diets. Adding 1 g of TMAO increased G/F (P<.01) compared with control pigs. When using orthogonal contrasts, adding 2 g of TMAO reduced (P<.05) P2 backfat thickness and tended to increase (P<.09) lean percentage compared with the control diet. Trimethylamine oxide gave a quadratic effect (P<.05) on plasma TAG levels. Adding 1 and 2 g of TMAO increased plasma TAG, but 5 g of TMAO decreased it compared with the control diet. In Exp. 3, 12 barrows of 42.3 kg average initial BW and 50.0 kg final BW were used to investigate the effect of supplementing diets with 1 g of TMAO and 1.27 g of betaine/kg of feed on apparent total tract nutrient digestibility. The addition of TMAO increased (P<0.03) apparent total tract digestibility of fat (HCl-EE). Betaine had no such effect. Adding TMAO to diets influenced growth performance and carcass quality in a dose-dependent manner. PMID- 10461994 TI - Energy partitioning in growing pigs: the use of a multivariate model as an alternative for the factorial analysis. AB - To quantify the utilization of ME by growing pigs, a factorial analysis method is often used in which the ME intake is regressed on protein (PD) and lipid deposition (LD) rates. The approach has been criticized because there often is a strong correlation between PD and LD, which makes accurate estimation of model parameters difficult. The current study describes a nonlinear multivariate analysis procedure in which PD and LD are the result of variation in ME intake. The approach requires a hypothesis concerning the partitioning of ME intake above maintenance between PD and LD. The method was evaluated using data for growing pigs of different genotypes and sex and weighing between 20 and 107 kg that were offered a diet close to ad libitum. Energy, nitrogen, and fat balances were determined at regular intervals over the growing period. The maintenance energy requirement was expressed as a function of BW (with group-specific parameters) or as a function of muscle and visceral mass. The maintenance energy requirements ranged from 913 to 1,070 kJ ME/((kg BW).60.d) for obese castrates and boars of a synthetic line, respectively. Viscera contributed 1,558 kJ ME/ ((kg tissue).70.d) to the maintenance energy requirement, whereas muscle contributed only 555 kJ ME/ ((kg tissue).70.d). It was assumed that the proportion of ME intake (above maintenance) designated for PD declined linearly with increasing BW. At 20 kg of BW, 49% of ME intake above maintenance was designated for PD in lean genotypes, whereas this was only 34% in obese genotypes. In general, with increasing BW, less energy was designated for PD, but this relationship depended on genotype and sex. Extremely lean male genotypes maintained a constant partitioning of energy between PD and LD for all BW. The energetic efficiencies varied (depending on the model used to express the maintenance requirement) between .58 and .60 for PD and .77 and .82 for LD. Extrapolation of results suggested that animals fed at maintenance energy level would still deposit protein at the expense of body lipid. It is argued that this finding requires nonbiological efficiencies of lipid catabolism and protein synthesis and illustrates the limitation of the maintenance concept for growing animals. The multivariate analysis method proposed here circumvents many of the problems associated with the factorial regression analysis of ME intake on PD and LD. The method can be used to further refine nutritional models describing growth in pigs. PMID- 10461995 TI - Effects of porcine sometotropin on calcium and phosphorus balance and markers of bone metabolism in finishing pigs. AB - Six sets of four littermate barrows initially averaging 75.5 kg BW were equally fed (within blocks) fortified corn-soybean meal diets (1.30% lysine) containing two concentrations of Ca (.50 and 1.00%) and P (.45% and .90%) in a 34-d test. One-half of the pigs were injected with 4 mg of porcine ST (pST)/d. Following a 7 d adjustment period, total collection of feces and urine was performed during two periods (d 1 to 10 and d 20 to 30) for the determination of Ca and P apparent digestibility (absorption) and retention. Pigs were bled after each period (d 10, 20, and 30) for the determination of serum metabolites associated with Ca, P, and bone metabolism. Feed intake for the 30-d period averaged 2,020 g/d. There were no treatment x period interactions, so the absorption and retention data were pooled across periods. The absorption and retention of Ca and P were greater (P<.01) in pigs fed the higher Ca and P levels. Within each Ca and P level, pST reduced (P<.01) fecal Ca and P excretion. Administration of pST did not affect urinary P excretion, but it increased (P<.03) urinary Ca excretion in pigs fed the low-Ca diet. The absorption and retention of Ca and P were increased (P<.01) by pST; however, the increases in Ca retention and P absorption and retention on an absolute basis (g/d) were more pronounced in pST-treated pigs consuming the higher Ca and P diet (interaction, P<.10). Serum concentrations of 1,25 dihydroxyvitamin D3, osteocalcin, and IGF-I on d 10 and 30 were increased (P<.07) with pST administration. However, the increases in 1,25-dihydroxyvitamin D3 and osteocalcin in pST-treated pigs were more pronounced when the lower dietary Ca and P levels were fed (interaction, P<.08). Urinary excretion of hydroxyproline increased (P<.01) with pST administration, but this effect was more pronounced in pST-treated pigs fed the lower Ca and P diet (interaction, P<.09). These results suggest that pST increases the absorption and retention of Ca and P independent of dietary Ca and P level. However, serum measures associated with Ca, P, and bone metabolism in pST-treated pigs were dependent on the Ca and P content of the diet, suggesting an effect of pST on the homeostatic control of Ca, P, and bone metabolism. PMID- 10461996 TI - Effects of dietary levels of selenium-enriched yeast and sodium selenite as selenium sources fed to growing-finishing pigs on performance, tissue selenium, serum glutathione peroxidase activity, carcass characteristics, and loin quality. AB - This research evaluated the efficacy of inorganic and organic Se sources for growing-finishing pigs, as measured by performance and various tissue, serum, carcass, and loin quality traits. A total of 351 crossbred pigs were allotted at an average BW of 20.4 kg to six replicates of a 2x4 factorial experiment in a randomized complete block design. Pigs were fed diets containing Se-enriched yeast (organic) or sodium selenite (inorganic), each at .05, .10, .20, or .30 mg Se/kg diet. A non-Se-fortified basal diet was a ninth treatment group. Five pigs per pen were bled initially and at 30-d intervals with serum analyzed for Se and glutathione peroxidase (GSH-Px) activity. At 55 kg BW, one pig per pen from each of three replicates was killed, and tissues were collected for Se analysis. At 105 kg BW, the remaining pigs in the three replicates were killed, carcass measurements were collected, tissues were analyzed for Se, and loin quality was evaluated for pH, drip loss, and lightness. No performance or carcass measurement benefit resulted from either Se source or dietary Se levels. Pigs had a lower serum Se concentration and GSH-Px activity when the basal diet was fed, but both increased as dietary Se level increased (P<.01). Serum GSH-Px activities were increased by pig age and reached a plateau when the diet contained approximately .10 mg Se/kg (P<.01) at d 30, and 60 of the trial, and at .05 mg Se/kg diet at d 90 of the trial. The organic Se group fed .05 and .10 mg Se/kg had serum GSH-Px activities that tended to be lower than those of pigs fed the inorganic Se source, but GSH-Px activities in both groups were similar at higher Se levels. Tissue Se contents increased linearly as the dietary Se level increased, but the increase was markedly higher when organic Se was fed, resulting in an interaction (P<.01) response. Loin drip loss, pH, and lightness were unaffected (P>.15) by organic Se source or level, but there was a trend for a higher drip loss (P = .11) and a linear (P<.01) increase in loin paleness when the inorganic Se level increased. These results indicate that neither Se source nor Se level had an effect on pig performance or carcass measurements, but organic Se source increased tissue Se concentrations. Inorganic Se may, however, have a detrimental effect on loin quality, as reflected by higher drip loss and a paler color. Using serum GSH-Px activity as the measurement criterion, the supplemental dietary Se requirement did not seem to exceed .10 and .05 mg Se/kg diet for the growing and finishing phases, respectively, when added to a basal diet containing .06 mg Se/kg. PMID- 10461997 TI - Evaluation of selected high-starch flours as ingredients in canine diets. AB - Cereal grains represent 30 to 60% of the DM of many companion animal diets. Once incorporated into a diet, the starch component of these grains can provide an excellent source of ME. However, crystallinity and form of starch are variable and can cause incomplete digestion within the gastrointestinal tract. Diets fed in this experiment included one of six high-starch flours as the main source of carbohydrate. The flours originated from barley, corn, potato, rice, sorghum, and wheat. The diets were extruded and kibbled. Starch fraction concentrations of flours consisted of nearly 100% rapidly digestible starch (RDS) and slowly digestible starch (SDS) combined. Starch fraction concentrations of diets paralleled concentrations in flours. Flours varied widely in concentrations of CP, fat, starch, and total dietary fiber. Ileal OM and CP digestibilities were lowest for the potato flour treatment (74 and 64%, respectively). Ileal and total tract starch digestibilities were different (P<.05) among treatments; however, the starch component of all diets was nearly completely digested (>99%). Total tract digestibility of DM and OM was lowest for sorghum (80 and 84%, respectively) compared to all other diets. Crude protein digestibility was highest for corn (87%). Wet fecal weights tended (P<.08) to be greatest for dogs fed the barley treatment (175 g/d). However, dry fecal weights (dried at 55 degrees C) were greatest for dogs consuming the sorghum diet (51 g/d). Fecal scores were consistently greater (i.e., looser stools) for the barley treatment. Any of these flours could be used without negative effects on digestion at either the ileum or in the total tract. Fecal consistency data for dogs consuming the barley treatment indicate that diets containing large amounts (>50%) of barley may not be advantageous for dog owners who house their animals indoors for most of the day. PMID- 10461998 TI - Abortifacient effects of a unique class of vasoactive lipids from Pinus ponderosa needles. AB - Pinus ponderosa needle (PN) ingestion by late pregnant cows results in decreased uterine blood flow, premature parturition, and retained placentae. Further, plasma from PN-fed cows increases caruncular arterial tone (i.e., induces prolonged contraction) in an isolated perfused bovine placentome. A novel class of vasoactive lipids was isolated and identified using a bovine placentome assay guided fractionation of CH2Cl2 extracts of PN. Placentome perfusion tests indicated that 1-12-dodecanedioyl-dimyristate (14-12-14) was the most potent of the PN lipids for increasing caruncular arterial tone. Late pregnant guinea pigs (GP) were used to evaluate the abortifacient activity of these vasoactive lipids. In Study 1, on d 50 of gestation, part of the control diet was replaced with chopped PN (Diet A) or chopped PN subjected to sequential extraction with diethyl ether (Et2O; Diet B); Et2O and CH2Cl2 (Diet C); and Et2O, CH2Cl2, and methanol (Diet D). The GP on Diets A and B exhibited shorter (P<.01) gestation lengths and reduced (P<.01) pig birth weights than GP on the control diet or Diets C and D. Further, only GP on Diets A and B exhibited retained placentae. In Study 2, on d 50 of gestation, part of the control diet was replaced with chopped PN that had been subjected to exhaustive CH2Cl2 extraction and then infiltrated with either CH2Cl2 alone (Diet E), CH2Cl2 containing 14-12-14 (Diet F), or CH2Cl2 containing isocupressic acid (Diet G); then solvents were evaporated. The GP consuming Diet F had shorter (P<.05) gestation lengths and reduced (P<.05) pig birth weights than did GP consuming Diets E or G. The GP consuming Diet F also exhibited a high incidence of retained placentae. These data provide evidence that a unique class of vasoactive lipids in PN exhibit abortifacient activity in guinea pigs. PMID- 10461999 TI - Field method for monitoring blood glucose in beef cattle. AB - The purpose of this study was to determine the applicability of the Accu-Chek Easy (ACE) human self-monitoring system for monitoring glycemic status in cattle. The ACE method was compared with the Yellow Springs Instrument (YSI) analytical laboratory method in two studies. A preliminary study (62 samples) and a primary study (434 samples) involved a nine-fold range and a 10-fold range, respectively, of glucose concentrations obtained during the acute phase response of growing beef cattle to injections of varying dosages of endotoxin. The ACE monitoring method compared with the YSI analytical method resulted in similar patterns of glucose concentration change, similar ranking of glucose means across endotoxin dosages during hyper-and hypoglycemia, and a close relationship between paired YSI and ACE concentrations from common samples. The ACE method identified all nine animals that displayed hypoglycemic distress during the acute phase response to endotoxin injection. The relationship between the YSI analytical method and the ACE monitoring method was found to be nonlinear (YSI = -38.2+13.6.ACE.50; R2 = .99; Sy.x = 7.3 mg/dL), and the use of this equation to predict YSI values from ACE values in an independent data set resulted in linearity when YSI was regressed on the predicted YSI values (YSI = -.78+1.00. Predicted YSI; R2 = .87; Sy.x = 6.9 mg/dL). Even though variation seemed greater for ACE than for YSI, we concluded that a system developed for human self-monitoring of blood glucose, such as the ACE, can be used to monitor the glycemic status of cattle. PMID- 10462000 TI - The effect of spermatozoa and seminal plasma on leukocyte migration into the uterus of gilts. AB - Yorkshire x Landrace gilts were used to determine the effect of spermatozoa and seminal plasma on postbreeding uterine leukocyte influx. Estrus detection was performed with a boar at 12-h intervals following synchronization with 400 IU eCG and 200 IU of hCG. All gilts were AI once, 24 h after the detection of estrus following random assignment to a 2x2x3 factorial arrangement of treatments (sperm or sperm-free AI doses), AI dose medium (seminal plasma or PBS), and lavage time following AI. Gilts were treated with sperm (5x10(9) spermatozoa; SPZ; n = 30) or sperm-free (SF; n = 30) doses containing either 100 mL of seminal plasma (SP; n = 15/treatment) or PBS (n = 15/treatment). Uterine lavage was performed once on each gilt (n = 20/time) at one of three times after AI (6, 12, or 36 h) to determine the total number of uterine leukocytes. The leukocytes consisted predominately (92 to 99%) of polymorphonuclear neutrophilic granulocytes (PMN). There was an AI x medium interaction on uterine PMN numbers. The number of uterine PMN recovered from gilts inseminated with sperm suspended in PBS was greater than the number of PMN recovered from the uterine lumen of gilts inseminated with sperm in SP, SP alone, or PBS alone (P<.05). Furthermore, SP accelerated the rate of uterine clearance when suspended with sperm cells during the first 36 h following AI (P<.05). These results indicate that seminal plasma suppresses PMN migration into the uterus following breeding and enhances the rate of disappearance of uterine inflammation. PMID- 10462001 TI - Twice daily suckling but not milking with calf presence prolongs postpartum anovulation. AB - Two experiments were conducted to determine whether milking beef cows two or five times daily in the presence or absence of their own nonsuckling calves would alter postpartum interval to first ovulation. Multiparous Angus x Hereford cow calf pairs were assigned randomly between 13 and 18 d postpartum to treatments for 4 wk. In Exp. 1, pairs were assigned to six treatments: 1) calf was weaned permanently from its dam (CW; n = 9); 2) same as CW, but dam was milked twice daily (CW+2xM; n = 9); 3) calf was present continuously with its dam but restricted from contact with the udder (CR; n = 9); 4) same as CR, but dam was milked twice daily (CR+2xM; n = 9); 5) same as CR, but calf was allowed to suckle twice daily (CR+2xS; n = 8); and 6) calf was present continuously with its dam and suckled ad libitum (CP; n = 9). The interval from onset of treatments to first postpartum ovulation was shorter (P<.05) in the CW (14.1+/-3.1 d), CR (14.2+/-3.1 d), CW+2xM (13.0+/-3.1 d), and CR+2xM (17.2+/-3.1 d) than in the CP (34.7+/-3.1 d) and CR+2xS (33.9+/-3.3 d) treatments. Daily milk yield during treatment was greater (P<.01) for CR+2xM cows (7.1+/-.6 kg) than for CW+2xM cows (3.5+/-.6 kg). In Exp. 2, cow-calf pairs were assigned to three treatments: 1) CR+2xM (n = 10); 2) same as CR+2xM but cows were milked five times daily (CR+5xM; n = 10); or 3) CP (n = 10). The interval to first postpartum ovulation was shorter (P<.05) in the CR+2xM (23.6+/-3.5 d) and CR+5xM (26.1+/-3.7 d) treatments than in the CP (37.7+/-3.7 d) treatment. Daily milk yield during treatment was greater (P<.05) for CR+5xM cows (7.7+/-.6 kg) than for CR+2xM cows (6.4+/-.6 kg) by 17%. We conclude that suckling twice daily was sufficient to prolong postpartum anestrus as much as suckling ad libitum. Furthermore, milk removal by suckling, but not by milking two or five times daily, even in the presence of the cow's own nonsuckling calf, is essential to prolong postpartum anovulation. PMID- 10462002 TI - Effect of duration of dominance of the ovulatory follicle on onset of estrus and fertility in heifers. AB - In cattle, prolonged progestogen treatments following luteolysis result in persistent dominant follicles (DF) that are associated with precise onset of estrus but marked reductions in pregnancy rate (PR). The aim was to determine whether increasing duration of dominance of the ovulatory follicle in heifers affected 1) precision of onset of estrus and 2) the timing and nature of the decline in PR. In Exp. 1, duration of dominance of the ovulatory follicle was controlled by causing corpus luteum (CL) regression at emergence of the second follicle wave (mean duration of dominance of 2.1+/-.3 d, Dm2, n = 11) or first day of dominance of the second DF of the cycle; the latter was combined with insertion of a 3-mg norgestomet ear implant for 2 to 10 d to maintain the second DF for 4 (Dm4, n = 32), 6 (Dm6, n = 19), 8 (Dm8, n = 49), 10 (Dm10, n = 28), or 12 d (Dm12, n = 20). Heifers detected in estrus were inseminated approximately 12 h later with frozen-thawed semen. Durations of dominance of the ovulatory follicle of up to 8 d did not affect (P>.05) PR (Dm2 8/9, Dm4 19/28, Dm6 14/18, and Dm8 34/48 heifers pregnant), but PR in Dm10 heifers (12/23 heifers pregnant) was reduced (P = .05) compared with Dm2 heifers; PR in Dm12 heifers (2/17 pregnant) was less compared with all other treatments (P<.01). Fitting a logistic regression model to the pooled PR data to examine the trend in PR showed that extending the duration of dominance from 2 to 9 d and from 10 to 12 d resulted in a predicted decline in PR of 10 to 25% and a further decline of 35 to 75%, respectively. Onset of estrus was delayed in heifers assigned to Dm4 treatment relative to all other treatments (P<.001); it was less variable than that for heifers on Dm6, Dm8, and Dm10 treatments (P<.1). In Exp. 2, heifers received a PGF2alpha analogue and a norgestomet implant on d 12 of the cycle for 3 or 7 d to give approximate durations of dominance of the preovulatory follicle of 2 to 4 d (Dm2-4, n = 29) or 6 to 8 d (Dm6-8, n = 24), respectively. The PR did not differ (P>.05) between heifers on Dm2-4 (22/29) and Dm6-8 (15/24) treatments, but the interval to onset of estrus was delayed (P<.05) by 7 h in the Dm2-4 heifers. In conclusion, restricting the duration of dominance of the preovulatory follicle to < or =4 d at estrus, results in a precise onset of estrus and a high PR following a single AI at a detected estrus. PMID- 10462003 TI - Effects of prepartum supplementary fat and muscle hypertrophy genotype on cold tolerance in newborn calves. AB - Effects of feeding pregnant dams supplemental dietary fat during the last 55 d of gestation on cold tolerance of newborn crossbred calves with (Piedmontese cross, P, n = 15) or without (Hereford cross, H, n = 16) the muscle hypertrophy allele was determined. Primiparous F1 dams gestating F2 calves of the respective breeds were assigned randomly within breed to receive gestation diets containing either 2.2 (Low Fat; LF) or 5.1% fat (High Fat; HF). Safflower (Carthamus tinctorius L.) seeds containing 37% oil with 79% linoleic acid were the supplemental fat source in diets formulated to be isocaloric-isonitrogenous. At parturition, calves were separated from their dams, fed 38 degrees C pooled dairy cow colostrum (30 mL/kg BW), muzzled to prevent suckling, and returned to their dams in a heated (22 degrees C) room for 3.5 h. At 4 h of age (birth = 0 h), a catheter was inserted into the jugular vein. At 5 h of age, calves were placed in a 0 degrees C room for 140 min, and rectal temperatures and blood samples were obtained at 10- and 20-min intervals. Blood was assayed for cortisol and glucose. Rectal temperature was affected by diet (P<.05), time, diet x time, and breed x time (P<.01 for time and the interactions). Cortisol and glucose concentrations were not affected by diet, breed, or the diet x breed interaction, but they were affected by time, breed x time (both P<.01), and diet x time (P = .06). Calves from HF dams had higher rectal temperatures than calves from LF dams, and the HF calves maintained higher rectal temperatures throughout cold exposure. Cortisol concentrations were lower (P = .06) in calves from HF dams, and these calves had more (P = .06) glucose available for metabolic heat production than calves from LF dams. Piedmontese-cross calves maintained higher (P<.01) rectal temperatures and had higher cortisol and glucose (both P<.01) concentrations than did H-cross calves. We conclude that feeding dams supplemental fat during late gestation increased heat production in newborn calves and potentially could increase calf survival; calves with muscle hypertrophy may have a different ratio of shivering vs nonshivering thermogenesis due to differences in body composition or relationships among uncoupling proteins. PMID- 10462004 TI - Production and lambing rate of blastocysts derived from in vitro matured oocytes after gonadotropin treatment of prepubertal ewes. AB - The aim of this study was to evaluate the effect of gonadotropin treatment on the in vitro maturation, blastocyst production, and developmental potential to term of oocytes collected from Sardinian neonatal and prepubertal ewes at 4 to 6 wk of age. Cumulus-oocyte complexes were recovered at 24 h after withdrawal of a 1/6th size progestagenated pessary from the donors, of which each received 120 IU FSH/LH and 400 IU PMSG in a single dose 36 h before sponge removal. Treated donors produced a greater (P<.01) number of oocytes per animal (86.2 +/-7.9) compared with slaughterhouse (untreated) prepubertal ewes (55.5+/-6.1) of the same age or with treated neonatal ewes (6.1+/-0.7) 10 d old. During oocyte maturation, there were no differences in the percentage of germinal vesicle break down (78.08 vs. 74.24), metaphase I (89.13 vs. 87.18), and metaphase II (77.91 vs. 76.38) when evaluated after 8, 14, and 24 h of maturation, respectively, between oocytes from treated and slaughterhouse (untreated) prepubertal ewes. The embryo cleavage (71.1 vs. 73.7) and blastocyst rates (22.2 vs. 19.8) were similar in the treated and the untreated prepubertal ewes after transfer of in vitro matured oocytes into ligated oviducts of temporary recipients. The in vitro viability rates of vitrified blastocysts (81.2 vs. 76.9) and the in vivo survival rates (46.1 vs. 50.0) of embryos derived from in vitro matured and in vivo fertilized oocytes showed no difference. The data suggest that gonadotropin treatment increases oocyte production per animal but has no effect on oocyte quality because embryo production and lambing rates of blastocysts derived from in vitro matured oocytes were not markedly different from those derived from untreated prepubertal ewes of the same age. PMID- 10462005 TI - Effects of the thymic peptide thymulin on in vitro and in vivo testicular steroid concentrations in white composite and Meishan boars. AB - Immuno-peptides may have positive or negative effects on gonadal steroidogenesis, but few have been tested outside of rodent species or in vivo. In Exp. 1, thymulin, a secreted nonapeptide of the thymus, was incubated (1, 10, 100, or 1,000 ng/mL) with testicular minces (sampled at 3, 6, or 12 h) from Chinese Meishan boars of high gonadotropin/testicular steroidogenic function (n = 8) and White composite boars of European origin (n = 8 ) to test the hypothesis that thymulin could augment hCG stimulation of testicular androgen concentrations. Thymulin alone had few effects on androgen concentrations (testosterone, dehydroepiandrosterone+dehydroepiandrosterone sulfate [DHEA+DHEASO4]) in Meishan boar testicular incubates. In minces from White composites incubated with thymulin, testosterone concentrations were generally below control values (P<.05), but DHEA+DHEASO4 concentrations were unaffected. Thymulin had no effect on estrone concentrations in testicular incubates of White composite boars but stimulated estrone concentrations in Meishan testicular incubates. Thymulin plus hCG increased testosterone (3 and 6 h of incubation; P<.05) but not DHEA+DHEASO4 concentrations in White composite testicular incubates. Thymulin plus hCG did not alter androgen or estrogen concentrations from control values in Meishan testicular incubates. In Exp.2 with a protocol similar to that of Exp. 1 for testicular minces from White composite boars (n = 30), thymulin increased testosterone concentrations during the early incubation period (1 to 3 h; P<.05) and depressed testosterone concentrations at later times (6 h; P<.05). Thymulin synergized with hCG in stimulating increases in testosterone and DHEA+DHEASO4 concentrations (P<.05) but had no effect on estrone concentrations in vitro. Thymulin was tested in vivo in boars from three genetic lines selected for high, medium, or normal circulating LH concentrations (Meishan, select White composites, and control White composites, respectively). Injection of thymulin i.v. (4.4, 44.4, or 444.4 ng/kg BW) generally increased circulating testosterone concentrations (2 to 3 h later; P<.01), but the response was dependent on the boar's general circulating LH concentrations and dose of thymulin. Overall results from these studies support the hypothesis of a thymulin augmentation of LH stimulation of androgen increases in vitro and in vivo in the testis of boars. PMID- 10462006 TI - Diet selection by steers using microhistological and stable carbon isotope ratio analyses. AB - Two methods of determining diet botanical composition, microhistological (MH), and stable carbon isotope ratio (CR) analyses were used to determine botanical composition of ingesta and fecal grab samples in steers grazing rhizoma peanut mixed tropical grass pastures. Three pastures were used over two grazing seasons, 1992 and 1993, in Brooksville, FL. A weighted-disc double-sampling technique was used to determine forage mass and botanical composition, percentage of rhizoma peanut (Arachis glabrata), grass (Paspalum notatum and Cynodon dactlyon), and forb (primarily Chenopodium ambrosioides) on offer every 28 d throughout the grazing seasons. There was an effect of sampling date (P<.001), sampling date x pasture (P<.001), and sampling date x year (P<.001) on forage mass on offer. There was a pasture x year x sampling date interaction (P<.001) for all botanical components. In 1992 and 1993, using cannulated steers sampled every 56 d, there were interactions with year for rhizoma peanut and forb (P<.05), but not for grass with MH analysis (components: rhizoma peanut, grass, and forb). Ingesta and fecal rhizoma peanut (r = .73 and .92 for 1992 and 1993, respectively) and ingesta and fecal forb (r = .86 and .98 for 1992 and 1993, respectively) were positively correlated (P<.001). Ingesta and fecal grass were positively correlated (r = .52, P<.001), but the correlation was not as high. With the CR analysis (components: Calvin cycle [C3] plants and C4-dicarboxylic acid pathway [C4] plants), ingesta and corrected fecal (corrected for in vitro organic matter digestibility [IVOMD]) C3 plants were positively correlated (r = .62; P<.001). Diet composition of fecal grab samples from noncannulated steers, collected on the same sampling schedule as for hand-clipped pasture samples, differed at times due to the complexity of the sward (both rhizoma peanut and forb constituted a single component, C3, in the CR analysis). Based on these results, if there is a substantial contribution of forb to the diet, fecal microhistological analysis may be more informative than fecal carbon ratio analysis for estimating diet selection by cattle grazing tropical pastures. PMID- 10462007 TI - Effect of virginiamycin on ruminal fermentation in cattle during adaptation to a high concentrate diet and during an induced acidosis. AB - The objective of Exp. 1 was to compare the effects of virginiamycin (VM; 0, 175, or 250 mg x animal(-1) x d(-1)) and monensin/tylosin (MT; 250/ 90 mg x animal(-1) x d(-1)) on ruminal fermentation products and microbial populations in cattle during adaptation to an all-concentrate diet. Four ruminally cannulated, Holstein steers were used in a 4x4 Williams square design with 21-d periods. Steers were stepped up to an all-concentrate diet fed at 2.5% of BW once daily. Ruminal pH, protozoal counts, and NH3-N and VFA concentrations generally were unaffected by VM or MT. Mean counts of Lactobacillus and Streptococcus bovis were lower (P<.05) for VM-treated compared with control or MT-treated steers. Both VM and MT prevented the increase in Fusobacterium necrophorum counts associated with increasing intake of the high-concentrate diet observed in the control. The objective of Exp. 2 was to compare the effects of VM and MT on ruminal pH, L(+) lactate and VFA concentrations, and F. necrophorum numbers during carbohydrate overload. Six ruminally cannulated Holstein steers were assigned randomly to either the control, VM (175 mg/d), or MT (250 + 90 mg/d) treatments. Acidosis was induced with intraruminal administration of a slurry of ground corn and corn starch. The VM and MT premixes were added directly to the slurry before administration. Carbohydrate challenge induced acute ruminal acidosis (pH was 4.36 and L (+) lactate was 19.4 mM) in controls by 36 h. Compared with the controls, steers receiving VM or MT had higher (P<.05) ruminal pH, and the VM group had a lower (P<.05) L (+) lactate concentration. Fusobacterium necrophorum numbers initially increased in VM- and MT-administered steers. In the control steers, F. necrophorum was undetectable by 36 h. Virginiamycin seemed to control the growth of ruminal lactic acid-producing bacteria and, therefore, has the potential to moderate ruminal fermentation in situations that could lead to rapid production of lactic acid. PMID- 10462008 TI - Effect of wheat and corn variety on fiber digestion in beef steers fed high-grain diets. AB - Six Salers steers, fitted with ruminal and duodenal cannulas, were used in a double 3x3 Latin square design to assess the depressive effect of the nature of wheat, flint corn, and dent corn on fiber digestion in animals fed high concentrate diets, and to determine the mechanisms involved in these negative digestive effects. Diets were balanced to be equal in starch content (47.7+/ 2.3%). The three cereals were characterized by ruminal starch digestibilities of 86.6, 60.8, and 34.8% for the wheat, dent corn, and flint corn, respectively. Ruminal digestion of NDF was lower with wheat- than with corn-based diets (49.4 vs. 55.2%; P<.001), and with dent corn than with flint corn (53 vs. 57.3%; P<.01). Degradability of hay in nylon bags was not affected by the grain source in the diet (P>.1). The mean retention time of forage particles in the rumen was similar between wheat and corn diets (P>.1), but it was lower for steers fed dent corn than for those fed flint corn (P<.05). Most fibrolytic activities of the solid-associated microorganisms were lower (P<.05) in animals fed wheat than in those fed corn. Differences in fibrolytic activities of the solid-associated microorganisms between the two corn genotypes were not statistically significant (P>.1), but activities of all fibrolytic enzymes were lower (P<.05) with the dent than with the flint corn diet. Protozoal number in ruminal fluid was lower in animals receiving wheat than in those fed corn (177 vs. 789x10(3)/mL; P<.001) and was related to the high ruminal acidity (P<.01) of the wheat diet. Large modifications in the rumen microbial ecosystem between the two corn genotypes were not visible in protozoal numbers or pH. Total-tract digestion of NDF was the same for wheat and for corn diets, averaging 55% for the three diets. A postruminal compensation of NDF digestion (14% of the total tract NDF digestion) seemed to occur with the wheat diet. The lack of any postruminal NDF digestion (0%) with the two corn diets may suggest negative digestive interactions in the hindgut similar to those in the rumen. PMID- 10462009 TI - Effects of underfeeding and refeeding on weight and cellularity of splanchnic organs in ewes. AB - We assessed the effects of a long and severe period of underfeeding, followed by a rapid refeeding with a high-concentrate diet, on weight, protein mass, and cellularity of the splanchnic organs in adult ewes. Twenty-four ewes, allocated to four groups of six, were fed a forage diet (50% regrowth of natural grassland hay and 50% wheat straw) either at maintenance (groups M and MO) or at 40% maintenance (groups U and UO) for 78 d. Groups M and U were then slaughtered, and groups MO and UO were subsequently overfed a high-concentrate diet (52% hay, 20% barley, 16% rapeseed meal, 4% fish meal, and 8% Megalac) at 236% maintenance for 26 d before being slaughtered. During the experiment, feed was adjusted to maintain feed supply at a constant percentage of animal requirements. After slaughter, fresh weight, dry weight, and protein mass of the reticulorumen, omasum, abomasum, small intestine, large intestine, and liver were measured. Cellularity was assessed from nucleic acids and protein contents for both ruminal mucosa and muscular-serosa layers, jejunum, and liver. The concentrations of ubiquitin and cathepsin D mRNA were measured in ruminal mucosa and muscular serosa layers and in jejunum. Underfeeding decreased protein mass of splanchnic organs, especially in liver (-29%) and reticulorumen (-39%). Refeeding previously underfed animals increased protein mass of liver (+102%) and small intestine (+59%). No carry-over effect of the previous level of intake (UO vs. MO) was observed on the protein mass of splanchnic tissues after 26 d of refeeding. Variations in liver mass were mainly due to hypertrophy, as determined by the protein:DNA ratio, whereas variations in small intestinal mass were mainly due to hyperplasia, as determined by the amount of DNA. By contrast, changes in rumen mass associated with increasing ME intake seemed to be related to hypertrophy in the muscular-serosal component and hyperplasia in the epithelial component. The concentrations of ubiquitin and cathepsin D mRNA in the rumen and jejunum were not modified by feeding level, demonstrating that the expression of these genes for proteolytic enzymes was unchanged under these conditions. PMID- 10462010 TI - Comparison of models estimating digesta kinetics and fecal output in cattle from fecal concentrations of single-dosed markers of particles and solutes. AB - Fecal concentrations of chromium and cobalt, following a single labeling dose of Cr-mordanted fiber and Co-EDTA to the rumen, were obtained from two experiments: one with beef cows fed two diets before and after parturition, and one with growing bull calves fed two diets of high and low fill volume. These data were used to compare three optional models that estimated the whole digestive tract kinetics of particles and solutes and fecal output, with a fourth model that estimated particle kinetics only. The first model (M1) assumes separate routes for particles and solutes, with two bypass fluxes (i.e., simultaneous fluxes from one pool to more than one other pool) in the particle route and one in the solute route. The second model (M2) is similar to M1, but allowance is made for some of the particles to pass to the solute route. The third model (M3) assumes that most of the kinetic variables in the solute route are identical to the correspondent variables in the particle route. The fourth model (M4) assumes an unspecified number of sequential compartments with constant increase of the outflow rate from each compartment to the next one, without allowance for any bypass fluxes. All the models could fit all the data sets. Goodness of fit was the best with M2 and the worst with M3. Goodness of fit of the particle curve with M2 was comparable to that of M4. Model M1 estimated the shortest and M3 estimated the longest total retention time (TRT) for particles (72.8, 85.0, and 91.2 h for M1, M2, and M3, respectively), and the partition of retention time between the different pools differed among models. There were no significant differences among models in their estimates of solute TRT (30.2, 31.5, and 30.8 h for M1, M2, and M3, respectively). Fecal output estimations were similar among models, all of them overestimating the predetermined measurements by 9.5 to 13%. The r2 of the linear regression of the estimated on the determined fecal output was .74, .75, and .70 for M1, M2, and M3, respectively. PMID- 10462011 TI - Effects of laidlomycin propionate and monensin on the in vitro mixed ruminal microorganism fermentation. AB - The objective of this study was to compare the effects of laidlomycin propionate and monensin on the in vitro fermentation of ground corn, Trypticase, or alfalfa hay by mixed ruminal microorganisms. Ruminal fluid was collected from two steers fed 9.27 kg DM of a high-concentrate (62.2% ground corn and 17.4% cottonseed hulls) diet per day and composited. In the first study, no ionophore was included in the diet; the diet in the second study contained 11.1 g of laidlomycin propionate per ton of feed. The animals were allowed an adjustment period of 14 d for each dietary treatment before samples were collected. When ruminal fluid from unadapted animals was used, both monensin and laidlomycin propionate decreased (P<.05) CH4 concentration and the acetate:propionate ratio with ground corn and alfalfa hay. Monensin reduced (P<.05) in vitro dry matter disappearance of alfalfa and increased (P<.05) final pH in the ground corn and alfalfa hay fermentations. Both laidlomycin propionate and monensin decreased (P<.05) concentrations of acetate, propionate, isobutyrate, isovalerate, CH4, and NH3 in Trypticase fermentations. When ruminal fluid from adapted animals was used, both ionophores still reduced the concentrations of most fermentation products. However, there was generally less inhibition compared with fermentations inoculated with unadapted mixed ruminal microorganisms. In the presence of 5 mM maltose, mixed ruminal bacteria produced high concentrations (10 to 11 mM) of lactate, and addition of both ionophores to these fermentations was effective in reducing (P<.05) lactate production. In conclusion, laidlomycin propionate alters the mixed ruminal microorganism fermentation in a manner similar to monensin, but, at the concentrations used in this study, monensin seemed to be a more potent inhibitor. PMID- 10462012 TI - Rapid communication: human melanocortin receptor sequences reveal a SacI restriction fragment length polymorphism at the porcine adrenocorticotropic hormone receptor (MC2R) locus. PMID- 10462013 TI - Rapid communication: three unique restriction fragment length polymorphisms of EcoRI, PvuII, and ScaI digested mitochondrial DNA of Bactrian camels (Camelus bactrianus ferus) in China. PMID- 10462015 TI - Prognostic significance of the pattern of multivessel spasm in patients with variant angina. AB - Multivessel spasm in variant angina is believed to be a major prognostic factor. Three patterns of multivessel spasm have been detected: (1) spasm at different sites on different occasions (migratory spasm); (2) spasm sequentially affecting 2 different sites (sequential spasm); and (3) simultaneous spasm at more than 1 site (simultaneous spasm). The present study investigated the prognosis based on this factor for variant angina without fixed coronary stenosis and examined the influence of multivessel spasm on cardiac events. Twenty-six patients were diagnosed as having variant angina without fixed coronary stenosis using 12-lead 24-h ECG recording system and coronary cineangiography. These patients were followed up prospectively for 57.1+/-7.6 months. Of the 26 patients 13 had single vessel spasm, 6 had migratory multivessel spasm angina, and 7 showed sequential and/or simultaneous multivessel spasm angina. The survival free of serious cardiac events and of all cardiac events was significantly lower for patients with sequential and/or simultaneous multivessel spasm than for those with migratory multivessel spasm (p<0.05, p<0.05), whereas for patients with migratory multivessel spasm the difference comparison with single-vessel spasm did not attain statistical significance (p = ns, p = ns). The results of this study suggest that there seems to be a high-risk subgroup (i.e., sequential and/or simultaneous multivessel spasm) among patients with variant angina. PMID- 10462014 TI - Changes in endothelium-derived vascular regulatory factors during dobutamine stress-induced silent myocardial ischemia in patients with Kawasaki disease. AB - The changes in endothelium-derived vascular regulatory factors during dobutamine (DOB)-induced myocardial ischemia (MI) were investigated in 21 patients with Kawasaki disease aged from 11 months to 18 years. They were classified into an ischemia group (8 patients) and a non-ischemia group (13 patients) based on the results of 99mTc myocardial scintigraphy and DOB stress 99mTc myocardial scintigraphy. In the ischemia group, MI was relatively mild, because there were ischemic changes on the electrocardiogram and no significant symptoms during DOB stress. Catheters were positioned near the orifice of the coronary artery (Ao) and at the coronary sinus (CS). Hemodynamics and the blood concentrations of lactic acid and endothelin-1, as well as NO3-, 6-keto-prostaglandin F1alpha, and thromboxane B2, (which are inactive metabolites of nitric oxide, prostaglandin I2 and thromboxane A2, respectively), were measured at rest and after DOB stress (maximum dose: 30 microg x kg(-1) x min(-1)). The CS/Ao ratio was determined for all parameters. The rate-pressure product, an index of work load, and the cardiac index were significantly increased by DOB stress in both groups. Coronary angiography showed no vasospasm of the epicardial coronary arteries before or after DOB stress in either group. The plasma concentrations of endothelin-1 and 6 keto-prostaglandin F1alpha were significantly increased after DOB stress in the ischemia group, but the serum concentration of NO did not increase. The lack of an increase in NO production during DOB stress may have contributed to the worsening of MI in patients with Kawasaki disease. PMID- 10462016 TI - Acute performance of steroid-eluting screw-in leads for atrial free wall pacing. AB - The aim of this study was to clarify the acute performance of steroid-eluting screw-in leads in comparison with that of nonsteroid screw-in leads for atrial free wall pacing. In 114 cases (68 males, 46 females, average age 70 years) with atrial free wall pacing by screw-in leads, pacing thresholds and P-wave amplitudes were compared at the time of implantation and 1 week later between 68 cases of nonsteroid and 46 cases of steroid-eluting screw-in leads. No significant differences were seen between the 2 groups at implantation in either voltage or current thresholds measured at pulse widths of 0.1, 0.3, 0.6, 1.0, 2.0 ms, or P-wave amplitudes. Pulse width thresholds at outputs of 2.5 V and 5.0 V were significantly lower for steroid leads 1 week after implantation (2.5 V: 0.34+/-0.27 ms nonsteroid vs. 0.12+/-0.08 ms steroid, p<0.001; 5.0 V: 0.12+/-0.08 ms nonsteroid vs. 0.06+/-0.02 ms steroid, p<0.01). P-wave amplitudes after 1 week were significantly higher for steroid leads (2.6+/-0.7 mV nonsteroid vs 3.0+/-1.2 mV steroid, p<0.001). Threshold rise, including pacing failure, was observed in 15 (22%) of the non-steroid leads, but in only 1 (2%) of the steroid leads. In conclusion, steroid-eluting screw-in leads suppress the acute rise of pacing thresholds in the right atrial free wall and their acute performance is better than that of non-steroid leads. These results suggest that appropriate low-output atrial pacing is feasible immediately after implantation. PMID- 10462017 TI - Significance of exercise QT dispersion in patients with coronary artery disease who do not have exercise-induced ischemic ST-segment changes. AB - The poor sensitivity and the poor predictive value of ST-segment depression have limited the usefulness of the exercise electrocardiogram (ECG) in the diagnosis and evaluation of coronary artery disease (CAD). The QT dispersion (QTD), recorded as the difference between maximal and minimal QT intervals on a 12-lead exercise ECG, is sensitive to myocardial ischemia and may improve the accuracy of exercise testing in patients with CAD who do not show an ST-segment depression. Exercise ECGs were analyzed in 50 subjects who had undergone coronary angiography for clinical indications. None of them showed an ST-segment depression during or after exercise: There were 25 patients with significant coronary artery stenosis and 25 without significant stenosis. The QTD measured before, immediately after, and 1 min after exercise was similar in the 2 groups. The QTD at 3 and 5 min after exercise was significantly greater in patients with CAD than in the controls, and the most marked difference in QTD was observed at 3 min after exercise. A QTD at 3 min after exercise of >60 ms had a sensitivity of 80% and specificity of 88% regarding the diagnosis of CAD. When a deltaQTD (post-exercise QTD minus QTD at rest) at 3 min after exercise of >0 ms was added to a QTD of >60 ms as a condition for positivity, the specificity increased to 96%. QTD measured at 3 min after exercise increases the accuracy of exercise testing in patients with CAD who do not show an ST-segment depression. PMID- 10462018 TI - Morphologic correlation between atherosclerotic lesions of the carotid and coronary arteries in patients with angina pectoris. AB - The morphology of atherosclerosis between the carotid and coronary artery systems was studied in 63 patients with ischemic heart disease to determine if there was a correlation with coronary heart disease. The sclerotic lesions of the carotid and coronary artery systems were imaged with ultrasonography and coronary arteriography, respectively, and divided into 4 types. Hemodynamic variables, serum lipid levels, and serum uric acid concentration were not different among the groups, but the serum C-reactive protein (CRP) concentration in patients without significant atheroma in the carotid artery system was lower than the mean concentration of the other 3 groups with carotid atheroma. The morphological stability of carotid arterial plaques correlated well to coronary artery stenosis. Morphologically unstable plaques of the carotid artery predicted unstable forms of coronary obstruction with a sensitivity of 68%, specificity of 85%, predictive power of 72% and a likelihood ratio of 4.5. These results suggest that ultrasonic examination of the carotid artery is useful for predicting the presence or absence of unstable lesions in coronary arteries. PMID- 10462019 TI - Comparisons between hemodynamics, during and after bathing, and prognosis in patients with myocardial infarction. AB - The purpose of this study was to establish the safest way to bathe patients with myocardial infarction (MI) through measuring the hemodynamics during and after bathing. Seventy patients with MI were bathed supine in a Hubbard tank filled with 42 degrees C tap water for 5 min. The subjects were divided into 2 groups depending on their hemodynamic values 10 min after bathing: pulmonary capillary wedge pressure unchanged even after bathing (group A), and decreased pressure after bathing (group B). The left ventricular ejection fraction of group B was significantly higher than that of group A: 53.6% vs. 39.7%, respectively (p<0.01). The physical work capacity of group B was significantly higher at 5.6 METs, than that of group A with 4.5 METs (p<0.05). During the average of their 37 month follow-up period, there were 3 cardiac events in group B and 6 in group A. There were 2 cardiac events during bathing, both of which occurred in group A. When patients with MI take a bath, it is essential to closely monitor them, especially to those patients with lower cardiac function, because they have a higher possibility of a cardiac event. PMID- 10462020 TI - Should use of the internal thoracic artery be avoided under conditions of low free flow? Postoperative hemodynamic assessment using pulsed Doppler echocardiography. AB - There are cases in which it is thought advisable to avoid the use of the left internal thoracic artery (LITA) in coronary artery bypass grafting (CABG) due to its low free flow (FF). However, even though the LITA flow is very low, anastomosis without any further maneuvers intraluminally is recommended. The present study investigated the clinical results of CABG, using a LITA with low FF. The 60 cases of CABG were divided into 2 groups: (i) Group L (n = 23), in which LITA FF was less than 20 ml/min; and (ii) Group H (n = 37) in which it was more than 20 ml/min. A comparative study on the basis of coronary angiography and pulsed Doppler echocardiography was performed. In both groups, no LITA graft occlusion was identified, and the 'string sign' was also absent. In the LITA blood waveform, all cases exhibited a biphasic pattern with a higher mid diastolic and a lower end-systolic component. There were no significant differences in the LITA flow diastolic peak velocity, velocity time integrals and the diastolic/systolic peak velocity ratios. These results suggest that the LITA can be used for CABG even when the free flow is less than 20 ml/min. PMID- 10462021 TI - In vitro examination of the safety of rotational atherectomy of side branches jailed by stents. AB - In vitro experimental models of branch orifices jailed by various stents were created to estimate the safety and the efficacy of rotational atherectomy when rotational burrs were advanced through the struts of stents. The scaffolding structures of the stents were destroyed due to loss and deflection of the struts, and the size of ablated stent-particles differed: the maximal size was 1.7 mm in slotted stents, and 17.6 mm in coiled stents. Thus, there is a definite potential for ablating stents when rotational atherectomy of restenotic lesions of side branch orifices jailed by stents is performed. PMID- 10462022 TI - Improvement by 5-amino-4-imidazole carboxamide riboside of the contractile dysfunction that follows brief periods of ischemia through increases in ecto-5 nucleotidase activity and adenosine release in canine hearts. AB - 5-Amino-4-imidazole carboxamide (AICA) riboside increases adenosine release in ischemic myocardium, suggesting that AICA riboside improves contractile dysfunction. In 49 open-chest dogs, contractile function assessed by fractional shortening (FS) was observed 3 h after the onset of reperfusion following 15 min of occlusion of the left anterior descending coronary artery. During reperfusion, the treatment with AICA riboside increased adenosine concentration in the coronary venous blood (536+/-44 vs. 281+/-21 pmol/ml at 3 min of reperfusion, p<0.001) and peak coronary hyperemic flow (367+/-13 vs. 300+/-21 ml/100 g per min, p<0.001) when compared with the untreated group. FS at 3h of reperfusion increased in the AICA riboside group (21.1+/-2.3 vs. 12.8+/-0.6% in the untreated group, p<0.001). AICA riboside increased myocardial ecto-5'-nucleotidase activity. Administration of adenosine also augmented coronary hyperemic flow and increased FS to the levels of the AICA riboside group. Either 8 phenyltheophylline (an antagonist of adenosine receptors) or alpha,beta-methylene adenosine 5'-diphosphate (an inhibitor of ecto-5'-nucleotidase) completely abolished the increased coronary hyperemic flow and improvements of myocardial contractile function due to AICA riboside. Thus it was concluded that AICA riboside improves the contractile dysfunction that follows a brief period of ischemia via adenosine-dependent mechanisms. PMID- 10462024 TI - Pseudoaneurysm of the left ventricle progressing from a subepicardial aneurysm. AB - A 56-year-old man presented with an inferior myocardial infarction and a huge pseudoaneurysm below the inferior surface of the left ventricle, which had progressed from a small subepicardial aneurysm over a 6-month period. Transthoracic echocardiography, Doppler color flow images, radionuclide angiocardiography, magnetic resonance imaging and contrast ventriculography all revealed an abrupt disruption of the myocardium at the neck of the pseudoaneurysm, where the diameter of the orifice was smaller than the aneurysm itself, and abnormal blood flows from the left ventricle to the cavity through the orifice with an expansion of the cavity in systole and from the cavity to the left ventricle with the deflation of the cavity in diastole. Coronary angiography revealed 99% stenosis at the atrioventricular nodal branch of the right coronary artery. At surgery the pericardium was adherent to the aneurysmal wall and a 1.5 cm orifice between the aneurysm and the left ventricle was seen. Pathological examination revealed no myocardial elements in the aneurysmal wall. The orifice was closed and the postoperative course was uneventful. Over-intense physical activity as a construction worker was considered to be the cause of the large pseudoaneurysm developing from the subepicardial aneurysm. These findings indicate that a subepicardial aneurysm may progress to a larger pseudoaneurysm, which has a propensity to rupture, however, it can be surgically repaired. PMID- 10462023 TI - Plasma endothelin-1 elevation associated with alcohol-induced variant angina. AB - Vasospastic angina as a result of alcohol ingestion has been reported, but the mechanism of alcohol-induced coronary artery spasm is presently unknown. This report presents 2 cases of alcohol-induced variant angina (VA) with elevated levels of plasma endothelin-1 after alcohol ingestion. In case 1, the plasma endothelin-1 concentration was 3.15 pg/ml before drinking (normal <2.30 pg/ml) and increased to 4.09 pg/ml when measured 5 h after alcohol ingestion. After 2 months of abstinence, the plasma endothelin-1 concentration was 2.88 pg/ml and 6 months after abstinence, it decreased to 2.03 pg/ml (normal range). In case 2, the plasma endothelin-1 concentration was 2.44 pg/ml before drinking and increased to 4.36 pg/ml when measured 5 h after alcohol ingestion. After 2 months of abstinence, the plasma endothelin-1 concentration was 3.04 pg/ml and 6 months after abstinence, it decreased to 2.09 pg/ml (normal range). These 2 cases suggest that a relationship may exist between alcohol-induced VA and elevation in the plasma endothelin-1 concentration after alcohol ingestion. PMID- 10462025 TI - Multiple aortocaval fistulas associated with a ruptured abdominal aneurysm in a patient with Ehlers-Danlos syndrome. AB - Aortocaval fistula (ACF) is a rare complication of spontaneous abdominal aortic aneurysm (AAA) rupture, with an incidence of 2-4%. A unique case of ruptured AAA complicated by multiple aortovenous fistulas involving the inferior vena cava and left internal iliac vein is presented, and is the first published report of a patient with Ehlers-Danlos syndrome undergoing surgical treatment for an ACF. PMID- 10462026 TI - A case of intestinal obstruction following stent graft placement for an abdominal aortic aneurysm. AB - A 76-year-old male was admitted to hospital complaining of severe abdominal pain, constipation, nausea and vomiting. The patient had undergone stent graft placement of an abdominal aortic aneurysm (AAA) at another hospital 13 months prior to admission. An X-ray, computed tomography scan and barium-enema examination revealed partial obstruction of the duodenum. Stent graft placement has been reported to be a useful procedure for AAA. However, as mass effects associated with AAA cannot be excluded, several symptoms may remain postoperatively. PMID- 10462027 TI - Cryopreserved aortic homograft replacement in a patient with Takayasu's arteritis. AB - A patient with severe long-standing Takayasu's arteritis underwent successful replacement of the aortic root and ascending aorta with a cryopreserved aortic homograft. Her postoperative course was uneventful and echocardiography demonstrated evidence of neither aortic regurgitation nor graft detachment more than 2 years after the operation. Magnetic resonance image demonstrated no signs of graft enlargement. PMID- 10462028 TI - Biopsy-proven cardiomyopathy in heterozygous Fabry's disease. AB - A 23-year-old woman with heterozygous Fabry's disease who had acroparesthesia was admitted to hospital for precise examination of the disease before childbearing. She had no cardiac-related symptoms and no abnormality on physical examination. The alpha-galactosidase A activity in her leukocytes was present, but lower than normal. However, the endomyocardial biopsy showed specific changes for Fabry's disease. As Fabry's disease is a rare X-linked recessive inborn error of glycosphingolipid metabolism, heterozygous females are usually asymptomatic, but rarely can be affected as severely as hemizygous males. This is an isolated case of heterozygous Fabry's disease in a female in whom cardiac involvement was detected by endomyocardial biopsy, although she had no cardiac abnormality on physiological examinations. In conclusion, endomyocardial biopsy is useful for evaluation of the cardiac involvement of Fabry's disease even in an asymptomatic case. PMID- 10462029 TI - Determination of the cross-linked residues in homo-dimerization of S19 ribosomal protein concomitant with exhibition of monocyte chemotactic activity. AB - When S19 ribosomal protein molecules are intermolecularly cross-linked by a transglutaminase-catalyzed reaction, the monocyte chemotactic activity is newly expressed. Heparin, at a concentration of 1 U/ml, greatly augmented the cross linking reaction. This augmentation was due to binding affinity of S19 ribosomal protein to heparin. The major heparin-binding region of S19 ribosomal proteins was identified to Lys23-Lys-Ser-Gly-Lys-Leu-Lys29, using region-directed mutant proteins. The amino acid residues of S19 ribosomal protein used for the intermolecular cross-linkage were then determined by the peptide map analysis with amino acid sequencing and by the site-directed mutagenesis; Gln137 and Lys122 were used in the intermolecular cross-linkage. PMID- 10462030 TI - Analysis of the immunoglobulin heavy chain gene variable region of CD5-positive diffuse large B-cell lymphoma. AB - To clarify the cell origin of CD5+ diffuse large B-cell lymphoma (DLBCL), we analyzed and compared the variable region of the immunoglobulin heavy chain gene (VH gene) in eight cases of CD5+ DLBCL and 23 cases of other CD5+ B-cell neoplasms; 10 cases of chronic lymphocytic leukemia (CLL), one case of small lymphocytic lymphoma, one case of hairy cell leukemia, and 11 cases of mantle cell lymphoma. CD5+ DLBCL were comprised of two cases of de novo lymphoma of nodal origin, five cases of de novo lymphoma of extranodal origin, and one case of Richter transformation. Whereas all cases of mantle cell lymphoma except one showed a germ line or low mutation frequency of the rearranged VH gene, the rearranged VH genes in both CD5+ CLL and CD5+ DLBCL were heterogeneous. The degree of somatic mutation of CD5+ CLL and CD5+ DLBCL ranged between approximately 0 to 15.0% and 0.7 to 12.9%, respectively. High frequency of expression of the VH4 family in both CD5+ CLL and CD5+ DLBCL was found. Moreover, none of the three cases of CD5+ DLBCL examined exhibited intraclonal diversity. These findings may be common characteristics of the rearranged VH gene of CD5+ CLL and CD5+ DLBCL and suggested that the cell origin of CD5+ DLBCL was the same as that of CD5+ CLL. PMID- 10462031 TI - Phosphatase activity in the arterial wall after balloon injury: effect of somatostatin analog octreotide. AB - Phosphorylation of transcription factors fos/jun dimer activator protein (AP)-1 and nuclear factor-kappaB (NF-kappaB) plays a cardinal role in vascular smooth muscle cell (SMC) response to growth stimuli. Activity of protein tyrosine (PTP) and serine/threonine phosphatases (PP2A, B, and C) regulates in balance with the activity of protein kinases the level of transcription factor phosphorylation. Somatostatin analog octreotide stimulates phosphatase activity and inhibits cell growth. We examined in rats the activity of tissue phosphatases after arterial wall injury and treatment with octreotide and its effect on AP-1 and NF-kappaB phosphorylation and arterial response to injury. The activity of PTP did not change after balloon injury. Treatment of rats with PTP stimulator octreotide increased the PTP activity by 20% +/- 18% in uninjured arteries (p = 0.04 compared with control) and by 49% +/- 44% compared with injured untreated rats (p = 0.017). Treatment of rats with okadaic acid, a specific phosphatase inhibitor, prevented the octreotide-induced increase in PTP activity. PP2A activity of uninjured arteries was not affected significantly with treatment with octreotide (105% +/- 21%, p = 0.57 compared with control). After balloon injury PP2A activity was significantly reduced, 54% +/- 24% of control (p = 0.001). This reduction was prevented with treatment with octreotide, activity 88% +/- 25% of control. When rats were treated with octreotide and okadaic acid, the activity of PP2A in uninjured arteries was decreased to 65% +/- 12% of control (p = 0.03) and the injury-induced reduction was preserved, activity 54% +/- 8% of control (p = 0.001). There was no change in PP2B and C activity after balloon injury. Increased phosphatase activity with octreotide was associated with stabilization of the unphosphorylated form and reduction in nuclear binding of AP-1 and NF kappaB and was associated with reduced SMC proliferation after balloon injury. Inhibition of increased phosphatase activity with okadaic acid was associated with increased nuclear binding of AP-1 and NF-kappaB. Increased nuclear binding of AP-1 and NF-kappaB after injury was associated with increased expression of fos, jun, and p105 subunit mRNA and restored the proliferative response of SMC after balloon injury. We conclude that the activity of PP2A is decreased after arterial balloon injury which leads to increased AP-1 and NF-kappaB phosphorylation and nuclear binding and is involved in regulation of SMC proliferation. Treatment with octreotide prevented the injury-induced reduction in PP2A activity and decreased transcription factor phosphorylation and SMC proliferation. Modification of phosphatase activity is a potential regulatory mechanism of arterial wall response to injury. PMID- 10462032 TI - Extraneural organ involvement in human rabies. AB - Human rabies is a fatal encephalomyelitis. After the development of the central nervous system infection, there is centrifugal spread of the rabies virus to extraneural (systemic) organs. With histochemical staining and localization of rabies virus antigen (RVA) with immunoperoxidase staining, we have examined tissue sections of organs from 14 postmortem pediatric and adult cases of human rabies acquired in Mexico and the People's Republic of China. RVA was found in nerve plexuses in multiple organs, including the gastrointestinal tract. RVA was observed in muscle fibers of the heart, tongue, and larynx. RVA frequently was observed in the adrenal medulla with an associated inflammatory reaction. Minor salivary glands of the tongue contained RVA and major salivary glands showed RVA in plexuses, but not in either acini or ducts. Epithelial cells of the tongue and taste buds were occasionally infected. RVA was observed in hair follicles of the skin and rarely in pancreatic islets. The infection of extraneural organs was sometimes, but not always, associated with an inflammatory reaction. These findings indicate that centrifugal spread of rabies virus to extraneural organs occurs frequently in human rabies. PMID- 10462033 TI - Early release of mitochondrial cytochrome c and expression of mitochondrial epitope 7A6 with a porphyrin-derived photosensitizer: Bcl-2 and Bcl-xL overexpression do not prevent early mitochondrial events but still depress caspase activity. AB - Certain nonmetallic porphyrins have potent antitumor activity upon visible light irradiation. Treatment of HeLa cells with nanomolar amounts of the photochemo therapeutic agent verteporfin and red light mobilized caspases 2, 3, 6, 7, 8, and 9, caused degradation of specific caspase substrates, and resulted in morphologic changes consistent with apoptosis. Caspase processing was detectable by 1 hour after light irradiation. The mitochondrial 7A6 epitope, recognized by monoclonal antibody APO2.7, became accessible, and cytochrome c was detectable within the cytosolic fraction of cells treated with verteporfin immediately after light irradiation. The general caspase inhibitor benzyloxycarboyl-Val-Ala-Asp fluoromethylketone did not prevent 7A6 expression produced by photosensitization at peptide concentrations which completely prevented caspase activation and cleavage of caspase-specific substrates. Enforced overexpression of Bcl-2 or Bcl xL prevented cytochrome c release and 7A6 expression produced by ultraviolet B light treatment, but did not prevent cytochrome c release or 7A6 expression elicited by verteporfin photosensitization. Bcl-2 or Bcl-xL overexpression delayed morphologic changes, depressed caspase activation, and limited substrate degradation, but did not protect against loss of viability after verteporfin photosensitization. This observation indicates that cells overexpressing Bcl-2 or Bcl-xL exhibit resistance to caspase activation even after the appearance of cytochrome c in the cytosol. Porphyrin photosensitizers are effective chemotherapeutic agents that elicit primary proapoptotic mitochondrial events even in the setting of heightened Bcl-2 or Bcl-xL expression. PMID- 10462034 TI - Cellular expression of xanthine oxidoreductase protein in normal human tissues. AB - Xanthine oxidoreductase is an important cytoplasmic source of reactive oxygen species, and has been implicated in the pathogenesis of ischemia-reperfusion damage. Because the cellular localization of this protein remains unclear, our aim was to study its distribution in fresh normal human tissue obtained at surgery. For immunohistochemical studies we purified the protein from human milk and raised a polyclonal antibody in rabbits. In the liver the protein was preferentially localized to the periportal hepatocytes and it was absent from the perivenous region. In the proximal intestine, the protein was expressed in epithelial cells and goblet cells. Lactating mammary gland acinar cells showed intense staining. Small vessel vascular endothelial cells of the intestine, mammary gland, and skeletal muscle showed immunoreactivity, but in the kidney, glomerular endothelial cells were negative. No cells in the heart, brain, or lung expressed the enzyme protein. The observed localization of the xanthine oxidoreductase protein is consistent with previously observed enzyme activities in the organs studied. The widely assumed exclusive localization to capillary endothelium obviously does not apply to humans. PMID- 10462035 TI - Effects of cigarette smoke on nitric oxide synthase expression in the rat lung. AB - To examine the effects of cigarette smoke on nitric oxide synthase (NOS) gene expression and protein production, groups of rats were exposed to smoke once only or daily and were sacrificed after 1, 2, 7, or 28 days of exposure. NOS-1, NOS-2, and NOS-3 mRNAs in whole lung were quantified using reverse transcription polymerase chain reaction (RT-PCR), and NOS protein levels were determined by Western blots. Neither NOS-1 gene expression nor protein levels changed with smoke exposure. Levels of NOS-2 expression were more than doubled in smokers at Day 1 and decreased to control values during 1 month with daily smoke exposure, while protein levels did not change. NOS-3 expression was increased approximately 35% after 2 days of smoke exposure and remained increased to 28 days, whereas protein levels were increased by approximately 60% at Day 7 and remained elevated. In situ hybridization showed that NOS-2 was diffusely expressed in the lung parenchyma, airways, and vessels, and that NOS-3 was strongly expressed in vascular endothelium. Protein distribution, as determined by immunohistochemical staining, was identical to mRNA tissue distribution, and these distributions were not changed by smoke. We conclude that smoke exposure induces a rapid but transient increase in transcription of NOS-2, and a sustained increase in transcription and translation of NOS-3; up-regulation of NOS occurs within the anatomic compartment where these genes are normally expressed. These findings indicate that cigarette smoke can directly and rapidly affect NOS expression, and thus potentially affect the function of the pulmonary vasculature. PMID- 10462036 TI - The coronary endothelium: a target for vascular endothelial growth factor. Human coronary artery endothelial cells express functional receptors for vascular endothelial growth factor in vitro and in vivo. AB - Vascular endothelial growth factor (VEGF) is an angiogenic peptide that can stimulate endothelial cell proliferation and migration in vitro and collateral development in ischemic organs in vivo. Although postulated, the expression of functional VEGF receptors in the heart has not been demonstrated yet. To prove this hypothesis and to extend the molecular basis of myocardial angiogenesis, we have characterized the expression and function of VEGF receptors in human coronary artery endothelial cells (HCAEC) and in human heart tissue. VEGF strongly induces proliferation and migration of HCAEC. These cells express transcripts of the two VEGF receptors KDR and Flt-1. Their expression levels are higher in HCAEC as compared with human umbilical vein endothelial cells. In HCAEC, VEGF stimulates phosphorylation of KDR in a concentration-dependent manner proving that KDDR is a functional receptor tyrosine kinase. Scatchard analysis demonstrated the presence of the high affinity receptor Flt-1 in HCAEC with a kd of 8 pM. Flt-1 protein could be visualized as a single band corresponding to a size of 210 kd. In addition mature KDR protein could be detected in adult human heart. Taken together, HCAEC and human heart tissue express high levels of functional VEGF receptors. These results broaden the molecular basis for understanding and manipulating VEGF-induced endothelial function and angiogenesis in the coronary circulation. PMID- 10462037 TI - Suppression of telomerase activity as an indicator of drug-induced cytotoxicity against cancer cells: in vitro studies with fresh human tumor samples. AB - Telomerase is a ribonucleoprotein complex with reverse-transcriptase activity responsible for telomere reconstitution. High telomerase activity was found in cancer cells, but not in differentiated homologous nonmalignant tissues. We demonstrated previously that the disappearance of telomerase activity is a reliable marker of tumor cell killing in human cancer cell lines. We have investigated the possibility of evaluating chemosensitivity of neoplastic cells of different origin [ovary, lung, breast, gastrointestinal, skin (melanoma)] obtained from cancer patients, by measuring residual telomerase activity after drug treatment in vitro. Using the classical telomeric repeat amplification protocol ("TRAP") assay based on polymerase chain reaction, we examined telomerase activity of untreated or drug-treated tumor cell suspensions, derived from the processing of surgical specimens. Feasibility and reproducibility of the assay were evaluated according to various parameters, including drug concentration, time of in vitro culture, and type of tumor. The results indicated that the assay is highly sensitive and reproducible, and can be performed using surgical specimens in a reasonable percentage of cases, ranging from 40% (breast cancer) to 100% (ovarian cancer). Moreover, the assay provides comparable results using a wide range of tumor cells, and the presence of normal cells does not interfere with the results. Prolonged tumor cell culture is not required because the assay can be completed within 24 to 72 hours after sample collection. In conclusion, the present investigation provides the technical bases for future studies to evaluate whether this assay would be able to predict patient's response to antitumor agents. PMID- 10462038 TI - Insulin-like growth factor-II expression is down-regulated in TrkA-transfected SK N-AS neuroblastoma cells. AB - Expression level of trkA tyrosine kinase receptor for nerve growth factor is a major prognostic determinant of neuroblastoma, suggesting that defective trkA mediated signaling is responsible for the tumorigenesis of this childhood malignancy. We investigated the biologic effect of trkA, with special reference to its effect on insulin-like growth factor-II (IGF-II) expression, in SK-N-AS human neuroblastoma cells transfected with human trkA cDNA. Nerve growth factor treatment of trkA-transfected cells promoted growth and changed the morphologic phenotype into a substrate-adherent, flatter phenotype (S-type), and down regulated the mRNA expression of IGF-II. The effects on both growth and the morphologic differentiation of SK-N-AS cells differed significantly from those of previous studies, and implied that trkA effects can be diverse, depending on the phenotype of the individual neuroblastoma cells. Immunohistochemical screening of trkA and IGF-II expression in adrenal neuroblastomas (n = 25) also favored the nonoverlapping pattern of trkA and IGF-II expression (p < 0.05). Because IGF-II is believed to play a significant role in the tumorigenesis of neuroblastoma, the inverse relationship between trkA and IGF-II strongly suggests that a low level of trkA can be a feature of the pathogenetic mechanism of IGF-II expressing adrenal neuroblastomas. PMID- 10462039 TI - Interferon-beta inhibits activated leukocyte migration through human brain microvascular endothelial cell monolayer. AB - Perivascular leukocyte infiltration into the central nervous system is characteristic of multiple sclerosis (MS) pathology. Interferon-beta (IFN-beta) has shown efficacy in the treatment of patients with MS, but the relevant mechanisms remain incompletely understood. In this study the effects of IFN-beta on leukocyte transendothelial migration were investigated using cells relevant to MS pathogenesis, namely human brain microvascular endothelial cells (HB-MVEC). Activated, but not resting leukocytes exhibited a high transendothelial migration capacity. HB-MVEC prestimulated with tumor necrosis factor (TNF) and IFN-gamma significantly promoted leukocyte transendothelial migration. IFN-beta inhibited the activated leukocyte transendothelial migration on TNF/IFN-gamma-activated HB MVEC in a dose-dependent manner. A matrix metalloproteinase (MMP) inhibitor and monoclonal antibodies to lymphocyte function antigen-1 (LFA-1) or intercellular adhesion molecule-1 (ICAM-1), but not to very late antigen-4 or to vascular cell adhesion molecule-1 significantly inhibited the transendothelial migration of stimulated leukocytes, suggesting that this phenomenon involves the LFA-1/ICAM-1 interaction and MMP. However IFN-beta did not interfere with the binding of leukocytes to HB-MVEC unless IFN-beta was preincubated with leukocytes or added to HB-MVEC at the time of stimulation. Furthermore IFN-beta did not modulate the expression of adhesion molecules on either stimulated leukocytes or activated HB MVEC, but partially reduced TNF and interleukin-1 production from stimulated leukocytes during coculture with HB-MVEC. Interestingly, in the presence of IFN beta, a significant down-regulation of MMP-9 release from stimulated leukocytes was found, especially for the activated form of MMP-9. These results indicate that inhibition of leukocyte transendothelial migration is an important mechanism accounting for the beneficial effects of IFN-beta in the treatment MS patients. PMID- 10462040 TI - Redox-dependent regulation of interleukin-8 by tumor necrosis factor-alpha in lung epithelial cells. AB - Increasing evidence supports a major role for interleukin-8 (IL-8), a potent neutrophil chemoattractant, in the chronic progression of inflammatory lung diseases. The present studies were designed to characterize the molecular events involved in IL-8 induction in pulmonary epithelial cells in response to tumor necrosis factor-alpha (TNF-alpha). IL-8 induction by TNF-alpha was redox sensitive, as indicated by electron spin resonance analysis and inhibition with membrane permeable hydroxyl scavengers. Furthermore using cell transfection and mobility shift assays, it was found that transcriptional activation of the IL-8 gene required TNF-alpha-induced activation and binding of nuclear factor-kappaB (NF-kappaB)- and NF-IL-6, nuclear transcription factors to regulatory elements in the IL-8 promoter. Activation of the IL-8 promoter by these transcription factors was also redox-sensitive. This response was mediated through the TNF-R1 receptor (p55), and not the TNF-R2 (p75) receptor, although both receptors can be found on pulmonary epithelial cells. Taken together these studies indicate that TNF-alpha induced redox changes in lung epithelial cells are responsible for the transcriptional activation of IL-8 and that coordinate activation of NF-kappaB and NF-IL-6 mediate the response. PMID- 10462041 TI - A capping domain for LRR protein interaction modules. AB - Leucine-rich repeats (LRR) are protein interaction modules which are present in a large number of proteins with diverse functions. We describe here a novel motif (16-19 residues) downstream of the last, incomplete, LRR in a subfamily of LRR proteins. In the U2A' spliceosomal protein, this motif is folded into a cap that shields the hydrophobic core of the LRRs from the solvent. Modelling of the LRR cap in the imidazoline-1 candidate receptor, using the known structure of U2A' as template, showed a conservation of the basic structural features. PMID- 10462042 TI - The dermaseptin precursors: a protein family with a common preproregion and a variable C-terminal antimicrobial domain. AB - Preprodermaseptins are a group of antimicrobial peptide precursors found in the skin of a variety of frog species. Precursors of this family have very similar N terminal preprosequences followed by markedly different C-terminal domains that correspond to mature antimicrobial peptides. Some of these peptides are 24-34 amino acids long and form well-behaved amphipathic alpha-helices, others are disulfide-linked peptides of 20-46 residues, still others, highly hydrophobic, are the smallest antimicrobial peptides known so far being only 10-13 residues in length. All these peptides are broad-spectrum microbicides that kill many bacteria, protozoa, yeasts and fungi by destroying or permeating the microbial membrane. In frogs belonging to the genus Phyllomedusinae, preprodermaseptins encoded peptides also include dermorphins and deltorphins, D-amino acid containing heptapeptides which are very potent and specific agonists of the mu- or delta-opioid receptors. The remarkable similarity between preproregions of precursors that give rise to peptides with very different primary structures, conformations and activities suggests that the corresponding genes originate from a common ancestor. The high conservation of the precursor prepropart indicates that this region must have an important function. PMID- 10462043 TI - TOP mRNAs are translationally inhibited by a titratable repressor in both wheat germ extract and reticulocyte lysate. AB - Vertebrate TOP mRNAs contain a 5' terminal oligopyrimidine tract (5' TOP), which is subject to selective translational repression in non-growing cells or in cell free translation systems. In the present study, we monitored in vitro the effect of increasing amounts of a 16 nucleotides long oligoribonucleotide representing the 5' terminus of mouse ribosomal protein S16 mRNA on the translation of TOP and non-TOP mRNAs. Our results demonstrate that the wild-type sequence (but not its mutant counterparts) derepresses the translation of mRNAs containing 5' TOP motifs, but failed to stimulate the translation of non-TOP mRNAs, even if the latter differed only by a single nucleotide from their 5' TOP-containing counterparts. Similar results have been obtained with both wheat germ extract and rabbit reticulocyte lysate. It appears, therefore, that translational repression of TOP mRNAs is achieved in vitro by the accumulation of a titratable repressor rather than by the loss of an activator and that this repressor recognizes multiple TOP mRNAs with a diverse set of 5' TOP motifs. PMID- 10462044 TI - A FTIR spectroscopy evidence of the interactions between wheat germ agglutinin and N-acetylglucosamine residues. AB - Wheat germ agglutinin (WGA), a lectin binding a N-acetyl-D-neuraminic acid (NeuNAc) and/or N-acetyl-D-glucosamine (GlcNAc) group, was studied by Fourier transform infrared (FTIR) spectroscopy. Deconvolution of the FTIR spectrum of WGA alone indicated the presence of few alpha-helices and beta-sheets, in contrast to many other lectins. These results agree with previous WGA crystal data. The WGA conformational changes, induced by GlcNAc-bearing liposomes or GlcNAc oligomers, were studied by infrared differential spectroscopy. The GlcNAc binding to WGA resulted in a decrease of turns and alpha-helices and a concomitant appearance of beta-sheets, inducing more or less peptidic N-H deuteration. PMID- 10462045 TI - Mutations in the Ca2+ binding site of the Paracoccus denitrificans cytochrome c oxidase. AB - Recent structure determinations suggested a new binding site for a non-redox active metal ion in subunit I of cytochrome c oxidase both of mitochondrial and of bacterial origin. We analyzed the relevant metal composition of the bovine and the Paracoccus denitrificans enzyme and of bacterial site-directed mutants in several residues presumably liganding this ion. Unlike the mitochondrial enzyme where a low, substoichiometric content of Ca2+ was found, the bacterial wild-type (WT) oxidase showed a stoichiometry of one Ca per enzyme monomer. Mutants in Asp 477 (in immediate vicinity of this site) were clearly diminished in their Ca content and the isolated mutant enzyme revealed a spectral shift in the heme a visible absorption upon Ca addition, which was reversed by Na ions. This spectral behavior, largely comparable to that of the mitochondrial enzyme, was not observed for the bacterial WT oxidase. Further structure refinement revealed a tightly bound water molecule as an additional Ca2+ ligand. PMID- 10462046 TI - Structural and enzymatic characterization of human recombinant GDP-D-mannose-4,6 dehydratase. AB - GDP-D-mannose-4,6-dehydratase (GMD) is the key enzyme in the 'de novo' pathway of GDP-L-fucose biosynthesis. The reported cDNA sequences for human GMD predict three forms of different length, whose 'in vivo' occurrence and molecular properties are completely undefined. Here, we report the expression in Escherichia coli and the properties of each native recombinant GMD form. Only the 42 kDa long GMD (L-GMD) and the 40.2 kDa (M-GMD) forms were recovered as soluble functional proteins, while the 38.7 kDa form, short GMD (S-GMD), lacking an N terminal domain critical for dinucleotide binding, was inactive and formed a precipitate. Both L-GMD and M-GMD are homodimers and contain 1 mol of tightly bound NADP+. Their kinetic properties (Km, Kcat) are apparently identical and both forms are non-competitively feedback-inhibited by GDP-L-fucose to a similar extent. M-GMD is the predominant enzyme form expressed in several human cell lines. These data seem to suggest that modulation of the 'de novo' pathway of GDP L-fucose biosynthesis involves mechanisms other than differential 'in vivo' expression of GMD forms. PMID- 10462047 TI - TGF-beta inhibits lipopolysaccharide-stimulated activity of c-Jun N-terminal kinase in mouse macrophages. AB - Transforming growth factor-beta (TGF-beta) is a potent anti-inflammatory cytokine. Although this cytokine inhibits lipopolysaccharide (LPS)-mediated septic shock, the molecular mechanism of TGF-beta is not well known. Since recent studies showed that c-Jun N-terminal kinase (JNK), one of the mitogen-activated protein kinases, plays an important role in LPS signalling, we focused here on the inhibitory action of TGF-beta1 on LPS-stimulated JNK activity in mouse macrophages. TGF-beta1 inhibited LPS-stimulation of phosphorylated JNK1 and JNK2 and consequently of JNK activity in the cells. This JNK activity resulted in a decreased level of phosphorylated c-Jun protein. Using Western blotting, we also observed TGF-beta1 inhibition of newly synthesized c-Jun protein in LPS stimulated cells. These results demonstrate that TGF-beta1 inhibits LPS stimulated JNK activity in mouse macrophages. Also, our present study suggests a possible inhibitory mechanism of TGF-beta in signalling of LPS-induced inflammatory responses. PMID- 10462048 TI - Caspases-3 and -7 are activated in goniothalamin-induced apoptosis in human Jurkat T-cells. AB - Goniothalamin, a plant styrylpyrone derivative isolated from Goniothalamus andersonii, induced apoptosis in Jurkat T-cells as assessed by the externalisation of phosphatidylserine. Immunoblotting showed processing of caspases-3 and -7 with the appearance of their catalytically active large subunits of 17 and 19 kDa, respectively. Activation of these caspases was further evidenced by detection of poly(ADP-ribose) polymerase cleavage (PARP). Pre treatment with the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone (Z-VAD.FMK) blocked apoptosis and the resultant cleavage of these caspases and PARP. Our results demonstrate that activation of at least two effector caspases is a key feature of goniothalamin-induced apoptosis in Jurkat T-cells. PMID- 10462049 TI - ADP-ribosylation factor (ARF)-like 4, 6, and 7 represent a subgroup of the ARF family characterization by rapid nucleotide exchange and a nuclear localization signal. AB - The novel ARF-like GTPase ARL7 is a close relative of ARL4 and ARL6 (71% and 59%) identical amino acids). A striking characteristic of these GTPases is their basic C-terminus which, when fused to the C-terminus of green fluorescent protein (GFP), targets the constructs to the nucleus of transfected COS-7 cells. Full length ARL4 was detected in both nuclear and extranuclear compartments, whereas a construct of ARL4 lacking its C-terminus was excluded from the nucleus. Nucleotide exchange rates of recombinant ARL4, ARL6 and ARL7 were similar and appeared considerably higher than those of other members of the ARF family (ARF1, ARP). It is concluded that ARL4, ARL6 and ARL7 form a subgroup within the ARF family with similar, possibly nuclear, function. PMID- 10462050 TI - Identification of a multidrug resistance-like system in Tetrahymena pyriformis: evidence for a new detoxication mechanism in freshwater ciliates. AB - The freshwater ciliate Tetrahymena pyriformis is an ubiquitous organism that is present in all aquatic ecosystems. This protozoan showed a clear resistance against some polycyclic aromatic hydrocarbons which can be attributed to an efflux pump probably of the multidrug resistance (MDR) type. Immunocytochemical detection showed a positive stain of ciliate cells using the monoclonal antibodies 4E3, raised against P-glycoprotein (P-gp). The kinetics of P-gp expression were studied for control cultures and cultures treated with 15 microM benzo(a)pyrene. Western blot analysis using the Ab1, anti-P-gp polyclonal antibodies indicates the presence of two bands of 66 and 96 kDa of which the intensity increased with time in benzo(a)pyrene-treated ciliates. Uptake experiments with target compounds for the MDR pump, namely adriamycin, rhodamine 123 and two polycyclic aromatic hydrocarbons, benzo(a)pyrene and 7,12 dimethylbenzanthracene, were carried out by flow cytometry, in the presence or absence of cyclosporin (an inhibitor of the multidrug resistant pump). The data indicate that the accumulation of these compounds by ciliate cells is significantly enhanced in the presence of cyclosporin. This suggests that Tetrahymena is provided with a P-gp-like system that is functionally active in a way similar to that of the mammalian P-gp. PMID- 10462051 TI - AlphaCaMKII binding to the C-terminal tail of NMDA receptor subunit NR2A and its modulation by autophosphorylation. AB - Ca2+/calmodulin-dependent protein kinase II (CaMKII), a multifunctional, widely distributed enzyme, is enriched in post-synaptic densities (PSDs). Here, we demonstrate that CaMKII binds to a discrete C-terminal region of the NR2A subunit of NMDA receptors and promotes the phosphorylation of a Ser residue of this NMDA receptor subunit. Glutathione S-transferase (GST)-NR2A(1349-1464) binds native CaMKII from solubilised hippocampal PSDs in 'pull-out' and overlay experiments and this binding is competed by recombinant alphaCaMKII(1-315). The longer GST NR2A(1244-1464), although containing the CaMKII phosphosite Ser-1289, binds the kinase with a lower efficacy. CaMKII association to NR2A(1349-1464) is positively modulated by kinase autophosphorylation in the presence of Ca2+/calmodulin. These data provide direct evidence for a mechanism modulating the synaptic strength. PMID- 10462052 TI - Cloning, tissue distribution, subcellular localization and overexpression of murine histidine-rich Ca2+ binding protein. AB - The histidine-rich Ca2+ binding protein (HRC) resides in the sarcoplasmic reticulum of muscle and binds Ca2+. Since Ca2+ concentrations can regulate gene expression via calcineurin, the mouse homologue of HRC (mHRC) was isolated and characterized. mHRC was detected in muscle progenitor cells, in primary clonal thymic tumors and a tumor cell line, suggesting a broader role for mHRC than in Ca2+ storage during muscle contraction. mHRC was present in the perinuclear region of myoblasts. To examine if it can regulate gene expression, mHRC was overexpressed in cells differentiating into cardiac and skeletal muscle. mHRC had no effect on cardiogenesis or myogenesis. Therefore, if mHRC plays a role in the regulation of gene expression during cellular differentiation, it does not appear to be either rate-limiting or inhibitory. PMID- 10462053 TI - Identification of residues involved in v-Src substrate recognition by site directed mutagenesis. AB - To study the role of the catalytic domain in v-Src substrate specificity, we engineered three site-directed mutants (Leu-472 to Tyr or Trp and Thr-429 to Met). The mutant forms of Src were expressed in Sf9 cells and purified. We analyzed the substrate specificities of wild-type v-Src and the mutants using two series of peptides that varied at residues C-terminal to tyrosine. The peptides contained either the YMTM motif found in insulin receptor substrate-1 (IRS-1) or the YGEF motif identified from peptide library experiments to be the optimal sequence for Src. Mutations at positions Leu-472 or Thr-429 caused changes in substrate specificity at positions P+1 and P+3 (i.e. one or three residues C terminal to tyrosine). This was particularly evident in the case of the L-472W mutant, which had pronounced alterations in its preferences at the P+1 position. The results suggest that residue Leu-472 plays a role in P+1 substrate recognition by Src. We discuss the results in the light of recent work on the roles of the SH2, SH3 and catalytic domains of Src in substrate specificity. PMID- 10462054 TI - Tryptophan mediated photoreduction of disulfide bond causes unusual fluorescence behaviour of Fusarium solani pisi cutinase. AB - The fluorescence signal of the single tryptophan residue (Trp69) of Fusarium solani pisi cutinase is highly quenched. However, prolonged irradiation of the enzyme in the tryptophan absorption band causes an increase of the tryptophan fluorescence quantum yield by an order of magnitude. By using a combination of NMR spectroscopy and chemical detection of free thiol groups with a sulfhydryl reagent we could unambiguously show that the unusual fluorescence behaviour of Trp69 in cutinase is caused by the breaking of the disulfide bond between Cys31 and Cys109 upon irradiation, while the amide-aromatic hydrogen bond between Ala32 and Trp69 remains intact. This is the first example of tryptophan mediated photoreduction of a disulfide bond in proteins. PMID- 10462055 TI - Cytochrome P450 destruction by quinones: comparison of effects in rat and human liver microsomes. AB - Exposure to benzene was recently reported to lower the cytochrome P450 (CYP) content in phenobarbital-pretreated rats in vivo (Gut et al., Environ. Health Perspect. 104 (1996) 1211-1218). This study followed the ability of quinonic benzene metabolites (catechol, hydroquinone, and benzoquinone) to destroy CYP in liver microsomes from rats pretreated with various inducers and in human liver microsomes. Sensitivity of CYP isoforms to destruction was revealed and the interspecies differences assessed. The spectrophotometric evaluations of the total CYP content, assay of CYP marker activities, and electrophoresis with immunoblotting after incubation of microsomes with quinones revealed that: (1) rat liver CYP activities markedly differed in sensitivity to quinone-mediated destruction in vitro, CYP 1A and 3A being the most sensitive isoforms; (2) differences in OH radicals formation and lipid peroxidation among microsomes from rats pretreated with various CYP inducers were also observed; (3) semiquinone radical formation, OH radical production, and induction of lipid peroxidation did not contribute significantly to CYP destruction by quinones; (4) the main mechanism of CYP destruction is covalent binding of the oxidized quinone form to protein and heme moieties of CYP; (5) quinones, mainly benzoquinone, destroy human CYP isoforms to a much greater extent than rat enzymes and thus humans may be much more susceptible to the deleterious effect of benzene metabolism. In conclusion, it is suggested that CYP destruction may be another consequence of benzene exposure and should be taken into consideration when evaluations of possible health risks are performed. PMID- 10462056 TI - Binding of the Promen fluorescent probe to human serum albumin: a fluorescence spectroscopic study. AB - The binding of Promen (6-propionyl-2-methoxynapthalene) to human serum albumin (HSA) was measured by fluorescence spectroscopy, finding only one class of binding sites on the protein. Hydrophobic interactions play an important role to stabilize the complex. Attempts were made to characterize its binding site using as competitors warfarin, phenylbutazone and diazepam, which bind in a specific site or region on the HSA. Fluorescence polarization measurements and spectrofluorimetric results suggest that diazepam and Promen bind at different but interacting binding sites on the HSA. The changes in the fluorescence emission of the bound Promen in the presence of these drugs, allow to use Promen to detect unspecific interactions with the site II on the HSA. PMID- 10462057 TI - Induction of apoptosis by remoxipride metabolites in HL60 and CD34+/CD19- human bone marrow progenitor cells: potential relevance to remoxipride-induced aplastic anemia. AB - The antipsychotic agent, remoxipride [(S)-(-)-3-bromo-N-[(1-ethyl-2 pyrrolidinyl)methyl]-2,6-dimethoxybenz amide] has been associated with acquired aplastic anemia. We have examined the ability of remoxipride, three pyrrolidine ring metabolites and five aromatic ring metabolites of the parent compound to induce apoptosis in HL60 cells and human bone marrow progenitor (HBMP) cells. Cells were treated for 0-24 h with each compound (0-200 microM). Apoptosis was assessed by fluorescence microscopy in Hoechst 33342- and propidium iodide stained cell samples. Results were confirmed by determination of internucleosomal DNA fragmentation using gel electrophoresis for HL60 cell samples and terminal deoxynucleotidyl transferase assay in HBMP cells. The catechol and hydroquinone metabolites, NCQ436 and NCQ344, induced apoptosis in HL60 and HBMP cells in a time- and concentration dependent manner, while the phenols, NCR181, FLA873, and FLA797, and the derivatives formed by oxidation of the pyrrolidine ring, FLA838, NCM001, and NCL118, had no effect. No necrosis was observed in cells treated with NCQ436 but NCQ344 had a biphasic effect in both cell types, inducing apoptosis at lower concentrations and necrosis at higher concentrations. These data show that the catechol and hydroquinone metabolites of remoxipride have direct toxic effects in HL60 and HBMP cells, leading to apoptosis, while the phenol metabolites were inactive. Similarly, benzene-derived catechol and hydroquinone, but not phenol, induce apoptosis in HBMP cells [Moran et al., Mol. Pharmacol., 50 (1996) 610-615]. We propose that remoxipride and benzene may induce aplastic anemia via production of similar reactive metabolites and that the ability of NCQ436 and NCQ344 to induce apoptosis in HBMP cells may contribute to the mechanism underlying acquired aplastic anemia that has been associated with remoxipride. PMID- 10462058 TI - The events relating to lanthanide ions enhanced permeability of human erythrocyte membrane: binding, conformational change, phase transition, perforation and ion transport. AB - The binding and uptake of Gd3+ ions by human erythrocytes in vitro were studied by determining the Gd contents in membrane and in cytosol by means of particle induced X-ray emission (PIXE) spectrometry. Results obtained from varied incubation time revealed that the Gd3+ ions bind to the membrane proteins and lipids at first. Gd3+ binding to the membrane lipids and proteins lasts 0 approximately 20 and 20 approximately 100 ms respectively, as shown by the stopped-flow studies. Then a fraction of Gd3+ ions diffuses through the membrane. The kinetics of Gd3+ binding indicates that the binding to phospholipids is prior to that to the membrane proteins, but a portion of the lipid-bound Gd3+ redistributed later to the proteins. PIXE studies showed that the entry of Gd3+ increased the influx of Ca2+ and Cl-. By monitoring the changes in fluorescence of proteins and that of the Ln3+, the uptake of La3+, Eu3+, Gd3+ and Tb3+ was shown to be a process comprising a series of events. Binding to the membrane molecules induces the phase transition of lipid bilayer and conformational changes and aggregation of membrane proteins. Conformational changes of the proteins were characterized by Fourier transform IR spectroscopy (FT-IR) deconvolved spectra, i.e. alpha-helix content decreases while beta-sheet increases. ESR spectra of MSL-labeled proteins reflect the aggregation state related with the conformational change. [31P]NMR spectra of membrane lipid bilayer revealed the Ln3+ ions induced hexagonal (H(II)) phase formation. Phase transition and aggregation of membrane proteins cause the formation of domain structure and perforation in the membrane. These alterations in membrane structure are responsible for the Ln3+ enhanced membrane permeability. Thus the previous Ln3+ binding will facilitate the across-membrane transport of other Ln3+ ions through the membrane. PMID- 10462059 TI - Discovery and characterization of endogenous cannabinoids. AB - The characterization of cannabinoid receptors and signal transduction mechanisms provided the impetus for the searching for endogenous ligands for this system. The result was a family of fatty acid derivatives that interact with cannabinoid receptors to varying degrees. The two ligands that have received the most attention are anandamide (AN) and 2-arachidonolyl-glycerol (Ara-Gl). They are both present in central as well as peripheral tissues. Mechanisms for the synthesis and metabolism of AN have been described. Presently, the physiological stimuli for production and release of AN are unknown. As a result, elucidation of its physiological role remains elusive. However, it seems reasonable to conclude that both AN and 2-Ara-Gl interact with cannabinoid receptors in both peripheral and central tissue to produce a wide range of effects. Administration of these ligands to laboratory animals produce effects that are quite similar to those elicited by delta9-tetrahydrocannabinol (THC), the psychoactive constituent in marijuana. Nevertheless, there are some pharmacological differences between the plant-derived THC and the endogenous cannabinoids that could be due to either pharmadynamic or pharmacokinetics dissimilarities. Extensive structure-activity relationship studies have provided some vital insights into the actions of the endogenous ligands. First and foremost, systematic structural alterations in AN have additional support that it is acting at the cannabinoid receptors in a fashion similar to that of THC. Development of metabolically stable analogs of AN, as well as those with greater receptor affinity, have helped substantiate AN and THC similarities. Nevertheless, pharmacological differences remain between the endogenous and exogenous ligands. Whether these differences are due to the nature of their interaction with the cannabinoid receptors, activation of unique signaling pathways, interactions with non-cannabinoid receptors, or pharmacokinetic considerations remain to be resolved. PMID- 10462060 TI - Evidence for the presence of CB1 cannabinoid receptors on peripheral neurones and for the existence of neuronal non-CB1 cannabinoid receptors. AB - The discovery of CB1 and CB2 receptors and of endogenous agonists for these receptors has sparked renewed interest in the therapeutic potential of cannabinoids. This has led to a need for strategies that will provide a better separation of wanted from unwanted effects, particularly for CB1 receptor agonists. Possible strategies are to target CB1 receptors present on neurones outside the central nervous system or novel types or subtypes of neuronal cannabinoid receptor. This paper reviews evidence for the presence of CB1 receptors on peripheral neurones and for the existence of neuronal non-CB1 cannabinoid receptors. PMID- 10462061 TI - Structure-activity relationships of anandamide, an endogenous cannabinoid ligand. AB - Identification of arachidonylethanolamide (anandamide) as an endogenous cannabinoid is one of the most important developments in cannabinoid research in recent years. In a relatively short period of time thereafter, pharmacological and biochemical studies have confirmed initial speculations that anandamide is a neuromodulator and significantly advanced our understanding of cannabinoid biochemistry. Moreover, the discovery of anandamide has led to the identification of two heretofore unknown proteins associated with cannabinoid physiology: 1) Anandamide Amidohydrolase (AAH), an enzyme responsible for the hydrolytic breakdown of anandamide and 2) the Anandamide Transporter (ANT), a carrier protein involved in the transport of anandamide across the cell membrane. Evidence obtained so far suggests that these two proteins, in combination, are responsible for the termination of the biological actions of anandamide. Also, the discovery of anandamide has revealed a novel class of more selective cannabimimetic agents possessing a somewhat different pharmacological profile of potential therapeutic value. A number of such analogs have now been reported many of which possess markedly improved cannabinoid receptor affinity and metabolic stability compared to those of the parent ligand. Generally, anandamide and all known analogs exhibit significant selectivity for the CB1 receptor and modest to very low affinity for CB2. For this reason, this group of compounds can be considered as CB1 ligands. The purpose of this review is to summarize the structure-activity relationships (SAR) of anandamide for the CB1 cannabinoid receptor and to define the structural requirements for the substrates and the inhibitors of anandamide amidohydrolase and the anandamide transporter. PMID- 10462062 TI - Signal transduction of eicosanoid CB1 receptor ligands. AB - The eicosanoid ligand, arachidonylethanolamide (anandamide), interacts with the CB1 cannabinoid receptor in the brain to signal its response. Pharmacophoric points of interaction between this agonist and the receptor have been proposed based upon structure-activity relationship studies of ligand binding to the receptor. Three dimensional quantitative structure-activity relationship (3D QSAR) models have been constructed based upon the corresponding pharmacophoric points predicted for cannabinoid ligands delta9-tetrahydrocannabinol and 9-nor 9beta-hydroxyhexa-hydrocannabinol. A novel data set has been used to test the statistical validity of these models. Once the ligand interacts with the CB1 receptor, signal transduction occurs via G-proteins of the Gi/o family which are shown to be associated with the receptor. Evidence suggests that the juxtamembrane region of the C-terminal of the CB1 receptor is critical for activation of these G-proteins. PMID- 10462063 TI - Cannabinoid receptor interactions with the antagonists SR 141716A and SR 144528. AB - The G protein-coupled cannabinoid receptor subtypes CB1 and CB2 have been cloned from several species. The CB1 receptor is highly conserved across species, whereas the CB2 receptor shows considerable cross-species variations. The two human receptors share only 44% overall identity, ranging from 35% to 82% in the transmembrane regions. Despite this structural disparity, the most potent cannabinoid agonists currently available are largely undiscriminating and are therefore unsatisfactory tools for investigating the architecture of ligand binding sites. However, the availability of two highly specific antagonists, SR 141716A for the CB1 receptor and SR 144528 for the CB2 receptor, has allowed us to adopt a systematic approach to defining their respective binding sites through the use of chimeric CB1 receptor/CB2 receptor constructs, coupled with site directed mutagenesis. We identified the region encompassed by the fourth and fifth transmembrane helices as being critical for antagonist specificity. Both the wild type human receptors overexpressed in heterologous systems are autoactivated; SR 141716A and SR 144528 exhibit classical inverse agonist properties with their respective target receptors. In addition, through its interaction with the CB1 receptor SR 141716A blocks the Gi protein-mediated activation of mitogen-activated protein kinase stimulated by insulin or insulin like growth factor I. An in-depth analysis of this discovery has led to a modified three-state model for the CB1 receptor, one of which implicates the SR 141716A-mediated sequestration of Gi proteins, with the result that the growth factor-stimulated intracellular pathways are effectively impeded. PMID- 10462064 TI - Presence and functional regulation of cannabinoid receptors in immune cells. AB - In the last 30 years studies on drug-abusing humans and animals injected with cannabinoids, as well as in vitro models employing immune cell cultures, have demonstrated that marijuana and cannabinoids are immunomodulators. Both types of cannabinoid receptors, CB1 and CB2, have been found in immune cells, suggesting they are important in mediating the effects of cannabinoids on the immune system. This article reviews the data on the function and distribution of cannabinoid receptors in the immune system and their involvement in the immunomodulatory effect of these substances. PMID- 10462065 TI - Biosynthesis and inactivation of endocannabinoids: relevance to their proposed role as neuromodulators. AB - The two putative endogenous ligands of cannabinoid receptors, anandamide and 2 arachidonoylglycerol, are synthesized by and released from neurons in a Ca2+ dependent fashion, and re-uptaken and catabolized by both neurons and astrocytes. These biochemical features of the endocannabinoids, as well as some of their pharmacological effects in both central and peripheral nervous systems, suggest a role as neuromodulators for these metabolites. This neuromodulatory role is supported by the brain regional distribution of anandamide, its biosynthetic precursor and its major inactivating enzyme, and by the existence of possible regulatory mechanisms for the biosynthesis and inactivation of endocannabinoids, which are reviewed in this article. PMID- 10462066 TI - Effects of cannabinoids on energy metabolism. AB - The present review summarizes the recent work carried out by our group on the link between signal transduction pathways and metabolic regulation systems as affected by cannabinoids. In cells such as astrocytes and lymphocytes, which express cannabinoid receptors, physiologically relevant doses of cannabinoids induce a remarkable metabolic stimulation as determined e.g. by enhanced glucose utilization. Studies performed in astrocytes show that the cannabinoid-evoked stimulation of glucose metabolism is independent of adenylyl cyclase inhibition, and seems to rely on the cascade CB1 cannabinoid receptor --> Sphingomyelin breakdown --> Ceramide --> Raf-1 --> Mitogen-activated protein kinase (MAPK) --> Glucose utilization. A role for phosphoinositide 3'-kinase in the stimulation of glucose utilization by cannabinoids is also put forward. In addition, ceramide generated upon CB1 cannabinoid receptor activation may enhance ketone body production by astrocytes independently of MAPK. Anandamide has also been shown to exert metabolic effects in hepatocytes, cells that do not express cannabinoid receptors. The biological role of cannabinoids as modulators of metabolism is as yet unclear. PMID- 10462067 TI - The neurobiology of cannabinoid analgesia. AB - The discovery of cannabinoid receptors and their putative endogenous ligands raises questions as to the nature of the effects produced by cannabinoids on neural circuits that mediate pain and whether endogenous cannabinoids produced by the brain or in the periphery serve naturally to modulate pain. A sizable body of previous work showed that cannabinoid agonists suppress pain behavior in a variety of models of acute and chronic pain. However, at appropriate doses, cannabinoids also profoundly suppress motor behavior (see Sanudo-Pena et al., this volume), which complicates the interpretation of behavioral analgesia since a motor response is the endpoint of virtually all such studies. Studies conducted in this laboratory used biochemical and neurophysiological measures to determine whether cannabinoids suppress nociceptive neurotransmission. The results showed that cannabinoids suppress nociceptive neurotransmission at the level of the spinal cord and the thalamus. These effects are reversible, receptor mediated, selective for painful as opposed to nonpainful somatic stimuli, and track the behavioral analgesia both in time course and potency. PMID- 10462068 TI - Cannabinoids as potential new analgesics. AB - Among other pharmacological properties analgesia is one of the important features of cannabinoids with therapeutical prospects. Cannabinoids have been shown to produce antinociception in experimental animals and humans. Recently a new system of neuromodulation based upon the existence of cannabinoid receptors and their endogenous agonists has emerged. This has been proposed as another of the endogenous pain control systems. Current evidence indicate an interaction between cannabinoid and opioid systems, the latter being of known relevance in nociception. The fact that either exogenous or endogenous opioids enhanced cannabinoid-induced antinociception suggests simultaneous activation of both opioid and cannabinoid receptors by drugs as a new analgesic strategy. PMID- 10462069 TI - The neurobiology of cannabinoid dependence: sex differences and potential interactions between cannabinoid and opioid systems. AB - Cannabis is the most widely used illicit drug in many western countries. Its psychoactive ingredient, delta9-tetrahydrocannabinol (THC), produces a variety of effects in animals and humans that are probably mediated by specific cannabinoid receptors in the brain and interactions with several neurotransmitter and neuromodulator systems. For instance, recent research has revealed an important mutual functional relationship between cannabinoids and endogenous opioid systems in mediating the pharmacological and behavioral actions produced by these agents, including their reinforcing effects. Perinatal exposure to and interactions between cannabinoids and opioids might also have long-term behavioral consequences lasting into adulthood. In this work, we present preliminary evidence examining the potential effects of maternal exposure to THC on the motivational properties of morphine in male and female adult rats, as measured by an intravenous opiate self-administration paradigm. PMID- 10462070 TI - The effects of cannabinoids on the regulation of reproduction. AB - It has been shown that the main psychoactive component of marihuana, delta9 tetrahydrocannabinol (THC) has mainly inhibitory effects on the regulation of reproduction. Recently, the purification and availability of the endogenous ligand of the cannabinoid receptor, arachidonyl ethanolamide, anandamide, (ANA) and its specific long lasting antagonist, the SR 141716 (SR) provided us the opportunity to compare the effects of THC and ANA on the neuroendocrine regulation of reproduction. ANA decreases serum luteinizing hormone (LH) and prolactin (PRL) levels in rats of both sexes. It has no action on serum follicle stimulating hormone (FSH) level. When ANA was administered to pregnant rats it resulted in an increase of the duration of pregnancy and in the frequency of stillbirths. The postnatal development of hypothalamo-pituitary axis in offspring was temporarily inhibited. In conclusion, we found that exogenous and endogenous cannabinoids have only slightly different effects on the reproductive parameters. These effects may occur via the central cannabinoid receptor. It is possible that the sites of action are at both hypothalamic and pituitary levels. The results further support the view that ANA may be a central neurotransmitter or neuromodulator. PMID- 10462071 TI - Motor actions of cannabinoids in the basal ganglia output nuclei. AB - The levels of CB1 cannabinoid receptors in the basal ganglia are the highest in the brain, comparable to the levels of dopamine receptors, a major transmitter in the basal ganglia. This localization of receptors is consistent with the profound effects on motor function exerted by cannabinoids. The output nuclei of the basal ganglia, the globus pallidus (GP) and substantia nigra reticulata (SNr), apparently lack intrinsic cannabinoid receptors. Rather, the receptors are located on afferent terminals, the striatum being the major source. Cannabinoids blocked the inhibitory action of the striatal input in the SNr. Furthermore, cannabinoids blocked the excitatory effect of stimulation of the subthalamic input to the SNr revealing, along with data from in situ hybridization studies, that this input is another likely source of cannabinoid receptors to the SNr. Similar actions of cannabinoids were observed in the GP. Behavioral studies further revealed that the action of cannabinoids differs depending upon which input to the output nuclei of the basal ganglia is active. The inhibitory striatal input is quiescent and the cannabinoid action is observable only upon stimulation of the striatum, while the noticeable effect of cannabinoids under basal conditions would be on the tonically active subthalamic input. These data suggest that the recently discovered endogenous cannabinergic system exerts a major modulatory action in the basal ganglia by its ability to block both the major excitatory and inhibitory inputs to the SNr and GP. PMID- 10462072 TI - Cannabinoids, hippocampal function and memory. AB - Prior studies from this laboratory have shown that the psychoactive ingredient in marijuana, delta9-tetrahydrocannabinol (THC), interferes with short-term memory (1-3) in both delayed match and nonmatch to sample tasks (DMS/DNMS). Recent experiments have shown that other cannabinoids such as the potent CB1 receptor agonist, WIN 55,212-2 produces a delay-dependent deficit in the DNMS task at a dose range (0.10-0.50 mg/kg) well below that of delta9-THC which was blocked by the CB11 receptor antagonist SR141716A (Sanofi Inc). The effects of WIN 55,212-2 at low doses were similar to those of isolated lesions of the hippocampus, whereas high doses (0.50 mg/kg, i.p.) produced effects similar to lesions of both hippocampus and surrounding retrohippocampal areas. The low dose effect was delay dependent while the high dose introduced an additional deficit at short delays that was sensitive to both SR141716A and the GABA(B) receptor antagonist, phaclofen. Comparison of lesion vs. cannabinoid effects on DNMS performance suggests that CB1 receptors on hippocampal neurons interfere with the processing of DNMS task-specific information within a trial. CB1 receptors on hippocampal GABAergic interneurons and in retrohippocampal areas appear to influence the ability to maintain segregation of information between trials in the task. PMID- 10462073 TI - Role of endocannabinoids in brain development. AB - In addition to those functions that have been extensively addressed in this special issue, such as nociception, motor activity, neuroendocrine regulation, immune function and others, the endogenous cannabinoid system seems to play also a role in neural development. This view is based on a three-fold evidence. A first evidence emerges from neurotoxicological studies that showed that synthetic and plant-derived cannabinoids, when administered to pregnant rats, produced a variety of changes in the maturation of several neurotransmitters and their associated-behaviors in their pups, changes that were evident at different stages of brain development. A second evidence comes from studies that demonstrated the early appearance of elements of the endogenous cannabinoid system (receptors and ligands) during the brain development. The atypical location of these elements during fetal and early postnatal periods favours the notion that this system may play a role in specific molecular events related to neural development. Finally, a third evidence derives from studies using cultures of fetal glial or neuronal cells. Cannabinoid receptors are present in some of these cultured cells and their activation produced a set of cellular effects consistent with a role of this system in the process of neural development. All this likely supports that endocannabinoids, early synthesized in nervous cells, play a role in events related to development, by acting through the activation of second messenger coupled cannabinoid receptors. PMID- 10462074 TI - Dynamic regulation of the proinflammatory cytokine, interleukin-1beta: molecular biology for non-molecular biologists. AB - Interleukin-1beta (IL-1beta) is a key mediator and modulator of a wide array of physiological responses important for survival. It is created by a variety of cell types, including immune cells, glia, and neurons. It is a very potent biological molecule, acting both at the periphery as well as within the central nervous system. The production and release of IL-1beta is tightly regulated by far more complex processes than previously thought. An appreciation of this complexity is necessary for proper interpretation of apparent contradictions in the literature where different aspects of IL-1beta expression are measured. Given that many researchers are not molecular biologists by training, yet need an appreciation of the controls that regulate the function of key proteins such as IL-1beta, this review is aimed at both: (a) clarifying the multiple levels at which IL-1beta production is modulated and (b) using IL-1beta regulation to explain the dynamics of gene regulation to non-molecular biologists. Three major topics will be discussed. First, regulation of IL-1beta production will be examined at every level from extracellular signals that trigger gene activation through release of active protein into the extracellular fluid. Second, regulation of IL-1beta bioavailability and bioactivity will be discussed. This section examines the fact that even after IL-1beta is released, it may or may not be able to exert a biological action due to multiple modulatory factors. Last is the introduction of the idea that IL-1beta regulation is, at times, beyond the direct control of host; that is, when IL-1beta production becomes dysregulated by pathogens. PMID- 10462075 TI - Gene therapy in a rodent model of Parkinson's disease using differentiated C6 cells expressing a GFAP-tyrosine hydroxylase transgene. AB - Cells expressing a tyrosine hydroxylase (TH) cDNA under control of the promoter of the human glial fibrillary acidic protein (GFAP) gene were tested for therapeutic efficacy in a rat model of Parkinson's disease. The GFAP gene encodes an intermediate filament protein found almost exclusively in astrocytes. Its promoter is of interest for gene therapy as it is expressed in astrocytes throughout postnatal life and is upregulated in response to almost any damage to the central nervous system, including Parkinson's disease. We previously showed that a line of C6 rat glioma cells that expresses a GFAP-TH transgene, C6-THA, displays increased transgene activity when differentiated by forskolin treatment. Accordingly, the effects were investigated of implantation of both undifferentiated and differentiated C6-THA cells into the striatum of rats that had been lesioned with 6-hydroxydopamine. Implantation of either cell type produced significant behavioral recovery one week after transplantation, as judged by the turning response to apomorphine. At two and three weeks after transplantation, the behavioral effect of the undifferentiated cells was no longer statistically significant, whereas that for the forskolin-differentiated cells remained robust. Transgenic TH mRNA and protein could be detected in implants of both cell types, and in agreement with the behavioral results, levels were higher for the differentiated C6-THA cells than for the undifferentiated cells. These results indicate that the GFAP promoter is sufficiently active to enable production of therapeutic levels of dopamine from a GFAP-TH transgene, and suggest the use of astrocytes for gene therapy for Parkinson's disease. They also show that beneficial modifications of cells produced by treatment while in culture may be maintained following implantation. PMID- 10462076 TI - Alteration in brain proteins following occlusion of the middle cerebral artery in rat. AB - The effects of permanent focal ischemia on specific proteins of the cerebral hemisphere were studied by unilateral occlusion of the middle cerebral artery (MCAO) in rat. Brain proteins were prepared 72 h after the occlusion and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were identified by their interaction with rabbit antibodies against rat serum proteins and anti-transferrin antibodies. SDS-PAGE analysis of the proteins prepared from ischemic tissue showed significant increase in the 66 and 80 kDa components; where a marked decrease in the 260 kDa protein occurred in the ischemic and para-ischemic tissues. The 66 kDa and 80 kDa proteins stained intensely with anti-serum protein antibodies, indicating that they are related to plasma components. Moreover, the 66 kDa band had the same electrophoretic mobility as bovine serum albumin used as a standard molecular size marker. The 80 kDa band was identified as transferrin by staining with the specific antibody. Transferrin was immunolocalized in the penumbra of cerebral cortex, hippocampal CA1 region and dentate gyrus of the ischemic cerebral hemisphere. The present results suggest that alteration in the brain content of 66 kDa (albumin), 80 kDa (transferrin) and 260 kDa (unidentified) proteins may reflect early effects of focal ischemia. PMID- 10462077 TI - Sensitization of anorexia and locomotion induced by chronic administration of ephedrine in rats. AB - Repeated daily administration of the sympathomimetic agent ephedrine (EPH) leads to an augmentation (sensitization) of locomotor activity in rats. The present experiments examined the impact of repeated administration of the (-)- and (+) EPH enantiomers on feeding in rats to assess whether the anorexic activity of EPH exhibits tolerance or sensitization during chronic exposure and whether the time course of these effects follows that observed in studies of locomotion. Adult male rats were injected once daily for 12 days with either vehicle or 5, 10 or 20 mg/kg (-)-EPH or with 10 or 20 mg/kg (+)-EPH. Horizontal locomotion and diet consumption were assessed for 60 min in an activity chamber. Suppression of feeding and the induction of locomotion were augmented over the first four days of administration of either 10 mg/kg or 20 mg/kg of the (-)-EPH enantiomer. In contrast, repeated administration of 20 mg/kg (+)-EPH resulted in augmentation of appetite suppression but not locomotion. These results confirm and extend the phenomenon of locomotor and feeding sensitization for ephedrine, but suggest that these effects may differ for the two enantiomers of ephedrine. PMID- 10462078 TI - Role of SR-4987 stromal cells in the modulation of doxorubicin toxicity to in vitro granulocyte-macrophage progenitors (CFU-GM). AB - Bone marrow stromal microenvironment is essential for the maintenance of the hematopoietic stem cell renewal both by cell-cell interaction and cytokine production. However, stromal cells also exhibit drug metabolizing activities and they may accumulate the drug and successively affect hematopoietic progenitors by a retarded release. Our study investigated the role of both primary culture of murine bone marrow stroma and established stromal cells (SR-4987) in modulating the "in vitro" toxic activity of Doxorubicin (DXR) against murine granulocyte macrophage progenitors (CFU-GM). The main part of the study has been performed by a "in vitro" agar bilayer technique based on the CFU-GM assay performed over a feederlayer of stromal cells. The results suggest that bone marrow stromal cells play also an important role in decreasing the toxicity of Doxorubicin. Further SR 4987 stromal cells produce a Doxorubicin metabolite (not belonging to the series of metabolites described in literature) which is completely ineffective in inhibiting the growth of CFU-GM and the activity of topoisomerase I. Our data suggest that bone marrow stromal cells must be considered as a cell population having opposite pharmacological roles in modulating the drug toxicity on hematopoietic progenitors. In our model a mechanism of detoxification concerns the capacity of SR-4987 stromal cells to inactivate the drug. For a better prediction of drug hematotoxicity, it is very important to develop "in vitro" cell models able to discriminate between positive and negative modulation of drug toxicity that stromal cells can exert in the bone marrow microenvironment. PMID- 10462079 TI - Antitumoral effects of squamocin on parental and multidrug resistant MCF7 (human breast adenocarcinoma) cell lines. AB - The antiproliferative effects of squamocin, one of the easiest annonaceous acetogenins to obtain, were studied in the parental (MCF7-S) and the multidrug resistant (MCF7-R) human breast adenocarcinoma cell lines. Squamocin inhibited proliferation of both cell lines identically, by blocking the cell cycle in the G1-phase. This inhibition was reversible in the long term. Squamocin decreased the ATP pool in both MCF7 cell lines, but did not seem to induce apoptosis. Cytotoxic activity of adriamycin was not restored in MCF7-R Pgp expressing cells by squamocin addition. PMID- 10462080 TI - p-Fluorophenylglycine in the urine of baboons treated with HPTP, the tetrahydropyridine analog of haloperidol. AB - We report the presence of p-fluorophenylglycine (p-FPG) in the urine of six baboons treated with HPTP, the tetrahydropyridine dehydration product of haloperidol (HP). Oxidative N-dealkylation, the major metabolic pathway of HP, gives rise to 3-(4-fluorobenzoyl)propionic acid (p-FBPA). Subsequent beta oxidation of p-FBPA produces p-fluorophenylacetic acid (p-FPA). The presence of p FPA argues for the formation also of p-fluorophenylglyoxylic acid (p-FPGA) derived from beta-oxidation of p-FBPA. Plasma aminotransferases should convert p FPGA to p-FPG. The presence of p-FPG in these animals suggest the presence of phenylglycine aminotransferases in the baboon and possibly also in other primates, including the human. Reports by other authors found that treatment with alpha-phenylglycine (alpha-PG), an "unnatural" amino acid, leads to striatal dopamine (DA) depletion in rabbits--an effect explained on the basis of alpha-PG competing with DA for the neuronal vesicular storage sites. We performed in vitro DA release assays in mouse striatal synaptosomal preparations but found that neither alpha-PG nor p-FPG released any DA. It therefore remains unclear whether p-FPG may be a contributing factor to neurologic side-effects such as tardive dyskinesia (TD) found in patients after long-term HP treatment. PMID- 10462081 TI - Antioxidant potential of evening primrose oil administration in hyperlipemic rabbits. AB - The dietary intake of saturated fatty acids affects arteriosclerosis. We studied the effect of supplementation (15% wt/wt) of a hyperlipemic diet (1.33% cholesterol) with evening primrose oil (EPO) (Oenothera biennis) for 6 weeks in four groups of 10 rabbits each. At the end of this period we determined lipid peroxidation, glutathione content, and glutathione peroxidase, reductase and transferase activities in liver, brain, heart, aorta and platelets. The atherogenic diet increased tissue lipid peroxidation and decreased the protective antioxidant effect of glutathione. Dietary supplementation with EPO reduced tissue lipid peroxidation (61% in liver, 57% in brain, 42% in heart, 24% in aorta, 33% in platelets). Total glutathione was increased, especially in the aorta (90%) and platelets (200%); however, in all tissues the percentage of oxidised glutathione decreased. Evening primrose oil reduced glutathione peroxidase activity and increased the activities of glutathione reductase and transferase. We conclude that in rabbits made hyperlipemic with a diet rich in saturated fatty acids, EPO decreased tissue oxidative stress. PMID- 10462082 TI - Effects of chronic liver diseases on mitochondrial DNA transcription and replication in human liver. AB - To evaluate the effects of chronic liver diseases on mitochondrial DNA (mtDNA) transcription and replication, nuclear respiratory factor-1 (NRF-1) mRNA, mitochondrial transcription factor A (mtTFA) mRNA, a RNA component of ribonuclease (RNase) for mitochondrial RNA processing (MRP), mitochondrial cytochrome b mRNA, and mtDNA were measured in normal, chronically viral-hepatitic and cirrhotic human livers. The mRNA levels of the regulatory factors for mitochondrial gene (NRF-1 and mtTFA) and cytochrome b were significantly increased by chronic hepatitis (160, 280, and 175%, respectively) compared with those in normal livers, but were not different between cirrhotic and normal livers. On the other hand, concentrations of mtDNA and RNA component of RNase MRP were not different among normal, chronically hepatitic, and cirrhotic livers. These results suggest that either persistent hepatitis viral infection or repeated cell necrosis and regeneration in chronically hepatitic liver may be associated with increase in mtDNA transcription. PMID- 10462084 TI - Coronary effects of cyclovirobuxine D in anesthetized pigs and in isolated porcine coronary arteries. AB - The present study was undertaken in anesthetized pigs and in isolated porcine coronary arteries to determine the primary coronary effects of cyclovirobuxine D. In six pigs, the intravenous administration of 1.5 mg/kg of cyclovirobuxine D whilst preventing changes in heart rate and aortic blood pressure caused increases in left ventricular dP/dtmax and coronary blood flow which respectively averaged 10% and 23.9%. These responses were progressively augmented by graded increases in the dose of the drug (four pigs) and were not affected by blockade of cholinergic and adrenergic receptors (five pigs). Intravenous blockade of nitric oxide synthase (L-NAME, five pigs) abolished both responses, while intracoronary injection of L-NAME (five pigs) abolished only the coronary vasodilatation. In ten isolated coronary segments, cyclovirobuxine D significantly reduced the degree of potassium chloride-induced contraction. This reduction was not affected by inhibition of cyclooxygenase with indomethacin (five segments) or potassium channels blockade with glibenclamide (five segments), but it was abolished by L-NAME (five segments) or removal of endothelium (five segments). The present study showed that cyclovirobuxine D caused a primary effect of coronary vasodilatation, which involved mechanisms related to the endothelial release of nitric oxide. PMID- 10462083 TI - Role of genomic mechanisms on cAMP-dependent positive inotropism in isolated left atrium of rat. AB - It is well known that beta-adrenoceptor stimulation induces positive inotropism by cAMP-dependent phosphorylation of cardiac calcium channels. Furthermore, hypertrophy of different tissues including the heart have been related to the stimulation of these adrenoceptors via mechanisms coupled to activation of transcription and protein synthesis. Early effects of isoproterenol mediated via this pathway has also been associated to the stimulation of beta-adrenoceptors. However, the effects on the inotropism through genomic mechanisms have not yet been described. Isoproterenol (3 nM to 3 microM) induced a concentration dependent positive inotropism, in isolated left atrium of male Wistar rats electrically stimulated (0.5 Hz, 5 ms, 30-50% above the threshold voltage), which was antagonized by atenolol (1 microM) and inhibited by a protein kinase A inhibitor, (R)p-cAMPS (10 microM). The inhibitor of transcription, actinomycin D (4 microM), and the protein synthesis inhibitor, cycloheximide (35.5 microM), significantly decreased the positive inotropism induced by isoproterenol. Forskolin (0.1 to 3 microM), an activator of adenylyl cyclase, induced a concentration-dependent positive inotropism which was also inhibited by (R)p cAMPS, actinomycin D and cycloheximide. In the left atrium of rat, isoproterenol induced a positive inotropism which seems, at least in part, dependent upon intact transcription and protein synthesis, as suggested by the fact that the response was inhibited by the incubation with actinomycin D and cycloheximide. In addition, this genomic effect seems to be mediated by a cAMP-dependent mechanism. As it was inhibited by a protein kinase A inhibitor ((R)p-cAMPS) and similarly to isoproterenol, the positive inotropism induced by forskolin, which increases cytosolic cAMP, was also inhibited by actinomycin D and cycloheximide. PMID- 10462085 TI - Endogenous adenosine involved in the mediation of spinal antinociception produced by stimulating locus coeruleus. AB - The focus of this study was to investigate whether spinal adenosine is involved in mediating descending nociceptive modulation by the locus coeruleus (LC). Nociceptive evoked responses in parafascicular (PF) neurons were studied before and after electrical stimulation of the LC as well as before and after intrathecal (i.t.) administration of phentolamine (Ph) or aminophylline (Aph), an adenosine receptor antagonist, and 5'ethylcarboxamidoadenosine (NECA), an adenosine agonist. The main results were as follows: (1) the nociceptive evoked responses recorded in PF neurons were suppressed by LC stimulation; (2) pretreatment with i.t., Ph (40 nmol) reversed the LC effects, i.e., the suppressive effect of LC stimulation on the PF nociceptive evoked responses was reversed in the presence of Ph; (3) smaller doses of i.t. Aph (120 nmol) blocked only the suppressive effect produced by LC stimulation, while larger doses (240 nmol) reversed the LC stimulation, i.e., the LC stimulation exerted a facilatatory effect; and (4) i.t. application of NECA, an adenosine agonist, suppressed the nociceptive discharges in PF neurons. The results suggest that spinal adenosine may be involved in the mediation of the spinal antinociceptive effect produced by LC stimulation. PMID- 10462086 TI - Predictors of discharges against medical advice from a short-term hospital detoxification unit. AB - The primary goal of this study was to identify factors associated with patients leaving a 3-day hospital detoxification unit against medical advice (AMA). Medical records of 302 patients who were admitted for alcohol or other drug withdrawal were reviewed. Variables examined were: demographics, reported history of drug use, urine toxicology at admission, medication received during the detoxification, and admission day. Data were analyzed using a case-control design. Logistic regression was used to identify independent predictors. We found that being younger, having a shorter history of cocaine abuse, being admitted on a Friday and being an opiate dependent patient treated with clonidine only during the detoxification, were significantly associated with leaving AMA. These findings may provide information that can help clinicians identify those patients who are most at risk for leaving AMA. This will in turn allow them the opportunity to initiate preventive measures to decrease unnecessary attrition and improve utilisation of treatment resources. PMID- 10462087 TI - Studies of selected phenyltropanes at monoamine transporters. AB - 3-Phenyltropane analogues of cocaine are useful neurobiologic tools for examining mechanisms of neurotransmitter transporters and psychostimulant drugs. They are also potential substitute medications for psychostimulant abuse. In this study, 18 3-phenyltropane analogues were characterized in uptake and binding studies at dopamine (DAT), norepinephrine (NET) and serotonin (SERT) transporters from the rat, and in binding at DAT in rat, rhesus monkey, and human brain tissue. In rat brain tissue, potency in inhibiting uptake generally correlated with the potency in inhibiting binding at all three transporters suggesting that none of these compounds have antagonist properties. At the DAT, there was a significant correlation of inhibitory potencies between the rat and monkey, the monkey and human, and the rat and human transporters although some compounds showed some species difference. These findings suggest that with regard to the 3 phenyltropane series, there is generally little pharmacologic difference between DATs from the three species examined, although binding data from rat may not be a perfect predictor of uptake inhibition in human. PMID- 10462088 TI - Alcohol consumption and alcohol dependence in adults in New York State. AB - This objective of this study was to assess the differences by gender, age, race in the relationship between alcohol consumption and current alcohol dependence among adults in the general population. Logistic regression showed that, at levels of alcohol consumption below about six drinks per day, males have a higher probability of alcohol dependence than females, but that above about six drinks per day, females have a higher probability. Older adults are at less risk of dependence than younger drinkers; however, blacks and whites did not differ in their probability of dependence. Possible explanations for these results are discussed. PMID- 10462089 TI - Excessive discounting of delayed rewards in substance abusers with gambling problems. AB - This study evaluated delay discounting functions of substance abusing problem gamblers, substance abusing non-problem gamblers, and non-problem gambling/non substance abusing controls. Subjects chose between hypothetical monetary amounts available after various delays or immediately. In one condition, a USUS$1000 reward was delayed at intervals ranging from 6 h to 25 years. At each delay interval, the immediately available rewards varied from USUS$1 to USUS$999 until choices reflected indifference between the smaller immediate and larger delayed rewards. In a second condition, the delayed reward was USUS$100, and immediate rewards varied from USUS$0.10 to USUS$99.90. In all three groups, hyperbolic discounting functions provided a good fit of the data, and the smaller reward was discounted at a higher rate than the larger reward. Substance abusers discounted delayed rewards at significantly higher rates than non-substance abusing controls, and problem gambling substance abusers discounted delayed rewards at higher rates than their non-problem gambling substance abusing counterparts. Discounting rates were not associated with types or recency of drug use. These results provide further evidence of more rapid discounting of delayed rewards in substance abusers, and especially among substance abusers with a comorbid gambling problem. Rapid discounting of delayed rewards may be a feature central to impulse control and addictive disorders. PMID- 10462090 TI - Comparison of urinary 5-hydroxytryptophol, breath ethanol, and self-report for detection of recent alcohol use during outpatient treatment: a study on methadone patients. AB - This study compared urinary 5-hydroxytryptophol (5HTOL) with breath-ethanol testing as objective ways to disclose recent drinking by outpatients attending a methadone maintenance treatment clinic. Information about quantity and frequency of alcohol use was obtained by confidential self-reports. Random screening was performed on Mondays-Fridays in connection with routine clinic visits for methadone dosing. An observed urine sample for monitoring of illicit drug use and determination of 5HTOL, expressed as a ratio to 5-hydroxyindole-3-acetic acid (5HIAA), was obtained from 202 patients (59 women and 143 men), 16 of whom refused to complete the self-report and/or do a breath-ethanol test. Patients taking disulfiram or calcium carbimide for alcohol detoxification were excluded. Among the 177 subjects remaining, 47 (26.6%) reported intake of any alcohol on the previous day (range, 10-230 g ethanol; median, 40). Only four of those could be identified by a positive breath-test, while 17 showed a urinary 5HTOL/5HIAA ratio above the cutoff limit. Their alcohol consumption (median, 60 g) was significantly higher compared with those showing ratios within the reference interval (median, 35 g). The sensitivity of 5HTOL/5HIAA testing for detecting self-reported drinking in excess of 50 g ethanol was 77%. An additional nine patients who claimed abstinence still showed abnormal 5HTOL/5HIAA ratios, and so did three of the patients who refused to do a breath-ethanol test and/or complete the self-report. Altogether, 59 of 190 methadone-maintained patients (31.1%) had been drinking any alcohol on the previous day (i.e. Sunday-Thursday) according to self-report and/or urinalysis data, 29 (49.2%) of whom were identified by the urinary 5HTOL/5HIAA ratio and only four (6.8%) by utilizing breathalyzer. PMID- 10462091 TI - Depressive symptomatology and cocaine-induced pituitary-adrenal axis activation in individuals with cocaine dependence. AB - The hypothalamic-pituitary-adrenal (HPA) axis plays a role in cocaine dependence and major depressive disorder. The authors examined the correlation between baseline depressive symptomatology and pituitary-adrenal axis activation induced by acute cocaine challenge. Twelve patients with cocaine dependence were administered an iv bolus of cocaine (0.6 mg/kg) and their plasma was assayed for levels of adrenocorticotropic hormone (ACTH) and cortisol. Depressive symptomatology was assessed with total Hamilton rating scale for depression (HRSD) scores and its vegetative and cognitive superfactors. Cocaine produced a mean increase from baseline of 261% for ACTH and 73% for cortisol plasma levels. Changes in ACTH (r=0.69) and cortisol (r=0.59) were positively and significantly correlated with total HRSD scores and its vegetative, but not cognitive, factor symptom cluster. These results suggest that the HPA axis may be involved in affective disturbances associated with the use of cocaine. Implications of these data for the pathophysiology of cocaine dependence are discussed. PMID- 10462092 TI - Drug and alcohol consumption and sexual risk behaviour among young adults: results from a national survey. AB - To study the association of the consumption of alcohol and other psychoactive drugs with sexual risk behaviour for HIV infection, data from a representative sample of the Spanish population aged 18-39 years were analysed. A national household survey was carried out in 1996 using a combination of face-to-face interviews and self-administered questionnaires. The survey included 5253 subjects aged 18-39 years who provided information on alcohol and drug consumption, number of sexual partners and condom use with the steady partner and with casual partners in the 12 months before the survey. Of those surveyed, 27.4% had been drunk at least once and 20.5% had consumed drugs. Both behaviours were associated with male sex, younger age, higher educational level, being single and having had more than one sexual partner. In the logistic regression analysis adjusting for the sociodemographic variables, the greater frequency of drunkenness and cannabis use were associated with having more than one sexual partner. Regular condom use was significantly less frequent among cocaine users and more frequent among opiate users, but was not associated with the use of other drugs. Sexual risk behaviour (i.e. more than one partner and failure to use a condom regularly) was more frequent among persons who had been drunk or used cannabis or cocaine. Excessive consumption of alcohol, and cannabis and cocaine use are independently associated with sexual behaviour involving greater risk of HIV infection or transmission. PMID- 10462093 TI - Pharmacokinetic comparison of the buprenorphine sublingual liquid and tablet. AB - Buprenorphine is a mu opioid partial agonist being developed as a treatment for opioid dependence. Buprenorphine, usually administered as a sublingual liquid, is now being developed as a sublingual tablet for clinical use. The present study compared participants' plasma concentrations after daily maintenance on three buprenorphine liquid doses (2, 4 and 8 mg) and one tablet dose (8 mg). Fourteen opioid-dependent individuals (11 males, three females) participated. Plasma samples were collected over a 24-h period after at least 7 days of maintenance on each dose. Results showed that the liquid doses produced dose-related increases in plasma concentrations. The 8-mg tablet produced mean plasma concentrations significantly lower than those of the 8-mg liquid, although there was substantial individual variability. Thus, the buprenorphine tablet dose might have to be adjusted to produce plasma concentrations equivalent to those of the liquid. PMID- 10462094 TI - Methadone treatment in Spain, 1994. AB - The objective of this study was to describe the methadone treatment (MT) network in Spain, and to evaluate compliance with criteria known to influence the spread of human immunodeficiency virus (HIV). During the last 3 months of 1994, a mailed questionnaire was sent to the coordinators of all methadone treatment centers in Spain. A total of 224 centers completed the questionnaire (90% response rate). The total number of patients receiving MT in the studied centers was 13,402. Their mean age was 29 years, 79% were male, and approximately 60% were HIV positive. The rate of patients in MT varied by Region (mean: 6.7 patients in MT per 10,000 inhabitants). Although the global mean of reported daily dose of methadone was 60 mg, in 44% of the centers it was lower than that. Despite the high number of centers involved with MT in Spain, the coverage by regions is unequal. Studied centers revealed only a moderate adherence to procedures considered to be effective in HIV prevention. Given the magnitude of HIV infection in Spain, there is a clear need for improvement. PMID- 10462096 TI - Relationships between characteristics of drinking occasions and negative and positive experiences related to drinking. AB - The relationships between characteristics of drinking occasions and experiences related to drinking were studied. Interviews were conducted in 1992 with a representative sample of the Finnish population between 15 and 69 years of age. The analyses were based on 10,039 drinking occasions of 1,483 male and 1,373 female drinkers. Intake of alcohol and the frequency of drunkenness were the most important predictors of both negative and positive experiences related to drinking. Characteristics of the drinking situations contributed significantly to the explanatory models, but their predictive power was small. Public and large group drinking situations tended to increase and private and intimate situations tended to decrease the number of both negative and positive experiences related to single drinking occasions. The same situations that lead to increased positive experiences with drinking also increase the risk for negative experiences. PMID- 10462095 TI - Predicting proximal factors in cocaine relapse and near miss episodes: clinical and theoretical implications. AB - This study examined the degree of correspondence between relapse vulnerability factors assessed at intake to aftercare in 100 cocaine dependent patients and proximal factors in their first cocaine relapse and near miss episodes during a 1 year follow-up. Proximal factors in relapse and near miss episodes were also compared. Correspondence between experiences associated with prior use and experiences in the week prior to relapse and near miss episodes was generally poor. Psychiatric and family/social problem severity and coping factors at intake to aftercare predicted experiences in the week prior to near misses, and to a lesser degree, experiences in the week prior to relapse episodes. However, relapse vulnerability factors were also associated with psychiatric and family/social problem severity and mood during abstinent periods. Therefore, there was little evidence of specificity in relationships between relapse vulnerability factors and experiences prior to relapse. Proximal measures of coping, sensation seeking, positive experiences, and unpleasant affect differentiated relapses from near misses in a within-subjects analysis. PMID- 10462097 TI - Community reinforcement training for family and significant others of drug abusers: a unilateral intervention to increase treatment entry of drug users. AB - We randomly assigned 32 concerned family members and significant others (FSOs) of drug users (DUs) to a community reinforcement training intervention or a popular 12-step self-help group. We measured problems arising from the DU's behavior, social functioning of the DU and FSO, and mood of the FSO at baseline and 10 weeks later. We also monitored the FSOs' treatment attendance and treatment entry of the DUs. The treatment groups showed equal reductions from baseline to follow up in problems and improvements in social functioning and mood of the FSO. However the community reinforcement intervention was significantly better at retaining FSOs in treatment and inducing treatment entry of the DUs. PMID- 10462098 TI - Extended survival time following pseudorabies virus injection labels the suprapontine neural network controlling the bladder and urethra in the rat. AB - The central neurons that are involved in control of the urinary bladder and proximal urethra in adult male rats were identified by retrograde transport of the viral transneural tracer pseudorabies virus (PRV, Bartha strain). At 5 days post-injection, PRV-infected neurons were found in suprapontine central nervous system nuclei including ventrolateral periaqueductal gray, magnocellular division of the red nucleus, lateral hypothalamus and paraventricular nucleus, retrochiasmic region and suprachiasmatic nucleus. At days 6 and 7 PRV-infected neurons were observed in the amygdala, lateral septal nucleus, hippocampus, and frontal motor, piriform, and perirhinal cortices. These results identify the supraspinal neural networks that are involved in control of the lower urinary tract, and demonstrate the utility of long survival times to label higher-order neurons with PRV. PMID- 10462099 TI - Hypoxia- and hypercapnia-induced DC potential shifts in rat at the scalp and the skull are opposite in polarity to those at the cerebral cortex. AB - In anaesthetized and artificially ventilated rats DC (direct current) potential shifts induced by hypoxia or hypercapnia for 10 min were monitored at the surface of the skin, of the skull and of the cerebral cortex. Hypoxia was induced by decreasing the inspiratory O2 content from 20 to 10 or to 6% O2 in N2. Hypercapnia was induced by applying gas mixtures with CO2 contents from 0 to 5, 10, 20 or 30% CO2 in O2. DC potentials were recorded with non-polarizing electrodes filled with 150 mM NaCl solution. Hypercapnia evoked a negative DC shift (1.2-1.9 mV) epicranially and a large positive DC deflection (8.8-17.1 mV) epidurally. In contrast, hypoxia elicited a positive DC shift in skull recordings with a negative DC shift (2.6-3.1 mV) overshooting the baseline during recovery. DC shifts at the skin were positive, but smaller in amplitude (up to 3.1 mV). At the surface of the dura hypoxic DC shifts were negative and smaller (1.5-2.2 mV) than at the skull. The results show that a negative DC shift induced by hypercapnia recorded non-invasively from the skin or the skull is not always reflecting an increased cortical activation state and, vice versa, a positive DC shift does not always reflect a decreased state of neuronal activation. The electrogenesis of gas content-induced potentials could be due to the electrochemical diffusion of ions through the selective permeability barrier between the blood and tissue compartment. These observations are relevant for any application of non-invasive DC electroencephalography in the human cortex. PMID- 10462100 TI - Immunohistochemical evidence for the presence of calbindin containing neurones in the myenteric plexus of the guinea-pig stomach. AB - Using immunohistochemistry we studied the presence of calbindin in myenteric neurones of the guinea-pig stomach. A rabbit anti recombinant rat calbindin-D28k (CALB) stained 12, 12 and 25% of all myenteric neurones in the fundus, corpus and antrum, respectively. A rabbit anti recombinant human CALB stained 4, 4 and 16%, respectively. A mouse monoclonal antibody against the chicken intestinal CALB showed no labelling. In all regions most calbindin neurones were additionally choline acetyltransferase (ChAT) positive while only a small proportion exhibited nicotinamide adenosine dinucleatide phosphate (NADPH)-diaphorase-activity. Numerous calbindin-positive varicose nerve fibres were present within myenteric ganglia, rarely detectable in the muscle layers and virtually absent in the mucosa. This study demonstrated that a supopulation of cholinergic myenteric neurones in the stomach contain calbindin and suggested that many of these neurones fulfil interneuronal tasks. PMID- 10462101 TI - Changes in binding to muscarinic and nicotinic cholinergic receptors in the chick telencephalon, following passive avoidance learning. AB - Changes in nicotinic and muscarinic cholinergic receptors 30 min after one-trial passive avoidance training were studied in day-old chicks (Gallus domesticus), by quantitative receptor autoradiography. [3H]-alpha-bungarotoxin (alpha-BgT) and [3H]-quinuclidinyl benzilate (QNB) were used to monitor changes in 15 forebrain regions for nicotinic and muscarinic receptors, respectively. A significant increase occurred bilaterally in the quantity of bound alpha-BgT in the lobus parolfactorius, while the amount of bound QNB decreased significantly, and bilaterally, in the hippocampus, hyperstriatum ventrale, lobus parolfactorius and posterolateral telencephalon, pars dorsalis. The data support an involvement of cholinergic receptor types in the neural mechanisms underlying passive avoidance learning. PMID- 10462102 TI - L-Deprenyl stimulates the release of catecholamines in the rat medial basal hypothalamus in vivo. AB - In vivo release of catecholamines in the medial basal hypothalamus (MBH) by L deprenyl, a monoamine oxidase-B (MAO-B) inhibitor, was measured in young male Sprague-Dawley rats with stereotaxically implanted push-pull cannulae in the MBH and perfused with 0 (control), 1.5, 2.5 or 10.0 microg deprenyl in 20 microl of saline. Perfusate samples were collected at 20-min intervals and analyzed for norepinephrine (NE) and dopamine (DA) by high performance liquid chromatography (HPLC)-EC. NE release in the MBH was enhanced following perfusion with 2.5 and 10.0 microg deprenyl while DA release was augmented after infusion of 10.0 microg of deprenyl. There were no significant alterations in the release of NE and DA in the control and 1.5 microg deprenyl groups. These results suggest that deprenyl induced in vivo release of catecholamines in the MBH may be involved in the reversal of some of the reproductive aging processes. PMID- 10462103 TI - Priming stimulation in the basolateral amygdala modulates synaptic plasticity in the rat dentate gyrus. AB - We investigated the effects of basolateral amygdala (BLA) priming on long-term potentiation (LTP) in the dentate gyrus (DG). In the control animals, the induction of high-frequency stimulation (HFS) to the perforant path (PP) resulted in hippocampal LTP at all the time intervals tested. A priming stimulation to the BLA prior to the application of HFS to the PP resulted in the enhancement of the excitatory post-synaptic potential (EPSP)-LTP and population spike (PS)-LTP in the DG from 90-min post-HFS onwards. These findings suggest that the amygdala has a potential role in the modulation of some aspects of memory that are mediated by the hippocampus. PMID- 10462104 TI - Presynaptic involvement of nitric oxide in dopamine D1/D5 receptor-induced sustained enhancement of synaptic currents mediated by ionotropic glutamate receptors in the rat hippocampus. AB - Using whole-cell patch-clamp recordings, the study tested the possibility whether nitric oxide (NO) was involved in dopamine D1/D5 receptor (D1/D5r)-induced sustained enhancement of excitatory postsynaptic currents (EPSCs) mediated by ionotropic glutamate receptors in rat hippocampal slices. Activation of D1/D5r by a selective agonist, 50 microM (+/-)-6-chloro-PB hydrobromide, elicited not only a sustained enhancement but also an increased frequency of spontaneous EPSC. The D1/D5r-induced effect was associated with decreases in paired-pulse facilitation (PPF). A selective inhibitor of neuronal NO synthase (nNOS), 100 microM 7 nitroindazole monosodium salt, substantially attenuated both the magnitudes of D1/D5r-induced enhancement and PPF decrease. These results suggest that presynaptic effects mediated by NO, possibly synthesized by nNOS, are involved in D1/D5-induced sustained enhancement of synaptic currents mediated by ionotropic glutamate receptors in the hippocampus. PMID- 10462105 TI - Phosphorylated tau in human cerebrospinal fluid is a diagnostic marker for Alzheimer's disease. AB - Microtubule-associated protein tau in cerebrospinal fluid (CSF) has been proposed as a diagnostic marker for Alzheimer's disease (AD), but there is overlap between AD patients and non-AD controls. To improve the diagnostic accuracy, we measured phosphorylated tau in CSF, because phosphorylated tau accumulates as pathological paired helical filaments in neurons of the AD brain. Immunoblot showed that CSF contained a 32 kDa N-terminal fragment of tau that was partially phosphorylated on Ser199, Thr231 and Ser235. A sandwich enzyme immunoassay revealed that phosphorylated CSF-tau levels were significantly higher in AD patients than those in non-AD controls. Discrimination between the two groups was clearer in phosphorylated CSF-tau than in total CSF-tau. The data indicate that elevated phosphorylated CSF-tau level is a more specific diagnostic marker for AD. PMID- 10462106 TI - Metabotropic glutamate receptors 2 and 3 expressed by astrocytes in rat ventrobasal thalamus. AB - It has been reported that pharmacologic activation of metabotropic receptors mGluR2/3 may produce inhibition of GABAergic transmission in rat ventrobasal thalamus. We use double immunolabeling to show that: (i) mGluR2/3 is expressed in glial processes surrounding GABAergic terminals, rather than in the terminals themselves, and (ii) mR2/3 positive glial lamellae more frequently surround GABAergic terminals and ascending terminals, than cortical terminals. This suggests involvement of glia in glutamate/GABA interactions in the sensory thalamus. PMID- 10462107 TI - The effects of single and multiple episodes of theta patterned or high frequency stimulation on synaptic transmission from hippocampal area CA1 to the subiculum in rats. AB - Long-term potentiation (LTP) is a popular model for the synaptic changes that may occur during learning and memory; it involves a strengthening of synaptic response and is readily induced in the hippocampus, an area of the brain implicated in learning and memory. Previous research on LTP has focused on 'early' components of the hippocampal circuitry, that is, the dentate gyrus and areas CA1 and CA3. This paper examines the plasticity of the CA1-subiculum pathway; we extend our previous work in this area demonstrating that the projection from area CA1 to subiculum sustains theta-patterned stimulus-induced LTP in vivo. We show that this pathway remains potentiated over a long period (3 h). Furthermore, once this projection is potentiated, it seems resistant to further episodes of high-frequency stimulation. We discuss the implications of these findings for theories of hippocampal-cortical interaction during the biological consolidation of memory. PMID- 10462108 TI - Protection against acute amphetamine-induced behavior by microinjection of a group II metabotropic glutamate receptor agonist into the dorsal striatum of rats. AB - Group II metabotropic glutamate receptors (mGluRs) are distributed both pre- and postsynaptically in the striatum. By bilaterally administering a subgroup selective agonist or antagonist into the dorsal striatum of chronically cannulated rats, this study examined the role of striatal group II mGluRs in the regulation of basal and dopamine-stimulated motor behavior. Intrastriatal injection of a group II agonist, (2S,2'R,3'R)-2-(2',3' dicarboxycyclopropyl)glycine (DCG-IV, 0.01, 0.1 and 1 nmol), dose-dependently reduced basal levels of motor activity. Pretreatment of rats with intrastriatal DCG-IV at a higher dose (1 nmol), but not a lower dose (0.01 nmol), produced complete or partial blockade of hyperlocomotion induced by acute injection of amphetamine (2.5 mg/kg, i.p.) or apomorphine (1 mg/kg, s.c.), respectively. Blockade of group II mGluRs by intrastriatal injection of a group II antagonist, (RS)-alpha-methylserine-O-phosphate monophenyl ester (10 nmol), was found to: (i) induce a moderate locomotion by itself; (ii) augment amphetamine-stimulated behaviors and (iii) attenuate DCG-IV-induced reduction of basal and amphetamine stimulated motor activity. These data demonstrate that the group II mGluRs in the striatum play a significant role in the inhibitory modulation of tonic and phasic motor activity, which is most likely processed through both pre- and postsynaptic mechanisms. PMID- 10462109 TI - Memory deterioration in an oxidative stress model and its correlation with cytological changes on rat hippocampus CA1. AB - Exposure to ozone results in an increased production of free radicals which causes oxidative stress. The purpose of this study was to determine the effects of ozone exposure on memory and its correlation with the cytology of the hippocampus. Twenty-four male Wistar rats were exposed to 1 ppm (parts per million) ozone for 4 h in a closed chamber. Control group was exposed to flowing air. After ozone exposure, the rats were given long-term (24 h) memory training which consists of a passive avoidance conditioning. After that the animals were perfused and the brains were placed in the Golgi stain. The analysis consisted in counting the dendritic spines in five secondary and five tertiary dendrites of each of the 20 pyramidal neurons of hippocampus CA1 analyzed. Our results showed alterations on long-term memory and a significant reduction of dendritic spines, and provided evidence that this deterioration in memory is probably due to the reduction in spine density in the pyramidal neurons of hippocampus. PMID- 10462110 TI - No mutation in the entire coding region of the alpha-synuclein gene in pathologically confirmed cases of multiple system atrophy. AB - To determine whether mutations in the coding region of the alpha-synuclein gene are relevant in cases of multiple system atrophy (MSA), detailed nucleotide sequence analysis of the alpha-synuclein gene was performed using total RNA obtained from autopsied brain specimens of 11 pathologically confirmed cases of MSA. The brain specimens used in this study contained both gray and white matter, which were dissected from the frontal, temporal or occipital lobe. No nucleotide alterations were found in the entire coding region of the alpha-synuclein gene in any of the cases. While mutations in the regulatory or intronic regions of the gene were not ruled out, our results suggest that mutations in the coding region of the alpha-synuclein gene are unlikely to contribute to the pathogenesis of MSA. PMID- 10462111 TI - Apolipoprotein E and alpha-1-antichymotrypsin allele polymorphism in sporadic and familial Alzheimer's disease. AB - Alzheimer disease (AD) patients with both sporadic and familial forms of AD and non-demented controls were genotyped for common polymorphisms in the signal peptide for alpha-1-antichymotrypsin (ACT) gene and in two different regions of apolipoprotein E (APOE) gene. The ACT TT genotype was over-represented (P = 0.025) in patients with early onset of sporadic AD. In this patient's group ACT TT genotype conferred a significant crude odds ratio for the disease (OR = 2.09; 95% CI = 1.09-4.00, P = 0.025). After adjustment for the APOE epsilon4 and APOE 491 genotypes, logistic regression analysis confirmed that the ACT TT genotype resulted independently associated with early onset AD (adjusted OR = 2.56; 85% CI = 1.3-5.2, P = 0.009). The frequency of APOE epsilon4 allele was increased in AD, as expected (OR = 5.92, 95% CI = 3.60-9.70, P = 0.0001). On the contrary, the APOE -491 A/T genotypes were not associated with AD. No preferential association of the APOE epsilon4 allele or APOE -491 A/T genotypes with ACT A/T alleles was observed in AD. Present findings indicated that subjects with ACT TT genotype had an increased risk of developing AD and suggested that this genotype influenced the risk of an early onset of the disease by affecting the production of ACT molecules. PMID- 10462112 TI - The alpha-2 macroglobulin gene is not associated with Alzheimer's disease in a case-control sample. AB - Genetic association has recently been reported between alleles in the alpha-2 macroglobulin (A2M) gene and the occurrence of Alzheimer's disease (AD) in familial and sporadic samples. We have investigated the A2M intronic deletion polymorphism in a case-control study of 295 unrelated clinic and community-based AD cases, and compared these to a sample of 113 unrelated control individuals recruited as part of an epidemiological study. Our results show no association between A2M and AD in either case sample. Furthermore, A2M is not predictive of AD in an interactive fashion when considering APOE, race or gender. In a subset of our larger sample we have also investigated the A2M Val1000lle polymorphism, and again find no evidence for association. We conclude that there is no genetic association between A2M and AD in our case-control sample. PMID- 10462113 TI - Influence of the N-methyl-D-aspartate antagonist memantine on human motor cortex excitability. AB - The aim of our study was to investigate the effect of the N-methyl-D-aspartate (NMDA) antagonist memantine on motor excitability in humans. Seven healthy volunteers received memantine or placebo, respectively, over a period of 8 days. At day 8, transcranial magnetic stimulation (TMS) was performed using a paired pulses paradigm in order to assess intracortical inhibition and facilitation. Additionally, motor threshold and silent period duration after TMS were measured as well as M waves, F waves and peripheral silent period after electrical peripheral nerve stimulation. Intracortical inhibition was enhanced, and intracortical facilitation reduced after memantine ingestion in comparison to placebo, whereas no significant difference could be observed regarding the other neurophysiological parameters. We conclude that the NMDA receptor is involved in the regulation of excitability of intracortical interneuronal circuits. PMID- 10462114 TI - Unipolar brush cells in the cochlear nuclei of a primate (Callithrix jacchus). AB - Unipolar brush cells (UBCs) have been recognized in the cerebella of many species including primates. They have also been identified in the cochlear nuclei (CN) of many mammals, however, not in any primate. Our immunohistochemical study in the marmoset demonstrates the presence of abundant calretinin immunoreactive (CR-ir) UBCs in the cerebellum, as in other mammals. In the marmoset CN, in contrast, CR ir UBCs could hardly be identified. The reverse pattern was evident in sections immunostained for a metabotropic glutamate receptor (mGluR2/3-IR): beautifully stained UBCs were present in the CN, but the stained cerebellar structures were difficult to identify as UBCs. The present findings indicate that UBCs are present in the CN of primates, as in other mammals, but that cerebellar and CN UBCs might differ in their molecular equipment in primates. PMID- 10462115 TI - Changes with aging and ischemia in nicotinic acetylcholine receptor subunit alpha7 mRNA expression in postmortem human frontal cortex and putamen. AB - Age-related changes in nicotinic acetylcholine receptor (nAChR) subunit alpha7 messenger RNA (mRNA) expression in postmortem human frontal cortex and putamen of controls and status lacunaris patients were investigated using nonradioactive reverse transcription(RT)-PCR. In the frontal cortex of control brains, alpha7 subunit mRNA significantly decreased with age (P < 0.05). In the putamen, alpha7 subunit mRNA expression was significantly lower than that in the frontal cortex (P < 0.0001), and showed no significant correlation with age. However, in cases with status lacunaris in the putamen, alpha7 subunit mRNA expression was significantly higher compared with controls (P < 0.001). The reduction in alpha7 nAChR in the frontal cortex with age may decrease functional cholinergic synapses and cortical activity, and play a role in the cognitive impairments associated with normal aging. The functional significance of the upregulation of alpha7 nAChR mRNA in ischemic conditions remains to be determined. PMID- 10462116 TI - Magnetic resonance image-based cerebellar volumetry in healthy Korean adults. AB - The effects of age and gender on cerebellar size have not been established yet. To understand these effects, the area of cerebellar vermis and the volume of cerebellum were measured using serial magnetic resonance images of 124 Korean adults free of neurologic symptoms and signs. Cerebellar volume of male was significantly larger than that of female, although the size of vermis did not show significant gender difference. Correlation analysis revealed that cerebellar volume was not affected by aging. Regressional analysis demonstrated that female vermis had a tendency to shrink after age of 50, whereas male vermis and total cerebellar volume in both sexes were not altered with aging. The different response of vermis with aging and maintenance of cerebellum volume need to be more explored. PMID- 10462117 TI - Emperipolesis of lymphoid cells in vagal efferent neurons following an intraneural injection of ricin into the vagus nerve in rats. AB - Injection of a minute amount of the toxic lectin, Ricinus communis agglutinin-60 (RCA-60) into the vagus nerve resulted in a selective destruction of the vagal efferent neurons in the ipsilateral dorsal motor nucleus (DMN). This has elicited a massive influx of mononuclear leucocytes, notably macrophages and T lymphocytes, as detected with ED-1 and OX-19 antibodies, respectively. A small number of B-lymphocytes as identified by OX-33 antibody, were also observed in the neuropil of DMN. The influx of mononuclear leucocytes into the neuropil of DMN was by way of diapedesis, peaking in frequency at 4-6 days after the RCA administration. The infiltrated lymphocytes were closely associated with or penetrated the soma of the vagal neurons, some bearing intact axo-somatic synaptic contacts. The entrapped lymphocytes in neurons underwent degeneration and subsequently disintegrated. Macrophages and plasma cells in the neuropil did not appear to penetrate the neuronal soma. It is concluded that emperipolesis of lymphocytes, presumably cytotoxic T-cells, in RCA-poisoned neurons may represent a form of effector-target cell contact leading to cytotoxicity. In doing so, however, the invading lymphocytes were destroyed by the contents of RCA picked up by the neurons. The absence of macrophages and plasma cells in the RCA-poisoned neurons suggests the cellular specificity of emperipolesis. PMID- 10462118 TI - Activation of respiratory afferents by resistive loaded breathing modifies somatosensory evoked potentials to median nerve stimulation in humans. AB - The cortical projections of respiratory afferents (vagus and respiratory muscle nerves) are well documented in humans. It is also shown that their activation during loaded breathing modifies the perception of tactile sensation as well as the motor drive to skeletal muscles. The effects of expiratory or inspiratory loaded breathing on somatosensory evoked potentials (SEPs) elicited by median nerve stimulation were studied in eight healthy subjects. No significant changes occurred in latencies of N20, N30 and P40 throughout the expiratory loading period, except for a significant lengthening in P1 latency compared with unloaded breathing. However, inspiratory loading induced a significant increase in peak latency of N20, N30 and P40 components. We suggest that projections of inspiratory afferents from the diaphragm and the intercostal muscles, activated by inspiratory loading, could be responsible for the lengthened latency of median nerve SEP components. Thus, respiratory afferents very likely interact with pathways of the somatosensory system. PMID- 10462119 TI - Electrophysiological actions of N-[1-[4-(4-fluorophenoxy)butyl]-4-piperidinyl]-N methyl-2-benzothiazola mine (R56865) on CA1 neurons of the rat hippocampal slice during hypoxia. AB - The electrophysiological effects of N-[1-[4-(4-fluorophenoxy)butyl]-4 piperidinyl]-N-methyl-2-benzothiazo lamine (R56865), a drug which protects heart cells from ischemia-induced arrhythmias, was studied on intracellularly-recorded CA1 neurons of the rat hippocampal slice under normal or hypoxic conditions. On normoxic cells R56865 (1 microM) reduced firing accommodation without changing passive membrane properties, spike characteristics or synaptic transmission. On hypoxic cells R56865 selectively reduced the amplitude of hypoxia-induced membrane depolarization and partly counteracted the depression of synaptic transmission evoked by Schaffers collateral stimulation. Despite its influence on repetitive firing properties, R56865 might be useful to limit the extent of cellular depolarizing responses to hypoxia. PMID- 10462120 TI - Transforming growth factor beta2 is able to modify mRNA levels and release of luteinizing hormone-releasing hormone in a immortalized hypothalamic cell line (GT1-1). AB - On the basis of our previous observations which indicated that transforming growth factor beta1 (TGFbeta1) affects the gene expression and the release of luteinizing hormone-releasing hormone (LHRH) in GT1-1 cells, we have presently evaluated whether also TGFbeta2 might be effective on these parameters. The data here reported show that also TGFbeta2 is able to affect LHRH dynamics, and that this action presents a different kinetics than that reported by TGFbeta1. In particular TGFbeta2 is able to facilitate LHRH release and to decrease the mRNA levels of this decapeptide. The present data have also shown that, GT1-1 cells express the messengers for the two most important receptors of the TGFbeta family, namely TGFbetaRI and TGFbetaRII and consequently represent a target for the action of the different isoforms of TGFbeta. Since the two isoforms of TGFbeta are produced and released from astrocytes, the present data add new support to the hypothesis that astrocytes participate in the control of LHRH secretion in a paracrine fashion. PMID- 10462121 TI - Enhanced inhibition of free radical-induced deoxyribose breakdown by Alzheimer brain homogenates. AB - The ability of homogenates from Alzheimer and control brains to inhibit formation of thiobarbituric acid reactive products (TBAR) induced by free radicals was compared. The assay for TBAR was modified by adding 1% sodium dodecyl sulfate (SDS) to prevent chromogen adsorption by biological matrices, and by extending the incubation time. The inhibitory activities required smaller equivalents of Alzheimer brain homogenates than control homogenates. Similar inhibitory activities were seen in homogenates from amygdala, temporal cortex and cerebellum. The inhibitory activities were similar in brain homogenates from individuals with different apolipoprotein E status. These results indicate that Alzheimer brain tissue has either increased content of free radical scavengers or is more sensitive to free radical attack than control brains. PMID- 10462122 TI - Scopolamine-sensitive and resistant components of increase in cerebral cortical blood flow elicited by periaqueductal gray matter of rats. AB - The present study attempted to evaluate the effects of inhibition of cortical muscarinic receptors on the increase in cortical blood flow (laser-Doppler flowmetry) elicited by chemical stimulation of the periaqueductal gray matter in 28 anesthetized rats with cervical cordotomy. A new device was introduced which allowed focal cortical superfusion with scopolamine, an antagonist for muscarinic receptors, without disturbing the temperature of an exposed cortical area under study. We found that although the flow increase was attenuated by scopolamine (31.6 micromol/l to 1 mmol/l) so applied, about one third of it was resistant to the treatment. Cortical muscarinic and non-muscarinic receptor mechanisms may thus subserve the mediation of the flow increase. PMID- 10462123 TI - Adenovirus-mediated gene transfer of Bcl-xL prevents cell death in primary neuronal culture of the rat. AB - Bcl-xL is a Bcl-2-related gene that regulates programmed cell death in a bcl-2 independent fashion. It is expressed in tissues containing long-surviving postmitotic cells, such as neurons in adult brains. To investigate the possibility of gene therapy for transferring this anti-apoptotic gene into the neuron for the treatment of vascular occlusive or neurodegenerative diseases, we examined the effect of a replication-deficient recombinant adenovirus vector coding human Bcl-xL gene on the augmentation of the survival of primarily cultured rat neuronal cells in vitro. Immunoblot analysis revealed that primarily cultured neuronal cells were successfully infected and transferred with this gene by recombinant adenovirus vector with high transduction efficiency. Bcl-xL gene transfer to the primarily-cultured neurons could prevent these cells from cell death. PMID- 10462124 TI - Activation of sensory nerves participates in stress-induced histamine release from mast cells in rats. AB - To elucidate the mechanism by which stress induces rapid histamine release from mast cells, Wistar rats, pretreated as neonates with capsaicin, were subjected to immobilization stress for 2 h, and histamine release was measured in paws of anesthetized rats by using in vivo microdialysis after activation of sensory nerves by electrical or chemical stimulation. Immobilization stress studies indicated that in control rats stress induced a 2.7-fold increase in the level of plasma histamine compared to that in freely moving rats. Whereas pretreatment with capsaicin significantly decreased stress-induced elevation of plasma histamine. Microdialysis studies showed that electrical stimulation of the sciatic nerve resulted in a 4-fold increase of histamine release in rat paws. However, this increase was significantly inhibited in rats pretreated with capsaicin. Furthermore, injection of capsaicin into rat paw significantly increased histamine release in a dose-dependent manner. These results suggest that activation of sensory nerves participates in stress-induced histamine release from mast cells. PMID- 10462125 TI - Cortical focusing is an alternative explanation for improved sensory acuity on an amputation stump. AB - The ability to localize a sensory stimulus on the body surface (locognosia) has been investigated in normal subjects in a quest to find an explanation for the reported findings of improved sensory acuity on an amputation stump. We have shown that when attending to a smaller area of skin, during the testing procedure, locognosia improves (P < 0.001) by a similar degree to that seen in amputees. Such selective attention is likely to occur in upper limb amputees as they have a reduced area of skin on which to focus during sensory testing. This represents a further explanation for improved sensory acuity on an amputation stump without implicating plasticity of connections within the somatosensory cortex. PMID- 10462126 TI - Acute and transient increase of lipocalin-type prostaglandin D synthase (beta trace) level in cerebrospinal fluid of patients with aneurysmal subarachnoid hemorrhage. AB - We measured the concentration of lipocalin-type prostaglandin D synthase (PGDS) in cerebrospinal fluid (CSF) and serum in patients 1, 3, 5, 7, 9, 11, 14 and 17 days after subarachnoid hemorrhage (SAH) due to ruptured cerebral aneurysms. The PGDS level in lumbar CSF increased about two-fold at day 3 (20.85 +/- 2.71 microg/ml, mean +/- SE) and at day 5 (25.24 +/- 3.76), as compared with the level at day 1 (11.25 +/- 1.07). The CSF level gradually decreased and returned to the day 1 level at day 17. The serum PGDS level was much lower than the CSF level (0.39 +/- 0.06 at day 1) and almost unchanged until day 17. The neuron-specific enolase level in CSF, as an index of brain damage, was maximum at day 1 (29.83 +/ 7.32 ng/ml) and decreased at day 3 and at day 5 (18.28 +/- 2.65 and 11.95 +/- 1.82, respectively). These results suggest that the transient and delayed increase in the PGDS level in CSF is due to its induction of PGDS in the arachnoid membrane after SAH. PMID- 10462127 TI - A review of central 5-HT receptors and their function. AB - It is now nearly 5 years since the last of the currently recognised 5-HT receptors was identified in terms of its cDNA sequence. Over this period, much effort has been directed towards understanding the function attributable to individual 5-HT receptors in the brain. This has been helped, in part, by the synthesis of a number of compounds that selectively interact with individual 5-HT receptor subtypes--although some 5-HT receptors still lack any selective ligands (e.g. 5-ht1E, 5-ht5A and 5-ht5B receptors). The present review provides background information for each 5-HT receptor subtype and subsequently reviews in more detail the functional responses attributed to each receptor in the brain. Clearly this latter area has moved forward in recent years and this progression is likely to continue given the level of interest associated with the actions of 5-HT. This interest is stimulated by the belief that pharmacological manipulation of the central 5-HT system will have therapeutic potential. In support of which, a number of 5-HT receptor ligands are currently utilised, or are in clinical development, to reduce the symptoms of CNS dysfunction. PMID- 10462128 TI - Effects of venlafaxine on extracellular concentrations of 5-HT and noradrenaline in the rat frontal cortex: augmentation via 5-HT1A receptor antagonism. AB - Venlafaxine is a novel serotonin/noradrenaline reuptake inhibitor (SNRI) which has been shown clinically to be an effective antidepressant (AD) with a faster onset of action than serotonin specific reuptake inhibitors (SSRI). Preclinically, venlafaxine has been shown to potently inhibit dorsal raphe neuronal (DRN) firing through a 5-HT1A mediated mechanism, in a similar manner to SSRIs. Here we demonstrate the acute neurochemical effects of venlafaxine on extracellular concentrations of 5-HT and noradrenaline (NA) from the rat frontal cortex using in vivo microdialysis. Administration of venlafaxine (3-50 mg/kg s.c.) resulted in a significant dose-dependent increase in extracellular NA, but produced no significant increase in 5-HT concentrations. Combination treatment with the selective 5-HT1A antagonist WAY100635 produced a dose-dependent augmentation of venlafaxine-induced (3-30 mg/kg s.c) extracellular 5-HT concentrations, but had no further effect on NA above that produced by venlafaxine alone. WAY100635, at doses as low as 0.03 mg/kg s.c., maintained this potentiation effect. The beta-adrenergic/5-HT1A receptor antagonist (+/-)pindolol and the selective 5-HT1B/D antagonist GR127935 produced no significant augmentation of venlafaxine-induced changes in either 5-HT or NA. Using the alpha1 and alpha2-adrenoceptor antagonists, prazosin and idazoxane, we also demonstrate the role of the alpha-adrenoceptors in the augmentation of venlafaxine-induced changes. The possible mechanisms underlying venlafaxines improved clinical AD action and the potential for further enhancement of this SNRIs clinical effects are discussed. PMID- 10462129 TI - WAY 100635, a 5-HT1A receptor antagonist, prevents the impairment of spatial learning caused by blockade of hippocampal NMDA receptors. AB - We studied the effect of WAY 100635, a 5-HT1A receptor antagonist, on the impairment of spatial learning caused by intrahippocampal injection of 3-((R)-2 carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP), a competitive NMDA receptor antagonist, in a two-platform spatial discrimination task. CPP, 3 and 10 ng/microl, administered bilaterally into the CA1 region of the dorsal hippocampus 10 min before each training session, dose-dependently reduced choice accuracy in the two-platform spatial discrimination task with little or no effect on choice latency and errors of omission. A volume of 10 ng/microl intrahippocampal CPP did not affect choice accuracy or latency of a non-spatial visual discrimination task. Subcutaneous doses of 0.3 and 1 mg/kg WAY 100635 did not modify the choice accuracy, but prevented the impairment caused by 10 ng/microl intrahippocampal CPP. A dose of 20 ng/microl WAY 100635 into the dorsal hippocampus prevented the deficit caused by 10 ng/microl CPP administered in the same region. The results suggest that blockade of 5-HT1A receptors can compensate the loss of NMDA mediated excitatory input to pyramidal cells in the hippocampus. These findings may have clinical relevance for the symptomatic treatment of memory disorders associated with reduced glutamate transmission mediated by NMDA receptors. PMID- 10462130 TI - The antipsychotic drug risperidone interacts with auto- and hetero-receptors regulating serotonin output in the rat frontal cortex. AB - We have previously shown that the antipsychotic drug risperidone enhances serotonin (5-HT) output in the rat frontal cortex (FC), but the precise underlying mechanism has not been revealed. Consequently, the present study using in vivo microdialysis was undertaken to (i) characterize the effects of alpha2D, 5-HT1B and 5-HT1D receptor stimulation or blockade on 5-HT efflux in the FC given the purported regulatory role of these sites on 5-HT release, and (ii) to investigate the ability of risperidone to interfere with these receptors in order to examine their putative role in the facilitatory action or risperidone on cortical 5-HT output. Cortical perfusion with risperidone or the alpha2A/D, 5 HT1B and 5-HT1B/1D receptor antagonists idazoxan, isamoltane or GR 127,935, respectively, dose-dependently increased 5-HT efflux in the FC. Conversely, agonists at these receptors, i.e. clonidine, CP 93,129 or CP 135,807, respectively, decreased extracellular 5-HT concentrations. The agonist-induced decreases in 5-HT efflux were antagonized by coadministration of respective receptor antagonists. Risperidone attenuated the decrease in cortical 5-HT efflux elicited by clonidine or CP 135,807 but failed to affect the decrease elicited by CP 93,129. The present in vivo biochemical data indicate that the output of 5-HT in the FC is negatively regulated via alpha2D, 5-HT1B and tentatively also via 5 HT1D receptors located in the nerve terminal area. Moreover, the results indicate that risperidone acts as an antagonist at alpha2D and possibly 5-HT1D receptors in vivo, two properties which most likely contribute to its stimulatory effect on cortical 5-HT efflux. The facilitatory effect of risperidone on cortical serotonergic neurotransmission may be of significance for its therapeutic effect in schizophrenia, particularly when associated with affective symptomatology and/or intense anxiety. The effect may also contribute to alleviate signs of cortical dysfunction such as impaired cognition. PMID- 10462131 TI - In vivo receptor occupancy of NRA0045, a putative atypical antipsychotic, in rats. AB - We have previously reported that (R)-(+)-2-amino-4-(4-fluorophenyl)-5-[1-[4-(4 fluorophenyl)-4-oxobutyl]+ ++pyrrolidin-3-yl]thiazole (NRA0045) is a novel antipsychotic agent with affinities for dopamine D4, 5-hydroxytryptamine 2A (5 HT2A) and alpha1 receptors. In the present study, in vivo receptor occupancy of 5 HT2A, alpha1, dopamine D2 and D3 receptors by NRA0045 was assessed, based on in vivo and ex vivo receptor binding, and findings were compared to reference antipsychotic drugs (haloperidol, risperidone, clozapine). Intraperitoneal administration of haloperidol highly occupied the dopamine D2 receptor in the striatum and nucleus accumbens, and alpha1 adrenoceptors in the frontal cortex. Occupation of the 5-HT2A receptor in the frontal cortex and the dopamine D3 receptor in the nucleus accumbens and islands of Cajella was moderate. By contrast, atypical antipsychotics such as risperidone and clozapine dose dependently occupied the 5-HT2A receptor in the frontal cortex, with moderate to negligible occupancy of the D2 receptor in the striatum and the nucleus accumbens. Clozapine and risperidone also occupied the alpha1 adrenoceptor in the frontal cortex, and clozapine did not occupy the dopamine D3 receptor. As seen with other atypical antipsychotics, intraperitoneal administration of NRA0045 dose-dependently occupied the 5-HT2A receptor and the alpha1 adrenoceptor in the frontal cortex, while it was without effect on dopamine D2 and D3 receptors in the striatum, nucleus accumbens and islands of Cajella. Thus, the strong occupancy of 5-HT2A and alpha1 receptors is involved in the pharmacological action of NRA0045. PMID- 10462132 TI - SB 242084, a selective serotonin2C receptor antagonist, increases dopaminergic transmission in the mesolimbic system. AB - Electrophysiological techniques and in vivo microdialysis were used to investigate the effect of SB 242084, a potent and selective 5-HT2C receptor antagonist in the control of nigro-striatal and mesolimbic dopaminergic function. Thus, extracellular single unit recordings were performed from neurochemically identified dopamine (DA) neurons in the substantia nigra, pars compacta (SNc) and the ventral tegmental area (VTA), as well as monitoring of striatal and accumbal basal DA release in anesthetized rats following the administration of SB 242084 and RO 60-0175. Administration of SB 242084 (160-640 microg/kg, i.v.) caused a dose-dependent increase in the basal firing rate of VTA DA neurons, reaching its maximum (27.8+/-6%, above baseline) after 640 microg/kg. Moreover, bursting activity was significantly enhanced by SB 242084 in the VTA. On the other hand, SB 242084 (160-640 microg/kg, i.v.) did not cause any significant change in the basal firing rate and bursting activity of DA neurons in the SNc. Injection of the 5-HT2C receptor agonist RO 60-0175 (80-320% microg/kg, i.v.) dose-dependently decreased the basal firing of DA neurons in the VTA but not in the SNc. RO 60 0175 exerted its maximal inhibitory effect (53.9+/-15.1%, below baseline) in the VTA at the dose of 320 microg/kg. Basal DA release (34.8+/-9%, above baseline) and dihydroxyphenylacetic acid (DOPAC) efflux (19.7+/-7%, above baseline) were significantly enhanced in the nucleus accumbens following the intraperitoneal administration of 10 mg/kg SB 242084. Intraperitoneal injection of 5 mg/kg SB 242084 significantly increased DA release (16.4+/-6%, above baseline) in the nucleus accumbens, but did not affect DOPAC efflux. In the striatum, SB 242084 (5 and 10 mg/kg, i.p.) only slightly increased DA release above baseline (3.5+/-4 and 11.2+/-6%, respectively), without affecting DOPAC efflux in this area. However, the effect of SB 242084 in the striatum was rendered more evident by the fact that injection of the vehicle used to dissolve the drug in a group of control rats, significantly reduced basal DA output by 19.6+/-7%. Stimulation of 5-HT2C receptors by RO 60-0175 (1 mg/kg, i.p.) significantly decreased DA release in the nucleus accumbens by 26.1+/-4% (below baseline) 60 min after injection. On the other hand, RO 60-0175 (1 mg/kg, i.p.) did not cause any significant change of DA release in the striatum. However, DOPAC efflux was reduced by RO 60-0175 (1 mg/kg, i.p.) both in the striatum and the nucleus accumbens. Taken together, these data indicate that the central 5-HT system exerts a tonic and phasic inhibitory control on mesolimbic DA neuron activity and that 5-HT2C receptor subtypes are involved in this effect. Moreover, these findings might open new possibilities for the employment of 5-HT2C receptor antagonists in the treatment of neuropsychiatric disorders related to a hypofunction of central DA neurons. PMID- 10462133 TI - Social behavior of juvenile rats after in utero exposure to morphine: dose-time effect relationship. AB - In the present study, the effects of morphine exposure in utero on social behavior in juvenile male rats was investigated. Pinning, a measure for play behavior, and social grooming of the offspring were measured at postnatal day 21. The subjects were offspring of Wistar rat dams given sc. injections of 1 or 10 mg/kg body weight morphine HCl daily from gestational days 8 (GD8)-GD 21 and control dams injected daily with saline. Pinning and social grooming of the morphine-treated offspring were significantly elevated compared to saline controls. The doses of morphine used neither affected the gestation of pregnant mother rats nor sensorimotor development of the juvenile rats. Prenatal exposure to morphine of 10 mg/kg daily increased both pinning and social grooming, prenatal exposure to a lower dose of 1 mg/kg increased pinning behavior but not social grooming in the offspring. To study the importance of the gestational period, offspring of dams given 10 mg/kg body weight morphine HCl from GD8-GD15 and saline from GD16-parturition or morphine from GD16-parturition and saline from GD8-GD15 was tested. Pinning was only increased when morphine exposure occurred during the third week of gestation, social grooming was increased when morphine exposure had been in the second week of gestation. Subcutaneous administration of 1 mg/kg naltrexone 1 h before the test significantly decreased play behavior in control rats, but not in animals prenatally exposed to morphine. From these experiments we conclude that the long term effect of in utero exposure to morphine on play behavior is established by affecting the endogenous opioid system. PMID- 10462134 TI - Quinolinic acid formation in immune-activated mice: studies with (m-nitrobenzoyl) alanine (mNBA) and 3,4-dimethoxy-[-N-4-(-3-nitrophenyl)thiazol-2yl]-benzenesul fonamide (Ro 61-8048), two potent and selective inhibitors of kynurenine hydroxylase. AB - The role of kynurenine hydroxylase activity in the neo-formation of the excitotoxin quinolinic acid (QUIN) has been studied in mice by using (m nitrobenzoyl)-alanine (mNBA) and 3,4-dimethoxy-[-N-4-(-3-nitrophenyl)thiazol-2yl] benzenesulf onamide (Ro 61-8048), two potent and selective inhibitors of this enzyme. Immune-stimulation with pokeweed mitogen (PWM, 200 microg i.v., 12 h) induced a robust increase in kynurenine (KYN) and its metabolites kynurenic acid (KYNA) and QUIN in blood and brain. When incubated in a medium containing KYN but not tryptophan, spleen, lung and liver (but not brain) slices accumulated a measurable amount of QUIN in the supernatant. Slices obtained from PWM treated animals had a ten-fold increase in QUIN accumulation in spleen, no changes in lung and a 40% decrease in liver, suggesting that the spleen contributes to the increased QUIN levels found in the blood and brain of immune-stimulated mice. Large doses of kynurenine hydroxylase inhibitors increased KYN and KYNA, but unexpectedly did not decrease QUIN content in control blood and brain. When tested in organ slices obtained from either controls or immune-stimulated animals, mNBA (1-1000 microM) and Ro 61-8048 (0.1-100 microM) strongly reduced QUIN neo-formation, suggesting that, in vitro, kynurenine hydroxylase activity is required for QUIN neosynthesis. Indeed, after repeated doses of mNBA or Ro 61 8048, QUIN content in blood and brain of immune-stimulated animals significantly decreased. Our results suggest that, under basal conditions, sufficient QUIN synthesis may occur through kynurenine hydroxylase-independent pathways. In immune-stimulated animals, however, kynurenine hydroxylase inhibitors significantly reduce blood and brain accumulation of QUIN. PMID- 10462135 TI - Blockade of NMDA channels in acutely isolated rat hippocampal neurons by the Na+/Ca2+ exchange inhibitor KB-R7943. AB - Neurons acutely isolated from the CA1 region of rat hippocampal slices using the 'vibrodissociation' method were voltage-clamped in the whole-cell configuration. The currents through NMDA channels were elicited by application of 100 microM aspartate (ASP) in a Mg2+-free solution in the presence of 3 microM glycine. The compound KB-R7943, (2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl]isothiourea methanesulfonate) known as a Na+/Ca2+ exchange inhibitor was able to block effectively the NMDA channels. At a holding potential of -100 mV, the measurement of the concentration dependence of the stationary current blockade revealed the existence of two populations of NMDA channels differing by a high (IC50 = 0.8 microM) and low (IC50 = 11 microM) affinity for KB-R7943. The Hill coefficients indicated that one blocking molecule can bind to NMDA channels which have a high affinity for KB-R7943 and at least two blocking molecules can bind to the NMDA channels which have a low affinity for KB-R7943. When applied externally, KB R7943 can bind to the low-affinity NMDA channels irrespective of whether or not these channels are activated by the agonist. The KB-R7943-induced blockade of the NMDA channel was partly voltage-dependent. Within the framework of the Woodhull model, the apparent value of delta calculated for the voltage-dependent binding of KB-R7943 was in the range of 0.26-0.41. The blocking action of KB-R7943 on NMDA channels did not depend either on ASP or glycine concentrations which indicated that the binding sites for KB-R7943 and those for the agonist and the coagonist did not overlap. PMID- 10462136 TI - Beta-amyloid-induced apoptosis of cerebellar granule cells and cortical neurons: exacerbation by selective inhibition of group I metabotropic glutamate receptors. AB - Administration of beta-amyloid fragment 25-35 (Abeta25-35) to cultured rat cerebellar granule cells (CGC) or cortical neurons caused cell death that was characterized by morphological and nuclear changes consistent with apoptosis. Inhibition of NMDA receptors produced a mild exacerbation of Abeta25-35 toxicity in cortical neurons; a similar effect was induced by AMPA/kainate receptor inhibition in CGC. Selective activation of group I metabotropic glutamate receptors (mGluR) by dihyroxyphenylglycine (DHPG) had no effect on Abeta25-35 induced apoptosis in either cell type, and was unaffected by blockade of ionotropic glutamate receptors. In contrast, selective inhibition of group I mGluR by (RS)-1-aminoindan-1,5-dicarboxylic acid (AIDA) exacerbated Abeta toxicity in cortical neurons, whereas this treatment was without effect on CGC. However, AIDA significantly increased Abeta-induced apoptosis in CGC in the presence of either NMDA or AMPA/kainate receptor inhibition; blockade of both ionotropic glutamate receptor classes further increased the exacerbation of apoptosis following treatment with AIDA. These findings suggest that Abeta25-35 induced neuronal injury leads to activation of group I mGluR, which attenuates the resulting apoptosis. PMID- 10462138 TI - Absence of toxicity associated with adenoviral-mediated transfer of the beta galactosidase reporter gene to neonatal rat islets in vitro. AB - Transfer of genes with potential therapeutic utility to the pancreatic islets of Langerhans may enhance graft survival after islet transplantation. The aim of this study was to determine the optimal conditions for adenoviral-mediated gene transfer to the islets of Langerhans in the absence of vector-induced toxicity. Neonatal rat islets were transduced in groups of 25 with an adenoviral vector encoding beta-galactosidase (AdbetaGal) at doses of MOI 0, 10, 100 and 1000 pfu per islet cell. All experiments were performed in triplicate. Efficiency of gene transfer was determined by gross inspection and estimation of the percentage of beta-galactosidase positive cells after islet dispersion at 1, 4, 7 and 10 days post-transduction. Islet toxicity was assessed by measuring accumulated insulin levels at each time-point and by assessing static incubation insulin release at 3 and 10 days. Efficient dose-dependent gene transfer to the islets was documented at 1, 4, 7 and 10 days post-transduction. Transgene expression was relatively stable for the duration of the experiment. Insulin accumulation did not differ between transduced and non-transduced islets at each timepoint. Likewise, the insulin secretory response to glucose, obtained by dividing the insulin response to high glucose incubation by the insulin response to low glucose incubation was similar in transduced and non-transduced islets at 3 and 10 days at all doses studied. In summary, adenoviral-mediated transduction of islets results in dose dependent efficient gene transfer with relatively stable transgene expression in the absence of toxicity. This technology may be useful in the study of islet biology and also in the future in gene therapy approaches to the treatment of diabetes mellitus. PMID- 10462137 TI - Constrictor responses of resistance arterioles during diabetes mellitus. AB - While diabetes mellitus appears to alter nitric oxide synthase-dependent vasodilatation, the effect of diabetes on constrictor responses of resistance arterioles is not clear. Our goal was to examine the effect of diabetes on constrictor responses of cheek pouch arterioles. In vivo diameter of arterioles ( approximately 50 microm) was measured in response to norepinephrine, the thromboxane analogue (U-46619) and endothelin-1 in nondiabetic and diabetic hamsters (4-6 weeks post streptozotocin). Norepinephrine (1.0 and 10 nM) and U 46619 (0.1 and 1.0 nM) produced similar dose-related vasoconstriction in nondiabetic and diabetic hamsters (P > 0.05). In contrast, vasoconstriction to endothelin-1 (0.1 and 1.0 pM) was greater in diabetic than nondiabetic hamsters (P < 0.05). Next, we examined the role of nitric oxide in basal vascular tone and whether enhanced vasoconstriction in diabetic hamsters to endothelin-1 might be related to an alteration in the modulatory role of nitric oxide. N(G)-monomethyl L-arginine (L-NMMA) (1.0, 10 and 100 microM) produced dose-related vasoconstriction in nondiabetic, but not diabetic hamsters. Further, L-NMMA did not alter vasoconstriction in response to endothelin-1 in nondiabetic and diabetic hamsters. These findings suggest that diabetes alters constriction of cheek pouch resistance arterioles to endothelin-1 which appears to be independent of the synthesis/release of nitric oxide. In addition, based upon findings using L-NMMA, it appears that there is a reduced influence of nitric oxide on basal diameter of resistance arterioles during diabetes. PMID- 10462139 TI - Long-term follow-up of insulin-dependent diabetes mellitus patients with recurrent episodes of severe hypoglycaemia. AB - We conducted a long-term clinical follow-up of 20 insulin-treated diabetic patients with the problem of repeated severe hypoglycaemic events, identified during 1982-1984. Furthermore, we investigated the reproducibility of a standardized insulin-infusion test among eight of these patients and 10 insulin dependent diabetes mellitus patients without the problem of severe hypoglycaemia. At start of the follow-up, the patients were characterized, which revealed that proposed risk factors for severe hypoglycaemia were very common and that at least one predisposing factor was present in all patients. No fatal episode of hypoglycaemia occurred in this group of patients during the follow-up. Emergency visits due to hypoglycaemia decreased during the follow-up from 1.05 year(-1) in 1982-1984 to 0.42 year(-1) in 1992-1994 (P=0.024). No association to HbAlc was seen (P=0.11). A correlation with the change to multiple insulin regimen was seen with a incidence of 1.27 year(-1) before the change to 0.47 year(-1) after (P=0.003). The glucose response to an insulin-infusion test appeared reproducible among the patients with recurrent attacks of severe hypoglycaemia. PMID- 10462140 TI - Evaluation of a real-time blood glucose monitor in children with diabetic ketoacidosis. AB - Use of a real-time bedside glucose monitor was analyzed during the course of management of diabetic ketoacidosis (DKA) in children. Simultaneous determinations of blood glucose were obtained, using three methods: bedside glucose meter (One Touch II), laboratory glucose analyzer (YSI 2300 STAT), and a real-time bedside glucose monitor (VIA 1-01G Blood Chemistry monitor). Study patients included seventeen patients < 18 years of age admitted to a Pediatric Intensive Care Unit, with blood samples obtained during treatment of DKA by continuous insulin infusion. Four patients did not complete the study. Three experienced temporary technical problems with the monitor, and four required repeat IV placement. Duration of monitor use ranged between 6 and 47 h (mean 24 +/- 4 h). Blood glucose values ranged between 2.6 and 22.5 mmol/l. Overall correlation of blood glucose values were as follows: 0.965, 0.965, 0.973, VIA 1 01G vs. One Touch II, VIA 1-01G vs. YSI 2300 STAT, and One Touch II vs. YSI 2300 STAT, respectively (all P-values < 0.0001). This real-time bedside glucose monitor is accurate at glucose values < 13.8 mmol/l, and reliable for rapid, repetitive analyses. Results indicate that blood glucose values obtained using this real-time monitor are comparable to those using standard methods of measurement, and that this device is clinically applicable for use in management of children with DKA. PMID- 10462141 TI - Renal complications in patients with diabetes mellitus associated with an A to G mutation of mitochondrial DNA at the 3243 position of leucine tRNA. AB - The substitution of guanine for adenine at position 3243 of the leucine tRNA gene of mitochondrial DNA was originally described in association with MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes). Diabetes mellitus associated with the mutation (mitochondrial diabetes) is a different phenotype from MELAS. We identified 11 patients with the mutation among 385 Japanese diabetic patients: two had MELAS and nine had mitochondrial diabetes. We present data on a male patient with mitochondrial diabetes who developed the nephrotic syndrome at the age of 23. Light microscopy revealed mesangial expansion, PAS-positive deposits and segmental sclerosis in the glomeruli. Scattered mesangial electron-dense deposits and thickening of the basement membrane were found on electron microscopy, suggesting that diabetic glomerulosclerosis accompanied by focal glomerulosclerosis (FGS). Mitochondrial diabetes may pre-dispose patients to renal complications, including forms of glomerulonephritis, such as FGS. PMID- 10462142 TI - Microalbuminuria and retinopathy in a diabetic population of Cameroon. AB - Microalbuminuria and retinopathy was studied in a non-proteinuric diabetic population of Cameroon. Patients were enrolled on a consecutive basis in two referral hospitals in Yaounde. Retinopathy was evaluated by direct ophthalmoscopy and biomicroscopy, and controlled by mydriatic fundus photography. Detection of microalbuminuria was carried out on an overnight urine sample using Micral II test (Boehringer Mannheim). Anthropometric and blood pressure measurements were done using validated methods. In 64 non-proteinuric diabetic patients (9 IDDM and 55 NIDDM) aged 19-70 years with known duration of diabetes of 1-23 years, the prevalence of retinopathy was 37.5%. Microalbuminuria was detected in 53.1% of patients. Microalbuminuria correlated with duration of diabetes, and blood pressure, retinopathy was positively correlated with age, and blood pressure. Retinopathy was not significantly associated with the known duration of diabetes. Retinopathy was found to be independently associated with microalbuminuria (P < 0.001) and microalbuminuria appeared to be a sensitive marker of retinopathy. The prevalence of retinopathy and microalbuminuria in this population was high. Microalbuminuria and non-proliferative retinopathy are independently associated, and are both associated with increased blood pressure levels in the study population. As shown in previous studies microalbuminuria may also be a sensitive marker of early diabetic retinopathy in African diabetic patients. PMID- 10462143 TI - Cilnidipine, the N- and L-type calcium channel antagonist, reduced on 24-h urinary catecholamines and C-peptide in hypertensive non-insulin-dependent diabetes mellitus. AB - To evaluate the effects of cilnidipine (CNP), L- and N-type calcium channel blocker and nilvadipine (NVP) on 24-h urinary epinephrine (U-EP), norepinephrine (U-NE), dopamine (U-DA) and C-peptide (U-CPR) in patients associated with hypertension and non-insulin-dependent diabetes mellitus (HT-NIDDM), a randomized crossover study was performed with 35 HT-NIDDM patients. The patients were given CNP (10 mg/day) and NVP (8 mg/day), separately, for 4 weeks each. After CNP treatment, U-NE, U-DA and U-CPR levels were significantly reduced compared with pre-treatment levels: 160.4 +/- 12.7 to 111.7 +/- 8.9 microg/day (mean +/- S.E., P < 0.005); 934.8 +/- 163.4 to 590.3 +/- 33.4 microg/day (P < 0.05); 86.7 +/- 9.9 to 57.6 +/- 7.4 microg/day (P < 0.05), respectively. Although no significant differences were observed in U-EP, U-NE, U-DA and U-CPR levels by NVP treatment, U-NE, U-DA and U-CPR levels after CNP treatment were significantly lower than those after NVP treatment: 111.7 +/- 8.9 versus 155.0 +/- 13.7 microg/day (P < 0.02); 590.3 + 33.4 versus 822.2 +/- 104.3 microg/day (P < 0.05); 57.6 +/- 7.4 versus 80.6 +/- 8.1 microg/day (P < 0.05), respectively. In conclusion, it was demonstrated that CNP treatment significantly reduced U-NE, U-DA and U-CPR excretion compared with NVP treatment in HT-NIDDM patients. PMID- 10462144 TI - Impacts of urbanisation on the lifestyle and on the prevalence of diabetes in native Asian Indian population. AB - Recent studies from the Asian subcontinent show an increasing prevalence of diabetes. This increase has been attributed to factors related to lifestyle changes related to modernisation. A periurban rural population resembling the rural in their occupation, but with access to certain urban facilities was chosen for this study. The aim of the study was to assess the impact of modernisation on the rising prevalence of diabetes in the native Indians. A total of 1637 adults aged 20 years and above (749 men and 888 women) were tested for diabetes and impaired glucose tolerance (IGT) by 2 h post-glucose challenge. Demographic, anthropometric, dietary and occupational details, were recorded. Dietary habits were similar in all categories of socio-economic strata. In the present study group, the age standardised prevalence of Type 2 diabetes was 5.9%, which was intermediate to that in the urban (11.6%) and rural (2.4%) populations. The prevalence data of the latter two population were available from previous surveys. Prevalence of impaired glucose tolerance (IGT) was high (6.9%) and similar in all three population samples. In the periurban population, a large percentage of subjects were doing only routine household work and had a sedentary life-style. After correcting for the age and BMI, sedentary work and occupation had a significant association with diabetes, suggesting that sedentary lifestyle may be an important determinant for the higher prevalence of diabetes in an urbanising population. PMID- 10462145 TI - Mild head injury: a misnomer. AB - Despite controversy surrounding the concept of mild head injury (MHI), it is becoming evident that even a head trauma termed 'mild' may result in significant behavioural sequelae. The present study was an attempt at documenting structural cerebral damage, by way of computerized tomography, in a group of patients having suffered a MHI as defined by the Glasgow Coma Scale (GCS) score. A 1-year retrospective chart review identified 80 MHI patients who presented to the Emergency department of a lead hospital for trauma. Sixty-six per cent of these MHI patients were scanned. Evidence of intracranial abnormalities was obtained in 31% of the overall sample. Patients with a lower GCS score had a higher percentage of abnormal scans than those with a GCS score of either 14 or 15. The present findings suggest that a MHI can be associated with significant morbidity, and that a MHI group does not constitute a homogeneous pool of patients. PMID- 10462146 TI - Exploratory investigation into mild brain injury and discriminant analysis with high frequency bands (32-64 Hz). AB - QEEG variables (five activation, two relationship variables, 19 locations and five bands up to 64 Hertz) were collected under eyes closed condition (under both 32 and 64 Hertz conditions) on 91 subjects, consisting of 32 mild brain-injured subjects (no loss of consciousness greater than 20 minutes) and 52 normals over the age of 14. An additional seven subjects who were unconscious greater than 20 minutes were available for analysis. Previous discriminant function analysis developed by Thatcher et al. was employed on the eyes closed 32 Hertz condition to ascertain its robustness for time periods greater than 1 year and for significant periods of unconsciousness. A separate discriminant for subjects was developed, employing only frontal high frequency coherence figures. The Thatcher discriminant could reliably (79%) identify all subjects up to 43 years post accident. The high frequency discriminant effectively identified 87% of the brain injured across all time periods (without significant loss of consciousness) and 100% of subjects within 1 year of accident. The combination of the discriminants resulted in a 100% accuracy rate for the 39 brain injured subjects for which discriminate values were available. PMID- 10462147 TI - Cognitive and psychosocial outcome following moderate or severe traumatic brain injury. AB - The outcome of 96 consecutive adult patients with moderate to severe head injury was sequentially measured at 6, 12 and 24 months post-injury. In addition to global outcome using the Glasgow Outcome Scale (GOS) and a battery of neuropsychological tests of cognitive function, the Head Injury Symptom Checklist (HISC) and Relative's Questionnaire (RQ) were used. Although poorer GOS scores and severe cognitive impairments were typically associated with greater severity of initial injury, relatives reported similar functional problems irrespective of injury severity. This illustrates the legacy of moderate head injury in influencing many aspects of everyday life, supporting the argument that the needs of this group should not be overlooked. PMID- 10462148 TI - Measuring recovery from post traumatic amnesia. AB - Three groups of participants were assessed. Each participant was tested on 20 occasions. The groups comprised people (i) in post traumatic amnesia (PTA) following severe head injury (n=9), (it) with severe head injury but not in PTA (n=10), and (iii) with no history of head injury or other neurological condition (n=13). Subjects were given several tests of memory, attention and learning in order to determine which tests were good at (a) distinguishing people in PTA from those not in PTA, and (b) monitoring recovery over time. The results indicate that people in PTA have a wide range of deficits and their cognitive recovery is a gradual process rather than an all-or-none phenomenon. In terms of measurement, the study suggests that a good test of PTA should include orientation questions, together with a reaction time measure, a visual recognition test and a speed of information processing measure. Most of the tests administered were good at distinguishing between brain-injured and nonbrain-injured people, although only two tests distinguished between the two brain-injured groups, i.e. those in PTA and those out of PTA. Almost all tests were good at monitoring recovery from PTA. PMID- 10462149 TI - A non-aversive rehabilitation approach for people with severe behavioural problems resulting from brain injury. AB - An approach is presented which uses exclusively non-aversive methods in the behavioural rehabilitation of people with severe behaviour problems resulting from acquired brain injury. The approach has five components: (1) behavioural assessment: analysing the way all aspects of a person's functioning may affect their behaviour, (2) positive programming: teaching the skills necessary to allow the person to achieve their desired ends without resorting to inappropriate behaviour, (3) ecological change: altering the environment to achieve a better match with the individual's cognitive deficits, (4) focused treatment: using behavioural contingencies to achieve a rapid reduction in target behaviour, and (5) reactive strategies: specifying action to be taken to gain short-term control over episodes of challenging behaviour. The current literature on behavioural rehabilitation is reviewed in the context of this approach. Two case studies are presented illustrating the use of the approach in practice. The strengths and potential pitfalls of the approach are discussed, along with issues which need to be considered for effective implementation. PMID- 10462150 TI - Characteristics and outcomes of a home and community-based neurorehabilitation programme. AB - The potential clinical and financial advantages of providing neurorehabilitation directly in patients' homes and communities have recently been discussed. However, the specific characteristics and outcomes of a coordinated, interdisciplinary, home-based programme does not currently exist in the rehabilitation literature. The present paper presents patient demographics, type and intensity of services provided, satisfaction measures, and clinical outcomes for 77 brain injured individuals in an attempt to begin to define and evaluate this new level of care. Additionally, the challenges of conducting home-based rehabilitation, and needs for further research are discussed. PMID- 10462151 TI - The persistence of mood disorders following traumatic brain injury: a 1 year follow-up. AB - The study investigated whether the frequency of mood disorders following traumatic brain injury (TBI) increased with time. Consecutive hospital admissions completed the Wimbledon Self-Report Scale at 6 and 12 months post-injury. Scores in the 'borderline/case' range were deemed clinically significant. Seventy-seven of those assessed at 6 months completed the 12 month follow-up. Those lost to follow-up (22) were not more likely to have been classified 'borderline/case' at 6 months. They were more likely to have been 'unoccupied' pre-injury (p=0.002). The frequency of clinically significant disorders did not change significantly, being 39% at 6 months and 35% 1 year post-injury. The majority of subjects (58) maintained the same clinical classification at both assessments. Eight of the 47 (17%) classified 'normal' at 6 months changed to 'borderline/case' at 12 months. However, 11 of the 30 (37%) classified 'borderline/case' at 6 months were 'normal' at 12 months. The direction of change was not statistically significant (p=0.5). These findings are consistent with previous research. However, the present study examined a larger sample and included a wider range of injury severity, thereby increasing the generalizability of the findings. Also, a broader spectrum of mood disorders was investigated which adds to previous work specifically on depression. PMID- 10462152 TI - Centiles of body mass index for Dutch children aged 0-20 years in 1980--a baseline to assess recent trends in obesity. AB - OBJECTIVE: To document the distribution of body mass index (BMI) in Holland in 1980 when obesity was uncommon. DESIGN: Nationally representative growth survey. SUBJECTS: Forty-one-thousand boys and girls age 0-20 years. METHOD: BMI centiles based on the original height and weight data were derived using the LMS method. RESULTS: Median BMI showed the familiar pattern of a rise in the first year, followed by a fall, then a second rise after 6 years. The level of overweight was less than for children from the USA, France and Norway measured at the same time or earlier. CONCLUSIONS: Dutch children in 1980 were relatively free of obesity, so the BMI centiles are a suitable baseline to monitor international trends in obesity since then. PMID- 10462153 TI - An analysis of variance of the pubertal and midgrowth spurts for length and width. AB - Using data from the first Zurich Longitudinal Growth Study characteristics of the growth of six variables--bihumeral width, biiliac width, standing height, sitting height, leg height and arm length--are studied. The main interest is in differences between boys and girls, and across variables and in particular in whether there are sex differences that are specific for some variables. For each child and variable, individual velocity and acceleration curves are estimated using a kernal smoother. From these curves, parameters characterizing the midgrowth spurt (MS) and the pubertal spurt (PS) are estimated: timings, durations and intensities. The level of childhood velocity is used for characterizing early growth. These parameters are analysed using a repeated measures analysis of variance (ANOVA) to assess the statistical significance of differences between boys and girls and across variables. This necessitates some kind of standardization and two types of standardization are used here. The MS shows negligible or small differences between boys and girls, and the same is true for velocity in childhood. Differences across variables during the MS are much more pronounced: with respect to intensity, bihumeral width has an MS about six times more intense than height. The PS is later for boys (as is well known), and there are significant differences across variables: bihumeral width and sitting height are late while legs are early. With the exception of biiliac width, the duration of the PS (which has been subdivided into three phases-early, middle and late) is slightly longer for boys for all variables: boys have a longer starting phase, the middle phase is about equal in length for both boys and girls, and girls have a slightly longer late phase. Leg height and height experience a PS of short duration while bihumeral and biiliac width experience a long one and these differences are highly statistically significant. For all variables, with the exception of biiliac width, boys have a more intense PS (in terms of maximal acceleration), even having adjusted for their larger adult size. Differences in intensity are also marked across variables, bihumeral width and sitting height having the highest intensity and legs the lowest. Differences between sexes and across variables are much smaller for the stopping intensity, characterized by maximal deceleration. PMID- 10462154 TI - Eight thousand years of economic and political history in Latin America revealed by anthropometry. AB - Human growth in height may be used as a cumulative record of the nutritional and health history of a person or a population, and often reflects the economic, social and political environment in which those people live. This paper explores the relationship between growth in height and the economic, social and political environment in Latin American populations. Adult height is analysed over an 8250 year period. It is shown that economic, social and political change prior to the European conquest of the Americas resulted in positive and negative trends in mean stature. Following the European conquest, there was a decline in mean adult stature in Middle and South America that continued until about 1939. From 1940 to 1989 there was a trend for increasing mean stature. A negative trend in stature for children is found in a second analysis. Economic decline and political unrest in Guatemala since 1978 is associated with a significant decline in the mean stature of 10- and 11-year-old children from families from very high, moderate, and very low socioeconomic status. PMID- 10462155 TI - A longitudinal study of the physical growth of Japanese infants. AB - In order to clarify the characteristics of the physical growth of Japanese infants up to 1 year of age, the data of 3183 infants from 53 hospitals in Japan were collected. Percentile values were calculated from the measurement of body weight, recumbent length, and head circumference, with some additional data interpolated to fill the gaps in the measurements. Compared to the National Center for Health Statistics (NCHS) standard, the recumbent length and head circumference of these infants were smaller throughout the observation period, while body weight was larger at 0-6 months of age, and smaller at 8-12 months of age. The cases were divided into five groups according to birth weight, to examine the influence of foetal growth. The 50th percentile curves of each group were parallel and the intervals between the 3rd and 97th percentile curves in each group became wider in the first 6 months of life, up to the interval of 2.20 3.18 Kg. There was no significant difference in body weight among three groups according to feeding methods, in any age period. PMID- 10462156 TI - Beta-globin haplotypes analysis in Afro-Brazilians from the Amazon region: evidence for a significant gene flow from Atlantic West Africa. AB - Beta-globin gene cluster haplotypes were analysed in betaA-, betaS- and betaC globin gene-bearing chromosomes in black people from Curiau, Pacoval and Trombetas, three communities made up of descendants of African slaves, located in the Northern region of Brazil. The betaA haplotype distribution is consistent with the African origin of the populations, with some degree of local differentiation and admixture with people of Caucasian ancestry and/or Amerindians. In addition, the betaS haplotype distribution (60% Bantu; 30% Senegal and 10% Benin) suggests that although African slaves brought to Northern region have been predominantly from regions where the Bantu haplotype predominates, there is also evidence of the presence of slaves from West Africa, particularly from the Atlantic West. PMID- 10462157 TI - Angiotensin-converting enzyme (ACE) alleles in the Quechua, a high altitude South American native population. AB - Recently it was reported that an allelic variant of the gene encoding angiotensin converting enzyme (ACE) was significantly over-represented in a cohort of elite British mountaineers. It was proposed that this may be evidence for a specific genetic factor influencing the human capacity for physical performance. The implication that this allele could enhance performance at high altitude prompted us to determine its frequency in Quechua speaking natives living at altitudes greater than 3000m on the Andean Altiplano in South America. We found that the frequency of the putative performance allele in the Quechuas, although significantly higher than in Caucasians, was not different from lowland Native American populations. This observation suggests that, although the higher frequency of the 'performance allele' may have facilitated the migration of the ancestral Quechua to the highlands, the ACE insertion allele has not been subsequently selected for in this high altitude population. PMID- 10462158 TI - Predictive parameters of joint disease in DBA/1 transgenic mice. AB - We recently reported an accelerated onset of collagen-induced arthritis in DBAII mice overexpressing a T cell receptor Valpha11.1/Vbeta8.2 transgene as a preclinical animal model for age-associated T cell dysfunction. The accelerated onset is due to a transgenically sensitized T cell population that reacts to bovine type 11 collagen without prior in vivo sensitization. The model presents a readily observable distal joint phenotype that would allow preliminary aging and intervention studies to be evaluated by monitoring the presence or absence or degree of phenotypic expression of disease. In order to characterize clinical signs, we evaluated 69 transgenic mice in six different experiments for anticollagen antibody levels, and assigned each a modified arthritic score based on the degree of redness or swelling of the digital joints. We also correlated these parameters with signs of distress, including weight bearing, activity levels, and body posture. The average onset of disease was consistently within a 28 to 35-day period. The average arthritic score at the time of onset was 8. We found that none of the parameters predicted the onset of joint disease, but the modified scoring system reflected the severity of joint disease and predicted the degree of distress associated with the acute inflammation. The ability to determine the severity of joint disease by gross physical examination is a useful clinical feature because a numerical score is reflective of the degree of inflammation. Because the transgenic mouse model is a T cell-driven disease, the effect of aging on T cell activity can be monitored easily. In addition, the use of our modified arthritic scoring system makes it possible to conduct mouse experiments in a humane manner. PMID- 10462159 TI - Aminoguanidine supplementation delays the onset of senescence in vitro in dermal fibroblast-like cells from senescence-accelerated mice. AB - The effects of aminoguanidine supplementation on senescence acceleration in vitro were examined in fibroblasts from the dorsal dermis of newborn SAMP11 (accelerated senescence-prone mice), and were compared to the effects in cell lines from SAMR1 (accelerated senescence-resistant) mice. Four millimolar aminoguanidine supplementation significantly delayed the senescence/crisis in cell lines from SAMP11 mice, but did not affect the senescence/crisis in cell lines from SAMR1 mice. Flow cytometric analysis of the DNA content of confluent cells revealed that aminoguanidine supplementation significantly decelerated the increase in the number of tetraploid cells until senescence/crisis. Although mean concentrations of lipid peroxides in the primary cultures did not differ significantly, considerably higher lipid peroxidation was observed in some SAMP11 cultures, and aminoguanidine supplementation reduced them to the levels in SAMRI cultures. These results strongly suggest that oxidative stress derived from polyamine catabolism may contribute to the senescence acceleration in vitro in cell lines from SAMP11 mice. PMID- 10462160 TI - Body fat distribution with long-term dietary restriction in adult male rhesus macaques. AB - Dietary restriction (DR) is the only intervention that has been shown to increase average and median life span in laboratory rodents. The effect of long-term, moderate DR on body composition and fat distribution was evaluated in male rhesus monkeys. Thirty animals (8-14 years of age)fed either 30% less than baseline intake (R, n = 15) or allowed to eat to satiety (C, n = 15), have been assessed semiannually using somatometrics and dual-energy alpha-ray absorptiometry (DXA)for 7.5 years. R subjects have reduced body weight (p <.0001), total body fat (p < .0001), and percentage body fat located in the abdominal region (p < .05). In addition, there has been a sustained reduction in plasma leptin concentrations (p <.001). These findings suggest reduced risk for common morbidities, such as insulin resistance, dyslipidemia, and type 2 diabetes mellitus, that are associated with advancing age and increased levels of bodyfat, especially in the visceral depot. PMID- 10462161 TI - The effects of peripheral vascular disease on gait. AB - This study was designed to determine whether patients with peripheral vascular disease (PVD) have gait abnormalities. A previous study on humans with PVD found no abnormalities whereas significant gait changes were seen with a rat model of PVD. The study population was comprised of 10 controls and 9 subjects with PVD (all male). The PVD group had documented pain in one or both legs while walking. Subjects ranged in age from 55-92 years of age, with a mean age of 69 in the PVD group and 70 in the control group. The GaitMat II system was used to measure both spatial and temporal variables of gait. Subjects walked across the mat, four to six times, at their comfortable walking speed. The PVD group then walked on a treadmill until they experienced moderate claudication pain and felt they had to stop (pain levels between 6 and 8, with maximal pain at level 10). Control group walked on a treadmill for 10 minutes without pain. All subjects repeated the gait tests on the GaitMat H system immediately after treadmill walking. Claudication pain persisted in the PVD group during the second gait test. The PVD group was not different than control group in any of the measured variables on the first test (p values from .35 to .99). Difference scores (post- minus pre-treadmill walking)for PVD group were significantly different than those for control group on 8 of 11 variables (p values <.005). The primary response in PVD subjects was reduced walking speed (1.02+/-0.16 to 0.94+/-0.16 m/s) and reduced step length (0.60+/-0.08 to 0.57+/-0.09 m/s), whereas control subjects increased their speed (1.09+/-0.17 to 1.19+/-0.19 m/s) and step length (0.63+/-0.10 to 0.67+/-0.10 m/s). No asymmetries in gait were measured in either group, either before or after treadmill walking. In conclusion, PVD subjects were not different in gait while rested, but responded to claudication pain by reducing preferred walking speed and step size. PMID- 10462162 TI - Commentary on "the effects of peripheral vascular disease on gait". PMID- 10462163 TI - Kleemeier award lecture: are there genes for aging? AB - The question of whether aging - the process that converts fit adults into frailer adults with a progressively increased risk of illness, injury, and death - is under genetic control is ambiguous, and its answer depends on what one means by aging. Natural selection can select for genes that retard aging, but only in species and niches where the value of prolonged survival outweighs its costs. Although the form aging takes can be affected by variations at many genetic loci the number of loci that moderate the pace of synchronized decay may be far smaller. Single gene mutants can extend mouse lifespan by over 50%, and genetic selection for small body size also leads to dramatic life span extension in dogs, suggesting strongly that aging can be affected by genetic variations within a species, but identification of genetic differences that discriminate long-lived from short-lived species will require a combination of genetic and physiological analyses. PMID- 10462164 TI - Spinal-flexibility-plus-aerobic versus aerobic-only training: effect of a randomized clinical trial on function in at-risk older adults. AB - BACKGROUND: As exercise is associated with favorable health outcomes, impaired older adults may benefit from specialized exercise interventions to achieve gains in function. The purpose of this study was to determine the added benefit of a spinal flexibility-plus-aerobic exercise intervention versus aerobic-only exercise on function among community-dwelling elders. METHODS: We employed a randomized clinical trial consisting of 3 months of supervised exercise followed by 6 months of home-based exercise with telephone follow-up. A total of 210 impaired males and females over age 64 enrolled in this study. Of these, 134 were randomly assigned to either spinal flexibility-plus-aerobic exercise or aerobic only exercise, with 116 individuals completing the study. Primary outcomes obtained at baseline, after 3 months of supervised exercise, and after 6 months of home-based exercise included: axial rotation, maximal oxygen uptake (VO2max); functional reach, timed-bed-mobility; and the Physical Function Scale (PhysFunction) of the Medical Outcomes Study SF-36. RESULTS: Differences between the two interventions were minimal. Overall change scores for both groups combined indicated significant improvement for: axial rotation (p=.001), VO2max (p=.0001), and PhysFunction (p=.0016). Secondary improvements were noted for overall health (p=.0025) and reduced symptoms (p=.0008). Differences between groups were significant only for VO2max (p=.0014) at 3 months with the aerobic only group improving twice as much in aerobic capacity as the spinal flexibility plus-aerobic group. Repeated measures indicated both groups improved during the supervised portion of the intervention but tended to return toward baseline following the home-based portion of the trial. CONCLUSIONS: Gains in physical functioning and perceived overall health are obtained with moderate aerobic exercise. No differential improvements were noted for the spinal flexibility-plus aerobic intervention. PMID- 10462165 TI - Reliability of foot trajectory measures within and between testing sessions. AB - BACKGROUND: Impaired control of foot trajectory during the swing phase of gait is hypothesized to increase the risk of slipping or tripping. Before assessing the predictive validity of foot trajectory measures with respect to incidence of falls, it is necessary to establish their reliability. The purpose of this study is to assess within- and between-session reliability of foot trajectory measures and traditional temporal-distance measures in healthy elderly women during gait. METHODS: Sixteen healthy, elderly women (ages 65-79 years) completed six sets of five trials each of natural and fast cadence gait during a 3.5-hour period on each of 4 days. An optoelectric motion analysis system and heel switches were used to obtain both foot trajectory (minimum toe clearance during swing, vertical, and horizontal heel contact velocities) and temporal-distance measures (step width, cadence, velocity, stride length, and time). RESULTS: Within-session test-retest reliability of all variables at natural and fast speeds was good to excellent, with intraclass correlation coefficients (ICCs) of greater than 0.9 for all but one measure (fast cadence stride time). ICCs for between-session test retest reliability were slightly lower, but still greater than 0.9 for all but two measures (fast cadence stride time and natural cadence vertical heel contact velocity). Heel contact velocities quantified at the instant of heel contact correlated strongly with values obtained by averaging over the last 2% of the gait cycle. DISCUSSION: The good to excellent within- and between-session reliability of these foot trajectory measures supports their use as a possible means of assessing subtle changes in gait motor control. Confirmation of an association between alterations in foot trajectory measures and incidence of falls awaits further study. PMID- 10462166 TI - Anxiety of Alzheimer's disease: prevalence, and comorbidity. AB - BACKGROUND: Anxiety may be associated with psychiatric morbidity, disability, increased health care utilization, and mortality in Alzheimer's disease (AD) patients as it is in the general adult population. However, the phenomenology of anxiety symptoms in AD and its relationship to dementia progression, comorbid depression, and the presence of other problematic behaviors have not yet been examined. METHOD: Data on anxiety symptoms and their coexistence with other factors were obtained in 523 community-dwelling AD patients through interviews with their caregivers and direct physical examination. The prevalence of anxiety symptoms and their association to patient depression, other behavioral problems, gender, and age was investigated. RESULTS: Anxiety symptoms were common, occurring in 70% of subjects. Anxiety symptoms were significantly correlated with ADL impairment and other behavioral disturbances, including wandering, sexual misconduct, hallucinations, verbal threats, and physical abuse. Comorbidity of anxiety-depression was also prevalent: 54% of the sample had both anxiety and depression symptoms. ADL impairment and problem behaviors were significantly associated with comorbidity; however, the latter association was explained entirely by the presence of anxiety. CONCLUSION: Anxiety symptoms were common and significantly related to ADL and additional neuropsychiatric problems in this sample. These results indicate the need for additional research into the phenomenology of anxiety and comorbid anxiety-depression in AD and for the development and investigation of effective assessment and treatment of anxiety in AD clinical practice. PMID- 10462167 TI - Peak aerobic power is an important component of physical performance in older women. AB - BACKGROUND: The purpose of this study was to determine the relationship between peak aerobic power (VO2peak) and performance on a modified Physical Performance Test (modified PPT) in older women. METHODS: One hundred one women aged 75 years and older seeking enrollment in randomized, controlled trials of exercise and/or hormone replacement therapy were recruited from the community-at-large and from congregate living sites. Measures obtained included VO2peak, a modified PPT, and self-reports about performance of activities of daily living. RESULTS: Simple regression analysis demonstrated that VO2peak was associated with total PPT score (r =.53, p <.001), gait speed (r =.44, p <.001), time to arise from a chair five times (r =.43, p = <.001), and time to climb one flight of stairs (r =.36, p =.007). Multiple regression analysis revealed that the relationships between VO2peak and total modified PPT score, gait speed, chair rise time, and time to climb one flight of stairs were independent of age. CONCLUSIONS: Peak aerobic power is a significant independent predictor of performance on a standardized test of physical function in older women and is an important component of physical frailty in this population. PMID- 10462168 TI - A high plasma concentration of TNF-alpha is associated with dementia in centenarians. AB - BACKGROUND: Inflammatory mechanisms and immune activation have been hypothesized to play a role in the pathogenesis of age-associated diseases such as dementia and atherosclerosis. The purpose of this study was to evaluate the plasma concentration of tumor necrosis factor (TNF)-alpha in a large cohort of centenarians and to look for its possible associations with cognitive function, atherosclerosis, and general health status. Furthermore, we investigated whether the concentration of TNF-alpha was correlated with the blood concentration of leucocyte subsets or the plasma concentrations of interleukin (IL)-6, soluble TNF receptor 11 (sTNFR-H) (75 kDa) and C-reactive protein (CRP). METHODS: Plasma TNF alpha was measured by ELISA in 126 centenarians, 45 subjects aged 81 years, 23 subjects aged 55-65 years, and 38 subjects aged 18-30 years. Atherosclerosis was evaluated by the ankle-brachial blood pressure index, and general health status was evaluated by the body mass index and the number of diagnoses present. RESULTS: The concentration of TNF-alpha was significantly increased in 126 centenarians compared to younger control groups, and a high concentration of TNF alpha was associated with both Alzheimer's disease and generalized atherosclerosis in the centenarians. The concentration of TNF-alpha was positively correlated with the concentrations of plasma IL-6, sTNFR-II, and CRP. No associations were found with increased leucocyte subsets or the body mass index. CONCLUSION: This study demonstrates that, even in apparently healthy subjects, age-associated immune activation indicated by raised levels of pro inflammatory cytokines may reflect age-associated pathological processes that develop over decades. PMID- 10462169 TI - Nursing home admission for African Americans with Alzheimer's disease. AB - BACKGROUND: For African Americans with Alzheimer's disease (AD), little is known about the time to, and risk factors for, nursing home admission (NHA). Using Consortium To Establish a Registry for Alzheimer's Disease (CERAD) data, this study provides information on NHA for African Americans. METHODS: This longitudinal study followed subjects (N=122) for as long as 7 years and used survival analysis methodology and variable values at baseline and at follow-up to identify NHA risk factors. Studied were sociodemographic variables, physical symptom and disease status variables, the Blessed Dementia Rating Scale (including subscores), the Clinical Dementia Rating (CDR), and the Mini-Mental State Examination. RESULTS: Only 25% of African Americans with AD were estimated to have had a NHA by 3.4 years (confidence interval 2.1, 5.4). Being unmarried resulted in a five times earlier NHA (p< .01), and each unit increase in the CDR resulted in a 74% earlier NHA (p<.01). In the absence of the CDR, limitation in activities of daily living was associated with earlier NHA (p<.05). CONCLUSIONS: Findings suggest that African Americans with AD spend a substantial time in the community prior to NHA, a longer time than observed in similar studies among whites. This raises public health and clinical concern that African Americans with AD may be residing in the community with substantial unmet needs, and that their caregivers have potentially high levels of burden. The independent associations with time to NHA observed here, although few in number, are consistent with other related research. PMID- 10462170 TI - Does religious attendance prolong survival? A six-year follow-up study of 3,968 older adults. AB - METHODS: A probability sample of 3,968 community-dwelling adults aged 64-101 years residing in the Piedmont of North Carolina was surveyed in 1986 as part of the Established Populations for the Epidemiologic Studies of the Elderly (EPESE) program of the National Institutes of Health. Attendance at religious services and a wide variety of sociodemographic and health variables were assessed at baseline. Vital status of members was then determined prospectively over the next 6 years (1986 1992). Time (days) to death or censoring in days was analyzed using a Cox proportional hazards regression model. RESULTS: During a median 6.3-year follow-up period, 1,777 subjects (29.7%) died. Of the subjects who attended religious services once a week or more in 1986 (frequent attenders), 22.9% died compared to 37.4% of those attending services less than once a week (infrequent attenders). The relative hazard (RH) of dying for frequent attenders was 46% less than for infrequent attenders (RH: 0.54, 95% CI 0.48-.0.61), an effect that was strongest in women (RH 0.51, CI 0.434).59) but also present in men (RH 0.63, 95% CI 0.52-0.75). When demographics, health conditions, social connections, and health practices were controlled, this effect remained significant for the entire sample (RH 0.72, 95% CI 0.64-.81), and for both women (RH 0.65, 95% CI 0.554-.76, p<.0001) and men (RH 0.83, 95% CI 0.69-1.00, p=.05). CONCLUSIONS: Older adults, particularly women, who attend religious services at least once a week appear to have a survival advantage over those attending services less frequently. PMID- 10462171 TI - The combined effects of baseline vulnerability and acute hospital events on the development of functional dependence among community-living older persons. AB - BACKGROUND: Many older persons who are highly vulnerable do not develop functional dependence, whereas some older persons with low vulnerability do develop functional dependence. We conducted this study to determine the combined effects of baseline vulnerability and precipitating events on the development of functional dependence. METHODS: We analyzed data from two prospective, population based cohort studies. The development cohort included 799 community-living persons, 72 years of age and older, who were independent in their activities of daily living (ADLs). The validation cohort included 1,051 comparable persons. Participants were classified by baseline vulnerability, defined on the basis of physical performance, cognitive status, and age, and by exposure to potential precipitating events, determined from information gathered from acute care hospital admissions. The primary outcome was the onset of functional dependence, defined as a new disability in one or more of the seven ADLs at the 1-year follow up interview or admission to a skilled nursing facility prior to the 1-year interview. RESULTS: Functional dependence developed in 109 (13.6%) participants in the development cohort and in 100 (9.3%) participants in the validation cohort. The rates of functional dependence for the low, intermediate, and high vulnerability groups were 7.1%, 17.2%, and 40.1% (p<.001) in the development cohort and 4.8%, 15.0%, and 28.0% (p<.001) in the validation cohort. For the four categories (none, mild, moderate, severe) of precipitating events, the rates of functional dependence were 9.0%, 19.4%, 27.3%, and 53.2% (p<.001 ) in the development cohort and 5.1%, 12.0%, 28.2%, and 53.3% (p<.001) in the validation cohort. For both cohorts, when baseline vulnerability and precipitating events were analyzed in cross-stratified format, the rate of functional dependence increased progressively from low-risk to high-risk groups in all directions (double-gradient phenomenon). The contributions of baseline vulnerability and precipitating events to the development of functional dependence were independent and statistically significant. CONCLUSIONS: Among community-living older persons, baseline vulnerability and precipitating hospital events contribute independently to the development of functional dependence and should each be targeted for intervention when developing strategies aimed at forestalling the onset of functional dependence. PMID- 10462172 TI - Retinol esterification activity contributes to retinol transport in stellate cells. AB - The mechanisms of retinol transport and accumulation in hepatic stellate cells (HSC) remain to be elucidated. Our previous studies suggested that retinol esterification activity, particularly lecithin:retinol acyltransferase (LRAT) activity, in liver retinoid metabolism is important to elucidate the relationship between retinol uptake by HSC and the esterification of retinol. In the present study, using a human HSC-like cell line, LI90, we demonstrated that retinol esterification activity of LI90 cells is similar to that of primary cultures of rat HSC and higher than that of a human hepatoma cell line. Further, since progesterone or diphospho-lauroyl-phosphatidylcholine increased retinol esterification activity of LI90 cells, it is likely that LRAT contributes to retinol esterification in LI90. We examined retinol esterification in LI90 cells and clearance of retinol from culture medium. The percentages of both retinol and esterified retinol in LI90 cells increased in a manner dependent on retinol concentration in medium, whereas that of retinol in medium decreased. The percentages of esterified and unesterified retinol in LI90 cells and of retinol in medium were linearly dependent on the logarithm of the initial concentration of retinol in the medium. These results suggest that retinol esterification activity contributes to retinol uptake by HSC and maintenance of non-toxic retinol levels in plasma. PMID- 10462173 TI - Isotype-specific changes in the amount of beta-tubulin RNA in synchronized tobacco BY2 cells. AB - The 3'-ends of the beta-tubulin cDNA were amplified from tobacco BY2 polyA+ RNA. According to the differences in the predicted amino acid sequence at the extreme C-terminal, they were grouped into three different isotypes, NTB1 in which "EEGDYYEEDEEDLNEA", NTB2 in which "EEEYYEDEEEA QED" and NTB3 in which "DECEYEEEEEYDHEGN" follows the conservative "YQQYQDATAD" sequence. Using unique 3'-untranslated regions as probes, changes in the RNA levels of each beta-tubulin isotype were determined by dot-blot hybridization. The levels exhibited characteristic rhythms in the cell cycle. NTB1 RNA was highest in S phase in comparison to NTB2 RNA level which was highest in late G2. On the other hand, NTB3 RNA level was highest in early G2. PMID- 10462174 TI - A new member of the GP138 multigene family implicated in cell interactions in Dictyostelium discoideum. AB - The cellular slime mold Dictyostelium discoideum reproduces sexually under submerged and dark conditions. Its mating system is polymorphic and particularly interesting with respect to mechanisms of cell recognition. The cell-surface glycoprotein gp138 has been implicated in sexual cell interactions, as it was identified as a target molecule for the antibodies that block sexual cell fusion in D. discoideum. Two mutually homologous genes, GP138A and GP138B, have been cloned, but gene disruption experiments to clarify their functional relationships suggested that there is at least one more gene for gp138. Further protein analysis including peptide mapping also revealed that gp138 exists as three isoforms, DdFRP1, DdFRP2, and DdFRP3. GP138A encodes DdFRP2 and GP138B, DdFRP3, and the presence of a third gp138 gene encoding DdFRP1 was suggested. Here, we isolated and characterized a third GP138 gene, GP138C. Although the deduced amino acid sequences of GP138C matched completely with those of peptide fragments of DdFRP1 in the N-terminal half, the rest did not give complete matches. Overexpression of GP138C caused an increase in the intensity of DdFRP1, but disruption of this gene did not diminish DdFRP1. Our results indicate that GP138C encodes a protein very similar to but distinct from DdFRP1. The GP138 multigene family is thus composed of more members than previously expected, and their functional relationships are of special interest. PMID- 10462175 TI - Effects of aluminum on plasma membrane as revealed by analysis of alkaline band formation in internodal cells of Chara corallina. AB - To study the mechanism of aluminum toxicity in plant cells, the effects of aluminum on alkaline band formation were analyzed in the internodal cells of Chara. After cells were treated with AlCl3, they were examined for their capacity to develop alkaline bands. Treating cells with AlCl3 medium at pH 4.5 completely inhibited alkaline band formation. When either CaCl2 or malic acid was added to the AlCl3 medium (pH 4.5), it did not produce an ameliorative effect, whereas addition of both CaCl2 and malic acid induced a significant ameliorative effect. It was found that treatment at pH 4.5 in the absence of AlCl3 strongly inhibited alkaline band formation. This inhibition by the low pH (4.5) treatment was effectively ameliorated by CaCl2. At higher pH (5.0), malic acid alone produced a significant ameliorative effect on aluminum inhibition of alkaline band formation, but CaCl2 did not. Recovery from aluminum inhibition was also studied. When cells treated with AlCl3 at pH 4.5 were incubated in artificial pond water, they could not recover the capacity to develop alkaline band. When either malic acid or CaCl2 was added to artificial pond water, cells recovered their alkaline band formation. It was concluded that one of the primary targets of aluminum is the plasma membrane and that aluminum affects the plasma membrane from the cell exterior at the beginning of the treatment (within 24 h). It was also suggested that the aluminum treatment impairs the HCO3- influx mechanism but not the OH- efflux mechanism. PMID- 10462176 TI - A cardiomyocyte mannose receptor system is involved in Trypanosoma cruzi invasion and is down-modulated after infection. AB - Mannosyl binding sites were detected "in vitro" on cardiomyocytes (CM) surface using horseradish peroxidase (HRP) as the ligand. Binding assays revealed a specific recognition system, which was time- and concentration-dependent. The binding required physiological pH and was inhibited by EDTA and trypsin treatments. HRP binding was reduced by pre-incubations with low concentrations of D-mannose. Ultrastructural analysis of the endocytic process was followed using HRP coupled to colloidal gold particles (HRP-Au). The tracer was found within caveolae characterizing early steps of the receptor-mediated endocytosis. The addition of 10 mM D-mannose to the interaction medium blocked Trypanosoma cruzi uptake by CM. The labeling of CM with a subsaturating concentration of HRP-Au before their infection showed, by ultrastructural studies, that its association with trypomastigote forms occurred frequently near to HRP-gold particles that could also be seen to comprise the parasitophorous vacuole. After infection of CM with T. cruzi, a considerable reduction on HRP binding was noticed. Binding was almost completely restored by treating the infected cultures with the trypanocidal drug Nifurtimox. Our "in vitro" findings suggest that cardiomyocyte's mannose receptors localized at the sarcolemma mediates T. cruzi recognition and can be down-modulated by parasite infection. PMID- 10462177 TI - Fas-induced in vivo apoptosis in bone marrow: anti-Fas mAb-induced elimination and successive proliferation of Fas-expressing cells especially those of myeloid lineage. AB - A single administration of agonistic anti-Fas mAb RK8 into mice decreased the number of bone marrow cells especially Mac1+ and Gr1+ cells of myeloid lineage. These cells, which were shown to be Fas-positive in normal bone marrow, were directly eliminated in vivo by Fas-mediated apoptosis. After the elimination of Fas-positive bone marrow cells, bone marrow was reconstituted by successive increase of numbers of Gr1(low) and Mac1(low) myeloid precursor cells expressing high levels of Fas, which are minor constituents in normal bone marrow. The increased cells consisted at least two components, Gr1(dull) Mac1+ cKit+ cells and Gr1(intermediate) Mac1+ cKit- cells, both of which were shown to be sensitive to Fas-induced apoptosis in vivo. Thus, Fas is functional in normal bone marrow and Fas-induced apoptosis in bone marrow enhances marked proliferation of Fas expressing myeloid precursor cells in vivo. PMID- 10462178 TI - Geranylgeranylacetone induces apoptosis in HL-60 cells. AB - Geranylgeranylacetone (GGA) induces apoptosis in human leukemia HL-60 cells in a dose- and time-dependent manner. This effect was completely prevented by the pan caspase inhibitor z-Val-Ala-Asp(OMe) fluoromethylketone, thereby implicating the caspase cascade in the process. Prior to DNA fragmentation, GGA treatment markedly activated caspase-3(-like) proteases, which might be responsible for the observed apoptosis. In addition, GGA treatment interfered with the processing and membrane localization of Rap1 and Ras, and these changes may be a result of apoptosis. Moreover, nitric oxide donors significantly accentuated the GGA induced apoptosis, suggesting that the apoptotic pathway induced by GGA might be regulated by a redox-sensitive mechanism. Taken together, these data suggest that the isoprenoid, GGA, is an effective inducer of apoptotic cell death in HL-60 cells. PMID- 10462179 TI - Effects of melatonin on neophobic responses in different strains of mice. AB - Anxiolytic properties of melatonin in rodents had usually been examined in behavioral tests based on stressful situations, i.e., in animal models of "state" anxiety. However, no study reports effects of melatonin on emotionality of rodents submitted to situations devoid of stressful components as in the free exploratory test, which gives to animals the opportunity to choose freely between familiar and unfamiliar places. This procedure has been proposed as a method for measuring an endogenous form of anxiety called "trait" anxiety. The present study first investigated the effects of melatonin on neophobic responses of male C57BL/6, C3H/He, and BALB/c mice submitted to a free-exploratory test. Results demonstrated that melatonin had no effect in C57BL/6 mice that presented very low neophobic responses, whereas it was effective in reducing neophobia of BALB/c and C3H/He mice that presented, respectively, strong and intermediate avoidance responses towards unfamiliarity. Indeed, mice of both latter strains treated with melatonin made fewer attempts to enter into the unfamiliar compartment, exhibited a lower latency of the first entry into the unfamiliar places, and spent more time in them. Thus, melatonin appeared to be equally effective in reducing "trait" anxiety in both BALB/c and C3H/He mice. Moreover, flumazenil was able to counteract, in a dose-dependent manner, the anxiolytic activity of melatonin in BALB/c, suggesting involvement of central GABAergic system in the pharmacological effects of melatonin. PMID- 10462180 TI - Dissociation of (-) baclofen-induced effects on the tail withdrawal and hindlimb flexor reflexes of chronic spinal rats. AB - We previously reported that the antinociceptive effect of the GABA(B) receptor agonist, (-)baclofen, in chronic spinal rats depended on the route of administration. That is, subcutaneous (SC) injections significantly increased the latency of the thermally elicited tail withdrawal (tail flick, TF) reflex, whereas spinal (intrathecal, IT) injections did not. The present studies attempted to determine the reason for this differential response. The possible contribution of a peripheral component to the systemic effect was evaluated, but was not supported by negative results of intradermal (-)baclofen injections (50 and 500 microg) into the tail skin of chronic spinal rats. A spinal site of action was indicated when pretreatment with 30 microg, IT of the GABA(B) receptor antagonist, phaclofen, significantly reduced the antinociceptive effect of SC ( )baclofen in both chronic spinal (5 mg/kg) and intact rats (2 mg/kg). Moreover, direct IT injections of (-)baclofen in chronic spinal rats produced a modest, but statistically significant increase in TF latency at doses of 0.06, 0.12, 0.3, and 0.6 microg, but not 1.2 microg. In the same spinal preparation, the flexor response was significantly reduced by IT injection of 0.6 and 1.2 microg, but not lower doses of 0.3 and 0.12 microg. These results provide the first quantitative, electrophysiological evidence of an antispastic effect of IT (-)baclofen in an in vivo, unanesthetized animal model. Second, the data show a separation between an antinociceptive effect of low spinal doses and an antispastic/muscle relaxant effect at higher doses, which may account for the results of our prior report. Finally, the data are also consistent with behavioral reports of antiallodynic/analgesic effects of low-dose baclofen, and may be relevant to the electrophysiological evidence of a preferential presynaptic action of low-dose ( )baclofen at the primary afferent synapse. PMID- 10462181 TI - Effects of perospirone, a novel 5-HT2 and D2 receptor antagonist, on Fos protein expression in the rat forebrain. AB - The effects of perospirone, a novel 5-HT2 and D2 receptor antagonist, on Fos protein expression in the nucleus accumbens (NA) and dorsolateral striatum (DLSt) were compared with those of typical (i.e., haloperidol and fluphenazine) and atypical (i.e., clozapine and risperidone) antipsychotics using immunohistochemical techniques in rats. Perospirone and other antipsychotics tested at doses that exerted D2 blocking actions increased Fos-like immunoreactivity both in the NA and DLSt. However, the levels of Fos expression in the DLSt induced by perospirone and clozapine were less than those induced by haloperidol and fluphenazine. When compared the differences in numbers of Fos positive neurons between in the NA and DLSt, perospirone, clozapine, and risperidone preferentially increased Fos expression in the NA. These findings suggest that perospirone has a preferential action on the mesolimbic (vs. nigrostriatal) dopaminergic system in inducing Fos protein in the rat brain, which may be related to its atypical antipsychotic properties. PMID- 10462182 TI - Context-independent sensitization to the locomotor-activating effects of cocaine. AB - After repeated intermittent exposure to psychostimulants, an increase in the behavioral response to the drug is observed. The development of this sensitized response is greatly influenced by environmental cues. For example, when the pretreatments are administered in an environment distinct from the test, a sensitized response is often not observed. This finding has led some investigators to suggest that sensitization is completely context dependent. The present experiment established context-independent sensitization by administering pretreatments in an environment distinct from the test and measured the effects of pretreatment on potency and/or efficacy of subsequent cocaine administrations. Separate groups of rats received single or multiple daily injections of cocaine (10.0 mg/kg) or the saline vehicle in the home cage during a 5-day pretreatment phase. Ninety-six hours following the last of the pretreatment injections the locomotor-activating effects of cocaine (0.0, 5.0, 10.0, or 20.0 mg/kg) were measured. For control rats, a significant increase in motor activity was obtained following administration of the 20.0 mg/kg dose. Rats that received the cocaine pre-treatment became sensitized to cocaine's motor activating effects. For these rats, cocaine pretreatment produced a leftward shift in the dose-effect curve, consistent with an increased potency. The maximum locomotor response was not altered by pretreatment, suggesting that drug efficacy was not effected by preexposure. Thus, context-independent sensitization to cocaine reflects an increased potency, but not efficacy, of the drug. PMID- 10462183 TI - Effects of arginine-vasopressin fragment 4-9 on rodent cholinergic systems. AB - Arginine-vasopressin fragment 4-9 (AVP4-9) has been demonstrated in animal studies to facilitate learning and memory. To clarify the mechanisms of this facilitation, we focused on the effects of AVP4-9 on rodent cholinergic systems. AVP4-9 (0.1 microM) enhanced the basal and the high-potassium-evoked acetylcholine (ACh) release from rat hippocampal slices (122.4 and 120.0% of control, respectively) in the presence of 1.3 mM calcium (physiological level) at 60 min after the incubation at 37 degrees C. The AVP4-9-stimulated basal ACh release was inhibited by a V1-selective antagonist ([(beta-mercapto-beta,beta cyclopentamethylene propionic acid)1, O-methyl-Tyr2, Arg8] vasopressin), but not by a V2-selective antagonist ([adamantaneacetyl1, O-ethyl-D-Tyr2, Val4, aminobutyryl6, Arg8,9]-vasopressin). In addition, AVP4-9 did not affect the basal ACh release under the calcium-free condition at 37 degrees C or in the presence of 1.3 mM calcium at 4 degrees C. However, AVP4-9 facilitated the passive avoidance response of scopolamine (a cholinergic blocker)-induced memory deficient mice. These findings demonstrate that AVP4-9 stimulates ACh release via mediation by V1-like vasopressin receptors, and shows dependence on calcium ion and temperature. The results also suggest that the mechanism of the facilitative effects of AVP4-9 on learning and memory consist of the observed stimulation of cholinergic systems and other parallel pathways that would not be inhibited by cholinergic blocking. PMID- 10462184 TI - Contribution of glutamatergic systems in locus coeruleus to nucleus paragigantocellularis stimulation-evoked behavior. AB - The role of extracellular glutamate, within the locus coeruleus, in mediation of the behavioral signs elicited by electrical stimulation of the nucleus paragigantocellularis (PGi) was investigated in conscious, opioid-naive rats. Each rat was prepared with a chronically implanted unilateral electrode within the PGi and a microdialysis guide cannula directed at the ipsilateral locus coeruleus. Opioid withdrawal-like behaviors (rearing, teeth-chattering, wet-dog shakes, etc.) and increases in extracellular glutamate concentrations within the locus coeruleus were evoked, in a frequency-dependent (0.5-50 Hz) manner, during PGi stimulation. Reverse dialysis perfusion of the locus coeruleus with the nonspecific glutamate receptor antagonist, kynurenic acid (0.1, 1 mM), reduced the intensity of stimulation-induced behaviors by roughly 50%, but had no effect on the corresponding increases in glutamate concentrations. Perfusion of the locus coeruleus with the glutamate transporter inhibitor, L-trans-pyrrolidine dicarboxylic acid, at 1, but not at 0.1, mM significantly increased glutamate levels in dialysates. Neither concentration of the transporter inhibitor altered the behavioral score. The results indicate that the opioid withdrawal-like behaviors elicited by electrical stimulation of the brainstem at the site of the PGi are positively correlated with locus coeruleus levels of glutamate, and suggest further that the behaviors are partially mediated by release of glutamate within the locus coeruleus or its immediate vicinity. PMID- 10462185 TI - Effects of calcium channel blockers on behaviors induced by the N-methyl-D aspartate receptor antagonist, dizocilpine, in rats. AB - The present study assessed the ability of voltage-sensitive calcium channel (VSCC) blockers to affect the behavioral effects of the noncompetitive N-methyl-D aspartate (NMDA) receptor antagonist, dizocilpine, in male Wistar rats. Dizocilpine produced dose-dependent increases in locomotor activity. Nimodipine, verapamil, and flunarizine suppressed dizocilpine-facilitated vertical activity, while horizontal activity was attenuated by verapamil and nimodipine but not flunarizine. Repeated dizocilpine injections resulted in the development of sensitization to its locomotor stimulating properties. Development of sensitization was not context specific, and was observed following repeated exposures to 0.1 but not 0.056 or 0.3 mg/kg of dizocilpine. Nimodipine retarded the development of sensitization to dizocilpine's stimulating effects on horizontal activity, while verapamil suppressed sensitization to the vertical stimulating effects of dizocilpine. Flunarizine had no significant effects on sensitization to dizocilpine's locomotor stimulating properties. In rats trained to discriminate between injections of 0.056 mg/kg of dizocilpine and vehicle, none of the tested VSCC blockers was able to completely antagonize the discriminative stimulus properties of dizocilpine. Nimodipine, when administered in combination with the training dose of dizocilpine, modestly decreased the dizocilpine-lever selection. Dizocilpine dose dependently decreased the self determined stimulation threshold implanted in rats with electrodes into the ventral tegmental area. Nimodipine exhibited some tendency to block the facilitating effects of dizocilpine, while verapamil and flunarizine had no effects. In summary, in the present experiments VSCC blockers exerted only modest interactions with the behavioral effects of dizocilpine, and it is unlikely that VSCC blockers have remarkable potential as adjunct treatment aimed at correcting the negative side effects of NMDA receptor antagonists (e.g., dizocilpine). PMID- 10462186 TI - Extracellular glutamate in the dorsal horn of the lumbar spinal cord in the freely moving rat during hindlimb stepping. AB - The capacity to reestablish locomotor function after complete spinal cord transection in the adult mammal is now well documented. Further studies have shown different neurotransmitters to be involved in the initiation and maintenance of these locomotor patterns. However, there has been no in vivo evidence of the changes in glutamate or any other neurotransmitter in the extracellular space of the dorsal horn during an alternating motor pattern such as hindlimb stepping. This study describes an in vivo microdialysis technique to measure extracellular glutamate in the dorsal horn of the spinal cord in the fully awake intact rat. A concentric microdialysis probe was placed in the dorsal horn at L5, and 18 h later dialysate samples were collected at 20-min intervals before, during, and after 20 min of hindlimb stepping. During stepping, extracellular glutamate rose 150% above resting levels and returned to resting levels 40 min later. This increase may have occurred either as a result of primary afferent depolarization or modulation by the descending and ascending supraspinal pathways. In another series of experiments extracellular glutamate was, therefore, measured in the dorsal horn of the chronic spinally transected rat during 20 min of hindlimb stepping. Although the spinal group did not take as many steps as the intact group, those taking more than 40 steps showed a significant rise in extracellular glutamate, and the number of steps taken by the individual spinal rats correlated positively with the individual values of extracellular glutamate (r2 = 0.63). These results are consistent with glutamate being an important neurotransmitter in the spinal cord in normal locomotion. PMID- 10462187 TI - Caffeine and stroop interference. AB - In two experiments the hypothesis that caffeine reduces Stroop interference was tested. In the first experiment interference was measured as the reduction in performance when subjects had to indicate the numerosity of strings of incongruent digits, relative to neutral-symbol strings. In the second experiment the incongruent condition consisted of naming the color of words referring to incongruent colors, and was compared to color naming of neutral strings. In the number-digit task 250 mg caffeine reduced interference at the level of error rates, relative to placebo. In the color-word task interference was reduced at the level of reaction times. These results were obtained with blocked presentations of incongruent and neutral conditions. The color-word experiment also contained a condition in which neutral and incongruent trials were mixed within one sequence. In this mixed condition caffeine still reduced overall reaction times, but no longer specifically interference. It is argued that this dissociation reflects a caffeine-induced increase in flexibility. The results are discussed in relation to failed previous attempts to demonstrate increased selectivity under caffeine using non-Stroop tasks, the importance of including pretreatment sessions to detect artificial effects, and the possible contribution of withdrawal effects. PMID- 10462188 TI - An investigation of serotonergic involvement in the regulation of ACTH and corticosterone in the olfactory bulbectomized rat. AB - The bilateral olfactory bulbectomy resulted in significantly higher plasma concentration of corticosterone, but not of ACTH in basal conditions and much higher plasma ACTH and corticosterone concentrations after 15 min of immobilization stress than were observed in sham-operated animals. Daily treatment with fluoxetine-a specific serotonin reuptake inhibitor-(15 mg/kg/day) had no effect on basal ACTH and corticosterone concentrations in OB rats. Fluoxetine treatment caused lower levels of ACTH, but not of corticosterone secretion, in response to immobilization stress. Bulbectomy significantly reducing 5-HT concentration in the amygdala. Stress increased serotonergic activity in the hypothalamus but not in the amygdala of OB rats. Chronic fluoxetine treatment of both unstressed and stressed OB rats resulted in a lower turnover rate in the two structures. Our results suggest that the hypercorticosteronemia observed after bulbectomy in unstressed OB rats is independent of the serotonergic system in both hypothalamus and amygdala. In contrast, they also demonstrate hypothalamic 5-HT changes in the HPA hyperactivity of OB rats in response to stress. Chronic fluoxetine treatment may normalize pituitary ACTH secretion in response to stress, possibly desensitization of the 5-HT receptors in the hypothalamus due to 5-HT being move available at the synapses. PMID- 10462189 TI - Lack of tolerance to ethanol-induced motor impairment on accelerod performance in rats. AB - The effect of ethanol on rats was investigated at increasing rates of acceleration for bar rotation speed. Ethanol was given to rats by a liquid diet starting with 2.4% ethanol (v/v) for 3 days. Then the ethanol concentration was increased to 4.8% (v/v) for 3 days and finally to 7.2% (v/v) for 15 days. Accelerod performance was recorded before and throughout 20 days of ethanol intake. Mean blood ethanol levels were 266.34+/-13.11 and 285.20+/-9.77 mg/dl on the 7th and 15th days of ethanol (7.2% v/v) consumption, respectively, as measured in a parallel group of animals. Ethanol produced significant concentration-dependent impairments in the accelerod performance of rats. The motor impairment effect of ethanol was most prominent in the test using the greatest rate of acceleration (from 0 to 79 rpm within 2 min). The impairment effect of ethanol on accelerod performance occurred throughout the period of ethanol exposure. Our results indicate that motor impairment on the accelerod performance test produced by an ethanol liquid diet depends on the concentration of ethanol and the rate of acceleration. In addition, under free-access conditions accelerod performance may not be a suitable behavioral test for detecting tolerance development to ethanol in rats. PMID- 10462190 TI - Morphine tolerance and dependence in mice with history of repeated exposures to NMDA receptor channel blockers. AB - Mice were subjected to two successive treatment protocols: first with NMDA receptor channel blockers (14 days, once a day) and second with morphine (5 mg/kg, 8 days, once a day). Treatment with the higher doses of dizocilpine (1 mg/kg), memantine (30 mg/kg), and MRZ 2/576 (30 mg/kg) upon discontinuation revealed only minor behavioral abnormalities attributable to the state of withdrawal. Following repeated administration of low-dose morphine, tolerance to morphine analgesia developed in mice preexposed to dizocilpine (1 mg/kg but not 0.3 mg/kg) but not memantine (10 and 30 mg/kg), MRZ 2/579 (10 and 30 mg/kg), or saline. There were no signs of morphine dependence in any treatment group. Overall, the present study found only minor effects of the subchronic administration of high doses of NMDA receptor channel blockers, suggesting that clinical use of NMDA receptor channel blockers such as memantine will not be accompanied by increased propensity to induction of morphine tolerance and dependence. PMID- 10462191 TI - Anticonvulsant activity and plasma level of 2,3-benzodiazepin-4-ones (CFMs) in genetically epilepsy-prone rats. AB - Anticonvulsant properties of some 2,3-benzodiazepine derivatives acting as alpha amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) antagonists have been examined in vivo in the genetically epilepsy-prone rats using an audiogenic seizures assay. 2,3-Benzodiazepin-4-ones (CFMs) are nonselective AMPA antagonists that have been found to be potent anticonvulsant compound is in acute models of epilepsy. Because very little is known about their actions in a chronic model of epilepsy, and no correlations exist between anticonvulsant potency and plasma levels of these derivatives, we planned to investigate such a relationship. Maximal anticonvulsant protection occurred 15-60 min after the IP administration of GYKI 52466, 30-90 min after CFM-2, and 45-120 min after CFM-3. In addition, maximal anticonvulsant effect was observed 60-120 min after the IP administration of CFM-4 and at 90 min after CFM-5. The therapeutic index revealed that GYKI 52466 was slightly more toxic than CFM-2 and CFM-3. The time course of plasma levels of rats treated showed that peak plasma concentration was observed 45 min after IP administration of CFM-2 and CFM-3 and 75 min after CFM-4 and CFM-5. Following IP administration of CFM-3 two curves were detected, one is referred to the injected compound, and the other to its demethylated metabolite, which corresponds to CFM-2. Also. for the nitroderivative CFM-4 two curves were detected: one of an injected compound and the second due to its reduced metabolite (CFM-2). Finally, three different metabolites were detected in rat plasma after IP administration of CFM-5. The present study demonstrated that CFMs showed a significant protection against auditory stimulation during the period of peak plasma concentrations, suggesting a marked inhibition of those brain structures involved in the initiation and/or spreading of the audiogenic seizures. PMID- 10462192 TI - Endotoxin- and interleukin-1-induced hypophagia are not affected by adrenergic, dopaminergic, histaminergic, or muscarinic antagonists. AB - Endotoxin (lipopolysaccharide, LPS) and interleukin-1 (IL-1) administration induce hypophagia in rodents. Both IL-1 and LPS are known to activate cerebral norepinephrine and serotonin metabolism, and IL-1 affects that of acetylcholine and histamine. Each of these neurotransmitters has been implicated in feeding behavior. Therefore, the ability of specific antagonists of the above neurotransmitter systems to counteract feeding responses to peripherally injected mIL-1beta and LPS was studied. Feeding was assessed in nondeprived mice by measuring the intake of sweetened milk in a 30-min period, as well as daily food pellet intake. LPS and mIL-1beta reliably reduced milk intake, and often reduced food pellet intake and body weight. Treatment of the mice with peripherally administered alpha-adrenergic (phentolamine or prazosin) or 3-adrenergic antagonists (propranolol), either alone or in combination, did not significantly alter the hypophagic responses to mIL-1beta or LPS. Mice in which cerebral norepinephrine was depleted with DSP-4 or 6-hydroxydopamine also displayed the usual hypophagia in response to mIL-1beta and LPS. The hypophagic responses to mIL-1beta and LPS were not affected by the histaminergic antagonists, pyrilamine (H1), cimetidine (H2), thioperamide (H3), or the histamine-depleting agent, alpha fluoromethylhistidine, nor by the muscarinic cholinergic antagonist, scopolamine. The responses to mIL-l1 were also unaffected by the dopamine receptor antagonist, haloperidol, the opioid receptor antagonist, naloxone, and the NO synthase inhibitor, L-NAME. These results suggest that adrenergic, dopaminergic, histaminergic, cholinergic, opioid or nitric oxide systems are not essential for the hypophagia induced by IL-1, and that multiple redundant pathways may be involved in illness-related hypophagia. PMID- 10462193 TI - The effects of neurosteroids on rat behavior and 3H-muscimol binding in the brain. AB - The effects of ICV administration of metabolites of progesterone and deoxycorticosterone [i.e., neurosteroids: AP (3alpha-hydroxy-5alpha-pregnan-20 one, allopregnanolone), 5alpha(-THDOC (3alphat-21-dihydroxy-5alpha-pregnan-20 one, 5alpha-tetrahydrodeoxycorticosterone), 5beta-THDOC (3alpha-21-dihydroxy 5beta-pregnan-20-one, 5beta-tetrahydrodeoxycorticosterone), and PS (3beta-hydroxy 5-pregnen-20-one sulfate, pregnenolone sulfate] were studied in the open-field test of neophobia and Vogel's test of conflict behavior in rats. The influence of in vivo administered 5beta-THDOC, a positive allosteric modulator of the GABA(A) receptor complex, on 3H-muscimol binding in different brain structures, was also studied with the help of quantitative autoradiography. The presented data did not reveal any anxioselective effects for a range of centrally active neurosteroids, in the ethologically orientated and conflict models of anxiety, after intracerebral drug administration. Their central effects appeared secondary to changes in rat gross behavior. It is possible that high local concentration of neurosteroids after ICV injection and production of a narrower range of behavioral effects than that of benzodiazepines, precluded manifestation of the antianxiety effects of AP, 5alpha-THDOC and 5beta-THDOC. Autoradiography did not reveal any significant changes in the specific binding of 3H-muscimol in brain structures after in vivo ICV administration of 5beta-THDOC at the behaviorally active dose. Thus, the possibility that neuroactive neurosteroids may provide a novel potential site for therapeutic interventions in anxiety disorders is not supported. The part of the experiment with 5beta-THDOC is interpreted as contributing to other results, suggesting the existence of a new category of neurosteroids acting as partial agonists of the GABA(A) receptor. PMID- 10462194 TI - Alnespirone (S 20499), an agonist of 5-HT1A receptors, and imipramine have similar activity in a chronic mild stress model of depression. AB - A chronic mild stress (CMS) model of depression was used to study an antidepressant-like activity of alnespirone (S 20499), a selective agonist of 5 HT1A receptors. In this model, a substantial decrease in consumption of a palatable sucrose solution over time is observed in rats subjected to a variety of mild stressors. This effect can be reversed by chronic administration of various classes of antidepressant drugs. Chronic (5 weeks) treatment with alnespirone, in a dose range between 1-5 mg/kg/day, gradually and dose dependently reversed the CMS-induced reductions in sucrose consumption without any significant effects in the non-stressed control animals. The onset of action of the most active doses (2.5 and 5 mg/kg/day) and the overall efficacy of alnespirone in the CMS model were comparable to those observed following similar administration of imipramine (10 mg/kg/ day). At the lower (0.5 mg/kg/day) and higher (10 and 20 mg/kg/day) doses, alnespirone was ineffective against the CMS induced deficit in sucrose consumption. These data provide further support for previous suggestions, based on both the clinical observations and animal data, that agonism at 5-HT1A receptors may result in antidepressant action. PMID- 10462195 TI - Scopolamine inhibition of female sexual behavior in rhesus monkeys (Macaca mulatta). AB - Previous research supports the activational role of central cholinergic mechanisms in rodent female sexual behavior. This experiment examined if similar central cholinergic mechanisms facilitate female rhesus monkey (Macaca mulatta) sexual behavior. Eight ovariectomized female rhesus monkeys received daily estradiol benzoate priming (5 microg/kg, SC). After 13-16 days of estrogen priming, animals were injected intravenously with either the cholinergic antagonist, scopolamine (0.70 mg/kg), or saline vehicle (1 ml/kg). Results indicate that the female proceptive behaviors of noncontact presentations significantly decreased 1545 min after scopolamine injection. Scopolamine inhibition was sustained up to 75 min only after 15 days of estrogen priming. Scopolamine did not significantly reduce other female sexual behaviors. Additionally, significant decreases in the number of mounts and intromissions, but not hip touches, were displayed by males exposed to scopolamine-treated females. This research suggests the possibility of a central cholinergic mechanism regulating female sexual behavior in rhesus monkeys. However, the general nature and duration of the cholinergic regulation of primate female sexual behavior differs substantially when compared to rodent behavior. PMID- 10462196 TI - Facilitation of memory performance by a novel muscarinic agonist in young and old rats. AB - AF150(S), a partial M1 muscarinic receptor agonist, was tested for its ability to improve performance in rats in a delayed matching-to-position task. Young and old rats received intraperitoneal injections of 0, 1, and 4 mg/kg of AF150(S). AF150(S) significantly enhanced matching accuracy for both young and old rats. Fits of exponential functions to discriminability measures showed that the enhancement was manifest as a reduction in the rate of forgetting. These results indicate that AF150(S) may be a useful therapeutic agent for improving cognitive function. PMID- 10462197 TI - An in vitro model for screening oral hypoglycemics. PMID- 10462198 TI - Aged garlic extract attenuates the cytotoxicity of beta-amyloid on undifferentiated PC12 cells. PMID- 10462199 TI - Cell-specific activation of the HB-EGF and ErbB1 genes by stretch in primary human bladder cells. PMID- 10462200 TI - Selective cytotoxicity of 3-amino-L-tyrosine correlates with peroxidase activity. AB - In the presence of 3-amino-L-tyrosine (3-AT), abundant brown pigment forms in human HL-60 cells, but not in a variety of other cell lines, which are reported to be lower in mean myeloperoxidase (MPO) content than HL-60. Cells were assessed for peroxidase activity with an ABTS-based colorimetric assay and compared to values obtained with known amounts of human myeloperoxidase. HL-60 cells were estimated to contain the equivalent of 37.1 ng myeloperoxidase/10(6) cells versus 26.1 and 5.0 ng/10(6) cells for human K562 and murine RAW 264.7 cell lines, respectively. HL-60 cells exhibited a nearly 60% inhibition of proliferation and > 70% reduction in cell viability after 4 d of culture in the presence of 100 microg 3-AT per ml. Higher concentrations of 3-AT (up to 400 microg/ml) for 4 d reduced HL-60 proliferation by 80% and decreased viability to 1-3%. Comparable levels of cytotoxicity were achieved in KG-1 cells after 7 d with 200 or 400 microg 3-AT per ml. K562 cells exhibited a 40% reduction in cell number after 7 d with 400 microg 3-AT per ml, but concentrations less than 400 microg/ml did not significantly affect K562 proliferation. K562 viability remained unchanged with doses of 3-AT up to 400 microg/ml. RAW 264.7 cells exhibited unchanged viability and proliferation in the presence of 3-AT at concentrations up to 400 microg 3-AT per ml. K562, KG-1, and RAW 264.7 cells exhibited no evidence of brown pigment formation in the presence of 3-AT and medium containing 10% fetal bovine serum. However, RAW 264.7 cells that were converted to protein-free medium and exposed to 3-AT exhibited intense brown pigment in some cell nuclei. A high percentage of HL-60 cells treated with 3-AT exhibited membrane blebbing, pyknosis, and nuclear fragmentation, which was not observed among other 3-AT-treated cell lines. A mechanism involving toxic intermediates of peroxidase-mediated "aminomelanin" formation is hypothesized. PMID- 10462201 TI - Relationship between bisphosphonate concentration and osteoclast activity and viability. AB - Difluoromethylidene bisphosphonate (F2MBP) is one of the many bisphosphonates known to inhibit bone resorption in vitro and in vivo. We have developed an analytical method, employing anion exchange and postcolumn indirect fluorescence detection, by which F2MBP can be quantified in bone samples. The objective of this study was to relate the concentration of F2MBP in embryonic bones treated in organ culture to the physiological effects of the compound, such as bone resorption (i.e., the amount of 45Ca released into the medium from prelabeled bones) and viability of the osteoclast population (i.e., the incidence of abnormal osteoclasts). Osteoclasts in bones treated with F2MBP exhibited morphological features of apoptosis, such as nuclear fragmentation. Both the number and percentage of these abnormal cells increased with dose of F2MBP and duration of incubation. The decrease in normal osteoclasts was correlated with the decreased amount of 45Ca released into the medium. Bones treated with F2MBP for only the first 5 min of the 48-h incubation period had similar numbers of abnormal osteoclasts and amounts of 45Ca released, as had bones incubated with F2MBP continuously for 48 h. The uptake of F2MBP into the bone was rapid. Bones treated with F2MBP for 6 h were similar to bones treated with F2MBP for the entire 48-h incubation period, both in F2MBP concentration and the 45Ca release ratios. These relationships between concentrations of F2MBP within bone and osteoclast activity and viability implicate apoptosis in the mechanism by which this bisphosphonate inhibits bone resorption. PMID- 10462202 TI - Establishment and characterization of 13 cell lines from a green turtle (Chelonia mydas) with fibropapillomas. AB - Thirteen cell lines were established and characterized from brain, kidney, lung, spleen, heart, liver, gall bladder, urinary bladder, pancreas, testis, skin, and periorbital and tumor tissues of an immature male green turtle (Chelonia mydas) with fibropapillomas. Cell lines were optimally maintained at 30 degrees C in RPMI 1640 medium supplemented with 10% fetal bovine serum. Propagation of the turtle cell lines was serum dependent, and plating efficiencies ranged from 13 to 37%. The cell lines, which have been subcultivated more than 20 times, had a doubling time of approximately 30 to 36 h. When tested for their sensitivity to several fish viruses, most of the cell lines were susceptible to a rhabdovirus, spring viremia carp virus, but refractory to channel catfish virus (a herpesvirus), infectious pancreatic necrosis virus (a birnavirus), and two other fish rhabdoviruses, infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus. During in vitro subcultivation, tumor-like cell aggregates appeared in cell lines derived from lungs, testis, and periorbital and tumor tissues, and small, naked intranuclear virus particles were detected by thin section electron microscopy. These cell lines are currently being used in attempts to isolate the putative etiologic virus of green turtle fibropapilloma. PMID- 10462203 TI - Quantitative aspects of the selective killing of transformed cells by methotrexate in the presence of leucovorin. AB - A quantitative study was made of the cytotoxicity of methotrexate (MTX) for nontransformed and transformed NIH 3T3 cells in the presence and absence of leucovorin. The study was preceded by an analysis of the growth rates of the cells at low and high population density combined with low and high concentrations of calf serum (CS). The reduced maximal growth rates of the transformed cells at low population densities relative to the nontransformed cells reinforced earlier evidence that heritable damage involving chromosome aberrations drives the process of transformation. When small numbers of transformed cells are cocultured with a large excess of nontransformed cells in the assay for transformed foci, the transformed cells were more readily killed by MTX than the nontransformed cells. The selectivity was increased when leucovorin (folinic acid) was present in the medium. The selective killing of the transformed cells actively multiplying in foci was most pronounced when the background of nontransformed cells had become confluent and their growth was inhibited. However, selectivity has also been demonstrated when transformed and nontransformed cells are growing at their maximum rates at low density despite the lower growth rate of the transformed cells under these conditions. The sensitivity of transformed cells in pure culture to MTX was lower during the first 3 d of subculture than in the following 6 d but decreased to zero a few d after net growth had ceased. The nontransformed cells were more susceptible to killing by MTX in Dulbecco's modified Eagle's medium (DMEM) than in MCDB 402, but the transformed cells were sensitive to MTX in both media. The high selectivity of MTX for transformed over nontransformed cells in MCDB 402 results from the presence of 1.0 microM leucovorin (5-formyltetrahydrofolate), a reduced form of the folic acid present in most other culture media. When leucovorin was added to DMEM with its high concentration of folic acid, the resistance to MTX of both nontransformed and transformed cells was greatly increased, but the selectivity of MTX for transformed cells was almost entirely lost. The results indicate that leucovorin protects nontransformed cells against concentrations of MTX that kill transformed cells, but the protection is dependent on the relative amounts of leucovorin to folic acid in the medium. The relative sensitivities of transformed and nontransformed cells in our system to MTX when both cell types are exhibiting their characteristic differential in growth behavior is similar to that described for tumor and normal cells in vivo. Since the unregulated growth behavior of the transformed, tumor-producing cells is efficiently and quantitatively measured in this system, it can be used to develop general principles of treatment and resolve questions of cytotoxic mechanism. PMID- 10462204 TI - A new human prostate carcinoma cell line, 22Rv1. AB - A cell line has been derived from a human prostatic carcinoma xenograft, CWR22R. This represents one of very few available cell lines representative of this disease. The cell line is derived from a xenograft that was serially propagated in mice after castration-induced regression and relapse of the parental, androgen dependent CWR22 xenograft. Flow cytometric and cytogenetic analysis showed that this cell line represents one hyper DNA-diploid stem line with two clonal, evolved cytogenetic sublines. The basic karyotype is close to that of the grandparent xenograft, CWR22, and is relatively simple with 50 chromosomes. In nude mice, the line forms tumors with morphology similar to that of the xenografts, and like the parental CWR22 and CWR22R xenografts, this cell line expresses prostate specific antigen. Growth is weakly stimulated by dihydroxytestosterone and lysates are immunoreactive with androgen receptor antibody by Western blot analysis. Growth is stimulated by epidermal growth factor but is not inhibited by transforming growth factor-beta1. PMID- 10462205 TI - The postnatal age of rat lung fibroblasts influences G1/S phase transition in vitro. AB - In the neonatal rat lung, alveolar development occurs from postnatal Days 4-13, during which time there is a fourfold increase in interstitial fibroblasts. Factors influencing emergence of new septa and cell proliferation associated with septal elongation have yet to be identified, in part because of difficulties inherent in studying this process in vivo. Using flow cytometric analysis of the DNA content of freshly isolated lung fibroblasts, we found that proliferation, as indicated by the percentage of cells in S plus G2/M phases, peaked on postnatal Day 4 (P < 0.04). By Days 9-10 the proliferation rate was lower than on Days 3, 4, 5, or 6 (P < 0.005). We then evaluated rates of in vitro proliferation as a function of postnatal age in first passage fibroblasts and found that the proliferative phenotype expressed in vivo persists in vitro. Fibroblasts from 4-5 d-old pups increased in number and incorporated 3H-thymidine at a faster rate than did fibroblasts obtained from pups at other postnatal ages (P < 0.0001). Age dependent differences in cell cycle transit time were compared in fibroblasts synchronized by serum starvation and analyzed by flow cytometry at 2-h intervals from 13-21 h after release from serum starvation. A greater percentage of cells from 5-d-old pups entered S phase during this period than was seen for cells obtained from 2-, 9-, 13-, or 23-d-old rat pups (P = 0.0001). Cells from 5-, 9-, and 13-d-old pups reentered G0/G1 by 21 h after release from serum starvation, in contrast to fibroblasts from 2- and 23-d-old rats which did not. Throughout the 15-h period after release from serum starvation, levels of cyclin E, which peaks at the G1/S border, were highest in the 5-d-old cells (P < 0.025). Synchronization with 2.5 mM hydroxyurea which inhibits DNA synthesis completely abolished age-related differences in cell cycle transit time, implying that age dependent differences in lung fibroblast proliferation rates are the result of events occurring before S-phase entry. PMID- 10462207 TI - Objective measurement of sagittal laxity of the knee. PMID- 10462206 TI - The effects of cell density, attachment substratum and dexamethasone on spontaneous apoptosis of rat hepatocytes in primary culture. AB - The rates of spontaneous cell detachment, cell viability, and apoptosis in primary cultures of rat hepatocytes plated at high and low density were compared. Apoptosis was frequent in detached cells, and the rates of cell detachment and apoptosis were greater in high-density than in low-density cultures. Among attached cells, more cells had condensed or fragmented nuclei in high-density than in low-density cultures. Further, ladder-like DNA fragmentation was not seen in low-cell-density cultures but was clearly evident in high-density cultures. Bax was more highly expressed in cells cultured at high density, and on collagen vs. matrigel, whereas changes of Bcl-2 and Fas expression observed in culture appeared unrelated to the rate of apoptosis. The rate of hepatocyte apoptosis appeared to be identical in low-density cultures on collagen 1 and matrigel, but when cells were cultured at high density, matrigel suppressed apoptosis by more than 50% at 36 h. In hepatocytes cultured on collagen 1, dexamethasone (0.1 microM) suppressed apoptosis in both low- and high-density cultures; higher doses had no further effects. In high density cultures, aurintricarboxylic acid (10 microM) suppressed apoptosis and this improved cell attachment at 48 h. It is concluded that cell viability in primary cultures of rat hepatocytes grown on collagen I is dependent on optimal culture density and that the cell population is regulated, at least in part, by apoptosis. Corticosteroids suppress spontaneous apoptosis of cultured hepatocytes in a non-dose-dependent manner, whereas matrigel abolishes apoptosis induced by increasing cell density. Bax may be an important protein in the cell density and cell matrix-dependent regulation of apoptosis in cultured hepatocytes. PMID- 10462208 TI - Evaluation of anterior knee joint instability with the Rolimeter. A test in comparison with manual assessment and measuring with the KT-1000 arthrometer. AB - We report on a comparative measurement of anterior knee joint laxity on 30 healthy test subjects and 30 patients with anterior knee joint instability using a new knee testing device (Rolimeter) and the KT- 1000 arthrometer compared to manual assessment of the anterior tibial translation in 30 degrees flexion (Lachman's test). We wanted to establish the reliability of the Rolimeter device compared to manual assessment and to the KT- 1000 arthrometer. The statistical evaluation by means of an analysis of variance (ANOVA) at a significance level of 5% showed no significant difference in the exactitude of measurement between the Rolimeter and the KT-1000 arthrometer, but altogether a significant difference between both instrumental methods compared with manual assessment. Thus the Rolimeter provides an economic, exact and simply operating device for quantifying anterior knee joint instability. PMID- 10462209 TI - Correlation of the intercondylar notch width of the femur to the width of the anterior and posterior cruciate ligaments. AB - The purpose of this study was to determine if a correlation exists between the intercondylar notch width (NW) of the femur and the width of the anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL). A study group of 124 consecutive patients (mean age 36.6 +/- 15.2 years; 67 men, 57 women) underwent a magnetic resonance imaging evaluation for knee pain but did not have an ACL or PCL tear or arthrosis. A T2 weighted coronal cut was identified and was located at the middle of the tibial spine, which represented the plane where the ACL and PCL cross each other when the knee is in 10 degrees of flexion. The NW and the width of the ACL and PCL were measured at the level of the middle of the popliteal hiatus on a physician-independent console that allowed for digital measurements in millimeters. Our results showed a statistically significant correlation between NW and ACL width (r = 0.87; P < 0.001) and between NW and PCL width (r = 0.75; P < 0.001). The mean ACL width was 6.4 +/- 1.4 mm (range 3-10 mm). The mean PCL width was 10.2 +/- 2.0 mm (range 6-17 mm). The mean ACL width was 5.7 +/- 1.1 mm for women and 7.1 +/- 1.2 mm for men (P < 0.001). The mean PCL width was 9.5 +/- 1.7 mm for women and 10.9 +/- 2.0 for men (P < 0.001). Our results indicate that NW correlates with ACL and PCL width. In addition, ACL and PCL widths are narrower in women than men. PMID- 10462210 TI - Hamstring graft motion in the femoral bone tunnel when using titanium button/polyester tape fixation. AB - The objective of this study was to determine the relative motion of a quadruple hamstring graft within the femoral bone tunnel (graft-tunnel motion) under tensile loading. Six graft constructs were prepared from the semitendinosus and gracilis tendons of human cadavers and were fixed with a titanium button and polyester tape within a bone tunnel in a cadaveric femur. Three different lengths of polyester tape (15, 25, and 35 mm loops) were evaluated. The femur was held stationary and uniaxial tensile loads were applied to the distal end of the graft using a materials testing machine. Each construct was subjected to loading for ten cycles with upper limits of 50 N, 100 N, 200 N and 300 N. Graft-tunnel motion was then determined using the distances between reflective tape markers placed on the hamstring graft and at the entrance to the femoral bone tunnel, which were tracked with a high-resolution video system. Graft-tunnel motion was found to range from 0.7 +/- 0.2 mm to 3.3 +/- 0.2 mm, and significant increases in graft tunnel motion were observed with increasing tensile loads (P < 0.05). Shorter tape length (15 mm) resulted in significantly less motion when compared to longer tape length (35 mm) (P < 0.05). We conclude that graft-tunnel motion is significant and should be considered when using this fixation technique. Early stress on the graft, as seen in postoperative rehabilitation exercises and athletic activities, may cause large graft-tunnel motion before graft incorporation is complete. A shorter distance between the tendon tissue and the titanium button is recommended to minimize the amount of graft-tunnel motion. Alternative fixation materials to polyester tape, or different fixation techniques, need to be developed such that graft-tunnel motion can be reduced. Further studies are needed to evaluate the effect of graft-tunnel motion on graft incorporation in the bone tunnel. PMID- 10462211 TI - Semitendinosus tendon regeneration after harvesting for ACL reconstruction. A prospective MRI study. AB - Utilisation of the semitendinosus and gracilis tendons in reconstruction of the anterior cruciate ligament (ACL) has become more common during the last few years. In recent studies a regeneration potential in the harvested tendons has been observed. In this study, 11 consecutive patients who underwent ACL reconstruction with a quadruple semitendinosus graft were examined 6-12 months postoperatively by MRI. Another two patients were examined within 2 weeks after surgery. The median age of the patients was 24 years and there were 8 males and 3 females. The right knee was involved in six patients and the left knee in five. A low-field 0.2 Tesla Siemens open MRI was used for examinations and T1 and T2 weighted transaxial sequences over the thigh and the knee joint were performed. In some instances, additional sagittal sequences were used. ROI analysis of the pixel value of the signal and area determinations on transaxial sequences was performed for both the involved and the healthy side. In 8 of the 11 patients examined 6-12 months postoperatively, a regeneration of the semitendinosus tendon with normal anatomical topographies to the level of the tibial plateau was found. Three of these eight patients were analysed more distally and fusion of the semitendinosus and gracilis tendons was found approximately 30 mm below the joint line before they inserted as a "conjoined tendon" into the pes anserinus. At the mid-thigh level, the semitendinosus muscle had a smaller area and a higher signal than that on the normal side. However, this difference was smaller in the patients showing normal distal tendon regeneration. This study indicates that the semitendinosus tendon has a strong potential for regeneration and that the muscle atrophy seems to be less in the patients with a more normalised distal insertion of the tendon in the pes anserinus. PMID- 10462212 TI - Proprioceptive defects after an anterior cruciate ligament rupture -- the relation to associated anatomical lesions and subjective knee function. AB - A disturbed proprioception has been described in patients with an anterior cruciate ligament (ACL) deficient knee. The relation to demographic data and to different commonly associated anatomical lesions, as well as to subjective knee function, was prospectively studied in 16 consecutive patients after an acute knee ligament injury. All patients had a complete rupture of the ACL, but variable associated anatomical lesions. The threshold to detect a passive motion, as a measure of their proprioceptive ability, was registered repeatedly during the first year after injury. Four of the patients had consistently severe and persistent deficits at 1, 2 and 8 months. These four individuals had more chondral lesions and a lower subjective rating of their knee function than the remaining patients. In the whole group there were significant correlations between the recorded thresholds and associated chondral lesions, meniscal lesions and the subjective rating of knee function. We found no significant relation between age, gender, activity level, grade of mechanical laxity increase or a medial collateral ligament rupture, and the proprioceptive recordings. Thus, morphological lesions other than a rupture of the ACL seem to contribute to the proprioceptive deficits after a knee ligament injury, and the patients' ability to detect a passive motion showed a relation to subjective knee function from the time of injury onwards. PMID- 10462213 TI - Superior results with continuous passive motion compared to active motion after periosteal transplantation. A retrospective study of human patella cartilage defect treatment. AB - Fifty-seven consecutive patients (33 men and 24 women), with a mean age of 32 years (range 16-53 years), who suffered from an isolated full-thickness cartilage defect of the patella and disabling knee pain of long duration, were treated by autologous periosteal transplantation to the cartilage defect. The first 38 consecutive patients (group A) were postoperatively treated with continuous passive motion (CPM), and the next 19 consecutive patients (group B) were treated with active motion for the first 5 days postoperatively. In both groups, the initial regimens were followed by active motion, slowly progressive strength training, and slowly progressive weight bearing. In group A, after a mean follow up of 51 months (range 33-92 months), 29 patients (76%) were graded as excellent or good, 7 patients (19%) were graded as fair, and 2 patients (5%) were graded as poor. In group B, after a mean follow-up of 21 months (range 14-28 months), 10 patients (53%) were graded as excellent or good, 6 patients (32%) were graded as fair, and 3 patients (15%) were graded as poor. Altogether, nine of the fair or poor cases (50%) were diagnosed with chondromalacia of the patella. Our results, after performing autologous periosteal transplantation in patients with full thickness cartilage defects of the patella and disabling knee pain, are good if CPM is used postoperatively. The clinical results using active motion postoperatively are not acceptable, especially not in patients with chondromalacia of the patella. PMID- 10462214 TI - Jumper's knee: postoperative assessment. A retrospective clinical study. AB - Jumper's knee or infrapatellar insertional tendinopathy is a condition primarily found in athletes between 18 and 25 years of age who are engaged in explosive running and jumping sports. It is caused by microtears or partial macrotears through the patellar tendon. Conservative treatment is used as the primary approach. If conservative measures are insufficient or fail to relieve the symptoms, surgery is indicated. The patellar tendon is incised and the hyaline inflammatory tissue is removed. Twenty-six patients have been studied retrospectively, with specific attention to postoperative resumption of sports and residual subjective and objective findings. They all completed a detailed questionnaire and subsequently underwent a physical and ultrasound examination. PMID- 10462215 TI - Fractures of the patella in children. AB - Fractures of the patella are relatively rare injuries for children. Fourteen patients with patellar fractures were reviewed. There were 12 boys and two girls with the age ranging between 9 and 15 years and an average of 11.4 years. Sleeve fractures were the most common type of patellar fractures observed (eight cases), followed by transverse fractures (four cases) and comminuted fractures (two cases). All the fractures were treated with open reduction and followed up for 2 to 20 years. Overall results were good in 13 patients, and flexion limitation of the knee was seen in one. Fractures of the patella in children are characterised by sleeve fractures. The prognosis of the patients treated with open reduction is generally good. PMID- 10462216 TI - Arterial injury during revision total knee replacement. A case report. AB - A case of an anatomical variation of the arterial vessels in the popliteal fossa, leading to a vascular transection during a total knee revision is presented. The authors believe this complication should be considered in all cases of knee revision surgery. PMID- 10462217 TI - Pigmented villonodular synovitis of the shoulder: review and case report. AB - Pigmented villonodular synovitis (PVNS) as reviewed in detail elsewhere most frequently involves the knee and finger synovial structures; shoulder involvement is rare: A search through the English literature yielded 18 publications describing 25 cases of PVNS affecting the shoulder joint. Analyzing these reports we found the clinical and radiological findings generally to be nonspecific, often mimicking a malignancy, as in the case presented here of a 16-year-old boy with painful swelling in the area of the left proximal humerus. Magnetic resonance imaging showed a suspected malignant soft tissue mass involving the shoulder capsule and measuring 7.5 x 6 x 4 cm. Preoperatively the patient could recall no trauma; however, postoperatively he did report a distortion trauma of the affected shoulder following a bicycle accident. Intraoperatively, two tumors were found infiltrating the axillary vessels and nerve and tendon structures originating in the capsule of the shoulder joint. Rapid sections of the tissue revealed no signs of malignancy; further pathohistological examination revealed localized PVNS. Preoperatively, the shoulder joint was not suspected as the primary site of origin of the tumor because the patient had no complaints or functional deficits of the shoulder. The clinical presentation of such a PVNS lesion over the proximal humerus is unusual and to date has only twice been described in the literature. PMID- 10462218 TI - Hamstring extensibility and transverse plane knee control relationship in athletic women. AB - Athletic women are at particular risk for sustaining a non-contact anterior cruciate ligament (ACL) injury. The hamstrings are vital to providing dynamic knee motion control in the sagittal and transverse planes during running stance, and some have suggested this function is enhanced when they are less extensible. This study attempted to determine the correlational relationships between hamstring extensibility and transverse plane knee kinematics and from these data to describe the dynamic transverse plane knee motion control capabilities of the hamstrings. Twenty normal athletic women (mean +/- SD; aged 21 +/- 1.6 years; height 163.3 +/- 5.7 cm; weight 60.1 +/- 3.6 kg) were evaluated for active hamstring extensibility and transverse plane knee kinematic relationships during crossover-cut stance phase. Following crossover-cut training (3 weeks) using the left (preferred) lower extremity as the stance limb, hamstring extensibility was measured. Following this, subjects were fitted with 9 retroreflective markers denoting the local segmental coordinate systems (3 markers each) of the left foot, leg and thigh. Kinematic (3-dimensional, four phase-locked cameras, 200 Hz) knee and ankle data were sampled and analyzed. Descriptive statistics and Pearson correlations between hamstring extensibility and knee and ankle kinematic variables were calculated. Statistical significance was set at P < or = 0.05 with Bonferroni adjustments. Hamstring extensibility (12 degrees +/- 8 degrees terminal extension) revealed low but significant positive correlations with tibial external rotation (6 degrees +/- 10.7 degrees) at heelstrike (19.3 degrees +/- 8.5 degrees knee flexion) (r = 0.62, P = 0.004) and tibial internal rotation (-13 degrees +/- 8.4 degrees) at peak knee flexion (57.8 degrees +/- 9.3 degrees) following heelstrike (r = 0.47, P = 0.01). Increased hamstring extensibility resulted in increased tibial external rotation at heelstrike and decreased tibial internal rotation at peak knee flexion. Increased hamstring extensibility may improve knee extensor efficiency at heelstrike by enabling greater tibial external rotation and protect the ACL at peak knee flexion by decreasing the tibial internal rotation magnitude. PMID- 10462219 TI - Tibia and fibula fractures in soccer players. AB - We performed a retrospective review of 31 athletes who sustained a fracture of the lower leg from a direct blow while playing soccer. Fifteen fractures involved both the tibia and fibula 11 only the tibia, and 5 only the fibula. Information was collected using a standardized questionnaire. The mean follow-up from the time of injury was 30 months. Injuries typically occurred in young, competitive athletes during game situations. The mechanisms were broadly classified into several categories: contact during a slide tackle (13, 42%), a collision with the goalkeeper (8, 26%), two opposing players colliding while swinging for a loose ball (7, 23%), or a player being kicked by a standing opponent (3, 10%). The majority of fractures (26, 90%) occurred while the athletes were wearing shin guards. The point of impact was with the shin guard prior to the fracture in 16 cases (62%). Return to competitive soccer averaged 40 weeks for combined tibia and fibula fractures, 35 weeks for isolated tibia fractures, and 18 weeks for isolated fibula fractures. Injuries were associated with a high incidence of major complications (12 out of 31, 39%), especially in concurrent tibia and fibula fractures (8 out of 15, 50%). These findings suggest that lower leg fractures in soccer players are serious injuries, often necessitating a prolonged recovery time. In addition, this study questions the ability of shin guards to protect against fractures. PMID- 10462220 TI - Increase of metallothionein-immunopositive chloride cells in the gills of brown trout and rainbow trout after exposure to sewage treatment plant effluents. AB - Metallothionein, a biomarker of exposure and toxicity of heavy metals, has been detected in the gills of brown trout (Salmo trutta fario L.) and rainbow trout (Oncorhynchus mykiss Richardson) by means of immunohistochemistry. A very prominent labelling of chloride cells was found after exposure to diluted sewage plant effluents. No significant increase was observed in either the number of labelled cells or their labelling intensity after exposure to water of a polluted river compared to fish kept in tap water. These results do not correlate with findings of a histopathological study, suggesting that the metal levels at the sewage treatment plant were too low to produce gross histopathology. A comparison between the species indicated that the rainbow trout showed a generally higher metallothionein expression than the brown trout. PMID- 10462221 TI - Cytochemical localization of calcium in prefusion myoblasts from the chick embryo myotome. AB - Myoblast fusion is a Ca2+-dependent process. The aim of this report was to study the localization of Ca2+ in prefusion myoblasts from the brachial somites of chick embryos (51-108 h of incubation), using the potassium pyroantimonate cytochemical method. When observed under a transmission electron microscope, electron-dense precipitates of Ca2+-antimonate were found in the basement membrane of the myotome, which separates the myotome from the adjacent mesenchyma. Within myoblasts, triads and sarcoplasmic reticulum associated with the first newly formed sarcomeres were observed, but a T-tubule network was not found. Moreover, Ca2+-antimonate precipitates were not observed in structures resembling T-tubules or sarcoplasmic reticulum. The results suggest that sarcomerogenesis and sarcoplasmic reticulum development occur simultaneously and that prefusion myoblasts have neither a T-tubule network nor Ca2+ deposits on sarcoplasmic reticulum. Small Ca2+ pools were found in the myoblast nuclei, cytoplasmic vesicles and mitochondrias. Ca2+-antimonate precipitates periodically distributed at the cell periphery, close to the cell membrane, were observed. These precipitates could represent internal Ca2+ stores located in the peripheral couplings and it is proposed that these pools of Ca2+ could be mobilized before fusion, leading to the increase in free intracellular Ca2+ that precedes myoblast fusion. PMID- 10462222 TI - Oxidative myocytes of heart and skeletal muscle express abundant sarcomeric mitochondrial creatine kinase. AB - Sarcomeric mitochondrial creatine kinase catalyzes the reversible transfer of a high energy phosphate between ATP and creatine. To study cellular distribution of the kinase, we performed immunocytochemical studies using a peptide antiserum specific for the kinase protein. Our results demonstrated that the sarcomeric mitochondrial creatine kinase gene is abundantly expressed in heart and skeletal muscle, with no protein detected in other tissues examined, including brain, lung, liver, spleen, kidney, bladder, testis, stomach, intestine, and colon. RNA blot study showed that there is no detectable expression of the kinase mRNA in the thymus gland. In heart and skeletal muscle, the kinase protein is expressed in atrial and ventricular cardiomyocytes and a subpopulation of skeletal myofibres. In skeletal muscle, fast myosin heavy chain co-localization studies demonstrated that the sarcomeric mitochondrial creatine kinase is highly expressed in type 1, slow-oxidative and type 2A, fast-oxidative-glycolytic myofibres. We conclude that the kinase gene is abundantly expressed in oxidative myocytes of heart and skeletal muscle and may contribute to oxidative capacity of these cells. PMID- 10462223 TI - The distribution of BrdU- and TUNEL-positive cells during odontogenesis in mouse lower first molars. AB - This study investigated the minute distribution of both proliferating and non proliferating cells, and cell death in the developing mouse lower first molars using 5-bromo-2'-deoxyuridine (BrdU) incorporation and the terminal deoxynucleotidyl transferase-mediated deoxyuridine-5'-triphosphate (dUTP)-biotin nick end labeling (TUNEL) double-staining technique. The distribution pattern of the TUNEL-positive cells was more notable than that of the BrdU-positive cells. TUNEL-positive cells were localized in the following six sites: (1) in the most superficial layer of the dental epithelium during the initiation stage, (2) in the dental lamina throughout the period during which tooth germs grow after bud formation, (3) in the dental epithelium in the most anterior part of the antero posterior axis of the tooth germ after bud formation, (4) in the primary enamel knot from the late bud stage to the late cap stage, (5) in the secondary enamel knots from the late cap stage to the late bell stage, and (6) in the stellate reticulum around the tips of the prospective cusps after the early bell stage. These peculiar distributions of TUNEL-positive cells seemed to have some effect on either the determination of the exact position of the tooth germ in the mandible or on the complicated morphogenesis of the cusps. The distribution of BrdU-negative cells was closely associated with TUNEL-positive cells, which thus suggested cell arrest and the cell death to be essential for the tooth morphogenesis. PMID- 10462224 TI - Identification of cell types in the developing goat mammary gland. AB - The goat was chosen as the model system for investigating mammary gland development in the ruminant. Histological and immunocytochemical staining of goat mammary tissue at key stages of development was performed to characterize the histogenesis of the ruminant mammary gland. The mammary gland of the virgin adult goat consisted of a ductal system terminating in lobules of ductules. Lobuloalveolar development of ductules occurred during pregnancy and lactation which was followed by the regression of secretory alveoli at involution. The ductal system was separated from the surrounding stroma by a basement membrane which was defined by antisera raised against laminin and Type IV collagen. Vimentin, smooth-muscle actin and myosin monoclonal antisera as well as antisera to cytokeratin 18 and multiple cytokeratins stained a layer of myoepithelial cells which surround the ductal epithelium. Staining of luminal epithelial cells by monoclonal antibodies to cytokeratins was dependent on their location along the ductal system, from intense staining in ducts to variable staining in ductules. The staining of epithelial cells by monoclonals to cytokeratins also varied according to the developmental status of the goat, being maximal in virgin and involuting glands, lowest at lactation and intermediate during gestation. In addition, cuboidal cells, situated perpendicular to myoepithelial cells and adjacent to alveolar cells in secretory alveoli, were also stained by cytokeratin monoclonal antibodies and antisera to the receptor protein, erbB-2, in similar fashion to luminal epithelial cells. These results demonstrate that caprine mammary epithelial cell differentiation along the alveolar pathway is associated with the loss of certain types of cytokeratins and that undifferentiated and secretory alveolar epithelial cells are present within lactating goat mammary alveoli. PMID- 10462225 TI - Two peptidergic drugs increase the synaptophysin immunoreactivity in brains of 24 month-old rats. AB - The brain-derived peptidergic drug Cerebrolysin has been found to support the survival of neurones in vitro and in vivo. Positive effects on learning and memory have been demonstrated in various animal models and also in clinical trails. In the present study, the effects of Cerebrolysin and its peptide preparation E021 on the synapse density in the hippocampus, the dentate gyrus and in the entorhinal cortex of 24-month-old rats were investigated. Rats received the drugs or saline for control for 19 consecutive days (2.5 ml/kg per day). Slices of the brains were immunohistochemically stained with anti-synaptophysin, which is a specific marker of presynaptic terminals. Quantification of the synapse density was done by using light microscopy and a computerised image analysing system. Our results clearly showed that the rats benefit from the administration of both drugs, showing an enhancement in the number of synaptophysin-immunostained presynaptic terminals in the entorhinal cortex, the dentate gyrus, and also in the hippocampal subfields CA1, CA2, CA3 stratum lucidum and CA3 stratum radiatum. It can be assumed that these effects are the reason for improved cognitive performances of rats treated with Cerebrolysin and E021. PMID- 10462226 TI - Effects of bone morphogenetic protein-2 and transforming growth factor-beta1 on gene expression of decorin and biglycan by cultured osteoblastic cells. AB - The influence of bone morphogenetic protein-2 (BMP-2) and transforming growth factor beta (TGF-beta) on the expression of small proteoglycans, decorin and biglycan was investigated in a clonal rat osteoblastic cell line, ROS-C26 (C26) cells, which is a potential osteoblast precursor cell line and capable of differentiating into mature osteoblasts after treatment with recombinant BMP-2 (rhBMP-2). Following the culture of C26 cells for 3, 6, and 9 days in the presence or absence of rhBMP-2, alkaline phosphatase activity increased in the rhBMP-2 treated cells in direct proportion to their differentiation into more mature osteoblastic cells, whereas decorin mRNA decreased in the cells, when compared to control cells without rhBMP-2 treatment. These results were evident 6 days after treatment. However, rhBMP-2 treatment had no effect on biglycan mRNA expression in the cells. Subsequently, after removal of rhBMP-2 from the culture media, the cells were further cultured for 24 h with graded concentrations of TGF beta1 (0, 0.1, 1.0, 5.0, and 10 ng/ml). TGF-beta1 decreased decorin mRNA expression in the cells dose dependently, but did not affect their biglycan mRNA expression. Furthermore, either removal of rhBMP-2 from the culture media or addition of TGF-beta1 significantly decreased alkaline phosphatase activity of rhBMP-2-induced cells. These results indicate that osteoblastic differentiation is accompanied by increased alkaline phosphatase activity and decreased expression of decorin mRNA, but continuous expression of biglycan mRNA. Both rhBMP-2 and TGF-beta1 inhibit decorin mRNA expression in osteoblasts at varying stages of differentiation, but their effects on biglycan mRNA expression and alkaline phosphatase are different. PMID- 10462227 TI - Impaired antibody response after immunization of HIV-infected individuals with the polysaccharide vaccine against Salmonella typhi (Typhim-Vi). AB - Infections with Salmonella species, including Salmonella typhi, are more frequently observed in HIV-infected individuals than in healthy individuals. HIV infected individuals were vaccinated with polysaccharide vaccine against Salmonella typhi (Typhim-Vi) which is assumed to be a T-cell-independent antigen. We found that the antibody response in patients with < 200 x 10(6)/l CD4+ T lymphocytes was significantly lower compared with patients with > or = 200 x 10(6)/l CD4+ T lymphocytes and healthy controls. The antibody response after vaccination with the polysaccharide salmonella Vi-antigen was correlated with the number of CD4+ T lymphocytes and therefore Typhim-Vi can be considered to be a T cell-independent type 2 antigen. The results of this study indicate that after vaccination the proportion of HIV-infected individuals with protective antibody concentrations against Salmonella typhi will be lower than in healthy controls. PMID- 10462228 TI - Hepatitis B vaccination in infancy in The Gambia: protection against carriage at 9 years of age. AB - To estimate the efficacy in The Gambia (West Africa) of infant hepatitis B vaccination against infection and carriage with the virus at the age of 9 years. The HBV status of 9-year old children vaccinated in infancy was compared to that of unvaccinated children of the same age. Eight percent of the vaccinated children had been infected by HBV compared to 50% of the unvaccinated control group; HBV carrier status was 0.6 and 10% respectively, resulting in a vaccine efficacy of 83% against infection and of 95% against chronic carriage. The results show that infant vaccination provides a high level of protection at the age of nine years against both HBV infection and chronic carrier status and no booster dose of vaccine is required in the first decade. These findings support the WHO recommendation for the introduction of HBV vaccination into the Expanded Programme on Immunization in Africa. PMID- 10462229 TI - Neisseria meningitidis serogroup C polysaccharide and serogroup B outer membrane vesicle conjugate as a bivalent meningococcus vaccine candidate. AB - Neisseria meningitidis serogroup C polysaccharide (PS C) was conjugated to serogroup B outer membrane vesicles (OMV) in order to test the possibility of obtaining a bivalent group B and C meningococcus vaccine. The conjugate and controls were injected intraperitoneally into groups of ten mice with boosters on days 14 and 28 after the primary immunization. The following groups were used as control: (i) PS C; (ii) PS C plus OMV; (iii) OMV; and (iv) saline. The serum collected on days 0, 14, 28 and 42 were tested by enzyme-linked immunosorbent assay (ELISA) for PS C and OMV, and by complement mediated bactericidal assay against serogroups B and C. ELISA for PS C as well as bactericidal titres against serogroup C meningococci of the conjugated vaccine increased eight-fold (ELISA) and 32 fold (bactericidal) after 42 days in comparison with the PS C control group. ELISA for OMV and bactericidal titre against serogroup B meningococci of the conjugate showed no significant difference in comparison with the OMV containing controls. Furthermore, Western Blot assay of the conjugate immune serum did not bind OMV class four protein which is related to the complement dependent antibody suppressor. The results indicate that the PS C-OMV conjugate could be a candidate for a bivalent vaccine toward serogroups B and C meningococci. PMID- 10462230 TI - Antigenic properties of the merozoite surface protein 1 gene of Plasmodium vivax. AB - Plasmodium vivax is responsible for an approximate 35 million yearly human cases of malaria. Unfortunately, due to the low mortality rate associated with it and the difficulties of continuously in vitro culturing of this parasite, vaccine development against this human malaria has been largely neglected. In here, the antigenic properties of the merozoite surface protein 1 gene of P. vivax (PvMSP 1), were studied. Thus, seven recombinant bacterial plasmids coding different regions of the PvMSP-1 protein were constructed and used to immunize BALB/c mice. The results demonstrated that a plasmid encoding the entire N-terminus comprising 682 amino acids and a plasmid encoding the C-terminus including the two juxtaposed epidermal growth factor (EGF)-like domains fused to the Hepatitis B surface antigen, were antigenic. Moreover, the elicited immune responses were similar to those reported for these same PvMSP-1 regions in natural human infections. PMID- 10462231 TI - Potency test of inactivated Newcastle disease vaccines by monoclonal antibody blocking ELISA. AB - The potency of 27 different inactivated Newcastle disease (ND) vaccines was determined by immunization and challenge tests. Three- and four-week-old SPF chickens were immunized with 1/50 dose of vaccines. The chickens were bled and challenged three weeks later. The protected/challenged ratio was determined. Serum samples were tested by the haemagglutination inhibition (HI) test and by a monoclonal antibody (MAB) blocking enzyme-linked immunosorbent assay (B-ELISA). Ninety-four percent of the vaccinated chickens (353 birds) with a B-ELISA value above 60% were protected. A strong positive correlation (0.934) was found between 'actual protection' (protection against challenge) and 'estimated protection' (protection calculated from the B-ELISA results). Based upon the results obtained, this B-ELISA seems to be suitable for replacing the challenge test in the potency control of inactivated ND vaccines in the future. PMID- 10462232 TI - Generation of antigen specific CD8+ cytotoxic T cells following immunization with soluble protein formulated with novel glycoside adjuvants. AB - Presentation of peptide on MHC class I molecules is essential to elicit cytolytic T cell (CTL) activity. Such peptides are a result of the cytosolic, or class I, antigen processing pathway. Due to the segregation of the class I and the exogenous processing pathway, soluble protein cannot enter the class I pathway and is thus incapable of inducing CTL. However careful formulation with adjuvants can overcome this obstacle. In this study we evaluated the capacity of two novel amphiphilic adjuvants, better termed delivery vehicles, to elicit CTL activity in a C57Bl/6 murine model with ovalbumin (OVA) as an antigen. Incomplete Freund's adjuvant and aluminium hydroxide (Alhydrogel) were used as reference adjuvants. In addition the oil-in-water emulsion Provax was used throughout as a positive control adjuvant. Both amphiphile preparations were capable of eliciting potent CTL activity after administration of one immunizing dose of ovalbumin. CTL were CD8+ restricted as assessed by in vitro depletion of CD8+ and CD4+ T cells. CTL activity was also MHC-restricted as well as specific for the H-2Kb OVA motif SIINFEKL. PMID- 10462233 TI - Dose-dependent effects of IL-12 treatment to immune response induced after immunization with a recombinant respiratory syncytial virus (RSV) fusion protein fragment. AB - The amino acid (aa) sequence 190-289 of the RSV fusion (F) glycoprotein expressed in insect cells (bF(190-289)) has been shown to partially protect BALB/c mice and to prime for a Th2 cell response. We evaluated the effects of IL-12 treatment during antigen priming of bF(190-289) on immune response and protective efficacy. Low doses of IL-12 (10 ng) reduced IL-4 and IL-5 secretion (but did not affect IL 10 production) and decreased inflammatory signs whereas high doses of IL-12 had no effects. In addition, IL-12 treatment did not improve resistance to RSV replication. These results suggest that IL-12 treatment attenuates Th2 response and Th2 associated pulmonary inflammatory response in a dose-dependent manner, without improving protective efficacy. PMID- 10462234 TI - Cytotoxic T lymphocytes to Ebola Zaire virus are induced in mice by immunization with liposomes containing lipid A. AB - An eight amino acid sequence (TELRTFSI) present in the carboxy terminal end (aa 577-584) of membrane-anchored GP, the major structural protein of Ebola virus, was identified as an H-2k-specific murine cytotoxic T cell epitope. Cytotoxic T lymphocytes (CTLs) to this epitope were induced by immunizing B10.BR mice intravenously with either irradiated Ebola virus or with irradiated Ebola virus encapsulated in liposomes containing lipid A. The CTL response induced by irradiated Ebola virus could not be sustained after the second round of in vitro stimulation of immune splenocytes with the peptide, unless the irradiated virus was encapsulated in liposomes containing lipid A. The identification of an Ebola GP-specific CTL epitope and the requirement of liposomal lipid A for CTL memory recall responses could prove to be a promising approach for developing a vaccine against Ebola virus infection. PMID- 10462235 TI - An acellular pertussis vaccine in healthy adults: safety and immunogenicity. Pennridge Pediatric Associates. AB - Recent data indicate that Bordetella pertussis can be an important cause of illness in adolescents and adults. In a randomized observer- and subject-blinded study, adults (> or = 18 years of age) received an acellular pertussis (aP) vaccine containing genetically inactivated pertussis toxin (PT), filamentous hemagglutinin (FHA) and pertactin (PRN), or a saline placebo, and were monitored for safety and immunogenicity. IgG antibodies to PT, FHA, and PRN were measured by enzyme-linked immunosorbent assay (ELISA) and PT neutralization by a Chinese hamster ovary (CHO) cell assay. Local reactions, more common in the aP group, were mild and transient. One month after immunization, geometric mean ELISA antibody concentrations for the aP and placebo groups, respectively, were: anti PT, 463 and 7.6; anti-FHA, 417 and 18; and anti-PRN, 855 and 14. The anti-PT neutralization titers for the aP and placebo groups were 1:3439 and 1:58 respectively. This aP vaccine is a safe and immunogenic candidate booster vaccine against pertussis for adults. PMID- 10462236 TI - Immunization of mice with peptomers covalently coupled to aluminum oxide nanoparticles. AB - Subunit vaccines generally require adjuvants to elicit immune responses, but adjuvants may alter the conformation of critical epitopes and reduce vaccine efficacy. We therefore tested an immunization strategy in which antigen is covalently coupled to aluminum oxide nanoparticles using a method that favors preservation of the native conformation. The test antigen consisted of "peptomers" (head-to-tail-linked peptide homopolymers) derived from the 4th conserved region (C4) of HIV-1 gp120 which is believed to be in an alpha-helical conformation prior to binding to CD4. Immune responses in mice to peptomer nanoparticle conjugates were compared to responses elicited by free C4 peptide and C4 peptomers, with and without the hydrophilic adjuvant muramyl dipeptide (MDP). Highest peptomer-specific serum antibody responses were induced by peptomer-particles without MDP. Serum antibodies induced by peptomer-particles also showed highest reactivity towards recombinant, glycosylated gp120 and HIV-1 infected T cells. The results suggest that this novel vaccine approach could be useful for induction of immune responses against conformation-sensitive viral antigens without the need for additional adjuvants. PMID- 10462237 TI - Development of biopharmaceuticals in plant expression systems: cloning, expression and immunological reactivity of human cytomegalovirus glycoprotein B (UL55) in seeds of transgenic tobacco. AB - Plant seeds offer unique opportunities for the production and delivery of oral subunit vaccines. We have used the immunodominant glycoprotein B complex of human cytomegalovirus (HCMV), introduced into tobacco plants, as a model system for studying the merit of this promising approach. Given the advantages of expressing proteins in seeds, a novel expression vector was developed incorporating regulatory sequences of glutelin, the major rice seed storage protein, to direct synthesis of recombinant glycoprotein B. Analysis of genomic DNA of 28 selected tobacco transformants by PCR amplification showed that 71% harboured the gB cDNA, a finding further documented by Southern blotting. Specific immunoassays of protein extracts from seeds of positive plants showed that all were producing antigenic glycoprotein B at levels ranging from 70-146 ng/mg extracted protein. In addition, similarity with native glycoprotein B produced in HCMV-infected cells was also demonstrated by inhibition of immunofluorescence on HCMV-infected human fibroblasts. These data are the first to report the expression of an immunodominant antigen of HCMV in plant tissues, indicating the fidelity with which this very large heterologous viral glycoprotein can be synthesized in this model system. PMID- 10462238 TI - Manipulation of pathogen-derived genes to influence antigen presentation via DNA vaccines. AB - To gain insight into the routes of presentation of pathogen sequences via DNA vaccines, we have compared the abilities of sequences encoding fragment C of tetanus toxin (FrC) and influenza A virus nucleoprotein (NP) to induce antibody or cytotoxic T-cell (CTL) responses in vivo. Strong antibody and CTL responses were induced against FrC targeted to the endoplasmic reticulum (ER) and both were reduced by removal of the leader sequence. In contrast, targeting of NP to the ER generated only a modest antibody response, likely due to misfolding in this site. Removal of the leader sequence led to anti-NP antibodies via cross-priming. For NP, induction of CTLs was not influenced by the leader sequence. Exogenous FrC or NP delivered as proteins were unable to induce CTLs. Routes to induction of optimal immune responses via DNA evidently differ according to the nature of the encoded pathogen sequence. Understanding processing pathways for pathogen sequences should assist rational design of DNA vaccines. PMID- 10462239 TI - Differentiation of convalescent animals from those vaccinated against foot-and mouth disease by a peptide ELISA. AB - We have identified continuous antigenic determinants within the amino acid sequences of the conserved nonstructural region containing proteins 2C and 3ABC of foot-and-mouth disease virus which can distinguish between the sera from vaccinated and infected animals. An ELISA based on a 3B peptide gave a positive reaction with sera from cattle, pigs, sheep and guinea pigs infected with all seven serotypes of the virus, but not with sera from vaccinated animals. In experiments with cattle and pigs to determine the duration of the antibody response, positive reactions were obtained as late as one year after infection. The advantages of using peptides from the nonstructural viral proteins instead of recombinant proteins for differentiating vaccinees from infected animals include their exquisite specificity, nonreactivity with antibodies against host cell derived proteins (e.g. E. coli and insect cell proteins), and their ease of preparation. PMID- 10462240 TI - The induction of systemic and mucosal immune responses following the subcutaneous immunization of mature adult mice: characterization of the antibodies in mucosal secretions of animals immunized with antigen formulations containing a vitamin D3 adjuvant. AB - Systemic and mucosal immune responses were effectively induced following the subcutaneous administration of Haemophilus influenzae type b oligosaccharide conjugated to diphtheria toxoid vaccine in a formulation containing the active form of vitamin D3. IgA and IgG antibodies with specificity for both the protein and oligosaccharide components of the vaccine were detectable in mucosal secretions following immunization. The IgA and IgG mucosal antibodies were produced locally, and were functional as demonstrated by their diphtheria toxin neutralizing activity. Our data suggests that subcutaneous tissues can effectively serve as effective antigen presenting sites for both mucosal and systemic immune responses to antigens administered in combination with vitamin D3. PMID- 10462241 TI - Synthetic oligodeoxynucleotides containing CpG motifs enhance immunogenicity of a peptide malaria vaccine in Aotus monkeys. AB - Synthetic peptide and recombinant protein vaccines are optimally immunogenic when delivered with an effective adjuvant. Candidate vaccines currently insufficiently immunogenic may induce a protective immunity if they could be delivered with more effective adjuvants. For example, immunogens that induce promising responses when administered to mice with complete and incomplete Freund's adjuvants perform less well in primate animal models where complete Freund's adjuvant is not used. We report the use of synthetic oligodeoxynucleotides containing CpG motifs, the sequences of which are based on immunostimulatory bacterial DNA sequences, to enhance the immune response in Aotus monkeys to a synthetic peptide malaria vaccine. Monkeys were immunized with the synthetic peptide PADRE 45, a synthetic peptide containing amino acid sequences derived from the circumsporozoite protein (CSP) from Plasmodium falciparum, and delivered in an emulsion of saline and Montanide 720, a mannide oleate in oil solution, that also contained one of three oligodeoxynucleotides. The animals receiving oligodeoxynucleotides containing either three or four CpG motifs produced antibodies that bound a recombinant CSP as measured in ELISA, and reacted with P. falciparum sporozoites in a sporozoite immunofluorescent test. These responses were significantly greater than those seen in animals receiving the oligodeoxynucleotide without CpG motifs. These data indicate that oligodeoxynucleotides containing CpG motifs improve immunogenicity of peptide immunogens in non-human primates, and may be immunopotentiators useful in humans. PMID- 10462242 TI - Immunization of rhesus monkeys with a mucosal prime, parenteral boost strategy protects against infection with Helicobacter pylori. AB - Rhesus monkeys were immunized with recombinant Helicobacter pylori urease vaccine given solely by the parenteral route or preceded by a priming dose given by the oral route. Two groups of monkeys received parenteral urease with either a synthetic glycolipid adjuvant (Bay) or aluminum hydroxide (alum) as adjuvants. A third group of monkeys received a priming dose of oral urease given with the mucosal adjuvant LT (Escherichia coli heat labile enterotoxin), followed by parenterally administered booster doses of urease adsorbed to alum. Monkeys receiving placebo served as controls. The monkeys received a total of 4 doses of vaccine with the first 3 doses given every 3 weeks and the last booster dose administered 14 weeks later. The monkeys were challenged orally with H. pylori one week after the last vaccine dose and euthanized 10 weeks after challenge, at which time, their stomachs were collected for determination of bacterial colonization and histopathology. Monkeys primed with the oral vaccine and boosted with the parenteral vaccine showed a statistically significant reduction in bacterial colonization when compared to sham-immunized control animals (P = 0.05; Wilcoxon rank sums test). Monkeys receiving parenteral only regimes of urease plus Bay or alum showed no difference in bacterial colonization compared with sham-immunized controls (P = 1.00 and P = 0.33, respectively). The mucosal prime parenteral boost regime did not cause gastropathy. There was no difference in any of the 3 treatment groups with respect to gastric epithelial changes compared to control animals. There was also no difference in the type and extent of gastric inflammatory cell infiltrates between animals vaccinated by the mucosal prime parenteral boost strategy and sham immunized controls. However, monkeys receiving the two parenteral-only regimens had slightly elevated gastritis scores. PMID- 10462243 TI - Response to HBV vaccine in relation to anti-HCV and anti-HBc positivity: a study in intravenous drug addicts. AB - Drug addicts represent the group of young adults with the lowest response to hepatitis B virus (HBV) vaccine. A study was carried out on 110 current intravenous heroin users attending the service providing assistance to intravenous drug users (IVDUs) (SERT) in Padua: 66.4% of them were found anti hepatitis C virus (HCV)-positive and 33.6% were anti-HBc positive; 29.9% were positive for both. The subjects were vaccinated with 10 microg of yeast-derived vaccine at months 0, 1 and 2 (fast schedule). The overall response rate was 66.4%. Response seems to be affected by positivity to anti-HBc, but not to HCV infection. PMID- 10462244 TI - Immune response to revaccination with meningococcal A and C polysaccharides in Gambian children following repeated immunisation during early childhood. AB - Forty-two Gambian children randomised to receive two doses of meningococcal A/C polysaccharide vaccine (MPS) in infancy and either MPS (n = 15), meningococcal A/C conjugate (n = 13) or inactivated polio vaccine (IPV n = 14) at 2 years, were revaccinated with MPS at 5 years of age along with 39 matched control children. Meningococcal A and C polysaccharide antibodies were analysed by ELISA and bactericidal assay (SBA) in sera taken before and 10 days after revaccination. The geometric mean group SBA titre in the MPS group following revaccination was about half that of the unvaccinated controls (0.51 95%CI: 0.28, 0.90) for group A and less than half that of the controls for group C (0.41, 95%CI: 0.16, 1.03 P = 0.06). The group C SBA response in the conjugate group was 14-fold higher than in the MPS group (P < 0.001). Multiple doses of meningococcal polysaccharide in childhood may therefore attenuate the SBA response to both group A and group C polysaccharides. In contrast, vaccination with meningococcal A/C conjugate after MPS in infancy gives immunological memory to N. meningitidis group C. PMID- 10462245 TI - Safety and immunogenicity of MF59-adjuvanted influenza vaccine in the elderly. AB - Safety and immunogenicity of the influenza vaccine adjuvanted with MF59 (FLUAD) were compared to those of a non adjuvanted subunit vaccine in elderly subjects during three consecutive influenza seasons. Geometric mean titres and proportions of subjects with either a > or = four-fold increase in antibody titres or with an HI titre > or = 128 after immunisation were higher in FLUAD recipients. The adjuvant effect on the magnitude of the responses was most pronounced in subjects with pre-vaccination titres < or = 40. Although associated with more frequent mild local reactions, the adjuvanted vaccine was well tolerated. Thus, the addition of MF59 increased the immunogenicity of the subunit influenza vaccine in elderly persons with low pre-vaccination titres, who are at greatest risk of developing severe influenza disease and vaccine failure, without a clinically important increase in reactogenicity. PMID- 10462246 TI - Oral immunisation of mice with live Japanese encephalitis virus induces a protective immune response. AB - This study was undertaken to evaluate the efficacy of oral immunisation of mice with live Japanese encephalitis virus (JEV). Swiss albino mice were immunised with JEV by the peroral (p.o.), intraperitoneal (i.p.) and the subcutaneous (s.c.) routes on days 0, 7 and 14 using either mouse brain derived immunogen (MBDI) or cell culture derived immunogen (CCDI). Oral immunisation of mice evoked high anti-JEV antibody titres by ELISA (Geometric mean titres of 5065 with CCDI and 8854 with MBDI). Moreover, the orally immunised mice showed 76.7% protection with MBDI and 70% with CCDI against intracerebral challenge with a lethal dose of JEV. This study demonstrates for the first time that oral immunisation of mice with live JEV generates a brisk, protective immune response. The results of this study suggest that oral immunisation with JEV holds promise for the future. PMID- 10462247 TI - Quantitative analysis of IgG class and subclass and IgA serum response to Shigella sonnei and Shigella flexneri 2a polysaccharides following vaccination with Shigella conjugate vaccines. AB - It has been recently reported that a conjugate vaccine composed of the O-specific polysaccharide of S. sonnei bound to Pseudomonas aeruginosa recombinant exoprotein A (rEPA) conferred 74% protection against S. sonnei shigellosis. In the present study affinity purified Shigella antibodies were used as standards to quantify and characterize the serum antibody response to vaccination with Shigella sonnei or Shigella flexneri 2a polysaccharide conjugated to rEPA. The geometric mean concentrations of antibodies at the pre-vaccination stage were 3.8 microg/ml for IgG anti-S. sonnei LPS and 11.26 microg/ml for IgG anti-S. flexneri 2a LPS. Vaccination with S. sonnei-rEPA and S. flexneri 2a-rEPA induced the production of specific IgG antibodies to levels of 115.8 microg/ml and 126.5 microg/ml, respectively. The levels of specific antibodies above the pre vaccination values persisted for at least 2 years. The IgG response to S. flexneri 2a-rEPA conjugate was almost entirely represented by the IgG2 subclass. The concentration of IgG1 anti-S. sonnei LPS was significantly higher than that of IgG2 14 days after vaccination with the homologous conjugate, but decreased to similar levels to those of IgG2 6, 12 and 24 months after immunization. Since the only difference between the S. sonnei and S. flexneri 2a conjugates lies in the different polysaccharides of the two Shigella serogroups (the protein rEPA, is identical in both cases), it follows that the different pattern of IgG subclass response is a result of the different structures of the two O-polysaccharides of S. sonnei and S. flexneri 2a. PMID- 10462248 TI - Prior immunity to a carrier enhances antibody responses to hCG in recipients of an hCG-carrier conjugate vaccine. AB - Pre-sensitization with carrier often leads to epitopic suppression of subsequent anti-hapten antibody responses. To ascertain whether epitopic suppression occurs in humans, we examined the effect of pre-existing anti-carrier immunity on antibody responses to hCG in volunteers of a clinical trial of an hCG-based conjugate birth-control vaccine. When we studied the correlation between pre vaccination anti-carrier immunity and post-vaccination anti-hCG responses, we found that prior immunity to the carriers did not lead to epitopic suppression of anti-hCG responses. On the contrary, it was found that prior immunity to TT, one of the two carriers used in this vaccine, led to enhancement of anti-hCG responses. Our data indicates that prior immunity to the carriers may not be detrimental to the performance of conjugate vaccines, and may actually be beneficial in some cases. PMID- 10462249 TI - Venezuelan equine encephalitis virus vectors expressing HIV-1 proteins: vector design strategies for improved vaccine efficacy. AB - A live virus vaccine vector has been constructed from a molecularly cloned attenuated strain of Venezuelan equine encephalitis virus (VEE). High levels of foreign protein expression are regulated by an additional copy of the 26 S viral subgenomic RNA promoter. The position of this additional promoter and foreign gene in the VEE genome was predicted to have a major influence on expression level of the heterologous protein. Two sites in the genome were tested to determine the optimal site for expression of the matrix/capsid (MA/CA) coding region of human immunodeficiency virus (HIV-1). One vector contained the additional promoter and the MA/CA genes immediately downstream of the VEE E1 gene at the 3' end of the genome. In the second vector, the additional promoter was introduced immediately upstream from the authentic 26 S subgenomic promoter. Significantly higher levels of MA/CA were expressed from the downstream vector compared to the upstream vector. However, the stability of expression for both vectors was similar following passage in baby hamster kidney cells (BHK) cells. In BALB/c mice, the two vectors elicited similar levels of cellular immune responses to MA/CA as determined by bulk cytotoxic T-lymphocyte assays and precursor frequency analysis, but the humoral response induced by the downstream vector was significantly stronger. At 11 months post boosting with the downstream vector, serum antibody levels against HIV MA/CA were undiminished, and MA/CA specific CTLp were detectable in all mice tested. These findings suggest that VEE vectors can be optimized to elicit strong, balanced and long-lived immune responses to foreign viral proteins. PMID- 10462250 TI - Construction and immunogenicity of DNA vaccine plasmids encoding four Plasmodium vivax candidate vaccine antigens. AB - Plasmodium vivax is the second most common agent of human malaria. Although infection is rarely fatal, it nonetheless imposes a significant burden of illness in endemic areas. A successful vaccine against P. vivax will likely need to induce immune responses against both pre-erythrocytic and erythrocytic stage forms of the parasite. Accordingly, we constructed eight nucleic acid vaccines based on four antigens, the circumsporozoite protein (PvCSP) and sporozoite surface protein 2 (PvSSP2) from the pre-erythrocytic stage, and apical membrane antigen 1 (PvAMA1) and merozoite surface protein 1 (PvMSP1) from the erythrocytic stage. The constructs induced high levels of specific antibody in mice regardless of whether the antigen was expressed in native form or fused to a human tissue plasminogen activator leader peptide. High titer antibodies induced against PvCSP did not react with the protective AGDR epitope within the sequence of this antigen. These results support the immunogenicity of these four vaccine candidate antigens when delivered as nucleic acid vaccines. PMID- 10462251 TI - Human phase I vaccine trials of 3 recombinant asexual stage malaria antigens with Montanide ISA720 adjuvant. AB - Two phase I vaccine trials were conducted to test the immunogenicity and safety of a vaccine containing three recombinant malaria antigens from the asexual stage of Plasmodium falciparum. The three antigens are a fragment of MSP1 (190LCS.T3); MSP2 and a portion of RESA and were formulated in Montanide ISA720 adjuvant. These trials investigated the dose response of each antigen for eliciting both antibody and T-cell responses and the immunogenicity of a mixture of the antigens compared with the antigens injected separately. All three antigens elicited both antibody and T-cell responses. Strong T-cell responses were observed with 190LCS.T3 and RESA with stimulation indices exceeding 100 for peripheral blood leucocytes in some individuals. The antibody responses were generally weak. The human antibody responses observed with MSP2 in Montanide ISA720 were not significantly different from those obtained in an earlier trial which used MSP2 with alum as the adjuvant. No antigenic competition was observed: volunteers receiving a mixture of antigens had similar responses to those receiving the three antigens at separate sites. Tenderness and pain at the injection site were common over the first few days following immunization. In some volunteers, especially those receiving the highest doses tested, there was a delayed reaction at the injection site with pain and swelling occurring approximately 10 days after injection. PMID- 10462252 TI - Characterization of the humoral and cellular immune responses against hepatitis C virus core induced by DNA-based immunization. AB - Hepatitis C Virus (HCV) causes most cases of posttransfusion hepatitis. Chronic HCV infection is highly related to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Current therapies are only minimally effective and no vaccine has been developed. DNA-based immunization could be of prophylactic and therapeutic value for HCV infection. By intramuscular inoculation in BALB/c mice with an HCV recombinant plasmid pCI-HCV-C, we found significant levels of IgM antibody, but no significant IgG rise. After boost the immunized mice with recombinant HCV-core protein (cp1-10; 1-164aa), the anticore IgG, verified by Western-blotting, rose rapidly, which was two weeks earlier than that with control plasmid. Spleen cells from pCI-HCV-C immunized mice gave higher proliferation index (PI) than control (P < 0.05). The PI of cp1-10 boosted mice was even higher. Proliferation blocking assay with mAb proved the responding cell to be of CD4+ CD8- phenotype, supporting specific priming of T helper cells. A 51Cr-releasing CTL assay specific for HCV-core was developed, and a specific CTL response against HCV-core was demonstrated in both pCI-HCV-C immunized mice and mice boosted with cp1-10. Strong cytotoxic activity against peptide-pulsed p815 cells (H-2d), but not EL-4 cells (H-2b), suggested MHC class I restriction of the CTL activity. Blocking of CTL with mAb proved the effector cells to be of CD4- CD8+. Three CTL epitopes in HCV-core protein were demonstrated. We failed to detect CTL when immunized only with core protein. The results suggested that vaccination with HCV-core derived DNA sequences could be an effective method to induce humoral and cellular immune responses to HCV. PMID- 10462253 TI - Immunity obtained by gene-gun inoculation of a rotavirus DNA vaccine to the abdominal epidermis or anorectal epithelium. AB - We have previously shown that gene-gun delivery of murine rotavirus DNA vaccines to the epidermis induced protection against rotavirus challenge in mice. In this study, we used a rotavirus group antigen (VP6)-specific DNA vaccine to compare epidermal immunization with immunization to the anorectal epithelium for efficacy in inducing protective immunity. The vaccine was administered into cells of the abdominal epidermis or anorectal epithelium of adult BALB/c mice with an Accell gene-gun (PowderJect, Inc). Vaccines administered by either route elicited rotavirus-specific ELISA antibodies and analysis of the IgG subtypes indicated Th2-type responses were generated by both routes of administration, in contrast to Th1-type responses generated by live rotavirus. Protection against virus challenge was obtained in mice inoculated by either route, as shown by significant reduction of virus excreted in stools. The protection obtained by immunization of the anorectal epithelium was greater than that for epidermal immunization at the same vaccine dose. These results suggest that mucosal immunization of DNA vaccines may be an effective means to generate protective immunity against mucosal pathogens. PMID- 10462254 TI - The Fas ligand/Fas system in renal injury. AB - The FasL-Fas system regulates renal cell apoptosis, as well as the immune and inflammatory responses. Evidence that FasL and Fas participate in renal injury may be summarized along modified Koch's postulates (Table 1): (i) FasL is expressed by renal cells and during renal injury, (ii) activation of the Fas receptor promotes apoptosis of cultured renal cells, (iii) Fas agonists induce glomerular injury but they may also decrease renal injury by limiting injurious immunological responses, (iv) mice with disrupted FasL/Fas systems are protected from tubular cell injury during ischaemia reperfusion, although they develop autoimmune glomerulonephritis if other genetic predisposing factors are present. FasL/Fas must be considered a new target for therapeutic intervention in renal injury. Therapeutic modulation of Fas should aim not only at protecting intrinsic glomerular or tubular epithelial cells from death, but also at modulating the immune, inflammatory, and fibrogenic responses. Possible therapeutic interventions include Fas agonists, soluble Fas receptors, or other antagonists, and targetting of Fas to undesired cells, such as fibroblasts, in order to decrease their numbers in a physiological manner through apoptosis. Any therapeutic attempt should carefully take into account the possible effects of interference with Fas in other 1833 cell systems. Given the complexities of the FasL/Fas system, further studies are warranted. PMID- 10462255 TI - Antithrombotic measures for indwelling intravenous haemodialysis catheters- Columbus' egg yet to be found. PMID- 10462256 TI - The role of ACE inhibitors and angiotensin II receptor blockers in the response to epoetin. PMID- 10462257 TI - Prevention of renal function loss after non-renal solid organ transplantation- how can nephrologists help to keep the kidneys out of the line of fire? PMID- 10462258 TI - Hantavirus infection--haemorrhagic fever in the Balkans--potential nephrological hazards in the Kosovo war. PMID- 10462259 TI - International nephrological assistance to Kosovo refugees. European Branch of the Task Force. PMID- 10462260 TI - Identification of non-diabetic glomerular disease in renal biopsies from diabetics--a dilemma. PMID- 10462261 TI - Lethal air embolism following removal of a double lumen jugular vein catheter. PMID- 10462262 TI - The growth hormone-insulin-like growth factor axis in kidney re-revisited. AB - The use of renal allotransplantation to treat ESRD in the US is limited by lack of organ availability. A possible solution is the transplantation of developing kidneys (metanephric allograft or xenografts). We have conducted studies that demonstrate the feasibility of such a strategy and have shown that IGF I may be useful to accelerate the growth and development of these transplanted organs. The rationale for the use of IGF I in this setting grew from a basic understanding of the role that the growth factor plays in kidney development. ARF in humans is the most costly kidney-related disease requiring hospitalization. Its incidence is increasing. Despite many advances in dialytic therapy, the mortality rate for patients with ARF has not changed in the last several decades. Strategies for treatment of ARF are directed toward supportive care to permit renal regeneration to occur. There exists a need for new therapeutic approaches that can speed recovery and reduce mortality. Although IGF I may not prove to be the 'magic bullet' for ARF, its proposal and testing as a potential therapeutic agent has provided a paradigm for the development of treatment modalities to accelerate renal regeneration based upon a basic understanding of the injury/repair process. The basis for development of a 'growth factor' therapy for ARF will probably evolve, at least in part, out of the testing and use of IGF I in rat models and in humans. The use of GH to treat ESRD was proposed shortly after its isolation and the demonstration of its action in increasing the rate of glomerular filtration. Later, it was discovered that the actions of GH on kidney are mediated by IGF I, and the means by which IGF I enhances glomerular filtration was elucidated. We have shown that humans with ESRD are not resistant to the actions of IGF I in enhancing the GFR, establishing the potential for use of IGF I as a pharmacological agent for ESRD. There is no effective drug therapy to enhance renal function in ESRD. Although much work remains to be done, and clearly caution is advised, our observations establish the potential for the use of IGF I as a therapeutic agent in this setting and justify continued study of IGF I as a medical therapy to delay the need for dialysis. PMID- 10462263 TI - Alphav integrin regulates TNF-alpha-induced nitric oxide synthesis in rat mesangial cells--possible role of osteopontin. AB - BACKGROUND: Tumour necrosis factor-alpha (TNF-alpha) induces nitric oxide (NO) synthesis in rat mesangial cells (MCs). We previously demonstrated that osteopontin (OP), a matrix protein that mainly interacts with the alphav integrin family, increased time-dependently by TNF-alpha stimulation at gene and protein levels. The regulation of NO synthesis by integrins or matrix proteins is unclear. METHODS: We examined whether integrin, especially alphav integrin, regulates NO synthesis in rat MCs and whether OP, an alphav integrin ligand, has an effect on TNF-alpha-induced NO synthesis. Furthermore, OP and inducible NO synthase (iNOS) gene expression was examined by Northern blotting. RESULTS: TNF alpha increased NO synthesis in MCs in a time-dependent manner. Synthetic GRGDSP peptide, which is known to inhibit various integrins that interact with RGD containing extracellular matrices, increased TNF-alpha-induced NO levels in a dose-dependent manner. Cyclical RGD peptide, the specific inhibitor of alphav integrin, also exhibited a dose-dependent effect of increasing NO levels, while GRGESP peptide, which has very low affinity to integrins, had no effect. In addition, NO synthesis was found to be significantly reduced when MCs were plated on OP-coated dishes compared to type I or IV collagen-coated dishes. Furthermore, anti-OP antibody increased NO synthesis in MCs. iNOS mRNA levels were increased by TNF-alpha, and were abruptly diminished after OP mRNA was significantly induced. CONCLUSIONS: The present study demonstrated the involvement of alphav integrin in TNF-alpha-induced NO synthesis in rat MCs, and the possible role of OP was suggested in the mechanism. TNF-alpha and extracellular matrices can co operate to regulate the behaviour of MCs at least partly through NO synthesis, which may participate in the course of glomerular diseases. PMID- 10462264 TI - Erythrogenic toxin type B and its precursor isolated from nephritogenic streptococci induce leukocyte infiltration in normal rat kidneys. AB - BACKGROUND: Leukocyte infiltration is a common feature in renal biopsies from patients with acute poststreptococcal glomerulonephritis (APSGN). Cationic streptococcal erythrogenic toxin type B (ETB) and its precursor (ETBP) have been implicated in the pathogenesis of the disease, and the presence of ETB has been evidenced in renal biopsies from patients with APSGN. The present studies were performed to determine the effect of the ETBP and ETB on renal leukocyte infiltration and the mechanism(s) implicated in the phenomenon. METHODS: Male Sprague-Dawley rats were injected intrarenally with 100 microg of ETB or ETBP. Animals were sacrificed at 1, 6 and 24 h after injection and renal samples were studied by indirect immunofluorescence for the presence of leukocyte common antigen (LCA+) cells, C3, monocyte chemotactic protein-1 (MCP-1) and intercellular adhesion molecule-(ICAM-1), and by direct immunofluorescence for the presence of immunoglobulins. ETB and ETBP were tested for chemotactic effect and migration inhibition factor (MIF) activity by chemotaxis under agarose and agarose microdroplet methods, respectively. Streptococcal proteins were also tested for the capacity to induce MIF activity in rat glomerular cultures. To test for the influence of cationic charge on renal LCA+ cell infiltration, rats were injected with cationized ferritin or polyethyleneimine (PEI) and sacrificed 1 h later. RESULTS: An increased number of LCA+ cells was found in glomeruli and interstitial areas in ETB- or ETBP-injected animals. ETB and ETBP showed chemotactic and MIF activity on neutrophils and macrophages, and ETBP induced MIF activity in supernatants of glomerular cultures. Data obtained from C3, MCP-1, ICAM-1 or immunoglobulin renal staining in experimental animals were not significantly different when compared to control values. Cationized compounds failed to induce LCA+ cell infiltration; however, an increased number of glomerular LCA+ cells was observed after PEI perfusion. CONCLUSIONS: ETB and ETBP induce renal LCA+ cell infiltration during a short period after intrarenal injection, and this finding could be mediated by chemotactic and MIF activities. These observations could be relevant in the early events of pathogenesis of APSGN. PMID- 10462265 TI - IgA-gliadin antibodies, IgA-containing circulating immune complexes, and IgA glomerular deposits in wasting marmoset syndrome. AB - BACKGROUND: Marmosets in captivity are highly susceptible to wasting marmoset syndrome (WMS), the aetiology of which is still not fully determined. METHODS: The level of IgA-gliadin antibodies (IgA-AGA), of IgA-containing circulating immune complexes (IgA-CIC), and the degree of glomerular IgA deposits were compared between marmosets suffering from WMS and animals not affected by the disorder. RESULTS: Both IgA-AGA and IgA-CIC were demonstrable in all groups of monkeys investigated. IgA-AGA and IgA-CIC were significantly higher in monkeys with WMS than in non-affected animals. There was a significant correlation between the glomerular IgA-deposition and titre of IgA-AGA. The group of marmosets strongly positive for glomerular IgA deposits comprised significantly more animals suffering from WMS than the group without deposits. In the diet of the animals a considerable amount of gliadin-like cereal proteins was assayed. CONCLUSIONS: There are several parallels between the human disorders (coeliac disease and IgA-nephropathy/Berger's disease) and the changes observed in WMS. It should be further investigated if WMS in marmosets is a suitable animal model for both human diseases. PMID- 10462266 TI - Endothelin ET(A) receptor blockade prevents the progression of renal failure and hypertension in uraemic rats. AB - BACKGROUND: Elevated plasma and urine endothelin-1 (ET-1) levels have been reported in renal failure and may be involved in renal disease progression. We investigated whether these changes are related to increased vascular and renal ET 1 production in the pole resection remnant kidney model of chronic renal failure in the rat. METHODS: Uraemic Wistar rats were prepared by surgical renal mass 5/6 ablation and compared with sham-operated controls (protocol 1). Immunoreactive-ET 1 (ir-ET-1) concentration was measured by radioimmunoassay after sample extraction and purification. To investigate the functional role of ET-1 during the progression of chronic renal failure, uraemic rats (protocol 2) were treated with either the vehicle or the ET-1 type A (ET(A)) receptor antagonist LU135252 (LU). RESULTS: Systolic blood pressure and serum creatinine, as well as urinary volume and proteinuria, were significantly higher, whereas creatinine clearance was reduced in uraemic rats compared with sham-operated controls. As expected, plasma and urine ir-ET-1 concentrations were increased in uraemic rats (P<0.01) and were related to the increased ir-ET-1 levels in blood vessels and glomeruli (P<0.001). Positive correlation was found between plasma, thoracic aorta and mesenteric arterial bed ir-ET-1 levels and systolic blood pressure, as well as blood vessel hypertrophy. In addition, increased urinary ir-ET-1 excretion correlated with the rise in serum creatinine and proteinuria. In protocol 2, a 3 week treatment period with LU was initiated once uraemia and hypertension were established. In untreated uraemic rats, systolic blood pressure increased further (P<0.05), but this was not the case in LU-treated uraemic rats. At the end of treatment, serum creatinine and proteinuria were significantly lower (P<0.05) and creatinine clearance was higher (P<0.01) in LU-treated rats compared with uraemic untreated animals. While plasma ir-ET-1 concentration was similar in the two groups, ir-ET-1 concentration in thoracic aorta, mesenteric arterial bed, renal cortex and urine was significantly lower in LU-treated animals (P<0.01). In addition, heart, thoracic aorta and mesenteric arterial wet weight to body weight ratios were also significantly reduced in LU-treated uraemic rats (P<0.05). CONCLUSIONS: Elevated plasma ET-1 concentration and urinary ET-1 excretion in rats with renal mass ablation are related to enhanced ET-1 production in vascular and renal tissues, thus suggesting an important role for ET-1 in the aggravation of hypertension and vascular hypertrophy as well as in the progression of renal insufficiency. These pathophysiological effects are prevented by treatment with selective ET(A) receptor blockade. PMID- 10462267 TI - The epidemiology and prognosis of glomerulonephritis in Denmark 1985-1997. AB - BACKGROUND: The existence of a national renal biopsy register and a national terminal uraemia status register in Denmark provides an opportunity to study the prognosis of glomerulonephritis (GN), and factors influencing prognosis. METHODS: Multivariate analysis of 2380 renal biopsies with GN performed between 1985 and 1997 was done to determine the influence of clinical and histological factors on prognosis. RESULTS: The incidence of GN (39/mo/year) and individual diagnoses did not change during the period. After 10 years, 32%, were dead, 13% terminally uraemic, 5%, uraemic and 50% well. Older age increased mortality, but not the incidence of renal failure after the first year. Male sex increased both mortality and incidence of renal failure (34 vs 24% at 10 years, P < 0.001). The diagnoses could be divided into three prognostic groups compared with the general population: a good prognostic group (minimal change GN and membranous GN ), with a relative mortality of three and a combined renal and patient mortality of four; a poor prognostic group [crescentic GN, HUS/TTP, chronic GN] with relative mortalities of 8-19 and 13-33, respectively; and the remainder with mortalities of 4-7 and 6-12. The presence of multiple glomerular pathology, chronic GN, nephrosclerosis and chronic interstitial nephropathy worsened the prognosis, while the presence of immune deposits only worsened the prognosis of focal segmental glomerulopathy. Mortality was related to uraemia and co-morbidity at biopsy, and to the incidence of renal failure. Renal failure was correlated to uraemia and hypertension at biopsy but not to nephrotic syndrome or atherosclerosis. All vascular complications were increased and were positively related to hypertension and negatively correlated to the incidence of uraemia. Crescentric glomerulonephritis combined with anti-GBM disease had a worse prognosis than Wegener's granulomatosis, with microscopic polyangiitis and pauci immune disease occupying an intermediate position. The prognosis of mesangioproliferative GN was unaffected by the presence of IgA nephropathy and systemic lupus erythematosus. PMID- 10462268 TI - In vivo microvascular clearance of albumin in renal and extrarenal tissues in puromycin aminonucleoside (PAN) induced nephrotic syndrome. AB - BACKGROUND: The nephrotic syndrome is characterized by generalized oedema considered to be due to the fall in serum albumin and to sodium retention. The aim of the present study was to investigate whether a generalized disturbance in vascular integrity contributes to the oedema formation. METHODS: We used the PAN (puromycin aminonucleoside) nephritis model in order to induce the nephrotic syndrome in female Wistar rats. Eight rats were given PAN, 15 mg/100 g body weight, intraperitoneally 10 days prior to the study, whereas 21 rats served as controls. Albumin clearance to tissues was measured using a dual isotope technique. Repeated blood samples as well as samples from various muscles, kidney, liver, lung, heart, abdominal wall and from ascites fluid were taken to determine radioactivity and tissue dry-to-wet weights. Clearance of albumin (Clalb) from plasma to interstitium was calculated from the (linear) increment in 'plasma equivalent tissue albumin space' as a function of time, corrected for intravascular volume and oedema. The plasma and urine concentrations of albumin were determined in a parallel study by single radial diffusion using monospecific rabbit anti-rat antiserum in seven PAN animals and 13 controls. RESULTS: A marked fall in dry-to-wet weight ratios together with pronounced proteinuria, oedema and ascites were found in the PAN animals. Haematocrit decreased from 45% (32-51) to 30% (28-38) and serum albumin from 22.0 g/l (16.3-25.2) to 4.94 g/l (3.20-6.72) in control and PAN animals, respectively. However, Clalb apparently remained unchanged in the PAN animals in comparison to controls in most tissues examined. Thus, in these in vivo experiments there was no direct evidence of an increased extravasation of albumin in extrarenal tissues. CONCLUSIONS: There was no strong support for the contention that a generalized disturbance of capillary integrity outside the renal vasculature would contribute to the oedema formation in the PAN nephrotic syndrome. PMID- 10462269 TI - Plasma renin and prorenin and renin gene variation in patients with insulin dependent diabetes mellitus and nephropathy. AB - BACKGROUND: The most striking abnormality in the renin angiotensin system in diabetic nephropathy (DN) is increased plasma prorenin. Renin is thought to be low or normal in DN. In spite of altered (pro)renin regulation the renin gene has not been studied for contribution to the development of DN. METHODS: We studied plasma renin, prorenin, and four polymorphic markers of the renin gene in 199 patients with IDDM and DN, and in 192 normoalbuminuric IDDM controls matched for age, sex, and duration of diabetes. Plasma renin and total renin were measured by immunoradiometric assays. Genotyping was PCR-based. RESULTS: Plasma renin was increased in patients with nephropathy (median (range), 26.3 (5.2-243.3) vs 18.3 (4.2-373.5) microU/ml in the normoalbuminuric group, P<0.0001). Prorenin levels were elevated out of proportion to renin levels in nephropathic patients (789 (88 5481) vs 302 (36-2226) microU/ml, P<0.0001). Proliferative retinopathy had an additive effect on plasma prorenin, but not on renin. DN was associated with a BglI RFLP in the first intron of the renin gene (bb-genotype: n=106 vs 82 in DN and normoalbuminuric patients respectively, P=0.037), but not with three other polymorphisms in the renin gene. A trend for association of higher prorenin levels with the DN-associated allele of this renin polymorphism was observed in a subgroup of patients with DN (bb vs Bb+BB, P=0.07). CONCLUSIONS: The results indicate that in DN there is an increase in both renin and prorenin levels. A renin gene polymorphism may contribute weakly to DN. Although speculative, one of the renin gene alleles could lead to increased renin gene expression, leading to higher renin and prorenin levels. These may play a role in the pathogenesis of DN. PMID- 10462270 TI - The impact of withdrawing ACE inhibitors on erythropoietin responsiveness and left ventricular hypertrophy in haemodialysis patients. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inhibitors have the capability of decreasing left ventricular mass index (LVMI) in chronic haemodialysis (HD) patients. On the other hand, recent reports provide conflicting information regarding the impact of ACE inhibitors on responsiveness to recombinant human erythropoietin (rHuEpo), and there are no data about the effect of withdrawing ACE inhibitors both on rHuEpo response and LVMI in HD patients. METHODS: ACE inhibitors were switched to another antihypertensive medication in 23 out of 68 patients in our HD unit who were receiving both rHuEpo and an ACE inhibitor for more than 1 year. Blood pressure at the pre- and post-dialysis phases, haematocrit levels and rHuEpo doses were determined at the end of the first and of the third years, and the LVMI was determined at the end of the third year. Statistical analyses were done in 15 patients in whom the study could be completed. RESULTS: The mean (+/-SD) haematocrit level was increased from 26.3+6.4% to 29.8+/-6.3% at the first year (P<0.05), and to 29.4+/-6.5% at the third year (P<0.05 vs before), while the mean dose of rHuEpo was decreased from 208.3+/-99.0 UI/kg/week to 141.0+/-91.8 at the first year (P=0.01), and to 141.4+/-81.0 at the third year (P=0.01 vs before). Administration of rHuEpo had been stopped in two patients at the end of the first year. The mean blood pressure level and the mean LVMI were not changed (P>0.05 vs before). There were no significant changes in dialysis parameters, iron status, plasma renin activities, and levels of aldosterone, intact parathyroid hormone, aluminum and erythropoietin. CONCLUSION: The findings of this small uncontrolled study indicate that withdrawal of ACE inhibitors in hypertensive chronic HD patients receiving rHuEpo may result in an increase in haematocrit level, and a decrease in dose of rHuEpo without any significant changes in the blood pressure level and LVMI. Controlled prospective studies are needed to clarify this issue. PMID- 10462271 TI - Usefulness of quantitative heel ultrasound compared with dual-energy X-ray absorptiometry in determining bone mineral density in chronic haemodialysis patients. AB - BACKGROUND: Reduced bone mineral density (BMD) is associated with renal osteodystrophy and osteoporosis in end-stage renal failure patients. Dual-energy X-ray absorptiometry (DXA) is the standard non-invasive method to assess BMD, but is not always widely available. Quantitative heel ultrasound (QUS) is a mobile, relatively inexpensive, easy to perform and radiation-free method which can predict fractures to the same extent as DXA. This study assessed the usefulness of QUS vs DXA in determining BMD in chronic haemodialysis patients. METHODS: Patients had their BMD at the hip and spine measured by DXA (Lunar Expert). QUS of the left heel (McCue CubaClinical II machine) measured broadband ultrasound attenuation (BUA) and velocity of sound (VOS). Correlations between DXA and QUS parameters were calculated. Receiver operator characteristic (ROC) curves were plotted for BUA and VOS and used to define cut-off points for calculating sensitivities and specificities for BUA and VOS. Femoral neck BMD was applied as the standard for diagnosing osteoporosis (T< or =-2.5) and osteopaenia (T>-2.5 and < or =-1) by WHO criteria. RESULTS: Eighty eight patients (45.5% women), mean age 58+/-17 years, were studied. A total of 19% and 49% had femoral neck BMDs in the 'osteoporosis' and 'osteopaenia' ranges, respectively. There were good correlations between hip BMD and QUS parameters (r=0.68-0.79, P<0.001). Areas under the ROC curves for BUA and VOS in diagnosing 'osteoporosis' were 0.86 and 0.80, respectively. BUA and VOS had sensitivities of 76 and 71% and specificities of 80 and 69%, respectively, for diagnosing 'osteoporosis'. The positive predictive values for BUA and VOS were 48 and 35%, respectively, and the negative predictive values were 93 and 91% respectively. CONCLUSIONS: DXA and QUS parameters were significantly correlated. However, sensitivities and specificities of QUS parameters were not sufficiently high for QUS to be used simply as an alternative to DXA. The relatively high negative predictive values suggest that QUS may reliably screen out patients unlikely to have a BMD in the osteoporotic range. The relatively low positive predictive values, however, mean that subjects classified as osteoporotic using QUS require further investigations such as DXA to confirm the diagnosis. PMID- 10462272 TI - Risk factors for reduced bone density in haemodialysis patients. AB - BACKGROUND: Renal osteodystrophy may result in considerable morbidity for patients with end-stage renal disease. Secondary hyperparathyroidism, adynamic bone disease and osteomalacia, the main bony problems in chronic renal failure, may all be responsible for a reduction in bone mineral density (BMD). This can result in an increased fracture risk. By virtue of their age, post-menopausal status (in women), sedentary life-style and treatment (including previous corticosteroids), haemodialysis patients may be expected also to be at risk for developing osteoporosis, but little is known about the relative importance of these factors. METHODS: We report a prospective study examining the prevalence of reduced bone mineral density (BMD) and its association with a wide range of factors, in a heterogenous group of 88 chronic haemodialysis patients. Femoral neck and lumbar BMD were measured by dual-energy X-ray absorptiometry (DXA). Stepwise multiple linear regression analysis was used to identify risk factors associated with low bone mass. RESULTS: Forty three patients (48.9%) had reduced BMD, and in 17 (19.3%) BMD was below the fracture threshold as defined on DXA measurements by the World Health Organization (WHO). The BMD had significant negative associations with age, serum parathyroid hormone (PTH) levels, current gastric acid suppression therapy, female gender, age at menarche and history of previous fracture. Positive associations were found with weight, haemoglobin concentration, average serum phosphate, weekly heparin dose, oral calcium supplementation and history of parathyroidectomy. CONCLUSIONS: We have confirmed the importance of PTH-related bone disease in affecting BMD in haemodialysis patients, but have found that some other factors, which are known to be risk factors for osteoporosis, are also important. PMID- 10462273 TI - Epidemiology of Helicobacter pylori in chronic haemodialysis patients using the new RIBA H. pylori SIA. AB - BACKGROUND: There are few data concerning the epidemiology of H. pylori in patients on chronic haemodialysis (HD) treatment. These surveys concerned small populations and were made with ELISA technique. However, ELISA-based assays do not differentiate between strains of H. pylori that are associated with ulcers. Recent literature reports that formation of ulcers correlates strongly with the expression of cytotoxin-associated protein (CagA) and vacuolating cytotoxin (VacA) of H. pylori. METHODS: A novel serological test (RIBA H. pylori strip immunoblot assay (SIA)) has been recently introduced, it uses the H. pylori lysate (Lys) along with two additional purified recombinant antigens derived from CagA and VacA of H. pylori. AIM: To study the epidemiology of H. pylori using RIBA H. pylori SIA among chronic HD patients and blood donors as a control group. In addition, the activity of H. pylori was analysed by immunoblot technique in a group of patients with documented ulcers and normal renal function. RESULTS: The prevalence of antibody towards H. pylori among HD patients, blood donors, and patients with documented ulcers was 56% (127/228), 53% (84/158), and 100%, (21/21) respectively; the difference was significant (P=0.0001). The frequency of anti-H. pylori-positive individuals was significantly higher in patients with documented ulcers than HD patients and blood donors, 21/21 (100%) vs 211/386 (55%), P=0.0001. The frequency of antibody to H. pylori in the HD population was significantly associated with race (P= 0.005); no relationship between anti-H. pylori antibody and numerous demographic, biochemical, and clinical features of patients was seen. The frequency of antibodies against virulent strains of H. pylori in HD patients and blood donors with H. pylori was 60% (76/127) and 61% (51/84) respectively; it was 86% (18/21) among individuals with documented ulcers. No significant difference among these three groups occurred. CONCLUSIONS: The frequency of antibody towards H. pylori by RIBA H. pylori SIA was high both in HD patients and blood donors; patients with documented ulcers and normal renal function had significantly higher frequency of anti-H. pylori antibody. The anti H. pylori antibody rate among HD patients was strongly associated with race. The prevalence of antibody against virulent strains of H. pylori did not change among HD patients and control groups. Studies in large cohorts of HD patients with documented peptic ulcer disease are in progress. PMID- 10462274 TI - Cardiovascular morbidity and endothelial dysfunction in chronic haemodialysis patients: is homocyst(e)ine the missing link? AB - BACKGROUND: Haemodialysis patients exhibit an excessive burden of atherothrombotic disease, which is not explained adequately by traditional risk factors. Hyperhomocyst(e)inaemia, a consistent finding in uraemic patients, is now widely recognized as an independent risk factor for vascular disease. The aim of this study was to examine the hypothesis that hyperhomocyst(e)inaemia is associated with cardiovascular complications in dialysed patients. METHODS: In a cohort of 63 stable chronic haemodialysis patients, we examined the causal relationship between hyperhomocyst(e)inaemia and vascular endothelial and haemostatic function. All their markers were determined before and after an 8 week course of a 10 mg per day oral folate supplementation, a manoeuvre known to decrease hyperhomocyst(e)inaemia in uraemic patients. RESULTS: History of at least one cardiovascular atherothrombotic event was present in 47.6% of the haemodialysed patients, and radiographic evidence of vascular calcifications in 70%. Hyperhomocyst(e)inaemia was found in all patients, averaging 3.5-fold the upper limit of normal values (P<0.001), despite the lack of clinical and biological evidence of malnutrition. Fibrinogen, von Willebrand factor and plasminogen activator inhibitor type 1, but not endothelin 1, were significantly higher in haemodialysis patients than in controls. After adjustment for all variables, past history of cardiovascular events was independently associated with higher levels of homocyst(e)inaemia only (odds ratio (OR) 1.06; 95% confidence interval (CI) 1.01-1.12; P<0.026). The presence of aortic calcifications was independently and significantly associated with age (OR 1.37; 95% CI 1.07-1.75; P<0.025), homocyst(e)inaemia (OR 1.14; 95% CI 1.02-1.27; P<0.05) and fibrinogen concentration only (OR 9.74; 95% CI 1.25-75.2; P<0.05). None of the endothelial haemostatic factors was, however, related to homocyst(e)ine levels. Mid-term folate supplementation decreased plasma homocyst(e)ine levels significantly without achieving normal values. No significant change of endothelial-haemostatic markers was observed, however, despite the drop in plasma homocyst(e)ine. CONCLUSIONS: Hyperhomocyst(e)inaemia is associated with increased cardiovascular risk in haemodialysis patients. Folate supplementation was partially effective in lowering hyperhomocyst(e)inaemia, but its usefulness in terms of reduction in cardiovascular morbidity and mortality remains to be determined in prospective trials. PMID- 10462275 TI - A single dose of dalteparin effectively prevents clotting during haemodialysis. AB - BACKGROUND: A single bolus dose of LMW heparin at the start of haemodialysis effectively prevents clot formation in the dialyser and bubble trap. However, there are few studies on the appropriate dosage of LMW heparins in haemodialysis. Therefore we examined the relationship between the anticoagulant effect of dalteparin and clinical clotting during haemodialysis. METHODS: We performed an open, prospective study on the effect of decreasing doses of dalteparin in 12 haemodialysis patients during a total of 84 sessions (4-4.5 h). The normally applied dose of dalteparin in each patient was reduced by 25% for each session down to 50% of initial dose if no clotting was observed. Clinical clotting (grade 1-4) was evaluated by visual inspection after blood draining of the air trap every hour and by inspection of the dialyser after each session and compared to corresponding values for anti-FXa activity and dialysis time. Blood flow and ultrafiltration rate were kept within narrow limits throughout the study. RESULTS: No episodes of grade 4 clotting occurred, and no session was interrupted. Eighteen episodes of grade 3 clinical clotting (11%) were observed in patients without warfarin treatment, none with an anti-FXa activity >0.43 IU/ml. Oral warfarin treatment reduced the clinical clotting, and only one grade 3 episode was observed in patients on warfarin therapy. Anti-FXa activity and haemodialysis time were the only factors independently correlated to clotting in a logistic regression model. CONCLUSION: An anti-FXa activity above 0.4 IU/ml after 4 h of dialysis inhibits significant clotting during haemodialysis. A bolus dose of dalteparin of 70 IU/kg usually seems appropriate, but may be reduced in patients on warfarin treatment. Dialysis time is an independent risk factor for clinical clotting. PMID- 10462276 TI - Increased cardiac troponin T and endothelin-1 concentrations in dialysis patients may indicate heart disease. AB - BACKGROUND: Cardiac troponin T (cTnT) is a highly sensitive marker for the detection of myocardial damage. However, patients maintained on chronic dialysis often have increased serum cTnT concentrations without evidence of acute myocardial injury. The reason for this is unclear. In chronic haemodialysis patients, elevated plasma concentrations of big endothelin-1 (big ET-1) and endothelin-1 (ET-1) have been reported which may be associated with ischaemic heart disease. The aim of the present study was to investigate possible associations between cTnT, big ET-1, ET-1, other cardiac markers and cardiac disease in dialysis patients. METHODS: Thirty-six haemodialysis (HD) patients and 26 peritoneal dialysis (PD) patients without symptoms of acute myocardial ischaemia were investigated. In all patients, serum concentrations of cTnT (2nd generation ELISA), cardiac troponin I (TnI) (Opus, Behring), creatine kinase MB (CKMB) mass and creatine kinase (CK) were determined, in HD patients before and after dialysis. Additionally, in HD patients, plasma ET-1 and big ET-1 were measured. In 27 HD patients, left ventricular mass index (LVMI) was determined. Patients with ischaemic heart disease (IHD) were compared with non-IHD patients. RESULTS: Serum cTnT was elevated (> or =0.10 microg/l) in 20 of 36 HD patients and in eight of 26 PD patients. cTnI was elevated (> or =0.5 microg/l) in four of 62 dialysis patients. HD+PD patients with IHD showed higher cTnT than HD+PD patients without IHD, and ET-1 concentrations were higher in HD patients with than without IHD. In HD patients, there was a positive correlation between cTnT and big ET-1. In HD patients with left ventricular hypertrophy (LVH), serum cTnT, CKMB mass and post-dialysis plasma big ET-1 were higher than in patients with normal LVMI. Furthermore there was a positive correlation between cTnT levels and LVMI. CONCLUSION: These findings suggest that circulating cTnT may reflect left ventricular hypertrophy and/or myocardial ischaemia in dialysis patients, and indicate that ET-1 and big ET-1 might be associated with these conditions. PMID- 10462277 TI - Serum C-reactive protein (CRP) and risk of death in chronic dialysis patients. AB - BACKGROUND: The prognosis of chronic dialysis patients is poor, in part due to the high incidence of cardiovascular disease. Malnutrition, such as hypoalbuminaemia, has been shown to be a predictor of death in this group of patients, while serum C-reactive protein (CRP) is a predictor of myocardial infarction and sudden death. Thus, the aim of the present study was to determine of the relationship between CRP and serum albumin concentration, and the value of baseline CRP data in the prediction of death. METHODS: In one of the dialysis units in Okinawa, Japan, baseline CRP data was available (n=163, 95 men and 68 women) in January 1991. These patients were divided into two groups according to their baseline CRP levels, with group 1 consisting of CRP<10 mg/l (n=128) and group 2 of CRP> or =10 mg/l (n=35), and then followed up until the end of 1997. Survival curves were calculated using the Kaplan-Meier method. The statistical significance of the relationship between CRP levels and the risk of death was evaluated by multiple logistic analysis with covariables such as age, sex, diabetes mellitus, serum albumin, and blood pressure. RESULTS: The mean (SD) level of serum albumin was 38 (3) g/l in group 1 and 36 (3) g/l in group 2 (P<0.00001). The 5-year survival rate was significantly poorer in group 2 (44.4%) than in group 1 (82.5%) (P<0.0001). Furthermore, the risk of death was significantly higher in group 2 (relative risk 3.48 (95% confidence interval 1.76 6.89), P<0.0003) by multivariate Cox proportional hazard analysis. CONCLUSIONS: CRP is a significant predictor of death in chronic dialysis patients, independent of serum albumin and other possible confounders. Dialysis patients with high CRP levels should be carefully evaluated and monitored regardless of serum albumin concentrations in the normal range. PMID- 10462278 TI - The predictive value of cardiac troponin T measurements in subjects on regular haemodialysis. AB - BACKGROUND: Cardiac troponin T (cTnT) is a subunit of the cardiac actin-myosin complex, which leaks into the circulation when myocardial necrosis is present. Detection of cTnT is associated with a poor outcome in patients with unstable angina, and is a useful tool for risk stratification. The value of cTnT determination in patients with renal failure has been questioned, and the specificity of cTnT in this particular group has not been established. METHODS: In the present study, 94 patients at a single centre were followed prospectively after three determinations of cTNT, at 1-month intervals. The outcome after 12 months was chosen as the end-point. cTnT was measured using both a quantitative chemiluminiscence immunoassay and a qualitative rapid bedside immunoassay on a test strip. The maximum of three measurements was used and was correlated with different parameters and outcome. The following statistical tests were performed: Kaplan-Meier analysis, Cox's proportional regression analysis for measuring survival and logistic regression for analysing factors influencing cTnT. RESULTS: Forty seven of the 94 patients had a positive cTnT by test strip defined as >0.10 ng/ml. Twenty four patients died in the follow-up period (14 from cardiovascular causes). Twenty of the 24 non-survivors had an increased cTnT by test strip and 23 had increased cTnT by laboratory immunoassay. The outcome analysed by a Cox's proportional regression analysis showed that the factors which influenced survival significantly were cTnT, the presence of ischaemic heart disease, C reactive protein (CRP) and prealbumin. A logistic multivariate analysis revealed that age and CRP significantly influenced cTnT. A good correlation was found between cTnT determined by test strip and in the laboratory. CONCLUSION: cTnT is elevated in a large number of patients on regular haemodialysis and is a significant independent predictor of outcome. Increased cTnT is significantly predicted by age and CRP. PMID- 10462279 TI - Advanced glycated end-products (AGE) during haemodialysis treatment: discrepant results with different methodologies reflecting the heterogeneity of AGE compounds. AB - BACKGROUND: There has been much recent interest in accumulation of advanced glycation end-products (AGE) in uraemic patients. Analysis of AGE has been difficult, because commonly used methodologies, i.e. immunodetection assays or fluorescence measurements, reflect group reactivity and are not specific for chemically defined substances. Some investigators measured individual AGE compounds, e.g. pentosidine, carboxymethyllysine, pyrraline or imidazolone, but a systematic assessment of known compounds using specific HPLC methods in diabetic and non-diabetic end-stage renal disease (ESRD) patients during treatment has not been performed. METHODS: For the present study, the concentrations of early and late products of the Maillard reaction in plasma and ultrafiltrate were monitored during high-flux dialysis sessions in diabetic and non-diabetic patients. AGE were analysed by fluorescence spectroscopy and size exclusion chromatography with fluorescence detection. Specific HPLC methods were used to quantify the Amadori product fructoselysine and the AGE compounds pentosidine and pyrraline in acid or enzymatic hydrolysates. RESULTS: Using size exclusion chromatography, we confirmed a similar fluorescent peak distribution for diabetic and non-diabetic ESRD patients. Main fractions were found at approximately 70, approximately 14 and <2 kDa, confirming results obtained by other authors. In diabetic patients, the fluorescence intensity of the low molecular weight fraction was higher. Uraemic patients differed from controls mainly by the fluorescence of the low molecular weight fraction. The peak spectrum in ultrafiltrates was similar to that in plasma regarding low molecular weight fractions and the 14 kDa peak, but no protein-bound fluorescence was found at 70 kDa. HPLC analysis revealed a significant reduction of plasma pentosidine during high-flux dialysis in non diabetic (from 9.1+/-5.1 to 8.5+/-4.7 pmol/mg protein; P<0.05) and diabetic patients (from 10.0+/-9.1 to 6.8+/-4.0 pmol/mg protein; P<0.05). In contrast, plasma fructoselysine showed only a non-significant trend to decrease in diabetic (from 3.24+/-0.88 to 3.05+/-0.77 nmol/mg protein) and non-diabetic patients (from 2.69+/-0.52 to 2.56+/-0.50 nmol/mg protein). Pyrraline, a nonfluorescent late AGE product derived from reaction of 3-deoxyglucosone with lysine, could not be detected (detection limit approximately 40 pmol/mg protein). Comparing HPLC and size exclusion analysis, it was found that pentosidine accumulated in the range of low molecular weight substances and was removed by high-flux dialysis. CONCLUSIONS: High-flux dialysis reduces the plasma concentration of fluorescent AGE compounds, i.e. pentosidine, but the Amadori product fructoselysine is not removed, indicating that this compound is protein associated. PMID- 10462280 TI - Blood pressure variability as an adverse prognostic risk factor in end-stage renal disease. AB - BACKGROUND: Prospective and case-control studies show that blood-pressure variability is an independent risk factor for severe organ damage and cardiovascular events in hypertensives. We prospectively studied the association between systolic blood pressure variability and cardiovascular mortality and mortality from all causes in end-stage renal disease patients. METHODS AND RESULTS: The subjects were 144 patients (86 men, 58 women; mean age+/-SD, 52+/-13 years) who underwent dialysis in the same dialysis centre and were examined for blood-pressure variability. The study period was 38 months beginning in January 1995, during which six cardiovascular and seven noncardiovascular fatalities occurred. Coefficient of variation in systolic blood pressure in 1994, as an indicator of systolic blood pressure variability, ranged from 7.8 to 14.6%. Cumulative incidence of death from all causes was related to coefficient of variation in systolic blood pressure. The difference between the maximum and minimum systolic blood pressure (deltaSBP) in 1994 ranged from 44 to 146 mmHg (mean+/-SD, 78+/-13 mmHg) and correlated significantly with coefficient of variation in systolic blood pressure (r = 0.65, P<0.0001). Cox regression analysis was used to identify the independent predictors for mortality. The hazard ratio for death from all causes increased 1.63 times per 1% increase in coefficient of variation in systolic blood pressure (hazard ratio; 95% confidence interval: 1.63; 1.05-2.53) and 1.03 times per 1 mmHg increase in deltaSBP (1.08; 1.03-1.14). CONCLUSION: These results suggest that systolic blood pressure variability may be a significant prognostic factor in end-stage renal disease. PMID- 10462281 TI - The kidney during hibernation and arousal from hibernation. A natural model of organ preservation during cold ischaemia and reperfusion. AB - BACKGROUND: During hibernation the kidney is in a hypothermic condition where renal blood flow is minimal and urine production is much reduced. Periodical arousal from hibernation is associated with kidney reperfusion at increasing body temperature, and restored urine production rate. METHODS: To assess the degree of structural preservation during such extreme conditions, the kidney cortex was investigated by means of electron microscopy in the dormouse Muscardinus avellanarius during winter hibernation, arousal from hibernation and the summer active period. RESULTS: Results show that the fine structure of the kidney cortex is well preserved during hibernation. In the renal corpuscle, a sign of slight lesion was the focal presence of oedematous endothelial cells and/or podocytes. Proximal convoluted tubule cells showed fully preserved ultrastructure and polarity, and hypertrophic apical endocytic apparatus. Structural changes were associated with increased plasma electrolytes, creatinine and urea nitrogen, and proteinuria. During the process of arousal the fine structure of the kidney cortex was also well maintained. CONCLUSION: These results demonstrate that dormice are able to fully preserve kidney cortex structure under extreme conditions resembling e.g. severe ischaemia or hypothermic organ storage for transplantation, and reperfusion. Elucidation of the mechanisms involved in such a natural model of organ preservation could be relevant to human medicine. PMID- 10462282 TI - Rapid and accurate assessment of glomerular filtration rate in patients with renal transplants using serum cystatin C. AB - BACKGROUND: Assessment of renal function in patients with renal transplants is of great importance. Various studies have reported cystatin C as an easily and rapidly assessable marker that can be used for accurate information on renal function impairment. To date, no study is available to define the role of cystatin C in patients with renal transplants. METHODS: Thirty steady-state patients (50% male/50% female) with status post-kidney transplantation were studied. To assess renal function, cystatin C, creatinine clearance, serum creatinine, beta2-microglobulin (beta2M), and [125I]iothalamate clearance were determined. Correlations and non-parametric ROC curves for accuracy, using a cut off glomerular filtration rate (GFR) of 60 ml/min, were obtained for the different markers allowing for calculations of positive predictive values (PPV), positive likelihood ratios (PLR), specificity and sensitivity, respectively. Further, to evaluate the usefulness of these markers for monitoring, intraindividual coefficients of variation (CVs) for cystatin C and creatinine measurements were compared in 85 renal transplant patients. Measurements consisted of at least six pairs of results, which were obtained at different time points during routine follow-up. RESULTS: Cystatin C correlated best with GFR (r=0.83), whereas serum creatinine (r=0.67), creatinine clearance (r=0.57) and beta2M (r=0.58) all had lower correlation coefficients. The diagnostic accuracy of cystatin C was significantly better than serum creatinine (P=0.025), but did not differ significantly from creatinine clearance (P=0.76) and beta2M (P=0.43). At a cut-off of 1.64 mg/l, cystatin C has a PPV of 93%, PLR of 6.4, specificity 89% and sensitivity 70%, respectively. For beta2M, PPV 83%, PLR 1.7, specificity 67% and sensitivity 75% was seen at a cut-off of 3.57 mg/l. Accordingly, at a cut off of 125 micromol/l for serum creatinine, a PPV 76%, PLR 1.4, specificity 44% and sensitivity 80% was revealed. Finally, at a cut-off of 66 ml/min/1.73 m2 for creatinine clearance, the following characteristics were found: PPV 94%, PLR 7.7, specificity 89% and sensitivity 85%. The intraindividual variation of creatinine was significantly lower than that of cystatin C (P<0.001). With increasing concentrations, their ratios of CV tended towards a value of 1, demonstrating identical variability at low GFR. CONCLUSION: Together, our data show that in patients with renal transplants, cystatin C, in terms of PPV and PLR, has a similar diagnostic value as creatinine clearance. However, it is superior to serum determinations of creatinine and beta2M. The intraindividual variation of cystatin C is greater than that of creatinine. This might be due to the better ability of cystatin C to reflect temporary changes especially in mildly impaired GFR, most critical for early detection of rejection and other function impairment. Thus, cystatin C allows for rapid and accurate assessment of renal function (GFR) in renal transplants and is clearly superior to the commonly used serum creatinine. PMID- 10462283 TI - Simultaneous presentation of Goodpasture's disease and insulin-dependent diabetes mellitus. PMID- 10462284 TI - Unilateral renal cystic disease in adults. AB - Unilateral renal cystic disease (URCD) is morphologically indistinguishable from autosomal dominant polycystic kidney disease (ADPKD) except for its unilaterality. Unlike ADPKD, URCD patients show neither a genetic background nor progressive deterioration in renal function; thus, the differential diagnosis of URCD from ADPKD is important. Only a few cases of URCD have been reported. This study reports two cases of URCD in adults together with a literature review. We identified these two cases using abdominal computerized tomography and family screening with renal ultrasonography. PMID- 10462285 TI - De novo diabetes mellitus in kidney allografts: nodular sclerosis and diffuse glomerulosclerosis leading to graft failure. PMID- 10462286 TI - Steroid-sensitive nephrotic syndrome, sarcoidosis and thyroiditis--a new syndrome? PMID- 10462287 TI - Simultaneous acute renal and hepatic failure after ingesting raw carp gall bladder. PMID- 10462288 TI - Severe hypophosphataemia in paracetamol-induced oliguric renal failure. PMID- 10462289 TI - Successful prolonged use of an intracardiac catheter for dialysis. PMID- 10462290 TI - Trichosporon beigelii fungaemia in a patient with haemodialysis. PMID- 10462291 TI - Tuberculosis of the skull in a patient on maintenance haemodialysis. PMID- 10462292 TI - An unusual cause of post-biopsy oliguria in an allograft. PMID- 10462293 TI - Unexplained haematuria. PMID- 10462294 TI - Placement of an internal jugular dialysis catheter into the superior intercostal vein. PMID- 10462295 TI - Exercise induced macroscopic haematuria: run for a diagnosis? PMID- 10462296 TI - Ischaemic bowel disease in patients on continuous ambulatory peritoneal dialysis. PMID- 10462297 TI - The lady with a history of blood transfusion who developed palpable purpura and microhaematuria. PMID- 10462298 TI - Austria 19th century. An atlas on urinary sediment written by a surgeon and a chemist still of interest today. PMID- 10462299 TI - 'The spouse as a kidney donor: ethically sound?'. PMID- 10462300 TI - The Normal Haematocrit Trial in dialysis patients with cardiac disease. PMID- 10462301 TI - Haemodialysis for French diabetic patients. PMID- 10462302 TI - Acute renal failure in a young man, 9 days after a 5-h persisting erection following Viagra: is there a causal relationship? PMID- 10462303 TI - Temporal association between the ingestion of two consecutive doses of sildenafil 100 mg by a healthy subject and his presentation several days later with symptoms of acute renal failure which resolved without treatment. PMID- 10462304 TI - The effect of angiotensin II receptor antagonist on the exogenous erythropoietin requirement of haemodialysis patients. PMID- 10462305 TI - Another case of focal segmental glomerulosclerosis in an acutely uraemic patient following interferon therapy. PMID- 10462306 TI - Cefazolin serum concentrations with fixed intravenous dosing in patients on chronic hemodialysis treatment. PMID- 10462307 TI - The spectrum of acute renal failure in tumour lysis syndrome. PMID- 10462308 TI - Spousal renal donor transplants in India. PMID- 10462309 TI - Vasculitis look-alikes: variants of renal atheroembolic disease. PMID- 10462310 TI - Non-diabetic glomerulosclerosis in renal transplantation. PMID- 10462311 TI - Philadelphia chromosome (+) T-cell acute lymphoblastic leukaemia after renal transplantation. PMID- 10462312 TI - Anti-HCV seropositivity among haemodialysis patients of Iranian origin. PMID- 10462313 TI - Introduction: rising PSA after radical prostatectomy: a burning issue. PMID- 10462314 TI - The significance of recurrent PSA after radical prostatectomy: benign versus malignant sources. AB - The purpose of this article is to review the available means to investigate whether an elevated serum prostate-specific antigen (PSA) after radical prostatectomy may be explained by the presence of residual benign tissue. To answer this question, one may consider the following features: the kinetics of recurrent/ persistent PSA, the incidence of rising PSA in the presence of capsular incisions exposing benign glands only, the level of urinary PSA and the ratio of free/total PSA in the urine, the results of anastomotic biopsy samples, and the detection of circulating prostate cells by PSA reverse transcriptase polymerase chain reaction (RT-PCR) after surgery. Capsular incisions exposing benign tissue are not associated with a significant risk of biochemical failure. In case of an organ-confined cancer with negative surgical margins but a rising postoperative PSA, the systematic reevaluation of the initial pathological slides constantly shows capsular effraction or focal positive margins that have been overlooked at the first evaluation. Even when anastomotic biopsies document only benign tissue, the study of PSA doubling time is usually characteristic of the coexistence of residual tumoral cells. However, in a few cases, the persistent negative results of the detection of circulating prostate cells by PSA, RT-PCR in patients with organ-confined cancer and negative margins but elevated postoperative PSA might be explained by the presence of residual benign prostatic hyperplasia tissue. Most of the data in the literature are in favor of the responsibility of persistent/recurrent cancer in the recurring PSA rather than that of benign prostatic hyperplasia/normal residual tissue. Therefore, a persistent/recurrent detectable level of PSA is the serum after radical prostatectomy characterizes biochemical failure. PMID- 10462315 TI - How to explore the patient with a rising PSA after radical prostatectomy: defining local versus systemic failure. AB - We report on the various methods used to determine local or distant recurrences in patients with detectable serum prostate-specific antigen (PSA) after radical prostatectomy (RP). Studies show that variables that help predict distant metastases are PSA recurrence less than 2 years following surgery, tumors with Gleason score (GS) greater than 7, and positive seminal vesicles or positive lymph nodes at the time of surgery. In addition, studies in PSA kinetics show that short PSA doubling times, especially less than 6 months, are associated with distant recurrence and better correlated with the pattern and incidence of clinical recurrence than preoperative PSA, specimen GS, or stage alone. Studies show that although positive surgical margins are a significant risk factor for recurrence, only 40% to 50% of patients with positive margins developed an elevated PSA level within 5 years. When suspecting a local recurrence, transrectal ultrasound (TRUS) and TRUS-guided biopsies enhance the relatively inaccurate detection of local recurrence by digital rectal examination and initial prostate fossa biopsies. For distant recurrence, bone scintigrams of patients with a PSA recurrence following RP are only rarely positive and are found to have limited usefulness until the PSA increases to above 30 ng/mL. The role of immunoscintography to differentiate between local and distant recurrence is still evolving and requires further investigation. Further studies are clearly needed to enhance our ability to distinguish local from distant recurrence and to ultimately help guide therapy. PMID- 10462316 TI - Observation/delayed treatment for rising PSA after radical prostatectomy: pros and cons. AB - The protracted natural history of prostate cancer and the absence of a documented effective seconde line curative therapy, once primary treatment has failed, has led in the presence of biochemical failure after radical prostatectomy to a dual attitude: early treatment versus observation with delayed therapy. The objective of this review is to define the subsets of patients who might benefit from either of these attitudes. Depending on the risk of progression, three subgroups of patients may be individualized: a high-risk group (PN1, pT3 B, Gleason score equal or superior to 8), a moderate-risk group (pT3 A, NO with positive margins, and a Gleason score equal or less than 7), and a low-risk group (pT2 NO or pT3 A NO without positive margins and Gleason score equal or lower than 7). As of today, observation seems to be the appropriate option in men with a low or moderate risk of progression, whereas in the high-risk group, early therapy is a reasonable option. New treatment options with intermittent hormonal therapy or with combined adjuvant hormonoradiotherapy show a promising efficacy that may lead to reconsider this attitude. PMID- 10462317 TI - Radiation therapy for a rising PSA level after radical prostatectomy. AB - With the advent of routine prostate-specific antigen (PSA) testing in asymptomatic young men, the number of patients undergoing radical prostatectomy is increasing. Effective therapy for the 30% to 40% of men who recur following radical prostatectomy is essential. Salvage radiation therapy for a rising serum PSA level is an effective therapy for controlling local recurrence. The impact of salvage radiotherapy on disease-free survival is promising, but the guidelines of therapy are not well established. The best candidates for salvage radiotherapy are those with a serum PSA level < 2 ng/mL with no palpable recurrence and complete urinary continence. PMID- 10462318 TI - Hormonal manipulation for rising PSA after radical prostatectomy. AB - PSA recurrence after radical prostatectomy usually indicates recurrent prostate cancer. Identification of the recurrence site is difficult, but pathological and clinical features may suggest local versus distant recurrence. Radiographic techniques including transrectal ultrasonography, and 111indium capromab pendetide scans may help identify recurrences. The use of hormonal manipulation for rising PSA after radical prostatectomy is controversial. Androgen deprivation has been a mainstay of the management for advanced prostate cancer. The timing of such therapy is debatable, and early therapy in an asymptomatic patient may not correlate with improved survival. Maximal androgenic blockade with castration and nonsteroidal antiandrogens may offer a modest survival benefit in selected patients. Novel potency-sparing therapies with antiandrogens and finasteride afford an improved patient lifestyle, with questionable effects on survival. Intermittent androgen suppression is an experimental treatment modality that may reduce the side effects of castration. Ongoing studies are being performed to clarify these controversies, and the variety of treatment options allows patients great flexibility in considering quality of life and effective cancer control. PMID- 10462319 TI - The role of dietary manipulation in biochemical recurrence of prostate cancer after radical prostatectomy. AB - In 1999, we continue to be faced with difficulties in treating prostate cancer, which remains the leading cause of cancer and the second leading cause of cancer related deaths in men. In the prostate-specific antigen (PSA) era, we must also determine which therapies, if any, are appropriate for the treatment of a biochemical or PSA relapse. In view of the limited number of efficacious and durable treatments for prostate cancer recurrence, the medical community has had to investigate nontraditional therapies. In the recent past, an expanding body of evidence has implicated certain nutrients in carcinogenesis and cancer progression. This article discusses the role of diet in prostate carcinogenesis and the rationale for dietary manipulation as a treatment strategy for the prevention of primary and secondary (recurrent) prostate cancer. PMID- 10462320 TI - Rising PSAs after primary therapy: active or passive intervention. AB - Prostate cancer that has relapsed biochemically after primary therapies, such as prostatectomy or radiation, remains a therapeutic challenge in that no standard treatment option exists for this patient. These patients are often young and may be offered androgen ablation as the mainstay of treatment. Many patients do not wish to undergo a regimen that may be associated with a variety of side effects that will impact on their quality of life. Delaying hormonal treatment in this group does not compromise survival and patients may try a variety of approaches in an attempt to control rising PSAs. Therefore, these patients are an interesting subgroup for whom immunological and alternative therapies may prove to be beneficial. We review new approaches for this population of men, which result in antitumor effects with minimal toxicities. PMID- 10462321 TI - Introduction: Hodgkin's lymphoma--pathology, pathogenesis, and treatment. PMID- 10462322 TI - Hodgkin's lymphomas: classification, diagnosis, and grading. AB - Much has been learned in the past decade from the study of tissues involved by the disorder long known as Hodgkin's disease. Two important discoveries have prompted changes in classification and nomenclature: first, the recognition that there are two distinct lymphomas encompassed within this category (classical and nodular lymphocyte-predominant [NLP] types), and second, the discovery that the Reed-Sternberg (RS) cells in most cases are monoclonal B cells. Thus "Hodgkin's disease" comprises two distinct lymphomas, deserving of a name change, to "Hodgkin's lymphomas" (HLs). The immunophenotype and genetic features of both classical HL and NLPHL have been defined. These are useful in the subclassification of HL and in distinguishing HL from two recently described, aggressive lymphomas that were in the past often diagnosed as HL: anaplastic large-cell lymphoma, T-cell type (ALCL), and T-cell/histiocyte-rich large B-cell lymphoma (T/HRBCL). Despite the advances of recent years, many questions remain to be answered, and these will provide the challenges of the next decade. PMID- 10462323 TI - Cellular origin and clonality of classic Hodgkin's lymphoma: immunophenotypic and molecular studies. AB - The cellular origin of Hodgkin and Reed-Sternberg (HRS) cells, the neoplastic cells of classic Hodgkin's lymphoma (HL), resisted clarification until the second half of this decade. One major obstacle to successful experimental investigations was the rarity of the HRS cells in the tissue affected by HL. Immunophenotypical studies using monoclonal antibodies already pointed in the early 1980s towards a lymphocytic origin for HRS cells, but were not definitive because of the usually variable expression of B-cell and/or T-cell antigens, and the additional expression of markers typical for other cell lineages, especially dendritic cells. Attempts to elucidate the cellular derivation of HRS cells by demonstrating the clonal rearrangements of the immunoglobulin (Ig) or T-cell receptor (TCR) genes in the DNA of whole-tissue extracts also remained inconclusive due to inconsistent results. In frustration with whole-tissue DNA studies, genetic approaches were turned to the single cell level. Among the techniques developed for the isolation of HRS cells, the extraction of single CD30+ HRS cells from immunostained frozen sections by means of hydraulic micromanipulation proved to be the most suitable. Using this method, monoclonal Ig gene rearrangements were detected in the HRS cells in 36 of 38 (95%) HL cases. Sequence analysis demonstrated high loads of somatic mutation in the rearranged variable regions. Molecular investigation of three cases of HL occurring in association with non-Hodgkin's lymphomas (NHLs) showed that all of the lymphoma lesions had an identical precursor with the molecular features of a germinal center B cell. In summary, these findings indicate that (1) approximately 95% of classic HLs originate from B cells; (2) the direct cellular precursors of the HRS cells are germinal center B cells; (3) the transforming event that causes HL leads to the complete morphologic and immunophenotypical change of the HRS cell precursors; and (4) the HRS cell population of a given case exclusively arises from a single transformed cell and expands by clonal growth. It remains to be shown whether the 5% of HLs for which a B-cell derivation could not be demonstrated are T-cell related. PMID- 10462324 TI - Cellular origin of nodular lymphocyte-predominant Hodgkin's lymphoma: immunophenotypic and molecular studies. AB - The lymphocyte-predominant (LP) type of Hodgkin's lymphoma (HL) is a unique subtype with characteristic tumor cells (lymphocytic and/or histiocytic [L&H] cells) in associated macronodular structures that resemble progressively transformed germinal centers (PTGCs). Immunohistochemical studies have provided strong evidence that L&H cells are of B-cell lineage and recent molecular studies suggested they are transformed centroblasts. A major clonal population is detectable at presentation, with the immunoglobulin heavy-chain gene often showing evidence of continued somatic hypermutation. In developed nations, Epstein-Barr virus (EBV) is infrequently associated with L&H cells and is probably not involved in the pathogenesis of this disease. L&H cells are frequently surrounded by CD3+, CD4+, CD57+, and CD40L- T cells, but the significance of this T-cell rosetting is unclear. LPHL may be associated with concurrent or subsequent large B-cell lymphoma, and there is evidence of a clonal relationship between the two entities. LPHL may also have nodules or large areas that resemble histiocyte-rich B-cell lymphoma (HRBCL). It is likely that at least some cases of HRBCL arise from LPHL. The same may be true of T-cell-rich B-cell lymphoma. Little is known about cytogenetic abnormalities, the cytokine expression profile, and the expression of several functionally important molecules that have been demonstrated in the Reed-Sternberg (RS) cells of classical HL. The challenge for the future is to obtain a more comprehensive molecular profile of L&H cells and their associated T lymphocytes, so as to provide a framework for eventual elucidation of the pathogenesis of this type of HL. PMID- 10462325 TI - Immune reactions in classical Hodgkin's lymphoma. AB - The immune reaction in classical Hodgkin's lymphoma (HL) can be separated into an inflammatory response in the involved tissues and a generalized immune response in the patient. The local immune reaction in HL is by far the most prominent among all tumors, with the exception of so called T-cell-rich B-cell lymphoma, a subtype of large-cell B-cell lymphoma. The general immune response in patients with HL is best described as an acquired cellular immune deficiency, most likely a result of the presence of tumor, although some data in the literature suggest a preexisting immune deficiency. The cellular origin of Reed-Sternberg (RS) cells in classical and nodular lymphocyte-predominant HL is discussed elsewhere. RS cells and their mononuclear variants can be considered as clones of neoplastic B cells that, by secreting potent cytokines/chemokines, not only cause the symptoms of HL but also promote their own growth and evade immune surveillance. The characteristic histologic features of HL--the prominent inflammatory response, the influx of eosinophils, and the presence of fibrosis and sclerosis--may also result from the expression of a range of surface molecules and the production of cytokines and chemokines by RS cells. PMID- 10462326 TI - Epstein-Barr virus and other candidate viruses in the pathogenesis of Hodgkin's disease. AB - The epidemiologic and clinicopathologic features of Hodgkin's disease (HD) suggest that an infectious agent is involved in the etiology. Over the last 12 years, evidence has accumulated suggesting that Epstein-Barr virus (EBV) is associated with a proportion of cases: EBV genomes are present in Reed-Sternberg (HRS) cells and viral proteins including LMP1, which has oncogenic potential, are expressed. HD has a complex epidemiology and EBV-associated cases are not randomly distributed. Disease occurring in early childhood and older adult age groups is more likely to be EBV-associated than for young adult cases. Paradoxically, there is more evidence supporting an infectious etiology in the latter group of younger patients. Defective EBV genomes and "hit and run" mechanisms involving EBV cannot account for all cases, and the direct involvement of known viral agents, including other lymphotropic herpesviruses, has largely been excluded. Hitherto unknown virus may be responsible for the peak incidence in young adults, which is a feature of HD in developed countries. PMID- 10462327 TI - Treatment of early-stage Hodgkin's disease. AB - The management of early-stage Hodgkin's disease has been evolving over the past 25 years, largely due to recognition of early and late complications associated with radiation therapy and the demonstration of minimally toxic but effective chemotherapy. Thus, extended-field radiation is no longer the "gold standard" of treatment. Rather, combined modality approaches with abbreviated chemotherapy and limited radiation, which obviates the need for precise staging offered by laparotomy, provide excellent results with the potential for fewer adverse late effects. Several controlled clinical trials are ongoing to determine the minimal duration of chemotherapy and extent and dose of radiotherapy that will not compromise the excellent cure rate of early-stage disease. PMID- 10462328 TI - Treatment of advanced Hodgkin's lymphoma: standard and experimental approaches. AB - The introduction of polychemotherapy and improved radiation techniques has transformed Hodgkin's lymphoma from an incurable disease to a malignancy with one of the highest cure rates. Milestones were the development of the MOPP (mechlorethamine, vincristine, procarbazine, and prednisone) and ABVD (doxorubicin, bleomycin, vinblastine, and dacarbazine) regimens. Radiotherapy is commonly used, although its precise role has not been defined for patients with advanced-stage disease. More recently, dose-intensified schedules such as Stanford V (doxorubicin, vinblastine, mechlorethamine, vincristine, bleomycin, etoposide, and prednisone) were shown to be effective in this group of patients. In particular, the BEACOPP regimen (bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone), in both standard and escalated doses, has produced impressive results in a randomized three-arm study when compared with COPP (cyclophosphamide, vincristine, procarbazine, and prednisone)/ABVD. The significantly higher rates of complete remission (CR) and freedom from treatment failure (FFTF) suggest that the new BEACOPP regimen improves efficacy, but definitive conclusions require further years of follow-up evaluation. Interestingly, BEACOPP abrogates the impact of the newly described seven-factor prognostic scoring system that was reported for patients treated with MOPP/ABVD or similar regimens. The prognostic index includes factors such as serum albumin, hemoglobin, male sex, stage IV disease, age more than 45 years, white blood cell count, and lymphocyte count. Whereas patients with Hodgkin's lymphoma have a good prognosis on first diagnosis, those with relapsed or refractory disease face a poor outcome. PMID- 10462329 TI - Radiation therapy in Hodgkin's lymphoma. AB - Radiation therapy continues to play a primary role for the definitive treatment of early-stage Hodgkin's disease. Its value as an adjunct for chemotherapy in patients with unfavorable features such as bulky disease is also obvious. The indications for radiation therapy in addition to chemotherapy for patients with advanced-stage disease are less clear. Nevertheless, there is adequate evidence that consolidative irradiation is of benefit in at least a subset of such patients. Future directions in radiation therapy for Hodgkin's disease include efforts to reduce treatment extent (both field size and dose) without compromising disease control; further technological advances in improving planning and treatment equipment, to allow better tumor targeting while minimizing dose to surrounding normal tissue; close follow-up evaluation of patients treated for Hodgkin's disease, which may allow better selection of patients for the different treatment schemes and tailoring therapy according to risks for relapse as well as treatment complications; optimizing a follow-up plan; and implementing preventive measures for long-term complications of treatment. The goal is not only to maximize survival but also to optimize patient quality of life. PMID- 10462330 TI - High-dose therapy with hematopoietic transplantation for Hodgkin's lymphoma. AB - The use of combination chemotherapy has been successful in treating advanced or widespread Hodgkin's disease (HD). For patients with relapsed disease, the long term prognosis is much poorer, despite the good rate of reinduction into a second or further remission using conventional treatments. The majority of such patients will ultimately relapse again if given only conventional salvage chemotherapy. The transplantation or reinfusion of hematopoetic stem cells allows the use of higher doses of cytotoxic drugs that would normally be precluded by myelotoxicity. Hematopoiesis rescue by autologous transplantation of stem cells has become an accepted form of treatment for HD that has failed to respond to first-line chemotherapy, which has relapsed within 1 year after chemotherapy, or which has relapsed on more than one occasion. Dose escalation with autologous stem cells is fashionable, but there is an increasing tendency to consider allogeneic transplantation in HD. There may be some limited graft-versus Hodgkin's lymphoma effect, but this is outweighed by the greatly increased treatment toxicity associated with the allogeneic procedure. It is possible, however, that modern low-intensity conditioning regimens, the so-called mini allograft approach, may increase the use of allogeneic transplantation for poor prognosis Hodgkin's lymphoma patients in the future. PMID- 10462331 TI - Treatment of pediatric Hodgkin's lymphoma. AB - Today the majority of children and adolescents diagnosed with Hodgkin's lymphoma will enjoy long-term disease-free survival. As a result, contemporary treatment strategies have focused on reducing therapy for patients with favorable disease presentations and reserved aggressive treatment modalities for patients with relapsed or refractory disease. The desire to avoid late treatment toxicity has prompted refinements in therapy designed to reduce growth impairment, second malignancy, and life-threatening organ dysfunction in long-term survivors. Treatment with radiation therapy alone is recommended only for older patients with localized disease who have achieved skeletal maturity, but requires surgical staging and places greater volumes of normal tissues at risk for late carcinogenesis. Treatment with chemotherapy alone avoids the long-term growth, organ dysfunction, and solid tumor induction associated with high-dose, extended field radiation. However, these protocols prescribe higher cumulative doses of alkylating agent chemotherapy, which may increase the risk of treatment complications from myelosuppression, gonadal injury, and secondary leukemia. Combined modality therapy regimens have resulted in excellent treatment outcomes and reduced the incidence of treatment sequelae by utilizing lower doses and smaller volumes of radiation therapy and fewer cycles of less toxic chemotherapy in clinically staged children. Risk-adapted therapies using two to four cycles of multiagent chemotherapy and lower radiation doses and volumes have maintained excellent disease-free survival rates in clinically staged patients with localized favorable disease presentations. Novel approaches including compacted dose-intensive multiagent chemotherapy are currently under investigation with the objectives of improving outcome and reducing treatment sequelae in patients with advanced and unfavorable disease. PMID- 10462332 TI - Expanded role of group A streptococci in children with upper respiratory infections. PMID- 10462333 TI - Comparison of procalcitonin with interleukin 8, C-reactive protein and differential white blood cell count for the early diagnosis of bacterial infections in newborn infants. AB - OBJECTIVE: To evaluate procalcitonin (PCT) as a test for early diagnosis of bacterial infections (BI) in newborn infants and to compare the results of PCT with those of interleukin 8 (IL-8), C-reactive protein (CRP) and differential white blood cell count. STUDY DESIGN: PCT was prospectively measured along with IL-8, CRP and differential white blood cell counts and blood cultures in 197 newborn infants at the first suspicion of bacterial infection. PCT, IL-8, CRP and differential white blood cell counts were analyzed for sensitivity, specificity and positive and negative predictive values after receiver operating characteristic curve analysis for best thresholds. The kinetics of PCT was determined in infants with and without BI. RESULTS: Forty-six infants were diagnosed clinically as having BI, of whom 9 had BI with positive blood cultures. At a cutoff value of 0.50 microg/l, PCT detected combined culture-proved and clinical BI with a sensitivity of 57% (95% confidence interval, 41%, 71%) and a specificity of 66% (95% confidence interval, 57%, 74%). The combination of IL-8 > or =70 ng/l and/or CRP >10 mg/l achieved a sensitivity of 91% (95% confidence interval, 79%, 98%) and a specificity of 73% (95% confidence interval, 64%, 81%). PCT values of infected and not infected infants tended to rise for 24 h after initial evaluation and then decreased. CONCLUSION: The combination of IL-8 and CRP is more reliable than PCT as a test for early diagnosis of BI in newborn infants. PMID- 10462334 TI - An open randomized controlled trial of infection prevention in child day-care centers. AB - OBJECTIVE: Child care outside the home increases children's infections substantially. We have to evaluate the possibilities for reducing the transmission of infections by an infection prevention program. DESIGN AND METHODS: A 15-month randomized controlled trial involving 20 day-care centers was conducted to evaluate the efficacy of an infection prevention program. The program was introduced in 10 centers and the other matched 10 centers served as controls. Records were made of the occurrence of infections and absences from care or work because of infections among the children, their parents and the personnel of the day-care centers. RESULTS: Both the children and the personnel in the program centers had significantly fewer infections than those in the control centers, the reduction being 9% [95% confidence interval (95% CI), 4 to 16%, P < 0.002] among 3-year-old children and 8% (95% CI 0 to 14%, P = 0.049) among the older children. The children at the program centers received 24% fewer prescriptions of antimicrobials (95% CI 22 to 27%, P < 0.001). Likewise there were 2.5 man-year fewer absences from work on the part of parents because of a child's illness during 1 year in the program centers, a 24% difference (95% CI 18 to 29%, P < 0.001). CONCLUSIONS: Effective prevention of infections is possible in child day-care centers, and this can benefit both the families and the personnel. Such a program is cost-effective even if a specialist nurse implements it. PMID- 10462335 TI - Impact of routine infant and adolescent hepatitis B vaccination in Tuscany, Central Italy. AB - BACKGROUND: In Italy routine infant and adolescent immunization against hepatitis B was introduced in 1991. OBJECTIVE: Evaluation of (1) coverage with three doses of hepatitis B vaccine in infants and adolescents; (2) seroconversion to anti hepatitis B surface antigen antibody (anti-HBs) in adolescents receiving three doses of vaccine; (3) concordance of coverage rates in infants with prevalence of neutralizing antibodies in sera from anonymous children; (4) trend of notified cases of acute hepatitis B. METHODS: A sample of infants and adolescents living in Tuscany was studied during 6 years (1992 through 1997) by matching birth records and immunization certificates. Sera from 139 adolescents who completed the vaccination course and from 159 anonymous children belonging to immunized cohorts (1 to 5 years) were tested with a quantitative anti-HBs assay. Incidence of acute hepatitis B by age was calculated from regional statistics on notified infectious diseases between 1992 and 1996. RESULTS: Overall 10,606/11,164 (95%) infants and 10,599/11,100 (95%) adolescents received 3 doses of vaccine. Seroconversion to anti-HBs was detected in 98% of adolescent vaccinees. Anti-HBs titers > or =10 IU/l were detected in 87% of children. A 49% decline of acute hepatitis B cases was registered between 1992 and 1996 in 15- to 24-year-olds living in Tuscany. No case occurred in vaccinated adolescents. CONCLUSIONS: Coverage against hepatitis B is excellent in cohorts subject to mandatory immunization. If efforts to vaccinate are maintained at these levels, elimination of hepatitis B virus transmission could occur within few decades in Italy. PMID- 10462336 TI - Efficacy of and adherence to highly active antiretroviral therapy in children infected with human immunodeficiency virus type 1. AB - BACKGROUND: Clinical trials in adults have demonstrated the efficacy of highly active antiretroviral therapy (HAART) to suppress replication of HIV-1 to nondetectable levels, but lower success rates have been observed in practice. We sought to determine the efficacy of HAART in our population of HIV-1-infected children and to identify determinants of efficacy, especially the role of adherence to prescribed antiretrovirals. METHODS: The viral load and CD4+ T cell responses of 72 children with perinatally acquired HIV-1 treated with HAART including a protease inhibitor for at least 90 days were examined retrospectively in relation to adherence, as measured by pharmacy records for the first 180 days of HAART. RESULTS: Patients were defined as adherent if > or =75% of protease inhibitors and > or =75% of all antiretroviral prescriptions were filled. Of the 42 patients (58%) who were adherent, nondetectable viral loads were achieved and maintained in 22 (52%). A Kaplan-Meier plot showed a drop-off in patients maintaining a nondetectable viral load after 200 days. Higher initial viral load was the only pretreatment factor that identified adherent patients at risk for treatment failure. Only 3 (10%) nonadherent patients maintained a viral load of <400 copies/ml. The adherent group had a prompt and sustained increase in CD4+ T cell counts. CONCLUSIONS: HAART can achieve control of viral replication in HIV-1 infected children who adhere to therapy. However, treatment failure is likely unless there is a high level of adherence. Nonadherence to therapy is common and might be the major impediment to successful treatment of children infected with HIV-1. PMID- 10462337 TI - Mortality after the first year of life among human immunodeficiency virus type 1 infected and uninfected children. AB - BACKGROUND: HIV-infected and uninfected children who survived their first year of life were prospectively followed in Malawi to assess levels of mortality and related risk factors during the second and third years of life. METHODS: Children with known HIV status from an earlier perinatal intervention trial were enrolled. These children [HIV-infected (Group A); HIV-uninfected but born to HIV seropositive mothers (Group B); and children born to HIV-seronegative mothers (Group C)] were followed every 3 months until age 36 months. Mortality data were collected at each visit. Immunologic data (CD4+ percent) were collected at or immediately after enrollment. RESULTS: Overall 702 children were enrolled and 83 children died during follow-up. The mortality rate per 1000 person years of observation was 339.3 among Group A children, 46.3 among Group B children and 35.7 among Group C children. Among HIV-infected children the cumulative proportion surviving to age 24 months was 70% and those surviving to age 36 months was 55%. By age 32 months none of the severely immunosuppressed (CD4% < 15%) children had survived. The mortality differentials between HIV-infected and uninfected children persisted after adjusting for several risk factors. The major causes of death among infected children (n = 52) were wasting and respiratory conditions. CONCLUSIONS: Although all HIV-infected children had received childhood immunizations, mortality was high. Management of these children should include aggressive antimicrobial treatment, and evaluation of prophylactic regimens should be considered. PMID- 10462338 TI - Retreatment with higher dose interferon alpha in children with chronic hepatitis B infection. AB - BACKGROUND: More than 50% of children with chronic hepatitis B infection do not respond to interferon-alpha (IFN-alpha) treatment and are prone to have progressive liver disease. The best treatment modality is unknown in these children. The aim of this study was to evaluate the possible benefit of a second higher dose IFN-alpha therapy for children with chronic hepatitis B diseases who failed previous therapy. METHODS: Twenty-four children with chronic hepatitis B infection who had not responded to previous IFN-alpha treatment were enrolled into the study. All were hepatitis B virus DNA- and hepatitis B e antigen positive for >6 months after initial treatment. They received 10 megaunits (MU)/m2 of IFN-alpha 2a three times a week for 24 weeks. Liver function tests, hepatitis B virus markers and hepatitis B virus DNA were determined regularly during treatment and follow-up. A complete response was defined as clearance of both hepatitis B virus DNA and hepatitis B e antigen (HBeAg). RESULTS: At the end of therapy 8 (33.3%) patients cleared hepatitis B virus DNA and seroconverted to anti-HBeAg. Patients were followed for an average period of 12.2 +/- 4.7 months after retreatment. During follow-up an additional 4 patients cleared hepatitis B virus DNA and seroconverted to anti-HBe, whereas one seroconverted patient became HBeAg-positive again. Thus 11 patients (45.8%) had complete response at the end of the follow-up period. Alanine aminotransferase normalized in 11 responder patients and in 5 nonresponders. Positive predictive factors were low baseline titers of hepatitis B virus DNA and elevated transaminase values (> 100 IU/l). CONCLUSIONS: IFN-alpha retreatment with a higher dose may be an alternative modality for treatment of children with chronic hepatitis B infections who failed previous IFN-alpha, especially in those with favorable predictive factors. PMID- 10462339 TI - Epstein-Barr virus infection in sixty pediatric liver graft recipients: diagnosis of primary infection and virologic follow-up. AB - BACKGROUND: Epstein-Barr virus (EBV) is an important cause of infection after pediatric liver transplantation. Earlier detection of EBV could result in shortening the delay in diagnosis and allow better management of a pediatric high risk population. OBJECTIVES: To determine the timing of EBV primary infection after graft and to compare the performances of different assays for an early detection of the virus. METHODS: Sixty pediatric liver graft recipients were followed. Kinetics of appearance of different EBV serologic parameters (anti-EBV IgG, -IgM and -IgA, and anti-EBV nuclear antigen-IgG) and of the viral DNA in peripheral blood lymphocytes by PCR were compared. RESULTS: Thirty-six patients had a primary EBV infection. The first PCR and IgM positive result appeared after a mean delay of 56 and 61 days, respectively, and preceded the IgG response (mean delay, 143 days). Most of the studied patients (13 of 16) developed anti-EBV-IgA and only 3 developed anti-EBNA-IgG during the follow-up period. CONCLUSIONS: EBV primary infection occurred in most cases during the first 2 months after graft. The IgG response was delayed. The best performance was obtained by PCR. However, the IgM test compared well with the PCR and could be a more widely accessible measure to follow regularly. PMID- 10462340 TI - Amphotericin B lipid complex in pediatric patients with invasive fungal infections. AB - BACKGROUND: Lipid formulations of amphotericin B have been recently introduced for treatment of invasive fungal infections. However, little is known about their role in pediatric populations. METHODS: We studied the safety and antifungal efficacy of amphotericin B lipid complex (ABLC, Abelcet) in 111 treatment episodes in pediatric patients through an open label, emergency use multicenter study. Patients with invasive fungal infections were enrolled if they had mycoses refractory to conventional antifungal therapy, if they were intolerant of previous systemic antifungal agents or concomitant nephrotoxic drugs or if they had preexisting renal disease. RESULTS: All 111 treatment episodes were evaluable for safety and 54 were evaluable for efficacy. The mean serum creatinine for the study population did not significantly change between baseline (1.23 +/- 0.11 mg/dl) and cessation of ABLC therapy (1.32 +/- 0.12 mg/dl) during 6 weeks. There were no significant differences observed between initial and end-of-therapy levels of serum potassium, magnesium, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and hemoglobin. However, there was an increase in mean total bilirubin (3.66 +/- 0.73 to 5.31 +/- 1.09 mg/dl) at the end of therapy (P = 0.054). Among 54 cases fulfilling criteria for evaluation of antifungal efficacy, a complete or partial therapeutic response was obtained in 38 patients (70%) after ABLC therapy. Complete or partial therapeutic response was documented in 56% of cases with aspergillosis (n = 25) and in 81% (n = 27) with candidiasis. Among premature infants (n = 8) and allogeneic marrow recipients (n = 14), response rates were 88 and 57%, respectively. Response was similar in those patients enrolled because of intolerance to previous antifungal therapy or because of progressive infection. CONCLUSIONS: These data support the use of ABLC for treatment of invasive fungal infections in pediatric patients who are intolerant of or refractory to conventional antifungal therapy. PMID- 10462341 TI - Verocytotoxin-producing Escherichia coli infection in household members of children with hemolytic-uremic syndrome in The Netherlands. AB - BACKGROUND: Strains of verocytotoxin-producing Escherichia coli (VTEC) belonging to serogroup O157 (O157 VTEC) can cause a spectrum of disease that includes nonspecific diarrhea, hemorrhagic colitis and the diarrhea-associated form of the hemolytic uremic syndrome (D+ HUS). METHODS: We conducted a retrospective study of 34 children with D+ HUS caused by O157 VTEC to determine the frequency of VTEC infection in their household members. RESULTS: Gastrointestinal tract symptoms were reported in 1 or more household contacts of 17 (50%) of the 34 index cases. Of the 26 household members with gastrointestinal tract symptoms, 15 were parents and 11 were siblings. Evidence of VTEC infection was reported in 1 or more household contacts in 23 (68%) of the 34 families (in 46% of the siblings and in 28% of the parents). Nineteen (48%) siblings had a positive stool sample and in only 5 (12%) of the siblings IgM class serum antibodies to O157 lipopolysaccharide (LPS) were detected. Nineteen (31%) parents had a positive stool sample. Antibodies to O157-LPS were not detected in any of the parents. The occurrence of (bloody) diarrhea significantly correlated with the occurrence of IgM class serum antibodies to O157-LPS. CONCLUSIONS: It was concluded that household members of children with D+ HUS are often asymptomatically infected with O157 VTEC. Differences in the pathogenesis of the infection between infected individuals may be related to differences in the number of ingested O157 VTEC bacteria and to differences in susceptibility. PMID- 10462342 TI - Etiology of acute lower respiratory tract infection in Central Australian Aboriginal children. AB - BACKGROUND: Aboriginal children in central Australia have attack rates for acute lower respiratory tract infection (ALRI) that are similar to those in developing countries. Although mortality rates are much lower than in developing countries, morbidity is high and ALRI is still the leading cause of hospitalization. However, there are no data on the etiology of ALRI in this population. METHODS: We prospectively studied 322 cases of ALRI in 280 Aboriginal children admitted to the hospital. Blood, urine and nasopharyngeal aspirate samples were examined for evidence of bacterial, viral and chlamydial infection. RESULTS: The combination of blood culture, viral studies and chlamydial serology provided at least 1 etiologic agent in 170 of 322 (52.5%) cases. Assays for pneumolysin immune complex and pneumolysin antibody increased etiologic diagnosis to 219 (68.0%). Blood cultures were positive in 6% but pneumolysin immune complex and pneumolysin antibody studies were positive in one-third of cases. Evidence of viral infection was present in 155 (48%) of cases compared with 12% in controls (P < 001). There were only 7 possible cases and 2 definite cases of Chlamydia trachomatis and 3 cases of Chlamydia pneumoniae. Coinfection was common in these children. CONCLUSION: These findings have implications for both standard treatment protocols and vaccine strategies. The high rate of coinfection may make it difficult to develop simple clinical predictors of bacterial infection. In the setting of a developed country with efficient patient evacuation services, management algorithms that focus on disease severity and need for hospital referral will be most useful to health staff in remote communities. Pneumococcal conjugate vaccines will be required to reduce the high attack rate of pneumococcal disease. PMID- 10462343 TI - Anti-retroviral therapy for HIV-infected children--toward maximal effectiveness. AB - Viral suppression may be more difficult to achieve in children compared to adults. However, appropriate regimens at adequate dosages combined with monitoring of adherence and virologic response should allow children with HIV infection to realize a sustained benefit from combination anti-retroviral therapy. PMID- 10462344 TI - NSAIDS and necrotizing fasciitis. PMID- 10462345 TI - Pleural empyema. PMID- 10462346 TI - Cutaneous phaeohyphomycosis caused by Curvularia lunata and a review of Curvularia infections in pediatrics. PMID- 10462347 TI - A sixteen-year-old male youth with fever, dysphagia and weight loss. PMID- 10462348 TI - Most cerebrospinal fluid cultures in children with bacterial meningitis are positive within two days. PMID- 10462349 TI - Amantadine therapy for influenza type A-associated encephalopathy. PMID- 10462350 TI - Bacteremic pneumonia caused by penicillin-resistant Streptococcus pneumoniae in siblings. PMID- 10462351 TI - Persistent candidemia in a premature infant treated with fluconazole. PMID- 10462352 TI - Oerskovia xanthineolytica peritonitis: case report and review. PMID- 10462353 TI - Invasive aspergillosis in two patients with Pearson syndrome. PMID- 10462354 TI - Spectral gradient acoustic reflectometry: detection of middle ear effusion in suppurative acute otitis media. PMID- 10462355 TI - Accidental overdose of vancomycin in preterm twins. PMID- 10462356 TI - Posttraumatic invasive Aspergillus fumigatus wound infection. PMID- 10462357 TI - Sex-specific differences in serum vitamin A values after measles immunization. PMID- 10462358 TI - Clinical impact of upper gastrointestinal tract injuries in glyphosate-surfactant oral intoxication. AB - Fifty patients with glyphosate-surfactant oral ingestion were studied with upper gastrointestinal (UGI) endoscopic grading using Zargar's modified grading system for mucosal corrosive injury. Esophageal injury was seen in 68% of the patients, gastric injury in 72%, and duodenal injury in 16%. There were no grade 3 injuries. The upper gastrointestinal tract injuries caused by glyphosate surfactant were minor in comparison with those by other strong acids. The WBC count, amount of glyphosate-surfactant ingested, length of hospital stay and the occurrence of serious complications increased markedly in the group which had grade 2 esophageal injuries. Thus, the severity of the esophageal injuries may be a prognostic factor for the patient with glyphosate-surfactant ingestion. The UGI endoscopy may be indicated for grading esophageal injury in patients who have ingested glyphosate-surfactant in amounts greater than 100 ml. Physicians should pay more attention to the patients with grade 2 or 3 esophageal injuries to prevent serious complications and to provide aggressive supportive care. PMID- 10462359 TI - Human ocular effects from self-reported exposures to Roundup herbicides. AB - We evaluated ocular effects from reported human exposures to Roundup herbicides based on 1513 calls to an American Association of Poison Control Centers (AAPCC) certified regional poison center during the years 1993 through 1997. The preponderance of reported exposures were judged by poison center specialists to result in either no injury (21%) or transient minor symptoms (70%). There was some temporary injury in 2% of cases; one injury took more than 2 weeks to resolve. In no instance did exposure result in permanent change to the structure or function of the eye. Since the representativeness of calls to poison control centers is unknown, several interpretations of these findings are possible. The most conservative interpretation is that there were no serious ocular effects from exposure to Roundup herbicides during a 5 year period among callers to a single regional poison center. A less conservative interpretation would be that severe ocular effects from Roundup exposures are rare among users of these products. PMID- 10462360 TI - Inhibitory effects of H2-receptor antagonists on platelet function in vitro. AB - To evaluate in vitro inhibitory effects of four types of histamine H2-receptor antagonist (H2-receptor antagonists), famotidine, roxatidine, cimetidine and ranitidine, on platelet function, we examined aggregating potency and P-selectin levels with agonist-induced aggregation. Ranitidine and cimetidine inhibited, in concentration of 0.35 mM, the secondary aggregation induced by 5 microM adenosine diphosphate (ADP), the aggregation induced by 1 microg/mL collagen and 3 microM arachidonic acid. All of H2-receptor antagonists inhibited, in concentration of 1.4 mM, the aggregation induced by ADP, collagen and arachidonic acid. Ranitidine and cimetidine reduced markedly, in same concentration, P-selectin levels after induction of aggregation by 5 microm ADP, 1 microg/mL collagen and 3 microM arachidonic acid. When classified by the strength of inhibitory action, ranitidine and cimetidine were strong, followed by famotidine and roxatidine. It is considered that inhibitory effects of H2-receptor antagonists on platelet function are weaker than those of acetylsalicylic acid (ASA), since ASA inhibited platelet aggregation in concentration of 100 microM. No relationship was observed between inhibitory effects of H2-receptor antagonists on platelet aggregation induced by above agonists and the presence or absence of imidazole ring in the chemical structure. PMID- 10462361 TI - Acute basagran poisoning mimicking neuroleptic malignant syndrome. AB - A 27-year-old robust man, without any medical and surgical history, attempted to commit suicide by consumption of 300 cc (44.1%, 132.3 g) basagran, a readily available herbicide. This poisoning resulted in vomiting, fever, sweating, pipe like muscle rigidity, sinus tachycardia, drowsiness, leukocytosis, rhabdomyolysis and hepatorenal damage. Emperical treatment with bromocriptine was temporally associated with resolution of above signs and symptoms. His clinical presentations and the effect of bromocriptine may be indicative that basagran poisoning mimicks neuroleptic malignant syndrome. PMID- 10462362 TI - Pathogenesis of chronic hepatitis C: immunological features of hepatic injury and viral persistence. AB - The immune response to viral antigens is thought to be responsible for viral clearance and disease pathogenesis during hepatitis C virus (HCV) infection. In chronically infected patients, the T-cell response to the HCV is polyclonal and multispecific, although it is not as strong as the response in acutely infected patients who display a more vigorous T-cell response. Importantly, viral clearance in acutely infected patients is associated with a strong CD4(+) helper T-cell response. Thus, the dominant cause of viral persistence during HCV infection may be the development of a weak antiviral immune response to the viral antigens, with corresponding inability to eradicate infected cells. Alternatively, if clearance of HCV from the liver results from the antiviral effect of T-cell-derived cytokines, as has been demonstrated recently for the hepatitis B virus, chronic HCV infection could occur if HCV is not sensitive to such cytokines or if insufficient quantities of cytokines are produced. Liver cell damage may extend from virally infected to uninfected cells via soluble cytotoxic mediators and recruitment and activation of inflammatory cells forming the necroinflammatory response. Additional factors that could contribute to viral persistence are viral inhibition of antigen processing or presentation, modulation of the response to cytotoxic mediators, immunological tolerance to HCV antigens, mutational inactivation of cytotoxic T lymphocyte (CTL) epitopes, mutational conversion of CTL epitopes into CTL antagonists, and infection of immunologically privileged tissues. Analysis of the basis for viral persistence is hampered because the necessary cell culture system and animal model to study this question do not yet exist. PMID- 10462364 TI - Prognosis of biliary atresia in the era of liver transplantation: French national study from 1986 to 1996. AB - Since the sequential treatment of Kasai operation with or without liver transplantation became available, the overall prognosis of biliary atresia remains unclear. This study examined the prognostic factors from diagnosis. All patients with biliary atresia living in France and born in the years 1986 to 1996 were reviewed. Actuarial survival rates were calculated for survival with native liver, survival after liver transplantation, and overall survival. Potential prognostic factors were analyzed using the logrank test and the Cox model. A total of 472 patients were identified. Ten-year overall survival was 68%. Independent prognostic factors for overall survival were (S = 10-year rates) performance of Kasai operation (performed: S = 69%; not performed: S = 50%), age at Kasai operation (45 days: S = 66%), anatomical pattern of extrahepatic bile ducts, polysplenia syndrome, experience of the center (/=20 [1 center]: S = 78%). Survival with native liver depended on the same independent prognostic factors. In conclusion (1) Kasai operation remains the first line treatment of BA, and (2) early performance of Kasai operation and treatment in an experienced center reduces the need for liver transplantation in infancy and childhood and provides children with the best chance of survival. PMID- 10462363 TI - The effect of ursodeoxycholic acid on the florid duct lesion of primary biliary cirrhosis. AB - The frequency with which florid duct lesions are seen in needle-biopsy specimens of the liver was assessed in patients with primary biliary cirrhosis (PBC) enrolled in a 2-year randomized, double-blind, placebo-controlled trial of ursodeoxycholic acid (UDCA) versus placebo. Paired biopsy specimens obtained at entry and after 2 years on medication were reviewed blindly and mostly simultaneously by a panel of 5 hepatopathologists who, earlier, had characterized the florid duct lesion, which has been well described in the pathology literature. Florid duct lesions at entry were identified in approximately 36%. Patients with earlier disease showed florid duct lesions much more frequently than those with more advanced disease. The prevalence of florid duct lesions in 60 patients receiving placebo medication fell from 38.3% to 21.7%, P =. 025, over the period of 2 years. The prevalence of florid duct lesions also decreased in the 55 patients receiving UDCA, from 32.7% to 18.2%, P =.046. The prevalences of these lesions in the placebo and UDCA patients at entry and at 2 years were not significantly different from each other. The findings suggest that UDCA does not prevent ongoing bile duct destruction in patients with PBC. Instead, they support the impression that UDCA exerts its beneficial effects by protecting against the consequences of bile duct destruction. PMID- 10462365 TI - Transjugular intrahepatic portosystemic shunt compared with endoscopic treatment for prevention of variceal rebleeding: A meta-analysis. AB - Endoscopic treatment (ET) is frequently used to prevent variceal rebleeding but this still occurs in about 50% of patients. Recently, transjugular intrahepatic portosystemic shunt (TIPS) has been compared with ET in several trials. Using a meta-analysis, we evaluated randomized trials comparing TIPS to ET assessing prevention of rebleeding, survival, and the effects on resource use and the quality of patients' lives. Medical databases were searched between January 1988 and January 1999 as well as published citations and conference proceedings. Sensitivity analyses for type of publication, methodological quality score, mean duration of follow-up, type of ET, etiology, and severity of liver disease were performed. Eleven randomized trials involving 811 patients fulfilled the selection criteria. The median follow-up ranged from 10 to 32 months. Variceal rebleeding was significantly more frequent with ET (47%) compared with TIPS (19%) (odds ratio [OR], 3.8; 95% confidence interval [CI], 2.8-5.2; P <.001), but there was no difference in mortality (OR, 0.97; 95% CI, 0.71-1.34). Post-treatment encephalopathy occurred significantly less often after ET (19%) than after TIPS (34%) (OR, 0.43; 95% CI, 0.30-0.60; P <.001). In the studies showing resource use this was more extensive for TIPS. The sensitivity analyses did not alter the main conclusion, and sole comparison with endoscopic ligation did not alter these results. In conclusion, in patients with variceal bleeding, TIPS compared with ET reduces the rebleeding rate, but does not improve survival, and increases the incidence of encephalopathy in a period of 1 to 2.5 years. Thus, TIPS cannot be recommended as the first choice treatment for prevention of variceal rebleeding. PMID- 10462366 TI - Inhibition of 11beta-hydroxysteroid dehydrogenase by bile acids in rats with cirrhosis. AB - Renal sodium retention and potassium loss occur early, in many instances in the preascitic state of cirrhosis, an observation that cannot be fully explained by increased aldosterone concentrations. We therefore hypothesize that 11beta hydroxysteroid dehydrogenase 2 (11beta-HSD2), which protects mineralocorticoid receptors (MR) from glucocorticosteroids, is down-regulated in cirrhosis. Cirrhosis was induced by bile duct ligation in rats. The urinary ratio of (tetrahydrocorticosterone + 5alpha-tetrahydrocorticosterone)/ 11-dehydro tetrahydrocorticosterone [(THB+5alpha-THB)/THA] was measured by gas chromatography. Cortical collecting tubules (CCT) were isolated by microdissection and used for measurements of the activity of 11beta-HSD2 by assessing the conversion of corticosterone to dehydrocorticosterone. The mRNA content of 11beta-HSD2 was determined by reverse-transcription polymerase chain reaction (RT-PCR) in CCTs. The urinary ratio of (THB+5alpha-THB)/THA increased concomitantly with the urinary excretion of bile acids following bile duct ligation. Chenodeoxycholic acid (CDCA) dose-dependently inhibited 11beta-HSD2 in CCT with a Ki of 19.9 micromol/L. Four weeks after bile duct ligation, 11beta HSD2 activity was decreased in CCT, an observation preceded by a reduced mRNA content at weeks 2 and 3. In cirrhosis, the MR-protecting effect by 11beta-HSD2 is diminished, and therefore, endogenous glucocorticoids can induce MR-mediated sodium retention and potassium loss. PMID- 10462367 TI - Effects of vasopressin on portal-systemic collaterals in portal hypertensive rats: role of nitric oxide and prostaglandin. AB - This study investigated the effect of vasopressin on portal-systemic collaterals in portal hypertensive rats and the influence of nitric oxide (NO) and prostaglandin on the responsiveness of collateral vessels to vasopressin. The vascular responsiveness to graded concentrations of vasopressin was tested with or without the incubation of n(omega)-nitro-L-arginine (NNA) (100 micromol/L) and/or indomethacin (10 micromol/L) in perfused collateral vascular beds of rats with portal hypertension induced by partial portal vein ligation. In addition, concentration-response curves to vasopressin with incubation of a vasopressin V(1) receptor antagonist d(CH(2))(5)Tyr(Me) arginine vasopressin and concentration-response curves to a V(2) receptor agonist 1-desamino-8-D-arginine vasopressin were performed. Vasopressin significantly increased the perfusion pressure of collaterals, and this effect was suppressed by the addition of the V(1) receptor antagonist. Perfusion with the V(2) receptor agonist had no effect on the collaterals. Incubation with NNA, indomethacin, or both significantly potentiated the response of collaterals to vasopressin. In addition, the pressor response to vasopressin in the combination group was significantly higher than that in the NNA-alone group. The results show that vasopressin produces a direct vasoconstrictive effect on the portal-systemic collaterals of portal hypertensive rats. This effect is mediated by the vasopressin V(1,) but not V(2), receptors. The attenuation of the response to vasopressin by NO and prostaglandin suggest a function role of both mediators in the regulation of the portal-systemic collateral circulation in portal hypertensive rats. PMID- 10462368 TI - L-ornithine-L-aspartate lowers plasma and cerebrospinal fluid ammonia and prevents brain edema in rats with acute liver failure. AB - Brain edema sufficient to cause intracranial hypertension and brain herniation remains a major cause of mortality in acute liver failure (ALF). Studies in experimental animal models of ALF suggest a role for ammonia in the pathogenesis of both encephalopathy and brain edema in this condition. As part of a series of studies to evaluate the therapeutic efficacy of ammonia-lowering agents, groups of rats with ALF caused by hepatic devascularization were treated with L ornithine-L-aspartate (OA), an agent shown previously to be effective in reducing blood ammonia concentrations in both experimental and human chronic liver failure. Treatment of rats in ALF with infusions of OA (0.33 g/kg/h, intravenously) resulted in normalization of plasma ammonia concentrations and in a significant delay in onset of severe encephalopathy. More importantly, brain water content was significantly reduced in OA-treated rats with ALF. These protective effects of OA were accompanied by increased plasma concentrations of several amino acids including glutamate, gamma-aminobutyric acid (GABA), taurine, and alanine, as well as the branched-chain amino acids, leucine, isoleucine, and valine. Increased availability of glutamate following OA treatment provides the substrate for the major ammonia-removal mechanism (glutamine synthetase). Plasma (but not cerebrospinal fluid) glutamine concentrations were increased 2-fold (P <.02) in OA-treated rats, consistent with increased muscle glutamine synthesis. Direct measurement of glutamine synthetase activities revealed a 2-fold increase following OA treatment. These findings demonstrate a significant ammonia-lowering effect of OA together with a protective effect on the development of encephalopathy and brain edema in this model of ALF. PMID- 10462369 TI - Improvement in pulmonary hemodynamics during intravenous epoprostenol (prostacyclin): A study of 15 patients with moderate to severe portopulmonary hypertension. AB - Pulmonary hypertension associated with increased pulmonary vascular resistance (PVR) and occurring in the setting of portal hypertension is referred to as "portopulmonary hypertension." Intravenous epoprostenol (prostacyclin) is a potent pulmonary and systemic vasodilator with antithrombotic properties. It can decrease PVR and pulmonary artery pressure in patients with primary (idiopathic) pulmonary hypertension. Using right-heart catheterization, we evaluated the acute pulmonary hemodynamic effects of intravenous epoprostenol in patients with moderate to severe pulmonary hypertension (mean pulmonary artery pressure [MPAP] >/=35 mm Hg) associated with clinical manifestations of portal hypertension. Effects of long-term infusion of epoprostenol were also evaluated. We studied 15 consecutive patients with portopulmonary hypertension; 14 underwent acute administration of epoprostenol, and no significant side effects were noted. Ten patients received continuous epoprostenol (range, 8 days-30 months). Acute changes in PVR (-34% +/- 18%), MPAP (-16% +/- 10%), and cardiac output (CO) (+21 +/- 18%), were statistically significant (P <.01). Long-term use of epoprostenol further lowered PVR (-47% +/- 12% from baseline and -31% +/- 22% from the acute change; P <.05) in the 6 patients restudied by right-heart catheterization. Death occurred in 6 of 10 (60%) of those receiving long-term epoprostenol. In moderate to severe portopulmonary hypertension, intravenous epoprostenol resulted in a significant improvement (both acute and long-term) in PVR, MPAP, and CO. Potential adverse effects on portal hypertension and implications for orthotopic liver transplantation (OLT), however, require further study. PMID- 10462370 TI - Glucose intolerance and insulin resistance in cirrhosis are normalized after liver transplantation. AB - Cirrhosis is often associated with insulin resistance and glucose intolerance. We evaluated if these alterations are restored by liver transplantation (LT). Glucose tolerance (oral glucose tolerance test [OGTT]), peripheral insulin sensitivity (euglycemic insulin clamp technique), glucose oxidation (indirect calorimetry), nonoxidative glucose disposal, and insulin secretion (hyperglycemic clamp technique) were measured in 6 patients (Group 1) before and 6 months after LT, in 12 patients (Group 2) who underwent LT 6 to 30 months previously, and in 6 healthy individuals (controls). In Group 1, glucose tolerance and insulin sensitivity (3.24 +/- 0.37 mg/kg/min) were normalized after LT (8.6 +/- 0.77 mg/kg/min; P <.0001; P = not significant vs. controls). The improved insulin mediated glucose uptake was the result of a normalization of nonoxidative glucose disposal. Fasting insulin and C-peptide decreased from 24.6 +/- 3.3 microU/mL and 4.37 +/- 0.46 ng/dL, respectively, to 12.7 +/- 1.9 microU/mL and 2.46 +/- 0.5 ng/dL (controls: 10.0 +/- 3 microU/mL and 1.45 +/- 0.34 ng/dL). The glucose induced increase of insulin concentration, which was higher before LT, showed a significant reduction, although the first phase of beta-cell secretion remained significantly higher compared with that of controls. All these findings were also confirmed in Group 2. The present data indicate that LT normalizes glucose tolerance and insulin sensitivity in cirrhotic patients through an improvement of both hepatic glucose clearance and the peripheral glucose disposal. The latter effect may be the result of the correction of chronic hyperinsulinemia. An increased first-phase beta-cell insulin secretion in response to high glucose levels persists, suggesting that a memory of previous insulin resistance is maintained. PMID- 10462371 TI - Resting energy expenditure should be measured in patients with cirrhosis, not predicted. AB - Measurements of resting energy expenditure (REE) can be used to determine energy requirements. Prediction formulae can be used to estimate REE but have not been validated in cirrhotic patients. REE was measured, by indirect calorimetry, in 100 cirrhotic patients and 41 comparable healthy volunteers, and the results compared with estimates predicted using the Harris-Benedict, Schofield, Mifflin, Cunningham, and Owen formulae, and the disease-specific Muller formula. The mean (+/- 1 SD) measured REE in the healthy volunteers (1,590 +/- 306 kcal/24 h) was significantly greater than the mean Harris-Benedict, Mifflin, Cunningham, and Owen predictions but comparable with the mean Schofield prediction; individual predicted values varied widely from measured values (95% limits of agreement, 460 to +424 kcal). The mean measured REE in the cirrhotic patients was significantly greater than in the healthy volunteers (23.2 +/- 3. 8 cf 21.9 +/- 2.9 kcal/kg/24 h; P <.05). The mean measured REE in the cirrhotic patients (1,660 +/- 337 kcal/24 h) was significantly different from mean predicted values (Harris Benedict, 1,532 +/- 252 kcal/24 h, P <.0001; Schofield, 1,575 +/- 254 kcal/24 h, P <.0005; Mifflin, 1,460 +/- 254 kcal/24 h, P <.0001; Cunningham, 1,713 +/- 252 kcal/24 h, P <.05; Owen, 1,521 +/- 281 kcal/24 h, P <.0001; Muller, 1,783 +/- 204 kcal/24 h, P <.0001); individual predicted values varied widely from measured values (95% limits of agreement, -632 to +573 kcal). Simple regression analysis showed that fat-free mass (FFM) was the strongest predictor of measured REE in the cirrhotic patients, accounting for 52% of the variation observed. However, a population-specific prediction equation, derived using stepwise regression analysis, which incorporated FFM, age, and Pugh's score, accounted for only 61% of the observed variation in measured REE. REE should, therefore, be measured in cirrhotic patients, not predicted. PMID- 10462372 TI - Prevalence of the C282Y mutation of the hemochromatosis gene in liver transplant recipients and donors. AB - Hemochromatosis heterozygotes may be predisposed to end-stage liver disease from other causes. The aims of this study were to determine the prevalence of the hemochromatosis mutation, C282Y, in end-stage liver disease and to determine if transplantation of C282Y heterozygous livers adversely affected survival. The C282Y status of patients who underwent hepatic transplantation and, whenever possible, their donors, was determined and correlated with hepatic iron status. Survival was compared in patients who received livers from heterozygotes and normals. Prevalence of C282Y in recipients was compared with 5,211 voluntary blood donors. Twenty-six C282Y heterozygotes were detected among 304 transplant recipients (8.6%) compared with a prevalence of 8.4% in blood donors. Six of 26 heterozygous recipients (23%) had >/=2+ iron staining in their explanted livers compared with 40 of 277 wild-type livers (14%) (P = ns). There was no significant difference in mean hepatic iron concentration between C282Y heterozygotes and wild-type explanted livers with >/=2+ iron staining. Seven of 31 patients (23%) with alcoholic liver disease were C282Y heterozygotes. Twenty-four heterozygotes were detected in 141 donors (17.0%). Survival did not differ between recipients who received heterozygous or normal livers. The prevalence of C282Y heterozygotes in patients requiring liver transplantation does not differ significantly from the general population. Heterozygotes are not at increased risk of developing end stage liver disease. Transplantation of C282Y heterozygous livers is a safe, effective practice. PMID- 10462373 TI - Nitric oxide production and inducible nitric oxide synthase expression in peritoneal macrophages of cirrhotic patients. AB - The present study assessed whether peritoneal macrophages isolated from cirrhotic patients produce nitric oxide (NO) and express NO synthase type II (NOS II) mRNA and protein. Patients with cirrhosis and ascites without peritonitis or with unresolved or resolved spontaneous bacterial peritonitis (SBP) were studied. Following paracentesis, ascites NO(2)(-) + NO(3)(-) content (NOx) was measured. Peritoneal macrophages from ascites were seeded on well plates, and NO(2)(-) in the medium was determined. NOx was higher in patients with unresolved or resolved SBP than in cirrhotic patients without peritonitis. Macrophages of patients with SBP or resolved SBP produced NO(2)(-) after 30 hours in culture, but those obtained from patients without peritonitis did not. Reverse-transcription polymerase chain reaction (RT-PCR) and immunocytochemical analysis revealed the presence of a clear signal for NOS II mRNA and protein in macrophages of SBP patients, regardless of whether or not the infection subsided. Therefore, peritoneal macrophages isolated from cirrhotic patients with unresolved or resolved SBP produce NO and express the NOS II mRNA and protein, suggesting that NOS II may contribute to the control of SBP, or to its associated pathology, in human cirrhosis. PMID- 10462374 TI - Structure and expression of Tg737, a putative tumor suppressor gene, in human hepatocellular carcinomas. AB - Deletions of the Tg737 gene, whose product is involved in liver oval cell proliferation, differentiation, and ploidy control, have been recently shown in chemically induced rat liver tumors and in a limited series of patients with hepatocellular carcinoma (HCC). Thus, Tg737 has been proposed as a candidate new liver-specific tumor suppressor gene. To investigate this important issue, we analyzed the structure and expression pattern of the Tg737 gene in a group of 23 tumorous and adjacent nontumorous liver tissues, by combining polymerase chain reaction (PCR) and Southern and Northern blot-based analyses. We failed to identify deletions or gross alterations of the Tg737 gene by both PCR and Southern blot analyses. Northern blots showed comparable accumulation of normal Tg737 transcripts in both tumorous and nontumorous tissues. Collectively, therefore, our results do not support the hypothesis of frequent Tg737 genetic alterations in human HCC. PMID- 10462375 TI - Modulation of hepatic acute phase gene expression by epidermal growth factor and Src protein tyrosine kinases in murine and human hepatic cells. AB - As part of systemic inflammatory reactions, interleukin 6 (IL-6) induces acute phase protein (APP) genes through the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway. Epidermal growth factor (EGF), which contributes to the regenerative process after liver injury and also activates STATs, does not induce but attenuates IL-6-stimulated expression of several APP genes in primary mouse hepatocytes. The APP-modifying action of EGF receptor (EGFR) was characterized in HepG2 cells. Although EGF less effectively engages STAT proteins in these cells, it reduces expression of fibrinogen and haptoglobin, but stimulates production of alpha(1)-antichymotrypsin and induces transcription through the alpha(1)-antichymotrypsin and C-reactive protein promoter. The stimulatory EGFR signal is insensitive to inhibition of JAKs and appears to involve Src kinases and STAT proteins as shown by inhibition through overexpression of C-terminal Src kinase (Csk) and transdominant negative STAT3, respectively. A mediator role of Src is supported by the ability of c-Src and v Src to activate STATs and induce transcription through APP promoters. Src kinases have been observed in association with the IL-6 receptor; however, inhibition of Src kinases by Csk enhances IL-6-induced transcription. The Csk effect is attributed to prevention of Src kinases from phosphorylating gp130 at the docking site for the signal-moderating protein tyrosine phosphatase SHP-2. The inhibitory EGFR signal on APP expression correlates with the activation of Erk1 and Erk2. The study shows a dual signaling function for EGFR and suggests that the ratio of receptor-activated STATs and Erks influence the level of stimulated or inhibited expression of individual APPs. PMID- 10462376 TI - Abnormal regulation of aortic NOS2 and NOS3 activity and expression from portal vein-stenosed rats after lipopolysaccharide administration. AB - Hyporeactivity to vasoconstrictors in aortae from portal vein-stenosed rats is associated with an increased activity of endothelial NO synthase (NOS3). In contrast, during sepsis, which is common in cirrhosis, vascular hyporeactivity is associated with an induction of inducible NOS2. The aim of this study was to investigate the in vitro reactivity to phenylephrine and the regulation of NOS2 and NOS3 in aortae from portal vein-stenosed rats after lipopolysaccharide (LPS) administration. Aortic vascular reactivity for phenylephrine, aortic NOS activity, and NOS2 and NOS3 protein expression were determined 5 hours after intravenous LPS or saline administration. Moreover, aortic NOS activity was measured after 5-hour in vitro incubation in LPS. LPS induced a significantly smaller decrease in aortic tension in portal vein-stenosed than in sham-operated rats. Under baseline conditions, aortic NOS activity and NOS3 protein expression were higher in portal vein-stenosed than in sham-operated rats, and NOS2 protein expression was not detected in aortae from either group. After LPS administration, NOS activity and NOS2 protein expression increased significantly less in portal vein-stenosed than in sham-operated rat aortae. Similar results were obtained after in vitro incubation with LPS. Endothelium removal or NOS3 inhibition with the calmodulin inhibitor, W7, increased NOS activity in the aortae of portal vein-stenosed rats after LPS incubation. In conclusion, in aortae of portal vein-stenosed rats exposed to LPS, no further decrease in aortic reactivity to phenylephrine was observed, and the induction of NOS2 was down regulated. Endothelium removal or calmodulin inhibition inhibits NOS3 overactivity and leads to normalized NOS2 activation after LPS in aortae from portal vein-stenosed rats. PMID- 10462377 TI - Consistent infiltration of thymus-derived T cells into the parenchymal space of the liver in normal mice. AB - We previously reported that extrathymic T cells (intermediate T-cell receptor cells [TCR(int) cells]) are in situ generated in the parenchymal space of the liver in mice. They subsequently migrate to the sinusoidal lumen. In this study, we characterized how such extrathymic T cells, natural killer (NK) cells, and thymus-derived T cells (high T-cell receptor cells [TCR(high) cells]) localized in the parenchymal space or the sinusoidal lumen of mice. To this end, liver irrigation with physiological saline from the portal vein was performed and the distribution of lymphocyte subsets was compared between the liver (i.e., lymphocytes in the parenchymal space) and the irrigation solution (i.e., lymphocytes in the sinusoidal lumen). Extrathymic T cells and NK cells were found to be abundant in both the liver and sinusoidal lumen. As expected, thymus derived T cells were abundant in the sinusoidal lumen. However, a significant proportion of thymus-derived T cells were always present in the parenchymal space, even after intensive irrigation with or without collagenase. These results suggest that thymus-derived T cells may consistently infiltrate the parenchymal space from the sinusoidal lumen in normal mice. This possibility was confirmed by (1) the injection of B6 splenic cells (TCR(high) cells) or the thymus graft into B6-nu/nu mice (presence of only TCR(int) cells) and by (2) using parabiotic mice of B6.Ly5.1 and B6.Ly5.2 strains (sharing circulation) in conjunction with immunofluorescence tests and immunohistochemical staining. In other words, inverted routes of migration and homing between extrathymic T cells and thymus derived T cells exist in the liver. PMID- 10462378 TI - Redox manipulation using the thiol-oxidizing agent diethyl maleate prevents hepatocellular necrosis and apoptosis in a rodent endotoxemia model. AB - Manipulation of the intracellular redox state has been shown to alter cell activation pathways with resultant changes in cellular function. Previous studies have suggested that thiol oxidation, using the glutathione-depleting agent diethyl maleate (DEM), was able to inhibit endothelial cell activation. We hypothesized that this agent might exert beneficial effects following endotoxemia in the rat, a model in which transendothelial migration of neutrophils is central to the development of hepatocellular injury. Sprague-Dawley rats treated intraperitoneally with lipopolysaccharide (LPS) (200 microg/kg) plus D galactosamine (GalN) (600 mg/kg) developed hepatocellular necrosis, as evidenced by liver enzyme release and morphological changes. Pretreatment with DEM abrogated this injury in a dose-dependent fashion. Histology revealed reduced neutrophil accumulation in both the parenchyma and sinusoids, consistent with reduced neutrophil sequestration and transendothelial migration. This effect appeared to be related to the ability of DEM to prevent LPS-induced up-regulation of both vascular cell adhesion molecule-1 (VCAM-1) mRNA and intercellular adhesion molecule-1 (ICAM-1) mRNA in the liver, as well as reducing tumor necrosis factor (TNF) mRNA expression. In addition, DEM prevented hepatocyte apoptosis following LPS treatment. The effect was reproduced when TNF was used as an inflammatory stimulus, suggesting a direct protective effect on the hepatocyte. Taken together, these studies show that redox manipulation through thiol oxidation may represent a novel approach to preventing liver necrosis and apoptosis in inflammatory conditions. PMID- 10462379 TI - The unique acyl chain specificity of biliary phosphatidylcholines in mice is independent of their biosynthetic origin in the liver. AB - The liver synthesizes phosphatidylcholine (PC) de novo from choline via the CDP choline pathway, and from phosphatidylethanolamine (PE) via the phosphatidylethanolamine N-methyltransferase (PEMT) pathway. Significant amounts of PC, which are highly specific in their acyl chain composition, are secreted into bile by the liver. To determine whether either of the 2 PC biosynthetic routes is sufficient to provide physiological PC concentrations in bile, or is responsible for the unique acyl chain composition of bile PC, we analyzed gallbladder bile composition in mice that synthesized PC either via the PEMT pathway (induced by feeding a choline-deficient diet) or the CDP-choline pathway (based on genetic PEMT-deficiency). The PC concentration in gallbladder bile of mice that synthesize PC mainly via the CDP-choline pathway was comparable with control mice that synthesize PC via both pathways, whereas it was reduced by approximately 40% in mice that synthesize PC via the PEMT pathway. The acyl chain composition of bile PC was similar irrespective of the active PC biosynthetic pathway in the liver. These data demonstrate that the CDP-choline pathway alone, but not the PEMT pathway alone, can account for physiological concentrations of PC in gallbladder bile. Moreover, the specificity of biliary PC fatty acyl composition is determined independently from the synthetic origin of PC. PMID- 10462380 TI - Bile acid structure and selective modulation of murine hepatic cytochrome P450 linked enzymes. AB - We examined the effects of the administration of different bile acids on in vivo hepatic murine cytochrome P450 (CYP) content, nicotinamide adenine dinucleotide phosphate (NADPH)-CYP-reductase, and individual mixed-function oxidases (MFOs). Neither CYP level nor reductase were appreciably affected by single intraperitoneal administration of taurodeoxycholic acid (TDCA) (12.2 or 24.4 mg x kg(-1) bw). MFO to various isoenzymes were slightly reduced 24 hours after treatment. Taurohyodeoxycholic acid (THDCA) and tauroursodeoxycholic acid (TUDCA) both induced CYP, reductase, and MFOs. CYP3A1/2-linked activity (i.e., testosterone 6beta-hydroxylase, and N-demethylation of aminopyrine) in a dose dependent fashion was enhanced ( approximately 2-3-fold). CYP2E1- (hydroxylation of p-nitrophenol), CYP1A2-(O-demethylation of methoxyresorufin), CYP2A1/2- and CYP2B1/2-(6alpha-hydroxylase), and CYP2B9- (16alpha-hydroxylase) dependent MFOs, as well as 7alpha-, 16beta-, 2alpha-, and 2beta-hydroxylations, were all significantly induced by THDCA. Apart from alkoxyresorufin metabolism and a modest CYP2E1 increase, TUDCA behaved like THDCA. A generalized induction was also recorded after ursodeoxycholic acid (UDCA) administration. THDCA and TDCA did not show substantial differences in the N-demethylation of aminopyrine when different species (rat vs. mouse) and administration route (intraperitoneal vs. intravenous) were compared. Results on the most affected isoenzymes, CYP3A1/2 (THDCA, TUDCA, and UDCA) and CYP2E1 (UDCA), were sustained by means of Western immunoblotting. CYP3A induction was paralleled by a corresponding increase in mRNA. These data could partially explain the therapeutic mechanism of UDCA, TUDCA, and THDCA in chronic cholestatic liver disease. CYP3A induction, which is linked to P-glycoprotein (Pgp) family overexpression, may enhance hepatic metabolism, transport, and excretion of toxic endogenous lipophilic bile acids. PMID- 10462381 TI - Deficient assembly and function of gap junctions in Trf1, a trafficking mutant of the human liver-derived cell line HuH-7. AB - The Trf1 cell line, selected from the human hepatoma cell line HuH-7, manifests altered trafficking of various plasma membrane proteins. In particular, there is a striking loss of State 2 asialoglycoprotein receptors. This cell line is shown here to also manifest defects in function and assembly of gap junctions comprising connexin43 (Cx43). No alteration of Cx43 expression or phosphorylation was apparent. Nevertheless, immunostaining of Cx43 revealed that fewer and smaller gap junctions were present at appositional membrane areas in Trf1 cells as compared with parental HuH-7. This correlated with a significant attenuation in gap junction-mediated communication between Trf1 cells as demonstrated by markedly decreased dye transfer and their reduced ability to propagate mechanically evoked Ca(2+) waves. Isoelectric focusing (IEF) of Cx43 in HuH-7 cells indicated that the pIs of this protein were significantly lower than that predicted from its amino acid sequence; no differences in pI were evident in Cx43 from Trf1 cells and the HuH-7 cell line. The effects of the Trf1 mutation on assembly and function of gap junctions indicate that this mutation influences trafficking of Cx43. Connexins differ in several respects from other membrane proteins thus far analyzed in Trf1 mutants: gap junctions localize exclusively to the lateral cell surface; they are not glycoproteins; and they do not play a role in endocytic pathways. The disruption of trafficking of Cx43 by this mutation suggests that the Trf1 phenotype is a defect at a common point along the trafficking pathway of cell-surface proteins, irrespective of their ultimate destination on the cell surface or their glycosylation profile. PMID- 10462382 TI - Targeting of aminopeptidase N to bile canaliculi correlates with secretory activities of the developing canalicular domain. AB - We have used human hepatoma cell lines as an in vitro model to study the development of hepatic bile canaliculi (BC). Well-differentiated hepatoma cells cultured for 72 hours could develop characteristic spheroid structures at sites of cell-cell contact that contained tight junctions and various membrane protein markers, resembling BC found in vivo. Intact cytoskeleton was essential for this differentiation process. In the coculture experiments in which cells of different origins were populated together, BC only formed between hepatic cells and preferentially among well-differentiated cells. Poorly differentiated hepatoma cells never formed BC among themselves, but could be induced to undergo canalicular differentiation by interacting with well-differentiated cells. During BC morphogenesis, integral canalicular membrane proteins were gradually delivered and accumulated at the developing BC. Among them, targeting of aminopeptidase N (APN) seemed to correlate with activation of certain secretory functions. Specifically, only APN-positive BC supported excretion of fluorescein diacetate (FDA) and 70-kd dextran, but had no relationship with secretion of horseradish peroxidase (HRP). Targeting of another BC protein, dipeptidyl peptidase IV (DPPIV), on the other hand, bore no association with any secretory activity examined. In addition, inhibition of enzymatic activity of APN could perturb canalicular differentiation without affecting cell proliferation. Our results suggest that targeting of APN proteins may reflect or even play an important role in the development and functional maturation of the canalicular structures. PMID- 10462383 TI - Persistent activation of nuclear factor-kappaB in cultured rat hepatic stellate cells involves the induction of potentially novel Rel-like factors and prolonged changes in the expression of IkappaB family proteins. AB - Rat hepatic stellate cells (HSC) cultured in serum-containing medium underwent a rapid (3-hour) classical induction of p50:p65 and p65:p65 nuclear factor-kappaB (NF-kappaB) dimers. Subsequent culturing was associated with prolonged expression of active p50:p65 and persistent induction of a high-mobility NF-kappaB DNA binding complex consisting of potentially novel Rel-like protein(s). Formation of the latter complex was competed for by specific double-stranded oligonucleotides, was up-regulated by treatment of HSCs with tumor necrosis factor alpha (TNF alpha), and was maintained at basal levels of expression by a soluble HSC-derived factor. An NF-kappaB-responsive CAT reporter gene was highly active in early cultured HSCs but was also trans-activated at a lower but significant level in longer-term cultured cells and could be completely suppressed by expression of dominant negative IkappaB-alpha. Physiological significance of the lower persistent NF-kappaB activities was also demonstrated by the ability of long-term cultured HSCs to support the activity of the NF-kappaB-dependent human intercellular adhesion molecule-1 (ICAM-1) promoter. Freshly isolated HSCs expressed high levels of IkappaB-alpha and IkappaB-beta. Culture activation was accompanied by a long-term reduction in levels of IkappaB-alpha with no detectable expression in the nuclear fraction of cells, under these conditions p50:p65 was detected in the nucleus. IkappaB-beta expression was transiently reduced and, upon replenishment, was associated with appearance of a lower mobility IkappaB-beta antibody-reactive species. Bcl3 expression was absent in freshly isolated HSC but was induced during culturing and became a persistent feature of the activated HSC. Inhibition of NF-kappaB DNA binding activity by gliotoxin was associated with increased numbers of apoptotic cells. We suggest that activation of NF-kappaB in cultured HSC is required for expression of specific genes associated with the activated phenotype such as ICAM-1 and may be antiapoptotic for rat HSCs. PMID- 10462384 TI - Pretherapy alanine transaminase level as a determinant for hepatitis B e antigen seroconversion during lamivudine therapy in patients with chronic hepatitis B. Asian Hepatitis Lamivudine Trial Group. AB - In the reported Asian lamivudine trial, the rate of hepatitis B e antigen (HBeAg) seroconversion, defined as HBeAg/hepatitis B virus (HBV) DNA seroclearance and development of anti-HBe, during 52 weeks of treatment was only 13% to 16%. To evaluate whether any factors influenced HBeAg seroconversion, data from 345 patients in that trial were reanalyzed to correlate HBeAg seroconversion with variables including treatment, age, gender, body build, histology, baseline HBV DNA levels, and alanine transaminase (ALT) levels. Exploratory analysis using stepwise modeling revealed that HBeAg seroconversion correlated highly with pretherapy ALT (P <.001) followed by lamivudine therapy (P =.013), but only marginally with baseline HBV-DNA (P =.071) and cirrhosis (P =.066) for lamivudine 100 mg and placebo comparison. Among these four variables, only pretherapy ALT still had a highly significant (P <.001) correlation and lamivudine therapy had a borderline association (P =.066) for lamivudine 25 mg and placebo comparison. Categorical analysis revealed that HBeAg seroconversion occurred earlier and the cumulative rate was significantly higher in patients with pretherapy ALT values over 2 times the upper limit of normal (ULN) as compared with treated patients with lower ALT levels or untreated control patients with the same ALT levels (P <.001, respectively). The highest HBeAg seroconversion rate was observed in 100 mg lamivudine-treated patients with ALT levels greater than 5 times the ULN (64%) compared with patients with ALT 2 to 5 times the ULN (26%, P =.03); and ALT less than 2 times the ULN, (5%, P <.001). These results suggest that pretherapy ALT is the strongest determinant for HBeAg seroconversion during lamivudine therapy, and should be considered in selecting patients for treatment. PMID- 10462385 TI - Role of hepatitis C virus in lymphoproliferative disorders after liver transplantation. AB - It has been suggested that hepatitis C virus (HCV) infection could be associated with B-cell clonal expansion. The aim of this study was to analyze the relationship between lymphoproliferative disorders and HCV infection in liver transplant recipients. We studied 157 patients receiving a liver transplant between January 1989 and May 1997 with a follow-up longer than 3 months. The incidence of posttransplant lymphoproliferative disorders (PTLDs) was analyzed with reference to the indication for liver transplantation, the induction and maintenance immunosuppression, the incidence of acute rejection episodes, and Epstein-Barr virus (EBV) infection. Six PTLDs occurred after a median posttransplant follow-up of 7 months (3.8%). Four of the 6 PTLDs occurred among the 38 patients transplanted for HCV-related cirrhosis, and 2 PTLDs occurred in the 119 patients receiving a liver transplant for non-HCV liver diseases (10.5% vs. 1.7%, respectively; P =.03). The 4-year probability of PTLD was significantly higher in patients receiving a liver transplant for HCV-related cirrhosis than non-HCV liver diseases (12.3% vs. 2.2%, respectively; P =.015). Patients receiving a liver transplant for HCV-related cirrhosis were more likely to receive antithymocyte globulins (ATG). However, in patients treated with ATG, the 4-year probability of PTLD was higher among those patients receiving a liver transplant for HCV-related cirrhosis than for non-HCV liver diseases (27.1% vs. 6.4%, respectively; P =.08). EBV gene products were detected in tumor tissues in 3 of 4 patients with HCV-associated PTLD. Our data suggest that, in addition to EBV infection, 2 mutually nonexclusive factors, i.e., the use of ATG and HCV infection, could play a role in the occurrence of PTLD after a liver transplant for HCV-related cirrhosis. PMID- 10462386 TI - Licensed recombinant hepatitis B vaccines protect chimpanzees against infection with the prototype surface gene mutant of hepatitis B virus. AB - The emergence in vaccinated individuals of hepatitis B virus (HBV) mutants with amino acid substitutions within the a determinant of the surface protein has raised the possibility that such variants represent neutralization escape mutants. We previously demonstrated that one such mutant HBV, strain AS, with an arginine substituted for glycine at surface gene codon 145, was infectious and pathogenic in seronegative chimpanzees. In the present study, the protective efficacy of licensed hepatitis B vaccines was evaluated against challenge with this mutant virus. Four chimpanzees were immunized with 1 of 2 licensed recombinant hepatitis B vaccines. Shortly after the chimpanzees developed antibodies to hepatitis B surface antigen (anti-HBs), they were challenged intravenously with mutant HBV strain AS. Two unvaccinated chimpanzees served as positive controls. The 4 vaccinated chimpanzees did not develop evidence of HBV infection or hepatitis during 2 years following virus challenge. In contrast, the 2 unvaccinated chimpanzees developed HBV infection and hepatitis. Serum anti-HBs in the vaccinated chimpanzees reacted not only with wild-type surface antigen, but also with mutant surface antigen by competition enzyme-linked immunosorbent assay (ELISA). Thus, immunization of chimpanzees with licensed recombinant hepatitis B vaccines stimulates anti-HBs that is broadly reactive and affords protection against infection with a surface gene mutant of HBV, suggesting that properly immunized individuals are not at significant risk of infection with this prototype variant strain of HBV. PMID- 10462387 TI - Racial differences in responses to therapy with interferon in chronic hepatitis C. Consensus Interferon Study Group. AB - The likelihood of a sustained response to a course of interferon in patients with chronic hepatitis C correlates with several clinical and viral factors, including age, viral genotype and initial levels of hepatitis C virus (HCV) RNA in serum. The role of race and ethnicity has not been assessed. We evaluated the association of race with response to interferon in a large randomized, controlled trial using either consensus interferon (9 microg) or interferon alfa-2b (3 million units) given three times weekly for 24 weeks. African-American patients participating in the study were similar to white patients in mean age (43 vs. 42 years) and baseline levels of HCV RNA (3.6 vs. 3.0 million copies/mL) but had lower rates of cirrhosis (5% vs. 12%) and more frequently had viral genotype 1 (88% vs. 66%: P =.004). Most strikingly, the rates of end-of-treatment and sustained virological responses were lower among the 40 African-American patients (5% and 2%) than among the 380 white patients (33% and 12%) (P =.04 and.07). Rates of response among Hispanic and Asian-American patients were not statistically different than non-Hispanic white patients. Median viral levels decreased by week 24 of therapy by 2.5 logs in white patients (from 3.0 to 0.012 million copies/mL) but by only 0.5 logs among African- American patients (from 3.6 to 1.8 million copies/mL). Thus, there are marked racial differences in virological responses to interferon in hepatitis C that must be considered in assessing trials of interferon therapy and in counseling patients regarding treatment. The differences in response rates are as yet unexplained. PMID- 10462388 TI - Current practice patterns of primary care physicians in the management of patients with hepatitis C. AB - Approximately 4 million Americans are infected with the hepatitis C virus (HCV). Most patients with hepatitis C have no symptoms until cirrhosis is established. Thus, initial diagnosis and management of hepatitis C rely on primary care physicians identifying and screening high-risk individuals. We administered a survey to 1,233 primary care physicians in a health maintenance organization (HMO) in April 1997 to assess their knowledge of the risk factors for HCV infection and approach to the management of 2 hypothetical HCV antibody-positive patients, 1 with elevated and the other with normal alanine transaminase (ALT). Four hundred four (33%) physicians returned the survey. Ninety percent of respondents correctly identified the risk factors for HCV infection, but 20% still considered blood transfusion in 1994 as a significant risk factor for HCV infection. Sixty-two percent of respondents would refer HCV antibody-positive patients with abnormal transaminase levels, but 33% would follow these patients themselves, even though none of the respondents had treated any hepatitis C patient on their own. Forty-three percent of respondents overestimated, while 29% did not know the efficacy of interferon treatment. Sixty-five percent of respondents would retest patients for HCV antibody, regardless of risk factors and transaminase levels. We found that most primary care physicians correctly identified the significant risk factors for HCV infection and appropriately managed the 2 hypothetical patients, but there was considerable confusion about the use of HCV tests and the effectiveness of treatment. Educational programs for primary care physicians are needed to implement hepatitis C screening and to initiate further evaluation and management of those who test positive. PMID- 10462389 TI - Chronic hepatitis C: interferon retreatment of relapsers. A meta-analysis of individual patient data. European Concerted Action on Viral Hepatitis (EUROHEP). AB - Relapse after interferon (IFN) therapy for chronic hepatitis C virus (HCV) infection occurs in 50% of patients after the initial response. The benefit of retreatment with IFN alone has not been assessed in large controlled studies. To assess the effectiveness and the tolerability of IFN retreatment and to identify the optimal second course regimen, we performed a meta-analysis of individual patient's data on a set of 549 patients (mean age 43.8 years; 12.2 SD, men: 65%) who had an end-of-treatment biochemical response to a first IFN course and then relapsed. Retreatment was started within 24 months after the end of the first course. Biochemical end-of-treatment responses (ETR) and sustained responses (SR) were observed in 405 of 549 (73.8%; 95% confidence interval [CI] 70.1-77.5) and in 124 of 549 (22.6%; CI 19.1-26.1) patients, respectively. One hundred seventy five of 404 patients (43.3%; CI 38.6-48.2) developed an end-of-treatment, biochemical, and virological response when retreated. A biochemical and virological SR to retreatment occurred in 73 of 494 (14.8%; CI 11.7-18) patients. Thirty-two patients (5. 8%; CI 3.5-7.8) stopped retreatment for adverse effects. Biochemical and virological SR was predicted independently by logistic regression analysis using a negative HCV RNA at the end of the first cycle of IFN (P =.01) and by retreatment with a high IFN dose (P =. 03). Age, cirrhosis, genotype, and gamma-glutamyl transferase levels before retreatment were not significant by multivariate analysis. The excellent tolerability of IFN monotherapy retreatment makes it an option for patients who transiently cleared HCV-RNA during their first IFN course. Patients should be retreated with a high IFN dose regardless of the strength of the dose received during the previous course of treatment. PMID- 10462390 TI - Optimal treatment of biliary atresia--"halfway" there! PMID- 10462391 TI - Apoptosis and the liver: A mechanism of disease, growth regulation, and carcinogenesis. PMID- 10462392 TI - Gene therapy for hepatic fibrosis-bringing treatment into the new millennium. PMID- 10462393 TI - International Society of Paediatric Oncology, American Society of Pediatric Hematology/Oncology joint meeting and International Society of Paediatric Surgical Oncology 31st meeting. Montreal, Canada, September 13-18, 1999. Abstracts. PMID- 10462395 TI - SIOP Abstracts O-72-O-153. PMID- 10462394 TI - SIOP Abstracts O-1-O-72. PMID- 10462396 TI - SIOP Abstracts SL-1-SL-73. PMID- 10462397 TI - SIOP Abstracts GL-2-GL-26. PMID- 10462399 TI - SIOP Abstracts P-1-P-67. PMID- 10462398 TI - SIOP Abstracts HL-2-HL-3. PMID- 10462400 TI - SIOP Abstracts P-68-P-132. PMID- 10462401 TI - SIOP Abstracts P-133-P-206. PMID- 10462402 TI - SIOP Abstracts P-207-P-284. PMID- 10462403 TI - SIOP Abstracts P-285-P-375. PMID- 10462404 TI - SIOP Abstracts 1-56. PMID- 10462405 TI - SIOP Abstracts 57-112. PMID- 10462406 TI - SIOP Abstracts 113-186. PMID- 10462407 TI - IPSO Program. PMID- 10462408 TI - IPSO Abstracts. PMID- 10462409 TI - The Children's Hospitals in Montreal. PMID- 10462410 TI - G proteins, chemosensory perception, and the C. elegans genome project: An attractive story. AB - Heterotrimeric G proteins, consisting of alpha, beta, and gamma subunits, couple ligand-bound seven transmembrane domain receptors to the regulation of effector proteins and production of intracellular second messengers. G protein signaling mediates the perception of environmental cues in all higher eukaryotic organisms, including yeast, Dictyostelium, plants, and animals. The nematode Caenorhabditis elegans is the first animal to have complete descriptions of its cellular anatomy, cell lineage, neuronal wiring diagram, and genomic sequence. In a recent paper, Jansen et al. used sequence searches of the C. elegans genome database to identify all heterotrimeric G protein genes (20 Galpha, 2 Gbeta, 2 Ggamma). C. elegans encodes one ortholog of each of the four Galpha classes found in metazoans and 16 new Galpha genes. The orthologous genes are widely expressed, whereas 14 of the divergent Galpha genes are almost exclusively expressed in sensory neurons where they may regulate perception and chemotaxis. PMID- 10462411 TI - New growth factors for imaginal discs. AB - Several families of peptide growth factors are implicated in regulating cell growth and proliferation of vertebrate cells in culture. Genetic studies in Drosophila implicate some of these factors in growth control in vivo. A recent report identifies a new family of growth factors, related to chitinase enzymes, required by Drosophila imaginal disc cells in culture. It will be of interest to determine how such factors relate to size regulation during development. PMID- 10462412 TI - Limbiting outgrowth: BMPs as negative regulators in limb development. AB - Rapid progress is being made in understanding how integrated signaling pathways direct patterned outgrowth of the vertebrate limb. In contrast, the mechanisms that constrain limb outgrowth, and thus delimit adult morphology, remain poorly understood. Two recent pioneering reports have implicated bone morphogenetic proteins (BMPs) in negatively regulating the function of the apical ectodermal ridge (AER), an inductive structure required for continued proximodistal specification of limb elements. These studies provide the first insights into how the termination of a limb bud signaling center is accomplished, and intriguingly suggest how distinct aspects of limb morphogenesis are regulated. PMID- 10462413 TI - Transcriptional regulation of lymphocyte lineage commitment. AB - The development of T cells and B cells from pluripotent hematopoietic precursors occurs through a stepwise narrowing of developmental potential that ends in lineage commitment. During this process, lineage-specific genes are activated asynchronously, and lineage-inappropriate genes, although initially expressed, are asynchronously turned off. These complex gene expression events are the outcome of the changes in expression of multiple transcription factors with partially overlapping roles in early lymphocyte and myeloid cell development. Key transcription factors promoting B-cell development and candidates for this role in T-cell development are discussed in terms of their possible modes of action in fate determination. We discuss how a robust, stable, cell-type-specific gene expression pattern may be established in part by the interplay between endogenous transcription factors and signals transduced by cytokine receptors, and in part by the network of effects of particular transcription factors on each other. PMID- 10462414 TI - Controlling the urge for a Ca(2+) surge: all-or-none Ca(2+) release in neurons. AB - Changes in the intracellular calcium concentration ([Ca2+]i) convey signals that are essential to the life and death of neurons. Ca(2+)-induced Ca(2+)-release (CICR), a process in which a modest elevation in [Ca2+]i is amplified by a secondary release of Ca2+ from stores within the cell, plays a prominent role in shaping neuronal [Ca2+]i signals. When CICR becomes regenerative, an explosive increase in [Ca2+]i generates a Ca2+ wave that spreads throughout the cell. A discrete threshold controls activation of this all-or-none behavior and cellular context adjusts the threshold. Thus, the store acts as a switch that determines whether a given pattern of electrical activity will produce a local or global Ca2+ signal. This gatekeeper function seems to control some forms of Ca(2+) triggered plasticity in neurons. PMID- 10462415 TI - BMP signalling in early Xenopus development. AB - Bone morphogenetic proteins (BMPs) are typically members of the transforming growth factor beta (TGF-beta) family with diverse roles in embryonic development. At least five genes with homology to BMPs are expressed during Xenopus development, along with their receptors and intracellular signalling pathways. The evidence suggests that BMPs have roles to play in both mesoderm induction and dorsoventral patterning. Studies in Xenopus have also identified a number of inhibitory binding proteins for the classical BMPs, encoded by genes such as chordin and noggin. These proteins appear to be responsible for establishing a morphogen gradient of BMP4 activity, which specifies different dorsoventral fates in early gastrulae. An emerging theme is that inhibition of BMP signalling is an important mechanism regulating cell fate decisions in early development. PMID- 10462416 TI - Rapid structural and epigenetic changes in polyploid and aneuploid genomes. AB - Recent work with plants has demonstrated that genome instability can be triggered by a change in chromosome number arising from either whole genome duplications (polyploidy) or loss/gain of individual chromosomes (aneuploidy). This genome instability is manifested as rapid structural and epigenetic alterations that can occur somatically or meiotically within a few generations after heteroploid formation. The intrinsic instability of newly formed polyploid and aneuploid genomes has relevance for genome evolution and human carcinogenesis, and points toward recombinational and epigenetic mechanisms that sense and respond to chromosome numerical changes. PMID- 10462417 TI - Steroidogenic acute regulatory (StAR) protein: what's new? AB - In response to trophic hormone stimulation of steroidogenic adrenal and gonadal cells, the acute biosynthesis of steroid hormones occurs in the order of minutes to tens of minutes and can be contrasted to chronic regulation, which occurs on the order of hours. The steroidogenic acute regulatory (StAR) protein is an indispensable component in the acute regulatory phase and functions by rapidly mediating the transfer of the substrate for all steroid hormones, cholesterol, from the outer to the inner mitochondrial membrane where it is cleaved to pregnenolone, the first steroid formed. This transfer of cholesterol constitutes the rate-limiting step in steroidogenesis. To underscore its importance, mutations in the StAR gene have been shown to be the only cause of the potentially fatal disease lipoid congenital adrenal hyperplasia, in which affected individuals synthesize virtually no steroids. Since the cloning of the murine cDNA in 1994, many observations have substantiated the critical role of StAR in regulated steroidogenesis. The purpose of this review will be to summarize briefly some background material on StAR and then attempt to update several recent and interesting findings on the StAR protein. PMID- 10462418 TI - The nucleosome core particle: does it have structural and physiologic relevance? AB - Although the nucleosomal core particle has been extensively studied as the basic building block of chromatin, the biological significance of a unit carrying exactly 146 bp of DNA remains unclear. Herein, we present data to show that the histone octamer can stably accommodate anywhere from about 100 to 170 bp of DNA. The unfolded structures containing less than 146 bp may well be of greater biological importance than the canonical core particle. PMID- 10462419 TI - Expression profiling: DNA arrays in many guises. AB - DNA arrays have become the preferred method for large-scale expression measurement. Such data are needed in view of the large amounts of sequence data available: expression levels in a number of different tissues or situations provide a first step toward functional characterisation of new entities revealed by DNA sequencing. Although the basic principle of measurement is in all cases based on hybridisation of a mixed probe derived from tissue RNA to large sets of DNA fragments representing many genes, a number of different forms of implementation of this principle are at hand. They are briefly described and compared, emphasizing the important issue of sensitivity and sample requirements and the accessibility of the methods to academic scientists. When these factors are taken into account, it appears that, contrary to a largely prevalent impression, the "best" approach is not necessarily always provided by the widely advertised glass microarrays or oligonucleotide chips. PMID- 10462420 TI - In vitro fertilization by intracytoplasmic sperm injection. AB - Intracytoplasmic sperm injection (ICSI) is the latest, and by far the most efficient, variant of micromanipulation-assisted fertilization, whereby a single spermatozoon is selected, aspirated into a microinjection needle and injected to the oocyte cytoplasm. The development of this technique is mainly linked to application in human assisted reproduction for which it enables fertilization with defective spermatozoa that would not otherwise be able to penetrate an oocyte by their proper means. Because ICSI by-passes many steps of the natural fertilization process, it offers an extremely interesting model for the study of basic mechanisms underlying fertilization. This is particularly true for oocyte activation, whose mechanism needs to be revisited in light of the current ICSI research. The massive application of ICSI in human infertility treatment also represents a huge laboratory in which the impact of different genetic and epigenetic anomalies of the male gamete on fertilization and embryonic development can be studied. PMID- 10462421 TI - Corrigenda. PMID- 10462422 TI - Of Jewel Heritage: racial socialization and racial identity attitudes amongst adolescents of mixed African-Caribbean/White parentage. AB - Mixed parentage adolescents form an increasing proportion of ethnic minority adolescents in Britain. Few studies have investigated their views and perceptions of their racial identity in terms of their Black heritage. This exploratory study investigated their attitudes towards Blackness, and examined the relationship that this might have with their reported experiences of racial socialization as well as with their self-esteem. Older adolescents were more likely to have positive racial identity attitudes, and a positive relationship was found between racial identity attitudes and self-esteem. Reported frequency of certain types of racial socialization messages increased with age. This was especially pronounced with messages relating to the development of racial pride and messages relating to issues around racism. Those adolescents who lived with both parents reported receiving more proactive racial socialization messages. Tentative conclusions include the possibility that dual identification and multicultural environments are associated with a positive sense of racial identity. These issues need to be investigated further. PMID- 10462423 TI - IQ scores among homeless older adolescents: characteristics of intellectual performance and associations with psychosocial functioning. AB - Intellectual performance and the associations of IQ with the quality of psychosocial functioning were studied in a sample of homeless older adolescents. Fifty homeless older adolescents (ages 16-21) completed the Wechsler Adult Intelligence Scale-Revised (WAIS-R) and a questionnaire assessing psychosocial functioning and sexual risk factors. The WAIS-R scores were comparable to population means, with performance IQ scores tending to be higher than verbal IQ scores. The IQ was unrelated to the duration of homelessness. Higher IQ scores were significantly correlated with only a minority of the measures of psychosocial functioning, including less self-reported depression and lower reported delinquency, but also less self-control in high-risk sexual situations, less perceived peer support for safer sex, and a higher perceived likelihood of acquiring HIV. PMID- 10462424 TI - Adolescent children of Vietnam combat veteran fathers: a population at risk. AB - This exploratory study compared 28 adolescent children of Vietnam combat veterans with 28 adolescents whose fathers were not in Vietnam on multiple variables related to (1) social and personal adjustment, (2) attitudes toward parents, and (3) personality development. The majority of outcomes were not significantly different. The children of combat veterans, however, showed poorer attitudes toward school; more negative attitudes toward their fathers; elevated scores of depression, tension, apprehension, and anxiety; lowered scores on creativity; and their mothers rated their behaviors as more problematic. Interactions with gender were not significantly different. PMID- 10462425 TI - The effects of family structure and SES on family processes in early adolescence. AB - This study investigates parent-adolescent relationships in early adolescence, focusing on the effects of dyadic communication, family cohesion, family structure, and SES on family conflict in three ethnic groups: African-American (n =1886), Mexican-American (n =2657), and Euro-American (n =3052). Sixth, seventh and eighth grade girls and boys completed questionnaires and results were analysed using independent samples t -tests and multiple regression. Results showed 3-year increases in family conflict in all three ethnic groups. There were significant differences in scale scores on conflict, communication, and family cohesion between nuclear and single-parent families. Some ethnic and gender differences were also found. Discussion focuses on the implications of the findings for family life. PMID- 10462426 TI - The effects of partner insistence of condom usage on perceptions of the partner, the relationship, and the experience. AB - The present study examined the effects of an individual insisting on condom usage on how he or she is viewed by their partner. Participants were led through a realistic role-play scenario in which, after dating a new partner for "a while", they engaged in sexual intercourse. Results were such that after first time sex, participants indicated feeling more responsible, less at-risk, and less worried when a condom was used than when no condom was used. Participants evaluated their partner as more responsible, more caring, and less likely to have a sexually transmitted disease when a condom was used. Further, when a condom was used, the relationship was evaluated as enhanced, closer, more intimate, and more likely to be long lasting. The insistence of condom use by the participant's partner also resulted in less guilt and regret regarding the sexual experience itself. These results generally held true for both male and female participants. PMID- 10462427 TI - Repetition of deliberate self-harm by adolescents: the role of psychological factors. AB - The aim of this study was to examine the relationship between psychological variables and repetition of deliberate self-harm by adolescents (n=45) aged 13-18 years who had been admitted to a general hospital having taken overdoses. Standardized measures of depression, hopelessness, suicidal intent, impulsivity, trait and state anger, self-esteem and problem-solving (both self-report and observer-rated) were administered to the adolescents while still in the general hospital. Repetition was assessed on the basis of previous overdoses (n=14) and repetition of self-harm (self-poisoning and self-injury) during the subsequent year (n=9). Adolescents with a history of a previous overdose and/or who repeated self-harm during the following year (n=18) differed from non-repeaters in having higher scores for depression, hopelessness and trait anger, and lower scores for self-esteem, self-rated problem-solving and effectiveness of problem-solving rated on the basis of the Means End Problem Solving test, all measured at the initial assessment. These differences largely disappeared when level of depression was controlled for. Similarly, differences found between repeaters and non-repeaters in the year following the index overdoses for problem-solving were much reduced when account was taken of differences in depression scores. Depression is a key factor associated with risk of repetition of adolescent self harm (and hence of suicide risk). In the management of adolescents who have harmed themselves, careful assessment of depression and appropriate management of those who are depressed is essential. PMID- 10462428 TI - Eating disorder behaviors of ethnically diverse urban female adolescent athletes and non-athletes. AB - This study compared Caucasian, Hispanic, and African-American urban adolescent athlete and non-athlete females for relative frequency of behavioral and psychological indices of eating disorders, while controlling for physical size. High school female athletes (n=571) and non-athletes (n=463) completed the Eating Disorder Inventory (EDI) composed of eight subscales that measure behavioral and psychological indices common in anorexia nervosa and bulimia nervosa. The MANCOVA for the main effect of ethnicity showed Caucasians and Hispanics scored significantly higher than African-Americans on six of the eight behavioral and psychological subscales of the EDI (p<0. 05). A MANCOVA for the main effect of athletic status showed no significant differences between athletes and non athletes on the behavioral subscales of the EDI (p>0.05). This study suggests that Hispanic and Caucasian urban adolescent females are comparably more at-risk for eating disorders than African-American urban adolescent females. In addition, athletes were no more at-risk than non-athletes for eating disordered behaviors. PMID- 10462429 TI - Subclinical eating disorders in adolescent women: a test of the continuity hypothesis and its psychological correlates. AB - Subclinical eating disorders are common in adolescent women. In this study the severity of disturbed eating and its psychological correlates were examined in 207 college freshmen. The results indicated that 9% fell into the probable bulimic or dieter at-risk categories, 23% were classified as intensive dieters, 17% as casual dieters, and over half (51%) of the sample was classified as non dieters. Depression, dysfunctional thinking, and disturbed eating attitudes were found to correlate with the severity of eating pathology. Support was found for the continuity hypothesis of eating disorders. PMID- 10462430 TI - Does self-competence predict gender differences in adolescent depression and anxiety? AB - This longitudinal study examined 75 young adolescents to explore whether self competence predicts the emergence of gender differences in depression and anxiety. During both 6th and 7th grade, boys reported significantly higher levels of self-competence than did girls. In addition, boys were significantly less depressed and anxious than girls in 7th grade, but not in 6th grade. Finally, when the variance contributed by self-competence was accounted for, the relationship between gender and trait anxiety weakened and the relationship between gender and depression became non-significant. These results support the hypothesis that self-competence is partially responsible for the emergence of gender differences in depression and anxiety during early adolescence. PMID- 10462431 TI - Brief report. The overdose process-adolescents' experience of taking an overdose and their contact with services. PMID- 10462432 TI - Brief report. The psychometric assessment of anger in male and female adolescents resident at a secure youth treatment centre. PMID- 10462433 TI - Brief report: types of bullying among Italian school children. AB - The main aim of this research was to investigate the prevalence of bullies and victims and the types and places of bullying. A questionnaire on bullying was completed by 113 girls and 125 boys in a middle school in Rome (aged 11-14 years). Over half of all students had bullied others in the previous 3 months, and nearly half had been bullied. Boys bullied more than girls, and both boys and girls tended to be bullied by boys. Boys were more likely to suffer direct bullying such as being threatened or physically hurt. Most of the bullying took place in the classroom; boys were likely to be bullied in the toilet. PMID- 10462434 TI - Call for papers PMID- 10462435 TI - Cytochrome P450 and the individuality of species. AB - The P450 superfamily is expanding rapidly on many fronts. Arabidopsis genomic sequencing is producing about 2 to 3 novel P450s per week, with some clusters containing 9-14 genes. Bacterial genomes also carry surprises, such as the 20 P450s found in Mycobacterium tuberculosis and the 7 in Bacillus subtilis. The race to finish the human genome has already identified the majority of human P450s, some by expressed sequence tags only. The rapid discovery of new genes is being complemented by detailed analysis of our human genes to identify and characterize the complete set of human P450 polymorphisms and disease-causing mutations, one aspect of our "chemical individuality." Phylogenetic trees are included for plant, fungal, animal, and bacterial P450s. Emphasis is given to the higher order nomenclature of P450 clans, as a tool to see the larger picture of P450 evolution. Arabidopsis is the current record holder in P450 genes, with 186 named genes and a prediction of 350 in the total genome to be completed next year. The biosynthesis of cholesterol in bacteria is discussed in relation to CYP51 as a lanosterol 14 alpha-demethylase. This enzyme may have been the first eukaryotic P450. PMID- 10462436 TI - P450 gene induction by structurally diverse xenochemicals: central role of nuclear receptors CAR, PXR, and PPAR. AB - The biochemistry of foreign compound metabolism and the roles played by individual cytochrome P450 (CYP) enzymes in drug metabolism and in the toxification and detoxification of xenochemicals prevalent in the environment are important areas of molecular pharmacology and toxicology that have been widely studied over the past decade. Important advances in our understanding of the mechanisms through which foreign chemicals impact on these P450-dependent metabolic processes have been made during the past 2 years with several key discoveries relating to the mechanisms through which xenochemicals induce the expression of hepatic P450 enzymes. Roles for three "orphan" nuclear receptor superfamily members, designated CAR, PXR, and PPAR, in respectively mediating the induction of hepatic P450s belonging to families CYP2, CYP3, and CYP4 in response to the prototypical inducers phenobarbital (CAR), pregnenolone 16alpha carbonitrile and rifampicin (PXR), and clofibric acid (PPAR) have now been established. Two other nuclear receptors, designated LXR and FXR, which are respectively activated by oxysterols and bile acids, also play a role in liver P450 expression, in this case regulation of P450 cholesterol 7alpha-hydroxylase, a key enzyme of bile acid biosynthesis. All five P450-regulatory nuclear receptors belong to the same nuclear receptor gene family (family NR1), share a common heterodimerization partner, retinoid X-receptor (RXR), and are subject to cross-talk interactions with other nuclear receptors and with a broad range of other intracellular signaling pathways, including those activated by certain cytokines and growth factors. Endogenous ligands of each of those nuclear receptors have been identified and physiological receptor functions are emerging, leading to the proposal that these receptors may primarily serve to modulate hepatic P450 activity in response to endogenous dietary or hormonal stimuli. Accordingly, P450 induction by xenobiotics may in some cases lead to a perturbation of endogenous regulatory circuits with associated pathophysiological consequences. PMID- 10462437 TI - How similar are P450s and what can their differences teach us? AB - Cytochromes P450 form a very large superfamily of proteins which metabolize substrates from steroids to fatty acids to drugs and are found in organisms from protists to mammals. P450s all appear to take on a similar structural fold, yet frequently having less than 20% sequence identity and having vastly different substrates. Within the structural fold there appears to be a highly conserved core, as determined from the comparison of the structures of the six crystallized, soluble P450s. There are also variable regions which by and large appear to be associated with substrate recognition, substrate binding, and redox partner binding. Molecular dynamics simulations of motion in P450cam and P450BM-3 indicate that substrate binding and product release require substantial motion around the "substrate access channel." Additionally, at the 11th International Conference on Cytochrome P450 Biochemistry, Biophysics, and Molecular Biology and briefly here, the first structure of a microsomal eukaryotic P450 will be presented and compared to the already determined structures by Drs. Johnson and McRee. Finally, with a better understanding of the structure/function relationship of P450s, one will be better able to modify P450s to metabolize the substrates of choice or produce needed valuable chemicals. PMID- 10462438 TI - Prediction of the active-site structure and NAD(+) binding in SQD1, a protein essential for sulfolipid biosynthesis in Arabidopsis. AB - Sulfolipids of photosynthetic bacteria and plants are characterized by their unique sulfoquinovose headgroup, a derivative of glucose in which the 6-hydroxyl group is replaced by a sulfonate group. These sulfolipids have been discussed as promising anti-tumor and anti-HIV therapeutics based on their inhibition of DNA polymerase and reverse transcriptase. To study sulfolipid biosynthesis, in particular the formation of UDP-sulfoquinovose, we have combined computational modeling with biochemical methods. A database search was performed employing the derived amino acid sequence from SQD1, a gene involved in sulfolipid biosynthesis of Arabidopsis thaliana. This sequence shows high similarity to other sulfolipid biosynthetic proteins of different organisms and also to sugar nucleotide modifying enzymes, including UDP-glucose epimerase and dTDP-glucose dehydratase. Additional biochemical data on the purified SQD1 protein suggest that it is involved in the formation of UDP-sulfoquinovose, the first step of sulfolipid biosynthesis. To understand which aspects of epimerase catalysis may be shared by SQD1, we built a three-dimensional model of SQD1 using the 1.8 A crystallographic structure of UDP-glucose 4-epimerase as a template. This model predicted an NAD(+) binding site, and the binding of NAD(+) was subsequently confirmed by enzymatic assay and mass spectrometry. The active-site interactions together with biochemical data provide the basis for proposing a reaction mechanism for UDP sulfoquinovose formation. PMID- 10462439 TI - A novel proanthocyanidin IH636 grape seed extract increases in vivo Bcl-XL expression and prevents acetaminophen-induced programmed and unprogrammed cell death in mouse liver. AB - Several molecular events in the apoptotic or necrotic death of hepatocytes induced by acetaminophen (AAP) now appear to be well defined. Recent studies also indicate that select expression of bcl-Xl is possibly modified during AAP-induced liver injury. The purpose of this study was several-fold: (i) to examine the hepatoprotective ability of short-term (3-day) and long-term (7-day) exposures of a grape seed proanthocyanidin extract (GSPE) on AAP-induced liver injury and animal lethality; (ii) to monitor effects of GSPE on one of the prime targets of AAP, i.e., hepatocellular genomic DNA and associated apoptotic and necrotic death; and (iii) to unravel changes in the pattern of expression of an antiapoptotic gene, bcl-Xl in the liver. In order to investigate these events, male ICR mice (30-40 g) were administered nontoxic doses of GSPE (3 or 7 days, 100 mg/kg, po), followed by hepatotoxic doses of AAP (400 and 500 mg/kg, ip), and sacrificed 24 h later. Serum was analyzed for alanine aminotransferase activity (ALT) and the liver for histopathological diagnosis of apoptosis/necrosis. The ability of AAP to promote apoptotic DNA fragmentation and its counteraction by GSPE in the liver was also evaluated quantitatively (by a sedimentation assay) and qualitatively (by agarose gel electrophoresis). Portions of livers were also subjected to Western blot analysis (27,000g fraction of liver homogenates) to examine the pattern of expression of cell death inhibitory gene bcl-Xl. Results indicate that 7-day GSPE preexposure induced dramatic protection and markedly decreased liver injury and animal lethality culminated by AAP, when compared to a short-term 3-day exposure. Abrogation of toxicity was also mirrored in DNA fragmentation. Histopathological evaluation of liver sections showed remarkable counteraction of AAP-toxicity by this novel GSPE and substantial inhibition of both apoptotic and necrotic liver cell death. Agarose gel electrophoresis revealed that 7-day GSPE preexposure prior to AAP administration completely blocked Ca(2+)/Mg(2+)-Ca(2+)/Mg(2+)-dependent-endonuclease-mediated ladder-like fragmentation of genomic DNA and significantly altered the bcl-Xl expression. The most dramatic changes observed in this study were: (i) substantial increase in the expression of bcl-Xl in the liver by 7-day GSPE exposure alone; (ii) significant modification bcl-Xl expression by AAP alone; and (iii) dramatic inhibition of AAP-induced modification of bcl-Xl (phosphorylation?) expression by GSPE. In summary, these observations demonstrate that GSPE preexposure may significantly attenuate AAP-induced hepatic DNA damage, apoptotic and necrotic cell death of liver cells, and, most remarkably, antagonize the influence of AAP induced changes in bcl-Xl expression in vivo. PMID- 10462440 TI - Effects of chlorpromazine on actin polymerization: slackening of filament elongation and filament annealing. AB - We have analyzed the effect of chlorpromazine (CPZ) on pure actin. We have found that CPZ quenches Trp-79 and Trp-86 fluorescence and, in agreement with an earlier report on conventional actin, inhibits actin polymerization, lowering the extent of polymerization. Moreover, novel polymerization data are presented indicating that CPZ decreases the maximum polymerization rate in a dose-dependent manner. The assembly inhibition results from the slackening of oligomer formation during the early stages of polymerisation, of filament elongation and of filament annealing. Finally, CPZ strongly inhibits actin filament network formation. PMID- 10462441 TI - Topoisomerase II is a cellular target for antiproliferative cobalt salicylaldoxime complex. AB - Topoisomerase II is a cellular target for a number of clinically relevant antitumor drugs. To elucidate the possible cellular target for the antiproliferation activity of cobalt salicylaldoxime (CoSAL), which inhibits 50% of leukemic cell proliferation at a concentration of 60 microM, DNA binding studies and studies of the action of this complex on topoisomerase II catalytic activities were carried out. The results from DNA binding studies show that CoSAL binds DNA strongly with a stoichiometric ratio of two drug molecules for five nucleotide bases and shows a mode of interaction similar to that of DNA groove binding agents. The results from topoisomerase II inhibition studies show that the complex inhibits the relaxation activity of topoisomerase II in a dose dependent manner and poisons its activity through cleavage complex formation. To see if the hydroxyl group present on imine nitrogen is involved in topoisomerase II poisoning, we synthesized an analogue of CoSAL in which the hydroxyl group was replaced with semicarbazone. This complex too binds DNA with an affinity similar to that of CoSAL, but with a small difference in the mode of interaction; however, it marginally inhibits leukemic cell proliferation and does not inhibit topoisomerase II activity, which suggests the involvement of a hydroxyl group. An immunoprecipitation assay was conducted which showed that the cleavage complex formed in the presence of CoSAL contained 75% of the complex, while the other complex shows only 7. 65%. Cyclic voltametric spectra of the complexes in the presence of DNA show that they do not oxidize DNA. These results suggest that CoSAL shows a bidirectional mode of interaction with enzyme and DNA and inhibits topoisomerase II activity by forming a drug-mediated cleavage complex. Our data strongly suggest that topoisomerase II may be one of the cellular targets for antiproliferation activity of CoSAL. PMID- 10462442 TI - Nuclear magnetic resonance and oxygen affinity study of cesium binding in human erythrocytes. AB - We investigated the interaction of the cesium ion (Cs(+)) with the anionic intracellular components of human red blood cells (RBCs); the components studied included 2,3-bisphosphoglycerate (BPG), ADP, ATP, inorganic phosphate (P(i)), carbonmonoxy hemoglobin (COHb), and RBC membranes. We used spin-lattice (T(1)) and spin-spin (T(2)) (133)Cs NMR relaxation measurements to probe Cs(+) binding, and we found that Cs(+) bound more strongly to binding sites in BPG and in RBC membranes than in any other intracellular component in RBCs at physiologic concentrations. By using James-Noggle plots, we obtained Cs(+) binding constants per binding site in BPG (66 +/- 8 M(-1)), ADP (19 +/- 1 M(-1)), ATP (25 +/- 3 M( 1)), and RBC membranes (55 +/- 2 M(-1)) from the observed T(1) values. We also studied the effect of Cs(+) on the oxygen (O(2)) affinity of purified Hb and of Hb in intact RBCs in the absence and in the presence of BPG. In the absence of BPG, the O(2) affinity of Hb decreased upon addition of Cs(+). However, in the presence of BPG, the O(2) affinity of Hb increased upon addition of Cs(+). The O(2) affinity of Cs(+)-loaded human RBCs was larger than that of Cs(+)-free cells at the same BPG level. (31)P NMR studies on the pH dependence of the interaction between BPG and Hb indicated that the presence of Cs(+) resulted in a smaller fraction of BPG available to bind to the cleft of deoxyHb. Our NMR and O(2) affinity data indicate that a strong binding site for Cs(+) in human RBCs is BPG. A partial mechanism for Cs(+) toxicity might arise from competition between Cs(+) and deoxyHb for BPG, thereby increasing oxygenation of Hb in RBCs, and thus decreasing the ability of RBCs to give up oxygen in tissues. The presence of Cs(+) at 12.5 mM in intact human RBCs containing BPG at normal concentrations did not, however, alter significantly the O(2) affinity of Hb, thus ruling out the possibility of Cs(+)-BPG interactions accounting for Cs(+) toxicity in this cell type. PMID- 10462443 TI - Different regulation of the expression of mouse hepatic cytochrome P450 2B enzymes by glucocorticoid and phenobarbital. AB - The effect of the synthetic glucocorticoid, dexamethasone, and phenobarbital upon the expression of Cyp2b9 and Cyp2b10, major CYP2B subfamilies in the mouse, was differentiated in C57BL/6 mouse liver and hepatocytes in primary culture. Overall expression was higher in the untreated female liver than in the male liver. More Cyp2b9 than Cyp2b10 mRNA was present in the female liver, whereas the level of Cyp2b10 was higher in the male. Phenobarbital increased Cyp2b10 expression more than did Cyp2b9 in both sexes. Treatment with dexamethasone markedly induced Cyp2b10 expression dose dependently, but simultaneously suppressed Cyp2b9 in both sexes. Evidence of this was obtained both in vivo and in hepatocyte culture. Furthermore, the existence of at least two unknown species of CYP2B, whose expressions were either increased or decreased by dexamethasone was suggested. Adrenalectomy increased the expression of Cyp2b9 and Cyp2b10 mRNAs, especially that of Cyp2b9 in the male liver. In addition, the expression of one unknown species which was constitutively suppressed increased in adrenalectomized male mice. That the treatment of dexamethasone or adrenalectomy altered the expression of CYP2B subfamilies suggests that endogenous glucocorticoid hormone plays a basic role in the constitutive expression of cytochrome P450. Furthermore, the sex-related difference in the expression of Cyp2b9 and Cyp2b10 suggests that sex dependent secretion of endogeneous modulating factors is involved in the regulatory pathway. PMID- 10462444 TI - Multiple unfolded states of glutathione transferase bbGSTP1-1 by guanidinium chloride. AB - Inactivation, dissociation, and unfolding of the homodimeric glutathione transferase (bbGSTP1-1) from Bufo bufo embryos were investigated at equilibrium, using guanidinium chloride (GdmCl) as denaturant. Protein transitions were monitored by enzyme activity, intrinsic fluorescence, far UV circular dichroism, glutaraldehyde cross-linking, and gel-filtration chromatography. At low denaturant concentrations (less than 0.5 M), reversible inactivation of the enzyme occurs. At denaturant concentrations between 0.5 and 1.5 M the enzyme progressively dissociates into structured monomers. At higher denaturant concentrations the monomers unfold completely. Refolding studies indicate that a total reactivation occurs only by starting from the enzyme denatured at concentrations below 0.5 M. The enzyme denatured at GdmCl concentrations higher than 0.5 M only partially refolds. Globally our results indicate that unfolding of the amphibian bbGSTP1-1 is a multistep process, i.e., inactivation of the structured dimer, dissociation into partially structured monomers, followed by complete unfolding. PMID- 10462445 TI - Targeting of venom phospholipases: the strongly anticoagulant phospholipase A(2) from Naja nigricollis venom binds to coagulation factor Xa to inhibit the prothrombinase complex. AB - The strongly anticoagulant basic phospholipase A(2) (CM-IV) from Naja nigricollis venom has previously been shown to inhibit the prothrombinase complex of the coagulation cascade by a novel nonenzymatic mechanism (S. Stefansson, R. M. Kini, and H. J. Evans Biochemistry 29, 7742-7746, 1990). That work indicated that CM-IV is a noncompetitive inhibitor and thus it interacts with either factor Va or factor Xa, or both. We further examined the interaction of CM-IV and the protein components of the prothrombinase complex. Isothermal calorimetry studies indicate that CM-IV does not bind to prothrombin or factor Va, but only to factor Xa. CM IV has no effect on the cleavage of prothrombin by factor Xa in the absence of factor Va. However, in the presence of factor Va, CM-IV inhibits thrombin formation by factor Xa. With a constant amount of CM-IV, raising the concentration of factor Va relieved the inhibition. The phospholipase A(2) enzyme inhibits by competing with factor Va for binding to factor Xa and thus prevents formation of the normal Xa-Va complex or replaces bound factor Va from the complex. Thus factor Xa is the target protein of this anticoagulant phospholipase A(2), which exerts its anticoagulant effect by protein-protein rather than protein-phospholipid interactions. PMID- 10462446 TI - Muscarinic acetylcholine receptor (mAChR) inhibitor from snake venom: interaction with subtypes of human mAChR. AB - Snake venoms can contain a variety of well-studied neurotoxins, especially nicotinic acetylcholine receptor inhibitor, normally called postsynaptic neurotoxin. Karlsson first reported muscarinic acetylcholine receptor (mAChR) inhibitor from snake venom. In a previous study in our laboratory, we found a mAChR inhibitor from Naja naja sputatrix venom that bound to rat brain synaptosomes. Brain synaptosomes contain all subtypes of mAChRs, and thus the exact selectivity of the inhibitor could not be determined. mAChR inhibitor from N. naja sputatrix venom was purified and the binding to all human mAChR subtypes (M1, M2, M3, M4, and M5) was investigated and is reported in this communication. The inhibitor bound to all subtypes of the human mAChR, but showed considerably high selectivity for the M5 subtype. It was also found that the reduction of disulfide bonds in the inhibitor eliminated the binding to the mAChR. This suggests that a specific tertiary conformation maintained by disulfide bonds is essential for binding to the mAChR. An oligo peptide, QIHDNCYNE, comparable to a part of the inhibitor molecule, was synthesized and studied for its binding to the mAChR. The synthetic peptide did not show any binding activity, suggesting this portion of the inhibitor molecule is not involved in mAChR binding. The selective binding of the M5 mAChR subtype to antagonists has not yet been reported. Therefore, the purified inhibitor reported in this communication may be a useful tool to clarify the mechanism of muscarinic cholinergic transmission. PMID- 10462447 TI - Kinetic characterization of mitochondrial complex I inhibitors using annonaceous acetogenins. AB - The NADH:ubiquinone oxidoreductase (complex I) of the mitochondrial respiratory chain is by far the largest and most complicated of the proton-translocating enzymes involved in the oxidative phosphorylation. Many clues regarding the electron pathways from matrix NADH to membrane ubiquinone and the links of this process with the translocation of protons are highly controversial. Different types of inhibitors become valuable tools to dissect the electron and proton pathways of this complex enzyme. Therefore, further knowledge of the mode of action of complex I inhibitors is needed to understand the underlying mechanism of energy conservation. This study presents for the first time a detailed exploration of the inhibitory action of the Annonaceous acetogenins, the most powerful inhibitors of the mammalian enzyme, taking as the head-series rolliniastatin-1, rolliniastatin-2, and corossolin. Despite their close chemical resemblance, each of them inhibits the complex I with different kinetic features reflecting differential binding to the enzyme. PMID- 10462448 TI - Incorporation of 1-[1-(13)C]Deoxy-D-xylulose in chamomile sesquiterpenes. AB - Incorporation of synthetically prepared 1-[1-(13)C]deoxy-d-xylulose into chamomile sesquiterpenes has been achieved by injecting an aqueous solution into the anthodia of the plant. The analysis of labeling patterns and absolute (13)C abundances of the isolated sesquiterpenes bisabololoxide A (1), bisabololoxide B (2), and chamazulene (3) using quantitative (13)C NMR spectroscopy showed that 1 [1-(13)C]deoxy-d-xylulose was efficiently incorporated in all three isoprene building blocks of the sesquiterpenes. A significantly lower (13)C abundance of the labeled carbon atom in the biogenetically terminal isoprene unit confirms the mixed biosynthesis of this unit, involving both the mevalonic acid pathway and the methylerythritol phosphate pathway. PMID- 10462449 TI - Human selenium-dependent thioredoxin reductase from HeLa cells: properties of forms with differing heparin affinities. AB - The TrxRl form of thioredoxin reductase (TrxR) was the major form of the enzyme isolated from HeLa cells grown in a fermentor at 35 degrees C under controlled aeration conditions favorable to growth, nominally 30% of saturation of dissolved oxygen or 8 ml of oxygen in a liter of medium. This TrxR1 form was not retained on a heparin affinity matrix, it contained one selenium per subunit, was highly active catalytically, and showed strong cross-reactivity with anti-rat liver TrxR1 polyclonal antibodies. At higher aeration, 50% of saturation of dissolved oxygen or 12 ml of oxygen in a liter of medium, HeLa cell growth was slower and additional TrxR forms that bound to heparin were present in purified enzyme preparations. A minor component, TrxR2, the mitochondrial form of TrxR, was detected in the heparin-bound enzyme fraction. One enzyme form that contained less selenium (ca. 0.5 Se per TrxR subunit) was only about 50% as active with thioredoxin or 5,5'dithiobis(2-nitrobenzoic acid) as substrate. Cross-reactivity of this form with anti-rat liver TrxR1 polyclonal antibodies was very weak. The isoelectric point of the low Se enzyme, 5.85, was higher than that, 5.2-5.4, of normal Se content enzyme. Affinity of purified fully active TrxR1 to heparin could be induced by reduction with NADPH or tris-(2-carboxyethyl)phosphine (TCEP). Under anaerobic conditions there was complete retention of Se indicating that an enzyme conformation change effected by reduction was involved. The TCEP reduced enzyme form was very oxygen labile and upon exposure to air both the Se content and catalytic activity decreased by about 50%. Addition of millimolar concentrations of NADPH or NADP(+) to the TCEP-reduced enzyme gave full protection from oxygen inactivation. TrxR1 exhibited weak peroxidase activity with H(2)O(2) as substrate in the presence of an NADPH-generating system but this activity was unstable. Specific alkylation of the selenocysteine residue of TrxR1 which completely inhibits the NADPH-dependent reduction of disulfides also destroyed peroxidase activity. PMID- 10462450 TI - Amino acid sequences of two high-potential iron-sulfur proteins (HiPIPs) from the moderately halophilic purple phototrophic bacterium, Rhodospirillum salinarum. AB - The amino acid sequences of two very different high-potential iron-sulfur protein (HiPIP) isozymes have been determined from the moderately halophilic purple phototrophic bacterium, Rhodospirillum salinarum. Iso-1 HiPIP, which is monomeric and contains 57 amino acid residues, is most similar to the Thiobacillus ferrooxidans iron-oxidizing enzyme (45% identity and a 6-residue deletion). On the other hand, iso-2 HiPIP, which is isolated as an oligomer, contains a peptide chain with 54 amino acid residues. It is the smallest reported to date and is only 31% identical to iso-1 HiPIP. A massive deletion of 17 residues is found at the N-terminus, such that only 2 residues remain prior to the first cysteine. Iso 2 HiPIP also has a 12-residue insertion and a 5-residue deletion. Prior to this study, there were only 2 absolutely conserved residues (Tyr 19 and Gly 75, Chromatium numbering) in addition to the 4 iron-sulfur cluster binding cysteine residues among the 13 HiPIPs sequenced to date. We found that Tyr 19 is absent in iso-2 HiPIP along with the entire N-terminal loop. Moreover, Gly 75 is substituted in both R. salinarum HiPIPs. These characteristics make the R. salinarum HiPIPs, and especially iso-2, the most divergent yet characterized. PMID- 10462451 TI - Functional organization and evolution of mammalian hexokinases: mutations that caused the loss of catalytic activity in N-terminal halves of type I and type III isozymes. AB - Mammalian hexokinases are believed to have evolved from a 100-kDa hexokinase which itself is a product of duplication and fusion of an ancestral gene encoding a 50-kDa glucose 6-phosphate-sensitive hexokinase. Type II hexokinase has been shown to possess two distinct functional active sites, one in each half, which functionally resemble the original 100-kDa hexokinase, whereas type I and III isozymes possess only one active site in the C-terminal halves. This study was conducted to identify which mutations caused the loss of catalytic activity in the N-terminal halves of type I and III isozymes. Arg 174 and Ser 447 in type I isozyme and Asp 244 in type III isozyme are speculated to be the cause, because they reside adjacent to the "catalytic" site and corresponding residues, Gly 174, Asp 447, and Gly 231, are conserved in the N-terminal half of type II isozyme as well as all other 50-kDa units that possess catalytic activity. Mutations G174R and D447S in the N-terminal half of type II isozyme reduced specific activity by approximately 79 and 57%, respectively. Therefore, neither mutation alone can account for the inactivation of the N-terminal active site in type I isozyme. Either mutation, G174R or D447S, had moderate effects on Michaelis constants, K(m), for glucose and ATP. Mg(2+). Intriguingly, mutation D447S introduced a novel inhibition by unchelated ATP (K(i) = 68 microM ATP, competitive vs ATP. Mg(2+)) to the N-terminal active site of type II isozyme. Mutation G231D caused instability to type II hexokinase and near complete loss of catalytic activity (95%), suggesting that mutation G231D not only hinders catalysis at the N terminal active site but also leads to structural instability in type II hexokinase. PMID- 10462452 TI - N-Ethylmaleimide-modified Hsp70 inhibits protein folding. AB - Hsp70 molecular chaperones facilitate protein folding and translocation by binding to hydrophobic regions of nascent or unfolded proteins, thereby preventing their aggregation. N-Ethylmaleimide (NEM) inhibits the ATPase and protein translocation-stimulating activities of the yeast Hsp70 Ssa1p by modifying its three cysteine residues, which are located in its ATPase domain. NEM alters the conformation of Ssa1p and disrupts the coupling between its nucleotide- and polypeptide-binding domains. Ssa1p and the yeast DnaJ homolog Ydj1p constitute a protein folding machinery of the yeast cytosol. Using firefly luciferase as a model protein to study chaperone-dependent protein refolding, we have found that NEM also inhibits the protein folding activity of Ssa1p. Interestingly, the NEM-modified protein (NEM-Ssa1p) is a potent inhibitor of protein folding. NEM-Ssa1p can prevent the aggregation of luciferase and stimulate the ATPase activity of Ssa1p suggesting that it acts as an inhibitor by binding to nonnative forms of luciferase and by competing with them for the polypeptide binding site of Ssa1p. NEM-Ssa1p inhibits Ssa1p/Ydj1p-dependent protein refolding at different stages indicating that the chaperones bind and release nonnative forms of luciferase multiple times before folding is completed. PMID- 10462453 TI - Inactivation of glucose oxidase by diperoxovanadate-derived oxidants. AB - Inactivation of glucose oxidase occurred in the presence of bromide, vanadate, H(2)O(2), and phosphate (the bromide system), and this was prevented by NADH or phenol red, a bromine acceptor. Glucose oxidase present during the reaction between diperoxovanadate and a reduced form of vanadate, vanadyl (the vanadyl system), but not added after mixing the reactants, was inactivated, and this was accompanied by a loss of binding of the dye, Coomassie blue, to the protein. The transient intermediate of the type OVOOV(O(2)), known to form in these reactions and used in the oxidation of bromide ion and NADH, appears to be responsible for inactivating glucose oxidase. In both systems, the inactivation of the enzyme was prevented by histidine and DTT, known to quench singlet-oxygen. By direct measurement of 1270-nm emission of singlet-oxygen, its generation was demonstrated in the bromide system, and in the reaction of hypohalous acids with diperoxovanadate, but not in the vanadyl system. By themselves both hypohalous acids, HOCl and HOBr inactivated glucose oxidase, and their prior reaction with H(2)O(2) during which singlet-oxygen was released, protected the enzyme. The results provide support for possible oxidative inactivation of glucose oxidase by diperoxovanadate-derived oxidants. PMID- 10462454 TI - The functional role of a DNA primase in chloroplast DNA replication in Chlamydomonas reinhardtii. AB - A complementation experiment was developed to identify the protein component that is essential for the in vitro replication of a cloned template containing a chloroplast DNA replication origin of Chlamydomonas reinhardtii. Using this method, we have identified a DNA primase activity that copurified with DNA polymerase from the crude protein mixture. The primase catalyzed the synthesis of short RNA primers on single-stranded DNA templates. Among the synthetic templates, the order of preference was poly(dA), poly(dT), and poly(dC). The primer size range for these templates was 11-18, 5-12, and 3-11 nucleotides, respectively. On a single-stranded template containing the chloroplast DNA replication origin, the primer length range reached 19 to 27 nucleotides, indicating a better processtivity. Several initiation sites were mapped on both strands of the cloned replication origin. Some preferential initiation sites were located on A tracks spaced at one helical turn apart within the bending locus. Primase improved the template specificity of the in vitro DNA replication system and enhanced the incorporation of radioactive dATP into the supercoiled template containing the core sequences of the chloroplast DNA replication origin. PMID- 10462455 TI - Purification, properties, and amino acid sequence of a hemoglobinuria-inducing phospholipase A(2), MiPLA-1, from Micropechis ikaheka venom. AB - Dark-colored urine is one of the clinical symptoms of envenomation by Micropechis ikaheka (New Guinea small-eyed snake). We have purified a phospholipase A(2), MiPLA-1, which induces dark-colored urine in experimental mice, to homogeneity. The analysis of the dark-colored urine by electrophoresis and N-terminal sequence determination indicated that the color of mouse urine is due to hemoglobin in the urine but not myoglobin. MiPLA-1 is the first hemoglobinuria-inducing toxin. Insignificant hemolytic activity of MiPLA-1 indicates that hemoglobinuria is not due to lysis of erythrocytes by MiPLA-1. This suggests that hemoglobinuria induced by MiPLA-1 may be due to kidney leakage caused by unknown mechanisms. MiPLA-1 also showed other biological effects, including myotoxicity as well as anticoagulant and antiplatelet effects. Structural studies show that MiPLA-1 is a basic protein with a molecular mass of 14041.60 +/- 1.78 as determined by electrospray mass spectrometry. We have determined the complete amino acid sequence of MiPLA-1. It is a 124-amino-acid protein with a "pancreatic loop" and belongs to group IB phospholipase A(2) enzymes. Two short segments flanked by proline brackets are found in the sequence of MiPLA-1. These segments are on the surface of the molecule and hence may be involved in protein-protein recognition. PMID- 10462456 TI - Concepts and transformational knowledge. AB - The effect of exposure to principled change in concept formation was investigated in four experiments. In Experiment 1, participants were trained on either patterns that transformed systematically or control patterns that were distorted randomly. Training on transformational patterns produced concepts that were more resistant to false intrusions and decay. Experiment 2 separated the relative influences of transformational knowledge and pairwise similarity. Participants were able to identify the next pattern in a transformational sequence even though the foils were closer to the training patterns. Experiment 3 investigated whether participants use transformational information in a speeded categorization task. Participants were faster at classifying patterns that continued a transformational path than patterns that fell off the path, only if they had trained on the transformational patterns in a systematic order. Experiment 4 used multidimensional scaling to explore the psychological structure of transformational knowledge following training. Analyses revealed clear evidence of a transformational path with systematic training. Implications for theories of similarity and categorization are discussed. PMID- 10462458 TI - Hormone therapy: time for replacement? PMID- 10462457 TI - 2.5-month-old infants' reasoning about when objects should and should not be occluded. AB - The present research examined 2.5-month-old infants' reasoning about occlusion events. Three experiments investigated infants' ability to predict whether an object should remain continuously hidden or become temporarily visible when passing behind an occluder with an opening in its midsection. In Experiment 1, the infants were habituated to a short toy mouse that moved back and forth behind a screen. Next, the infants saw two test events that were identical to the habituation event except that a portion of the screen's midsection was removed to create a large window. In one event (high-window event), the window extended from the screen's upper edge; the mouse was shorter than the bottom of the window and thus did not become visible when passing behind the screen. In the other event (low-window event), the window extended from the screen's lower edge; although the mouse was shorter than the top of the window and hence should have become fully visible when passing behind the screen, it never appeared in the window. The infants tended to look equally at the high- and low-window events, suggesting that they were not surprised when the mouse failed to appear in the low window. However, positive results were obtained in Experiment 2 when the low-window event was modified: a portion of the screen above the window was removed so that the left and right sections of the screen were no longer connected (two-screens event). The infants looked reliably longer at the two-screens than at the high window event. Together, the results of Experiments 1 and 2 suggested that, at 2.5 months of age, infants possess only very limited expectations about when objects should and should not be occluded. Specifically, infants expect objects (1) to become visible when passing between occluders and (2) to remain hidden when passing behind occluders, irrespective of whether these have openings extending from their upper or lower edges. Experiment 3 provided support for this interpretation. The implications of these findings for models of the origins and development of infants' knowledge about occlusion events are discussed. PMID- 10462459 TI - ACE inhibitors or AT(1) receptor blockers in heart failure? PMID- 10462460 TI - Electrophysiological mechanisms involved in the increased mortality of patients with cardiac insufficiency. PMID- 10462461 TI - New interventions for obstructive hypertrophic cardiomyopathy: promise and prudence. PMID- 10462462 TI - The ratio of post-valvular aorta to valvular orifice cross-sectional areas: a new haemodynamic index of clinical importance in patients with aortic stenosis? PMID- 10462463 TI - Arteriogenesis, the good and bad of it. PMID- 10462464 TI - Methods for evaluating coronary microvasculature in humans. PMID- 10462465 TI - Decreased melatonin synthesis in patients with coronary artery disease. AB - AIMS: Decreased night-time plasma levels of melatonin were recently reported in patients with coronary artery disease, and it was postulated that melatonin production may be impaired, due to a lack of synthesizing enzymes. However, since artefacts possibly influencing the release pattern were not taken into account, this interpretation was strongly criticized. We therefore carefully investigated night-time melatonin production in patients with coronary artery disease using an appropriate experimental approach. Furthermore, we examined the effect of beta blockers, a frequently used drug in coronary artery disease therapy. METHODS AND RESULTS: Forty-eight male patients with angiographically documented severe coronary artery disease, 24 of them taking beta-blockers daily in therapeutic dosages, were included. Eighteen age-matched men, with no evidence of coronary sclerosis, served as controls. To determine melatonin production, 6 sulfatoxymelatonin (aMT6s) was measured radioimmunologically from overnight urine. Urinary aMT6s concentration was significantly decreased in patients, and beta-blocker treatment did not further suppress melatonin production. CONCLUSIONS: The data obtained using this investigative approach provide clearcut evidence that melatonin production in patients with coronary artery disease is decreased. Whether a decreased melatonin level may be a predisposing factor for coronary artery disease, or whether the occurrence of coronary artery disease decreases melatonin synthesis remains to be determined. PMID- 10462466 TI - Angiotensin I-converting enzyme gene polymorphism, coronary artery disease and myocardial infarction. An angiographically controlled study. AB - OBJECTIVES: We investigated the association between insertion/deletion polymorphism of the angiotensin I-converting enzyme (ACE) gene, the presence and extent of coronary artery disease, and myocardial infarction. BACKGROUND: The D allele of the ACE gene has been associated with coronary artery disease and myocardial infarction, but this association has been challenged in epidemiological studies. METHODS: Nine hundred and sixty-nine men and 341 women undergoing coronary angiography were studied. The ACE genotypes were assessed by polymerase chain reaction from genomic deoxyribonucleic acid, homozygosity for the D allele was controlled using an insertion-specific primer. Coronary artery disease was defined by angiographic criteria, the extent of coronary artery disease by the number of coronary arteries with >/=50% lumen narrowing. RESULTS: The ACE genotypes did not differ in terms of age, sex, body mass index, blood pressure, plasma lipids or lipoproteins. We found no association between the ACE genotypes and coronary artery disease (odds ratio, 95% confidence interval in DD genotypes for coronary artery disease in men 0.97, 0.70-1.36; in women 1.56, 0.95 2.57), extent of coronary artery disease (men 1.17, 0.85-1.61; women 1.24, 0.65 2.34), or myocardial infarction among the patients with coronary artery disease (men 1.07, 0.78-1.48; women 0.95, 0.50-1.76). The ACE genotype was not associated with coronary artery disease or myocardial infarction in hypertensives (n=771; odds ratio for coronary artery disease 0.93, 0.65-1.34; odds ratio for myocardial infarction 0.94, 0.66-1.33), or in patients GTP > guanosine 5'-[beta,gamma-imido]triphosphate > uridine 5'-[gamma-thio]triphosphate > UTP > CTP. Nucleoside 5'-diphosphates inhibited GTP hydrolysis in the order of potency GDP approximately guanosine 5'-[beta thio]thiodiphosphate > uridine 5'-[beta-thio]diphosphate >> UDP (no effect). UTP inhibited GTP hydrolysis competitively, indicative for nucleotide binding to the same site. Uracil nucleotides had a distinct activity profile with respect to disruption of the transitory complex between photoexcited rhodopsin and nucleotide-free transducin. We conclude that (i) uracil nucleotides bind to transducin-alpha with lower affinity than the corresponding guanine nucleotides, (ii) phosphorothioate modification of uracil nucleotides increases their affinity for transducin, and (iii) uracil nucleotides induce conformational changes in G proteins that are different from the conformational changes induced by guanine nucleotides. PMID- 10462478 TI - Activation of Rac1 increases c-Jun NH(2)-terminal kinase activity and DNA fragmentation in a calcium-dependent manner in rat myoblast cell line H9c2. AB - We examined the role of intracellular Ca(2+) in c-Jun NH(2)-terminal kinase (JNK) activation and DNA fragmentation in the rat myoblast cell line H9c2 using small GTP-binding protein Rac1. A constitutively active mutant of Rac1 (V12-Rac1) increased JNK-responsive gene expression 6-fold, although this increase was attenuated by the intracellular Ca(2+) chelator BAPTA-AM. V12-Rac1 also increased the number of DNA fragmentated cells. However, V12-Rac1-mediated JNK activation was not affected by BAPTA-AM as determined by direct measurement of active forms, and V12-Rac1 did not affect intracellular Ca(2+) concentration. These results suggest that Rac1 can activate JNK and induces cell injury, but [Ca(2+)](i) is necessary for V12-Rac1 to induce DNA fragmentation downstream of JNK activation. PMID- 10462479 TI - Nitric oxide inhibits ornithine decarboxylase by S-nitrosylation. AB - Ornithine decarboxylase (ODC) is the initial enzyme in the polyamine synthetic pathway, and polyamines are required for cell proliferation. We have shown previously that nitric oxide (NO) inhibits ODC activity in Caco-2 cells and in crude cell lysate preparations. In this study we examined the mechanism by which NO inhibits the activity of purified ODC. NO, in the form of S-nitrosocysteine (CysNO), S-nitrosoglutathione (GSNO), or 1, 1-diethyl-2-hydroxy-2-nitroso hydrazine (DEA/NO), inhibited enzyme activity in a concentration-dependent manner. CysNO (1 microM) inhibited ODC activity by approximately 90% and 3 microM GSNO by more than 70%. DEA/NO was less potent, inhibiting enzyme activity by 70% at a concentration of 30 microM. Inhibition of enzyme activity by CysNO, GSNO, or DEA/NO was reversible by addition of dithiothreitol or glutathione. Cuprous ion (Cu (I)) also reversed the inhibitory effect of these NO donor agents. The data presented here support the hypothesis that NO inhibits ODC activity via S nitrosylation of a critical cysteine residue(s) on ODC. PMID- 10462480 TI - Inorganic phosphate regulates CryIVA protoxin expression in Bacillus thuringiensis israelensis. AB - The role of nutritional factors during CryIVA protoxin expression in Bacillus thuringiensis israelensis (Bti) has been investigated. Inorganic phosphate (Pi) was found to stimulate 135 kD protoxin synthesis by Bti cells. There was a corresponding increase in the cryIVA specific mRNA in the presence of Pi. Inorganic phosphate inhibited HPr kinase but activated HPr phosphatase, the two enzymes responsible for regulating the concentration of phosphorylated HPr in the cell. Addition of protein phosphatase inhibitors NaF and calyculin A during resuspension resulted in the inhibition of toxin synthesis by Bti cells. Calyculin A inhibited HPr phosphatase activity in the in vitro assay also. The concentration of phosphorylated HPr was upregulated when the cells were resuspended in the presence of calyculin A, while the levels of the same were lowered in the presence of Pi, as determined by Western blotting the respective cells. The efficiency of sporulation of Bti was not affected when Pi was added alone or along with the phosphatase inhibitor calyculin A. PMID- 10462481 TI - A novel strategy for identifying differential gene expression: An improved method of differential display analysis. AB - We propose a novel alternative approach, an advanced method for recently developed strategies, for identifying differentially expressed genes. Firstly, double-stranded cDNAs were digested using Sau3AI and the 3'-end restriction fragments of the cDNA were ligated to a double-stranded adapter. Next, the restriction fragments were directly amplified using several combinations of adapter-specific primers and FITC-labeled oligo dT primers. The selected cDNA fragments were displayed on a polyacrylamide gel. Neither nested PCR nor purification of 3'-end fragments are necessary. We examined the validity of this approach by evaluating gene expression changes during granulocytic differentiation of HL-60 cells. This method can theoretically detect almost all gene expression changes more rapidly and through simpler manipulations than by any other approach. PMID- 10462482 TI - Cloning and characterization of psu1(+), a new essential fission yeast gene involved in cell wall synthesis. AB - We have isolated a new gene, psu1(+), from the fission yeast Schizosaccharomyces pombe. The predicted amino acid sequences shows that this protein has striking homology to the SUN family of the budding yeast, hence designated Psu1 (S. pombe homologue of the SUN family). Disruption of the psu1(+) gene revealed that it is essential for growth, and the null phenotype showed the swelling of cells followed by eventual lysis. We introduced psu1(+) gene in the disruptant strain and repressed it giving resistance to 1, 3-beta-glucanase digestion. Our results suggest that Psu1 plays an essential role in cell wall synthesis in S. pombe. PMID- 10462483 TI - Sesquiterpene lactones inhibit inducible nitric oxide synthase gene expression in cultured rat aortic smooth muscle cells. AB - Nitric oxide (NO) is an important regulator and effector molecule in various inflammatory disease states. High output of NO during inflammation is generated by the inducible isoform of nitric oxide synthase (iNOS). Sesquiterpene lactones are derived from Mexican-Indian medicinal plants and are known to have potent anti-inflammatory properties. The mechanisms by which sesquiterpene lactones exert their anti-inflammatory effects are not fully understood. In the current studies we determined if the sesquiterpene lactones, parthenolide and isohelenin, modulate iNOS gene expression in cultured rat aortic smooth muscle cells (RASMC) treated with lipopolysaccharide and interferon-gamma. Treatment with parthenolide or isohelenin inhibited NO production and iNOS mRNA expression in a concentration dependent manner. Transient transfection studies with an iNOS promoter-luciferase reporter plasmid demonstrated that parthenolide and isohelenin also inhibited activation of the iNOS promoter. Inhibition of iNOS promoter activation was associated with inhibition of both I-kappaBalpha degradation and nuclear translocation of NF-kappaB. Neither parthenolide nor isohelenin induced the heat shock response in RASMC. We conclude that sesquiterpene lactones inhibit iNOS gene expression by a mechanism involving stabilization of the I-kappaBalpha/NF kappaB complex. This effect is not related to induction of the heat shock response. The ability of sesquiterpene lactones to inhibit iNOS gene expression may account, in part, for their anti-inflammatory effects. PMID- 10462484 TI - Differences in expression of pro-caspases in small cell and non-small cell lung carcinoma. AB - Expression of several molecular determinants of apoptosis was analyzed in 10 untreated small cell (SCLC) and 6 untreated non-small cell (NSCLC) lung carcinoma cell lines. Although SCLC lines were more prone to spontaneous apoptosis compared with NSCLC lines, the former showed higher Bcl-2 expression and a higher Bcl 2/Bax ratio. In order to understand this apparent contradiction, the expression of pro-caspases as well as calpain was analyzed in these cell lines at the protein and mRNA levels. No differences in protein level of pro-caspases-2, -3, 7, and -9 and of calpain were detected between the SCLC and the NSCLC lines, but a striking difference in pro-caspase-8 expression was noted. All 6 NSCLC, but only 2 of the 10 SCLC lines, expressed pro-caspase-8 protein. Further experiments using the RNase protection assay indicated that the lack of pro-caspase-8 expression at the mRNA level was characteristic for SCLC. Using the same experimental approach, we found that SCLC cell lines in addition to pro-caspase-8 were deficient in mRNA expression of pro-caspases-1, -4, and -10, suggesting a different caspase-activating cascade in SCLC compared with NSCLC. This first systematic characterization of pro-caspase expression in lung cancer surprisingly showed that SCLC, which are more prone to undergo spontaneous apoptosis, are deficient in several pro-caspases and have a high Bcl-2/Bax ratio. Thus, the propensity of SCLC cells to undergo apoptosis cannot be explained only by the expression of factors involved in regulation or execution of apoptosis. PMID- 10462485 TI - Identification and characterization of human Fas associated factor 1, hFAF1. AB - We have identified and characterized a cDNA encoding human Fas associated factor 1 (hFAF1) cDNA and a shorter form of hFAF1 cDNA [hFAF1(s)] with a 456 bp internal in-frame deletion from a human HeLa cDNA library. The nucleotide sequences of hFAF1 and hFAF1(s) were identical except for the deletion. GST-hFAF1 fusion protein bound to the in vitro translation product of Fas. The N-terminal region (amino acid 1 approximately 201) including the upstream ubiquitin homology domain of hFAF1 could bind with the death domain of Fas unlike that of qFAF1 whose binding region with Fas could not be determined. However hFAF1 did not bind to the death domain of Fas mutant, lpr(cg). hFAF1 was expressed abundantly in testis, skeletal muscle, and heart as 2.8 kb mRNA. Polyclonal antibody against hFAF1 detected 74 kD protein, a deduced protein size from the ORF and 40 kD protein in some cell lines. PMID- 10462486 TI - Acquisition of ordered conformation by the N-terminal domain of the human small proline rich 2 protein. AB - The cornified cell envelope (CE) is a crucial structure for barrier function in terminally differentiated dead stratified squamous epithelia. It is assembled by transglutaminase enzymes (TGases) that cross-link several proteins such as loricrin and the small proline rich (SPR) proteins. Human SPR2 protein is cross linked with widely differing efficiencies by TGases 1, 2, and 3 using exclusively residues in the N- and C-terminal domains. In order to understand if the absence of the cross-linking catalyzed by TGases in the central domain is due to the conformation adopted, we have investigated the structural properties in solution of three peptides that correspond to the N-terminal domain, to three repeats of the central domain, and to the C-terminal domain. Together, the NMR and CD data strongly indicate the presence of a highly flexible non alpha-helix, non beta sheet structure in SPR2. Thus, SPR2 appears to function as a flexible cross bridging protein to provide tensile strength or rigidity to the CE of the stratified squamous epithelia in which it is expressed. PMID- 10462487 TI - Anomeric specificity and protein-substrate interactions support the 3D model for the hemagglutinin-neuraminidase from sendai virus. AB - The 3D structure of paramyxovirus hemagglutinin-neuraminidase has not yet been resolved; however, a theoretical model has been built by using influenza virus and bacterial neuraminidases as template [V. C. Epa (1997) Proteins Struct. Funct. Gen. 29, 264-281]. Two common features of the catalytic mechanism of the neuraminidases of known 3D structure are the anomeric specificity and the involvement of a tyrosine residue in the stabilization of the transition state. These key features have been investigated on the water-soluble ectodomain of the hemagglutinin-neuraminidase from Sendai virus (cHN). The anomeric specificity of the hydrolysis of the substrate by cHN has been investigated by NMR spectroscopy. The immediate product of the reaction was the alpha-anomer, meaning that cHN belongs between glycohydrolases retaining anomeric configuration like influenza virus neuraminidase. Measurements of the UV difference spectrum upon binding of the substrate analogue 2,3-dehydro 2-deossi N-acetyl neuraminic acid indicate the ionization of a tyrosine residue and decreased polarity in the environment of a tryptophan residue. Functional significance of the spectral data was derived from the known structure of influenza neuraminidase, where a tyrosinate ion is involved in the stabilization of the transition-state carbonium ion, and a tryptophan residue is involved in the binding of the acetyl moiety of the substrate. The data give experimental support to the 3D model of paramyxovirus neuraminidase. PMID- 10462488 TI - Fast laser-induced solute release from liposomes sensitized with photochromic lipid: effects of temperature, lipid host, and sensitizer concentration. AB - Liposomes of gel-phase phospholipid have been prepared containing a photochromic lipid sensitizer. A fast UV laser pulse isomerizes the sensitizer destabilizing the lipid bilayer structure and causing release of trapped solute. The kinetics of solute release have been investigated as a function of host lipid chain length, sensitizer concentration, and temperature, and the limits of liposome stability have been established. At low concentrations of sensitizer, pulsed laser irradiation induces some solute release when continuous UV illumination is ineffective. Although rates of solute release usually increase with temperature, at low sensitizer concentration in a rigid host, leakage at first increases but then decreases rapidly above a threshold temperature. The results presented are relevant to the design of photostimulated drug delivery systems and to potential applications of photosensitive liposomes as caging agents for biological effectors. PMID- 10462489 TI - Structural analysis of the titin gene in hypertrophic cardiomyopathy: identification of a novel disease gene. AB - Hypertrophic cardiomyopathy (HCM) is characterized by ventricular hypertrophy accompanied by myofibrillar disarrays. Molecular genetic analyses have revealed that mutations in 8 different genes cause HCM. Mutations in these disease genes, however, could be found in about half of HCM patients, suggesting that there are other unknown disease gene(s). Because the known disease genes encode sarcomeric proteins expressed in the cardiac muscle, we searched for a disease-associated mutation in the titin gene in 82 HCM patients who had no mutation in the known disease genes. A G to T transversion in codon 740, from CGC to CTC, replacing Arginine with Leucine was found in a patient. This mutation was not found in more than 500 normal chromosomes and increased the binding affinity of titin to alpha actitin in the yeast two-hybrid assay. These observations suggest that the titin mutation may cause HCM in this patient via altered affinity to alpha-actinin. PMID- 10462490 TI - Substance P regulates the function of rabbit cultured osteoclast; increase of intracellular free calcium concentration and enhancement of bone resorption. AB - The effects of substance P on cultured rabbit osteoclasts were investigated. The intracellular Ca(2+) concentration ([Ca(2+)](i)) which was monitored by the microfluorometric technique using fura-2, in osteoclasts elevated by the addition of substance P (0.3-5 microM). The EC(50) value of substance P was about 200 nM. This substance P-evoked [Ca(2+)](i) elevation was not observed in the Ca(2+)-free medium. Simultaneous application of spantide, a substance P receptor antagonist, blocked the Ca(2+) response. The addition of substance P (0.1-10 microM) to cultured osteoclasts enhanced their bone resorption activity which was evaluated by the pit formation assay. Maximum enhancement of the pit formation by substance P (5 microM) peaked at about 170% of the control level. The addition of substance P receptor antagonists also inhibited the enhancement of the bone resorption by substance P addition. Substance P possibly stimulates the bone remodeling by osteoclasts via the [Ca(2+)](i) elevation. PMID- 10462491 TI - The importance of L-phenylalanine transport and its autocrine turnover to L tyrosine for melanogenesis in human epidermal melanocytes. AB - A comparative study of (14)C-labeled l-phenylalanine and (3)H-labeled L-tyrosine uptake in cultures of human melanocytes (n = 8) and keratinocytes (n = 2) identified a significantly more rapid active transport for L-phenylalanine in melanocytes, whereas the slower uptake of L-tyrosine followed a concentration dependent gradient, confirming facilitated diffusion rather than active transport. In addition, a significantly more efficient autocrine turnover of L phenylalanine to L-tyrosine via intracellular phenylalanine hydroxylase was demonstrated in melanocytes. The incorporation of the newly synthesized radiolabeled L-tyrosine was directly followed into the end product melanin. The presence of L-phenylalanine in the culture medium produced 40% more melanin compared to an equivalent concentration of L-tyrosine alone. The transport of extracellular L-phenylalanine and its intracellular metabolism to L-tyrosine via intracellular phenylalanine hydroxylase are coupled to calcium uptake/efflux, whereas L-tyrosine uptake is calcium independent. Taken together, our results identified for the first time the importance of autocrine calcium-dependent active l-phenylalanine uptake/turnover in melanocytes as a major pathway for melanogenesis. PMID- 10462492 TI - Human T-cell lymphotrophic virus type-I tax gene induces secretion of human macrophage inflammatory protein-1alpha. AB - Human T-cell lymphotropic virus I (HTLV-I) encodes for a 40-kDa protein, Tax, which is important for the immortalization of T cells. Tax has been shown to transactivate several cellular genes. In this study, we show that MIP-1alpha is selectively expressed and secreted in the tax transfected Jurkat cell line upon mitogen stimulation. Expression of MIP-1alpha-R mRNA in these cells suggests an autocrine role for this chemokine in HTLV-I infected T-cells. Induced MIP-1alpha expression and secretion in PMA/PHA stimulated tax transfected cells correlate with the noninduction of MNP-1 transcription factor, which is intimately involved in downmodulating the MIP-1alpha gene. PMID- 10462493 TI - Hepatoma-derived growth factor-related protein (HRP)-1 gene in spermatogenesis in mice. AB - Hepatoma-derived growth factor (HDGF)-related protein (HRP)-1, a member of the HDGF gene family, showed testis-specific expression in mice. HRP-1 expression in spermatogenesis was analyzed in the testis of normal and azoospermic mice by Northern blot and immunohistochemistry. HRP-1 gene message was not expressed in the ovary and its product was detected only in the nuclei of germ cells, not in somatic cells. The HRP-1 gene is expressed through pachytene spermatocyte to round spermatid. HRP-1 gene expression was not detected in the testis of cryptorchid mice or in some strains of mutant mice. These findings suggest that the testis-specific HRP-1 gene may play an important role in the phase around meiotic cell division. PMID- 10462494 TI - Mouse phosphoinositide 3-kinase p110alpha gene: cloning, structural organization, and localization to chromosome 3 band B. AB - Phosphoinositide 3-Kinases (PI3-Kinases) are a family of dual specificity enzymes with a unique lipid kinase activity toward the D-3 position of the inositol ring of phosphoinositides and a less well characterized serine/threonine protein kinase activity. Class IA PI3-Kinases comprise a 110-120 kDa catalytic subunit (usually termed p110) and an 85 kDa or 50 to 55 kDa regulatory subunit (often called p85). cDNAs for three mammalian Class IA PI3-Kinase catalytic subunits designated p110alpha, p110beta, and p110delta have been cloned from several species. A YAC clone for the human p110alpha gene has also been cloned and mapped to chromosome 3q26.3. However, structural organization for any of the PI3-Kinase p110alpha genes has not been reported. Here, we report the cloning, structural organization, and chromosomal localization of the mouse PI3-Kinase p110alpha gene. The translated portion of the mouse p110alpha gene is encoded by 19 exons that span at least 24 kb. Dual color fluorescence in situ hybridization (FISH) was performed to determine the chromosomal localization of the mouse PI3-Kinase p110alpha gene. FISH results and DAPI banding demonstrated localization of the p110alpha gene to band B on mouse chromosome 3, a region syntenic with human chromosome 3q26.3. PMID- 10462495 TI - Hedgehog signaling molecules in bone marrow cells at the initial stage of fracture repair. AB - Ihh is a secreted protein expressed in chondrocytes in cartilaginous soft callus and thought to be involved in regulation of chondrogenic differentiation in fracture repair processes. However, gene expression and function of Ihh and its signaling molecules, Ptc and Smo, at the initial stage of fracture repair remain unknown. In the present study, we showed by RT-PCR of mouse rib fractures that the upregulation of Ihh mRNA occurred within hours after fracture, immediately followed by that of Ptc mRNA, and that both Ihh and Ptc mRNAs exhibited the time course similar to those of OP and OC mRNAs at the initial stage of fracture repair. The transcript level of Smo mRNA gradually increased within hours after fracture and was continuously maintained throughout the subsequent fracture repair processes. By in situ hybridization analysis, the transcripts of Ptc and Smo genes localized in bone marrow of unfractured ribs, and those of Ihh, Ptc, and Smo were expressed in the vicinity of the fracture site at 8 h after fracture. Furthermore, in adherent bone marrow cells in culture, mrIhh-N upregulated the gene expression of TGF-beta(1) as well as OPGL, a potent stimulator of osteoclastogenesis and osteoclast activity. These observations suggest that Ihh may play roles in the initial stage of fracture repair via TGF beta(1) and OPGL. PMID- 10462497 TI - Substitution of the 2'-hydroxyl group at position 2.1 by an amino group interferes with Mg(2+) binding and efficient cleavage by hammerhead ribozyme. AB - Recently we have demonstrated that hammerhead ribozymes can be fully substituted with 2'-amino pyrimidines without detriment to the catalytic activity, provided that positions 2.2 and/or 2.1 are not modified. We now report on the potential molecular mechanisms by which 2'-amino groups at these positions inhibit the ribozyme cleavage activity. In the presence of Mg(2+), the 2'-amino modification at positions 2.2 and/or 2.1 had no significant effect on substrate binding. Detailed analysis of the ribozyme initial cleavage rates in the presence of various Mg(2+) concentrations indicated that Mg(2+) binding is inhibited by the 2'-amino group at position 2.1. Furthermore, preannealed substrate molecules to the modified ribozyme are not effectively cleaved upon Mg(2+) addition, indicating an alteration of the ribozyme cleavage step. Surprisingly, the cleavage activity of the modified ribozymes was substantially increased when Mg(2+) ions were replaced by the thiophilic Mn(2+) ions, whereas only a moderate cleavage enhancement occurred with its unmodified version. Taken together, our findings indicate that changes in the sugar at position 2.1 alter Mg(2+) promoting ribozyme cleavage. PMID- 10462496 TI - Characterization of the promoter region of the human melanocortin-1 receptor (MC1R) gene. AB - We sequenced 3201 bp upstream from the ATG translation start codon of the human melanocortin-1 receptor (MC1R). A number of transcriptional initiation sites were detected over a region of approximately 600 base pairs upstream of the receptor coding region. These consist of GC-rich regions, each including SP-1 consensus binding motifs. Neither a TATA nor a CAAT box was found in this region. The 5' flanking region also contains the consensus regulatory elements for AP-1, AP-2, and several E-boxes. Gel shift assays targeting the three GC boxes confirmed binding of SP-1. A promoter assay revealed that the minimal region exhibiting promoter activity was located between nucleotides -517 and -282 in human melanoma SK-Mel-2 cells. Further deletion from -517 to -447, which removed an SP-1 site, completely abolished luciferase activity. In conclusion, the MC1R promoter shares the characteristics of many other GPCR promoters. These characteristics include GC-rich sequence, lack of a TATA box, and binding of SP-1. PMID- 10462498 TI - Differential utilization of cyclic ADP-ribose pathway by chemokines to induce the mobilization of intracellular calcium in NK cells. AB - We show here that cyclic adenosine diphosphate-ribose (cADPR) may be a second messenger for chemokines. Extracts collected from NK cells stimulated with IL-8 for 2 min were incubated with beta-NAD for an additional 2 min (designated as IL 8 extracts). This mixture elevated the mobilization of (Ca(2+))(i) in alpha-toxin permeabilized NK cells. This activity was inhibited upon prior incubation of these cells with ruthenium red but not with heparin. Purified cADPR and not Ins 1,4,5 P(3) desensitized NK cells to the calcium mobilization effect of IL-8 extracts. Further analysis showed that ruthenium red and heparin differentially inhibit RANTES-, SDF-1alpha-, or MDC-induced calcium mobilization in IL-2 activated NK cells. Also, introduction of anti-ryanodine receptor antibody inside streptolysin O-permeabilized NK cells resulted in complete inhibition of MDC, and only partial inhibition of RANTES and SDF-1alpha-induced calcium fluxes in NK cells. Collectively, these results suggest that chemokines may utilize the cADPR/ryanodine receptor pathway as well as the Ins 1,4,5 P(3)/Ins 1,4,5 P(3) receptor signaling pathway to induce the accumulation of calcium in NK cells. PMID- 10462499 TI - Influence of N-glycan chain length on chaperone association and intracellular transport of major histocompatibility complex class I proteins. AB - Recent studies demonstrate that processing of N-linked glycans plays an important role in the quality control of major histocompatibility complex (MHC) class I transport from the endoplasmic reticulum (ER) to the Golgi complex and beyond. Here, we investigated the importance of oligosaccharide chain length on the association of MHC class I proteins with molecular chaperones and their intracellular transport from the ER to the Golgi. These data show that calnexin interaction with class I proteins having truncated N-glycans was reduced compared to normal class I molecules, whereas the assembly of class I with calreticulin and TAP was unperturbed by N-glycan chain length. Additionally, these results demonstrate that class I proteins containing truncated N-glycans showed decreased detachment from calreticulin and TAP relative to class I proteins bearing typical oligosaccharides. Taken together, these studies show that N-glycan chain length is an important determinant for the quality control of newly synthesized MHC class I proteins in the ER. PMID- 10462500 TI - Regulation of oscillations in filamentous actin content in polymorphonuclear leukocytes stimulated with leukotriene B(4) and platelet-activating factor. AB - Stimulation of neutrophils with LTB(4) or PAF results in the production of a rapidly oscillating actin polymerization/depolymerization response. Treatment of neutrophils with inhibitors of PKC prior to stimulation with ligand resulted in a masking of the F-actin oscillations. Because myosin has been shown to be a substrate for neutrophil PKC, this protein was investigated as a potential downstream mediator of F-actin oscillations. Stimulation of neutrophils with LTB(4) resulted in myosin light chain being serine phosphorylated in a PKC dependent manner. This phosphorylation was shown to occur in a manner that is kinetically distinct from the myosin phosphorylation induced by FMLP, a potent activator of actin polymerization that alone does not induce F-actin oscillations. Additionally, disruption of intracellular actin-myosin interactions resulted in inhibition of LTB(4)- as well as PAF-induced F-actin oscillations. These data suggest that PKC and downstream phosphorylation of myosin as well as actin-myosin interaction may play roles in mediating the production of neutrophil F-actin oscillations. PMID- 10462501 TI - The molecular basis for the B(A) allele: an amino acid alteration in the human histoblood group B alpha-(1,3)-galactosyltransferase increases its intrinsic alpha-(1,3)-N-acetylgalactosaminyltransferase activity. AB - The formation of the subgroup B(A) phenotype is thought to be due to an overlapping specificity of the human blood group A and B transferases. A new molecular basis for the B(A) allele, resulting from the C(700) to G substitution which predicts the alteration of Pro(234) to Ala, just ahead of the second of the four amino acid residues which differentiates the specificities of the A and B transferases, is reported here. Compared to normal group B sera, a relatively lower B-transferase activity was demonstrated in the B(A) serum, which correlated well with the observation of a smaller amount of B antigen on the B(A) red cells. Also a much higher A-transferase activity was demonstrated in the B(A) serum in contrast to the minute amount of A-transferase activity found in normal group B sera. The formation of the B(A) phenotype in this report is most likely due to the shifting of the specificity of the B transferase rather than an enhanced B transferase activity which was previously presumed to be responsible for the formation of this phenotype. The Pro(234) to Ala alteration is suggested to be responsible for the shifting of the specificity with a subsequent increase in A- but a decrease in B-transferase activity. This new B(A) allele shows that not only the four critical residues but also the neighboring areas may influence the specificity of the A and B transferases. PMID- 10462502 TI - Transcriptional suppression of rat angiotensin AT1a receptor gene expression by interferon-gamma in vascular smooth muscle cells. AB - Angiotensin (Ang) II stimulates proliferation of vascular smooth muscle cells (VSMC) via its specific receptor AT1 subtype, possibly leading to atherosclerosis in hypertension. On the other hand, a cytokine interferon (IFN)-gamma has been shown to have an anti-atherosclerotic effect. In the present study, we examined a possible role of IFN-gamma in AT1 receptor gene regulation in VSMC. A firefly luciferase expression vector driven by the rat AT1a receptor gene promoter ( approximately 3.2 kb) was transfected into the cultured rat VSMC, and luciferase expression was determined to estimate the transcription function of the AT1a receptor gene promoter. RT-PCR was also carried out to determine mRNA expression of AT1a receptor in VSMC. IFN-gamma treatment decreased AT1a receptor mRNA expression as well as luciferase expression in a dose-dependent manner. The analysis with deletion DNA fragments showed that the IFN-responsive element was located between -987 and -331 positions, where multiple GAS (gamma interferon activated site)-like elements were identified. The expression suppression was reversed by either a MAPKK inhibitor PD98059 or a Jak-2 inhibitor AG-490. These results suggest that IFN-gamma can inhibit AT1 receptor expression at gene transcription level, and that the transcription suppression is dependent on MAP kinase and Jak-2. Inhibition of AT1a receptor expression may possibly be implicated in the anti-atherosclerotic action of IFN-gamma in VSMC. PMID- 10462503 TI - Relationship between mRNA levels quantified by reverse transcription-competitive PCR and metabolic activity of CYP3A4 and CYP2E1 in human liver. AB - Reverse transcription-competitive polymerase chain reaction is a powerful and sensitive tool for quantifying the absolute amount of mRNA. Using this method with beta-actin as the standard, we measured the mRNA level of CYP3A and CYP2E1 isoforms in human livers. We also determined the metabolic activities for both CYP isoforms. The absolute amounts of CYP3A4 and CYP2E1 mRNA in 15 liver tissues ranged from 15 to 5127 and 15163 to 69289 copies/10(4) copies of beta-actin (341- & 3.6-fold), respectively. The testosterone 6beta-hydroxylation for CYP3A4 and chlorzoxazone 6-hydroxylation activity for CYP2E1 ranged from 30 to 505 pmol/mg/min (16-fold) and from 0.59 to 2.73 nmol/mg/ml (3.6-fold), respectively. The correlation between the mRNA level and activity of CYP3A4 was significant (r = 0.94), while there was no significant correlation for CYP2E1 (r = 0.04). In conclusion, we observed a significant correlation between enzyme activity and mRNA expression for CYP3A4 but not for CYP2E1. This fact indicates that CYP2E1, in addition to being less variable between individuals than CYP3A4, differs in its regulation mechanism. PMID- 10462504 TI - Functional heterogeneity of C-terminal peroxisome targeting signal 1 in PEX5 defective patients. AB - To investigate mechanisms related to functions of the peroxisome targeting signal (PTS) 1 receptor, Pex5p, we analyzed peroxisome matrix protein import in fibroblasts from three patients with peroxisome biogenesis disorders, all with different mutations in the PEX5 gene. The patients 2-01 (Zellweger syndrome) and 2-05 (neonatal adrenoleukodystrophy) have the reported mutations, R390X and N489K, and patient 2-03 (infantile Refsum disease) has a newly identified mutation, S563W. Fibroblasts from 2-03 (S563W) were detected in both PTS1 and PTS2 imports despite the PEX5 defect, findings in contrast with fibroblasts from 2-05 (N489K) severely defective in PTS1 import and those from 2-01 (R390X) severely defective in both PTS1 and PTS2. The PTS1 receptor in 2-03 is functional for only the C-terminal -SKL sequence (acyl-CoA oxidase) and had little or no function for C-terminal -AKL (D-bifunctional protein and sterol carrier protein 2) and -KANL (catalase) sequences, respectively. After transfection of these mutated PEX5 cDNA into the PEX5-defective CHO mutant, transformants of ZP102 revealed that each mutation was responsible for each dysfunction of the PTS1 import. It seems apparent that -AKL and -KANL are poorer variants of PTS1 and are likely to be more susceptible to effects of mutation of its receptor, Pex5p. PMID- 10462505 TI - Characterization of cis-elements required for the transcriptional activation of the rae28/mph1 gene in F9 cells. AB - The rae28/mph1 gene is the mouse homologue of the Drosophila polyhomeotic gene, which plays a crucial role in the maintenance of the transcriptional repression state of Hox genes. Expression of the rae28/mph1 gene is induced during retinoic acid (RA)-mediated differentiation of embryonal carcinoma F9 cells. By transient transfection experiments, we identified a pair of inverted differentiation response sequences (DRS(s)) in the 5' flanking region. Each of the DRS(s) contained the consensus sequence [5'-CCTCCCCXCXGCCCCCTCCXCXC-3'], which is also conserved in the human counterpart of the rae28/mph1 gene. Electrophoretic mobility shift assay and DNase I foot printing with nuclear extracts derived from F9 cells demonstrated the presence of novel DNA-binding factors which specifically interact with DRS(s). Nucleotide substitutions in the 3' DRS abrogated the factor binding and the transcriptional activation, suggesting that DRS(s) and DRS-binding factors play an important role in the transcriptional regulation of the rae28/mph1 gene. PMID- 10462506 TI - Involvement of c-Jun NH(2)-terminal kinase pathway in differential regulation of heat shock proteins by anticancer drugs. AB - In the present study, we examined the modulation of heat shock factor 1 (HSF1) activity and expression of heat shock proteins (HSPs) after exposure to anticancer drugs. Anticancer drugs induced HSF1 DNA-binding activity, and this was followed by an increase of mitochondrial HSP75 and HSP60 levels and concurrent decrease of cytoplasmic HSP70 levels. Unlike heat shock-induced full phosphorylation, HSF1 was partially phosphorylated after exposure to vincristine, and this result was tightly correlated with the kinetics of JNK/SAPK activation, and up-regulation of mitochondrial HSP75 level and concurrent down-regulation of HSP70. Furthermore, the dominant-negative mutant of SEK1 blocked the phosphorylation of HSF1 and up-regulation of mitochondrial HSP75 in response to vincristine or vinblastine. These data suggest that anticancer drugs regulate the HSF1 transcriptional activity differently from heat shock, and JNK/SAPK pathway appears to be involved in anticancer drug-induced HSF1 phosphorylation and consequently differential regulation of mitochondrial HSP75 and HSP60 and cytoplasmic HSP70. PMID- 10462507 TI - Sequences at the 3' side of the c-fos SRE mediate gene expression via an Sob1 dependent, TCF-independent pathway. AB - We previously described a 110-kDa tyrosine phosphoprotein, Sob 1, that regulates formation of the DNA binding complex Band A at the c-fos serum response element (SRE) during T cell activation. Using competition and mutant oligonucleotide analysis, we have determined that both the core CArG box of the c-fos SRE and the 3' sequences flanking the CArG box are necessary for stable Band A complex formation. Moreover, using transient transfection and reporter assays, we show that mutations affecting Band A complex formation in vitro also impaired serum induction of c-fos gene expression in vivo. Since mutation at this site has no effect on SRF binding, our results suggest that in combination with SRE/SRF, Sob 1-regulated factor(s) bind at the 3' side of SRE to form Band A, and this confers maximal serum induction of c-fos gene expression via the SRE. PMID- 10462510 TI - Immunological relatedness of chicken and human riboflavin carrier protein. AB - Polyclonal antibodies (pAbs) to riboflavin carrier protein (RCP) purified from human amniotic fluid were raised in two rabbits. These pAbs apart from recognizing the parent antigen, reacted significantly with chicken RCP as well. When all the 212 octapeptides overlapping by one amino acid residue and covering the entire primary amino acid sequence of chicken RCP immobilized on polyethylene rods were probed with rabbit anti-human RCP antibodies, it was found that these antibodies recognize five epitopic sequences (residues 37-42, 73-76, 133-140, 174 176 and 200-207) in chicken RCP primary structure common to both the rabbits. In addition, the antisera from the two individual animals also recognized additional epitopes characteristic to that particular animal. The reactivities of these antigenic determinants have been confirmed with polyclonal anti-human RCP sera by using synthetic peptides harbouring these chicken RCP sequences in inhibition ELISA. This high level of immunological similarities among linear epitopes observed may reflect the degree of conservation of local conformations among linear antigenic determinants shared by the two evolutionarily distant homologous proteins. PMID- 10462509 TI - Rb independent inhibition of cell growth by p15(INK4B). AB - The INK4 cyclin dependent kinase inhibitors (CDKI), such as p15(INK4B) and p16(INK4A), block cell cycle progression from G to S phase. This is mediated by inhibition of phosphorylation of proteins, including the retinoblastoma susceptibility protein (Rb), by cyclin dependent kinases. Ectopic over-expression of the p16(INK4A) CDKI can inhibit growth of cell lines depending on Rb status. Cell lines lacking Rb, with few exceptions, are resistant to growth inhibition by p16(INK4A). The effects of ectopic over-expression of p15(INK4B) in cell lines with and without wild type Rb were examined by measuring cell recovery. Proliferation was inhibited in cells lacking Rb as well as in cells with wild type Rb expression. Experiments analyzing the effectiveness of chimeric p15(INK4B)/p16(INK4A) proteins indicated that the Rb independent growth inhibition required N-terminal residues of p15(INK4B). Linker insertion mutation of p15(INK4B) showed that the inhibition was dependent on intact ankyrin structures. Double staining flow cytometry found that the growth inhibition correlated with a decrease in cells in G2/M phases of the cell cycle. These findings are consistent with Rb independent inhibition of the progression from G1 to S caused by overexpression of p15(INK4B). PMID- 10462508 TI - Replication of hepatitis B virus which carries foreign DNA in vitro. AB - Targeting a specific DNA sequence to the desired tissues is an important step in gene therapy. The hepatitis B virus (HBV) is the only DNA virus that has hepatocyte specificity. We attempted to construct an HBV-based vector for targeting the liver. We observed the replication and secretion of virus particles in an HBV construct that lacks X gene and carries an extra 63 bp DNA fragment in vitro. Replication was observed in the cell line HuH-7 but not HepG2. From this construct, we designed an HBV-based vector that could carry foreign DNA. HBV based vectors provide for the possibilities of generating therapeutic agents for individual patients. Our host vector system may be used to clear out the HBV from the HBV carrier or chronic hepatitis B patients by introducing a genetically engineered HBV into these patients. PMID- 10462511 TI - Nitric oxide synthase imunolabeling in the molluscan CNS and peripheral tissues. AB - NOS immunoreactivity was assayed in CNS and peripheral tissues of the sea slugs Pleurobranchaea californica, Tritonia diomedea and Aplysia californica using different antisera against mammalian nitric oxide synthase in Western blots. Polyclonal anti-nNOS labeled at 250, 185, 170, 155, 100, 75, and 65 kD in extracts of Pleurobranchaea CNS, salivary gland and esophagus but not of gills or muscle. The labeling pattern for Tritonia in bands at 250, 200, 120/110, 100, 69, 65, and 60 kD differed somewhat. Anti-nNOS labeling in Aplysia was markedly different, with bands labeled only at 69 and 60 kD in CNS extracts, and at 200, 190, 69 and 60 kD in salivary and esophagus extracts. The wide variation in NOS immunoreactivity is consistent with species differences in tissue localization and biochemical properties of molluscan NOS isoforms. PMID- 10462512 TI - trans-sialidase of Trypanosoma cruzi: location of galactose-binding site(s). AB - Trypanosoma cruzi expresses a trans-sialidase on its surface, which catalyzes the transfer of sialic acid from mammalian host glycans to its own surface glycoproteins. It has been proposed that the enzyme consists of three domains prior to a long C-terminal repeating sequence that is not required for enzyme activity. The first of these domains shares significant sequence identity with bacterial sialidases which catalyse the hydrolysis of sialic acid. Here we report the sequence of the N-terminal domains of the TS19y trans-sialidase gene, which was expressed in bacteria with the same specific activity as natural enzyme of T. cruzi. Various deletion mutants of TS19y, without the C-terminal tandem repeat, have been cloned and expressed and their trans-sialidase and sialidase activities measured. These experiments show that all three N-terminal domains are required for full trans-sialidase activity, though only the first is necessary for sialidase activity. Some transferase activity is observed, however, even with the shortest construct comprising the first N-terminal domain. Deletion mutants to probe the role of the N-terminal residues of the first domain suggest that the first 33 residues are also required for trans-sialidase activity, but not for sialidase activity. Molecular modelling of the first N-terminal domain of TS19y based on our structures of bacterial sialidases and site-directed mutations suggests the location of a galactose-binding site within this domain. PMID- 10462513 TI - A new member of the leucyl aminopeptidase family purified and identified from a marine unicellular algae. AB - Leucyl aminopeptidase (LAP; EC 3.4.11.1) activity was purified from crude extracts of the marine unicellular algae Gonyaulax polyedra by a combination of hydrophobic interaction with phenyl sepharose, DEAE-cellulose, and mono-Q HR5/5 ion-exchange chromatography. The undenaturated protein has a molecular mass of about 110 kD and based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the enzyme appears to be composed of two possibly identical subunits of 55 kD. The identity of the protein was confirmed by a cross-reaction of the purified protein with an antibody raised against a commercial LAP. Biochemical characterization showed that the Gonyaulax enzyme was similar to most of the previously described LAPs. Gonyaulax LAP is a metallo-enzyme since EDTA and 1,10-phenathroline significantly inhibited activity. Addition of the metal ions Zn(2+), Cu(2+) inhibited 80% of LAP activity, suggesting they are not the natural cofactors of the enzyme. Other metals, such as Ca(2+), Co(2+), Mn(2+), or Mg(2+) (concentrations up to 4 mM), caused no alteration in the total activity of Gonyaulax LAP. PMID- 10462514 TI - Kinetic analysis of substrate inhibition in nitric oxide reductase of Paracoccus denitrificans. AB - The current kinetic model for the nitric oxide reductase reaction (Girsch, P., and de Vries, S. (1997) Biochim. Biophys. Acta 1318, 202-216) does not involve the concentration of an electron donor. Here we introduce this variable and show, both theoretically and experimentally, its role in determining the extent of substrate inhibition by the excess of nitric oxide. NO is found to inhibit competitively with the electron donor, possibly by binding to the oxidized form of the enzyme. The observed partial character of the inhibition is tentatively explained by a slow reduction of the non-productive NO complex. PMID- 10462515 TI - Inhibitory effects of vitamin A and vitamin K on rat cytochrome P4501A1-dependent monooxygenase activity. AB - The inhibitory effects of vitamins A and K toward P4501A1-dependent 7 ethoxycoumarin O-deethylation were examined in the reconstituted system containing the microsomal fraction prepared from the recombinant Saccharomyces cerevisiae cells producing rat P4501A1 and yeast NADPH-P450 reductase. On vitamins A, all-trans-retinol, all-trans-retinal, all-trans-retinoic acid and retinol-palmitate showed competitive inhibition with K(i) values of 0.068, 0.079, 2.6 and 2.0 microM, respectively. Judging from the K(i) values, the inhibitory effects of those vitamins A appear to have physiological significance on the basis of their contents in liver, lung and kidney. On vitamins K, vitamin K(1) showed competitive inhibition with K(i) value of 24 microM, while vitamin K(2) showed noncompetitive inhibition with K(i) value of 60 microM. Judging from these K(i) values together with the contents of these vitamins K in liver, the inhibitory effects of the vitamins K are not as significant as those of vitamins A. These results suggest that the ingestion of enough amounts of vitamins A from foods might lead to the inhibition of the activity of P4501A1 which is known to be induced by smoking, drugs such as omeprazole and lansoprazole, and environmental pollutants like dioxins. PMID- 10462516 TI - Leishmania species: models of intracellular parasitism. AB - Leishmania species are obligate intracellular parasites of cells of the macrophage-dendritic cell lineage. Indeed, the ability to survive and multiply within macrophages is a feature of a surprising number of infectious agents of major importance to public health, including Mycobacterium tuberculosis, Mycobacterium leprae, Listeria monocytogenes, Salmonella typhimurium, Toxoplasma gondii and Trypanosoma cruzi. The relationship between such organisms and their host cells is particularly intriguing because, not only are macrophages capable of potent microbicidal activity, but in their antigen-presenting capacity they can orchestrate the developing immune response. Thus, to initiate a successful infection parasites must gain entry into macrophages, and also withstand or circumvent their killing and degradative functions. However, to sustain a chronic infection, parasites must also subvert macrophage-accessory-cell activities and ablate the development of protective immunity. The leishmanias produce a wide spectrum of disease in mice, and as such they have provided excellent models for studying problems associated with intracellular parasitism. In recent years, largely using these organisms, we have made enormous progress in elucidating the mechanisms by which successful intracellular infection occurs. Furthermore, characterization of immunological pathways that are responsible for resistance or susceptibility to Leishmania has given rise to the Th1/Th2 paradigm of cellular/humoral dominance of the immune response. PMID- 10462517 TI - PECAM-1 (CD31) functions as a reservoir for and a modulator of tyrosine phosphorylated beta-catenin. AB - Catenins function as regulators of cellular signaling events in addition to their previously documented roles in adherens junction formation and function. Evidence to date suggests that beta and gamma catenins can act as signaling molecules, bind transcriptional factors and translocate to the nucleus. Beta- and gamma catenin are also major substrates for protein tyrosine kinases, and tyrosine phosphorylation of junctional proteins is correlated with decreased adhesiveness. One way in which catenin functions are modulated is by dynamic incorporation into junctional complexes which controls, in part, the cytoplasmic levels of catenins. Here we show that: (1) vascular endothelial growth factor (VEGF) induces beta catenin tyrosine phosphorylation in a time-, and dose-dependent manner and that VEGF receptors co-localize to areas of endothelial cell-cell contact in vitro and in vivo. (2) Platelet-endothelial cell adhesion molecule (PECAM)-1 can function as a reservoir for, and modulator of, tyrosine phosphorylated beta-catenin. (3) PECAM-1 can prevent beta-catenin nuclear translocation in transfected SW480 colon carcinoma cells. We suggest that PECAM-1 may play a role in modulating beta catenin tyrosine phosphorylation levels, localization and signaling and by doing so, functions as an important modulator of the endothelium. PMID- 10462518 TI - NrCAM, cerebellar granule cell receptor for the neuronal adhesion molecule F3, displays an actin-dependent mobility in growth cones. AB - The neuronal adhesion glycoprotein F3 is a multifunctional molecule of the immunoglobulin superfamily that displays heterophilic binding activities. In the present study, NrCAM was identified as the functional receptor mediating the inhibitory effect of F3 on axonal elongation from cerebellar granule cells. F3Fc conjugated microspheres binding to neuronal growth cones resulted from heterophilic interaction with NrCAM but not with L1. Time-lapse video-microscopy indicated that F3Fc beads bind at the leading edge and move retrogradely to reach the base of the growth cone within a lapse of 30-60 seconds. Such velocity (5.7 microm/minute) is consistent with a coupling between F3 receptors and the retrograde flow of actin filaments. When actin filaments were disrupted by cytochalasin B, the F3Fc beads remained immobile at the leading edge. The retrograde mobility appeared to be dependent on NrCAM clustering since it was induced upon binding with cross-linked but not dimeric F3Fc chimera. These data indicate that F3 may control growth cone motility by modulating the linkage of its receptor, NrCAM, to the cytoskeleton. They provide further insights into the mechanisms by which GPI-anchored adhesion molecules may exert an inhibitory effect on axonal elongation. PMID- 10462519 TI - A mouse model of intestinal stem cell function and regeneration. AB - We present here an in vivo mouse model for intestinal stem cell function and differentiation that uses postnatal intestinal epithelial cell aggregates to generate a differentiated murine small intestinal mucosa with full crypt-villus architecture. The process of neomucosal formation is highly similar to that of intestinal regeneration. Both in vivo grafting and primary culture of these cells reveal two different epithelial cell populations, which display properties consistent with intestinal epithelial transit amplifying and stem cell populations. Using this model system with a mixture of wild-type and transgene marked cells, we have shown that neomucosae originally develop from single aggregates, but that over time the mucosae fuse to form chimaeric mucosae. Despite fusion, the chimaeric mucosae maintain crypt clonality and villus polyclonality, demonstrating that clonal segregation persists during intestinal epithelial regeneration. PMID- 10462520 TI - Basonuclin is associated with the ribosomal RNA genes on human keratinocyte mitotic chromosomes. AB - Basonuclin is a zinc finger protein mainly expressed in keratinocytes of the basal layer of epidermis and the outer root sheath of hair follicles. It is also found in abundance in the germ cells of testis and ovary. In cultured keratinocytes, basonuclin is associated with chromatin in all phases of the cell cycle, including mitosis. By immunocytochemical methods, we demonstrate here that in mitosis basonuclin is associated with the short arms of the acrocentric chromosomes and with other loci on many metaphase chromosomes of human keratinocytes. Using the evolutionarily highly conserved N-terminal pair of zinc fingers in an electrophoresis mobility shift assay, we demonstrate that the DNA target sequences of basonuclin on the acrocentric chromosomes are likely to be within the promoter region of the 45S rRNA gene transcription unit. DNase I footprinting shows that basonuclin zinc fingers interact with the upstream control element of this promoter, which is necessary for the high level of transcription of the rRNA genes. This result suggests that basonuclin may be a tissue-specific transcription factor for the ribosomal RNA genes. PMID- 10462521 TI - PBC68: a nuclear pore complex protein that associates reversibly with the mitotic spindle. AB - Using autoimmune antibodies from a patient with primary biliary cirrhosis we have identified a 68 kDa nuclear envelope protein, termed PBC68. This protein is co precipitated with a 98 kDa and a 250 kDa polypeptide and is distinct from the nuclear lamins. Immunostaining of digitonin-permeabilized cells indicates that PBC68 is restricted to the inner (nucleoplasmic) face of the nuclear envelope, while indirect immunofluorescence and immunoelectron microscopy show that PBC68 is located on fibrillar structures emanating from the nuclear pore complex. The autoantigen is modified at early prophase and disassembles at prometaphase concurrently with the breakdown of the nuclear envelope. The disassembled material, instead of diffusing throughout the cytoplasm as other nucleoporins, is targeted to the mitotic spindle and remains stably bound to it until anaphase. At telophase PBC68 is released from the mitotic apparatus and reassembles late, after incorporation of LAP2B and B-type lamins, onto the reforming nuclear envelope. The partitioning of PBC68 in dividing cells supports the notion that subsets of nuclear envelope proteins are actively sorted during mitosis by transiently anchoring to spindle microtubules. Furthermore, the data suggest that specific constituents of pore complex are released in a stepwise fashion from their anchorage sites before becoming available for nuclear reassembly. PMID- 10462522 TI - Characterization of tGLP-1, a Golgi and lysosome-associated, transmembrane glycoprotein of African trypanosomes. AB - Purification of endosomal/lysosomal vesicles of Trypanosoma brucei brucei bloodstream forms and generation of monoclonal antibodies led to the isolation of antibodies directed against an 85 kDa, Golgi and endocytic traffic-associated protein termed tGLP-1, Trypanosoma Golgi/lysosome protein-1. Preliminary immunoelectron microscopical analysis revealed that the protein is present in, but not restricted to, the limiting membrane of multivesicular lysosomes and is more abundant in bloodstream forms compared to the procyclic stage. The corresponding gene was cloned and is present as a single copy. Blast searches did not reveal any homologies to other proteins and genes published. The nucleotide sequence of the gene (1848 base pairs) predicted a type 1 membrane topology with an N-terminal signal sequence (20 aa), a luminal domain with 2 N-glycosylation sites (524 aa), a transmembrane domain (23 aa), and a long cytosolic tail domain (49 aa). Polyclonal antibodies raised against the cytosolic tail confirmed the localization of the gene product to multivesicular lysosomes but revealed that the majority of the protein was in the Golgi apparatus. Colabelling with an antibody against p67, a lysosomal glycoprotein of trypanosomes, revealed extensive overlap between the proteins with opposing relative abundance. Expression of the tGLP-1 open reading frame in Leishmania resulted in Golgi localization, and in Toxoplasma, in localization to both the Golgi and endoplasmic reticulum. These data indicate conservation in the functionality of the Golgi-targeting sequence of tGLP-1. PMID- 10462523 TI - Volume of liquid below the epithelium of an F9 cell as a signal for differentiation into visceral endoderm. AB - When retinoic acid-primed F9 cells are allowed to aggregate, they form embryoid bodies with an outer layer of (&agr;)-fetoprotein-producing visceral endoderm cells and an internal cavity. I show that maturation of the visceral endoderm is dependent on the size of F9 aggregates. Size fractionation of aggregates of retinoic acid-primed F9 cells on Percoll density gradients revealed that only aggregates with diameters larger than 180 microm developed into embryoid bodies with an endoderm layer secreting (&agr;)-fetoprotein. Size dependent alpha fetoprotein-secretion was also observed when retinoic acid-primed F9 cells were cultured on porous microcarrier beads larger than 185 microm. Retinoic acid primed F9 cells on flat microporous membranes did not differentiate and secrete alpha-fetoprotein unless exposed to a limited volume of medium at their basolateral surface. This suggested that maturation of the visceral endoderm is signaled by the volume of liquid phase below the epithelium. I postulate that the epithelial layer of an F9 aggregate encloses liquid and forms a barrier to diffusion of some critical factor(s). The concentration of such a factor may reach a threshold due to enlargement of the liquid phase during growth of the F9 aggregate and thereby signal maturation of the outer layer of cells into visceral endoderm. PMID- 10462524 TI - Ezrin regulates cell-cell and cell-matrix adhesion, a possible role with E cadherin/beta-catenin. AB - Ezrin, radixin, moesin and merlin form a subfamily of conserved proteins in the band 4.1 superfamily. The function of these proteins is to link the plasma membrane to the actin cytoskeleton. Merlin is defective or absent in schwannomas and meningiomas and has been suggested to function as a tumour suppressor. In this study, we have examined the role of ezrin as a potential regulator of the adhesive and invasive behaviour of tumour cells. We have shown that following inhibition of ezrin expression in colo-rectal cancer cells using antisense oligonucleotides, these cells displayed a reduced cell-cell adhesiveness together with a gain in their motile and invasive behaviour. These cells also displayed increased spreading over matrix-coated surfaces. Immunofluorescence studies revealed that antisense-treated cells also displayed an increased staining of paxillin in areas representing focal adhesions. Furthermore, coprecipitation studies revealed an association of ezrin with E-cadherin and beta-catenin. Induction of the phosphorylation of ezrin by orthovanadate and hepatocyte growth factor/scatter factor resulted in changes similar to those seen with antisense treatment, together with a marked decrease in the association of ezrin with both beta-catenin and E-cadherin. It is concluded that ezrin regulates cell-cell and cell-matrix adhesion, by interacting with cell adhesion molecules E-cadherin and beta-catenin, and may thus play an important role in the control of adhesion and invasiveness of cancer cells. PMID- 10462525 TI - A novel protein targeting domain directs proteins to the anterior cytoplasmic face of the flagellar pocket in African trypanosomes. AB - The flagellar pocket of African trypanosomes is a critical sorting station for protein and membrane trafficking, and is considered to be an Achilles' heel of this deadly pathogen. Although several proteins, including receptors for host derived growth factors, are targeted specifically to the flagellar pocket, the signals responsible for this restricted subcellular localization are entirely unknown. Using T lymphocyte triggering factor-green fluorescent protein (TLTF(1) GFP) fusion proteins, we demonstrate that an internal 144 amino acid domain of TLTF from Trypanosoma brucei is sufficient for directing GFP to the cytoplasmic side of the anterior flagellar pocket. Immuno-gold electron microscopy reveals that the TLTF-GFP fusion protein is located in an electron dense structure that immediately abuts the anterior flagellar pocket membrane. The amino acid sequence of the TLTF targeting domain does not resemble previously characterized protein trafficking signals, and random mutagenesis reveals that flagellar pocket targeting is conferred by a structural motif, rather than a short, contiguous array of amino acids. The aberrant sorting of two mutant proteins into the flagellum, and the targeting of a related human protein to the plus end of the trypanosome's cytoskeletal microtubules, lead us to suggest that flagellar pocket targeting involves interactions with the trypanosome cytoskeleton. The finding that TLTF-GFP is restricted to the anterior, cytoplasmic face of the flagellar pocket membrane, suggests that there is structural heterogeneity in the membrane of this organelle. PMID- 10462526 TI - The S. pombe zfs1 gene is required to prevent septation if mitotic progression is inhibited. AB - Schizosaccharomyces pombe cdc16p is required to limit the cell to forming a single division septum per cell cycle; the heat-sensitive loss-of-function mutant cdc16-116 completes mitosis, and then undergoes multiple rounds of septum formation without cell cleavage. cdc16p is a homologue of Saccharomyces cerevisiae BUB2p, and has also been implicated in the spindle assembly checkpoint function in S. pombe. To identify other proteins involved in regulating septum formation, we have screened for multicopy suppressors of the cdc16-116 mutation. In this paper, we describe one of these suppressors, zfs1. The null allele (zfs1 D1) is viable. However, at low temperatures it divides at a reduced size, while at higher temperatures, it partially suppresses heat sensitive mutants in genes signalling the onset of septum formation. Zfs1-D1 cells show an increased rate of chromosome loss during exponential growth. Moreover, if assembly of the spindle is prevented, zfs1-D1 cells do not arrest normally, but the activity of cdc2p kinase decays, and cells form a division septum without completing a normal mitosis. We conclude that zfs1 function is required to prevent septum formation and exit from mitosis if the mitotic spindle is not assembled. The suppression of cdc16-116 by zfs1 is independent of dma1 function and the spindle assembly checkpoint genes mad2 and mph1. The genetic interactions of zfs1 with genes regulating septum formation suggest that it may be a modulator of the signal transduction network controlling the onset of septum formation and exit from mitosis. PMID- 10462527 TI - Two di-leucine-based motifs account for the different subcellular localizations of the human endothelin-converting enzyme (ECE-1) isoforms. AB - Endothelin-converting enzyme (ECE-1) is a type II integral membrane protein which plays a key role in the biosynthetic pathway of the vasoconstricting endothelins. Three ECE-1 isoforms, differing by their N-terminal cytoplasmic tails, are generated from a single gene. When expressed in CHO cells, they display comparable enzymatic activity but whereas ECE-1a is strongly expressed at the cell surface, ECE-1b is exclusively intracellular and ECE-1c presents an intermediate distribution. In the present study these different localizations were further described at the ultrastructural level, by electron microscope immunocytochemistry. To characterize the motifs responsible for the intracellular localization of ECE-1b we constructed chimeric proteins and point mutants. Two di leucine-based motifs, contained in the N-terminal part of ECE-1b, were thus identified. One of these motifs (LV), displayed by both ECE-1b and ECE-1c, accounts for the reduced surface expression of ECE-1c as compared to ECE-1a. Mutation of both motifs (LL and LV) induces a very strong appearance of ECE-1b at the cell surface indicating that their presence in the N-terminal extremity of ECE-1b is critical for its exclusively intracellular localization. PMID- 10462528 TI - Functional domains of human growth hormone necessary for the adipogenic activity of hGH/hPL chimeric molecules. AB - Genetic analysis through construction of chimeric genes and their transfection in mammalian cells could provide a better understanding of biological functions of native or modified proteins, and would allow the design of new gene constructs encoding peptides that mimic or block ligand interaction with target tissues. To identify the hGH domains responsible for induction of adipose differentiation we constructed hGH/hPL chimeric molecules using homologous DNA mutagenesis, since hGH, but not human placental lactogen (hPL), promotes adipose differentiation in mouse 3T3-F442A cells. We assayed their adipogenic activity in an autocrine/paracrine biological model consisting of transiently transfected 3T3 F442A cells with the chimeric constructs. Plasmid DNAs carrying these constructs were transfected into growing 3T3-F442A cells, and cultures were further maintained for 7 days to differentiate into adipocytes. Secretion of transfected hGH/hPL chimeric proteins into the medium was in the range of 5-25 ng/ml. Adipogenic activity was a property only of those chimeric proteins that contained hGH exon III together with either hGH exon II or hGH IV. Our results also suggest that hGH binding site-2 is composed of two structural subdomains: subsite 2A encoded by exon II of hGH and subsite-2B encoded by exon IV. We also suggest that full adipogenic activity requires the presence of binding site-1 and any of the subsites of binding site-2. This simple autocrine/paracrine biological model of gene transfection allows the analysis of specific biological activity of products encoded by modified genes. PMID- 10462529 TI - Translational control of the cdc25 cell cycle phosphatase: a molecular mechanism coupling mitosis to cell growth. AB - The eukaryotic translation initiation factor 4A (eIF4A) is an RNA helicase required for translation initiation of eukaryotic mRNAs. By engineering fission yeast mutants with diminished eIF4A activity, we have found that translation of cdc25 mRNAs (a dosage-dependent activator of mitosis in all eukaryotic cells) is particularly sensitive to limitations of protein synthesis mediated by limited eIF4A activity. Genetic and biochemical analysis indicated that a rate-limited translation initiation of cdc25 mRNAs, exerted throughout its unusual 5' untranslated leader, acts as a molecular sensor to ensure that a minimum cell mass (protein synthesis) is attained before mitosis occurs. The Cdc13 cyclin B is also among the limited pool of proteins whose translation is sensitive to reduced translation initiation activity. Interestingly, the 5' leader sequences of cdc25 and cdc13 mRNAs have conserved features which are unusual in other yeast mRNAs, suggesting that common mechanisms operate in the expression of these two key mitotic activators at the translational level. PMID- 10462530 TI - Requirement of nuclear factor kappaB for the constitutive expression of nitric oxide synthase-2 and cyclooxygenase-2 in rat trophoblasts. AB - Recently isolated trophoblasts express nitric oxide synthase 2 (NOS-2) and cyclooxygenase 2 (COX-2), decreasing the levels of the corresponding mRNAs when the cells were maintained in culture. The sustained expression of COX-2 and NOS-2 in trophoblasts was dependent on the activation of nuclear factor kappaB (NF kappaB) since proteasome inhibitors and antioxidants that abrogated NF-kappaB activity suppressed the induction of both genes. The time-dependent fall of the mRNA levels of NOS-2 and COX-2 paralleled the inhibition of NF-kappaB, determined by electrophoretic mobility shift assays, and the increase of the IkappaBalpha and IkappaBbeta inhibitory proteins. Isolated trophoblasts synthesized reactive oxygen intermediates (ROI), a process impaired after culturing the cells, and that might be involved in the NF-kappaB activation process. Moreover, treatment of recently isolated cells with ROI scavengers suppressed the expression of COX-2 and NOS-2. Challenge of trophoblasts with interleukin-1beta up-regulated the expression of both proteins, an effect that was potentiated by lipopolysaccharide. These results indicate that the physiological expression of NOS-2 and COX-2 in trophoblasts involves a sustained activation of NF-kappaB which inhibition abrogates the inducibility of both genes. PMID- 10462531 TI - Mechanical loading regulates tenascin-C expression in the osteotendinous junction. AB - Elastic extracellular matrix protein tenascin-C (TN) has very restricted expression in normal tissues, but is expressed in large quantities during embryogenesis and hyperplastic processes. To examine the importance of mechanical stress on the regulation of TN expression in vivo, the effects of various mechanical loading states (immobilization and three forms of subsequent remobilization) on the expression of TN were studied immunohistochemically at the bone-tendon attachment of the rat quadriceps muscle. This osteotendinous junction (OTJ) was selected as study site, since it receives its mechanical stimuli only from muscle contracting activity, which is easy to block by cast immobilization. TN was expressed abundantly in the normal OTJ. Following the removal of the mechanical stress from the junction by cast-immobilization of three weeks, the immunoreactivity of TN was almost completely absent. Normal mechanical stress in the form of free remobilization of eight weeks (free cage activity) resulted in a slight increase in TN expression, but could not restore the expression of TN to the level of the healthy contralateral leg. After the application of the increased mechanical stress (intensified remobilization of the eight weeks by low or high-intensity treadmill running), the distribution and immunoreactivity of TN reached the level of the healthy contralateral limb in the low-intensity running group or even exceeded that in the high-intensity running group. High TN expression was seen around the chondrocytes and fibroblasts of the OTJ as well as around the collagen fibers of the tendon belly. We conclusively show that mechanical strain regulates the expression of TN in vivo, and propose that mechanical stress is a major regulator of TN expression in fibroblasts and chondrocytes. This may be an important aspect of the regulation of TN expression during embryogenesis, tendon degeneration, wound healing, bone formation, and in the other normal or regenerative morphogenetic processes TN is postulated to take part in. PMID- 10462532 TI - Dietary glycine and renal denervation prevents cyclosporin A-induced hydroxyl radical production in rat kidney. AB - Cyclosporin A (CsA) nephrotoxicity is associated with renal hypoxia and increases in free radicals in the urine. This study was designed to elucidate the mechanism of radical production caused by CsA. Pretreatment of rats with CsA (25 mg/kg, i.g.) for 5 days decreased glomerular filtration rates by 65%, an effect largely prevented by both dietary glycine (5%) or renal denervation. CsA dissolved in olive oil produced a 6-line alpha-(4-pyridyl 1-oxide)-N-tert-butylnitrone (4 POBN)/free radical signal in the urine, which partitioned predominantly into the aqueous phase after chloroform extraction (i.e., it is water soluble). Dimethyl sulfoxide (DMSO) is attacked by the hydroxyl radical to produce a methyl radical; administration of CsA with [(12)C]DMSO produced two radical species in urine, one with hyperfine coupling constants similar to the 4-POBN/methyl radical adduct found in aqueous solution. CsA given with [(13)C]DMSO produced a 12-line spectrum, confirming the formation of hydroxyl radicals. The methyl radical produced by the hydroxyl radical represented 62% of radicals detected in urine but only 15% in bile. Therefore, hydroxyl radicals are produced largely in the kidney. Free radicals in urine were increased about 5-fold by CsA, an effect completely blocked by the inhibitory neurotransmitter, glycine, or by renal denervation. CsA infusion for 30 min increased efferent renal nerve activity 2 fold, and dietary glycine (5%) totally blocked this phenomenon. Taken together, these data are consistent with the hypothesis that CsA increases hydroxyl radical formation by increasing renal nerve activity resulting in vasoconstriction and hypoxia-reoxygenation. Glycine blunts the effect of CsA on the renal nerve, which explains, in part, prevention of nephrotoxicity. PMID- 10462533 TI - A novel domain of the inhibitory glycine receptor determining antagonist efficacies: further evidence for partial agonism resulting from self-inhibition. AB - Different amino side chains in the N-terminal extracellular region of the inhibitory glycine receptor (GlyR) have been shown to be crucial for ligand recognition. Here we describe a novel domain of the GlyRalpha1 subunit that constitutes an important determinant of antagonist activity. The antagonists strychnine, nipecotic acid, and isobutyric acid displayed reduced potencies at recombinant GlyRs formed from alpha1 subunits, in which lysine 104, phenylalanine 108, or threonine 112 were replaced by alanine. Agonist affinities, in contrast, were slightly increased at these mutant receptors. Taurine and beta aminoisobutyric acid, which are partial agonists at the wild-type GlyR, behaved as full agonists at the mutant GlyRs and failed to inhibit glycine-induced currents. This is consistent with apolar residues at positions 104, 108, and 112 of the alpha1 subunit reducing the antagonistic, but not the agonistic, binding of beta-amino acids. Our data support a model in which the partial agonism of beta-amino acids results from their self-inhibitory activity. PMID- 10462534 TI - Developmental action of estrogen receptor-alpha feminizes the growth hormone Stat5b pathway and expression of Cyp2a4 and Cyp2d9 genes in mouse liver. AB - We have studied the roles of estrogen receptor-alpha (ERalpha) and the Stat5b form of STAT (signal transducers and activators of transcription) in sex-specific expression of Cyp2a4 (steroid 15alpha-hydroxylase) and Cyp2d9 (steroid 16alpha hydroxylase) genes using ERalpha-deficient mice. ERalpha deficiency resulted in the repression of the female-specific Cyp2a4 and expression of the male-specific Cyp2d9 genes, respectively in females. In ERalpha-deficient males, the Cyp2d9 gene continued to be expressed. Nuclear localization of Stat5b occurs in both sexes of ERalpha-deficient mice, although it is normally observed in only wild type males. Nuclear localization of Stat5b correlates with the repression of Cyp2a4 and expression of Cyp2d9, respectively. Because Stat5b was not detectable in liver nuclear extracts prepared from hypophysectomized ERalpha-deficient females, the regulation by ERalpha appeared to be mediated through a pituitary hormone (i.e., growth hormone). Thus, ERalpha appears to play a key role in the mechanism that inhibits nuclear localization of Stat5b in female mice, leading to feminization of a ERalpha-GH-Stat5b pathway and Cyp expression. Defaulting to this ERalpha-dependent mechanism results in localization of Stat5b to nuclei, which masculinizes the expression of Cyp genes in male mice. PMID- 10462535 TI - DNA protein cross-links produced by NSC 652287, a novel thiophene derivative active against human renal cancer cells. AB - 2, 5-bis(5-Hydroxymethyl-2-thienyl)furan (NSC 652287), is a representative of a series of thiophene derivatives that exhibit potent and selective antitumor activity against several tumor cell lines in the National Cancer Institute Anticancer Drug Screen. NSC 652287 has noticeable activity for the renal cell lines and produces cures in certain corresponding xenografts. The cellular mechanisms of action of NSC 652287 were therefore investigated in this study in greater detail. The most sensitive renal carcinoma cell line, A498, exhibited cell cycle arrest in G(0)-G(1) and G(2)-M at 10 nM NSC 652287, with increased p53 and p21(WAF1) protein. At higher concentrations, NSC 652287 still induced p53 elevation but with p21(WAF1) reduction and massive apoptosis. These results collectively suggested that NSC 652287 induced DNA damage. Using alkaline elution techniques, we found that NSC 652287 induced both DNA-protein and DNA-DNA cross links with no detectable DNA single-strand breaks. These DNA-protein cross-links (DPC) persisted for at least 12 h after drug removal and their frequency was correlated with cytotoxicity in the renal cell lines studied. The most sensitive cells (A498) produced the highest DPC followed by the cell line with intermediate sensitivity (TK-10). DPC were minimal in the two resistant cell lines, ACHN and UO-31. Nonetheless, a similar degree of DPC occurred at doses imparting equitoxic effects. These results indicate that DNA is a primary target for the novel and potent anticancer thiophene derivative, NSC 652287. NSC 652287 did not cross-link purified DNA or mammalian topoisomerase I suggesting the importance of active metabolite(s) for the cross-linking activity. PMID- 10462536 TI - Spontaneous beta(2)-adrenergic signaling fails to modulate L-type Ca(2+) current in mouse ventricular myocytes. AB - A receptor can be activated either by specific ligand-directed changes in conformation or by intrinsic, spontaneous conformational change. In the beta(2) adrenergic receptor (AR) overexpression transgenic (TG4) murine heart, spontaneously activated beta(2)AR (beta(2)-R*) in the absence of ligands has been evidenced by elevated basal adenylyl cyclase activity and cardiac function. In the present study, we determined whether the signaling mediated by beta(2)-R* differs from that of a ligand-elicited beta(2)AR activation (beta(2)-LR*). In ventricular myocytes from TG4 mice, the properties of L-type Ca(2+) current (I(Ca)), a major effector of beta(2)-LR* signaling, was unaltered, despite a 2.5 fold increase in the basal cAMP level and a 1.9-fold increase in baseline contraction amplitude as compared with that of wild-type (WT) cells. Although the contractile response to beta(2)-R* in TG4 cells was abolished by a beta(2)AR inverse agonist, ICI118,551 (5 x 10(-7) M), or an inhibitory cAMP analog, Rp-CPT cAMPS (10(-4) M), no change was detected in the simultaneously recorded I(Ca). These results suggest that the increase in basal cAMP due to beta(2)-R*, while increasing contraction amplitude, does not affect I(Ca) characteristics. In contrast, the beta(2)AR agonist, zinterol elicited a substantial augmentation of I(Ca) in both TG4 and WT cells (pertussis toxin-treated), indicating that L-type Ca(2+) channel in these cells can respond to ligand-directed signaling. Furthermore, forskolin, an adenylyl cyclase activator, elicited similar dose dependent increase in I(Ca) amplitude in WT and TG4 cells, suggesting that the sensitivity of L-type Ca(2+) channel to cAMP-dependent modulation remains intact in TG4 cells. Thus, we conclude that beta(2)-R* bypasses I(Ca) to modulate contraction, and that beta(2)-LR* and beta(2)-R* exhibit different intracellular signaling and target protein specificity. PMID- 10462537 TI - Mechanism-based chemopreventive strategies against etoposide-induced acute myeloid leukemia: free radical/antioxidant approach. AB - Etoposide (VP-16) is extensively used to treat cancer, yet its efficacy is calamitously associated with an increased risk of secondary acute myelogenous leukemia. The mechanisms for the extremely high susceptibility of myeloid stem cells to the leukemogenic effects of etoposide have not been elucidated. We propose a mechanism to account for the etoposide-induced secondary acute myelogenous leukemia and nutritional strategies to prevent this complication of etoposide therapy. We hypothesize that etoposide phenoxyl radicals (etoposide O(.)) formed from etoposide by myeloperoxidase are responsible for its genotoxic effects in bone marrow progenitor cells, which contain constitutively high myeloperoxidase activity. Here, we used purified human myeloperoxidase, as well as human leukemia HL60 cells with high myeloperoxidase activity and provide evidence of the following. 1) Etoposide undergoes one-electron oxidation to etoposide-O(.) catalyzed by both purified myeloperoxidase and myeloperoxidase activity in HL60 cells; formation of etoposide-O(.)radicals is completely blocked by myeloperoxidase inhibitors, cyanide and azide. 2) Intracellular reductants, GSH and protein sulfhydryls (but not phospholipids), are involved in myeloperoxidase-catalyzed etoposide redox-cycling that oxidizes endogenous thiols; pretreatment of HL60 cells with a maleimide thiol reagent, ThioGlo1, prevents redox-cycling of etoposide-O(.) radicals and permits their direct electron paramagnetic resonance detection in cell homogenates. VP-16 redox cycling by purified myeloperoxidase (in the presence of GSH) or by myeloperoxidase activity in HL60 cells is accompanied by generation of thiyl radicals, GS(.), determined by HPLC assay of 5, 5-dimethyl-1-pyrroline glytathionyl N-oxide glytathionyl nitrone adducts. 3) Ascorbate directly reduces etoposide-O(.), thus competitively inhibiting etoposide-O(.)-induced thiol oxidation. Ascorbate also diminishes etoposide-induced topo II-DNA complex formation in myeloperoxidase-rich HL60 cells (but not in HL60 cells with myeloperoxidase activity depleted by pretreatment with succinyl acetone). 4) A vitamin E homolog, 2,2,5,7, 8-pentamethyl-6-hydroxychromane, a hindered phenolic compound whose phenoxyl radicals do not oxidize endogenous thiols, effectively competes with etoposide as a substrate for myeloperoxidase, thus preventing etoposide-O(.)-induced redox-cycling. We conclude that nutritional antioxidant strategies can be targeted at minimizing etoposide conversion to etoposide-O(.), thus minimizing the genotoxic effects of the radicals in bone marrow myelogenous progenitor cells, i.e., chemoprevention of etoposide-induced acute myelogenous leukemia. PMID- 10462538 TI - Suppression of flavin-containing monooxygenase by overproduced nitric oxide in rat liver. AB - Effects of excessive nitric oxide (NO) produced in vivo by an i.p. injection of bacterial lipopolysaccharide (LPS) on hepatic microsomal drug oxidation catalyzed by flavin-containing monooxygenase (FMO) were determined. At 6 and 24 h after the LPS injection, liver microsomes were isolated and FMO activities were determined by using FMO substrates like thiobenzamide, trimethylamine, N,N-dimethylaniline, and imipramine. Liver microsomal FMO activities of LPS-treated rats were decreased significantly for all these substrates. Microsomal content of FMO1 (the major form in rat liver) in LPS-treated rats as determined by immunoblotting, was severely decreased as well. In support of this, hepatic content of FMO1 mRNA was decreased by 43.6 to 67.3%. However, the hepatic content of inducible NO synthase (iNOS) mRNA was increased by 2.6- to 5.4-fold and the plasma nitrite/nitrate concentration was increased by about 30-fold in the LPS-treated rats. When this overproduction of NO in the LPS-treated rats was inhibited in vivo by a single or repeat doses of either a general NOS inhibitor N(G)-nitro-L-arginine or a specific iNOS inhibitor aminoguanidine, the FMO1 mRNA levels were not severely depressed (70-85% of the control level). Attendant with the reduction of plasma nitrite/nitrate concentration by single and repeated doses of NOS inhibitors, activity and content of FMO1 in liver microsomes isolated from these NOS inhibitor cotreated rats were restored partially (in single-dose inhibitors) or completely (in repeat doses). In contrast to these NO-mediated in vivo suppressive effects on the mRNA and enzyme contents of FMO1 as well as the FMO activity, the NO generated in vitro from sodium nitroprusside did not inhibit the FMO activities present in microsomes of rat and rabbit liver as well as those present in rabbit kidney and lung. Combined, the excessive NO produced in vivo (caused by the LPS-dependent induction of iNOS) suppresses the FMO1 mRNA and enzyme contents as well as the FMO activities without any direct in vitro effect on the activities of premade FMO enzyme. These findings suggest that NO is an important mediator involved in the suppression of FMO1 activity in vivo. Thus, together with the previously reported suppression on the cytochrome P-450 activities, the overproduced NO in the liver caused by induction of iNOS under conditions of endotoxemia or sepsis suppresses FMO and appears to be responsible for the decreased drug oxidation function observed generally under conditions of systemic bacterial or viral infections. PMID- 10462539 TI - Ras mutation, irrespective of cell type and p53 status, determines a cell's destiny to undergo apoptosis by okadaic acid, an inhibitor of protein phosphatase 1 and 2A. AB - Okadaic acid (OA), a toxin from the black sponge Halicondria okadai, is a specific inhibitor of serine/threonine protein phosphatases 1 (PP1) and 2A (PP2A). OA is a tumor promoter but also induces apoptosis in some tumor cell lines. In this study, we determined whether ras mutation and/or p53 status are characteristics associated with the cell's sensitivity to the induction of apoptosis by OA. Several cell lines that differed in ras and p53 mutations were treated with OA (10-100 nM). At 24 to 48 h after treatment, the percentage of cells undergoing apoptosis was quantitated. The cell lines with mutations in either H-ras (human bladder carcinoma cell line T24 and mouse keratinocyte cell line 308), or K-ras (human colon carcinoma cell lines DLD-1 and HCT116; human prostate cancer cell lines LNCaP and PC-3; human lung cancer cell lines Calu-6 and SKLU-1; and human pancreatic cancer cell line MIAPaCa2) were more sensitive to OA-induced apoptosis (3- to 10-fold) than the cell lines that lacked the ras mutation (mouse epidermal cell lines C50 and JB6; murine fibroblast cell line NIH3T3; human colon cancer cell line HT29; human kidney epithelial cell line Hs715.K; and human pancreatic cancer cell line Bx-PC3). Similarly, using isogenic cell lines we found that overexpression of mutated H-ras in NIH3T3 and in SV40 immortalized human uroepithelial cells (SVHUC) enhanced their sensitivity to undergo apoptosis in response to OA treatment. The T24, DLD-1, SKLU-1, Calu-6, and MIAPaCa2 cell lines express mutated p53. The SVHUC as well as their ras transfected counterparts have inactive p53 due to complex formation between large "T" antigen and p53. Taken together, these results imply that OA-induced apoptosis may involve a p53-independent pathway. The transfectants (NIH3T3-ras and SVHUC-ras), which express mutated H-ras, have up-regulated PP2A activity. OA treatment inhibited in vivo the levels of PP1 and PP2A activity, and induced apoptosis in SVHUC-ras and other cell lines. We conclude that OA-induced cell death pathway in ras-activated cell lines may involve a cross talk between PP1 and PP2A and ras signaling pathways. In light of the present results, the current theory that OA promotes mouse skin tumor formation by selective expansion of initiated cells that harbor ras mutations needs reevaluation. PMID- 10462540 TI - Activation of the mouse TATA-less and human TATA-containing UDP glucuronosyltransferase 1A1 promoters by hepatocyte nuclear factor 1. AB - UDP-glucuronosyltransferase (UGT) 1A1 (UGT1A1) catalyzes the glucuronidation of bilirubin in liver. Among all UGT isoforms identified to date, it is the only relevant bilirubin-glucuronidating enzyme in human. Because glucuronoconjugation is the major route of bilirubin elimination, any genetic alteration that affects bilirubin glucuronosyltransferase activity may result in a more or less severe hyperbilirubinemia. In this study, we report the cloning and characterization of the transcriptional regulation of the mouse UGT1A1 gene. Primary-structure analysis of the mouse Thymidine Adevice promoter revealed marked differences with its human homolog. First, the mouse promoter lacks the highly polymorphic thymidine/adenine repeat occurring in the human promoter, which has been associated with some forms of hyperbilirubinemia. Second, an L1 transposon element, which is absent in the human promoter, is found 480 bp upstream of the transcription start site in mouse. Using the electromobility shift and DNase I footprinting experiments, we have identified a hepatocyte nuclear factor 1 binding site in the mouse UGT1A1 promoter that confers responsiveness to both factors HNF1alpha and HNF1beta in HEK293 cells. Furthermore, we show that this element, which is conserved in the human promoter, also confers strong HNF1 responsiveness to the human UGT1A1 gene. Together, these results provide evidence for a major regulatory function of this liver-enriched transcription factor in UGT1A1 activity in both rodents and human. PMID- 10462541 TI - Post-translational regulation of AP-1 transcription factor DNA-binding activity in the rat conceptus. AB - Activator protein-1 (AP-1) transcription factor DNA binding is induced during transient oxidative stress in the midorganogenesis rat conceptus in culture. L-2 Oxothiazolidine-4-carboxylate (OTC), a cysteine prodrug, prevented oxidative stress and the induction of AP-1 binding activity in the embryo but not in the yolk sac. Because AP-1 activity may be a significant determinant of developmental outcome after insult, we investigated the regulation of AP-1 activity in the conceptus. Supershift assays indicated that basal AP-1 binding in the embryo was due primarily to JunD, whereas in the yolk sac c-Jun and JunD were important. Under oxidative stress, c-Fos and c-Jun contributed to the AP-1 binding in the embryo; in the yolk sac, a c-Fos-shifted complex emerged. OTC protection from oxidative stress did not change the AP-1 composition, suggesting that increased AP-1 activity was due to post-translational modifications. Changes in AP-1 activity in embryos under oxidative stress or with OTC protection were not the result of alterations in the net phosphorylation state of Fos or Jun proteins or of changes in activities of the extracellular signal-regulated kinases 1 and 2 or stress-activated protein kinases. However, immunodepletion of redox factor 1 (Ref 1), a nuclear factor that promotes AP-1 binding, eliminated AP-1 activity from embryonic nuclear extracts under both basal and oxidative stress conditions. Therefore, Ref-1 plays a critical role in regulating AP-1 activity in the conceptus; it is plausible that Ref-1-mediated modulation of the AP-1 stress response is a determinant of embryonic fate. PMID- 10462542 TI - Importance of the extracellular domain for prostaglandin EP(2) receptor function. AB - The ligand binding pocket of biogenic amine G protein-coupled receptors is embedded in the membrane-spanning regions of these receptors, whereas the extracellular domains of the peptidergic receptors play a key role in the structure and function of this class of receptors. To examine the role of the extracellular sequences in prostaglandin receptor-ligand interaction, chimeras were constructed with the two G(s)-coupled E-prostanoid (EP) receptors, replacing each of the extracellular sequences of the human EP(2) receptor with the corresponding human EP(4) receptor residues. Replacement of the third extracellular loop (ECIII) yielded a receptor that binds [(3)H]prostaglandin E(2) (PGE(2); K(d) = 6.3 nM) with similar affinity as the EP(2) wild-type receptor (K(d) = 12.9 nM). Similarly, replacement of the nonconserved carboxyl-terminal portion of ECII resulted in a receptor that maintains [(3)H]PGE(2) binding (K(d) = 8.8 nM). In contrast, replacement of the amino terminus, ECI, the entire ECII region, or the residues within the highly conserved motif of the amino-terminal half of ECII yielded chimeras that displayed neither detectable [(3)H]PGE(2) binding nor receptor-evoked cAMP generation. Immunoprecipitation demonstrated that each chimera is expressed at levels near that of wild-type receptors; however, enzyme-linked immunosorbent assay revealed that inactive chimeras have reduced cell surface expression. Similarly, chimeras that exchange the multiple extracellular loop sequences N/ECI, ECII/ECIII, or all four sequences lacked detectable binding and signal transduction, and although expressed, were not detected on the cell surface. These data suggest that the extracellular sequences of the EP(2) receptor are critical determinants of receptor structure and/or function, unlike other G protein-coupled receptors that bind small molecules. PMID- 10462543 TI - Pharmacological and molecular characterization of 5-hydroxytryptamine(7) receptors in the rat adrenal gland. AB - Serotonin (5-hydroxytryptamine; 5-HT) is a potent stimulator of aldosterone secretion in the rat adrenal gland but the type of receptor involved in the mechanism of action of 5-HT remains unknown. The aim of the present study was to determine the pharmacological profile and to clone the receptor responsible for the corticotropic effect of 5-HT in rat glomerulosa cells. A series of 10 serotonergic receptor agonists and 12 receptor antagonists was used to characterize the receptor mediating the effect of 5-HT on aldosterone secretion from perifused rat adrenocortical slices. Correlation analysis between the potencies of the different compounds in our model and those previously reported for various recombinant 5-HT receptors showed that the rat adrenal 5-HT receptor exhibits the same pharmacological profile as the 5-HT(7) receptor transiently expressed in COS-7 cells (r = 0.82 for agonists, p <.05; r = 0.83 for antagonists, p <.01). Polymerase chain reaction with specific primers revealed the expression of 5-HT(7) receptor mRNA in the rat adrenal gland. Cloning of the polymerase chain reaction product confirmed that the amplified DNA corresponded to the 5-HT(7) receptor cDNA sequence. Western blot analysis showed the presence of a protein with an apparent molecular mass of 66 kDa in the adrenal cortex but not in the medulla. Taken together, these data demonstrate that the rat adrenal glomerulosa expresses functional 5-HT(7) receptors. Rat glomerulosa cells will thus provide a robust and sensitive bioassay for future studies on native 5-HT(7) receptors. PMID- 10462544 TI - Phosphorylation of deoxycytidine analog monophosphates by UMP-CMP kinase: molecular characterization of the human enzyme. AB - Phosphorylation of deoxycytidine analogs by cellular enzymes is a prerequisite for the activity of these compounds. We have investigated the kinetic parameters for the phosphorylation of 1-beta-D-arabinofuranosylcytosine (araC) and 2', 2' difluorodeoxycytidine (dFdC) to their diphosphate forms catalyzed by human UMP CMP kinase. We cloned the cDNA of this enzyme to enable characterization of the recombinant protein, determine its expression in different tissues, and determine the chromosome location of the gene. We showed that the recombinant UMP-CMP kinase phosphorylated CMP, dCMP, and UMP with highest efficiency and dUMP, AMP, and dAMP with lower efficiency. The monophosphates of araC and dFdC were shown to be phosphorylated with similar efficiency as dCMP and CMP. We further showed, in a combined enzymatic assay, that human deoxycytidine kinase and UMP-CMP kinase together phosphorylated araC, dFdC, and 2',3'-dideoxycytidine to their diphosphate forms. Northern blot analysis showed that the UMP-CMP kinase mRNA was ubiquitously present in human tissues as a 3.9-kb transcript with highest levels in pancreas, skeletal muscle, and liver. The human UMP-CMP kinase gene was localized to chromosome 1p34.1-1p33 by radiation hybrid analysis. We further expressed the UMP-CMP kinase as a fusion protein to the green fluorescent protein in Chinese hamster ovary cells, and showed that the fusion protein was located in the cytosol and nucleus. PMID- 10462545 TI - The antiviral nucleotide analogs cidofovir and adefovir are novel substrates for human and rat renal organic anion transporter 1. AB - Nephrotoxicity is the dose-limiting clinical adverse effect of cidofovir and adefovir, two potent antiviral therapeutics. Because renal uptake likely plays a role in the etiology of cidofovir- and adefovir-associated nephrotoxicity, we attempted to identify a renal transporter capable of interacting with these therapeutics. A cDNA clone was isolated from a human renal library and designated human organic anion transporter 1 (hOAT1). Northern analysis detected a specific 2.5-kilobase pair hOAT1 transcript only in human kidney. However, reverse transcription-polymerase chain reaction revealed hOAT1 expression in human brain and skeletal muscle, as well. Immunoblot analysis of human kidney cortex demonstrated that hOAT1 is an 80- to 90-kilodalton heterogeneous protein modified by abundant N-glycosylation. Xenopus laevis oocytes expressing hOAT1 supported probenecid-sensitive uptake of [(3)H]p-aminohippurate (K(m) = 4 microM), which was trans-stimulated in oocytes preloaded with glutarate. Importantly, both hOAT1 and rat renal organic anion transporter 1 (rROAT1) mediated saturable, probenecid sensitive uptake of cidofovir, adefovir, and other nucleoside phosphonate antivirals. The affinity of hOAT1 toward cidofovir and adefovir (K(m) = 46 and 30 microM, respectively) was 5- to 9-fold higher compared with rROAT1 (K(m) = 238 and 270 microM, respectively). These data indicate that hOAT1 may significantly contribute to the accumulation of cidofovir and adefovir in renal proximal tubules and, thus, play an active role in the mechanism of nephrotoxicity associated with these antiviral therapeutics. PMID- 10462546 TI - The cloned rat vanilloid receptor VR1 mediates both R-type binding and C-type calcium response in dorsal root ganglion neurons. AB - [(3)H]Resiniferatoxin (RTX) binding and calcium uptake by rat dorsal root ganglion (DRG) neurons show distinct structure-activity relations, suggestive of independent vanilloid receptor (VR) subtypes. We have now characterized ligand binding to rat VR1 expressed in human embryonic kidney (HEK293) and Chinese hamster ovary (CHO) cells and compared the structure-activity relations with those for calcium mobilization. Human embryonic kidney cells (HEK293/VR1 cells) and Chinese hamster ovary cells transfected with VR1 (CHO/VR1 cells) bound [(3)H]RTX with affinities of 84 and 103 pM, respectively, and positive cooperativity (Hill numbers were 2.1 and 1.8). These parameters are similar to those determined with rat DRG membranes expressing native VRs (a K(d) of 70 pM and a Hill number of 1.7). The typical vanilloid agonists olvanil and capsaicin inhibited [(3)H]RTX binding to HEK293/VR1 cells with K(i) values of 0.4 and 4.0 microM, respectively. The corresponding values in DRG membranes were 0.3 and 2.5 microM. HEK293/VR1 cells and DRG membranes also recognized the novel vanilloids isovelleral and scutigeral with similar K(i) values (18 and 20 microM in HEK293/VR1 cells; 24 and 21 microM in DRGs). The competitive vanilloid receptor antagonist capsazepine inhibited [(3)H]RTX binding to HEK293/VR1 cells with a K(i) value of 6.2 microM and binding to DRG membranes with a K(i) value of 8.6 microM. RTX and capsaicin induced calcium mobilization in HEK293/VR1 cells with EC(50) values of 4.1 and 82 nM, respectively. Thus, the relative potencies of RTX (more potent for binding) and capsaicin (more potent for calcium mobilization) are similar in DRG neurons and cells transfected with VR1. We conclude that VR1 can account for both the ligand binding and calcium uptake observed in rat DRG neurons. PMID- 10462547 TI - Oxidative inactivation of cytochrome P-450 1A (CYP1A) stimulated by 3,3',4,4' tetrachlorobiphenyl: production of reactive oxygen by vertebrate CYP1As. AB - Microsomal cytochrome P-450 1A (CYP1A) in a vertebrate model (the teleost fish scup) is inactivated by the aryl hydrocarbon receptor agonist 3,3',4,4' tetrachlorobiphenyl (TCB). Here, the mechanism of CYP1A inactivation and its relationship to reactive oxygen species (ROS) formation were examined by using liver microsomes from scup and rat and expressed human CYP1As. In vitro inactivation of scup CYP1A activity 7-ethoxyresorufin O-deethylation by TCB was time dependent, NADPH dependent, oxygen dependent, and irreversible. TCB increased microsomal NADPH oxidation rates, and CYP1A inactivation was lessened by adding cytochrome c. CYP1A inactivation was accompanied by loss of spectral P 450, a variable loss of heme and a variable appearance of P-420. Rates of scup liver microsomal metabolism of TCB were < 0.5 pmol/min/mg, 25-fold less than the rate of P-450 loss. Non-heme iron chelators, antioxidant enzymes, and ROS scavengers had no influence on inactivation. Inactivation was accelerated by H(2)O(2) and azide but not by hydroxylamine or aminotriazole. TCB also inactivated rat liver microsomal CYP1A, apparently CYP1A1. Adding TCB to scup or rat liver microsomes containing induced levels of CYP1A, but not control microsomes, stimulated formation of ROS; formation rates correlated with native CYP1A1 content. TCB stimulated ROS formation by baculovirus-expressed human CYP1A1 but not CYP1A2. The results indicate that TCB uncouples the catalytic cycle of CYP1A, ostensibly CYP1A1, resulting in formation of ROS within the active site. These ROS may inactivate CYP1A or escape from the enzyme. ROS formed by CYP1A1 may contribute to the toxicity of planar halogenated aromatic hydrocarbons. PMID- 10462548 TI - Incorporation of the pi subunit into functional gamma-aminobutyric Acid(A) receptors. AB - mRNA encoding the recently cloned gamma-aminobuytyric acid(A) receptor (GABAR) pi subunit is expressed in the hippocampus and in several non-neuronal tissues including the uterus and ovaries. Whereas native GABARs are pentamers composed primarily of alphabetagamma, alphabetadelta, or alphabetaepsilon subunits, it has not been demonstrated clearly that the pi subunit incorporates into functional GABARs to form alphabetapi receptors and, if so, with what properties. We provide electrophysiological evidence that the pi subunit can coassemble with either alpha5beta3 or alpha5beta3gamma3 subunits to produce recombinant GABARs with distinct pharmacological and biophysical properties. Compared with alpha5beta3 receptors, GABARs produced by coexpression of alpha5beta3pi subunits had a lower GABA EC(50) value, were enhanced to a lesser extent by loreclezole, had different IC(50) values for pregnenolone sulfate and lanthanum, and were insensitive to benzodiazepines. Incorporation of both pi and gamma3 subunits into an alpha5beta3gamma3pi isoform was suggested by reduced enhancement by diazepam and a high zinc IC(50) value. Current-voltage relations for the alpha5beta3pi subunit combination outwardly rectified more than currents from alpha5beta3gamma3 but less than alpha5beta3 combination GABARs. Single-channel alpha5beta3 GABAR currents had a main conductance state of 15.2 picoSeimens (pS). Coexpression of the pi subunit with alpha5beta3 subtypes increased the conductance level to 23.8 pS, similar to the conductance level of alpha5beta3gamma3 GABARs (26.9 pS). We conclude that the pi subunit coassembles with alpha, beta, and gamma subunits to form functional alphabetapi or alphabetagammapi GABARs and, thus, could have a significant impact on the function of native GABARs expressed in the brain or non neuronal tissue. PMID- 10462549 TI - A mutation in the second transmembrane region of the CB1 receptor selectively disrupts G protein signaling and prevents receptor internalization. AB - We mutated a conserved aspartate in the second transmembrane domain of the cannabinoid CB(1) receptor to asparagine (D164N), stably transfected it into AtT20 cells, and examined the coupling of this mutant receptor to several intracellular effectors that are targets of wild-type CB(1) receptor activation. We found that the D164N receptor binds the CB(1) agonist WIN 55,212-2 with an affinity matching that of the wild-type CB(1) receptor and inhibits Ca(2+) currents and cAMP production with an equivalent potency and efficacy. This mutation, however, blocks coupling of the receptor to the potentiation of inwardly rectifying potassium channel (KIR) currents and prevents internalization of the receptor after exposure to agonist. Although the mutant receptor did not internalize, we found it was still capable of activating p42/44 MAP kinase. In addition, we made a reciprocal mutation that exchanged the aspartate with an asparagine in the seventh transmembrane region (D164N/N394D). In other seven membrane-spanning receptors, this reciprocal mutation is known to restore functions disrupted by the mutation of the single conserved aspartate. However, activation of D164N/N394D did not potentiate KIR current, nor did it internalize. We conclude that D164 is necessary for potentiation of KIR current and internalization of receptor but not necessary for agonist binding, inhibition of cAMP production, inhibition of Ca(2+) currents, or activation of p42/44 MAP kinase. Furthermore, CB(1) receptor internalization is not necessary for MAP kinase activation. PMID- 10462550 TI - N-methyl-D-aspartate excitotoxicity: relationships among plasma membrane potential, Na(+)/Ca(2+) exchange, mitochondrial Ca(2+) overload, and cytoplasmic concentrations of Ca(2+), H(+), and K(+). AB - A high cytoplasmic Na(+) concentration may contribute to N-methyl-D-aspartate (NMDA)-induced excitotoxicity by promoting Ca(2+) influx via reverse operation of the Na(+)/Ca(2+) exchanger (NaCaX), but may simultaneously decrease the electrochemical Ca(2+) driving force by depolarizing the plasma membrane (PM). Digital fluorescence microscopy was used to compare the effects of Na(+) versus ions that do not support the NaCaX operation, i.e., N-methyl-D-glucamine(+) or Li(+), on: PM potential; cytoplasmic concentrations of Ca(2+), H(+), and K(+); mitochondrial Ca(2+) storage; and viability of primary cultures of cerebellar granule cells exposed to NMDA receptor agonists. In the presence of Na(+) or Li(+), NMDA depolarized the PM and decreased cytoplasmic pH (pH(C)); in the presence of Li(+), Ca(2+) influx was reduced, mitochondrial Ca(2+) overload did not occur, and the cytoplasm became more acidified than in the presence of Na(+). In the presence of N-methyl-D-glucamine(+), NMDA instantly hyperpolarized the PM, but further changes in PM potential and pH(C) were Ca-dependent. In the absence of Ca(2+), hyperpolarization persisted, pH(C) was decreasing very slowly, K(+) was retained in the cytoplasm, and cerebellar granule cells survived the challenge; in the presence of Ca(2+), pH(C) dropped rapidly, the K(+) concentration gradient across the PM began to collapse as the PM began to depolarize, and Ca(2+) influx and excitotoxicity greatly increased. These results indicate that the dominant, very likely excitotoxic, component of NMDA-induced Ca(2+) influx is mediated by reverse NaCaX and that direct Ca(2+) influx via NMDA channels is curtailed by Na-dependent PM depolarization. PMID- 10462551 TI - Molecular modeling of the intrastrand guanine-guanine DNA adducts produced by cisplatin and oxaliplatin. AB - Intrastrand DNA adducts formed by cisplatin and oxaliplatin were modeled with molecular mechanics minimization and restrained molecular dynamics simulations in a comparative study. A reasonable set of force field parameters for the Pt atom were refined by using the available cisplatinated DNA crystal structure as a guide. This crystal structure was also used as the starting structure for the simulations. Analysis of the resulting structures indicated that the covalent effects of oxaliplatin coordination on DNA structure were very similar to those of cisplatin. The most prominent difference between the two structures resulted from the presence of the 1,2-diaminocyclohexane ring in the oxaliplatin adduct. The modeling indicated that this ring protrudes directly outward into, and fills much of, the narrowed major groove of the bound DNA, forming a markedly altered and less polar major groove in the area of the adduct. The differences in the structure of the adducts produced by cisplatin and oxaliplatin are consistent with the observation that they are differentially recognized by the DNA mismatch repair system. PMID- 10462552 TI - The N terminus domain of type VI adenylyl cyclase mediates its inhibition by protein kinase C. AB - Previous results from our laboratory have shown that phosphorylation of type VI adenylyl cyclase (ACVI) by protein kinase C (PKC) caused suppression of adenylyl cyclase activity. In the present study, we investigated the role of the N terminus cytosolic domain of ACVI in this PKC-mediated inhibition of ACVI. Removal of amino acids 1 to 86 of ACVI or mutation of Ser(10) (a potential PKC phosphorylation site) into alanine significantly relieved the PKC-mediated inhibition and markedly reduced the PKC-evoked protein phosphorylation. PKC also effectively phosphorylated a recombinant N terminus cytosolic domain (amino acids 1-160) protein of ACVI and a synthetic peptide representing Ser(10). In addition, the amino acids 1 to 86 truncated mutant exhibited kinetic properties similar to those of the wild type. Taken together, these data demonstrate that the highly variable N terminus cytoplasmic domain of ACVI is a regulatory domain with a critical role in PKC-mediated suppression, which is a hallmark of this adenylyl cyclase isozyme. In addition, Ser(10) was found to serve as an acceptor for the PKC-mediated phosphorylating transfer of ACVI. PMID- 10462553 TI - Influence of G protein type on agonist efficacy. AB - An agonist at a specific G protein-coupled receptor may exhibit a range of efficacies for any given response in a cell-specific manner. We report that the relationship between different states of agonism is regulated by the type of G protein expressed in the cell. In NIH-3T3 alpha(2)-adrenergic receptor (AR) transfectants, the alpha(2)-AR agonists clonidine, oxymetazoline, UK-14304, and epinephrine increased [(35)S]guanosine-5'-O-(3-thio)triphosphate binding in a dose-dependent manner from a basal value of 101.2 +/- 6. 5 fmol/mg to a maximal response (100 microM) of 196.6 +/- 9.8, 182.3 +/- 2, 328.1 +/- 11.2, and 340.6 +/ 3 fmol/mg, respectively. Thus, clonidine and oxymetazoline behaved as partial agonists. Receptor-mediated activation of G proteins in membrane preparations was blocked by cell pretreatment with pertussis toxin, indicating receptor coupling to the subgroup of pertussis toxin-sensitive G protein (Gialpha2,3) expressed in NIH-3T3 cells. Ectopic expression of Goalpha1 but not Gialpha1 increased the relative efficacy of clonidine and oxymetazoline such that the two ligands now behaved as close to full agonists in this assay system. The relationship between full and partial agonists in the different genetic backgrounds was not altered by progressive reduction in the amount of G protein available for coupling to receptor. The increased efficacy observed for clonidine in the Goalpha1 transfectants was not due to changes in the relative affinities or amounts of high-affinity, Gpp(NH)p-sensitive binding of agonist. These data suggest that there is little difference in the ability of clonidine to interact with or stabilize alpha(2)-AR-Gialpha2/Gialpha3 versus alpha(2)-AR-Goalpha1 complexes, but that the subsequent step of signal transfer from receptor to G protein is more readily achieved for the clonidine/alpha(2)-AR/Goalpha1 complex. Such observations have important implications for receptor theory and drug development. PMID- 10462555 TI - Xenograft Transplantation: Knowledge of Consequence? PMID- 10462554 TI - Identification, molecular cloning, expression, and characterization of a cysteinyl leukotriene receptor. AB - The cysteinyl leukotrienes (CysLTs) have been implicated in the pathophysiology of inflammatory disorders, in particular asthma, for which the CysLT receptor antagonists pranlukast, zafirlukast, and montelukast, have been introduced recently as novel therapeutics. Here we report on the molecular cloning, expression, localization, and pharmacological characterization of a CysLT receptor (CysLTR), which was identified by ligand fishing of orphan seven transmembrane-spanning, G protein-coupled receptors. This receptor, expressed in human embryonic kidney (HEK)-293 cells responded selectively to the individual CysLTs, LTC(4), LTD(4), or LTE(4), with a calcium mobilization response; the rank order potency was LTD(4) (EC(50) = 2.5 nM) > LTC(4) (EC(50) = 24 nM) > LTE(4) (EC(50) = 240 nM). Evidence was provided that LTE(4) is a partial agonist at this receptor. [(3)H]LTD(4) binding and LTD(4)-induced calcium mobilization in HEK-293 cells expressing the CysLT receptor were potently inhibited by the structurally distinct CysLTR antagonists pranlukast, montelukast, zafirlukast, and pobilukast; the rank order potency was pranlukast = zafirlukast > montelukast > pobilukast. LTD(4)-induced calcium mobilization in HEK-293 cells expressing the CysLT receptor was not affected by pertussis toxin, and the signal appears to be the result of the release from intracellular stores. Localization studies indicate the expression of this receptor in several tissues, including human lung, human bronchus, and human peripheral blood leukocytes. The discovery of this receptor, which has characteristics of the purported CysLT(1) receptor subtype, should assist in the elucidation of the pathophysiological roles of the CysLTs and in the identification of additional receptor subtypes. PMID- 10462556 TI - Molecular Analysis in a Patient With Waldenstrom's Macroglobulinemia Reveals a Rare Case of Biclonality. AB - Background: The immunoglobulin and T-cell receptor gene rearrangement test is used to identify monoclonal populations in B- and T-cell malignancies and has become an important adjunct to morphologic analysis and immunophenotyping by flow cytometry. Waldenstrom's macroglobulinemia (WM) is typically a monoclonal proliferation of B cells with morphology of plasmacytoid lymphocytes and production of monoclonal IgM. Methods and Results: We report a case of WM with biclonal gammopathy (IgM kappa and IgM lambda) involving the blood and a diffuse lymphoplasmacytic infiltrate in the bone marrow in an 83-year-old man. Immunophenotyping of the blood and bone marrow aspirate revealed B cells expressing IgM lambda surface immunoglobulins and CD5+, CD19+, and CD20+ surface markers. Gene-rearrangement analysis with the Southern blot technique revealed multiple rearranged bands in each lane of restricted patient DNA after probing with both immunoglobulin heavy (J(H))- and light (J(kappa))-chain gene probes. Conclusions: Biclonal gammopathy in WM and biclonal B-cell proliferations as determined by gene-rearrangement studies are rare entities, and few evaluations of them are reported in the literature. To our knowledge, this case is the first one of biclonal WM to have been evaluated by serum protein immunofixation, immunohistologic staining, immunophenotyping by flow cytometry, and immunoglobulin gene-rearrangement analysis. PMID- 10462557 TI - Semiquantitative Detection of Abnormal CD44 Transcripts in Colon Carcinomas by Reverse Transcription-Polymerase Chain Reaction Enzyme-linked Immunosorbant Assay (RT-PCR ELISA). AB - Background: Abnormal expression of CD44 variant RNA has been detected in a variety of human tumors and has been shown to be a potential diagnostic marker. To date, such analysis requires time-consuming gel electrophoresis, blotting, and autoradiographic procedures, and this approach may not be suitable for routine laboratory examinations. We have developed a rapid and semiquantitative reverse transcription-polymerase chain reaction enzyme-linked immunosorbent assay (RT-PCR ELISA) method and used it to analyze CD44 expression in colon carcinoma tissues and exfoliated cancer cells in colon luminal washings. Methods and Results: RNA was extracted from sample cells and tissues and converted to cDNA. PCR amplification products, labeled by incorporation of digoxigenin-11-dUTP, were hybridized with biotinylated probes complementary to CD44 exon 12 or to exons in the standard portion (CD44s) of the gene. Hybridized DNA complexes were immobilized on streptavidin-coated microtiter plates, and the bound PCR products were detected with a peroxidase-conjugated antibody to digoxigenin. CD44-derived PCR products were quantified by absorbance of a chromogenic reaction. Elevated expression of CD44 variant exon 12 was detected initially by Southern blot analysis in all of the 9 colon carcinoma tissues, while weak expression was observed in only 3 of 9 normal mucosas. This tumor-related differential expression was confirmed by the newly developed PCR-ELISA method. Elevated expression of CD44 exon 12 was also detected in exfoliated colonic epithelial cells from 10 of 13 carcinoma cases but not in exfoliated cells from 4 patients with inflammatory bowel disease. Conclusions: Raised expression of CD44 variant exon transcripts can be detected reliably in colonic tumor tissue and in exfoliated colonic cancer cells by a semiquantitative RT-PCR ELISA method. This was shown to be as sensitive as conventional RT-PCR using chemiluminescent detection. Therefore, CD44-based RT-PCR ELISA could facilitate detection of neoplasia in clinical specimens including colon washings and naturally micturated urine. PMID- 10462558 TI - Accumulation of Immature Intron-containing CD44 Gene Transcripts in Breast Cancer Tissues. AB - Background: Disorderly expression of the CD44 gene is a characteristic feature of many common types of human malignancy. The explanation for the diversity, abundancy, and abnormally large size of the resulting transcripts is under investigation. Methods and Results: This study used reverse transcription polymerase chain reaction (RT-PCR)/Southern blot hybridization to examine the expression of the CD44 gene in fresh tissue samples from 21 breast cancers and 11 matched non-neoplastic breast tissue specimens from the same patients. Using probes for several exons and for a noncoding region (intron 9) of the gene, it was found that the previously described elevated levels of CD44 transcripts in malignant tumors of this organ include many unusual, alternatively spliced isotypes as well as immature forms that retain this intron and probably several others. All of the exons that were tested were involved in the disorderly overexpression of this gene observed in all of the cancer tissues, and we were able to detect retention of the intron 9 sequence in 14 (67%) of the 21 tumor samples. In contrast, it was possible to detect signals in only 3 (27%) of the 11 samples from nonmalignant breast tissue with this probe, and these were extremely faint. Conclusions: These findings imply that there is a profound disorder in the regulation of production, splicing, and processing of CD44 pre-mRNA in breast cancer tissues comparable to that described in tumors of many other organs. The clinical implication of this information is that analysis of tissue samples containing borderline or suspected premalignant lesions for the presence of these molecular abnormalities, which appear to be characteristic of neoplasia, may in due course help assessment of individual patient prognosis and optimization of treatment. PMID- 10462559 TI - Microchip-based Devices for Molecular Diagnosis of Genetic Diseases. AB - Microchips, constructed with a variety of microfabrication technologies (photolithography, micropatterning, microjet printing, light-directed chemical synthesis, laser stereochemical etching, and microcontact printing) are being applied to molecular biology. The new microchip-based analytical devices promise to solve the analytical problems faced by many molecular biologists (eg, contamination, low throughput, and high cost). They may revolutionize molecular biology and its application in clinical medicine, forensic science, and environmental monitoring. A typical biochemical analysis involves three main steps: (1) sample preparation, (2) biochemical reaction, and (3) detection (either separation or hybridization may be involved) accompanied by data acquisition and interpretation. The construction of a miniturized analyzer will therefore necessarily entail the miniaturization and integration of all three of these processes. The literature related to the miniaturization of these three processes indicates that the greatest emphasis so far is on the investigation and development of methods for the detection of nucleic acid, followed by the optimization of a biochemical reaction, such as the polymerase chain reaction. The first step involving sample preparation has received little attention. In this review the state of the art of, microchip-based, miniaturized analytical processes (eg, sample preparation, biochemical reaction, and detection of products) are outlined and the applications of microchip-based devices in the molecular diagnosis of genetic diseases are discussed. PMID- 10462560 TI - Importance of Molecular Diagnostics in the Identification and Control of Emerging Infections. PMID- 10462561 TI - Xenotransplantation: The Risk of Emerging Infections. PMID- 10462562 TI - Xenotransplantation and Possible Emerging Infectious Diseases. PMID- 10462563 TI - Xenotransplantation: Strategies to Achieve Donor-specific Tolerance and Immune Reconstitution Across Species Barriers Through Mixed Bone Marrow Chimerism. AB - Background: Allogeneic organ donation will never provide the quantities of organs required for patients with end-stage disease. It is estimated that over 3,000 candidates for a potentially lifesaving sold-organ transplant will die this year while on the waiting list as a result of the donor shortage. A further understanding of the mechanisms of xenoreactivity might make xenotransplantation a viable therapeutic option. This article reviews the benefits of xenotransplantation, including species-specific disease resistance and the potential unlimited supply of donor organs. Methods and Results: Experimental results in rodents have demonstrated that bone marrow stem cells engraft in xenogeneic recipients. Lymphohematopoietic cells arising in a xenogeneic environment undergo normal maturation and negative selection of autoreactive clones. Recipients exhibit donor-specific tolerance to solid-organ and cellular xenografts, yet maintain normal reactivity to third-party antigens. Bone marrow transplantation as a means of developing mixed xenogeneic chimerism and donor specific tolerance is reviewed. Evidence that baboon cells are resistant to human immunodeficiency virus type 1 and hepatitis B virus is presented, and the rationale for transplantation of baboon hematopoietic stem cells to achieve immune reconstitution in late stage acquired immune deficiency syndrome is discussed. Conclusions: Donor-specific transplantation tolerance induced by mixed xenogeneic chimerism following bone marrow transplantation may provide a means for overcoming barriers to xenotransplantation. PMID- 10462564 TI - Immunologic Issues in Xenotransplantation. PMID- 10462565 TI - Beyond Rejection and Infection: The Challenge of Cell Biology to Xenotransplantation. PMID- 10462566 TI - Ethical Issues Associated With Xenotransplantation: The Case of Baboon Bone Marrow to Human Transplantation. PMID- 10462567 TI - Nucleic Acid-based Pathogen Discovery Techniques: Potential Application to Xenozoonoses. AB - Pathogen discovery techniques based on detection and characterization of microbial nucleic acids have potential application to the field of xenotransplantation. Techniques such as broad-range polymerase chain reaction and representational difference analysis may make it possible to define the zoonotic flora present within donor animal species to an extent never before possible. This review describes the current practice of these techniques, with an emphasis on the use of these methods for identifying zoonotic agents. Identification of potential pathogens in colony-reared donors may lead to isolation and selective breeding practices that could ultimately eliminate these agents from the donor pool. PMID- 10462568 TI - Strategies for Diagnosis of Xenotransplant-associated Retroviral Infections. AB - The demand for tissue for human-to-human transplantation consistently exceeds the available supply, resulting in attempts to use organs or tissues from animals; a field known as xenotransplantation. A major concern in xenotransplantation is the risk of infection of human recipients with new xenogeneic infectious agents and subsequent transmission of these infections to the general population. Of particular importance are infections with retroviruses that persist in the host and may allow silent human-to-human transmission. Pigs and baboons are considered as sources for human xenotransplants and are known to be infected with a number of retroviruses. Adequate diagnosis of pig and baboon retrovirus infections is required for pretransplant screening of animal sources to minimize risks of transmission to recipients and also for monitoring post-transplant infections in the human recipients. Strategies for serologic, molecular, and virologic diagnosis are described. Examples of virus-specific methods for detection of known retroviruses are also shown. Emphasis is made on the need for baboon and pig cell-specific polymerase chain reaction assays that are necessary for interpretation of polymerase chain reaction results of endogenous retroviruses. Generic methods for detection of novel or variant retrovirus are described and include screening by Amp-RT, an ultrasensitive reverse transcriptase assay. PMID- 10462569 TI - Rapid Identification and Differentiation of Baboon Alphaherpesvirus SA8 From Closely Related HSV-1 and HSV-2 in Xenograft Transplant Recipients. AB - Background: Rapid differential diagnosis of SA8 infection in humans following xenograft transplantation is an essential part of organ cell donor recipients. Methods and Results: We have developed a polymerase chain reaction (PCR)-based test using synthetic oligonucleotide primers and probe to detect SA8 DNA in clinical samples. After assessing the specificity of the PCR for detection of laboratory SA8 isolates, the method was used in the investigation of human and baboon specimens and compared to virus culture techniques. Conclusions: The PCR appears to be more sensitive than conventional viral isolation and thus can be used for rapid identification of SA8 virus in cases when conventional virus cultures may not be useful. Further, the PCR can be used to unequivocally distinguish closely related human viruses HSV-1 and HSV-2 from SA8. PMID- 10462571 TI - Certification for Molecular Diagnostics Laboratory Directors and Personnel. PMID- 10462570 TI - PCR Identification and Differentiation of Baboon Cytomegalovirus From Other Human and Nonhuman Primate Cytomegalovirus. AB - Background: Differential identification of baboon alpha, beta, and gamma herpesviruses is an essential technology in order to monitor xenozoonotic transmission of baboon viruses to foreign species organ and/or cell recipients. We present polymerase chain reaction techniques that will differentiate known baboon cytomegaloviruses (CMV) from their closely related counterparts found in humans. Methods and Results: Polymerase chain reaction techniques for identification of the known beta herpesviruses commonly present in the baboon are reported. The techniques described also permit the unequivocal differentiation of these virus types from closely related human as well as other nonhuman primate viruses in the family herpesviridae. Methods are based upon sequence analysis of specifically selected genes of baboon CMV. Primer pairs from sequence analyses were selected based upon nonhomologous sequences in gene homologues of human CMV. Unique, baboon species specific amplimers were identified both with ethidium bromide staining and with radiolabeled probe analyses. Conclusions: With the described techniques, it is possible to monitor organ/cell recipients of xenograft transplantation for the establishment of baboon CMV in the foreign human host. Monitoring can be performed throughout the life of the recipient in effort to rule out host susceptibility to baboon CMV, as well as to rule out potential host:donor recombinant viruses formed between the closely related members of herpesvirus family. PMID- 10462573 TI - Molecular Diagnosis-One Year Old! PMID- 10462572 TI - Regulation of Analyte-specific Reagents. PMID- 10462574 TI - Identification of MYCN Copy Number Heterogeneity by Direct FISH Analysis of Neuroblastoma Preparations. AB - Background: Amplification of MYCN and deletion of 1p in neuroblastoma are associated wtih a poor prognosis. MYCN copy numbers of 10 or greater are considered to be indicative of poor outcome. However, most molecular genetic methods for estimating the number of MYCN genes do not directly address copy number heterogeneity at the cellular level. Methods and Results: MYCN copy number was assessed by fluorescence in situ hybridization (FISH), differential polymerase chain reaction (PCR), and Southern blot analysis, using 29 patient tumors. Copy number estimates by PCR or Southern blot analyses identified MYCN amplification in 11 tumors. There was complete concordance between FISH MYCN results in all 11 tumors with amplification. FISH identified 5 tumors with marked heterogeneity for MYCN copy number. Two tumors in which a small percentage of cells within the specimen were amplified would have gone undetected by molecular genetic methods alone. Conclusions: FISH offers the advantage over the other methods of detecting heterogeneity, thereby revealing tumors with small numbers of amplified cells that would otherwise be missed and, in cases of low (3-10) copies of MYCN, distinguishing small numbers of amplified cells (poor prognosis) from triploid tumors (good prognosis). FISH also allows detection of 1p deletion using the same preparations, which represents another advantage. A combination of FISH and conventional molecular methods provides an accurate definition of sample heterogeneity, and should be routinely applied to all neuroblastomas with low levels of MYCN copy number. PMID- 10462575 TI - Combined In Situ Hybridization and Immunohistochemistry for Automated Detection of Cytomegalovirus and p53. AB - Background: Cytomegalovirus (CMV) infection has been shown to be associated with p53 overexpression in coronary artery restenosis. We investigated the occurance of this association in other forms of CMV infection using an automated in situ hybridization (ISH) technique. Methods and Results: We performed ISH for CMV using digoxigenin-labeled or biotinylated probes followed by avidin-alkaline phosphatase and nitroblue tetrazolium color substrate. Immunohistochemistry (IHC) was then performed using an anti-p53 antibody utilizing streptavidin immunoperoxidase and 3,3;-diaminobenzidine tetrahydrochloride as a chromogen. Sixteen cases with characteristic cytomegalic inclusions from a variety of body sites were examined. All 16 cases were positive for CMV by ISH. Nine of sixteen expressed nuclear p53. Six of these nine cases showed viral cytopathic effect in the cells with p53 expression. In an illustrative case, double colocalized staining for CMV and p53 protein was demonstrated in individual cytopathic cells. When microwave antigen retrieval was necessary, ISH was performed before IHC, and our standard microwaving time was reduced by two-thirds. Conclusions: The colocalization of p53 protein overexpression with CMV within single cells adds further evidence that this overexpression is a viral-induced phenomenon. The combined ISH and IHC assay can be carried out in a rapid automated mode, increasing the ease of investigating relationships between message and protein expression within single cells in a wide variety of settings. PMID- 10462576 TI - Resistance to Activated Protein C: Comparison of Three Different PCR Methods for Detection FV R506Q. AB - Background: Resistance to activated protein C(APC) is the most prevalent identifiable risk factor for inherited thrombophilia. Over 90% of APC resistance results from a single point mutation in the Factor V gene. The mutation, termed FV R506Q of FV Leiden, predicts an abnormal Factor Va protein in which arginine, at amino acid position 506, is replaced by glutamine, rendering Factor Va resistant to proteolytic inactivation by APC, thus establishing a life long hypercoagulable state. The current study compared three different polymerase chain reaction (PCR)-based approaches for the detection of FV R506Q. Methods and Results: Sixty-seven patient blood samples were genotyped by (1) analyzing for loss of a Mnl I recognition site, an acquired restriction-fragment-length polymorphism (RFLP) due to FV R506Q; (2) primer-engineered RFLP wherein the presence of FV R506Q results in generation of a novel Nla III recognition site; and (3) allele-specific PCR. Sixty-five of 67 patient samples yielded concordant genotype results by all three PCR methods. Of the remaining 2 of the 67 patients, a "nondiagnostic" result was obtained for either allele-specific PCR or primer engineered RFLP. Conclusions: A comparative analysis of 67 patient samples demonstrated that primer engineered RFLP and allele-specific PCR offer feasible alternative or confirmatory testing approaches to Mnl I RFLP for the detection of FV R506Q. A high degree of diagnostic concordance was observed for all three methods, and no false positive or negative results were observed with the Mnl I RFLP technique. PMID- 10462577 TI - Detection of BCRabl in Acute Leukemia by Molecular and Cytogenetic Methods. AB - Background: The Philadelphia chromosome (Ph), t(9;22)(q34;q11), is detected by karyotyping in a minority of patients with acute leukemia. Ph results in fusion of the c-abl oncogene on chromosome 9 with the breakpoint cluster region BCR gene on chromosome 22. The purpose of this study was to compare reverse trascriptase polymerase chain reaction (RT-PCR) for BCR/abl fusion to cytogenetic methods for Ph detection in patients with acute leukemia. Methods and Results: Peripheral blood and bone marrow samples from cases of adult acute myelogenous leukemia (AML) and acute lymphoblastic leukemia (ALL) were examined for Ph by RT-PCR, karyotyping, and fluorescence in situ hybridization (FISH). Using total cellular RNA and a single primer pair, cDNA was transcribed, amplified, electorphoresed, and probed for BCR/abl fusion. Patient cells and SUPB15 and K562 cell lines were used as breakpoint controls. Karyotyping was done by standard Giemsa banding. FISH was performed on bone marrow smears using digxigenin-labeled DNA probes for major and minor bcr breakpoints (corresponding to involvement of major bcr exons 2/3 and minor bcr exon 1, respectively) and biotin-labeled DNA probes for abl. Rhodamine-conjugated antidigoxigenin and fluorescein-conjugated avidin yielded red and green fluorescent signals, respectively. A total of 32 samples from patients with AML were studied, 20 from patients with de novo or relapsed AML and 12 from patients in remission. Five of 32 cases of AML (16%) were RT-PCR+/Ph+ all with major bcr breakpoints between exons 3 and 4. One of the 32 cases (3%) was RT PCR+/Ph+; this case was the only positive remission sample. Of the four T-PCR+/Ph cases, one showed t(2;17), one showed t(9;11), and two had a normal karyotype. FISH was done in three RT-PCR+ cases, yielding positive results with the major probe in two. A total of 22 samples from patients with ALL were studies, 15 from patients with de novo or relapsed ALL and seven from patient remission. Seven of 22 cases of ALL (32%) were RT-PCR+, four with major-bcr breakpoints between exons 3 and 4, and three with breakpoints in the minor-bcr. Two of the 22 (9%) cases were RT-PCR+/Ph+. Of the five RT-PCR+/Ph- cases, two showed a 22q- but lacked the typical Ph break on chromosome 9, 1 showed a 12p-, and two had a normal karyotype. FISH was performed in four RT-PCR+ cases, yielding positive results with the major probe in two cases and with the minor probe in two cases. Conclusions: RT-PCR is more sensitive than karyotyping, detecting masked Ph or translocations not found by cytogenetic analysis. FISH is a helpful adjunctive test when used to confirm BCR/abl fusion in RT-PCR+/Ph- cases. PMID- 10462578 TI - Use of Myeloperoxidase mRNA in Monitoring Patients With Acute Myelogenous Leukemia for Early Detection of Circulating Blasts. AB - Background: Expression of the myeloperoxidase (MPO) gene is specific for myeloid precursors and their leukemic counterparts. Unlike the enzyme, MPO mRNA is found only in early myeloid precursors; this makes MPO mRNA a good marker for myeloid lineage of leukemic blasts. A reverse transcriptase-polymerase chain reaction (RT PCR) method for MPO mRNA detection was developed and used in the diagnosis of acute myelogenous leukemia (AML). In this study, we investigated the use of MPO mRNA for early detection of circulating blasts in patients with AML during and after chemotherapy. Methods and Results: MPO mRNA detection by RT-PCR was performed on cellular material from archival smears of preipheral blood (PB) and bone marrow aspirate from 16 patients previously diagnosed with AML, types MO-M5. MPO mRNA findings were correlated with morphology and flow-cytometric data. A group of six patients diagnosed with adult de novo acute lymphoblastic leukemia served as a negative patient control group for this retrospective study. MPO mRNA findings in PB appear to follow two patterns in patients with complete remission: (1) sustained positivity throughout the clinical course, correlated with relapse; and (2) initial positivity followed by sustained negativity, correlated with long complete remissions. For the only patient in this study found in partial remission, MP mRNA positivity in PB was seen throughout the clinical course. No MPO mRNA positivity was detected in the PB of acute lymphoblastic leukemia cases. Conclusions: A highly sensitive method for detection of MPO mRNA, such as RT-PCR, is useful in monitoring patients with AML for confirming or ruling out complete remission at the molecular level. The pattern of MPO mRNA positivity over time appears to be important and to correlate with clinical course, with sustained positivity being associated wtih impending relapse, while a switch from initial positivity to sustained negativity appears to be associated with long complete remssion. Studies of larger patient groups are necessary to confirm these initial findings. PMID- 10462579 TI - Exonuclease-released Fluorescence Detection of Human Parvovirus B19 DNA. AB - Background: The 5;-->3;-exonuclease activity of Thermus aquaticus (Taq) DNA polymerase permits polymerase chain reaction (PCR) product detection immediately after amplification using a fluorogenic probe. This approach eliminates the requirement for gel electrophoresis or enzyme immunoassays (EIA). The ligonucleotide probe is labeled with a reporter dye at its 5' terminus and a quencher dye at its 3' terminus and is present during DNA amplification. The exonuclease cleaves the reporter molecule from the probe-template hybrid, releasing it from the influence of the quencher molecule. The result is an increase in reporter fluorescence that can be read directly in a fluorescence spectormeter. In contrast to time-consuming gel electrophoresis and Southern blot hybridization or an EIA, this method can produce results from an entire 96-well microtiter plate in 15 minutes. Methods and Results: A B19-specific fluorogenic probe was synthesized containing a 5'-FAM label and a 3'-TAMRA label. Thirty clinical samples were analyzed for Human parvovirus B19 DNA by PCR amplification using both the fluorogenic and EIA method. Conclusions: Results generated with the fluorogenic probe correlated perfectly with those of the EIA, and the method would be particularly useful for high-volume work loads where gel or EIA-based approaches would be cumbersome. PMID- 10462580 TI - Toluidine Blue Dye-Binding Method for Measurement of Genomic DNA Extracted From Peripheral Blood Leukocytes. AB - Background: The toluidine blue DNA assay has characteristics that suggest its suitability for concentration determinations of genomic DNA extracted from peripheral blood leukocytes. These include visible wavelength readings, rapid analytical time, useful sensitivity, and low reagent costs. We validated this assay for genomic DNA and devised a simple method to identify extracts that exceed the linear range of the assay. Methods and Results: DNA was prepared from 109 blood samples by a salt extraction method. The toluidine blue assay was linear for DNA concentrations from 0 to 500 ng/uL. By performing bichromatic absorption readings at 570 and 628 nm, extracts with DNA concentrations exceeding the linear range could be identified up to at least 1300 ng/uL. Analytical precision and accuracy studies gave acceptable results. There was excellent agreement between DNA concentrations determined by the toluidine blue assay and ultraviolet spectrophotometry. RNA was confirmed to be significant positive interferent. The detection limit of genomic DNA was determined to be 15 ng/uL. Conclusions: The toluidine blue DNA assay is suitable for measuring DNA concentrations in extracts obtained from peripheral blood. This assay appears to be suitable for automation as a visible spectrophotometric DNA method. PMID- 10462581 TI - Molecular Assessment of Clonality in Lymphoproliferative Disorders: I. Immunoglobulin Gene Rearrangements. AB - The non-Hodgkins' lymphomas are a diverse group of malignancies with characteristic clinical and laboratory features. The routine evaluation of these lesions includes histologic and immunophenotypic analyses. In the small percentage of cases that pose diagnostic challenges, the algorithm can include molecular genetic studies to help render a diagnosis. Unique immunoglobulin heavy and light-chain or T-cell-receptor gene rearrangements are used to establish clonality in B- and T-cell proliferations. Whereas clonality is usually a hallmark of a malignant process, the absence of clonality does not exclude malignancy. This is predicted on the disparate biology of the various lympho proliferative processes and on the technical aspects of the tests used for their identification. As molecular genetic testing for gene rearrangements becomes commonplace in the clinical laboratory, the variety of methods for performing these tests has increased. Since molecular testing may be the deciding feature in making a diagnosis of malignancy, the purpose of this review is to discuss the advantages and problems of the polymerase chain reaction and Southern blot method in detecting immunoglobulin gene rearrangements in B cells. The added goal of this summary is to encourage the laboratorian to interpret these tests in the context of complete clinical and histologic data. PMID- 10462582 TI - Clinical News Update. PMID- 10462583 TI - Letter to the Editor. PMID- 10462584 TI - How Do I Know What I Think Till I See What I Say? PMID- 10462585 TI - Chromosome 8 Losses in Colorectal Carcinoma: Localization and Correlation With Invasive Disease. AB - Background: Allelic losses from the short arm of chromosome 8 (8p) are frequent in a variety of epithelial malignancies. In colorectal cancer, there are two discrete regions of 8p loss of hterozygosity (LOH), suggesting the existence of two putative tumor suppressor genes. Previous studies have shown an association of 8p LOH with tumor invasiveness. To better define the deletion extent and the clinical significance of these losses, a series of 41 colorectal cancers were examined for 8p LOH and correlated with clinical features. Methods and Results: Paired normal and enriched tumor DNA from the same individual was typed by polymerase chain reaction for 11 microsatellite polymorphisms and scored as positive or negative for 8p LOH. Loss of 8p markers was observed in 44% of the cases. Most cases had large deletions, but several had localized interstitial losses, enabling specification of two nonoverlapping regions of LOH. The telomeric region of loss is defined by the markers D8S262 and D8S133 at 8p22, and the centromeric region is proximal to NEFL. Clinical, histopathologic, and molecular data were obtained and a significant correlation of 8p LOH with microinvasion (invasion of lymphatics, vessels, or perineurium, ascertained by light microscopy) (P=.01) and also with loss of chromosome arm 18q (P=.001) was found. Conclusions: An association of 8p allelic loss with poor outcome was demonstrated. The correlation between 8p loss and 18q loss suggests that 8p LOH is a late event in the multistep model of colorectal carcinogenesis. 8p LOH may provide a clinically useful prognostic marker in colorectal cancer, thereby warranting further testing. The involvement of two independent loci on 8p is confirmed, and the refined localization of these sites will contribute to the eventual identification of these genes, which appear to play an important role in the progression of epithelial malignancies. PMID- 10462586 TI - Presence of Human Herpesvirus 8 Is Specific to Kaposi's Sarcoma Lesion. AB - Background: Human herpesvirus 8(HHV8) has previously been shown to be associated with Kaposi's sarcoma (KS) skin lesions in HIV-positive and negative patients. This study was designed to test, on a microscopic level, using the microdissection technique in frozen and formalin-fixed tissue, whether HHV8 DNA is specific to the KS lesion. Methods and Results: Thirty-seven evaluable KS lesions were microdissected and DNA extracted, and HHV8-specific polymerase chain reaction (PCR) was performed. All of the KS lesions showed a PCR product; none of the microscopically adjacent normal tissue samples showed a PCR product. One of the PCR products was sequenced, confirming the presence of HHV8 DNA. Conclusions: HHV8 was shown to be specific to the KS lesion at the microscopic level. PMID- 10462588 TI - Editorial Comment. PMID- 10462587 TI - Modern Diagnostic Methods in Ewing's Sarcoma Family: Six Patients With Histologic Soft Tissue Tumors. AB - Background: Soft tissue tumors often present a major diagnostic challenge for the pathologist. The correct diagnosis has important prognostic and therapeutic consequences. In recent years significant progress has been made in identifying characteristic chromosomal abnormalities associated with certain solid tumors. More than 85% of tumors in the Ewing's sarcoma (ES) family contain a specific t(11;22) (q24;q12) translocation. Methods and Results: We present six patients with a soft tissue tumor of which only four were diagnosed primarily as belonging to the ES family. All cases were further examined by the following methods: immunohistochemistry with MIC2, cytogenetics, nested reverse transcription polymerase chain reaction of the t(11;22), using fresh-frozen or formalin-fixed, paraffin-embedded archival material. Conclusions: This method clearly allowed the diagnosis of a tumor of the ES family in all six cases. PMID- 10462589 TI - Diagnostic Detection of Herpes Simplex and Hepatitis C Viral Amplicons by Capillary Electrophoresis: Comparison With Southern Blot Detection. AB - Background: As a result of the large number of DNA-based clinical assays, there is intense interest in making polymerase chain reaction (PCR)-amplified DNA product analysis faster, more cost-effective, and more automated. Methods and Results: In this study, an evaluation of the use of capillary gel electrophoresis with laser-induced fluorescence detection is described as a means of analyzing postamplification PCR products from clinical specimens. Sixty-six herpes simplex virus and 152 hepatitis C virus amplicons were analyzed after PCR and reverse transcription PCR, respectively. It is shown that the use of a physical gel buffer system in a short capillary in conjunction with laser-induced fluorescence detection allows for sensitive detection of herpes simplex virus- and hepatitis C virus-specific DNA fragments in an expedient manner. Interinstrument and intercapillary reproducibility of the migration time was evaluated and found to be excellent. The advantages and disadvantages over agarose gel electorphoresis Southern blot analysis are summarized. Conclusions: The advantages offered by capillary gel electrophoresis with laser-induced fluorescent detection including rapid and sensitive analysis, ease of setup, reduced cost, and possibility for automation, make this procedure a viable alternative to more labor-intensive agarose gel electrophoresis-Southern blot analysis as molecular diagnostic methodology. PMID- 10462591 TI - Detection of Septata intestinalis (Microsporidia) Cali et al. 1993 Using Polymerase Chain Reaction Primers Targeting the Small Submit Subunit Ribosomal RNA Coding Region. AB - Background: The microsporidian Septata intestinalis, recently suggested to be reclassified as Encephalitozoon intestinalis, is probably the second most common microsporidian isolated from AIDS patients after Enterocytozoon bieneusi. S. intestinalis causes a disseminated disease, including infections of the gastointestinal tract, whereas E. bieneusi is confined strictly to the gastrointestinal tract. It is important to differentiate between these two microsporidians, as only infections caused by S. intestinalis can, at this time, be effectively treated. Currently, diagnosis of infections caused by S. intestinalis can be achieved only by transmission electron microscopy. Methods and Results: In this study are described specific polymerase chain reaction primers for diagnosis of S. intestinalis infections based on the region coding for the small subunit ribosomal RNA cloned from a S. intestinalis isolate. These primers were tested for specificity on cloned ribosomal RNA sequences of different species of microsporidia, as well as on cultured samples of E. bieneusi, Encephalitozoon cuniculi, Encephalitozoon hellem and Vittaforma corneae (Nosema corneum), without showing any cross-amplification. By use of these polymerase chain reaction primers, eight different microsporidian isolates grown in culture and one diagnostic sample, collected as an ethanol-fixed duodenal jejunal segment, were identified as S. intestinalis. Conclusion: These primers are powerful diagnostic tools and can enhance or replace traditional methods used to diagnose this microsporidian. PMID- 10462590 TI - Evaluation of Two Methods for Quantitation of Hepatitis C Virus RNA. AB - Background: Accurate quantitation of hepatitis C virus (HCV) RNA in serum may provide a means to predict disease course and response to interferon-alpha therapy. Several quantitative assays are commercially available, but none have been accepted as the gold standard. Methods and Results: The branched DNA quantitative hybridization assay (Quantiplex HCV 1.0, Chiron, Emeryville, CA) and a quantitative reverse transcription polymerase chain reaction (Amplicor HCV MONITOR, Roche Diagnostic Systems, Branchburg, NJ) were compared using a panel of 53 sera from patients with chronic hepatitis C. All sera contained HCV RNA of known genotype. Overall, there was a positive correlation between the results for the 41 sera that gave discrete values in both tests (r =.81, linear regression; P <.01, Kendall's rank test); however, the mean number of HCV copies per milliliter was 13.5-fold higher with Quantiplex (P <.01). A plot of the difference between methods against their means showed poor agreement between the methods. No correlation between the results of the two tests was observed for sera with MONITOR values greater than 5.0 x 10(5) copies/mL. Discrete MONITOR values were obtained for all 12 sera that were below the lower limit of quantitation of Quantiplex (mean, 1.78 x 10(5)). Parallel testing of serial dilutions of two sera showed that each method gave linear responses over the stated dynamic ranges; however, the proportional systematic error was greater with MONITOR. The mean coefficient of variation for replicate determinations was 23% for Quantiplex and 45% for MONITOR (P =.13). Conclusions: Despite a positive correlation, systematic differences exist between the two methods for quantitation of HCV RNA and they cannot be used interchangeably. PMID- 10462592 TI - Modified Method for the Detection of the CAG Repeat Expansion in Huntington's Disease and Application to a Predictive Testing Protocol. AB - Background: The identification of the CAG trinucleotide repeat expansion as the cause of Huntington's disease (HD) has dramatically altered the ease and uptake of testing. The direct test for the mutation allows testing of many more consultands, particularly those individuals whose family structure is not suitable for linkage analysis. Therefore, protocols that can rapidly handle a number of samples and give accurate reliable results are essential. Methods and Results: The HD1/HD2 set of primers, which amplify the variable CAG and polymorphic CCG repeats, and the HD1/HD3 set of primers, which amplify only the variable CAG repeat, were used. Comparison of internally labeled with end-labeled polymerase chain reaction product was made. "Lysates" made from blood were investigated as suitable material for the HD polymerase chain reaction. Conclusions: The conditions used for detection of the CAG repeat in the huntingtin gene by end labeling of one of the primers that amplifies only the CAG repeat were improved, and an efficient protocol that reduces sample preparation and storage by using lysates from blood rather than extracted purified genomic DNA was developed. PMID- 10462593 TI - Risk of Venous Thrombosis in Carriers of a Common Mutation in the Homocysteine Regulatory Enzyme Methylenetetrahydrofolate Reductase. AB - Background: Elevated levels of homocysteine are an independent risk factor for venous thrombosis. A common mutation in methylenetetrahydrofolate reductase (MTHFR), an enzyme required for efficient homocysteine metabolism, creates a thermolabile (tl-) enzyme with reduced activity that may predispose to hyperhomocysteinemia. Methods and Results: To assess whether this common mutation is a risk factor venous thromboembolism, a polymerase chain reaction-based genotyping assay was used to compare the prevalence of this mutation in a group with thrombosis versus several control groups. Of the 331 thrombosis subjects, 47% were heterozygous and 11% homozygous for tl-MTHFR. In comparison, heterozygotes constituted 42-47% and homozygous 15-16% of each of three control groups (totaling 593 subjects). There was no significant difference in the tl MTHFR homozygote frequency or allele frequency between the thrombosis and control study groups. Although the prevalence of the factor V R506Q (Leiden) mutation causing activated protein C resistance was significantly higher in the thrombosis (19%) than in the control groups (4-9%), the concomitant presence of tl-MTHFR with factor V R506Q did not contribute to any excess thrombotic risk. Conclusions: Although the tl-MTHFR mutation may predispose to hyperhomocysteinemia, a known risk factor for venous thrombosis, this common genotype is not a direct genetic risk factor for venous thrombosis, either alone or in combination with the factor V R506Q mutation. PMID- 10462595 TI - Clinical News Update. PMID- 10462594 TI - Molecular Assessment of Clonality in Lymphoproliferative Disorders: II. T-cell Receptor Gene Rearrangements. AB - T-lineage non-Hodgkin's lymphomas are a diverse group of malignancies for which routine histology and immunophenotyping alone may not be diagnostic. Molecular genetic studies can be used to detect unique T-cell receptor gene rearrangements and establish clonality in these cases. The methods for evaluating T-cell receptor clonality are less well studied and produce more varied results than those for the immunoglobulin genes. Whereas clonality is a hallmark of malignancy, its presence or absence in T-lineage disorders does not always correlate with subsequent biologic behavior. Since the molecular testing of T lineage lymphoproliferative disorders is increasing, this review summarizes the polymerase chain reaction and Southern blotting methods for T-cell receptor gene rearrangements. Interpretation of the results of these methods should always be done in the context of complete clinical and histologic data. PMID- 10462596 TI - Evaluating Children in the Ukraine for Colonization With the Intestinal Bacterium Oxalobacter formigenes, Using a Polymerase Chain Reaction-based Detection System. AB - Background: Oxalobacter formigenes is a recently discovered anaerobic bacterium residing in the gastrointestinal tracts of most vertebrates, including humans. Evidence suggests that this bacterium plays an important symbiotic relationship with its hosts by regulating oxalic acid homeostasis. Oxalic acid is a ubiquitous toxic by-product of metabolism associated with numerous pathologic conditions, including hyperoxaluia, cardiac myopathy and conductance disorders, kidney stones, and even death. Despite the potential importance of O. formigenes in several major health disorders, the difficulty in culturing, isolating, and identifying this fastidious anaerobe has limited research of its disease associations. Because O. formigenes must use two unique enzymes to catabolize oxalic acid, this bacterium appeared to be a suitable model for DNA-based identification, thereby circumventing the labor-intensive procedures currently used. Methods and Results: In this study, genus- and group-specific oligonucleotide sequences were designed corresponding to homologous regions residing in the oxc gene that enodes for oxalyl-coenzyme A decarboxylase. A polymerase chain reaction (PCR)-based amplification of the 5'end of this gene directly from genomic DNA isolated from various strains of O. formigenes was used to show that the genus- and group-specific oligonucleotide probes could identify and subgroup the bacterium. Field testing of this PCR-based detection system with 100 fecal cultures collected from children aged 0-12 years demonstrated the ease and efficacy with which O. formigenes can now be identified. Furthermore, these latter data provide a profile for the natural colonization of a human population with this intestinal bacterium. Conclusions: Development and use of this PCR based detection system permit the rapid identification and classification of the gut-associated bacterium O. formigenes, thereby circumventing the need for the more labor-intensive and lengthy method currently used. The first field test of this detection system indicates that humans apparently do not become colonized with O. formigenes until they begin crawling about in the environment. Furthermore, studies investigating the association between several disorders (eg, kidney stones, irritable bowel syndrome, and hyperoxaluria) and the absence of the bacterium from the gut will now prove far easier. PMID- 10462597 TI - Sequence-specific Priming and Exonuclease-released Fluorescence Assay for Rapid and Reliable HLA-A Molecular Typing. AB - Background: The rapid and sensitive method described for low-resolution DNA typing of alleles at the HLA-A locus is based on a sequence-specific primer polymerase chain reaction approach that eliminates post-polymerase chain reaction gel electrophoresis to analyze the results. Methods and Results: This method takes advantage of the 5' --> 3' exonuclease activity of the Taq polymerase normally present during polymerase chain reaction. This sequence-specific priming and exonuclease-released fluorescence assay was conducted on 40 DNA samples derived from homozygous cell lines and peripheral blood lymphocytes that represented the majority of the HLA-A alleles. There was 100% condordance with the results obtained by serologic typing of these same samples. Conclusions: The advantages of this system are that assays can be performed in 2 hours, the results are easy to interpret, and no gel electrophoresis is required. The same strategy can be used for high-resolution DNA typing once appropriate primer pairs are added to the panel used for first-level discrimination. PMID- 10462598 TI - Novel Chemiluminescent Substrate and Probe Systems for the Identification of CFDeltaF508 Genotypes. AB - Background: Chemiluminescence detection systems are rapidly gaining popularity as safer alternatives to isotopic methods in molecular diagnostics with equal sensitivity and specificity. In addition, they offer versatility of detection because of the availability of different haptens for labeling the probes, the antihapten antibodies conjugated with either alkaline phosphatase (AP) or horseradish peroxidase (HRP), and their respective chemiluminescent substrates. A novel dual chemiluminescent substrate (AP and HRP based) and probe systems to distinguish genotypes of cystic fibrosis DeltaF(508) mutation are described. Methods and Results: Two methodologies have been formulated to identify positively the genotypes of the cystic fibrosis DeltaF(508) mutation. In method 1, a pair of oligonucleotides designed to anneal to the fanking regions of DeltaF(508) mutation are differentially labeled with the hapten biotin or fluorescein and ligated using the template DNA of wild-type (N/N), heterozygous (N/DeltaF(508)), and homozygous (DeltaF(508)/DeltaF(508)) genotypes. The ligated product containing both labels is detected by first binding with avidin-HRP and anti-fluorescein-AP followed by reaction with the dual substrate. As expected, the ligation products are detected only in n/DeltaF(508) and DeltaF(508)/DeltaF(508) genotypes but not in N/N, where the ligation is precluded by the presence of three intervening nucleotides. In method 2, the three genotypes are hybridized on a membrane simultaneously with uniquely labeled (biotin or digoxigenin) oligonucleotides each designed to bind either the normal or the mutant allele. On treatment with HRP- and AP-conjugated antibodies followed by reaction with the dual substrate, only the band from N/DeltaF(508) genotype emitted a strong signal because of the binding of both oligonucleotides. Conclusions: The ligation and hybridization methods in conjunction with the dual substrate can detect and differentiate the genotypes with the DeltaF(508) mutation. These formats may be valuable for distinguishing normal individuals from carriers in population screening and fetuses that are heterozygous, from those that are homozygous for cystic fibrosis DeltaF(508) in prenatal and neonatal diagnosis. PMID- 10462599 TI - A Competitive Allele-specific Oligomers Polymerase Chain Reaction Assay for the cis Double Mutation in AMPD1 That Is the Major Cause of Myo-adenylate Deaminase Deficiency. AB - Background: Myo-adenylate deaminase deficiency (mADD) is the most common enzyme deficiency restricted to skeletal muscle, with a frequency of 1-2% in frozen muscle biopsies and complaints of easy fatigue and muscle cramping on exertion. A double C > T transition at coding bases 34 in exon 2 and 143 in exon 3 is the main cause of mADD. A 1-day assay using allele-specific oligomers and no isotope would be valuable for single cases. Methods and Results: Downstream primers with penultimate mismatch and 3' terminus matching the mutant or the normal base in exons 2 and 3 are used with a common upstream primer for each exon, to give amplimers of 150 bp for exon 2 and 200 bp for exon 3. A short common primer further downstream in exon 3 provides a competing 300-bp apmlimer whose product contribution is readily controlled by adjusting the annealing temperature. The entire procdure could be done in 1 day: DNA isolation, polymerase chain reaction (PCR), electrophoresis in agarose gel with ethidium bromide, and visualization by ultraviolet light. Deficient individuals have bands only with the mutant primers, normal persons have bands only with the normal primers, and heterozygotes (carriers) show bands with both primer sets. The empty slots show the 300-bp competing band, proving the PCR amplified the correct template. Allele-specific oligomers PCR results were verfied by dot blots and by restriction endonuclease analysis of exon 2. Conclusions: A simple and reliable allele-specific PCR assay using DNA from blood (or muscle) is now available for the diagnosis of individual cases of mADD caused by the common double-mtant AMPD1 gene, including the rare instances arising from homologous recombination between the two mutations. PMID- 10462600 TI - Two Novel Mutations in the Cystathionine beta-synthase Gene of Homocystinuric Patients. AB - Background: The continued identfication of new mutations in the cystathionine beta-synthase (CBS) gene is important in correlating the genotype/phenotype of patients with classic homocystinuria and in assessing whether heterozygosity of CBS deficiency is an important cause of mild hyperhomocysteinemia, an independent risk factor for occlusive vascular diseases. Methods and Results: Single-strand polymorphism and direct nucleotide sequencing were used to detect two novel mutations in the CBS gene of three homocystinuric patients from two unrelated families. The first mutation, a G-to-A transistion at nucleotide 1316 in exon 12, results in an amino acid substitution of arginine by glutamine at codon 439. The second mutation is a G-to-A transition at nucleotide 1109 in exon 10 and results in an amino acid substitution of cysteine by tyrosine at codon 370. All three patients are apparently compound heterozygotes, with one of the two novel mutations on one allele and the T(833)C mutation on the other allele. Conclusions: The absence of the G(1316)A and G(1109)A in 216 control alleles demonstrates that these two novel mutations do not represent common polymorphisms, but rather are responsible for the defective CBS enzyme activities encoded by one of the two alleles of the CBS gene in each of the two families. PMID- 10462601 TI - Enhanced Reverse Transcriptase Polymerase Chain Reaction for Prostate-specific Antigen Combined With Needle Biopsy Results: A Superior Predictor of pT3 Disease. AB - Background: Preoperative staging for prostate cancer underestimates the final pathology stage in approximately 40-50% of the cases. Previous work from our institution demonstrated that an enhanced reverse transcriptase polymerase chain reaction (RT-PCR) assay for prostate-specific antigen (PSA) enabled more accurate staging of presumably localized prostate cancer. The goal of the current study is to determine if needle biopsy results when combined with the RT-PCR for PSA assay are a better predictor of final pathology stage. Methods and Results: One hundred sixty-two men with needle biopsy-diagnosed prostate cancer had blood drawn for the RT-PCR for PSA assay before undergoing radical prostatectomy. Polymerase chain reaction primers specific for the PSA gene were run, along with appropriate controls. Tumor was characterized using the TMN staging system: organ confined (pT2a-c), capsular penetration (pT2a-b), seminal vesicle involvement (pT3c). Surgical margins and lymph nodes were also evaluated. Of the 162 patients, the majority had localized disease by digital rectal examination: T2 = 97%, and T3 = 3%. On needle biopsy, 48 cases (30%) had a Gleason score >/=7 and 35 cases (22%) had perineural involvement (PNI). The RT-PCR for PSA assay was positive in 50 patients (31%). Final pathology revealed 39% of patients had pT3 disease; none of the 162 patients had lymph node involvement. Statistical analysis revealed that a Gleason score >/=7 had 81% specificity and 46% sensitivity in predicting pT3 disease (odds ratio 3.6). The presence of PNI on needle biopsy was 89% specific and 38% sensitive in predicting pT3 disease (odds ratio, 4.9). The RT-PCR for PSA assay was 89% specific and 62% sensitive in predicting pT3 disease (odds ratio, 13.0). All 14 cases with both RT-PCR for PSA and PNI positivity had pT3 disease. Logistic regression analysis demonstrated the independent predictive strength of PNI on needle biopsy, Gleason score >/=7, and RT-PCR for PSA positivity for identifying pT3 disease; their combined odds ratio was more than 180. Conclusions: Using the RT-PCR for PSA assay in conjunction with needle biopsy results increases the predictive strength for pT3 disease in patients with presumed organ-confined prostate carcinoma. PMID- 10462603 TI - Clinical News Update. PMID- 10462602 TI - Gene Therapy Monitoring: Clinical Monitoring for Efficacy and Potential Toxicity. AB - Since the first human clinical trial was initiated in 1990, the field of gene therapy has gone through a period explosive growth. Building on a sound foundation of many years of basic and clinical research this novel therapeutic approach has the potential to revolutionize medicine. As gene therapy moves from a research setting, however, clinical monitoring for efficacy and potential toxicity will become increasingly important, perhaps becoming as commonplace as therapeutic drug monitoring is today. PMID- 10462604 TI - Clinical News Update. PMID- 10462605 TI - Detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Ureaplasma urealyticum, and Mycoplasma genitalium in First-void Urine Specimens by Multiplex Polymerase Chain Reaction. AB - Background: Sexually transmitted diseases are often caused by one or more microorganisms, and asymptomatic carriage and transmission may be of significance. Testing for more than one organism in a single assay could be a useful approach to laboratory diagnosis. Methods and Results: A multiplex polymerase chain reaction (PCR) assay was developed that employed specific primers targeted to the 7.5-kb cryptic plasmid of Chlamydia trachomatis, the cppB gene of the 4.2-kb cryptic plasmid of Neisseria gonorrhoeae, the 140-kd major adhesion protein gene of Mycoplasma genitlium, and the urease gene of Ureaplasma urealyticum. All four polymerase chain reaction products were detectable by agarose gel electorphoresis and were confirmed by Southern hybridization using fluorescein isothiocyanate-labeled oligonucleotide probes and enhanced chemiluminescent detection. Using purified DNA preparations, multiplex PCR had a reproducible detection limit of 1 fg of C. trachomatis DNA, 100 fg of N. gonorrhoeae DNA, and 10 fg U. urealyticum DNA and M. genitalium DNA, which converts to 1-2 genomic equivalents (ge) of C. trachomatis and N. gonorrhoeae, 4 ge of M. genitalium, and 10 ge U. urealyticum. Multiplex PCR was compared with individual uniplex polymerase chian reaction PCR assays by testing 117 first-void urine samples (91 men, 26 women) from Canadian or Kenyan patients. Multiplex PCR detected 45 of 46 (97.8%) urines with C. trachomatis DNA, 42 of 42 (100%) urines with N. gonorrhoeae DNA, 17 of 17 (100%) urines with U. urealyticum DNA, 4 of 4 (100%) urines with M. genitalium DNA, 12 of 12 urines that had DNA from two bacteria, and 2 of 2 urines with DNA from three bacteria. Multiplex PCR correctly identified bacteria in 92 of 93 urines for an overall sensitivity of 98.9%. Specificity calculations were 100% for C. trachomatis (71/71), N. gonorhoeae (75/75), U. urealyticum (100/100), and M. genitalium (113/113). Conclusions: Multiplex PCR provided a single sensitive and specific test for the detection of four bacteria in first-void urine samples. Testing of first-void urine samples by multiplex PCR could facilitate studies aimed at improving our understanding of the epidemiology of these important sexually transmitted diseases. PMID- 10462606 TI - Simplified Reverse Dot Blot Analyses for Detecting of ras Oncogene Mutations. AB - Background: Mutations in members of the ras gene family (H-ras, K-ras, and N-ras) have been identified in various human malignancies. A variety of techniques have been used to test for ras mutations. Methods and Results: A simplified reverse dot blot (RDB) assay was used in this study. Polymerase chain reaction products were hybridized to nitrocellulose membrane-fixed synthetic probes (20 nucleotides long) specific for codons 12, 13, and 61 of H-, K-, and N-ras mutations and their wild-type sequences. No special treatment or modification of the probes was necessary to obtain adequate results in overnight film exposure when the polymerase chain reaction was carried out using (32)P-end labeled primers. It was demonstrated that this simplified RDB assay can also be used with fluorescein-11 dUTP and a chemiluminescence detection system. The RDB assay is more reliable than the single-strand conformation polymorphism (SSCP) assay. By comparison, the SSCP assay is significantly less sensitive and less specific. It was confirmed with sequencing that 11 (12%) of 93 SSCP assays were false positive and 2 (2%) were false negative, whereas no false positive or false negative RDB assay was detected. The RDB assay also provides more additional detailed information about the specific point mutation and amino acid change, which may have clinical implications in some tumors. Conclusions: The RDB assay is very sensitive and able to detect mutations when the mutant allele is in 1% of the cells and can be used to detect minimal residual disease, particularly in some cases of leukemia and myelodysplasia. PMID- 10462607 TI - Use of Molecular Diagnostic Methods for Lymphoma Staging in Bilateral Bone Marrow Biopsies. AB - Background: Testing for immunoglobulin and T-cell-receptor gene rearrangement is a useful adjunct to morphologic evaluation of bone marrow involvement by non Hodgkin's lymphoma. At this institution, each sample from a patient undergoing bilateral bone marrow sampling for lymphoma staging is assayed seperately to avoid possible false negatives from dilution, if, for example, only one sample is positive. Methods and Results: Gene rearrangement results were reviewed for 140 patients, to assess if pooling the two aspirate samples from bilateral biopsies could be implemented. Discordant results between right and left iliac crest samples were found in 4 of 140 (2.9%) patients. Morphologic assessment was performed for 138 of the 140 patients. In 64 cases there were no abnormal findings. Four of these 64 (6.3%) had monoclonality demonstrated by genotyping or immunophenotyping. Of 134 patients for which both tests were performed, 17 (12.7%) showed discordance: 6 were negative by immunophenotyping, positive by genotyping (3 cases with malignant morphology, 1 with suspicious morphology, and 2 without evidence of malignancy); 11 positive by phenotyping but not genotyping (6 cases with malignant morphology, 3 with suspicious morphology, and 2 without evidence of milignancy). Conclusions: These findings support the role for these ancillary studies; in some cases both may be needed to demonstrate monoclonality. Based on this low occurence of discordant findings between right and left samples, we feel that bilateral specimens can be pooled without unacceptable risk of false negatives due to possible dilution. PMID- 10462608 TI - Mycobacterium tuberculosis DNA Not Detected by Polymerase Chain Reaction In Air Samples From Hospital Rooms of Tuberculosis Patients. AB - Background: As the incidence of tuberculosis (TB) increased in the United States, the risk of occupational and nosocomial TB also increased. Airborne Mycobacterium tuberculosis (MTB) is difficult to measure directly. Quantifying MTB DNA by polymerase chain reaction in air samples obtained from isolation rooms of patients with TB would provide a measure of the number of airborne organisms produced by a patient and the efficacy of ventilation, and might predict when an individual patient is no longer infectious. Methods and Results: The air was sampled through cellulose ester filters from the isolation room of a patient with newly diagnosed pulmonary TB, from a patient on therapy for 14 days, and from adjacent offices, and an attempt was made to detect MTB DNA; however, MTB DNA was detected only on positive control filters. Conclusions: Mycobacterium tuberculosis was not detected by polymerase chain reaction in air samples from the rooms of two patients with pulmonary TB. This may have been due to the large number of room air changes with resultant rapid clearance of airborne droplet nuclei or to the limited air volume sampled. A sensitive molecular assay of airborne MTB could be used to monitor the efficacy of infection control measures by sampling a sufficient volume of isolation room air and could aid in determining when an individual patient was no longer infectious. PMID- 10462609 TI - Temporal and Quantitative Analysis of Chimerism in Liver and Kidney Transplant Patients: An Enhanced Fluorescence Detection System Allows for More Sensitive Quantitative Detection. AB - Background: Because spontaneous microchimerism has been reported in stable renal and hepatic allografts, the presence of donor-derived cells in recipient tissues was investigated in kidney and liver tranplant recipients. Methods and Results: Human lymphocyte antigen class II markers and Y-chromosome sequences in male donor-to-female recipient transplants were used for chimeric analysis. Human lymphocyte antigen typing was performed by group-specific polymerase chain reaction amplification and restriction fragment length polymorphism analysis, X chromosome- and Y-chromosome-specific primers were used in a multiplex polymerase chain reaction analysis. Quantitative Y-chromosome analysis was performed using energy-transfer fluorescence from a nested primer system. Patients who had rejected their grafts were also analyzed, as were a group who were analyzed for chimerism at the time of transplant (day 1) and sequentially at various intervals for up to 3 months. Of 23 long-term kidney patients analyzed, 16 were chimeric by human lymphocyte antigen or sex-determination analysis. In 2 patients whose graft had failed no chimerism was observed. Chimerism in liver patients was detectable on the day of transplant and was maintained for 30 to 120 days as measured at 5 day intervals (these patients continue to be monitored). Quantititative analysis suggested that the ratio of donor to recipient cells was variable in a patient and ranged from greater than 1 in 10(4) to less than 1 in 10(5). An enhanced fluorescence energy-transfer detection system was adopted to increase sensitivity of the polymerase chain reaction detection of chimerism and to quantitate the results. Conclusions: The results indicate that cells from the donor organ migrate into recipient tissues early after transplantation. These cells persist in a majority of patients for at least 3 to 4 years. It has been proposed that tolerance is related to the presence of these "passenger" leukocytes and that dendritic cells play the most important role. The data suggest that the establishment of chimerism plays an important role in graft acceptance in a majoritiy of the kidney and liver patients in this study. These findings also suggest that the levels of chimeric cells, "a quantitative chimerism," may be important in establishing tolerance but further studies are required to support this contention. PMID- 10462610 TI - CD44 Expression in Benign and Neoplastic Human Prostates. AB - Background: CD44, a major cell surface receptor for hyaluronic acid, is a family of ubiquitous cell surface glycoproteins. Altered levels of CD44 expression, seen in many epithelial neoplasms, have prognostic implications. Expression of standard and variant isoforms of CD44 was assessed in normal and neoplastic human prostate tissue and culture cells to evaluate as a marker for malignant transformation. Methods and Results: Expression of CD44s, CD44R, v5, v6, v7/8 and v10 was assessed in prostate tissue (benign and malignant) and cell lines (DU 145, PC-3, LNCaP, p69) and primary cultures of normal prostates and adenocarcinoma cells obtained from prostatectomies using reverse transcriptor polymerase chain reaction, Western blotting, and immunofluorescence. No CD44 expression was seen in LNCaP cells. p69, DU-145, and PC-3 cells expressed CD44s and CD44R. p69, cells demonstrated a 1000-bp-long form of CD44 mRNA, unique to this normal cell line. Both normal and neoplastic prostatic tissue demonstrated CD44s on Western blotting. Conclusions: In agreement with previous studies, prostatic adenocarcinoma cells, except LNCaP, expressed CD44s. Different patterns of CD44 expression were seen in benign and neoplastic prostate. Benign prostate exhibited higher v5 protein levels, whereas neoplastic prostates demonstrated higher CD44s expression. CD44s expression was identified in all neoplastic prostates as compared with only 50% of the benign prostates. No significant difference in expression of the other variants assessed (v6, v7, v7/8, and v10) was observed in the benign and neoplastic prostates. PMID- 10462611 TI - A Novel Missense Mutation, R1283S, of the Cystic Fibrosis Transmembrane Conductance Regulator Gene in a 47-Year-Old African-American Patient. AB - Background: More than 600 mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene have been described; however, at least 50% of the disease-associated mutations in the African-American population remain unknown. Reported here is a novel missense mutation, R1283S, in a 47-year-old African-American patient with mild cystic fibrosis. Methods and Results: The patient was screened for 27 common and less common CFTR mutations and 2 mutations were detected. Direct sequencing confirmed the presence of the DeltaF508 mutation and revealed the presence of a novel missense mutation, R1283S. Conclusions: R1283S appears to be a cystic fibrosis mutation associated with mild disease, and adds to the number of known mutations in African-Americans. R1283S can be confused with the more common mutation, W1282X, when polymerase chain reaction restriction fragment length polymorphism analysis is used for detection. PMID- 10462612 TI - Evidence-based Health Care. PMID- 10462614 TI - Clinical News Update. PMID- 10462613 TI - Optimization of the Isolation and Amplification of RNA From Formalin-fixed, Paraffin-embedded Tissue: The Armed Forces Institute of Pathology Experience and Literature Review. AB - Background: RNA is extensively degraded by routine formalin fixation to fragments averaging 200 nucleotides (nt). Several methods for the recovery of amplifiable RNA from formalin-fixed, paraffin-embedded tissue have been described; however, a universally accepted approach in a clinical molecular diagnostic laboratory has not yet emerged. Methods and Results: Amplifiable RNA can be recovered with high efficiency from all types of formalin-fixed, paraffin-embedded tissue using proteinase K digestion, either a phenol-chloroform or an acidic guanidinium thiocyanate-phenol chloroform extraction step, and isopropanol precipitation in the presence of glycogen. Designing primers to detect a small target was critical for consistent RNA amplification in the following assays, with the target size indicated: hepatitis C virus, 169nt; morbillivirus, 78 nt; influenza virus, 113 nt; the npm-alk fusion product resulting from t(2;5) translocation, 175 nt; and the bcr-abl fusion product resulting from t(9;22) translocation, 93 or 168 nt. Conclusions: With use of beta-2-microglobulin as the control messenger RNA target for assessing the recovery of amplifiable RNA from human tissue, amplifiable RNA was recovered from 216 of 225 blocks (96%). In a series of veterinary specimens, which were largely postmortem and moderately to severely autolyzed, 158 of 199 blocks (79%) yielded amplifiable RNA using a beta-actin target. Amplifiable influenza RNA has been recovered from archival paraffin blocks as old as 79 years. PMID- 10462615 TI - Modified Method for Competitive Reverse Transcription Polymerase Chain Reaction for Rapid and Automated Quantitation of Messenger RNA in Multiple Samples. AB - Background: Competitive reverse transcription polymerase chain reaction (RT-PCR) has been used increasingly to quantitate messenger RNA (mRNA) levels; however, conventional competitive RT-PCR methods require four or five reactions per sample of RNA, employing serial dilutions of an internal competitor sequence, making analysis of multiple samples of tedious process. A modified method is described by which multiple samples and multiple RNA transcripts can be analyzed easily by an automated process. Methods and Results: Transforming growth factor beta-1 (TGF beta-1) mRNA was assayed in total RNA extracted from cultured human skin fibroblasts. A standard solution of total RNA was first prepared by pooling RNA from several cell lines and stored in aliquots. A 270-bp competitor RNA molecule (RNA mimic) was prepared by in vitro transcription and was added to each reaction. PCR was performed with a fluorescent dye (Hex; Applied Biosystems, Foster City, CA)-labeled sense primer to amplify a 161-bp-long c DNA product of target TGF-beta-1 mRNA sequence and the RNA mimic. The PCR products were analyzed with an automated laser-scanned gel electrophoresis system and the area under the curve (AUC) was used for quantitation. The concentration TGF-beta-1 mRNA in standard RNA was determined by conventional competitive RT-PCR. Subsequently, equal amounts of RNA mimic were mixed with four serial dilutions of standard RNA and 0.1 ug of sample total RNA for RT-PCR. A standard curve was generated using the known dilutions of a standard target RNA solution and ratio of AUC for target to that for mimic for each dilution. The unknown sample was then quantitated by interpolation of its area under the curve ratio on the standard curve. This method had inter- and intra-assay coefficients of variation of less than 10%. Conclusions: This modification is highly reproducible for quantitation of mRNA and significantly reduces the number of PCR reactions required for each assay. It can be used to assay several RNA molecules in a given sample by designing RNA mimics and PCR primers to generate PCR products of different lengths so that they can be analyzed by the laser scanning of a single lane of electrophoretic gel. PMID- 10462616 TI - Spinal Muscular Atrophies: An Ongoing Diagnostic Dilemma? AB - Background: Spinal muscular atrophies (SMAs) are a group of autosomal recessive disorders of anterior horn cell degeneration. Three genes-survival motor neuron (SMN), neuronal apoptosis inhibitory protein (NAIP), and, more recently, p44 (subunit of basal transcription factor II)-have been considered as candidate genes. The region spanning these genes has a complex organization, which makes molecular analysis difficult. Methods and Results: Molecular genetic testing of deletions of exons 7 and 8 of the SMN(T) (telomeric copy) gene and exon 5 of the NAIP(T) (telomeric copy) gene was performed in 39 diagnosed SMA patients, 31 cases referred as possible SMA, and 24 cases of prenatal diagnosis of SMA. Linkage analysis using markers flanking the SMA region was also performed. In general, the findings of involvement of SMN and NAIP gene deletions in patients diagnosed with SMA are in agreement with those previously published. One possible SMA case was found to be homozygously deleted only for exon 7 of SMN(T) and one deleted only for exon 5 of the NAIP(T) gene. Conclusions: SMAs exemplify human inherited disorders in which application of a variety of different techniques and a search for mutations in multiple genes are involved. Deletion testing of candidate genes (SMN, NAIP) is a powerful approach in patients affected or suspected of being affected with SMA. It is proposed that the direct SMN gene deletion test can be offered as the only test for prenatal diagnosis of SMA in families in which the clinically affected sibling has also been shown to have the homozygous deletion. PMID- 10462617 TI - Identification of Deletion Carriers in Duchenne/Becker Muscular Dystrophy Families Using a Digoxigenin-Labeled Quantitative Polymerase Chain Reaction Technique. AB - Background: A quantitative polymerase chain reaction (PCR) technique based on the incorporation of digoxigenin (DIG), and visualization of the labeled fragments for the detection of deletion carriers in Duchenne/Becker muscular dystrophy families has been developed. Methods and Results: Sixty-five DNA samples taken from mothers and/or sisters of familial and sporadic deletion patients were investigated in the exponential phase of amplification. All obligate carriers were correctly identified using this technique. In more than 95% of deletion families, possible carriers could be screened by using four different multiplex systems specifically designed to increase the efficiency of the detection. Deletions were found to be present in 42% of possible carriers. All these results were confirmed by computer-assisted laser densitometry. Conclusions: Dosage analysis by DIG-labeled quantitative PCR is a reliable and accurate technique for detecting Duchenne/Becker muscular dystrophy deletion carriers. PMID- 10462618 TI - Automated Detection of Trinucleotide Repeats in Fragile X Syndrome. AB - Background: The conventional method for diagnosis of fragile X syndrome has been amplification of the trinucleotide repeat region of the FMR-1 gene by polymerase chain reaction (PCR) and Southern blot analysis to detect full expansion and hypermethylation. "Stuttering" resulting from incomplete amplification is still observed in the PCR products despite the use of reagents that reduce the secondary structure of the GC-rich template. In addition, PCR products can be detected by autoradiography only after 1 to 2 days of exposure. By combination of a recently reported amplification protocol with fluorescence detection of PCR products in an automated DNA sequencer, the PCR protocol for amplification of trinucleotide repeats was simplified. This modified protocol is highly reproducible, more accurate, and less costly than the conventional protocol because of the elimination of radioisotopes from the PCR. Methods and Results: PCRs were conducted with betaine and Pfu DNA polymerase. This improved PCR protocol allowed immediate detection of PCR products in agarose gels containing ethidium bromide. Stuttering was completely eliminated and fragments of up to 1kb ( approximately 250 repeats) were visible in agarose gels. PCR products were automatically detected by laser fluorescence in an automated DNA sequencer by inclusion of a fluorescently-labeled primer in the PCR reaction. A short electrophoresis run of 100 minutes in denaturing acrylamide gels was sufficient to give high resolution of fragments with higher accuracy and sensitivity than conventional detection by autoradiography. Conclusions: A simple, nonradioactive protocol that is more rapid and less expensive than the conventional PCR protocol for the detection of trinucleotide repeats has been developed. By use of this detection protocol, fragment sizes containing up to 100 repeats could be detected, alleles differing by one trinucleotide repeat were clearly resolved, and heterogeneous repeat patterns such as those present in mosaics could be discriminated. This protocol has been adapted to the amplification and detection of at least two other classes of trinucleotide repeats [(CAG)(n) and (CTG)(n)], suggesting that it may be a universal protocol for PCR amplification and detection of trinucleotide repeats. PMID- 10462619 TI - Optimization of Apolipoprotein E Genotyping. AB - Background: Three common alleles of apolipoprotein E (apoE) have been identified and are expressed codominantly to generate six genotypes. Different apoE genotypes are implicated in several cardiovascular and neurologic disorders. Testing for apoE genotypes has increasing diagnostic importance, particularly in the risk assessment of coronary artery disease. A reproducible and cost-effective assay was developed. Methods and Results: Polymerase chain reaction (PCR) amplification of the fourth exon of the apoE gene is performed in the presence of dimethyl sulfoxide using two-step thermal cycling. The PCR products are digested with HhaI restriction enzyme and analyzed by agarose gel electrophoresis to determine apoE genotypes. Effects of several factors, including dimethyl sulfoxide, DNA concentration, and PCR cycling conditions, on PCR specificity and efficiency have been determined and optimized. Conclusions: Apolipoprotein E genotyping by a PCR restriction fragment length polymorphism analysis has been optimized for use in a clinical diagnostic laboratory, allowing evaluation of up to 52 samples by one technician in one day. PMID- 10462620 TI - Age-Related Disease Penetrance in a Large Medullary Thyroid Cancer Family With a Codon 609 RET Gene Mutation. AB - Background: Familial medullary thyroid cancer (MTC) is a form of type 2 multiple endocrine neoplasia in which individuals develop MTC as the sole phenotypic manifestation of their disease. A previous study has suggested that patients with familial MTC may have a later age of onset (and more indolent course) of MTC than is observed in individuals with multiple endocrine neoplasia type 2A. Methods and Results: The age-related penetrance of MRC, C-cell hyperlasia, and a positive pentagastrin test for carriers of a codon 609 mutation of the RET gene in a large MTC family was determined. Pentagastrin testing and surgical pathology findings for patients who had thyroidectomies were correlated with RET sequence analysis findings. The penetrance of this mutation for the development of MTC was 0% at age 20, 10% at age 20, 10% at age 30, 50% at age 45, and approximately 100% at age 60. The ages of onset of C-cell hyperplasia and a positive pentagastrin stimulation test were similar, and both preceded the age of onset of MTC. Carriers of the mutated gene in this family had a later age of onset of disease that has been reported for families with multiple endocrine neoplasia type 2A and 2B syndromes. Conclusions: These results may have implications for the clinical management of MTC families with a 609 mutation. PMID- 10462621 TI - Human Lymphocyte Antigen Typing: Direct DNA Typing-The Only Choice? AB - Exact human lymphocyte antigen (HLA) allele matching and sequence variation are important for matching organ donors, immune response studies, and disease association investigations. The number of HLAs has reached several hundred within each of the different classes. This level of heterogeneity makes routine DNA typing to the allele level problematic using fixed probe and primer technologies. Routine large-batch screening programs where only intermediate-level typing is required can be performed in automated fashion by several DNA technologies. Screening large numbers of samples with probe-based technologies, even oligo array chips, is cost effective. Allele-specific typing is most easily performed using direct sequencing. New sequencing technologies based on cycle sequencing and high-speed capillary gels have made routine sequencing for clinical typing a reality. The complexity of the class I locus requires a detailed analysis of all the polymorphisms within exons 2 and 3. Sequencing strategies are thus designed to use informative variable regions within the flanking introns and the flanking region as well as the untranslated regions. Simliar strategies are being adapted to the complex class II DRB alleles, which now number about 200 different alleles. Greater understanding of HLA diversity and distribution throughout humans and their relatives facilitates organ matching and the history and origins of human populations. Knowledge of parasites and their role in the selection of alleles will ultimately lead to better prediction and manipulation of the immune system response to these organisms. HLA typing is used to determine relative risk to a variety of autoimmune diseases. Future uses of molecular HLA typing may include the prevention and cessation of these self-destructing diseases. PMID- 10462622 TI - Disease Gene Patenting Is a Bad Innovation. PMID- 10462623 TI - The issues change. It's the nature of science. PMID- 10462624 TI - Multidrug resistance in human tumors--molecular diagnosis and clinical significance. AB - BACKGROUND: Multidrug resistance (MDR) of human tumors is one of the major reasons for the failure of chemotherapy in refractory cancer patients. MDR can be intrinsic or acquired, depending on the time of its occurrence, either at diagnosis or during chemotherapy. Molecular investigations in MDR during the last two decades have resulted in the isolation and characterization of genes coding for P-glycoprotein, multidrug resistance-associated protein, lung resistance related protein, drug resistance-associated protein, breast cancer resistance protein, and adenosine triphosphate-binding cassette protein. Several molecular probes, primer pairs, and monoclonal antibodies have been developed over these years to quantify the regulation and expression of these drug resistance markers in tumor cells. Methodologies have also been standardized to estimate the gene amplification, mRNA and protein expression, and functionality of drug resistance proteins in clinical specimens from cancer patients. METHODS AND RESULTS: This review describes these drug resistance genes and techniques for detection and quantification of their expression and function. CONCLUSIONS: Because these markers have clinical significance and usefulness, currently available technology warrants the application of these markers in clinical oncology. PMID- 10462625 TI - MTHFR (C677T) polymorphisms and stage III colon cancer: response to therapy. AB - BACKGROUND: Leucovorin and 5-fluorouracil (5-FU) chemotherapeutics are often used as coinhibitors of the thymidylate synthase pathway to thwart the growth of cancer cells in certain types of neoplasms. The metabolism of leucovorin is mediated through the enzyme methylenetetrahydrofolate reductase (MTHFR). A common polymorphism in the MTHFR gene has been reported to be responsible for as much as a 70% reduction in activity of this enzyme when present in the homozygous form. METHODS AND RESULTS: A total of 51 stage III colon cancer patients were identified through our tumor registry. Non-neoplastic, archived tissue was obtained for each patient and subjected to MTHFR C677T PCR-RFLP genotyping. The MTHFR C677T allele was present in 32 patients (28 heterozygotes and 4 homozygotes). The remaining 19 patients carried only the wild-type allele. Overall survival was 42.10% (8/19) for wild types and 43.757% (14/32) for those with at least one C677T allele. Of the four homozygotes identified, three have succumbed to their cancer and one is alive with cancer. CONCLUSIONS: We were unable to demonstrate a survival difference between those stage III colon cancer patients receiving leucovorin therapy that carried the MTHFR C677T allele and those that were wild type for this allele. The results of this study suggest that certain subgroups (ie, homozygotes) of patients may benefit from genotypic analysis of the MTHFR gene. PMID- 10462626 TI - An automated semiquantitative B and T cell clonality assay. AB - BACKGROUND: Rearrangements of the antigen receptor genes in B and T cells generate products of unique length and sequence. Polymerase chain reaction (PCR) assays are routinely used to identify clonal lymphocyte populations by detecting clonal V-J rearrangements or chromosomal translocations within these antigen receptor loci. Multiple primer sets are, however, required to detect the majority of clonal B- and T-cell malignancies. Products from the individual reactions must be analyzed separately to avoid misinterpretation. Moreover, small clonal populations remain difficult to identify. To address these difficulties, we propose that an integrated fluorescence-based approach to clonal B- and T-cell detection would simultaneously identify both B- and T-cell neoplasia; increase amplicon resolution, analytic sensitivity, and assay throughput; produce more comprehensive and semiquantitative data useful for evaluation of hematologic malignancies; and eliminate labor intensive agarose and polyacrylamide gel electrophoresis. METHODS AND RESULTS: Samples were genomic DNA and cDNA. Differentially labeled primers were used to amplify regions diagnostic for B- and T-cell clonality in a single plate with a single thermocycler program. Combined amplicon products underwent capillary electrophoresis for high resolution fractionation and differential fluorescence detection and quantification. Data were automatically analyzed and archived. In a comparative analysis of a variety of clinical samples, this automated and integrated B- and T-cell assay showed >94% agreement (33 of 35 results) with individual B- and T-cell PCR assays. Furthermore, this assay had an overall monoclonality detection rate of 100%, and as little as 100 ng of sample DNA yielded complete B- and T-cell clonality test results. The limit of detection was approximately 10-2 cells, and amplicons were sized to within 0.1 basepair. Serial dilutions of clonal B- and T-cell lines comprising a coded proficiency panel were identified and correctly ranked. Specificity was 100% as determined by analysis of 18 control samples that were all negative for B- and T-cell clonality. CONCLUSIONS: Our data show that this automated and integrated B- and T-cell clonality assay system is a sensitive and specific tool useful for rapid identification of clonal lymphocyte populations and will likely have broad clinical applications. PMID- 10462627 TI - Enhanced sensitivity with a novel TCRgamma PCR assay for clonality studies in 569 formalin-fixed, paraffin-embedded (FFPE) cases. AB - BACKGROUND: Clonal rearrangement of genes encoding the immunoglobulins (Ig) and T cell antigen receptors (TCR) are considered to be useful markers for the diagnosis of lymphoma and for determining the clonal origins of B- and T-cell populations in lymphoid neoplasms. METHODS AND RESULTS: Polymerase chain reaction based clonality assays for TCRgamma, TCRbeta, and immunoglobulin (Ig) heavy chain (IgH) gene rearrangements were evaluated for diagnostic sensitivity and specificity in 569 formalin-fixed, paraffin-embedded (FFPE) tissues. Combined TCRbeta and TCRgamma assays enhanced the routine detection of TCR clonality to 90% of all peripheral T-cell lymphoma (PTCL) cases. IgH clonality was detected in 59% of 241 peripheral B-cell lymphoma (BCL) cases and 6% of 169 PTCL cases. Of 452 lymphomas, 5% could not be classified phenotypically as B or T lineage after immunohistochemical and clonality studies. Of all BCL cases analyzed, 24% had detectable TCRbeta and/or TCRgamma clonality. Of these BCL with biclonal results, 47% were extranodal lymphomas from skin and various tissues. CONCLUSIONS: Clonality assays were useful for distinguishing reactive or benign lymph nodes from neoplastic lymphoid infiltrates in most cases. The inclusion of TCRb and TCRg assays in the assessment of lymphomas results in a significant increase in the sensitivity of clonality detection, but is of limited utility in assessing the T- or B-cell phenotype of the tumor. PMID- 10462628 TI - Non-PCR-dependent detection of the factor V Leiden mutation from genomic DNA using a homogeneous invader microtiter plate assay. AB - BACKGROUND: The goal of this study was to compare the Invader technology for the direct detection of the Factor V Leiden mutation from genomic DNA with a conventional polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. METHODS AND RESULTS: In the Invader assay, a specific upstream "invading" oligonucleotide and a partially overlapping downstream probe together form a specific structure when bound to complementary DNA template. This structure is recognized and cut at a specific site by the Cleavase enzyme, resulting in release of the 59 flap of the probe oligonucleotide. This fragment now serves as the "Invader" oligonucleotide with respect to synthetic secondary targets and secondary fluorescently labeled signal probes contained in the reaction mixture, resulting in specific cleavage of the secondary signal probe by the Cleavase enzyme. Fluorescence signal is generated when this secondary probe, labeled with dye molecules capable of fluorescence resonance energy transfer, is cleaved. Genomic DNAs isolated from peripheral blood buffy coats of patients previously tested for the Factor V Leiden mutation by PCR-RFLP were tested using an Invader assay specific for this mutation. In all 48 samples containing sufficient DNA for testing (30 normal, 16 heterozygous, 2 homozygous mutant), the genotype determined by the Invader assay was concordant with the PCR-RFLP results. CONCLUSION: These results indicate that a simple microtiter plate-based Invader assay can reliably genotype routine clinical patient samples for the Factor V Leiden point mutation without the need for PCR amplification, restriction enzyme digestion, or gel electrophoresis. PMID- 10462629 TI - Culture-independent prediction of isoniazid resistance in Mycobacterium tuberculosis by katG gene analysis directly from sputum samples. AB - BACKGROUND: The molecular prediction of isoniazid (INH) resistance in Mycobacterium tuberculosis is hampered by the need for specialized equipment, expertise, high costs, a limited range of detectable mutations, or several of these factors. The rationale for the study was to find a practical alternative and to demonstrate generally valid problems. METHODS AND RESULTS: DNA extracted from decontaminated sputum pellets was used to amplify a 0.26 kb target sequence within the katG gene. Mutations of codon 315, frequently found in isoniazid resistant isolates, could be discriminated in a simple agarose minigel format following an AciI digest of the nested polymerase chain reaction (PCR) product. Within a panel of 22 sputum samples, INH resistance could be predicted in 5 of 10 samples containing isoniazid-resistant M. tuberculosis. The protocol is robust, requires little expertise and no specialized equipment, and provides the test results within 2 days. CONCLUSION: The results show the feasibility to rapidly and easily detect mutations highly predictive of isoniazid resistance. Nevertheless, this, like any other molecular resistance prediction test, is affected by often neglected factors, including mutation prevalences, the phenomenon of heteroresistance, and a possible bias toward one's own method. PMID- 10462630 TI - Chromosomal aneuploidy in leukemic blast crisis: a potential source of error in interpretation of bone marrow engraftment analysis by VNTR amplification. AB - BACKGROUND: Polymerase chain reaction (PCR) amplification of polymorphic microsatellite or minisatellite DNA markers has proven to be a fast, sensitive, and specific technique in post-transplantation monitoring of bone marrow engraftment, as well as early detection of residual disease and relapse. Deletion or amplification of chromosomal segments carrying marker loci, as can occur in leukemia and other hematologic malignancies, may result in loss or increased dosage of marker alleles. Examination of these marker alleles by PCR therefore may give aberrant results, which might lead to misinterpretation of bone marrow transplantation (BMT) engraftment studies. METHODS AND RESULTS: We report a case of chronic myelogenous leukemia treated by BMT. PCR amplification of the minisatellite at the apoB locus on chromosome 2 was used to monitor the donor bone marrow engraftment. The patient experienced relapse in blast crisis with a near-haploid karyotype with loss of recipient-specific apoB allele causing an aberrant PCR result for bone marrow engraftment that mimicked full donor engraftment. CONCLUSIONS: Loss or gain of polymorphic DNA markers because of chromosomal losses or gains in some hematologic malignancies may affect the interpretation of bone marrow engraftment studies by PCR. When choosing polymorphic markers for such studies, it is important to avoid those that will be affected by expected chromosomal alteration, if possible. In addition, any abberant post-transplantation typing should prompt further investigation to rule out the possibility of chromosomal aberration. Review of all pertinent laboratory studies is important to avoid misinterpretation of results from a single test for engraftment analysis. PMID- 10462631 TI - Development of an internal restriction control in the PCR detection of the methylenetetrahydrofolate reductase (MTHFR) C677T mutation. AB - BACKGROUND: Detection of the presence of 677C/T allele in the human MTHFR gene is easily achieved by amplification using primers designed to span this region. Hinf I digestion, occurring only in the 677T allele, subsequently discriminates between the two alleles. Existing methods, however, do not control for failed restriction endonuclease digestion. METHODS AND RESULTS: A new forward, modified primer was designed and placed further upstream to create a Hinf I site, which, because it is present in both alleles, would serve as an internal restriction control. CONCLUSIONS: By allowing an additional restriction to take place in both the normal and mutated alleles, the use of the new primer provided for an internal restriction control. PMID- 10462632 TI - Clinical news update PMID- 10462633 TI - Controversies in the prevention of neurosurgical infection. AB - Amongst post-operative infections, those associated with neurosurgery are of particular significance in view of their proximity to, or location within, the central nervous system. Superficial surgical site infections may be complicated by osteomyelitis of the calvarium and deeper extension to the meninges and cerebral parenchyma. The prevention, diagnosis and management of infections associated with implant devices provide similar challenges to those faced in orthopaedic and cardiac surgery. Whilst some consensus exists regarding the need for antimicrobial prophylaxis in the latter two disciplines, its place in neurosurgery remains controversial. When prophylaxis is considered, choice of antimicrobial agents should take account of up-to-date local information in relation to the relevant microbial ecology in hospitals and in the community setting. The potential for spread of blood-borne virus infections and transmissible spongiform encephalopathies in relation to neurosurgery must also be considered and it should be ensured that appropriate preventive strategies are in place. PMID- 10462634 TI - The role of the hospital environment in the epidemiology of multi-resistant bacteria. AB - The aim of this study was to use published data to assess the importance of the hospital environment as a possible secondary reservoir of multi-resistant bacteria capable of colonizing or infecting patients. This should make it possible to develop appropriate measures for preventing cross contamination in medical environments. Multi-resistant bacteria often contaminate the environment of the colonized or infected patients, and survive for long periods. However, measures proposed to reduce contamination and reduce the potential for cross infection such as disinfection when the patient is discharged, the use of materials with intrinsic antibacterial activity and the wearing of gloves by everyone entering the room whether or not they intend to actually touch the patient, have yet to be evaluated as part of an overall strategy to prevent infection with multi-resistant bacteria. PMID- 10462635 TI - The pros and cons of isolation and containment. AB - This paper is a review of isolation and containment models, policies and procedures. The arguments for and against various practices are outlined. The psychological effects of isolating patients in single rooms and new ideas for staff intervention are discussed. A modern, flexible approach is suggested. PMID- 10462637 TI - Rapid detection of methicillin-resistant staphylococci by multiplex PCR. AB - A multiplex PCR was developed to detect the coagulase gene (coa; pathognomic of Staphylococcus aureus) and the mecA gene (characteristically encoding for methicillin resistance in staphylococci) in a single, rapid test. Suitable primers for the gene targets and an internal, amplification control were incorporated into a multiplex PCR assay, which was then optimized on a capillary air thermal cycler to improve the turnaround time of the test to approximately 1.5 hours. The assay was evaluated with 111 fresh clinical isolates of staphylococci. The multiplex PCR correctly distinguished between isolates of S. aureus, which were sensitive to methicillin (MSSA) and those resistant to it (MRSA). It also correctly differentiated between similar isolates of coagulase negative staphylococci (MSSE and MRSE respectively). It was concluded that this multiplex PCR was a rapid and reliable method for the detection of methicillin resistant staphylococci. PMID- 10462636 TI - Detection of glycopeptide-resistant enterococci in routine diagnostic faeces specimens. AB - Faeces received in a diagnostic laboratory were screened for glycopeptide resistant enterococci (GRE) on modified Lewisham medium, with and without enrichment in Enterococcosel broth. Colonization by GRE was detected in 102/838 patients (12.2%). In 74 (73%) of colonized patients GRE were detected by both methods and in 28 (27%) they were detected only after enrichment. The carriage rate in hospitalized patients was 32% (93/289) compared with 2.3% (11/425) in the community (GP patients and food-handlers). Carriage of GRE increased with age. Clostridium difficile isolation was associated with GRE colonization, odds ratio 6.76 (P<0.001). Fifty-nine percent (60/102) of the GRE had the VanA phenotype and 41% (42/102) had the VanB phenotype. In the community VanA predominated (91%), whereas 64% (57/89) of the isolates from hospitalised patients were of the VanB phenotype. PMID- 10462638 TI - Molecular epidemiology of multi-resistant Escherichia coli. AB - In this case-control study multi-resistant Escherichia coli isolates were characterized on a molecular level and risk factors for their development were identified. Thirty-two multi-resistant E. coli strains were isolated from the urine of 13 patients attending a renal clinic for chronic urinary tract infection (UTI) and from different sites of 11 terminally ill patients with nosocomial infections hospitalized on five different wards. All 32 isolates were resistant to ciprofloxacin, cotrimoxazole and produced beta-lactamase. All strains contained plasmids of 2-110 MDa of which a 50 MDa and a 100 MDa plasmid were present in 81% of the strains. Pulse-field gel electrophoresis (PFGE) analysis demonstrated 17 genotypes among 32 strains which indicates a polyclonal outbreak with some geographic clustering. Monitoring of patients over the study period showed that either the resident genotype remained the same and that these retained strains underwent changes in their plasmid contents, or that they were replaced by a different genotype after several months of therapy for chronic UTI. Univariate analysis indicated that multi-resistant E. coli develop in the presence of long-term selective ciprofloxacin pressure at a dosing regimen of 250 mg bid for more than 20 days and that treatment with a broad spectrum antimicrobial for more than three days favours the selection of multi-resistant E. coli in the flora of terminally ill patients with multiple disorders. PMID- 10462639 TI - An analysis of hospital-acquired bacteraemia in intensive care unit patients in a university hospital in Kuwait. AB - An analysis of hospital-acquired bacteraemia among ICU patients was carried out over a two-year period in order to determine the incidence, associated mortality rate and susceptibility pattern of causative pathogens. There was a high incidence of bacteraemia, occurring in 127 (18.4%) of 692 patients. Mortality attributable to nosocomial bacteraemia was 52% of the total 79 deaths from all causes. The highest mortality rate (58.5%) occurred in patients with fungal infections, whilst death from Gram-negative bacteraemia was only 17%. Over 98% of patients had underlying disease. Nearly half (46.8%) of 267 organisms isolated were Gram-positive. In comparison, Gram-negative bacteria accounted for 36.6% and the rest (17.6%) were fungi (mainly Candida albicans). The majority of the bactereamic episodes were monomicrobial (90.2%). Coagulase-negative staphylococci (CNS) were the commonest pathogens isolated, representing 32.6% of all organisms. Inducible beta-lactamase producing organism (Enterobacter spp. 9.7%, Serratia marcescens 6.7%, Klebsiella pneumoniae 6% and Pseudomonas aeruginosa 6%) formed the bulk of Gram-negative bacteria. In contrast, Escherichia coli (7.5%) and K. pneumoniae (4%) were the commonest Gram-negative bacteria from hospital-acquired bacteraemia in the general hospital population. The majority (80%) of CNS were resistant to methicillin (MRSE) but susceptible to vancomycin; they were relatively resistant to erythromycin, clindamycin and beta-lactams antibiotics. Whilst Gram-negative organisms were relatively susceptible to imipenem (85%), ciprofloxacin (88%) and amikacin (87%), they had unacceptably low levels of susceptibility to cefuroxime (59.3%), cefotaxime (71%), ceftazidime (60.9%), and piperacillin (51.1%). This study shows that hospital-acquired bacteraemia in ICU patients carries a poor prognosis. Information regarding the infective agents and their susceptibility in the ICU setting is valuable for the selection of empirical therapy before culture and susceptibility results are known. PMID- 10462640 TI - Nosocomial transmission of hepatitis B virus infection through multiple-dose vials. AB - The source of acute hepatitis B virus (HBV) infection in two women (55 and 72 years old) was investigated. They displayed no risk factors for acquiring HBV infection, other than treatment with local anaesthetic injections some months previously. The HBV strains were sequenced and showed distinct homology to strains seen in Swedish intravenous drug users (IVDU). Prior to these patients' acute infection, an outbreak of HBV had occurred among IVDU in the same county. Analysis of the HBV strains from six of these IVDUs showed their core promoter, precore and pre-S sequences (679 nucleotides) to be identical to those from the two patients. Cross-contamination between samples was excluded and the most likely source of infection was thought to be multiple-dose vials of local anaesthetic that had been contaminated with the HBV strain circulating among the IVDU population in the community. We believe that multiple-dose vials have no place in modern healthcare and recommend sequence homology analysis as an alternative or additional way to trace a source of HBV infection. PMID- 10462641 TI - An outbreak due to echovirus type 30 in a neonatal unit in France in 1997: usefulness of PCR diagnosis. AB - Between February and August 1997, 53 patients with enterovirus meningitis were hospitalized in Clermont-Ferrand, France. All but one were children. Echovirus type 30 was involved in 70% of cases with identified serotype. The outbreak ceased on August 8. Two months later, a neonate was admitted to the neonatal unit with an echovirus type 30 meningitis thought to be acquired at delivery. Twenty days later a nosocomial outbreak of echovirus type 30 involving five neonates occurred. Two of them presented with meningitis and two with febrile seizure; One was asymptomatic. The retrospective examination of the maternal sera in a neutralization test, using the index case strain as a source of antigen, showed that none of the neonates was passively immunized before hospitalization. The use of genome detection in cerebrospinal fluid allowed rapid diagnosis and infection was contained by re-inforcing hygiene measures. Prospective examination of stools in the neonatal and paediatric units showed no further occurrences of the disease. No sporadic case was observed in the general population. Hence, nosocomial infections can occur a long time after an outbreak in the general population; rapid diagnosis with molecular tools is useful both for a definite diagnosis in patients already hospitalized, and to act as a rapid alert, even in intervals between seasonal outbreaks. PMID- 10462642 TI - A new method of assessing the penetration of bacteria through fabrics used in the operating theatre. PMID- 10462644 TI - Writing pens are an unlikely vector of cross-infection with methicillin resistant Staphylococcus aureus (MRSA) PMID- 10462643 TI - Identification of an IV-dextrose solution as the source of an outbreak of Klebsiella pneumoniae sepsis in a newborn nursery. PMID- 10462645 TI - Sepsis and ECMO: beware the breast milk. PMID- 10462648 TI - An initial experience with screening for colon polyps using spiral CT with and without CT colography (virtual colonoscopy) AB - BACKGROUND: Computed tomographic (CT) colography (virtual colonoscopy) is a new imaging method for detection of colon polyps and cancer. OBJECTIVE: To evaluate the sensitivity of CT colography for polyp detection in a population without symptoms that included persons without colon neoplasia and with radiologists blinded to colonoscopic findings. METHODS: Forty-six persons without symptoms underwent spiral CT followed by same-day colonoscopy with subsequent inspection of two-dimensional axial CT images, interactive multiplanar images, and surfaced and volume-rendered images of the colon (three-dimensional CT colography). RESULTS: Three-dimensional CT colography was superior to two-dimensional axial imaging for detection of colon polyps. Three-dimensional CT colography depicted 1 of 4 (25%) adenomas 2 cm in diameter or larger, 6 of 10 (60%) adenomas 1 to 1.9 cm, 6 of 14 (43%) 6 to 9 mm, and 7 of 65 (11%) 5 mm in diameter or smaller. Three dimensional CT colography showed a polyp that might have led to colonoscopy in 3 of 4 (75%) patients whose largest adenoma was 2 cm or larger, 5 of 6 (83%) patients with largest adenoma 1 to 1.9 cm, 3 of 7 (43%) patients with largest adenoma 6 to 9 mm, and 4 of 16 patients (25%) with largest adenoma 5 mm or smaller. Large, flat adenomas of the right colon were difficult to identify with three-dimensional CT colography. The specificity of three-dimensional CT colography for patients with adenomas 1 cm in diameter or larger was 89%. Examination of patients with missed adenomas after unblinding indicated that meticulous bowel preparation and adequate distention are critical to accurate interpretation. Perceptual errors were common. CONCLUSIONS: CT colography as performed in this study is not adequate as a colorectal cancer screening test. Several technical factors that appear critical to accurate performance of CT colography are defined. PMID- 10462649 TI - Do patients with rectosigmoid adenomas 5 mm or less in diameter need total colonoscopy? AB - BACKGROUND: The need for colonoscopy in the care of patients with rectosigmoid adenoma 5 mm or less in diameter is still debatable. METHODS: We estimated the prevalence of proximal adenomas among 3052 consecutive subjects undergoing total colonoscopy. Rectosigmoid adenoma was classified as diminutive (5 mm), small (6 to 10 mm), or large (>/=11 mm). Advanced proximal adenoma was 10 mm in diameter or larger, or with a villous component, severe dysplasia, or infiltrating adenocarcinoma. RESULTS: Proximal adenoma was found in 212 of 2483 patients (8.5%, 95% CI [7.5, 9.7]) without distal neoplastic polyps, 49 of 214 (22.9%, 95% CI [17.6, 29.2]) with diminutive, 44 of 174 (25.3%, 95% CI [19.1, 32.5] with small, and 70 of 181 (38.7%, 95% CI [31.6, 46.2]) with large distal adenoma. Advanced proximal adenoma was found in 49 (2.0%, 95% CI [1.5, 2.6]), 8 (3.7%, 95% CI [1.7, 7.5]), 17 (9.8%, 95% CI [6.0, 15.4]), and 29 patients (16.0%, 95% [11.2, 22.4]), respectively. In patients with distal adenoma risk for proximal lesions increased with increasing age, size, and number of distal adenomas (p = 0.01). Size of distal adenoma was the strongest predictor of the presence of proximal advanced adenoma (multivariate analyses). CONCLUSIONS: In a clinical setting, the decision to perform colonoscopy should take into account proximal lesions of clinical interest, life expectancy, costs, and risks associated with the procedure. When detection of advanced proximal adenoma is the goal, presence of distal diminutive adenoma alone might not be an indication for total colonoscopy. PMID- 10462650 TI - Prospective analysis of complications 30 days after outpatient colonoscopy. AB - BACKGROUND: Our objective was to (1) determine whether more complications are reported by patients 30 days after outpatient colonoscopy than are discussed at our monthly morbidity and mortality conferences, (2) identify complications resulting in visits to the emergency department or physician's office or leading to hospitalization, and (3) assess which factors put patients at highest risk. A secondary goal was to determine the rate of work lost after outpatient colonoscopy. METHODS: Trained interviewers performed standardized telephone interviews of consecutive outpatients undergoing colonoscopy at Georgetown University Hospital over a 1-year period. RESULTS: One thousand one hundred ninety-six patients were contacted 30 days after outpatient colonoscopy and participated in our study. Twenty patients had complications that required a visit to an emergency department or physician. Ninety percent of these cases (18) were detected at 30 days, but 15% (3) were discussed at morbidity and mortality conferences. All seven complications that necessitated hospitalization were identified at 30 days, but only two were discussed at our morbidity and mortality conference. The most common complications reported by patients were abdominal discomfort (5.4%) and rectal bleeding (2.1%). CONCLUSION: More complications are detected by means of contacting patients 30 days after outpatient colonoscopy than are discussed at our morbidity and mortality conferences. PMID- 10462651 TI - Correlation of lifting versus non-lifting and microscopic depth of invasion in early colorectal cancer. AB - BACKGROUND: The non-lifting sign is considered a contraindication to endoscopic resection. Our objective was to investigate whether lifting or nonlifting of a lesion is determined by the volume of normal submucosal tissue. METHODS: We measured the thickness of the submucosa and examined the relation between submucosal invasion and lesion elevation induced by submucosal injection in 60 patients with colorectal cancer with evidence of submucosal invasion. Extent of tumor elevation was classified into two groups: A, lifting; B, non-lifting. Submucosal invasion was classified as sm1, sm2, or sm3. The distance between the carcinoma and the line of resection and that between the carcinoma and the muscularis propria were measured. RESULTS: Of 31 sm1 lesions, 29 (93.5%) were group A. All 6 sm3 lesions were group B. All lesions in group A had a value for the distance between carcinoma and muscularis propria of more than 1000 microm. Group B lesions with sm3 invasion had distances of only 105 to 750 microm. CONCLUSION: Lesions classified as sm3 do not elevate in response to submucosal injection, and lesions that become elevated on injection can be resected endoscopically because they are sm1 or sm2 and have a thickness of normal submucosa of more than 1000 microm. PMID- 10462652 TI - Needle-knife fistulotomy versus needle-knife precut papillotomy for the treatment of common bile duct stones. AB - BACKGROUND: The aim of this prospective study was to evaluate and compare the efficacy and safety of two different precutting techniques in the treatment of 103 consecutive patients with choledocholithiasis. METHODS: The patients were randomized into two groups. The first group included 74 patients who underwent needle-knife fistulotomy avoiding the papillary orifice followed by standard papillotomy. Fifty-two of these patients had a final diagnosis of choledocholithiasis. The second group included 79 patients who underwent needle knife precut papillotomy starting from the papillary orifice followed by standard papillotomy. Fifty-one of these patients had a final diagnosis of choledocholithiasis. RESULTS: Precutting was successful in 90.54% of patients in the needle-knife fistulotomy group and 88.6% of patients in the needle-knife precut papillotomy group. Stone extraction without mechanical lithotripsy was achieved in 40 of 48 (83.33%) patients in the needle-knife fistulotomy group and 45 of 46 (97.82%) patients in the needle-knife precut papillotomy group (p < 0.05). For the other patients, stone extraction was achieved with the aid of a mechanical lithotriptor. Complications were as follows for the needle-knife fistulotomy and needle-knife precut papillotomy groups, respectively: bleeding, 6.75% and 5.06%; perforation, 2.7% and 2. 53%; cholangitis, 1.35% and 0; pancreatitis, 0 and 7.59% (p < 0.05); hyperamylasemia, 2.7% and 17.72% (p < 0.01); and death, 0 and 1.26%. CONCLUSIONS: Both methods are effective in the management of choledocholithiasis. When needle-knife fistulotomy is performed, however, lithotripsy is needed more often. Needle-knife fistulotomy is safer than needle-knife precut papillotomy with respect to pancreatic complications. PMID- 10462653 TI - ERCP and pyogenic liver abscess. AB - BACKGROUND: Liver abscess is commonly biliary in origin. We assessed the role of endoscopic retrograde cholangiopancreatography (ERCP) in patients with pyogenic liver abscesses. METHODS: Between January 1986 and December 1997, 63 patients with pyogenic liver abscesses were referred for ERCP. Twenty-one patients had a history of previous biliary procedures including cholecystectomy (21), biliary enteric bypass (9), surgical sphincteroplasty (5), and endoscopic sphincterotomy (2). Demographic data, clinical features, biochemical parameters, treatment, clinical progress, and follow-up were recorded and analyzed. RESULTS: Two patients required laparotomy and 61 patients underwent guided aspiration and/or drainage at a median interval of 3 days after presentation. ERCP was performed at a median of 8 days (range 1 to 69 days) after initial treatment and succeeded in 90% of cases with no associated complication or death. Abnormalities were shown in 29 (46%) patients: biliary obstruction due to stones or strictures (15), ductal dilatation alone (7), spontaneous choledochoduodenal fistula (3), communication between abscesses and biliary tree (3), and splaying of biliary ducts by space-occupying effect (2). No abnormality was found in 34 patients. Eight patients underwent endoscopic therapy including sphincterotomy (5), stone extraction (6), and nasobiliary drainage (2). Overall mortality rate from liver abscesses was 6%. CONCLUSIONS: ERCP is useful in the treatment of patients with pyogenic liver abscesses. PMID- 10462654 TI - Endoscopic therapy of fascioliasis resistant to oral therapy. AB - BACKGROUND: We present an endoscopic approach to patients with fascioliasis resistant to oral pharmacotherapy. A staged study was performed in which the effect of flushing the biliary system with a fasciolicidal solution was evaluated. METHODS: In phase I of the study, four agents (povidone iodine, potassium permanganate, chlorhexidine, and sodium bicarbonate) were tested in vitro for their effect on Fasciola worm viability. In phase II, patients resistant to oral pharmacotherapy for fascioliasis were referred for endoscopic retrograde cholangiopancreatography and flushing of the biliary system with the solution found to be most effective in phase I. RESULTS: Povidone iodine proved to be the most effective solution against Fasciola worm viability. Nine patients had the biliary system flushed with povidone iodine. The presence of a Fasciola worm was demonstrated in the bile duct of all patients. In one patient an extra worm was found in the gallbladder. All patients were negative for Fasciola ova on repeated follow-up stool examination. CONCLUSIONS: We recommend washing the biliary system of patients with fascioliasis resistant to oral pharmacotherapy with povidone iodine because it is effective against the worms in the bile ducts and gallbladder and usually leaves the patient with an intact papilla. PMID- 10462655 TI - Evaluation of metastatic celiac axis lymph nodes in patients with esophageal carcinoma: accuracy of EUS. AB - BACKGROUND: Endosonography (EUS) is the most accurate modality for assessing depth of tumor invasion and local lymph node metastasis. However, its accuracy in the identification of metastatic (celiac axis) lymph nodes is less well defined. Our objective in this study was to determine the accuracy of Eus in detecting celiac axis lymph node metastasis in patients with esophageal carcinoma. METHODS: Two hundred fourteen patients with esophageal carcinoma underwent preoperative EUS. Of these, 145 underwent attempted surgical resection and staging, and 4 underwent EUS-guided fine-needle aspiration of mediastinal and celiac lymph nodes. Local (mediastinal) and distant (celiac axis) lymph nodes were assessed for malignancy on the basis of four criteria (larger than 1 cm, round, homogeneous echo pattern, sharp borders). Accuracy of EUS was determined by means of correlating histopathologic findings for the resected lymph nodes or results of EUS-guided fine-needle aspiration cytologic examination. RESULTS: Surgical exploration (n = 145) and fine-needle aspiration cytologic examination (n = 4) revealed metastatic celiac axis lymph nodes in 23 and metastatic mediastinal (local) lymph nodes in 93 of 149 patients with esophageal carcinoma. According to defined criteria for malignant lymph nodes, there were 19 true-positive and 4 falsenegative results. Sensitivity for the diagnosis of celiac lymph node metastasis with EUS was 83% with a 98% specificity. For the diagnosis of mediastinal lymph node metastasis, sensitivity was 79% and specificity was 63%. All patients with malignant celiac axis lymph nodes had local T3 (tumor breaching adventitia) or T4 (tumor invading adjacent organs) disease. CONCLUSION: EUS is an excellent modality in the evaluation of metastatic celiac axis lymph nodes in patients with esophageal carcinoma. These findings should be used in selecting options for treatment. Sensitivity for detecting malignancy is consistent with that of prior studies, and local and regional lymph nodes and specificity is significantly higher. PMID- 10462656 TI - Endoscopic ultrasound (EUS) and EUS-guided fine-needle aspiration (FNA) of liver lesions. AB - BACKGROUND: Endoscopic ultrasonography (EUS) is not traditionally thought to be clinically applicable in liver imaging. EUS-guided fine-needle aspiration of the liver has not been well described. METHODS: A prospective study was conducted in which 574 consecutive patients with a history or suspicion of gastrointestinal or pulmonary malignant tumor undergoing upper EUS examinations underwent EUS evaluation of the liver. Fourteen (2.4%) patients were found to have focal liver lesions and underwent EUS-guided fine-needle aspiration. RESULTS: The median largest diameter of the liver lesions was 1.1 cm (range 0.8 to 5.2 cm). The mean number of passes was 2.0 (range 1 to 5 passes). All fine-needle passes yielded an adequate specimen. One of the 14 patients underwent EUS-guided fine-needle aspiration of 2 liver lesions. Fourteen of the 15 liver lesions sampled by means of EUS-guided fine-needle aspiration were malignant and one was benign. Before EUS, computed tomography (CT) depicted liver lesions in only 3 of 14 (21%) patients. Seven of 14 patients had a known cancer diagnosis. For the other 7, the initial diagnosis of cancer was made by means of EUS-guided fine-needle aspiration of the liver. There were no immediate or late complications. CONCLUSIONS: EUS can detect small focal liver lesions that are not detected at CT. Findings of EUS-guided fine-needle aspiration can confirm a cytologic diagnosis of liver metastasis and establish a definitive M stage that may change clinical management. PMID- 10462657 TI - GI endoscopic reprocessing practices in the United States. AB - BACKGROUND: Patient infection from contaminated gastrointestinal (GI) endoscopes can generally be attributed to failure to follow appropriate reprocessing guidelines. Recently, the Food and Drug Administration recommended a 45-minute exposure of GI endoscopes to 2.4% glutaraldehyde solutions heated to 25 degrees C. Simultaneously, the American Society for Gastrointestinal Endoscopy (ASGE), the American Gastroenterological Association, and the Society of Gastroenterology Nurses and Associates endorsed a reprocessing guideline that emphasized manual precleaning and recommended a 20-minute exposure to a 2.4% glutaraldehyde solution at room temperature. Since then, little information has become available regarding actual reprocessing practices in the United States. METHODS: A previously developed questionnaire regarding endoscopic disinfection practices was mailed to randomly selected members of the ASGE. RESULTS: The survey was sent to 730 members and 294 responded (40.3%). Appropriate manual cleaning (suctioning detergent through the accessory channel and brushing the channel and valves) is performed by 90.7% of respondents; 69.9% then use automated reprocessors for disinfection or sterilization. Glutaraldehyde is the most widely used chemical disinfectant; 85.3% use glutaraldehyde as one of their primary disinfectants. The most commonly used disinfection time with 2.4% glutaraldehyde is 20 minutes (83.9%) followed by 45 minutes (11.4%). Only 23.8% of users of 2.4% glutaraldehyde heat their solution; 59.6% of centers test disinfectant concentration daily or more frequently; 74.0% sterilize nondisposable forceps before use; 29.2% of centers re-use disposable endoscopic accessories (which are more frequently disinfected rather than sterilized). Twelve respondents reported cases of endoscopic cross infection. CONCLUSIONS: A significant minority of endoscopy centers still do not completely conform to recent ASGE, American Gastroenterological Association, and the Society of Gastroenterology Nurses and Associates guidelines on disinfection, and they may not be appropriately disinfecting GI endoscopes. Rigid adherence to recommended guidelines is strongly encouraged to ensure patient safety. PMID- 10462658 TI - Comparison of endoscopic variceal sclerotherapy with sequential endoscopic band ligation plus low-dose sclerotherapy for secondary prophylaxis of variceal hemorrhage: a prospective randomized study. AB - BACKGROUND: Endoscopic variceal sclerotherapy and band ligation both have certain limitations such as, respectively, esophageal complications and early recurrence of varices. METHODS: From February 1994 to March 1996, all consecutive patients with portal hypertension due to either cirrhosis or noncirrhotic portal fibrosis and a history of variceal bleeding were included in a prospective study and randomly assigned to receive either endoscopic variceal sclerotherapy alone or endoscopic variceal band ligation plus low-dose endoscopic variceal sclerotherapy. RESULTS: Of 69 patients, 34 were randomly assigned to receive endoscopic variceal sclerotherapy alone; 35 received endoscopic variceal band ligation plus endoscopic variceal sclerotherapy. Complete variceal eradication rates (85% vs. 80%) and the number of endoscopic sessions required for eradication (6.61 +/- 2.94 vs. 7.85 +/- 3.31) were similar in the endoscopic variceal sclerotherapy and endoscopic variceal band ligation plus endoscopic variceal sclerotherapy groups, respectively. The mean volume of sclerosant required in the combined group (54.94 +/- 33.74 mL) was significantly less than that in the endoscopic variceal sclerotherapy group (81.91 +/- 34.80 mL). The complication and recurrent bleeding rates were significantly higher in the endoscopic variceal sclerotherapy group than those in the combined group (20% and 16% vs. 3% and 3%, respectively). CONCLUSIONS: Both endoscopic variceal sclerotherapy and endoscopic variceal band ligation plus endoscopic variceal sclerotherapy were comparable in eradicating varices but the combined technique was associated with significantly lower complication and recurrent bleeding rates. PMID- 10462659 TI - EUS analysis of collateral veins inside and outside the esophageal wall in portal hypertension. AB - BACKGROUND: The relation between esophageal varices and venous collaterals outside the esophageal wall in patients with portal hypertension remains unclear. We investigated this relationship with endoscopy and endoscopic ultrasound (US). METHODS: Twenty-two patients with untreated varices were examined. The collaterals were studied with a 20 MHz US catheter probe. Collaterals were divided into two groups: (1) periesophageal collateral veins adjacent to the muscularis externa of the esophagus and (2) paraesophageal collateral veins external to the esophageal wall but without contact with the muscularis externa. Periesophageal and paraesophageal collateral veins were scored as mild or severe according to stage of development. RESULTS: Varix form as defined endoscopically was significantly larger in the severe periesophageal collateral veins group than in the mild periesophageal collateral veins group (p < 0. 01). In contrast, varix form did not differ significantly between the mild and severe paraesophageal collateral veins groups. Perforating veins were detected in the distal esophagus in 18 of 22 patients (81.8%) by means of US. The prevalence of perforating veins increased in relation to varix form. Two types of perforating veins were found on the basis of connection with periesophageal and paraesophageal collateral veins. The frequency with which perforating veins were connected to periesophageal collateral veins was 81.8%; the frequency of connection to paraesophageal collateral veins was 27.3%. CONCLUSION: Periesophageal collateral veins play a more important role in the formation of esophageal varices than do paraesophageal collateral veins. PMID- 10462660 TI - Does quantitative serologic testing for Helicobacter pylori predict peptic ulcer disease in cirrhosis? AB - BACKGROUND: Helicobacter Pylori is strongly associated with peptic ulcer disease. A correlation between high IgG serum antibody concentrations and a clinical diagnosis has been reported. It has been recognized that peptic ulcer disease occurs with increased frequency in cirrhosis. In this study, we attempted to establish a relation between the magnitude of serum IgG antibody to H pylori and the endoscopic diagnoses in H pylori-infected cirrhotic patients. METHODS: All cirrhotic patients who had undergone esophagogastroduodenoscopy with a positive H pylori IgG serology and who did not receive anti-H pylori treatment or take medications noxious to the gastroduodenal mucosa were included in the study. H pylori IgG serology was determined by an enzyme-linked immunosorbent assay with values of greater than 50 units/mL being seropositive. The functional reserve of cirrhosis was classified by modified Pugh-Child criteria. RESULTS: One hundred four seropositive cirrhotic patients were recruited. The serum IgG titers ranged from 51 to 1200 units/mL with a peak frequency at 50 to 99 units/mL (35. 6%). Statistical analysis did not reveal any relation between the quantitative H pylori IgG values and the endoscopic diagnoses, which included gastric ulcers, duodenal ulcers, gastroduodenal erosions, and normal findings. CONCLUSIONS: In cirrhosis, the magnitude of H pylori IgG serology cannot be used to predict the presence or absence of peptic ulcer disease. PMID- 10462661 TI - Gastrocolic fistula in Crohn's disease. PMID- 10462662 TI - A high-intensity US probe designed for intraductal tumor destruction: experimental results. AB - BACKGROUND: Many digestive tract tumors spread inside the lumen and are not amenable to curative surgical treatment. An intraluminal method of tumor destruction would be useful for palliative or even curative purposes. High intensity ultrasound (US) is suitable for such purposes. Our objective was to perform experiments with animal models that would lead to development of a high intensity US probe for intraductal tumor destruction suitable for insertion through a large-channel endoscope. METHODS: The active part of the high-intensity US applicator consisted of a water-cooled piezoceramic plane transducer (3 x 10 mm) operating at 5 MHz for deep or 10 MHz for shallow tissue penetration. A cylinder of tissue was destroyed by means of rotating the transducer on its axis through a flexible shaft. Experiments were conducted in vitro on livers of butchered pigs (10 lesions), in vivo on exteriorized pig livers (15 lesions), and on metastatic Dunning tumors (AT(2 ) subline) implanted subcutaneously in 28 rats (treated n = 16, controls n = 12). RESULTS: In experiments on pig livers, high intensity US induced highly reproducible cylinders of coagulation necrosis (diameter 20 +/- 1 mm, height 8 +/- 1 mm) with sharply demarcated and serrated boundaries. The exposure duration to achieve such lesions was 5 minutes. Regions of coagulation necrosis obtained in vivo were similar in size and shape. All 12 control rats died or were killed because of diffuse cancer by day 15 after implantation; 64% of the treated rats were tumor free 30 days after treatment, and 36% had local recurrences. CONCLUSION: This high-intensity US probe induces highly reproducible cylinders of coagulation necrosis and is effective against tumors in animals. PMID- 10462663 TI - EUS-guided radiofrequency ablation in the pancreas: results in a porcine model. AB - BACKGROUND: Our aim in this study was to investigate the feasibility and safety of performing radiofrequency (RF) ablation in the pancreas with endoscopic ultrasound (EUS). METHODS: RF was applied to normal pancreatic tissue in 13 anesthetized Yorkshire pigs with specially modified 19-gauge needle electrodes (1.0 to 1.5 cm tip). The pancreas was localized with EUS and punctured through a transgastric approach. RF current (285 +/- 120 mA) was delivered for 6 minutes. Diagnostic imaging (EUS and CT) and serum amylase and lipase levels were obtained at baseline, immediately after ablation, and 1 to 14 days after the procedure. Pigs were killed immediately (n = 5), 1 to 2 days after ablation (n = 2), and 2 weeks after the procedure (n = 6). Pathologic examination was performed. RESULTS: Sixteen ablations were performed. During ablation, round hyperechoic foci (diameter to 1.0 cm) gradually surrounded the tip of the electrode. Immediately after the procedure CT demonstrated 1 cm hypodense foci that did not enhance with iodinated contrast. In pigs killed immediately and 1 to 2 days after ablation, pathologic examination showed discrete, well-demarcated spherical foci of coagulation necrosis measuring 8 to 12 mm in diameter surrounded by a 1 to 2 mm rim of hemorrhage. Radiologic-pathologic correlation was within 2 mm. In 4 of 6 (67%) pigs killed on day 14, retraction of the coagulated focus was observed. A 1 to 3 mm fibrotic capsule surrounded the coagulated tissue in the remaining 2 pigs. One pig had mild hyperlipasemia, a focal zone of pancreatitis (<1 cm), and later a pancreatic fluid collection. Biochemical parameters were normal in the remaining pigs. Other complications included three gastric and one intestinal burn caused by improper electrode placement. CONCLUSIONS: EUS-guided RF ablation can be used safely to produce discrete zones of coagulation necrosis in the porcine pancreas. Potential clinical uses of this technology include management of small neuroendocrine tumors and possibly palliation of unresectable pancreatic adenocarcinoma. PMID- 10462664 TI - New wire-guided basket for intrahepatic stone extraction. AB - BACKGROUND: Endoscopic extraction of intrahepatic ductal stones with conventional stone retrieval baskets sometimes is difficult when the stones are deep in the segmental ducts or in a tortuous duct. We evaluated the use of a new wire-guided basket for endoscopic bile duct stone extraction. METHODS: The new wire-guided basket has a separate lumen for the guidewire on one side of the catheter. It is rail-loaded and advanced over a guidewire into the desired segmental duct. After they are engaged, the stones are dragged into the duodenum and the guidewire is left in the segmental duct. The basket then can be reinserted into that particular ductal segment over the wire for further stone extraction. RESULTS: The basket was used to treat three patients. Two patients had multiple intrahepatic stones. Repeated passage of the basket back to the desired location was accomplished without difficulty. All stones were removed successfully. CONCLUSION: The new wire-guided basket is a useful device for removing intrahepatic stones, particularly from patients with multiple ductal stones. PMID- 10462665 TI - Endoclip-assisted resection of large pedunculated colon polyps. AB - BACKGROUND: Large pedunculated colorectal polyps are often difficult to remove endoscopically. METHODS: Four patients with giant pedunculated polyps (>3 cm) underwent an endoscopic procedure in which the polyp stalk was ligated with multiple clips and resected with a needle-knife papillotome. RESULTS: No complications occurred either during or immediately after the procedure. Mean size of the resected polyps was 4.8 +/- 1.2 cm (range 3 to 6 cm). Mean time for the entire procedure was 31 +/- 5 minutes (range 25 to 45). No bleeding was found after a mean follow-up period of 4.3 +/- 1.1 months (range 3 to 6 months). CONCLUSIONS: Although technical refinements are necessary to make the procedure easier and faster, this approach might represent the only option for selected patients who would otherwise need surgical resection of large pedunculated colonic polyps. PMID- 10462666 TI - Esophagopleural fistula treated endoscopically with argon beam electrocoagulation and clips. PMID- 10462667 TI - Nonsurgical treatment of duodenal perforation by endoscopic repair using a clipping device. PMID- 10462669 TI - Combined laparoendoscopic cystgastrostomy. PMID- 10462668 TI - Simultaneous mucosa-associated lymphoid tissue lymphoma of the stomach and colon. PMID- 10462670 TI - Hemobilia in advanced pancreatic cancer with portal vein obstruction and a metal endobiliary stent: a case report. PMID- 10462671 TI - Treatment of a radiation-induced sigmoid stricture with an expandable metal stent. PMID- 10462672 TI - Self-expanding metallic stents for palliation of recurrent malignant esophageal obstruction after subtotal esophagectomy for cancer. PMID- 10462673 TI - Endoscopic clip-assisted biliary cannulation: externalization and fixation of the major papilla from within a duodenal diverticulum using the endoscopic clip fixing device. PMID- 10462674 TI - Endoscopic removal of rectal leiomyoma: case report. PMID- 10462675 TI - Appendicular ascariasis: colonoscopic management. PMID- 10462676 TI - Screening by CT colonography: too early to pass judgment on a nascent technology. PMID- 10462677 TI - Endoclip: closing the surgical gap. PMID- 10462678 TI - The OMED Colorectal Cancer Screening Committee: a report of its aims and activities. Organisation Mondiale d'Endoscopie Digestive. PMID- 10462679 TI - Endoscopic band ligation for actively bleeding Dieulafoy's lesions. PMID- 10462680 TI - Pancreatic pseudocyst in a patient with cirrhotic ascites and portal hypertension: effective endoscopic drainage with Seldinger technique. PMID- 10462681 TI - Recurrent GI hemorrhage following the use of silk sutures. PMID- 10462682 TI - Painless removal of traction-removable PEG tubes. PMID- 10462683 TI - Establishment and plasticity of neuronal polarity. AB - A fundamental feature of neurons is that they possess a polarized morphology, typified by a single long axon and several short dendrites. This cellular polarity forms the basis for directionalized rapid signaling, and for bi directional trophic signaling, in neuronal circuits. While a catalog of structural, molecular, and functional differences between axons and dendrites is accumulating, the mechanisms involved in the establishment of neuronal polarity (also referred to as axogenesis) are not well understood. Cytoskeletal components, including microtubules and actin filaments, and cytoskeleton regulating proteins such as microtubule-associated proteins (MAPs), actin-binding proteins, and "motor" proteins, likely play important roles in the establishment and maintenance of neuronal polarity. Polarized sorting of membranes and proteins occurs in neurons, and a better understanding of such sorting mechanisms will likely provide insight into the process of axogenesis. Calcium plays a pivotal role in the regulation of neurite elongation and growth cone motility. Studies in which intracellular calcium levels are measured and manipulated in embryonic neurons during the process of axogenesis suggest a necessary role for calcium gradients in the establishment of neuronal polarity. Experimental models in which developing neurons are induced to switch their polarity support central roles for calcium-regulated cytoskeletal reorganization and protein sorting in the process of axogenesis. Finally, recent findings suggest that mitochondria play an important role in organizing neuronal polarity, possibly by controlling local calcium and/or energy gradients. While considerable progress is being made in elucidating mechanisms that regulate neuronal polarity, the seminal event(s) underlying this process remain a mystery. J. Neurosci. Res. 57:577-589. PMID- 10462684 TI - Lineage restriction of neuroepithelial precursor cells from fetal human spinal cord. AB - In the presence of epidermal growth factor (EGF) and/or fibroblast growth factor 2 (FGF2), neuroepithelial precursor cells from dissociated fetal human spinal cord are mitotically active and form free-floating spheres of undifferentiated cells. Proliferating cells were obtained in approximately 40% of preparations with each mitogen, were immunoreactive for the intermediate filament nestin, and did not express neuronal- or glial-specific markers. Early passage neuroepithelial precursor cells were pluripotent and differentiated into neurons expressing MAP2a,b, NF-M, and TuJ1, and GFAP-positive astrocytes; however, oligodendrocytes were never seen. As the cells were passaged from P0 to P4, the percentage of differentiating neurons significantly decreased and the prevalence of astrocytes significantly increased. While the majority of cell populations from individual preparations stopped proliferating between 3 and 6 passages, two expanding cell lines have been successfully expanded in EGF and FGF2 for over 25 passages and have been maintained in culture for over one year. These cells express nestin and not other cell-specific lineage markers. When differentiated, these neuroepithelial cell lines differentiate only into astrocytes, showing no expression of any neuronal marker. These data suggest that continued passage under these conditions preferentially selects for spinal cord neural precursors that are restricted to the astrocytic lineage. Despite the lineage restriction of later passage cell populations, these results provide a rationale for future investigation into the lineage potential of these cells in vivo following transplantation into the adult CNS, potentially as a therapeutic approach for traumatic injury and neurodegenerative disease. PMID- 10462685 TI - Differential expression of estrogen receptor alpha and beta in rat dorsal root ganglion neurons. AB - Estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta) mRNAs are both expressed in rat dorsal root ganglion (DRG) neurons, but the distribution of these two mRNAs differs markedly. Radiolabeled probes highly specific to ERalpha or ERbeta mRNAs were used for in situ hybridization studies; two antibodies specific to ERalpha protein were used for immunocytochemistry and specific primers were used for reverse transcription polymerase chain reaction (RT-PCR) studies. These revealed that ERbeta mRNA is widely expressed in the DRG of both male and female rats, with virtually all neurons showing positive signals. In contrast, ERalpha mRNA, as well as nuclear localized ERalpha protein, is more restricted in its localization and is present in many, but not all, of the small sized (<600 microm(2)) DRG neurons, but is only rarely present in larger neurons. The L6-S1 DRG levels, which contain sensory neurons that innervate reproductive tissues, are relatively enriched in ERalpha compared to L3-L5 DRG levels, which contain sensory neurons that innervate hind limb regions. Long-term estrogen treatment of ovariectomized rats (21-28 days) dramatically reduces immunocytochemically detectable ERalpha protein in the DRG relative to that in ovariectomized controls. RT-PCR studies also showed that long-term estrogen treatment of ovariectomized rats downregulates the levels of ERalpha mRNA, but upregulates the levels of ERbeta mRNA in the DRG. Interestingly, in intact cycling female rats, ERalpha and ERbeta mRNA levels in the DRG were both higher during proestrus compared to metestrus. These findings suggest that the changes in expression of estrogen receptors which occur dynamically during the estrus cycle differ from those induced by long-term estrogen treatment of ovariectomized animals. PMID- 10462686 TI - Expression of colony stimulating factor-1 receptor (CSF-1R) by CNS neurons in mice. AB - We report that neurons in the central nervous system express colony stimulating factor-1 receptor (CSF-1R) mRNA and protein and that the expression has regional specificity. The presence of CSF-1R in neurons was demonstrated by the use of four different types of antibodies to CSF-1R and the presence of CSF-1R mRNA by in situ hybridization using oligonucleotide probe. In the steady state in most areas of the brain, CSF-1R is weakly expressed in only a few neurons. In the cerebellum, brainstem, and spinal cord, however, CSF-1R is expressed constitutively in greater numbers of neurons. After cerebral cortex ischemic injury, neurons in the area next to the ischemic lesion markedly upregulate CSF 1R. It is also upregulated in the contralateral cortex and in many other areas of the brain and spinal cord. We demonstrated that in cultures the ligand CSF-1 binds to its receptor (CSF-1R) in neurons and that reduction of the number of apoptotic neurons and potentiation of neuron survival is CSF-1 dose dependent. We propose that CSF-1/CSF-1R signaling is an important regulatory pathway between neurons, microglia, and astrocytes. PMID- 10462687 TI - Increased intracellular calcium alters myelin gene expression in the N20.1 oligodendroglial cell line. AB - Regulation of intracellular Ca(2+) (Ca(i)) plays a central role in cell survival, proliferation, and differentiation. We previously reported that immature oligodendroglia (OLs) are less susceptible than mature OLs to cell death following increases in Ca(i) (Benjamins and Nedelkoska [1995] Neurochem. Res. 21:471-479). The N20.1 murine OL cell line provides a model of an intermediate stage of OL maturation in which to study responses to Ca(i) increases with regard to viability, as well as the expression of mRNAs for myelin basic protein (MBP), proteolipid protein (PLP), DM-20, SCIP, and the immediate early genes ZIF268, c fos, and c-jun. Cells were treated with the calcium ionophore A23187 or thapsigargin for 1, 3, and 18 hr. A23187 at 1.0 microM had no significant effect on cell detachment or death, whereas thapsigargin at 1.0 microM slightly increased both. With both agents, SCIP, MBP, and PLP mRNA levels were unaffected by 3 hr, but markedly reduced after 18 hours. DM-20 mRNA levels remained unchanged at both time points. With both agents, ZIF268, c-fos, and c-jun mRNA levels were unaffected after 1 hr; c-jun mRNA levels showed a significant increase after 3 hr of thapsigargin treatment. Thus, in N20.1 cells, increased calcium affects the IEG c-jun first, SCIP is coordinately decreased with MBP and PLP mRNAs at a later time point, and DM-20 message is under different regulation than PLP. J. Neurosci. Res. 57:633-642. PMID- 10462688 TI - Stimulus-secretion coupling in porcine adrenal chromaffin cells: acute effects of glucocorticoids. AB - Recent studies from this laboratory have established that long-term exposure (48 hr) to glucocorticoids can modulate voltage-gated Ca(2+) channel activity and subsequent intracellular Ca(2+) transients in porcine adrenal medullary chromaffin (PAMC) cells maintained in primary culture. Consistent with many steroid hormone-mediated responses, this chronic effect of glucocorticoids probably involves increased gene expression and protein synthesis. However, there is now considerable evidence to suggest that steroids can also elicit acute, non genomic effects. The aim of the present study was to determine whether acute exposure to glucocorticoids also affects nicotinic receptor-dependent catecholamine (CAT) secretion and Ca(2+) signaling in PAMC cells. Acute exposure to dexamethasone (DEX) dose-dependently attenuated the degree of nicotine (NIC) induced CAT secretion, as well as the amplitude of NIC-induced intracellular Ca(2+) transients. Significant inhibition of CAT secretion occurred immediately upon addition of DEX, reached maximal levels within 5 min of exposure to DEX, and was rapidly reversible after steroid washout. The endogenous porcine glucocorticoid cortisol elicited similar effects. In contrast, DEX had no significant effect on KCl-induced CAT secretion or intracellular Ca(2+) transients. These data demonstrate that acute exposure to glucocorticoids can modulate stimulus-secretion coupling in PAMC cells and suggest that the primary site of action is the nicotinic receptor. PMID- 10462690 TI - Inhibition of high glucose-induced protein mono-ADP-ribosylation restores neuritogenesis and sodium-pump activity in SY5Y neuroblastoma cells. AB - The exposure of SY5Y neuroblastoma cells to high concentrations of glucose, fructose, or galactose is an experimental model commonly used for in vitro evaluation of typical neuronal alterations observed in diabetes mellitus. In the present study, we observed that 2 weeks of exposure to high carbohydrate concentrations caused both a significant impairment in neurite formation induced by supplementation of retinoic acid or by subtraction of fetal calf serum to the culture medium and a marked reduction in Na(+)-K(+)-ATPase activity. However, only the exposure to high millimoles of glucose caused an enhancement of mono-ADP ribosylation, typical of diabetes mellitus, affecting at least five proteins. The concomitant exposure to high glucose and to silybin, a mono-ADP-ribosylation inhibitor, normalized the extent of ADP-ribosylation of the five proteins and counteracted the inhibitory effects of high glucose on Na(+)-pump activity and on neuritogenesis. Conversely, the supplementation of silybin did not prevent fructose and galactose inhibitory effects on Na(+)-pump activity and neurite formation. These data confirm those of previous reports suggesting a link between excessive protein mono-ADP-ribosylation and the onset of diabetic complications such as diabetic neuropathy. PMID- 10462689 TI - Comparative study of the distribution of calmodulin kinase II and calcineurin in the mouse brain. AB - The Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and the phosphatase calcineurin (CaN) are Ca(2+)/calmodulin-binding proteins that are very abundant in the central nervous system. In the mammalian brain, CaMKII is composed by the association of several similar subunits at different ratios produced by four different genes. CaN is composed of two different subunits produced by two genes. We selected the most abundant subunits of each enzyme in the rodent brain, CaMKII alpha and CaN A, and compared their pattern of expression in the mouse brain by using in situ hybridization histochemistry and immunohistochemistry. We found that CaMKII and CaN were mainly expressed in cerebral cortex, hippocampus, and striatum and that low levels of expression were observed in midbrain and brainstem. CaN was also expressed in cerebellum. In the cell, the kinase and the phosphatase were detected in the perikarya, the neuronal processes, and the nucleus. The present study shows that all the regions of the mouse brain in which CaMKII is expressed also show CaN expression. This fact is consistent with the presence of common substrates for both enzymes or with a regulatory action of one versus the other. The lack of correspondence in the cerebellum could be explained by the fact that the major subunit of the kinase in this brain region is CaMKII beta. PMID- 10462691 TI - Isolation and characterization of unmyelinated axolemma from bovine splenic nerve. AB - Bovine splenic nerve was used as a source of axolemma-enriched fractions derived from mammalian unmyelinated axons. By electron microscopy, splenic nerve consisted entirely of fascicles of unmyelinated axons and associated Schwann cells. The epineurium and blood vessels were stripped from the dissected nerve, which was then homogenized followed by preparation of a microsomal fraction by differential centrifugation. The microsomes were fractionated on a 10% to 40% continuous sucrose gradient. The individual fractions were combined into six fractions based on sucrose concentration and each fraction was analyzed for membrane markers. The 20% to 23% region of the sucrose gradient was enriched approximately sevenfold in acetylcholinesterase activity and twofold enrichment in saxitoxin binding activity was noted in the same fraction. Relative to other microsomal fractions, this same fraction was less enriched in a microsomal marker (cytochrome c reductase) and only moderately enriched in the activity of a myelin membrane marker (2',3' cyclic nucleotide 3' phosphohydrolase, CNPase). Polyacrylamide electrophoresis of the axolemma-enriched fraction revealed five prominent peptides ranging in molecular weight from 40 kDa to 130 kDa. Lipids, comprising 59.4% of the dry weight, were enriched in cholesterol and sphingomyelin, consistent with the origin from a peripheral nervous system (PNS) plasma membrane. On a molar basis, the major gangliosides were G(T1b), G(D1a), and G(M1). As a whole, these molecular characteristics are consistent with the origin of the axolemma-enriched fraction in the unmyelinated splenic nerve axons. This membrane preparation should prove useful in future studies of the myelinogenic potential of mammalian unmyelinated axolemma. PMID- 10462692 TI - Sympathectomy decreases and adrenergic stimulation increases the release of tissue plasminogen activator (t-PA) from blood vessels: functional evidence for a neurologic regulation of plasmin production within vessel walls and other tissue matrices. AB - Our recent morphologic studies indicated that peripheral nervous system (PNS) adrenergic neurons synthesize, transport, and store the serene protease, tissue plasminogen activator (t-PA) in axon terminals, many of which innervate vessel walls. Sympathoadrenal stimulation induces a surge of t-PA from vessel walls into the blood. The vascular endothelium, which constitutively secretes t-PA into blood also has long been widely assumed to be the principal source of this stress induced release, but has not been verified as such. A neurologically regulated release from adrenergic stores could thus augment the known constitutive endothelial release. To functionally test this possibility, we quantitated the effects of guanethidine-induced systemic sympathectomy on the basal and stimulated release of t-PA from isolated vessel explants in superfused organ cultures. Moment-to-moment changes in the release rate were plotted from serial assays of the t-PA free activity. The effects of endothelial and adventitial nerve plexus ablations were also tested. Sympathectomy induced 30-50% reductions in t-PA release from both arterial and microvascular explants. An acute release induced by alpha-1 adrenergic receptor stimulations was also strongly suppressed, as were basal levels of the circulating enzyme in vivo. Adventitial and endothelial ablations from normal large vessel explants produced greater reductions than small vessel endothelial ablations. Ganglion electrical stimulation also induced an acute microvascular release in vivo. These and past morphologic findings indicate a physiological infusion of t-PA into the vessel walls, blood, and other innervated matrices by sympathetic neurons. PMID- 10462693 TI - Insulin-sensitive GLUT4 glucose transporters are colocalized with GLUT3 expressing cells and demonstrate a chemically distinct neuron-specific localization in rat brain. AB - The insulin-sensitive glucose transporter (GLUT) 4, expressed primarily in peripheral tissue, has recently been detected also in the brain, demonstrating a region-specific distribution. To identify the chemical nature of neurons expressing GLUT4 and to disclose whether GLUT4-containing neurons also express the GLUT3 isoform, combined in situ hybridization for GLUT3 mRNA and double labeling immunocytochemistry for GLUT4 and different cellular markers was performed in brain sections through rat basal forebrain, cerebral cortex, hippocampus, and cerebellum. In all brain regions examined, GLUT4 immunoreactivity was exclusively found in neurons, and GLUT4-immunoreactive cells were colocalized with neurons expressing GLUT3 mRNA. In rat basal forebrain, cholinergic and parvalbumin-containing gamma-aminobutyric acid-ergic cells demonstrated GLUT4 immunoreactivity, whereas calretinin-, calbindin-D-, and neuronal nitric oxide synthase-containing neurons did not express GLUT4 protein. Because brain GLUT4 transporters have been suggested to play a role in rapidly providing additional glucose to neurons under conditions of high-energy demand, the selective presence of GLUT4 in basal forebrain cholinergic cells may explain the specific vulnerability of these cells to a lack of glucose supply. PMID- 10462695 TI - MK-801-induced expression of Fos protein family members in the rat retrosplenial granular cortex. AB - The N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 has been shown to induce an acute Fos and Fos-related antigen (Fra) expression in the rat retrosplenial granular cortex (RSG), but the exact composition of the Fos protein family and their individual dynamic alterations are unknown. We examined this issue using immunocytochemistry and Western blot analysis with two antibodies that recognize, respectively, Fos and all the identified members of Fos protein family. Immunocytochemistry detected a rapid and transient expression of Fos proteins in some RSG neurons and a delayed and prolonged expression of Fra proteins in most RSG neurons following a single systemic MK-801 injection (1 mg/kg). Multiple MK-801 injections (i.e., ten consecutive injections once every other day; 1 mg/kg) produced a moderate Fra expression but failed to induce detectable Fos expression. Western blot analysis further showed a transient expression of 72-kDa Fos proteins following a single MK-801 injection and a prolonged expression of 46- and 43-kDa Fra proteins after either a single or multiple MK-801 administration. The delayed onset and prolonged expression of these Fra proteins suggest that they may be DeltaFosB. The possible relevance of these results to clinical psychotomimetic effects of the NMDA receptor antagonists phencyclidine and ketamine is discussed. PMID- 10462694 TI - Anti-recoverin antibodies cause the apoptotic death of mammalian photoreceptor cells in vitro. AB - We developed an in vitro model to study the effect of anti-recoverin antibodies on retinal cells and the mechanism(s) by which they kill photoreceptors in cancer associated retinopathy (CAR). Rat retinal cells were grown in a defined medium, and cell types were identified by using antibodies against rhodopsin, recoverin, syntaxin, and thy-1. Purified immunoglobulin (IgG) against recoverin was added to the cultures at different concentrations for 24, 48, or 72 hr, and the survival of the cells was determined by fluorescence microscopy. Preimmune IgG and normal medium were used as controls. The cell death detection enzyme-linked immunosorbent assay and the terminal deoxyuridine triphosphate nick-end labeling assay were used to demonstrate cells undergoing apoptosis. Double labeling was used to visualize cell types and apoptotic death. Rods, amacrine cells, and ganglion cells were identified in the cultures. Rod cells, but not ganglion cells and amacrine cells, markedly decreased in the presence of 200 microg/ml of anti recoverin IgG for 24, 48, and 72 hr. Anti-recoverin antibodies caused apoptosis in rod cells but not in amacrine cells. Almost all cells were shown to take up IgG from the medium. In conclusion, our retinal cell cultures provide a system for investigating antibody-mediated photoreceptor cell death and demonstrate that anti-recoverin antibodies cause the apoptotic death of rod cells, with no effect on amacrine cells. The results suggest that anti-recoverin antibodies play a key role in the apoptotic death of photoreceptors in CAR. PMID- 10462696 TI - Expression of the beta-trace protein in human pachymeninx as revealed by in situ hybridization and immunocytochemistry. AB - In the pachymeninx (dura mater) from human spinal cord and brain, expression of mRNA for the beta-trace protein (beta-trace) was studied by in situ hybridization with digoxigenin-labeled cRNA probes. The localization of the protein was investigated using monoclonal and polyclonal antibodies, respectively. Very strong hybridization signals were observed in the fibroblasts of the pachymeninx. The results obtained by in situ hybridization were essentially confirmed by immunocytochemistry. By immunoblotting of proteins solubilized from the dura mater, strong immunoreactions were found with a polyclonal beta-trace antibody. Furthermore, beta-trace was quantified in human ventricular and lumbar cerebrospinal fluid (CSF) by use of an immunonephelometric assay. The beta-trace concentration in human lumbar CSF was elevated 11-fold as compared with ventricular CSF. Beta-trace determined in lumbar CSF most probably originates from fibroblasts of the pachymeninx. According to the bi- or probably multifunctional features of beta-trace, various sites of mRNA expression and protein synthesis exist in the human central nervous system (CNS). The major ones are the fibroblasts of the pachymeninx, and as previously shown, the epithelial layer of the choroid plexus. PMID- 10462697 TI - Sampling of low molecular weight iron by microdialysis following spinal cord injury. AB - We combined the use of desferrioxamine as an iron chelator, microdialysis sampling, and iron analysis by atomic absorption spectroscopy to measure extracellular levels of low molecular weight (LMW) iron in vivo in the spinal cord. Low molecular weight iron is free iron plus iron bound to small molecules. We show that the extracellular LMW-iron concentration is not increased significantly in situ by trauma to the rat spinal cord, suggesting that an extracellular elevation in LMW-iron at the site of injury is not the major initiating factor for lipid peroxidation following spinal cord injury. PMID- 10462698 TI - Structure, chromosomal localization, and brain expression of human Cx36 gene. AB - Rat connexin-36 (Cx36) is the first gap junction protein shown to be expressed predominantly in neuronal cells of the mammalian central nervous system. As a prerequisite for studies devoted to the investigation of the possible role of this connexin in human neurological diseases, we report the cloning and sequencing of the human Cx36 gene, its chromosomal localization, and its pattern of expression in the human brain analyzed by radioactive in situ hybridization. The determination of the human gene sequence revealed that the coding sequence of Cx36 is highly conserved (98% identity at the protein level with the mouse and rat Cx36 and 80% with the ortholog perch and skate Cx35), and that the gene structure is that typical of the Cx35/36 subgroup observed in the other species (presence of a single intron located within the coding region, 71 bp after the translation initiation site). The distribution of Cx36 in several regions of the human central nervous system is similar to that previously observed in rat brain. The most intense signal among the cerebral areas examined by in situ hybridization was observed in the inferior olivary complex, both in principal and accessory nuclei. A moderate labeling was also observed in several myelencephalic nuclei, in specific cells of the the cerebellar cortex, in a relatively large subpopulation of cells in the cerebral cortex, in the hilus of the dentate gyrus, and in the strata radiatum and oriens of hippocampal subfields. Moreover, labeled cells were revealed in all the lamina of the spinal cord gray matter. The chromosomal localization of the human Cx36 gene was determined by fluorescence in situ hybridization. The results allowed assignment of the gene to band 15q14, thus making it a possible candidate gene for a form of familial epilepsy previously linked to the same chromosomal band. The knowledge of the human Cx36 gene sequence, of its chromosomal localization, and of its pattern of expression opens new avenues for the analysis of its possible involvement in human genetic and acquired neuropathology. PMID- 10462699 TI - Inhibition of cyclin-dependent kinases 2 and 4 activities as well as induction of Cdk inhibitors p21 and p27 during growth arrest of human breast carcinoma cells by (-)-epigallocatechin-3-gallate. AB - (-)-Epigallocatechin-3-gallate (EGCG) potently inhibits cell proliferation and suppresses tumor growth both in vitro and vivo, but little is known regarding the cell cycle regulatory proteins mediating these effects. This study investigated the effects of EGCG and other catechins on the cell cycle progression. DNA flow cytometric analysis indicated that 30 microM of EGCG blocked cell cycle progression at G1 phase in asynchronous MCF-7 cells. In addition, cells exposed to 30 microM of EGCG remained in the G1 phase after release from aphidicolin block. Over a 24-h exposure to EGCG, the Rb protein changed from hyper- to hypophosphorylated form and G1 arrest developed. The protein expression of cyclin D1, and E reduced slightly under the same conditions. Immunocomplex kinase experiments showed that EGCG inhibited the activities of cyclin-dependent kinase 2 (Cdk2) and 4 (Cdk4) in a dose-dependent manner in the cell-free system. As the cells were exposed to EGCG (30 microM) over 24 h a gradual loss of both Cdk2 and Cdk4 kinase activities occurred. EGCG also induced the expression of the Cdk inhibitor p21 protein and this effect correlated with the increase in p53 levels. The level of p21 mRNA also increased under the same conditions. In addition, EGCG also increased the expression of the Cdk inhibitor p27 protein within 6 h after EGCG treatment. These results suggest that EGCG either exerts its growth inhibitory effects through modulation of the activities of several key G1 regulatory proteins such as Cdk2 and Cdk4 or mediates the induction of Cdk inhibitor p21 and p27. PMID- 10462700 TI - Distribution of two alternatively spliced variants of the type II collagen N propeptide compared with the C-propeptide in bovine chondrocyte pellet cultures. AB - We have analyzed the distribution of type II collagen N- and C-propeptides in the cell layers and culture medium of bovine articular chondrocyte pellet cultures. Two splice variants of the type II collagen N-propeptide were detected by immunoblotting and immunoassay, using a new anti-peptide antibody, while the C propeptide was detected using a monoclonal antibody. Type II collagen molecules containing the N-propeptide were detected weakly in cell layers, but not in tissue culture medium of chondrocyte pellet cultures, and both splice variants were observed. Free N-propeptide could not be detected in cell layers or medium. Type II procollagen molecules containing the C-propeptide were detected strongly in cell layers, but not in tissue culture medium, while the free C-propeptide was detected in both cell layers and medium. Since the N- and C-propeptides must be synthesized in a 1:1 molar ratio, we conclude that the N-propeptide is metabolized more quickly than the C-propeptide in this system. Our model can be used to study regulation of procollagen synthesis and propeptidase activity. PMID- 10462701 TI - Development of the osteoblast phenotype in primary human osteoblasts in culture: comparison with rat calvarial cells in osteoblast differentiation. AB - In rat osteoblast-like cells, a time-dependent sequence of growth and differentiation-dependent genes has been identified and a model of osteoblast differentiation in culture suggested. We investigated the expression of the bone matrix-associated proteins osteonectin and procollagen I and of the bone cell phenotype-related proteins alkaline phosphatase and osteocalcin during cell culture in primary human osteoblast like cells. Primary human explant cultures from nine young healthy donors were established under highly standardized conditions. Cells in the second passage were analyzed on different days from day 1 to 32, comparing cells growing under the influence of ascorbate with controls. Gene expression was determined by Northern blot analysis or polymerase chain reaction. Osteocalcin expression was also investigated after 1,25-(OH)(2)D(3) stimulation. On the protein level, newly synthesized collagen I, alkaline phosphatase activity, and secretion of osteocalcin were analyzed at all time points. On comparing our findings to the pattern of gene expression suggested for the rat calvarial osteoblast system, we found a similar developmental sequence for the so-called "proliferation" as well as a similar, but lengthened, sequence for the "matrix maturation stage." During "matrix maturation," we found an ongoing proliferation despite increased alkaline phosphatase and decreased procollagen I gene expression. Our study, therefore, shows that in pHOB the gene expression profile proceeded to the "matrix maturation stage," as defined by Owen and colleagues, independent of ongoing proliferation. We were unable to observe the mineralization period as demonstrated by the missing increase of osteocalcin expression and lack of nodule formation in our human osteoblast model. In contrast to the rat system, we found a proliferation stimulating influence of ascorbate, suggesting species-specific differences in response to differentiation factors. From these data, we conclude that general considerations on physiology and pathophysiology of bone cell differentiation have to be confirmed in the human osteoblastic cell system. PMID- 10462702 TI - Spatial distribution of lamin A and B1 in the K562 cell nuclear matrix stabilized with metal ions. AB - When the nucleus is stripped of most DNA, RNA, and soluble proteins, a structure remains that has been referred to as the nuclear matrix, which acts as a framework to determine the higher order of chromatin organization. However, there is always uncertainty as to whether or not the nuclear matrix, isolated in vitro, could really represent a skeleton of the nucleus in vivo. In fact, the only nuclear framework of which the existence is universally accepted is the nuclear lamina, a continuous thin layer that underlies the inner nuclear membrane and is mainly composed of three related proteins: lamins A, B, and C. Nevertheless, a number of recent investigations performed on different cell types have suggested that nuclear lamins are also present within the nucleoplasm and could be important constituents of the nuclear matrix. In most cell types investigated, the nuclear matrix does not spontaneously resist the extraction steps, but must rather be stabilized before the application of extracting agents. In this investigation, by immunochemical and morphological analysis, we studied the effect of stabilization with different divalent cations (Zn(2+), Cu(2+), Cd(2+)) on the distribution of lamin A and B1 in the nuclear matrix obtained from K562 human erythroleukemia cells. In intact cells, antibodies to both lamin A and B1 mainly stained the nuclear periphery, although some immunoreactivity was detected in the nuclear interior. The fluorescent lamin A pattern detected in Cu(2+)- and Cd(2+)-stabilized nuclei was markedly modified, whereas Zn(2+)-incubated nuclei showed an unaltered pattern of lamin A distribution. By contrast, the distribution of lamin B1 in isolated nuclei was not modified by the stabilizing cations. When chromatin was removed by nuclease digestion and extraction with solutions of high ionic strength, a previously masked immunoreactivity for lamin A, but not for lamin B1, became evident in the internal part of the residual structures representing the nuclear matrix. Our results indicate that when metal ions are used as stabilizing agents for the recovery of the nuclear matrix, the distribution of both lamin A and lamin B1 in the final structures, corresponds to the pattern we have very recently reported using different extraction procedures. This observation strengthen the concept that intranuclear lamins may act as structural components of the nuclear matrix. PMID- 10462703 TI - Topoisomerase II is nonfunctional in polyamine-depleted cells. AB - The polyamines-putrescine, spermidine, and spermine-are essential for normal cell proliferation. Polyamine depletion affects DNA structure and synthesis. Topoisomerase II (topo II) is also necessary for normal cell proliferation, and it has been shown in vitro that polyamines may affect topo II activity. In order to investigate the effect of polyamine depletion on topo II activity, we treated Chinese hamster ovary cells with either alpha-difluoromethylornithine (DFMO) or 4 amidinoindan-1-one-2'-amidinohydrazone (CGP 48664), which are polyamine biosynthesis inhibitors. Treatment with the topo II inhibitor etoposide results in DNA strand breaks only if there is active topo II in the cells. By quantitating DNA strand breaks after etoposide treatment using single cell gel electrophoresis, we were able to estimate intracellular topo II activity. We also quantitated topo II activity in crude nuclear extracts from control and polyamine biosynthesis inhibitor-treated cells. Using single cell gel electrophoresis, we noted a clear decrease in the function of topo II in polyamine biosynthesis inhibitor-treated cells, as compared with untreated control cells. However, the topo II activity in crude nuclear extracts did not differ significantly in control versus polyamine biosynthesis inhibitor-treated cells. Taken together, these results indicate that although the function of topo II in polyamine depleted cells was impaired, topo II remained functional in an in vitro assay. Using the single cell gel electrophoresis assay, we also found that spermine depletion itself caused DNA strand breaks. PMID- 10462704 TI - Lack of protein 4.1a in red blood cells of the hereditarily anemic belgrade laboratory (b/b) rat. AB - We have demonstrated that the red blood cell (RBC) membrane of the hereditarily anemic Belgrade laboratory (b/b) rat contains protein 4. 1b isoform, only. The evidence are given that the synthesis of protein 4.1 in the b/b rat reticulocytes is the same as in normal rat. When haemolytic anaemia was induced in normal rat by in vivo phenyhydrazine treatment the same phenomenon, i.e., the absence of protein 4.1a in the RBC membrane was observed. The increase of 4.1a isoform was monitored in RBCs during the recovery of normal rat after phenyhydrazine treatment. Hence, the portion of membrane protein 4.1a isoform is increasing during rat RBC aging. Likewise, when the RBC life span is prolonged (but not normalised) in the b/b rats by iron-dextran treatment protein 4.1a is present in small portion in the RBC membrane. All these data indicate that the lack of protein 4.1a isoform in the b/b rat is due to the presence of young RBCs in the circulation. PMID- 10462705 TI - Colocalization of beta-adrenergic receptors and caveolin within the plasma membrane. AB - The rapid amplification of beta-adrenergic receptor signaling involves the sequential activation of multiple signaling molecules ranging from the receptor to adenylyl cyclase. The prevailing view of the agonist-induced interaction between signaling molecules is based on random collisions between proteins that diffuse freely in the plasma membrane. The recent identification of G protein alpha- and betagamma-subunits in caveolae and their functional interaction with caveolin suggests that caveolae may participate in G protein-coupled signaling. We have investigated the potential interaction of beta-adrenergic receptors with caveolin under resting conditions. beta1- and beta2-adrenergic receptors were recombinantly overexpressed in COS-7 cells. Caveolae were isolated using the detergent-free sucrose gradient centrifugation method. beta1- and beta2 adrenergic receptors were localized in the same gradient fractions as caveolin, where Gsalpha- and betagamma-subunits were detected as well. Immunofluorescence microscopy demonstrated the colocalization of beta-adrenergic receptors with caveolin, indicating a nonrandom distribution of beta-adrenergic receptors in the plasma membrane. Using polyhistidine-tagged recombinant proteins, beta-adrenergic receptors were copurified with caveolin, suggesting that they were physically bound. Our results suggest that, in addition to clathrin-coated pits, caveolae may act as another plasma membrane microdomain to compartmentalize beta adrenergic receptors. PMID- 10462706 TI - Modulation of apo A-IV transcript levels and synthesis by n-3, n-6, and n-9 fatty acids in CACO-2 cells. AB - It has been postulated that apolipoprotein (apo) A-IV plays various significant roles in lipid transport and lipoprotein metabolism. Although it is controlled by fat feeding, so far little else is known about its regulation by specific fatty acids. In this study, we focused on the modulation of apo A-IV mRNA levels, mass, and biogenesis by mono- and polyunsaturated fatty acids (FA) in the human intestinal Caco-2 cell line. In confluent cells incubated with 1 mM oleic (n-9), linoleic (n-6), alpha-linolenic (n-3), or docosahexaenoic (n-3) acids for a long term period, both apo A-IV protein levels and de novo synthesis were increased. The induction resulted from the up-regulation of apo A-IV mRNA transcripts. In contrast, an inhibitory effect was evident with short-term incubation. FA chain length and degree of unsaturation had little effect altering apo A-IV transcript and biogenesis. These data offer evidence that isolated fatty acids regulate gene expression and the production of apo A-IV in the enterocyte. PMID- 10462707 TI - Identification of an autonomous transactivation domain in helix H3 of the vitamin D receptor. AB - The vitamin D receptor (VDR) contains an alpha-helical, ligand-inducible activation function (AF-2) at the COOH-terminus of the ligand-binding domain (LBD). In this study, a second distinct activation domain was identified in the VDR LBD. Using a yeast-based system to screen a random mutant library of GAL4-VDR (93-427), a mutant GAL4-VDR fusion protein with constitutive transcriptional activity was isolated. Sequence analysis identified a C to T transition that introduced a stop codon at glutamine 239 eliminating a large portion of the LBD, including the AF-2 domain. The GAL4-VDR (93-238) mutant exhibited ligand independent transactivation activity both in yeast and in mammalian cells. Deletion analysis defined a minimal activation domain within helix H3 between D195 and I 238 in the VDR. An aspartic acid residue (D232) within helix H3 was essential for the autonomous transactivation activity since altering this residue to an alanine or an asparagine dramatically reduced its transactivation potential. Expression of the minimal helix H3 activation domain interfered with ligand-activated transcription by full-length VDR suggesting that helix H3 interacts with limiting cellular factors important for VDR-activated transcription. Consequently, we have identified a novel activation domain in helix H3 of the VDR that apparently plays an important role in 1,25-(OH)(2)D(3) activated transcription. PMID- 10462708 TI - N-cadherin/catenin-based costameres in cultured chicken cardiomyocytes. AB - N-cadherin is a member of the Ca(2+)-dependent cell adhesion molecules and plays an important role in the assembly of the adherens junction in chicken cardiomyocytes. In addition to being present at the cell-cell junction, N cadherin is associated with costameres in extrajunctional regions. The significance of the N-cadherin-associated costameres and whether catenins are components of costameres in chicken cardiomyocytes are not known. In this study, double-labeling immunofluorescence microscopy was used to determine the extrajunctional distribution of both N-cadherin and its cytoplasmic associated proteins, alpha- and beta-catenins, and their relationship to myofibrillar Z-disc alpha-actinin. N-cadherin, alpha-, and beta-catenins were all found to be present at the extrajunctional region and, in some cases, were codistributed with myofibrillar alpha-actinin exhibiting a periodic staining pattern. Confocal microscopy confirmed that both N-cadherin and beta-catenin colocalized with peripheral myofibrillar alpha-actinin on the dorsal surface of cardiomyocytes as components of the costameres. Intracellular application of antibodies specific for the cytoplasmic portions of N-cadherin, alpha-, and beta-catenin, either by electroporation or microinjection, resulted in myofibril disorganization and disassembly. These results suggest the existence of N-cadherin/catenin-based costameres in the dorsal surface of cultured chicken cardiomyocytes in addition to the integrin/vinculin-based costameres found in the ventral surface and indicate that the former set of costameres is essential for cardiac myofibrillogenesis. PMID- 10462709 TI - Induction of endothelial monolayer permeability by phosphatidate. AB - Released into the vasculature from disrupted cells or transported to the surface of adjacent effectors, phosphatidate and related lipids may potentiate endothelial cell activation. However, the effect of these lipids on endothelial monolayer barrier integrity has not been reported. The present study documents the induction of endothelial monolayer permeability by phosphatidate. Both long (di-C18:1) and medium (di-C10; di-C8) chain length phosphatidates increased permeability of bovine pulmonary artery endothelial cell monolayers assessed using a well characterized assay system in vitro. Barrier disruption effected by dioctanoyl (di-C8) phosphatidate was markedly potentiated by the addition of propranolol, an inhibitor of endothelial cell "ecto"-phosphatidate phosphohydrolase (PAP), a lipid phosphate phosphohydrolase (LPP) that efficiently hydrolyzes extracellular substrate. Disruption of barrier function by phosphatidate did not result from its non-specific detergent characteristics, since a non-hydrolyzable but biologically inactive phosphonate analog of dioctanoyl phosphatidate, which retains the detergent characteristics of phosphatidate, did not induce permeability changes. Furthermore, neither diacylglycerol nor lyso-PA effected significant increases in monolayer permeability, indicating the observed response was due to phosphatidate rather than one of its metabolites. Phosphatidate-induced permeability was attenuated by preincubation of endothelial cells with the tyrosine kinase inhibitor, herbimycin A (10 microg/ml), and enhanced by the tyrosine phosphatase inhibitor, vanadate (100 microM), implicating a role for activation of intracellular tyrosine kinases in the response. In addition, phosphatidate increased the levels of intracellular free Ca(2+) in endothelial cells and ligated specific binding sites on endothelial cell plasma membranes, consistent with the presence of a phosphatidate receptor. Since phosphatidate generated within the plasma membrane of adherent effectors potentially interacts with endothelial membranes, we evaluated the influence of phosphatidate-enriched neutrophil plasma membranes on endothelial monolayer integrity. The effects of ectopic phosphatidate on endothelial monolayer permeability were mimicked by phosphatidate confined to neutrophil plasma membranes. We conclude that phosphatidate may be a physiologic modulator of endothelial monolayer permeability that exerts its effects by activating a receptor-linked, tyrosine kinase-dependent process which results in mobilization of intracellular stored Ca(2+)and consequent metabolic activation. PMID- 10462711 TI - Amino acid availability regulates type I procollagen accumulation in human lung fibroblasts. AB - Fibrotic lung diseases are characterized by excessive deposition of type I collagen. Amino acid availability regulates type I collagen mRNA levels in quiescent human lung fibroblasts. In these studies, the effect of amino acid availability on type I collagen protein accumulation in quiescent human lung fibroblasts was examined. Following amino acid deprivation, alpha1(I) procollagen protein levels were not detected by Western blot analysis in either the intracellular or the extracellular compartments. Fibronectin levels and total protein levels were not affected. Amino acid deprivation resulted in a more pronounced decrease in alpha1(I) procollagen protein levels than in alpha1(I) procollagen mRNA levels, suggesting that post-transcriptional events were responsible for the further decrease inalpha1(I) procollagen protein levels. The addition of transforming growth factor-beta to amino acid deprived fibroblasts increased alpha1(I) procollagen mRNA levels without affecting alpha1(I) procollagen protein levels, confirming a post-transcriptional site for regulatory control by amino acid deprivation. In the absence of ascorbic acid, alpha1(I) procollagen protein levels increased in amino acid deprived fibroblasts, but alpha1(I) procollagen mRNA levels were not affected. The absence of ascorbic acid likely resulted in the accumulation of nonhelical procollagen in the endoplasmic reticulum, indicating that translational mechanisms for alpha1(I) procollagen were intact. The addition of chloroquine, an inhibitor of lysosomal degradation of proteins, increased alpha1(I) procollagen protein levels in amino acid deprived fibroblasts. These data suggest that following amino acid deprivation of quiescent fibroblasts, newly synthesized type I collagen was degraded intracellularly, primarily by a process that involved lysosomal proteinases. PMID- 10462710 TI - Myosin translocation in retinal pericytes during free-radical induced apoptosis. AB - Vascular pathologies induced by ischemia/reperfusion involve the production of reactive oxygen species (ROS) that in part cause tissue injury. The production of ROS that occurs upon reperfusion activates specific second messenger pathways. In diabetic retinopathy there is a characteristic loss of the microvascular pericyte. Pericytes are more sensitive than endothelial cells to low concentrations of ROS, such as hydrogen peroxide (H(2)O(2)) when tested in vitro. Whether the pericyte loss is due to toxic cell death triggered by the noxious H(2)O(2) or apoptosis, due to activation of specific second messenger pathways, is unknown. During apoptosis, a cell's nucleus and cytoplasm condense, the cell becomes fragmented, and ultimately forms apoptotic bodies. It is generally assumed that apoptosis depends on nuclear signaling, but cytoplasmic morphological processes are not well described. We find that exposing cultured retinal pericytes to 100 microM H(2)O(2) for 30 min leads to myosin heavy chain translocation from the cytosol to the cytoskeleton and a significant decrease in cell surface area. Pericyte death follows within 60-120 min. Exposing cells to 150 mJ/cm(2) ultraviolet radiation, an alternate free radical generating system, also causes pericyte myosin translocation and apoptosis. Proteolytic cleavage of actin is not observed in pericyte apoptosis. 3-aminobenzamide, a pharmacological inhibitor of the cleavage and activation of the DNA-repairing enzyme poly (ADP ribose) polymerase (PARP) inhibits pericyte apoptosis, and prevents myosin translocation. Deferoxamine, an iron chelator known to interfere with free radical generation, also inhibits pericyte myosin translocation, contractility, and cell death. Myosin translocation to the cytoskeleton may be an early step in assembly of a competent contractile apparatus, which is involved in apoptotic cell condensation. These results suggest that pericyte loss associated with increased free radical production in diabetic retina may be by an apoptotic phenomenon. PMID- 10462712 TI - Galpha(q/11) and gbetagamma proteins and membrane signaling of calcitriol and estradiol. AB - 17beta-estradiol and 1,25-dihydroxyvitamin D(3)()(calcitriol) rapidly increase (< 5 sec) the concentration of intracellular calcium by mobilizing Ca(2+) from the endoplasmic reticulum and forming inositol 1,4,5-trisphosphate (InsP(3)) and diacylglycerol. Calcitriol increases InsP(3) formation via activation of phospholipase C (PLC)-beta1 linked to a pertussis toxin (PTX)-insensitive G protein, and estradiol via activation of PLC-beta2 linked to a PTX-sensitive G protein. Since PLC are effectors of different subunits of various G-proteins, we looked for and identified several G-subunits (Galpha(q/11), Galphas, Galphai, Gbeta and Ggamma) in female rat osteoblasts using Western immunoblotting. The action of calcitriol on InsP(3) formation and Ca(2+) mobilization in Fura-2 loaded confluent osteoblasts involved Galpha(q/11). The membrane effects of estradiol involved Gbetagamma; subunits, and principally Gbeta subunits, but not alpha-subunits. These results may provide additional evidence for membrane receptors of steroid hormones. Since PLC-beta1 is the target effector of Galpha(q/11), whereas PLC-beta2 is only activated by betagamma subunits, this specificity may help to generate membrane receptor-specific responses in vivo. PMID- 10462713 TI - Flotillin 2 is distinct from epidermal surface antigen (ESA) and is associated with filopodia formation. AB - ECS-1, a monoclonal antibody (MoAb) raised to cultured human keratinocytes, stains the intercellular glycocalyx with a pemphigus-like pattern and recognizes a 35-kDa epidermal surface antigen (ESA) on Western blotting of keratinocyte extracts. When ECS-1 MoAb was used to screen a keratinocyte expression library, a unique cDNA was identified that predicted a 42-kDa globular protein of unknown function. This putative ESA was conserved between mice and humans and was encoded by a gene on chromosome 17q11-12 in linkage with neurofibromin. Homology between the cDNA sequence has been reported with flotillin 1, a caveolae associated protein, as well as Reggie 1 and 2, neuronal proteins expressed during axonal regeneration present in activated GPI-anchored cell adhesion molecules in non caveolar-associated micropatches. In order to determine whether the cDNA predicted protein and ECS-1 antigen were identical, we compared ECS-1 with the immunoreactivity of a new antibody raised to the cDNA fusion protein in epidermis and cultured cells. The cDNA fusion protein was expressed in bacteria and in cos cells with his, FLAG, and EGFP reporter tags and by stable transfection as an EGFP fusion protein. The fusion protein and native protein of 42 kDa were detected by the new antibody, but not by the original ECS-1. Thus, the ECS-1 antigen, ESA (35 kDa), is clearly distinct from the protein predicted by the cDNA (renamed flotillin 2). Stable transfection of ESA/flotillin 2 fusion protein in cos cells induced filopodia formation and changed epithelial cells to a neuronal appearance. Thus, the function of flotillin 2 may resemble that of the goldfish optic nerve neuronal regeneration proteins, Reggie 1 and 2. PMID- 10462714 TI - Characterization of zinc-alpha(2)-glycoprotein as a cell adhesion molecule that inhibits the proliferation of an oral tumor cell line. AB - Zn-alpha(2)-glycoprotein (Znalpha(2)gp) is a soluble protein widely distributed in body fluids and glandular epithelia. We have found it to be expressed in stratified epithelia as well. Znalpha(2)gp is clinically correlated with differentiation in various epithelial tumors, including oral and epidermal tumors. We have cloned epidermal Znalpha(2)gp and report the preparation of the recombinant protein in a Baculovirus expression system. Like the native molecule, recombinant Znalpha(2)gp has RNase activity. Znalpha(2)gp functions as a matrix protein for the Tu-138 oral squamous cell carcinoma cell line. Cell attachment to Znalpha(2)gp is comparable to that for fibronectin and is inhibited by the synthetic RGD peptides RGD, RGDV, and RGDS. Attachment is also inhibited by the antibody to integrin alpha(5)beta(1) (the fibronectin receptor), but not by antibodies to integrins alpha(v)beta(3), alpha(3)beta(1), and alpha(2)beta(1). We find that the proliferation of Tu-138 cells is inhibited on a Znalpha(2)gp matrix, as compared with other matrix proteins (fibronectin, vitronectin, laminin, and collagens I and IV) on which growth resembles that on the BSA control. We believe that the role of Znalpha(2)gp in differentiation and its RNase activity are two likely suspects as agents of the inhibition of proliferation. PMID- 10462715 TI - A magnetic field-responsive domain in the human HSP70 promoter. AB - HSP70 gene expression is induced by a wide range of environmental stimuli, including 60-Hz electromagnetic fields. In an earlier report we showed that the induction of HSP70 gene expression by magnetic fields is effected at the level of transcription and is mediated through c-myc protein binding at two nCTCTn sequences at -230 and -160. in the human HSP70 promoter. We report on the identification of a third c-myc binding site (between -158 and -162) that is an important regulator of magnetic field-induced HSP70 expression. We also show that the heat shock element (HSE), lying between -180 and -203, is required for induction of HSP70 gene expression by magnetic fields. The HSE centered at -100 alone is insufficient. PMID- 10462716 TI - Upregulation of lysyl oxidase in vascular smooth muscle cells by cAMP: role for adenosine receptor activation. AB - Lysyl oxidase (LO) is a key participant in the accumulation of insoluble fibers of elastin and collagen by virtue of its role in the initiation of the covalent cross-linkages between and within individual molecules comprising these fibers. In view of the essential role played by LO in the accumulation of the fibrotic components of occlusive arterial lesions in atherosclerosis, identification of the signaling molecules which can affect the expression of the LO gene in vascular smooth muscle is of considerable interest. In the present report, we describe evidence for the role of the second messenger, cAMP, in the modulation of the levels of LO in vascular smooth muscle cells. Elevated intracellular cAMP induces the transcription of the LO gene, as revealed by Northern blot analysis and nuclear run on assays. Transient transfection experiments performed with the wild-type LO promoter and with this promoter mutated at a consensus CREB site, located within the region -100 to -93 base pairs relative to the start of transcription, indicate that cAMP-induced transcriptional activation is partially due to the presence of this CREB site within the promoter. Activation of stimulatory adenosine receptors in vascular smooth muscle cells with 5'-N ethylcarboxamido adenosine (NECA) increases cAMP, LO mRNA, and enzyme activity. These findings point to the importance of cAMP signaling, potentially initiated by a variety of physiological agents, in the upregulation of LO expression in vascular smooth muscle cells. PMID- 10462717 TI - Spectra of gpt mutations in ethylnitrosourea-treated and untreated transgenic mice. AB - We have established a new transgenic mouse mutagenicity assay for the efficient detection of point mutations and deletions in vivo (Nohmi et al. [1996] Env. Mol. Mutagen. 28:465-470). In this assay, the gpt gene of Escherichia coli is used as a reporter for the detection of point mutations. Treatment of mice with ethylnitrosourea (ENU, 150 mg/kg) enhances by several-fold the mutant frequency of gpt in bone marrow. Here, we report the mutation spectra of the gpt gene recovered from bone marrow of ENU-treated and untreated transgenic mice. In the gpt mutants rescued from ENU-treated mice, more than 90% of the mutations were base change mutations; the predominant types were A:T to T:A transversions and G:C to A:T transitions. On the contrary, in the mutants rescued from untreated mice, 54% were base substitutions and the remainders were short deletions and insertions. Among untreated mice, the most frequently observed base substitution was G:C to A:T transitions (7/14 mutants). Three of these occurred at 5'-CpG-3' sites. Interestingly, the mutation spectra of the gpt gene were different from those of the gpt gene in ENU-treated and untreated E.coli, whereas they were similar to those of the lacZ and lacI genes in ENU-treated and untreated other transgenic mice or cultured mammalian cells. We also report the establishment of homozygous transgenic mice that have transgene lambdaEG10 DNA in both chromosome 17 of C57BL/6J mouse. PMID- 10462718 TI - Spi(-) selection: An efficient method to detect gamma-ray-induced deletions in transgenic mice. AB - Despite the importance of genome rearrangement in the etiology of cancer and human genetic disease, deletion mutations are poorly detectable by transgenic rodent mutagenicity tests. To facilitate the detection and molecular analysis of deletion mutations in vivo, we established a transgenic mouse model harboring a lambdaEG10 shuttle vector that includes the red and gam genes for Spi(-) (sensitive to P2 interference) selection [Nohmi et al. (1996] Environ. Mol. Mutagen. 28:465-470]. This selection has a great advantage over other genetic systems, because phage deletion mutants can be preferentially selected as Spi(-) plaques, which can then be subjected to molecular analysis. Here, we show nucleotide sequences of 41 junctions of deletion mutations induced by gamma irradiation. Unlike spontaneous deletion mutants, more than half of the large deletions occurred between short homologous sequences from one to eight bp. The remaining junctions had no such homologous sequences. Intriguingly, two Spi(-) mutants had P (palindrome)-like nucleotide additions at the breakpoints, which are frequently observed in the coding junctions of V(D)J recombination, suggesting that broken DNA molecules with hairpin structures can be intermediates in the repair of radiation-induced double-strand breaks. We conclude that Spi(-) selection is useful for the efficient detection of deletion mutations in vivo and that most rearrangements induced by gamma-rays in mice are mediated by illegitimate recombination through DNA end-joining. PMID- 10462719 TI - Significance of the perigametic interval as a major source of spontaneous mutations that result in mosaics. AB - An earlier analysis showed that a significant percentage of spontaneous specific locus mutations in mice are recovered as mosaics and that the spontaneous mutation rate per cell cycle is probably higher for those mutations that produce mosaics than for those that produce whole-body mutants. The finding that the average germline composition of the mosaics was approximately 50% supported the suggestion that single-strand DNA alterations during the perigametic interval constitute the major source of spontaneous mosaics. Here, alternative origins of 50% germline mosaicism are examined. Supporting the earlier hypothesis is the finding that spontaneous mutations that are recovered as clusters constitute a different array of types from those giving rise to singletons, and the evidence from interspecies comparisons that a unique component of the life cycle, probably meiosis, makes a major contribution to spontaneous mutations. Biological factors associated with the perigametic interval were examined in an effort to suggest explanations for the observations that 1) the spontaneous mutation rate in that interval is high relative to that characterizing any mitotic cell cycle, 2) the types of mutations appear to be different from those arising during mitotic divisions, and 3) the spontaneous mutation rate for males is higher than that for females. It is concluded that the higher yield from the perigametic interval is consistent with what is known about methylation status in development of both sexes and with repair capacity in the male germline. For both parameters, differences between the sexes during their respective perigametic intervals may be at least partly responsible for the fact that the spontaneous mutation rate of mammalian females is lower than that of males. PMID- 10462720 TI - Induced mutations in M13mp2 phage DNA exposed to N-nitrosopyrrolidine with UVA irradiation. AB - It is known that N-nitrosopyrrolidine (NPYR), a carcinogen in rodents, is metabolically activated by microsomal cytochrome P450 to form an alpha hydroxylated derivative, which induces mutations. The mutations have been demonstrated by use of Salmonella typhimurium and Escherichia coli. We discovered directly acting mutagenicity of NPYR plus ultraviolet light-A (UVA) in bacteria and phage. With an O(6)-alkyltransferase-deficient strain of S.typhimurium, the NPYR plus UVA treatment gave greater mutation frequencies compared to those found with the parent strain. We identified the structure of the direct-acting mutagen as N-nitroso-1-phosphonooxypyrrolidine, and analyzed the spectrum of mutations induced in the DNA of M13mp2 phage. The basepair substitutions GC to TA and GC to AT appear to occur predominantly. Several hotspots were observed. In the conditions where SOS response was induced in the host E.coli, greater varieties of mutations were observed in phage DNA compared to those without the SOS response induction. These results suggest that alkylations of DNA occur by the photoactivated NPYR. The roles of the produced damage to the mutations are discussed. PMID- 10462721 TI - Molecular analysis of in vivo mutations induced by N-ethyl-N-nitrosourea in the autosomal Tk and the X-linked Hprt genes of mouse lymphocytes. AB - The endogenous, autosomal Tk gene is a potentially useful reporter of in vivo mutation since it may recover a wider range of mutational events than the X linked Hprt gene or bacterial transgenes. In this study, we characterized mutations produced in the Tk gene of Tk(+/-) mice and compared them with mutations induced in the Hprt gene. Treatment of Tk(+/-) mice with N-ethyl-N nitrosourea (ENU) resulted in dose-related increases in Tk mutants, as measured by the frequency of 5-bromodeoxyuridine-resistant (BrdUrd(r)) spleen lymphocytes. ENU-induced mutant frequencies in the Hprt gene, determined by measuring 6 thioguanine-resistant (TG(r)) lymphocytes, were similar to the Tk mutant frequencies. Allele-specific PCR of DNA from BrdUrd(r) lymphocyte clones suggested that 35% of clones from mice treated with ENU and 65% of clones from untreated animals had loss of heterozygosity (LOH) of the Tk gene due to deletion of the functional Tk allele. Reverse transcriptase-PCR/sequencing analysis of BrdUrd(r) and TG(r) clones from ENU-treated mice indicated that point mutations in both genes predominantly occurred at A:T basepairs; however, A:T-->G:C transition was the most common mutation in the Tk gene, while A:T-->T:A transversion was the most frequent mutation in the Hprt gene. Substitution at A:T basepairs in the Hprt gene occurred disproportionately with the mutated dT on the nontranscribed DNA strand, while this strand bias for mutation was not seen in the Tk gene. The results indicate that the specificity of ENU-induced point mutation differs between the two endogenous genes and that the autosomal Tk gene of Tk(+/-) mice is capable of recovering mutations caused by LOH. Environ. Mol. Mutagen. 34:30-38, 1999 Published 1999 Wiley-Liss, Inc. PMID- 10462722 TI - Aneuploidy in sperm and exposure to fungicides and lifestyle factors. ASCLEPIOS. A European Concerted Action on Occupational Hazards to Male Reproductive Capability. AB - Fungicides include chemicals that are known aneugens. The purpose of the present study was to investigate whether occupational exposure to these and other agricultural pesticides induces aneuploidy in human sperm. The contribution of lifestyle factors (smoking and alcohol consumption) to the frequency of aneuploid sperm was evaluated as well. The effects of age and sperm concentration were analyzed as confounders. Spermatozoa from 30 healthy farmers were studied before and after exposure to fungicides, using fluorescence in situ hybridization (FISH). Ten thousand spermatozoa were scored per semen sample to determine the disomy and diploidy frequencies for chromosomes 1 and 7. Exposure to fungicides was not associated with sperm aneuploidy. Smoking was significantly associated with sperm carrying an extra chromosome 1 and with diploid sperm as well as with the aggregate frequency of aneuploid sperm. Alcohol consumption, sperm concentration, and age showed inconsistent results before and after the season of exposure to fungicides. For low-level exposures, such as occupational exposures, the sensitivity of the sperm-FISH method may not be sufficient. The present study supports earlier ones showing that smoking can increase aneuploidy in human sperm. PMID- 10462723 TI - Genotoxic activity of different chromium compounds in larval cells of Drosophila melanogaster, as measured in the wing spot test. AB - Two chromium(VI) compounds (potassium chromate and potassium dichromate) and one chromium(III) compound, chromium chloride, were evaluated for genotoxic effects in the wing spot test of Drosophila melanogaster following standard procedures. This assay detects both somatic recombination and mutational events. The genotoxic effects were determined from the appearance of wing spots in flies transheterozygous for the third chromosome recessive markers multiple wing hairs (mwh) and flare-3 (flr(3)), as well as in flies heterozygous formwh and the multiply inverted TM3 balancer chromosome. Genetic changes induced in somatic cells of the wing's imaginal discs lead to the formation of mutant clones on the wingblade. Single spots are due to different genotoxic mechanisms: point mutation, deletion, chromosome breakage, and mitotic recombination; while twin spots are produced only by mitotic recombination. From our results it appears that both chromium(VI) compounds clearly increase the incidence of mutant clones by inducing high increases in the frequency of all types of clones recorded. On the contrary, chromium(III) did not increase the frequency of mutant clones. A high proportion of the total spot induction was due to mitotic recombination, confirming previously reported data on the strong recombinogenic activity of chromium(VI) compounds. PMID- 10462724 TI - Exposure to agrochemicals and DNA adducts in Western Liguria, Italy. AB - Pesticides are used to control pests and improve agricultural production. Despite their selectivity of action, a number of agrochemicals have been reported to be genotoxic using the (32)P-DNA postlabeling assay. Greenhouse floriculturists are suspected of being heavily exposed to agrochemicals during loading, mixing, and application of pesticides, as well as during manual activities by continuous contact with flowers and ornamental plants. We analyzed the DNA adduct formations in the white blood cells (WBCs) of 57 nonsmoker greenhouse floriculturists and 33 nonsmoker age-matched referents residing in the Western Liguria Region, Italy-the most important Italian greenhouse floriculture area. The averages of DNA adducts, expressed as relative adduct labeling (RAL), were 8.50 x 10(9) +/- 1.98 (SE) in floriculturists and 2.17 x 10(9) +/- 1.05 (SE) in referents. DNA adducts were significantly higher in floriculturists than in controls after adjustment for age and gender (P = 0.007). A specific adduct pattern, with up to six different spots, was observed in 60% of floriculturists, while no adducts were generally detected in controls. Our study represents an important contribution to the correct evaluation of the potential health risk associated with floriculture activity and supports the adoption of measures ensuring pesticide exposure reduction in greenhouses. PMID- 10462725 TI - Inhibitory effects of chlorophyllin on micronucleus formation induced by ethyl carbamate and its proximate and ultimate carcinogenic forms in mouse peripheral reticulocytes. PMID- 10462726 TI - Meiotic nondisjunction induced by furfural in Drosophila melanogaster females. PMID- 10462727 TI - Detection of chromosome loss and gain induced by griseofulvin, estramustine, and vanadate in binucleated lymphocytes using FISH analysis. AB - Cytochalasin B-blocked binucleated human lymphocytes from a healthy male donor were used to detect micronucleus induction and other aneuploidy events (chromosome loss and gain) after treatment with griseofulvin (GF), estramustine (EM), and sodium orthovanadate (Na(3)VO(4)). A two-color FISH was performed by using centromeric probes for chromosome 2 (FITC labeled) and the X chromosome (TRITC labeled) to measure chromosome loss and gain events in binucleated cells. GF induced mainly aneuploid binucleates involving the X chromosome, but this was not associated with preferential loss of one of the two chromosomes. EM preferentially induced aneuploidy of chromosome 2, and Na(3)VO(4) of the X chromosome. Our results indicate that chromosome malsegregation events (chromosome loss and/or gain) are probably not randomly induced, suggesting that different mechanisms leading to aneuploidy may be either chromosome-dependent or compound- and dose- related. PMID- 10462728 TI - Clinical anatomy of internal thoracic artery branches. AB - The broad clinical utilization of the internal thoracic artery (ITA), including the role of its branches in supplying circulation to the sternum, requires explicit anatomic knowledge of this vessel. Fifty-six ITAs (28 right, 28 left) were dissected from their point of origins after injection with a mixture of contrast medium and latex after perfusion with saline and immersion in 4% formaldehyde. All ITA branches were studied according to their course, size, and distribution within intercostal spaces with the aid of an operating microscope. The branches were divided in two main groups: proper (solitary) branches and common trunks. The proper branches consisted of four types: sternal, perforating, intercostal, and mediastinal. The four types of common trunks were: sternal/perforating, sternal/intercostal, perforating/intercostal, sternal/perforating/intercostal. Points of most frequent origin from main trunk of the vessel were established for each type. Mean external diameter of proper branches was 0.72 mm and common trunks was 1.06 mm. Mean length of common trunks was 3.0 mm. Those parameters (adequate diameter and length) allow for ligation of the common trunks close to the ITA so that their points of division can be preserved. This fact is crucial for creation of collateral blood supply to the sternum after bilateral ITA mobilizations. PMID- 10462730 TI - CT imaging and three-dimensional reconstructions of shoulders with anterior glenohumeral instability. AB - Glenohumeral instability is a common occurrence following anterior dislocation of the shoulder joint, particularly in young men. The bony abnormalities encountered in patients with glenohumeral instability can be difficult to detect with conventional radiography, even with special views. The aim of our study was to evaluate the bony abnormalities associated with glenohumeral instability using CT imaging with 3-D reconstruction images. We scanned 11 patients with glenohumeral instability, one with bilateral symptoms; 10 were male, one female, and their ages ranged from 18-66 years. Contiguous 3 mm axial slices of the glenohumeral joint were taken at 2 mm intervals using a Siemens Somatom CT scanner. In the 12 shoulders imaged, we identified four main abnormalities. A humeral-head defect or Hill-Sachs deformity was seen in 83% cases, fractures of the anterior glenoid rim in 50%, periosteal new bone formation secondary to capsular stripping in 42%, and loose bone fragments in 25%. Manipulation of the 3-D images enabled the abnormalities to be well seen in all cases, giving a graphic visualization of the joint, and only two 3-D images were needed to demonstrate all the necessary information. We feel that CT is the imaging modality most likely to show all the bone abnormalities associated with glenohumeral instability. These bony changes may lead to the correct inference of soft tissue abnormalities making more invasive examinations such as arthrography unnecessary. PMID- 10462729 TI - Spatial and temporal distribution of cellular proliferation in the cranial base of normal and midfacially retrusive mice. AB - The craniofacial region of the Brachyrrhine (Br) mouse is characterized by a retruded midface. The cellular mechanism causing this growth deficiency is unknown. However, the cranial base is foreshortened in adult Br mice. The purpose of this study was to determine whether the spatial and temporal patterns of cellular proliferation in the cranial base (CB) differ between normal (C3H/HeJ) and Br mutant (3H1 Br/+) embryonic mice. Twenty-four dams were injected (3)H thymidine (5 microCi/gram body weight) and 15 embryos from each group were collected at Theiler stages 23, 25, and 27 (15, 17, and 19 days of gestation). Serial sections from each head were processed with routine autoradiography. Labelling indices (LI) were determined for each specimen and cellular proliferation maps were generated for each age group. LI patterns within and between groups were compared statistically. Results showed that cellular proliferation in the CB of normal embryos displayed a time- and position dependent pattern, characteristic of transient growth sites (TGS). Generally, as age increases, cellular proliferative activities decrease gradually (from an average LI of 11.4 +/- 5.7% at stage 23 to 4.4 +/- 2.2% at stage 27), and the number of the TGS decreases in the presumptive nasal septal region and increases in presumptive sphenoethmoidal area with age, indicating the existence of cellular subpopulations in the CB. Cellular proliferation in the CB of the Br mutant displays a different growth pattern compared to the normal condition. Deficiencies in cellular proliferation exist mainly in the presumptive sphenoethmoidal area of the CB. The results indicate that the TGS play an important role in the normal morphogenesis of the CB, and abnormalities in their timing and/or position may be responsible for the dysmorphology of the midface in the Br mutant. PMID- 10462731 TI - Handouts as an educational support for the teaching/learning program in clinical anatomy. AB - Handouts were developed to support the program of Clinical Anatomy in the Medical School of Porto, and since 1996/97 alterations have been made to improve their format and content with our educational objectives in mind. A questionnaire was designed to evaluate the opinion of second-year medical students enrolled in the program. Students were asked about their approval of the way handouts were organized and their usefulness, especially for lectures and practical sessions on physical examination, sectional and imaging anatomy, anatomical variations and malformations and case studies. Of 152 students, 138 (90.8%) returned the questionnaire. To describe the relationship between the value of handouts and several aspects of their organization and adequacy, the Spearman rank correlation coefficient was used for lectures, and canonical correlation for the various practical sessions. Students fully approved the way the handouts of lectures and practical sessions were organized (81.8% and 87%, respectively), their presentation (74.6% and 86.2%), relevance (88.3% and 85.5%), usefulness in understanding the lectures (77.6%) and their value in preparing for practical sessions (83.3%). Handouts were considered highly useful for case studies (90%), physical examination (81.9%) and sectional anatomy (65.7%). Students stating a higher degree of utility of the handouts emphasized that they were useful-indeed essential-in preparing for sessions, and noted their objectivity. The evaluation of the handouts was highly favorable and showed that they can be used as a guide through the complexities of an innovative program of Clinical Anatomy. PMID- 10462732 TI - Horner's syndrome revisited: with an update of the central pathway. AB - A brief summary is presented of the life of Johann Friedrich Horner, the eminent Swiss ophthalmologist, renowned for describing the effects of paralysis of the human cervical sympathetic nerves. His early education, the quality of his professional training, and the influence of his mentors, notably Carl Ludwig and Albrecht von Graefe, contributed to his discovery of the syndrome. The full text of Horner's original work (translated by J. F. Fulton, 1929a, Arch. Surg. 18:2025 2039) is cited. The history of clinical and experimental work carried out on the autonomic nervous system prior to Horner's discovery is reviewed, including the studies of Pourfour du Petit (cited in Fulton, 1929a and Singer and Underwood, 1962, Clarendon); Hare, 1838, Lond. Med. Gaz. 23:16-18; Bernard (cited by Singer and Underwood); Budge (1853, Acad. de Sci., p.377-378); Mitchell et al. (1864, Lippincott). Hare and Mitchell et al. came close to making the discovery but were apparently hindered by their inability to interpret the signs they elicited in their patients. The experiments of Claude Bernard gave succinct accounts of the effects of damage to the cervical sympathetic nerves in animals, although there appears to be no evidence that he made similar observations in humans. Horner was the first to give a detailed, scientifically supported account and accurately interpret the signs of cervical sympathetic nerve damage in a human subject. The anatomy of the pathway is reviewed and the detailed structure of its central part updated. Evidence from computerized tomography (CT), magnetic resonance imaging (MRI), positron emission tomography (PET), and single-photon-emission computerized tomography (SPECT) studies have confirmed that reciprocally connected centers in the insular cortex, central nucleus of amygdala, hypothalamus, mesencephalic and pontine tegmentum, nucleus of tractus solitarius, and the ventrolateral medulla form the central pathway. The nucleus of tractus solitarius is probably the main reflex center for the sympathetic system, whereas the ventrolateral medulla serves as the pathway through which the central neurons influence the preganglionic neurons of the thoracolumbar outflow. Emotional and sensory inputs from the frontal and somatosensory cortices provide the inputs needed by the insula to drive the sympathetic nervous system to produce appropriate responses. PMID- 10462733 TI - Clinical anatomy of the atrial septum with reference to its developmental components. AB - Knowledge of development is of crucial importance and can help clarify mechanisms of maldevelopment, but it must be properly validated. Concepts of development must be consistent with the anatomy seen in postnatal life. Such consistency is not always achieved. We have reviewed new and old accounts of cardiac embryology with regard to the definitive structure of the atrial septum. The key to understanding is to distinguish between folds of the atrial wall and true interatrial partitions. The flap valve of the oval foramen, and its inferior rim, are true septal structures, whereas the other rims, particularly the antero superior rim, are infoldings enclosing extracardiac fat. During embryonic life, the systemic venous tributaries must achieve entrance only to the right side of the primary atrium. Development of the pulmonary venous component is a late event, with the canalizing vein using the dorsal mesocardium to gain access to the left side of the atrium. Once the systemic venous tributaries have achieved their rightward shift, the primary septum, together with the mesenchymal cap, grows between the systemic and pulmonary venous orifices. Closure of the primary foramen is achieved by fusion of the mesenchymal cap of the primary septum with the atrioventricular endocardial cushions and the vestibular spine (an additional mesenchymal structure carried on the right side of the pulmonary venous orifice). The superior margin of the newly formed secondary foramen is produced by an infolding of the atrial walls. Historically these mechanisms received appropriate recognition, but not all receive their proper due in current writings. PMID- 10462734 TI - The spinal epidural space. AB - The validity of the concept of an epidural 'space' within the vertebral canal is questioned. An attempt is made to locate the 'space' morphologically, developmentally, and topographically. Following Parkin and Harrison (1985), it is agreed that no actual 'space' exists in the intact living subject. It is suggested that the spinal epidural space satisfies the criteria of Haines (1991) to be considered a "true potential space." As such, and by analogy with other body cavities, it would be better termed the 'epidural cavity', as it was in the 1983 edition of the Nomina Anatomica. PMID- 10462735 TI - Teaching of anatomy in the Philippine setting. PMID- 10462736 TI - Morphologic determinants in the etiology of class III malocclusions: a review. AB - Morphospatial disharmony of the craniomaxillary and mandibular complexes may yield apparent mandibular prognathism, but Class III malocclusions can exist with any number of aberrations of the craniofacial complex. Deficient orthocephalization of the cranial base allied with a smaller anterior cranial base component has been implicated in the etiology of Class III malocclusions. Whereas the more acute cranial base angle may affect the articulation of the condyles resulting in their forward displacement, the reduction in anterior cranial size may affect the position of the maxilla. As well, intrinsic skeletal elements of the maxillary complex may be responsible for maxillary hypoplasia that may exacerbate the anterior crossbite seen in the Class III condition. Conversely, with an orthognathic maxilla, condylar hyperplasia and anterior positioning of the condyles at the temporo-mandibular joint may produce an anterior crossbite. Aside from the skeletal components, soft tissue matrices, particularly labial pressure from the circumoral musculature, may influence the final outcome of craniofacial growth of a child skeletally predisposed to Class III conditions. Indeed, as some Asian ethnic groups demonstrate an increased prevalence of Class III malocclusions, it is likely that the skeletal components and soft tissues matrices are genetically determined. Presumably, the co morphologies of the craniomaxillary and mandibular complexes are likely dependent upon candidate genes that undergo gene-environmental interactions to yield Class III malocclusions. The identification of such genes is a desirable step in unraveling the complexity of Class III malocclusions. With this knowledge, the clinician may elect an early course of dentofacial orthopedic and orthodontic treatments aimed at preventing the development of Class III malocclusions. PMID- 10462737 TI - Immunity and probiotics. PMID- 10462738 TI - Granulysin: a lethal weapon of cytolytic T cells. PMID- 10462739 TI - Allergy to lipocalins: a consequence of misguided T-cell recognition of self and nonself? PMID- 10462740 TI - Positive and negative regulation of Src-family membrane kinases by CD45. PMID- 10462741 TI - CD45 and Src-family kinases: and now for something completely different. PMID- 10462742 TI - Binding energetics of T-cell receptors: correlation with immunological consequences. PMID- 10462743 TI - Critical self-epitopes are key to the understanding of self-tolerance and autoimmunity. PMID- 10462744 TI - Thymic involution and HIV progression. PMID- 10462745 TI - Germline gene transfer during gene therapy: reassessing the risks. PMID- 10462746 TI - Aggregation inhibitors: hope for Huntington's? PMID- 10462747 TI - Nanobacteria linked to kidney disease. PMID- 10462748 TI - On the TRAIL to a new cancer therapy. PMID- 10462749 TI - Does paraoxonase play a role in susceptibility to cardiovascular disease? AB - Human serum paraoxonase (PON1) is an esterase that is bound to high-density lipoproteins (HDLs). It can hydrolyze organophosphates and its activity is inversely related to atherosclerosis. Some studies also suggest that a relationship exists between polymorphisms of the gene that encodes paraoxonase and coronary heart disease (CHD), whereas other studies, in different populations, have not found such an association. One mechanism by which certain PON1 allozymes might protect against atherosclerosis is by inhibition of the oxidation of HDL and low-density lipoprotein (LDL). Experimental studies suggest that this protection is associated with the ability of PON1 to hydrolyze specific lipid peroxides in oxidized lipoproteins. Interventions that preserve or enhance PON1 activity, as well as manipulations of PON1 polymorphisms, might help delay the onset of CHD. PMID- 10462750 TI - The p53 family: same response, different signals? AB - TP53, the gene that encodes p53, is a well-defined tumor suppressor gene that is frequently mutated in human cancers. Recently, two proteins homologous to p53, termed p73 and p63, were identified. Current data indicate that both p73 and p63, like p53, can induce cell-cycle arrest and apoptosis, suggesting that they might also be tumor suppressors. However, the physiological signals that can regulate p53, for example, DNA damage, have no effect on p73, as tested in several cell lines. Furthermore, the signaling pathways by which p73 (and possibly p63) induces cell-cycle arrest and apoptosis appear to be similar to those of p53, but also have important differences. Thus, the p53 family proteins are closely related but might have distinct physiological functions. PMID- 10462751 TI - Hepatitis C virus: current understanding and prospects for future therapies. AB - Hepatitis C virus (HCV) is the leading cause of chronic liver disease worldwide and the leading indication for liver transplantation. The hallmark of the disease is its propensity to evolve into chronicity, probably because viral heterogeneity allows the virus to escape immune-mediated neutralization. Treatment with interferon alpha (IFN-alpha) has been disappointing, but higher and more frequent doses, and combination therapies, including nucleoside analogs, might lead to improved suppression of HCV RNA levels. Molecular analysis of HCV before and during treatment has indicated that high viral RNA levels and the presence of HCV genotype 1 are independent predictors of poor treatment outcome. New antiviral agents in development include inhibitors of HCV replicative enzymes, such as protease, helicase and polymerase, as well as several genetic approaches, such as ribozymes and antisense oligonucleotides. The main hindrance to drug development for hepatitis C is the lack of a small animal model or a productive tissue culture system for assessing drug action. PMID- 10462752 TI - Gene therapy strategies to facilitate organ transplantation. AB - Organ transplantation is now the definitive therapy for many forms of end-organ disease, but chronic allograft rejection, the side effects of chronic immunosuppressive therapy and the severe donor organ shortage continue to limit its success. Gene therapy has the potential to prevent graft rejection by manipulating the immune response in the microenvironment of the graft or by facilitating the induction of tolerance. Genetic manipulation of stem cells to create transgenic and/or knockout animals that could serve as organ or cell donors could be combined with gene therapy approaches to overcome the problem of limited allogeneic donor organ supply. PMID- 10462753 TI - The oncostatin M signalling pathway: reversing the neoplastic phenotype? AB - Oncostatin M (OSM) is a member of the interleukin 6 (IL-6) family of cytokines and was originally identified by its ability to inhibit proliferation of melanoma cells but augment the growth of normal fibroblasts. OSM has pleiotropic effects on many different cell types, but here we focus on its ability to inhibit the proliferation of cell lines derived from several tumour types, including breast carcinoma, ovarian cancer, melanoma, glioma and lung carcinoma. The inhibition of proliferation of several cancer cell lines by OSM is associated with alterations in cellular morphology and with phenotypic changes that are consistent with the induction of differentiation of these cells. These observations raise the possibility that OSM could have therapeutic potential. PMID- 10462754 TI - An animal model for pycnodysostosis: the role of cathepsin K in bone remodelling. PMID- 10462756 TI - Receptor dimerization and Hill coefficients. PMID- 10462755 TI - The race to control pain: more participants, more targets. PMID- 10462757 TI - Onaran and Gurdal reply PMID- 10462758 TI - Metalloporphyrin class of therapeutic catalytic antioxidants. AB - Metalloporphyrins have emerged as a novel class of catalytic antioxidants that scavenge a wide range of reactive oxygen species (ROS) such as superoxide, peroxide, peroxynitrite and lipid peroxyl radicals. Factors such as the type of metal centre, redox potential and electrostatic charge of the compounds are recognized as important determinants of their antioxidant activity and potency. These concepts have guided the development of metalloporphyrins with specific activities greater than those of the native superoxide dismutases. Several compounds in this class have been shown to be efficacious in a variety of in vitro and in vivo oxidative stress models of human diseases. PMID- 10462759 TI - Potential drug targets: small GTPases that regulate leukocyte function. AB - Leukocytes are not only important mediators of innate immunity, but they also induce and perpetuate inflammatory responses that are harmful to the host. Although inflammatory mediators activate leukocytes through a common heterotrimeric G protein (Gi) signalling intermediate, many downstream inflammatory functions are regulated by distinct small GTPases, which suggests that pharmacological modulation of small GTPase activity would be useful in developing specific anti-inflammatory therapies. The recent identification of multiple small GTPase effectors, the recognition of the role of GTPase regulatory proteins in directing downstream signalling from small GTPases, and detailed structural information on the GTPases themselves suggests new possibilities for the development of effective and selective anti-inflammatory drugs. PMID- 10462760 TI - Novel GPCRs and their endogenous ligands: expanding the boundaries of physiology and pharmacology. AB - Nearly all molecules known to signal cells via G proteins have been assigned a cloned G-protein-coupled-receptor (GPCR) gene. This has been the result of a decade-long genetic search that has also identified some receptors for which ligands are unknown; these receptors are described as orphans (oGPCRs). More than 80 of these novel receptor systems have been identified and the emphasis has shifted to searching for novel signalling molecules. Thus, multiple neurotransmitter systems have eluded pharmacological detection by conventional means and the tremendous physiological implications and potential for these novel systems as targets for drug discovery remains unexploited. The discovery of all the GPCR genes in the genome and the identification of the unsolved receptor transmitter systems, by determining the endogenous ligands, represents one of the most important tasks in modern pharmacology. PMID- 10462761 TI - Emerging roles for RGS proteins in cell signalling. AB - Regulators of G-protein signalling (RGS proteins) are a family of highly diverse, multifunctional signalling proteins that share a conserved 120 amino acid domain (RGS domain). RGS domains bind directly to activated Galpha subunits and act as GTPase-activating proteins (GAPs) to attenuate and/or modulate hormone and neurotransmitter receptor-initiated signalling by both Galpha-GTP and Gbetagamma. Apart from this structural domain, which is shared by all known RGS proteins, these proteins differ widely in their overall size and amino acid identity and possess a remarkable variety of structural domains and motifs. These biochemical features impart signalling functions and/or enable RGS proteins to interact with a growing list of unexpected protein-binding partners with diverse cellular roles. New appreciation for the broader cellular functions of RGS proteins challenges established models of G-protein signalling and serves to identify these proteins as central participants in receptor signalling and cell physiology. PMID- 10462763 TI - A family of phytochelatin synthase genes from plant, fungal and animal species. PMID- 10462762 TI - GPCR-Galpha fusion proteins: molecular analysis of receptor-G-protein coupling. AB - The efficiency of interactions between G-protein-coupled receptors (GPCRs) and heterotrimeric guanine nucleotide-binding proteins (G proteins) is greatly influenced by the absolute and relative densities of these proteins in the plasma membrane. The study of these interactions has been facilitated by the use of GPCR Galpha fusion proteins, which are formed by the fusion of GPCR to Galpha. These fusion proteins ensure a defined 1:1 stoichiometry of GPCR to Galpha and force the physical proximity of the signalling partners. Thus, fusion of GPCR to Galpha enhances coupling efficiency can be used to study aspects of receptor-G-protein coupling that could not otherwise be examined by co-expressing GPCRs and G proteins as separate proteins. The results of studies that have made use of GPCR Galpha fusion proteins will be discussed in this article, along with the strengths and limitations of this approach. PMID- 10462764 TI - From gene shuffling to the restoration of riparian ecosystems. PMID- 10462765 TI - Gene flow and GM questions. PMID- 10462766 TI - Listening to the silent genes: transgene silencing, gene regulation and pathogen control. AB - By capitalizing on the initially puzzling observations of unpredictable transgene silencing and variable expression, plant scientists have pioneered research into a novel type of epigenetic regulation, termed homology-dependent gene silencing. This silencing process has implications for natural mechanisms of gene expression in plants and other eukaryotes, and has branched out into studies of reversible DNA modifications; RNA metabolism, transport and processing; and host responses to plant viruses, viroids and transposable elements. The analysis of transgene silencing systems has enriched our understanding of other epigenetic phenomena, including paramutation, as well as heterosis and genome evolution. This research is also highly relevant to the biotechnology industry, which is interested in avoiding unwanted transgene silencing in genetically engineered lines and in exploiting various types of silencing to inactivate specific genes. Homology dependent gene silencing can also be used in high-throughput approaches for functional genomics. PMID- 10462767 TI - Brassinosteroid biosynthesis inhibitors. AB - Recent work on the physiological responses of brassinosteroid-deficient mutants has led to the designation of brassinosteroids as a new class of phytohormone. However, information on other possible roles of brassinosteroids is limited because the mutant analysis has been confined to a relatively small number of plant species. In this context, specific inhibitors of brassinosteroid biosynthesis would be valuable for investigating the roles of brassinosteroids at various stages of plant development, such as germination, leaf expansion and flowering. Data on the oxidative processes involved in brassinosteroid biosynthesis have permitted the rational design of biosynthesis inhibitors. The development of such inhibitors has confirmed the importance of brassinosteroids in broad aspects of plant growth and development, with drastic morphological changes in inhibitor-treated plants, almost identical to those found in brassinosteroid-deficient mutants. The normal phenotype of inhibitor-treated plants can be recovered by the addition of brassinolide, revealing that inhibitors can be used to complement brassinosteroid-deficient mutants in clarifying the functions of brassinosteroids. PMID- 10462768 TI - Macromolecular trafficking in the phloem. AB - Prior to the early 1990s, the functional aspects of long-distance movement in the phloem were viewed primarily in terms of the transport of sugars and other photoassimilates. The soluble proteins in phloem exudates were often considered to be artifacts arising from the cytoplasmic degeneration of the conducting elements or a confounding anomaly of the sugar transport system. Recent work indicates a much more complex system where proteins and other macromolecules play significant roles in phloem function and long-distance signaling throughout the plant. PMID- 10462769 TI - Xyloglucan endotransglycosylases: diversity of genes, enzymes and potential wall modifying functions. AB - Plant cells are enclosed by walls that define the shapes and sizes of cells and mediate cell-to-cell contact. The dynamics of plant growth, morphogenesis and differentiation require concomitant modifications of the walls. A class of enzymes known as xyloglucan endotransglycosylases have the potential to enzymatically modify wall components, but although their biochemical activity has been defined, the physiological roles of xyloglucan endotransglycosylases remain undefined. Xyloglucan endotransglycosylases are encoded by large gene families, and in an attempt to clarify their physiological role, the diverse regulation of the genes and properties of the proteins are being determined. PMID- 10462770 TI - The 14-3-3 proteins: cellular regulators of plant metabolism. AB - Signal transduction and enzyme regulation are known to occur via phosphorylation, but it is becoming increasingly apparent that phosphorylation might be only a necessary preamble to regulation. In many cases, the phosphorylated target protein must associate with a specialized adapter protein, known as 14-3-3, to complete the regulatory action. There are several prominent examples of 14-3-3 participation in plant regulatory events, including the regulation of plasma membrane H+-ATPase, nitrate reductase and sucrose phosphate synthase. However, emerging data on 14-3-3s in the nucleus might extend the roles for 14-3-3s well beyond the regulation of cytoplasmic enzymes. PMID- 10462771 TI - Water ascent in plants: do ongoing controversies have a sound basis? AB - The cohesion-tension theory of the ascent of sap in plants is fundamental to the understanding of water movement in plants. According to the theory, water is pulled upwards by high tensions (low negative pressures) created in the xylem vessels and tracheids of higher plants by the evaporation of water vapour from leaves. However, much lower tensions (less negative pressures) have been found from direct measurements using a pressure probe. These do not appear to be compatible with the cohesion-tension theory. As a consequence, the validity of the cohesion-tension theory has been questioned and alternative mechanisms for sap ascent have been proposed. Recent experiments show that the conclusions drawn from the pressure probe work were premature. New direct measurements of xylem pressure support the cohesion-tension theory and the previous indirect measurements of xylem pressure. PMID- 10462772 TI - Analysis of the N-glycosylation of recombinant glycoproteins produced in transgenic plants. PMID- 10462774 TI - Targeted transgenics from the creators of Dolly. PMID- 10462773 TI - Contaminated ultrasound probes: a possible source of nosocomial infections. PMID- 10462776 TI - 1998 William P. Yant Award Lecture. What is the correlation between scientific and social challenges in occupational hygiene? PMID- 10462777 TI - Systemic uptake and clearance of chloroform by hairless rats following dermal exposure: II. Absorption of the neat solvent. AB - Blood concentrations of chloroform were monitored after exposing small areas (approximately 5.5 cm2) of the backs of hairless rats to liberal excesses of the solvent for either 1, 3, or 8 min. The amounts absorbed were quantified by comparing areas-under-the-curves (AUCs) of blood concentration versus time plots to the AUC obtained on infusing an aqueous chloroform solution of known concentration for 30 min (positive control). Chloroform penetrated the dermal barrier rapidly, the skin's horny layer and the deeper skin tissues acting as reservoirs for chloroform only for short durations. Evaporative and physiological clearance from these reservoirs was rapid once the chloroform was removed from the surface. Pressure of the template used to confine the exposure affected uptake. For blood levels, the time to reach the maximum blood concentration increased with increased exposure duration. Amounts absorbed also depended on exposure duration. Blood level profiles indicated systemic uptake of chloroform following a 3-min exposure was about 1.3-fold higher than for a 1-min exposure (not significant), while the 8-min exposure produced an AUC roughly 3.8-fold higher than found at 3 min (p = 0.026). Chloroform is rapidly cleared from rat blood (terminal elimination rate constant = 0.009/min). Calculations indicated that its absorption from these area-limited exposures far exceeds that which would be absorbed had the chloroform been presented to the skin as a saturated aqueous solution. PMID- 10462778 TI - Lead-based paint testing technologies: summary of an EPA/HUD field study. AB - The U.S. Environmental Protection Agency and the U.S. Department of Housing and Urban Development sponsored a field study of portable technologies for testing for lead in paint in three U.S. cities in 1993. Six chemical test kits and six X ray fluorescence instruments, which represented the two main types of portable technologies available for residential lead testing at that time, were evaluated. Painted building components in single-family and multifamily housing units were selected to assess the performance of these products under real-world conditions. The study found that the chemical test kits were not effective in distinguishing lead-based paint, as defined by federal standards, from nonlead based paint. The X-ray fluorescence instruments were, under certain circumstances, found to be effective. The study filled an informational gap about the accuracy and precision of the portable lead-testing technologies. This article describes the design of the study and its major findings. PMID- 10462779 TI - Determination of glycols in air: development of sampling and analytical methodology and application to theatrical smokes. AB - Glycol-based fluids are used in the production of theatrical smokes in theaters, concerts, and other stage productions. The fluids are heated and dispersed in aerosol form to create the effect of a smoke, mist, or fog. There have been reports of adverse health effects such as respiratory irritation, chest tightness, shortness of breath, asthma, and skin rashes. Previous attempts to collect and quantify the aerosolized glycols used in fogging agents have been plagued by inconsistent results, both in the efficiency of collection and in the chromatographic analysis of the glycol components. The development of improved sampling and analytical methodology for aerosolized glycols was required to assess workplace exposures more effectively. An Occupational Safety and Health Administration versatile sampler tube was selected for the collection of ethylene glycol, propylene glycol, 1,3-butylene glycol, diethylene glycol, triethylene glycol, and tetraethylene glycol aerosols. Analytical methodology for the separation, identification, and quantitation of the six glycols using gas chromatography/flame ionization detection is described. Limits of detection of the glycol analytes ranged from 7 to 16 micrograms/sample. Desorption efficiencies for all glycol compounds were determined over the range of study and averaged greater than 90%. Storage stability results were acceptable after 28 days for all analytes except ethylene glycol, which was stable at ambient temperature for 14 days. Based on the results of this study, the new glycol method was published in the NIOSH Manual of Analytical Methods. PMID- 10462780 TI - Effect of respirator inspiratory resistance level on constant load treadmill work performance. AB - Respirator inspiratory resistance can affect performance times, especially when the experiment is optimized to elicit respiratory stress. Twelve subjects performed on a treadmill at constant speeds and grades chosen to result in performance times of 5-15 min. Six levels of inspiratory resistance were used, ranging from 0.78 to 7.64 cm H2O.sec/L. The results showed that performance times decrease linearly with resistance level, and no threshold resistance value is apparent. Inspiratory resistance also induces hypoventilation, with lower minute volumes and lower oxygen consumption values at higher resistances. These trends are also linear. From these results, there is no value for inspiratory resistance that can be given as a design goal. Other parameters such as weight and space may dictate filter resistance values, and these, in turn, will lead to determined performance degradations. PMID- 10462781 TI - A pilot study for monitoring changes in the microbiological component of metalworking fluids as a function of time and use in the system. AB - This article describes the results of a pilot study to examine changes in the biological component of metalworking fluids (MWF) as a function of use. Fluid samples were taken from two newly charged systems, designated BT-7415 and BT 7707, at 1-week intervals for 8 weeks and characterized with respect to the kinds and numbers of bacteria present and presence of soluble protein in cell-free supernatants. In addition, lipid extracts of pelleted cells from fluids in BT 7415 were examined by gas chromatography/mass spectroscopy for the kinds and relative amounts of phospholipid fatty acids (PLFA) present. A total of 19 different bacterial species was cultured and identified, more than half (12/19) of which were gram-negative. Total colony-forming units (CFU) reached levels of 2.2 x 10(3)/mL in BT-7415 and 2.4 x 10(5)/mL in BT-7707. The most common genus isolated was Pseudomonas. Estimations of cell numbers based on total biomass from PLFA in samples from BT-7415 indicated 1.1 x 10(7)/mL after 8 weeks of use. Both the numbers of PLFA identified and the amounts of each detected in BT-7415 increased as the fluids were used. The chromatograms were dominated by two fatty acids, the amounts of which increased with time. These fatty acids, 18:2 omega 6 and 18:1 omega 9c, are not commonly associated with pseudomonads. This suggests that there is an important component of the biological consortium in MWF is not being detected by currently used culture techniques. There was no soluble protein detected in any of the samples from either system. PMID- 10462782 TI - Evaluation of exposures to fluorocarbon 113 in a horizontal and a vertical laminar airflow clean room. AB - Exposures to 1,1,2-trichloro-1,2,2-trifluoroethane or fluorocarbon (FC) 113 were evaluated in a horizontal laminar airflow (HLAF) clean room and a vertical laminar airflow (VLAF) clean room. A full period consecutive samples measurement strategy was employed. Data were used to calculate 8-hour time-weighted averages (8-TWA) for major work groups and to characterize exposures associated with specific cleaning tasks. The MIRAN 1B infrared analyzer was used to estimate peak concentrations. In the HLAF clean room, 8-TWAs ranged from 193 to 439 ppm; in the VLAF clean room, 8-TWAs ranged from 110 to 935 ppm. These levels were below the current Occupational Safety and Health Administration permissible exposure limit and the National Institute for Occupational Safety and Health (NIOSH) recommended exposure limit for FC 113 of 1000 ppm. Short-term sample concentrations ranged from 104 ppm (inspection) to 1080 ppm (assembly) in the HLAF clean room and 51 ppm (packaging)-3380 ppm (flushing) in the VLAF clean room. In the VLAF clean room, several short-term concentrations measured during the flushing task--1421 ppm and 2522 ppm--were above the NIOSH short-term exposure limit (STEL) of 1250 ppm. These data suggest the possibility that the STEL may be exceeded for tasks involving direct work with liquid FC 113. Peak exposure levels may be reduced by modification of worker position in the HLAF clean room and by use of open wire tables in the VLAF clean room. PMID- 10462783 TI - A comparison of physiological responses to two types of particle barrier, vapor permeable clothing ensembles. AB - Chemical protective clothing (PC) use while working results in elevated rectal temperatures (Tre) that limit work time. Particle barrier, vapor permeable (PBVP) PCs allow workers to cool themselves by evaporating some sweat. The purpose of this study was to compare the effects on worker productivity of two types of PBVP suits, a Kleenguard (PPPC) (Kimberly Clark), and a Tyvek (PEPC) (DuPont) suit. Fifteen males in a repeated measures design performed four work tests consisting of a walk/arm curl combination at a time-weighted work rate of 1.0 L/min (300 kcal/hr), two in a wet bulb globe temperature (WBGT) of 26 degrees C and two in a WBGT of 18 degrees C, with subjects wearing each suit once in each environment. No significant difference (p > 0.05) was observed between the suits at 18 degrees C WBGT, but a significant difference was found (p < 0.05) between the suits, with the PPPC having a lower Tre in the WBGT = 26 degrees C at the 80th, 100th, and 120th min. A significant difference (p < .05) was also seen in the 26 degrees C WBGT with the PPPC resulting in a lower heart rate (HR) at the 40th, 60th, 80th, 100th, and 120th min and rate of perceived exertion (RPE) at the 75th, 90th, and 120th min. Additionally, a significant difference (p < .05) was seen between PEPC and PPPC for Tre, delta Tre, mean skin temp (mTsk), delta mTsk, and HR, each regressed against time in the 26 degrees C WBGT. Twelve of the 15 subjects also reported feeling cooler in the PPPC versus the PEPC in either WBGT environment. PMID- 10462784 TI - Evaluation and field calibration of the Miniram PDM-3 aerosol monitor for measuring respirable and total coal dust. AB - The MIE Miniram PDM-3 is a real-time aerosol dust monitor designed to measure dust based on Mie scattering. It has an optional in-line filter that, when attached to a constant flow air sampling pump, allows a gravimetric air sample to be collected from the same air stream that passes through the Miniram sensing chamber. This study compared real-time Miniram digital respirable and total dust readings with concentrations from an in-line 5 microns polyvinyl chloride filter connected to a constant flow pump and with results from traditional respirable and total dust samples. Area samples were collected at three coal-fired power generating facilities over a 2-month period. Traditional respirable dust concentrations ranged from 0.04 to 10.8 mg/m3. In the monitored range of concentrations the Miniram respirable concentrations and the in-line respirable dust concentrations were not significantly different (p > or = 0.05), nor were the Miniram in-line filter concentrations and the side-by-side respirable dust concentrations significantly different (p > or = 0.05). However, the Miniram respirable concentrations and the traditional respirable concentrations were significantly different (p = 0.02). The Statistical Analysis System (SAS) procedure CALIS, which meets the statistical requirements for developing calibration lines for two variables each measured with error, is used to develop field calibration curves for the comparisons between direct-reading instrument concentrations and concentrations from traditional National Institute for Occupational Safety and Health methods for total and respirable dust. PMID- 10462785 TI - Prediction of respiratory distress during maximal physical exercise: the role of trait anxiety. AB - Industrial respirators offer protection for men and women who are required to work in toxic and oxygen-deficient environments. Major advances continue to be made with respect to improved face mask designs and modes of protection, yet only recently have efforts been directed toward the development of criteria for use in evaluating individuals' psychological fitness to wear respirators. The purpose of the present investigation was to confirm (i.e., replicate) an earlier finding, using a simplified protocol, that an individual's tendency to experience respiratory distress during exercise can be predicted. In the present experiment, an independent sample of 38 subjects underwent a maximal exercise test. It was predicted that subjects with elevated trait anxiety scores would experience respiratory distress when required to perform heavy physical exercise using a full-facepiece, air-line supplied, pressure-demand respirator. The prediction of respiratory distress was accurate in 34 of 38 cases (89.5%). It was concluded that an objective measure of trait anxiety can be used to identify those individuals who are most likely to experience distress while performing maximal physical exercise and using a pressure-demand respirator. PMID- 10462786 TI - Tank truck driver exposure to vapors from oxygenated or reformulated gasolines during loading and unloading. AB - Tank truck drivers' exposure to gasoline vapors was studied by collecting breathing zone samples during loading and unloading of gasoline. The field studies were conducted at three dispatches and at seven service stations in Finland. The gasolines included in the study (95, 98, 99 research octane number, RON) were of reformulated or oxygenated grade containing about 2% (w/w) oxygen and 0.5-1.5% (v/v) benzene. The sampling times ranged from 16 to 57 min (mean 35 min), and time-weighted average concentrations for a 30-min period were calculated. Using the time-adjusted values, geometric mean concentrations (GM) were calculated for three periods of dispatch measurements (n = 15,20,7) and a period of unloading measurements at service stations (n = 7). The GM for methyl tert-butyl ether ranged from 0.95 to 7.3 mg/m3 and that for tert-amyl methyl ether from 0.30 to 1.1 mg/m3. The GM concentrations of hexane, benzene, and toluene were in the range of 0.25-2.3 mg/m3, 0.15-0.28 mg/m3, and 0.73-1.7 mg/m3, respectively. Multiple regression analysis yielded an r2 value of 0.98 for the daily mean concentration of toluene and correspondingly 0.94 for benzene when daily wind speed (0.1-3.7 m/sec) and daily air temperature (-7.4(-)+17.2 degrees C) were used as independent variables. The average number of gasoline loads per tank truck was 2.5, corresponding to 23,000 L of gasoline. PMID- 10462787 TI - An axisymmetric poroelastic finite element formulation. AB - In the past, various two- and three-dimensional Cartesian, poroelastic finite element formulations have been proposed and demonstrated. Here an axisymmetric formulation of a poroelastic finite element is presented. The intention of this work was to develop a finite element formulation that could easily and efficiently model axisymmetric sound propagation in circular structures having arbitrary, axially dependent radii, and that are lined or filled with elastic porous sound absorbing materials such as foams. The formulation starts from the Biot equations for an elastic porous material expressed explicitly in axisymmetric form. By following a standard finite element development, a u-U formulation results. Procedures for coupling the axisymmetric elements to an adjacent acoustical domain are described, as are the boundary conditions appropriate for unfaced foams. Calculations described here show that the present formulation yields predictions as accurate as a Cartesian, three-dimensional model in much reduced time. Predictions made using the present model are also compared with measurements of sound transmission through cylindrical foam plugs, and the predicted results are shown to agree well with the measurements. Good agreement was also found in the case of sound transmission through a conical foam plug. PMID- 10462788 TI - Broadband performance of an active headrest. AB - This paper presents a study of the attenuation of broadband random acoustic disturbances, when using a feedback active headrest system, as originally suggested by Olson and May. Previous studies showed that a practical active headrest can be designed for tonal disturbances using feedforward controllers. However, many applications, such as jet aircraft and cars, require feedback systems to control random disturbances over a wide frequency bandwidth. In this work, robust feedback controllers are designed to control broadband random disturbances in the low-frequency range based on measured data from a laboratory headrest system. The results show that a practically useful performance can be achieved, but only if the controller is designed to minimize the pressure at a "virtual microphone" close to the listener's ears, and that the performance is maintained reasonably well with movements of the listener's head. The paper emphasizes the importance of both the acoustics and the control in the design of broadband active headrest systems. PMID- 10462789 TI - Locating animals from their sounds and tomography of the atmosphere: experimental demonstration. AB - Calling animals are located using widely distributed receivers, and the sounds from the animals are used to map the sound speed and wind fields by means of tomography. In particular, two Red-Winged Blackbirds Agelaius phoeniceus are correctly located within a meter using recordings from five receivers spread over a 20 by 30 m region. The demonstration hinges on two new developments. First, a new algorithm for blindly estimating the impulse response of the channel is shown capable of estimating the differences in the time of first arrivals at two receivers. Since it is known that the first arrivals travel along nearly straight paths, the difference in time constrains the animal's location to a hyperboloid, and the animal is located by intersecting hyperboloids from many pairs of receivers. Second, in order to accurately find the intersection point and map the sound speed and wind fields using tomography, a nonlinear equation is solved. The new algorithm for blindly estimating the impulse response of a channel offers a new way for locating sounds and making tomographic maps of the environment without any requirement for a model for the propagation of sound such as is needed for focalization and matched field processing. PMID- 10462790 TI - Longitudinal endolymph movements and endocochlear potential changes induced by stimulation at infrasonic frequencies. AB - The inner ear is continually exposed to pressure fluctuations in the infrasonic frequency range (< 20 Hz) from external and internal body sources. The cochlea is generally regarded to be insensitive to such stimulation. The effects of stimulation at infrasonic frequencies (0.1 to 10 Hz) on endocochlear potential (EP) and endolymph movements in the guinea pig cochlea were studied. Stimuli were applied directly to the perilymph of scala tympani or scala vestibuli of the cochlea via a fluid-filled pipette. Stimuli, especially those near 1 Hz, elicited large EP changes which under some conditions exceeded 20 mV in amplitude and were equivalent to a cochlear microphonic (CM) response. Accompanying the electrical responses was a cyclical, longitudinal displacement of the endolymph. The amplitude and phase of the CM varied according to which perilymphatic scala the stimuli were applied to and whether a perforation was made in the opposing perilymphatic scala. Spontaneously occurring middle ear muscle contractions were also found to induce EP deflections and longitudinal endolymph movements comparable to those generated by perilymphatic injections. These findings suggest that cochlear fluid movements induced by pressure fluctuations at infrasonic frequencies could play a role in fluid homeostasis in the normal state and in fluid disturbances in pathological states. PMID- 10462791 TI - Medial efferent effects on auditory-nerve responses to tail-frequency tones. I. Rate reduction. AB - One way medial efferents are thought to inhibit responses of auditory-nerve fibers (ANFs) is by reducing the gain of the cochlear amplifier thereby reducing motion of the basilar membrane. If this is the only mechanism of medial efferent inhibition, then medial efferents would not be expected to inhibit responses where the cochlear amplifier has little effect, i.e., at sound frequencies in the tails of tuning curves. Inhibition at tail frequencies was tested for by obtaining randomized rate-level functions from cat ANFs with high characteristic frequencies (CF > or = 5 kHz), stimulated with tones two or more octaves below CF. It was found that electrical stimulation of medial efferents can indeed inhibit ANF responses to tail-frequency tones. The amplitude of efferent inhibition depended on both sound level (largest near to threshold) and frequency (largest two to three octaves below CF). On average, inhibition of high-CF ANFs responding to 1 kHz tones was around 5 dB. Although an efferent reduction of basilar-membrane motion cannot be ruled out as the mechanism producing the inhibition of ANF responses to tail frequency tones, it seems more likely that efferents produce this effect by changing the micromechanics of the cochlear partition. PMID- 10462792 TI - The normalized interaural correlation: accounting for NoS pi thresholds obtained with Gaussian and "low-noise" masking noise. AB - Recently, Eddins and Barber [J. Acoust. Soc. Am. 103, 2578-2589 (1998)] and Hall et al. [J. Acoust. Soc. Am. 103, 2573-2577 (1998)] independently reported that greater masking of interaurally phase-reversed (S pi) tones was produced by diotic low-noise noise than by diotic Gaussian noise. Based on quantitative analyses, Eddins and Barber suggested that their results could not be accounted for by assuming that listeners' judgments were based on constant-criterion changes in the normalized interaural correlation produced by adding the S pi signal to the diotic masker. In particular, they showed that a model like the one previously employed by Bernstein and Trahiotis [J. Acoust. Soc. Am. 100, 3774 3784 (1996)] predicted an ordering of thresholds between the conditions of interest that was opposite to that observed. Bernstein and Trahiotis computed the normalized interaural correlation subsequent to half-wave, square-law rectification and low-pass filtering, the parameters of which were chosen to mimic peripheral auditory processing. In this report, it is demonstrated that augmenting the model by adding a physiologically valid stage of "envelope compression" prior to rectification and low-pass filtering provides a remedy. The new model not only accounts for the data obtained by Eddins and Barber (and the similar data obtained by Hall et al.), but also does not diminish the highly successful account of the comprehensive set of data that gave rise to the original form of the model. Therefore, models based on the computation of the normalized interaural correlation appear to remain valid because they can account, both quantitatively and qualitatively, for a wide variety of binaural detection and discrimination data. PMID- 10462793 TI - Proportional frequency compression of speech for listeners with sensorineural hearing loss. AB - This study examined proportional frequency compression as a strategy for improving speech recognition in listeners with high-frequency sensorineural hearing loss. This method of frequency compression preserved the ratios between the frequencies of the components of natural speech, as well as the temporal envelope of the unprocessed speech stimuli. Nonsense syllables spoken by a female and a male talker were used as the speech materials. Both frequency-compressed speech and the control condition of unprocessed speech were presented with high pass amplification. For the materials spoken by the female talker, significant increases in speech recognition were observed in slightly less than one-half of the listeners with hearing impairment. For the male-talker materials, one-fifth of the hearing-impaired listeners showed significant recognition improvements. The increases in speech recognition due solely to frequency compression were generally smaller than those solely due to high-pass amplification. The results indicate that while high-pass amplification is still the most effective approach for improving speech recognition of listeners with high-frequency hearing loss, proportional frequency compression can offer significant improvements in addition to those provided by amplification for some patients. PMID- 10462794 TI - Pitch shifts of mistuned partials: a time-domain model. AB - Mistuning one partial of a complex harmonic tone makes that partial easier to hear as a tone separate from the complex. At the same time, two pitch shifts may be observed. First, the low pitch of the complex is shifted in the direction of the mistuning, as if it were "pulled" by the partial. Second, the mistuning of the partial is perceptually exaggerated, as if the pitch of the partial were "pushed" away from the harmonic series defined by the complex. This paper shows how the latter effect can emerge within a hypothetical neural circuit. The circuit involves a gating neuron fed by three pathways, one direct and excitatory and the other two delayed and inhibitory. The neuron responds to any excitatory input spike unless it is accompanied by an inhibitory input spike on either delayed input, thus acting as a kind of "anticoincidence counter." The first delay is fixed and tuned to the period of the background harmonic complex. Its purpose is to weaken correlates of in-tune components and allow the mistuned partial to stand out. The second delay is variable and used to estimate the period of the mistuned partial, by searching for a minimum output as a function of delay. With an appropriate choice of parameters, the estimate is subject to shifts that are of the same sign as the mistuning and that peak at about 4% mistuning and decrease beyond, as observed experimentally. PMID- 10462795 TI - Further evaluation of a model of loudness perception applied to cochlear hearing loss. AB - This paper describes further tests of a model for loudness perception in people with cochlear hearing loss. It is assumed that the hearing loss (the elevation in absolute threshold) at each audiometric frequency can be partitioned into a loss due to damage to outer hair cells (OHCs) and a loss due to damage to inner hair cells (IHCs) and/or neurons. The former affects primarily the active mechanism that amplifies the basilar membrane (BM) response to weak sounds. It is modeled by increasing the excitation level required for threshold, which results in a steeper growth of specific loudness with increasing excitation level. Loss of frequency selectivity, which results in broader excitation patterns, is also assumed to be directly related to the OHC loss. IHC damage is modeled by an attenuation of the calculated excitation level at each frequency. The model also allows for the possibility of complete loss of IHCs or functional neurons at certain places within the cochlea ("dead" regions). The parameters of the model (OHC loss at each audiometric frequency, plus frequency limits of the dead regions) were determined for three subjects with unilateral cochlear hearing loss, using data on loudness matches between sinusoids presented alternately to their two ears. Further experiments used bands of noise that were either 1 equivalent rectangular bandwidth (ERB) wide or 6-ERBs wide, centered at 1 kHz. Subjects made loudness matches for these bands of noise both within ears and across ears. The model was reasonably accurate in predicting the results of these matches without any further adjustment of the parameters. PMID- 10462796 TI - Modulation masking produced by beating modulators. AB - This study examined whether "modulation masking" could be produced by temporal similarity of the probe and masker envelopes, even when the masker envelope did not contain a spectral component close to the probe frequency. Both masker and probe amplitude modulation were applied to a single 4-kHz sinusoidal or narrow band noise carrier with a level of 70 dB SPL. The threshold for detecting 5-Hz probe modulation was affected by the presence of a pair of masker modulators beating at a 5-Hz rate (40 and 45 Hz, 50 and 55 Hz, or 60 and 65 Hz). The threshold was dependent on the phase of the probe modulation relative to the beat cycle of the masker modulators; the threshold elevation was greatest (12-15 dB for the sinusoidal carrier and 9-11 dB for the noise carrier, expressed as 20 log m) when the peak amplitude of the probe modulation coincided with a peak in the beat cycle. The maximum threshold elevation of the 5-Hz probe produced by the beating masker modulators was 7-12 dB greater than that produced by the individual components of the masker modulators. The threshold elevation produced by the beating masker modulators was 2-10 dB greater for 5-Hz probe modulation than for 3- or 7-Hz probe modulation. These results cannot be explained in terms of the spectra of the envelopes of the stimuli, as the beating masker modulators did not produce a 5-Hz component in the spectra of the envelopes. The threshold for detecting 5-Hz probe modulation in the presence of 5-Hz masker modulation varied with the relative phase of the probe and masker modulation. The pattern of results was similar to that found with the beating two-component modulators, except that thresholds were highest when the masker and probe were 180 degrees out of phase. The results are consistent with the idea that nonlinearities within the auditory system introduce distortion in the internal representation of the envelopes of the stimuli. In the case of two-component beating modulators, a weak component is introduced at the beat rate, and it has an amplitude minimum when the beat cycle is at its maximum. The results could be fitted well using two models, one based on the concept of a sliding temporal integrator and one based on the concept of a modulation filter bank. PMID- 10462797 TI - Correlational analysis of acoustic cues for the discrimination of auditory motion. AB - The sound of a source moving in a straight path and passing directly in front of the listener on the azimuthal plane was synthesized over headphones to include three dynamic cues for motion: Doppler effect, overall intensity, and interaural time difference. Discriminability of a change in displacement, velocity, and acceleration of this source was measured using a standard two-interval, forced choice procedure. In each case, the relative reliance or weight given to the three acoustic cues was estimated from correlations of the listener's response with small independent pertubations imposed on cues from trial to trial. Group estimates of threshold agreed well with results from past studies, while the obtained pattern of weights depended on the individual, starting velocity, and discrimination task. For the discrimination of displacement at moderate velocity (10 m/s), responses were most highly correlated with intensity or interaural time difference. For the discrimination of velocity and, to a lesser extent, acceleration, responses were most highly correlated with Doppler effect. At higher velocity (50 m/s) responses in all discrimination tasks were most strongly correlated with Doppler effect with few exceptions. Randomizing source spectrum or roving distance of the source from trial to trial did not significantly affect the pattern of results. The results suggest that motion perception is mutable, and not in all cases based on a single invariant acoustic cue. PMID- 10462798 TI - Can macaques perceive place of articulation from formant transition information? AB - An important problem in speech perception is to determine how humans extract the perceptually invariant place of articulation information in the speech wave across variable acoustic contexts. Although analyses have been developed that attempted to classify the voiced stops /b/ versus /d/ from stimulus onset information, most of the human perceptual research to date suggests that formant transition information is more important than onset information. The purpose of the present study was to determine if animal subjects, specifically Japanese macaque monkeys, are capable of categorizing /b/ versus /d/ in synthesized consonant-vowel (CV) syllables using only formant transition information. Three monkeys were trained to differentiate CV syllables with a "go-left" versus a "go right" label. All monkeys first learned to differentiate a /za/ versus /da/ manner contrast and easily transferred to three new vowel contexts /[symbol: see text], epsilon, I/. Next, two of the three monkeys learned to differentiate a /ba/ versus /da/ stop place contrast, but were unable to transfer it to the different vowel contexts. These results suggest that animals may not use the same mechanisms as humans do for classifying place contrasts, and call for further investigation of animal perception of formant transition information versus stimulus onset information in place contrasts. PMID- 10462799 TI - The role of spectral and periodicity cues in auditory stream segregation, measured using a temporal discrimination task. AB - In a previous paper, it was shown that sequential stream segregation could be based on both spectral information and periodicity information, if listeners were encouraged to hear segregation [Vliegen and Oxenham, J. Acoust. Soc. Am. 105, 339 346 (1999)]. The present paper investigates whether segregation based on periodicity information alone also occurs when the task requires integration. This addresses the question: Is segregation based on periodicity automatic and obligatory? A temporal discrimination task was used, as there is evidence that it is difficult to compare the timing of auditory events that are perceived as being in different perceptual streams. An ABA ABA ABA... sequence was used, in which tone B could be either exactly at the temporal midpoint between two successive tones A or slightly delayed. The tones A and B were of three types: (1) both pure tones; (2) both complex tones filtered through a fixed passband so as to contain only harmonics higher than the 10th, thereby eliminating detectable spectral differences, where only the fundamental frequency (f0) was varied between tones A and B; and (3) both complex tones with the same f0, but where the center frequency of the spectral passband varied between tones. Tone A had a fixed frequency of 300 Hz (when A and B were pure tones) or a fundamental frequency (f0) of 100 Hz (when A and B were complex tones). Five different intervals, ranging from 1 to 18 semitones, were used. The results for all three conditions showed that shift thresholds increased with increasing interval between tones A and B, but the effect was largest for the conditions where A and B differed in spectrum (i.e., the pure-tone and the variable-center-frequency conditions). The results suggest that spectral information is dominant in inducing (involuntary) segregation, but periodicity information can also play a role. PMID- 10462800 TI - Modulation gap detection: effects of modulation rate, carrier separation, and mode of presentation. AB - Modulation gap detection (MGD) is a procedure that measures the sensitivity to an interruption in the modulation pattern imposed upon one or more carrier frequencies. The MGD task was developed to test conditions where a temporal event traverses frequency, but without a concomitant interruption in the spectral continuity of the stimulus. This contrasts with across-frequency gap detection where there is an inherent spectral discontinuity associated with the temporal gap, and where there is a marked decline in performance when the markers of the temporal gap are widely separated in frequency. The purpose of this study was to test the hypothesis that a wideband temporal analysis will be facilitated if there exists a spectral continuity throughout the temporal event. Experiment 1 established the procedure of MGD and indicated that a modulation rate of 8 Hz was optimal for the task. Experiment 2 showed that performance declined markedly when the carrier frequencies of the modulation markers were widely separated in frequency. This finding indicates that spectral continuity across the temporal event is not a sufficient prerequisite for the auditory system to undertake a wideband temporal analysis. Experiment 3 revealed that dichotic MGD also results in poor performance, similar to that seen for widely separated carrier frequencies in the monaural case. This supports the hypothesis that the "channels" across which temporal events are poorly processed do not necessarily correspond to peripheral frequency channels. PMID- 10462801 TI - Auditory temporal integration in the rhesus macaque (Macaca mulatta). AB - Temporal integration for pure tones was examined in two rhesus macaques. The subjects were required to respond to a brief sound (a tone burst) that deviated from a previous series of sounds (noise bursts) on a trial (a deviant-stimulus detection paradigm). Psychometric functions and thresholds were determined from correct detections (hit proportions) alone, and from d' scores. Two models describing the decline in threshold as a function of stimulus duration, one a power function the other an exponential, were tested against the data. When the decline (slope) in threshold per log stimulus duration is used as a rate measure, our results yield a lower estimate of temporal integration rate in rhesus than did a previous study [Clack, J. Acoust. Soc. Am. 40, 1140-1146 (1966)]. Both studies, however, gave slope estimates of integration rate that were higher than in most other species. Comparison of the models using data from several species, revealed that the exponential, but not the power model, could account for two sources of variation in threshold measurement. One source is due to the range across threshold as a function of duration (the linear rate component), and is described by the constant of proportionality Ik in the model. The other source of variation arises from the rate of decline within this range (the nonlinear rate component), and is described by the time constant tau. In terms of this model, differences in rate estimates between Clack's study and ours (and between rhesus and other species) are primarily due to the linear component. The nonlinear rate component was about equal for our study and Clack's (tau = approximately 150 ms): a time constant that is just slightly larger (indicating a rate of temporal integration slightly slower) than for most other species examined. PMID- 10462802 TI - Psychometric functions for gap detection in noise measured from young and aged subjects. AB - Psychometric functions for gap detection of temporal gaps in wideband noise were measured in a "yes/no" paradigm from normal-hearing young and aged subjects with closely matched audiograms. The effects of noise-burst duration, gap location, and uncertainty of gap location were tested. A typical psychometric function obtained in this study featured a steep slope, which was independent of most experimental conditions as well as age. However, gap thresholds were generally improved with increasing duration of the noise burst for both young and aged subjects. Gap location and uncertainty had no significant effects on the thresholds for the young subjects. For the aged subjects, whenever the gap was sufficiently away from the onset or offset of the noise burst, detectability was robust despite uncertainty about the gap location. Significant differences between young and aged subjects could be observed only when the gap was very close to the signal onset and offset. PMID- 10462803 TI - The contribution of static and dynamically varying ITDs and IIDs to binaural detection. AB - This paper investigates the relative contribution of various interaural cues to binaural unmasking in conditions with an interaurally in-phase masker and an out of-phase signal (MoS pi). By using a modified version of multiplied noise as the masker and a sinusoid as the signal, conditions with only interaural intensity differences (IIDs), only interaural time differences (ITDs), or combinations of the two were realized. In addition, the experimental procedure allowed the presentation of specific combinations of static and dynamically varying interaural differences. In these conditions with multiplied noise as masker, the interaural differences have a bimodal distribution with a minimum at zero IID or ITD. Additionally, by using the sinusoid as masker and the multiplied noise as signal, a unimodal distribution of the interaural differences was realized. Through this variation in the shape of the distributions, the close correspondence between the change in the interaural cross correlation and the size of the interaural differences is no longer found, in contrast to the situation for a Gaussian-noise masker [Domnitz and Colburn, J. Acoust. Soc. Am. 59, 598-601 (1976)]. When analyzing the mean thresholds across subjects, the experimental results could not be predicted from parameters of the distributions of the interaural differences (the mean, the standard deviation, or the root-mean square value). A better description of the subjects' performance was given by the change in the interaural correlation, but this measure failed in conditions which produced a static interaural intensity difference. The data could best be described by using the energy of the difference signal as the decision variable, an approach similar to that of the equalization and cancellation model. PMID- 10462804 TI - A comparison of monotic and dichotic complex-tone pitch perception in listeners with hearing loss. AB - The perception of fundamental pitch for two-harmonic complex tones was examined in musically experienced listeners with cochlear-based high-frequency hearing loss. Performance in a musical interval identification task was measured as a function of the average rank of the lowest harmonic for both monotic and dichotic presentation of the harmonics at 14 dB Sensation Level. Listeners with hearing loss demonstrated excellent musical interval identification at low fundamental frequencies and low harmonic numbers, but abnormally poor identification at higher fundamental frequencies and higher average ranks. The upper frequency limit of performance in the listeners with hearing loss was similar in both monotic and dichotic conditions. These results suggest that something other than frequency resolution per se limits complex-tone pitch perception in listeners with hearing loss. PMID- 10462805 TI - The perceptual effects of current pulse duration in electrical stimulation of the auditory nerve. AB - In cochlear implants employing pulsatile stimulation, loudness is controlled by current amplitude and/or pulse duration. Five experiments were conducted with cochlear implantees to investigate the hypothesis that perceptual effects other than loudness result from changes in pulse duration for durations from 50 to 266 microseconds. In experiment 1, five subjects' ability to discriminate equally loud pulse trains employing differing pulse durations was measured at four electrode positions. In 11 of the 20 cases, subjects could significantly discriminate these stimuli. In experiments 2 and 3, discrimination was measured of dual-electrode stimuli which differed in overall temporal pattern but had an equal temporal pattern on each of the individual electrodes (separated by 0 to 9 mm). Discrimination was compared for stimuli employing short or long pulse durations and, in experiment 3, employing different pulse durations on each electrode. When the pulse duration was longer, six out of seven subjects could either combine temporal information across electrode positions at wider electrode separations (experiment 2) or had better discrimination at the same electrode separation (experiment 3). This result was consistent with the hypothesis that longer pulse durations result in a greater spread of excitation than equally loud stimuli using shorter pulse durations. In experiment 4, pulse rate discrimination was compared for stimuli with differing pulse durations, and in four out of five subjects, there was no effect of pulse duration. Finally, the dB change in current per doubling of pulse duration for threshold and equally loud stimuli was calculated for nine subjects (52 electrodes). Values ranged from -5.9 to -2.0 dB/doubling, and were significantly correlated with the absolute intensity of the stimulus. This result was hypothesized to be due to a relationship between the neural charging characteristics and the distance of the excited neural elements from the electrode. PMID- 10462806 TI - Constrained adaptation for feedback cancellation in hearing aids. AB - In feedback cancellation in hearing aids, an adaptive filter is used to model the feedback path. The output of the adaptive filter is subtracted from the microphone signal to cancel the acoustic and mechanical feedback picked up by the microphone, thus allowing more gain in the hearing aid. In general, the feedback cancellation filter adapts on the hearing-aid input signal, and signal cancellation and coloration artifacts can occur for a narrow-band input. In this paper, two procedures for LMS adaptation with a constraint on the magnitude of the adaptive weight vector are derived. The constraints greatly reduce the probability that the adaptive filter will cancel a narrow-band input. Simulation results are used to demonstrate the efficacy of the constrained adaptation. PMID- 10462807 TI - On the lingual organization of the German vowel system. AB - A hybrid PARAFAC and principal-component model of tongue configuration in vowel production is presented, using a corpus of German vowels in multiple consonant contexts (fleshpoint data for seven speakers at two speech rates from electromagnetic articulography). The PARAFAC approach is attractive for explicitly separating speaker-independent and speaker-dependent effects within a parsimonious linear model. However, it proved impossible to derive a PARAFAC solution of the complete dataset (estimated to require three factors) due to complexities introduced by the consonant contexts. Accordingly, the final model was derived in two stages. First, a two-factor PARAFAC model was extracted. This succeeded; the result was treated as the basic vowel model. Second, the PARAFAC model error was subjected to a separate principal-component analysis for each subject. This revealed a further articulatory component mainly involving tongue blade activity associated with the flanking consonants. However, the subject specific details of the mapping from raw fleshpoint coordinates to this component were too complex to be consistent with the PARAFAC framework. The final model explained over 90% of the variance and gave a succinct and physiologically plausible articulatory representation of the German vowel space. PMID- 10462808 TI - Development of adaptive phonetic gestures in children: evidence from vowel devoicing in two different dialects of Japanese. AB - High vowels between voiceless consonants are often devoiced in many languages, as well as in many dialects of Japanese. This phenomenon can be hypothesized to be a consequence of the adaptive organization of the laryngeal gestures to various conditions, including dialectal requirements. If this theory is correct, it may be possible to predict developmental changes in vowel devoicing based on the developmental improvement in the dialect-specific organization of the laryngeal gestures. To test this expectation, the developmental properties of vowel devoicing were investigated for 72 children of 4 and 5 years of age, and 37 adults in two dialects of Japanese. One was the Osaka dialect, with a low devoicing rate, and the other the Tokyo dialect, with a high devoicing rate. In the Tokyo dialect, the devoicing rate of children significantly increased and reached an adultlike level by the age of 5 years, whereas it remained low irrespective of age in Osaka. The vowel devoicing of 5-year-old children exhibited the same characteristics as that of the adults of their respective dialect. These results suggest that children growing up with the Tokyo dialect acquire the articulatory gestures which do not inhibit vowel devoicing by the age of 5 years, whereas children growing up with the Osaka dialect acquire those which inhibit the devoicing of vowels by the same age. The results fit in well with the predictions of the gestural account of vowel devoicing. It is also suggested that learning dialect-specific adaptive strategies to coordinate voicing and devoicing gestures as required to attain an adultlike vowel devoicing pattern is a long process: By the age of 5 years children have completed enough of this process to become members of their dialectal community. PMID- 10462809 TI - Speech segment durations and quantity in Icelandic. AB - Icelandic has a phonologic contrast of quantity, distinguishing long and short vowels and consonants. Perceptual studies have shown that a major cue for quantity in perception is relational, involving the vowel-to-rhyme ratio. This cue is approximately invariant under transformations of rate, thus yielding a higher-order invariant for the perception of quantity in Icelandic. Recently it has, however, been shown that vowel spectra can also influence the perception of quantity. This holds for vowels which have different spectra in their long and short varieties. This finding raises the question of whether the durational contrast is less well articulated in those cases where vowel spectra provide another cue for quantity. To test this possibility, production measurements were carried out on vowels and consonants in words which were spoken by a number of speakers at different utterance rates in two experiments. A simple neural network was then trained on the production measurements. Using the network to classify the training stimuli shows that the durational distinctions between long and short phonemes are as clearly articulated whether or not there is a secondary, spectral, cue to quantity. PMID- 10462810 TI - Acoustic correlates of talker sex and individual talker identity are present in a short vowel segment produced in running speech. AB - Although listeners routinely perceive both the sex and individual identity of talkers from their speech, explanations of these abilities are incomplete. Here, variation in vocal production-related anatomy was assumed to affect vowel acoustics thought to be critical for indexical cueing. Integrating this approach with source-filter theory, patterns of acoustic parameters that should represent sex and identity were identified. Due to sexual dimorphism, the combination of fundamental frequency (F0, reflecting larynx size) and vocal tract length cues (VTL, reflecting body size) was predicted to provide the strongest acoustic correlates of talker sex. Acoustic measures associated with presumed variations in supralaryngeal vocal tract-related anatomy occurring within sex were expected to be prominent in individual talker identity. These predictions were supported by results of analyses of 2500 tokens of the /epsilon/ phoneme, extracted from the naturally produced speech of 125 subjects. Classification by talker sex was virtually perfect when F0 and VTL were used together, whereas talker classification depended primarily on the various acoustic parameters associated with vocal-tract filtering. PMID- 10462811 TI - Glottal characteristics of male speakers: acoustic correlates and comparison with female data. AB - Acoustic measurements believed to reflect glottal characteristics were made on recordings collected from 21 male speakers. The waveforms and spectra of three nonhigh vowels (/ae, lambda, epsilon/) were analyzed to obtain acoustic parameters related to first-formant bandwidth, open quotient, spectral tilt, and aspiration noise. Comparisons were made with previous results obtained for 22 female speakers [H. M. Hanson, J. Acoust. Soc. Am. 101, 466-481 (1997)]. While there is considerable overlap across gender, the male data show lower average values and less interspeaker variation for all measures. In particular, the amplitude of the first harmonic relative to that of the third formant is 9.6 dB lower for the male speakers than for the female speakers, suggesting that spectral tilt is an especially significant parameter for differentiating male and female speech. These findings are consistent with fiberscopic studies which have shown that males tend to have a more complete glottal closure, leading to less energy loss at the glottis and less spectral tilt. Observations of the speech waveforms and spectra suggest the presence of a second glottal excitation within a glottal period for some of the male speakers. Possible causes and acoustic consequences of these second excitations are discussed. PMID- 10462812 TI - Cortical auditory evoked potential correlates of categorical perception of voice onset time. AB - The goal of this study was to examine the neural encoding of voice-onset time distinctions that indicate the phonetic categories /da/ and /ta/ for human listeners. Cortical Auditory Evoked Potentials (CAEP) were measured in conjunction with behavioral perception of a /da/-/ta/ continuum. Sixteen subjects participated in identification and discrimination experiments. A sharp category boundary was revealed between /da/ and /ta/ around the same location for all listeners. Subjects' discrimination of a VOT change of equal magnitude was significantly more accurate across the /da/-/ta/ categories than within the /ta/ category. Neurophysiologic correlates of VOT encoding were investigated using the N1 CAEP which reflects sensory encoding of stimulus features and the MMN CAEP which reflects sensory discrimination. The MMN elicited by the across-category pair was larger and more robust than the MMN which occurred in response to the within-category pair. Distinct changes in N1 morphology were related to VOT encoding. For stimuli that were behaviorally identified as /da/, a single negativity (N1) was apparent; however, for stimuli identified as /ta/, two distinct negativities (N1 and N1') were apparent. Thus the enhanced MMN responses and the morphological discontinuity in N1 morphology observed in the region of the /da/-/ta/ phonetic boundary appear to provide neurophysiologic correlates of categorical perception for VOT. PMID- 10462813 TI - Acquisition of a tactile-alone vocabulary by normally hearing users of the Tickle Talker. AB - Tactile-alone word recognition training was provided to six normally hearing users of the Tickle Talker, an electrotactile speech perception device. A mean group tactile-alone vocabulary of 31 words was learned in 12 h of training. These results were comparable to, or superior to, those reported for other tactile devices and Tadoma. With increased training the group became faster at learning tactually new words, which were introduced in small training sets. However, as their tactile-alone vocabulary grew, subjects required more training time to reach the pass criterion when evaluated on their recognition of their whole vocabulary list. A maximum possible vocabulary size was not established. The application of tactile-alone training with hearing-impaired users of the device is discussed. PMID- 10462814 TI - Vocal tract normalization for midsagittal articulatory recovery with analysis-by synthesis. AB - A method is presented that accounts for differences in the acoustics of vowel production caused by human talkers' vocal-tract anatomies and postural settings. Such a method is needed by an analysis-by-synthesis procedure designed to recover midsagittal articulatory movement from speech acoustics because the procedure employs an articulatory model as an internal model. The normalization procedure involves the adjustment of parameters of the articulatory model that are not of interest for the midsagittal movement recovery procedure. These parameters are adjusted so that acoustic signals produced by the human and the articulatory model match as closely as possible over an initial set of pairs of corresponding human and model midsagittal shapes. Further, these initial midsagittal shape correspondence need to be generalized so that all midsagittal shapes of the human can be obtained from midsagittal shapes of the model. Once these procedures are complete, the midsagittal articulatory movement recovery algorithm can be used to derive model articulatory trajectories that, subsequently, can be transformed into human articulatory trajectories. In this paper the proposed normalization procedure is outlined and the results of experiments with data from two talkers contained in the X-ray Microbeam Speech Production Database are presented. It was found to be possible to characterize these vocal tracts during vowel production with the proposed procedure and to generalize the initial midsagittal correspondences over a set of vowels to other vowels. The procedure was also found to aid in midsagittal articulatory movement recovery from speech acoustics in a vowel-to-vowel production for the two subjects. PMID- 10462815 TI - Nonlinear propagation effects on broadband attenuation measurements and its implications for ultrasonic tissue characterization. AB - A study is presented in which the influence of the pressure amplitude of the incident pulse on the estimated frequency dependency of the attenuation coefficient is shown. First, the effect is demonstrated with a simple theoretical model for both transmission and reflection measurements. Simulations show that for both measurement techniques a high-amplitude incident pulse results in a biased estimate of the attenuation coefficient due to nonlinear interaction of the different frequency components of the incident pulse. It is shown that in transmission and reflection measurements the biases have opposite signs. The effect of bandwidth, central frequency, and phase of the incident pulse on this bias is investigated. Second, the effect is demonstrated both in vitro, using a broadband through-transmission substitution technique on a tissue mimicking gelatine phantom, and in vivo, using reflection measurements with standard clinical equipment. The experimental results agree well with the theoretical model. The relevance of this study for ultrasonic tissue characterization is shown. PMID- 10462816 TI - Killer whale (Orcinus orca) hearing: auditory brainstem response and behavioral audiograms. AB - Killer whale (Orcinus orca) audiograms were measured using behavioral responses and auditory evoked potentials (AEPs) from two trained adult females. The mean auditory brainstem response (ABR) audiogram to tones between 1 and 100 kHz was 12 dB (re 1 mu Pa) less sensitive than behavioral audiograms from the same individuals (+/- 8 dB). The ABR and behavioral audiogram curves had shapes that were generally consistent and had the best threshold agreement (5 dB) in the most sensitive range 18-42 kHz, and the least (22 dB) at higher frequencies 60-100 kHz. The most sensitive frequency in the mean Orcinus audiogram was 20 kHz (36 dB), a frequency lower than many other odontocetes, but one that matches peak spectral energy reported for wild killer whale echolocation clicks. A previously reported audiogram of a male Orcinus had greatest sensitivity in this range (15 kHz, approximately 35 dB). Both whales reliably responded to 100-kHz tones (95 dB), and one whale to a 120-kHz tone, a variation from an earlier reported high frequency limit of 32 kHz for a male Orcinus. Despite smaller amplitude ABRs than smaller delphinids, the results demonstrated that ABR audiometry can provide a useful suprathreshold estimate of hearing range in toothed whales. PMID- 10462817 TI - Underwater temporary threshold shift induced by octave-band noise in three species of pinniped. AB - Pure-tone sound detection thresholds were obtained in water for one harbor seal (Phoca vitulina), two California sea lions (Zalophus californianus), and one northern elephant seal (Mirounga angustirostris) before and immediately following exposure to octave-band noise. Additional thresholds were obtained following a 24 h recovery period. Test frequencies ranged from 100 Hz to 2000 Hz and octave-band exposure levels were approximately 60-75 dB SL (sensation level at center frequency). Each subject was trained to dive into a noise field and remain stationed underwater during a noise-exposure period that lasted a total of 20-22 min. Following exposure, three of the subjects showed threshold shifts averaging 4.8 dB (Phoca), 4.9 dB (Zalophus), and 4.6 dB (Mirounga). Recovery to baseline threshold levels was observed in test sessions conducted within 24 h of noise exposure. Control sessions in which the subjects completed a simulated noise exposure produced shifts that were significantly smaller than those observed following noise exposure. These results indicate that noise of moderate intensity and duration is sufficient to induce TTS under water in these pinniped species. PMID- 10462818 TI - Comparison of electrohydraulic lithotripters with rigid and pressure-release ellipsoidal reflectors. II. Cavitation fields. AB - Dramatically different cavitation was produced by two separate acoustic pulses that had different shapes but similar duration, frequency content, and peak positive and negative pressure. Both pulses were produced by a Dornier HM-3 style lithotripter: one pulse when the ellipsoidal reflector was rigid, the other when the reflector was pressure release. The cavitation, or bubble action, generated by the conventional rigid-reflector pulse was nearly 50 times longer lived and 3 13 times stronger than that produced by the pressure-release-reflector pulse. Cavitation durations measured by passive acoustic detection and high-speed video agreed with calculations based on the Gilmore equation. Cavitation intensity, or destructive potential, was judged (1) experimentally by the size of pits in aluminum foil detectors and (2) numerically by the calculated amplitude of the shock wave emitted by a collapsing bubble. The results indicate that the trailing positive spike in the pressure-release-reflector waveform stifles bubble growth and mitigates the collapse, whereas the trough after the positive spike in the rigid-reflector waveform triggers inertially driven growth and collapse. The two reflectors therefore provide a tool to compare effects in weakly and strongly cavitating fields and thereby help assess cavitation's role in lithotripsy. PMID- 10462819 TI - Uncertainties in estimates of lesion detectability in diagnostic ultrasound. AB - Statistical properties of estimates of focal lesion detectability for medical ultrasonic imaging systems are investigated. Analytic forms for bias and variance of estimates of detectability of a lesion consisting of fully developed speckle embedded within a speckle background are derived. Bias and variance of estimates of detectability are investigated using a computer simulation and experiments on tissue-mimicking phantoms. This work offers a systematic methodology for interpreting measurements on phantoms in order to assess lesion detectability. In addition, it provides useful results which may be used to improve design of phantoms and experiments for imaging-system performance assessment. PMID- 10462820 TI - Love-theoretical analysis of periodic system of rods. PMID- 10462821 TI - The fine structure of the recovering auditory detection threshold. AB - Detection thresholds were gathered for a 2 kHz Gaussian-shaped probe (standard deviation = 0.5 ms), centered at intervals of as little as half a millisecond over 0-30 ms following a 200 ms, 97 dB SPL, 2 kHz tone. Surprisingly, there were small, sudden rises and falls superimposed on each subject's generally smooth recovery. Even more obvious were nonmonotonicities in the standard deviation of the cumulative normal fitted to each threshold's psychometric function. PMID- 10462822 TI - Phoneme recognition by cochlear implant users as a function of signal-to-noise ratio and nonlinear amplitude mapping. PMID- 10462823 TI - A prospective one-year follow-up study of patients with bipolar affective disorder. AB - BACKGROUND: Patients with affective disorder often have a poorer than expected prognosis for functional outcome between episodes. Previously, these patients were thought to have a better outcome between episodes, particularly Chinese patients. The aim of our study was to prospectively monitor symptoms, and vocational and residential functions of a group of patients with bipolar affective disorder one year after hospitalization. This study compare these data with index, admission data and discuss the relationship between patients' symptoms and functional outcome. METHODS: The Brief Psychiatric Rating Scale (BPRS), Modified Manic State Rating Scale (MMS) and Hamilton Depression Rating Scale (HDRS-21 items) were used to measure the patients' general psychiatric condition, manic and depressive symptoms, respectively. A modified Vocational State Index and modified Location Code Index Scale were used to determine levels of functioning. RESULTS: One-year follow-up measurements indicated that 75% of patients were free of symptoms or had mild psychiatric symptoms and 95% of patients were asymptomatic or mildly symptomatic for affective-manic condition. There was significant improvement in symptom outcome. However, only 46% of patients were employed at the one-year follow-up assessment, and as few as 12% worked at their expected level of employment, while 42% were rated as being unable to work and remained at an incapacitated occupational function. There was a significant tendency for increased independent living at one year of follow-up (74.6% vs 45.8%). More than 40% of patients were solely responsible for themselves, while over 50% required others to be responsible for them in terms of daily living. Additionally, 70% of first-admission subjects were employed at some level at one-year follow-up assessment, compared with only 31.8% of subjects with multiple admissions; 70% of first-admission patients were living independently compared with only 40% of subjects with multiple admissions. CONCLUSIONS: The results showed that the symptom outcome was superior to the functional outcome. These findings indicate that variable factors other than psychotic symptoms are related to the functional outcome in Chinese patients with bipolar affective disorder. Although treatment reduced the symptom intensity of affective episodes, a long-range rehabilitative program and strategy for prevention relapse is required to help functional outcome catch up with symptomatic amelioration. PMID- 10462824 TI - Circulating intercellular adhesion molecule-1 in chronic liver disease and hepatocellular carcinoma. AB - BACKGROUND: Intercellular adhesion molecule-1 (ICAM-1) is important in the migration and adhesion of immune cells from the circulation to their targets. A circulating form of ICAM-1 (cICAM-1) is elevated in the serum of patients with hepatitis and various cancers. Our aim was to determine the clinical significance of cICAM-1 in chronic liver diseases and hepatocellular carcinoma (HCC). METHODS: We measured the serum cICAM-1 levels in 91 patients with HCC, 47 with liver cirrhosis, 41 with chronic viral hepatitis and 32 healthy controls using an enzyme-linked immunosorbent assay. Ninety-eight patients had serial follow-up. RESULTS: The cICAM-1 levels in patients with HCC (737 +/- 212 ng/ml) and liver cirrhosis (593 +/- 145 ng/ml) were significantly higher than those in patients with chronic viral hepatitis (488 +/- 127 ng/ml) and controls (318 +/- 64 ng/ml). HCC patients had higher cICAM-1 levels than all the other groups. cICAM-1 levels in patients with chronic viral hepatitis correlated with serum alanine aminotransferase, prothrombin time and indocyanine green retention ratio. In HCC patients, alpha-fetoprotein levels and tumor size paralleled cICAM-1 concentrations. Furthermore, HCC patients with distant metastases had higher cICAM-1 levels than those without distant metastases, and levels of cICAM-1 in patients with stage IV HCC were higher than those in HCC patients with stages I, II or III disease. During a seven-year follow-up, 14 patients with chronic hepatitis developed liver cirrhosis when their cICAM-1 levels increased significantly. In 47 HCC patients, cICAM-1 levels decreased after tumor resection or transcatheter arterial embolization, and increased when the tumor recurred. cICAM-1 levels increased gradually in 14 patients with advanced HCC without specific therapy. CONCLUSIONS: CICAM-1 levels are increased in patients with chronic hepatitis, liver cirrhosis and HCC. This reflects the severity and progression of chronic liver disease and HCC. Sequential measurements of cICAM-1 seem more valuable than a single-point assessment in evaluating the progress of liver disease. PMID- 10462825 TI - Relationship between pterygium/pinguecula and sunlight exposure among postmen in central Taiwan. AB - BACKGROUND: The purpose of this study was to investigate outdoor hazards and their relationship to conjunctival disorders experienced by postmen. METHODS: Three hundred and ninety-four employees (248 postmen and 146 officers) working in 11 post offices in central Taiwan were recruited and participated in the 1994 annual labor health examination. Pinguecula and pterygium were diagnosed by an ophthalmologist. Meanwhile, detailed personal and occupational information was obtained using a structured questionnaire. The cumulative occupational sunlight exposure was calculated for each postman by considering the duration of their employment as postmen, the average working hours per day and their spectacle use (sunglasses and eyeglasses). A logistic regression was used to analyze the relationship between cumulative occupational sunlight exposure and pinguecula. RESULTS: Among the postmen, the prevalences of pinguecula and pterygium were 62.9% and 7.3%, respectively. The outdoor nature of postal work was significantly associated with the occurrence of pinguecula as well as pterygium (p < 0.05). When the cumulative occupational sunlight exposure increased by one unit (one year x hour/day), the risks of developing pinguecula and pterygium were raised by 2.1% and 0.8%, respectively. CONCLUSIONS: The results indicate that conjunctival disorders were associated with the cumulative occupational sunlight exposure of postmen working outdoors. This study reinforces the importance of ocular protection from sunlight. PMID- 10462826 TI - Safety of distilled water as an irrigating fluid for transurethral resection of the prostate. AB - BACKGROUND: Irrigating fluid is necessary for transurethral resection of the prostate (TURP). Initially, distilled water was used as irrigating fluid and some advantages and complications were noted. Although other solutions were used to substitute for distilled water, we collected data for changes in electrolytes and hematocrit (Hct) during surgery to evaluate the safety of distilled water as irrigating fluid for TURP with suprapubic trocar cystostomy. METHODS: Fifty-three patients who underwent TURP using distilled water as irrigating fluid were investigated. TURP was performed with suprapubic trocar cystostomy routinely. Preoperative and postoperative blood samples were collected and analyzed for electrolytes and Hct to study any changes during surgery. RESULTS: The change in plasma sodium concentration was not statistically significant, but the decreases in blood Hct and plasma potassium concentration were statistically significant. There was no major operative complication and no clinical-evident related postoperative renal impairment. CONCLUSIONS: Distilled water is a relatively safe solution as an irrigating fluid for TURP under drainage with suprapubic trocar cystostomy, but attention must be paid to the inevitable blood loss during surgery. PMID- 10462827 TI - Dead-on-arrival patients in Panchiao, Taipei. AB - BACKGROUND: The present emergency medical service (EMS) system in the Panchiao area includes operations in Panchiao, Sulin, Sunshia, Chung Ho and Touchen, which is in the early developmental stage. Educational programs such as emergency medical technician (EMT)-I, and -II have been provided for less than 25% of firefighters in that area. Several mass educational programs in cardiopulmonary resuscitation (CPR) were also provided for the inhabitants of the area. The purpose of this study was to establish a database of dead-on-arrival (DOA) patients during this early stage of the EMS system development so that comparisons of EMS performance could be assessed to identify any future needs of the EMS system in the Panchiao area. METHODS: A patient was verified as DOA if there was no pulse nor a response to stimulation. For every DOA patient, several parameters were recorded in a registration book, which began January 1, 1995, and included the following: name, gender, age, mode of transportation, time to hospitalization, on-scene CPR, prehospital basic life support, possible etiology, response to advanced cardiac life support (ACLS), disposition and outcome. The registration book was reviewed and a follow-up study was designed, which covered the entire year of 1995. RESULTS: There were 292 DOA patients brought to the emergency room of the Far Eastern Memorial Hospital in Panchiao, Taipei, between January 1, 1995, and December 31, 1995. The study population comprised 190 (65.1%) men and 102 (34.9%) women. The mean age of the men was 51.8 +/- 22.3 years (mean +/- standard deviation). The mean age of the women was 59.8 +/- 24.4 years. Emergency service ambulances transported most DOA patients, though a few were transported by private ambulance from nursing homes. There were only two incidents in which prehospital CPR was conducted. The time to hospitalization varied from five to 170 minutes and averaged 38.3 minutes. Following ACLS, 244 (83.6%) patients were pronounced dead in the emergency room. Cardiovascular and respiratory problems were the leading etiologies and trauma was the second. There were 48 patients (16.4%) who responded to ACLS, but only eight (2.7%) survived until hospital discharge. CONCLUSIONS: Delayed initiation of the EMS system resulted in an average time to hospitalization of 38.3 minutes. Cardiovascular collapse was the leading etiology in DOA patients. There were only two patients who received prehospital CPR, suggesting that layperson CPR and EMT education cannot be overemphasized. PMID- 10462828 TI - Management of stage pTxN+ adenocarcinoma of the prostate: influence of radical prostatectomy on progression-free interval. AB - BACKGROUND: Whether stage pN+ prostate cancer patients will benefit from radical prostatectomy remains a controversial issue. We assess the impact of radical prostatectomy on a progression-free interval for patients with stage pN+ prostate cancer. METHODS: From October 1990 to June 1997, 25 patients were diagnosed with stage pTxN+ adenocarcinoma of the prostate in our institute. Ten of these patients were treated with radical retropubic prostatectomy (RRP) due to false negative frozen sections at staging pelvic lymphadenectomy. In 15 patients, radical surgery was discontinued because of grossly enlarged lymph nodes and/or positive frozen section results during staging pelvic lymph node dissection (PLND). All patients received immediate androgen ablation therapy with or without radiotherapy. Serum prostate-specific antigen (PSA) was regularly monitored in all patients. RESULTS: During a median follow-up period of 33 months (range, 15 89 months), eight patients (53%) in the PLND group and two (20%) in the RRP group experienced disease progression as defined by significant elevation of serum PSA concentrations. The median progression-free intervals were 46 and 51 months, respectively. Both univariate and multivariate analyses failed to show statistically significant differences in the progression-free survival between the RRP and PLND groups. CONCLUSIONS: Based on our limited experience, surgical removal of the primary tumor may have only a marginal effect in terms of the progression-free interval for prostatic cancer patients with metastasized pelvic lymph nodes (p = 0.124, log-rank test). The follow-up period was not long enough to evaluate the difference in cancer-specific survival. PMID- 10462829 TI - Intracranial pathology: comparison of intraoperative ultrasonography with computerized tomography and magnetic resonance imaging. AB - BACKGROUND: High-resolution, real time, intraoperative ultrasonography (IOUS) has been used in a number of cases in our neurosurgical department to identify, localize and characterize intracranial abnormalities. We compare the images of IOUS with those of computerized tomography (CT) and magnetic resonance imaging (MRI) in order to enhance our experience and confidence. METHODS: IOUS images were compared with preoperative CT scans and/or MRIs in 30 intracranial masses. Tumor size and margin, echogenicity and edema were the features compared. RESULTS: The images of IOUS, CT and MRI correlated closely except for images of primary gliomas. IOUS was more effective than CT and MRI in delineating the extent of a glioma, determining whether a lesion was cystic, with or without septation, or solid and in guiding surgical procedures. The contrast enhancement available with CT and MRI could not be demonstrated with IOUS. CONCLUSIONS: IOUS provides exquisite detail of intracranial pathology as well as, or even better than, CT and MRI. Its application should be emphasized to minimize damage of normal brain tissue and increase the completeness of tumor resection. PMID- 10462830 TI - Autologous peripheral blood stem cell transplantation using simplified procedures for collection and freezing. AB - BACKGROUND: High-dose therapy with peripheral blood stem cell (PBSC) transplantation has been used increasingly for chemosensitive malignancies. The conventional procedures for collection and freezing of PBSCs are time-consuming and expensive. The optimal conditions for priming, collection and freezing of PBSCs have yet to be simplified. METHODS: Simplified procedures for mobilization, collection and freezing were developed to provide PBSC transplantation. Twenty two cancer patients were given intensive chemotherapy with a variety of regimens using granulocyte-colony stimulating factor (G-CSF, filgrastim) 300 micrograms/d subcutaneously to mobilize PBSCs. A rapidly rebounding white blood cell (WBC) count from nadir was used to predict the time of peak PBSC release and plan leukapheresis. Leukapheresis was performed, when the WBC count was greater than 1.0 x 10(9)/l. RESULTS: Leukapheresis was performed in all 22 patients on day 10 to 14 after completing intensive chemotherapy. G-CSF to mobilize PBSCs was administered for a median of 10 days (range, 7-12 days). Twenty-one of 22 patients achieved the target yield of greater than 2 x 10(6) CD34+ cells/kg of body weight (BW). This was achieved in a median of two harvests (range, 2-4), with a median processed blood volume of 10.2 l/apheresis. A median of 6.4 x 10(8) mononuclear cells/kg BW and 5.2 x 10(6) CD34+ cells/kg BW for each patient were obtained. After high-dose cancer chemotherapy and PBSC transplantation, rapid and sustained hematopoietic engraftment occurred in all but one patient who had an inadequate number of CD34+ cells. In patients with an adequate target yield of CD34+ cells, the median times to achieve an absolute neutrophil count of greater than 0.5 x 10(9)/l and a platelet count of greater than 20 x 10(9)/l were nine days (range, 7-12 days) and 12 days (range, 8-14 days), respectively. CONCLUSIONS: Adequate PBSCs can be achieved within a median of two aphereses in most eligible cancer patients. Apheresis should be performed on day 10 to 14 after completing intensive chemotherapy, followed by G-CSF 300 micrograms/d. This simple and effective approach makes PBSC transplantation more practicable for a wider range of malignant diseases. PMID- 10462831 TI - Retrospective analysis of 196 patients with suspected pulmonary thromboembolism. AB - BACKGROUND: There has not been a statistical analysis reporting on patients with pulmonary thromboembolism (PE) in Taiwan. To identify the most common manifestations in patients with PE in Taiwan, we conducted this retrospective study. METHODS: We collected and analyzed the medical records of 196 patients (53 men, mean age, 60 years; 143 women, mean age, 46 years) whose clinical symptoms and signs, chest radiographs and lung scan findings were suggestive of PE. Conventional chest radiography was performed 24 hours to 48 hours before lung scans. The radiopharmaceuticals used in lung scans were 133Xenon for ventilation studies and 99mTechnetium macroaggregated albumin for perfusion studies. All patients had received anticoagulant therapy (heparin and coumadin) to improve PE. RESULTS: The two most common symptoms and signs of PE in the 196 patients were dyspnea (172/196, 87.8%) and tachypnea (176/196, 89.8%). The most common abnormal finding on chest radiography was increased lung markings, which were present in about 36.7% (72/196) of patients. Lung scans showed two or more, large, mismatched, segmental defects (high probability of PE) in both lungs in about 88.8% (174/196) of patients. After anticoagulant therapy, PE, as seen on lung scans, was resolved within four weeks in approximately 93.9% (184/196) of patients. CONCLUSIONS: In Taiwan, a woman aged 40 to 50 years, with no prior history of other medical problems or surgical procedures, with sudden onset of dyspnea or tachypnea, normal or increased lung markings on chest radiography and two or more large mismatched segmental defects on lung scans, is at greatest risk of developing PE. Most patients with PE showed a good response to anticoagulant therapy within four weeks. PMID- 10462832 TI - Transcranial Doppler monitoring with head-upright tilting in patients with syncope. AB - BACKGROUND: The aim of our study was to evaluate the effects of orthostatic stress produced by the head-upright tilt test on human cerebral hemodynamics by transcranial Doppler sonography. METHODS: We studied 60 subjects who were divided into two groups; one of normal controls (n = 43) and one of patients suffering from syncope (n = 17). A 30-minute head-upright tilt test was conducted on all subjects, and heart rate and blood pressure were monitored by surface electrocardiography and cuff sphygmomanometry, respectively. Cerebral blood flow velocity and cerebral vasoreactivity were continuously monitored using transcranial Doppler sonography. RESULTS: The maximal decreases in mean blood pressure of controls and patients with syncope were 2.6 +/- 7.8% and 0.5 +/- 7.9% of baseline, respectively. The maximal decreases in mean blood flow velocity in the middle cerebral artery between the two groups reached 19.6 +/- 6.2% and 30.7 +/- 14.1% of baseline, respectively (p < 0.05). The increases in pulsatility index between the two groups were 15.4 +/- 14.3% and 16.9 +/- 21.1% of baseline, respectively. CONCLUSION: The responses of cerebral blood flow to upright tilting differed significantly between normal controls and patients with syncope, implying that the latter may suffer an impairment of cerebral autoregulation. Further studies are needed to ascertain what clinical implications this finding might have. PMID- 10462833 TI - Urethral diverticulum presenting with chronic pelvic pain: a case report. AB - Urethral diverticula are rarely encountered in the gynecologic out-patient setting. However, this condition probably occurs more frequently than it is diagnosed. The patient frequently notes signs of lower urinary tract irritation. Urinary dribbling accompanied with dyspareunia and dysuria constitutes a classic triad for urethral diverticula. Symptoms of chronic pelvic pain only occur in a minority of patients. We report a case of urethral diverticulum presenting chiefly with chronic pelvic pain. The patient underwent multiple investigative operations before a correct diagnosis was made. When confronted with a patient presenting with chronic pelvic pain, a gynecologist should retain a high index of suspicion for a urethral diverticulum in addition to other gynecologic conditions. PMID- 10462834 TI - Octreotide effect on hypersecretion of growth hormone in a patient with fibrous dysplasia: a case report. AB - We report the case of a 17-year-old adolescent boy with polyostotic fibrous dysplasia and hypersecretion of growth hormone (GH). The fasting serum GH, insulin-like growth factor-I (IGF-I), alkaline phosphatase activity and osteocalcin levels were all elevated. The GH secretion was stimulated by thyrotropin-releasing hormone and was not suppressed by an oral glucose test. Magnetic resonance imaging of the sella turcica showed no abnormal findings. The patient was treated with octreotide, 100 micrograms subcutaneous injection three times a day for two weeks to observe the effects of octreotide on growth hormone secretion. GH and IGF-I secretions were suppressed by octreotide therapy, while alkaline phosphatase activity and osteocalcin secretion were partially suppressed. We suggest that the high bone turnover states in this patient may be attributed to both hypersecretion of growth hormone and the polyostotic fibrous dysplasia itself. PMID- 10462835 TI - Lymphatic cyst of the colon: a case report. AB - A rare case of a lymphatic cyst of the ascending colon is reported. The lobulated and fluctuant lesion, located in the hepatic flexure, was diagnosed by barium enema and colonoscopy, incidentally. Surgical treatment with segmental resection of the ascending colon was done because of the risk of obstruction and the question of an underlying malignancy. The clinical features, appropriate treatment, preoperative diagnosis and histopathology are discussed. We emphasize that lymphatic cysts are very rare and are difficult to identify by radiology or colonoscopy before surgery. They must be included in the differential diagnosis of submucosal tumors such as lipomas, leiomyomas and hemangiomas. PMID- 10462836 TI - Cervical necrotizing fasciitis: a case report. AB - Necrotizing fasciitis is a severe soft tissue infection characterized by cutaneous necrosis, suppurative fasciitis, vascular thrombosis and extreme systemic toxicity. Involvement of head and neck structures is rare, but occur most frequently in patients with diabetes and chronic alcoholism. Once initiated, the disease progresses rapidly and diffusely, involving adjacent fascial spaces. Necrotizing fasciitis may also extend to the cervical viscera, mediastinum and anterior chest wall. A 65-year-old chronic alcoholic man, with long-standing diabetes and liver cirrhosis under irregular treatment is described. The patient developed a deep neck infection from a buccal abscess after a local incision. The infection then extended to an orocutaneous fistula and deep neck superficial and middle layer fascias, with necrotizing fasciitis. Management requires early recognition, high doses of appropriate antimicrobial therapy, early surgical drainage and radical debridement of necrotic tissue. The disease carries a high rate of morbidity and mortality, especially in the elderly. PMID- 10462837 TI - [Factors that modify de- and remineralization in dental enamel from the aspect of caries susceptibility]. AB - The course of caries depends on an equilibrium between de- and remineralization of enamel. Epidemiologic studies performed during recent years in Poland have demonstrated a high incidence and severity of caries in children and teenagers. Therefore, the actual program of caries prevention covering the whole population of children and teenagers has not performed to the expectations, possibly because the program protocol does not discriminate between children susceptible and resistant to caries. The aims of this work included: 1. Analysis of tooth groups and surfaces as to their susceptibility to caries; 2. Evaluation of the influence of de- and remineralization modifying factors such as: a) dental plaque and sugars, b) caries-causing bacteria, c) enamel resistance to acids, d) biochemical properties of saliva--on the susceptibility to caries; 3. Evaluation of the uptake of fluorides from chewing gum by enamel in relationship to caries susceptibility. An initial clinical examination of permanent teeth was performed in 292 children aged 12 years. Subsequently, two groups were formed: one with 45 children free from caries and another with 77 children at high risk of caries. Control examinations of permanent teeth in both groups were carried out after one and three years from the initial examination. The clinical examination included a check-up of secondary dentition, counting of erupted teeth, sealed teeth, evaluation of oral hygiene status, gingivae, abnormalities in mineralization and measurement of the rate of microdemineralization. The biochemical analysis of saliva was done to measure the content of fluorides, calcium, phosphorus and magnesium, as well as pH. Microsamples of enamel were collected using acid biopsy and the content of calcium and fluorine in the superficial and underlying layer was assayed. The thickness of both layers obtained by biopsy was also measured. Commercial test kits--Dentocult SM and Dentocult LB, served to determine the amount of caries-causing bacteria in saliva. Additionally, the children and their mothers responded to a questionnaire on the diet, previous diseases, therapies with antibiotics and health habits. The results were analysed using Statgraphics software. Differences between means were checked using Student t-test in its classical form or as modified by B. L. Welch. Linear correlation indices were calculated and Spearman rank correlation test was performed. On the basis of the results obtained the following conclusions were drawn: 1. 12-year old children at high risk of caries need intense dental care due to: a) extensive involvement of masticatory surfaces of molar teeth and contact surfaces of premolar teeth, b) treatment inappropriate to actual needs in this group of children. 2. Effective programs to improve the knowledge and health of children at high risk to caries should be introduced due to the following facts: a) oral hygiene was unsatisfactory in the examined children, b) children consumed large quantities of sweets. 3. Disturbances in the mineralization of enamel and the use of antibiotics in childhood were without influence on the susceptibility of teeth to caries. 4. The content of fluorides, calcium and phosphorus in the saliva of children susceptible to caries could have been below the level necessary to maintain an equilibrium between de- and remineralization of enamel. 5. Susceptibility of teeth to caries could be due to differences in the chemical composition and structure of enamel, as it appears from the content of fluorides and calcium, depth of enamel biopsy and uptake of fluorides from chewing gum. 6. Chewing of fluoride-containing gums is a supportive measure in caries prevention, particularly recommended in children at high risk of caries. 7. Caries prevention should be individualized and matched to caries risk. PMID- 10462838 TI - Physiological responses to maximal treadmill and deep water running in the young and the middle aged males. AB - In this study we investigated the effects of age factors on physiological responses to deep water running (DWR) compared with those of treadmill running (TMR) while the water and ambient temperatures were kept in thermoneutral conditions. Fourteen young healthy non-smoker males (Age = 20.4 +/- 3.3 years, Height = 170.7 +/- 6.2 cm, Weight = 65.1 +/- 11.4 kg) and fourteen middle aged healthy non-smoker males (Age = 38.6 +/- 4.4 years, Height = 171.8 +/- 4.7 cm, Weight = 75.4 +/- 9.6 kg) were selected for the study. Two maximal tests, one on the treadmill and the other running in deep water using the Wet Vest (Lincoln life jacket) were completed by each subject. The order of trial was counterbalanced with half of the subjects in each group completing TMR first and the rest of those completing DWR first. Although the young males had significantly (P < 0.05) higher relative VO2max, HRmax than the middle aged males, there were no significant differences in absolute VO2max, respiratory exchange ratio (RER), maximal ventilation (VEmax), ratings of perceived exhaustion (RPE), and peak blood lactate values between the two groups. In conclusion, the VO2max, HRmax, VEmax, and peak blood lactate value in response to DWR were significantly lower than those to TMR in both the young and the middle aged males in the thermoneutral conditions. However, there was no significant interaction between age and exercise modes other than RPE of legs at maximal efforts in the present study. We found that the decrease in the maximal physiological responses to DWR compared to TMR is not different between the young and middle aged males. PMID- 10462839 TI - Clothing selection behavior of the aged women for thermal comfort. AB - Wearing behavior and thermoregulatory responses of five young women (YG; 20 +/- 1 yr) and five aged women (AG; 65 +/- 3 yr) to indoor cold in summer were investigated in this study. The subjects were exposed to 21.0 +/- 0.5 degrees C and 55 +/- 5% RH while seated during a 90-minute experiment. The subjects were allowed to select and wear for thermal comfort clothing whenever they needed additional clothing during the experiment. Rectal temperature (Tre) and temperatures of 7 sites (head, chest, forearm, hand, thigh, leg, foot) of the skin of the subjects were measured every 10 minutes. Mean skin temperature (Tsk) of the subject was obtained every 10 minutes. First selection time of additional clothing was monitored and weight of selected total clothing was calculated. The results for this study were as follows: Tre and Tsk gradually decreased in YG and AG, however Tre decreased less than Tsk which decreased greater in AG than YG (p < 0.01). AG's first selection of additional clothing and thermal sensation response were slower than YG's. Furthermore, total clothing weight was less in AG than YG. It was concluded that clothing selection behavior would modify the intrinsic thermoregulatory responses of the aged women to the cold stress in the summer. PMID- 10462840 TI - Anthropometric and biomechanical characteristics on body segments of Koreans. AB - This paper documents the physical measurements of the Korean population in order to construct a data base for ergonomic design. The dimension, volume, density, mass, and center of mass of Koreans whose ages range from 7 to 49 were investigated. Sixty-five male subjects and sixty-nine female subjects participated. Eight body segments (head with neck, trunk, thigh, shank, foot, upper arm, forearm and hand) were directly measured with a Martin-type anthropometer, and the immersion method was adopted to measure the volume of body segments. After this, densities were computed by the density equations in Drillis and Contini (1966). The reaction board method was employed for the measurement of the center of mass. Obtained data were compared with the results in the literature. The results in this paper showed different features on body segment parameters comparing with the results in the literature. The constructed data base can be applied to statistical guideline for product design, workspace design, design of clothing and tools, furniture design and construction of biomechanical models for Korean. Also, they can be extended to the application areas for Mongolian. PMID- 10462841 TI - Individual differences in physiological responses and type A behavior pattern. AB - The relationships between individual differences in psychophysiological responses and tendency of Type A behavior pattern (TABP) were investigated during mental arithmetic (MA) at a steady rhythm, challenging calculation (Uchida-Kraepelin serial addition test: UK test), music listening, and exposure to an 80 dB SPL of white noise. Each mental task was sustained for 5 minutes. Sixteen healthy Japanese adults, (10 males and 6 females) with an age from 18 to 36 years old volunteered for this study. The KG's Daily Life Questionnaire (KG Questionnaire) was used to investigate the tendency toward TABP, which included three sub factors: aggression-hostility, hard-driving and time urgency, and speed-power items. Recorded physiological variables were respiratory rate (RR), skin resistance response (SRR), eyeblinks, and heart rate (HR) calculated using frequency analysis to render high frequency power (HF) and the ratio of low/high frequencies (L/H ratio). During the MA and UK tests, significant increases in HR, L/H ratio, RR occurred, while significant decreases in HF were observed. Eyeblinks significantly increased during the MA test and significantly decreased during the UK test. During music and white noise, no significant changes occurred except for SRR, which decreased significantly. The coefficient of variances in each response was over 20% for almost all variables, indicating that individual differences in the magnitude of each response were large, even if the direction (increase or decrease) of the change was the same in almost all subjects. The highest correlation coefficients (r) between the mean values of relative magnitude for each variable and TABP scores during the MA and UK tests were obtained for the L/H ratio (MA: r = 0.591, UK test: r = 0.577) and the RR (MA: r = -0.576, UK test: r = -0.511). These values were statistically significant (p < 0.05). Similar results were obtained for TABP sub-factors. Though other investigations have reported relationships between HF and TABP, we found no significant relationship. It was suggested that sympathetic nerve activity became greater for TABP individuals than for Type B individuals under stress conditions. PMID- 10462842 TI - Physiological effects of sudden change in illuminance during dark-adapted state. AB - To derive an optimal illuminance of nighttime illumination, we conducted an experiment with 7 healthy young individuals and 7 healthy elderly individuals as subjects. After 20 minutes of adaptation to darkness, subjects were exposed to illumination under 5 conditions comprising 0.5 lx, 1 lx, 3 lx, 10 lx, or 30 lx vertical illuminance of the facial region, and heart rate variability (HRV) and electroencephalogram (EEG) were measured, and discomfort was evaluated by subjective report. Results of LF/(LF + HF) (LF = low frequency, HF = high frequency) demonstrated a V-shaped trend for the young groups beginning during exposure and ending post exposure, with 3 lx conditions representing the minimum value, a value markedly lower than that for 30 lx conditions. From these results we inferred that approximately 3 lx illuminance could best suppress physiological stress. Evaluation of discomfort by subjective report also demonstrated an increase in discomfort evaluation scores under high illuminance conditions. The alpha-wave proportion of EEG during exposure fell markedly on 3 lx or higher illuminance conditions, and we inferred that visual sensory information and cortical activity level were adequately attained in 3 lx or higher illuminance conditions. These results suggest that the optimal illuminance of nighttime illumination is approximately 3 lx. PMID- 10462843 TI - Evaluation of microbiological contamination in a museum. PMID- 10462844 TI - Portable infrared analyzers. PMID- 10462845 TI - Smart searching on the World Wide Web. PMID- 10462846 TI - Control of dusts from sanding in auto body repair shops. National Institute for Occupational Safety and Health. PMID- 10462847 TI - An evaluation of the effectiveness of lead paint hazard reduction when conducted by homeowners and landlords. AB - This research project was conducted in collaboration with the Iowa Department of Public Health to evaluate whether property owners who follow recommended procedures for lead-based paint removal/repair can do the work safely and effectively. This study included 29 homes where a lead-based paint hazard had been identified and lead-based paint was removed or repaired (hazard reduction). Exposure evaluation included pre-project surface dust wipe sampling, air monitoring during lead-based paint removal, post-project surface dust wipe sampling, and pre- and post-project blood samples from adult study participants. The comparison of surface dust wipe samples taken before and after lead paint hazard reduction was used to evaluate the effectiveness of lead paint hazard reduction. The lead loadings on window sill surfaces in the work area were significantly lower after completion of the project (p = 0.04), and the lead based paint removal did not contaminate the adjoining living area. The proportion of homes with surface dust lead loading exceeding Department of Housing and Urban Development (HUD) clearance standard was 73 percent pre-project and 38 percent post-project. Personal airborne exposures during lead removal activities (geometric mean = 59.3 micrograms/m3) reinforce the need for respiratory protection and good hygiene. There was no difference in adult pre-/post-blood levels, indicating that participants did remove lead in a safe manner with respect to their own exposures. The results indicate that hazard reduction can be done effectively when recommended procedures for the removal of lead-based paint are followed. PMID- 10462848 TI - Exposure assessment of laboratory students. AB - The Massachusetts Institute of Technology (MIT) has two kinds of laboratories, teaching for undergraduate students and research laboratories for graduate students and research staff. The objective of this study is to determine chemical exposures during teaching and research activities. There are three hypotheses in this study: (1) Exposures in academic laboratories are well below health standards; (2) Students in undergraduate teaching laboratories have less chemical exposure compared to students in graduate research laboratories; and (3) Students in different disciplines are expected to have different exposures. From September 1996 to December 1996, 132 air samples were collected from both teaching and research laboratories in the departments of Material Sciences and Engineering, Chemical Engineering, and Biology. The most frequently sampled chemicals in these three departments were cobalt, styrene, and formaldehyde, respectively. A total of 23 different agents were measured. In this study, the American Conference of Governmental Industrial Hygienists (ACGIH) threshold limit value (TLV)-short-term exposure limit (STEL) is used as the health-effect standard for exposure time less than four hours. The ACGIH TLV-TWA (time-weighted average) is used as the standard for exposure times equal to or greater than four hours. The ratio of measured concentrations to the appropriate ACGIH standard was then calculated. The geometric mean of the ratio for the total samples was 0.34 percent of the standards. There were 70 samples from teaching laboratories (geometric mean = 0.38% of the standards), and 62 samples from research laboratories (geometric mean = 0.08% of the standards). The chemical exposures relative to the standards in teaching laboratories were statistically higher than in research laboratories (p-value < 0.001). Information about personal protective equipment and the use of laboratory chemical hoods was also collected. The differences in use of personal protective equipment (PPE) among these departments was not statistically significant. From the air sampling results, we concluded that (1) Chemical exposures in the academic laboratories in this study were all well below the health standards; (2) Undergraduate students in teaching laboratories had higher chemical exposures than graduate students in research laboratories; (3) Chemical exposures among departments were not significantly different; and (4) Hazard communication, safety training, and laboratory rules enforcement are important for protection and may be the reason that the results from this study indicate that chemical exposures in this academic institution are well below the health standards under normal operations. PMID- 10462849 TI - The effect of room size and general ventilation on the relationship between near and far-field concentrations. AB - Measurements of personal exposure level are generally greater than or equal to concentrations found at some fixed location in the body of the workroom. This article presents a theoretical analysis of such differences using a simple model of exposure, implemented as a two-compartment mass balance model with a constant emission rate. Simulated exposure levels (termed the near-field) and the concentration at a fixed location (the far-field) were obtained for five room sizes (30 to 3000 m3), each with five general ventilation rates (0.3 to 30 air changes per hour) and three levels of air movement in the vicinity of the worker (generally 3 to 30 m3/min). The ratio of near- to far-field concentrations from the simulations ranged from unity in small poorly ventilated rooms to 24 in large well ventilated areas, which compares favorably with actual measurement data. The predicted concentrations obtained in small rooms (< 100 m3) with less than one air-change per hour were almost 40 times higher than in larger rooms. The results from these simulations have been used to modify the original exposure model to better reflect the likely effect of general ventilation on workers' exposure. PMID- 10462850 TI - A Bayesian approach to retrospective exposure assessment. AB - A variety of health effects are caused by chronic, cumulative exposure over time to pollutants. In these cases, to establish dose-response relationships for epidemiological and risk assessment purposes, it is vital to determine the exposures of individuals or cohorts as functions of time. Most existing occupational exposure databases, however, do not contain continuous records of historical exposures to airborne contaminants. These gaps in the historical record may be filled by using the knowledge base that experts and professionals in the field possess. In this article we present a new framework, based on Bayesian probabilistic reasoning, for obtaining estimates of exposure histories for airborne particulates from limited historical measurements, using subjective expert judgment. The framework has great potential applications in instances where there is sparse information or missing data on past exposures. Expert judgment, in the form of inputs to physical models, provides additional knowledge to retrospectively estimate exposure as a function of time from discrete and incomplete measurements. The expert judgments are informed by knowledge of historical plant conditions and work practices, and models describing process dependent aerosol generation, ventilation, and worker activity patterns. The result will be probability distributions of the exposure of task-groups of workers as a function of time, in the form of a matrix. PMID- 10462851 TI - A method for detecting breakthrough of organic solvent vapors in a charcoal tube using semiconductor gas sensors. AB - This study developed a method for detecting organic vapors that break through charcoal tubes, using semiconductor gas sensors as a breakthrough detector of vapors. A glass column equipped with two sensors was inserted in Teflon tubing, and air containing organic vapor was introduced at a constant flow rate. After the output signal of the sensors became stable, a charcoal tube was inserted into the tubing at the upstream of the sensors. The resistance of the sensors was collected temporally in an integrated circuit (IC) card. The vapor concentration of the air near the sensors was measured with a gas chromatograph (GC) equipped with a flame ionization detector (FID) at intervals of 5 minutes to obtain the breakthrough curve. When the relative humidity was zero, the output signals of the sensors began to change before the breakthrough point (1% breakthrough time). This tendency was almost the same for methyl acetate, ethyl acetate, isopropyl alcohol (IPA), toluene, and chloroform. For dichloromethane and 1,1,1 trichloroethane, the time when the sensor output signals began to rise was almost the same as the breakthrough point. When the relative humidity was 80 percent, the sensors could also detect many vapors before the breakthrough point, but they could not perceive dichloromethane and chloroform vapors. A personal sampling system with a breakthrough detector was developed and its availability is discussed. PMID- 10462852 TI - Factory performance evaluations of engineering controls for asphalt paving equipment. AB - This article describes a unique analytical tool to assist the development and implementation of engineering controls for the asphalt paving industry. Through an agreement with the U.S. Department of Transportation, the National Asphalt Pavement Association (NAPA) requested that the National Institute for Occupational Safety and Health (NIOSH) assist U.S. manufacturers of asphalt paving equipment with the development and evaluation of engineering controls. The intended function of the controls was to capture and remove asphalt emissions generated during the paving process. NIOSH engineers developed a protocol to evaluate prototype engineering controls using qualitative smoke and quantitative tracer gas methods. Video recordings documented each prototype's ability to capture theatrical smoke under "managed" indoor conditions. Sulfur hexafluoride (SF6), released as a tracer gas, enabled quantification of the capture efficiency and exhaust flow rate for each prototype. During indoor evaluations, individual prototypes' capture efficiencies averaged from 7 percent to 100 percent. Outdoor evaluations resulted in average capture efficiencies ranging from 81 percent down to 1 percent as wind gusts disrupted the ability of the controls to capture the SF6. The tracer gas testing protocol successfully revealed deficiencies in prototype designs which otherwise may have gone undetected. It also showed that the combination of a good enclosure and higher exhaust ventilation rate provided the highest capture efficiency. Some manufacturers used the stationary evaluation results to compare performances among multiple hood designs. All the manufacturers identified areas where their prototype designs were susceptible to cross-draft interferences. These stationary performance evaluations proved to be a valuable method to identify strengths and weaknesses in individual designs and subsequently optimize those designs prior to expensive analytical field studies. PMID- 10462854 TI - Rural public health matters. PMID- 10462853 TI - Medicare is fundamental to the health care of Australia. PMID- 10462855 TI - Transitions and inequalities in acute myocardial infarction mortality in New South Wales, 1969-94. AB - OBJECTIVE: To chart the transition in myocardial infarction mortality in NSW between 1969 and 1994 and to determine whether spatial, marital and occupational status differentials increased over time, despite overall mortality decline. METHOD: The study was undertaken in NSW for the 25-year period from 1969 when mortality from ischaemic heart disease declined. The demographic and spatial analysis involved descriptive epidemiology and utilised the unit list mortality files of the Australian Bureau of Statistics which are based on detailed death certificate information. RESULTS: Spatial, marital and occupational status differentials in myocardial infarction mortality increased over time, more especially variations by occupational status, inland/metropolitan differences, and married/divorced or widowed variation. Spatial variations in mortality were not simply the result of occupational status differences as mortality from white collar groups was also elevated in inland areas. Spatial variation in mortality was not diluted due to the impact of immigration. CONCLUSIONS: Particular subgroups of people remained at risk and it is likely that differential access to ambulance services and travel time contribute, more particularly in inland rural areas. IMPLICATIONS: Groups at risk need to be more precisely targeted for preventative measures and public health initiatives, with particular implications for groups in inland rural areas, the Hunter Region, and outer lower income areas of Sydney. PMID- 10462856 TI - Trends and patterns of drug-caused mortality in Australia and Western Australia. AB - OBJECTIVE: To estimate the number of drug-caused deaths and compare patterns and trends of drug-caused mortality in Western Australia (WA) and Australia. METHOD: Aetiologic fractions were applied to mortality data for WA (1985-96) and Australia (1991-96) to estimate the number of deaths due to alcohol, tobacco and other drugs. RESULTS: Age-standardised rates (ASR) for smoking and alcohol-caused deaths fell significantly, while the ASR for deaths caused by other drugs increased significantly. About 85% of drug-caused deaths occurred among people aged 50 years and above. Tobacco-caused deaths mainly occurred among the elderly. Alcohol-caused death rates showed a minor peak in those aged 20-24 years then, from middle age, began to rise steadily, peaking in the oldest age group. However, deaths due to other drugs occurred mainly among young people, particularly those aged 25-34 years. These age differences were reflected in the person years of life lost (PYLL). People who died from other drugs lost, on average, about 31 years of life, twice as many as people who died from alcohol caused conditions and about six times the average number of PYLL per tobacco caused death. CONCLUSIONS: Nearly a fifth of all deaths were due to drugs and 80% of drug-caused deaths were due to tobacco smoking. The patterns and trends of drug-caused mortality in WA were similar to those Australia wide. IMPLICATIONS: Strategies highlighting the negative impact of tobacco and alcohol on health need to be maintained as well as programs which target illicit drug use. PMID- 10462857 TI - A single issue program in an isolated area: mammography screening in Darwin, NT. AB - OBJECTIVE: A process evaluation of the Northern Territory (NT) mammography program, NT Breast Screen (NTBS), during its initial 18 months of operation. METHODS: The study was undertaken in Darwin, NT, from December 1994 to May 1996. Clinical outcomes were obtained by reviewing computerised and manual program records to determine waiting times for results, recall rates and cancer detection rates. Client satisfaction was assessed by a questionnaire sent to all women with normal results over a 12-week period. General practitioner satisfaction was assessed by a questionnaire sent to all general practitioners in the region who had one or more clients who had attended the service. RESULTS: During this time, 2,882 screening mammograms were performed; 98 women were recalled for assessment (3.4%). Breast cancer was detected in 10 women (3.5 per 1000 women screened). The program was well accepted by clients and general practitioners. Performance criteria were not met for waiting times for results. CONCLUSIONS: NTBS faced challenges because of its small and dispersed population, a lack of local radiologists with mammographic experience and the conflict with other pressing health issues, particularly in Aboriginal health. Despite these challenges, the program functioned effectively during its initial 18 months. IMPLICATIONS: Mammography screening programs in isolated areas can function effectively. The constraints encountered by NTBS are likely to apply to similar programs. Issues identified requiring further research are the psychological consequences of long waiting times for results, and the prioritisation of mammography for Aboriginal women. PMID- 10462858 TI - Listeria awareness among new mothers in Western Australia. AB - OBJECTIVE: To assess awareness, knowledge and behaviour relating to Listeria among recent mothers (12 weeks post-partum) in Western Australia and determine the usefulness of a Listeria information pamphlet. METHOD: A postal survey of a random 10% sample of Western Australian mothers whose babies were born between 1 January and 13 April 1997. RESULTS: Of the 680 women surveyed, 509 (75%) responded and 451 (89%) respondents had heard of Listeria. The 11% who had not heard of Listeria had higher odds of living in a rural area, speaking a foreign language at home, having less formal education, being younger, having had an unplanned pregnancy, and having not taken folic acid supplements. Respondents who had seen the Listeria pamphlet had greater odds of correctly identifying foods at risk of Listeria. Of the women who had heard of Listeria, 90% had avoided certain foods during their pregnancy. Risk factors for not changing eating behaviour were similar to those for not having heard of Listeria. CONCLUSIONS: Factors associated with not having heard of Listeria, not having seen the pamphlet and not having changed behaviour were similar, suggesting that there may be a group of women who are less likely to be aware of Listeria or other health issues and/or may be resistant to health-related behaviour changes. IMPLICATIONS: The Listeria pamphlet is an effective medium for educating pregnant women about Listeria. Rural, young, single and non-English speaking background women may require a different or supplementary approach. PMID- 10462859 TI - Immunisation uptake, services required and government incentives for users of formal day care. AB - OBJECTIVES: To determine immunisation uptake in children attending formal day care prior to the introduction of certificates (state) and parent incentives (federal), and to document parent and child carers' attitudes to these strategies. METHOD: In 1997, 60 child care centres and 300 family day carers in suburban Melbourne were randomly sampled. Immunisation dates, service use and preference, and views on government incentives were obtained from parents of children under three years of age. RESULTS: From 2,454 eligible children, information was obtained for 1,779, of whom 84% (95% CI 82-86) were completely immunised. Low income (OR 1.8, 95% CI 1.2-1.9, p < or = 0.001) and larger family size (OR 1.8, 95% CI 1.2-2.7, p = 0.002) and only ever using a doctor (OR 1.6, 95% CI 1.1-2.3) was associated with incomplete immunisation. Main reasons for delaying immunisation were occurrence of minor illness and work commitments. Families would prefer immunisation services at Maternal and Child Health visits (39%), evening sessions (22%) and at day care (22%). Immunisation uptake could increase to 94% if those receiving Childcare Assistance (67%) immunised their children on time but would increase to 87% if these incentive only motivated those for whom Childcare Assistance was essential (15%). While 98% of day care co ordinators and 71% of family day care co-ordinators documented immunisation status at commencement of child care, only 51% and 33% respectively regularly updated this information. CONCLUSION: Providing client-focused, flexible immunisation services and government incentives and legislation may work together to boost immunisation levels for those in formal child care. PMID- 10462860 TI - Seroprevalence of markers for hepatitis B, C and G in male and female prisoners- NSW, 1996. AB - OBJECTIVES: 1. Establish the prevalence of markers for hepatitis B (HBV), C (HCV) and G (HGV) in a sample of male and female inmates. 2. Examine exposure to multiple viruses. 3. Compare risk factors for HGV infection with known risk factors for HBV and HCV. DESIGN: Cross-sectional random sample stratified by sex, age and Aboriginality. Inmates were screened for three hepatitis markers. Participants were 789 inmates (657 male, 132 female) in 27 correctional centres in New South Wales, 1996. RESULTS: Overall detection of each of the three screening markers was 35% for HBV, 39% for HCV and 10% for HGV. Exposure rates were higher in female prisoners than males. Increased rates of anti-HBc were observed in Aboriginal inmates compared with non-Aboriginals (54% cf. 27%); anti HCV and HGV-RNA were comparable between the two groups (36% cf. 41% and 9% cf. 10%). Markers were significantly higher in female injecting drug users (IDU), particularly HCV (90% cf. 66%). Thirty-five per cent of inmates were unaware of their HCV status. For HBV, 72% did not self-report past or present exposure despite serological evidence to the contrary. The multivariate analysis identified Aboriginality, long-term injecting and injecting while in prison as risk factors for HBV. HCV risk factors were female sex, non-Aboriginality, institutionalisation and IDU-associated behaviours. For HGV, female sex and previous imprisonment were significant risk factors but IDU was not. CONCLUSIONS: Blood-borne hepatitis viruses are common in prison inmates, particularly females (HBV, HCV and HGV), Aboriginals (HBV) and IDU (HBV and HCV). Infection can be related to a number of risk factors, which appear similar for HBV and HCV, but distinct from HGV. PMID- 10462861 TI - Cross-cultural research: trying to do it better. 1. Issues in study design. AB - OBJECTIVE: To discuss a range of strategies to address the methodological and practical challenges in designing cross-cultural public health studies. METHOD: The Mothers in a New Country (MINC) Study was an interview study of 318 Vietnamese, Turkish and Filipino recent mothers exploring their views of maternity care and the early months of motherhood. It was carried out in Melbourne between 1994 and 1997. Sampling, recruitment, retention and representativeness all pose problems for studies involving non-English-speaking background immigrant populations, as do selection, training and support processes for bicultural interviewers. These issues are discussed with reference to the strategies undertaken to tackle them in the MINC Study. RESULTS: In the MINC Study, a systematic approach to sampling and recruitment, combined with a flexible and sensitive study protocol were largely successful both in achieving in adequate sample size and a largely representative study sample. Similarly, paying significant attention to the selection, training and ongoing support of the biocultural interviewers employed on the study contributed greatly to its successful completion and enhanced confidence in the study findings. IMPLICATIONS: Both researchers and funders need to take seriously the implications of the many methodological and practical issues involved in designing sound cross-cultural public health studies. In particular, there are major implications for study costs and timelines. However, the benefits to be gained are significant. PMID- 10462862 TI - Cross-cultural research: trying to do it better. 2. Enhancing data quality. AB - OBJECTIVE: To describe the dilemmas for cross-cultural research in translating study instruments and implementing quality assurance methods, drawing on strategies utilised in the Mothers in a New Country (MINC) Study. METHOD: To translation of study instruments in the MINC Study included: forward and back translations, a bilingual group review process, consultation with bilingual content experts, piloting of different versions of translations, a process for exploring unresolved difficulties and caution in interpreting unusual study findings. Interview quality was assessed by: 1. An ongoing review of interviewer prepared English coding schedules to ensure completeness of data and identify problems with interview administration. 2. Analysis of fully translated transcripts of six randomly selected early interviews to assess the accuracy and consistency with which questions were asked. 3. A comparison of data sources for 45 randomly selected interviews (original language interview schedules, English coding schedules and translated interview transcripts) to determine the rate and nature of discrepancies. RESULTS: Translation strategies that went beyond simple forward and back translations achieved more reliable and appropriate translations. The complexity of language and cultural differences sometimes still meant less than satisfactory results. Interview tapes played an important quality assurance role, enabling feedback to the interviewers and providing a basis of comparison for identification of data discrepancies. IMPLICATIONS: Ensuring good data quality in cross-cultural research is both critically important and difficult. Open discussion of the problems and concerted efforts to deal with them would benefit future research. PMID- 10462863 TI - Knowledge and perceptions about breast cancer incidence, fatality and risk among Australian women. AB - OBJECTIVE: Breast cancer is the most common fatal cancer among Australian women. This study aimed to provide an accurate national picture of women's understanding of breast cancer incidence, fatality and risk. METHOD: A telephone survey explored the knowledge and perceptions of a sample of 2,935 Australian women in relation to breast cancer incidence, fatality, risk factors, risk perception and level of concern. RESULTS: Australian women were well aware of breast cancer in general terms, however, there were major aspects of incidence and risk which were poorly understood. Only 5% of women nominated age as a risk factor, and only one third were able to make an approximately correct estimate of the incidence of breast cancer in Australia. CONCLUSIONS AND IMPLICATIONS: Recommendations for future information campaigns include targeting understanding of lifetime risk of developing breast cancer, age as a risk factor, survival from breast cancer and the need to separately address the perceptions of older versus younger women. PMID- 10462864 TI - SF-36 health survey reliability, validity and norms for New Zealand. AB - OBJECTIVE: To assess the acceptability, reliability and validity of the SF-36 health survey in the New Zealand population and provide key population norms. METHOD: The SF-36 questionnaire was part of the 1996/97 New Zealand health survey, a cross-sectional, nationally representative survey of 7,862 adults (15 years and over). RESULTS: Overall, in the New Zealand population the questionnaire performed as well as or better than in other national surveys, but there was variability in data completeness across subgroups, and responses were skewed towards the healthy end of the scales. Males scored higher than females on almost all scales; increasing age was associated with decreasing scores (with the exception of the mental health scale); and New Zealand Europeans tended to report better health than the other ethnic groups. CONCLUSIONS: Satisfactory psychometric performance was demonstrated for the SF-36 in the New Zealand population, but researchers need to find ways of increasing data completeness in population groups shown here to have lower completion rates. The questionnaire may be better at discriminating patient rather than population subgroups. The SF 36 normative data confirm in kind, if not in degree, population subgroup disparities in health status observed using objective measures. IMPLICATIONS: Overall, the SF-36 questionnaire appears to be a valid and reliable measure of health-related quality of life for the New Zealand population. However, this paper highlights issues for researchers using the SF-36, such as the skewed nature of responses obstained in a population sample. PMID- 10462865 TI - Are smoke-free policies implemented and adhered to at sporting venues? AB - OBJECTIVE: This study investigated the level of implementation of and adherence to smoke-free policies in two major sporting venues in Perth. METHOD: Smoking status and attitude toward the smoke-free policies in the venues were determined in a random sample of spectators as they entered each venue. An observational study of randomly selected non-smoking seated areas in each venue was conducted on the same day to determine compliance with smoke-free policies. A butt-count was conducted to validate these observations. RESULTS: There was a high level of both awareness and agreement with the smoke-free policies, however, this level of agreement was higher in non-smokers than smokers. The results of the observational study and the butt-count indicated that the policies were implemented and there was a high level of adherence with the smoke-free policy at both venues. CONCLUSION: The results provide further evidence that smoke-free policies in sporting venues are both supported and adhered to by spectators. IMPLICATIONS: The implementation of the smoke-free policies at venues is highly effective in protecting non-smokers from the effects of environmental tobacco smoke. PMID- 10462866 TI - Specificity of notification for tuberculosis among screened refugees in NSW. AB - OBJECTIVE: Epidemiological surveillance of tuberculosis (TB) in Australia is dependent on the accuracy of the notification data. We have investigated the specificity of TB notification for the diagnosis of this disease. METHOD: We used data from notifications to the NSW Department of Health to identify members of a cohort of refugees who were screened between 1984 and 1993 and subsequently developed TB during that period. We reviewed notification data and, in most instances, case notes and x-ray films, to independently confirm or refute the diagnosis of TB. RESULTS: Two hundred and fifty members of the cohort were identified in the notification database. After excluding refugees on treatment prior to arrival in Australia, and those who were notified as 'quiescent' and 'atypical' cases, there were 189 cases notified as active TB. There was evidence to support the diagnosis of active TB in 125 cases (66%) and evidence that subjects did not have active disease in 60 cases (32%). We could not determine the status of the remaining four notified cases. CONCLUSION: This study has shown that, in a population of refugees subject to screening, nearly one-third of cases notified as active tuberculosis from the study population were actually not active tuberculosis cases. IMPLICATIONS: The use of the TB notification database may result in overestimation of the incidence of TB in population groups who are subject to active screening. PMID- 10462867 TI - The timeliness of notification of clinically suspected cases of dengue imported into north Queensland. AB - OBJECTIVE: Evaluate the timeliness of notifications by medical practitioners of clinically suspected dengue importations into north Queensland (NQ). Describe the features the disease and determine the likely duration of viraemia prior to implementating public health measures. METHOD: Since December 1994, the Tropical Public Health Unit (TPHU) has maintained a register of all imported cases of dengue into NQ. Only confirmed viraemic cases were included in the study. Timeliness of notifications was defined as the interval between the first medical consultation in NQ and notification to TPHU. An Epi info database was developed and used to analyse data. RESULTS: There were 31 confirmed dengue importations into NQ during the study period, including all four dengue serotypes. The largest source (39%) was Papua New Guinea. The median time for notifications was 5.5 days. Doctor notifications ranged from 0 to 21 days (median 2 days), remaining notifications range from 1 to 42 days (median 10 days) (p < 0.05). The mean duration of viraemia of public health importance was 7 days (2-12 days). CONCLUSIONS: Of concern, doctors failed to notify > 50% of suspected cases and only 26% of notifications were received within 48 hours. Notification delays led to prolonged viraemia of public health importance increasing the potential risk of secondary infections. IMPLICATIONS: General practitioners need to promptly notify all clinically suspected cases of dengue in travellers recently arrived in NQ. The TPHU intends to reemphasise to general practitioner the importance of timely notifications and develop an orientation package for new doctors. PMID- 10462868 TI - One year follow-up of an emergency department protocol for abused women. AB - OBJECTIVE: To evaluate if the positive effects of an initially successful emergency department protocol of care for victims of partner abuse (PA) were maintained at one year follow-up. METHOD: A community intervention trial compared two public hospital EDs in Auckland, NZ, at baseline (BL), following implementation of a PA intervention (PI), and at one year follow-up (FU). The initial intervention involved training staff at one ED in a protocol for the identification and acute management of abused women. Later, an on-site protocol co-ordinator worked to ensure that new staff received training in protocol use. Outcomes were assessed by reviewing a random sample of women's medical records. Identification of PA was assessed for each record on a yes/no basis. Identified cases were classified as 'confirmed' or 'suspected' PA. Acute management was assessed by ascertaining staff use of interventions and documentation of abuse. RESULTS: Approximately equal numbers of records were reviewed at each ED at BL, PI and FU (total n = 10,961). Significant interactions were found for classification of PA and acute management offered to victims between EDs over the different study phases. However, the increase in identification of confirmed cases and the improved acute management offered to identified victims found at the intervention ED in the PI phase were not maintained at follow-up. CONCLUSION: Although staff training in the use of protocols for victims of partner abuse can result in improved care, these effects were not maintained. IMPLICATIONS: More intensive institutional efforts are needed if protocol benefits are to be maintained. PMID- 10462869 TI - A nursing home outbreak of Clostridium perfringens associated with pureed food. AB - OBJECTIVE: To investigate and ascertain the source of a nursing home outbreak of gastroenteritis in Melbourne in 1997. METHOD: A retrospective cohort study. We obtained fecal and food samples and environmental swabs, and reviewed food handling. RESULTS: There were 25 cases, and 21 of these had their meals pureed (liquidised). The relative risk for eating pureed food and becoming ill was 5.8 (95% CI 2.2-15.4). Clostridium perfringens and its enterotoxin was detected in nine fecal samples. Samples of pureed food tested positive for coliforms. CONCLUSIONS: This outbreak was caused by C. perfringens contaminating pureed food. Food liquidising provides opportunities for re-contamination of cooked food through the use of contaminated equipment and deficiencies in food handling practices. Pureed foods should be reheated to 70 degrees C after liquidising to inactivate pathogens. IMPLICATIONS: Health care facilities need to take specific precautions to manage the special hazards involved in preparing pureed food. PMID- 10462870 TI - Social capital and its relevance to health and family policy. AB - It is important to consider the global context in which we operate and in which the association between society and health is to be explored. We are in a democratic world, with an economy that emphasises the market and a post-modern culture. The ancient elements of state, market and community are discernible in avaricious combinations and, if we are seeking a just social order, it is important that we apply appropriate analytical methods to social understanding. While social capital is an arresting term, its ambiguity limits its broad applicability and even makes it dangerous. Sir Richard Doll and his colleague Richard Peto once described epidemiological insight as a boundary-setting exercise, delimiting territory within which basic and clinical science can explore mechanisms. When it comes to matters of social and personal well-being, the same approach may serve us well. Social capital, defined differently by everyone who uses it, must be given some stability and be subject to good quality epidemiological research, not too dissimilar to that which has underpinned epidemiology's immense success in public health over the decades. Despite social capital's complexity, there are growing efforts to measure it and relate it to desirable social functions, but the ability of social capital to capture fully the subtle interplay of individuals and society so essential for their health and happiness is questionable. PMID- 10462871 TI - Social capital and community development: practitioner emptor. AB - Social capital has become the latest 'flavour of the month'. There is considerable disagreement over what the term means, and calls for theorising and measurement of the construct. Health promoters, among others, are being challenged to re-construct their efforts around this still contested idea. Social capital doesn't exist, but is being created by those aspects of social relations particular theorists or researchers choose to study in its name. The choice of these relations is directed by ideology. To those aligned more with neoliberal, market-driven ideology, social capital is a means to the end of economic growth, something that can pick up the slack of privatised or reduced public services. To those aligned more with social justice and communitarian ideology, social capital is an end in itself, requiring the defence of strong, egalitarian state intervention into market practices that create inequalities. Community development is one of many state interventions used to buffer market-generated inequalities. Social capital may be a useful concept for practitioners, researchers and policy makers in bring the missing 'social' into economic and fiscal policy debates. But its use should be approached cautiously as a construct of potential strategic value. It should not confuse all of the previous 'good' work undertaken in the name of empowerment and community capacity. PMID- 10462872 TI - Drinking and diving: public health concerns. AB - Stage diving and crowd surfing are an established part of clubs and open-air concerts. Despite policy attempts to reduce accident prevalence rates, teenagers and young adults are at risk in these often volatile environments. Attempts to promote harm minimisation are discussed. PMID- 10462873 TI - Evidence-based public health practice. PMID- 10462874 TI - Lipoproteins, atherogenicity, age and risk of myocardial infarction. PMID- 10462875 TI - Olympics visitors need to be told about the dangers of the Australian surf. PMID- 10462876 TI - Measles encephalitis in Victoria, 1962-96: down but not out. PMID- 10462877 TI - The Canadian Model of Occupational Performance applied to females with osteoporosis. AB - Osteoporosis is a prevalent disease among older women. It results not only in physical limitations, but may raise spiritual and affective issues that can interfere with a person's ability to perform routine occupations. In addition, the social, cultural, physical, and institutional elements of the environment play a vital role in influencing the choices individuals make and how they manage symptomatic osteoporosis. The purpose of this paper is to examine the occupational impact of osteoporosis on post-menopausal women by applying the Canadian Model of Occupational performance (Canadian Association of Occupational Therapists, 1997) as an organizational framework to existing knowledge of this disease. PMID- 10462878 TI - Enabling occupational performance: optimal experiences in therapy. AB - Occupational therapists believe that engagement in occupation contributes to health through an individually balanced use of time, a positive focus for one's physical and mental energy, and the provision of a sense of purpose. Flow is a construct which describes optimal experiences or enjoyment in everyday activities. A review of the literature suggests that the theory of optimal experience is complementary to occupational therapy beliefs and that an understanding of the flow experience may contribute to our understanding of human occupation. Specifically, flow may be useful in understanding those aspects of the occupation, environment and person that contribute to a "just right" challenge, and to enabling occupational performance through enjoyable, structured and purposeful activity. Occupational therapists are encouraged to explore whether optimal experiences facilitate occupational performance for individuals with a disability. Future research could explore whether the occupational opportunities available to persons with a disability provide the degree of challenge required to elicit the optimal experience. Finally, research could explore whether the client-driven selection of meaningful occupation, and therapist enablement of the "just right" challenge, influences optimal experience, occupational performance, and life satisfaction for those with a disability. PMID- 10462879 TI - [Questionnaire on the satisfaction of persons with lower-limb amputations towards their prosthesis: development and validation]. AB - The satisfaction of persons with lower-limb amputations towards their prosthesis constitutes a critical factor in the use of the prosthesis. In order to evaluate a person's satisfaction, the SAT-PRO, a self-administrated questionnaire was developed. The questionnaire includes 15 items developed on the basis of the most significant criteria used by the person when selecting a technical aid. These criteria were measured using an ordinal categorical four-level scale. The validation of the SAT-PRO was established from a sample consisting of 61 people with below-knee or above-knee amputations, aging from 60 years and older. The internal consistency of the instrument is high (Cronback's alpha coefficient is 0.90) and the test-retest reliability coefficient (0.97) indicates a very good consistency of the questionnaire over time. Simple and multiple correlations were used to evaluate construct validity. The degree of use of the prosthesis and the feelings of depression, amongst the measured variables, are the best indicators of the satisfaction of the amputees toward their prosthesis. PMID- 10462880 TI - Reliability and validity of the Toglia Category Assessment Test. AB - The aim of this study was to establish interrater reliability, internal consistency reliability, and both concurrent and discriminant validity for the Toglia Category Assessment (TCA), a test of cognitive categorization. This study was based on an examination of categorization skills in clinical samples, using both the Toglia Category Assessment as well as the Riska Object Classification (ROC) test. The study sample consisted of 35 adult persons with brain-injury and 35 persons with chronic schizophrenia (n = 70). Correlations between scores of the two different tests and the two different samples were computed using Pearson's Product Moment Correlation Coefficients. Both inter-rater reliability, internal consistency, and concurrent validity were established for the TCA. These results suggest that the Toglia Category Assessment and Riska Object Classification tests may address different cognitive abilities. The Toglia Category Assessment appears to be a more sensitive test than the Riska Object Classification in that it provides an elaborate cognitive profile. PMID- 10462881 TI - Frames of reference utilized in the rehabilitation of individuals with eating disorders. AB - An occupational therapist's role with clients diagnosed with eating disorders, both anorexia nervosa and bulimia nervosa, has been described throughout the literature. However, the frames of reference and treatment approaches that occupational therapists implement have not been clearly established or validated. This paper outlines the symptomatology of anorexia nervosa and bulimia nervosa, and critically reviews the current literature concerning the frames of reference and treatment approaches used by occupational therapists when intervening with this population. The literature reviewed indicates that therapists are using a variety of frames of reference and treatment approaches. There appears to be an emphasis on the psychoanalytical and cognitive-behavioural frames of reference, although there is a lack of empirical evidence in regards to all frames of reference and treatment approaches. Reasons for the lack of current research with this population, and possible future areas of research are suggested. PMID- 10462882 TI - Assessment and intervention with clients with apraxia: contributions from the literature. AB - Individuals with praxis problems encounter difficulties engaging in occupation, and the occupational therapists who work with these clients are challenged to provide intervention that enables occupational performance. This extensive review of the current literature provides clinicians with information regarding the relevant descriptions and suggested mechanisms of apraxia. Errors noted in performance, different classification systems and the implications of the current knowledge will be discussed. The paper concludes by providing information for clinicians regarding occupational therapy evaluation and intervention strategies for praxis problems. PMID- 10462883 TI - The new professional--the nexus of healthcare trends. PMID- 10462884 TI - Defining and developing professionalism. AB - During the development of a new occupational therapy curriculum, professionalism was identified as a core process component essential to occupational therapy practice. A group, comprised of faculty, clinicians, and students, was charged to examine professionalism and to make recommendations for curriculum planning and development. However, a consistent description or definition of professionalism was lacking in the literature. Defining professionalism was, therefore, the first task of the group. A schematic representation of professionalism was developed through a review of the literature and qualitative analysis of information obtained from discussion groups focussed on professionalism. In this paper, the schematic representation of professionalism will be presented as well as information about two supported self-study courses entitled, Fostering Professional Development and Becoming a Professional. A professional portfolio guide for the student occupational therapist will also be described. Future implications and directions for fostering professionalism will be discussed. PMID- 10462885 TI - Application of the Person-Environment-Occupation Model: a practical tool. AB - Occupational therapy focuses on complex dynamic relationships between people, occupations and environments. Therapists must clearly communicate their practices and how their practice influences outcomes. This paper explores applications of the Person-Environment-Occupation Model (Law et al., 1996) in occupational therapy practice, and delineates how this particular model helps therapists to conceptualize, plan, communicate and evaluate occupational performance interventions. Three case studies illustrate how the model can be used by occupational therapists to systematically approach analysis of occupational performance issues while considering the complexities of human functioning and experience. The ways in which the model facilitates communication within and outside occupational therapy are explained. The Person-Environment-Occupation Model is offered as a tool for therapists to use in client(s)-therapist alliances to enable clients to successfully engage in meaningful occupations in chosen environments. PMID- 10462886 TI - [Test-retest and inter-rater reliability of the French version of the Ontario Society of Occupational Therapy (OSOT) Perceptual Evaluation]. AB - This article presents the results of a study conducted to verify the test-retest and inter-rater reliability of the French version of the Ontario Society of Occupational Therapy (OSOT) Perceptual Evaluation. Designed to evaluate the perceptual deficits in patients with brain injuries, this tool uses a 5-points scale (0-4) to measure 18 different tasks. The scores obtained for each task are added to establish a total score. In the early 90s, the instruction manual of the OSOT Perceptual Evaluation was translated in French by a group of occupational therapists from l'Institut universitaire en geriatrie de Sherbrooke. To ensure the reliability of this version, a study was conducted to determine the test retest reliability and the simultaneous inter-rater reliability. Thirty-two francophone subjects with brain injuries were each evaluated twice by the same therapist to determine the test-retest reliability of this tool. At one of the two encounters, a second therapist completed the score sheet to verify the simultaneous inter-rater reliability. The results show that, despite a few weak kappas' scores for certain tasks, the test-retest reliability and the inter-rater reliability of the total score were excellent (test-retest reliability: intra class correlation coefficient = 0.93, with a confidence interval of 0.87 to 0.97; and inter-observer reliability: = 0.98, with a confidence interval of 0.97 to 0.99). The findings of this study show that the French version of the OSOT Perceptual Evaluation can therefore be used confidently by francophone occupational therapists. PMID- 10462887 TI - A comparison of interface pressure readings to wheelchair cushions and positioning: a pilot study. AB - Occupational therapists are often involved in assessing the seating needs of clients who have quadriplegia. One component of this process involves determining the appropriate use of wheelchair cushions and pressure relieving techniques in the prevention of pressure sores. The purpose of this study was to assess the effect of tilt and recline positioning and various wheelchair cushions on interface pressures at the ischial tuberosities and the sacrum of two subjects with C5 quadriplegia. In addition, interface pressures under bony prominences were assessed in the personal driving position (when the subject enters), neutral, 35 degree tilt, 45 degree tilt, and maximum recline (150 degrees). The results of this study indicate that for the two subjects evaluated, higher average interface pressures at the ischial tuberosities were obtained when using one of the three cushions under scrutiny. Furthermore, the general trend observed with both subjects is a reduction of pressure readings at the ischial tuberosities with tilt and recline positioning (especially with 45 degree tilt and 150 degree recline). The findings of this study are in keeping with previous studies which identified that individual and ongoing assessment is essential in providing the best cushion and pressure relief techniques for individuals. PMID- 10462888 TI - Bovine spongiform encephalopathy and new variant Creutzfeldt-Jakob disease: an overview. AB - About 10 years after bovine spongiform encephalopathy (BSE) appeared in British cattle, a new variant of Creutzfeldt-Jakob disease (nv-CJD) was described in the United Kingdom. This new disease is distinguishable from classical CJD in its aetiology, epidemiology, clinical profile, and neuropathology. The emergence of nv-CJD raised fears of a causal relationship between BSE and nv-CJD and of a human epidemic of indeterminate size. This paper reviews our knowledge of this group of diseases, and examines recent scientific evidence, which indicates that BSE is indeed the source of nv-CJD, and discusses the public health implications. PMID- 10462889 TI - The application of near patient testing to microbiology. AB - Near patient testing (NPT) was the norm in days when urine was examined by smell and taste. More recently, general practitioners and physicians in genitourinary medicine began to use light microscopes in their consulting rooms to examine urine for pus cells and urethral and other swabs for pathogens. Increasing knowledge has led to specialisation, however, with clinicians obtaining specimens for examination by others. Improved technology has speeded up the practice of medicine, raising expectations of patients and doctors alike, and reductions in the size and expense of testing instruments have made a renaissance of NPT possible. Such a rebirth has already been seen in high dependency units and neonatal intensive care units, where arterial blood gases and serum bilirubin have to be tested in less time than it would take a sprinter to reach the laboratory. People with diabetes, rushing about in the community, stop and test their own blood glucose to determine the ideal dose of insulin, and patients with asthma measure peak expiratory flow rates to titrate doses of inhaled and oral corticosteroids. To what extent has NPT developed in microbiology? General practitioners have nitrite dipsticks and dipslides with which to identify urinary tract infections and elsewhere in this issue the prospect for testing for Helicobacter pylori infection is discussed. Do-it-yourself HIV testing kits can be bought in some countries. Are these desirable developments for communicable diseases, the results of whose investigation are used not only to benefit the individuals tested but also to monitor trends in populations and determine policies for the prevention and control of infection? If NPT is desirable, or inevitable, in microbiology, how can it be developed so as to ensure a high quality service both for patients and the population? This review considers the implications of NPT in the field of communicable diseases for microbiology laboratories, quality assurance, accreditation, and the legal framework in which medical devices are used. PMID- 10462890 TI - Escherichia coli O157 incident associated with a farm open to members of the public. AB - Three children, one who lived on an open farm and two who visited the farm in school parties, developed Vero cytotoxin producing Escherichia coli (VTEC) O157 infection in 1997. All had been in direct contact with animals. All were admitted to hospital with diarrhoea and two developed the haemolytic uraemic syndrome, one of whom developed severe neurological impairment. E.coli O157 phage type 21 that produced Vero cytotoxin 2 were isolated from the three children and from a goat paddock and two cows at the farm. The isolates were indistinguishable by molecular typing. The farm closed voluntarily for six weeks while recommendations to reduce the exposure of visitors to faecal contamination and to improve hygiene procedures (especially handwashing) were implemented. More research is needed into all aspects of VTEC O157 on farms, including the natural history of carriage in animals, the organism prevalence and factors that affect infectivity to humans. Further consideration is needed about the role of open farms in zoonoses, and how open farms can be assisted in reducing risks for visitors. The public need to be educated about the risks and about their responsibilities, which include the need to wash hands thoroughly and to avoid hand to mouth contact when visiting farms. PMID- 10462891 TI - Outbreak of gastroenteritis associated with contamination of a private borehole water supply. AB - An outbreak of gastroenteritis affected 58 of 700 people served by a private water supply at a biological research institute located in a village. No cases were detected in 250 residents of the same village served by a public water supply over the same period. Consumer complaints of tainting and laboratory evidence of bacterial and chemical contamination were obtained for the private water supply, but not for the public water supply. The outbreak was probably caused by contamination from a nearby sewer of a borehole used for the private supply. The outbreak showed how a large, private water supply posed a substantial risk to public health. The regulatory framework for such water supplies should be modified to ensure their safer design and operation. PMID- 10462892 TI - Common source outbreak of salmonellosis in a food factory. AB - Seventy-three employees at a food processing factory employing 2700 staff reported vomiting, diarrhoea, or abdominal pain between 30 July and 3 August 1997. Salmonella enteritidis phage type (PT) 4 was isolated from 47 symptomatic cases and five asymptomatic canteen staff. The epidemic curve suggested a point exposure to a common source: 60% of cases reported illness on 31 July. An uncooked dessert containing raw shell eggs was identified as a possible vehicle of infection. Caterers are reminded that pasteurised egg should be substituted for shell egg in dishes that are eaten raw or only lightly cooked. PMID- 10462893 TI - Investigation of an outbreak of gastroenteritis at a hospital for patients with learning difficulties. AB - Eighty of the 460 patients and staff (attack rate 22%) in a long-stay hospital for patients with learning difficulties became ill in a general outbreak of gastrointestinal infection that followed a buffet style party on one ward. Illness in the cohort of 47 resident patients, relations, and staff who attended that party was associated with having eaten ham, coleslaw, bread rolls, and cheese and pineapple on sticks. Food from the hospital kitchen was supplemented by food brought in and prepared on the ward by staff and patients. The investigation suggested that food items were contaminated either during preparation by staff and patients or during the party when people served themselves. Microbiological and virological investigations were negative, but small round structured virus was thought to be responsible. Hospitals could do more to prevent outbreaks of foodborne infection by undertaking risk assessment as required by the Food Safety Act (General Food Hygiene) Regulations 1995. Purchasers of hospital infection control should ensure compliance with this legislation. PMID- 10462894 TI - An outbreak of campylobacter food poisoning at a university campus. AB - Twelve cases of Campylobacter jejuni infection were identified on a university campus in the first 12 days of November 1997. Consumption of food from a single outlet, where poor food handling practices were identified, was statistically associated with infection. The epidemiological evidence suggested that what presented as a point source outbreak was a series of small cross contamination incidents associated with multiple strain types. The nature of this outbreak suggests that current surveillance systems are inadequate for the identification of campylobacter outbreaks. Enhanced surveillance should be introduced to identify case clusters of campylobacter infections and public health professionals should remember the risks of cross contamination associated with ubiquitously contaminated foodstuffs, especially raw meats and poultry. The role of strain typing in campylobacter epidemiology is as yet undefined. PMID- 10462895 TI - Outbreak of hepatitis A in Rotterdam associated with visits to 'darkrooms' in gay bars. AB - An unexpectedly large number of hepatitis A virus (HAV) infections were notified among homosexual men in Rotterdam in the first five months of 1998. A case control study was conducted to investigate the hypothesis that this outbreak was associated with sexual practices and to collect information with which to focus preventive activities. Notified cases and controls selected from male members of a gay sports club completed anonymous questionnaires about known risk factors for HAV infection and sexual behaviour. Single variable analysis showed that HAV infection was associated with sexual contact with anonymous sex partners (odds ratio (OR) 4.6; 95% confidence interval (CI) 1.0-23.2) and with visits to 'darkrooms' in gay bars (OR 6.2; 95% CI 1.5-26.8). A negative association with travel abroad to western countries was observed (OR 0.4; 95% CI 0.1-1.3). The associations with visits to darkrooms (OR = 9.2; 95% CI 1.6-52.4) and travel abroad to western countries (OR 0.1; 95% CI 0.02-0.9) remained significant in a multivariable logistic regression analysis. This risk group was targeted for health education and vaccination and darkroom owners were advised to provide hygiene facilities. PMID- 10462896 TI - A district survey of vaccine cold chain protection in general practitioners' surgeries. AB - Failure to ensure that vaccines are kept within a prescribed temperature range at all times can reduce their potency and cause primary vaccine failure. A postal survey of 103 general practices in a health district to assess vaccine handling and storage yielded 75 responses (73%). Poor practice was identified in receipt and storage of vaccines, temperature monitoring and control, management of vaccines during immunisation sessions, and disposal of partly used vaccines. The data suggest that the vaccine cold chain is not maintained with the degree of care necessary for safe practice. National guidelines need to be implemented conscientiously by all those involved with immunisation programmes if the effectiveness of vaccines is to be guaranteed. PMID- 10462897 TI - Measles, mumps, and rubella vaccine coverage in 2 year old children in East Lancashire--better than it looks. AB - A study population of 1850 children resident in East Lancashire born between 2 July and 1 October 1994 was obtained from local child health information systems (CHIS) and family health service (FHS) general practitioner registration data in March 1997 to determine the accuracy of reported measles, mumps, and rubella (MMR) vaccine coverage in 2 year old children registered with East Lancashire general practitioners. The reported MMR immunisation coverage was 89.7%, but the observed coverage was higher at 95.5% (95% confidence interval (CI) 93.9-97.3%). Small practices (3 Pounds GPs) achieved significantly higher MMR coverage (97.1%; 95% CI 95.8-98.0%) than large practices (93.5%; 95% CI 91.5-95.1%). Significantly higher MMR coverage was found in practices that used CHIS recall systems (96.2%; 95% CI 95.1-97.1%) than those using their own recall methods (91.2%; 95% CI 87.0 94.4%). Logistic regression showed that MMR vaccine coverage was independently predicted by practice size (odds ratio (OR) 2.5; 95% CI 1.5-4.0), recall method (OR 2.3; 95% CI 1.4-3.8), and relative deprivation (OR 1.7; 95% CI 1.0-2.6). Actual MMR coverage in 2 year old children in East Lancashire was significantly higher than reported, mainly because of inaccuracies in the CHIS database. Methods to improve the transfer of data on immunisation from practices to population databases should be explored. PMID- 10462898 TI - A survey of the implementation of health service guidelines on arrangements for infection control in health trusts in the West Midlands. AB - A good hospital infection control programme can reduce hospital acquired infection, which causes considerable morbidity, mortality, and cost. NHS trusts in the West Midlands were surveyed to assess progress in implementing national infection control guidelines. All 25 acute trusts replied but only 13 of 21 trusts for the community or for mental health (MH). Twenty-four acute trusts had access to an infection control nurse (ICN), but ICNs were responsible for an average of 520 acute beds, twice as many as usually recommended. Seven (of 13) community/MH trusts had only informal arrangements for access to an ICN. Six acute trusts had no formal arrangements for covering ICN leave. All 25 acute trusts had access to an infection control doctor, but on-call cover in eight required this individual to be permanently available. All acute trusts had a hospital control of infection committee, but in only 10 did a senior member of management regularly attend. Twenty-four acute trusts had an outbreak control plan but only 13 had been updated as recommended. The NHS performance management structure needs to be utilised to ensure that these deficiencies are rectified. It would be wise to investigate implementation of national guidance in NHS trusts in other regions. PMID- 10462899 TI - A comparison of six commercial kits for Helicobacter pylori detection. AB - The British Society of Gastroenterologists suggests that dyspeptic patients under 45 years of age should be screened serologically for Helicobacter pylori infection, to reduce endoscopy workload. We have compared the sensitivity, specificity, and predictive value of six commercial serological kits intended for pre-endoscopy screening for H. pylori with histopathology and culture in 82 dyspeptic patients, 35 of whom were H. pylori positive. The kits' sensitivities were as follows: Bio-Rad GAP 100%, Helico-G 100%, Premier 97%, and Pyloriset EIA G 94%. Poor specificity of the ELISA kits--Bio-Rad GAP 67%, Helico-G 67%, Premier 85%, and Pyloriset EIA-G 76%--was due to previous treated or cleared H. pylori infection. Allowing for previously documented H. pylori infection or peptic ulcer improved specificity--Bio-Rad GAP 84%, Helico-G 84%, Premier 100%, and Pyloriset EIA-G 90%. The Pyloriset Dry latex kit had a higher specificity (86%) but a lower sensitivity (75%) than the Oxoid latex kit (specificity 70%, sensitivity 94%). The qualitative Premier Launch kit had the best overall results (and was the easiest ELISA to perform). Reliable serological diagnosis of H. pylori is now suitable for screening dyspeptic patients. PMID- 10462900 TI - Surveillance of neonatal group B streptococcal infection in Sunderland. AB - Surveillance of neonatal group B streptococcus infection in Sunderland identified 15 confirmed cases (with positive cultures from blood or CSF) in three years from 1995 to 1997, equivalent to an incidence of 1.42 per 1000 live births, not much lower than the estimate of 1.8 used in the United States to justify the introduction of preventative policies. Confirmed early-onset cases may represent only a fraction of the true number of cases, and a modified risk factor-based policy was introduced in Sunderland in June 1998. PMID- 10462901 TI - Undercooked hens eggs remain a risk factor for sporadic Salmonella enteritidis infection. AB - The prevalence of salmonella contamination in hens' eggs remains high. A case control study of sporadic cases of Salmonella enteritidis infection in Wales in 1997 confirmed the association of infection with consumption of raw or lightly cooked eggs. PMID- 10462902 TI - Microbiological quality of drinking water from office water dispensers. AB - A survey of the microbiological quality of water sampled from office dispensers in Merseyside found half of the samples to be either unsatisfactory (23/56) or unacceptable (5/56) in terms of guidelines from the Automatic Vending Association of Britain. Factors that appeared to be linked to unsatisfactory microbiological quality were time between filling and sampling and filling on site. PMID- 10462903 TI - Domiciliary vaccination: a practice overlooked. PMID- 10462904 TI - Automation of the polymerase chain reaction. Part I. Prospects and problems. PMID- 10462905 TI - CE marking and medical device directives. PMID- 10462906 TI - The IVD directive. PMID- 10462907 TI - Does the IVD directive apply to culture media? PMID- 10462908 TI - Control of substances hazardous to health: a continual process, not 'once for all'. PMID- 10462909 TI - [Dynamic correction of occlusal surfaces with CAD/CAM methods. Part I. General description of dental CAD/CAM-systems]. AB - The first part of the article presents a brief history and the present stand of dental CAD/CAM systems. The three basic components--scanning, design (CAD) and milling (CAM)--of such systems are described. Their advantages and disadvantages against conventional dental techniques are presented also. One of the most serious shortcomings of these "high-tech" systems is their impossibility to produce dynamic occlusal surfaces. PMID- 10462910 TI - [The use of Betadine antiseptic in the treatment of oral surgical, parodontological and oral mucosal diseases]. AB - The Betadine is an antiseptic PVP (polyvinil prrolidon)-iodine containing, water soluble solution, which has been in circulation in Hungary for years. Until now Betadine was not applied in the oral cavity for bacterial and fungal infections, although its effectiveness is unquery. The PVP-iodine is water solubilized that's why it does not sting the healthy or ill oral mucosa and has not side effects as discoloration of the teeth and tongue and trouble in the sensation of taste as does chlorhexidine. The only contraindication is iodine allergy. In the present study: in periodontal cases (abscesses, postoperative prevention) and in oral medicine cases alone or in combination with specific treatment schedule was Betadine applied. PVP-iodine as oral rinse was applied in 25 cases. According to our experience Betadine was excellent against bacterial and fungal infections in the oral cavity, which prevent or treat bacterial infections after surgical interventions. By this way the bacterial superinfections of exulcerant oral mucosal diseases avoidable. PMID- 10462911 TI - [Experience with clindamycin in stomatologic diseases]. AB - In 114 patients suffering from dental inflammatory diseases (periostitis, gingivitis, sinusitis, periodontitis etc.) and in 41 cases of dental surgical procedures (implantation, dentoalveolar surgery etc.) 3 x 300 mg/day clindamycin was applied during 7 days in the framework of GCP phases IV. In 150 cases (97%) the product proved to be effective and only in 2 cases a mild side effect was observed. On the base of the results clindamycin can be advised for the treatment of stomatological diseases. PMID- 10462912 TI - Image processing of collagen: role in medicine. AB - This article describes how quantitative studies of electron-optical images from collagen fibrils, used as a model system can provide information not only about the molecular architecture of the fibril but also about the structural alterations produced by a treatment or disorder. PMID- 10462913 TI - Side effects of lithium treatment on collagenous tissues. AB - Structural alterations of collagen fibrils induced by treatment with lithium chloride at various doses and durations, are investigated. In addition, this article describes how computer analyses of electron-optical images from collagen fibrils can lead to information about the changes produced by lithium on the molecular architecture of the fibril. PMID- 10462914 TI - Cytomegalovirus infection in infants: an example of a chronopharmacological approach. AB - A case of a male infant at the age of 3 months with cytomegalovirus (CMV) infection, diagnosed 40 days after the first symptoms, was discussed. While the right dosage and schedule during the initial treatment with ganciclovir (Cymevene) were agreed on, the right time of further application of anti-CMV IgG (Cytotest) and ganciclovir was unclear. Daily mean temperature was taken as an overall measure based on 7 single readings (6:00, 9:00, 12:00, 15:00, 18:00, 21:00 and 24:00 h +/- 10 min). Dynamics of daily mean temperature was studied by descriptive statistics, regression analysis, autocorrelation and periodogram regression analysis (software package "6-D Statistics" PC ver.4.5-98 by B. P. Komitov) with the purpose of identifying the time periods of viral DNA replications and CMV population growth inclinations. A five-step procedure was applied: (1) description of a tendency in the daily mean temperature; (2) studying variations in time series of the daily mean temperature; (3) decomposition of cyclic variations; (4) reconstruction of time series; (5) best model determination and forecasting. Three main cycles in variations of daily mean temperature were revealed (period T approximately 2.25-3.25, 5.5 and 12.25 days, p < 0.05) until the 37th day of therapy, when a strongly decreasing trend of daily mean temperature emerged. The cycle of 2.25-3.25 days disappeared and the daily mean temperature continued to decrease significantly on the background of improvement of the clinical status. It was concluded that (1) cyclic patterns in daily mean temperature (periods T = 2.25-3.25 and 12.25 days) during the etiologic medication stages could be due to the life-cycle of 2-4 days of the cytomegalovirus and a cycle in the viral population growth, respectively. The above findings confirmed previous results on the period of viral DNA replication from in vitro studies. It was possible to forecast the right moment of replication and viral load to adjust the treatment schemes and improve the outcome; (2) the complex time-series approach has shown to be very useful and effective in analyzing and forecasting the temporal dynamics of daily mean temperature in order to optimize the clinical management in this particular case of an infant with CMV infection. PMID- 10462915 TI - Ancient evidence for the origin and early development of chronobiology and biometeorology in Bulgaria. AB - Chronobiology and biometeorology have developed since ancient times of world history (China, Greece, etc.). Different authors have studied the origin and early development of medicine and related sciences in Bulgaria (Apostolov et al 1982, Georgiev 1987, etc.). They have reported that early medical postulates have incorporated the notion of temporal patterns of health and disease and their dependence on the environment. However, these authors have neither emphasized the above relationships nor considered them possible origins of both chronobiology and biometeorology in Bulgaria. The aim was to present evidence for the possible origin and early development of chronobiological and biometeorological perceptions in ancient Bulgaria. Different sources from ancient Bulgarian history were searched and analyzed. Ancient sources from Bulgarian history, relevant to the aim of this study, were reviewed. The evidence on chronobiological and biometeorological understandings in ancient times was obtained from the "Protobulgarian Calendar", Preslav's Rosette, etc. Here, only the most important of Protobulgarian, Thracian and Slavonic sources were presented. It was concluded that chronobiology and biometeorology had also their origin and early development in ancient Bulgarian times. PMID- 10462916 TI - Non-Hodgkin's lymphoma in US children: biometeorological approach. AB - Previous studies have shown cyclicity in the incidence of malignant melanoma of the skin (Houghton e.a. 1981, Dimitrov 1998, etc.) and associations of such variations with heliogeophysical activity (HGA). The purpose of this study was to investigate trends and patterns of variations in the incidence of non-Hodgkin's lymphoma (NHL) in US children (1973-1989) and their possible relationships to variations in HGA (Rz-index, aa-index). Linear and non-linear regression, autocorrelation, periodogram regression analysis and phase-correlation analysis were applied. No significant trend in incidence of NHL has been found but high frequency cyclic patterns of variations were revealed (period T = 4 years, p < 0.05). Similar cyclicity has been described also in HGA (period T = 5 divided by 6 years, p < 0.05). Lag-periods in associations of NHL incidence with Rz-index and aa-index have been described--dT = 6, 8, 10, 12 and 12 years, respectively (p < 0.05). To conclude, maximal rates of NHL incidence are most likely to appear on the descending slopes and about minima of HGA cycles, i.e. out of phase. Our results are consistent with the hypothesis of positive associations between incidence of non-Hodgkin's lymphoma and solar phenomena (Cartwright e.a. 1994, etc.) PMID- 10462917 TI - Possible mechanism of action of vanadium ions as an antidiabetic agent. AB - Vanadium compounds, at much higher concentrations than they are typically ingested, are being considered for use in the treatment of diabetes mellitus. They exert an insulin-mimetic effect in an insulin-receptor-independent manner. In our study we obtained new data about the vanadium insulin-receptor-independent mechanism of action on cell membranes. When rat stomach smooth muscle samples are treated with NH4VO3 (10(-7) divided by 10(-5) this action is possibly exhibited with increased influx of Ca2+ through VDCa2+C. PMID- 10462918 TI - Influence of odour immissions from cellulose-paper industry on some health indicators. AB - The population of the town of Stamboliisky, Bulgaria, is constantly exposed to air pollution which includes immissions of unpleasant odours like methylmercaptane, dimethylsulphide, hydrogen sulfide, etc. Most of these can cause serious olfactory damage even at concentrations well below the toxic levels. The aim of the present study was to analyse the effect of odour immissions on the health status of the people in the town of Stamboliisky. PMID- 10462919 TI - In vitro study on cytotoxic effect of some chemicals on serum McCoy and serum free McCoy-Plovdiv cell culture systems. AB - The cytotoxicity of test agents on serum-free McCoy cultures has not been studied at all. The cytotoxic effect of EDTA, methanol, DMSO, and cycloheximide on serum free McCoy-Plovdiv cell culture (SF) was detected visually on inverted microscope and quantitatively by tests for viability (NR) and total protein (KBP). The IC50 values for the tested chemicals were calculated. SF showed the lowest IC50 values for cycloheximide, DMSO and EDTA and the highest for methanol according to both tests. EDTA, methanol, DMSO and cycloheximide had dose-effect relationship in the cell test systems after treatment. The data indicate that McCoy-Plovdiv cell line is a suitable serum-free cell system for in vitro cytotoxic studies. PMID- 10462921 TI - Simple phantom for gamma camera spatial resolution objective measurement. AB - This paper presents a simply designed line source phantom for gamma camera spatial resolution measurement. It can be useful for the nuclear medicine departments where sophisticated spatial resolution measuring tools are not available; it can be used both in routine and in reference tests of gamma camera spatial resolution. PMID- 10462920 TI - Inhibiting effect of desalted extract from Galega officinalis L. on platelet aggregation. AB - The inhibiting and disaggregating effect of desalted and fractionated herbal extract of Galega officinalis L. on platelet aggregation in vitro is studied. At a concentration of 35 micrograms/ml in a platelet-rich plasma (PRP) the fraction inhibits 50% of aggregation by ADP and at 125 micrograms/ml PRP it inhibits fully the aggregation of PRP by ADP. At a concentration of 40 micrograms/ml PRP the fraction inhibits the initiation of platelet aggregation by collagen and at 50 micrograms/ml PRP inhibits the initiation of aggregation by thrombin. At a concentration of 65 micrograms/ml PRP the fraction can disaggregate 50% of the aggregated platelet-rich plasma by ADP and 25% of aggregated PRP by collagen. PMID- 10462922 TI - Time evaluation of the treatment of essential cyst bone in children. PMID- 10462923 TI - Survival of patients with surgery for lung cancer. AB - INTRODUCTION: Lung cancer is the leading cause of death in oncological diseases all over the world. It is a major health problems in Bulgaria. Early diagnosis is of great significance for improving survival in this group of patients. OBJECTIVE: The aim of this study was to analyze survival of patients with surgery for lung cancer. MATERIALS AND METHODS: Retrospective information for all 86 patients with surgery for lung cancer during the period 1995-1998 was obtained from the surgery protocols in the Clinic of Thoracic and Abdominal Surgery at the Higher Medical Institute (Plovdiv, Bulgaria). However, relevant follow-up data were found only for 50 patients. Medical histories were provided by the Plovdiv Oncological Center. Kaplan-Meier analysis was applied to study survival of patients. The log rank test was used to compare cumulative survival functions between groups of patients with different stage at diagnosis. Univariate Cox regression analysis allowed the development of a model to assess the influence of the stage at diagnosis on cumulative survival. RESULTS: Survival was assessed till June 1998. Mean survival period was 15 months (95% CI 13, 18). Patients were divided into two sub-groups: group A (stage I and II) and group B (stage III and IV). The analysis showed a marginally significant difference in cumulative survival between two groups (p = 0.0599). The mean survival period of patients from group A was 18 months (95% CI 14, 22) while for group B it was 12 months (95% CI 9, 16). Univariate Cox regression model showed that the relative risk of patients in group B to patients in group A is 3.2 (95% CI 1.11, 9.06). Early diagnosis of lung cancer is of crucial importance for surgery management and prognosis in such patients. PMID- 10462924 TI - Our experience in operative treatment of secondary and recurrent echinococcosis. AB - The frequency rate of echinococcis has recently increased rapidly and so has the number of the relapses of the disease. The aim of the present retrospective study was to summarise our experience in the surgical treatment of secondary and recurrent echinococcosis. We discuss the most effective methods for operative treatment in which the intervention spares organs and combines with further antiparasytic treatment. PMID- 10462925 TI - A case of abnormal localization of osteomyelitis. AB - A patient with osteomyelitis and abnormal localization of the changes is presented--left sterno-clavicular joint, the medial parts of the clavicular bone and the first rib, as well as the manubrium sterni. General and target radiography, tomography and chest CT-scan were performed. The reported case is of interest because of the rare localization, the characteristics of the X-ray conduct and the symptomatology in differential and diagnostic aspect. PMID- 10462926 TI - Effects of short-term exercise training in patients with heart failure. AB - Heart failure is a debilitating disorder which limits exercise capacity and produces a poor quality of life. The present study was designed to determine the effects of an exercise training program on patients with CHF NYHA functional class II-III, attributed to left ventricular systolic dysfunction and dilated left ventricle. Twenty-two ambulatory male patients with stable CHF were randomised to a training (n = 15) and a control group (n = 7). A symptom limited ramp cardiopulmonary exercise test with gas exchange analysis was performed at baseline after 4 and 8 weeks. The training group underwent an exercise training program at 50% of peak oxygen uptake for eight weeks. The control group was not exercised. After 8 weeks, compared with baseline, there were statistically significant increase in peak oxygen uptake, peak workload, anaerobic threshold, oxygen pulse, RPP, ventilation and the duration of the test only in trained patients. The reduction in scores tested by the Minnesota Living with Heart Failure questionnaire (p < 0.001) and Borg dyspnea rating score (p < 0.001) reflect the reduction of symptoms and the improvement in health-related quality of life. Carefully selected patients with moderate to severe CHF can achieve significant improvements of exercise capacity and quality of life with exercise training and can safely participate in a conditioning program. PMID- 10462927 TI - Diagnostic strategies for evaluation and prognosticating the outcome of jaundice among patients with cholestasis caused by neoplastic diseases of the hepatobiliary system and the pancreas. AB - The purpose of the present study was to assess the diagnostic approach and the outcome of neoplastic diseases of the hepatobiliary system and the pancreas that develop cholestasis. 84 patients over 40 years of age treated in our Department between January 1994 and January 1998 were included. The etiological cause of jaundice was established by ultrasonography, computed tomography, thin needle biopsy with cytological and histological examination of the obtained material or ERCP. 11 of the 84 patients died, 9 underwent surgery and 64 were referred to the Oncologic Center for further treatment. PMID- 10462928 TI - Multichannel alternate electrostimulation using the new Bulgarian Vita 2007 equipment in post-stroke rehabilitation. AB - In this study the authors evaluated the effect of a new method--multichannel alternate electrostimulation using the new Bulgarian equipment Vita 2007 for regulating muscular imbalance, breaking the pathological synergic patterns and overcoming motor impairment after stroke. The subjects of the study were 15 patients with hemiparesis secondary to stroke. The beneficial results in accelerating motor recovery and assisting the physical exercise programme for recreating proper patterns of walking and manipulative activity were assigned to the change in the level of spasticity and to the new method of consecutive alternate stimulation of the muscles that take part in the normal movement. PMID- 10462929 TI - Study on proinflammatory cytokines (IL-1 beta, IL-6, TNF-alpha) and IL-2 in patients with acute hepatitis B. AB - Cytokine production in patients with hepatitis B may be related either to multiple immune abnormalities or to favourable outcome of the disease. Serum levels of IL-1 beta, IL-2, IL-6 and TNF-alpha were determined dynamically by ELISA kits in patients with self-limited form of acute hepatitis B infection (A HBV) during the first, second and third decade after icterus appearance. IL-1 beta, IL-6 and TNF-alpha had characteristically high levels in patients measured in all decades, but these were the highest during the first. Then they gradually decreased, reaching normal values at the stage of convalescence. Serum IL-2 levels were found to be most significantly elevated during the second decade and also dropped to normal values in the course of the disease. Patients who cleared HBsAg on the third month after dehospitalisation had higher mean values of IL-2, IL-6 and TNF-alpha in comparison with those who were still HBsAg carriers, thus indicating that proinflammatory cytokine production in self-limited HBV may be important for viral clearance. PMID- 10462930 TI - Neonatal transport--a part of the study "follow-up of some indices in neonates, treated with artificial ventilation during the neonatal period". AB - Surveillance of a sick neonate is closely related to appropriate intensive care and therapy in neonatal intensive care unit (NICU). There will be babies that need to be transferred to NICU. We used the neonatal intensive computer file (NICF) as an information base. Our results showed clear-cut differences between the studied parameters both for the different BW groups, survived/nonsurvived and transported/nontransported groups: differences in sex, Apgar score, number of neonates in 1992. tendency towards earlier transport for VLBW, followed by LBW and TB. the longest period of IPPV + IMV/weaning from ventilator in transported LBWS. severe disturbances, diseases and malformations noted in nonsurvived both transported and nontransported. PMID- 10462931 TI - Treatment of upper respiratory tract infections in the first year of life without antibiotics and antipyretics. AB - There is still no consensus on the question whether it is necessary to use prophylactic antibiotic therapy and on the febrile states in infectious diseases. The present study aims at solving these problems. Between 1996 and 1997, 128 children under one year of age with upper respiratory tract infections (URTI) were hospitalized in the department and examined using clinical, laboratory and instrumental methods. We found no significant difference in the percentage of complications in the children treated with or without antibiotics. We conclude that prophylactic antibiotic therapy is not justified for the predominating viral etiology of URTI in children less than one year old. URTIs can be managed without antipyretics, which entails no risk for the patients. PMID- 10462932 TI - Application of low intensity laser radiation in extravasal infiltrates caused by cytostatics in children with acute leukemia. AB - Application of low intensity laser radiation in the field of infantile oncohematology has not been discussed in the available medical literature. The possible adverse effect of the physical factors on the main disease and the controversial character of their application prevent their being used in the treatment of these diseases. The aim of the present study was to assess the feasibility of using low intensity laser radiation (LILR) in treating extravasal infiltrates caused by cytostatics (vincristin, daunorubicin) administered in children with acute leukaemia (AL). We applied laser therapy in a group of children with AL in the stage of exacerbation. The children were undergoing chemotherapy with cytostatics. Immediately after formation of the extravasal infiltrates the constant leaders in the venous duct were removed and low intensity laser therapy with LMS "Prometheus" was administered. This apparatus is a semi-conducting soft laser in the infrared spectrum (890-910 nm wavelength, depth of penetration 25-30 mm) working in an impulse mode. We used minimal doses in the range of the anti-inflammatory enthrophic frequencies. We found that the extravasal infiltrates rapidly decreased the local inflammatory response and the pain was relieved. The strong anti-inflammatory effect can be accounted for by the nonspecific action of the LIRR on the tissues, which is manifested by better microcirculation, removal of toxic substances and stimulated regeneration capabilities of the cells. This study is an attempt to work successfully in this very delicate field of the contemporary medicine despite the abstention from using it before. PMID- 10462933 TI - Analysis of the results from an early low-level predischarge and late symptom limited exercise tests and coronary angiographic examination in patients with uncomplicated myocardial infarction. AB - The aim of our study was to evaluate the prognostic value of an early predischarge and late symptom-limited exercise tests as opposed to the evidence from the coronary angiographic examination. Twenty patients with an uncomplicated myocardial infarction were studied with an early predischarge exercise test (14 21 days) and a following postdischarge late symptom-limited veloergometric test. All the patients underwent an exercise test designed according to the protocol of the International Health Organization. A coronary angiographic examination was carried out in all the studied patients. Thirteen of the patients were with two positive veloergometric tests and significant stenosis from the selective coronary angiography. Seven of the patients had two negative tests. Two of them did not have significant coronary stenosis from the angiografic assessment. Generally 40% of the patients undergoing late exercise test reached a higher workload. There was no significant difference between the results from an early predischarge and late symptom-limited veloergometric test. A higher physical work load reached during the late exercise test did not show an improved coronary blood flow. Positive early and late exercise tolerance tests proved significant coronary stenosis. A negative exercise test did not exclude significant coronary stenosis. PMID- 10462934 TI - Integrative metabolic effect of risk factors and the pathogenesis of insulin resistance. (theoretic speculations and therapeutic corollaries). AB - This paper analyses theoretically insulin post-receptor resistance caused by risk factors (RFs)--obesity, energy over-load, low physical activity, stress. Each of these RFs is capable of producing abnormalities directly or indirectly in glucose and lipid metabolism solely or in combination with the others. The RFs pathogenic integration forms in each patient an individual kind of pathogenic integral (PI) which becomes pathogenic because of RFs' pathogenic power summation. Sulfonylurea therapy may increase insulin resistance and become a part of PI. Waiting for perfect medicine, the intensive clinical treatment of RFs seems to be the most reasonable behavior in NIDD. PMID- 10462935 TI - Statistical report from Uludag Poison Information Center--34-month experience. AB - Management of poisoned patient requires accurate knowledge of intoxication. Uludao Poison Information Center (UPIC), founded in January 1996, provides acquired information 24 hours a day 7 days a week for physicians and consultants. We received 2270 phone calls since January 1996 till October 1998 and evaluated the cases statistically regarding "distribution of age and gender", "calling time after poison exposure", "source of poisoning", "reason for intoxication" and "source of calls" to reveal the distinguishing features of intoxication. Intoxication was mostly seen between 14-25 years of age (964 cases, 42.46%) and in females (1367 cases, 60.22%). Drugs (1400 cases, 61.67%) were the main source and suicide attempts (930 cases, 40.9%) were the main reason of poisoning. Most of the calls were from healthcare professionals (1347, 59.33%). PMID- 10462936 TI - Psychosocial problems in elderly subjects from geriatric home revitalization approach. AB - The psychosocial aspects of aging and senility appear a major problem for the marked senescence of the Bulgarian nation--25% of the population is over the age of 60 years. The present study attempts to make a medico-ethic and social evaluation of life quality in old age and illness and outline possibilities for appropriate revitalization of elderly subjects dwelling geriatric homes. The governing body of the geriatric home does not refer to scientific methods of management for planning and organization of everyday living activities. No coping strategy is applied to reduce stress and illness offence. Current scientific revitalization approach should be realized in the policy of the geriatric home to prevent a long-term disabled existence and induce a worthier lifestyle. PMID- 10462937 TI - Age at marriage and intrafamily planning in young families. AB - INTRODUCTION: The sociological inquiry on current problems of intrafamily planning carried out in 1995 provided opportunities for finding the effect of a number of social and demographic factors on the formation of family. OBJECTIVE: The present study has investigated the influence of the age of woman at marriage on the main indicators characterizing the reproductive behavior. MATERIAL AND METHODS: Direct individual inquiry was performed and 500 young families from the town of Plovdiv were approached. Data were analyzed by percentage distribution methods and non-parametric tests. RESULTS AND DISCUSSION: We should note here the high percentage of spouses who had not planned the number of children in their future family--33.1% of women and 34.3% of men. Paradoxically, the percentage of women that had not planned the number of children in their families increases with age. The intended number of children does not depend on the age of women at marriage. The women aged below 19 have indicated the highest number of children. The "ideal number of children" indicator has the highest value in women aged below 19 (2.25 children), but there is no statistically significant difference in the ideal number of children between the different age groups. The opinion about a "large family" is studied in spouses and relatives of the young family. With the increase of age, the number of children in the "large family" decreases in all three groups (P < 0.001). There is no difference in length of the protogenetic interval between different age groups. Intergenetic interval between the first and second birth is about three years in all three age groups. The largest interval between the second and third birth is registered in women aged above 25. About half of the women have admitted that they do not wish to have a third child. The age at last delivery is highly correlated with age at marriage (P < 0.05); the correlation is strongest in women above 25 years of age. PMID- 10462938 TI - Estimation of trends of morbidity of colorectal cancer in Plovdiv district for the period from 1985 to 1996. AB - The present study concerns the incidence rate of colorectal cancer (CRC) in Plovdiv region (population 1,269,464). The study was carried out over a 12-year period and aimed at establishing the increase rate of this disease. Results show that incidence rate of CRC increased progressively for the mentioned period from 22.14 to 34.98 per 100,000 people and was almost twice as high as that for the country. It is a particularly disturbing fact that the morbidity and mortality rates due to colorectal cancer is rising--2.36 times for 12 years. We propose screening for asymptomatic and high risk patients with considerable results for opportune diagnostics. PMID- 10462939 TI - Influence of family tradition on reproductive behavior of Gypsy population. AB - INTRODUCTION: Stability of traditional opinion in different ethnic communities determines to a great degree the demographic behavior of their members. AIM: To analyze the influence of family traditions and system of values on the reproduction of gypsy population. MATERIALS AND METHODS: The opinions of 495 Gypsy women in reproductive age was studied by direct individual questionnaire. The answers characterizing the reproductive behavior of two successive generations were analyzed. The following statistical analyzes were applied: descriptive statistics, chi-square and t-tests. All p-values are two-tailed. The statistical analyses were made on a PC using SPSS 5.0.2. RESULTS: Mean age at marriage in different generations under study was similar (it is 15.50 +/- 0.10 years in the present generation vs. 15.58 +/- 0.09 years in the previous generation, p > 0.05). The same is valid for the mean age at the first birth (16.98 +/- 0.11 years vs. 17.04 +/- 0.10 years, respectively, p > 0.05). However, there was a statistically significant difference in the mean number of live births between the generations (2.37 +/- 0.05 vs. 4.10 +/- 0.09, respectively, p < 0.001). The latter finding might be explained by fact that the women from the present generation are still in their reproductive period. CONCLUSION: It is obvious that family traditions may have, to a certain extent, an influence on the patterns of reproductive behavior in gypsy population. These traditions represent an indispensable part of their lifestyle and their intentions to have many children. PMID- 10462940 TI - Malignant melanoma of the skin and non-Hodgkin's lymphoma in USA: a comparative epidemiological study. AB - Possible associations of incidence of non-Hodgkin's lymphoma (NHL) with solar UVR have been suggested (Cartwright e.a. 1994, etc.). Also, incidence for both malignant melanoma of the skin (SMM) and NHL has shown a rapid increase over the last 10-15 years worldwide. On the other hand, cyclicity in variations of incidence rates for SMM was reported (Houghton e.a. 1981, Dimitrov 1998, etc.). The present study analysed trends and variations in incidence of SMM and NHL for the USA. Annual age-standardized incidence rates for SMM and NHL (SEER database, 1973-1989) were compared. Linear and non-linear regression modeling, periodogram regression analysis and parametric tests were applied (Dimitrov e.a. 1995). The analysis denoted non-linear trends of increasing incidence for both SMM and NHL. Cyclic variations in incidence rates for SMM in US females were reported previously (Dimitrov 1993) and now were confirmed also for other strata of the population (mainly, cycles with a period T = 8 divided by 9 years, p < 0.05) as well as revealed for NHL (mainly, cycles with a period T = 8 divided by 9 years). This cyclicity was significant at 95% only for black US males (Table 1). After decycling for hypercyclicity of 20-35 years (long-term cyclic trends), however, significant high-frequency infrannual variations of 3 divided by 5 years appeared in NHL (latter cyclic variations, for instance, were observed in the sunspots index, solar UVR and stratospheric ozone). Also, significant cycles of 11-13 years appeared for white US males. Similar cyclicity of 9 divided by 11 years was observed in sunspots number over the same interval. Trends of NHL were very similar to that of SMM, as were the variations. This study might be considered a contribution to the hypothesis on possible associations of incidence for NHL with heliogeophysical phenomena. PMID- 10462941 TI - Non-pharmacological management of the behaviour of pediatric dental patients. AB - Behaviour modelling is frequently used to modify children's behaviour. The psychological techniques of encouragement-reprobation are an integral part of the behaviour shaping. Three hundred clinically healthy children were recruited in this study. They were aged 54-96 months and allocated to three groups according to the specific technique used: group 1 in which we applied the "live patients model" technique, in group 2 the "encouragement-reprobation" techniques was applied and group 3 was a control group. The patient's behaviour was assessed using L. Venham's Cooperative Behavioral Scale. A behavioral improvement was noticed in the experimental groups after applying the techniques for behaviour modification. The comparison shows a statistically significant difference between the two experimental groups and the control one and absence of a significant difference between the influenced groups. The study shows that there is a stable for behaviour improvement. PMID- 10462942 TI - Delayed diagnosis of cancer with emphasis on oral cavity cancers. AB - Delayed diagnosis of cancer and, of oral cavity cancers in particular, is of crucial importance for the clinical management, costs of care and unfavourable outcome. Many scientists have studied the causes of delayed diagnosis to find solution of the problem. However, only few studies of this kind have been performed in Bulgaria, especially for oral cavity cancers. The aim of this overview was to summarize and analyze publications on problems and causes of delayed diagnosis of cancer in general and, of oral cavity cancers in particular. The literature search was made in different databases such as MEDLINE, BIOSIS, INTERNET, etc. More than 100 relevant items of information were extracted but only 22 of them were found suitable and further analyzed. The analysis of the literature has revealed the importance of recognizing the warning signals and causes of delayed diagnosis in cancer in general and in oral cavity cancers in particular. For instance, survival of patients with squamous cell carcinoma of the tongue (Dx:141) was related to delayed diagnosis (20% of patients survive more than 5 years and 71%--only 2 years after diagnosis). The problems of delayed diagnosis in patients with multiple neoplasm of the oral cavity (Dx:143-145) and with nasopharyngeal carcinoma (Dx:146-149) were also discussed (96.6% of patients were in stage III at diagnosis). This overview has summarized problems in and stressed the main aspects of delayed diagnosis of cancer in general. It has also denoted hints in delayed diagnosis of cancer of the oral cavity and their relevance to the clinical management, outcome and costs of care for such patients. The latter has been considered of primary importance for general practitioners, dentists and specialists in public health medicine. PMID- 10462943 TI - Measuring the quantity of phosphor in the saliva after application of fluoride gel. AB - The transition of active ingredients from the prophylactic agents into the saliva leads to an increase of its mineralizing potential. The aim of the present study was to measure the quantity of phosphor in the saliva after application of a gel containing both fluoride and phosphate ions. The quantity of phosphor is measured photometrically according to the Chen method. Results show a sharp increase in the quantity of phosphor in the saliva after application. It gradually decreases with time and after 2 hours it reaches the initial level. PMID- 10462944 TI - Treatment of postsurgical jaw bone defects by bone filling material. AB - The aim of this study was to analyse the clinical application of some bone filling materials (DFDBA, Osteovit, ATR-24) during the treatment of limited postsurgical defects of jaw bones in 108 patients. The results indicated a significant decrease of the mean time needed for bone regeneration within the area of the defect as assessed by Ro-densitometry and videodensitometry during the follow-up period. PMID- 10462945 TI - Calculating the force tension at horizontal pressure of teeth and different degree of decomposition of parodontium. AB - The aim of this study was to analyze the relationship of decomposition of parodontium at applied horizontal pressure on teeth. A model tooth has been constructed which allows quick and precise calculating of force tensions. The results have shown that increased force tensions lead to the destruction of the parodontium and can be used for planning of prosthetic constructions. PMID- 10462946 TI - Treatment of bruxism and bruxomania (clinically tested). PMID- 10462947 TI - The combined IGG, IGM and IGA peroxidase conjugate can facilitate determination of immune complexes by CIF-ELISA and anti-C3 ELISA. AB - To determine the levels of circulating immune complexes (CIC) in normal and patients sera, CIF-ELISA and anti-C3 ELISA were performed. Immune complexes containing different antibody isotypes were detected simultaneously by the combined anti human IgG, IgM and IgA peroxidase conjugate as detecting antibody. The results obtained confirm the higher CIF-ELISA sensitivity, specificity and reproductivity compared to anti-C3 ELISA and provide good evidence to justify the use of CIF-ELISA as a screening test for CIC assessment. PMID- 10462948 TI - In vivo testing of Enterogenin for a mutagenic activity of bone marrow hemopoietic tissue of mice. AB - The purpose of the present study was to test Enterogenin for genotoxicity. The micronuclearic test of Schmid W. was used. Enterogenin was introduced intraperitoneally, once and/or twice in two doses on male and female mice of the BDF 1 hybrid line. The positive controls were injected with clastogenic agent. Bone marrow smear preparations were made, determining the ratio between the normochromic erythrocytes (NCE) and polychromic erythrocytes (PCE), and the number of micronuclearic erythrocytes (MNE). Enterogenin has no genotoxic effect on the hemopoietic tissue of bone marrow of mice the ratio NCE/PCE being within the reference values. The preparation showed no clastogenic effect (p > 0.05), compared to the controls, as the number of PCE with micronuclei did not change 24, 48 and 72 hours after treatment with the two studied doses. PMID- 10462949 TI - Functional parameters in pulmonary bullous emphysema. AB - OBJECTIVE: 1. compare a large set of functional parameters in patients with bullous emphysema and patients with nonbullous emphysema. 2. To compare a chest radiographs (CHR) and a high resolution computed tomography (HRCT) in the clinical assessment of bullous emphysema. MATERIAL AND METHODS: The study population included 43 stable COPD patients (age = 59 +/- 9 years; pack/year (P/Y) = 39 +/- 19; ATS dyspnea score = 2.3 +/- 0.9; FEV1%pred. = 30 +/- = 10%; KCO%pred. = 49 +/- 16%; 6MWD (six minute walk distance) = 395 +/- 103 m; mean +/- SD). The patients were divided into two groups (patients with and without bullae) by a HRCT. In most of the cases the size of the bullae, measured by CT scan, was less than 15 mm. Twenty two CHRs were read independently by three experienced chest radiologists who had no knowledge of the CT scan data. RESULTS: Statistically significant differences were found between the groups with bullous (n = 19) and nonbullous (n = 24) emphysema in FEV1 (p < 0.001); VC (p = 0.001); BMI (p = 0.018); Borg after exercise (p = 0.021); FEV1/VC% (p = 0.025) and P/Y (p = 0.034). The sensitivity of chest radiographs compared with CT scan regarding the small bullae was very low: 27.7% in radiologist I, 12.3% in radiologist II, and 21.5% in radiologist III. CONCLUSIONS: 1. The patients with bullous emphysema have statistically significant lower lung function indices (FEV1, VC, FEV1/VC%) and BMI than those with nonbullous emphysema. 2. Patients with bullous emphysema have higher level of dyspnea score after 6MWD and higher pack-year smoking status than those with nonbullous emphysema. 3. For the clinical evaluation of emphysema the information derived from a standardised reading of the CXR is not as valuable as that derived from the CT scan. PMID- 10462950 TI - Study of suicidal risk and tendency in defectologists and logopedists from a school for handicapped children. AB - This study was carried out to ascertain the suicidal risk and tendency among 26 defectologists and 3 logopedists. The objective was to find the percentage of teachers working in the specific and hard conditions of a teaching process among whom suicide tendencies or high suicidal risks are observed. PMID- 10462951 TI - Prognostic significance of the immunological variant in patients with multiple myeloma. AB - INTRODUCTION: The prognostic significance of the immunological variant and especially of the type of light chain secretion in multiple myeloma is controversially assessed in the literature. OBJECTIVE: To study the prognostic significance of the class and type of the monoclonal production in patients with multiple myeloma. PATIENTS AND METHODS: 109 patients with multiple myeloma were enrolled, in the study, diagnosed and treated at the University Clinic. The class and type of the paraprotein secretion was determined using immune electrophoresis of serum and/or urine by Grabar and Williams, Mancini radial immunodiffusion and Ouchterlony immunodiffusion for light chain typing. The median survival (MS) was calculated using the method of maximal logarithmic authenticity, where the survival in months = log t. RESULTS: The MS of all patients was 29 months. Patients with Bence Jones myelomas showed the lowest survival--12 months; followed by those with IgD myelomas--20 months. The patients with combined and non-combined light chain production (BJ+) lived 26 months, with BJ(-)--35 months (inauthentic difference). No difference was found between the survival of BJ(kappa)--30 months and BJ(lambda)--21 months. BJ(+) patients with myeloma nephropathy and nitrogen retention had a MS of 23 and 20 months, respectively, as compared with the BJ(+) patients without myeloma nephropathy and nitrogen retention: 47 and 40 months (p < 0.05). DISCUSSION AND CONCLUSION: The class of the paraprotein secretion only and the two types of light chain separately do not possess any prognostic significance. From a quantitative point of view the class and type of paraprotein production are independent staging criteria with certain predictive value. The prognostic significance of BJ-proteinuria is found making comparison between the groups in which its pathogenic effect is expressed. PMID- 10462952 TI - [Surgical treatment in child gynecology. Germ cell tumors of the ovary in girls]. AB - The authors presented 50 girls with germ cell tumours of the ovary. 9/50 (4.5%) were malignant germ-cell tumours. There were 5 patients with pure dysgerminomas, 1 with endotermal simus tumor, 3 with immature teratomas, 3 with gonadoblastoma, 38 with mature teratoma. Kariotype 46XY was present in patients. Authors prefer fertility sparing operative treatment. Primary postoperative therapy in malignant tumors was chemotherapy or radiotherapy. Authors recommend sift out ultrasonographic investigation. Girls should be individually considered for appropriate ovarian surgical procedures. The operative treatment should be as conservative as possible. Absence of follicle apparatus in the gonads require determination of levels of gonadotropin and kariotype. PMID- 10462953 TI - [Ovarian cancer in girls]. AB - DESIGN: The aim of our study was a clinical analysys of diagnosis, operative treatment and monitoring of epithelial ovarian cancer in girls. MATERIALS AND METHODS: Authors described 11 epithelial malignant tumors of ovaries in girls aged 12-16. Clinical aspects, pathology stage, grade treatment, monitoring and survival were evaluated. Three of the patients had ovarian cancer. A 19-year old patient with stage Ia of epithelial ovarian cancer was managed conservatively. Four years later she give birth. For 10 years no malignant of has been observed. Seven patients with ovarian borderline epithelial tumors were treated. The tumors in 5 patients were of serous and in 2 of mucinous type. CONCLUSIONS: We suggested that patients of stage I who wish to preserve childbearing function may be treated with unilateral salpingo-ovariectomy and chemotherapy. PMID- 10462954 TI - [The comparison of CA-125 and CEA levels in serum and ovarian cyst fluid in girls and young women]. AB - OBJECTIVES: The aim of the study was to compare the CA-125 and CEA levels in serum and ovarian cyst fluid. MATERIALS AND METHODS: Concentration of the CA-125 and CEA were measured at 12 girls and young women with benign, low malignant potential and functional cysts hospitalized in our teaching department. Concentrations were measured using immunoenzymatic method. RESULTS: We did not observe any difference in serum levels of CA-125 and CEA between neoplastic and functional cysts. On the contrary we found very high levels of the markers in the neoplastic cyst fluid in comparison with the functional ovarian cyst fluid. CONCLUSION: It seems that high levels of the CA-125 and CEA in neoplastic cyst fluid in comparison with functional may be useful in differential diagnosis. PMID- 10462955 TI - [The levels of selected hormones in serum and ovarian cyst fluid in girls and young women]. AB - OBJECTIVES: The aim of the study was to determinate levels of the FSH, LH and E2 in serum and ovarian cyst fluid in neoplastic tumors (7 benign, 1 of low malignant potential) and functional cysts. MATERIAL AND METHODS: We measured the concentrations of the FSH, LH and E2 at 12 girls and young women hospitalized in our teaching department using immunoenzymatic method. RESULTS: We found higher levels of gonadotropins in serum and particular in ovarian cyst fluid at neoplastic tumors in comparison with functional cysts. There were also high levels of estradiol and low of gonadotropins in functional ovarian cyst fluid. CONCLUSION: The differences of FSH, LH and E2 levels in ovarian cyst fluid in neoplastic and functional cyst have cognitive character and need more investigations. PMID- 10462956 TI - [Ovarian gonadal tumors in materials of the clinic of gynecological endocrinology of the Medical Academy in Poznan (1970-1998)]. AB - 11 patients with ovarian gonadal tumors were treated at Clinic of Endocrinological-Gynecology of Academy of Medicine in Poznan. All presented symptoms of hormonal ovetproduction. In 9 cases we performed adnexectomy, only in 1 case (patient after menopause) hysterectomy with bilateral adnexectomy. PMID- 10462957 TI - [Surgical approach in cases of adnexal pathology in adolescents and young women]. AB - Retrospective analysis comprised 108 cases of adnexal pathology found at laparotomy in the group of adolescents and young women up to the age of 25. The most common condition necessitating the laparotomy was hemorrhagic corpus luteum (24% of cases). Adnexal cysts of different but benign histological structure had been found in 30.5% of cases. Malignant germ cell tumors (6 cases--dysgerminoma and teratoma immaturum) all of which at Ia stage at the time of laparotomy, had been treated conservatively with unilateral salpingo-oophorectomy. Authors suggest that the conservative approach should be applied in most cases of adolescents' adnexal pathology. PMID- 10462958 TI - [Surgical treatment of ovarian tumors in girls in the materials of the 2nd Department and Gynecological Clinic of the Medical Academy in Wroclaw]. AB - Between 1987 and 1998 44 young women (average age 17.4) were operated for ovarian tumours at the 2nd Clinic and Department of Academy of Medicine in Wroclaw. Bilateral changes were observed in 6 patients. In one case a malignant tumour was diagnosed. The most frequent benign tumours were teratoma adultum (30%) and cystis paraovarialis (22%) the type of surgical intervention depended on the size of the tumour and on the degree of destruction of the ovarian tissue. Since 1995 the clinic carries out surgical laparoscopy particularly prefened modality in case of young women. During that time, 16 of 21 patients aged up to 21, had their ovarian tumours enucleated by means of the laparoscopic method. PMID- 10462959 TI - [Laparoscopy in the diagnosis and treatment of pathology of sexual organs in girls]. AB - DESIGN: The aim of our study was a clinical analysis of laparoscopies in girls which were performed in the Clinic of Gynaecological Surgery. MATERIAL AND METHODS: Laparoscopic procedures were carried out between 1996-1998. Patients were divided in to three groups: aged under 15, between 15-19 and over 19. 44% of patients from the youngest group had laparotomy. 43.8% of laparoscopic interventions were in aged group 20-24. CONCLUSION: Laparoscopy is very good diagnostic method in gynaecological diseases. It gave enabled operative treatment in 40-43.8% girls. PMID- 10462960 TI - [Laparoscopic evaluation of adolescent girls suffering from algomenorrhea and acyclic pelvic pain]. AB - 120 adolescent girl suffered from acyclic pelvic pain and dysmenorrhoe (algomenorrhoee) was presented. They were treated with nonsteroidal anti inflammatory drugs (NSAIDs) and oral contraceptive pills (OCPs) and underwent a laparoscopy to determine the etiology of pelvic pain. Endometriosis was classified according to American Fertility Society classification. Adolescent with pelvic pain not responding to therapy with NSAIDs and OCPs have different rate of endometriosis. PMID- 10462961 TI - [Endometriosis in adolescents and young women. Can we afford to delay the diagnosis?]. AB - Retrospective analysis performed on medical records of 129 adolescents and young women treated surgically at I Dept. of Surgical Gynaecology during the nine years period (1989-1998) revealed 22 cases (17%) of endometriosis. Main indication for surgical intervention, apart from dysmenorrhoea and cyclic abdominal pain, had been the presence of endometriod cyst, as disclosed by ultrasound investigation (66%). Despite of negative result of imaging or clinical examination subjective complaints necessitated laparotomy in 27% of cases. According to American Fertility Society endometriosis classification, 17% subjects presented endometriosis in I stage, 17%--in stage II and remaining 66%--endometrioid cysts with stage III. Authors conclude, that in cases with persistent abdominal pain, not responding to conventional therapy, diagnostic laparoscopy/laparotomy in young women should not be delayed, unless endometriosis is suspected. PMID- 10462962 TI - [Surgical treatment of polycystic ovary syndrome]. AB - This analysis includes 67 cases of diagnosed polycystic ovary syndrome, which were treated by surgical procedure (ovarian wedge resection). Subjects to assessment were: efficacy of treatment by mean of menstrual cycle regulation and influence of wedge resection on patients hormonal profile. PMID- 10462963 TI - [Operative treatment of developmental abnormalities of sexual organs in girls]. AB - DESIGN: The aim of our study was a clinical analysis of operative treatment of developmental abnormalities of sexual organs. MATERIALS AND METHODS: The authors describe developmental abnormalities of sexual organs in girls aged 12-19 79(1.4%) girls were treated from among 5432 surgically treated patients in the course 6 years. Among these abnormalities were the following: agenesia of vagina, agenesia of cervical canal, absence of cervix of uterus, obstructed normal or rudimental vagina, occlusion of hymen. There were complex abnormalities too. Congenital abnormalities were subject to the reconstructive operative treatment. CONCLUSIONS: We recommend that developmental defects should be treated in specialist centres well equipped and experience of in operative procedures and postoperative monitoring. PMID- 10462964 TI - [Laparoscopic modification of Vecchietti operation in the treatment of congenital vaginal aplasia]. AB - Four young women aged 16-18 years were treated by laparoscopic modification of Vecchietti operation because of congenital vaginal aplasia. In all cases the postoperative period was uneventful and in every case functional vagina with the length 12 cm was achieved within 4 to 14 days of treatment. No pain killers were used during the olive traction in order to create a vagina. Our own instrumentarium necessary to perform such kind of operation was presented. Technical aspects of laparoscopic modification of Vecchietti operation were discussed. PMID- 10462965 TI - [Primary amenorrhea caused by impaired cervical development: report of 3 cases]. AB - We report 3 cases of primary amenorrhea accompanied by cyclic abdominal pain. In presented cases primary amenorrhea was caused by the absence of normally developed cervix. We applied surgical treatment--hysterectomy. In 1 case before hysterectomy we tried to reconstruct cervical canal. This effort was unsuccessfull. PMID- 10462966 TI - [The case of uterus unicornis with rudimentary horn in a 15-year-old girl]. AB - The 15 years old adolescent with a rare mullerian anomaly, uterus unicornis with a rudimentary horn is presented. Patient was 15 years old girl suffered from pelvic pain. In this case we illustrate diagnosis, symptoms and therapy. Patient was submitted complex surgical treatment. PMID- 10462968 TI - [Giant bilateral ovarian tumor in a 14-year-old girl]. AB - We present the case of a 14-year old girl, who underwent surgical treatment at the ward in 1993 due to giant twisted bilateral ovarian tumours (cystes folliculares). She had been admitted to hospital because of pains located in the hypogastrium area, nausea and vomiting. Increased abdomen circumference had been noted. At the time of being admitted to hospital the girl was having menarche. Having considered both the course of the disease and the clinical and ultrasonographic examination results, surgical treatment was prescribed. It was decided that both adnexa should be removed. On the right side a tiny fragment of regular ovary parenchyma was left. After the surgery the girl did not menstruate. As a result of a few months' hormonal treatment the girl's menstruation processes were brought to a regular state. At the moment the patient is a 20-year old, properly developed young woman. PMID- 10462967 TI - [Surgical treatment in a case of partial vaginal obliteration coexisting with hematometra, hematocolpos and other congenital defects]. AB - A case history of 12 years old girl with transverse vaginal septum occurring with hematometra, hematocolpos and other congenital defects is presented. Operation was carried out twice because of vaginal prosthetic appliance after the first operation. The reoperation was done 6 months after the previous one and artificial vagina recanalised. PMID- 10462969 TI - [A case of coexistence of polycystic ovary syndrome and ovarian carcinoma]. AB - There was a case of carcinoma of the ovary that coexisted with PCO syndrome in young, 25 years old woman. After clinical examination and USG doctors suspected PCO syndrome and hydrosalpings that is why the patient underwent an operation. In intraoperative histopathological examination bilateral adenocarcinoma were found. In prophylactic examination of patient's mother ovarian tumour was found that is also turned out to be an adenocarcinoma. PMID- 10462970 TI - [Gestational trophoblastic disease in a 15-year-old girl]. AB - Authors described gestational trophoblastic disease in 15 year old girl with concomitant disseminated intravascular clotting. beta HCG was higher that 100,000 mIU/ml. Serum beta HCG level decreased after evacuation of a molar pregnancy. Physiological level was achieved 3 months later. Theco-lutein cyst grew smaller in the same time. PMID- 10462971 TI - [Androblastoma in a female patient with the signs of virility: a clinical morphological and genetic study]. AB - We report a case of 24 year old patient with secondary amenorrhea, virilization and elevated serum testosterone concentration, in which cystic-solid tumor in right ovary was found. Surgical treatment was applied--adnexectomy. Histopathologic examination supported suspicion of androblastoma. After treatment regulation of menses and serum testosterone level was obtained. PMID- 10462972 TI - [Breast malignancy in girls]. AB - The authors described 19 year old patient with breast cancer and two teen-age patients with juvenile papillomatosis of the breast. Authors have postulated that among different problems of pediatric gynecology it is necessary to notice pathology of breast. PMID- 10462973 TI - [Molecular background of hyperandrogenism of ovarian origin]. AB - Ovarian androgens constitute substrates for estrogen formation, the key enzyme in androgen production being 17 alpha-hydroxylase/17,20 lyase, while in estrogen synthesis, aromatase. Both enzymes are constituents of a complex, comprising two proteins: cytochrome P450 reductase and respective cytochromes P450c17 or P450arom, whose expression play an important role in a proper function of the ovaries and its genetically determined abnormalities may lead to appearance of clinical symptoms. Although it is difficult to clearly define the genetic background of disturbances, leading to hyperandrogenism, metabolic abnormalities, resulting in excessive androgen production, were observed in polycystic ovarian syndrome, hyperthecosis and hyperinsulinemia. In this review data from literature were presented and discussed regarding endocrine and molecular background of hyperandrogenism of ovarian origin. PMID- 10462974 TI - [The influence of IGF-1 on the biosynthesis of type I and III colagen during puberty]. AB - OBJECTIVES: The influence of insulin-like growth factor I (IGF-I) on the biosynthesis of type I and III collagen during puberty was studied. MATERIAL AND METHODS: Eighty-four healthy girls aged from 9.8 to 14.7 years were included into this study. After careful physical examination all girls were divided into three groups according to pubertal development. To assess pubertal stage we performed breast examination according to Tanner (bd). Group I consisted of 26 girls in prepubertal stage (1 of bd); group II (stage 2 and 3 of bd) 29 girls, and group III (stage 4 and 5 of bd)--27 girls. Serum concentration of PICP (C-terminal propeptide of type I collagen), PIIINP (N-terminal propeptide of type III collagen, as well as IGF-I were assessed by RIA methods. RESULTS: The highest mean values of serum PICP and PIIINP were found in girls in the second group (stage 2 and 3 of bd) whereas the lowest values were observed in group III (stage 4 and 5 of bd). The mean serum concentration of IGF-I presented a steady increase as puberty advanced being highest in girls in group III. No correlation was found between IGF-I and PICP and IGF-I and PIIINP. A positive correlation were observed between mean serum concentration of IGF-I and age (R = 0.52; p < 0.0001), height (R = 0.51; p < 0.0001), and body weight (R = 0.47; p < 0.0001) in investigated groups. CONCLUSIONS: Although IGF-I showed a steady increase during puberty no correlation was observed between its serum concentration and markers of collagen biosynthesis (PICP and PIIINP). PMID- 10462975 TI - [The role of galanin in the etiology of obesity in young girls]. AB - Serum galanin, FSH, LH and estradiol levels were measured in 36 young obese women and in 16 young women with normal weight and normally menstruating (control group). Obese young women were characterized by higher serum galanin levels than normally menstruating women. There were no differences in FSH, LH and estradiol levels between study and control group. These results may suggest that galanin may play a role in the pathogenesis of obesity. PMID- 10462976 TI - [The evaluation of galanin, gonadotropin and estradiol levels in girls with simple weight loss due to amenorrhea]. AB - Pathophysiology of weight loss related amenorrhea is not fully understood. Disturbances in GnRH pulsatility play a key role in this state. There is a group of neuropeptides (CRH, TRH) which inhibit GnRH secretion and also neuropeptides (galanin, Neuropeptide Y) which can stimulate GnRH release. Therefore the aim of study was to evaluate the serum galanin, FSH, LH, estradiol levels in young women with secondary amenorrhea after weight loss and in normal women Serum galanin, FSH, LH, estradiol levels were measured with the use of radioimmunoassay methods. There was no significant difference in serum galanin levels between amenorrheic women and control. FSH, LH and estradiol levels in amenorrheic women were significantly lower than in control. We conclude that serum galanin levels appear to be comparable in amenorrheic and normal women. PMID- 10462977 TI - [Analysis of liver function in patients with Turner syndrome (TS) and in patients with secondary hypogonadism with estrogen and estrogen- progestagens replacement therapy: a preliminary report]. AB - OBJECTIVES: Abnormalities of liver function tests were been reported in adults with TS and in patients with oestrogen replacement therapy. DESIGN: The aim of this study was to evaluate the prevalence of liver disease in patients before or with oestrogen therapy. MATERIALS: 79 patients, aged 13(2/12) to 60(4/12), for oestrogen replacement therapy: 13--with secondary hypogonadism (group I), 29- with TS (group II), 19--with menopause (group IV) and 18--with TS, aged 2 to 16(11/12), before oestrogen therapy (group III). METHODS: All patients were tested for liver function tests (ALAT, GGTP, total bilirubin), and patients I-III groups for serum markers of HBV and HCV infection. RESULTS: The frequency of HBV/HCV infection was 38.3% in group I, 13.7%--in group II, 16.7%--in group III. In 13.7% patients of group II and in 16.7% patients of group III the liver disease had been classified as cryptogenic. CONCLUSIONS: The prevalence of HBV/HCV infection in patients with hypogonadism and the prevalence of cryptogenic liver disease in patients with TS, seems to be relatively high. In TS liver function test should be investigated as well before oestrogen therapy and should be turned during the follow up. PMID- 10462978 TI - [Dydrogesterone in the regulation of cycle disturbances in adolescence]. AB - The aim of the study is to evaluate the dydrogesterone as an effective and safe progestin in the treatment of cycle disturbances in adolescence. The study included 116 girls, aged between 12 and 17 years, with the gynecological age 1 to 5 years. The groups included patients with painful menstruation, oligomenorrhoea and dysfunctional uterine bleeding (DUB). Different regiments of treatment with dydrogesterone were applied. Dydrogesterone in low therapeutical dose, is found to be an effective and save drug in the treatment of menstrual cycle disturbances in adolescence. PMID- 10462979 TI - [Turner syndrome in a girl with marker chromosome in karyotype]. AB - OBJECTIVES: There are suggestion, that Turner syndrome (TS) patients with mosaic karyotype for a Y-DNA-containing cell line are at risk of Y-induced gonadoblastoma. The TS patients in whom some or all cells contain a marker chromosome of unknown origin and those in whom there is clitoromegaly or other evident virillisation should be tested by FISH or PCR techniques. DESIGN: The aim of our study to present a TS girl with mosaic karyotype and marker chromosome, which origin from X chromosome was detected by FISH method. MATERIAL AND METHODS: 5-years old girl in whom TS was established. Clinical analysis included the full dysmorphic and clinical phenotype of TS. Chromosome analysis was performed on peripheral blood samples using routine cytogenetic methods and FISH technique. RESULTS: Clinical examination of girl showed many typical signs of TS besides of normal weight and length at birth and not typical for TS patients heart defect. First routine chromosome analysis, at age of 6 month, showed only 45,X cell line, Second study revealed mosaic karyotype with marker chromosome. FISH analysis for interphase nuclei and metaphase chromosomes using X centromere probe explained origin of marker from X chromosome. The karyotype was 45,X[155]/46,X,+mar[8].fish mar(X)(DXZ1+). CONCLUSION: Presence of marker chromosome in karyotype of patient with TS may modify their phenotype and it is a indication for molecular examination by FISH technique. PMID- 10462980 TI - [Age at menarche and the investigation of selected biophysical parameters in the course of menarche]. AB - DESIGN: The aim of our study was a currently clinical analysis of menarche. MATERIALS AND METHODS: The mean age of menarche was investigated in 4497 girls between 1975-1997 69 girls were examined throughly in the course of menarche. Age at menarche, body weight, height, body mass index (BMI), some of the hormones and ultrasonography of sexual organs were investigated. RESULTS: We demonstrated that mean age of menarche was 13.1 year, height 159.2 cm, weight 48 kg, body mass index (BMI) 18.9. Mean size a mucous membrane was 5.7 mm. Ratio of uterus to cervix indicated dominance of the uterus. Mean levels of hormones were: E2--89.4 pg/ml, FSH--6.2 UI/ml, LH--5.7 UI/ml. Relation LH to FSH was nearly 1. PMID- 10462981 TI - [Age at menarche in girls with celiac disease]. AB - OBJECTIVES: Delayed menarche, amenorrhea, early menopause have been sporadically reported in patients with CD. But control group matched was drawn from healthy individuals. DESIGN: The aim of this study was to investigate the age at menarche in patients with CD and in mothers of patients. MATERIALS: 59 mother/daughter pairs; daughter with coeliac disease: 49--treated with a gluten-free diet [FGD (+)] and 10 with untreated or late onset coeliac disease [FGD (-)]. METHODS: CD was diagnosed on the basis of ESPGAN criteria. All patients and mothers were asked for information on the age at menarche. RESULTS: The mean age of menarche was significantly higher in FGD (-): girls--mean 16.16 years and mother's--mean 15.49 years, than in FGD (+) patients. The mean age of menarche in FGD (+) patients and mother's were: 12.33 and 13.82 years respectively in the country (11 pairs), 13.08 and 13.49 years respectively in the little town (19 pairs), 12.90 and 13.33 years respectively in the town (19 pairs). CONCLUSIONS: The age at menarche in patients with CD and FGD is decreased to age at menarche in mother's, but is higher in the untreated CD patients and mother's. These findings support the hypothesis that the age at menarche in girls with coeliac disease is regulated by gluten-free diet and by other genetic and environmental factors. PMID- 10462982 TI - [Gonadal dysgenesis in own material]. AB - Eleven patients with Turner syndrome and 6 with pure gonadal dysgenesis were examined. Diagnosis was made on the base of clinical and cytogenetic examination. All patients had primary amenorrhea and underdevelopment of primary, secondary and tertiary sexual features. Hormonal estimations revealed elevated FSH serum concentration in women with Turner syndrome and with pure gonadal dysgenesis (46, XY) vs. to patients with 46, XX. Estradiol and Progesterone levels were low in all cases. All women were treated with estrogens and progestational agents in sequential manner with good results. We did not observe significant phenotypic differences between patients with monosomie X and patients with structural abnormalities within chromosome X and mosaicism. PMID- 10462983 TI - [Prolactinoma as a cause of primary amenorrhea in a 16-year-old girl]. AB - 16 years old girl with prolactinoma and amenorrhoea primaria was treated with Bromocorn. The symptoms of hyperprolactinemia regressed gradually--Graaf follicules started to mature in ovaries, the menstruation bleeding appeared after 12 months. The reduction of tumor mass has been obtained during this time. PMID- 10462984 TI - [Polycystic ovary syndrome in adolescent gynecology]. AB - PCO syndrome is a chronic hyperandrogenic disorder beginning in adrenarche. We tried to find a PCO share in general hyperandrogenic population of adolescent girls regarding hormonal and usg parameters. We found LH v FSH ratio > 2:1 as sensitive feature of PCO for most of the patients then other clinical and endocrinological criteria. PMID- 10462985 TI - [Acantosis nigricans in polycystic ovary syndrome]. AB - Acantosis Nigricans (AN) is a skin disease with rough or velvet areas present on neck or as hyperpigmented and wrinkled skin. AN often forms HAIR-AN syndrome consisted of hyperandrogenizm, insulin resistance and AN. We present AN and PCO syndrome in 13 yrs old girl treated by laparoscopic wedge resection. We concluded that laparoscopic procedure is a diagnostic measure and mode of PCO treatment. Decreased hyperandrogenizm resulted in AN appearance withdrawall. PMID- 10462986 TI - [Repeated amenorrhea in an adolescent girl in the course of flood disaster in Klodzko Region, July 1997]. AB - OBJECTIVES: A natural disaster has been defined as a disruption of human ecology that exceeds the capacity of the community to function normally. DESIGN: After the flood disaster in Klodzko Region, July 1997, the major problem in female adolescents was observed: secondary amenorrhea. MATERIALS AND METHODS: 17 female adolescents, aged 13-18, which injured from the flood disaster with secondary amenorrhea were investigated. A control random group consists of 17 girls diagnosed before oral contraception. Diagnostic work-up includes history, physical and psychological examinations, hormonal profiles, transvaginal ultrasonography, color Doppler analysis of utero-ovarian arterial blood flow. RESULTS: FSH, LH, E2 plasma levels and LH/FSH ratio were significantly lower in the amenorrheic group compared to normal girls. Prolactin serum levels after metoclopramid administration were significantly higher in the amenorrheic group. Lower impedance to blood flow in the intraovarian arteries have been shown. Psychosomatic disorders related to hypothalamic amenorrhoea were diagnosed. CONCLUSIONS: Psychosocial stress observed during the flood disaster caused hypogonadotropic hypogonadism amenorrhoea in the female adolescents. PMID- 10462987 TI - [Repeated amenorrhea after body weight loss. Efficacy of treatment of estrogen and gestagen depending on the body mass index]. AB - The aim of the study was to evaluate end effect of different doses of estradiol and BMI changes by the time of therapy and BMI changes before therapy. We found that most important is to increase body mass to have regular cycles and there is no difference from statistical point of view to use different doses of estradiol (constant v. step down protocol). We used different progestogenic agents: medroksyprogesteron and dydrogesteron. PMID- 10462988 TI - [Primary amenorrhea in patient with persistent communicating hydrocephalus]. AB - We report a case of primary amenorrhea caused by persistent communicating hydrocephalus. Patient referred to our outpatient clinic because of primary amenorrhea. With the use of computer tomography diagnosis of hydrocephalus was established. After treatment: implantation of ventriculo-peritoneal valvue spontaneous regular menses occurred. PMID- 10462989 TI - [Course of pregnancy and delivery in adolescence]. AB - An increasing number of deliveries in adolescents is a growing problem, not only obstetrical but also social in the whole world. Pregnancy is a physiological condition, although with girls under 18 needs a special care and treatment and is called as a high risk pregnancy. Especially girls with age under 16 appear to have high risk of pregnancy pathology as well as low-birthweight infants. PMID- 10462990 TI - [The premenstrual syndrome frequency and the intensity of its symptoms in young women and teenagers living in the Upper Silesian Industrial region]. AB - Contemporary home and world medical literatures deal at length with somatic and psychopathologic symptoms connected with changes taking place during the menstrual cycle. The symptoms bring about changes in women's behaviour, life activities and interpersonal relations. The majority of women suffer during the premenstrual period from mood disorder and somatic pains. Nevertheless, the presence of the pains does not diagnose PMS. Over many years PMS has not been clearly defined. Nowadays, making use of precisely-defined criteria, we are able to specify the number of women suffering from the syndrome. Applying the criteria compiled by American Psychiatric Association, we analyse the frequency of PMS occurrence. On the basis of the criteria 180 young women ages 17-25 living in the Upper Silesia Industrial Region have been interviewed. Besides PMS clearly defined diagnostic criteria the survey included also questions about how the intensity of disorders related to the course of menstrual cycle influences the women's feelings of well-being. The questionnaire scores demonstrate that only few women suffer from PMS defined by American Psychiatric Association, but its symptoms have considerable effects on the women's lives. Simultaneously, many women suffer from numerous pains and disorders which have not been diagnosed as PMS, but which should be examined separately. PMID- 10462991 TI - [The assessment of selected markers of bone tissue metabolism and bone mineral density in girls with resulting from forced weight reduction]. AB - The study comprised the assessment of the effects of secondary amenorrhea on bone tissue metabolism and bone mineral density in girls after forced weight reduction. The study concentrated on assessing and analysing the levels of selected markers of bone tissue remodelling and bone mineral density. PMID- 10462992 TI - [Assessment of the bone mass density in young women with secondary amenorrhea]. AB - Analisys of results of bone mass density among 120 patients of Gynecological Endocrinology Clinic AM Poznan was presented. Endocrinological background of lowered bone mass density was viewed. PMID- 10462993 TI - [The evaluation of blood flow in uterine arteries in girls with polycystic ovary syndrome by transvaginal color Doppler ultrasonography]. AB - OBJECTIVE: The aim of this study was to evaluate of blood flow in uterine arteries in girls with PCOS and to determine whether uterine blood flow variations are associated with endocrine findings typical of PCOS. METHODS: Seventy two girls with clinical, endocrinologic and ultrasonographic features of PCOS (mean age 18.8 +/- 1.3 years) were examined. The control subject consisted of twenty girls with regular, ovulation cycles. None of these girls had received hormonal treatment for at least 6 months before the study. Pelvic ultrasound examination was made in each patient with Acuson 128 XP/10 with the 4/5/7 MHz transvaginal transducer. Examination was performed in early follicular phase. Blood flow was made in each patient in the ascending branches of uterine arteries. Index values pulsatility index (PI) and resistance index (RI) were used for the purpose of better comparison of the data. Associations between values of PI and RI and hormones (testosterone, luteinizing hormone) were examined. RESULTS: No significant differences between PI and RI of the left and right uterine artery was observed. PI in PCOS girls was mean 3.19 +/- 0.52 ranged from 2.1 to 4.3, in control girls PI was mean 2.33 +/- 0.36 ranged from 1.6 to 3.1. The PI was positively correlated (r = 0.435, p < 0.01) with testosterone and PI was positively correlated (r = 0.450, p < 0.001) with LH. The RI in PCOS patients was mean 0.87 +/- 0.04 ranged from 0.74 to 0.93, in control girls 0.72-0.87, mean 0.80 +/- 0.04. The RI was positively correlated (r = 0.466, p < 0.001) with testosterone and RI was positively correlated (r = 0.492, p < 0.001) with LH. CONCLUSIONS: The elevated uterine artery resistance was observed in girls with PCOS. PMID- 10462994 TI - Nosocomial gram-negative bacteremia in critically ill patients: epidemiologic characteristics and prognostic factors in 147 episodes. AB - Although gram-positive organisms are the most common causes of nosocomial bloodstream infections, gram-negative bacteremia carries higher risks of severe sepsis, septic shock, and death among critically ill patients in intensive care units (ICUs). We performed a prospective epidemiologic analysis of nosocomial gram-negative bacteremia episodes among ICU patients and sought to identify risk factors for mortality among these patients. All episodes of nosocomial gram negative bacteremia documented in five ICU wards of our hospital during a 2-year period were included. There were 147 episodes (124 patients) of gram-negative bacteremia documented during the study period. The overall mortality rate was 36.1%, and 77.4% of all deaths were directly related to the bloodstream infection. Gram-negative bacteremia was associated with prolonged ICU stay (45.7 d vs 6.1 d for all ICU patients). The most common isolate was Acinetobacter baumannii, followed by Burkholderia cepacia and Enterobacter cloacae. The most frequent source of infection was the lower respiratory tract (32.0%). Of the agents tested, ciprofloxacin, imipenem, and ceftazidime were the most active against the clinical isolates. Multivariate logistic regression analysis identified the presence of septic shock (odds ratio, OR = 17.66, p < 0.001) and rapidly fatal and ultimately fatal underlying conditions (OR = 3.47, p = 0.032) as being independent risk factors for mortality. Early appropriate antibiotic treatment did not result in significant improvement in survival. These findings suggest that prevention of lower respiratory tract colonization and nosocomial pneumonia are crucial for reducing the incidence of nosocomial gram-negative bacteremia in the ICU. Serious underlying illnesses and septic shock were the most important risk factors for death in these patients. PMID- 10462995 TI - Prognostic determinants of infective endocarditis in the 1990s. AB - The clinical profiles of infective endocarditis (IE) might have changed in recent years, owing to the advent of transesophageal echocardiography, the introduction of new diagnostic criteria, and the increased frequency of intravenous drug abuse. In this retrospective study, we sought to identify factors affecting the in-hospital outcome of IE patients in a single tertiary referral center in the 1990s (1990-1997). Eighty-eight episodes of IE in 80 consecutive patients admitted from January 1990 through June 1997 were evaluated. Clinical variables that were significantly associated with in-hospital mortality in univariate analyses were entered into a multiple logistic regression model. A total of 22 patients (25%) died. Fatal episodes were significantly more likely than non-fatal episodes to involve older patients (> or = 50 years), use of coumadin, short interval between symptom onset and hospitalization (< 15 days), noncardiac shock, and complications of the heart, central nervous system, and kidneys. White blood cell counts and C-reactive protein concentrations were also significantly higher in fatal than in non-fatal episodes of IE. Multivariate analysis showed that in hospital mortality was associated with noncardiac shock, neurological complications, cardiac complications, and older age (> or = 50 years). Compared with previous reports, our findings suggest that the clinical profiles of IE have undergone some changes in the 1990s. The most important prognostic predictors of in-hospital mortality in patients with IE were noncardiac shock and neurologic complications. PMID- 10462996 TI - Effects of dipyridamole, nitroglycerin, and nitroprusside on coronary vascular resistance in rabbits: measurement with pulsed Doppler velocimetry. AB - Dipyridamole, nitroglycerin, and nitroprusside are all effective vasodilators. However, few studies have compared their in vivo coronary vasodilatation effects. The purpose of this study was to compare the effects of these agents on coronary blood flow velocity and vascular resistance in anesthetized, open-chest rabbits. Male New Zealand white rabbits were anesthetized, and a 3-mm, suction-type, pulsed Doppler velocimeter probe was applied to the proximal part of the left anterior descending coronary artery after median sternotomy. The rabbits received infusion of various doses of dipyridamole (0.1 mg/kg, n = 5; 1 mg/kg, n = 5; 10 mg/kg, n = 9), nitroglycerin (0.01 mg/kg, n = 8; 0.1 mg/kg, n = 5; 1 mg/kg, n = 5; 10 mg/kg, n = 7), or nitroprusside (0.01 mg/kg, n = 5; 0.1 mg/kg, n = 5; 1 mg/kg, n = 5). The percent changes in coronary blood flow velocity and coronary vascular resistance were measured. All three vasodilators increased coronary blood flow velocity significantly and decreased coronary vascular resistance dose dependently. The dose-response curves of dipyridamole, nitroglycerin, and nitroprusside were significantly different from one another (p < 0.01). Dipyridamole at a dose of 10 mg/kg produced the greatest increase in coronary blood flow velocity and the greatest reduction in coronary vascular resistance in anesthetized, open-chest rabbits. PMID- 10462997 TI - Comparison of indoor allergens, allergic scores, and demographic data in Taiwanese adults with asthma or allergic rhinitis, or both. AB - We examined the relationships between exposure to different indoor allergens and demographic factors in 515 adult Taiwanese patients with allergic diseases such as asthma and rhinitis. Patients who had positive screening results on an immunoglobulin E (IgE) test were recruited from the outpatient clinics of Chang Gung Memorial Hospital. Patients were divided into the following three groups: asthma (n = 332), combined asthma and allergic rhinitis (n = 165), and allergic rhinitis (n = 18). Serum samples were analyzed for IgE by enzyme-linked immunosorbent assay. Significantly elevated titer of dust mite-specific IgE were found in all three groups, with markedly higher levels in the combined asthma and allergic rhinitis group (p < 0.05). Allergen scores were significantly related to the demographic variables of older age (> or = 65 years), female gender, and spring season, with significantly lower scores (p < 0.05). No significant difference in allergen scores was found among cigarette smoking subjects and subjects from residential areas. Multiple linear regression analysis revealed significant allergic contributors to be, in order of importance, age, gender, and atopic disease category. We conclude that mite allergen exposure is the most significant factor associated with asthma and/or allergic rhinitis in Taiwan. Younger subjects and males had a higher rate of asthma and/or allergic rhinitis. PMID- 10462998 TI - Association of allergy with Tourette's syndrome. AB - In our clinical practice, we often encounter signs and symptoms of allergy, such as rhinitis and asthma, in patients with Tourette's syndrome (TS). Some of the allergic manifestations are similar to the oral tics or motor tics found in TS patients. To clarify the association between TS and allergy, we evaluated 72 consecutive patients with TS from 1 September 1996 through 31 August 1997. The diagnosis of TS was based on the Diagnostic and Statistical Manual of Mental Disorders diagnostic criteria. Sixty-five boys and 7 girls, 4 to 17 years old (9.4 +/- 3.1 yr) were evaluated using the Multiple Allergens Simultaneous Tests (MAST) for the detection of total and specific immunoglobulin. Forty-five patients had positive results, of whom 41 (56.9%) had clinical evidence of allergy. The prevalence of allergy in the local population as reported by The International Study of Asthma and Allergy in Childhood Taiwan Group (1994) was 44.3% (33.5% with allergic rhinitis and 10.8% with asthma). These subjects served as controls. Comparing the number of patients with clinical evidence of allergy in the MAST positive group (56.9%) of TS patients with the control group (44.3%), the difference was significant++ (p < 0.05). The prevalence of allergy in TS patients in our study was significantly higher than in the general population. TS had an association with allergy. PMID- 10462999 TI - Annual risk of tuberculous infection in Taiwan, 1996-1998. AB - Tuberculosis is still an important public health issue in Taiwan, and monitoring the trend of annual risk of infection (ARI) with Mycobacterium tuberculosis is essential. In this study, we conducted tuberculin skin tests to estimate the prevalence and annual risk of M. tuberculosis infection in first-grade schoolchildren in Taiwan Province. Because mass bacille Calmette-Guerin (BCG) vaccination programs have been carried out here, only non-BCG-vaccinated students were tested. From September 1996 through June 1998, there were 520,866 registered first-grade elementary school students in Taiwan Province. Of them, 15,147 (2.9%) were non-BCG-vaccinated, as determined by the absence of a BCG scar. All of them were tested for M. tuberculosis infection with 1 tuberculin unit (0.1 mL injection) of purified protein derivative RT23, by means of the Mantoux technique. Among the tested schoolchildren, 430 (2.8%) had a positive tuberculin reaction. Thus, the calculated ARI was 0.44%. The ARI varied in different areas of Taiwan, being highest (1.04%) in Nantou County and lowest (0.14%) in Miaoli and Tainan Counties. The ARI in aboriginal areas (1.16%) was 2.7 times that in nonaboriginal areas (0.42%). Our results indicate that the M. tuberculosis ARI is still high in Taiwan. To achieve the World Health Organization target of less than 0.1% for industrialized countries, we must intensify tuberculosis control programs in Taiwan. PMID- 10463000 TI - Comparison of fluid-based, thin-layer processing and conventional Papanicolaou methods for uterine cervical cytology. AB - This study was conducted to compare the diagnoses and specimen adequacy of the ThinPrep (Cytyc Corporation, Boxborough, MA, USA) and conventional Papanicolaou preparation methods for cervical cytology. Cervical specimens from 972 patients from our gynecology clinic were analyzed. A single sample of the uterine cervix taken from each patient was first used to prepare the conventional Papanicolaou smear. Afterward, the residual tissue on the sampling device was rinsed into a fluid preservative from which two slides were prepared by means of the ThinPrep method. Conventional and ThinPrep slides were examined independently. Cytologic diagnoses and specimen adequacy were classified according to the Bethesda System. The diagnoses based on the ThinPrep and conventional smears matched in 939 (96.6%) cases. Low-grade squamous epithelial or higher grade lesions were detected with at least one of the methods in 59 cases and with both methods in 42 cases. Fifteen more cases of low-grade squamous epithelial or higher grade lesions (58 vs 43; p < 0.001) and 10 more cases of high-grade squamous epithelial lesions or carcinoma (42 vs 32; p = 0.006), were detected with ThinPrep than with conventional smears. The ThinPrep method yielded a higher rate of specimens that were satisfactory for cytologic examination than the conventional smears (89.8% vs 70.9%, p < 0.001). The detection rate of infectious agents was also higher with the ThinPrep method (14.9% vs 6.6%; p < 0.001). The reproducibility rate of the twin slides prepared with the ThinPrep method was greater than 99%. In conclusion, the ThinPrep method yielded a significantly higher positive detection rate of cervical abnormalities and more satisfactory specimens than conventional Papanicolaou smears. PMID- 10463001 TI - Combination of color Doppler ultrasonography and ultrasound-guided fine-needle aspiration cytology for localization of parathyroid lesions. AB - We examined the usefulness of color Doppler ultrasonography combined with ultrasound-guided fine-needle aspiration cytology (FNAC) for preoperative localization of parathyroid lesions in patients with hyperparathyroidism. Twenty eight patients (19 men, 9 women; mean age, 56 years) with clinical and laboratory evidence of hyperparathyroidism underwent preoperative color Doppler ultrasonography and ultrasound-guided FNAC prior to surgery. On ultrasonography, the parathyroid lesions were anechoic or hypoechoic, located behind or at the margin of the thyroid gland, and mimicked blood vessels in some cases. On color Doppler ultrasound, parathyroid lesions showed little or no vascularity. They were easily differentiated from blood vessels, thus improving the safety of ultrasound-guided FNAC. Thirty-four (76%) of 45 surgically confirmed parathyroid lesions were detected with ultrasonography in 21 (75%) of the patients. There was no significant difference in the detection rates of lesions in the right upper, right lower, left upper, and left lower parathyroid glands. Eight (29%) patients had coexisting thyroid lesions. FNAC of the ultrasound-detected abnormalities provided preoperative confirmation that they were parathyroid lesions. In conclusion, our findings show that color Doppler ultrasonogrphy in combination with ultrasound-guided FNAC is useful for preoperative localization of parathyroid lesions. PMID- 10463002 TI - Intellectual outcomes of patients with congenital hypothyroidism not detected by neonatal screening. AB - Mental retardation is a major sequela of delayed treatment of congenital hypothyroidism. In this study, we investigated the intellectual outcomes of patients with congenital hypothyroidism diagnosed before a nationwide screening project started. Eighty-two patients had intelligence tests done while in the euthyroid state. Their mean intelligence quotient (IQ) was 78 +/- 21 (+/- SD) (n = 79). The mean verbal IQ was significantly higher than the mean full-scale IQ. The mean IQs of both ectopic thyroid (84 +/- 21, n = 28) and dyshormonogenic patients (85 +/- 14, n = 18) were significantly higher than that of patients with athyrosis (64 +/- 18, n = 19) (p < 0.05). Patients who were treated before 3 months of age had significantly higher mean IQs (90 +/- 16, n = 16) than those treated after 3 months (75 +/- 21, n = 63) (p < 0.01). Our results confirm that both patients with ectopic thyroids and those with dyshormonogenesis have better intellectual outcomes than athyrotic patients do, and that early treatment improves the intellectual outcome of patients with congenital hypothyroidism. PMID- 10463003 TI - Remitting seronegative symmetrical synovitis with pitting edema (RS3PE syndrome). AB - A 63-year-old man presented with acute symmetrical polysynovitis associated with pitting edema of both the hands and feet. He was seronegative for rheumatoid factor and no radiologically evident erosion was noted in the joints of his hands and feet. Evaluation excluded congestive heart failure, nephrotic syndrome, and hypothyroidism as the cause of edema. Treatment with nonsteroidal anti inflammatory drugs and low-dose steroids induced complete remission. The clinical manifestations of this patients were consistent with those of a distinctive, although rare, form of arthritis called remitting seronegative symmetrical synovitis with pitting edema (RS3PE) syndrome. This syndrome has a good prognosis in elderly patients. PMID- 10463004 TI - Varicella arthritis diagnosed by polymerase chain reaction. AB - We report a 2-year-old girl who developed acute arthritis of the left knee 4 days after the onset of a typical varicella infection. She was first thought to have pyogenic arthritis caused by Staphylococcus aureus. Accordingly, oxacillin was administered upon hospitalization. On the third day after hospitalization, bacterial cultures of the synovial fluid and blood showed no growth and oxacillin was discontinued. Although a viral culture of the synovial fluid for varicella zoster virus (VZV) was negative, varicella DNA was identified by means of polymerase chain reaction (PCR) with VZV-specific primers. The patient recovered spontaneously. To differentiate this condition from septic arthritis is important. PCR is a sensitive technique that can demonstrate the presence of VZV DNA in synovial fluid, even if viral cultures are negative. PMID- 10463005 TI - Adult non-Hodgkin's lymphoma. PMID- 10463006 TI - Why physicians and lay people smoke and how can it be reduced? AB - OBJECTIVE: The objective of this study was to find out the level of knowledge the physicians and lay people have pertaining to the effect of cigarettes, why certain physicians smoke and what measures could be applied to reduce the rate of smoking. METHODS: A questionnaire was administered to the one hundred physicians who smoke, one hundred non-smoking physicians and one hundred lay people who smoke to determine their attitude towards this addition. Subjects were chosen using convenience sampling. The physicians were picked from six hospitals of Karachi. RESULTS: When the smoking physicians were asked what could motivate them to stop smoking, majority of them said that an occurrence of a smoking related illness would. Majority of the physicians who do not smoke felt that individual will was the greatest force keeping them from smoking. When asked how smoking can be reduced in Pakistan, majority of the physicians, both smoking and non-smoking, favoured mass health education. Lay smokers expressed marked ignorance about deleterious effects of cigarette smoke. Like smoking physicians, majority of them said that occurrence of an illness related to smoking would effectively motivate them to stop smoking. CONCLUSION: Based on this survey we conclude that mass health education and enforcement of the ban on smoking in public places will effectively reduce the number of smokers. There is a need to educate physicians and the general public about the cardiac and carcinogenic effects of smoking. PMID- 10463007 TI - Patients view of the anaesthetist in a developing country. AB - OBJECTIVE: The objective of this study was to assess patients regarding their knowledge and perception of the anaesthetists' role in patient care in a developing country. SETTING: The audit was conducted at the Aga Khan University Hospital, Karachi. METHODS: Two hundred surgical and 102 medical out-patients were included in the audit prospectively who filled a detailed questionnaire. RESULTS: Fifty-six percent of the patients knew that anaesthetists were medically qualified but only 20% and 27% had an idea about their role in the intraoperative and immediate postoperative period. 14.6% were aware of the anaesthetists role outside the operating room. No difference was observed between the two groups of patients except that a higher percentage of surgical patients were interested in having more information regarding anaesthesia and wanted to choose their anaesthetist if the need arose. Gender difference was observed in answer to two questions only. A higher percentage of males know the role of anaesthetist in the recovery room whereas a greater number of females who had surgery before remembered their anaesthetist postoperatively. CONCLUSION: The results of this audit show a poor perception of the anaesthetists role both inside and outside the operating room. Although public knowledge surveys have been carried out in developed countries there is a pressing need to repeat these in developing countries since the image as perceived by public may have a direct bearing on the recruitment of young doctors into an already shortage specialty in these less affluent countries. PMID- 10463008 TI - Prevalence of macrovascular complications in diabetics of WAH, District Rawalpindi. AB - OBJECTIVE: To determine the prevalence of macrovascular complications in diabetic subjects with relation to age duration of diabetes and metabolic control. SETTING: Primary care centres of WAH, District Rawalpindi cross sectional population based study. METHODS: A total of 805 known diabetics (380 males and 425 females) were studied. Relevant history, detailed physician examination and ECG were done to assess the presence of ischaemic heart disease, stroke and diabetic foot complications. Their diabetic control was assessed by estimation of their plasma glucose and glycosylated haemoglobin. RESULTS: The prevalence of macrovascular complications was as follows: Ischaemic heart disease 19.8% (female 4.7%, males 7.8%), cerebrovascular disease 6.2% (females 4.7%, males 22.1%) and diabetic foot complications 2.1%. The macrovascular complications increased with age, duration of diabetes and were more prevalent in uncontrolled diabetics having glycosylated haemoglobin > 8.6%. CONCLUSION: Prevalence of macrovascular complication in our population is higher as compared to the West probably due to better management and diabetic care in those countries. PMID- 10463009 TI - Non-Hodgkin's lymphoma in Pakistan: a clinicopathological profile of 175 patients. AB - OBJECTIVE: Non-Hodgkin's Lymphomas (NHL) are common in Southeast Asia, Middle East and Africa. The study was primarily undertaken to establish the demographic and clinicopathologic features of NHL in Pakistan. A secondary aim was to determine the applicability of the International Prognostic Index (IPI) developed by the international lymphoma task force in a developing country. METHODS: A retrospective and prospective analysis was performed on 175 patients over 14 years age who presented to the Department of Oncology between August 1994 and December 1996. All patients underwent initial staging according to Ann Arbor Staging System. Histopathologic classification was done according to the International Working Formulation. The IPI was applied to patients with aggressive lymphomas and age adjusted index to patients < 60 years, complete remission (CR), disease free survival (DFS) and overall survival (OS) were calculated. RESULTS: All 175 patients were evaluable. Median age of our patients was 45 years. Male to female ratio was 1.9:1. Seventeen (9.7%) patients were classified as low grade lymphomas while 158 (91.3%) had intermediate and high grade NHL. Large cell lymphoma was present in 30.9% patients. CHOP (cyclophosphamide 650 mg/m2 day 1, vincristine 1.4 mg/m2 day 1, doxorubicin--45 mg/m2 day 1 and prednisone 100 mg/m2 day 1-5) was the most common chemotherapy regimen used. Advanced stage (74.9%) > B symptoms (51%) and extranodal involvement (74.3%) were present. One hundred sixty-seven patients were evaluable for response of which 42.8% achieved CR. Median DFS was 19 months and median OS was 22 months. The IPI was applicable to 153 patients and age adjusted IPI to 124 of 153 patients. CONCLUSION: Aggressive histology, extranodal diseases, B symptoms and advanced disease are common. International index and age adjusted international index predicted outcome accurately in various risk groups. PMID- 10463010 TI - Rapid diagnosis of malaria--a new approach. PMID- 10463011 TI - Salmonella typhi meningitis. PMID- 10463012 TI - View box--case I. Pulmonary alveolar proteinosis. PMID- 10463013 TI - Home-care of the dying. PMID- 10463014 TI - Effect of prepregnancy body mass index and gestational weight gain on birth weight. AB - OBJECTIVE: Nutritional status of women has been considered an important prognostic indicator of birth outcome. The study aims to show the effect of various prepregnancy Body Mass Index (BMI) categories and corresponding gestational weight gain on newborn birth weight. METHODS: Two hundred women were included in the study. These women had regular antenatal visits and later delivered at The Aga Khan University Hospital (AKUH) between the period January 1, 1996 to December 31, 1997. RESULTS: For women with prepregnancy BMI < 19., mean birth weight of newborns was lower for those gaining < 12.5 kg than those gaining > 12.5 kg (P < 0.001). Women who started their pregnancy with BMI 19.8-26 and gained weight above expected range gave birth to high birth weight babies (P = 0.009). Gestational weight gain did not have a significant association with birth weight for women having prepregnancy BMI > 26. CONCLUSION: Efforts should be made to attain adequate prepregnancy weight to reduce the likelihood of low birth weight babies. Hence, special attention should be paid to women with low prepregnancy BMI. PMID- 10463015 TI - Gait analysis in the therapeutic environment. AB - Gait analysis is one aspect of the overall assessment of any patient with a movement disorder. Loss of walking ability is often a major issue to the patient, thus justifying the length of time that should be spent in establishing gait problems and planning re-education. Biomechanical measures of kinematics, kinetics and electromyographical activity are essential to give a complete picture of the specific gait characteristics (Perry 1992; Whittle 1996b). However, observational analysis remains the most commonly used tool in the therapeutic situation (Patla et al. 1987). Reliability of observational analysis is poor (Krebs et al. 1985) and therapists should be encouraged to use objective measures to give a more representative account of the gait pattern. A systematic approach to data collection and recording should be adopted and the key kinematic data of walking including velocity, stride length, base of support and joint angles, should be collected in order to provide a baseline on which to measure clinical effectiveness. Valid outcome measures must be established to evaluate gait effectively to afford therapists the ability to assess their treatment of gait deviations. An understanding of biomechanical terminology is essential to aid the selection of appropriate gait analysis tools and for interpretation of the results. PMID- 10463016 TI - Imaging of the painful shoulder. AB - A variety of imaging techniques are available for evaluating shoulder pathology. The common disorders of impingement, rotator cuff tears, biceps tenosynovitis and instability are discussed along with the role of various imaging modalities in establishing their diagnosis. Plain radiographs can provide useful information particularly with a history of trauma but give limited information on the soft tissue structures around the shoulder. Ultrasound is a useful and inexpensive means of assessing the biceps tendon and rotator cuff but has a number of limitations and varying reports on its accuracy. Computed tomography (CT) is most helpful in the evaluation of shoulder trauma but gives limited information on the soft tissues. When performed with intra-articular contrast, CT can be used to evaluate shoulder instability particularly in cases where magnetic resonance imaging (MRI) is not available or contraindicated. MRI with or without intra articular contrast is the most accurate imaging modality for evaluating shoulder pathology allowing visualization of the soft tissues that are often the source of pain. PMID- 10463017 TI - Is there a positional fault at the inferior tibiofibular joint in patients with acute or chronic ankle sprains compared to normals? AB - The purpose of this study was to test the hypothesis that a positional fault occurs at the distal tibiofibular joint when the foot is inverted beyond its normal range (Mulligan 1995). Seventeen normal subjects, two subjects with a chronic tendency to 'go over' on one or both ankles and six patients with acutely sprained ankles were positioned in supine lying with the foot to be tested in the subtalar neutral position. An anteroposterior (AP) mobilization was then performed on the lateral malleolus of the right and left ankles of all those included in the study. The movement that occurred at the distal fibula as a result of this mobilization was recorded and plotted against the force required to produce it. The slopes of the graphs from the three groups of subjects were statistically analysed using the appropriate t tests. Results indicated that a significantly greater amount of movement per unit force occurred in one third of the patients with acutely sprained ankles (P = 0.01%, P = 0.09%). These results lend support to the hypothesis that a positional fault occurs at the inferior tibiofibular joint in ankle sprain patients. This could have exciting implications for the future direction of the treatment of these injuries. PMID- 10463018 TI - Use of lumbar extension, slump test, physical and neurological examination in the evaluation of patients with suspected herniated nucleus pulposus. A prospective clinical study. AB - This prospective and consecutive study was designed to evaluate the validity of different clinical tests, e.g. lumbar extension in lying and slump test for patients with suspected herniated nucleus pulposus, in comparison with findings on computed tomography (CT) and/or magnetic resonance imaging (MRI) scan. There were 105 patients who were seen and examined by the senior author (for the sake of the study) at the Orthopaedic Physiotherapy Department, on an average of 5.5 days (range 0-21 days) before CT and/or MRI examination were carried out. There were 36 women and 69 men with an average age of 42.7 +/- 9.8 (range 19-64) years. According to the radiological findings on CT and/or MRI, the patients were divided into three groups: 52 patients with disc hernia, 41 patients with bulging discs and 12 patients without positive findings. The mean values with standard deviations of 25 variables of three diagnostic groups were studied. Multiple comparison adjustment according to Bonferroni showed significant differences for three variables that were of diagnostic value (lumbar range of motion for forward flexion, left side-bending in standing, and pain distribution during extension in standing). The agreement between clinical and radiological findings for type and level of diagnosis of disc herniation was accurate in 72 patients (69%). The diagnostic sensitivity for disc herniation was 82.6% and the specificity 54.7%. PMID- 10463019 TI - A qualitative assessment of shoulder girdle elevation during the upper limb tension test 1. AB - The 'feel through range' and the 'end-feel' of the elbow extension, and the elevation of the shoulder girdle during the upper limb tension test for the median nerve are considered important in clinical decision making. As no objective measurements have been published regarding these parameters, a special device was designed in order to obtain objective information on the force by which the shoulder girdle is elevated during the test. The purpose of this technical note is to describe the technical aspects of the device and its practical application. In a pilot study, five asymptomatic subjects have been assessed. The relationship between the elevation of the shoulder girdle and the range of elbow extension has been analysed. The data of five subjects and representative curves of one subject are presented. All five subjects demonstrate a considerable increase in force elevating the shoulder girdle. The results show that the device can be used to measure the force by which the shoulder girdle is elevated throughout the test. Although the number of subjects and the number of repetitions is limited, the data suggest that the method is accurate and that the phenomenon of shoulder girdle elevation during the upper limb tension test in asymptomatic subjects is consistent. PMID- 10463020 TI - Perspectives and relevance of medico-legal work. PMID- 10463021 TI - Is the pain and dysfunction of shoulder impingement lesion really second rib syndrome in disguise? Two case reports. PMID- 10463022 TI - Longitudinal study of recruitment and retention in small community homes supporting persons with developmental disabilities. AB - Variables associated with turnover in 110 small Minnesota group homes supporting people with developmental disabilities between December 1993 and December 1996 were examined. Common staffing problems included recruiting qualified workers (reported by 57% of supervisors) and staff turnover (44%). Turnover for direct support professionals was 46%. Of workers who left, 45% left within 6 months of hire, and another 23%, between 6 and 12 months. A multiple regression analysis (with 11 variables) accounted for 34% of the variability in facility turnover rates. Variables contributing significantly to the prediction of higher turnover included greater support needs of residents, lower starting pay, less tenured supervisors, and fewer direct support professionals eligible for paid leave. Implications for practice, research, and policy were discussed. PMID- 10463023 TI - Cross-validated factor structures and factor-based subscales for PASS and PASSING. AB - Program Analysis of Service Systems (PASS 3) and Program Analysis of Service Systems' Implementation of Normalization Goals (PASSING) are used internationally to assess service quality in mental retardation and other fields. In two studies, we derived and cross-validated the factor structures of these program evaluation instruments. In Study 1, based on 626 PASS 3 evaluations, four factors provided a good fit to the cross-validation data. In Study 2, based on 633 PASSING evaluations, three factors furnished a good fit to the cross-validation data. Similarities between the two factor structures, parallels with the structural and functional service dimensions emphasized in ecological approaches to evaluation, and practical uses in program evaluation of subscales based on the factors were discussed. PMID- 10463024 TI - Treatment of self-injurious behavior and quality of life for persons with mental retardation. AB - Self-injurious behavior (SIB) is a highly problematic and damaging behavior with profound implications for a person's quality of life. Despite numerous reports documenting changes in self-injury, it is not well-known how these changes relate to systematic improvements in quality of life. We surveyed 41 journals from 1978 to 1996 to identify use of quality of life outcome measures following self-injury treatment. A sample of 138 research articles involving 436 subjects with self injury was reviewed. Forty articles (29%) were found that contained some quality of life measure. Very little detailed information was available concerning the specific nature of change in life-style based on immediate changes in SIB. This lack of information and its possible implications are discussed. PMID- 10463025 TI - Profile of advisors to self-advocacy groups for people with mental retardation. AB - An extensive profile of self-advocacy group advisors was developed. A telephone survey was administered to 118 advisors randomly selected from the Longhurst (1994) Directory of Self-Advocacy Groups; 90 surveys were completed (76%). Major findings are reported in the areas of advisor demographic data, employment and advising experience, current employment profile, and recruitment. Results provide important baseline information for individuals affiliated with the self-advocacy movement, UAPs, developmental disability councils, Arc chapters, and professional organizations. These results can be used to help better understand the people who are currently supporting self-advocacy groups and to identify possibly support person recruitment issues. PMID- 10463026 TI - Validity of the Stanford-Binet Intelligence Scale-IV: its use in young adults with mental retardation. AB - The validity of the Stanford Binet-IV (SB-IV) was assessed. This test and the WAIS-R and WRAT-R were administered to 42 adults previously classified with mild to moderate mental retardation. Validity coefficients between scores on the SB-IV and the other two measures were significant. The mean IQ on the SB-IV (mean Test Composite = 43.26) was significantly lower than that on the Wechsler Adult Intelligence Scale-Revised--WAIS-R (mean Full-Scale IQ = 57.91). With regard to the internal validity of the SB-IV, the intersubtest relationships of each of the four Area scores correlated significantly with the Test Composite (range = .66 to .91). Verbal Reasoning earned the highest correlation (.91). Results support the SB-IV's concurrent, criterion-related, and internal validity for use with young adults who have mental retardation. PMID- 10463027 TI - Commentaries on the presidential address of Robert Schalock. An odyssey down the yellow brick road. PMID- 10463028 TI - Commentaries on the presidential address of Robert Schalock. More stories behind the stories? PMID- 10463029 TI - Impact of race, poverty, and ethnicity on services for persons with mental disabilities: call for cultural competence. PMID- 10463030 TI - Medicaid HCBS "Waiver" recipients are now twice the number of Medicaid ICF/MR residents. PMID- 10463031 TI - [Pathophysiology of Helicobacter pylori infection]. AB - The (re)discovery of the gastric pathogen Helicobacter pylori almost one and a half decade ago completely changed our conception on gastroduodenal ulcer disease and gastric cancer. The most important issue is that Helicobacter pylori induces a mild, antrum dominant chronic pangastritis without increased risk of severe diseases in most of the cases. A smaller part of the infected develops antrum dominant gastritis with increased risk of duodenal ulcer. In a few cases the corpus is also affected, pangastritis occurs with increased risk to develop gastric ulcer and gastric cancer. Until now it is not quite obvious why some of the infected patients get ill while others not, and why different diseases develop in different patients. 1. From the bacterial point of view it is ascertained that toxin-producing strains (VacA, CagA positive) are more likely to induce ulcer formation and gastric carcinoma. 2. Genetic phenotype of the infected patients (blood-group and HLA antigens) may also be of importance. 3. Environmental factors may affect (promote or inhibit) disease development. All of these factors determine the complex immunological, functional and morphological changes characteristic for the developing disease. PMID- 10463032 TI - [Long-term results with fludarabine monotherapy in chronic lymphocytic leukemia]. AB - Fludarabine has widely been studied in chronic lymphocytic leukemia (CLL), with impressive remission rates in refractory, relapsed or untreated disease. In our study the clinical response and survival of 9 patients with a mean follow-up time of 28 months after treatment with fludarabine as a single agent for CLL has been evaluated. Seven patients were previously treated. Partial remission was achieved in 8 patients. No complete remission was seen. The cytoreductive activity of fludarabine was excellent in all the 9 patients. The median time to progression was 13.5 months. The median survival time from entering the trial was 27.9 months. Our patients tolerated the fludarabine treatment extremely well. Although fludarabine has been established as the most active single agent in CLL, most patients will have recurrent disease. 7/9 patients relapsed in our study, and they were given further chemotherapy. Grade 4 hematologic toxicity was observed in 2 patients. During the fludarabine treatment the frequency of infections decreased, but in the following 12 months increased again. Three fludarabine treated patients developed high-grade non-Hodgkin lymphoma. It is concluded that fludarabine is a highly useful agent in CLL. PMID- 10463033 TI - [Autoradiography of neurotransmitter receptors in whole human brain hemisphere sections]. AB - Autoradiography is one of our most important tools to gain knowledge about neurotransmitter-receptors playing a key-role in information transmission between neurons. Autoradiography, in its most sophisticated form, is performed on whole human hemispheric sections. The main objective of the authors is to present this application of autoradiography. This in vitro method produces images with high spatial resolution that enable us to qualitatively and quantitatively characterize the regional distribution of the receptors under study. With this technique both the different receptor systems in various physiological and pathological conditions of the brain and the pharmacological parameters of the radioligand, itself, used for a given investigation can be analysed. As a consequence, the results of autoradiography can be successfully used in drug development and trial, brain research and, indirectly, in the every day practice of physicians (diagnosis, differentialdiagnosis, therapy). Autoradiography plays an important role in the validation of in vivo techniques (positron emission tomography, single photon emission tomography) and results in a more complex (in vivo and in vitro) insight into the neurochemical organisation of the brain. PMID- 10463035 TI - Annual meetings of the Canadian Association of General Surgeons and the Canadian Society of Colon and Rectal Surgeons. 68th Annual meeting of the Royal College of Physicians and Surgeons of Canada. Montreal, Quebec, Canada. September 23-26, 1999. Abstracts. PMID- 10463034 TI - [Neuromuscular choristoma]. AB - Neuromuscular choristomas are malformative pseudotumoral masses composed of striated muscle and peripheral nerves. This rare condition almost exclusively affects large nerve trunks of infants and young children, and may cause neurologic deficits of variable severity. We report a case of neuromuscular choristoma identified in the lumbosacral lipoma of a 4-month-old boy. The lesion was characterized by an organoid association of myogenic and neurogenic elements reminiscent of neuromuscular units. Myosin immunophenotyping revealed disordered groups of type-I and type-II extrafusal myofibers. There was no immunoreactivity for smooth muscle specific alpha actin. Most participating axons were of the large myelinated type. Pathogenetic theories of neuromuscular choristoma involve aberrant migration of developing peripheral nerves, dysgenesis of muscle spindles, as well as mesenchymal differentiation of the primitive neuroectoderm. Neuromuscular choristomas arising in connection with congenital lipomas may derive from a dysembryogenic process of stem cells of the caudal neural tube. PMID- 10463036 TI - 23rd International Epilepsy Congress. Prague, Czech Republic, September 12-17, 1999. Abstracts. PMID- 10463037 TI - European Federation of National Associations of Orthopaedics and Traumatology. Brussels Congress. June 1999. Abstracts. PMID- 10463038 TI - The expert consensus guideline series. Treatment of Posttraumatic Stress Disorder. The Expert Consensus Panels for PTSD. PMID- 10463039 TI - 18th Joint meeting of the British Endocrine Societies. Bournemouth, 12-15 April 1999. Abstracts. PMID- 10463040 TI - Annual meeting of the American Academy of Otolaryngology--Head and Neck Surgery Foundation, Inc. New Orleans, Louisiana, USA. September 26-29, 1999. Abstracts. PMID- 10463041 TI - Antibiotic prescribing rates. PMID- 10463042 TI - Antibiotic prescribing rates. PMID- 10463043 TI - "I need more power, Scotty". PMID- 10463044 TI - Detecting adverse drug reactions. PMID- 10463045 TI - Higher neonatal morbidity after routine early hospital discharge: are we sending newborns home too early? AB - BACKGROUND: A growing body of evidence suggests that the trend toward earlier discharge may affect newborn morbidity. The authors assessed how hospital readmission rates were affected by a clinical guideline aimed at discharging newborns from hospital 24 hours after birth. METHOD: A retrospective before-after cohort study was conducted involving 7009 infants born by uncomplicated vaginal delivery at a large level II hospital in Toronto between Dec. 31, 1993, and Sept. 29, 1997. The primary outcome was a comparison of the rate of hospital readmission among newborns before (5936 infants) and after (1073 infants) the early-discharge policy was implemented (Apr. 1, 1997). The causes for readmission were secondary outcomes. RESULTS: Before the early-discharge guideline was implemented, the mean length of stay declined from 2.25 days (95% confidence interval [CI] 2.18-2.32) to 1.88 days (95% CI 1.84-1.92) (p < 0.001). After implementation there was a further decline, to 1.62 days (95% CI 1.56-1.67) (p < 0.001). A total of 126 infants (11.7%) in the early-discharge cohort required readmission by 1 month, as compared with 396 infants (6.7%) in the preguideline cohort (odds ratio 1.86, 95% CI 1.51-2.30). The main reason for early readmission was neonatal jaundice, with a higher rate among infants in the early-discharge cohort than among those in the preguideline cohort (8.6% v. 3.1%; odds ratio 2.96, 95% CI 2.29-3.84). INTERPRETATION: Decreases in newborn length of stay may result in substantial increases in morbidity. Careful consideration is needed to establish whether a reduction in length of stay to less than 24 to 36 hours is harmful to babies. PMID- 10463046 TI - Reference-based pricing of prescription drugs: exploring the equivalence of angiotensin-converting-enzyme inhibitors. AB - BACKGROUND: Reference-based pricing is a cost-containment policy applied to prescription drugs that are in the same class and deemed to be therapeutically equivalent. Recent reference-based pricing measures have targeted several drug classes, including angiotensin-converting-enzyme (ACE) inhibitors. The objective of this study was to assess whether patients treated for hypertension with various ACE inhibitors differed in their utilization of health care services and hence, whether the various ACE inhibitors should be considered therapeutically equivalent. METHODS: A retrospective cohort was formed from 4709 Saskatchewan residents aged 40-79 years who initiated treatment for hypertension with 1 of the 3 most frequently prescribed ACE inhibitors (captopril, enalapril or lisinopril) between Jan. 1, 1991, and Dec. 31, 1993. Information obtained from universal insurance databases included prescription drug use, the number of visits to a general practitioner (GP) or specialist and the number of hospital admissions during the year before treatment was initiated and during a follow-up period of up to 4 years. Rates were statistically adjusted for potential confounding variables and compared across treatment groups. RESULTS: Of the 4709 patients, 529 were prescribed captopril initially, 2939 enalapril and 1241 lisinopril. After treatment was initiated patients prescribed captopril were dispensed more medications on average, with an overall rate of 18.6 prescriptions per patient per year (v. 16.4 and 14.7 for enalapril and lisinopril users respectively); they were admitted to hospital more often, and they made more visits to GPs and specialists. The adjusted rate ratio of the number of visits to a GP for patients receiving enalapril, relative to captopril, was 0.84 (95% confidence interval [CI] 0.80-0.88), and for those receiving lisinopril it was 0.79 (95% CI 0.74 0.83). The adjusted rate ratios for the number of visits to a specialist were similar but lower, and for the number of hospital admissions they were 0.82 for patients prescribed enalapril initially (95% CI 0.73-0.93) and 0.65 (95% CI 0.56 0.75) for those prescribed lisinopril. INTERPRETATION: Patients with hypertension who are initially prescribed captopril used health care services more than those initially prescribed enalapril or lisinopril. This suggests that ACE inhibitors may not be therapeutically equivalent. PMID- 10463048 TI - Evaluating the impact of reference-based pricing. PMID- 10463047 TI - Prevalence and predictors of white-coat response in patients with treated hypertension. AB - BACKGROUND: White-coat response, defined as higher office blood pressure readings than ambulatory readings, is common. Few studies have estimated its prevalence among subjects with treated hypertension, and almost none have defined its determinants. The objective of this study was to estimate the prevalence of white coat response among subjects with treated hypertension and to determine whether the phenomenon could be predicted using clinical and psychometric data. METHODS: A total of 103 treated patients (55 men and 48 women) with uncontrolled hypertension who attended a hypertension outpatient clinic in Saskatoon between September 1993 and December 1995 were entered into the study. Patients had at least 2 clinic blood pressure readings of 140/90 mm Hg or higher, had no target organ damage or left ventricular hypertrophy, and had been prescribed 2 or more classes of antihypertensive drugs. Patients had blood pressure measured in the supine position in the clinic, were placed on 24-hour ambulatory blood pressure monitoring and then completed questionnaires before returning to the clinic. Patients were classified as exhibiting a white-coat response if their mean daytime ambulatory systolic/diastolic blood pressure was 139/89 mm Hg (both) or less, or if the systolic/diastolic pressure was at least 20/15 mm Hg (both) lower than the clinic reading. RESULTS: Eleven men (20%, 95% confidence interval [CI] 10%-33%) and 26 women (54%, 95% CI 39%-69%) showed white-coat response. Logistic regression modelling showed that determinants such as stress had significantly different effects among men and women. Separate models were therefore created for men and women. For women, perceived level of stress was the most important predictor of white-coat response (odds ratio [OR] per unit 7.0, 95% CI 1.3-36.0), followed by time since diagnosis. For men, depression was a weak predictor, with higher depression scores predicting sustained hypertension (OR per unit 1.2, 95% CI 1.01-1.5). INTERPRETATION: Sex is an important factor in white-coat response. Attempts to predict white-coat response from psychometric variables should take sex differences into account. Clinical variables were not effective as predictors of white-coat response. PMID- 10463050 TI - Accurate blood pressure measurement: why does it matter? PMID- 10463049 TI - Withholding treatment in white-coat hypertension: wishful thinking. PMID- 10463051 TI - Oncology Rehabilitation Program at the Ottawa Regional Cancer Centre: program description. AB - The Ottawa Regional Cancer Centre offers an Oncology Rehabilitation Program to patients with cancer. Between January 1997 and December 1998, 254 patients with cancer participated in the program. This paper describes the program and its participants. The program's strengths, limitations and future directions are also discussed. PMID- 10463052 TI - Evaluating reference-based pricing: initial findings and prospects. AB - Reference-based pricing is a controversial policy mechanism used to control pharmaceutical expenditures. After its implementation in some European countries, the British Columbia government introduced a version of reference-based pricing in October 1995. The authors reviewed previous studies of reference-based pricing in other countries and conducted a preliminary assessment of the impacts of the BC system by analysing secondary utilization and cost data. After the introduction of reference-based pricing in other jurisdictions within the Organisation for Economic Cooperation and Development, there was a temporary reduction in the rate of growth of total pharmaceutical expenditures, followed by a return to previous growth trends in subsequent years. Similarly, initial data from BC showed dramatic declines in annual expenditures for drugs within referenced categories (from $42.0 million the year before reference-based pricing was introduced to $23.7 million the year after). Although early evidence suggests that reference-based pricing in BC is indeed reducing drug expenditures, much more research is needed to make a final determination of its success. A more comprehensive and longitudinal evaluation of reference-based pricing is needed and should take into account a wide range of non-cost impacts, the most important of which are the effects on health outcomes. PMID- 10463053 TI - Edmonton Regional Palliative Care Program: impact on patterns of terminal cancer care. AB - The Edmonton Regional Palliative Care Program was established in July 1995 to measure the access of patients with terminal cancer to palliative care services, decrease the number of cancer-related deaths in acute care facilities and increase the participation of family physicians in the care of terminally ill patients. In this retrospective study the authors compared the pattern of care and site of deaths before establishment of the program (1992/93) and during its second year of operation (1996/97). Significantly more cancer-related deaths occurred in acute care facilities in 1992/93 than in 1996/97 (86% [1119/1304] v. 49% [633/1279]) (p < or = 0.001). The number of inpatient days decreased, from 24,566 in 1992/93 to 6960 in 1996/97. More cancer patients saw a palliative care consult team in 1996/97 than in 1992/93 (82% v. 22%). The shift from deaths in acute care facilities to palliative hospices suggests that the establishment of an integrated palliative care program has increased access of patients with terminal cancer to palliative care. PMID- 10463054 TI - Move to market gene pool angers Iceland's MDs. PMID- 10463055 TI - Billing dispute infuriates Montreal ER doctors, sparks mass-resignation threat. PMID- 10463056 TI - Identification of a Drosophila prolyl endopeptidase and analysis of its expression. AB - Prolyl endopeptidases (PEPs) are believed to be involved in the metabolism of neuropeptide hormones (reviewed in Mentlein [1988]). Genes encoding PEPs have been isolated from various species, but their expression patterns during development have not been determined. In this study, we isolated a gene encoding a predicted PEP from the fruitfly Drosophila melanogaster. The gene encodes a predicted 756-amino acid protein having extensive sequence similarity to human PEP. We demonstrated that the Drosophila gene (DPEP) is expressed in a spatially restricted pattern in imaginal discs and the larval brain. Our results suggest a role for DPEP in the regional specification of larval tissues. They also provide a starting point for a genetic analysis of the function of this enzyme during development. PMID- 10463058 TI - Increase in the immunogenicity of HIV peptide antigens by chemical linkage to polytuftsin (TKPR40). AB - The use of synthetic peptide antigens in human prophylaxis still suffers from the very important problem of finding suitable carriers devoid of side effects. A desirable carrier for use in humans would be poorly immunogenic by itself, yet it would enhance the immune response to the peptide antigen. In the study reported herein, we examined the role of polytuftsin (TKPR40), a synthetic polymer of the natural immunomodulator tuftsin, as a carrier for synthetic peptides of HIV derived from the gp41 and gp120 proteins. Chimeric immunogens were constructed by chemical linkage between synthetic peptides of HIV and polytuftsin. These were employed for immunization of mice of different MHC haplotypes, and the humoral and cellular immune responses developed against the peptides were assessed by measuring total IgG, IgG, subclasses, T-cell proliferation, and in vitro cytokine release. A significantly stronger immune response was observed in mice immunized with the peptide-polytuftsin conjugates than in mice receiving the peptide dimers (peptide-peptide). Peptide-polytuftsin conjugates induced IgG2a and IgG2b isotype switching after both primary and secondary immunization. In addition, there was a positive correlation between the amounts of cytokines and the shift in the IgG isotypes. These data suggest that the use of polytuftsin as a carrier may increase the immune response against poorly immunogenic synthetic peptides. PMID- 10463057 TI - Targeting of hepatitis C virus core protein for MHC I or MHC II presentation does not enhance induction of immune responses to DNA vaccination. AB - We analyzed different vaccine approaches aimed at enhancing CD4(+)- and CD8(+) dependent responses against hepatitis C virus (HCV) core antigen. Specific DNA vectors expressing various forms of the core in fusion with the ubiquitin or the lysosome-associated membrane protein (LAMP) were generated. These expressed the full-length wildtype core; the full-length core expressed as a covalent fusion with the ubiquitin; the full-length core expressed as a noncovalent fusion with the ubiquitin and containing a N-stabilizing or N-destabilizing residue; and the full-length core expressed as a fusion with the LAMP sequence. In vitro expression levels of the different plasmids differed by as much as tenfold. After injection into mice, none of the plasmids yielded a detectable antibody response, whereas core-specific cytotoxic T-lymphocyte (CTL) activity could be observed with all plasmids as long as 21 weeks postimmunization. No increase in CTL activity (ranging from 7% to 34% specific lysis) was observed with the ubiquitin fusion-expressed core antigens compared with the wildtype core. The lowest CTL activity (< 5% specific lysis) was observed with the LAMP fusion. This vector was nonetheless unable to induce a detectable proliferative response. Screening of 10 different putative CTL peptide epitopes failed to reveal newly targeted epitopes when the core-fusion plasmids were used compared with the wildtype core expressing plasmid. These data underline the difficulty in optimizing anti-core cellular immune response using molecular targeting strategies in DNA-based vaccination. PMID- 10463059 TI - Differential activity of the -2.7 kb chicken lysozyme enhancer in macrophages of different ontogenic origins is regulated by C/EBP and PU.1 transcription factors. AB - Expression of the chicken lysozyme gene is upregulated during macrophage maturation. Recently, an additional regulatory feature was discovered: the gene is differentially expressed in macrophages of embryonic/fetal and adult origin. The lysozyme gene is only weakly expressed in mature embryo-derived macrophages, whereas there is a high level of expression in macrophages derived from adult animals. This finding provided a molecular tool to investigate the heretofore ill defined differences between embryonic/fetal- and adult-type macrophages. We showed that the low expression in the embryo is associated with reduced activity of the myeloid-specific -2.7 kb lysozyme enhancer. Our protein-binding analyses and transfection studies demonstrated that this enhancer, in order to be fully active in activated macrophages, requires the combined action of C/EBPs, PU.1, and a third, as yet unidentified, protein binding to an AP-1-like site. Of these three, PU.1 and C/EBPs display significantly reduced nuclear DNA-binding activities in embryo-derived macrophages compared with adult-type cells. These results point to different roles of C/EBPs and PU.1 in embryonic/fetal and adult myelopoiesis. PMID- 10463060 TI - Decreased expression of manganese superoxide dismutase in transformed cells is associated with increased cytosine methylation of the SOD2 gene. AB - Tumor cells express lower levels of manganese superoxide dismutase (MnSOD) than their normal counterparts. Enforced expression of MnSOD reverses the malignant phenotype of many transformed cells, suggesting that SOD2 is a tumor suppressor. The SOD2 gene contains a large CpG island spanning > 3.5 kb that starts near the 5' edge of the promoter and extends into intron 2. We hypothesized that the difference in SOD2 expression between tumor cells and their normal cell counterparts might be secondary to differences in their cytosine methylation patterns in this CpG island. To test this hypothesis, we analyzed the methylation status of the SOD2 gene in two cell line models that show differential MnSOD expression between normal and SV40-transformed cells: WI38 and MRC5 and their SV40-transformed variants, WI38-VA and MRC5-VA. We subdivided the SOD2 gene CpG island into 10 individual regions for analysis by bisulfite genomic sequencing. A region located in intron 2 displayed a significant increase in cytosine methylation in both transformed cell lines that expressed low levels of MnSOD mRNA compared with their normal cell counterparts. Recent studies by others have shown that SOD2 intron 2 is a potent transcriptional enhancer. The association between increased cytosine methylation of the SOD2 intron 2 region and decreased MnSOD expression in transformed cells compared with their normal counterparts suggests that an epigenetic mechanism contributes to the differential SOD2 gene expression between these normal and SV40-transformed cells. PMID- 10463061 TI - The product of the primary response gene BRF1 inhibits the interaction between 14 3-3 proteins and cRaf-1 in the yeast trihybrid system. AB - The 14-3-3 proteins are small abundant cytosolic eukaryotic proteins that associate with and modulate the activity of numerous other proteins. The 14-3-3 beta isoform has been shown to bind to the product of the protooncogene cRaf-1 and to facilitate its activation by Ras. Using the yeast two-hybrid system, we have demonstrated that 14-3-3 beta and another isoform, 14-3-3 tau, bind to the product of the primary response gene BRF1 and that the interaction between each isoform and BRF1 is significantly stronger than that with cRaf-1. We further demonstrated that the charge of residue 187 in 14-3-3 beta regulates its affinity for both BRF1 and cRaf-1. The interaction of either isoform with BRF1 requires both proteins to be fully intact. When all three proteins are coexpressed in a yeast trihybrid system, BRF1 interferes significantly with the binding of 14-3-3 to full-length cRaf-1 as well as to its regulatory and kinase domains. Using quantitative reverse transcription-polymerase chain reaction, 14-3-3 beta and BRF1 were found to be coexpressed in four different human tissues, suggesting a biologic role for their interaction in the regulation of cRaf-1-mediated signal transduction processes. PMID- 10463062 TI - The place of culture in DSM-IV. AB - This paper critically reviews the process and outcome of an effort to enhance the cultural validity of DSM-IV and outlines recommendations to improve future diagnostic systems. An ordered presentation of the antecedents and the main phases of this developmental effort is followed by a content analysis of what was proposed and what was actually incorporated, and a conceptual analysis of underlying biases and their implications. The cultural effort for DSM-IV, spearheaded by a scholarly independent NIMH workgroup, resulted in significant innovations including an introductory cultural statement, cultural considerations for the use of diagnostic categories and criteria, a glossary of culture-bound syndromes and idioms of distress, and an outline for a cultural formulation. However, proposals that challenged universalistic nosological assumptions and argued for the contextualization of illness, diagnosis, and care were minimally incorporated and marginally placed. Although a step forward has been taken to introduce cultural elements in DSM-IV much remains to be done. Further culturally informed research is needed to ensure that future diagnostic systems incorporate a genuinely comprehensive framework, responsive to the complexity of health problems in increasingly multicultural societies. PMID- 10463063 TI - Clinical relevance of contemporary cultural psychiatry. AB - In recent years, the field of cultural psychiatry has gained recognition and accumulated evidence of its clinical relevance. This article examines the intersections of culture and psychopathology and describes five independent but interrelated clinical dimensions that identify and define culture as: a) an interpretive/explanatory tool, b) a pathogenic/pathoplastic agent, c) a diagnostic/nosological factor, d) a therapeutic/protective element, and e) a service/management instrument. Along these lines, conceptual boundaries, clinical findings, specific applications, and research implications for each of the five dimensions are systematically reviewed. Cultural psychiatry adds significantly to the comprehensiveness of psychiatric evaluation and management and addresses prominent issues regarding understanding, classification, diagnosis, and competent treatment of most psychiatric disorders in every society and region of the world. Based on the strength of these clinical dimensions, and on the related educational and research efforts, cultural psychiatry can also contribute decisively to the design of comprehensive mental health policies. PMID- 10463064 TI - Psychopathological characteristics of recovered bulimics who have a history of physical or sexual abuse. AB - We sought to clarify the influence of a history of sexual or physical abuse on a variety of psychopathologies in subjects with bulimia nervosa (BN). To avoid confounding effects, the presence of a history of sexual or physical abuse, lifetime axis I disorders, and personality disorders were assessed through direct structured interviews in 44 subjects recovered from BN for at least 1 year. Twenty abused subjects (45%) were significantly more likely than 24 subjects without abuse to have severe general psychopathology and eating disturbance. Compared with nonabused subjects, abused subjects showed a trend toward more frequent lifetime diagnoses of posttraumatic stress disorder and substance dependence. These results suggest that abusive experiences may be associated with some psychopathology of BN, particularly related to anxiety, substance abuse, and more severe core eating disorder pathology. PMID- 10463065 TI - Concurrent and predictive validity of antisocial personality disorder subtyping among substance abusers. AB - Three hundred seventy inpatient and outpatient substance abusers were divided according to presence and subtype of antisocial personality disorder (APD) into groups comparing: a) adult antisocial behavior (AAB) versus full APD; b) APD with low versus high sociopathy; c) APD with versus without lifetime depression; and d) APD with versus without other axis II disorders. Multivariate regression was used to predict the unique contribution to the variance in baseline and 12-month follow-up measures of substance use, psychiatric severity, and personality. The presence of comorbid axis II pathology was the strongest predictor of baseline severity in all three domains. APD substance abusers with lifetime depression exhibited greater baseline to follow-up reductions in psychiatric severity than those APD substance abusers without a history of depression. All APD subtypes improved over time with treatment, suggesting that this diagnosis does not necessarily indicate poor prognosis. PMID- 10463066 TI - Attention deficit hyperactivity disorder and treatment outcome in opioid abusers entering treatment. AB - Symptoms of DSM-IV attention-deficit hyperactivity disorder (ADHD) were determined in patients entering methadone maintenance treatment. The relationship of ADHD to psychiatric and substance abuse comorbidity, attention testing, and treatment outcome was analyzed; 19% of patients had a history of ADHD, and 88% of these had current symptoms. Continuous Performance Testing indicated poorer attention in patients with ADHD. The only substance use disorder more common in the ADHD group was clonidine. There was significantly more current axis I, dysthymic disorder, anxiety disorder (including social phobia), and antisocial personality disorder in the ADHD patients. There was no difference between groups at the 1-year follow-up for illicit drug use, treatment retention, or treatment performance. The ADHD diagnosis did not convey significant prognostic implications for methadone maintenance treatment. A strong psychiatric assessment and treatment focus in the treatment program may help to explain the good treatment outcome. PMID- 10463067 TI - The association of medical comorbidity in schizophrenia with poor physical and mental health. AB - This study determined the prevalence of medical comorbidities in a cohort of persons receiving treatment for schizophrenia and the association of medical comorbidity with physical and mental health status. A total of 719 persons with schizophrenia sampled from a variety of community and treatment settings as part of the schizophrenia Patient Outcomes Research Team (PORT) participated in a survey interview. Multiple regression analyses were used to assess sociodemographic factors associated with the number of current medical comorbidities and the association of medical comorbidity count with patient ratings of physical health, mental health, symptoms, and quality of life. The majority of patients reported at least one medical problem. Problems with eyesight, teeth, and high blood pressure were most common. A greater number of current medical problems independently contributed to worse perceived physical health status, more severe psychosis and depression, and greater likelihood of a history of a suicide attempt. This study underscores the need to attend to somatic health care for persons with schizophrenia as well as the linkage of physical and mental health status. PMID- 10463068 TI - Coping with Parkinson's disease and refractory epilepsy. A comparative study. AB - Coping strategies and their correlations with demographic and illness related data, depression, locus of control, and psychosocial adaptation were investigated in 45 patients with Parkinson's disease and 40 patients with intractable epilepsy. Three standardized self-report questionnaires were applied: the Freiburg Questionnaire of Coping with Illness (FKV), the von Zerssen Depression Scale (D-S), and the IPC-questionnaire measuring generalized locus of control beliefs. The Social Interview Schedule (SIS), a semi-structured interview, was used to measure psychosocial adaptation. Active, problem-focused and compliance strategies were predominantly used and regarded as most helpful in both groups of patients. Hence, Parkinson and epilepsy patients used similar coping patterns despite the strong dissimilarities of symptoms and illness-associated burdens. The level of depression was not significantly different in both groups and in the range of other chronic somatic diseases. The use of coping patterns, which are regarded as maladaptive, was correlated with distinct depression and a poor psychosocial adaption. In conclusion, no indications for illness-specific coping patterns were found in patients with Parkinson's disease and epilepsy. In both groups, certain coping strategies are associated with good and others with poor psychosocial adjustment. These results indicate the possibility of improving psychosocial adjustment by supporting effective strategies. PMID- 10463069 TI - Influence of personality disorders on therapy outcome in somatoform disorders at 2-year follow-up. PMID- 10463070 TI - Response to depression and anxiety Stroop stimuli in posttraumatic stress disorder, obsessive-compulsive disorder, and major depressive disorder. PMID- 10463071 TI - Biophysical studies on ATP synthase. AB - The isolation of ATP synthase (F0F1) (82) and F0 (83) 34 years ago finally revealed that F0F1 is a motor composed of F0 (ion-motor, abc subunits) and F1 (ATP-motor, alpha 3 beta 3 gamma delta epsilon subunits) (Fig. 1). The single molecule videotape (4, 5, 65, 66) revealed that gamma epsilon axis of F1 rotates counterclockwise, proceeds by each 2 pi/3 step, and is driven by torque of 42 pN.nm (12) with nearly 100% efficiency (5) (Fig. 4). The motor is composed of a rotor (gamma epsilon-F0-c) and a stator (alpha 3 beta 3 delta-F0-ab), and the rotor is connected to a shaft (gamma epsilon). Since F0F1 is driven by delta microH+ (9, 10, 84), biophysical studies on stable TF0F1 (1, 7) are essential to elucidate the mechanism. These include nanomechanics (4, 5) (Fig. 4), crystallography (2, 3) (Figs. 2 and 3), NMR (51, 52), ESR (56), synchrotron analysis (3, 28), and electrophysiology (10, 25). The KmATP value of rotation is 0.8 microM, with the Vmax of 3.9 rps (5). This corresponds to the bi-site catalysis in proton transport by F0F1 (10, 70, 84). X-ray crystallography of MF1 (2) and the alpha 3 beta 3 oligomer of TF1 (3) (Fig. 2) together with mutation analyses revealed the role of residues in the rotation. The idea of elastic energy store is proposed in alpha 3 beta 3 gamma during the stepping time (up to a few sec) after the ATP binding. Biological studies have partially clarified the genetic and kinetic regulation of the rotation in MF1. Both theories (6, 7, 62, 64, 85) and the biological significance (17) of the intramolecular rotation of F0F1 await further studies, especially those of F0 and minor subunits. PMID- 10463072 TI - Ryanodine receptor isoforms in excitation-contraction coupling. AB - Three genomically distinct isoforms of RyR are now known. RyR1 homologue is the primary isoform in skeletal muscles, whereas in cardiac muscles it is RyR2 homologue. RyR3 homologue occurs ubiquitously in many cells, but the biological function is little known, partly because of its minuscule amount in mammalian cells. The difference among RyR isoforms may not be so great in CICR activity, in other words, in the interaction of RyR isoforms with Ca2+, adenine nucleotides and caffeine. Species specificity among RyR1 homologues may be more important in the apparent difference between RyR1 and RyR3 homologues. CICR is likely to be the dominant underlying mechanism for E-C coupling in the cardiac muscle and probably in cells other than the skeletal muscle where the significance of CICR is controversial in physiological contraction. In E-C coupling of skeletal muscle (DICR), the reciprocal tight interactions between DHPR and RyR1 are critically required. The alpha 1 subunit of DHPR was only the main target of our current interests in the interaction with RyR1; the involvement of auxiliary subunits of alpha 2/delta and beta subunits and their mutual interactions, however, are also important. DICR and CICR in RyR1 share common properties of stimulation by concentrated solutes and modulation by luminal calcium or Ca2+, suggesting that the main difference between the two Ca2+ release mechanisms may be in the gating mechanism of the channel. Further investigations are required to understand molecular interactions during E-C coupling. PMID- 10463073 TI - Evolution of the carabid ground beetles. AB - The phylogenetic relationships of the carabid ground beetles have been estimated by analysing a large part of the ND5 gene sequences of more than 1,000 specimens consisting of the representative species and geographic races covering most of the genera and subgenera known in the world. From the phylogenetic analyses in conjunction with the mtDNA-based dating, a scenario of the establishment of the present habitats of the respective Japanese carabids has been constructed. The carabid diversification took place ca. 40 MYA as an explosive radiation of the major genera. During evolution, occasional small or single bangs also took place, sometimes accompanied by parallel morphological evolution in phylogenetically remote as well as close lineages. The existence of silent periods, in which few morphological changes took place, has been recognized during evolution. Thus, the carabid evolution is discontinuous, alternatively having a phase of rapid morphological change and a silent phase. PMID- 10463074 TI - Induced potential model of muscular contraction mechanism and myosin molecular structure. AB - The proposed model is characterized by the constant r (Eq. 2-1), the induced potential (Fig. 1), two attached states of a myosin head (Fig. 1), the nonlinear elastic property of the crossbridge (Eq. 2-7), and the expression of U* (Eqs. 3-8 and 3-9), which led us to the following conclusions. 1. The following various magnitudes of myosin head motion are compatible with each other: about 2 nm of the quantity called power stroke by Irving (27), which is the mean moving distance of myosin head in the isometric tension in our model, 4-5 nm of the displacement of a single myosin head during one ATP hydrolysis cycle (Molloy et al. (20)) or a few tens of nm when the actin and myosin filaments are set parallel (Tanaka et al. (21) and Kitamura et al. (42)), and more than 200 nm of the myosin head displacement in a multi-myosin head system below 22 degrees C (Harada et al. (19)). 2. There is one-to-one coupling between the ATP hydrolysis cycle and the attachment-detachment cycle of a myosin head in accordance with the generally accepted concept of chemical reactions, since the head is trapped in the spatially shifting wide potential well (Fig. 1) until epsilon ATP is exhausted. Here, an actin filament interacts with a myosin head like a single molecule. 3. The calculated tension dependence of muscle stiffness agrees well with the observations by Ford et al. (12), as shown in Fig. 9. 4. The calculated shortening velocity V of muscle as a function of P/P0 agreed very well with experimental results as shown in Fig. 13. The deviation from the Hill equation (34) observed by Edman (32) is related with U* being effectively infinite for f1 < kappa b yc0 (Fig. 10). 5. Calculated energy liberation rate W + H as a function of P/P0 has characteristics almost the same as the Hill equation (33), and agrees well with the experimental results as shown in Fig. 14. 6. The time course of tension recovery after a quick length change is determined by four parameters: kappa f, kappa b, a, and Z0. Among them, kappa f, kappa b (Eq. 2-22) and a (Eq. 4 21) are readily determined by analysis of the steady filament sliding and p0. Calculations of T1/T0 and T2/T0 with these three parameters are in very good agreement with experimental data (Fig. 21). Calculated tension variations by assigning the value in Eq. 4-23 to Z0 agree with the observation (Fig. 17). 7. The model suggests that large fluctuations exist in relative positions between the actin and myosin filaments even when the load on a muscle is kept constant (Fig. 23). Taking this fluctuation into account, the time course of the isotonic velocity transient shown in Fig. 22 becomes understandable referring to Fig. 24. 8. The experimental data of the delta yhs vs. delta P/P0 relationship (Fig. 25) is explained. The delta yhs value at delta P/P0 = 0 (about 5 nm) supports the two attached-state model and thus indicates that the incremental unit step of a myosin head motion along an actin filament is close to L (5.46 nm). PMID- 10463075 TI - Multiple sequence alignment: algorithms and applications. AB - Elucidation of interrelationships among sequence, structure, function, and evolution (FESS relationships) of a family of genes or gene products is a central theme of modern molecular biology. Multiple sequence alignment has been proven to be a powerful tool for many fields of studies such as phylogenetic reconstruction, illumination of functionally important regions, and prediction of higher order structures of proteins and RNAs. However, it is far too trivial to automatically construct a multiple alignment from a set of related sequences. A variety of methods for solving this computationally difficult problem are reviewed. Several important applications of multiple alignment for elucidation of the FESS relationships are also discussed. For a long period, progressive methods have been the only practical means to solve a multiple alignment problem of appreciable size. This situation is now changing with the development of new techniques including several classes of iterative methods. Today's progress in multiple sequence alignment methods has been made by the multidisciplinary endeavors of mathematicians, computer scientists, and biologists in various fields including biophysicists in particular. The ideas are also originated from various backgrounds, pure algorithmics, statistics, thermodynamics, and others. The outcomes are now enjoyed by researchers in many fields of biological sciences. In the near future, generalized multiple alignment may play a central role in studies of FESS relationships. The organized mixture of knowledge from multiple fields will ferment to develop fruitful results which would be hard to obtain within each area. I hope this review provides a useful information resource for future development of theory and practice in this rapidly expanding area of bioinformatics. PMID- 10463076 TI - Detection of proteins binding to short RNA.DNA hybrids or short antisense oligonucleotides in Xenopus laevis oocytes and human macrophage cell extracts by photoaffinity radiolabeling. AB - Using a 12 base pair RNA.DNA hybrid, substituted with bromouracil on either the RNA or DNA strand, we have detected by photoaffinity radiolabeling a limited set of proteins able to bind to RNA.DNA hybrids in both Xenopus oocyte extracts and human macrophage extracts. Resulting patterns of crosslinked proteins were highly dependent on the strand (DNA or RNA) that was substituted. With one exception, none of the proteins investigated in competition experiments was found to be absolutely specific for RNA.DNA hybrids, as at least one other nucleic acid, either single-stranded DNA or single-stranded RNA, was found to compete efficiently. None of the proteins detected in this assay correspond to the size expected for RNases H. Using the same methodology, we have detected proteins that bind to short oligodeoxyribonucleotides. Although we have essentially detected in Xenopus oocytes one prominent protein of approximately 75 kDa, corresponding to replication protein A (RPA) whatever the oligonucleotide used, the patterns obtained with extracts of human macrophages were more complex and dependent on the oligonucleotide used. If a protein corresponding to RPA was observed most of the time, other crosslinks of similar or sometimes higher intensity were also detected. Interestingly, among these, one protein of 35 kDa appears paradoxically to bind and crosslink to a dodecamer but not to an octadecamer containing the same sequence placed either at its 3'-end or 5'-end. PMID- 10463077 TI - Antisense technologies have a future fighting neurodegenerative diseases. AB - Our growing understanding of the role that unfavorable patterns of gene expression play in the etiology of neurodegenerative disease emphasizes the need for strategies to selectively block the biosynthesis of harmful proteins in the brain. Antisense technologies are ideally suited to this purpose. Tailor-designed to target specific RNA, antisense oligonucleotides and ribozymes offer tools to suppress the production of proteins mediating neurodegeneration. Although technical limitations must still be overcome, the antisense approach represents a novel and exciting strategy for intervention in diseases of the central nervous system. PMID- 10463078 TI - Therapeutic efficacy of an adenovirus-mediated anti-H-ras ribozyme in experimental bladder cancer. AB - Ras oncogenes are thought to play a critical role in cellular proliferation and tumorigenesis. Reversal of the malignant phenotype, inhibition of tumor growth, and decreased tumorgenicity have been demonstrated with the use of anti-H-ras ribozymes. In this study, the therapeutic efficacy of a hammerhead ribozyme targeting the mutated H-ras oncogene was investigated in an experimental bladder cancer model using a recombinant adenovirus as delivery vehicle. Tumors were established in nude mice by subcutaneous injection of EJ human bladder carcinoma cells harboring a point mutation of the H-ras gene. The tumors were treated with intralesional injections of an adenovirus expressing an anti-H-ras ribozyme (rAd Hras Rz) by different schedules at serial titers, and the tumor inhibition efficacy was analyzed. The viral infection efficacy and kinetics of ribozyme expression were also evaluated. Intralesional injection of rAd-Hras Rz resulted in significant antineoplastic effects in a dose-dependent fashion. Complete regression of the tumor was achieved by rAd-Hras Rz in several cases without recurrence during the 50-day observation period. Although there was moderate vector-associated cytotoxicity in this cell line, complete regressions were not observed in the cases treated with control adenovirus vectors or vectors expressing an inactive anti-H-ras ribozyme or anti-H-ras antisense oligonucleotides. These results suggest the efficacy of a ribozyme-encoding adenovirus in the experimental gene therapy of human bladder cancer. PMID- 10463079 TI - An antisense oligonucleotide primer. PMID- 10463081 TI - Effectiveness of donepezil in treating Alzheimer's disease. PMID- 10463080 TI - Fighting the battle against breast cancer. So close and yet so far. PMID- 10463082 TI - Are there benefits to treating these conditions? PMID- 10463083 TI - Congenital rubella syndrome. How can we do better? AB - QUESTIONMy 22-year-old patient immigrated to Canada 6 years ago from a country where there are no routine immunizations. She is now at 10 weeks' gestation and was exposed to a child with rubella infection. Her immune status is unknown. Should I give her passive or active immunization? How should I follow her pregnancy? And what should I tell her about the risk for her fetus?ANSWERIf the exposure took place less than 1 week ago, blood should be sent for rubella antibody titre. If results are positive for IgG, she is immune and there is no risk for the fetus. If a patient's immune status is unknown and the time of exposure is uncertain, serum samples for rubella antibodies should be obtained 3 weeks apart. Detection of rubella-specific IgM in a sample will confirm recent infection. Your patient should be counseled about the potential risk for her fetus and referred to high-risk pregnancy clinic for follow up. If seroconversion does not take place, she requires immunization immediately postpartum before discharge from hospital. PMID- 10463084 TI - Ophthaproblem. Ocular histoplasmosis syndrome. PMID- 10463085 TI - Dermacase. Keratosis piloris. PMID- 10463086 TI - Each of the following statements about human immunodeficiency virus (HIV) in pregnancy is true. PMID- 10463088 TI - Practice tips. A method for presenting maternal serum screening. Promoting choice and saving time. PMID- 10463089 TI - Does bed rest improve any of the important outcomes of sciatica? PMID- 10463090 TI - Who has screening mammography? Results from the 1994-1995 National Population Health Survey. AB - OBJECTIVE: To determine the characteristics of Canadian women aged 35 to 49 who receive screening mammograms not recommended by the Canadian Task Force on the Periodic Health Examination. DESIGN: Secondary data analysis of the 1994-1995 National Population Health Survey. SETTING: Patients' homes. PARTICIPANTS: From a full national representative sample of 17,626 Canadian residents, we selected 2053 women aged 35 to 49 with no breast problems. MAIN OUTCOME MEASURES: Age, education, employment status, marital status, immigrant status, region of residence, self-reported health status, having a regular doctor, smoking status, alcohol consumption, and having a confidant. RESULTS: Of the 2053 women in the sample, 825 (40.2%) had had a screening mammogram as part of a regular medical checkup; 1228 (59.8%) had never had one. Logistic analysis showed that respondents who were approaching age 50, had higher incomes, lived in Quebec, and had regular medical doctors were more likely to have screening mammograms. Statistical trends indicated that heavy drinkers were less likely and immigrants more likely to have mammograms (not significant at P < .01: P = .012 and P = .02, respectively). CONCLUSIONS: Most of these findings are consistent with those of other studies of women 50 and younger. The findings suggest that the patient variables associated with having mammograms in those younger than 50 might be similar to those in women older than 50. An important next step is to determine whether this pattern of use has more to do with younger patients' demand for screening or with physicians' ordering of tests. Further research is also needed to understand the dynamics of the doctor-patient relationship in this situation. PMID- 10463091 TI - Teaching family practice residents breast cyst aspiration. AB - OBJECTIVE: To conduct the first study of teaching family practice residents the technique of breast cyst aspiration (BCA) using the Toronto Breast Cyst Aspiration Model (TBCAM) in a workshop. To determine whether this training increases their likelihood of performing the procedure in the future. To discover how residents evaluate the workshop as a teaching method. DESIGN: Prospective parallel-group trial from August to November 1997. Baseline questionnaires were given to both experimental and control groups in August. The experimental group was given a 1.5-hour workshop and a postworkshop questionnaire. Both groups were then given 3-month follow-up questionnaires. SETTING: Family practice residency program at the University of Toronto. PARTICIPANTS: The experimental group consisted of 15 family practice residents from one university-affiliated hospital site. The control group consisted of 54 family practice residents selected from the remaining sites. MAIN OUTCOME MEASURES: Confidence score, likelihood of performing BCA, and effectiveness of the workshop rated on a Likert scale ranging from 1-low to 5-high; and knowledge of BCA rated from 0 to 7. RESULTS: Three months after the workshop, 62% of study subjects and 31% of controls were above the median confidence score of 3.2 (chi 2 3.4, P > .05); 76% of subjects but only 11% of controls were above the median knowledge score of 4 (chi 2 16.67, P < .001); and 75% of subjects and 34% of controls were above the median likelihood to perform score of 3 (chi 2 5.37, P < .05). Most (95%) workshop effectiveness scores were 4s or 5s. CONCLUSIONS: Workshop training resulted in higher confidence, greater knowledge, and more likelihood of performing BCA. The workshop using the TBCAM was evaluated as a highly effective way to teach BCA. PMID- 10463092 TI - Breast cancer treatment and chemoprevention. AB - OBJECTIVE: To outline modern principles of surgery, radiation therapy, and systemic treatment of breast cancer, and to review preliminary data on breast cancer prevention. QUALITY OF EVIDENCE: A MEDLINE search was conducted from 1966 to the beginning of 1999; most of the studies reviewed are randomized clinical trials. MAIN MESSAGE: Breast conservation surgery should be offered to all women with early breast cancer because studies demonstrate survival rates equivalent to those with mastectomy. If mastectomy is chosen, breast reconstruction should be offered. Most women with breast cancer are treated systemically with either chemotherapy or tamoxifen, or both, and mortality is substantially reduced. Data indicating that tamoxifen prevents breast cancer are promising; more studies with both tamoxifen and raloxifene are under way. All women should be strongly encouraged to enter clinical trials. CONCLUSIONS: Because many issues face women recently diagnosed with breast cancer, they often seek out family physicians as advisors to help negotiate a complex treatment path. The possibility of preventing breast cancer will undoubtedly raise questions among family members of women with breast cancer that should appropriately be answered and referred, if necessary, by family physicians. PMID- 10463093 TI - Management of palpable breast lumps. Consensus guideline for family physicians. AB - OBJECTIVE: To describe an approach to managing women who present with palpable breast lumps. QUALITY OF EVIDENCE: Databases were searched from 1990 to 1998 using the search terms breast lumps, breast diseases, and breast cysts. Bibliographies of the articles obtained were searched for further relevant titles. Most evidence on management of breast cysts was obtained from cohort studies. Evidence on family physicians' approach to managing breast lumps is based on a review of the 1998 Canadian consensus guidelines and a review of a 1998 consensus guideline by 12 University of Toronto surgical oncologists (U of T guidelines). MAIN MESSAGE: Family physicians can manage women presenting with breast lumps if they have skill in breast cyst aspiration. Most breast cysts can be cured in minutes, thus avoiding unwarranted anxiety and eliminating unnecessary additional investigations and referrals. Women presenting with solid lesions should be referred to a surgeon. CONCLUSIONS: Breast cyst aspiration is a simple technique family physicians can use to either cure breast lumps or define appropriate cases for referral. PMID- 10463094 TI - Breast cancer in Canada. PMID- 10463095 TI - Obsessive compulsive disorder. PMID- 10463096 TI - Ethical approach to genital examination of children. Community Paediatrics Committee, Canadian Paediatric Society. PMID- 10463097 TI - Dynamic regulation of [Ca2+]i by plasma membrane Ca(2+)-ATPase and Na+/Ca2+ exchange during capacitative Ca2+ entry in bovine vascular endothelial cells. AB - The dynamic regulation of Ca2+ extrusion by the plasma membrane Ca(2+)-ATPase (PMCA) and Na+/Ca2+ exchange (NCX) was investigated in single cultured calf pulmonary artery endothelial (CPAE) cells using indo-1 microfluorimetry to measure cytoplasmic Ca2+ concentration ([Ca2+]i). The quantitative analysis of the recovery from an increase of [Ca2+]i elicited by activation of capacitative Ca2+ entry (CCE) served to characterize kinetic parameters of these Ca2+ extrusion systems in the intact cell. In CPAE cells the PMCA is activated in a Ca(2+)- and time-dependent manner. Full activation of the pump occurs only after [Ca2+]i has been elevated for at least 1 min which results in an increase of the affinity of the pump for Ca2+ and an increase in the apparent maximal extrusion rate (Vmax). Application of calmodulin antagonists W-7 and calmidazolium chloride (compound R 24571) revealed that calmodulin is a major regulator of PMCA activity in vivo. Sequential and simultaneous inhibition of PMCA and NCX suggested that both contribute to Ca2+ extrusion in a non-additive fashion. The activity of one system is dynamically adjusted to compensate for changes in the extrusion rate by the alternative transporter. It was concluded that in vascular endothelial cells, the PMCA functions as a calmodulin-regulated, high-affinity Ca2+ removal system. The contribution by the low-affinity NCX to Ca2+ clearance became apparent at [Ca2+]i > approximately 150 nM under conditions of submaximal activation of the PMCA. PMID- 10463098 TI - Mechanisms of Ca2+ store depletion in single endothelial cells in a Ca(2+)-free environment. AB - Depletion of agonist-sensitive Ca2+ stores results in activation of capacitative Ca2+ entry (CCE) in endothelial cells. The proportion of Ca2+ stores contributing to the regulation of CCE is unknown. In fura-2/am loaded single endothelial cells freshly isolated from bovine left circumflex coronary arteries, we investigated whether a resting period in a Ca(2+)-free environment results in emptying of bradykinin-sensitive Ca2+ stores (BsS) and activation of CCE. In a Ca(2+)-free environment, depletion of BsS occurred in a time-dependent manner (59% after 10 min in Ca(2+)-free solution). This effect was prevented by inhibition of the Na(+)-Ca2+ exchange but not by a blockade of ryanodine-sensitive Ca2+ release (RsCR). In contrast to BsS, mitochondrial Ca2+ content remained unchanged in the Ca(2+)-free environment. Remarkably, activity of CCE (monitored as Mn2+ influx) did not increase after depletion of BsS in the Ca(2+)-free environment. In contrast to Mn2+ influx, the effect of re-addition of Ca2+ to elevate bulk Ca2+ concentration ([Ca2+]b) decreased with the time the cells rested in Ca(2+)-free buffer. This decrease was prevented by an inhibition of RsCR. In low Na+ conditions the effect of Ca2+ on [Ca2+]b was reduced while it did not change the time the cells rested in Ca(2+)-free solution. After a 2 min period in low Na+ conditions, ryanodine-induced Ca2+ extrusion was markedly diminished. Inhibition of RsCR re-established the effect of Ca2+ on [Ca2+]b in low Na+ conditions. Collapsing subplasmalemmal Ca2+ stores with nocodazole, increased the effect of Ca2+ on [Ca2+]b. In nocodazole-treated cells, the effect of Ca2+ on [Ca2+]b was not reduced in Ca(2+)-free environment. These data indicate that activation of CCE is not associated with the agonist-sensitive Ca2+ pools that deplete rapidly in a Ca(2+)-free environment. Subplasmalemmal ryanodine-sensitive Ca2+ stores (RsS) are emptied in Ca(2+)-free/low Na+ solution and re-sequester Ca2+ which enters the cells prior an increase in [Ca2+]b occurs. Thus, in endothelial cells there are differences in the functions of various subplasmalemmal Ca2+ stores (i.e. BsS and RsS), which include either activation of CCE or regulation of subplasmalemmal Ca2+. PMID- 10463099 TI - Glucose and tolbutamide trigger transients of Ca2+ in single pancreatic beta cells exposed to tetraethylammonium. AB - Isolated pancreatic beta-cells respond to glucose stimulation with increase of the cytoplasmic Ca2+ concentration ([Ca2+]i) in terms of membrane-derived slow oscillations (0.2-0.5/min) with superimposed transient of intracellular origin. To evaluate under which conditions transients may result also from entry of extracellular Ca2+, the cytoplasmic concentration of the ion was measured with dual wavelength fluorometry and fura-2 in individual mouse beta-cells exposed to the K+ channel blocker tetraethylammonium (TEA). In the presence of 20 mM TEA, the beta-cells responded to closure of the KATP channels (increase of the glucose concentration to 11 mM or addition of 1 mM tolbutamide) with pronounced transients of [Ca2+]i. However, there were no transients when the beta-cells were depolarized by raising extracellular K+ to 30 mM in the presence of 20 mM TEA. The glucose-induced [Ca2+]i transients became more pronounced after thapsigargin inhibition of the endoplasmic reticulum Ca(2+)-ATPase. The tolbutamide-induced transients were amplified when promoting the entry of Ca2+ (rise of extracellular Ca2+ to 10 mM or addition of BAY K 8644), unaffected in the presence of thapsigargin and the Na+ channel blocker tetrodotoxin and slightly reduced by glucagon. Blockage of voltage-dependent Ca2+ channels with methoxyverapamil resulted in a prompt disappearance of the transients induced by glucose or tolbutamide. The observations indicate that closure of the KATP channels can precipitate pronounced transients of [Ca2+]i when other K+ conductances are suppressed. PMID- 10463100 TI - Impairment of myocardial calcium homeostasis by antibodies against the adenine nucleotide translocator. AB - The adenine nucleotide translocator (ANT) is an autoantigen in myocarditis and dilated cardiomyopathy. Carrier-specific antibodies impair myocardial energy metabolism and heart function. They cross-react with a myolemmal calcium channel and alter calcium fluxes in isolated myocytes. To test whether antibodies against the ANT can alter calcium homeostasis in intact hearts, guinea pigs were immunized with the carrier protein and their isolated hearts loaded with the intracellular calcium indicator INDO-1. The diastolic and systolic ratios of fluorescence signals at 410 nm and 510 nm (emission wavelengths of the calcium bound and calcium-free indicator), 'd-s410/510', were measured by excitation at 364 nm. This index of the transient calcium concentration associated with the contraction cycle correlated with the external heart work (EHW) in non-immunized controls. EHW of immunized animals was lower (76 +/- 62 vs 153 +/- 47 mJ/g/min in controls, p < 0.005) and the amplitude of d-s410/510 was elevated (27.6 +/- 4.1% of the average ratio of the whole heart cycle vs 21.7 +/- 1.2% in controls, p < 0.005) and essentially independent of EHW. Isoproterenol stimulation increased EHW in all hearts but d-s410/510 was hightened in control hearts, only. Thus, a disorder between cytosolic calcium transients and work was recorded in hearts from guinea pigs immunized with the ANT. It may contribute to an immunopathic mechanism of heart failure subsequent to myocarditis. PMID- 10463101 TI - Expression patterns of sarco/endoplasmic reticulum Ca(2+)-ATPase and inositol 1,4,5-trisphosphate receptor isoforms in vascular endothelial cells. AB - Expression patterns of sarcoplasmic/endoplasmic-reticulum Ca(2+)-ATPase (SERCA) and inositol 1,4,5-trisphosphate receptor (IP3R) isoforms were studied in endothelial cells at the mRNA level by ratio RT-PCR technique and subsequent restriction-enzyme analysis. Three types of cells have been used in the present study: rat adrenal medulla microvascular endothelial cells (RAMEC), rat aortic endothelial cells (RAEC), and human umbilical vein endothelial cells (HUVEC). Our data show the presence of multiple SERCA and IP3R isoforms in each type of endothelial cells. Freshly isolated HUVEC were an exception in this respect since they contained only SERCA3 without SERCA2b messengers. The expression patterns changed upon cell proliferation: SERCA3 and IP3R-1 messengers decreased, while IP3R-3 increased with culturing. Upon cell differentiation, induced by culturing the cells on Matrigel, the expression pattern of the IP3R changed even further in all endothelial cell types: IP3R-1 was reduced in all three cell kinds, while IP3R-3 raised significantly in RAEC and RAMEC. In HUVEC the expression of SERCA returned, upon differentiation, to the levels observed in the freshly isolated cells. Thus, the plasticity of expression of various SERCA and IP3R isoforms shows that possibly different Ca2+ pools may play distinct roles in cell proliferation and differentiation. PMID- 10463102 TI - Release of O2- by human umbilical cord blood-derived eosinophils: role of intra- and extracellular calcium. AB - The aim of our study was to investigate the physiologic mechanisms involved in eosinophil activation as an essential prerequisite to disrupting the biochemical cascade that triggers inflammation, thereby attenuating the effect of this activation or, ideally, preventing it from occurring. We have, therefore, examined the nature of the fMLP- and PAF-induced [Ca2+]i rise and the relationship between the [Ca2+]i rise and O2- production in human umbilical cord blood-derived eosinophils cultured in the presence of IL-3 and IL-5. These cells responded to fMLP or PAF (1 microM each) with an increase in [Ca2+]i (217.3 +/- 22.1 and 197.8 +/- 22.1 nM respectively) which was associated with production of O2- (40.2 +/- 8.2 and 35.2 +/- 7.6 pmol/min/10(6) cells respectively). The role of Ca2+ in the induced respiratory burst was studied by changing the availability of Ca2+ in the intra- and extracellular compartments. Removal or chelation of extracellular Ca2+ induced a reduction of both the fMLP and PAF-induced [Ca2+]i rise and O2- production. Chelation of intracellular Ca2+ induced a concentration dependent inhibition of fMLP- and PAF-induced [Ca2+]i rise and caused a decrease in O2- production. SK&F 96365 had a stimulatory effect on PAF-induced [Ca2+]i rise and on fMLP-induced O2- production, this phenomenon was not observed with extracellular Ca2+ removal or chelation. Furthermore, Ni2+ exhibited an inhibition of both fMLP and PAF-induced [Ca2+]i rise and O2- production. Finally, both fMLP and PAF induced an increase in divalent cation influx that was further augmented by thapsigargin. Our results indicate that fMLP and PAF dependent O2- production in human eosinophils require intra- and extracellular Ca2+ and that Ca2+ influx is necessary for optimal activation. PMID- 10463104 TI - Mission statement content and hospital performance in the Canadian not-for-profit health care sector. AB - This article presents the findings from an exploratory research study that assessed the content and impact of mission statements in 103 Canadian not-for profit health care organizations. The study especially sought to determine if a relationship existed between selected mission statement components and various hospital performance indicators. PMID- 10463103 TI - Cross-gender mentoring and career development in the health care industry. AB - Career development in the next century will be predicated upon the ability to forge hierarchical as well as lateral relationships. This study of 223 managers in the health care industry found that mentors influenced the career mobility of female executives and ensured access to promotions within the organization. PMID- 10463105 TI - Expanding Internet access for health care consumers. AB - Sophisticated health care consumers are beginning to use the Internet to educate themselves about their own health and manage their own care. As health care stakeholders (providers, payers, employers) feel pressure from consumers to implement Internet-related strategies, stakeholders must realize that obtaining Internet access is a challenge for many consumers. Stakeholders who expand consumer Internet access will, however, have a competitive advantage. This article outlines how stakeholders can expand consumer Internet access. PMID- 10463106 TI - Are rural hospitals "strategic"? AB - This article focuses on strategic management of rural hospitals from an evolutionary perspective, using logistic regression models to examine the environmental/organizational pressures that induced rural hospitals to be more active in strategic changes (high changers versus low changers) in two 5-year consecutive intervals between 1983 and 1993. Findings suggest that these pressures had inconsistent effects and high changers were more sensitive to organizational pressures than environmental pressures. PMID- 10463107 TI - Multiple-perspectives model of medical technology. AB - A model of medical technology is proposed, containing six different perspectives of its definition: physical, information, knowledge, process, change, and as an enabling and strategic resource. These perspectives are integrated to form an organizational dimension. The contributions of this model to better management of medical technology are described and its relation to other models in the literature is discussed. PMID- 10463110 TI - [Tobacco smoking and hearing in the elderly. Can presbycusis be prevented by smoking cessation?]. PMID- 10463108 TI - Avoidable costs of comprehensive case management. AB - Comprehensive case management has become an industry standard and its pervasiveness raises questions about the ubiquitous need for this service. Analyzed from the perspective of transaction cost analysis and access, we argue that in some cases comprehensive case management is an avoidable cost incurred because of system problems that limit access to otherwise eligible clients. Implications are discussed. PMID- 10463109 TI - Evaluations of care by ambulatory surgery patients. AB - As medical care increasingly moves from inpatient to outpatient delivery settings, attention must be paid to patient assessments of the care received in these ambulatory medical centers. This study of patients in ambulatory surgery centers (N = 275) across the United States reports on the ratings given to the varied attributes of the ambulatory surgery experience. The respondents (N = 36,078) gave highest ratings to those items which evaluated the nurses and physicians and lower marks to items related to registration and access. The article concludes with an illustration of a quadrant analysis technique that managers can use to identify the specific care issues that should be addressed to increase positive word-of-mouth referrals. PMID- 10463111 TI - [Hyposensitization in allergic rhinoconjunctivitis]. PMID- 10463112 TI - [Specific immunotherapy (hyposensitization) in allergic rhinoconjunctivitis. Meta analysis of effectiveness and side effects]. AB - BACKGROUND: Specific immunotherapy (SIT) of allergic disorders as introduced by Freeman and Noon involves the application of gradually increasing doses of extracts of the material to which the individual is sensitive. SIT represents the only specific treatment that can be offered to allergic patients besides allergen avoidance. SIT has been widely used in patients with pollen allergic rhinitis and its clinical efficacy has been demonstrated in several controlled clinical trials. The underlying mechanism of this treatment is still not understood. Previous studies have focused on changes in serum antibodies, including blunting of seasonal rises in specific IgE and increases in specific IgG antibodies, especially of the IgG4 isotype. Recent studies have suggested an effect on T lymphocytes, leading to a switch from a predominant Th2 response--that is, interleukin (IL)-4 and IL-5--to a Th1 response involving interferon (IFN)-gamma. The switch of a cytokine profile to a predominant IFN-gamma response results in inhibition of IL-4 dependent IgE production, which is reinforced by a decrease in the production of IL-4 by Th2 cells. METHODS: In the present study a meta analysis was performed to evaluate all double-blind placebo-controlled studies investigating the effect of SIT on clinical and immunological treatment used in patients with allergic rhinoconjunctivitis. RESULTS: SIT was shown to be an effective treatment modality in allergic rhinoconjunctivitis, decreasing symptoms, the need for medication and reactivity in specific nasal and conjunctival provocation tests, as well as inflammatory markers. Histologically, a switch might be induced from a predominant Th2 cell profile to that of a Th1. CONCLUSIONS: In general, the efficacy of SIT is dependent on the allergen the individual patient is sensitive to, the quality and total amount of the allergen applied and the SIT schedule. PMID- 10463113 TI - [Surgery of acquired laryngotracheal stenoses in childhood. Experiences and results from 1988-1998. II: The cricotracheal resection]. AB - Approximately 90% of infants and children with severe acquired laryngotracheal stenoses are tracheotomy dependent and therefore impaired in their physical and speech developments. In addition, tracheotomized infants can be endangered by the cannula due to the possible crusting of secretions or its dislocation. Thus, early repair of a stenosis is mandatory. Within the last 10 years, we successfully operated on 18 children with severe laryngotracheal stenoses. Ten children were treated with a modified Cotton technique. This paper reports our results of cricotracheal resection performed in 8 children since 1994 (age distribution: 7 months through age 15 years). Four children had Cotton grade II stenoses, three had grade III stenoses and one grade IV stenoses. In 3 patients a tracheotomy had been performed at another institution. Since their tracheostomas were too far caudal, they could not be included in the primary resection. All 8 children have been successfully decannulated. Five children without tracheotomies could be extubated uneventfully on the 5th postoperative day. All three primarily tracheotomized children needed further endotracheal stenting with T-tubes because of stomal and suprastomal collapse. Two of these latter children additionally required a tracheoplasty with rib cartilage grafts in order to stabilize the suprastomal trachea prior to decannulation. No patient experienced injuries to the recurrent laryngeal nerves or insufficiencies of the anastomosis. All children's voices were not impaired. This is the third report in literature of cricotracheal resections in infants and children, indicating that this effective, one-stage procedure is superior to laryngotracheal reconstruction with rib cartilage. PMID- 10463114 TI - [Prosthetic management of tumor-induced nasal defects]. AB - Reconstructive plastic surgery is not always possible or advisable for nasal defects following tumor resections. Facial prostheses offer an alternative therapy. Since 1991, 37 patients with nasal defects after tumor surgery were treated in the Berlin Center of Facial Prostheses: 28 received "soft" silicone facial prostheses and 9 "solid" polymethylmetacrylate facial prostheses. In 35 patients protheses were bone-anchored: Branemark implants were used in 12 patients and an Epitec frame construction in 23 patients. Glue-on prostheses were applied to only two patients. No intraoperative complications occurred at implant insertion. During following visits only minor inflammations were observed around implants and were treated locally. The implant success rate was 78% for the Branemark implants (89.7% in non-radiated areas and 50% in radiated areas) and 100% in radiated and non-radiated areas for the Epitec system. Our investigation shows that given the right indication, the bone-anchored facial prosthesis is safe, cosmetically acceptable, uncomplicated and not stressful for the patient. PMID- 10463115 TI - [Sniffin'Sticks: a new system for olfactory assessment in routine clinical practice]. AB - INTRODUCTION: The Sniffin'Sticks test battery consists of a short screening test and tests for odor detection thresholds (using n-butanol), odor discrimination and odor identification. We evaluated the usefulness of this new test in clinical practice and propose normative values. MATERIAL AND METHODS: In the present study 30 normosmic and 15 anosmic volunteers as well as 35 patients with various olfactory dysfunctions were examined. RESULTS: All four Sniffin'Sticks tests were able to separate normosmics and anosmics in a highly significant manner. Even a small decrease in olfactory performance was detectable in elderly patients. For practical purposes the sensitivity of the test for screening test was found to be too low. The good correlation between the individual Sniffin'Sticks tests and the correlation between the Sniffin'Sticks test battery and the "University of Pennsylvania Smell Identification Test" (UPSIT) test battery and the "University of Pennsylvania Smell Identification Test" (UPSIT) document the reliability of these tests. CONCLUSIONS: Our findings show that the Sniffin'Sticks are well suited to assess olfactory performance in clinical practice. PMID- 10463116 TI - [Sjogren syndrome and bilateral MALT lymphoma of the parotid gland]. AB - Sjogren's syndrome is an important autoimmune disease in the head and neck. Patients have an increased arrival risk of up to 6% per year for developing B cell lymphomas, including mucosa-associated lymphoid tissue (MALT) lymphomas. The following case report shows this relation and the difficulty of differentiating clinically recurrent swelling of the parotid gland in Sjogren's syndrome from malignant lymphoma. A 64-year-old woman had a 2-year history of indolent, recurrent swelling of both parotid glands. Blood examination showed elevated ESR and a hypergammaglobulinemia. Immunosuppressive therapy produced no improvement. Two years after the diagnosis of Sjogren's syndrome, swelling of the left parotid gland persisted. Superficial parotidectomy of the left side was performed and histopathological examination revealed a MALT-related lymphoma. Subsequent parotidectomy of the right side also showed infiltration of the gland by a MALT lymphoma. Postoperative radiation therapy was given. During the follow-up period no recurrence or systemic disease was detected. Patients with Sjogren's syndrome should be examined regularly by the otolaryngologist. If a lymphoma cannot be ruled out, open biopsy must be considered for histological diagnosis. Prognostic factors for developing a lymphoma are possibly a high ESR and hypergammaglobulinemia. Further prognostic factors have to be evaluated. PMID- 10463117 TI - [Cochlear prosthesis implantation in superficial leptomeningeal hemosiderosis]. AB - Superficial leptomeningeal hemosiderosis is a rare disease of the central nervous system. Chronic bleeding into the subarachnoid space causes deposition of hemosiderin in glial cells and subsequent damage to adjacent brain tissue. There is a characteristic predilection for the cerebellum and eighth cranial nerve. Accordingly, among a variety of symptoms, cerebellar ataxia and sensorineural hearing loss progressing to total deafness commonly occur. To date, the hearing loss has been believed to be purely neural. We present a case of superficial hemosiderosis in a patient with total deafness who was successfully provided with a cochlear implant. Audiometry demonstrated total bilateral cochlear hearing losses but with preserved function of the eighth cranial nerve. These findings make us conclude that in contrast to current opinion, superficial hemosiderosis can initially damage the cochlea alone. Thus, patients with total deafness due to this disorder may benefit from cochlear implantation. All patients require careful audiometric assessment, including promontory stimulation. PMID- 10463118 TI - [Iatrogenic Cushing syndrome and mutatio tarda caused by dexamethasone containing nose drops]. AB - Nasal steroid drops can produce iatrogenic Cushing's syndrome if the recommended dose is exceeded. We now report a 19-year-old male patient for whom dexamethasone nasal drops were prescribed because of nasal obstruction. He had been using these drops for more than 5 years, with a daily dexamethasone dose of 0.7-1.0 mg. It is likely that significant amounts of dexamethasone were swallowed and absorbed by the patient's gastrointestinal tract. As a consequence, the patient developed Cushing's syndrome with panhypopituitarism, growth retardation, osteoporosis and hypertension. A boyish size of the larynx resulted in a fundamental speaking frequency of a child. After discontinuing the intranasal use of the of dexamethasone and adjusting doses of hydrocortisone over 3.5 months a normalization of endocrine function occurred with the beginning of puberty and normal sexual changes. These findings show that the use of nasal steroid drops particularly in children should be limited to a short period and drugs with a low absorption dose employed. PMID- 10463119 TI - [Treatment refractory unilateral headache. Single focus Langerhans cell histiocytosis (eosinophilic granuloma) of the pterygopalatine fossa]. PMID- 10463120 TI - ["Therapy refractory" exophthalmos. Orbital hemangioma]. PMID- 10463121 TI - [Histologic examination of surgically removed body tissue]. PMID- 10463122 TI - [Drug costs in patients with chronic complex tinnitus]. PMID- 10463123 TI - [Recommendations for medical documentation in suspected occupational hearing loss]. PMID- 10463124 TI - [Increasing the length of the columella. Concepts and techniques]. PMID- 10463125 TI - Nonlinear multiscale analysis of three-dimensional echocardiographic sequences. AB - We introduce a new model for multiscale analysis of space-time echocardiographic sequences. The proposed nonlinear partial differential equation, representing the multiscale analysis, filters the sequence while keeping the space-time coherent structures. It combines the ideas of regularized Perona-Malik anisotropic diffusion and the Galilean invariant movie multiscale analysis of Alvarez, Guichard, Lions and Morel. A numerical method for solving the proposed partial differential equation is suggested and its stability is shown. Computational results on synthesized and real sequences are provided. A qualitative and quantitative evaluation of the accuracy of the method is presented. PMID- 10463126 TI - Reconstruction of the human cerebral cortex from magnetic resonance images. AB - Reconstructing the geometry of the human cerebral cortex from MR images is an important step in both brain mapping and surgical path planning applications. Difficulties with imaging noise, partial volume averaging, image intensity inhomogeneities, convoluted cortical structures, and the requirement to preserve anatomical topology make the development of accurate automated algorithms particularly challenging. In this paper we address each of these problems and describe a systematic method for obtaining a surface representation of the geometric central layer of the human cerebral cortex. Using fuzzy segmentation, an isosurface algorithm, and a deformable surface model, the method reconstructs the entire cortex with the correct topology, including deep convoluted sulci and gyri. The method is largely automated and its results are robust to imaging noise, partial volume averaging, and image intensity inhomogeneities. The performance of this method is demonstrated, both qualitatively and quantitatively, and the results of its application to six subjects and one simulated MR brain volume are presented. PMID- 10463127 TI - Off-resonance correction of MR images. AB - In magnetic resonance imaging (MRI), the spatial inhomogeneity of the static magnetic field can cause degraded images if the reconstruction is based on inverse Fourier transformation. This paper presents and discusses a range of fast reconstruction algorithms that attempt to avoid such degradation by taking the field inhomogeneity into account. Some of these algorithms are new, others are modified versions of known algorithms. Speed and accuracy of all these algorithms are demonstrated using spiral MRI. PMID- 10463128 TI - Nonactive antenna compensation for fixed-array microwave imaging--Part I: Model development. AB - Fixed-array microwave imaging with multisensor data acquisition can suffer from nonactive antenna element interactions which cause distortions in the measurements. In Part I of a two-part paper, we develop a nonactive antenna compensation model for incorporation in model-based near-field microwave image reconstruction methods. The model treats the nonactive members of the antenna array as impedance boundary conditions applied over a cylindrical surface of finite radius providing two parameters, the effective antenna radius and impedance factor, which can be determined empirically from measured data. Results show that the effective radius and impedance factor provide improved fits to experimental data in homogeneous phantoms where measurements are obtained with and without the presence of the nonactive antenna elements. Once deduced, these parameters are incorporated into the nonactive antenna compensation model and lead to systematic data-model match improvements in heterogeneous phantoms. While the improvements afforded by the nonactive antenna model are small on a per measurement basis, they are not insignificant. As shown in Part II, inclusion of this new model for nonactive antenna compensation produces significantly higher quality image reconstructions from measurements obtained with a fixed-array data acquisition system over the frequency band 500-900 MHz. PMID- 10463129 TI - Nonactive antenna compensation for fixed-array microwave imaging: Part II- Imaging results. AB - Model-based imaging techniques utilizing microwave signal illumination rely heavily on the ability to accurately represent the wave propagation with a suitable numerical model. To date, the highest quality images from our prototype system have been achieved utilizing a single transmitter/single receiver measurement system where both antennas are manually repositioned to facilitate multiple illuminations of the imaging region, thus requiring long data acquisition times. In an effort to develop a system that can acquire data in a real time manner, a 32-channel network has been fabricated with all ports capable of being electronically selected for either transmit or receive mode. The presence of a complete array of antenna elements at data collection time perturbs the field distributions being measured, which can subsequently degrade the image reconstruction due to increased data-model mismatch. Incorporating the nonactive antenna-compensation model from Part I of this paper into our hybrid element near field image reconstruction algorithm is shown to restore image quality when fixed antenna-array data acquisition is used. Improvements are most dramatic for inclusions located in near proximity to the antenna array itself, although cases of improvement in the recovery of centered heterogeneities are also illustrated. Increases in the frequency of illumination are found to warrant an increased need for nonactive antenna compensation. Quantitative measures of recovered inclusion shape and position reveal a systematic improvement in image reconstruction quality when the nonactive antenna-compensation model is employed. Improvements in electrical property value recovery of localized heterogeneities are also observed. Image reconstructions in freshly excised breast tissue illustrate the applicability of the approach when used with our two-dimensional microwave imaging system. PMID- 10463130 TI - Anti-aliased three-dimensional cone-beam reconstruction of low-contrast objects with algebraic methods. AB - This paper examines the use of the algebraic reconstruction technique (ART) and related techniques to reconstruct 3-D objects from a relatively sparse set of cone-beam projections. Although ART has been widely used for cone-beam reconstruction of high-contrast objects, e.g., in computed angiography, the work presented here explores the more challenging low-contrast case which represents a little-investigated scenario for ART. Preliminary experiments indicate that for cone angles greater than 20 degrees, traditional ART produces reconstructions with strong aliasing artifacts. These artifacts are in addition to the usual off midplane inaccuracies of cone-beam tomography with planar orbits. We find that the source of these artifacts is the nonuniform reconstruction grid sampling and correction by the cone-beam rays during the ART projection-backprojection procedure. A new method to compute the weights of the reconstruction matrix is devised, which replaces the usual constant-size interpolation filter by one whose size and amplitude is dependent on the source-voxel distance. This enables the generation of reconstructions free of cone-beam aliasing artifacts, at only little extra cost. An alternative analysis reveals that simultaneous ART (SART) also produces reconstructions without aliasing artifacts, however, at greater computational cost. Finally, we thoroughly investigate the influence of various ART parameters, such as volume initialization, relaxation coefficient lambda, correction scheme, number of iterations, and noise in the projection data on reconstruction quality. We find that ART typically requires only three iterations to render satisfactory reconstruction results. PMID- 10463131 TI - Fast implementations of algebraic methods for three-dimensional reconstruction from cone-beam data. AB - The prime motivation of this work is to devise techniques that make the algebraic reconstruction technique (ART) and related methods more efficient for routine clinical use, while not compromising their accuracy. Since most of the computational effort of ART is spent for projection/backprojection operations, we first seek to optimize the projection algorithm. Existing projection algorithms are surveyed and it is found that these algorithms either lack accuracy or speed, or are not suitable for cone-beam reconstruction. We hence devise a new and more accurate extension to the splatting algorithm, a well-known voxel-driven projection method. We also describe a new three-dimensional (3-D) ray-driven projector that is considerably faster than the voxel-driven projector and, at the same time, more accurate and perfectly suited for the demands of cone beam. We then devise caching schemes for both ART and simultaneous ART (SART), which minimize the number of redundant computations for projection and backprojection and, at the same time, are very memory conscious. We find that with caching, the cost for an ART projection/backprojection operation can be reduced to the equivalent cost of 1.12 projections. We also find that SART, due to its image based volume correction scheme, is considerably harder to accelerate with caching. Implementations of the algorithms yield run-time ratios TSART/TART between 1.5 and 1.15, depending on the amount of caching used. PMID- 10463133 TI - Comments on "Retrospective motion correction in digital subtraction angiography: a review". PMID- 10463132 TI - Adaptive template filtering for signal-to-noise ratio enhancement in magnetic resonance imaging. AB - In this paper, a local shape-adaptive template filtering is proposed for the enhancement of the signal-to-noise ratio (SNR) without the loss of resolution in magnetic resonance (MR) imaging. Unlike conventional filtering, where the template shape and coefficients are fixed, multiple templates are defined in the proposed algorithm. An optimal template is selected and optimal filtering, based on the template, is applied on a pixel-by-pixel basis. Using the proposed process, edge blurring is minimized and SNR enhancement is maximized by selecting the optimally matched template. Compared to existing two-dimensional (2-D) adaptive linear least square error (LLSE) filters or direction-adaptive recursive filters, the proposed adaptive template filter provides higher SNR and sharper edges for both MR and artificial resolution phantom images. PMID- 10463134 TI - Attenuation by acetyl-L-carnitine of neurological damage and biochemical derangement following brain ischemia and reperfusion. AB - Alterations in brain metabolism after ischemia and reperfusion are described herein. Several roles played by carnitine and acetylcarnitine can be of particular relevance in counteracting these brain metabolism alterations. The effects of acetylcarnitine in several experimental models of brain ischemia in rats are described. The data obtained show that acetylcarnitine can have significant clinical neuroprotective effects when administered shortly after the onset of focal or global cerebral ischemia. In the canine cardiac arrest model, acetylcarnitine improved the postischemic neurological outcome and tissue levels of lactate and pyruvate were normalized. A trend toward reversal of pyruvate dehydrogenase inhibition in acetylcarnitine-treated dogs was also observed. The immediate postischemic administration of acetylcarnitine prevents free radical mediated protein oxidation in the frontal cortex of dogs submitted to cardiac arrest and resuscitation. The transfer of the acetyl group to coenzyme A (CoA) to form acetyl-CoA as the primary source of energy is a plausible mechanism of action of acetylcarnitine. PMID- 10463135 TI - Cyclosporine-induced lipid peroxidation and propionyl carnitine protective effect. AB - Cell and tissue lipoperoxidation of the kidney induced by cyclosporine through the release of reactive oxygen species has recently been pointed out to be one of the factors responsible for the toxic phenomena related to the administration of cyclosporine. Our previous research on propionyl carnitine had shown an antilipoperoxidative effect of this substance on isolated cells such as erythrocytes and leukocytes, and also on the endothelial, vasal and cardiac tissues. In the experiments presented herein we also examined if propionyl carnitine could carry out its already well-known antilipoperoxidative effect in the renal tissue, and if this mechanism could be taken into consideration in order to explain the protective effect of propionyl carnitine against cyclosporine induced toxicity. Trials were carried out on isolated and perfused rat kidneys, and we were able to observe that propionyl carnitine exerted a protective action on toxic lipid peroxidation phenomena induced by cyclosporine. The results we obtained, together with other mechanisms which we had already proved regarding the intense protective activity of propionyl carnitine on cyclosporine-induced nephrotoxicity, complete the complex picture that describes the protective activity of propionyl carnitine against cyclosporine toxicity. PMID- 10463136 TI - Propinox in biliary colic: a multicenter, randomized, prospective and parallel double-blind study of three doses of propinox versus placebo in acute biliary colic pain. AB - The aim of this study was to assess the efficacy and tolerance of propinox administered i.v., and establish a dose-response relation according to three dose levels (10, 20 and 30 mg), vs. placebo in patients with moderate to severe acute biliary pain. Three hundred and fifty patients were included: 85 received placebo treatment, 81 were treated with propinox 10 mg, 91 with propinox 20 mg and 93 received propinox 30 mg. Spontaneous pain intensity was assessed according to a visual analog and a verbal scale before treatment and 20, 60 and 120 min after. All treatments induced significant and progressive pain reduction at all controls, but patients treated with 20 and 30 mg of propinox showed significantly lower pain intensity after 120 min compared to the placebo group. The last control revealed that 28% of patients receiving placebo had no pain while 60% of patients treated with propinox 30 mg reported absence of pain with a statistically significant difference (p < 0.001). All treatments were very well tolerated and there were no dropouts due to adverse events. Mouth dryness was the adverse effect occurring with a significantly higher frequency than that observed with placebo although it was only seen in patients treated with 20 mg and 30 mg active doses. The results of this study showed that propinox was an effective drug in the treatment of moderate to severe colic pain of biliary origin. Concerning efficacy and side effects, a clear dose-response relation was observed; the 20 mg and 30 mg doses being significantly superior to placebo. PMID- 10463137 TI - Sonographic assessment of soft tissue alterations in osteoarthritis of the knee. AB - Osteoarthritis is the most common joint disorder to affect the knee and the single most important cause of disability in elderly people. Radiological criteria for osteoarthritis of the knee (focal loss of cartilage, osteophytosis, subchondral cysts and subchondral sclerosis) reflect late manifestations and significant joint damage only. Ultrasound examination is useful in revealing disruption of the periarticular tissue, which contributes to symptoms and disability in advanced disease. Nevertheless, ultrasound examination is very important in early osteoarthritis of the knee when it provides information about soft tissues and cartilage involvement in the absence of clinical symptoms. PMID- 10463138 TI - Influence of aetiology on the effects of treatment in heart failure. PMID- 10463139 TI - The influence of beta-adrenergic blocking drugs on morbidity and mortality in heart failure. PMID- 10463140 TI - Left ventricular remodelling during treatment with beta-blockers. PMID- 10463141 TI - Effects of beta-blockers on symptoms and exercise tolerance in patients with heart failure. PMID- 10463142 TI - Acute myocardial infarction: psychosocial and cardiovascular risk factors in men. AB - BACKGROUND: Controversy still exists over the independent role of psychosocial factors in acute myocardial infarction (AMI). This study aimed to compare psychometric indices in Portuguese patients surviving a first episode of AMI and a community sample free of AMI. DESIGN: A community-based, case-control study. The study sample included 153 male patients with a first episode of AMI and 156 male controls randomly selected from the community. METHODS: Information on demographic, social, medical and behavioural characteristics was obtained using a structured questionnaire; anthropometric, blood pressure, and serum measurements were performed, and the psychometric evaluation was obtained using a self evaluation questionnaires (Bortner scale, Beck Depression Inventory, SCL-90-R, Psychological General Well-Being and the Nottingham Health Profile). RESULTS: Cases of AMI more often exhibited type A behaviour (15.4 +/- 3.0 versus 14.7 +/- 2.7, P = 0.041), had more depressive symptoms (10.8 +/- 8.0 versus 8.6 +/- 8.0, P = 0.005) and showed worse scores on general well-being (68.4 +/- 17.7 versus 76.6 +/- 16.2, P < 0.001) than controls. Cases were significantly different from controls in terms of years of education (6.7 +/- 4.4 versus 9.0 +/- 4.7 years, P < 0.001), hypertension (43.1% versus 30.1%, P = 0.024), dislipidaemia (43.8% versus 34.0%, P = 0.038), type I obesity (53.6% versus 42.3%, P = 0.045), smoking (54.1% versus 26.5%, P < 0.001), no practise of exercise (68.5% versus 50.3%, P = 0.002) and presence of non-insulin-dependent diabetics (16.3% versus 6.4%, P = 0.010). After adjusting for such factors a similar set of significant psychometric results was found. CONCLUSION: Type A behaviour, depression and lower levels of well-being and quality of life, independent of other cardiovascular risk factors, were significant features of AMI cases. PMID- 10463143 TI - Ten-year cardiovascular risk in a Hong Kong population. AB - BACKGROUND: Cardiovascular disease is a major health issue in Hong Kong. We conducted a screening program to assess the 10-year risk for the population and to assess the potential benefit of large-scale screening in Hong Kong. METHOD: A local screening program for cardiovascular risk was carried out in a health service network with a total of 17,716 participants. Retrospective data analysis for the prevalence and distribution of the various risk factors was performed. The data were then applied to calculate the 10-year risk of each individual, according to the European Task Force coronary risk chart. RESULTS: Of the participants, 54.2% had total cholesterol levels > 5.2 mmol/l; 28.7% had body mass index > 25 kg/m2; 18.5% were hypertensive; 15.1% were smokers; and 3.7% had diabetes mellitus. There were 35.5% of the screened population who had at least two risk factors and 10.9% had at least three risk factors. A total of 9049 individuals satisfied the criteria for the European Task Force guidelines and were selected for 10-year cardiovascular risk analysis. We calculated that 68.0% of the male population had at least 10% risk and 41.5% had at least 20% risk of developing a coronary heart event within 10 years. Among women, 48.2% of the population carried at least 10% risk and 2.8% carried at least 20% risk. CONCLUSION: The calculated 10-year risk of the population, particularly for men, is significant. Our study demonstrated that mass screening is feasible, and has the benefit of early identification of high-risk individuals, which could be a reasonable strategy for cost-effective medicine. PMID- 10463144 TI - Lipids, lipoproteins and apolipoprotein (a) isoforms in type 2 diabetic patients. AB - BACKGROUND: Patients with type 2 diabetes mellitus have a greater than normal risk of developing atherosclerotic vascular diseases. Higher than normal plasma concentrations of lipoprotein (a) [Lp(a)] have been associated with premature atherosclerosis in several studies. OBJECTIVE: To determine the concentrations of lipids, lipoproteins, and Lp(a) in 107 type 2 diabetic patients, and the distribution of apolipoprotein (a) [apo(a)] phenotypes for this group, and to compare the results found with results for healthy subjects. RESULTS: Plasma concentrations of cholesterol, triglycerides, and apolipoprotein B in the diabetics were significantly higher than those in control subjects. Diabetic patients had slightly lower Lp(a) concentrations than did nondiabetic subjects, but these differences were not statistically significant. Distributions of Lp(a) concentrations both in type 2 diabetic patients and in control subjects were markedly skewed, the highest prevalences being of low values. CONCLUSION: Distributions of apo(a) phenotypes for patients with type 2 diabetes mellitus and controls were remarkably alike. Smaller isoforms were similarly prevalent for the two populations, as were the null, single-band and double-band apo(a) phenotypes. PMID- 10463145 TI - Prevalence of hypertension in a Mexican population according to the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure. AB - BACKGROUND: Hypertension in Mexico represents a challenging public health problem. The National Survey on Chronic Diseases published in 1993 reported that hypertension affects more than 10 million Mexicans. No information has been published regarding the prevalence of hypertension in Mexico using the new diagnostic criteria established by the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure (JNC VI). METHODS: The Mexico City Diabetes Study is a prospective study designed to estimate the prevalence and incidence of cardiovascular risk factors in a low-income area. The survey included 941 men and 1341 non-pregnant women aged 35-64 years. Blood pressure measurements were performed using a random zero sphygmomanometer. The diagnostic criteria for hypertension were those recommended by the JNC VI. RESULTS: The crude prevalence of hypertension was 17.2% and 18.1% in men and women, respectively. We found significant associations between hypertension and obesity, body fat distribution, very-low-density lipoprotein cholesterol, fasting and 2-h post-glucose in both sexes, and between hypertension and total cholesterol, low-density lipoprotein cholesterol and triglycerides levels in women. In 40% of hypertensive men and 23% of women, hypertension was undiagnosed and untreated. Of the previously diagnosed hypertensive individuals, 38% of men and 30% of women reported not taking antihypertensive medicine. The prevalence++ of associated risk factors in this population is 12.3% for tobacco consumption, 22.4% for diabetes, 49.8% for hypertriglyceridemia and 40.9% for hypercholesterolemia. CONCLUSIONS: Hypertension occurs in 18% of this population. There is a high prevalence of undiagnosed and untreated cases. Associated cardiovascular risk factors are highly prevalent. PMID- 10463146 TI - A comparison of lifestyle, genetic, bioclinical and biochemical variables of offspring with and without family histories of premature coronary heart disease: the experience of the European Atherosclerosis Research Studies. AB - BACKGROUND: Lifestyle, genetic, bioclinical and biochemical factors of European university students aged 18-26 years, with and without documented paternal histories of premature coronary heart disease, have been compared in the European Atherosclerosis Research Studies (EARS) I and II. OBJECTIVE: To highlight consistencies and inconsistencies between findings in the two studies. METHODS: All measurements were made according to strict protocols, by trained technicians using validated methods. The results for men in EARS I are compared with those from EARS II which was confined to men. RESULTS: In both studies we found no differences between cases and controls in lifestyle factors and bioclinical factors except that controls were taller. We found inconsistent differences between obesity indices and antecedents of arterial hypertension. In both studies we found consistent differences between cases and controls in levels of total cholesterol and apolipoprotein B, both levels being higher in cases. The lack of any difference between levels of high-density lipoprotein cholesterol and apolipoprotein A1 was also found consistently. Inconsistent differences were found for levels of triglycerides and apolipoprotein E. For most of the candidate genes that were studied, no differences between cases and controls were found, but different polymorphisms were associated with levels of lipids, apoproteins and fibrinogen independently of case-control status. Some of these associations were potentiated by lifestyle factors. The interaction between genetic and environmental factors is further illustrated with results from the association of apolipoprotein E polymorphism with the level of apolipoprotein B and a variety of other determinants of apolipoprotein B level. CONCLUSIONS: In the EARS studies a documented family history of premature coronary heart disease was mainly expressed in terms of biochemical factors that are determined both by nature and by nurture. PMID- 10463147 TI - Bibliography. Current world literature. PMID- 10463148 TI - Microbial ubiquinones: multiple roles in respiration, gene regulation and oxidative stress management. PMID- 10463149 TI - Growth inhibition of Clostridium cellulolyticum by an inefficiently regulated carbon flow. AB - Carbon flow in Clostridium cellulolyticum was investigated either in batch or continuous culture using a synthetic medium with cellobiose as the sole source of carbon and energy. Previous experiments carried out using a complex growth medium led to the conclusion that the carbon flow was stopped by intracellular NADH. In this study, results showed that cells cultured in a synthetic medium were better able to control electron flow since the NADH/NAD+ ratios were in the range 0.3 0.7, whereas a ratio as high as 57 was previously found in cells cultured on a complex medium. Furthermore, a specific rate of cellobiose consumption of 2.13 mmol (g cells)-1 h-1 was observed on synthetic medium whereas the highest value obtained on complex medium was 0.68 mmol (g cells)-1 h-1. When C. cellulolyticum was grown in continuous culture and cellobiose in the feed medium was increased from 5.84 to 17.57 mM in stepwise fashion, there was an increase in cellobiose utilization without growth inhibition. In contrast, when the reactor was fed directly with 14.62 mM cellobiose, residual cellobiose was observed (4.24 mM) and growth was limited. These data indicate that C. cellulolyticum is not able to optimize its growth and carbon flow in response to a sudden increase in the concentration of growth substrate cellobiose. This interpretation was confirmed (i) by the study of cellobiose batch fermentation where it was demonstrated that growth inhibition was not due to nutritional limitation or inhibition by fermentation products but was associated with carbon excess and (ii) by the growth of C. cellulolyticum in dialysis culture where no growth inhibition was observed due to the limitation of carbon flow by the low rate of cellobiose diffusion through the dialysis tubing. PMID- 10463150 TI - The glyceraldehyde-3-phosphate dehydrogenase of Clostridium acetobutylicum: isolation and purification of the enzyme, and sequencing and localization of the gap gene within a cluster of other glycolytic genes. AB - Glyceraldehyde-3-phosphate dehydrogenase was purified from Clostridium acetobutylicum by sequential ammonium sulfate precipitation, gel filtration and anion-exchange chromatography (to a specific activity of 27 U mg-1). The enzyme had a molecular mass of 40 kDa as determined by SDS-PAGE and a native molecular mass of 160 kDa as determined by nondenaturing PAGE, indicating that it has a homotetrameric composition. Its pH optimum was between 8.5 and 9.3. The corresponding gene (gap) was cloned and sequenced from C. acetobutylicum DSM 792 and found to cluster with other genes of enzymes from the glycolytic pathway (pgk, phosphoglycerate kinase; tpi, triosephosphate isomerase; pgm(i), 2,3 bisphosphoglycerate-independent phosphoglycerate mutase). No sequences resembling rho-independent transcription terminators were found in the intergenic regions. A plasmid carrying the clostridial gap gene complemented an Escherichia coli gap mutant. PMID- 10463151 TI - Regulated interactions between partner and non-partner sensors and response regulators that control glycopeptide resistance gene expression in enterococci. AB - Transcription of the vanA and vanB glycopeptide resistance gene clusters is regulated by the VanRS and VanRBSB two-component regulatory systems, respectively. Histidine to glutamine substitutions were introduced at positions 164 of VanS and 233 of VanSB to prevent autophosphorylation of the sensor kinases and transfer of the phosphate groups to the VanR and VanRB response regulators. VanSH164Q and VanSBH233Q abolished activation of VanR and VanRB by host kinases. The phosphatase activity of VanSBH233Q was negatively modulated by vancomycin whereas VanSH164Q prevented transcription of the resistance genes under all growth conditions. Cross-talk was detected between VanRB and VanS in a vanSB null mutant. VanR is required for activation of promoters PR and PH allowing transcription of the regulatory (vanRS) and resistance (vanHAXYZ) genes, respectively. Under non-inducing conditions, activation of VanR by cross-talk was blocked by the presence of a multicopy plasmid carrying PH. Presence of the high affinity VanR-binding sites of the regulatory region of PH on the multicopy vector probably sequestered VanR, thereby preventing autoactivation of the PR promoter. Under such circumstances, stimulation of the host kinase by glycopeptides or moenomycin was required for expression of the resistance genes. PMID- 10463152 TI - Genetic and physiological studies of the CiaH-CiaR two-component signal transducing system involved in cefotaxime resistance and competence of Streptococcus pneumoniae. AB - A mutation in the ciaH gene of Streptococcus pneumoniae induces cefotaxime resistance and transformation deficiency. ciaH encodes a putative sensor protein that belongs to the family of signal-transducing histidine kinases. This gene is adjacent to ciaR, which encodes a DNA-binding protein involved in the regulation of genes responding to environmental signals sensed by the histidine kinase. The authors have characterized a mutation that induces reversion of both cefotaxime resistance and transformation deficiency. It is a T/A deletion in the ciaR gene resulting in the synthesis of a truncated protein containing only 125 amino acids instead of 224. The ciaH mutation requires a functional CiaR protein for expression. Northern blot analysis, using ciaR-ciaH as a probe, revealed one mRNA from the wild-type strain, indicating that the two genes constitute an operon. Comparisons of Northern blots show that the operon is constitutively activated in the strain carrying only the ciaH mutation. In the wild-type strain the activation occurs when the Ca2+ concentration is very low, demonstrating that Ca2+ is the environmental signal. The pleiotropic effects caused by the ciaH mutation include sensitivity to antibiotics and toxins, the ability to form protoplasts and the susceptibility to lysis with deoxycholate. Null-mutants were constructed in both genes and the particular features of the ciaR null mutant determined. It is able to grow in choline-deprived medium, and competence development occurs in a phosphate-deprived competence medium (CH-maleate), suggesting that the CiaH-CiaR system regulates several pathways, including teiochoic acid synthesis. PMID- 10463153 TI - Phylogenetic analysis and rapid identification of Candida dubliniensis based on analysis of ACT1 intron and exon sequences. AB - The phylogenetic position of Candida dubliniensis has previously been established on the basis of the sequence of rRNA genes. In order to confirm the relationship between C. dubliniensis and other yeast species, particularly Candida albicans, using non-rRNA gene sequences the ACT1 gene was chosen for analysis. Three overlapping fragments that together span the entire C. dubliniensis ACT1 gene (CdACT1) were amplified from a recombinant phage isolated from a genomic DNA lambda library using PCR. These were cloned and used to determine the contiguous sequence of the gene. Analysis of the sequence data revealed the presence of a 1131 bp ORF interrupted by a single 632 bp intron at the 5' extremity of the gene. Comparison of the CdACT1 sequence with the C. albicans homologue (CaACT1) revealed that although the exons are 97.9% identical the introns are only 83.4% identical. Phylogenetic trees generated using ACT1 exon and intron sequences from a range of yeast species unequivocally confirmed the phylogenetic position of C. dubliniensis as a unique taxon within the genus Candida. Analysis of the ACT1 associated intron sequences from 10 epidemiologically unrelated C. dubliniensis isolates from disparate geographical locations showed a very low level of intraspecies sequence variation. In order to develop an accurate and rapid method to identify C. dubliniensis from primary isolation plates the significant divergence between the C. dubliniensis and C. albicans ACT1 intron sequences was exploited by designing C. dubliniensis-specific PCR primers. Using a rapid boiling method to produce template DNA directly from colonies from primary isolation plates in 10 min, these primers were used in a blind test with 122 isolates of C. dubliniensis, 53 isolates of C. albicans, 10 isolates of C. stellatoidea and representative isolates of other clinically relevant Candida and other yeast species. Only the C. dubliniensis isolates yielded the C. dubliniensis-specific 288 bp amplimer. Use of this technique on colonies suspected to be C. dubliniensis allows their correct identification as C. dubliniensis in as little as 4 h. PMID- 10463154 TI - High cAMP in spores of Dictyostelium discoideum: association with spore dormancy and inhibition of germination. AB - Signalling mechanisms involving cAMP have a well-documented role in the coordination of multicellular development and differentiation leading to spore formation in the social amoeba, Dictyostelium discoideum. The involvement of cAMP in the poorly understood developmental stages of spore dormancy and germination have been investigated in this study. Dormant spores contained up to 11-fold more cAMP than nascent amoebae. The spore cAMP levels were not constant, but typically underwent a surge at 14-18 d when spores acquired the ability to germinate spontaneously. The high cAMP levels decreased only during successful spore germination, i.e. emergence of nascent amoebae. The temporal pattern of cAMP decrease was complex and unique to the method of spore activation, supporting our hypothesis that exogenously (e.g. heat) activated and autoactivated spores germinate by different mechanisms. During heat-induced activation, transcription of acg (a gene encoding adenylyl cyclase associated with germination) correlated well with spore cAMP content. Young wild-type spores, incapable of spontaneous germination, maintained a uniformly high cAMP level, and spore cAMP levels also remained high if germination was inhibited. When activated spores were deactivated by applying increased osmotic pressure, cAMP concentrations rose and ultimately levelled off at the high levels typical of dormant spores. The correlation between high cAMP and failure to germinate was also evident when autoactivation was inhibited by the cAMP analogue, 8-bromo-cAMP. Also, spores from a strain (HTY217) with unrestrained protein kinase A activity were incapable of spontaneous germination. Overall, our experiments provide evidence for continued cAMP signalling in spores up to 18 d after sporulation and for linkages between elevated cAMP, spore deactivation and inhibition of spontaneous germination. PMID- 10463155 TI - Ammonium phosphate in sori of Dictyostelium discoideum promotes spore dormancy through stimulation of the osmosensor ACG. AB - The sori of Dictyostelium discoideum (strains SG1, SG2, NC4 and V12) contained more than 100 mM ammonium phosphate. Glutamine synthetase (GS), which could remove ammonia from the sorus, was not present in 2-d-old dormant spores but enzyme activity returned to vegetative levels after spore germination. Based on mRNA blotting, the activity of this enzyme in germinating spores appeared to be transcriptionally controlled. At the same time that GS activity was increasing, ammonia was released from germinating spores. Exogenous ammonium ions at a concentration of 28 mM did not block germination nor modulate GS activity in nascent amoebae. It was concluded that the transcription and translation of GS is not environmentally regulated but is an integral part of the germination process, preparing nascent amoebae for vegetative growth. An exogenous concentration of 69 mM ammonium phosphate could maintain dormancy in spores of strains SG1 and SG2 for at least a week in the absence of any other inhibitory component from the sori. The inhibition was reversible at any time either by dilution or by washing the spores free of the ammonium ion. Spores of strain acg- were not inhibited by 100 mM ammonium phosphate. A model is presented in which GS in prespore cells serves as a sink for ammonia to allow the osmotically sensitive adenylyl cyclase aggregation protein (ACA) to activate protein kinase A (PKA) to induce fruiting body formation. After fruiting-body formation is complete, the decline in GS and ACA activities in developing spores is offset by their replacement with the osmotically and ammonia-stimulated adenylyl cyclase osmosensor for germination (ACG). Ammonia and discadenine may act as separate signals to synergistically activate PKA by stimulating ACG activity while inhibiting cAMP phosphodiestrase activity in fully dormant spores. PMID- 10463156 TI - Restoration of inositol prototrophy in the fission yeast Schizosaccharomyces pombe. AB - The biosynthesis of inositol requires only two enzymes, inositol-1-phosphate synthase (encoded by INO1) and an inositol monophosphatase, but the regulation of inositol biosynthesis is under multiple controls and is exquisitely regulated. In the budding yeast Saccharomyces cerevisiae, mutations in any of 26 different genes lead to inositol auxotrophy. The fission yeast Schizosaccharomyces pombe, however, is a natural inositol auxotroph. An investigation has been initiated to examine the possible reasons that might have led to inositol auxotrophy in Sch. pombe. Complementation with a genomic library of an inositol prototrophic yeast indicated that a Pichia pastoris INO1 gene alone could confer inositol prototrophy to Sch. pombe and that the gene was absent in Sch. pombe. To investigate possible reasons for the loss of INO1 gene in Sch. pombe, an attempt was made to disrupt inositol homeostasis in Sch. pombe by overproduction of intracellular inositol, but this did not lead to any discernible adverse effects. The sources of inositol in the natural environment of Sch. pombe were also examined. As the natural environment of Sch. pombe contains significant amounts of phytic acid (inositol hexaphosphate), an investigation was carried out and it was discovered that Sch. pombe can utilize phytic acid as a source of inositol under very specific conditions. PMID- 10463157 TI - The role of malic enzyme in the regulation of lipid accumulation in filamentous fungi. AB - The hypothesis is advanced that NADP(+)-malic enzyme (ME; EC 1.1.1.40) is an important activity in regulating the extent of lipid accumulation in filamentous fungi. In Mucor circinelloides, a fungus capable of accumulating only 25% (w/w, dry wt) lipid, even under the most propitious conditions, ME disappears 15-20 h after nitrogen exhaustion, coincident with the cessation of lipid accumulation. In contrast, ME in Mortierella alpina, a fungus capable of accumulating 50% (w/w, dry wt) lipid, remains active for over 60 h after N-exhaustion during which time lipid accumulation continues. No other enzyme activity studied, including the lipogenic enzymes acetyl-CoA carboxylase, fatty acid synthase, diacyglycerol acyltransferase, ATP: citrate lyase and the NADPH-generating enzymes glucose-6 phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and NADP+:isocitrate dehydrogenase, demonstrated any correlation with the accumulation of storage lipid in either fungus. Full activity of ME is restored in Mr. circinelloides within 4 h by adding NH4+ to the cultures, but this is prevented by adding cycloheximide as an inhibitor of protein synthesis. This suggests that the decrease in ME activity occurs due to down-regulation of the ME gene. PMID- 10463158 TI - Cryptocandin, a potent antimycotic from the endophytic fungus Cryptosporiopsis cf. quercina. AB - A unique lipopeptide antimycotic, termed cryptocandin, is described from Cryptosporiopsis cf. quercina, an endophytic fungus. Cryptocandin, with a molecular mass of 1079 Da, contains equimolar amounts of 3,4 dihydroxyhomotyrosine, 4-hydroxyproline, threonine, glutamine, 3-hydroxy-4 hydroxymethylproline, 4,5-dihydroxyornithine and palmitic acid. Cryptocandin is chemically related to well-known antimycotics, the echinocandins and pneumocandins, which are produced by such fungi as Zalerion arboricola, Pezicula spp. and Aspergillus spp. Cryptocandin has minimal inhibitory concentration values of 0.03-0.07 microgram ml-1 against isolates of Candida albicans, Trichophyton mentagrophytes and Trichophyton rubrum. Cryptocandin is also active against a number of plant-pathogenic fungi including Sclerotinia sclerotiorum and Botrytis cinerea. PMID- 10463159 TI - Spore surface glycoproteins of Colletotrichum lindemuthianum are recognized by a monoclonal antibody which inhibits adhesion to polystyrene. AB - Conidia (spores) of Colletotrichum lindemuthianum, a fungal plant pathogen causing bean anthracnose, adhere to the aerial parts of host plants to initiate the infection process. These spores possess a fibrillar 'spore coat' as well as a cell wall. In a previous study a mAb, UB20, was raised that recognized glycoproteins on the spore surface. In this study UB20 was used to localize and characterize these glycoproteins and to investigate their possible role in adhesion. Glycoproteins recognized by UB20 were concentrated on the outer surface of the spore coat and, to a lesser extent, at the plasma membrane/cell wall interface. Extraction of spores with hot water or 0.2% SDS resulted in removal of the spore coat. Western blotting with UB20 showed that a relatively small number of glycoproteins were extracted by these procedures, including a major component at 110 kDa. Biotinylation of carbohydrate moieties, together with cell fractionation, confirmed that these glycoproteins were exposed at the surface of the spores. In adhesion assays, > 90% of ungerminated conidia attached to polystyrene Petri dishes within 30 min. UB20 IgG at low concentrations inhibited attachment in an antigen-specific manner. This suggests that the glycoproteins recognized by this mAb may function in the initial rapid attachment of conidia to hydrophobic substrata. Polystyrene microspheres bound selectively to the 110 kDa glycoprotein in Western blots, providing further evidence that this component could mediate interactions with hydrophobic substrata. PMID- 10463160 TI - Isolation of the gene encoding an immunodominant membrane protein of the apple proliferation phytoplasma, and expression and characterization of the gene product. AB - An immunodominant membrane protein (IMP) of the apple proliferation (AP) phytoplasma was detected in preparations from AP-diseased periwinkle plants using monoclonal and polyclonal antibodies to the AP agent. Following isolation from Western blots and partial sequencing, degenerate oligonucleotides derived from the IMP sequence were used as probes to identify a DNA fragment containing the ORF encoding the IMP. Complete sequencing and subsequent analysis of the cloned DNA fragment revealed the presence of two ORFs, predicted to encode proteins with molecular masses of 25 kDa (P-318A) and 19 kDa (P-318B). Whilst database searches failed to assign a possible function to P-318A, analysis of P-318B predicted an amphiphilic membrane protein with a positively charged N-terminal portion, followed by a hydrophobic segment forming an alpha-helix, and a hydrophilic C terminal part located outside of the cell. The amphiphilic nature of P-318B was confirmed by its solubility in Triton X-114. The gene encoding P-318B was expressed in Escherichia coli and the resulting protein was used to immunize rabbits. The antiserum obtained reacted specifically with P-318B. The same protein was also detected by an antiserum raised against antigen preparations from AP-diseased plants. The P-318B antiserum did not react with antigen preparations from plants infected with the closely related pear decline phytoplasma. However, in Southern hybridization studies, the gene encoding the IMP hybridized to genomic fragments of the pear decline and European stone fruit yellows phytoplasmas. It also showed significant sequence similarity to a gene encoding an antigenic membrane protein of the sweet potato witches' broom phytoplasma, but not to a gene encoding a major immunogenic membrane protein of an aster yellows group phytoplasma. Since it appears that most phytoplasmas possess a major immunogenic membrane protein which may have a function in pathogenesis, this work may be a basis for further studies on fundamental aspects of host-pathogen interactions. It also describes a new approach to obtain suitable immunogens to produce specific antibodies for detection and characterization of the non-culturable phytoplasmas. PMID- 10463161 TI - Erwinia carotovora DsbA mutants: evidence for a periplasmic-stress signal transduction system affecting transcription of genes encoding secreted proteins. AB - The dsbA genes, which encode major periplasmic disulfide-bond-forming proteins, were isolated from Erwinia carotovora subsp. carotovora (Ecc) and Erwinia carotovora subsp. atroseptica (Eca), and the dsbC gene, encoding another periplasmic disulfide oxidoreductase was isolated from Ecc. All three genes were sequenced and mutants deficient in these genes were created by marker exchange mutagenesis. The Ecc mutants were severely affected in activity and secretion of pectate lyase, probably due to the absence of functional PelC, which is predicted to require disulfide bond formation to achieve its correct conformation prior to secretion across the outer membrane. Similarly, endopolygalacturonase, also predicted to possess disulfide bonds, displayed reduced activity. The major Ecc cellulase (CelV) does not contain cysteine residues and was still secreted in dsbA-deficient strains. This observation demonstrated unequivocally that the localization and activity of the individual components of the Out apparatus are independent of disulfide bond formation. Surprisingly, cellulase activity was shown to be increased approximately two- to threefold in the DsbA mutant. This phenomenon resulted from transcriptional up-regulation of celV gene expression. In contrast, transcription of both pelC and peh were down-regulated in dsbA deficient strains when compared to the wild-type. Protease (Prt) activity and secretion were unaffected in the Ecc dsbA mutant. Prt activity was considerably reduced in the double dsbA dsbC mutant. However Prt was secreted normally in this strain. The Eca dsbA mutant was found to be non-motile, suggesting that disulfide bond formation is essential for motility in this strain. All of the dsb mutants showed reduced tissue maceration in planta. These results suggest that a feedback regulation system operates in Ecc. In this system, defects in periplasmic disulfide bond formation act as a signal which is relayed to the transcription machinery regulating gene expression in diverse ways. PMID- 10463162 TI - Isolation of an extracellular protease gene of Erwinia carotovora subsp. carotovora strain SCC3193 by transposon mutagenesis and the role of protease in phytopathogenicity. AB - Using mini-Tn5CmR::gusA, a transposon that allows transcriptional fusions to a promoterless beta-glucuronidase gene, a mutant of Erwinia carotovora subsp. carotovora SCC3193 deficient in extracellular protease production and soft-rot pathogenicity in plants was isolated. The mutant, designated SCC6004, produced normal levels of pectate lyase, polygalacturonase and cellulase. The region of the transposon insertion was partially sequenced to permit the design of specific oligonucleotide primers to amplify a 2.7 kb Clal fragment from E. carotovora subsp. carotovora SCC3193. The DNA sequence of the cloned fragment contained two complete and one partial ORFs. One of the complete ORFs (ORF1) was designated prtW and encodes a secreted protease. The deduced amino acid sequence of PrtW showed a high overall identify of 60-66% to the previously described Erwinia chrysanthemi proteases, but no homology to other proteases isolated from different E. carotovora strains. Downstream from ORF1, a further complete ORF (ORF2) and a partial ORF (ORF3) were found, with deduced peptide sequences that have significant similarity to the Inh and PrtD proteins, respectively, from E. chrysanthemi, which are involved in protease secretion. Gene fusion to the gusA reporter was employed to charaterize the regulation of prtW. The prtW gene was found to be strongly induced in the presence of plant extracts. The mutant exhibited reduced virulence, suggesting that PrtW enhances the ability of strain SCC3193 to macerate plant tissue. PMID- 10463163 TI - A simplified subtractive hybridization protocol used to isolate DNA sequences specific to Xylella fastidiosa. AB - A simplified protocol of subtractive hybridization based on the technique of L. M. Kunkel, A. P. Monaco, W. Middlesworth, H. D. Ochs & S. A. Latt (1985, Proc Natl Acad Sci USA, 82, 4778-4782) was used to obtain DNA sequences specific to Xylella fastidiosa isolated from diseased citrus plants. As a driver, DNA extracted from bacteria showing different degrees of relatedness was used: Xy. fastidiosa 788 isolated from another host (plum), Xanthomonas campestris pv. campestris and Burkholderia gladioli strains. A DNA fragment, f14, showing no hybridization to the driver DNA, was used as a probe specific to Xy. fastidiosa from citrus and oleander. This fragment was sequenced and the predicted protein showed 40% similarity to the central region of flagellin of Escherichia coli serotypes H1 and H12. A pair of internal primers (f14-1 and f14-2) was designed for amplification of Xy. fastidiosa DNA. These primers detected Xy. fastidiosa strains isolated from citrus and oleander and yielded an amplification product of about 600 bp. They were also able to detect the bacteria in extracts from citrus plants with or without symptoms of disease. No amplification reaction was obtained using DNA extracted from other species and pathovars of Xanthomonas, Pseudomonas cichorii, Erwinia carotovora, Agrobacterium tumefaciens and phytopathogens of citrus (Xanthomonas axonopodis pv. citri) and coffee (Burkholderia andropogonis, P. cichorii, Pseudomonas syringae pv. garcae). The isolation of a DNA fragment specific to Xy. fastidiosa from citrus showed that the simplified protocol of subtractive hybridization used in this work is potentially applicable to other micro-organisms. PMID- 10463164 TI - Identification of novel bacterial lineages as active members of microbial populations in a freshwater sediment using a rapid RNA extraction procedure and RT-PCR. AB - A rapid method for the extraction of RNA from the indigenous bacterial communities in environmental samples was developed. The method was tested using anoxic sediment samples from a productive freshwater lake (Priest Pot, Cumbria, UK). The simple protocol yielded rRNA and mRNA of a purity suitable for amplification by reverse transcriptase PCR (RT-PCR). The integrity of the RT-PCR was demonstrated by amplifying 16S rRNA and mRNA for the mercury resistance regulatory gene merR. The diversity of 16S rRNA sequences recovered from RNA and DNA extracted from anoxic Priest Pot sediments was analysed. The 5' end of extracted 16S rRNA was amplified by RT-PCR and the 16S rRNA PCR products were cloned and sequenced to identify active constituents of the sediment bacterial community. Corresponding analyses were performed upon DNA templates from the same sediment samples. Partial 16S rRNA sequences were obtained from a total of 147 clones (71 rRNA-derived and 76 rDNA-derived). The clone libraries included sequences related to Pirellula staleyi, an aerobic planktonic member of the Planctomycetales, and the recently described candidate bacterial division OP5. Sequences from these groups were recovered in libraries generated from a DNA template but were also present in RNA-derived libraries. Previous studies of anoxic environments have identified sequences most closely related to Pirellula spp. This study, which utilized RT-PCR of 16S rRNA, has provided the first evidence that Pirellula-like bacteria are active in situ in an anoxic environment. Furthermore, members of the recently described candidate division, OP5, were also identified as active constituents of the bacterial community of anoxic Priest Pot sediments. This not only supports the widespread occurrence of OP5 members in diverse environments but suggests that they are active under anoxic conditions. PMID- 10463165 TI - Secretion of FK506/FK520 and rapamycin by Streptomyces inhibits the growth of competing Saccharomyces cerevisiae and Cryptococcus neoformans. AB - FK506 and rapamycin are immunosuppressants that inhibit signalling cascades required for T-cell activation, yet both are natural products of Streptomyces that live in the soil. FK506 and rapamycin also have potent antimicrobial activity against yeast and pathogenic fungi, suggesting a natural role in inhibiting growth of competing micro-organisms. The immunosuppressive and antimicrobial activities of FK506 and rapamycin are mediated by binding to the FKBP12 prolyl isomerase and the resulting FKBP12/FK506 and FKBP12/rapamycin complexes inhibit conserved protein targets, either the phosphatase calcineurin or the TOR (target of rapamycin) kinases, respectively. Streptomyces sp., 'Streptomyces hygroscopicus subsp. ascomyceticus' and Streptomyces hygroscopicus, which produce FK506, FK520 (also known as ascomycin, a C21 ethyl derivative of FK506) and rapamycin, respectively, produced toxins that inhibited the growth of competing cells of the yeast Saccharomyces cerevisiae and the pathogenic fungus Cryptococcus neoformans. Yeast and fungal mutants lacking FKBP12 or expressing dominant drug-resistant calcineurin or TOR mutants were resistant to FK506 and rapamycin, and to the toxins produced by Streptomyces. Streptomyces strains with mutations in the FK506 or rapamycin biosynthetic enzymes were impaired in toxin production. Finally, the toxins secreted by 'S. hygroscopicus subsp. ascomyceticus' and S. hygroscopicus promoted formation of FKBP12/calcineurin and FKBP12/TOR complexes in a two-hybrid assay and mutations that rendered calcineurin or TOR drug-resistant prevented interaction. These observations support the hypothesis that Streptomyces evolved to secrete FK506, FK520 and rapamycin as toxins to inhibit the growth of competing yeast and fungi. PMID- 10463166 TI - A comparison of the kinetics of plasmid transfer in the conjugation systems encoded by the F plasmid from Escherichia coli and plasmid pCF10 from Enterococcus faecalis. AB - Quantitative measurements of horizontal DNA transfer are critical if one wishes to address questions relating to ecology, evolution and the safe use of recombinant bacteria. Traditionally, the efficiency of a conjugation system has been described by its transfer frequency. However, transfer frequencies can be determined in many ways and may be sensitive to physical, chemical and biological conditions. In this study the authors have used the mechanistic similarity between bacterial conjugation and simple enzyme catalysis in order to calculate the maximal conjugation rate (Vmax) and the recipient concentration (K(m)) at which the conjugation rate is half its maximal value, for two different conjugation systems: the F plasmid from Escherichia coli and plasmid pCF10 from Enterococcus faecalis. The results are compared with the data obtained from the aggregation-mediated conjugation system encoded on pXO16 from Bacillus thuringiensis. The conjugation systems analysed are fundamentally different; however, they have some characteristics in common: they are able to sustain conjugative transfer in liquid medium and the transfer efficiencies are very high. Conjugation encoded by the F plasmid in E. coli involves the formation of small aggregates (2-20 cells), established by sex pili, and the plasmid's maximal conjugation rate was estimated to be approximately 0.15 transconjugants per donor per minute. Pheromone-induced conjugation in Ent. faecalis, which involves the formation of large aggregates, was found to proceed at a maximal conjugation rate of 0.29 transconjugants per donor per minute. Also, the K(m) value differed significantly between these conjugation systems; this may reflect the inherent differences in mating pair formation and transfer mechanisms. In these conjugation systems, the donors underwent a 'recovery period' between rounds of conjugative transfer and newly formed transconjugants required a period of about 40-80 min to mature into proficient donors. PMID- 10463167 TI - Involvement of the N- and C-terminal domains of Mycobacterium tuberculosis KatG in the protection of mutant Escherichia coli against DNA-damaging agents. AB - The Mycobacterium tuberculosis KatG enzyme, like most hydroperoxidase I (HPI) type catalases, consists of two related domains, each with strong similarity to the yeast cytochrome c peroxidase. The catalase-peroxidase activity is associated with the amino-terminal domain but currently no definite function has been assigned to the carboxy-terminal domain, although it may play a role in substrate binding. This paper reports another possible function of the KatG protein involving protection of the host cell against DNA-damaging agents. The M. tuberculosis katG gene, the 5' domain and the 3' domain were cloned separately, in-frame with the maltose-binding protein, into the vector pMAL-c2. These constructs were introduced into four DNA-repair mutants of Escherichia coli, DK1 (recA), AB1884 (uvrC), AB1885 (uvrB) and AB1886 (uvrA), which were then tested for their ability to survive treatment with UV light (254 nm), hydrogen peroxide (1.6 mg ml-1) and mitomycin C (6 micrograms ml-1). All three constructs conferred resistance to UV upon the recA E. coli cells, whereas resistance to mitomycin C was found in all repair mutants tested. Protection against hydrogen peroxide damage was less pronounced and predominantly found in the recA host. These results indicated that the M. tuberculosis katG gene can enhance DNA repair in E. coli, and that the 5' and 3' domains can function separately. UV sensitivity tests on Mycobacterium intracellulare and M. tuberculosis strains mutant in katG revealed that the katG gene product does not play an additive role in the survival of mycobacterial cells after exposure to short-wavelength UV irradiation, in repair-competent cells. PMID- 10463168 TI - Serotype 14 variants of the Spanish penicillin-resistant serotype 9V clone of Streptococcus pneumoniae arose by large recombinational replacements of the cpsA pbp1a region. AB - The high prevalence of penicillin resistance among Streptococcus pneumoniae isolates from Uruguay has been associated with the emergence of a penicillin resistant clone of serotype 14. Isolates of this clone were identical by multilocus sequence typing to members of the Spanish penicillin-resistant serotype 9V clone and possessed indistinguishable forms of the penicillin-binding protein 2b and 2x genes. Their pbp1a genes were also identical, except at the 3' end. The serotype 14 isolates were shown to be a variant of the Spanish serotype 9V clone which arose by a 22.2 kb recombinational replacement that introduced the capsular biosynthetic locus, and part of the neighbouring pbp1a gene, from a serotype 14 isolate. One end of the recombinational replacement was within the first gene of the capsular polysaccharide operon, cpsA, as the sequence of the upstream dexB gene, through most of cpsA, was identical in the penicillin resistant serotype 9V and 14 isolates, but the sequences differed in the rest of cpsA and in cpsB. The other recombinational junction was at the end of the divergently transcribed pbp1a gene, which is approximately 11.6 kb downstream of the capsular biosynthetic locus. Isolates of this serotype variant were also detected in Spain and Denmark. Penicillin-resistant serotype 14 isolates from Poland were also closely related to the penicillin-resistant serotype 9V clone, but have emerged independently, as one end of the recombinational replacement was upstream of dexB and the other was within pbp1a, but at a different position from that in the serotype 14 variants from Uruguay, Spain and Denmark. Serotype 14 variants of the Spanish serotype 9V clone have therefore arisen on more than one occasion by large recombinational replacements that extend from the start of the cps region into the pbp1a gene. PMID- 10463169 TI - Tracking adhesion factors in Staphylococcus caprae strains responsible for human bone infections following implantation of orthopaedic material. AB - Ten Staphylococcus caprae strains isolated from four patients and responsible for bone infections following implantation of orthopaedic material were compared to four S. caprae strains collected from milk samples of healthy goats. The following characteristics were investigated: Smal patterns, hybridization patterns with pBA2 (ribotypes), slime production, adhesion to matrix proteins (fibrinogen, fibronectin, collagen) and the staphylococcal adhesion genes (fnbA, clfA, cna, atlE, ica, fbe). None of the characteristics enabled us to distinguish the human strains from the goat strains. Slime was occasionally produced by S. caprae strains but all of them carried nucleotide sequences hybridizing at low stringency with the following genes: atlE encoding a S. epidermidis autolysin binding vitronectin and responsible for the primary adhesion to polystyrene, ica operon involved in the biosynthesis of a S. epidermidis extracellular polysaccharide, and the part of clfA encoding the serine-aspartate repeated region of a S. aureus cell-wall fibrinogen-binding protein. PMID- 10463170 TI - Helicobacter pylori VacA cytotoxin associated with the bacteria increases epithelial permeability independently of its vacuolating activity. AB - Polarized epithelial monolayers of Madin-Darby canine kidney (MDCK) cells were used to study the pathogenicity of Helicobater pylori, with an emphasis on the effect of VacA. The adherence of H. pylori to MDCK monolayers resulted in a decrease in trans-epithelial resistance (TER) across the cell monolayer. Isogenic vacA mutants did not lower the TER, demonstrating that the effect is strictly linked to the action of the toxin. A similar effect was observed with all VacA producing strains, including those producing m2 toxins that are inactive in the vacuolating assay. In contrast to that seen with purified toxin, TER decrease was not enhanced by acid pH, which may indicate that the toxin associated to the bacterial surface is possibly in a monomeric state and therefore does not require a pH-induced conformation to be active. These data raise the possibility that one role of VacA in ulcerogenesis may consist of increasing the paracellular permeability of the gastric epithelium. PMID- 10463171 TI - Membrane vesicles derived from Pseudomonas aeruginosa and Shigella flexneri can be integrated into the surfaces of other gram-negative bacteria. AB - Incubation of intact Salmonella typhi Ty21a, Salmonella enterica serovar Typhimurium (Salmonella typhimurium) aroA or Escherichia coli DH5 alpha with membrane vesicles (MVs) derived from either Shigella flexneri M90T or Pseudomonas aeruginosa dsp89 resulted in a significant incorporation of vesicle antigens into the outer membrane of the bacteria; each recipient strain possessed a surface mosaic of new Shigella and Pseudomonas antigens intermixed with the native antigens of the Salmonella or Escherichia strains. Electron microscopy of preparations during the integration of vesicle antigens revealed that the MVs rapidly fused with the outer membrane of the host strains. Western blot analysis of host bacteria confirmed the integration of foreign antigens. Quantitative analysis for binding and fusion of antigens using an ELISA showed that approximately 78.7 +/- 12.8 ng of the Pseudomonas and 67.5 +/- 13.8 ng of the Shigella LPSs (microgram host protein)-1 were integrated into the Sal. typhimurium strain. Similar integrations of the Shigella or Pseudomonas vesicles were found with the E. coli or Sal. typhi strains. There was no loss of viability in the recipient bacteria after incorporation of the MVs, although vesicle antigens became diluted during continued growth as daughter cells shared the vesicle antigens. The new antigens were highly stable after being incorporated into recipient strains, being able to withstand storage of several months at 4 degrees C as well as several cycles of freezing and thawing. Since the recipient bacteria are common vaccine strains, the procedure described here offers a simple efficient means of introducing exogenous surface antigens, in their native form, into the outer membranes of Gram-negative bacteria for possible vaccine use. PMID- 10463172 TI - A chimaeric plant virus vaccine protects mice against a bacterial infection. AB - The plant virus cowpea mosaic virus (CPMV) is an efficient carrier of foreign peptides for the generation of strong humoral immune responses. Peptides derived from both viruses and bacteria are strongly immunogenic when displayed on the surface of CPMV and elicit high titres of peptide-specific antibody. However, the protective effects of antibodies generated using bacterial epitopes in this system have yet to be demonstrated. In this study the ability of chimaeric virus particles (CVPs) to afford protection against bacterial infection was assessed. Immunization of outbred mice with CPMV expressing a peptide derived from outer membrane protein F of Pseudomonas aeruginosa (CPMV-PAE5) generated high titres of P. aeruginosa-specific IgG that opsonized the bacteria for phagocytosis by human neutrophils and afforded protection upon challenge with two different immunotypes of P. aeruginosa in a model of chronic pulmonary infection. When examined 8 d after challenge, CVP-immunized mice had fewer severe lung lesions and fewer bacteria in their lungs compared to mice immunized with wild-type virus. Different levels of protection were seen with CPMV-PAE5 when Freund's or alum adjuvants were used. These studies highlight the ability of CVPs to generate protective immunity against infectious disease agents. PMID- 10463173 TI - Antigenic variants in Bordetella pertussis strains isolated from vaccinated and unvaccinated children. AB - Bordetella pertussis shows polymorphism in two proteins, pertactin (Prn) and the pertussis toxin (PT) S1 subunit, which are important for immunity. A previous study has shown antigenic shifts in these proteins in the Dutch B. pertussis population, and it was suggested that these shifts were driven by vaccination. The recent Italian clinical trial provided the opportunity to compare the frequencies of Prn and PT S1 subunit variants in strains isolated from unvaccinated children, and from children vaccinated with two acellular and one whole-cell pertussis vaccine. Four Prn variants (Prn1, Prn2, Prn3 and Prn5) were found in the 129 strains analysed. Prn1, Prn2 and Prn3 have been described previously, whereas Prn5 is a novel variant. Prn1, Prn2, Prn3 and Prn5 were found in, respectively, 6, 41, 51 and 2% of the strains. The B. pertussis strains used to produce the vaccines administered in the clinical trial were found to produce Prn1, or a type which differed from Prn1 in one amino acid. The frequency of the Prn1 variant was found to be lowest in the strains isolated from vaccinated groups, suggesting that Prn1 strains are more affected by vaccine-induced immunity than Prn2 and Prn3 strains. Only one PT S1 type (S1A) was observed in the examined strains, which was distinct from the types produced by the vaccine strains (S1B and S1D). The S1A type also predominates in the Dutch B. pertussis population. The genetic relationship among B. pertussis strains analysed by IS1002-based DNA fingerprinting revealed that three fingerprint types predominate, representing more than 70% of the strains. Prn2 strains showed a greater variety of fingerprint types compared to Prn3, suggesting that Prn3 has emerged more recently. The results are discussed in the light of vaccine-driven evolution. PMID- 10463174 TI - Use of primate model system to identify Chlamydia trachomatis protein antigens recognized uniquely in the context of infection. AB - A primate model system was used to identify Chlamydia trachomatis antigens uniquely recognized in the context of infection. Serum antibody titres were measured in cynomolgus monkeys challenged urethrally with C. trachomatis serovar L2 elementary bodies (EBs). High-titre sera from these primates were used, in parallel with antisera against killed C. trachomatis EBs, to differentially screen an expression library of C. trachomatis serovar L2 DNA. Four clones were recognized only by antisera from infected monkeys. Sequence analysis revealed that three of these immunoreactive clones overlap a common ORF, designated ORF D242 (encoding p242), in the C. trachomatis genome database. The fourth clone contains two complete ORFs, each encoding 32 kDa proteins that share identity with Treponema pallidum TroA and TroB (ORFs D067 and D068 in the C. trachomatis database, respectively). Immunoblot analysis of Escherichia coli lysates expressing C. trachomatis TroA, TroB and p242 fusion proteins showed that p242 and TroA, but not TroB, were detected by the sera collected from infected primates. Antibodies directed at TroA and p242 were also detected in sera from several C. trachomatis-infected patients, demonstrating that these proteins are also recognized by humans following infection. Immunoblot analysis with antibody against TroA and p242 also demonstrated that both antigens are present in higher abundance in infected ChoK1 cells relative to purified C. trachomatis EBs. Immunofluorescence microscopy shows that TroA and p242 are both localized to intracellular developmental forms at the margins of growing inclusions. Collectively, these studies identify two C. trachomatis proteins that are under represented in EBs and are recognized uniquely in the context of infection. PMID- 10463175 TI - Mycoplasma synoviae surface protein MSPB as a recombinant antigen in an indirect ELISA. AB - Mycoplasma synoviae is a poultry pathogen causing respiratory disease and synovitis. A number of serological assays have been developed for diagnosis of M. synoviae infection; however, they lack sensitivity and/or are prone to false positive reactions. Using a combination of PCR and expression cloning, four overlapping regions (regions 1-4) of the surface antigen MSPB of M. synoviae WVU 1853 were expressed in a bacterial expression system. Immunostaining of the resultant polypeptides with chicken sera raised against different M. synoviae strains, or Mycoplasma gallisepticum S6, suggested that region 4 contained a highly antigenic and species-specific domain (amino acids 212-317) [corrected] of MSPB. A fusion protein of region 4 was expressed in the pMAL expression system and purified from cold-osmotic-shock fluids of Escherichia coli cells for use in an indirect ELISA. The potential of the purified antigen for detection of M. synoviae antibodies was assessed with sera obtained from chickens experimentally infected with different strains of M. synoviae or M. gallisepticum, or from uninoculated chickens. All the sera from M. synoviae-inoculated chickens provided higher absorbance values than those from M. gallisepticum-inoculated or uninoculated chickens. Chickens inoculated with M. synoviae 86079/7NS had detectable increases of serum anti-MSPB immunoglobulins at day 7 after inoculation. These studies have identified the most antigenic region of one of the major species-specific surface proteins of M. synoviae, and shown the potential of this protein as a serodiagnostic reagent. PMID- 10463176 TI - A gene family in Mycoplasma imitans closely related to the pMGA family of Mycoplasma gallisepticum. AB - The avian pathogen Mycoplasma gallisepticum possesses a large gene family encoding lipoproteins which function as haemagglutinins. Representative species of the pneumoniae phylogenetic group of mycoplasmas were examined for the presence of genes homologous to members of this multigene family. Antisera against the pMGA1.1 lipoprotein recognized a 35 kDa protein in Mycoplasma imitans, but did not recognize proteins of Mycoplasma genitalium, Mycoplasma pneumoniae, Mycoplasma pirum, Mycoplasma penetrans of Mycoplasma iowae in Western blots. A fragment of the pMGA 1.2 gene and oligonucleotide probes complementary to highly conserved coding and non-coding regions of pMGA genes bound to fragments of genomic DNA of M. imitans, but not to the genomes of M. genitalium, M. pneumoniae, M. pirum or M. penetrans, and only one probe bound to a fragment of the M. iowae genome. One homologue of the pMGA genes was amplified from the M. imitans genome by PCR and used as a probe to clone a 3.1 kbp DNA fragment from a library of HindIII-digested M. imitans genomic DNA. The contiguous DNA sequence of the PCR and HindIII clones was predicted to encode one complete and one partial ORF which shared some peptide sequence identity with the pMGA genes, including the signal peptidase II cleavage site and the proline-rich amino terminal region. Like the pMGA genes, the M. imitans genes were found to be members of a large gene family, with an association with GAA trinucleotide repeats, a feature which distinguishes these two families from the homologous vlhA gene family in Mycoplasma synoviae. The identification of these gene families in three phylogenetically distinct avian mycoplasma species, but not in human mycoplasmas, suggests their horizontal transfer between species infecting the same host. PMID- 10463177 TI - Characterization of apxIVA, a new RTX determinant of Actinobacillus pleuropneumoniae. AB - A fourth type of RTX determinant was identified in Actinobacillus pleuropneumoniae and was designated apxIVA. When expressed in Escherichia coli, recombinant ApxIVA showed a weak haemolytic activity and co-haemolytic synergy with the sphingomyelinase (beta-toxin) of Staphylococcus aureus. These activities required the presence of an additional gene, ORF1, that is located immediately upstream of apxIVA. The apxIVA gene product could not be detected in A. pleuropneumoniae cultures grown under various conditions in vitro; however, pigs experimentally infected with A. pleuropneumoniae serotypes 1, 5 and 7 started to produce antibodies that reacted with recombinant ApxIVA 14 d post-infection, indicating that apxIVA is expressed in vivo. In addition, sera from pigs naturally and experimentally infected with any of the serotypes all reacted with recombinant ApxIVA. The apxIVA gene from the serotype 1 A. pleuropneumoniae type strain Shope 4074T encodes a protein with a predicted molecular mass of 202 kDa which has typical features of RTX proteins including hydrophobic domains in the N terminal half and 24 glycine-rich nonapeptides in the C-terminal half that bind Ca2+. The glycine-rich nonapeptides are arranged in a modular structure and there is some variability in the number of modules in the ApxIVA proteins of different serotypes of A. pleuropneumoniae. The deduced amino acid sequences of the ApxIVA proteins have significant similarity with the Neisseria meningitidis iron regulated RTX proteins FrpA and FrpC, and to a much lesser extent with other RTX proteins. The apxIVA gene could be detected in all A. pleuropneumoniae serotypes and seems to be species-specific. Although the precise role of this new RTX determinant in pathogenesis of porcine pleuropneumonia needs to be determined, apxIVA is the first in vivo induced toxin gene that has been described in A. pleuropneumoniae. PMID- 10463178 TI - HmsT, a protein essential for expression of the haemin storage (Hms+) phenotype of Yersinia pestis. AB - The haemin storage (Hms) phenotype of Yersinia pestis has been shown to be involved in the blockage of fleas that is required for the transmission of plague from fleas to mammals. Previously, an operon encoding four genes, hmsHFRS, that are essential for the temperature-regulated Hms+ phenotype has been characterized. Here the isolation and characterization of a fifth gene, hmsT, that is essential for this phenotype is described. Conceptual translation of hmsT suggests it encodes a 44.8 kDa protein with a pl of 7.75. The gene for HmsT is located outside of the approximately 102 kb pgm locus of Y. pestis that contains the hmsHFRS operon. Hybridization studies indicate that Yersinia pseudotuberculosis but not Yersinia enterocolitica or Escherichia coli possesses a highly homologous gene. HmsT belongs to a family of PleD-related proteins with four highly conserved regions of homology. Although PleD is a regulator, the functions of the other members of this family have not been experimentally determined. The iron-responsive regulator, Fur, has previously been implicated in temperature regulation of the Hms phenotype. A good potential Fur-binding site (FBS) is located upstream of hmsT. Y. pestis M23 and two of five Y. pseudotuberculosis strains, which all exhibit a temperature-constitutive Hms phenotype, contain a 6 bp insertion in the putative FBS. E. coli MG1655 contains homologues of hmsHFRST (ycdSRQPT) but has an Hms- phenotype. Only ycdQ and ycdP complement mutations in their respective homologues, hmsR and hmsS, in Y. pestis. PMID- 10463179 TI - On-line monitoring of gene expression. AB - Gene expression in cultures of Escherichia coli has been determined in situ and on-line by the use of an electrochemical sensor. Intact bacteria were used to monitor the induction of the lacZ gene; the onset of stationary phase was also monitored, using a reporter gene fused to the RpoS-dependent promoter of the osmY gene. The technique described can in principle be used to determine the activity of any promoter, with a variety of reporter genes. This technology is non intrusive, allows real-time monitoring of gene expression, and will be useful in the study of growth regulation and development. PMID- 10463180 TI - Analysis of the CoIE1 stability determinant Rcd. AB - Multimer formation is an important cause of instability for many multicopy plasmids. Plasmid CoIE1 is maintained stably because multimers are converted to monomers by Xer-mediated site-specific recombination at the cer site. However, multimer resolution is not the whole story; inactivation of a promoter (Pcer) within cer causes plasmid instability even though recombination is unaffected. The promoter directs the synthesis of a short transcript (Rcd) which is proposed to delay the division of multimer-containing cells. Mapping of the 5' terminus of Rcd confirms that transcription initiates from Pcer. The 3' terminus shows considerable heterogeneity, consistent with a primary transcript of 95 nt being degraded via intermediates of 79 and 70 nt. Secondary structure predictions for Rcd are presented. Of four mutations which abolish Rcd-mediated growth inhibition, one reduces the activity of Pcer while the other three map to the rcd coding sequence and reduce the steady-state level of the transcript. RNA folding analysis suggests that these three mutant transcripts adopt a common secondary structure in which the major stem-loop differs from that of wild-type Rcd. A survey of 24 cer-like multimer resolution sites revealed six which contain Pcer like sequences. The putative transcripts from these sites have similar predicted secondary structures to Rcd and contain a highly conserved 15 base sequence. To test the hypothesis that Rcd acts as an anti-sense RNA, interacting with its target gene(s) through the 15 nt sequence, we used DNA hybridization and sequence analysis to find matches to this sequence in the Escherichia coli chromosome. Our failure to find plausible anti-sense targets has led to the suggestion that Rcd may interact directly with a protein target. PMID- 10463181 TI - A novel approach for the construction of a Campylobacter mutant library. AB - Given the lack of functional transposons for use in Campylobacter spp., an alternative method of insertional mutagenesis using natural transformation was developed. High efficiencies of transformation were only obtained with species specific DNA. This feature was a key element in the construction of mutant libraries of this bacterium. A chromosomal library of Campylobacter jejuni 81116 DNA was made in shuttle vector pUOA18. Next, a kanamycin-resistance (KmR) cassette was ligated into the inserts of the plasmids. C. jejuni 81116 was then transformed with the resulting products to allow homologous recombination between genomic fragments present in the shuttle vector and the chromosome. Transformants were pooled and chromosomal DNA from these transformants was used to retransform C. jejuni 81116. This resulted in transformants containing the KmR cassette in the chromosome but lacking the vector. In order to evaluate this approach for the construction of a mutant bank, the KmR insertional mutants were screened for loss of motility. Partial characterization of 11 non-motile mutants indicated that the inserted genes are involved in motility. Four mutants had the KmR cassette inserted in genes involved in flagella biosynthesis, namely flaA/B, neuB and flgK, and produced incomplete or no flagella. Four mutants had the KmR cassette inserted in genes possibly involved in flagella motor function: pflA, fliM and orf1 downstream of the fliN gene. Three mutants had the KmR cassette inserted in genes that are homologous to genes encoding hypothetical proteins of Helicobacter pylori. PMID- 10463182 TI - Genetic analysis of the GcvA binding site in the gcvA control region. AB - The GcvA protein both activates and represses the gcv operon and negatively regulates its own transcription. GcvA binds to three sites in the gcv control region and to one site in the gcvA control region; each of these binding sites contains the conserved 5 bp DNA sequence 5'-CTAAT-3'. This report describes the role this DNA sequence plays in autoregulation and expression of gcvA. Through single base-pair mutations, the importance of three of these five basepairs in the autoregulation of gcvA expression is shown. Two of the gcvA control region mutations described cause a gcvA::lacZ fusion to be overexpressed at 9-24 times the wild-type level. The increase in expression is due in part to a complete loss of autoregulation and in part to a GcvA-independent mechanism. One of the mutants was shown by Western blot analysis to increase the intracellular concentration of GcvA. This high level of gcvA expression subsequently causes the loss of purine mediated repression of a gcvT::lacZ fusion. However, overexpression of gcvR re established purine-mediated repression of the gcvT::lacZ fusion, supporting the model for gcv regulation that suggests the need for a relatively constant GcvA to GcvR ratio for appropriate regulation of gcv expression in response to glycine and purine availability. PMID- 10463183 TI - A multidomain xylanase from a Bacillus sp. with a region homologous to thermostabilizing domains of thermophilic enzymes. AB - The gene xynC encoding xylanase C from Bacillus sp. BP-23 was cloned and expressed in Escherichia coli. The nucleotide sequence of a 3538 bp DNA fragment containing xynC gene was determined, revealing an open reading frame of 3258 bp that encodes a protein of 120,567 Da. A comparison of the deduced amino acid sequence of xylanase C with known beta-glycanase sequences showed that the encoded enzyme is a modular protein containing three different domains. The central region of the enzyme is the catalytic domain, which shows high homology to family 10 xylanases. A domain homologous to family IX cellulose-binding domains is located in the C-terminal region of xylanase C, whilst the N-terminal region of the enzyme shows homology to thermostabilizing domains found in several thermophilic enzymes. Xylanase C showed an activity profile similar to that of enzymes from mesophilic micro-organisms. Maximum activity was found at 45 degrees C, and the enzyme was only stable at 55 degrees C lower temperatures. Xylotetraose, xylotriose, xylobiose and xylose were the main products from birchwood xylan hydrolysis, whilst the enzyme showed increasing activity on xylo oligosaccharides of increasing length, indicating that the cloned enzyme is an endoxylanase. A deletion derivative of xylanase C, lacking the region homologous to thermostabilizing domains, was constructed. The truncated enzyme showed a lower optimum temperature for activity than the full-length enzyme, 35 degrees C instead of 45 degrees C, and a reduced thermal stability that resulted in a complete inactivation of the enzyme after 2 h incubation at 55 degrees C. PMID- 10463184 TI - Transcription of genes near the sspE locus of the Bacillus subtilis genome. AB - The yfhP, yfhQ (mutY homologue), yfhS and yfhR (oxidoreductase homologue) genes, which are located upstream of the sspE locus, have been identified in the Bacillus subtilis genome. Transcriptional analysis showed that yfhP, yfhQ and yfhR are transcribed during the exponential growth phase, and sspE is monocistronically transcribed in the late sporulation phase and co-transcribed with yfhQ and/or yfhR during exponential growth. However, SspE was not translated during this period. Northern blot and primer extension analyses indicated that yfhS is transcribed by E sigma E during sporulation. No significant difference between wild-type and yfhS mutant strains was found in the rate of sporulation or germination, the heat tolerance of spores or the transcription of the sspE locus during sporulation. The transcription of the yfhP and yfhQ-yfhR-sspE loci increased 2.5- and 5.3-fold in a yfhP-deficient strain compared to the wild-type strain at t-2 (2 h before initiation of sporulation). In addition, transcription corresponding to the yfhR-sspE loci increased more than twofold with maximum values observed at t-1.5. These results suggest that YfhP may act as a negative regulator for the transcription of yfhQ, yfhR, sspE and yfhP. PMID- 10463185 TI - Transport mechanisms for cationic drugs and proteins in kidney, liver and intestine: implication for drug interactions and cell-specific drug delivery. PMID- 10463186 TI - The area under the effect-time curve as a target parameter for dosage adaptation in renal insufficiency. PMID- 10463187 TI - Pharmacokinetics and pharmacodynamics of insulin Lispro compared with regular insulin in haemodialysis patients with diabetes mellitus. PMID- 10463188 TI - Pharmacodynamic half-life and effect-time course in renal impairment. PMID- 10463189 TI - Cross-talk between activated tubular epithelia of human kidney and monocytes: a basis for target cell-specific pharmacotherapy? PMID- 10463190 TI - Role of the protein kinases A and C and of the calcium/calmodulin-dependent protein kinase II in the regulation of the renal basolateral PAH and dicarboxylate transporters. PMID- 10463191 TI - Identification by HPLC of uraemic retention solutes decreasing theophylline protein binding. PMID- 10463192 TI - Patterns of potassium (K) wasting in response to stepwise combinations of diuretics in nephrotic syndrome. PMID- 10463193 TI - Nephrotoxicity: focusing on radiocontrast nephropathy. PMID- 10463194 TI - Prostaglandin E1 (PGE1)--prophylaxis against radiocontrast-induced nephrotoxicity in patients with pre-existing renal dysfunction. PGE1 Study Group. PMID- 10463195 TI - Role of serotonin in the development of Chinese herbs nephropathy? PMID- 10463196 TI - Reconsideration of some hypotheses on the mechanism of thallium toxicity in rats with special respect to riboflavin and glutathione. PMID- 10463197 TI - Unexpected electrophysiological effects of D-19575, a new cytostatic drug. PMID- 10463198 TI - Glycosaminoglycan prevents hyperglycemia-induced renal TGF-beta 1 gene expression. PMID- 10463199 TI - Targeting TGF-beta overexpression: maximizing the antifibrotic actions of angiotensin II blockade in anti-Thy1 glomerulonephritis. PMID- 10463200 TI - Adenosine receptor antagonism in the prevention of acute cyclosporine A nephrotoxicity in normal, diabetic and hypertensive rats. PMID- 10463201 TI - Dopamine D3 receptors in the rat kidney: glomerular and tubular actions. PMID- 10463202 TI - Treatment with the angiotensin II antagonist valsartan in patients with chronic renal failure and hypertension. PMID- 10463203 TI - A randomized, double-blind, parallel study on the safety and antihypertensive efficacy of losartan compared to captopril in patients with mild to moderate hypertension and impaired renal function. PMID- 10463204 TI - Blood pressure reduction is necessary for the reduction of proteinuria in diabetic nephropathy--comparison of different antihypertensive agents. PMID- 10463205 TI - Protective effects of endothelin antagonists in chronic renal failure. AB - The present study suggests that ET-1 is involved in the pathogenesis of uraemic cardiac hypertrophy and in the progression of renal failure in rats with subtotal nephrectomy examined after an intermediate period of 12 weeks of renal failure. Furthermore, proteinuria is reduced by the selective ETA receptor antagonist more than by the unselective ETAB receptor antagonist, without reducing the blood pressure. ET receptor blockade might preserve renal function by reduction of protein excretion. In addition, ET receptor antagonists influence the aldosterone system. In our animal studies, the medication was well tolerated. Our study results provide a possible therapeutic approach using ET receptor antagonists for cardiac hypertrophy and renal protein excretion by blockade of endogenous ET-1. Further human studies are needed to show whether this protection of the heart and kidney might influence the survival and life expectancy of patients suffering from chronic renal failure, of patients on dialysis or after kidney transplantation. PMID- 10463206 TI - Decreased diurnal blood pressure variability and low dehydroepiandrosterone sulfate levels in patients with renal hypertension, and after kidney transplantation. PMID- 10463207 TI - Prostaglandin E1 reduces the risk of delayed graft function after cadaveric renal transplantation. PMID- 10463208 TI - Pharmacokinetics of mycophenolic acid (MPA) and free MPA in paediatric renal transplant recipients--a multicentre study. The German Study Group on Mycophenolate Mofetil (MMF) Therapy. PMID- 10463210 TI - Circulation and the kidney. Basic physiological and pathophysiological concepts. PMID- 10463209 TI - Therapeutic drug monitoring of total and free mycophenolic acid (MPA) and limited sampling strategy for determination of MPA-AUC in paediatric renal transplant recipients. The German Study Group on Mycophenolate Mofetil (MMF) Therapy. PMID- 10463211 TI - Interaction of the autonomic nervous and the renin-angiotensin system in heart and kidney. PMID- 10463212 TI - Diuretic therapy and diuretic resistance in cardiac failure. PMID- 10463213 TI - Treatment of cardiovascular changes in renal failure--ACE inhibition, endothelin receptor blockade or a combination of both strategies? AB - ACE-I and specific ETA receptor blockade comparably prevented the development of structural cardiovascular alterations such as LVH, myocardial interstitial expansion and wall thickening of intramyocardial and extracardiac arteries in experimental renal failure. However, the decrease in myocardial capillary supply and the concomitant increase in intercapillary distance could only be prevented by ETA receptor antagonism. These capillary changes which have been shown to occur in experimental renal failure as well as in uraemic patients play an important role in the pathogenesis of reduced cardiac ischaemia tolerance in renal failure. The data argue for a potential role of the local renin-angiotensin system as well as of the ET system in the pathogenesis of cardiovascular changes in renal failure and for ET receptor blockade as a new therapeutic option in the treatment of these alterations. In particular, myocardial capillary supply, which is particularly important for ischaemia tolerance in renal failure, seems to be modulated and regulated predominantly by ET-1. In contrast to what was found with respect to structural and functional changes of the kidney in various experimental models of renal damage, a combination therapy--at least in the doses used--does not seem to provide additional benefit in the prevention of cardiovascular changes compared with the respective monotherapies. PMID- 10463214 TI - Role of sympathetic nerves in the differential effects of T-type and L-type calcium channel blockers on renin secretion and renin gene expression. PMID- 10463215 TI - Anticoagulation with r-hirudin in a patient with acute renal failure and heparin induced thrombocytopenia. AB - After a Caesarean section subcutaneous bleeding with a secondary infection occurred accompanied by acute renal failure. Additionally the diagnosis of a heparin-induced thrombocytopenia type II could be made. Therefore, haemodialysis treatments were performed with r-hirudin as an alternative anticoagulation. Even in dialysis dependent acute renal failure a safe anticoagulation with r-hirudin is possible if aPTT and plasma r-hirudin are closely monitored. PMID- 10463216 TI - ATP release and degradation in the kidney: modulatory role of neuropeptide Y (NPY). PMID- 10463217 TI - Immunophenotyping and LDL receptor activity analysis in monocytes/macrophages during low- and high-flux haemodialysis. PMID- 10463218 TI - [Diagnostic value of regional cerebral blood flow in SPECT pattern in Alzheimer's disease]. AB - Regional cerebral blood flow in SPECT pattern was estimated in 20 cases of Alzheimer disease. In all patients diffuse hypoperfusion was found evidencing a great diagnostic value of SPECT. A special significance has the study of regional cerebral blood flow in the differential diagnosis of Alzheimer disease, frontal lobe dementia and pseudodementia in major depression. PMID- 10463219 TI - [Clinical symptomatology of primary brain atrophy]. AB - A group of 185 patients with brain atrophy found in CT is presented. The purpose of the study was to call attention to the the significance, frequency and clinical signs of brain atrophy. All patients were divided into two groups: 78 cases with secondary brain atrophy associated with other diseases of central nervous system and 107 cases of "primary" brain atrophy. The last one was an only finding without any other pathological lesions in the brain. Depending on the localisation, three groups of brain atrophy were isolated: generalized, subcortical and cortical. The clinical picture was analysed in the group of "primary" brain atrophy. In the studied group the most frequent cause of hospitalisation were epileptic seizures, the next one: headache, one-side hemiparesis or hemihypaesthesia, dizziness and incoordination. Neurological examination showed that one-side signs were observed very often in generalized brain atrophy. In the group of patients with cortical brain atrophy most patients were without any neurological signs. The analysis of the relationship between the clinical examination and "primary" brain atrophy found in CT, showed lack of characteristic, typical signs, which could be connected with brain atrophy. The problem of difficulty in the diagnosis of brain atrophy is stressed aesthesia. The definition and the significance of brain atrophy should be verified in diagnosis and the conclusions after finding brain atrophy have to be very careful. PMID- 10463220 TI - [Correlation of clinical symptomatology with the results of brain imaging in cases of leukoaraiosis]. AB - The aim of the study was establishing of correlations between the age of patients and their present and past diseases on one hand and the presence and intensity of leukoaraiosis (LA) on the other hand. The studied material comprised 91 patients hospitalized between Jan 1 1995 and June 15 1996 in whom features of LA were found on CT or MRI examination. Women and patients aged over 70 years prevailed in the group. Clinical analysis was based on neurological findings and routine diagnostic investigations. The intensity of LA lesions was assessed by means of a four grade scoring system known in literature. The analysis showed that higher grades of LA were more frequent on older patients: 14 had grade IV LA and 80 or more years of age. The greatest number of the patients had grade III LA and most of them were in the age range 70-80 years. Arterial hypertension was present in 64% of the patients, and more frequently it was associated with LA in younger patients. Stroke and myocardial infarction were given by 31% an 33% of the patients respectively in the history. Several had more than one stroke. Diabetes was found in 21 patients. Ischaemic brain episodes were the most cause of hospitalization. PMID- 10463221 TI - [Correlation between electronystagmographic findings and results of transcranial Doppler ultrasonography of vertebrobasilar arterial system in patients with vertigo]. AB - The purpose of the study was the determination of correlations between the results of electronystagmographic (ENG) investigations and the blood flow velocity in the vertebrobasilar arterial system measured by Doppler ultrasonography in patients with vertigo. The studied material comprised 68 patients (39 women and 29 men) aged 34-68 years (mean 52.4 years). ENG findings included unilateral hyporeflexia in 25 cases (37.7%), bilateral hyporeflexia in 15 (22.0%) and unilateral canal failure in 8 (11.9%). Doppler USG with neck rotation test showed normal flow velocity in 15 cases (22.0%) and pathological velocity in the remaining 53 cases (78.0%). These results showed that bilateral hyporeflexia is found usually in bilateral failure of vertebral arteries and unilateral hyporeflexia and unilateral canal failure were found with unilateral vertebral artery failure on the same side. PMID- 10463222 TI - [Molecular-genetic characteristics of mutations in dystrophin gene and clinical symptoms in Duchenne muscular dystrophy]. AB - The purpose of this paper is the estimation of the interrelationship of molecular findings with clinical studies on Duchenne muscular dystrophy (DMD), the estimation of molecular genetic findings efficiency focused on the diagnosis and the prognosis and carrier detection in relatives with recommendation of prenatal diagnosis possibilities. DNA isolated from peripheral blood lymphocytes of 100 patients was examined. DNA analyses was performed by multiplex PCR for promoter and 21 exons of DMD gene in regions where mutations are most frequent. Deletions were detected in 55% of the cases. In cases with no deletions detected, PCR-SSCP and PCR-HD analysis were performed in order to detect point mutations. For selected introns and exon 48 the occurrence of the previously described polymorphism was confirmed. Mutation causing formation of shortened protein was detected in exon 6 of two patients. Point mutation analysis is important complement of molecular diagnostics of Duchenne muscular dystrophy in patients with no deletions. For each family at risk of DMD the analysis of mutant allele was performed and carrier status evaluated. PMID- 10463223 TI - [Epileptic seizures in relation to the age of SSPE onset]. AB - The aim of this study is a clinical and electroencelographic analysis of those SSPE patients who suffered epileptic seizures in course of the disease. The material is based on an analysis of a computed database including 1180 case histories from multiple hospitalizations of 248 SSPE patients (141 males, 107 females) in years 1978-1995. The analysis was made using computer system EPI-INFO 6. The average age of SSPE onset was 12.3 +/- 4.5 (range 4-27.5). 74.2% of patients developed the disease before the age of 15. Epileptic seizures occurred in 43.5% (N = 108) of all SSPE patients in different stages of the disease. In 47 patients (M-21; F-26) they were observed among first SSPE symptoms, in 41 of those the seizures had the form of GTCS. Epileptic seizures were more frequent in patients with SSPE onset before the age of 15 (50.5%) than in patients with later onset (23.4%). More frequent occurrence of epileptic seizures and epileptiform changes in EEG in children under 15 can be associated with specificity of developmental age: immaturity of nervous and endocrine system and decreased convulsant threshold. PMID- 10463225 TI - [Tc-99m-MIBI and Tc-99m-HMPAO accumulation in primary and metastatic brain tumors assessed by brain SPECT]. AB - Imaging of intracranial lesions is frequently difficult in CT/MRI diagnosis. The introduction of the encephalic tracer MIBI seemed to be a promising alternative. We tried to compare the accumulation of Tc-99m-MIBI and HMPAO CT/MRI interpretationeally difficult intracranial lesions. We did SPECT scanning with single-head and triple-head gammacamera after i.v. administration of Tc-99m HMPAO/MIBI. We performed HMPAO scanning in 34 patients, MIBI scanning in 41 pts. CT and MRI scanning was performed simultaneously. Histological diagnosis was done intra- and/or postoperatively. Increased accumulation of Tc-99m-MIBI was seen in 7/7 meningiomas (100%), 7/9 gliomas (77%), 10/15 brain metastases (66%), 2/4 unverified. In 5 cases with finally non-neoplastic diagnosis (stroke, a-v malformation) no MIBI accumulation was seen. Increased accumulation of HMPAO was seen in 3/11 gliomas, 2/7 meningiomas, 2/8 metastases and 2/4 unverified. In 4 cases non-neoplastic diagnosis was established. Our results seem to be less promising, than reported so far, particularly for heterogeneous accumulation of MIBI in metastatic tumours. The coupled Tc-99m-MIBI/HMPAO scanning may be useful in primary tumour diagnosis, particularly in meningiomas. PMID- 10463224 TI - [Brain perfusion changes after head trauma assessed by cerebral SPECT with aminophylline test]. AB - Tomographic cerebral blood flow SPECT scanning using Tc-99m-HMPAO was performed before and after i.v. administration of aminophylline in 36 patients between 6 months to 11 years after closed head trauma. Asymmetries of brain perfusion were found in 34 patients, normal rCBF distribution in 2 patients. Bi-focused asymmetries on the presumed line: trauma-contra coup were seen in 64% of the patients. In 70.4% of the patients with brain perfusion asymmetries the CT scan was normal. The aminophylline test improved the perfusion in 42% of the patients, indicating preserved perfusion reserve. Persistence of brain perfusion alterations after head trauma exists. Aminophylline test may be useful in the assessment of brain perfusion reserve in post-traumatic focus. Brain perfusion SPECT scans may be useful for medico-legal purposes. PMID- 10463226 TI - [Macro EMG in neuromuscular diseases]. AB - The aim of this study is the introducing of macro-emg method as electrophysiological test used in diagnosis of neuromuscular diseases. The macro motor unit potentials (macro MUPs) obtained by recording macroelectrode (modified single-fibre electrode) represents temporal and spatial summation of individual single fiber action potentials belonging to whole motor unit territory--so the uptake area is larger for macroelectrode than for the concentric electrode, commonly used in emg routine work, when central main complex is generated only from less than 15 muscle fibers [10, 12, 13]. Additional information obtained by macro-emg method is spatial organisation of muscle fibers within the motor unit, so-called fiber density (F.D) In our study macro-emg examinations were performed in 20 healthy subjects, aged 21-55, without signs and symptoms of neuromuscular disorders. Macro MUPs were recorded using special programme for macro-emg and performed on electromyograph Counterpoint. 37 muscles (20 BB and 17 RF) were examined, and median values of amplitude, area of macro MUPs and F.D. in healthy subjects of different age were analyzed. Mean values of median for amplitude and area of macro MUPs in BB and RF muscles show respectively--148 microV, 382 microV x ms, and 319 microV, 763 microV x ms. Parameters of macro MUPs obtained in healthy subjects were compared to results obtained in 10 patients with myopathy and lower motor neuron lesion. Our results have confirmed the value of macro-emg method for investigating of the pathophysiological changes in motor units in neurogenic disorders, in myopathy the study should be continued. PMID- 10463228 TI - [Vertebral column primary neoplasms]. AB - Clinical analysis was carried out in 53 cases treated surgically because of primary vertebral column neoplasms in Department of Neurosurgery in Poznan between 1976-1995. Plasmocytoma solitaire (9 cases), angioma (5 cases), osteoblastoma (4 cases) and aneurysmal bone cysts (4 cases) were observed most frequently. Neoplasms localization was as follows: cranio-vertebral junction (4 cases), cervical segment (14 cases), thoracic (23 cases) and lumbo-sacral segments (12 cases). Severe neoplastic vertebral column destruction was disclosed in 34 (64%) patients, but complete transversal spinal syndrome was noted only in 7 (13%) cases. Surgical procedures: posterior and latero-posterior approaches were applied in most cases while anterior and lateral approaches--were rarely used. In 3 cases two stage operations were performed. Intervertebral fusion or posterior autogenous graft was necessary in 39 cases. PMID- 10463227 TI - [Hippel-Lindau disease]. AB - Hippel-Lindau disease is one of inherited tumour susceptibility syndromes. The most common lesions are located in central nervous system, retina and visceral organs. In Poland the disease was rarely diagnosed although the prevalence is much higher than it was supposed and is estimated as 1: 30-50,000. It is inherited in an autosomal dominant manner with age related penetrance reaching almost 98% penetrance at the age of 60 and variable expression. The VHL gene is located near the tip of the short arm of chromosome 3 (3p25-26). Classical lesions in VHL patients are: haemangioblastomas of CNS, retina, cysts and clear cell carcinoma of kidney, cysts and tumours of pancreas, phaeochromocytoma and paraganglioma, papillary cystadenoma of epididymis and endolymphatic sac tumours. Multifocal, often bilateral lesions in form of benign cysts, vascular tumours or carcinomas occur. Management of the lesions often differs from that in sporadic cases of the tumours. Non-symptomatic lesions of CNS need no treatment, neither do non-symptomatic tumours of epididymis and some of phaeochromocytomas. Kidney carcinoma is treated when it reaches a certain size preferably by nephron-sparing surgery. Special care should be provided to pregnant VHL patients. Available DNA testing enables to identify VHL carriers. Although the mean age of death in VHL patients is 41 at the moment a proper prophylactic, diagnostic and treatment management can probably prolong survival of the patients and limit complications of the disease. The coordination between genetic consultants and clinicians is crucial in the management of the patients. The authors coordinate work of Polish VHL Registry and Polish VHL Association. PMID- 10463229 TI - [The role of surgery in comprehensive treatment of spinal neoplasms]. AB - Basing on clinical experience of 212 surgically treated spinal tumours the authors present the principal problems in surgical treatment of neoplasmatic disease of the spine. The essential aim of surgical treatment is the tendency to the radical removal of the tumour and, in the same time, to achieve a good stabilization of the spine. The best method of stabilization of the spine is internal stabilization, transpedicular and interbody. The removed parts of the vertebral column should be replaced with bone grafts. The surgical treatment should be combined with oncological and rehabilitation. PMID- 10463230 TI - [Remote results of conservative treatment of traumatic optic neuropathy]. AB - The problem of pathogenesis and effective treatment of traumatic optic neuropathy (TON) is still an open question. According to current opinions the immediate results of conservative treatment with megadose corticosteroids are similar to those obtained with surgical decompression. No data on late results of both modes of treatment are at present available. This study was undertaken to assess the late results of conservative treatment of traumatic optic neuropathy. The clinical material comprised 15 patients (3 women and 12 men, age 14-64 years), who developed clinical symptoms of TON as a consequence of closed head trauma. All were treated conservatively with megadose steroid therapy. 10 patients presented to follow-up examination performed 3-11 years after the injury. A full ophthalmologic survey and colour-coded Doppler (CCD) examination of the orbital vessels were performed in all subjects. In 6 patients there was full blindness of the affected eye since the injury. Their visual acuity did not improve despite vigorous treatment and their eyes were still blind at the late follow-up examination. No flow found with CCD in their central retinal arteries at follow up might suggest structural disruption of the nerve. In 5 of these patients ocular atrophy evolved between the injury and follow-up examination. 4 patients, who on admission displayed merely light sensation, responded to 2-3 weeks of steroid therapy with improvement of visual acuity to 3/50, 5/50, 5/10, 5/7. After 4-6 years however, visual acuity of all these patients deteriorated again to mere light perception in three and 1/50 in another one. Optic atrophy was diagnosed in all the affected eyes. Our results indicate that conservative treatment of TON, even if giving satisfactory relief of the symptoms, may not be reliable in permanent restrainment of the sequela of traumatic optic neuropathy. PMID- 10463231 TI - [Clinical patterns of Arnold-Chiari malformations]. AB - The purpose of the study was clinical analysis of the patients with Arnold-Chiari malformation and the definition of the groups according to the clinical presentation. The authors present the series of 210 patients treated between 1980 and 1997 in the Department of Neurosurgery. These patients were classified into 4 groups: syringomyelia, cerebellar syndrome, cerebello-spinal syndrome and hydrocephalus. Syringomyelia was the commonest clinical presentation in the series. Less often cerebellar and cerebello-spinal syndromes were observed. Non communicating hydrocephalus was rare. The symptomatology of the Arnold-Chiari malformation appears extremely variable, but most often is related to the associated cavitation of the spinal cord. The signs and symptoms in patients presenting cerebellar syndrome may lead to misdiagnosis of insufficiency of the vertebrobasilar circulation. Diffuse involvement of the motor and sensory system with cerebello-spinal syndrome may cause the patient to be diagnosed incorrectly as having sclerosis multiplex. The MR is the method of choice in the diagnosis of Arnold-Chiari malformation. PMID- 10463232 TI - [Piracetam in severe cranio-cerebral injuries]. AB - A group of 100 patients treated immediately following a cranio-cerebral injury was analyzed. The patients, administered piracetam either in an intravenous infusion (GCS 3-8) or orally (GCS above 9), were divided into groups depending on the dose and clinical status. Piracetam participates in the activity of the majority of neurotransmitters, increases glucose and oxygen consumption in the ischaemic nervous tissue and increases blood flow through cerebral terminal vessels. In cranio-cerebral injuries, piracetam is employed to achieve cytoprotection and improve cerebral blood flow. In patients with neurological deterioration following the administration of 6-10 mg/day, no good results were obtained. A dose of 24-30 mg/day had a significant positive effect on therapeutic results providing certain conditions were met, such as ensuring proper partial oxygen pressure (oxygen therapy) and proper blood glucose levels. The use of piracetam is justified immediately after an injury; after the discharge oral piracetam therapy is recommended. PMID- 10463233 TI - [The examination of the blood flow velocity in arterial vessel during and after encephalo-dura-arterio-synangiosis. Preliminary results]. AB - The Japanese authors first described encephalo-duro-arterio-synangiosis, a new treatment for moya-moya disease, 17 years ago. The other authors used this method (EDAS, indirect anastomosis) in surgical treatment of patients suffered cerebral transient ischaemic attack and cerebral ischaemia. In our department this method was applied in 5 patients with cerebral ischaemia. All patients had internal carotid artery occlusion. After angiography, to localize carotid artery occlusion, acetazolamid (Diamox) test was performed. The patients with negative Diamox test were treated surgically (EDAS). The authors used the intraoperative microdoppler device to monitor blood flow velocity of the prepared vessel (temporal superficial artery) at every stage of surgery. The device was also useful in controlling the patency of encephalo-duro-arterio-synangiosis percutaneously after the surgery. The proximal and the distal part of the non bypass anastomosis was examined in the follow up during 5-8 months before angiography. PMID- 10463234 TI - [Mutation of presenilin ++ genes and their role in pathogenesis of Alzheimer's disease]. AB - A new family of presenilin genes involved in the pathogenesis of early-onset autosomal dominant Alzheimer's disease (AD) has been identified recently. Mutations in presenilin-1 and presenilin-2 genes have a full penetration and lead to AD. The presenilins are transmembrane proteins localized mainly within endoplasmatic reticulum and Golgi systems. Their biological function remains unknown. It has been suggested that the presenilins may be important for intracellular processes of protein trafficking and processing. Presenilin mutations play a direct role in the beta-amyloid precursor protein metabolism and cause increased production of amyloidogenic A beta 42(43) peptide. PMID- 10463235 TI - [Inflammatory changes and immunological disturbances in stroke]. AB - During recent years it has been demonstrated that activated leukocytes may play a role in disturbances of flow in microcirculation, such disturbances leading to the enlargement of the necrosis zone and area of ischaemic penumbra. Astrocytes, microglial and endothelium cells are also active in the evolution of ischaemic changes in brain. The above cells are producing cytokines, which upregulate the expression of adhesion molecules being responsible for accumulation of inflammatory cells in injured brain. Very significant is also the secretion of neurotoxic substances such as free radicals, nitric oxide or proteolytic enzymes by leukocytes and glial cells. Pre-stroke infection may increase activation of leukocytes and enhance inflammatory process in ischaemic focus. This paper discusses also the results of a treatment of the experimental brain stroke using therapy which reduces inflammatory and immunological response. PMID- 10463236 TI - [Spontaneous dissection of carotid and vertebral arteries. II. Vertebral arteries and their branches]. AB - Dissection of the extra- and ntracranial portions of vertebral arteries, as well as basilar artery and their branches are discussed based on the up-to-date literature. Formerly, arterial dissections were found exclusively at necropsy, yet they have currently arisen to the field of interest of not only neurologists but also radiologists, neurosurgeons, and vascular surgeons. Ischemic strokes frequently result from arterial dissection, in particular subintimal dissection (most often in the extracranial portion of vertebral artery), by formation of emboli or reduced perfusion. Conversely, subadventitial and transmedial dissections produce aneurysms (most often in the intracranial portion of vertebral arteries and basilar artery) which may result in subarachnoid or intracerebral hemorrhages. The other possible mechanism of focal lesions is the compression of adjacent structures by dissected artery, most frequently by dolichoectasia of basilar artery which originates from its multiple intramural lesions. The dissections may present with cranial nerve palsies, symptoms of ischemia of cervical spine, cerebellum, and cerebral trunk, as well as occipital lobe syndromes. Arterial dissection may occur in adolescents and young adults. The clinical symptom that should be stressed is headache which precedes the occurrence of stroke for several days. The right diagnosis gives a chance for proper treatment, including operative procedures, and improved prognosis. PMID- 10463237 TI - [Lumbar interbody fusion. Biomechanical significance for the spine]. AB - The authors discuss the significance of interbody fusion for early and long term stability of the lumbar spine. They stress that the aim of the modern spinal instrumentation is to promote bone healing and not to replace fusion. Without fusion every stabilization device will fail in fatigue. The biomechanics of different types of spinal fusion and biomechanical conditions at fusion site are discussed. A history of lumbar interbody fusion including threaded implants (cages) is presented. Interbody cages combine positive properties of tricortical bone graft: the strength of cortical to the bone with improved incorporation properties of cancellous bone. In contrast bone graft their biomechanical performance is far better: they increase strength and stability of osteosynthesis, do not go collapse and resorption, prevent from decrease in disc space height and kyphotic angulation of the fused motion segment, require less bone to achieve fusion. They can be used solely without support of any stabilization system. Preliminary experience in fusion with interbody threaded implants suggest significant efficacy of this method. PMID- 10463238 TI - [Familial dystonia: a case of idiopathic torsion dystonia]. AB - A 43 year old man suffered from the involuntary movements since 10-th year of age. Those movements, initially mild, have progressed in the course of the disease. In the neurological examination the dystonic involuntary movements of the neck, trunk and limbs, more expressed on the right side, were observed. The laboratory findings were normal, except the level of dopamine beta-hydroxylase, which was increased. Similar results were observed also in the closest relatives of our patient. After the treatment with high doses of L-Dopa we achieved a considerable decrease of the symptoms. PMID- 10463239 TI - [A case of myotonic dystrophy proved by DNA analysis]. AB - The authors describe a case of a 53 years old man with myotonic dystrophy. Based on characteristic clinical symptoms and EMG results the diagnosis was established and proved on DNA examination. Myotonic dystrophy gene analysis showed on 3' untranslated region one hundred and fifty CTG triplet repeats. The accessory examinations revealed: cataract arrhythmias, gallbladder stones, impotence, cerebral atrophy on neuroimaging (CT and MRI). Apathy, somnolence, concentration troubles were present. Pedigree of the presented patient and possibility of spontaneous mutation are discussed. PMID- 10463240 TI - [A case of laryngeal adductor dystonia treated with transcutaneous injections of botulinum toxin]. AB - We present a case of 47-year old patient with a rare form of focal dystonia restricted to laryngeal adductors with blepharospasm. Apart from typical symptoms of blepharospasm, the patient had severe problems with articulation in the form of harsh voice, frequently interrupted speech and the sound coming out with a great effort. We applied a transcutaneous botulin toxin therapy to this patient. The toxin was given into thyroarytenoid muscle in transcutaneous injections under control of EMG. Successful clinical effect was achieved with the dose of 80 U of the botulin toxin (Dysport) and lasted 6 months. The treatment was repeated, and the patient has not presented the symptoms of the disease since then for 10 months. We confirm that the botuline toxin transcutaneous injections represent a save and effective treatment for laryngeal adductor dystonia. PMID- 10463241 TI - [Lymphocytic meningitis with the involvement of the skull in the course of spinal cord neoplasm simulating neuroborreliosis. Case report]. AB - We describe a case of 48 year old male patient treated in the Department of Parasitic Diseases and Neuroinfections AMB with suspected neuroborreliosis. CLINICAL SYMPTOMS: lymphocytic meningitis with cranial neuropathies n. VII palsy and radiculitis after numerous tick bites in endemic area--indicated neuroborreliosis. Because there was no effect of antibiotic therapy and lack antibodies against B-burgdorferi in serum and CSF we excluded neuroborreliosis. Developing neuropathies III-XII, increasing cytosis and protein concentration, radiculalgia and difficulties in walking, cachexia made us think of tbc etiology. Patient failed to improve after anti-tbc treatment. CT and MR showed presence of neoplasmatic masses in spinal canal. In cytologic examination "neoplasma malignum male differentiatum probabiliter metastaticum" was found. Primary focus of neoplasmatic process was not found. PMID- 10463242 TI - [Micturition syncope: a peculiar cause of diagnostic doubt]. AB - A 22-year old healthy man lost consciousness during urination in standing after night's sleep. He fell down to the floor without convulsions. A physician present by chance found on bradycardia. A 24-hours ECG monitoring disclosed an almost permanent sinus bradycardia (50-55 per minute). CT showed an arachnoid cyst at the base of the left temporal lobe without mass effect. In EEG only single slow waves above left temporal region were revealed. It seems, the cyst was found by chance and had no any pathogenetic relevance to the syncope. However, the presence of a structural change of the brain had not been described in micturition syncope till now and this makes careful differentiation from epileptic seizure essential. During the following 2 years the patient was well and the episode did not recur again. The control CT and EEG remained unchanged. Data from literature about syncope in association with micturition also in older persons, in majority women with chronic illnesses, are cited. PMID- 10463243 TI - [Intradural lumbar disk herniation (ILDH). Case report and literature review]. AB - In this report, the author presents a rare case of a 59 year-old man with ILDH. It was the only case of ILDH among 960 patients surgically treated, during the period 1989-1996. Clinically the patient demonstrated an acute cauda equina syndrome. The diagnosis was established by radiculograms, which showed a total block at the L3-L4 level. There was a 3 days time interval between the diagnosed syndrome itself and the operation. At surgery the L3-L4 level was intact, whereas dense adhesions were found between the L4-L5 disc and the dura. Root retraction to expose the nucleus pulposus mass was impossible. A laminectomy of the L4 was undertaken. An incision was made in the dura and arachnoid, revealing an extruded discal mass, lying between the roots of the cauda equina. It was carefully removed. The state of the patient at follow-up 1 year after surgery was unsatisfactory. The patient has moderate flaccid paraparesis, bladder dysfunction improved. The prognosis appeared to be linked to the preoperative duration of symptoms. PMID- 10463244 TI - [Blood flow velocity in the anterior part of the circulus arteriosus in ischemic stroke: correlation with Doppler ultrasonography of the internal carotid arteries]. AB - The correlation was studied between blood flow velocity in the anterior part of the circulus arteriosus and the condition of the internal carotid arteriess in their extracranial course studied by Doppler USG. The arteries in the anterior part of the circulus were studied in 100 patients with ischemic stroke, and in 50 controls without cerebrovascular disease. Only strokes in the area of the middle cerebral artery (MSC) were considered. The patients were divided into 4 subgroups depending on the condition of the extracranial portion of the carotid internal arteries (ICA): 1. with normal pattern of arteries in USG-duplex, 2. with narrowing without haemodynamic importance, 3. with high-grade stenosis and haemodynamic disturbances, 4. with occlusion of the ICA. Transcranial Doppler USG was carried out through a "temporal window" assessing blood flow velocity in the MCA and in the anterior cerebral arteries (ACA) on both sides. RESULTS: In subgroups 1 and 2 the mean flow velocity in MCA was reduced on the side of the stroke. In groups 3 and 4, apart from reduced flow velocity in the MCA on the side of the stroke, increased velocity on both sides was found in the ACAs. CONCLUSION: The degree of ICA narrowing in its extracranial part was influencing the condition of intracranial circulation. PMID- 10463245 TI - [Presence of immune complexes in ischemic stroke]. AB - There is increasing evidence that inflammatory responses play an important role in the pathogenesis of atherosclerosis and cerebral infarct. The aim of this study was to determine the amount of immunocomplexes (i.c.) in patients with stroke in the early period of the disease. Studies were performed on 35 subjects. The concentration of immunocomplexes was determined with the precipitation method (3.5% polyethylenoglikol was used). Increased concentration of i.c. was found in patients with cerebral infarct (after 12 hours and 7 days). It suggests, that i.c. could be one of the markers for systemic inflammation and be important in the patogenesis of atherosclerosis and stroke. PMID- 10463246 TI - [Urine free light chains kappa in multiple sclerosis]. AB - The evaluation of free light chains kappa in urine was performed in 77 cases of multiple sclerosis, including 52 patients before and after treatment with 2 CDA and in 25 patients before and after therapy with high doses of prednisone. The high variations in the level of free kappa chains indicate a limited diagnostic value, and only for cases with very high level. We have found effect of 2 CDA therapy in chronic progressive MS group on free kappa light chain value. A significant effect of prednisone treatment was observed in early onset cases of multiple sclerosis and in cases with clinical improvement after therapy. In conclusion, the study suggests that urinary free light chains level may be considered as one of markers for monitoring of the effect of therapy on the activity of the immunological processes in multiple sclerosis. PMID- 10463247 TI - [Clinical-electrophysiological characteristics of Alzheimer's dementia and cerebrovascular dementia based on determination of cerebral refraction time]. AB - The aim of the study was an electrophysiologic assessment of cerebral refractory period in relation to chosen clinical parameters, EEG and CT in patients with dementia of various origin. The study included 30 patients (11 female, 19 male) aged 50-75: 15 with dementia of Alzheimer type (DAT) and 15 with vascular dementia (VD), and 15 age-matched healthy controls. Cerebral refractory period (CRP) was assessed by means of somatosensory evoked potentials with the use of paired stimulation with varying interstimulus interval, applied to the median nerve. Interstimulus interval, with which one somatosensory response was obtained to paired stimuli, was determined as absolute refractory period. CRP was significantly prolonged in demented patients, when compared to the control group, with no significant difference between groups with DAT and VD. CRP correlated negatively with age and positively with Mini Mental State Examination results, assessing the degree of mental deterioration. Prolonged CRP was noted in patients with vast lesions shown in CT. No relationship between CRP and EEG changes was found. CRP is a sensitive parameter, which may contribute to electrophysiological assessment of cerebral function. PMID- 10463248 TI - [Cerebrospinal fluid serotonin concentration in pyogenic and tick-borne encephalomeningitis]. AB - Cerebrospinal fluid (CSF) serotonin was determined in 38 patients with pyogenic meningitis and tick-borne encephalitis (TBE) before and after treatment. Increase of CSF serotonin concentration was observed in acute phase of pyogenic meningits and normalized after treatment. PMID- 10463249 TI - [Spasmodic torticollis: experience of 5 years with botulinum toxin treatment]. AB - 64 cases with spasmodic torticollis were observed during 5 years and treated with botulinum toxin (BTX). BTX was injected into dystonic muscles mostly into sternocleidomastoid then--trapezius, and splenius capitis muscle. Improvement (excellent, good and fair) was achieved in 40 patients (62%). Lack of information about 6 patient (9%). Injections were repeated every 3-4 months and in several cases even 1-2 during the year. After several injections atrophy and denervation potentials in EMG were observed in the majority of injected muscles. Neurotic syndromes coexisting with dystonia had worsening influence on therapeutic effects. Adverse events were observed in 5 cases. Treatment with BTX is very simple, easy, harmless and can be administered in outpatients. PMID- 10463250 TI - [Electroencephalographic studies of family members of patients with juvenile myoclonic epilepsy]. AB - Juvenile myoclonic epilepsy (JME) is a common idiopathic generalized epilepsy (IGE); it has a clinical and probably a strong genetic relation to the other IGE forms. Generalized spike/polyspike-wave discharges (SW/PSW) are typical of all IGEs. The aim of our study was to determine the incidence of epilepsy and SW/PSW in EEG of family members of 12 JME patients. 35 first degree relatives aged over 15 years were examined. 40 min EEG with 5 min HV were recorded. IGE was diagnosed in 3 (8.6%) persons: JME in 2 and childhood absence epilepsy (CAE) in 1 person. Six more relatives (17.1%) had typical SW/PSW traits in EEG. Thus the IGE features were found in 9 (25.7%) individuals--members of 7 out of 12 families (58%). EEG of 7 other relatives (20%) revealed non-specific episodic diffuse or focal abnormalities. The above results reveal higher incidence of different kinds of ICEs and typical EEG traits in families of JME patients. This findings confirm familial susceptibility to IGE and may be helpful in genetical counselling. PMID- 10463251 TI - [Controlled study of iprazochrome effectiveness (Divascan) in prophylactic treatment of migraine]. AB - The study was designed as double blind, placebo controlled. The patients were treated with 15 mg of iprazochrome daily in three equal doses for eight weeks, or -with equal amount of placebo tablets. The effectiveness was calculated with the use of Migraine Score (MS) by Couch et. al. 44 patients completed the study. In 21 the therapy was positive: 16 out of them were treated with iprazochrome, 5- with placebo. In 23 patients the treatment was negative: 19 out of them took placebo, 4--iprazochrome. Statistical analysis showed significant influence (chi 2 test: p < 0.001; Youle coeff. = 0.88). In iprazochrome group mean decrease of MS was significant after treatment (p < 0.01), but not significantly changed in the placebo group. According to our results iprazochrome was found effective in the prophylaxis of migraine. PMID- 10463252 TI - [Lumbar disk herniation treated by microsurgery]. AB - Microsurgical technique was applied in 46 cases of lumbar disc herniation in the years 1993-1994. The group comprised 17 women and 29 men aged 17-46 years (mean age 39.54 +/- 8.72). Sciatic pain was present in 39 (85%) cases, in the remaining ones low back pain was the main symptom. Neurological deficit signs were noted in 21 (46%) cases. The authors discuss the indications to microdiscectomy, the microsurgical technique and outcomes. Microdiscectomy was carried out in cases selected strictly according to the result of neurological examination and radiological findings X-ray of lumbosacral region in 46 cases (100%), myelography in 11 (24%), CT in 4 (9%) and MRI in 31 (67%). MRI served for evaluation of signal intensity, site and site of disc hernia. In 31 cases the operation was done in patients with recognized unilateral and one-level disc hernia. Full remission of complaints was noted in 45 patients during follow-up over 2 years. PMID- 10463253 TI - [Prognostic factors in patients with intracerebral hematoma caused by ruptured middle cerebral artery aneurysm]. AB - In a retrospective study, the authors analysed surgical outcomes in patients after intracerebral haemorrhage (ICH) as a result of ruptured middle cerebral artery (MCA) aneurysm. Between January 1989 to June 1997, 836 patients with ruptured aneurysm were admitted. Of these 207 (24.8%) patients had MCA aneurysm. Sixty-seven patients (32.4%) with MCA aneurysm had ICH. The types of ICH were classified into three groups according to CT findings on admission: A) temporal ICH (with or without a minor SAH); B) intrasylvian haematoma (with or without a minor SAH); C) ICH with diffuse SAH (SAH with cisternal clots on the side contralateral to the haematoma). The outcome was assessed according to the Glasgow Outcome Scale. Each patient was classified as having made either a good recovery including moderate disability (a favourable outcome) or a poor recovery including severe disability, vegetative state, or death (an unfavourable outcome). Overall, 31 patients (46.3%) had a favorable outcome (good recovery in 12 (17.9%) cases and moderate disability in 19 (28.4%) cases), and 36 patients (53.7%) had an unfavourable outcome (18 (26.9%) suffered severe disability, 4 (5.9%) remained in vegetative state, and 14 (20.9%) died. A temporal ICH occurred significantly more often in patients with favourable outcomes (67.7%) (p < 0.01). In patients with favourable outcomes the incidence of Grade I and II was higher (51.6%) than that in patients with unfavorable outcomes (19.4%) (p < 0.025). Surgical complications were significantly higher in patients with unfavourable outcomes (52.8%; p < 0.01). Patients who developed more than 25 ml of ICH had significantly worse outcomes (p < 0.05). Factors that could be used to predict a favourable outcome include temporal ICH, WFNS Grade I or II, absence of a surgical and postoperative complication, and a haematoma volume less than 25 ml. PMID- 10463254 TI - [Metastatic tumors in vertebral canal]. AB - Medical records of patients suffering from spinal cord or cauda equina compression secondary to metastatic tumour of the spinal canal were reviewed. Out of 55 patients treated in the Department of Neurosurgery in Lublin throughout 1976-1996 for metastatic disease of the spinal canal, forty were operated on. Clinical symptoms, methods of treatment and course of disease in this last group of patients were analysed. It showed that the great majority of patients are admitted to the neurosurgical department with severe neurological deficits. Eighty percent of patients (n = 40) presented with deterioration of gait and 67.5% had retention of urine, 72.5% had sphincter disturbances. The series comprised patients with carcinoma of the lung (22.5%), kidney (7.5%), prostate (7.5%), neoplastic disease of the lymphatic system (12.5%) and other malignant neoplasm accounting for 62.5%. As a general rule, decompressive laminectomy was carried out. Surgery was usually performed as an emergency in order to arrest progression of neurological deficits and to prevent irreversible damage to the roots and spinal cord. The tumours were usually extradural, located in the lower part of the thoracic spine and invading surrounding tissues, including bone. Histological diagnosis of spinal tumour as a metastasis preceded diagnosis of a primary tumour, but in 1/3 of cases histology could not indicate of the primary malignancy. There are substantial differences in the clinical course of a metastasis in the thoracic and lumbosacral spine. Own experience indicates that surgery of spinal metastases in patients with severe neurological deficits has a very limited effectiveness. PMID- 10463255 TI - [The use of echocardiographic studies in neurology]. AB - Transoesophageal echocardiography (TEE) has become a new valuable method complementary to transthoracic echocardiography (TTE), thus markedly increasing the diagnostic accuracy in cases of cardiac and great vessels pathology. Importance of TEE in patient care and therapy in modern cardiology is emphasized. Indications, contraindications, complications as well as advantages and limitations of the technique are discussed. The diagnostic value of TEE is compared to TTE. TEE allows to reveal other sources of cerebral embolism, such as paradoxical embolus, aortic embolism or left atrial thrombus. PMID- 10463256 TI - [The role of nitrogen oxide in migraine attack]. AB - Views are surveyed on the role of nitrogen oxide in the pathogenesis of migraine attacks. There is some evidence that molecule plays an important role in these attacks through vasodilation in the central nervous system. In the process of nitrogen oxide release glutaminergic receptors of NMDA type participate modulating the activity of serotoninergic receptors. Increased synthesis of nitrogen oxide can follow spreading cortical ischaemia, histamine release from mast cells in the meninges and also through activation of the trigeminovascular system. The hypothesis of active role of NO in the pathogenesis of migraine attacks is supported by the analysis of the mechanism of action of drugs effective in the treatment of that disease. PMID- 10463257 TI - [Amyotrophic lateral sclerosis]. AB - The literature on amyotrophic lateral sclerosis from the years 1995/96 is reviewed including the topics of the World Congress of Neurology in Buenos Aires in September 1997. The problems of a) changed criteria of diagnosis and course of SLA after the time of the first definitions by classical neurological authors, b) the concepts concerning the aetiology and mechanisms of the disease, c) the present studies on drugs and their tentative clinical applications are discussed. PMID- 10463258 TI - [Empty sella syndrome]. AB - The authors present two middle-aged female patients with empty sella revealed at imaging studies (CT, MRI). Their main complaint was severe fronto-parietal and fronto-temporal headache. Physical examination showed obesity, hypertension, and local hypersensitivity on deep palpation and percussion in the above-mentioned regions, in both cases. Endocrine function of pituitary gland, visual fields and fundi were normal as was EEG. The CSF composition and pressure also showed no abnormalities. The diagnostic and therapeutic problems of empty sella syndrome are discussed. PMID- 10463259 TI - [Temporal arteritis as a cause of a terrible headache]. AB - Two patients with temporal arteritis are reported in whom terrible headache was the predominant symptom of the disease. Both of them improved after prednisone 60 mg daily in the initial dose. The diagnosis and the treatment of giant cell arteritis are discussed. PMID- 10463260 TI - [Diagnostic problems in vertebral body hemangioma]. AB - Vertebral haemangioma is a slowly growing benign tumour arising from blood vessels. Haemangioma of the vertebrae is usually asymptomatic. Neurological symptoms occur when haemangioma causes compression of nerve roots or spinal cord. In this paper a case of a 59 year-old woman with, vertebral body L2 haemangioma is reported. The signs and symptoms, modern imaging techniques (CT, MRI), differential diagnosis and treatment were discussed. PMID- 10463261 TI - [A case of congenital fissure of C2 and C3 arches with compression of cervical spinal cord]. AB - A 41-years old male patient is reported who had spastic paresis of all extremities, and evidence of peripheral motor neuron damage. He had been treated with steroids for 10 years after erroneous diagnosis of multiple sclerosis. The final diagnosis was a congenital fissure of C2 and C3 arches with spinal cord compression in cervical part. After operation some improvement of the neurological status was obtained. PMID- 10463262 TI - [Subacute sensory neuronopathy with signs of involvement of anterior horn cells in a patient with small cell lung carcinoma (case report)]. AB - A 55 years old woman with small-cell lung carcinoma is described. Ten months after the diagnosis was established, subacute sensory neuronopathy with the signs of involvement of anterior horn cells (confirmed by EMG exam) occurred. Since neurological symptoms appeared at the time when anti-cancer treatment was ceased, the diagnosis of paraneoplastic lesion peripheral nervous system was established. Biopsy of sural nerve obtained 3 months after the onset of neurological signs showed nearly complete loss of normal looking myelinated fibers due to the process of axonal degeneration with relatively better preserved unmyelinated fibers. The patient died after 1 year and 2 months from the beginning of the disease because of metastatic tumours in the brain. PMID- 10463263 TI - [Anorexia as a symptom of cerebral aneurysm in a 14-year-old boy]. AB - The authors present a very rare case of a child with anterior communicating artery aneurysm and symptoms of anorexia nervosa. Because of coexistence of subarachnoid haemorrhage and head trauma false diagnosis of temporal and frontal lobe contusion was initially established. Headaches, anorexia and cachexia occurred with aneurysm enlargement. Computed tomography and cerebral angiography allowed to indicate operation. After successful surgery all symptoms disappeared. PMID- 10463264 TI - [Letter to the editor on classification and terminology of paraneoplastic syndrome of peripheral nervous system in small cell carcinoma of the lungs]. PMID- 10463265 TI - [Letter regarding the paper by D. Milewska, W. Swiderski, K. Owczarek: "Hospitalization for nervous system tumors in Poland in the period 1979-81 and 1986-88", Neurologia i neurochirurgia Polska, 1992, v. 26, 5, 627-36]. PMID- 10463266 TI - [Comparison of the effect of vincamine and nicergoline in the treatment of ischemic stroke]. PMID- 10463268 TI - Cystic neuroblastoma: emphasis on gene expression, morphology, and pathogenesis. AB - Cystic neuroblastoma (CN) is an unusual variant of neuroblastoma characterized by a grossly visible cyst(s) and almost always distinctive microcysts on light microscopy. Rarely, CN will appear solid grossly, but microcystification will be present. We examined the clinical, pathologic, and biologic features of 17 cases of CN. The majority of CN had been detected by prenatal ultrasound. The tumors were favorable stage, stroma-poor, but with low or intermediate mitotic karyorhectic indices and had favorable biologic markers reflected by aneuploidy and by an absence of N-myc amplification and chromosome 1p deletions. However, the high trk expression typically identified in good risk tumors was absent. Although the complete natural history of CN is not fully defined, our experience suggests that some tumors progress in size, whereas others may spontaneously regress or mature. The clinical outcome is excellent, as is expected in localized and stage 4S neuroblastoma in infancy. PMID- 10463267 TI - Necrotizing enterocolitis of the newborn: pathogenetic concepts in perspective. PMID- 10463270 TI - Central nervous system malformations in a perinatal/neonatal autopsy series. AB - Congenital malformations of the central nervous system (CNS) are among the most common anomalies, but data on the incidence of CNS malformations in autopsy populations are scant. We examined 4122 autopsies between the years 1958 and 1995. There were 363 cases (8.8%) with CNS malformations; 235 were neonates and 128 stillborns. The overall gender ratio was 1:1, although more male neonates and more female stillborns had malformations. The body weights ranged from 24 to 6440 g. Neural tube defects were the most common types of malformations (45.5%) and included anencephaly, meningoencephalocele, meningocele/meningomyelocele, craniospinal rachischisis, and spina bifida occulta. The incidences of other malformations were: congenital hydrocephalus (12.4%), neuronal/glial proliferation disorders such as micro- and macrocephaly (8.8%), neuronal migration disorders (8.8%), prosencephalon growth disorders such as holoprosencephaly and arhinencephaly (8.5%), abnormalities of the midline structures such as agenesis of corpus callosum (4.1%), developmental cysts (3.3%), cerebellar malformations (3%), and vascular malformations (2%). Miscellaneous malformations (3%) consisted of acephalia in four cases with twin reversed arterial perfusion (TRAP), two cases of hydranencephaly, and four cases of rare degenerative and metabolic encephalopathies. PMID- 10463269 TI - Cyclopamine-induced holoprosencephaly and associated craniofacial malformations in the golden hamster: anatomic and molecular events. AB - Holoprosencephaly is a complex congenital malformation of the brain and is often associated with a spectrum of facial anomalies ranging from normocephaly or nondiagnostic changes to cleft lip/palate (premaxillary dysgenesis), cebocephaly, ethmocephaly, and cyclopia. The primary insult is thought to occur during gastrulation, when prechordal mesenchyme and overlying anterior neural plate undergo complex developmental interactions. Exposure to cyclopamine, a steroid isolated from the desert plant Veratrum californicum, causes holoprosencephaly in mammalian embryos. We have begun to study the pathogenesis of cyclopamine-induced holoprosencephaly and associated craniofacial anomalies in Syrian golden hamsters (Mesocricetus auratus). Embryos were exposed to a single maternal dose of cyclopamine during gastrulation on embryonic day (E) 7.0. By E13.0, 62% of fetuses showed craniofacial malformations, including premaxillary dysgenesis, ocular hypotelorism, and cebocephaly. Facial anomalies were associated with absence of the premaxilla and abnormalities of the midline cranial base, particularly the ethmoid and sphenoid bones. Histological sections from cyclopamine-treated embryos at earlier stages showed marked deficiency of cranial mesenchyme derived from the rostral neural crest. Expression of two transcription factors, HNF-3 beta and Hox-b5, which have been implicated in specification of rostral and caudal neural crest cells, respectively, were examined immunohistochemically. Treatment with cyclopamine caused a transient loss of HNF 3 beta immunoreactivity in prechordal mesenchyme, but had no effect on Hox-b5 expression. The findings suggest that an early event in the pathogenesis of cyclopamine-induced holoprosencephaly may be altered expression of selected proteins in the prechordal mesenchyme and floor plate with secondary impaired development of the adjacent neural plate and cranial neural crest. PMID- 10463271 TI - Cell proliferation in the developing human kidney. AB - In a previous study, utilizing antibodies to proliferating cell nuclear antigen (PCNA), we determined the proliferation index (PI) (percentage of PCNA-positive cells) of intrinsic renal cell populations in the normal adult and pediatric kidney. We have found that the PI in both adult and pediatric kidneys was very low (below 0.5 in all examined cell populations). In our present study, we investigated cell proliferation in the developing human kidney with an antibody to PCNA. Histologically normal kidneys were collected from 25 fetuses (spontaneous abortions and stillborns) ranging from 10 wk of gestation to term. Immature mesenchyme (blastema), immature early tubules, ampulla of ureteric bud, proximal tubules, Tamm-Horsfall protein (THP)-positive tubules, distal tubules, collecting ducts, and glomeruli were evaluated separately. The PI for each cell population was calculated. The PI of immature early tubules remains high (33-43) throughout embryonic life. The PI of blastemal cells is initially similarly high, but gradually decreases starting from the second trimester. The PI of THP positive tubules, distal tubules, collecting ducts, and glomeruli starts out relatively high (5.9, 8.6, 6.0, and 12.4, respectively) and decreases gradually as term approaches (1.8, 1.3, 1.2, and 1.4, respectively). Interestingly, as soon as proximal tubules become differentiated (appearance of light microscopic features of proximal tubular epithelium with TP lectin positive brush border), their PI becomes very low (below 1) irrespective of the age of the kidney. This is the first quantitative study to show changes of the PI in various renal cell populations during human nephrogenesis. These changes in the PI relate to the stage of differentiation of the developing nephron segments. PMID- 10463272 TI - Histiocytoid cardiomyopathy: three new cases and a review of the literature. AB - Histiocytoid cardiomyopathy (HC), a rare arrhythmogenic disorder, presents as difficult-to-control arrhythmias or sudden death in infants and children, particularly girls. Three cases are described with autopsy findings. In two cases, yellow-tan nodules were grossly visible in the myocardium; in the third case, no gross lesions were identified. Microscopic examination in all three cases revealed multiple, scattered clusters of histiocytoid myocytes which on ultrastructural examination were filled with abnormal mitochondria, scattered lipid droplets, and scanty myofibrils. These pathologic findings are similar to those previously described. The pathogenesis of this entity remains controversial. It was recently proposed that this disorder is X-linked dominant with the associated gene located in the region of Xp22. PMID- 10463273 TI - Medullary ray nodule of the kidney. AB - In this short report we describe three preterm infants who died within the first 8 months of life (corrected age). All presented similar histological features within the kidney, showing medullary ray nodules composed of tubules lined by low columnar epithelium with clear cytoplasm. The immunohistochemical profile of these lesions was identical and suggested origin from distal convoluted tubule or collecting duct. The aim of this report is to draw attention to this entity and to suggest a possible origin of these lesions. PMID- 10463274 TI - Cystic fibrosis with fibrosing colonopathy in the absence of pancreatic enzymes. AB - Fibrosing colonopathy, characterized by dense submucosal fibrosis in the large bowel, is a disorder associated with bowel dysfunction in patients with cystic fibrosis who receive pancreatic enzyme supplementation. Most commonly, patients present with a distended abdomen and abdominal pain. Radiographs frequently demonstrate colonic wall thickening and luminal narrowing. Here I describe a neonate with cystic fibrosis who presented with both clinical and histological features of fibrosing colonopathy who had not received pancreatic enzymes. This report expands our understanding of the pathogenesis of fibrosing colonopathy. PMID- 10463275 TI - National Wilms Tumor Study: an update for pathologists. AB - The National Wilms Tumor Study (NWTS)-5, begun in August 1995, incorporates some important new definitions and concepts that have critical importance in determining therapy and prognosis. The criteria for stage 1 were refined to accommodate an important subset of stage 1 Wilms tumors that are being managed by nephrectomy alone, without the use of adjuvant therapy. The distinction between stages 1 and 2 in the renal sinus is no longer defined by the hilar plane but by the presence or absence of venous or lymphatic invasion. Specific problems encountered in the interpretation of the renal sinus are discussed here. The topographical definition of focal anaplasia has proven to be of prognostic value and is incorporated into the NWTS-5 study design. PMID- 10463276 TI - Metabolic and molecular bases of Menkes disease and occipital horn syndrome. AB - Menkes disease and occipital horn syndrome (OHS) are related disorders of copper transport that involve abnormal neurodevelopment, connective tissue problems, and often premature death. Location of the gene responsible for these conditions on the X chromosome was indicated by pedigree analysis from the time of these syndromes' earliest descriptions. Characterization of an affected female with an X-autosomal translocation was used to identify the Menkes/OHS gene, which encodes a highly evolutionarily conserved, copper-transporting P-type ATPase. The gene normally is expressed in nearly all human tissues, and it localizes to the trans Golgi network of cells. However, in over 70% of Menkes and OHS patients studied, expression of this gene has been demonstrated to be abnormal. Major gene deletions detectable by Southern blotting account for 15-20% of patients, and an interesting spectrum of other mutations is evident among 58 families whose precise molecular defects have been reported as of this writing. The center region of the gene seems particularly prone to mutation, and those that influence mRNA processing and splicing appear to be relatively common. Further advances in understanding the molecular and cell biological mechanisms involved in normal copper transport may ultimately yield new and better approaches to the management of these disorders. PMID- 10463277 TI - Making a difference. PMID- 10463278 TI - Histopathologic approach to metabolic liver disease: Part 1. PMID- 10463279 TI - Epidemiology of intrathoracic petechial hemorrhages in sudden infant death syndrome. AB - The possible effects of a wide range of sociodemographic and environmental factors on the incidence and distribution of petechiae were investigated in 485 sudden infant death syndrome (SIDS) cases from the New Zealand Cot Death Study. The number (nil, few, many) of macroscopic petechial hemorrhages in the visceral pleura, capsule of thymus, and epicardium was recorded in 458 of 474 autopsied SIDS cases. Other information was obtained from parental interview and obstetric records. Univariate analysis showed highly significant relationships (P < or = 0.005) between the frequency of petechiae at one or more sites and socioeconomic status, parity, breast feeding, age at death, time of death, sleep position, and head covering at death and lesser but significant relationships (P < or = 0.05) with Maori ethnicity, birth weight, gestation, pacifier use, and bed sharing. After multivariate analysis, significant associations remained between increased frequencies of thymic petechiae and parity (P = 0.0001), age at death (P = 0.0003), Maori ethnicity (P = 0.0019), pacifier use (P = 0.0001), and head covering at death (P = 0.0032); between increased frequencies of epicardial petechiae and head covering at death (P = 0.008) and an estimated time of death between 00:00 and 05:59 h (P = 0.056); and between increased frequencies of pleural petechiae and maternal smoking (P = 0.058) and parity (P = 0.022). There was a decreased frequency of pleural petechiae in infants placed prone for their final sleep (P = 0.058). The distribution and frequency of petechiae are affected by environmental factors, including known risk factors for SIDS, but these factors occur inconsistently across the three sites. The findings imply differences in the pathogenesis at each site but do not provide consistent support for previous theories of causation of petechiae. PMID- 10463280 TI - Bcl-2 expression in Langerhans' cell histiocytosis. AB - Langerhans' cell histiocytosis (LCH) is an abnormal accumulation of dendritic histiocytes of unknown pathogenesis. It has recently been shown to be a clonal process. Bcl-2 is a proto-oncogene whose protein product is known to inhibit apoptosis. The overexpression of bcl-2 has been demonstrated in a number of neoplasms, presumably prolonging the survival of the neoplastic cells. We examined the expression of bcl-2 in normal Langerhans' cells in the skin and in LCH by immunohistochemistry for protein and in situ hybridization for mRNA to see if it could be implicated in the pathogenesis of this disorder. Additionally, we performed Southern analysis to determine if genomic rearrangement of the bcl-2 gene occurs in cases of LCH. Bcl-2 was not detected in normal skin Langerhans' cells. Eleven of thirteen cases of LCH demonstrated bcl-2 protein expression in the cytoplasm of the Langerhans' cells by immunohistochemistry, while 12 of 13 cases had evidence of bcl-2 mRNA by in situ hybridization. Southern analysis revealed a germline configuration of the bcl-2 gene in the five cases studied. These findings suggest that bcl-2 expression is present and up-regulated in pathologic Langerhans' cells, however, this overexpression does not appear to be due to genomic rearrangement. PMID- 10463281 TI - Fascin and the differential diagnosis of childhood histiocytic lesions. AB - This is a descriptive screening of 46 examples of childhood histiocytic lesions and some of their look-alikes using a monoclonal antibody, p55, to fascin. Fascin, an actin-bundling protein, identifies dendritic cells in the blood and in tissues. Our aim was to test the diagnostic utility of the antibody in various lesions at different sites and to see whether the staining patterns give insight into the cell types involved. Fascin intensely stained the cells of juvenile xanthogranulomas (JXG), Rosai-Dorfman lesions, and soft tissue dendrocytomas. Normal Langerhans' cells and the cells of Langerhans' cell histiocytosis were unreactive. Their lack of fascin staining may be relevant to fascin being maturation as well as lineage related. Epithelioid and palisading granulomas were unstained, though an example of Kikuchi lymphadenitis had large numbers of dendritic-type cells that stained strongly. A reticulohistiocytoma of the skin was also unstained and look-alike lesions, Spitz nevi, and mast cell lesions did not stain. Two of three large-cell lymphomas (both CD30+) also had fascin reactivity. Even though fascin is not specific to dendritic cells, staining other cell types as well (false positive), and not entirely sensitive, dendritic cells such as tissue Langerhans' cells are unstained (false negative), there seems to be a consistency of staining in childhood histiocytic lesions. This may be of diagnostic use when read in the context of the tissue differential diagnosis. Whether fascin can serve as a marker for the dendritic cell lineage, or at least for some phases of dendritic cell lifecycle, is not answered by this survey. PMID- 10463282 TI - Vimentin immunohistochemistry in human fetal brain: methods of standard incubation versus thermal intensification achieve different objectives. AB - Vimentin is transiently expressed in many cells of neuroectodermal origin in the fetal central nervous system and may be demonstrated by immunohistochemistry in paraffin sections. Traditional methods incubate antibodies with tissue sections at room temperature, but boiling the incubating solution in a microwave oven enhances antigen retrieval and greatly intensifies immunoreactivity. This study compares vimentin immunoreactivity using various dilutions of a commercial monoclonal antibody and various incubation conditions including room temperature, overnight at 4 degrees C, and after heating to boiling either in a microwave oven or on a hot plate. Fifteen fetal brains and spinal cords were examined, ranging in age from 8 to 24 wk gestation. Normal brains of four term or near-term neonates and two with hypoxic/ischemic lesions were also studied. Hot plate and microwave heating were equivalent. No differences in immunoreactivity could be attributed to gestational age per se. Boiling for 10 min in a microwave oven at a dilution of 1:200 is recommended for the enhanced demonstration of fetal histological structures such as radial glial fibers, but it is not recommended for studies of maturational gradients of cellular vimentin that differ at each gestational age, as in fetal ependymal cells or during the conversion of radial glia to mature astrocytes. For semiquantitative developmental studies, vimentin incubation at room temperature or overnight in the refrigerator is recommended at a dilution of 1:25 or 1:50 to avoid misinterpretation of apparent excessive staining after thermal intensification. Recommendations of commercial suppliers focus on the identification of neoplastic cells and must be modified to demonstrate subtle developmental changes in fetal tissues. PMID- 10463283 TI - Intraoral tumor of Chievitz in a child. AB - Juxtaoral organs known as organs of Chievitz are intramuscular embryonic structures found close to the angle of the mandible near the insertion of the pterygomandibular raphae. They are considered of neuroepithelial origin with no known function. We describe the first tumor of the organ of Chievitz which presented intraorally in a child. Immunohistochemically, the Chievitz nests showed positive reaction for vimentin, cytokeratins, and epithelial membrane antigen and ultrastructurally demonstrated cytoplasmic processes and intermediate filament bundles. These observations, together with light microscopic features, suggest that the epithelial nests of the organ of Chievitz are meningothelial rather than neuroepithelial. PMID- 10463284 TI - Unique desmoplastic cerebral tumor in a patient with complex partial seizures. AB - Primary brain tumors with prominent desmoplasia include the gliofibroma, desmoplastic infantile ganglioglioma (DIG), pleomorphic xanthoastrocytoma (PXA), and desmoplastic cerebral astrocytoma. In the present report, we describe unusual pathological appearances in two successive resections of a left temporal tumor in a patient with complex partial seizures. Both tumors showed focal astroglial and very prominent neuronal differentiation. In the first resection at age 11 years, the tumor showed only focal desmoplastic areas and prominent neuronal differentiation with bizarre atypical giant cells. In the second resection at age 14 years, the morphology was dramatically different. Now the tumor showed marked desmoplasia with tumor cells coexpressing neuronal and glial markers. Electron microscopy showed prominent neuronal differentiation in both resections and presence of basal lamina around tumor cells. Our case represents a unique example of an extraordinary degree of neuronal differentiation in a desmoplastic cerebral tumor. While cellular pleomorphism in a desmoplastic tumor traditionally suggests the diagnosis of PXA, we wish to underscore that many cells exhibiting marked cytologic atypia may in fact be of neuronal rather than of glial origin as in our case. PMID- 10463285 TI - Pathological evaluation of renal tumors in children: international society of pediatric oncology approach. AB - In the majority of European countries, children with renal tumors now enter the SIOP-93-01 Trial and Study. The objective of this Study is to refine methods of treatment especially in stage I patients. The role of institutional pathologists is important in this trial. There are new criteria for stages I and II, a new SIOP Working Classification of Renal Tumors of Childhood, and morphologic and prognostic similarities of pretreated and non-pretreated anaplastic cases. Specific problems encountered in assessing tumors treated with preoperative chemotherapy, administered to the majority of children over 6 months of age entering the SIOP Study, are discussed. The identification of a new low-risk group, the completely necrotic Wilms tumor, is outlined. PMID- 10463286 TI - On a historical note: Duchenne of Boulogne. PMID- 10463287 TI - Histopathologic approach to metabolic liver disease: Part 2. AB - In this, part 2 of the histopathologic approach to the diagnosis of metabolic disease of the liver, the steatotic, cirrhotic, and neoplastic groups are addressed. See the previous issue, Volume 1, Number 3, of Pediatric and Developmental Pathology for part 1 [1]. The perspective concludes with a tabulated assessment of the likelihood of diagnostic ascertainment. PMID- 10463288 TI - Primary malignant epithelial tumors of the liver in children: a study of DNA content and oncogene expression. AB - Primary malignant epithelial tumors of the liver (PMETL) are rare in the pediatric age group, and very little is known about their biology as compared with adult tumors. The prognostic value of the DNA contents measured by image analysis and expression of oncogene c-erb2 and tumor suppressor gene p53 were studied in 30 cases of PMETL in children, including 24 with hepatoblastomas (HB) and 6 with hepatocellular carcinomas (HCC). p53 overexpression was detected in 12 out of 26 cases (46.0%), or in 3 of 5 HCC and 9 of 21 HB cases. A relatively high concordance of staining was observed with the two antibodies used (clone DO7, Dako and clone DO1, Santa Cruz Biotechnology). c-erb-B2 did not yield the characteristic membrane staining in any of the 27 cases in which reliable staining was obtained. However, 1 out of 4 patients with HCC and 1 of 23 with HB revealed strong granular cytoplasmic staining in several neoplastic cells. Interestingly, these were two of the three aneuploid multiploid cases. DNA histograms of 13 out of 29 cases (54.8%) were classified as DNA aneuploid (5/6 HCC and 8/23 HB): nine were hyperdiploid, one was hypodiploid (1HB), and three were multiploid (2HB and 1HCC). In the HB group, DNA aneuploidy was strongly associated with embryonal histological areas, suggesting that a disturbance in the process of cell differentiation is associated with marked genetic aberrations. Only the group of HB was submitted to univariate analysis of survival by the Kaplan-Meier method for age (< 24 months vs. > or = 24 months), sex, preoperative chemotherapy (yes vs. no), residual disease (metastasis, and/or unresectable tumor), p53 expression by immunohistochemistry (positive vs. negative), and DNA ploidy (diploid vs. aneuploid). Only residual disease at the time of diagnosis (P < 0.017) and preoperative chemotherapy (0.030) were found to be negatively correlated with biological behavior, estimated as overall survival. DNA aneuploidy tumors (P < 0.125) and male patients (P = 0.123) showed a trend toward a more aggressive clinical behavior, although the difference was not statistically significant. Combining DNA ploidy and residual disease, patients were categorized into three groups: group I, patients with no adverse prognostic factors, i.e., diploid tumors without residual disease; group II, patients with only one adverse prognostic factor, i.e., aneuploid tumor or residual disease; and group III, patients with both adverse factors, aneuploid tumors and residual disease at time of diagnosis. A log-rank test comparing the three survival curves showed a statistically significant difference between them (P < 0.003). Although the series of cases is small, the results of this study highlight the importance of including DNA ploidy in the protocols designed for HB in children by international cooperative groups. PMID- 10463290 TI - Variation in major pulmonary fissures: incidence in fetal postmortem examinations and a review of significant extrapulmonary structural abnormalities in sixty cases. AB - A retrospective study of 1513 fetal postmortem examination reports from 1967 to 1996 yielded 35 cases with anomalous major pulmonary fissures (2.3%), to which a further 25 cases were added from fetal postmortem examinations performed between 1929 and 1966. Of 60 cases with anomalous fissures, 43 had an absent right horizontal fissure and 8 had a supernumerary left horizontal fissure; the remaining 9 showed various patterns in which one lung lacked major fissures. Histological examination in 29 cases did not reveal any additional pulmonary abnormality and pulmonary maturity was appropriate for gestational age. Additional malformations were present in 40 cases and these were frequently multiple, the most common being central nervous system, cardiovascular, and genitourinary system defects; notable heart malformations (10 cases); hydrocephalus (5 cases); and cystic renal dysplasia (4 cases). Chromosomal abnormalities were demonstrated in six cases although this figure does not reflect their prevalence, as many cases predate the availability of karyotyping. In seven cases, including three with polysplenia syndrome and one with situs inversus totalis, there was evidence of an underlying abnormality of left-right asymmetry, and in 13 cases there was documented pulmonary hypoplasia. PMID- 10463291 TI - Melanotic neuroectodermal tumor of infancy: a molecular genetic study. AB - Melanotic neuroectodermal tumor of infancy is a rare but well-recognized entity in pediatric pathology. However, the relationship of this tumor to other pediatric small cell tumors with neuroectodermal features (such as neuroblastoma, Ewing sarcoma/peripheral primitive neuroectodermal tumor, and desmoplastic small round cell tumor) is undetermined. Molecular genetic studies of melanotic neuroectodermal tumor of infancy have not been reported. We studied three typical cases of melanotic neuroectodermal tumor of infancy in an attempt to link this tumor to other small cell tumors with well-characterized molecular genetic changes. Tests performed included: detection of MYCN gene amplification and deletion of 1p (all 3 cases), and presence of the t(11;22)(q24;q12) and the t(11;22)(p13;q12) translocations (2 of 3 cases). None of these tests yielded positive results. Thus, there is no genetic basis at present to link melanotic neuroectodermal tumor of infancy to neuroblastoma, Ewing sarcoma/peripheral primitive neuroectodermal tumor, or desmoplastic small round cell tumor. PMID- 10463289 TI - Cloacal dysgenesis sequence: observations in four patients including three fetuses of second trimester gestation. AB - Cloacal dysgenesis is a rare malformation sequence. We studied cloacal dysgenesis in four fetuses, including three at 18-22 weeks of gestation. All four fetuses showed a smooth perineum with absence of anal, urethral, and/or vaginal openings. The urinary bladder was dilated in 3 cases. The labia majora and minora were absent in the two female fetuses; the scrotum and penis were absent in one male fetus and hypoplastic in the second. The kidneys were either absent (1 case), dysplastic (1 case), or hydronephrotic (1 case). Normal kidneys were seen in a fetus of 20-22 weeks gestation in whom the urinary obstruction was alleviated by a vesicocolonic fistula. This fetus did not have pulmonary hypoplasia. Severe renal anomaly and pulmonary hypoplasia are the limiting factors for the survival of infants born with cloacal dysgenesis. The unique observation of normal kidneys and lungs in one of our fetuses, despite anhydramnios, suggests that the effect of oligohydramnios on lung development may be limited early in gestation, at least up to 20-22 weeks. It may also indirectly support the theory that there are factors other than oligohydramnios that interfere with early lung development, such as reduced production of a pulmonary growth factor by the kidney or reduced proline production by malformed kidneys, that may cause decreased collagen formation and result in hypoplastic lung mesenchyme. PMID- 10463292 TI - Deep granuloma annulare (pseudorheumatoid nodule) in children: clinicopathologic study of 35 cases. AB - Deep granuloma annulare (DGA) is one of several lesions of skin and superficial soft tissues whose histologic character is a palisading granuloma with a small central focus of necrosis or necrobiosis. Unlike the other palisading necrobiotic lesions, DGA has a predilection for children in the first 5 to 6 years of life. A painless subcutaneous nodule(s) in the lower anterior tibial region or foot and the scalp, typically in the occiput, was the most common presenting feature in this study of 35 cases. Additional or recurrent lesions were reported in approximately 70% of cases with clinical follow-up. All lesions showed the presence of necrobiosis; however, one of the characteristic features was the multinodular character of the predominantly mononuclear cellular aggregates. The presence of vascular spaces at the periphery of the nodular profiles served as a clue to the diagnosis of DGA. The palisading arrangement of the mononuclear cells was evident only in those foci with central necrobiosis. A histiocytic disorder or fibrohistiocytic process was a common consideration in the differential diagnosis, especially in those cases with less apparent foci of necrosis. Palisading histiocytes with prominent eosinophilic cytoplasm and some nuclear atypism were problematic with regard to possible epithelioid sarcoma. Our study failed to identify any underlying or predisposing factors in the development of DGA. Despite the fact that DGA is a well-documented lesion in children, it occurs sufficiently infrequently that it is often not considered clinically when it presents as a subcutaneous mass or masses in a child. Its recognition by the pathologist is especially important as the occurrence of additional lesions in a high proportion of children can be anticipated without undue concern. PMID- 10463293 TI - Detection of mycobacterial infections using the Dieterle stain. AB - Retrospective review comparing the modified Dieterle stain with standard acid fast stains was performed on seven surgical pathology cases that contained culture-positive mycobacteria infections. Tissues examined comprised cervical and submandibular lymph nodes and soft tissues of the face and chest. Modified Dieterle staining was performed on paraffin-embedded tissue sections, and the results were compared with those of hematoxylineosin stains and auramine rhodamines and carbolfuchsin acid-fast stains. The acid-fast stain showed organisms in three of seven cases on initial review and five of seven cases on retrospective review; the auramine-rhodamine stain retrospectively revealed organisms in five cases. In contrast, the Dieterle stain showed organisms in all seven sections on retrospective examination. Dieterle stains revealed either beaded bacilli or nocardia-like filamentous organisms, sometimes with abundant granular debris possibly representing degenerative organisms. In three cases in which bacteria were readily apparent with the Dieterle stain, only rare organisms could be identified with the acid-fast stain. The modified Dieterle stain was more sensitive than the acid-fast stain in the identification of mycobacteria in pediatric specimens, and its use is recommended in cases with necrotizing granulomas. However, its specificity is limited by morphologic similarities of the organisms to those of nocardiosis and cat scratch disease. PMID- 10463294 TI - Cleidocranial dysplasia with neonatal death due to central nervous system injury in utero: case report and literature review. AB - Cleidocranial dysplasia (CCD), an uncommon disorder involving membranous bones, is rarely lethal in early life. The calvaria is defective and wormian bones are present. Abnormalities of the clavicles vary in severity from a minor unilateral defect to bilateral absence. This report concerns pre- and postmortem anatomical and radiological findings in a 15-day-old female neonate with CCD. Her postnatal course was characterized by seizures and recognition of hydrocephalus during the first day of life. The calvaria was hypoplastic with numerous wormian bones. A pseudofracture of the right clavicle was present. Hydrocephalus was present in the brachycephalic brain which had a severely thinned cerebral cortex. Hemosiderin in the ventricular lining and marked subependymal gliosis were interpreted as evidence of old intraventricular hemorrhage that had occurred in utero. A CCD-related condition, Yunis-Varon syndrome (YVS), is noted for early lethality and for developmental and secondary abnormalities of the central nervous system. The present case only partially matches the phenotype of YVS and might represent a part of a spectrum of phenotypic variants ranging from viable CCD to lethal YVS. PMID- 10463295 TI - Monoclonal gammopathy of the immunoglobulin A class in a two-year-old girl with ataxia telangiectasia. AB - Ataxia telangiectasia is an autosomal recessive neurologic disorder which is frequently associated with a deficiency of IgA immunoglobulin. We report an unusual case of monoclonal gammopathy of the IgA kappa type in a 2-year-old female patient newly diagnosed with ataxia telangiectasia. Quantitative analysis of the patient's immunoglobulins revealed a marked elevation in the IgA fraction with a value of 672 mg/dL (normal 14-123 mg/dL). The IgG and IgM fractions were normal. Serum protein electrophoresis showed a band of restricted mobility present in the gamma region, which was identified as a monoclonal IgA kappa immunoglobulin on immunofixation electrophoresis. This is the first case report of a patient with ataxia telangiectasia associated with an IgA monoclonal gammopathy. PMID- 10463296 TI - Congenital thyroid teratoma: a case with persistent neuroglial involvement of cervical lymph nodes. AB - A case of congenital thyroid teratoma with nodal spread is reported. Primary surgery was performed on a female infant on the 6th day of life. The thyroid mass was removed in toto, and an adjacent 1.2 cm lymph node was also removed. Histology showed solid and cystic teratoma with a variety of elements including prominent neurological tissue that was neuroblastoma-like in places. Residual compressed non-neoplastic thyroid tissue was identified in the subcapsular plane. The lymph node was largely replaced by neuroglial tissue that was cellular in some areas and showed intrasinusoidal growth and some mitotic activity. Recurrent cervical lymphadenopathy gradually developed, commencing a few months after surgery. Excision of cervical nodes was undertaken at 9 months of age. About 13 nodes up to 2 cm in diameter were excised. Most of the specimens consisted of reactive lymph nodes, but in three of the smaller nodes, there were subcapsular and sinusoidal masses of focally cellular neuroglial tissue, again with occasional mitoses. This tissue stained strongly for glial fibrillary acidic protein, in addition to expressing neural markers. The lymph node "deposits" were interpreted as "displaced" lesional tissue rather than metastases in the usual, aggressive sense. The girl remains well at 5 years of age. PMID- 10463297 TI - Pediatric germ cell tumors: protocol update for pathologists. AB - The therapy for pediatric germ cell tumors has historically been widely variable and institution dependent. The efforts to provide consistent, biology-driven therapy through Intergroup protocols has been a relatively recent phenomenon that has provided the framework for future protocol designs. The first Intergroup protocols confirmed that stage I malignant testicular germ cell tumors and immature teratomas at all sites in children could be treated with surgery alone followed by close observation. Future protocols currently in the planning stages may extend this low-risk category to stage I ovarian germ cell tumors of all histologic types. Low-stage extragonadal germ cell tumors may be placed in a new intermediate risk category. Particular pathologic issues that were raised and reported during the first protocols and that may impact on future protocol design include the presence and size of foci of endodermal sinus tumors within low-stage immature teratomas at all sites. Accurate staging will grow more critical in future protocols. The ability to recruit international cooperative groups will determine the success of chemotherapy tailored to specific subgroups that now must be lumped for statistical purposes. Lastly, companion biologic studies will be critical to defining the different subtypes of germ cell tumors and to determining predictors of biologic behavior. PMID- 10463298 TI - Laser-Doppler Flowmetry for microcirculation monitoring. Introduction. PMID- 10463299 TI - Higher-order spectral analysis in laser-Doppler flowmetry signal processing. AB - This paper presents an application of higher order statistics and spectra to the problem of laser-Doppler Flowmetry (LDF) for microcirculation monitoring. The proposed signal processing technique computes the bispectra of LDF signals after appropriate pre-processing and extracts features which can be used for classification of the signals to a normal or patient category. Bispectra are defined in terms of the third-order moments or cumulants of signals and are shown to contain valuable information for the above classification of LDF signals. Experimental studies, including (a) a set of 17 normal subjects and 69 patients with Raynaud's phenomenon and (b) 50 LDF-signals separated to four groups from patients that have different pathologic characteristics, are described and results are presented which illustrate the performance of the proposed approach when applied to the LDF signals. PMID- 10463300 TI - Time-variant spectral analysis of LDF signals on the basis of multivariate autoregressive modelling. AB - Time-variant AR-modelling was used to analyse the non-stationary properties of LDF signals during provocation tests. The procedure for the estimation of time varying AR parameters based on Kalman filtering is presented. The estimates can be used to calculate instantaneous measures, such as peak frequency, spectral band power and coherence. The resulting course of instantaneous peak frequency of a sinusoidal signal with frequency jump was compared to similar parameters derived from short-time FFT and Hilbert transformation. Univariate time-variant spectral analysis was used to investigate LDF measurements in patients with Raynaud's phenomena. The experimental protocol was splitted into phases of different room temperature. The results demonstrate time-dependent variations of spectral components (amplitude and frequency). By means of time-variant coherence analysis of LDF and diameter measurements of vessels in a hamster skin fold the existence of a main rhythm around 0.1 Hz in the LDF signals which is related to vasomotion is shown. PMID- 10463301 TI - Different flowmotion patterns in healthy controls and patients with Raynaud's phenomenon. AB - Flowmotion was characterized in healthy controls and 61 Raynaud's phenomenon (RP) patients by spectral analysis of laser-Doppler perfusion monitoring (LDPM) tracings. Healthy subjects flowmotion patterns revealed a main frequency of 3 cycles per min (cpm) with another low frequency and heart rate synchronous components. A first group of RP patients presented a low frequency and heart rate frequency component but no significant difference in blood flow. The second group presented the predominating heart rate related frequency with low microvascular perfusion. The third group presented a flowmotion pattern with overlapping of heart rate and low frequency components. Patients with primary and secondary RP show specific changes in flowmotion, probably related to increased sympathetic nervous activity or vessel wall alterations causing disappearance of arteriolar tone and impairment of microvascular perfusion. The group of patients with overlapping frequency components presents an intermediate flowmotion pattern indicating a different grade of alterations in microvasculature. PMID- 10463302 TI - A method for digital signal processing based laser-Doppler flowmetry. AB - Non-invasive measuring of blood perfusion of tissue is important for establishing the influence of vascular diseases on microcirculation. A signal processing method for laser-Doppler flowmetry is presented in this article. The method is based on the digital filtering. A couple of filters are suggested. One of them is a digital passband filter, second - a digital filter transforming spectrum P(f) of the input signal to fP(f) spectrum. Performance of the suggested method is investigated in comparison with behaviour of the PERIMED instrument as well as with applications based on DFT. PMID- 10463303 TI - Extraction of principal components from biosignals by neural net. AB - This contribution gives the information on a useful application of principal component analysis (PCA) in the field of electroencephalogram (EEG) and laser Doppler signal processing. The principal components are estimated by a neural network (NN) approach. PMID- 10463304 TI - Principles and practice of the laser-Doppler perfusion technique. AB - This paper reviews the development and use of laser-Doppler perfusion monitors and imagers over the past two decades. The enormous interest in microvascular blood perfusion coupled with the 'ease of use' of the technique has led to 1500+ publications citing its use. However, useful results can only be achieved with an understanding of the basic principles of the instrumentation and its application in the various clinical disciplines. The basic theoretical background is explored and definitions of blood perfusion and laser-Doppler perfusion are established. The calibration method is then described together with potential routes to standardisation. A guide to the limitations in application of the technique gives the user a clear indication of what can be achieved in new studies as well as possible inadequacy in some published investigations. PMID- 10463305 TI - Spectral analysis of laser-Doppler perfusion signal measured during thermal test. AB - The method of spectral analysis of laser-Doppler perfusion signal measured during thermal test is proposed. During three 20 min phases with 40, 5, and 40 degrees C of thermal test laser-Doppler perfusion signal was recorded. For each signal phase frequency spectra were calculated using the FFT method. Quantitative parameter Spectral Factor for results evaluation was proposed. In total 94 patients were measured: 69 with Raynaud's phenomenon and 25 normal subjects. Additionally in 18 Raynaud's patients the influence of Nifedipine was studied. Results suggest that proposed parameter is able to differentiate between Raynaud's patients and normal subjects and that is useful for evaluation of Nifedipine effectiveness. However, further studies are needed to improve the method to differentiate between primary and secondary Raynaud's patients. PMID- 10463306 TI - Doppler Monte Carlo simulations of light scattering in tissue to support laser Doppler perfusion measurements. AB - Doppler Monte Carlo (DMC) simulations of the transport of light through turbid media, e.g., tissue, can be used to predict or to interpret measurements of the blood perfusion of tissue by laser-Doppler perfusion flowmetry. We describe the physical and mathematical background of Doppler Monte Carlo calculations, and present some comparisons with measurements, performed with experimental flow models, built to mimic skin tissue characteristics and for the calibration of perfusion instruments,which are important goals of the Biomed-I concerted action. The measurements deal with coherence effects. PMID- 10463307 TI - Tissue motion--a disturbance in the laser-Doppler blood flow signal? AB - The tissue matrix in the human body is continuously in motion driven by forces generated by the circulatory and respiratory systems. The linear velocities recorded can be as high as 300 microm/s. The question raised: is the motion of tissue matrix generating an error signal in laser-Doppler (LD) recordings that is misinterpreted as solely a blood flow signal. The problem is studied by using a physical model demonstrating the contribution from a flow channel and, in addition, from a linear motion of the flow tube. Measurements of skin motion in the forearm have been performed. The corresponding spectra have been compared with blood flow spectra recorded with the LD method at the same sites. The two types of spectra overlap each other indicating that motion induced signals can be interpreted as a flow signal. The "blood flow" contribution from tissue motion varies strongly with the site studied. PMID- 10463308 TI - Dynamic characteristics of laser-Doppler flux in normal individuals and patients with Raynaud's phenomenon before and after treatment with nifedipine under different thermal conditions. AB - The study was aimed at choosing an appropriate characteristic of laser-Doppler flux (LDF) data for (1) distinguishing patients with Raynaud's disease from normal controls and (2) evaluating the effect of nifedipine under different thermal conditions. We checked the reliability of three characteristics of nonlinear dynamics as statistical dimension Ds, correlation dimension D2 and power-law index PLI. Their values depended heavily on the thermal condition. The most reliable characteristics that enabled us to distinguish the patients from normal controls and the effect of nifedipine under definite thermal condition proved to be Ds and PLI. The latter is simple for computation and is thus recommendable for clinical practice. Ds and PLI were higher in patients with Raynaud's compared to normal controls and diminished during the transitions from low to high temperature. However, the characteristics used were unable to distinguish significantly Raynaud's I from Raynaud's II patients. PMID- 10463309 TI - Dynamic characteristics of laser-Doppler flux data. AB - Methods for tracking the dynamics of the blood flow microcirculation obtained by laser-Doppler flowmetry (LDF) technique are described. It was shown that LDF signals have complex dynamics. It was mainly characterized by fractal structures and chaos, though multiperiodic, trend-like and stochastic components were also established. Procedures for (i) describing the dynamic structure and (ii) tracking the dynamic changes in time of LDF data are proposed. Examples illustrating the efficiency of these procedures are given using both simulated and LDF data collected in experiments with reactive hyperemia. Irrespective of the universality of the methods, the procedures should be specified according to the problem-oriented clinical and experimental studies. PMID- 10463310 TI - Processing of laser-Doppler signals from free flaps. AB - As conventional graphical laser-Doppler flowmetric (LDF) registrations from free flaps are difficult to interpret we explored the use of refined computerized signal processing to enhance the reliability of the blood flow supervision postoperatively. From eleven free flaps LDF data were collected using a software programme and a personal computer for analysis. Findings were compared with the clinical outcome. Nine flaps healed whereas one had wound problems and one suffered a partial necrosis. From the nine uneventful flaps, a peak within the range of frequencies from 0.04 to 0.23 Hz was seen. In the remaining two, such a low frequency peak could hardly be observed. Frequency analysis using computerized processing of LDF signals thus has the capacity to demonstrate the status of the flap perfusion. The slow wave vasomotion component seems to be of particular importance. Other frequency components warrant further investigation. A custom made monitoring device would be of great clinical value. PMID- 10463311 TI - Linear and nonlinear analysis of blood flow in healthy subjects and in subjects with Raynaud's phenomenon. AB - The paper presents analyses of the dynamics contained in the blood flow signals measured on healthy subjects and on subjects with primary Raynaud's phenomenon. Different signal processing methods are presented and discussed. The dynamics was evaluated in the time and frequency domains and in phase space. Additionally, changes in the basal value during temperature provocation were studied using multiresolution analysis. The analyses demonstrate differences between the blood flow dynamics in healthy subjects and subjects with Raynaud's phenomenon. Moreover, the observed decrease in the amplitude of oscillation in regions approximately 0.04 Hz and approximately 0.1 Hz suggests an impairment in the neurogenic and the myogenic regulation of the blood flow. The administration of nifedipine in subjects with Raynaud's phenomenon results in an increase in the basal value and in the amplitude of the blood flow component oscillating with the heart rate. However, it does not restore the dynamics to that found in healthy subjects. PMID- 10463312 TI - Effects of prepulses and d-amphetamine on performance and event-related potential measures on an auditory discrimination task. AB - RATIONALE: Prepulse inhibition (PPI) of the startle reflex is a measure of sensorimotor gating, that is the processing of the startle stimulus (S2) is inhibited by the interfering processing of a closely preceding prepulse (S1). It has been demonstrated that PPI is disrupted in a variety of mental disorders and that several neurotransmitter systems, including dopamine, participate in the modulation of sensorimotor gating. Previous studies have also shown that a task relevant S1 enhances PPI in healthy subjects but not in schizophrenic patients. These findings indicate an influence of attentional processes on sensorimotor gating and an impairment of this modulation in schizophrenia. OBJECTIVE: Assuming a dopamine-mediated suppression of S1 processing as a mechanism of resource management and selective attention, which might be impaired in certain mental disorders, the present study investigated the effects of the indirect dopaminergic agonist d-amphetamine on prepulse-altered S2 discrimination and event related potentials (ERPs). METHODS: Twelve healthy volunteers were tested in a double-blind, placebo-controlled experimental design. Here, S2 is the target in a difficult Go/NoGo auditory discrimination task. RESULTS: Confirming our previous results, S2 processing is "accentuated" by a weak acoustic prepulse in healthy subjects, thus leading to a lower rate of errors of omission but also to more false alarms (i.e. a liberal response bias). This performance change correlated with a prepulse-induced increase in the amplitude of the P3 ERP towards non-targets ("prepulse-induced non-target positivity"; PINTP). In addition, the results of the present study show that under prepulse conditions amphetamine disrupts "S2 accentuation" associated with a dose-related reduction of the P2 component of the S1 response and a plasma level related reduction of PINTP. CONCLUSIONS: These data suggest an involuntary attentional shift towards S1 processing with increasing dopamine-release similar to that observed in patients with schizophrenia or OCD. It is concluded that sensory gating alters selective attention via dopaminergic modulation. PMID- 10463313 TI - Changes in sleep after acute and repeated administration of nicotine in the rat. AB - RATIONALE: Acetylcholine clearly plays a role in regulating sleep. This influence may involve nicotinic systems because several studies have demonstrated that nicotine treatment alters sleep. However, the literature that suggests an effect of nicotine treatment on sleep is contradictory, perhaps because different doses and routes of administration were used. OBJECTIVE: The studies reported here evaluated the effects of several doses of nicotine on REM sleep in the rat. METHODS: Male Wistar rats were prepared with a set of sleep recording electrodes and, following habituation to the test chamber, were used in one of three studies: a) a dose-response analysis of an acute dose of nicotine on REM sleep measured during the first 4 h after injection; b) a chronic treatment experiment; or c) a mecamylamine blockade experiment. RESULTS: Acute nicotine administration decreased REM sleep in a dose-dependent fashion; significant effects were observed following injection with the 0.5 and 1.0 mg/kg doses. A decrease in slow wave sleep and an increase in wakefulness were also observed. Mecamylamine by itself did not affect REM sleep, but it blocked the effects on sleep produced by nicotine when given 30 min before a 1 mg/kg dose of nicotine. Rats that had been injected once daily with a 0.1 mg/kg dose of nicotine showed an increase in REM sleep after the third injection, whereas rats that had been chronically treated with a higher dose (0.5 mg/kg) displayed a reduction in REM and total sleep time. CONCLUSION: These findings argue that the effects of both acute and chronic nicotine treatment on sleep are influenced by the dose of nicotine used. PMID- 10463314 TI - Midazolam effects on implicit and explicit memory processes in healthy subjects. AB - RATIONALE: Studying midazolam-induced amnesia offers an interesting approach to the organization of normal memory processes, since memory performance can be studied in the same subject in "on" and "off" drug conditions. OBJECTIVE: The present study investigated the effect of midazolam on skill learning. The task and the experimental design we used also allowed us to assess the effect of midazolam on priming and explicit memory. METHODS: Eighteen patients who underwent minor ear surgery and who were anaesthetized with intravenous midazolam, and 18 matched control subjects participated in a mirror reading task on 2 separate days. Patients were tested under midazolam on day 1 and without any medication on day 2. The mirror reading task was made of French words, some of which repeated across trial blocks, others being new. RESULTS: Patients under midazolam read new mirror written words faster with practice, which attested for intact skill learning. Moreover, they read repeated words faster than new words with practice, which was interpreted as reflecting intact priming abilities. These spared implicit memory capacities were observed along with severe explicit memory impairments. Learning for both new and repeated mirror written words on day 2 was similar in patients and in controls, which was interpreted as suggesting that the implicit learning that occurred on day 1 under midazolam was normal. CONCLUSIONS: The quality of skill learning, both in terms of speed of and lasting effect, was normal under midazolam in the task we used. In the context of the present task, midazolam offers an interesting, reversible model of amnesia. PMID- 10463315 TI - Repeated limited access to i.v. cocaine self-administration: conditioned autonomic rhythmicity illustrating "predictive homeostasis". AB - RATIONALE: Regular cocaine self-administration may act as a "Zeitgeber" in animals and humans, and physiological rhythms may peak at the time when the drug is usually self-administered. OBJECTIVE: To investigate whether or not daily sessions of cocaine self-administration for 2 weeks would synchronize autonomic rhythms to the time of daily access. METHODS: Two weeks after implant of a jugular catheter and telemetry sender, each rat's heart-rate (HR) and core temperature (Tc) rhythms were well entrained to the 12:12-h light:dark cycle (lights off: 0900 hours). For the next 2 weeks rats self-administered cocaine during daily sessions at 1300 hours. The circadian and ultradian oscillations of HR and Tc recorded during the 3 days after the last self-administration (SA) session were quantified by multi-oscillator cosinor analysis. RESULTS: The HR and Tc were elevated during the initial 0.25 mg cocaine infusions of the daily session. This initial tachycardia disappeared as cocaine continued to be self administered in accumulating amounts, while the locomotor activity remained elevated. After the last SA session the peak (acrophase) of the two cycles/day component in HR and Tc rhythms aligned to the time of day when cocaine self administration had previously been scheduled. The amplitude of the circadian and ultradian HR and Tc oscillations as well as their circadian acrophase were not impacted by repeated daily time-limited cocaine self-administration. CONCLUSIONS: Cocaine controls the behavior and autonomic functions of rats self-administering limited amounts of cocaine during regular daily sessions, and this controlling influence extends beyond the last daily access period. PMID- 10463316 TI - The interaction of cocaethylene and cocaine and of cocaethylene and alcohol on schedule-controlled responding in rats. AB - RATIONALE: Cocaethylene is a unique metabolite of cocaine, produced only in the presence of alcohol. This metabolite is pharmacologically, physiologically and behaviorally active. Further, it has been reported to interact pharmacokinetically with both cocaine and alcohol, an interaction that may mediate, in part, the interaction of cocaine and alcohol. Although cocaethylene has been shown to interact with both cocaine and alcohol, behavioral assessments of these interactions are limited. OBJECTIVES: To examine directly the behavioral interactions between cocaethylene and cocaine and between cocaethylene and alcohol, the present study assessed the effects produced by these combinations on schedule-controlled responding. METHODS: Rats were first administered cumulative doses of cocaethylene, cocaine and alcohol to assess their effects alone on responding. Following this, doses of cocaethylene were combined with cumulative doses of cocaine or alcohol. Additionally, doses of cocaine or alcohol were given in combination with cumulative doses of cocaethylene. RESULTS: When administered alone, cocaethylene, cocaine and alcohol produced dose-related decreases in responding. Further, cocaethylene shifted the dose-response functions for both cocaine and alcohol to the left and down, while cocaine and alcohol shifted the dose-response function for cocaethylene to the left and down. An isobolographic analysis revealed that these interactions were additive in nature. CONCLUSIONS: The present study suggests behavioral interactions between cocaethylene and cocaine and between cocaethylene and alcohol. The contribution of cocaethylene to the enhanced effects produced by the co-administration of cocaine and alcohol was discussed. PMID- 10463317 TI - Onset, magnitude and duration of opioid blockade produced by buprenorphine and naltrexone in humans. AB - RATIONALE: One therapeutic benefit of mu opioid agonist or antagonist maintenance is the resultant attenuation of the effects of illicit opioids. It is important to characterize the development and duration of opioid blockade produced by buprenorphine, a novel opioid dependence pharmacotherapy. OBJECTIVE: This study characterized the ability of buprenorphine to attenuate opioid effects during treatment initiation and discontinuation compared to naltrexone and placebo. METHODS: Opioid-experienced volunteers (n = 8) participated in this 10-week, inpatient, double-blind, within-subject, crossover study. Five randomized conditions [buprenorphine (2 and 8 mg, sublingually), naltrexone (25 and 100 mg, PO) and placebo] were each examined during a 2-week period; the test drug was given for 7 days followed by a 7-day placebo wash-out. Cumulative doses of hydromorphone (0, 2 and 4 mg, IM, 45 min apart) were administered thrice-weekly corresponding with treatment and wash-out days 1, 3, and 5; behavioral, physiological and pharmacokinetic measures were collected. RESULTS: Buprenorphine alone produced dose-related prototypic agonist effects during induction (i.e., positive mood, respiratory depression, miosis); tolerance developed only to the subjective effects. Buprenorphine 2 mg partially attenuated the effects of hydromorphone, while nearly complete attenuation was observed with 8 mg that lasted up to 72 h after discontinuation. Both naltrexone doses produced complete hydromorphone blockade after a single dose; blockade of the behavioral, but not physiological, effects persisted for 5 days after discontinuation of 100 mg. CONCLUSIONS: These data suggest that 2 mg buprenorphine is a sub-therapeutic maintenance dose, both buprenorphine 8 mg and naltrexone produce immediate and efficacious opioid blockade, and adequate protection against illicit opioids may be achieved with less-than-daily dosing. PMID- 10463318 TI - Platelet membrane fluidity individuals at risk for Alzheimer's disease: a comparison of results from fluorescence spectroscopy and electron spin resonance spectroscopy. AB - RATIONALE: Previous fluorescence studies employing 1,6-diphenyl-1,3,5-hexatriene (DPH) have revealed an increase in the fluidity of platelet membranes from individuals with Alzheimer's disease (AD) and their first-degree relatives. This biophysical alteration has been reported to be relatively specific for the hydrocarbon core of platelet membranes, where DPH preferentially localizes; this effect is not reflected by the fluorescent reporter triethylamino-DPH, which labels membranes at the lipid-aqueous interface. OBJECTIVE: The goal of this study was to explore the validity and reproducibility of these findings using an independent biophysical technique, electron spin resonance (ESR) spectroscopy. METHODS: Platelet membranes prepared from first-degree relatives of patients with AD were labeled with DPH, or the spin-labeled fatty acid probes 5-doxylstearate (5-DS) and 12-doxylstearate (12-DS). These spin labeled probes provide an index of structural order at the respective depths of their nitroxide moieties in the membrane. The resulting preparations were examined by fluorescence and ESR spectroscopy. RESULTS: Increased platelet membrane fluidity (PMF), as determined by the fluorescence anisotropy of DPH, was associated with only a modest reduction in the order parameter derived for 5-DS labeled membranes. In contrast, the mean order parameters derived from the paired samples labeled with 12-DS differed substantially from each other, and revealed decreased order (increased fluidity) in the hydrocarbon 12-C region where DPH preferentially localizes. CONCLUSIONS: These results provide an independent validation of the biophysical alterations of platelet membranes that are manifested by a subgroup of patients with AD and their first-degree relatives. PMID- 10463319 TI - The influences of age and caffeine on psychomotor and cognitive function. AB - RATIONALE: The response to caffeine is affected by a number of factors, including age. Older subjects may be more sensitive to the objective effects than younger but report fewer subjective effects. OBJECTIVE: This study assessed the influence of age on the effects of caffeine on a variety of psychomotor, cognitive and subjective tests. METHODS: Forty-eight healthy subjects, male and female, were recruited, 24 in the age range 20-25 and 24 in the range 50-65 years. All subjects were regular moderate caffeine drinkers and were not withdrawn from caffeine before entry to the study. A double-blind parallel group design was used with two groups of 12 subjects in each age range. One group in each age range received placebo and the other 250 mg caffeine B.P. A range of tests was used to assess psychomotor, cognitive and subjective functioning before and 1 h post treatment. RESULTS: Before treatment, young subjects generally performed better than older on psychomotor and cognitive tests. On the subjective tests, however, older subjects rated themselves as more alert and less tired than the younger ones. After placebo, performance and alertness improved in the younger group but declined in the older. After caffeine there were improvements in psychomotor performance and cognitive functioning in both groups, particularly in offsetting declining performance over time in the older subjects. It also produced subjective improvements in alertness. One factor to emerge was that on most assessments older subjects were better earlier in the day whereas in younger subjects performance did not show the same magnitude of decline throughout the day. CONCLUSIONS: Caffeine induced small but significant improvements in vigilance and psychomotor performance. PMID- 10463320 TI - Effects of itraconazole on the steady-state plasma concentrations of bromperidol and reduced bromperidol in schizophrenic patients. AB - RATIONALE: A previously reported pharmacokinetic interaction between bromperidol and carbamazepine, an inducer of cytochrome P450 (CYP) 3A4, suggests possible involvement of CYP3A4 in the metabolism of bromperidol. OBJECTIVE: We investigated pharmacokinetic interaction between bromperidol and itraconazole, a potent inhibitor of CYP3A4, to clarify the involvement of CYP3A4 in the metabolism of bromperidol and its reduced metabolite. METHODS: Itraconazole 200 mg/day for 7 days was coadministered to eight schizophrenic patients treated with a fixed dose of bromperidol 12 or 24 mg/day for at least 2 weeks. Blood samples were taken before and 1 week after itraconazole coadministration and 1 week after its discontinuation, together with clinical assessments using the Brief Psychiatric Rating Scale (BPRS) and the Udvalg for Kliniske Undersogelser (UKU) Side Effect Rating Scale. RESULTS: Plasma concentrations of bromperidol during itraconazole coadministration (16.7+/-4.9 ng/ml) were significantly higher (P<0.01) than before itraconazole coadministration (8.9+/-4.4 ng/ml) and 1 week after its discontinuation (9.9+/-4.3 ng/ml). Plasma concentrations of reduced bromperidol during itraconazole coadministration (3.6+/-2.9 ng/ml) were significantly higher (P<0.01) than before itraconazole coadministration (1.8+/ 1.3 ng/ml). No changes were observed in BPRS and UKU scores throughout the study. CONCLUSIONS: The pharmacokinetic interaction between bromperidol and itraconazole is probably due to the inhibitory effect of itraconazole on the metabolism of bromperidol. This study provides in vivo evidence of involvement of CYP3A4 in the metabolism of bromperidol and reduced bromperidol. PMID- 10463321 TI - Interactive effects of subanesthetic ketamine and haloperidol in healthy humans. AB - Ketamine is an N-methyl-D-aspartate (NMDA) receptor antagonist with prominent psychoactive effects in humans. This study evaluated whether the oral administration of haloperidol 5 mg would block the effects of an intravenous ketamine infusion (bolus of 0.26 mg/kg followed by 0.65 mg/kg per hour). Twenty healthy subjects completed 4 test days involving the oral administration of haloperidol or matched placebo 2 h prior to the intravenous infusion of ketamine or saline. Ketamine produced cognitive, behavioral, neuroendocrine, and physiologic effects in the healthy subjects that were similar to previous reports. Haloperidol pretreatment reduced impairments in executive cognitive functions produced by ketamine as measured by proverb interpretations and the Wisconsin Card Sorting Test. However, it failed to block the capacity of ketamine to produce psychosis, perceptual changes, negative symptoms, or euphoria in healthy subjects. These data outline an important, but functionally delineated modulation of ketamine effects by dopamine2 receptors and other sites of haloperidol action. PMID- 10463322 TI - Reinforcing and discriminative stimulus effects of the neuroactive steroids pregnanolone and Co 8-7071 in rhesus monkeys. AB - RATIONALE AND OBJECTIVES: The present study was designed to assess possible abuse related effects of the endogenous neuroactive steroid pregnanolone (3alpha hydroxy-5beta-pregnan-20-one) and the orally bioavailable, water-soluble neuroactive steroid pro-drug Co 8-7071 (3alpha,21-dihydroxy-3beta-trifluoromethyl 5beta-pregnan-20- one, 21-hemisuccinate). METHODS: Four rhesus monkeys were prepared with chronic intravenous (i.v.) catheters and trained to press a lever under a ten-response fixed-ratio (FR) schedule of methohexital injection (0.1 mg/kg per injection). Three rhesus monkeys were trained to discriminate intragastric infusions of pentobarbital (10 mg/kg) from saline infusions under a FR5 schedule of stimulus-shock termination. RESULTS: At least two doses of pregnanolone (0.003-0.1 mg/kg per injection) maintained injections per session above saline levels in the four monkeys tested, whereas Co 8-7071 (0.01-1.0 mg/kg per injection) maintained injections per session above saline levels in two of four monkeys at relatively low levels of injections per session. In rhesus monkeys trained to discriminate pentobarbital, i.v. pregnanolone injections (0.1 1.7 mg/kg, 5-min presession) dose-dependently reproduced the discriminative stimulus effects of pentobarbital in all monkeys tested. Intravenous administration of Co 8-7071 (1-10 mg/kg, 5-min presession) resulted in a dose dependent increase to >80% pentobarbital-appropriate responding in two of three monkeys tested. Following intragastric infusions of Co 8-7071 (1.0-30 mg/kg), > or =80% pentobarbital-appropriate responding occurred in one out of three monkeys at 10 mg/kg when administered 60 min before the session. When administered 120 min before the session, however, 10-30 mg/kg Co 8-7071 reproduced the discriminative stimulus effects of pentobarbital in each of the three monkeys tested. CONCLUSIONS: These data demonstrate barbiturate-like abuse-related effects that differed between two pregnane steroids. Whereas pregnanolone functioned as a reinforcer, suggesting that this compound has abuse potential, Co 8-7071 did not, despite having pentobarbital-like discriminative effects. PMID- 10463323 TI - Dissociating neuromodulatory effects of diazepam on episodic memory encoding and executive function. AB - RATIONALE: Diazepam and other benzodiazepines impair episodic memory encoding. Deficits in tests of executive function are also reported. In this study, we ask whether the latter effects are secondary to mnemonic impairment, or reflect specific and distinct effects of benzodiazepines on executive function. OBJECTIVES: Using positron emission tomography in healthy human volunteers, we examined similarities in the neuroanatomical correlates of the effect of diazepam on performance of executive compared to episodic memory tasks. Close similarities are proposed to reflect commonalities in the functional effects of the drug. Conversely, any evidence of task-specific regional changes in activity is proposed to reflect distinct functional effects of DZP on the two tasks. METHODS: Twelve volunteers received placebo or 10 mg diazepam in a between-subjects design. During scanning, subjects performed one of four experimental conditions, corresponding to a 2x2 factorial design, with memory encoding and executive function (on-line ordering of stimuli) as the two factors. Drug- or task-induced changes in brain activation indexed the neuroanatomical correlates of each condition. RESULTS: Averaged across all conditions, and compared to placebo, diazepam decreased activity bilaterally in prefrontal and temporal cortices. Within this network of deactivation, left dorsal prefrontal cortex activity was attenuated by diazepam during memory encoding, while left frontal opercular activity was attenuated during ordering. CONCLUSION: This neuroanatomical dissociation reflects distinct functional effects of diazepam on encoding versus ordering tasks. Therefore, the effects of diazepam on ordering tasks are not simply secondary to diazepam effects on episodic memory, but reflect real and distinct effects of the drug on executive function. PMID- 10463324 TI - Zolmitriptan-induced growth hormone release in humans: mediation by 5-HT1D receptors? AB - RATIONALE: Effective neuroendocrine probes of 5-HT1B and 5-HT1D receptor function may facilitate investigation of the role of these receptor subtypes in the pathophysiology of depression and the mode of action of antidepressant medication. OBJECTIVE: To investigate the neuroendocrine profile of the 5-HT1B/1D receptor agonist, zolmitriptan, in healthy volunteers. METHODS: Twelve subjects entered a double-blind, placebo-controlled, cross-over design study of zolmitriptan (5 mg orally). Blood samples were taken at 15-min intervals for assay of prolactin and growth hormone. A further six healthy men were recruited to an equivalent study to examine the effect of ketanserin (a 5-HT receptor antagonist with some preference for 5-HT1D over 5-HT1B receptors) on the growth hormone response to zolmitriptan. RESULTS: Zolmitriptan significantly increased plasma growth hormone but had no effect on plasma prolactin or oral temperature. The increase in growth hormone produced by zolmitriptan was significantly attenuated by ketanserin. CONCLUSIONS: We suggest that the ability of triptans such as zolmitriptan, sumatriptan and rizatriptan to increase plasma growth hormone is mediated by their common agonist activity at postsynaptic 5-HT1D receptors. PMID- 10463325 TI - Pretreatment with the putative anti-addictive drug, ibogaine, increases the potency of cocaine to elicit locomotor responding: a study with acute and chronic cocaine-treated rats. AB - RATIONALE: Results of single-dose studies suggest that the effects of pretreatment with the putative anti-addictive compound, ibogaine, on drug-induced locomotor behavior depends on the previous drug history of the animal. OBJECTIVES: To compare the effects of ibogaine pretreatment on the dose-locomotor response function for cocaine in rats treated chronically with either saline or cocaine. METHODS: Rats were chronically treated with either cocaine (15 mg/kg, IP, once daily for 5 days, followed by 2 week withdrawal) or saline. Ibogaine (40 mg/kg, IP) or vehicle was administered and 19 h later, a cocaine dose-locomotor response test was conducted (0, 5, 10, 20 and 40 mg/kg, IP). RESULTS: Chronic cocaine administration augmented the locomotor response to cocaine in chronic cocaine-treated rats, compared to acutely treated controls. Ibogaine pretreatment enhanced the locomotor effects of cocaine in both chronic and acute cocaine groups. Furthermore, due to the shape of the dose-response curve, in chronic cocaine but not in acute cocaine rats, ibogaine pretreatment enhanced the locomotor response to 5 and 10 mg/kg cocaine while decreasing the locomotor response to 40 mg/kg cocaine. CONCLUSIONS: These data demonstrate definitively that ibogaine can enhance sensitivity to the locomotor stimulant effects of cocaine, an effect which depends, in part, on the previous cocaine history of the animal. PMID- 10463326 TI - Medial prefrontal cortex is involved in the discriminative stimulus effects of nicotine in rats. AB - RATIONALE: Central nicotinic receptors have been reported to be involved in the discriminative stimulus (DS) effects of nicotine. OBJECTIVES: The purpose of the present study was to investigate the role of the medial prefrontal cortex (mPFC) and the medial habenular nucleus (mHb) in the DS effects of nicotine. METHODS: Substitution tests with nicotine administered into mPFC and mHb were conducted in rats trained to discriminate nicotine (0.5 mg/kg, SC) from saline in a two-lever, food reinforced, operant task. RESULTS: Nicotine (40 microg) administered into mPFC substituted for nicotine (0.5 mg/kg, SC), whereas nicotine administered into mHb did not. CONCLUSIONS: Together with our previous study indicating that the nucleus accumbens and the ventral tegmental area are partially involved in the DS effects of nicotine, the present study suggests that mPFC is primarily involved in the DS effects of nicotine. PMID- 10463327 TI - G-Proteins and their regulators. PMID- 10463328 TI - New insights into the in vivo function of heterotrimeric G-proteins through gene deletion studies. AB - A huge number of receptors signals through heterotrimeric G-proteins to regulate a wide variety of physiological processes. The ability to modify G-protein expression in vivo provides a powerful new tool to analyze the function of G protein-mediated signalling pathways. Genetic ablation of G-protein alpha-subunit genes is continuing to give new insights into the physiological roles of heterotrimeric G-proteins. This review highlights recent advances resulting from such genetic approaches to the study of G-protein-mediated signalling. PMID- 10463329 TI - Regulators of G-protein signalling: a novel protein family involved in timely deactivation and desensitization of signalling via heterotrimeric G proteins. AB - In a variety of signalling pathways heterotrimeric guanine-nucleotide-binding proteins (G proteins) trigger physiological responses elicited by hormones, neurotransmitters and sensory stimuli. Receptor-induced GDP/GTP exchange activates G proteins by dissociating G-protein alpha-subunits from the betagamma dimers. Both alpha-subunits and betagamma-dimers are involved in effector regulation. The deactivation of these active forms is controlled by the hydrolysis of GTP bound to alpha-subunits, allowing the inactive heterotrimer to reform. Termination of G-protein-mediated signalling in vivo is 10- to 100-fold faster than the in vitro rate of GTP hydrolysis by alpha-subunits, suggesting that in analogy to the GTPases of the Ras-superfamily, GTPase-activating proteins (GAPs) are required to achieve timely deactivation. Recently, members of a novel protein superfamily, known as "regulators of G-protein signalling" (RGS), were identified as potent GAPs for at least one subset of heterotrimeric G-protein alpha-subunits. In this review, we intend to discuss the proposed mechanism by which RGS proteins exert GAP activity for G-protein alpha-subunits as well as their specificities. The role of RGS proteins in desensitization and temporal resolution in certain signalling pathways will also be addressed. PMID- 10463330 TI - The G protein beta3 subunit splice variant Gbeta3-s causes enhanced chemotaxis of human neutrophils in response to interleukin-8. AB - A C825T polymorphism was recently described in GNB3, the gene encoding the Gbeta3 subunit of heterotrimeric G proteins. The 825T allele is associated with the expression of a shorter splice variant (Gbeta3-s) and enhanced signal transduction via pertussis toxin (PTX)-sensitive G proteins. Given the pivotal role of G protein betagamma dimers in chemotaxis, we related the genotype at the GNB3 locus as a marker for Gbeta3-s expression to chemotaxis of human neutrophils in response to stimulation with interleukin-8 (IL-8). IL-8, which activates a CXC receptor coupled to PTX-sensitive G proteins, induced at 10 nM an enhanced maximum chemotaxis of neutrophils from individuals with TC/TT genotype compared to CC genotype. Furthermore, migration of neutrophils from 825T allele carriers was 2.5-fold higher at 0.1 nM and 1 nM IL-8. At these concentrations of IL-8, no significant chemotaxis was observed in neutrophils from homozygous C825 allele carriers, indicating a genotype-dependent, different potency of IL-8 to chemoattract neutrophils. In contrast, IL-8-induced Ca2+ signals and O2- generation were independent of genotype. The role of Gbeta3-s in enhanced chemotaxis could be confirmed by determination of chemotaxis of COS-7 cells following transfection with either Gbeta3-s or "wild-type" Gbeta3. Upon stimulation of the transfected cells with the chemoattractant lysophosphatidic acid (LPA), we observed an enhanced chemotactic response of Gbeta3-s-transfected compared to Gbeta3-transfected COS-7 cells, confirming that Gbeta3-s actually causes enhanced chemotaxis. PMID- 10463331 TI - Calcium channel subtypes and exocytosis in chromaffin cells: a different view from the intact rat adrenal. AB - In the intact rat adrenal gland perfused with an oxygenated Krebs-bicarbonate solution at 37 degrees C, the electrical field stimulation of splanchnic nerves (100 V, 0.5 ms duration, 10 Hz during 10 s) produced transient catecholamine release peaks that were reproduced in subsequent stimuli, applied at 8-min intervals. Omega-Conotoxin GVIA (0.3 microM) caused only a modest inhibition of the secretory response, suggesting that the N-subtype of voltage-dependent Ca2+ channels are scarcely involved in such a response. Both omega-conotoxin MVIIC (1 microM) and furnidipine (1 microM) halved the secretion, suggesting that the L- and P/Q-types of Ca2+ channels were involved. N-type Ca2+ channels appear to be involved in the maintenance of secretion in response to sustained stimulus since omega-conotoxin GVIA (0.3 microM) reduced the catecholamine output to 28%. When secretion was elicited by acetylcholine (10 microM), furnidipine reduced the catecholamine release by 50% and omega-conotoxin MVIIC by 40%, whereas omega conotoxin GVIA did not modify the response. The K+-induced secretory responses (23.6 mM K+, 15 s) were reduced 75% by furnidipine and 45% by omega-conotoxin MVIIC, indicating that this type of stimulation preferentially recruited L-type channels. These data show that electrical stimulation recruits Ca2+ channel subtypes different from those recruited by direct depolarization of chromaffin cells. PMID- 10463332 TI - In vivo effects of the NMDA receptor agonist tetrazolyl-glycine related to the function of small-diameter primary afferents. AB - The NMDA receptor agonist tetrazolyl-glycine (TG; 100 microg kg(-1), i.v.) caused a depressor reflex in anaesthetized rats. The NMDA receptor antagonist MK-801 (300 microg kg(-1), i.v.) inhibited this depressor reflex, but not that induced by pentylenetetrazol (PTZ; 100 mg kg(-1), i.v.), indicating a selective effect of TG on NMDA receptors in vivo. Capsaicin pretreatment, which excludes the function of small-diameter primary afferent fibres, caused only a reduction of the TG induced depressor reflex, suggesting a reduction of NMDA receptors. The absence of effects of TG and PTZ on the blood pressure in pithed rats excluded any peripheral vascular actions of TG and PTZ. The depressor reflex evoked by afferent nerve stimulation was also inhibited by MK-801 (300 microg kg(-1), i.v.), but not by the tachykinin antagonist L-742694 (10 mg kg(-1), i.v.), confirming the essential role of glutamate in the neurotransmission of signals at central terminals of small-diameter afferent neurons. Plasma protein extravasation in the rat hind paw, induced by neurotransmitters released at peripheral terminals of small diameter afferent neurons by antidromic nerve stimulation, was not influenced by MK-801, indicating that glutamate is either not released or has no effect there. It is concluded that the NMDA agonist TG is a valuable tool to study the functions of primary afferents in vivo. PMID- 10463333 TI - Comparison of effects of hypotension and handling stress on the release of noradrenaline and dopamine in the locus coeruleus and medial prefrontal cortex of the rat. AB - The effects of two different types of stress (hypotension and handling) on the release of dopamine, noradrenaline and DOPAC in the locus coeruleus (LC) and medial prefrontal cortex (mPFC) was studied by means of the dual-probe microdialysis technique. One probe was implanted in the vicinity of the LC and a second probe was implanted in the mPFC. Both probes were used to record simultaneously noradrenaline, dopamine and DOPAC. Samples from the LC were collected in the presence of nomifensine, which was added to the perfusion fluid in a concentration of 50 microM. Hypotension (20 min) induced by intravenous administration of nitroprusside stimulated the release of noradrenaline in the LC and mPFC to about 190% and 150%, of control values, respectively. Hypotension also strongly stimulated the release of dopamine in the mPFC (to 320% of control) and DOPAC in the LC (to 270% of control). The effect of hypotension on extracellular dopamine, noradrenaline and DOPAC was decreased by halothane anaesthesia, and was blocked by chloral hydrate anaesthesia. Handling stress (10 min) stimulated the release of noradrenaline in the LC and mPFC to 180% and 160% of control values, respectively. Handling stimulated the release of dopamine in the mPFC to about 160% of control. The effect of hypotension or handling stress was further evaluated in animals in which the LC was lesioned by an infusion of 6 OH-dopamine. Lesioning of the noradrenergic LC neurons did not the prevent the hypotension-related stimulation of dopamine release, but shortened the time course of the effect dramatically. Lesioning of the noradrenergic neurons had no effect on the stimulatory effect of handling on the release of dopamine in the mPFC. This study shows that mesocortical dopamine neurons, in contrast to noradrenaline neurons, respond much stronger to hemodynamic stress than to an emotional stress. During certain conditions like hypotension stress, but not during handling stress, the LC activity is able to modulate the release of dopamine from mesocortical neurons. PMID- 10463334 TI - Effects of imidazoline receptor ligands on monoamine synthesis in the rat brain in vivo. AB - This study was designed to assess the effects of imidazoline drugs on putative presynaptic imidazoline receptors modulating brain monoamine synthesis in vivo. The accumulation of 3,4-dihydroxyphenylalanine (dopa) and 5-hydroxytryptophan (5 HTP) after decarboxylase inhibition was used as a measure of the rate of tyrosine and tryptophan hydroxylation in various brain regions of naive rats and after irreversible alpha2-adrenoceptor inactivation with EEDQ (1.6 mg/kg, i.p., 6 h). Clonidine (1-3 mg/kg), moxonidine (1-10 mg/kg) and rilmenidine (10 mg/kg) (mixed I1/alpha2 agonists) decreased dopa and 5-HTP synthesis in the cerebral cortex (14%-81%), hippocampus (27%-84%) and/or striatum (29%-56%), but these inhibitory effects were abolished in N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) treated rats. Similarly, the stimulatory effect of efaroxan (mixed I1/alpha2 antagonist; 10 mg/kg) on dopa synthesis in the cortex (77%) and hippocampus (57%) was abolished by EEDQ. The selective I1-ligand 2-endo-amino-3-exoisopropylbicyclo heptane (AGN-192403; 5-10 mg/kg) did not modify dopa or 5-HTP synthesis in any brain region in naive or EEDQ-treated rats. Idazoxan (mixed I2/alpha2 antagonist; 20 mg/kg) increased dopa synthesis in the cortex (111%) and hippocampus (87%), but the stimulatory effects were abolished by EEDQ. Moreover, idazoxan and efaroxan decreased 5-HTP synthesis in the cortex (12%-34%) and hippocampus (30% 34%) in a manner sensitive to blockade by the 5-HT1A receptor antagonist WAY 100135. The selective I2-igands 2-(2-benzofuranyl)-2-imidazoline (2-BFI; 20 mg/kg) and 2-styryl-2-imidazoline (LSL 61122; 10 mg/kg) did not alter the synthesis of dopa or 5-HTP in the cortex or hippocampus. In striatum, 2-BFI (1-20 mg/kg) dose-dependently decreased dopa synthesis (ED50: 5.9 mg/kg), reduced dopamine levels (6%-36%) and increased those of its metabolites DOPAC (15%-95%) and HVA (24%-74%). The inhibitory effect of 2-BFI on dopa/dopamine synthesis in striatum remained unchanged after alkylation of imidazoline receptors with isothiocyanatobenzyl imidazoline (IBI; 60 mg/kg, 6 h) or blockade of these receptors with 2-(2-ethyl 2,3-dihydro-2-benzofuranyl)-2-imidazole (KU-14R; 7-20 mg/kg). Therefore, most imidazoline drugs modulated the synthesis of brain monoamines through interaction with alpha2-adrenoceptors or 5-HT1A receptors. The results do not provide functional evidence for the existence of presynaptic imidazoline receptors regulating the synthesis of monoamines in the rat brain. PMID- 10463335 TI - Interleukin-1beta-induced nitric oxide synthesis in aortic rings from normal and hyperinsulinaemic rats: effect of physical exercise. AB - Insulin has been suggested to prevent the induction of nitric oxide synthase (NOS) in vitro in arterial smooth muscle, but whether such a mechanism is operative in vivo is not known. Therefore, we evaluated the sensitivity of smooth muscle NOS to induction by interleukin-1beta (IL-1beta) in aortic rings of lean and obese Zucker rats, a model of experimental hyperinsulinaemia. In order to modulate the insulin and glucose balance of the rats, a 22-week-long treadmill exercise was included in the study. The training attenuated weight gain and reduced blood glucose in the obese and lean rats, whereas the abnormally high plasma insulin of the obese rats remained unaffected. A 6-h incubation of aortic rings with IL-1beta (10 ng/ml) increased cyclic GMP in smooth muscle by approximately threefold in all groups, and this effect was prevented by methylene blue. The contractile sensitivity of endothelium-denuded aortic rings to phenylephrine was reduced by incubation with IL-1beta (1 ng/ml and 10 ng/ml) in the exercised obese and lean rats, whereas no significant change was observed in the sedentary groups. The aortic maximal contractile force induced by phenylephrine was reduced in sedentary and exercised obese rats by incubation with IL-1beta, while no change was detected in the lean rats. The aortic relaxation to exogenous L-arginine was augmented by IL-1beta in all groups, while the relaxation sensitivity to L-arginine after induction by IL-1beta was enhanced by exercise in the obese but not in the lean rats. Finally, the relaxation to nitroprusside was not significantly affected by IL-1beta in any of the study groups. In conclusion, since maximal contractile force generation to phenylephrine was reduced by IL-1beta in the obese but not in the lean rats, the sensitivity of NOS to induction by IL-1beta was higher in arterial smooth muscle of the obese than the lean Zucker rats. Thus, this model of hyperinsulinaemia was not associated with reduced sensitivity of smooth muscle NOS to induction by IL 1beta. Regular exercise did not change plasma insulin concentrations, but it enhanced the action of insulin in both strains as reflected by reduced blood glucose, and increased the sensitivity of smooth muscle NOS to induction by IL 1beta. PMID- 10463337 TI - Differential mechanisms of urethral smooth muscle relaxation by several NO donors and nitric oxide. AB - We have examined the mechanisms of action of a broad spectrum of nitric oxide (NO) donors, including several S-nitrosothiols, sodium nitroprusside (SNP) and nitroglycerine (GTN), in relation to their relaxant activity of urethral smooth muscle. For all the compounds examined, NO release (in solution and in the presence of urethral tissue), relaxation responses, elevations in cGMP levels and the effect of thiol modulators were evaluated and compared with the effect of NO itself. Whilst all NO donors, except GTN, released NO in solution due to photolysis or chemical catalysis, this release was not correlated with their relaxant activity in sheep urethral preparations, which were furthermore not affected by the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1 oxyl 3-oxide (cPTIO; 0.3 mM). A substantial NO-generating activity was found for S-nitroso-L-cysteine (CysNO) and S-nitroso-N-acetyl-D,L-penicillamine (SNAP) in the presence of urethral cytosolic fractions, suggesting metabolic activation to NO in the cytosol of the target tissue. In contrast, NO generation from S-nitroso N-acetyl-L-cysteine (N-ac-CysNO), S-nitrosoglutathione (GSNO) and SNP were reduced by the presence of urethral homogenate and/or subcellular fractions, suggesting direct NO transfer to tissue constituents. NO donors and NO gas induced dissimilar degrees of cGMP accumulation in urethral tissue, while they were essentially equipotent as urethral relaxants. Furthermore, 1H-[1,2,4] oxadiazole-[4,3-a]-quinoxalin-1-one (ODQ; 10 microM) inhibited both relaxation and cGMP accumulations, but with different potency for the different compounds. Oxidation of sarcolemmal thiol groups with 5-5'-dithio-bis[2-nitrobenzoic acid] (DTNB; 0.5 mM) enhanced relaxations to GSNO, an effect that was reversed by dithiotreitol (DTT; 1 mM), suggesting a direct effect through nitrosylation/oxidation reactions at the cell membrane, while relaxations to NO and to all the other compounds were not affected by these treatments. Finally, photodegradation of SNP induced the formation of a stable intermediate that still evoked NO-cGMP-mediated relaxations. This indicates that the assumption that SNP is fully depleted of NO by exposure to light should be revised. It can be concluded that important differences exist in the mechanisms by which distinct NO donors relax urethral smooth muscle and they cannot be regarded simply as NO releasing prodrugs. PMID- 10463336 TI - Evidence for a contribution of store-operated Ca2+ channels to NO-mediated endothelium-dependent relaxation of guinea-pig aorta in response to a Ca2+ ionophore, A23187. AB - A23187 (6S-[6alpha,8beta,9beta,11alpha]-5-(methylamino) -2-[[3,9,11-trimethyl-8 [1-methyl-2-oxo-2-(1H-pyrrol-2-yl)ethyl]-1,7- dioxaspiro[5.5]undec-2-yl]methyl]-4 benzoxazolecarboxylic acid, calcimycin), an antibiotic Ca2+ ionophore, produces an endothelium-dependent vascular relaxation. In the present study, pharmacological features were functionally characterized of endothelium-dependent relaxant response of guinea-pig aorta to A23187, especially focusing on the possible Ca2+ source and Ca2+ mobilization mechanisms in endothelial cells responsible for the vasorelaxant response to the Ca2+ ionophore. A23187-induced endothelium-dependent relaxation was suppressed profoundly by N(G)-nitro-L arginine (L-NNA; 3 x 10(-4) M) or calmidazolium (3 x 10(-5) M), suggesting that nitric oxide (NO) produced by the enhanced activation of Ca2+/calmodulin dependent endothelial NO synthase (eNOS) is largely responsible for the relaxant response of this artery to A23187. In the Ca2+-free solution without EGTA, NO mediated endothelium-dependent relaxation induced by A23187 was almost abolished, which suggests that Ca2+ entry from extracellular space into endothelial cells plays the key role in the A23187-induced functional vasorelaxation. On the other hand, SK&F96365 (1-[beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H imidazole; 5 x 10(-5) M) and Ni2+ (3 x 10(-4) M), both of which inhibit capacitative Ca2+ influx through store-operated Ca2+ channels (SOCCs), attenuated significantly NO-mediated endothelium-dependent relaxation by A23187. Furthermore, A23187-induced endothelium-dependent relaxation was suppressed more strongly than endothelium-independent relaxation induced by SIN-1 (3-morpholino sydnonimine), an NO donor, when aortic preparation was preconstricted with high KCl instead of agonistic stimulation (prostaglandin F2alpha). These findings suggest that NO-mediated endothelium-dependent relaxant response of guinea-pig aorta to A23187 is preceded by the increase in endothelial cytosolic free Ca2+ concentration ([Ca2+]cyt) due to the enhanced Ca2+ influx from extracellular space. In the enhanced Ca2+ entry leading to the stimulation of eNOS and NO mediated functional relaxant response of guinea-pig aorta to A23187, activation of SOCCs but not the Ca2+ entry through plasma membrane Ca2+-specific routes made by A23187 seems to play the predominant role. It is most likely that A23187 acts primarily at the Ca2+ store sites in endothelial cells, which subsequently depletes stored Ca2+ to activate SOCCs via unidentified mechanisms. PMID- 10463338 TI - Increase in histidine decarboxylase activity in tissues of mice bearing Colon-26 tumor cells. AB - The changes in histidine decarboxylase (HDC) activity, histamine and tele methylhistamine contents were examined in tissues of mice after the inoculation of Colon-26 tumor cells subcutaneously into the lower back. The HDC activity in the spleen of mice increased significantly 14 days after the inoculation of Colon 26 and the increase in HDC activity continued for up to 28 days. However, the histamine content in the spleen of tumor-bearing mice was not changed significantly during the observation period. In the following experiments, two subclones of the Colon-26 cell line, cachexia-inducing clone-20 and non cachexia inducing clone-5, were used and the induction of HDC activity in mice was examined in four tissues, spleen, lung, liver and kidney. Both clone-20 and clone 5 induced the increase in HDC activity to the same extent in the spleen and lung, but not in the liver and kidney. As observed using the Colon-26 original cell line, the histamine contents in the four tissues of tumor-bearing mice were not different from those in the control mice. In contrast, the levels of tele methylhistamine, one of the major catabolites of histamine, in the tumor-bearing mice increased significantly compared with the control mice in all four tissues examined. There was a correlation between the increase in tele-methylhistamine level and the increase in HDC activity in the tissues. A histological study indicated that the tissue mast cells were not increased in spleen and lung of tumor-bearing mice. These findings indicated that the increase in HDC activity in the spleen and lung occurred in parallel with the growth of inoculated tumor cells in mice and suggested that the cells other than mast cells may be involved in the increase in HDC activity. The tumor-bearing state produced histamine with a high turnover rate in the mouse tissues, especially in the spleen and lung. PMID- 10463339 TI - Modification of low density lipoprotein potentiates its inhibitory effect on catecholamine secretion in cultured bovine adrenal medullary cells. AB - Low density lipoprotein (LDL) and lipoprotein(a) suppress catecholamine secretion in cultured adrenal medullary cells. Modification of LDL by oxidation or acetylation potentiates various atherogenic actions of LDL. In the present study, we investigated whether the modification of LDL influences catecholamine secretion in cultured bovine adrenal medullary cells. The exposure of LDL to CuSO4 caused a time-dependent oxidation of LDL. Maximal oxidation of LDL was observed after exposure to CuSO4 for 24 h. Native LDL inhibited catecholamine secretion induced by carbachol to 68.5% of control. Oxidized LDL caused further inhibition of carbachol-evoked secretion to 37.6% of control. Acetylated LDL inhibited it to 41.0% of control. There was a good correlation between the extent of LDL oxidation and the inhibition of catecholamine secretion. These results suggest that oxidation or acetylation of LDL augments its inhibitory effect on the secretion of catecholamines. Since catecholamines are a risk factor of atherosclerosis, the inhibitory effect by such modified LDL may be a mechanism inhibiting atherosclerotic progression. PMID- 10463340 TI - Sonic hedgehog enhances somite cell viability and formation of primary slow muscle fibers in avian segmented mesoderm. AB - Primary skeletal muscle fibers first form in the segmented portions of paraxial mesoderm called somites. Although the neural tube and notochord are recognized as crucial in patterning myogenic cell lineages during avian and mammalian somitic myogenesis, the source, identities, and actions of the signals governing this process remain controversial. It has been shown that signals emanating from the ventral neural tube and/or notochord alone or Shh alone serve to activate MyoD expression in somites. However, beyond a role in initiating MyoD expression, little is known about the effects of Shh on primary muscle fiber formation in somites of higher vertebrates. The studies reported here investigate how the ventral neural tube promotes myogenesis and compare the effects of the ventral neural tube with those of purified Shh protein on fiber formation in somites. We show that purified Shh protein mimics actions of the ventral neural tube on somites including initiation of muscle fiber formation, enhancement of numbers of primary muscle fibers, and particularly, the formation of primary fibers that express slow myosin. There is a marked increase in slow myosin expression in fibers in response to Shh as somites mature. The effects of ventral neural tube on fiber formation can be blocked by disrupting the Shh signaling pathway by increasing the activity of somitic cyclic AMP-dependent protein kinase A. Furthermore, it was demonstrated that apoptosis is a dominant fate of somite cells, but not somitic muscle fibers, when cultured in the absence of the neural tube, and that application of Shh protein to somites reduced apoptosis. The block to apoptosis by Shh is a manifestation of the maturity of the somite with a progressive increase in the block as somites are displaced rostrally from somite III forward. We conclude that purified Shh protein in mimicking the effects of the ventral neural tube on segmented mesoderm can exert pleiotropic effects during primary myogenesis, including: control of the proliferative expansion of myogenic progenitor cells, antagonism of cell death pathways within the precursors to muscle fibers, and during the crucial process of primary myogenesis, can exert an effect on diversification of muscle fiber types. PMID- 10463341 TI - Localization of micro- and intermediate filaments in non-pregnant uterus and placenta of the mink suggests involvement of maternal endothelial cells and periendothelial cells in blood flow regulation. AB - Maternal stem arteries and arterioles of the endotheliochorial mink placenta have been shown to lack smooth muscle cells, suggesting a muscle-free attenuation of the maternal arterial pulse wave of the placenta. Since the endotheliochorial type of placenta by definition does not contain any maternal supportive tissue (e.g. connective tissue), except for the specialized interstitial layer, the aim of this study was to reveal cytoskeletal components able to compensate for the lack of conventional regulatory mechanisms of maternal placental blood flow. The study was undertaken on buffered formalin fixed tissues from 19 minks by immunohistochemistry and transmission electron microscopy to localize three major cytoskeletal filaments (desmin, vimentin and alpha-smooth muscle actin (alpha-sm actin)) in non-pregnant uteri and placenta. The contractile alpha-sm-actin was immunodetected in the maternal subepithelial and periglandular connective tissue cells of the cyclic endometrium and during early gestation. During the transition from early- to mid gestation, maternal periendothelial cells appeared and showed alpha-sm-actin immuno-positivity; however, in late gestation, this activity could not be detected because the periendothelial cells had disappeared. Fetal endothelial cells displayed intense alpha-sm-actin immunoreactivity, which was in contrast to the alpha-sm-actin negative maternal endothelial cells. Allantochorionic mesenchymal cells also exhibited intense alpha-sm-actin immunostaining. Vimentin was immunohistochemically expressed in endothelial cells (maternal as well as fetal), maternal periendothelial cells, allantochorionic mesenchymal cells, and maternal connective tissue cells from early gestation. Desmin was not immunohistochemically detectable in cyclic endometrium and placental tissues. Transmission electron microscopy revealed the periendothelial cells to be enclosed by a thin interstitial layer. Additionally, the maternal endothelial cells displayed actin myofilament-like structures anchored basally. From our data we conclude that maternal periendothelial cells, immunoreactive for contractile actin, and maternal endothelial cells, possessing actin myofilament like ultrastructures, act as supportive systems in the maternal vessel walls, probably influencing the regulation of the maternal blood flow. PMID- 10463342 TI - Nitric oxide synthase-containing projections to the ventrobasal thalamus in the rat. AB - Microiontophoretic studies of thalamic neurons suggests that nitric oxide (NO) plays an important role in mediating somatosensory transmission. The thalamus contains few nitric oxide synthase (NOS)-immunoreactive neurons; thus, the major source of thalamic NO is presumably from NOS-positive axons of extrathalamic origin. The cells of origin of these putative NOS-containing pathways to the ventrobasal thalamus were investigated in rats by combining retrograde tracing with immunocytochemistry for NOS. The location and morphology of double-labeled neurons was compared with that of single-labeled neurons. The most significant sources of NOS-containing afferents to the thalamus were found to be the pedunculopontine (PPN) and laterodorsal tegmental (LDT) nuclei. NOS immunoreactive neurons in these cholinergic nuclei project bilaterally to the thalamus, most strongly ipsilaterally. The thalamus appears to be a major target of PPN, since even selective thalamic injections result in retrograde labeling of at least one third of its NOS-immunoreactive neurons. A significant number of NOS negative neurons in both the PPN and LDT also project to the thalamus. Minor sources of NOS-containing thalamic afferents include the lateral hypothalamus, the dorsal, median and pontine raphe nuclei, the parabrachial nuclei, and the pontomedullary reticular formation. In all these structures, NOS-negative thalamopetal neurons greatly outnumber the NOS-positive ones. Ascending sensory pathways to the thalamus, including those from the sensory trigeminal nuclei, the dorsal column nuclei, and the spinal cord, as well as the auditory and vestibular centers, arise exclusively from NOS-negative neurons. The major NOS-positive projections are implicated in affective and alerting systems, supporting that NO may act to modulate attentiveness in thalamic relay nuclei. PMID- 10463343 TI - In situ detection of gelatinolytic activity in developing craniofacial tissues. AB - Frozen tissue sections of developing and adult rat heads were incubated on a film coated with a gelatin-containing colloidal silver emulsion in order to detect gelatinolytic activity present in the different tissues. The method, termed film in situ zymography, is based on the ability of the thiol group of the propeptide released from the degraded gelatin to induce a structural change in the colloidal silver and thereby a visible change in color. The frozen tissue sections mounted on the coated film were incubated at 37 degrees C overnight. Gelatinolytic activity was detected as a color change from yellow to red. The activity of gelatinase was completely blocked by phenanthroline, which inhibits matrix metalloproteinases. Gelatinolytic activity was widely present in the oral epithelium, tooth buds, tongue, Meckel's cartilage, salivary glands, and other tissues. The intensity of the gelatinolytic activity varied among the different tissue types. The present study demonstrated gelatinolytic activity in both developing and adult craniofacial tissues. These results suggest that gelatinolytic activity plays an important role in normal turn-over in several tissues. Whereas some of the activity also in the developing rats may be related to this turn-over process, some of it is probably directly associated with developmental changes. PMID- 10463344 TI - Proliferation of actively migrating ameboid microglia in the developing quail retina. AB - Sheets containing the inner limiting membrane covered by a carpet of Muller cell endfeet were used to show that ameboid microglial cells migrating tangentially in the vitreal part of the developing retina of quail embryos underwent mitosis. Double labeling with anti-beta-tubulin/QH1 or Hoechst 33342/QH1 revealed that some migroglial cells with morphological features typical of active migration were in early prophase. By anaphase and early telophase, microglial cells had retracted their lamellipodia and were ovoid in shape. Later in telophase, but well before completion of cytokinesis, both daughter cells again emitted lamellipodia, thus regaining the typical morphology of migrating cells. We concluded that ameboid microglial cells go through cycles in which migration and mitosis alternate, and that both mechanisms contribute to the spread of microglia throughout the developing retina. The mitotic spindle of dividing microglial cells showed different orientations, which probably influenced the course of subsequent migration. The expression of the proliferating cell nuclear antigen in the nucleus of most tangentially migrating ameboid microglial cells at E9-E10 confirmed their proliferative capability. However, the rate of proliferation of these cells decreased during embryonic development, and was nearly zero at E14. PMID- 10463345 TI - Synaptology and ultrastructural characteristics of laryngeal cricothyroid and posterior cricoarytenoid motoneurons in the nucleus ambiguus of the rat. AB - The laryngeal motoneurons innervating the cricothyroid muscle (CT) are located in the semicompact formation just ventral to the rostral part of the compact formation of the nucleus ambiguus. The motoneurons innervating the posterior cricoarytenoid muscle (PCA) are located in the loose formation. We retrogradely labeled the CT and the PCA motoneurons using cholera toxin subunit B-conjugated horseradish peroxidase, and determined the ultrastructure and synaptic organization of these neurons. The CT and the PCA motoneurons had the appearance of alpha-motoneurons, i.e., large, oval or polygonal cells containing well developed organelles and a prominent spherical nucleus. Two kinds of neurons were recognized among the PCA motoneurons. The one (PCA-A) was significantly smaller than the other (PCA-B). The average number of axosomatic terminals in a section was significantly largest in the PCA-B (56.6), smaller in the PCA-A (36.0), and smallest in the CT (32.3) neurons. Most of the axosomatic terminals (64.7%) contained pleomorphic vesicles and made symmetric synaptic contacts (Gray's type II) with the PCA-A neurons, while more than 60% contained round vesicles with asymmetric synaptic contacts (Gray's type I) in the CT (69.5%) and the PCA-B (60.6%) neurons. A few terminals associated with subsurface cisterns were present on all laryngeal motoneurons. These results indicated that the CT motoneurons may receive mostly excitatory terminals, whereas the PCA muscle may be regulated by neurons having many inhibitory terminals, and neurons having many excitatory terminals. PMID- 10463346 TI - Intima-like smooth muscle cells: developmental link between endothelium and media? AB - The presence of non-contractile smooth muscle cells within the arterial wall raises questions as to their origin and function. These cells abound within the aortae of murine and porcine neonates, but are also present within the intimal and medial layers of adult arteries. They are largely devoid of smooth muscle associated proteins and manifest an epithelioid form. Their morphological resemblance to endothelial cells prompted us to explore this potential relationship and to investigate their angiogenic properties in three-dimensional collagen gels. Using well-characterized smooth muscle cell lines, displaying either the intima-like (epithelioid) or media-like (spindle-shaped) morphology, we were able to show that intima-like cells share several features in common with endothelial ones and can transform into a media-like phenotype, whereby they irreversibly lose their characteristic pattern of protein expression. Intima like, but not media-like, vascular smooth muscle cells are capable of forming capillary tubes, and, in co-cultures, can induce media-like ones to participate in this process. Such capillaries consist of a randomly-organized, mixed population of endothelial cells with intima-like or media-like smooth muscle ones. The functional significance of this diversity in smooth muscle cell type is not well understood, but phenotypic plasticity could conceivably figure as an important adaptive response to changes in the local environment. PMID- 10463347 TI - Structure and vascularization of the cruciate ligaments of the human knee joint. AB - The structure and vascularization of the human anterior and posterior cruciate ligament were investigated by light microscopy, transmission electron microscopy,, injection techniques and by immunohistochemistry. The major part of the anterior and posterior cruciate ligament is composed of bundles of type I collagen. Type III collagen-positive fibrils separate the bundles. The major cell type is the elongated fibroblast, lying solitarily between the parallel collagen fibrils. The histologic structure of the cruciate ligaments is not homogeneous. In both ligaments there is a zone where the tissue resembles fibrocartilage. In the anterior cruciate ligament the fibrocartilaginous zone is located 5-10 mm proximal of the tibial ligament insertion in the anterior portion of the ligament. In the posterior cruciate ligament the fibrocartilage is located in the central part of the middle third. Within those zones the cells are arranged in columns and the cell shape is round to ovoid. Transmission electron microscopy reveals typical features of chondrocytes. The chondrocytes are surrounded by a felt-like pericellular matrix, a high content of cellular organelles and short processes on the cell surface. The pericellular collagen is positive for type II collagen. The major blood supply of the cruciate ligaments arises from the middle geniculate artery. The distal part of both cruciate ligaments is vascularized by branches of the lateral and medial inferior geniculate artery. Both ligaments are surrounded by a synovial fold where the terminal branches of the middle and inferior arteries form a periligamentous network. From the synovial sheath blood vessels penetrate the ligament in a horizontal direction and anastomose with a longitudinally orientated intraligamentous vascular network. The density of blood vessels within the ligaments is not homogeneous. In the anterior cruciate ligament an avascular zone is located within the fibrocartilage of the anterior part where the ligament faces the anterior rim of the intercondylar fossa. The fibrocartilaginous zone of the middle third of the posterior cruciate ligament is also avascular. According to Pauwel's theory of the "causal histogenesis" (1960) the stimulus for the development of fibrocartilage within dense connective tissue is shearing and compressive stress. In the anterior cruciate ligament this biomechanical situation may occur when the ligament impinges on the anterior rim of the intercondylar fossa when the knee is fully extended. Compressive and shearing stress in the center of the middle third of the posterior cruciate ligament may result from twisting of the fiber bundles. PMID- 10463348 TI - Lack of correlation between placenta and offspring size in mouse interspecific crosses. AB - The placenta plays a pivotal role in fetal growth control and is considered a major site of genetic conflict between maternal and paternal genomes within the conceptus and, in addition, the genome of the mother. Accordingly, placental development is a strictly controlled process, and both placental and fetal weights do not vary much in intraspecific crosses of laboratory mice (Mus musculus). In mouse interspecific crosses and backcrosses [(M. musculus x M. spretus) x M. musculus], tremendous variation of placental, but not of fetal weight was observed. We have studied trophoblast cell type distribution and differentiation, and their effect on the associated placentas and fetuses in such backcrosses. Differentiation of spongious trophoblast, but not size of materno fetal interface, correlated with fetal weight. Giant fetuses were observed only if less than one third of the spongiotrophoblast was formed by glycogen cells. Thus, placental efficiency was inversely related to the amount of glycogen cells. This influence of a trophoblast-derived cell type on fetal growth was not anticipated. We conclude that: (1) glycogen cells are able to negatively modulate fetal growth by an as yet unidentified mechanism; (2) correlation between fetal and placental weights is weak or absent in interspecific hybrids; (3) impaired control over placental and embryonic development in hybrids may contribute to post-mating isolation of species. PMID- 10463349 TI - Antimigraine drugs. AB - Migraine is a paroxysmal disorder characterized by attacks of headache, nausea, vomiting, photophobia, phonophobia, and malaise. This review summarizes new treatment options for therapy of the acute attack. Mild or moderate migraine attacks are treated with antiemetics followed by analgesics such as aspirin, paracetamol, nonsteroidal anti-inflammatory drugs, or antiemetics combined with ergotamine or dihydroergotamine. Sumatriptan, a specific serotonin (5-HT)1B/D agonist is used when attacks do not respond to ergotamine, or when intolerable side effects occur. The new migraine drugs zolmitriptan, naratriptan, rizatriptan, and eletriptan differ slightly in their pharmacological profiles, which translates into minor differences in efficacy, headache recurrence, and side effects. New drugs in migraine prophylaxis include cyclandelate, valproic acid and magnesium. PMID- 10463350 TI - Electrophysiological diagnosis of motor neuron disease and pure motor neuropathy. AB - Motor neuron disease (MND) is a group of disorders in which there is degeneration of upper and lower motor neurons to a variable degree. Amyotrophic lateral sclerosis is the most frequent form of the disease, presenting with both upper and lower motor neuron involvement. Frequently, especially in the early stages of the disease, only lower motor neuron signs are present. In these conditions, some pure motor neuropathies may resemble MND. The diagnosis is of importance because some of these motor neuropathies are "dysimmune" disorders and may respond to immune therapies. In such diseases the multifocal motor neuropathy with conduction block appears to be the more frequent. In MND and pure motor neuropathies, the electrophysiological examination is the most decisive test. In MND, it is of diagnostic importance. In addition, it is useful in the assessment of disease severity and progression, in the evaluation of therapeutic trials and in the understanding of etiopathogenesis of the disease. In pure motor neuropathies, the presence of conduction block leads to immune treatment with good response in more than 50% of the cases. PMID- 10463351 TI - Muscle and skin biopsies are a sensitive diagnostic tool in the diagnosis of CADASIL. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a microangiopathic syndrome. Although the defective gene has been identified, genetic analysis may be effort some due to its large size and various mutations. Providing a reliable diagnostic marker would therefore be helpful. Electron microscopy has revealed characteristic electron-dense granular deposits in the basal lamina of vessels of patients with CADASIL. We investigated the sensitivity of skin and muscle biopsies for diagnosing CADASIL. We examined 30 family members of three unrelated German families affected by CADASIL. In 14 of the 21 affected individuals we performed skin and muscle biopsies; two patients were clinically asymptomatic. Under electron microscopy all muscle and skin biopsy specimens showed patches of granular and electron-dense material in the basal layer of both arterioles and capillaries. These findings confirm that general microangiopathy is a typical feature of this syndrome and is present in the early phase of the disease with or without clinical manifestation. Thus, as electron microscopy of skin biopsy specimens can establish the diagnosis of CADASIL with high certainty, it may be considered the method of first choice. PMID- 10463352 TI - Nonenzymatic antioxidants of blood in multiple sclerosis. AB - Free radical action has been suggested as a causal factor in multiple sclerosis. We investigated the plasma level of lipid peroxides expressed in terms of malone dialdehyde and changes in blood nonenzymatic antioxidants (glutathione, alpha tocopherol, retinol, plasma sulfhydryl groups, and uric acid) in multiple sclerosis patients with exacerbation or in remission, including a group treated with beta-interferon. The malone dialdehyde level was increased by 38% (n.s.) during exacerbations. The blood concentration of oxidized glutathione was likewise elevated (P<0.05), while the ratio of plasma alpha-tocopherol to cholesterol plus triglyceride was decreased (P<0.001). These changes suggest increased free radical production and consumption of the scavenger molecules during the active phase of the disease. Blood reduced glutathione level was increased (P<0.01) during exacerbation and remission as well. The rise in this thiol is likely to be a compensatory mechanism defending the cells from further oxidant injuries. Beta-interferon increased plasma alpha-tocopherol levels (P<0.001) but not the lipid corrected alpha-tocopherol value. Other parameters were not influenced by the drug. PMID- 10463353 TI - Primitive reflexes in a case-control study of patients with advanced human immunodeficiency virus type 1. AB - This study estimated the frequency of nine primitive reflexes (PR) and assessed their possible clinical value in a group of patients with acquired immunodeficiency syndrome. We studied 78 patients with human inmunodeficiency type 1 (HIV-1) infection in WHO clinical stage 3 or 4 and 81 matched seronegative controls. All participants were examined using a standardized neurological examination and the Mini-Mental State Examination. Cognitive impairment and PR was found in 36% of patients but in none of the controls (P<0.0001; logistic regression odds ratio: 14.7). Overall, PR were 2-36 times more frequent in patients with HIV-1 infection. This association was stronger for the glabellar, snout, Rossolimo, and digital signs. At least two PR were observed in 92% of patients vs. 8% of controls (P<0.0001; 95% confidence interval: 68%-100%; logistic regression odds ratio: 10.8). These data support the association of PR with cognitive decline in patients with advanced HIV-1 infection without overt neurological disease. Larger follow-up studies with multivariate techniques are needed to identify which PRs are useful as indicators of HIV-1-associated cognitive/motor complex and minor neurocognitive disorders. PMID- 10463354 TI - Median nerve mononeuropathy in spondylotic cervical myelopathy: double crush syndrome? AB - We studied the association between spondylotic cervical myelopathy (SCM) and median nerve mononeuropathy (MNM) and examined the validity of the double-crush hypothesis. Sixty consecutive patients with clinically overt spondylotic cervical myelopathy were examined by means of nerve conduction studies, electromyography, and median nerve somatosensory evoked potentials; the frequency of the electrophysiological signs of focal MNM at the wrist was compared with that of a control group comprising 100 sex- and age-matched patients. Electrophysiological signs of MNM were found in 20 myelopathic patients (33%) in comparison with an 11% prevalence in the control group (P<0.05). The signs of motor anterior horn cell lesion at the C8-Th1 level and concomitant motor axonal MNM ipsilaterally were found in three hands, while the signs of sensory axonal loss at C6-7 segments due to ganglionic or postganglionic sensory lesion outside the wrist and concomitant sensory axonal MNM were present in one hand. While demonstrating a statistically significant association between SCM and MNM, we found no evidence of an etiological relationship between these two conditions. Electrophysiological signs of MNM fail anatomical (segmental level and side) and pathophysiological (axonal type of lesion) requirements of the double-crush hypothesis in most of patients with concomitant SCM and MNM. PMID- 10463355 TI - Varying occurrence of vocal cord paralysis in a family with autosomal dominant hereditary motor and sensory neuropathy. AB - A white British family with the axonal form of hereditary motor and sensory neuropathy (HMSN, type II) contained one member who developed a recurrent laryngeal nerve palsy at the age of 41 years, in addition to 4 years of symptomatic polyneuropathy and an abducens nerve palsy. Neither of the other family members (the mother and sister) with electrophysiologically confirmed polyneuropathy had any neuropathic symptoms in the limbs or laryngeal or respiratory muscle involvement. An autosomal dominant pattern of inheritance is likely. This is a second report of this rare form of HMSN (type IIC) in which there is associated laryngeal or respiratory muscle weakness. This family differs from the two previously reported pedigrees in which laryngeal or diaphragm weakness had commenced within the first two decades. The discovery of asymptomatic family members attests to the diagnostic value of clinical and electrophysiological study of first-degree relatives when laryngeal or bulbar symptoms develop in the context of chronic axonal polyneuropathy. HMSN type IIC should be distinguished from the more common forms of HMSN - type IIA, in which axonal polyneuropathy is restricted to the limbs, and type IIB, which is of early onset and associated with foot ulceration. PMID- 10463356 TI - Troyer syndrome: a combination of central brain abnormality and motor neuron disease? AB - Hereditary spastic paraplegia is a group of clinically and genetically heterogeneous disorders consisting of pure and complicated forms. A variant with the additional features of severe atrophy of the small hand muscles, dysarthria, mental retardation, and short stature has been termed Troyer syndrome (MIM#275900) after the name of Old Order Amish families suffering from these symptoms. We report here an Austrian family with two individuals who exhibit all the features of Troyer syndrome, and provide additional data on this disorder. Electrophysiological studies showed chronic denervation and reduced motor nerve conduction velocities but normal sensory potentials. Muscle biopsy revealed a neurogenic pattern while the sural nerve was normal on histological examination. Brain abnormalities on magnetic resonance imaging consisted of a thin corpus callosum with a poorly developed cingulate gyrus and mild periventricular signal hyperintensities. These findings characterize the Troyer syndrome as a disorder of the first and second motor neuron with additional damage in the brain. The morphological features observed in this family may contribute to the grouping and subsequent understanding of complicated forms of hereditary spastic paraplegia, together with similar observations in other, more recently reported families. PMID- 10463357 TI - Problems in designing and recruiting to therapeutic trials in primary progressive multiple sclerosis. AB - Patients with primary progressive multiple sclerosis have atypical clinical and magnetic resonance imaging (MRI) characteristics which present unique problems in designing and recruiting to therapeutic trials. The first randomised controlled therapeutic trial specifically for primary progressive multiple sclerosis is now underway. Although only an exploratory phase II study, it has provided further insight into difficulties in diagnosis, classification and choice of clinical and MRI outcome measures. Patients with primary progressive multiple sclerosis have a wide differential diagnosis and do not readily conform to the Poser criteria. They may therefore present diagnostic uncertainty, particularly as their classification often relies on a retrospective history. This was highlighted during the recruitment to this study of interferon-beta1a. Of the 138 patients referred with a definite diagnosis of primary progressive multiple sclerosis only 50 were enrolled in the study. Of the 88 patients not included, 50% either did not have primary progressive multiple sclerosis, or the diagnosis was not secure. Outcome measures pose particular problems. Clinically the focus must be on progression, and the measure should be both responsive and reliable. In relation to MRI, the currently recommended measures for therapeutic trials in relapsing/remitting and secondary progressive multiple sclerosis show little change in primary progressive multiple sclerosis, and therefore more pathologically specific MRI measures are required. Strict clinical guidelines and further developments in clinical and MRI measures are required to facilitate future therapeutic trials in primary progressive multiple sclerosis. PMID- 10463359 TI - Apolipoprotein E and presenilin-1 genotypes in Huntington's disease. AB - Huntington's disease (HD) is an autosomal dominant degenerative disease of the central nervous system manifested by involuntary movements (chorea), psychiatric manifestations, and cognitive impairment with a variable age at onset. This variability is mainly attributed to genetic factors. The so-called aging genes [e.g., those for apolipoprotein E (APOE) and presenilin-1 (PS-1) have been implicated in determining the age at onset of Alzheimer's disease, a disease sharing common clinical features with HD. In 60 unrelated patients suffering from HD (mean age at onset 40.1 years, range 20-65) we determined number of CAG repeats and the distribution of the APOE alleles (epsilon2, epsilon3, epsilon4) and PS-1 alleles. The results showed that: (a) The age at onset was higher in the group of patients with the epsilon4 allele (51.6 vs. 38.0 P<0.002), (b) The correlation between the age at onset and the number of CAG repeats was strong in patients with the epsilon3/epsilon3 genotype while it was not detected in patients with epsilon3/epsilon4 genotype. (c) No correlation was found between age at onset and PS-1 alleles. In conclusion, APOE seems to be a significant factor influencing the age at onset of Huntington's disease. PMID- 10463358 TI - Immunological effects of in vivo interferon-beta1b treatment in ten patients with multiple sclerosis: a 1-year follow-up. AB - Ten patients with multiple sclerosis and treated with interferon-beta1b (IFN beta1b) were followed-up for 1 year with quantitation of serum VCAM-1 and ICAM-1 levels, mean fluorescence intensity of HLA-DR, VLA-4, CD11a, and CD18 on peripheral blood monocytes and lymphocytes, and adhesion of peripheral blood monocytes and CD45+ cells on endothelial cell monolayers. Adhesion molecule expression and adhesion of peripheral blood monocytes to endothelium were also monitored in healthy controls. No differences in adhesion were detected between MS patients before treatment and healthy controls, while after 1 year a marked decrease in the number of monocytes and mononuclear cells adhering to human umbilical vein endothelial cell monolayers was observed in patients treated with IFN-beta1b. After 1 year of treatment a significant increase in HLA-DR on peripheral blood monocytes was also detected. Our findings regarding lowered adhesion add information to available evidence of the mechanisms of action of IFN beta1b in MS. PMID- 10463360 TI - Cardiovascular autonomic dysfunction in multiple sclerosis: correlation with orthostatic intolerance. AB - Autonomic dysfunction is frequently observed in patients with multiple sclerosis (MS), but clinical studies disagree on the frequency and type of abnormalities in autonomic function tests. Orthostatic dizziness (OD) has been reported in up to 49% of patients, but the pathophysiological mechanisms are poorly understood. This study investigated cardiovascular reflex tests and their association with OD in patients with MS in order to examine the hypothesis that the sympathetic nervous system is specifically involved in these patients. Forty patients with clinically active relapsing-remitting (n = 27) and secondary progressive MS (n = 13), aged 35.0+/-8.5 years, were studied by parasympathetic (heart rate responses to the Valsalva maneuver, deep breathing, and active change in posture) and sympathetic function tests (blood pressure responses to active change in posture and sustained handgrip), and by spectral analysis of heart rate variability during rest and during standing. Results were compared to those obtained in 24 healthy volunteers, aged 29.4+/-7.2 years. A standardized questionnaire was used to evaluate symptoms of orthostatic intolerance. Abnormal responses on at least one cardiovascular reflex test were observed in 40% of MS patients, compared to 17% of the control group, with a statistically significant involvement of the sympathetic vasomotor system. Orthostatic intolerance was reported in 50% of patients (controls: 14%, P<0.006). Subgroup comparison of patients with and without OD suggests that orthostatic intolerance results from impaired sympathetic vasoconstriction. These results provide further evidence that the sympathetic nervous system is involved in patients with MS. PMID- 10463361 TI - Exploring differences between subgroups of multiple sclerosis patients in health related quality of life. AB - Health-related quality of life (HRQoL) questionnaires finding increasing use in medicine, but still only few publications have reported on HRQoL in patients with multiple sclerosis (MS). Because of its progressive and disabling nature, the disease has a considerable effect on HRQoL. To facilitate the interpretation of scores for the heterogeneous MS population, we explored differences between subgroups of MS out-patients (n = 90) on two HRQoL instruments: the Medical Outcome Study Short Form-36 and the Disability and Impact Profile. Three disease related characteristics were studied: severity of MS, time since diagnosis, and MS progression in the past 6 months. The results show that each of these have an effect on one or more aspects of HRQoL. Thus, the longer the disease duration and the more severe and progressive the MS, the lower is the patient's experienced HRQoL. PMID- 10463362 TI - Transcranial Doppler detection of microembolic signals in patients with Behcet's disease. AB - We evaluated the prevalence of microembolic signals (MES) in patients with Behcet's disease (BD). We also attempted to determine the frequency of MES in BD patients with or without neurological involvement. This study enrolled 55 patients fulfilling the diagnostic criteria of International Study Group for BD. Bilateral transcranial Doppler ultrasound of the middle cerebral arteries was performed. MES were identified based on the criteria of International Consensus group on Microembolus Detection. Patients with BD were divided into two groups in respect of the presence of neurological involvement (n = 10) or not (n = 45), and counts of MES in the two were compared with each other and with normal subjects. We found MES in 16 patients (29%) with BD. The frequency was higher in patients with neurological involvement than in those without (80% vs. 17%, P< 0.001). In patients with neurological involvement there was a positive correlation in regression analysis between the prevalence of MES and disease duration (P = 0.025). There was a significantly higher prevalence of MES in BD patients than in control subjects. The frequency of MES was higher in patients with neurological involvement than in those without. TCD detection of MES may allow the recognition of subset of patients at high risk for the appearance of neurological involvement. PMID- 10463363 TI - Phenotypic variation of a new P0 mutation in genetically identical twins. AB - We have identified a new point mutation in the myelin protein zero (P0) gene in two genetically identical twins with a demyelinating neuropathy. The G to A transition at nucleotide position 382 caused an aspartic acid to asparagine substitution in exon 3. Moreover, we found clear clinical differences which were most evident at an early age. These observations suggest that the expression of this P0 mutation may be susceptible to external, non-genetic influences that may act early in the course of the disease to alter the phenotype. PMID- 10463364 TI - Saccades and smooth pursuit in myotonic dystrophy. AB - Reflexive saccades, remembered saccades, antisaccades, fixation and smooth pursuit were recorded in seven subjects with myotonic dystrophy (MD) and seven age-matched controls using the magnetic scleral search coil technique. Neuropsychological performance was assessed using the Wisconsin Card Sort Test and measures of verbal fluency. Subjects with MD showed significantly elevated error rates in the antisaccade and remembered saccade paradigms, consistent with prefrontal dysfunction, and these two measures of distractibility were significantly correlated with each other. Saccadic latencies, square wave jerk frequency, and smooth pursuit peak velocity gain showed no significant difference between the two groups, although the peak velocity of all classes of saccades was significantly reduced in patients with MD. These results extend the findings of previous studies of oculomotor function in MD, and provide novel evidence for a central contribution to abnormalities of eye movements in this condition. PMID- 10463365 TI - Reductions in the bilateral parietal and occipital cerebral blood flow and metabolism in a patient with Marchiafava-Bignami disease. PMID- 10463366 TI - Recurrent brain edema in ornithine-transcarbamylase deficiency. PMID- 10463367 TI - Neurological involvement and quadricuspid aortic valve in a patient with Ehlers Danlos syndrome. PMID- 10463368 TI - Erythropoietin induced visual hallucinations after bone marrow transplantation. PMID- 10463369 TI - Toxic leukencephalopathy following "ecstasy" ingestion. PMID- 10463370 TI - Intrathecal baclofen for intractable spasticity in amyotrophic lateral sclerosis. PMID- 10463371 TI - Pathological yawning (chasm) associated with periodic leg movements in sleep: cure by levodopa. PMID- 10463372 TI - Acquired neuromyotonia: superiority of plasma exchange over high-dose intravenous human immunoglobulin. PMID- 10463373 TI - Avoidance of lithium intoxication: advice based on knowledge about the renal lithium clearance under various circumstances. AB - To avoid lithium intoxications, clinicians should counteract conditions under which lithium clearance changes. If that is not possible, the lithium dosage should be adjusted to the lithium clearance. Lithium clearance is usually 20-30 per cent of the glomerular filtration rate and varies with it, for example during pregnancy and kidney disease. Lithium clearance may also change independently of the glomerular filtration rate. It decreases for example during dehydration (water deficiency) and low sodium intake, pathological conditions with edema formation, treatment with diuretics, anti-hypertensive drugs, or non-steroid anti inflammatory drugs. It increases during treatment with some vasodilating drugs. Precautionary measures against lithium intoxication are suggested for each of these situations. PMID- 10463374 TI - Depressed in-patients respond differently to imipramine and mirtazapine. AB - Tricyclic antidepressants and more recent antidepressants are generally considered to have equivalent efficacy in the treatment of depression. After a previous report of a marked difference in the response to mirtazapine compared to imipramine, we report here an analysis of different symptom clusters. One hundred seven consecutive in-patients with major depression (Diagnostic and Statistical Manual III-R, DSM-III-R) and a Hamilton Rating Scale for Depression (HRS-D) score of 18 points or more were randomly assigned to double-blind treatment. Two and four weeks after predefined blood levels had been obtained, the severity of depression was assessed using the HRS-D. The mean dosages used were 235 mg/day of imipramine and 77 mg/day of mirtazapine, the latter being in excess of the 15-45 mg/day range currently advised. Total HRS-D scores and seven symptom clusters were analyzed in the 85 patients (79%) who were not receiving any co-medication. Imipramine was more effective against the clusters related to core symptoms of depression: "depression and guilt", "retardation", and "melancholia", respectively. Mirtazapine showed a biphasic response with regard to the clusters "sleep" and "anxiety/agitation", respectively, which consisted of a marked response after two weeks of predefined blood level, but with a waning of this effect at four weeks. Imipramine produced a more gradual response on these clusters, which was more pronounced at four weeks than with mirtazapine. Two aspects of the present study could be related to this finding: blood level control resulted in optimal treatment with imipramine but not mirtazapine, and - most importantly - the patients were not receiving any anxiolytic or hypnotic co medication. These findings suggest that mirtazapine may have anxiolytic and sedative properties and fewer antidepressant properties than imipramine in severely depressed in-patients. PMID- 10463375 TI - Activated Borna disease virus in affective disorders. AB - BACKGROUND: Borna disease virus (BDV) is an animal pathogen that causes behavioral changes in animals. Previous studies have found a high prevalence of serum antibodies as well as Borna disease viral antigens (BDVAGs) and RNA in the white blood cells of psychiatric patients, especially those with affective disorders. The present study attempts to offer a better description of the BDVAG cohort using clinical parameters. METHODS: The prevalence of BDVAG was examined in the peripheral mononuclear leukocytes of patients with a major depressive episode. A subgroup of patients underwent further clinical analysis. RESULTS: In this pilot study, at least, there was a significant difference in the prevalence of BDVAG between psychiatric inpatients with a major depressive episode and control individuals. It also appeared that BDVAG is more frequent in patients with recurrent major depression or bipolar disorder than in those with any other psychiatric disorder studied. The number of previous depressive episodes, as well as symptoms involving fatigue and concentration difficulties were positively related to BDVAG. CONCLUSIONS: The high rate of BDVAG, especially in fatigued patients with recurrent major depression or bipolar disorder, may be a nonspecific aspect of immunosuppression. The question remains whether this neurotropic virus may contribute to the pathogenesis of some types of affective disorder. PMID- 10463376 TI - [Opinion of the "Ethical and Just Questions" work group for neuropsychopharmacology and pharmacopsychiatry (AGNP)]. PMID- 10463377 TI - Double-blind, randomized, placebo-controlled clinical trial on the efficacy and tolerability of a physostigmine patch in patients with senile dementia of the Alzheimer type. AB - Owing to the pharmacokinetic properties of physostigmine when administered by conventional routes, long-term cholinergic treatment of Alzheimer's disease is difficult to manage. In order to overcome the problems associated with the oral and intravenous application of physostigmine, and to improve patients' compliance, a transdermal therapeutic system was developed. The efficacy and tolerability of this system were evaluated in a double-blind, randomized, multicenter study comparing patches containing 30 mg and 60 mg physostigmine with a placebo patch. The clinical trial followed the basic principles of the various guidelines on the evaluation of anti-dementia drugs, and included patients with mild to moderate probable Alzheimer's disease. A total of 204 patients with probable Alzheimer's disease were included in the study. Of these, 136 patients were eligible for the according-to-protocol analysis of efficacy, 167 subjects for the intention-to-treat analysis of efficacy, and 181 patients were included in the safety analysis. In contrast to the hypothesis to be tested, the efficacy of physostigmine was not superior to that of placebo after a treatment period of 24 weeks. On the contrary, there was even a slight, but not statistically significant, trend toward a better outcome in the placebo group. Median physostigmine plasma concentrations of approximately 100 pg/ml were measured, showing a high degree of interindividual variability and no linear dose relationship between the 30 mg and 60 mg dosages. Plasma cholinesterase activity was not significantly affected by physostigmine. The physostigmine patch application in doses of 30 mg and 60 mg apparently did not lead to physostigmine plasma concentrations that were sufficient to compensate for cholinergic deficiencies in affected brain areas and produce clinical benefits. Both the drug and the transdermal system were generally well tolerated under the study conditions. Modifications of the patch system may perhaps make it possible to achieve higher physostigmine plasma concentrations, which seem to be required to induce the expected beneficial effects during long-term treatment of Alzheimer's disease. PMID- 10463378 TI - Treatment of alcohol withdrawal syndrome with gabapentin. AB - Four in-patients with moderate alcohol-withdrawal syndromes benefited from treatment with gabapentin administered in an add-on fashion to clomethiazole. In comparison with the amount of clomethiazole required as estimated using a specially developed score during previous detoxifications of these patients at our hospital, gabapentin (400 mg q.i.d.) clearly reduced the amount of clomethiazole needed now Gabapentin, an anticonvulsant with favorable pharmacokinetic properties and tolerability, and with no known risk of dependence, may therefore be a useful new drug in the treatment of alcohol withdrawal. We believe that the potential value of gabapentin in alcohol withdrawal deserves further controlled studies. PMID- 10463379 TI - Genetic determinants of drug-induced agranulocytosis: potential risk of olanzapine? AB - Whether or not olanzapine causes bone marrow toxicity is still a matter of debate. In spite of pre-marketing and post-marketing clinical trials, and although there have been no cases in animals of olanzapine-induced neutropenia or agranulocytosis, the risk of bone marrow toxicity cannot be excluded. The present paper addresses the following questions: what is the potential background of drug induced agranulocytosis? Are there any case reports supporting the view that olanzapine has relevant bone marrow toxicity? What strategies might be helpful in identifying the pathological mechanisms underlying this side effect? PMID- 10463380 TI - Citalopram, a selective serotonin reuptake inhibitor, improves symptoms of Friedreich's ataxia. AB - Two patients (siblings) with Friedreich's ataxia showed improvement in their clinical symptoms--particularly spasticity and cardiac symptoms--after treatment with the selective serotonin reuptake inhibitor citalopram. PMID- 10463381 TI - Papassotiropoulos et al., "The risk of acute suicidality in psychiatric in patients increases with low plasma cholesterol" (Pharmacopsychiatry 32 (1999) 1 4) PMID- 10463382 TI - CYP2E1-dependent benzene toxicity: the role of extrahepatic benzene metabolism. AB - Benzene, a ubiquitous environmental pollutant, is haematotoxic and myelotoxic. As has been shown earlier, cytochrome P450 2E1 (CYP2E1)-dependent metabolism is a prerequisite for the cytotoxic and genotoxic effects of benzene, but which of the benzene metabolites produces toxicity is still unknown. The observed differences between the toxicity of benzene and that of phenol, a major metabolite of benzene, could be explained by alternative hypotheses. That is, whether (1) toxic benzene effects are caused by metabolites not derived from phenol (e.g. benzene epoxide, muconaldehyde). which are formed in the liver and are able to reach the target organ(s); or (2) benzene penetrates into the bone marrow, where local metabolism takes place, whereas phenol does not reach the target tissue because of its polarity. To further investigate hypothesis 2, we used various strains of mice (AKR, B6C3F1, CBA/Ca, CD-1 and C57B1/6), for which different toxic responses have been reported in the haematopoietic system after chronic benzene exposure. In these strains, CYP2E1 expression in bone marrow was investigated and compared with CYP2E1 expression in liver by means of two independent methods. Quantification of CYP2E1-dependent hydroxylation of chlorzoxazone (CLX) by high performance liquid chromatography (HPLC; functional analysis) was used to characterize specific enzymatic activities. Protein identification was performed by Western blotting using CYP2E1-specific antibodies. In liver microsomes of all strains investigated, considerable amounts of CYP2E1-specific protein and correspondingly high CYP2E1 hydroxylase activities could be detected. No significant differences in CYP2E1-dependent enzyme activities were found between the five strains (range of medians, 4.6 12.0 nmol 6-OH-CLX/[mg protein x min]) in hepatic tissue. In the bone marrow, CYP2E1 could also be detected in all strains investigated. However, chlorzoxazone hydroxylase activities were considerably lower (range of medians, 0.2-0.8x10(-3) nmol 6-OH-CLX/[mg protein x min]) compared with those obtained from liver microsomes. No significant (P>0.05) interstrain differences in CYP2E1 expression in liver and/or bone marrow could be observed in the mouse strains investigated. The data obtained thus far from our investigations suggest that strain-specific differences in the tumour response of the haematopoietic system of mice chronically exposed to benzene cannot be explained by differences in either hepatic or in myeloid CYP2E1-dependent metabolism of benzene. PMID- 10463383 TI - Haemoglobin adducts of acrylonitrile and ethylene oxide in acrylonitrile workers, dependent on polymorphisms of the glutathione transferases GSTT1 and GSTM1. AB - Fifty-nine persons with industrial handling of low levels of acrylonitrile (AN) were studied. As part of a medical surveillance programme an extended haemoglobin adduct monitoring [N-(cyanoethyl)valine, CEV; N-(methyl)valine. MV: N (hydroxyethyl)valine, HEV] was performed. Moreover, the genetic states of the polymorphic glutathione transferases GSTM1 and GSTT1 were assayed by polymerase chain reaction (PCR). Repetitive analyses of CEV and MV in subsequent years resulted in comparable values (means, 59.8 and 70.3 microg CEV/1 blood; 6.7 and 6.7 microg MV/1 blood). Hence, the industrial AN exposures were well below current official standards. Monitoring the haemoglobin adduct CEV appears as a suitable means of biomonitoring and medical surveillance under such exposure conditions. There was also no apparent correlation between the CEV and HEV or CEV and MV adduct levels. The MV and HEV values observed represented background levels, which apparently are not related to any occupational chemical exposure. There was no consistent effect of the genetic GSTM1 or GSTT1 state on CEV adduct levels induced by acrylonitrile exposure. Therefore, neither GSTM1 nor GSTT1 appears as a major AN metabolizing isoenzyme in humans. The low and physiological background levels of MV were also not influenced by the genetic GSTM1 state, but the MV adduct levels tended to be higher in GSTT1- individuals compared to GSTT1 + persons. With respect to the background levels of HEV adducts observed, there was no major influence of the GSTM1 state, but GST- individuals displayed adduct levels that were about 1/3 higher than those of GSTT1 + individuals. The coincidence with known differences in rates of background sister chromatid exchange between GSTT1- and GSTT1 + persons suggests that the lower ethylene oxide (EO) detoxification rate in GSTT1- persons, indicated by elevated blood protein hydroxyethyl adduct levels, leads to an increased genotoxic effect of the physiological EO background. PMID- 10463384 TI - Increase in urinary excretion of 6beta-hydroxycortisol in common marmosets as a marker of hepatic CYP3A induction. AB - The ratio of urinary 6beta-hydroxycortisol (6beta-OHF) to free cortisol (F), i.e., the 6beta-OHF/F ratio, has been reported to be a specific marker for human CYP3A induction by in vivo studies of human subjects. In the development of drugs, it is quite beneficial to predict human CYP3A induction in preclinical safety studies using urine samples from experimental animals. We examined the 6beta-OHF/F ratio in urine of common marmosets administered with rifampicin, a potent inducer of CYP3A, to evaluate the usefulness of common marmosets for the prediction of CYP3A induction. Rifampicin was orally administered to three groups of four male common marmosets at doses of 0, 10, and 20 mg/kg per day for 4 days. Amounts of 6beta-OHF and F in urine samples were determined by means of high performance liquid chromatography (HPLC) during the experimental period. One day after the 4th dosing, animals were killed, and P450 contents and P450-catalyzed. 7-alkoxycoumarin O-dealkylase (ACD) activities in the liver were measured. Western blot analysis of liver microsomes was also performed using anti-rat P450 (CYP1A1, 2B1/2, 3A, and 4A) antibodies. The results indicated elevations in the 6beta-OHF/F ratios that were dependent on both the dosing period and dose levels adopted. The ratios on day 4 reached 4.7- and 5.3-fold the pre-administration values in the 10 and 20 mg/kg per day groups, respectively. P450 contents and ACD activities were also elevated in all of the groups. Western blot analysis showed specific increases in the protein which cross-reacts with anti-rat CYP3A antibody in all of the groups. Furthermore, the 6beta-OHF/F ratio was well correlated with the CYP3A contents in liver (r = 0.906). These results indicated that increase in urinary excretion of 6beta-OHF is a specific marker of the induction of hepatic CYP3A in common marmosets just as in humans. Consequently, the present study suggested that human CYP3A induction elicited by chemical agents can be predicted in common marmosets by measuring the urinary excretion of 6beta-OHF. PMID- 10463385 TI - Constitutive and induced expression by pyridine and beta-naphthoflavone of rat CYP1A is sexually dimorphic. AB - Adult male and female Sprague-Dawley rats were compared in terms of the constitutive levels and inducibility of CYP1A1 and CYP1A2 (CYP1A) in lung, kidney, and liver. CYP1A were induced by i.p. treatment with pyridine (75 mg/kg per day) or beta-naphthoflavone (betaNF; 25 mg/kg per day) for two consecutive days and analyzed catalytically (via O-dealkylation of resorufin ethers), at the protein level (by Western blot analysis) and at the mRNA level (by Northern blot analysis). In untreated rats. CYP1A1 protein and its mRNA were detectable only in the lung and kidney of females but not males, whereas CYP1A2 protein and its mRNA were detectable only in the liver in either gender. Pyridine treatment upregulated CYP1A1 mRNA and its protein in the lung, kidney and liver in female rats, and upregulated the mRNA but not the protein in the lung and liver in male rats. Conversely, pyridine induced both CYP1A2 mRNA and protein in the liver in female rats, whereas it induced the protein but not its mRNA in the liver in male rats. No gender difference was observed in the plasma elimination rate of administered pyridine. BetaNF, in contrast to pyridine, induced CYP1A proteins, activities, and mRNA to higher levels in male than in female rats. The results show that the constitutive as well as inducible expression of CYP1A is sexually dimorphic in the Sprague-Dawley rat, with females being more responsive than males to induction by pyridine but with males being more responsive than females to induction by betaNF. The findings support the involvement of different mechanisms in CYP1A induction by pyridine and betaNF. PMID- 10463386 TI - Toxicokinetics of p-tert-octylphenol in female DA/Han rats after single i.v. and oral application. AB - Female DA/Han rats were administered p-tert-octylphenol [OP; p-(1,1,3,3 tetramethylbutyl)-phenol], either intravenously (5 mg/kg body wt.) or orally by gavage (50 or 200 mg/kg body wt.). After i.v. administration the blood concentration-time curve of OP was fitted to a tri-exponential model, resulting in a final half-life (gamma-phase) of 36.1 h. This contrasts to much more rapid eliminations previously reported in male Wistar rats. The oral bioavailability of 50 mg/kg OP was 12.3% and of 200 mg/kg 8.4%. The higher dose (200 mg kg) was absorbed slower than the smaller dose, probably due to low solubility of OP in aqueous media. Maximal OP blood levels in female DA/Han rats receiving 50 and 200 mg OP/kg body wt, were 4.5 and 3 times higher than previously reported in male Wistar rats. The blood concentration-time curves after oral administration of OP to female DA/Han rats revealed pronounced interindividual differences, indicating extensive enterohepatic circulation of OP in this rat strain. In contrast to male Wistar rats, after application of high doses of OP to female DA/Han rats the compound was not completely eliminated within 48 h: under these conditions some bioaccumulation might therefore occur. The experimental toxicokinetics of OP appears as a relevant subject to be integrated into extrapolation of toxicological data, from in vitro to in vivo, and into systems of risk assessment of endocrine modulating activity which are currently being developed. PMID- 10463387 TI - Cadmium-induced calcium release and prostaglandin E2 production in neonatal mouse calvaria are dependent on cox-2 induction and protein kinase C activation. AB - The mechanisms by which cadmium (Cd) causes skeletal impairment have not been fully clarified. Release of calcium from neonatal mouse calvaria in organ culture is stimulated by submicromolar concentrations of Cd, an effect that is associated with increased production of prostaglandin E2 (PGE2). The prostaglandin synthesising enzyme cyclooxygenase (cox) exists in two forms, one constitutive (cox-1) and the other inducible (cox-2). Cox-2 can be induced by mitogenic stimuli and inflammatory cytokines, such as parathyroid hormone (PTH), interleukin-1alpha and tumour necrosis factor-alpha. Cd potently activates protein kinase C (PKC). which in turn induces cox-2 production in several cell types. Our aim was to determine whether Cd-induced Ca release and PGE2 production in neonatal mouse calvaria involve induction of cox-2 and, if so, to ascertain whether that effect is mediated by activation of PKC. Cd dose-dependently stimulated Ca release from cultured neonatal mouse calvaria, with a maximal effect at 0.4-0.8 microM. Different sensitivity was observed to Cd-induced Ca release between two breeds of mice suggesting that the susceptibility to Cd may be genetically determined. Dexamethasone (10 microM) added to the culture medium abolished the Ca releasing effect of Cd, an effect not overcome by addition of arachidonic acid (10 microM). The cox-2-selective inhibitors NS-398 and DFU and the less selective inhibitor meloxicam, potently impeded Cd-induced Ca release (IC50 of 1 nM, 41 nM and 7 nM, respectively) and calvarial production of PGE2. Cd induced and phorbol 12-myristate 13-acetate (PMA; 20 nM)-induced Ca release was inhibited by the PKC inhibitor calphostin C (0.5 microM) and by NS-398. The effects of PMA and Cd on Ca release were not additive, suggesting that both operated via the PKC pathway. We suggest that Cd-induced Ca release from neonatal mouse calvaria in culture depends on induction of cox-2 that occurs via the PKC signalling pathway. PMID- 10463388 TI - Rat model for renal effects of 2-alkoxyalcohols and their acetates. AB - Male Wistar rats were given ethanediol (9.4 g/l), 2-ethoxyethylacetate (5.4 g/l), 2-butoxyethylacetate (2.9 g/l) and 1,2-propanediol (40 g/l) respectively in their drinking water for 2 weeks. Urine was collected during the last 24 h of the exposure. There was a marked increase in the oxalic acid excretion by the rats given ethanediol while rats given the alkoxyacetates excreted large amounts of ethoxyacetic and butoxyacetic acid, respectively. While not increased compared with controls, the excretion of oxalic acid by the latter group of rats was correlated to the excretion of the respective alkoxyacetic acids. The ammonia and glycosaminoglycan excretion was also smaller than that of controls. The urinary activity of succinate dehydrogenase was decreased in rats given the alkoxyacetates but not in animals exposed to ethanediol or propanediol. The data show that oxalic acid is actually a minor metabolite of the alkoxyacetates while the biochemical effects in kidney are associated more with the alkoxyacetic acid load. Alkoxyacetic acids seem to be inhibitors of renal succinate dehydrogenase, which may account for the decreased ammonia and glycosaminoglycan excretion. PMID- 10463389 TI - Urinary antigens as markers of papillary toxicity. II: Application of monoclonal antibodies for the determination of papillary antigens in rat urine. AB - We have previously reported the preparation of monoclonal antibodies specific for antigens localized in the rat renal papilla. Three of the monoclonal antibodies reacting with antigens localized in papillary and cortical collecting duct epithelia were selected for the development of enzyme-linked immunosorbent assay (ELISA)-type assays. The papillary antigens ('PapA') determined in these tests were designated PapAl (applying the monoclonal antibody PapX 5C10), PapA2 (applying the monoclonal antibody PapX 12F6), and PapA3 (applying the monoclonal antibody PapXI 3C7). Using these assays antigen excretion was determined in the urine of rats. Depending on the test compound used. the application route, and the dose, the observed antigen release patterns differed. Whereas after a single intraperitoneal application of 2-bromoethanamine or of propyleneimine an increased release of PapA1 but not of the two other antigens was observed oral application of bromoethanamine had minor effects. In contrast, both a single intraperitoneal application or repeated oral applications of indomethacin resulted in an increased release of all the three antigens. Daily application of ipsapirone in the diet or in drinking water resulted in significantly elevated urinary release of PapAl which increased incrementally for the duration of the application. Release of PapA2 and PapA3 was not affected and remained in the normal range. These results show that with the tests developed changes in the rat renal papilla caused by xenobiotics can be detected early by urinary analysis and monitored during follow-up studies. Moreover. the different antigen release patterns obtained after application of the different compounds suggest a possible differing mode of action. PMID- 10463390 TI - Glutathione transferase alpha as a marker for tubular damage after trichloroethylene exposure. AB - To investigate possible persistent nephrotoxic effects of trichloroethylene (TRI), a retrospective study was carried out on 39 workers exposed to high levels of TRI from 1956 to 1975. Total protein levels in urine, as well as serum and urine creatinine and serum urea were unchanged in comparison with the control. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was applied to differentiate between tubular and/or glomerular dysfunction. Urinary excretion of alpha-1-microglobulin and glutathione transferase (GST) alpha, as markers of proximal tubular damage, were correlated with the SDS-PAGE patterns of urinary proteins both in the TRI exposed and the control group. GST alpha was found in elevated concentrations in the urine of the TRI-exposed workers. No increase of urinary GST alpha was observed in the control group, even when alpha-1 microglobulin was elevated as a result of non-toxic damage. Both in the control and exposed groups, GST pi, a marker of distal tubular damage, was in the normal range. The results show that chronic exposure to high doses of TRI causes persistent changes to the proximal tubular system of the kidney and that GST alpha excretion into the urine is a marker well suited for quantitation of the extent of renal damage. PMID- 10463391 TI - Glutamine synthetase activity in rat urine as sensitive marker to detect S3 segment-specific injury of proximal tubule induced by xenobiotics. AB - The possibility of detecting segment-specific injury of the proximal tubule by means of urinary enzymes was investigated in rats. Urinary glutamine synthetase, an enzyme exclusively localized in the S3 segment, and N-acetyl-beta-D glucosaminidase, prevalently a S1-S2, but S3 enzyme also, were determined after single treatment with 100 mg/kg body wt. of hexachloro-1:3-butadiene (HCBD; i.p.), toxic for the S3 segment, or 25 mg/kg body wt. of potassium dichromate (s.c.), toxic for the S1-S2 segments. Excretion of total urinary proteins was also measured. In addition, a dose-response relationship was determined between three doses (50, 100 and 200 mg/kg body wt.) of HCBD and glutamine synthetase activity in urine. Glutamine synthetase activity, measured according to a new assay for urine based on modification of methods developed for organs, increased in the urine only when the S3 segment of the proximal tubule was damaged, as demonstrated by histological findings of the kidneys. HCBD caused early excretion of the enzyme related to the necrosis of the S3 segment, whereas potassium dichromate caused a slight increase only when the resulting lesion to this segment (vacuolization) began to develop. On the contrary, N-acetyl-beta-D glucosaminidase activity showed the peak of excretion 24 and 34 h after treatment with HCBD or potassium dichromate, respectively, according to the histological findings of necrosis of the S3 segment (the former) and vacuolization of the S1 S2 segments (the latter). Excretion of total urinary proteins reached the peak 24 h (HCBD) and 48 h (potassium dichromate) after treatment. HCBD at 200 mg/kg body wt, caused a peak of glutamine synthetase activity in urine 10 h after injection, whereas the peak caused by doses of 50 and 100 mg/kg body wt. occurred 24 h following treatment. The peak of enzyme activity in urine significantly increased with the dose. The results suggest that the measurement of urinary activity of S3 segment-specific enzyme as glutamine synthetase allows us to detect early S3 segment-specific injury of the proximal tubule. In addition, the method for urinary enzyme activity appears sensitive, simple and fast. PMID- 10463392 TI - Protective effects of capillarisin on tert-butylhydroperoxide-induced oxidative damage in rat primary hepatocytes. AB - Capillarisin (Cap), a main constituent of Artemisia capillaris (Compositae), was studied for its antioxidant bioactivity. In the preliminary study, Cap expressed a antioxidant property by its capacity for quenching the free radicals of 1,1 diphenyl-2-picrylhydrazyl (DPPH). This antioxidant bioactivity of Cap was investigated further using a model of t-butylhydroperoxide (t-BHP)-induced cytotoxicity and genotoxicity in rat primary hepatocytes. Results presented here demonstrate that Cap, at concentrations of 0.01-1.00 mg/ml, significantly decreased the leakage of lactate dehydrogenase (LDH) and alanine aminotransferase (ALT) and the formation of malondialdehyde (MDA) induced by 30 min treatment of t BHP (1.5 mM) in primary cultured rat hepatocytes. Cap also attenuated the t-BHP induced diminution of glutathione (GSH) and high level of DNA repaired synthesis. These results lead to speculation that Cap presents inhibitory effects against t BHP-caused cytotoxicity and genotoxicity in rat primary hepatocyte cultures at least via two distinct pathways, stabilizing the GSH system and quenching free radicals. PMID- 10463394 TI - Doxorubicin induces male germ cell apoptosis in rats. AB - To clarify whether apoptosis is involved in doxorubicin (DXR)-induced testicular toxicity and to identify the target germ cell type, adult Sprague-Dawley rats were treated with a single intravenous dose of DXR (8 or 12 mg/kg) and euthanized at 3, 6, 12, 24, and 48 h subsequently. Histologically, germ cell degeneration was first found 6 h after dosing in meiotically dividing spermatocytes and early round spermatids of seminiferous tubules at stage 1, and subsequently observed in spermatogonia at stages I-VI showing ultrastructural characteristics of apoptosis. Coincident with the appearance of morphological changes, degenerating germ cells were shown to be undergoing apoptosis as revealed by in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The frequency of TUNEL-labeled germ cells increased in a stage- and cell type specific manner, the peak of frequency gradually progressing from stage I of seminiferous tubules to later stages with time after dosing, suggesting that the damaged germ cells, especially spermatogonia, gradually underwent the processes leading to apoptosis. DNA laddering on gel electrophoresis was apparent 24 and 48 h after dosing. The results demonstrate that apoptosis plays an important role in the induction of testicular toxicity caused by DXR with meiotically dividing spermatocytes and type A and intermediate spermatogonia as highly vulnerable target cells. PMID- 10463393 TI - Increased levels of nitrogen oxides and lipid peroxidation in the rat brain after soman-induced seizures. AB - We have investigated the effect of soman-induced seizures on rat brain levels of nitrogen oxides (NOx) and lipid peroxidation (LPO) 30 min and 24 h after intoxication. Following administration of soman (90 microg/kg s.c.), acetylcholinesterase activity was reduced to <10% of control after 30 min, whereas some de novo synthesis had occurred after 24 h. Significant increases in the LPO products malondialdehyde (MDA) and (E)-4-hydroxy-2-nonenal (4-HNE) were seen in the cortex, hippocampus, striatum, thalamus and medulla-pons 30 min after administration. A significant increase in the brain NOx levels, suggesting an increase in NO production, was seen in the cortex after 30 min and in the hippocampus and the striatum after 24 h. No significant changes were observed in cerebellum. These data suggest the possibility that free radical reactions may be a primary cause of neuronal degeneration after soman intoxication. PMID- 10463395 TI - Exposure to epichlorohydrin and dimethylformamide, glutathione S-transferases and sister chromatid exchange frequencies in peripheral lymphocytes. AB - Workers in epoxy resin, synthetic leather, and printed circuit board manufacturing plants are exposed to epichlorohydrin (ECH), or dimethylformamide (DMF), or both. ECH, an alkylating agent, has been shown to cause malignancy in animals, but its genotoxicity in humans is unclear. DMF is a well-known hepatotoxic chemical, although evidence of its genotoxicity in humans is also limited. In this study, we examined the effects of exposure to ECH and DMF on sister chromatid exchange (SCE) in plant workers, in order to examine the genotoxicity of these two agents. Because the genotoxicity of certain agents can be modulated by metabolic traits, we also investigated influence of the glutathione S-transferase (GST) micro (GST M1) and GST theta (GST T1) genes on the genotoxicity of ECH and DMF. A total of 85 male plant workers were included in this study. The subjects were divided into five exposure groups, based on their job titles and the airborne ECH and DMF concentrations in their areas of work. A questionnaire was administered to obtain detailed occupational, smoking, alcohol consumption, and medication histories. Standardized cytogenetic methods were used to determine the frequency of sister chromatid exchange (SCE) in peripheral blood lymphocytes. GST M and GST T1 genotypes were identified using polymerase chain reaction (PCR). In analysis, smoking was significantly associated with increased SCE frequency (P<0.01). Workers with high ECH exposure also had significantly higher SCE frequencies than those with low or no ECH exposure (P<0.05). However, DMF exposure was not associated with SCE frequency. The GST M1 null genotype was also found to be associated with an increased SCE frequency (P = 0.06). We conclude that ECH exposure may be associated with genetic toxicity and that DMF does not appear to be genotoxic. PMID- 10463396 TI - Level and distribution of progesterone in bovine milk in relation to storage in the mammary gland. AB - The study investigated the effect of the place of storage of milk in the mammary gland on progesterone concentrations in whole milk, skim milk and milk fat. Skim milk, milk fat and whole milk progesterone concentrations were lower (P < 0.05) in milk fractions obtained from the cisternal part of the mammary gland compared to those in the milk fractions from the alveoli. Mean milk fat concentrations did not mirror the changes in the mean skim milk, milk fat and whole milk progesterone concentrations. After administration of oxytocin, milk fat concentrations rose significantly (P < 0.01). At the same time, skim milk and milk fat progesterone concentrations remained unchanged (P > 0.05), compared to those in the milk fractions of alveolar origin, obtained before oxytocin administration. Skim milk and whole milk progesterone concentrations were higher (P < 0.01) in composite milk and in milk samples collected 1 h after milking, compared to concentrations in the milk samples collected before morning milking and at 3, 5, 7 and 9 h after milking. The results suggest that defatted milk, milk fat and whole milk progesterone concentrations were affected by the place of storage of the milk in the mammary gland, and that this effect is independent of milk fat content. Time of milk sampling, not the milk fat concentration, in relation to time of milking, was a critical factor in determining skim milk, milk fat and whole milk progesterone. The study also revealed that the concentrations of the other milk components, somatic cell count, lactose and protein were affected by the place of storage of milk in the mammary gland. PMID- 10463397 TI - Influence of grazing management on the seasonal change in testicular morphology in Corriedale rams. AB - The present study was conducted: (a) to determine the degree of seasonal variation in testis stereology in Corriedale rams between autumn and winter; (b) to test the hypothesis that testis stereology of Corriedale rams grazing native pastures during autumn and winter would differ from those of Corriedale rams grazing sown pastures and supplemented with grain during the same period; and (c) to determine whether Sertoli cell numbers differ in adult rams between the breeding season (autumn) and the following non-breeding season (winter). Twenty experimental animals were studied. Six rams (autumn control group, C-A) that had been grazing on native pasture (stocking rate = 2-3 animals ha(-1)) were castrated at the beginning of the experiment (March, early autumn). Seven rams (winter control group, C-W) continued to graze on native pasture at the same stocking rate until the end of the experiment (August, late winter). Another seven rams (treated group, T) grazed on improved pasture (stocking rate = 1-2 animals ha(-1)) and were supplemented with 1 kg grain ram(-1) day(-1) until the end of the experiment. Live weight, scrotal circumference, serum testosterone concentration and selected testicular stereological parameters were measured. The treatment did not impede the winter reduction in testicular activity and reduced its magnitude slightly (group T) compared with controls (group C-W). Sertoli cell numbers were higher in autumn (group C-A) than in winter, both on native (group C W) and sown pastures (group T). Diminishing Sertoli cell numbers between autumn and the following winter suggest the occurrence of that Sertoli cell death during this period. The results indicate that, although the reproductive activity of Corriedale rams is moderately seasonal, a restricted change in grazing and grain supplementation can only modify it to a limited extent. PMID- 10463398 TI - Studies on the measurement of phospholipase A2 (PLA2) and PLA2 inhibitor activities in ram semen. AB - Extraction with Tris-citrate or Tris-NaCl-EGTA improved the yield of phospholipase A2 (PLA2) from ram semen by 40-50 fold over the previously recommended method of extraction by dilute (0.18 N) sulphuric acid. The enzyme activity in the citrate extract deteriorated more rapidly than in Tris-NaCl-EGTA. The semen PLA2 activity was optimum at pH 8.0, heat sensitive at 70 degrees C for 30 min, activated by Ca2+ (although approximately 60% activity was also found in the absence of calcium) and did not exist as a pro-enzyme. The semen PLA2 activity was equally distributed among the sperm and seminal plasma (SP) components of ram semen. However, the low levels of PLA2 activity in the SP of vasectomised rams tend to suggest that PLA2 in the SP fraction may have originated from testicular or epididymal secretions or leakage, from sperm. PLA, in sperm exists as a large molecular weight aggregate, whereas in SP it is present as a smaller aggregate. In addition to PLA2, semen also contained PLA2 inhibitor activities. Inhibition was observed against PLA2s from bee venom, pig pancreas and oviductal extracts. The inhibitory activity is presumed to be due to a large molecular weight protein as the inhibitor activity was not extracted in a chloroform:methanol (2:1; v/v) mixture, it was non-dialysable, precipitated by 10% trichloroacetic acid and destroyed by proteases. The inhibitor activity was distributed in various molecular weight fractions of sperm, SP and SP from vasectomised rams. PMID- 10463399 TI - Analysis of serum and seminal plasma after feeding ochratoxin A with breeding boars. AB - The aim of the experiment was to investigate whether or not ochratoxin A (OA) can be detected in seminal plasma after feeding the toxin in five and 10 times of the human tolerable daily intake with breeding boars and how toxin profiles of serum and seminal plasma correspond to each other. In addition to that, the effect of the toxin challenge on motility and longevity of boar semen was also evaluated. OA from samples was analyzed by microplate ELISA. Percentage of progressive motility of spermatozoa was determined initially and after 24, 48, 96, 120 and 144 h of storage. OA appeared in serum and seminal plasma shortly after toxin application had started. Significant reduction of initial motility and impaired longevity was observed after toxin withdrawal. These findings suggest that OA might have the potential to affect sperm production and semen quality of boars, but further research is required to elucidate whether OA exerts direct effect on germinal epithelium or disturbs sperm cell maturation only. PMID- 10463400 TI - Development of pig embryos reconstructed by microinjection of cultured fetal fibroblast cells into in vitro matured oocytes. AB - Nuclear transfer as originally developed for use in amphibians involved microinjecting a nucleus directly into the cytoplasm of the oocyte. A major mammalian modification has been to use cell fusion to introduce the nucleus. Here we report using a microinjection method to introduce small and medium sized fibroblast cells into mature oocytes. Small cells were more likely to result in nuclear formation (30%) than larger cells (15%; P = 0.013). Small, confluent and serum starved cells resulted in nuclear formation more often (P < 0.048) than did cycling cells. The rate of nuclear formation was not dependent upon the media, (NCSU-23 or TL-Hepes without calcium) nor upon the duration of exposure to the media (1 h to 4 h) after microinjection but before activation. While such treatments did not have an effect on nuclear formation, treatment of parthenogenetically activated oocytes with calcium-free TL-Hepes reduced the percentage of blastocysts (P = 0.068. 11.2% vs. 18.3%) and increased the percentage of morula stage embryos (P = 0.007; 27.6% vs. 15.7%) as compared with culture in NCSU. Finally, small confluent cells were used for nuclear transfer and resulted in two presumptive blastocyst stage embryos [2/128 injected or 2/38 (5.3%) successful injections]. These results show that presumptive blastocyst stage embryos can result from microinjection of fibroblast cells to enucleated oocytes and thus may provide a method to create transgenic knockout animals. PMID- 10463401 TI - Correlation between LH response to challenges with GNRH and naloxone during lactation, and LH secretion and follicular development after weaning in the sows. AB - The aim of this study was of establishing a correlation between endogenous LH secretion and the magnitude of the LH response to challenges with GnRH and the opioid antagonist naloxone during lactation, and between these characteristics and LH secretion and follicular development after weaning. Sows (n = 9) were sampled for 6 h at day 2 post-partum, for 12 h on day 26 of lactation and for 6 h immediately after weaning at day 27 of lactation. Four hours after the beginning of sampling at day 26 of lactation all sows were injected with 2 mg/kg i.v. of naloxone hydrochloride and 5 h later with 100 microg/sow of GnRH. Follicular development was studied in all sows at slaughter the day after weaning. There was an effect of time (sampling period; P < 0.001) on mean plasma LH, with an increase (P < 0.05) in LH the day after weaning compared to mean LH concentrations during lactation. Naloxone and GnRH treatment both increased (P < 0.05) mean LH concentrations. A positive relationship (r = 0.58, P < 0.01) between mean plasma LH after GnRH and after weaning was established. Although there were differences (P < 0.001) between sows in follicular fluid volume, there were no correlations between mean follicular fluid volume and mean LH concentrations after GnRH or after weaning. These data indicate that the LH response to GnRH during late lactation could be useful predictor of LH activity after weaning. However, none of the measures of endogenous or induced LH secretion were associated with differences in ovarian follicular size after weaning. Direct evidence is therefore still needed for a functional link between differences in LH in lactation and differences in fertility after weaning. PMID- 10463402 TI - Epidemiology of the pseudo-exfoliation syndrome. AB - 1) Pseudo-exfoliation syndrome (PE syndrome) is a condition with worldwide distribution. 2) Marked geographical variations have been demonstrated. 3) Environmental factors may be of etiological significance for the development of PE syndrome. 4) The PE syndrome's ability to promote glaucoma may vary from one area to another. 5) There seems to be a trend towards lower debut age of PE syndrome at lower latitudes. 6) More prevalence studies from different parts of the world are needed. A standardized set-up of such works would facilitate comparison of the results, and so some guidelines have been proposed. PMID- 10463403 TI - Diabetic maculopathy caused by disturbances in retinal vasomotion. A new hypothesis. PMID- 10463404 TI - Reliability of the drop test for the lacrimal drainage function. AB - PURPOSE: The purpose of this study was to investigate the reliability of the drop test. METHODS: The repeatability of the test was studied with the same and three different examiners. The influence of the blink rate was investigated by recording the blink frequency in 63 subjects. The degree of reflex lacrimation during the test was assessed in ten patients. The effect of different test solutions was investigated in ten test subjects. RESULTS: There was no significant difference when the test was repeated either by the same or different examiners. The reflex lacrimation during the drop test was not significant. There was no correlation between drop test result and blink rate. A moderate increase in the viscosity of the test solution affected the lacrimal drainage. CONCLUSION: The drop test is a reliable test for the lacrimal drainage function. PMID- 10463405 TI - Effect of Octreotide on experimental corneal neovascularization. AB - PURPOSE: To examine the ability of subcutaneously administered Octreotide ( a long acting somatostatin analoque) to serve as an inhibitory agent for corneal neovascularization in eyes of Wistar Albino rats. METHODS: Neovascular growth into the corneas of all the animals was induced by silver nitrate cauterization. Half of the animals which were randomly selected for the Octreotide group received 30 micrograms systemic Octreotide for 7 days. The treatment was initiated on the same day as chemical cauterization. The rest of the animals (control group) received no treatment. Slit lamp and histopathologic examination of the corneas of both groups were performed at the end of the study period. RESULTS: It was observed that the corneal neovascularization and histopatologic scores of the Octreotide group were significantly lower than those of the control group (p < 0.001, p < 0.01). CONCLUSION: Systemic administration of Octreotide inhibits the corneal neovascular response in a rat model. PMID- 10463406 TI - Magnification changes in specular microscopy after corneal refractive surgery. AB - PURPOSE: To describe the effect of corneal refractive surgery on the magnification of a contact and a non-contact specular microscope. METHOD: The magnification of a contact specular microscope (Konan Clinical Specular Microscope) and a non-contact specular microscope (Topcon SP-1000) was experimentally and theoretically studied as a function of anterior corneal refractive power and central corneal thickness. RESULTS: The magnification of the contact and non-contact specular microscope was found to decrease slightly with decreasing central corneal thickness. In addition, the magnification of the non contact specular microscope decreased slightly with decreasing anterior corneal refractive power. CONCLUSION: As the preoperative and postoperative measuring conditions are different in patients undergoing corneal refractive surgery a correction for magnification changes is necessary when small changes in endothelial cell density are looked for. PMID- 10463407 TI - Ultrasound biomicroscopy examination of intraocular lens haptic position after phacoemulsification with continuous curvilinear capsulorhexis and extracapsular cataract extraction with linear capsulotomy. AB - PURPOSE: Intraocular lens (IOL) haptic position in 35 eyes that had undergone cataract surgery was examined with ultrasound biomicroscopy (UBM). METHODS: In a prospective randomized study the patients were operated by phacoemulsification using continuous curvilinear capsulorhexis (CCC) (group I) or by extracapsular cataract extraction (ECCE) using linear capsulotomy (group II). Ultrasound biomicroscopy was used to localize both haptics of the implanted intraocular lenses and to measure anterior chamber depth (ACD), iris thickness and anterior chamber angle. The inflammatory reaction in the anterior chamber was assessed with laser flare photometry. Slit lamp examination was performed. RESULTS: Both IOL haptics were found in the lens capsule in all 18 eyes in group I. In group II one of the haptics was located out of the capsule in 7 of 17 eyes (41%). The difference is statistically significant (p=0.01). Postoperatively mean ACD measured with the UBM was 4.06+/-0.30 mm in group I and 3.64+/-0.24 mm in group II (p=0.00025). CONCLUSION: The UBM examinations indicate that phacoemulsification with continuous curvilinear capsulorhexis is a more reliable technique than ECCE with linear capsulotomy to achieve implantation of the intraocular lens haptics in the capsular bag. PMID- 10463408 TI - Survival of glaucoma patients. AB - PURPOSE: To investigate the survival of patients with capsular or simple glaucoma compared with that of the common population, with particular attention to the impact of sex and use of acetazolamide (Diamox). METHODS: The 30 year survival of 1147 patients with capsular or simple glaucoma who were finally hospitalized at the Eye Department, Rikshospitalet, Oslo, from 1961 to 1970, are analysed, using log rank tests. The time varying impacts of sex and acetazolamide on survival are also studied using a regression model. RESULTS: There was a significant increased mortality for patients with acetazolamide, and for men also those not using it. The observed mortality for men was initially lower than the average Norwegian population, but later the mortality increased more rapidly in the glaucoma group. This may be explained by a selection of the healthiest patients to Rikshospitalet, and actually indicates that the excess mortality is even higher than calculated here. CONCLUSION: The analysis of data indicated increased mortality for glaucoma patients when the disease had lasted for some time. This was especially pronounced for men using acetazolamide. A similar study from a period when acetazolamide was not in common use and an analysis of causes of death is also asked for. PMID- 10463409 TI - Detection of shallow detachments in central serous chorioretinopathy. AB - PURPOSE: To study the correlation between symptoms, clinical findings, fundus photographic morphology, and optical coherence tomography in patients with symptoms of central serous chorioretinopathy, but ambiguous biomicroscopic findings and no late-phase fluorescein angiographic leakage. METHODS: Biomicroscopic slit-lamp examination, greyscale digital fundus photography in red free illumination, fundus fluorescein angiography, optical coherence tomography, and focal retinal argon laser photocoagulation. Seven consecutive patients aged 32-69 years, of whom four received focal retinal photocoagulation treatment. RESULTS: All patients demonstrated a shallow serous detachment on optical coherence tomography. After treatment the detachment resolved, as did the relative scotoma, the prolonged dark adaptation, and the dyschromatopsia. Micropsia was markedly reduced in all, but not completely eliminated in two of the patients. CONCLUSION: Patients with central serous chorioretinopathy may have shallow foveal detachments that can only be detected by optical coherence tomography, whereas clinical and angiographic signs of detachment may be missing. Classical symptoms coupled with a normal or only mildly reduced visual acuity are highly indicative of the presence of a serous neuroretinal detachment, but fluorescein angiography is necessary to establish the diagnosis and the target for treatment. PMID- 10463410 TI - Evaluation of tear secretion and tear film stability in pseudoexfoliation syndrome. AB - PURPOSE: In pseudoexfoliation syndrome, a material of unknown composition is deposited at the anterior structures of the eye and at periorbital tissues such as the conjunctiva, where goblet cells and accessory lacrimal glands are located. In this study, the possible effect of pseudoexfoliation on tear secretion and tear film stability was investigated. METHODS: Tear film break-up time and Schirmer tests were performed on 108 eyes of 57 patients with pseudoexfoliation and 120 eyes of 60 normal subjects. Results were statistically analyzed. RESULTS: Both Schirmer test and break-up time scores were significantly lower in the pseudoexfoliative patients (average values 10.13 mm and 6.91 sec, respectively) compared to the control group (average values 12.75 mm and 12.75 sec, respectively). The difference was more prominent in males. CONCLUSION: The lower scores in Schirmer and break-up time tests in the pseudoexfoliation group could be attributed to conjunctival involvement. Pseudoexfoliative patients could be more prone to developing xerophthalmia, especially if they are treated with beta blockers. PMID- 10463411 TI - Pachometry before and after vitrectomy with silicone oil injection. AB - PURPOSE: To evaluate differences in central corneal thickness (CCT) before and after vitrectomy with injection of silicone oil as vitreous substitution material in order to determine whether silicone oil causes more frequent and/or pronounced corneal decompensation than other substitution materials. METHODS: CCT was measured by ultrasonic pachometry in 42 eyes. 25 eyes received injection of silicone oil and 17 eyes C3F8, atmospheric air or saline. Measurements were performed 24 hours pre operatively, during anaesthesia pre operatively and 24 hours, 48 hours and five months post operatively. RESULTS: A progressive central corneal thickening could be demonstrated in both groups. Eyes which received silicone oil showed maximum CCT increase of 0.083 mm (14.1%) 48 hours post operatively. At follow-up CCT had decreased to baseline value in both groups. Silicone oil did not induce CCT changes that were significantly different from those induced by other substitution materials. CONCLUSION: Silicone oil injection in detachment surgery induces reversible corneal decompensation to the same degree as other substitution materials within the first five months post operatively. PMID- 10463412 TI - Long-term survival after ruthenium plaque radiotherapy for uveal melanoma. A meta analysis of studies including 1,066 patients. AB - PURPOSE: Most case series of patients managed by brachytherapy for uveal melanoma are small and survival data show considerable inter-study variation. The aim of this study was to summarise similarly structured case series by meta-analysis. METHODS: A systematic review generated 5 similarly structured case series including survival data for 1,066 patients treated by ruthenium plaque radiotherapy for uveal melanoma. After assessing the inter-study clinical heterogeneity, data were weighed for study size and pooled. RESULTS: Patient and radiotherapy characteristics were largely homogenous, but tumour size varied considerably between studies. The 5-year melanoma-related mortality rate was 6% for small and medium tumours (T1/T2) and 26% for large (T3) tumours. The 5-year and 10-year melanoma-related mortality rates for a balanced set of tumours with small, medium and large tumours being present in similar proportions were 14% and 22%, respectively. CONCLUSION: Case series of patients with uveal melanoma managed by brachytherapy may be pooled to increase sample size and study power. The present estimate of survival following ruthenium plaque radiotherapy compares favourably with previously summarised data of survival after enucleation for similarly sized tumours. PMID- 10463413 TI - The ophthalmology of internal carotid artery dissection. AB - PURPOSE: Carotid dissection is an important cause of cerebral and retinal ischemic symptoms, especially in young adults. This article presents a patient material and also includes a review of the ophthalmologic signs and symptoms of carotid dissection. MATERIAL AND METHODS: Twenty-eight patients with spontaneous dissection of the extracranial internal carotid artery dissection underwent a neuro-ophthalmological examination. RESULTS: Twenty-three patients had oculosympathetic paresis. Two experienced transient monocular blindness; in one the episodes were provoked by sitting up from a supine position. One patient presented with a monocular visual field defect, due to posterior ischemic neuropathy, and two others with homonym hemianopia. Diplopia, caused by an incomplete VIth cranial nerve palsy was recognised in one. CONCLUSION: All but one patient presented with detectable ophthalmologic symptoms or signs. Very often the ophthalmologist is the first medical contact for patients with internal carotid artery dissection. The condition is serious, and a prompt evaluation and treatment is indicated to prevent irreversible sequelae. PMID- 10463414 TI - Age related macular degeneration in monozygotic twins and their spouses in Iceland. AB - PURPOSE: To examine the importance of genetic factors in age-related macular degeneration by using a twin study to compare the concordance of age-related macular degeneration in monozygotic twin pairs and their spouses. METHODS: This was a prospective study that included 50 twin pairs and 47 spouses. Zygosity was determined by genetic laboratory testing. Macular findings were graded based on the grading system used by the Macular Photocoagulation Study Group and the International ARM Epidemiological Study Group. RESULTS: The concordance of age related macular degeneration was 90% in monozygotic twin pairs which significantly exceeded that of twin/spouse pairs (70.2%); p=0.0279. In the nine pairs that were concordant, fundus appearance and visual impairment were similar. Environmental factors and medical history were essentially the same in the twin pairs. CONCLUSION: The statistically significant higher concordance of age related macular degeneration in monozygotic twins compared to their spouses strongly suggests the importance of genetic factors. PMID- 10463415 TI - The comparison of cyclopentolate and atropine in patients with refractive accommodative esotropia by means of retinoscopy, autorefractometry and biometric lens thickness. AB - PURPOSE: In this study, we aimed to compare the cycloplegic effect of cyclopentolate HCI 1% and atropine sulphate 1% in patients with refractive accommodative esotropia by means of retinoscopy, autorefractometer and the measurement of lens thickness by biometry. METHODS: Thirty-two patients with refractive accommodative esotropia aged from 5 to 10 (mean 6.8+/-1.4), had a deviation under 10 prism diopters, and underwent retinoscopic, autorefractometric and biometric study in dry and wet conditions. RESULTS: The retinoscopic, autorefractometric and biometric findings of the right eye were 5.10+/-1.21 diopter (D), 5.03+/-1.20 D, 3.43+/-0.16 mm with cyclopentolate, and 5.2+/-1.2 D, 5.2+/-1.2 D, 3.4+/-0.1 mm with atropine. In the left eye, the measurements were 5.2+/-1.4 D, 5.1+/-1.4 D, 3.5+/-0.2 mm with cyclopentolate, and 5.3+/-1.2 D, 5.20+/-1.3 D, 3.4+/-0.2 mm with atropine, respectively. When these obtained data were compared by the Student's t test no statistical significance was found (p > 0.05). CONCLUSIONS: We suggest that the cyclopentolate cycloplegia applied to the patients with refractive accommodative esotropia is sufficient to produce good cycloplegia, with an effect similar to atropine. PMID- 10463416 TI - Retinopathy in diabetic patients aged 15-50 years in the county of Umea, Sweden. AB - PURPOSE: To examine the prevalence of diabetic retinopathy and its relation to certain risk factors (glycosylated hemoglobin, blood pressure, serum creatinine, proteinuria, smoking) in a population-based study on a specific age-group of patients with diabetes mellitus in the county of Umea, Sweden. METHODS: All diabetic patients aged 15-50 years living in the county of Umea were invited to the study. A standard clinical and eye examination was performed, and seven-field stereoscopic photographs were taken of each eye. Blood and urine samples were collected. Univariate and multivariate statistical analyses were performed. RESULTS: Of the eligible 395 patients 285 (91%) participated in the study. 285 patients (79%) had diabetes mellitus type 1, 71 (20%) subjects had diabetes mellitus type 2, and 3 patients (1%) had secondary diabetes. In the statistical analysis performed on patients with type 1 diabetes mellitus, duration, presence of hypertension, systolic blood pressure, plasma glucose, glycosylated hemoglobin, serum creatinine, proteinuria and smoking all were significantly related to increasing degree of retinopathy when a univariate model was applied. However, when a multivariate analysis was performed only duration, proteinuria, glycosylated hemoglobin and male gender were statistically significantly associated with severeness of retinopathy. CONCLUSION: Increased duration of diabetes, inadequate metabolic control as measured by glycosylated hemoglobin, proteinuria and male gender are factors that are associated with a higher incidence of diabetic retinopathy in diabetes mellitus type 1. PMID- 10463417 TI - Persistent corneal oedema following ICCE. Influence of IOL type. AB - PURPOSE: To examine the frequency of persistent corneal oedema following intracapsular cataract extraction (ICCE). To examine whether the type of intraocular lens has any influence on persistent corneal oedema following ICCE. METHODS: A retrospective study of 1041 consecutive ICCE operations followed for at least eight years. The cumulative incidence of persistent corneal oedema was calculated for the whole group and compared to the incidence in subgroups defined by type of intraocular lens. RESULTS: The cumulative incidence of persistent corneal oedema following ICCE was 2.3% for the whole group. The highest incidence (10.3%) was found in the group of patients, who received a 3-M style 77 anterior chamber lens while the lowest incidence (1.0%) were found among patients receiving a Choyce anterior chamber lens. Compared to this, the patients who received no intraocular lens showed a frequency of persistent corneal oedema of 1.4%. Among the patients who developed persistent corneal oedema the number of re operations was higher (27.3%) compared to the patients who did not develop persistent corneal oedema (4.4%). The frequency of preoperative dystrophy was the same whether or not the patients developed persistent corneal oedema. There were no difference in gender, age or other pre-operative data between the two groups. CONCLUSION: After ICCE 2.3% developed persistent corneal oedema. There was a significant difference between the frequencies of persistent corneal oedema depending on the type of intraocular lens from 1.0% in eyes receiving a Choyce anterior chamber lens to 10.3% in eyes receiving a semi-flexible 3-M style 77 anterior chamber lens. PMID- 10463418 TI - Contrast sensitivity function in pseudophakics and aphakics. AB - PURPOSE: To study the effect of posterior chamber intraocular lenses and aphakic spectacles on contrast sensitivity. METHODS: Contrast sensitivity was evaluated in 20 patients in each group of pseudophakics with post chamber IOL (group I), aphakics with spectacle correction (group II), and age and sex-matched normal subjects (group III) using the Pelli-Robson chart. RESULTS: The mean values of log contrast sensitivity in pseudophakics (1.665+/-0.105) and aphakes with spectacle correction (1.5075+/-0.1) were found to be statistically significantly low (t: 5.186, p < 0.001; t: 11.302, p < 0.001, respectively) as compared to the mean value of normal phakic subjects (1.8075+/-0.0576). Further, mean values of log contrast sensitivity in aphakes with spectacles correction were also found to be statistically significantly low (t: 4.727, p < 0.001) when compared to that in pseudophakes. CONCLUSION: From observations of the present study, it can be concluded that posterior chamber IOL implantation offers an added advantage of higher contrast sensitivity, over and above the well documented advantages of increased field of vision, negligible effect on image size and elimination of prismatic effect and spherical aberration of thick glasses. PMID- 10463419 TI - Indocyanine green videoangiography and color Doppler imaging in Behcet's disease. AB - PURPOSE: The purpose of this study was to examine the status of the retina and the choroid, and to evaluate blood flow velocity in the central retinal (CRA) and ophthalmic artery in Behcet's disease. METHODS: Digital fluorescein and indocyanine green (ICG) videoangiography were performed in 53 eyes and color Doppler ultrasonography in 61 eyes of 31 Behcet patients, between December 1993 January 1996. RESULTS: Fluorescein angiography revealed no vascular leakage in 7, and diffuse retinal and/or disc leakage due to vasculitis in 46 eyes. ICG angiography was normal in 14 eyes while it revealed hypo or hyperfluorescence in the remaining. For the CRA, peak systolic and end diastolic velocities were significantly lower in Behcet patients than in control cases. In the Behcet group, Doppler indices did not differ significantly in eyes which had choroidal abnormalities and those that did not, according to ICG angiography. CONCLUSION: In Behcet's disease, some abnormalities in the choroid may be seen. The disease appears to be associated with decreased mean flow velocities in CRA. PMID- 10463420 TI - Fine needle aspiration biopsy in selecting treatment for inconclusive intraocular disease. AB - PURPOSE: To evaluate the fine needle aspiration biopsy (FNAB) as a diagnostic tool in cases where it was impossible to make a definitive diagnosis with noninvasive techniques. METHOD: 80 consecutive patients with inconclusive diagnoses were examined by FNAB prior to decision of treatment. Biopsies were performed through a transscleral route in 50 eyes, an anterior chamber route in 16 eyes and a transvitreal approach in 14 eyes. The consequences of FNAB were analysed retrospectively. RESULTS: FNAB confirmed malignancy in 59 eyes. Inconclusive material was obtained from 5 eyes judged clinically to be malignant disorders. One melanoma was misinterpreted as being a metastasis. In 47.5% of our patients this procedure altered the therapeutic plan and 25 patients were spared enucleation. The biopsy material was correctly diagnosed as benign in 16 cases. CONCLUSION: In eyes where the diagnosis remained uncertain after non-invasive tests, FNAB gave important information which greatly influenced our choice of treatment. FNAB contained sufficient tissue elements for cytological diagnosis in 77 eyes. Cytopathological interpretation failed once in relation to tumour type. The procedure of FNAB can be recommended for use in ambiguous tumour cases of the eye. Probably it should only be used in tumour centres with adequate cytology service. PMID- 10463421 TI - An unusual case: bilateral orbital varices. AB - PURPOSE: To present a rare case of bilateral orbital varices. METHODS: An 18-year old man showing bilateral orbital masses on magnetic resonance (MR) imaging was examined with color Doppler ultrasonography (US), computed tomography (CT) and digital subtraction venography. RESULTS: The presenting symptoms of the patient were pain and fullness in both orbits induced by bending forward. Ocular examination was normal with the exception of a two millimeter proptosis of both eyes during Valsalva maneuver. MR imaging demonstrated bilateral retrobulbar masses, but was not diagnostic. The comparison of CT images obtained before and after Valsalva maneuver revealed the diagnosis of orbital varices. Color Doppler US and orbital venography demonstrated a large venous connection between the lesions and systemic circulation. CONCLUSION: Clinical presentation of orbital varices is unusual. Different radiological methods may be necessary for the confirmation of the diagnosis and demonstration of the anatomic and the dynamic features of the lesions. PMID- 10463422 TI - A case of congenital orbital cyst originating from the common sheath of superior rectus and levator palpebrae superioris muscles. AB - PURPOSE: To assess the clinical and histopathological features of a case of congenital orbital cyst originating from the common sheath of the superior rectus and levator palpebrae superior muscles. METHODS: Preoperative, peroperative and postoperative characteristics of the described clinical condition in a 37-year old woman is documented. RESULTS: The cyst was found to be related with the common sheath of the superior rectus and levator muscles during the surgical intervention and histopathological examination revealed a cystic structure with a lining resembling that of conjunctiva. CONCLUSION: Our findings support the proposal by Rose & O'Donnell who first reported four cases in 1995 suggesting a new group of congenital orbital cysts distinct from dermoid, epidermoid or conjunctival cysts. PMID- 10463423 TI - Clinical features of Goldmann-Favre syndrome. AB - A 21-year-old woman complained of progressive loss of visual acuity. She had also had night blindness since she was ten years old. At the eye examination, the vitreous was found to be degenerated in both eyes. The fundus findings were a large retinoschisis in the right macula, edema resembling retinoschisis in the left macula and annular degenerative changes in the midperiphery. ERG and dark adaption were abnormal. This vitreoretinal degeneration was diagnosed as Goldmann Favre syndrome. PMID- 10463424 TI - Ciliary body schwannoma. AB - PURPOSE: Intraocular schwannomas are very rare, benign, peripheral nerve neoplasms. The authors report a case of ciliary body schwannoma. METHOD: A 39 year-old Korean woman presented with slowly decreasing visual acuity and proptosis of the right eye for 4 years. At the time of her visit, the eyeball protruded and deviated laterally. She could not sense light with her right eye. We enucleated the eyeball. The enucleated eyeball with tumor was examined histopathologically, immunohistochemically, and under electron microscope. RESULTS: Microscopic examination revealed spindle cells and characteristic Antoni type A and B areas with Verocay bodies. Immunohistochemical study showed tumor cell expressions of S-100 and vimentin, but other immunohistochemical studies were negative. Electron microscopic examination demonstrated Luse body. CONCLUSIONS: We diagnosed the tumor as a schwannoma arising from the ciliary body on the basis of the above mentioned results. Schwannomas are very rare intraocular neoplasms, but they are benign so we should differentiate from other intraocular neoplasms. PMID- 10463425 TI - Loss of iridolenticular contact in eyes with exfoliation syndrome may protect against glaucoma. AB - PURPOSE: To provide evidence for the hypothesis that dynamic iridolenticular contact predisposes to the development of glaucoma in exfoliation syndrome (XFS). METHODS: We present four patients with bilateral XFS and unilateral exfoliation glaucoma (XFG) whose normotensive eyes had suffered traumatic loss of dynamic iridolenticular contact. RESULTS: All 4 patients had bilateral XFS and developed XFG only in the untraumatized eyes. One patient had loss of iridolenticular contact in the traumatized eye, two had a nonreactive pupil, and one had had intracapsular cataract extraction at age 28. CONCLUSIONS: Loss of dynamic iridolenticular contact may help to protect against development of glaucoma in eyes with XFS. PMID- 10463427 TI - Congenital retinal macrovessels: a "low visual acuity" case report with a 14-year follow-up. AB - We report a rare case of congenital retinal macrovessel with decreased visual acuity, with a 14-year follow-up. Both the clinical findings and the visual acuity remained unchanged throughout the follow-up period. PMID- 10463426 TI - Control of cytomegalovirus retinitis after combination antiretroviral therapy. AB - PURPOSE: To report on AIDS patients having combination antiretroviral therapy whose cytomegalovirus (CMV) retinitis remained inactive after discontinuation of anti-CMV maintenance therapy. METHODS: We describe the course of CMV-retinitis in 3 patients with AIDS after initiation of combination antiretroviral therapy. RESULTS: After cessation of anti-CMV therapy no relapse of CMV-retinitis has been observed for up to 18 months. Two of the patients developed new CMV-retinitis in the first months after initiation of combination therapy, nevertheless after further improvement of immunological parameters retinitis remained stable without anti-CMV therapy. CONCLUSION: The sustained immunological effects of combination therapy are possibly sufficient enough to provide protection against CMV retinitis. PMID- 10463428 TI - Spontaneous absorption of a cataractous lens. AB - A case of spontaneous absorption of a cataractous lens is presented. A 37-year old woman with Down's syndrome presented with bilateral cataracts. On follow-up, the cataract in her right eye was found to be absorbed with no secondary uveitis or glaucoma. Surgical capsulotomy was performed on the remnant anterior and posterior capsules. Such absorption is known in juveniles and in hypermature cataract, but is rare in adults in the absence of injury or inflammation. Absorption occurred over a period of one year. PMID- 10463429 TI - Hyperferritinaemia-cataract syndrome. AB - The first family with hyperferritinaemia-cataract syndrome in Scandinavia is described. We discuss the etiology and point at this diagnosis as a differential diagnosis in case of hereditary cataract. PMID- 10463430 TI - Silicone intubation for obstruction of the nasolacrimal duct in adults. PMID- 10463431 TI - Epiretinal membrane in association with Takayasu disease. PMID- 10463432 TI - A case of isolated acute optic neuritis. PMID- 10463433 TI - Relaxing action of sodium nitroprusside independent of membrane potential in the CCh-induced contracture of the guinea pig stomach muscle. AB - Carbachol (CCh, 10(-6) M) induced biphasic contraction of longitudinal muscle of the guinea pig stomach, consisting of rapid phasic contraction and contracture. The contracture was almost completely inhibited by sodium nitroprusside (SNP, 10( 6) M) and S-nitroso-N-acetyl penicillamine (SNAP, 10(-6) M). A membrane permeable analogue of cyclic GMP, 8Br-cGMP (10(-4) M), also inhibited the CCh induced contracture. Although a heme site inhibitor of nitric oxide-sensitive guanylyl cyclase, 1-H-[1, 2, 4] oxadiazolo-[4, 3 a]quinoxalin-1-one (ODQ; 10(-6) M), reduced the inhibitory action of SNP, it did not affect the inhibitory action of 8Br-cGMP, indicating that the effect of SNP was developed via cyclic GMP production in the presence of D600. Charybdotoxin (10(-7) M), an inhibitor of Ca2+ activated K+ channel, did not influence on the CCh induced contracture. On the other hand, CCh induced a depolarization of the longitudinal muscle cell membrane (from -60 mV to -45 mV) in the presence of 10(-6) M D600, but SNP did not affect the depolarization. These results suggest that in the presence of D600 SNP induces relaxation of CCh induced contracture of the longitudinal muscle of the guinea pig stomach via cyclic GMP but not membrane potential dependent mechanism. PMID- 10463434 TI - Effects of superoxide dismutase on the acetylcholine-induced relaxation response in cholesterol-fed and streptozotocin-induced diabetic mice. AB - High concentration of acetylcholine (ACh) caused a rapid and long lasting relaxation response in age-matched controls, whereas this response was significantly weaker in streptozotocin (STZ)-diabetic and cholesterol fed mice. The levels of basal and ACh-stimulated cyclic GMP in the aorta was also significantly smaller in STZ-diabetic and cholesterol-fed mice. The attenuated relaxation responses to ACh in both STZ-diabetic and cholesterol-fed mice were ameliorated by the chronic administration of cholestyramine. A prior incubation of aortic strips with superoxide dismutase (SOD, 60 U/ml) improved the recovery phase of the relaxation of diabetic aorta after single administration of ACh, whereas SOD had no effects on ACh-induced relaxation of aortic strips from cholesterol fed mice. These results suggest that superoxide anion may be responsible for an impairment of endothelium-dependent relaxation of aorta from STZ induced diabetic mice. It is further suggested that impairment of endothelium dependent relaxation in STZ-diabetic and cholesterol fed mice may be caused by different mechanisms. PMID- 10463435 TI - Effects of Dai-kenchu-to on intestinal obstruction following laparotomy. AB - To confirm the usefulness of Dai-kenchu-to for intestinal obstruction, investigation of the effects of Dai-kenchu-to on postoperative intestinal adhesion was conducted. Repeated administrations of Dai-kenchu-to (100 or 300 mg/kg) significantly inhibited the formation of intestinal obstruction. Motor disturbance and inflammation are thought to be involved in the etiology of intestinal adhesion. A single treatment of Dai-kenchu-to (300 mg/kg) significantly reduce intestinal transit time in postoperative ileus and chemically induced ileus. Dai-kenchu-to (10(-4) g/ml) significantly inhibited COX 2 activity. These results suggest that Dai-kenchu-to prevents postoperative intestinal adhesion by gastroprokinetic and anti inflammatic effects. Dai-kenchu to thus demonstrates positive effect on postoperative ileus. PMID- 10463436 TI - Pharmacological studies of the effect of Dai-kenchu-to on spontaneous contraction of isolated rabbit jejunum. AB - We studied the effects of Dai-kenchu to on the spontaneous contraction in isolated rabbit jejunum. Dai-kenchu-to (10(-3) g/ml) increased jejunal contraction, such as phasic like contraction and contractile amplitude. Zanthoxyli Fructus (2x10(-4) g/ml) exhibited an action identical to that of Dai kenchu-to. While Zingiberis Siccatum Rhizoma (5x10(-4) g/ml) continuously decreased the amplitude of contraction. Ginseng Radix (3x10(-4) g/ml) and Saccharum Granorum (8x10(-3) g/ml) had no effect on spontaneous contraction. Dai kenchu-to and Zanthoxyli Fructus reversed the decrease of contraction produced by atropine. However, phasic like contraction induced in the absence of atropine was antagonized by atropine. Dai-kenchu-to and Zingiberis Siccatum Rhizoma further decreased spontaneous contraction in the presence of tetrodotoxin. It was clarified that Dai-kenchu-to possesses gastroprokinetic effect, and Zanthoxyli Fructus mainly contributed to this effect. It was suggested that the cholinergic and non cholinergic nervous systems were involved in increasing intestinal motility. It was also suggested that Dai-kenchu-to acted on multiple points of the intestine, and actions at these points might intensify to improve ileus. PMID- 10463437 TI - Heat resistance of Escherichia coli O157:H7 in apple juice. AB - The objective was to determine the effect of cider composition on the heat resistance of Escherichia coli O157:H7. The average D52 value in a model Empire apple juice was 18 min with a z value of 4.8 degrees C. Increasing the Brix from 11.8 to 16.5 degrees had no effect on thermal resistance, while increasing L malic acid from 0.2 to 0.8%, or reducing the pH from 4.4 to 3.6 sensitized the cells to heat. The greatest effect on heat resistance was afforded by the preservatives benzoic and sorbic acids: D50 values in ciders containing 1,000 mg/l were 5.2 min in the presence of sorbic acid and only 0.64 min in the presence of benzoic acid. Commercial apple juice concentrates yielded lower numbers of survivors than single-strength juices even though their higher sugar concentrations of about 46 degrees Brix increased heat resistance. PMID- 10463438 TI - Heat resistance of Escherichia coli O157:H7 in a nutrient medium and in ground beef patties as influenced by storage and holding temperatures. AB - Stationary-phase cultures of Escherichia coli O157:H7 were inoculated into tryptic soy broth, sealed in vials, and stored at -18 degrees C for 1, 8, and 15 days, or 3 or 15 degrees C for 3, 6, and 9 h. Thermal resistance was determined at 55 degrees C. Each storage treatment was repeated with additional holding at 23 or 30 degrees C for 1, 2, 3, or 4 h prior to heating to simulate potential temperature abuse during handling. Cultures under treatments enabling the growth of E. coli O157:H7 were generally more heat sensitive than those held at temperatures which restricted growth or enabled growth to stationary phase. Cultures stored frozen (-18 degrees C) without holding at elevated temperatures had greater heat resistance than those stored under refrigeration (3 degrees C) or at 15 degrees C. Subsequent holding of frozen cultures at 23 or 30 degrees C resulted in a decrease in heat resistance. To determine whether these responses would be observed under typical commercial preparation procedures, ground beef patties were inoculated with E. coli O157:H7 and stored at 3 or 15 degrees C for 9 h or at -18 degrees C for 8 d and then held at 21 or 30 degrees C for 0 or 4 h. Patties were grilled to an internal temperature of 54.4 degrees C (130 degrees F), 62.8 degrees C (145 degrees F), or 68.3 degrees C (155 degrees F). Cultures were most resistant in frozen patties, while cultures in patties stored at 15 degrees C were the most heat sensitive. Holding patties at 21 or 30 degrees C prior to grilling resulted in increased sensitivity. Storage and holding temperatures similar to those encountered in food service may influence the ability of E. coli O157:H7 to survive heat treatments. PMID- 10463439 TI - Comparison of a microtitration plate ELISA with a standard cultural procedure for the detection of Salmonella spp. in chicken. AB - A rapid antibody-capture enzyme-linked immunosorbent assay (ELISA) detecting a wide range of Salmonella serotypes and employing only one culture stage was used to analyze the giblets and body cavity rinsings from frozen chickens. The results from the ELISA were compared with those obtained using a standard cultural procedure in current use in two laboratories, Norwich (N) and Ipswich (I), of the Public Health Laboratory Service (PHLS) in the UK. ELISAs were carried out on the same samples at each of two PHLS laboratories and at the Institute of Food Research with good agreement (94% and 90%). When compared with the cultural method there was 80% and 70% agreement with the ELISA with the PHLS(N) and PHLS(I) samples. The ELISA appeared to have a false-positive rate of 17% (samples from PHLS(N)) but on reculture of the "negative" samples this rate fell to 7%. The false-negative rate for the ELISA was 26% (samples from PHLS(N)) which appeared to be due to insufficient growth of the Salmonella spp. in the single cultural step employed in the ELISA rather than lack of recognition by the antibodies. The problem of false negatives with the cultural method is also discussed. These results are comparable to previously published studies relating immunoassays and the conventional procedure for Salmonella detection when analyzing similar samples. Suggestions are made as to how further increases in ELISA efficiency might be brought about. PMID- 10463440 TI - Metabolic activation and inactivation of metanil yellow and orange II in the Salmonella typhimurium his-reversion assay. AB - Among a variety of experimental protocols used, the combined use of 0.5% dextrose in bottom agar and 1 micromol of flavin mononucleotide (FMN) in preincubation mixtures without fraction S9 mix resulted in the highest level of induced Salmonella typhimurium his reversions with both dyes metanil yellow and orange 11 with strain TA100. Strain TA98 yielded notably lower levels of reversions under the same conditions. The presence of uninduced hamster liver S9 fraction resulted in a weak mutagenic response while Aroclor 1254-induced rat liver S9 fraction resulted in the complete absence of mutagenicity with both strains and with both dyes. PMID- 10463441 TI - Monolaurin and acetic acid inactivation of Listeria monocytogenes attached to stainless steel. AB - Individual and combined antimicrobial effects of monolaurin and acetic acid on Listeria monocytogenes planktonic cells or stainless-steel-adherent cells were determined in order to evaluate cell viability during a 25-min exposure period at 25 degrees C. A 10(7)-colony-forming units (CFU)/ml population of planktonic cells was completely inactivated by the synergistic combination of 1% acetic acid with 50 or 100 microg/ml of monolaurin within 25 or 20 min, respectively. Either compound alone caused partial but incomplete inactivation within the same time periods. A population of 10(5) CFU/cm2 of 1-day adherent cells on stainless steel was completely inactivated within 25 min, but with the highest concentrations of the combined chemicals, i.e., 1% acetic acid and 100 microg/ml of monolaurin. The combined chemical treatment again synergistically produced greater inhibition. A 10(6)-CFU/cm2 population of 7-day adherent cells was not completely inactivated within 25 min of exposure, although counts did decline. The results demonstrate increased resistance of attached L. monocytogenes to acetic acid and monolaurin and show that resistance increased with culture age. Combinations of organic acids and monolaurin might be considered as sanitizers of food contact surfaces, but activities of such combinations are likely to be less than other commonly used sanitizers. PMID- 10463442 TI - Combined effects of packaging atmosphere and lactic acid on growth and survival of Listeria monocytogenes in crayfish tail meat 4 degrees C. AB - The effect of lactic acid on growth and survival of Listeria monocytogenes in crayfish tail meat stored under refrigeration and various gas environments was investigated. Frozen crayfish tail meat was thawed overnight, autoclaved, cooled, and inoculated with approximately 4 log colony-forming units (CFU) of a mixed strain (Scott A and F5027) L. monocytogenes culture per gram of meat. Inoculated samples were blended with 0, 0.5, 1.0, 1.5, or 2.0% lactic acid and packaged under air, vacuum, or modified atmosphere (74.8% CO2, 10.4% O2, and 14.8% N2) and stored at 4 degrees C for 20 days. Results demonstrated that modified atmosphere packaging inhibited the growth of L. monocytogenes more than air and vacuum packaging at 0 and 1% lactic acid. Microbial counts declined steadily in crayfish tail meat treated with 2% lactic acid, with no differences among the packaging atmospheres. The lag phase was extended by 8 days in samples treated with 1% lactic acid and modified atmosphere compared to that in air or vacuum packaging. Overall, the combination of lactic acid and modified atmosphere had the greatest potential to prevent growth of L. monocytogeines. PMID- 10463443 TI - Risk of Clostridium botulinum type E toxin production in blue crab meat packaged in four commercial-type containers. AB - The aim of this investigation was to determine if a risk of Clostridium botulinum growth and toxin production existed in four different packaged crabmeat products. Freshly picked blue crab meat was inoculated with 10(3) to 10(4) spores per g of a mixed pool of four strains of C. botulinum type E (Beluga, Minnesota, G21-5, and 070). The lump crabmeat was packaged in four different packaging containers: (i) 12-oz copolymer polyethylene cups currently used by most crab processors; (ii) 12-oz copolymer polyethylene cups with heat-shrink, tamper-evident low density polypropylene seals; (iii) 8-oz copolymer polyethylene cups with easy open aluminum ends: and (iv) 8-oz copolymer polypropylene cups with integral tamper-evident pull-tabs. The packages were stored at either 4 degrees C for 21 days or 10 degrees C for 15 days. Storage at 10 degrees C was used to simulate temperature abuse. The mouse bioassay was used to detect the presence of C. botulinum toxin. Psychotrophic and anaerobic populations were enumerated and were found to increase with time regardless of packaging type. No botulinum toxin was detected in any of the four packaging types stored at 4 degrees C or 10 degrees C throughout the entire storage period. PMID- 10463444 TI - Potential Bacillus subtilis alpha-amylase-based time-temperature integrators to evaluate pasteurization processes. AB - Thermal inactivation kinetics of Bacillus subtilis alpha-amylase (BSA) in different environmental conditions was studied by performing isothermal experiments. As a response property, residual enzymic activity and residual heat of enzyme deterioration were chosen. A comparison of processing values determined from the read-out of a system with actual integrated processing values revealed the potentials of these systems as time-temperature integrators to be used in the pasteurization domain (temperatures of 70 to 100 degrees C) for target attributes with z-values ranging from 6 to 12 degrees C. PMID- 10463445 TI - Fungicidal effect of 15 disinfectants against 25 fungal contaminants commonly found in bread and cheese manufacturing. AB - Resistance of 19 mold and 6 yeast species to 15 commercial disinfectants was investigated by using a suspension method in which the fungicidal effect and germination time were determined at 20 degrees C. Disinfectants containing 0.5% dodecyldiethylentriaminacetic acid, 10 g of chloramine-T per 1, 2.0% formaldehyde, 0.1% potassium hydroxide, 3.0% hydrogen peroxide, or 0.3% peracetic acid were ineffective as fungicides. The fungicidal effect of quaternary ammonium compounds and chlorine compounds showed great variability between species and among the six isolates of Penicillium roqueforti var. roqueforti tested. The isolates of P roqueforti var. carneum, P. discolor, Aspergillus versicolor, and Eurotium repens examined were resistant to different quaternary ammonium compounds. Conidia and vegetative cells were killed by alcohols, whereas ascospores were resistant. Resistance of ascospores to 70% ethanol increased with age. Both P. roqueforti var. roqueforti and E. repens showed great variability of resistance within isolates of each species. PMID- 10463446 TI - Resistance to aflatoxin contamination in corn as influenced by relative humidity and kernel germination. AB - Kernels of corn population GT-MAS:gk, resistant to aflatoxin B1 production by Aspergillus flavus, and susceptible Pioneer hybrid 3154 were tested for aflatoxin when incubated under different relative humidities (RH). High aflatoxin levels were not detected in either genotype at RH < 91%. Resistance in GT-MAS:gk was consistent across all RH levels (91 to 100%) at which significant aflatoxin accumulation was detected. Aflatoxin levels in GT-MAS:gk averaged about 98% less than those in susceptible Pioneer 3154, which suggests that storage of this or other genotypes with similar resistance mechanisms may be possible under moisture conditions less exacting than are required with susceptible hybrids. Results for fungus growth and sporulation ratings on kernel surfaces were similar to those for aflatoxin levels. When kernels of both genotypes were preincubated 3 days at 100% RH prior to inoculation with A. flavus, germination percentages increased to very high levels compared to those of kernels that were not preincubated. In preincubated kernels aflatoxin levels remained consistently low in GT-MAS:gk but decreased markedly (61%) in Pioneer 3154. When eight susceptible hybrids were evaluated for aflatoxin accumulation in preincubated kernels, seven of these supported significantly lower toxin levels than kernels not subjected to preincubation. Average reduction across hybrids was 83%, and reductions within hybrids ranged from 68 to 96%. Preincubated kernels of one susceptible hybrid (Deltapine G-4666) supported aflatoxin levels comparable to those in resistant GT MAS: gk. Data suggest that an inhibitor of aflatoxin biosynthesis may be induced during kernel germination. Possible mechanisms for embryo effects on resistance to aflatoxin accumulation are discussed. PMID- 10463447 TI - Hazard analysis and critical control point evaluation of school food programs in Bahrain. AB - Hazard analyses were conducted in six food preparation sites and 16 school canteens in the State of Bahrain. Sandwiches made with cheese, meat, eggs, liver, and beef burgers were prepared in small shops or a bakery outside schools. Foods were cooked between 4 and 5 A.M. Time-temperature exposure during cooking was adequate to kill vegetative microbes and their spores, but potential for recontamination existed from the hands of food workers, utensils, and cloths and sponges used for wiping. All foods were left at room temperature before they were transported in vans to schools where they were also kept at room temperature between 17 degrees C and 41 degrees C. Air temperature inside the canteens during this investigation was between 18.5 and 28 degrees C with a relative humidity of 65 to 70%. Hazard analyses, which included observation of operations inside school canteens and sites of food preparation, measuring temperatures, and interviewing workers and consumers (teachers, students) were carried out. Hazards were primarily associated with preparation of foods long before they were consumed, physical touching of products, and holding foods at room temperature after preparation. Holding foods at room temperature would have allowed germination of bacterial spores and multiplication of microbes. Reheating of foods was not practiced. Health promoters must be aware of these hazards and need to educate food workers, administrators, and the public on the methods of prevention. PMID- 10463448 TI - Consumer knowledge of foodborne microbial hazards and food-handling practices. AB - A national telephone survey was conducted of 1,620 randomly selected U.S. residents who spoke English, were at least 18 years old, and resided in households with kitchen facilities. Respondents were interviewed about their recognition of foodborne pathogens, foods at risk for transmitting infection, knowledge of safe food handling, and food-handling practices. One-third of the respondents who prepared meals reported unsafe food hygiene practices: e.g., they did not wash hands or take precautions to prevent cross-contamination from raw meat. Unsafe practices were reported more often by men, adults 18 to 29 years of age, and occasional food preparers than by women, persons 30 years old or older, and frequent food preparers. Respondents who identified a food vehicle for Salmonella spp. were more likely to report washing their hands and cleaning cutting boards after preparing raw meat and poultry. The results raise concerns about consumer food-handling practices. The influence of food safety training, food-handling experience, and age on food-handling practices should be studied further. Awareness of a food vehicle for Salmonella spp., for example, may indicate knowledge of the etiology of foodborne disease that promotes safe food handling. Understanding the factors associated with safe food handling will assist in development of effective safe-food instruction programs. PMID- 10463449 TI - Detection of trichinellosis in pigs by artificial digestion and enzyme immunoassay. AB - The objective of this study was to demonstrate the reliability of current and proposed methods for the inspection of swine and other species for infection with the parasite Trichinella spiralis. Five groups of pigs were infected with doses of 2500, 500, 100, 50, and 20 T. spiralis larvae to establish moderate and low level infections. Pigs were bled periodically during the study for samples to be tested by enzyme immunoassay (EIA). At the conclusion of the study, pigs were slaughtered and tissues collected for analyses of worm burdens and for comparison of digestion testing methods. Comparisons of pooled sample digestion methods were made using inspection methods prescribed by European Union Directives and the USDA, Code of Federal Regulations. Pooled sample digestion testing using 1-g samples was effective for detecting pigs with larval densities of > 10 larvae per gram (LPG) of tissue but only partially effective for pigs with infections of <3 LPG. Pooled sample digestion testing using 5-g samples detected all pigs with infection levels > 1 LPG. The EIA detected all T. spiralis-infected pigs, but did not detect infections in some pigs until 49 days after inoculation. These results demonstrate that the pooled sample digestion method using a 5-g sample size is the most effective inspection method for reducing the risk of human exposure to T. spiralis in pork. PMID- 10463450 TI - Quality characteristics of fresh blue crab meat held at 0 and 4 degrees C in tamper-evident containers. AB - There has been a regulatory movement toward the required use of tamper-evident containers for fresh blue crab meat. North Carolina passed tamper-evident regulations in 1993. Blue crab processors had little information on possible changes in head-space gases, microbial growth, chemical decomposition, sensory quality, or shelf life caused by the new containers. Chemical, microbiological, physical, and sensory changes in fresh crab meat were monitored during 18 days of storage in ice and 13 days of storage refrigerated at 4 degrees C. "Special" blue crab meat, chosen for the study, is the least expensive commercial form of white crab meat. The crab meat was packaged in four retail containers: copolymer polyethylene cups with polyethylene snap-on lids, copolymer polyethylene cups with snap-on polyethylene lids fastened to the cup with heat-shrink low-density polypropylene seals, copolymer polyethylene cans with aluminum easy-open ends, and copolymer polypropylene cups with a tamper-evident pull-tab on the lid. Control samples packaged in industry standard copolymer polyethylene cups maintained higher oxygen levels than meat stored in tamper-evident containers. No consistent differences in quality or shelf life were detected among the containers. Market shelf life was limited to 6 days for meat held at 4 degrees C and 15 days for meat held at 0 degrees C. Sensory quality deteriorated 6 days earlier for crab meat held at 4 degrees C than meat held at 0 degrees C. Collateral work showed that toxin production by Clostridium botulinum neither occurred following 18 days of storage at 4 degrees C nor after 15 days of storage at 10 degrees C. Definite spoilage occurred before any toxin production. The study suggests that blue crab processors can safely use the new tamper-evident packaging, which has little or no effect on product quality or shelf life. Processors may choose appropriate packaging options using price, packaging quality, market appearance, and ease of production as the deciding criteria. PMID- 10463451 TI - Use of a new rapid bioluminescence method for screening organophosphate and N Methylcarbamate insecticides in processed baby foods. AB - An enzyme with high specific affinity for organophosphate and N-methylcarbamate insecticides has been incorporated into a new test for detection of these insecticides at the level of parts per billion (ppb) (commercially available as the Charm Pesticide Test). To measure the extent of insecticide inhibition of the enzyme, a specific bioluminescent substrate is used. The signal is counterproportional to the amount of insecticides. Random sampling of four baby food brands and testing for the cumulative levels of organophosphate and N methylcarbamate insecticides found carbaryl to be the most common residue. Out of the 155 samples tested there were 132 negative samples (85.2%) and 23 suspected positive samples (14.2%). The suspected positive samples were further analyzed by high-performance liquid chromatography (HPLC) and gas chromatography/mass spectrometry (GC/MS). Carbaryl was confirmed in 18 of the samples. One of the samples contained an active metabolite of tetrachlorvinphos and in 3 of the positive samples an insecticide could not be identified by GC/MS. One positive sample was not processed for confirmation due to high fat content. PMID- 10463452 TI - Proteinase inhibition of fish muscle enzymes using legume seed extracts. AB - Seed extracts from indigenous and introduced legumes were prepared and used to search for inhibitors of fish muscle proteinases. Fish flesh extracts were prepared from samples of Merluccius productus (Pacific whiting or merluza) obtained off the Oregon coast and in the Gulf of California, respectively. The proteinase activity in the fish muscle for the Pacific whiting was the highest, followed by parasitized merluza. The lowest proteinase activity was for the nonparasitized merluza. Six out of 12 seed extracts reduced the proteinase activity from the fish flesh by more than 50%. The reduction of enzyme activity was higher for samples of fish flesh extracts from the Gulf of California than for the Oregon samples. Seed extracts also reduced the proteinase activity of commercial serine and cysteine proteinases such as trypsin, chymotrypsin, and papain. This inhibitory capacity was maintained even after heating the seed extracts to 90 degrees C for 15 min. Several seed extracts show promise for use as proteinase inhibitors during production of surimi, the intended commercial product of massive fisheries such as Pacific whiting or merluza. PMID- 10463453 TI - Inhibition of Salmonella typhimurium on agar medium and poultry skin by ultraviolet energy. AB - Ultraviolet radiation (UV) was effective in destroying Salmonella typhimurium on agar plates and poultry skin. Agar plates inoculated with varying numbers of colony-forming units (CFU) of S. typhimurium (1.2 x 10(2) to 1.7 x 10(9) were subjected to different doses of UV light to determine optimal killing. Poultry skin was also inoculated with varying CFU of S. typhimurium per 2 cm2 of skin and subjected to UV light. UV light treatment of inoculated agar plates revealed almost complete elimination (99.9%) of S. typhimurium at 2,000 microW x s x cm( 2). Bacterial reduction was less effective on the surface of poultry skin when a 80.5% reduction in S. typhimurium was obtained at 2,000 microW x s x cm(-2). PMID- 10463454 TI - Cetylpyridinium chloride (CPC) treatment on poultry skin to reduce attached Salmonella. AB - Cetylpyridinium chloride (1-hexadecylpyridinium chloride, CPC) was evaluated for its effectiveness in removing or killing salmonellae attached to poultry skin. Two different treatment methods were used: (i) spraying 0.1% CPC solution at 15 degrees C or 50 degrees C against inoculated skin surface for I min at 138 kPa, and (ii) immersing inoculated skin surface in 0.1% CPC solution at room temperature for either 1 min, 1 min plus 2 min holding without CPC, or 3 min. After rinsing, cells on the skins were enumerated by conventional plating as well as direct counting from scanning electron microscopy (SEM). Compared with controls, CPC spraying reduced the numbers of salmonellae by 0.9 to 1.7 log units (87 to 98%) assayed by the plating method (P < 0.05). SEM gave results similar to plating. Generally 50 degrees C CPC spraying showed greater reduction than 15 degrees C CPC spraying; however, the differences were not always significant. Water spraying at either temperature did not show any reduction compared to nonsprayed skins. In the immersion test, significant differences also were noticed among the control and the three other CPC-immersed groups (P < 0.05) as assayed by plating, ranging from 1.0 to 1.6 log units, which were similar to the CPC spraying results. However, no difference was noticed among the three CPC immersed groups. Direct counting from SEM was not a suitable method for recovering cells in CPC immersion tests because dead cells were still attached to the skin while retaining their intact morphology. On the basis of the amount of CPC used, immersion appears to be more cost-effective than spraying CPC on poultry skin. PMID- 10463455 TI - Detection of staphylococcal enterotoxin H by an enzyme-linked immunosorbent assay. AB - A double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the detection of a newly identified staphylococcal enterotoxin H (SEH). Peroxidase was conjugated to antibodies specific to the enterotoxin. 2,2'Azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid)(ABTS) in hydrogen peroxide solution was used as the enzyme substrate. A standard curve of purified SEH was prepared with concentrations ranging from 1.3 to 50 ng/ml. SEH at levels equal to 2.5 ng/ml and higher were detected by this procedure. Culture supernatant from the growth of selected Staphylococcus aureus strains was analyzed by using the ELISA. SEH was produced by three of 20 strains that produced one identified enterotoxin. Ten of 21 strains, previously shown to produce substances that induced emesis in monkeys but not any known enterotoxins (A through E), were also positive for SEH production. The other 11 strains gave negative results in the ELISA, indicating that other unidentified serological types of enterotoxin exist. PMID- 10463456 TI - Prenatal exposure to the 1957 influenza pandemic and non-affective psychosis in The Netherlands. AB - Second-trimester exposure to the 1957 A2 influenza pandemic is a controversial risk factor for schizophrenia. Two earlier studies of the Dutch psychiatric registry failed to find an increased risk for exposed subjects, but diagnostic misclassification within the spectrum of non-affective psychoses has not yet been ruled out as an explanation for the negative findings. Using an enlarged data-set we examined not only whether second-trimester exposure to the epidemic is associated with an increased risk of schizophrenia (ICD:295), but also whether it is associated with an increased risk of paranoid states (ICD:297) or other non organic psychoses (ICD:298). In this retrospective cohort study the risks of the above-mentioned disorders were compared for those exposed and unexposed to A2 influenza during the second trimester of fetal life. The risks for the exposed subjects were not significantly higher than the risks for the unexposed. The power of the study to detect a significant increase in the risk of schizophrenia was sufficient. If the relative risk of a lifetime hospitalization for schizophrenia for second-trimester exposed subjects (born January-April 1958) is assumed to be 1.3, the power of the study would be 0.97 (alpha=0.05; one-tailed testing). If the relative risk for subjects born five months after the peak of the epidemic (mid-February to mid-March 1958) is assumed to be 1.88, as reported for England and Wales, the power of the study would be close to 1.00. This was the largest study of its kind in Europe: 275 subjects were born in the period January-April 1958 and had a lifetime hospitalization for schizophrenia. This study indicates that there is no relation between second-trimester exposure to the 1957 influenza pandemic and risk of non-affective psychosis in the Dutch population. It adds to a growing body of work which does not support an association between maternal influenza and schizophrenia. PMID- 10463457 TI - Childhood-onset schizophrenia and obstetric complications: a case--control study. AB - Obstetric complications (OCs) may be a risk factor for developing schizophrenia. In a recent study of a meta-analysis, the odds ratio for the development of the disorder in adulthood associated with OCs has been reported to be about 2.0 (i.e., a two-fold increase in risk). However, little attention has been paid to the involvement of OCs in risk of the development of childhood-onset schizophrenia. Therefore, the authors examined the relationship between OCs and childhood-onset schizophrenia. Thirty-three children, aged 8-13 years (average 12.4 years), meeting the DSM-III-R criteria for schizophrenia, were compared with controls (children with anxiety disorder) matched for sex and age. Childhood onset schizophrenics showed significantly greater scores in all of the three measures of OCs according to Parnas et al.'s scale compared with controls. Moreover, those individuals exposed to OCs were 3.5 times as likely to develop schizophrenia as were those without OCs. The risk association between OCs and the disorder was far greater for male than for female schizophrenics. Our results, together with those in previous studies showing the association between OCs and adult-onset schizophrenia, suggest that childhood- and adult-onset schizophrenia may, at least in part, share a common neuropathogenesis. PMID- 10463458 TI - A rating scale for psychotic symptoms (RSPS) part I: theoretical principles and subscale 1: perception symptoms (illusions and hallucinations). AB - The authors present a new rating scale for the psychotic symptoms of schizophrenia and related psychoses. The scale links specific symptoms of psychopathology to dysfunction and overactivity of dopaminergic mechanisms underlying the processes of reward and selective attention. The Rating Scale for Psychotic Symptoms (RSPS) is a 44-item rating instrument with a seven-point severity scale for each item. Psychotic symptoms are classified into three groups: Pathological amplification of mental images (perception symptoms) (subscale 1), Distraction symptoms (including catatonia and passivity experiences) (subscale 2), and Delusions (subscale 3). A dimensional, rather than a categorical, conceptualization of psychosis is assumed. Rating is accomplished through a manual and a semi-structured interview (SSCI-RSPS). In this first of two papers, general issues about the construction of the scale and the derivation of symptom groups are discussed. Dopamine-mediated modification of cortico striatal synapses is seen as being of critical importance in all three groups of symptoms. In this first paper, we present subscale I (perception symptoms), which includes both amplified perceptual images (illusions) and hallucinations. A total of seven illusions and 11 hallucinations are rated as individual items. PMID- 10463459 TI - A rating scale for psychotic symptoms (RSPS): part II: subscale 2: distraction symptoms (catatonia and passivity experiences subscale 3: delusions and semi structured interview (SSCI-RSPS). AB - In the second paper on the Rating Scale for Psychotic Symptoms (RSPS), distraction symptoms (Subscale 2) and delusions (Subscale 3) are described. Subscale 2 includes distraction symptoms, which arise from one of two mechanisms: the symptom may arise either by loss of attentional focus (LAF) due to a competing channel of information, or by the intrusion of the competing channel into the focus of attention [attentional intrusions (AI)]. The symptom classes resulting from loss of attentional focus (LAF) include motor catatonia, negativism, and thought blocking; the attentional intrusions (AI) symptoms rated include three types of passivity experiences (Schneiderian symptoms): (1) thought insertion, (2) movements or action controlled, and (3) speech controlled by an external force. Subscale 3, consisting of delusions, is organized on the basis of content identification. Nineteen types of delusions are rated. Each item of delusional content is rated along three axes (active elaboration, persistence and active extinction) and complexity, and optionally if primary or mixed (i.e. primary with secondary elaborations). The last section of the paper includes the semi-structured interview (SCI-RSPS) for each of the items, as well as guidelines for practical application of the interview. PMID- 10463460 TI - A controlled study of temporal lobe structure volumes and P300 responses in schizophrenic patients with persistent auditory hallucinations. AB - Recent studies of cerebral pathology in patients with schizophrenia have focused on symptomatological and electrophysiological correlates of reduced temporal lobe structure volumes. Volume deficits of the left superior temporal gyrus have been correlated with auditory hallucinations as well as to left-sided P300 amplitude reduction. However, caution is needed to interpret correlational data as evidence of a specific relationship. Therefore, a controlled study was undertaken on schizophrenic patients with and without auditory hallucinations. MRI-defined volumes of the left superior temporal gyrus and other temporal lobe structures were quantified from 3-mm coronal slices in 15 schizophrenic patients with chronic auditory hallucinations (hallucinators), 15 schizophrenic patients without auditory hallucinations (nonhallucinators) and 17 healthy controls. In all subjects a simple oddball paradigm was used to elicit P300 responses at temporal and centro-parietal electrode sites. No evidence was found for volume reductions of temporal lobe structures in the combined patient group compared with controls, or in the hallucinators compared with the nonhallucinators. The patients did show left P300 amplitude reduction compared with controls, particularly in the hallucinator group. Correlations between volumes of left temporal lobe structures and left P300 amplitudes were low and not significant. The results of the present study do not indicate that auditory hallucinations and associated abnormal electrophysiological activity are the consequence of atrophy of localized temporal lobe structures. However, replication in a larger sample of subjects is needed before firm conclusions can be drawn. PMID- 10463461 TI - Motor dysfunction in schizotypal personality disorder. AB - Past research has revealed that schizophrenia is associated with voluntary movement abnormalities, as well as higher rates of involuntary movements. On instrumental motor tasks, patients manifest reduced motor stability, excessive force and more contralateral motor overflow (movement in the non-responding hand). In the present study, an instrumental motor task (manual response forced choice task) was administered to a group of adults with schizotypal personality disorder (SPD) in order to determine whether they show motor deficits similar to those observed in schizophrenia. As predicted, the schizotypal subjects were excessive and more variable in motor force, compared to healthy controls and other personality-disordered subjects. Additionally, the force and variability of the motor responses were positively correlated with ratings of both positive and negative SPD symptoms. Finally, motor overflow and negative symptoms were associated with higher salivary cortisol levels. The pattern of findings is consistent with previous reports linking motor abnormalities and heightened cortisol with schizotypal personality disorder. PMID- 10463462 TI - Effects of fish oil fatty acids on plasma lipids and lipoproteins and oxidant antioxidant imbalance in healthy subjects. AB - The purpose of this study was to investigate the effect of eicosapentaenoic and docosahexaenoic acids on plasma lipids and lipoproteins, lipid peroxidation and antioxidant status in healthy humans. A total of 19 healthy volunteers consumed 6 g/day Maxepa fish oil for 3 weeks (1.8 g n-3 fatty acids/day). At baseline and at day 21, we evaluated plasma lipoproteins, plasma and low-density lipoprotein fatty acids, lipid peroxidation markers (malondialdehyde concentration, low density lipoprotein peroxidation in vitro), and the content of a number of antioxidants (reduced and oxidized glutathione in whole blood, plasma and erythrocyte glutathione peroxidases, plasma vitamin E and beta carotene). Plasma concentrations of total cholesterol, triglycerides, phospholipids, low-density lipoprotein cholesterol and low-density lipoprotein size did not differ significantly after 3 weeks of supplementation. Adding the fish oil to the diet increased the concentration of n-3 very-long-chain polyunsaturated fatty acids and decreased the concentration of n-6 fatty acid and oleic acid in plasma and low-density lipoprotein. Eicosapentaenoic and docosahexaenoic acid supplementation caused elevated values of the high-density lipoprotein cholesterol due to an increment of the high-density lipoprotein 2 fraction and reduced low-density lipoprotein peroxidation rate in vitro. However, we observed an imbalance between oxidizable substrates and antioxidants with an increased lipid peroxidation, whereas the content of reduced glutathione and beta carotene decreased without any variation in vitamin E. Association of antioxidants with n 3 PUFA could prevent lipid peroxidation and enhance the antiatherogenic effects of n-3 polyunsaturated fatty acids. PMID- 10463463 TI - Endothelial degradation of extracellular lyso-phosphatidylcholine. AB - Formation of lysophospholipids, including lyso-phosphatidylcholine (lysoPC), is enhanced during oxidation of low-density lipoprotein, in ischaemic tissue and under inflammatory conditions. Besides being potentially cytotoxic, extracellular lysoPC induces changes in several properties of vascular endothelial cells. These include expression of endothelial adhesion molecules and interference with the endothelial production of nitrogen monoxide, prostacyclin and growth factors. One way of controlling the concentration of extracellular lysoPC is by the action of lysophospholipases, which degrade lysoPC into a free fatty acid and glycerophosphocholine. We therefore tested whether vascular endothelial cells have the ability to degrade extracellular lysoPC. Monolayers of primary cultures of human umbilical vein endothelial cells degraded an average of 84+/-24 nmol lysoPC/10(6) cells/2 h. By comparison, monocytes degraded 9.7+/-3.7 nmol lysoPC/10(6) cells/2 h, and erythrocytes and platelets < 1 nmol lysoPC/10(6) cells/2 h. The ability of endothelial cells to degrade extracellular phospholipids (diacylphosphatidyl choline) was found to be relatively low (9.5+/ 6.4 nmol/10(6) cells/2 h). Triacylglycerol hydrolase activity was just above detection level. In conclusion, endothelial cells seem to degrade extracellular lysoPC effectively. This endothelial property may be important in controlling plasma and tissue levels of extracellular lysoPC as well as in the interaction between lysoPC and the vascular endothelium. PMID- 10463464 TI - Pro-inflammatory cytokines increase the permeability of paracetamol across a human endothelial-smooth muscle cell bilayer model. AB - Human umbilical vein endothelial cells and smooth muscle cells, cultured on either side of fixed, porous supports, were used to study the effect of pro inflammatory cytokines on the transvascular passage of the drug paracetamol. The cellular bilayer effectively retarded the passage of the drug from the "luminal" or endothelial side, to the "tissue" or smooth muscle side of the bilayer over a 30-min period. When the cells were incubated with either IL-1beta (100 ng/l) or TNF-alpha (10 microg/l) for 4 h prior to exposure to paracetamol, the permeability of the bilayer to the drug increased to that of the control inserts without cells. In contrast, the pro-inflammatory cytokines did not affect the electrical resistance of the bilayer, indicating continued tight junctional integrity, or the passage of [3H]-inulin, an indicator of paracellular transport, or the passage of fluorescein, an indicator of passive diffusion across the cells. Together these data indicate the suitability of this syngenetic human cell co-culture model for studying factors affecting the systemic disposition of drug molecules at the level of the vascular wall. The data also indicate that the transport of paracetamol across the blood vessel wall may be greatly enhanced at sites of tissue inflammation in the systemic circulation. PMID- 10463465 TI - Expression of transforming growth factor alpha and epidermal growth factor receptor in human bladder cancer. AB - We analysed the expression of epidermal growth factor receptor (EGFr) and transforming growth factor alpha (TGF-alpha) in human bladder tumours. Tumour biopsies were obtained from 54 patients with primary bladder cancer (18 stage T1 and 36 stage T2-4). The protein and mRNA expression of EGFr and TGF-alpha were quantified by ELISA and competitive RT-PCR, respectively. The EGFr protein level was significantly increased in T2-4 tumours (0.44 x 10(-11); 0.0-27.5 x 10(-11) mol/g) compared with T1 tumours (0.0; 0.0-2.0 x 10(-11) mol/g) (median; range; 2p<0.01). The EGFr protein and mRNA level correlated (Spearman r=0.45, 2p<0.005, n=40). Co-expression of TGF-alpha protein and EGFr protein was significantly associated with muscle invasive tumours (T2-4) (chi-squared=7.9, df=3, p<0.05) and the TGF-alpha protein level correlated significantly with EGFr protein expression (Spearman r=0.56, 2p<0.0001, n=54). While tumour stage correlated with survival, no correlation was observed between survival and the expression of EGFr and/or TGF-alpha. In conclusion, human bladder tumours express both EGFr and TGF alpha. The expression of EGFr and TGF-alpha are closely correlated, and the expression of EGFr and co-expression of EGFr and TGF-alpha correlate with tumour stage. PMID- 10463466 TI - Phospholipase A2 in serum and colonic mucosa in ulcerative colitis. AB - Group II phospholipase A2 is involved in the pathogenesis of various inflammatory diseases and in the host defence against bacteria. The enzyme is expressed in the epithelial cells of colonic mucosa in ulcerative colitis. In this study, we measured the concentration of group II phospholipase A2 in serum and colonic mucosa of patients with ulcerative colitis of different severity and of control patients without any inflammatory disease. The activity of ulcerative colitis was assessed by endoscopy. The concentration of group II phospholipase A2 was measured with an immunoassay. The concentrations of group II phospholipase A2 in serum and colonic mucosa were significantly higher in patients with active and inactive ulcerative colitis than in controls. However, the group II phospholipase A2 levels did not separate patients with different disease activity. The concentration of group II phospholipase A2 in colonic mucosa corresponded with the mucosal inflammatory activity (higher in active colonic areas) intra individually, but not between different patients with ulcerative colitis. Serum group II phospholipase A2 values were above the normal reference range more often than the values of 11 standard laboratory blood tests widely used for the follow up of inflammatory activity in ulcerative colitis. These results indicate that the concentration of group II phospholipase A2 is increased in serum and colonic mucosa of patients with ulcerative colitis. The clinical value of the measurement of group II phospholipase A2 in the follow-up of ulcerative colitis remains to be clarified. PMID- 10463467 TI - Prevalence of the 985A>G mutation in the medium-chain acyl-CoA dehydrogenase (MCAD) gene in Sweden. AB - The prevalence of the 985A>G mutation in the medium-chain acyl-CoA dehydrogenase gene was determined in the Swedish population. A heterozygote frequency of 1:127 was observed. Morbidity data indicate that most of the homozygotes with this mutation are not diagnosed and probably remain asymptomatic. PMID- 10463468 TI - No microheterogenous changes of plasma C-reactive protein found in man during various diseases. AB - Increased levels of the acute phase protein C-reactive protein (CRP) in plasma may indicate severe acute abdominal disease, risk of serious postoperative complications or malignancy; serial measurements may indicate postoperative complications, relapse of intra-abdominal sepsis and complications during acute pancreatitis. The increase in CRP is an unspecific acute phase reaction, however, and low levels do not exclude these conditions. These facts are important obstacles to the clinical routine use of CRP measurements. The aim of this study was to look for possible biochemical microheterogeneity of CRP in single plasma samples from various large groups of patients to overcome these problems. Two hundred-and-twelve patients with acute abdominal diseases, 274 patients with various forms and stages of cancer and 134 patients operated on due to benign diseases, were studied. The biochemical studies included SDS-PAGE, native PAGE and gel filtration for molecular weight determinations, isoelectric focusing and crossed immuno-electrophoresis for electrophoretic mobility studies and Concavalin A and ACA 34 as intermediary gels for possible lectin binding or complexation. Western blot analysis was also used to identify CRP. In summary, however, these more elaborate biochemical methods could not disclose any microheterogneity of CRP in plasma and thus did not add any diagnostic information to the crude levels. PMID- 10463469 TI - Regional blood flow in the calf and plasma endothelin during prolonged orthostasis in humans. AB - This study tested the hypothesis that circulating plasma levels of the vasoconstrictor endothelin increase during prolonged orthostasis in association with changes in both calf regional blood flow, systemic arterial blood pressure and heart rate, and that the 133Xenon wash-out method can be used for prolonged registration of skeletal muscle blood flow. The simultaneous effects of prolonged orthostasis on plasma endothelin, calf skeletal muscle and subcutaneous blood flow, heart rate and arterial blood pressure were investigated in 13 healthy subjects before, during and after 30 min of 50 degree head-up tilt. Blood flow rate was measured by the local 133Xenon wash-out method in the supine position and during 50 degree head-up tilt. The method was evaluated for skeletal muscle for prolonged observation in the supine position in five subjects and compared with blood flow rates measured with plethysmography. Plasma endothelin was unchanged during head-up tilt, despite a maximal reduction in skeletal muscle blood flow rate of 47% (p<0.001) and subcutaneous blood flow rate of 80% (p<0.01) and a maximal increase in heart rate of 15% (p<0.001) and diastolic (12%, p<0.01) blood pressure. The skeletal muscle wash-out curves for 133Xenon were monoexponential from 30 min after injection and at least during the next hour and correlated with total limb blood flow rate assessed simultaneously with plethysmography (r=0.76, p<0.0001). Circulating endothelin does not appear to be in the first line of regulation of calf microcirculation, systemic arterial blood pressure or heart rate during prolonged orthostasis. The 133Xenon wash-out method can be used for prolonged measurement of skeletal muscle blood flow. PMID- 10463470 TI - Variation in recommendations for cancer screening among primary care physicians in New Mexico. AB - The acceptance of age-appropriate cancer screening as an integral part of primary care has grown among physicians over the past decade. We conducted a mailed survey of all primary care physicians in New Mexico in order to better understand their current cancer screening practices. We found a high rate of self-reported screening, particularly for prostate and colorectal cancer. The screening rates were influenced only slightly by the introduction of evidence-based guidelines, with younger physicians and those with university affiliations more likely to follow recommendations. Female physicians and obstetrician-gynecologists endorsed breast and cervical cancer screening among all age groups and were less likely to follow recommendations for less frequent screening in women as they age. Since a physician's practice beliefs influence his/her attitude toward testing, tailoring education by physician specialty may be more effective than using generic messages in encouraging compliance with the most recent evidence-based guidelines. PMID- 10463471 TI - Pretesting Spanish-language educational radio messages to promote timely and complete infant immunization in California. AB - California's Hispanic infants have lower immunization levels than non-Latino white infants, 53.7% versus 65.2%, respectively. Spanish-language radio is an effective mass media venue for imparting information to Latino populations. It has been demonstrated that lack of parental knowledge of infant immunization timing is associated with delayed immunization coverage. In an effort to improve Latino parent knowledge of immunization timing, two Spanish-language radio commercials were developed to be used in conjunction with community-based educational efforts. In order to gage the potential educational impact of the two commercials, they were pretested with a group of low-income Spanish-speaking Latino parents who represented members of the target population for whom the commercials were created. Both commercials were rated favorably by parents, and elicited immunization-specific responses. Although correct recall of the simplified, basic immunization schedule was low, the level of immunization response consistency and overall approval of both commercials appear to support their use as part of Latino infant immunization educational outreach in California. PMID- 10463472 TI - Health behind bars: utilization and evaluation of medical care among jail inmates. AB - Jail and prison inmates experience disproportionately high levels of chronic and acute physical health problems, resulting in increased utilization of health services in correctional institutions. Variations in both health status and health care utilization are likely, although several important factors have been under-researched. Gender, in particular, is presumed to influence health outcomes and use of medical care in correctional facilities. The current study explores the physical health status of a systematic sample of 198 male and female inmates incarcerated in a large county jail located in a medium-sized Southern city. Using multiple regression analysis, predictors of physical health status, utilization of medical care, and inmates' evaluations of the accessibility and quality of health care are identified. The results indicate that gender and age are the most consistent demographic predictors of health status and medical care utilization, with females and older inmates reporting higher morbidity and concomitantly higher numbers of medical encounters. The experience of incarceration also appears to influence the physical health of inmates, as self reported health problems increase with inmates' duration of incarceration. Evaluations of jail medical care differ significantly by gender, with female inmates reporting more difficulty accessing health services, yet higher satisfaction with the quality of services received. The results suggest a need for medical care in correctional settings to adapt to the medical needs of older inmates and women, in addition to improving treatment for chronic conditions and preventive services. PMID- 10463473 TI - Incidental findings in a federally-sponsored cancer screening program. AB - The volume of non-cancer related clinical services and referrals for medical care of women as a consequence of their enrollment in a federally-sponsored breast and cervical cancer screening program was examined. We randomly sampled 100 medical records from among 389 individuals who received cancer screening services through the Connecticut Breast and Cervical Cancer Early Detection Program. Medical record audits tabulated occasions when women were offered or received diagnostic or therapeutic procedures as a by product of their program participation. Breast screening was provided to 100 women and 49 individuals received cervical cancer screening. In addition, 87 percent of the sample were offered or received one or more non-cancer related health services. Physical exams were provided to 86 women, laboratory tests were ordered for 11 individuals and 55 referrals were made to address a myriad of specific medical needs that were uncovered incidental to breast and cervical cancer screening. Among 26 women who did not heed recommendations for follow-up care, cost, inconvenience and beliefs that medical problems were not immediate concerns were cited. Local screening program sponsors should be cognizant that the health care needs and limited resources of some target populations may be substantial. Mechanisms to assure that needed health care is available to individuals should be built and into all categorical health service programs. PMID- 10463474 TI - Improved access to women's health services for Alaska natives through community health aide training. AB - This project demonstrates the effect of increasing the skills of Community Health Aides (CHAs) on the use of specific preventive health services by women in remote Alaska villages. Eight CHAs were trained in specimen collection for Pap and sexually transmitted disease testing, and in clinical breast examination. Skill competency was monitored. Computerized medical records of all women between the ages of 18 and 75 in the four villages with trained CHAs and in four comparison villages (n = 1093) were checked for Pap status prior to CHA training and again 12 months later. All eight CHAs achieved competency and provided services in their village clinics with telephone support from an experienced clinician. The post-training year Pap test rate of women who were overdue for a Pap test was 0.44 in the villages with trained CHAs; the rate among the women in the comparison villages was 0.32 (p = .079). PMID- 10463475 TI - Levels of expression of p27KIP1 protein in human prostate and prostate cancer: an immunohistochemical analysis. AB - p27KIP1 is a member of the CIP/KIP family of cyclin-dependent kinase inhibitory proteins that negatively regulate cell proliferation. Recent studies reported decreased p27 expression in breast and colon carcinomas and found that the loss of p27 is associated with a poor prognosis. We report here the results of our immunohistochemical analysis of p27 in human prostate cancer. Formalin-fixed, paraffin-embedded, whole-mount sections of prostate cancer from 73 selected patients treated by radical retropubic prostatectomy were obtained from the Department of Pathology, The Methodist Hospital, Houston, Texas. Ten histologically normal and nine high-grade prostatic intraepithelia neoplasia foci were selected from these whole-mount sections, and nine cases of transplant donor prostates were chosen as controls. Also, 10 prostate cancer metastatic lymph nodes were used to compare with the primary cancer group. Sections were immunostained with a monoclonal antibody against p27 protein using the avidin biotin complex immunohistochemical method. Immunoactivity was evaluated without knowledge of follow-up and recorded as the p27 labeling index (LI) (defined as the percentage of p27-positive cells among epithelia of the same category). The p27 (LI) in normal prostatic epithelia was 86.4+/-3.5% (the mean +/- the standard error of the mean). In contrast, the p27 immunoreactivity was significantly lower in cancers (LI: 43.5 +/-3.7%, P < .001) and in the high-grade prostatic intraepithelial neoplasia group (LI: 59.3 +/- 3.2%, P < .05). Expression of p27 in the metastatic lymph node group was significantly lower than in the other groups, including the prostate cancer cases and the cases of high-grade intraepithelial neoplasia (LI, 7.0%; P = .05). There was no association of the mean p27 LI with progression after radical prostatectomy. Nonrecurrent cases, with a mean follow-up time of greater than 5 years (n = 45), equalled 41.9%; recurrent cases, with a mean follow-up time of 18.3 months (n = 28), equalled 40.0%. The mean p27 LI was not associated with pathologic stage. Organ-confined specimens (n = 21) equalled 34.2%; cases of extraprostatic extension (n = 24) equalled 46.5%; and samples showing seminal vesicle involvement (n = 14) equalled 47.6%. In 14 cases with lymph node metastases, the mean p27 LI was 48.1% in the primary cancer (P = .2322). There was no association of the mean p27 LI with the Gleason score (P = .4747) nor with the clinical stage (P = .9914). PMID- 10463476 TI - Calponin and h-caldesmon in soft tissue tumors: consistent h-caldesmon immunoreactivity in gastrointestinal stromal tumors indicates traits of smooth muscle differentiation. AB - Currently, the immunohistochemical evaluation of smooth muscle differentiation is usually based on desmin, which also reacts with skeletal muscle and is not present in all smooth muscle tumors, and alpha-smooth muscle actin, which reacts with myoepithelial cells. Neither marker typically reacts with gastrointestinal stromal tumors (GISTs), previously classified as smooth muscle tumors or presently often classified as smooth muscle/stromal tumors. Two cytoskeleton associated actin-binding proteins, calponin (CALP) and h-caldesmon (HCD), are putative smooth muscle markers that also react with myoepithelia. These markers are of particular interest in the immunohistochemical analysis of tumors; neither of them has been extensively documented in soft tissue tumors. In this study, we evaluated selected normal and reactive tissues and more than 250 mesenchymal tumors for CALP and HCD. Both markers were expressed in parenchymal and vascular smooth muscle cells in various organs and in myoepithelial cells. CALP also reacted with myofibroblasts of desmoplastic stroma. All of our 25 benign smooth muscle tumors from various locations were positive for CALP and HCD, as were most of the retroperitoneal and uterine leiomyosarcomas. HCD was more specific, because CALP also reacted with myofibroblastic lesions. The common reactivity of malignant fibrous histiocytomas with CALP and HCD suggests a combination of myofibroblastic and smooth muscle differentiation in these tumors. The GISTs (c kit positive, usually actin negative) showed nearly consistent HCD reactivity, suggesting traits of smooth muscle differentiation. GISTs were usually CALP negative and showed a CALP expression pattern similar to that of alpha-smooth muscle actin. Although nonmuscle, nonmyofibroblastic tumors were negative for CALP and HCD, synovial sarcomas showed streaks of CALP-positive cells of unknown significance. CALP and HCD should be explored as markers to identify myofibroblastic and smooth muscle cell differentiation in mesenchymal tumors. PMID- 10463477 TI - Loss of heterozygosity in dysplasia and carcinoma of the gallbladder. AB - The loss or inactivation of genes at specific chromosomal loci is one of the important mechanisms during the tumor development in humans. To investigate the role of genetic alterations in the carcinogenesis of gallbladder carcinoma, 32 carcinoma cases and 11 dysplasia cases of gallbladder were analyzed for loss of heterozygosity (LOH) and microsatellite instability (MI) on chromosomal regions 3p, 5q, 8p, 9p, 13q, 17p, and 18q with 17 microsatellite markers. Loss of one allele was identified on chromosomes 5q (55%) and 17p (40%) in dysplasias and on chromosomes 3p (52%), 5q (66%), 9p (52%), and 17p (58%) in carcinomas. LOH on chromosomes 13q and 18q was frequent only in advanced stage (III and IV) carcinomas (40% and 31%, respectively). LOH on chromosome 17p was correlated with intranuclear p53 accumulation. LOH on multiple chromosomes was more frequent in advanced carcinomas with metastasis than in cases without metastasis (P < .05). A widespread MI was observed in only one case of carcinoma. We conclude that LOH on 5q is an early change of carcinogenesis in gallbladder and that LOH on 3p and 9p is related to the progression of gallbladder carcinoma LOH on 13q and 18q is likely to be a late event. LOH on 17p occurs not only in dysplasia but also increases during the subsequent stages. Accumulation of LOH may be associated with carcinogenesis of the gallbladder, but the role of MI may not be significant. PMID- 10463478 TI - Vascular endothelial growth factor expression in malignant melanoma: prognostic versus diagnostic usefulness. AB - Vascular endothelial growth factor (VEGF), an endothelial cell mitogen, plays a role in angiogenesis and progression in malignant melanoma. VEGF expression was examined in 62 biopsy specimens of melanocytic proliferations, including 45 malignant melanomas, 3 cellular blue nevi, 12 atypical compound nevi, and 2 Spitz nevi. The cases of malignant melanoma included 11 in situ melanomas, 18 Clark Level II, 9 Clark Level III, and 7 Clark Level IV tissue samples. All of the specimens were fixed in formalin and embedded in paraffin. Cytoplasmic immunoreactivity for VEGF was demonstrated in 19 (42%) of 45 melanoma samples, but there was no immunoreactivity for VEGF exhibited by any of the atypical compound melanocytic nevi, cellular blue nevi, or Spitz nevi (P < .009). Immunoreactivity for VEGF was found to be related to tumor thickness (as evidenced by Clark level [P < .03]) and to absence of regression (P < .04). Although VEGF is not a useful prognostic indicator for malignant melanoma, it may be useful as a discriminating factor between malignant melanoma and benign melanocytic lesions, and it may offer some insight into tumor growth. PMID- 10463479 TI - H-ras and K-ras gene mutations in primary human soft tissue sarcoma: concomitant mutations of the ras genes. AB - ras gene mutations have been described with varying frequency in several types of human malignancies. To determine the incidence and type of ras mutations in human soft tissue tumors, we studied 45 sarcomas, including 27 malignant fibrous histiocytomas (MFHs), 10 liposarcomas, 2 rhabdomyosarcomas, and 6 leiomyosarcomas. Al of the tumors were investigated by direct sequence analysis with the automated DNA sequencing of polymerase chain reaction-amplified ras sequences. Twenty (44%) of the sarcomas examined harbored K-ras mutations, 18 (90%) of which were MFHs. All of the K-ras mutations were G-to-A transition mutations in the second position of codon 13 (glycine --> aspartic acid). Of the samples with K-ras activation, 7 (16% of the total of 45 tumors), including 6 MFHs and 1 leiomyosarcoma, also contained H-ras mutation. All of the tumors that showed H-ras alteration had G-to-T transversion mutations in the second base of codon 12 (glycine --> valine). These data possibly implicate that ras gene activation may be a relatively uncommon event in soft tissue tumors, with the exception of MFH. It is suggested that the oncogenic process underlying the development of tumors between these groups may be different and that ras gene mutations may play a role in the etiology and/or progression of MFH. It is noteworthy that when ras gene activation occurs in sarcoma, it predominantly affects the K-ras gene, particularly codon 13. Moreover, H-ras mutations in our samples were detected only in association with tumors that also displayed K-ras mutated genes. This study demonstrates for the first time concomitant mutations in two different members of the ras gene family in sarcoma PMID- 10463480 TI - Submission of lymph node tissue for ancillary studies decreases the accuracy of conventional breast cancer axillary node staging. AB - Pathologists are under increasing pressure to submit fresh tissue for ancillary studies and research protocols. In several tumor types (breast, lung, melanoma, colorectal, prostate), increased interest in detecting submicroscopic nodal metastases through reverse transcriptase polymerase chain reaction analysis of mRNA from portions of lymph nodes has precluded histologic analysis of the entire node for metastases. A retrospective review was undertaken of 227 breast cancer patients prospectively entered on a research protocol examining the usefulness of sentinel lymph node surgery. All of the patients ultimately underwent complete lymph node dissection. The research protocol required that all nodes greater than 8 mm in size be bisected and submitted separately. Positive lymph nodes were evaluated for unilateral or bilateral involvement in the node sections. Sixty node-positive patients were identified, yielding 230 positive nodes. One hundred seven of these nodes were confirmed to have been bisected. Carcinoma was identified in both lymph node sections in 64 (59.8%) nodes and in only one-half of the bisected lymph node in 43 (40.2%) nodes. Involvement of both sections was more likely when patients had multiple nodes positive. In 12 patients, involvement of one-half of the bisected nodes was the only evidence of metastatic disease (20.0% of node-positive patients). This evidence suggests that submission of less than the complete lymph node for histologic evaluation of metastatic disease decreases the accuracy of lymph node staging. Furthermore, a significant proportion of patients may be erroneously classified as histologically node negative. PMID- 10463481 TI - Epidermal growth factor, estrogen, and progesterone receptor expression in primary sweat gland carcinomas and primary and metastatic mammary carcinomas. AB - The distinction between primary sweat gland carcinomas and metastatic breast carcinoma to the skin is sometimes difficult. In an effort to improve this discrimination, we compared the immunohistochemical staining pattern of 42 primary sweat gland carcinomas (SGCs) with 30 metastases from breast carcinoma (BC) to the skin, 125 primary BCs, and 30 noncutaneous metastases from BCs. The antibodies used were against the receptors for epidermal growth factor (EGF-R), estrogen receptor (ER), and progesterone receptor (PR). The frequencies of positive staining were as follows for EGF-R: 34 (81%) of 42 SGCs, 5 (17%) of 30 BCs metastatic to skin, 28 (22%) of 125 primary BCs, and 6 (20%) of 30 noncutaneous BC metastases. For ER, the frequencies were 9 (21%) of 42 SGCs and 10 (33%) of 30 BCs metastatic to skin. The frequencies for PR were 8 (19%) of 42 SGCs and 8 (27%) of 30 BCs metastatic to skin. These results suggest that expression of EGF-R may be diagnostically helpful, because it is strongly associated with SGCs when compared with metastatic BCs (P < .0001). This association is also present when ductal eccrine and apocrine types of SGC, which are the histologic subtypes of SGC most difficult to distinguish from metastatic BC, are separately analyzed (P < .001). The frequencies of expression of ER and PR in SGCs and BCs metastatic to skin were not significantly different. PMID- 10463482 TI - Comparison of different polymerase chain reaction-based approaches for clonality assessment of immunoglobulin heavy-chain gene rearrangements in B-cell neoplasia. AB - Several frequently applied polymerase chain reaction strategies for analysis of immunoglobulin heavy-chain gene rearrangements were compared by analyzing 70 B cell lymphoproliferative disorders and 24 reactive lymphoid lesions. Southern blot analysis was used as the "gold standard" for clonality assessment. For polymerase chain reaction analysis, primers directed against framework (FR) 3 (FR3-A and FR3-B), FR2, and FR1 of the variable gene segments and against joining gene segments of the immunoglobulin heavy-chain gene were used. Polymerase chain reaction products were analyzed by high-resolution fingerprinting analysis using radiolabeled nucleotides, gene scanning using fluorochrome-labeled primers, and heteroduplex analysis. All of the assays generated polyclonal patterns in the reactive tissues. The sensitivity in detecting monoclonality as defined by Southern blotting varied between 60% (heteroduplex analysis with FR3 primers) and 77% (high-resolution fingerprinting analysis with FR2 primers). Comparison of the three FR3 assays revealed that gene scanning had the highest sensitivity (69%), probably because it could detect small aberrant monoclonal amplicons. False negative results were especially frequent in follicular center lymphoma (n = 20), but also in diffuse large B-cell lymphoma (n = 18), both renowned for having mutated variable gene segments, potentially leading to primer mismatching. For example, in follicular center lymphoma, the FR3, FR2, and FR1 region primer sets detected clonality in only 35 to 40, 65, and 50%, respectively. Combining these techniques, 17 (85%) of 20 follicular center lymphoma samples showed monoclonality. Therefore, especially in follicular center lymphoma, diffuse large B-cell lymphoma, and, to a lesser extent, in other lymphomas, multiple variable gene segment primer sets must be used for a reliable assessment of clonality. Our results also suggest that gene scanning is somewhat more sensitive than other read-out systems. Heteroduplex analysis, however, is a reliable alternative within a diagnostic setting, avoiding the use of radioactivity or expensive automated sequencing equipment and fluorochrome-labeled (oligo)nucleotides. PMID- 10463483 TI - Quantitative assessment of bladder cancer by nuclear texture analysis using automated high resolution image cytometry. AB - The aim of this study was to investigate the possibility of identifying urothelial neoplasia by nuclear chromatin texture feature analysis using high resolution image cytometry to improve the diagnostic accuracy of cytologic examination in the detection and monitoring of bladder cancer. Touch imprints of transurethral resection material of 56 control group (CG) cases of nonmalignant urothelium and 94 tumor group (TG) cases of bladder cancer were analyzed. The specimen collection was divided randomly into a training set and a test set. Cells were stained specifically for DNA by the Feulgen method. Only diploid cell nuclei were analyzed from both groups. A discriminator comprised of three nuclear texture features was derived from the training set of cases to separate CG from TG cases. This discriminator was then applied to the independent test set. CG cases were separated from TG cases with a sensitivity of 97% and a specificity of 95% on the independent test set of cases. When dividing TG cases into high-risk and low-risk groups, sensitivity in the low-risk group was 93%. None of the high risk cases was misclassified (sensitivity, 100%). This retrospective investigation demonstrates that by high-resolution image cytometry it is possible to distinguish between urothelial neoplasia and normal urothelium with high reliability when examining diploid cell nuclei only. This method is superior to DNA ploidy analysis using image or flow cytometry and may become clinically relevant as a supplement to conventional cytologic examination. These promising results should be confirmed on cytologic preparations derived from bladder washings or voided urine. PMID- 10463484 TI - Paraneoplastic limbic encephalitis associated with small cell carcinoma of the prostate. AB - A 76-year-old man with primary small cell carcinoma of the prostate died after a subacute illness marked by memory loss and truncal ataxia Post-mortem examination of the central nervous system was consistent with limbic encephalitis and cerebellar degeneration. Although limbic encephalitis is a known complication of small cell carcinoma of the lung, this seems to be the first reported case of limbic encephalitis associated with small cell carcinoma of the prostate. Implications with respect to diagnosis and therapy are discussed. PMID- 10463485 TI - Oligoclonal immunoglobulin heavy-chain and T-cell receptor delta rearrangements persist in a recurrent acute lymphoblastic leukemia with one immunoglobulin kappa rearrangement as a clonal marker. AB - Acute lymphoblastic leukemias (ALLs) represent the clonal expansion of a lymphoid precursor cell. Therefore, all cells of an ALL should have identical antigen receptor gene rearrangements. In a patient with diploid ALL of the B-cell precursor immunophenotype, seven different clonal rearrangements of the immunoglobulin heavy-chain genes (IgH) were identified, implying the presence of oligoclonal populations. All of these rearrangements were only detectable after a modification of the polymerase chain reaction for the complementarity determining region of the IgH genes using V(H) gene framework 3 and (H) consensus primers. Sequence analysis showed that these rearrangements were completely unrelated to each other. Only two of these rearrangements were detectable by Southern blot analysis. Quantification and single-cell analysis confirmed the high frequency of these latter two rearrangements, as well as their presence in the same clonal population. The other rearrangements characterized less than 5% of the leukemic population. In addition, two T-cell receptor Vdelta2-Ddelta3 (TCRdelta) rearrangements were identified, both at a similar frequency. However, they were derived from different cells. An Igkappa rearrangement represented the only clonal marker in this leukemia. All of the Ig and TCRdelta rearrangements, with the exception of one IgH rearrangement, remained stable throughout the course of the disease. The persistence of such a great number of distinct IgH rearrangements at different quantities within the leukemic population and of the two biclonal TCRdelta rearrangements is compatible with the presence of a clonal disease that is defined by the Igkappa rearrangement. PMID- 10463486 TI - Evaluation of HER-2/neu (erbB-2) status in breast cancer: from bench to bedside. AB - Molecular alterations in breast cancer are being incorporated into the development of new treatment strategies. The HER-2/neu oncogene has been extensively investigated as a prognostic factor and recently as a predictor of response to chemotherapy or endocrine therapy. The development of a humanized anti-HER-2 monoclonal antibody (Herceptin) and the encouraging results obtained in the treatment of patients with HER-2 overexpressing metastatic breast cancer with this antibody have resulted in renewed interest in HER-2/neu. This article reviews the current knowledge of HER-2/neu both as a prognostic and a predictive factor. Problems associated with the standardization of the methodology for assessing HER-2/neu status and clinically significant cut-off points are addressed. PMID- 10463487 TI - Recommendations for reporting of tumors of the adrenal cortex and medulla. Association of Directors of Anatomic and Surgical Pathology. PMID- 10463488 TI - Correspondence re: Crescenzi A, Panunzi C, Taccogna S, Papini E, Pacella CM, NARDI F: cytospray fixation of frozen intraoperative sections. Mod pathol 12:92 94, 1999. PMID- 10463489 TI - Correspondence re: Jatoi A, Dallal GE, Nguyen PL: false-negative rates of tumor metastases in the histologic examination of bone marrow. Mod pathol 1999;12:29 32. PMID- 10463490 TI - Rotary blood pumps: a new horizon. PMID- 10463491 TI - Flow visualization study to improve hemocompatibility of a centrifugal blood pump. AB - A correlation study was conducted among quantitative flow visualization analysis, computational fluid dynamic analysis, and hemolysis tests regarding the flow in a centrifugal blood pump to prevent hemolysis. Particular attention was paid to the effect of the impeller/casing gap widths on the flow in the volute and in the outlet. Flow vector maps were obtained for 250% scaled-up models with various geometries, using an argon ion laser light sheet, a high speed video camera, and particle tracking velocimetry. In terms of the results, in the small radial gap model, high shear occurred near the inside wall of the outlet and stagnation near the outside wall of the outlet whereas the standard model maintained smooth flow and low shear. The small radial gap model showed a lower head and greater hemolysis than the standard model. This head decrease could be partly restored by relocating the outlet position; however, the hemolysis level hardly decreased. From these results, it was found that the small radial gap itself is important. It was also confirmed by detailed flow visualization and simple laminar shear analysis near the wall that the small radial gap caused a wider high shear layer (110-120 microm) than the standard model (approximately 80 microm). In the small radial gap model, the high shear layer in the outlet (approximately 50 microm) is much narrower than that in the volute. Flow visualization together with the aid of computational fluid dynamic analysis would be useful to eliminate the causes of hemolysis. PMID- 10463492 TI - An emergency balloon occlusion system for a rotary blood pump left ventricular assist system. AB - A fatal outcome is expected in a left ventricular assist system (LVAS) utilizing a rotary blood pump if there is no mechanism to prevent the backflow from the aorta to the heart in the case of acute pump failure. To solve this problem, a passive mechanical clamping system at the outflow graft of a rotary blood pump was developed together with Fuji Systems, Inc., Yokohama, Japan. The system consisted of an emergency clamp port and an occlusion balloon. The balloon was fixed around the outlet graft of the LVAS. In an in vitro study, a fail-safe clamping operation with 2 ml saline injection under 7 L/min flow against 140 mm Hg pressure reduced the flow to 0.5 L/min while the pressure in the system increased to 190 mm Hg. The systems were also applied to 2 in vivo LVAD studies. When the pumps were stopped, there were approximately 3.0 L/min regurgitant flows. The balloon occluder prevented this regurgitant flow effectively against a 100/80 mm Hg arterial pressure. In conclusion, this emergency balloon occlusion system is relatively easy to operate and will work efficiently in all possible clinically encountered malfunctions of the rotary blood pump LVAS. PMID- 10463493 TI - Sealing properties of mechanical seals for an axial flow blood pump. AB - A miniature intraventricular axial flow blood pump for left ventricular support is under development. One of the key technologies required for such pumps is sealing of the motor shaft. In this study, to prevent blood backflow into the motor side, mechanical seals were developed and their sealing properties investigated. In the experimental apparatus, the mechanical seal separated the bovine blood on the chamber side from the cooling water on the motor side. A leakage of the blood was measured by inductively coupled plasma (ICP) light emission analysis. The rate of hemolysis was measured by the cyanmethemoglobin method. Frictional torque acting on the shaft was measured by a torque transducer. In the experiments, the rotational speed of the shaft was changed from 1,000 to 10,000 rpm, and the contact force of the seal faces was changed from 1.96 to 4.31 N. To estimate lubrication regimes, the Stribeck curve, a diagram of the coefficient of friction against the bearing characteristic G number, was drawn. The results of the experiments showed that both the leakage of blood and the rate of hemolysis were very small. The friction loss was also very small. The mechanical seal was operated in various lubrication regimes, from a fluid lubrication regime to a mixed lubrication regime. PMID- 10463494 TI - Defining pulsatile perfusion: quantification in terms of energy equivalent pressure. AB - Several clinical and animal studies have demonstrated that pulsatile perfusion is more beneficial than nonpulsatile perfusion during short or long durations of extracorporeal circulation. Other investigators, however, have been unable to document these benefits. The issue remains controversial. Central to the debate is the issue of a precise definition of pulsatile flow. To help resolve the conflict, pulsatile flow may be quantified in terms of energy equivalent pressure. This formula contains both the arterial pressure and pump flow rate, which are the 2 most critical parameters for open heart surgery. This definition establishes common criteria for assessment of the effectiveness of extracorporeal support. PMID- 10463495 TI - The effects of pulsatile versus nonpulsatile perfusion on blood viscoelasticity before and after deep hypothermic circulatory arrest in a neonatal piglet model. AB - Blood trauma increases blood viscoelasticity by increasing red cell aggregation and plasma viscosity and by decreasing cell deformability. During extracorporeal circulation, the mode of perfusion (pulsatile or nonpulsatile) may have a significant impact on blood trauma. In this study, a hydraulically driven dual chamber pulsatile pump system was compared to a standard nonpulsatile roller pump in terms of changes in the blood viscosity and elasticity during cardiopulmonary bypass (CPB) and pre and post deep hypothermic circulatory arrest (DHCA). Piglets, with an average weight of 3 kg, were used in the pulsatile (n = 5) or nonpulsatile group (n = 5). All animals were subjected to 25 min of hypothermia, 60 min of DHCA, 10 min of cold reperfusion, and 40 min of rewarming with a pump flow of 150 ml/kg/min. A pump rate of 150 bpm, pump ejection time of 120 ms, and stroke volume of 1 ml/kg were used during pulsatile CPB. Arterial blood samples were taken pre-CPB (36 degrees C), during normothermic CPB (35 degrees C), during hypothermic CPB (25 degrees C), pre-DHCA (18 degrees C), post-DHCA (19 degrees C), post-rewarming (35 degrees C), and post-CPB (36 degrees C). Viscosity and elasticity were measured at 2 Hz and 22 degrees C and at strains of 0.2, 1, and 5 using the Vilastic-3 Viscoelasticity Analyzer. Results suggest that the dual chamber neonate-infant pulsatile pump system produces less blood trauma than the standard nonpulsatile roller pump as indicated by lower values of both viscosity and elasticity during CPB support. PMID- 10463496 TI - Diminished vasoconstrictive function caused by long-term nonpulsatile left heart bypass. AB - We investigated the functional changes of the systemic vascular system due to prolonged nonpulsatile left heart bypass (NPLHB). Three adult goats underwent pulsatile left heart bypass (PLHB). Two weeks later the PLHB was changed to the NPLHB, which was conducted for 4 weeks. The aortic pulse pressure was 39 and 16 mm Hg during the PLHB and NPLHB, respectively. Systemic vascular resistance (SVR) and the plasma norepinephrine level were measured at the end of PLHB (PUL), and in the 1st, 2nd, 3rd, and 4th weeks of NPLHB (NP1w, NP2w, NP3w, and NP4w). At each point, 1 microg/kg norepinephrine was injected, and the elevation of the SVR (deltaSVR) was calculated. The SVR and the plasma norepinephrine level did not change significantly during the entire course. However, deltaSVR decreased during NPLHB and became significantly lower at NP3w and NP4w than that at PUL (NP3w: 839 +/- 164, NP4w: 746 +/- 268, and PUL: 1,239 +/- 324 dyne x s x cm(-5)). These results strongly indicated that prolonged NPLHB significantly diminished the constrictive function of the vascular system. PMID- 10463497 TI - Augmentative effect of pulsatility on the wall shear stress in tube flow. AB - Wall shear stress (WSS) has been considered to play an important role in the physiological and metabolic functions of the vascular endothelial cells. We investigated the effects of the pulse rate and the maximum flow rate on the WSS to clarify the influence of pulsatility. Water was perfused in a 1/2 inch transparent straight cylinder with a nonpulsatile centrifugal pump and a pulsatile pneumatic ventricular assist device (VAD). In nonpulsatile flow (NF), the flow rate was changed 1 to 6 L/min by 1 L/min increments to obtain standard values of WSS at each flow rate. In pulsatile flow (PF), the pulse rate was controlled at 40, 60, and 80 bpm, and the maximum flow rate was varied from 3.3 to 12.0 L/min while the mean flow rate was kept at 3 L/min. The WSS was estimated from the velocity profile at measuring points using the laser illuminated fluorescence method. In NF, the WSS was 12.0 dyne/cm2 at 3 L/min and 33.0 dyne/cm2 at 6 L/min. In PF, the pulse rate change with the same mean, and the maximum flow rate did not affect WSS. On the other hand, the increase in the maximum flow rate at the constant mean flow rate of 3 L/min augmented the mean WSS from 13.1 to 32.9 dyne/cm2. We concluded that the maximum flow rate exerted a substantial augmentative effect on WSS, and the maximum flow rate was a dominant factor of pulsatility in this effect. PMID- 10463498 TI - Total vascular resistance and blood flow frequency during left ventricular assistance using a vibrating flow pump. AB - A vibrating flow pump (VFP) can generate high frequency oscillated blood flow within 10-30 Hz by the oscillation of its central tube. A totally implantable artificial heart using a VFP is being developed as a unique type of blood pump. In this study, left ventricular (LV) assist circulation was performed using a VFP. The total vascular resistance and driving frequency of the VFP were estimated from their relationship. The effect of oscillation on the vascular system was studied by the frequency analysis method and vascular impedance. Adult goats were anesthetized by halothane using an inhaler and a left fourth thoracotomy was performed. The inflow cannula was inserted into the left ventricle, and the outflow cannula was sutured to the descending aorta. The VFP and a centrifugal pump were set in parallel for alternation and comparison. The driving frequency of the VFP was changed and included 15, 20, 25, and 30 Hz. The hemodynamic parameters were continuously recorded during experiments by a digital audio tape (DAT) data recorder. The internal pressure of the left ventricular cavity and aortic pressure were monitored by the pressure manometers continuously. One hundred percent LV assistance was judged by the separation of LV and aortic pressure. The total vascular resistance was decreased by the start of operation of each pump. The decrease during flow using the VFP was not as large as that using a centrifugal pump (CP). The arterial input impedance during oscillated blood flow by the VFP showed a slow curve appearance. It was similar to the frequency characteristics curve of natural heart beats within the lower frequencies. The study of arterial impedance may be important for the estimation of the reflection of the pulsatile wave from the arterial branch, among other things. PMID- 10463499 TI - Recovery from lymphocytopenia following extracorporeal circulation: simple indicator to assess surgical stress. AB - This study investigated whether the lymphocyte count is a useful indicator to assess surgical damage following extracorporeal bypass. In Study 1, to investigate the correlation between extracorporeal circulating time (ECCT) and lymphocyte counts, 40 elective CABG patients were studied retrospectively. The lymphocyte recovery ratio (LRR), which represented the actual lymphocyte count divided by the preoperative lymphocyte count, was determined preoperatively, and on postoperative day (POD) 1, POD 3, and POD 5. In Study 2, the correlation between the interleukin-8 (IL-8) level and LRR was examined prospectively in elective CABG patients (n = 20). We measured the LRR and serum IL-8 levels preoperatively and during extracorporeal circulation (ECC) at 5 min, at the end of ECC, and 1, 3, and 12 h following ECC termination. Study 1 showed that the LRR decreased until POD 1 and gradually increased thereafter. The LRR had a negative correlation with the ECCT. In Study 2, the IL-8 level demonstrated a time course opposite to that of the LRR; it increased until 3 h after ECC termination and declined thereafter. There was a significant negative correlation between the LRR on POD 3 and the IL-8 level at 3 h after ECC termination. In summary, long-term ECC induced significant and prolonged lymphocytopenia. The LRR had a negative correlation with IL-8. These results indicated that the LRR may represent the degree of surgical stress following ECC; therefore, the counting of lymphocytes can be a quite useful bedside monitor to assess surgical damage and prognosis. PMID- 10463500 TI - Flow balance between the left and right cardiac output of an eccentric roller type total artificial heart. AB - We have made an eccentric roller type total artificial heart (ERTAH). The ERTAH is a positive-displacement device comparable to a DeBakey roller pump. Its left and right outputs are determined by the size of its blood chambers, and the ratio of its left and right output is almost constant. We focused on an interatrial shunt to achieve left-right balance. We have conducted numerical simulation, a mock test, and an acute animal experiment to analyze left-right heart balance during ERTAH operation. Numerical simulation was performed under conditions in which the flow of the left artificial heart was fixed at 6 L/min, the flow of the right artificial heart was varied from 4.8 to 6 L/min, and the interatrial resistance was also varied. The relationship between the interatrial shunt flow rate and the output of the left and right artificial hearts was balanced when the flow of the right artificial heart was at 5.45 L/min. In a mock test, 2 DeBakey roller pumps were connected to the left and right sides of a Donovan mock circulatory system, and an interatrial shunt was created between the inlet ports of the left and right roller pumps. The interatrial resistance of the mock system was varied from 7.7, to 4.3, and to 2.9 mm Hg x min/L when the inner diameter of the interatrial shunt was 6, 8, and 10 mm, respectively. As in the mock test, 2 roller pumps were used to bypass the right and left hearts of a goat weighing 60 kg. The flow rate of the left heart was almost constant (4.7 L/min). The flow of the right heart was approximately 4.1 L/min when the interatrial shunt flow rate was zero. A leading consideration was that the left to left shunt through the bronchial arteries in this goat was approximately 0.6 L/min. In developing the ERTAH, we considered that creating an interatrial shunt between the inlet ports of the ERTAH as well as making a difference between the chamber volumes might be effective in balancing the left-right sides of the artificial heart. PMID- 10463501 TI - Peripheral vascular resistances during total left heart bypass with an oscillated blood flow. AB - For development aimed at a totally implantable type ventricular assist device (VAD), the vibrating flow pump (VFP) has been developed at Tohoku University. A transcutaneous energy transmission system (TETS) using amorphous fibers was developed to power the totally implantable VAD system. The VFP works at a high frequency compared to that of a natural heart of a biological system. It is a frequency of 10-50 Hz. In this research, animal experiments with left heart bypass were carried out with healthy adult goats. For comparison between nonpulsatile flow and oscillated flow, a rotary pump (RP) and the VFP were used in the experiments. For the achievement of total left heart bypass, left ventricular approaches were carried out, and blood was pumped from the left ventricle to the descending aorta. Adequate support of the left heart was provided by both pumps. In terms of the results, the vascular resistances tended to decrease during the use of both pumps during 100% bypass driving. When we compared these pumps at the same flow rate, the resistances during RP driving were significantly smaller than those during VFP driving. These results may suggest that the influences of the VFP upon the peripheral vessels may be relatively small compared to those of the RP. This may be an important result when a stable hemodynamic condition is required during artificial circulation. The VFP was considered as a candidate for a totally implantable VAD as a result. PMID- 10463502 TI - Detection of the cardiac function by fractal dimension analysis. AB - Nonlinearity in circulation control attracts attention because nonlinearity is thought to be essential in the function of the living body. Many investigators have pointed out that the analysis of heart rate variability in particular is important in the analysis of autonomic nerve and cardiac function evaluation. Heart rate variability shows nonlinear behavior. However, until the present, many reports have been premised on linearity; linear correlation by frequency analysis has been used by many studies. However, in terms of this methodology, there is a problem applying it to the nonlinear living body. Therefore, fractal and chaos methodology has been used. The ascertainment of cardiac function has become important in allowing the clinical stage of a ventricular assist system to be successful. The purpose of this study was cardiac function evaluation by a methodology that was premised on nonlinearity. Chaos and fractal theory was used as a nonlinear dynamic theory. As a methodology of measurement, the volume of the left ventricle was used rather than an electrocardiogram, the waveform of arterial blood pressure. The volume was measured using acoustic quantification (AQ) ultrasonic echocardiography. Using these methodologies, the time series of many patients were analyzed. For example, drug administration was attempted in this study, and it was found that some drugs like ACE inhibitors showed a significant effect upon nonlinear dynamics in the cardiovascular system. The result, which attempted cardiac function evaluation by these various methodologies, is reported. PMID- 10463503 TI - Development of design methods for a centrifugal blood pump with a fluid dynamic approach: results in hemolysis tests. AB - The purpose of this study was to examine the relationship between local flow conditions and the hemolysis level by integrating hemolysis tests, flow visualization, and computational fluid dynamics to establish practical design criteria for centrifugal blood pumps with lower levels of hemolysis. The Nikkiso centrifugal blood pump was used as a standard model, and pumps with different values of 3 geometrical parameters were tested. The studied parameters were the radial gap between the outer edge of the impeller vane and the casing wall, the position of the outlet port, and the discharge angle of the impeller vane. The effect of a narrow radial gap on hemolysis was consistent with no evidence that the outlet port position or the vane discharge angle affected blood trauma in so far as the Nikkiso centrifugal blood pump was concerned. The radial gap should be considered as a design parameter of a centrifugal blood pump to reduce blood trauma. PMID- 10463504 TI - Computational fluid dynamics analysis to establish the design process of a centrifugal blood pump: second report. AB - To establish an efficient design process for centrifugal blood pumps, the results of computational fluid dynamics (CFD) analysis were compared to the results of flow visualization tests and hemolysis tests, using the Nikkiso centrifugal blood pump. CFD analysis revealed that the radial gap greatly affected the shear stress in the outlet diffuser. The hemolysis study also indicated a similar tendency. To see the flow behind the impeller, we conducted a comparative study between models with and without washout holes using the CFD technique. CFD analysis indicated that flow and pressure distributions behind the impeller were different between both models, and a particle was observed to remain longer behind the impeller in the model without washout holes. In the future, CFD analysis could be a useful tool for developing blood pumps in comparison to flow visualization tests and hemolysis tests. PMID- 10463505 TI - Blood flow in a continuous flow ventricular assist device. AB - A numerical analysis was performed to predict the shear stresses, flow rates, and the velocity profiles in a continuous flow ventricular assist device, the CFVAD3. The problem was modeled as a rotating disk over a stationary disk. A variety of clearances was tested for the CFVAD3 coupled with a range of rotational speeds and pressure gradients. Velocity fields were generated using solutions obtained with FLOW3D software (AEA Technology, Pittsburgh, PA, U.S.A.) Analysis of these solutions shows that the pressure differential effect has a stronger influence on the flow than the rotational effect of the impeller Ekman layer. The predicted shear stresses reflect these changes in the volume flow rates and the speeds shown in the velocity profiles. Based on the predictions of the software, the optimum clearance and rotational speed were chosen. The conclusion is that a speed in the range of 2,200-2,400 rpm should be chosen depending on the efficiency of the pump. PMID- 10463506 TI - Feasibility of a tiny Gyro centrifugal pump as an implantable ventricular assist device. AB - The Gyro pumps were developed for long-term circulatory support. The first generation Gyro pump (C1E3) achieved 1 month paracorporeal circulatory support in chronic animal experiments; the second generation (PI702) implantable ventricular assist device (VAD) was successful for over 6 months. The objective of the next generation Gyro pump is for use as a long-term totally implantable VAD and for pediatric circulatory support. This tiny Gyro pump (KP101) was fabricated with the same design concept as the other Gyro pumps. The possibility of an implantable VAD was determined after performance and hemolysis test results were compared to those of the other Gyro pumps. The pump housing and impeller were fabricated from polycarbonate with an impeller diameter of 35 mm. The diameter and height of the pump housings are 52.3 mm and 29.9 mm, respectively. At this time, a DC brushless motor drives the KP101, which is the same as that for the C1E3. The pump performance was measured in 37% glycerin water at 37 degrees C. Hemolysis tests were performed utilizing a compact mock loop filled with fresh bovine blood in a left ventricular assist device (LVAD) condition at 37 degrees C. The KP101 achieved the LVAD conditions of 5 L/min and 100 mm Hg at 2,900 rpm; generated 10 L/min against 100 mm Hg at 3,200 rpm; 3 L/min against 90 mm Hg at 2,600 rpm; and 2 L/min against 80 mm Hg at 2,400 rpm. In addition, the pump efficiency during this experiment was 12.5%. The other Gyro pumps. that is, the C1E3, PI601, and PI701, in an LVAD condition require 1,600, 2,000, and 2,000 rpm, respectively. The KP101 produced a normalized index of hemolysis (NIH) value of 0.005 g/100 L. With regard to the NIH, the other Gyro pumps, namely the C1E3, PI601, and PI701 demonstrated 0.0007, 0.0028, and 0.004 g/100 L, respectively. The KP101 produced an acceptable pressure flow curve for a VAD. The NIH value was higher than that of other Gyro pumps, but is in an acceptable range. PMID- 10463507 TI - A sealless centrifugal blood pump with passive magnetic and hydrodynamic bearings. AB - We are developing a permanently implantable ventricular assist system based on a sealless centrifugal blood pump. The impeller of the pump is supported by a passive radial magnetic bearing acting in synergy with hydrodynamic bearings. Torque is transmitted to the impeller by electromagnetic coupling via an integrated axial flux gap motor. Computer modeling has been used extensively to guide the hydraulic and electromagnetic design of the pump. As part of the development effort, a prototype system was built, which consisted of a radial magnetic bearing, an axial air gap motor, and a pivot bearing to constrain the axial motion. The following testing has been completed to validate the design. First, hydraulic tests have demonstrated sufficient hydraulic performance. Second, preliminary in vitro evaluation of hemolysis was low compared to that of a BioPump control. Third, a 6 h in vivo experiment was successfully completed. PMID- 10463508 TI - Test controller design, implementation, and performance for a magnetic suspension continuous flow ventricular assist device. AB - A new continuous flow ventricular assist device using full magnetic suspension has been designed, constructed, and tested. The magnetic suspension centers the centrifugal pump impeller within the clearance passages in the pump, thus avoiding any form of contact. The noncontact operation is designed to give very high expected mechanical reliability, large clearances, low hemolysis, and a relatively small size compared to current pulsatile devices. A unique configuration of magnetic actuators on the inlet side and exit sides of the impeller provides full 5 axis control and suspension of the impeller. The bearing system is divided into segments which allow for 3 displacement axes and 2 angular control axes. The controller chosen for the first suspension tests consists of a decentralized set of 5 proportional integral derivative (PID) controllers. This document describes both the controller and an overview of some results pertaining to the magnetic bearing performance. The pump has been successfully operated in both water and blood under design conditions suitable for use as a ventricular assist device. PMID- 10463509 TI - Characterization of a magnetic bearing system and fluid properties for a continuous flow ventricular assist device. AB - This article presents the performance test results of the CFVAD3 continuous flow blood pump in an artificial human circulation system. The CFVAD3 utilizes magnetic bearings that support a thin pancake impeller, the shape of which allows for a very compact pump whose total axial length is less than 5 cm with a radial length of about 10 cm. This gives a total volume of about 275 cc. The impeller itself has 4 vanes with a designed operating point of 6 L/min at 100 mm Hg of differential pressure and 2,000 rpm. The advantages of magnetic bearings, such as large clearance spaces and no mechanical wear, are elaborated upon. Furthermore, bearing model parameters such as load capacity and current gains are described. These parameters in conjunction with the operating conditions during testing are then used to estimate the fluid forces, stiffness, and damping properties while pumping. Knowledge of these parameters is desirable because of their effects on pump behavior. In addition, a better plant model will allow more robust control algorithms to be devised that can boost pump performance and reliability. PMID- 10463510 TI - Rotary blood pump flow spontaneously increases during exercise under constant pump speed: results of a chronic study. AB - Many types of rotary blood pumps and pump control methods have recently been developed with the goal of clinical use. From experiments, we know that pump flow spontaneously increases during exercise without changing pump control parameters. The purpose of this study was to determine the hemodynamics associated with the long-term observation of calves implanted with centrifugal blood pumps (EVAHEART, Sun Medical Technology Research Corporation, Nagano, Japan). Two healthy female Jersey calves were implanted with devices in the left thoracic cavity. A total of 22 treadmill exercise tests were performed after the 50th postoperative day. During exercise, the following parameters were compared with conditions at rest: heart rate, blood pressure, central venous oxygen saturation (SvO2), pump speed, and pump flow. The pump flow in a cardiac cycle was analyzed by separating the systole and diastole. Compared to the base data, statistically significant differences were found in the following interrelated parameters: the heart rate (66.8 +/- 5.2 vs. 106 +/- 9.7 bpm), mean pump flow (4.8 +/- 0.2 vs. 7.0 +/- 0.3 L/min), and volume of pump flow in diastole (26.0 +/- 1.8 vs. 13.5 +/- 2.5 ml). During exercise, the volume of pump flow in systole was 3 times larger than that measured in diastole. Blood pressure, SvO2, and pump speed did not change significantly from rest to exercise. These results suggested that the mean pump flow depends on the systolic pump flow. Therefore, the increase in the mean pump flow during exercise under constant pump speed was caused by an increase in the heart rate. PMID- 10463511 TI - Chronic survival of calves implanted with the DeBakey ventricular assist device. AB - The DeBakey ventricular assist device (VAD) is a miniaturized, electromagnetically driven axial flow pump capable of generating in excess of 10 L/min output. The VAD was evaluated in 19 calves during experiments designed to test iterative modifications in the system and to determine the safety of the DeBakey VAD for intermediate to long-term implant. Five of the animals died or were euthanized during the perioperative period (i.e., Days 1-5) due to complications associated with bleeding (n = 3), sudden cardiac arrest (n = 1), or pump occlusion due to a muscle remnant associated with coring (n = 1). The remaining 14 animals survived from 7-145 days. Ten of the 14 animals survived 30 or more days, and 2 animals survived 93 and 145 days before elective euthanasia. Pump function was evaluated in the 14 calves that survived beyond the perioperative period. Pump output at implantation averaged 3 L/min while output at 100 days (n = 2) averaged 4.22 L/min. The electrical current did not change across time during the study, indicating normal operation of the bearings. Pumps consumed less than 10.5 W of power for all support durations. Hemolysis did not occur; the average daily plasma free hemoglobin varied from 2.0 to 8.0 mg/dl. Evaluation of serum biochemical data showed that implantation of the DeBakey VAD in calves with normal hearts did not impair end organ function; BUN, creatinine, and total bilirubin varied minimally within the normal range. The white blood cell count of implanted animals remained within the normal range throughout the study. PMID- 10463512 TI - Symptomatic pharmacotherapy of migraine. AB - This review summarizes data on the effectiveness of various symptomatic migraine pharmacotherapies and makes recommendations for treatment. A wide variety of agents are available for the symptomatic treatment of migraine headache, including over-the-counter analgesics, nonsteroidal anti-inflammatory drugs (NSAIDs), combination products, opiates, ergot alkaloids, corticosteroids, dopamine antagonists, and triptans. In the stepped-care approach, simple analgesics and NSAIDs are the recommended first step for the treatment of mild-to moderate migraine headaches. Patients who do not respond to first-step treatments may be given ergots, combination products, dopamine antagonists, or triptans as the second step. Corticosteroids or opiates may be used as rescue treatment in patients who do not respond to second-step treatment. A stratified approach to care individualizes treatment based on the severity of the headache and other patient-specific factors. In a stratified approach, dihydroergotamine or triptans may be the first-step treatment for patients who present with a history of severe migraines that have responded poorly to previous treatments. Sumatriptan was the first triptan approved for the symptomatic treatment of migraine headache; newer triptans include zolmitriptan, naratriptan, and rizatriptan. Since sumatriptan is rapidly absorbed by the subcutaneous route, its time to onset of effect is shortest. Among triptan drugs that are administered orally, the relative time to onset may be shorter with rizatriptan than sumatriptan. Naratriptan has a longer time to onset but is associated with a lower rate of migraine recurrence than other triptans. graine headache, ergot alkaloids, triptans, PMID- 10463513 TI - Selective cyclooxygenase-2 inhibitors for the treatment of arthritis. AB - The purpose of this paper is to review the rationale for a new class of nonsteroidal anti-inflammatory drugs (NSAIDs) known as selective cyclooxygenase (COX)-2 inhibitors and to present preliminary clinical data on 2 COX-2 inhibitors that are approved for use in the United States. The primary mechanism of NSAIDs in the treatment of inflammation is the inhibition of COX, which exists in 2 forms. COX-I appears to regulate many normal physiologic functions, and COX-2 mediates the inflammatory response. Theoretically, an NSAID that inhibits COX-2 selectively should decrease inflammation but not influence normal physiologic functions and thus should cause fewer gastrointestinal side effects. Preliminary data suggest that celecoxib, a highly selective COX-2 inhibitor, is superior to placebo and similar to traditional NSAIDs in the short-term treatment of pain due to osteoarthritis, although it has been associated with adverse effects such as headache, change in bowel habits, abdominal discomfort, and dizziness. Celecoxib also has been shown to be as effective as traditional NSAIDs in the treatment of rheumatoid arthritis, but it may cause fewer adverse effects, including endoscopically documented ulcers. Celecoxib is metabolized in the liver by the cytochrome P-450 isozyme CYP2C9, and thus serious drug interactions are possible. In the treatment of osteoarthritis, rofecoxib has been shown to be as effective as traditional NSAIDs and may cause fewer endoscopically documented ulcers, but its complete adverse-effect profile is not known. Until the selective COX-2 inhibitors are widely used and more clinical as well as pharmacoeconomic studies are published, the exact role of COX-2 therapy cannot be determined. words: cyclooxygenase, celecoxib, rofecoxib, rheumatoid arthritis, osteoarthritis. PMID- 10463514 TI - Comparison of cefuroxime axetil and amoxicillin/clavulanate in the treatment of acute bacterial sinusitis. AB - This double-masked, multicenter, randomized clinical trial compared the efficacy and tolerability of cefuroxime axetil and amoxicillin/clavulanate in the treatment of acute bacterial maxillary sinusitis. A total of 263 patients with acute bacterial maxillary sinusitis were randomly assigned to receive 10 days of treatment with either cefuroxime axetil 250 mg twice daily (n = 132) or amoxicillin/clavulanate 500/125 mg 3 times daily (n = 131). Patients' responses to treatment were assessed once during treatment (6 to 8 days after the start of treatment), at the end of treatment (1 to 3 days posttreatment), and at follow-up (26 to 30 days after cessation of treatment). Clinical success, defined as cure or improvement, was equivalent in the cefuroxime axetil and amoxicillin/ clavulanate groups at the end-of-treatment and follow-up assessments. Patients in both groups showed improvements in symptoms of acute sinusitis at the during treatment visit. Treatment with amoxicillin/clavulanate was associated with a significantly higher incidence of drug-related adverse events than treatment with cefuroxime axetil (29% vs 17%), primarily reflecting a higher incidence of gastrointestinal adverse events (23% vs 11%), particularly diarrhea. Two patients in the cefuroxime axetil group and 8 patients in the amoxicillin/clavulanate group withdrew from the study due to adverse events (P = 0.06). These results indicate that cefuroxime axetil 250 mg twice daily is as effective as amoxicillin/clavulanate 500 mg 3 times daily in the treatment of acute sinusitis and produces fewer gastrointestinal adverse events. cefuroxime axetil, amoxicillin/clavulanate, acute sinusitis. PMID- 10463515 TI - The cardiac pharmacodynamics of therapeutic doses of sparfloxacin. AB - This double-masked, randomized, placebo-controlled study assessed the cardiac safety of sparfloxacin (as measured by the effect on corrected QT [QTc] interval) at the extremes of the expected therapeutic dosage range. Ninety healthy adult male volunteers with no clinically relevant electrocardiographic (ECG) abnormalities received either placebo or 1 of 3 sparfloxacin regimens consisting of a loading dose on day 1 followed by 3 days of daily dosing at half the loading dose (200/100 mg, 400/200 mg, or 800/400 mg). After each dose, serial blood samples and ECG measurements were obtained to determine the pharmacokinetic and pharmacodynamic variables for sparfloxacin. Increases in the area under the plasma concentration-time curve from time 0 to 24 hours (AUC0-24) for each dosing interval and in the maximum concentration (Cmax) on days 1 and 4 were dose proportional. The steady-state (day-4) values were 6% to 16% lower than the day-1 values. At steady state, the time to C ranged from 2.5 to 3.9 hours across all doses and days studied. The half-life ranged from 18.7 to 20.3 hours. Increases in the placebo-adjusted mean change and mean maximum change in QTc interval were dose related. The placebo-adjusted increases on day 1 were 9, 16, and 28 milliseconds after receipt of the 200/100-mg, 400/200-mg, and 800/400-mg regimens, respectively. The corresponding increases on day 4 were 7, 12, and 26 milliseconds. The placebo-adjusted changes in QTc interval also showed a linear relationship with the AUC0-24 and Cmax of sparfloxacin. In the majority of volunteers (>90%), these increases were within the normal range for the QTc interval (< or = 460 milliseconds). PMID- 10463516 TI - Relation of peripheral neuropathy to HIV treatment in four randomized clinical trials including didanosine. AB - Peripheral neuropathy has been recognized as a dose-limiting adverse effect in Phase I studies of didanosine (ddI) therapy for HIV infection. To study the effect of the currently recommended lower dose of ddI, the databases of 4 randomized, controlled trials were used to assess the frequency of dose-limiting peripheral neuropathy during treatment with ddI 500 or 750 mg/d, compared with zidovudine (ZDV) monotherapy or combination therapy with ddI/ZDV or zalcitabine/ZDV. No between-group differences in risk factors for neuropathy (eg, infectious and metabolic factors, malignancy, concurrent medications) were observed in the individual trials, and the presence of these risk factors appeared to have no increased treatment effect on the occurrence of neuropathy. No significant between-group differences were observed in the individual studies with regard to the incidence or time to onset of peripheral neuropathy. Analysis of the combined results by treatment regimen showed no significant difference in the incidence of neuropathy between recipients of ddI 500 mg/d, ddI 750 mg/d, or ZDV and no significant difference in the cumulative dose received until the onset of neuropathy between the ddI 500- and 750-mg regimens. Entry CD4+ cell counts were significantly predictive of neuropathy, with each 100-cell/microL decrement associated with a 17% increase in risk (P = 0.002); a CD4+ cell count of <50 cells/microL was highly predictive of neuropathy (P = 0.0001). In summary, the risk for peripheral neuropathy was not increased by treatment with ddI versus comparator regimens or by treatment with ddI at the dosages used in studies conducted more recently than the Phase I trials. Peripheral neuropathy seems more likely to be associated with advanced HIV infection and lower CD4+ cell counts (particularly counts <50 cells/microL) than with ddI therapy at the currently recommended dose. PMID- 10463517 TI - Compliance of pediatric patients with treatment involving antibiotic suspensions: a pilot study. AB - In collaboration with 11 German pediatricians in private practice, this pilot study assessed the treatment compliance of 289 pediatric patients (56.1% male: mean age, 53.9+/-35.6 months) who were given antibiotic suspensions (selection and duration determined by the pediatrician) to treat the following bacterial infections: acute otitis media, 34.6%; group A streptococcal tonsillopharyngitis/scarlet fever, 28.7%; lower respiratory tract infection, 18.3%; sinusitis/sinobronchitis, 9.3%; and other infections, 9.0%. The most frequently used antibiotics were amoxicillin (26.3%), erythromycin estolate (19.0%), penicillin V benzathine (14.2%), and cefaclor (13.5%). Compliance was assessed by means of a standardized telephone interview and a urine test that detects antibacterial activity using a Bacillus subtilis spore suspension. Overall compliance (positive urine test result at the end of the planned treatment period) was 79.6% (230 of 289 patients). Compliance was highest with erythromycin estolate (94.5%), followed by penicillin V benzathine (85.4%), cefaclor (76.9%), and amoxicillin (71.1%). Good compliance was also significantly associated with a patient age of > or =3 years and a treatment duration of > or =7 days. Compliance was not significantly influenced by the underlying bacterial infection. In summary, 20.4% of patients were noncompliant when treated with antibiotic suspensions. PMID- 10463518 TI - Pharmacokinetics of sparfloxacin in patients with renal impairment. AB - Sparfloxacin is a fluoroquinolone antimicrobial agent with a broad spectrum of activity and long elimination half-life. Because its single-dose pharmacokinetics are altered by renal impairment, the present study was undertaken to determine the effects of moderate or severe renal insufficiency on the multidose pharmacokinetic characteristics of and tolerance to sparfloxacin. The pharmacokinetic characteristics of sparfloxacin were assessed in 32 subjects (15 men, 17 women) with (1) normal renal function (creatinine clearance [CLcr]> or = 250 mL/min per 1.73 m2) and a mean age of 52.6 years and mean weight of 70.4 kg; (2) moderate renal insufficiency (CLcr 30-49 mL/min per 1.73 m2) and a mean age of 54.4 years and mean weight of 67.8 kg; and (3) severe renal insufficiency (CLcr 10-29 mL/min per 1.73 m2) and a mean age of 50.8 years and mean weight of 73.1 kg. The first 2 groups received a 400-mg loading dose on day 1 followed by 200 mg once daily for 9 days; subjects with severe renal insufficiency received a 400-mg loading dose on day 1 followed by 200 mg every 48 hours on days 3, 5, 7, and 9. The plasma and urinary pharmacokinetics of sparfloxacin and its glucuronide metabolite were determined after the last dose. All subjects were monitored for changes in the corrected QT (QTc) interval and for adverse events. Renal insufficiency altered the steady-state pharmacokinetic variables of sparfloxacin and its glucuronide metabolite, reducing their renal clearances and increasing both maximum plasma concentration and area under the plasma concentration-time curve. Mean steady-state plasma sparfloxacin concentrations in subjects with severe renal insufficiency (48-hour dosing interval) were comparable to those in subjects with normal renal function (24-hour dosing interval). However, mean plasma sparfloxacin concentrations in patients with moderate renal insufficiency were 2 to 3 times greater than the corresponding concentrations in subjects with normal renal function receiving the same dosage regimen. The QTc interval was slightly increased in all groups (the greatest increases were 14, 14, and 6 milliseconds in the groups with normal renal function and moderately and severely impaired renal function, respectively, at 5.5 hours post-dose on day 9 or 10) but similar among subjects with normal renal function or with renal insufficiency. Sparfloxacin was well tolerated. Thus sparfloxacin clearance is reduced and plasma concentrations raised by moderate or severe renal insufficiency. These increases do not appear to augment drug effects on the QTc interval or enhance the risk for adverse events. These results suggest that alternate-day dosing (48-hour dosing interval) following a double loading dose on day 1 should be used in patients with severe renal insufficiency and may be appropriate for patients with moderate renal insufficiency. PMID- 10463519 TI - Intravenous ondansetron for the control of opioid-induced nausea and vomiting. International S3AA3013 Study Group. AB - This randomized, double-masked, placebo-controlled, multicenter trial was conducted in 9 countries to assess the safety and efficacy of 2 doses of intravenous ondansetron (8 and 16 mg) for the control of opioid-induced nausea and vomiting. A total of 2574 nonsurgical patients who presented with pain requiring treatment with an opioid analgesic agent participated in this trial. The most common presenting painful condition was back or neck pain, reported by approximately one third of patients. A total of 520 patients (317 females, 203 males) developed nausea or vomiting after opioid administration and were randomly assigned to receive a single dose of 1 of 3 study treatments: placebo (n = 94), ondansetron 8 mg (n = 215), or ondansetron 16 mg (n = 211). Ondansetron 8 and 16 mg led to complete control of emesis in 134 of 215 patients (62.3%) and 145 of 211 patients (68.7%), respectively. Results with both doses were significantly better than those seen with placebo (43 of 94 patients [45.7%]). Complete control of nausea was achieved in 6.8% of placebo patients, 14.8% of ondansetron 8-mg treated patients, and 19.4% of ondansetron 16-mg treated patients; only ondansetron 16 mg was significantly better than placebo (P = 0.007). Significantly more patients who received ondansetron 8 mg than patients who received placebo were satisfied/very satisfied with their antiemetic treatment, as assessed by 4 patient-satisfaction questions. Significantly more patients who received ondansetron 16 mg compared with placebo were satisfied/very satisfied on 2 of 4 satisfaction questions. In conclusion, based on the observed incidence of opioid-induced nausea and vomiting in this study, it may be more appropriate to treat symptoms on occurrence rather than administering antiemetic agents prophylactically. The results of this study demonstrate that intravenous ondansetron in doses of 8 or 16 mg is an effective antiemetic agent for the control of opioid-induced nausea and vomiting in nonsurgical patients requiring opioid analgesia for pain. PMID- 10463521 TI - Factors contributing to trends in prescription drug expenditures. AB - Between 1970 and 1995, national prescription drug expenditures and Medicaid drug expenditures increased proportionately less than did total health care expenditures and total Medicaid expenditures, respectively, although they increased to a greater extent than did expenditures in other sectors of the economy. General inflation, which cannot be controlled by health care policy, has been the major factor contributing to the growth in national prescription drug expenditures. Other contributors were population growth, increases in per capita prescription use, increases in per-prescription intensity (ie, real drug expenditures), and the fact that prescription drug prices exceeded general inflation. Medicaid drug expenditures have increased mainly because of growth in the number of drug recipients, increases in prescription drug prices, and economy wide inflation. PMID- 10463520 TI - The cost-effectiveness of donepezil therapy in Swedish patients with Alzheimer's disease: a Markov model. AB - This study compared the cost-effectiveness of donepezil, a new cholinesterase inhibitor indicated for the treatment of mild-to-moderate probable Alzheimer's disease (AD), with no treatment. A Markov state transition model was employed to simulate treatment costs based on Swedish epidemiologic data. The Markov states used in the model were defined according to cognitive function, as assessed by the Mini-Mental State Examination. Data on costs and baseline transition probabilities were taken from the Kungsholmen Project, an observational, population-based study of persons aged >75 years in Sweden. Data on the treatment effect were taken from a clinical trial comparing donepezil to placebo over 24 weeks and were applied to the baseline transition probabilities to assess the effect of treating the clinical manifestations of AD in Swedish patients. Also, a within-trial analysis was performed for comparison, using transition probabilities taken from the clinical study. Both models were run for 5 years in half-year cycles, and both demonstrated various degrees of cost savings and improved effectiveness, as measured by increased time in nonsevere disease states. Thus donepezil had superior cost-effectiveness compared with no treatment. PMID- 10463522 TI - Hospital use and costs among patients with nonischemic cardiomyopathy in the first prospective randomized amlodipine survival evaluation study. AB - The incidence of hospitalizations, lengths of stay, and per-diem costs were determined for 421 patients (amlodipine, 209; placebo, 212) with nonischemic cardiomyopathy in the first Prospective Randomized Amlodipine Survival Evaluation (PRAISE) study to assess the impact of amlodipine on hospital use and to compare the costs of hospitalization with the cost of amlodipine treatment. Treatment with amlodipine versus placebo significantly delayed the mean (+/- SD) time to first hospitalization (447 +/- 26 d vs 315 +/- 18 d, respectively; P = 0.0139). Both treatment groups showed a similar number of hospital admissions per patient per year. The overall hospital length of stay was 1.17 days less per year with amlodipine than with placebo, at a cost of $1098 less per person per year although these differences were not statistically significant. Significantly fewer amlodipine patients were admitted for unexplained cardiac arrest (odds ratio, 0.235; P = 0.002) and ventricular arrhythmias (odds ratio, 0.497; P = 0.004). These findings are consistent with clinical reports from PRAISE of prolonged survival and a reduction in sudden cardiac death among patients with severe heart failure due to nonischemic heart disease. This analysis suggests that in patients with nonischemic cardiomyopathy, treatment with amlodipine can delay the time to hospitalization and may reduce the number of hospital admissions related to ventricular arrhythmias. The estimated reduction in hospital costs of $1098 per year would more than offset the amlodipine treatment cost of approximately $700 per year. PMID- 10463523 TI - Market factors and the availability of community pharmacies. AB - The purpose of this study was to examine the relationships between the availability of community pharmacies and 4 types of market factors. A composite data set was created that linked, at the county level, data on: (1) type and number of pharmacies; (2) population characteristics; (3) payer variables; (4) health care system factors; and (5) competitive factors. In this exploratory study, secondary data were used to assess the association between the availability of community pharmacies and the influence of market factors. To assess the market influences on availability of community pharmacies, 2 regressions were performed. In 1 model, the number of community pharmacies per 10,000 population was the dependent variable, whereas the dependent variable in the other regression was the proportion of independently owned community pharmacies. The independent variables in each regression were the market factors- population characteristics, payer variables, health care system factors, and competitive variables. Squared terms were included for 8 of 15 market factors to account for nonlinearities in the relationships. Multiple market factors were correlated with both the number of community pharmacies and the proportion of independently owned pharmacies in an area. Several of the relationships were not linear and changed direction within the range of data. Counties with either a low or a high percentage of elderly people had fewer pharmacies and a lower proportion of independently owned pharmacies compared with counties with a moderate percentage of elderly people. Counties that were scarcely or highly rural had fewer community pharmacies but a higher proportion of independently owned pharmacies than counties that were moderately rural. Areas with a greater percentage of the population earning less than the poverty level had more pharmacies, especially independently owned ones. Fewer community pharmacies were found in areas with higher health maintenance organization penetration rates. The number of hospital admissions was positively associated with the number of pharmacies but negatively associated with the proportion of independently owned pharmacies. The availability of community pharmacies varies across the country. In light of the trend toward fewer independently owned pharmacies, potential problems in accessing pharmacy services could develop in certain areas, including those that are highly rural and those with a high percentage of people earning less than the poverty level. Future research and policy issues are identified. PMID- 10463524 TI - Ten guiding principles for teaching children and adolescents about medicines. US Pharmacopeia. AB - In 1996, an open conference sponsored by the US Pharmacopeia (USP) and attended by more than 100 health care professionals established the need and rationale for teaching children and adolescents about medicines. After the conference, a public, iterative, consensus-development process including participation by 35 health-professional organizations was undertaken. This process resulted in a USP position statement, "Ten Guiding Principles for Teaching Children and Adolescents About Medicines," which supports the right of children and adolescents to receive developmentally appropriate information and direct communications about medicines that are consistent with their health status, capabilities, and culture. The position statement is intended to stimulate activities that will help children become active participants in the process of appropriate use of medicines and prepare them for the day they begin to use medicines independently. PMID- 10463525 TI - Analytical characterisation of cytokines and growth factors. AB - Cytokines (including growth factors) are playing an ever-increasing role in the therapy of human disease. The rapidity by which new cytokines are discovered, cloned and enter the clinics has placed a burden on control authorities and manufacturers to ensure their safety, quality and efficacy. The appropriate characterisation of these proteins plays a vital role in ensuring the development of cytokines as useful therapeutic agents. Highly sophisticated physicochemical techniques exist that can produce specific information about the structure and composition of cytokines including methods such as nuclear magnetic resonance (NMR) and mass spectrometry. However, even a bank of such techniques cannot yet predict the biological activity of cytokines. Biological assays are essential to assess the potency of cytokines and can take several forms, from in vivo assays to in vitro cell-line bioassays and more recently biochemically-based assays. Bioassays require a biological reference standard to define an appropriate unit that can be used to assess the potency of a biological therapeutic. However, bioassay-derived potency is a quality issue and should not have an impact on the perceived efficacy of a biological therapeutic. PMID- 10463526 TI - Cytokine bioassays: an overview. AB - Although eclipsed in recent years by immunoassays and molecular biology techniques, bioassays remain a vital research tool for cytokine biology. Like any type of biological system, cytokine bioassays require meticulous technique to obtain accurate and reproducible results. However, these minor difficulties are more than compensated for by their exclusive detection of biologically active molecules, a feature as yet unmatched by other assay methods. PMID- 10463527 TI - Bioassays for growth factors. AB - Growth factors have a wide variety of actions in living systems, providing a range of potentially quantifiable responses for measurement of their biological activity. The biological activity has to be assessed by bioassay as it cannot be predicted from physicochemical data alone. Bioassay systems range from in vivo responses to changes in receptor binding and in early components of signal transduction pathways. The most commonly used systems are based on the measurement of responses of immortalized cell lines, which although not as functionally relevant as in vivo assays, are easier to use. Most growth factors have multiple actions on multiple targets, and can show differential changes in their different activities, so use of the biological activity measured in one bioassay system to predict biological activity in another system must be rigorously validated. Since the bioassay systems are themselves inherently variable, measurement of the growth factor's activity must be made relative to a common, stable, reference preparation to permit valid inter-assay and inter laboratory comparisons. PMID- 10463528 TI - The immunoassay of cytokines and growth factors in biological fluids. AB - There are a number of problems associated with the development of standards suitable for use in the most commonly used assays to detect cytokines in biological fluids. These problems include: (i) the failure of some MoAbs used in immunoassays to detect all different <> of recombinant or natural material; (ii) the use of many different MoAbs, with different specificities, in different immunoassay kits, and (iii) the detection of non-active cytokines (fragments, inhibitors, receptor antagonists, etc.) in these immunoassays. As a result, it is possible to have biologically active material which is not detected in these immunoassays. Alternatively, biologically inactive material can be detected in these assays and is indistinguishable from biologically active material. In addition, the use of different antibodies with different specificities, affinities and avidities in different kits designed to detect the same biological materials results in markedly different sensitivities and specificities. Many of these same concerns can be raised for the use of bioassays for detection of molecules in biological fluids. The solution will not be simple (if possible at all). In most cases, the immunoassay kits are designed to detect <> material in biological fluids, but are made with MoAbs against recombinant material. Because of the markedly different specificities, affinities, etc. of the MoAbs in these kits, their standardization is possible only with a highly purified preparation of natural material. For the assay of recombinant materials, immunoassays should be specifically designed with the recombinant material in mind (i.e. the MoAbs made specifically against the recombinant material to be detected or shown to bind effectively with the recombinant material). Importantly, it should be made clear to investigators using different immunoassays that: (i) the reporting of biological material detected using immunoassays can only be made in units of weight (i.e. ng/ml); (ii) because of the detection of biologically active and inactive material using immunoassay kits these assays cannot be directly compared to bioassays or their results represented as <>; (iii) because of the difference in specificity and sensitivity of the different reagents used in different immunoassays, the results from different assays cannot be directly compared, and (iv) because of these same considerations, comparison of different > of materials within a single immunoassay is also not possible. The use of specific immunoassays for recombinant material in combination with bioassays and the use of cytokine standards, made from highly purified natural material, would help to standardize the results in this field. PMID- 10463529 TI - Chromatography of recombinant proteins. AB - Variants of intact polypeptides/proteins ranging in mass from 6,500 to 70,000 Da were easily separated using reversed-phaseHPLC (rpHPLC) or affinity chromatography. A variant of rhlGF-I, where the racemization of a serine residue was detected in the intact molecule, was resolved from rhlGF-I within 25 minutes by rpHPLC. Other variants of rhlGF-I separated by this method include methionine sulphoxide at position 59, des Gly1, des Gly1Pro2, Glu for Asp substitution at position 20 and incorrectly folded IGF-I. For rhDNase (approximately 40 kDa), affinity chromatography was able to clearly resolve three different amino acids (Asn, Asp and iso-Asp) at position 74 of the intact glycoprotein. The presence or absence of O-linked sugars on Thr -37 of recombinant human thrombopoietin was rapidly demonstrated by rpHPLC. While the separation of these types of variants is essential, the demonstration of biological activity is critical for designing specifications that allow the administration of these proteins into humans. Once a correlation exists between the variant and its biological activity, control of the manufacturing process can be better achieved with analytical methodology. PMID- 10463530 TI - Spectroscopic characterization of chemokines: relevance for quality control and standardization. AB - Chemokines are mediators of inflammation and trafficking of cells of the immune system including a pivotal role in the recruitment and activation of leukocytes. Due to their involvement in a variety of disease processes, chemokines are potential therapeutic targets. The use of chemokines as pharmaceuticals will require that the folded state and the association properties of the protein are well characterized. In this report, we describe the utility of nuclear magnetic resonance spectroscopy as a tool to study these aspects of chemokine structural properties. PMID- 10463531 TI - Bioassays for the characterisation and control of therapeutic cytokines; determination of potency. AB - The primary value of bioassays is that they alone directly assess the biological activity of bioactive substances and products like cytokines. Appropriately designed bioassays reflect the fundamental aspects of the biological activity of a cytokine molecule, including ligand-receptor binding, signal transduction processes (often poorly understood) and the final observed biological effects. Biological assays therefore complement physicochemical and biochemical procedures which normally only assess precise molecular structural features of cytokines. Bioassays provide valuable information concerning the potency of cytokine products. This is essential for evaluating batch-to-batch consistency, appropriate formulations and stability. Bioassay data are crucial at all stages in the development of cytokine products, from early research to final quality control of finished product. However, the type and design of bioassays may differ according to the information required and its intended use. The assays may or may not directly relate to the clinical use of the product. Bioassays can be difficult to perform and time consuming, although this often reflects bad assay choice and/or design. Correct analysis of the assay results is essential if valid data are to be obtained. Standardisation, using correctly calibrated primary and secondary standards, is essential. In vivo bioassays are normally more unreliable than in vitro procedures, but in some cases in vitro systems are either not available or do not address important biological characteristics of a product. Bioassays must be validated for their intended purpose and for the types of samples to be measured. Appropriate statistical analysis should be used to derive the significance and specifications of results. This needs to address both variability in samples and assay performance. Specifications (limits) for product acceptability need to be derived from real data using several batches of cytokine product. PMID- 10463532 TI - Bioassays and biological stability. AB - The development of the concept of <>, made possible through the power and detail afforded by modern biochemical and biophysical techniques, has resulted in questions being raised about the value of the more variable bioassay. However bioassays, particularly in a routine stability monitoring situation, continue to be useful and often provide unique information not obtained by other techniques. They have advantages in that they are relatively easy to perform and generally have better sensitivity than most structural techniques. They can be used to monitor for previously undetected changes - particularly those associated with conformational alterations, and can be used to monitor the effective combination of all individual changes. PMID- 10463533 TI - A comparison of physico-chemical and biological assays in the batch release of therapeutic cytokines. PMID- 10463534 TI - The role of the bioassay in the evaluation of cytokines and growth factors for regulatory decision making. PMID- 10463535 TI - Novel pathways for negative regulation of inflammatory cytokines centered on receptor expression. PMID- 10463536 TI - Blocking HIV co-receptors by chemokines. AB - Specific chemokines can block HIV entry and replication because they antagonize the common strategy of lentiviruses to use chemokine receptors for infecting CD4+ cells of the body, especially lymphocytes and cells of the monocytic lineage. This raised intense academical and therapeutical interest. The antiviral potency of these chemokines is indeed remarkable, but depends on the chemokine and the HIV isolate used. This is because HIV appears to use many co-receptors, alternatively or in addition to the CCR5 co-receptor. These include CCR3, CXCR4, STRL33/Bonzo/TYMSTR, and BOB. The CC chemokines RANTES, MIP-1alpha, MIP-1beta, and Eotaxin can suppress the replication of CCR5- and CCR3-dependent viruses, while SDF-1 alpha/beta suppresses that of CXCR4-dependent strains. Although no general rule can be drawn at present, it appears that chronic HIV infection may give rise to viruses which, instead of using preferentially or exclusively CCR5, are capable of using more than one co-receptor. This underlines the need for assaying the tropism of primary isolates, using both fusion assays and protection of activated lymphocyte cultures by one or more antiviral chemokines or chemokine antagonists. PMID- 10463537 TI - How much insulin-like growth factor-I (IGF-I) circulates?: impact of standardization on IGF-I assay accuracy. AB - Insulin-like growth factor-I (IGF-I, somatomedin-C) is a peptide hormone which plays an important role in growth regulation. Accurate measurements of circulating IGF-I levels in plasma samples are an important part of many studies on growth and development, and therefore many assays have been developed for this purpose. We have found that assay standardization has a major impact on IGF-I quantification. Most IGF-I assays are calibrated against the World Health Organization (WHO) International Reference Reagent (IRR) for IGF-I Immunoassays (87/518). The protein content assigned to WHO IRR 87/518 was a consensus value from a multicentre WHO collaborative study. Here we present physico-chemical data showing that WHO IRR 87/518 is Met(-1) IGF-I of low purity (44%), and that the assigned protein content is somewhat higher than the <> value determined by quantitative amino acid analysis. We show that assays calibrated against WHO IRR 87/518 report total IGF-I concentrations that are in excess of actual values by approximately two-fold. For this reason much of the IGF-I concentration data in the literature, which are based on assays calibrated against WHO IRR 87/518, are of questionable accuracy. PMID- 10463538 TI - Kinase Receptor Activation (KIRA): a rapid and accurate alternative to endpoint bioassays. AB - We have developed a novel strategy for a rapid bioassay that is accurate, precise, sensitive, and high capacity. It is capable of quantifying ligand bioactivity by measuring ligand-induced receptor tyrosine kinase activation in terms of receptor-phosphorylation. The assay, termed << Kinase Receptor Activation>> or KIRA, uses two separate microtiter plates, one for ligand stimulation of intact cells, and the other for receptor capture and phosphotyrosine ELISA. The assay makes use of either endogenously expressed receptors or stably transfected receptors with a polypeptide flag. KIRA assays for the ligands IGF-I and NGF were compared to their corresponding endpoint bioassays (3T3 cell proliferation for IGF-I and PC12 cell survival for NGF). The KIRA assays showed excellent correlation with the more classical endpoint bioassays. Further, they were highly reproducible, minimizing the requirement for repeat assays. The KIRA assay format has great potential as a rapid, accurate and precise bioassay, both for potency determination as well as stability-indicating analyses. PMID- 10463540 TI - Statistical validity of bioassays. PMID- 10463539 TI - Applications for the new electrochemiluminescent (ECL) and biosensor technologies. AB - Biosensor and electrochemiluminescent (ECL) assays are replacing enzyme-linked immunosorbent assays (ELISAs) at Schering-Plough as immunoassays of choice to monitor cytokine levels and detect anti-cytokine antibody responses during cytokine therapy. These new assays provide increased sensitivity and a better correlation with biological assays. Biosensor assays using the BIACORE 2000 (BIACORE, Uppsala, Sweden) are being adopted to support preclinical and clinical trials for the detection of antibodies capable of binding to IL-10 and IL-4. Significant advantages when using a biosensor assay are that real-time and label free detection permit increased throughput and direct detection of binding interactions which enables detection of low affinity antibodies that are not detected by ELISA. The ECL assays using the ORIGEN Analyser (IGEN, Gaithersburg, MD) that we have implemented to replace existing ELISAs for quantification of serum IL-10 and serum interferon alfa levels are more sensitive and less subject to matrix effects. Data obtained during the validation of these assays are described. PMID- 10463541 TI - Validation of bioassays for quality control. AB - For a biological assay to be useful for quality control it should fail bad lots, pass good lots, and estimate relative potency with high accuracy and precision. To fail a lot we rely most heavily on the test for parallelism. For the parallelism test and other preliminary tests as well as for inference, appropriate estimates of assay variation are crucial. Location effects on 96 well plates and serial dilution of samples using multichannel pipettes make it difficult to obtain good estimates of assay variation. This paper develops the use of a split-block design and analysis where blocks are reasonably consistent regions of a plate; this approach removes some location effects, allows other location effects to be treated as assay variation and provides appropriate measures of assay variation. Randomization, even within the split-block design, is difficult without robots to reduce the likelihood of procedural errors. There are hardware, software, and validation obstacles to implementation of robots in the bioassay laboratory. More generally, validation of a bioassay should be reported on log relative potency and must address between- and within-assay variation. When between assay variation is not small, the usual weighted approach to combining relative potency estimates (which ignores between-assay variation) is inappropriate; a simple sampling average and standard deviation is a better solution. PMID- 10463542 TI - Biological standardization of cytokines and growth factors. AB - To quantify the biological activity of different cytokine preparations with different specific activities by bioassay, mass units cannot be used and biological activity has to be expressed as << biological potency units >>. The biological unit requires definition by a standard that is assay-independent (especially when measuring a particular type of biological activity). Once the unit is defined, it can be used in any laboratory, thus providing a means of ensuring uniformity throughout the world in the designation of potency of different biological preparations. The World Health Organisation (WHO) standardization programme involves the production of biologically stable, well characterised potency and immunoassay standards that are available world-wide using a single international unitage. These international standards have been used to reduce dramatically the variation in estimates of cytokine preparations within and between laboratories for immunoassays and bioassays. PMID- 10463543 TI - The role of Pharmacopoeia standards and monographs. PMID- 10463544 TI - Emergency medicine's "illusion of efficacy" and the public perception of resuscitation research. PMID- 10463545 TI - Can acutely ill patients consent to research? Resolving an ethical dilemma with facts. PMID- 10463546 TI - Informed consent: the most important protector. PMID- 10463547 TI - Do patients with acute medical conditions have the capacity to give informed consent for emergency medicine research? AB - OBJECTIVE: Because of stress and illness, conscious emergency medicine (EM) patients may be temporarily cognitively impaired and thus incapable of participating in the informed consent process for acute care research. This pilot study sought to assess the mental capacity of ED patients during their evaluation and treatment for acute myocardial infarction (AMI). METHODS: A prospective observational design at a university tertiary referral center. EM patients with AMI from November 1996 to February 1997 were enrolled. While usual care was delivered, patients were administered three subtests of the Wechsler Adult Intelligence Scale-Revised (WAIS-R) test. Subtest scaled scores range from 1 to 19, with abnormal being less than 5. Demographic, historical, and environmental parameters were recorded. Patients assessed how serious they perceived their conditions; and rated their degrees of pain, nausea, breathlessness, and anxiety on 10-cm visual analog scales (VASs). Testing was repeated prior to hospital discharge. RESULTS: Twenty-five patients were enrolled. Of these, two (8%) were suspected by their emergency physicians to have insufficient capacity to give informed consent. However, five (20%) scored less than 5 on all of the WAIS-R subtests (kappa = 0.5) and eight (32%) scored less than 5 on at least one of the subtests (kappa = 0.3). The initial median Digit Span, Comprehension, and Similarities subtest scores were 7, 5, and 6. By discharge, these improved to 8, 7, and 8, respectively. CONCLUSIONS: This pilot study suggests that some patients with AMI may have difficulty processing information necessary to give informed consent for acute care research. Routine clinical evaluation may not detect this cognitive defect. PMID- 10463548 TI - Impact of patient acuity on preference for information and autonomy in decision making. AB - OBJECTIVES: To test the hypotheses that ED patients' desires for medical information and for autonomy in decision making are inversely related to increasing acuity of illness, increasing age, and lower level of formal education. METHODS: The authors conducted a prospective study of ED patients who presented for care during seven nonconsecutive 24-hour periods. Of 804 patients approached, 665 completed a questionnaire that was administered by a trained research assistant. Patients rated their desire for medical information, and for participation in medical decision making, on two 10-cm visual analog scales. Patient acuity level was determined in routine fashion by trained triage nurses, who were unaware of the nature of this study. RESULTS: Desire for information was uniformly high, and did not vary statistically between triage groups (p = 0.41). The most acutely ill patients (level I) were more likely to be excluded by the research interviewer (p < 0.001). Of included level I patients, desire to participate was not decreased (p < 0.01). Higher level of formal education (p = 0.036) and younger age (p < 0.001) were associated with greater desire for autonomy in decision making. CONCLUSION: Among ED patients able to participate, higher acuity of illness was not associated with a decreased desire for medical information. Many very acutely ill patients preferred autonomy in medical decision making. Older patients and those with less formal education expressed a lesser desire for decision-making autonomy. PMID- 10463549 TI - History of domestic violence among male patients presenting to an urban emergency department. AB - OBJECTIVE: To establish the prevalence of domestic violence committed by women against male patients presenting to an urban ED for any reason. METHODS: This was a prospective survey in which male patients of legal age presenting to the ED over a 13-week period were interviewed. Patients answered a series of six questions adapted from the George Washington University Universal Violence Prevention Screening Protocol. Patients who could not speak English, those refusing to participate, those unable to give informed consent, and those meeting regional criteria for major trauma were excluded. RESULTS: Of 866 male patients interviewed, 109 (12.6%) had been the victims of domestic violence committed by a female intimate partner within the preceding year. Victims were more likely to be younger, single, African American, and uninsured. The most common forms of assault were slapping, grabbing, and shoving (60.6% of victims). These were followed by choking, kicking, biting, and punching (48.6%), or throwing an object at the victim (46.8%). Thirty-seven percent of cases involved a weapon. Seven percent of victims described being forced to have sex. Nineteen percent of victims contacted the police; 14% required medical attention; 11% pressed charges or sought a restraining order; and 6% pursued follow-up counseling. CONCLUSIONS: Almost 13% of men in this sample population had been victims of domestic violence committed by a female intimate partner within the previous year. Further attention to the recognition and management of domestic violence committed by women against men may be warranted. PMID- 10463550 TI - Emergency department characteristics of male sexual assault. AB - OBJECTIVE: To characterize the nature, frequency, and treatment of male sexual assault encountered by physicians in an ED. A minor objective compared the lengths of time the victim knew the assailant between males and females to determine whether there were differences between male and female victims. METHODS: Retrospective review over four years of all male patients with a diagnosis of sexual assault presenting to an urban academic ED in New York City. Demographics, types of injury, assailant/victim information, and treatment of the assault were obtained. RESULTS: Twenty-seven male sexual assaults (approximately 12% of all sexual assaults) were documented during this time period. Forcible rectal, oral, or both rectal and oral intercourse occurred 14, 4, and 9 times, respectively. Documented physical trauma occurred in nine of 27 visits, with two admissions for head trauma. Five additional patients complained of rectal pain with no independent objective evidence of trauma. Prophylactic treatment with antibiotics for the prevention of Neisseria gonorrhoeae and Chlamydia trachomatis was offered in 26 of 27 episodes and prophylactic HIV protection was given to only two of 21 eligible patients. Documentation of HIV status and HIV counseling occurred only in ten and five visits, respectively. The male-to-female odds ratios for whether the assailant was unknown, known less than 24 hours, or known more than 24 hours were 0.43 (95% CI = 0.15 to 1.26), 2.48 (95% CI = 0.94 to 6.53), and 0.92 (95% CI = 0.31 to 2.71), respectively. CONCLUSIONS: The percentage of total sexual assaults occurring in males who present to an ED is now more than 10% of all sexual assaults seen in this urban ED. These assaults are associated with a high proportion of patients with documented trauma. Although the treatment of traditional sexually transmitted diseases appears to be covered well, the need for HIV documentation, counseling, and possible HIV prophylaxis should be addressed more aggressively. In comparing the lengths of the relationships between the victim and the assailant, it appears twice as likely that the female victim will not know her assailant compared with the male, and twice as likely that the male victim will know his assailant less than 24 hours compared with the female. PMID- 10463551 TI - Identification of intra-abdominal injuries in children hospitalized following blunt torso trauma. AB - OBJECTIVE: To determine the utility of the ED physical examination and laboratory analysis in screening hospitalized pediatric blunt trauma patients for intra abdominal injuries (IAIs). METHODS: The authors reviewed the records of all patients aged <15 years who sustained blunt traumatic injury and were admitted to a Level 1 trauma center over a four-year period. Patients were considered high risk for IAI if they had any of the following at ED presentation: decreased level of consciousness (GCS < 15), abdominal pain, tenderness on abdominal examination, or gross hematuria. Patients without any of these findings were considered moderate risk for LAI. The authors compared moderate-risk patients with and without IAIs with regard to physical examination and laboratory findings obtained in the ED. RESULTS: Of 1,040 children with blunt trauma, 559 (54%) were high-risk and 481 (46%) were moderate-risk for IAI. 126 (23%) of the high-risk and 22 (4.6%) of the moderate-risk patients had IAIs. Among moderate-risk patients with and without IAIs, those with IAIs were more likely to have abdominal abrasions (5/22 vs 34/459, p = 0.008), an abnormal chest examination (11/22 vs 86/457, p = 0.01), higher mean serum concentrations of aspartate aminotransferase (AST) (604 U/L vs 77 U/L, p < 0.001) and alanine aminotransferase (ALT) (276 U/L vs 39 U/L, p = 0.002), higher mean white blood cell (WBC) counts (16.3 K/mm3 vs 12.8 K/mm3, p < 0.001), and a higher prevalence of >5 RBCs/hpf on urinalysis (7/22 vs 54/427, p = 0.02). There was no significant difference (p > 0.05) between moderate-risk patients with and without IAIs in initial serum concentrations of amylase, initial hematocrit, drop in hematocrit >5 percentage points in the ED, or initial serum bicarbonate concentrations. CONCLUSION: In children hospitalized for blunt torso trauma who are at moderate risk for IAI, ED findings of abdominal abrasions, an abnormal chest examination, and microscopic hematuria as well as elevated levels of AST and ALT, and elevated WBC count are associated with IAI. PMID- 10463552 TI - Prevalence of smokers and nicotine-addicted patients in a suburban emergency department. AB - OBJECTIVE: To define the prevalence of smokers and nicotine-addicted patients in a suburban, community ED. METHODS: This was a prospective survey of consecutive ED patients seen in a suburban ED with an annual patient census of 48,000. Medically stable patients aged 18 years or older were eligible for inclusion. Patients were excluded if they had predominantly psychiatric complaints or were critically ill. Patients were queried about their smoking habits by a closed question survey, which included the previously validated Fagerstrom Test for Nicotine Dependence. The study was conducted during a six-week period, only at times when there were dedicated research associates available to ensure consecutive patient entry. Continuous variables were analyzed by Student's t tests. Clinical variables were analyzed by chi-square tests. All tests were two tailed with alpha at 0.05. RESULTS: 1,515 patients comprised the study group. The mean age (+/-SD) was 45.6 (+/-18.9) years; 52% were female, 25% were nonwhite, and 47% were college graduates. There were 317 (21%) smokers. Patients having private physicians were less likely to smoke (18% vs 29%, p = 0.001). Of all smokers, 46% were moderately to severely nicotine-dependent, 69% wanted to quit, and 30% expressed an interest in joining a smoking cessation program. CONCLUSION: A substantial percentage of ED patients smoke, many of them are nicotine addicted, and the majority would like to quit. Randomized, controlled trials are needed to determine whether interventions by physicians in the ED can have an impact on the smoking habits of these patients. PMID- 10463553 TI - The perceived effectiveness of total quality management as a tool for quality improvement in emergency medicine. AB - OBJECTIVES: To describe the perceived effectiveness of using the Total Quality Management (TQM) approach to quality improvement in both academic and nonacademic EDs, and to discuss some important barriers to effectiveness of TQM programs. METHOD: A mail survey of 100 EDs was conducted with telephone follow-up. Hospitals were randomly selected from three subgroups: university teaching hospitals, nonuniversity teaching hospitals, and private nonteaching hospitals. ED physician directors or nonphysician administrators with knowledge of departmental quality improvement initiatives were surveyed. RESULTS: The overall response rate was 60%. Of the respondents, 54 (90.0%) used TQM techniques as part of their quality improvement initiatives. TQM techniques were used more frequently and for a longer duration in academic programs. ED staff participation in TQM projects was relatively low; less than 25% in the majority (79.6%) of all EDs. TQM initiatives were ranked least effective in university settings, of which 11 of 13 (84.6%) rated their TQM programs as ineffective or having no effect. More mature programs (>5 years old) had a significantly higher ranking for effectiveness than those programs less than 2 years old. CONCLUSIONS: Total Quality Management is being utilized in a large number of EDs. TQM initiative is perceived as having little or no positive effect. This is particularly the case in academic EDs. PMID- 10463554 TI - Linking emergency department patients to primary care: phone vs face-to-face interaction. AB - OBJECTIVE: To compare the patient compliances and time analyses of two methods of assigning primary care providers (PCPs) to ED patients who are without a PCP: phone interview vs face-to-face interaction. METHODS: Prospective observational cohort study following an intervention, performed in a pediatric ED, serving a population of 1.7 million, with a census of 80,000 pediatric ED visits per year. Over one year, a consecutive sampling of 1,062 patients evaluated in the ED and without a PCP were approached to participate in our study (536 enrolled, 526 declined). Patients enrolled were addressed by a nurse practitioner/social worker (NP/SW) who arranged an appointment with a PCP, either in person (in ED) or by phone after discharge. The primary outcome measure was compliance with the arranged appointment. Secondarily, the authors analyzed the time necessary for each approach in person-hours. RESULTS: Of the 536 enrolled, 81 were excluded because data collection was incomplete at the time of the study's completion, leaving 455 study patients. Seventy-six percent of the study patients were between the ages of 1 month and 12 years. Contact was made by phone for 151 (33%) patients and face-to-face for 304 (67%). Sixty-two percent of the phone patients kept their appointments, compared with 52% of face-to-face patients (p = 0.048, RR = 1.20, 1.02 < RR < 1.41). Phone interaction was also more time effective. CONCLUSION: Linking ED patients without a medical provider to PCPs via phone is as effective as a face-to-face interaction. PMID- 10463555 TI - A health promotion intervention for families in a Medicaid managed care plan. AB - OBJECTIVE: To determine whether implementation of an intervention based on a model of health promotion will encourage patients to seek care from their primary care provider (PCP) and reduce visits to the pediatric ED (PED) for minor illness. METHODS: Prospective, randomized, controlled study in the PED of an urban children's hospital (CH). Children <13 months old, enrolled in a Medicaid managed care plan, who identified the CH as their site for primary care and presented to the PED for evaluation of minor illness were enrolled after being seen by the triage nurse, before being seen by a physician. Subjects were randomly assigned to the intervention (I) group or control (C) group. Parents of all enrollees completed a survey about health care utilization habits. Each family in the I group received health promotion teaching from a single investigator. The intervention consisted of a review of the child's medical record with the parents, an explanation of what to expect at future well-child visits, and a discussion of the role of the PCP. A follow-up appointment was also provided prior to discharge from the PED. The C group received usual care. Use of health care by all subjects was tracked for one year by medical record review and phone interviews at six and 12 months. RESULTS: 102 subjects in the I group and 93 in the C group (mean +/- SD ages 6.4 months +/- 3.8 and 7.2 months +/- 3.9, respectively, p = 0.15) were enrolled from March 1996 to November 1996. The two groups were similar with respect to demographics and overall health status at enrollment. At study entry: 94 of 102 (92%) subjects in I and 87 of 93 (94%) in C had made at least one visit to the PED in the previous 12 months (p = 0.11); 95 of 102 (93%) in I and 75 of 93 (81%) in C had seen their PCP at least once for well-child care (p = 0.24). Twelve-month follow-up by medical record review was completed for all subjects; phone interviews were completed in 90 of 102 (88%) in I and 80 of 93 (86%) in C. At 12-month follow-up: 84 of 102 (82%) in I and 73 of 93 (78%) in C had made at least one visit to the PED (p = 0.59); 81 of 102 (79%) in I and 77 of 93 (83%) in C had made at least one visit to their CH PCP (p = 0.54). CONCLUSIONS: There was no difference in health care utilization between the intervention and control groups at 12-month follow-up. The health promotion intervention did not alter utilization habits. PMID- 10463556 TI - The integration of primary care sports medicine into an academic emergency medicine practice: academic and revenue enhancement. AB - OBJECTIVES: To determine whether integrating primary care sports medicine into academic emergency medicine (EM) can enhance both revenue and the academic program. METHODS: A retrospective descriptive review of all patients seen in a primary care sports medicine practice at a university hospital sports medicine clinic was done over a 24-month period. All patients seen initially in the ED for a sports injury either by the author or by another EM faculty member with follow up by the author in the sports medicine clinic were included in the study group. The study group was analyzed for diagnoses, payor mix, and revenue generated by the ED follow-up sports medicine clinic visits. RESULTS: There were 199 patients who met the inclusion criteria. This resulted in 483 ED follow-up sports medicine clinic visits. The author practiced 13 hours/week in the ED and 16 hours/week in the primary care sports medicine practice, which resulted in 1,536 sports medicine clinic hours. The study group accounted for 20% of the total patient volume in the author's primary care sports medicine practice. There were 111 lower-extremity injuries (knee 52%, foot/ankle 40%, hip/pelvis 8%), 81 upper extremity injuries (hand/wrist 48%, shoulder 43%, elbow 9%), and seven spine injuries. Payor mix was 47% traditional indemnity, 45% HMO, 4% self-pay, and 4% Medicare/Medicaid. Total charges for the ED follow-up sports medicine clinic visits were $44,767 ($92.68/visit) and net receipts were $30,276 ($62.68/visit). This represented 20% of the total charges and 16% of the net receipts in the author's sports medicine practice during this period. Revenue generated by the ED follow-up sports medicine clinic visits could have supported 12% of the equivalent cost of the base pay for a full-time EM faculty position. CONCLUSION: The integration of primary care sports medicine into an academic EM faculty practice can enhance revenue through the establishment of an ED follow-up sports medicine clinic while also providing an opportunity to expand resident learning experiences. PMID- 10463557 TI - Recent trends and controversies in industry-sponsored clinical trials. AB - In recent years, industry has become an important source of funding for biomedical research. Industry-sponsored clinical trials are a particular source of controversy. In light of recent developments, the authors reevaluate the 1995 SAEM guidelines for investigator involvement in industry-sponsored clinical trials. The authors divide industry-funded clinical trials into two categories: investigator-initiated and industry-initiated, and discuss the differences between them. They examine several areas of ethical debate, including exclusivity contracts between a principal investigator and a corporate sponsor, the size of per-patient reimbursements for recruiting patients into clinical trials, and authorship criteria. Finally, the authors oppose the assumption that industry sponsored research is automatically biased, and suggest that multiple levels of review will help to uncover bias, whatever the source. Once mutual respect for ethical guidelines and practices are established, collaboration between emergency medicine researchers and industry should be encouraged. PMID- 10463558 TI - Research fundamentals VI: misconduct in biomedical research. AB - The goal of medical research is the acquisition and application of new knowledge for the benefit of individual patients and society as a whole. Achieving this goal requires excellence in scientific methodology, honesty in data collection and interpretation, and realistic assessment of the implications of the findings. Underlying all steps in the acquisition of new knowledge is the absolute need for application of the highest ethical standards for research. Occasionally, ethical principles of research are breached because of lack of understanding or the carelessness of researchers. However, researchers have the obligation to know and apply basic principles of research ethics in order to avoid, prevent, or recognize deviations from ethical scientific behavior. When intentional violations of the principles of ethical research occur, the impact to the scientific and lay community can be profound. Misconduct can be prevented if the ethical principles of research are understood and consistently applied. This paper describes the sources and detection of misconduct in the production of science in order to provide emergency researchers with the knowledge needed to prevent misconduct from occurring at all. PMID- 10463559 TI - Evaluation of two strategies for complying with state-mandated lead screening in the emergency department. Naussau-Suffolk Lead Committee, Naussau-Suffolk Lead Center. PMID- 10463560 TI - Poison center data accuracy: a comparison of rural hospital chart data with the TESS database. Toxic Exposure Surveillance System. PMID- 10463561 TI - Domestic violence education in medical school: does it make a difference? PMID- 10463562 TI - Domestic violence among emergency medical technicians. PMID- 10463563 TI - Quadriceps compartment syndrome from minor trauma. PMID- 10463564 TI - Emergency medicine faculty as poison center directors. PMID- 10463565 TI - Reinterpreting physical violence: outcome of intense stimulation-seeking behavior. PMID- 10463566 TI - Caring for the Orthodox Jewish patient on the Sabbath. PMID- 10463567 TI - Airway research. PMID- 10463569 TI - Abnormal regulation of DNA methyltransferase expression during colorectal carcinogenesis. AB - Somatic changes in CpG dinucleotide methylation occur quite commonly in human cancer cell DNA. Relative to DNA from normal human colonic cells, DNA from human colorectal cancer cells typically displays regional CpG dinucleotide hypermethylation amid global CpG dinucleotide hypomethylation. The role of the maintenance DNA methyltransferase (DNMT1) in the acquisition of such abnormal CpG dinucleotide methylation changes in colorectal cancer cells remains controversial; in one study, 60-200-fold increases in DNMT1 mRNA expression were detected in colorectal polyps and cancers relative to normal colonic tissue [W. S. El-Deiry et al., Proc. Natl. Acad. Sci. USA, 88: 3470-3474, 1991], whereas in another study, only small increases in DNMT1 mRNA expression, commensurate with differences in cell proliferation accompanying colonic tumorigenesis, were observed [P. J. Lee et al., Proc. Natl. Acad. Sci. USA, 93: 10366-10370, 1996]. To definitively ascertain whether abnormal DNMT1 expression might accompany human colorectal carcinogenesis, we subjected a series of normal and neoplastic colonic tissues to immunohistochemical staining using a polyclonal antiserum raised against a DNMT1 polypeptide. A concordance of DNMT1 expression with the expression of PCNA and other cell proliferation markers, such as Ki-67 and DNA topoisomerase IIalpha, was observed in normal colonic epithelial cells and in cells comprising other normal epithelia and lymphoid tissues. The polypeptide p21, which has been reported to undermine DNMT1 binding to proliferating cell nuclear antigen at DNA replication sites, was not expressed by normal colonic cells containing DNMT1 and other cell proliferation markers. In adenomatous polyps, although DNMT1 expression coincided with the expression of other cell proliferation markers, many DNMT1-expressing cells also expressed p21. The fidelity of DNMT1 expression was further undermined in colorectal carcinomas, in which a striking heterogeneity in DNMT1 expression, with some carcinoma cells containing very high DNMT1 levels and others containing very low DNMT1 levels, was observed. These results indicate that human colorectal carcinogenesis is accompanied by a progressive dysregulation of DNMT1 expression and suggest that abnormalities in DNMT1 expression may contribute to the abnormal CpG dinucleotide methylation changes characteristic of human colorectal carcinoma cell DNA. PMID- 10463570 TI - Multimodal cancer treatment mediated by a replicating oncolytic virus that delivers the oxazaphosphorine/rat cytochrome P450 2B1 and ganciclovir/herpes simplex virus thymidine kinase gene therapies. AB - Multimodal therapy is generally more effective than single-agent treatment for cancer. rRp450 is an engineered herpes simplex viral mutant that replicates in and kills tumor cells in a relatively selective fashion. It also expresses, in infected cells, the cyclophosphamide (CPA)-sensitive rat cytochrome P450 2B1 (CYP2B1) and the ganciclovir (GCV)-sensitive herpes simplex virus thymidine kinase (HSV-TK) transgenes. We show that cultured rat 9L and human U87deltaEGFR glioma cells, infected and lysed by rRp450, also exhibit supra-additive sensitivity to both CPA and GCV, as determined by Chou-Talalay synergy analysis. DNA cross-linking, assayed by ethidium bromide fluorescence, was significantly inhibited in the presence of GCV, suggesting that interactions between the CPA/CYP2B1 and GCV/HSV-TK gene therapies occurred at the level of DNA repair. In vivo, regression of 9L s.c. tumor volumes in athymic mice was achieved only by the multimodal treatment allowed by rRp450 viral oncolysis combined with CPA/CYP2B1 and GCV/HSV-TK gene therapies, whereas all other treatment combinations produced only tumor growth retardation. PMID- 10463571 TI - FHIT loss of function in human primary breast cancer correlates with advanced stage of the disease. AB - The FHIT gene, encompassing the FRA3B fragile site at chromosome 3p14.2, is a tumor suppressor gene involved in different tumor types. We have assessed 29 human primary breast carcinomas for both the presence of abnormal FHIT transcripts and the Fhit protein levels as compared with the normal breast epithelium of the same patients. In addition, we have also examined a second retrospective series of 156 consecutive breast carcinomas for the expression of the Fhit protein. In nine (31%) cases of the first series, FHIT transcripts were either aberrant or absent as determined by reverse transcription-PCR, and Fhit protein levels in tumors were low or absent as determined immunohistochemically. In 11 other cases (38%), only normal FHIT transcripts were detected by PCR, paralleled by the reduction or absence of Fhit protein. In the remaining nine cases (31%), the presence of the normal FHIT transcript corresponded to protein levels that were similar in tumor and normal breast epithelia. Thus, alterations in FHIT transcripts were detected in 31% of the patients, but reduction or absence of Fhit protein occurred in 69% of the breast carcinoma samples examined. These data suggest that alteration in Fhit expression in breast carcinomas is a frequent event. Analysis of correlation between Fhit expression and pathological, clinical, and biological parameters in these 29 tumors and in a second retrospective series of 156 consecutive primary breast carcinomas indicated that a decrease or an absence of Fhit protein expression is associated with high proliferation and large tumor size. PMID- 10463572 TI - Frequent deletion of hSNF5/INI1, a component of the SWI/SNF complex, in chronic myeloid leukemia. AB - During routine two-fusion fluorescence in situ hybridization analysis of patients with blast crisis of chronic myeloid leukemia (CML), we observed that yeast artificial chromosome 29GD7, which is distal to BCR at 22q11, failed to hybridize to the 9q+ derivative chromosome in 3 of 11 (27%) cases. This deleted region is close to hSNF5/INI1 (SMARCB1), a gene that encodes a widely expressed component of the SWI/SNF chromatin remodeling complex and that suffers biallelic mutations in malignant rhabdoid tumors. To determine whether hSNF5/INI1 was also deleted in patients with CML, we performed fluorescence in situ hybridization analysis with a specific cosmid probe. Deletion of hSNF5/INI1 on the 9q+ chromosome was found in 9 of 25 (36%) cases in blast crisis (lymphoid, n = 3; myeloid, n = 6). For the three of these nine patients for whom material was available prior to transformation, deletions were also seen in chronic phase, indicating that they are early events. Analysis of an additional 21 patients in chronic phase revealed heterozygous loss of hSNF5/INI1 in 5 (24%) cases. Of the 14 patients who had hSNF5/INI1 deletions, 7 showed a mosaic pattern of hybridization in which only a proportion of CML cells that harbored both the t(9;22) derivative chromosomes had a deletion, indicating that loss of hSNF5/INI1 was acquired during the course of the disease. Single-strand conformation polymorphism analysis of all nine hSNF5/INI1 exons and splice junctions failed to reveal any mutations for 31 patients in transformation, including 8 who had deletions, although two polymorphisms were identified. We conclude that deletions of hSNF5/INI1 are frequent in patients with CML. Such deletions may be associated with reduced levels of hSNF5/INI1 expression, which could contribute to leukemogenesis by altering chromatin-mediated transcriptional control. Alternatively, the deletions could target another unidentified gene at 22q11 that plays a role in the pathogenesis of CML. PMID- 10463573 TI - Intestinal dysplasia and adenoma in transgenic mice after overexpression of an activated beta-catenin. AB - Mutations in the adenomatous polyposis coli gene or activating mutations in the beta-catenin gene itself are thought to be responsible for the excessive beta catenin signaling involved in intestinal carcinogenesis. We generated transgenic mice that expressed large amounts of a NH2-terminally truncated mutant beta catenin (deltaN131beta-catenin) in the intestine. These mice had multifocal dysplastic lesions in the small intestine, reminiscent of the early lesions observed in the mouse models of familial adenomatous polyposis. The number of apoptotic cells in the villi of these transgenic mice was 3-4-fold higher than in nontransgenic mice. Expression of the truncated beta-catenin mutant in the kidney led to the development of severe polycystic kidney disease. Our findings support the concept that deregulation of the beta-catenin signaling pathway is the major oncogenic consequence of adenomatous polyposis coli mutations in intestinal neoplasia. PMID- 10463574 TI - Mutational activation of the beta-catenin proto-oncogene is a common event in the development of Wilms' tumors. AB - Activation of beta-catenin-mediated transcription is the nuclear end point of organ-specific Wnt signaling. In the developing kidney, Wnt-4, a secreted glycoprotein, acts as an autoinducer of the mesenchymal to epithelial transition that underlies normal nephron development. Dysregulation of this epithelial transformation process may lead to Wilms' tumors (WTs). In this study, we investigated the potential role of the beta-catenin proto-oncogene, a candidate downstream target molecule of Wnt-4 signaling, in the development of WTs. In 6 of 40 tumors (15%), mutation analysis revealed heterozygous missense mutations or small deletions that result in the loss of important regulatory phosphorylation sites within the beta-catenin protein. These findings indicate that activating beta-catenin mutations may play a significant role in the development of WTs and establish a direct link between Wilms' tumorigenesis and the Wnt signal transduction pathway governing normal kidney development. PMID- 10463575 TI - Common nonsense mutations in RAD52. AB - RAD51, RAD52, and RAD54 encode proteins that are critical to the repair of double strand DNA breaks by homologous recombination. The physical interactions among the products of RAD51, BRCA1, and BRCA2 have suggested that the BRCA1 and BRCA2 breast cancer susceptibility genes may function, at least in part, in this DNA damage repair pathway. Given the observation that different genes within a common functional pathway may be targeted by mutations in human cancers, we analyzed RAD51, RAD52, and RAD54 for the presence of germ-line mutations in 100 cases with early-onset breast cancer and for somatic mutations in 15 human breast cancer cell lines. Two premature stop codons, Ser346ter and Tyr415ter, were identified in germ-line RAD52 alleles from 5% of early-onset breast cancer cases. Together, these two heterozygous mutations were also found in 8% of a healthy control population, indicating that they do not confer an increased risk for breast cancer. A rare germ-line missense mutation was identified in RAD54, whereas no sequence variants were found in RAD51. None of the three RAD genes demonstrated somatic mutations in breast cancer cell lines. We conclude that, despite their potential functional association with the BRCA gene products, RAD51, RAD52, and RAD54 are not themselves targeted by mutations in human breast cancer. The presence of common nonsense mutations in RAD52 within the population may have significance for other conditions associated with potential alterations in DNA damage repair pathways. PMID- 10463576 TI - Mosaic allelic insulin-like growth factor 2 expression patterns reveal a link between Wilms' tumorigenesis and epigenetic heterogeneity. AB - Numerous observations link the loss of imprinting of insulin-like growth factor 2 (IGF2) and an overdosage of this growth factor gene with cancer, in general, and with Wilms' tumorigenesis, in particular. It is not known, however, if loss of imprinting correlates with specific stages of neoplasia or if allelic expression patterns vary within the tumor. By applying an allele-specific in situ hybridization technique to formalin-fixed thin sections, we show that the parental IGF2 alleles can be differentially expressed, not only in Wilms' tumors, but also in nephrogenic rests (which represent premalignant lesions) of Wilms' tumor patients. Moreover, a subpopulation of mesenchymal cells, which surrounds tumor nodules, expresses IGF2 biallelically irrespective of the imprinted state of IGF2 within the tumor. These data show that Wilms' tumorigenesis involves epigenetic heterogeneity as visualized by variable allelic IGF2 expression patterns. PMID- 10463577 TI - Mutagenic activation of environmental carcinogens by microsomes of gastric mucosa with intestinal metaplasia. AB - Coexpression of cytochrome P450 monooxygenases (CYPs) and reductase was found in human gastric mucosa with intestinal metaplasia. Immunohistochemistry showed reactivity to P450 reductase in metaplastic epithelial cells and in pyloric gland cells in glands showing intestinal metaplasia. These cells exhibit NADPH diaphorase activity. Reverse transcription-PCR analysis and Western blotting showed that CYP1A1 and CYP1A2 were expressed in specimens with intestinal metaplasia. Tissue distribution of CYP1A1 coincided with that of P450 reductase. However, immunoreactivity to CYP1A2 protein was localized only in the pyloric gland cells near the intestinal metaplastic gland. Salmonella typhimurium mutagen assay definitively revealed that microsomes prepared from gastric mucosa with intestinal metaplasia, in particular in the pyloric gland, functionally activated benzo(a)pyrene and 2-amino-3-methylimidazo [4,5-f]quinoline. These results indicate that carcinogen activation by CYP enzymes expressed in the gastric mucosa may contribute to carcinogenesis of the stomach. PMID- 10463578 TI - Quantitative analysis of aberrant p16 methylation using real-time quantitative methylation-specific polymerase chain reaction. AB - We have developed a quantitative method for methylation analysis of the p16 gene based on real-time methylation-specific PCR (MSP). Real-time MSP is sensitive enough to detect down to 10 genome equivalents of the methylated p16 sequence. Application of real-time MSP to DNA from tumor-derived cell lines revealed complete concordance with conventional MSP analysis. Quantitative data generated by real-time MSP were expressed as the methylation index, which was defined as the percentage of bisulfite-converted DNA that consisted of methylated target sequences. The methylation index was shown to be inversely correlated with p16 gene transcription during demethylation treatment of cell lines with 5-aza-2' deoxycytidine. The application of real-time MSP to bone marrow aspirates from patients with multiple myeloma revealed complete concordance with conventional MSP analysis. Real-time quantitative MSP may have applications in elucidating diverse biological processes involving DNA methylation and may become a valuable diagnostic tool for detecting tumor-associated epigenetic changes in cancer patients. PMID- 10463579 TI - Different frequencies and patterns of beta-catenin mutations in hepatocellular carcinomas induced by N-nitrosodiethylamine and a choline-deficient L-amino acid defined diet in rats. AB - To allow a study of beta-catenin mutations in hepatocellular carcinomas (HCCs) induced by exogenous and endogenous carcinogens, we induced tumors in male Fischer 344 rats with N-nitrosodiethylamine and a choline-deficient L-amino acid defined diet. Administration of the former was followed by partial hepatectomy with colchicine to induce cell cycle disturbance and a selection pressure regimen (K. Ohashi et al., Cancer Res., 56: 3474-3479, 1996; M. Tsutsumi et al., Jpn. J. Cancer Res., 87: 5-9, 1996). HCCs were obtained after 42 weeks. With continuous choline-deficient L-amino acid-defined feeding, tumors were sampled after 75 weeks. Total RNA was extracted from individual lesions and mutations in the glycogen synthase kinase-3beta phosphorylation consensus motif of beta-catenin were investigated by reverse transcriptase-PCR-single-strand conformation polymorphism analysis followed by nucleotide sequencing. Changes were detected in 5 of 11 HCCs induced by the exogenous carcinogen. The observed shifts of C:G- >G:C or C:G-->A:T at codon 33 and G:C-->T:A transversions at codon 34 were associated with beta-catenin protein accumulation and confirmed by Western blot analysis. Only 2 of 15 HCCs induced in the endogenous carcinogenesis regimen demonstrated mutations, those being transitions of C:G-->T:A at codon 41 without amino acid alteration. These results suggest that different genetic pathways underlie exogenous and endogenous liver carcinogenesis in rats. PMID- 10463580 TI - Cytochrome P450c17alpha gene (CYP17) polymorphism predicts use of hormone replacement therapy. AB - We investigated whether a polymorphism in the cytochrome P450c17alpha gene (CYP17), which is associated with higher endogenous hormone levels, influences the use of hormone replacement therapy (HRT). The study included 749 postmenopausal women ages 44-75 years at baseline randomly selected from a larger multiethnic cohort. African-American, Japanese, Latina, and white women were included in the study. Women who carry the CYP17 A2/A2 genotype were about half as likely as women with the A1/A1 genotype to be current HRT users (odds ratio = 0.52; 95% confidence interval, 0.31-0.86). This association was present in all four racial/ethnic groups and for women above and below the median weight of 150 pounds. These findings suggest that the actual risk of breast cancer associated with HRT use may be higher than previously reported. PMID- 10463581 TI - Hammerhead ribozyme-mediated inactivation of mutant RET in medullary thyroid carcinoma. AB - Activating mutations of the RET proto-oncogene cause hereditary medullary thyroid carcinoma. To examine whether selective inactivation of mutant RET could prevent transformation, a hammerhead ribozyme was designed to cleave RET mRNA containing a transforming mutation of codon 634 TGC --> TAC (Cys634Tyr). In vitro RNA cleavage assay demonstrated that the ribozyme selectively cleaved RET RNA with a Cys634Tyr but not Cys634Arg or the normal sequence. Expression of ribozyme in NIH/3T3 cells prevented RET-mediated colony formation in soft agar. This inhibition required catalytically active ribozyme and was specific for the TAC mutation. Therefore, ribozymes designed to selectively target mutant RET RNA may provide an effective therapeutic in the treatment of this syndrome. PMID- 10463582 TI - Reciprocal positive regulation of hypoxia-inducible factor 1alpha and insulin like growth factor 2. AB - Hypoxia-inducible factor 1 (HIF-1) activates transcription of genes encoding glucose transporters, glycolytic enzymes, vascular endothelial growth factor, and other proteins involved in O2 homeostasis and tumor progression. The expression and transcriptional activity of the HIF-1alpha subunit is regulated by the cellular O2 concentration. We demonstrate that insulin, insulin-like growth factor (IGF)-1, and IGF-2 induce expression of HIF-1alpha, which is required for expression of genes encoding IGF-2, IGF-binding protein (IGFBP)-2 and IGFBP-3. These data provide a novel mechanism by which HIF-1alpha overexpression may occur in tumor cells and contribute to an autocrine growth factor loop. PMID- 10463583 TI - Antitumor activity of 13,14-dihydro-15-deoxy-delta7-prostaglandin-A1-methyl ester integrated into lipid microspheres against human ovarian carcinoma cells resistant to cisplatin in vivo. AB - One of the delta7-prostaglandin A1 derivatives with unique antitumor activities, 13,14-dihydro-15-deoxy-delta7-prostaglandin-A1-methyl ester, was integrated into lipid microspheres (Lipo-TEI9826) and examined for its antitumor effect in vitro and in vivo. The in vitro relative resistance of human ovarian cancer, A2780CP, to cisplatin (CDDP) and Lipo-TEI9826 was 27.3 and 2.0, respectively, compared with A2780, the parent cell line of A2780CP. In in vivo experiments, when A2780CP and the parent cell line A2780 were inoculated into nude mice, A2780CP grew two times more rapidly than did A2780. The growth of A2780CP tumor was not suppressed by CDDP, whereas that of the A2780 tumor was significantly suppressed. Nevertheless, the growth of both the A2780 and the A2780CP inoculated tumors was significantly inhibited by treatment with Lipo-TEI9826 at any time after the initial treatment, compared with the lipid microspheres only. These results show that Lipo-TEI9826 may be an effective antitumor agent and capable of overcoming CDDP resistance. PMID- 10463584 TI - An insulin-like growth factor-mediated, phosphatidylinositol 3' kinase independent survival signaling pathway in beta tumor cells. AB - Hyperproliferation of tumor cells usually coincides with increased tumor cell apoptosis. To overcome apoptosis, tumor cells frequently induce the expression of growth factors that mediate cell survival. In nontransformed cells, including fibroblasts and neurons, survival factor-mediated signal transduction involves the activation of phosphatidylinositol 3' kinase (PI-3K) and protein kinase B/c Akt (PKB). Here we demonstrate that tumor cell lines derived from a transgenic mouse model of pancreatic beta cell carcinogenesis use insulin-like growth factors to repress apoptosis independently of PI-3K and PKB. The results indicate that tumor cells can use additional survival signal transduction pathways. PMID- 10463585 TI - Identification of a novel complex between human kallikrein 2 and protease inhibitor-6 in prostate cancer tissue. AB - Human kallikrein (hK) 2 is an arginine-selective serine protease expressed predominantly in the prostate that has an 80% sequence identity with prostate specific antigen. Expression of hK2 is elevated in the tumor epithelium compared to benign prostate tissue. We have purified, sequenced, and identified a novel hK2 complex in prostate tissue consisting of hK2 and a serine protease inhibitor known as protease inhibitor-6 (PI-6). This 64-kDa SDS-PAGE stable complex is elevated in the tumor and is approximately 10% of total hK2. No comparable complex of prostate-specific antigen was detected. PI-6, also known as cytoplasmic antiprotease, has been characterized as an intracellular inhibitor of trypsin and chymotrypsin-like proteases, which has high homology to plasminogen activator inhibitor 1 and 2. The physiological role of PI-6 in the prostate and its relationship to hK2 and prostate cancer are under investigation. PMID- 10463586 TI - Decreases in Ikaros activity correlate with blast crisis in patients with chronic myelogenous leukemia. AB - Gene targeting studies in mice have shown that the lack of Ikaros activity leads to T-cell hyperproliferation and T-cell neoplasia, establishing the Ikaros gene as a tumor suppressor gene in mice. This prompted us to investigate whether mutations in Ikaros play a role in human hematological malignancies. Reverse transcription-PCR was used to determine the relative expression levels of Ikaros isoforms in a panel of human leukemia/lymphoma cell lines and human bone marrow samples from patients with hematological malignancies. Among the cell lines examined, only BV-173, which was derived from a chronic myelogenous leukemia (CML) patient in lymphoid blast crisis, overexpressed the dominant-negative isoform, Ik-6. In 9 of 17 samples of patients in blast crisis of CML, Ikaros activity had been reduced either by drastically reducing mRNA expression (4 of 17) or by overexpressing the dominant-negative isoform Ik-6 (5 of 17). Significantly, expression of Ikaros isoforms seemed normal in chronic phase CML patients and patients with other hematological malignancies. In some cases, overexpression of the dominant-negative Ik-6 protein was confirmed by Western blot analysis, and Southern blot analysis indicated that decreases in Ikaros activity correlated with a mutation in the Ikaros locus. In summary, these findings suggest that a reduction of Ikaros activity may be an important step in the development of blast crisis in CML and provide further evidence that mutations that alter Ikaros expression may contribute to human hematological malignancies. PMID- 10463587 TI - Effects of geldanamycin on signaling through activator-protein 1 in hypoxic HT29 human colon adenocarcinoma cells. AB - One of the characteristic responses of HT29 human colon adenocarcinoma cells to hypoxic stress is the induction of c-jun expression and binding to the activator protein 1 (AP-1) element. To study the mechanism of c-jun activation during hypoxia, inhibitors of signaling pathways leading to the activation of AP-1 transcription factor were used. One of them, the benzoquinone ansamycin geldanamycin (GA) Mr-90,000 heat-shock protein (hsp90)-binding antibiotic, is known to disrupt signaling pathways by inducing destabilization of the enzyme complexes and degradation of signaling intermediates involving the proteasome. In our experiments, GA inhibited both basal and hypoxia-induced c-jun expression (IC50 = 75 nM). GA also abolished the hypoxia-induced increase in c-Jun NH2 terminal kinase (JNK1) catalytic activity and demonstrated an inhibitory effect on stress-activated protein kinase/ERK kinase-1 (SEK1); other participants in the mitogen-activated protein kinase and p38 signal transduction pathways were not affected to the same degree. GA treatment led to a decrease in the nuclear content of c-Jun but not that of c-Fos or of activating transcription factor 2. Functional consequences of these effects were suggested by the inhibition of AP-1 binding in hypoxic HT29 cells in the presence of GA. Pretreatment with the proteasome inhibitor lactacystin before the addition of GA resulted in the elevation of overall c-jun level, but it was unable to restore the hypoxia induced c-jun expression. Our results demonstrate that GA acts as a highly potent inhibitor of hypoxia-induced c-jun expression, affecting the activation of JNK and of the AP-1 transcription factor. However, the effect of GA cannot be attributed solely to the inhibition of signaling through JNK, and additional mechanisms remain to be identified. PMID- 10463588 TI - Two regions of deletion in 9p23-24 in sporadic breast cancer. AB - Allelic deletions of 9p including band 21-22 are common in various types of human carcinomas including breast cancer. Our previous cytogenetic studies had identified constitutional chromosomal changes in 9p23-24 in patients of a male breast-cancer family and 9p23-24 alterations in a cell line established from a sporadic female breast cancer. To find out whether this genomic region is involved more frequently in alterations in sporadic breast cancers, we have surveyed 80 microdissected tumor samples for both loss of heterozygosity (LOH) and homozygous deletion at 22 microsatellite loci spanning 9p22 to 9p24 using fluorescent multiplex PCR. LOH at one or more loci was observed in 32 (40%) of these tumors. Homozygous deletion was detected in four cases. Eleven tumors had LOH at all of the informative loci analyzed, whereas 21 tumors showed partial terminal or interstitial allelic loss of 9p. Deletion mapping identified two common regions of deletion: (a) 4 cM including D9S281 to D9S286; and (b) 1 cM including D9S1808 to D9S268. PMID- 10463589 TI - P-glycoprotein and cytochrome P-450 3A inhibition: dissociation of inhibitory potencies. AB - Many P-glycoprotein (P-gp) inhibitors studied in vitro and in vivo are also known or suspected to be substrates and/or inhibitors of cytochrome P-450 3A (CYP3A). Such overlap raises the question of whether CYP3A inhibition is an intrinsic characteristic of P-gp inhibitors, a matter of concern in the development and rational use of such agents. Thus, the purpose of the present study was to determine whether the ability to inhibit P-gp and CYP3A is, in fact, linked and whether specific P-gp inhibitors with limited ability to inhibit CYP3A can be identified. Therefore, the potency of a series of 14 P-gp inhibitors was assessed by measuring their inhibition of the transepithelial flux across Caco-2 cells of digoxin, a prototypical P-gp substrate. CYP3A inhibition was determined from the impairment of nifedipine oxidation by human liver microsomes. Determination of the apparent Ki values for CYP3A inhibition and the IC50s for P-gp and CYP3A inhibition allowed comparison of the relative inhibitory potency of the compounds on the two proteins' function. The IC50s for P-gp inhibition ranged from 0.04 to 3.8 microM. All compounds inhibited CYP3A with apparent Ki values of between 0.3 and 76 microM and IC50s between 1.5 and 50 microM. However, no correlation was found between the extent of P-gp inhibition and CYP3A inhibition, and the ratio of the IC50 for CYP3A inhibition to the IC50 for P-gp inhibition varied from 1.1 to 125. These results demonstrate that, although many P-gp inhibitors are potent inhibitors of CYP3A, a varying degree of selectivity is present. The development and use of P-gp inhibitors with minimal or absent CYP3A inhibitory effects should decrease the impact of drug interactions on the therapeutic use of such compounds. PMID- 10463590 TI - Unusually high expression of N-acetylglucosaminyltransferase-IVa in human choriocarcinoma cell lines: a possible enzymatic basis of the formation of abnormal biantennary sugar chain. AB - Structural analysis of the sugar chains of human chorionic gonadotropin (hCG) has revealed that abnormal biantennary structures appear specifically on hCG in the urine of choriocarcinoma patients. However, the enzymatic and molecular mechanisms of the biosynthesis of abnormal biantennary sugar chains have not yet been elucidated. In this report, the enzyme activities and the expression levels of mRNAs of N-acetylglucosaminyltransferases (GnT)-I to -V, beta-1,4 galactosyltransferase, and alpha-mannosidase II in normal human placentae and three human choriocarcinoma cell lines were investigated. GnT-IV activities in choriocarcinoma cell lines were increased from 16- to 66-fold and GnT-III activity was increased from 15- to 25-fold as compared with those in human placentae, whereas other enzyme activities were not increased significantly. The mRNA expression levels generally correlated with their enzyme activities. Among the two GnT-IV genes found in human tissues only GnT-IVa gene was strongly expressed in the cancer cells: from three to seven times as much as in the normal tissue, whereas that of GnT-IVb remained constant. On the basis of these results, we proposed that ectopic expression of GnT-IVa gene should occur along with the malignancy of trophoblastic tissues, and that the increased GnT-IV activity should be the main cause of the formation of abnormal biantennary sugar chains in choriocarcinoma. A possible enzymatic basis of the biosynthesis of abnormal biantennary sugar chains is discussed. PMID- 10463591 TI - Growth factor signal transduction immediately after two-thirds partial hepatectomy in the rat. AB - Liver regeneration after partial hepatectomy (PHx) of the liver serves as a model for studying normal growth factor signals that become aberrant in cancer. Growth factor signals that may play a role in initiating the proliferation of hepatocytes after 70% PHx in the rat were investigated immediately after surgical resection of the liver. Presumptive activity was evaluated by determining the tyrosine phosphorylation state of receptors for epidermal growth factor (EGF) and hepatocyte growth factor (HGF) in the liver after PHx and after sham operation as a control. Under these conditions, it was determined that the EGF receptor was constitutively phosphorylated. EGF receptor tyrosine phosphorylation, however, was increased over basal levels by 60 min after resection. The HGF receptor, c Met, was minimally phosphorylated in control livers, but a biphasic increase in phosphorylation was observed at 1-5 min after PHx and 60 min postsurgery. A slight increase in c-Met phosphorylation was observed in the sham-operated livers, but the signal was significantly less when compared with that in resected livers. Furthermore, 1 min after PHx, but not sham operation, urokinase-type plasminogen activator (u-PA) and u-PA receptor were observed in the immunoprecipitates of c-Met. Signaling downstream of growth factor receptor activation was also examined. There were no discernible phosphorylation changes in focal adhesion kinase during the early events after surgery in PHx; however, a rapid and sustained increase in the tyrosine phosphorylation of paxillin beginning 1 min after PHx, and a gradual increase in the phosphorylation beginning 5 min postsham operation, were observed. Changes in the activated state of the small GTP-binding protein Rho A and its associated proteins were seen but only after 3 h after PHx. The results indicate that HGF-related signal transduction cascades, which contribute to hepatocyte proliferation, are initiated within one min after PHx. PMID- 10463592 TI - The pro-apoptotic drug camptothecin stimulates phospholipase D activity and diacylglycerol production in the nucleus of HL-60 human promyelocytic leukemia cells. AB - It has recently been reported (T. Shimizu et al., J. Biol. Chem., 273: 8669-8674, 1998) that the pro-apoptotic drug, camptothecin, an inhibitor of topoisomerase I, induces a protein kinase C-alpha-mediated phosphorylation of lamin B in HL-60 cells, which precedes both degradation of lamin B and fragmentation of DNA. In this paper, we report that, in HL-60 cells exposed to camptothecin, there is a rapid and sustained increase of nuclear protein kinase C-alpha activity that is due to an increase in the amount of protein kinase C-alpha present in the nucleus. The enhancement of nuclear kinase C activity is preceded by an increase in the mass of nuclear diacylglycerol. As demonstrated by its sensitivity to propranolol, the nuclear diacylglycerol mass increase is due to the activation of a phospholipase D. Indeed, inhibitors of neither phosphatidylcholine-specific phospholipase C nor phosphoinositide-specific phospholipase C blocked the rise in nuclear diacylglycerol. In vitro assays also demonstrated the activation of a nuclear phospholipase D, but not of a phosphoinositide-specific phospholipase C, after treatment with camptothecin. Propranolol was also able to block the rise in nuclear protein kinase C-alpha activity, thus suggesting that the increase in diacylglycerol mass is important for the activation of the kinase at the nuclear level. Moreover, propranolol was capable of drastically reducing the number of HL 60 cells that underwent apoptosis after treatment with camptothecin. Our results show the activation during apoptosis of a phospholipase D-mediated signaling pathway operating at the nuclear level. This pathway may represent an attractive therapeutic target for the modulation of apoptotic events in human disease. PMID- 10463593 TI - Efficient nucleotide excision repair of cisplatin, oxaliplatin, and Bis-aceto ammine-dichloro-cyclohexylamine-platinum(IV) (JM216) platinum intrastrand DNA diadducts. AB - Tumors exhibit a spectrum of cellular responses to chemotherapy ranging from extreme sensitivity to resistance, either intrinsic or acquired. These variable responses are both patient and tumor specific. For platinum DNA-damaging agents, drug resistance depends on the carrier ligand of the platinum complex and is due to a combination of mechanisms including DNA repair. Nucleotide excision repair is the only known mechanism by which bulky adducts, including those generated by platinum chemotherapeutic agents, are removed from DNA in human cells. In this report, we show that the types of DNA lesions generated by three platinum drugs, cisplatin, oxaliplatin, and (Bis-aceto-ammine-dichloro-cyclohexylamine platinum(IV) (JM216), are repaired in vitro with similar kinetics by the mammalian nucleotide excision repair pathway. PMID- 10463594 TI - Delayed apoptotic responses associated with radiation-induced neoplastic transformation of human hybrid cells. AB - HeLa X human skin fibroblast hybrid cells have been developed into a model for radiation-induced neoplastic transformation of human cells. Previous studies indicate that the appearance of neoplastically transformed foci in this system is delayed for several population doublings after irradiation and appears to involve the loss of putative tumor suppressor loci on fibroblast chromosomes 11 and 14. We now show that after treatment with 7 Gy of X-rays, transformed foci initiation correlates with delayed apoptosis initiated in the progeny of the irradiated cells after 10-12 cell divisions and with reduced plating efficiency (delayed death). The cells develop classic apoptotic morphology, positive terminal deoxynucleotidyl transferase-mediated nick end labeling and phosphatidylserine (annexin V) staining, and cleavage of poly(ADP-ribose) polymerase. In addition, a delayed induction of the p53 protein and the proapoptotic Bax protein is evident over a week after radiation exposure. We propose that a delayed build-up of mitosis-dependent genomic DNA damage or a loss of genetic material over time (10 12 cell divisions postirradiation) has two relevant outcomes: (a) cell death due to the delayed induction of a p53-dependent apoptosis; and (b) neoplastic transformation of a minor subset of survivors that has lost fibroblast chromosomes 11 and 14 (tumor suppressor loci for this system) and has either evaded apoptosis or not acquired enough genetic damage to induce apoptosis. It is postulated that both phenomena result from X-ray-induced, translesion-mediated genomic instability. PMID- 10463595 TI - The protein kinase C inhibitor CGP41251 suppresses cytokine release and extracellular signal-regulated kinase 2 expression in cancer patients. AB - Components of cell signaling pathways provide important targets for anticancer drugs. Protein kinase C (PKC) is a serine/threonine-specific kinase that regulates cell growth and differentiation. It is also implicated in tumor promotion. The staurosporine analogue CGP41251 is a PKC inhibitor, and it is currently in a Phase I clinical trial for treatment of advanced cancer. However, it is difficult to define its biological activity. We have used two approaches to measure the in vivo biological response to CGP41251: (a) sequential whole blood samples were taken from 27 patients before and during treatment and incubated with mitogen (PHA), and cytokine [tumor necrosis factor (TNF)-alpha and interleukin (IL)-6] release was measured ex vivo; and (b) peripheral blood lymphocytes were isolated from seven of these patients, and the levels of extracellular signal-regulated kinase 2 were measured by Western blotting. Response to PHA was significantly lowered during treatment (P < 0.001 for TNF alpha production; P < 0.03 for IL-6). This was most evident at 7 and 28 days after the start of treatment in patients receiving higher doses (150-300 mg/day; P = 0.002 and P = 0.02, respectively, for TNF-alpha and P = 0.001 and P = 0.003, respectively, for IL-6 release). Whole blood cytokine production returned to pretreatment levels after drug administration ceased. The levels of extracellular signal-regulated kinase 2 were reduced by 50-97% during treatment in all seven patients tested. These results show for the first time that a PKC inhibitor can block in vivo signaling pathways in cancer patients. The assays we describe complement toxicity studies in selecting relevant doses for Phase II trial of novel agents, particularly when biological activity occurs at doses below those that cause obvious side effects. PMID- 10463596 TI - Anhydroretinol induces oxidative stress and cell death. AB - The retro-retinoid anhydroretinol (AR), a physiological metabolite of retinol (vitamin A), induces cell death in multiple in vitro systems. AR-induced cell death is blocked by retinol and its metabolite 14-hydroxy-4,14-retro-retinol. AR has been shown also to prevent mammary cancer induced by N-methyl-N-nitrosourea in rats. We report that AR kills cells by generating reactive oxygen species. Direct measurements show that the addition of AR to lymphoblastoid cells increases the intracellular oxidative stress in a time- and dose-dependent manner. Furthermore, the amount of induced oxidative stress directly correlates with the number of dying cells. The addition of retinol, 14-hydroxy-4,14-retro retinol, or the antioxidant, alpha-tocopherol (vitamin E), decreases AR-induced oxidative stress and proportionally reduces AR-induced cell death. In contrast, pretreatment with caspase inhibitors, known to inhibit apoptosis, has no effect on AR-induced cell death. This is the first demonstration of cellular reactive oxygen species production by a natural retinoid. PMID- 10463597 TI - Indole-3-carbinol prevents cervical cancer in human papilloma virus type 16 (HPV16) transgenic mice. AB - Mice that express transgenes for human papillomavirus type 16 under a keratin 14 promoter (K14-HPV16 mice) develop cervical cancer when they are given 17beta estradiol chronically. We asked whether the antiestrogenic phytochemical indole-3 carbinol (I3C), found in cruciferous vegetables, administered at physiological doses, would prevent the cervical-vaginal cancer that is promoted in these mice by high doses of estrogen. We compared mice that were fed a control diet with those that were fed a diet supplemented with 2000 ppm I3C. In the group fed the control diet, at a dose of estradiol of 0.125 mg per 60-day release, 19 of 25 transgenic mice developed cervical-vaginal cancer within 6 months, and the remainder had dysplasia. Only 2 mice of 24 in the group fed the I3C supplemented diet developed cancer, and the remainder had dysplasia or hyperplasia. I3C reduced dysplasia in the nontransgenic mice. Similar results were obtained at a higher dose of estradiol (0.250 mg per 60-day release), and I3C helped to prevent morbidity associated with retention of fluid in the bladder that frequently occurred with the higher estradiol dose. Additionally, I3C appeared to reduce skin cancer in transgenic mice. These data indicate that I3C is a useful preventive for cervical-vaginal cancer and, possibly, other cancers with a papillomavirus component. PMID- 10463598 TI - Enhancement of antibody-directed enzyme prodrug therapy in colorectal xenografts by an antivascular agent. AB - The irregular nature of solid tumor vasculature produces a heterogeneous distribution of antibody-targeted therapies within the tumor mass, which frequently results in reduced therapeutic efficacy. We have, therefore, combined two complementary therapies: Antibody-directed Enzyme Prodrug Therapy (ADEPT), which targets tumor cells, and an agent that selectively destroys tumor vasculature. A single i.p. dose (27.5 mg/kg) of the drug 5,6-dimethylxanthenone-4 acetic acid (DMXAA), given to nude mice bearing the LS174T colorectal xenograft, destroyed all but a peripheral rim of tumor cells, without enhancing survival. The ADEPT system, in which a pretargeted enzyme activates a prodrug, consisted of the F(ab')2 fragment of anti-carcinoembryonic antigen antibody A5B7 conjugated to the bacterial enzyme carboxypeptidase G2 and the prodrug 4-[(2-chloroethyl)(2 mesyloxyethyl)amino]benzoyl-L-glutamic acid, which was given i.p. in three doses of 500 mg/kg at 72, 84, and 96 h post-conjugate administration (25 units of carboxypeptidase G2). The antibody-enzyme conjugate could be selectively retained at approximately twice the control levels by administration of the antivascular agent at the time of optimal conjugate localization within the tumor (20 h post conjugate administration), as demonstrated by gamma counting, phosphor plate image analysis, and active enzyme measurement. This resulted in significantly enhanced tumor growth inhibition in groups of six mice, compared to conventional ADEPT therapy, with no concomitant increase in systemic toxicity. In a separate experiment, aimed at trapping the prodrug within the tumor, a 16-fold increase over control values was produced (means, 44.8 versus 2.8 microg/g tumor) when DMXAA was given 4 h prior to 4-[(2-chloroethyl)(2-mesyloxyethyl)amino]benzoyl-L glutamic acid. The therapeutic window was small, with no significant enhancement of prodrug retention when DMXAA was given at either earlier or later time points. This correlated with the time of vascular shut-down induced by the antivascular agent. We are currently investigating whether it is more advantageous to trap increased levels of conjugate or prodrug within the tumor for maximal enhancement of conventional ADEPT. These studies demonstrate that combined use of antibody directed and antivascular therapies can significantly benefit the therapeutic outcome of either strategy alone. PMID- 10463599 TI - FJ5002: a potent telomerase inhibitor identified by exploiting the disease oriented screening program with COMPARE analysis. AB - To facilitate the search for candidate telomerase inhibitors, we exploited the database of the disease-oriented screening program (DOS) available in our facility by using COMPARE analysis. In primary and arbitrary screening, we were able to identify the alkaloid berberine as a moderate inhibitor with 50% inhibition at approximately 35 microM. Using this alkaloid as a seed compound in COMPARE resulted in the identification of other berberine-like compounds and mitochondria-accumulating agents as highly related to berberine. Among these compounds, MKT077, a rhodacyanine derivative currently under Phase I clinical trials, showed a potent inhibitory effect with 50% inhibition at approximately 5 microM. With MKT077 as an upgraded seed for a new round of COMPARE analysis, we identified rhodacyanine FJ5002, a close derivative of MKT077, as the most potent telomerase inhibitor with 50% inhibition at approximately 2 microM. Long-term cultivation of U937, a human leukemia cell line, with subacute concentrations of FJ5002 resulted in population-doubling dependent changes characterized by progressive telomere erosion (from approximately 10 to approximately 4.0 kb), increased chromosome abnormalities, and senescence/crisis-like features. These results indicated that FJ5002 is a genuine and effective antitelomerase agent. PMID- 10463600 TI - Ku autoantigen affects the susceptibility to anticancer drugs. AB - The Ku70/80 autoantigens (Ku) are the DNA-binding components of a DNA-dependent protein kinase (PK) involved in DNA double strand breaks repairing a V(D)J recombination. Because apoptosis is associated with DNA fragmentation and, consequently, creation of double strand breaks, and a variety of DNA-damaging drugs kill tumor cells by apoptosis, we tested the impact of Ku deficiency on the sensitivity of anticancer drugs. Ku-null mutant cell lines Ku70-/- and Ku80-/- were highly sensitive to anticancer drugs, compared with their wild-type cells. Ku-deficient cells were more sensitive to bleomycin-induced DNA fragmentation and exhibited a higher level of c-jun NH2-kinase/stress-activated PK activity than wild-type cells, whereas R7080-6 cells overexpressing both human Ku70 and Ku80 were resistant to bleomycin-induced apoptosis and exhibited a lower level of c jun NH2-kinase/stress-activated PK activity. The Ku-protein level and Ku DNA binding activity were decreased after treatment with bleomycin, adriamycin, or vincristine, and the decreases were blocked by the treatment of z-DEVD-fmk, a specific inhibitor of caspase-3, suggesting that loss of Ku DNA binding is, in part, due to a caspase-mediated decrease in Ku protein levels. By contrast, HSF1 DNA-binding activity was increased by the treatment of these anticancer drugs and, subsequently, mitochondrial heat shock protein HSP75 was specifically induced. Our data suggest that Ku can affect the susceptibility to anticancer drug-induced apoptosis. PMID- 10463601 TI - Pharmacokinetics and antitumor activity of a bivalent disulfide-stabilized Fv immunotoxin with improved antigen binding to erbB2. AB - We have generated a stable bivalent Fv molecule [(dsFv)2] of the anti-erbB2 monoclonal antibody e23 in which the V(H) and V(L) domains of the Fv are linked to each other by a disulfide bond and the two Fvs are connected by a 15-amino acid linker (T. K. Bera et al., J. Mol. Biol., 281: 475-483, 1998). The e23 (dsFv)2 molecule is linked to a truncated form of Pseudomonas exotoxin (PE38) to generate a bivalent disulfide-stabilized immunotoxin e23 (dsFv)2-PE38. Compared to the monovalent immunotoxin, the (dsFv)2 immunotoxin showed greatly increased cytotoxicity to four cancer cell lines expressing low levels of erbB2 but not to four other cell lines with high erbB2 expression. e23 (dsFv)2-PE38 was administered i.v. to mice, and its half-life was determined. The t(1/2)alpha and t(1/2)beta were 20 and 325 min, respectively, whereas the corresponding values for the monovalent dsFv immunotoxin were shorter, 6 and 52 min. The antitumor activities of the monovalent and bivalent immunotoxin were compared using mice bearing A431 tumors. Despite the fact that e23 (dsFv)2-PE38 was 13-fold more active than e23 dsFv-PE38 on A431 cells in cell culture, its antitumor activity in mice was <2-fold that of the monovalent immunotoxin. These data show that a large increase in avidity does not always lead to an increase in cytotoxic activity. Furthermore, in one of the cases in which cytotoxic activity in vitro was greatly enhanced, there was only a small increase in antitumor activity. PMID- 10463602 TI - 5,6-trans-16-ene-vitamin D3: a new class of potent inhibitors of proliferation of prostate, breast, and myeloid leukemic cells. AB - The 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] is the physiologically active form of vitamin D3 that inhibits proliferation and induces differentiation of a variety of malignant cells. We evaluated a newly synthesized vitamin D3 analogue [1,25(OH)2-16-ene-5,6-trans-D3 (Ro 25-4020)] that has a novel 5,6-trans motif. Dose-response studies showed that 1,25(OH)2-16-ene-5,6-trans-D3 had 10-100-fold greater antiproliferative activities than 1,25(OH)2D3 when measuring clonal growth of breast (MCF-7) and prostate (LNCaP) cancer cell lines as well as a myeloid leukemia cell line (HL-60). Because the chief toxicity of vitamin D3 is hypercalcemia, we examined the calcemic activity of 1,25(OH)2-16-ene-5,6-trans-D3 in mice. Remarkably, 1,25(OH)2-16-ene-5,6-trans-D3 was at least 40-fold less calcemic as compared with 1,25(OH)2D3 and 1,25(OH)2-16-ene-D3 (Ro 24-2637). To explore the mechanism by which the 1,25(OH)2-16-ene-5,6-trans-D3 analogue mediated its antiproliferative activity, several studies were performed. Pulse exposure studies showed that a 4-day pulse exposure to 1,25(OH)2-16-ene-5,6-trans D3 (10(-7) M) in liquid culture was adequate to achieve a 40% inhibition of MCF-7 clonal growth in the absence of the analogue, suggesting that the growth inhibition mediated by 1,25(OH)2-16-ene-5,6-trans-D3 was at least in part irreversible. Cell cycle studies showed that 1,25(OH)2-16-ene-5,6-trans-D3 increased the proportion of MCF-7 cells in the G0-G1 phase and decreased those in the S phase. Furthermore, 1,25(OH)2-16-ene-5,6-trans-D3 induced an elevated expression of the cyclin-dependent kinase inhibitors, p21waf1 and p27kip1. In addition, 1,25(OH)2-16-ene-5,6-trans-D3 almost completely inhibited telomerase activity, as measured by telomeric repeat amplification protocol assay and human telomerase reverse transcriptase mRNA. For each of the growth-related parameters that were examined, the vitamin D3 analogue was more active than 1,25(OH)2D3. In contrast, 1,25(OH)2D3 was more calcemic than 1,25(OH)2-16-ene-5,6-trans-D3. In summary, 1,25(OH)2-16-ene-5,6-trans-D3, having a novel 5,6-trans motif, strongly inhibited clonal proliferation and reduced telomerase activity with low calcemic activity, suggesting further testing in in vivo cancer models. This analogue may gain a therapeutic niche for selected malignancies. PMID- 10463603 TI - Antisense Bcl-2 oligodeoxynucleotides inhibit progression to androgen independence after castration in the Shionogi tumor model. AB - Progression to androgen-independence remains the main obstacle to improving survival for patients with advanced prostate cancer. Although Bcl-2 expression in normal prostatic epithelial cells is low or absent, Bcl-2 is highly up-regulated in prostate cancer cells after androgen withdrawal and during progression to androgen-independence. Here, we test the efficacy of antisense Bcl-2 oligodeoxynucleotide (ODN) therapy administered adjuvantly after castration to delay time to androgen-independent recurrence in the androgen-dependent mouse Shionogi tumor model. Treatment of Shionogi tumor cells in vitro with antisense Bcl-2 ODN inhibited Bcl-2 expression in a dose-dependent and sequence-specific manner. Systemic administration of antisense Bcl-2 ODN in mice bearing Shionogi tumors beginning 1 day postcastration resulted in a more rapid regression of tumors and a significant delay of emergence of androgen-independent recurrent tumors. Furthermore, despite significant reduction of Bcl-2 expression in tumor tissues, antisense Bcl-2 ODN had no effect on Bcl-2 expression in normal mouse organs. These findings illustrate the potential utility of antisense Bcl-2 therapy for prostate cancer in an adjuvant setting with androgen ablation. PMID- 10463604 TI - Induction of systemic and therapeutic antitumor immunity using intratumoral injection of dendritic cells genetically modified to express interleukin 12. AB - Bone marrow-derived dendritic cells (BM-DCs) retrovirally transduced with genes encoding murine interleukin (IL)-12 stably expressed bioactive IL-12 protein at high levels. Intratumoral injection with IL-12 gene-modified BM-DCs resulted in regression of day 7 established weakly immunogenic tumors (MCA205, B16, and D122). This antitumor effect was substantially better than that of IL-12 transduced syngeneic fibroblasts or nontransduced BM-DCs. Furthermore, intratumoral injection with IL-12-transduced dendritic cells (DCs) induced specific TH1-type responses to the tumor in regional lymph nodes and spleen at levels greater than those of IL-12-transduced fibroblasts or nontransduced BM DCs. Trafficking studies confirmed that intratumorally injected IL-12-transduced DCs, but not fibroblasts, could migrate to the draining lymph node to the same extent as nontransduced BM-DCs. This strategy designed to deliver genetically modified DCs to tumor sites is associated with systemic and therapeutic antitumor immunity and is an alternative approach to those that use delivery of DCs loaded with tumor antigen. These results support the clinical application of IL-12 gene modified DCs in patients with cancer. PMID- 10463605 TI - Potent antitumor activity of MS-247, a novel DNA minor groove binder, evaluated by an in vitro and in vivo human cancer cell line panel. AB - We synthesized a novel anticancer agent MS-247 (2-[[N-[1-methyl-2-[5-[N-[4-[N,N bis(2-chloroethyl) amino] phenyl]] carbamoyl]-1H-benzimidazol-2-yl] pyrrol-4-yl] carbamoyl] ethyldimethylsulfonium di-p-toluenesulfonate) that has a netropsin like moiety and an alkylating residue in the structure. We evaluated antitumor activity of MS-247 using a human cancer cell line panel coupled with a drug sensitivity database and subsequently using human cancer xenografts. The average MS-247 concentration required for 50% growth inhibition against a panel of 39 cell lines was 0.71 microM. The COMPARE analysis revealed that the differential growth inhibition pattern of MS-247 significantly correlated with those of camptothecin analogues and anthracyclins, indicating that MS-247 and the two drug groups might have similar modes of action. MS-247 exhibited remarkable antitumor activity against various xenografts. A single i.v. injection of MS-247 significantly inhibited the growth of all 17 xenografts tested, which included lung, colon, stomach, breast, and ovarian cancers. In many cases, MS-247 was more efficacious than cisplatin, Adriamycin, 5-fluorouracil, cyclophosphamide, VP-16, and vincristine and was almost comparable with paclitaxel and CPT-11; these are the most clinically promising drugs at present. MS-247 was noticeably more effective than paclitaxel (in HCT-15) and CPT-11 (in A549, HBC-4, and SK-OV-3). The toxicity of MS-247, indicated by body weight loss, was reversible within 10 days after administration. The MS-247 mode of action showed DNA binding activity at the site where Hoechst 33342 bound, inhibited topoisomerases I and II (as expected by the COMPARE analysis) blocked the cell cycle at the G2-M phase, and induced apoptosis. These results indicate that MS-247 is a promising new anticancer drug candidate to be developed further toward clinical trials. PMID- 10463606 TI - Analysis of the cytolytic T lymphocyte response of melanoma patients to the naturally HLA-A*0201-associated tyrosinase peptide 368-376. AB - The human tyrosinase gene codes for two distinct antigens that are recognized by HLA-A*0201-restricted CTLs. For one of them, tyrosinase peptide 368-376, the sequence identified by mass spectrometry in melanoma cell eluates differs from the gene-encoded sequence as a result of posttranslational modification of amino acid residue 370 (asparagine to aspartic acid). Here, we used fluorescent tetrameric complexes ("tetramers") of HLA-A*0201 and tyrosinase peptide 368-376 (YMDGTMSQV) to characterize the CD8+ T-cell response to this antigen in lymphoid cell populations from HLA-A2 melanoma patients. Taking advantage of the presence of significant numbers of tetramer-positive CD8+ T cells in tumor-infiltrated lymph node cells from a melanoma patient, we derived polyclonal and monoclonal tyrosinase peptide 368-376-specific CTLs by tetramer-guided flow cytometric sorting. These CTLs efficiently and specifically lysed HLA-A*0201- and tyrosinase positive melanoma cells. As assessed with tyrosinase peptide variants, the fine antigen specificity of the CTLs was quite diverse at the clonal level. Flow cytometric analysis of PBMCs stained with tetramers showed that tyrosinase peptide 368-376-specific CD8+ T cells were hardly detectable in peripheral blood of melanoma patients. However, significant numbers of such cells were detected after short-term stimulation of CD8+ lymphocytes with tyrosinase peptide 368-376 in 6 of 10 HLA-A2 melanoma patients. Taken together, these findings emphasize the significant contribution of the natural tyrosinase peptide 368-376 to the antigenic specificities recognized by the tumor-reactive CTLs that may develop in HLA-A2 melanoma patients. PMID- 10463607 TI - Identification of a gene coding for a protein possessing shared tumor epitopes capable of inducing HLA-A24-restricted cytotoxic T lymphocytes in cancer patients. AB - Genes encoding tumor epitopes that are capable of inducing CTLs against adenocarcinomas and squamous cell carcinomas, two major human cancers histologically observed in various organs, have rarely been identified. Here, we report a new gene from cDNA of esophageal cancer cells that encodes a shared tumor antigen recognized by HLA-A2402-restricted and tumor-specific CTLs. The sequence of this gene is almost identical to that of the KIAA0156 gene, which has been registered in GenBank with an unknown function. This gene encodes a Mr 140,000 protein that is expressed in the nucleus of all of the malignant tumor cell lines tested and the majority of cancer tissues with various histologies, including squamous cell carcinomas, adenocarcinomas, melanomas, and leukemia cells. However, this protein was undetectable in the nucleus of any cell lines of nonmalignant cells or normal tissues, except for the testis. Furthermore, this protein was expressed in the cytosol of all of the proliferating cells, including normal cells and malignant cells, but not in normal tissues, except for the testis and fetal liver. Two peptides of this protein were recognized by HLA-A2402 restricted CTLs and were able to induce HLA-A24-restricted and tumor-specific CTLs from peripheral blood mononuclear cells of most of HLA-A24+ cancer patients tested, but not from peripheral blood mononuclear cells of any healthy donors. These peptides may be useful in specific immunotherapy for HLA-A24+ cancer patients with various histological types. PMID- 10463608 TI - Antitumor effects of interferon-omega: in vivo therapy of human tumor xenografts in nude mice. AB - The antitumor effect of the type I IFN, IFN-omega, was evaluated in both in vitro and in vivo studies of human cancer. For these studies, the cDNA for human IFN omega was cloned into a eukaryotic expression plasmid DNA (pDNA) driven by the cytomegalovirus promoter. Supernatants from UM449 cells transfected in vitro with IFN-omega pDNA had antiproliferative effects on 11 of 13 human tumor cell lines. For in vivo studies, nude mice were implanted s.c. with one of the following human tumors: NIH: OVCAR-3 ovarian carcinoma, A375 melanoma, or A431 epidermoid carcinoma. Direct intratumoral injection of 100 microg of a IFN-omega pDNA DMRIE/DOPE complex (1:1 DNA:DMRIE mass ratio) for 6 consecutive days resulted in a significant reduction in the tumor volume of NIH: OVCAR-3 ovarian carcinoma or A375 melanoma (P = 0.02). IFN-omega pDNA delivered by i.m. injection also had an antitumor effect. Nude mice bearing s.c. A431 epidermoid carcinoma and injected i.m. with 100 microg of IFN-omega pDNA, twice per week for 3 weeks, had a significant reduction in tumor volume (P = 0.009). These results demonstrate for the first time that IFN-omega can have in vivo antitumor effects in several models of human cancer. PMID- 10463609 TI - Identification of NKIAMRE, the human homologue to the mitogen-activated protein kinase-/cyclin-dependent kinase-related protein kinase NKIATRE, and its loss in leukemic blasts with chromosome arm 5q deletion. AB - Human acute leukemia and myelodysplasia are often associated with an interstitial deletion in chromosome arm 5q. The deleted region is hypothesized to contain tumor suppressor loci that are critical to the maintenance of normal hematopoiesis. We have identified NKIAMRE, a novel cyclin-dependent kinase related molecule that is closely related to the rat serine/threonine kinase NKIATRE. Human NKIAMRE localizes to chromosome band 5q31.1, centromeric to the interleukin 9 locus and telomeric to IFN response factor-1. NKIAMRE was deleted at both alleles in 9 of 18 leukemic samples with chromosome band 5q31 abnormalities studied by fluorescence in situ chromosomal hybridization. NKIAMRE loss may be an important determinant of dysmyelopoiesis. PMID- 10463610 TI - Structure and possible mechanisms of TEL-AML1 gene fusions in childhood acute lymphoblastic leukemia. AB - TEL-AML1 gene fusion derived by chromosomal translocation is a common acquired genetic lesion in pediatric cancer that is present in approximately 25% of B-cell precursor acute lymphoblastic leukemias, and recent evidence suggests that this recombination event may initiate leukemogenesis prenatally during fetal hemopoiesis. Analysis of the DNA sequence and structure surrounding the breakpoints may reveal clues to their formation. A long-distance inverse PCR strategy was used to amplify TEL-AML1 genomic fusion sequences from diagnostic DNA from nine patients. Breakpoints were scattered within the 14 kb of intronic DNA between exons 5 and 6 of TEL and in two putative cluster regions within AML1 intron 1. Fusion sequences exhibited characteristic signs of nonhomologous end joining, including microhomologies at the end points, and small deletions and duplications. DNA sequences near the breakpoints did not reveal any consistent characteristic signal sequences of the V(D)J recombinase, topoisomerase II consensus sites, or other sequence motifs associated with recombination. However, several translocations occurred near a repeat region of TEL that was found to be highly polymorphic. This region was cloned and found in nuclease sensitivity assays to exhibit paranemic structures, which may have contributed to DNA breakage or illegitimate recombination. The data are compatible with the possibility that TEL-AML1 translocations occur by nonhomologous recombination involving imprecise, constitutive repair processes following DNA double-strand breaks. PMID- 10463611 TI - Two novel mucin genes down-regulated in colorectal cancer identified by differential display. AB - Epithelial mucins are large, secreted and cell surface glycoproteins involved in epithelial cell protection, adhesion modulation, and signaling. Using differential display, we have identified two novel mucin cDNAs (dd34 and dd29), hereafter designated MUC11 and MUC12, respectively, that are down-regulated in colorectal cancers. Northern blots demonstrated polydisperse signals characteristic of mucin transcripts in RNA from normal colon that were absent in colorectal cancer. Both cDNAs were mapped by fluorescence in situ hybridization to chromosome band 7q22, the location of the MUC3 mucin gene, thus suggesting that there may be a cluster of mucin genes at this locus. The sequences of both differential display clones were extended by a combination of screening libraries and PCR. The 2.8-kb MUC11 cDNA composite encoded 35 serine/threonine-rich, mucin like degenerate 28 amino acid tandem repeats. The MUC12 cDNA composite encoded a putative transmembrane mucin containing two extracellular cysteine-rich, EGF-like domains, a coiled-coil region, and a mucin-like domain consisting of 28 amino acid degenerate tandem repeats. Distinct patterns of expression of MUC11, MUC12, and MUC3 mRNAs were observed in a range of normal human tissues. MUC12 mRNA was not expressed in any of six colorectal cancer cell lines examined and was down regulated or absent in 6 of 15 (40%) tumors compared with matched normal colonic tissue. In contrast, MUC11 showed a different pattern of mRNA expression, with four of these lines showing low levels and the other two lines showing relatively high levels of MUC11 transcripts. Expression of MUC11 was down-regulated in the tumors of 12 of 15 (80%) paired samples. Structural homology of MUC12 with rat, mouse, and human MUC3 and human and rat MUC4/ASGP2 indicate that there is a distinct subfamily of transmembrane mucins with conserved epidermal growth factor domains. The homology of MUC12 with epidermal growth factor-like growth factors and its down-regulation in colorectal cancers, together with known interactions between rat MUC4 and c-erbB-2 growth factor receptors, suggests that MUC12 may be involved in epithelial cell growth regulation. PMID- 10463612 TI - Nuclear factor kappaB transactivation is increased but is not involved in the proliferative effects of thioredoxin overexpression in MCF-7 breast cancer cells. AB - Thioredoxin (Trx) is a small redox-active protein that provides reducing equivalents for key cysteine residues of proteins through thiol-disulfide exchange, such as the transcription factor nuclear factor-kappaB (NF-kappaB). NF kappaB activation has been associated previously with cell growth and the inhibition of apoptosis. We have shown in earlier studies that overexpression of Trx in MCF-7 cells increases anchorage-independent growth. In this study, the activation of NF-kappaB was examined as a mechanism through which Trx overexpression might promote anchorage-independent growth. Constitutive NF-kappaB activity is elevated 4-7-fold in Trx-overexpressing cells. NF-kappaB activity was inhibited in these cells by expressing a dominant-negative mutant of the IkappaB alpha protein (IkappaB alphaM). Expression of IkappaB alphaM in Trx overexpressing cells dramatically reduced the Trx-associated increase in NF kappaB activity but did not affect anchorage-dependent or -independent growth. The results suggest that increased growth in MCF-7 cells overexpressing Trx is not mediated by increased activation of the transcription factor, NF-kappaB. Additionally, activator protein-1 (AP-1), another transcription factor associated with growth, was increased up to 10-fold in Trx-overexpressing cells. Thus, AP-1 activation might contribute to the growth-promoting effect of Trx. PMID- 10463613 TI - A lack of a functional NAD(P)H:quinone oxidoreductase allele is selectively associated with pediatric leukemias that have MLL fusions. United Kingdom Childhood Cancer Study Investigators. AB - Rearrangements and fusion of the MLL gene with various alternative partner genes occur in approximately 80% of infant leukemias and are acquired during fetal hemopoiesis in utero. Similar MLL gene recombinants also occur in topoisomerase II-inhibiting drug-induced leukemias. These data have led to the suggestion that some infant leukemia may arise via transplacental fetal exposures during pregnancy to substances that form cleavable complexes with topoisomerase II and induce illegitimate recombination of the MLL gene. A structural feature shared by many topoisomerase II-inhibiting drugs and other chemicals is the quinone moiety. We assayed, by PCR-RFLP, for a polymorphism in an enzyme that detoxifies quinones, NAD(P)H:quinone oxidoreductase (NQO1), in a series (n = 36) of infant leukemias with MLL rearrangements versus unselected cord blood controls (n = 100). MLL-rearranged leukemias were more likely to have genotypes with low NQO1 function (heterozygous CT or homozygous TT at nucleotide 609) than controls (odds ratio, 2.5; P = 0.015). In contrast, no significant allele bias was seen in other groups of pediatric leukemias with TEL-AML1 fusions (n = 50) or hyperdiploidy (n = 29). In the subset of infant leukemias that had MLL-AF4 fusion genes (n = 21), the bias increase in low or null function NQO1 genotypes was more pronounced (odds ratio, 8.12; P = 0.00013). These data support the idea of a novel causal mechanism in infant leukemia involving genotoxic exposure in utero and modulation of impact on a selective target gene by an inherited allele encoding a rate limiting step in a carcinogen detoxification pathway. PMID- 10463614 TI - A new MAGE gene with ubiquitous expression does not code for known MAGE antigens recognized by T cells. AB - A number of genes of the MAGE family have been shown to code for antigens that are recognized on many human tumors by autologous CTLs. These antigens should be strictly tumor specific because the encoding MAGE genes are not expressed in normal adult cells, except for male germ-line cells, which lack HLA expression. Here, we report that a distant relative of the previously identified MAGE genes is expressed in many, if not all, normal tissues. This gene, which was named MAGE D, is located in Xp11. Its exon-intron structure is completely different from that of the other MAGE genes. None of the 20 MAGE antigenic peptides presently known to be recognized by T lymphocytes is encoded by the new MAGE gene. It appears, therefore, that this new finding leaves intact the tumor specificity of the antigens encoded by the MAGE genes that are expressed only in tumor and germ line cells. PMID- 10463615 TI - Deficiency of connexin43 gap junctions is an independent marker for breast tumors. AB - Gap junctions are intercellular channels that are formed from members of a family of proteins, the connexins (Cxs). Gap junctions play an important role in vital functions, including the regulation of cell growth and cell differentiation. Here, we examined the expression of Cx43, a major Cx in breast tissue, in 32 surgical specimens obtained from breast cancer patients who underwent a primary surgical resection prior to chemotherapy or radiotherapy treatments. The expression of Cx43 gap junctions was compared to the levels of estrogen, progesterone, and erbB2 tyrosine kinase receptors. In addition, a panel of breast cancer cell lines and a series of normal rat mammary tissues and rat mammary tumors induced in vivo by dimethylbenz(a)anthracene were studied. We demonstrated that the lack of Cx43 gap junctions is a common feature of human mammary cancer tissues compared to nonneoplastic breast tissues surrounding primary tumors. Cx43 gap junctions were not observed in ductal carcinomas in situ, infiltrating ductal carcinomas, and infiltrating lobular carcinomas, and they seem to be independent of estrogen, progesterone, and erbB2 receptor status. In breast cancer cell lines and rodent mammary carcinoma tissues, down-regulation of Cx43 occurs at the mRNA level, suggesting a transcriptional mechanism for the decrease of Cx43 protein in breast cancer. In summary, this study provides evidence of decreased expression of Cx43 gap junctions in breast cancer at various stages of progression as well as breast cancer cell lines and raises the possibility that Cx43 may be a useful marker for detecting early oncogenesis in the breast. Because Cx43 gap junctions are lacking in breast cancer and restoration of Cx43 has been shown to reverse the malignant phenotype in vitro, pharmacological up-regulation of Cx43 may prove beneficial in cancer therapeutics. PMID- 10463616 TI - Inhibition of bladder carcinoma angiogenesis, stromal support, and tumor growth by halofuginone. AB - We have previously demonstrated that halofuginone, a widely used alkaloid coccidiostat, is a potent inhibitor of collagen alpha1(I) and matrix metalloproteinase 2 gene expression. Halofuginone also suppresses extracellular matrix deposition and cell proliferation. We investigated the effect of halofuginone on transplantable and chemically induced mouse bladder carcinoma. In both systems, oral administration of halofuginone resulted in a profound anticancerous effect, even when the treatment was initiated at advanced stages of tumor development. Although halofuginone failed to prevent proliferative preneoplastic alterations in the bladder epithelium, it inhibited further progression of the chemically induced tumor into a malignant invasive stage. Histological examination and in situ analysis of the tumor tissue revealed a marked decrease in blood vessel density and in both collagen alpha1(I) and H19 gene expression. H19 is regarded as an early marker of bladder carcinoma. The antiangiogenic effect of halofuginone was also demonstrated by inhibition of microvessel formation in vitro. We attribute the profound antitumoral effect of halofuginone to its combined inhibition of the tumor stromal support, vascularization, invasiveness, and cell proliferation. PMID- 10463617 TI - Expression level of Bcl-2 determines anti- or proapoptotic function. AB - Bcl-2 is an oncogene with antiapoptotic function. However, Bcl-2 is converted to a Bax-like death effector by caspases, suggesting that the expression of Bcl-2 may not favor the growth of cancers. We introduced the Bcl-2 gene to gliomas via adenovirus (Adv; Adv-Bcl-2) with the Adv for Fas (Adv-Fas) and the Adv for Fas ligand (Adv-FL) to evaluate the antiapoptotic function of Bcl-2. In U251 glioblastoma cells, Bcl-2 at a low level of expression repressed apoptosis induced by Adv-Fas and Adv-FL, whereas Bcl-2 at a high level of expression did not. On the other hand, Bcl-X(L) showed antiapoptotic function against Fas mediated apoptosis, irrespective of its expression level. In glioblastoma cells, induction of Bcl-2 alone at a high level induced apoptosis, whereas induction of Bcl-X(L) alone did not. As the multiplicity of infection of Adv-Bcl-2 was increased, the quantity of a cleaved product of Bcl-2 increased. Induction of caspase-inhibitory genes (CrmA and p35) inhibited apoptosis induced by Adv-Bcl-2. Induction of Bcl-2 led to alteration of the membrane potential and structure of the mitochondria. In summary, although Bcl-2 at a low level of expression was antiapoptotic, Bcl-2 at a high level of expression was proapoptotic to Fas mediated apoptosis. Overexpression of Bcl-X(L) was consistently antiapoptotic to Fas-mediated apoptosis. PMID- 10463618 TI - Augmentation of transvascular transport of macromolecules and nanoparticles in tumors using vascular endothelial growth factor. AB - The goal of this investigation was to measure changes in vascular permeability, pore cutoff size, and number of transvascular transport pathways as a function of time and in response to vascular endothelial growth factor (VEGF), placenta growth factor (PIGF-1 and PIGF-2), or basic fibroblast growth factor (bFGF). Two human and two murine tumors were implanted in the dorsal skin chamber or cranial window. Vascular permeability to BSA (approximately 7 nm in diameter) and extravasation of polyethylene glycol-stabilized long-circulating liposomes (100 400 nm) and latex microspheres (approximately 800 nm) were determined by intravital microscopy. Vascular permeability was found to be temporally heterogeneous. VEGF superfusion (100 ng/ml) significantly increased vascular permeability to albumin in normal s.c. vessels, whereas a 30-fold higher dose of VEGF (3000 ng/ml) was required to increase permeability in pial vessels, suggesting that different tissues exhibit different dose thresholds for VEGF activity. Furthermore, VEGF superfusion (1000 ng/ml) increased vascular permeability to albumin in a hypopermeable human glioma xenograft in cranial window, whereas VEGF superfusion (10-1000 ng/ml) failed to increase permeability in a variety of hyperpermeable tumors grown in dorsal skin chamber. Interestingly, low-dose VEGF treatment (10 ng/ml) doubled the maximum pore size (from 400 to 800 nm) and significantly increased the frequency of large (400 nm) pores in human colon carcinoma xenografts. PIGF-1, PIGF-2, or bFGF did not show any significant effect on permeability or pore size in tumors. These findings suggest that exogenous VEGF may be useful for augmenting the transvascular delivery of larger antineoplastic agents such as gene targeting vectors and encapsulated drug carriers (typical range, 100-300 nm) into tumors. PMID- 10463619 TI - Available volume fraction of macromolecules in the extravascular space of a fibrosarcoma: implications for drug delivery. AB - Steric exclusion of molecules in the extravascular space of tissues can be quantified by the available volume fraction (K(AV)). Despite its clinical importance, however, there is a paucity of data in the literature regarding the available volume fraction of macromolecules in the extravascular space of tumor tissues. In this study, we quantified K(AV) of inulin, BSA, and dextran molecules of Mr 10,000-2,000,000 in polymer gels and fibrosarcoma tissues. The measurement involved: (a) sectioning of gels or tumor tissues into thin slices (approximately 600 microm) using a Vibratome, (b) ex vivo incubation of the slices in solutions containing fluorescently labeled tracers, and (c) quantification of the equilibrium tracer concentrations in both slices and solutions. We found that K(AV) in gels decreased monotonically when the Mr of dextran was increased from Mr 10,000 to 2,000,000. However, K(AV) in tumor tissues was insensitive to the molecular weight of dextran in the range between Mr 10,000 and 40,000. There was a sharp decrease in K(AV) from 0.28 +/- 0.14 to 0.10 +/- 0.06 when the molecular weight was increased from Mr 40,000 to 70,000. In addition to the molecular weight dependence, K(AV) was heterogeneous in tumors, with intertumoral difference being greater than intratumoral variation. The interstitial fluid space, which was quantified by K(AV) of inulin, was 50% of the total tissue volume. These data indicate that the fraction of the extravascular volume in tumors that is accessible to large therapeutic agents is heterogeneous and depends on the size of agents. PMID- 10463620 TI - Intragenic mutation analysis of the human epidermal growth factor receptor (EGFR) gene in malignant human oral keratinocytes. AB - Alteration in epidermal growth factor receptor (EGFR) expression is frequently associated with malignant transformation of epithelial tissues, including oral mucosa. This study examines the mutations in the coding region of the human EGFR gene in normal and malignant human oral keratinocytes. To examine the intragenic mutations in the human EGFR gene, a panel of normal and malignant human oral keratinocytes were examined by a nonisotopic RNase cleavage assay. Two consistent alterations were detected. First, a polymorphism, which generates a unique BsrI restriction site, was detected at position 2073. This BsrI polymorphism was present only in malignant keratinocytes. Second, Southern blot hybridization of PCR products revealed that there is a truncated EGFR mRNA (approximately 1.5-kb) in oral squamous cell carcinoma cell lines. Similar analysis in normal cell lines revealed that this truncated EGFR transcript is also present. Immunoblotting revealed the presence of this truncated form of EGFR in all keratinocyte cell lines. These data permit us to conclude that there exists a novel truncated form of EGFR in human oral keratinocytes. Furthermore, there exists a tumor-associated BsrI polymorphic site at position 2073. The potential biological relevance of the truncated receptor and the utility of the BsrI polymorphic site for diagnostic applications are currently being explored. PMID- 10463621 TI - Cell cycle arrest and inhibition of anoikis by galectin-3 in human breast epithelial cells. AB - Galectin-3 is a member of a growing family of animal beta-galactoside-binding proteins shown to be involved in cell growth, differentiation, apoptosis resistance, and tumor progression. In the present study, we investigated whether galectin-3 can protect against apoptosis induced by the loss of cell anchorage (anoikis). Because studies suggest that cellular sensitivity to anoikis is associated with cell cycle regulation, we examined the role of galectin-3 on cell cycle regulation. Although BT549 cells (human breast epithelial cells) undergo anoikis, galectin-3-overexpressing BT549 cells respond to the loss of cell adhesion by inducing G1 arrest without detectable cell death. Galectin-3-mediated G1 arrest involves down-regulation of G1-S cyclin levels (cyclin E and cyclin A) and up-regulation of their inhibitory protein levels (p21(WAF1/CIP1) and p27KIP1). After the loss of cell anchorage, Rb protein becomes hypophosphorylated in galectin-3-overexpressing cells, as predicted from the flow cytometric analysis and immunoblot analysis of cyclins and their inhibitors. Interestingly, galectin-3 induces cyclin D1 expression (an early G1 cyclin) and its associated kinase activity in the absence of cell anchorage. On the basis of these results, we propose that galectin-3 inhibition of anoikis involves cell cycle arrest at an anoikis-insensitive point (late G1) through modulation of gene expression and activities of cell cycle regulators. The present study suggests that galectin-3 may be a critical determinant for anchorage-independent cell survival of disseminating cancer cells in the circulation during metastasis. PMID- 10463622 TI - Oral administration of policosanol inhibits in vitro copper ion-induced rat lipoprotein peroxidation. AB - Policosanol, a new cholesterol-lowering agent, is a mixture of higher aliphatic primary alcohols isolated from sugar cane (Saccharum officinarum L.) wax, which prevents the onset of espontaneously and experimentally induced atherosclerotic lesions in experimental models. Because the oxidation of low-density lipoprotein (LDL) may play a role in the pathogenesis of atherosclerosis, we investigate the effect of policosanol on copper oxidative susceptibility of rat lipoprotein fractions (VLDL + LDL). Rats fed normal diet were treated with policosanol (250 500 mg/kg/day) for up to 4 weeks. EDTA-free lipoprotein particles were oxidized in a cell-free system by the addition of copper ions, and conjugated dienes generation was monitored by changes of optical density at 234 nm. Thiobarbituric acid-reactive substances (TBARS) content and lysine-amino group reactivity were investigated. After administration, there was no change in cholesterol, triglycerides, and phospholipid content of lipoprotein fractions; however, policosanol significantly prolongs the lag time and reduces the propagation rate of diene generation. Also, policosanol reduces TBARS content and increases lysine reactivity in lipoprotein fractions treated with Cu2+. In conclusion, policosanol, in addition to its cholesterol-lowering effect, has other properties that enables it to reduce the potential of lipoprotein to undergo lipid peroxidation. Such effect can be considered of promissory value in the management of atherosclerosis. PMID- 10463623 TI - Breakfast cereal and caffeinated coffee: effects on working memory, attention, mood, and cardiovascular function. AB - This study examined the effects of breakfast cereal and caffeinated coffee on working memory, attention, mood, and cardiovascular function. One hundred and forty-four volunteers (72 male, 72 female, mean age 21 years) were assigned to one of the groups formed by combining breakfast (cereal versus no breakfast) and caffeine (caffeinated versus decaffeinated coffee) conditions. The volunteers completed a baseline session between 0800 and 0845 h. The breakfast/caffeine administration took place between 0845 and 0915 h. They then completed another test session (starting at 0945) and had a coffee break at 1045, followed by a final session starting at 1145. The results showed that those who consumed breakfast cereal had a more positive mood at the start of the test sessions, performed better on a spatial memory task, and felt calmer at the end of the test session than those in the no breakfast condition. Ingestion of caffeine had no effect on initial mood or working memory, but it did improve encoding of new information and counteracted the fatigue that developed over the test session. Caffeine increased blood pressure and pulse rate, whereas breakfast cereal consumption only had an effect on pulse. Overall, these results confirm previous findings on the effects of breakfast and caffeine, and demonstrate distinct profiles for two common examples of early-morning food and drink, breakfast cereal and caffeinated coffee. PMID- 10463624 TI - Inbred Roman high- and low-avoidance rats: differences in anxiety, novelty seeking, and shuttlebox behaviors. AB - In the present study, male inbred animals (from the 10th generation of an inbreeding program that has been carried out in parallel to that of the outbred Roman high- and low-avoidance rat lines), were compared for emotionality in different testing situations, exploratory behavior in the holeboard and two-way, active-avoidance acquisition. Compared to the inbred Roman high-avoidance (RHA I/Verh) rats, inbred Roman low-avoidance (RLA-I-Verh) rats showed higher emotionality in the open field (reduced distance travelled and number of rearings, and increased self-grooming behavior), in the elevated plus-maze test (increased number of total and open-arm entries, reduced distance travelled in the open arms, and increased self-grooming behavior), and during the habituation period in the shuttle box (decreased number of crossings, increased self-grooming behavior and defecations). Results from the hyponeophagia test were not conclusive, probably due to the test-dependent hyperactivity shown by RHA-I/Verh rats. In the holeboard apparatus, RHA-I/Verh rats explored more than RLA-I/Verh rats, especially when novel objects were located beneath the holes. Finally, RHA I/Verh animals rapidly acquired active, two-way (shuttlebox) avoidance, whereas RLA-I/Verh animals required four 50-trial sessions to achieve an assymptotic level of 30-40% avoidance. Thus, the behavioral patterns of the Roman inbred strains were very similar to those previously reported for the RHA/Verh outbred lines. Differences in locomotor activity, exploratory, and self-grooming behavior were actually greater between the inbred strains than between the outbred lines. Differences in defecation, however, although still significant, were not so pronounced as those noted previously at this laboratory with the outbred lines. PMID- 10463625 TI - Appraisal of mood and personality during hypoglycaemia in human subjects. AB - This study used the biological model of experimental hypoglycaemia to examine the effect of a manipulation in mood-state on appraisal. Controlled hypoglycaemia was achieved using the hyperinsulinaemic glucose clamp technique. Mood, appraisal, and personality traits were assessed using well validated questionnaires. Our findings 1) reaffirm the existence of multiple arousal systems in the generation of moods, 2) show that the induction of a negative mood state does lead to more negative appraisals of a life situation, and 3) show that personality traits remain stable during the experience of negative emotions and cognitions. We conclude that hypoglycaemia, by inducing a state of tense tiredness in some individuals, may lead to more negative appraisals of a life situation but does not alter people's reporting of behavioural dispositions. PMID- 10463626 TI - Anger state during acute insulin-induced hypoglycaemia. AB - This study sought to examine the effects of insulin-induced hypoglycaemia on anger state, and to describe the associations between change in the anger state and measures of anger trait and anger expression (assessed using the State-Trait Anger Expression Inventory). A hyperinsulinaemic glucose clamp was used to achieve controlled euglycaemia (5.0 mmol/L) and hypoglycaemia (2.6 mmol/L) in 18 nondiabetic subjects and 30 people with insulin-dependent diabetes mellitus (IDDM). Subjects underwent both hypoglycaemic and euglycaemic conditions, separated by 2 weeks, in a counterbalanced order. During each study condition subjects were asked to complete a questionnaire on anger state. Results at euglycaemia and hypoglycaemia were compared, and differences between the conditions were correlated with measures of anger trait and anger expression. Hypoglycaemia caused both nondiabetic and IDDM subjects to report a significant increase in feelings of anger, despite being in a nonconfrontational setting. However, there were no clear associations between an individual's change in reported anger and measures of anger trait and anger expression. No association was found between the change in anger state and the intensity of an individual's symptomatic response to hypoglycaemia. PMID- 10463627 TI - Role of aversively motivated behavior in the olfactory bulbectomy syndrome. AB - The aim of the present studies was to determine the extent to which changes in defensive behaviors could account for some of the behavioral effects of bilateral olfactory bulbectomy (OBX) in rats. Four tests of aversively-motivated behavior were conducted in bulbectomized and sham-operated rats: activity in a dimly lit or brightly lit open field, passive avoidance, foot shock-induced freezing, and defensive withdrawal. OBX reduced the duration of immobility in the open field. Bulbectomized rats exhibited less freezing in response to foot shock than sham operated rats. In the defensive-withdrawal test, bulbectomized rats made more transitions into and spent less time inside the covered enclosure than sham operated rats. The experiments thus reveal two novel paradigms for assessing the behavioral effects of OBX. The results also suggest that deficits in aversively motivated behavior, specifically defensive freezing, may comprehensively explain the putative "hyperactivity" and "passive-avoidance learning deficits" widely associated with the OBX behavioral syndrome. PMID- 10463628 TI - Colostrum mediates the development of mother preference by newborn lambs. AB - The first sucking bouts have strong rewarding properties in the establishment of a preference for the mother by newborn lambs. In this study we designed an artificial teat (Experiment 1) and a method of tube feeding (Experiment 2) to investigate the role of nonnutritive sucking and colostrum intake as reinforcers in the development of this relationship (Experiment 3). In this third experiment, lambs of the control group (n=10) had free access to the udder. In the other groups they were prevented from sucking between birth and 6 h by covering the ewe's udder while lambs received one of the following treatments: access to nonnutritive teats fitted onto the mother's udder (n=12), tube fed with colostrum (n=11), or fully deprived (n=10). When tested in a two-choice test at 24 h of age, controls and lambs receiving colostrum spent significantly more time near their mother than near the alien ewe. By contrast, deprived lambs did not. Half of the lambs having access to nonnutritive teats displayed a preference for their mother; the other half chose the alien ewe. However, most lambs that were affected by the treatments displayed a preference for their mother when a second test was performed at 48 h. In conclusion, these results demonstrate that a neonatal nasogastric infusion of colostrum has the same rewarding properties as a complete sucking bout. They also provide evidence for the first time that a relationship with the dam can be established via a nutritional signal originating from the gastrointestinal sphere. PMID- 10463629 TI - Place cells can flexibly terminate and develop their spatial firing. A new theory for their function. AB - In this study, hippocampal place cells were recorded in a behavioral paradigm previously not employed in place-cell research. Rats were exposed to the same fixed environment for as long as 8-24 h without interruption, while the firing of CA1 and CA3 place cells was monitored continuously. The first finding was that all place cells that were detected at the beginning of the recording sessions ceased to produce location-specific firing in their original firing fields within 2-12 h. This was observed despite the fact that the animals kept visiting the original firing fields, the hippocampal EEG was virtually unchanged, and the discriminated action potentials of the cells could be clearly recorded. The second finding was that some complex-spike cells that produced no spatially selective firing pattern at the beginning of the recording sessions developed location-specific discharges within 3-12 h. Thus, place cells can flexibly terminate and develop their spatial firing. even in a fixed environment and during similar behaviors, if that environment is explored continuously for a prolonged period. To explain this phenomenon, a new place-cell theory is outlined. Accordingly, the high-frequency discharges of these neurons may serve to create, under multiple extrahippocampal control and within limited periods, stable engrams for specific spatial sites in the association cortex where the cognitive map probably resides. After the creation of a stable engram, or in the absence of favorable extrahippocampal inputs, place cells may suspend their location-specific firing in the original field, and initiate the processing of another spatial site. PMID- 10463630 TI - The influence of a glucose drink on a demanding working memory task. AB - A group of young adult females, who had or had not eaten breakfast, drank either a glucose drink or a placebo. Fasting was associated with poorer performance on the Brown-Petersen task, a test of memory. A glucose drink improved the memory of those who had fasted, although it did not influence those who had eaten breakfast. In those who had fasted, the glucose drink resulted in memory comparable to those who had consumed breakfast. Those with higher levels of blood glucose upon arrival in the laboratory had better memories. In those taking a glucose drink, after an initial rise, rapidly falling levels of blood glucose were associated with better memory. PMID- 10463631 TI - Vaginocervical stimulation inhibits female-female mounting in laboratory rats. AB - The objective of this study was to examine how vaginocervical stimulation (VCS) affects female-female mounting in laboratory rats. After receiving sexual stimulation from the male, the latency for the female to mount another female was significantly longer than that of control females. In the absence of any copulatory stimulation, the latency to initiate mounting of another female was about 3 min. However, following three mounts or three intromissions by a male, the latency for the experimental female to initiate mounting increased to about 10 min, and ejaculation abolished mounting for almost 2 h. Once females began mounting, regardless of the copulatory stimulation they received prior to testing, their mounting rate (mounting frequency/2 h) did not differ from stimulus control females. Artificial VCS also inhibited female mounting and anesthetization of the vaginocervical area diminished the inhibiting effect of ejaculation. Taken together, the present results provide evidence that VCS can temporarily inhibit female mounting, and that the duration of the inhibition is related to the amount of VCS received. These data are interpreted within the perspective that female mounting behavior is not a sexual behavior and is consequently suppressed within the context of normal copulation. PMID- 10463632 TI - Caffeine-induced modifications of heart rate, temperature, and motor activity circadian rhythms in rats. AB - Even if the effects of caffeine on some physiological parameters are well known, its influence on circadian rhythmicity had not yet been investigated. This possible influence is of particular importance, introducing a possible bias in chronopharmacological studies. Thus, the aim of this study was to evaluate the effects of repeated caffeine administration on the circadian rhythms of heart rate "H," body temperature "T," and motor activity "A" in unrestrained rats maintained under controlled conditions (LD 12:12, light from 0600 to 1800 h) by using radiotelemetry transmitters. The study was divided into three 7-day observation spans: a 1-week control span "P1," a 1-week treatment span "P2," and a 1-week recovery span "P3." P1 was performed for assessing baseline measurements of H. T, and A. During P2, four rats received caffeine (25 mg/kg) at 0900 h, while four rats received saline in the same conditions every day of the observation span. H, T, and A were continuously monitored and plotted every 10 min. For P1, P2, and P3. a power spectrum analysis (Fourier transform) was applied to determine the dominant period of rhythmicity. If H, T, and A circadian rhythms were detected, the characteristics of these rhythms, i.e., mesors, amplitudes and acrophases, were determined by cosinor analysis, expressed as means +/- SEM and compared by analysis of variance. Our results indicated that caffeine did not suppress the circadian rhythmicity of H, T, and A, but significantly increased mesors and decreased amplitudes of the three rhythms and advanced acrophases of temperature and activity compared to the control group. PMID- 10463633 TI - The effect of labyrinthectomy on postural control of upside-down swimming catfish, Synodontis nigriventris, under pseudomicrogravity. AB - The catfish (Synodontis nigriventris) has a unique habitat of keeping an upside down posture under normal gravity. We examined its postural control under pseudomicrogravity generated artificially, and the effect of unilateral labyrinthectomy on the postural control. The stable swimming posture under pseudomicrogravity was observed in the upside-down swimming catfish but not in the catfish (Corydoras paleatus), which has normal swimming habitat. Furthermore, although S. nigriventris but not C. paleatus could keep the stable swimming posture under normal gravity condition after unilateral labyrinthectomy, the labyrinthectomized fishes could not keep it under pseudomicrogravity. Seven days after the operation, S. nigriventris alone partially recovered the ability to keep an upside-down swimming posture, and did completely, to the control level, 25 days after the operation. Furthermore, when S. nigriventris was under pseudomicrogravity in dark conditions, it showed disturbed swimming postures. These results suggest that the upside-down swimming catfish has superior ability of postural control depending on the labyrinth. PMID- 10463634 TI - Operant responding for dietary protein in the golden hamster (Mesocricetus auratus). AB - Golden hamsters housed in operant chambers over a period of weeks had ad lib access to a maintenance diet that was either nutritionally complete (NCMD) or protein-free (PFMD), and they were required to press a lever on a fixed-ratio (FR) schedule to obtain 20-mg high-protein pellets. As the FR requirement increased, hamsters maintained on the NCMD made fewer lever presses and ate fewer pellets, and at the highest FR levels, they earned very few pellets. In contrast, hamsters maintained on PFMD increased the number of lever presses as the FR requirement increased, and they only slightly reduced the number of pellets eaten. Even at the highest FR requirement levels, PFMD hamsters still derived an average of 11-12% of total calories from protein, a level of intake that is either adequate for adult hamsters, or very nearly so. Previous research has shown that hamsters make adaptive behavioural adjustments in response to time restricted access to dietary protein, and the present findings demonstrate that protein-restricted hamsters that must press a lever to obtain protein-rich pellets also make adaptive behavioural adjustments when challenged with increases in the FR requirement. PMID- 10463635 TI - Amygdala lesions produce analgesia in a novel, ethologically relevant acute pain test. AB - Acute pain tests using mechanical stimuli typically do not involve objects important in the evolutionary history of the subjects, and may fail to evaluate the contribution of biobehavioral defensive reactions to the total pain response. Spines are common structural defenses that protect plants and animals against predation. The present studies examined the reaction to contact with such natural, mechanical pain stimuli in the laboratory rat, utilizing a floor board with protruding pins located in the middle of a novel alley (the "fakir" test). Behavioral responses were characterized in 10-min tests (Experiment 1). Subjects showed voluntary contact with the pins followed by patterns of avoidance and risk assessment (stretch attend and stretch approach). Few subjects crossed the array of pins. The amygdala has been implicated in the perception of pain, particularly in stressful or fearful contexts. In Experiment 2, the fakir test was used to examine, concurrently, the effects of amygdala lesions on analgesiometric (frequency and duration of pin crossings) and anxiometric (risk assessment) measures. Large, bilateral, lesions of the amygdala significantly increased both the number of pin crossings and time spent on the pins without affecting the risk assessment measures. These findings suggest a possible dissociation between anxiety and pain perception with an important (nonaffective) role for the amygdala in the latter. PMID- 10463636 TI - Behavioral strategies in the cold: effects of feeding and nesting costs. AB - Animals can meet energetic challenges by acquiring or conserving energy. In the present experiment, we pitted these strategies against each other by housing rats in a cold environment and requiring them to bar press for food pellets and for access to a heated nest. Our question was how rats would exploit these resources to meet the added energetic demand of the cold when food was abundant or scarce. Results showed that rats' allocation of time and effort between competing activities was a function of demand. Feeding and nesting were partially substitutable in the cold: rats traded food for warmth and vice versa, economizing on both time and energy by adjusting their feeding and nesting patterns. Depending upon food abundance and the cost of nesting, rats acquired calories from food, conserved calories by nesting, and borrowed calories from the defense of body weight. PMID- 10463637 TI - Timidity in Japanese quail: effects of vitamin C and divergent selection for adrenocortical response. AB - Male Japanese quail chicks of two genetic lines selected for low (LS) or high (HS) adrenocortical responses to mechanical restraint were housed in mixed-line groups of 24 in four compartments of a multitier brooder battery at 20 days of age. Quail in two of the four compartments were given vitamin C (ascorbyl-2 polyphosphate, APP, 1 g L-ascorbic acid/L) solution for 48 h, whereas the other birds received untreated tap water as usual before they were tested at 23 days of age. At test, each quail was placed individually in a dark, sheltered compartment of an emergence box and allowed 1 min to acclimatise before a door was raised allowing access to an illuminated and exposed area. Vocalisation and the latencies to head and full emergence were then recorded to measure its fear levels. More LS quail vocalised than did HS ones. They also emerged more rapidly from the sheltered compartment into the illuminated one than HS birds. These findings further support our hypothesis that decreased fearfulness has accompanied genetic selection for reduced adrenocortical responsiveness. Treatment with APP reduced the latency to emerge fully into the exposed compartment, and there were no line x treatment interactions. These results suggest that vitamin C supplementation alleviated fearfulness, regardless of existing line differences in this behavioural trait. PMID- 10463638 TI - A comparison of period amplitude analysis and FFT power spectral analysis of all night human sleep EEG. AB - Zero-cross and zero-derivative period amplitude analysis (PAA) data were compared with power spectral analysis (PSA) data obtained with the fast Fourier transform in all-night sleep EEG from 10 subjects. Although PAA zero-cross-integrated amplitude showed good agreement with PSA power in 0.3-2 Hz, zero-cross analysis appears relatively ineffective in measuring 2-4 Hz and above waves. However, PAA zero-derivative measures of peak-trough amplitude correlated well with PSA power in 2-4 Hz. Thus, while PAA appears able to measure the entire EEG spectrum, the analytic technique should be changed from zero cross to zero derivative at about 2 Hz in human sleep EEG. PAA and PSA both demonstrate robust and interrelated across-night oscillations in three frequency bands: delta (0.3-4 Hz); sigma (12 16 Hz); and fast beta (20-10 Hz). The frequencies between delta and sigma, and between sigma and fast beta, did not show clear across-night oscillations using either method, and the two methods showed lower epoch-to-epoch agreement in these intermediate bands. The causes of this reduced agreement are not immediately clear, nor is it obvious which method gives more valid results. We believe that the three strongly oscillating frequency bands represent fundamental properties of the human sleep EEG that provide important clues to underlying physiological mechanisms. These mechanisms are more likely to be understood if their dynamic properties are preserved and measured naturalistically rather than being forced into arbitrary sleep stages or procrustean models. Both PAA and PSA can be employed for such naturalistic studies. PSA has the advantages of applying the same analytic method across the EEG spectrum and rests on more fully developed theory. Combined zero-cross and zero-derivative PAA demonstrates EEG oscillations that closely parallel those observed with spectral power, and the PAA measures do not rely on assumptions about the spectral composition of the signal. In addition, both PAA techniques can measure the relative contributions of wave amplitude and incidence to total power: These waveform characteristics represent different biological processes and respond differentially to a wide range of experimental conditions. PMID- 10463639 TI - Effects of social conflict on immune responses and E. coli growth within closed chambers in mice. AB - Social conflict has been shown to affect the neuroendocrine stress response in rodents. The current study was designed to characterize the effects of social conflict on leukocyte subset distribution and function as well as in vivo bacterial growth. Male DBA/2 mice implanted or not implanted with a closed chamber containing Escherichia coli were repeatedly challenged by temporary placement in the territory of a dominant CF-1 mouse five times a day for 2 consecutive days. Nonstressed animals were similarly handled, but were not exposed to social conflict. Effects on immune responses and E. coli growth were analyzed 13 h after the last social conflict session. Social conflict alone was associated with an increase in plasma corticosterone concentration and decreases in thymocyte numbers and splenocyte ability to proliferate in vitro in the presence of lipopolysaccharide (p < 0.05). After social conflict, immature CD4+CD8+ thymocytes decreased, whereas mature T cells increased (p < 0.05). In the presence of E. coli, social conflict induced a significant increase in plasma concentration of interleukin-1beta, and a decrease in the number of thymocytes and the percentage of CD4+CD8+ T cells in the thymus (p < 0.05). In addition to the lymphocyte subpopulation changes observed with social conflict alone, the proportion of CD3+ and major histocompatibility complex (MHC) class II IAd+ cells were significantly higher in stressed mice implanted with a closed chamber containing E. coli (p < 0.05). Social conflict tended to favor E. coli growth in the closed chamber, indicating possible direct bacterial-neuroendocrine hormone interactions. Taken together, these results suggest that stress may modulate the host immune response by altering both bacterial growth and resistance to infection. PMID- 10463640 TI - Cyclic estradiol treatment normalizes body weight and test meal size in ovariectomized rats. AB - We tested whether cyclic estradiol treatment, like continuous estradiol treatment, is sufficient to normalize meal size and body weight in ovariectomized rats. In Experiment 1, adult Long-Evans rats were ovariectomized and subcutaneously injected with 0, 0.2, or 2.0 microg estradiol benzoate (EB) in sesame oil each Tuesday and Wednesday. Oil-treated ovariectomized rats gained more weight during 4 weeks of ad lib feeding (48 +/- 5 g) than intact rats (16 +/ 1 g, p < 0.01). Cyclic treatment with 2.0 microg EB normalized weight gain (11 +/- 2 g). During the next week, plasma samples were assayed for estradiol. Cyclic treatment with 2.0 microg EB produced excursions of plasma estradiol that appeared similar to those of intact, cycling rats: estradiol level reached 190 +/ 60 pmol/L after the second EB injection before decreasing to undetectable levels (<30 pmol/L) by cycle end. In Experiment 2, test meal sizes after overnight food deprivation were measured. Cyclic treatment with 2.0 microg EB produced both tonic (i.e., at cycle onset, meal size was smaller in estradiol-treated than oil treated rats) and phasic (i.e., meal size was smaller late in the EB-treatment cycle than early in it) decreases in meal size. Thus, a weekly cyclic regimen of estradiol treatment that produces changes in plasma estradiol concentration similar to those in intact cycling rats is sufficient to produce the body weight and meal size patterns that characterize normal hypothalamic-pituitary-gonadal function. PMID- 10463641 TI - Prostaglandin E2-induced fever in young and old Long-Evans rats. AB - Aging is associated with a blunted or absent fever response to naturally occurring infections or to the peripheral administration of bacterial products and proinflammatory cytokines. We have recently shown that old Long-Evans rats are not defective in their capacity to develop a fever in response to brain administration of interleukin-1beta (IL-1beta). Here, we investigated the fever response of young (3-5-month) and old (24-26-month) Long-Evans rats to the intracerebroventricular (i.c.v.) microinfusion of prostaglandin E2 (PGE2), a final common mediator for the production of fever in the brain. Core body temperature was monitored by telemetry in freely moving rats. i.c.v. administered PGE2 (100 ng) induced comparable increases in body temperature in young and old Long-Evans rats. In the two groups, PGE2-induced fever was similar both in latency-to-peak fever and maximal fever response. These data, and the previous data on IL-1beta, demonstrate that the brains of old and young rats are similar with respect to fever generation in response to the i.c.v. administration of two classes of immunomodulators. PMID- 10463642 TI - The effects of electroconvulsive shock on retinal activity. AB - Two experiments are reported examining the effects of electroconvulsive shock (ECS) on the electroretinogram (ERG) and the retinal oscillatory potentials (OPs) in the albino rat. Immediately after the induction of generalised seizure activity, both the ERG and the OPs were always preserved basically intact, despite minor alterations to their waveforms. In Experiment 1, it was found that small changes in amplitude of the ERG were recorded following ECS, but these were most likely artifactual. The only other notable finding was a temporary decrease in latency of the b-wave of the ERG. In Experiment 2, a slight overall attenuation in the amplitude of the OPs was observed. This was associated with a paradoxical decrease in the latencies of all three OP subcomponents similar to that found for the ERG b-wave. It is concluded that ECS does not interfere to any marked extent with either the transduction of the visual signal or its processing within the various retinal layers. This implies that the blockade of the afferent volley that occurs following ECS must be confined to the optic pathway or to the occipital cortex itself. PMID- 10463643 TI - Medial tibial pain: a dynamic contrast-enhanced MRI study. AB - The purpose of this study was to compare the sensitivity of different magnetic resonance imaging (MRI) sequences to depict periosteal edema in patients with medial tibial pain. Additionally, we evaluated the ability of dynamic contrast enhanced imaging (DCES) to depict possible temporal alterations in muscular perfusion within compartments of the leg. Fifteen patients with medial tibial pain were examined with MRI. T1-, T2-weighted, proton density axial images and dynamic and static phase post-contrast images were compared in ability to depict periosteal edema. STIR was used in seven cases to depict bone marrow edema. Images were analyzed to detect signs of compartment edema. Region-of-interest measurements in compartments were performed during DCES and compared with controls. In detecting periosteal edema, post-contrast T1-weighted images were better than spin echo T2-weighted and proton density images or STIR images, but STIR depicted the bone marrow edema best. DCES best demonstrated the gradually enhancing periostitis. Four subjects with severe periosteal edema had visually detectable pathologic enhancement during DCES in the deep posterior compartment of the leg. Percentage enhancement in the deep posterior compartment of the leg was greater in patients than in controls. The fast enhancement phase in the deep posterior compartment began slightly slower in patients than in controls, but it continued longer. We believe that periosteal edema in bone stress reaction can cause impairment of venous flow in the deep posterior compartment. MRI can depict both these conditions. In patients with medial tibial pain, MR imaging protocol should include axial STIR images (to depict bone pathology) with T1-weighted axial pre and post-contrast images, and dynamic contrast enhanced imaging to show periosteal edema and abnormal contrast enhancement within a compartment. PMID- 10463644 TI - Evaluation of cystic ovarian lesions using apparent diffusion coefficient calculated from reordered turboflash MR images. AB - Reordered snapshot fast low-angle shot images with, and without, diffusion perfusion gradients were used for the evaluation of contents of cystic ovarian lesions. Sonographically detected 51 cystic ovarian lesions (13 endometrial cysts, 17 ovarian cysts, 7 serous cystadenomas, 6 mucinous cystadenomas, 8 malignant cystic ovarian tumors) were studied. T1- and T2-weighted images, reordered snapshot fast low-angle shot images with and without diffusion perfusion gradients (b = 106 and 0 s/mm2, respectively) were obtained. Using these images, apparent diffusion coefficients (ADCs) were calculated in the cystic contents of these lesions. Endometrial cysts and malignant cystic ovarian tumors showed lower ADC values than ovarian cysts, serous cystadenomas and mucinous cystadenomas (p < 0.02). There was no distinct ADC difference among ovarian cysts, serous cystadenomas, mucinous cystadenomas (p > 0.2). In conclusion, diffusion-weighted magnetic resonance imaging is possible to be useful to evaluate cystic contents of ovarian lesions. PMID- 10463645 TI - MRI appearance of placenta percreta and placenta accreta. AB - The purpose of this paper is to describe the magnetic resonance imaging (MR) features of placenta accreta and percreta. We retrospectively reviewed MRI findings in four cases of placenta accreta/percreta to determine features which assist in identifying the presence and extent of placental implantation abnormality. All patients had ultrasound (US) examinations. Pathologic correlation was available in all cases. There were two cases of placenta percreta and two cases of placenta accreta. All cases were treated by hysterectomy. In the two cases of placenta percreta, the placenta demonstrated transmural extension through the uterus (percreta) on MRI. In the two cases of placenta accreta, the location of thinning in the uterine wall correlated with the location of placental invagination into the myometrium at pathology. US correlation was available in all four cases. Gray scale US did not demonstrate placental invasion in any of the four cases of placenta accreta/percreta, however, in two of three cases in which color Doppler was performed, there was flow at the uterine margin suspicious for implantation abnormality. In conclusion, MRI is useful for identifying the presence and extent of placenta accreta/percreta. PMID- 10463646 TI - Magnetic resonance imaging of postpartum pelvic hematomas: early experience in diagnosis and treatment planning. AB - The purpose of this study was to determine the utility of magnetic resonance imaging (MRI) in detecting and localizing clinically non-apparent postpartum pelvic hematomas, and to describe the therapeutic implications of MRI in these patients. MRI examinations were performed on seven women with pelvic pain and/or fullness following difficult vaginal deliveries. None had clinically evident hematomas, and none were scheduled for surgery. Hematomas in the pelvis were identified with conventional T1-weighted, and conventional and fast spin echo T2 weighted sequences. Intravenous contrast was not used. Pelvic hematomas were identified in contiguity with the vagina, cervix, and bladder, within the broad ligament, and in the presacral space. In two patients, hematomas were confined to the perivaginal and pericervical regions. In three patients, perivaginal hematomas extended between the double layers of the broad ligaments. In one patient, a perivaginal hematoma extended into the perivesical space. In one patient, a hematoma was identified only within the presacral space. Based on MRI as the only contributory imaging study, five patients were treated successfully with invasive means and two patients were managed successfully with conservative means. MRI successfully detects and localizes postpartum hematomas, information that often is unavailable from the clinical examination. This information facilitates decisions regarding the need for intervention and the appropriate type of procedure when intervention is necessary for this potentially life threatening problem. We advocate the use of MRI to evaluate patients at risk for postpartum hemorrhage following difficult vaginal deliveries. PMID- 10463647 TI - Short-term evolution of individual enhancing MS lesions studied with magnetization transfer imaging. AB - We performed serial monthly magnetization transfer (MT) imaging to evaluate the prevalence and evolution of structural changes in individual enhancing lesions from patients with multiple sclerosis (MS). Every 4 weeks for 3 months, we obtained dual echo, magnetization transfer (MT) imaging and, 5 min after SD (0.1 mmol/kg) gadolinium-DTPA injection, T1-weighted scans from 10 patients with early relapsing-remitting MS. We measured the MT ratio (MTR) of enhancing lesions seen on the entry scans on co-registered quantitative MTR images at entry and during the follow up. Fourty-two enhancing lesions were identified on the entry scans. According to the "maximal random fluctuation" detected for the normal-appearing white matter MTR values, 16 (38%) lesions were classified as "increasing MTR" lesions, 21 (50%) as "stable MTR" lesions, and 5 (12%) as "decreasing MTR" lesions. The classification of the lesions after the first month of follow up strongly predicted the classification at the end of the follow up (chi squared = 20.35, p = 0.0004). These results indicate that the enhancing lesion population in MS is heterogeneous, and that reparative mechanisms occurring after blood brain barrier opening are not efficient in only a minority of the enhancing lesions from patients with early relapsing-remitting MS. PMID- 10463648 TI - Correlation between enhancing lesion number and volume on standard and triple dose gadolinium-enhanced brain MRI scans from patients with multiple sclerosis. AB - We investigated the correlations between numbers and volumes of multiple sclerosis (MS) lesions enhancing on standard dose (SD) and triple dose (TD) gadolinium (Gd)-enhanced brain magnetic resonance imaging (MRI) scans, to clarify whether the measurement of enhancing lesion volumes or the use of TD MRI give additional information which can not be obtained by counting enhancing lesions on SD scans. SD and TD Gd-enhanced brain MRI scans were obtained every month for three months from 40 MS patients. The numbers of total and new enhancing lesions were counted, and the total volumes of enhancing lesions were measured from each of the four scans obtained with the two techniques. Univariate correlations between enhancing lesion numbers and volumes were assessed. The numbers of total and new enhancing lesions seen either on SD or TD scans were significantly correlated (r = 0.91 and 0.93, respectively). The numbers and volumes of total enhancing lesions were significantly correlated on both SD (r = 0.90), and TD (r = 0.89) scans. Moderate correlations were found between the total number of enhancing lesions on SD scans and the average difference between TD and SD scans for total enhancing lesion number (r = 0.66), and between the number of new enhancing lesions on SD scans and the average difference between TD and SD scans for new enhancing lesion number (r = 0.50). Our findings indicate that, both on SD and TD MRI, the counts and the volumes of total and new enhancing lesions are highly correlated, and that lesion counting may suffice to monitor MS activity. On the contrary, this study confirms the usefulness of TD MRI for a more complete assessment of the acute changes occurring in MS patients. PMID- 10463649 TI - R'2 measured in trabecular bone in vitro: relationship to trabecular separation. AB - Measurement of key parameters of the microstructure of trabecular bone is critical to the study of osteoporosis and bone strength. Density based methods cannot provide this information, and give only the total amount of bone present, and not its arrangement. Magnetic resonance imaging has shown the potential to provide information related to the microarchitecture of the trabecular bone matrix. Twelve samples (8 x 8 x 8 mm3 bone cubes) were cut from sheep vertebrae such that the trabeculae ran either parallel or perpendicular to each face. Detailed measurements of the structure of these bone cubes were made by histomorphometry, and compared to R'2 and R*2 measured with a spin and gradient echo sequence, Partially Refocused Interleaved Multiple Echo, at 1.5 Tesla. The precision of the R'2 measurement (% coefficient of variation) was 8.7+/-5.1, and 7.7+/-4.3 for R*2. Uncorrected values of R'2 and R*2 were significantly correlated to density measured by quantitative computed tomography (r = 0.87, p = 0.0005, and r = 0.90, p = 0.0002, respectively), and trabecular bone area measured by histomorphometry (r = 0.80, p = 0.002, and r = 0.83, p = 0.0008, respectively). Density correction was effected by imaging the same slice of bone in two orientations (90 degrees and 0 degrees ) to the main magnetic field. For both R'2 and R*2 there was a significant difference between measurements in the 90 degrees and 0 degrees orientations (p < 0.01). The difference between the two values was used, and termed R'2net or R*2net. The net parameters were independent of bone mass. R'2net and R*2net were significantly correlated to trabecular separation (p < 0.05) with r = -0.58 and r = -0.62, respectively. These results demonstrate the ability of magnetic resonance imaging to characterize a key measure of the trabecular microstucture. An increase in trabecular separation has important biomechanical consequences in osteoporosis. This result also strengthens the hypothesis that the sensitivity of R'2 to osteoporosis-related bone changes is due to magnetic susceptibility effects in which rapid transitions between bone and marrow create local magnetic field inhomogeneities that result in an increase in R'2 values. PMID- 10463650 TI - Magnetic resonance T2* measurements of the normal human lung in vivo with ultra short echo times. AB - The objective of this study was to measure T2* values of the normal human lung in vivo during breathhold using a rapid gradient-echo sequence with ultra-short echo times (TE). A sagittal slice of the right lung was imaged in six volunteers with various TE ranging from 0.5 ms to 5 ms using a clinical 1.5 Tesla MR scanner. T2* values were calculated in a region of interest in the dependent and non-dependent lung. In the dependent lung, T2* values of 1.1 ms+/-0.15 ms were measured, and in the non-dependent lung, 0.86 ms+/-0.11 (p < 0.01). T2* measurements of the normal human lung during breathhold are feasible with a clinical MR unit. The short T2* values require the use of very short TE times (< 2.5 ms) in gradient-echo sequences to obtain adequate signal intensity from lung tissue. PMID- 10463651 TI - 3D spin-lock imaging of human gliomas. AB - We investigated whether the simultaneous use of paramagnetic contrast medium and 3D on-resonance spin lock (SL) imaging could improve the contrast of enhancing brain tumors at 0.1 T. A phantom containing serial concentrations of gadopentetate dimeglumine (Gd-DTPA) in cross-linked bovine serum albumin (BSA) was imaged. Eleven patients with histologically verified glioma were also studied. T1-weighted 3D gradient echo images with and without SL pulse were acquired before and after a Gd-DTPA injection. SL effect, contrast, and contrast to-noise ratio (CNR) were calculated for each patient. In the glioma patients, the SL effect was significantly smaller in the tumor than in the white and gray matter both before (p = 0.001, p = 0.025, respectively), and after contrast medium injection (p < 0.001, p < 0.001, respectively). On post-contrast images, SL imaging significantly improved tumor contrast (p = 0.001) whereas tumor CNR decreased slightly (p = 0.024). The combined use of SL imaging and paramagnetic Gd-DTPA contrast agent offers a modality for improving tumor contrast in magnetic resonance imaging (MRI) of enhancing brain tumors. 3D gradient echo SL imaging has also shown potential to increase tissue characterization properties of MR imaging of human gliomas. PMID- 10463652 TI - Correction of eddy current-induced artefacts in diffusion tensor imaging using iterative cross-correlation. AB - Geometric distortions of echo-planar images produced by the strong eddy currents present in the diffusion tensor imaging experiment are a major confound to the accurate quantification of diffusion coefficients, and measures of diffusion anisotropy based upon them. Here we investigate how the method of iterative cross correlation (ICC) of baseline and diffusion-weighted images (DWIs) originally proposed by Haselgrove and Moore (Magn. Reson. Med. 36:960-964; 1996) can be extended to correct high b-value DWIs, without the need for extrapolation of distortion parameters determined from low b-value images. Monte Carlo simulations of synthetic brain images show that the maximum value of the trace of the b matrix, Tr(b), at which distorted DWIs can be accurately corrected by direct comparison with the undistorted baseline image is approximately 300 s mm(-2). Removal of the cerebrospinal fluid signal greatly extends this value of Tr(b) (up to approximately 2000 s mm(-2)), thereby allowing direct comparison of baseline and distorted images. The use of ICC distortion parameters determined from separate calibrations of water phantom images is also investigated, and found to be effective in correcting geometric distortions observed in the DWIs collected as part of a human volunteer diffusion tensor imaging study. This work suggests that distorted DWIs acquired at high values of b may be corrected using the ICC algorithm without collecting additional low b-value images, thus allowing simplified methods of measuring the apparent diffusion tensor D, based on collecting a small number of DWIs, to be implemented in quantitative patient examinations. PMID- 10463653 TI - Multicentre magnetic resonance texture analysis trial using reticulated foam test objects. AB - Texture analysis in magnetic resonance imaging has the ability to provide useful diagnostic information with respect to the discrimination of disease states of a single tissue or the separation of different tissues. However, for widespread use it is necessary to determine how texture measurements carried out in one center relate to those carried out in another. To this end, a multicentre trial has been performed where reticulated foam test objects have been scanned in six European centers according to a fixed protocol. It has been concluded that texture measurements are not transportable between centers. Principal component models calculated from the texture parameters collected in one center do not fit the data collected in another. Further trials are to investigate whether the reticulated foam test objects may be used to normalize tissue texture data collected in different centers. PMID- 10463654 TI - Interobserver reproducibility of quantitative cartilage measurements: comparison of B-spline snakes and manual segmentation. AB - The objective of this work was to develop a segmentation technique for thickness measurements of the articular cartilage in MR images and to assess the interobserver reproducibility of the method in comparison with manual segmentation. The algorithm is based on a B-spline snakes approach and is able to delineate the cartilage boundaries in real time and with minimal user interaction. The interobserver reproducibility of the method, ranging from 3.3 to 13.6% for various section orientations and joint surfaces, proved to be significantly superior to manual segmentation. PMID- 10463655 TI - T1rho dispersion of rat tissues in vitro. AB - The purpose of this study was to demonstrate T1rho dispersion in different rat tissues (liver, brain, spleen, kidney, heart, and skeletal muscle), and to compare the 1/T1rho data to previous 1/T1 data and magnetization transfer of rat tissues at low (0.1 T) B0 field. The 1/T1rho dispersion showed a fairly similar pattern in all tissues. The highest 1/T1rho relaxation rates were seen in liver and muscle followed by heart, whereas the values for spleen, kidney, and brain were quite similar. Compared to 1/T2 relaxation rate, the greatest difference was seen in liver and muscle. The rank order 1/T1rho value at each locking field B1 was the same as the transfer rate of magnetization from the water to the macromolecular pool (Rwm) for liver, muscle, heart, and brain. The potential value T1rho imaging is to combine high T1 contrast of low field imaging with the high signal to noise ratio of high static field imaging. When the T1rho value for a given tissue is known, the contrast between different tissues can be optimized by adjusting the locking time TL. Further studies are encouraged to fully exploit this. Targets for more detailed research include brain infarct, brain and liver tumors. PMID- 10463656 TI - Effects of hyperthermia on bioenergetic status and phosphorus T1S in human melanoma xenografts monitored by 31P-MRS. AB - Conventional hyperthermia enhances tumor response to radiotherapy through thermal cell inactivation and vascular shut-down, whereas mild hyperthermia potentiates the effect of radiotherapy by improving tumor oxygenation. The work reported here was aimed at investigating whether 31P-magnetic resonance spectroscopy (31P-MRS) measurements of tumor bioenergetic status; i.e., the (PCr + NTPbeta)/Pi resonance ratio, and/or the spin lattice relaxation times, T1s, of the Pi and NTPbeta resonances can be used to distinguish between the effects of conventional and mild hyperthermia. BEX-t human melanoma xenografts were treated at 43.0 degrees C for 15 or 60 min, and bioenergetic status and T1s were measured as function-of time after treatment. Hyperthermia-induced effects on tumor blood flow was measured by using the 86Rb uptake method. The morphology of the capillary network in treated and untreated tumors was studied by histologic examination. Tumors treated for 15 min showed increased blood flow and dilated capillaries, whereas tumors treated for 60 min showed decreased blood flow and capillary occlusions; i.e., 43.0 degrees C for 15 min was a treatment consistent with mild hyperthermia and 43.0 degrees C for 60 min was consistent with conventional hyperthermia treatment of BEX-t tumors. Bioenergetic status increased after treatment at 43.0 degrees C for 15 min, and decreased after treatment at 43.0 degrees C for 60 min, similar to the blood flow. Likewise, the T1 of the Pi resonance increased after treatment at 43.0 degrees C for 15 min, and decreased after treatment at 43.0 degrees C for 60 min. The T1 of the NTPbeta resonance showed a similar change as the T1 of the Pi resonance, but less pronounced. Consequently, 31P-MRS measurements of tumor bioenergetic status and the T1 of the Pi resonance may perhaps be utilized to distinguish between vascular effects of mild and conventional hyperthermia. PMID- 10463657 TI - A post-processing technique for displaying vessels from routine fast-spin-echo images: MRI-derived angiography. AB - Fast-spin-echo magnetic resonance (MR) images are routine components of a standard MR brain examination. On these images, blood vessels are visible as black flow void. We report that by applying an enhancement filter to a stack of routine fast-spin-echo MR images, projected angiographic images can be generated. The vascular detail in the projected image is similar to that observed in a phase contrast image. In addition to its advantage in obtaining vessel information from routine images, the proposed post-processing technique is fast, easy to implement and completely automatic. These images provide additional vessel information that is useful when MR angiography is unavailable or as an aid in planning dedicated MR angiographic studies. PMID- 10463658 TI - A modified fuzzy clustering algorithm for operator independent brain tissue classification of dual echo MR images. AB - Methods for brain tissue classification or segmentation of structural magnetic resonance imaging (MRI) data should ideally be independent of human operators for reasons of reliability and tractability. An algorithm is described for fully automated segmentation of dual echo, fast spin-echo MRI data. The method is used to assign fuzzy-membership values for each of four tissue classes (gray matter, white matter, cerebrospinal fluid and dura) to each voxel based on partition of a two dimensional feature space. Fuzzy clustering is modified for this application in two ways. First, a two component normal mixture model is initially fitted to the thresholded feature space to identify exemplary gray and white matter voxels. These exemplary data protect subsequently estimated cluster means against the tendency of unmodified fuzzy clustering to equalize the number of voxels in each class. Second, fuzzy clustering is implemented in a moving window scheme that accommodates reduced image contrast at the axial extremes of the transmitting/receiving coil. MRI data acquired from 5 normal volunteers were used to identify stable values for three arbitrary parameters of the algorithm: feature space threshold, relative weight of exemplary gray and white matter voxels, and moving window size. The modified algorithm incorporating these parameter values was then used to classify data from simulated images of the brain, validating the use of fuzzy-membership values as estimates of partial volume. Gray:white matter ratios were estimated from 20 twenty normal volunteers (mean age 32.8 years). Processing time for each three-dimensional image was approximately 30 min on a 170 MHz workstation. Mean cerebral gray and white matter volumes estimated from these automatically segmented images were very similar to comparable results previously obtained by operator dependent methods, but without their inherent unreliability. PMID- 10463659 TI - Moisture gradient vector calculation as a new method for evaluating NMR images of corn (Zea Mays L.) kernels during drying. AB - Magnetic resonance imaging (MRI) was used to study the moisture migration inside single corn kernel during artificial drying. MRI data were taken every half an hour during drying while the intact kernel was kept inside the magnet. The moisture distribution was not uniform inside the kernel during drying, with the fastest moisture loss being detected in the endosperm, whilst the scutellum maintained the water. Resultant gradient vectors were visualized, as a new evaluation method, from each pixel of the subtraction of segmented MRI images showing the movement of proton density inside the kernel. The data matrix containing the gradient vectors can be stored for latter use as input parameters and to check the mass transfer models. PMID- 10463660 TI - Probe efficiency improvement with remote and transmission line tuning and matching. AB - In certain nuclear magnetic resonance (NMR) applications in which accessibility to the gantry is limited, performing optimal tuning and matching represents a major problem. Here, we discuss a method of tuning NMR probe circuits and matching their impedances which uses cables with different impedance values. This simple but efficient method may be advantageous compared with much more difficult perfect tuning and matching. PMID- 10463661 TI - MR relaxometry on a whole-body imager: quality control. AB - Long-term monitoring of the average proton relaxation time T2 of phantoms measured on a Siemens MR whole-body imager showed very good repeatability and reproducibility. The repeatability (short-term precision) and reproducibility (long-term precision) of the average values of a relaxation time T2 approximately/= 81 ms, obtained by a standard 16-echo CPMG pulse sequence, were 2.6% and 9.7%, respectively. The Siemens Vision imager proved to be a suitable tool for T2 evaluation in vivo. Quality control was performed using the techniques of control diagrams developed by Shewhart, which proved to be an appropriate method for continuous quality control of relaxation time determination. PMID- 10463662 TI - Islet cell tumor of the pancreas associated with tumor thrombus in the portal vein. AB - We report the MR findings of a 70-year-old man with an islet cell tumor that diffusely involved the body of the pancreas associated with enhancing portal vein tumor thrombus and cavernous transformation. The diffusely infiltrative tumor mass was best shown on early post gadolinium spoiled gradient echo. The tumor thrombus enhanced intensely on early post gadolinium images and was also well shown on true FISP (Fast Imaging with Steady State Precession) images. The extent of liver metastases was best shown on fat suppressed T2-weighted images. The most unusual finding was tumor thrombus involving the SMV and portal vein. PMID- 10463663 TI - Malaria diagnosis. PMID- 10463664 TI - Acceleration of viral replication and up-regulation of cytokine levels by antimalarials: implications in malaria-endemic areas. AB - Antimalarial drugs are widely used in malaria endemic areas, both for chemoprophylaxis and also empirically to treat patients presenting with fever. Previously, we have reported that chloroquine enhances the severity of Semliki forest virus (SFV) and encephalomyocarditis virus infection. The studies presented herein show that a broad spectrum of antimalarial drugs augmented the replication of SFV in mice, concomitant with greater tissue damage and up regulation of mRNA levels of various inflammatory cytokine genes, including interleukin-1 receptor antagonist (IL-1Ra), II-1alpha, IL-1beta, IL-6, IL-12p40, and interferon-gamma inducing factor. Furthermore, chloroquine enhances IL-1Ra production in RAW cells in vitro. Since IL-1Ra is known to be up-regulated in a number of viral infections, we propose that a further enhancement of its expression by antimalarials may be responsible for the increased severity of viral infection in our studies. Thus, the widespread use of antimalarials in malaria-endemic areas may predispose the population to viral infections. Further studies are in progress to delineate mechanism(s) involved in cytokine up regulation and acceleration of viral replication. PMID- 10463665 TI - In vitro activity of dihydroartemisinin against clinical isolates of Plasmodium falciparum in Yaounde, Cameroon. AB - The in vitro activities of dihydroartemisinin (the biologically active metabolite of artemisinin derivatives), chloroquine, monodesethylamodiaquine (the biologically active metabolite of amodiaquine), quinine, mefloquine, halofantrine, and pyrimethamine were assessed in 65 African isolates of Plasmodium falciparum from Yaounde, Cameroon using an isotopic microtest. The 50% inhibitory concentration (IC50) values for dihydroartemisinin were within a narrow range from 0.25 to 4.56 nM, with a geometric mean of 1.11 nM (95% confidence interval = 0.96-1.28 nM). Dihydroartemisinin was equally active (P > 0.05) against the chloroquine-sensitive isolates (geometric mean IC50 = 1.25 nM, 95% confidence interval = 0.99-1.57 nM) and the chloroquine-resistant isolates (geometric mean IC50 = 0.979 nM, 95% confidence interval = 0.816-1.18 nM). A significant positive correlation was observed between the responses to dihydroartemisinin and mefloquine (r = 0.662) or halofantrine (r = 0.284), suggesting in vitro cross-resistance. There was no correlation between the responses to dihydroartemisinin and other antimalarial drugs. PMID- 10463666 TI - In vitro and in vivo activity of amphotericin B-lipid formulations against experimental Trypanosoma cruzi infections. AB - The activities of four amphotericin B formulations, Fungizone, AmBisome, Amphocil, and Abelcet, were compared in vitro and in vivo against Trypanosoma cruzi infections. In vitro, Fungizone and Amphocil were highly active against T. cruzi Y strain amastigotes in macrophages with 50% effective dose (ED50) values of 0.027-0.028 microg/ml, which were 7-fold and 42-fold more active than AmBisome and Abelcet, respectively. In vitro activities of all formulations against T. cruzi amastigotes in Vero cells were similar, with ED50 values in the range of 2.0-4.2 microg/ml. Acute infections of the T. cruzi Y strain in BALB/c mice were suppressed in all animals by a single 25 mg/kg dose of the liposomal formulation AmBisome. At the same dose, the two other lipid formulations, Amphocil and Abelcet, increased the survival rate but did not suppress infection in all animals. PMID- 10463667 TI - Clinical analysis and parasite genetic diversity in human immunodeficiency virus/Chagas' disease coinfections in Brazil. AB - To evaluate the possible role of parasitemia on Chagas' disease reactivation in Chagas' disease/human immunodeficiency virus (HIV) coinfection cases and the impact of HIV coinfection on Trypanosoma cruzi genetic diversity, 71 patients with Chagas' disease (34 HIV+ and 37 HIV-) were surveyed. Moreover, 92 T. cruzi stocks from 47 chronic chagasic patients (29 HIV+ and 18 HIV-) were isolated and analyzed by multilocus enzyme electrophoresis and a random amplified polymorphic DNA procedure. High parasitemia appeared to play a major role in cases of Chagas' disease reactivation. In HIV+ patients, the genetic diversity and population structure (clonality) of T. cruzi was similar to that previously observed in HIV- patients, which indicates that immunodepression does not modify drastically genotype repartition of the parasite. There was no apparent association between given T. cruzi genotypes and specific clinical forms of Chagas' disease/HIV associations. PMID- 10463668 TI - Application of cardiac gated magnetic resonance imaging in murine Chagas' disease. AB - To evaluate the role of gated cardiac magnetic imaging resonance (MRI) in Chagas' disease, we infected mice with Trypanosoma cruzi (Brazil strain). Two models were chosen for study, the CD1 and the inducible nitric oxide synthase knockout (NOS2 /-) mice. Infection of CD1 mice was associated with a significant increase in the right ventricular inner diameter (RVID) that was reversed in some mice by verapamil. Expression of cardiac NOS2 has been associated with myocardial dysfunction. Therefore, we evaluated chagasic cardiomyopathy in NOS2-/- and syngeneic wild type (WT) mice. Infected WT mice exhibited an increase in RVID in the acute phase (< 60 days postinfection) that was more marked during chronic infection (>100 days postinfection). Chronically infected NOS2-/- mice had an increase in RVID. The RVID in infected WT mice was greater than in NOS2-/- mice. These data demonstrate that MRI is a useful tool in the serial evaluation of the heart in murine Chagas' disease. In addition, it supports the notion that the NOS2-/-/NO pathway may contribute to the pathogenesis of murine chagasic cardiomyopathy. PMID- 10463669 TI - Urine circulating soluble egg antigen in relation to egg counts, hematuria, and urinary tract pathology before and after treatment in children infected with Schistosoma haematobium in Kenya. AB - A cohort of 117 school children infected with Schistosoma haematobium was followed-up after therapy with praziquantel (0, 2, 4, 6, 12, and 18 months) and various infection and morbidity parameters (egg counts, hematuria, soluble egg antigen [SEA] in urine, and ultrasonography-detectable pathology) were quantified. At the onset of the study, 97% of the children were positive for S. haematobium with a geometric mean egg count of 45.7 eggs/10 ml of urine. Eighty one percent of the children were positive for SEA in urine with a geometric mean SEA concentration of 218.8 ng/ml of urine. Ninety-two percent and 56% of the children were microhematuria positive and macrohematuria positive, respectively. Two months after treatment, all infection and morbidity indicators had significantly decreased. Reinfection after treatment as determined by detection of eggs in urine was observed by four months post-treatment while the other parameters remained low. The clearance of SEA was slower than that of egg counts while pathology resolved at an even slower pace. Levels of SEA and egg output showed similar correlations with ultrasound detectable pathology; these correlations were better than the correlation between hematuria and pathology. PMID- 10463670 TI - Sequence analysis of the small RNA segment of guinea pig-passaged Pichinde virus variants. AB - The established animal model for Lassa fever is based on the new world arenavirus Pichinde (PIC). Natural isolates of PIC virus are attenuated in guinea pigs, but serial guinea pig passage renders them extremely virulent in that host. We have compared the nucleotide sequences of the small RNA segments of two attenuated, low-passage variants of the PIC virus Munchique strain (CoAn 4763) and two virulent, high-passage derivatives. Missense mutations in the glycoprotein precursor (GPC) gene at codons GPC-119, GPC-140, and GPC-164 and the nucleoprotein gene (NP) codons NP-35 and NP-374 were most closely associated with virulence. Codon GPC-140 is predicted to represent a region of peak hydrophilicity of the glycoprotein 1 (GP1); it is conceivable that mutations at this site could influence virulence by altering B cell epitopes or virus attachment protein conformation. PMID- 10463671 TI - Cytokine production in rhesus monkeys infected with Plasmodium coatneyi. AB - Plasmodium coatneyi infection in rhesus monkeys has been used as a model for studying human malaria. Cytokine production in this model, however, has so far not been examined. In this study, four rhesus monkeys were infected with P. coatneyi, with another four animals serving as uninfected controls. Blood samples were taken for the determination of daily parasitemia, and cytokine and prostaglandin E2 (PGE2) levels at days 0, 3, 5, 7, and 10. All inoculated animals became infected, with synchronized appearance of ring-stage parasites. Infected monkeys had increased plasma levels of proinflammatory cytokines (interleukin 1beta, interferon-gamma, and tumor necrosis factor-alpha) during the late stage of the infection. They also had increased production of ciliary neurotrophic factor. In conjunction with the production of proinflammatory cytokines, infected monkeys also had gradual increases in the production of PGE2. A continued definition of the P. coatneyi/rhesus monkey animal model should be useful for the elucidation of the immunopathogenesis of human malaria. PMID- 10463672 TI - Mitochondrial DNA variation among Anopheles albimanus populations. AB - Barriers to gene flow between Pacific and Atlantic coast populations of Anopheles albimanus were reported in an earlier study of variation in the intergenic spacer of the nuclear ribosomal DNA. We examined the distribution of mitochondrial DNA haplotypes among A. albimanus populations to test for gene flow barriers with an independent genetic marker. A region of the NADH dehydrogenase subunit 5 gene was amplified by the polymerase chain reaction (PCR) in 1,105 mosquitoes collected from 16 locations in Guatemala and in single collections from Mexico, Honduras, Nicaragua, Costa Rica, Panama, Colombia, and Venezuela. The PCR products were tested for variation using single strand conformation polymorphism analysis and 45 haplotypes were detected. Haplotype frequencies did not vary between coasts in Guatemala. Populations within approximately 200 km of one another were panmictic. However, at distances > 200 km, FST and geographic distances were correlated suggesting that populations are isolated by distance. PMID- 10463673 TI - Plasmodium falciparum mdr1 mutations and in vivo chloroquine resistance in Indonesia. AB - Mutations in the Pfmdr1 gene are reported to be associated with chloroquine resistance in some Plasmodium falciparum isolates. A polymerase chain reaction/restriction fragment length polymorphism method was used for the detection of Pfmdr1 mutations in chloroquine-resistant field isolates of P. falciparum collected in Irian Jaya. The frequency of Pfmdr1 mutations was significantly higher in chloroquine-resistant P. falciparum parasites than background frequencies observed in the same location. The 7G8 mutation was identified in some parasites although always in a mixed genotype status. Chloroquine-resistant P. falciparum specimens were characterized using the World Health Organization 28-day criteria, supplemented by demonstrating adequate chloroquine absorption and genetic analysis. PMID- 10463674 TI - Lack of an association between the ASN-108 mutation in the dihydrofolate reductase gene and in vivo resistance to sulfadoxine/pyrimethamine in Plasmodium falciparum. AB - Sulfadoxine/pyrimethamine (SP) is considered an alternative treatment for acute uncomplicated malaria caused by Plasmodium falciparum resistant to chloroquine. However, the appearance of resistance to this drug has been reported since its initial use in Colombia. Molecular analysis of the dihydrofolate reductase gene indicates a correlation between in vitro resistance to SP and the Asn-108 point mutation. Little is known about the association of this point mutation and in vivo resistance to SP. We used a mutation-specific polymerase chain reaction strategy to analyze the presence of the Asn-108 point mutation in 48 clinical samples with adequate clinical response (ACR), 2 early treatment failures (ETF), and 1 late treatment failure (LTF). The Asn-108 mutation was detected in 36 of the ACR samples and in all of the ETF and LTF samples. Eleven ACR samples amplified with the wild-type-specific primer and one amplified with the primer for the Thr-108 mutation described for resistance to cycloguanil. These results suggest that the Asn-108 marker may not be useful in predicting SP treatment failure. PMID- 10463675 TI - Nitric oxide, malaria, and anemia: inverse relationship between nitric oxide production and hemoglobin concentration in asymptomatic, malaria-exposed children. AB - The cause of the anemia associated with chronic, intermittent, asymptomatic, low level parasitemia in children in malaria-endemic endemic areas is not well understood. Nitric oxide (NO) decreases erythropoiesis, and it is likely an important mediator of anemia of chronic disease. Production of NO is decreased in acute uncomplicated and cerebral malaria, but it is increased in asymptomatic Tanzanian children (with or without parasitemia). We hypothesized that chronic overproduction of NO in these asymptomatic children contributes to the anemia associated with subclinical/subpatent malaria. In 44 fasting, asymptomatic, malaria-exposed, Tanzanian children, NO production (measured using fasting urine NOx excretion) was inversely associated with hemoglobin concentration (P = 0.03, controlling for age and gender). Using multiple linear regression, hemoglobin concentration was negatively associated with parasitemia (P = 0.005). After controlling for age and parasitemia, NO was no longer an independent predictor of anemia. One of the mechanisms of parasite-related anemia in such children may be through the adverse hematologic effects of parasite-induced NO production. PMID- 10463676 TI - Effects of age and parasitemia on nitric oxide production/leukocyte nitric oxide synthase type 2 expression in asymptomatic, malaria-exposed children. AB - Age appears to influence not only the acquisition of clinical immunity to malaria but also the susceptibility to and clinical manifestations of severe malaria. Asymptomatic malaria-exposed Tanzanian children have high production of nitric oxide (NO) and universal expression of leukocyte NO synthase type 2 (NOS2), which may protect against disease. To determine the effects of age and parasitemia on NO production, we measured urine and plasma NO metabolites and leukocyte NOS2 expression in 45 fasting, asymptomatic, malaria-exposed children of different ages, stratifying parasitemia by thick film and polymerase chain reaction (PCR) analysis. Although NO production was significantly higher in thick film-positive children than in thick film-negative children, after adjusting for age and gender, we were unable to detect a difference in NO production in thick film negative children between those who were PCR positive and PCR negative. The relationship between age and NO production was determined using a generalized additive model adjusted for the effects of gender and parasitemia. Production of NO using all three measures was highest in infancy, decreasing after the first year of life, and then increasing again after 5 years of age. This pattern of age related NO production is the reverse of the pattern of age-related morbidity from cerebral malaria in coastal Tanzanian children. Elevated production of NO in both infants and older children may be related to age per se and malaria infection respectively, and may be one of the mediators of the anti-disease immunity found most commonly in these two age groups. PMID- 10463678 TI - Comparative study on the ability of IgG and F(ab')2 antivenoms to neutralize lethal and myotoxic effects induced by Micrurus nigrocinctus (coral snake) venom. AB - A comparative study was performed on the ability of IgG and F(ab')2 antivenoms to neutralize lethal and myotoxic activities of Micrurus nigrocinctus venom. Both antivenoms were adjusted to a similar neutralizing potency in experiments where venom and antivenoms were preincubated prior to injection. No significant differences were observed between IgG and F(ab')2 antivenoms concerning neutralization of lethal effect in rescue experiments, i.e., when antivenom was administered intravenously after envenomation. However, F(ab')2 antivenom was more effective in prolonging the time of death when subneutralizing doses were administered immediately after venom injection. Both products partially reversed the binding of M. nigrocinctus alpha-neurotoxins to acetylcholine receptor in vitro. The IgG and F(ab')2 antivenoms effectively neutralized venom-induced myotoxicity when administered intravenously immediately after envenomation, although neutralization was poor if antivenom injections were delayed. Intramuscular injection of venom promoted diffusion of antivenom antibodies throughout muscle tissue, and F(ab')2 diffused to a higher extent than IgG molecules. Thus, despite the observation that F(ab')2 antivenom was more effective than IgG antivenom in prolonging the time of death when subneutralizing doses were administered immediately after envenomation, no major differences were observed in antivenom neutralization of lethal and myotoxic effects or in their capacity to reverse neurotoxin binding to the acetylcholine receptor. PMID- 10463677 TI - A new monospecific ovine Fab fragment antivenom for treatment of envenoming by the Sri Lankan Russell's viper (Daboia Russelii Russelii): a preliminary dose finding and pharmacokinetic study. AB - Russell's viper is the most important cause of life-threatening snake bite and acute renal failure in Sri Lanka. Only equine polyspecific antivenoms imported from India are available. They have not proved effective clinically or in clearing venom antigenemia and they frequently cause reactions. In an attempt to reduce mortality and morbidity, a new monospecific ovine Fab fragment antivenom (PolongaTab; Therapeutic Antibodies, Inc., London, United Kingdom) was raised against Sri Lankan Russell's viper venom. In a preliminary dose-finding study in 35 patients, an initial dose of 3-4 g restored blood coagulability permanently and stopped systemic bleeding, even in severely envenomed patients. Venom antigenemia disappeared within 1 hr of antivenom treatment but recurred, probably as a result of continued absorption of venom from the site of the bite, after the rapid clearance of therapeutic antibody. Twelve patients (34%) experienced early reactions that were usually mild and always responded to epinephrine. PMID- 10463679 TI - Macrophage responses to Toxoplasma antigens in vitro: a possible role in inflammatory lesions in toxoplasmosis. AB - Toxoplasma antigen and Toxoplasma immune complex were shown to induce increased production and release of acid hydrolases from macrophage cell line P388D in a concentration-dependent manner. Antigen concentrations of 10-50 microg/ml gave a 2-4-fold increase in the activities of acid proteinase, acid phosphatase, and phospholipase A2 compared with control cells without antigen. Results were similar for immune complex concentrations of 30-80 microg/ml compared with controls. No significant lactate dehydrogenase activity was detected in the culture medium, indicating that enzyme release was selective and not due to cell death. These results suggest that increased release of acid hydrolases may play a role in the inflammatory lesions observed in Toxoplasma encephalitis. PMID- 10463680 TI - Macrophage expression of class II major histocompatibility complex gene products in Paracoccidioides brasiliensis-infected mice. AB - C57B1/6 isogenic mice infected with Paracoccidioides brasiliensis strains showed a disruption in the expression of Ia antigen. Expression slowly decreased during the course of the infection with a slight variation dependent on the route of inoculation and the fungal strain used, but production of interferon-gamma and tumor necrosis factor-alpha were observed. Suppression of Ia antigen expression and depression of the immunoproliferative responses of spleen cells were strongly correlated with nitric oxide levels. These parameters were inhibited when the animals were treated with nitro-L-arginine, which resulted in inhibition the activation of nitric oxide (NO) production. Analysis of the data showed that changes in the expression of the Ia antigen occur in P. brasiliensis infection and are strongly correlated with NO levels. These phenomena may be interrelated and reflect macrophage activation that contributes to the control of the disease and to the immunosuppression observed during the course of the infection. PMID- 10463681 TI - Japanese encephalitis vaccine (inactivated, BIKEN) in U.S. soldiers: immunogenicity and safety of vaccine administered in two dosing regimens. AB - The safety and immunogenicity of Japanese encephalitis (JE) vaccine (Nakayama strain, monovalent / BIKEN) was studied in 538 U.S. soldiers in 1990. Three doses of vaccine from three consecutively manufactured lots were given on days 0, 7, and either 14 or 30. Serum for antibody determination was drawn at months 0, 2, and 6. Japanese encephalitis plaque reduction neutralization tests were performed by three laboratories on each specimen. Five hundred twenty-eight (98%) participants completed the immunization series. All recipients without antibody before immunization developed neutralizing antibody against JE virus. There were no differences in geometric mean titer among the three test lots at months 2 and 6. Soldiers who received the third dose on day 30 had higher titers at both time points. Antibody to yellow fever had no significant effect on immune response to vaccine. Conclusions drawn from analysis of serologic data from the three labs were nearly identical. Symptoms were generally limited to mild local effects and were reduced in frequency with each subsequent does in the series (21% to 11%; P < 0.0001). Generalized symptoms were rare (e.g., fever = 5%) with no reported cases of anaphylaxis. PMID- 10463682 TI - Short report: evaluation of the potency and stability of a candidate vaccine against American cutaneous leishmaniasis. AB - Availability of a safe, immunogenic, and affordable vaccine would represent the best strategy for control of cutaneous leishmaniasis (CL). Stability in field conditions is a essential property for any candidate vaccine. The stability and immunogenicity of three different preparations (thimerosal-preserved, autoclaved, and lyophilized) of a killed Leishmania amazonensis vaccine were assessed using fresh products and after 12 months of storage at 4 degrees C. Autoclaving was associated with a time-dependent decrease in the immunogenicity of the vaccine, as measured by the leishmanin skin test and production of interferon-gamma. These findings are of importance in the decision of which preparation of candidate killed CL vaccines should move to phase III trials. PMID- 10463683 TI - The fucose-mannose ligand-ELISA in the diagnosis and prognosis of canine visceral leishmaniasis in Brazil. AB - The fucose-mannose ligand (FML)-ELISA assay showed a sensitivity of 100% and a specificity of 100% in diagnosis of canine visceral leishmaniasis (CVL) (kala azar) in sera from naturally infected dogs from Sao Goncalo do Amaranto, Rio Grande de Norte, Brazil. The overall prevalence of antibodies to Leishmania in the endemic area was 23% (79 of 343). Seroreactivity detected by a Leishmania chagasi immunofluorescent (IF) assay was much lower (2.9%) and similar to the percentage of dogs with kala-azar symptoms (2.6%). Twenty-one of 21 asymptomatic, FML-seropositive animals died of kala-azar in a period ranging from 0 to 6 months after diagnosis. The predictive value was 100% for the FML-ELISA, 43% for an L. mexicana ELISA, and 24% for the L. mexicana and L. chagasi IF assays, respectively. In experimentally infected dogs, all assays detected seropositivity between 90 and 120 days after infection. Since the current strategy for control of CVL is based on detection and destruction of infected dogs, the highly predictive, sensitive, and specific FML-ELISA represents a useful tool for field control of the disease. PMID- 10463684 TI - Schistosomiasis mansoni: immunoblot analysis to diagnose and differentiate recent and chronic infection. AB - One hundred seven patients classified into three different groups (11 with acute schistosomiasis, 58 with chronic schistosomiasis, and 38 children with high IgM specific antibody titers against schistosome gut-associated antigens living in an endemic schistosomiasis area) were studied by immunoblotting for the presence of IgG, IgM, and IgA antibodies against Schistosoma mansoni soluble adult worm antigen preparation. We used sera from 15 individuals infected with various intestinal parasites, as well as sera from 19 uninfected individuals, as controls. An immunogenic fraction with a molecular weight of 31-32 kD (Sm31/32) was the most frequently recognized by the different antibody isotypes. In the group with acute disease, this fraction was recognized by IgG and IgM antibodies of all patients, and by 10 (90.9%) of 11 samples for IgA antibodies. Approximately 98% of the patients with chronic infections had IgG antibodies against Sm31/32, but only about 10% had IgM and IgA antibodies against this fraction. The IgG immunoblot profiles of the children from the endemic area were similar to those obtained for the group with acute schistosomiasis. This observation suggests recent infection of these children. Our data show that the Sm31/32 protein fraction is highly immunogenic and may be a useful serologic marker for diagnosing and differentiating between acute and chronic schistosomiasis infection. PMID- 10463685 TI - Early, intermediate, and late acute stages in Chagas' disease: a study combining anti-galactose IgG, specific serodiagnosis, and polymerase chain reaction analysis. AB - The acute phase of Chagas' disease was classified as early, intermediate, and late based on the levels of anti-Galalpha, 3Gal IgG (Gal) and specific IgM (M) and IgG (G) anti-T. cruzi reactivity. While the early phase was M+G-Gal-, the intermediate phase was M+G-Gal+, M+G+Gal-, or M+G+Gal+, and the late phase was M G+Gal+. This sequence of stages was consistent with our previous studies on acute phase proteins. Analysis by the polymerase chain reaction (PCR) of parasite DNA in 65 blood samples of children living in Cochabamba, Bolivia showed a significant correlation (90.8%) between ELISA and PCR positivity. A lower correlation was observed between indirect hemagglutination, PCR (58%), and ELISA. Electrocardiographic analysis of 43 children studied by the PCR did not show any alteration typical of acute chagasic myocarditis. The PCR positivity was observed in eight samples where only Gal was increased, suggesting a very early T. cruzi infection, when specific antibodies were not yet present. By associating anti-Gal IgG with specific serology, early T. cruzi infection can be detected with greater precision. We suggest the use of anti-Gal antibody reactivity as an aid for the detection of recent T. cruzi infections, at least in endemic areas where diseases caused by other trypanosomatids do not overlap. PMID- 10463686 TI - Resurgence of sleeping sickness in Tambura County, Sudan. AB - Endemic foci of human African trypanosomiasis are present in southern Sudan. In 1996 and 1997, trypanosomiasis increased sharply in Tambura County. To define the magnitude and geographic distribution of the outbreak, we conducted a prevalence survey using population-based cluster sampling in 16 villages: 1,358 participants answered questions about routine activities and tsetse fly contact and received serologic testing. Seroprevalence in the surveyed area was 19.4% (95% confidence interval = 16.9%, 21.8%). We confirmed infection in 66% of seropositive persons who received one parasitologic examination and in 95% of those who had serial examinations of lymph node fluid and blood. Activities related to the civil war, such as temporary migration, were not associated with seropositive status. Since the previous population screening in 1988, the trypanosomiasis prevalence increased two orders of magnitude, and the proportion of villages affected increased from 54% to 100%. Our results suggest that there may be 5,000 cases in Tambura County. The absence of trypanosomiasis control for nearly a decade is a factor in the resurgence of the disease. PMID- 10463687 TI - Acute disease episodes in a Wuchereria bancrofti-endemic area of Papua New Guinea. AB - Acute disease episodes of Bancroftian filariasis were monitored prospectively in a rural area of Papua New Guinea. The frequency and duration of episodes were recorded for the leg, arm, scrotum, and breast. A very high incidence of acute disease was observed; 0.31 episodes per person-year in the leg alone. Incidence generally increased with age, except in the breast, where episodes were concentrated in the reproductive age range. Males had slightly higher incidence than females in the leg and arm. Chronic disease was strongly associated with acute disease incidence in all locations. Microfilaremia had a statistically significant association with acute disease in the leg, arm, and breast, but not the scrotum. This study again demonstrates the high burden of acute manifestations of lymphatic filariasis, and provides new information on risk factors, which may lead to better understanding of etiology and control prospects. PMID- 10463688 TI - Natural rodent host associations of Guanarito and pirital viruses (Family Arenaviridae) in central Venezuela. AB - The objective of this study was to elucidate the natural rodent host relationships of Guanarito and Pirital viruses (family Arenaviridae) in the plains of central Venezuela. Ninety-two arenavirus isolates from 607 animals, representing 10 different rodent species, were characterized to the level of serotype. The 92 isolates comprised 19 Guanarito virus strains and 73 Pirital virus strains. The 19 Guanarito virus isolates were from Zygodontomys brevicauda; 72 (98.6%) of the 73 Pirital virus isolates were from Sigmodon alstoni. These results indicate that the natural rodent associations of these 2 sympatric arenaviruses are highly specific and that Z brevicauda and S. alstoni are the principal rodent hosts of Guanarito and Pirital viruses, respectively. PMID- 10463689 TI - Evidence for widespread infection of wild rats with hepatitis E virus in the United States. AB - Hepatitis E is an important medical pathogen in many developing countries but is rarely reported from the United States, although antibody to hepatitis E virus (anti-HEV) is found in > 1% of U.S. citizens. Zoonotic spread of the virus is suspected. Sera obtained from 239 wild rats trapped in widely separated regions of the United States were tested for anti-HEV. Seventy-seven percent of rats from Maryland, 90% from Hawaii, and 44% from Louisiana were seropositive for anti-HEV. Rats from urban as well as rural areas were seropositive and the prevalence of anti-HEV IgG increased in parallel with the estimated age of the rats, leading to speculation that they might be involved in the puzzling high prevalence of anti HEV among some U.S. city dwellers. The discovery of a in rats in the United States and the recently reported discovery that HEV is endemic in U.S. swine raise many questions about transmission, reservoirs, and strains of HEV in developed countries. PMID- 10463690 TI - Malaria parasitemia and childhood diarrhea in a peri-urban area of Guinea-Bissau. AB - To examine the association between diarrhea in early childhood and malaria parasitemia, we conducted a nested case-control study in Guinea-Bissau of 297 children with diarrhea and a similar number of children without diarrhea matched for age, season, and residential area. There were no associations between diarrhea and parasite rate, parasite density, or clinical malaria. However, anti malarials were easily available and frequently used, which was reflected by a 0.7% prevalence of children with a parasite density > 100/200 leukocytes. Thus, the findings do not preclude that diarrhea may be a sign of clinical malaria or high-parasite density in endemic areas with lower use of antimalarials. PMID- 10463691 TI - High rate of mixed and subpatent malarial infections in southwest Nigeria. AB - The rate of malarial parasitemia in children and adults was assessed by microscopy and the polymerase chain reaction in a holoendemic area in Nigeria. A high rate of subpatent Plasmodium falciparum parasitemia (19.6%) was found. Plasmodium malariae and P. ovale infections were common in a rural area (26.1% and 14.8%) but were observed sporadically in individuals from an urban area. Simultaneous infections with P. falciparum, P. malariae, and P. ovale were frequent in the rural area (11.7% triple infections). The rate of triple infections was higher than expected from the prevalences of each species (P < 0.00001). Spleen enlargement was associated with mixed infections of P. falciparum and P. malariae (odds ratio [OR] = 5.9, 95% confidence interval [CI] 3.0-11.7) and less frequently observed in individuals without detectable parasitemia (OR = 0.06, 95% CI = 0.01-0.3). Spleen enlargement and titers of antibodies to schizonts were positively correlated with parasite densities. The results also suggest that in some individuals a long-lasting subpatent parasitemia might occur. PMID- 10463692 TI - An outbreak of acute bartonellosis (Oroya fever) in the Urubamba region of Peru, 1998. AB - During May 1998, we conducted a case-control study of 357 participants from 60 households during an outbreak of acute bartonellosis in the Urubamba Valley, Peru, a region not previously considered endemic for this disease. Blood and insect specimens were collected and environmental assessments were done. Case patients (n = 22) were defined by fever, anemia, and intra-erythrocytic coccobacilli seen in thin smears. Most case-patients were children (median age = 6.5 years). Case-patients more frequently reported sand fly bites than individuals of neighboring households (odds ratio [OR] = 5.8, 95% confidence interval [CI] = 1.2-39.2), or members from randomly selected households > or = 5 km away (OR = 8.5, 95% CI = 1.7-57.9). Bartonella bacilliformis isolated from blood was confirmed by nucleotide sequencing (citrate synthase [g/tA], 338 basepairs). Using bacterial isolation (n = 141) as the standard, sensitivity, specificity, and positive predictive value of thin smears were 36%, 96%, and 44%, respectively. Patients with clinical syndromes compatible with bartonellosis should be treated with appropriate antibiotics regardless of thin-smear results. PMID- 10463694 TI - A piece of my mind. Stroke. PMID- 10463693 TI - Evidence of rickettsial spotted fever and ehrlichial infections in a subtropical territory of Jujuy, Argentina. AB - Between November 1993 and March 1994, a cluster 6 pediatric patients with acute febrile illnesses associated with rashes was identified in Jujuy Province, Argentina. Immunohistochemical staining of tissues confirmed spotted fever group rickettsial infection in a patient with fatal disease, and testing of serum of a patient convalescing from the illness by using an indirect immunofluorescence assay (IFA) demonstrated antibodies reactive with spotted fever group rickettsiae. A serosurvey was conducted among 16 households in proximity to the index case. Of 105 healthy subjects evaluated by IFA, 19 (18%) demonstrated antibodies reactive with rickettsiae or ehrlichiae: 4 had antibodies reactive with Rickettsia rickettsii, 15 with Ehrlichia chaffeensis, and 1 with R. typhi. Amblyomma cajennense, a known vector of R. rickettsii in South America, was collected from pets and horses in the area. These results are the first to document rickettsial spotted fever and ehrlichial infections in Argentina. PMID- 10463695 TI - Emergency departments open new doors to technology, patient service. PMID- 10463696 TI - Hormonally active agents throughout the environment. PMID- 10463697 TI - From the Centers for Disease Control and Prevention. Decline in deaths from heart disease and stroke--United States, 1900-1999. PMID- 10463698 TI - From the Centers for Disease Control and Prevention. Outbreak of Salmonella serotype Muenchen infections associated with unpasteurized orange juice--United States and Canada, June 1999. PMID- 10463699 TI - Doctor, you've got E-mail. PMID- 10463700 TI - Doctor, you've got E-mail. PMID- 10463701 TI - Doctor, you've got E-mail. PMID- 10463702 TI - Aspirin and risk of hemorrhagic stroke. PMID- 10463703 TI - Aspirin and risk of hemorrhagic stroke. PMID- 10463704 TI - Aspirin and risk of hemorrhagic stroke. PMID- 10463705 TI - Women's beliefs about breastfeeding and transmission of AIDS in rural Cote d'Ivoire. PMID- 10463706 TI - Association of the CCR5delta32 mutation with improved response to antiretroviral therapy. PMID- 10463707 TI - Utility of telemedicine on Johnston Atoll. PMID- 10463708 TI - A patient decision aid regarding antithrombotic therapy for stroke prevention in atrial fibrillation: a randomized controlled trial. AB - CONTEXT: Decision aids are tools designed to help patients participate in the clinical decision-making process. OBJECTIVE: To determine whether use of an audiobooklet (AB) decision aid explaining the results of a clinical trial affected the decision-making process of study participants. DESIGN: Randomized controlled trial conducted from May 1997 to April 1998. SETTING: Fourteen centers that participated in the Stroke Prevention in Atrial Fibrillation (SPAF) III trial. PARTICIPANTS: A total of 287 patients from the SPAF III aspirin cohort study, in which patients with atrial fibrillation and a relatively low risk of stroke received 325 mg/d of aspirin and were followed up for a mean of 2 years. INTERVENTION: At the end of SPAF III, participants were randomized to be informed of the study results with usual care plus use of an AB (AB group) vs usual care alone (control group). The AB included pertinent information to help patients decide whether to continue taking aspirin or switch to warfarin. MAIN OUTCOME MEASURES: Patients' ability to make choices regarding antithrombotic therapy, and 6-month adherence to these decisions. Their knowledge, expectations, decisional conflict (the amount of uncertainty about the course of action to take), and satisfaction with the decision-making process were also measured. RESULTS: More patients in the AB group made a choice about antithrombotic therapy than in the control group (99% vs 94%; P = .02). Patients in the AB group were more knowledgeable and had more realistic expectations about the risk of stroke and hemorrhage (in the AB group, 53%-80% correctly estimated different risks; in the control group, 16%-28% gave correct estimates). Decisional conflict and satisfaction were similar for the 2 groups. After 6 months, a similar percentage of patients were still taking their initial choice of antithrombotic therapy (95% vs 93%; P = .44). CONCLUSIONS: For patients with atrial fibrillation who had participated in a major clinical trial, the use of an AB decision aid improved their understanding of the benefits and risks associated with different treatment options and helped them make definitive choices about which therapy to take. Further studies are necessary to evaluate the acceptability and impact of decision aids in other clinical settings. PMID- 10463709 TI - HIV transmission through breastfeeding: a study in Malawi. AB - CONTEXT: Understanding the risk of human immunodeficiency virus (HIV) transmission through breastfeeding is essential for advising HIV-infected mothers and formulating public health policy recommendations. OBJECTIVE: To measure the frequency, timing, and risk factors of HIV transmission through breast milk. DESIGN: Prospective cohort study conducted between 1994 and 1997, with follow-up of infants through 24 months of age. SETTING: Postnatal clinic of tertiary care hospital, Blantyre, Malawi. PARTICIPANTS: A total of 672 infants (HIV-negative at birth) born to HIV-infected women who had not received antiretroviral drugs during or after pregnancy. MAIN OUTCOME MEASURE: Incidence of HIV in breastfed infants by age and maternal and infant risk factors for HIV transmission, using proportional hazard models to derive risk ratios (RRs) and 95% confidence intervals (CIs). RESULTS: Forty-seven children became HIV-infected while breastfeeding but none after breastfeeding had stopped. The cumulative infection rate while breastfeeding, from month 1 to the end of months 5, 11,17, and 23, was 3.5%, 7.0%, 8.9%, and 10.3%, respectively. Incidence per month was 0.7% during age 1 to 5 months, 0.6% during age 6 to 11 months, and 0.3% during age 12 to 17 months (P = .01 for trend). The only factors significantly associated with low risk of postnatal HIV transmission in a multivariate model were high maternal parity (RR, 0.23; 95% CI, 0.09-0.56) and older maternal age (RR, 0.44; 95% CI, 0.23-0.84). CONCLUSIONS: Our data suggest that the risk of HIV infection is highest in the early months of breastfeeding, which should be considered in formulating breastfeeding policy recommendations. PMID- 10463710 TI - Hypoglycemia and the decision to drive a motor vehicle by persons with diabetes. AB - CONTEXT: Laboratory studies have shown impairments in driving performance among subjects with type 1 diabetes mellitus when their blood glucose (BG) level is between 2.6 and 3.6 mmol/L (47-65 mg/dL). However, to our knowledge, no data exist examining subjects' decisions to drive at various BG levels during their daily routine. OBJECTIVE: To examine type 1 diabetic subjects' decisions to drive during their daily routine based on perception of BG levels compared with actual measured BG levels. DESIGN AND SETTING: Two separate groups of patients were recruited 2 years apart from 4 academic medical centers. PARTICIPANTS: All subjects were adults with type 1 diabetes who were drivers and who performed at least 2 BG tests per day. Group 1 (initial) subjects (n = 65) had a mean (SD) age of 38.6 (8.9) years with a mean (SD) diabetes duration of 20.5 (10.6) years, were taking 38.8 (16.8) U/d of insulin, and had a mean (SD) glycosylated hemoglobin (HbA1) level of 10.0% (1.9%). Group 2 (replication) subjects (n = 93) were 35.8 (8.0) years old with a mean diabetes duration of 17.0 (10.6) years, were taking 40.0 (15.5) U/d of insulin, and had a mean (SD) HbA1 level of 8.5% (1.6%). Each subject used a handheld computer to record data on symptoms, cognitive function, insulin dosage, food, activity, estimated and actual BG levels, and whether he/she would drive. Data were entered 3 to 6 times per day for a total of 50 to 70 collections per subject during a 3- to 4-week period. MAIN OUTCOME MEASURES: Decisions to drive when subjects estimated their BG level to be less than 2.2 mmol/L (40 mg/dL), 2.2 to 2.8 mmol/L (40-50 mg/dL), 2.8 to 3.3 mmol/L (50-60 mg/dL), 3.3 to 3.9 mmol/L (60-70 mg/dL), 3.9 to 10 mmol/L (70-180 mg/dL), and more than 10 mmol/L (>180 mg/dL), and driving decisions when actual BG levels were in these ranges. RESULTS: Subjects stated they would drive 43% to 44% of the time when they estimated their BG level to be 3.3 to 3.9 mmol/L (60-70 mg/dL), and 38% to 47% of the time when their actual BG level was less than 2.2 mmol/L (40 mg/dL). Logistic regression analysis demonstrated that number of autonomic symptoms, degree of impairment on cognitive function tests, and BG level estimate predicted 76% to 80% of decisions to drive (P<.01 for all). Approximately 50% of subjects in each group decided to drive at least 50% of the time when their BG level was less than 3.9 mmol/L (70 mg/dL). CONCLUSIONS: Our data suggest that persons with type 1 diabetes may not judge correctly when their BG level is too low to permit safe driving and may consider driving with a low BG level even when they are aware of the low level. Health care professionals should counsel their patients about the risk of driving with hypoglycemia and the importance of measuring BG level before driving. PMID- 10463711 TI - Psychiatric disorders among survivors of the Oklahoma City bombing. AB - CONTEXT: Disasters expose unselected populations to traumatic events and can be used to study the mental health effects. The Oklahoma City, Okla, bombing is particularly significant for the study of mental health sequelae of trauma because its extreme magnitude and scope have been predicted to render profound psychiatric effects on survivors. OBJECTIVE: To measure the psychiatric impact of the bombing of the Alfred P. Murrah Federal Building in Oklahoma City on survivors of the direct blast, specifically examining rates of posttraumatic stress disorder (PTSD), diagnostic comorbidity, functional impairment, and predictors of postdisaster psychopathology. DESIGN, SETTING, AND PARTICIPANTS: Of 255 eligible adult survivors selected from a confidential registry, 182 (71%) were assessed systematically by interviews approximately 6 months after the disaster, between August and December 1995. MAIN OUTCOME MEASURES: Diagnosis of 8 psychiatric disorders, demographic data, level of functioning, treatment, exposure to the event, involvement of family and friends, and physical injuries, as ascertained by the Diagnostic Interview Schedule/Disaster Supplement. RESULTS: Forty-five percent of the subjects had a postdisaster psychiatric disorder and 34.3% had PTSD. Predictors included disaster exposure, female sex (for any postdisaster diagnosis, 55% vs 34% for men; chi2 = 8.27; P=.004), and predisaster psychiatric disorder (for PTSD, 45% vs 26% for those without predisaster disorder; chi2 = 6.86; P=.009). Onset of PTSD was swift, with 76% reporting same day onset. The relatively uncommon avoidance and numbing symptoms virtually dictated the diagnosis of PTSD (94% meeting avoidance and numbing criteria had full PTSD diagnosis) and were further associated with psychiatric comorbidity, functional impairment, and treatment received. Intrusive reexperience and hyperarousal symptoms were nearly universal, but by themselves were generally unassociated with other psychopathology or impairment in functioning. CONCLUSIONS: Our data suggest that a focus on avoidance and numbing symptoms could have provided an effective screening procedure for PTSD and could have identified most psychiatric cases early in the acute postdisaster period. Psychiatric comorbidity further identified those with functional disability and treatment need. The nearly universal yet distressing intrusive reexperience and hyperarousal symptoms in the majority of nonpsychiatrically ill persons may be addressed by nonmedical interventions of reassurance and support. PMID- 10463712 TI - A 69-year-old man with anger and angina. PMID- 10463713 TI - A 45-year-old woman with premenstrual dysphoric disorder, 1 year later. PMID- 10463714 TI - Users' guides to the medical literature: XIX. Applying clinical trial results. A. How to use an article measuring the effect of an intervention on surrogate end points. Evidence-Based Medicine Working Group. PMID- 10463715 TI - The potential benefits of decision aids in clinical medicine. PMID- 10463716 TI - When is breastfeeding not best? The dilemma facing HIV-infected women in resource poor settings. PMID- 10463717 TI - Editors and owners--stretching reputation too far. PMID- 10463718 TI - Surrogate end points, health outcomes, and the drug-approval process for the treatment of risk factors for cardiovascular disease. PMID- 10463719 TI - Are surrogate markers adequate to assess cardiovascular disease drugs? PMID- 10463720 TI - JAMA Patient Page: diabetes. PMID- 10463721 TI - Hip replacement audit. PMID- 10463722 TI - Does thrombophilia cause Perthes' disease in children? PMID- 10463723 TI - Management of herniation of the lumbar disc. PMID- 10463724 TI - Outcome of Charnley total hip replacement across a single health region in England: the results at five years from a regional hip register. AB - Using a regional arthroplasty register we assessed the outcome at five years of 1198 primary Charnley total hip replacements (THRs) carried out in 1152 patients across a single UK health region in 1990. Information regarding outcome was available for 1080 hips (90%) and 499 had an independent clinical and radiological assessment. By five years the known rate of aseptic loosening was 2.3%, of deep infection 1.4%, of dislocation 5.0% and of revision 3.2%. The radiological assessment of 499 THRs revealed gross failure in a further 5.2%, which had been previously unrecognised. The combined rate of failure of nearly 9% is higher than those published from specialist centres and surgeons, but is probably more representative of the norm. Our study supports the need for a national register and surveillance of THRs. It emphasises that all implants should be followed, and suggests that the results of such surgery, when performed in the general setting, may not be as good as expected. PMID- 10463725 TI - Polyethylene wear, osteolysis and acetabular loosening with an HA-coated hip prosthesis. A follow-up of 94 consecutive arthroplasties. AB - We have followed up for a period of seven to nine years 100 consecutive arthroplasties of the hip in which an entirely HA-coated implant had been used. The clinical results were excellent and bony incorporation was extensive in all components. No stem became loose or subsided but five cups were revised because of loosening after 3.8 to 5.5 years, having functioned painlessly and shown radiological ingrowth. Revision procedures because of excessive polyethylene wear have been performed on 18 hips and are planned for six more. Two eroded metal backings with worn-through polyethylene were exchanged; six hips showed metallosis without polyethylene wear-through. There were two cases of granulomatous cysts in the groin and 66 hips had osteolysis located periarticularly, in the greater trochanter or in the acetabulum. PMID- 10463726 TI - Diagnostic criteria for non-traumatic osteonecrosis of the femoral head. A multicentre study. AB - Six major and seven minor diagnostic criteria have been developed by the Japanese Investigation Committee for osteonecrosis of the femoral head (ONFH). We have carried out a multicentre study to clarify these. We studied prospectively 277 hips in 222 patients, from six hospitals, who had ONFH and other hip pathology and from whom histological material was available. We identified five criteria with high specificity: 1) collapse of the femoral head without narrowing of the joint space or acetabular abnormality on radiographs, including the crescent sign; 2) demarcating sclerosis in the femoral head without narrowing or acetabular abnormality; 3) a 'cold-in-hot' appearance on the bone scan; 4) a low intensity band on T1-weighted images (band pattern); and 5) evidence of trabecular and marrow necrosis on histological examination. With any combination of two of these criteria, the sensitivity and specificity of the diagnosis were 91% and 99%, respectively. PMID- 10463727 TI - Acute fracture of the neck of the femur. An assessment of perfusion of the head by dynamic MRI. AB - We performed dynamic MRI of the femoral head within 48 hours of injury on 22 patients with subcapital fracture of the neck of the femur and on a control group of 20 of whom ten were healthy subjects and ten were patients with an intertrochanteric fracture. Three MRI patterns emerged when the results between the fractured side and the contralateral femoral head were compared. In all of the control group and in those patients who had undisplaced fractures (Garden stages I and II), perfusion of the femoral head was considered to be at the same level as on the unaffected side. In patients with displaced fractures (Garden stages III and IV) almost all the femoral heads on the fractured side were impaired or totally avascular, although some had the same level of perfusion as the unaffected side. We conclude that dynamic MRI, a new non-invasive imaging technique, is useful for evaluating the perfusion of the femoral head. PMID- 10463728 TI - Is diagnostic arthroscopy of the hip worthwhile? A prospective review of 328 adults investigated for hip pain. AB - We describe a prospective study of 328 patients undergoing arthroscopy of the hip. Arthrography, CT or MRI was performed when clinically indicated. A preoperative diagnosis was reached in 174 patients (53%), while the remaining 154 were diagnosed as having 'idiopathic hip pain'. In seven patients, access to the hip was inadequate. Arthroscopy altered the diagnosis in 176 hips (53%). The new primary diagnoses were osteoarthritis (75 patients), osteochondral defects (34), torn labra (23), synovitis (11) and loose bodies (9). In 172 hips (52%) an operative procedure was undertaken. In the remaining 84 patients (26%), arthroscopy neither changed the diagnosis nor provided surgical treatment. Arthroscopy of the hip is considered to be of value in assessing and treating the adult patient with pain in the hip of uncertain cause. PMID- 10463729 TI - Complications of arthroscopy of the hip. AB - Although arthroscopy of the hip is being carried out increasingly, little is known about the rate of associated complications. We describe a prospective study of 640 consecutive procedures in which a consistent technique was used. The overall complication rate was 1.6%. Complications, none of which was major or long-term, included transient palsy of the sciatic and femoral nerves, perineal injury, bleeding from the portal wounds, trochanteric bursitis and intra articular breakage of the instrument. We believe that it is possible to undertake this operation safely using the technique described. PMID- 10463730 TI - Tarsal tunnel syndrome caused by ganglia. AB - We describe in 30 feet the occurrence of a tarsal tunnel syndrome caused by a ganglion. The presenting symptom was numbness or pain in the toes and the sole with paraesthesiae in the distribution of the medial plantar nerve in 63% of the patients. Swellings which were not palpable were detected by ultrasonography. Twenty-nine patients were treated by operation. Most ganglia originated from the talocalcaneal joint, and five were associated with a talocalcaneal coalition. The surgical outcome was satisfactory in all patients except one who had a further operation for a recurrence of the ganglion. PMID- 10463731 TI - Subtalar arthrodesis with correction of deformity after fractures of the os calcis. AB - We have reviewed the long-term results of 22 patients (23 fusions) with fractures of the os calcis, who had subtalar arthrodesis with correction of the deformity between 1975 and 1991. The mean follow-up was nine years (5 to 20). All patients were evaluated according to a modified foot score. A radiological assessment was used in which linear and angular variables were measured including the fibulocalcaneal abutment, the height of the heel and fat pad, the angle of the arch and the lateral talocalcaneal and the lateral talar declination angles. The technique used restores the normal relationship between the hindfoot and midfoot and corrects the height of the heel. This leads to better biomechanical balance of the neighbouring joints and gives a favourable clinical outcome. The modified foot score showed a good or excellent result in 51% of the feet. Residual complaints were mostly due to problems with the soft tissues. Subjectively, an excellent or good score was achieved in 78% of the cases. After statistical analysis, except for the height of the heel and the degenerative changes in the calcaneocuboid joint, no significant difference was found in the measured variables between the operated and the contralateral side. PMID- 10463732 TI - Disarticulation at the ankle using an anterior flap. A preliminary report. AB - Disarticulation has been carried out in ten ankles in nine patients in whom it was not possible to use a heel flap. Four patients were able to walk with a prosthesis which gave satisfactory function. In five who were bedridden, healing was achieved and was of sufficient quality to allow transfers. There was no operative morbidity or mortality. This technique can be used instead of a transtibial amputation if necrosis or ischaemia of the heel is a contraindication to conventional Syme's amputation. PMID- 10463733 TI - Screw versus suture fixation of Mitchell's osteotomy. A prospective, randomised study. AB - We studied prospectively 30 patients who had a Mitchell's osteotomy secured by either a suture followed by immobilisation in a plaster boot for six weeks, or by a cortical screw with early mobilisation. The mean time for return to social activities after fixation by a screw was 2.9 weeks and to work 4.9 weeks, which was significantly earlier than those who had stabilisation by a suture (5.7 and 8.7 weeks, respectively; p < 0.001). Use of a screw also produced a higher degree of patient satisfaction at six weeks, and an earlier return to wearing normal footwear. The improvement in forefoot scores was significantly greater after fixation by a screw at six weeks (p = 0.036) and three months (p = 0.024). At one year, two screws had been removed because of pain at the site of the screw head. Internal fixation of Mitchell's osteotomy by a screw allows the safe early mobilisation of patients and reduces the time required for convalescence. PMID- 10463734 TI - The knee joint in diastrophic dysplasia. A clinical and radiological study. AB - We examined clinically and radiologically the knees of 46 patients (27 females and 19 males) with diastrophic dysplasia. The age of the patients varied from newborn to 38 years. A total of 18 patients was followed during their growth until adolescence. The knees of two legally aborted fetuses appeared on examination to be macroscopically normal and congruous. Excessive valgus deformity of the tibiofemoral weight-bearing angle with a mean of 14 degrees was noted in infancy. Most of the patients had marked instability of the knees. The range of movement of the knee began to decrease before the age of five years. There were signs of early degeneration and deformation of the bony epiphyses before the age of six years. The patellofemoral joint was abnormal from an early age. A marked patella infera, often associated with a lateral position of the patella with bony fragmentation, was noted. The knee in diastrophic dysplasia is basically unstable, showing early deformation of the subchondral bone and degeneration of the joint. PMID- 10463735 TI - Plain radiography in the degenerate knee. A case for change. AB - We took posteroanterior weight-bearing radiographs, both with the joint fully extended and in 30 degrees of flexion, in a consecutive series of 50 knees in 37 patients referred for the primary assessment of pain and/or stiffness. These radiographs were reported 'blind' both by an orthopaedic surgeon and a radiologist. Direct measurement of the joint space, together with grading of the severity of erosion according to the Ahlback criteria, was undertaken. Any other abnormality present was also documented. The radiographs of the knees in 30 degrees of flexion consistently showed more advanced erosion in both the medial (p = 0.001) and the lateral (p = 0.0001) tibiofemoral compartments, when compared with those of knees in full extension. The Ahlback classification of 25 joints was altered, in some cases by several grades, by the flexed position of the joint. In every case in which another abnormality was identified on the radiograph in full extension, it was also noted on that of the knee in 30 degrees of flexion. In a further four cases, additional pathology could only be seen in the flexed knee. Every patient was able to complete the radiological examination without difficulty. Our study supports the adoption of a weight-bearing view in 30 degrees of flexion as the standard posteroanterior radiograph for the assessment of tibiofemoral osteoarthritis in patients over 50 years of age. PMID- 10463736 TI - The popliteofibular ligament. An anatomical study of the posterolateral corner of the knee. AB - We designed an experimental study to prove the existence of the popliteofibular ligament (PFL) and to define its role in providing static stability of the knee. We also examined the contribution of the lateral collateral ligament (LCL). We found this ligament to be present in all eight human cadaver knees examined. These specimens were mounted on a specially designed rig and subjected to posterior, varus and external rotational forces. We used the technique of selective sectioning of ligaments and measured the displacement with a constant force applied, before and after its division. We recorded the displacement in primary posterior translation, coupled external rotation, primary varus angulation and primary external rotation. Statistical analysis using the standard error of the mean by plotting 95% confidence intervals, was used to evaluate the results. The PFL had a significant role in preventing excessive posterior translation and varus angulation, and in restricting excessive primary and coupled external rotation. Isolated section of the belly of popliteus did not cause significant posterolateral instability of the knee. The LCL was also seen to act as a primary restraint against varus angulation and secondary restraint against external rotation and posterior displacement. Our findings showed that in knees with isolated disruption of the PFL stability was restored when it was reconstructed. However in knees in which the LCL was also disrupted, isolated reconstruction of the PFL did not restore stability. PMID- 10463737 TI - Subperiosteal ganglion cyst of the tibia. A communication with the knee demonstrated by delayed arthrography. AB - We report a patient with a subperiosteal ganglion cyst of the tibia which was imaged by radiography, arthrography, CT and MRI. The images were correlated with the arthroscopic surgical and histological findings. Spiculated formation of periosteal new bone on plain radiographs led to the initial suspicion of a malignant tumour. Demonstration of the cystic nature of the tumour using cross sectional imaging was important for the precise diagnosis. Communication between the ganglion cyst and the knee was shown by a delayed arthrographic technique, and the presence of this communication was confirmed at arthroscopy and surgically. PMID- 10463738 TI - The results at ten years of the Insall-Burstein II total knee replacement. Clinical, radiological and survivorship studies. AB - We reviewed the outcome of 146 Insall-Burstein II total knee replacements carried out in 121 patients over a period of nearly four years in a general orthopaedic unit. At a mean follow-up of ten years, 94 knees in 78 patients were available for review. Six patients (7 knees) were lost to follow-up and 37 (45 knees) had died. The clinical outcome using the scoring system of the Hospital for Special Surgery (HSS) was excellent or good in 79% of patients, fair in 14% and poor in 7%. The mean preoperative HSS score was 31, improving to 79 at the latest review. Using the newer rating system of the Knee Society, the mean score at ten years was 87 and the mean functional score 56. The arc of flexion improved from a mean preoperative value of 88 degrees to 100 degrees. The 18 patients who had had a previous high tibial osteotomy were analysed separately and were found to have benefited equally from the operation. Nine prostheses were revised, giving a cumulative survival rate of 92.3% at ten years. Radiological evaluation of 104 radiographs showed radiolucent lines around ten tibial components, none of which required revision. Anterior knee pain was a significant problem. PMID- 10463739 TI - Prevention of deep-vein thrombosis after total knee replacement. Randomised comparison between a low-molecular-weight heparin (nadroparin) and mechanical prophylaxis with a foot-pump system. AB - The optimal regime of antithrombotic prophylaxis for patients undergoing total knee arthroplasty (TKA) has not been established. Many surgeons employ intermittent pneumatic compression while others use low-molecular-weight heparins (LMWH) which were primarily developed for total hip arthroplasty. We compared the efficacy and safety of these two techniques in a randomised study with blinded assessment of the endpoint by phlebography. We randomised 130 patients, scheduled for elective TKA, to receive one daily subcutaneous injection of nadroparin calcium (dosage adapted to body-weight) or continuous intermittent pneumatic compression of the foot by means of the arteriovenous impulse system. A total of 108 patients (60 in the LMWH group and 48 in the mechanical prophylaxis group) had phlebography eight to 12 days after surgery. Of the 47 with deep-vein thrombosis, 16 had received LMWH (26.7%, 95% CI 16.1 to 39.7) and 31, mechanical prophylaxis (64.6%, 95% CI 49.5 to 77.8). The difference between the two groups was highly significant (p < 0.001). Only one patient in the LMWH group had severe bleeding. We conclude that one daily subcutaneous injection of calcium nadroparin in a fixed, weight-adjusted dosage scheme is superior to intermittent pneumatic compression of the foot for thromboprophylaxis after TKA. The LMWH scheme was also safe. PMID- 10463740 TI - Health outcome after total knee replacement in the very elderly. AB - Between 1992 and 1994 we performed a prospective study of the effect of total knee replacement (TKR) on the health status of 119 patients over the age of 80 years who had had a primary unilateral TKR. The Nottingham Health Profile was used to assess this before and at three and 12 months after operation. We found a significant improvement in the scores for pain, emotional reaction, sleep and physical mobility at three months. After 12 months, the scores for pain and sleep were well maintained. The other factors had deteriorated slightly but remained better than before operation. Our findings show that TKR leads to a significant improvement in the general health status of the very elderly. PMID- 10463741 TI - Nonunion of tibial stress fractures in patients with deformed arthritic knees. Treatment using modular total knee arthroplasty. AB - In two years we treated four women with ununited stress fractures of their proximal tibial diaphyses. They all had arthritis and valgus deformity. The stress fractures had been treated elsewhere by non-operative means in three patients and by open reduction and internal fixation in one, but had failed to unite. After treatment with a modular total knee prosthesis with a long tibial stem extension, all the fractures united. A modular total knee prosthesis is suitable for the rare and difficult problem of ununited tibial stress fractures in patients with deformed arthritic knees since it corrects the deformity and the adverse biomechanics at the fracture site, stabilises the fracture and treats the arthritis. PMID- 10463742 TI - Osteochondroma with compression of the spinal cord. A report of two cases. AB - We report two cases of vertebral osteochondroma. In one patient a solitary cervical lesion presented as entrapment neuropathy of the ulnar nerve and in the other as a thoracic tumour associated with hereditary multiple exostoses producing paraplegia. We highlight the importance of an adequate preoperative evaluation in such patients. PMID- 10463743 TI - Degenerative spondylolisthesis. Developmental or acquired? AB - Degenerative spondylolisthesis is four times more common in women than in men. Although this gender difference has long been recognised there has been no explanation for it. We have examined the radiographs and CT scans of 118 patients over the age of 55 years and of a control group under the age of 46 years. Our findings confirmed the presence of more sagittally-orientated facet joints in patients with degenerative spondylolisthesis but did not show that the gender difference can be explained by the morphology of the facet joint. Furthermore, we conclude that the increased angle of the facet joint is the result of arthritic remodelling and not the primary cause of degenerative spondylolisthesis. It is more likely to be due to loss of soft-tissue resilience with subsequent failure of the facet joints which are acting as the last restraints to subluxation. PMID- 10463744 TI - Evaluation of chronic tears of the rotator cuff by ultrasound. A new index. AB - The diagnosis of chronic lesions of the rotator cuff is challenging. We have developed a new index to improve the sonographic diagnosis of chronic tears of the cuff. In a pilot study, we examined 50 asymptomatic healthy volunteers by ultrasound to establish the diameter of the rotator cuff in relation to the tendon of the long head of biceps. Subsequently, the index was calculated in 64 patients who had had shoulder pain for more than three months caused by clinically diagnosed lesions of the rotator cuff. The compensatory hypertrophy of the biceps tendon was quantified sonographically in relation to the diameter of the cuff. Comparison with the contralateral shoulder revealed a significantly higher biceps rotator-cuff ratio (p < 0.05) for patients with torn rotator cuffs. A ratio greater than 0.8 was considered pathological (index positive); the mean ratio in the control group was 0.43. The sensitivity of a positive index was 97.8%, the specificity 63.2%, the positive predictive value 86.3%, and the negative predictive value 92.4% in comparison with surgical findings. Use of the index improves sensitivity in the diagnosis of chronic tears of the cuff by ultrasound. PMID- 10463745 TI - The incidence of nerve injury in anterior dislocation of the shoulder and its influence on functional recovery. A prospective clinical and EMG study. AB - Opinion varies as to the incidence of nerve lesions in anterior dislocation of the shoulder after low-velocity trauma. Most studies are retrospective or do not use EMG. We have investigated the incidence and the clinical consequences of nerve lesions in a prospective study by clinical and electrophysiological examination. Axonal loss was seen in 48% of 77 patients. The axillary nerve was most frequently involved (42%). Although recovery as judged by EMG and muscle strength was almost complete, function of the shoulder was significantly impaired in patients with lesions of the axillary and suprascapular nerves. Unfavourable prognostic factors are increasing age and the presence of a haematoma. It is not necessary to carry out EMG routinely; an adequate programme of physiotherapy is important. In patients with a severe paresis, EMG is essential after three weeks. PMID- 10463746 TI - Does thrombophilia play an aetiological role in Legg-Calve-Perthes disease? AB - Heritable thrombophilic disorders have been proposed as one of the causes for Legg-Calve-Perthes disease. A total of 62 patients diagnosed with this disease between 1988 and 1997 and 50 controls were screened for thrombophilia. The incidence and relationship of thrombophilia to the severity of the disease were evaluated. One patient and none of the controls had protein S deficiency. One of the control group and one of the patients had protein C deficiency with the latter child also having a combined deficiency with a mutant factor V gene. The number of children with a mutant factor V gene, protein C deficiency, who were homozygous for the C 677T polymorphism of methylenetetrahydrofolate reductase or were heterozygous for mutant G20210A prothrombin did not differ statistically in the study and the control groups. No patient had antithrombin deficiency or positive lupus anticoagulant. We found no correlation between thrombophilia and the extent of the disease. The most common risk factors for arteriovenous thromboembolism showed no statistical significance in our patients compared with the control group or with the general population. These data do not confirm an aetiological role for thrombophilia in Perthes' disease. PMID- 10463747 TI - Perthes' disease and the relevance of thrombophilia. AB - Recent work has suggested that thrombophilia may be an aetiological factor in up to 50% of children with Perthes' disease, and that up to 75% may have a coagulopathy. Our aim was to test these findings in the local population of children with Perthes' disease and attempt to correlate them with the severity of the condition. In 64 children there were only eight (12%) with low levels of clotting proteins, as defined by normal paediatric ranges. Of these eight, only five could be said to show any thrombophilic tendency. PMID- 10463748 TI - Perthes' disease after the age of twelve years. Role of the remaining growth. AB - In order to define the prognostic factors in Perthes' disease in children older than 12 years, we reviewed 15 patients at the end of growth who were aged 12.1 to 14 years at presentation. The patients with the worst long-term prognosis (Stulberg class V) were compared with the others for age, skeletal maturity and remaining growth (Oxford method), as well as Catterall and Waldenstrom classifications at presentation. A significant difference (p = 0.001) was found for remaining growth (25% in Stulberg class V and 35% in the others) and also for the results at the end of growth when the remaining growth was over 30%, since this allowed sufficient time for reformation and remodelling of the femoral head. PMID- 10463749 TI - The relationship of foot and ankle movements to venous return in the lower limb. AB - We have studied the relationship between movements of the foot and ankle and venous blood flow from the lower limb using colourflow Duplex ultrasound to determine the optimum type of exercise for promoting venous return. Studies of both active and passive movements were carried out on 40 limbs in 20 subjects (18 men; 2 women), with a median age of 27 years (20 to 54). We assessed ankle dorsiflexion and plantar flexion, subtalar inversion and eversion, and a combination of all movements. There was no difference in venous flow when comparing opposite limbs in a single subject (p > 0.5), but active exercises produced higher peak and mean velocities of blood flow than passive ones. The active combined movement produced the highest velocities with an increase of 38% in mean and of 58% in peak flow velocities, which were significantly greater than the peak and mean flow rates produced by passive movements. The active combined exercise would therefore be the most effective in eliminating stasis and could contribute to the prevention of deep-vein thrombosis. PMID- 10463750 TI - The effect of hydroxyapatite coating on the bonding of bone to titanium implants in the femora of ovariectomised rats. AB - We have studied the effect of hydroxyapatite (HA) coating in 15 ovariectomised and 15 normal rats which had had a sham procedure. Twenty-four weeks after operation, HA-coated implants were inserted into the intramedullary canal of the right femur and uncoated implants into the left femur. The prostheses were removed four weeks after implantation. Twelve specimens in each group had mechanical push-out tests. Sagittal sections of the other three were evaluated by SEM. The bone mineral density (BMD) of the dissected left tibia was measured by dual-energy x-ray absorptiometry. The difference in BMD between the control and ovariectomised tibiae was 35.01 mg/cm2 (95% CI, 26.60 to 43.42). The push-out strength of the HA-coated implants was higher than that of the uncoated implants in both groups (p < 0.0001), but the HA-coated implants of the ovariectomised group had a reduction in push-out strength of 40.3% compared with the control group (p < 0.0001). Our findings suggest that HA-coated implants may improve the fixation of a cementless total hip prosthesis but that the presence of osteoporosis may limit the magnitude of this benefit. PMID- 10463751 TI - Osteogenic activity of OP-1 bone morphogenetic protein (BMP-7) in a human fibular defect. AB - We performed a prospective, randomised double-blind study in 24 patients undergoing high tibial osteotomy to evaluate the effectiveness of human recombinant osteogenic protein (OP-1) on a collagen type-I carrier in a critically-sized fibular defect. The study had two phases, each evaluated by clinical, radiological and DEXA methods during the first postoperative year. The first concerned the validation of the model of the fibular defect, using positive (demineralised bone) and negative (untreated) controls. The second phase concerned the osteogenic potential of OP-1 on collagen type-I v collagen type-I alone. The results of the first phase established the critically-sized nature of the defect. In the untreated group no bony changes were observed while, in the demineralised bone group, formation of new bone was visible from six weeks onwards. The results of the second phase showed no significant formation of new bone in the presence of collagen alone, while in the OP-1 group, all patients except one showed formation of new bone from six weeks onwards. This proved the osteogenic activity of OP-1 in a validated critically-sized human defect. PMID- 10463752 TI - Coral grafting supplemented with bone marrow. AB - Limited success in regenerating large bone defects has been achieved by bridging them with osteoconductive materials. These substitutes lack the osteogenic and osteoinductive properties of bone autograft. A direct approach would be to stimulate osteogenesis in these biomaterials by the addition of fresh bone-marrow cells (BMC). We therefore created osteoperiosteal gaps 2 cm wide in the ulna of adult rabbits and either bridged them with coral alone (CC), coral supplemented with BMC, or left them empty. Coral was chosen as a scaffold because of its good biocompatibility and resorbability. In osteoperiosteal gaps bridged with coral only, the coral was invaded chiefly by fibrous tissue. It was insufficient to produce union after two months. In defects filled with coral and BMC an increase in osteogenesis was observed and the bone surface area was significantly higher compared with defects filled with coral alone. Bony union occurred in six out of six defects filled with coral and BMC after two months. An increase in the resorption of coral was also observed, suggesting that resorbing cells or their progenitors were present in bone marrow and survived the grafting procedure. Our findings have shown that supplementation of coral with BMC increased both the resorption of material and osteogenesis in defects of a clinical significance. PMID- 10463753 TI - The influence of crystallinity of the hydroxyapatite coating on the fixation of implants. Mechanical and histomorphometric results. AB - We inserted two hydroxyapatite (HA)-coated implants with crystallinities of either 50% (HA-50%) or 75% (HA-75%) bilaterally into the medial femoral condyles of the knees of 16 dogs. The implants were allocated to two groups with implantation periods of 16 and 32 weeks. They were weight-bearing and subjected to controlled micromovement of 250 microm during each gait cycle. After 16 weeks, mechanical fixation of the HA-50% implants was increased threefold as compared with the HA-75% implants. After 32 weeks there was no difference between HA-50% and HA-75%. Fixation of HA-75% increased from 16 to 32 weeks whereas that of HA 50% was unchanged. HA-50% implants had 100% more bone ingrowth than HA-75% implants after 16 weeks. More HA coating was removed on HA-50% implants compared with HA-75% implants after both 16 and 32 weeks. No further loss of the HA coating was shown from 16 to 32 weeks. Our study suggests that the crystallinity of the HA coating is an important factor in its bioactivity and resorption during weight-bearing conditions. Our findings suggest two phases of coating resorption, an initial rapid loss, followed by a slow loss. Resorbed HA coating was partly replaced by bone ingrowth, suggesting that implant fixation will be durable. PMID- 10463754 TI - Dupuytren's disease. A model for the mechanism of fibrosis and its modulation by steroids. AB - Dupuytren's disease is a chronic inflammatory process which produces contractures of the fingers. The nodules present in Dupuytren's tissue contain inflammatory cells, mainly lymphocytes and macrophages. These express a common integrin known as VLA4. The corresponding binding ligands to VLA4 are vascular cell adhesion molecule-1 (VCAM-1) present on the endothelial cells and the CS1 sequence of the fibronectin present in the extracellular matrix. Transforming growth factor-beta (TGF-beta) is a peptide hormone which has a crucial role in the process of fibrosis. We studied tissue from 20 patients with Dupuytren's disease, four samples of normal palmar fascia from patients undergoing carpal tunnel decompression and tissue from ten patients who had received perinodular injections of depomedrone into the palm five days before operation. The distribution of VLA4, VCAM-1, CS1 fibronectin and TGF-beta was shown by immunohistochemistry using an alkaline phosphorylase method for light microscopy. In untreated Dupuytren's tissue CS1 fibronectin stained positively around the endothelial cells of blood vessels and also around the surrounding myofibroblasts, principally at the periphery of many of the active areas of the Dupuytren's nodule. VCAM-1 stained very positively for the endothelial cells of blood vessels surrounding and penetrating the areas of high nodular activity. VCAM-1 was more rarely expressed outside the blood vessels. VLA4 was expressed by inflammatory cells principally in and around the blood vessels expressing VCAM-1 and CS1 but also on some cells spreading into the nodule. TGF-beta stained positively around the inflammatory cells principally at the perivascular periphery of nodules. These cells often showed VLA4 expression and co-localised with areas of strong production of CS1 fibronectin. Normal palmar fascia contained only scanty amounts of CS1 fibronectin, almost no VCAM-1 and only an occasional cell staining positively for VLA4 or TGF-beta. In the steroid-treated group, VCAM-1 expression was downregulated in the endothelium of perinodular blood vessels and only occasional inflammatory cell expression remained. Expression of CS1 fibronectin was also much reduced but still occurred in the blood vessels and around the myofibroblast stroma. VLA4-expressing cells were also reduced in numbers. A similar but reduced distribution of production of TGF beta was also noted. Our findings show that adherence of inflammatory cells to the endothelial wall and the extravasation into the periphery of the nodule may be affected by steroids, which reduce expression of VCAM-1 in vivo. This indicates that therapeutic intervention to prevent the recommencement of the chronic inflammatory process and subsequent fibrosis necessitating further surgery may be possible. PMID- 10463755 TI - Reamed versus unreamed nails. PMID- 10463756 TI - Pain in the assessment of total knee replacement. PMID- 10463757 TI - Open acromioplasty does not prevent the progression of an impingement syndrome to a tear. PMID- 10463758 TI - Retroversion of the acetabulum. PMID- 10463759 TI - Neonatal detection of developmental dysplasia of the hip (DDH) PMID- 10463760 TI - Patterns of weight distribution under the metatarsal heads. PMID- 10463761 TI - Infections associated with dental procedures in total hip arthroplasty. PMID- 10463762 TI - Salvage of the head of the radius after fracture-dislocation of the elbow. PMID- 10463763 TI - Effects of sublethal radiation on bone marrow cells: induction of apoptosis and inhibition of antibody formation. AB - As part of the study to investigate the mechanism of radiation-induced immunosuppression, the survival and functional ability of bone marrow cells was analyzed by exposing C57B1/6 mice whole body to 2.0-Gy gamma-rays. There was a rapid induction of DNA fragmentation in the total bone marrow cells and the kinetics indicated that apoptosis reached a peak by 4 h and then dropped back to normal control levels within 10 h after irradiation. To determine the functional ability of bone marrow cells which survive the radiation treatment, animals were immunized with antigen trinitrophenyl (TNP)-lipopolysaccharide. There was a significant decrease of anti-TNP plaque-forming cells in the bone marrow of irradiated mice compared to control animals. Flow cytometric analysis of bone marrow revealed a significant depletion of both immature (B220+, Ig-) as well as mature (B220+, Ig+) B cells compared to control group. In summary, the present study showed that sublethal whole body irradiation inhibits antibody responses elicited by bone marrow cells. This decreased immune response may have been due to depletion of B lineage subsets as well as generalized apoptosis in the entire bone marrow cells. PMID- 10463764 TI - Lead intoxication: antioxidant defenses and oxidative damage in rat brain. AB - Oxidative damage associated with the presence of lead (Pb) in the brain has been proposed as one possible mechanism involved in Pb toxicity. To investigate this hypothesis, we examined the long-term effects of Pb2+ on parameters of oxidative stress in the brain from rats chronically exposed to the metal (1 g Pb acetate/1 drinking water). After 8 weeks of treatment, Pb2(+)-intoxicated rats (blood Pb concentration > 100 microg/dl) showed lower body weight, and lower hematocrit and 5-aminolevulinic acid dehydratase activity as compared to controls. The content of brain 2-thiobarbituric acid-reactive substances (TBARS), an indicator of lipid oxidation, was significantly (P < 0.05) higher in the Pb2(+)-intoxicated animals than in controls. Higher activities of the antioxidant enzymes glutathione reductase and glutathione peroxidase, and a lower (44%) level of ubiquinol 10 were found in the brain of the Pb2(+)-treated rats, compared to controls. A negative correlation between brain ubiquinol 9 (r2 = 0.79), 10 (r2 = 0.84) and blood Pb concentration was observed. Brain alpha-tocopherol levels, superoxide dismutase activity and parameters of oxidative damage to proteins were similar between control and Pb2(+)-treated rats. The present results indicate that chronic Pb2+ intoxication induces an oxidative stress situation in rat brain. PMID- 10463765 TI - Effect of inhalation exposure to 2-bromopropane on the nervous system in rats. AB - Exposure to 2-bromopropane (2-BP) is suspected to have adverse effects on the nervous system. The aim of this study was to investigate whether the exposure of rats to 2-BP had neurotoxic effects using histological and electrophysiological studies. Wistar strain male rats were exposed daily to either 100 or 1000 ppm 2 BP or to fresh air for 8 h a day for 12 weeks. Body weight was measured before exposure and every 2 weeks. Motor nerve conduction velocity (MCV) and distal latency (DL) were measured before exposure and every 4 weeks during exposure. Histological examination of the nervous system was also performed. Exposure of rats (n = 9) to 1000 ppm resulted in suppression of body weight gain and a significant decrease in brain weight compared to the control (n = 9). Electrophysiological measurements showed a significant decrease in MCV in 1000 ppm exposed rats at 8 weeks and significant prolongation of DL at 8 and 12 weeks. Abnormalities of the myelin sheath were detected in the common peroneal nerves. In 100-ppm exposed rats (n = 9), no significant changes were noted in body weight and the peripheral nerve. In conclusions, long-term exposure to 1000 ppm of 2-BP may result in peripheral neuropathy in rats. PMID- 10463766 TI - Differential toxicities of TCDD in vivo among normal, c-src knockout, geldanamycin- and quercetin-treated mice. AB - Although we have previously reported the result of our preliminary study on the reduced toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in vivo in male c-src deficient, -/+ B6, 129-src(tmlSor) mice as compared to those in wild type mice, there are still two major shortcomings of the above study: (a) the low number of individuals tested, (b) in some of the comparison tests, C57BL/6J mice (i.e. inbred B6 mice) were used as wild-type control mice. Since then we increased our laboratory breeding program and thereby the availability of B6, 129 /+, -/- and true littermate wild-type +/+ individual mice. The results of critical in vivo toxicity comparison tests, involving 6-13 mice per test group, showed that there are considerable variations expressed in toxicity within each group of c-src deficient mice. Nevertheless, when a large enough number of individuals were tested two toxic effects were found to be less expressed in src deficient mice. They were: (a) excess fatty deposits and (b) the mottled appearance of the liver which were commonly observed in TCDD-treated wild type mice, but not in c-src deficient mice. The former trend was also confirmed by both liver lipid analysis and histological examinations of the affected livers. As for the biochemical parameters, the hepatic nuclear protein binding to C/EBP (CCAAT/enhancer binding protein) response element appears to be uniformly reduced by the action of TCDD in + / + mice, but not in -/+ or -/- mice. PMID- 10463768 TI - Quantitative determination of apoptotic death in cultured human pancreatic cancer cells by propidium iodide and digitonin. AB - We present here the efficacy of an in vitro cytotoxicity assay which can measure rapidly both apoptotic dead cells and cell growth rate, quantitatively. Using a multi-well plate reader, the fluorescence intensity of propidium iodide (PI) corresponding to dead cells and to total cells after digitonin treatment were measured in cultured human pancreatic cancer cells following exposure to etoposide. The percentage of dead cells measured by this assay was well correlated to that determined by Trypan blue staining. Furthermore, the cell growth rate determined simultaneously was also correlated to the cell number counted directly using a microscope. We demonstrate that this method, which was originally established for evaluating necrosis, could be applied to measure apoptotic cell death. Taken together, this simple assay is useful for testing the efficacy of anti-cancer agents and for investigating the molecular mechanisms of apoptosis in the cultured cells. PMID- 10463767 TI - The nitric oxide-producing activities of Scutellaria baicalensis. AB - Scutellaria baicalensis (SB) has antibacterial and antiviral activities. Nitric oxide (NO) as a potent macrophage-derived effector molecule against a variety of bacteria, viruses and tumors has received increasing attention. The objective of this study was to determine the effect of SB on the production of NO. Stimulation of mouse peritoneal macrophages with SB after the treatment of recombinant interferon-gamma (rIFN-gamma) resulted in the increased NO production. SB had no effect on NO production by itself. When SB was used in combination with rIFN gamma, there was a marked cooperative induction of NO production in a dose dependent manner. The optimal effect of SB on NO production was shown 6 h after treatment with rIFN-gamma. NO production by SB was inhibited by N(G)-monomethyl-L arginine. The increased production of NO from rIFN-(gamma) plus SB-stimulated cells was decreased by the treatment of protein kinase C inhibitor such as staurosporin. In addition, synergy between rIFN-gamma and SB was mainly dependent on SB-induced tumor necrosis factor-alpha (TNF-alpha) secretion. All the preparations of SB were endotoxin free. These results suggest that the capacity of SB to increase NO production from rIFN-gamma-primed mouse peritoneal macrophages is the result of SB-induced TNF-gamma secretion. PMID- 10463769 TI - Radiopotentiation of human brain tumor cells by sodium phenylacetate. AB - Phenylacetate (PA) inhibits the growth of tumor cells in vitro and in vivo and shows promise as a relatively nontoxic agent for cancer treatment. A recent report shows that prolonged exposure of cells to low concentrations of PA can enhance the radiation response of brain tumor cells in vitro, opening up the possibility of using this drug to improve the radiation therapy of brain tumor patients. We investigated the cytotoxicity produced by sodium phenylacetate (NaPA) alone and in combination with X-rays in SF-767 human glioblastoma cells and in two medulloblastoma cell lines, Masden and Daoy. Exposure of all three cell lines to relatively low concentrations of NaPA for up to 5 days did not enhance the subsequent cell killing produced by X-irradiation. However, enhanced cell killing was achieved by exposing either oxic or hypoxic cells to relatively high drug concentrations ( > 50-70 mM) for 1 h immediately before X-irradiation. Because central nervous system toxicity can occur in humans at serum concentrations of approximately 6 mM PA, translation of these results into clinical trials will likely require local drug-delivery strategies to achieve drug concentrations that can enhance the radiation response. The safety of such an approach with this drug has not been demonstrated. PMID- 10463770 TI - Bcl-2 plays a key role instead of mdr1 in the resistance to hexadecylphosphocholine in human epidermoid tumor cell line KB. AB - We induced tolerance to hexadecylphosphocholine (HePC) in the human epidermoid tumor cell line, KB. After 70 weeks of adaptation, the IC50 of HePC in the resistant cells KBr was 32-fold higher than in parental KB cells, and they were 30-fold more resistant to another ether lipid analogue, ET-18-OCH3. The KBr cells also showed cross-resistance to vincristine and colchicine while remaining sensitive to other chemotherapy agents. RT-PCR assays showed that expression of the multidrug resistance gene (MDR1) was positive in KBr cells, whereas the expression of GST-pi (glutathione S-transferase pi) and MRP (multidrug resistance protein) was undetectable in KBr cells. Both an immunocytochemistry test and Western blot analysis indicated that the expression of bcl-2 in KBr cells was strongly positive, while it was only mildly expressed in KB cells. Verapamil could not reverse the resistance of KBr to HePC although it is a well-known reversing agent against MDR1. Our results suggest that bcl-2 instead of MDR1 plays a major role in the resistance of KBr cells. PMID- 10463771 TI - Effects of capsaicin on induction of c-jun proto-oncogene expression in Fisher 344 rats by N-methyl-N'-nitro-N-nitrosoguanidine. AB - N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG) is a potent inducer of cellular stress leading to chromosomal aberrations, point mutations, and cell death. To study the effect of capsaicin on c-jun expression when given with MNNG to rats, Fisher-344 rats that had been administered MNNG were treated with capsaicin in their diet and organs were removed for measuring c-jun transcripts. We show that pre- or post-treatment of capsaicin relative to MNNG administration up- or down regulates (depending on the organ) c-jun expression in a consistent pattern in most organs. In fact, we found in this study that capsaicin inhibits c-jun induction, stimulated by MNNG, in the spleen, heart, stomach and lung. Since MNNG, a methylating agent, is a powerful carcinogen that is very effective in the induction of c-jun mRNA, the results suggest that capsaicin uptake in the diet could play a role in inhibition of tumorigenesis induced by MNNG. PMID- 10463772 TI - Tumor-promoting activity of 2,6-dimethylaniline in a two-stage nasal carcinogenesis model in N-bis(2-hydroxypropyl)nitrosamine-treated rats. AB - The potential promotion activity on nasal carcinogenesis of 2,6-dimethylaniline (DMA), an alpha2-adrenergic agonist metabolite of xylazine which is used for food producing animals as a sedative agent, was examined. Male F344 rats received diet containing 0 or 3000 ppm DMA for 52 weeks after initiation with N-bis(2 hydroxypropyl)nitrosamine (DHPN). Histopathological assessment showed the incidence of carcinomas in the DHPN+DMA group (33%) to be significantly elevated as compared with that for the DHPN-alone group (5%). Incidences and/or multiplicity of epithelial hyperplasias and dysplastic foci were also increased in the DHPN+DMA group. These lesions were exclusively observed in the olfactory mucosa. The lowest plasma levels of DMA in tumor- and dysplastic foci-bearing rats were 0.05 and 0.20 microg/ml, respectively. These results indicate that DHPN acts as an appropriate initiator for nasal carcinogenesis and that DMA exerts a tumor-promoting effect on the olfactory mucosa in the rat nasal cavity. PMID- 10463773 TI - The induction of smooth muscle alpha actin in a transformed rat cell line suppresses malignant properties in vitro and in vivo. AB - A normal rat fibroblast 3Y1 cell line expresses smooth muscle a actin and the expression of alpha actin is suppressed in the transformant (SR-3Y1-2) induced by a Raus sarcoma virus. Gene transfer with smooth muscle alpha actin into the SR 3Y1-2 cell line reduced growth and invasiveness in vitro, as well as tumor growth and experimental lung metastasis depending on the expression of the alpha actin. These results indicated that smooth muscle alpha actin is involved in the regulation of cell growth as well as cell motility and thus leads to the suppression of malignant phenotypes in transformed cells. PMID- 10463775 TI - The anti-proliferative effects of tyrosine kinase inhibitors towards tamoxifen sensitive and tamoxifen-resistant human breast cancer cell lines in relation to the expression of epidermal growth factor receptors (EGF-R) and the inhibition of EGF-R tyrosine kinase. AB - We have investigated the anti-proliferative effect of the tyrosine kinase inhibitors genistein, lavendustin A and 2,5-dihydromethylcinnimate (2,5-MeC) (a stable erbstatin analogue) against the epidermoid A431 cell line, the ZR-75-1 human breast cancer cell line and tamoxifen-resistant (ZR-75-9a1) and oestrogen independent (ZR-PR-LT) variants. Increased EGF-R expression by ZR-75-9a1 was associated with decreased sensitivity to genistein and 2,5-MeC whilst decreased EGF-R expression (ZR-PR-LT) cells was associated with increased sensitivity to lavendustin A and 2,5-MeC. Anti-proliferative IC50 concentrations of 2,5-MeC, but not genistein or lavendustin A, inhibited EGFR-R associated tyrosine kinase activity of ZR-75-1 cells and variants by 20-50%. PMID- 10463774 TI - Interleukin-1 receptor antagonist mRNA expression and the progression of gastric carcinoma. AB - Interleukin-1 receptor antagonist (IL-1ra), an endogeneous inhibitor of IL-1, plays an immunosuppressive role in vivo by blocking the proinflammatory effects of IL-1. In the present study, we examined whether IL-1ra expression in human gastric carcinoma correlates with tumor progression and/or metastatic potential. The reverse transcription-polymerase chain reaction was used to compare the expression of the secreted form of IL-1ra (sIL-1ra) and the intracellular form of IL-1ra (icIL-1ra) mRNA in tumor and corresponding benign tissue obtained from 38 patients with gastric carcinoma. The incidence of sIL-1ra mRNA expression was significantly higher in tumor (52%) than in corresponding benign tissue (18%) (P = 0.002). On the contrary, icIL-1ra mRNA was detected in all tumors and benign tissues. The expression of sIL-1ra mRNA by malignant tissue correlated positively with both lymph node metastasis (P = 0.008) and liver metastasis (P = 0.015). There was no association between tumor sIL-lra mRNA expression and other clinicopathologic factors. The degree of regional lymph node reaction, such as sinus histiocytosis, in tumors expressing sI-1ra mRNA was significantly weaker than that in tumors without sIL-1ra mRNA expression (5/20 vs. 12/18, P = 0.010). These results demonstrate that the altered expression of sIL-1ra by malignant tissue may be related to the progression of gastric carcinoma via modulating host immune response. PMID- 10463776 TI - Is radiosensitive cell line cross-sensitive to heat?: Effect of heat on two rat yolk sac tumor cell lines with different radiosensitivity. AB - The differences between two rat yolk sac tumor cell lines, which are of the same origin but differ in their response to irradiation, in thermal sensitivity and development of thermotolerance were investigated. A radiosensitive cell line NMT 1 is consistently less heat sensitive than the radioresistant cell line NMT-1R. The thermotolerances in NMT-1 and in NMT-1R preheated at 43 degrees C for 30 min, respectively, reached peaks at 6 h and at 12 h, but did not completely disappear at 48 h. Thermotolerance with a good correlation between both cell lines with HSP70 accumulation was observed in a different time course after a primary heating. PMID- 10463777 TI - Induction of antitumor immunity using intercellular adhesion molecule 1 (ICAM-1) transfection in mouse glioma cells. AB - We investigated the role of intercellular adhesion molecule 1 (ICAM-1) expression in tumorigenicity of gliomas and the antitumor effects of glioma cells genetically engineered to express ICAM- 1. Mouse glioma cells transfected with ICAM-1 grew in T-cell deficient mice but not in syngeneic mice. Vaccination with ICAM-1-transfected tumor cells markedly inhibited the growth of subcutaneously inoculated gliomas but not gliomas located in the brain. In vivo antibody ablation study revealed that CD4+ T, CD8+ T and NK cells were all required to produce the antitumor effect of SR/ICAM- 1. In this study, we demonstrated the therapeutic potential of vaccination with ICAM-1-overexpressing tumor cells for the control of the tumor growth. PMID- 10463779 TI - Chemical synthesis of yeast mitochondrial ATP synthase membranous subunit 8. AB - Chemical synthesis of highly hydrophobic peptides and proteins remains a challenging problem. Strong interchain associations within the peptide-resin matrix have to be overcome. A synthetic strategy for solid phase peptide synthesis is proposed, mainly based on prolonged coupling time using aprotic polar solvent mixtures. A tailored chromatographic purification was required to obtain a sample sufficiently pure for structural analysis. In this work, the total chemical synthesis of the membrane-embedded yeast mitochondrial ATP synthase subunit 8 is described. The quality of the synthetic protein was checked by electrospray mass spectrometry, its tendency to adopt alpha-helical secondary structure is evidenced by circular dichroism spectroscopy. PMID- 10463778 TI - Telomerase activity in malignant ovarian tumors with deregulation of cell cycle regulatory proteins. AB - Using a semiquantitative telomeric repeat amplification protocol assay, telomerase-positive frequencies and enzyme levels were measured. Out of 95% of 49 human ovarian tumors, the highest level of telomerase activity was observed in malignant tumors. Furthermore, by immunohistochemical staining of cell cycle regulatory proteins (pRB, p16, cyclin D1, cyclin E and p53) at the G1 checkpoint, we evaluated the relation between each protein alterations and the levels of telomerase activity. We could not demonstrate a clear relation with each molecule except for cyclin E, but suggesting that aberrant accumulation of these proteins was considered as a reason for telomerase deregulation, which may play an essential role in the pathway of telomerase regulation. PMID- 10463780 TI - Study of the conformational profile of selected unnatural amino acid residues derived from L-phenylalanine. AB - The present work reports the results of a conformational study performed on seven unnatural amino acid residues and on its natural precursor, investigated by means of computational methods at the molecular mechanics level. Amino acid residues selected for the present study are derivatives of L-phenylalanine substituted at the alpha and/or beta carbons. This series is composed of different linear analogs, including alpha-methyl, beta-methyl and beta-phenyl substituted with different stereochemistry. Analysis of the Ramachandran maps of the corresponding dipeptides in vacuo reveals their conformational preferences, to be used as guidance for the synthesis of constrained peptide analogs with desired conformational propensities. The available conformational space for every dipeptide is also analysed. PMID- 10463781 TI - Design and synthesis of a peptide derived from positions 195-244 of human cdc25C phosphatase. AB - We have designed, synthesized and purified a 51 amino acid peptide derived from an essential domain of human cdc25C phosphatase. In vivo, differential phosphorylation of this domain regulates either the induction of mitotic processes, or the checkpoint arrest of eukaryotic cells in response to DNA damage. Peptide synthesis was achieved using the stepwise Fmoc strategy and resulted in an important yield of highly pure peptide. The final peptide was identified by amino acid analysis, electrospray mass spectrometry and nuclear magnetic resonance, which revealed that one of the two methionines within the peptide was oxidized into its sulphoxide derivative We investigated whether this 51 amino acid peptide folded into secondary structures in solution by circular dichroism and observed the formation of alpha helices in TFE. Finally, we verified that this peptide could bind to its biologically relevant 14-3-3 partner in vitro by fluorescence spectroscopy. PMID- 10463782 TI - Synthesis of cyclic Herpes simplex virus peptides containing 281-284 epitope of glycoprotein D-1 in endo- or exo-position. AB - We have prepared two types of cyclopeptides containing the 281DPVG284 sequence from the 276-284 region of glycoprotein gD-1 of the Herpes simplex virus (HSV). The syntheses were performed by solid phase methodology using MBHA or BHA resin and orthogonal protection schemes. Head-to-side-chain cyclization included the N terminal part of the epitope, while side-chain-to-side-chain lactam bridge formation resulted in a peptide containing a C-terminal cycle. Peptides elongated by Cys at the N-terminal of the sequence were also prepared. Boc chemistry using Fmoc and OFm orthogonal protection was applied for on-resin cyclization. Based on the orthogonality of Bzl and cHex esters under a 1 M TMSOTf-thioanisole/TFA cleavage condition, a new approach for the cyclization on BHA-resin has also been developed. Preliminary studies on solution conformation of the cyclic peptides by CD spectroscopy indicated the importance of the location and the size of the cycle within the epitope sequence. PMID- 10463783 TI - Behaviour of dehydroalanine derivatives under hydrazinolysis conditions. Possible relevance to glycoprotein hydrazinolysis. AB - Reactions that are relevant to the cleavage, by hydrazinolysis, of O-linked oligosaccharides from glycoproteins were studied using dehydroalanine derivatives as models of the intermediates formed from O-glycosylated serine residues. Conjugate addition of hydrazine followed by cyclisation to form pyrazolidinones, if occurring during glycoprotein hydrazinolysis, could reduce the yield of released oligosaccharide. However, N-acetyldehydroalanine amide derivatives, which modelled the dehydroalanine derivatives believed to be intermediates in the hydrazinolysis of glycoproteins containing O-linked oligosaccharides, underwent conjugate addition but no cyclisation. PMID- 10463784 TI - Diurnal rhythms of fetal and maternal heart rate in the baboon. AB - To investigate the organization of diurnal rhythmicity during gestation, the relationship between daily cycles of maternal and fetal heart rate were measured in long-term studies of healthy chronically instrumented pregnant baboons. In each of six pregnancies, hourly mean values over a 168 h time series were obtained during a 7 to 10 day interval between 135 and 160 days of gestation. Data were modeled by a least squares fit to a cosine function with a period of 24 h. Hourly mean heart rate in the fetus ranged from 161 to 172 bpm (167.9+/-0.6 bpm), and the mother from 105 to 125 bpm (107.9+/-1.4 bpm). The amplitude of the daily fluctuations were 15 to 25 bpm for the fetuses and 25 to 60 bpm for the mothers. The relation between time series data and model estimates were significant (P < 0.001) in all cases with aggregate r2 = 0.747 for fetuses and 0.737 for the mothers. On average the time of day of the peak in fetal heart rate (15:05+/-0.3 h) was about 45 min after the maternal peak (14:21+/-0.4 h). This phase delay was significant (t = 2.63, P < 0.05). There was significant (P < 0.01) diurnal periodicity for each of six parameters used to assess different aspects of fetal heart rate variability with peak variability at night (23:00 to 2:00). Thus, during the latter third of pregnancy in both the maternal and fetal baboon 24 h periodicities of heart rate are present with peak rates in the midafternoon. The daily rhythms in fetal heart rate are linked with periodicities in maternal heart rate with a phase delay in the majority of cases. The synchrony of 24 h fluctuations in rate with parameters of rate variability is consistent with diurnal input into the fetal autonomic nervous system. PMID- 10463785 TI - Oral glucose and venepuncture reduce blood sampling pain in newborns. AB - The objectives of this study were to measure pain symptoms in healthy fullterm newborns undergoing routine blood sampling with different methods. The 120 study subjects were randomly allocated to one of four groups with 30 babies in each, namely venepuncture or heel stick, with or without oral glucose administration. Pain was assessed from the duration of crying within the first 3 min, the Premature Infant Pain Profile (PIPP) and changes in heart rate. When the babies received 1 ml 30% glucose prior to skin puncture there was no significant difference between the heel stick and venepuncture group either in mean crying time (12.9 and 11.6 s, respectively) or in PIPP score (3.9 and 3.3). When no glucose was given crying time was 57.3 s in the heel stick group and 26.8 s in the venepuncture group (P = 0.0041) and the mean PIPP scores were 8.4 and 6.0, respectively (P = 0.0458). This study suggests that if oral glucose is given prior to skin puncture the choice of blood sampling method has no impact on the pain symptoms. PMID- 10463786 TI - Spontaneous awakenings from sleep in the first year of life. AB - Spontaneous awakenings from nocturnal sleep were studied in a sample of 48 healthy infants (M = 26, F = 22), in four age groups (1 to 7 weeks, 8 to 15 weeks, 17 to 22 weeks, 25 to 54 weeks). Consistent with previous data, the number of awakenings is reported less frequently at later ages, owing to a lower frequency of awakenings out of REM sleep. Like young adults, infants in all age groups awake more often from REM than from quiet sleep (QS); this is particularly evident in the first 6 months of life, less so in the second. The duration of the bouts of wakefulness following awakenings remains stable with age. Awakenings out of QS are followed by longer periods of wakefulness than those out of REM sleep, although in older infants the duration is considerably reduced. Night sleep first shows a decrease in the number of awakenings out of REM sleep and then continues after the sixth month of life with the shortening of the wakefulness after awakenings out of QS. In the two younger groups, the distribution of the awakenings shows two main peaks and one main peak differently located during the night; a polymodal pattern appears in group 3, and is even more evident in group 4. It should be stressed that several changes as a function of age occur in the second 6 months of life, indicating this as a period of intense developmental change in sleep-wake rhythms. PMID- 10463787 TI - Cerebral blood oxygenation changes induced by auditory stimulation in newborn infants measured by near infrared spectroscopy. AB - Recent neuronal activation studies on newborns using functional MRI or near infrared spectroscopy (NIRS) have suggested that the increase in O2 consumption accompanying neuronal activation exceeds the increase in O2 delivery in the visual cortex during photic stimulation. In the present study, we evaluated the cerebral blood oxygenation (CBO) changes induced by auditory stimulation in the frontal lobe of newborns using NIRS. We studied 28 newborns; the postnatal age at CBO measurements was 3.1+/-0.3 days (mean+/-S.E.M.). We measured concentration changes of deoxyhemoglobin (Deoxy-Hb), oxyhemoglobin (Oxy-Hb), and total hemoglobin (Total-Hb) induced by auditory (music) stimulation in the bilateral frontal lobes of the newborns. Twenty-six (92.9%) out of 28 subjects showed increases of Oxy-Hb and Total-Hb during the stimulation. In these subjects, 17 (60.7%) subjects showed an increase of Deoxy-Hb associated with increases of Oxy Hb and Total-Hb, while nine (32.1%) subjects showed a decrease of Deoxy-Hb. Although the direction of the Deoxy-Hb differed, these two groups did not differ for Oxy-Hb and Total-Hb (P > 0.05). Two (7.1%) subjects showed other changes. The frontal lobe of newborns shows CBO responses similar to those observed in the visual cortex, specifically neuronal activation causes an increase of Deoxy-Hb associated with increases of Oxy-Hb and Total-Hb. These results support the hypothesis that increments in O2 consumption exceed increments in O2 delivery during neuronal activity in newborns. PMID- 10463788 TI - Electrocortical brain activity during hypoxia and hypotension in anesthetized newborn lambs. AB - Blood gas and blood pressure disturbances do influence cerebral blood flow in newborns. To what extent cerebral blood flow changes affect electrocortical brain activity remains uncertain. We studied the effect of severe hypoxia and hemorrhagic hypotension on carotid artery blood flow and electrocortical brain activity in newborn anesthetized lambs. During hypoxia carotid artery blood flow increased significantly, whereas electrocortical brain activity remained unchanged. The hemorrhagic hypotension study showed that the lower limit of the autoregulatory ability of the cerebral vascular bed was 60 mmHg. Electrocortical brain activity however remained stable until mean aortic pressure had dropped below 30 mmHg, carotid artery blood flow below 10.6 ml/kg/min, and cerebral oxygen delivery below 1.4 ml/kg/min. PMID- 10463789 TI - The development of preterm infants' breastfeeding behavior. AB - Extensive scientific data are available on the development of sucking in term infants, but corresponding knowledge about preterm infants has been lacking. The aim was to describe the development of preterm infants' behavior at the breast, according to postmenstrual and postnatal age. Mothers co-operated as data collectors using the Preterm Infant Breastfeeding Behavior Scale for observations. Seventy-one singletons (born after 26.7-35.9 gestational weeks) were studied prospectively. Mothers received regular advice about breastfeeding and guidance about interaction and appropriate stimulation. Breastfeeding was initiated at a postmenstrual age (PMA) of 27.9-35.9 weeks. Mothers provided 4321 records of infants' behavior, 70% of the total breastfeeding sessions during the infants' hospital stay. Irrespective of PMA, the infants responded by rooting and sucking on the first contact with the breast. Efficient rooting, areolar grasp and latching on were observed at 28 weeks, and repeated bursts of > or = 10 sucks and maximum bursts of > or = 30 sucks at 32 weeks. Nutritive sucking appeared from 30.6 weeks. Sixty-seven infants were breastfed at discharge. Fifty-seven of them established full breastfeeding at a mean PMA of 36.0 weeks (33.4-40.0 weeks). Their early sucking behavior is interpreted as the result of learning, enhanced by contingent stimuli. We therefore suggest that guidelines for initiation of breastfeeding in preterm infants should be based on cardiorespiratory stability, irrespective of current maturity, age or weight. PMID- 10463790 TI - Cardiorespiratory responses to bidirectional tilts in infants. AB - Prior research in newborns has shown that head-up and head-down tilting elicits sustained increases and decreases in heart rate, respectively. Other studies in older infants have suggested that the pattern of heart rate responses to head-up tilting varies with risk for sudden infant death syndrome (SIDS). In this study, heart and respiratory rate changes following bidirectional tilting were recorded in sleeping infants on Day 1 or 2 of life, and during the period of maximum risk for SIDS, at 2 and 4 months of age. Newborns show increases in heart rate following 30 degrees head-up tilts and decreases in heart rate to 300 head-down tilting. Respiratory rates decreased to head-up tilting but did not change significantly to head-down tilting. While respiratory rate changes at 2 and 4 months of age are comparable to those of newborns, and decreases in heart rate to head-down tilting are similar across ages, sustained elevations in heart rate following head-up tilting are no longer apparent at the older ages. These results are consistent with the hypothesis that, during the period of maximum risk for SIDS, infants may have reduced ability to compensate for challenges that lead to decreases in blood pressure. PMID- 10463791 TI - Plasticity in the tectum of Xenopus laevis: binocular maps. AB - Xenopus frogs exhibit dramatic changes in the binocular projections to the tectum during a critical period of development. Their eyes change position in the head, moving from lateral to dorsal and creating an increasing region of binocular overlap. There is a corresponding shift of binocular projections to the tectum that keeps the two eyes' maps in register with each other throughout this period. The ipsilateral input is relayed via the nucleus isthmi. Two factors bring the ipsilateral projection into register with the contralateral projection. First, chemoaffinity cues establish a crude topographic map beginning when the shift of eye position begins. Approximately 1 month later, visual cues bring the ipsilateral map into register with the contralateral map. The role of visual input is demonstrated by the ability of the axons that bring the ipsilateral eye's map to the tectum to reorganize in response to a surgical rotation of one eye and to come into register with the contralateral eye's map. This plasticity can be blocked by NMDA receptor antagonists during the critical period. In normal adults, reorganization is minimal. Eye rotation fails to induce reorganization of the ipsilateral map. However, plasticity persists indefinitely in animals that are reared in the dark, and plasticity can be restored in normally-reared animals by treatment with NMDA. The working model to explain this plasticity posits that correlated input from the two eyes triggers opening of NMDA receptor channels and initiates events that stabilize appropriately-located isthmotectal connections. Specific tests of this model are discussed. PMID- 10463792 TI - The neurobiology of startle. AB - Startle is a fast response to sudden, intense stimuli and probably protects the organism from injury by a predator or by a blow. The acoustic startle response (ASR) of mammals is mediated by a relatively simple neuronal circuit located in the lower brainstem. Neurons of the caudal pontine reticular nucleus (PnC) are key elements of this primary ASR pathway. The ASR in humans and animals has a non zero baseline, that is, the response magnitude can be increased or decreased by a variety of pathological conditions and experimental manipulations. Therefore, the ASR has been used as a behavioral tool to assess the neuronal basis of behavioral plasticity and to model neuropathological dysfunctions of sensorimotor information processing. Cross-species examples for the increase of the ASR magnitude are sensitization, fear-potentiation and drug-induced enhancement. Examples for the reduction of the ASR magnitude are habituation, prepulse inhibition, drug-induced inhibition and the attenuation by positive affect. This review describes the neuronal basis underlying the mediation of the ASR, as well as the neuronal and neurochemical substrates of different phenomena of enhancement and attenuation of the ASR. It also attempts to elucidate the biological background of these forms of behavioral plasticity. Special emphasis is put on the potential relevance of ASR modulations for the understanding of human psychiatric and neurological diseases. PMID- 10463793 TI - The synaptic pharmacology underlying sensory processing in the superior colliculus. AB - The superior colliculus (SC) is one of the most ancient regions of the vertebrate central sensory system. In this hub afferents from several sensory pathways converge, and an extensive range of neural circuits enable primary sensory processing, multi-sensory integration and the generation of motor commands for orientation behaviours. The SC has a laminar structure and is usually considered in two parts; the superficial visual layers and the deep multi-modal/motor layers. Neurones in the superficial layers integrate visual information from the retina, cortex and other sources, while the deep layers draw together data from many cortical and sub-cortical sensory areas, including the superficial layers, to generate motor commands. Functional studies in anaesthetized subjects and in slice preparations have used pharmacological tools to probe some of the SC's interacting circuits. The studies reviewed here reveal important roles for ionotropic glutamate receptors in the mediation of sensory inputs to the SC and in transmission between the superficial and deep layers. N-methyl-D-aspartate receptors appear to have special responsibility for the temporal matching of retinal and cortical activity in the superficial layers and for the integration of multiple sensory data-streams in the deep layers. Sensory responses are shaped by intrinsic inhibitory mechanisms mediated by GABA(A) and GABA(B) receptors and influenced by nicotinic acetylcholine receptors. These sensory and motor-command activities of SC neurones are modulated by levels of arousal through extrinsic connections containing GABA, serotonin and other transmitters. It is possible to naturally stimulate many of the SC's sensory and non-sensory inputs either independently or simultaneously and this brain area is an ideal location in which to study: (a) interactions between inputs from the same sensory system; (b) the integration of inputs from several sensory systems; and (c) the influence of non sensory systems on sensory processing. PMID- 10463794 TI - The dopaminergic innervation of the avian telencephalon. AB - The present review provides an overview of the distribution of dopaminergic fibers and dopaminoceptive elements within the avian telencephalon, the possible interactions of dopamine (DA) with other biochemically identified systems as revealed by immunocytochemistry, and the involvement of DA in behavioral processes in birds. Primary sensory structures are largely devoid of dopaminergic fibers, DA receptors and the D1-related phosphoprotein DARPP-32, while all these dopaminergic markers gradually increase in density from the secondary sensory to the multimodal association and the limbic and motor output areas. Structures of the avian basal ganglia are most densely innervated but, in contrast to mammals, show a higher D2 than D1 receptor density. In most of the remaining telencephalon D1 receptors clearly outnumber D2 receptors. Dopaminergic fibers in the avian telencephalon often show a peculiar arrangement where fibers coil around the somata and proximal dendrites of neurons like baskets, probably providing them with a massive dopaminergic input. Basket-like innervation of DARPP-32-positive neurons seems to be most prominent in the multimodal association areas. Taken together, these anatomical findings indicate a specific role of DA in higher order learning and sensory-motor processes, while primary sensory processes are less affected. This conclusion is supported by behavioral findings which show that in birds, as in mammals, DA is specifically involved in sensory-motor integration, attention and arousal, learning and working memory. Thus, despite considerable differences in the anatomical organization of the avian and mammalian forebrain, the organization of the dopaminergic system and its behavioral functions are very similar in birds and mammals. PMID- 10463795 TI - A comparison of amino acid distance measures using procrustes analysis. AB - Four methods to convert an amino acid similarity matrix into a metric distance matrix were compared using procrustes analysis. Procrustes analysis is a rotational fit technique, which assesses the degree of fit between two configurations after optimal matching of the two matrices under translation, rotation and scaling. The analysis shows that not all conversion methods are equivalent, but that the results of two methods are more similar than the others. PMID- 10463796 TI - An algorithm for finding the linear region in a nonlinear data set. AB - Finding the linear reportable range is an important procedure for each method in clinical chemistry. One is often called upon to limit the reportable range in order to find the linear region. Limiting the reportable range by visual techniques is subjective, may introduce bias and is not programmable. Using Kroll and Emancipator's polynomial method for linearity, we compare the residuals of a test to determine whether eliminating a point from one end or the other of the data set worsens or improves the data sets' linearity. In an example of urinary cortisol, the root mean squares of the residuals improve by 2% when the lowest point is removed, 39% when the highest point is removed and 82% when the two highest points are removed. The latter data set is the most linear. PMID- 10463797 TI - Interactive dynamic navigation for virtual endoscopy. AB - Complex anatomical information can be obtained from a 3D radiologic image by navigating through it in a manner similar to an endoscopic examination. Real-time computation of 'virtual' endoscopic views, however, is needed to permit interactive navigation. We present a fast volume-rendering method for computing such views. Our method, motivated by the temporal-coherence concept, performs dynamic volume rendering at interactive frame rates. Results demonstrate the method's efficiency and accuracy. Also, our method constitutes part of a complete virtual-endoscopic system we have devised. This system is illustrated for 3D pulmonary analysis. PMID- 10463798 TI - Automated registration of multimodal brain image sets using computer vision methods. AB - We present a new method of registering three dimensional image volumes of the brain of a given patient acquired at different times or with different imaging modalities, or both. Registration is an essential requirement for fusing the data from the two image sets so as to either increase the available information by exploiting complementary imaging modalities, or to measure small changes over time for prognostication, disease assessment, etc. The new technique exploits an external, removable, remountable reference frame which is attached to the head. Computer vision techniques are used to determine the positions of fiducial marks in every image. The transformation required to map each image of one image volume onto the other image volume is developed using the theory of quaternions. The results indicate that the new technique is robust and practical in a clinical setting. PMID- 10463799 TI - Long-term treatment careers and outcomes of previously untreated alcoholics. AB - OBJECTIVE: The aim of this project was to describe treatment selection and outcomes over an 8-year follow-up period for 466 individuals who had drinking problems and had not yet received formal treatment at baseline. METHOD: By the 8 year follow-up, individuals had self-selected into one of four groups: no treatment (n = 78); completed treatment (help was received only in Years 1-3 of follow-up; n = 230); additional treatment (help was received in Years 1-3, with more help in Years 4-8; n = 134); and delayed treatment (no help was received until Years 4-8 of follow-up; n = 24). RESULTS: Compared with individuals who remained untreated, the completed treatment group had more severe drinking problems and depression at baseline, but better drinking outcomes at both the 3- and 8-year follow-ups. At 3 years, the additional treatment group perceived their drinking problem as being more serious than did the untreated group and was more likely to have drinking-related problems; by 8 years, individuals who had obtained additional treatment were more likely to be abstinent, but still perceived their drinking problems as being more serious. Compared to individuals who completed treatment in Years 1-3, additional treatment group members had more severe drinking and functioning problems at 3 and 8 years. More involvement with formal outpatient treatment or AA was associated with more improvement on drinking indices. CONCLUSIONS: Persons with alcohol use disorders who elect to enter formal treatment or AA relatively soon after acknowledging their drinking problems experience better drinking-related outcomes than do those who receive no help or who delay receiving help. Accordingly, referral processes should ensure that problem drinkers enter self-help or formal treatment quickly. PMID- 10463800 TI - Cross-cultural applicability in international classifications and research on alcohol dependence. AB - OBJECTIVE: Underlying cultural differences in the meaning of alcohol problems and dependence symptoms can lead people from different societies to systematically vary in their responses to a diagnostic interview, in ways that may be difficult for researchers to quantify or control. We identify four different ways that cultures can vary in their accounts of dependence symptoms, potentially leading to problems with the cross-cultural applicability of diagnostic criteria: (1) in terms of thresholds of symptom severity, or the point at which respondents from different societies recognize a symptom of dependence as something serious; (2) in the problematization of drinking-related states, or whether the symptoms described in official nomenclature on addiction even count as alcohol problems in all cultures; (3) in causal assumptions about how alcohol-related problems arise; and (4) in the extent to which there exist culture-specific manifestations of symptoms not adequately captured by official disease nomenclature. METHOD: Comparable data on the meaning of alcohol dependence criteria were collected from key informants in nine sites worldwide under the auspices of the WHO/NIH Cross Cultural Applicability Research Project. Qualitative analysis compares and contrasts descriptions of ICD-10 dependence criteria across sites along the above four dimensions of cultural variation. RESULTS: While descriptions of dependence symptoms were quite similar among key informants from sites that share norms around drinking and drunkenness, they varied significantly in comparisons between sites with markedly different drinking cultures. Contrary to expectation, descriptions of physical dependence criteria appeared to vary across sites as much as the more subjective symptoms of psychological dependence. CONCLUSIONS: Problems with the cultural applicability of international nomenclature warrant careful consideration in future comparative research on addiction, although comparisons of dependence made across some cultural boundaries are likely to be much more problematic than comparisons made across others. Findings on dependence should be interpreted in light of what is known about the drinking cultures and norms of the societies involved. Future nosologies and diagnostic interview schedules should take into account a broad base of cultural experiences in conceptualizing alcohol dependence, in developing criteria and operationalizations and in determining the diagnostic significance of these. PMID- 10463801 TI - Resolving alcohol-related problems with and without treatment: the effects of different problem criteria. AB - OBJECTIVE: Prior research indicates that the majority of people who resolve an alcohol problem do so without treatment. Further, a substantial proportion return to moderate drinking. One of the concerns with these findings, however, is that the definition of prior alcohol problems may reflect a less severe problem criterion. Thus, it could be argued that the majority of people "recover" from alcohol problems without treatment because a substantial proportion of these individuals did not actually have a severe alcohol problem. METHOD: This article systematically replicates earlier findings regarding the predominance of alcohol recoveries without treatment, using data from the 1994 Canadian Alcohol and Drug Survey. Further, variations in estimates of the proportion of resolutions from alcohol problems both with and without treatment, and to abstinence and moderate drinking, were assessed when different definitions of prior alcohol problems were employed. RESULTS: Of 9,892 adult lifetime drinkers, 2,177 had experienced at least one problem related to their alcohol consumption and, of these, 885 (57.2% male) had experienced no problems in the last year. Estimates of the prevalence of nontreatment recoveries ranged from 87.5% to 53.7% depending on the stringency of the definition of prior alcohol problems employed. CONCLUSIONS: Although a significant proportion of individuals recover from alcohol problems without treatment, such recoveries appear less common among individuals with more severe alcohol problems. PMID- 10463802 TI - Heavy drinking and problems among wine drinkers. AB - OBJECTIVE: The purpose of this study was to see whether adults who drank wine exclusively would have lower quantity-frequency scores, less intoxication and fewer alcohol-related problems than those who drank other types of alcohol. METHOD: Data were analyzed from a national survey of the use of alcohol and other drugs conducted in Canada in 1994 (N = 8,758). RESULTS: Wine only drinkers were less often intoxicated than other drinkers. They also had lower quantity frequency scores and reported fewer alcohol-related problems than other drinkers. These results were maintained when age, gender and quantity-frequency were taken into account. CONCLUSIONS: Wine only drinkers drink less than other types of drinkers and have fewer problems. As wine is often drunk with meals there is probably more social control over drinking. Also, since wine drinkers drink with meals their blood alcohol concentrations may be lower. PMID- 10463803 TI - Alcoholics with and without social phobia: a comparison of substance use and psychiatric variables. AB - OBJECTIVE: While alcoholics with social phobia comprise a substantial portion of the alcoholic population, little is known about how they differ from alcoholics without social phobia in their substance use and psychiatric health. The present study was conducted to examine baseline differences between alcoholics with and without social phobia on substance use and psychiatric variables. METHOD: Alcoholics without social phobia (n = 397) were chosen to match those with social phobia (n = 397) on several variables, including age and gender. All subjects were participants in Project MATCH, a large clinical client-treatment matching study. RESULTS: Exploratory/Confirmatory analyses revealed that alcoholics with social phobia had higher scores on the alcohol dependence scale and endorsed more dependence symptoms on the SCID, although they did not drink greater amounts or more often than alcoholics without social phobia. They also reported drinking in order to improve sociability and enhance functioning more than did the comparison group. Alcoholics with social phobia were more likely to conform to social norms than alcoholics without social phobia. They also had more symptoms of depression as indicated by higher scores on the Beck Depression Inventory and higher incidence of a major depressive episode from the C-DIS. CONCLUSIONS: Alcoholics with social phobia enter treatment with some problems that are more severe than those expressed by alcoholics without social phobia. Whether these problems affect treatment efficacy is an important area for future research. PMID- 10463804 TI - Transition into adolescent problem drinking: the role of psychosocial risk and protective factors. AB - OBJECTIVE: To establish the role of psychosocial risk and protective factors in cross-sectional variation in adolescent problem drinking, and in the transition into problem drinking over time. METHOD: The data were from a four-wave (1989 1992) longitudinal study of 1,591 adolescents in a large, urban school district. School district officials selected schools for the study with an aim toward maximizing representation of minority students from inner-city areas. At Wave 1, all students in Grades 7, 8, and 9 were asked to participate. RESULTS: Both psychosocial risk factors (such as low expectations for success, peer models for substance use, and poor school performance) and psychosocial protective factors (such as intolerance of deviance, peer models for conventional behavior, and involvement in prosocial activities) account for significant cross-sectional variation in adolescents' involvement in problem drinking, as indicated by more frequent drunkenness and more numerous instances of alcohol-related problems. They also account for significant variation in the timing of transition into problem drinking during adolescence. Higher risk and lower protection are each associated with greater problem use of alcohol. Among adolescents who are not problem drinkers, higher risk and lower protection accelerate the likelihood of becoming a problem drinker in subsequent years. Protection also moderates the impact of risk in the cross-sectional account of involvement in problem drinking, but protective factors appear not to play a moderating role in the longitudinal account of the transition into problem drinking. Findings were similar for males and females and among white, black and Hispanic adolescents. CONCLUSIONS: Protective factors play an independent role in accounting for adolescent involvement in problem drinking and in the transition into problem drinking in adolescence. Intervention efforts to enhance protection, especially for adolescents who are exposed to risk, should supplement efforts to reduce risk. PMID- 10463805 TI - Social and coping reasons for drinking: predicting alcohol misuse in adolescents. AB - OBJECTIVE: Motivational models of alcohol consumption suggest a positive relationship between reasons for drinking and the amount of alcohol consumed. The present study examined race, gender, and age as moderators of the relationship between social and coping motives and alcohol misuse in black and white adolescents. METHOD: A representative population sample (N = 699) of male and female (54%) adolescents between the ages of 13 and 16 was recruited using a random-digit-dial telephone procedure. Six face-to-face interviews with subjects and their families were carried out at approximately yearly intervals. Information gathered assessed alcohol use, social and coping motives for drinking, and psychological distress. RESULTS: Multiple analyses, including both cross-sectional and longitudinal logistic regression analyses and survival analysis were used to examine the relationship of drinking motives to adolescent alcohol misuse. Contrary to our predictions, social motive was a somewhat better predictor of alcohol misuse than was coping motive, particularly during mid- to late adolescence. However, there was some limited evidence of a significant relationship between coping motives and alcohol misuse in the mid-adolescent age group. Some support was found for racial differences such that social motives are better predictors of alcohol misuse among whites than among blacks and coping motives are better predictors among blacks. Few gender differences were found in the relationship of drinking motives and alcohol misuse. CONCLUSIONS: These findings suggest a stronger tendency for social and coping motives to influence alcohol misuse during mid to late than in early adolescence. Research examining the development of motives and the mechanisms by which they influence drinking behavior is needed. PMID- 10463806 TI - The role of neurocognitive abilities in coping with adolescent relapse to alcohol and drug use. AB - OBJECTIVE: Neurocognitive abilities are hypothesized to influence coping with temptations to relapse to alcohol and drug use. In particular, a moderator model was proposed whereby neurocognitive abilities moderate the influence of coping strategies on treatment outcome. METHOD: Participants (N = 79) were consecutive admissions to inpatient alcohol and drug treatment centers in the San Diego, California. area, (age range = 14 to 18 years; 41% female). Treatment programs were abstinence focused, with an average inpatient stay of 5 weeks. Participants met DSM-III-R lifetime criteria for alcohol abuse or dependence and abuse or dependence of at least one other substance. Follow-up assessments were conducted 1 and 2 years following discharge, including a battery of neuropsychological tests, coping questionnaire, and detailed assessment of alcohol and other drug involvement. RESULTS: The interaction between general intelligence and coping diversity significantly predicted alcohol and drug use 1 year later (F = 10.71, 1/72 df, p <.005, R2 change =.08), even when controlling for current use. Attention-coping and problem solving-coping interactions also significantly predicted outcome, but not above the intelligence-coping interaction. CONCLUSIONS: Teens with poor neurocognitive abilities used little or no alcohol and drugs if good coping skills were evidenced, but used more alcohol and drugs if their coping skills were poor. This association between coping skills and outcome did not exist for teens with better neurocognitive abilities. Clinically, teens with poorer neurocognitive skills may maximally benefit from coping skills training programs, and neuropsychological evaluations may be particularly helpful in addressing these individual needs in treatment. PMID- 10463807 TI - Neuropsychological characteristics in children of alcoholics: familial density. AB - OBJECTIVE: The purpose of high-risk studies is to find characteristics that allow the identification of subjects with a higher vulnerability to alcoholism. The aim of this research was to verify if the familial density criterion is useful for subtyping children of alcoholics with different neuropsychological characteristics. METHOD: A battery of neuropsychological tests was administered to 102 boys and girls of 7-15 years of age; 66 were children of alcoholics with a high (n = 32) and low (n = 34) familial density of alcoholism, and 36 were children of nonalcoholic fathers with a negative family history of the disorder. The battery included tests to assess attention, visuospatial abilities and frontal functions. RESULTS: MANCOVAs showed that high-density children scored lower than children of nonalcoholic fathers in attentional and visuospatial tasks. There were no differences between low-density and negative family history children in these cognitive domains. CONCLUSIONS: These results suggest that children of alcoholics are not a homogeneous group. Children with multigenerational alcoholism, but not children with an alcoholic father, showed reduced performance in specific cognitive areas. PMID- 10463808 TI - The effect of victim impact panels on DUI/DWI rearrest rates: a twelve-month follow-up. AB - OBJECTIVE: Various interventions have been implemented as components of sentencing for driving under the influence of alcohol. This study assessed the effectiveness of the Victim Impact Panel (VIP) in reducing the probability of an offender being rearrested for drinking and driving. METHOD: The study was conducted in an urban/suburban county in the Southeastern United States. Rearrest rates of DUI offenders who attended a VIP presentation as part of their sentencing (n = 404) were compared with an equivalent comparison group (n = 431) who did not have the VIP presentation. Arrest records of offenders were searched and rearrest rates (%) were calculated for the periods 0-6 months, 7-12 months, and 0-12 months after the arrest. RESULTS: Chi-square cross-tabulations indicate that rearrest rates were lower for the VIP group than the comparison group in all categories. Three categories where the differences were significant and of the greatest magnitude were white men, ages 26-35 years, and one prior DUI arrest. Additionally, logistic regression was used to compare the importance of specific independent variables on rearrest. Whether or not a subject was in the VIP group was the most powerful contributor to the results. CONCLUSIONS: After considering alternative explanations for the results, we conclude that the VIPs can be a cost effective way of reducing the probability of rearrest in DUI offenders. When costs of DUI in human misery and dollars are considered, the potential benefits of large-scale implementation of VIP programs appear to be well worth the effort. PMID- 10463809 TI - Alcohol risk reduction for fraternity and sorority members. AB - OBJECTIVE: The primary objective of this study is to evaluate the effectiveness of "Talking about Alcohol and Drugs... Among Greeks" (TAAD), a lifestyle risk reduction program that is research- and theory-based, protocol-driven, and targeted for fraternity and sorority members. METHOD: One fraternity and two sororities on five campuses participated in the program. A total of 780 participants completed pre- and posttest questionnaires. Posttest data were collected 1 academic year after pretest data collection. RESULTS: Results indicate that the program decreased positive attitudes toward alcohol consumption among program participants, with participants in the true experimental condition indicating greater disagreement than control participants (F = 3.05, 2/701 df, p < .05). Belief in myths about the etiology of alcoholism was reduced among experimental participants who did not actually attend the program, with those participants indicating greater disagreement than control or true experimental participants (F = 10.92, 2/702 df, p < .0001). The program had no apparent effect on alcohol consumption by experimental participants. CONCLUSIONS: The program's ability to influence behavior was hindered by systemic problems, such as trainer credibility and implementation infidelity, and probable participant psychological reactance. Training by professionals and mandatory attendance would set the stage for improved program effectiveness. Beyond that, an approach that emphasized how high-risk drinking norms jeopardize the goals of the fraternities and sororities might be better received than the current approach, which focuses on how individual attitudes, beliefs and behaviors lead to alcohol-related problems and alcoholism. PMID- 10463810 TI - The role of alcohol in physical assault perpetration and victimization. AB - OBJECTIVE: This study examines the influence of lifetime alcohol use and drinking in-the-event on the risk of physical assault perpetration and victimization while controlling for the effects of additional demographic and personality characteristics known to be associated with violence or alcohol use. METHOD: Secondary analyses were performed on data collected for the 1990 National Alcohol Survey, which included in-person interviews with 2,058 adults (58% female) residing within the 48 contiguous United States. RESULTS: Approximately 11.5% of the sample reported having committed a physical assault, and 16.6% of the respondents reported having been the victim of a physical assault, since the age of 12 years. Gender, age, education, lifetime drinking history and an interaction between age and impulsivity were associated with physical assault perpetration. Marital status, impulsivity, and life-time drinking history were associated with physical assault victimization. Analyses performed on the subsample of individuals reporting a physical assault indicated that drinking-in-the-event by both the perpetrator and victim was associated with men's, and not women's, experiences. CONCLUSIONS: The results support a strong and stable relationship between alcohol use and physical assault. Previous findings regarding men and alcohol-related aggression are supported, but the results contradict past conclusions pertaining to alcohol and women's victimization. PMID- 10463811 TI - Drinking and marital aggression in newlyweds: an event-based analysis of drinking and the occurrence of husband marital aggression. AB - OBJECTIVE: Research suggests that domestically violent men are more likely to be heavy drinkers than nonviolent men. However, very little data has addressed the relationship between the occurrence of a domestic violence episode and the co occurrence of husband or wife alcohol consumption. The purpose of this study was to examine the relationship between husband and wife drinking and whether the most serious conflict involved verbal aggression, moderate physical aggression or severe physical aggression. METHOD: Newlywed couples (n = 366) who were assessed with respect to risk factors for marital violence at the time of marriage were interviewed at the end of their first year of marriage. These in-person interviews focused on characteristics (location, presence of other people, husband drinking, wife drinking) of the most severe verbal conflict, and the first and the most severe episode of physical aggression. RESULTS: After controlling for individual difference risk factors including husband and wife drinking patterns, husband drinking was more likely in episodes of physical vs episodes of verbal aggression. This finding was consistent in both husband and wife, and consistent in both between-groups and within-groups analyses. Wife drinking was largely unrelated to occurrence or severity of violence. CONCLUSIONS: These results support the contention that alcohol use may contribute to the occurrence and/or severity of domestic violence, particularly in the early stages of marriage. PMID- 10463812 TI - Physical dependence after benzodiazepine treatments in rats: Comparison of short and long treatments with diazepam and lorazepam. AB - OBJECTIVE: This study examined the development of physical dependence after different durations of treatments with two benzodiazepines (diazepam and lorazepam). METHOD: Increased excitation in the central nervous system during a 2 week withdrawal period after 4-day and 4-week treatments with diazepam and lorazepam was examined with an EEG threshold method in male rats. Increased excitation was measured as a decreased sensitivity to hexobarbital (i.e., increased threshold doses). The concentrations of hexobarbital in two different brain regions, serum, fat and muscle tissue after 4-week treatment with diazepam were determined with a high-pressure liquid chromatography method. RESULTS: The duration of withdrawal was influenced by the duration of treatment but the maximum level of withdrawal excitation was similar for both drugs. Equieffective doses of diazepam (20 mg/kg) and lorazepam (2 mg/kg) induced similar patterns of withdrawal excitability after both treatments. The brain concentrations of hexobarbital were significantly higher on Days 1 and 3 of withdrawal after diazepam treatment. Significant correlations between the threshold doses and brain concentrations were found on Day 1, but these correlations disappeared on day 3. At the same time, a difference between the concentrations of hexobarbital in different brain areas emerged. CONCLUSIONS: The duration of treatment had a minor influence on the pattern of withdrawal excitation. Equieffective doses of diazepam and lorazepam induced comparable withdrawal excitability indicating no significant difference in their potential to induce physical dependence. The time dependent change in the hexobarbital concentrations in the brain suggests that withdrawal excitation after diazepam treatment is a complex phenomenon probably involving several different systems at different times. PMID- 10463813 TI - Development of dosing guidelines for reaching selected target breath alcohol concentrations. AB - OBJECTIVE: This study evaluated gender-specific ethanol dosing protocols that were designed to result in one of two peak breath alcohol concentrations (BrACs)- 0.07 or 0.10 g/2101. Inter- and intrasubject variability in BrAC were assessed and several possible methods for reducing variability in BrAC were evaluated. METHOD: Subjects (16 women, 16 men, ages 21-30 years) were studied after low (women 0.49 g/kg, men 0.53 g/kg consumed over 10 minutes) and high (women 0.81 g/kg, men 0.89 g/kg consumed over 20 minutes) ethanol doses, consumed following a 4-hour fast. All subjects were regular drinkers. RESULTS: Mean (+/-SD) peak BrACs actually achieved were 0.069+/-0.011 g/2101 after the low dose, and 0.105+/-0.014 g/2101 after the high dose. Mean values for peak BrAC, time to peak BrAC and area under the curve were not statistically significantly different between genders at either dose. BrACs varied by as much as twofold between subjects after equivalent gender and body weight adjusted doses. There was some reproducibility of ethanol pharmacokinetic parameters over dose and time in men, but not in women. CONCLUSIONS: The doses used resulted in equivalent mean ethanol exposures for women and men at each dose, with mean peak BrACs that closely approached the targets, but there was substantial inter- and intrasubject variability in ethanol pharmacokinetics. PMID- 10463814 TI - Studies of factors in relapse to alcohol, drug and nicotine use: a critical review of methodologies and findings. AB - OBJECTIVE: To critically review the methodologies used to study relapse in substance abusers and compare some of the findings these methodologies have produced. METHOD: Representative studies that have used retrospective, prospective, or near real time methodologies to study antecedents or consequences of relapse are presented and the strengths and weaknesses of each methodology are reviewed. Limitations in the ability of substance abusers to accurately report internal states are considered. Degree of convergence in findings across methodologies is examined. RESULTS: Retrospective reports can provide perhaps the most detailed accounts of circumstances and experiences leading up to relapse, but are also more likely to be inaccurate due to the limitations of memory. Prospective studies are less likely to be affected by memory problems, but may not capture factors that immediately precede relapse. The near real time methodology, which uses palm-top computers to gather data, has many advantages over other methodologies, but a number of limitations as well. The two studies in which retrospective reports were directly compared to either prospective or near real time reports yielded mixed results regarding degree of agreement. Findings from several areas of research suggest that some substance abusers may have difficulty recognizing and reporting internal states. CONCLUSIONS: Despite memory problems and other limitations of self-report, all three methodologies have produced findings that implicate negative affect states, increased craving, cognitive factors, interpersonal problems, and lack of coping as factors in relapse. Future relapse studies should make use of all three methodologies to obtain data on relapse and consequences, and steps should be taken to enhance the accuracy of the self-reports that are gathered with each method. PMID- 10463815 TI - Determination of tumor necrosis factor-alpha (TNF-alpha) in serum by a highly sensitive enzyme amplified lanthanide luminescence immunoassay. AB - OBJECTIVES: To develop a highly sensitive enzyme amplified lanthanide luminescence (EALL) immunoassay for tumor necrosis factor-alpha (TNF-alpha). METHODS: The method is based on the use of two monoclonal antibodies against TNF alpha, one "capture" antibody and one labeled with biotin, in a "sandwich type" assay format. Alkaline phosphatase (ALP) conjugated to an antibiotin-polyclonal antibody is used as the enzyme label. ALP cleaves phosphate from diflunisal phosphate (DIFP) to produce diflunisal (DIF). The detection system is based on the combination of enzymatic amplification introduced by ALP and the formation of a highly fluorescent terbium complex that can be monitored by time resolved or conventional fluorimetry. RESULTS: By using 50 microL of sample, the dynamic range of the assay extends up to 2000 ng/L of TNF-alpha, with a detection limit of 1 ng/L, within-run CVs ranging from 3 to 15% and recoveries of 97 +/- 2%. By using 100 microL of sample the dynamic range of the assay extends up to 1000 ng/L of TNF-alpha with a detection limit of 0.2 ng/L, recoveries of 94 +/- 13%, within run CVs ranging from 2 to 6.5% and between-run CVs ranging from 5 to 15%, in a total incubation time of 3h. No interference by the presence of other cytokines (IL-1beta IL-2, IL-4, IL-6, IFN-gamma) or by rheumatoid factors has been observed. The results obtained by the proposed method and by a commercially available kit (Medgenix TNF-alpha EASIA) correlated well (n = 26, r = 0.934). CONCLUSION: The proposed method is highly sensitive, simple and rapid and can reliably measure TNF-alpha in the ng range in biological specimens. PMID- 10463816 TI - Development of an enzyme-linked immunosorbent assay, using a monoclonal antibody against alpha2-macroglobulin, for the diagnosis of systemic lupus erythematosus. AB - OBJECTIVES: To develop an enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody (mab) directed against abnormally glycosylated serum alpha2 macroglobulin (alpha2-M) from patients with systemic lupus erythematosus (SLE). DESIGN AND METHODS: Serum alpha2-M purified by HPLC from patients with SLE was injected in a Balb/c, CB6 F1 female mouse and hybrid cell lines were screened using alpha2-M Glu-C fragments derived from SLE and normal donors (NHS). A mab was selected and used to develop an ELISA by which sera from NHS (n = 14), SLE (n = 34), rheumatoid arthritis (n = 15), Sjogren's syndrome (n = 11), mixed connective tissue diseases (n = 12), and liver diseases (n = 11) were analyzed. RESULTS: The affinity of the mab for alpha2-M from SLE, but not from the other diseases, was higher compared to NHS, as demonstrated by immunoblotting and ELISA. CONCLUSIONS: The ELISA was capable of recognizing changes of glycosylation of alpha2-M in SLE and may be useful for its differential diagnosis. PMID- 10463817 TI - Sandwich enzyme-linked immunosorbent assay for plasma cholesteryl ester transfer protein concentration. AB - OBJECTIVES: Cholesteryl ester transfer protein (CETP) mediates the transfer of HDL cholesterol to apoB-containing lipoproteins. Its mass and activity are increased in several pro-atherogenic conditions. The objective of this study is to develop a cost- and time-effective sandwich ELISA for plasma CETP concentration. DESIGN AND METHODS: Monoclonal anti-CETP, TP20, was used as the capture antibody, while the other biotinylated monoclonal anti-CETP, TP2, was used for detection. The results were expressed in an arbitrary unit, ng biotin TP2 bound per microl plasma. Plasma CETP concentrations, activities and their relationship were assessed in 35 IDDM children. RESULTS: The assay had an intra assay CV of 8.75% and an inter-assay CV under 10%. Plasma CETP concentration of these subjects ranged from 0.36-1.89 ng biotin-TP2/microL. CETP concentration was significantly correlated with CETP activity (r = 0.51, p < 0.01). CONCLUSION: The sandwich ELISA we have developed carried sufficient sensitivity for assaying plasma CETP concentration in human. PMID- 10463818 TI - Measures of total free radical activity in critically ill patients. AB - OBJECTIVES: To examine the role played by free radicals during the initial phase of critical illness in patients on an Intensive Care Unit (ICU). DESIGN AND METHODS: Serum total antioxidant status (TAS) and uric acid (UA) levels were measured in 50 patients over 18 hours to represent the initial stage of critical illness. Clinical scoring systems (APACHE II and multiple organ dysfunction scores) were used to assess the degree of organ dysfunction. Outcome was assessed according to patient survival (survivors, n = 36; non-survivors, n = 14). RESULTS: Serum TAS was higher in non-survivors; a similar finding was demonstrated for serum UA. Levels of both biochemical markers were associated with the degree of organ function and with higher antioxidant and UA levels present in patients with more severe organ dysfunction. In addition, serum UA was significantly correlated to serum TAS and probably accounted for much of the antioxidant activity observed. CONCLUSIONS: The increased TAS and UA levels observed may simply be a response to the degree of renal dysfunction observed as those patients with worse renal function had higher TAS and UA levels. In conclusion, measurement of serum TAS appears to be a reflection of UA concentration and results should be interpreted with caution particularly in patients who have renal dysfunction. PMID- 10463819 TI - Activity of blood coagulation and fibrinolysis during and after hydroxyethyl starch (HES) colloidal volume replacement. AB - OBJECTIVES: To examine the effect of medium molecular weight hydroxyethyl starch on protein C levels and the changes in the activation state of blood platelets, coagulation and fibrinolyis during and after 5 day of its infusion. DESIGN AND METHODS: Fifty male patients (mean age: 47 years, range 45-50 years) who required prostatectomy for benign prostatic hyperplasia were divided into two equal groups. One group was given 15 mL/kg body weight (mean volume 1000 mL +/- 100 mL) of 6% hydroxyethyl starch (HES) 200/0.5, the other received an equal volume of 5% human albumin during the operation. Blood samples were collected immediately before infusion (baseline values) and at 20, 40, 60, 90, 240, and 480 min after the infusion started then daily for the next 5 days postoperatively. Hematocrit, factor VIII:C, thrombin-antithrombin III complex; the anticoagulant protein C levels; the fibrinolytic parameters tissue type plasminogen activator (t-PA), and the fibrinolytic product D-Dimer and the platelet aggregation activity were measured. RESULTS: The data obtained did not detect any significant differences between HES and human albumin in the plasma levels of thrombin-antithrombin III complex, protein C, tissue-type plasminogen activator and the fibrin split products D-Dimer. Factor VIII:C and platelet aggregation were significantly lower in the hydroxyethyl starch group in comparison with albumin. Baseline values were attained postoperatively for factor VIII:C and platelet aggregation by the first and fifth days, respectively. CONCLUSION: The lowering effect of medium molecular weight hydroxyethyl starch on factor VIII:C would not be attributed to increased proteolytic activity of protein C on this coagulation cofactor because there is a nonsignificant change in protein C levels. PMID- 10463820 TI - Relation of genetic polymorphisms of apolipoprotein E, angiotensin converting enzyme, apolipoprotein B-100, and glycoprotein IIIa and early-onset coronary heart disease. AB - OBJECTIVE: Apolipoprotein E (APOE) E4, apolipoprotein B-100 (APOB) Q3611 allele, the angiotensin converting enzyme (ACE) deletion (D) allele and glycoprotein IIIa (GP3A) P33 mutant allele are reported to predispose to early-onset coronary heart disease (CHD). These associations were not all confirmed in more recent studies. To determine the impact of these alleles on CHD, we examined the prevalence of these mutations in patients presenting with early-onset CHD and compared them to those manifesting CHD later in life. The delayed-onset was considered a sign of longevity and would serve as a comparative group to assess prevalence of the biochemical and genetic risk factors. METHODS: 300 patients with a history of myocardial infarction or angina pectoris and angiographically documented CHD were studied. Patients were divided into two groups: group 1 (G1 = 150 patients) presenting with these findings under the age of 50 years; while group 2 (G2 = 150 patients) were patients presenting for the first time over the age of 65 years. Prevalence of the alleles of APOE, APOB, ACE and GP3A was assessed by molecular analysis. An association of any of these genotypes with early onset CHD could lead to a higher prevalence in the younger age group. RESULTS AND CONCLUSIONS: None of the suspected alleles namely APOB Q3611 [G1: 10.7% vs. G2: 9.0%, p = 0.57], ACE D (G1: 52.0% vs. G2: 49.7%, p = 0.57), or the GP3A P33 (G1: 17.3% vs. G2: 15.7%; p = 0.58) showed any significant difference between the two groups. Subjects with APOE E4 were more frequent in the younger age group (G1: 18.3% vs. G2: 13.7%; p = 0.047), while APOE E2 was more frequent in G2 (G2: 10.0% vs. G1: 2.7%; p = 0.0002). Multivariate analysis showed an odds ratio of APOE E2 allele in G1 of 0.27 with a confidence interval of 0.10-0.73. PMID- 10463821 TI - Serum adenosine deaminase activities in lung cancer and mesothelioma. PMID- 10463822 TI - Ligase detection reaction for identification of low abundance mutations. PMID- 10463823 TI - Magnetic resonance imaging of the cervical spine in 27 dogs. AB - The cervical spine of 27 dogs with cervical pain or cervical myelopathy was evaluated using magnetic resonance imaging (MRI). Spin echo T1, T2, and post contrast T1 weighted imaging sequences were obtained with a 0.5 Tesla magnet in 5 dogs and a 1.5 Tesla magnet in the remaining 22 dogs. MRI provided for visualization of the entire cervical spine including the vertebral bodies, intervertebral discs, vertebral canal, and spinal cord. Disorders noted included intervertebral disc degeneration and/or protrusion (12 dogs), intradural extramedullary mass lesions (3 dogs), intradural and extradural nerve root tumors (3 dogs), hydromyelia/syringomyelia (1 dog), intramedullary ring enhancing lesions (1 dog), extradural synovial cysts (1 dog), and extradural compressive lesions (3 dogs). The MRI findings were consistent with surgical findings in 18 dogs that underwent surgery. Magnetic resonance imaging provided a safe, useful non-invasive method of evaluating the cervical spinal cord. PMID- 10463824 TI - Identification of an ossicle associated with the palmar aspect of the carpus in the horse. AB - An ossicle associated with the palmar aspect of the carpus is described in five horses. The size of the ossicle varied from 3 x 4 mm to approximately 12 x 10 mm. The ossicles were located at the junction of the proximal and distal rows of carpal bones, between the palmaromedial aspect of the fourth carpal bone and the ulnar carpal bone. In 3 horses they were bilateral, in one horse unilateral, and only one limb was examined in the remaining horse. In all horses it was an incidental finding and not regarded to be a pathologic entity. PMID- 10463825 TI - Detection of a radiographically occult fracture of the lateral palmar process of the distal phalanx in a horse using computed tomography. AB - A horse with a suspected injury of the distal phalanx was examined using radiography at day two and 19 after the onset of the lameness, with no definite diagnosis. Using computed tomography an incomplete fracture of the lateral wing of the distal phalanx was diagnosed at day 25. Based on computed tomography it was determined that the fracture probably did not enter the joint or involve the palmar cortex throughout its length which were of prognostic importance. The day following the CT examination a new oblique radiographic projection was made. In this radiography which was based on, and never would have been attempted without, the information about the geometric pattern of the fracture achieved by CT examination, a radiolucent area indicative of a wing fracture was seen. This report emphasizes the value of computed tomography in diagnosing a geometric complicated nonarticular oblique fracture of the palmar process of the pedal bone in a horse. PMID- 10463826 TI - Radiographic diagnosis: effect of right vs. left lateral recumbency on myelographic appearance of a lateralized extradural mass. PMID- 10463827 TI - A congenital malformation of the maxilla of a horse. AB - A 1-month-old male American Paint Horse was evaluated for a hard swelling on the right side of the maxillary region. On radiographs there was a large, expansile outpouching of the right maxillary bone between the second and third premolar teeth. Computed tomography further characterized the expansile lesion to have a soft tissue component and to originate in the region of a caudal maxillary tooth. Surgical reconstruction of the defect was unsuccessful and the animal was euthanetized. Based on failure to find histopathologic evidence of a neoplasm or cyst, the diagnosis was a congenital malformation. PMID- 10463828 TI - Crossed renal ectopia with fusion in a cat. AB - In humans, crossed renal ectopia is a rare, usually asymptomatic, congenital malpositioning of the kidneys, involving wandering of one or both embryonic kidneys across the midline and usually fusion with its contralateral partner. This is the first report of crossed renal ectopia in the cat. A 7-year-old, neutered male cat was presented with polyuria, polydipsia, and a palpable abdominal mass. Diagnostic investigations, including clinical pathology, survey and contrast radiography, and ultrasonography, confirmed the presence of an ectopic left kidney fused with an orthotopic right kidney, with concurrent renal failure and hypertension. Many features of crossed renal ectopia in this cat are similar to those described in humans, however the vascular supply, the presence of renal failure, and the axial rotation of the fused kidney are atypical. PMID- 10463829 TI - Chronic renal failure associated with nephrolithiasis, ureterolithiasis, and renal dysplasia in a 2-year-old quarter horse gelding. AB - A 2-year-old quarter horse gelding presented for evaluation of polyuria and polydipsia. Azotemia was detected on serum chemistry profile. Small, misshapen, hyperechoic kidneys with decreased corticomedullary demarcation, hydronephrosis, and a right nephrolith were noted ultrasonographically. The diagnosis of end stage kidney disease and dysplasia was made histopathologically using ultrasound guided biopsy. Two ureteroliths were found in the right ureter via cystoscopy, and a nephrolith was seen in the right kidney at necropsy. Clinical, ultrasonographic, and pathologic features of equine urolithiasis and renal dysplasia are discussed. PMID- 10463830 TI - Radiographic diagnosis--benign bone cyst. PMID- 10463831 TI - Magnetic resonance imaging and ultrasonography in the diagnosis of a malignant peripheral nerve sheath tumor in a dog. AB - A 9-year-old male neutered mixed breed dog had a two-month history of progressive left thoracic limb lameness. There was electromyographic evidence of denervation potentials in all muscles of this limb. In magnetic resonance images a multilobulated, hyperintense mass was visible caudal to the middiaphysis of the left humerus on T-2 weighted images. The mass, which was isointense with surrounding tissue on T1 weighted images, extended proximally towards the brachial plexus. The mass was also visible as a fusiform structure of mixed echogenicity sonographically, although fine-needle aspiration performed at this time was nondiagnostic. A malignant peripheral nerve sheath tumor was diagnosed histopathologically. PMID- 10463832 TI - Ultrasonographic evaluation of the canine shoulder. AB - The aim of this study was to determine the normal ultrasonographic anatomy of the canine shoulder. Fourteen shoulders from 7 clinically normal mid-sized dogs were radiographed and imaged using high frequency ultrasound. Each shoulder was isolated postmortem, and the ultrasonographic and gross anatomy was studied during dissection. The ultrasonographic appearance of the shoulder specimens was similar to that found in the live dogs. Twenty-four shoulders isolated postmortem from 12 variably sized dogs were also used to characterize the normal ultrasound anatomy over a range of sizes. Important anatomic structures that could be consistently evaluated were the biceps tendon and bursa, the bicipital groove surface, the supraspinatous tendon, the infraspinatous tendon, the teres minor tendon, and the caudal aspect of the humeral head. Results of ultrasonographic examination of 4 dogs with shoulder lameness are described to illustrate some applications of canine shoulder ultrasonography in the evaluation of the canine shoulder. In these dogs, ultrasound was a valuable tool to evaluate effusion and synovial proliferation within the bicipital bursa, supraspinatous and biceps tendinitis, biceps tendon strain, and dystrophic calcification. PMID- 10463833 TI - Ultrasonographic evaluation of renal autografts in normal cats. AB - The purpose of this study was to determine changes in resistive index (RI) and renal size that occur in normal feline kidney autografts in the immediate post transplant period. Qualitative changes in renal parenchymal echogenicity and pelvic dilation were also noted, although this was not a major focus of the study. Fourteen domestic short-haired cats underwent bilateral nephrectomies followed by an autograft of the right kidney to either the aorta or external iliac artery. Due to post-operative complications, only nine of these cats completed the study. Ultrasonographic determination of the RI and renal size was made before transplantation and on days 1, 4, 7, 10, and 13 following transplantation. No significant changes in renal RI were identified during the study. Renal size increased significantly, peaking at 63% above baseline on day 7, and stayed elevated at 50% above baseline on day 13. There was no evidence of rejection or other abnormalities found on histologic evaluation of the transplanted kidneys. It was concluded that the RI of normal feline renal autografts does not significantly change in the immediate post-operative period. However, significant renal enlargement, likely due to a combination of post operative edema and hypertrophy, does occur. This study provides baseline information for comparison with feline allografts in the immediate post transplant period. PMID- 10463834 TI - Urocystolith detection: comparison of survey, contrast radiographic and ultrasonographic techniques in an in vitro bladder phantom. AB - Urocystoliths of 9 mineral types from 437 canine patients submitted to the University of Minnesota Urolith Bank were imaged in a urinary bladder phantom. Imaging techniques simulated were survey radiography, pneumocystography, double contrast cystography (two iodine concentrations) and real-time ultrasonography (3.5 MHz, 5.0 MHz, 7.5 MHz). Imaging techniques were compared for accuracy of urocystolith detection, accuracy of urocystolith enumeration, and tendencies for over or undercounting. Across urocystolith mineral types, the false negative rates (no urocystoliths detected in a given case) for survey radiographs range from 2 to 27%. Pneumocystographic techniques are one-half as likely to yield false negative results as are survey radiographic techniques. Underestimates of urocystolith numbers and false negatives are likely using 80 mg iodine/ml double contrast cystography because calcium-based urocystoliths are isopaque in this contrast medium dilution. The 200 mg iodine/ml double contrast cystographic techniques are unlikely to yield false negative diagnoses even for very small (< or = 1.0 mm) urocystoliths and is comparable to pneumocystography for detection and slightly better for enumeration. The likelihood of an ultrasonographic false negative for urocystoliths decreases with increasing MHz. Under optimal conditions using a 7.5 MHz mechanical sector transducer, the false negative rates were comparable to double contrast cystography, but rates increased notably with lower MHz transducers. PMID- 10463835 TI - Spermatic granuloma in the ram: diagnosis by ultrasonography and semen characteristics. AB - Ultrasonographic appearance of the testis and epididymis, and seminal characteristics, with regard to localization of spermatic granuloma were studied. In rams with spermatic granuloma (n = 9), diagnosed by clinical or ultrasonographic examinations before histopathological confirmation, localization of each lesion was recorded. Epididymal granulomas, caput and cauda, were seen as anechoic or hyperechoic areas with a distinct margin with or without a hyperechoic capsule. Granulomas in the testis were microscopic and, therefore, could not be detected by ultrasonography. Enlargement in the mediastinum testis was detected in all rams when there were granulomas in the caput. Heterogeneous testis parenchyma invaded by numerous hyperechoic foci, representing testicular degeneration, was associated with granulomas both in the epididymis and testis. Ultrasonographic appearance of the lesions did not differ with regard to season. Seminal characteristics varied between rams. It was concluded that ultrasonographic evaluation may give valuable information in the diagnosis of sperm granuloma in the ram. PMID- 10463836 TI - Comparison of ultrasonographic and radiographic measurements of canine prostate dimensions. AB - The purposes of this study were to: 1) compare prostatic dimensions measured on radiographic and ultrasonographic images and 2) compare a subjective radiographic assessment of prostate size with a previously-described objective method. Thirty four male dogs undergoing investigation of prostatic disorders were used. Prostate length and depth were measured from ultrasonographic and radiographic images. A subjective assessment of prostate size ('small', 'normal', or 'enlarged') was made in 29 animals by one of the authors who was unaware of radiographic or ultrasonographic measurements. In addition, the distance from sacral promontory to the pubic brim was also measured. A prostate length or depth of >70% of this distance was defined as 'enlarged' and <70% as 'normal'. After the effects of magnification on radiographic measurements were eliminated, there were no significant differences between prostatic length measured by the two methods. However, a significant difference was obtained between prostatic depth measurements. The subjective assessment agreed with a previously described objective assessment of prostatic size in 21/29 dogs for prostate length but in only 12/29 for depth. Prostatic length varied from 46.6 to 116.4% (mean 75.7%) of the distance from the pubic brim to the sacral promontory. Prostatic depth varied from 33.0% to 94.6% (mean 59.7%) of the same distance. It is recommended that prostate length, rather than depth, be used when evaluating prostate size from lateral abdominal radiographs. PMID- 10463837 TI - Effect of transfusion on the endothelium dependent dilatation of brachial artery in patients with chronic anemia. AB - The effect of changes in the main determinants of whole blood viscosity after red blood cell transfusion on endothelium dependent dilatation of brachial artery was studied in patients treated with symptoms of chronic anemia. 10 patients were involved in the study, 8 males, 2 females, mean age 52.7+/-18.7 years. Following blood tests performed at hospital admission: hemoglobin (Hgb), red blood cell count (RBC), hematocrit (Hct), white blood cell count, platelet count, plasma total protein, fibrinogen, plasma viscosity, BUN, creatinine, cholesterol, triglycerides. Flow mediated dilatation of brachial artery (FMD) was determined by Doppler method. Blood tests and FMD study were repeated after transfusion. The main determinants of whole blood viscosity increased after transfusion. The increases of Hgb, RBC, Htc were highly significant. The central flow velocity in brachial artery decreased at rest and during hyperemia as well. FMD of brachial artery was not changed significantly after transfusion. The change in factors determining whole blood viscosity did not cause a significant change in FMD. The probable explanation for this is that the increase of whole blood viscosity is associated with a decrease of central flow velocity. These two counteracting changes can equal each other. PMID- 10463838 TI - The relationship between normal erythrocyte sedimentation rate of Chinese young people and geographical factors. AB - In order to provide a scientific basis for the layout of a unified standard of the reference value of Chinese young people's erythrocyte sedimentation rate (ESR), the relationship between the reference values of the Chinese healthy young's ESR and five geographical factors was tested according to the Wintrobe methods. The altitude was found to be the most important factor affecting the reference value of the Chinese young's ESR. The altitude increase correlates with the decrease of the reference value of the young's ESR. The method of stepwise regression analysis was used to deduce two multivariate regression equations. If the geographical index values of a particular area of China are known, the reference value of the young's ESR of this region can be calculated by means of the regression equations. Furthermore, the depending on geographical factors, China can be divided into six districts: Qingzang, Southwest, Northwest, Southeast, North and Northeast District, according to the reference values of the Chinese young's ESR. PMID- 10463839 TI - Rheological action of aspirin on human erythrocytes. AB - The rheological action of aspirin on human erythrocytes was studied by in vitro incubation of normal erythrocytes with aspirin at 1, 2 and 4 mg/ml for 30-60 min and then measuring erythrocyte rheological properties of deformability, osmotic fragility and aggregation. Aspirin (2 and/or 4 mg/ml) significantly (p<0.05) prevented the loss of filterability (deformability) through 5 microm diameter pores of erythrocytes dehydrated with hypertonic buffer (450 mOsm/kg water) or with potassium ionophore valinomycin (18 micromol/l). When the calcium ionophore A23187 (1.9 micromol/l) was used to induce cell dehydration, aspirin (4 mg/ml) unexpectedly significantly (p<0.05) increased further the loss of filterability (deformability). Aspirin (1, 2 and 4 mg/ml) also increased significantly (p<0.05) erythrocyte osmotic fragility. Aspirin (4 mg/ml) had no effects on aggregation of erythrocytes induced by dextran 70 (3%). The results suggest that aspirin could play a rheologically active role on erythrocytes. The observed effects of aspirin could be explained by acetylation of intracellular proteins and hence saturation (concentration) of the cell interior with the osmotically active drug. PMID- 10463840 TI - Hemorheology of growth hormone-deficient adults. AB - Growth hormone deficiency (GHD) in adults results in alterations of body composition and metabolism associated with a lowered insulin sensitivity and an increased cardiovascular risk. Since hemorheologic disturbances (putative factors of vascular risk) are found in the insulin-resistance syndrome, we investigated blood rheology in 9 adults GHDs (5 men, 4 women; age 37.9+/-4.7 years; body mass index 30.23+/-3.2 kg/m2) compared with 23 lean controls and 37 controls matched for sex, age and body mass index. While this sample of GHDs exhibits the typical metabolic picture of this syndrome (upper body overweight with a waist-to-hip ratio at 0.91+/-0.07; low HDL cholesterol at 1.07+/-0.09 mmol x l(-1); low insulin sensitivity with the minimal model technique at 3.3+/-1.29 min( 1)/(microU/ml) x 10(-4)) they have similar values of blood viscosity at either native or corrected hematocrit, similar hematocrit, similar red cell rigidity viscometric index, similar red cell aggregation parameters than overweight matched controls. There is only a nonsignificant tendency for plasma viscosity to be higher in GHDs: this tendency becomes significant when women are considered alone (GHDs: 1.44+/-0.04 mPa.s; controls: 1.31+/-0.04 mPa.s, p<0.05) while it is no longer found in men. This study suggests that GHDs exhibit the classical hemorheological disturbances of non-GHD individuals with the same degree of obesity. There is no evidence for a further impairment of blood rheology associated with the specific metabolic and endocrine pattern of GHDs that may be involved in their increased vascular risk. PMID- 10463841 TI - Abnormal plasma and serum viscosity in systemic lupus erythematosus (SLE): a Jamaican study. AB - Vascular complications occur frequently in SLE, and their development may be related to haemorheological derangements. However, the range of rheological abnormality in Jamaican SLE patients have not been studied. The present investigation was aimed at determining the changes in the three major blood proteins, namely fibrinogen, albumin and globulin, and their effect on plasma and serum viscosity in SLE patients during the course of treatment. The concentrations of fibrinogen and globulin were significantly increased (p<0.001), while albumin was decreased (p<0.05) in the SLE patients as compared with the control group. This increase in the fibrinogen and globulin contributed significantly to the rise of both serum and plasma viscosity. SLE patients with more severe disease had higher serum viscosity values, suggesting that serum viscosity values may provide an important marker for disease severity. PMID- 10463842 TI - The role of spleen in suppressing the rheological alterations in circulating blood. AB - The role of spleen in maintaining the normal rheological properties of red blood cells (RBC) has been investigated by comparing the time course of RBC deformability assessed by the cell transit analyzer (CTA), after the induction of RBC mechanical alterations, in splenectomized and normal guinea pigs. After the exchange transfusions with glutaraldehyde treated (hardened) RBC, most of these cells were removed from the circulation in the splenectomized animals as well as the animals with intact spleens. However, the CTA could detect the longer existence of a small population of hardened RBC in circulation in the splenectomized animals. Measurement of RBC transit times after the onset of experimental sepsis (cecal ligature-puncture) revealed that, in the splenectomized guinea pigs RBC deformability impairment started earlier, in comparison with the animals with intact spleens. These results suggest that the spleen plays an important role in maintaining the normal rheological properties of the circulating blood, especially in the presence of pathophysiological processes affecting RBC mechanics. PMID- 10463843 TI - Effect of acute hypoxia and hypoxic acclimation on hemorheological behavior in rats with frostbite. AB - It was found that cold injury in rats acclimated to hypoxia was more serious than that in nonacclimated ones. In order to go further into the cause, blood rheological parameters in rats of frostbite at normoxia (FN), frostbite during acute hypoxia (FAH) and frostbite during hypoxia after hypoxic acclimation (FHAC) groups were observed before and after freezing. Before freezing, systemic hematocrit (Hct), blood viscosity, RBC aggregation index and RBC rigidity of FHAC group were higher and RBC deformability index (DI) of FHAC group was lower distinctly than those of FN and FAH groups. After frostbite, the aforesaid changes aggravated prominently, plasma viscosity of FHAC group reduced and was lower than that of FN and FAH groups. The results suggest that the hemorheological behavior is deteriorated in rats acclimated to hypoxia or subjected to freezing, and worsens in hypoxic acclimation rats after freezing. The changes in hemorheological behavior may be one of the causes that would make the frostbite damage more severe in rats acclimated to hypoxia. PMID- 10463844 TI - Hemorheological changes during venous stasis as result of tourniquet application. PMID- 10463845 TI - The effect of vitamin E supplementation on erythrocyte osmotic fragility in diabetic patients. PMID- 10463846 TI - Outcome of temporal lobectomy: expectations and the prediction of perceived success. AB - PURPOSE: This study examined the independent contributions of medical and psychosocial factors to perceived surgical success. We aimed to develop a multidimensional model predictive of perceived surgical outcome. METHODS: Fifty anterior temporal lobectomy (ATL) patients were prospectively assessed, using a formally coded, semistructured clinical interview. This has been routinely administered pre- and post-operatively as part of a larger, nationwide study of Australian ATL patients. The interview covers a broad range of epileptological, psychiatric, neuropsychological and psychosocial issues. Variables from these domains were examined in relation to the patient's perception of surgical success at the 6-month post-operative review. RESULTS: Variables that correlated with success were analysed using principal components analysis and multiple regression. A predictive model of perceived surgical success emerged, which highlighted the multidimensionality of outcome. Independent effects were observed for both medical and psychosocial factors. These included the patients' pre operative expectations of surgery, their post-operative seizure outcome, and affective state. The findings also highlighted the importance of discarding sick role behaviours associated with chronic epilepsy, after surgery. CONCLUSIONS: Traditional outcome measures (seizure frequency, post-operative affect) are significant in the patient's evaluation of surgical success. These traditional measures, however, do not account for the process of psychosocial adjustment surrounding seizure surgery. This process involves two major components: (1) positive anticipation of change prior to surgery, and (2) learning to discard roles associated with chronic epilepsy after surgery. PMID- 10463847 TI - Delineation of cryptogenic Lennox-Gastaut syndrome and myoclonic astatic epilepsy using multiple correspondence analysis. AB - PURPOSE: To distinguish various types of childhood severe cryptogenic/idiopathic generalised epilepsy on the basis of reproducible diagnostic criteria, using multiple correspondence analysis (MCA). METHODS: We applied MCA to a series of 72 children with no evidence of brain damage, starting epilepsy between 1 and 10 years, with two or more types of generalised seizures. We excluded patients with infantile spasms or typical absences. MCA was performed on all clinical and EEG parameters, first throughout follow-up, then restricted to the first year of the disease. RESULTS: When including all follow-up variables, there were three groups: (1) Thirty-seven children with male predominance, familial history of epilepsy, simple febrile convulsions, massive myoclonus, tonic-clonic fits. Outcome was favourable, with no seizures and mildly affected cognitive functions. Interictal EEG showed short sequences of irregular 3-Hz spike-waves. (2) In 18 children, clinical characteristics were similar to those of the first group at the early stage, but 95% exhibited myoclonic status and vibratory tonic seizures, with persisting seizures on follow-up. EEG showed long sequences of generalised irregular spike and slow waves. Those two groups meet the characteristics of childhood onset myoclonic-astatic epilepsy (MAE) with respectively, favourable and unfavourable outcome. (3) Eleven children had later onset, atypical absences, tonic and partial seizures, and no myoclonus, or vibratory tonic seizures. All had mental retardation and persisting seizures. EEG showed long sequences of slow spike-wave activity and half the patients had spike and slow wave foci. These patients met the major characteristics of Lennox-Gastaut syndrome. Initial parameters failed to distinguish the first two groups, but Lennox-Gastaut syndrome (the third group) was distinct from both groups of myoclonic astatic epilepsy from the onset. Within MAE groups combined, clinical and EEG risk factors for mental retardation could be identified. CONCLUSION: It is possible to validate statistically the distinction between discrete epileptic syndromes. Myoclonic astatic epilepsy is therefore distinct from Lennox-Gastaut syndrome, and the distinction appears from the first year of the disorder. PMID- 10463848 TI - Abnormal responses to perforant path stimulation in the dentate gyrus of slices from rats with kainate-induced epilepsy and mossy fiber reorganization. AB - Previous electrophysiological studies have demonstrated that in a subset of hippocampal slices from tissue resected from patients with mesial temporal lobe epilepsy, perforant path stimulation can elicit prolonged negative field potential shifts in the dentate granule cell layer (Masukawa et al., 1989. Brain Res. 493, 168-174; Isokawa and Fried, 1996. Neuroscience 72, 31-37). In this investigation, hippocampal slices were prepared from rats: (1) 2-4 days following kainate treatment, when little or no reorganization of the mossy fibers would be present and (2) 3-13 months after kainate treatment, when mossy fiber reorganization would have occurred. In saline-treated controls, perforant path stimulation typically evoked a single population spike. In contrast, perforant path stimulation could evoke 3-12 population spikes in nearly all slices from kainate-injected rats 2-4 days and 3-13 months after treatment. The majority of slices from kainate-injected rats 3-13 months after treatment had qualitatively similar responses to perforant path stimulation as that observed in slices from kainate-injected rats 2-4 days after treatment. However, in 17% of the slices from kainate-treated rats 3-13 months after treatment (29% of rats), the multiple population spikes were followed by a prolonged negative field-potential shift (duration: 140 ms-1.5 s) with variable superimposed population spike activity. This type of epileptiform activity was only observed in slices with robust Timm's staining in the inner molecular layer and similar responses could also be evoked in these slices with hilar stimulation. Furthermore, pharmacological depression of inhibition by adding the GABA(A) receptor antagonist bicuculline unmasked hilar-evoked prolonged negative field-potential shifts in most slices from kainate-treated rats 3-13 months following treatment, and these slices had robust Timm's staining in the inner molecular layer. Such events were not observed in slices from saline-treated controls or kainate-injected rats 2-4 days after treatment. In conclusion, the prolonged negative field-potential shifts evoked to perforant path stimulation in normal ACSF were associated with mossy fiber reorganization, but the relative contribution of altered inhibition, increased synaptic excitation, or even non-synaptic mechanisms is unknown. PMID- 10463849 TI - Glial cell changes in the white matter in temporal lobe epilepsy. AB - Temporal lobe gliosis and neuronal loss are pathological hallmarks of complex partial seizures. However, the specificity of glial cell changes is not clear. To assess this we studied surgically resected temporal lobes containing either medial temporal sclerosis (MTS) or temporal lobe epilepsy with tumour (TLET) and compared them with idiopathic epilepsy cases and normal controls. We quantitatively assessed glial cell density and mean nuclear volume in the white matter of various temporal gyri and the deep white matter. There was an increase in mean glial cell nuclear volume in MTS and TLET cases in the white matter of superior temporal gyrus, parahippocampal gyrus and deep white matter but not in the white matter of the middle temporal gyrus. In contrast, the densities of glial cells immunopositive for glial fibrillary acidic protein in the MTS and TLET groups were reduced in all white matter regions when compared with the controls. These changes may indicate that glial cells in the white matter have an active role to play in epilepsy pathogenesis. PMID- 10463850 TI - Intrauterine growth in the offspring of epileptic women: a prospective multicenter study. AB - The aim of the present study was to evaluate the risk of intrauterine growth delay in the offspring of epileptic mothers and to quantify the risks of intrauterine exposure to antiepileptic drugs (AEDs). Data concerning 870 newborns, prospectively collected in Canada, Japan and Italy, using the same study design, were pooled and analyzed. The overall proportion of newborns whose body weight (7.8%) or head circumference (11.1%) at birth were below the 10th percentile was not increased. However, logistic regression analysis showed that the risk of small head circumference was significantly higher in Italian than in Japanese (RR 4.2; 95% CI: 2.2-8.0) or Canadian children (RR 2.6; 95% CI: 1.1 6.5), and in children exposed to polytherapy (RR 2.7; 95% CI: 1.2-6.3), phenobarbital (PB) (RR 3.6; 95% CI: 1.4-9.4) and primidone (PRM) (RR 4.5; 95% CI: 1.5-13.8). Country was also the only factor affecting low body weight, with Italian children having a higher risk than Japanese (RR 5.2; 95% CI: 2.6-10.4) or Canadian (RR 8.8; 95% CI: 2.0-38.1) children. Due to the small categories, the influence of AED doses and plasma concentrations was studied for each individual AED, without adjustment for the other potential confounding factors. A clear dose dependent effect was found for PB and PRM in terms of both small head circumference and low body weight, and a concentration-dependent effect for PB in terms of small head circumferences. The size of the difference between the Italian and the other two populations, which is only partially explained by differences in therapeutic regimens, suggests that genetic, environmental and ethnic factors also need to be taken into account when considering possible explanations. PMID- 10463851 TI - Association analysis of a regulatory promoter polymorphism of the PAX-6 gene with idiopathic generalized epilepsy. AB - The PAX-6 gene is a member of the paired-box-containing (PAX) gene family, encoding a transcriptional activator, that plays an important role in the development of the central nervous system. The present association study tested the hypothesis that length variation of a novel regulatory dinucleotide repeat polymorphism in the promoter region of the PAX-6 gene (PAX-6 gene-linked polymorphic region, PAX-6LPR) confers susceptibility to the epileptogenesis of common subtypes of idiopathic generalized epilepsy (IGE). The repeat length of the regulatory dinucleotide repeat polymorphism was assessed in 354 German control subjects and 125 German IGE patients, comprising 70 patients with juvenile myoclonic epilepsy (JME) and 55 patients with an idiopathic absence epilepsy (IAE). The allelic distribution of the PAX-6LPR did not deviate significantly between the controls and the IGE patients (Wilcoxon Rank-Sum test: P > 0.76), or both subgroups of either JME patients (P > 0.78) or IAE patients (P > 0.87). Our results do not provide evidence that length variation of the polymorphic dinucleotide sequence in the PAX-6LPR contributes a frequent and relevant effect to the pathogenesis of common subtypes of IGE. PMID- 10463852 TI - Chronic electrode implantation entails epileptiform field potentials in rat hippocampal slices, similarly to amygdala kindling. AB - Evoked field potentials were recorded in the CA3, CA1 and dentate gyrus (DG) of hippocampal slices from amygdala kindled, non-stimulated amygdala electrode implanted, and non-implanted age-matched rats to evaluate the consequences on hippocampal neuronal networks of kindling stimulation versus electrode implantation. No overt modification of field potentials was detected in either the CA1 or the DG areas. In contrast, a very significant increase in the occurrence of repetitive population spikes evoked by single stimuli was detected in the CA3 area in slices from both amygdala kindled and non-stimulated amygdala implanted rats. The epileptiform pattern of CA3 field potentials was at least as well expressed in implanted non-stimulated, as in kindled rats, suggesting that electrode implantation has a major contribution to this marker of epileptogenesis. PMID- 10463853 TI - Cisternal S100 protein and neuron-specific enolase are elevated and site-specific markers in intractable temporal lobe epilepsy. AB - In the brain, S100 protein and neuron-specific enolase (NSE) are mainly found in glial cells and neurons, respectively. We investigated concentrations of S100 protein and NSE in cisternal cerebrospinal fluid obtained during implantation of foramen ovale electrodes in eight patients with temporal lobe epilepsy (TLE). In addition, the meningeal markers cystatin-C and beta-trace as well as total protein were measured. Patients with trigeminal neuralgia (TN) undergoing glycerol rhizotomy served as controls. S100 protein and NSE levels ipsilateral to the site of seizure onset were significantly higher than in TN. Contralateral TLE values were also markedly but not significantly elevated. The meningeal markers cystatin-C and beta-trace protein as well as total protein did not differ in TLE and TN. We conclude that interictal temporal lobe dysfunction corresponds with neuronal and glial marker elevations in the extracellular space and that site specific elevations may predict the site of seizure origin biochemically. PMID- 10463854 TI - Edward Peirson Richardson Jr (1918-1998) and the discovery of PML. PMID- 10463855 TI - Review: neuronal injury in HIV infection. PMID- 10463856 TI - Astrocytes as targets for Venezuelan equine encephalitis virus infection. AB - Venezuelan equine encephalitis virus (VEE) produces an acute infection in humans and induces a well-characterized cytopathic effect in neurons of the central nervous system (CNS). However, little is known about the role of glial cells in response to VEE infection of the CNS. Our results demonstrate that VEE is capable of a productive infection in primary astrocyte cultures and that this infection is cytotoxic. Further, there were significant differences in the growth kinetics comparing virulent and attenuated strains of VEE. Additionally, VEE infection of astrocyte cultures induced gene expression of two neuro-immune modulators, tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS). Assays for TNF-alpha protein and nitric oxide (NO) demonstrated high levels of TNF-alpha protein and low levels of NO in response to VEE infection of astrocytes. These observations suggest an important role of astrocytes in this virus-induced encephalitis, and that interactions between astrocytes, other glial cells, and neurons may be important in VEE pathogenesis. Such interactions, which could impact neuronal survival, may include loss of functional changes in astrocytes or, alternatively, their production of neurotoxic molecules. PMID- 10463857 TI - Selective targeting of habenular, thalamic midline and monoaminergic brainstem neurons by neurotropic influenza A virus in mice. AB - Infections caused by influenza A virus have been proposed to be associated with neuropsychiatric complications, the mechanisms of which remain to be unravelled. We here report that a neurotropic strain of influenza A virus (A/WSN/33) introduced into the olfactory bulbs of C57BL/6 (B6) mice, selectively attacks habenular, paraventricular thalamic, and brainstem monoaminergic neurons. In the habenular and paraventricular thalamic areas, infection was followed by an almost total loss of neurons within 12 days. In the brain stem monoaminergic areas, viral gene products were eliminated from neurons by 12 days in B6 wildtype mice, but remained for at least 35 days in immunodefective TAP1 (Transporter associated with Antigen Presentation 1) mutant mice. In conclusion, we show that influenza A virus infection in the brain selectively targets regions which have been implicated in neuropsychiatric disturbances, and that this virus can remain for a significant period of time in specific regions of the brain in immunodefective mice. PMID- 10463858 TI - Association of human herpesvirus 6 with the demyelinative lesions of progressive multifocal leukoencephalopathy. AB - Progressive Multifocal Leukoencephalopathy (PML) is a primary demyelinating disease of the central nervous system occurring almost exclusively in individuals with impaired cell-mediated immunity. The JC polyoma virus has been accepted as the etiologic agent ofPML. Using a two-step in-situ polymerase chain reaction procedure to amplify and detect genomic DNA of human herpesvirus-6 (HHV6) in formalin-fixed paraffin-embedded archival brain tissues, a high frequency of infected cells was consistently detected in PML white matter both within and surrounding demyelinative lesions and HHV6 genome was found mainly within oligodendrocytes. Lesser amounts of HHV6 genome were detected in most normal, AIDS, and other neurological disease control tissues. Immunocytochemistry for HHV6 antigens showed actively infected nuclei of swollen oligodendrocytic morphology only within the demyelinative lesions of PML but not in adjacent uninvolved tissue. In addition, no HHV6 antigens were detectable in control tissues including brains of individuals with HIV-1 encephalopathy but without PML. Double immunohistochemical staining for JC virus large T antigen and HHV6 antigens demonstrated co-labeling of many swollen intralesional oligodendrocytes in the PML cases. The evidence suggests that HHV6 activation in conjunction with JC virus infection is associated with the demyelinative lesions of PML. PMID- 10463859 TI - Heat stress activates production of herpes simplex virus type 1 from quiescently infected neurally differentiated PC12 cells. AB - We have previously described a novel in vitro model of a non-productive herpes simplex virus type 1 (HSV-1) infection in neurally differentiated (Nd)-PC12 cells that allows for inducible virus replication upon forskolin treatment. In this study, we further characterized the quiescent state of infection and examined the ability of heat stress (HS) to induce virus from this non-productive state. These studies demonstrated that (i) the quiescent state is characterized by the absence of cell-associated virus, capsids, and viral antigens; (ii) HS (43 degrees C, 3 h) efficiently activated virus from quiescently infected Nd-PC12 (QIF-PC12) cells; (iii) the rate of virus production was significantly greater following HS than forskolin treatment, and the rates of both were dependent on MOI; (iv) forskolin and HS appeared to affect pathways of viral activation from a quiescent state as they did not enhance viral growth in Nd-PC12 cells; (v) viral alpha4 gene and host HSP72 gene transcription were rapidly induced in QIF-PC12 as soon as 3 h post-HS initiation; (vi) induction of the viral alpha27 gene followed that of representative beta and gamma genes, U(L)30 and U(L)18, respectively, and (vii) HS induced asynchronous HSV-1 replication from QIF-PC12 cells with 1:400 to 1:22000 positive foci detected as rapid as 24 h post-induction when established at MOIs of 30 and 3, respectively. These findings provide evidence that alpha4 may be involved in the switch from quiescence to productive infection. Furthermore, this model has the potential to advance our understanding of how HS initiates the HSV-1 productive cycle from a cryptic viral genome. PMID- 10463860 TI - Restricted replication of herpes simplex virus in satellite glial cell cultures clonally derived from adult mice. AB - To determine the possible influence of satellite glial cells on restricting the spread of herpes simplex virus in the peripheral nervous system, HSV replication was studied in clonally derived cultures of satellite glial cells from adult animals. Satellite cells were purified by exploiting their close anatomical association with primary sensory neurons. Dissociated neurons from dorsal root ganglia were micro-manipulated to remove all but one of the attached satellite cells and cultured in the presence of the mitogenic stimulators bovine pituitary extract and cholera toxin. Following a lag phase of 20-30 days some of the individual satellite cells began to proliferate. Initial cultures demonstrated bipolar morphology similar to cultured Schwann cells, some of which differentiated into large astrocytic whorl-like cells on subsequent passage. Immunocytochemical and molecular studies demonstrated that these cells, designated Sat.1, express glial fibrillary acidic protein, confirming their glial origin and by electron microscopy they were shown to be phagocytic. Under single step viral growth conditions Sat.1 cells were restrictive for HSV replication, producing in the order of 1000 times less infectious virus than Vero cells, a standard permissive cell line. These results suggest that satellite cells, which tightly encase sensory neurons, play a role in restricting interneural spread of HSV within the peripheral nervous system. PMID- 10463861 TI - Aurothiolates enhance the replication of Semliki Forest virus in the CNS and the exocrine pancreas. AB - The A7(74) strain of Semliki Forest virus (SFV) is avirulent and the L10 strain virulent in adult mice. A7(74) infection of adult mouse brain gives rise to small discrete foci of infection which, in immunocompetent animals, are cleared within 10 days. In contrast L10 infection results in a widespread and fatal central nervous system infection. Aurothiolates are linear, 2-coordinate complexes in which two ligands are covalently bound on either side of a gold nucleus in a +1 oxidation state (gold (I)). Pretreatment of A7(74) infected mice with two distinct aurothiolates (sodium aurothiomalate and aurothioglucose) resulted in significantly increased brain virus titers, and large confluent areas of infection in the brain similar to the pattern of infection seen with the L10 strain. The gold (I) moiety of aurothiolates was demonstrated to be the active component, since thiomalic acid when administered alone had no potentiating effect on the infection. Although both aurothiolates allowed productive replication and spread of A7(74) within the nature mouse brain, enhanced neuronal destruction was not apparent. There were no significant changes in virus distribution in any other tissue except for the exocrine pancreas and the myocardium where widespread infection of the acinar cells and occasional infected myocytes were observed. PMID- 10463862 TI - Immune surveillance and antigen conformation determines humoral immune response to the prion protein immunogen. AB - Transmissible spongiform encephalopathies (TSE) are progressive degenerative disorders of the central nervous system. PrP(Sc) is a TSE-specific marker derived from the host-encoded glycoprotein, PrPc. The generation of antibodies to PrP plays an important role in the diagnosis of these diseases. In this study the role of the PrP immunogen and the species being immunized was examined in relation to specific epitopes. Various mammals (mice, hamsters, rabbits and PrP null mice) were immunized with formic acid-treated PrP(Sc) isolated from mice, hamsters and sheep. Both the species being immunized and the source of immunogen played an important role in the antibody response. Response to a limited number of linear epitopes was seen among the various immunized animals. One region in the C-terminal portion of PrP appeared highly immunogenic in all species. Comparison of immunoreactivity and the pepscan-defined linear epitope sites suggests both linear and conformational directed responses in many of the animals. Information on the forces directing immune responses to PrP will lead to a better understanding of host-PrP interactions. It will also assist in the development of new strategies for generating additional tools for immunodiagnosis. PMID- 10463863 TI - High incidence of meningeal infiltration by leukemic cells after infection of chimeric virus between neuropathogenic and non-neuropathogenic retroviruses. AB - Friend murine leukemia virus (F-MuLV) clone A8, previously shown to cause spongiform degeneration in rat brain, induced leukemia within 10 weeks after infection when inoculated into newborn rats. The chimeric virus Rec2, which contains the pol and env genes of 57 virus on the background of A8 and does not cause spongiform degeneration in the central nervous system (CNS), induced leukemic cell infiltration of the CNS, mainly of the meningeal region, in 58.3% of infected rats. In contrast, A8 induced little or no leukemic cell infiltration of the CNS. Other chimeric viruses containing the LTR and the 5' half of the 5' leader sequence of A8 induced aggressive leukemia, and after infection of these viruses, leukemic cell infiltration of the CNS was only observed in less than 20.0% of the rats. These results indicate that the fragment containing the LTR and the 5' half of the 5' leader sequence of A8 is essential for induction of aggressive leukemia in rats but is not sufficient to cause CNS infiltration. We found that leukemic cell infiltration of the CNS is dependent on the sequence of the virus. PMID- 10463864 TI - Prolonged survival without neurological improvement in patients with AIDS-related progressive multifocal leukoencephalopathy on potent combined antiretroviral therapy. AB - To evaluate the benefit of combined antiretroviral therapy including protease inhibitors (CART) on survival time and neurological progression in patients with AIDS-related progressive multifocal leukoencephalopathy (PML), 81 consecutive PML cases, collected between January 1990 and June 1998, were reviewed. Fifteen patients were neuropathologically proven. JC virus detection in CSF was positive in 59 patients. At PML diagnosis, median CD4 cell count was low (median, 35 cells/microL) and plasma HIV load, determined in 41 patients, was high (median, 4.8 log10 copies/ml). Following PML diagnosis, there was a significant difference (P<10(-4)) in survival between patients who were untreated or treated with nucleoside analogs (n=50, median: 80 days) and patients who were started early on CART (n=23, median: 246 days). A third group of eight patients who received CART late during the course of PML was considered separately. At the study endpoint, 18 of all the CART-treated patients (n=31) were still alive. Plasma HIV load was undetectable in 67% of them. The median increase in CD4 cell count was 112 cells/microL from CART onset. In contrast, no significant improvement in neurological status was observed. Our results demonstrate a benefit of CART on survival of AIDS-related PML patients and suggest the need for an early, specific anti-JC virus treatment to limit the neurological deterioration. PMID- 10463865 TI - Detection of JC virus DNA in peripheral blood cell subpopulations of HIV-1 infected individuals. AB - While it has been suggested that JC virus (JCV) migrates in B-lymphocytes from the kidney to the central nervous system where it initiates demyelination, this phase of JCV pathogenesis has not been systematically explored. To determine the peripheral blood cell subpopulation(s) infected with JCV, monocytes, granulocytes, and T and B lymphocytes from HIV-1-infected individuals and uninfected controls were purified by flow cytometry. JCV DNA could be detected by PCR amplification in all of these cell subpopulations. This finding suggests that JCV lacks specificity in its interaction with leukocytes. PMID- 10463866 TI - The applied neuropathology of human spinal cord injury. PMID- 10463867 TI - Late-onset, bleb-associated endophthalmitis following glaucoma filtering surgery with or without antifibrotic agents. AB - The use of adjunctive antimetabolites such as 5-fluorouracil (5-FU) and mitomycin C (MMC) in trabeculectomy has caused an increased concern as to the risk of bleb associated endophthalmitis. In this study, we compared the incidence of this complication between different methods of adjunctive treatment in glaucoma filtering surgery. Records of 15 cases of late-onset, bleb-associated endophthalmitis which were admitted to the Ophthalmology Department, Hadassah University Hospital, between January 1980 and December 1997 were retrospectively studied. These patients had endophthalmitis at least two months following glaucoma filtering surgery. Bleb-associated endophthalmitis occurred in 2 eyes following filtering surgery with no antifibrotic agent, in 6 eyes following trabeculectomy with 5-FU, and in 7 eyes that underwent trabeculectomy with intraoperative MMC. The mean time from surgery to endophthalmitis following 5-FU was 25.0+/-22.0 months (range 3 to 56 months), and 11.3+/-5.4 months (range 4 to 18 months) following the use of MMC. The estimated cumulative incidences of bleb associated endophthalmitis in superior blebs at 24 months following surgery were 0.3% for surgeries without antifibrotic agents, 0.8% for trabeculectomies with postoperative 5-FU, and 1.3% for trabeculectomies with intraoperative MMC. No significant differences were demonstrated between the survival distributions of filtering procedures performed with any of the three techniques (p = 0.166, Log rank test). Most of the cases following 5-FU or MMC appeared within 18 months following trabeculectomy. The visual outcome of most of the eyes was poor. Antifibrotic agents are not associated with a higher risk for late-onset, bleb associated endophthalmitis. A high rate of alertness is warranted for diagnosis of early signs of infection following glaucoma filtering surgery. PMID- 10463868 TI - Role of nitric oxide in mediating retinal blood flow regulation in cats. AB - The role of nitric oxide (NO) was evaluated in mediating control of retinal blood flow in the anesthetized cat. This was done under resting conditions and as a function of decreases in perfusion pressure. The retinal blood flow was calculated by measuring blood velocity with laser Doppler velocimetry (LDV) and retinal blood vessel diameter with image analysis. Graded decreases in perfusion pressure were obtained by increasing intraocular pressure (IOP). Following intravenous administration of 30 mg/kg NG-nitro-L-arginine methyl ester (L-NAME), an NO synthase inhibitor, the arterial blood pressure increased under resting conditions by 34+/-3% (n = 7), the blood vessel diameter decreased by 18+/-2%, and retinal blood flow significantly declined by 27+/-6% (n = 6). Irrespective of the presence or absence of L-NAME (30 mg/kg), stepwise reductions in perfusion pressure that reached a level that was 35% of the baseline value had no significant effect on retinal blood flow. These results suggest that, under resting conditions, retinal blood flow decreases in response to a putative fall in NO levels. However, NO does not appear to be involved in mediating the autoregulatory response to a decrease in perfusion pressure. PMID- 10463869 TI - The effectiveness and safety of dorzolamide 2% in addition to multiple topical antiglaucoma medications. AB - Dorzolamide is the first commercial topical carbonic anhydrase inhibitor approved by the Food and Drug Administration (FDA) for the treatment of glaucoma. In a prospective, open label, uncontrolled study on 245 glaucoma patients, dorzolamide significantly lowered the intraocular pressure at least 14% when used alone or in combination with one, two, or three other antiglaucoma medications over ten weeks. There were very few adverse reactions to dorzolamide. Dorzolamide is effective and safe when used alone or in combination with other topical antiglaucoma medications for the treatment of glaucoma. PMID- 10463870 TI - Effect of topical betaxolol on tissue circulation in the human optic nerve head. AB - There have been no reports to date on long-term betaxolol instillation effects on the human optic nerve head (ONH) tissue circulation. The purpose of this study was to study the effect of topical 0.5% betaxolol on tissue blood velocity in the human ONH. Using a laser-speckle tissue blood flow analyzer, normalized blur (NB; a quantitative index of tissue blood velocity) was measured every 0.125 seconds at a temporal ONH site free of visible surface vessels. Measurements were averaged for 3 cardiac cycles (NB(ONH)). For baseline comparison (day 0), recordings of bilateral NB(ONH) and intraocular pressure (IOP), blood pressure (BP) and pulse rate (PR) were recorded in healthy volunteers before, and 2, 4.5, and 7 hr after, instillation of 30 microL of betaxolol vehicle, and again on day 21; IOP was also recorded on days 7 and 14. On day 1 (the day after baseline measurements), and twice daily for 3 weeks, 30 microL of 0.5% betaxolol into one eye and 30 microL vehicle was instilled into the other in a double-blind study. Measurements as on day 0 were again recorded on day 21; IOP was also recorded on days 7 and 14. During baseline recordings, no significant changes were noted in any parameters. After administration of topical betaxolol, IOP was significantly reduced, bilaterally, with greater reduction in the betaxolol-treated eyes on day 21. Also on day 21, the NB(ONH) of the betaxolol-treated eyes was significantly higher 4.5 hr after instillation than that of the comparable baseline recording (p = 0.035 with Bonferroni's correction); BP, PR, and NB(ONH) in the eye which received only the vehicle showed little change. Tissue blood velocity in the human ONH was increased at least temporarily by instillation of topical betaxolol twice daily for 3 weeks. Although the obtained increase is small and may be clinically insignificant, the potential of betaxolol that can affect the ONH tissue circulation in humans after 21 days of instillation is thought to deserve further investigation. PMID- 10463871 TI - Quantitative autoradiographic visualization and pharmacology of FP-prostaglandin receptors in human eyes using the novel phosphor-imaging technology. AB - Quantitative autoradiographic visualization of FP prostaglandin receptors was determined in postmortem human eye sections using [3H]PGF2alpha and the novel phosphor-imaging technology coupled with computerized image analysis. Densitometric analysis of digital images obtained after a 19-day screen-exposure revealed FP receptors to be highly concentrated in the longitudinal ciliary muscle (12,741 digital light units [DLU/mm2]), iris sphincter muscle (19,261 DLU/mm2) and retina (9,544 DLU/mm2), with lesser amounts (1,558-3,776 DLU/mm2) in five other ocular structures (n = 3-4 donors). The highest percentage specific binding was in the iris sphincter and longitudinal ciliary muscle (78-73%) and lowest in the lens (12%). Binding of [3H]PGF2alpha to the longitudinal ciliary and iris sphincter muscles and retina was concentration-dependently and potently displaced by known FP-receptor-selective compounds such as cloprostenol (Ki = 13 37 nM), fluprostenol (Ki = 56-98 nM), PHXA85 (Ki = 181-206 nM) and latanoprost (isopropyl ester of PHXA85; Ki = 0.43-4.5 microM) (from up to 4 donor eyes). These quantitative phosphor-imaging autoradiography data provide further evidence for the presence of FP receptors in human longitudinal ciliary muscle, one of the tissues involved in the intraocular pressure lowering effects of FP-class prostaglandins. The quantitative localization of FP receptors in the human iris, iris sphincter muscle, and retina represent interesting new observations. PMID- 10463872 TI - Effects of 0.2% brimonidine on ocular anterior structures. AB - The aim of the study was to evaluate the effect of an alpha2-adrenergic agonist, 0.2% brimonidine, on a number of echobiometry and ultrasound biomicroscopy ocular parameters. Ten healthy subjects ranging in age from 20 to 40 years (mean age 29+/-3.39) were recruited into this prospective open-label trial. After instillation of 0.2% brimonidine eye drops, the following parameters were assessed: refraction, visual acuity, pupil diameter, intraocular pressure, five A scan echobiometric parameters and 15 ultrasound biomicroscopy parameters. As early as the first hour after administration of the drug, a marked ocular hypotensive effect was detected associated with a miotic effect, without any refractive or visual acuity alterations. The A-scan echobiometry parameters were unchanged, while, as far as the ultrasound biomicroscopy variables were concerned, we observed an increase in iris-lens contact distance and a reduction in iris root thickness with a resulting increase in posterior chamber depth and in iris-ciliary process distances. No changes were observed in either the anterior chamber or the anterior iris profile. Brimonidine 0.2% proved to be an ocular hypotensive agent which is also endowed with a mild miotic effect, without giving rise to refractive or visual acuity alterations. The drug does not alter the thickness and position of the lens and does not facilitate pupil block; it reduces the iris thickness with an increase in posterior chamber depth and in iris-ciliary process distance but with no changes in anterior chamber depth or chamber angle width. PMID- 10463873 TI - Effect of vitamin E eye drops on naphthalene-induced cataract in rats. AB - The effect of vitamin E acetate (VEA) eye drops on naphthalene-induced cataract in rats was investigated by Scheimpflug image analysis. The control group was administered only naphthalene (1 g/kg), while the other group was additionally given 1% VEA eye drops into both eyes 5 times a day every day for 9 weeks from the start of naphthalene treatment. During those 9 weeks, the changes of the crystalline lens were documented by an anterior eye segment analysis system (EAS 1000, NIDEK) once a week in mydriasis (Mydrin-P, Santen Pharmaceutical Co., Ltd.). The characteristic density values of the anterior deeper cortex regions were measured. The light scattering intensity of lenses from VEA eye drop-treated animals was significantly lower than that of animals without VEA treatment. This difference was found 1 week, 4 weeks, and from 7 to 9 weeks after the start of naphthalene application. VEA eye drops may have the potential to delay the progression of naphthalene-induced cataract in rats. PMID- 10463874 TI - Suppression of interleukin-1alpha-induced uveitis and inhibition of fibroblast like cell proliferation by synthetic interleukin-1 blockers. AB - To treat uveitis and prolong the functional life of filtration surgery on glaucomatous eyes, some interleukin-1 (IL-1) blockers were used to inhibit IL-1 induced uveitis in rat eyes and to suppress proliferation of fibroblast-like corneal and conjunctival cells in the cell cultures, respectively. It was found in this research that the blood-aqueous barrier can be broken by IL-1alpha to allow fluorescein to enter the eyeballs and to be detected by fluorotron. The uveitis was effectively blocked by the IL-1 blockers studied in this research. It was also noted that the proliferation of fibroblast-like corneal and conjunctival cells was effectively inhibited by IL-1 blockers. The inhibition of cell growth seems to be caused primarily by the inhibition of RNA synthesis. There was a significant difference in the potency of IL-1 blockers to inhibit corneal vs. conjunctival cells. It was noted that conjunctival cells were more easily inhibited by IL-1 blockers than corneal cells. These results indicate that IL-1 blockers can suppress the proliferation of conjunctival cells at dose levels which do not affect the normal cell growth of corneal cells. PMID- 10463875 TI - Intravitreal toxicology and therapeutic efficacy of the carboxymethyl ester of the 1-O-octadecyl-sn-glycerol-3-phosphonoformate (ODG-PFA-O-Me), a novel lipid antiviral prodrug for intraocular drug delivery. AB - This study was conducted to evaluate the vitreous clarity and intraocular therapeutic index of three preparations ofthe carboxymethyl ester of 1-O octadecyl-sn-glycerol-3-phosphonoformate (ODG-PFA-O-Me), a long acting lipid derivative of foscarnet with potent anti-CMV activity. Twenty-six New Zealand white rabbits were intravitreally injected with one of three preparations of ODG PFA-O-Me or control diluent. The vitreous clarity was graded after injection using indirect ophthalmoscopy and fundus photography. Drug intraocular toxicity was evaluated by electroretinography and by post-sacrifice tissue pathology using light and electron microscopy. Intravitreal injection of micellar ODG-PFA-O-Me showed variable local retinal toxicity and vitreal compound aggregates in eyes with the middle and high doses. The intraocular therapeutic index was lower than 465:1. Intravitreal injection of liposomal ODG-PFA-O-Me, either free acid or sodium salt, revealed clear vitreous for the 0.632 and 0.84 mM final intravitreal concentrations. No retinal toxicity was confirmed for the 1.12 mM final intravitreal concentration at the eight week observation following injection. The intraocular therapeutic index was between 585-1037:1. ODG-PFA-O-Me possesses better vitreous compatibility than ODG-PFA. Liposomal ODG-PFA-O-Me can be intravitreally injected with a resulting clear vitreous and high intraocular therapeutic index. Liposomal ODG-PFA-O-Me could be a long acting nontoxic intravitreous injectable drug for CMV retinitis. PMID- 10463876 TI - Managing patients with lung cancer. New guidelines should improve standards of care. PMID- 10463877 TI - Learning from differences within the NHS. Clinical indicators should be used to learn, not to judge. PMID- 10463878 TI - Epidural anaesthesia and analgesia: better outcome after major surgery?. Growing evidence suggests so. PMID- 10463880 TI - Study gives reassurance on safety of xenotransplantation PMID- 10463879 TI - Hereditary haemochromatosis: to screen or not. Conditions for screening are not yet fulfilled. PMID- 10463881 TI - Hypothyroidism in pregnancy affects babies' intelligence PMID- 10463882 TI - Canadian doctors' threatened walk out averted PMID- 10463883 TI - Junior doctors step up campaign over pay PMID- 10463884 TI - In brief PMID- 10463885 TI - FDA bans blood donation by people who have lived in UK. PMID- 10463886 TI - Hepatitis C infects almost 2%of US population PMID- 10463887 TI - Report calls for elimination of tuberculosis in the US by 2010. PMID- 10463889 TI - German doctors are unhappy about drugs budget PMID- 10463888 TI - Overuse of animal antibiotics threatens human health. PMID- 10463890 TI - Guidelines ignored on resuscitation decisions PMID- 10463892 TI - Intercalated degrees, learning styles, and career preferences: prospective longitudinal study of UK medical students. AB - OBJECTIVES: To assess the effects of taking an intercalated degree (BSc) on the study habits and learning styles of medical students and on their interest in a career in medical research. DESIGN: Longitudinal questionnaire study of medical students at application to medical school and in their final year. SETTING: All UK medical schools. PARTICIPANTS: 6901 medical school applicants for admission in 1991 were studied in the autumn of 1990. 3333 entered medical school in 1991 or 1992, and 2695 who were due to qualify in 1996 or 1997 were studied 3 months before the end of their clinical course. Response rates were 92% for applicants and 56% for final year students. MAIN OUTCOME MEASURES: Study habits (surface, deep, and strategic learning style) and interest in different medical careers, including medical research. Identical questions were used at time of application and in final year. RESULTS: Students who had taken an intercalated degree had higher deep and strategic learning scores than at application to medical school. Those with highest degree classes had higher strategic and deep learning scores and lower surface learning scores. Students taking intercalated degrees showed greater interest in careers in medical research and laboratory medicine and less interest in general practice than their peers. The effects of the course on interest in medical research and learning styles were independent. The effect of the intercalated degree was greatest in schools where relatively few students took intercalated degrees. CONCLUSIONS: Intercalated degrees result in a greater interest in research careers and higher deep and strategic learning scores. However, the effects are much reduced in schools where most students intercalate a degree. Introduction of intercalated degrees for all medical students without sufficient resources may not therefore achieve its expected effects. PMID- 10463893 TI - Training PMID- 10463891 TI - Familial clustering of Helicobacter pylori infection: population based study. AB - OBJECTIVES: To assess the rate of intrafamilial transmission of Helicobacter pylori infection in the general population and the role of a family's social background. DESIGN: Population survey. SETTING: Campogalliano, a town in northern Italy with about 5000 residents. PARTICIPANTS: 3289 residents, accounting for 416 families. MAIN OUTCOME MEASURES: Prevalence of H pylori infection assessed by presence of IgG antibodies to H pylori. RESULTS: The overall prevalence of H pylori infection was 58%. Children belonging to families with both parents infected had a significantly higher prevalence of H pylori infection (44%) than children from families with only one (30%) or no parents (21%) infected (P<0.001). Multivariate analyses confirmed that children with both parents positive had double the risk of being infected by H pylori than those from families in which both parents were negative. Family social status was independently related to infection in children, with those from blue collar or farming families showing an increased risk of infection compared with children of white collars workers (odds ratio 2.02, 95% confidence interval 1.16 to 3.49). CONCLUSIONS: H pylori infection clusters within families belonging to the same population. Social status may also be a risk factor. This suggests either a person to person transmission or a common source of exposure for H pylori infection. PMID- 10463894 TI - Responses from pharmaceutical companies to doctors' requests for more drug information in Pakistan: postal survey. PMID- 10463895 TI - Severe periconceptional life events and the sex ratio in offspring: follow up study based on five national registers. PMID- 10463896 TI - Fatal erythroderma associated with pentostatin. PMID- 10463897 TI - Heartburn treatment in primary care: randomised, double blind study for 8 weeks. AB - OBJECTIVE: To compare the effects and tolerability of omeprazole and cisapride with that of placebo for control of heartburn in primary care patients. DESIGN: Randomised, double blind, placebo controlled study. SETTING: 65 primary care practices in Norway. PARTICIPANTS: 483 untreated patients with complaints of heartburn >/=3 days a week, with at most grade 1 reflux oesophagitis. INTERVENTIONS: Omeprazole 20 mg once daily, cisapride 20 mg twice daily, or placebo for 8 weeks. MAIN OUTCOME MEASURES: Adequate control of heartburn, defined as 0.001). These results support the hypothesis that homocysteine may affect NC and NT development by its ability to inhibit the NMDAR. One potentially important consequence of this putative mechanism is that homocysteine may interact synergistically with other NMDAR antagonists to enhance its effect on development. PMID- 10463944 TI - Mitochondrial DNA inherited variants are associated with successful aging and longevity in humans. AB - Mitochondrial DNA (mtDNA) is characterized by high variability, maternal inheritance, and absence of recombination. Studies of human populations have revealed ancestral associated polymorphisms whose combination defines groups of mtDNA types (haplogroups) that are currently used to reconstruct human evolution lineages. We used such inherited mtDNA markers to compare mtDNA population pools between a sample of individuals selected for successful aging and longevity (212 subjects older than 100 years and in good clinical condition) and a sample of 275 younger individuals (median age 38 years) carefully matched as to sex and geographic origin (northern and southern Italy). All nine haplogroups that are typical of Europeans were found in both samples, but male centenarians emerged in northern Italy as a particular sample: 1) mtDNA haplogroup frequency distribution was different between centenarians and younger individuals (P=0.017 by permutation tests); and 2) the frequency of the J haplogroup was notably higher in centenarians than in younger individuals (P=0.0052 by Fisher exact test). Since haplogroups are defined on the basis of inherited variants, these data show that mtDNA inherited variability could play a role in successful aging and longevity. PMID- 10463945 TI - Melatonin inhibits expression of the inducible NO synthase II in liver and lung and prevents endotoxemia in lipopolysaccharide-induced multiple organ dysfunction syndrome in rats. AB - We evaluated the role of melatonin in endotoxemia caused by lipopolysaccharide (LPS) in unanesthetized rats. The expression of inducible isoform of nitric oxide synthase (iNOS) and the increase in the oxidative stress seem to be responsible for the failure of lungs, liver, and kidneys in endotoxemia. Bacterial LPS (10 mg/kg b. w) was i.v. injected 6 h before rats were killed and melatonin (10-60 mg/kg b.w.) was i.p. injected before and/or after LPS. Endotoxemia was associated with a significant rise in the serum levels of aspartate and alanine aminotransferases, gamma-glutamyl-transferase, alkaline phosphatase, creatinine, urea, and uric acid, and hence liver and renal dysfunction. LPS also increased serum levels of cholesterol and triglycerides and reduced glucose levels. Melatonin administration counteracted these organ and metabolic alterations at doses ranging between 20 and 60 mg/kg b. w. Melatonin significantly decreased lung lipid peroxidation and counteracted the LPS-induced NO levels in lungs and liver. Our results also show an inhibition of iNOS activity in rat lungs by melatonin in a dose-dependent manner. Expression of iNOS mRNA in lungs and liver was significantly decreased by melatonin (60 mg/kg b. w., 58-65%). We conclude that melatonin inhibits NO production mainly by inhibition of iNOS expression. The inhibition of NO levels may account for the protection of the indoleamine against LPS-induced endotoxemia in rats. PMID- 10463946 TI - Melatonin regulates glucocorticoid receptor: an answer to its antiapoptotic action in thymus. AB - We have previouslyreported that low doses of melatonin inhibit apoptosis in both dexamethasone-treated cultured thymocytes (standard model for the study of apoptosis) and the intact thymus. Here we elucidate the mechanism by which this agent protects thymocytes from cell death induced by glucocorticoids. Our results demonstrate an effect of melatonin on the mRNA for antioxidant enzymes in thymocytes, also showing an unexpected regulation by dexamethasone of these mRNA. Both an effect of melatonin on the general machinery of apoptosis and a possible regulation of the expression of the cell death related genes bcl-2 and p53 are shown not to be involved. We found melatonin to down-regulate the mRNA for the glucocorticoid receptor in thymocytes (glucocorticoids up-regulate their own receptor). The decrease by melatonin of mRNA levels for this receptor in IM-9 cells (where glucocorticoids down-regulate it) demonstrates that melatonin actually down-regulates glucocorticoid receptor. These findings allow us to propose the effects of melatonin on this receptor as the likely mediator of its thymocyte protection against dexamethasone-induced cell death. This effect of melatonin, given the oxidant properties of glucocorticoids, adds another mechanism to explain its antioxidant effects. PMID- 10463947 TI - Osmotic regulation of the heat shock response in H4IIE rat hepatoma cells. AB - The influence of cell hydration on the heat shock response was investigated in H4IIE hepatoma cells at the levels of HSP70 expression, MAP kinase activation, induction of c-jun and the MAP kinase phosphatase MKP-1, heat resistance, and development of tolerance/sensitization to arsenite after a priming heat treatment. Induction of HSP70, MKP-1, and c-jun by heat was delayed, but more pronounced or sustained, under hyperosmotic conditions compared with normo- and hypo-osmotically exposed cells. Anisosmolarity per se was ineffective to induce HSP70; some expression of the mRNAs for MKP-1 and c-jun in response to hyperosmolarity was found, but was small compared with the response to heat. Heat induced activation of JNK-1 was increased under hyperosmotic conditions and more sustained than the JNK-activity induced by hyperosmolarity at 37 degrees C. A prominent Erk-2 activation was found immediately after heat shock under hypo- and normo-osmotic conditions, but Erk-2 activation was weak in hyperosmolarity exposed cells. Despite anisosmotic alterations of the heat shock response at the molecular level, the heat resistance of H4IIE cells toward heat shock was not affected by ambient osmolarity. However, an osmolarity-dependent sensitization to arsenite was induced by a priming heat shock. The osmodependence of the H4IIE cell response to heat differs from that recently found in primary rat hepatocytes. The data are discussed in terms of cellular adaption mechanisms and their physiological relevance. PMID- 10463948 TI - Novel CXCR2-dependent liver regenerative qualities of ELR-containing CXC chemokines. AB - Severe acute liver injury due to accidental or intentional acetaminophen overdose presents a major clinical dilemma often requiring liver transplantation. In the present study, liver regeneration after profound liver injury in mice challenged with acetaminophen was facilitated by the exogenous addition of ELR-containing CXC chemokines such as macrophage inflammatory protein-2 (MIP-2), epithelial neutrophil-activating protein-78 (ENA-78), or interleukin 8. Intravenous administration of ELR-CXC chemokines or N-acetyl-cysteine (NAC) immediately after acetaminophen challenge in mice significantly reduced histological and biochemical markers of hepatic injury. However, when the intervention was delayed until 10 h after acetaminophen challenge, only ELR-CXC chemokines significantly reduced liver injury and mouse mortality. The delayed addition of ELR-CXC chemokines to cultured hepatocytes maintained the proliferation of these cells in a CXCR2-dependent fashion after acetaminophen challenge whereas delayed NAC treatment did not. These observations demonstrate that ELR-CXC chemokines represent novel hepatic regenerative factors that exhibit prolonged therapeutic effects after acetaminophen-induced hepatotoxicity. PMID- 10463949 TI - Death deflected: IL-15 inhibits TNF-alpha-mediated apoptosis in fibroblasts by TRAF2 recruitment to the IL-15Ralpha chain. AB - Interleukin-15 (IL-15) is a potent inhibitor of several apoptosis pathways. One prominent path toward apoptosis is the ligand-induced association of TNF receptor 1 (TNFR1) with death domain adaptor proteins. Studying if and how IL-15 blocks TNFR1-mediated apoptosis in a murine fibroblast cell line (L929), we show here that IL-15 blocks TNFR1-induced apoptosis via IL-15Ralpha chain signaling. The intracellular tail of IL-15Ralpha shows sequence homologies to the TRAF2 binding motifs of CD30 and CD40. Most important, binding of IL-15 to IL-15Ralpha successfully competes with the TNFR1 complex for TRAF2 binding, which may impede assembly of key adaptor proteins to the TNFR1 complex, and induces IkappaBalpha phosphorylation. Thus, IL-15Ralpha chain stimulation is a powerful deflector of cell death very early in the apoptosis signaling cascade, while TNF-alpha and IL 15 surface as major opponents in apoptosis control. PMID- 10463950 TI - Allergen mimotopes in food enhance type I allergic reactions in mice. AB - BIP1is a murine IgG antibody capable of enhancing the IgE binding to Bet v 1, the major birch pollen allergen. We have previously generated a mimotope of BIP1, designated Bet mim 1, from a constrained phage display peptide library. We demonstrated that oral immunization of BALB/c mice with the Bet mim 1 mimotope resulted in the induction of Bet v 1-specific IgG. The aim of this study was to test the influence of such an oral immunization with Bet mim 1 on a subsequent type I allergic response to Bet v 1. Phages displaying Bet mim 1 or control mimotopes, or PBS alone, were delivered to BALB/c mice by intragastric gavages prior to systemic sensitization with recombinant Bet v 1 and Al(OH)(3), an adjuvant inducing preferentially IgE antibody responses. Only mice fed with Bet mim 1-phages displayed substantially enhanced type I allergic skin reactivity to Bet v 1, as compared to mice pretreated with control mimotopes or PBS. A gastric digestion assay indicated that Bet v 1 and its homologue from apple, Mal d 1, were degraded within seconds under physiological conditions. In contrast, phage displayed mimotopes were resistant to digestion. Our data indicate that allergen mimics in the diet that resist digestion, can induce allergen specific IgG able to enhance an allergic response. We therefore conclude that sensitization via the oral route may represent a mechanism for aggravating type I allergic reactions, probably leading to an earlier onset of symptoms even at lower allergen dosage. PMID- 10463951 TI - Platelet-derived growth factor-induced activation of sphingosine kinase requires phosphorylation of the PDGF receptor tyrosine residue responsible for binding of PLCgamma. AB - Sphingosine-1-phosphate, a sphingolipid metabolite, is involved in the mitogenic response of platelet-derived growth factor (PDGF) and is formed by activation of sphingosine kinase. We examined the effect of PDGF on sphingosine kinase activation in TRMP cells expressing wild-type or various mutant betaPDGF receptors. Sphingosine kinase was stimulated by PDGF in cells expressing wild type receptors but not in cells expressing kinase-inactive receptors (R634). Cells expressing mutated PDGF receptors with phenylalanine substitutions at five major tyrosine phosphorylation sites 740/751/771/1009/1021 (F5 mutants), which are unable to associate with PLCgamma, phosphatidylinositol 3-kinase, Ras GTPase activating protein, or protein tyrosine phosphatase SHP-2, not only failed to increase DNA synthesis in response to PDGF but also did not activate sphingosine kinase. Moreover, mutation of tyrosine-1021 of the PDGF receptor to phenylalanine, which impairs its association with PLCgamma, abrogated PDGF induced activation of sphingosine kinase. In contrast, PDGF was still able to stimulate sphingosine kinase in cells expressing the PDGF receptor mutated at tyrosines 740/751 and 1009, responsible for binding of phosphatidylinositol 3 kinase and SHP-2, respectively. In agreement, PDGF did not stimulate sphingosine kinase activity in F5 receptor 'add-back' mutants in which association with the Ras GTPase-activating protein, phosphatidylinositol 3-kinase, or SHP-2 was individually restored. However, a mutant PDGF receptor that was able to bind PLCgamma (tyrosine-1021), but not other signaling proteins, restored sphingosine kinase sensitivity to PDGF. These data indicate that the tyrosine residue responsible for binding of PLCgamma is required for PDGF-induced activation of sphingosine kinase. Moreover, calcium mobilization downstream of PLCgamma, but not protein kinase C activation, appears to be required for stimulation of sphingosine kinase by PDGF.-Olivera, A., Edsall, J., Poulton, S., Kazlauskas, A., Spiegel, S. Platelet-derived growth factor-induced activation of sphingosine kinase requires phosphorylation of the PDGF receptor tyrosine residue responsible for binding of PLCgamma. PMID- 10463952 TI - Overexpression of manganese superoxide dismutase protects against mitochondrial initiated poly(ADP-ribose) polymerase-mediated cell death. AB - Mitochondria have recently been shown to serve a central role in programmed cell death. In addition, reactive oxygen species (ROS) have been implicated in cell death pathways upon treatment with a variety of agents; however, the specific cellular source of the ROS generation is unknown. We hypothesize that mitochondria-derived free radicals play a critical role in apoptotic cell death. To directly test this hypothesis, we treated murine fibrosarcoma cell lines, which expressed a range of mitochondrial manganese superoxide dismutase (MnSOD) activities, with respiratory chain inhibitors. Apoptosis was confirmed by DNA fragmentation analysis and electron microscopy. MnSOD overexpression specifically protected against cell death upon treatment with rotenone or antimycin. We examined bcl-x(L), p53 and poly(ADP-ribose) polymerase (PARP) to identify specific cellular pathways that might contribute to the mitochondrial-initiated ROS-mediated cell death. Cells overexpressing MnSOD contained less bcl-x(L) within the mitochondria compared to control (NEO) cells, therefore excluding the role of bcl-x(L). p53 was undetectable by Western analysis and examination of the proapoptotic protein bax, a p53 target gene, did not increase with treatment. Activation of caspase-3 (CPP-32) occurred in the NEO cells independent of cytochrome c release from the mitochondria. PARP, a target protein of CPP-32 activity, was cleaved to a 64 kDa fragment in the NEO cells prior to generation of nucleosomal fragments. Taken together, these findings suggest that mitochondrial-mediated ROS generation is a key event by which inhibition of respiration causes cell death, and identifies CPP-32 and the PARP-linked pathway as targets of mitochondrial-derived ROS-induced cell death. PMID- 10463953 TI - Neuronal survival activity of s100betabeta is enhanced by calcineurin inhibitors and requires activation of NF-kappaB. AB - S100betabeta is a calcium binding, neurotrophic protein produced by nonneuronal cells in the nervous system. The pathway by which it enhances neuronal survival is unknown. Here we show that S100betabeta enhances survival of embryonic chick forebrain neurons in a dose-dependent manner. In the presence of suboptimal amounts of S100betabeta, neuronal survival is enhanced by the immunosuppressants FK506 and cyclosporin A at concentrations that inhibit calcineurin, which is present in these cells. Rapamycin, an immunosuppressant that does not inhibit calcineurin, did not enhance cell survival. Cypermethrin, a direct and highly specific calcineurin inhibitor, mimicked the immunophilin ligands in its neurotrophic effect. None of the drugs stimulated neuronal survival in the absence of S100betabeta. In the presence of suboptimal amounts of S100betabeta, FK506, cyclosporin A, and cypermethrin (but not rapamycin) also increased NF kappaB activity, as measured by immunofluorescence of cells stained with antibody to the active subunit (p65) and by immunoblotting of nuclear extracts. Antioxidant and glucocorticoid inhibitors of NF-kappaB decreased both the amount of active NF-kappaB and the survival of neurons caused by S100betabeta alone or in the presence of augmenting drugs. We conclude that S100betabeta enhances the survival of chick embryo forebrain neurons through the activation of NF-kappaB. PMID- 10463954 TI - Regulation of neural differentiation by normal and mutant (G654A, amyloidogenic) gelsolin. AB - Gelsolin belongs to a family of proteins that modulate the structural dynamics of cytoskeletal actin. Gelsolin activity is required for the redistribution of actin occurring during membrane ruffling, cell crawling, and platelet activation. A point mutation (G654A) in the gelsolin gene causes a dominantly inherited systemic amyloidosis called familial amyloidosis of the Finnish type (FAF). This disease is characterized by a cranial neuropathy that cannot be explained solely by amyloid deposits. To address the question of whether gelsolin has a specific role in neural cell development, we transfected cDNA for wild type and G654A point-mutated gelsolin into a neural cell line, Paju, which can be induced to differentiate by treatment with phorbol 12-myristate 13-acetate. Overexpressed wild type gelsolin inhibited neural differentiation whereas mutated gelsolin did not, indicating that appropriate gelsolin activity is essential for neural sprouting. The G654A mutant gelsolin induced stabilization of F-actin and reduced the plasticity of neural development. This provides a novel etiopathogenetic mechanism for the neuronal dysfunction in FAF. PMID- 10463955 TI - Costimulatory action of glycoinositolphospholipids from Trypanosoma cruzi: increased interleukin 2 secretion and induction of nuclear translocation of the nuclear factor of activated T cells 1. AB - The effects of the glycoinositolphospholipids (GIPLs) fromTrypanosoma cruzi on T lymphocyte activation were investigated in a mouse T cell hybridoma (DO-11.10). Purified GIPLs from T. cruzi strains Y and G markedly increased IL-2 mRNA transcripts and IL-2 secretion induced by mitogenic anti-CD3 and anti-Thy1 mAbs. This costimulatory function was also revealed by the induction of IL-2 secretion after the simultaneous addition of the T. cruzi GIPLs and either the calcium ionophore A23187 or phorbol ester. The capacity of the GIPL molecule to induce an increase in cytoplasmic calcium levels was also demonstrated. After exposure of T cell hybridoma to GIPL, the nuclear transcription factor NFAT1 became partially dephosphorylated, and its nuclear localization was demonstrated both in the T cell hybridoma and in Balb/c CD3(+) cells. These results demonstrate that T. cruzi GIPL molecules are capable of signaling to T cells and therefore could be valuable tools for the study of T cell activation, besides playing a potential role in subverting the T lymphocyte immune response during T. cruzi infection. PMID- 10463957 TI - UK handwashing initiative. PMID- 10463958 TI - The great enemy lives on. PMID- 10463956 TI - Endogenous peroxynitrite mediates mitochondrial dysfunction in rat diaphragm during endotoxemia. AB - It has been shown that nitric oxide (NO), synthesized by the inducible NO synthase (iNOS) expressed in the diaphragm during endotoxemia, participates in the development of muscular contractile failure. The aim of the present study was to investigate whether this deleterious action of NO was related to its effects on cellular oxidative pathways. Rats were inoculated with E. coli lipopolysaccharide (LPS) or sterile saline solution (controls) and studied at 3 and 6 h after inoculation. iNOS protein and activity could be detected in the rat diaphragm as early as 3 h after LPS, with a sustained steady-state concentration of 0.5 microM NO in the muscle associated with increased detection of hydrogen peroxide (H(2)O(2)). In vitro, the same NO concentration produced a marked increase in H(2)O(2) production by isolated control diaphragm mitochondria, thus reflecting a higher intramitochondrial concentration of nondiffusible superoxide anion (O(2)(-.)). In a similar way, whole diaphragmatic muscle and diaphragm mitochondria from endotoxemic rats showed a progressive increase in H(2)O(2) production associated with uncoupling and decreased phosphorylating capacity. Simultaneous with the maximal impairment in respiration (6 h after LPS), nitration of mitochondrial proteins (a peroxynitrite footprint) was detected and diaphragmatic force was reduced. Functional mitochondrial abnormalities, nitration of mitochondrial proteins, and the decrease in force were significantly attenuated by administration of the NOS inhibitor L-NMMA. These results show that increased and sustained NO levels lead to a consecutive formation of O(2)(-.) that reacts with NO to form peroxynitrite, which in turn impairs mitochondrial function, which probably contributes to the impairment of muscle contractility. during endotoxemia. PMID- 10463959 TI - Pulmonary hemorrhage in factor V deficiency. PMID- 10463961 TI - Cost effectiveness of transbronchial needle aspiration. AB - OBJECTIVE: To evaluate the yield and cost effectiveness of transbronchial needle aspiration (TBNA) in the assessment of mediastinal and/or hilar lymphadenopathy. DESIGN: Retrospective study. SETTING: A university hospital. POPULATION STUDIED: Ninety-six patients referred for bronchoscopy with computed tomographic evidence of significant mediastinal or hilar adenopathy. RESULTS: Ninety-nine patient records were reviewed. Three patients had two separate bronchoscopy procedures. TBNA was positive in 42 patients (44%) and negative in 54 patients. Of the 42 patients with a positive aspirate, 40 had malignant cytology and two had cells consistent with benign disease. The positive TBNA result altered management in 22 of 40 patients with malignant disease and one of two patients with benign disease, thereby avoiding further diagnostic procedures. The cost of these subsequent procedures was estimated at $27,335. No complications related to TBNA were documented. CONCLUSIONS: TBNA is a high-yield, safe and cost effective procedure for the diagnosis and staging of bronchogenic cancer. PMID- 10463960 TI - Nonasthmatic chronic cough: No effect of treatment with an inhaled corticosteroid in patients without sputum eosinophilia. AB - BACKGROUND: Inhaled corticosteroids are effective in suppressing a chronic cough without asthma associated with sputum eosinophilia. OBJECTIVE: To investigate the inflammatory characteristics in the induced sputum of patients with a chronic cough without asthma or known cause and the effects of budesonide treatment on chronic cough in those patients. PATIENTS AND METHODS: Forty-four adults (mean [minimu, maximum] age of 45 years [20,75], 28 women, 17 atopic subjects and 32 nonsmokers], with a daily bothersome cough for at least one year and who had no evidence of asthma or other known cause for the cough, were consecutively enrolled. The trial was a randomized, double-blind, controlled parallel group trial of budesonide 400 mg twice daily for two weeks versus placebo. Patients then received open administration of the same dose of budesonide for a further two weeks. Sputum was induced before and at the end of each treatment period. Cough severity was documented by a visual analogue scale. RESULTS: Thirty-nine (89%) patients produced mucoid sputum after induction on at least one study visit. At baseline, the majority (59%) had a mild elevation in the median proportion of neutrophils (65%). All had elevated fluid phase levels of fibrinogen (3200 mg/L) and albumin (880 mg/L), and high levels of interleukin-8 and substance P. Interleukin-8 correlated with neutrophils (rho=0.72, P<0.001), fibrinogen (rho=0.65, P<0.001), albumin (rho=0.67, P=0. 001) and eosinophil cationic protein (rho=0.60, P=0.001). Substance P correlated with albumin (rho=0.60, P=0.006). No subject had an increase in eosinophils. Treatment with budesonide did not affect cough or sputum measurements. CONCLUSIONS: Patients with nonasthmatic chronic cough enrolled in this study had evidence of a mild neutrophilia and/or microvascular leakage. Chronic cough did not respond to treatment with budesonide, perhaps because the cause was not associated with sputum eosinophilia. PMID- 10463962 TI - Management of exercise-induced bronchoconstriction. AB - Exercise-induced bronchoconstriction (EIB) is a common clinical manifestation of asthma, occurring in 70% to 80% of asthmatics. Evidence suggests that exercise and the ensuing bronchoconstriction do not contribute to a worsening of asthmatic inflammation. Asthmatics should not be discouraged from exercising, and, with adequate management, most patients should be able to exercise regularly with only minor symptoms. The first step in the management of patients with EIB should be to obtain optimum control of the underlying asthma, often requiring regular treatment with inhaled steroids. Regular treatment with inhaled corticosteroids usually reduces the extent of EIB by 50% or more. Frequently, despite optimal management of the underlying asthma, patients develop EIB symptoms requiring additional treatment. Short and long acting inhaled beta2-agonists are highly effective at reducing the magnitude of EIB, although there are concerns that the extent of protection diminishes during periods of regular use of these agents. Inhaled cromolyn and nedocromyl are effective at reducing the extent of EIB in some patients, although this protection does not extend beyond 2 to 3 h after treatment. The recently developed leukotriene receptor antagonists are effective at reducing the extent of EIB by 40% to 70%, and have the advantage that this protection lasts throughout the day and does not appear to diminish with regular use. Other agents, including anticholinergics and antihistamines, have been shown to offer partial protection against EIB, suggesting the possibility of using a combination treatment to manage some patients' symptoms. Finally, there is encouraging evidence suggesting that modifications of the pattern of exercise can markedly reduce the extent of EIB. PMID- 10463963 TI - Delayed diaphragmatic herniation masquerading as a complicated parapneumonic effusion. AB - Injury to the diaphragm following blunt or penetrating thoracoabdominal trauma is not uncommon. Recognition of this important complication of trauma continues to be a challenge because of the lack of specific clinical and plain radiographic features, the frequent presence of other serious injuries and the potential for delayed presentation. Delayed diaphragmatic herniation often presents with catastrophic bowel obstruction or strangulation. Early recognition of diaphragmatic injury is required to avoid this potentially lethal complication. The case of a 35-year-old man with a history of a knife wound to the left flank 15 years previously, who presented with unexplained acute hypoxemic respiratory failure and a unilateral exudative pleural effusion that was refractory to tube thoracostomy drainage, is reported. After admission to hospital, he developed gross dilation of his colon; emergency laparotomy revealed an incarcerated colonic herniation into the left hemithorax. Interesting clinical features of this patient's case included the patient's hobby of weightlifting, a persistently deviated mediastinum despite drainage of the pleural effusion and deceptive pleural fluid biochemical indices. PMID- 10463964 TI - What is the significance of race to prostate carcinoma? PMID- 10463965 TI - Giving bad news. Is there a kinder, gentler way? PMID- 10463966 TI - Hepatocellular carcinoma and coinfection with hepatitis B and C. Making a difficult situation worse. PMID- 10463967 TI - The genetic epidemiology of male breast carcinoma. PMID- 10463968 TI - Laparoscopy. To inflate or lift? PMID- 10463969 TI - Laparoscopy. To inflate or lift? PMID- 10463970 TI - Diet and breast carcinoma survival. An abundance of hope, a dearth of evidence. PMID- 10463971 TI - Diet and breast carcinoma survival: An abundance of hope, a dearth of evidence: reply. PMID- 10463972 TI - Expression of the cyclin-dependent kinase inhibitor p21(WAF1/CIP1) and p53 tumor suppressor in dysplastic progression and adenocarcinoma in Barrett esophagus. AB - BACKGROUND: Barrett esophagus predisposes individuals to esophageal carcinoma, which develops from intermediate stages of tissue dysplasia primarily in the vicinity of the gastroesophageal junction. Understanding the cellular and molecular events in the progression of Barrett esophagus to adenocarcinoma may contribute to its early diagnosis and treatment. Mutation and overexpression of the tumor suppressor p53 have previously been observed in Barrett high grade dysplasia and adenocarcinoma. The expression of the cyclin-dependent kinase (CdK) inhibitor p21 can be up-regulated by p53, resulting in the down-regulation of cell division at the G(1)/S-phase transition. The current study examined the correlation between the expression of p21 and p53 by quantifying their levels during the progression of dysplasia and adenocarcinoma in Barrett esophageal tissues. METHODS: Barrett esophageal tissue samples that were negative or indefinite for dysplasia, contained dysplasia, and contained adenocarcinoma were examined by immunohistochemistry. Paraffin embedded sections of lining and glandular epithelia were adsorbed with primary murine antibodies against human p21 or p53 followed by horseradish peroxidase secondary antibody. An immunoreactivity score for each primary antibody and section was obtained by multiplying a staining intensity factor by the percent of positively stained cells. RESULTS: Nuclear p21 expression was detectable immunohistochemically in Barrett esophagus that was negative for dysplasia, but it was significantly elevated (P 50,000 ng/mL and tumor volume >30% were significant predictive variables associated with the presence of paraneoplastic syndromes in HCC patients. CONCLUSIONS: HCC patients with paraneoplastic syndromes usually had higher levels of serum AFP and larger tumor volumes than those without. Patients with HBV-related HCC had a significantly higher prevalence of paraneoplastic syndromes than those with HCV-related HCC. PMID- 10463979 TI - Multidrug resistance protein expression in chronic myeloid leukemia: associations and significance. AB - BACKGROUND: Overexpression of the multidrug resistance gene (MDR1) product, the MDR1 protein (MDR1), has been associated with poor prognosis in several hematologic malignancies. The significance of MDR1 levels in patients with chronic myeloid leukemia (CML) has not been established. METHODS: The authors investigated MDR1 levels and their association with patient and tumor characteristics, responsiveness to therapy, and long term prognosis in 198 CML patients. These included 127 patients in early chronic phase (ECP) CML, 31 patients in late chronic phase (LCP) CML, and 40 patients in accelerated or blastic phase CML. MDR1 expression was analyzed by Western blot analysis and quantitative solid-phase plate radioimmunoassay. MDR1 levels were measured on cell lysates obtained from the bone marrow mononuclear cell fraction. Expression was compared in relation to the median derived from 36 normal control samples. RESULTS: Among patients with CML, high levels of MDR1 were found in 73 of 127 ECP (57%), 20 of 31 LCP (65%), 8 of 27 in accelerated phase (30%), and 8 of 13 in blastic phase (62%) (P value not significant). Furthermore, among the 127 ECP CML patients, high MDR1 levels were associated with age >/=50 years (69% vs. 51%; P < 0. 05), thrombocytosis >700 x 10(9)/L (84% vs. 53%; P < 0.01), and leukocyte counts 299 mg/day of oral morphine equivalent), what the characteristics of these patients are, and whether the use of HDM might affect their survival. METHODS: The authors retrospectively studied the medical records of all 651 inpatients hospitalized at their center between January 1996 and December 1997. Information was collected regarding demographic parameters, medical diagnosis, pain mechanism, morphine dosage, use of rescue doses in addition to regular doses, use of coanalgesics and adjuvant treatments, and survival time in the hospice setting as associated with morphine dosage. RESULTS: The authors identified 453 patients (69.58%) who received morphine for pain relief, of whom 55 (12.14%) needed more than 299 mg/day. Morphine dosage was negatively associated with age (r = -0.35, P = 0.01). Male patients and nonwhite patients required slightly higher dosages than others. Primary breast and genitourinary cancers, as well as metastases to bone and spinal diseases, were associated with higher morphine dosages. Statistical analysis indicated a positive correlation between the log maximum of morphine dosage and the total number of rescue dosages (r = 0.307, P = 0.025). The median survival of patients on HDM was 15.6 days and did not differ from the survival of patients taking a lower dosage. CONCLUSIONS: A fairly strong correlation exists between morphine dosage and some clinicodemographic data. No significant dose-limiting adverse effects were observed, suggesting a high clinical safety profile. High morphine dosage does not affect patient survival. Awareness of the dosage factors will improve our ability to treat and predict probable HDM dosage, thus shortening the period until pain relief is reached. PMID- 10463989 TI - Leptomeningeal melanoma in childhood. AB - BACKGROUND: Malignant melanoma (MM) is one of the least common types of childhood cancer, accounting for less than 1% of all pediatric malignancies. Neurocutaneous melanosis (NCM) is a rare phakomatosis consisting of congenital abnormal pigmentation of the skin and meninges. The meningeal lesions are particularly prone to malignant change. METHODS: The authors describe 5 patients with NCM and 1 with primary leptomeningeal melanoma (LMM) seen at 2 treatment centers in the north of England over a 13-year period (1984-1997). RESULTS: The clinical features, progress, radiological findings, and treatment of these patients are discussed. All six died within eight months of their diagnosis, illustrating the difficulties faced in treating patients with these conditions. The authors reviewed the published literature on NCM, concentrating on the various therapeutic strategies that have been tried. Very little consistency in approach was found. Malignant skin lesions in NCM may be less responsive than primary malignant melanoma, but the small number of patients with primary LMM or brain metastases of MM make comparisons with NCM difficult. The authors' own series illustrates well the piecemeal nature of therapy for patients with these rare conditions. CONCLUSIONS: The rate of incidence of MM melanoma in the U.K. is increasing, and it will represent an increasing proportion of the pediatric oncologist's workload. A consistent approach to the therapy of patients with metastatic MM and NCM is needed if we are to have any hope of offering more than palliative therapy to these children in the future. PMID- 10463990 TI - Communication skills training in oncology. Description and preliminary outcomes of workshops on breaking bad news and managing patient reactions to illness. AB - BACKGROUND: Cancer clinicians do not receive routine training in the psychosocial aspects of patient care such as how to communicate bad news or respond to patients who have unrealistic expectations of cure. Postgraduate workshops may be an effective way to increase interpersonal skills in managing these stressful patient encounters. METHODS: The authors conducted 2 half-day workshops for oncology faculty, one on breaking bad news and one on dealing with "problem situations." Participants met in a large group for didactic presentations and then small groups in which they used role-play and discussion to problem-solve difficult cases from their practices. The small groups were assisted in their work by trained physician facilitators. The workshops were evaluated by means of a follow-up satisfaction questionnaire as well as a self-efficacy measure, which was administered before and after the workshops. RESULTS: Twenty-seven faculty and 2 oncology fellows participated in the training programs. Satisfaction questionnaires showed that the programs met the educational objectives and were considered to be useful and relevant by the participants. Self-efficacy questionnaires revealed an increase in confidence in communicating bad news and managing problem situation cases from before to after the workshop. The majority of attendees welcomed the opportunity to discuss their difficult cases with colleagues. A number resolved to implement newly learned approaches to common patient problems they encountered frequently. CONCLUSIONS: Communication skills workshops may be a useful modality to provide training to oncologists in stressful aspects of the physician-patient relationship. Further research is needed to assess whether long term benefits accrue to the participants. PMID- 10463991 TI - Human tumor xenografts in nude mice are not always of human origin. A warning signal. PMID- 10463992 TI - Author reply PMID- 10463994 TI - Author reply PMID- 10463993 TI - Front-line chemotherapy with cisplatin and etoposide for patients with brain metastases from breast carcinoma, nonsmall cell lung carcinoma, or malignant melanoma. A prospective study. PMID- 10463995 TI - 24-hour ECG frequency-domain measures in preeclamptic and healthy pregnant women during and after pregnancy. AB - OBJECTIVE: The aim of the present study was to evaluate the autonomic balance in women with preeclampsia and in healthy women during and after pregnancy by means of a 24-h ECG Holter recording combined with power spectral analysis. METHODS: Fifteen preeclamptic and 15 healthy women underwent 24-h Holter monitoring in the 32nd-36th week of gestation and 3-6 months postpartum. The power spectrum of the maternal electrocardiogram was analyzed with an autoregressive algorithm. MAIN OUTCOME MEASURES: The power spectrum contains two major components: a low frequency peak, primarily attributed to sympathetic tone, and a high-frequency peak, reflecting vagal tone. RESULTS: The power spectrum of maternal heart rate variability did not differ between preeclamptic and normal women during pregnancy. After delivery, the amplitude of all components became significantly higher than those during pregnancy, with one exception: the high-frequency component in the patients who had been preeclamptic. In a comparison of the two groups, the high-frequency component after delivery was significantly lower in women who had preeclampsia than in normal healthy women (p = 0.03). CONCLUSIONS: During pregnancy, the power spectrum is reduced and cannot be used to distinguish between patients with preeclampsia and normal healthy women. Three to 6 months after delivery, the high-frequency component is still reduced in the preeclamptic group of women. This indicates an impaired vagal modulation even in the nonpregnant state in this group of women, unlike those who had a normotensive pregnancy. PMID- 10463996 TI - Evidence of a nonendothelial source of nitric oxide in the isolated perfused hindlimb vasculature of the pregnant rat. AB - OBJECTIVE: This study was undertaken to further elucidate the roles of the vascular endothelium and nitric oxide (NO) in the regulation of vascular tone and constrictor responsiveness in pregnancy. METHODS: The perfusion pressure-flow relationship was measured in isolated, perfused, norepinephrine-constricted (1) endothelium-intact, (2) endothelium-denuded, and (3) N omega-nitro-L-arginine methyl ester (L-NAME)-treated hindlimbs from nonpregnant and term-pregnant rates. RESULTS: Baseline perfusion pressure at a flow rate of 2 mL/min was similar (approximately 20 min Hg) in all hindlimbs. Norepinephrine (0.5 muM) increased perfusion pressure in both nonpregnant (+21.6 +/- 2.4 mm Hg) and pregnant (+13.6 +/- 0.9 mm Hg) endothelium-intact rat hindlimbs. In nonpregnant rat hindlimbs, endothelium removal and L-NAME increased norepinephrine vasoconstriction similarly (+44.3 +/- 4.0 mm Hg and +46.4 +/- 8.6 mm Hg, respectively). In pregnant-rat hindlimbs, L-NAME increased norepinephrine vasoconstriction by 43.5 +/- 10.8 mm Hg, similar to that in nonpregnant-rat hindlimbs, but endothelium removal only increased norepinephrine vasoconstriction by 28.0 +/- 2.2 mm Hg. Perfusion pressure increased linearly as the flow rate was increased from 2 to 4 mL/min, and the slope of the regression line of the endothelium-intact pregnant rat hindlimbs (7.0 +/- 0.6) was slightly, but not significantly, lower than that of the nonpregnant-rat hindlimbs (9.6 +/- 0.9). Endothelium removal increased the slopes of the regression lines, but that of pregnant-rat hindlimbs (12.8 +/- 1.6) was significantly lower (p < or = 0.05) than that of the nonpregnant-rat hindlimbs (23.8 +/- 1.8). L-NAME caused a similar increase in the pressure-flow slopes of nonpregnant-rat (36.5 +/- 3.4) and pregnant-rat (32.1 +/- 5.3) hindlimbs. CONCLUSIONS: These results suggest that nonendothelial nitric oxide production may be increased in the hindlimb resistance vasculature of the pregnant rat, which may play a role in the normal pregnancy blunting of constrictor responsiveness and reduction of vascular resistance. PMID- 10463997 TI - Maternal factors modulate the increase in vasoactive substances during rat pregnancy. AB - OBJECTIVE: To explore if the changes in vasoactive substances observed during early pregnancy in the rat are modulated by maternal or fetoplacental factors. METHODS: Urinary excretion of cGMP, 6-keto-prostaglandin-F1 alpha (6-keto-PGF1 alpha), thromboxane B2 and kallikrein activity was measured in pregnant (P, n = 11), pseudopregnant (PSP, n = 12), and virgin (n = 13) rats and in ovariectomized virgin rats supplemented with slow-release pellets containing either progesterone (50 mg/pellet) or estradiol (0.5 mg/pellet) or a combination of both hormones, for 21 days. RESULTS: The cGMP excretion was higher in PSP rats than in virgin rats at day 5 (virgin = 82 +/- 7, P = 93 +/- 5, PSP = 110 +/- 8 nmol/24 h, p < 0.05); at day 10, values were significantly increased in P and PSP rats. 6-keto PGF1 alpha excretion was similarly elevated in P and PSP rats at day 5 (virgin = 120 +/- 10, P = 160 +/- 10, and PSP = 174 +/- 14 ng/24 h, p < 0.01). This trend was still present at day 10. Thromboxane B2 excretion showed a nonsignificant increase in P and PSP rats in day 5; at day 10, values were significantly elevated in both experimental groups (virgin = 23 +/- 2, P = 32 +/- 4, and PSP = 32 +/- 2 ng/24 h, p < 0.05). Kallikrein excretion was significantly increased in PSP and P rats at days 5 and 10. Estradiol or progesterone administration caused a significant decrease in serum aldosterone and an increase in urinary kallikrein activity. CONCLUSIONS: These results indicate that during the first half of rat pregnancy, the increment in vasoactive substances is modulated by maternal and not by fetoplacental factors. PMID- 10463998 TI - Platelet sodium pump and sodium potassium cotransport activity in nonpregnant, normotensive, and hypertensive pregnant women. AB - OBJECTIVE: To determine ouabain-sensitive sodium pump and bumetanide-sensitive sodium potassium cotransport activity in platelets from nonpregnant and normotensive pregnant women and from women with pregnancy-induced hypertension (PIH). METHODS: Blood was collected from 9 normotensive nonpregnant subjects, 24 normotensive pregnant subjects in both second and third trimesters, 9 subjects who developed proteinuric PIH, and 9 subjects who developed moderate nonproteinuric PIH. Platelet sodium pump activity was determined by the difference in the uptake of rubidium-86 in the presence and absence of ouabain; sodium potassium cotransport (SPC) activity is that component that is inhibitable by bumetanide. RESULTS: SPC activity was similar in normotensive subjects in the second [median (range) 78 mmol Rb/h/mg protein (18-140)] and third trimesters [85 (39-134)] but was significantly (p < 0.001) higher than in nonpregnant subjects [22 (4-107)]. In addition, SPC was significantly (p < 0.001) lower in subjects with nonproteinuric [42 (4-67)] or proteinuric PIH [59 (33-102)] compared to those who remained normotensive. Sodium pump activity was significantly higher (p < 0.05) in nonpregnant subjects [263 (188-430)] compared with the other groups of subjects. Total rubidium uptake was significantly higher (p < 0.05) in third trimester normotensive subjects [471 (243-560)] compared with second-trimester subjects [405 (278-608)]. CONCLUSION: Our results suggest that the lower SPC activity in both nonproteinuric and proteinuric PIH may be an early sign of abnormality in the transport of sodium and potassium across the vascular smooth muscle cell membrane, which is responsible for the maintenance of blood pressure. PMID- 10463999 TI - Serum levels of ELAM-1, but not CD44, predict the clinical outcome of patients with preeclampsia. AB - OBJECTIVE: Preeclampsia is a severe complication in pregnancy, causing considerable maternal and fetal morbidity and mortality. Experimental evidence indicates that adhesion molecules are key factors of endothelial activation in preeclampsia. The aim of our study was to evaluate if serum levels of adhesion molecules CD44 and ELAM-1 provide clinically useful information as prognostic markers for preeclampsia. METHODS: A matched-pair study including 43 women with preeclampsia and 43 women with uncomplicated pregnancies was performed. Enzyme linked immunosorbent assay (ELISA) was used to determine serum levels of CD44 and ELAM-1. Results were correlated with clinical data. RESULTS: Median serum levels of ELAM-1 in controls and in women with preeclampsia were 8.9 ng/mL (minimum 0, maximum 20.0) and 12.0 ng/mL (minimum 4.0, maximum 27.0), respectively (Kruskal Wallis test, p = 0.01). In a univariate logistic regression model, ELAM-1 did reveal a significant influence on the odds of presenting with preeclampsia as well as on the odds of premature termination of the pregnancy due to preeclampsia (univariate logistic regression, p = 0.03 and p = 0.01, respectively). The risk of premature termination of the pregnancy was 0.5%, 15.3%, and 80.5% at ELAM-1 serum levels of 0 ng/mL, 10 ng/mL, and 20 ng/mL, respectively. No significant correlation between CD44 serum levels and clinicopathological parameters due to preeclampsia was observed. CONCLUSIONS: If these results are confirmed in a larger series, ELAM-1 could be used as a prognostic factor in preeclamptic women, allowing early identification and appropriate management of high-risk patients with preeclampsia. It is unlikely that measurement of ELAM-1 will be of value as a screening test. PMID- 10464000 TI - Mechanisms of endothelium-dependent relaxation in myometrial resistance vessels and their alteration in preeclampsia. AB - OBJECTIVE: To investigate the importance of prostacyclin and nitric oxide synthesis in endothelium-dependent relaxation in myometrial resistance vessels, and to test the hypothesis that a deficiency in nitric oxide synthesis contributes to the known alterations in endothelial function in preeclampsia. METHODS: Thirty-six women with normal pregnancies and 14 with preeclampsia had the myometrium biopsied at cesarean section. Resistance arteries were dissected and mounted on a wire myograph. After preconstriction with vasopressin, vessels were treated cumulatively with bradykinin. The process was repeated in the presence of indomethacin and then indomethacin and NG-monomethyl-L-arginine (L NMMA). RESULTS: The vessels from women with normal pregnancies showed endothelium dependent relaxation to bradykinin which was not significantly altered by the presence of indomethacin. The addition of L-NMMA significantly, but only partially, reduced the relaxation to bradykinin in the presence of indomethacin (p = 0.03). The vessels from women with preeclampsia showed markedly reduced relaxation to bradykinin (p < 0.0001), as compared to vessels from normal pregnant women. Relaxation of vessels from women with preeclampsia was increased by the addition of indomethacin (p = 0.03) but was virtually eradicated by the presence of L-NMMA. CONCLUSIONS: Eicosanoid synthesis plays little part in the relaxation of normal pregnant myometrial vessels to bradykinin. Nitric oxide synthesis mediates part but not all of the endothelium-dependent relaxation. In preeclampsia, relaxation to bradykinin is reduced; inhibition of eicosanoid synthesis allows increased relaxation, and nitric oxide synthesis appears to mediate a greater proportion of the relaxation than in normal pregnant women. PMID- 10464001 TI - Are women with polycystic ovary syndrome at an increased risk of pregnancy induced hypertension and/or preeclampsia? AB - OBJECTIVE: To study whether there is an increased risk of glucose intolerance and hypertensive complications during pregnancy in women with polycystic ovary syndrome (PCOS) and if there is an adverse pregnancy outcome. METHODS: In a retrospective case-control study, pregnancies and neonatal outcome were compared in 33 women with PCOS and 66 women without PCOS. The women were treated at Huddinge University Hospital; antenatal care was given at associated outpatient units. MAIN OUTCOME MEASURES: Blood glucose and blood pressure during the different trimesters. Pregnancy outcome in terms of gestational length, birth weight, and need for neonatal intensive care. RESULTS: No significant differences were found in blood glucose levels between the groups. There were also no differences in blood pressure during the first and second trimester. However, during the third trimester and labor, the PCOS group had a significantly higher blood pressure than the control group. Apart from a tendency toward reduced growth of twins in PCOS mothers, the babies were healthy, overall, with few problems in the neonatal period. CONCLUSIONS: No major differences with regard to perinatal outcome in pregnant women with and without PCOS were found. An increased risk of hypertensive disorders in the third trimester and during labor was demonstrated in the PCOS group. This suggests that in the antenatal care of women with PCOS, attention should focus on blood pressure in order to reduce the risks of morbidity associated with hypertension. PMID- 10464002 TI - The eNOS gene: a candidate for the preeclampsia susceptibility locus? AB - OBJECTIVE: To investigate the endothelial cell nitric oxide synthase (eNOS) gene as a candidate for susceptibility to preeclampsia. METHODS: Twenty-six Australian families containing 11 eclamptics, 59 severe preeclamptics, and 27 mild preeclamptics were used to test for linkage between the eNOS gene region and preeclampsia. Two microsatellite markers (D7S483 and D7S505) in the proximity of the eNOS gene were used. MAIN OUTCOME MEASURES: Logarithm of odds (LOD) scores were used to examine the cosegregation of alleles with the disease under a variety of inheritance models. Model-independent analysis, affected pedigree member method (AFFPED), and pairwise haplotype sharing between affected sibs were also used. RESULTS: Two-point LOD score analysis gave no evidence of linkage between preeclampsia and two markers in close proximity to the eNOS gene (LOD scores < 1) for any of the inheritance models investigated, with no evidence of heterogeneity between pedigrees. The AFFPED and the pairwise haplotype sharing test on affected sibs also gave no evidence of linkage (p-values > 0.05). CONCLUSION: This study provides no evidence for linkage between two markers in close proximity to the eNOS gene and preeclampsia in these families. These results do not support the recent suggestion that eNOS could be a familial pregnancy-induced hypertension gene (Arngrimsson R, et al., Am J Hum Genet 1997;61:354-62). Distinguishing preeclampsia from other hypertensive disorders in pregnancy is difficult. Hypertension appears to be a consequence, rather than a primary cause of preeclampsia. Given the vasodilatory role of the eNOS gene product, it is possible that the linkage recently reported for eNOS reflects its relationship with hypertension rather than preeclampsia. PMID- 10464003 TI - Concentrations of serum granulocyte-colony-stimulating factor in normal pregnancy and preeclampsia. AB - OBJECTIVE: In order to elucidate the potential role of granulocyte-colony stimulating factor (G-CSF) during the course of normal pregnancy and preeclampsia, we measured the serum concentrations of G-CSF in both normal pregnant women and preeclamptic patients. METHODS: Sera of 10 nonpregnant women, 34 normal pregnant women (n = 10, first trimester; n = 10, second trimester; n = 14, third trimester), 10 postpartum women, 10 mild preeclamptic patients, and 10 severe preeclamptic patients were collected. The serum concentrations of G-CSF were measured by enzyme immunoassay. RESULTS: The serum level of G-CSF in normal pregnant women (third trimester: 38.3 +/- 15.3 pg/mL; mean +/- SD) was significantly increased when compared with the levels observed in nonpregnant women (20.3 +/- 10.1 pg/mL, p < 0.05), which was similar to the G-CSF concentrations in postpartum subjects (20.7 +/- 9.1 pg/mL). The mild and severe preeclamptic patients showed significantly higher levels of G-CSF (56.9 +/- 18.8 pg/mL, p < 0.05; 73.2 +/- 30.4 pg/mL, p < 0.0001, respectively) than those noted in the third trimester women. The preeclamptic patients who presented with edema had greater (p < 0.01) serum levels of G-CSF (75.5 +/- 25.0 pg/mL) compared with nonedematous patients (44.7 +/- 14.9 pg/mL). The serum levels of G-CSF significantly correlated with both weight gain (p < 0.05), diastolic blood pressure (p < 0.01), and systolic blood pressure (p < 0.01), but not with white blood cell counts. CONCLUSION: Serum concentrations of G-CSF are increased in normal and, even more so, in preeclamptic pregnancies. Because there was no relationship between serum G-CSF concentration and the number of leukocytes, G CSF might act not to promote the physiological leukocytosis of pregnancy, but to stimulate the function of leukocytes such as phagocytosis. Moreover, it might be that G-CSF plays important roles in the activation of granulocytes or vascular endothelial injury, which are considered to be important pathological conditions in the development of preeclampsia. PMID- 10464004 TI - 24-hour ambulatory blood pressure monitoring: a comparison between transdermal glyceryl-trinitrate and oral nifedipine. AB - The objective of the present study is to compare the effectiveness of transdermal glyceryl-trinitrate versus oral nifedipine in lowering blood pressure in patients affected by pregnancy-induced hypertension (PIH). Thirty-six consecutive pregnant women have been evaluated at different gestational ages after the diagnosis of PIH or preeclampsia (PE). After a 24-h ambulatory blood pressure monitoring, patients were allocated to three groups: those receiving oral nifedipine and those receiving transdermal glyceryl-trinitrate in a continuous (24 h/day) or intermittent (16 h/day) administration. A second blood pressure monitoring was performed after 2 weeks of treatment. Systolic and diastolic blood pressure were compared by using the Cosinor method looking at mesor, amplitude, and acrophase. Baseline systolic and diastolic blood pressure was similar among the three groups. Neither the transdermal glyceryl-trinitrate administered for 24 or 16 h nor oral nifedipine affected systolic and diastolic blood pressure. Analysis of variance showed that the posttreatment values were similar among the groups. Further studies are needed to verify the possible use of transdermal glyceryl trinitrate as an antihypertensive drug during pregnancy. PMID- 10464005 TI - Discovery of novel disaccharide antibacterial agents using a combinatorial library approach. PMID- 10464006 TI - PTP1B inhibition and antihyperglycemic activity in the ob/ob mouse model of novel 11-arylbenzo[b]naphtho[2,3-d]furans and 11-arylbenzo[b]naphtho[2,3-d]thiophenes. PMID- 10464007 TI - Structure-based identification of small molecule antiviral compounds targeted to the gp41 core structure of the human immunodeficiency virus type 1. AB - Recent X-ray crystallographic determination of the HIV-1 envelope glycoprotein gp41 core structure opened up a new avenue to discover antiviral agents for chemotherapy of HIV-1 infection and AIDS. We have undertaken a systematic study to search for anti-HIV-1 lead compounds targeted to gp41. Using molecular docking techniques to screen a database of 20 000 organic molecules, we found 16 compounds with the best fit for docking into the hydrophobic cavity within the gp41 core and with maximum possible interactions with the target site. Further testing of these compounds by an enzyme-linked immunosorbent assay and virus inhibition assays discerned two compounds (ADS-J1 and ADS-J2) having inhibitory activity at micromolar concentrations on the formation of the gp41 core structure and on HIV-1 infection. These two compounds will be used as leads to design more effective HIV-1 inhibitors targeted to the HIV-1 gp41 core structure. PMID- 10464008 TI - Lead generation using pharmacophore mapping and three-dimensional database searching: application to muscarinic M(3) receptor antagonists. AB - By using a pharmacophore model, a geometrical representation of the features necessary for molecules to show a particular biological activity, it is possible to search databases containing the 3D structures of molecules and identify novel compounds which may possess this activity. We describe our experiences of establishing a working 3D database system and its use in rational drug design. By using muscarinic M(3) receptor antagonists as an example, we show that it is possible to identify potent novel lead compounds using this approach. Pharmacophore generation based on the structures of known M(3) receptor antagonists, 3D database searching, and medium-throughput screening were used to identify candidate compounds. Three compounds were chosen to define the pharmacophore: a lung-selective M(3) antagonist patented by Pfizer and two Astra compounds which show affinity at the M(3) receptor. From these, a pharmacophore model was generated, using the program DISCO, and this was used subsequently to search a UNITY 3D database of proprietary compounds; 172 compounds were found to fit the pharmacophore. These compounds were then screened, and 1-[2-(2 (diethylamino)ethoxy)phenyl]-2-phenylethanone (pA(2) 6.67) was identified as the best hit, with N-[2-(piperidin-1-ylmethyl)cycohexyl]-2-propoxybenz amide (pA(2) 4. 83) and phenylcarbamic acid 2-(morpholin-4-ylmethyl)cyclohexyl ester (pA(2) 5.54) demonstrating lower activity. As well as its potency, 1-[2-(2 (diethylamino)ethoxy)phenyl]-2-phenylethanone is a simple structure with limited similarity to existing M(3) receptor antagonists. PMID- 10464009 TI - Quantitative structure-activity relationship modeling of dopamine D(1) antagonists using comparative molecular field analysis, genetic algorithms partial least-squares, and K nearest neighbor methods. AB - Several quantitative structure-activity relationship (QSAR) methods were applied to 29 chemically diverse D(1) dopamine antagonists. In addition to conventional 3D comparative molecular field analysis (CoMFA), cross-validated R(2) guided region selection (q(2)-GRS) CoMFA (see ref 1) was employed, as were two novel variable selection QSAR methods recently developed in one of our laboratories. These latter methods included genetic algorithm-partial least squares (GA-PLS) and K nearest neighbor (KNN) procedures (see refs 2-4), which utilize 2D topological descriptors of chemical structures. Each QSAR approach resulted in a highly predictive model, with cross-validated R(2) (q(2)) values of 0.57 for CoMFA, 0.54 for q(2)-GRS, 0.73 for GA-PLS, and 0.79 for KNN. The success of all of the QSAR methods indicates the presence of an intrinsic structure-activity relationship in this group of compounds and affords more robust design and prediction of biological activities of novel D(1) ligands. PMID- 10464010 TI - Synthesis, in vitro pharmacology, and molecular modeling of very potent tacrine huperzine A hybrids as acetylcholinesterase inhibitors of potential interest for the treatment of Alzheimer's disease. AB - Eleven new 12-amino-6,7,10,11-tetrahydro-7, 11-methanocycloocta[b]quinoline derivatives [tacrine (THA)-huperzine A hybrids, rac-21-31] have been synthesized as racemic mixtures and tested as acetylcholinesterase (AChE) inhibitors. For derivatives unsubstituted at the benzene ring, the highest activity was obtained for the 9-ethyl derivative rac-20, previously prepared by our group. More bulky substituents at position 9 led to less active compounds, although some of them [9 isopropyl (rac-22), 9-allyl (rac-23), and 9-phenyl (rac-26)] show activities similar to that of THA. Substitution at position 1 or 3 with methyl or fluorine atoms always led to more active compounds. Among them, the highest activity was observed for the 3-fluoro-9-methyl derivative rac-28 [about 15-fold more active than THA and about 9-fold more active than (-)-huperzine A]. The activity of some THA-huperzine A hybrids (rac-19, rac-20, rac-28, and rac-30), which were separated into their enantiomers by chiral medium-pressure liquid chromatography (chiral MPLC), using microcrystalline cellulose triacetate as the chiral stationary phase, showed the eutomer to be always the levorotatory enantiomer, their activity being roughly double that of the corresponding racemic mixture, the distomer being much less active. Also, the activity of some of these compounds inhibiting butyrylcholinesterase (BChE) was tested. Most of them [rac 27-31, (-)-28, and (-)-30], which are more active than (-)-huperzine A as AChE inhibitors, turned out to be quite selective for AChE, although not so selective as (-)-huperzine A. Most of the tested compounds 19-31 proved to be much more active than THA in reversing the neuromuscular blockade induced by d tubocurarine. Molecular modeling of the interaction of these compounds with AChE from Torpedo californica showed them to interact as truly THA-huperzine A hybrids: the 4-aminoquinoline subunit of (-)-19 occupies the same position of the corresponding subunit in THA, while its bicyclo[3.3.1]nonadiene substructure roughly occupies the same position of the corresponding substructure in (-) huperzine A, in agreement with the absolute configurations of (-)-19 and (-) huperzine A. PMID- 10464012 TI - New 4-point pharmacophore method for molecular similarity and diversity applications: overview of the method and applications, including a novel approach to the design of combinatorial libraries containing privileged substructures. AB - A new 4-point pharmacophore method for molecular similarity and diversity that rapidly calculates all potential pharmacophores/pharmacophoric shapes for a molecule or a protein site is described. The method, an extension to the ChemDiverse/Chem-X software (Oxford Molecular, Oxford, England), has also been customized to enable a new internally referenced measure of pharmacophore diversity. The "privileged" substructure concept for the design of high-affinity ligands is presented, and an example of this new method is described for the design of combinatorial libraries for 7-transmembrane G-protein-coupled receptor targets, where "privileged" substructures are used as special features to internally reference the pharmacophoric shapes. Up to 7 features and 15 distance ranges are considered, giving up to 350 million potential 4-point 3D pharmacophores/molecule. The resultant pharmacophore "key" ("fingerprint") serves as a powerful measure for diversity or similarity, calculable for both a ligand and a protein site, and provides a consistent frame of reference for comparing molecules, sets of molecules, and protein sites. Explicit "on-the-fly" conformational sampling is performed for a molecule to enable the calculation of all geometries accessible for all combinations of four features (i.e., 4-point pharmacophores) at any desired sampling resolution. For a protein site, complementary site points to groups displayed in the site are generated and all combinations of four site points are considered. In this paper we report (i) the details of our customized implementation of the method and its modification to systematically measure 4-point pharmacophores relative to a "special" substructure of interest present in the molecules under study; (ii) comparisons of 3- and 4-point pharmacophore methods, highlighting the much increased resolution of the 4-point method; (iii) applications of the 4-point potential pharmacophore descriptors as a new measure of molecular similarity and diversity and for the design of focused/biased combinatorial libraries. PMID- 10464011 TI - 5'-Phosphoramidates and 5'-diphosphates of 2'-O-allyl-beta-D arabinofuranosyluracil, -cytosine, and -adenine: inhibition of ribonucleotide reductase. AB - Continuing our studies on ribonucleotide reductase (RNR) mechanism-based inhibitors, we have now prepared the diphosphates (DP) of 2'-O-allyl-1-beta-D arabinofuranosyl-uracil and -cytosine and 2'-O-allyl-9-beta-D-arabinofuranosyl adenine and evaluated their inhibitory activity against recombinant murine RNR. 2'-O-Allyl-araUDP proved to be inhibitory to RNR at an IC(50) of 100 microM, whereas 2'-O-allyl-araCDP was only marginally active (IC(50) 1 mM) and 2'-O-allyl araADP was completely inactive. The susceptibility of the parent nucleosides to phosphorylation by thymidine kinase and 2'-deoxycytidine kinase was also investigated, and all nucleosides proved to be poor substrates for the above cited kinases. Moreover, prodrugs of 2'-O-allyl-araU and -araC monophosphates, namely 2'-O-allyl-5'-(phenylethoxy-L-alanyl phosphate)-araU and -araC, were prepared and tested against tumor cell proliferation but proved to be inactive. A molecular modeling study has been conducted in order to explain our results. The data confirm that for both the natural and analogue nucleoside diphosphates, the principal determinant interaction with the active site of RNR is with the diphosphate group, which forms strong hydrogen bonds with Glu623, Thr624, Ser625, and Thr209. Our findings indicate that the poor phosphorylation may represent an explanation for the lack of marked in vitro cytostatic activity of the test compounds. PMID- 10464013 TI - Novel antidiabetic and hypolipidemic agents. 5. Hydroxyl versus benzyloxy containing chroman derivatives. AB - Several thiazolidinediones having chroman moieties were synthesized and evaluated for their euglycemic and hypolipidemic activities. Some of the analogues having an aminoalkyl group as a linker between the chroman ring and 4-[5-(2,4-dioxo-1, 3 thiazolidinyl)methyl]phenoxy moiety seem to be better than troglitazone. In vitro transactivation assays of PPARgamma have been carried out with these glitazones to understand their molecular mechanism. For the first time we have found that some of the unsaturated thiazolidinediones are superior to their saturated counterpart in the in vivo assay. A more potent thiazolidinedione analogue than troglitazone is reported. Pharmacokinetic studies have shown that protection of the OH group in the chroman moiety leads to a decrease in metabolism, thereby resulting in a superior pharmacological profile. PMID- 10464014 TI - Novel arylpyrazino[2,3-c][1,2,6]thiadiazine 2,2-dioxides as platelet aggregation inhibitors. 2. Optimization by quantitative structure-activity relationships. AB - In the previous paper (Part 1), we described the synthesis and antiplatelet activity of a series of phenyl- and heteroarylpyrazino[2,3-c][1,2,6]thiadiazine 2,2-dioxides. In this paper, we report the optimization of the platelet aggregation inhibitory activity by an iterative sequence of quantitative structure-activity relationship studies which encompassed synthesis and evaluation of the effects of structure variations at the 1-, 6-, and 7-positions of the heterocyclic system. A model has been established that correctly correlates antiplatelet activity in this series with the partial atomic charges calculated by a local density functional ab initio method. As a result of this study, the experimental platelet aggregation inhibitory activity of the lead compound was improved 300-fold. PMID- 10464015 TI - Structural modification of Fas C-terminal tripeptide and its effects on the inhibitory activity of Fas/FAP-1 binding. AB - We report the structural requirements of the C-terminal tripeptide derivative of Fas (Ac-Ser-Leu-Val-OH, 1) for the inhibitory activity of Fas/FAP-1 binding. The presence of a carboxyl group and a L-Val residue at the C-terminus is essential for the inhibitory activity, and the hydroxyl group of Ser plays an important role as the donor of a hydrogen bond. The introduction of hydrophobic groups to the N-terminal region of 1, especially the phenylaminocarbonyl group (41), showed a remarkable increase in potency. Further improvement was observed by the attachment of the Glu residue to the meta-position of the phenyl ring of 41 (51). The ester derivative of 41 (56) had the ability to induce apoptosis which was dependent on the concentration of anti-Fas antibody in the colon cancer cell line, DLD-1, which expresses both Fas and FAP-1 and is resistant to Fas-induced apoptosis. We are now investigating whether FAP-1 is a main target of 56 and whether the inhibition of Fas/FAP-1 binding by 56 retrieves the apoptotic signal. PMID- 10464016 TI - Synthesis and properties of 3-(2-hydroxyethyl)-3-n-pentyldiazirine, a photoactivable general anesthetic. AB - To overcome the difficulties of locating the molecular sites of general anesthetic action, we synthesized a novel photoactivable general anesthetic, 3-(2 hydroxyethyl)-3-n-pentyldiazirine (3-diazirinyloctanol), which anesthetized tadpoles with an ED(50) of 160 microM. Subanesthetic concentrations of 3 diazirinyloctanol enhanced GABA-induced currents in GABA(A) receptors, an effect that has been implicated in general anesthetic action. It also enhanced [(3)H]muscimol binding to this receptor. In muscle nicotinic acetylcholine receptors (nAcChoR), it inhibited the response to acetylcholine with an IC(50) of 33 microM. 3-Diazirinyloctanol's pharmacological actions were comparable to those of octanol. 3-(2-Hydroxyethyl)-3-[4,5-(3)H(2)]-n-pentyldiazirine photoincorporated into Torpedo nAcChoR-rich membranes mainly in the alpha subunit with 70% being in a proteolytic fragment containing the M4 transmembrane segment. Agonist enhanced the photolabeling 10-fold in a fragment containing the M1, M2, and M3 transmembrane segments. Thus, 3-diazirinyloctanol is a novel general anesthetic that acts on, and can be photoincorporated into, postsynaptic receptors. PMID- 10464017 TI - Virtual combinatorial syntheses and computational screening of new potential anti herpes compounds. AB - The activity of new anti-HSV-1 chemical structures, designed by virtual combinatorial chemical synthesis and selected by a computational screening, is determined by an in vitro assay. A virtual library of phenol esters and anilides was formed from two databases of building blocks: one with carbonyl fragments and the other containing both substituted phenoxy and phenylamino fragments. The library of virtually assembled compounds was computationally screened, and those compounds which were selected by our mathematical model as active ones were finally synthesized and tested. Our antiviral activity model is a "tandem" of four linear functions of topological graph-theoretical descriptors. A given chemical structure was selected as active if it satisfies every discriminant equation in that model. The final result was that five new structures were selected, synthesized, and tested: all of them demonstrated activity, and three showed appreciable anti-HSV-1 activity, with IC(50) values of 0.9 microM. The same model, applied to a database of known compounds, has identified the anti herpes activity of the following compounds: 3,5-dimethyl-4-nitroisoxazole, nitrofurantoin, 1-(pyrrolidinocarbonylmethyl)piperazine, nebularine, cordycepin, adipic acid, thymidine, alpha-thymidine, inosine, 2, 4-diamino-6 (hydroxymethyl)pteridine, 7-(carboxymethoxy)-4-methylcoumarin, 5-methylcytidine, and others that showed less activity. PMID- 10464018 TI - Synthesis and evaluation of 3-trifluoromethyl-7-substituted-1,2,3, 4 tetrahydroisoquinolines as selective inhibitors of phenylethanolamine N methyltransferase versus the alpha(2)-adrenoceptor. AB - A series of 3-trifluoromethyl-1,2,3,4-tetrahydroisoquinolines was synthesized and evaluated as inhibitors of phenylethanolamine N-methyltransferase (PNMT) and as inhibitors of the binding of clonidine at the alpha(2)-adrenoceptor. These compounds were found to be selective inhibitors of PNMT due to their decreased affinity for the alpha(2)-adrenoceptor, which was attributed to steric bulk intolerance around the 3-position of 1,2,3,4-tetrahydroisoquinoline (THIQ) at the alpha(2)-adrenoceptor and to the decreased pK(a) of the THIQ amine due to the 3 trifluoromethyl moiety. Overall, these compounds displayed less affinity for PNMT compared to previously studied THIQ-type inhibitors, except for 16 which was found to have good affinity for PNMT (PNMT K(i) = 0.52 microM). Compounds 14 and 16 proved to be the most selective inhibitors in this small series of compounds and are some of the most selective inhibitors of PNMT known (14, selectivity alpha(2) K(i)/PNMT K(i) = 700; 16, selectivity alpha(2) K(i)/PNMT K(i) > 1900). Compounds 14 and 16 are also quite lipophilic due to the 3-trifluoromethyl moiety and represent promising new leads for the development of new highly selective inhibitors of PNMT, which should be sufficiently lipophilic to penetrate the blood-brain barrier. PMID- 10464019 TI - Design of highly active analogues of the pyrrolo[1,2-a]benzimidazole antitumor agents. AB - The pyrrolo[1,2-a]benzimidazole (PBI) reductive alkylating agents have been investigated in this laboratory since their discovery in the late 1980s. Of all the structural modifications of the PBIs investigated so far, the variation of the 3-substituent has the greatest influence on cytotoxicity, toxicity, and in vivo antitumor activity. In the present study, we prepared both the R and S enantiomers of nitrogen-containing 3-substituents possessing hydrogen-bonding capability as well as varying basicity. The rationale was to take advantage of stereoselective DT-diaphorase reductive activation as well as hydrogen bonding in the DNA major groove. As a result of these studies, analogues were discovered possessing among the highest hollow fiber tumor assay scores observed in hundreds of natural and synthetic antitumor agents. Our findings indicate that a relatively basic 3-substituent is required for outstanding PBI cytotoxicity but that the importance of using pure enantiomers is still open to study. PMID- 10464020 TI - Thiazolothiazepine inhibitors of HIV-1 integrase. AB - A series of thiazolothiazepines were prepared and tested against purified human immunodeficiency virus type-1 integrase (HIV-1 IN) and viral replication. Structure-activity studies reveal that the compounds possessing the pentatomic moiety SC(O)CNC(O) with two carbonyl groups are in general more potent against purified IN than those containing only one carbonyl group. Substitution with electron-donating or -withdrawing groups did not enhance nor abolish potency against purified IN. By contrast, compounds with a naphthalene ring system showed enhanced potency, suggesting that a hydrophobic pocket in the IN active site might accommodate an aromatic system rather than a halogen. The position of sulfur in the thiazole ring appears important for potency against IN, as its replacement with an oxygen or carbon abolished activity. Further extension of the thiazole ring diminished potency. Compounds 1, 19, and 20 showed antiviral activity and inhibited IN within similar concentrations. These compounds inhibited IN when Mn(2+) or Mg(2+) was used as cofactor. None of these compounds showed detectable activities against HIV-1 reverse transcriptase, protease, virus attachment, or nucleocapsid protein zinc fingers. Therefore, thiazolothiazepines are potentially important lead compounds for development as inhibitors of IN and HIV replication. PMID- 10464021 TI - N-Substituted (2,3-dihydro-1,4-benzodioxin-2-yl)methylamine derivatives as D(2) antagonists/5-HT(1A) partial agonists with potential as atypical antipsychotic agents. AB - A series of N-substituted 1-(2,3-dihydro-1, 4-benzodioxin-2-yl)methylamine derivatives with D(2) antagonist/5-HT(1A) partial agonist activity has been prepared as potential atypical antipsychotic agents. Optimization of in vitro receptor binding activity and in vivo activity in rodent models of psychosis has led to compound 24, which showed good affinities for human D(2), D(3), and 5 HT(1A) receptors but significantly less affinity for human alpha(1) adrenoceptors and rat H(1) and muscarinic receptors. In rodents, 24 showed functional D(2)-like antagonism and 5-HT(1A) partial agonism. After oral dosing, 24 showed good activity in rodent antipsychotic tests and very little potential to cause extrapyramidal side effects (EPS), as measured by its ability to induce catalepsy in rats only at very high doses. In the light of this promising profile of activity, 24 has been selected for clinical investigation as a novel antipsychotic agent with a predicted low propensity to cause EPS. PMID- 10464022 TI - Design and in vivo analysis of potent non-thiol inhibitors of farnesyl protein transferase. AB - Inhibitors of farnesyl protein transferase (FPTase) based upon a pseudotripeptide template are described that comprise an imidazole group substituted with a hydrophobic substituent. (1, 5)-Disubstitution of the imidazole group is shown to be the optimal array that leads to potent and selective inhibitors of FPTase. A variety of aryl and isoprenyl substituents are shown to afford effective inhibitors, and the mechanism by which these compounds inhibit FPTase has been investigated. The biochemical behavior of these compounds suggests that they bind to FPTase at the site usually occupied by the protein substrate. In experiments in cell culture, the methyl ester prodrugs of these inhibitors are cell permeant and potently inhibit the posttranslational modification of H-Ras protein. Additionally, these molecules revert the phenotype of ras transformed cells as evidenced by their ability to slow the growth of ras transformed cell lines in soft agar. One of the inhibitors, as its methyl prodrug, was evaluated in two in vivo models of tumor growth. The compound selectively inhibited the growth of tumors derived from H-ras transformed cells, in nude mice, and caused the regression of preexisting tumors in an H-ras transgenic animal model. PMID- 10464023 TI - Probing the topological arrangement of the N- and C-terminal residues of bradykinin for agonist activity at the B1 receptor. AB - The conformational features of H-Lys-Arg-Ado-Ser-Pro-Phe-OH (Ado = 12 aminododecanoic acid), a des-Arg(9) analogue of Lys-bradykinin, have been determined by high-resolution NMR in the presence of a zwitterionic lipid environment. The analogue is the most active member of a series of analogues designed to probe the topological arrangement of the N- and C-termini required for agonistic activity at the B1 kinin receptor. A novel computational procedure for the utilization of NOE constraints from cis and trans configurational isomers is illustrated. Only with this computational methodology could the structural features of the N-terminus of the peptide be determined. Using radical-induced relaxation of the (1)H NMR signals, we measured the topological orientation of the peptide with respect to the zwitterionic lipid interface. The results indicate that the long, alkyl chain of the Ado amino acid imbeds into the lipid surface. The structural features of the C-terminus of the B1-selective analogue consist of a well-defined turn. Although removed from a standard beta-turn, required for activity at the B2 kinin receptor, the topological orientation of the side chains of the des-Arg(9) compound are surprisingly similar to those previously observed for beta-turn-containing bradykinin analogues. Therefore, we attribute the high B1 receptor selectivity, observed upon removal of Arg(9) from bradykinin, solely to the loss of a charged amino acid and not to altered structural features. PMID- 10464024 TI - Synthesis of 5-(carboranylalkylmercapto)-2'-deoxyuridines and 3 (carboranylalkyl)thymidines and their evaluation as substrates for human thymidine kinases 1 and 2. AB - Derivatives of thymidine containing o-carboranylalkyl groups at the N-3 position and derivatives of 2'-deoxyuridine containing o-carboranylalkylmercapto groups at the C-5 position were synthesized. The alkyl spacers consist of 4-8 methylene units. The synthesis of the former compounds required 3-4 reaction steps in up to 75% overall yield and that of the latter 9-10 reaction steps with significantly lower overall yield. Derivatives of thymidine substituted with carboranylalkyl substituents at the N-3 position and short spacers were phosphorylated by both recombinant and purified cytosolic thymidine kinase (TK1) to a relatively high degree. None of the tested 2'-deoxyuridine derivatives possessing carboranyl substituents at the C-5 position were phosphorylated by either recombinant or purified TK1. The amounts of phosphorylation product detected for some of the C-5 substituted nucleosides with recombinant mitochondrial thymidine kinase (TK2) were low but significant and decreased with increasing lengths of the alkyl spacer. The data obtained in this study do not seem to support the tether concept applied in the synthesis of the new C-5- and N-3-substituted carboranyl nucleosides intended to reduce possible steric interference in the binding of carboranyl nucleosides with deoxynucleoside kinases. Instead, it appeared that a closer proximity of the bulky carborane moiety to the nucleoside scaffold resulted in better substrate characteristics. PMID- 10464025 TI - Asymmetric synthesis and antiviral activities of L-carbocyclic 2', 3'-didehydro 2',3'-dideoxy and 2',3'-dideoxy nucleosides. AB - Asymmetric syntheses of L-carbocyclic 2',3'-didehydro-2',3'-dideoxy- and 2',3' dideoxypyrimidine and purine nucleoside analogues were accomplished, and their anti-HIV and anti-HBV activities were evaluated. The key intermediate, (1S, 4R)-1 benzoyloxy-4-(tert-butoxymethyl)cyclopent-2-ene (7), was prepared by benzoylation of the alcohol 2, selective deprotection of the isopropylidene group of 3, followed by thermal elimination via cyclic ortho ester or deoxygenation via cyclic thionocarbonate. The target compounds were also synthesized by thermal elimination via cyclic ortho esters from protected nucleosides. It was found that L-carbocyclic 2',3'-didehydro-2',3'-dideoxyadenosine (34) exhibited potent anti HBV activity (EC(50) = 0.9 microM) and moderate anti-HIV activity (EC(50) = 2.4 microM) in vitro without cytotoxicity up to 100 microM. PMID- 10464026 TI - 5-Amino-1-(chloromethyl)-1,2-dihydro-3H-benz[e]indoles: relationships between structure and cytotoxicity for analogues bearing different DNA minor groove binding subunits. AB - A series of 5-amino-seco-CBI compounds, designed for use as effectors for prodrugs, were prepared to study structure-activity relationships for the cytotoxicity of side chain analogues. Compounds were prepared by coupling 1 (chloromethyl)-5-nitro-1, 2-dihydro-3H-benz[e]indole to appropriate carboxylic acids, followed by nitro group reduction, or by coupling suitable 5-amino protected indolines to alpha,beta-unsaturated acids, followed by deblocking. These AT-specific DNA alkylating agents were evaluated for cytotoxicity in a series of tumor cell lines (AA8, UV4, EMT6, SKOV3). For those analogues bearing an indolecarbonyl side chain, the 5'-methoxy derivative was the most cytotoxic (IC(50) 1.3 nM in AA8 cells, 4 h exposure), comparable to that of the parent CBI TMI (5', 6',7'-trimethoxyindole) derivative (IC(50) 0.46 nM in the above assay). A subset of solubilized derivatives bearing O(CH(2))(2)NMe(2) substituents were about 10-fold less potent. For compounds containing an acryloyl linker in the side chain, the 4'-methoxycinnamoyl derivative proved the most cytotoxic (IC(50) 0. 09 nM in the above assay). A number of these 5-amino-seco-CBI-TMI analogues (including the solubilized compounds) are of interest both as cytotoxins and as components of amine-based prodrugs designed for tumor-specific activation. PMID- 10464028 TI - Dipole Information Complementarity in Discrete 2D Patterns. AB - An information theoretic approach is used to derive equations describing the dipole information in discrete two-dimensional (2D) patterns. Dipoles are pair wise, or second-order, interactions of pattern states. It is shown that the basic dipole information equation can be decomposed into two complementary representations of the total information. One of these representations expresses the total information in terms of the sum of two components: the dipole spatial information and the permutation set information. The spatial information component describes the information in the distribution of spatial transitions for each type of color transition. The permutation set information component is a second-order function of the single color probabilities. The second representation is also the sum of two components: the dipole color information and the array information. The dipole color information describes the information in the distribution of color transitions for each possible spatial transition. The array information is a function of the size and geometry of the pattern array. These two representations characterize the total information from complementary perspectives, each emphasizing different aspects of the pattern's structure. Implications of this approach for the understanding of patterns and pattern perception are examined. Copyright 1999 Academic Press. PMID- 10464027 TI - Structure-activity relationship of N-(phenylalkyl)cinnamides as novel NR2B subtype-selective NMDA receptor antagonists. AB - A novel series of N-(phenylalkyl)cinnamides related to N-(4-phenylbutyl)-3,4 dihydroxy-beta-cyanocinnamide (6, an EGFR-K inhibitor with high antiproliferative activity) was synthesized and tested for antagonism at N-methyl-D-aspartate (NMDA) receptor subtypes. Potency and subunit selectivity were assayed by electrical recordings in Xenopus oocytes expressing three binary combinations of cloned rat NMDA receptor subunits: NR1A expressed in combination with either NR2A, NR2B, or NR2C. The N-(phenylalkyl)cinnamides are selective antagonists of NR1A/2B receptors. Assayed under steady-state conditions, N-(4-phenylbutyl)-4 hydroxycinnamide (16) has an IC(50) value of 77 nM and >1000-fold selectivity with respect to NR1A/2A and NR1A/2C receptors. Potency at alpha(1) adrenergic receptors is low for the four cinnamides tested. Inhibition of NR1A/2B receptors does not correlate with EGFR and ErbB2/neu tyrosine kinase inhibitor activity. The N-(phenylalkyl)cinnamide series we describe provides a novel and structurally diverse framework for designing new NR2B-selective NMDA antagonists as potential CNS therapeutics. PMID- 10464029 TI - A Dimension-Related Metric on the Lattice of Knowledge Spaces. AB - The set of all knowledge spaces on a given set of items or questions is investigated with respect to order theoretic and metrical properties. It is argued that a generalization of properties of the subspaces of a finite dimensional Euclidean vector space yields the material for defining a metric which satisfies certain requirements. The fact that the lattice of knowledge spaces is not modular is the reason for most of the difficulties. It turns out that a Hausdorff metric based on the Hamming distance satisfies the postulated conditions. Copyright 1999 Academic Press. PMID- 10464030 TI - A Mental Space Similarity Group Model of Shape Constancy. AB - A group model of mental transformations based on the geometric model of P. B. Yale (1968, Geometry and symmetry, Holden-Day, San Francisco) was constructed for form recognition. The model consisted of nine characteristic subgroups of the similarity group in Euclidean space. With these subgroups, six series were formed, representing six visual paths for form recognition. Each series involved five characteristic subgroups. Six subframes were associated with nine characteristic subgroups in the model. These subframes were shape (angle measure), the sense, size (volume), verticality, uprightness, and position. The model was validated by an experiment, using reaction time as the behavior index. Since shape is the common invariant property of all subgroups of the similarity group, angle measure was not included in ordering of subframes. The findings show that the preservation of uprightness of a form provides the best condition for form recognition, followed by the preservation of sense and verticality of a form. While the effect of position is not strong, size has the weakest influence on space form recognition. Copyright 1999 Academic Press. PMID- 10464031 TI - Set-Valued Utilities for Strict Partial Orders. AB - So far, only weak utility functions have been used to represent strict partial orders, but such a representation is not completely satisfactory, since we can not reconstruct the preference relation by simply knowing the weak utility function. The purpose of this paper is to present a type of numerical representation for strict partial orders, in such a way that recovering the preference relation is possible. Taking into account that (2(ℝ), superset) is a strict partial order, we consider set-valued utilities, namely, set-valued mappings, such that they admit a selection which is a weak utility function. Copyright 1999 Academic Press. PMID- 10464032 TI - TELEGRAPHIC REVIEWS. PMID- 10464033 TI - Plasmodium falciparum: detection of a novel asparagine-rich protein on the surface of sporozoite. AB - We had previously cloned and characterized a gene for a novel asparagine-rich protein from P. falciparum (PfARP), a target of natural human immune response. The antibodies to PfARP were localized to the surface of parasitized red blood cells and reacted with intracellular components in all erythrocytic asexual and sexual stages of the parasite. We here describe reactivity of antibodies against this novel PfARP on the surface of mosquito stage sporozoite of P. falciparum by indirect immunofluorescence assay and immunoelectron microscopy, the latter revealing a highly periodic punctate pattern of distribution of PfARP on the surface of sporozoite. These results suggest a possibility that PfARP might represent yet another sporozoite surface protein. PMID- 10464034 TI - Leishmania: fine mapping of the Leishmanolysin molecule's conserved core domains involved in binding and internalization. AB - The Leishmanolysin molecule's role in the uptake of Leishmania parasites by the human U937 pro-myelocytic cell line was studied, using synthetic peptides representing the complete Leishmania (Viannia) guyanensis Leishmanolysin protein amino acid sequence. The particular peptides present in two protein's core domains efficiently impaired the internalization of promastigotes from four different Leishmania species and modified the kinetics of the binding of heterologous recombinant Leishmanolysin protein. The functional domains which exhibited this property represent a highly conserved portion of the sequence among different Leishmania species. The peptides' inhibitory activity correlated with their ability to bind molecules present on the surface of the human cell line. One of the two functional core domains identified involves the previously described adhesive sequence (SRYD) and the putative zinc-binding motif (HExxH). The second functional core domain includes a third histidine residue coordinated with zinc which determines the molecule's structural features. These findings indicate that the molecular interactions between Leishmanolysin's conserved domains and the macrophage surface molecules efficiently contribute to the parasite's internalization. Induction of neutralizing immune responses, which impair the early parasite-host interaction described here, may be an important alternative in designing synthetic subunit human leishmaniasis vaccines. PMID- 10464035 TI - Toxoplasma gondii in primary rat CNS cells: differential contribution of neurons, astrocytes, and microglial cells for the intracerebral development and stage differentiation. AB - The central nervous system (CNS) of the intermediate host plays a central role in the lifelong persistence of Toxoplasma gondii as well as in the pathogenesis of congenital toxoplasmosis and reactivated infection in immunocompromised patients. In order to analyze the parasite-host interaction within the CNS, the host cell invasion, the intracellular replication, and the stage conversion from tachyzoites to bradyzoites was investigated in mixed cultures of dissociated CNS cells from cortices of Wistar rat embryos. Two days post infection (p.i.) with T. gondii tachyzoites, intracellular parasites were detected within neurons, astrocytes, and microglial cells as assessed by double immunofluorescence and confocal microscopy. Quantitative analyses revealed that approximately 10% of neurons and astrocytes were infected with T. gondii, while 30% of the microglial cells harbored intracellular parasites. However, the replication of T. gondii within microglial cells was considerably diminished, since 93% of the parasitophorous vacuoles (PV) contained only one to two parasites which often appeared degenerated. This toxoplasmacidal activity was not abrogated after treatment with NO synthase inhibitors or neutralization of IFN-gamma production. In contrast, 30% of the PV in neurons and astrocytes harbored clearly proliferating parasites with at least four to eight parasites per vacuole. Four days p.i. with tachyzoites of T. gondii, bradyzoites were detected within neurons, astrocytes, and microglial cells of untreated cell cultures. However, the majority of bradyzoite-containing vacuoles were located in neurons. Spontaneous differentiation to the bradyzoite stage was not inhibited after addition of NO synthase inhibitors or neutralization of IFN-gamma. In conclusion, our results indicate that intracerebral replication of T. gondii as well as spontaneous conversion from the tachyzoite to the bradyzoite stage is sustained predominantly by neurons and astrocytes, whereas microglial cells may effectively inhibit parasitic growth within the CNS. PMID- 10464036 TI - Plasmodium falciparum: drug-resistant malaria complicating leukemias and lymphomas in children. AB - The present communication deals with drug-resistant Plasmodium falciparum malaria complicating hematologic malignancies (leukemias, n = 24, and lymphomas, n = 7) in children. Of 50 cases of hematologic malignancies, 31 patients were microscopically diagnosed as having P. falciparum infection (MP +). Initially, all the patients were treated with chloroquine. The results of primary treatment showed chloroquine resistance in 16 (51. 62%) cases. Of these 16 chloroquine resistant cases, 13 were secondarily treated with a combination of pyrimethamine plus sulfamethopyrazine. The results of secondary treatment also revealed resistance to pyrimethamine plus sulfamethopyrazine in 6 of 13 (46. 10%) cases. The 6 pyrimethamine plus sulfamethopyrazine-resistant P. falciparum cases were finally cured by quinine therapy, against which no resistance was encountered. Conversely, in the control group comprising 38 cases of P. falciparum without malignancy, the incidence of chloroquine resistance was found in only 9 cases, which is rather low (23.70%). Of these 7 chloroquine-resistant cases, 5 were found to be sensitive to pyrimethamine plus sulfamethopyrazine treatment, while the 2 nonresponders were finally cured with quinine. The overall results of this study show a high prevalence of chloroquine resistance among clinical cases of falciparum malaria (25/69; 30.6%). Among the nonresponders (n = 20) 40% of cases were also resistant to the pyrimethamine plus sulfamethopyrazine combination. There was no resistance to quinine. PMID- 10464037 TI - Trypanosoma cruzi: conformational preferences of antigenic peptides bearing the immunodominant epitope of the B13 antigen. AB - The Trypanosoma cruzi recombinant protein B13 contains tandemly repeated domains and shows high sensitivity in the serological diagnosis of Chagas' disease. It has been shown that the immunodominant epitope of B13 is contained in the GDKPSLFGQAAAGDKPSLF-NH(2) sequence and that the hexapeptide AAAGDK seems to be the "core" of that epitope. Three peptides containing that "core" sequence, one corresponding to the entire repeat motif GDKPSLFGQAAAGDKPSLF-NH(2), pB13, and two smaller fragments, FGQAAAGDK-NH(2), S4, and QAAAGDKPS-NH(2), S5, have been tested in competitive ELISA with recombinant protein B13 in the solid phase against 40 chagasic sera from Brazilian patients. The median percentage inhibition for pB13, S4, and S5 were, respectively, 91, 86, and 68%. The possibility that the distinct antigenic activity of those peptides correlates with the existence of preferential conformational properties has been investigated by CD and NMR spectroscopy. Results indicate their propensity to adopt a helical configuration, centered in the AAAGDK sequence, and whose extent and stability directly correlates with the peptides' antigenicity. The data are discussed in the light of the existence of conformational preferences involving immunodominant epitopes in tandemly repeated antigens. PMID- 10464038 TI - Plasmodium chabaudi: effect of antimalarial drugs on gametocytogenesis. AB - The proportion of asexual blood-stage malaria parasites that develop into transmission stages (gametocytes) can increase in response to stress. We investigated whether stress imposed by a variety of antimalarial drugs administered before or during infection increased gametocyte production (gametocytogenesis) in vivo in the rodent malaria parasite, Plasmodium chabaudi. All methods of drug treatment greatly reduced the numbers of asexual parasites produced during an infection but resulted in either no reduction in numbers of gametocytes or a smaller reduction than that experienced by asexuals. We used a simple model to estimate temporal variation in gametocyte production. Temporal patterns of gametocytogenesis did not greatly differ between untreated and prophylaxis infections, with rates of gametocytogenesis always increasing as the infection progressed. In contrast, administration of drugs 5 days after infection stimulated increased rates of gametocytogenesis early in the infection, resulting in earlier peak gametocyte densities relative to untreated infections. Given the correlation between gametocyte densities and infectivity to mosquito vectors, and the high frequency of subcurative drug therapy and prophylaxis in human populations, these data suggest that antimalarial drugs may frequently have only a small effect on reducing malaria transmission and may help to explain the rapid spread of drug-resistant geno-types. PMID- 10464039 TI - T cells facilitate Brugia malayi development in TCRalpha(null) mice. AB - The host-parasite interactions of Brugia malayi in mice are complex and multifactorial. In order to study the role of T cells in early B. malayi development, we infected TCRalpha(null) mice, which retain a population of CD4+ TCRbeta+ cells and TCRbeta(null) mice, which lack all TCRalphabeta(+) T cells. TCRalpha(null) mice were permissive to L4 larval and adult worm development but TCRbeta(null) mice were not. Depletion of the CD4(+) T cells in the former abrogated the permissive phenotype. It appears that the CD4(+) TCRbeta(+) T cells that have been described in TCRalpha(null) mice may facilitate early B. malayi development. These data are similar to our earlier demonstration of the role of NK cells in facilitating worm growth in SCID mice. PMID- 10464040 TI - Plasmodium cynomolgi: transfection of blood-stage parasites using heterologous DNA constructs. PMID- 10464041 TI - Lifestyle determinants of cancer among Danish mastic asphalt workers. AB - Three reports suggest that asphalt workers, especially young mastic asphalt workers, in Denmark experience an increase in incidence and mortality from cancer and in mortality from other conditions. The methodology described in these reports raises questions about their validity and the data presented are limited and difficult to interpret. The cancers and the causes of death that are increased are highly correlated with those seen among men in the general population who use alcohol in excess, who smoke, and who engage in other risk taking behaviors. The effects of these lifestyle causes of disease were largely not controlled in the reported studies. These behaviors, which cluster in young men, rather than exposure to asphalt fumes, probably caused the disease patterns that were reported. Policy makers who use epidemiologic results for risk assessment and regulation should do so with care. Working men and women sometimes die at high rates and their occupational exposures may or may not be responsible. The distinction is crucial if occupational health is to be improved. If asphalt workers die young from excessive drinking and smoking, we are not protecting their health by controlling asphalt fume exposures. PMID- 10464042 TI - Review of subchronic/chronic toxicity of antimony potassium tartrate. AB - Subchronic/chronic toxicity studies on antimony potassium tartrate (APT) have been reviewed. One of the older studies (H. A. Schroeder et al., 1970, J. Nutr. 100 (1), 59-68), on which are based the EPA reference dose value and a number of state, national, and international drinking water criteria for antimony, has severe inadequacies in study conduct making it uninterpretable and inappropriate for characterization of APT toxicity. In particular, the manner in which control data were generated and utilized in this study is considered invalid. More recent drinking water studies conducted by the NTP (1992, "NTP Technical Report on Toxicity Studies of Antimony Potassium Tartrate in F344/N Rats and B6C3F(1) Mice (Drinking Water and Intraperitoneal Injection Studies)," NTP Toxicity Report Series, No. 11) and Poon et al. (1998, Food Chem. Toxicol. 36, 20-35) showed antimony to be of low toxicity. The NOAEL in the 14-day NTP study was 2500 ppm by the oral route in both rats and mice, while Poon et al. (1998) suggested a NOAEL of 0.5 ppm in their 90-day study. However, upon close examination, it was determined that this value was based on subtle histological changes in the thyroid gland that were physiological, not toxicological, in nature. This conclusion is supported further by an absence of these changes in a well conducted 13-week intraperitoneal exposure study in rats that utilized APT at much higher doses (NTP, 1992). Thus, the NOAEL by Poon et al. (1998) should more appropriately be 50 ppm. When regulatory criteria for antimony are established and/or reviewed, the findings in the NTP study and this critical reevaluation of the Poon et al. (1998) study should be considered when establishing a NOAEL for subchronic exposure to antimony in the future. PMID- 10464043 TI - Modeling of deposition and clearance of inhaled Ni compounds in the human lung. AB - By extrapolation from the rat study, a mathematical model of deposition, clearance, and retention kinetics for inhaled Ni compounds (high-temperature (green) NiO, Ni(3)S(2), and NiSO(4). 6H(2)O) in the alveolar region of the human lung has been developed. For human deposition, an updated version of an earlier model (C. P. Yu and C. K. Diu, 1982, Am. Ind. Hyg. Assoc. J.) was used in this study. Because of the profound differences in physiological and ventilation conditions between humans and rats, humans were found to have a higher alveolar deposition fraction than rats when exposed to the same Ni compounds. However, when normalized to the lung weight, the deposition rate per gram of lung in humans is much smaller than in rats. In the development of a clearance model, a single-compartment model in the lung was used and a general assumption was made that the clearance of the insoluble and moderately soluble nickel compounds (high temperature (green) NiO and Ni(3)S(2), respectively) depends highly on the volume of retained particles in the lungs. As for the highly soluble nickel compound (NiSO(4). 6H(2)O), the clearance rate coefficient was assumed to depend on the retained particle mass and total alveolar surface. These clearance rate coefficients were extrapolated from the rat data. The retention half-times for high temperature (green) NiO and Ni(3)S(2) particles in humans were found to be much longer than in rats, whereas the retention half-time for NiSO(4). 6H(2)O particles was about the same for both species. The lung burden results in humans for various exposure conditions are predicted and the equivalent exposure concentrations for humans which lead to the same lung burdens found in rats were calculated. PMID- 10464044 TI - Food and Drug Administration proposed testing guidelines for reproduction studies. Revision Committee. FDA Guidelines for Developmental Toxicity and Reproduction, Food and Drug Administration. AB - In the United States, the Food and Drug Administration (FDA) is the agency responsible for ensuring that the direct food additives and color additives used in food are safe for all consumers. In order to determine the safety of these additives for consumption, appropriate information and results from a series of tests must be made available to the agency. In 1982, in an effort to provide guidance to the food industry concerning the appropriate tests for the determination of safety, the FDA issued the Toxicological Principles for the Safety Assessment of Direct Food Additives and Color Additives Used in Foods, commonly referred to as the Redbook. In 1993, based on the expansion of technology and the use of food additives, as well as the refinement of the scientific criteria for establishing safety, the FDA updated its guidelines and issued the draft Redbook II. Since Redbook II was issued, additional refinements have been made in the procedures for the multigeneration reproduction study and for the assessment of effects on male reproduction. The latest proposed guidelines for multigeneration studies are provided here. PMID- 10464045 TI - Food and Drug Administration proposed testing guidelines for developmental toxicity studies. Revision Committee. FDA Guidelines for Developmental Toxicity and Reproduction, Food and Drug Administration. AB - The Food and Drug Administration (FDA) is the agency responsible for ensuring that the direct food additives and color additives used in food in the United States are safe for all consumers. In 1982, in an effort to provide guidance concerning appropriate tests, the FDA issued Toxicological Principles for the Safety Assessment of Direct Food Additives and Color Additives Used in Food, commonly known as the Redbook. The Redbook included detailed guidelines for testing the effects of direct and indirect food and color additives on mothers and their developing fetuses. Based on refinements in safety assessment and risk evaluation as well as expansion of knowledge concerning the metabolism and pharmacokinetics of food and color additives, the need to revise and update the 1982 document became apparent. In 1993, Redbook II in draft form was made available for public comment. Since then, test end points and developmental landmarks have been refined. The latest proposed guidelines for developmental toxicity studies are provided here. PMID- 10464046 TI - Review of mononuclear cell leukemia in F-344 rat bioassays and its significance to human cancer risk: A case study using alkyl phthalates. AB - Elevated incidences of mononuclear cell leukemia (MNCL) have been observed in a number of chronic bioassays in the F-344 rat. As this tumor type is unique to the rat and is only common in the F-344 strain, its significance for human cancer risk is unclear. For this reason, a survey of the published literature was undertaken to assess the occurrence and etiology of MNCL in F-344 rats and to evaluate its potential significance to humans using alkyl phthalate data as an example. It was found that MNCL occurs in untreated, aged F-344 rats at a high and variable rate, it is uncommon in most other rat strains, and its background incidence has increased significantly over time. This complicates retrospective data interpretation. MNCL has not been found in other mammalian species and no histologically comparable tumor is found in humans. In general, a statistically significant increase in frequency of a common tumor in the F-344 rat is an insufficient basis for determining that a chemical presents a carcinogenic hazard to humans, particularly when that tumor is not observed in other species. As one example, the alkyl phthalates constitute one group of substances which has been associated with increased MNCL frequency in the F-344 rat after high dietary doses. In evaluating the significance of this increase in MNCL, an extensive toxicological database for phthalates indicates that toxicological effects occur only at relatively high doses, and tumor development (including MNCL) occurs only after an apparent threshold is exceeded. Phthalates are not genotoxic as a class, further supporting the hypothesis of the existence of a threshold. When these considerations are collectively evaluated, it can be concluded that a finding of increased MNCL in F-344 rats exposed for a lifetime to a nongenotoxic chemical is not toxicologically relevant to humans, even when MNCL is observed at an increased incidence that is statistically significant. Thus, the increased incidence of MNCL observed in F-344 rats exposed to some alkyl phthalates is likely a strain-specific effect of little or no relevance for humans, and characterization of these chemicals as carcinogens based on increased MNCL in F 344 rats is not scientifically supported. PMID- 10464047 TI - Hazard assessment and risk analysis of two new synthetic vitreous fibers. AB - Isofrax and Insulfrax are two new synthetic vitreous fibers (SVFs) developed for high-temperature insulation (1800-2300 degrees F) applications. In an attempt to significantly reduce or eliminate the potential of adverse health effects, these two fibers were specifically designed to have high solubility and, thus, low in vivo biodurability. In this paper, we review the effects of chemical composition on biodurability, in vitro fiber dissolution rates (K(dis)), and the relevance and relationship of K(dis) to pulmonary fibrosis and lung tumors in chronic rat inhalation studies. We also examine the correlations between K(dis) and weighted in vivo half-life (t(0.5)) of long fibers (>20 microm) and their relation to pulmonary effects in chronic rat inhalation bioassays. Predictions for outcomes of inhalation bioassays and development of nonsignificant risk levels of exposure are provided. Additionally, justification for the use of inhalation versus noninhalation animal data is provided as is a brief review of human health effects of SVFs. We conclude, inter alia, that Isofrax and Insulfrax have low biodurability, would not be expected to produce either pulmonary fibrosis or lung tumors in a well-designed animal inhalation bioassay, have weighted half-lives beneath the threshold established by the European Union for classification as a carcinogen, and based on epidemiological data for SVFs would not be expected to result in incremental cancer in human cohorts. Finally, it is estimated that approximately 90% of workplace exposure concentrations of these materials would be beneath 1 f/cc. At a concentration of 1 f/cc, neither fiber would be expected to result in an incremental working lifetime cancer risk greater than 10(-5). PMID- 10464048 TI - Determining modes of action for biologically based risk assessments. PMID- 10464049 TI - Assessment of the stability of N-terminal pro-brain natriuretic peptide in vitro: implications for assessment of left ventricular dysfunction. AB - Plasma concentrations of N-terminal pro-brain natriuretic peptide (NT-proBNP) are raised in patients with left ventricular dysfunction. Measurement of this peptide has a potential diagnostic role in the identification and assessment of patients with heart failure. The stability of this peptide over time periods and conditions pertaining to routine clinical practice has not been reported previously. Blood samples were obtained from 15 subjects. One aliquot was processed immediately, and the remaining portions of the blood samples were stored for 24 h or 48 h at room temperature or on ice prior to processing. Plasma concentrations of NT-proBNP were measured with a novel immunoluminometric assay developed within our laboratory. Mean plasma concentrations of NT-proBNP were not significantly different whether blood samples were centrifuged immediately and stored at -70 degrees C or kept at room temperature or on ice for 24 h or 48 h. The mean percentage differences from baseline (reference standard) were +5.2% (95% confidence interval +18.2 to -7.8%) and +0.8% (+15.2 to -13.7%) after storage for 24 h at room temperature or on ice respectively, and +8.9% (+24.2 to 6. 5%) and +3.2% (+15.1 to -0.9%) for storage for 48 h at room temperature or on ice respectively. Pearson correlation coefficients for baseline NT-proBNP concentrations compared with levels at 48 h at room temperature or on ice were r=0.89 and r=0.83 respectively (both P<0.0001). Thus NT-proBNP extracted from plasma samples treated with EDTA and aprotinin is stable under conditions relevant to clinical practice. PMID- 10464050 TI - Role of cardiac structural and functional abnormalities in the pathogenesis of hyperdynamic circulation and renal sodium retention in cirrhosis. AB - The aim of this study was to assess the relationship between subtle cardiovascular abnormalities and abnormal sodium handling in cirrhosis. A total of 35 biopsy-proven patients with cirrhosis with or without ascites and 14 age matched controls underwent two-dimensional echocardiography and radionuclide angiography for assessment of cardiac volumes, structural changes and systolic and diastolic functions under strict metabolic conditions of a sodium intake of 22 mmol/day. Cardiac output, systemic vascular resistance and pressure/volume relationship (an index of cardiac contractility) were calculated. Eight controls and 14 patients with non-ascitic cirrhosis underwent repeat volume measurements and the pressure/volume relationship was re-evaluated after consuming a diet containing 200 mmol of sodium/day for 7 days. Ascitic cirrhotic patients had significant reductions in (i) cardiac pre-load (end diastolic volume 106+/-9 ml; P<0.05 compared with controls), due to relatively thicker left ventricular wall and septum (P<0.05); (ii) afterload (systemic vascular resistance 992+/-84 dyn.s.cm(-5); P<0. 05 compared with controls) due to systemic arterial vasodilatation; and (iii) reversal of the pressure/volume relationship, indicating contractility dysfunction. Increased cardiac output (6.12+/-0.45 litres/min; P<0.05 compared with controls) was due to a significantly increased heart rate. Pre-ascitic cirrhotic patients had contractile dysfunction, which was accentuated when challenged with a dietary sodium load, associated with renal sodium retention (urinary sodium excretion 162+/-12 mmol/day, compared with 197+/ 12 mmol/day in controls; P<0.05). Cardiac output was maintained, since the pre load was normal or increased, despite a mild degree of ventricular thickening, indicating some diastolic dysfunction. We conclude that: (i) contractile dysfunction is present in cirrhosis and is aggravated by a sodium load; (ii) an increased pre-load in the pre-ascitic patients compensates for the cardiac dysfunction; and (iii) in ascitic patients, a reduced afterload, manifested as systemic arterial vasodilatation, compensates for a reduced pre-load and contractile dysfunction. Cirrhotic cardiomyopathy may well play a pathogenic role in the complications of cirrhosis. PMID- 10464051 TI - Inter-relationships between small, dense low-density lipoprotein (LDL), plasma triacylglycerol and LDL apoprotein B in an atherogenic lipoprotein phenotype in free-living subjects. AB - A predominance of small, dense low-density lipoprotein (LDL) is a major component of an atherogenic lipoprotein phenotype, and a common, but modifiable, source of increased risk for coronary heart disease in the free-living population. While much of the atherogenicity of small, dense LDL is known to arise from its structural properties, the extent to which an increase in the number of small, dense LDL particles (hyper-apoprotein B) contributes to this risk of coronary heart disease is currently unknown. This study reports a method for the recruitment of free-living individuals with an atherogenic lipoprotein phenotype for a fish-oil intervention trial, and critically evaluates the relationship between LDL particle number and the predominance of small, dense LDL. In this group, volunteers were selected through local general practices on the basis of a moderately raised plasma triacylglycerol (triglyceride) level (>1.5 mmol/l) and a low concentration of high-density-lipoprotein cholesterol (<1.1 mmol/l). The screening of LDL subclasses revealed a predominance of small, dense LDL (LDL subclass pattern B) in 62% of the cohort. As expected, subjects with LDL subclass pattern B were characterized by higher plasma triacylglycerol and lower high density lipoprotein cholesterol (<1.1 mmol/l) levels and, less predictably, by lower LDL cholesterol and apoprotein B levels (P<0.05; LDL subclass A compared with subclass B). While hyper-apoprotein B was detected in only five subjects, the relative percentage of small, dense LDL-III in subjects with subclass B showed an inverse relationship with LDL apoprotein B (r=-0.57; P<0.001), identifying a subset of individuals with plasma triacylglycerol above 2.5 mmol/l and a low concentration of LDL almost exclusively in a small and dense form. These findings indicate that a predominance of small, dense LDL and hyper apoprotein B do not always co-exist in free-living groups. Moreover, if coronary risk increases with increasing LDL particle number, these results imply that the risk arising from a predominance of small, dense LDL may actually be reduced in certain cases when plasma triacylglycerol exceeds 2.5 mmol/l. PMID- 10464052 TI - Forearm vasoconstriction in response to noradrenaline and NG-monomethyl-L arginine in essential hypertension. AB - A role for abnormal NO production in essential hypertension remains controversial. Blunted vasoconstriction of forearm resistance vasculature in response to N(G)-monomethyl-L-arginine (L-NMMA; an inhibitor of NO biosynthesis), relative to the response to noradrenaline, has been reported in hypertensive patients and interpreted as evidence of reduced basal NO biosynthesis. We sought to determine whether reduced sensitivity of forearm vasculature to the vasoconstrictor action of L-NMMA relative to that of noradrenaline is a consistent finding in essential hypertension. We studied a group of patients (n=32; blood pressure 176+/-4/102+/-2 mmHg; means+/-S.E.M.) and a group of healthy normotensive controls (n=32; blood pressure 130+/-2/75+/-1 mmHg). Noradrenaline (60-240 pmol.min(-1)) and L-NMMA (1-4 micromol.min(-1)) were infused into the brachial artery, and forearm blood flow was measured by venous occlusion plethysmography. The effects of each vasoconstrictor were similar in hypertensive and control subjects. The highest dose of L-NMMA reduced forearm blood flow by 0.75+/-0.12 ml.min(-1).dl(-1) in the control group and by 0.89+/ 0.10 ml.min(-1).dl(-1) in the hypertensive group. The study had 90% power (with P=0.05) to detect a 10% difference in forearm blood flow response between the hypertensive and control groups. We conclude that reduced sensitivity of forearm resistance vasculature to the vasoconstrictor action of L-NMMA is not a universal feature of essential hypertension. This argues against a primary role for reduced basal NO biosynthesis in skeletal muscle resistance vessels in the pathogenesis of essential hypertension. PMID- 10464053 TI - Lack of effect of clinical doses of cyclosporin A on erythrocyte Na+/K+-ATPase activity. AB - Cyclosporin A (CsA) may exert its cytotoxic effects by altering the activity of different plasma membrane transport systems. Although CsA may act at the gene level, it has been also suggested that it can directly alter transport processes at the plasma membrane. To examine this possibility in a physiological context, we determined Na(+)/K(+)-ATPase activity in erythrocytes from two groups of subjects receiving CsA treatment: group I consisted of kidney transplant patients, and group II comprised patients with steroid-resistant idiopathic nephrotic syndrome. Group I patients showed a marked decrease (35%) in the activity of the Na(+)/K(+)-ATPase in erythrocytes immediately after surgery, before the initiation of CsA treatment. The activity remained low 2 days after the introduction of CsA, but had recovered to the original (pre-surgery) value 1 month later. Group II patients showed the same pattern of erythrocyte Na(+)/K(+) ATPase activity as those in group I. When the blood CsA levels from all patients were plotted against the corresponding erythrocyte Na(+)/K(+)-ATPase transport activity, a significant linear correlation was found. Higher levels of CsA in the blood were correlated significantly with increased Na(+)/K(+)-ATPase activities. The blood sodium concentration was also correlated positively with both erythrocyte Na(+)/K(+)-ATPase activity and blood CsA concentration. Thus CsA treatment is not associated with inhibition of the Na(+)/K(+)-ATPase in erythrocytes. PMID- 10464054 TI - Continuous stroke volume monitoring by modelling flow from non-invasive measurement of arterial pressure in humans under orthostatic stress. AB - The relationship between aortic flow and pressure is described by a three-element model of the arterial input impedance, including continuous correction for variations in the diameter and the compliance of the aorta (Modelflow). We computed the aortic flow from arterial pressure by this model, and evaluated whether, under orthostatic stress, flow may be derived from both an invasive and a non-invasive determination of arterial pressure. In 10 young adults, Modelflow stroke volume (MFSV) was computed from both intra-brachial arterial pressure (IAP) and non-invasive finger pressure (FINAP) measurements. For comparison, a computer-controlled series of four thermodilution estimates (thermodilution determined stroke volume; TDSV) were averaged for the following positions: supine, standing, head-down tilt at 20 degrees (HDT20) and head-up tilt at 30 degrees and 70 degrees (HUT30 and HUT70 respectively). Data from one subject were discarded due to malfunctioning thermodilution injections. A total of 155 recordings from 160 series were available for comparison. The supine TDSV of 113+/-13 ml (mean+/-S.D.) dropped by 40% to 68+/-14 ml during standing, by 24% to 86+/-12 ml during HUT30, and by 51% to 55+/-15 ml during HUT70. During HDT20, TDSV was 114+/-13 ml. MFSV for IAP underestimated TDSV during HDT20 (-6+/-6 ml; P<0.05), but that for FINAP did not (-4+/-7 ml; not significant). For HUT70 and standing, MFSV for IAP overestimated TDSV by 11+/-10 ml (HUT70; P<0.01) and 12+/ 9 ml (standing; P<0.01). However, the offset of MFSV for FINAP was not significant for either HUT70 (3+/-8 ml) or standing (3+/-9 ml). In conclusion, due to orthostasis, changes in the aortic transmural pressure may lead to an offset in MFSV from IAP. However, Modelflow correctly calculated aortic flow from non-invasively determined finger pressure during orthostasis. PMID- 10464055 TI - Heterogeneous vascular endothelial growth factor (VEGF) isoform mRNA and receptor mRNA expression in human glomeruli, and the identification of VEGF148 mRNA, a novel truncated splice variant. AB - Vascular endothelial growth factor (VEGF) mediates increased vascular permeability and endothelial mitogenesis, and may orchestrate normal glomerular permselectivity and proteinuria. Distinct isoforms result from differential gene splicing. VEGF binds to two cell surface tyrosine-kinase receptors, KDR (kinase domain region) and Flt-1 (fms-like tyrosine kinase-1). The latter also exists in a soluble form (sFlt), which is inhibitory. We have studied patterns of VEGF isoform and VEGF-receptor expression in isolated single normal human glomeruli. mRNA from 190 glomeruli (from 20 individuals) was harvested on to magnetic beads, and nested reverse transcription-PCR was performed using primers for the VEGF isoforms and VEGF receptors. Simultaneous nested reverse transcription-PCR for CD45 was conducted in order to exclude leucocyte contamination. Unexpected products were isolated, cloned and sequenced. Multiple patterns of glomerular VEGF mRNA isoform expression were identified. Most frequently (58%), all three common forms were expressed. VEGF(189) (i.e. 189-amino-acid form of VEGF) was expressed in 63%, VEGF(165) in 85% and VEGF(121) in 84% of glomeruli. Two unexpected PCR products were also identified: 18% of glomeruli expressed VEGF(145), and 27% of glomeruli expressed a new truncated VEGF splice variant, VEGF(148), lacking exon 6, the terminal part of exon 7 and exon 8. Multiple patterns of VEGF-receptor expression were also identified, the most common being expression of all three isoforms (28%). Overall, KDR was seen in 59% of glomeruli, Flt-1 in 45% and sFlt in 57%. Thus the expression of VEGF within normal glomeruli is complex and variable, with inter- and intra-individual variation. Furthermore, sFlt appears to be the co-dominant form of VEGF receptor expressed within glomeruli, suggesting that, in healthy individuals, a degree of VEGF autoregulation is the norm. The physiological importance of VEGF(148) remains to be confirmed. PMID- 10464056 TI - Relationship between vascular reactivity in vitro and blood flows in rats with cirrhosis. AB - In cirrhosis there is a hyperdynamic circulation, which occurs mainly in the systemic and splanchnic regions. Using isolated-vessel models, previous studies have shown reduced aortic reactivity to vasoconstrictors in rats with cirrhosis. The aim of the present study was to evaluate and compare the vascular responsiveness to phenylephrine in arterial rings and the blood flows from different regions in rats with cirrhosis and controls. Reactivity was studied in isolated thoracic aortic, superior mesenteric arterial and carotid arterial rings from sham-operated and bile-duct-ligated rats by measuring the cumulative concentration-dependent tension induced by phenylephrine (10(-9)-10(-4) M). Blood flows were measured by the radioactive microsphere method. In rats with cirrhosis, a significant hyporeactivity to phenylephrine was observed in both the aorta and the superior mesenteric artery compared with the corresponding arteries of normal rats. This hyporesponsiveness was corrected by N(omega)-nitro-L arginine (0.1 mM). In contrast, carotid artery reactivity and the responses to N(omega)-nitro-L-arginine were similar in the cirrhotic and control groups. In each case, cardiac output and mesenteric arterial blood flow were significantly higher in cirrhotic than in normal rats. Cerebral blood flows were not significantly different between the two groups. In cirrhotic rats, arterial hyporeactivity may be a consequence of increased regional blood flow and increased production of nitric oxide. PMID- 10464057 TI - Effect of aging on CD11b and CD69 surface expression by vesicular insertion in human polymorphonuclear leucocytes. AB - The exocytosis of intracellular vesicles is an important function of the plasma membrane, which is responsible for hormone secretion, cell surface expression of antigens, ion transporters and receptors, and intracellular and intercellular signalling. Human aging is associated with many physiological and cellular changes, many of which are due to alterations in plasma membrane functioning. Alterations in vesicle externalization with age could account for many of these changes. We investigated whether alterations in vesicle exocytosis occur with increasing age by flow-cytometric determination of CD11b and CD69 expression on the surface of human polymorphonuclear leucocytes (PMN) stimulated with phorbol myristate acetate (PMA), a tumour promoter which binds to and activates protein kinase C (PKC) directly, or with formyl-Met-Leu-Phe (fMLP), which activates PKC indirectly via interactions with a cell surface receptor and G-protein, and subsequent inositol phosphate hydrolysis. Following stimulation with PMA, a decrease in the proportion of PMN expressing CD69 at high levels was observed in elderly compared with young subjects (young, 55.3%; elderly, 43.9%; P=0.01). No aging-related differences in the proportion of PMN expressing CD11b (young, 73.7%; elderly, 68.4%; P=0.15), or in the number of molecules of CD69 or CD11b expressed per cell, were observed. Stimulation with fMLP or low PMA concentrations resulted in full CD11b expression but minimal CD69 expression in both young and elderly subjects. Cells which expressed CD69 had no CD11b expression, while those cells expressing CD11b had minimal CD69 expression. Thus the PMA-induced expression of CD11b and CD69 in human PMN represents two separate processes, only one of which is affected in aging. CD11b expression appears to require a lesser degree of PKC stimulation compared with that required for CD69 expression. The age-associated reduction in PMA-stimulated CD69 expression may occur either at or distal to PKC activation. Such a decrease may contribute to the age-associated impairments in PMN function that contribute, in turn, to immunosenescence. PMID- 10464058 TI - Stress response decreases the interleukin-1beta-induced production of complement component C3 in human intestinal epithelial cells. AB - Interleukin-1beta (IL-1beta) increases the production of complement component C3 in enterocytes. Heat shock regulates the response to cytokines and other inflammatory mediators in various cell types. We tested the hypothesis that the heat-shock response regulates IL-1beta-induced C3 production in the enterocyte. Cultured Caco-2 cells, a human intestinal epithelial cell line, were treated with sodium arsenite (10-500 microM) for 1 h or subjected to hyperthermia (43 degrees C) for 1-4 h, and allowed to recover for 1 h. The cells were then treated with IL 1beta (0.5 ng/ml) for up to 24 h, whereafter C3 levels were measured by ELISA and C3 mRNA by Northern blot analysis. Heat-shock protein of 72 kDa (hsp72) was determined by Western blot analysis. Treatment of the cells with sodium arsenite or subjecting them to hyperthermia induced the expression of hsp72. The IL-1beta induced expression of C3 mRNA and C3 production were down-regulated by hyperthermia and sodium arsenite in a dose-dependent fashion. The results suggest that the stress response induced by hyperthermia or sodium arsenite decreases IL 1beta-induced C3 production in human enterocytes. PMID- 10464059 TI - Nitric oxide in human hypertension-up, down or unaffected? PMID- 10464060 TI - Neither endogenous nor inhaled nitric oxide influences the function of circulating platelets in healthy volunteers. AB - Experimental models have indicated prothrombotic effects of inhibition of nitric oxide (NO) production, and anti-thrombotic effects of inhaled NO, but the influence of NO on platelet function in vivo in humans is not well established. We therefore investigated the effects of systemic inhibition of NO synthesis by N(G)-monomethyl-L-arginine (L-NMMA) and of NO inhalation on platelet function in vivo. On two occasions, L-NMMA (13.5 mg/kg) or saline infusion was administered to 14 healthy volunteers in a double-blind cross-over study. After a 30 min infusion of L-NMMA or placebo, NO inhalation (30 p.p.m) was added during the remaining 30 min of infusion, on both occasions. Measurements included filtragometry ex vivo (reflecting platelet aggregability), flow-cytometric evaluation of platelets in whole blood (fibrinogen binding and P-selectin expression), plasma beta-thromboglobulin (reflecting platelet secretion), cGMP in platelets and plasma, thrombin generation markers (thrombin fragment 1+2 and thrombin-antithrombin complexes) in plasma, and bleeding time. L-NMMA increased blood pressure and decreased heart rate. NO inhalation did not influence blood pressure or heart rate, but caused a 3-fold elevation in plasma cGMP levels (P<0.001). Neither L-NMMA nor NO influenced filtragometry readings or flow cytometric determinations of platelet fibrinogen binding and P-selectin expression. Furthermore, plasma beta-thromboglobulin, platelet cGMP and thrombin generation markers were not influenced by either treatment. Bleeding time was not influenced by L-NMMA compared with placebo, but was increased by approximately 25% during NO inhalation (P<0.01), whether NO synthesis had been inhibited or not. The prolongation of bleeding time by inhaled NO was not accompanied by any effect on the platelet variables assessed. The present results indicate that circulating platelets are not influenced by endogenous or inhaled NO, presumably due to the rapid inactivation of NO in the blood. This does not exclude possible effects of endothelial NO in the interface between the blood and the vessel wall. PMID- 10464061 TI - Tissue- and isoform-specific effects of aging in rats on protein kinase C in insulin-sensitive tissues. AB - The mechanisms responsible for the age-related decline in insulin sensitivity have not been clearly identified, but activation of the diacylglycerol/protein kinase C (PKC) signalling pathway (often confined to individual isoforms of PKC) has recently been implicated in the pathogenesis of other insulin-resistant states in both humans and animal models. Fasting serum glucose, insulin and triacylglycerol (triglyceride) concentrations, and results of oral glucose tolerance tests, were compared in groups of 6-week-old (n=8) and 6-month-old (n=8) Sprague-Dawley rats. Insulin-responsive tissues (liver, soleus muscle and epididymal fat pad) were collected to compare levels of diacylglycerol, PKC enzyme activity and protein expression of individual PKC isoforms in cytosol and membrane fractions. The older group were heavier (556+/-14 g, compared with 188+/ 7 g) and relatively insulin-resistant and hyperinsulinaemic (477+/-73 pM compared with 293+/-51 pM; P<0.05) compared with young rats; they also had greater areas under the serum glucose (old, 20. 3+/-1.1; young, 17.3+/-0.7 mmol.h(-1).l(-1)) and insulin (old, 1254+/-76; young, 721+/-113 mmol.h(-1).l(-1)) profiles following an oral glucose tolerance test, and significantly higher fasting triacylglycerol levels (old, 1.24+/-0.06 mM; young, 0.92+/-0.07 mM; P<0.01). There were no age-related differences in diacylglycerol levels or PKC activity in muscle and liver, but membrane-associated PKC activity was 2.5-fold higher in the adipose tissue of older rats (101+/-19 compared with 40+/-5 pmol.min(-1).mg(-1) protein; P<0.05) due to increased translocation of PKC-beta(I), -beta(II) and epsilon. Thus insulin resistance due to normal aging is associated with tissue- and isoform-specific changes in diacylglycerol/PKC signalling. In contrast with diabetes and dietary-induced insulin resistance, there were no changes in diacylglycerol/PKC signalling in skeletal muscle and liver, but isoform-specific translocation and higher PKC activity in adipose tissue may blunt the insulin mediated inhibition of lipolysis and contribute to the increased triacylglycerol levels observed in older animals. PMID- 10464062 TI - Effects of acidosis on leptin secretion from 3T3-L1 adipocytes and on serum leptin in the uraemic rat. AB - Marked hyperleptinaemia and metabolic acidosis are common findings in patients with chronic renal failure. In animal models, both leptin administration and acidosis reduce food intake. However, leptin causes loss of body fat, while acidosis induces muscle wasting. Whether a low pH and leptin production are related has not been studied. Leptin secretion was measured in cultured 3T3-L1 adipocytes exposed to acid or control pH for up to 96 h. In addition, serum leptin was compared between acidotic and bicarbonate-treated uraemic Wistar rats using the remnant model. Leptin levels in the culture medium were decreased at an acid pH of 7.1 compared with a control pH of 7.5 at 96 h (562+/-78 and 831+/-103 pg.48 h(-1). well(-1) respectively; mean+/-S.E.M.; P=0.037). Similarly, serum leptin in uraemic rats was found to be lower in the acidotic group than in the bicarbonate-treated group, although this observation fell just short of statistical significance (1273+/-171 compared with 2059+/-376 pg/ml; P=0.07). In conclusion, acidosis decreases leptin secretion from cultured adipocytes. Accordingly, acidotic uraemic rats seem to exhibit lower serum leptin levels than their bicarbonate-supplemented counterparts. This study is the first report providing a link between acidosis and leptin levels. PMID- 10464063 TI - Decreased activity of the red blood cell ATPase-dependent Na+ pump in patients with cardiac syndrome X. AB - Marked Na(+)/Li(+) countertransport hyperactivity and post-load hyperinsulinaemia have been described in 93% of patients with cardiac syndrome X. We hypothesized that more complex abnormalities in Na(+) traffic across the cell membrane are present in these patients. The aim of the present study was to evaluate the activities of the two main transporters responsible for transmembrane Na(+) transport, i.e. the ATPase-dependent Na(+) pump and the Na(+)-K(+)-2Cl(-) co transporter, in a selected group of patients with cardiac syndrome X. We evaluated 19 patients with cardiac syndrome X and 14 control subjects. The ATPase dependent Na(+) pump and Na(+)-K(+)-2Cl(-) co-transport activities were assessed from Na(+)-loaded red blood cells by using nystatine, in the presence of furosemide and ouabain, as appropriate. Erythrocyte Na(+)/Li(+) countertransport activity, serum lipid and post-load (75 g of oral glucose) insulin levels were also evaluated. The V(max) of Na(+)/Li(+) countertransport (P=0.0001) and post load insulin levels (120 min; P=0.001) were confirmed to be higher in patients with syndrome X than in controls. The V(max) of Na(+)-K(+)-2Cl(-) co-transport was similar in patients and controls. By contrast, the V(max) of the ATPase dependent Na(+) pump was significantly lower (P=0.002) in syndrome X patients (3.13+/-0.87 mmol.h(-1).l(-1)) than in controls (4.28+/-1.10 mmol.h(-1).l(-1)). Serum total cholesterol and triacylglycerol concentrations were also higher in patients with syndrome X than in control subjects (P<0.0001). Thus decreased activity of the ATPase-dependent Na(+) pump was present in patients with cardiac syndrome X. Such an abnormality has the biological potential to augment microvascular tone and the response to constrictor stimuli via increased intracellular free Ca(2+). Of note, syndrome X patients also manifested Na(+)/Li(+) countertransport hyperactivity which, in turn, is known to induce peripheral insulin resistance and consequent abnormalities in insulin secretion and lipid turnover. Thus cardiac syndrome X appears as a multifaceted syndrome presenting with either metabolic or cardiovascular symptoms, or both, based on the expression of complex abnormalities in Na(+) traffic across the cell membrane. PMID- 10464064 TI - Discrepancies between echocardiographic measurements of left ventricular mass in a healthy adult population. AB - Increased left ventricular (LV) mass is associated with increased cardiovascular morbidity and mortality. LV mass is commonly estimated from echocardiography according to the Penn or ASE (American Society of Echocardiography) conventions. No formal statistical test of agreement between these methods has been published. Therefore we compared M-mode echocardiographic LV mass estimates by the Penn and ASE methods in a normal adult population. M-mode echocardiographic tracings were obtained in 169 healthy volunteers and used to calculate LV mass using the Penn and ASE methods. Median values of the estimates were similar [Penn, 126 g (interquartile range 96-170 g); ASE, 129 g (105-164 g); P=0.08] and were highly intercorrelated (r=0.98, P<0.0001). However, the Bland-Altman analysis of agreement revealed significant inconsistencies between Penn and ASE LV mass values. The difference between Penn and ASE values was correlated significantly with heart size (P<0.0001), such that, for small hearts, the Penn LV mass was lower than the ASE LV mass; in contrast, for large hearts, Penn estimates were greater than ASE values. In the upper 5% of the LV mass distribution, the median value for the Penn LV mass index was 132.4 g/m(2), compared with 116.5 g/m(2) for ASE values (2P=0.017). Thus the two most common methods of echocardiographic estimation of LV mass differ significantly at the upper and lower ends of the heart size distribution. These results have important implications for both cardiac research and clinical evaluation. PMID- 10464066 TI - Air pollution and bronchitic symptoms in Southern California children with asthma. AB - People who live in cities with dirty air have blacker lungs than people who live in rural areas with less air pollution. This is because, although particulates larger than 10 microm are filtered out when inhaled air passes through the nose, smaller particulates reach the lower airways. The particulates that reach the alveoli (the terminal air pockets of the lungs) stay there permanently. This accounts for the fact that a person who has lived in a polluted city for many years has blacker lungs than one who has lived in a polluted city for a shorter time. PMID- 10464065 TI - Interleukin-8 stimulates the migration of human colonic epithelial cells in vitro. AB - The migration of colonic epithelial cells (restitution) is an important event in the repair of mucosal injuries. Interleukin-8 (IL-8) is a physiological initiator of the chemotactic migration of leucocytes. This study aimed to determine whether IL-8 had a similar effect on migration in an in vitro model of wounded colonic epithelium. Cell migration over 24 h was assessed in circular wounds made in confluent monolayers of the human colon cancer cell line LIM1215. This migration was stimulated in a concentration-dependent manner by IL-8, with maximal effects of approx. 1.75-fold above basal migration. The motogenic effect of IL-8 was mediated independently of effects on cell proliferation. In contrast, it was partially dependent upon gene transcription and protein synthesis and involved the activation of pertussis-toxin-sensitive G-proteins. The short-chain fatty acids, acetate, propionate, butyrate and valerate, the activator of protein kinase C (phorbol-12-myristate-13-acetate) and tumour necrosis factor-alpha (TNF alpha) all stimulated the secretion of IL-8. However, only the motogenic effect of TNF-alpha was dependent upon IL-8. In conclusion, IL-8 stimulated cell migration in an in vitro model of colonic epithelium, whereas the motogenic effect of at least one physiologically relevant factor was dependent upon an increase in its endogenous levels. If IL-8 stimulates colonic epithelial restitution in vivo, this would have ramifications for the control of repair processes following wounding of the colonic mucosa. PMID- 10464067 TI - Meta-analysis of rat lung tumors from lifetime inhalation of diesel exhaust. AB - Estimating the carcinogenic potential of exposure to diesel-engine exhaust particulates (DEPs) is problematic. In rats, high concentrations of DEPs (> 1,000 microg/m(3)) inhaled over a lifetime result in excess lung tumors. However, data for rats exposed to DEP at concentrations not associated with lung overload are consistent with no tumorigenic effect. Individual rat studies have only a limited number of exposure groups; therefore, we combined the tumor data from eight chronic inhalation studies in a meta-analysis. Statistical analysis identified a threshold of response between 200 and 600 microg/m(3) average continuous lifetime exposure, consistent with biological-effect thresholds reported by other investigators. Our exposure-response analysis of all rats with < 600 microg/m(3) average continuous lifetime exposure found no tumorigenic effect of DEP in these rats. When we evaluated all rat studies, accounted for a threshold and for inhomogeneity between experiments, and expressed the results in terms of human unit risk (UR), we found a negative maximum-likelihood human UR of -32 (times) 10(-6) per microgram per cubic meter (microg/m(3)), but this was not statistically significantly different from zero. Extrapolating the rat upper 95th percentile confidence limit to humans gave an upper-bound human UR of 9.3 (times) 10(-6) per microg/m(3)]. This upper-bound human UR, derived from all of the data points (including 1,087 animals below the estimated threshold and 1,433 in the control groups), falls entirely below the range of estimates derived from lung overloaded rats or from epidemiology of railroad workers. Our meta-analysis of the low-exposure data in rats does not support a lung cancer risk for DEP exposure at nonoverload conditions. Average ambient concentrations of DEP (0-3 microg/m(3)) are < 1% of the concentration associated here with a threshold of tumor response in the rat bioassay. PMID- 10464068 TI - Quantification of two aromatic amine mutagens, PBTA-1 and PBTA-2, in the yodo river system. AB - The levels of two aromatic amine mutagens, 2-[2-(acetylamino)-4-[bis(2 methoxyethyl)amino]-5-methoxyphenyl]-5-am ino-7-bromo-4-chloro-2H-benzotriazole (PBTA-1) and 2-[2-(acetylamino-4-[N-(2-cyanoethyl)ethylamino]-5-methoxyphenyl]-5 a mino-7-bromo-4-chloro-2H-benzotriazole (PBTA-2), were quantitatively analyzed in the Yodo River system in Japan. The river water samples were collected at nine sampling sites from the Yodo River system twice or three times between May and July in 1997. PBTA-1 and PBTA-2 in the river water samples were concentrated on blue rayon columns, partially purified by high-performance liquid chromatography (HPLC) on reverse-phase columns, then quantified by HPLC with an electrochemical detector. The amounts of PBTA-1 and PBTA-2 in the water samples were < 0.01-1.91 and < 0.01-2.25 ng/L, respectively. High levels of PBTA-1 and PBTA-2 were detected in the samples collected within 4 km downstream of two sewage plants, which are located along the banks of the Nishitakase River, a tributary of the Yodo River system, and these samples showed stronger mutagenicity in Salmonella typhimurium YG1024 with S9 mix than the other water samples. On the other hand, the river water samples from upstream of the sewage plant were weakly or not mutagenic and PBTA-1 and PBTA-2 were not detected. These results confirmed that a major source of PBTA-1 and PBTA-2 in the Yodo River system is effluent from the sewage plants and that discharged mutagens, including PBTA-1 and PBTA-2, are diluted and/or decomposed while moving down the Yodo River system. PMID- 10464070 TI - Reporter gene assay for fish-killing activity produced by Pfiesteria piscicida. AB - Collaborative studies were performed to develop a functional assay for fish killing activity produced by Pfiesteria piscicida. Eight cell lines were used to screen organic fractions and residual water fraction by using a 3-[4, 5 dimethylthiazol-(2-4)]-diphenyltetrazolium bromide cytotoxicity assay. Diethyl ether and a residual water fraction were cytotoxic to several cell lines including rat pituitary (GH(4)C(1)) cells. Residual water as well as preextracted culture water containing P. piscicida cells induced c-fos-luciferase expressed in GH(4)C(1) cells with a rapid time course of induction and sensitive detection. The reporter gene assay detected activity in toxic isolates of P. piscicida from several North Carolina estuaries in 1997 and 1998 and may also be suitable for detecting toxic activity in human and animal serum. PMID- 10464069 TI - Arsenic concentrations in well water and risk of bladder and kidney cancer in Finland. AB - We assessed the levels of arsenic in drilled wells in Finland and studied the association of arsenic exposure with the risk of bladder and kidney cancers. The study persons were selected from a register-based cohort of all Finns who had lived at an address outside the municipal drinking-water system during 1967-1980 (n = 144,627). The final study population consisted of 61 bladder cancer cases and 49 kidney cancer cases diagnosed between 1981 and 1995, as well as an age- and sex-balanced random sample of 275 subjects (reference cohort). Water samples were obtained from the wells used by the study population at least during 1967 1980. The total arsenic concentrations in the wells of the reference cohort were low (median = 0.1 microg/L; maximum = 64 microg/L), and 1% exceeded 10 microg/L. Arsenic exposure was estimated as arsenic concentration in the well, daily dose, and cumulative dose of arsenic. None of the exposure indicators was statistically significantly associated with the risk of kidney cancer. Bladder cancer tended to be associated with arsenic concentration and daily dose during the third to ninth years prior to the cancer diagnosis; the risk ratios for arsenic concentration categories 0.1-0.5 and [Greater/equal to] 0.5 microg/L relative to the category with < 0.1 microg/L were 1.53 [95% confidence interval (CI), 0.75-3.09] and 2.44 (CI, 1.11-5.37), respectively. In spite of very low exposure levels, we found some evidence of an association between arsenic and bladder cancer risk. More studies are needed to confirm the possible association between arsenic and bladder cancer risk at such low exposure levels. PMID- 10464071 TI - Chloracne, goiter, arthritis, and anemia after polychlorinated biphenyl poisoning: 14-year follow-Up of the Taiwan Yucheng cohort. AB - In 1979, a mass poisoning involving 2,000 people occurred in central Taiwan from ingestion of cooking oil contaminated by polychlorinated biphenyls (PCBs) and polychlorinated dibenzofurans (PCDFs). We studied the prevalence of medical conditions in the exposed individuals and in a neighborhood control group. Starting with a registry of the exposed individuals from 1983, we updated the addresses of exposed individuals and identified a control group matched for age, sex, and neighborhood in 1979. In 1993, individuals 30 years of age or older were interviewed by telephone. We obtained usable information from 795 exposed subjects and 693 control subjects. Lifetime prevalence of chloracne, abnormal nails, hyperkeratosis, skin allergy, goiter, headache, gum pigmentation, and broken teeth were observed more frequently in the PCB/PCDF-exposed men and women. The exposed women reported anemia 2. 3 times more frequently than controls. The exposed men reported arthritis and herniated intervertebral disks 4.1 and 2.9 times, respectively, more frequently than controls. There was no difference in reported prevalences of other medical conditions. We conclude that Taiwanese people exposed to high levels of PCBs and PCDFs reported more frequent medical problems, including skin diseases, goiter, anemia, and joint and spine diseases. PMID- 10464072 TI - Distribution of pesticides and polycyclic aromatic hydrocarbons in house dust as a function of particle size. AB - House dust is a repository for environmental pollutants that may accumulate indoors from both internal and external sources over long periods of time. Dust and tracked-in soil accumulate most efficiently in carpets, and the pollutants associated with dust and soil may present an exposure risk to infants and toddlers, who spend significant portions of their time in contact with or in close proximity to the floor and who engage in frequent mouthing activities. The availability of carpet dust for exposure by transfer to the skin or by suspension into the air depends on particle size. In this study, a large sample of residential house dust was obtained from a commercial cleaning service whose clients were homeowners residing in the Raleigh-Durham-Chapel Hill (Research Triangle) area of North Carolina. The composite dust was separated into seven size fractions ranging from < 4 to 500 microm in diameter, and each fraction was analyzed for 28 pesticides and 10 polycyclic aromatic hydrocarbons (PAHs). Over 20% of the fractionated dust sample consisted of particles < 25 microm in diameter. Fourteen pesticides and all 10 of the target PAHs were detected in one or more of the seven size-fractionated samples. Sample concentrations reported range from 0.02 to 22 microg/g; the synthetic pyrethroids cis- and trans permethrin were the most abundant pesticide residue. The concentrations of nearly all of the target analytes increased gradually with decreasing particle size for the larger particles, then increased dramatically for the two smallest particle sizes (4-25 microm and < 4 microm). PMID- 10464073 TI - The relationship of arsenic levels in drinking water and the prevalence rate of skin lesions in Bangladesh. AB - To determine the relationship of arsenic-associated skin lesions and degree of arsenic exposure, a cross-sectional study was conducted in Bangladesh, where a large part of the population is exposed through drinking water. Four villages in Bangladesh were identified as mainly dependent on wells contaminated with arsenic. We interviewed and examined 1,481 subjects [Greater/equal to] 30 years of age in these villages. A total of 430 subjects had skin lesions (keratosis, hyperpigmentation, or hypopigmentation). Individual exposure assessment could only be estimated by present levels and in terms of a dose index, i.e., arsenic levels divided by individual body weight. Arsenic water concentrations ranged from 10 to 2,040 microg/L, and the crude overall prevalence rate for skin lesions was 29/100. After age adjustment to the world population the prevalence rate was 30. 1/100 and 26.5/100 for males and females, respectively. There was a significant trend for the prevalence rate both in relation to exposure levels and to dose index (p < 0.05), regardless of sex. This study shows a higher prevalence rate of arsenic skin lesions in males than females, with clear dose-response relationship. The overall high prevalence rate in the studied villages is an alarming sign of arsenic exposure and requires an urgent remedy. PMID- 10464074 TI - Chronic beryllium disease and cancer risk estimates with uncertainty for beryllium released to the air from the Rocky Flats Plant. AB - Beryllium was released into the air from routine operations and three accidental fires at the Rocky Flats Plant (RFP) in Colorado from 1958 to 1989. We evaluated environmental monitoring data and developed estimates of airborne concentrations and their uncertainties and calculated lifetime cancer risks and risks of chronic beryllium disease to hypothetical receptors. This article discusses exposure response relationships for lung cancer and chronic beryllium disease. We assigned a distribution to cancer slope factor values based on the relative risk estimates from an occupational epidemiologic study used by the U.S. Environmental Protection Agency (EPA) to determine the slope factors. We used the regional atmospheric transport code for Hanford emission tracking atmospheric transport model for exposure calculations because it is particularly well suited for long term annual-average dispersion estimates and it incorporates spatially varying meteorologic and environmental parameters. We accounted for model prediction uncertainty by using several multiplicative stochastic correction factors that accounted for uncertainty in the dispersion estimate, the meteorology, deposition, and plume depletion. We used Monte Carlo techniques to propagate model prediction uncertainty through to the final risk calculations. We developed nine exposure scenarios of hypothetical but typical residents of the RFP area to consider the lifestyle, time spent outdoors, location, age, and sex of people who may have been exposed. We determined geometric mean incremental lifetime cancer incidence risk estimates for beryllium inhalation for each scenario. The risk estimates were < 10(-6). Predicted air concentrations were well below the current reference concentration derived by the EPA for beryllium sensitization. PMID- 10464075 TI - Oil shale processing as a source of aquatic pollution: monitoring of the biologic effects in caged and feral freshwater fish. AB - The biologic effects of the oil shale industry on caged rainbow trout (Oncorhynchus mykiss) as well as on feral perch (Perca fluviatilis) and roach (Rutilus rutilus) were studied in the River Narva in northeast Estonia. The River Narva passes the oil shale mining and processing area and thus receives elevated amounts of polycyclic aromatic hydrocarbons (PAHs), heavy metals, and sulfates. The effects of the chemical load were monitored by measuring cytochrome P4501A (CYP1A)-dependent monooxygenase (MO) activities [7-ethoxyresorufin O-deethylase and aryl hydrocarbon hydroxylase (AHH)] as well as conjugation enzyme activities [glutathione S-transferase (GST) and UDP-glucuronosyltransferase] in the liver of fish. CYP1A induction was further studied by detecting the amount and occurrence of the CYP1A protein. Histopathology of tissues (liver, kidney, spleen, and intestine) and the percentage of micronuclei in fish erythrocytes were also determined. Selected PAHs and heavy metals (Cd, Cu, Hg, and Pb) were measured from fish muscle and liver. In spite of the significant accumulation of PAHs, there was no induction of MO activities in any studied fish species. When compared to reference samples, AHH activities were even decreased in feral fish at some of the exposed sites. Detection of CYP1A protein content and the distribution of the CYP1A enzyme by immunohistochemistry also did not show extensive CYP1A induction. Instead, GST activities were significantly increased at exposed sites. Detection of histopathology did not reveal major changes in the morphology of tissues. The micronucleus test also did not show any evidence of genotoxicity. Thus, from the parameters studied, GST activity was most affected. The lack of catalytic CYP1A induction in spite of the heavy loading of PAHs was not studied but has been attributed to the elevated content of other compounds such as heavy metals, some of which can act as inhibitors for MOs. Another possible explanation of this lack of induction is that through adaptation processes the fish could have lost some of their sensitivity to PAHs. Either complex pollution caused by oil shale processing masked part of the harmful effects measured in this study, or oil shale industry did not have any severe effects on fish in the River Narva. Our study illustrates the difficulties in estimating risk in cases where there are numerous various contaminants affecting the biota. PMID- 10464076 TI - Tumor promoters in commercial indoor-plant cultivars of the Euphorbiaceae. AB - Certain decorative indoor-plant cultivars are derived from toxic wild plant species. Native members of the Euphorbiaceae (spurge) contain highly irritating and tumor-promoting diterpene esters. Plant breeders and gardeners are constantly searching for less toxic cultivars of the popular Euphorbiaceae indoor plants. In this investigation, 22 commercial cultivars of Euphorbiaceae indoor plants were examined for tumor promoter contents by high-performance liquid chromatography (HPLC). Cultivars of E. milii (E. lomii hybrids), and in particular E. leuconeura, contained ingenol derivatives, whereas cultivars of E. pulcherrima and Codiaeum variegatum were devoid of these compounds. Tumor-promoting activity was assessed by induction of a luciferase reporter gene, which was placed under the control of an Epstein-Barr virus early antigen promoter. The response was closely correlated with ingenol ester content; the latex of the two E. leuconeura cultivars tested gave the strongest response. The HPLC and bioassay methods used in this study provide a basis for the development of nontoxic indoor-plant cultivars and perhaps for consumer-oriented labeling. PMID- 10464077 TI - Air pollution and bronchitic symptoms in Southern California children with asthma. AB - The association of air pollution with the prevalence of chronic lower respiratory tract symptoms among children with a history of asthma or related symptoms was examined in a cross-sectional study. Parents of a total of 3,676 fourth, seventh, and tenth graders from classrooms in 12 communities in Southern California completed questionnaires that characterized the children's histories of respiratory illness and associated risk factors. The prevalences of bronchitis, chronic phlegm, and chronic cough were investigated among children with a history of asthma, wheeze without diagnosed asthma, and neither wheeze nor asthma. Average ambient annual exposure to ozone, particulate matter (PM(10) and PM(2.5); [less than/equal to] 10 microm and < 2.5 microm in aerodynamic diameter, respectively), acid vapor, and nitrogen dioxide (NO(2)) was estimated from monitoring stations in each community. Positive associations between air pollution and bronchitis and phlegm were observed only among children with asthma. As PM(10) increased across communities, there was a corresponding increase in the risk per interquartile range of bronchitis [odds ratio (OR) 1.4/19 microg/m(3); 95% confidence interval (CI), 1.1-1.8). Increased prevalence of phlegm was significantly associated with increasing exposure to all ambient pollutants except ozone. The strongest association was for NO(2), based on relative risk per interquartile range in the 12 communities (OR 2.7/24 ppb; CI, 1.4-5.3). The results suggest that children with a prior diagnosis of asthma are more likely to develop persistent lower respiratory tract symptoms when exposed to air pollution in Southern California. PMID- 10464078 TI - Examining associations between childhood asthma and traffic flow using a geographic information system. AB - Using geographic information systems (GIS) and routinely collected data, we explored whether childhood residence near busy roads was associated with asthma in a low-income population in San Diego County, California. We examined the locations of residences of 5,996 children [less than/equal to] 14 years of age who were diagnosed with asthma in 1993 and compared them to a random control series of nonrespiratory diagnoses (n = 2,284). Locations of the children's residences were linked to traffic count data at streets within 550 ft. We also examined the number of medical care visits in 1993 for children with asthma to determine if the number of visits was related to traffic flow. Analysis of the distribution of cases and controls by quintiles and by the 90th, 95th, and 99th percentiles of traffic flow at the highest traffic street, nearest street, and total of all streets within a 550-ft buffer region did not show any significantly elevated odds ratios. However, among cases, those residing near high traffic flows (measured at the nearest street) were more likely than those residing near lower traffic flows to have two or more medical care visits for asthma than to have only one visit for asthma during the year. The results of this exploratory study suggest that higher traffic flows may be related to an increase in repeated medical visits for asthmatic children. Repeated exposure to particulate matter and other air pollutants from traffic exhaust may aggravate asthmatic symptoms in individuals already diagnosed with asthma. PMID- 10464079 TI - Methylene chloride poisoning in a cabinet worker. AB - More than a million workers are at risk for methylene chloride exposure. Aerosol sprays and paint stripping may also cause significant nonoccupational exposures. After methylene chloride inhalation, significant amounts of carbon monoxide are formed in vivo as a metabolic by-product. Poisoning predominantly affects the central nervous system and results from both carboxyhemoglobin formation and direct solvent-related narcosis. In this report, we describe a case of methylene chloride intoxication probably complicated by exogenous carbon monoxide exposure. The worker's presentation of intermittent headaches was consistent with both methylene chloride intoxication and carbon monoxide poisoning. The exposures and symptoms were corroborated by elevated carboxyhemoglobin saturations and a workplace inspection that documented significant exposures to both methylene chloride and carbon monoxide. When both carbon monoxide and methylene chloride are inhaled, additional carboxyhemoglobin formation is expected. Preventive efforts should include education, air monitoring, and periodic carboxyhemoglobin determinations. Methylene chloride should never be used in enclosed or poorly ventilated areas because of the well-documented dangers of loss of consciousness and death. PMID- 10464080 TI - Nickel carcinogenesis, mutation, epigenetics, or selection. PMID- 10464082 TI - Response from lieberman and colleagues AB - Respond on comments on Lieberman's article: Cyclosiloxanes Produce Fatal Liver and Lung Damage in Mice. Environ Health Perspect 107:161-165 PMID- 10464081 TI - Letter re: "Cyclosiloxanes produce fatal liver and lung damage in mice". AB - Comments on Lieberman's article: Cyclosiloxanes Produce Fatal Liver and Lung Damage in Mice. Environ Health Perspect 107:161-165 PMID- 10464083 TI - Nanotechnology: thinking small. AB - Nanotechnology--building devices on the atomic scale--may unleash some big scientific advances early in the new millennium. Last January in Arlington, Virginia, nearly 100 representatives from academia, industry, and government laid out the general goals for the next decade of nanotechnology research by U.S. government agencies. Some predict that the potentially rich opportunities in this field may trigger a nanotechnology initiative in the federal budget request for Fiscal Year 2001. In the meantime, Congress has asked the NIEHS to explore how nanotechnology might be used to address environmental heath problems. PMID- 10464084 TI - Where the chips fall: environmental health in the semiconductor industry. AB - Three recent lawsuits are focusing public attention on the environmental and occupational health effects of the world's largest and fastest growing manufacturing sector-the $150 billion semiconductor industry. The suits allege that exposure to toxic chemicals in semiconductor manufacturing plants led to adverse health effects such as miscarriage and cancer among workers. To manufacture computer components, the semiconductor industry uses large amounts of hazardous chemicals including hydrochloric acid, toxic metals and gases, and volatile solvents. Little is known about the long-term health consequences of exposure to chemicals by semiconductor workers. According to industry critics, the semiconductor industry also adversely impacts the environment, causing groundwater and air pollution and generating toxic waste as a by-product of the semiconductor manufacturing process. In contrast, the U.S. Bureau of Statistics shows the semiconductor industry as having a worker illness rate of about one third of the average of all manufacturers, and advocates defend the industry, pointing to recent research collaborations and product replacement as proof that semiconductor manufacturers adequately protect both their employees and the environment. PMID- 10464085 TI - Answering the endocrine test questions. AB - Evidence suggesting that certain chemicals may bind to endogenous hormone receptors and disturb normal endocrine functioning, thereby increasing the risk of reproductive problems and cancer in humans, has led to international efforts to screen chemicals for endocrine activity and potential health effects. The U.S. Environmental Protection Agency (EPA) has recommended that some 87,000 commercial chemicals for which there currently are inadequate toxicity data be evaluated. In December 1998, the EPA's Endocrine Disruptor Screening and Testing Advisory Committee (EDSTAC) recommended the use of a tiered system of screening and testing assays to sequentially eliminate chemicals for which further testing is deemed unnecessary. The first step toward implementing such a system-validation of the tests to be used-is presenting some challenges, however, with stakeholders disagreeing over which tests to validate, how extensively to validate them, and how much it will cost. PMID- 10464087 TI - Progress and achievement program PMID- 10464086 TI - TCE treatment pasta-bilities. AB - Monsanto's "Lasagna" process uses layers of treatment zones spaced between buried electrodes to remove trichloroethylene (TCE) from contaminated soil and groundwater. TCE is used primarily as a metal degreaser as well as in products such as dyes, printing ink, and paint. TCE can eventually make its way into the environment and is prevalent in the water and soil of industrialized nations. Although TCE breaks down in a few days when released into the atmosphere, it degrades much more slowly in soil, taking months or years. Moreover, it is often broken down by microbes into toxic substances such as vinylidene chloride (a suspected human carcinogen) and vinyl chloride (a known human carcinogen). The Lasagna process is based on the principle of electro-osmosis, in which an electric current draws water from low--permeability soils such as clays, silts, and fine sands. To remove TCE from contaminated soils, Monsanto scientists added layers of filtering media, which attack the contaminant as it is pulled from electrode to electrode. The technology has been tested at the Paducah Gaseous Diffusion Plant in western Kentucky, where it removed over 98% of TCE from contaminated soil. PMID- 10464088 TI - Global optimization of clusters, crystals, and biomolecules. AB - Finding the optimal solution to a complex optimization problem is of great importance in many fields, ranging from protein structure prediction to the design of microprocessor circuitry. Some recent progress in finding the global minima of potential energy functions is described, focusing on applications of the simple "basin-hopping" approach to atomic and molecular clusters and more complicated hypersurface deformation techniques for crystals and biomolecules. These methods have produced promising results and should enable larger and more complex systems to be treated in the future. PMID- 10464089 TI - Temperature Dependence of the Half-Integer Magnetic Flux Quantum. AB - The temperature dependence of the half-integer magnetic flux quantum effect in thin-film tricrystal samples of the high-critical-temperature cuprate superconductor YBa(2)Cu(3)O(7-delta) was measured and found to persist from a temperature of 0.5 kelvin through a critical temperature of about 90 kelvin, with no change in total flux. This result implies that d-wave symmetry pairing predominates in this cuprate, with a small component of time-reversal symmetry breaking, if any, over the entire temperature range. PMID- 10464090 TI - Inhibition of Crystallite Growth in the Sol-Gel Synthesis of Nanocrystalline Metal Oxides. AB - Crystal growth upon firing of hydrous transition metal oxide gels can be effectively inhibited by replacing the surface hydroxyl group before firing with another functional group that does not condense and that can produce small, secondary-phase particles that restrict advancing of grain boundaries at elevated temperatures. Accordingly, fully crystallized SnO(2), TiO(2), and ZrO(2) materials with mean crystallite sizes of approximately 20, 50, and 15 angstroms, respectively, were synthesized by replacing the hydroxyl group with methyl siloxyl before firing at 500 degrees C. An ultrasensitive SnO(2)-based chemical sensor resulting from the microstructural miniaturization was demonstrated. PMID- 10464091 TI - Asteroidal water within fluid inclusion-bearing halite in an H5 chondrite, Monahans (1998) AB - Crystals of halite and sylvite within the Monahans (1998) H5 chondrite contain aqueous fluid inclusions. The fluids are dominantly sodium chloride-potassium chloride brines, but they also contain divalent cations such as iron, magnesium, or calcium. Two possible origins for the brines are indigenous fluids flowing within the asteroid and exogenous fluids delivered into the asteroid surface from a salt-containing icy object. PMID- 10464092 TI - Origin of graphitic carbon and pentlandite in matrix olivines in the Allende meteorite. AB - Matrix olivines in the Allende carbonaceous chondrite are believed to have formed by condensation processes in the primitive solar nebula. However, transmission electron microscope observations of numerous matrix olivines show that they contain abundant, previously unrecognized, nanometer-sized inclusions of pentlandite and poorly graphitized carbon. Neither of these phases would have been stable at the high-temperature conditions required to condense iron-rich olivine in the solar nebula. The presence of these inclusions is consistent with formation of the olivines by parent body processes that involved overgrowth of fine-grained organic materials and sulfides in the precursor matrix materials. PMID- 10464093 TI - Equatorius: a new hominoid genus from the Middle Miocene of Kenya. AB - A partial hominoid skeleton just older than 15 million years from sediments in the Tugen Hills of north central Kenya mandates a revision of the hominoid genus Kenyapithecus, a possible early member of the great ape-human clade. The Tugen Hills specimen represents a new genus, which also incorporates all material previously referable to Kenyapithecus africanus. The new taxon is derived with respect to earlier Miocene hominoids but is primitive with respect to the younger species Kenyapithecus wickeri and therefore is a late member of the stem hominoid radiation in the East African Miocene. PMID- 10464094 TI - Fossil Plants and Global Warming at the Triassic-Jurassic Boundary. AB - The Triassic-Jurassic boundary marks a major faunal mass extinction, but records of accompanying environmental changes are limited. Paleobotanical evidence indicates a fourfold increase in atmospheric carbon dioxide concentration and suggests an associated 3 degrees to 4 degrees C "greenhouse" warming across the boundary. These environmental conditions are calculated to have raised leaf temperatures above a highly conserved lethal limit, perhaps contributing to the >95 percent species-level turnover of Triassic-Jurassic megaflora. PMID- 10464095 TI - Gene expression profile of aging and its retardation by caloric restriction. AB - The gene expression profile of the aging process was analyzed in skeletal muscle of mice. Use of high-density oligonucleotide arrays representing 6347 genes revealed that aging resulted in a differential gene expression pattern indicative of a marked stress response and lower expression of metabolic and biosynthetic genes. Most alterations were either completely or partially prevented by caloric restriction, the only intervention known to retard aging in mammals. Transcriptional patterns of calorie-restricted animals suggest that caloric restriction retards the aging process by causing a metabolic shift toward increased protein turnover and decreased macromolecular damage. PMID- 10464096 TI - Dual function of the selenoprotein PHGPx during sperm maturation. AB - The selenoprotein phospholipid hydroperoxide glutathione peroxidase (PHGPx) changes its physical characteristics and biological functions during sperm maturation. PHGPx exists as a soluble peroxidase in spermatids but persists in mature spermatozoa as an enzymatically inactive, oxidatively cross-linked, insoluble protein. In the midpiece of mature spermatozoa, PHGPx protein represents at least 50 percent of the capsule material that embeds the helix of mitochondria. The role of PHGPx as a structural protein may explain the mechanical instability of the mitochondrial midpiece that is observed in selenium deficiency. PMID- 10464097 TI - Eutrophication, Fisheries, and Consumer-Resource Dynamics in Marine Pelagic Ecosystems. AB - Anthropogenic nutrient enrichment and fishing influence marine ecosystems worldwide by altering resource availability and food-web structure. Meta-analyses of 47 marine mesocosm experiments manipulating nutrients and consumers, and of time series data of nutrients, plankton, and fishes from 20 natural marine systems, revealed that nutrients generally enhance phytoplankton biomass and carnivores depress herbivore biomass. However, resource and consumer effects attenuate through marine pelagic food webs, resulting in a weak coupling between phytoplankton and herbivores. Despite substantial physical and biological variability in marine pelagic ecosystems, alterations of resource availability and consumers result in general patterns of community change. PMID- 10464098 TI - Selection-Based Biodiversity at a Small Spatial Scale in a Low-Dispersing Insular Bird. AB - The blue tit is a highly mobile small passerine found in deciduous and evergreen oaks. In mainland populations, gene flow results in local maladaptive timing of breeding in evergreen oak forests, the rarer habitat. However, on the island of Corsica, two populations only 25 kilometers apart are highly specialized and differ between the two habitat types in breeding and morphological traits. In contrast to theoretical predictions about the homogenizing effects of gene flow, this highlights evolutionary consequences of habitat diversification and isolation at a small spatial scale in insular organisms, which should be taken into account in conservation policies. PMID- 10464099 TI - GTP binding by class II transactivator: role in nuclear import. AB - Class II transactivator (CIITA) is a global transcriptional coactivator of human leukocyte antigen-D (HLA-D) genes. CIITA contains motifs similar to guanosine triphosphate (GTP)-binding proteins. This report shows that CIITA binds GTP, and mutations in these motifs decrease its GTP-binding and transactivation activity. Substitution of these motifs with analogous sequences from Ras restores CIITA function. CIITA exhibits little GTPase activity, yet mutations in CIITA that confer GTPase activity reduce transcriptional activity. GTP binding by CIITA correlates with nuclear import. Thus, unlike other GTP-binding proteins, CIITA is involved in transcriptional activation that uses GTP binding to facilitate its own nuclear import. PMID- 10464100 TI - Rapid adaptation in visual cortex to the structure of images. AB - Complex cells in striate cortex of macaque showed a rapid pattern-specific adaptation. Adaptation made cells more sensitive to orientation change near the adapting orientation. It reduced correlations among the responses of populations of cells, thereby increasing the information transmitted by each action potential. These changes were brought about by brief exposures to stationary patterns, on the time scale of a single fixation. Thus, if successive fixations expose neurons' receptive fields to images with similar but not identical structure, adaptation will remove correlations and improve discriminability. PMID- 10464101 TI - Altered cochlear fibrocytes in a mouse model of DFN3 nonsyndromic deafness. AB - DFN3, an X chromosome-linked nonsyndromic mixed deafness, is caused by mutations in the BRN-4 gene, which encodes a POU transcription factor. Brn-4-deficient mice were created and found to exhibit profound deafness. No gross morphological changes were observed in the conductive ossicles or cochlea, although there was a dramatic reduction in endocochlear potential. Electron microscopy revealed severe ultrastructural alterations in cochlear spiral ligament fibrocytes. The findings suggest that these fibrocytes, which are mesenchymal in origin and for which a role in potassium ion homeostasis has been postulated, may play a critical role in auditory function. PMID- 10464102 TI - Long-term depression in hippocampal interneurons: joint requirement for pre- and postsynaptic events. AB - Long-term depression (LTD) is a well-known form of synaptic plasticity of principal neurons in the mammalian brain. Whether such changes occur in interneurons is still controversial. CA3 hippocampal interneurons expressing Ca2+ permeable AMPA receptors exhibited LTD after tetanic stimulation of CA3 excitatory inputs. LTD was independent of NMDA receptors and required both Ca2+ influx through postsynaptic AMPA receptors and activation of presynaptic mGluR7 like receptors. These results point to the capability of interneurons to undergo plastic changes of synaptic strength through joint activation of pre- and postsynaptic glutamate receptors. PMID- 10464127 TI - New antiviral agents for viral hepatitis: our cup runneth over but is only half full. PMID- 10464128 TI - Lack of effect of intravenous administration on time to respond to azathioprine for steroid-treated Crohn's disease. North American Azathioprine Study Group. AB - BACKGROUND & AIMS: Azathioprine is effective for Crohn's disease but acts slowly. A loading dose may decrease the time to response. METHODS: A placebo-controlled study was conducted in patients with active Crohn's disease despite prednisone treatment. Patients were randomized to a 36-hour infusion of azathioprine, 40 mg/kg (51 patients), or placebo (45 patients) followed by oral azathioprine, 2 mg/kg, for 16 weeks. Prednisone was tapered over 5 weeks. The primary outcome measure was complete remission at week 8, defined by discontinuation of prednisone and a Crohn's Disease Activity Index of 49 years (RR, 1.4; 95% CI, 1.1-1.8; P = 0.01). CONCLUSIONS: The results of transplantation for HCV-related disease are compromised by a significant risk of cirrhosis, although 5-year survival is satisfactory. Genotype 1b, age, and absence of pretransplantation coinfection by HBV are risk factors for recurrent HCV. PMID- 10464138 TI - Prognostic value of early measurements of portal pressure in acute variceal bleeding. AB - BACKGROUND & AIMS: Variceal bleeding is the most important complication of portal hypertension. However, the relationship between the increase in portal pressure and the outcome of variceal bleeding has not been well defined. METHODS: We measured the hepatic venous pressure gradient (HVPG) of 65 cirrhotic patients with acute variceal hemorrhage, early after admission (20.6 +/- 15.6 hours). RESULTS: Twenty-three patients had a poor evolution (failure to control bleeding or early variceal rebleeding), and 42 had an uneventful evolution. The only variable associated with outcome was the HVPG, which was higher in patients with a poor evolution (23.7 +/- 6.1 vs. 19.2 +/- 3.3 mm Hg; P < 0.0004). This was confirmed by multivariate analysis. HVPG was >/=20 mm Hg in 19 of 23 patients with poor evolution vs. 12 of 42 patients with uneventful evolution (P < 0.0001). An initial HVPG of >/=20 mm Hg was associated with a significantly longer intensive care unit stay (7 +/- 5 vs. 4 +/- 2 days; P < 0.02), longer hospital stay (19 +/- 10 vs. 14 +/- 6 days; P < 0.02), greater transfusion requirements (9.0 +/- 7.7 vs. 4.7 +/- 3.2 UU; P < 0.007), and a worse actuarial probability of survival (1-year mortality, 64% vs. 20%; P < 0.002). CONCLUSIONS: Early measurement of HVPG in cirrhotic patients during acute variceal bleeding provides useful prognostic information on the evolution of the bleeding episode and long term survival. PMID- 10464139 TI - Prevalence and ethnic differences in gallbladder disease in the United States. AB - BACKGROUND & AIMS: Gallbladder disease is one of the most common conditions in the United States, but its true prevalence is unknown. A national population based survey was performed to determine the age, sex, and ethnic distribution of gallbladder disease in the United States. METHODS: The third National Health and Nutrition Examination Survey (NHANES III) conducted gallbladder ultrasonography among a representative U.S. sample of more than 14, 000 persons. The diagnosis of gallbladder disease by detection of gallstones or cholecystectomy was made with excellent reproducibility. RESULTS: An estimated 6.3 million men and 14.2 million women aged 20-74 years had gallbladder disease. Age-standardized prevalence was similar for non-Hispanic white (8. 6%) and Mexican American (8.9%) men, and both were higher than non-Hispanic black men (5.3%). These relationships persisted with multivariate adjustment. Among women, age-adjusted prevalence was highest for Mexican Americans (26.7%) followed by non-Hispanic whites (16.6%) and non Hispanic blacks (13.9%). Among women, multivariate adjustment reduced the risk of gallbladder disease for both Mexican Americans and non-Hispanic blacks compared with non-Hispanic whites. CONCLUSIONS: More than 20 million persons have gallbladder disease in the United States. Ethnic differences in gallbladder disease prevalence differed according to sex and were only partly explained by known risk factors. PMID- 10464140 TI - Manganese deposition in basal ganglia structures results from both portal systemic shunting and liver dysfunction. AB - BACKGROUND & AIMS: Manganese (Mn) deposition could be responsible for the T(1) weighted magnetic resonance signal hyperintensities observed in cirrhotic patients. These experiments were designed to assess the regional specificity of the Mn increases as well as their relationship to portal-systemic shunting or hepatobiliary dysfunction. METHODS: Mn concentrations were measured in (1) brain samples from basal ganglia structures (pallidum, putamen, caudate nucleus) and cerebral cortical structures (frontal, occipital cortex) obtained at autopsy from 12 cirrhotic patients who died in hepatic coma and from 12 matched controls; and from (2) brain samples (caudate/putamen, globus pallidus, frontal cortex) from groups (n = 8) of rats either with end-to-side portacaval anastomosis, with biliary cirrhosis, or with fulminant hepatic failure as well as from sham operated and normal rats. RESULTS: Mn content was significantly increased in frontal cortex (by 38%), occipital cortex (by 55%), pallidum (by 186%), putamen (by 66%), and caudate (by 54%) of cirrhotic patients compared with controls. Brain Mn content did not correlate with patient age, etiology of cirrhosis, or history of chronic hepatic encephalopathy. In cirrhotic and portacaval-shunted rats, Mn content was increased in pallidum (by 27% and 57%, respectively) and in caudate/putamen (by 57% and 67%, respectively) compared with control groups. Mn concentration in pallidum was significantly higher in portacaval-shunted rats than in cirrhotic rats. No significant changes in brain Mn concentrations were observed in rats with acute liver failure. CONCLUSIONS: These findings suggest that brain Mn deposition results both from portal-systemic shunting and from liver dysfunction. PMID- 10464141 TI - Cytokine polymorphisms associated with clinical features and treatment outcome in type 1 autoimmune hepatitis. AB - BACKGROUND & AIMS: Polymorphisms that control cytokine production can affect immunoregulation. The frequency and consequences of these polymorphisms in type 1 autoimmune hepatitis were determined. METHODS: DNA samples from 155 patients and 102 ethnically similar normal individuals were assessed by polymerase chain reaction for polymorphisms of 4 different cytokine-producing genes. RESULTS: Only genotypes associated with the guanine to adenine substitution at position -308 of the tumor necrosis factor gene occurred more commonly in patients than in normal subjects (56% vs. 26%; P < 0.001). Patients with this polymorphism had the HLA DRB1*0301 allele (81% vs. 10%; P < 0.000001) and A1-B8-DRB1*0301 (66% vs. 0%; P < 0.000001) phenotype more frequently and HLA DRB1*04 alleles less often (24% vs. 67%; P < 0.000001). They also entered remission less commonly (56% vs. 78%; P = 0.01), had treatment failure more often (20% vs. 7%; P = 0.03), and developed cirrhosis more frequently (40% vs. 19%; P = 0.05). These latter differences, however, were not statistically significant by adjusted P value. CONCLUSIONS: A polymorphism of the tumor necrosis factor gene occurs more commonly in patients with type 1 autoimmune hepatitis than in normal subjects; it is associated with a poorer response to corticosteroids. The polymorphism may be inherited as part of the extended A1-B8-DRB1*0301 haplotype and may affect both disease expression and behavior. PMID- 10464142 TI - Exon-intron organization of the human multidrug-resistance protein 2 (MRP2) gene mutated in Dubin-Johnson syndrome. AB - BACKGROUND & AIMS: The Dubin-Johnson syndrome is characterized by conjugated hyperbilirubinemia and by impaired secretion of anionic conjugates from hepatocytes into bile. Absence of the multidrug-resistance protein 2 (MRP2; symbol ABCC2), an adenosine triphosphate-dependent conjugate export pump, from the hepatocyte canalicular membrane is the molecular basis of this syndrome. The aim of this study was the elucidation of all exon-intron boundaries of the MRP2 gene as a prerequisite for the analysis of mutations in patients with Dubin Johnson syndrome. METHODS: Exon-intron boundaries of MRP2 were determined, and the amplified exons were screened for mutations. Immunofluorescence microscopy served to localize the MRP2 protein in human liver. RESULTS: The human MRP2 gene is approximately 45 kilobases long; it contains 32 exons and a high proportion of class 0 introns. In 2 patients with Dubin-Johnson syndrome, we detected a nonsense mutation at codon 1066 and a 6-nucleotide deletion mutation affecting codons 1392-1394. The MRP2 protein was absent from the canalicular membrane of both patients. CONCLUSIONS: The mutations detected so far show that various mutations in the MRP2 gene can lead to the Dubin-Johnson syndrome. The exon intron boundaries established in this article will facilitate the analysis of additional mutations in the MRP2 gene. PMID- 10464143 TI - Interleukin 1beta protects mice from Fas-mediated hepatocyte apoptosis and death. AB - BACKGROUND & AIMS: Fas-mediated apoptosis is one of the major death processes of hepatocytes in liver diseases. The aim of this study was to determine whether interleukin (IL)-1beta regulates the Fas-mediated apoptotic process of differentiated hepatocytes in vivo. METHODS: IL-1beta was injected into Balb/cA mice 5 hours before lethal challenge with agonistic anti-Fas administration. Survival and hepatocyte apoptotic process of these mice were examined. RESULTS: IL-1beta pretreatment prolonged animal survival in a dose-dependent manner, and 500 ng of IL-1beta completely protected mice from lethality. Both serum alanine aminotransferase value and hepatic DNA fragmentation were significantly suppressed by IL-1beta pretreatment. IL-1beta affected neither hepatic distribution of anti-Fas antibody nor Fas expression levels on hepatocytes but significantly suppressed Fas-induced activation of hepatic caspase 3-like protease. Suppression of Fas-induced activation of the caspase by IL-1beta was diminished by coadministration with D-galactosamine and reversed by coinjection with an excess amount of uridine. CONCLUSIONS: These results suggest that IL 1beta suppresses Fas-mediated hepatocyte apoptosis by inducing molecule(s) that suppress the apoptosis control machinery upstream of caspase 3. This observation raises the possibility that IL-1beta acts as a negative regulator of Fas-mediated hepatocyte apoptosis during liver injury. PMID- 10464144 TI - Hepatocyte apoptosis after bile duct ligation in the mouse involves Fas. AB - BACKGROUND & AIMS: Cholestatic liver injury results from the intrahepatic accumulation of toxic bile salts. Toxic bile salt-induced hepatocyte apoptosis in vitro is Fas dependent. The aim of this study was to ascertain if hepatocyte apoptosis in vivo during cholestasis is Fas dependent. METHODS: Studies were performed in bile duct-ligated (BDL) Fas-deficient lpr (lymphoproliferation) and wild-type mice. RESULTS: Hepatocyte apoptosis was the predominant mechanism of cell death as determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling and trypan blue assays to quantitate apoptosis and necrosis. The mechanisms of hepatocyte apoptosis were dependent on the presence or absence of the Fas receptor and the duration of BDL. After BDL of 3 days' duration, increased hepatocyte apoptosis occurred only in wild-type but not lpr mice, indicating the apoptosis was Fas dependent. In contrast, after BDL of >/=7 days, hepatocyte apoptosis also occurred in lpr animals consistent with a Fas-independent mechanism of apoptosis. Hepatocyte apoptosis in BDL lpr mice was associated with an increase in Bax expression and Bax association with mitochondria. CONCLUSIONS: During extrahepatic cholestasis, hepatocyte apoptosis is mediated by Fas. However, in the absence of the Fas receptor, additional mechanisms of hepatocyte apoptosis occur. Inhibition of multiple apoptotic pathways is necessary to attenuate chronic cholestatic liver injury. PMID- 10464145 TI - 3-Hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) induce hepatic expression of the phospholipid translocase mdr2 in rats. AB - BACKGROUND & AIMS: Biliary cholesterol secretion is coupled to that of phospholipids in a process controlled by mdr2 P-glycoprotein activity and bile salt secretion. Statins, the 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, have been shown to affect hepatobiliary lipid secretion in rats. The aim of this study was to relate the effects of statins on bile formation to the expression of mdr2 and other hepatic adenosine triphosphate dependent transport proteins involved in bile formation in rats. METHODS: Rats received simvastatin- or pravastatin-containing chow continuously for 5 days. In one group of rats, simvastatin treatment was withdrawn 9-12 hours before the end of the experiment to induce biliary cholesterol hypersecretion (rebound). Bile and liver tissue were collected for lipid analysis, and hepatic messenger RNA (mRNA) and protein levels were studied by reverse-transcription polymerase chain reaction, immunoblotting, and immunohistochemistry. RESULTS: Simvastatin feeding did not alter biliary bile salt secretion. Secretion of phospholipids and cholesterol was stimulated by 74% and 90%, respectively, in the simvastatin continuous group and by 72% and 235%, respectively, in the rebound group compared with controls. mdr2 mRNA levels increased only in the continuous group. mdr2 protein levels increased in both simvastatin-fed groups. Induction was most pronounced in periportal hepatocytes. mdr1b mRNA levels were moderately increased in both simvastatin-fed groups. Levels of other hepatic transport proteins did not change. Similar results were obtained in pravastatin-fed rats. CONCLUSIONS: Statins increase expression of mdr2 and mdr1b in rats, revealing a novel effect of these commonly used drugs. PMID- 10464146 TI - Localization and function of the organic anion-transporting polypeptide Oatp2 in rat liver. AB - BACKGROUND & AIMS: Multispecific organic anion-transporting polypeptides (Oatps) are involved in the transcellular movement of amphipathic compounds in many tissues including the liver, kidney, and blood-brain barrier. Recently, a high affinity digoxin transporter (Oatp2) was cloned from rat brain and shown to be also expressed in the liver. METHODS: We investigated the cellular and subcellular distribution of Oatp2 in rat liver by in situ hybridization technology and immunofluorescence microscopy and compared its substrate specificity with that of Oatp1 in complementary RNA-injected Xenopus laevis oocytes. RESULTS: The results show a selective basolateral (sinusoidal) expression of Oatp2 in midzonal to perivenous hepatocytes, but not in periportal or the innermost layer of perivenous hepatocytes. Common substrates of both Oatp1 and Oatp2 include bile salts, steroid conjugates, thyroid hormones (T3, T4), ouabain, and the endothelin receptor antagonist BQ-123 (Michaelis constants: Oatp1, approximately 600 micromol/L; Oatp2, approximately 30 micromol/L). Other organic anions including sulfolithotaurocholate, bilirubin monoglucuronide, and sulfobromophthalein were transported only by Oatp1. CONCLUSIONS: These results provide definite evidence for the partially overlapping and partially selective substrate specificities of Oatp1 and Oatp2. The unique acinar distribution of Oatp2 might indicate that it represents a high-affinity "backup" system for complete hepatocellular removal of certain cholephilic substances from portal blood plasma. PMID- 10464147 TI - Biliary fibrosis associated with altered bile composition in a mouse model of erythropoietic protoporphyria. AB - BACKGROUND & AIMS: Reduced activity of ferrochelatase in erythropoietic protoporphyria (EPP) results in protoporphyrin (PP) accumulation in erythrocytes and liver. Liver disease may occur in patients with EPP, some of whom develop progressive liver failure that necessitates transplantation. We investigated the mechanisms underlying EPP-associated liver disease in a mouse model of EPP. METHODS: Liver histology, indicators of lipid peroxidation, plasma parameters of liver function, and bile composition were studied in mice homozygous (fch/fch) for a point mutation in the ferrochelatase gene and in heterozygous (fch/+) and wild-type (+/+) mice. RESULTS: Microscopic examination showed bile duct proliferation and biliary fibrosis with portoportal bridging in fch/fch mice. PP content was 130-fold increased, and thiobarbituric acid-reactive substances (+30%) and conjugated dienes (+75%) were slightly higher in fch/fch than in fch/+ and +/+ livers. Levels of hepatic thiols (-12%) and iron (-52%) were reduced in fch/fch livers. Liver enzymes and plasma bilirubin were markedly increased in the homozygotes. Plasma bile salt levels were 80 times higher in fch/fch than in fch/+ and +/+ mice, probably related to the absence of the Na(+)-taurocholate cotransporting protein (Ntcp) in fch/fch liver. Paradoxically, bile flow was not impaired and biliary bile salt secretion was 4 times higher in fch/fch mice than in controls. Up-regulation of the intestinal Na(+)-dependent bile salt transport system in fch/fch mice may enhance efficiency of bile salt reabsorption. The bile salt/lipid ratio and PP content of fch/fch bile were increased 2-fold and 85 fold, respectively, compared with +/+, whereas biliary glutathione was reduced by 90%. Similar effects on bile formation were caused by griseofulvin-induced inhibition of ferrochelatase activity in control mice. CONCLUSIONS: Bile formation is strongly affected in mice with impaired ferrochelatase activity. Rather than peroxidative processes, formation of cytotoxic bile with high concentrations of bile salts and PP may cause biliary fibrosis in fch/fch mice by damaging bile duct epithelium. PMID- 10464148 TI - Ethanol diet increases the sensitivity of rats to pancreatitis induced by cholecystokinin octapeptide. AB - BACKGROUND & AIMS: Although alcoholism is a major cause of pancreatitis, the pathogenesis of this disorder remains obscure. Failure to produce experimental alcoholic pancreatitis suggests that ethanol may only increase predisposition to pancreatitis. This study sought to develop a model of ethanol pancreatitis by determining if an ethanol diet sensitizes rats to pancreatitis caused by cholecystokinin octapeptide (CCK-8). METHODS: Rats were fed intragastrically either control or ethanol diet for 2 or 6 weeks. The animals were then infused for 6 hours with either saline or CCK-8 at a dose of 3000 pmol. kg(-1). h(-1), which by itself did not induce pancreatitis. The following parameters were measured: serum amylase and lipase levels, pancreatic weight, inflammatory infiltration, number of apoptotic acinar cells, pancreatic messenger RNA (mRNA) expression of cytokines and chemokines, and nuclear factor (NF)-kappaB activity. RESULTS: All measures of pancreatitis, as well as NF-kappaB activity and mRNA expression for tumor necrosis factor alpha, interleukin 6, monocyte chemotactic protein 1, macrophage inflammatory protein 2, and inducible nitric oxide synthase, were significantly increased only in rats treated with ethanol plus CCK 8. CONCLUSIONS: An ethanol diet sensitizes rats to pancreatitis caused by CCK-8. The combined action of ethanol and CCK-8 results in NF-kappaB activation and up regulation of proinflammatory cytokines and chemokines in the pancreas. These mechanisms may contribute to the development of alcoholic pancreatitis. PMID- 10464149 TI - Interstitial trypsinogen release and its relevance to the transformation of mild into necrotizing pancreatitis in rats. AB - BACKGROUND & AIMS: Intracellular activation of trypsinogen is currently believed to initiate pancreatitis. Factors responsible for the progression of mild to necrotizing pancreatitis are poorly understood. This study evaluated the significance of interstitial protease release and activation in this process. METHODS: In rats with cerulein-induced pancreatitis, concentrations of trypsinogen and its activation peptide TAP were measured in lymph and blood, and pancreatic injury was determined. Activation of extracellular trypsinogen was induced by intravenous infusion of enterokinase, which does not enter the acinar cell. Gabexate mesilate (acinar cell permeable) or soybean trypsin inhibitor (acinar cell nonpermeable) was administered to distinguish the effects of intracellular or extracellular protease activation. RESULTS: In cerulein pancreatitis, trypsinogen levels increased prominently and were highest in lymph and portal vein blood, whereas TAP increments were modest. Combined cerulein/enterokinase infusions resulted in marked TAP increases in lymph and blood and in severe necrohemorrhagic pancreatitis. Gabexate mesilate as well as soybean trypsin inhibitor significantly decreased TAP levels in both lymph and blood and reduced pancreatic injury, with no significant differences between groups. CONCLUSIONS: In secretagogue-induced pancreatitis, large amounts of trypsinogen are present in the interstitium and drain via the portal and lymphatic circulation. Activation of this extracellular trypsinogen induces hemorrhagic necrosis in a setting of mild edematous pancreatitis. This phenomenon may be the central event in the progression to fulminant necrotizing pancreatitis. PMID- 10464150 TI - What's so (Co)stimulating about the intestinal epithelium? PMID- 10464151 TI - Searching for the needle in the haystack: another candidate needle in autoimmune hepatitis? PMID- 10464152 TI - Bile acids and hepatocyte apoptosis: living/leaving life in the Fas lane. PMID- 10464153 TI - The problem of delayed emesis after cancer chemotherapy: a possible new treatment. PMID- 10464154 TI - Does interferon therapy prevent hepatocellular carcinoma in patients with chronic hepatitis C? PMID- 10464155 TI - Angiotensin II receptor antagonists in the pharmacological therapy of portal hypertension: a caution. PMID- 10464156 TI - Acid Related Disease: Biology and Treatment. PMID- 10464157 TI - Antibodies against human heat-shock protein (hsp) 60 and mycobacterial hsp65 differ in their antigen specificity and complement-activating ability. AB - Although complement activation appears to have an important role both in the early and late phases of atherosclerosis, the exact mechanism of the initiation of this activation is still unknown. Since injuries of the endothelial cells are known to result in increased stress-protein expression we tested the complement activating ability of recombinant human 60 kDa heat-shock protein (hsp60). Human hsp60 was found to activate the complement system in normal human serum in a dose dependent manner. Activation took place through the classical pathway. The lack of complement activation in agammaglobulinemic serum indicates that the classical pathway is triggered by anti-hsp60 antibodies. Hsp60 activated complement in the sera of 74 patients with coronary heart disease as well, and a strong positive correlation (r = 0.459, P < 0.0001) was found between the extent of complement activation and the level of anti-hsp60 IgG antibodies but there was no correlation to the level of anti-hsp65 IgG antibodies. Further distinction between anti-hsp60 and anti-hsp65 antibodies was obtained from competitive ELISA experiments: binding of anti-hsp60 antibodies to hsp60-coated plates was inhibited only by recombinant hsp60 and vice versa. Our present findings indicate that anti-hsp60 and anti-hsp65 antibodies are distinct, showing only partial cross-reactivity. Since complement activation plays an important role in the development of atherosclerosis and the levels of complement-activating anti-hsp60 antibodies are elevated in atherosclerosis-related diseases, our present findings may have important pathological implications. PMID- 10464158 TI - Protein tyrosine kinase Lyn mediates apoptosis induced by topoisomerase II inhibitors in DT40 cells. AB - Several sets of non-receptor protein tyrosine kinases (PTK) play important roles in apoptosis induced by various extracellular stresses. Anti-cancer drugs induce cellular DNA damage and cytotoxic events, leading to apoptotic cell death. We utilized the established chicken B cell line, DT40 cells and their derived mutants, lacking the respective PTK [DT40/Syk(-), DT40/Lyn(-) and DT40/Btk(-)], to examine a role of these PTK in apoptotic processes induced by anti-cancer drugs. All anti-cancer drugs examined induced apoptosis of wild-type DT40 cells. Interestingly,DT40/Lyn(-), but not DT40/Syk(-) and DT40/Btk(-) cells, become resistant to apoptosis induced by adriamycin and etoposide, topoisomerase II (Topo II) inhibitory agents, compared to wild-type DT40 cells, as assessed by DNA fragmentation and TUNEL analyses. Ectopic expression of Fyn, another Src family member, in DT40/Lyn(-) cells restores largely the susceptibility of the cells against Topo II inhibitor-induced apoptosis. Furthermore, it was found that Topo II inhibitors activate c-Jun N-terminal kinase (JNK) slightly in both wild-type and DT40/Lyn(-) cells to similar extents. Collectively, these results suggest that Lyn is involved in Topo II inhibitor-induced apoptotic signaling in DT40 cells independent of JNK. PMID- 10464159 TI - Immobilization of glycosylphosphatidylinositol-anchored proteins inhibits T cell growth but not function. AB - Accumulating evidence suggests that proteins tethered to the plasma membrane through glycosylphosphatidylinositol (GPI) anchors share common biological properties. In the present study we demonstrate that GPI-anchored proteins regulate T cell growth. Specifically, anti-TCR-induced proliferation was profoundly inhibited by co-immobilized mAb specific for Thy-1, CD48 and Ly6A/E. However, neither IL-2 production nor the effector function of cytotoxic T lymphocytes was impaired in these circumstances. Analysis of the IL-2 receptor (IL-2R) signaling pathway revealed that the association of IL-2R beta and gamma chains with the Janus kinases, JAK1 and JAK3, was not perturbed in the presence of mAb specific for GPI-linked proteins. However, in these conditions, IL-2 mediated recruitment of IL-2Ralpha, beta and gamma chains, resulting in the formation of the high-affinity hetero-trimeric IL-2R, was inhibited. The resulting phosphorylation of JAK1 and JAK3, indicative of their activation states, was correspondingly reduced. These results characterize a novel state of T cell physiology in which effector function is maintained, in the absence of clonal expansion. A physiological role for GPI-anchored proteins in the maintenance of cellular homeostasis and function is discussed. PMID- 10464160 TI - Translocation of tyrosine-phosphorylated TCRzeta chain to glycolipid-enriched membrane domains upon T cell activation. AB - Recent studies point to glycolipid-enriched membrane (GEM) microdomains as the critical sites for TCR-mediated signal transduction. However, whether the TCR complex is localized in the GEM domain is not well-defined. In the present study, we analyzed localization of the TCR-CD3 complex in the GEM domain by isolating the GEM fraction with sucrose density gradient centrifugation. Although 10% of TCRzeta chains was localized in the GEM fraction, most of the TCR complexes were excluded from the GEM before and after T cell activation, and the amount of TCRzeta in the GEM was not increased after activation. However, the tyrosine phosphorylated form of TCRzeta was strongly concentrated in the GEM fraction upon TCR engagement. A kinetic study revealed that tyrosine phosphorylation of TCRzeta occurred initially in the Triton X-100-soluble membrane fraction followed by the accumulation of phosphorylated TCRzeta in the GEM. Thus, these results indicate that phosphorylated TCRzeta migrates into the GEM domains on T cell activation. We speculate that the GEM microdomains may function as a reservoir of activation signals from triggered TCR. PMID- 10464161 TI - Role and regulation of IL-12 in the in vivo response to staphylococcal enterotoxin B. AB - Injection of a staphylococcal superantigen (SAg) such as staphylococcal enterotoxin B (SEB) in adult mice results in cytokine production and cell proliferation which can lead to septic shock. The aim of the present work was to identify the cytokines and co-stimulatory molecules regulating the in vivo systemic release of IFN-gamma, a cytokine known to play an important role in the pathophysiology associated with bacterial infections. We demonstrate in this study that (i) in contrast to lipopolysaccharide (LPS), SEB administration induces high levels of the p70, bioactive form, of IL-12; (ii) IL-12 production in response to SEB requires both CD40-dependent signals and IFN-gamma secretion; (iii) the early systemic release of IFN-gamma (3 h post-treatment) in response to SEB is IL-12 independent, while the sustained, late response (6-9 h post treatment) requires endogenous IL-12 production; (iv) IL-12 produced during the primary SEB response (day 0) is responsible for priming cells in vivo to high IFN gamma production upon secondary challenge (day 2); (v) the priming effect of IL 12 is TCR unrelated, as SEB-primed animals secrete high levels of IFN-gamma in response to both staphylococcal enterotoxin A and LPS administered 48 h later. The ability of bacterial SAg to induce septic shock and to modulate the immune response to unrelated antigens may therefore be related to their unique capacity to induce systemic IL-12 production in vivo. These observations also help to explain why SEB-primed animals, known to express an anergic phenotype 48 h post treatment (as judged by defective IL-2 production and proliferation), nevertheless display an increased capacity to secrete the inflammatory cytokine IFN-gamma. PMID- 10464162 TI - Defective TCR signaling events in glycosylphosphatidylinositol-deficient T cells derived from paroxysmal nocturnal hemoglobinuria patients. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemolytic disorder characterized by the presence of abnormal cells of various hematopoietic cell lineages deficient in surface expression of glycosylphosphatidylinositol (GPI) anchored molecules. By analyzing T cells isolated from patients affected with PNH, it was found that ex vivo GPI-deficient CD4(+) and CD8(+) peripheral T cells display a more naive phenotype as compared to wild-type cells. In addition, in vitro proliferative responses to allogeneic antigen-presenting cells were shown to be reduced in mutant T cells. To investigate the molecular basis responsible for defective T cell activation in GPI-deficient T cells, T cell lines and T cell clones were generated from patients affected with PNH. When stimulated with anti CD3epsilon mAb, mutant cells displayed a significantly decreased activation of protein tyrosine kinase p56(lck). The decreased kinase activity was accompanied by a delayed TCR capping and internalization. Interestingly, protein tyrosine phosphorylation is not only quantitatively but also qualitatively affected, with one substrate being more intensively phosphorylated in mutant than in wild-type cells. These observations suggest that a defective activation of p56(lck) contributes to the depressed immune responses observed in GPI-deficient T cells derived from PNH patients. PMID- 10464163 TI - Prf, a novel Ets family protein that binds to the PU.1 binding motif, is specifically expressed in restricted stages of B cell development. AB - During the development of lymphocytes, expression of the Ig genes is strictly regulated in a tissue-specific manner and in a time-ordered fashion. We have previously shown that the PU.1 binding motif in the Igkappa 3' enhancer (kappaE3') and a novel Ets family protein other than PU.1 may be possibly involved in the control of V(kappa)-J(kappa) joining. In the attempt to isolate the novel Ets family protein, we have screened cDNA libraries with the yeast one hybrid method and identified a new PU.1-related factor, Prf. This novel Ets family protein is shown to interact with the PU.1 binding sequences in various promoters and enhancers, including kappaE3'. It was found that expression of the prf gene is predominant in the B-lineage cells, with the exception of immature B cells. Since Prf does not exhibit functions of transcriptional activity, this novel protein may act as an antagonist against other Ets family proteins, e.g. PU.1 and Spi-B. Possible roles of Prf with respect to the B cell differentiation are discussed. PMID- 10464164 TI - Mice lacking the transcription factor subunit Rel can clear an influenza infection and have functional anti-viral cytotoxic T cells but do not develop an optimal antibody response. AB - Rel, a haemopoietic cell-restricted member of the NF-kappaB/Rel family of transcription factors, has recently been shown to be important in the function of B and T lymphocytes. In an attempt to understand the role of this protein in the immune response, we examined the ability of Rel(-/-) mice to counter an influenza virus infection. Normal levels of virus-specific cytotoxic T cells induced in Rel(-/-) mice were able to clear virus from the lungs, albeit with somewhat delayed kinetics compared to normal mice. Rel(-/-) mice did, however, display a markedly reduced T cell proliferative response to the virus, and exhibited impaired local and systemic influenza virus-specific antibody responses. This defect was sufficient to result in an inability of vaccinated mice, but not of previously infected mice, to acquire antibody-dependent protective immunity to reinfection with the same virus. These findings establish that during the response to influenza virus, Rel function allows optimal development of humoral immunity, a role that apparently cannot be fulfilled by other NF-kappaB/Rel proteins. PMID- 10464165 TI - Distinctive roles of Fyn and Lyn in IgD- and IgM-mediated signaling. AB - Src family kinases Fyn and Lyn associate with the B cell antigen receptor (BCR). Accumulating data show that Lyn plays important roles in BCR-mediated signaling, while the role of Fyn remains obscure. Here we dissected the role of Fyn and Lyn in BCR signaling using B cells from fyn(-/-), lyn(-/-) and fyn/lyn double deficient (fyn(-/-)lyn(-/-)) mice. In contrast to previous reports, fyn(-/-) B cells were slightly hyporeactive to both anti-IgM and anti-IgD-dextran. Although lyn(-/-) B cells were hyper-reactive to anti-IgM, anti-IgD-induced proliferation was impaired in lyn(-/-) B cells. Most of the other phenotypes of fyn(-/-)lyn(-/ ) mice were similar to that of lyn(-/-) mice, except that proliferative responses of B cells to various stimuli, such as BCR cross-linking and lipopolysaccharide, were significantly lower in fyn(-/-)lyn(-/-) mice than in lyn(-/-) mice. Finally, immune responses to thymus-independent type 2 antigen were affected in these mutant mice. These observations suggest that Fyn and Lyn are involved in B cell functions, and play similar, but partly distinct, roles in BCR signaling. PMID- 10464167 TI - Virus-induced non-specific signals cause cell cycle progression of primed CD8(+) T cells but do not induce cell differentiation. AB - In this report the significance of virus-induced non-specific T cell activation was re-evaluated using transgenic mice in which about half of the CD8(+) T cells expressed a TCR specific for amino acids 33-41 of lymphocytic choriomeningitis virus glycoprotein I. This allowed tracing of cells with known specificity and priming history in an environment also containing a normal heterogeneous CD8(+) population which served as an intrinsic control. Three parameters of T cell activation were analyzed: cell cycle progression, phenotypic conversion and cytolytic activity. Following injection of the IFN inducer poly(I:C), proliferation of memory (CD44(hi)) CD8(+) T cells but no phenotypic or functional activation was observed. Following injection of an unrelated virus [vesicular stomatitis virus (VSV)], naive TCR transgenic cells did not become significantly activated with respect to any of the parameters investigated. In contrast, memory TCR transgenic cells were found to proliferate extensively early after VSV infection (day 0-3), whereas limited proliferation was observed later (day 3-6) when proliferation of non-transgenic CD8(+) T cells is maximal. This aborted response did not result from anergy to TCR stimulation, as memory TCR transgenic cells proliferated vigorously upon stimulation with their nominal peptide. Despite the massive proliferation of memory cells observed early after VSV infection, no phenotypic or functional activation was observed. Together these findings indicate that both non-specific and antigen-specific signals contribute to the initial virus-induced proliferation of CD8(+) T cells, but for further proliferation and differentiation to take place, TCR-ligand interaction is required. The implications for maintenance of T cell memory is discussed. PMID- 10464166 TI - TCR-mediated activation of allergen-specific CD45RO(+) memory T lymphocytes results in down-regulation of cell-surface CXCR4 expression and a strongly reduced capacity to migrate in response to stromal cell-derived factor-1. AB - The selective migration of functional T(h) lymphocyte subsets with different cytokine production profiles into inflamed tissue is likely to depend on the state of activation of the cells, as well as on the differential expression of various adhesion molecules and chemokine receptors. In this study, we have analyzed the effect of allergen-specific activation on the expression of the chemokine receptor CXCR4 on T lymphocytes. We show that stimulation of peripheral blood mononuclear cells from atopic patients with the allergen Der p results in down-regulation of CXCR4 surface expression on Der p-activated CD25(+)CD45RO(+) antigen-specific memory cells which was caused by a decrease in CXCR4 gene transcription and did not seem to be mediated by endogenous cytokines, such as IFN-gamma. In contrast, however, CXCR4 surface expression was enhanced on naive CD25(-)CD45RO(-) and resting CD25(-)CD45RO(+) memory T cells, as a result of the presence of endogenous IL-4, most likely produced by Der p-activated memory T cells. Antigen-specific CD25(+)CD45RO(+) T lymphocytes, purified 7 days after stimulation with Der p, had a strongly reduced capacity to migrate in response to stimulation with stromal cell-derived factor (SDF)-1, the ligand for CXCR4. Together, these results suggest that differential expression of CXCR4 on activated and resting T cells is due to the counteracting effects of TCR-mediated down-regulation and IL-4-mediated up-regulation of this chemokine receptor respectively, and furthermore indicate that antigen-activated memory T cells are unlikely to migrate into inflamed tissue in response to SDF-1. PMID- 10464168 TI - Gammadelta T cells increase with Mycobacterium avium complex infection but not with tuberculosis in AIDS patients. AB - The aim of the present study was to better characterize the expansion of double negative (DN) T cells in vivo in AIDS patients and to ascertain the discrepant response of an immunodepressed immune system towards two distinct mycobacterial infections. In a large cohort of HIV-1 seropositive patients with low CD4(+) T cell counts (<100/mm(3)), we have recently reported on an expansion of DN T cells which was observed only in patients with disseminated Mycobacterium avium infection, toxoplasmosis and Kaposi sarcoma, but not in patients with tuberculosis. The potential differential gammadelta T cells response observed in vivo in AIDS patients with tuberculosis or disseminated M. avium complex infection was investigated by collecting the concomitant or the closest T lymphocyte counts performed within 2 weeks of bacterial diagnosis of 112 disseminated M. avium infection and 41 tuberculosis patients. The DN and gammadelta T cell percentages were different between the two groups (P < 10(-4)) and the expansion of this compartment was found only with disseminated M. avium infections. An analysis of the variable delta2 segment versus pan-delta bearing T cells ratio disclosed a predominance of non-V(delta)2 T cells in these patients whose average values were identical in both groups. It is therefore concluded that the difference seen between these two types of mycobacterial infections concerning the DN T cells only involved the gammadelta T cells although the mechanism of their preferential expansion in disseminated M. avium infections remains a matter of speculation. PMID- 10464169 TI - Wortmannin inhibits translation of tumor necrosis factor-alpha in superantigen activated T cells. AB - The superantigen toxic shock syndrome toxin (TSST)-1 can induce tumor necrosis factor (TNF)-alpha expression in T cells and monocytes, through different signaling pathways. We have stimulated peripheral blood mononuclear cells with TSST-1 and found that the major cell producers of TNF-alpha as detected by cytofluorimetry and immunocytochemistry were CD4(+) T lymphocytes. The expression of TNF-alpha by CD4(+) T cells can be inhibited by either, wortmannin (WN) or LY 294002, two phosphatidylinositol 3-kinase (PI 3-K) inhibitors. The inhibitory effect is not transcriptional as WN does not change the mRNA steady state of TNF alpha at any of the concentrations tested and LY 294002 when preincubated with mononuclear cells at its median inhibitory concentration (IC(50) = 1. 4 microM) significantly inhibited the expression of TNF-alpha but not its mRNA. Immunoprecipitation of pulse-labeled intracellular TNF-alpha showed a specific decrease in the synthesis of this cytokine on cells treated with PI 3-K inhibitors. The p38 mitogen-activated protein kinase (MAPK) is involved in control of TNF-alpha translation in human macrophages. In T cells, we have found that the p38 MAPK inhibitor SB 203580 significantly decreased the secretion of TNF-alpha but not its mRNA. In addition, the combined use of WN and SB 203580 had an additive inhibitory effect on secretion of TNF-alpha. Therefore, both PI 3-K and p38 MAPK signaling pathways control TNF-alpha production in T cells. PMID- 10464171 TI - Functional diversity of natural IgM. AB - This paper proposes a method for the quantitative characterization of repertoire diversity of an unknown mixture of antibodies on the basis of its reactivity profile in the quantitative immunoblot (QIB). The QIB is calibrated by measuring the reactivity profiles of supernatants of known 'diversity' (i.e. known numbers of B cell clones). We define a quantitative 'index of variability' (IV) which decreases regularly as the diversity increases and the profiles tend towards a common 'convergence profile'. The calibration procedure is consolidated by a mathematical model based on the Poisson distribution; this theoretical model accounts correctly for the observed convergence behavior. On the basis of this calibration curve, it is possible to estimate the diversity of an unknown antibody mixture from a measure of its IV. We conclude that the functional diversity of natural serum IgM in mice can be estimated at approximately 16,000 clones. PMID- 10464170 TI - Immunopathogenesis of hepatic fibrosis in chronic liver injury induced by repeatedly administered concanavalin A. AB - Liver fibrosis is commonly observed in chronic liver disease. However, the immunological mechanisms underlying hepatic fibrosis due to chronic inflammation are not well defined, mainly because suitable experimental models have not been established. We have found that weekly i.v. administration of concanavalin A (Con A) in BALB/c mice brought about a striking alanine aminotransferase increase, resulting in piecemeal necrosis with bridging fibrosis in the parenchyma. Using this fibrosis model, we demonstrated the kinetics of cytokine mRNA expression in liver. Transforming growth factor (TGF)-beta1, TGF-alpha, basic fibroblast growth factor (bFGF) and hepatocyte growth factor mRNAs were up-regulated after each Con A administration. Furthermore, either anti-IFN-gamma, anti-tumor necrosis factor (TNF)-alpha or anti-TGF-beta mAb given together with Con A markedly inhibited the development of hepatic fibrosis. Treatment with either anti-IFN-gamma or anti-TNF alpha mAb also completely prevented hepatic injury; in contrast, treatment with anti-TGF-beta mAb did not. The treatment with anti-TGF-beta mAb did not affect the levels of hepatic mRNAs for either IFN-gamma or TNF-alpha after Con A injection. Treatment with either anti-IFN-gamma or anti-TNF-alpha did not affect the expression levels of TGF-beta in the liver. In conclusion, the continuous presence of both severe liver damage and up-regulation of TGF-beta synthesis is necessary to induce hepatic fibrosis in this model. PMID- 10464172 TI - Impaired thymopoietic potential of immature CD3(-)CD4(+)CD8(-) T cell precursors from SIV-infected rhesus monkeys. AB - Immature thymocyte subpopulations were examined for their capacity to differentiate in a newly developed xenogeneic monkey-mouse fetal thymus organ culture (FTOC) system. We provide evidence for impaired precursor function of CD3(-)CD4(+)CD8(-) thymocytes after in vivo infection with SIVmac251 as indicated by a reduced cell number per FTOC and a lower percentage of thymocytes with more mature phenotypes. Addition of recombinant SIV glycoprotein 120 (rgp120) also resulted in a dose-dependent impairment of T cell maturation in FTOC. The data suggest that in patients infected with HIV, T cell maturation and thus replenishment of peripheral pools may be compromised as a result of intrathymic infection or circulating viral gp120. PMID- 10464173 TI - Glucocorticoids increase the endocytic activity of human dendritic cells. AB - We have investigated the effect of glucocorticoids (GC) on antigen uptake molecule expression and on endocytic activity of human dendritic cells (DC). Human monocyte-derived DC were differentiated in vitro for 7 days with granulocyte macrophage colony stimulating factor and IL-4 in the presence or absence of dexamethasone 10(-8) M (Dex). Dex-treated DC showed an enhancement of mannose receptor (MR)-mediated endocytosis (measured as uptake of FITC-dextran) and of fluid-phase endocytosis [measured as uptake of Lucifer yellow (LY)] The effect was dose dependent and correlated with the length of exposure to Dex. The expression of receptors involved in antigen capture was investigated by FACS analysis. Dex up-regulates MR, CD16 and CD32 expression on DC. After maturation with tumor necrosis factor-alpha or CD40 ligand in Dex-treated DC, despite a reduction induced by maturation the endocytic activity of FITC-dextran and LY, the expression of MR, CD16 and CD32 remained higher than in control DC. In view of the fact that antigen capture was increased in cells cultured with Dex, we evaluated the ability to present soluble antigen that needs to be taken up and processed. Cells differentiated in the presence of Dex showed much lower efficiency in presenting tetanus toxin to specific autologous T cell lines. In conclusion our data suggest a new mechanism by which GC may influence immune responses. In fact with the increase in endocytic activity, Dex favors the scavenging of antigen from the external milieu, decreasing antigen concentration and availability, and simultaneously inhibiting the capacity to stimulate T cells. PMID- 10464174 TI - T cell interaction with ICAM-1-deficient endothelium in vitro: transendothelial migration of different T cell populations is mediated by endothelial ICAM-1 and ICAM-2. AB - The trafficking of T lymphocytes is carefully regulated by adhesive interactions with the vascular endothelium. Depending on their maturation and activation stage, T lymphocytes exhibit distinctive patterns of homing and recirculation, which is at least partly due to the selective expression of cell adhesion molecules (CAM) on the T cell surface. In order to define whether the differential usage of CAM during the steps of transendothelial migration is involved in organ-specific recirculation of different T cell subsets we compared the interaction of three different T cell populations with mouse endothelioma cell lines in vitro. Using a novel approach, where we directly compared T cell interaction with ICAM-1-deficient endothelium to wild-type endothelium, we recently demonstrated that endothelial ICAM-1 and ICAM-2 play a key role in mediating the transendothelial migration of CD4(+) memory T cells. Here we show that endothelial ICAM-1 and ICAM-2 are equally required for the transendothelial migration of other T cell populations such as thymocytes and T lymphoma cells, which differ from CD4(+) memory T cells in their maturation and activation stage, as well as in their surface expression of adhesion molecules. Our data therefore demonstrate that transendothelial migration of different T cell populations is mediated by the same endothelial CAM, i.e. ICAM-1 and ICAM-2, and thus subset specific interaction of T cells with endothelial cells must be regulated prior to transendothelial migration. PMID- 10464175 TI - Assessment of Cmv1 candidates by genetic mapping and in vivo antibody depletion of NK cell subsets. AB - The mouse chromosome 6 locus Cmv1 controls resistance to infection with murine cytomegalovirus (MCMV). We have previously shown that Cmv1 is tightly linked to members of the NK gene complex (NKC) including the Ly49 gene family. To assess the candidacy of individual NKC members for the resistance locus, first we followed the co-segregation of Cd94, Nkg2d, and the well-characterized Ly49a, Ly49c and Ly49g genes with respect to Cmv1 in pre-existing panels of intraspecific backcross mice. Gene order and intergene distances (in cM) were: centromere-Cd94/Nkg2d-(0.05)-Ly49a/Ly49c/Ly49 g/Cmv1-(0. 3) Prp/Kap/D6Mit13/111/219. This result excludes Cd94 and Nkg2d as candidates whereas it localizes the Ly49 genes within the minimal genetic interval for Cmv1. Second, we monitored the cell surface expression of individual Ly49 receptors in MCMV-infected mice over 2 weeks. The proportion of Ly49C(+) and Ly49C/I(+) cells decreased, the proportion of Ly49A(+) and Ly49G2(+) remained constant, and the cell surface density of Ly49G2 increased during infection, suggesting that NK cell subsets might have different roles in the regulation of MCMV infection. Third, we performed in vivo antibody depletion of specific NK cell subsets. Depletion with single antibodies did not affect the resistant phenotype suggesting that Ly49A(+), Ly49C(+), Ly49G2(+) and Ly49C/I(+) populations are not substantial players in MCMV resistance, and arguing for exclusion of the respective genes as candidates for Cmv1. In contrast, mice depleted with combined antibodies showed an intermediate phenotype. Whether residual NK cells, post depletion, belong to a particular subset expressing another Ly49 receptor, or a molecule encoded by a yet to be identified gene of the NKC, is discussed. PMID- 10464176 TI - T cell response in malaria pathogenesis: selective increase in T cells carrying the TCR V(beta)8 during experimental cerebral malaria. AB - To characterize the T cells involved in the pathogenesis of cerebral malaria (CM) induced by infection with Plasmodium berghei ANKA clone 1.49L (PbA 1.49L), the occurrence of the disease was assessed in mice lacking T cells of either the alphabeta or gammadelta lineage (TCRalphabeta(-/-) or TCRgammadelta(-/-)). TCRgammadelta(-/-) mice were susceptible to CM, whereas all TCRalphabeta(-/-) mice were resistant, suggesting that T cells of the alphabeta lineage are important in the genesis of CM. The repertoire of TCR V(beta) segment gene expression was examined by flow cytometry in B10.D2 mice, a strain highly susceptible to CM induced by infection with PbA 1.49L. In these mice, CM was associated with an increase of T cells bearing the V(beta)8.1, 2 segments in the peripheral blood lymphocytes. Most V(beta)8.1, 2(+) T cells from peripheral blood lymphocytes of the mice that developed CM belonged to the CD8 subset, and exhibited the CD69(+), CD44(high) and CD62L(low) phenotype surface markers. The link between the increase in V(beta)8.1, 2(+) T cells and the neuropathological consequences of PbA infection was strengthened by the observation that the occurrence of CM was significantly reduced in mice treated with KJ16 antibodies against the V(beta)8.1 and V(beta)8.2 chains, and in mice rendered deficient in V(beta)8.1(+) T cells by a mouse mammary tumor virus superantigen. PMID- 10464177 TI - Augmentation of CTLA-4 expression by wortmannin: involvement of lysosomal sorting properties of CTLA-4. AB - CTLA-4 (CD152) is transiently induced on the cell surface of activated T cells and expression is limited at a low level. In this study, we investigated the possibility that phosphatidylinositol 3 kinase (PI 3-K) and other related PI kinases associated with the cytoplasmic domain of CTLA-4 are involved in intracellular trafficking and sorting of CTLA-4 protein. Treatment with micromolar concentrations of wortmannin (WN) for >4 h enhanced both cell-surface and intracellular CTLA-4 without affecting its transcriptional activities in a murine mastocytoma cell line transfected with the human CTLA-4 gene and normal activated CD4(+) T cells. However, a more specific PI 3-K inhibitor, LY294002, failed to affect CTLA-4 expression, indicating that the action of WN is independent of conventional PI 3-K activities. WN down-regulated specific association of CTLA-4 with adaptor proteins and its endocytosis. The fact that lysosomotropic agents, ammonium chloride and monensin, enhanced CTLA-4 expression suggests that WN may also block lysosomal sorting and consequent degradation of CTLA-4. Co-localization of CTLA-4 and lysosome-associated membrane protein-1 detected by immunofluorescence microscopy indicates the actual lysosomal sorting of CTLA-4. Our data suggest the existence of WN-sensitive enzymes, which promote lysosomal sorting of CTLA-4. In addition to rapid endocytosis by clathrin associated adaptor complex, a prompt sorting of CTLA-4 to lysosomes may be one of the regulatory mechanisms for managing CTLA-4 signals in intracellular trafficking pathways. PMID- 10464179 TI - Unique CD4(+) T cells in TCR alpha chain-deficient class I MHC-restricted TCR transgenic mice: role in a superantigen-mediated disease process. AB - Mice carrying a transgenic TCR with targeted disruption of the TCR alpha chain (H Y alpha(-/-)) possess CD4(+) T cells which express the transgenic TCR beta without the alpha chain. These mice developed the murine acquired immunodeficiency syndrome (MAIDS) after infection with LP-BM5 retroviruses, a process which requires CD4(+) T cells. These cells are negative for TCR delta chain and pre-TCR alpha chain expression, and thus express a unique surface receptor with the TCR beta chain as a component. The cells respond to MAIDS virus associated superantigen and concanavalin A, but not to protein antigens such as ovalbumin. Thus, this novel surface receptor appears to play an important role in the pathogenesis of MAIDS. PMID- 10464178 TI - Genetic susceptibility or resistance to autoimmune encephalomyelitis in MHC congenic mice is associated with differential production of pro- and anti inflammatory cytokines. AB - Experimental allergic encephalomyelitis (EAE) is a T(h)1-type cell-mediated autoimmune disease induced by immunization with myelin proteins and mediated by CD4(+) T cells. Although susceptibility to EAE is dependent largely on MHC background, the B10.S strain is resistant to induction of EAE despite sharing the I-A(s) MHC locus with the susceptible SJL strain. Furthermore, NOD mice which spontaneously develop diabetes are susceptible to EAE induction with myelin oligodendrocyte glycoprotein (MOG) 35-55, whereas a MHC congenic strain, III, which also expresses I-A(g7) MHC haplotype does not develop diabetes and is also resistant to EAE induction. We induced EAE in these four strains of mice with MOG peptides 92-106 (for I-A(s) strains) and 35-55 (for I-A(g7) strains) in complete Freund's adjuvant. In the susceptible strains (SJL and NOD) in vitro, there are high levels of IFN-gamma production, whereas the resistant strains (B10.S or III) secreted primarily IL-4/IL-10 and transforming growth factor (TGF)-beta, and had decreased levels of IFN-gamma. When brains from susceptible and resistant mice were examined by immunohistochemical methods for cytokine expression, the brains from resistant mice showed fewer infiltrates which predominantly expressed IL-4 and IL-10 and/or TGF-beta. Brains from NOD and SJL with EAE showed mainly IL-2 and IFN-gamma positive cells. Thus, resistance to MOG induced EAE in B10.S and III mouse strains is related to non-MHC genes and is associated with an altered balance of pro- and anti-inflammatory cytokines both in lymphoid tissue and in the brain following immunization with myelin antigens. PMID- 10464180 TI - Bacterial DNA methylation: a cell cycle regulator? PMID- 10464181 TI - Evaluation of acyl coenzyme A oxidase (Aox) isozyme function in the n-alkane assimilating yeast Yarrowia lipolytica. AB - We have identified five acyl coenzyme A (CoA) oxidase isozymes (Aox1 through Aox5) in the n-alkane-assimilating yeast Yarrowia lipolytica, encoded by the POX1 through POX5 genes. The physiological function of these oxidases has been investigated by gene disruption. Single, double, triple, and quadruple disruptants were constructed. Global Aox activity was determined as a function of time after induction and of substrate chain length. Single null mutations did not affect growth but affected the chain length preference of acyl-CoA oxidase activity, as evidenced by a chain length specificity for Aox2 and Aox3. Aox2 was shown to be a long-chain acyl-CoA oxidase and Aox3 was found to be active against short-chain fatty acids, whereas Aox5 was active against molecules of all chain lengths. Mutations in Aox4 and Aox5 resulted in an increase in total Aox activity. The growth of mutant strains was analyzed. In the presence of POX1 only, strains did not grow on fatty acids, whereas POX4 alone elicited partial growth, and the growth of the double POX2-POX3-deleted mutant was normal excepted on plates containing oleic acid as the carbon source. The amounts of Aox protein detected by Western blotting paralleled the Aox activity levels, demonstrating the regulation of Aox in cells according to the POX genotype. PMID- 10464182 TI - Comparison of proteins involved in pilus synthesis and mating pair stabilization from the related plasmids F and R100-1: insights into the mechanism of conjugation. AB - F and R100-1 are closely related, derepressed, conjugative plasmids from the IncFI and IncFII incompatibility groups, respectively. Heteroduplex mapping and genetic analyses have revealed that the transfer regions are extremely similar between the two plasmids. Plasmid specificity can occur at the level of relaxosome formation, regulation, and surface exclusion between the two transfer systems. There are also differences in pilus serology, pilus-specific phage sensitivity, and requirements for OmpA and lipopolysaccharide components in the recipient cell. These phenotypic differences were exploited in this study to yield new information about the mechanism of pilus synthesis, mating pair stabilization, and surface and/or entry exclusion, which are collectively involved in mating pair formation (Mpf). The sequence of the remainder of the transfer region of R100-1 (trbA to traS) has been completed, and the complete sequence is compared to that of F. The differences between the two transfer regions include insertions and deletions, gene duplications, and mosaicism within genes, although the genes essential for Mpf are conserved in both plasmids. F+ cells carrying defined mutations in each of the Mpf genes were complemented with the homologous genes from R100-1. Our results indicate that the specificity in recipient cell recognition and entry exclusion are mediated by TraN and TraG, respectively, and not by the pilus. PMID- 10464183 TI - Combined genetic and physical map of the complex genome of Agrobacterium tumefaciens. AB - A combined genetic and physical map of the Agrobacterium tumefaciens A348 (derivative of C58) genome was constructed to address the discrepancy between initial single-chromosome genetic maps and more recent physical mapping data supporting the presence of two nonhomologous chromosomes. The combined map confirms the two-chromosome genomic structure and the correspondence of the initial genetic maps to the circular chromosome. The linear chromosome is almost devoid of auxotrophic markers, which probably explains why it was missed by genetic mapping studies. PMID- 10464184 TI - Timing of FtsZ assembly in Escherichia coli. AB - The timing of the appearance of the FtsZ ring at the future site of division in Escherichia coli was determined by in situ immunofluorescence microscopy for two strains grown under steady-state conditions. The strains, B/rA and K-12 MC4100, differ largely in the duration of the D period, the time between termination of DNA replication and cell division. In both strains and under various growth conditions, the assembly of the FtsZ ring was initiated approximately simultaneously with the start of the D period. This is well before nucleoid separation or initiation of constriction as determined by fluorescence and phase contrast microscopy. The durations of the Z-ring period, the D period, and the period with a visible constriction seem to be correlated under all investigated growth conditions in these strains. These results suggest that (near) termination of DNA replication could provide a signal that initiates the process of cell division. PMID- 10464185 TI - Molecular characterization of type-specific capsular polysaccharide biosynthesis genes of Streptococcus agalactiae type Ia. AB - The type-specific capsular polysaccharide (CP) of a group B streptococcus, Streptococcus agalactiae type Ia, is a high-molecular-weight polymer consisting of the pentasaccharide repeating unit 4)-[alpha-D-NeupNAc-(2-->3)-beta-D-Galp-(1- >4)-beta-D-GlcpNAc-(1- ->3 )]-beta-D-Galp-(1-->4)-beta-D-Glcp-(1. Here, cloning, sequencing, and transcription of the type Ia-specific capsular polysaccharide synthesis (cps) genes and functional analysis of these gene products are described. A 26-kb DNA fragment containing 18 complete open reading frames (ORFs) was cloned. These ORFs were designated cpsIaA to cpsIaL, neu (neuraminic acid synthesis gene) A to D, orf1 and ung (uracil DNA glycosylase). The cps gene products of S. agalactiae type Ia were homologous to proteins involved in CP synthesis of S. agalactiae type III and S. pneumoniae serotype 14. Unlike the cps gene cluster of S. pneumoniae serotype 14, transcription of this operon may start from cpsIaA, cpsIaE, and orf1 because putative promoter sequences were found in front of these genes. Northern hybridization, reverse transcription-PCR, and primer extension analyses supported this hypothesis. DNA sequence analysis showed that there were two transcriptional terminators in the 3' end of this operon (downstream of orf1 and ung). The functions of CpsIaE, CpsIaG, CpsIaI, and CpsIaJ were examined by glycosyltransferase assay by using the gene products expressed in Escherichia coli JM109 harboring plasmids containing various S. agalactiae type Ia cps gene fragments. Enzyme assays suggested that the gene products of cpsIaE, cpsIaG, cpsIaI, and cpsIaJ are putative glucosyltransferase, beta-1, 4 galactosyltransferase, beta-1,3-N-acetylglucosaminyltransferase, and beta-1,4 galactosyltransferase, respectively. PMID- 10464186 TI - Amino acid-DNA contacts by RhaS: an AraC family transcription activator. AB - RhaS, an AraC family protein, activates rhaBAD transcription by binding to rhaI, a site consisting of two 17-bp inverted repeat half-sites. In this work, amino acids in RhaS that make base-specific contacts with rhaI were identified. Sequence similarity with AraC suggested that the first contacting motif of RhaS was a helix-turn-helix. Assays of rhaB-lacZ activation by alanine mutants within this potential motif indicated that residues 201, 202, 205, and 206 might contact rhaI. The second motif was identified based on the hypothesis that a region of especially high amino acid similarity between RhaS and RhaR (another AraC family member) might contact the nearly identical DNA sequences in one major groove of their half-sites. We first made targeted, random mutations and then made alanine substitutions within this region of RhaS. Our analysis identified residues 247, 248, 250, 252, 253, and 254 as potentially important for DNA binding. A genetic loss-of-contact approach was used to identify whether any of the RhaS amino acids in the first or second contacting motif make base-specific DNA contacts. In motif 1, we found that Arg202 and Arg206 both make specific contacts with bp -65 and 67 in rhaI1, and that Arg202 contacts -46 and Arg206 contacts -48 in rhaI2. In motif 2, we found that Asp250 and Asn252 both contact the bp -79 in rhaI1. Alignment with the recently crystallized MarA protein suggest that both RhaS motifs are likely helix-turn-helix DNA-binding motifs. PMID- 10464187 TI - An autoregulatory circuit affecting peptide signaling in Bacillus subtilis. AB - The competence and sporulation factor (CSF) of Bacillus subtilis is an extracellular pentapeptide produced from the product of phrC. CSF has at least three activities: (i) at low concentrations, it stimulates expression of genes activated by the transcription factor ComA; at higher concentrations, it (ii) inhibits expression of those same genes and (iii) stimulates sporulation. Because the activities of CSF are concentration dependent, we measured the amount of extracellular CSF produced by cells. We found that by mid-exponential phase, CSF accumulated to concentrations (1 to 5 nM) that stimulate ComA-dependent gene expression. Upon entry into stationary phase, CSF reached 50 to 100 nM, concentrations that stimulate sporulation and inhibit ComA-dependent gene expression. Transcription of phrC was found to be controlled by two promoters: P1, which precedes rapC, the gene upstream of phrC; and P2, which directs transcription of phrC only. Both RapC and CSF were found to be part of autoregulatory loops that affect transcription from P1, which we show is activated by ComA approximately P. RapC negatively regulates its own expression, presumably due to its ability to inhibit accumulation of ComA approximately P. CSF positively regulates its own expression, presumably due to its ability to inhibit RapC activity. Transcription from P2, which is controlled by the alternate sigma factor sigma(H), increased as cells entered stationary phase, contributing to the increase in extracellular CSF at this time. In addition to controlling transcription of phrC, sigmaH appears to control expression of at least one other gene required for production of CSF. PMID- 10464188 TI - Distinct types of rRNA operons exist in the genome of the actinomycete Thermomonospora chromogena and evidence for horizontal transfer of an entire rRNA operon. AB - We describe here the presence of two distinct types of rRNA operons in the genome of a thermophilic actinomycete Thermomonospora chromogena. The genome of T. chromogena contains six rRNA operons (rrn), of which four complete and two incomplete ones were cloned and sequenced. Comparative analysis revealed that the operon rrnB exhibits high levels of sequence variations to the other five nearly identical ones throughout the entire length of the operon. The coding sequences for the 16S and 23S rRNA genes differ by approximately 6 and 10%, respectively, between the two types of operons. Normal functionality of rrnB is concluded on the basis of the nonrandom distribution of nucleotide substitutions, the presence of compensating nucleotide covariations, the preservation of secondary and tertiary rRNA structures, and the detection of correctly processed rRNAs in the cell. Comparative sequence analysis also revealed a close evolutionary relationship between rrnB operon of T. chromogena and rrnA operon of another thermophilic actinomycete Thermobispora bispora. We propose that T. chromogena acquired rrnB operon from T. bispora or a related organism via horizontal gene transfer. PMID- 10464189 TI - Effect of ionic strength on initial interactions of Escherichia coli with surfaces, studied on-line by a novel quartz crystal microbalance technique. AB - A novel quartz crystal microbalance (QCM) technique was used to study the adhesion of nonfimbriated and fimbriated Escherichia coli mutant strains to hydrophilic and hydrophobic surfaces at different ionic strengths. This technique enabled us to measure both frequency shifts (Deltaf), i.e., the increase in mass on the surface, and dissipation shifts (DeltaD), i.e., the viscoelastic energy losses on the surface. Changes in the parameters measured by the extended QCM technique reflect the dynamic character of the adhesion process. We were able to show clear differences in the viscoelastic behavior of fimbriated and nonfimbriated cells attached to surfaces. The interactions between bacterial cells and quartz crystal surfaces at various ionic strengths followed different trends, depending on the cell surface structures in direct contact with the surface. While Deltaf and DeltaD per attached cell increased for nonfimbriated cells with increasing ionic strengths (particularly on hydrophobic surfaces), the adhesion of the fimbriated strain caused only low-level frequency and dissipation shifts on both kinds of surfaces at all ionic strengths tested. We propose that nonfimbriated cells may get better contact with increasing ionic strengths due to an increased area of contact between the cell and the surface, whereas fimbriated cells seem to have a flexible contact with the surface at all ionic strengths tested. The area of contact between fimbriated cells and the surface does not increase with increasing ionic strengths, but on hydrophobic surfaces each contact point seems to contribute relatively more to the total energy loss. Independent of ionic strength, attached cells undergo time-dependent interactions with the surface leading to increased contact area and viscoelastic losses per cell, which may be due to the establishment of a more intimate contact between the cell and the surface. Hence, the extended QCM technique provides new qualitative information about the direct contact of bacterial cells to surfaces and the adhesion mechanisms involved. PMID- 10464190 TI - The dual-specificity protein phosphatase Yvh1p acts upstream of the protein kinase mck1p in promoting spore development in Saccharomyces cerevisiae. AB - Diploid Saccharomyces cerevisiae cells induce YVH1 expression and enter the developmental pathway, leading to sporulation when starved for nitrogen. We show that yvh1 disruption causes a defect in spore maturation; overexpression of MCK1 or IME1 suppresses this yvh1 phenotype. While mck1 mutations are epistatic to those in yvh1 relative to spore maturation, overexpression of MCK1 does not suppress the yvh1 slow-vegetative-growth phenotype. We conclude that (i) Yvh1p functions earlier than Mck1p and Ime1p in the signal transduction cascade that regulates sporulation and is triggered by nitrogen starvation and (ii) the role of Yvh1p in gametogenesis can be genetically distinguished from its role in vegetative growth. PMID- 10464191 TI - Translation of two nested genes in bacteriophage P4 controls immunity-specific transcription termination. AB - In phage P4, transcription of the left operon may occur from both the constitutive PLE promoter and the regulated PLL promoter, about 400 nucleotides upstream of PLE. A strong Rho-dependent termination site, timm, is located downstream of both promoters. When P4 immunity is expressed, transcription starting at PLE is efficiently terminated at timm, whereas transcription from PLL is immunity insensitive and reads through timm. We report the identification of two nested genes, kil and eta, located in the P4 left operon. The P4 kil gene, which encodes a 65-amino-acid polypeptide, is the first translated gene downstream of the PLE promoter, and its expression is controlled by P4 immunity. Overexpression of kil causes cell killing. This gene is the terminal part of a longer open reading frame, eta, which begins upstream of PLE. The eta gene is expressed when transcription starts from the PLL promoter. Three likely start codons predict a size between 197 and 199 amino acids for the Eta gene product. Both kil and eta overlap the timm site. By cloning kil upstream of a tRNA reporter gene, we demonstrated that translation of the kil region prevents premature transcription termination at timm. This suggests that P4 immunity might negatively control kil translation, thus enabling transcription termination at timm. Transcription starting from PL proceeds through timm. Mutations that create nonsense codons in eta caused premature termination of transcription starting from PLL. Suppression of the nonsense mutation restored transcription readthrough at timm. Thus, termination of transcription from PLL is prevented by translation of eta. PMID- 10464192 TI - Analysis of BvgA activation of the pertactin gene promoter in Bordetella pertussis. AB - Bordetella pertussis, the causative agent of whooping cough, regulates expression of its virulence factors via a two-component signal transduction system encoded by the bvg regulatory locus. It has been shown by activation kinetics that several of the virulence factors are differentially regulated. fha is transcribed at 10 min following an inducing signal, while ptx is not transcribed until 2 to 4 h after the inducing signal. We present data indicating that prn is transcribed at 1 h, an intermediate time compared to those of fha and ptx. We have identified cis-acting sequences necessary for expression of prn in B. pertussis by using prn lac fusions containing alterations in the sequence upstream of the prn open reading frame. In vitro transcription and DNase I footprinting analyses provided evidence to support our hypothesis that BvgA binds to this sequence upstream of prn to activate transcription from the promoter. Our genetic data indicate that the region critical for prn activation extends upstream to position -84. However, these data do not support the location of the prn transcription start site as previously published. We used a number of methods, including prn-lac fusions, reverse transcriptase PCR, and 5' rapid amplification of cDNA ends, to localize and identify the bvg-dependent 5' end of the prn transcript to the cytosine at 125 with respect to the published start site. PMID- 10464193 TI - 16S rRNA is bound to era of Streptococcus pneumoniae. AB - Era is an essential membrane-associated GTPase that is present in bacteria and mycoplasmas. Era appears to play an important role in the regulation of the bacterial cell cycle. In this study, we expressed the native and glutathione S transferase (GST) fusion forms of Streptococcus pneumoniae Era in Escherichia coli and purified both proteins to homogeneity. We showed that RNA was copurified with the GST-Era protein of S. pneumoniae during affinity purification and remained associated with the protein after removal of the GST tag by thrombin cleavage. The thrombin-treated and untreated GST-Era proteins could bind and hydrolyze GTP and exhibited similar kinetic properties (dissociation constant [kD], Km, and Vmax). However, the native Era protein purified by using different chromatographic columns had a much lower GTPase activity than did GST-Era, although it had a similar k(D). In addition, RNA was not associated with the protein. Purified GST-Era protein was shown to be present as high (600-kDa)- and low (120-kDa)-molecular-mass forms. The high-molecular-mass form of GST-Era was associated with RNA and exhibited a very high GTPase activity. Approximately 40% of purified GST-Era protein was associated with RNA, and removal of the RNA resulted in a significant reduction in GTPase activity. The RNA associated with GST-Era was shown to be predominantly 16S rRNA. The native Era protein isolated directly from S. pneumoniae was also present as a high-molecular-mass species (600 kDa) complexed with RNA. Together, our results suggest that 16S rRNA is associated with Era and might stimulate its GTPase activity. PMID- 10464194 TI - Response of hya expression to external pH in Escherichia coli. AB - The hya operon of Escherichia coli is composed of the genes which synthesize uptake hydrogenase isoenzyme 1 (Hyd1). Although hya expression and Hyd1 synthesis occur only under anaerobic conditions, Hyd1 is not essential for growth. In this study we used a hya'-'lacZ fusion to characterize parameters of anaerobic growth that maximize hya expression in an attempt to further elucidate Hyd1 function. We found that the expression pattern of hya followed a decline of external pH. In buffered media where the pH value was set, the onset of hya expression initiated earlier in growth and reached a greater peak level in acidic than in alkaline medium. When cultures expressing hya were shifted from acidic to alkaline conditions, hya expression was arrested; shifting from alkaline to acidic conditions stimulated hya expression. Maximal expression of hya under all growth conditions required the sigma factor RpoS and transcriptional regulators AppY and ArcA. In the absence of RpoS or AppY, the response of hya expression onset to external pH was evident and maximal hya levels remained greater in acidic than in alkaline medium. However, the absence of ArcA led to a diminished response of expression onset to external pH and the loss of elevated expression at an acidic external pH. The fermentation end product formate slightly altered hya expression levels but was not required for hya to respond to external pH. In contrast to hya expression, the onset of hyb operon expression, encoding uptake hydrogenase isoenzyme 2, was constitutive with respect to external pH. However, external pH did affect hyb expression levels, which, in contrast to hya, were maximal in alkaline rather than acidic medium. PMID- 10464195 TI - Suppression of nonsense mutations induced by expression of an RNA complementary to a conserved segment of 23S rRNA. AB - We identified a short RNA fragment, complementary to the Escherichia coli 23S rRNA segment comprising nucleotides 735 to 766 (in domain II), which when expressed in vivo results in the suppression of UGA nonsense mutations in two reporter genes. Neither UAA nor UAG mutations, examined at the same codon positions, were suppressed by the expression of this antisense rRNA fragment. Our results suggest that a stable phylogenetically conserved hairpin at nucleotides 736 to 760 in 23S rRNA, which is situated close to the peptidyl transferase center, may participate in one or more specific interactions during peptide chain termination. PMID- 10464197 TI - HWP1 functions in the morphological development of Candida albicans downstream of EFG1, TUP1, and RBF1. AB - The morphological plasticity of Candida albicans is an important determinant of pathogenicity, and nonfilamentous mutants are avirulent. HWP1, a hypha-specific gene, was identified in a genetic screen for developmentally regulated genes and encodes a cell surface protein of unknown function. Heterozygous and homozygous deletions of HWP1 resulted in a medium-conditional defect in hyphal development. HWP1 expression was blocked in a Deltaefg1 mutant, reduced in an Deltarbf1 mutant, and derepressed in a Deltatup1 mutant. Therefore, HWP1 functions downstream of the developmental regulators EFG1, TUP1, and RBF1. Mutation of CPH1 had no effect on HWP1 expression, suggesting that the positive regulators of hyphal development, CPH1 and EFG1, are components of separate pathways with different target genes. The expression of a second developmentally regulated gene, ECE1, was similarly regulated by EFG1. Since ECE1 is not required for hyphal development, the regulatory role of EFG1 apparently extends beyond the control of cell shape determinants. However, expression of ECE1 was not influenced by TUP1, suggesting that there may be some specificity in the regulation of morphogenic elements during hyphal development. PMID- 10464196 TI - The Cpx envelope stress response is controlled by amplification and feedback inhibition. AB - In Escherichia coli, the Cpx two-component regulatory system activates expression of protein folding and degrading factors in response to misfolded proteins in the bacterial envelope (inner membrane, periplasm, and outer membrane). It is comprised of the histidine kinase CpxA and the response regulator CpxR. This response plays a role in protection from stresses, such as elevated pH, as well as in the biogenesis of virulence factors. Here, we show that the Cpx periplasmic stress response is subject to amplification and repression through positive and negative autofeedback mechanisms. Western blot and operon fusion analyses demonstrated that the cpxRA operon is autoactivated. Conditions that lead to elevated levels of phosphorylated CpxR cause a concomitant increase in transcription of cpxRA. Conversely, overproduction of CpxP, a small, Cpx regulated protein of previously unknown function, represses the regulon and can block activation of the pathway. This repression is dependent on an intact CpxA sensing domain. The ability to autoactivate and then subsequently repress allows for a temporary amplification of the Cpx response that may be important in rescuing cells from transitory stresses and cueing the appropriately timed elaboration of virulence factors. PMID- 10464198 TI - Reduction of adenosine-5'-phosphosulfate instead of 3'-phosphoadenosine-5' phosphosulfate in cysteine biosynthesis by Rhizobium meliloti and other members of the family Rhizobiaceae. AB - We have cloned and sequenced three genes from Rhizobium meliloti (Sinorhizobium meliloti) that are involved in sulfate activation for cysteine biosynthesis. Two of the genes display homology to the Escherichia coli cysDN genes, which code for an ATP sulfurylase (EC 2.7.7.4). The third gene has homology to the E. coli cysH gene, a 3'-phosphoadenosine-5'-phosphosulfate (PAPS) reductase (EC 1.8.99.4), but has greater homology to a set of genes found in Arabidopsis thaliana that encode an adenosine-5'-phosphosulfate (APS) reductase. In order to determine the specificity of the R. meliloti reductase, the R. meliloti cysH homolog was histidine tagged and purified, and its specificity was assayed in vitro. Like the A. thaliana reductases, the histidine-tagged R. meliloti cysH gene product appears to favor APS over PAPS as a substrate, with a Km for APS of 3 to 4 microM but a Km for PAPS of >100 microM. In order to determine whether this preference for APS is unique to R. meliloti among members of the family Rhizobiaceae or is more widespread, cell extracts from R. leguminosarum, Rhizobium sp. strain NGR234, Rhizobium fredii (Sinorhizobium fredii), and Agrobacterium tumefaciens were assayed for APS or PAPS reductase activity. Cell extracts from all four species also preferentially reduce APS over PAPS. PMID- 10464199 TI - Cloning and sequence analysis of a new cellulase gene encoding CelK, a major cellulosome component of Clostridium thermocellum: evidence for gene duplication and recombination. AB - The cellulolytic and hemicellulolytic complex of Clostridium thermocellum, termed cellulosome, consists of up to 26 polypeptides, of which at least 17 have been sequenced. They include 12 cellulases, 3 xylanases, 1 lichenase, and CipA, a scaffolding polypeptide. We report here a new cellulase gene, celK, coding for CelK, a 98-kDa major component of the cellulosome. The gene has an open reading frame (ORF) of 2,685 nucleotides coding for a polypeptide of 895 amino acid residues with a calculated mass of 100,552 Da. A signal peptide of 27 amino acid residues is cut off during secretion, resulting in a mature enzyme of 97,572 Da. The nucleotide sequence is highly similar to that of cbhA (V. V. Zverlov et al., J. Bacteriol. 180:3091-3099, 1998), having an ORF of 3,690 bp coding for the 1,230-amino-acid-residue CbhA of the same bacterium. Homologous regions of the two genes are 86.5 and 84.3% identical without deletion or insertion on the nucleotide and amino acid levels, respectively. Both have domain structures consisting of a signal peptide, a family IV cellulose binding domain (CBD), a family 9 glycosyl hydrolase domain, and a dockerin domain. A striking distinction between the two polypeptides is that there is a 330-amino-acid insertion in CbhA between the catalytic domain and the dockerin domain containing a fibronectin type 3-like domain and family III CBD. This insertion, missing in CelK, is responsible for the size difference between CelK and CbhA. Upstream and downstream flanking sequences of the two genes show no homology. The data indicate that celK and cbhA in the genome of C. thermocellum have evolved through gene duplication and recombination of domain coding sequences. celK without a dockerin domain was expressed in Escherichia coli and purified. The enzyme had pH and temperature optima at 6.0 and 65 degrees C, respectively. It hydrolyzed p nitrophenyl-beta-D-cellobioside with a Km and a Vmax of 1.67 microM and 15.1 U/mg, respectively. Cellobiose was a strong inhibitor of CelK activity, with a Ki of 0.29 mM. The enzyme was thermostable, after 200 h of incubation at 60 degrees C, 97% of the original activity remained. Properties of the enzyme indicated that it is a cellobiohydrolase. PMID- 10464200 TI - A- and T-tract-mediated intrinsic curvature in native DNA between the binding site of the upstream activator NtrC and the nifLA promoter of Klebsiella pneumoniae facilitates transcription. AB - The nif promoters of Klebsiella pneumoniae must be activated by proteins bound to upstream sequences which are thought to interact with the sigma54-RNA polymerase holoenzyme by DNA looping. NifA is the activator for most of the promoters, and integration host factor (IHF) mediates the DNA looping. While NtrC is the activator for the nifLA promoter, no IHF appears to be involved. There are two A tracts and one T tract between the upstream enhancer and the nifLA promoter. This DNA segment exhibits anomalous electrophoretic mobility, suggesting intrinsic sequence-induced curvature in the DNA. On the one hand, mutation of the A tracts or T tract individually or together, or deletion of the A tracts and the T tract reduces the anomaly; on the other hand, creation of two additional A tracts enhances the anomaly. Intrinsic curvature in the DNA has been confirmed by circular permutation analysis after cloning the DNA fragment in the vector pBend 2 and also by electron microscopy. Computer simulation with the DNA base sequence is also suggestive of intrinsic curvature. A transcriptional fusion with the Escherichia coli lacZ gene of the DNA fragment containing the nifLA promoter and the wild-type or the mutated upstream sequences was constructed, and in vivo transcription in K. pneumoniae and E. coli was monitored. There was indeed very good correlation between the extent of intrinsic curvature of the DNA and transcription from the promoter, suggesting that DNA curvature due to the A tracts and the T tract was necessary for transcription in vivo from the nifLA promoter of K. pneumoniae. PMID- 10464201 TI - The napF and narG nitrate reductase operons in Escherichia coli are differentially expressed in response to submicromolar concentrations of nitrate but not nitrite. AB - Escherichia coli synthesizes two biochemically distinct nitrate reductase enzymes, a membrane-bound enzyme encoded by the narGHJI operon and a periplasmic cytochrome c-linked nitrate reductase encoded by the napFDAGHBC operon. To address why the cell makes these two enzymes, continuous cell culture techniques were used to examine napF and narG gene expression in response to different concentrations of nitrate and/or nitrite. Expression of the napF-lacZ and narG lacZ reporter fusions in strains grown at different steady-state levels of nitrate revealed that the two nitrate reductase operons are differentially expressed in a complementary pattern. The napF operon apparently encodes a "low substrate-induced" reductase that is maximally expressed only at low levels of nitrate. Expression is suppressed under high-nitrate conditions. In contrast, the narGHJI operon is only weakly expressed at low nitrate levels but is maximally expressed when nitrate is elevated. The narGHJI operon is therefore a "high substrate-induced" operon that somehow provides a second and distinct role in nitrate metabolism by the cell. Interestingly, nitrite, the end product of each enzyme, had only a minor effect on the expression of either operon. Finally, nitrate, but not nitrite, was essential for repression of napF gene expression. These studies reveal that nitrate rather than nitrite is the primary signal that controls the expression of these two nitrate reductase operons in a differential and complementary fashion. In light of these findings, prior models for the roles of nitrate and nitrite in control of narG and napF expression must be reconsidered. PMID- 10464202 TI - Signal-dependent phosphorylation of the membrane-bound NarX two-component sensor transmitter protein of Escherichia coli: nitrate elicits a superior anion ligand response compared to nitrite. AB - The Nar two-component regulatory system, consisting of the dual sensor transmitters NarX and NarQ and the dual response regulators NarL and NarP, controls the expression of various anaerobic respiratory pathway genes and fermentation pathway genes. Although both NarX and NarQ are known to detect the two environmental signals nitrate and nitrite, little is known regarding the sensitivity and selectivity of ligand for detection or activation of the sensor transmitters. In this study, we have developed a sensitive anion-specific in vitro assay for NarX autophosphorylation by using Escherichia coli membranes highly enriched in the full-length NarX protein. In this ATP- and magnesium dependent reaction, nitrate elicited a greater signal output (i.e., NarX autophosphorylation) than did nitrite. Nitrate stimulation occurred at concentrations as low as 5 microM, and the half-maximal level of NarX autophosphorylation occurred at approximately 35 microM nitrate. In contrast, nitrite-dependent stimulation was detected only at 500 microM, while 3.5 mM nitrite was needed to achieve half-maximal NarX autophosphorylation. Maximal nitrate- and nitrite-stimulated levels of NarX phosphorylation were five and two times, respectively, over the basal level of NarX autophosphorylation. The presence of Triton X-100 eliminated the nitrate-stimulated kinase activity and lowered the basal level of activity, suggesting that the membrane environment plays a crucial role in nitrate detection and/or regulation of kinase activity. These results provide in vitro evidence for the differential detection of dual signaling ligands by the NarX sensor-transmitter protein, which modulates the cytoplasmic NarX autokinase activity and phosphotransfer to NarL, the cognate response regulator. PMID- 10464203 TI - The 17-gene ethanolamine (eut) operon of Salmonella typhimurium encodes five homologues of carboxysome shell proteins. AB - The eut operon of Salmonella typhimurium encodes proteins involved in the cobalamin-dependent degradation of ethanolamine. Previous genetic analysis revealed six eut genes that are needed for aerobic use of ethanolamine; one (eutR), encodes a positive regulator which mediates induction of the operon by vitamin B12 plus ethanolamine. The DNA sequence of the eut operon included 17 genes, suggesting a more complex pathway than that revealed genetically. We have correlated an open reading frame in the sequence with each of the previously identified genes. Nonpolar insertion and deletion mutations made with the Tn10 derived transposable element T-POP showed that at least 10 of the 11 previously undetected eut genes have no Eut phenotype under the conditions tested. Of the dispensable eut genes, five encode apparent homologues of proteins that serve (in other organisms) as shell proteins of the carboxysome. This bacterial organelle, found in photosynthetic and sulfur-oxidizing bacteria, may contribute to CO2 fixation by concentrating CO2 and excluding oxygen. The presence of these homologues in the eut operon of Salmonella suggests that CO2 fixation may be a feature of ethanolamine catabolism in Salmonella. PMID- 10464205 TI - Cold shock response of Bacillus subtilis: isoleucine-dependent switch in the fatty acid branching pattern for membrane adaptation to low temperatures. AB - Bacillus subtilis has developed sophisticated mechanisms to withstand fluctuations in temperature. Membrane fatty acids are the major determinants for a sufficiently fluid membrane state to ensure the membrane's function at all temperatures. The fatty acid profile of B. subtilis is characterized by a high content of branched fatty acids irrespective of the growth medium. Here, we report on the importance of isoleucine for B. subtilis to survive cold shock from 37 to 15 degrees C. Cold shock experiments with strain JH642 revealed a cold protective function for all intermediates of anteiso-branched fatty acid biosynthesis. Metabolites related to iso-branched or straight-chain fatty acid biosynthesis were not protective. Fatty acid profiles of different B. subtilis wild-type strains proved the altered branching pattern by an increase in the anteiso-branched fatty acid content and a concomitant decrease of iso-branched species during cold shock. There were no significant changes in the fatty acid saturation or acyl chain length. The cold-sensitive phenotype of isoleucine deficient strains in the absence of isoleucine correlated with their inability to synthesize more anteiso-branched fatty acids, as shown by the fatty acid profile. The switch to a fatty acid profile dominated by anteiso-C(15:0) and C(17:0) at low temperatures and the cold-sensitive phenotype of isoleucine-deficient strains in the absence of isoleucine focused our attention on the critical role of anteiso-branched fatty acids in the growth of B. subtilis in the cold. PMID- 10464204 TI - Identification of a two-component signal transduction system from Corynebacterium diphtheriae that activates gene expression in response to the presence of heme and hemoglobin. AB - Corynebacterium diphtheriae, the causative agent of diphtheria, utilizes various host compounds to acquire iron. The C. diphtheriae hmuO gene encodes a heme oxygenase that is involved in the utilization of heme and hemoglobin as iron sources. Transcription of the hmuO gene in C. diphtheriae is controlled under a dual regulatory mechanism in which the diphtheria toxin repressor protein (DtxR) and iron repress expression while either heme or hemoglobin is needed to activate transcription. In this study, two clones isolated from a C. diphtheriae chromosomal library were shown to activate transcription from the hmuO promoter in Escherichia coli. Sequence analysis revealed that these activator clones each carried distinct genes whose products had significant homology to response regulators of two-component signal transduction systems. Located upstream from each of these response regulator homologs are partial open reading frames that are predicted to encode the C-terminal portions of sensor kinases. The full length sensor kinase gene for each of these systems was cloned from the C. diphtheriae chromosome, and constructs each carrying one complete sensor kinase gene and its cognate response regulator were constructed. One of these constructs, pTSB20, which carried the response regulator (chrA) and its cognate sensor kinase (chrS), was shown to strongly activate transcription from the hmuO promoter in a heme-dependent manner in E. coli. A mutation in chrA (chrAD50N), which changed a conserved aspartic acid residue at position 50, the presumed site of phosphorylation by ChrS, to an asparagine, abolished heme-dependent activation. These findings suggest that the sensor kinase ChrS is involved in the detection of heme and the transduction of this signal, via a phosphotransfer mechanism, to the response regulator ChrA, which then activates transcription of the hmuO promoter. This is the first report of a bacterial two-component signal transduction system that controls gene expression through a heme-responsive mechanism. PMID- 10464206 TI - Direct probing of the surface ultrastructure and molecular interactions of dormant and germinating spores of Phanerochaete chrysosporium. AB - Atomic force microscopy (AFM) has been used to probe, under physiological conditions, the surface ultrastructure and molecular interactions of spores of the filamentous fungus Phanerochaete chrysosporium. High-resolution images revealed that the surface of dormant spores was uniformly covered with rodlets having a periodicity of 10 +/- 1 nm, which is in agreement with earlier freeze etching measurements. In contrast, germinating spores had a very smooth surface partially covered with rough granular structures. Force-distance curve measurements demonstrated that the changes in spore surface ultrastructure during germination are correlated with profound modifications of molecular interactions: while dormant spores showed no adhesion with the AFM probe, germinating spores exhibited strong adhesion forces, of 9 +/- 2 nN magnitude. These forces are attributed to polysaccharide binding and suggested to be responsible for spore aggregation. This study represents the first direct characterization of the surface ultrastructure and molecular interactions of living fungal spores at the nanometer scale and offers new prospects for mapping microbial cell surface properties under native conditions. PMID- 10464207 TI - Comparative genetics of capsular polysaccharide biosynthesis in Streptococcus pneumoniae types belonging to serogroup 19. AB - The genetic basis for the structural diversity of capsule polysaccharide (CPS) in Streptococcus pneumoniae serogroup 19 (consisting of types 19F, 19A, 19B, and 19C) has been determined for the first time. In this study, the genetic basis for the 19A and 19C serotypes is described, and the structures of all four serogroup 19 cps loci and their flanking sequences are compared. Transformation studies show that the structural difference between the 19A and 19F CPSs is likely to be a consequence of differences between their respective polysaccharide polymerase genes (cps19aI and cps19fI). The CPS of type 19C differs from that of type 19B by the addition of glucose. We have identified a single gene difference between the two cps loci (cps19cS), which is likely to encode a glucosyl transferase. The arrangement of the genes within the cps19 loci is highly conserved, with 13 genes (cps19A to -H and cps19K to -O) common to all four serogroup 19 members. These cps genes encode functions required for the synthesis of the shared trisaccharide component of the group 19 CPS repeat unit structures. Furthermore, the genetic differences between the group 19 cps loci identified are consistent with the CPS structures of the individual serotypes. Functions have been assigned to nearly all of the cps19 gene products, based on either gene complementation or similarity to other proteins with known functions, and putative biosynthetic pathways for production of all four group 19 CPSs have been proposed. PMID- 10464208 TI - An engineered cytochrome b6c1 complex with a split cytochrome b is able to support photosynthetic growth of Rhodobacter capsulatus. AB - The ubihydroquinone-cytochrome c oxidoreductase (or the cytochrome bc1 complex) from Rhodobacter capsulatus is composed of the Fe-S protein, cytochrome b, and cytochrome c1 subunits encoded by petA(fbcF), petB(fbcB), and petC(fbcC) genes organized as an operon. In the work reported here, petB(fbcB) was split genetically into two cistrons, petB6 and petBIV, which encoded two polypeptides corresponding to the four amino-terminal and four carboxyl-terminal transmembrane helices of cytochrome b, respectively. These polypeptides resembled the cytochrome b6 and su IV subunits of chloroplast cytochrome b6f complexes, and together with the unmodified subunits of the cytochrome bc1 complex, they formed a novel enzyme, named cytochrome b6c1 complex. This membrane-bound multisubunit complex was functional, and despite its smaller amount, it was able to support the photosynthetic growth of R. capsulatus. Upon further mutagenesis, a mutant overproducing it, due to a C-to-T transition at the second base of the second codon of petBIV, was obtained. Biochemical analyses, including electron paramagnetic spectroscopy, with this mutant revealed that the properties of the cytochrome b6c1 complex were similar to those of the cytochrome bc1 complex. In particular, it was highly sensitive to inhibitors of the cytochrome bc1 complex, including antimycin A, and the redox properties of its b- and c-type heme prosthetic groups were unchanged. However, the optical absorption spectrum of its cytochrome bL heme was modified in a way reminiscent of that of a cytochrome b6f complex. Based on the work described here and that with Rhodobacter sphaeroides (R. Kuras, M. Guergova-Kuras, and A. R. Crofts, Biochemistry 37:16280-16288, 1998), it appears that neither the inhibitor resistance nor the redox potential differences observed between the bacterial (or mitochondrial) cytochrome bc1 complexes and the chloroplast cytochrome b6f complexes are direct consequences of splitting cytochrome b into two separate polypeptides. The overall findings also illustrate the possible evolutionary relationships among various cytochrome bc oxidoreductases. PMID- 10464209 TI - Regulation of mga transcription in the group A streptococcus: specific binding of mga within its own promoter and evidence for a negative regulator. AB - Transcription of mga, encoding the multiple virulence gene regulator of the group A streptococcus, is positively autoregulated. This regulation requires a DNA region (Pmga) that contains both a promoter proximal to mga (P2) and a promoter located further upstream (P1). To determine if Mga has a direct role in this process, its ability to bind to specific sequences within Pmga was tested. A purified fusion of Mga to the C-terminal end of maltose-binding protein (MBP Mga), encoded by malE-mga, was shown previously to bind to the promoter regions of Mga-regulated genes, including scpA and emm. We report here that MBP-Mga can function in vivo to regulate emm and mga. Electrophoretic mobility shift assays and DNase I footprinting were used to demonstrate specific binding of MBP-Mga to two ca. 59-bp binding sites in Pmga centered around bases -108 and -180 from the major P2 start of transcription. Mga binding sites from Pemm and PscpA were shown to compete for binding at the two Pmga sites, suggesting that the same domain of Mga interacts at all of these promoter targets. Deletion of the distal Pmga binding site (site I) in vivo resulted in loss of Mga-dependent transcription from the P2 start. However, the same lesion resulted in an increase in P1 transcription that was independent of Mga. This suggests the existence of a repressor of mga transcription with a binding site overlapping those of Mga. PMID- 10464210 TI - Role of the sporulation protein BofA in regulating activation of the Bacillus subtilis developmental transcription factor sigmaK. AB - During sporulation, the Bacillus subtilis transcription factor sigmaK is activated by regulated proteolytic processing. I have used a system that facilitates the analysis of the contributions of a modified form of the processing enzyme, SpoIVFB-GFP, and the regulatory proteins BofA and SpoIVFA to the conversion of pro-sigmaK to sigmaK. The results show that in the presence of BofA, SpoIVFA levels increase by greater than 20-fold, SpoIVFA is substantially stabilized, and pro-sigmaK processing is inhibited. In addition, enhanced accumulation of the SpoIVFA protein in the absence of BofA (achieved through the use of an ftsH null mutation) substantially inhibits pro-sigmaK processing. These results suggest that during growth, increased accumulation of the SpoIVFA protein inhibits the activity of SpoIVFB-GFP and regulates the activation of sigmaK. PMID- 10464211 TI - Regulation of cell component production by growth rate in the group B Streptococcus. AB - Group B Streptococcus (GBS) is the leading cause of bacterial sepsis and meningitis among neonates. While the capsular polysaccharide (CPS) is an important virulence factor of GBS, other cell surface components, such as C proteins, may also play a role in GBS disease. CPS production by GBS type III strain M781 was greater when cells were held at a fast (1.4-h mass-doubling time [td]) than at a slow (11-h td) rate of growth. To further investigate growth rate regulation of CPS production and to investigate production of other cell components, different serotypes and strains of GBS were grown in continuous culture in a semidefined and a complex medium. Samples were obtained after at least five generations at the selected growth rate. Cells and cell-free supernatants were processed immediately, and results from all assays were normalized for cell dry weight. All serotypes (Ia, Ib, and III) and strains (one or two strains per serotype) tested produced at least 3.6-fold more CPS at a td of 1. 4 h than at a td of 11 h. Production of beta C protein by GBS type Ia strain A909 and type Ib strain H36B was also shown to increase at least 5.5-fold with increased growth rate (production at a td of 1. 4 h versus 11 h). The production of alpha C protein by the same strains did not significantly change with increased growth rate. The effect of growth rate on other cell components was also investigated. Production of group B antigen did not change with growth rate, while alkaline phosphatase decreased with increased growth rate. Both CAMP factor and beta-hemolysin production increased fourfold with increased growth rate. Growth rate regulation is specific for select cell components in GBS, including beta C protein, alkaline phosphatase, beta-hemolysin, and CPS production. PMID- 10464212 TI - Comparison of the D-glutamate-adding enzymes from selected gram-positive and gram negative bacteria. AB - The biochemical properties of the D-glutamate-adding enzymes (MurD) from Escherichia coli, Haemophilus influenzae, Enterococcus faecalis, and Staphylococcus aureus were investigated to detect any differences in the activity of this enzyme between gram-positive and gram-negative bacteria. The genes (murD) that encode these enzymes were cloned into pMAL-c2 fusion vector and overexpressed as maltose-binding protein-MurD fusion proteins. Each fusion protein was purified to homogeneity by affinity to amylose resin. Proteolytic treatments of the fusion proteins with factor Xa regenerated the individual MurD proteins. It was found that these fusion proteins retain D-glutamate-adding activity and have Km and Vmax values similar to those of the regenerated MurDs, except for the H. influenzae enzyme. Substrate inhibition by UDP-N-acetylmuramyl L-alanine, the acceptor substrate, was observed at concentrations greater than 15 and 30 microM for E. coli and H. influenzae MurD, respectively. Such substrate inhibition was not observed with the E. faecalis and S. aureus enzymes, up to a substrate concentration of 1 to 2 mM. In addition, the two MurDs of gram-negative origin were shown to require monocations such as NH4+ and/or K+, but not Na+, for optimal activity, while anions such as Cl- and SO4(2-) had no effect on the enzyme activities. The activities of the two MurDs of gram-positive origin, on the other hand, were not affected by any of the ions tested. All four enzymes required Mg2+ for the ligase activity and exhibited optimal activities around pH 8. These differences observed between the gram-positive and gram-negative MurDs indicated that the two gram-negative bacteria may apply a more stringent regulation of cell wall biosynthesis at the early stage of peptidoglycan biosynthesis pathway than do the two gram-positive bacteria. Therefore, the MurD catalyzed reaction may constitute a fine-tuning step necessary for the gram negative bacteria to optimally maintain its relatively thin yet essential cell wall structure during all stages of growth. PMID- 10464213 TI - Roles of RuvC and RecG in phage lambda red-mediated recombination. AB - The recombination properties of Escherichia coli strains expressing the red genes of bacteriophage lambda and lacking recBCD function either by mutation or by expression of lambda gam were examined. The substrates for recombination were nonreplicating lambda chromosomes, introduced by infection; Red-mediated recombination was initiated by a double-strand break created by the action of a restriction endonuclease in the infected cell. In one type of experiment, two phages marked with restriction site polymorphisms were crossed. Efficient formation of recombinant DNA molecules was observed in ruvC+ recG+, ruvC recG+, ruvC+ recG, and ruvC recG hosts. In a second type of experiment, a 1-kb nonhomology was inserted between the double-strand break and the donor chromosome's restriction site marker. In this case, recombinant formation was found to be partially dependent upon ruvC function, especially in a recG mutant background. In a third type of experiment, the recombining partners were the host cell chromosome and a 4-kb linear DNA fragment containing the cat gene, with flanking lac sequences, released from the infecting phage chromosome by restriction enzyme cleavage in the cell; the formation of chloramphenicol resistant bacterial progeny was measured. Dependence on RuvC varied considerably among the three types of cross. However, in all cases, the frequency of Red mediated recombination was higher in recG than in recG+. These observations favor models in which RecG tends to push invading 3'-ended strands back out of recombination intermediates. PMID- 10464214 TI - Two different types of dehalogenases, LinA and LinB, involved in gamma hexachlorocyclohexane degradation in Sphingomonas paucimobilis UT26 are localized in the periplasmic space without molecular processing. AB - gamma-Hexachlorocyclohexane (gamma-HCH) is one of several highly chlorinated insecticides that cause serious environmental problems. The cellular proteins of a gamma-HCH-degrading bacterium, Sphingomonas paucimobilis UT26, were fractionated into periplasmic, cytosolic, and membrane fractions after osmotic shock. Most of two different types of dehalogenase, LinA (gamma hexachlorocyclohexane dehydrochlorinase) and LinB (1,3,4,6-tetrachloro-1,4 cyclohexadiene halidohydrolase), that are involved in the early steps of gamma HCH degradation in UT26 was detected in the periplasmic fraction and had not undertaken molecular processing. Furthermore, immunoelectron microscopy clearly showed that LinA and LinB are periplasmic proteins. LinA and LinB both lack a typical signal sequence for export, so they may be secreted into the periplasmic space via a hitherto unknown mechanism. PMID- 10464215 TI - The frequency of conjugative transposition of Tn916 is not determined by the frequency of excision. AB - Excision and formation of a covalently closed circular transposon molecule are required for conjugative transposition of Tn916 but are not the only factors that limit the frequency of conjugative transposition from one host to another. We found that in gram-positive bacteria, an increase in the frequency of excision and circularization of Tn916 caused by expression of integrase (Int) and excisionase (Xis) from a xylose-inducible promoter does not lead to an increase in the frequency of conjugative transposition. We also found that the concentration of Int and Xis in the recipient cell does not limit the frequency of conjugative transposition and that increased excision does not result in increased expression of transfer functions required to mobilize a plasmid containing the Tn916 origin of transfer. We conclude that in gram-positive hosts in which the Tn916 functions Int and Xis are overexpressed, the frequency of conjugative transposition is limited by the availability of transfer functions. PMID- 10464216 TI - New sporulation loci in Streptomyces coelicolor A3(2). AB - Sporulation mutants of Streptomyces coelicolor appear white because they are defective in the synthesis of the grey polyketide spore pigment, and such white (whi) mutants had been used to define eight sporulation loci, whiA, whiB, whiD, whiE, whiG, whiH, whiI, and whiJ (K. F. Chater, J. Gen. Microbiol. 72:9-28, 1972; N. J. Ryding, Ph.D. thesis, University of East Anglia, 1995). In an attempt to identify new whi loci, we mutagenized S. coelicolor M145 spores with nitrosoguanidine and identified 770 mutants with colonies ranging from white to medium grey. After excluding unstable strains, we examined the isolates by phase contrast microscopy and chose 115 whi mutants with clear morphological phenotypes for further study. To exclude mutants representing cloned whi genes, self transmissible SCP2*-derived plasmids carrying whiA, whiB, whiG, whiH, or whiJ (but not whiD, whiE, or whiI) were introduced into each mutant by conjugation, and strains in which the wild-type phenotype was restored either partially or completely by any of these plasmids were excluded from further analysis. In an attempt to complement some of the remaining 31 whi mutants, an SCP2* library of wild-type S. coelicolor chromosomal DNA was introduced into 19 of the mutants by conjugation. Clones restoring the wild-type phenotype to 12 of the 19 strains were isolated and found to represent five distinct loci, designated whiK, whiL, whiM, whiN, and whiO. Each of the five loci was located on the ordered cosmid library: whiL, whiM, whiN, and whiO occupied positions distinct from previously cloned whi genes; whiK was located on the same cosmid overlap as whiD, but the two loci were shown by complementation to be distinct. The phenotypes resulting from mutations at each of these new loci are described. PMID- 10464217 TI - Amino acid-mediated induction of the basic amino acid-specific outer membrane porin OprD from Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa can utilize arginine and other amino acids as both carbon and nitrogen sources. Earlier studies have shown that the specific porin OprD facilitates the diffusion of basic amino acids as well as the structurally analogous beta-lactam antibiotic imipenem. The studies reported here showed that the expression of OprD was strongly induced when arginine, histidine, glutamate, or alanine served as the sole source of carbon. The addition of succinate exerted a negative effect on induction of oprD, likely due to catabolite repression. The arginine-mediated induction was dependent on the regulatory protein ArgR, and binding of purified ArgR to its operator upstream of the oprD gene was demonstrated by gel mobility shift and DNase assays. The expression of OprD induced by glutamate as the carbon source, however, was independent of ArgR, indicating the presence of more than a single activation mechanism. In addition, it was observed that the levels of OprD responded strongly to glutamate and alanine as the sole sources of nitrogen. Thus, that the expression of oprD is linked to both carbon and nitrogen metabolism of Pseudomonas aeruginosa. PMID- 10464218 TI - Catabolism of branched-chain alpha-keto acids in Enterococcus faecalis: the bkd gene cluster, enzymes, and metabolic route. AB - Genes encoding a branched-chain alpha-keto acid dehydrogenase from Enterococcus faecalis 10C1, E1alpha (bkdA), E1beta (bkdB), E2 (bkdC), and E3 (bkdD), were found to reside in the gene cluster ptb-buk-bkdDABC. The predicted products of ptb and buk exhibited significant homology to the phosphotransbutyrylase and butyrate kinase, respectively, from Clostridium acetobutylicum. Activity and redox properties of the purified recombinant enzyme encoded by bkdD indicate that E. faecalis has a lipoamide dehydrogenase that is distinct from the lipoamide dehydrogenase associated with the pyruvate dehydrogenase complex. Specific activity of the ptb gene product expressed in Escherichia coli was highest with the substrates valeryl-coenzyme A (CoA), isovaleryl-CoA, and isobutyryl-CoA. In cultures, a stoichiometric conversion of alpha-ketoisocaproate to isovalerate was observed, with a concomitant increase in biomass. We propose that alpha ketoisocaproate is converted via the BKDH complex to isovaleryl-CoA and subsequently converted into isovalerate via the combined actions of the ptb and buk gene products with the concomitant phosphorylation of ADP. In contrast, an E. faecalis bkd mutant constructed by disruption of the bkdA gene did not benefit from having alpha-ketoisocaproate in the growth medium, and conversion to isovalerate was less than 2% of the wild-type conversion. It is concluded that the bkd gene cluster encodes the enzymes that constitute a catabolic pathway for branched-chain alpha-keto acids that was previously unidentified in E. faecalis. PMID- 10464219 TI - Mutations affecting motifs of unknown function in the central domain of nitrogen regulatory protein C. AB - The positive control function of the bacterial enhancer-binding protein NtrC resides in its central domain, which is highly conserved among activators of sigma54 holoenzyme. Previous studies of a small set of mutant forms specifically defective in transcriptional activation, called NtrC repressor [NtrC(Rep)] proteins, had enabled us to locate various functional determinants in the central domain. In this more comprehensive survey, the DNA encoding a major portion of the central domain was randomly mutagenized and mutated ntrC genes were introduced into the cell via multicopy expression plasmids. DNA sequencing of 95 isolates identified by a preliminary phenotypic screen revealed that the lesions in them caused 55 distinct single amino acid substitutions at 44 different positions. Assays of glnA transcription in vivo and in vitro yielded two conclusions. First, of the 41 mutant proteins that could be purified, 17 (1 known, 16 new) showed no detectable activity in either assay, thus qualifying them as true NtrC(Rep) proteins. These contained residue changes in six of the seven highly conserved regions in the central domain, including two never studied before. Second, some mutant proteins were inactive in vivo but were either marginally or fully active in vitro. Their surprising lack of activity in vivo may be accounted for by high levels of expression, which apparently decreased activation by these mutant proteins but not by wild-type NtrC (NtrCWT). Of particular interest were a subset of these proteins that exhibited greater transcriptional activation than NtrCWT at low concentrations. Their elevated activation capacities remain to be explained. PMID- 10464220 TI - ZmaR, a novel and widespread antibiotic resistance determinant that acetylates zwittermicin A. AB - ZmaR is a resistance determinant of unusual abundance in the environment and confers on gram-positive and gram-negative bacteria resistance to zwittermicin A, a novel broad-spectrum antibiotic produced by species of Bacillus. The ZmaR protein has no sequence similarity to proteins of known function; thus, the purpose of the present study was to determine the function of ZmaR in vitro. Cell extracts of E. coli containing zmaR inactivated zwittermicin A by covalent modification. Chemical analysis of inactivated zwittermicin A by 1H NMR, 13C NMR, and high- and low-resolution mass spectrometry demonstrated that the inactivated zwittermicin A was acetylated. Purified ZmaR protein inactivated zwittermicin A, and biochemical assays for acetyltransferase activity with [14C]acetyl coenzyme A demonstrated that ZmaR catalyzes the acetylation of zwittermicin A with acetyl coenzyme A as a donor group, suggesting that ZmaR may constitute a new class of acetyltransferases. Our results allow us to assign a biochemical function to a resistance protein that has no sequence similarity to proteins of known function, contributing fundamental knowledge to the fields of antibiotic resistance and protein function. PMID- 10464221 TI - Molecular analysis of the gene encoding a novel transglycosylative enzyme from Alteromonas sp. strain O-7 and its physiological role in the chitinolytic system. AB - We purified from the culture supernatant of Alteromonas sp. strain O-7 and characterized a transglycosylating enzyme which synthesized beta-(1-->6) (GlcNAc)2, 2-acetamido-6-O-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-2- deoxyglucopyranose from beta-(1-->4)-(GlcNAc)2. The gene encoding a novel transglycosylating enzyme was cloned into Escherichia coli, and its nucleotide sequence was determined. The molecular mass of the deduced amino acid sequence of the mature protein was determined to be 99,560 Da which corresponds very closely with the molecular mass of the cloned enzyme determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The molecular mass of the cloned enzyme was much larger than that of enzyme (70 kDa) purified from the supernatant of this strain. These results suggest that the native enzyme was the result of partial proteolysis occurring in the N-terminal region. The enzyme showed significant sequence homology with several bacterial beta-N-acetylhexosaminidases which belong to family 20 glycosyl hydrolases. However, this novel enzyme differs from all reported beta-N-acetylhexosaminidases in its substrate specificity. To clarify the role of the enzyme in the chitinolytic system of the strain, the effect of beta-(1-->6)-(GlcNAc)2 on the induction of chitinase was investigated. beta-(1-->6)-(GlcNAc)2 induced a level of production of chitinase similar to that induced by the medium containing chitin. On the other hand, GlcNAc, (GlcNAc)2, and (GlcNAc)3 conversely repressed the production of chitinase to below the basal level of chitinase activity produced constitutively in medium without a carbon source. PMID- 10464222 TI - Identification of the Omega4499 regulatory region controlling developmental expression of a Myxococcus xanthus cytochrome P-450 system. AB - Omega4499 is the site of a Tn5 lac insertion in the Myxococcus xanthus chromosome that fuses lacZ expression to a developmentally regulated promoter. Cell-cell interactions that occur during development, including C signaling, are required for normal expression of Tn5 lac Omega4499. The DNA upstream of the Omega4499 insertion has been cloned, and the promoter has been localized. Analysis of the DNA sequence downstream of the promoter revealed one complete open reading frame and a second partial open reading frame that is interrupted by Tn5 lac Omega4499. The predicted products of these open reading frames are highly similar to reductase and oxidase components of bacterial cytochrome P-450 systems, which allow catabolism or anabolism of unusual compounds. However, the function of the gene products of the Omega4499 locus remains unclear because M. xanthus containing Tn5 lac Omega4499 exhibits no apparent defect in growth, developmental aggregation, fruiting body formation, or sporulation. Deletion analysis of the Omega4499 regulatory region showed that multiple DNA elements spanning more than 500 bp upstream of the transcriptional start site contribute to developmental promoter activity. At least two DNA elements, one downstream of -49 bp and one between -49 and -218 bp, boosted activity of the promoter in response to intercellular C signaling. Three sequences in the Omega4499 promoter region, centered at -55, -33, and -1 bp, nearly match a 7-bp sequence found in other C signal-dependent promoters. We propose that these sequences, matching the consensus sequence 5'-CAYYCCY-3', be called C box sequences, and we speculate that these sequences are cis-acting regulatory elements important for the expression of M. xanthus genes that depend upon intercellular C signaling during development. PMID- 10464223 TI - Regulation of synthesis of the Bacillus subtilis transition-phase, spore associated antibacterial protein TasA. AB - Previously, we identified a novel component of Bacillus subtilis spores, called TasA, which possesses antibacterial activity. TasA is made early in spore formation, as cells enter stationary phase, and is secreted into the medium as well as deposited into the spore. Here, we show that tasA expression can occur as cells enter stationary phase even under sporulation-repressing conditions, indicating that TasA is a transition-phase protein. tasA and two upstream genes, yqxM and sipW, likely form an operon, transcription of which is under positive control by the transition-phase regulatory genes spo0A and spo0H and negative control by the transition phase regulatory gene abrB. These results are consistent with the suggestion that yqxM, sipW, and tasA constitute a transition phase operon that could play a protective role in a variety of cellular responses to stress during late-exponential-phase and early-stationary-phase growth in B. subtilis. PMID- 10464224 TI - A unique alpha-1,3 mannosyltransferase of the pathogenic fungus Cryptococcus neoformans. AB - The major virulence factor of the pathogenic fungus Cryptococcus neoformans is an extensive polysaccharide capsule which surrounds the cell. Almost 90% of the capsule is composed of a partially acetylated linear alpha-1,3-linked mannan substituted with D-xylose and D-glucuronic acid. A novel mannosyltransferase with specificity appropriate for a role in the synthesis of this glucuronoxylomannan is active in cryptococcal membranes. This membrane-associated activity transfers mannose in vitro from GDP-mannose to an alpha-1, 3-dimannoside acceptor, forming a second alpha-1,3 linkage. Product formation by the transferase is dependent on protein, time, temperature, divalent cations, and each substrate. It is not affected by amphomycin or tunicamycin but is inhibited by GDP and mannose-1 phosphate. The described activity is not detectable in the model yeast Saccharomyces cerevisiae, consistent with the absence of a similar polysaccharide structure in that organism. A second mannosyltransferase from C. neoformans membranes adds mannose in alpha-1,2 linkage to the same dimannoside acceptor. The two activities differ in pH optimum and cation preference. While the alpha-1,2 transferase does not have specificity appropriate for a role in glucuronoxylomannan synthesis, it may participate in production of mannoprotein components of the capsule. This study suggests two new targets for antifungal drug discovery. PMID- 10464225 TI - Characterization of Pseudomonas aeruginosa enoyl-acyl carrier protein reductase (FabI): a target for the antimicrobial triclosan and its role in acylated homoserine lactone synthesis. AB - The Pseudomonas aeruginosa fabI structural gene, encoding enoyl-acyl carrier protein (ACP) reductase, was cloned and sequenced. Nucleotide sequence analysis revealed that fabI is probably the last gene in a transcriptional unit that includes a gene encoding an ATP-binding protein of an ABC transporter of unknown function. The FabI protein was similar in size and primary sequence to other bacterial enoyl-ACP reductases, and it contained signature motifs for the FAD dependent pyridine nucleotide reductase and glucose/ribitol dehydrogenase families, respectively. The chromosomal fabI gene was disrupted, and the resulting mutant was viable but possessed only 62% of the total enoyl-ACP reductase activity found in wild-type cell extracts. The fabI-encoded enoyl-ACP reductase activity was NADH dependent and inhibited by triclosan; the residual activity in the fabI mutant was also NADH dependent but not inhibited by triclosan. An polyhistidine-tagged FabI protein was purified and characterized. Purified FabI (i) could use NADH but not NADPH as a cofactor; (ii) used both crotonyl-coenzyme A and crotonyl-ACP as substrates, although it was sixfold more active with crotonyl-ACP; and (iii) was efficiently inhibited by low concentrations of triclosan. A FabI Gly95-to-Val active-site amino acid substitution was generated by site-directed mutagenesis, and the mutant protein was purified. The mutant FabI protein retained normal enoyl-ACP reductase activity but was highly triclosan resistant. When coupled to FabI, purified P. aeruginosa N-butyryl-L-homoserine lactone (C4-HSL) synthase, RhlI, could synthesize C4-HSL from crotonyl-ACP and S-adenosylmethionine. This reaction was NADH dependent and inhibited by triclosan. The levels of C4-HSL and N-(3-oxo) dodecanoyl-L-homoserine lactones were reduced 50% in a fabI mutant, corroborating the role of FabI in acylated homoserine lactone synthesis in vivo. PMID- 10464227 TI - First evidence for the presence of a hydrogenase in the sulfur-reducing bacterium Desulfuromonas acetoxidans. AB - Hydrogenases, which are ubiquitous in sulfate-reducing bacteria, were previously thought to be absent from Desulfuromonas acetoxidans. For the first time, a hydrogenase from the strict anaerobic sulfur-respiring bacterium D. acetoxidans, grown on ethanol-malate, was detected and enriched. To assay the role of the hydrogenase in the energetic metabolism of D. acetoxidans, we examined the reactivity of the enzyme with polyheme cytochromes from the same bacterium. PMID- 10464226 TI - Characterization of a Pseudomonas aeruginosa fatty acid biosynthetic gene cluster: purification of acyl carrier protein (ACP) and malonyl-coenzyme A:ACP transacylase (FabD). AB - A DNA fragment containing the Pseudomonas aeruginosa fabD (encoding malonyl coenzyme A [CoA]:acyl carrier protein [ACP] transacylase), fabG (encoding beta ketoacyl-ACP reductase), acpP (encoding ACP), and fabF (encoding beta-ketoacyl ACP synthase II) genes was cloned and sequenced. This fab gene cluster is delimited by the plsX (encoding a poorly understood enzyme of phospholipid metabolism) and pabC (encoding 4-amino-4-deoxychorismate lyase) genes; the fabF and pabC genes seem to be translationally coupled. The fabH gene (encoding beta ketoacyl-ACP synthase III), which in most gram-negative bacteria is located between plsX and fabD, is absent from this gene cluster. A chromosomal temperature-sensitive fabD mutant was obtained by site-directed mutagenesis that resulted in a W258Q change. A chromosomal fabF insertion mutant was generated, and the resulting mutant strain contained substantially reduced levels of cis vaccenic acid. Multiple attempts aimed at disruption of the chromosomal fabG gene were unsuccessful. We purified FabD as a hexahistidine fusion protein (H6-FabD) and ACP in its native form via an ACP-intein-chitin binding domain fusion protein, using a novel expression and purification scheme that should be applicable to ACP from other bacteria. Matrix-assisted laser desorption ionization spectroscopy, native polyacrylamide electrophoresis, and amino terminal sequencing revealed that (i) most of the purified ACP was properly modified with its 4'-phosphopantetheine functional group, (ii) it was not acylated, and (iii) the amino-terminal methionine was removed. In an in vitro system, purified ACP functioned as acyl acceptor and H(6)-FabD exhibited malonyl CoA:ACP transacylase activity. PMID- 10464228 TI - The Escherichia coli NadR regulator is endowed with nicotinamide mononucleotide adenylyltransferase activity. AB - The first identification and characterization of a catalytic activity associated with NadR protein is reported. A computer-aided search for sequence similarity revealed the presence in NadR of a 29-residue region highly conserved among known nicotinamide mononucleotide adenylyltransferases. The Escherichia coli nadR gene was cloned into a T7-based vector and overexpressed. In addition to functionally specific DNA binding properties, the homogeneous recombinant protein catalyzes NAD synthesis from nicotinamide mononucleotide and ATP. PMID- 10464229 TI - Genomic plasticity in natural populations of Bordetella pertussis. AB - We determined the genomic organization of 14 clinical strains of Bordetella pertussis isolated over an 18-month period in Alberta, Canada. The maps of these 14 strains, while demonstrating general similarity of gene order, display a number of examples of genomic rearrangements in the form of large chromosomal inversions. PMID- 10464230 TI - Molecular characterization of the PhoP-PhoQ two-component system in Escherichia coli K-12: identification of extracellular Mg2+-responsive promoters. AB - We identified Mg2+-responsive promoters of the phoPQ, mgtA, and mgrB genes of Escherichia coli K-12 by S1 nuclease analysis. Expression of these genes was induced by magnesium limitation and depended on PhoP and PhoQ. The transcription start sites were also determined, which allowed us to find a (T/G)GTTTA direct repeat in their corresponding promoter regions. PMID- 10464231 TI - Acetate utilization in Lactococcus lactis deficient in lactate dehydrogenase: a rescue pathway for maintaining redox balance. AB - Acetate was shown to improve glucose fermentation in Lactococcus lactis deficient in lactate dehydrogenase. 13C and 1H nuclear magnetic resonance studies using [2 13C]glucose and [2-(13)C]acetate as substrates demonstrated that acetate was exclusively converted to ethanol. This novel pathway provides an alternative route for NAD+ regeneration in the absence of lactate dehydrogenase. PMID- 10464232 TI - Clustering of the chemoreceptor complex in Escherichia coli is independent of the methyltransferase CheR and the methylesterase CheB. AB - The Escherichia coli chemoreceptors and their associated cytoplasmic proteins, CheA and CheW, cluster predominantly at the cell poles. The nature of the clustering remains a mystery. Recent studies suggest that CheR binding to and/or methylation of the chemoreceptors may play a role in chemoreceptor complex aggregation. In this study, we examined the intracellular distribution of the chemoreceptors by immunoelectron microscopy in strains lacking either the methyltransferase CheR or the methylesterase CheB. The localization data revealed that, in vivo, aggregation of the chemoreceptor complex was independent of either CheR or CheB. PMID- 10464233 TI - A hyperactive NAD(P)H:Rubredoxin oxidoreductase from the hyperthermophilic archaeon Pyrococcus furiosus. AB - NAD(P)H:rubredoxin oxidoreductase (NROR) has been purified from the hyperthermophilic archaeon Pyrococcus furiosus. The enzyme is exceedingly active in catalyzing the NADPH-dependent reduction of rubredoxin, a small (5.3-kDa) iron containing redox protein that had previously been purified from this organism. The apparent Vmax at 80 degrees C is 20,000 micromol/min/mg, which corresponds to a kcat/Km value of 300,000 mM(-1) s(-1). The apparent Km values measured at 80 degrees C and pH 8.0 for rubredoxin, NADPH, and NADH were 50, 5, and 34 microM, respectively. The enzyme did not reduce P. furiosus ferredoxin. NROR is a monomer with a molecular mass of 45 kDa and contains one flavin adenine dinucleotide molecule per mole but lacks metals and inorganic sulfide. The possible physiological role of this hyperactive enzyme is discussed. PMID- 10464234 TI - Identification of a conserved N-terminal sequence involved in transmembrane signal transduction in EnvZ. AB - To determine whether N-terminal sequences are involved in the transmembrane signaling mechanism of EnvZ, the nucleotide sequences of envZ genes from several enteric bacteria were determined. Comparative analysis revealed that the amino acid sequence between Pro41 and Glu53 was highly conserved. To further analyze the role of the conserved sequence, envZ of Escherichia coli was subjected to random PCR mutagenesis and mutant alleles that produced a high-osmolarity phenotype, in which ompF was repressed, were isolated. The mutations identified clustered within, as well as adjacent to, the Pro41-to-Glu53 sequence. These findings suggest that the conserved Pro41-to-Glu53 sequence is involved in the signal transduction mechanism of EnvZ. PMID- 10464236 TI - Biologically active oxidized phospholipids. PMID- 10464235 TI - A new class of cobalamin transport mutants (btuF) provides genetic evidence for a periplasmic binding protein in Salmonella typhimurium. AB - No periplasmic binding protein has been demonstrated for the ATP-binding cassette (ABC)-type cobalamin transporter BtuCD. New mutations (btuF) are described that affect inner-membrane transport. The BtuF protein has a signal sequence and resembles the periplasmic binding proteins of several other ABC transporters. PMID- 10464237 TI - Phospholipid membranes form specific nonbilayer molecular arrangements that are antigenic. AB - Hexagonal phase (H(II))-preferring lipids such as phosphatidate, cardiolipin, and phosphatidylserine form nonbilayer molecular arrangements in lipid bilayers. While their presence in biological membranes has not been established, in vitro studies suggest that alterations in membrane properties modify their function. In this study, antiphospholipid monoclonal antibodies were developed against nonbilayer structures. One of the monoclonal antibodies identifies nonplanar surfaces in liposomes and in membranes of cultured cells. These results are the first evidence that natural membranes maintain a fragile balance between bilayer and nonbilayer lipid arrangements. Therefore, these antibodies can be used to evaluate the role of H(II)-preferring lipids in the modulation of membrane activities. Our studies demonstrated that nonplanar surfaces are highly immunogenic. Although these structures are normally transient, their formation can be stabilized by temperature variations, drugs, antibiotics, apolar peptides, and divalent cations. Our studies demonstrated that abnormal exposure of nonbilayer arrangements may induce autoimmune responses as found in the antiphospholipid syndrome. PMID- 10464238 TI - Association of frabin with the actin cytoskeleton is essential for microspike formation through activation of Cdc42 small G protein. AB - We have recently isolated a novel actin filament-binding protein, named frabin. Frabin has one actin filament-binding domain (ABD), one Dbl homology domain (DHD), first pleckstrin homology domains (PHD) adjacent to DHD, one cysteine rich domain (CRD), and second PHD from the N terminus to the C terminus in this order. Full-length frabin induces microspike formation and c-Jun N-terminal kinase (JNK) activation. We found here that the fragment of frabin containing DHD and first PHD stimulated guanine nucleotide exchange of Cdc42Hs small G protein, but not that of RhoA or Rac1 small G protein. However, this fragment of frabin did not induce microspike formation, and ABD was additionally necessary for microspike formation. Frabin having ABD was associated with the actin cytoskeleton, whereas frabin lacking ABD was diffusely distributed in the cytoplasm. In contrast, ABD was not necessary for JNK activation but CRD and second PHD were additionally necessary for this activation. These results indicate that the association of frabin with the actin cytoskeleton is essential for microspike formation but not for JNK activation and that different domains of frabin are involved in microspike formation and JNK activation through Cdc42 activation. PMID- 10464239 TI - The envelope anion channel involved in chloroplast protein import is associated with Tic110. AB - An anion channel of the chloroplast envelope was previously shown to be involved in protein import. Some gating characteristics of the channel are presented. The pore size of the channel is estimated to be around 6.5 A. Antibodies raised to Tic110 completely inactivate the protein import-related channel. These observations suggest that the channel is associated with the Tic machinery and can function as the protein conducting channel of the inner envelope membrane. PMID- 10464240 TI - cDNA cloning of a novel androgen receptor subtype. AB - There has been general acceptance that only one type of androgen receptor (AR) exists in an individual. This contrasts with other members of the nuclear receptor superfamily where multiple forms have been reported (e.g. estrogen receptor alpha/beta, thyroid hormone receptor alpha/beta, etc.). We have previously identified 11-ketotestosterone (a potent androgen in teleosts) as the spermatogenesis-inducing hormone of the Japanese eel and have cloned its receptor (eAR1) cDNA from eel testis. Here we report on the cloning of a cDNA encoding a second type of AR (eAR2) from the eel testis and the functional characterization of the encoded protein. This cDNA contains a complete open reading frame encoding 797 amino acid residues. The amino acid sequence of eAR2 shows high homology with other ARs, including eAR1, in the DNA-binding (98-88%) and ligand-binding (59 85%) domains, whereas the other domains show low homology (<35%). In transient transfection assays of mammalian cells, the eAR2 protein displayed androgen dependent activation of transcription from the androgen-responsive murine mammary tumor virus promoter. Tissue distribution of its mRNA was different from that of eAR1. We conclude that eAR2 is a novel AR in the eel, which we suggest should be named eel ARbeta to distinguish it from eAR1 (eARalpha). PMID- 10464241 TI - Insulin regulation of protein traffic in rat adipose cells. AB - Rat adipocytes were biotinylated with cell-impermeable reagents, sulfo-N hydroxysuccinimide-biotin and sulfo-N-hydroxysuccinimide-S-S-biotin in the absence and presence of insulin. Biotinylated and nonbiotinylated populations of the insulin-like growth factor-II/mannose 6-phosphate receptor, the transferrin receptor, and insulin-responsive aminopeptidase were separated by adsorption to streptavidin-agarose to determine the percentage of the biotinylated protein molecules versus their total amount in different subcellular compartments. Results indicate that adipose cells possess at least two distinct cell surface recycling pathways for insulin-like growth factor-II/mannose 6-phosphate receptor (MPR) and transferrin receptor (TfR): one which is mediated by glucose transporter isoform 4(Glut4)-vesicles and another that bypasses this compartment. Under basal conditions, the first pathway is not active, and cell surface recycling of TfR and, to a lesser extent, MPR proceeds via the second pathway. Insulin dramatically stimulates recycling through the first pathway and has little effect on the second. Within the Glut4-containing compartment, insulin has profoundly different effects on intracellular trafficking of insulin-responsive aminopeptidase on one hand and MPR and TfR on the other. After insulin administration, insulin-responsive aminopeptidase is redistributed from Glut4 containing vesicles to the plasma membrane and stays there for at least 30 min with minimal detectable internalization and recycling, whereas MPR and TfR rapidly shuttle between Glut4 vesicles and the plasma membrane in such a way that after 30 min of insulin treatment, virtually every receptor molecule in this compartment completes at least one trafficking cycle to the cell surface. Thus, different recycling proteins, which compose Glut4-containing vesicles, are internalized into this compartment at their own distinctive rates. PMID- 10464242 TI - L-arginine binding to nitric-oxide synthase. The role of H-bonds to the nonreactive guanidinium nitrogens. AB - Nitric-oxide synthase (NOS) catalyzes the oxidation of L-arginine to nitric oxide and L-citrulline. Because overproduction of nitric oxide causes tissue damage in neurological, inflammatory, and autoimmune disorders, design of NOS inhibitors has received much attention. Most inhibitors described to date include a guanidine-like structural motif and interact with the guanidinium region of the L arginine-binding site. We report here studies with L-arginine analogs having one or both terminal guanidinium nitrogens replaced by functionalities that preserve some, but not all, of the molecular interactions possible for the -NH(2), =NH, or =NH(2)(+) groups of L-arginine. Replacement groups include -NH-alkyl, -alkyl, =O, and =S. Binding of L-canavanine, an analog unable to form hydrogen bonds involving a N(5)-proton, was also examined. From our results and previous work, we infer the orientation of these compounds in the L-arginine-binding site and use IC(50) or K(i) values and optical difference spectra to quantitate their affinity relative to L-arginine. We find that the non-reactive guanidinium nitrogen of L-arginine binds in a pocket that is relatively intolerant of changes in the size or hydrogen bonding properties of the group bound. The individual H bonds involved are, however, weaker than expected (<2 versus 3-6 kcal). These findings elucidate substrate binding forces in the NOS active site and identify an important constraint on NOS inhibitor design. PMID- 10464243 TI - Interaction of nucleotides with Asp(351) and the conserved phosphorylation loop of sarcoplasmic reticulum Ca(2+)-ATPase. AB - The nucleotide binding properties of mutants with alterations to Asp(351) and four of the other residues in the conserved phosphorylation loop, (351)DKTGTLT(357), of sarcoplasmic reticulum Ca(2+)-ATPase were investigated using an assay based on the 2', 3'-O-(2,4,6-trinitrophenyl)-8-azidoadenosine triphosphate (TNP-8N(3)-ATP) photolabeling of Lys(492) and competition with ATP. In selected cases where the competition assay showed extremely high affinity, ATP binding was also measured by a direct filtration assay. At pH 8.5 in the absence of Ca(2+), mutations removing the negative charge of Asp(351) (D351N, D351A, and D351T) produced pumps that bound MgTNP-8N(3)-ATP and MgATP with affinities 20-156 fold higher than wild type (K(D) as low as 0.006 microM), whereas the affinity of mutant D351E was comparable with wild type. Mutations K352R, K352Q, T355A, and T357A lowered the affinity for MgATP and MgTNP-8N(3)-ATP 2-1000- and 1-6-fold, respectively, and mutation L356T completely prevented photolabeling of Lys(492). In the absence of Ca(2+), mutants D351N and D351A exhibited the highest nucleotide affinities in the presence of Mg(2+) and at alkaline pH (E1 state). The affinity of mutant D351A for MgATP was extraordinarily high in the presence of Ca(2+) (K(D) = 0.001 microM), suggesting a transition state like configuration at the active site under these conditions. The mutants with reduced ATP affinity, as well as mutants D351N and D351A, exhibited reduced or zero CrATP-induced Ca(2+) occlusion due to defective CrATP binding. PMID- 10464244 TI - Binding of correolide to K(v)1 family potassium channels. Mapping the domains of high affinity interaction. AB - Correolide, a novel nortriterpene natural product, potently inhibits the voltage gated potassium channel, K(v)1.3, and [(3)H]dihydrocorreolide (diTC) binds with high affinity (K(d) approximately 10 nM) to membranes from Chinese hamster ovary cells that express K(v)1.3 (Felix, J. P., Bugianesi, R. M., Schmalhofer, W. A., Borris, R., Goetz, M. A., Hensens, O. D., Bao, J.-M., Kayser, F. , Parsons, W. H., Rupprecht, K., Garcia, M. L., Kaczorowski, G. J., and Slaughter, R. S. (1999) Biochemistry 38, 4922-4930). Mutagenesis studies were used to localize the diTC binding site and to design a high affinity receptor in the diTC-insensitive channel, K(v)3.2. Transferring the pore from K(v)1.3 to K(v)3.2 produces a chimera that binds peptidyl inhibitors of K(v)1.3 with high affinity, but not diTC. Transfer of the S(5) region of K(v)1.3 to K(v)3.2 reconstitutes diTC binding at 4-fold lower affinity as compared with K(v)1.3, whereas transfer of the entire S(5)-S(6) domain results in a normal K(v)1.3 phenotype. Substitutions in S(5)-S(6) of K(v)1.3 with nonconserved residues from K(v)3.2 has identified two positions in S(5) and one in S(6) that cause significant alterations in diTC binding. High affinity diTC binding can be conferred to K(v)3.2 after substitution of these three residues with the corresponding amino acids found in K(v)1.3. These results suggest that lack of sensitivity of K(v)3.2 to diTC is a consequence of the presence of Phe(382) and Ile(387) in S(5), and Met(458) in S(6). Inspection of K(v)1.1-1.6 channels indicates that they all possess identical S(5) and S(6) domains. As expected, diTC binds with high affinity (K(d) values 7-21 nM) to each of these homotetrameric channels. However, the kinetics of binding are fastest with K(v)1.3 and K(v)1.4, suggesting that conformations associated with C-type inactivation will facilitate entry and exit of diTC at its binding site. Taken together, these findings identify K(v)1 channel regions necessary for high affinity diTC binding, as well as, reveal a channel conformation that markedly influences the rate of binding of this ligand. PMID- 10464245 TI - Integration of Rac-dependent regulation of cyclin D1 transcription through a nuclear factor-kappaB-dependent pathway. AB - The small GTP-binding protein Rac1, a member of the Ras superfamily, plays a fundamental role in cytoskeleton reorganization, cellular transformation, the induction of DNA synthesis, and superoxide production. Cyclin D1 abundance is rate-limiting in normal G(1) phase progression, and the abundance of cyclin D1 is induced by activating mutations of both Ras and Rac1. Nuclear factor-kappaB (NF kappaB) proteins consist of cytoplasmic hetero- or homodimeric Rel-related proteins complexed to a member of the IkappaB family of inhibitor proteins. In the current studies, activating mutants of Rac1 (Rac(Leu-61), Rac(Val-12)) induced cyclin D1 expression and the cyclin D1 promoter in NIH 3T3 cells. Induction of cyclin D1 by Rac1 required both an NF-kappaB and an ATF-2 binding site. Inhibiting NF-kappaB by overexpression of an NF-kappaB trans-dominant inhibitor (nonphosphorylatable IkappaBalpha) reduced cyclin D1 promoter activation by the Rac1 mutants, placing NF-kappaB in a pathway of Rac1 activation of cyclin D1. Specific amino acid mutations in the amino-terminal effector domain of Rac(Leu-61) had comparable effects on NF-kappaB transcriptional activity and activation of the cyclin D1 promoter. The NF-kappaB factors Rel A (p65) and NF kappaB(1) (p50) induced the cyclin D1 promoter, requiring both the NF-kappaB binding site and the ATF-2 site. Stable overexpression of Rac(Leu-61) increased binding of Rel A and NF-kappaB(1) to the cyclin D1 promoter NF-kappaB site. Activation of Rac1 in NIH 3T3 cells induces both NF-kappaB binding and activity and enhances expression of cyclin D1 through an NF-kappaB and ATF-2 site in the proximal promoter, suggesting a critical role for NF-kappaB in cell cycle regulation through cyclin D1 and Rac1. PMID- 10464246 TI - Effect of glucose on endothelin-1-induced calcium transients in cultured bovine retinal pericytes. AB - Published work has shown that endothelin-1-induced contractility of bovine retinal pericytes is reduced after culture in high concentrations of glucose. The purpose of the present study was to establish the profile of endothelin-1-induced calcium transients in pericytes and to identify changes occurring after culture in high concentrations of glucose. Glucose had no effect on basal levels of cytosolic calcium or on endothelin-1-induced calcium release from intracellular stores. However, influx of calcium from the extracellular medium after endothelin 1 stimulation was reduced in pericytes that had been cultured in 25 mM D-glucose. L-type Ca(2+) currents were identified by patch clamping. The L-type Ca(2+) channel agonist, (-)-Bay K8644, caused less influx of calcium from the extracellular medium in pericytes that had been cultured in 25 mM D-glucose than in those cultured with 5 mM D-glucose. However, 3-O-methylglucose, a nonmetabolizable analogue of glucose which can cause glycation, had similar effects to those of high concentrations of glucose. The results suggest that reduced function of the L-type Ca(2+) channel that occurs in bovine retinal pericytes after culture in high concentrations of D-glucose is probably due to glycation of a channel protein. PMID- 10464247 TI - Importance of redox potential for the in vivo function of the cytoplasmic disulfide reductant thioredoxin from Escherichia coli. AB - The thioredoxin superfamily consists of enzymes that catalyze the reduction, formation, and isomerization of disulfide bonds and exert their activity through a redox active disulfide in a Cys-Xaa(1)-Xaa(2)-Cys motif. The individual members of the family differ strongly in their intrinsic redox potentials. However, the role of the different redox potentials for the in vivo function of these enzymes is essentially unknown. To address the question of in vivo importance of redox potential for the most reducing member of the enzyme family, thioredoxin, we have employed a set of active site variants of thioredoxin with increased redox potentials (-270 to -195 mV) for functional studies in the cytoplasm of Escherichia coli. The variants proved to be efficient substrates of thioredoxin reductase, providing a basis for an in vivo characterization of NADPH-dependent reductive processes catalyzed by the thioredoxin variants. The reduction of sulfate and methionine sulfoxide, as well as the isomerization of periplasmic disulfide bonds by DsbC, which all depend on thioredoxin as catalyst in the E. coli cytoplasm, proved to correlate well with the intrinsic redox potentials of the variants in complementation assays. The same correlation could be established in vitro by using the thioredoxin-catalyzed reduction of lipoic acid by NADPH as a model reaction. We propose that the rate of direct reduction of substrates by thioredoxin, which largely depends on the redox potential of thioredoxin, is the most important parameter for the in vivo function of thioredoxin, as recycling of reduced thioredoxin through NADPH and thioredoxin reductase is not rate-limiting for its catalytic cycle. PMID- 10464248 TI - Inhibition of the broad spectrum nonmetallocarbapenamase of class A (NMC-A) beta lactamase from Enterobacter cloacae by monocyclic beta-lactams. AB - beta-Lactamases hydrolyze beta-lactam antibiotics, a reaction that destroys their antibacterial activity. These enzymes, of which four classes are known, are the primary cause of resistance to beta-lactam antibiotics. The class A beta lactamases form the largest group. A novel class A beta-lactamase, named the nonmetallocarbapenamase of class A (NMC-A) beta-lactamase, has been discovered recently that has a broad substrate profile that included carbapenem antibiotics. This is a serious development, since carbapenems have been relatively immune to the action of these resistance enzymes. Inhibitors for this enzyme are sought. We describe herein that a type of monobactam molecule of our design inactivates the NMC-A beta-lactamase rapidly, efficiently, and irreversibly. The mechanism of inactivation was investigated by solving the x-ray structure of the inhibited NMC A enzyme to 1.95 A resolution. The structure shed light on the nature of the fragmentation of the inhibitor on enzyme acylation and indicated that there are two acyl-enzyme species that account for enzyme inhibition. Each of these inhibited enzyme species is trapped in a distinct local energy minimum that does not predispose the inhibitor species for deacylation, accounting for the irreversible mode of enzyme inhibition. Molecular dynamics simulations provided evidence in favor of a dynamic motion for the acyl-enzyme species, which samples a considerable conformational space prior to the entrapment of the two stable acyl-enzyme species in the local energy minima. A discussion of the likelihood of such dynamic motion for turnover of substrates during the normal catalytic processes of the enzyme is presented. PMID- 10464250 TI - Cyclic strain stress-induced mitogen-activated protein kinase (MAPK) phosphatase 1 expression in vascular smooth muscle cells is regulated by Ras/Rac-MAPK pathways. AB - Recently, we demonstrated that mechanical stress results in rapid phosphorylation or activation of platelet-derived growth factor receptors in vascular smooth muscle cells (VSMCs) followed by activation of mitogen-activated protein kinases (MAPKs) and AP-1 transcription factors (Hu, Y., Bock, G., Wick, G., and Xu, Q. (1998) FASEB J. 12, 1135-1142). Herein, we provide evidence that VSMC responses to mechanical stress also include induction of MAPK phosphatase-1 (MKP-1), which may serve as a negative regulator of MAPK signaling pathways. When rat VSMCs cultivated on a flexible membrane were subjected to cyclic strain stress (60 cycles/min, 5-30% elongation), induction of MKP-1 proteins and mRNA was observed in time- and strength-dependent manners. Concomitantly, mechanical forces evoked rapid and transient activation of all three members of MAPKs, i.e. extracellular signal-regulated kinases (ERKs), c-Jun NH(2)-terminal protein kinases (JNKs), or stress-activated protein kinases (SAPKs), and p38 MAPKs. Suramin, a growth factor receptor antagonist, completely abolished ERK activation, significantly blocked MKP-1 expression, but not JNK/SAPK and p38 MAPK activation, in response to mechanical stress. Interestingly, VSMC lines stably expressing dominant negative Ras (Ras N17) or Rac (Rac N17) exhibited a marked decrease in MKP-1 expression; the inhibition of ERK kinases (MEK1/2) by PD 98059 or of p38 MAPKs by SB 202190 resulted in a down-regulation of MKP-1 induction. Furthermore, overexpressing MKP 1 in VSMCs led to the dephosphorylation and inactivation of ERKs, JNKs/SAPKs, and p38 MAPKs and inhibition of DNA synthesis. Taken together, our findings demonstrate that mechanical stress induces MKP-1 expression regulated by two signal pathways, including growth factor receptor-Ras-ERK and Rac-JNK/SAPK or p38 MAPK, and that MKP-1 inhibits VSMC proliferation via MAPK inactivation. These results suggest that MKP-1 plays a crucial role in mechanical stress-stimulated signaling leading to VSMC growth and differentiation. PMID- 10464249 TI - Protein-RNA interactions determine the stability of the renal NaPi-2 cotransporter mRNA and its translation in hypophosphatemic rats. AB - Hypophosphatemia leads to an increase in type II Na(+)-dependent inorganic phosphate cotransporter (NaPi-2) mRNA and protein levels in the kidney and increases renal phosphate reabsorption. Nuclear transcript run-on experiments showed that the effect of a low phosphate diet was post-transcriptional. In an in vitro degradation assay, renal proteins from hypophosphatemic rats stabilized the NaPi-2 transcript 6-fold compared with control rats and this was dependent upon an intact NaPi-2 3'-untranslated region (UTR). To determine an effect of hypophosphatemia upon NaPi-2 protein synthesis, the incorporation of injected [(35)S]methionine into renal proteins was studied in vivo. Hypophosphatemia led to increased [(35)S]methionine incorporation only into NaPi-2 protein. The effect of hypophosphatemia on translation was studied in an in vitro translation assay, where hypophosphatemic renal proteins led to increased translation of NaPi-2 and other transcripts. NaPi-2 RNA interaction with cytosolic proteins was studied by UV cross-linking and Northwestern gels. Hypophosphatemic proteins led to increased binding of renal cytosolic proteins to the 5'-UTR of NaPi-2 mRNA. Therefore, hypophosphatemia increases NaPi-2 gene expression post transcriptionally, which correlates with a more stable transcript mediated by the 3'-UTR, and an increase in NaPi-2 translation involving protein binding to the 5' UTR. These findings show that phosphate regulates gene expression by affecting protein-RNA interactions in vivo. PMID- 10464251 TI - The proteolipid of the A(1)A(0) ATP synthase from Methanococcus jannaschii has six predicted transmembrane helices but only two proton-translocating carboxyl groups. AB - The proteolipid, a hydrophobic ATPase subunit essential for ion translocation, was purified from membranes of Methanococcus jannaschii by chloroform/methanol extraction and gel chromatography and was studied using molecular and biochemical techniques. Its apparent molecular mass as determined in SDS-polyacrylamide gel electrophoresis varied considerably with the conditions applied. The N-terminal sequence analysis made it possible to define the open reading frame and revealed that the gene is a triplication of the gene present in bacteria. In some of the proteolipids, the N-terminal methionine is excised. Consequently, two forms with molecular masses of 21,316 and 21,183 Da were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The molecular and biochemical data gave clear evidence that the mature proteolipid from M. jannaschii is a triplication of the 8-kDa proteolipid present in bacterial F(1)F(0) ATPases and most archaeal A(1)A(0) ATPases. Moreover, the triplicated form lacks a proton-translocating carboxyl group in the first of three pairs of transmembrane helices. This finding puts in question the current view of the evolution of H(+) ATPases and has important mechanistic consequences for the structure and function of H(+) ATPases in general. PMID- 10464252 TI - Functions of the sigma(54) region I in trans and implications for transcription activation. AB - Control of transcription frequently involves the direct interaction of activators with RNA polymerase. In bacteria, the formation of stable open promoter complexes by the sigma(54) RNA polymerase is critically dependent on sigma(54) amino Region I sequences. Their presence correlates with activator dependence, and removal allows the holoenzyme to engage productively with melted DNA independently of the activator. Using purified Region I sequences and holoenzymes containing full length or Region I-deleted sigma(54), we have explored the involvement of Region I in transcription activation. Results show that Region I in trans inhibits a reversible conformational change in the holoenzyme believed to be polymerase isomerization. Evidence is presented indicating that the holoenzyme (and not the promoter DNA per se) is one interacting target used by Region I in preventing polymerase isomerization. Activator overcomes this inhibition in a reaction requiring nucleotide hydrolysis. Region I in trans is able to inhibit activated transcription by the holoenzyme containing full-length sigma(54). Inhibition appeared to be noncompetitive with respect to the activator, suggesting that a direct activator interaction occurs with parts of the holoenzyme outside Region I. Stabilization of isomerized holoenzyme bound to melted DNA by Region I in trans occurs largely independently of the initiating nucleotide, suggesting a role for Region I in maintaining the open complex. PMID- 10464253 TI - S100A12 is expressed exclusively by granulocytes and acts independently from MRP8 and MRP14. AB - Changes in cytosolic calcium concentrations regulate a wide variety of cellular processes, and calcium-binding proteins are the key molecules in signal transduction, differentiation, and cell cycle control. S100A12, a recently described member of the S100 protein family, has been shown to be coexpressed in granulocytes and monocytes together with two other S100 proteins, MRP8 (S100A8) and MRP14 (S100A9), and a functional relationship between these three S100 proteins has been suggested. Using Western blotting, calcium overlays, intracellular flow cytometry, and cytospin preparations, we demonstrate that S100A12 expression in leukocytes is specifically restricted to granulocytes and that S100A12 represents one of the major calcium-binding proteins in these cells. S100A12, MRP8, and MRP14 translocate simultaneously from the cytosol to cytoskeletal and membrane structures in a calcium-dependent manner. However, no evidence for direct protein-protein interactions of S100A12 with either MRP8 or MRP14 or the heterodimer was found by chemical cross-linking, density gradient centrifugation, mass spectrometric measurements, or yeast two hybrid detection. Thus, S100A12 acts individually during calcium-dependent signaling, independent of MRP8, MRP14, and the heterodimer MRP8/MRP14. This granulocyte-specific signal transduction pathway may offer attractive targets for therapeutic intervention with exaggerated granulocyte activity in pathological states. PMID- 10464254 TI - ADP-ribosylation factor 6 (ARF6) defines two insulin-regulated secretory pathways in adipocytes. AB - ADP-ribosylation factor 6 (ARF6) appears to play an essential role in the endocytic/recycling pathway in several cell types. To determine whether ARF6 is involved in insulin-regulated exocytosis, 3T3-L1 adipocytes were infected with recombinant adenovirus expressing wild-type ARF6 or an ARF6 dominant negative mutant (D125N) that encodes a protein with nucleotide specificity modified from guanine to xanthine. Overexpression of these ARF6 proteins affected neither basal nor insulin-regulated glucose uptake in 3T3-L1 adipocytes, nor did it affect the subcellular distribution of Glut1 or Glut4. In contrast, the secretion of adipsin, a serine protease specifically expressed in adipocytes, was increased by the expression of wild-type ARF6 and was inhibited by the expression of D125N. These results indicate a requirement for ARF6 in basal and insulin-regulated adipsin secretion but not in glucose transport. Our results suggest the existence of at least two distinct pathways that undergo insulin-stimulated exocytosis in 3T3-L1 adipocytes, one for adipsin release and one for glucose transporter translocation. PMID- 10464255 TI - The platelet cytoskeleton regulates the affinity of the integrin alpha(IIb)beta(3) for fibrinogen. AB - Agonist-generated inside-out signals enable the platelet integrin alpha(IIb)beta(3) to bind soluble ligands such as fibrinogen. We found that inhibiting actin polymerization in unstimulated platelets with cytochalasin D or latrunculin A mimics the effects of platelet agonists by inducing fibrinogen binding to alpha(IIb)beta(3). By contrast, stabilizing actin filaments with jasplakinolide prevented cytochalasin D-, latrunculin A-, and ADP-induced fibrinogen binding. Cytochalasin D- and latrunculin A-induced fibrinogen was inhibited by ADP scavengers, suggesting that subthreshold concentrations of ADP provided the stimulus for the actin filament turnover required to see cytochalasin D and latrunculin A effects. Gelsolin, which severs actin filaments, is activated by calcium, whereas the actin disassembly factor cofilin is inhibited by serine phosphorylation. Consistent with a role for these factors in regulating alpha(IIb)beta(3) function, cytochalasin D- and latrunculin A-induced fibrinogen binding was inhibited by the intracellular calcium chelators 1,2-bis(2 aminophenoxy)ethane-N,N,N', N'-tetraacetic acid acetoxymethyl ester and EGTA acetoxymethyl ester and the Ser/Thr phosphatase inhibitors okadaic acid and calyculin A. Our results suggest that the actin cytoskeleton in unstimulated platelets constrains alpha(IIb)beta(3) in a low affinity state. We propose that agonist-stimulated increases in platelet cytosolic calcium initiate actin filament turnover. Increased actin filament turnover then relieves cytoskeletal constraints on alpha(IIb)beta(3), allowing it to assume the high affinity conformation required for soluble ligand binding. PMID- 10464257 TI - Heparin dodecasaccharide binding to platelet factor-4 and growth-related protein alpha. Induction of a partially folded state and implications for heparin-induced thrombocytopenia. AB - alpha-Chemokines are known heparin-binding proteins. Here, a heparin dodecasaccharide (H12) was purified and used in NMR studies to investigate binding to growth-related protein-alpha (Gro-alpha) and to platelet factor-4-M2 (PF4-M2), an N-terminal chimera of PF4. Pulsed field gradient NMR was used to derive diffusion coefficients as the protein (monomer):H12 ratio was varied. In the absence of H12, both PF4-M2 and Gro-alpha give diffusion coefficients consistent with the presence of mostly dimers. As the PF4-M2:H12 ratio is increased from 1:6 to 2:1, the diffusion coefficient increases, indicating dissociation to the monomer state. On addition of H12 to either protein, (15)N/(1)H heteronuclear single quantum coherence NMR data demonstrate loss of (1)H resonance dispersion and intensity, particularly at protein:H12 ratios of 2:1 to 4:1, indicating significant perturbation to native structures. For Gro alpha in particular, (1)H resonance dispersion appears random coil-like. At these same ratios, circular dichroism (CD) data show general retention of secondary structure elements with a slight shift to additional helix formation. Random coil NMR resonance dispersion suggests a shift to a less compact, partially folded, and/or more flexible state. Further addition of H12 causes resonance intensity and dispersion to return making NMR spectra appear native-like. At low PF4-M2:H12 ratios, loss of resonance intensity for residues proximal to Arg-20 and Arg-22 in three-dimensional NMR HCCH-TOCSY spectra suggests that the Arg-20-Arg-22 loop either interacts most strongly with H12 and/or that binding at this site is heterogeneous. This domain was previously shown to be crucial to heparin binding. Of particular interest to the biology of PF4-heparin complex formation, heparin induced thrombocytopenia antibody binding occurs at about the same PF4-M2:H12 ratio as does this transition to a partially folded PF4-M2 state, strongly suggesting that heparin-induced thrombocytopenia antibody recognizes a less folded, lower aggregate state of the protein. PMID- 10464256 TI - Insulin activates protein kinases C-zeta and C-lambda by an autophosphorylation dependent mechanism and stimulates their translocation to GLUT4 vesicles and other membrane fractions in rat adipocytes. AB - In rat adipocytes, insulin provoked rapid increases in (a) endogenous immunoprecipitable combined protein kinase C (PKC)-zeta/lambda activity in plasma membranes and microsomes and (b) immunoreactive PKC-zeta and PKC-lambda in GLUT4 vesicles. Activity and autophosphorylation of immunoprecipitable epitope-tagged PKC-zeta and PKC-lambda were also increased by insulin in situ and phosphatidylinositol 3,4,5-(PO(4))(3) (PIP(3)) in vitro. Because phosphoinositide dependent kinase-1 (PDK-1) is required for phosphorylation of activation loops of PKC-zeta and protein kinase B, we compared their activation. Both RO 31-8220 and myristoylated PKC-zeta pseudosubstrate blocked insulin-induced activation and autophosphorylation of PKC-zeta/lambda but did not inhibit PDK-1-dependent (a) protein kinase B phosphorylation/activation or (b) threonine 410 phosphorylation in the activation loop of PKC-zeta. Also, insulin in situ and PIP(3) in vitro activated and stimulated autophosphorylation of a PKC-zeta mutant, in which threonine 410 is replaced by glutamate (but not by an inactivating alanine) and cannot be activated by PDK-1. Surprisingly, insulin activated a truncated PKC zeta that lacks the regulatory (presumably PIP(3)-binding) domain; this may reflect PIP(3) effects on PDK-1 or transphosphorylation by endogenous full-length PKC-zeta. Our findings suggest that insulin activates both PKC-zeta and PKC lambda in plasma membranes, microsomes, and GLUT4 vesicles by a mechanism requiring increases in PIP(3), PDK-1-dependent phosphorylation of activation loop sites in PKC-zeta and lambda, and subsequent autophosphorylation and/or transphosphorylation. PMID- 10464258 TI - Antimicrobial activity of a bovine hemoglobin fragment in the tick Boophilus microplus. AB - Antifungal and antibacterial activities were detected in the hemolymph and gut contents of the cattle tick, Boophilus microplus. A peptide with antibacterial activity from the tick gut contents was purified to homogeneity by reversed-phase chromatography. The molecular mass of the purified peptide was 3,205.7 Da, measured by matrix-assisted laser desorption/ionization mass spectrometry. The amino acid sequence was obtained by Edman degradation and showed that the peptide was identical to a fragment of the bovine alpha-hemoglobin. A synthetic peptide based on the sequence obtained showed characterization data identical to those of the isolated material, confirming its structure. The synthetic peptide was active in micromolar concentrations against Gram-positive bacteria and fungi. These data led us to conclude that the antibacterial activity detected in tick gut contents is the result of enzymatic processing of a host protein, hemoglobin. This activity may be used by ticks as a defense against microorganisms. PMID- 10464259 TI - Role of walker motif A of RuvB protein in promoting branch migration of holliday junctions. Walker motif a mutations affect Atp binding, Atp hydrolyzing, and DNA binding activities of Ruvb. AB - Escherichia coli RuvB protein, an ATP-dependent hexameric DNA helicase, acts together with RuvA protein to promote branch migration of Holliday junctions during homologous recombination and recombinational repair. To elucidate the role of the Walker motif A of RuvB (GXGKT; X indicates a nonconserved residue) in ATP hydrolysis and branch migration activities, we constructed four ruvB mutant genes by site-directed mutagenesis, altering the highly conserved Lys(68) and Thr(69). K68R, K68A, and T69A mutants except T69S failed to complement UV-sensitive phenotype of the ruvB strain. These three mutant proteins, when overexpressed, made the wild-type strain UV-sensitive to varying degrees. K68R, K68A, and T69A were defective in ATP hydrolysis and branch migration activities in vitro. In the presence of Mg(2+), K68R showed markedly reduced affinity for ATP, while K68A and T69A showed only mild reduction. K68A and T69A could form hexamers in the presence of Mg(2+) and ATP, while K68R failed to form hexamers and existed instead as a higher oligomer, probably a dodecamer. In contrast to wild-type RuvB, K68R, K68A, and T69A by themselves were defective in DNA binding. However, RuvA could facilitate binding of K68A and T69A to DNA, whereas it could not promote binding of K68R to DNA. All of the three mutant RuvBs could physically interact with RuvA. These results indicate the direct involvement in ATP binding and ATP hydrolysis of the invariant Lys(68) and Thr(69) residues of Walker motif A of RuvB and suggest that these residues play key roles in interrelating these activities with the conformational change of RuvB, which is required for the branch migration activity. PMID- 10464260 TI - Stat protein transactivation domains recruit p300/CBP through widely divergent sequences. AB - The signal transduction and activator of transcription (Stat) gene family has been highly conserved throughout evolution. Gene duplication and divergence has produced 7 mammalian Stat genes, each of which mediates a distinct process. While some Stat proteins are activated by multiple cytokines, Stat2 is highly specific for responses to type I interferon. We have cloned mouse Stat2 and found that while its sequence was more divergent from its human homologue than any other mouse-human Stat pairs, it was fully functional even in human cells. Overall sequence identity was only 69%, compared with 85-99% similarity for other Stat genes, and several individual domains that still served similar or identical functions in both species were even less well conserved. The coiled-coil domain responsible for interaction with IRF9 was only 65% identical and yet mouse Stat2 interacted with either human or mouse IRF9; the carboxyl terminus was only 30% identical and yet both regions functioned as equal transactivation domains. Both mouse and human transactivation domains recruited the p300/CBP coactivator and were equally sensitive to inhibition by adenovirus E1A protein. Interestingly, the Stat3 carboxyl terminus also functioned as a transactivator capable of recruiting p300/CBP, as does the Stat1 protein, although with widely differing potencies. Yet these proteins share no sequence similarity with Stat2. These data demonstrate that highly diverged primary sequences can serve similar or identical functions, and that the minimal regions of similarity between human and mouse Stat2 may define the critical determinants for function. PMID- 10464261 TI - Interaction of non-competitive blockers within the gamma-aminobutyric acid type A chloride channel using chemically reactive probes as chemical sensors for cysteine mutants. AB - Selected channel-lining cysteine mutants from the M2 segment of rat alpha1 gamma aminobutyric acid (GABA) type A receptor subunit, at positions 257, 261, 264, and 272 were co-expressed with beta1 and gamma2 subunits in Xenopus oocytes. They generated functional receptors displaying conductance and response to both GABA and picrotoxinin similar to the wild type alpha1beta1gamma2 receptor. Three chemically reactive affinity probes derived from non-competitive blockers were synthesized to react with the engineered cysteines: 1) dithiane bis-sulfone derivative modified by an isothiocyanate function (probe A); 2) fiprole derivatives modified by an alpha-chloroketone (probe B) and alpha-bromoketone (probe C) moiety. These probes blocked the GABA-induced currents on all receptors. This blockade could be fully reversed by a washing procedure on the wild type, the alpha1T261Cbeta1gamma2 and alpha1L264Cbeta1gamma2 mutant receptors. In contrast, an irreversible effect was observed for all three probes on both alpha1V257Cbeta1gamma2 and alpha1S272Cbeta1gamma2 mutant receptors. This effect was probe concentration-dependent and could be abolished by picrotoxinin and/or t-butyl bicyclophosphorothionate. These data indicate a major interaction of non-competitive blockers at position 257 of the presumed M2 segment of rat alpha1 subunit but also suggest an interaction at the more extracellular position 272. PMID- 10464262 TI - Phosphorylation is required for alteration of kv1.5 K(+) channel function by the Kvbeta1.3 subunit. AB - The Kv1.5 K(+) channel is functionally altered by coassembly with the Kvbeta1.3 subunit, which induces fast inactivation and a hyperpolarizing shift in the activation curve. Here we examine kinase regulation of Kv1.5/Kvbeta1.3 interaction after coexpression in human embryonic kidney 293 cells. The protein kinase C inhibitor calphostin C (3 microM) removed the fast inactivation (66 +/- 1.9 versus 11 +/- 0.25%, steady state/peak current) and the beta-induced hyperpolarizing voltage shift in the activation midpoint (V(1/2)) (-21.9 +/- 1.4 versus -4.3 +/- 2.0 mV). Calphostin C had no effect on Kv1.5 alone with respect to inactivation kinetics and V(1/2). Okadaic acid, but not the inactive derivative, blunted both calphostin C effects (V(1/2) = -17.6 +/- 2.2 mV, 38 +/- 1.8% inactivation), consistent with dephosphorylation being required for calphostin C action. Calphostin C also removed the fast inactivation (57 +/- 2.6 versus 16 +/- 0.6%) and the shift in V(1/2) (-22.1 +/- 1.4 versus -2.1 +/- 2.0 mV) conferred onto Kv1.5 by the Kvbeta1.2 subunit, which shares only C terminus sequence identity with Kvbeta1. 3. In contrast, modulation of Kv1.5 by the Kvbeta2.1 subunit was unaffected by calphostin C. These data suggest that Kvbeta1.2 and Kvbeta1.3 subunit modification of Kv1.5 inactivation and voltage sensitivity require phosphorylation by protein kinase C or a related kinase. PMID- 10464263 TI - Cloning and characterization of a close homologue of human UDP-N-acetyl-alpha-D galactosamine:Polypeptide N-acetylgalactosaminyltransferase-T3, designated GalNAc T6. Evidence for genetic but not functional redundancy. AB - The UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase, designated GalNAc T3, exhibits unique functions. Specific acceptor substrates are used by GalNAc-T3 and not by other GalNAc-transferases. The expression pattern of GalNAc-T3 is restricted, and loss of expression is a characteristic feature of poorly differentiated pancreatic tumors. In the present study, a sixth human UDP GalNAc:polypeptide N-acetylgalactosaminyltransferase, designated GalNAc-T6, with high similarity to GalNAc-T3, was characterized. GalNAc-T6 exhibited high sequence similarity to GalNAc-T3 throughout the coding region, in contrast to the limited similarity that exists between homologous glycosyltransferase genes, which is usually restricted to the putative catalytic domain. The genomic organizations of GALNT3 and GALNT6 are identical with the coding regions placed in 10 exons, but the genes are localized differently at 2q31 and 12q13, respectively. Acceptor substrate specificities of GalNAc-T3 and -T6 were similar and different from other GalNAc-transferases. Northern analysis revealed distinct expression patterns, which were confirmed by immunocytology using monoclonal antibodies. In contrast to GalNAc-T3, GalNAc-T6 was expressed in WI38 fibroblast cells, indicating that GalNAc-T6 represents a candidate for synthesis of oncofetal fibronectin. The results demonstrate the existence of genetic redundancy of a polypeptide GalNAc-transferase that does not provide full functional redundancy. PMID- 10464264 TI - A novel host/tumor cell interaction activates matrix metalloproteinase 1 and mediates invasion through type I collagen. AB - Along with degradation of type IV collagen in basement membrane, destruction of the stromal collagens, types I and III, is an essential step in the invasive/metastatic behavior of tumor cells, and it is mediated, at least in part, by interstitial collagenase 1 (matrix metalloproteinase 1 (MMP-1)). Because A2058 melanoma cells produce substantial quantities of MMP-1, we used these cells as models for studying invasion of type I collagen. With a sensitive and quantitative in vitro invasion assay, we monitored the ability of these cells to invade a matrix of type I collagen and the ability of a serine proteinase inhibitor and all-trans-retinoic acid to block invasion. Although these cells produce copious amounts of MMP-1, they do not invade collagen unless they are co cultured with fibroblasts or with conditioned medium derived from fibroblasts. Our studies indicate that a proteolytic cascade that depends on stromal/tumor cell interactions facilitates the ability of A2058 melanoma cells to invade a matrix of type I collagen. This cascade activates latent MMP-1 and involves both serine proteinases and MMPs, particularly stromelysin 1 (MMP-3). All-trans retinoic acid (10(-6) M) suppresses the invasion of tumor cells by several mechanisms that include suppression of MMP synthesis and an increase in levels of tissue inhibitor of metalloproteinases 1 and 2. We conclude that invasion of stromal collagen by A2058 melanoma cells is mediated by a novel host/tumor cell interaction in which a proteolytic cascade culminates in the activation of pro MMP-1 and tumor cell invasion. PMID- 10464265 TI - Regulation of human thioredoxin reductase expression and activity by 3' untranslated region selenocysteine insertion sequence and mRNA instability elements. AB - Thioredoxin reductases function in regulating cellular redox and function through their substrate, thioredoxin, in the proper folding of enzymes and redox regulation of transcription factor activity. These enzymes are overexpressed in certain tumors and cancer cells and down-regulated in apoptosis and may play a role in regulating cell growth. Mammalian thioredoxin reductases contain a selenocysteine residue, encoded by a UGA codon, as the penultimate carboxyl terminal amino acid. This amino acid has been proposed to carry reducing equivalents from the active site to substrates. We report expression of a wild type thioredoxin reductase selenoenzyme, a cysteine mutant enzyme, and the UGA terminated protein in mammalian cells and overexpression of the cysteine mutant and UGA-terminated proteins in the baculovirus insect cell system. We show that substitution of cysteine for selenocysteine decreases enzyme activity for thioredoxin by 2 orders magnitude, and that termination at the UGA codon abolishes activity. We further demonstrate the presence of a functional selenocysteine insertion sequence element that is highly active but only moderately responsive to selenium supplementation. Finally, we show that thioredoxin reductase mRNA levels are down-regulated by other sequences in the 3' untranslated region, which contains multiple AU-rich instability elements. These sequences are found in a number of cytokine and proto-oncogene mRNAs and have been shown to confer rapid mRNA turnover. PMID- 10464266 TI - NADH shuttle system regulates K(ATP) channel-dependent pathway and steps distal to cytosolic Ca(2+) concentration elevation in glucose-induced insulin secretion. AB - The NADH shuttle system is composed of the glycerol phosphate and malate aspartate shuttles. We generated mice that lack mitochondrial glycerol-3 phosphate dehydrogenase (mGPDH), a rate-limiting enzyme of the glycerol phosphate shuttle. Application of aminooxyacetate, an inhibitor of the malate-aspartate shuttle, to mGPDH-deficient islets demonstrated that the NADH shuttle system was essential for coupling glycolysis with activation of mitochondrial ATP generation to trigger glucose-induced insulin secretion. The present study revealed that blocking the NADH shuttle system severely suppressed closure of the ATP-sensitive potassium (K(ATP)) channel and depolarization of the plasma membrane in response to glucose in beta cells, although properties of the K(ATP) channel on the excised beta cell membrane were unaffected. In mGPDH-deficient islets treated with aminooxyacetate, Ca(2+) influx through the plasma membrane induced by a depolarizing concentration of KCl in the presence of the K(ATP) channel opener diazoxide restored insulin secretion. However, the level of the secretion was only approximately 40% of wild-type controls. Thus, glucose metabolism through the NADH shuttle system leading to efficient ATP generation is pivotal to activation of both the K(ATP) channel-dependent pathway and steps distal to an elevation of cytosolic Ca(2+) concentration in glucose-induced insulin secretion. PMID- 10464267 TI - Interaction of vanadate with the cloned beta cell K(ATP) channel. AB - Vanadate is used as a tool to trap magnesium nucleotides in the catalytic site of ATPases. However, it has also been reported to activate ATP-sensitive potassium (K(ATP)) channels in the absence of nucleotides. K(ATP) channels comprise Kir6.2 and sulfonylurea receptor subunits (SUR1 in pancreatic beta cells, SUR2A in cardiac and skeletal muscle, and SUR2B in smooth muscle). We explored the effect of vanadate (2 mM), in the absence and presence of magnesium nucleotides, on different types of cloned K(ATP) channels expressed in Xenopus oocytes. Currents were recorded from inside-out patches. Vanadate inhibited Kir6.2/SUR1 currents by approximately 50% but rapidly activated Kir6.2/SUR2A ( approximately 4-fold) and Kir6. 2/SUR2B ( approximately 2-fold) currents. Mutations in SUR that abolish channel activation by magnesium nucleotides did not prevent the effects of vanadate. Studies with chimeric SUR indicate that the first six transmembrane domains account for the difference in both the kinetics and the vanadate response of Kir6.2/SUR1 and Kir6. 2/SUR2A. Boiling the vanadate solution, which removes the decavanadate polymers, largely abolished both stimulatory and inhibitory actions of vanadate. Our results demonstrate that decavanadate modulates K(ATP) channel activity via the SUR subunit, that this modulation varies with the type of SUR, that it differs from that produced by magnesium nucleotides, and that it involves transmembrane domains 1-6 of SUR. PMID- 10464268 TI - Purification of Mlc and analysis of its effects on the pts expression in Escherichia coli. AB - Products of the pts operon of Escherichia coli have multiple physiological roles such as sugar transport, and the operon is controlled by two promoters, P0 and P1. Expression of the pts P0 promoter that is increased during growth in the presence of glucose is also activated by cAMP receptor protein.cAMP. Based on the existence of a sequence that has a high similarity with the known Mlc binding site in the promoter, the effects of the Mlc protein on the pts P0 promoter expression were studied. In vivo transcription assays using wild type and mlc negative E. coli strains grown in the presence and absence of glucose indicate that Mlc negatively regulates expression of the P0 promoter, and Mlc-dependent repression is relieved by glucose in the growth medium. In vitro transcription assay using purified recombinant Mlc showed that Mlc repressed transcription from the P0 but did not affect the activity of the P1. DNase I footprinting experiments revealed that a Mlc binding site was located around +1 to +25 of the promoter and that Mlc inhibited the binding of RNA polymerase to the P0 promoter. Cells overexpressing Mlc showed a very slow fermentation rate compared with the wild type when grown in the presence of various phosphoenolpyruvate-carbohydrate phosphotransferase system sugars but few differences in the presence of non phosphoenolpyruvate-carbohydrate phosphotransferase system sugars except maltose. These results suggest that the pts operon is one of major targets for the negative regulation by Mlc, and thus Mlc regulates the utilization of various sugars as well as glucose in E. coli. The possibility that the inducer of Mlc may not be sugar or its derivative but an unknown factor is proposed to explain the Mlc induction mechanism by various sugars. PMID- 10464269 TI - Asymmetric contributions to RNA binding by the Thr(45) residues of the MS2 coat protein dimer. AB - A prominent feature of the interaction of MS2 coat protein with RNA is the quasi symmetric insertion of a bulged adenine (A-10) and a loop adenine (A-4) into conserved pockets on each subunit of the coat protein dimer. Because of its presence in both of these adenine-binding pockets, Thr(45) is thought to play an important role in interaction with RNA on both subunits of the dimer. To test the significance of Thr(45), we introduced all 19 amino acid substitutions. However, we were initially unable to determine the effects of the mutations on RNA binding because every substitution compromised the ability of coat protein to fold correctly. Genetic fusion of coat protein subunits reverted these protein structural defects, allowing us to show that the RNA binding activity of coat protein tolerates substitution of Thr(45), but only on one or the other subunit of the dimer. Single-chain heterodimer complementation experiments suggest that the primary site of Thr(45) interaction with RNA is with A-4 in the translational operator. Either contact of Thr(45) with A-10 makes little contribution to stability of the RNA-protein complex, or the effects of Thr(45) substitution are offset by conformational adjustments that introduce new, favorable contacts at nearby sites. PMID- 10464270 TI - MARCKS-related protein (MRP) is a substrate for the Leishmania major surface protease leishmanolysin (gp63). AB - Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP; MacMARCKS) are protein kinase C substrates in diverse cell types. Activation of murine macrophages by cytokines increases MRP expression, but infection with Leishmania promastigotes during activation results in MRP depletion. We therefore examined the effect of Leishmania major LV39 on recombinant MRP. Both live promastigotes and a soluble fraction of LV39 lysates degraded MRP to yield lower molecular weight fragments. Degradation was independent of MRP myristoylation and was inhibited by protein kinase C-dependent phosphorylation of MRP. MRP was similarly degraded by purified leishmanolysin (gp63), a Leishmania surface metalloprotease. Degradation was evident at low enzyme/substrate ratios, over a broad pH range, and was inhibited by 1,10 phenanthroline and by a hydroxamate dipeptide inhibitor of leishmanolysin. Using mass spectrometric analysis, cleavage was shown to occur within the effector domain of MRP between Ser(92) and Phe(93), in accordance with the substrate specificity of leishmanolysin. Moreover, an MRP construct in which the effector domain had been deleted was resistant to cleavage. Thus, Leishmania infection may result in leishmanolysin-dependent hydrolysis of MRP, a major protein kinase C substrate in macrophages. PMID- 10464271 TI - Mutations that increase acidity enhance the transcriptional activity of the glutamine-rich activation domain in stage-specific activator protein. AB - Sea urchin stage-specific activator protein (SSAP) activates transcription of the late H1 gene at the mid-blastula stage of development. Its C-terminal 202 amino acids form a potent glycine/glutamine rich activation domain (GQ domain) that can transactivate reporter genes to levels 5-fold higher than VP16 in several mammalian cell lines. We observed that, unlike other glutamine-rich activation domains, the GQ domain activates transcription to moderate levels in yeast. We utilized this activity to screen in yeast for intragenic mutations that enhance or inhibit the transcriptional activity of the GQ domain. We identified 37 loss of function and 23 gain of function mutants. Most gain of function mutations increased the acidity of the domain. The most frequently isolated mutations conferred enhanced transcriptional activity when assayed in mammalian cells. These mutations also enhance the ability of SSAP to up-regulate the late H1 promoter in sea urchin embryos. We conclude that the GQ domain fundamentally differs from other glutamine-rich activators and may share some properties of acidic activators. The ability of acidity to enhance SSAP-mediated transcription may reflect a mechanism by which phosphorylation of SSAP activates late H1 gene transcription during embryogenesis. PMID- 10464272 TI - The rate of internalization of the gonadotropin receptors is greatly affected by the origin of the extracellular domain. AB - Previous results from this laboratory have shown that human kidney (293) cells transfected with the rat follitropin receptor (rFSHR) internalize agonist (i.e. human follitropin, hFSH) at a rate similar to that of other agonist-G protein coupled receptor complexes while 293 cells transfected with the rat lutropin/choriogonadotropin receptor (rLHR) internalize agonist (human choriogonadotropin, hCG) at a rate that is about 1 order of magnitude slower. Taking advantage of this difference and the high degree of homology between the rLHR and rFSHR, we have now used chimeras of these two receptors to begin to delineate structural features that influence their internalization. Analysis of six chimeras that exchanged only the transmembrane domains (designated FLF and LFL), only the COOH-terminal domains (FFL or LLF) or both domains (FLL or LFF) show that the origin of the extracellular domain is at least as important, if not more, than the origin of the transmembrane and COOH-terminal domains in determining the rate of internalization of the gonadotropin receptors. Thus, the rates of internalization of agonist internalization mediated by FFL, FLF, and FLL more closely resemble rFSHR than rLHR, while the rates of agonist internalization mediated by LLF, LFL, and LFF more closely resemble rLHR than rFSHR. The importance of the extracellular domain was also evident even upon overexpression of arrestin-3, a protein that enhances the rate of internalization of the wild type receptors and chimeras by binding to their intracellular regions. PMID- 10464273 TI - Inflammatory cytokine regulation of Fas-mediated apoptosis in thyroid follicular cells. AB - The occurrence of apoptosis in thyroid follicular cells induced by Fas activation has been a subject of much debate. This is due, in part, to the fact that no physiologically relevant treatment conditions have been reported to cause rapid and extensive Fas-mediated apoptosis in thyroid cells, whereas treatment with the protein synthesis inhibitor cycloheximide prior to Fas activation allows for massive cell death. This indicates that the Fas signaling pathway is present but that its function is blocked in the overwhelming majority of cultured thyroid cells. To reconcile the conflicting reports, we set out to identify physiologically relevant conditions in which rapid, massive thyroid cell apoptosis in response to Fas activation could be demonstrated. We determined that susceptibility to Fas-activated apoptosis could be influenced by certain combinations of inflammatory cytokines. Although no single cytokine was effective, pretreatment of thyroid cells with the combination of gamma-interferon and either tumor necrosis factor-alpha or interleukin 1beta allowed for massive Fas-mediated apoptosis. Susceptibility to Fas-induced death correlated with an increase in expression of a tunicamycin-inhibitable high molecular weight form of Fas but not with aggregate expression of Fas. PMID- 10464274 TI - Modulation of neuronal voltage-gated calcium channels by farnesol. AB - The modulation of presynaptic voltage-dependent calcium channels by classical second messenger molecules such as protein kinase C and G protein betagamma subunits is well established and considered a key factor for the regulation of neurotransmitter release. However, little is known of other endogenous mechanisms that control the activity of these channels. Here, we demonstrate a unique modulation of N-type calcium channels by farnesol, a dephosphorylated intermediate of the mammalian mevalonate pathway. At micromolar concentrations, farnesol acts as a relatively non-discriminatory rapid open channel blocker of all types of high voltage-activated calcium channels, with a mild specificity for L-type channels. However, at 250 nM, farnesol induces an N-type channel-specific hyperpolarizing shift in channel availability that results in approximately 50% inhibition at a typical neuronal resting potential. Additional experiments demonstrated the presence of farnesol in the brain (rodents and humans) at physiologically relevant concentrations (100-800 pmol/g (wet weight)). Altogether, our results indicate that farnesol is a selective, high affinity inhibitor of N-type Ca(2+) channels and raise the possibility that endogenous farnesol and the mevalonate pathway are implicated in neurotransmitter release through regulation of presynaptic voltage-gated Ca(2+) channels. PMID- 10464275 TI - Purification, redox sensitivity, and RNA binding properties of SECIS-binding protein 2, a protein involved in selenoprotein biosynthesis. AB - In mammalian selenoprotein mRNAs, the highly structured 3' UTR contains selenocysteine insertion sequence (SECIS) elements that are required for the recognition of UGA as the selenocysteine codon. Our previous work demonstrated a tight correlation between codon-specific translational read-through and the activity of a 120-kDa RNA-binding protein that interacted specifically with the SECIS element in the phospholipid hydroperoxide glutathione peroxidase mRNA. This study reports the RNA binding and biochemical properties of this protein, SECIS binding protein 2 (SBP2). We detected SBP2 binding activity in liver, hepatoma cell, and testis extracts from which SBP2 has been purified by anion exchange and RNA affinity chromatography. This scheme has allowed us to identify a 120-kDa polypeptide that co-elutes with SBP2 binding activity from wild-type but not mutant RNA affinity columns. A characterization of SBP2 biochemical properties reveals that SBP2 binding is sensitive to oxidation and the presence of heparin, rRNA, and poly(G). SBP2 activity elutes with a molecular mass of approximately 500 kDa during gel filtration chromatography, suggesting the existence of a large functional complex. Direct cross-linking and competition experiments demonstrate that the minimal phospholipid hydroperoxide glutathione peroxidase 3' UTR binding site is between 82 and 102 nucleotides, which correlates with the minimal sequence necessary for translational read-through. SBP2 also interacts specifically with the minimally functional 3' UTR of another selenoprotein mRNA, deiodinase 1. PMID- 10464276 TI - Heparin is a unique marker of progenitors in the glial cell lineage. AB - The oligodendrocyte-type-2 astrocyte progenitor cells (precursors of oligodendrocytes and type-2 astrocytes) are an excellent system in which to study differentiation as they can be manipulated in vitro. Maintenance of oligodendrocyte-type-2 astrocyte progenitor cells requires basic fibroblast growth factor, a growth factor whose action normally depends on a heparan sulfate coreceptor. Biochemical analysis revealed a most surprising result: that the oligodendrocyte-type-2 astrocyte progenitors did not synthesize heparan sulfate, the near ubiquitous N-sulfated cell surface polysaccharide, but the chemically related heparin in a form that was almost completely N- and O-sulfated. The heparin was detected in the pericellular fraction of the cells and the culture medium. In contrast the differentiated glial subpopulations (oligodendrocytes and type-2 astrocytes) synthesized typical heparan sulfate but with distinctive fine structural features for each cell type. Thus heparin is a unique differentiation marker in the glial lineage. Previously heparin has been found only in a subset of mature mast cells called the connective tissue mast cells. Its presence within the developing nervous system on a precise population of progenitors may confer specific and essential recognition properties on those cells in relation to binding soluble growth and/or differentiation factors and the extracellular matrix. PMID- 10464277 TI - Helical interactions and membrane disposition of the 16-kDa proteolipid subunit of the vacuolar H(+)-ATPase analyzed by cysteine replacement mutagenesis. AB - Theoretical mechanisms of proton translocation by the vacuolar H(+)-ATPase require that a transmembrane acidic residue of the multicopy 16-kDa proteolipid subunit be exposed at the exterior surface of the membrane sector of the enzyme, contacting the lipid phase. However, structural support for this theoretical mechanism is lacking. To address this, we have used cysteine mutagenesis to produce a molecular model of the 16-kDa proteolipid complex. Transmembrane helical contacts were determined using oxidative cysteine cross-linking, and accessibility of cysteines to the lipid phase was determined by their reactivity to the lipid-soluble probe N-(1-pyrenyl)maleimide. A single model for organization of the four helices of each monomeric proteolipid was the best fit to the experimental data, with helix 1 lining a central pore and helix 2 and helix 3 immediately external to it and forming the principal intermolecular contacts. Helix 4, containing the crucial acidic residue, is peripheral to the complex. The model is consistent not only with theoretical proton transport mechanisms, but has structural similarity to the dodecameric ring complex formed by the related 8-kDa proteolipid of the F(1)F(0)-ATPase. This suggests some commonality between the proton translocating mechanisms of the vacuolar and F(1)F(0)-ATPases. PMID- 10464278 TI - Commitment of neutrophilic differentiation and proliferation of HL-60 cells coincides with expression of transferrin receptor. Effect of granulocyte colony stimulating factor on differentiation and proliferation. AB - To examine the regulatory mechanisms of proliferation and maturation in neutrophilic lineage cells, we have tried to sort dimethyl sulfoxide (Me(2)SO) treated HL-60 cells into transferrin receptor (Trf-R) positive (Trf-R(+)) and negative (Trf-R(-)) cells. Differentiated Trf-R(-) cells expressed more formyl Met-Leu-Phe receptor (fMLP-receptor) and ability of O-(2) genaration, as markers of differentiation, than Trf-R(+) cells, and Trf-R(-) cell differentiation was markedly accelerated by the incubation with granulocyte colony stimulating factor (G-CSF). On the other hand, Trf-R(+) cells had a tendency to proliferate rather than differentiate, and proliferation was enhanced by G-CSF. These results indicate that Trf-R expression coincides with the commitment to proliferate or differentiate of HL-60 cells, and G-CSF accelerates these commitments. G-CSF induced tyrosine phosphorylation of STAT 3 in Trf-R(-) cells much more than in Trf-R(+) cells. Protein 70 S6 kinase expression was higher in Trf-R(+) cells than in Trf-R(-) cells. Furthermore, p70 S6 kinase was hyperphosphorylated by G-CSF in Trf-R(+) cells, but not in Trf-R(-) cells. Rapamycin, an inhibitor of p70 S6 kinase activity, inhibited G-CSF-dependent proliferation of Trf-R(+) cells and increased fMLP-R expression on these cells. These results suggest that commitment to proliferation and differentiation in Me(2)SO-treated HL-60 cells is preprogrammed and correlated with Trf-R expression, and G-CSF potentiates the cellular commitment. STAT 3 may promote differentiation of Me(2)SO-treated HL-60 cells into neutrophils, while p70 S6 kinase may promote proliferation and negatively regulate neutrophilic differentiation. PMID- 10464279 TI - alpha-synuclein binds to Tau and stimulates the protein kinase A-catalyzed tau phosphorylation of serine residues 262 and 356. AB - alpha-Synuclein has been implicated in the pathogenesis of several neurodegenerative disorders based on the direct linking of missense mutations in alpha-synuclein to autosomal dominant Parkinson's disease and its presence in Lewy-like lesions. To gain insight into alpha-synuclein functions, we have investigated whether it binds neuronal proteins and modulates their functional state. The microtubule-associated protein tau was identified as a ligand by alpha synuclein affinity chromatography of human brain cytosol. Direct binding assays using (125)I-labeled human tau40 demonstrated a reversible binding with a IC(50) about 50 pM. The interacting domains were localized to the C terminus of alpha synuclein and the microtubule binding region of tau as determined by protein fragmentation and the use of recombinant peptides. High concentrations of tubulin inhibited the binding between tau and alpha-synuclein. Functionally, alpha synuclein stimulated the protein kinase A-catalyzed phosphorylation of tau serine residues 262 and 356 as determined using a phospho-epitope-specific antibody. We propose that alpha-synuclein modulates the phosphorylation of soluble axonal tau and thereby indirectly affects the stability of axonal microtubules. PMID- 10464280 TI - Molecular interactions among protein phosphatase 2A, tau, and microtubules. Implications for the regulation of tau phosphorylation and the development of tauopathies. AB - Hyperphosphorylated forms of the neuronal microtubule (MT)-associated protein tau are major components of Alzheimer's disease paired helical filaments. Previously, we reported that ABalphaC, the dominant brain isoform of protein phosphatase 2A (PP2A), is localized on MTs, binds directly to tau, and is a major tau phosphatase in cells. We now describe direct interactions among tau, PP2A, and MTs at the submolecular level. Using tau deletion mutants, we found that ABalphaC binds a domain on tau that is indistinguishable from its MT-binding domain. ABalphaC binds directly to MTs through a site that encompasses its catalytic subunit and is distinct from its binding site for tau, and ABalphaC and tau bind to different domains on MTs. Specific PP2A isoforms bind to MTs with distinct affinities in vitro, and these interactions differentially inhibit the ability of PP2A to dephosphorylate various substrates, including tau and tubulin. Finally, tubulin assembly decreases PP2A activity in vitro, suggesting that PP2A activity can be modulated by MT dynamics in vivo. Taken together, these findings indicate how structural interactions among ABalphaC, tau, and MTs might control the phosphorylation state of tau. Disruption of these normal interactions could contribute significantly to development of tauopathies such as Alzheimer's disease. PMID- 10464281 TI - Cellular physiology of STAT3: Where's the cytoplasmic monomer? AB - In the standard model of cytokine-induced signal transducer and activator of transcription (STAT) protein family signaling to the cell nucleus, it is assumed that STAT3 is recruited to the cytoplasmic side of the cell surface receptor complex from within a cytosolic monomer pool. By using Superose-6 gel-filtration chromatography, we have discovered that there is little monomeric STAT3 (91 kDa) in the cytosol of liver cells (human hepatoma Hep3B cell line and rat liver). The bulk of STAT3 (and STAT1, STAT5a, and -b) was present in the cytosol as high molecular mass complexes in two broad distributions in the size range 200-400 kDa ("statosome I") and 1-2 MDa ("statosome II"). Upon treatment of Hep3B cells with interleukin-6 (IL-6) for 30 min (i) cytosolic tyrosine-phosphorylated STAT3 was found to be in complexes of size ranging from 200-400 kDa to 1-2 MDa; (ii) a small pool of monomeric STAT3 and tyrosine-phosphorylated STAT3 eluting at 80-100 kDa was observed, and (iii) most of the cytoplasmic DNA-binding competent STAT3 (the so-called SIF-A "homodimer") co-eluted with catalase at 230 kDa. In order to identify the protein components of the 200-400-kDa statosome I cytosolic complexes, we used the novel technique of antibody-subtracted differential protein display using anti-STAT3 antibody. Eight polypeptides in the size range from 20 to 114 kDa co-shifted with STAT3; three of these (p60, p20a, and p20b) were co-shifted in an IL-6-dependent manner. In-gel tryptic fragmentation and mass spectroscopy identified the major IL-6-dependent STAT3-co-shifted p60 protein as the chaperone GRP58/ER-60/ERp57. Taken together, these data (i) emphasize the absence of a detectable STAT3 monomer pool in the cytosol of cytokine-free liver cells as posited by the standard model, and (ii) suggest an alternative model for STAT signaling in which STAT3 proteins function in the cytoplasm as heteromeric complexes with accessory scaffolding proteins, including the chaperone GRP58. PMID- 10464282 TI - Thrombin interacts with thrombomodulin, protein C, and thrombin-activatable fibrinolysis inhibitor via specific and distinct domains. AB - A collection of 56 purified thrombin mutants, in which 76 charged or polar surface residues on thrombin were mutated to alanine, was used to identify key residues mediating the interactions of thrombin with thrombomodulin (TM), protein C, and thrombin-activatable fibrinolysis inhibitor (TAFI). Comparison of protein C activation in the presence and absence of TM identified 11 residues mediating the thrombin-TM interaction (Lys(21), Gln(24), Arg(62), Lys(65), His(66), Arg(68), Thr(69), Tyr(71), Arg(73), Lys(77), Lys(106)). Three mutants (E25A, D51A, R89A/R93A/E94A) were found to have decreased ability to activate TAFI yet retained normal protein C activation, whereas three other mutants (R178A/R180A/D183A, E229A, R233A) had decreased ability to activate protein C but maintained normal TAFI activation. One mutant (W50A) displayed decreased activation of both substrates. Mapping of these functional residues on thrombin revealed that the 11 residues mediating the thrombin-TM interaction are all located in exosite I. Residues important in TAFI activation are located above the active-site cleft, whereas residues involved in protein C are located below the active-site cleft. In contrast to the extensive overlap of residues mediating TM binding and fibrinogen clotting, these data show that distinct domains in thrombin mediate its interactions with TM, protein C, and TAFI. These studies demonstrate that selective enzymatic properties of thrombin can be dissociated by site-directed mutagenesis. PMID- 10464283 TI - Rab11BP/Rabphilin-11, a downstream target of rab11 small G protein implicated in vesicle recycling. AB - Rab11 small G protein has been implicated in vesicle recycling, but its upstream regulators or downstream targets have not yet been identified. We isolated here a downstream target of Rab11, named rabphilin-11, from bovine brain. Moreover, we isolated from a rat brain cDNA library its cDNA, which encoded a protein with a M(r) of 100,946 and 908 amino acids (aa). Rabphilin-11 bound GTP-Rab11 more preferentially than GDP-Rab11 at the N-terminal region and was specific for Rab11 and inactive for other Rab and Rho small G proteins. Both GTP-Rab11 and rabphilin 11 were colocalized at perinuclear regions, presumably the Golgi complex and recycling endosomes, in Madin-Darby canine kidney cells. In HeLa cells cultured on fibronectin, both the proteins were localized not only at perinuclear regions but also along microtubules, which were oriented toward membrane lamellipodia. Treatment of HeLa cells with nocodazole caused disruption of microtubules and dispersion of GTP-Rab11 and rabphilin-11. Overexpression of the C-terminal fragment of rabphilin-11 (aa 607-730), lacking the GTP-Rab11 binding domain, in HeLa cells reduced accumulation of transferrin at perinuclear regions and cell migration. Rabphilin-11 turned out to be a rat counterpart of recently reported bovine Rab11BP. These results indicate that rabphilin-11 is a downstream target of Rab11 which is involved in vesicle recycling. PMID- 10464284 TI - Regulated CD44 cleavage under the control of protein kinase C, calcium influx, and the Rho family of small G proteins. AB - CD44 is a cell surface receptor for several extracellular matrix components and is implicated in tumor cell invasion and metastasis. Our previous studies have shown that CD44 expressed in cancer cells is proteolytically cleaved at the extracellular domain through membrane-associated metalloproteases and that CD44 cleavage plays a critical role in CD44-mediated tumor cell migration (Okamoto, I., Kawano, Y., Tsuiki, H., Sasaki, J., Nakao, M., Matsumoto, M., Suga, M., Ando, M., Nakajima, M., and Saya, H. (1999) Oncogene 18, 1435-1446). In the present study, we first demonstrate rapid degradation of the membrane-tethered CD44 cleavage product through intracellular proteolytic pathways, and it occurs only after CD44 extracellular cleavage. To address the mechanisms regulating CD44 cleavage at the extracellular domain, we show that 12-O-tetradecanoylphorbol 13 acetate (TPA) and the calcium ionophore ionomycin rapidly enhance metalloprotease mediated CD44 cleavage in U251MG cells via protein kinase C-dependent and independent pathways, respectively, suggesting the existence of multiple distinct pathways for regulation of CD44 cleavage. Concomitant with TPA-induced CD44 cleavage, TPA treatment induces redistribution of CD44 and ERM proteins (ezrin, radixin, and moesin) to newly generated membrane ruffling areas. Treatment with lysophosphatidic acid, which is known to activate the Rho-dependent pathway, inhibits TPA-induced CD44 redistribution and CD44 cleavage. Furthermore, overexpression of Rac dominant active mutants results in the redistribution of CD44 to the Rac-induced ruffling areas and the enhancement of CD44 cleavage. These results suggest that the Rho family proteins play a role in regulation of CD44 distribution and cleavage. PMID- 10464285 TI - The nature of the catalytic domain of 2'-5'-oligoadenylate synthetases. AB - 2'-5'-Oligoadenylate (2-5(A)) synthetases are a family of interferon-induced enzymes that are activated by double-stranded RNA. To understand why, unlike other DNA and RNA polymerases, they catalyze 2'-5' instead of 3'-5' phosphodiester bond formation, we used molecular modeling to compare the structure of the catalytic domain of DNA polymerase beta (pol beta) to that of a region of the P69 isozyme of 2-5(A) synthetase. Although the primary sequence identity is low, like pol beta, P69 can assume an alphabetabetaalphabetabetabeta structure in this region. Moreover, mutation of the three Asp residues of P69, which correspond to the three catalytic site Asp residues of pol beta, inactivated the enzyme without affecting its substrate and activator binding capacity, providing further credence to the concept that this region is the catalytic domain of P69. This domain is highly conserved among all 2-5(A) synthetase isozymes. Biochemical and mutational studies demonstrated that dimerization of the P69 protein is required for its enzyme activity. However, a dimer containing a wild type subunit and an inactive catalytic domain mutant subunit was also active. The rate of catalysis of the heterodimer was half of that of the wild type homodimer, although the two proteins bound double-stranded RNA and ATP equally well. PMID- 10464286 TI - Identification of a novel phosphorylation site on histone H3 coupled with mitotic chromosome condensation. AB - Histone H3 (H3) phosphorylation at Ser(10) occurs during mitosis in eukaryotes and was recently shown to play an important role in chromosome condensation in Tetrahymena. When producing monoclonal antibodies that recognize glial fibrillary acidic protein phosphorylation at Thr(7), we obtained some monoclonal antibodies that cross-reacted with early mitotic chromosomes. They reacted with 15-kDa phosphoprotein specifically in mitotic cell lysate. With microsequencing, this phosphoprotein was proved to be H3. Mutational analysis revealed that they recognized H3 Ser(28) phosphorylation. Then we produced a monoclonal antibody, HTA28, using a phosphopeptide corresponding to phosphorylated H3 Ser(28). This antibody specifically recognized the phosphorylation of H3 Ser(28) but not that of glial fibrillary acidic protein Thr(7). Immunocytochemical studies with HTA28 revealed that Ser(28) phosphorylation occurred in chromosomes predominantly during early mitosis and coincided with the initiation of mitotic chromosome condensation. Biochemical analyses using (32)P-labeled mitotic cells also confirmed that H3 is phosphorylated at Ser(28) during early mitosis. In addition, we found that H3 is phosphorylated at Ser(28) as well as Ser(10) when premature chromosome condensation was induced in tsBN2 cells. These observations suggest that H3 phosphorylation at Ser(28), together with Ser(10), is a conserved event and is likely to be involved in mitotic chromosome condensation. PMID- 10464287 TI - Additive effects of beta chain mutations in low oxygen affinity hemoglobin betaF41Y,K66T. AB - In order to decrease significantly the oxygen affinity of human hemoglobin, we have associated the mutation betaF41Y with another point mutation also known to decrease the oxygen affinity of Hb. We have synthesized a recombinant Hb (rHb) with two mutations in the beta chains: rHb betaF41Y,K66T. In the absence of 2, 3 diphosphoglycerate, additive effects of the mutations are evident, since the doubly mutated Hb exhibits a larger decrease in oxygen affinity than for the individual single mutations. In the presence of 2,3-diphosphoglycerate, the second mutation did not significantly increase the P(50) value relative to the single mutations. However, the kinetics of CO binding still indicate combined effects on the allosteric equilibrium, as evidenced by more of the slow bimolecular phase characteristic of binding to the deoxy conformation. Dimer tetramer equilibrium studies indicate an increase in stability of the mutants relative to rHb A; the double mutant rHb betaF41Y, K66T at pH 7.5 showed a K(4,2) value of 0.26 microM. Despite the lower oxygen affinity, the single mutant betaF41Y and double mutant betaF41Y,K66T show only a moderate increase of 20% in the autoxidation rate. These mutations are thus of interest in developing a Hb based blood substitute. PMID- 10464288 TI - ADAM-TS5, ADAM-TS6, and ADAM-TS7, novel members of a new family of zinc metalloproteases. General features and genomic distribution of the ADAM-TS family. AB - We report the primary structure of three novel, putative zinc metalloproteases designated ADAM-TS5, ADAM-TS6, and ADAM-TS7. All have a similar domain organization, comprising a preproregion, a reprolysin-type catalytic domain, a disintegrin-like domain, a thrombospondin type-1 (TS) module, a cysteine-rich domain, a spacer domain without cysteine residues, and a COOH-terminal TS module. These genes are differentially regulated during mouse embryogenesis and in adult tissues, with Adamts5 highly expressed in the peri-implantation period in embryo and trophoblast. These proteins are similar to four other cognate gene products, defining a distinct family of human reprolysin-like metalloproteases, the ADAM-TS family. The other members of the family are ADAM-TS1, an inflammation-induced gene, the procollagen I/II amino-propeptide processing enzyme (PCINP, ADAM-TS2), and proteins predicted by the KIAA0366 and KIAA0688 genes (ADAM-TS3 and ADAM TS4). Individual ADAM-TS members differ in the number of COOH-terminal TS modules, and some have unique COOH-terminal domains. The ADAM-TS genes are dispersed in human and mouse genomes. PMID- 10464290 TI - Recruitment of ATM protein to double strand DNA irradiated with ionizing radiation. AB - The product of the ATM gene, which is mutated in ataxia telangiectasia, is a nuclear phosphoprotein, and it involves the activation of the p53 pathway after ionizing radiation. Here we show that the ATM protein is constitutively associated with double strand DNA and that the interaction increases when the DNA is exposed to ionizing radiation. The ATM protein also had affinity to restriction endonuclease PvuII-digested DNA, but not to UV-irradiated DNA nor X irradiated single-stranded DNA. The immunoprecipitation experiment detected very weak association between ATM and DNA-PK proteins, and immunodepletion of DNA-PK showed little or no effect on the interaction of the ATM protein with damaged DNA, indicating that an interaction with DNA-PK might not be required for the recruitment of the ATM protein to damaged DNA. Furthermore, the association was also confirmed in xrs-5 and xrs-6e cells, which are Chinese hamster ovary mutant cell lines defective in Ku80 function. These results indicate that the ATM protein is recruited to the site of DNA damage and it recognizes double strand breaks by itself or through an association with other DNA-binding protein other than DNA-PK and Ku80 proteins. PMID- 10464289 TI - Mapping the substrate binding site of the prostaglandin transporter PGT by cysteine scanning mutagenesis. AB - We have identified a cDNA, PGT, that encodes a widely expressed transporter for prostaglandin (PG) E(2), PGF(2alpha), PGD(2), 8-iso-PGF(2alpha), and thromboxane B(2). To begin to understand the molecular mechanisms of transporter function, we have initiated a structure-function analysis of PGT to identify its substrate binding region. We have found that by introducing the small, water-soluble, thiol reactive anion Na(2-sulfonatoethyl)methanethiosulfonate (MTSES) into the substrate pathway, we were able to cause inhibition of transport that could be reversed with dithiothreitol. Importantly, co-incubation with PGE(2) protected PGT from this inhibition, suggesting that MTSES gains access to the aqueous pore pathway of PGT to form a mixed disulfide near the substrate-binding site. To identify the susceptible cysteine, we mutated, one at a time, all six of the putative transmembrane cysteines to serine. Only the mutation of Cys-530 to serine within putative transmembrane 10 became resistant to inhibition by MTSES. Thus, Cys-530 is the substrate-protectable, MTSES-inhibitable residue. To identify other residues that may be lining the substrate-binding site, we initiated cysteine-scanning mutagenesis of transmembrane 10 using Cys-530 as an entry point. On a C530S, MTSES-resistant background, residues in the N- and C terminal directions were individually mutated to cysteine (Ala-513 to His-536), one at a time, and then analyzed for MTSES inhibition. Of the 24 cysteine substituted mutants generated, 6 were MTSES-sensitive and, among these, 4 were substrate-protectable. The pattern of sensitivity to MTSES places these residues on the same face of an alpha-helix. The results of cysteine-scanning mutagenesis and molecular modeling of putative transmembrane 10 indicate that the substrate binding of PGT is formed among its membrane-spanning segments, with 4 residues along the cytoplasmic end of helix 10 contributing to one surface of the binding site. PMID- 10464291 TI - Transcriptional regulation of the platelet-derived growth factor alpha receptor gene via CCAAT/enhancer-binding protein-delta in vascular smooth muscle cells. AB - Inflammatory cytokines stimulate the proliferation of vascular smooth muscle cells (VSMC) and play a pivotal role in the pathogenesis of vascular diseases including atherosclerosis and restenosis. Mitogenic response of interleukin-1beta (IL-1beta) on VSMC is thought to be mediated by induction of endogenous platelet derived growth factor (PDGF), especially PDGF-AA. Although the action of PDGF-AA is mediated by its specific receptor, PDGFalpha-receptor (PDGFalphaR), very little is known about the regulatory mechanism of PDGFalphaR gene expression in VSMC. To understand the mechanism, we studied the transcriptional control of the PDGFalphaR gene in VSMC after treatment with IL-1beta. IL-1beta (10 ng/ml) drastically increased both PDGFalphaR and CCAAT/enhancer-binding protein delta (C/EBPdelta) mRNA levels in a time dependent manner. A rapid induction of C/EBPdelta mRNA within 30 min was followed by slower emergence of PDGFalphaR mRNA, which reached the maximum level in 12 h, whereas C/EBPdelta mRNA was detectable at 30 min and reached the maximum level at 3 h. Electromobility shift and supershift assays revealed that IL-1beta markedly increased DNA-protein complex, which was mainly composed of C/EBPbeta and/or -delta. Both Western blotting and immunohistochemistry demonstrated that either C/EBPbeta or -delta expression was induced by IL-1beta exclusively in nuclei of VSMC. On the other hand, overexpression of C/EBPdelta specifically transactivated the promoter activity of the PDGFalphaR gene and significantly enhanced VSMC proliferation in PDGF-treated cells. We conclude that induction of PDGFalphaR expression is mainly mediated by C/EBPdelta expression in VSMC, and a high level of C/EBPdelta expression may be involved in the pathogenesis of atherosclerosis and restenosis. PMID- 10464292 TI - Dimerization of guanylyl cyclase-activating protein and a mechanism of photoreceptor guanylyl cyclase activation. AB - Ca(2+)-binding guanylyl cyclase-activating proteins (GCAPs) stimulate photoreceptor membrane guanylyl cyclase (retGC) in the light when the free Ca(2+) concentrations in photoreceptors decrease from 600 to 50 nM. RetGC activated by GCAPs exhibits tight dimerization revealed by chemical cross-linking (Yu, H., Olshevskaya, E., Duda, T., Seno, K., Hayashi, F., Sharma, R. K., Dizhoor, A. M., and Yamazaki, A. (1999) J. Biol. Chem. 274, 15547-15555). We have found that the Ca(2+)-loaded GCAP-2 monomer undergoes reversible dimerization upon dissociation of Ca(2+). The ability of GCAP-2 and its several mutants to activate retGC in vitro correlates with their ability to dimerize at low free Ca(2+) concentrations. A constitutively active GCAP-2 mutant E80Q/E116Q/D158N that stimulates retGC regardless of the free Ca(2+) concentrations forms dimers both in the absence and in the presence of Ca(2+). Several GCAP-2/neurocalcin chimera proteins that cannot efficiently activate retGC in low Ca(2+) concentrations are also unable to dimerize in the absence of Ca(2+). Additional mutation that restores normal activity of the GCAP-2 chimera mutant also restores its ability to dimerize in the absence of Ca(2+). These results suggest that dimerization of GCAP-2 can be a part of the mechanism by which GCAP-2 regulates the photoreceptor guanylyl cyclase. The Ca(2+)-free GCAP-1 is also capable of dimerization in the absence of Ca(2+), but unlike GCAP-2, dimerization of GCAP-1 is resistant to the presence of Ca(2+). PMID- 10464293 TI - Two separate cis-active elements of the vasoactive intestinal peptide gene mediate constitutive and inducible transcription by binding different sets of AP 1 proteins. AB - Vasoactive intestinal peptide (VIP) gene expression is highly restricted throughout the neuroaxis and regulated by extracellular factors that activate tyrosine- or serine/threonine-directed protein kinase pathways. Cytokine, cyclic AMP, and tissue-specific response elements on the VIP gene have been characterized. Those mediating responsiveness to protein kinase C have not. The endogenous VIP gene and a 5.2-kilobase pair (kb) VIP-luciferase reporter gene, are up-regulated by phorbol 12-myristate 13-acetate (PMA) in SK-N-SH neuroblastoma cells. PMA stimulation was abolished by deletion of sequences at 1.37 to -1.28 or -1.28 to -0.904 kb, but not by removal of the single phorbol ester response element (TRE; TGACTCA) located at -2.25 kb. Mutation of sites at 1.32 or -1.20 that mediate neurotrophin responsiveness of the VIP gene (Symes, A., Lewis, S., Corpus, L., Rajan, P., Hyman, S. E., and Fink, J. S. (1994) Mol. Endocrinol. 8, 1750-1763) each reduced PMA induction in SK-N-SH cells by >50%, and double mutation abolished it. The two mutations also reduced basal VIP reporter gene transcription in SH-EP neuroblastoma cells expressing VIP constitutively. Both cis-active elements bound pre-existing AP-1 proteins in SH EP- or PMA-stimulated SK-N-SH cell nuclear extracts. The AP-1 complex at both sites contained a Fos-related protein with c-Jun in SH-EP cells and c-Fos with a Jun-related protein in SK-N-SH cells. Recruitment of combinatorially distinct AP 1 complexes to these elements may underlie cell type-specific regulation of the VIP gene. PMID- 10464294 TI - Differential localization of 5- and 15-lipoxygenases to the nuclear envelope in RAW macrophages. AB - Leukotriene formation is initiated in myeloid cells by an increase in intracellular calcium and translocation of 5-lipoxygenase from the cytoplasm to the nuclear envelope where it can utilize arachidonic acid. Monocyte- macrophages and eosinophils also express 15-lipoxygenase, which converts arachidonic acid to 15(S)-hydroxyeicosatetraenoic acid. Enhanced green fluorescent 5-lipoxygenase (5 LO) and 15-lipoxygenase (15-LO) fusion proteins were expressed in the cytoplasm of RAW 264.7 macrophages. Only 5-lipoxygenase translocated to the nuclear envelope after cell stimulation, suggesting that differential subcellular compartmentalization can regulate the generation of leukotrienes versus 15(S) hydroxyeicosatetraenoic acid in cells that possess both lipoxygenases. A series of truncation mutants of 5-LO were created to identify putative targeting domains; none of these mutants localized to the nuclear envelope. The lack of targeting of 15-LO was then exploited to search for specific targeting motifs in 5-LO, by creating 5-LO/15-LO chimeric molecules. The only chimera that could sustain nuclear envelope translocation was one which involved replacement of the N-terminal 237 amino acids with the corresponding segment of 15-LO. Significantly, no discrete targeting domain could be identified in 5-LO, suggesting that sequences throughout the molecule are required for nuclear envelope localization. PMID- 10464295 TI - Mode analysis of a fatty acid molecule binding to the N-terminal 8-kDa domain of DNA polymerase beta. A 1:1 complex and binding surface. AB - We reported previously that long-chain fatty acids are potent inhibitors of mammalian DNA polymerase beta. At present, based on information available from the NMR structure of the N-terminal 8-kDa domain, we examined the structural interaction with the 8-kDa domain using two species, C(18)-linoleic acid (LA) or C(24)-nervonic acid (NA). In the 8-kDa domain with LA or NA, the structure that forms the interaction interface included helix-1, helix-2, helix-4, the three turns (residues 1-13, 48-51, and 79-87) and residues adjacent to an Omega-type loop connecting helix-1 and helix-2 of the same face. No significant shifts were observed for any of the residues on the opposite side of the 8-kDa domain. The NA interaction interface on the amino acid residues of the 8-kDa domain fragment was mostly the same as that of LA, except that the shifted cross-peaks of Leu-11 and Thr-79 were significantly changed between LA and NA. The 8-kDa domain bound to LA or NA as a 1:1 complex with a dissociation constant (K(D)) of 1.02 or 2.64 mM, respectively. PMID- 10464296 TI - Structure and function of HNK-1 sulfotransferase. Identification of donor and acceptor binding sites by site-directed mutagenesis. AB - HNK-1 glycan, sulfo-->3GlcAbeta1-->3Galbeta1-->4GlcNAc-->R, is uniquely enriched in neural cells and natural killer cells and is thought to play important roles in cell-cell interaction. HNK-1 glycan synthesis is dependent on HNK-1 sulfotransferase (HNK-1ST), and cDNAs encoding human and rat HNK-1ST have been recently cloned. HNK-1ST belongs to the sulfotransferase gene family, which shares two homologous sequences in their catalytic domains. In the present study, we have individually mutated amino acid residues in these conserved sequences and determined how such mutations affect the binding to the donor substrate, adenosine 3'-phosphate 5'-phosphosulfate, and an acceptor. Mutations of Lys(128), Arg(189), Asp(190), Pro(191), and Ser(197) to Ala all abolished the enzymatic activity. When Lys(128) and Asp(190) were conservatively mutated to Arg and Glu, respectively, however, the mutated enzymes still maintained residual activity, and both mutant enzymes still bound to adenosine 3',5'-diphosphate-agarose. K128R and D190E mutant enzymes, on the other hand, exhibited reduced affinity to the acceptor as demonstrated by kinetic studies. These findings, together with those on the crystal structure of estrogen sulfotransferase and heparan sulfate N deacetylase/sulfotransferase, suggest that Lys(128) may be close to the 3 hydroxyl group of beta-glucuronic acid in a HNK-1 acceptor. In contrast, the effect by mutation at Asp(190) may be due to conformational change because this amino acid and Pro(191) reside in a transition of the secondary structure of the enzyme. These results indicate that conserved amino acid residues in HNK-1ST play roles in maintaining a functional conformation and are directly involved in binding to donor and acceptor substrates. PMID- 10464297 TI - The high resolution crystal structure of recombinant Crithidia fasciculata tryparedoxin-I. AB - Tryparedoxin-I is a recently discovered thiol-disulfide oxidoreductase involved in the regulation of oxidative stress in parasitic trypanosomatids. The crystal structure of recombinant Crithidia fasciculata tryparedoxin-I in the oxidized state has been determined using multi-wavelength anomalous dispersion methods applied to a selenomethionyl derivative. The model comprises residues 3 to 145 with 236 water molecules and has been refined using all data between a 19- and 1.4-A resolution to an R-factor and R-free of 19.1 and 22.3%, respectively. Despite sharing only about 20% sequence identity, tryparedoxin-I presents a five stranded twisted beta-sheet and two elements of helical structure in the same type of fold as displayed by thioredoxin, the archetypal thiol-disulfide oxidoreductase. However, the relationship of secondary structure with the linear amino acid sequences is different for each protein, producing a distinctive topology. The beta-sheet core is extended in the trypanosomatid protein with an N terminal beta-hairpin. There are also differences in the content and orientation of helical elements of secondary structure positioned at the surface of the proteins, which leads to different shapes and charge distributions between human thioredoxin and tryparedoxin-I. A right-handed redox-active disulfide is formed between Cys-40 and Cys-43 at the N-terminal region of a distorted alpha-helix (alpha1). Cys-40 is solvent-accessible, and Cys-43 is positioned in a hydrophilic cavity. Three C-H...O hydrogen bonds donated from two proline residues serve to stabilize the disulfide-carrying helix and support the correct alignment of active site residues. The accurate model for tryparedoxin-I allows for comparisons with the family of thiol-disulfide oxidoreductases and provides a template for the discovery or design of selective inhibitors of hydroperoxide metabolism in trypanosomes. Such inhibitors are sought as potential therapies against a range of human pathogens. PMID- 10464298 TI - Lysosomal and cytosolic sialic acid 9-O-acetylesterase activities can Be encoded by one gene via differential usage of a signal peptide-encoding exon at the N terminus. AB - 9-O-Acetylation is one of the most common modifications of sialic acids, and it can affect several sialic acid-mediated recognition phenomena. We previously reported a cDNA encoding a lysosomal sialic acid-specific 9-O-acetylesterase, which traverses the endoplasmic reticulum-Golgi pathway and localizes primarily to lysosomes and endosomes. In this study, we report a variant cDNA derived from the same gene that contains a different 5' region. This cDNA has a putative open reading frame lacking a signal peptide-encoding sequence and is thus a candidate for the previously described cytosolic sialic acid 9-O-acetylesterase activity. Epitope-tagged constructs confirm that the new sequence causes the protein product to be targeted to the cytosol and has esterase activity. Using reverse transcription-polymerase chain reaction to distinguish the two forms of message, we show that although the lysosomal sialic acid-specific 9-O-acetylesterase message has a widespread pattern of expression in adult mouse tissues, this cytosolic sialic acid 9-O-acetylesterase form has a rather restricted distribution, with the strongest expression in the liver, ovary, and brain. Using a polyclonal antibody directed against the 69-amino acid region common to both proteins, we confirmed that the expression of glycosylated and nonglycosylated polypeptides occurred in appropriate subcellular fractions of normal mouse tissues. Rodent liver polypeptides reacting to the antibody also co-purify with previously described lysosomal sialic acid esterase activity and at least a portion of the cytosolic activity. Thus, two sialic acid 9-O-acetylesterases found in very different subcellular compartments can be encoded by a single gene by differential usage of a signal peptide-encoding exon at the N terminus. The 5' rapid amplification of cDNA ends results and the differences in tissue-specific expression suggest that expression of these two products may be differentially regulated by independent promoters. PMID- 10464299 TI - Phenotypic behavior of caveolin-3 mutations that cause autosomal dominant limb girdle muscular dystrophy (LGMD-1C). Retention of LGMD-1C caveolin-3 mutants within the golgi complex. AB - Caveolin-3, a muscle-specific caveolin-related protein, is the principal structural protein of caveolae membrane domains in striated muscle cell types (cardiac and skeletal). Autosomal dominant limb girdle muscular dystrophy (LGMD 1C) in humans is due to mutations within the caveolin-3 gene: (i) a 9-base pair microdeletion that removes three amino acids within the caveolin scaffolding domain (DeltaTFT) or (ii) a missense mutation within the membrane spanning domain (P --> L). The molecular mechanisms by which these two mutations cause muscular dystrophy remain unknown. Here, we investigate the phenotypic behavior of these caveolin-3 mutations using heterologous expression. Wild type caveolin-3 or caveolin-3 mutants were transiently expressed in NIH 3T3 cells. LGMD-1C mutants of caveolin-3 (DeltaTFT or P --> L) were primarily retained at the level of a perinuclear compartment that we identified as the Golgi complex in double labeling experiments, while wild type caveolin-3 was efficiently targeted to the plasma membrane. In accordance with these observations, caveolin-3 mutants formed oligomers of a much larger size than wild type caveolin-3 and were excluded from caveolae-enriched membrane fractions as seen by sucrose density gradient centrifugation. In addition, these caveolin-3 mutants were expressed at significantly lower levels and had a dramatically shortened half-life of approximately 45-60 min. However, caveolin-3 mutants were palmitoylated to the same extent as wild type caveolin-3, indicating that targeting to the plasma membrane is not required for palmitoylation of caveolin-3. In conclusion, we show that LGMD-1C mutations lead to formation of unstable high molecular mass aggregates of caveolin-3 that are retained within the Golgi complex and are not targeted to the plasma membrane. Consistent with its autosomal dominant form of genetic transmission, we demonstrate that LGMD-1C mutants of caveolin-3 behave in a dominant-negative fashion, causing the retention of wild type caveolin-3 at the level of the Golgi. These data provide a molecular explanation for why caveolin-3 levels are down-regulated in patients with this form of limb girdle muscular dystrophy (LGMD-1C). PMID- 10464300 TI - RNA editing associated with the generation of two distinct conformations of the trypanosomatid Leptomonas collosoma 7SL RNA. AB - Analysis of the trypanosomatid Leptomonas collosoma 7SL RNA revealed the existence of two distinct stable 7SL RNA conformers (7SL I and II). Sequence analysis of the RNAs indicated a single base difference between the conformers at position 133 (C in 7SL II and U in 7SL I) located in domain III. This change appears to be the result of a post-transcriptional editing event, since the single-copy 7SL RNA gene codes exclusively for a C at this position. The edited form (7SL I) was found preferentially in the cytoplasm, and the pre-edited form in the nucleus. 7SL I is mainly bound to ribosomes, whereas 7SL II is more abundant in ribosome-free particles. Mutations introduced in regions outside the editing site were found to occur in a single conformation, suggesting that the editing event is not the only factor that determines the conformation of the molecule. This study is the first description of an editing event on a small RNA other than tRNA and is the first report of C --> U editing in trypanosomes. We propose a novel role for RNA editing in controlling the conformation of the 7SL RNA in vivo. PMID- 10464301 TI - The p185(neu)-containing glycoprotein complex of a microfilament-associated signal transduction particle. Purification, reconstitution, and molecular associations with p58(gag) and actin. AB - Microfilaments associate with the microvillar membrane of 13762 ascites mammary adenocarcinoma cells via a large transmembrane complex (TMC) comprising the major glycoproteins TMC-gp120, -110, -80, -65, and -55, the receptor kinase p185(neu), and the cytoplasmic proteins actin and p58(gag), linking the receptor with microfilaments in a signal transduction particle. Immunoblot screening with polyclonal antisera to TMC glycoproteins showed selective epithelial expression in normal rat tissues and epithelially derived tumor cells. The TMC glycoproteins were isolated by solubilization of microfilament core preparations in SDS, dilution, and separation on a concanavalin A-agarose affinity column. The large p185(neu)-containing complex was reconstituted from the column eluate after displacement of SDS with nonionic detergent, demonstrated by gel filtration and co-immunoprecipitation of the glycoproteins with anti-gp55 or anti-p185(neu). Exhaustive biotinylation of the glycoproteins gave a stoichiometry of gp120:gp110:gp80:gp65:gp55 of approximately 1:1:1:0.5:1. Overlay blots with biotinylated actin and in vitro translated, [(35)S]methionine-labeled p58(gag), respectively, showed specific interactions of actin with gp55 and gp120 and of p58(gag) with gp65 and gp55. These results provide evidence for a specific complex of microfilament-associated glycoproteins containing p185(neu) and p58(gag) and suggest a role for the complex in signal transduction scaffolding. PMID- 10464302 TI - Association of the Ras to mitogen-activated protein kinase signal transduction pathway with microfilaments. Evidence for a p185(neu)-containing cell surface signal transduction particle linking the mitogenic pathway to a membrane microfilament association site. AB - Microvilli of the aggressive 13762 ascites mammary adenocarcinoma contain a large, microfilament-associated signal transduction particle whose scaffolding is a stable glycoprotein complex (Li, Y., Hua, F., Carraway, K. L., and Carraway, C. A. C. (1999) J. Biol. Chem. 274, 25651-25658) associated with the growth factor receptor p185(neu). The receptor is constitutively tyrosine-phosphorylated in the cells and microvilli, predicting that it should recruit mitogenic pathway components to this membrane-microfilament interaction site. Immunoprecipitation of cell lysates with anti-phosphotyrosine and immunoblotting showed phosphorylated forms of the mitogenic pathway proteins Shc and MAPK in addition to p185(neu), suggesting that the Ras to MAPK mitogenic pathway is activated. Immunoblotting of p185(neu)-containing microvillar fractions revealed the presence in each of stably associated Shc, Grb-2, Sos, Ras, Raf, mitogen activated protein kinase kinase, and mitogen-activated protein kinase/extracellular signal-regulated kinase, as well as the transcription factor phosphorylating kinase Rsk. All of these pathway components co-immunoprecipitated with p185(neu) from cleared lysates of microvilli solubilized under microfilament depolymerizing conditions. The recruitment of constitutively phosphorylated p185(neu) and the activated mitogenic pathway proteins to this membrane microfilament interaction site provides a physical model for integrating the assembly of the mitogenic pathway with the transmission of growth factor signal to the cytoskeleton. This linkage is probably a requisite step in the global cytoskeleton remodeling accompanying mitogenesis. PMID- 10464303 TI - Evidence for a novel cardiac-enriched retinoid X receptor partner. AB - Recent studies indicate that retinoid-mediated pathways play a pivotal role in cardiac morphogenesis and function. To identify proteins that serve as interacting partners of the retinoid X receptor alpha (RXRalpha) in heart, DNA protein binding studies were performed with an RXR-responsive element (NRRE-1) derived from the medium chain acyl-CoA dehydrogenase gene promoter and nuclear protein extracts prepared from adult rat heart. NRRE-1 is a pleiotropic RXR responsive element comprised of three potential recognition sites for class II members of the nuclear receptor superfamily. Gel mobility shift assays performed with an NRRE-1 probe in the absence or presence of bacterially overproduced RXRalpha and nuclear protein extracts prepared from adult rat heart, liver, or brain identified a cardiac-specific, RXR-dependent DNA-protein interaction. The NRRE-1-RXR.cardiac-enriched RXR-interacting protein (CERIP) complex exhibited a distinct mobility compared with NRRE-1-RXR.peroxisome proliferator-activated receptor, NRRE-1-RXR.retinoic acid receptor, or NRRE-1-RXR.thyroid receptor complexes. Mutational analysis demonstrated that two of the three potential binding half-sites of NRRE-1 (an everted repeat separated by an 8-base pair spacer) are required for the NRRE-1-RXR. CERIP interaction. Gel mobility shift assays demonstrated that CERIP interacted with RXRalpha and RXRgamma but not with RXRbeta, indicating a receptor subtypespecific binding preference and suggesting an RXR AB region-dependent interaction. The RXR.CERIP complex did not form on NRRE-1 when a mutant GST-RXRalpha fusion protein lacking the NH(2)-terminal AB region (but containing the receptor dimerization domain) of RXRalpha was added in place of the full-length RXRalpha, confirming a role for the AB region in the RXR. CERIP interaction. DNA-protein cross-linking studies demonstrated that CERIP is a DNA-binding protein of approximately 110 kDa. These results provide evidence for the existence of a cardiac-enriched DNA-binding protein that interacts with RXRalpha via the AB region and suggest a mechanism whereby cardiac retinoid signaling is controlled in an RXR subtype-specific manner. PMID- 10464304 TI - Cell-free expression and functional reconstitution of homo-oligomeric alpha7 nicotinic acetylcholine receptors into planar lipid bilayers. AB - The alpha7 nicotinic acetylcholine receptor (nAChR) is a ligand-gated ion channel that modulates neurotransmitter release in the central nervous system. We show here that functional, homo-oligomeric alpha7 nAChRs can be synthesized in vitro with a rabbit reticulocyte lysate translation system supplemented with endoplasmic reticulum microsomes, reconstituted into planar lipid bilayers, and evaluated using single-channel recording techniques. Because wild-type alpha7 nAChRs desensitize rapidly, we used a nondesensitizing form of the alpha7 receptor with mutations in the second transmembrane domain (S2'T and L9'T) to record channel activity in the continuous presence of agonist. Endoglycosidase H treatment of microsomes containing nascent alpha7 S2'T/L9'T nAChRs indicated that the receptors were glycosylated. A proteinase K protection assay revealed a 36 kDa fragment in the ER lumen, consistent with a large extracellular domain predicted by most topological models, indicating that the protein was folded integrally through the ER membrane. alpha7 S2'T/L9'T receptors reconstituted into planar lipid bilayers had a unitary conductance of approximately 50 pS, were highly selective for monovalent cations over Cl(-), were nonselective between K(+) and Na(+), and were blocked by alpha-bungarotoxin. This is the first demonstration that a functional ligand-gated ion channel can be synthesized using an in vitro expression system. PMID- 10464305 TI - Stable association of 70-kDa heat shock protein induces latent multisite specificity of a unisite-specific endonuclease in yeast mitochondria. AB - The multisite-specific endonuclease Endo.SceI of yeast mitochondria is unique among endonucleases because its 50-kDa subunit forms a stable dimer with the mitochondrial 70-kDa heat shock protein (mtHSP70), which otherwise fulfills a chaperone function by binding transiently to unfolded proteins. Here we show that the mtHSP70 subunit confers broader sequence specificity, greater stability, and higher activity on the 50-kDa subunit. The 50-kDa subunit alone displayed weaker activity and highly sequence-specific endonuclease activity. The 50-kDa protein exists as a heterodimer with mtHSP70 in vivo, allowing Endo.SceI to cleave specifically at multiple sites on mitochondrial DNA. Endo.SceI may have evolved from a highly specific endonuclease that gained broader sequence specificity after becoming a stable partner of mtHSP70. PMID- 10464306 TI - A 220-kDa activator complex of the 26 S proteasome in insects and humans. A role in type II programmed insect muscle cell death and cross-activation of proteasomes from different species. AB - The S10b (SUG2) ATPase cDNA has been cloned by reverse transcription-polymerase chain reaction/rapid amplification of cDNA ends from mRNA of intersegmental muscles of the tobacco horn moth (Manduca sexta). The S10b ATPase is a component of the 26 S proteasome, and its concentration and that of its mRNA increase dramatically during development in a manner similar to other ATPases of the 19 S regulator of the 26 S proteasome. The S10b and S6' (TBP1) ATPases are also present in a complex of approximately 220 kDa in intersegmental muscles. The 220 kDa complex markedly activates (2-10-fold) the 26 S proteasome, even when bound to anti-S10b antibodies immobilized on Sepharose, and increases in concentration approximately 5-fold like the 26 S proteasome in the intersegmental muscles in preparation for the programmed death of the muscle cells. A similar activator complex is present in human brain and placenta. Free activator complexes cross activate: the Manduca complex activates rat skeletal muscle 26 S proteasomes, and the placental complex activates Manduca 26 S proteasomes. The placental activator complex contains S10b and S6', but not p27. This 220-kDa activator complex has been evolutionarily conserved between species from insect to man and may have a fundamental role in proteasome regulation. PMID- 10464307 TI - Regulation of AMP deaminase by phosphoinositides. AB - AMP deaminase (AMPD) converts AMP to IMP and is a diverse and highly regulated enzyme that is a key component of the adenylate catabolic pathway. In this report, we identify the high affinity interaction between AMPD and phosphoinositides as a mechanism for regulation of this enzyme. We demonstrate that endogenous rat brain AMPD and the human AMPD3 recombinant enzymes specifically bind inositide-based affinity probes and to mixed lipid micelles that contain phosphatidylinositol 4,5-bisphosphate. Moreover, we show that phosphoinositides specifically inhibit AMPD catalytic activity. Phosphatidylinositol 4,5-bisphosphate is the most potent inhibitor, effecting pure noncompetitive inhibition of the wild type human AMPD3 recombinant enzyme with a K(i) of 110 nM. AMPD activity can be released from membrane fractions by in vitro treatment with neomycin, a phosphoinositide-binding drug. In addition, in vivo modulation of phosphoinositide levels leads to a change in the soluble and membrane-associated pools of AMPD activity. The predicted human AMPD3 sequence contains pleckstrin homology domains and (R/K)X(n)(R/K)XKK sequences, both of which are characterized phosphoinositide-binding motifs. The interaction between AMPD and phosphoinositides may mediate membrane localization of the enzyme and function to modulate catalytic activity in vivo. PMID- 10464308 TI - Caveolin-2 localizes to the golgi complex but redistributes to plasma membrane, caveolae, and rafts when co-expressed with caveolin-1. AB - We have characterized comparatively the subcellular distributions of caveolins-1 and -2, their interactions and their roles in caveolar formation in polarized epithelial cells. In Fischer rat thyroid (FRT) cells, which express low levels of caveolin-2 and no caveolin-1, caveolin-2 localizes exclusively to the Golgi complex but is partially redistributed to the plasma membrane upon co-expression of caveolin-1 by transfection or by adenovirus-mediated transduction. In Madin Darby canine kidney (MDCK) cells, which constitutively express both caveolin-1 and -2, caveolin-2 localized to both the Golgi complex and to the plasma membrane, where it co-distributed with caveolin-1 in flat patches and in caveolae. In FRT cells, endogenous or overexpressed caveolin-2 did not associate with low density Triton insoluble membranes that floated in sucrose density gradients but was recruited to these membranes when co-expressed together with caveolin-1. In MDCK cells, both caveolin-1 and caveolin-2 associated with low density Triton-insoluble membranes. In FRT cells, transfection of caveolin-1 promoted the assembly of plasma membrane caveolae that localized preferentially (over 99%) to the basolateral surface, like constitutive caveolae of MDCK cells. In contrast, as expected from its intracellular distribution, endogenous or overexpressed caveolin-2 did not promote the assembly of caveolae; rather, it appeared to promote the assembly of intracellular vesicles in the peri-Golgi area. The data reported here demonstrate that caveolin-1 and -2 have different and complementary subcellular localizations and functional properties in polarized epithelial cells and suggest that the two proteins co-operate to carry out specific as yet unknown tasks between the Golgi complex and the cell surface. PMID- 10464309 TI - Expression of caveolin-1 is required for the transport of caveolin-2 to the plasma membrane. Retention of caveolin-2 at the level of the golgi complex. AB - Caveolins-1 and -2 are normally co-expressed, and they form a hetero-oligomeric complex in many cell types. These caveolin hetero-oligomers are thought to represent the assembly units that drive caveolae formation in vivo. However, the functional significance of the interaction between caveolins-1 and -2 remains unknown. Here, we show that caveolin-1 co-expression is required for the transport of caveolin-2 from the Golgi complex to the plasma membrane. We identified a human erythroleukemic cell line, K562, that expresses caveolin-2 but fails to express detectable levels of caveolin-1. This allowed us to stringently assess the effects of recombinant caveolin-1 expression on the behavior of endogenous caveolin-2. We show that expression of caveolin-1 in K562 cells is sufficient to reconstitute the de novo formation of caveolae in these cells. In addition, recombinant expression of caveolin-1 allows caveolin-2 to form high molecular mass oligomers that are targeted to caveolae-enriched membrane fractions. In striking contrast, in the absence of caveolin-1 expression, caveolin-2 forms low molecular mass oligomers that are retained at the level of the Golgi complex. Interestingly, we also show that expression of caveolin-1 in K562 cells dramatically up-regulates the expression of endogenous caveolin-2. Northern blot analysis reveals that caveolin-2 mRNA levels remain constant under these conditions, suggesting that the expression of caveolin-1 stabilizes the caveolin-2 protein. Conversely, transient expression of caveolin-2 in CHO cells is sufficient to up-regulate endogenous caveolin-1 expression. Thus, the formation of a hetero-oligomeric complex between caveolins-1 and -2 stabilizes the caveolin-2 protein product and allows caveolin-2 to be transported from the Golgi complex to the plasma membrane. PMID- 10464310 TI - Fibroblast growth factor receptor-1-mediated endothelial cell proliferation is dependent on the Src homology (SH) 2/SH3 domain-containing adaptor protein Crk. AB - Stimulation of fibroblast growth factor receptor-1 (FGFR-1) expressed on endothelial cells leads to cellular migration and proliferation. We have examined the role of the Src homology (SH) 2/SH3 domain-containing adaptor protein Crk in these processes. Transient tyrosine phosphorylation of Crk in fibroblast growth factor-2-stimulated endothelial cells was dependent on the juxtamembrane tyrosine residue 463 in FGFR-1, and a Crk SH2 domain precipitated FGFR-1 via phosphorylated Tyr-463, indicating direct complex formation between Crk and FGFR 1. Furthermore, Crk SH2 and SH3 domains formed ligand-independent complexes with Shc, C3G, and the Crk-associated substrate (Cas). Tyrosine phosphorylation of C3G and Cas increased as a consequence of growth factor treatment. We examined the role of Crk in FGFR-1-mediated cellular responses by use of cells expressing chimeric platelet-derived growth factor receptor-alpha/FGFR-1 (alphaR/FR) wild type and mutant Y463F receptors. The kinase activity of alphaR/FR Y463F was intact, but both Crk and the adaptor FRS-2 were no longer tyrosine-phosphorylated in the mutant cells. Both wild type and mutant receptor cells migrated efficiently, whereas cells expressing the mutant alphaR/FR Y463F failed to proliferate and Erk2 and Jun kinase activities were suppressed in these cells. In wild type alphaR/FR cells transiently expressing an SH2 domain mutant of Crk, Erk and Jun kinase activities as well as DNA synthesis were attenuated. Our data indicate that Crk participates in signaling complexes downstream of FGFR-1, which propagate mitogenic signals. PMID- 10464311 TI - cDNA cloning and chromosomal localization of human alpha(11) integrin. A collagen binding, I domain-containing, beta(1)-associated integrin alpha-chain present in muscle tissues. AB - We previously identified a novel integrin alpha-chain in human fetal muscle cells (Gullberg, D., Velling, T., Sjoberg, G., and Sejersen, T. (1995) Dev. Dyn. 204, 57-65). We have now isolated the full-length cDNA for this integrin subunit, alpha(11). The open reading frame of the cDNA encodes a precursor of 1188 amino acids. The predicted mature protein of 1166 amino acids contains seven conserved FG-GAP repeats, an I domain with a metal ion-dependent adhesion site motif, a short transmembrane region, and a unique cytoplasmic domain of 24 amino acids containing the sequence GFFRS. alpha(11), like other I domain integrins, lacks a dibasic cleavage site for generation of a heavy chain and a light chain, and it contains three potential divalent cation binding sites in repeats 5-7. The presence of 22 inserted amino acids in the extracellular stalk portion (amino acids 804-826) distinguishes the alpha(11) integrin sequence from other integrin alpha-chains. Amino acid sequence comparisons reveal the highest identity of 42% with the alpha(10) integrin chain. Immunoprecipitation with antibodies to alpha(11) integrin captures a 145-kDa protein distinctly larger than the 140-kDa alpha(2) integrin chain when analyzed by SDS-polyacrylamide gel electrophoresis under nonreducing conditions. Fluorescence in situ hybridization maps the integrin alpha(11) gene to chromosome 15q23, in the vicinity of an identified locus for Bardet-Biedl syndrome. Based on Northern blotting, integrin alpha(11) mRNA levels are high in the adult human uterus and in the heart and intermediate in skeletal muscle and some other tissues tested. During in vitro myogenic differentiation, alpha(11) mRNA and protein are up-regulated. Studies of ligand binding properties show that alpha(11)beta(1) binds collagen type I-Sepharose, and cultured muscle cells localize alpha(11)beta(1) into focal contacts on collagen type I. Future studies will reveal the importance of alpha(11)beta(1) for muscle development and integrity in adult muscle and other tissues. PMID- 10464312 TI - Evidence for a p21(ras)/Raf-1/MEK-1/ERK-2-independent pathway in stimulation of IL-2 gene transcription in human primary T lymphocytes. AB - T cell stimulation leads to triggering of signals transmitted from the cell membrane to the nucleus through TCR/CD3 proteins. Characterization of these signals largely results from the use of cell lines stimulated with anti-CD3 monoclonal antibodies. These studies have established that activation caused a rapid increase in the formation of GTP-bound Ras, which stimulates the mitogen activated protein kinase pathway involving the extracellular-regulated kinase-2 (ERK-2) and activates the nuclear factor of activated T cells (NF-AT) that regulates interleukin-2 (IL-2) gene transcription. In the present study, we used human primary T cells, and we investigated the intracellular signals triggered by two different anti-CD3 monoclonal antibodies (UCHT1 and X-35), which both strongly induce cell proliferation. We found that, in contrast to the commonly used UCHT1, X-35 activated IL-2 gene transcription without stimulation of the Raf 1/mitogen-activated ERK kinase-1 (MEK-1)/ERK-2 phosphorylation cascade; we also showed that X-35 stimulation, which triggers an ERK-2-independent pathway, does not involve activation of p21(ras). In addition to demonstrating that activation of p21(ras) and of its Raf-1/MEK-1/ERK-2 effector pathway is not an event obligatorily triggered upon TCR/CD3 ligation, these results provide the first evidence of the existence of a p21(ras)/ERK-2-independent pathway for IL-2 gene transcription in human primary T lymphocytes. PMID- 10464313 TI - Effect of p53 protein redox states on binding to supercoiled and linear DNA. AB - The binding of p53 to its DNA consensus sequence is modulated by the redox state of the protein in vitro. We have shown previously that reduced wild-type p53 binds strongly to supercoiled DNA (scDNA) regardless of the presence or absence of p53CON. Here we compare the effects of oxidation of p53 by azodicarboxylic acid bis[dimethylamide] (diamide) and other agents on p53 binding to p53CON and to scDNA. Oxidation decreases the binding of p53 to scDNA; however, under conditions where binding to p53CON in a DNA fragment is completely abolished, some residual binding to scDNA is still observed. Increasing the concentration of oxidized p53 confers minimal changes in p53 binding to both scDNA and p53CON. Reduction of the oxidized protein by dithiothreitol neither restores its binding to DNA nor to p53CON in DNA fragments. In the presence of excess zinc ions, oxidation of p53 is, however, reversible. We conclude that the irreversibility of p53 oxidation is due, at least in part, to the removal of intrinsic zinc from its position in the DNA binding domain accompanied by a conformational change of the p53 molecule after oxidation of the three cysteines to which the zinc ion is coordinated in the reduced protein. PMID- 10464314 TI - Cooperative assembly of androgen receptor into a nucleoprotein complex that regulates the prostate-specific antigen enhancer. AB - Prostate cancer is characterized by elevated serum levels of prostate-specific antigen (PSA). PSA gene expression is controlled by an androgen-responsive transcriptional enhancer. Our study suggests that formation of a nucleoprotein complex, encompassing 170 base pairs of enhancer DNA, mediates androgen responsive PSA enhancer activity. The complex is assembled by cooperative binding of androgen receptor to at least four tandem, nonconsensus androgen response elements (AREs). Systematic mutagenesis of the AREs demonstrated that they act synergistically to stimulate androgen receptor-responsive gene expression. We discuss a mechanism whereby a combination of high androgen receptor levels in the prostate and low affinity AREs contribute to the cell type specificity and activity of the enhancer. PMID- 10464315 TI - p38 mitogen-activated protein kinase is involved in Fas ligand expression. AB - p38 mitogen-activated protein kinase (MAPK) is activated by T cell receptor engagement. Here we showed that T cell receptor activated p38alpha but not p38delta. Inhibition of p38alpha by the specific inhibitor SB 203580 prevented activation-induced cell death in T cells. SB 203580 had no effect on Fas initiated apoptosis. Instead, SB 203580 preferentially inhibited activation induced Fas ligand (FasL) expression. The inhibition on FasL expression by SB 203580 was correlated with the suppression on the FasL promoter activation. Overexpression of active MAPK kinase 3b, the activator of p38 MAPK, led to activation of FasL promoter and induction of FasL transcripts in T cells. Stress stimulation of T cells by anisomycin also induced FasL expression in a p38 MAPK dependent manner. The induction of FasL expression in nonlymphoid cells such as 293T also required activation of p38 MAPK. Our results suggest that p38 MAPK is essential for FasL expression. PMID- 10464316 TI - Mapping of the discontinuous H-kininogen binding site of plasma prekallikrein. Evidence for a critical role of apple domain-2. AB - Plasma prekallikrein, a zymogen of the contact phase system, circulates in plasma as heterodimeric complex with H-kininogen. The binding is mediated by the prekallikrein heavy chain consisting of four apple domains, A1 to A4, to which H kininogen binds with high specificity and affinity (K(D) = 1.2 x 10(-8) M). Previous work had demonstrated that a discontinuous kininogen-binding site is formed by a proximal part located in A1, a distal part exposed by A4, and other yet unidentified portion(s) of the kallikrein heavy chain. To detect relevant binding segment(s) we recombinantly expressed single apple domains and found a rank order of binding affinity for kininogen of A2 > A4 approximately A1 > A3. Removal of single apple domains in prekallikrein deletion mutants reduced kininogen binding by 21 (A1), 64 (A2), and 24% (A4), respectively, whereas deletion of A3 was without effect. Transposition of homologous A2 domain from prekallikrein to factor XI conferred high-affinity kininogen binding from the former to the latter. The principal role of A2 for H-kininogen docking to the prekallikrein heavy chain was further substantiated by the finding that cleavage of a single peptide bond in A2 drastically diminished the H-kininogen binding affinity. Furthermore, the epitope of monoclonal antibody PKH6 which blocks kallikrein-kininogen complex formation with an IC(50) of 8 nM mapped to the center portion of domain A2. Our data indicate that domain A2 and two flanking sequence segments of A1 and A4 form a discontinuous binding platform for H kininogen on the prekallikrein heavy chain. Domain-specific antibodies directed to these critical sites efficiently interfered with contact phase-induced bradykinin release from H-kininogen. PMID- 10464317 TI - Coordinate regulation of the alpha(2)-macroglobulin signaling receptor and the low density lipoprotein receptor-related protein/alpha(2)-macroglobulin receptor by insulin. AB - We have studied insulin-dependent regulation of macrophage alpha(2)-macroglobulin signaling receptors (alpha(2)MSR) and low density lipoprotein receptor-related protein/alpha(2)M receptors (LRP/alpha(2)MR) employing cell binding of (125)I alpha(2)M*, inhibition of binding by receptor-associated protein (RAP) or Ni(2+), LRP/alpha(2)MR mRNA levels, and generation of second messengers. Insulin treatment increased the number of alpha(2)M* high (alpha(2)MSR) and low (LRP/alpha(2)MR) affinity binding sites from 1, 600 and 67,000 to 2,900 and 115,200 sites per cell, respectively. Neither RAP nor Ni(2+) blocked the binding of (125)I-alpha(2)M* to alpha(2)MSR on insulin- or buffer-treated cells, but they both blocked binding to LRP/alpha(2)MR. Insulin significantly increased LRP/alpha(2)MR mRNA levels in a dose- and time-dependent manner. Insulin augmented (125)I-alpha(2)M* binding to macrophages was severely reduced by wortmannin, LY294002, PD98059, SB203580, or rapamycin. The increase in alpha(2)MSR receptor synthesis was reflected by augmented generation of IP(3) and increased [Ca(2+)](i) levels upon receptor ligation. Incubation of macrophages with wortmannin, LY294002, PD98059, SB203580, rapamycin, or antibodies against insulin receptors before insulin treatment and alpha(2)M* stimulation significantly reduced the insulin-augmented increase in IP(3) and [Ca(2+)](i) levels. Pretreatment of cells with actinomycin D or cycloheximide blocked the synthesis of new alpha(2)MSR. In conclusion, we show here that insulin coordinately regulates macrophage alpha(2)MSR and LRP/alpha(2)MR, utilizing both the PI 3-kinase and Ras signaling pathways to induce new synthesis of these receptors. PMID- 10464318 TI - DNA methylation at mammalian replication origins. AB - In Escherichia coli, DNA methylation regulates both origin usage and the time required to reassemble prereplication complexes at replication origins. In mammals, at least three replication origins are associated with a high density cluster of methylated CpG dinucleotides, and others whose methylation status has not yet been characterized have the potential to exhibit a similar DNA methylation pattern. One of these origins is found within the approximately 2 kilobase pair region upstream of the human c-myc gene that contains 86 CpGs. Application of the bisulfite method for detecting 5-methylcytosines at specific DNA sequences revealed that this region was not methylated in either total genomic DNA or newly synthesized DNA. Therefore, DNA methylation is not a universal component of mammalian replication origins. To determine whether or not DNA methylation plays a role in regulating the activity of origins that are methylated, the rate of remethylation and the effect of hypomethylation were determined at origin beta (ori-beta), downstream of the hamster DHFR gene. Remethylation at ori-beta did not begin until approximately 500 base pairs of DNA was synthesized, but it was then completed by the time that 4 kilobase pairs of DNA was synthesized (<3 min after release into S phase). Thus, DNA methylation cannot play a significant role in regulating reassembly of prereplication complexes in mammalian cells, as it does in E. coli. To determine whether or not DNA methylation plays any role in origin activity, hypomethylated hamster cells were examined for ori-beta activity. Cells that were >50% reduced in methylation at ori-beta no longer selectively activated ori-beta. Therefore, at some loci, DNA methylation either directly or indirectly determines where replication begins. PMID- 10464319 TI - Different mechanisms of c-Jun NH(2)-terminal kinase-1 (JNK1) activation by ultraviolet-B radiation and by oxidative stressors. AB - Irradiation of mammalian cells with ultraviolet-B radiation (UV-B) triggers the activation of a group of stress-activated protein kinases known as c-Jun NH(2) terminal kinases (JNKs). UV-B activates JNKs via UV-B-induced ribotoxic stress. Because oxidative stress also activates JNKs, we have addressed the question of whether the ribotoxic and the oxidative stress responses are mechanistically similar. The pro-oxidants sodium arsenite, cadmium chloride, and hydrogen peroxide activated JNK1 with slow kinetics, whereas UV-B potentiated the activity of JNK1 rapidly. N-acetyl cysteine (a scavenger of reactive oxygen intermediates) abolished the ability of all oxidative stressors tested to activate JNK1, but failed to affect the activation of JNK1 by UV-B or by another ribotoxic stressor, the antibiotic anisomycin. In contrast, emetine, an inhibitor of the ribotoxic stress response, was unable to inhibit the activation of JNK1 by oxidative stressors. Although UV-A and long wavelength UV-B are the spectral components of the ultraviolet solar radiation that cause significant oxidative damage to macromolecules, the use of a filter to eliminate the radiation output from wavelengths below 310 nm abolished the activation of JNK1 by UV. Our results are consistent with the notion that UV-B and oxidative stressors trigger the activation of JNK1 through different signal transduction pathways. PMID- 10464320 TI - Evidence that transcription factor IIB is required for a post-assembly step in transcription initiation. AB - Mutation of glutamate 62 to lysine in yeast transcription factor (TF) IIB (Sua7) causes a cold-sensitive phenotype. This mutant also leads to preferential transcription of downstream start sites on some promoters in vivo. To explore the molecular nature of these phenotypes, the TFIIB E62K mutant was characterized in vitro. The mutant interacts with TATA-binding protein normally. In three different assays, the mutant can also interact with RNA polymerase II and recruit it and the other basal transcription factors to a promoter. Despite the ability to assemble a transcription complex, the TFIIB E62K protein is severely defective in transcription in vitro. Therefore, the role of TFIIB must be more than simply bridging TATA-binding protein and polymerase at the promoter. We propose that the region around Glu-62 in yeast TFIIB plays a role in start site selection, perhaps mediating a conformational change in the polymerase or the DNA during the search for initiation sites. This step may be related to the yeast-specific spacing between TATA elements and start sites since mutations of the corresponding glutamate in mammalian TFIIB do not produce a similar effect. PMID- 10464321 TI - The mouse gene PDCR encodes a peroxisomal delta(2), delta(4)-dienoyl-CoA reductase. AB - Here we describe the identification and characterization of a novel mouse gene, PDCR, that encodes a peroxisomal Delta(2), Delta(4)-dienoyl-CoA reductase. The mouse PDCR cDNA contains an 892-base pair open reading frame and is predicted to encode a 292-amino acid protein with a deduced molecular mass of 31,298 Da that terminates in a consensus type-1 peroxisomal targeting signal. Purified recombinant PDCR protein was generated from Escherichia coli and catalyzed the NADPH-dependent reduction of Delta(2)-trans, Delta(4)-trans-decadienoyl-CoA with a specific activity of 20 units/mg. Enzymatic characterization followed by high pressure liquid chromatography analysis of the products revealed that PDCR converted Delta(2)-trans,Delta(4)-trans-decadienoyl-CoA to a Delta(3)-enoyl-CoA but not to a Delta(2)-enoyl-CoA. Kinetic analyses demonstrated that PDCR is active on a broad range of Delta(2), Delta(4)-dienoyl-CoAs. Although the observed substrate preference was to Delta(2)-trans,Delta(4)-trans-decadienoyl-CoA, PDCR was also active on a C(22) substrate with multiple unsaturations, a result consistent with the role of peroxisomes in the oxidation of complex, very long chain, polyunsaturated fatty acids. The presence of a type-1 peroxisomal targeting signal Ala-Lys-Leu-COOH at the C terminus of PDCR suggested that this protein may be peroxisomal. We observed that tagged PDCR was efficiently transported to the peroxisome lumen in normal human fibroblasts but not in cells derived from a Zellweger syndrome patient with a specific defect in peroxisomal matrix protein import. We conclude that this protein resides within the peroxisome matrix and therefore represents the first mammalian peroxisomal Delta(2),Delta(4)-dienoyl-CoA reductase to be characterized at the molecular level. PMID- 10464322 TI - Nitric oxide controls src kinase activity through a sulfhydryl group modification mediated Tyr-527-independent and Tyr-416-linked mechanism. AB - c-Src kinase was activated when either murine NIH3T3 fibroblast cells or immunoprecipitated c-Src proteins were treated with nitric oxide generator, S nitroso-N-acetyl penicillamine (SNAP) or sodium nitroprusside. Nitric oxide (NO) scavenger hemoglobin and N(2)O(3) scavenger homocysteine abolished the SNAP mediated c-Src kinase activation. Phosphoamino acid analysis and peptide mapping of in vitro labeled phospho-c-Src proteins revealed that SNAP promoted the autophosphorylation at tyrosine, which preferentially took place at Tyr-416. Peptide mapping of in vivo labeled c-Src kinase excluded the involvement of phospho-Tyr-527 dephosphorylation in the SNAP-mediated activation mechanism. Correspondingly, protein-tyrosine phosphatase inhibitor Na(3)VO(4) did not abolish the SNAP-mediated activation of Src kinase, and the constitutively activated v-Src kinase was also further up-regulated in activity by SNAP. SNAP, however, failed to up-regulate the kinase activity of Phe-416 mutant v-Src. 2 Mercaptoethanol or dithiothreitol, which should disrupt N(2)O(3)-mediated S nitrosylation and subsequent formation of the S-S bond, abolished the up regulated catalytic activity, and the activity was regained after re-exposing the enzyme to SNAP. Exposure of Src kinase to SNAP promoted both autophosphorylation and S-S bond-mediated aggregation of the kinase molecules, demonstrating a linkage between the two events. These results suggest that the NO/N(2)O(3) provoked S-nitrosylation/S-S bond formation destabilizes the Src structure for Tyr-416 autophosphorylation-associated activation bypassing the Tyr-527-linked regulation. PMID- 10464323 TI - Cobalt-dependent transcriptional switching by a dual-effector MerR-like protein regulates a cobalt-exporting variant CPx-type ATPase. AB - CoaR associates with and confers cobalt-dependent activation of the coaT operator promoter. A CoaR mutant (Ser-Asn-Ser) in a carboxyl-terminal Cys-His-Cys motif bound the coaT operator-promoter but did not activate expression in response to cobalt, implicating thiolate and/or imidazole ligands at these residues in an allosteric cobalt binding site. Deletion of 1 or 2 nucleotides from between near consensus, but with aberrant (20 base pairs) spacing, -10 and -35 elements enhanced expression from the coaT operator-promoter but abolished activation by cobalt-CoaR. It is inferred that cobalt effects a transition in CoaR that underwinds the coaT operator-promoter to realign promoter elements. In the absence of cobalt, CoaR represses expression (approximately 50%). CoaR is a fusion of ancestral MerR (mercury-responsive transcriptional activator)- and precorrin isomerase (enzyme of vitamin B(12) biosynthesis)-related sequences. Expression from the coaT operator-promoter was enhanced in a partial mutant of cbiE (encoding an enzyme preceding precorrin isomerase in B(12) biosynthesis), revealing that this pathway "inhibits" coaT expression. Disruption of coaT reduced cobalt tolerance and increased cytoplasmic (57)Co accumulation. coaT mediated restoration of cobalt tolerance has been used as a selectable marker. PMID- 10464324 TI - Activation of mitogen-activated protein kinase by the A(2A)-adenosine receptor via a rap1-dependent and via a p21(ras)-dependent pathway. AB - The A(2A)-adenosine receptor, a prototypical G(s)-coupled receptor, activates mitogen-activated protein (MAP) kinase in a manner independent of cAMP in primary human endothelial cells. In order to delineate signaling pathways that link the receptor to the regulation of MAP kinase, the human A(2A) receptor was heterologously expressed in Chinese hamster ovary (CHO) and HEK293 cells. In both cell lines, A(2A) agonist-mediated cAMP accumulation was accompanied by activation of the small G protein rap1. However, rap1 mediates A(2A) receptor dependent activation of MAP kinase only in CHO cells, the signaling cascade being composed of G(s), adenylyl cyclase, rap1, and the p68 isoform of B-raf. This isoform was absent in HEK293 cells. Contrary to CHO cells, in HEK293 cells activation of MAP kinase by A(2A) agonists was not mimicked by 8-bromo-cAMP, was independent of Galpha(s), and was associated with activation of p21(ras). Accordingly, overexpression of the inactive S17N mutant of p21(ras) and of a dominant negative version of mSos (the exchange factor of p21(ras)) blocked MAP kinase stimulation by the A(2A) receptor in HEK 293 but not in CHO cells. In spite of the close homology between p21(ras) and rap1, the S17N mutant of rap1 was not dominant negative because (i) overexpression of rap1(S17N) failed to inhibit A(2A) receptor-dependent MAP kinase activation, (ii) rap1(S17N) was recovered in the active form with a GST fusion protein comprising the rap1 binding domain of ralGDS after A(2A) receptor activation, and (iii) A(2A) agonists promoted the association of rap1(S17N) with the 68-kDa isoform of B-raf in CHO cells. We conclude that the A(2A) receptor has the capacity two activate MAP kinase via at least two signaling pathways, which depend on two distinct small G proteins, namely p21(ras) and rap1. Our observations also show that the S17N version of rap1 cannot be assumed a priori to act as a dominant negative interfering mutant. PMID- 10464325 TI - Differential effects of cAMP in neurons and astrocytes. Role of B-raf. AB - Mitogen-activated protein kinase (MAPK) activation provides cell type-specific signals important for cellular differentiation, proliferation, and survival. Cyclic AMP (cAMP) has divergent effects on MAPK activity depending on whether signaling is through Ras/Raf-1 or Rap1/B-raf. We found that central nervous system-derived neurons, but not astrocytes, express B-raf. In neurons, cAMP activated MAPK in a Rap1/B-raf-dependent manner, while in astrocytes, cAMP decreased MAPK activity. Inhibition of MAPK in neurons decreased neuronal growth factor-mediated survival, and activation of MAPK by cAMP analogues rescued neurons from death. Furthermore, constitutive expression of B-raf in astrocytoma cells increased MAPK activation, as seen in neurons, and enhanced proliferation. These data provide the first experimental evidence that B-raf is the molecular switch which dominantly permits differential cAMP-dependent regulation of MAPK in neurons versus astrocytes, with important implications for both survival and proliferation. PMID- 10464326 TI - The anion-stimulated ATPase ArsA shows unisite and multisite catalytic activity. AB - ArsA, an anion-stimulated ATPase, consists of two nucleotide binding domains, A1 in the N terminus and A2 in the C terminus of the protein, connected by a linker. The A1 domain contains a high affinity ATP binding site, whereas the A2 domain has low affinity and it requires the allosteric ligand antimonite for binding ATP. ArsA is known to form a UV-activated adduct with [alpha-(32)P]ATP in the linker region. This study shows that on addition of antimonite, much more adduct is formed. Characterization of the nature of the adduct suggests that it is between ArsA and ADP, instead of ATP, indicating that the adduct formation reflects hydrolysis of ATP. The present study also demonstrates that the A1 domain is capable of carrying out unisite catalysis in the absence of antimonite. On addition of antimonite, multisite catalysis involving both A1 and A2 sites occurs, resulting in a 40-fold increase in ATPase activity. Studies with mutant proteins suggest that the A2 site may be second in the sequence of events, so that its role in catalysis is dependent on a functional A1 site. It is also proposed that ArsA goes through an ATP-bound and an ADP-bound conformation, and the linker region, where ADP binds under both unisite and multisite catalytic conditions, may play an important role in the energy transduction process. PMID- 10464327 TI - Distinct roles for STAT1, STAT3, and STAT5 in differentiation gene induction and apoptosis inhibition by interleukin-9. AB - Interleukin-9 (IL-9) activates three distinct STAT proteins: STAT1, STAT3, and STAT5. This process depends on one tyrosine of the IL-9 receptor, which is necessary for proliferation, gene induction, and inhibition of apoptosis induced by glucocorticoids. By introduction of point mutations in amino acids surrounding this tyrosine, we obtained receptors that activated either STAT5 alone or both STAT1 and STAT3, thus providing us with the possibility to study the respective roles of these factors in the biological activities of IL-9. Both mutant receptors were able to prevent apoptosis, but only the mutant that activated STAT1 and STAT3 was able to support induction of granzyme A and L-selectin. In line with these results, constitutively activated STAT5 blocked glucocorticoid induced apoptosis. In Ba/F3 cells, significant proliferation and pim-1 induction were observed with both STAT-restricted mutants, though proliferation was lower than with the wild-type receptor. These results suggest that survival and cell growth are redundantly controlled by multiple STAT factors, whereas differentiation gene induction is more specifically correlated with individual STAT activation by IL-9. PMID- 10464328 TI - Goodpasture disease. Characterization of a single conformational epitope as the target of pathogenic autoantibodies. AB - Goodpasture disease is a prototype autoimmune disease characterized by the formation of autoantibodies against the heterotrimeric basement membrane collagen type IV, which causes a rapidly progressive glomerulonephritis. The pathogenic antibody response is directed to the non-collagenous (NC1) domain of the alpha3 chain of type IV collagen (alpha3(IV)NC1), but not to the homologous region of the alpha1(IV)NC1. To identify the conformation-dependent immunodominant epitope on the alpha3(IV)NC1, a variety of recombinant NC1 domains were constructed by replacing single residues of alpha3(IV) with the corresponding amino acids from the nonreactive alpha1(IV) chain. Replacement mutations were identified that completely destroyed the Goodpasture epitope in the alpha3(IV) chain. Based on the identification of these critical positions, the epitope was finally reconstructed within the frame of the alpha1(IV) chain. The substitution of nine discontinuous positions in the alpha1(IV)NC1 with amino acid residues from the alpha3 chain resulted in a recombinant construct that was recognized by all patients' sera (n = 20) but by none of the sera from healthy controls (n = 10). This provides, for the first time, the molecular characterization of a single immunodominant conformational epitope recognized by pathogenic autoantibodies in a human autoimmune disease, representing the basis for the development of new epitope-specific strategies in the treatment of Goodpasture disease. PMID- 10464329 TI - Stable overexpression of manganese superoxide dismutase in mitochondria identifies hydrogen peroxide as a major oxidant in the AP-1-mediated induction of matrix-degrading metalloprotease-1. AB - Reactive oxygen species (ROS) are important second messengers for the induction of several genes in a variety of physiological and pathological conditions. Here we addressed the question of whether isolated, unbalanced overexpression of the antioxidant enzyme manganese superoxide dismutase (Mn-SOD) may modulate signal transduction cascades, finally leading to connective tissue degradation, a hallmark in carcinogenesis and aging. Therefore, we generated stably Mn-SOD overexpressing fibroblasts with an up to 4. 6-fold increase in Mn-SOD activity. The Mn-SOD-overexpressing cells revealed specific resistance to the superoxide anion (O-(2))-generating agent paraquat, whereas no resistance to UVA-generated oxidative stress was found. Treatment of the Mn-SOD-overexpressing cells with various ROS-generating systems resulted (due to the enhanced dismutation of superoxide anion to hydrogen peroxide) in an up to 9.5-fold increase in matrix degrading metalloprotease-1 (MMP-1) mRNA levels. A similar increase in MMP-1 mRNA was also seen when the intracellular H(2)O(2) concentration was increased by the inhibition of different H(2)O(2)-detoxifying pathways. Furthermore, prooxidant conditions led to a strong induction of c-jun and c-fos mRNA levels resulting in a 4-fold higher transactivation of the transcription factor AP-1 in the Mn-SOD overexpressing cells. Collectively, we have found that enhanced Mn-SOD activity, via an unbalanced H(2)O(2) overproduction and detoxification, induces MMP-1 mRNA levels, and this effect is at least partly mediated by the DNA recognition sequence AP-1. PMID- 10464330 TI - The carboxyl-terminal domain of receptor-associated protein facilitates proper folding and trafficking of the very low density lipoprotein receptor by interaction with the three amino-terminal ligand-binding repeats of the receptor. AB - The 39-kDa receptor-associated protein (RAP) is a specialized antagonist that inhibits all known ligand interactions with receptors that belong to the low density lipoprotein (LDL) receptor gene family. Recent studies have demonstrated a role for RAP as a molecular chaperone for the LDL receptor-related protein during receptor folding and trafficking within the early secretory pathway. In the present study, we investigated a potential role for RAP as a chaperone for the very low density lipoprotein (VLDL) receptor, another member of the LDL receptor gene family. Using intracellular cross-linking techniques, we found that RAP is associated with newly synthesized VLDL receptor. In the absence of RAP co expression, newly synthesized VLDL receptor exhibited slower trafficking along the early secretory pathway, most likely due to misfolding of the receptor. The role of RAP in the folding of the VLDL receptor was further studied using an anchor-free, soluble VLDL receptor. Metabolic pulse-chase labeling experiments showed that while only 3% of the soluble VLDL receptor was folded and secreted in the absence of RAP co-expression, over 50% of the soluble receptor was secreted in the presence of RAP co-expression. The functions of RAP in VLDL receptor folding and trafficking were mediated by its carboxyl-terminal repeat but not by the amino-terminal and central repeats. Using truncated VLDL receptor constructs, we identified the RAP-binding site within the first three ligand-binding repeats of the VLDL receptor. Thus, our present study demonstrates that RAP serves as a folding and trafficking chaperone for the VLDL receptor via interactions of its carboxyl-terminal repeat with the three amino-terminal ligand-binding repeats of the VLDL receptor. PMID- 10464331 TI - Purification and analysis of authentic CLIP-170 and recombinant fragments. AB - We have purified authentic CLIP-170 (cytoplasmic linker protein of 170 kDa) and fragments comprising functional domains of the protein to characterize the structural basis of the function of CLIP-170. Analysis of authentic CLIP-170 and the recombinant fragments by electron microscopy after glycerol spraying/low angle rotary metal shadowing reveals CLIP-170 as a thin, 135-nm-long molecule with two kinks in its central rod domain, which are approximately equally spaced from the two ends of the protein. The central domain consisting of heptad repeats, which is alpha-helical in nature and forms a 2-stranded coiled-coil, mediates dimerization of CLIP-170. The rod domain harbors two kinks, each spaced approximately 37 nm from the corresponding end of the molecule, thus providing mechanical flexibility to the highly elongated molecule. The N-terminal domain of CLIP-170 binds to microtubules in vitro with a stoichiometry of one dimeric head domain per four tubulin heterodimers. Authentic CLIP-170 binds to microtubules with lower stoichiometry, indicating that the rod and tail domains affect microtubule binding of CLIP-170. These results document that CLIP-170 is a highly elongated polar molecule with the microtubule-binding domain and the organelle interacting domains at opposite ends of the homodimer, thus providing a structural basis for the function of CLIP-170 as a microtubule-organelle linker protein. PMID- 10464332 TI - Fish oil feeding decreases mature sterol regulatory element-binding protein 1 (SREBP-1) by down-regulation of SREBP-1c mRNA in mouse liver. A possible mechanism for down-regulation of lipogenic enzyme mRNAs. AB - Dietary fish oil induces hepatic peroxisomal and microsomal fatty acid oxidation by peroxisome proliferator-activator receptor alpha activation, whereas it down regulates lipogenic gene expression by unknown mechanism(s). Because sterol regulatory element-binding proteins (SREBPs) up-regulated lipogenic genes, investigation was made on the effects of fish oil feeding on SREBPs and sterol regulatory element (SRE)-dependent gene expression in C57BL/6J mice. Three forms of SREBPs, SREBP-1a, -1c, and -2, are expressed in liver, and their truncated mature forms activate transcription of sterol-regulated genes. C57BL/6J mice were divided into three groups; the first group was given a high carbohydrate diet, and the other two groups were given a high fat diet (60% of total energy), with the fat in the form of safflower oil or fish oil, for 5 months. Compared with safflower oil feeding, fish oil feeding decreased triglyceride and cholesterol concentrations in liver. There were no differences in amount of SREBP-1 and -2 in both precursor and mature forms between carbohydrate- and safflower oil-fed mice. However, compared with safflower oil feeding, fish oil feeding reduced the amounts of precursor SREBP-1 in membrane fraction by 90% and of mature SREBP-1 in liver nuclei by 57%. Fish oil feeding also reduced precursor SREBP-2 by 65% but did not alter the amount of mature SREBP-2. Compared with safflower oil feeding, fish oil feeding decreased liver SREBP-1c mRNA level by 86% but did not alter SERBP-1a mRNA. Consistent with decrease of mature SREBP-1, compared with safflower oil feeding, fish oil feeding down-regulated the expression of liver SRE-dependent genes, such as low density lipoprotein receptor, 3-hydroxy-3 methylglutaryl-CoA reductase, 3-hydroxy-3-methylglutaryl-CoA synthase, fatty acid synthase, acetyl-CoA carboxylase, and stearoyl-CoA desaturase-1. These data suggested that in liver, fish oil feeding down-regulates the mature form of SREBP 1 by decreasing SREBP-1c mRNA expression, with corresponding decreases of mRNAs of cholesterologenic and lipogenic enzymes. PMID- 10464333 TI - Genetic evidence for the heterodimeric structure of glucosidase II. The effect of disrupting the subunit-encoding genes on glycoprotein folding. AB - It has been proposed that in rat and murine tissues glucosidase II (GII) is formed by two subunits, GIIalpha and GIIbeta, respectively, responsible for the catalytic activity and the retention of the enzyme in the endoplasmic reticulum (ER). To test this proposal we disrupted genes (gls2alpha(+) and gls2beta(+)) encoding GIIalpha and GIIbeta homologs in Schizosaccharomyces pombe. Both mutant cells (gls2alpha and gls2beta) were completely devoid of GII activity in cell free assays. Nevertheless, N-oligosaccharides formed in intact gls2alpha cells were identified as Glc(2)Man(9)GlcNAc(2) and Glc(2)Man(8)GlcNAc(2), whereas gls2beta cells formed, in addition, small amounts of Glc(1)Man(9)GlcNAc(2). It is suggested that this last compound was formed by GIIalpha transiently present in the ER. Monoglucosylated oligosaccharides facilitated glycoprotein folding in S. pombe as mutants, in which formation of monoglucosylated glycoproteins was completely (gls2alpha) or severely (gls2beta and UDP Glc:glycoprotein:glucosyltransferase null) diminished, showed ER accumulation of misfolded glycoproteins when grown in the absence of exogenous stress as revealed by (a) induction of binding protein-encoding mRNA and (b) accumulation of glycoproteins bearing ER-specific oligosaccharides. Moreover, the same as in mammalian cell systems, formation of monoglucosylated oligosaccharides decreased the folding rate and increased the folding efficiency of glycoproteins as pulse chase experiments revealed that carboxypeptidase Y arrived at a higher rate but in decreased amounts to the vacuoles of gls2alpha than to those of wild type cells. PMID- 10464334 TI - Contact-dependent growth inhibition and apoptosis of epidermal growth factor (EGF) receptor-expressing cells by the membrane-anchored form of heparin-binding EGF-like growth factor. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF) transduces mitogenic signals through the EGF receptor (EGFR). There are two forms of HB-EGF, the membrane-anchored form (pro-HB-EGF) and the soluble form (sHB-EGF). We studied the biological activity of pro-HB-EGF by using a model in which pro-HB EGF-expressing effector cells was co-cultured with EGFR-expressing target cells. The DER cell, an EGFR-expressing derivative of the interleukin-3-dependent hematopoietic 32D cell line, grows well in the presence of EGF or sHB-EGF without IL-3. When DER cells were co-cultured on a monolayer of Vero-H cells overexpressing pro-HB-EGF, growth inhibition and subsequent apoptosis were induced in the DER cells even in the presence of excess amounts of EGF or sHB EGF. Such growth inhibition of DER cells was abrogated when specific antagonists for pro-HB-EGF were added in the culture medium or when direct contact of DER cells with Vero-H cells was prevented, indicating that pro-HB-EGF is involved in this inhibitory effect. Pro-HB-EGF-induced apoptosis of DER cells was also observed even in the presence of IL-3. This rules out the possibility of simple competition between soluble EGFR ligands and pro-HB-EGF. Moreover, 32D cells expressing EGFR mutant composed of the extracellular and the transmembrane domain of EGFR and the cytoplasmic domain of erythropoietin receptor did not undergo apoptosis by co-culture with Vero-H cells, indicating that the inhibitory signal induced by pro-HB-EGF-expressing Vero-H cells is mediated to DER cells via EGFR and that the cytoplasmic domain of EGFR is essential for pro-HB-EGF-induced apoptosis. From these results, we concluded that pro-HB-EGF has unique biological activity through cell-cell contact that is distinct from the activity of sHB-EGF. PMID- 10464335 TI - Phospholipase A(2) modification of low density lipoproteins forms small high density particles with increased affinity for proteoglycans and glycosaminoglycans. AB - The presence of a lipoprotein profile with abundance of small, dense low density lipoproteins (LDL), low levels of high density lipoproteins (HDL), and elevated levels of triglyceride-rich very low density lipoproteins is associated with an increased risk for coronary heart disease. The atherogenicity of small, dense LDL is believed to be one of the main reasons for this association. This particle contains less phospholipids (PL) and unesterified cholesterol than large LDL, and the apoB-100 appears to occupy a more extensive area at its surface. Although there are experiments that suggest a metabolic pathway leading to the overproduction of small, dense LDL, no clear molecular model exists to explain its association with atherogenesis. A current hypothesis is that small, dense LDL, because of its higher affinity for proteoglycans, is entrapped in the intima extracellular matrix and is more susceptible to oxidative modifications than large LDL. Here we describe how a specific reduction of approximately 50% of the PL of a normal buoyant LDL by immobilized phospholipase A(2) (PLA(2)) (EC 3.1.1.4) produces smaller and denser particles without inducing significant lipoprotein aggregation (<5%). These smaller LDL particles display a higher tendency to form nonsoluble complexes with proteoglycans and glycosaminoglycans than the parent LDL. Binding parameters of LDL and glycosaminoglycans and proteoglycans produced by human arterial smooth muscle cells were measured at near to physiological conditions. The PLA(2)-modified LDL has about 2 times higher affinity for the sulfated polysaccharides than control LDL. In addition, incubation of human plasma in the presence of PLA(2) generated smaller LDL and HDL particles compared with the control plasma incubated without PLA(2). These in vitro results indicate that the reduction of surface PL characteristic of small, dense LDL subfractions, besides contributing to its small size and density, may enhance its tendency to be retained by proteoglycans. PMID- 10464336 TI - Regulated degradation of yeast ornithine decarboxylase. AB - Ornithine decarboxylase (ODC) declines in cells that accumulate an excess of polyamines, the downstream products of the enzyme. Superfluous production of polyamines is thus prevented. In animal cells, polyamines reduce ODC activity by accelerating its degradation. Similar down-regulation of ODC activity has been observed in the budding yeast Saccharomyces cerevisiae, but induced degradation has not been documented. Here we show using pulse-chase analysis that the loss of enzyme activity is the result of increased degradation of ODC. Polyamines reduce the half-life of the newly synthesized protein from 3 h to approximately 10 min. Degradation of bulk ODC pools is also accelerated by polyamines, but the absolute rate of turnover is slower, with a half-life of 5 h in untreated and 1 h in treated cells. Newly synthesized ODC polypeptide thus undergoes a process of maturation that renders it relatively resistant to both basal and polyamine induced degradation. Proteasome mutants have a blunted or absent regulatory response, implicating both the core protease and the regulatory cap of the proteasome in induced degradation of yeast ODC. PMID- 10464337 TI - Cell cycle regulation of human CDC6 protein. Intracellular localization, interaction with the human mcm complex, and CDC2 kinase-mediated hyperphosphorylation. AB - The binding of mammalian MCM complexes to chromatin is cell cycle-regulated and under CDC2 kinase negative control. Here, we investigated the properties of mammalian CDC6 protein, a candidate regulator of MCM. The levels of CDC6 were relatively constant during the HeLa cell cycle. In asynchronous cells, CDC6 was mainly detected in the nuclei with immunostaining, but some CDC6 was not extractable with nonionic detergent. In contrast to the chromatin-bound MCM, this fraction of CDC6 was resistant to DNase I treatment, suggesting that it binds to the detergent- and nuclease-resistant nuclear structure. In S phase cells, CDC6 became detectable in the cytoplasm with immunostaining; however, the level of the bound CDC6 was unchanged. In G(2)/M phase cells, the level of the bound CDC6 was still maintained, which was hyperphosphorylated by CDC2 kinase. These data suggest that some CDC6 protein is associated with the specific nuclear structure throughout the cell cycle and that major binding sites on chromatin differ between MCM and CDC6. However, co-immunoprecipitation assays with chemical cross linking indicated that a small part of the chromatin-bound MCM is present close to the bound CDC6. PMID- 10464338 TI - Eosinophil peroxidase nitrates protein tyrosyl residues. Implications for oxidative damage by nitrating intermediates in eosinophilic inflammatory disorders. AB - Eosinophil peroxidase (EPO) has been implicated in promoting oxidative tissue injury in conditions ranging from asthma and other allergic inflammatory disorders to cancer and parasitic/helminthic infections. Studies thus far on this unique peroxidase have primarily focused on its unusual substrate preference for bromide (Br(-)) and the pseudohalide thiocyanate (SCN(-)) forming potent hypohalous acids as cytotoxic oxidants. However, the ability of EPO to generate reactive nitrogen species has not yet been reported. We now demonstrate that EPO readily uses nitrite (NO(2)(-)), a major end-product of nitric oxide ((.)NO) metabolism, as substrate to generate a reactive intermediate that nitrates protein tyrosyl residues in high yield. EPO-catalyzed nitration of tyrosine occurred more readily than bromination at neutral pH, plasma levels of halides, and pathophysiologically relevant concentrations of NO(2)(-). Furthermore, EPO was significantly more effective than MPO at promoting tyrosine nitration in the presence of plasma levels of halides. Whereas recent studies suggest that MPO can also promote protein nitration through indirect oxidation of NO(2)(-) with HOCl, we found no evidence that EPO can indirectly mediate protein nitration by a similar reaction between HOBr and NO(2)(-). EPO-dependent nitration of tyrosine was modulated over a physiologically relevant range of SCN(-) concentrations and was accompanied by formation of tyrosyl radical addition products (e.g. o,o' dityrosine, pulcherosine, trityrosine). The potential role of specific antioxidants and nucleophilic scavengers on yields of tyrosine nitration and bromination by EPO are examined. Thus, EPO may contribute to nitrotyrosine formation in inflammatory conditions characterized by recruitment and activation of eosinophils. PMID- 10464339 TI - Amyloid beta-protein fibrillogenesis. Structure and biological activity of protofibrillar intermediates. AB - Alzheimer's disease is characterized by extensive cerebral amyloid deposition. Amyloid deposits associated with damaged neuropil and blood vessels contain abundant fibrils formed by the amyloid beta-protein (Abeta). Fibrils, both in vitro and in vivo, are neurotoxic. For this reason, substantial effort has been expended to develop therapeutic approaches to control Abeta production and amyloidogenesis. Achievement of the latter goal is facilitated by a rigorous mechanistic understanding of the fibrillogenesis process. Recently, we discovered a novel intermediate in the pathway of Abeta fibril formation, the amyloid protofibril (Walsh, D. M., Lomakin, A., Benedek, G. B., Condron, M. M., and Teplow, D. B. (1997) J. Biol. Chem. 272, 22364-22372). We report here results of studies of the assembly, structure, and biological activity of these polymers. We find that protofibrils: 1) are in equilibrium with low molecular weight Abeta (monomeric or dimeric); 2) have a secondary structure characteristic of amyloid fibrils; 3) appear as beaded chains in rotary shadowed preparations examined electron microscopically; 4) give rise to mature amyloid-like fibrils; and 5) affect the normal metabolism of cultured neurons. The implications of these results for the development of therapies for Alzheimer's disease and for our understanding of fibril assembly are discussed. PMID- 10464341 TI - CAG members receive prestigious janssen awards PMID- 10464340 TI - Homer 1b regulates the trafficking of group I metabotropic glutamate receptors. AB - The molecular basis for glutamate receptor trafficking to the plasma membrane is not understood. In the present study, we demonstrate that Homer 1b (H1b), a constitutively expressed splice form of the immediate early gene product Homer (now termed Homer 1a) regulates the trafficking and surface expression of group I metabotropic glutamate receptors. H1b inhibits surface expression of the metabotropic glutamate receptor mGluR5 in heterologous cells, causing mGluR5 to be retained in the endoplasmic reticulum (ER). In contrast, mGluR5 alone or mGluR5 coexpressed with Homer 1a successfully travels through the secretory pathway to the plasma membrane. In addition, point mutations that disrupt mGluR5 binding to H1b eliminate ER retention of mGluR5, demonstrating that H1b affects metabotropic receptor localization via a direct protein-protein interaction. Electron microscopic analysis reveals that the group I metabotropic receptor mGluR1alpha is significantly enriched in the ER of Purkinje cells, suggesting that a similar mechanism may exist in vivo. Because H1b is found in dendritic spines of neurons, local retention of metabotropic receptors within dendritic ER provides a potential mechanism for regulating synapse-specific expression of group I metabotropic glutamate receptors. PMID- 10464342 TI - Chest pain of esophageal origin. PMID- 10464343 TI - Distribution of morphological changes in collagenous colitis PMID- 10464344 TI - Pancreatic stones: treat or ignore? AB - Painful, chronic pancreatitis is of complex etiology, but increasing clinical experience suggests that removal of pancreatic duct stones in many cases significantly improves patients' symptoms. The development and refinement of therapeutic endoscopic retrograde choledochopancreatography have permitted improved access to the pancreatic duct, which makes the development of new techniques of stone fragmentation and fragment removal a much more successful nonsurgical intervention. A major step forward has been the understanding of the safety and efficacy of pancreatic sphincterotomy, which is necessary for the removal of these difficult stones. The recognition that extracorporeal shock wave lithotripsy can be delivered safely with good efficacy has revolutionized the nonsurgical management of pancreatic duct stones. Nevertheless, advanced and sophisticated therapeutic endoscopy is necessary to achieve clearance of the duct, which can generally be accomplished in the majority of selected patients. State-of-the-art treatments are described, and some new approaches using pancreatoscopy and electrohydrolic lithotripsy are discussed. Newly recognized long term complications are reviewed. Finally, it must be recognized that chronic pancreatitis is an ongoing disease that does not have a simple treatment or cure, and frequently represents a process of remissions and relapses requiring interventions and problem solving. PMID- 10464345 TI - Intrahepatic stones: the percutaneous approach. AB - Intrahepatic stones are prevalent in the Far East, whereas they are infrequently seen in Western countries. Hepatolithiasis can cause recurrent attacks of cholangitis, with a risk of liver abscesses, sepsis or hepatic failure. Immediate biliary decompression can usually be achieved by endoscopic or percutaneous transhepatic drainage. Definitive treatment should aim for complete elimination of bile stasis and removal of all stones. Hepatic resection promises the best long term results when the disease is limited to segments or the left liver lobe. Endoscopic retrograde choledochopancreatography is not well established for intrahepatic stones because of frequent failures due to associated biliary strictures, angulated ducts or peripherally impacted concrements. In contrast, percutaneous procedures can be easily performed through a T tube tract for residual stones after surgery. Establishment of a transhepatic fistula allows a targeted approach to liver segments with catheters or miniscopes, without the need for laparotomy. Biliary strictures can be dilated with balloons, and intrahepatic stones can be removed with baskets under fluoroscopic or cholangioscopic control. These techniques can be combined with electrohydraulic lithotripsy or laser lithotripsy for disintegration of impacted calculi. The risk of stone recurrence is particularly high in patients with associated biliary stenoses. Temporary or long term transhepatic intubation is a promising approach in these cases. The optimal management of intrahepatic stones remains a challenging task that requires an experienced team of gastroenterologists, surgeons and radiologists. PMID- 10464346 TI - Colonoscopy 'my way': preparation, anticoagulants, antibiotics and sedation. AB - Colonoscopy was introduced in the 1960s. The facility with which this technique is performed has been enhanced by vast improvements in instrumentation. In spite of this, physician attitudes concerning colonoscopy have changed little over the past several decades. The diet for precolonoscopic preparation has not been altered for 30 years. Colonoscopists have a great reluctance to use a new preparation instead of the 4 L electrolyte solution, perhaps because this was such a significant advance in colonoscopic cleansing, its predecessor being castor oil and enemas. Physicians continue to be wary of the patient who is taking acetylsalicylic acid in the absence of any studies that show that this is detrimental for polypectomy. The management of the patient on warfarin anticoagulation remains a subject for debate. As for antibiotic prophylaxis, most endoscopy units do not have a standardized approach, although there are good guidelines that, if followed, should decrease the risk of infective endocarditis. Sedation for the endoscopic examination is usually administered by the colonoscopist, although anesthesiologists may, in some countries (and in some defined areas of the United States) be the primary administrators of sedation and analgesia. The present article is a personal approach to the following issues: the preparation of the colon for an examination, current thoughts about anticoagulation and acetylsalicylic acid, antibiotic prophylaxis for colonoscopy and the technique for sedation out of the hospital. PMID- 10464347 TI - Endoscopic mucosal resection using a cap: techniques for use and preventing perforation. AB - Endoscopic mucosal resection (EMR) is one of several local treatments that provide a specimen for histopathological analysis. The authors developed a technique of EMR using a transparent plastic cap (EMRC) in 1992. By using the EMRC procedure, any part of the gastrointestinal tract mucosa can be easily accessed. The technical details of EMRC are described. The authors have performed EMR in 380 cases of gastrointestinal lesions. The most serious complication may be perforation. Two perforations (one in the esophagus and one in the colon) have occurred. By evaluating recorded videotapes, it was determined that the lack of submucosal saline injection was the major cause. Therefore, large volume injection, which creates a large bleb and potentially reduces the risk of perforation, is recommended. Furthermore, target mucosa should be strangulated at the middle part of the created bleb (never strangulated at the base). Particularly in the colon, injecting a sufficient volume of saline and controlling the power of suction are extremely important, because the cap on the colonoscope is relatively large in size. PMID- 10464348 TI - Endoscopic pancreatic stenting in pancreatic cancer. AB - Most pancreatic carcinomas are unresectable at the time of diagnosis; therefore, palliative treatment is very often the main concern of clinicians in this setting. The main symptoms resulting in the need for palliation in pancreatic cancer are obstructive jaundice, duodenal obstruction and pain. Therapeutic endoscopy plays a major role in the palliation of obstructive jaundice by stent placement into the biliary ducts. Initial experience has also been gained recently with endoscopic placement of expandable metallic stents to treat gastric outlet obstruction. Much less is known about the possible role of endoscopic pancreatic stenting in patients with unresectable pancreatic carcinoma. The main indication for pancreatic ductal stenting is 'obstructive' pain related to meals in patients with dilated main pancreatic duct beyond the stricture and intraluminal brachyradiotherapy. The technique of endoscopic pancreatic stenting does not substantially differ from that applied on the biliary tree. When technically possible, placement of 10 French plastic stents is preferred. According to the authors' indications, only about 15% of patients with advanced pancreatic cancer (55 of 355 in the present study) may potentially benefit from this technique. Pancreatic stenting may be obtained in more than 80% of these selected patients, with low morbidity (less than 10%) and no procedure-related mortality. According to the authors of the present and other studies reported in the literature, about 60% of patients treated because of 'obstructive' pain become symptom-free, and another 20% to 25% significantly reduce the amount of analgesic drugs required. Intraluminal brachyradiotherapy with 192iridium in the main pancreatic duct is a feasible and safe method to deliver high radiation doses to the tumour while sparing adjacent organs. Brachyradiotherapy may be performed alone or in conjunction with external beam radiotherapy. Because of the small number of patients suitable for this treatment, only a multicentre study will be able to detect whether intraluminal brachyradiotherapy in pancreatic cancer may have any positive impact on survival. PMID- 10464349 TI - Survey of current practices among members of CAG in the follow-up of patients diagnosed with gastric ulcer. AB - INTRODUCTION: Guidelines recommend a policy of endoscopic follow-up of all gastric ulcers until healing. Analysis of data from the Alberta Endoscopy Project indicates that fewer than 50% of patients diagnosed with benign gastric ulcer had undergone a repeat procedure. The practice and attitudes of physician members of the Canadian Association of Gastroenterology (CAG) on the follow-up of such patients were assessed. METHODS: A self-administered questionnaire was mailed to members of CAG. Respondents were asked to indicate their practice setting and to estimate the proportion of gastric ulcer patients in whom they perform follow-up endoscopy. They were also asked to indicate factors influencing this choice, including the role of Helicobacter pylori. RESULTS: Fifty-seven per cent of 220 respondents indicated that they perform repeat endoscopy in 95% to 100% of individuals with benign gastric ulcer. The most common reasons influencing this choice were to ensure healing (86.3%) and to confirm the benign nature of the lesion (79.5%). Nonsteroidal anti-inflammatory drug (NSAID) use (83.2%) and patient ill health (62.9%) were the most common reasons for not repeating the endoscopy. Twenty per cent of individuals indicated that H pylori had influenced a change in their practice. DISCUSSION: Physicians vary widely in their follow-up of benign gastric ulcer. Studies on the occurrence of gastric cancer in this setting are not unanimous in their conclusions. Subgroups of patients with NSAID exposure and successfully eradicated H pylori infection may have a lower risk of malignancy. Studies to confirm this are warranted, and modified guidelines may be appropriate. PMID- 10464350 TI - East meets West: what is early cancer? AB - It has become increasingly apparent that the Japanese and Western systems of classifying dysplasia and carcinoma in the gastrointestinal tract are not the same. The implication of these differences is that in an article in a Western journal on gastrointestinal 'cancer' originating from Japan, it is often impossible to repeat the study to confirm or refute it because of these differences in definitions. 'Carcinoma' is diagnosed in Japan by virtue of its structural and cytological features but by invasion in the Western system. Adenoma does not mean a dysplastic lump in the Japanese system (although it can), but in most cases is similar to low grade dysplasia irrespective of the macroscopic and/or endoscopic appearances (hence flat and depressed adenomas in the Japanese system); however, most examples of high grade dysplasia in the Western system, as well as some low grade dysplasia, are 'cancer' in the Japanese system. Conversely, both have conceptual areas that are useful in the other's system. PMID- 10464351 TI - The ins and outs of therapeutic endoscopy: from the 11th International Course on Therapeutic Endoscopy, October 14 to 17, 1998. PMID- 10464352 TI - Intestinal inflammation and the gut microflora. AB - The idea that the enteric microflora play a role in the pathogenesis or pathophysiology of inflammatory bowel disease (IBD) is not new. Indeed, identification of an infective cause for chronic IBD, and particularly for Crohn's disease, has been the focus of extensive research efforts. During the 1990s, there has been a noticeable re-emergence of interest in the link between bacteria and functional bowel disorders, and the value of antibiotic therapy to treat gut inflammatory disorders. A variety of experimental evidence from both laboratory model systems and clinical investigations is reviewed with respect to a pivotal role for enteric bacteria in gut inflammation. The voluminous scientific literature on this subject precludes any comprehensive synopsis of the area; instead, pertinent studies are cited to illustrate the ability of bacteria and their products to evoke or exacerbate gut inflammation. PMID- 10464353 TI - Fun facts: gems and pearls from the Annual Postgraduate Course of the American College of Gastroenterology: Boston, October 10 to 11, 1998. PMID- 10464354 TI - Common mistakes of affluent investors and how to avoid them. PMID- 10464355 TI - Chronic electrical stimulation by a cochlear implant promotes survival of spiral ganglion neurons after neonatal deafness. AB - This investigation examined the consequences of neonatal deafness and chronic intracochlear electrical stimulation delivered by a cochlear implant during maturation. Kittens were bilaterally deafened by an ototoxic drug administered daily for 2 weeks immediately after birth. Unilateral electrical stimulation was initiated at 7-10 weeks of age and continued over periods of 22-47 weeks (4 hours/day; 5 days/week). Bipolar intracochlear electrodes delivered one of several different electrical signals designed to be temporally challenging to the central auditory system. Morphometric evaluation of spiral ganglion (SG) cell somata within Rosenthal's canal demonstrated a mean of approximately 50% of normal cell density maintained in the chronically stimulated ears, compared with approximately 30% on the control deafened side. This 20% difference in density was highly significant and was greater than differences reported in earlier studies using 30 pps stimulation delivered by either intracochlear bipolar or round window monopolar electrodes. However, the duration of stimulation was also longer in the present study, so it is unclear to what extent the nature of the temporally challenging stimulation vs. its duration contributed to the marked increase in survival. Measurements of the SG cell somata revealed a pronounced decrease in cell diameter in neonatally deafened cats studied about 1 year after deafening, and an additional decrease after long-term deafness (2.5-6.5 years). Furthermore, in the cochlear regions with the greatest stimulation-induced differences in SG cell density, direct measurements of cross-sectional soma area of the largest cells revealed that cells were significantly larger in the stimulated ears. Thus, in addition to the marked increase in the number of surviving SG cells, larger soma area contributed modestly to the pronounced increase in neural density following chronic electrical stimulation. PMID- 10464356 TI - Opioid receptor-like (ORL1) receptor distribution in the rat central nervous system: comparison of ORL1 receptor mRNA expression with (125)I-[(14)Tyr] orphanin FQ binding. AB - The recently discovered neuropeptide orphanin FQ (OFQ), and its opioid receptor like (ORL1) receptor, exhibit structural features suggestive of the micro, kappa, and delta opioid systems. The anatomic distribution of OFQ immunoreactivity and mRNA expression has been reported recently. In the present analysis, we compare the distribution of orphanin receptor mRNA expression with that of orphanin FQ binding at the ORL1 receptor in the adult rat central nervous system (CNS). By using in vitro receptor autoradiography with (125)I-[(14)Tyr]-OFQ as the radioligand, orphanin receptor binding was analyzed throughout the rat CNS. Orphanin binding sites were densest in several cortical regions, the anterior olfactory nucleus, lateral septum, ventral forebrain, several hypothalamic nuclei, hippocampal formation, basolateral and medial amygdala, central gray, pontine nuclei, interpeduncular nucleus, substantia nigra, raphe complex, locus coeruleus, vestibular nuclear complex, and the spinal cord. By using in situ hybridization, cells expressing ORL1 mRNA were most numerous throughout multiple cortical regions, the anterior olfactory nucleus, lateral septum, endopiriform nucleus, ventral forebrain, multiple hypothalamic nuclei, nucleus of the lateral olfactory tract, medial amygdala, hippocampal formation, substantia nigra, ventral tegmental area, central gray, raphe complex, locus coeruleus, multiple brainstem motor nuclei, inferior olive, deep cerebellar nuclei, vestibular nuclear complex, nucleus of the solitary tract, reticular formation, dorsal root ganglia, and spinal cord. The diffuse distribution of ORL1 mRNA and binding supports an extensive role for orphanin FQ in a multitude of CNS functions, including motor and balance control, reinforcement and reward, nociception, the stress response, sexual behavior, aggression, and autonomic control of physiologic processes. PMID- 10464357 TI - Chemically distinct preganglionic inputs to iris-projecting postganglionic neurons in the rat: A light and electron microscopic study. AB - Individual autonomic postganglionic neurons are surrounded by pericellular baskets of preganglionic terminals that are easily identifiable with the light microscope. It has been assumed that the target cell of a pericellular basket of preganglionic terminals is the neuron at the centre of the basket. This assumption has enabled the connectivity of preganglionic neurons to be determined at the light microscopic level. However, if the preganglionic terminals in a pericellular basket make synapses with the dendrites of nearby, but functionally different, postganglionic neurons, then the conclusions of light microscopic studies are far less certain. We have used a serial section ultrastructural study to determine the target of the preganglionic pericellular basket in a situation where the apparent target cell is surrounded by neurons of dissimilar function. In the rat superior cervical ganglion, postganglionic neurons projecting to the iris were identified, using retrograde tracers, as single neurons (i.e., not in clusters). We have used immunohistochemistry to show that iris-projecting neurons are surrounded by preganglionic nerve terminals containing calcitonin gene related peptide (CGRP). We have demonstrated that the pericellular basket of CGRP immunoreactive preganglionic terminals provides inputs only to the soma at the centre of the basket and not to the dendrites of surrounding neurons. This suggests that, in autonomic ganglia, light microscopic identification of the preganglionic terminal baskets is likely to be a reliable method for identifying the targets of subclasses of preganglionic neurons. PMID- 10464358 TI - Retinal axon regeneration in peripheral nerve, tectal, and muscle grafts in adult rats. AB - This study examined whether prior regenerative growth through peripheral nerve (PN) bridging grafts influenced the specificity with which lesioned adult rat retinal ganglion cell (RGC) axons grew into co-grafts of developing target tissue (fetal superior colliculus). Growth into nontarget (muscle) tissue was also examined. Autologous PN was grafted onto the transected optic nerve. After 14 days, the distal ends of the PNs were placed next to, or inserted into, embryonic tectal tissue or into autologous muscle grafts placed in frontal cortex cavities. Host retinal projections were examined 3-8 months later using anterograde and retrograde tracing techniques. In rats in which there was good apposition between PN and tectal tissue, small numbers of RGC axons were observed growing into the tectal grafts (maximum distance of 180 microm). No evidence of specific innervation of appropriate target regions within tectal grafts was detected, even though such regions (identified by acetylcholinesterase histochemistry) were often located close to the PN grafts. In rats with PN/muscle co-grafts, the extent of retinal axon outgrowth was greater (up to 465 microm from the PN tip) and labelled profiles that resembled motor endplates were seen contacting muscle fibres. Previous studies have shown that spontaneously regenerating RGC axons consistently and selectively innervate appropriate target areas in fetal tectal tissue grafted directly into optic tract lesion cavities. Together, the data suggest that exposure to a PN environment may have reduced the extent of adult retinal axon growth into fetal tectal transplants and affected the way regenerating axons responded to specific developmental cues expressed by target cells in the co-grafted tissue. PMID- 10464359 TI - Projections from the upper lumbar cord to the cerebellar nuclei in the rat, studied by anterograde axonal tracing. AB - The spinocerebellar tracts arising from the upper lumbar cord consist of the dorsal and the ventral spinocerebellar tracts (DSCT and VSCT), which ascend ipsilaterally and contralaterally, respectively. By using anterograde labeling with biotinylated dextran in the rat, this study examined whether the lumbar DSCT and the VSCT project to the cerebellar nuclei. Injections of the tracer were made unilaterally at levels between the L1 and L3 segments, with diffusion to either a rostral or a caudal segment. The injections resulted in bilateral labeling of axon terminals in the cerebellar nuclei. In the medial nucleus, labeled terminals were distributed in medial, ventral, and ventrolateral parts of the middle subdivision and ventral parts of the caudomedial subdivision. In the anterior interpositus nucleus, they were distributed in medial and dorsomedial parts throughout the rostrocaudal extent. Labeled terminals were seen within the dorsomedial crest region. In the posterior interpositus nucleus, labeled terminals were seen in the rostromedial extension, the caudomedial part, and the caudal pole. Labeled terminals were seen in the hilus and the ventral part of the lateral nucleus. Projections of the DSCT and the VSCT to these regions were confirmed after tracer injections preceded by sectioning of either tract. Both tracts projected bilaterally, but the DSCT projected mainly ipsilaterally. The present study suggests that the spinocerebellar tracts originating from the upper lumbar cord (the lumbar DSCT and the VSCT) project to specific areas of the cerebellar nuclei to transmit information about the peripheral and central events during the movement of hindlimbs. PMID- 10464360 TI - Ultrastructural localization of beta-arrestin-1 and -2 in rat lumbar spinal cord. AB - beta-arrestins play significant roles in agonist-mediated desensitization of G protein-coupled receptors. Although the presence of beta-arrestin subtypes, beta arrestin-1 and(- 2) in rat brain has been studied extensively, their existence in the spinal cord has not been described. In the current study, we performed immunohistochemical analyses of beta-arrestins at both light and electron microscopic levels using rat lumbar 1-2 spinal cord segments. Intense immunoreactivity for beta-arrestin-1 was found in the motoneurons in lamina IX of the ventral horn and elongated cells in the dorsal nucleus of Clarke. Modest immunoreactivity was detected among the neurons of laminae V and VII/VIII, and weaker immunoreactivity in laminae III, IV, and X. beta-arrestin-2 immunoreactivity was also distributed through laminae III-X in the order of IX > dorsal nucleus of Clarke > V > VII/VIII > IV > III > X. Laminae I and II did not show immunoreactivity. At the electron microscopic level, both beta-arrestin immunoreactive and nonimmunoreactive dendrites were observed, whereas axons and terminal boutons were devoid of immunoreactivity. In immunoreactive dendrites most beta-arrestin immunoreactivity was distributed throughout the cytoplasm, demonstrating their association with microtubules. In addition, strong immunoreactivity was often found at postsynaptic densities. Our results thus suggest beta-arrestins' possible involvement in both motor and sensory mechanisms at the postsynaptic level in rat lumbar spinal cord. PMID- 10464361 TI - Altered development of dopaminergic cells in the retina of weaver mice. AB - Postnatal degeneration of dopaminergic (DA) cells is known to occur in mesencephalic nuclei of mutant weaver mice, whereas retinal DA content is reported to be unchanged in the adult animal. To determine whether morphological changes occur in the weaver retinal DA system, we compared weaver and control developing and adult retinas after tyrosine hydroxylase (TH) immunohistochemistry. The density and distribution of DA cells were analyzed using Dirichlet tessellation. Not only was no DA cell loss found in adult weaver retinas, but we even observed an increase in DA cells in weaver compared to control retinas between postnatal days 14 and 30. Furthermore, some unusual features were found during the latter period: atypical cells (representing a maximum of 12% of the whole DA cell population) were observed, and these differed from typical DA cells in terms of both location (slightly more external within the inner nuclear layer) and appearance (flat somata, round and clear nuclei, thick dendritic trunks emerging laterally and giving rise to horizontal processes). Some of the atypical cells were intermingled in a delicate network lying in a more outer focal plane than the main DA plexus. The expression of GIRK2, a G protein-related inward rectifying K(+) channel responsible for the weaver syndrome, was investigated. Although no GIRK2 labeling was demonstrated in DA cells, its possible involvement in the transient disturbances observed in the weaver DA retinal system is discussed. PMID- 10464362 TI - Neuronal control of the gastric sling muscle of the guinea pig. AB - The gastric sling (oblique) muscle (GSM), located close to the lower esophageal sphincter (LES), is involved in gastric motor function and may cooperate with the LES in controlling propulsion between the esophagus and stomach. Neuronal pathways and transmission to the GSM were investigated in isolated esophagus stomach preparations by using intracellular recording with the focal electrical stimulation and neuroanatomical tracing method. Focal stimulation on the GSM evoked inhibitory junction potentials (IJPs) that were reduced to 45% by 100 microM N-nitro-L-arginine and subsequently blocked by 0.5 microM apamin, thereby unmasking excitatory junction potentials (EJPs), which were abolished by 1 microM hyoscine. Vagal and esophageal stimulation evoked IJPs that were blocked by 100 microM hexamethonium. Vagal stimulation also evoked EJPs after blockade of IJPs. Application of 1,1'-didodecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate to the GSM labeled muscle motor neurons located in the stomach mainly close to the GSM, with a few neurons (2%) in the esophagus. The majority (79%) of labeled neurons were immunoreactive for choline acetyltransferase and, hence, excitatory motor neurons. Inhibitory motor neurons (nitric oxide synthase immunoreactive; 15%) were clustered in the midline near the gastroesophageal region. These results demonstrate that the GSM is innervated primarily by gastric excitatory and inhibitory motor neurons and some esophageal neurons. Both excitatory (acetylcholine) and inhibitory (nitric oxide and apamin-sensitive component) transmission can be activated via vagal-enteric pathways. PMID- 10464363 TI - Overexpression of Bcl-2 is neuroprotective after experimental brain injury in transgenic mice. AB - The cell death regulatory protein, Bcl-2, has been suggested to participate in the pathophysiology of various neurological disorders, including traumatic brain injury (TBI). The cognitive function and histopathologic sequelae after controlled cortical impact brain injury were evaluated in transgenic (TG) mice that overexpress human Bcl-2 protein (n = 13) and their wild type (WT) controls (n = 9). Although brain-injured Bcl-2 TG mice exhibited similar posttraumatic deficits in a Morris water maze (MWM) test of spatial memory as their WT counterparts at 1 week postinjury, the preinjury learning ability of Bcl-2 TG mice was impaired significantly compared with their WT littermates (P < 0.05). In contrast, histopathologic analysis revealed significantly attenuated tissue loss in the ipsilateral hemisphere (p < 0.01) and decreased tissue loss in ipsilateral hippocampal area CA3 (P < 0.001) and the dentate gyrus (P < 0.01) in brain injured Bcl-2 TG mice compared with brain-injured WT mice. Immunohistochemical evaluation of glial fibrillary acidic protein also revealed a significant decrease in reactive astrocytosis in the ipsilateral dorsal thalamus (P < 0.05) and the ventral thalamus (P < 0.01) in brain-injured Bcl-2 TG mice. These results suggest that overexpression of Bcl-2 protein may play a protective role in neuropathologic sequelae after TBI. PMID- 10464365 TI - Erratum AB - Hahm J-O, Cramer KS, Sur M. 1999. Pattern formation by retinal afferents in the ferret lateral geniculate nucleus: developmental segregation and the role of N methyl-D-aspartate receptors. J Comp Neurol 411:327-345. Due to a printer's error, Figures 6 and 12 were transposed after the proofs of the article had been approved by the author. The correct figures and legends are reprinted on the following pages. The publisher regrets the error. PMID- 10464364 TI - Neurokinin-1 receptor localisation in guinea pig autonomic ganglia. AB - We have used multiple-labelling immunohistochemistry and confocal microscopy to determine the distribution of immunoreactivity to the tachykinin neurokinin-1 (NK(1)) receptors in guinea pig sympathetic ganglia. Although nerve fibres containing immunoreactivity to substance P were common in all ganglia except the superior cervical ganglia, most neurons expressing NK(1) receptor immunoreactivity were not closely surrounded by pericellular baskets of substance P-immunoreactive boutons. Conversely, many neurons surrounded by baskets of substance P-immunoreactive boutons lacked NK(1) immunoreactivity. In the coeliac and inferior mesenteric ganglia, NK(1) receptor expression was restricted almost entirely to noradrenergic neurons that contained somatostatin immunoreactivity and projected to the enteric plexuses. In the lumbar chain and paracervical ganglia, NK(1) immunoreactivity was expressed by nonnoradrenergic vasodilator neurons containing immunoreactivity to vasoactive intestinal peptide. Taken together, our results show that sympathetic neurons in different functional pathways express NK(1) receptor immunoreactivity. However, the neurons that could respond to endogenously released substance P through NK(1) receptors may be distant from presynaptic release sites. These observations suggest that, in sympathetic ganglia, substance P may modulate ganglionic transmission through heterosynaptic actions on NK(1) receptors. PMID- 10464366 TI - Corrigendum AB - In a previous manuscript the staining pattern of a monoclonal antibody generated to the N-terminal sequence of the ORL-1 receptor was described in the rat CNS (Anton et al., 1996). Subsequently, this antibody was used in double-label confocal experiments to map mu opioid and ORL-1 receptors in areas of brain associated with pain processing (Monteillet-Agius et al., 1998). In order to stain sections in mice, the monoclonal antibody was biotinylated, and as one of the specificity controls, brain sections from mutant mice lacking the ORL-1 receptor were stained. Mice null for both exon 2 and 3 of the ORL-1 receptor (generated by Zhenping Chen in John Pintar's laboratory, UMDNJ-Robert Wood Johnson Medical School, unpublished) as well as exon 1 knockout mice, which eliminates the epitope to which the antibody was raised, (Nishi et al., 1997) were tested and the same results obtained. The biotinylated antibody showed a staining pattern in the ORL-1 knockout mice indistinguishable from wild-type mice and similar to the pattern found in rat brain. Although the CNS staining pattern in rat was consistent with the distribution of mRNA for the receptor, the retained staining pattern in ORL-1 null mice demonstrates that the monoclonal antibody is recognizing a protein distinct from the ORL-1 receptor. PMID- 10464368 TI - Embryonic development of connections in turtle pallium. AB - We are interested in similarities and conserved mechanisms in early development of the reptilian and mammalian thalamocortical connections. We set out to analyse connectivity in embryonic turtle brains (Pseudemys scripta elegans, between stages 17 and 25), by using carbocyanine dye tracing. From the earliest stages studied, labelling from dorsal and ventral thalamus revealed backlabelled cells among developing thalamic fibres within the lateral forebrain bundle and striatum, which had similar morphology to backlabelled internal capsule cells in embryonic rat (Molnar and Cordery, 1999). However, thalamic crystal placements did not label cells in the dorsal ventricular ridge (DVR) at any stage examined. Crystal placements into both dorsal and lateral cortex labelled cells in the DVR and, reciprocally, DVR crystal placements labelled cells in the dorsal and lateral cortices. Retrograde labelling revealed that thalamic fibres arrive in the DVR and dorsal cortex by stage 19. The DVR received projections from the nucleus rotundus and the dorsal cortex exclusively from the perirotundal complex (including lateral geniculate nucleus). Thalamic fibres show this remarkable degree of specificity from the earliest stage we could examine with selective retrograde labelling (stage 19). Our study demonstrates that axons of similar cells are among the first to reach dorsal and ventral thalamus in mammals and reptiles. Our connectional analysis in turtle suggests that some cells of the mammalian primitive internal capsule are homologous to a cell group within the reptilian lateral forebrain bundle and striatum and that diverse vertebrate brains might use a highly conserved pattern of early thalamocortical development. PMID- 10464367 TI - Connections between cells of the internal capsule, thalamus, and cerebral cortex in embryonic rat. AB - The aim of our study is to understand the development of the earliest connections in the mammalian pallium by documenting the distribution of cells and fibres labelled from the dorsal and ventral thalamus, internal capsule, perirhinal, and dorsal cortex during the period between embryonic day (E) 14 and 17 by using carbocyanine dye tracing in fixed embryonic rat brains. Dye placed in the thalamus of E14 brains backlabels cells in the thalamic reticular nucleus and within the primitive internal capsule. Both anterograde and retrograde tracing confirmed that the first corticofugal projections reach the internal capsule by E14. At E15-E16, after the first cortical plate cells have migrated into the lateral cortex, some cells of the cortical plate and subplate and marginal zone, are backlabelled from the internal capsule, but still not from the dorsal thalamus, even with very long incubation periods. Crystal placement into the perirhinal cortex at E14-E15 labels numerous cells within the internal capsule, whereas no such cells are revealed from dorsal cerebral cortex until E17, suggesting that internal capsule cells establish early connections with the perirhinal and ventral but not dorsal cortex. We propose that the growth of axons from cortex to dorsal thalamus is delayed in two regions: first from E14-E15 at the lateral entrance of the internal capsule and then, from E16, closer to the thalamus, probably within the thalamic reticular nucleus. Subplate projections reach the proximity of the diencephalon at an early stage, but they might never enter the dorsal thalamus. PMID- 10464369 TI - Projections from the superior colliculus motor map to omnipause neurons in monkey. AB - Descending projections from the superior colliculus (SC) motor map to the saccadic omnipause neurons (OPNs) were examined in monkeys by using anterograde transport of tritiated leucine. The SC was divided into three zones: the rostral pole of the motor map, a small horizontal saccade zone in central SC, and a large horizontal saccade zone in caudal SC. Tracer injections into the intermediate layers of the three zones led to different patterns of silver grain deposits in and around nucleus raphe interpositus (RIP), which contains the OPNs: 1) From the rostral pole of the motor map, coarse axon branches of the crossed predorsal bundle spread medially into the RIP, branched, and terminated predominantly unilaterally over cells on the same side. 2) From the small horizontal saccade zone, the axon branches were of a finer caliber and terminated diffusely in the RIP, mainly on the same side. 3) From the large horizontal saccade zone, no terminal labeling was found within the RIP. 4) From the rostral pole of the motor map and small horizontal saccade zone, fiber branches from the ipsilateral descending pathway terminated diffusely over RIP. 5) In addition, terminal labeling in reticulospinal areas of the pons and medulla increased in parallel with the size of the saccade according to the SC motor map. The results suggest that there are multiple projections directly onto OPNs from the rostral SC but not from the caudal SC associated with large gaze shifts. The efferents from the rostral pole of the motor map may subserve the suppression of saccades during visual fixation, and those from the small horizontal saccade zone could inhibit anatagonist premotor circuits. PMID- 10464370 TI - Divergent axon collaterals from the rostral superior colliculus to the pretectal accommodation-related areas and the omnipause neuron area in the cat. AB - Results of previous studies have suggested that the rostral superior colliculus (SC), which corresponds to the representation of the central visual field, is involved in the control of accommodation and active fixation. To clarify the functional interaction between accommodation and active fixation in the rostral SC, this study was performed to investigate the axon collaterals diverging from the rostral SC to the pretectal accommodation-related areas and the omnipause neuron area in the raphe interpositus (RIP) of the cat by using a fluorescent double-labeling technique. This study was conducted on four cats, weighing 2.5 3.5 kg. Retrogradely labeled neurons in the SC were examined following injections of Fast Blue into the RIP and Diamidino Yellow into the pretectal accommodation related areas, which were identified with microstimulation techniques. Neurons projecting to the RIP were located mainly in the intermediate layers in the rostral SC, whereas neurons projecting to the pretectal accommodation-related areas were scattered in the superficial and intermediate layers of the rostral SC. The population of double-labeled neurons was highest in the rostral SC, which corresponds to the representation of the central visual field. The presence of double-labeled cells indicated that the accommodation-related area in the rostral SC contains neurons whose axons collateralize to project to both the pretectal accommodation-related areas and the ominipause neuron area in the RIP. Neurons in the rostral SC may be involved in the interaction between accommodation and active fixation. PMID- 10464371 TI - Gaze shifts in three-dimensional space: A closer look at the superior colliculus. PMID- 10464372 TI - Two types of interneurons in the cat visual thalamus are distinguished by morphology, synaptic connections, and nitric oxide synthase content. AB - The distribution of the neuronal form of the nitric oxide-synthesizing enzyme, brain nitric oxide synthase (BNOS), was examined in the cat thalamus by using immunocytochemical techniques. BNOS was found in both cells and fibers throughout the visual thalamus. BNOS-stained cells were found consistently in the C laminae of the lateral geniculate nucleus (LGN), the pulvinar nucleus, and the lateral posterior nucleus (LP). In the A laminae of the LGN, variable numbers of BNOS stained cells also could be detected. BNOS-stained cells were identified as a subset of interneurons because they all stained for glutamic acid decarboxylase (GAD), but not all GAD-stained cells contained BNOS. The average soma area of BNOS-stained cells was slightly greater than the average soma area of GAD-stained cells. BNOS-stained cells display a distinctive dendritic morphology, which is consistent with previous descriptions of class V neurons (Updyke [1979] J. Comp. Neurol. 186:603-619); they have widespread but fairly sparse arbors of thin, somewhat beaded dendrites. BNOS-stained cells participate in a distinct synaptic circuitry. Although many GAD-stained profiles are filled with vesicles and participate in complex synaptic arrangements, known as glomeruli, BNOS-stained dendrites contain small clusters of vesicles and form dendrodendritic contacts in the extraglomerular neuropil. Thus, there appear to be at least two types of gamma-aminobutyric acidergic interneurons in the visual thalamus of the cat. Interneurons that do not contain BNOS (class III morphology) may exert their effects primarily within synaptic glomeruli (Hamos et al. [1985] Nature 317:618 621), whereas interneurons that contain BNOS (class V morphology) contribute primarily to the extraglomerular neuropil. PMID- 10464373 TI - Cu/Zn SOD deficiency potentiates hearing loss and cochlear pathology in aged 129,CD-1 mice. AB - Copper/zinc superoxide dismutase (Cu/Zn SOD) is a first-line defense against free radical damage in the cochlea and other tissues. To determine whether deficiencies in Cu/Zn SOD increase age-related hearing loss and cochlear pathology, we collected auditory brainstem responses (ABRs) and determined cochlear hair cell loss in 13-month-old 129/CD-1 mice with (a) no measurable Cu/Zn SOD activity (homozygous knockout mice), (b) 50% reduction of Cu/Zn SOD (heterozygous knockout mice), and (c) normal levels of Cu/Zn SOD (wild-type mice). ABRs were obtained by using 4-, 8-, 16-, and 32-kHz tone bursts. Cochleas were harvested immediately after testing, and separate counts were made of inner and outer hair cells. Compared with wild-type mice, homozygous and heterozygous knockout mice exhibited significant threshold elevations and greater hair cell loss. Phenotypic variability was higher among heterozygous knockout mice than among wild-type or homozygous knockout mice. Separate groups of wild-type and homozygous knockout mice were examined for loss of spiral ganglion cells and eighth nerve fibers. At 13 months of age, both wild-type and knockout mice had significantly fewer nerve fibers than did 2-month-old wild-type mice, with significantly greater loss in aged knockout mice than in aged wild-type mice. Thirteen-month-old knockout mice also had a significant loss of spiral ganglion cells compared with 2-month-old wild-type mice. The results indicate that Cu/Zn SOD deficiencies increase the vulnerability of the cochlea to damage associated with normal aging, presumably through metabolic pathways involving the superoxide radical. PMID- 10464374 TI - Distinct neurochemical populations in the rat central nucleus of the amygdala and bed nucleus of the stria terminalis: evidence for their selective activation by interleukin-1beta. AB - The lateral division of the central nucleus of the amygdala (CEAl) and the oval nucleus of the bed nucleus of the stria terminalis (BSTov) have been linked closely anatomically and functionally. To determine whether these regions may be subdivided further on a neurochemical basis, dual in situ hybridization was used to determine the colocalization of corticotropin-releasing hormone (CRH), enkephalin (ENK), or neurotensin (NT) with glutamic acid decarboxylase isoforms 65 and 67 [used concurrently as a marker for gamma-aminobutyric acid GABA] in these nuclei. It was found that, for both regions, each peptide invariably was localized in a GABAergic cell. Although there was a similar overlap in the distribution of NT with ENK in the BSTov and CEAl, it was observed that CRH and ENK rarely were colocalized in either nucleus. To determine whether these distinct neuronal populations could be activated differentially, male rats were given a systemic injection of interleukin-1beta (IL-1beta; 5 microg/kg, i.p.), a stimulus that results in a robust increase in c-fos mRNA expression in the BSTov and CEAl. The neurochemical identity of these activated neurons showed striking similarities between the BSTov and the CEAl; All IL-1beta-responsive cells were GABAergic, the majority of c-fos- positive cells expressed ENK mRNA (BSTov, 81%; CEAl, 94%), and some expressed NT mRNA (BSTov, 23%; CEAl, 22%), whereas very few expressed CRH mRNA (BSTov, 4%; CEAl, 1%). These data provide evidence for the existence of discrete neural circuits within the BSTov and CEAl, and the similarities in the patterns of neurochemical colocalization in these nuclei are consistent with the concept of an extended amygdala. Furthermore, these data indicate that intraperitoneal IL-1beta recruits neurochemically distinct pathways within the BSTov and CEAl, and it is suggested that this differential activation may mediate specific aspects of immune, limbic, and/or autonomic processes. PMID- 10464375 TI - Central afferent pathways conveying nociceptive input to the hypothalamic paraventricular nucleus as revealed by a combination of retrograde labeling and c fos activation. AB - Previous data have shown that noxious thermal stimulation of the hind leg in the anesthetized rat causes c-fos activation in the paraventricular nucleus of the hypothalamus (PVN); in other brain nuclei, including the cathecholaminergic cell groups of the caudal medulla; and in the adenohypophysis. Stimulation was followed by adrenocorticotropic hormone plasma release but did not provoke cardiovascular changes. In the current study, the afferent central pathways conveying the nociceptive input to the PVN were studied throughout the brain by using double labeling for the Fos-protein and the retrograde tracer cholera toxin subunit B (CTb) injected into the PVN. Although double labeling occurred in several hypothalamic nuclei, the periaqueductal gray, the lateral parabrachial area, and the catecholaminergic medullary groups, high rates of double labeling occurred only in the cells of the A1 region of the ventrolateral medulla ( approximately 83% of CTb-labeled cells expressing c-fos). Further triple labeling with tyrosine hydroxylase (TH) revealed that > 80% of the double-labeled cells were TH-immunoreactive. The spinal cord had the usual strong c-fos expression but showed no retrograde labeling from the PVN. Noxious stimulation caused corticosterone plasma release. To ascertain a possible link of spinofugal neurons with the A1 cells, biotinylated dextran amine was injected into the spinal dorsal horn. Numerous anterogradely labeled fibers with bouton-like structures were observed, with the latter apposed to double- and triple-labeled cells in the A1 region. It is suggested that a dysynaptic route relayed in the A1 region conveys the nociceptive somatic input from the spinal cord to the PVN. Noxious stimulation may act as a systemic stressor, activating the hypothalamic-pituitary adrenal axis. PMID- 10464376 TI - Rapid extension of axons into the CA3 region by adult-generated granule cells. AB - The dentate gyrus continues to produce granule neurons throughout adulthood. The present study examined the extension of axons by adult-generated granule neurons into hippocampal area CA3. We injected the fluorescent retrograde tracers Fast blue (FB) and FluoroRuby (FR) into area CA3 of adult male rats at various times after the administration of 5'-bromo-2'-deoxyuridine (BrdU), a marker of proliferating cells and their progeny. We report that immature granule cells extend axons into CA3 as rapidly as 4-10 days after mitosis. A significant increase in the percentage of BrdU-labeled cells that were labeled with FB or FR was observed by 2 weeks after BrdU administration. This proportion remained roughly constant up to 3 weeks after BrdU-labeling, a time at which markers of a mature neuronal phenotype are expressed. BrdU-labeled cells that contained either FB or FR often were located far from the tracer injection site, indicating that these cells had extended relatively long axons. Collectively these results suggest that adult-generated granule neurons may influence normal hippocampal function, even at a very early stage after their production. PMID- 10464377 TI - Indoleamine-accumulating amacrine cells are presynaptic to rod bipolar cells through GABA(C) receptors. AB - gamma-Aminobutyric acid (GABA), is a main source of inhibitory modulation of the rod pathway in the mammalian retina. The authors previously showed that rod bipolar cells express at least three types of ionotropic GABA receptors. Here, the authors sought to determine which neurons are the presynaptic partners at these synapses in the rabbit retina. Indoleamine-accumulating amacrine cells (IACs) were immunolabeled with an antiserum against serotonin (5HT) in vertical sections and wholemounts of rabbit retinae that had been preloaded with 5HT. The tissue was double labeled for the rho subunits of the GABA(C) receptor or the alpha3 subunit of the GABA(A) receptor. Punctate immunofluorescence was observed for both receptor subunits and was found to coincide with the dendrites and varicosities of IACs. The localization of rho subunits was examined at the ultrastructural level by using postembedding techniques on slam-frozen, cryosubstituted tissue. Double labeling at the electron microscopic level revealed that 5HT-immunoreactive processes were presynaptic to rod bipolar cells through GABA(C) receptors. Intracellular injection of the two morphologic subclasses of IAC amacrine cells, S1 and S2, with Lucifer yellow followed by immunolabeling for the alpha3 or rho subunits revealed that varicosities on the dendrites of both cell types were in register with alpha3- and rho-immunoreactive puncta. Taken together, these results suggest that IACs are presynaptic to rod bipolar cells through GABA(C) receptors and possibly through GABA(A) receptors. PMID- 10464378 TI - p53 and Bax: putative death factors in taste cell turnover. AB - The turnover of cells in renewing epithelia presents an opportunity to examine cell death pathways in adult vertebrates. In mouse lingual epithelium a typical taste receptor cell survives for 9 days, until it is killed by an unknown cascade of death factors. Apoptosis was implicated by the presence of fragmented DNA in about 8% of taste receptor cells in the vallate papilla. In using immunocytochemistry to seek putative death factors, we observed that squamous epithelial cells of the tongue were negative for Bax, a death factor in the Bcl-2 family of survival/death factors, and were also negative for p53, a tumor suppressor protein linked to apoptosis and Bax transcription. In contrast, 8-10% of the taste receptor cells were Bax-positive, and 9-11% were p53 positive. These immunopositive taste receptor cells were more likely to display death-related morphologic defects than other receptor cells, and they frequently coexpressed p53 and Bax. In both neonatal and adult mice, the labeling of dividing cells with 5-bromo-2'-deoxyuridine indicated that all Bax-positive taste cells were at least 5 days old. On postnatal day 7, when few taste cells were old, no more than 1% of taste cells were immunopositive for either p53 or Bax. We inferred that old taste receptor cells employ p53 and Bax as part of their apoptotic death pathway. The routine expression of p53 by postmitotic, aged taste cells broadens the conventional view that p53 is restricted to mitotic cells that have stress damaged DNA. Furthermore, the scattered distribution of aged receptor cells within the taste bud excludes some explanations for stable taste signals during receptor cell turnover. PMID- 10464379 TI - The regulation of apoptotic cell death. AB - Apoptosis is a fundamental biological phenomenon in which the death of a cell is genetically and biochemically regulated. Different molecules are involved in the regulation of the apoptotic process. Death receptors, coupled to distinct members of the caspases as well as other adapter molecules, are involved in the initiation of the stress signals (The Indictment). Members of the Bcl-2 family control at the mitochondrial level the decision between life and death (The Judgement). The effector caspases are responsible for all morphological and biochemical changes related to apoptosis including the "eat-me" signals perceived by phagocytes and neighboring cells (The Execution). Finally, apoptosis would have little biological significance without the recognition and removal of the dying cells (The Burial). PMID- 10464380 TI - Photodynamic DNA damage induced by phycocyanin and its repair in Saccharomyces cerevisiae. AB - In the present study, we analyzed DNA damage induced by phycocyanin (PHY) in the presence of visible light (VL) using a set of repair endonucleases purified from Escherichia coli. We demonstrated that the profile of DNA damage induced by PHY is clearly different from that induced by molecules that exert deleterious effects on DNA involving solely singlet oxygen as reactive species. Most of PHY induced lesions are single strand breaks and, to a lesser extent, base oxidized sites, which are recognized by Nth, Nfo and Fpg enzymes. High pressure liquid chromatography coupled to electrochemical detection revealed that PHY photosensitization did not induce 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) at detectable levels. DNA repair after PHY photosensitization was also investigated. Plasmid DNA damaged by PHY photosensitization was used to transform a series of Saccharomyces cerevisiae DNA repair mutants. The results revealed that plasmid survival was greatly reduced in rad14 mutants, while the ogg1 mutation did not modify the plasmid survival when compared to that in the wild type. Furthermore, plasmid survival in the ogg1 rad14 double mutant was not different from that in the rad14 single mutant. The results reported here indicate that lethal lesions induced by PHY plus VL are repaired differently by prokaryotic and eukaryotic cells. Moreover, nucleotide excision repair seems to play a major role in the recognition and repair of these lesions in Saccharomyces cerevisiae. PMID- 10464381 TI - McFarland nephelometer as a simple method to estimate the sensitivity of the polymerase chain reaction using Mycobacterium tuberculosis as a research tool. AB - Polymerase chain reaction (PCR) has been widely investigated for the diagnosis of tuberculosis. However, before this technique is applied on clinical samples, it needs to be well standardized. We describe the use of McFarland nephelometer, a very simple approach to determine microorganism concentration in solution, for PCR standardization and DNA quantitation, using Mycobacterium tuberculosis as a model. Tuberculosis is an extremely important disease for the public health system in developing countries and, with the advent of AIDS, it has also become an important public health problem in developed countries. Using Mycobacterium tuberculosis as a research model, we were able to detect 3 M. tuberculosis genomes using the McFarland nephelometer to assess mycobacterial concentration. We have shown here that McFarland nephelometer is an easy and reliable procedure to determine PCR sensitivity at lower costs. PMID- 10464382 TI - Phenotyping and genotyping methods applied to investigate the relatedness of Brazilian isolates of Enterobacter cloacae. AB - In order to evaluate the resolving power of several typing methods to identify relatedness among Brazilian strains of Enterobacter cloacae, we selected twenty isolates from different patients on three wards of a University Hospital (Orthopedics, Nephrology, and Hematology). Traditional phenotyping methods applied to isolates included biotyping, antibiotic sensitivity, phage-typing, and O-serotyping. Plasmid profile analysis, ribotyping, and macrorestriction analysis by pulsed-field gel electrophoresis (PFGE) were used as genotyping methods. Sero- and phage-typing were not useful since the majority of isolates could not be subtyped by these methods. Biotyping, antibiogram and plasmid profile permitted us to classify the samples into different groups depending on the method used, and consequently were not reliable. Ribotyping and PFGE were significantly correlated with the clinical epidemiological analysis. PFGE did not type strains containing nonspecific DNase. Ribotyping was the most discriminative method for typing Brazilian isolates of E. cloacae. PMID- 10464383 TI - Infrequent p53 gene alterations in ulcerative colitis. AB - The purpose of this study was to determine whether point mutations and loss of the p53 gene take place in ulcerative colitis which is histologically negative for dysplasia. DNA was extracted from 13 frozen rectal or colon biopsies and blood samples. Ulcerative colitis was classified histologically as active (10 cases) and inactive (3 cases). Exons 5-8 were amplified by PCR, treated with exonuclease and shrimp alkaline phosphatase and sequenced by the dideoxy chain termination method with the Sequenase Version 2.0 DNA sequencing kit. PCR products of intron 6 and exon 4 were digested with MspI and AccII, respectively, for RFLP analysis. No p53 gene mutation was detected in these cases. The number of informative patients for loss of heterozygosity (LOH) at the p53 intron 6 was high, 11 out of 12 (92%), whereas no LOH was observed. LOH affecting p53 exon 4 was not detected in lesions from 5 of 12 patients (42%). In ulcerative colitis, tumor progression is similar to that in sporadic colon cancer, and other oncogenes and tumor suppressor genes are likely to be mutated before the p53 gene. PMID- 10464384 TI - A gasometric method to determine erythrocyte catalase activity. AB - We describe a new gasometric method to determine erythrocyte catalase activity by the measurement of the volume of oxygen produced as a result of hydrogen peroxide decomposition in a system where enzyme and substrate are separated in a special reaction test tube connected to a manometer and the reagents are mixed with a motor-driven stirrer. The position of the reagents in the test tube permits the continuous measurement of oxygen evolution from the time of mixing, without the need to stop the reaction by the addition of acid after each incubation time. The enzyme activity is reported as K Hb, i.e., mg hydrogen peroxide decomposed per second per gram of hemoglobin (s-1 g Hb-1). The value obtained for catalase activity in 28 samples of hemolyzed human blood was 94.4 +/- 6.17 mg H2O2 s-1 g Hb-1. The results obtained were precise and consistent, indicating that this rapid, simple and inexpensive method could be useful for research and routine work. PMID- 10464385 TI - Disseminated Langerhans' cell histiocytosis and massive protein-losing enteropathy. AB - Symptomatic involvement of the gastrointestinal (GI) tract as a prominent symptom in Langerhans' cell histiocytosis (LCH) is uncommon, occurring in less than 1 to 5% of all cases, even when the disease is in its disseminated form. Up to now, there have been reports of 18 cases of LCH with GI manifestations, including our 2 cases, with diarrhea (77.7%), protein-losing enteropathy (33.3%) and bloody stool being the most frequent findings. The authors present two patients with severe diarrhea and refractory hypoalbuminemia, and with the protein-losing enteropathy documented by Cr51-labeled albumin studies. A review of the literature indicated that the presence of GI symptoms is often associated with systemic disease as well as with poor prognosis, mainly under 2 years of age. Radioisotopes are useful for documenting protein loss in several diseases with high specificity and sensitivity, and their utilization in the cases reviewed here permitted diagnoses in 6 children, as well as improved therapeutic management. PMID- 10464386 TI - Detection of specific IgE antibodies in parasite diseases. AB - Activation of Th1 or Th2 cells is associated with production of specific immunoglobulin isotypes, offering the opportunity to use antibody measurement for evaluation of T cell function. Schistosomiasis and visceral leishmaniasis are diseases associated with Th2 activation. However, an IgE response is not always detected in these patients. In the present study we evaluated specific IgE antibodies to S. mansoni and L. chagasi antigens by ELISA after depletion of serum IgG with protein G immobilized on Sepharose beads or RF-absorbent (purified sheep IgG antibodies anti-human IgG). In schistosomiasis patients, specific IgE to SWAP antigen was demonstrable in only 10 of 21 patients (48%) (mean absorbance +/- SD = 0.102 +/- 0.195) when unabsorbed serum was used. Depletion of IgG with protein G increased the number of specific IgE-positive tests to 13 (62%) and the use of RF-absorbent increased the number of positive results to 20 (95%) (mean absorbances +/- SD = 0.303 +/- 0. 455 and 0.374 +/- 0.477, respectively). Specific IgE anti-L. chagasi antibodies were not detected in unabsorbed serum from visceral leishmaniasis patients. When IgG was depleted with protein G, IgE antibodies were detected in only 3 (11%) of 27 patients, and the use of RF absorbent permitted the detection of this isotype in all 27 visceral leishmaniasis sera tested (mean absorbance +/- SD = 0.104 +/- 0.03). These data show that the presence of IgG antibodies may prevent the detection of a specific IgE response in these parasite diseases. RF-absorbent, a reagent that blocks IgG binding sites and also removes rheumatoid factor, was more efficient than protein G for the demonstration of specific IgE antibodies. PMID- 10464387 TI - Prevalence and risk factors for HBV, HCV and HDV infections among injecting drug users from Rio de Janeiro, Brazil. AB - Viral hepatitis constitutes a major health issue, with high prevalence among injecting drug users (IDUs). The present study assessed the prevalence and risk determinants for hepatitis B, C and D viruses (HBV, HCV and HDV) infections among 102 IDUs from Rio de Janeiro, Brazil. Serological markers and HCV-RNA were detected by enzyme immunoassay and nested PCR, respectively. HCV genotyping was determined by restriction fragment length polymorphism analysis (RFLP). HBsAg, anti-HBc and anti-HBs were found in 7.8, 55.8 and 24. 7% of IDUs, respectively. In the final logistic regression, HBV infection was independently associated with male homosexual intercourse within the last 5 years (odds ratio (OR) 3.1; 95% confidence interval (CI) 1.1-8.8). No subject presented anti-delta (anti-HD). Anti-HCV was detected in 69.6% of subjects, and was found to be independently associated with needle sharing in the last 6 months (OR 3.4; 95% CI 1.3-9.2) and with longer duration of iv drug use (OR 3.1; 95% CI 1.1-8.7). These data demonstrate that this population is at high risk for both HBV and HCV infection. Among IDUs from Rio de Janeiro, unprotected sexual intercourse seems to be more closely associated with HBV infection, whereas HCV is positively correlated with high risk injecting behavior. Comprehensive public health interventions targeting this population and their sexual partners must be encouraged. PMID- 10464388 TI - Probable effect of photoperiod on seasonal variation in the nuclear volume of the adrenal cortex of viscacha (Lagostomus maximus maximus). AB - The neuroendocrine system regulates several organic functions such as reproduction, metabolism and adaptation to the environment. This system shows seasonal changes linked to the environment. The experimental model used in the present study was Lagostomus maximus maximus (viscacha). The reproduction of males of this species is photoperiod dependent. Twenty-four adult male viscachas were captured in their habitat at different times during one year. The adrenal glands were processed for light microscopy. Serial cuts were stained with hematoxylin-eosin for the morphometric study, and 100 nuclei of each zone of the adrenal cortex were counted per animal. Data were analyzed statistically by ANOVA and the Tukey test. The cells of the glomerulosa zone are arranged in a tube shaped structure. The fasciculata zone has large cells with central nuclei and clearly visible nucleoli and with a vacuolar cytoplasm. In the reticularis zone there are two of types of cells, one with a nucleus of fine chromatin and a clearly visible nucleolus and the other with nuclear pycnosis. Morphometric analysis showed maximum nuclear volumes during the February-March period with values of 133 +/- 7.3 microm3 for the glomerulosa, 286.4 +/- 14.72 microm3 for the fasciculata, and 126.3 +/- 9.49 microm3 for the reticularis. Minimum nuclear volumes were observed in August with values of 88.24 +/- 9.9 microm3 for the glomerulosa, 163.7 +/- 7.78 microm3 for the fasciculata and 64.58 +/- 4.53 microm3 for the reticularis. The short winter photoperiod to which viscacha is subjected could inhibit the adrenal cortex through a melatonin increase which reduces the nuclear volume as well as the cellular activity. PMID- 10464389 TI - The relationship between anxiety and depression in animal models: a study using the forced swimming test and elevated plus-maze. AB - The present study evaluated the correlation between the behavior of mice in the forced swimming test (FST) and in the elevated plus-maze (PM). The effect of the order of the experiments, i.e., the influence of the first test (FST or PM) on mouse behavior in the second test (PM or FST, respectively) was compared to handled animals (HAND). The execution of FST one week before the plus-maze (FST PM, N = 10), in comparison to mice that were only handled (HAND-PM, N = 10) in week 1, decreased % open entries (HAND-PM: 33.6 +/- 2.9; FST-PM: 20.0 +/- 3.9; mean +/- SEM; P<0.02) and % open time (HAND-PM: 18.9 +/- 3.3; FST-PM: 9.0 +/- 1.9; P<0.03), suggesting an anxiogenic effect. No significant effect was seen in the number of closed arm entries (FST-PM: 9.5 (7.0-11.0); HAND-PM: 10.0 (4.0 14.5), median (interquartile range); U = 46.5; P>0.10). A prior test in the plus maze (PM-FST) did not change % immobility time in the FST when compared to the HAND-FST group (HAND-FST: 57.7 +/- 3.9; PM-FST: 65.7 +/- 3.2; mean +/- SEM; P>0.10). Since these data suggest that there is an order effect, the correlation was evaluated separately with each test sequence: FST-PM (N = 20) and PM-FST (N = 18). There was no significant correlation between % immobility time in the FST and plus-maze indexes (% time and entries in open arms) in any test sequence (r: 0.07 to 0.18). These data suggest that mouse behavior in the elevated plus-maze is not related to behavior in the forced swimming test and that a forced swimming test before the plus-maze has an anxiogenic effect even after a one-week interval. PMID- 10464390 TI - Role of the hippocampus in contextual memory after classical aversive conditioning in pigeons (C. livia). AB - We investigated the effects of hippocampal lesions with ibotenic acid (IBO) on the memory of the sound-context-shock association during reexposure to the conditioning context. Twenty-nine adult pigeons were assigned to a non-lesioned control group (CG, N = 7), a sham-lesioned group (SG, N = 7), a hippocampus lesioned experimental group (EG, N = 7), and to an unpaired nonlesioned group (tone-alone exposure) (NG, N = 8). All pigeons were submitted to a 20-min session in the conditioning chamber with three associations of sound (1000 Hz, 85 dB, 1 s) and shock (10 mA, 1 s). Experimental and sham lesions were performed 24 h later (EG and SG) when EG birds received three bilateral injections (anteroposterior (A), 4.5, 5.25 and 7.0) of IBO (1 microl and 1 microg/microl) and SG received one bilateral injection (A, 5.25) of PBS. The animals were reexposed to the training context 5 days after the lesion. Behavior was videotaped for 20 min and analyzed at 30-s intervals. A significantly higher percent rating of immobility was observed for CG (median, 95.1; range, 79.2 to 100.0) and SG (median, 90.0; range, 69.6 to 95.0) compared to EG (median, 11.62; range, 3.83 to 50.1) and NG (median, 7.33; range, 6.2 to 28.1) (P<0.001) in the training context. These results suggest impairment of contextual fear in birds who received lesions one day after conditioning and a role for the hippocampus in the modulation of emotional aversive memories in pigeons. PMID- 10464391 TI - Diagnosing dementia and normal aging: clinical relevance of brain ratios and cognitive performance in a Brazilian sample. AB - The main objective of the present study was to evaluate the diagnostic value (clinical application) of brain measures and cognitive function. Alzheimer and multi-infarct patients (N = 30) and normal subjects over the age of 50 (N = 40) were submitted to a medical, neurological and cognitive investigation. The cognitive tests applied were Mini-Mental, word span, digit span, logical memory, spatial recognition span, Boston naming test, praxis, and calculation tests. The brain ratios calculated were the ventricle-brain, bifrontal, bicaudate, third ventricle, and suprasellar cistern measures. These data were obtained from a brain computer tomography scan, and the cutoff values from receiver operating characteristic curves. We analyzed the diagnostic parameters provided by these ratios and compared them to those obtained by cognitive evaluation. The sensitivity and specificity of cognitive tests were higher than brain measures, although dementia patients presented higher ratios, showing poorer cognitive performances than normal individuals. Normal controls over the age of 70 presented higher measures than younger groups, but similar cognitive performance. We found diffuse losses of tissue from the central nervous system related to distribution of cerebrospinal fluid in dementia patients. The likelihood of case identification by functional impairment was higher than when changes of the structure of the central nervous system were used. Cognitive evaluation still seems to be the best method to screen individuals from the community, especially for developing countries, where the cost of brain imaging precludes its use for screening and initial assessment of dementia. PMID- 10464392 TI - Effects of diazepam on Mycobacterium bovis-induced infection in hamsters. AB - The in utero exposure of hamsters to low doses of diazepam results in impaired host defense against Mycobacterium bovis during adulthood. Delayed developmental immunotoxicity, however, represents a specific situation that might not be general. The present experiment was undertaken to investigate the effects of diazepam on hamster resistance to M. bovis using adult animals. The effects of diazepam treatment on serum cortisol levels were also studied. Adult hamsters (N = 10 for each group) were treated with diazepam (E1 = 1. 0, E2 = 2.0 or E3 = 3.0 mg kg-1 day-1 subcutaneously) or with control solution (C) for 30 days. Seven days after the beginning of the treatment, the animals received identical inoculum concentrations of M. bovis. Hamsters treated with the higher (2.0 and 3.0 mg kg-1 day-1) doses of diazepam exhibited: 1) increased granuloma areas in the liver (C = 1.81 +/- 1.39, E2 = 10.29 +/- 4.64 and E3 = 15.80 +/- 4.82) and lung (C = 0.54 +/- 0.55, E2 = 6.28 +/- 3.85 and E3 = 6.31 +/- 3.56) and 2) increased scores of M. bovis colony-forming units isolated from liver (C = 2.0, E2 = 3.0 and E3 = 3.5), lung (C = 1.0, E2 = 3.0 and E3 = 3.5) and spleen (C = 1.0, E2 = 2.5 and E3 = 4.0). These effects were dose dependent, and were not detected or were less severe in animals treated with the lowest (1.0 mg/kg) dose of diazepam as well as in those of the control group. Furthermore, diazepam treatment (3.0 mg kg-1 day-1 for 30 days) increased (E3 = 71.32 +/- 2.99; N = 10) the serum levels of cortisol compared to control hamsters (C = 22.61 +/- 2.75; N = 10). The present data, that demonstrate an impaired defense against M. bovis in adult hamsters treated with diazepam, were tentatively explained on the basis of a direct and/or indirect action of diazepam on the cytokine network. The effects may be related to stimulation of peripheral benzodiazepine receptor binding sites (PBR) by macrophages and/or lymphocytes, or they may be mediated by PBR stimulation of the adrenals. PMID- 10464393 TI - Interaction between substance P and gastrin-releasing peptide on thyrotropin secretion by rat pituitary in vitro. AB - The effect of substance P (SP) on thyrotropin (TSH) secretion is controversial. In this study we evaluated the effect of SP on TSH secretion by hemipituitaries of 3-month-old Wistar rats in vitro and its interaction with gastrin-releasing peptide (GRP) at equimolar concentrations (1 microM and 10 microM). TSH release was measured under basal conditions and 30 min after incubation in the absence or presence of SP, GRP or both peptides. Pituitary TSH content was also measured in the pituitary homogenate after incubation. SP at both concentrations caused a significant (P<0.05) increase in TSH secretion compared with all other groups, which was approximately 60% (1 microM) and 85% (10 microM) higher than that of the control group (23.3 +/- 3.0 ng/ml). GRP at the lower concentration did not produce a statistically significant change in TSH secretion, whereas at the concentration of 10 microM it produced a 50% reduction in TSH. GRP co-incubated with substance P completely blocked the stimulatory effect of SP at both concentrations. Pituitary TSH content decreased in the SP-treated group compared to controls (0.75 +/- 0.03 microg/hemipituitary) at the same proportion as the increase in TSH secretion, and this effect was also blocked when GRP and SP were co-incubated. In conclusion, in an in vitro system, SP increased TSH secretion acting directly at the pituitary level and this effect was blocked by GRP, suggesting that GRP is more potent than SP on TSH secretion, and that this inhibitory effect could be the predominant effect in vivo. PMID- 10464394 TI - Antisense mRNA for NPY-Y1 receptor in the medial preoptic area increases prolactin secretion. AB - We investigated the participation of neuropeptide Y-Y1 receptors within the medial preoptic area in luteinizing hormone, follicle-stimulating hormone and prolactin release. Four bilateral microinjections of sense (control) or antisense 18-base oligonucleotides of messenger ribonucleic acid (mRNA) (250 ng) corresponding to the NH2-terminus of the neuropeptide Y1 receptor were performed at 12-h intervals for two days into the medial preoptic area of ovariectomized Wistar rats (N = 16), weighing 180 to 200 g, treated with estrogen (50 microg) and progesterone (25 mg) two days before the experiments between 8.00 and 10:00 a.m. Blockade of Y1 receptor synthesis in the medial preoptic area by the antisense mRNA did not change plasma luteinizing hormone or follicle-stimulating hormone but did increase prolactin from 19.6 +/- 5.9 ng/ml in the sense group to 52.9 +/- 9.6 ng/ml in the antisense group. The plasma hormones were measured by radioimmunoassay and the values are reported as mean +/- SEM. These data suggest that endogenous neuropeptide Y in the medial preoptic area has an inhibitory action on prolactin secretion through Y1 receptors. PMID- 10464395 TI - Possible involvement of A1 receptors in the inhibition of gonadotropin secretion induced by adenosine in rat hemipituitaries in vitro. AB - We investigated the participation of A1 or A2 receptors in the gonadotrope and their role in the regulation of LH and FSH secretion in adult rat hemipituitary preparations, using adenosine analogues. A dose-dependent inhibition of LH and FSH secretion was observed after the administration of graded doses of the R isomer of phenylisopropyladenosine (R-PIA; 1 nM, 10 nM, 100 nM, 1 microM and 10 microM). The effect of R-PIA (10 nM) was blocked by the addition of 8 cyclopentyltheophylline (CPT), a selective A1 adenosine receptor antagonist, at the dose of 1 microM. The addition of an A2 receptor-specific agonist, 5-N methylcarboxamidoadenosine (MECA), at the doses of 1 nM to 1 microM had no significant effect on LH or FSH secretion, suggesting the absence of this receptor subtype in the gonadotrope. However, a sharp inhibition of the basal secretion of these gonadotropins was observed after the administration of 10 microM MECA. This effect mimicked the inhibition induced by R-PIA, supporting the hypothesis of the presence of A1 receptors in the gonadotrope. R-PIA (1 nM to 1 microM) also inhibited the secretion of LH and FSH induced by phospholipase C (0.5 IU/ml) in a dose-dependent manner. These results suggest the presence of A1 receptors and the absence of A2 receptors in the gonadotrope. It is possible that the inhibition of LH and FSH secretion resulting from the activation of A1 receptors may have occurred independently of the increase in membrane phosphoinositide synthesis. PMID- 10464396 TI - Contribution to the sand fly fauna (Diptera: Phlebotominae) of Rio Grande do Sul, Brazil and Leishmania (Viannia) infections. AB - American cutaneous leishmaniasis (ACL) is endemic in the State of Rio Grande do Sul, Brazil. As reports of nearby hospitals suggest, the Parque Estadual do Turvo seems to serve as a source for the disease. During three months from November 1996 to February 1997 we collected, in this park, 2,228 sandflies (10 Lutzomyia species and 2 6species). We applied the polymerase chain reaction to 920 females which belong to the following species: Lutzomyia migonei, Lu. pessoai, Lu. fischeri, Lu. misionensis, Lu. lanei, Lu. neivai, Lu. shannoni, and Lu. monticola, in an attempt to verify natural infection by Leishmania (Viannia), the causative agent of ACL. Le. (Viannia) infections were demonstrated by DNA amplification from two Lu. pessoai and one Lu. misionensis female. Lu. pessoai have been found with leptomonas in the gut believed to be Le. (V.) braziliensis in other endemic areas of northeastern and southeastern Brazil. However, Lu. misionensis has never been found carrying a natural infection of Le. (Viannia). PMID- 10464397 TI - Epidemiology of hookworm infection in Itagua, Paraguay: a cross sectional study. AB - A cross-sectional study in Itagua, Paraguay tested 192 people for the presence, intensity and species of hookworm infection. Fifty-nine percent of these individuals were found to be infected. Intensity of infection was determined on 92% of infected individuals by quantitative egg counts. The high intensity hookworm infections, which cause the greatest morbidity, were clustered between the ages of five and 14 years. No differences were seen between genders. The species of hookworm was determined for parasites reared from 72% of infected individuals. Both Necator americanus and Ancylostoma duodenale were identified, although the former species predominated. We conclude that hookworm infection continues to be a public health problem in Paraguay, particularly among children and adolescents who suffer from high intensity infections. A. duodenale continues to persist in the Western Hemisphere and has not been completely displaced by N. americanus. PMID- 10464398 TI - Survey of canine heartworm in the city of Recife, Pernambuco, Brazil. AB - Six hundred and eleven random-source dogs (338 male, 273 female) one year of age or older, from six sections of the city of Recife, Pernambuco, were examined antemortem for circulating microfilariae Dirofilaria immitis and Dipetalonema reconditum adult heartworm (D. immitis) antigen, and examined postmortem for adult heartworms. The prevalence of heartworm infection was 2.3% (14/611), as determined by necropsy for adult worms, and 1% (6/611) had circulating microfilariae of D. immitis; thus, 57.1% of the heartworm-infected dogs had occult infections. The results of serological testing indicated that 1.3% (8/611) of the dogs were positive for adult heartworm antigen. A total of 42 (6.9%) of the dogs had microfilariae of D. reconditum; 40 of these had only D. reconditum and two additional dogs had microfilariae of both species, D. immitis and D. reconditum. PMID- 10464399 TI - Bloodmeal microfilariae density and the uptake and establishment of Wuchereria bancrofti infections in Culex quinquefasciatus and Aedes aegypti. AB - The relationship between ingestion of microfilariae (mf), production of infective larvae (L3) and mf density in human blood has been suggested as an important determinant in the transmission dynamics of lymphatic filariasis. Here we assess the role of these factors in determining the competence of a natural vector Culex quinquefasciatus and a non vector Aedes aegypti to transmit Wuchereria bancrofti. Mosquitoes were infected via a membrane feeding procedure. Both mosquito species ingested more than the expected number of microfilariae (concentrating factor was 1.28 and 1.81 for Cx. quinquefasciatus and Ae. aegypti, respectively) but Cx. quinquefasciatus ingested around twice as many mf as Ae. aegypti because its larger blood meal size. Ae. aegypti showed a faster mf migration capacity compared to Cx. quinquefasciatus but did not allow parasite maturation under our experimental conditions. Similar proportions of melanized parasites were observed in Ae. aegypti (2. 4%) and Cx. quinquefasciatus (2.1%). However, no relationship between rate of infection and melanization was observed. We conclude that in these conditions physiological factors governing parasite development in the thorax may be more important in limiting vectorial competence than the density of mf ingested. PMID- 10464400 TI - Parasitism of Ixodes (Multidentatus) auritulus Neumann (Acari: Ixodidae) on birds from the city of Curitiba, State of Parana, Southern Brazil. AB - The tick-bird relationship of 56 specimens of birds (Passeriformes and Columbiformes) collected in the city of Curitiba, State of Parana, between 1990 and 1995, among which 102 specimens of Ixodes (Multidentatus) auritulus were found and analyzed. New host records were also produced including the first report of I. auritulus on a Columbiformes bird in Brazil. PMID- 10464401 TI - Infections by helminth parasites in "puyenes", Galaxias maculatus (Galaxiidae, Salmoniformes), from Southern Argentina with special reference to Tylodelphys barilochensis (Digenea, Platyhelminthes). AB - The occurrence of Tylodelphys barilochensis, Acanthostomoides apophalliformis, Contracaecum sp. and Camallanus corderoi infecting Galaxias maculatus ("puyenes") was quantified for the first time in Lake Nahuel Huapi, southern Argentina. T. barilochensis was recorded in this lake for the first time. The role of G. maculatus population in transmission of parasites to the salmonids is more important for Contracaecum sp. (prevalence 14-34%) and A. apophalliformis (prevalence 30-54%) than for C. corderoi (prevalence 6-8%). The absence of Diphyllobothrium spp. in samples shows that the G. maculatus population does not play any role in the life cycles of these important zoonotic parasites. The sex of the host had no effect on T. barilochensis abundance. Statistical differences in T. barilochensis abundance between "puyenes" of the same size class between sampling stations and positive correlation between prevalence of infected snails and T. barilochensis abundance in fish suggest that different stocks have been sampled. Factors influencing T. barilochensis abundance are discussed. PMID- 10464402 TI - Intestinal parasites of some diarrhoeic HIV-seropositive individuals in North Brazil, with particular reference to Isospora belli Wenyon, 1923 and Dientamoeba fragilis Jepps & Dobell, 1918. PMID- 10464403 TI - Redescription of Rhamnocercus stichospinus Seamster and Monaco, 1956 (Monogenea: Diplectanidae), parasitic on Menticirrhus americanus (Osteichthyes: Sciaenidae) from the coastal zone of the State of Rio de Janeiro, Brazil. AB - Rhamnocercus stichospinus Seamster and Monaco, 1956 (Diplectanidae) parasitic on the sciaenid fish Menticirrhus americanus from the coastal zone of the State of Rio de Janeiro, is redescribed and recorded for the first time in the South American Atlantic Ocean. The generic diagnosis of Rhamnocercus is emended to accommodate the presence of confluent intestinal ceca in R. stichospinus. PMID- 10464404 TI - Description of male, larva and pupa of Stibasoma theotaenia (Wiedemann) (Diptera Tabanidae). AB - Unknown male, larva and pupa of Stibasoma theotaenia from northern Argentina are described and illustrated. Larvae were collected from terrestrial Bromeliaceae of Aechnea sp. and maintained in the laboratory through development. PMID- 10464405 TI - The type specimens of sucking lice (Anoplura) deposited in the entomological collection of Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brazil. AB - This study presents a list of 34 Anoplura type specimens deposited in the Werneck Collection of Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brazil. It includes 18 holotypes, 16 allotypes, 88 paratypes and 10 neotypes, distributed among the genera: Enderleinellus, Fahrenholzia, Haematopinus, Hoplopleura, Linognathus, Microthoracius, Pecaroecus, Polyplax and Pterophthirus. The types are related according to their respective data and literature. PMID- 10464406 TI - Lutzomyia derelicta (Diptera: Psychodidae) a singular new phlebotomine sand fly from an Inselberg in Northeastern Amazonia. AB - Lutzomyia derelicta n. sp. is described from specimens collected in an isolated xeric habitat in the rainforest in the north of the State of Para, Brazil. The new species lacks the posterior bulge in the dorsal wall of the cibarium characteristic of the New World genus Lutzomyia, and the armature of the male genitalia is of the pattern found elsewhere only in the Old World species of Sergentomyia. L. derelicta is phenetically intermediate between the known species of Lutzomyia and Sergentomyia, and cannot readily be placed in any existing subgenus or species group of either genus. PMID- 10464407 TI - Studies on Procamallanus (Spirocamallanus) Pereirai annereaux, 1946 (Nematoda: Camallanidae), with new host records and new morphological data on the larval stages. AB - Larval stages and adults of Procamallanus (Spirocamallanus) pereirai Annereaux, 1946 are described from naturally infected Paralonchurus brasiliensis (Steindachner) (Sciaenidae) from the coast of the State of Rio de Janeiro, Brazil. The translucent first-stage larvae have a denticulate process at the anterior end, no buccal capsule or esophagus undifferentiated into anterior muscular and posterior glandular parts and an elongate tail; third-stage larvae have a tail with three terminal projections, a buccal capsule divided into an anterior portion with 12-20 ridges running to the left and a posterior smooth portion, and an esophagus with muscular and glandular regions. Fourth-stage larvae exhibit a buccal capsule lacking a distinct basal ring with ridges running to the right and a tail with two terminal processes, as in adults. New host records are reported and their role in its life-cycle are discussed. PMID- 10464408 TI - Leishmania panamensis: a 44bp deletion in gp63 gene is found in cDNA and genomic libraries. PMID- 10464409 TI - Experimental infection of canine peritoneal macrophages with visceral and dermotropic Leishmania strains. AB - A study was carried out using macrophages cultured from the peritoneal exudate of dogs infected in vitro with three species of Leishmania: L. (L.) chagasi, L. (Viannia) braziliensis and L. (L.) amazonensis with the aim of investigating the growth kinetics and infectivity of these species in the host cell. Results were expressed as the percentage of macrophages infected measured at 24 hr intervals over six days in RPMI - 1640 culture medium at a temperature of 34-35 degrees C. The findings open the possibility of using canine peritoneal cells as a model for the screening of leishmanicide drugs and to study the pathogenesis of these species. PMID- 10464410 TI - Polymorphism in trypomastigotes of Trypanosoma (Megatrypanum) minasense in the blood of experimentally infected squirrel monkey and marmosets. AB - Experimental infections by Trypanosoma (Megatrypanum) minasense were performed in primates - Saimiri sciureus and Callithrix penicillata - with the objective of searching for morphological variations of the blood trypomastigotes with respect to hosts and time of infection. We carried out morphological and morphometric analysis of blood trypomastigotes. Illustrations are given. Both the squirrel monkey and marmoset became infected after the injection of blood trypomastigotes of T. minasense, although the parasitaemia were briefer in the squirrel monkey. The parasites detected in the later host were narrower and shorter than those found in the inoculated marmoset. In the marmoset, the blood stream parasites derived from culture metacyclic trypomastigotes were considerably smaller than those derived from the inoculation of infected blood. Stronger evidence of polymorphism was found when, at the same time of infection, the blood trypomastigotes found in squirrel monkey had smaller length, body width and the distance from posterior end of the body to the kinetoplast almost four times smaller than the parasite found in the marmoset. Therefore, conflicting results on morphology and morphometry of T. minasense obtained by previous investigators could be due to polymorphism. PMID- 10464411 TI - Genetic polymorphism among six Trypanosoma cruzi stocks isolated from chronic chagasic patients. PMID- 10464412 TI - Absence of Tylodelphys barilochensis (Trematoda: Diplostomidae): induced mortality in "puyenes" Galaxias maculatus (Teleostomi: Galaxiidae) from Southern Argentina. PMID- 10464413 TI - Fatal Plesiomonas shigelloides in a newborn. PMID- 10464414 TI - Effects of chronic chagasic infection on the number and size of cardiac neurons of the wild rodent Calomys callosus. PMID- 10464415 TI - Plasmodium falciparum malaria: rosettes are disrupted by quinine, artemisinin, mefloquine, primaquine, pyrimethamine, chloroquine and proguanil. AB - An assay was developed measuring the disruption of rosettes between Plasmodium falciparuminfected (trophozoites) and uninfected erythrocytes by the antimalarial drugs quinine, artemisinin mefloquine, primaquine, pyrimethamine, chloroquine and proguanil. At 4 hr incubation rosettes were disrupted by all the drugs in a dose dependent manner. Artemisinin and quinine were the most effective anti-malarials at disrupting rosettes at their therapeutic concentrations with South African RSA 14, 15, 17 and The Gambian FCR-3 P. falciparum strains. The least effective drugs were proguanil and chloroquine. A combination of artemisinin and mefloquine was more effective than each drug alone. The combinations of pyrimethamine or primaquine, with quinine disrupted more rosettes than quinine alone. Quinine may be an effective drug in the treatment of severe malaria because the drug efficiently reduces the number of rosettes. PMID- 10464416 TI - Antibacterial activity of Ocimum gratissimum L. essential oil. AB - The essential oil (EO) of Ocimum gratissimum inhibited Staphylococcus aureus at a concentration of 0.75 mg/ml. The minimal inhibitory concentrations (MICs) for Shigella flexineri, Salmonella enteritidis, Escherichia coli, Klebsiella sp., and Proteus mirabilis were at concentrations ranging from 3 to 12 microg/ml. The endpoint was not reached for Pseudomonas aeruginosa (>=24 mg/ml). The MICs of the reference drugs used in this study were similar to those presented in other reports. The minimum bactericidal concentration of EO was within a twofold dilution of the MIC for this organism. The compound that showed antibacterial activity in the EO of O. gratissimum was identified as eugenol and structural findings were further supported by gas chromatography/mass spectra retention time data. The structure was supported by spectroscopic methods. PMID- 10464417 TI - Field studies with the bacterial larvicide INPALBAC for Simulium spp. control in Rio Grande do Sul, Brazil. PMID- 10464418 TI - A new strain of Bacillus thuringiensis serovar israelensis very active against blackfly larvae. PMID- 10464419 TI - Comparison of some behavioral and physiological feeding parameters of Triatoma infestans Klug, 1834 and Mepraia spinolai Porter, 1934, vectors of Chagas disease in Chile. AB - There are two vectors of Chagas disease in Chile: Triatoma infestans and Mepraia spinolai. We studied the feeding behavior of these species, looking for differences which could possibly explain the low impact of the latter species on Chagas disease. Both species used thermal cues to locate their feeding source and consumed a similar volume of blood which was inversely related to the body weight before the meal and directly related to the time between meals. The average time between bites were 6.24 and 10.74 days. The average bite of M. spinolai lasted 9.68 min, significantly shorter than the 19.46 min for T. infestans. Furthermore, while T. infestans always defecated on the host, this behavior was observed in M. spinolai in only one case of 27 (3.7%). The delay between the bites and defecation was very long in M. spinolai and short in T. infestans. These differences may affect the reduced efficiency of transmission of Chagas infection by M. spinolai. PMID- 10464420 TI - Influence of altitude, latitude and season of collection (Bergmann's rule) on the dimensions of Lutzomyia intermedia (Lutz & Neiva, 1912) (Diptera, Psychodidae, Phlebotominae). AB - The influence of altitude and latitude on some structure sizes of Lutzomyia intermedia was noted; several structures of insects collected in higher localities were greater, according to Bergmann's rule. This influence was more remarkable in two localities of the State of Espirito Santo, probably due to greater differences in altitude. Comparing insects from different latitudes, more differences were noted in comparisons of insects from low altitude localities than in those of material from higher altitudes. The small number of differences between insects collected in July and in December does not indicate a defined influence of season and temperature on the size of adults. The possible epidemiological implications of these variations are discussed. PMID- 10464421 TI - Microbial flora variations in the respiratory tract of mice. AB - A stable microbial system in the respiratory tract acts as an important defense mechanism against pathogenic microorganisms. Perturbations in this system may allow pathogens to establish. In an ecological environment such as the respiratory tract, there are many diverse factors that play a role in the establishment of the indigenous flora. In the present work we studied the normal microbial flora of different areas of the respiratory tract of mice and their evolution from the time the mice were born. Our interest was to know which were the dominant groups of microorganisms in each area, which were the first capable of colonizing and which dominated over time to be used as probiotic microorganisms. Our results show that Gram negative facultatively anaerobic bacilli and strict anaerobic microorganisms were the last ones to appear in the bronchia, while aerobic and Gram positive cocci were present in all the areas of the respiratory tract. The number of facultative aerobes and strict anaerobes were similar in the nasal passage, pharynx instilled and trachea, but lower in bronchia. The dominant species were Streptococcus viridans and Staphylococcus saprophyticcus, followed by S. epidermidis, Lactobacilli and S. cohnii I which were present on every studied days but at different proportions. This paper is the first part of a research topic investigating the protective effect of the indigenous flora against pathogens using the mice as an experimental model. PMID- 10464422 TI - Effects of environmental temperature on life tables of Rhodnius neivai Lent, 1953 (Hemiptera: Reduviidae) under experimental conditions. AB - Changes in life tables of Rhodnius neivai due to variations of environmental temperature were studied, based on nine cohorts. Three cohorts were kept at 22 degrees C, three at 27 degrees C and three at 32 degrees C. Cohorts were censused daily during nymphal instars and weekly in adults. Nine complete horizontal life tables were built. A high negative correlation between temperature and age at first laying was registered (r=-0,84). Age at maximum reproduction was significantly lower at 32 degrees C. Average number of eggs/female/week and total eggs/female on its life time were significantly lower at 22 degrees C. Total number of egg by cohort and total number of reproductive weeks were significantly higher at 27 degrees C. At 32 degrees C, generational time was significantly lower. At 27 degrees C net reproductive rate and total reproductive value were significantly higher. At 22 degrees C, intrinsic growth, finite growth and finite birth rates were significantly lower. At 22 degrees C, death instantaneous rate was significantly higher. PMID- 10464423 TI - Ultrastructural features of the astrocytes involved in the phagocytic response to excitotoxins and anoxia in vitro. AB - The present ultrastructural studies demonstrate the morphological evidence of phagocytic response of astroglia to excitotoxins and anoxia in vitro. The studies were performed on organotypic cultures of the rat hippocampus exposed to various excitatory amino acids (QUIN, KA, GLU) or pure nitrogen atmosphere. In all these pathological conditions, the extensive neuronal degeneration was accompanied by marked astroglial reaction. The astrocytes revealed ultrastructural abnormalities consisting of swelling of their cytoplasm followed by more or less advanced fibrillar changes. Numerous reactive astrocytes demonstrated morphological evidence of phagocytic activity during the process of neurodegeneration. They exhibited the presence of ingested degenerating neurons or neuronal debris and accumulation of dense bodies. The results support a possible phagocytic role of astroglia after neuronal injury in various pathological states. The phagocytic response of astrocytes may interfere with other glial functions in CNS and may play a role in tissue recovery. PMID- 10464424 TI - Expression of cyclooxygenase-2 in astrocytes of human brain after global ischemia. AB - COX-2 expression, an inducible form of cyclooxygenase was studied in human brain after ischemia-reperfusion. Previously, a prominent COX-2 upregulation was described in neurons few hours or days after ischemia but little is known about COX-2 expression in non-neuronal cells participating in post-ischemic inflammation. Aim of the study was to examine COX-2 expression in activated glial cells of individuals, who died two or more weeks after resuscitation. In the cerebral cortex of these individuals ischemic necrosis was more or less widespread. In some brain areas numerous microglial cells were found. At the centre of the necrosis high expression of COX-2 was detected in macrophages, polymorphic cells and leukocytes. At the margin of necrosis hypertrophic astrocytes were COX-2 immunopositive. Expression of COX-2 was observed also in the wall of blood vessels of necrotic brain areas and in meninges. The results of the study suggest that in the late period after global ischemia reactive and hypertrophic astrocytes and macrophages may be the major sources of prostaglandins in the human brain. PMID- 10464425 TI - Do astrocytes participate in rat spinal cord myelination? AB - Astrocytes play an important role in CNS development phenomena, such as neuron migration and blood-brain barrier formation, but only a little is known of their role in the process of myelination. The aim of our investigation was to examine the relationship between astrocytes and myelin formation. We evaluated rat spinal cords using hematoxylin-eosin and Kluver-Barrera staining methods as well as immunohistochemical methods with antibodies against myelin basic protein (MBP), glial fibrillary acidic protein (GFAP) and lamins A/C and B2. Our investigation revealed that myelination in the rat spinal cord tracts began between the 6th and 9th postnatal day involving the anterior funiculi, then the lateral funiculi and later the posterior ones. The process of myelination finished about the 25th postnatal day. More GFAP immunoreactive astrocytes were detected in parallel to the increase of MBP reactivity. We suggest that the temporary increase of GFAP positive cells accompanying the process of myelination is necessary for normal myelin development and may be connected with local secretion of growth factors by astrocytes. PMID- 10464426 TI - Secretory meningioma of the brain. Report of two cases. AB - Two cases of cerebral secretory meningioma, occurring in 57 and 33-year-old females are reported. The tumors were located in the tentorial and frontotemporal region, respectively. The general histologic appearance of the tumors was of meningothelial meningioma (case 1) and meningioma with microcystic and angiomatous features (case 2). The most striking histological finding in both tumors were numerous pseudopsammoma bodies, localized chiefly around blood vessels. The inclusions were slightly eosinophilic, stained strongly with PAS method and were differing in size from 3 to 30 microns. Tumor cells containing or surrounding pseudopsammomas were immunopositive for cytokeratin and epithelial membrane antigen. In the first case, individual pseudopsammomas were strongly positive for carcinoembryonic antigen. Some diagnostic aspects of this antigen and problems regarding differential diagnosis in secretory meningioma are briefly discussed. PMID- 10464427 TI - Necrosis and apoptosis of tumor cells in embolized meningiomas: histopathology and P53, BCL-2, CD-68 immunohistochemistry. AB - The preoperative embolization of intracranial meningiomas, used in selected patients to reduce tumor vascularity and blood loss during surgery, may produce ischemic changes and/or tumor necrosis. The aim of the present study was to determine the relationship between necrosis within the embolized tumors and expression of two apoptosis-associated proteins (p53 and bcl-2) and macrophage monocyte CD-68 antigen. Four biopsy specimens of embolized meningiomas, including three benign and one atypical tumor, were revived histopathologically and examined immunohistochemically using the monoclonal antibodies to p53, bcl-2 proteins and CD-68 antigen. The observations showed that the p53-immunopositive cells were most frequent in perinecrotic and ischemic areas than in non-ischemic, intact parts of tumors. The bcl-2 protein was expressed predominantly in well preserved regions lacking ischemic tumor cells, whereas in close proximity to the necrosis only a few bcl-2 positive cells could be detected. Anti-CD-68 immunostained cells were distributed around or within the necrotic foci. Our results indicate that the expression of apoptosis-related proteins correlates with ischemic cell injury induced by preoperative tumor embolization. PMID- 10464428 TI - Quinolinic acid and GABA-B receptor ligand: effect on pyramidal neurons of the CA1 sector of rat's dorsal hippocampus following peripheral administration. AB - In this study we evaluated the effect of baclofen on excitotoxic action of quinolinic acid in hippocampus following its prolonged systemic administration in rats. Male Wistar rats, weighing 200-220 g, were used in the study. Quinolinic acid and baclofen were administered alone or together. Quinolinic acid was administered intraperitoneally (i.p.) in a dose of 60 mmol, baclofen in a dose of 2 mg/kg, by gastric tube, once daily for 8 days. The control group received 1 ml of saline i.p. once daily for 8 days. Quinolinic acid alone produced neurotoxic effect in the CA1 area of the hippocampal formation. The presence of the dark degenerated pyramidal cells was a common sign of a delayed excitotoxic effect. Baclofen added to quinolinic acid markedly attenuated the neurotoxic effect of quinolinic acid. In such cases, only some dark degenerated cells were seen. Baclofen alone resulted in alterations in some pyramidal cells in the hippocampal formation. PMID- 10464429 TI - Cytotoxic activity of serum and cerebrospinal fluid of amyotrophic lateral sclerosis (ALS) patients against acetylcholinesterase. AB - The activity of acetylcholinesterase (AChE) was tested in serum of 20 cases of amyotrophic lateral sclerosis (ALS), 4 "disease controls" and 20 age-matched healthy normals. The AChE activity has been tested also in cerebrospinal fluid (CSF) of 20 ALS patients, 2 "disease controls" and 10 normal subjects. An increase in serum AChE was present in the majority of ALS patients with a mild course of the disease, in the severe ALS group elevated serum AChE activity was a rare finding. Serum ACHE was also increased in multifocal motor neuropathy (MMN). In the majority of mild and severe ALS the CSF AChE activity was decreased. No AChE changes were found in CSF of the "disease controls". Serum and CSF ultrafiltrates of ALS patients and "disease controls" were modifying in vitro the spinal cord AChE activity. In the mild ALS group serum and CSF ultrafiltrates with high molecular weight compounds were decreasing the AChE activity. On the other hand in the severe ALS group serum and CSF ultrafiltrates with low molecular weight compounds were increasing the AChE activity. AChE was modified also in some of the "disease controls", especially in MMN and Guillain-Barre syndrome (GBS) by serum ultrafiltrates containing high molecular weight compounds. The AChE activity in serum and CSF is the consequence of the enzyme leakage from brain, degenerating cholinergic neurons and neuromuscular junctions. We suggest that because of the evoked peripherally divergent changes of the enzyme activity, the AChE values in serum and CSF in ALS do not equal to the degree of the changes in the affected tissues and cannot be taken into account in the prognosis of the disease in particular ALS cases. PMID- 10464430 TI - Diversity of V delta-J delta gene rearrangement in peripheral blood lymphocytes and intrathecal IgG synthesis in multiple sclerosis. AB - The object of the study is a comparison of intrathecal IgG synthesis and gamma/delta TCR genes rearrangement in multiple sclerosis. The subgroup of 13 cases with intrathecal IgG synthesis and positive oligoclonal bands was compared with 8 cases with IgG index below 0.75 and with undetectable oligoclonal bands. TCR gene rearrangement was studied in peripheral blood lymphocytes by PCR analysis. In majority of cases of the first group the V delta-J delta junctional repertoire was restricted as evidenced by oligoclonal rearrangement. Monoclonal pattern of rearrangement was also established in some cases concerning V delta 1 J delta 1 and V delta 5-J delta 1. In all cases with one exception, demonstrating IgG index < 0.75 and with negative oligoclonal bands in CSF the oligo- or polyclonal pattern of V delta-J delta gene rearrangement was noticed. It is therefore suggested that subset T and B lymphocytes may undergo clonal expansion in MS as evidenced by restricted pattern of V delta-J delta rearrangement and intrathecal oligoclonal IgG synthesis, respectively. Oligoclonal expansion at certain B and T cells may occur due to stimulation by an antigen related to MS pathogen. PMID- 10464431 TI - Leprosy: applying qualitative techniques to research and intervention. PMID- 10464432 TI - How Mycobacterium leprae infects peripheral nerves. PMID- 10464433 TI - Nerve function impairment in leprosy: design, methodology, and intake status of a prospective cohort study of 2664 new leprosy cases in Bangladesh (The Bangladesh Acute Nerve Damage Study). AB - The Bangladesh Acute Nerve Damage Study (BANDS) is a prospective cohort study designed to investigate epidemiological, diagnostic, therapeutic and operational aspects of acute nerve function impairment in leprosy. The study is based at a single centre in Bangladesh, in an area with a high prevalence of leprosy. The centre, Danish Bangladesh Leprosy Mission, has a well-established vertical leprosy control programme. In this paper, the study design and methodology are described, together with definitions of nerve function impairment (NFI) used in this and subsequent papers. The study recruited 2664 new leprosy cases in a 12 month period. The male:female ratio is 1.25:1, and 17.61% of the cohort are under 15 years of age. In all, 83.33% of the cohort are paucibacillary (PB), and 16.67% multibacillary (MB). However, the MB rate amongst males is 19.72%, and amongst females is 12.85%, despite an equal period of delay to diagnosis. 55% of patients presented for treatment within 12 months of developing symptoms 6.12% of the total number of cases were smear positive, and 36.71% of the MB cases were smear positive. 9.61% of the total number of cases were graded as having World Health Organisation (WHO) disability grade 1, and 5.97% had grade 2. Amongst MB cases, 27.48% had WHO grade 1 disability present, and 18.24% had grade 2 present, compared with 6.04% and 3.51%, respectively, amongst PB cases. A total of 11.90% of the cohort had sensory NFI of any kind, and 7.39% had motor NFI. Ninety patients presented with NFI needing treatment (3.38%), and of these, 61 (67.78%) had silent NFI. MB patients had a prevalence of reaction/NFI needing treatment nearly 7 times higher than PB cases (15.32% amongst MB; 2.30% amongst PB), and males nearly double that of females (5.67% amongst males, 2.96% amongst females). The most commonly affected nerve by function impairment was the posterior tibial (sensory) with 6.46% of nerves affected (9.38% of patients), followed by the ulnar nerve with 3.23% of nerves impaired (5.56% of patients). Future research and publications, building on this foundation, will focus on the following areas: the incidence of NFI and reactive events, the risk factors for developing NFI, and the response to treatment of patients developing acute NFI. PMID- 10464434 TI - Case detection, gender and disability in leprosy in Bangladesh: a trend analysis. AB - A trend analysis is presented of all newly detected leprosy cases over an 18-year period (1979-1996) in a highly leprosy endemic area of Bangladesh. A total of 23,678 new cases were registered, with an average of 860 new cases per year in the first 12 years, and increasing to around 3000 in 1996. The male:female (M:F) ratio decreased from 2.3 to 1.4. The proportions of newly detected cases with MB leprosy and of newly detected cases with any disability decreased over time. These reductions were more marked in the higher age groups of both sexes. The reduction in disability was primarily attributable to a decline in grade 2 disability. New case detection rates (NCDR) of all leprosy patients per 10,000 general population increased for males from 3 to 6; and for females from 1 to 4, while the NCDR of MB leprosy decreased in males from 1.4 to 0.6, and in females fluctuated around 0.45. The NCDRs of leprosy patients with disabilities showed an initial decrease in the first period, especially in males, but later showed an increase. The NCDR of males with disability was about twice as high as that of females. Finally, female NCDRs in the ages between 15 and 30 were low by comparison with the male NCDRs at the same time. This may be due to the sociocultural characteristics of the Bangladeshi society, with gender differences in exposure, health seeking behaviour and opportunities for case detection. Operational changes in the control programme have contributed to the changed profile of newly detected cases. This study shows that the application of general population statistics is essential for understanding the dynamics in leprosy control programmes under changing operational conditions. Combining case detection figures with such statistics helps to identify population groups that are possibly not benefiting sufficiently from the services provided, and to clarify the dynamics in control programmes and the future trends and programme requirements. PMID- 10464435 TI - Leprosy elimination through integrated basic health services in Myanmar: the role of midwives. AB - Myanmar is one of the top 16 countries identified by WHO as being hyperendemic for leprosy. Multi-drug therapy (MDT) was introduced in 1988 as a vertical programme and gradually integrated into the basic health services (BHS), achieving 100% coverage over the registered cases by 1995. To achieve maximum coverage of and benefit for patients, both leprosy vertical staff and BHS staff were trained to implement MDT whilst performing routine BHS activities. This included a total of 8615 trained midwives who were mobilized for the nationwide leprosy elimination programme (LEP). They worked at village level in various parts of the country and were willing and able to carry out basic tasks in leprosy management, such as the implementation of MDT using blister-calender packs carrying a month's supply of drugs. This study was performed to assess the workload of midwives and their attitude towards LEP. The authors conclude that midwives in Myanmar show a high level of commitment and reliability, which are essential contributing factors to achieve the current goal of leprosy elimination by the year 2000. Along with the present trend of decreasing prevalence rate, leprosy could no longer be considered as a public health problem at national level by the year 2000 in Myanmar. However, because of its long incubation period, new leprosy patients may arise even after the elimination target is achieved, whilst many other patients may become disabled. A community-based sustainable approach for the post-elimination phase, after the year 2000, will be essential and the contribution of the midwives may be of considerable importance. PMID- 10464436 TI - Grading impairment in leprosy. AB - The aim of the paper is to discuss the concept of 'severity grading' in relation to impairment in leprosy, and to describe the use of an impairment sum score, the Eyes, Hands, Feet (EHF) score, as an indicator of the severity and the evolution of impairment over time. The use of an impairment sum score, the EHF score, is illustrated using data on impairment at diagnosis and after a 2-year interval from MB patients released from MDT in the Western Region of Nepal. The WHO 1988 'disability' grading scale (0-2, for both eyes, hands and feet--six sites) was used as a measure of impairment. For the analysis, the WHO grades for the six sites were summed to form an EHF score (minimum 0, maximum 12). The sensitivity to change over time of the EHF score was compared with that of the 'method of maximum grades'. Using the 'method of maximum grades', 509/706 patients (72%) appeared not to have changed in impairment status, compared with only 399 (56.5%) with the EHF score. Improvement or deterioration of impairment status was missed in 113 patients (16%). In 216/706 patients (30.6%), the changes detected with the EHF score were bigger than those revealed by the method of maximum grades. The six components of the WHO impairment grading may be added up to form a EHF sum score of impairment. This score can be used to monitor changes in impairment status in individuals or in groups. It should be recorded and reported at least at diagnosis and release from treatment. Reporting could be done as the 'proportion of patients with improved EHF score', 'stable EHF score' and 'EHF score worse', and 'proportion of patients without impairment', 'proportion with WHO grade 1' and 'proportion with WHO grade 2'. It is recommended that the concepts and terminology of the WHO International Classification of Impairments, Activities and Participation (ICIDH-2) be adopted in the field of leprosy, particularly for the areas of prevention of impairment and disability and rehabilitation. The 'WHO disability grade' should be renamed 'WHO impairment grade'. PMID- 10464437 TI - Factors associated with impairments in new leprosy patients: the AMFES cohort. AB - Data on the importance of the delay between onset of symptoms and registration as a risk factor for impairment are sparse. This study investigates the quantitative relationship between this delay, other risk factors and the impairment status in new leprosy patients. It reports on 592 new leprosy patients enrolled in 1988 1992 in the prospective ALERT MDT Field Evaluation Study in central Ethiopia (AMFES). The influence of the risk factors sex, age, delay, PB/MB classification in relation to BI, and prior dapsone treatment on the impairment status at intake is analysed. Estimates for the delay are based on patient recall. For the risk factors, odds ratios on impairment and on severity of impairment were calculated using both univariate and multivariate logistic regression. The registration delay was 2 years or more for 44% of new patients. The prevalence of impairment (WHO impairment grades 1 and 2 combined) increased continuously from 36% for new patients with a delay of 0-1 year to 81% for new patients with delays of 4 years or more. This prevalence also increased continuously with age; it rose from 26% in children to 80% for the age group 60 and over. In the multivariate regression, the odds ratios for new patients to be impaired were statistically significant for all delay categories (baseline 1-2 years) and age groups (baseline 15-29 years). No statistically significant differences in odds ratios were observed with respect to sex and PB/MB classification in relation to BI. Overall, 31% of new patients presented with WHO impairment grade 1 and 23% with grade 2. The risk on grade 2 also increased with the registration delay amongst the impaired new patients. Relatively few impaired males and relatively few impaired MB patients with a BI value of 3 or higher had grade 2 impairment. Registration delay and age are the main risk factors for presentation with impairment. Reduction of delay in central Ethiopia requires re-thinking of control methodologies. The search for ways to reduce delays in diagnosis and treatment should receive high priority in leprosy research and in leprosy control programmes. PMID- 10464438 TI - Ulcer surgery for non-specialist surgeons. AB - In the majority of cases, plantar ulcers in need of surgical intervention can be treated by very simple procedures. Patients benefit from treatment facilities near to their homes. In the process of integration surgery could be made available to leprosy patients in peripheral health units near their homes by training non-specialist surgeons in peripheral health units in basic surgical procedures, aiming at ulcer healing as well as preventing reoccurrence of ulcers. PMID- 10464439 TI - Improved method of reporting disability grades in POD programmes. PMID- 10464440 TI - Nerve growth factor (NGF) concentrations in cultured human keratinocytes exposed to Mycobacterium leprae cell free extract. PMID- 10464441 TI - The tragedy of deformity in childhood leprosy. PMID- 10464442 TI - AIDS cases soar. PMID- 10464443 TI - Amoebiasis revisited: pathogenesis, diagnosis and management. PMID- 10464444 TI - Proton pump inhibitors. PMID- 10464445 TI - Prevalence of anti-delta antibodies in central India. AB - A total of 238 sera samples from cases of hepatitis, renal failure, thalassaemia, healthy health care workers (HCWs) & asymptomatic HBsAG carriers coming from central India from July 1992 to June 1998, were screened for anti-delta antibodies. Among 238 subjects, 206 were reactive for hepatitis B surface antigen (HBsAg) while 32 were HBsAg non-reactive. The prevalence of anti-delta antibodies was low (1.9%) among 54 patients of acute viral hepatitis (AVH) while it was higher (5.7%) among 52 patients of chronic liver disease (CLD). The anti-delta antibodies positivity among 34 patients with hepatic failure was around 15% and all of them were FHF patients. Among multitransfused subjects such as chronic renal failure (CRF) the prevalence of anti-delta antibodies was low (2.3%). None of the apparently healthy HBsAg reactive HCWs and asymptomatic HBV carriers were reactive for anti-delta antibodies. Similarly anti-delta antibodies could not be detected in HBsAg negative viral hepatitis patients. There is a wide variation in the prevalence of anti-delta antibodies in different parts of India. However, overall prevalence of anti-delta antibodies appears to be lower in the Indian population in comparision to western countries. PMID- 10464446 TI - Detection of Clostridium difficile toxin by an indigenously developed latex agglutination assay. AB - An indigenously developed latex agglutination assay using C. sordelli antitoxin was used to screen 211 stool samples received from hospitalized patients. Of 126 samples from patients receiving single to multiple antibiotics for various ailments, 38 (30%) were positive by the toxin assay, whereas only 6/85 (7%) of samples of patients not receiving antibiotics were also positive. Thus, of 211 samples a total of 44 (20.8%) were positive by our toxin assay, giving titers ranging from 1 in 5 to 1 in 320. The test developed by us is simple, rapid, easy and reliable and can be easily adapted to all microbiology laboratories. PMID- 10464447 TI - Risk factors for duodenal ulcer in north India. AB - OBJECTIVES: The present study was designed to evalutate the association of various risk factors such as smoking, alcohol, NSAIDs, inadequate dietary intake of fibres and consumption of spicy foods with chronic duodenal ulcer using a case control design and to establish the association of Helicobacter pylori and duodenal ulcer using different diagnostic techniques in Indian subjects. MATERIALS: A total of 16 consecutive patients with endoscopically proven duodenal ulcer (DU) constituted the test group while 160 subjects with non-ulcer dyspepsia (NUD) were recruited as controls. METHODS: All subjects were interviewed based on a standard questionnaire and underwent an upper gastrointestinal endsocopy wherein multiple biopsies were taken for rapid urease test (RUT), histology and culture for detection of H. pylori. The serum samples of all the subjects were tested for the presence of antibodies against H. pylori using ELISA. RESULTS: There were significantly greater number of smokers (80%) and alcoholics (58%) in the male population of DU group as compared to the controls (49% smokers & 15% alcoholics). Similar trend was seen in relation to history of chronic exposure to NSAIDs (29% in DU & 11% in NUD) and inadequate intake of fibre in diet (66% and 39% respectively). The overall prevalence of H. pylori was 82.5% in DU and 50.6% in NUD [p < 0.001]. CONCLUSIONS: In North Indian subjects, alcohol consumption, smoking, inadequate intake of fibre in diet and use of NSAIDs are the risk factors associated with doudenal ulcer disease. Further, infection with H. pylori is strongly associated with DU in the North Indian population. PMID- 10464448 TI - Duodenal villous adenoma. AB - Duodenal villous adenomas are extremely rare (1/5th of small bowel tumors). In view of their strategic position and risk of malignancy, they require definitive management in specialized care units. We report a case of a 35 year old female patient with a large duodenal (5 cm) villous adenoma causing intussusception of duodenum (a phenomenon rare because of its anatomic location & fixation). PMID- 10464449 TI - Retained surgical sponge: an unusual cause of malabsorption. AB - Retained surgical sponge is an unpleasant surprise in clinical practice. Intraluminal migration of the retained sponge, though rare, can lead to intestinal obstruction and other complications. We describe two cases of retained surgical sponge, both following gynaecological surgery, presenting several years after surgery with features of subacute intestinal obstruction, malabsorption and several years after surgery with features of subacute intestinal obstruction, malabsorption and sever hypoproteinemia which reverted after surgical removal. PMID- 10464450 TI - Aetiology of chronic diarrhoea in tropical children. AB - AIM: We studied the causes and presentations of chronic diarrhoea in post-weaned children. METHODS: Forty seven children (6 months to 12 years) with diarrhoea of more than 2 weeks duration were recruited and subjected to detailed clinical examination and investigations. RESULTS: Primarily on the basis of history, clinical examination and duodenal biopsy, children could be classified into 5 groups namely tropical enteropathy (46.8%), irritable bowel syndrome (10.6%), giardiasis (14.8%), celiac disease (6.8%) and non specific diarrhoea (21.8%). Children with tropical enteropathy improved with broad spectrum antimicrobial therapy, those with irritable bowel syndrome and non specific chronic diarrhoea with dietary interventions while children with celiac disease required definite withdrawal of gluten from diet. PMID- 10464451 TI - What happens to Helicobacter pylori after vagotomy and drainage? AB - H. pylori is currently identified as the dominant risk factor for chronic duodenal ulcer. The effect of surgery in the form of truncal vagotomy and drainage on the H. pylori status is not well known. Forty three patients with obstructed duodenal ulcer who were positive for H. pylori preoperatively by the urease test on the antral mucosal biopsy specimens were recalled for repeat endoscopy and urease test from the same site at 1 month, 3 months, 6 months and more than 1 year after surgery. The H. pylori positivity declined from 100% preoperatively to 69%, 71%, 73% and 80% at these intervals postoperatively respectively. The fall in H. pylori status after surgery was significant at all intervals. Since surgery for duodenal ulcer in the form of truncal vagotomy and drainage is curative for duodenal ulcer in over 90% of patients whereas H. pylori is suppressed in approximately 20% of patients only, it appears that its effect is independent of H. pylori status. Further studies are required to confirm whether a fall in H. pylori positivity in the antrum is absolute or is due to proximal migration of the organism. PMID- 10464452 TI - Duodenal ulcer perforation: have H2 receptor blockers reduced mortality? AB - Elective surgery for peptic ulcer is becoming rare with the use of more effective medical therapy. However, life threatening complications have not reduced in number. A retrospective study was carried out to compare perforation rates per 10,000 admissions, mortality rates from perforated duodenal ulcers per 10,000 admission and the proportion of patients with perforated duodenal ulcer who died, before and after the introduction of H2 receptor blockers in a large teaching hospital in South India. Perforation rates were not significantly different between the two periods under study. There was a small, but statistically significant (p = 0.047) drop in mortality per 10,000 admissions and a significant drop in proportion of patients with perforated ulcer who died (p = 0.028). Inspite of effective medical therapy, there is a subset of patients with duodenal ulcer who continue to perforate. Efforts should be directed towards identifying this subset and offering them early surgery. Mortality rates have not changed significantly. PMID- 10464453 TI - Rectal duplication cyst. PMID- 10464454 TI - Hydatid cyst of spleen. PMID- 10464455 TI - Solitary juvenile polyp of stomach. PMID- 10464456 TI - [Physicians on call and emergency departments]. PMID- 10464457 TI - [Forensic identification of the dead. The Danish identification preparedness]. PMID- 10464458 TI - [Biomechanics in sports and health sciences]. PMID- 10464459 TI - [Hospitalization and convalescence after hysterectomy. Open or laparoscopic surgery?]. AB - Approximately 6,000 hysterectomies are performed per year in Denmark. The major part of these are performed by laparotomy, although the use of laparoscopically assisted techniques is increasing. Laparoscopic surgery seems to lead to a reduction in the need for postoperative hospitalization and convalescence, which is confirmed in the few available randomised and to our knowledge all retrospective studies. The length of stay seems to be reduced from four to two days if the patient is hysterectomised by the laparoscopically assisted or vaginal techniques. The introduction of endoscopic techniques has caused a change in routines and recommendations, but not for women operated by the classical open procedures. In this article we question the relevance of comparing different types of operations given the fact that the routines and recommendations are different. PMID- 10464460 TI - [Many patients treated in the emergency department could be treated by a general practitioner on call]. AB - Misuse of accident and emergency (A&E) departments is a problem throughout the world. Staffing A&E departments with general practitioners has been reported as successful in a few prospective studies. A medical audit of all A&E charts during one week was carried out. The audit-panel included two orthopaedic surgeons and two general practitioners. The charts were divided in two groups: 1) true A&E cases, 2) inappropriate A&E users, who should have visited the general practitioners' acute consultation instead. There was substantial agreement between audit-panel groups that 57% of attenders could visit the general practitioner instead. Ten percent of attenders received no treatment in the A&E department, but were transferred to other departments within the hospital. It is concluded that two-thirds of the A&E department attenders could have visited the general practitioners instead, for treatment or for referral to the relevant hospital department. PMID- 10464461 TI - [Natural history of abdominal aortic aneurysm with and without concomitant chronic obstructive pulmonary disease]. AB - The relationship between abdominal aortic aneurysms (AAA) and chronical obstructive pulmonary disease (COPD), and in particular the suggested common elastin degradation caused by elastase and smoking was analysed by a cross sectional population mass screening study for AAA, and a prospective cohort study of small AAA. All previous computer-hospital-recorded diagnoses were received concerning 4,404 men invited to screening for AAA. One hundred and forty-one had AAA (4.2%). They were asked for an interview, a clinical examination, and a blood sample. Men with an AAA of 3-5 cm were offered annual control-scans to check for expansion. Of COPD-patients, 7.7% had AAA (crude OR = 2.05), however the adjusted OR was only 1.53 after adjusting for other co-existing diseases (p = 0.13). The mean annual expansion was 2.74 mm per year in COPD patients and 2.72 in non-COPD patients, and 4.7 mm in oral steroid-users compared to 2.6 in non-steroid-users (p < 0.05). S-elastin-peptides (SEP) and P-elastase-alpha1-antitrypsin-complexes (PEAC) were negatively correlated to FEV1 in COPD-patients. However, SEP, beta agonist-treatment, and FEV1 was positively correlated to expansion by multivariate regression analysis, while PEAC and S-alpha1-antitrypsin did not influence expansion, suggesting elastase plays a major role in the pathogenesis of COPD but not in AAA. The high prevalence of AAA among patients with COPD is more likely to be caused by medication and coexisting diseases rather than a common pathway of pathogenesis. PMID- 10464462 TI - [Cryptorchism. Outcome of treatment and referral patterns in an unselected group of patients in a 3-year period]. AB - The age of diagnosis and referral together with the efficacy of HCG treatment were studied retrospectively in 196 unselected cryptorchid patients seen over a period of three years. The median age of diagnosis was 2 years and 7 months, whereas the median age of referral was three years later. During the period of study, guidelines for referral and therapy were published in a nationwide journal and in the local region, and a slight but significant fall in age of referral was seen thereafter. The median age of treatment with HCG, 92 patients, was 6 years and 11 months, and median age of surgery, was 7 years and 7 months. The rate of success with HCG was for bilateral testes 41% and for unilateral testes 21%, giving an overall success rate of 30%. This result is lower than previously reported, which is most likely explained by a higher suprascrotal position of testes before treatment. Guidelines and recommendations for referral and therapy seem to influence the time of referral, which is, however, in this study not in accordance with the consensus of definitive treatment before the age of two years. Early diagnosis is recommended and should be followed by referral to a paediatric department with particular interest and knowledge about cryptorchidism. PMID- 10464463 TI - [Effect of specific physiotherapy on chronic pain, functional level and quality of life in osteoporosis. A prospective randomized single-blind placebo-controlled study]. AB - Patients suffering from osteoporotic vertebral fractures are handicapped by pain and reduced quality of life. Our aim was to investigate the effect of a short training program for osteoporotic patients with regard to pain level, use of analgetics and quality of life. We performed a prospective randomized single blinded placebo-controlled study. The training program included general training of balance and muscle strength and stabilization of the back. The participants were randomised to 10 weeks of ambulatory training. Controls and training participants were tested weekly by registration of pain level and analgetic intake. Questionnaires on daily level of function and quality of life were given at the start and after five and 10 weeks. After three months both groups filled out the questionnaires at home. The training group had a significant reduction in pain score and use of analgetics. The distribution of functional score improved during training. Quality of life score improved significantly throughout the study and after three months. In conclusion, this ambulatory training program is effective for training osteoporotic patients with moderately severe pain and the training should be continued. PMID- 10464464 TI - [Peroneal nerve paresis after long-term bed rest in intensive care patients]. AB - Despite careful nursing and physiotherapy, wasting, myopathy and neuropathy are commonly seen in sedated or comatose intensive care patients undergoing long term bedrest. Four cases of peroneal nerve lesion with drop-foot are described in patients with up to eight weeks of immobilisation due to severe infections complicated with multiple organ failure. No other peripheral nerve compression syndromes were found. These isolated nerve lesions could not be related to the metabolic or cerebral status of the patients. To avoid rotation of the hipjoints with secondary muscle contracture the nursing procedure in the ICU had been changed shortly before the incidents of drop-foot were detected. Along with well known prophylactic anti-compression procedures, sandbags were now placed on the lateral side of the knees. After abandoning this new procedure, peroneal nerve lesions have not been seen. PMID- 10464465 TI - [Hypothenar hammer syndrome]. AB - Hypothenar Hammer Syndrome (HHS) is a rare injury. It usually occurs in the dominant hand of middle-aged men whose occupational or recreational activities require the use of the hypothenar of the hand as a hammer. A case of HHS with Raynaud's phenomenon is presented. Ultrasound duplex and colour scanning revealed an aneurysm with thrombosis of the ulnar artery adjacent to the hook of the hamate. Ligation of the ulnar artery was performed which eliminated the flow in the pseudoaneurysm and prevented further microembolisation to the fingers. PMID- 10464466 TI - [Publications]. PMID- 10464467 TI - [Mortality of cervix cancer can be halved]. PMID- 10464468 TI - [About the strange language of manuscripts]. PMID- 10464469 TI - [On alternative therapy]. PMID- 10464470 TI - Plan and operation of the NHANES II Mortality Study, 1992. AB - OBJECTIVES: The NHANES II Mortality Study is a prospective study of adult participants examined in the second National Health and Nutrition Examination Survey (NHANES II) conducted between 1976 and 1980. It was designed to investigate the association between factors measured at baseline and mortality. The methods used in the study are described and assessed in this report. METHODS: The vital status of NHANES II participants who were 30-75 years of age at their examination was ascertained after 12-16 years. Vital status was assessed by searching the National Death Index and the Social Security Administration Death Master File for deaths occurring in the United States. Causes of death were obtained from the National Center for Health Statistics Multiple Cause of Death file or death certificates. To assess how well mortality was ascertained, the survival of the cohort after 5 and 10 years was compared to that of the U.S. population during the same period. RESULTS: As of December 31, 1992, 23.2 percent of the 9,250 cohort members were found to be deceased. The remaining 76.8 percent that were not found to be deceased may be assumed to be alive for analytic purposes. Cumulative survival probabilities for the cohort were generally higher than probabilities calculated from U.S. life table data. Although some differences were expected, these data suggest that after 10 years of follow-up using passive methods, mortality may have been under ascertained for the cohort. Discrepancies between the survival patterns for NHANES II cohort and U.S. vital statistics were larger for black than for white participants, indicating that ascertainment of mortality was poorer among black participants. Researchers using the NHANES II Mortality Study, 1992 data should be aware of these study limitations. PMID- 10464471 TI - Use of dietary supplements in the United States, 1988-94. AB - OBJECTIVES: This report presents estimates of the prevalence of use of dietary supplements among the U.S. population by various demographic and descriptive characteristics, the number of products taken, and types of supplements taken by broad product-type categories. METHODS: The third National Health and Nutrition Examination Survey (NHANES III) is a nationally representative survey of the civilian, noninstitutionalized U.S. population, 2 months of age or over. Participants were asked about their use of vitamin and/or mineral supplements in the past month. Many also reported use of other dietary supplements. RESULTS: Approximately 40 percent of the population took dietary supplements during the month prior to the interview. Females (44 percent) were more likely to take a supplement than males (35 percent). Non-Hispanic white persons (43 percent) were more likely to take supplements than non-Hispanic black persons (30 percent) and Mexican American persons (29 percent). Children 1-5 years of age were major users of supplements. Among adults 20 years of age and older, there was a trend toward increasing use of dietary supplements with age. Higher levels of education, income, and self-reported health status were all positively related to supplement use. Sixty-seven percent of supplement users took only one supplement, with the majority of them taking a combination vitamin/mineral product (46 percent). CONCLUSIONS: A substantial proportion of the U.S. population takes vitamins, minerals, and/or other dietary supplements. PMID- 10464472 TI - Spatially resolved detection of attomole quantities of organic molecules localized in picoliter vials using time-of-flight secondary ion mass spectrometry. AB - Time-of-flight secondary ion mass spectrometry (TOF-SIMS) has been utilized to detect femtomole and attomole quantities of organic species from within silicon nanovials. By using high-density arrays (10,000 nanovials/cm2) it is possible to chemically characterize diverse sample sets within a single chemical image. Molecular sensitivities, for the compounds investigated, very between 85 attomoles and 25 femtomoles, and typical acquisition times are approximately 100 ms per nanovial. These vials are fabricated using photolithography and KOH etching of Si[001] wafers to create wells, with a pyramidal cross section, ranging in size from 25 to 5625 micron 2. The volume ranges from 30 femtoliters to 100 picoliters, respectively. A drawn glass microinjector and solenoid-driven dispenser are utilized to array picoliter volumes of organic compounds into individual silicon nanovials. Solution concentrations typically range from 1 x 10(-2) to 1 x 10(-4) M allowing femtomole and even attomole quantities of material to be dispensed into each vial. PMID- 10464473 TI - Algorithms for validating chiral properties of insulins. AB - The combination of chiral ligand exchange on Cu(II) complexes in aqueous base with circular dichroism spectropolarimetric detection provides excellent avenues to validate the chirality properties of oligopeptides and proteins. The method is quick and simple and has the potential for development into an automated, routine procedure for quality control applications. Target analytes used for this first study of a protein system are human, porcine, and bovine insulins prepared by different procedures and obtained from different sources, production lots, and manufacturers. The analytical specificity of the test makes the method a potentially useful technique for validating the chirality properties of many peptide and protein forms. PMID- 10464474 TI - Toward a simple, expedient, and complete analysis of human hemoglobin by MALDI TOFMS. AB - MALDI mass spectrometry is explored as a method for hemoglobin characterization. To simplify and expedite the analysis, hemoglobin is obtained without purification directly from whole human blood. The use of trypsinactivated bioreactive MALDI probes is evaluated as a means to further reduce the analysis time from hours to minutes. Moreover, variations of the MALDI matrix preparation facilitate detection of the problematic tryptic peptides alpha T12, alpha T13, and beta T12. The results reveal that MALDI-based methods are easily implemented, are rapid, and allow detection of traditionally elusive tryptic peptides. PMID- 10464475 TI - Direct analysis and identification of Helicobacter and Campylobacter species by MALDI-TOF mass spectrometry. AB - Campylobacter jejuni, Campylobacter fetus, and Campylobacter coli were compared with Helicobacter pylori and Helicobacter mustelae by direct analysis of individual cultured colonies in 50% methanol-water with a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS). H. pylori and Campylobacter species from blood agar culture produced unique, complex spectra with over 25 different ions in mass/charge (m/z) range from 2,000 to 62,000. A biomarker for H. pylori was centered around m/z 58,268, and H. mustelae was distinguished from H. pylori by its ions at m/z 49,608 and 57,231. Campylobacters could be distinguished from Helicobacters by their lack of ions around m/z 58,000 and 61,000 as well as distinguishing biomarkers of lower m/z: 10,074 and 25,478 for C. coli; m/z 10,285 and 12,901 for C. jejuni; m/z 10,726 and 11,289 for C. fetus. MALDI-TOF MS is a rapid and direct method for detection of these potentially pathogenic bacteria from culture. PMID- 10464476 TI - Multiplexed screening of neutral mass-tagged RNA targets against ligand libraries with electrospray ionization FTICR MS: a paradigm for high-throughput affinity screening. AB - We demonstrate that binding of mixtures of aminoglycosides can be measured simultaneously against multiple RNA targets of identical length and similar (or identical) molecular weight. Addition of a neutral mass tag to one of the RNA targets shifts the detected peaks to a higher mass/charge ratio, where complexes with small molecules can be identified unambiguously. An appropriately placed neutral mass tag does not alter RNA--ligand binding. The utility of this strategy is demonstrated with model RNAs corresponding to the decoding region of the prokaryotic and eukaryotic rRNAs and a mixture of five aminoglycosides. Complexes are observed between the aminoglycoside library and the prokaryotic rRNA model, while no aminoglycoside was observed to bind to the mass-tagged eukaryotic rRNA model. The differential binding data is consistent with the eukaryotic A-site rRNA having a different conformation compared with the prokaryotic A-site that prevents entry and binding of neomycin-class aminoglycosides. Mass spectrometric analysis of neutral mass-tagged macromolecular targets represents a new high throughput screening paradigm in which the interaction of multiple targets against a collection of small molecules can be evaluated in parallel. PMID- 10464477 TI - Phosphopeptide isomer separation using capillary zone electrophoresis for the study of protein kinases and phosphatases. AB - Methods for the rapid separation of phosphopeptide isomers (peptides with the same sequence but with phosphates on different residues) were developed using capillary zone electrophoresis with ultraviolet (CZE-UV) detection. Uncoated, cationic and neutral capillaries were used with both acidic and basic peptides. These methods enabled the assay of several protein kinases (mitogen activated protein kinase, protein kinase A, GST-tyrosine kinase) and phosphatases (acid, alkaline, and protein tyrosine phosphatase) and the determination of the sites of phosphorylation and dephosphorylation. Incubations of nonphosphorylated or phosphorylated peptide with kinases or phosphatases took place directly in the instrument's autosampler and were monitored over several hours using CZE-UV. PMID- 10464478 TI - Comparative study of hydrocarbon, fluorocarbon, and aromatic bonded RP-HPLC stationary phases by linear solvation energy relationships. AB - The retention properties of eight alkyl, aromatic, and fluorinated reversed-phase high-performance liquid chromatography bonded phases were characterized through the use of linear solvation energy relationships (LSERs). The stationary phases were investigated in a series of methanol/water mobile phases. LSER results show that solute molecular size and hydrogen bond acceptor basicity under all conditions are the two dominant retention controlling factors and that these two factors are linearly correlated when either different stationary phases at a fixed mobile-phase composition or different mobile-phase compositions at a fixed stationary phase are considered. The large variation in the dependence of retention on solute molecular volume as only the stationary phase is changed indicates that the dispersive interactions between nonpolar solutes and the stationary phase are quite significant relative to the energy of the mobile-phase cavity formation process. PCA results indicate that one PCA factor is required to explain the data when stationary phases of the same chemical nature (alkyl, aromatic, and fluoroalkyl phases) are individually considered. However, three PCA factors are not quite sufficient to explain the whole data set for the three classes of stationary phases. Despite this, the average standard deviation obtained by the use of these principal component factors are significantly smaller than the average standard deviation obtained by the LSER approach. In addition, selectivities predicted through the LSER equation are not in complete agreement with experimental results. These results show that the LSER model does not properly account for all molecular interactions involved in RP-HPLC. The failure could reside in the V2 solute parameter used to account for both dispersive and cohesive interactions since "shape selectivity" predictions for a pair of structural isomers are very bad. PMID- 10464479 TI - HPLC-accelerator MS measurement of atrazine metabolites in human urine after dermal exposure. AB - Metabolites of atrazine were measured in human urine after dermal exposure using HPLC to separate and identify metabolites and accelerator mass spectrometry (AMS) to quantify them. Ring-labeled [14C]atrazine was applied for 24 h with a dermal patch to human volunteers at low (0.167 mg, 6.45 muCi) and high (1.98 mg, 24.7 muCi) doses. Urine was collected for 7 days. The urine was centrifuged to remove solids, and the supernatant was measured by liquid scintillation counting prior to injection on the HPLC to ensure that < 0.17 Bq (4.5 pCi) was injected on the column. A reversed-phase gradient of 0.1% acetic acid in water and 0.1% acetic acid in acetonitrile became less polar with increasing time and separated the parent compound and major atrazine metabolites over 31 min on an octadecylsilane column. Peaks were identified by coelution with known standards. Elution fractions were collected in 1-min increments; half of each fraction was analyzed by AMS to obtain limits of quantitation of 14 amol. Mercapturate metabolites of atrazine and dealkylated atrazine dominated the early metabolic time points, accounting for approximately 90% of the 14C in the urine. No parent compound was detected. The excreted atrazine metabolites became more polar with increasing time, and an unidentified polar metabolite that was present in all samples became as prevalent as any of the known ring metabolites several days after the dose was delivered. Knowledge of metabolite dynamics is crucial to developing useful assays for monitoring atrazine exposure in agricultural workers. PMID- 10464480 TI - Isotope dilution high-performance liquid chromatography/tandem mass spectrometry method for quantifying urinary metabolites of atrazine, malathion, and 2,4 dichlorophenoxyacetic acid. AB - We have developed an isotope dilution high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) method for quantifying the urinary metabolites of the pesticides atrazine, malathion, and 2,4 dichlorophenoxyacetic acid (2,4-D). Urine samples are extracted with an organic solvent, and the organic fraction is concentrated. The concentrate is then analyzed using HPLC/MS/MS. The limits of detection for the metabolites are less than 0.5 microgram/L (parts per billion) in 10 mL of urine, with a high degree of accuracy and precision. PMID- 10464481 TI - Development of a solid-phase microextraction GC-NPD procedure for the determination of free volatile amines in wastewater and sewage-polluted waters. AB - An analytical procedure for the determination of free volatile C1-C6 amines in aqueous matrixes has been developed and applied to their determination in waste water, primary and secondary effluents, and sewage-polluted river samples. The developed analytical procedure involves headspace sampling using solid-phase microextraction with a poly(dimethylpolysiloxane) coating (100 microns) followed by GC-NPD determination and GC/MS confirmation using a tailor-made PoraPLOT amines capillary GC column for volatile amines. Procedural detection limits were compound dependent but ranged from 3 to 56 micrograms L-1, being close to or lower than the odor threshold concentration, and the reproducibility was ca. 15% (N = 5) in real water samples. The developed analytical procedure is solvent free, cost-effective (no cryogenic trap needed), and faster than existing methods because no derivatization step is involved in the determination. Linearity was compound dependent but ranged at least from 50 to 600 micrograms L-1. PMID- 10464482 TI - Immunoassay techniques for detection of the herbicide simazine based on use of oppositely charged water-soluble polyelectrolytes. AB - Linear water-soluble polyelectrolytes, i.e., poly(methacrylate) polyanion and poly(N-ethyl-4-vinylpyridinium) polycation, were used as carriers for the reactants in immunoassay. The strength of ionic forces through distance and the cooperative binding of oppositely charged chains, the carriers interact with each other at an extremely high rate and affinity. These properties of the polyelectrolytes made it possible to carry out the immunochemical steps of the assay in true solution and then to quickly separate the resulting products from the reaction mixtures. The above approach was applied to an assay for the herbicide simazine. Both enzyme-linked immunosorbent assay (ELISA) and dot blot formats of the immunoassay were evaluated. In the ELISA format, the polycation was adsorbed on the surface of a microtiter plate. A tracer antigen (simazine) was allowed to interact in solution with components of the reaction mixture containing simazine-peroxidase conjugate, specific antibodies, and staphylococcal protein A conjugated with the polyanion, and then the mixture was added to the immobilized polycation. Quick separation of the immunoreactants was achieved due to formation of interpolyelectrolyte complexes between polycation and polyanion molecules. After washing, the microplate wells were filled with a solution of substrate, and the optical density of the reaction products was measured. In the second format, a solution of the same reaction mixture (after incubation) was filtered through a porous membrane, with the polycation adsorbed. The subsequent addition of substrate led to the development of colored spots. Sensitivity of the dot blot format was close to that of the traditional ELISA format using the same reactants, i.e., 0.5 ng/mL. However, the assay was much faster (assay time decreased from 100-120 to 45 min). Sensitivities of the dot immunoassay were 1 ng/mL for densitometric detection and 10 ng/mL for visual detection with a duration of 20 min. The techniques developed here were used for simazine determination in water, milk, and juices. PMID- 10464483 TI - Polypyrrole-entrapped quinohemoprotein alcohol dehydrogenase. Evidence for direct electron transfer via conducting-polymer chains. AB - It is reported for the first time that direct electron-transfer processes between a polypyrrole (PPY) entrapped quinohemoprotein alcohol dehydrogenase from Gluconobacter sp. 33 (QH-ADH) and a platinum electrode take place via the conducting-polymer network. The cooperative action of the enzyme-integrated prosthetic groups--pyrroloquinoline-quinone and hemes--is assumed to allow this electron-transfer pathway from the enzyme's active site to the conducting-polymer backbone. A hypothetical model of the electron transfer is proposed which is supported by the influence of various parameters, such as, e.g., ionic strength and nature of the buffer salts. This unusual electron-transfer pathway leads to an accentuated increase of the K M app value (102 mM) and hence to a significantly increased linear detection range of an ethanol sensor based on this enzyme. PMID- 10464484 TI - A simple cleanup method for the isolation of nitrate from natural water samples for O isotope analysis. AB - The analysis of O-isotopic composition of nitrate has many potential applications in studies of environmental processes. O-isotope nitrate analysis requires samples free of other oxygen-containing compounds. More than 100% of non-NO3- oxygen relative to NO3- oxygen can still be found in forest soil water samples after cleanup if improper cleanup strategies, e.g., adsorption onto activated carbon, are used. Such non-NO3- oxygen compounds will bias O-isotopic data. Therefore, an efficient cleanup method was developed to isolate nitrate from natural water samples. In a multistep cleanup procedure using adsorption onto water-insoluble poly(vinylpyrrolidone), removal of almost all other oxygen containing compounds, such as fulvic acids, and isolation of nitrate was achieved. The method supplied samples free of non-NO3- oxygen which can be directly combusted to CO2 for subsequent O-isotope analysis. PMID- 10464486 TI - Sample stacking during membrane-mediated loading in automated DNA sequencing. AB - Microporous membrane-mediated loading is a novel and efficient sample injection technique for ultrathin slab gel electrophoresis-based automated DNA sequence analysis. The sequencing reaction mixture is spotted directly onto the tabs of the membrane loader, which is then inserted to close proximity of the straight edge of the separation gel. The use of a higher viscosity (> 60 cSt), low ionic strength (conductivity < 320 microS) well solution with a pH of more than one unit lower that of the separation and gel buffer system makes possible efficient stacking of the DNA sequencing fragments at the interface of the gel matrix. When the injection/separation process is initiated by the application of the electric field, a high local electric potential drop is formed across the low ionic strength well solution zone. This results in very fast migration of the DNA sequencing fragments toward the interface of the separation gel, where they stack up against the higher conductivity sieving matrix. During this stacking process, primarily the local pH mediates the actual mobility of the buffer co-ions (borate), forming the leading and terminating zones in the well solution and separation gel, respectively. PMID- 10464485 TI - On-line capillary electrophoresis/microelectrospray ionization-tandem mass spectrometry using an ion trap storage/time-of-flight mass spectrometer with SWIFT technology. AB - The development of a system capable of the speed required for on-line capillary electrophoresis-tandem mass spectrometry (CE-MS/MS) of tryptic digests is described. The ion trap storage/reflectron time-of-flight (IT/reTOF) mass spectrometer is used as a nonscanning detector for rapid CE separation, where the peptides are ionized on-line using electrospray ionization (ESI). The ESI produced ions are stored in the ion trap and dc pulse injected into the reTOF-MS at a rate sufficient to maintain the separation achieved by CE. Using methodology generated by software and hardware developed in our lab, we can produce SWIFT (Stored Waveform Inverse Fourier Transform) ion isolation and TICKLE activation/fragmentation voltage waveforms to generate MS/MS at a rate as high as 10 Hz so that the MS/MS spectra can be optimized on even a 1-2 s eluting peak. In CE separations performed on tryptic digests of dogfish myelin basic protein (MBP) where eluting peaks 4-8 s wide are observed, it is demonstrated that an acquisition rate of 4 Hz provides > 20 spectra/peak and is more than sufficient to provide optimized MS/MS spectra of each of the eluting peaks in the electropherogram. The detailed structural analysis of dogfish MBP including several posttranslational modifications using CE-MS and CE-MS/MS is demonstrated using this method with < 10 fmol of material consumed. PMID- 10464487 TI - Ultrasonic flexural-plate-wave sensor for detecting the concentration of settling E. coli W3110 cells. AB - The flexural-plate-wave (FPW) sensor, a type of ultrasonic sensor, can detect changes in E. coli W3110 concentration in solution as the cells settle onto the sensor under the influence of gravity. A model of the sensor's response to cell settling has been developed and is in good agreement with the experimental data. The FPW technique improves on conventional methods for determining cell concentrations; this technique allows for on-line data collection, is nondestructive, and requires only small sample volumes. The FPW sensor has applications as a device to measure cell concentrations and growth rates in industrial fermentors, biofilms, and wastewater treatment facilities. PMID- 10464488 TI - [The ultrasonic measurement of endothelial function]. PMID- 10464489 TI - [The ultrasonic measurement of the endothelial function of the brachial artery in suspected coronary heart disease]. AB - BACKGROUND AND OBJECTIVE: A positive correlation between the presence of coronary artery disease (CAD) and peripheral endothelial dysfunction (ED) of the brachial artery has been shown in several studies. Aim of the present study was to evaluate whether the non-invasive determination of ED could also be used as a screening test in patients suspected of having CAD. PATIENTS AND METHODS: 122 patients were included. 112 had an exercise-ECG before hospital admission. Preceding coronary angiography, FMD% was measured by high-resolution ultrasound (13 Mhz). Longitudinal scans of the brachial artery were done at rest, during reactive hyperemia and after the sublingual administration of nitroglycerin. RESULTS: In 101 of the 122 patients the presence of CAD was diagnosed by angiography, whereas 21 patients had normal coronary arteries. The extent of the vasodilation (FMD%) was found to be largely independent of the resting vessel diameters (FMD%/vessel diameter at rest: r = -0.32767 p = 0.0002). FMD% was significantly higher in patient without CAD than in the CAD group (7.01 +/- 3.5% vs. 3.73 +/- 4.11%, p < 0.001). Comparison of sensitivity and specificity to predict the presence of CAD between FMD% [sensitivity 71.3%, specificity 81%] and exercise-ECG [sensitivity 82.4%, specificity 57.1%] gave similar results. No correlation was found between the degree of the impairment of FMD% and the severely of CAD. CONCLUSION: The determination of peripheral ED was found to be a sensitive and specific measure for predicting the presence of CAD in our cohort. Since this approach is non-invasive, non-radioactive and cost-effective it warrants further evaluation of its role as an additional screening test in patients clinically suspected of having CAD. PMID- 10464490 TI - [The solitary echinococcal cyst--a rare cause of a round pulmonary focus]. AB - HISTORY AND ADMISSION FINDINGS: A 29-year-old man from Kazakhstan presented with right-sided chest pain, cough, whitish sputum and a temperature up to 38.5 degrees C. He had been working in an animal farm for the previous two years. INVESTIGATIONS: Laboratory tests showed increased inflammatory parameters (WBC 15.9 gpt/l with an increased proportion of band and segmented granulocytes, C reactive protein (242.3 mg/dl). Chest radiogram showed an infiltration and effusion in the right base judged to be due to pneumonia. In addition there was a round homogeneous mass, 2.5 cm in diameter, at the lateral thoracic wall in the 8th lower lobe segment. Computed tomography (CT) of the lung and mediastinum confirmed these findings. Bronchoscopy, echocardiography, upper abdominal sonography and CT of the skull revealed no other space-occupying lesion. DIAGNOSIS, TREATMENT AND COURSE: The right-sided pneumonia was successfully treated, at first with ceftriaxone (i.v.), later with ofloxacin (by mouth). An echinococcal cyst was considered especially because of the patient's profession in Kazakhstan in the differential diagnosis of the right-sided round mass and confirmed by an indirect haemagglutination test giving an antibody titre of 1:1024. In response a thoracotomy was performed and the mass, histologically an echinococcal cyst (E. granulosa), resected. There were no other cysts. There was no explanation for the right-sided pneumonia. CONCLUSIONS: Even though Echinococcosis is rare in Central Europe, it should be included in the differential diagnosis of a round pulmonary mass. A detailed history, especially with regard to occupation and country of origin, is essential. Serological tests for specific Echinococcus antibodies, together with usual imaging procedures, will usually and quickly provide the diagnosis. PMID- 10464491 TI - [Morphine poisoning in chronic kidney failure. Morphine-6-glucuronide as a pharmacologically active morphine metabolite]. AB - HISTORY AND ADMISSION FINDINGS: A 57-year-old woman with metastasizing ovarian cancer and chronic renal failure was admitted for morphine treatment of an acute lumbospinal pain syndrome, ambulant treatment with analgesics having failed provide adequate pain relief. On admission due to pain the conscious patient presented with reduced general condition and lumbal pain sensitive to tapping. Lasegue's sign was positive on both sides, no other disturbed neurological functions were found. TREATMENT AND COURSE: On the 7th day of morphine administration she became somnolent and breathing became markedly depressed, indicating overdosage, metabolic and intracranial causes having been excluded. Naloxone, an opioid antagonist, was given i.v. and the breathing pattern improved. But drowsiness continued for another 48 hours and only regressed after repeated doses of naloxone. CONCLUSIONS: Morphine-6-glucuronide (M6G), formed from morphine in the liver, accumulates in blood and penetrates the blood-brain barrier, binding with strong affinity to opiate receptors and exerts a strong analgesic effect. As M6G is excreted by the kidney, its concentration rises in renal failure and can lead to severe intoxication. Morphine dosage must therefore be carefully controlled in patients with renal failure. PMID- 10464492 TI - [The pathogenesis and diagnosis of Rasmussen encephalitis]. PMID- 10464493 TI - [Overweight as a risk factor for cardiovascular diseases and its possible significance as a promotor of an increased inflammatory reaction]. PMID- 10464494 TI - [The AWMF model for the evaluation of published research papers in medicine. Arbeitsgemeinschaft der wissenschaftlichen medizinischen Fachgesellschaften (Working Group of the Scientific Medical Specialty Societies)]. PMID- 10464495 TI - [Vertebral destruction with massive pain in the SAPHO syndrome]. PMID- 10464496 TI - [Vertebral destruction with massive pain in the SAPHO syndrome]. PMID- 10464497 TI - [Standards in digital imaging communication and archiving]. PMID- 10464498 TI - [Magnetic resonance tomography in stroke--its methodological bases and clinical use]. AB - Stroke is the third leading cause of death. Until recently, no diagnostic technique was available that could reliably depict ischemic tissue within a therapeutically promising time window. Since the beginning of this decade, a fundamental change has taken place: Perfusion- and diffusion-MRI have shown to be capable of depicting the ischemic injury immediately after vessel occlusion. While perfusion MRI is sensitive to abnormalities in cerebral capillary blood flow (micro-circulation), diffusion MRI indicates tissue damage on a cellular level. Combining perfusion- and diffusion-MRI with standard MR technique (T2 weighted sequences, MR angiography) results in an MR protocol that is complementary and gives insight into both structural and functional parameters. PMID- 10464499 TI - [Spiral CT of the head-neck area: the advantages of the early arterial phase in the detection of squamous-cell carcinomas]. AB - PURPOSE: To determine if scanning in the arterial phase improves detection of squamous cell carcinomas in the pharynx and larynx. METHODS: In a prospective clinical study 20 patients with a pharyngeal or laryngeal carcinoma were examined with by spiral CT. 80 ml lopromid were intravenously injected as a bolus with a rate of 3 ml/sec. Two consecutive spiral CT scans were performed with start-delay times of 20 and 70 seconds respectively. Delineation and contrast enhancement of tumours, cervical lymph nodes and vessels were evaluated. The radiodensities (HU) of tumors, lymph nodes vessels, pharyngeal wall and muscle were measured. RESULTS: Comparing early and late start delay time scans tumor assessment in the early phase was better in 58%, less in 16% and equal in both scans in 26%. 82% of the pathologic lymph nodes had more peripheral enhancement than surrounding muscle tissue. During the arterial phase the measured radiodensities of the common carotid artery and jugular vein were significantly higher than in the second phase. CONCLUSION: Contrast-enhanced special CT permits accurate morphologic assessment (size, infiltration) of pharyngeal and supraglottic laryngeal squamous cell carcinoma, while pathologic lymph nodes already have a sufficient contrast enhancement for the detection. PMID- 10464500 TI - [The nipple in MR mammography--the normal findings and the signs of disease]. AB - PURPOSE: To differentiate between patterns of contrast enhancement in normal and pathologic nipples. MATERIALS AND METHODS: The examinations were performed on a 1.0 T imager, using a dynamic FLASH-3D-Sequence (9/3/50 degrees). The enhancement of nipples and normal breast parenchyma was evaluated retrospectively in 156 patients. In total, 277 nipples and the parenchyma of 101 patients could be evaluated. In 5 cases histologically confirmed pathologies were found. RESULTS: The 272 normal nipples were characterized by a slow and continuous increase of enhancement from 80% in the first minute to 300% in minute 10. The mean values of the normal parenchyma ranged from 20% in the first minute to 90% in minute 10. The five pathologic nipples showed a high initial signal increase reaching a maximum in the third minute (mean 400%) followed by a plateau. The enhancement of the normal nipples was of fine-linear morphology in the corium, while it was nodular for the pathologic nipples. CONCLUSION: Normal nipples show a slowly increasing fine-linear enhancement, higher than the enhancement of normal parenchyma. Nodular enhancement with a plateau or wash-out should be considered suspicious and a further work-up is necessary. PMID- 10464501 TI - [Acute appendicitis in non-contrast spiral CT: a diagnostic luxury or benefit?]. AB - PURPOSE: To evaluate the diagnostic accuracy of thin collimated unenhanced spiral CT in patients with clinically suspected acute appendicitis and to determine the impact on patient management and overall costs. METHOD: Unenhanced focussed appendiceal spiral-CT was performed in 56 patients (23 women and 33 men) with clinically suspected acute appendicitis. Scans were obtained from the L4 level to the symphysis pubis using 5 mm collimation, 7.5 mm table feed (pitch 1.5) and 4 mm increment without i.v., oral, or rectal contrast material. Prospective diagnoses based on CT findings were compared with surgical (and histopathological) results and clinical follow-up. The effect of spiral-CT on patient management and clinical resources was assessed. RESULTS: 29 patients (10 women and 19 men) underwent appendectomy. Unenhanced spiral-CT was an accurate imaging technique for the initial examination of patients with suspected acute appendicitis with a sensitivity of 95.4% and a specificity 100%, an accuracy of 98.2%, a positive predictive value of 100%, and a negative predictive value of 97.1%. In 27 patients with no evidence of acute appendicitis, an alternative diagnosis could be made in 24 patients by unenhanced spiral-CT. CONCLUSION: Unenhanced spiral-CT is an accurate test to diagnose or to exclude acute appendicitis. Routine appendiceal spiral-CT can improve medical care and reduce the overall costs for patients suspected of having acute appendicitis. PMID- 10464502 TI - [The significance of morphological changes in the brain-tumor border area for the pathogenesis of brain edema in meningiomas: magnetic resonance tomographic and intraoperative studies]. AB - PURPOSE: The aim of the study was to verify a possible correlation between macroscopic changes of the brain-tumor interface (BTI) and the development of a peritumoral brain edema in meningiomas. METHODS: 27 meningiomas were investigated in this prospective study using an optimized inversion-recovery (IR) sequence. After i.v. administration of 0.2 mmol Gd-DTPA/kg axial and coronary images were acquired (slice thickness = 2 mm). The distances of signal altered cortex and obliterations of the subarachnoid space (SAS) were measured at the BTI and related to the pial tumor circumference (cortical-index and SAS-index). Intraoperatively the BTI was divided into the following categories: 0: SAS not obliterated, 1: SAS partially obliterated, 2: direct contact between tumor and white matter, 3: tumor infiltration into brain. RESULTS: Edema-associated meningiomas showed a significantly (p = 0.0001) increased SAS-index (0.47 vs. 0.07) and cortical index (0.45 vs. 0.0) compared to cases without edema. Intraoperatively 95% of meningiomas with brain edema showed SAS-obliterations, compared to 50% of cases without an edema. CONCLUSIONS: Arachnoid adhesions at the BTI with obliteration of the SAS seem to play an essential role in the induction of brain edema in meningiomas. PMID- 10464503 TI - [The simultaneous determination of the arterial input function for dynamic susceptibility-weighted magnetic resonance tomography of the A. carotis interna and the A. cerebri media]. AB - PURPOSE: The determination of the arterial input function (AIF) is necessary for absolute quantification of the regional cerebral blood volume and blood flow using dynamic susceptibility contrast MRI. The suitability of different vessels (ICA--internal carotid artery, MCA--middle cerebral artery) for AIF determination was compared in this study. METHODS: A standard 1.5 T MR system and a simultaneous dual FLASH sequence (TR/TE1/TE2/alpha = 32/15/25/10 degrees) were used to follow a bolus of contrast agent. Slice I was chosen to cut the ICA perpendicularly. Slice II included the MCA. Seventeen data sets from ten subjects were evaluated. RESULTS: The number of AIF-relevant pixels, the area under the AIF and the maximum concentration were all lower when the AIF was determined from the MCA compared to the ICA. Additionally, the mean transit time (MTT) and the time to maximum concentration (TTM) were longer in the MCA, complicating the computerized identification of AIF-relevant pixels. Data from one subject, who was examined five times, demonstrated that the intraindividual variance of the measured parameters was markedly lower than the interpersonal variance. CONCLUSIONS: It appears to be advantageous to measure the AIF in the ICA rather than the MCA. PMID- 10464504 TI - [CT-guided percutaneous biopsies for the classification of focal liver lesions: a comparison between 14 G and 18 G puncture biopsy needles]. AB - PURPOSE: Analysis and comparison of CT-guided 14- and 18-gauge cutting needle biopsies to accurately assess focal liver disease. METHODS: The data of 272 CT guided biopsies in 268 patients were evaluated retrospectively with regard to sensitivity, specificity and complication rate of the chosen needle caliber in differentiating between benign and malignant disease as well as in the ability to determine specific cell types in the various disorders. A 14-gauge (G) needle was used in 101 (37.1%) cases, an 18-g needle biopsy was performed in 171 (62.9%) cases. The Fisher exact test was employed for statistical analysis. RESULTS: Cutting needle biopsy yielded sufficient histologic material in 267 of 272 (98.2%) cases. Correct diagnosis of malignancy was established in 178 of 191 (93.2%) lesions, 73 of 76 (96.1%) disorders were accurately defined as benign, resulting in an overall value of 94.0% (251 of 267). Of these 251 biopsies a definite histological diagnosis could be determined in 90.6% of the cases employing a 14 G needle and in 90.3% using an 18-g needle. The sensitivity, specificity and rate of complications were 94.8%, 92.0%, 3.0% for the 14-g needle and 92.7%, 97.6%, 0.6% for the 18-g needle, respectively. There were no statistically significant differences with regard to the needle size. CONCLUSIONS: CT-guided biopsy of hepatic lesions with the 18-g biopsy needle is of equivalent diagnostic accuracy when compared with the 14-g needle. PMID- 10464505 TI - [A search for the focus in patients with fever of unknown origin: is positron emission tomography with F-18-fluorodeoxyglucose helpful?]. AB - PURPOSE: Search for focus in patients with fever of unknown origin (FUO). MATERIAL AND METHODS: In four patients with the above mentioned problem, F-18-FDG PET was performed, following the common imaging methods, which were without evidence for a focus. RESULTS: The origin of FUO was verified in all patients by PET: Tuberculosis, pneumocystis-carinii pneumonia, chronic inflammatory hematoma, aortitis. CONCLUSION: Successful implementation of F-18-FDG-PET as additional imaging method in patients with FUO seems reasonable. This has to be verified by further prospective studies. PMID- 10464506 TI - [The reduction of the radiation dosage by means of storage phosphor-film radiography compared to a conventional film-screen system with a grid cassette on a skull phantom]. AB - PURPOSE: How much can the radiation dose be reduced for skull radiography by using digital luminescence radiography (DLR) compared to a conventional screen film system with a grid cassette? METHODS AND MATERIALS: A skull phantom (3M) was x-rayed in anterior-posterior orientation using both a conventional screen film system with grid cassette and DLR (ADC-70, Agfa). The tube current time product (mAs) was diminished gradually while keeping the voltage constant. The surface entrance dose was measured by a sensor of Dosimax (Wellhofer). Five investigators evaluated the images by characteristic and critical features, spatial resolution and contrast. RESULTS: The surface entrance dose at 73 kV/22 mAs was 0.432 mGy in conventional screen film system and 0.435 mGy in DLR. The images could be evaluated very well down to an average dose of 71% (0.308 mGy; SD 0.050); sufficient images were obtained down to an average dose of 31% (0.136 mGy; SD 0.065). The resolution of the line pairs were reduced down to 2 levels depending on the investigator. Contrast was assessed as being very good to sufficient. The acceptance of the postprocessed images (MUSICA-software) was individually different and resulted in an improvement of the assessment of bone structures and contrast in higher dose ranges only. CONCLUSION: For the sufficient assessment of a possible fracture/of paranasal sinuses/of measurement of the skull the dose can be reduced to at least 56% (phi 31%; SD 14.9%)/40% (phi 27%; SD 9.3%)/18% (phi 14%; SD 4.4%). Digital radiography allows question-referred exposure parameters with clearly reduced dose, so e.g. for fracture exclusion 73 kV/12.5 mAs and to skull measurement 73 kV/4 mAs. PMID- 10464507 TI - [Image quality and radiation exposure in digital mammography with storage phosphor screens in a magnification technic]. AB - PURPOSE: Comparison of image quality between digital phosphor storage plate mammography in magnification technique and a conventional film screen system regarding the special aspect of radiation exposure. MATERIALS AND METHODS: Radiograms of a RMI-mammography phantom were acquired using a conventional film screen system and two digital storage plate systems. Additionally, the radiograms of one digital system were post-processed emphasizing contrast and included in the comparison. RESULTS: The detectability of details in storage plate mammographies with magnification technique is almost equal to that of film screen mammographies. Thereby, lower radiation exposures were necessary using the digital systems. CONCLUSIONS: Based on these results, storage plate mammography in magnification technique is used in clinical routine at our institution. The correct parameters in image postprocessing are of elementary importance for detail detectability. Future studies must show, whether the lower radiation exposure in digital radiograms of the breast, revealing much higher background noise, will allow the same detail detectability as film screen mammographies. PMID- 10464508 TI - [The quantification of energy-rich phosphates in healthy and damaged heart muscle by SLOOP 31P-MR spectroscopy. Spatial localization with optimal pointspread function]. AB - PURPOSE: A quantitative 31P-MR-spectroscopic technique was used to assess the energy metabolism in healthy and diseased myocardium. METHODS: 31P spectra were acquired on a 1.5 T scanner using a 3D-chemical shift imaging technique. Based on the anatomical information provided by 1H images, SLOOP (Spatial Localization with Optimal Pointspread Function) allows to obtain spectra from defined compartments. With SLOOP a free voxel shape with adaption to anatomic structures, e.g. the myocardium, is possible. Absolute values for phosphocreatine (PCr) and adenosine triphosphate (ATP) were determined using an external standard. RESULTS: 31P-spectra showed only minimal contamination by surrounding tissue. The standard deviation for the determined values of healthy volunteers was low. Compared to healthy volunteers, reduced PCr and ATP concentrations were seen for dilative cardiomyopathies and coronary artery disease and unchanged concentrations were observed for hypertensive heart disease. CONCLUSION: 31P-MR spectroscopy with SLOOP allows a non-invasive, quantitative analysis of cardiac energy metabolism. PMID- 10464509 TI - [Pulsatile tinnitus in contralateral carotid stenosis]. PMID- 10464510 TI - [Multiple brown tumors of the paranasal sinuses in tertiary hyperparathyroidism]. PMID- 10464512 TI - [Acute thromboembolic occlusion of the A. poplitea and of the trifurcation originating from a persistent primitive A. ischiadica]. PMID- 10464511 TI - [An arterial compression syndrome due to the ligamentum arcuatum mediale of the diaphragm]. PMID- 10464513 TI - [The embolization of an aneurysm of the arteria suprarenalis]. PMID- 10464514 TI - [Choriocarcinoma of the kidney in a man]. PMID- 10464515 TI - [Documentation in anesthesia--troublesome or helpful?]. PMID- 10464516 TI - [Importance and perspectives of breath analysis]. AB - Quantitative chemical analysis of volatile constituents in exhaled gas can provide useful insights into biochemical processes in the body. Relations between the chemical composition of human exhaled air and inflammatory processes, states of high oxidative activity, inhalation or ingestion of various noxious substances as well as diseases like ARDS, pneumonia or sleep apnea have been described. Clinical interpretation of these findings, however, remains difficult because definite marker substances for certain diseases could not yet be identified, substance concentrations in the exhaled air change under various conditions, and results vary in a wide range. Because of very low substance concentrations in the exhaled air sophisticated analytical techniques are necessary. Analyses are hampered by high water content and numerous contaminants in the samples. A clear distinction has to be made between substances originating from within the patient and those coming from outside of the body. Despite all these limitations there is no doubt that relations exist between the chemical composition of human exhaled air and clinical parameters. Upcoming new analytical techniques will allow more efficient extraction and preconcentration of substances in minute concentrations. Fast track bedside analyses will shortly become possible with the introduction of miniature gas chromatographic and mass spectrometric equipment. This will open a new area for clinical and basic research. PMID- 10464517 TI - [Informing the patient--but how? Ethics and theory of patient information]. AB - Informed consent is one of the most discussed topics of bioethics. This new emphasis on seeking patient's consent is due to the fact that since the 1960s there has been a considerable change in the ideal of patient-physician relationship. There has been an important shift from a physician-based commitment of promoting well-being to a patient-based right of information arising from individual autonomy instead of a Hippocratic paternalism. Consent requirement is ethically derived from the duty to treat persons as ends, not merely as means. So the physician has a special moral obligation to assure the patient's moral agency by reducing the inequality in information between patient and physician. Therefore a morally valid consent has to be a continuing process rather than a singular event. PMID- 10464518 TI - [Rudolf Eisenmenger's biomotor--predecessor of active compression-decompression cardiopulmonary resuscitation]. AB - Lerman mentioned in 1994 Eisenmenger's Biomotor as a precursor of Active Compression-Decompression Cardiopulmonary Resuscitation (ACD-CPR). We attempted to find additional information. We checked Medline 1966-1998, Quarterly Cumulative Index 1916-1926, Quarterly Cumulative Index Medicus 1927-1950 for publications of Eisenmenger and the secondary literature thereof. Rudolf Eisenmenger (1871-1946) published in 1903 a "Device for Artificial Respiration" consisting of an air-tight thoraco-abdominal shield and a foot-operated bellows for generation of alternating pressure and vacuum on the abdomen and lower thorax. He proposed use of the device for patients in cardiopulmonary arrest caused by drowning or intoxication. The device was patented and in 1904 made commercially available. In 1911 he published a successful resuscitation with his device after one hour of "Vacuum and Pressure Massage of the Abdomen" in a case of attempted suicide by hanging. The foot-operated bellows was replaced by an electromotor (hence "Biomotor") in 1924. Experiments on dogs in cardiac arrest were published in 1929. With the methods available not only "normal" tidal volumes and blood pressure, but also carbon dioxide exhalation and transport of intravenous dye to all parts of the body were shown. In 1939 an eight part series was published, describing use of the device as a respirator in several hospitals. Eisenmenger was the first to propose ACD-CPR, to build a device to perform ACD CPR and to use it successfully in a patient. Furthermore, he was the first to propose the "cardiac pump theory" and the first to recognize the connection of carbon dioxide exhalation, cardiac output and prognosis of cardiac arrest. PMID- 10464519 TI - [Cost analysis concerning MRSA-infection in ICU]. AB - OBJECTIVE: MRSA-infection incidences are still rising, because of unreflected use of antibiotic drugs in man and animals. Although some European countries already have an incidence as high as 30% of MRSA infection in ICU-patients, there is no additional financial support for the treatment of MRSA infected patients. METHODS: We investigated all MRSA infected patients of the operative ICU ward of the department of anaesthesia at the university Ulm within the last three years. We calculated all costs for decontamination and special treatment of the MRSA infection as well as the costs for closing beds, because of MRSA precaution and isolation reasons. RESULTS: The average monthly costs for MRSA infected patients is about 3848 EURO for decontamination and treatment, and another 5560 EURO fix costs. The average monthly ICU duration for MRSA patients was 5.8 days, which means a financial loss of 1622 EURO per "MRSA-patient day". This loss is more than two times the price the social security system pays for an ICU-patient. CONCLUSION: These extra costs should be calculated and additionally paid for patients with MRSA-infection in order to obtain a certain quality standard. By achieving this standard the total economy costs for MRSA infection treatment could be reduced. PMID- 10464520 TI - [Recommendations for medical specialist continuing education towards the DGAI and OGARI. Documentation and quality assurance in anesthesia]. PMID- 10464521 TI - [The use of filters for mechanical ventilation in anesthesiology and intensive care medicine: pro]. PMID- 10464522 TI - [The use of filters for mechanical ventilation in anesthesiology and intensive care medicine: con]. PMID- 10464523 TI - [Symptomatic tachyarrhythmia in patients with a cardiac pacemaker in emergency situations]. AB - Symptomatic tachyarrhythmia in the presence of a cardiac pacemaker is challenging in preclinical emergency situations. This article presents a case report and a diagnostic and therapeutic concept for prehospital treatment of such cases. The usefulness of extracorporeal magnet application which is recommended in the literature and the possibilities of antiarrhythmic therapy will be critically discussed. PMID- 10464524 TI - [Fatal fat embolism from an infected femoral shaft during surgical removal of a hip endoprosthesis]. PMID- 10464525 TI - [Postoperative discomfort]. PMID- 10464526 TI - [Reducing anesthetic morbidity and mortality in developing countries: a priority in obstetrics and pediatric surgery]. PMID- 10464527 TI - [Promoting education in anesthesia and intensive care in developing countries]. PMID- 10464528 TI - [Caffeine skinned fiber tension test: application to the diagnosis of susceptibility to malignant hyperthermia]. AB - OBJECTIVE: To assess the reactivity of sarcoplasmic reticulum to caffeine, using the skinned muscle fibre tension test and to compare it with the reference in vitro contracture test in the diagnosis of malignant hyperthermia (HM) susceptibility. STUDY DESIGN: Laboratory investigation. MATERIAL: Muscle biopsies from 63 patients, including 29 classified as susceptible to MH (MHS) and 34 classified as non-susceptible (MHN) according to criteria of the European and the North American MH groups. METHOD: The reactivity to caffeine and halothane of skinned muscle fibres was compared, according to the type of fibres, with the data of the in vitro contracture test. The type of fibres (type I: oxidative, slow; type II: glycolytic, fast) were determined with strontium dose-response curves. RESULTS: The reactivity to caffeine was significantly lower in the MHS group, for both type I and type II skinned fibres. However, in comparison with the data of the in vitro contracture tests, using the ROC curve analysis, the best sensitivity-specificity compromise was 90%-71% and 74%-84% for type I and type II skinned fibres respectively. CONCLUSION: The skinned muscle fibre tension test cannot be used instead of the in vitro contracture test for the diagnostic of MHS. However, it may strengthen the data of the latter. PMID- 10464529 TI - [Practice of spinal anesthesia in a developing country: usefulness of vascular preloading with a 7.5% hypertonic saline solution]. AB - OBJECTIVE: To assess the efficacy of hypertonic saline for prevention of arterial hypotension in patients undergoing spinal anaesthesia in Niger. STUDY DESIGN: Prospective, randomized, double-blinded study. PATIENTS: Fifty adults undergoing scheduled surgery under spinal anaesthesia, allocated either to a hypertonic saline group (HSG) or a isotonic saline group (ISG). METHODS: Over the 15 min prior to anaesthesia, 100 mL of 7.5% saline were infused in patients of HSG, and 100 mL of 0.9% saline in those of ISG respectively. Spinal anaesthesia was performed at the L3-L4 or L4-L5 interspace using either lidocaine 5%, or bupivacaine 0.5% or a mixture of both supplemented with fentanyl. Arterial pressure (AP) and heart rate (HR) were measured the day before surgery, prior to and after spinal anaesthesia, thereafter every 5 min over 30 min and every 10 min thereafter until completion of surgery. Hypotension (30% decrease of systolic AP control value was treated with 500 mL of Ringer lactate solution and in case of failure with ephedrine (5-30 mg i.v.). An isolated bradycardia (HR < 60 b.min-1) was treated with atropine (0.5-1 mg i.v.). RESULTS: Hypotension occurred in two out of 24 patients of the HSG and eight out of 24 of the ISG (P < 0.05). The mean infused volumes of Ringer lactate solution were 387 +/- 218 mL vs 623 +/- 318 mL respectively (P < 0.05). Ephedrine and/or atropine were not required in HSG, however in 7 out of the 24 patients of the ISG. Adverse clinical effects did not occur. CONCLUSION: Hypertonic saline prevents efficiently the occurrence of hypotension during spinal anaesthesia. Considering its ease of preparation, the lack of adverse effects, in patients not suffering arterial hypertension or congestive heart failure, and low cost, hypertonic saline is well adapted for use in a developing country, if isotonic solutions are not available. PMID- 10464530 TI - [Anesthesia practice by nurse anesthetists in French speaking Sub-Saharan Africa]. AB - OBJECTIVE: To evaluate the practice of anaesthesia in French-speaking subsaharian countries. TYPE OF STUDY: Prospective survey. PERSONS: Two hundred seventeen nurse anaesthetists, from 11 different countries. METHODS: Anonymous questionnaire. RESULTS: One third of nurses were practising anaesthesia since less than five years and 1/3 since more than 10. Only 39 (18%) were working in the country side. Thirty seven (17%) had been trained outside subsaharian Africa (in Cuba 6%, France 5%, Morocco 5% and Germany 1% respectively). Two hundred thirteen (98%) were performing general anaesthesia and 169 (78%) regional anaesthesia. Hundred sixty eight (97%) used spinal anaesthesia, 57 (33%) epidural, 31 (18%) intravenous regional anaesthesia, 24 (14%) axillary block, 17 (10%) caudal block and 10 (6%) peripheral block respectively. For regional techniques, disposable devices were available in 50% of cases. For general anaesthesia, thiopental was administered by 193 (89%) and ketamine by 156 (72%) nurse anaesthetists respectively. In 50% of cases, these drugs were used alone. An ECG was available in 40%, a pulse oximeter in 14% and a capnographe in less than 2% of cases. A ventilator was present in 66% of the places, but used only in 30% of them because of the lack of maintenance and training. CONCLUSIONS: In this study, 50% of nurse anaesthetists were working alone. However, this rate is probably under-estimated, as the questionnaire did not consider anaesthesia practice in the country side. PMID- 10464531 TI - [Evolution of the demography of anesthesia practitioners in French speaking Sub Saharan Africa]. AB - OBJECTIVE: To assess the course of the demography of anaesthesia providers in French-speaking subsaharian countries. TYPE OF STUDY: Retrospective survey. PERSONS: Doctors trained in anaesthesia and nurse anaesthetists registered in West African medical societies. METHODS: Countries, hospitals, anaesthesia manpower, seniority and place of training were analysed. RESULTS: In the 13 French-speaking subsaharian countries including 97.5-M inhabitants, 122 doctors and 868 nurses were registered as anaesthetists in 1998. Mean ratios were one doctor trained in anaesthesia for 799,180 inhabitants and one nurse anaesthetist for 112,327 inhabitants. From 1980 to 1998, these figures increased by a factor 11 for doctors (+1100%) and by a factor two for nurses (+100%). Most doctors were working in the chief town, in both public and private health care institutions. CONCLUSIONS: In all French-speaking subsaharian countries, a major shortage of doctors trained in anaesthesia is existing. PMID- 10464532 TI - [Anesthesia practices in Yaounde (Cameroon)]. AB - This descriptive and prospective study, carried out during a 6-month period at Yaounde (Cameroon), assessed work practices relating to anaesthesia in public and private health care institutions, the incidence and causes of complications, and considers possible means for improving security of the anaesthetized patient. This study included 1,103 patients, aged 12 days to 89 years. Main surgical indications were gynaecology-obstetrics (33.3% of cases) and emergency surgery (28.6% of cases). Anaesthetic practices were characterized by an unsuitable and non systematic use of procedures which are essential parts of a safe anaesthetic. Pre-anaesthetic assessment was carried out in 71.4% of cases, premedication in 78.4% of cases and management in recovery room in 24% of cases only. Patients were anaesthetized by nurses in 78.6% of cases (40% of them had been trained on the-job only). The anaesthesia equipment of operating room was poor in most places, especially with respect to physiological monitors. The rate of complications, 476 in 321 patients (29.1% of cases), was similar to the incidence registered in 1977 (30%). Twenty-four deaths occurred (overall mortality rate of 2.2%). Besides the shortage of equipment, this study underlines the necessity for producing and adhering to guidelines for safe practice of anaesthesia, adapted for developing countries. PMID- 10464533 TI - [Training missions in Cambodia: impact on the anesthesiologists and intensive care workers who carried out these missions]. AB - OBJECTIVES: 1) To analyse the incentives and the significance of the participation of anaesthesiologists in training missions for male nurse anaesthetists in Cambodia; 2) to assess the professional and private impact of these missions on them; 3) to evaluate the level of contribution of hospital administration. STUDY DESIGN: Retrospective survey. PERSONS AND METHOD: Twelve anaesthesiologists, mainly males, mean age 44 +/- 6 years, with some expertise in missions in developing countries complete anonymously a written questionnaire including standardized answers and open questions. RESULTS: The mean reason for participating was the wish to transmit knowledge and expertise. For most of them the mission was an extension of their clinical activity. The reverse effects were a change of their mind on medical practice, on teaching and training, assistance projects for developing countries. For the latter, all considered that the health care institutions in association with Faculties of medicine should be involved in these missions. DISCUSSION: Initially, these missions expressed a need of self sacrifice and appraisal of professional expertise. However the educational goal of the mission, the difference of cultural environment, determined a change in links with other persons. As a result, they wished to share expertise with other colleagues, in modifying the procedures for education and their participation in future assistance projects for development. However these intentions remained without professional impact as these educational actions were not included in a real institutional project. CONCLUSION: Every international assistance project for development build by hospital doctors should be examined not only by staff members of the department, but also by the hospital administration. PMID- 10464534 TI - [Locoregional intravenous anesthesia]. AB - OBJECTIVE: To analyse current data on intravenous regional anaesthesia (IVRA), its benefits and drawbacks. DATA SOURCES: Articles were obtained from a Medline search using the following search terms: 'intravenous regional anaesthesia', alone or combined with 'local anaesthetic agents', 'toxicity'. STUDY SELECTION: Following articles in English and in French have been selected: main articles, original articles, update and review articles, letters to the editor and recent editorials. DATA EXTRACTION: Physiopathological and pharmacological data were extracted for involved mechanisms and means for improving this technique. DATA SYNTHESIS: IVRA is a reliable and efficient technique with a lower cost than general anaesthesia and well adapted for limb surgery in the ambulatory patient. Depending on the site of the surgical field, the pneumatic tourniquet is set either on the arm, forearm or wrist for the upper limb or thigh, calf or ankle for the lower limb. When set in periphery, less local anaesthetic agent is required. A wide tourniquet requires a lower inflation pressure than a double cuff tourniquet. A single cuff is as efficient as a dual cuff if shape, size and inflating pressure are appropriate. The limb occlusion pressure (LOP) is the minimal pressure required to occlude blood flow. It is assessed with either a pulse oximeter or Doppler for determination of the lowest cuff inflating pressure. The cuff is inflated to 50 mmHg above LOP. Oozing in the surgical field can be decreased by the re-exsanguination technique. Currently, lidocaine is the only local anaesthetic released in France for IVRA. Addition of a muscle relaxant, a NSAID or clonidine allows the dose of local anaesthetic agent to be decreased and improves postoperative analgesia. PMID- 10464535 TI - [Suprascapular palsy: a complication of surgical positioning?]. AB - We report a case of persisting suprascapular nerve palsy after surgery under general anaesthesia of short duration in a conventional position. Surgical exploration, eight months later, showed a suprascapular notch narrowed by a hypertrophied and calcified superior transverse ligament. Such a lesion and the practice of volley-ball by the patient are in favour of a pre-existing infraclinical neuropathy. PMID- 10464536 TI - [Peripartum cardiomyopathy. Clinical case series]. AB - We report a retrospective analysis of four cases of peripartum cardiomyopathy admitted, within a 10-year period, to the maternity hospital of Sousse. The age of the patients was over 30 years, two had gemellary pregnancies and two suffered toxaemia. Congestive cardiac failure, mainly of the left ventricle, with acute pulmonary oedema, was the main symptom. Chest X-ray showed a cardiothoracic index above 0.58 and echocardiography a left ventricular telediastolic diameter above 5.7 cm and a shortening ratio of 25% or less. Endomyocardial biopsy for ascertaining the diagnosis was not available. Treatment included bed rest, water and salt restriction, digitalis, a diuretic and an anticoagulant in case antepartum cardiomyopathy, a converting enzyme inhibitor and a diuretic in case of postpartum cardiomyopathy. Three patients recovered totally and in one echocardiography showed a persisting impaired left ventricular function. Five neonates had a favourable outcome and one foetus died in utero. Main problems raised by peripartum cardiomyopathy are discussed. PMID- 10464537 TI - [Chloroquine poisoning with respiratory distress and fatal outcome]. AB - Chloroquine poisoning can cause life threatening cardiovascular disturbances. We report three cases of suicide with chloroquine causing acute respiratory insufficiency from pulmonary oedema with lethal outcome, despite a treatment with diazepam, epinephrine and mechanical ventilation. PMID- 10464538 TI - [Severe intoxication probably from olanzapine (Zyprexa). Beneficial effect of glucagon]. AB - We report the case of a 43-year-old schizophrenic who sustained, after 12 days of treatment including olanzapine (20 mg.day-1), carbamazepine, levomepromazine and alprazolan, a severe shock with bradycardia (HR: 40 b.min-1), circulatory collapse (SAP: 60 mmHg), hypothermia (T: 27 degrees C), coma and disseminated intravascular coagulation. A significant improvement was obtained with high doses of intravenous glucagon, whereas the normalization of central temperature, atropine, adrenaline and volume loading had been inefficient. Olanzapine, alone of associated with other psychotropics, could cause severe circulatory collapse with hypothermia and coma responding to a treatment including glucagon. PMID- 10464539 TI - [Delivery of desflurane by a Tec 6 vaporizer at a concentration superior to that posted on the regulating valve]. AB - We report the case of a defective Tec 6 vaporizer (Datex-Ohmeda) which delivered higher concentrations of desflurane (end-tidal concentration: 3.5 vol%) than indicated by the vaporizer dial setting (2 vol%), at a fresh gas flow of 1 L.min 1. This condition was caused by a defective rotary control valve. Therefore it is essential to administer desflurane only in presence of a vapour analyser. PMID- 10464540 TI - [Postoperative nausea and vomiting after laparoscopic cholecystectomy]. PMID- 10464541 TI - [Utilization of a Tuohy needle for insertion of a spinal needle]. PMID- 10464542 TI - [The laryngeal mask and desflurane]. PMID- 10464543 TI - [Use of the ConstaVac blood reinfusion system for postoperative autotransfusion in knee prosthesis surgery]. PMID- 10464544 TI - [Insertion of peripheral intravascular cannulas in the operating room]. PMID- 10464545 TI - [Principles of tests of hypotheses in statistics]. PMID- 10464546 TI - Neuropsychiatric syndromes in systemic lupus erythematosus: a new look. PMID- 10464547 TI - Dose-loading with hydroxychloroquine is effective in early and active rheumatoid arthritis. PMID- 10464548 TI - Activation of progelatinase B in synovial fluids of patients with rheumatoid arthritis, with reference to stromelysin-1 and tissue inhibitor of matrix metalloproteinase-1. AB - OBJECTIVE: To clarify whether stromelysin-1 and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are involved in the modulation of activation of progelatinase B in the synovial fluid of patients with rheumatoid arthritis (RA). METHODS: Gelatinases in the synovial fluid of patients with RA were analyzed by gelatin zymography. Concentrations of stromelysin-1 and TIMP-1 were measured using a specific sandwich enzyme linked immunosorbent assay. RESULTS: Forty-three rheumatoid synovial fluids containing progelatinase B were examined to clarify whether the enzyme was activated by incubation. Incubation at 37 degrees C caused the conversion of progelatinase B to the active form in 22 of the 43 synovial fluids. The levels of both stromelysin-1 and TIMP-1 were determined for each group and the concentration ratio of stromelysin-1/TIMP-1 in the synovial fluids of each group was highly correlated to the activation of progelatinase B. CONCLUSION: The balance between the concentrations of stromelysin-1 and of TIMP-1 in the synovial fluid appears to determine whether the progelatinase B molecule causes conversion into the active form. PMID- 10464549 TI - Impaired diastolic function in active rheumatoid arthritis. Relationship with disease duration. AB - OBJECTIVE: Using digitized M-mode and Doppler echocardiography, we evaluated left ventricular (LV) function in 54 patients (43 women and 11 men; mean age 50 years) suffering from active rheumatoid arthritis (RA) without obvious cardiovascular disease, and compared them with 54 age- and sex-matched normal subjects. RESULTS: No differences were found in LV end-diastolic diameter, systolic function and parietal thickness between the patients and controls. However, a significant reduction in various indexes of LV diastolic function was found, including E/A (ratio of early to late filling waves of mitral inflow Doppler) and the peak lengthening rate of the LV diameter (an index of LV relaxation evaluated by M mode echocardiography). The former was correlated with patient age and was independent of disease duration, while the latter was more markedly correlated with disease duration than with patient age. CONCLUSION: The relationship between diastolic impairment and disease duration in active RA may open new perspectives in the study of RA-associated cardiovascular disease. PMID- 10464550 TI - Body image of women with rheumatoid arthritis. AB - OBJECTIVE: Physical disabilities generally cause severe disturbances in a patient's body image perception. The aim of the present study was to investigate different aspects of body image, including sexual dissatisfaction, in women with rheumatoid arthritis (RA) in relation to their subjective impression of handicap. METHODS: Forty women with RA were investigated using a series of instruments: Strauss and Appelt's questionnaire for assessing one's body (1), the body perception scale of Paulus (2), and an interview focusing on appearance, worries about health and sickness, and sexual dissatisfaction. In addition, clinical parameters and the subjective extent of morning stiffness were documented, and patients with a high degree of morning stiffness were compared to patients with a low degree of morning stiffness. RESULTS: In contrast to patients with a low degree of morning stiffness, patients with a high degree of morning stiffness worried significantly more about their bodies (p < or = 0.05) and reported significantly more problems in sexuality (p < or = 0.05). CONCLUSION: Our results suggest that morning stiffness plays a very important role in how severely a woman feels herself to be handicapped. Severely handicapped women have to deal with anxieties about health and have sexual problems. Physicians should not shy away from addressing these issues and in severe cases psychological therapy should be initiated. PMID- 10464551 TI - Relationship between serum RANTES levels and radiological progression in rheumatoid arthritis patients treated with methotrexate. AB - OBJECTIVE: The aim of this study was to evaluate the relationship between serum chemokines and the clinical and radiological response to a one-year course of methotrexate (MTX) in patients suffering from rheumatoid arthritis (RA). METHODS: Twenty out-patients suffering from active RA entered a one-year open prospective study on the effects of low dose MTX therapy. Plain radiographs of the hands and feet were taken at study entry and at the end of the follow-up, and were compared for the number of eroded joints. Serum levels of both C-X-C and C-C chemokines were obtained before the initation of MTX and after 6 and 12 months of treatment. RESULTS: The levels of serum RANTES before treatment were significantly higher in RA patients than in the controls and returned to normal levels after one year of treatment. Serum levels of the other chemokines were either in the normal range or undetectable. Twelve patients (60%) did not show any new eroded joints at the end of the follow-up period and were considered as radiological responders (RR). Serum levels of GRO-alpha and RANTES after 6 months of treatment were significantly higher among the patients with radiological progression than in RR patients. CONCLUSIONS: We observed high levels of serum RANTES in a series of RA patients during the active stage of the disease. MTX treatment significantly lowered the serum levels of RANTES, GRO-alpha and MCP-1. High levels of serum RANTES or GRO-alpha after 6 months of MTX treatment seem to be predictive of radiological erosions after one year. PMID- 10464552 TI - The utility of the lupus band test on sun-protected non-lesional skin for the diagnosis of systemic lupus erythematosus. AB - OBJECTIVE: The sensitivity and specificity of the lupus band test was evaluated, using three different criteria, on sun-protected non-lesional skin for the diagnosis of systemic lupus erythematosus (SLE). In addition, the sensitivity and specificity of the lupus band test was compared with those of other laboratory tests used in the diagnosis of SLE. METHODS: Sun-protected non-lesional skin biopsies from 65 patients (F 50; M 15; mean age 41 yrs.) with specific cutaneous manifestations of lupus erythematosus (LE) and from 18 patients with other dermatologic diseases (F 11, M 7; mean age 40 yrs.) were tested using the direct immunofluorescent technique. RESULTS: The sensitivity and specificity of the lupus band test was 10.5% and 97.8% respectively using the strict criterion of the presence of two different immunoreactants. The sensitivity and specificity were 52.6% and 69.5% respectively based on the presence of two different immunoreactants and were 78.9% and 47.8% based on the presence of only one immunoreactant. The highest sensitivity was found for ANA (100%). The specificity of all the laboratory abnormalities was particularly high, varying from 82.8% to 100%, except for ANA antibodies which showed a specificity of 65.2%. CONCLUSIONS: A positive lupus band test on sun-protected non-lesional skin (even if showing the presence of only one immunoreactant at the dermo-epidermal junction) represents a useful and specific criterion for identifying patients with LE. However, this test is not useful in distinguishing between cutaneous lupus patients with systemic involvement and those without systemic involvement. PMID- 10464553 TI - In vitro up-regulation of E-selectin and induction of interleukin-6 in endothelial cells by autoantibodies in Wegener's granulomatosis and microscopic polyangiitis. AB - OBJECTIVE: In patients with Wegener's granulomatosis (WG) or microscopic polyangiitis (MPA) autoantibodies to myeloid granule proteins (ANCA), particularly proteinase 3 (Pr3) and myeloperoxidase (MPO), and to endothelial cells (AECA) are frequently detected. The role of these autoantibodies in the development of vascular injury is incompletely understood. Since the expression of E-selectin and the production of interleukin 6 by endothelial cells is an early step in the sequence of events leading to vascular injury, we examined the capacity of IgG fractions from patients with WG and/or MPA to activate endothelial cells to the expression of E-selectin and the production of IL-6. We related those findings to the presence of ANCA and AECA in the IgG preparations. METHODS: Human umbilical vein endothelial cells (HUVEC) were incubated with immunoglobulin (IgG) preparations from 28 patients (17 positive for anti-Pr3, 10 for anti-MPO, and one for anti-Pr3/MPO) with active vasculitis and from 10 healthy volunteers. The final IgG concentration in the activation assay was 2 mg/ml. TNF alpha (10 ng/ml) and LPS (10 ng/ml) were used as positive controls for HUVEC activation. The extent of HUVEC activation was assessed by the measurement of E-selectin expression by flow cytometry (after 4 hours of incubation) and the production of interleukin 6 by ELISA (after 24 hours). RESULTS: We found that 11 of the 28 ANCA positive IgG samples were capable of activating endothelial cells: six samples induced IL-6 production alone, one sample upregulated E-selectin expression alone, and four samples induced both IL-6 production and E-selectin upregulation. Five of 17 anti-Pr3 positive samples (one of which was also positive for AECA) and 6 of 10 anti-MPO positive samples (all simultaneously positive for AECA) induced endothelial cell activation. AECA positive samples that induced endothelial cell activation (n = 7) had higher AECA titres than samples that did not induce endothelial cell activation (n = 6). CONCLUSION: Our data suggest that the activation of endothelial cells in patients with WG and MPA can be induced by circulating autoantibodies. Both ANCA and AECA can be responsible for this effect. PMID- 10464554 TI - Functional outcome of myositis patients: can a low-dose glucocorticoid regimen achieve good functional results? AB - OBJECTIVE: During the last few years, in an attempt to reduce the side effects of glucocorticoid (GC) therapy, we have been treating polymyositis-dermatomyositis (PM-DM) patients with a lower starting dose of GC than is classically recommended. In order to validate this approach, we performed a functional re evaluation of these PM-DM patients. METHODS: A comprehensive protocol evaluating muscle strength, muscle function, CK levels, persistence of spontaneous activity on electromyography, disability in daily life activities and degree of dependence was applied in 25 non-cancer-associated biopsy-proven PM-DM patients, 15 of whom had been treated with a high-dose regimen (i.e. > 0.5 mg prednisolone/kg/day) and 10 with a low-dose regimen (i.e. < or = 0.5 mg prednisolone/kg/day). RESULTS: Our results indicate that the functional outcome of PM-DM patients given a low-dose starting regimen of GC does not differ from that observed in patients given higher doses. Interestingly, vertebral fractures were less common in patients treated with lower GC doses. CONCLUSIONS: Although our analysis has certain shortcomings, including the small number of patients investigated and their uncontrolled assignment to a low-dose or a high-dose GC regimen, the results of this retrospective study suggest that a low-dose starting regimen of GC can achieve a good functional outcome in PM-DM patients, with a reduction of treatment-related disability. This approach would be welcome as a step forward should it be validated by a longitudinal study. PMID- 10464555 TI - Vertebral osteomyelitis in northern Spain. Report of 62 cases. AB - OBJECTIVE: The records of 62 patients with clinical and radiographic evidence of vertebral osteomyelitis and positive bacteriological diagnosis, seen between 1979 and 1996, were reviewed in order to gather data on the epidemiology and the clinical pattern displayed by patients with this condition in northern Spain. RESULTS: Staphylococcus aureus (15 cases), Mycobacterium tuberculosis (15 cases) and Brucella melitensis (13 cases) were the microorganisms most frequently found in our patient series. After improvement of the sanitary and hygienic control of food products, the role of Brucella melitensis is decreasing as a causative agent (only 3 cases in the last 6 years). Staphylococcus epidermidis, present in 4 cases (6.6%), should be suspected in elderly patients with previous intravenous cannulations (3 of 4 cases). The most frequent risk factors were alcoholism (7 cases), chronic hepatic disease (7 cases), diabetes (6 cases) and previous surgery (6 cases). Delay in diagnosis was high (the mean number of days between the onset of symptoms and diagnosis was 125). The lumbar region was the most commonly affected site. Neurologic involvement was present in 10 patients on admission (16%). ESR was > 50 mm/hr in a high number of cases. Blood cultures were found to be the most valuable routine test. Plain x-rays were normal in 10 patients (16%); in 6 of them Staphylococcus aureus was the responsible organism. Other imaging modalities showed a high sensitivity. Surgical drainage was necessary in 12 individuals (in 7 due to Mycobacterium tuberculosis). Outcome was good in the majority of cases: only 2 patients with associated endocarditis died. Neurologic sequelae were present in another 3 patients. CONCLUSION: Vertebral osteomyelitis can be caused by a variety of pathogens. Therefore, bacteriological studies are necessary to establish the etiologic diagnosis and determine the specific antimicrobial treatment required. PMID- 10464556 TI - Disease modifying and immunomodulatory effects of high dose 1 alpha (OH) D3 in rheumatoid arthritis patients. AB - OBJECTIVE: Vitamin D analogues such as 1 alpha (OH) D3 (alphacalcidiol) have a possible physiological paracrine effect on cell proliferation and differentiation. Experimentally established possibilities to prevent autoimmune diseases suggest that alphacalcidiol may have therapeutic value as an immunomodulatory agent in patients with rheumatoid arthritis. METHODS: We organized a 3-month open-label trial on 19 patients being treated with standard DMARD therapy for acute RA. They were divided into 2 subgroups, those with highly active RA and those with moderately active RA. Their regular drug regimen was maintained during the trial and oral alphacalcidiol 2 micrograms/day was added. Therapy results were evaluated by ESR, CRP, morning stiffness, the Richie index, and the Lee index. Immunomodulatory effects were investigated by measuring lymphocyte proliferation and apoptosis both in the patients and in vitro in 10 nM alphacalcidiol-supplemented culture medium. RESULTS: After 3 months, high dose oral alphacalcidiol therapy showed a positive effect on disease activity in 89% of the patients (45% or 9 pts. with complete remission and 44% or 8 pts. with a satisfactory effect). Only two patients (11%) showed no improvement, but no new symptoms occurred. No side effects were observed. CONCLUSION: These results suggest that alphacalcidiol is a powerful immunomodulatory agent with fairly low hypercalcemic activity. Clinical improvement was strongly correlated with the immunomodulating potential of this agent. We noticed dual effects on lymphocyte proliferation and apoptosis according to the prior cell activation state. Alphacalcidiol could therefore possibly be used as an adjunct therapy with DMARDs in patients with rheumatoid arthritis. PMID- 10464557 TI - Osteoporosis with lymphoid nodules and hematopoietic marrow hyperplasia. AB - OBJECTIVE: In 1983 Vigorita reported 3 cases of osteoporosis associated with intramedullary lymphoid nodules. We present 8 patients with osteoporosis and lymphoid nodules (LN) in whom we studied the clinical, biological and histological features and the course of the disease. METHODS: Three men (mean age 52 yrs., range 43-68 yrs.) and 5 women (mean age 60 yrs., 49-66 yrs.), 6 of them with osteoporosis with fracture and 2 with osteoporosis on bone densitometry (T score < -2.5 SD) were enrolled in this study. The following parameters were studied: immunobinding with IG determination, phosphorus and calcium levels, PTH, 25 and 1-25 OH D3, osteocalcin, urinary deoxypyridinoline, histomorphometry, tests for autoanti-bodies, HIV, HTLV, EBV and CMV serology. The results were compared with those of 20 patients with osteoporosis but without LN. Five patients underwent a second BMB a mean of 2 years after the first. RESULTS: Five patients had asthenia, 4 had joint pain and 3 had hyperlymphocytosis. Immunologic and virologic investigations were negative in all cases. Bone marrow was hypercellular (59.9 +/- 5.3 vs 40.1 +/- 13%, p: 0.001). At the second BMB, LN were absent but bone marrow was still hypercellular. In all cases, no cause of demineralization was found and osteoporosis progressed rapidly (an average of 3 vertebral compression fractures in three months, with increased resorption (ES 6.5 +/- 1.6 vs 3 +/- 1.2, p: 0.05) with decreased calcification rate (CR 0.62 +/- 0.07 vs 0.79 +/- 0.1, p: 0.04). CONCLUSION: Some interesting questions are raised by this study. Did an undiscovered viral infection cause the asthenia and joint pain via cytokines or PTHrp in our patients, and can activated lymphocytes perhaps modify bone remodeling? PMID- 10464558 TI - No significant effects of sodium aurothiomalate on haem metabolism and mixed function oxygenase activity in patients with rheumatoid arthritis. AB - OBJECTIVE: Animal studies suggest that gold compounds impair haem synthesis and increase haem degradation and, as a result, reduce activity of the hepatic haemoproteins cytochromes P-450. The aim of this study was to investigate whether intramuscular gold exerts similar effects in patients with rheumatoid arthritis (RA). METHODS: Urinary porphyrin and precursor excretion, erythrocyte protoporphyrin, and antipyrine clearance, were measured in 6 patients with RA before and 10 weeks after commencement of intramuscular gold. RESULTS: Parameters of haem metabolism were unaffected by gold. While antipyrine clearance was not statistically changed after gold treatment, in 3 of the patients there was an average decrease in antipyrine clearance of 23%. CONCLUSION: Further studies examining RA patients at different time points are required to investigate further the possibility of reduced hepatic drug metabolising activity during prolonged treatment with gold. PMID- 10464559 TI - Expression of metalloproteinase-2 (gelatinase A) in labial salivary glands of patients with Sjogren's syndrome with HTLV-I infection. AB - OBJECTIVE: To determine whether gelatinase A (MMP-2) plays a significant role in the pathogenesis of Sjogren's syndrome (SS) with or without HTLV-I infection. METHODS: We examined 24 patients with SS (14 HTLV-I-seropositive and 8 HTLV-I seronegative). Labial salivary gland tissue samples were analysed immunohistochemically using anti-MMP-2 monoclonal antibodies. RESULTS: In normal salivary glands, MMP-2 expression was not detected. All biopsy samples of 8 SS patients with HTLV-I-associated myelopathy (HAM) and 3 of 6 HTLV-I-seropositive SS patients without manifestation of HAM stained positively for MMP-2. However, the other samples were negative for MMP-2. CONCLUSION: Our study showed the MMP-2 expression in labial salivary glands of HTLV-I seropositive SS patients, especially in all SS patients with HAM. The presence of MMP-2 in the salivary glands of these patients suggests that it may play a role in cellular infiltration and destruction in salivary glands of SS. PMID- 10464560 TI - Headache in systemic lupus erythematosus and its relation to other disease manifestations. AB - OBJECTIVE: To investigate headache in systemic lupus erythematosus (SLE) and its relation to other disease manifestations. METHODS: Clinical and laboratory variables of 148 SLE patients were prospectively recorded in a computed data base. RESULTS: The patients were divided into two groups. Group A consisted of patients who reported moderate to severe headache on at least two consecutive encounters, and Group B consisted of the remainder of the patients, with mild or no headache. The two groups did not significantly differ in age or in sex distribution. Patients in Group A suffered from more severe joint pain and inflammation, muscle pain, photosensitivity, mouth ulcers, fever and fatigue. They also had higher disease activity scores, and a higher number showed central nervous involvement. There were no significant differences between the two groups in any of the laboratory variables examined, nor in the proportion of patients with renal involvement. The prevalence of non-thromboembolic central nervous system (CNS) manifestations was 7.2%. The sensitivity of headache for the diagnosis of non-thromboembolic CNS manifestations was 90.9%, and the specificity was 29.2%. On logistic regression analysis, the total arthritis score, muscle pain, fatigue and photosensitivity were each found to be significantly independently related to headache. CONCLUSIONS: Headache is common in SLE, and in the majority of patients is related to musculoskeletal and constitutional disease manifestations. PMID- 10464561 TI - Bacterial infection presenting as cutaneous vasculitis in adults. AB - OBJECTIVE: To examine the frequency and clinical features of patients with bacterial infection presenting with biopsy-proven leukocytoclastic cutaneous vasculitis (CV) in a well-defined area of southern Europe (northwestern Spain). METHODS: A retrospective study of an unselected population of adult patients (age > 20 years) with biopsy-proven leukocytoclastic CV diagnosed at the Hospital Xeral-Calde (Lugo, Spain) was carried out from January 1988 through December 1997. Cutaneous vasculitis related to bacterial infection was considered if the vasculitis was confirmed by a skin biopsy showing leukocytoclastic vasculitis, if no drug intake was registered prior to the development of CV, and if bacteriologic evidence of infection was obtained. RESULTS: Four of 138 patients (2.9%) presenting with biopsy-proved CV were diagnosed with leukocytoclastic CV related to bacterial infection. Three patients (2 with bacterial endocarditis and 1 with meningococcemia) met the ACR criteria for the classification of hypersensitivity vasculitis. Another patient with bacterial endocarditis met the criteria for mixed cryoglobulinemia. All of them presented with palpable purpura, high or low grade fever, an elevated erythrocyte sedimentation rate and leukocytosis. CONCLUSION: Cutaneous vasculitis may be the presenting manifestation of bacterial infection. In this respect, rheumatologists should be aware of possible infectious causes of vasculitis, even though they are not common. PMID- 10464563 TI - Acute sacroiliitis as a manifestation of calcium pyrophosphate dihydrate crystal deposition disease. AB - While radiographic lesions of the sacroiliac joint (SIJ) are common in patients with calcium pyrophosphate dihydrate (CPPD) crystal deposition disease, they are rarely accompanied by clinical symptoms. We report the case of a 69-year-old woman who presented with an acute sacoiliitis and a linear calcification in the right SIJ on CT scan. The patient recovered well after intra-articular steroid injections. PMID- 10464562 TI - Rheumatoid resorptive arthropathy. PMID- 10464564 TI - Cogan's syndrome: unsuccessful outcome with early combination therapy. AB - Interstitial keratitis and vestibuloauditory symptoms (vertigo and hearing loss) are the typical signs of Cogan's syndrome, a rare inflammatory vascular disease. Signs of vasculitis in many organ systems may appear, among which neurologic problems are sometime predominant. The efficacy of glucocorticoids on the ocular and systemic symptoms is established, but their effect on hearing loss is unknown. We describe a case of Cogan's syndrome with neurological involvement in which early treatment with combination therapy (prednisolone and cyclosporin) failed to bring ear inflammation under control. PMID- 10464565 TI - The urokinase-type plasminogen activator system and inflammatory joint diseases. AB - Much evidence indicates that the urokinase plasminogen activator (u-PA), the urokinase receptor (u-PAR) and the serpin inhibitors are critical in cell invasion processes. The balance between u-PAR-bound u-PA and inhibitors modulate a pericellular proteolytic activity able to give "stop and go" signals to invading cells. The plasminogen activation system operates both directly and in concert with the matrix-metalloproteinase system. Direct interactions of u-PAR with vitronectin and integrins further regulate cell invasion. Another line of evidence suggests that u-PA-u-PAR interaction elicits chemotaxis, chemoinvasion and cell multiplication, events that do not require plasmin generation and therefore are referred to as "plasminogen-independent". Following the description of the main molecular and functional characteristics of the cell-surface associated plasminogen activation system, we discuss here the observations indicating a role of this system in many aspects of the rheumatic diseases, ranging from the infiltration of inflammatory cells into the affected joint, infiltration of synovial cells into the underlying cartilage, and remodeling of the cartilage itself. Evidence of the intraarticular cytokine- and growth factor dependent regulation of the components of the plasminogen activation system are presented in terms of the paracrine and autocrine regulation of receptor associated fibrinolysis. The roles of plasminogen-dependent and plasminogen independent u-PAR-associated events in various phases of joint inflammation are also discussed. A knowledge of these processes is required for the therapeutic utilization of antagonists of the u-PA/u-PAR system able to control the activity of proliferating and invading cells in inflammatory joint diseases. PMID- 10464566 TI - On beyond methotrexate treatment of severe juvenile rheumatoid arthritis. PMID- 10464567 TI - Spinal cord involvement in pediatric systemic lupus erythematosus: case report and literature review. AB - Transverse myelitis is a rare but serious complication of systemic lupus erythematosus (SLE). We describe the youngest patient with transverse myelitis ever recorded in SLE. Clinical improvement was observed after prednisone and cyclophosphamide therapy. The English language literature from 1966 to the present on the subject are reviewed. PMID- 10464568 TI - Recurrent monoarthritis in an 11-year-old boy with occult coeliac disease. Successful and stable remission after gluten-free diet. AB - A case of occult coeliac disease (CD) presenting with recurrent monoarthritis in a boy aged 11 years is reported. The case is unique due to the association of occult untreated CD and arthritis in childhood. Peripheral or axial arthritis as a first manifestation of occult CD has been described in adult patients, with an interval between the arthritis and CD of up to 15 years. In our case the interval between the appearance of arthritis and the diagnosis of CD was 2 years. The boy was asymptomatic for bowel disease and his nutritional status was normal. The diagnosis of CD was established using anti-gliadin (AGA) and anti-endomysium (EMA) antibody tests and was confirmed by small bowel biopsy. The introduction of a gluten-free diet resulted in the persistent remission of arthritis. As the treatment of CD-associated arthritis is based on dietary therapy, physicians should be alert to the possibility of occult CD in any child with arthritis of unclear origin. PMID- 10464569 TI - Hip bursitis in active polymyalgia rheumatica: report of a case. PMID- 10464570 TI - IL-6 and some natural inhibitors of chronic human inflammation in RA and SLE. PMID- 10464571 TI - Destructive arthritis of the shoulder triggered by trauma in a patient with polymyositis. PMID- 10464572 TI - Genetic testing and Alzheimer disease: recommendations of the Stanford Program in Genomics, Ethics, and Society. AB - Several genes associated with Alzheimer disease (AD) have been localized and cloned; two genetic tests are already commercially available, and new tests are being developed. Genetic testing for AD--either for disease prediction or for diagnosis--raises critical ethical concerns. The multidisciplinary Alzheimer Disease Working Group of the Stanford Program in Genomics, Ethics, and Society (PGES) presents comprehensive recommendations on genetic testing for AD. The Group concludes that under current conditions, genetic testing for AD prediction or diagnosis is only rarely appropriate. Criteria for judging the readiness of a test for introduction into routine clinical practice typically rely heavily on evaluation of technical efficacy. PGES recommends a broader and more comprehensive approach, considering: 1) the unique social and historical meanings of AD; 2) the availability of procedures to promote good surrogate decision making for incompetent patients and to safeguard confidentiality; 3) access to sophisticated genetic counselors able to communicate complex risk information and effectively convey the social costs and psychological burdens of testing, such as unintentional disclosure of predictive genetic information to family members; 4) protection from inappropriate advertising and marketing of genetic tests; and 5) recognition of the need for public education about the meaning and usefulness of predictive and diagnostic tests for AD. In this special issue of Genetic Testing, the PGES recommendations are published along with comprehensive background papers authored by Working Group members. PMID- 10464573 TI - Defining dementia: social and historical background of Alzheimer disease. AB - Though Alzheimer disease (AD) has been recognized as a distinct entity since 1907, scientific understanding of, and public interest in, the disease remained very limited until the 1970s. The perception of AD as a significant problem has been substantially affected by cultural and demographic changes and by interest group and federal government initiatives. The recognition of AD has transformed senility from an expected stage of life into a "disease." It has also increased fear both of the individual effects of having AD and of the social consequences of AD in the population. Both the biotechnology industry and AD activist organizations will play a role in the social implications of genetic testing for AD. PMID- 10464574 TI - Understanding genetic testing for Alzheimer disease: medical and epidemiological background. AB - To assess the potential benefits and risks of genetic testing for Alzheimer disease (AD), patients and families, health care providers, and policymakers must understand the medical and epidemiologic aspects of the disease. This paper provides a brief overview of the symptoms, progression, and etiology of AD, as well as of the prevalence and incidence of the disease. The established and the controversial risk factors thought to be associated with the development of AD are described. Current methods for diagnosing and treating AD are also reviewed. PMID- 10464575 TI - The personal and social consequences of Alzheimer disease. AB - Alzheimer disease (AD) has a drastic effect on the lives of those affected by the disease. Surprisingly little research has studied the personal and social consequences for AD patients. Many appear to go through six stages of change as the disease progresses, but any individual's path through the disease may vary substantially. AD also has a drastic effect on family members who serve as caregivers for people with the disease. These effects are much more studied and better documented. The burdens and, in some cases, benefits of the caregiver role differ from person to person. Race and ethnicity are among the variables that seem to play a role in the caregiver's response. The economic costs of AD are considerable, both for "formal" services, paid for in the money economy, and "informal" services, provided without monetary payment. The direct costs, both formal and informal, are largely, but not entirely, borne by the patient and the patient's family. The overall costs to society clearly amount to scores of billions of dollars. PMID- 10464576 TI - The genetics of Alzheimer disease and the application of molecular tests. AB - Two general classes of genes are associated with the development of Alzheimer disease (AD). The first group consists of genes that appear to cause AD when mutated, and the second category is composed of genes that are statistically associated with AD, depending on the inheritance of specific alleles. This paper reviews the current state of knowledge about the genetics of AD, and we then discuss the two molecular tests that are currently commercially available. These include a genetic test for mutations in the presenilin 1 (PS1) gene that can diagnose or predict a subset of early onset familial AD with a high degree of certainty. The value of the genetic test for the apolipoprotein (APOE) allele status is far less clear. Inheritance of the epsilon 4 allele is associated with an increased risk of AD at a population level, but APOE genotyping is inappropriate for prediction of future disease in an individual and offers only a marginal increase in diagnostic certainty when symptomatic individuals are tested. In the future, genetic tests may become more broadly applicable to the diagnosis and prediction of AD. However, the utility of such tests is currently limited to a small subset of individuals because in the vast majority of AD cases no clear genetic or environmental cause has been defined. PMID- 10464577 TI - Evaluation of genetic tests: APOE genotyping for the diagnosis of Alzheimer disease. AB - Many studies have now confirmed the association between inheritance of the epsilon 4 allele of the apolipoprotein E (APOE) gene and Alzheimer disease (AD). However, although the medical community holds the near-unanimous opinion that APOE genotyping should not be used for prediction in asymptomatic individuals, controversy remains about whether it should be used for diagnosis in patients who show signs of dementia. We assessed critically the recent clinical studies, on the basis of four criteria recommended to ensure safety and effectiveness of genetic tests. We also developed a formal framework for evaluating the usefulness of APOE genotyping using decision-theoretic principles. We conclude that neither the presence nor absence of an epsilon 4 allele provides diagnostic certainty, and the proper interpretation of either result in heterogeneous populations requires further investigation. The appropriate role of APOE genotyping among elements of a traditional assessment for AD has not been determined. Whether APOE genotyping provides sufficient information to change patient management decisions has not been determined. APOE genotyping presents foreseeable, significant psychosocial consequences for family members that must be weighed against any psychosocial benefits. Therefore, the diagnostic use of APOE genotyping outside research settings is premature until such testing is shown to be of practical value. PMID- 10464578 TI - Understanding probabilistic risk in predisposition genetic testing for Alzheimer disease. AB - The utility of genetic testing in preventing the onset of conditions such as Alzheimer disease (AD) depends upon categorizing individuals based on their "risk" of illness. Although no strategies to prevent the occurrence of AD have been proven effective, the promise of the new molecular medicine is based on the assumption that those "at risk" can be identified, counseled about their likelihood of developing a disease, and prescribed specific preventive interventions. We suggest that this paradigm of disease prevention through risk stratification has limitations that have not been fully explored. Within the fields of cognitive and health psychology, research has addressed how individuals understand the numeric presentation of uncertain future events; this research often focuses on the "perceptual pathology" of lay people. The uncertainty inherent in the risk estimates themselves is rarely considered, nor has research addressed how those risk estimates are created. There has been limited work to date focused on genetic risk assessment, and little is known about how individual might understand the meaning or consequences of being at genetic risk for AD. An emerging social science critique of concepts of risk in genetic medicine offers a broader perspective, examining the process of producing scientific risk estimates as well as the response of individual patients, such as the experience of embodied risk that is central to genetic testing. Making fully informed choices about genetic testing for AD may prove more difficult than our current model of ethical practice suggests. PMID- 10464579 TI - The application of medical decision analysis to genetic testing: an introduction. AB - The availability of genetic tests to diagnose or predict Alzheimer disease (AD) causes a shift in the way people think about the condition and how they assess the options available to them. Decision analysis in a quantitative approach for dealing with the uncertainties inherent in many medical decisions, including decisions about genetic testing. Decision analysis does not guarantee a good outcome, but aims to yield better overall average results by providing a framework for people to evaluate their options and minimize cognitive biases. We provide an overview of the decision analysis process, including the terms and tools commonly associated with it. We also use a recent example to demonstrate one way decision analysis has been applied to genetics in the medical literature. This paper is an introduction to subsequent papers that explore the specific question of whether decision analysis is a helpful tool for understanding the uncertainty inherent in probabilistic information about genetic risk for AD. PMID- 10464580 TI - Decision analysis and Alzheimer disease: three case studies. AB - Decision analysis may be useful to people facing Alzheimer disease (AD) decisions. The use of decision analysis in three such cases is reported. The first case involved a middle-aged person worried about early-onset AD and deciding whether to seek genetic testing. The analysis let the participant reject testing and consider innovative care options. The second case involved a middle aged person concerned about later-onset AD. The analysis for her was more complex, and led to the assignment of some limited value on genetic testing for her. The third case revolved around a caregiver's treatment decisions for a patient with severe AD. It led her to recognize the importance of factors she had not previously considered. In each of the three cases, the intensive process of decision analysis appears to have improved the subject's decision. PMID- 10464581 TI - A philosophical critique of decision analysis as a tool in genetic testing. AB - Decision analysis has been proposed as a method to improve the quality of decisions made by individuals facing choices about genetic testing for Alzheimer disease (AD) and other conditions where information about risk is highly uncertain. This paper provides a philosophical critique of two schools of decision analysis, explaining the conceptual limitations inherent in each approach. A central difference between the two approaches is their stance toward the ontological status of patient preferences and values. Are an individual's preferences and values about genetic testing simply assessed or extracted during decision analysis or are they created during the decision analysis itself? How closely can preferences conform to the axioms of rational choice theory? A case study of one individual's experience with decision analysis demonstrates the strengths and weaknesses of decision analysis in the context of genetic testing for AD. PMID- 10464582 TI - Surrogate decision making for genetic testing for Alzheimer disease. AB - Genetic tests are commercially available for the purpose of aiding in the differential diagnosis of Alzheimer disease among patients with dementia. Such patients often lack the mental capacity to consent to or reject such testing. If genetic testing is to be undertaken, it is important legally and ethically to consider who should participate in the decision to test. State law and the patient's previously expressed wishes will determine which individual should serve as the surrogate decision maker. Other family members should be included in the discussion of the decision, and their assent to the surrogate's decision should be sought. PMID- 10464583 TI - Personal identity, advance directives, and genetic testing for Alzheimer disease. AB - The authors consider current theories of personhood and personal identity as they relate to the Alzheimer disease (AD) patient, and evaluate their usefulness in clinical contexts. Differing conclusions about the weight advance directives should be given in decision making for seriously demented patients suggest that the philosophical presuppositions that underlie various positions can have implications for the care of such patients. PMID- 10464584 TI - The law, adolescents, and the APOE epsilon 4 genotype: a view from Canada. AB - The presence of an apolipoprotein E (APOE) epsilon 4 allele may be a risk factor for the severity of the consequences of head trauma, both short-term and long term. If this proves to be true, genetic testing for APOE status might be encouraged, or required, to minimize the costs of participation in high-contact sports, such as ice hockey, soccer, rugby, and boxing, where head injury is likely to occur. Under some circumstances, testing minors for this purpose might be justifiable. This would raise complex ethical and legal questions about respective powers and rights of adolescents and parents. The wishes of the "nearly competent" adolescent not to be tested should override parental desires for testing, although such a conclusion has only limited warrant in Canadian law. If a mature adolescent wishes testing, but the parents do not, testing may be permissible--although Canadian law is again unclear. The physician should not be able to inform the parents of the test results without the adolescent's consent, nor should the adolescent be forced to disclose those results. PMID- 10464585 TI - Special issues in genetic testing for Alzheimer disease. AB - Genetic testing for Alzheimer disease (AD) raises two issues that are, thus far, unusual. First, genetic testing of dementia patients for diagnostic purposes necessarily leads to information of some predictive significance for the patient's family members. What information about the test results should be disclosed to whom needs careful consideration. Testing for a patient's apolipoprotein E (APOE) allele status was used, both in research and clinically, as a predictor of cardiovascular risk long before it was known to be associated with AD risk. Whether and how information about the newly understood AD implications of the test should be provided to those who were tested for cardiovascular risk needs attention, as does the general problem of new, and possibly less benign, meanings for old genetic test results. PMID- 10464586 TI - The psychological impact of genetic testing for Alzheimer disease. AB - Information regarding one's genetic risk for a particular disease might effectively inform medical, financial, and reproductive decisions and perhaps promote established risk reduction behaviors. However, genetic testing may also lead to significant levels of anxiety, depression, or other forms of distress, especially when psychological coping strategies and social reserves are not adequate to manage positive or inconclusive results. This paper focuses on the psychological impact of predisposition genetic for Alzheimer disease (AD). We present stress and coping models that capture the essence of this impact and discuss the potential role of counseling and follow-up interventions. The discussion draws primarily from the experiences of professionals working with other diseases, and seeks to expand that experience into the AD arena. In addition, we emphasize two issues we believe need additional attention in the psychological literature regarding genetic testing and counseling: (i) the psychosocial vulnerability of individuals being tested and, (ii) the sociocultural context of the AD patient and their family members. PMID- 10464587 TI - Genetic testing, Alzheimer disease, and long-term care insurance. AB - Patients with Alzheimer Disease (AD) often need long-term care, which is poorly covered by private or governmental health insurance. Private long-term care (LTC) insurance is increasingly being marketed to provide some financial support for long term care. LTC insurance does cover AD, but the insurers will not sell policies to people who have already been diagnosed with AD. As a result, LTC insurance might be particularly attractive to individuals who are at heightened risk for developing AD. This makes the issues surrounding predictive genetic testing for AD and the use of test results in determining insurance premiums and eligibility of great concern to patients, clinicians, insurers, ethicists, and patient advocate groups. The paper examines the complexities of these issues. PMID- 10464588 TI - Public understanding of genetics and Alzheimer disease. AB - It is generally thought that public understanding of science is inadequate. The definitions of "the public," "understanding," and "science" all need careful consideration in this formulation, with a greater focus on who should understand what and for what purposes. Improved public understanding depends on experts being reflective about the limits of their own expertise and the different needs of the many potential consumers of genetic knowledge. Media coverage of genetic discoveries has been extensive in recent years, but this coverage is often not useful and may even be misleading to families at risk for genetic disease. The Internet provides an alternative source of information as well as an interactive forum, one that has been widely used by persons interested in Alzheimer disease (AD). Internet-based activities hold great promise for providing interested members of the public with useful information and facilitating more substantive dialogue among physicians, researches, and laypersons regarding the genetic aspects of AD. PMID- 10464589 TI - High school genetics education and Alzheimer disease. AB - Improved and updated human genetics education, including Alzheimer disease (AD) awareness and education, is urgently needed. National, state, and local standards for science education agree that human genetics, biotechnology applications, and the social and ethical issues raised by modern technology need to be taught in high school science using hands-on and inquiry methods of teaching. High school science courses are the last opportunity for most individuals to learn human genetics. There are an increasing number of new and successful human genetics curriculum materials and inservice teacher education programs at the secondary school level aligned with national and state science education standards. These curricula and teacher education programs can be enhanced by collaborative partnerships of geneticists, genetics professionals, biotechnology scientists and technical personnel, and science educators, several of which are in successful operation. Because human genetics involves families and generations, genetics education tied to AD may provide a unique opportunity to educate two generations, both students and parents, to the many medical, personal, family, community, and cultural issues of human genetics and genetic conditions. Implementing human genetics and AD education provides a recipe for accurate, relevant, sustainable and exciting teaching and learning for all involved. PMID- 10464590 TI - Genetic risk assessment and insurance. PMID- 10464591 TI - The use of genetic tests and genetic information by life insurance companies: does this differ from the use of routine medical information? AB - Questions regarding insurance companies' access to and use of genetic test results and genetic information have been raised since the advent of the Human Genome Project. The ability to place applicants of similar risks in groups, a process known as underwriting, is critical to the availability and affordability of individually underwritten life, disability income, and long-term care insurance. This paper focuses on how life insurance companies use medical information and particularly genetic test results and other genetic information in the risk assessment process. Issues and concerns raised about differences between routine medical information and genetic information are addressed and policies with regard to privacy and confidentiality are presented. PMID- 10464592 TI - Genetic testing and health insurance practices: an industry perspective. PMID- 10464593 TI - Genetic discrimination: who is really at risk? PMID- 10464594 TI - Genetic information must remain private to prevent discrimination, spur research. PMID- 10464595 TI - Developing genetic privacy legislation: the South Carolina experience. AB - The availability of presymptomatic and predisposition genetic testing has spawned the need for legislation prohibiting health insurance discrimination on the basis of genetic information. The federal effort, the Health Insurance Portability and Accountability Act (HIPAA) of 1996, falls short by protecting only those who access insurance through group plans. A committee of University of South Carolina professionals convened in 1996 to develop legislation in support of genetic privacy for the state of South Carolina. The legislation prevents health insurance companies from denying coverage or setting insurance rates on the basis of genetic information. It also protects the privacy of genetic information and prohibits performance of genetic tests without specific informed consent. In preparing the bill, genetic privacy laws from other states were reviewed, and a modified version of the Virginia law adopted. The South Carolina Committee for the Protection of Genetic Privacy version went a step further by including enforcement language and excluding Virginia's sunset clause. The definition of genetic information encompassed genetic test results, and importantly, includes family history of genetic disease. Our experience in navigating through the state legislature and working through opposition from the health insurance lobby is detailed herein. PMID- 10464596 TI - Accurate, high-throughput "snapshot" detection of hMLH1 mutations by two dimensional DNA electrophoresis. AB - The human genome and related projects have resulted in the isolation of a rapidly growing number of genes that cause susceptibility to human cancer. With rare exception, these genes are large and have disease-associated mutations scattered along the length of the genes. Therefore, the development of accurate and cost efficient mutation detection tests that can scan entire genes singly or in combination is warranted. hMLH1, encoding a mismatch repair enzyme, is a susceptibility gene for hereditary nonpolyposis colon cancer syndrome. This gene comprises 19 exons; mutations are scattered, typical of many susceptibility genes. Here, we present a strategy that combines extensive PCR multiplexing and two-dimensional DNA electrophoresis (Two-Dimensional Gene Scanning, TDGS) to scan accurately for mutations that lie within the exons and splice junctions of hMLH1. All target fragments, designed to have optimal melting characteristics, were prepared in a two-stage PCR--a four-plex long-distance PCR followed by short PCR in two multiplex groups of 10 and 11 amplicons. The mixture of amplicons was subjected to two-dimensional electrophoresis: separation by size in the first dimension and by melting characteristics in the second. Using this design, 41 samples containing known hMLH1 sequence variants or no alterations were blindly subjected to TDGS. All mutations were detected; there were no genuine false positive or false-negative results. These results confirm that TDGS is a generally applicable, rapid, accurate, and reproducible mutation detection technology that would serve large-scale molecular epidemiologic studies as well as clinical molecular diagnostic purposes. PMID- 10464597 TI - Neonatal screening for Duchenne muscular dystrophy: a novel semiquantitative application of the bioluminescence test for creatine kinase in a pilot national program in Cyprus. AB - The objectives of this study were to evaluate a novel semiquantitative application of the bioluminescence test for screening newborns for Duchenne muscular dystrophy (DMD) and to use this technique in a pilot national program. The study was performed on the island of Cyprus, which provides ideal conditions for maximizing the prevention rate due to the small size of the country, the well defined population, and the high degree of awareness of the public concerning genetic diseases. Guthrie spots were obtained through the national screening center for phenylketonuria and congenital hypothyroidism. The bioluminescence method for measuring creatine kinase (CK) in dried blood spots was adapted for use in a semiquantitative way. During the first 6 years of the program (1992 1997), we screened 30,014 samples and found 43 with initially high CK values. We were able to obtain repeat specimens in 35 cases. Of the repeat samples, 30 were found to have normal activity, giving a false-positive rate of 0.10%. Five boys had persistent CK elevations and were confirmed to be DMD or Becker (BMD) cases by DNA analysis and/or dystrophin analysis. The semiquantitative application of the bioluminescence assay of CK that we have introduced has proved to be a fast and reliable method for screening large numbers of samples for DMD. It has a low rate of false positives, which compares favorably with that of other DMD screening programs. Although it is early to evaluate its impact fully, the program seems to be bringing about the anticipated benefits to affected families. PMID- 10464598 TI - The role of physician preferences in the choice of amniocentesis or chorionic villus sampling for prenatal genetic testing. AB - Our objective was to determine the effect of physician preferences, as well as physician demographic, obstetric, and practice-related factors, on the choice of prenatal test made by their patients. We studied preferences for prenatal outcomes for 372 pregnant women who either chose amniocentesis (AMN) (n = 288) or chorionic villus sampling (CVS) (n = 84) for the indication of maternal age. We also studied preferences for these outcomes for the 92 physicians that referred them for testing. Preferences were assessed using written scenarios and were measured on linear rating scales. According to patients, the choice of prenatal test was made entirely or mostly by the physician in 14% of cases and was shared equally between patient and physician in 37% of cases. After adjustment for patient preferences, physician concern about spontaneous abortion of a normal fetus after CVS (odds ratio 0.71; CI, 0.48-1.05; p = 0.08), and a limb reduction (LRD) birth after CVS (odds ratio 0.85; CI, 0.68-1.05; p = 0.12), tended to decrease their patients' odds of choosing CVS, but the results were not statistically significant. No other physician preference, and no physician demographic, obstetric, or practice-related factor, influenced patient test choice. We conclude that after taking patient preferences into account, physician preferences and practice-related factors did not emerge as significant determinants of the choice of prenatal test made by their patients. It remains possible, however, that physician concern about spontaneous abortion and about LRD increase the likelihood of their patients choosing AMN over CVS. PMID- 10464600 TI - Communicating chromosome rearrangements and their outcomes using simple computer generated color ideograms. AB - Simple computer-generated color ideograms (CGCIs) conforming to ISCN (1995) are presented, not only to represent partial karyotypes, but also to depict reproductive outcomes displaying potential imbalances in a form that is easy to understand. This approach has been found to facilitate communication greatly between cytogeneticist, clinician, and patient, and could be easily adopted by most centers. PMID- 10464599 TI - A simple system for automated two-dimensional electrophoresis: applications to genetic testing. AB - Two-dimensional gene scanning (TDGS) is a method for mutation detection based on two-dimensional electrophoretic separation of PCR-amplified DNA fragments according to size and base pair sequence. To facilitate two-dimensional electrophoresis without manual interference, a simple three-chamber unit was designed on top of an existing vertical electrophoresis instrument. Application of this system to genetic testing is demonstrated by the performance of a complete two-dimensional mutational scanning assay of one of the hereditary nonpolyposis colon cancer syndrome susceptibility genes, hMLH1. PMID- 10464601 TI - Deleterious mutations of both BRCA1 and BRCA2 in three siblings. PMID- 10464603 TI - Is the hemochromatosis gene a modifier locus for cystic fibrosis? AB - The variable clinical manifestations of cystic fibrosis (CF) suggest the influence of modifier genes. For example, meconium ileus is present in approximately 10-15% of neonates with cystic fibrosis; however, the genetic and, or environmental factors that determine whether an individual will develop this complication have not been determined. We propose the HFE gene as a candidate modifier locus for CF based on (1) the suggestion of an association between the HLA loci and CF phenotypes; (2) the location of the HFE gene near the HLA loci and; (3) the similarity between the gastrointestinal manifestations of hereditary hemochromatosis and CF. We have determined the frequency of the C282Y and H63D mutations in a group of 89 CF patients who were homozygous for delta F508 and for whom meconium ileus status was known. The carrier frequency of C282Y among the CF patients with meconium ileus was significantly different from that of our unaffected control group (19.4% versus 7.7%). However, the difference between the meconium ileus and the nonmeconium ileus groups was not significant (19.4% versus 10.3%). There was no difference in the frequency of the H63D among the three groups that were studied. These data are suggestive of a relationship between the development of meconium ileus or other gastrointestinal diseases in CF and the HFE gene. Further study of a larger group of patients is warranted. PMID- 10464602 TI - Genetic polymorphism of CYP2E1, ADH2, and ALDH2 in Mexican-Americans. AB - The major enzymes involved in the metabolism of ethanol are alcohol dehydrogenases (ADH) and aldehyde dehydrogenase (ALDH). Some of the isozymes of ADH are expressed polymorphically. Studies investigating a causal link between ADH expression and alcoholic liver disease (ALD) have so far produced conflicting results. The cytochrome P450 2E1 (CYP2E1) represents a second enzyme that can metabolize ethanol. Although normally a minor route of metabolism, its role in chronic alcoholics may be proportionately greater than in nonalcoholics because CYP2E1 is inducible by ethanol. An Rsa I restriction fragment length polymorphism (RFLP) in the 5'-flanking region of the CYP2E1 gene has been identified. Studies have shown that the mutant allele demonstrates greater transcriptional rate, protein level, and enzyme activity when compared with the wild-type allele. The association between the Rsa I site polymorphism and ALD has been reported. In this report, we examined the genotypes of ADH2(2), ALDH2(2), and CYP2E1 in a group of healthy subjects of Mexican-American descent. The ADH2(2) and ALDH2(2) frequencies are 6% and 0%, respectively, which are similar to those which have been reported for Caucasians. In contrast, the Rsa I allele frequency of the CYP2E1 gene is 16%, which is significantly higher than in Caucasians. The high RsaI allele frequency found in Mexican-Americans suggests that it might play a role in the development of ALD in this rapidly growing minority population where ALD is common. PMID- 10464604 TI - Disease genes and chromosomes: disease maps of the human genome. Chromosome 22. PMID- 10464605 TI - Tay-Sachs disease carrier screening: a model for prevention of genetic disease. AB - Tay-Sachs disease (TSD) is an autosomal-recessive, progressive, and ultimately fatal neurodegenerative disorder. Within the last 30 years, the discovery of the enzymatic basis of the disease, namely deficiency of the enzyme hexosaminidase A, made possible both enzymatic diagnosis of TSD and heterozygote identification. In the last decade, the cloning of the HEXA gene and the identification of more than 80 associated TSD-causing mutations has permitted molecular diagnosis in many instances. TSD was the first genetic condition for which community-based screening for carrier detection was implemented. As such, the TSD experience can be viewed as a prototypic effort for public education, carrier testing, and reproductive counseling for avoiding fatal childhood disease. More importantly, the outcome of TSD screening over the last 28 years offers convincing evidence that such an effort can dramatically reduce incidence of the disease. PMID- 10464606 TI - High frequency of a common Bloom syndrome Ashkenazi mutation among Jews of Polish origin. AB - Bloom syndrome (BS) is an autosomal recessive disorder characterized by small stature, immunodeficiency, chromosomal instability, and a predisposition to different types of cancer. Although extremely rare in the general population, BS is seen in about 1 in 48,000 Ashkenazi Jews. Mutation analysis of seven Ashkenazi BS probands has shown that all were homozygous for the same mutation in the BLM gene: 2281delATCTGAinsTAGATTC, also known as blmAsh. This finding, along with the increased incidence of BS among Ashkenazi Jews, suggests a founder effect for BS in this population. The purpose of this study was to determine the frequency of blmAsh mutation carriers in a randomly sampled Ashkenazi Jewish population in Israel. The initial study group included 1,613 Ashkenazi Jews who were referred for routine DNA screening tests (cystic fibrosis, Gaucher, Canavan, fragile X). None had a family history of BS. A group of 552 non-Ashkenazi Jews served as controls. Mutation analysis was performed by PCR amplification followed by analysis of a specific BstN1 restriction site, created by the blmAsh mutation. All positive carriers were confirmed by direct sequencing. Sixteen blmAsh carriers were detected among 1,613 Ashkenazi Jews (1 in 101), compared to none among 552 non-Ashkenazi individuals. In this study, Ashkenazi Jews of biparental Polish descent had a significantly higher proportion of the blmAsh mutation (1 in 37) compared to Ashkenazi Jews of non-Polish descent. These results provide further evidence that a founder effect is responsible for the increased incidence of Bloom syndrome among Ashkenazi Jews, particularly those of Polish descent. PMID- 10464607 TI - Asymptomatic Gaucher disease implications for large-scale screening. AB - Gaucher disease, the most prevalent genetic disorder among Ashkenazi Jews, is characterized by significant phenotypic heterogeneity. Because seven mutations account for more than 96% of the disease alleles in this population, large-scale screening is feasible, and genotyping is commercially available. To date, only 400 Gaucher patients have been diagnosed in Israel, although 2,500 patients are predicted by gene frequency; hence, there is the probability that those uncovered by population screening will prove to be very mild or asymptomatic since these patients have generally escaped medical attention until now. We studied objective and subjective aspects in 68 asymptomatic/very mild patients, followed for a mean of 2.6 years, to assess the implications of identifying many more such patients if large-scale screening were to be implemented. We found there were no medically significant changes in disease severity during follow-up. The patients' subjective reports of their general health status were similar to those of normal subjects, and there were no significant changes in quality of life during follow up. Our findings suggest that asymptomatic/mild Gaucher patients do not require frequent monitoring; there is no justification for "prophylactic" enzyme therapy in this group, and, hence, no rationale for large-scale screening in the Ashkenazi population. Furthermore, one needs to be concerned with the adverse effects of labeling large numbers of asymptomatic individuals as affected because of the potential for stigmatization and discrimination. PMID- 10464608 TI - Establishing criteria for a carrier detection panel: lessons from the Ashkenazi Jewish model. PMID- 10464609 TI - Genetic testing for breast-ovarian cancer susceptibility: a regional trial. AB - To evaluate receptivity to testing for a genetic susceptibility to breast-ovarian cancer, information is needed on the response when the offer is open to all qualifying women in a given region. To qualify in this trial, a woman who had not had breast or ovarian cancer had to have at least two first-degree relatives or one first- and one second-degree relative with breast and/or ovarian cancer, whereas a woman who had had breast or ovarian cancer had to have at least one first-degree relative with breast or ovarian cancer and a first- or second-degree relative without cancer willing to be tested. Of 140 women qualifying and interested enough to return questionnaires requesting baseline information, 111 were referred by their physician and 29 were identified from a regional tumor registry. Of these 140, 112 came for pretest education and 98 of these chose to be tested. Thus, the acceptance rate was 70% for all those returning baseline questionnaires, but 88% for those interested enough to come for pretest education. The most common reasons for accepting testing were to take extra precautions if a mutation were found (42.9%) and to determine if offspring were at risk (24.5%). The most common reasons for declining were anxiety and absence of specific interventions. Factors predicting who chose testing were years of education (p < 0.005) and family closeness (p < 0.02). Fourteen deleterious BRCA1 or BRCA2 mutations were found in 13 of the 87 families actually tested. If the criteria for testing had been three or more affected family members rather than two or more, the number of families tested would have been reduced by 46%, but the number of families found to have a deleterious mutation would have been reduced by only 9%. PMID- 10464610 TI - An efficient protocol for rare mutation genotyping in a large population. AB - We introduce a method to efficiently detect rare mutations for individual subjects in a large population by pooling samples and retesting subgroups of positive pooled samples. We conducted computer simulations of this method and discovered that it seems efficient for mutation prevalences less than 0.1, regardless of the number of samples. The simulations also indicate that splitting the pooled samples into three to five subgroups at each level is optimal. The expected number of necessary tests and relative efficiency of this method are given, by mutation prevalence and sample size. PMID- 10464611 TI - Discovery of children's carrier status for recessive genetic disease: some ethical issues. AB - Knowledge of one's carrier status for recessive genetic diseases is useful primarily in making marital and reproductive decisions. These decisions are peculiarly the private domain of the young adults who are dating, mating, and forming new families. The privacy of these decisions may be compromised when parents know the carrier status of their children. Thus, the practice of sharing that information with the parents of fetuses, babies, and minor children ought to be discouraged, out of respect for the autonomy and privacy of these children when they become adults. PMID- 10464612 TI - Genetic protocols review by Institutional Review Boards at National Cancer Institute-designated cancer centers. AB - Absent a clear and enforceable national policy on cancer genetic testing and genetic research, the responsibility may currently be on institutional review boards to regulate those activities at their institutions through protocol approval or disapproval. The survey reported here was carried out to gain information on National Cancer Institute-designated cancer center policies governing institutional review board review of genetic protocols and the preparedness of institutional review boards to review genetic protocols. Thirty five responses (63% response rate) were received, of which 30 were evaluable. Twenty-four responders reported that they believed that there is a need for research that may lead to improved review of genetic research and genetic testing protocols. Only 14 responders felt adequately informed of current and developing issues and legislation relative to genetic testing and research. Seven responders reported that their cancer centers require an institutional review board-approved protocol for genetic testing activities. Five responders reported that their cancer centers have a formal written policy that guides institutional review board review of genetic testing protocols. About half of the responders reported that their cancer centers have no formal written policy that guides institutional review board review of genetic research protocols. Only three responders reported that institutional review board members receive formal training to prepare them to evaluate all of the issues associated with genetic protocols. We conclude that greater effort needs to be made to establish uniform policy governing cancer genetic testing and genetic research and greater effort should be made to prepare institutional review boards formally for the review of genetic-related protocols. PMID- 10464613 TI - A multiplex ARMS test for 10 cystic fibrosis (CF) mutations: evaluation in a prenatal CF screening program. AB - In Maine, prenatal screening for cystic fibrosis (CF) is offered through primary care providers. Cheekbrush (buccal) samples are routinely tested for eight mutations by multiplex PCR amplification of five exons, followed by dot-blot hybridization with pooled allele-specific oligonucleotides (ASO). The ASO methodology is widely used and effective, but somewhat time and labor intensive when applied to CF carrier testing or couple-based prenatal screening in the general pregnant population. Amplification Refractory Mutation System (ARMS) is an improvement of the PCR that allows rapid detection of mutations involving single base changes or small deletions/insertions. In this study, two multiplex ARMS reactions are used to test for 10 common CF mutations. Clinical evaluation of the ARMS test includes a retrospective study of 140 banked samples (54 cell line, proficiency testing, and buccal controls; 86 clinical buccal samples) with known CF genotype (57 with CF mutations, 83 no mutation), followed by a prospective trial in which 309 buccal samples are analyzed con-currently using both methods. The success rate of the ARMS test in buccal lysates is comparable to the ASO method; all CF mutations are successfully identified. For testing nonsterile buccal lysates with low DNA concentrations, optimized performance in the ARMS method is obtained using Amplitaq Gold polymerase. The ARMS method developed is easy, rapid (1 day), and avoids the need for ASO probe labeling, dot blotting and autoradiography. This study provides further evidence that ARMS methodology is suitable for clinical CF mutation analysis. PMID- 10464614 TI - Simultaneous detection of five mutations in the steroid 21-hydroxylase gene using nested allele-specific amplification. AB - Congenital adrenal hyperplasia due to deficiency of steroid 21-hydroxylase (CYP21) is most frequently due to mutations that arise from the nearby CYP21 pseudogene. The mechanism involves either unequal crossing over, which deletes part of the CYP21 functional gene, or gene conversion which puts a mutation from the pseudogene into the functional gene. We have devised an assay to rapidly screen for five known mutations that are due to gene conversion within an 1,800 bp region of the CYP21 gene--I172N, V281L, Q318X, R356W, and a cluster of mutations in exon 6 (I236N, V237E, M239K). This method is based on a set of nested allele-specific polymerase chain reactions done simultaneously in one tube, for which we suggest the acronym NASA, for nested allele-specific amplification. The assay is capable of detecting the mutations individually as well as all combinations of mutations tested. PMID- 10464615 TI - A simple VNTR-PCR method for detecting maternal cell contamination in prenatal diagnosis. AB - The effectiveness of variable number tandem repeats (VNTRs) was evaluated in the detection of maternal cell contamination. Nonradioactive PCRs were performed on 30 sets of prenatal tissue using VNTRs as primers. The combination of two VNTRs (YNZ22 and APOB) provided information on all 30 cases, distinguishing maternal fetal genotype patterns and detecting maternal cell contamination in 5 of 30 prenatal cases. The amplification of these two VNTRs does not require radioactive or fluorescence labeling, and a small gel electrophoresis is sufficient to see the maternal-fetal genotype pattern. By this method, detection of maternal cell contamination in prenatal tissues can be obtained in 1 day, without the use of expensive instruments, thus providing DNA laboratories a very sensitive, rapid, and simple proof pretest on all prenatal tissues before performing the final genetic diagnostic testing. PMID- 10464617 TI - Disease genes and chromosomes: disease maps of the human genome. Chromosome 17. PMID- 10464616 TI - Allele drop-out can occur in alleles differing by a single nucleotide and is not alleviated by preamplification or minor template increments. AB - The ability to analyze the genetic material of single cells by the PCR opens up new prospects for diagnostics. Because only two copies of the genetic template are available for amplification, a problem that frequently arises when examining heterozygous loci in single cells is allele drop-out (ADO). ADO results from the preferential amplification of one of a pair of heterozygous alleles, in which the other allele is totally under-represented. In examining single cells from carriers heterozygous for beta-thalassemia mutations, we have found ADO to occur in alleles differing by a single nucleotide, where either the normal or the mutant genotype was absent. We have found that ADO is not overcome by either increasing the amount of DNA template to 20 pg or by primer extension preamplification (PEP), but rather that the best diagnostic accuracy is obtained by examining multiple single cells and basing a diagnosis on the combined results of such an examination. PMID- 10464618 TI - Genetic diseases and testing in Ashkenazi Jews: Part 1. PMID- 10464619 TI - Gaucher disease: gene frequencies and genotype/phenotype correlations. AB - Gaucher disease is the most prevalent lysosomal storage disease and has its highest incidence in the Ashkenazi Jewish population. Over 100 mutant alleles have been identified in affected patients, but four alleles, termed N370S, L444P, 84GG, and IVS2, have significant frequencies in this population. In affected patients, genotype data show that the presence of a single N370S allele is diagnostic of the type 1 or nonneuronopathic variant, whereas the L444P/L444P genotype is highly associated with neuronopathic variants in the Caucasian population. Large screening studies also indicate a significant underestimation (approximately two-fold) of the prevalence of the N370S/N370S genotype in the affected Ashkenazi Jewish patient population. These results indicate that the N370S/N370S genotype provides a necessary but not sufficient condition for the development of the Gaucher disease phenotype. The genotype/phenotype correlations and gene frequencies have significant impact on genetic counseling of at-risk couples and the future need for therapy of affected patients. PMID- 10464620 TI - Niemann-Pick disease: mutation update, genotype/phenotype correlations, and prospects for genetic testing. AB - Niemann-Pick Disease (NPD) is an autosomal recessive lysosomal storage disorder caused by a deficiency of acid sphingomyelinase (ASM). NPD occurs in two forms, neuronopathic Type A and nonneuronopathic Type B. The incidence of Type A NPD is highest among Ashkenazi Jews. Type B NPD is more common in non-Jews but has been reported in Ashkenazi Jews. Different mutations in ASM are presumed to be responsible for the different NPD phenotypes. Three mutations are predicted to account for > 95% of all Type A NPD chromosomes among Ashkenazi Jews (L302P, R496L, fsP330). Based on limited screens for these mutations among Ashkenazi Jews, a carrier frequency for Type A NPD of 1:90 is reported for this population. Less is known about mutations responsible for Type B NPD, although one mutation (delta R608) has been identified in both Ashkenazi Jews and non-Jews. Screening of the Ashkenazi Jewish population to detect > 95% of NPD carriers can be accomplished with a four-mutation panel that includes L302P, R496L, fsP330, and delta R608, the three predominant Type A mutations and one recurrent Type B mutation. PMID- 10464621 TI - Canavan disease: diagnosis and molecular analysis. AB - Canavan disease, spongy degeneration of the brain, is an autosomal recessive disorder with increased prevalence among Ashkenazi Jews. The biochemical marker for this disease is increased levels of N-acetylaspartic acid, due to the defective enzyme, aspartoacylase. This discovery allowed for accurate diagnosis of the disease. The gene for aspartoacylase has been cloned and two mutations have been found to be responsible for Canavan disease among Ashkenazi Jewish patients in 98% of the cases. Molecular analysis of healthy Jewish individuals for these mutations has resulted in an unexpectedly high carrier frequency for Canavan disease among Jews. Therefore, carrier testing of the Jewish population is possible and indicated. PMID- 10464622 TI - Fanconi anemia: genetic testing in Ashkenazi Jews. AB - Fanconi anemia (FA) is an autosomal recessive disorder characterized clinically by progressive pancytopenia, diverse congenital abnormalities, and a predisposition to malignancy, particularly acute myelogenous leukemia (AML). Hypersensitivity of FA cells to the clastogenic effect of crosslinking agents such as diepoxybutane (DEB) is used as a diagnostic criterion, because phenotypic heterogeneity makes clinical diagnosis difficult. Studies of genetic heterogeneity have shown that there are at least five different complementation groups, FA-A through FA-E. Overall, FA-A is the most prevalent group, accounting for 60%-65% of all FA. The FAA gene, which maps to chromosome 16q24.3, was recently isolated and methods for molecular diagnosis of FA-A are currently being developed. The first FA gene to be isolated (FAC) maps to chromosome 9q22.3; FA-C accounts for 10%-15% of FA. A variety of mutations and polymorphisms have been described in FAC. The most common of these is IVS4 +4 A-->T, which is the only FAC mutation found in Ashkenazi Jewish FA patients and their families. This mutation has not been found in any affected individual of non-Jewish ancestry. The carrier frequency of the IVS4 mutation was found to be 1 in 89 (1.1%; 95% confidence interval 0.79% to 1.56%) in an Ashkenazi Jewish population, whereas no carriers were identified in an Iraqi Jewish population, which represents the original gene pool of the Jews. We have developed amplification refractory mutation system (ARMS) assays for FAC mutations, which provide a means of rapid, nonradioactive genetic testing. These assays have been used to assign FA patients to Group C, to provide rapid carrier testing and prenatal diagnosis for FA-C families. PMID- 10464623 TI - Cystic fibrosis in Jews: frequency and mutation distribution. AB - The incidence of cystic fibrosis and the frequency of disease causing mutations varies among different ethnic groups and geographical regions around the world. The Jewish population is comprised of two major ethnic groups. Ashkenazi and Non Ashkenazi. The latter is further classified according to country of origin. An extreme variability in the disease frequency (from 1:2400-1:39,000) was found among the different Jewish ethnic groups. In the entire Jewish CF population, only 12 mutations were identified that altogether enable the identification of 91% of the CF chromosomes. However, in each Jewish ethnic group, the disease is caused by a different repertoire of a small number of mutations. In several ethnic groups, there is a major CFTR mutation that accounts for at least 48% of the CF chromosomes. High proportion of the CF chromosomes can be identified in Ashkenazi Jews (95%), Jews originating from Tunisia (100%), Libya (91%), Turkey (90%), and Georgia (88%). High frequencies of CFTR mutations were found among infertile males with CBAVD who might not have additional CF clinical characteristics. Of the Jewish males with CBAVD, 77% carried at least one CFTR mutation. The 5T mutation is the major mutation in Jewish CBAVD affecteds accounting for 32% of the chromosomes among Ashkenazi Jews and 36% among the non Ashkenazi Jews. Five additional CFTR mutations, W1282X (12%), delta F508 (9%), N1303K (3%), D1152H, (5%)), and R117H (1%) were identified among Ashkenazi Jews with CBAVD. Only two mutations, delta F508 and R117H, were found among non Ashkenazi males with CBAVD. An increased frequency of the 5T allele was also found among Jewish patients with atypical CF presentation, 18% in Ashkenazi, and 10% in non-Ashkenazi Jews. In summary, we present the required information for genetic counseling of Jewish families with typical and atypical CF and for carrier screening of healthy Jewish individuals. PMID- 10464624 TI - BRCA1 and BRCA2 mutations in Ashkenazi Jewish families with breast and ovarian cancer. AB - The strongest risk factors currently known for inherited predisposition to breast and ovarian cancer are mutations in BRCA1 and BRCA2. Two mutations in BRCA1 and one mutation in BRCA2 have been identified that are present to a particularly high degree in the Ashkenazi Jewish population due to ancient founder effects. To clarify the role of ancient and novel BRCA1 and BRCA2 mutations in the Ashkenazi Jewish population, families with a strong history of breast and ovarian cancer were examined. Seventeen Ashkenazi Jewish families with four or more breast or ovarian cancers were analyzed for ancient and novel mutations in BRCA1 and BRCA2. Ancient mutations existed in 9 families; 7 had the BRCA1 185 del AG mutation, 1 had BRCA1 5382 ins C, and 1 had BRCA2 6174 del T. A novel mutation, BRCA2 6425 del TT, was discovered in 1 of the remaining 8 families. Seven families with four or more cases of breast and ovarian cancer cannot be accounted for by either the ancient or novel mutations. Therefore, ancient mutations in BRCA1 and BRCA2 are present in approximately half of Ashkenazi Jewish families in this series, suggesting the possibility of novel mutations, either in BRCA1, BRCA2, or in currently unidentified gene(s), responsible for the remainder. PMID- 10464625 TI - Prevalence of recurring BRCA mutations among Ashkenazi Jewish women with breast cancer. AB - The BRCA1 mutations 185delAG and 5382insC and the BRCA2 mutation 6174delT have been detected in a significant proportion of Ashkenazi Jewish women with early onset breast cancer. A group of 236 Jewish women with breast cancer was screened for the presence of these alterations. Mutations were detected in 25.0% (59/236). Among women with breast cancer diagnosed at or before the age of 45, the prevalence of these mutations was 29.1% (42/144). Among women diagnosed with breast cancer after age 45, mutations were noted in 18.5% (17/92). Among women with a family history of breast or ovarian cancer, the likelihood of detecting a mutation was 32.1% (53/165). BRCA1 185delAG was the most common mutation overall (40/236, 16.9%). The ratio of BRCA1 185delAG to BRCA2 6174delT was 4.0 in women with early-onset breast cancer and 1.3 in women with breast cancer diagnosed after age 45. Clinical features such as age at diagnosis, family history of breast or ovarian cancer, bilateral breast cancer, and personal history of breast and ovarian cancer increase the likelihood of detecting mutations among Ashkenazi women with breast cancer. The yield of testing is low in the absence of any of these features. PMID- 10464626 TI - Cystic fibrosis carrier population screening: a review. AB - Population screening for cystic fibrosis (CF) carriers, now possible because of the cloning of the CFTR gene, merits evaluation because CF is common, serious, and without satisfactory treatment, and because prenatal diagnosis is available. Clinical trials of CF population carrier screening are reviewed. These trials have involved pregnant women, adults of both sexes of reproductive age, or adolescents. Schools, the usual setting for screening programs for adolescents, provide an excellent opportunity for a formal educational component and for comprehensive coverage of the population, but compared to a health-care setting, may entail subtle coercion and may compromise confidentiality. In the case of adults, many say they prefer screening before conception but do not see a physician for evaluation before conception and providers find screening more readily accomplished in the setting of a prenatal visit. Two large U.S. studies of prenatal screening with quite different subject populations and health-care settings encountered few of the adverse outcomes originally predicted for CF carrier population screening. PMID- 10464628 TI - Disease genes and chromosomes: disease maps of the human genome. Chromosome 18. PMID- 10464627 TI - Introduction to PCR/OLA/SCS, a multiplex DNA test, and its application to cystic fibrosis. AB - The field of medical, molecular diagnostics has grown rapidly over the last few years, becoming increasingly informative to both clinician and patient. As genes associated with specific diseases have been discovered and sequenced, many genotype-phenotype relationships have been defined. For those genetic diseases with associated, defined, gene mutations, sophisticated DNA diagnostic tests are being developed. As an example, the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene, is associated with Cystic Fibrosis (CF). We have developed a new molecular diagnostic technology, PCR/OLA/SCS, and applied it first to the diagnosis of CF. Test design in the field of molecular diagnostics must consider such characteristics as specificity, sensitivity, ease and speed of protocol, multiplex capacity, and cost. PCR/OLA/SCS addresses these requirements. Polymerase Chain Reaction (PCR) is widely used in both diagnostics and research. We have combined well established PCR technology with Oligonucleotide Ligation Assay (OLA) and Sequence-Coded Separation (SCS), two relatively new technologies. PMID- 10464629 TI - Breast cancer susceptibility testing: Part 1. PMID- 10464630 TI - Mutation testing of early-onset breast cancer genes BRCA1 and BRCA2. AB - BRCA1 and BRCA2, genes predisposing to early-onset breast cancer, have been isolated and are characterized for mutation spectrum, risks of cancer, and function. The different methodologies to screen for mutations in BRCA1 and BRCA2 are briefly discussed including DNA-based methodologies and potential new assays. The numbers and types of mutations identified to date are described, including the problems of ascribing risk to missense mutations. Recurring, possibly founding mutations have been identified in several populations including Ashkenazi Jews, Icelanders, Swedes, and African Americans. From population-based studies, estimates are that 6%-10% of breast cancers are due to mutations in BRCA1 and BRCA2. Knowledge of mutation status raises additional questions including the interpretation of negative tests and the risks of breast and other cancers associated with positive test results. PMID- 10464631 TI - Genetic testing for breast cancer susceptibility: frequency of BRCA1 and BRCA2 mutations. AB - Genetic testing for breast cancer susceptibility became a reality after two cancer predisposition genes, BRCA1 and BRCA2, were identified. Mutations in these two genes were predicted to account for 85% to 90% of hereditary breast and ovarian cancer syndromes. We present results of mutation analysis of the coding sequence of these two genes in 110 consecutive non-Jewish breast cancer patients with a positive family history of breast and/or ovarian cancer. The individuals were identified in various cancer risk evaluation centers in the country. Twenty two (20%) mutations in the BRCA1 gene and 8 mutations (7%) in the BRCA2 gene were detected. We also analyzed 52 Ashkenazi Jewish breast cancer patients for mutations in the BRCA1 and BRCA2 genes. Eleven Jewish individuals (21%) carried either one of the two common mutations, 185delAG and 5382InsC, in the BRCA1 gene and 4 individuals (8%) had the 6174delT mutation in the BRCA2 gene. The frequency of mutations in BRCA genes in affected people in this ethnic group was not significantly different from the non-Jewish population. On further analysis, the data demonstrate that neither age of onset nor phenotype of the disease had any significant predictive value for the frequency of mutations in these genes. These data confirm the lower prevalence of mutations in either of the BRCA genes in clinical families when compared to high-risk families used for obtaining linkage data in a research setting. PMID- 10464632 TI - Genetic counseling for BRCA1/BRCA2 testing. AB - Genetic counseling plays a key role in the BRCA1/BRCA2 testing process. The initial genetic counseling encounter will determine the appropriateness of the test by collecting a detailed family history and determining the likelihood that the family has a BRCA1 or BRCA2 mutation. Once the test is offered, then genetic counseling discussions center around the possible test results, implications of the results to the patient and other relatives, and risks and benefits of testing. The goal of this pre-test genetic counseling session is to ensure that patients have sufficient information with which to make a decision about being tested. At results disclosure, individuals can learn their results along with information about cancer risks and medical management options. Follow-up genetic counseling services can provide continued support and help arrange consultations with other medical care providers as needed. All clinical BRCA1/BRCA2 testing programs should include pre- and post-test genetic counseling. PMID- 10464633 TI - The 22q11.2 deletion: screening, diagnostic workup, and outcome of results; report on 181 patients. AB - A submicroscopic deletion of chromosome 22q11.2 has been identified in the majority of patients with the DiGeorge syndrome, velocardiofacial syndrome, conotruncal anomaly face syndrome, and in some patients with isolated conotruncal cardiac anomalies, Opitz G/BBB syndrome, and Cayler cardiofacial syndrome. We have evaluated 181 patients with this deletion. We describe our cohort of patients, how they presented, and what has been learned by having the same subspecialists evaluate all of the children. The results help define the extremely variable phenotype associated with this submicroscopic deletion and will assist clinicians in formulating a management plan based on these findings. PMID- 10464634 TI - PCR assay for screening patients at risk for 22q11.2 deletion. AB - Deletions of 22q11.2 have been detected in the majority of patients with DiGeorge, velocardiofacial, and conotruncal anomaly face syndromes by either cytogenetic analysis, fluorescence in situ hybridization (FISH), or Southern blot hybridization. However, these techniques may not be the most efficient or cost effective means of screening large numbers of "at-risk" patients. Therefore, we developed a PCR assay to assess a patient's likelihood of having a 22q11.2 deletion based on homozygosity at consecutive markers in the DiGeorge chromosomal region. The sensitivity and specificity of PCR screening were evaluated in a cohort of cardiac patients. We conclude that a PCR-based assay is a reliable and efficient means of identifying which patients are at greatest risk for a 22q11.2 deletion and should have FISH studies to confirm their deletion status. PMID- 10464636 TI - Genetic testing for male infertility: a postulated role for mutations in sperm nuclear matrix attachment regions. AB - Numerous reports have suggested that disturbances in nuclear condensation may result in male infertility. This notion has been supported by the observation of infertile individuals with a decrease or absence of the male sperm-specific chromatin packaging protamine proteins. To date, no correlation between the absence of protamine proteins and a mutation within the coding regions of the protamine genes has been documented. To address this issue, PCR-based mutation scanning analysis has been performed across the human male haploid expressed PRM1 ->PRM2-->TNP2 domain in several oligozoospermic infertile individuals. This analysis identified a candidate mutation in a region of contact with the sperm nuclear matrix from 2 of 5 affected individuals. This is the first report of a mutation scan covering the entire PRM1-->PRM2-->TNP2 locus in affected individuals. PMID- 10464635 TI - Dystrophin point mutation screening using a multiplexed protein truncation test. AB - We report here the first use of a multiplexed protein truncation test for the high throughput screening of dystrophin point mutations. We have developed a substantially more robust and efficient procedure incorporating large savings in cost which uses muscle biopsy or lymphocyte total RNA as the template. The entire dystrophin open reading frame is screened in only five overlapping fragments using a long RT-PCR strategy to amplify dystrophin cDNA in excess of 3.7 kb. These five fragments are uniquely transcribed and translated in vitro in a single multiplexed reaction containing magnesium ions to reduce nonspecific internal initiation of translation. We have used this system to analyze mutations in 11 Duchenne muscular dystrophy patients (10 unrelated) with previously uncharacterized mutations. A single truncating mutation was identified in all patients, which was confirmed at the genomic level. Multiplex PTT provides the most efficient method for point mutation screening in this large gene and has potential applications to several disease genes with a significant proportion of truncating mutations. PMID- 10464637 TI - Who should provide genetic education prior to gene testing? Computers and other methods for improving patient understanding. AB - As numerous new gene tests are introduced into clinical practice, patients have a growing need for accurate and comprehensive information about the risks and benefits of gene testing. However, in the changing healthcare environment, it is not clear who will provide such information because genetic counselors are scarce and their services are not widely utilized, and primary care providers lack time and expertise in genetics. Interactive computers may help fill the information gap. We review a variety of educational modalities for providing patient education and argue that interactive computers have potential advantages over other educational methods for providing information and promoting informed consent to genetic testing. Finally, some questions for further research are raised. PMID- 10464638 TI - The Oncormed approach to genetic testing. AB - This report describes a commercial laboratory's novel approach to providing genetic testing services to detect BRCA1 mutations in persons with hereditary breast/ovarian cancer. The approach involves the use of institutional review board (IRB)-approved protocols as a paradigm for conducting genetic testing in a commercial setting. We discuss the rationale for this approach and the key elements of the protocol. In addition, we provide data on the first 6 months of implementation of the protocol in 32 clinical sites. A phased testing approach was used, consisting of an allele-specific oligonucleotide assay for the 8 most common BRCA1 mutations, a protein truncation test of exon 11, and direct sequencing of the remaining regions of the gene. Data are presented on the yield of mutation carriers by category of family history and by stage of analysis. PMID- 10464639 TI - Disease genes and chromosomes: disease maps of the human genome. Chromosome 19. PMID- 10464640 TI - Methylation analysis of the fragile X syndrome by PCR. AB - The fragile X syndrome is predominantly caused by a large expansion of a CGG trinucleotide repeat in the promoter region of the FMR1 gene, which is associated with methylation and downregulation of transcription. The molecular diagnosis of this disorder is based on repeat size and methylation analysis of the FMR1 gene usually by Southern blot analysis. We describe a PCR-based method for the analysis of methylation of the FMR1 gene, which involves bisulfite treatment of DNA prior to amplification. Fifty-two normal and 48 affected, premutation, or mosaic males were analyzed in a blinded study by this method. A prospective study of 30 males suspected of fragile X was also performed. Amplification specific for the methylated FMR1 sequence was readily observed in all individuals with a full mutation, whereas all normal and premutation individuals showed only amplification-specific for the unmethylated sequence, thus, allowing affected and unaffected males to be distinguished. A full mutation in the presence of mosaicism was also detectable by this method. Methylation-specific PCR appears to be a rapid and reliable tool for the diagnosis of fragile X males. PMID- 10464641 TI - Evaluation of genotype data in clinical risk assessment: methods and application to BRCA1, BRCA2, and N-acetyl transferase-2 genotypes in breast cancer. AB - Associations of numerous susceptibility genes with disease risk have been reported. However, objective methods have not been developed to evaluate the conditions under which translation of knowledge about susceptibility genotypes may be clinically informative. We describe and apply a statistical approach to evaluate when genotype information may be clinically informative in disease risk assessment. We estimate an interval of cumulative cancer incidences where it may be appropriate to use these genes in disease risk assessment. We also estimate the magnitude of a log odds ratio (H) that measures genotype-disease association. We illustrate this method with three breast cancer susceptibility genotypes: population screening data evaluating the 185delAG mutation at BRCA1 and the 6174delT mutation at BRCA2 in a Ashkenazi Jewish population, and case control data for the slow acetylation genotype at the N-acetyl transferase 2 (NAT2) gene in combination with smoking. Knowledge of the 185delAG mutation in BRCA1 (HdelAG = 3.42; 95% CI: 3.04, 3.79) or the 6174delT mutation in BRCA2 (HdelT = 1.98; 95% CI: 1.16, 2.30) can be clinically informative in distinguishing individuals who are and are not at breast cancer risk in populations with cumulative breast cancer incidences of > or = 4% and > or = 13%, respectively. NAT2 genotypes alone are much less clinically informative in predicting breast cancer risk (HNAT2 = 0.10). However, knowledge of both heavy smoking 20 years ago and NAT2 genotype is a more clinically informative predictor of postmenopausal breast cancer risk with HNAT2 = 2.19, when the cumulative breast cancer incidence in the target population is at least 31%. These results indicate that knowledge of the 185delG mutation-status may be clinically informative even in populations with low cumulative breast cancer incidences, whereas the 6174delT mutation and NAT2 genotypes may only be clinically informative in a population with higher cumulative breast cancer incidence. The proposed approach can be used to objectively evaluate the conditions under which susceptibility genotypes may be applied for risk assessment or genetic screening. PMID- 10464642 TI - An allelic variant at the ATM locus is implicated in breast cancer susceptibility. AB - We have tested a simple procedure, disease association by locus stratification, for identifying breast cancer patients with pathogenetic allelic variants at several candidate loci. The strategy was based on the assumption of epistatic interactions of the candidates. We analyzed 66 independent cases from sib pairs affected with breast cancer that had previously been collected during an investigation of pathogenetic-allele-sharing at the HRAS1 mini-satellite locus. An exon 24 polymorphism of ATM, substituting arginine for proline was associated with breast cancer in these cases with an overall odds ratio of 4.5 (95% confidence interval, 1.2-20.5, nominal p = 0.02, 2-tail Fisher exact test). In the presence of a rare HRAS1 allele, the odds ratio increased to 6.9 (95% CI, 1.2 38.3, p = 0.03). Thus, our procedure identified at least one allelic variant of ATM associated with breast cancer, and indicated that the ATM locus may interact with HRAS1. PMID- 10464643 TI - Automated detection of prevalent mutations in BRCA1 and BRCA2 genes, using a fluorogenic PCR allelic discrimination assay. AB - Mutations in the genes BRCA1 and BRCA2 account for 5%-10% of familial early onset breast cancer. Identification of these mutations allows molecular diagnosis for breast cancer susceptibility. A high through-put automated PCR allelic discrimination assay (ADA) was developed to detect the prevalent mutations in these genes. Two allele specific oligonucleotides (ASO) are directly used in the PCR reaction, in both of which the fluorescent reporter and quencher dyes are attached to the 5' and 3' ends, respectively. During PCR, fluorescence is generated after cleavage of the annealed ASO by the 5' nuclease activity of Taq polymerase. The wild-type BRCA sequence is distinguished from the mutant sequence by the differential fluorescence emission of two different reporter dyes. The sensitivity of ADA is at the level of a single cell following a nested PCR. Eighty-six patient samples can be analyzed for each mutation in 15-min post-PCR without the need for radioactivity, gel electrophoresis, or membrane blotting/hybridization. PMID- 10464644 TI - Mutation analysis and genetic service: the construction and use of national confidential databases of mutations and pedigrees. AB - The development of rapid mutation screening procedures allows the detection of mutations in large populations. This is particularly useful for inherited diseases of high mutational heterogeneity, such as haemophilia A and B, because the analysis of the very many different natural mutants clearly defines the features that are important to the function of the relevant gene and gene product. Furthermore, the characterization of the mutation in an index person from each affected family may lead to the construction of confidential databases of mutations and pedigrees that allow optimization of genetic service. We report how, motivated by the aforementioned concepts, we have planned and introduced in the UK a national strategy to optimize genetic service in both haemophilias and, in particular, we describe the principles that have guided us. PMID- 10464645 TI - Caught in the middle again: professional ethical considerations in genetic testing for health risks. AB - Genotyping tests for molecular mutations associated with clinical syndromes increasingly allow clinicians to identify health risks before clinical problems occur, sometimes making prevention possible. The clinical use of these tests, however, can create moral problems for families and serious health policy challenges for communities. Those issues, in turn, complicate the professional ethics of genetic testing and counseling. Investigators working with families in gene-identification studies have already encountered these complications in the research setting and their experiences may help in the development of a clinical ethic for genetic testing. In addition to questions about the predictive significance of particular genotypes, three other sets of ethical considerations are becoming important to professional decision-making about genetic testing: the psycho-social impact of testing, the patient's privileges with respect to testing, and the potential for effective prevention following testing. Underlying all these considerations are basic lessons from the research setting on how clinicians should interpret genetic testing for patients, given the heavily deterministic meanings that they have been taught thus far. PMID- 10464646 TI - Detection of single-nucleotide polymorphisms with the WAVE DNA fragment analysis system. AB - As the Human Genome Project is generating an avalanche of genetic information, molecular researchers and clinical practitioners are setting new criteria for evaluation of the links between newly discovered gene mutations and human disorders. These requirements necessitate the development of highly accurate and yet rapid automated systems for genetic testing. We describe the detection of a single nucleotide polymorphism (SNP) on the human Y chromosome with the fully automated, sensitive, and rapid system WAVE designed for DNA fragment analysis. This new technology, based on temperature-modulated liquid chromatography and a high-resolution matrix, offers new dimensions to the molecular biology research. The versatility of the WAVE makes the equipment a universal molecular biology stations for clinical and research facilities. The key aspects to setting the operating parameters are discussed. PMID- 10464647 TI - Implications of molecular diagnostic testing in families with hereditary pancreatitis. AB - Hereditary Pancreatitis (HP), is an autosomal dominant trait, which presents with recurrent attacks of abdominal pain, and is the most common cause of chronic relapsing pancreatitis in children. In addition to recurring episodes of intense epigastric pain, patients have nausea, vomiting, and anorexia, and typically show elevated serum amylase levels during the acute episode that can rapidly decline in convalescence. Complications of long-standing disease include features of chronic pancreatitis, such as pancreatic pseudo-cyst, exocrine and endocrine failure, parenchymal calcification, and pancreatic cancer. A large family from Virginia, which was originally studied by Katwinkle and Lapey in 1973, was re ascertained through a new proband. Linkage studies in this family mapped the gene to the 7q35 region, with similar results being reported simultaneously by two other groups. A pathogenic G to A transition mutation in exon 3 of the cationic trypsinogen (CT) gene, which had previously been mapped to this region, was found both in our family as well as other families from North America. Many other conditions can produce abdominal symptoms that are often mis-attributed to the disease in HP families. An affected member of our family in whom the mutation was confirmed by direct sequencing of exon 3 of the cationic trypsinogen gene requested diagnostic testing on his 4-year-old son because of onset of severe abdominal pain and vomiting. Screening for the mutation in this child did not reveal the pathogenic G to A change. These results prevented unnecessary invasive diagnostic procedures and treatment in this child. The pre-symptomatic testing of high risk individuals could, thus, have a significant impact on the well being of both affected and normal family members. PMID- 10464648 TI - Technical pitfalls encountered in PCR quantification using microsatellites. AB - Quantitative PCR has proved useful for different purposes, including the detection of particular genetic changes, such as deletions and duplications in several inherited disorders. Using patients with the known duplication mutation for Charcot-Marie-Tooth disease Type 1A as examples, the importance of selecting informative microsatellite loci and proper electrophoretic conditions so as to eliminate potential sources of error in quantitative PCR studies is discussed. PMID- 10464650 TI - Disease genes and chromosomes: disease maps of the human genome. PMID- 10464649 TI - Characterization of the GALC gene in three Japanese patients with adult-onset Krabbe disease. AB - Krabbe disease, a neurodegenerative disorder of autosomal recessive inheritance, is caused by mutations in the galactosylceramidase (GALC) gene. However, its clinical manifestations in terms of time of onset and severity are heterogeneous. Thus, elucidation of the relationship of symptoms to the site and type of mutation is important, both for an understanding of the etiology of the disease and for diagnostic purposes. We examined the genomic structure of the GALC gene in three unrelated adult-onset Krabbe disease patients. One patient was homozygous for an Ile66Met mutation. Another patient who appeared to express only one mutated mRNA species was, in fact, a compound heterozygote for an Ile66Met mutation and a nonsense mutation, Tyr354ter. Because the allele with the nonsense mutation was not detectable by mRNA analysis, a rapid degradation of the mRNA caused by premature chain termination was suggested. The third patient who carried two inactiving mutations--Leu618Ser and a second resulting in exon 6 skipping--was also found to carry an intronic mutation, IVS6 + 5G > A. Transfection experiments using a GALC mini-gene proved that this intronic mutation was the cause for the exon 6 skipping. PMID- 10464651 TI - Prepregnancy genetic testing for age-related aneuploidies by polar body analysis. AB - Current practice for prevention of chromosomal aneuploidies involves prenatal screening and termination of pregnancy, a procedure that is not universally acceptable. We introduced prepregnancy genetic testing by sampling and fluorescence in situ hybridization (FISH) analysis of the first and second polar body (PB), to avoid fertilization and transfer of embryos resulting from aneuploid oocytes. In 395 in vitro fertilization (IVF) patients of advanced maternal age, the first and second PBs were removed following their extrusion from oocytes and studied by FISH, using probes specific for chromosomes 13, 18, and 21, to detect and avoid the transfer of oocytes with common aneuploidies. Overall, 3,651 oocytes obtained from 598 IVF cycles were available for FISH analysis, with 2,952 showing interpretable FISH results (80.9%). The analysis revealed 1,271 (43.1%) oocytes with aneuploidy, which were excluded from transfer and subjected to follow-up FISH analysis to confirm PB diagnosis in the cleavage or blastocyst stage embryos. Only embryos originating from 1,681 aneuploidy-free oocytes were transferred back to patients, resulting in 119 pregnancies overall, from which 78 healthy children have already been born, 35 were spontaneously aborted, and 16 are ongoing, after confirming PB diagnosis by prenatal diagnosis. The results demonstrate that PB-based preimplantation diagnosis may be used for prepregnancy screening in women with age-related risk for common aneuploidies. PMID- 10464652 TI - Denaturing HPLC-identified novel FBN1 mutations, polymorphisms, and sequence variants in Marfan syndrome and related connective tissue disorders. AB - Marfan syndrome (MFS), a common connective tissue disorder, is caused by fibrillin-1 (FBN1) mutations that are scattered throughout the gene and are largely unique to individual families. Mutation detection in this large gene of 65 exons is a considerable technical challenge. To develop an efficient method capable of identifying all possible mutations in this gene, we have explored the use of a novel denaturing high-performance liquid chromatography (DHPLC) system. This technique compares two or more chromosomes as a mixture of denatured and reannealed PCR amplicons. Under partially denaturing conditions, heteroduplexes can be separated from homoduplexes. A panel of 94 DNA samples from individuals with MFS or related connective tissue disorders was screened exon-by-exon by this method. A total of 66 unique heteroduplex profiles was identified. Sequencing of the amplicons detected 37 novel and two previously reported mutations, as well as 15 novel and 10 known polymorphisms or unique sequence variants that are probably of no clinical significance. Of the 34 mutations found in definitive MFS cases, 16 were identified in the 21 samples that had not been screened before (76% detection rate) and 17/40 (43%) were in samples previously screened by other mutation detection methods. In 32 individuals with MFS-related phenotypes, five FBN1 mutations were identified (16%). Our results demonstrate the power of the DHPLC method to detect FBN1 mutations. It should be applicable for mutation screening in any gene in a large population. PMID- 10464653 TI - Branchio-oto-renal syndrome: identification of novel mutations, molecular characterization, mutation distribution, and prospects for genetic testing. AB - The branchio-oto-renal syndrome (BOR) is an autosomal dominant disorder characterized by branchial clefts, preauricular sinuses, hearing loss, and renal anomalies. The BOR gene, EYA1, on chromosome 8q13 has recently been cloned and mutations have been identified. In this study, we have analyzed the sites of mutations in the EYA1 gene in BOR patients to determine the spectrum of mutations. We have identified two missense mutations and have compared all the mutations reported to date in the EYA1 gene. In total, 20 mutations have been described, the majority of which are clustered in the carboxy-terminal region of the gene. The clinical features of the BOR individuals have also been compared to determine if the nature of the mutation correlates with the type and severity of the clinical symptoms. Most of the mutations arose de novo and, other than the clustering in carboxy-terminal exons 9-16, no mutation hot spots have been identified. These results provide the basis for molecular genetic testing that will help in the clinical evaluation and genetic counseling of members of BOR families. PMID- 10464654 TI - Comparison of enzyme mismatch cleavage and chemical cleavage of mismatch on a defined set of heteroduplexes. AB - Two mutation detection methods based on the cleavage of mismatched heteroduplexes were compared and evaluated. These techniques, chemical cleavage of mismatch (CCM) and enzyme mismatch cleavage (EMC), have the advantages over other available methods of being able to detect and localize mutations in relatively large fragments of DNA (> or = 1 kb). We have constructed clones that enable us to create heteroduplexes of 500 bp, 1 kb, and 1.5 kb and have assessed each of the methods over a range of criteria. Both were able to detect and localize all four types of single-base mismatch and insertion/deletions of 1-5 bp. Whereas EMC was efficient at detection of insertion/deletions in a broad size range of fragments and has the advantage over CCM of using no hazardous chemicals, in our hands it has not been sufficiently robust that we felt confident to consider it for diagnostic use in its present form. CCM using hydroxylamine was efficient over the entire range of fragment sizes tested and using potassium permanganate with tetraethylammonium chloride was efficient up to 1 kb. PMID- 10464655 TI - Diagnosis of ataxia telangiectasia with the glycophorin A somatic mutation assay. AB - There are no widely applied definitive laboratory tests for the diagnosis of ataxia telangiectasia (AT). We, and others, have previously reported significantly elevated levels of in vivo somatic mutation in blood samples from known AT patients, observations that might form the basis for a useful prospective laboratory test for confirmation of a clinical diagnosis of AT. In the present case, a 4 1/2-year-old black female was suspected of having AT based on ataxic gait and chronic upper respiratory infections. Blood work-up showed low IgG2 and elevated alpha-fetoprotein (AFP), consistent with the AT phenotype. Her peripheral blood karyotype was normal, however, with no spontaneous breakage observed among 100 solid stained metaphases. Lymphocytes from AT patients often show elevated levels of chromosome rearrangement, especially at sites of immunoglobulin and T-cell receptor genes. Therefore, a blood sample was analyzed with the glycophorin A (GPA) in vivo somatic mutation assay. The GPA assay detects and quantifies the phenotypically variant erythrocytes resulting from loss of heterozygosity for the MN blood group. The patient had a 10-fold increased frequency of variant erythrocytes with a phenotype consistent with simple loss of the N allele, which is characteristic of AT. In addition, the variant cell distribution for this patient showed three other, more qualitative hallmarks of AT: a normal frequency of allele loss and duplication events, a unique ridge of cells of intermediate phenotype between the normal and mutant peaks, and evidence of similar ongoing mutational loss of the M allele. Together with clinical data, these distinctive qualitative and quantitative features of the GPA assay allow for a diagnosis of AT with a projected accuracy of 95%. Therefore, we suggest that the GPA assay, which can be performed on < 1 ml of blood and completed in less than a day, be considered as a confirmatory laboratory test for a clinical diagnosis of AT. PMID- 10464657 TI - Estimated contribution of known ataxia genes in ataxia patients undergoing DNA testing. AB - We estimated the relative contributions of known ataxia genes (SCA1, 2, 3, 6, 7 and X25) in the patient population sent to our DNA diagnostic laboratory for diagnostic testing. Approximately 28% of these patients had an abnormal triplet repeat expansion in one of these ataxia genes (3% for SCA1, 8% for SCA2, 11% for SCA3/MJD, 2% for SCA6, 3% for SCA7, and 1.5% for X25). The lack of abnormal repeat expansions in the majority of ataxis patients tested suggests that the molecular defects associated with most ataxia cases are currently undetermined and that this population includes both familial and sporadic cases. In contrast, of the patients submitted for genetic testing for Friedrich's ataxia (FRDA), 44% (69/157) showed at least one expansion in the X25 gene, indicating that FRDA accounts for a significant proportion of the recessively inherited ataxias and appears to have a high rate of accurate clinical diagnosis. On the basis of our DNA studies, we propose a comprehensive and efficient approach for molecular analysis of ataxia patients. PMID- 10464656 TI - Genetic screening of targeted subpopulations: the role of communal discourse in evaluating sociocultural implications. AB - Targeting socially identifiable subpopulations for genetic screening entails the risk of stigmatizing them. The potential for such harm should be considered before programs are initiated. There is an emerging consensus that targeted subpopulations should be actively involved in evaluating these risks. A process of communal discourse engages the community in discussions that reflect both public and private sociocultural contexts in which individual decisions about screening will be made. This allows the subpopulation to address the collective implications of testing in a culturally appropriate way. Communal discourse was used to evaluate the collective implications of genetic testing in two Native American communities. We found that private social units were more influential than public units in reaching communal consensus, that local sociocultural issues were of more concern than were general issues such as employment and insurance discrimination, and that heterogeneity within a subpopulation may be just as significant a consideration in designing a targeted screening program as diversity between subpopulations. Heterogeneity is constructed by using a dichotomy between community-specific and biomedical health representations and practices. How genetic screening is socially constructed using a community's existing dichotomy may be central to its success. PMID- 10464658 TI - Practice variations in biochemical screening for Down syndrome. AB - The American College of Obstetricians and Gynecologists has recommended that when using biochemical screening for Down syndrome, the cut-off selected for counseling the patient should be consistent with the Down syndrome risk at which the obstetrician routinely offers prenatal cytogenetic diagnosis on the basis of maternal age alone. They also state that multiple marker testing in women over the age of 35 years cannot be recommended for routine Down syndrome screening as an equivalent alternative to offering prenatal cytogenetic diagnosis. The American College of Medical Genetics has issued similar statements. We wanted to determine current practices in regard to these suggestions. A questionnaire was mailed to members of the Society of Perinatal Obstetricians asking: (1) At what maternal age do they offer counseling/amniocentesis? (2) Do they adhere to the Colleges' recommendations with regard to counseling/amniocentesis? (3) Do they change the risk for women age 35 years or over based on serum screening? Results showed that (1) Most members currently identify attaining the age of 35 years as a separate risk factor. (2) There were wide variations in the risk quoted for women 35 years old. (3) Many members used a different risk for their biochemical screening cut-off. (4) 74% of the members would increase the age-related risk for women over 35 years based on serum screening and 49% would decrease this risk based on serum screening. Among members, there were wide variations in the risks quoted; the biochemical screening cut-off used; and the use of serum screening in women over the age of 35 years. PMID- 10464659 TI - Arylsulfatase A pseudodeficiency: altered kinetic and heat-inactivation properties. AB - Arylsulfatase A (ASA) pseudodeficiency (PD) was described in clinically healthy individuals with ASA-deficient activity. To confirm that the PD individual in the present study is homozygous for the PD allele without any other mutations, direct solid-phase sequencing was done and the two A-to-G transitions--one at the third N-glycosylation site (N350S) and the other at the first polyadenylation signal (ATTAAC to AGTAAC)--were identified. No other mutations were detected in the entire coding region nor in the intron-exon boundary region of the ASA gene in the PD cells. Kinetic studies to compare the partially purified ASA from controls to that from a homozygote (PD allele) were carried out using p-nitrocatechol sulfate (p-NCS) as a substrate. The apparent Km for the control ASA was 0.6 mM and for the PD enzyme 2.0 mM (p < 0.01). The heat inactivation at 60 degrees C revealed 50% inactivation within 90 min for control ASA and 28 min for PD ASA. At 65 degrees C, the 50% inactivation was reached at 18 min for the control and at 8.5 min for the PD. These results document the decreased affinity of ASA toward p NCS and increased heat inactivation from a PD individual. Western blot analysis following SDS-PAGE and isoelectric focusing revealed differences in both the molecular weight and the isoelectric point between the control ASA and that of the PD allele. To the best of our knowledge, this is the first report showing the altered properties of ASA from a PD homozygote. PMID- 10464660 TI - Prenatal sexing and sex determination in infants with ambiguous genitalia by polymerase chain reaction. AB - Sex determination in both fetuses and infants with ambiguous external genitalia usually necessitates time-consuming and costly karyotyping. We propose a simple, rapid, and reliable method of prenatal and postnatal sex determination by means of the PCR, a technique currently used to identify gender for forensic purposes. DNA was extracted from 20 samples of whole blood from infants with ambiguous genitalia and from five samples of amniotic fluid. Three markers were amplified from each specimen: a Y chromosome alphoid repeat sequence and two homologous genes, amelogenin (AME) and zinc finger protein (ZFP). All three were detected in under 10 hr. A comparison of the results obtained with those of cytogenetic analysis of the same samples showed a perfect sex match, demonstrating that this PCR technique provides an accurate means of determining gender. PMID- 10464661 TI - Prenatal diagnosis in congenital contractural arachnodactyly. AB - Congenital contractural arachnodactyly (CCA) is a heritable connective tissue disorder caused by defects in the gene encoding fibrillin-2 (FBN2). People with CCA typically have a marfanoid habitus, flexion contractures, severe kyphoscoliosis, abnormal pinnae, and muscular hypoplasia. Because of the relative infrequency of the syndrome and its generally mild to moderate severity, prenatal diagnosis had not previously been sought. Here we report prenatal diagnosis in a family with CCA. Because the course of the disease in the proband was rather severe, she had requested genetic counseling as early as age 17. She delayed childbearing until prenatal diagnosis for CCA became possible. This decision was supported by her mother and later her husband. Because she shared the same genotype with her husband, genetic linkage analysis of this family did not alter the a priori 50% risk of having an affected child. The possibility of unambiguously ascertaining the affected status of a fetus homozygous for the tested FBN2 marker was sufficient for the family to pursue prenatal diagnosis. This case strongly points to the importance of informed decisions now that genetic testing is becoming commonplace. PMID- 10464662 TI - Fine mapping of the human 5-HTR2a gene to chromosome 13q14 and identification of two highly polymorphic linked markers suitable for association studies in psychiatric disorders. AB - The serotonergic system is known to play an important role in a number of psychiatric disorders. Indeed, treatments involving agents that have their pharmacological activities within this system are the mainstay of treatment for disorders such as schizophrenia. It is now widely accepted that many common psychiatric disorders have a familial or genetic component and as a result of this there has been an upsurge in interest in the 5-hydroxytryptamine (5-HT) pathways. A number of groups have attempted to establish whether polymorphism in particular proteins of the serotonergic system may form part of the genetic component of psychiatric disorders, including schizophrenia and anorexia nervosa. However, the data from these studies are conflicting and the problem is compounded by the lack of known polymorphic genetic markers mapping in close proximity to genes encoding proteins envolved directly or indirectly in 5-HT neurotransmission. In the current study, we have fine mapped the gene for 5-HTR2a by radiation hybrid mapping, and we report two new, highly linked, polymorphic markers that are suitable for linkage and association studies. PMID- 10464663 TI - Disease genes and chromosomes: disease maps of the human genome. Chromosome 21. PMID- 10464664 TI - Advances in the molecular genetics of congenital structural heart disease. AB - Molecular genetic analyses have generated significant advances in our understanding of congenital heart disease. Techniques of genetic mapping with polymorphic microsatellites and fluorescence in situ hybridization (FISH) have provided informative tools for localization and identification of disease genes. Some cardiovascular diseases have proven to result from single gene defects. Others relate to more complex etiologies involving several genes and their interactions. Elucidation of the molecular genetic etiologies of congenital heart disease prompts consideration of DNA testing for cardiac disorders. Future integration of these diagnostic modalities with improved treatments may ultimately decrease morbidity and mortality from congenital heart diseases. PMID- 10464665 TI - Meta-PCR: a novel method for creating chimeric DNA molecules and increasing the productivity of mutation scanning techniques. AB - Many mutation scanning techniques are capable of locating mutations in DNA fragments much larger than the average exon. We have developed a system called Meta-PCR that can maximize the length of sequence scanned by these techniques, improving their productivity and realizing their full potential. Meta-PCR is a simple, versatile, and powerful method for generating chimeric DNA molecules. Currently, up to five PCR amplifiable fragments can be combined to form a single linear amplimer. The Meta-PCR reaction is self-assembling and takes place in two coupled stages carried out in a single reaction vessel. The order of fragments is reproducible and determined by primer design. We have developed two Meta-PCR assays, one comprising exons 6-10 of the Neurofibromatosis type 2 (NF2) gene and the second exons 8-12 of the human mismatch repair gene, hMLH1. We verified by direct sequencing that the order and sequence of the component exons in the Meta PCR products is as predicted. Meta-PCR products from seven previously ascertained heterozygotes for NF2 mutations were directly sequenced. All seven mutations were clearly visible as mixed bases at the expected nucleotide, confirming that Meta PCR faithfully reproduces the original sample genotype. We have evaluated the downstream use of the NF2 Meta-PCR products in fluorescent solid-phase chemical cleavage of mismatches (CCM). Meta-PCR products from eleven NF2 mutant heterozygotes were screened retrospectively for piperidine cleavage after hydroxylamine or potassium permanganate modification of mismatched bases. Ten of the 11 mutants were detected by visible cleavage. One mutation predicted to be cleaved after potassium permanganate modification was not detected. However, we were able to attribute this false negative to a failure in the CCM method. Meta PCR is likely to be useful to clinical molecular diagnostic laboratories, helping them to fulfill demand for rapid and accurate screening for point mutations in large multi-exon genes. PMID- 10464666 TI - Prepregnancy testing for single-gene disorders by polar body analysis. AB - Preventive measures for single-gene disorders are currently based on carrier screening in pregnancy and prenatal diagnosis. Although this has been extremely effective for preventing new cases of common inherited conditions, the major limitation is still termination of 25% of wanted pregnancies following detection of affected fetuses. To overcome this important problem, we developed a method for prepregnancy genetic testing that involves DNA analysis of the first and second polar bodies, which are extruded during maturation and fertilization of oocytes. We offered this option to 28 couples at risk for having children with single-gene disorders. Fifty clinical cycles were performed from these patients for the following conditions: 20 for cystic fibrosis, 18 for thalassemia, 6 for sickle cell disease, 2 each for Gaucher disease and LCHAD (long-chain 3 hydroxyacyl-COA dehydrogenase deficiency), and 1 each for hemophilia B and phenylketonuria. Oocytes obtained from these patients using in vitro fertilization procedures (IVF) were tested by a sequential multiplex nested PCR analysis of the first and second polar body to detect the gene involved simultaneously with linked polymorphic markers. A total of 191 of 399 oocytes with predicted genotype were mutation free and preselected for fertilization and transfer. In all but three cycles, one to three unaffected embryos with predicted unaffected genotypes were transferred, resulting in 20 pregnancies, from which 19 healthy children have been born. The follow-up analysis of embryos resulting from oocytes with predicted affected genotype, confirmed the diagnosis in 97% of cases, demonstrating the reliability of prepregnancy diagnosis of single-gene defects by polar body analysis. PMID- 10464667 TI - The Genetic Knowledge Index: developing a standard measure of genetic knowledge. AB - This paper reports on the development of a unidimensional genetic knowledge index that has been tested and validated in a general population sample. The Index is intended to provide the basis for a standard measure of basic genetic knowledge that can be applied across diverse populations and research settings. The study group was composed of 330 European Americans selected randomly in the Louisville, KY, metropolitan area. The final version of the Genetic Knowledge Index (GKI) consisted of five items identified by principle components analysis, correlation coefficients, and the alpha measure of internal consistency. Construct validity of the GKI was determined by appropriate statistical correlations with educational attainment and attitudes toward genetic discrimination. The Index provides a numerical ranking of subjects' knowledge of practical genetics. Implications for research are discussed. PMID- 10464668 TI - A practical guide for the validation of genetic tests. AB - The unique aspects of genetic testing, such as the rarity of genetic disorders, lack of other tests for comparison, and heterogeneity and incomplete penetrance of some genetic diseases, make the validation of genetic tests a challenge in many instances. Although many organizations concerned with clinical laboratory testing and genetic testing in particular have acknowledged the need for validation of genetic tests prior to their introduction into routine practice, no guidelines exist on how to perform the validation of a genetic test. Given such difficulties faced by those involved in genetic testing, we present here a practical guide for the validation of genetic tests, compiled from the suggestions of several laboratorians involved in genetic testing and several officials involved in the oversight of laboratory testing in the United States. PMID- 10464669 TI - Screening Jews and genes: a consideration of the ethics of genetic screening within the Jewish community: challenges and responses. AB - Screening for genetic disorders, particularly Tay-Sachs Disease, has been traditionally welcome by the Jewish community. I review the history of genetic screening among Jews and the views from the Jewish tradition on the subject, and then discuss ethical challenges of screening and the impact of historical memories upon future acceptance of screening programs. Some rational principles to guide future design of genetic screening programs among Jews are proposed. PMID- 10464670 TI - Cystic fibrosis carrier screening practices in an ethnically diverse region: experience of the Genetic Network of the Empire State, Puerto Rico, and the U.S. Virgin Islands. AB - We surveyed clinical genetics centers to assess current cystic fibrosis (CF) screening practices with regard to clinical and laboratory aspects. The survey was developed by the CF committee of the Genetics Network of the Empire State, Puerto Rico, and the U.S. Virgin Islands (GENES) to gauge changes in trends following the April, 1997, NIH Consensus Statement recommending the offering of CF carrier screening to all pregnant patients. Thirty-five of 45 Centers (78%) returned the survey, which was mailed in June, 1998. Sixteen centers currently offer population-based screening, whereas 19 centers do not. Reasons cited for not offering testing included the low risk for CF in ethnic groups served, lack of data about test sensitivity in the populations served, and the absence of CF screening policies in the current standards of care. Approximately half (56%) of genetics centers that are offering testing altered their screening policy following the NIH Consensus statement, either by offering screening to patients of higher-risk ethnicities or by offering it to all patients. Less than half of the Centers that offer routine carrier screening offer screening to all patients regardless of ethnicity. This report is an initial step in documenting and understanding the current service practices regarding CF carrier testing in a diverse region. Our conclusions: (1) Screening practices vary widely among genetic centers in the region. (2) The decision to offer routine CF carrier screening is largely based on ethnicity of the patient population served. (3) Methods used to screen pregnant women and their partners in this part of the country reflect the diversity of models employed throughout the United States. (4) CF screening practices in the GENES region have changed significantly following the April, 1997, NIH consensus statement. PMID- 10464671 TI - Ashkenazi Jewish population frequency of the Bloom syndrome gene 2281 delta 6ins7 mutation. AB - Bloom syndrome is an autosomal recessive disorder characterized clinically by small size, sun-sensitive facial erythema, and immunodeficiency, and cytogenetically by increased chromosome breakage and sister chromatid exchange. Genomic instability renders Bloom syndrome patients at elevated risk for multiple cancers. Bloom syndrome occurs most commonly in the Ashkenazi Jewish population due to an apparent founder effect. The BLM gene on chromosome 15q26.1 was identified to encode a RecQ DNA helicase. Multiple mutations were identified, with Ashkenazi Jewish Bloom syndrome patients almost exclusively homozygous for a complex frameshift mutation (6-bp deletion/7-bp insertion at BLM nucleotide 2,281). This molecular genetic study seeks to verify the Ashkenazi Jewish carrier frequency of the BLM 2281 delta 6ins7 allele using semiautomated allele-specific oligonucleotide (ASO) analysis. Anonymized DNA samples from 1,016 Ashkenazi Jewish individuals and 307 non-Jewish individuals were screened. Ten Ashkenazi heterozygote carriers for the 2281 delta 6ins7 mutation were identified, giving a carrier frequency estimate of 0.98%, or approximately 1 carrier out of 102 individuals in the Ashkenazi Jewish population. These results are consistent with previous estimates, and combining our findings with the published molecular data collectively yields an Ashkenazi Jewish carrier frequency of approximately 1 in 104. Given its high population frequency and detection rate among Ashkenazi Jewish patients, the blmAsh mutation constitutes an appropriate addition to screening panels for Ashkenazi Jewish disease testing. PMID- 10464672 TI - BRCA1 screening in patients with a family history of breast or ovarian cancer. AB - Several studies have shown that the frequency of BRCA1 mutations in high-risk families differs widely between populations. In a recently published study from the Stockholm region, we found BRCA1 mutations in about 35% of the breast/ovarian families, but only in 1% of the families with site-specific breast cancer. To determine the frequency of BRCA1 mutations in families with a less increased risk for breast or ovarian cancer, a second study was performed. A total of 94 families with two and six families with only one affected member were included. Six mutations were found, all localized in exon 11, and five of them were previously known Swedish founder mutations. The mutation frequency was 6%, similar to the finding in families fulfilling the criteria for hereditary breast cancer (7%) that was disclosed in our first study. All families with a mutation had at least one individual with ovarian cancer. Thus, our study further implies that for a woman with breast cancer, a family history of ovarian cancer is far more important than a family history of breast cancer for predicting a BRCA1 mutation. Our results can be used to increase the specificity in selection of families for genetic testing. PMID- 10464673 TI - Mutations in the SLC3A1 gene in cystinuric patients: frequencies and identification of a novel mutation. AB - Cystinuria is a frequent autosomal recessive transport disorder characterized by defective renal resorption of cystine and other dibasic amino acids. Biochemically, three types of cystinuria can be defined. Here we present our results of screening for mutations in the SLC3A1 gene, which codes for a dibasic amino acid transporter protein and appears to be involved in the pathogenesis of cystinuria type I. Our study population consists of 5 Italian cystinuria type I patients and 10 cystinuric patients as yet unclassified as to clinical type. The latter were of different ethnic origin. In total, we found 13 point mutations and 8 genomic rearrangements in 15 cystinuric patients, i.e., our detection rate was 70% (23/30 chromosomes). Remarkably, in patients known to be suffering from cystinuria type I, the mutation detection rate was only 50%, whereas in patients unselected as to cystinuria type, we found 80% of mutations. Additionally, our results, as with those published in the literature, indicate a possible population specific distribution of mutations: Each of the 4 Greek patients analyzed here showed homozygosity for mutation T216M in exon 3. Analysis of a Yugoslavian patient showed homozygosity for a novel mutation, R365L, in exon 6 (nt1094G > T). Findings from molecular genetic studies, as well as physiological investigations, suggest that there are further genes that play a role in the etiology of cystinuria. Nevertheless, our results show that screening for mutations in the SLC3A1 gene can be a meaningful step toward molecular genetic diagnosis of cystinuria in patients without biochemical classification. As with cystic fibrosis, the finding of specific mutations in particular ethnic populations, suggest that the diagnostic approach should take into consideration a patient's ethnic origins. PMID- 10464674 TI - The 677C > T mutation in 5,10-methylenetetrahydrofolate reductase and colorectal cancer risk. AB - This paper examines whether there is a relationship between a common mutation in the methylenetetrahydrofolate reductase gene, MTHFR*val, and the risk of colorectal cancer, with or without lymph node metastases. MTHFR genotypes were ascertained from peripheral leukocyte samples obtained from 200 colorectal patients, including TMN stages I-VI, and from 460 healthy, unrelated adults without colorectal cancer, who served as controls. The frequency of homozygosity for the MTHFR*val/*val genotype among the colorectal cancer patients was lower (14.0%) than among controls (16.1%). The latter finding results in an estimated MTHFR*val allele frequency of 0.41. The MTHFR*val allele (677C > T) reduces colorectal risk slightly [odds ratio (OR), 0.87]. However, there was a significantly higher incidence of metastatic lymph nodes per case in MTHFR*val/*val patients, when compared with MTHFR*ala/*ala controls (6.9 +/- 1.55 vs. 3.7 +/- 0.57, p = 0.003). These results suggest that the MTHFR genotype might be of prognostic significance in colorectal carcinoma. PMID- 10464675 TI - HER-2/neu oncogene amplification in cervical cancer studied by fluorescent in situ hybridization. AB - Oncogene amplification, such as HER-2/neu (C-erbB-2), is a manifestation of genetic instability often associated with the genesis and progression of cancer, including cervical cancer. Oncogene overexpression is traditionally studied using immunohistochemistry. We previously reported studies of oncogene amplification in breast cancer using fluorescent in situ hybridization (FISH), where the data support the hypothesis that HER-2/neu is a prognostic marker of poor outcome. To explore further the possible significance of HER-2/neu oncogene amplification in cervical cancer, we conducted a pilot study of 24 cervical cancer cases. The HER 2/neu FISH probe (Vysis, Inc., Downers Grove, IL) was used to measure gene amplification, with a chromosome 17 centromeric probe as an internal control. Out of 24 cases studied, 23 were informative. Of the 23 informative cases, 2 (8.7%) were found to be amplified. The rest (21 out of 23 or 91.3%) were nonamplified. Both amplified cases were invasive adenocarcinoma. Although the sample size of this pilot study may be somewhat small, the data obtained so far clearly demonstrated that detection of oncogene amplification in cervical cancer is not only feasible but is very sensitive, and suggest that further exploration using a larger sample size may be warranted. PMID- 10464676 TI - Disease genes and chromosomes: disease maps of the human genome. Chromosome 16. PMID- 10464677 TI - [Pulsed laser in scar treatment]. AB - Pulsed laser systems such as pulsed CO2, Q-switched Er:YAG, ruby and alexandrite lasers provide good therapeutic options in therapy of scars and keloids. The use of these laser systems is compared to non-surgical and surgical methods. PMID- 10464678 TI - [Clinical and epidemiological data of patients with malignant melanoma from the Munich Tumor Center 1977-1997]. AB - Since 1997, data of patients with malignant melanomas have been systematically documented in the tumor registry of the Tumor Center Munich. Analysis of data of 8071 patients revealed that tumor thickness has steadily declined over the years. While in 1977 the median tumor thickness was 1.45 mm, it is now 0.75 mm. This has been followed by a significant improvement in overall survival. Males and older patients tend to have thicker melanomas than females and younger patients. There has been a relative increase of melanomas of the trunk. At diagnosis, 95% of patients had local disease. Of these patients, 18.3% developed metastastes. At least two-thirds of these patients had progression at the primary tumor site or the regional lymph nodes, both of which can be assessed by clinical or ultrasound examinations. Overall survival of patients with thin melanomas is excellent and does not differ substantially from the overall survival of the general population comparable in sex and age. PMID- 10464679 TI - [How can Romberg disease be evaluated by means of MRI?]. AB - Romberg disease, also known as progressive facial hemiatrophy (PFH), is an uncommon disease with progressive facial asymmetry as the major symptom. It is a atrophic process of the subcutaneous fatty tissue whose etiology is unknown. Skin, muscles and bones can be secondarily afflicted. With MRI evaluation we were able to show in 14 patients with PFH thinner subcutaneous fat, as well as flattening of the dermis in 12 of the 14. We divided the patients into three stages by the means of the MRI findings and found tendency towards concurrence between the MRI stage and the clinical stage. PMID- 10464680 TI - [Pachyonychia congenita. Keratin gene mutations with pleiotropic effect]. AB - Pachyonychia congenita (PC) comprises a heterogeneous group of autosomal dominantly inherited conditions showing characteristic nail thickening and associated signs such as palmoplantar keratoderma, follicular keratoses, and mucosal leukokeratoses. Less frequently epidermal cysts, hairshaft abnormalities, natal teeth and laryngeal involvement may be seen. Phenotypically and genetically two major forms of PC are recognized, pachyonychia congenita Jadassohn Lewandowsky/PC type I (Medelian inheritance in man-MIM-167200) and pachyonychia congenita Jackson-Lawler/PC type II (MIM 167210). Both conditions show nail deformities, focal palmoplantar keratoderma, and follicular hyperkeratoses. Diagnostically relevant are leukokeratoses of the oral mucosa in patients with PC type I. In contrast individuals affected with PC type II show premature dentition and multiple pilosebaceous cysts predominantly affecting the upper trunk. The latter closely resemble eruptive vellus hair cysts and steatocystoma multiplex. By mutational analysis keratin K6a and K16 gene mutations have been detected in patients with PC type I, and keratin K6b and K17 gene mutations have been shown to be the underlying genetic defect in patients with PC type II. PMID- 10464681 TI - [Tubero-pustular demodicosis]. AB - A 38-year-old female patient suddenly developed an unusual tuberous, pustular tumor on her chin. On the basis of clinical pattern, histological and microbiological investigations the diagnosis of demodicosis was established. Histological investigation revealed follicular cysts and a chronic granulomatous perifolliculitis with many of Demodex folliculorum. A large number of mites could be also identified by microscopy of smears from pustules. No cellular or humoral immunological defects, tumours nor systemic disorders were found. After oral therapy with steroids and metronidazole, the lesions improved rapidly. PMID- 10464682 TI - [Papular-purpuric gloves and socks syndrome]. AB - In 1990, we first described the papular-purpuric "gloves and socks" syndrome (PPGSS). It is characterized by an itching erythema and edema of the hands and feet together with oral mucosal lesions and fever. The skin lesions subsequently become purpuric and resolve spontaneously after a few days. In more than half of the patients, an acute parvovirus B 19 infection has been proven. In a few cases of PPGSS, other viral infections have been found, for example hepatitis B and measles. We report on a patient with PPGSS and review the literature. PMID- 10464683 TI - [Acrodermatitis enteropathica-like changes in a patient with parenteral nutrition]. AB - A patient on long term parenteral nutrition developed acrodermatitis enteropathica-like skin changes. Just as in the autosomal recessive disorder, acrodermatitis enteropathica, the lesions in our patient were caused by zinc deficiency. Replacement therapy with zinc led to rapid improvement. PMID- 10464684 TI - [Recurrent episodes of ulcerative gingivostomatitis associated with cyclic neutropenia]. AB - Cyclic neutropenia is a rare hematologic disorder of unknown origin. Periodic decreases in blood leukocytes with relative neutropenia and lymphocytosis are typical. These episodes appear in regular intervals of 15-35 days and last three to seven days. They are associated with physical weakness, fever, septic complications and necrosis of mucous membranes. Between the episodes, the patients are in complete clinical and laboratory remission. We report the case of a 34 year-old patient suffering from cyclic neutropenia with ulcerous lesions of the oral mucosa. PMID- 10464685 TI - [Radiation-induced morphea]. AB - Although the aetiology of morphea often remains unknown, various precipitating causes have been identified. Morphea is a rare development following irradiation. We observed a circumscribed scleroderma 14 years after x-ray-radiotherapy following surgery of breast cancer. Furthermore multiple weeping papules occurred within the radiation area. A relapse of the breast cancer was ruled out by histological examination which displayed a transepidermal elimination of acid mucin. The irradiation had obviously induced different reactions of the dermal fibroblasts (radiodermatitis, morphea, mucin production). PMID- 10464686 TI - [UVA dose during treatment at the Dead Sea]. PMID- 10464687 TI - [Comment on the report "Successful treatment of Lyell's syndrome with intravenous immunoglobulins". ("Gehort-gelesen-nachgefragt") Hautarzt (1998) 49:950-951]. PMID- 10464688 TI - [Exanthema in childhood. I: Exanthema induced by viral infections]. PMID- 10464689 TI - A fluid--structure interaction finite element analysis of pulsatile blood flow through a compliant stenotic artery. AB - A new model is used to analyze the fully coupled problem of pulsatile blood flow through a compliant, axisymmetric stenotic artery using the finite element method. The model uses large displacement and large strain theory for the solid, and the full Navier-Stokes equations for the fluid. The effect of increasing area reduction on fluid dynamic and structural stresses is presented. Results show that pressure drop, peak wall shear stress, and maximum principal stress in the lesion all increase dramatically as the area reduction in the stenosis is increased from 51 to 89 percent. Further reductions in stenosis cross-sectional area, however, produce relatively little additional change in these parameters due to a concomitant reduction in flow rate caused by the losses in the constriction. Inner wall hoop stretch amplitude just distal to the stenosis also increases with increasing stenosis severity, as downstream pressures are reduced to a physiological minimum. The contraction of the artery distal to the stenosis generates a significant compressive stress on the downstream shoulder of the lesion. Dynamic narrowing of the stenosis is also seen, further augmenting area constriction at times of peak flow. Pressure drop results are found to compare well to an experimentally based theoretical curve, despite the assumption of laminar flow. PMID- 10464690 TI - Velocity and wall shear stress patterns in the human right coronary artery. AB - Blood flow dynamics in the human right coronary artery have not been adequately quantified despite the clinical significance of coronary atherosclerosis. In this study, a technique was developed to construct a rigid flow model from a cast of a human right coronary artery. A laser photochromic method was used to characterize the velocity and wall shear stress patterns. The flow conditions include steady flow at Reynolds numbers of 500 and 1000 as well as unsteady flow with Womersley parameter and peak Reynolds number of 1.82 and 750, respectively. Characterization of the three-dimensional geometry of the artery revealed that the largest spatial variation in curvature occurred within the almost branch-free proximal region, with the greatest curvature existing along the acute margin of the heart. In the proximal segment, high shear stresses were observed on the outer wall and lower, but not negative, stresses along the inner wall. Low shear stress on the inner wall may be related to the preferential localization of atherosclerosis in the proximal segment of the right coronary artery. However, it is possible that the large difference between the outer and inner wall shear stresses may also be involved. PMID- 10464691 TI - The effects of cross-fiber deformation on axial fiber stress in myocardium. AB - We incorporated a three-dimensional generalization of the Huxley cross-bridge theory in a finite element model of ventricular mechanics to examine the effect of nonaxial deformations on active stress in myocardium. According to this new theory, which assumes that macroscopic tissue deformations are transmitted to the myofilament lattice, lateral myofilament spacing affects the axial fiber stress. We calculated stresses and deformations at end-systole under the assumption of strictly isometric conditions. Our results suggest that at the end of ejection, nonaxial deformations may significantly reduce active axial fiber stress in the inner half of the wall of the normal left ventricle (18-35 percent at endocardium, depending on location with respect to apex and base). Moreover, this effect is greater in the case of a compliant ischemic region produced by occlusion of the left anterior descending or circumflex coronary artery (26-54 percent at endocardium). On the other hand, stiffening of the remote and ischemic regions (in the case of a two-week-old infarct) lessens the effect of nonaxial deformation on active stress at all locations (9-32 percent endocardial reductions). These calculated effects are sufficiently large to suggest that the influence of nonaxial deformation on active fiber stress may be important, and should be considered in future studies of cardiac mechanics. PMID- 10464692 TI - Quasi-Linear Viscoelastic theory applied to internal shearing of porcine aortic valve leaflets. AB - The elements of Quasi-Linear Viscoelastic (QLV) theory have been applied to model the internal shear mechanics of fresh and glutaraldehyde-fixed porcine aortic valve leaflets. A novel function estimation method was used to extract the material functions from experimental shear data obtained at one strain rate, and the model was used to predict the material response at different strain rates. In general, experiments and predictions were in good agreement, the larger discrepancies being in the prediction of peak stresses and hysteresis in cyclic shear. In shear, fixed tissues are stiffer (mean initial shear modulus, 13 kPa versus 427 Pa), take longer to relax to steady state (mean tau 2 4,736 s versus 1,764 s) with a slower initial relaxation rate (mean magnitude of G(0), 1 s-1 versus 5 s-1), and relax to a lesser extent than fresh tissues (mean percentage stress remaining after relaxation, 60 versus 45 percent). All differences were significant at p = 0.04 or less, except for the initial relaxation slope. We conclude that shear experiments can complement traditional tensile and biaxial experiments toward providing a complete mechanical description of soft biomaterials, particularly when evaluating alternative chemical fixation techniques. PMID- 10464693 TI - Calibration and application of an intra-articular force transducer for the measurement of patellar tendon graft forces: an in situ evaluation. AB - The objective of this study was to evaluate two calibration methods for the "Arthroscopically Implantable Force Probe" (AIFP) that are potentially suitable for in vivo use: (1) a direct, experimentally based method performed by applying a tensile load directly to the graft after it is harvested but prior to implantation (the "pre-implantation" technique), and (2) an indirect method that utilizes cadaver-based analytical expressions to transform the AIFP output versus anterior shear load relationship, which may be established in vivo, to resultant graft load (the "post-implantation" technique). The AIFP outputs during anterior shear loading of the knee joint using these two calibration methods were compared directly to graft force measurements using a ligament cutting protocol and a 6 DOF load cell. The mean percent error (actual-measured)/(actual)* 100) associated with the pre-implantation calibration ranged between 85 and 175 percent, and was dependent on the knee flexion angle tested. The percent error associated with the post-implantation technique was evaluated in two load ranges: loads less than 40 N, and loads greater than 40 N. For graft force values greater than 40 N, the mean percent errors inherent to the post-implantation calibration method ranged between 20 and 29 percent, depending on the knee flexion angle tested. Below 40 N, these errors were substantially greater. Of the two calibration methods evaluated, the post-implantation approach provided a better estimate of the ACL graft force than the pre-implantation technique. However, the errors for the post implantation approach were still high and suggested that caution should be employed when using implantable force probes for in vivo measurement of ACL graft forces. PMID- 10464694 TI - Quantification of structural loading during off-road cycling. AB - To provide data for fatigue life prediction and testing of structural components in off-road bicycles, the objective of the research described herein was to quantify the loads input to an off-road bicycle as a result of surface-induced loads. A fully instrumented test bicycle was equipped with dynamometers at the pedals, handlebars, and hubs to measure all in-plane structural loads acting through points of contact between the bicycle and both the rider and the ground. A portable data acquisition system carried by the standing rider allowed, for the first time, this loading information to be collected during extended off-road testing. In all, seven experienced riders rode a downhill trial test section with the test bicycle in both front-suspension and full-suspension configurations. The load histories were used quantitatively to describe the load components through the computation of means, standard deviations, amplitude probability density functions, and power spectral density functions. For the standing position, the coefficients of variation for the load components normal to the ground were greater than 1.2 for handlebar forces and 0.3 and 0.5-0.6 for the pedal and hub forces, respectively. Thus, the relative contribution of the dynamic loading was much greater than the static loading at the handlebars but less so at the pedals and hubs. As indicated by the rainflow count, high amplitude loading was developed approaching 3 and 5 times the weight of the test subjects at the front and rear wheels, respectively. The power spectral densities showed that energy was concentrated in the band 0-50 Hz. Through stress computations and knowledge of material properties, the data can be used analytically to predict the fatigue life of important structural components such as those for steering. The data can also be used to develop a fatigue testing protocol for verifying analytical predictions of fatigue life. PMID- 10464695 TI - On the kinematic modeling and the parameter estimation of the human knee joint. AB - This paper presents some results on the modeling and the parameter estimation of the human knee joint. Based on the geometric characteristics of the femur condyle and the tibia plateau, a part of femoro-tibial joint model includes an involute on-plane submodel. Data recorded by camera type device are used to analyze the kinematic characteristics of the knee joint and to estimate the corresponding submodel parameters. Experimental results are presented and the model is further validated. PMID- 10464696 TI - A cellular solid criterion for predicting the axial-shear failure properties of bovine trabecular bone. AB - In a long-term effort to develop a complete multi-axial failure criterion for human trabecular bone, the overall goal of this study was to compare the ability of a simple cellular solid mechanistic criterion versus the Tsai-Wu, Principal Strain, and von Mises phenomenological criteria--all normalized to minimize effects of interspecimen heterogeneity of strength--to predict the on-axis axial shear failure properties of bovine trabecular bone. The Cellular Solid criterion that was developed here assumed that vertical trabeculae failed due to a linear superposition of axial compression/tension and bending stresses, induced by the apparent level axial and shear loading, respectively. Twenty-seven bovine tibial trabecular bone specimens were destructively tested on-axis without end artifacts, loaded either in combined tension-torsion (n = 10), compression torsion (n = 11), or uniaxially (n = 6). For compression-shear, the mean (+/- S.D.) percentage errors between measured values and criterion predictions were 7.7 +/- 12.6 percent, 19.7 +/- 23.2 percent, 22.8 +/- 18.9 percent, and 82.4 +/- 64.5 percent for the Cellular Solid, Tsai-Wu, Principal Strain, and von Mises criteria, respectively; corresponding mean errors for tension-shear were -5.2 +/- 11.8 percent, 14.3 +/- 12.5 percent, 6.9 +/- 7.6 percent, and 57.7 +/- 46.3 percent. Statistical analysis indicated that the Cellular Solid criterion was the best performer for compression-shear, and performed as well as the Principal Strain criterion for tension-shear. These data should substantially improve the ability to predict axial-shear failure of dense trabecular bone. More importantly, the results firmly establish the importance of cellular solid analysis for understanding and predicting the multiaxial failure behavior of trabecular bone. PMID- 10464697 TI - Low-frequency acoustic sweep monitoring of bone integrity and osteoporosis. AB - We developed a noninvasive method to evaluate bone structural integrity. It is based on the measurement of the dynamic characteristics of the bone using sweeping sound excitation in the range of acoustic frequencies. The Quality Factor (a measure of material damping) has been used as an indicator of the tendency of the bone to fracture. Results of animal studies have supported this hypothesis since linear correlations were observed between bone density, quality factor, and impact strength. A vibration excitation in the form of an acoustic sweep signal is applied to a bone to measure the quality factor. Rat bones were tested, obtained from animals with osteoporosis age-dependent (tested in vitro) or ovariectomy-induced (tested in vivo), and compared with bones of healthy (control) rats. The change in damping was, on average, equal or greater to the change in density. Moreover, excellent correlation of the quality factor was obtained with bone fracture energy measured with an impact test. During a vibration cycle, the changing strain results in temperature changes due to the reciprocity of temperature and strain. Nonreversible conduction of heat due to the unequal temperature change results in entropy production that is enhanced due to the stress concentration about the voids associated with bone porosity. Damping is a measure of the production of entropy. Its measure, the quality factor, represents a potentially useful tool for monitoring bone integrity, which is deteriorating in diseases characterized by disruption of the trabecular architecture, such as osteoporosis. A computational model yielded results that are in good correlation with the experimental results. PMID- 10464698 TI - Carcinoid syndrome: a statistical evaluation of 748 reported cases. AB - No statistical evaluation of patients with carcinoid syndrome in a reliable number of cases has been available in the past 35 years. To update our knowledge about the syndrome, we have evaluated from various clinicopathologic viewpoints a large series of patients with the syndrome reported up to date. The data of 748 patients with the syndrome were collected from 8876 carcinoid patients reported in the literature and analyzed by the Gut-Pancreatic Endocrinoma Analyzing System (the Niigata Registry). The results are summarized as follows. 1) The patients with the syndrome had a tendency to be older than those without it. 2) The incidence of the syndrome was 8.4% of 8876 carcinoid patients. 3) Serotonin activities were extremely high in patients with the syndrome as compared to those without it (91.7% versus 26.6%). 4) The rate of metastases was higher in patients with the syndrome than in those without it (84.8% versus 29.2%), and higher in the liver than in lymph nodes among patients with the syndrome (73.4% versus 37.4%). 5) Flushing and carcinoid heart as most specific clinical manifestations of the syndrome were recorded at 78.3% and 17.4%, respectively. 6) The 5-year survival rate after resection of primary lesions was 76.0% of 304 patients with the syndrome, lower in patients with digestive carcinoids than in those with extradigestive lesions (67.2% versus 88.7%). It is expected that the results obtained in the present evaluation on patients with carcinoid syndrome will provide investigators active in this specialized field with useful and extensive information for their future activities. PMID- 10464699 TI - Prognosis of patients with stage IIIb-IVa squamous cell carcinoma of the cervix following intra-arterial neoadjuvant chemotherapy. AB - Our purpose was to determine the long-term prognosis in patients with stage IIIb IVa squamous cell carcinoma of the cervix who were treated with intra-arterial neoadjuvant chemotherapy (NAC), and to analyze factors related to prognostic value. We assessed the disease-free survival of 21 patients with FIGO stage IIIb IVa squamous cell carcinoma of the cervix treated with intra-arterial NAC (cisplatin 70 mg/m2 and peplomycin sulfate 30 mg/m2) followed by irradiation therapy. Before chemotherapy, five factors (age, clinical stage, histologic type, parametrial involvement and serum level of SCC) were evaluated for their correlation with disease-free survival. The absolute 5-year disease-free survival rate for the entire group was 52%, with a median follow-up time in disease-free survivors of 87 months. Among the prognostic factors, age was the only one to be significantly correlated with disease-free survival; older (> 60 years) patients showed a significantly better disease-free survival rate than did younger (< 60 years) patients (p < 0.05, log-rank test). The absolute 5-year disease-free survival rate was 69% for older patients and 25% for younger patients. Univariate Cox's proportional hazard model also demonstrated that age was a significant prognostic factor as a continuous variable (p < 0.01). Intra-arterial NAC thus appeared to be effective in treating older patients with stage IIIb-IVa squamous cell carcinoma of the cervix. PMID- 10464700 TI - Total thyroidectomy and synchronous video assisted lung metastasectomy in differentiated thyroid carcinoma. PMID- 10464702 TI - Can traditional epidemiology detect cancer risks caused by occupational exposure to pesticides? AB - In order to investigate the possible relationship between cancer and occupational exposure to pesticides, we reviewed the latest literature of the epidemiological studies in this area coming to the conclusion that, while several studies indicate a link between certain pesticides and certain tumors, this information is still insufficient, and further research on the health consequences of exposure to pesticides is needed. Moreover, provided there is a risk, it is often too limited to be detected by available epidemiological techniques. Therefore, in addition to the epidemiological studies, the development of new biology, gene technology and medical biotechnology methods may significantly enhance the specificity of the epidemiological studies. Thus, the fusion of molecular biology and epidemiology into molecular epidemiology may provide more specific methods for monitoring the occupational dependent carcinogenic risk of individuals and groups. PMID- 10464701 TI - The integrated treatment of peritoneal carcinomatosis. A preliminary experience. AB - Some low-grade malignant tumors arising in the abdomen, lack of infiltrative attitude and "redistribute" on the peritoneum with no extraregional spreading. In this cases the complete tumor cytoreduction followed by intra- or postoperative regional chemotherapy has curative intent. Peritonectomy is the complete removal of all the parietal peritoneum and the visceral peritoneum involved by disease. After peritonectomy hyperthermic antiblastic perfusion is carried out throughout the abdomino-pelvic cavity for 60 minutes, at a temperature of 41.5 degrees C, with mitomycin C (3.3 mg/m2/Lt of perfusate) and cisplatin (25 mg/m2/Lt) (appendicular or colorectal primary), or cisplatin alone is (ovarian primary). Alternatively the immediate postoperative regional chemotherapy is performed with 5-fluorouracil (13.5 mg/Kg) and Lederfolin (125 mg/m2) (colic or appendicular tumor) or cisplatin (25 ng/m2) (ovarian tumor), each day for 5 days. Twenty patients affected by extensive peritoneal carcinomatosis (12 ovarian, 5 colonic, 1 appendicular, 1 mesothelial and 1 gastric primary) were submitted to peritonectomy with no residual macroscopic disease in all cases except three. Six patients were treated with intraoperative intra-abdominal hyperthermic antiblastic perfusion, while immediate postoperative intra-abdominal chemotherapy was given in 4 patients and systemic chemotherapy in other 5. Hospital mortality was 20%. At a mean follow-up of 11 months 14 patients are alive, 11 without disease and the median overall survival is 10.2 months. The curative potential of the combined therapeutic approach seems high in patients with peritoneal carcinomatosis from ovarian or colorectal primary not responding to systemic chemotherapy. Selection criteria of patients can strictly affect the surgical risk and the treatment has to be reserved for controlled clinical trials. PMID- 10464703 TI - The effect of pretreatment with docetaxel (taxotere; RP 56976) on irradiated subcutaneous MA 16/C murine tumors. AB - Docetaxel (taxotere; RP 56976) is twice as potent as taxol in the inhibition of microtubule depolymerisation and accumulates cells in their G2/M-phase, which is a highly sensitive phase of the cell cycle. The present study was designed to test the hypothesis that this property may lead to an enhanced effect of ionizing radiation on tumors. In a first experiment subcutaneous MA 16/C murine tumors, which are highly responsive to docetaxel, were treated intravenously with 45 mg/kg docetaxel and 6 hours later irradiated with 500 cGy (day 1). The pretreatment time of 6 hours resulted in a maximum mitotic index at the time of irradiation. On day 3 and 5, an additional 500 cGy was administered, without pretreatment with docetaxel. In a second experiment the radiation dose was given similarly, but pretreated with 15 mg/kg docetaxel. In both experiments, a prolonged increase of lifespan (%ILS) and a tumor growth delay (Td) was found among docetaxel treated mice and combinedly treated animals. Though a radiation dose of 1500 cGy on its own had no antitumoral effect on MA 16/C tumors, there was a marked tendency (but not statistically significant) to a better effect with the combined therapy in comparison with docetaxel alone (in experiment 1). Further studies with more radiosensitive tumors are warranted to determine the putative role of M-phase arrest on the radiosensitizing properties of docetaxel. PMID- 10464704 TI - Enhanced antitumor immune responses of IL-2 gene-modified tumor vaccine by combination with IL-1 and low dose cyclophosphamide. AB - To enhance the antitumor immunity induced by IL-2 gene-modified tumor vaccine, we proposed a combined protocol to treat tumor-bearing mice using IL-2 gene-modified tumor vaccine in combination with IL-1 and low-dose Cyclophosphamide(Cy). After treatment with IL-2 gene-modified B16 melanoma cell vaccine alone, the pulmonary metastases of tumor-bearing mice were reduced and their survival time was prolonged. The anti-metastases effect was improved when the vaccine was used in combination with IL-1 or low-dose Cy. The best therapeutic effect was achieved when the IL-2 gene-modified vaccine was combined with IL-1 and low-dose Cy. The cytotoxicity of the splenic CTL, NK, and the levels of IL-2, TNF secreted by splenocytes increased after tumor-bearing mice were treated with the IL-2 gene modified tumor vaccine. The above antitumor immune functions were augmented more significantly when IL-1, low-dose Cy were used in combination with IL-2 genemodified tumor vaccine. These results demonstrated that the IL-2 gene modified vaccine could exert more potent anti-metastases effects when it is combined with IL-1 or/and low-dose Cy by activating the specific and non-specific antitumor immune responses more effectively. PMID- 10464705 TI - Thermosensitivity, incidence of apoptosis and accumulations of hsp72 and p53 proteins of murine L cells in wild type status of p53 gene. AB - Murine L cells showed markedly high lethal thermosensitivity. Survivals from fractionated heating at 44 degrees C with variety of interval time at 37 degrees C (44 degrees C for 10 min--variety of interval time at 37 degrees C-44 degrees C for 10 min) increased markedly in accordance with elongation of the internal time; i.e. survival fraction of 0.9% from 44 degrees C for 20 min alone without the interval time to those of 25% from the fractionated heating with interval time at 37 degrees C for 3-10 hrs. Incidence of apoptosis of the L cells from heating at 44 degrees C for 6.5 min (LD50) increased from 7% immediately after the heating to 30% 6-12 hrs after the post-incubation time at 37 degrees C. Accumulation of both hsp72 and p53 proteins markedly increased after a heating at 44 degrees C for 10 min alone in accordance with elongation of post-incubation time at 37 degrees C, representing a peak 6 hrs after the post incubation. Status of p53 gene in L cells were determined with Reverse Transcription-Polymerase Chain Reaction-Single Strand Conformational Polymorphism (RT-PCR-SSCP), i.e. wild type. PMID- 10464706 TI - Efficient inducation of local and systemic antitumor immune response by liposome mediated intratumoral co-transfer of interleukin-2 gene and interleukin-6 gene. AB - Interleukin 2 (IL-2) expressing plasmid and interleukin 6 (IL-6)-expressing plasmid were encapsulated in liposome and administrated intratumoraly into tumor bearing mice 4 days after subcutaneous inoculation of B16F10 melanoma cells. The results showed that treatment of tumor-bearing mice with IL-2 gene or IL-6 gene transfer inhibited the growth of subcutaneous tumor and prolonged the survival of tumor-bearing mice significantly when compared with the treatment of PBS or control gene transfer mediated by liposome (P < 0.01). Combined transfer of IL-2 gene and IL-6 gene was found to elicit inhibitory effects on the growth of B16F10 tumor more significantly and prolonged the survival period of tumor-bearing mice more obviously. We investigated the local immunity in tumor microenvironment and found that IL-2 and IL-6 gene transfer could significantly increase the expression of lymphocyte function-associated antigen-1 on tumor infiltrating lymphocytes (TIL) and MHC-I molecule on tumor cells freshly isolated from the tumor mass. The NK and CTL activity of TIL increased markedly after the combined transfer of these two cytokine genes. We also observed the systemic antitumor immune response in the tumor-bearing mice and demonstrated that NK and CTL activity of splenocytes and the production of IL-2, tumor necrosis factor and interferon-gamma from splenocytes increased obviously in mice after the combined transfer of IL-2 and IL-6 gene. In conclusion, local and systemic antitumor immunity of the tumor-bearing host could be induced efficiently after the combined gene transfer. The enhanced specific and non-specific antitumor immunity might be responsible for the more potent antitumor effects of the combined gene therapy. PMID- 10464707 TI - Graft versus host disease in autologous stem cell transplantation. AB - Relapse remains the major cause of mortality in haematological malignancies treated with autologous stem cell transplantation (ASCT). Graft versus tumour reaction (GVT) associated to autologous graft versus host disease (GVDH) may contribute to eliminate minimal residual disease (MRD) after ASCT. Eighty patients with several diagnostics were submitted to ASCT. After stem cell infusion, patients randomised in 4 groups. Groups were treated as follows: Group A received either a IFN (alpha Interferon--1,000,000 U/d), Cyclosporine A (CSA--1 mg/-kg/d intravencus) for 28 days, and granulocyte-macrophage colony stimulating factor (GM-CSF-250/m2/d) until engraftment; B: CSA (same dose and way) and GM CSF; C: CSA (1 mg/kg/d orally) and GM-CSF and D: only GM-CSF. Patients were inspected daily and if skin rash was detected, a skin biopsy was obtained at that moment, otherwise biopsies were obtained at day 21 after ASCT. GVHD was positive in 23 patients (13 from group A and 10 from group B). All cases were grades I and II. A majority of CD4+ T lymphocytes was seen in skin infiltrates. No significant differences were seen in WBC and platelets engraftment times, antibiotic administration or hospitalisation days required among the four groups. With a median follow up of 18 months, there were no differences in disease free survival (DFS) or overall survival (OS) between the patients who developed GVHD and the others. However, considering that myeloma cells do not express antigen MCH II, which is necessary for GVT effect, we excluded patients with multiple myeloma (MM) from survival analysis, thus obtaining a significant difference in OS results between patients who developed GVHD and those in whom this reaction was not observed (81% vs 58% p:0.05). We conclude that pharmacological induction of GVHD in ASCT is possible with CSA administration (1 mg/kg/d i.v.). Development of GVHD showed a better outcome for patients in our study except for those patients with MM. This results must be confirmed by a longer follow up of our patients and further studies. PMID- 10464708 TI - Age as a risk factor in cervicofacial reconstruction. AB - The Authors report their experience from January 1980 to January 1998 with 392 reconstructive flaps for the oncological reconstruction of the cervicofacial district. One hundred and forty-two were conventional flaps, 187 were myocutaneous or pedicled muscular ones and 63 were microvascular. Ninety percent of the patients had Stage IV disease, 80% were tumors involving the oral cavity structures, 40% of the patients had received preoperative radiotherapy; 35% of the cases were recurrences. The complications related to the surgery were evaluated for each type of flap, comparing them to the same flaps employed in patients > 70. Complications were divided into major and minor that were observed at an overall rate of 22%. With the use of conventional flaps, there was an 11.3% rate of minor complications and a 5.6% rate of major ones. Examining the 187 myocutaneous or pedicled muscular flaps, 21% of the overall 27% were minor and 6% were major complications. Of the remaining 63 free flaps, minor complications were observed in 4.7% and major complications in 14.2% of cases. Thirty-four of the 392 flaps, 24 of which were myocutaneous or pedicled muscular and 10 free flaps, were utilized in elderly patients and compared with the 216 of the same type, in patients < 70. A comparative analysis shows that there was a major complication rate of 11.7% in the flaps employed on the elderly patients as opposed to 7.9% for those employed in the younger patients. In terms of minor complications, a 20.5% complication rate was observed for those > 70 as opposed to 16.7% for patients < 70. A more detailed analysis of these data, enabled to postulate that the smaller group of flaps used in elderly patients is statistically influenced by the "dilution" of the complication rate in favour of the larger group of younger patients. Furthermore, by appropriately correcting the risk factors due to concomitant diseases that were not related to surgery in the older patients, a realignment of the results may be seen. Therefore, a careful preoperative study must be carried out in the elderly patients with cervicofacial tumors who are eligible for surgery to establish and possibly treat the concomitant disease responsible for the increased peri- and postoperative morbidity. In conclusion, complications in elderly patients are correlated to the state of co-morbidity and neither to age nor to the duration of the operation. Responsiveness may therefore be obtained also in elderly patients employing sophisticated techniques such as microvascular flaps. PMID- 10464709 TI - Relationship between intrathyroid calcifications and thyroglobulin in endemic goiter. AB - Intrathyroid calcifications represent a common finding within simple or nodular goiters, but, as far as they can be found also inside papillary and medullary thyroid carcinomas, an ultrasonographic detection of intrathyroid calcifications stands as a different diagnosis problem. We have been looking for the presence of parameters associated with thyroid calcifications in patients affected by simple or nodular goiter, either sporadic or endemic. We studied 284 euthyroid subjects, 250 females, ageing from 24 to 90 years, affected by a simple goiter, in the 9.51% of the cases, and by a nodular goiter in the remaining part. 69.37% of the patients came from an endemic goiter area, while the others were affected by sporadic goiter. We tested fT3, fT4, TSH, hTG, Ab-TG, Ab-TPO and performed an ultrasonography in all the subjects, 57.75% of patients shown intrathyroid calcifications in the 57.75% of them. We applied a multistep discriminant analysis taking for the presence/absence of calcifications as dependent variable and we tried to find which variable, by itself or in combination with others, could foretell its presence. We also created a new variable (TG1) to differentiate normal from supraphysiologic concentrations of hTG (< 60 ng/ml). The variable with the highest significance F originated from endemic goiter area (F = 96.36), followed by TG1 (F = 24.46) and age (F = 10.61). On the contrary hTG did not relate to calcifications, due to non-proportionally direct relationship between these two parameters, afterwards we used the multistep logistic regression that gave overlapping significances. This means that supraphysiologic hTG rates are sufficient to predict the possible presence of intrathyroid calcifications. In conclusion, as far as a follicular hyperstimulation can be assumed, especially if long-lasting, the presence of intrathyroid calcifications should rise a clinical suspect toward an old goiter rather than a neoplastic lesion. PMID- 10464710 TI - Medical decision making for melanoma of the glans penis. AB - This case report describes a rare presentation of penile melanoma in which 3 successive primaries arose and were operated from an area of melanosis on the glans penis and prepuce. One of the major factors accounting for the poor prognosis of this patient was the long delay in presentation. This was largely due to the patient's reluctance because of the site of the disease. When diffuse melanotic areas are present in the genital region, in particular given the reluctance of patients with skin lesions in this region to present, the index of suspicion should be high with respect to the risk of transformation and an aggressive follow-up policy should be advocated. Treatment guidelines should not significantly differ from the usual approach of cutaneous melanoma. PMID- 10464711 TI - Prognosis of a family with familial medullary thyroid carcinoma. PMID- 10464712 TI - Differential expression of transforming growth factor beta 1 and interleukin 10 in progressing and regressing areas of primary melanoma. AB - The coexistence of tumor specific immunity with a progressing tumor remains a major paradox of tumor immunology. This enigma is most evident in partially regressing melanomas, where efficient eradication of tumor cells is closely linked to uncontrolled tumor growth. Mechanisms involved in this differential susceptibility of tumor cells to the host immune response may include altered production of immunosuppressive cytokines, i.e., transforming growth factor (TGF) beta or interleukin (IL) 10. Since only limited amounts of tissue samples are available from primary tumors, a semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was established which allowed to estimate the amount of cytokine mRNA expressed in a small number of melanoma cells segregated by indirect immunomagnetic isolation. Thereby, we determined the expression of TGF-beta 1 and IL-10 mRNA in melanoma cells obtained from regressing and progressing areas of 9 primary tumors. TGF-beta 1 mRNA could be detected in all undiluted samples from progressing areas and in 7 samples from regression zones. Titration of the sample revealed that in 6 cases TGF-beta 1 mRNA could be detected at a significant higher titer in progressing than in regressing areas. IL-10 mRNA was present in 8 samples obtained from progressing and in 7 samples from regressing tumor areas. In 6 tumors IL-10 mRNA was detectable at a higher titer in the progression zones. Specificity of the PCR amplification was confirmed with a series of restriction enzyme digestions of the resulting PCR product. Based on these findings the hypothesis that immunosuppressive cytokines, such as TGF-beta 1 or IL-10, represent important factors for the melanoma cells to escape immune surveillance is supported. PMID- 10464713 TI - Establishment of a highly metastatic human ovarian cancer cell line (HO-8910PM) and its characterization. AB - To establish a highly metastatic human ovarian cancer cell line and to study its characteristics tissue from the tumor mass of a nude mouse of 7th subtransplantation of the highly metastasizing human ovarian cancer model (NSMO) was cultured in vitro and the cell line HO-8910PM was established. The cell line grew well through 87 passages and the mitotic index, chromosome analysis, morphology under electron microscope and some oncoprotein expression were studied. The cell doubling time was 34.5 h and the mitotic index was 44.3%. The cells were all of epithelial type and most of them were of polygonal shape. Electron microscopic examination showed malignant nuclei with enlarged nucleoli and abundant microvilli. The plating efficiency in soft agar was 31.2%. The cell agglutination appeared in 4 ug/ml PHA. Chromosomal analysis revealed a mode of 54 per cell. The DNA index was 1.57 measured by FCM. Both of them showed hyperdiploid. Positive ER and PR granules were found in the cells. After hetero transplantation of the cells into three nude mice all of the latter showed tumor growth with metastasis in lungs or lymph nodes. Eight of the nine kinds of oncoprotein detected by immnohistochemical method were found in the cells. The detection for mycoplasma showed negative. After storage in liquid nitrogen cell growth was stable. The cell line HO-8910PM can meet the criteria for the establishment cell lines. This cell line and NSMO model would be very useful in study of the mechanism of cancer metastasis in identifying various cellular factors regulating local and distant metastasis and also in establishing a rational approach for searching after anti-metastatic agents. PMID- 10464714 TI - The estimation of endogenous tumor necrosis factor alpha and cortisol levels in serum in advanced neoplasm. AB - The immune reaction mediators--cytokines may play an important role in central nervous system function and influence the neuroendocrine system. The aim of the study was to assess the hypothesis of a possible correlation between endogenous TNF-alpha and glucocorticoids secretion in advanced neoplasm. Thirty patients (17 males and 13 females) with advanced neoplasm were enrolled in the study. The levels of TNF-alpha were measured with ELISA type test and cortisol levels were estimated by RIA method. The blood samples were collected 6 times during 24-hours for TNF and 6 times a day for cortisol. Twenty healthy volunteers were studied in parallel. Both TNF-alpha and cortisol levels revealed the presence of circadian rhythm in examined patients. The levels of TNF-alpha were very high and achieved the peak value about midnight (acrophase-01.40). Cortisol peak was found typically at 08.08. The fluctuations of examined molecules showed regular patterns. TNF-alpha levels markedly decreased together with the increase of cortisol and rose when cortisol concentration in serum was low. Although the correlation was not confirmed by Spearman correlation rank the shapes displayed by both curves may suggest the presence of neuroendocrine feed-back loop between corticosteroids and TNF-alpha in advanced cancer. PMID- 10464715 TI - GABA level and GAD activity in human and mouse normal and neoplastic mammary gland. AB - Our studies have demonstrated that GABA (gamma-aminobutyric acid) is detectable both in mouse and in human normal mammary gland and in neoplastic alterations. We have also shown that GABA content in tumor was significantly higher than in normal tissue. The statistically significant difference in GAD (glutamine acid decarboxylase) activity between tumor and normal mammary tissue was also detected. The positive correlation between GABA content and GAD activity in tumor cells was observed both in human and in mice materials. The observed increase in GABA level and GAD activity in tumor tissue could reflect an eventual local antitumor immune response, however, a hypoxia of tumor cells could also be considered. The role of GABA, GAD and GABA-ergic receptors in cancerogenesis and in cancer progression is still to be clarified and requires further studies; however, it may indicate that the known agonists of GABA-ergic system (e.g. baclofen) can potentially modulate the tumor growth. PMID- 10464716 TI - Inhibitory effect of intestinal bacteria on spontaneous multiple polyps in the small intestine of gnotobiotic BALB/c mice. AB - The incidence of multiple polyps and number of polyps per mouse were significantly lower in conventionalized (CVz), and chloroform-resistant bacteria (CRB)- or fusiform bacteria (FB)-associated mice than in germfree (GF) BALB/c mice. The concentration of fecal fatty acids was also higher in mice associated with either CRB or FB than in GF mice. The incidence of multiple polyps and number of polyps per mouse were significantly correlated with the concentration of fatty acids in the feces of CVz and CRB-GB mice. This study demonstrated that inhibition of multiple polyps in the small intestine of BALB/c mice was affected by the production of intestinal fatty acids. PMID- 10464717 TI - Inhibitory effects of ursodeoxycholic acid on N-methylnitrosourea-induced colon carcinogenesis and colonic mucosal telomerase activity in F344 rats. AB - Bile acids are known to promote colon carcinogenesis. However, one study showed that ursodeoxychlic acid (UDCA) prevented azoxymethane-induced rat colon tumorigenesis. The aim of the present study with 3 sets of experiments was to explore the inhibitory effect of UDCA supplemented in the diet on colon carcinogenesis induced by the intrarectal administration of N-methylnitrosourea (MNU) in F344 rats. In experiment I, 5 rats per group were fed a diet supplemented with 0% (control), 0.4%, 0.08% or 0.016% UDCA or chenodeoxycholic acid (CDCA) for 5 weeks after receiving 3 intrarectal doses of 4 mg MNU in week 1. The formation of colonic aberrant crypt foci (ACFs, preneoplastic lesions) at week 6 showed a 24% and 23% reduction in the 0.4% and 0.08% UDCA groups, respectively, as compared to the control group, while it increased for the 0.4% and 0.08% CDCA groups, and was unaffected in the 0.016% UDCA and CDCA groups. In experiment II based on the results of experiment I, all rats received an intrarectal dose of 2 mg MNU 3 times a week for 3 weeks, and then were administered with 0%, 0.4% or 0.08% UDCA for 27 weeks. At week 30, the incidence of colon tumors in the UDCA groups was significantly lower than that in the control group: 20/50 (40%) and 9/25 (36%) vs. 17/25 (68%). The number of large sized ACFs with 4 or more ACs showed a 47% and 59% reduction in the normal appearing mucosa in the UDCA groups as compared to the control group, while the number of small-sized ACFs with 1-3 ACs was similar in all groups. The normal appearing mucosa showed a noticeable level of telomerase activity (semiquantitative PCR-based TRAP assay) in the control group, and significantly reduced levels in the UDCA groups compared to the control group: 19.8 and 32.7 vs. 71.0 TPG unit in mean value. The colon tumors showed a high level of enzyme activity in both the control and UDCA groups. In experiment III, 6 rats per group were fed a diet supplemented with 0%, or 0.4% UDCA or CDCA for 5 weeks after receiving 3 intrarectal doses of 4 mg MNU in week 1. Two control groups did not receive any treatment with MNU and bile acids. The MNU-treated groups showed significantly elevated levels of colonic mucosal telomerase activity at week 6 as compared to the control group (6.5 TPG unit in mean value). It was noted that both UDCA and CDCA administration reduced the enzyme activity as compared to the group with MNU treatment alone: 24.7 and 25.2 vs. 40.1 TPG unit in mean value. Thus, the present study suggested that orally administered UDCA inhibited the growth of ACFs and the development of carcinomas in the colon of rats treated with MNU. Also, UDCA may suppress MNU-induced telomerase activation in normal appearing but ACF-containing colon mucosa, and its mechanism appears to be different from that responsible for the anti-tumor promoting action of UDCA. PMID- 10464718 TI - The technology explosion in musculoskeletal care--tread slowly. PMID- 10464719 TI - The effect of regular callus removal on foot pressures. AB - The optimum frequency of callus removal is unknown. This study shows that in-shoe dynamic plantar pressure measurement can quantify the effect of callus removal in relation with the frequency of podiatric treatment in diabetic ulcer patients. Three groups of diabetic neuropathic patients were studied. Group 0 consisted of six patients who presented for the first time ever for callus removal and who did not have a history of ulceration. The other two groups had a history of ulceration and required regular podiatry treatment: every 6-8 weeks for 10 patients in group A, and every 3-4 weeks for eight patients in group B. Peak plantar pressures were measured using the F-SCAN system, before and after removal of callus, which was weighed to further quantify the effect of podiatry. When callus was removed, peak pressures fell by 32.1 +/- 8.4% (p = .014) in group 0, by 30.9 +/- 4.5% (p < .005) in group A, and by 24.8 +/- 4.0% (p = .005) in group B. The mean weight of callus removed was not significantly different in the three groups. This suggests that the weight of callus is unlikely to be a reliable guide to assess the time intervals between podiatry appointments. However, the measurement of plantar pressures using a dynamic in-shoe plantar pressure device has proven to be a functional, objective, and quantifiable indicator in the assessment of podiatric treatment. PMID- 10464720 TI - Pneumatic ankle tourniquets: physiological factors related to minimal arterial occlusion pressure. AB - This study investigates several physiological measurements for their correlation to the minimal arterial occlusion pressure using the pneumatic ankle tourniquet. Blood pressure (brachial), height, weight, body fat percentage, ankle circumference, and leg circumference measurements were collected from 50 normotensive healthy subjects. Ankle brachial index (ABI) was also compared. A pneumatic ankle tourniquet was applied to the right ankle and inflated until Doppler insonation confirmed that arterial occlusion had occurred. Mean arterial occlusion pressure was 161.7 mm Hg (N = 50, Nmales = 33, Nfemales = 17), and statistically less than 170 mm Hg (p < .05). The authors present a data model to predict the lowest necessary ankle tourniquet pressure for this study group. Forward stepwise linear regression (critical significance = .01) yields two variables, ankle brachial index and systolic blood pressure, which are used to predict occlusion pressure in this study group. PMID- 10464721 TI - Is glass visible on plain radiographs? A cadaver study. AB - Eighteen randomly selected pieces of nonleaded glass from a collection of 30 pieces from broken bottles of four known color types found on the streets of Houston were sorted into four sets with one control. The purpose of this study was to determine if regular glass is visible on plain radiographs and whether color, location, or volume of these fragments had any effect on the detection of nonleaded glass. These sets were then placed into a fresh-frozen cadaver foot and ankle with a history of insulin-dependent diabetes and peripheral vascular disease. This foot was radiographed utilizing four standard foot projections. Five examiners read these five sets of radiographs twice and recorded the number seen. Overall sensitivity for all of the examiners was 90% with an overall false positive rate of 10%. Intraobserver and interobserver Pearson's correlation coefficients showed that there was reliability between the first and second readings and between observers. The authors concluded that regular nonleaded glass is radiographically visible and that factors such as color and location of the glass have no effect on its detection, while a volume of less than 15 mm3 may have an effect on the detection of glass. PMID- 10464722 TI - Traumatically induced inclusion cyst secondary to shoe impingement: report of three cases. AB - Three cases of traumatically induced inclusion cyst affecting the dorsal aspect of the first metatarsophalangeal joint are presented. Though each case was unique, the clinical presentation was similar with patients relating discrete callus formation with a central sinus overlying an enlarging mass. In all cases, the lesion site involved the dorsal first metatarsophalangeal joint and specifically corresponded to the throatline of "pump" shoes that these patients commonly wore. Predisposing factors in these cases appeared to be forefoot equinus deformity with extensus contractor of the extensor hallucis longus tendon and use of pump shoes with a constricting throatline. The authors describe a surgical approach for treatment of this condition, which addresses underlying etiologies of inclusion cyst formation. Pathology was consistent with traumatically induced inclusion cyst. Though inclusion cysts are common in the foot, rarely is the dorsum of the foot involved, and this article, to the authors' knowledge, is the first to be reported in the podiatric literature. PMID- 10464723 TI - Primary lymphoma of the calcaneus with recurrence in the distal tibia: a case report. AB - Primary lymphoma of bone in general is a very rare condition which represents approximately 3% of all primary malignant bone tumors. Of these, less than 1% originate in the foot. The following case presentation demonstrates a patient who presented with primary lymphoma of the calcaneus, diagnosed by open bone biopsy and treated with radiation therapy. The patient remained disease free for almost 3 years, at which time recurrence to the contralateral distal tibia was diagnosed again through open biopsy. PMID- 10464724 TI - Dystonia: a case report and review. AB - Dystonia is a movement disorder characterized by increased muscular tone for which surgical treatment has met with less than ideal results. Surgical treatment in patients with dystonia is often avoided due to the relative success of medical therapy. However, if expectations stay realistic and if the patient with dystonia is continuously evaluated, appropriate surgical measures may be taken when necessary to optimize patient satisfaction. The authors present a case of successful surgical treatment in a patient with a relatively static but crippling lower extremity deformity. A review of dystonia is included with successful surgical treatment of this compound neurologic disease causing lower extremity deformity in a young female. Diagnosis and treatment options including surgical planning and principles in the dystonic patient are highlighted. PMID- 10464725 TI - Onychomycosis: a compendium of facts and a clinical experience. AB - Fifty percent of all nail disturbances result from onychomycosis, with toenails being affected approximately four times as often as those of the fingernails. The overall incidence in the population is not known, but it is reported at about 2% 13% and is presently on the rise. This lengthy paper reviews all facets of onychomycosis. The present-day classification of onychomycosis is presented. Predisposing factors, causative organisms, as well as the differential diagnosis are included. A thorough comparison of the five presently available oral antifungal agents (griseofulvin, ketoconazole, itraconazole, terbinafine, and fluconazole) in an easy-to-read table is presented. A second table presents many studies performed by many researchers in various countries on these same drugs. The author has included her own personal clinical experience using both fluconazole and terbinafine in 22 and 51 patients with pedal onychomycosis, respectively. This is not meant to be a "controlled study," but rather the experience of one podiatrist in an office setting. The newer oral antifungal agents (itraconazole, terbinafine, and fluconazole) are highly recommended even for the most severe cases of onychomycosis. PMID- 10464726 TI - A simple technique for cannulated screw fixation. PMID- 10464727 TI - Talonavicular joint arthrodesis and Evans calcaneal osteotomy for treatment of posterior tibial tendon dysfunction. PMID- 10464728 TI - The case of glaucoma. PMID- 10464729 TI - Optic cup deepening spatially correlated with optic nerve damage in focal normal pressure glaucoma. AB - PURPOSE: To evaluate whether deepening of the optic cup in patients with focal normal-pressure glaucoma is correlated with the location of most marked loss of neuroretinal rim and visual field. METHODS: Using morphometric evaluation of color stereo optic disc photographs of 102 eyes of 65 patients with focal normal pressure glaucoma, the superior half of the optic disc was compared with the inferior half. RESULTS: In eyes in which the optic cup was deepest in the inferior half of the disc, the most pronounced rim loss was located inferiorly significantly more often than superiorly, and the most marked visual field loss was located superiorly significantly more often than inferiorly. In eyes in which the optic cup was deepest in the superior half of the disc, the most pronounced rim loss was located superiorly significantly more often than inferiorly, and the most marked visual field loss was located inferiorly significantly more often than superiorly. Correspondingly, in eyes in which the most marked rim loss was located inferiorly, the deepest optic cup part was located inferiorly significantly more often than superiorly, and vice versa. CONCLUSION: In focal normal-pressure glaucoma, location of the most marked deepening of the optic cup is spatially correlated with the location of most pronounced neuroretinal rim loss and visual field damage. Because high-pressure glaucoma is typically associated with optic cup deepening and vascular optic nerve damage is associated with optic cup flattening, the spatial correlation between focal optic nerve damage and focal cup deepening may suggest the presence of a pathogenetic aspect in both high-pressure glaucoma and focal normal-pressure glaucoma. PMID- 10464730 TI - Interobserver agreement of Heidelberg retina tomograph parameters. AB - PURPOSE: To measure the interobserver agreement of Heidelberg Retina Tomograph (HRT; Heidelberg Engineering, Heidelberg, Germany) parameters as a result of different observers' contour line placement. METHODS: The optic nerve heads of 50 patients with glaucoma were imaged with the HRT. Five observers traced each disc margin with a contour line. Each observer was masked to the contour line tracings of the other observers, and there was no formal discussion as to where to place the contour line. The following stereometric parameters were calculated for each image for each observer: disc area, mean height of contour, cup shape, rim volume using the standard reference plane from software version 1.11, rim volume using a reference plane of 320 microns below the retinal plane, and volume above curved surface. Agreement between the five observers was tested for each parameter using intraclass correlation coefficients (ICCs). RESULTS: Interobserver agreement between the five observers was substantial for both rim volumes (ICC = 0.73) and for disc area (ICC = 0.67). Agreement was almost perfect for mean height of contour (ICC = 0.94), cup shape (ICC = 0.92), and volume above curved surface (ICC = 0.83). CONCLUSION: The interobserver agreement for the HRT parameters was substantial to almost perfect, indicating that the HRT results as defined by the five observers were interchangeable. PMID- 10464731 TI - Increasing sampling density improves reproducibility of optical coherence tomography measurements. AB - PURPOSE: Published series of peripapillary retinal nerve fiber layer (RNFL) measurements using optical coherence tomography (OCT) have sampled 100 evenly distributed points on a 360 degrees peripapillary circular scan. The goal of this study was to determine whether a four-fold increase in sampling density improves the reproducibility of OCT measurements. METHODS: Complete ophthalmic examinations, achromatic automated perimetry, and OCT imaging were performed in all patients. The OCT scanning consisted of three superior and inferior quadrantic scans (100 sampling points/quadrant) and three circular scans (25 points/quadrant). The RNFL thickness measurements and coefficient of variation (CV) were calculated for the superior and inferior quadrants for each sampling density technique. RESULTS: The study included 22 eyes of 22 patients (3 control subjects; 2 patients with ocular hypertension; and 17 patients with glaucoma). Quadrants with associated glaucomatous visual field loss on automated achromatic perimetry had thinner RNFLs than quadrants without functional defects for both the 25- and 100-points/quadrant scans. For quadrants associated with normal visual hemifields (n = 22), there was no difference between the 25- and 100 points/quadrant scans in mean RNFL thickness and CV. Among quadrants with visual field defects (n = 22), RNFL thickness measurements were thinner in the 25 points/quadrant scans than in the 100-points/quadrant scans. The CV for the 25 points/quadrant scans (25.9%) was significantly higher than that for the 100 points/quadrant scans (11.9%). CONCLUSION: Increasing the sampling density of OCT scans provides less variable representation of RNFL thickness. The optimal sampling density to achieve maximal reliability of OCT scans remains to be determined. PMID- 10464733 TI - Surgical management of the symptomatic overhanging filtering bleb. AB - PURPOSE: Large filtering blebs that evolve after trabeculectomies can be bothersome to the patient, especially when overhanging the cornea. Partial bleb excision is warranted to relieve the patient from discomfort or even visual impairment. METHODS: Surgical partial excision of the overhanging corneal part of the bleb was performed in four patients who had undergone earlier trabeculectomy without application of antimetabolites. Duration of follow-up after excision ranged from 9 months to 4 years. RESULTS: Successful reduction of the excessive bleb and continued satisfactory control of intraocular pressure (IOP) were achieved in all four cases. Partial excision of the corneal part of the bleb did not lead to bleb leakage in any of the cases. CONCLUSION: Surgical blunt dissection of the overhanging the morphologic features of the bleb and ensuring continued control of IOP and relief of symptoms. Alternative methods, such as autologous blood injection, cryoapplication, application of trichloracetic acid, or application of Nd:YAG laser, are noninvasive but do not allow precise rearrangement of bleb architecture. PMID- 10464732 TI - A comparative study of the chemical stability of various mitomycin C solutions used in glaucoma filtering surgery. AB - OBJECTIVE: To determine the chemical stability of various mitomycin C (MMC) solutions used in glaucoma filtering surgery. METHODS: A survey of the MMC solutions currently in use in 21 hospitals (11 in Canada, 10 in the United States) was conducted. A comparative study of the chemical stability of five different representative solutions was performed. The effects of buffer and storage variables on the chemical breakdown of MMC in the solutions were studied by means of high-performance liquid chromatography (HPLC). RESULTS: The survey revealed 33 different variations (including recipes and storage conditions) in the preparation of MMC solutions. Although the majority of the hospitals (15 of 21; 72%) were preparing stable solutions, six of the hospitals (28%) were preparing potentially unstable solutions. The stability of the solutions varied in a nonuniform manner when stored at different temperatures in different buffers. CONCLUSION: The lack of standardization and quality control of MMC solutions used in filtering surgery allows for the possibility of hospitals preparing unstable solutions. PMID- 10464734 TI - The inheritance of the pigment dispersion syndrome in blacks. AB - PURPOSE: Evidence has indicated that pigment dispersion syndrome (PDS) is inherited as an autosomal dominant disorder in white patients, often with a high degree of penetrance. Because heredity patterns in blacks are unknown, an investigation was carried out to study inheritance of PDS in this population. METHODS: Six unrelated black adults (5 women, 1 man, age range 43-60 years) with PDS were identified from a primary eye care population at an inner city teaching clinic in Chicago, Illinois. Nineteen first-degree relatives (all siblings or children; age range 18-52 years) of these patients subsequently underwent thorough eye examination to look for signs of PDS. RESULTS: Among the relatives, two (12%) showed evidence of the condition (these two patients belonged to different families): one was a 42-year-old daughter of a 60-year-old proband, and the other was the 49-year-old sister of a 47-year-old proband. Both exhibited definite signs of PDS in one eye only. CONCLUSION: Evidence of expression of PDS among family members of black probands with PDS is provided. Incomplete penetrance of PDS among the black pedigrees may be suggested by these data. PMID- 10464735 TI - Interaction of pilocarpine with latanoprost in patients with glaucoma and ocular hypertension. AB - PURPOSE: To study any interaction between pilocarpine and latanoprost when administered together, and to determine the optimal timing of dosage to maximize reduction of intraocular pressure (IOP). METHODS: Nineteen adult patients with either primary open-angle glaucoma or ocular hypertension participated in a single-center, prospective case study with masked observer. After a baseline measurement of IOP during treatment with latanoprost was obtained, initial treatment with pilocarpine three times daily was added without bedtime administration. This was followed by three different dose regimens in which pilocarpine was administered four times daily, altering the bedtime pilocarpine dose to precede the latanoprost dose by 1 hour, or to follow it by 10 minutes or 1 hour. Intraocular pressure was measured at 8:00 AM and 75 minutes after administration of the morning dose of pilocarpine. RESULTS: Comparison of IOP at 8:00 AM with baseline showed no significant change when pilocarpine was taken three times daily, or when pilocarpine was taken four times daily when the bedtime dose preceded administration of latanoprost by 1 hour. There were significant decreases in IOP versus baseline when the bedtime dose of pilocarpine was taken simultaneously with or 1 hour after administration of latanoprost. Application of pilocarpine immediately after the 8:00 AM IOP measurement revealed a significant additional decrease in pressure. There were no significant differences between dosage schedules in the magnitude of the additional reduction in IOP. CONCLUSION: The order and timing of administration of pilocarpine and latanoprost can significantly alter their ocular hypotensive activity. Pilocarpine is most effective when administered four times daily, and when the bedtime dose is administered 1 hour after administration of latanoprost. PMID- 10464736 TI - Management of late bleb leaks. PMID- 10464737 TI - Ethical considerations related to outcome studies-based clinical practice guidelines. AB - Expanding demand for health services is occurring in the face of diminishing resources. Root causes for this phenomenon include advancing expensive new technology and marked variations in quality of care and patterns of practice. Practice guidelines resulting from valid and appropriate outcome studies have the potential to promote consistency and quality. Perceived drawbacks to such guidelines include over-regulation and loss of autonomy for both physicians and patients. Clinical practice guidelines are discussed in the context of major ethical theories, including deontology, utilitarianism, and contractarianism. The conflicting needs and motivations of physicians, patients, third-party carriers, and society as a whole are discussed relative to these different ethical perspectives. Although contrasting ethical foundations likely will lead to significantly different solutions to the healthcare resource problem, it is only through educated and reasoned discussion that this difficult problem can be tackled. Sound ethical discipline is imperative when considering the related issues of equity, justice, and autonomy in a society with progressively decreasing resources. PMID- 10464739 TI - Bilateral Chandler syndrome. PMID- 10464738 TI - A randomized study of dorzolamide in the prevention of elevated intraocular pressure after anterior segment laser surgery. Dorzolamide Laser Study Group. AB - PURPOSE: To evaluate the efficacy of 2% dorzolamide hydrochloride opthalmic solution in preventing spikes in intraocular pressure (IOP) after anterior segment laser surgery. METHODS: This 24-hour, placebo-controlled, randomized, double-masked, multicenter evaluation was conducted to determine the efficacy of dorzolamide hydrochloride 2% in controlling IOP after neodimium:yttrium-aluminum garnet (Nd:YAG) laser capsulotomy, argon laser trabeculoplasty, or laser iridotomy. The 122 patients enrolled were assigned in randomized fashion to receive dorzolamide or placebo 1 hour before and immediately after the procedure; IOP was measured 1, 2, 3, 4, and 24 hours after the procedure. RESULTS: Of 61 patients receiving dorzolamide, only one (1.7%) had a spike in IOP of 10 mmHg or more, compared with 9 (14.8%) of the 61 patients receiving placebo. Mean IOP among patients receiving dorzolamide was significantly reduced both from baseline and compared with that among patients receiving placebo from 1 to 4 hours after administration. Only 5 (8%) of the 61 patients receiving dorzolamide experienced at least one adverse event, compared with 15 (25%) of the 61 patients receiving placebo. CONCLUSION: Dorzolamide was effective in preventing spikes in IOP after anterior segment laser surgery. Dorzolamide was generally well tolerated after short-term use. PMID- 10464740 TI - [Reimplantation lesion in the postoperative period of lung transplantation. Incidence, predictive factors, prognosis and outcome. The Lung Transplantation Group of Vall d'Hebron University General Hospital]. AB - BACKGROUND: Reimplantation disease (RD) is a postoperative complication in lung transplantation. It is defined as hypoxemia (PaO2/FiO2 ratio < 150 mmHg), radiologic infiltrates and decrease of lung compliance. The aim of the study was to analyze the incidence, predictive factors, prognosis and outcome of the patients with RD. PATIENTS AND METHOD: 49 patients submitted to lung transplantation (June 1991-December 1996) were admitted in our intensive care unit (ICU). Donor and recipient conditions, surgical parameters and outcome in ICU were analyzed. Mann-Whitney, Kruskall-Wallis, Fisher, Pearson and ANOVA Friedman tests were used for statistical analysis according to the different variables. RESULTS: 49% of the patients (29/49) developed RD, which was influenced neither by lung disease, nor by the kind of transplantation or by ischemia time. All patients with a long surgical time developed RD, versus only 41% in those where surgery was undertaken in a shorter period of time, OR: 2.8 (1.5-5.7; p = 0.0016). The patients with RD improved showing a PaO2/FiO2 ratio of 176 and 235 mmHg at 24 and 48 h respectively (ANOVA, p < 0.00001). The patients with RD needed 14 days of mechanical ventilation versus 7 days in those without RD (p = 0.013). There were no statistically significant differences in stay and mortality in ICU. CONCLUSIONS: RD is a common complication in the postoperative phase of lung transplantation. It is present in almost all the patients with long surgical time. Almost all of them improve, with the same survival but a longer period of mechanical ventilation. PMID- 10464741 TI - [Community-acquired pneumonia: impact of the use of a therapeutic strategy based on probability of short-term mortality]. AB - BACKGROUND: To assess the impact of the use of a therapeutic strategy based on classifying patients with community-acquired pneumonia (CAP) according to the probability of short-term mortality. PATIENTS AND METHODS: During one year, all patients admitted to the Emergency Department with diagnosis of CAP were included. Clinicians were invited to treat patients according to a recently published protocol that stratifies patients into five categories (from low to high-risk mortality): patients assigned to class 1 were managed at home; patients included in classes 2 and 3 were assigned to a short-time period at emergency department before managed at home; and patients assigned to classes 4 and 5 were hospitalized. RESULTS: The final population analyzed included 101 patients. The rate of acceptability among clinicians was 96.7%. Patients were classified by the following terms: risk-class 1: 17 (16.8%); risk-classes 2 and 3: 40 (39.7%); risk classes 4 and 5: 44 (43.6%). During follow-up, of the 57 non-hospitalized patients, 3 (5.2%) were subsequently admitted to hospital and 7 (12.2%) patients initially assigned to a short-time period at emergency department were hospitalized, and 1 (1.7%) of them died. By this program, the reduction of the hospitalization rate was 23.8%. CONCLUSION: A strategy of management of CAP based on a prognostic classification has a good safety and acceptability among clinicians, and reduces the rate of hospitalizations. PMID- 10464743 TI - [Lung transplantation 1999]. PMID- 10464742 TI - [Withdrawal of Pneumocystis carinii pneumonia prophylaxis in patients receiving efficacious combined antiretroviral treatment. Study of 85 cases]. AB - OBJECTIVES: To analyse the incidence of Pneumocystis carinii pneumonia after the withdrawal of prophylaxis, in patients with AIDS who were receiving HAART (highly active antiretroviral treatment). PATIENTS AND METHODS: Prospective opened study of 85 patients with AIDS (CD4 lymphocytes < 200 x 10(6)/l) and whose CD4 counts had increased over 200 x 10(6)/l after HAART, 79 were under primary prophylaxis and six secondary. RESULTS: Mean CD4 lymphocytes count at the time of withdrawal was 343 x 10(6)/l. Mean time of follow-up after withdrawal was 358 days (range: 93-1,487; median: 302). None of the patients have had Pneumocystis carinii or toxoplasmosis after withdrawal of prophylaxis. CONCLUSIONS: Pneumocystis carinii prophylaxis in AIDS patients might be safety withdrawn after effective HAART. PMID- 10464744 TI - [Interactions of antiretroviral drugs]. PMID- 10464745 TI - [Pharmacodynamic assessment of antibiotic sensitivity of Streptococcus pneumoniae]. PMID- 10464746 TI - [Pulmonary infiltrates in a 46-year-old man recipient of an allogeneic transplant of bone marrow (clinico-pathologic conference)]. PMID- 10464747 TI - [Efficacy and safety of low molecular weight heparin in the treatment of pulmonary thromboembolism]. PMID- 10464748 TI - [Subcutaneous panniculitic T-cell lymphoma]. PMID- 10464749 TI - [Homonymous quadrantanopia as a presentation form of multiple sclerosis]. PMID- 10464750 TI - [Sister Mary Joseph's nodule]. PMID- 10464751 TI - [Internet guide on instructions to authors of more than 2000 biomedical journals: Raymon H. Mulford Library]. PMID- 10464752 TI - [An algorithm for construction of a mathematical model of coronary circulation disorders]. AB - The paper shows how to improve the quality of diagnosing coronary heart disease (CHD). It proposes a mathematical model of coronary circulatory disorders, which is based on the definition of CHD as a cardiac abnormality caused by the imbalance of coronary blood flow and myocardial oxygen consumption, outlines an algorithm for construction of this model. With this model, a computer system has been designed for examining a patients with CHD, which permits the construction of a virtual model of coronary circulatory disorders. PMID- 10464753 TI - [Mathematical model for prognostication of work capacity recovery in patients with myocardial infarct]. AB - The impact of clinical and instrumental findings on prognosis was evaluated in patients with myocardial infarction and a mathematical model was proposed to predict their recovered working ability. The developed predictive index for prediction of recovered working ability is of high informative value and accuracy. It may be practically used by a cardiologist to choose a protocol of medical and rehabilitative measures in myocardial infarction. PMID- 10464754 TI - [Goals of medical monitoring]. AB - The paper deals with the problems of medical monitoring, namely how to solve those in the detection and classification of human abnormalities. The medical monitor is a complex hierarchic system and the authors state problems and tasks for each level of hierarchy and the objectives of studies in medical diagnosis and prediction while designing the hierarchic network structure of a medical computer monitor. A parallel is drawn between the image recognition theory and medical diagnosis. PMID- 10464755 TI - [Cardiac interval measurement attachment to a personal computer]. AB - A cardiac intervalometric deck to a personal computer is described, which can single out respiratory arrhythmia in the pattern of cardiac rhythm. The parameters of respiratory arrhythmia supplement variation pulsometry to evaluate the autonomic nervous system. PMID- 10464756 TI - [A decisive rule in classifying diseases of the visual system]. AB - The paper deals with a new method for recognizing images, which is based on the use of deciding rules to classify a difficult area of diagnosis of eye abnormalities. PMID- 10464757 TI - [Laboratory diagnosis of changes in regulation of the immune system]. AB - To evaluate the body's immunity, to diagnose immunodeficiencies is a pressing problem. The paper discusses whether a complex approach can be used to evaluate immunity objectively. The data on humoral and cellular immunity in patients with varying responses to the changes occurring in the body are analyzed. The changes in cellular and humoral immunity were studied by enzyme immunoassay of hormones and antibodies in combination with immunomorphological assay of lymphocytic subpopulations. PMID- 10464758 TI - [Current trends and problems in the measurement of limb loading parameters during transosseous osteosynthesis using external fixation devices]. AB - The problem in question is whether it is reasonable to equip a conventional limb external fixation apparatus with means for the diagnosis of loading parameters: the forces and displacement of the opposite supporting elements of the apparatus. It is assumed that during distraction osteogenesis it is expedient to apply load to the limb by the load rate only at the initial stage of callus regeneration (the plastic or slightly elastic mechanical state) while the most effective way to do so is to act force at later stages. A number of functions derived from the modified parameters which might aid a physician to choose the optimum limb loading parameters at different stages of treatment were considered by using as example a hypothetical ring-type apparatus equipped with means for continuous measurement of current forces applied to the apparatus and current displacements of opposite ring blocks. Specifications for the measuring system of loading parameters were defined by using an Ilizarov apparatus as an example and why possible rough measurement errors may occur is revealed. The paper shows it necessary to develop methods for calculation of parasitic deformations, i.e. those of the loaded elements of the apparatus for the evaluation of the reciprocal displacement of bone fragments. PMID- 10464759 TI - [Current possibilities of routine fluorography]. AB - New Russian 12 phi 9 fluorographic apparatuses with a K phi -400 fluorographic camera are shown to have some advantages over a 12 phi 7 fluorograph. The fluorographs ensure a much better image quality in a smaller input operating dose. The use of the new generation fluorographs provides for a patient a radiation load approximate that of direct film X-ray study. PMID- 10464760 TI - [Device-aided determination of an effective dose in x-ray studies]. AB - The paper proposes how to determine an effective radiation dose for patients undergone X-ray examinations, which includes the estimation of the size of a field while measuring the superficial dose and the assessment of an experiment protocol by taking into account the X-ray receiver used to measure exposure. PMID- 10464761 TI - [An instrument for intraosseous punctures]. AB - The paper outlines an intraosseous paracentesic tool, a needle-screw device that combines the properties of a drill and a hollow tube with external thread. The techniques of bone paracentesis and appropriate procedures are described. The results of examining osseous injuries while using various tools to form perforating tools have shown that the needle-screw device has advantages over others. PMID- 10464762 TI - [Synchronous automated analysis of a spirogram and cardiac intervalogram]. AB - The paper deals with the functional diagnosis based on the synchronous analysis of a spirogram and a cardiac intervalogram (CIG) by evaluating respiratory sinus arrhythmia (RSA). A method based on the calculation of R-R interval differences in separate breathing cycles is used for automated reckoning of RSA. A band-pass filter is employed to separate out a respiratory constituent of CIG. The programmed algorithmic use of the method is based on a linear selective transformation that is a general procedure for morphological analysis of cardiac signals in real time. PMID- 10464763 TI - [Computerized technologies in public health administration]. AB - The paper deals with the problems in making the computer monitoring of qualitative, quantitative, structural, and functional indices at all levels of the public health system in the Sverdlovsk Region. The sources of information were defined at all levels of an informational field and informational channels were set up. The monitoring systems are reinforced by the currently available systems and the systems under design. The programme complexes realize the methods of multifactorial analysis, taxonomy, econometry, identification of large systems and they are adjusted to the existing information collection system based on the corporative control network of public health. PMID- 10464764 TI - [History of creation of the doctrine, equipment and methods of formation of biological feedback]. AB - The theoretical and experimental priorities of Russian scientists A. V. Zaporozhets and M. I. Lisina in creating the doctrine of biological feedback (BFB) in 1955 are justified. The priority of American scientists (N. Miller, 1969) in the discovery of the fact that BFB can form in animals is recognized. USA scientists were also be the first to develop and provide a base for manufacturing commercial devices for shaping BFB that have gained wide practical recognition in medicine, sports, and psychophysiology. PMID- 10464765 TI - [Uroflowmeter, a device for measuring urine volume and urination rate]. AB - The paper deals with the design and construction of a float-type uroflowmeter that is characterized by higher reliability, precision, validity of measurements. The characteristics of the device have been improved by excluding a sliding contact which operates in the corrosive medium, by applying a primary gauge whose output signal is proportional to the instantaneous value of urinary debit. PMID- 10464766 TI - Purification and characterization of two recombinant human granulocyte colony stimulating factor glycoforms. Pharmacokinetic and activity studies of single dose administration in mice. AB - Two recombinant human granulocyte colony-stimulating factor (rhG-CSF) isoforms were isolated from the medium conditioned by an engineered Chinese hamster ovary (CHO) cell line. The two rhG-CSFs were characterized and were found to differ in the carbohydrate structure attached to Thr-133. The glycoform, referred to as Peak 1, contains the O-linked glycan Neu5Ac(alpha 2-3)Gal(beta 1-3)GalNAc; the Peak 2 glycoform contains the O-linked glycan Neu5Ac(alpha 2-3)Gal(beta 1 3)[Neu5Ac(alpha 2-6)]GalNAc. The two glycoforms displayed a similar biological activity in cultures of a mouse 32D C13 cell line and human bone-marrow myelo monocytic progenitor cells (CFU-GM). In the latter test both glycoforms displayed a higher activity than nonglycosylated rMet-hG-CSF from Escherichia coli. The pharmacokinetic profile and activity of the two rhG-CSF glycoforms and of a mixture of them (Pool) were investigated in mice treated with a single injection of rhG-CSF at the doses of 125 micrograms and 250 micrograms/kg, given via the intravenous (i.v.) and the subcutaneous (s.c.) route, respectively. The plasma concentration profiles obtained were similar for all three substances and did not show any relevant differences in absorption or elimination. The pharmacokinetic parameters indicate that the three substances have similar area under the curve (AUCs), volumes of distribution, and terminal half-life. Furthermore, our data indicate a high bioavailability of the two different glycoforms of rhG-CSF when given to mice via the s.c. route either singularly or as a mixture. Detectable levels of rhG-CSF persisted for more than 8 h in the i.v. and more than 24 h in the s.c. route of administration. All three substances induced early neutrophilia in mice. All rhG-CSF-treated mice developed a two-four-fold rise in neutrophil counts as early as 4 h after the intravenous and 2 h after the subcutaneous injection. Relatively high levels of neutrophils were maintained for at least 8 and 24 h after i.v. and s.c. administration, respectively. PMID- 10464767 TI - Characterization of transposon-generated protease mutant of Xanthomonas campestris pathovar glycine 8ra. Enzyme activity, cloning, and mapping of flanking DNA. AB - Protease negative mutant of Xanthomonas campestris pathovar glycine 8ra (prt mutant) was constructed by mutagenesis employing artificial transposon Omegon-Km. Transposon delivery was conducted through diparental conjugation using X. campestris pathovar glycine 8ra as recipient and Escherichia coli S17-1 carrying pJFF 3500 plasmid as the donor. The frequency of transconjugation was found 1.9 x 10(-7) per recipient. Enzyme analysis indicated the presence of mutant with lower protease activity than that of the wild-type. Genetic analysis employing pulsed field gel electrophoresis (PFGE) of the genomic DNA digested with AseI or SpeI restriction endonuclease could significantly differentiate X. campestris pathovar glycine 8ra prt from the wild-type parent. The 9.85 kb pLR omega 6 plasmid was constructed from the genomic DNA of the prt mutant after being digested with KpnI restriction endonuclease and ligated with T4 DNA ligase. PMID- 10464768 TI - Rabies vaccine. Developments employing molecular biology methods. AB - Rabies vaccines produced by means of molecular biology are described. Recombinant vaccines employing either viruses as vectors (vaccinia, adenovirus, poxvirus, baculovirus, plant viruses) or a plasmid vector carrying the rabies virus glycoprotein gene are discussed. Synthetic peptide technology directed to rabies vaccine production is also presented. PMID- 10464769 TI - A role for leptin in hemopoieses? AB - The recent discovery of leptin as a major controller of appetite has led to a detailed analysis of its specific actions in this process as well as any potential role in the etiology of obesity. It has also emerged that leptin has a wider spectrum of biological activities and has been strongly implicated in fertility and reproduction. The structural similarity between leptin and its receptor and cytokine-receptor systems that control hemopoiesis has also prompted investigation of the potential for this hormone to influence blood cell formation. Recent studies have shown that the leptin receptor is expressed on a diverse range of hemopoietic cells. Leptin itself appears to enhance proliferation of hemopoietic cells in vitro, particularly in combination with other cytokines and may augment some mature hemopoietic cell functions. Although only relatively minor hemopoietic deficiencies have been reported in mice lacking leptin or its receptor, these emerging studies suggest that further analysis of leptin actions in hemopoiesis may be warranted. PMID- 10464771 TI - New directions in liposome gene delivery. AB - The history of liposomes, progress in liposome gene delivery, and future directions are discussed. Specific characteristics of liposomes and DNA:liposome complexes have been identified that are essential for optimal delivery and gene expression. Of particular interest are the requirements for increased delivery and high levels of gene expression in vivo. At present, significant efforts are focused towards achieving specific delivery and gene expression in target organs and tissues. PMID- 10464770 TI - Genomic imprinting and chromatin insulation in Beckwith-Wiedemann syndrome. AB - Genes are recognized as undergoing genomic imprinting when they are capable of being expressed only from the paternal or only from the maternal chromosome. The process can occur coordinately within large physical domains in mammalian chromosomes. One interesting facet of the study of genomic imprinting is that it offers insight into the regulation of large chromosomal regions. Understanding this regulation involves elucidating the cis-acting regulators of gene expression and defining the elements that maintain chromatin insulation, both required for understanding more practically applicable areas of biological research, such as efficient transgene production. This review is focused on the regulation of the imprinted domain of human chromosome 11p15.5, responsible for Beckwith-Wiedemann syndrome (BWS). Recent findings indicate that the maintenance of imprinting within this domain is critically dependent on the stable maintenance of chromatin insulation. PMID- 10464773 TI - Recovering cDNA bands from differential display RT-PCR gels using a transparency film mask. AB - We describe here the use of a transparency film as a mask for recovering bands from differential display reverse transcription-polymerase chain reaction (DDRT PCR) gels. This method represents a simple and rapid way to isolate the differentially expressed bands from dried and nondried polyacrylamide, radioactive and nonradioactive, denaturing and native DDRT gels. A transparency film is overlaid on the DDRT autoradiogram, and the marks and bands of interest are drawn using a permanent marker. The reproduced band marks are cut out of the transparency sheet with a scalpel to facilitate the recovery of the desired bands. The transparency film mask is then overlaid on the top of the gel and the bands are recovered from the gel. The use of the transparency film mask avoids damage to the autoradiogram and is also extremely useful in DDRT-PCR experiments involving different RNA samples that produce band patterns of different intensities that require many X-ray exposures for different periods of time. PMID- 10464774 TI - Rapid preparation of denatured double-stranded DNA templates for sequencing. AB - This article proposes protocol for rapid preparation (ds) DNA templates for sequencing based on double-stranded DNA denaturation and its recovery by extraction with Wizard DNA purification resin (Promega). This method is an alternative to commonly used procedure employing denatured-DNA recovery by ethanol precipitation. PMID- 10464772 TI - The use of soybean agglutinin (SBA) for bone marrow (BM) purging and hematopoietic progenitor cell enrichment in clinical bone-marrow transplantation. AB - Soybean agglutin (SBA) is a plant lectin that has been used to fractionate bone marrow cells. It binds bone marrow mononuclear cells, including mature myeloid, erythroid, and lymphoid cells, but has very low binding affinity and no toxic effect to the human hematopoietic cells. In this article we describe the possibilities of enriching bone-marrow-derived CD34+ hematopoietic progenitor cells using SBA. As the method is simple and elegant. SBA is of vast importance to the field of clinical bone marrow transplantation. PMID- 10464775 TI - [Is there solid evidence for reorganization of the neural network associated with development of behavioral sensitization to psychostimulants?]. AB - Behavioral sensitization or reverse tolerance induced by repeated treatment of psychostimulants, such as amphetamine and cocaine, has been widely used as an animal model for schizophrenia. This phenomenon may result from almost eternally lasting plastic functional adaptation in the brain. Such long-lasting plasticity must accompany anatomical and morphological neural changes. Histological studies revealed that chronic treatment of psychostimulants induced thickness, elongation and an increased branch number of neurites and dendrites in cortices and accumbens, and an increased spine density. These changes may seem a positive adaptation for neural transmission, however, similar histological changes were seen after toxicity of methamphetamine as deafferation compensation. Therefore, further studies are needed to establish the significance of these morphological changes induced by chronic psychostimulants. On the other hand, neuromolecular studies showed increased phosphorylation of neuromodulin, increased expression of tissue-type plasminogen activator, synaptotagmin IV and arc, which are assumed to play roles in neural outgrowth and synaptogenesis. Although all of these histological and neuromolecular findings may suggest reorganization of the neural network should grow along with behavioral sensitization, more solid evidence is indispensable to confirm it. PMID- 10464776 TI - [Early phase II study of quetiapine fumarate on schizophrenia]. AB - The efficacy and safety of quetiapine fumurate in the treatment of patients with schizophrenia were evaluated in an 8-week, multicenter, open-label study. The results of this study which included a total of 54 patients showed good efficacy and safety profile for quetiapine fumarate as seen by the improvement rate (moderate or above in the final global improvement rating) of 49.1% and safety rate (no problem in overall safety rating) of 66.0%. The mean BPRS total score decreased significantly from 55.5 +/- 10.9 points at baseline to 45.4 +/- 13.0 points at the completion of administration. The PANSS scores also showed significant improvement on all scales; the mean scores decreased from 20.7 +/- 6.3 points at baseline to 17.7 +/- 6.9 points at withdrawal or completion of administration on the positive scale, from 27.8 +/- 5.8 points to 24.0 +/- 7.3 points on the negative scale, and from 51.4 +/- 10.1 points to 44.7 +/- 12.4 points on the general psychopathology scale. Although the most frequent adverse reactions were somnolence (18.9%), insomnia (17.0%), nervousness (13.2%), dizziness (13.2%), malaise (13.2%), postural hypotension (11.3%), tachycardia (9.4%), and constipation (9.4%), the incidence of extrapyramidal symptoms was low (11.3%). From these results, quetiapine fumarate was suggested to be highly effective and safe for the treatment of schizophrenia. PMID- 10464777 TI - [Saliva level of free-3-methoxy-4-hydroxyphenylglycol in patients with anxiety disorders]. AB - The saliva level of free 3-methoxy-4-hydroxyphenylglycol (MHPG) in 16 outpatients with anxiety disorders was determined by gas chromatography-mass spectrometry. The anxiety level of the patients was scored by the Hamilton Anxiety Scale (HAS) before and after a one-week drug treatment with alprazolam. The saliva level of free-MHPG at the subjects' first visit to the hospital was significantly higher than that of the control subjects and was reduced by the one-week alprazolam treatment. The post-treatment level of MHPG in patients who showed good response to the treatment was significantly less than the pretreatment level, but there was no significant difference between pre- and post-treatment levels of MHPG in poor responders. There was no correlation between the MHPG level and the HAS score at the first hospital visit. The MHPG levels after the treatment correlated with the HAS scores. The reduction of the anxiety level as scored by the HAS correlated with the reduction of the MHPG level. These results indicate that the free saliva MHPG level may be a useful indicator for assessing not only the level of anxiety but also the response to drug treatment for anxiety in these patients. PMID- 10464778 TI - [Behavioral pharmacological characterization of mice lacking the nociceptin receptor]. AB - Nociceptin and nociceptin receptor, which show structural similarities to opioid peptides and opioid receptors, respectively, have been recently found to constitute a novel neuromodulatory system. In the central nervous system, however, the physiological role of modulation via the nociceptin receptor is still unclear. Here, we report the behavioral pharmacological characterization of mice lacking the nociceptin receptor. Nociceptin produced hyperalgesia and hypolocomotion, whereas the nociceptin receptor-knockout mice showed no significant abnormalities in nociceptive thresholds (tail-flick, hot-plate, electric, and acetic acid-induced writhing tests) and locomotion. In the learning and memory tests, nociceptin induced impairment of learning and memory in wild type mice. Nociceptin receptor-knockout mice possessed greater learning ability and had better memory than wild-type mice. These results suggest that the nociceptin system plays a role in regulation of nociception or locomotion and seems to play negative roles in learning and memory. Next, we compared nociceptive responses induced by various opioids between the nociceptin receptor knockout and wild-type mice. As previously reported, morphine (mu-opioid receptor agonist), U-50,488 H (kappa 1-opioid receptor agonist), and naloxone benzoylhydrazone (NalBzoH; kappa 3-opioid receptor agonist) induced antinociceptive effects in wild-type mice. Surprisingly, knockout mice lacked the antinociceptive effect induced by NalBzoH, but not by morphine and U-50,488H. Further, NalBzoH completely inhibited nociceptin-induced hyperalgesia and hypolocomotion in wild-type mice. Experiments on the cultured cells transfected with the nociceptin receptor cDNA showed that NalBzoH competed in [3H]-nociceptin binding and attenuated the nociceptin-induced inhibition of cyclic AMP accumulation induced by forskolin. These results clearly suggest that NalBzoH acts as a potent antagonist for the nociceptin receptor. Our studies suggest that the nociceptive system and/or learning and memory processes could be modulated by ligands to the nociceptin receptor, and further that the antagonists are worth testing for the alleviation of pain and memory disorders. PMID- 10464779 TI - [Molecular mechanisms of the pathway formation in the fetal rat cerebral neocortex]. AB - Neural cell adhesion molecules (NCAMs), L1 and TAG-1, which are prominently expressed in the developing nervous system, have been shown to promote axonal growth and bundle formation of central neurons in vitro. In the cerebral neocortex of fetal rats, immunoreactions of L1 and TAG-1 were specifically localized on thalamic afferent axons and cortical efferent axons, respectively. L1-bearing thalamocortical axons preferentially extended in the subplate of the cortical anlage where neurocan, a brain-specific chondroitin sulfate proteoglycan (CSPG), was prominently expressed. In contrast, cortical efferent axons immunoreactive for TAG-1 did not enter the subplate and run in the intermediate zone where neurocan expression was less abundant. In addition, TAG-1-bearing axons extensively invaded regions expressing another type of brain-specific CSPG, phosphacan. In the cell culture system, neurite outgrowth of TAG-1-transfected PC 12 D cells was remarkably inhibited on the neurocan substrate, while the outgrowth on phosphacan substrate was significantly promoted. Although both L1 and TAG-1 have been reported to bind both neurocan and phosphacan in vitro, interactions between NCAMs and CSPGs in vivo indicate more complicated patterns than previously thought. Thus, the present results suggest that various patterns of functional correlation between NCAMs and CSPGs play important roles in the pathway formation of the rat cerebral neocortex. PMID- 10464780 TI - [The relation between metabolism of biopterin and dystonia-parkinsonism]. AB - Tetrahydrobiopterin (BH4) is an essential cofactor for tyrosine hydroxylase. BH4 can be synthesized from GTP through three enzymatic reactions. The rate-limiting step of the BH4 synthesis is catalyzed by GTP cyclohydrolase I (GCH). Recently, we found that GCH is a causative gene for hereditary progressive dystonia/dopa responsive dystonia (HPD/DRD). However, several problems still remain to be solved. The first concern is the presence of asymptomatic carriers in the disease. The difference between symptomatic and asymptomatic carriers is unknown. Second, we cannot find any mutation in the coding region of the GCH gene in about 40% of the patients. What kind of mutation would be present in these patients. The last concern is the molecular mechanism how the enzymatic activity is decreased to less than 20% of normal values. Further studies are required to solve the questions. PMID- 10464781 TI - Effect of a new starch-free bread on metabolic control in NIDDM patients. AB - BACKGROUND AND AIM: The aim of the study was to evaluate the effect on blood glucose levels in non-insulin-dependent diabetics (NIDDM) of reduction of the carbohydrate content through the use of a new, almost starch-free type of bread (SF-bread). We only substituted the bread in the breakfast meal. METHODS AND RESULTS: The study consisted of two parts: 1) a two-day randomized study of the effect of SF-bread on the morning blood glucose levels of NIDDM patients and 2) an open, crossover trial of three months duration where each patient was given SF or ordinary bread. Ten patients participated in the first part and eight in the second part of the study. All patients had well established non insulin-dependent diabetes mellitus. In the first part of the study, the area under the curve describing time-dependent changes in blood glucose level after a standard breakfast was significantly lower in patients on SF-bread (182 +/- 154 Units; mean value +/- SD) than in the controls (630 +/- 258 Units; p < 0.00001). Peak blood glucose concentration was 14.8 +/- 2.3 mM on the control day and 11.6 +/- 1.7 mM on the SF-bread day (p < 0.001). In the second part of the study, the diet including SF-bread reduced fasting blood glucose from 13.3 +/- 3.5 mM to 10.2 +/- 2.0 mM (p < 0.006) and the fraction of HbA1c from 0.090 +/- 0.014 to 0.081 +/- 0.015 (p < 0.02). Similar changes were not seen on the ordinary diet. Serum cholesterol levels were significantly reduced by the SF-bread as compared to the ordinary diet (5.8 +/- 0.6 to 5.5 +/- 0.5 mM versus 5.7 +/- 0.8 to 5.8 +/- 0.7 mM; p < 0.05). CONCLUSIONS: Substitution of ordinary bread with starch-free bread at breakfast causes significant improvements in blood glucose levels in NIDDM patients on both a short and long term basis. Possibly secondary to this, a favorable influence on lipid levels was noted. PMID- 10464782 TI - Effect of virgin olive oil phenolic compounds on in vitro oxidation of human low density lipoproteins. AB - BACKGROUND AND AIM: Substantial evidence suggests that oxidative modifications of low density lipoproteins (LDL) critically contribute to the pathogenesis and progression of human atherosclerosis. Oxidized LDL (oxLDL) are present in atherosclerotic plaques and contain oxysterols that exhibit a variety of adverse biological activities. Antioxidants have also been shown to prevent LDL modification. We have therefore assessed the efficacy of virgin olive oil phenolic compounds in preventing oxidative modifications of human LDL oxidized by UV light. METHODS AND RESULTS: Cholesterol oxides formed during LDL photo oxidation were determined by UV-HPLC in the presence of different concentrations of phenolic compounds and their pure components (tyrosol and oleuropein), and probucol, a widely used synthetic antioxidant. Electrophoretic mobility was also assayed. The results demonstrate that phenolic compounds are much more potent in preventing cholesterol oxide formation and apoproteic moiety modification than their pure components and probucol. CONCLUSIONS: The beneficial effects of a Mediterranean diet may be ascribable not only to the high unsaturated/saturated fatty acid ratio characteristic of olive oil, but also to the unique antioxidant properties of its phenolic compounds. PMID- 10464783 TI - Effect of plasma cholesterol reduction by pravastatin on the functional properties of forearm arteries in hypercholesterolemic patients. AB - BACKGROUND AND AIM: Since functional properties in the vasculature of hypercholesterolemic subjects are impaired, a six-month pravastatin treatment (20 mg/die) was tested in an open design, on the impaired unstimulated forearm arterial compliance (Un-FAC(AUC)) of 14 asymptomatic type IIa familial hypercholesterolemic patients. In order to evaluate whether FAC(AUC) changes might be related to the extent of cholesterol reduction achieved, this evaluation was carried out in five severely hypercholesterolemic patients, undergoing LDL apheresis. METHODS AND RESULTS: Arterial functional properties, i.e. FAC(AUC) responses to glyceryl trinitrate (GTN-FAC(AUC)) and acetylcholine (ACh-FAC(AUC), four patients) and the effects on rest and peak forearm blood flow and vascular resistance were evaluated on the non-dominant arm using plethysmographic methods, that also allow the direct assessment of the non-linear "compliance-blood pressure" curve. Selective LDL-apheresis was performed by using a dextran sulphate column. Pravastatin effectively lowered plasma total (-16%, p = 0.002) and LDL cholesterol levels (-22%, p = 0.006 vs baseline). Rest and peak flow, basal and post ischemic vascular resistance were not affected as well as Un FAC(AUC) and GTN-FAC(AUC). However, in the four hypercholesterolemic patients undergoing ACh infusion, there was an improvement in the ACh-FAC(AUC) of borderline statistical significativity (p = 0.056). LDL-apheresis reduced plasma total and LDL cholesterol levels by 55% and 59%, without affecting blood pressure. In this series of five patients Un-FAC(AUC) increased, the Un-FAC(AUC) rise being inversely related to the absolute reduction of plasma total (r = 0.92, p < 0.05) and LDL cholesterol (r = 0.89, p < 0.05) levels. CONCLUSIONS: In hypercholesterolemic patients a short-term hypocholesterolemic treatment with pravastatin, although able to improve the lipid profile, cannot alter significantly blood flow, vascular resistance, Un-FAC(AUC) and GTN-FAC(AUC). A possible selective improvement in the ACh-receptor-activated signal transduction pathway has been observed and the importance of a drastic reduction of cholesterol concentrations in order to affect the Un-FAC(AUC) is suggested. PMID- 10464784 TI - Role of lipid, apolipoprotein levels and apolipoprotein E genotype in young Italian patients with myocardial infarction. AB - BACKGROUND AND AIM: Studies of young patients with acute myocardial infarction (AMI) have demonstrated that conventional risk factors are usually responsible for their premature atherosclerosis. No account has yet been published of the risk profile of young Italians surviving an AMI. In this study, the conventional risk factors, lipids and apolipoproteins, and apolipoprotein E (APOE) allele distribution were evaluated in 98 consecutive AMI survivors (94 males, 4 females) aged 40.1 +/- 3.9 for at least three months after their acute event. These survivors were matched for age, sex, body mass index and presence of diabetes mellitus with 98 controls selected from subjects admitted to the same hospital for other reasons. METHODS AND RESULTS: Lipid profiles and APOE polymorphism were determined in both groups. Coronary angiography during hospitalization showed the absence of critical stenosis in 6.6% of the survivors, mono-vessel disease in 57.7%, and multi-vessel disease in 35.5%. The survivors had a higher frequency of smoking, hypertension, family history for coronary artery disease (CAD) and dyslipidemia, and a much greater frequency of 3 or more risk factors than the controls: Odd ratios (OR) 7.4, 95% confidence interval (CI) 2.5-18.6, p = 0.0000. Significant differences were found between the groups for triglycerides (p = 0.000002), total cholesterol (p = 0.003), LDL-cholesterol (p = 0.012), HDL cholesterol (p = 0.0002), apolipoprotein AI (p = 0.00001), and Apolipoprotein B (p = 0.000001). No differences were observed in APOE allele distribution (APOE*4 0.11 vs 0.08, APOE*3 0.86 vs 0.89, APOE*2 0.03 vs 0.03), nor in lipid profile when both higher risk genotype (E3/4, E4/4, E2/4) and lower risk genotype groups (E2/2, E2/3, E3/3) were analysed. OR were calculated as measures of the association of the E4-positive genotypes with AMI. They indicated a non significant increase in risk of AMI when the survivors were compared with the controls (OR 1.78, 95% CI 0.84-3.70, p = 0.13). CONCLUSIONS: This study provides further evidence that conventional coronary risk factors are usually present in young AMI patients. The APOE*4 allele was associated with a 1.8 non-significant increase in the risk of AMI in our group with premature CAD. Comparison with controls showed that the presence of three or more risk factors sharply increased the probability of premature CAD and that hyper-triglyceridemia is an independent risk factor. The data on APOE polymorphism are less certain and a larger study is needed. PMID- 10464785 TI - Modifiable risk factor levels of coronary heart disease survivors in a middle aged workforce. AB - BACKGROUND AND AIM: Coronary heart disease (CHD) is common in New Zealand. Risk factors for CHD are modifiable or non-modifiable. Modifiable risk factor levels of CHD survivors were compared with those without such a history (non-CHD). METHODS AND RESULTS: Participants were from a cross-sectional survey of 5,656 workers aged > or = 40. CHD survivors were 73 general practitioner (GP)-confirmed participants with a history of hospitalisation for CHD. There were no significant differences in mean blood pressure levels between CHD survivors and non-CHD workers after adjusting for age, gender and ethnicity, but current use of antihypertensive medications was higher in CHD survivors (34.2%) than non-CHD workers (8.1%); p < 0.001. CHD survivors had higher, similarly adjusted, mean serum total cholesterol, triglyceride and lower HDL-cholesterol levels, and their reported carbohydrate, fibre, polyunsaturated fat intakes and ratio of polyunsaturated to saturated fat intakes were higher and total fat, saturated fat and monounsaturated fat intakes were lower. CHD survivors ate fewer servings of red meats per month and more servings of fruit, and cereal, and number of cups of milk. Salt added to meals was lower and margarine use higher in CHD survivors. There were no significant differences in the proportions of those who exercised regularly, or were current cigarette smokers. However, more CHD survivors (57.5%) than non-CHD workers (33.1%) were ex-smokers p < 0.001, who had stopped smoking at a higher mean (se) age (41.1 (1.36) vs 37.6 (0.20) years respectively; p = 0.012). CONCLUSIONS: A large proportion of CHD survivors were dyslipidaemic, despite consuming a lower fat, higher fibre and carbohydrate diet. More than 50% of CHD survivors were ex-cigarette smokers, who had given up smoking at a later age than non-CHD workers. These high-risk CHD survivors would benefit from more aggressive measures aimed at correcting their dyslipidaemias. PMID- 10464786 TI - Cholesterol-lowering and vascular reactivity in relation to coronary heart disease. AB - Despite the strong evidence that cholesterol-lowering treatment is effective in the primary and secondary prevention of coronary heart disease, there is a great discrepancy between the results relating to prognosis and the ability of this treatment to induce regression of coronary atheromatous plaques. Since hypercholesterolemia causes a dysfunction in vascular reactivity, improvement can also be ascribed to restoration of vascular relaxation capacity. This conclusion is supported by a wealth of clinical and experimental evidence. PMID- 10464787 TI - Salt-sensitive hypertension: renal and cardiovascular implications. PMID- 10464788 TI - Biotechnology and world food supply. AB - Biotechnology, the use of molecular and cellular tools to genetically modify and improve food supply, will play an increasing and important role in the continuing struggle to produce sufficient food for an ever increasing world population. Many of the approaches will supplement and enhance conventional breeding and also address environmental concerns and help to stabilize food production. This review provides a perspective dealing specifically with crops and three areas of biotechnology, namely, genome analysis with molecular markers, cell- and tissue culture procedures, and the rapid and precise incorporation of genes via transformation. One must remember that biotechnology is still in its infancy, and that approaches will be greatly improved and more efficiently utilized in the future for the betterment of mankind. PMID- 10464789 TI - Green revolution: preparing for the 21st century. AB - In the 1960s there were large-scale concerns about the world's ability to feed itself. However, widespread adoption of "green revolution" technology led to major increases in food-grain production. Between 1966 and 1990, the population of the densely populated low-income countries grew by 80%, but food production more than doubled. The technological advance that led to the dramatic achievements in world food production over the last 30 years was the development of high-yielding varieties of wheat and rice. These varieties are responsive to fertilizer inputs, are lodging resistant, and their yield potential is 2-3 times that of varieties available prior to the green revolution. In addition, these varieties have multiple resistance to diseases and insects and thus have yield stability. The development of irrigation facilities, the availability of inorganic fertilizers, and benign government policies have all facilitated the adoption of green-revolution technology. In the 1990s, the rate of growth in food grain production has been lower than the rate of growth in population. If this trend is not reversed, serious food shortages will occur in the next century. To meet the challenge of feeding 8 billion people by 2020, we have to prepare now and develop the technology for raising farm productivity. We have to develop cereal cultivars with higher yield potential and greater yield stability. We must also develop strategies for integrated nutrient management, integrated pest management, and efficient utilization of water and soil resources. PMID- 10464790 TI - Isolation and characterization of polymorphic microsatellites in Cocos nucifera L. AB - Microsatellites or simple sequence repeats (SSRs) were isolated from coconut (Cocos nucifera) and tested for polymorphism on restricted germplasm. Sequencing of 197 clones from a cv. Tagnanan Tall-enriched genomic library showed that 75% contained a microsatellite, of which 64% were dinucleotide (GA/CT, CA/GT and GC/CG), 6% were trinucleotide, and 30% were compound repeats. Of 41 primer pairs tested on Tagnanan Tall genomic DNA, 38 gave the expected size product, two amplified two loci, and another gave a multilocus pattern. On 20 coconut samples, the 38 SSRs detected 198 alleles (average: 5.2 alleles per microsatellite). Genetic diversity (D = 1 - sigma pi2) values ranged from 0.141 to 0.809. Heterozygotes were present at high frequencies among some dwarf samples. Analysis of similarity matrices based either on shared alleles at each locus (simple matching coefficient) or on allele bands across all loci (Jaccard coefficient) showed similar results. Dwarfs grouped separately from talls and showed less genetic diversity. In a wider test on 40 samples, 8 SSRs detected 64 alleles (average: eight alleles per microsatellite). These results indicate the high potential of microsatellites to detect genetic diversity in coconut germplasm. PMID- 10464791 TI - Isolation and characterization of simple sequence repeat loci in the gray tree frog, Hyla chrysoscelis. AB - A gray tree frog (Hyla chrysoscelis) genomic library was constructed and characterized with regard to the incidence and complexity of simple sequence repeat (SSR) loci. The partial genomic library, containing approximately 10,000 clones with an average-sized insert of 350 bp, was screened with six SSR repeat oligonucleotides (AC, AG, ACG, AGC, AAC, and AAG). Screening identified 31 unique positive clones containing 41 SSR loci. Sequences of tandemly arrayed dinucleotide repeats were more common (36 of 41) than trinucleotide repeats. Twenty-six loci were identified using the AC dinucleotide probe, while 7 loci were identified using the AG dinucleotide probe. An additional 3 AT dinucleotide loci were serendipitously identified. The AT repeats generally comprised the longest dinucleotide repeat loci. The SSR repeat loci reported here should provide potent markers for identity, parentage, and short-lineage determinations in large-scale experiments using gray tree frogs. PMID- 10464792 TI - Chromosomal organization of a sequence related to LTR-like elements of Ty1-copia retrotransposons in Avena species. AB - A repetitive sequence, pAs17, was isolated from Avena strigosa (As genome) and characterized. The insert was 646 bp in length and showed 54% AT content. Databank searches revealed its high homology to the long terminal repeat (LTR) sequences of the specific family of Ty1-copia retrotransposons represented by WIS2-1A and Bare. It was also found to be 70% identical to the LTR domain of the WIS2-1A retroelement of wheat and 67% identical to the Bare-1 retroelement of barley. Southern hybridizations of pAs17 to diploid (A or C genomes), tetraploid (AC genomes), and hexaploid (ACD genomes) oat species revealed that it was absent in the C diploid species. Slot-blot analysis suggested that both diploid and tetraploid oat species contained 1.3 x 10(4) copies, indicating that they are a component of the A-genome chromosomes. The hexaploid species contained 2.4 x 10(4) copies, indicating that they are a component of both A- and D-genome chromosomes. This was confirmed by fluorescent in situ hybridization analyses using pAs17, two ribosomal sequences, and a C-genome specific sequence as probes. Further, the chromosomes involved in three C-A and three C-D intergenomic translocations in Avena murphyi (AC genomes) and Avena sativa cv. Extra Klock (ACD genomes), respectively, were identified. Based on its physical distribution and Southern hybridization patterns, a parental retrotransposon represented by pAs17 appears to have been active at least once during the evolution of the A genome in species of the Avena genus. PMID- 10464793 TI - Cloning and genetic variability of a HindIII repetitive DNA in Acrossocheilus paradoxus (Cyprinidae). AB - Thirty clones of a highly repetitive HindIII fragment of DNA from seven populations of Acrossocheilus paradoxus (Cyprinidae) were isolated and sequenced. The fragment represents a tandemly repeated sequence, with a monomeric unit of 270 bp, amounting to 0.08-0.10% of the fish genome. Higher units of this monomer appear as a ladder in Southern blots. The HindIII satellite DNA family is conserved in three genera of the Cyprinidae. Variation in nucleotide sequences of this repetitive fragment, which is A+T-rich, is distributed both within individuals and among populations. High overall nucleotide divergence (dij = 0.056 +/- 0.001) was detected among clones of the HindIII satellite DNAs of Acrossocheilus paradoxus. Based on the molecular clock hypothesis, the maximum evolutionary rate was estimated to be 5.3 x 10(-7) substitutions per site per year. Lineage sorting may have contributed to the genetic heterogeneity within individuals and populations. Cladistic analyses indicated a closer phylogeographic relationship between populations of the central and south regions in Taiwan. PMID- 10464794 TI - Situs revisited: imaging of the heterotaxy syndrome. AB - Situs anomalies present a diagnostic challenge to radiologists because of the overlapping spectrum of findings commonly seen in asplenia and polysplenia. In a series of 21 patients with a diagnosis of heterotaxy syndrome, all 11 asplenic patients and seven of 10 polysplenic patients had congenital heart disease. Although there was a variety of complex congenital heart disease, the most common type in both patient groups was a common atrioventricular canal. In both groups, the laterality of the aorta and stomach was quite variable, but intestinal malrotation was a constant feature. In 11 asplenic patients, the most frequent findings were a bridging liver (10 cases), absent spleen (10 cases), and left sided inferior vena cava (nine cases). Only seven of these patients had an aorta ipsilateral to the inferior vena cava, contrary to previous thought that this finding was specific for asplenia. In the 10 polysplenic patients, bridging livers were less frequent (five cases), single (six cases) or multiple (four cases) spleens were seen, and azygous continuation with interruption of the inferior vena cava was usually present (eight cases). Although not pathognomonic of polysplenia, inferior vena cava interruption with azygous or hemiazygous continuation is the most common anatomic finding of this condition. Although the terms asplenia and polysplenia are helpful in suggesting the typical anatomy, both syndromes encompass an overlapping spectrum that needs to be described individually and may best be called heterotaxy syndrome. PMID- 10464795 TI - Congenital anomalies of the upper gastrointestinal tract. AB - A wide spectrum of congenital anomalies may affect the upper gastrointestinal tract, including anomalies of the esophagus (e.g., atresia, fistulas, webs, duplications, vascular rings), stomach (e.g., congenital gastric outlet obstruction, duplications), and duodenum (e.g., atresia, annular pancreas, duplications, malrotation). The evaluation of affected patients can require multiple imaging modalities for diagnosis and surgical planning. Radiography is often diagnostic and specific and can usually provide important clues to help determine the optimal diagnostic procedure. Neonates with complete gastric or upper intestinal obstruction do not usually require further radiologic evaluation after radiography: Barium studies are usually contraindicated, and complementary procedures (e.g., ultrasound [US], computed tomography [CT]) are not usually helpful and may even delay surgery, resulting in death. Nevertheless, US has become important in the evaluation of the pediatric gastrointestinal tract and is being used in an increasing number of applications. CT and magnetic resonance imaging are unsuitable for general screening but provide superb anatomic detail and added diagnostic specificity. They are especially useful in demonstrating esophageal duplications and vascular rings as well as associated abnormalities. However, the decision to perform a given imaging examination should be considered carefully to avoid inconvenience or unnecessary radiation exposure to the patient or delays in surgical correction. Quality control programs should be in place to ensure safe, effective radiologic practice through use of up-to-date equipment and good imaging technique. PMID- 10464796 TI - Skeletal complications in pediatric oncology patients. AB - Pediatric oncology patients are at risk for the development of numerous skeletal complications, and radiologic studies are important in the identification and evaluation of these conditions. Methotrexate osteopathy manifests as osteopenia, dense provisional zones of calcification, pathologic fractures, and sharply outlined epiphyses. Hypertrophic osteoarthropathy may occur with nasopharyngeal carcinoma or tumors of the lungs or pleura and manifests as cortical thickening, lamellar periosteal new bone formation, and soft-tissue swelling. Biomechanical abnormalities are often seen at bone scintigraphy in patients who have undergone surgery for bone tumors. Growth plate injury may manifest as marked deformity, sclerotic metaphyseal bands, metaphyseal fraying, and longitudinal striations. Radiation "osteitis" is seen as an initial decrease in bone density with subsequent development of a mixed radiolucent and sclerotic appearance. Ischemic necrosis of the femoral heads is best demonstrated at magnetic resonance (MR) imaging and has low signal intensity on T1-weighted images and a high-signal intensity rim on T2-weighted images. Bone infarcts are seen as well-demarcated, often ring-shaped areas of decreased signal intensity on T1-weighted MR images and as areas of increased signal intensity on short-inversion-time inversion recovery images. Radiographic signs of infection include bone destruction, periosteal new bone formation, and sclerotic changes. Short-inversion-time inversion recovery MR imaging is particularly useful in evaluating posttherapy changes in bone marrow. Osteochondroma may demonstrate a cartilaginous cap at MR imaging, whereas the most important finding in radiation-induced sarcoma is a soft-tissue mass. Radiologists who work with children with cancer need to be familiar with these complications and their imaging appearances. PMID- 10464797 TI - Pseudomembranous colitis: spectrum of imaging findings with clinical and pathologic correlation. AB - Pseudomembranous colitis (PMC) is a potentially life-threatening acute infectious colitis caused by one or more toxins produced by an unopposed proliferation of Clostridium difficile bacteria. PMC is characterized by the presence of elevated, yellow-white plaques forming pseudomembranes on the colonic mucosa. These plaques can be visualized at both pathologic analysis and endoscopy. Plain radiography, contrast enema studies, and computed tomography (CT) are useful in the evaluation of PMC. Plain radiography of the abdomen can demonstrate polypoid mucosal thickening, "thumbprinting" (wide transverse bands associated with haustral fold thickening), or gaseous distention of the colon. A toxic megacolon with distention and occasionally pneumoperitoneum may be seen in the most severe cases of PMC involving perforation. At contrast enema studies, the primary finding in mild cases of PMC is small nodular filling defects representing the mucosal plaques. With more extensive colonic involvement, the plaques are larger and coalesce to form an irregular bowel wall margin. Mural thickening and wide haustral folds caused by intramural edema may also be seen. A contrast enema study is contraindicated in patients with severe PMC due to the danger of perforation. Common CT findings include wall thickening, low-attenuation mural thickening corresponding to mucosal and submucosal edema, the "accordion sign," the "target sign" ("double halo sign"), pericolonic stranding, and ascites. Familiarity with these imaging characteristics may allow early diagnosis and treatment and prevent progression to more serious pathologic conditions. PMID- 10464798 TI - Evaluation of lower back pain with bone scintigraphy and SPECT. AB - Bone scintigraphy and single photon emission computed tomography (SPECT) may be performed for evaluation of lower back pain, especially when a bone abnormality is suspected. Various patterns of tracer activity based on precise identification of the anatomic location of increased uptake may be observed and used to evaluate bones and joints. Lesions centered about the disk space and vertebral body include spondylodiskitis, metastatic disease, vertebral body fracture, and degenerative disease (disk disease, spondylosis deformans). In diskitis, tracer uptake has a vertical orientation. Metastatic involvement should be suspected in solitary lesions evaluated with SPECT when the area of increased uptake extends from the vertebral body into the pedicle. Fractures are seen on planar and SPECT images as a linear, horizontally oriented area of increased uptake centered in the vertebral body. In degenerative disease, increased uptake is centered about the disk space and may be seen in and project beyond the surface of the vertebral body. Lesions of the posterior arch (comprising the pedicle, lamina, and facet joints) include spondylolysis, pedicle lesions, osteoarthritis of the facet joints, and fracture of the transverse process. Scintigraphy may help differentiate long-standing asymptomatic spondylolysis from ongoing disease. In osteoarthritis of the facet joints, SPECT may be used to select patients to be treated with therapeutic injections. Increased uptake in the transverse process most often indicates a fracture, although tumors may also occur in this location. These findings at planar bone scintigraphy and SPECT allow differentiation of common pathologic conditions and can lead to a specific diagnosis. PMID- 10464799 TI - SPECT and PET in the evaluation of coronary artery disease. AB - Cardiac positron emission tomography (PET) is an accurate method for assessing myocardial perfusion and metabolism in the evaluation of coronary heart disease. PET allows more accurate detection of myocardial ischemia than single photon emission tomography (SPECT). In addition, PET has higher spatial resolution and allows attenuation correction and the quantification of various physiologic parameters. PET with 2-(fluorine-18) fluoro-2-deoxy-D-glucose is considered the standard of reference for predicting improvement in regional or global left ventricular function after revascularization by identifying hibernating viable myocardium that shows diminished perfusion and preserved metabolism. Other less commonly used clinical applications of cardiac PET include assessment of myocardial oxygen consumption and fatty acid metabolism. The use of PET in myocardial imaging is expected to increase in the near future with the regional distribution of positron-emitting radiotracers and the emergence of relatively low-cost PET systems. PMID- 10464800 TI - Focal intracavitary masses recognized with the hyperechoic line sign at endovaginal US and characterized with hysterosonography. AB - In 42 women, endovaginal ultrasonography (US) was performed because of an abnormal central endometrial complex. Retrospective review of the endovaginal US scans and subsequently obtained hysterosonograms with histopathologic correlation showed a hyperechoic line surrounding the central endometrial complex in 25 patients, all of whom had focal intracavitary processes. Cystic spaces within the central endometrial complex in 17 of these patients reflected polyps; this finding was associated with a hyperechoic line in 15. At hysterosonography, all polyps (n = 29) had a narrow attachment to the endometrium. All five fibroids were broadly attached and had a rim of overlying endometrium. These focal abnormalities were easily distinguished from global endometrial processes amenable to a blind biopsy procedure. This initial experience showed that a hyperechoic line circumscribing the central endometrial complex at endovaginal US allows prediction of focal intracavitary disease, particularly when the line is associated with cystic spaces within the central endometrial complex. The presence of a hyperechoic line can be used to direct the patient to a visually guided hysteroscopic procedure rather than a potentially unsuccessful blind biopsy procedure. Alternatively, absence of a hyperechoic line can prompt management appropriately directed by hysterosonographic characterization of the uterine contents as focal or diffuse. PMID- 10464801 TI - Staging of early endometrial carcinoma: assessment with T2-weighted and gadolinium-enhanced T1-weighted MR imaging. AB - This study evaluated the usefulness of T2-weighted and gadolinium-enhanced T1 weighted magnetic resonance (MR) images correlated with patients' menopausal status in assessing the depth of myometrial invasion in stage I endometrial carcinoma. MR images of 46 patients with stage I endometrial carcinoma were retrospectively reviewed. Twenty-five patients were premenopausal, and 21 were postmenopausal. The staging accuracy without regard to menopausal status was 59% for T2-weighted images and 61% for gadolinium-enhanced T1-weighted images. However, when staging accuracy was evaluated separately in the premenopausal and postmenopausal patient groups, T2-weighted imaging had an accuracy of 80% in the premenopausal group and gadolinium-enhanced T1-weighted imaging had an accuracy of 81% in the postmenopausal group. Therefore, T2-weighted imaging was more accurate in premenopausal patients and gadolinium-enhanced T1-weighted imaging was more accurate in postmenopausal patients. The overall accuracy of staging with MR imaging improved to 80% when patients' menopausal status was considered. Therefore, menopausal status should be considered when T2-weighted and gadolinium enhanced T1-weighted MR images are used to stage early endometrial carcinoma. PMID- 10464802 TI - Imaging of nontraumatic hemorrhage of the adrenal gland. AB - Nontraumatic hemorrhage of the adrenal gland is uncommon. The causes of such hemorrhage can be classified into five categories: (a) stress, (b) hemorrhagic diathesis or coagulopathy, (c) neonatal stress, (d) underlying adrenal tumors, and (e) idiopathic disease. Computed tomography (CT), ultrasonography (US), and magnetic resonance (MR) imaging play an important role in diagnosis and management. CT is the modality of choice for evaluation of adrenal hemorrhage in a patient with a history of stress or a hemorrhagic diathesis or coagulopathy (anticoagulant therapy). CT may yield the first clue to the diagnosis of adrenal insufficiency secondary to bilateral massive adrenal hemorrhage; such insufficiency is rare but life threatening. US is the modality of choice for evaluation of neonatal hematoma, and MR imaging is helpful for further characterization. MR imaging is also useful in the diagnosis of coexistent renal vein thrombosis. When an adrenal abscess is suspected, percutaneous aspiration and drainage under imaging guidance should be performed. Hemorrhage into an adrenal cyst or tumor can cause acute onset of symptoms and signs in a patient without discernible risk factors for adrenal hemorrhage. A hemorrhagic adrenal tumor should be suspected when CT or MR imaging reveals a hemorrhagic adrenal mass of heterogeneous attenuation or signal intensity that demonstrates enhancement. PMID- 10464803 TI - Pulmonary parenchymal manifestations of mitral valve disease. AB - Pulmonary parenchymal manifestations of mitral valve disease are the result of either pulmonary venous hypertension in mitral stenosis or abnormal regurgitant flow into pulmonary veins in mitral insufficiency. Typical radiographic findings in mitral stenosis include pulmonary vascular cephalization; interstitial, perivascular, and occasionally alveolar pulmonary edema; diffuse alveolar hemorrhage; hemosiderosis; and pulmonary ossification. Signs of interstitial pulmonary edema are frequently visible and include septal lines. Radiographic findings in diffuse alveolar hemorrhage consist of diffuse, confluent acinar or ground-glass areas of increased opacity, often sparing the peripheral parenchyma and creating the so-called window frame effect. Hemosiderosis is characterized by small, ill-defined nodules or by coarse reticular areas of increased opacity with a bias for the middle and lower lung regions. Ossification manifests as densely calcified, 1-5-mm nodules, mainly in the middle and lower lungs, with a tendency for confluence and the occasional presence of trabeculae. Imaging findings in mitral regurgitation depend on the acuteness of the disease. The most common parenchymal manifestations of acute mitral regurgitation are symmetric alveolar and interstitial pulmonary edema with indistinct, engorged pulmonary vessels and cephalized blood flow. Familiarity with these manifestations can expedite diagnosis, particularly in rare cases of unsuspected mitral valve disease. PMID- 10464805 TI - Adrenocortical neoplasms in children: radiologic-pathologic correlation. AB - Primary neoplasms of the adrenal cortex are rare in children and differ significantly in epidemiology, clinical characteristics, and biologic features from their counterparts in adults. In children, the inclusive term adrenocortical neoplasm is applied because adrenal adenoma and adrenal carcinoma may be difficult to distinguish histopathologically. Pediatric adrenocortical neoplasms typically occur before 5 years of age, affect young girls more commonly than boys, and are associated with hemihypertrophy and Beckwith-Wiedemann and Li Fraumeni syndromes. Most children with an adrenocortical neoplasm present with signs and symptoms of endocrine abnormality, including virilization and Cushing syndrome. Cross-sectional imaging studies typically demonstrate a large, circumscribed, predominantly solid suprarenal mass with variable heterogeneity due to hemorrhage and necrosis. Calcification is not uncommon. Local invasion and metastases to the lungs, liver, and regional lymph nodes may be present at diagnosis. When friable tumor thrombus extends into the inferior vena cava, it poses a high risk of pulmonary embolization. The finding of increased retroperitoneal fat due to hypercortisolism on computed tomographic and magnetic resonance images of children with an adrenal mass favors the diagnosis of adrenocortical neoplasm. Surgical resection is the mainstay of therapy, with chemotherapy used for patients with metastases or persistent elevated hormone levels following surgery. Patients younger than 5 years with aggressive adrenocortical neoplasms fare better than older children. PMID- 10464804 TI - Contrast-enhanced three-dimensional MR portography. AB - Three-dimensional (3D) magnetic resonance (MR) portography with contrast material enhancement is a fast means of evaluating the portal venous system that has some advantages over currently used modalities, such as digital subtraction angiography, helical computed tomography, ultrasonography, and nonenhanced MR angiography with time-of-flight and phase-contrast techniques. With contrast enhanced 3D MR portography, a first-pass study of the mesenteric vasculature is performed after rapid bolus injection of gadopentetate dimeglumine; a 3D fast field echo sequence is used, which can demonstrate the intrahepatic and extrahepatic portal venous system clearly. Repeated sequences after administration of gadopentetate dimeglumine allow separate demonstration of the splanchnic arteries and portomesenteric veins. The images are reconstructed by means of maximum-intensity projection postprocessing, and a subtraction technique can be used to eliminate arterial enhancement and demonstrate portosystemic shunts. The coronal source images simultaneously demonstrate parenchymal lesions of the liver, pancreas, biliary tract, and spleen. This technique is clinically indicated in portosystemic shunt, portal vein thrombosis, hepatocellular carcinoma, pancreatobiliary tumor, hepatic vein obstruction, differentiation of splanchnic arterial from portal venous disease, and gastrointestinal hemorrhage. Its limitations include allergic reactions to contrast media, inappropriate positioning of the 3D acquisition slab, respiratory motion artifacts, and pseudodissection. PMID- 10464806 TI - Cardiac MR imaging: a guide for the beginner. AB - The complex motion of the heart during contraction is a serious challenge to the diagnostic radiologist and to the capabilities of the magnetic resonance (MR) imaging unit itself, but electrocardiographic (ECG) gating "stops" motion and thus allows acquisition of diagnostic-quality images. Attention to placement of chest-wall ECG electrodes and the course of ECG leads from the patient will maximize the quality of the gating signal and result in better-quality images. Before commencing the MR imaging examination, the clinical questions for the examination must be identified so that a clinical protocol can be applied to acquire relevant morphologic and physiologic data. In addition to the standard orthogonal views (axial, coronal, and sagittal), oblique and complex sections parallel and orthogonal to intrinsic cardiac axes may be necessary to portray the relevant anatomy to best advantage. Construction of these views requires an understanding of basic normal and pathologic cardiac anatomy. If care is taken in the planning and execution of the MR imaging examination, the radiologist will be able to exploit this exciting technology to its full noninvasive potential. PMID- 10464807 TI - Radiation safety considerations for diagnostic radiology personnel. AB - In radiation protection, the guiding philosophy is ALARA (as low as reasonably achievable), and states have regulatory authority. Dose limits are in part based on effective dose equivalent and differences in tissue sensitivities. In diagnostic radiology, the main source of occupational dose is scattered radiation from the patient--particularly from fluoroscopically guided procedures. Personnel stand near patients for long times, and angulated geometries with C-arm equipment may result in high personnel doses from backscatter. For all procedures, judicious applications of time, distance, and shielding affect dose. Appropriate use includes collimating properly, optimizing beam-on time, minimizing distances between image intensifier and patient, ensuring sufficient distance between patient and x-ray tube, and optimizing exposure rates for image quality and dose. Although dose limits typically regulate maximum whole-body dose, protective clothing worn by fluoroscopists reduces personnel risks; weighting factors can be applied to estimate effective dose equivalent. Pregnant personnel have lower limits, which apply only with voluntary declaration of pregnancy. With appropriate precautions, fetal doses can typically remain within recommended limits without changes in occupational tasks. Radiation workers in each state must ensure that regulations are appropriate. Then, for protection of both employee and employers, the rules can and must be followed. PMID- 10464808 TI - Effect of field strength on MR images: comparison of the same subject at 0.5, 1.0, and 1.5 T. AB - To assess the effect of field strength on magnetic resonance (MR) images, the same healthy subject was imaged at three field strengths: 0.5, 1.0, and 1.5 T. Imaging was performed with three similarly equipped MR imagers of the same generation and from the same manufacturer. The same imaging sequences were used with identical parameters and without repetition time correction for field strength. Imaging was performed in four anatomic locations: the brain, lumbar spine, knee, and abdomen. Quantitative image analysis involved calculation of signal-to-noise ratio, contrast-to-noise ratio, and relative contrast; qualitative image analysis was performed by four readers blinded to field strength. The results of all of the examinations were considered to be of diagnostic value. In general, signal-to-noise ratio and contrast-to-noise ratio were lowest at 0.5 T and highest at 1.5 T; relative contrast was not related to field strength. At qualitative analysis, images obtained at 1.0 and 1.5 T were superior to images obtained at 0.5 T; qualitative differences were less important in locations where there is motion or high magnetic susceptibility differences between tissues (e.g., the spine and abdomen). However, excellent image quality was obtained with all three field strengths. PMID- 10464809 TI - Consulting with radiologists outside the hospital by using Java. AB - A Java-based teleradiology system that makes use of the Internet has been developed. Using this system, an on-call, off-duty radiologist can make diagnoses and perform consultations easily by reviewing the transferred images at home. The image accessibility of the system allows a hospital with such a system to assist an affiliated rural hospital without a full-time radiologist. The system consists of three components: the image server subsystem, the database server subsystem, and the client subsystem. All client subsystems of the system are designed to be symmetric. Users may retrieve images, manipulate them, and perform remote consultations. In addition, a user may add annotations to an image area of interest. Screen synchronization is achieved by means of the command-passing technique and local command execution to reduce the network bandwidth and transmission demands; user interaction is achieved by means of a synchronized indicator for pointing out areas of interest and dialog windows for conversation. Because Java programs can run on heterogeneous platforms, the need for system maintenance and user training is minimized. Although the drawback of Internet bandwidth still exists, the system enables platform-independent teleradiology via the Internet and allows easy and cost-effective remote consultation. PMID- 10464810 TI - Strongyloides stercoralis hyperinfection. PMID- 10464811 TI - General case of the day. Mesenteric panniculitis with extensive inflammatory involvement of the peritoneum and intraperitoneal structures. PMID- 10464812 TI - Breast imaging case of the day. Pseudoangiomatous stromal hyperplasia. PMID- 10464813 TI - US case of the day. Placenta increta. PMID- 10464814 TI - Pediatric case of the day. Klippel-Trenaunay syndrome. PMID- 10464815 TI - Neuroradiology case of the day. Superior sagittal sinus thrombosis and infarcts. PMID- 10464816 TI - Gastrointestinal case of the day. Chronic ileocolocolic intussusception secondary to a mobile cecum and a benign fibrovascular mass. PMID- 10464817 TI - Pneumocystis carinii pneumonia in HIV-infected patients: effect of steroid therapy on surfactant level. AB - Previous studies have suggested alterations in pulmonary surfactant lipid in the setting of Pneumocystis carinii pneumonia in HIV-infected patients. Because pulmonary surfactant lipid is composed of a variety of lipid products and because other phospholipids might be present in bronchoalveolar lavage (BAL) lipid determinations, a single molecular species of phospholipid which comprises a substantial portion of the surfactant lipid fraction, dipalmitoyl phosphatidylcholine (DPPC), was measured by capillary column gas chromatography in BAL samples taken at the time of the diagnosis of P. carinii pneumonia, and 10 days after treatment for P. carinii pneumonia. DPPC was measured at day 0 and day 10 in seven patients who had been randomized to receive methylprednisolone adjuvant therapy for P. carinii pneumonia and in six patients who had been randomized to not receive methylprednisolone therapy. The level of DPPC in BAL from all patients at day 0 was 0.49 +/- 0.06 microgram ml-1 BAL. This level is significantly lower that the level of DPPC determined in BAL from five normal volunteers 2.48 +/- 0.40 micrograms ml-1. At day 0, the BAL level of DPPC in patients treated with methylprednisolone was not different from the BAL level of DPPC in patients not treated with methylprednisolone. By day 10 of therapy for P. carinii pneumonia, BAL levels of DPPC in all patients had increased to 1.05 +/- 0.19 micrograms ml-1 BAL. At day 10 DPPC levels in the methylprednisolone treated group were not different from the group not treated with methylprednisolone. We conclude that in HIV-infected patients, lung surfactant lipid is reduced in the setting of P. carinii pneumonia. The lipid levels return toward normal levels with treatment. Adjuvant therapy with corticosteroids does not alter the rate of recovery of surfactant lipid levels at least after 10 days of therapy. PMID- 10464818 TI - Delayed chest radiographs and the diagnosis of pneumothorax following CT-guided fine needle aspiration of pulmonary lesions. AB - We prospectively studied 64 consecutive patients on whom computerized tomography guided fine needle aspirations (CT-FNA) of pulmonary lesions were performed to determine whether obtaining a delayed chest radiography (CXR) after CT-FNA is helpful in the diagnosis of post-procedure pneumothorax (PTX). Two of the 64 patients developed a delayed PTX. Only one of the patients with a delayed PTX required chest tube drainage. If patients have no evidence of a PTX immediately after a CT-FNA by CT scan then a delayed CXR adds little to care of these patients. PMID- 10464819 TI - The effect of gestational parity on FEV1 in a group of healthy volunteer women. AB - In the past, studies utilizing within-subject comparisons of small groups of pregnant women showed that forced expiratory volume in 1 s (FEV1) remained essentially unchanged during pregnancy. However, one of the findings from an epidemiological study was that women with greater number of children experienced a faster decline of FEV1. The aim of this study was to examine the effect of parity on FEV1 in a group of healthy volunteer women. To this end, cross sectional multiple regression analyses of data from 397 healthy women participants in the Baltimore Longitudinal Study of Aging (BLSA) with a mean (range) age of 47.7 (18-92) years were performed. Similar analyses were done using the younger (50 years or less) and the older (> 50 years) subgroups. After controlling for age, height, weight, and smoking, parity as a dichotomous variable was associated with a higher FEV1 in women of child-bearing age (0.139 1; P = 0.02) but not in the older women. There was a modest link with the number of children (P = 0.05), with the first child possibly having the greatest effect on FEV1. We could not account for the effect of parity on FEV1 by the educational level, occupation, health status of the women, or by the presence of a cohort effect. Thus the nulliparous state is associated with lower FEV1 in this group of healthy adult women of child-bearing age. PMID- 10464820 TI - Increased content of thiobarbituric acid-reactive substances and hydrogen peroxide in the expired breath condensate of patients with stable chronic obstructive pulmonary disease: no significant effect of cigarette smoking. AB - The imbalance between oxidants and antioxidants is known to play an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). Cigarette smoking is the most frequent factor responsible for development of COPD by leading to oxidant overload in the lower airways, due to presence of its own oxidants and to recruitment and activation of pulmonary phagocytes. We aimed to determine whether (1) patients with stable COPD have higher thiobarbituric acid reactive substances (TBARs, an end-product of lipid peroxidation) and H2O2 levels in expired breath condensate than healthy subjects who have never smoked; (2) COPD subjects who are current smokers exhale more TBARs and H2O2 than COPD ex smokers and those who have never smoked; and (3) concentration of TBARs correlates with H2O2 levels in the breath condensate of COPD patients. The TBAR and H2O2 content in expired breath condensate of 17 healthy nonsmoking subjects and 44 patients (11 current smokers, 20 ex-smokers and 13 who had never smoked) with stable COPD [forced expiratory volume in 1 s (FEV1) 63.3 +/- 16.3% and FEV1 reversibility 5.2 +/- 4.3% predicted value] was measured spectrofluorimetrically by the thiobarbituric acid and homovanillic acid methods, respectively. The mean concentrations of TBARs and H2O2 in the expired breath condensate of COPD subjects were 12 (0.48-0.86 microM vs. 0.04 +/- 0.14 microM; P < 0.05) and 10 times (0.48 +/- 0.67 microM vs. 0.05 +/- 0.07 microM; P < 0.005) higher than in healthy controls. Current smokers with COPD did not exhale more H2O2 than COPD ex smokers and those who had never smoked. TBARs levels shared only a tendency to be higher in the breath condensate of smoking COPD subjects than in that of ex smokers (0.92 +/- 1.49 microM vs. 0.35 +/- 0.44 microM) and of COPD subjects who had never smoked (0.92 +/- 1.49 microM vs. 0.30 +/- 0.53 microM). No correlation was found between TBAR and H2O2 levels in the whole COPD group. These variables did not correlate with cigarette smoking status and the time from smoking cessation. Subjects with stable COPD exhibit increased lipid peroxidation and H2O2 generation in the airways. Current cigarette smoking does not distinguish COPD subjects with respect to TBARs and H2O2 exhalation. PMID- 10464821 TI - Are doctors still failing to assess and treat asthma attacks? An audit of the management of acute attacks in a health district. AB - This audit aimed to observe the management of acute asthma by primary and secondary care within a Health District. Asthma attacks occurring during the first 6 weeks of 1996 to patients between the ages of 3 and 74 years in Canterbury and Thanet District were notified by general practitioners, out-of hours co-operatives and hospitals. Data were obtained retrospectively from the patient records. A total of 378 episodes was registered: 342 (90%) to primary care. Of these 234 (76% of patients aged 6 years or over) had a peak flow recorded; 114 (30%) were given emergency bronchodilation: oxygen was not used in primary care; 204 (54%) were given systemic steroids; and 43 (11%) were referred for hospital care of whom 36 were admitted. Of the attacks, 212 (69% of the patients aged 6 years or over) could be classified by percentage predicted peak flow and management compared to the Guidelines published by the British Thoracic Society. Twenty-eight patients presented with 'life-threatening' asthma: 20 (71%) were given emergency bronchodilation; oxygen was used in only two; 24 (86%) were given systemic steroids; and six (21%) were referred for admission. In their confidential enquiry into the asthma deaths the British Thoracic Society identified a failure to appreciate the severity of the attack, resulting in inadequate emergency treatment and delay in referring to hospital. These data suggest that, 15 years later, these problems may still exist. PMID- 10464822 TI - The cost-effectiveness of inhaled fluticasone propionate and budesonide in the treatment of asthma in adults and children. AB - Inhaled corticosteroids form the mainstay of the treatment and management of asthma and the results of a meta-analysis comparing two of the most frequently prescribed inhaled corticosteroids, fluticasone propionate and budesonide, administered in a clinically equivalent 1:2 dose ratio to 1980 patients with asthma, demonstrated that fluticasone propionate had an improved efficacy:safety ratio. However, limited data are available on the relative economic benefits of fluticasone propionate and budesonide. The database for clinically relevant parameters, for which the efficacy:safety meta-analysis had demonstrated statistical significance between the two corticosteroids, was used for this pharmacoeconomic analysis. Treatment with fluticasone propionate was more cost effective than budesonide with respect to improvement in morning peak expiratory flow rate, successfully treated weeks, symptom-free days, symptom-free 24 h and episode-free days. The costs of treatment for fluticasone propionate and budesonide were 7.78 Pounds per week and 12.33 Pounds per week, respectively. The main contributing factor to the higher costs of budesonide was the higher cost of health care contacts, which were 4.53 Pounds per week for budesonide and 0.57 Pounds per week for fluticasone propionate. The pharmacoeconomic difference increased in favour of fluticasone propionate as the criteria for success were made more stringent. These results demonstrate that, for asthma patients requiring modification of therapy treatment with fluticasone propionate is more effective and also cheaper, in terms of overall health-care costs, than treatment with budesonide. PMID- 10464823 TI - Familial sarcoidosis in Finland and Hokkaido, Japan--a comparative study. AB - Two or more cases of sarcoidosis in one family is not unusual. To compare the frequencies of familial sarcoidosis in Finland and Hokkaido, Japan, and to analyse the type of associations reported, we collected data on all patients visiting hospitals for sarcoidosis in 1984 in Finland (1378 patients) and Hokkaido (208 patients), including information about familial sarcoidosis. We also analysed the familial cases seen among 571 sarcoidosis patients diagnosed at the Mjolbolsta hospital in Finland from 1955 to 1987 and among 686 Japanese patients seen in Sapporo from 1964 to 1988. In 1984, 50 sarcoidosis patients visiting Finnish hospitals and nine sarcoidosis patients in Hokkaido reported as familial cases. Of the sarcoidosis patients seen in Finland at the Mjolbolsta hospital in 1955-1987, 27 had a family member with the same disease, while this number was 20 in the Sapporo hospital in 1964-1988. Those surveys give a prevalence of familial sarcoidosis in Finland of 3.6-4.7% and in Hokkaido of 2.9 4.3%. Among familial cases, the dominating relationships were sister-brother and mother-child relationships. PMID- 10464824 TI - The role of codeine phosphate premedication in fibre-optic bronchoscopy under insufficient local anaesthesia and midazolam sedation. AB - Midazolam is widely used as a sedative agent to produce amnesia in patients undergoing fibre-optic bronchoscopy. However, if a patient does not receive sufficient local anaesthesia, continuous severe cough and physical movement may interrupt the procedure and reduce its safety. We therefore examined whether codeine phosphate is a useful premedication for bronchoscopy. The study design was a randomized comparison between codeine phosphate and a placebo in patients undergoing light local anaesthesia and midazolam sedation. We used low dose local anaesthesia (5 ml of nebulized 2% xylocaine) on the assumption of insufficient local anaesthesia. Patients were allocated to receive codeine phosphate 0.4 mg kg 1 or a saline placebo 60 min before they were sedated with i.v. midazolam. If the patients exhibited severe cough during bronchoscopy, intrabronchial supplemental local anaesthesia (2% xylocaine solution in 1 ml increments) was instilled via a bronchoscope to the trachea and segmental bronchi to suppress the cough. The dose of supplemental xylocaine was assessed and the requirements were significantly lower in the codeine group compared to the placebo group: 36.4 +/- 10.2 mg vs. 95.1 +/- 24.6 mg, respectively. After bronchoscopy, patients were interviewed by a doctor to assess their willingness to undergo a repeat procedure if one was clinically indicated, but no significant difference was observed between the two groups. If local anaesthesia is insufficient, midazolam together with codeine phosphate premedication is useful for both the patient and the bronchoscopist. PMID- 10464825 TI - beta 2-agonist-induced inhibition of neutrophil chemotaxis is not associated with modification of LFA-1 and Mac-1 expression or with impairment of polymorphonuclear leukocyte antibacterial activity. AB - Patients with chronic obstructive lung disorders often show increased susceptibility to airway infections. As beta 2-adrenoceptor agonists, in addition to reversing the contractile response of bronchial smooth muscles, may inhibit a variety of inflammatory and immuno-effector cell functions, it is possible that these drugs interfere with host defence mechanisms. The present study was designed to test in vitro whether fenoterol, a short-acting beta 2-adrenoceptor agonist, could modify human blood neutrophil recruitment and antimicrobial activity. Pre-exposure to fenoterol significantly reduced neutrophil migration towards the complement component C5a, at concentrations ranging from 10(-7) M to 10(-5) M, or towards lipopolysaccharide, at a concentration of 10(-5) M (P < 0.05, each comparison). In contrast, the drug (10(-8)-10(-5) M) did not significantly modify the increased expression of lymphocyte function-associated antigen (LFA-1, i.e. CD11a/CD18) the macrophage antigen-1 (Mac-1, i.e. CD11b/CD18) induced by N-formylmethionylleucylphenylalanine (fMLP) (P > 0.05, each comparison). Finally, incubation of neutrophils with fenoterol (10(-8)-10( 5) M) did not significantly influence phagocytosis or intracellular killing of bacteria (Staphylococcus aureus) or H2O2 release induced by tetradecanoyl-phorbol acetate (P > 0.1 for each comparison). These results suggest that short-acting beta 2-adrenoceptor agonists, such as fenoterol, are able partially to reduce neutrophil recruitment in the airways without interfering with the processes involved in phagocytic activity against bacteria. PMID- 10464826 TI - Improved targeting of beclomethasone diproprionate (250 micrograms metered dose inhaler) to the lungs of asthmatics with the Spacehaler. AB - The Spacehaler (Evans Medical Ltd, Leatherhead, U.K.) is a new, compact, inhaler device containing the same aerosol canister as a conventional metered dose inhaler (MDI). However, the design of the Spacehaler has been shown to reduce the velocity of the aerosol, thus reducing the proportion of non-respirable particles delivered to the patient. This study compared radioaerosol deposition patterns following inhalation of 250 micrograms of beclomethasone dipropionate from the Spacehaler and a conventional MDI (Beclazone, Norton Health Care, Harlow, U.K.). After rigorous in vitro validation of the radiolabelling technique, 12 asthmatic subjects (seven men aged 20-69 years, mean baseline FEV1 2.59 1 (SD 0.55 1) received one dose of 99mTc-labelled beclomethasone dipropionate 250 micrograms via either a Spacehaler or MDI on each of two study days in a randomized cross over manner. All subjects had been taught the required inhalation technique before the dose was administered. Inhalation details were recorded using a spirometer connected in series with the device. Lung and oropharyngeal depositions were measured by gamma scintigraphy. The mean percentage of the metered dose deposited in the lungs was 23.0% (SD 8.3%) for the Spacehaler and 12.8% (SD 6.8%) for the MDI (P < 0.01). However, there was no significant difference in the distribution patterns within the lungs between the two devices. Oropharyngeal deposition was significantly lower (P < 0.01) for the Spacehaler than for the MDI [mean (SD) 27.9% (16.4%) and 73.6% (8.7%), respectively] whilst the percentage of the metered dose remaining on the Spacehaler actuator was significantly greater than that on the MDI actuator [mean (SD) 48.0% (11.8%) and 12.4% (8.5%) respectively, P < 0.01]. There was evidence from the inhalation recordings that some patients experienced the 'cold Freon effect' whilst using the metered dose inhaler which may have contributed to the lower lung deposition seen with this device. This study demonstrates that the proportion of a 250 micrograms dose of beclomethasone dipropionate that is delivered to the lungs is significantly greater with the Spacehaler than the MDI. The Spacehaler also reduces the proportion of the does that is deposited in the oropharynx to less than half that observed with the MDI, and reduces the total dose of drug received by the patient. PMID- 10464827 TI - Autologous blood-patch pleurodesis for secondary spontaneous pneumothorax with persistent air leak. PMID- 10464828 TI - High-frequency oscillatory ventilation in RSV bronchiolitis patients. PMID- 10464829 TI - A comparison of the efficacy of long-acting beta 2-agonists: eformoterol via Turbohaler and salmeterol via pressurized metered dose inhaler or Accuhaler, in mild to moderate asthmatics. Force Research Group. AB - Four hundred and sixty nine patients were randomized to receive either 12 micrograms bd of eformoterol (Oxis, Astra Pharmaceuticals Ltd., Kings Langley, U.K.) delivered via Turbohaler or 50 micrograms bd salmeterol (Serevent, Glaxo Wellcome Ltd., Uxbridge, U.K.) via either the Accuhaler (Glaxo-Wellcome Ltd.) or pressurized metered dose inhaler (pMDI, Glaxo-Wellcome Ltd.) for 8 weeks. This was followed by a 4-week cross-over period when patients who had received salmeterol in the previous 8 weeks were given eformoterol and patients who had received eformoterol were given either salmeterol via the Accuhaler or pMDI to assess patient device and treatment preference. For the primary efficacy variable, the increase in peak expiratory flow (PEF) rate from run-in to 8 weeks, similar significant improvements were seen in all three treatment groups. Eformoterol Turbohaler (FT) achieved a greater increase in morning PEF than salmeterol Accuhaler (SA) from randomisation to 4 weeks; the increase shown in the eformoterol Turbohaler group was 28.9 1 min-1 compared to 19.91 min-1 for the salmeterol Accuhaler group. The addition of eformoterol Turbohaler 12 micrograms bd, to patients' existing asthma therapy was found to have a significantly more beneficial effect on the severity of patients' daytime asthma symptoms than had salmeterol Accuhaler 50 micrograms bd (P = 0.014). Eformoterol Turbohaler reduced the severity of daytime asthma symptoms by 42% after only 4 weeks of treatment. The patients in the eformoterol Turbohaler treated group experienced a higher percentage of days when they were symptom-free and did not use their short-acting bronchodilator to relieve symptoms (32.8, 24.1 and 28.0% in the FT, SA and SM groups, respectively). At 8 weeks there were no significant differences in any of these variables between the three groups. Patients in all the treatment groups gained an additional 1-1.5 nights undisturbed by asthma per week. The changes in sleep disturbance were not significantly different between the three treatment groups. In addition to the therapeutic benefits provided by eformoterol Turbohaler the device (Turbohaler) was the significant preference of patients given both Turbohaler and pMDI (P = 0.0168) and was also considered to be significantly more convenient to carry around than the Accuhaler (P < 0.0001). No other differences were found between the three devices. The results of this study demonstrate that the addition of a long-acting beta 2-agonist is an effective tool for achieving the goals of asthma treatment. Eformoterol via the Turbohaler is at least as effective as salmeterol via either the Accuhaler or the pMDI in achieving these goals. PMID- 10464830 TI - Regular albuterol or nedocromil sodium--effects on airway subepithelial tenascin in asthma. AB - Both albuterol and nedocromil sodium have been recognized to possess certain anti inflammatory properties. However, there are no data on the impact of these drugs on the pathophysiology of the bronchial extracellular matrix in asthma characterized by enhanced tenascin (Tn) expression, known to occur proportional to the severity of asthma. This paper reports data from a morphometric study on the effects of regular treatment with inhaled albuterol or nedocromil sodium on the extent of bronchial subepithelial deposition of Tn, collagen types III, IV, and VII and mucosal infiltration with macrophages. Thirty-two patients (14 women) with chronic asthma, aged 38.7 years (median) with a median forced expiratory volume in 1 sec (FEV1) of 74.4% predicted, were selected to undergo fibre-optic bronchoscopy with bronchial biopsies before and after 12 weeks of treatment with either inhaled albuterol 0.2 mg or nedocromil sodium 4 mg four times daily according to a double-blind protocol. Cryostat sections of the biopsy specimens were studied by indirect immunostaining techniques using monoclonal antibodies and computer-assisted quantitative image analysis. Albuterol treatment significantly reduced the median thickness of subepithelial Tn expression from 9.7 to 6.3 microns (P = 0.023) and macrophage numbers in the epithelium (P = 0.034), lamina propria (P = 0.039) and entire mucosa (P = 0.033), whereas nedocromil sodium had no effect. Expression of the collagen types was not affected by either treatment. There was no identifiable statistical difference between the two treatments for any of the outcome variables measured. Nevertheless, the results demonstrate that even a short-acting beta 2-agonist may exert anti-inflammatory potential sufficient to interfere with the basic mechanisms of asthma as shown by reduction of subepithelial Tn content and mucosal macrophage count. PMID- 10464831 TI - Clinical value and cost of a respiratory sleep-related breathing disorders screening service for snorers referred to a District General Hospital ENT department. AB - Sleep-related breathing disorders and snoring often co-exist in the community. We hypothesized that a significant proportion of patients referred from primary care to ENT surgeons for management of snoring might have significant sleep-related breathing disorders requiring medical management. The Respiratory Medicine Department at Whipps Cross Hospital, London, U.K. screened all such referrals using sleep questionnaires, overnight oximetry and diagnostic sleep studies where necessary as recommended by the Royal College of Physicians of London. Over 38 months, 115 patients were screened, of whom 43 (38%) had clinically significant sleep-disordered breathing. One-third were established on nasal continuous positive airway pressure ventilation and the remainder were mainly offered conservative treatment. The cost of the screening service is estimated at 14,000 Pounds for the initial year. The savings to the ENT service and the possible long term benefits to the patients identified as having sleep-disordered breathing balance this. We conclude that screening all referred snorers for sleep disordered breathing using a simple protocol identifies a significant number requiring medical management at a relatively low cost to the service provider. PMID- 10464832 TI - Safety and cellular assessment of bronchial brushing in airway diseases. AB - Bronchial brushing is a useful method for morphological and functional studies of bronchial epithelial cells (BECs) in various diseases. This technique has been found to be generally safe, but its safety in asthma and chronic bronchitis has not been fully assessed. The purpose of this study was to determine 1, whether bronchial brushing is a safe method in asthmatic and chronic bronchitis patients of differing severity and 2, to characterize the BECs obtained in terms of number, viability and purity. We evaluated 25 asthmatics of variable severity, 19 chronic bronchitis patients and 26 normal volunteers. Bronchoscopy and bronchial brushing were performed in a standardized manner by the same investigator. Safety was assessed by clinical follow-up of all subjects; continuous monitoring of arterial oxygen saturation during the procedure with a digital oximeter was carried out in a subsample of subjects. No complications were observed clinically during the procedure. There was a minimal fall in arterial oxygen saturation without a significant difference between the three groups of subjects. A consistent number of BECs was recovered and their viability, assessed by the trypan blue exclusion test, in asthmatics and chronic bronchitis patients was significantly lower than in controls (P < 0.05). Bronchial brushing is well tolerated and may be a valuable method of obtaining BECs in asthmatic and chronic bronchitis patients. PMID- 10464833 TI - Performance of a short lung-specific health status measure in outpatients with chronic obstructive pulmonary disease. AB - The objective of this study was to assess the performance of a lung-specific health status measure in patients with chronic obstructive pulmonary disease (COPD). We used the Respiratory Quality of Life Questionnaire (RQLQ), a modification of an Australian questionnaire intended for asthma patients and adapted in this study to fit patients with COPD also. For comparison we chose the general health profile measure Short Form 36 (SF-36). We assessed the five RQLQ scales and eight SF-36 scales for reliability, validity and responsiveness in 59 outpatients attending a Norwegian hospital for COPD. Statistical analysis included internal consistency, test-retest reliability and convergent validity between the two questionnaires. Responsiveness was assessed in patients reporting global change in health status over 1 year. All scales of the RQLQ showed good internal consistency (Cronbach's a = 0.85-0.94) and test-retest reliability (intraclass correlation coefficient = 0.86-0.94), as did the SF-36 scales (a = 0.66-0.90) and intraclass correlation coefficient = 0.60-0.86). Pearson correlations between scales with similar items ranged from 0.54 to 0.76, supporting the construct validity of both questionnaires. The RQLQ had responsive scales, showing significant changes in the expected direction over 1 year. We conclude that the RQLQ showed an acceptable reliability, construct validity and responsiveness in COPD patients, encouraging further use of this questionnaire. PMID- 10464834 TI - Inhaled antibiotic therapy in non-cystic fibrosis patients with bronchiectasis and chronic bronchial infection by Pseudomonas aeruginosa. AB - The aim of this study was to investigate the long-term effectiveness and safety of inhaled antibiotic treatment in non-cystic fibrosis patients with bronchiectasis and chronic infection by Pseudomonas aeruginosa, after standard endovenous and oral therapy for long-term control of the infection had failed. After completing a 2-week endovenous antibiotic treatment to stabilize respiratory status, 17 patients were randomly allocated to a 12-month treatment either with inhaled ceftazidime and tobramycin (group A) or a symptomatic treatment (group B). One patient from group A abandoned inhaled treatment because of bronchospasm and another from group B died before the end of the study. The remaining 15 patients, seven from group A and eight from group B, completed the study. Both groups had similar previous characteristics. The number of admissions and days of admission (mean +/- SEM) of group A [0.6 (1.5) and 13.1 (34.8)] were lower than those of group B [2.5 (2.1) and 57.9 (41.8)] (P < 0.05). Forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV1), PAO2 and PACO2 were similar in the two groups at the end of follow-up, showing a comparable decline in these parameters. There were no significant differences either in the use of oral antibiotics or in the frequency of emergence of antibiotic-resistant bacteria between groups. Microbiological studies suggested that several patients had different Pseudomonas aeruginosa strains. None of the patients presented impaired renal or auditory function at the end of the study. This study suggests that long-term inhaled antibiotic therapy may be safe and lessen disease severity in non-cystic fibrosis patients with bronchiectasis and chronic bronchial infection by Pseudomonas aeruginosa which do not respond satisfactorily to antibiotics administered via other routes. PMID- 10464835 TI - Chronic obstructive pulmonary disease, with and without alpha-1-antitrypsin deficiency: management practices in the U.K. AB - Alpha-1-antitrypsin deficiency is a common genetic defect associated with the development of severe and rapidly progressive lung disease. This study was undertaken to determine whether respiratory physicians manage patients with alpha 1-antitrypsin (AAT) deficiency differently from patients with chronic obstructive pulmonary disease (COPD) without alpha-1-antitrypsin deficiency. In addition we obtained physicians' views on who should be tested for AAT deficiency. A questionnaire was administered to 88 respiratory physicians based throughout the U.K. (44 in teaching hospitals). The main outcome measures were pulmonary function tests, radiological assessment, frequency of repeat testing, follow-up and screening protocol for alpha-1-antitrypsin deficiency. Subjects with homozygous (PiZ) AAT deficiency were more likely to: 1. have baseline and full pulmonary function testing including dynamic flow rates, static lung volumes, and gas transfer; 2. have more comprehensive assessment with high resolution computed tomography (HRCT) thorax and repeated radiological assessment (with annual chest radiography); 3. be followed-up routinely; and 4. have family members tested for alpha-1-antitrypsin deficiency. Testing remains limited for AAT deficiency and is mainly restricted to young patients with COPD. COPD assessment and management is influenced by the presence of AAT deficiency, which may reflect the poorer prognosis of such patients due to more rapid decline. Assessment and monitoring could be simplified to forced expired manoeuvres, although limited HRCT thorax and tests of gas transfer may prove more sensitive to progression of emphysema. Testing for AAT deficiency in the U.K. remains restricted, which will influence the detection rate for AAT deficiency. A wider policy of testing was advocated by the WHO will detect more patients and also influence our understanding of the natural history of the condition. PMID- 10464836 TI - Cutaneous delayed-type hypersensitivity reactions in smokers with chronic bronchitis and recurrent exacerbations: comparison with asymptomatic smokers and never-smokers. AB - The aim of the present study was to investigate whether smoking patients with chronic bronchitis (CB) and recurrent exacerbations show signs of depressed cell mediated immunity (CMI), as reflected in the cutaneous delayed-type hypersensitivity (DTH) reaction, in comparison with asymptomatic smokers and healthy never-smokers. The study was a comparative clinical study performed at a university hospital center of respiratory medicine. Sixteen smokers with stable CB and recurrent exacerbations, five of whom had mild airflow obstruction, 18 asymptomatic smokers and 18 healthy never-smokers, all aged between 35 and 64 years, participated. No subjects treated with corticosteroids or N-acetylcysteine were included. Cutaneous DTH-reactions to seven recall antigens were assessed with Multitest, a standardized in vivo test of clinical CMI. Reactions were assessed 48 h after application by measurement of skin induration. A score (sum in mm of positive reactions) was created to assess overall reactivity. Neither the score nor the number of positive reactions differed significantly between the three study groups. Men had a significantly higher reactivity than women (P < 0.05) irrespective of group affiliation. No influence of smoking status on DTH reactivity could be seen. In the CB group no correlation was found between DTH reactivity and number of exacerbations the past 2 years. Patients with chronic bronchitis and recurrent exacerbations did not differ from asymptomatic smokers or healthy never-smokers with respect to cutaneous DTH reactions. Depression of CMI, as measured in this study, does not seem to be a primary factor behind recurrent exacerbations in smokers with CB. PMID- 10464837 TI - Bronchoalveolar lavage cell profiles in Wegener's granulomatosis. AB - Pulmonary involvement due to Wegener's granulomatosis (WG) can present radiologically either as diffuse infiltrates or as nodular and linear opacities. Clinical experience suggest that these radiological patterns are associated with different bronchoalveolar lavage (BAL) cell profiles, but this has not been examined formally. We compared the BAL cell profile in eight WG patients with diffuse infiltrates on chest X-ray, indicative of highly active pneumonitis, with corresponding findings in 37 patients with nodular, linear and focal low attenuation infiltrates on high-resolution computed tomography (HRCT) which reflected low-grade, mainly interstitial disease. A control group was composed of 11 patients with pulmonary sarcoidosis. Diffuse infiltrates occurred in association with high systemic disease activity and featured a neutrophilic BAL profile in the presence of generally normal BAL lymphocytes. HRCT findings suggestive mainly of interstitial disease were associated with either a lymphocytic BAL cell profile or a normal cell pattern. Patients with a lymphocytic cell profile generally had a preferential elevation of CD4+ cells in the BAL in the presence of a normal CD4/CD8 ratio in the blood. This was a common feature of WG and pulmonary sarcoidosis. In conclusion, highly active pneumonitis and pulmonary disease of low or moderate activity in WG are associated with disparate BAL cell profiles. It remains to be examined whether the preferential elevation of CD4+ cells in the latter condition reflects a common pathogenetic role of this subset of cells in WG and pulmonary sarcoidosis. PMID- 10464838 TI - Elevated plasma brain natriuretic peptide levels in chronic respiratory failure with cor pulmonale. AB - Elevated plasma brain natriuretic peptide (BNP) levels have been described in patients with congestive heart failure and acute myocardial infarction. We measured plasma BNP levels in patients with chronic respiratory failure to evaluate the correlation between plasma BNP levels and pulmonary haemodynamics. Plasma BNP levels were measured in 28 patients with chronic respiratory failure accompanied by three underlying diseases [14 with chronic obstructive pulmonary disease (COPD), seven with sequelae of pulmonary tuberculosis (sequelae Tbc) and seven with diffuse panbronchiolitis (DPB)] by immunoradiometric assay methods (IRMA). Twenty-one of 28 patients had already received oxygen supplementation and 16 of 21 patients were treated as outpatients with home oxygen therapy. Plasma BNP levels were significantly elevated in patients with chronic respiratory failure complicated by cor pulmonale (81.5 +/- 13.1 pg ml-1) compared to patients without cor pulmonale (13.3 +/- 2.7 pg ml-1, P < 0.001). As controls, plasma BNP levels in 10 patients with primary lung cancer were studied, and the results (3.5 +/- 1.0 pg ml-1) were not significantly different from those of patients with chronic respiratory failure without cor pulmonale. Plasma BNP levels in 12 healthy subjects were also studied, and the results (7.2 +/- 1.0 pg ml-1) were not significantly different from those of the control subjects. Plasma BNP levels showed a weak linear correlation with systolic pulmonary arterial blood pressure, estimated by Doppler echocardiography (r = 0.43; P = 0.068), but there was no significant correlation between BNP levels and the degree of hypoxaemia (r = 0.30; P = 0.138). Plasma atrial natriuretic peptide (ANP) levels in patients with chronic respiratory failure were also measured using the same samples. Plasma ANP levels were also significantly elevated in patients with chronic respiratory failure complicated by cor pulmonale (80.8 +/- 12.1 pg ml-1) compared to patients without cor pulmonale (26.1 +/- 4.4 pg ml-1, P = 0.003). A significant correlation was found between plasma BNP and ANP levels (r = 0.68; P < 0.001). Our results suggest that the plasma BNP or ANP level may be a useful indicator for detecting the presence of cor pulmonale in patients with chronic respiratory failure. PMID- 10464839 TI - Associations between an asthma morbidity index and ideas of fright and bother in a community population. AB - There is a need for simple asthma outcome measures for primary care which are not only valid in terms of their relationship with lung function but also in terms of pragmatic psychological constructs. This study assesses the usefulness of adding items on the degree of 'bother' and 'fright' caused by the condition to a previously validated simple asthma morbidity index. A postal questionnaire survey comprising a simple asthma morbidity index and questions on 'fright' and 'bother' was conducted in one general practice in the north-east of England. Responses were obtained from 570 individuals. Of these, 184 (32%) reported low, 133 (23%) medium and 253 (44%) high morbidity. Twenty-nine per cent of respondents had felt frightened by their asthma in the previous 4 weeks. Both the 'fright' and 'bother' items were significantly associated with the morbidity index. The addition of 'bother' and/or 'fright' questions may improve both the content, construct and predictive validity of the morbidity index, but this needs to be established prospectively. PMID- 10464840 TI - Pituitary reactivity, androgens and catecholamines in obstructive sleep apnoea. Effects of continuous positive airway pressure treatment (CPAP). AB - We studied the effects of chronic nocturnal hypoxaemia due to obstructive sleep apnoea syndrome (OSAS) on the hypothalamic-pituitary-thyroid and hypothalamic pituitary-testicular axes and on catecholamine and cortisol secretion. We investigated whether hormones other than catecholamines may serve as markers for chronic hypoxic stress and the possible effects of nasal continuous positive airway pressure (nCPAP) treatment on endocrine status. Nocturnal oximetry was performed in 16 male patients with OSAS diagnosed by polysomnography, immediately before nCPAP treatment and in 11 of the patients the oximetry was repeated after 7 months of nCPAP therapy. Plasma and urinary catecholamines, luteinizing hormone (LH) testosterone, cortisol, thyroid stimulating hormone (TSH), prolactin (PRL), and the response of TSH and PRL to a thyroid releasing hormone (TRH) challenge test were measured immediately before and after 7 months of nCPAP treatment. Subnormal LH and TSH and elevated serum cortisol as well as increased nocturnal urinary norepinephrine levels were found in patients prior to treatment; otherwise endocrine values were normal. There was a significant correlation between low pretreatment nocturnal arterial oxygen saturation and high plasma and urinary norepinephrine levels. The nCPAP treatment caused significant reduction in serum prolactin and TSH, and significant reduction in plasma epinephrine and urinary norepinephrine. The reduction in serum TSH and urinary norepinephrine was most pronounced in the subjects with the worst pretreatment nocturnal hypoxaemia. No other significant changes were found in basal hormone levels. The response to TRH challenge was normal before and after treatment and was not influenced by CPAP therapy. OSAS is associated with elevated catecholamine and cortisol and decreased TSH and LH levels but a normal response to TRH challenge and a normal androgen status. Apart from catecholamines, none of the hormones studied is likely to serve as a specific marker for chronic hypoxic stress. PMID- 10464841 TI - Acute adult asthma--assessment of severity and management and comparison with British Thoracic Society Guidelines. AB - To investigate the accuracy of clinical severity assessment of asthmatics and to compare emergency and subsequent ward management with British Thoracic Society (BTS) Guidelines, the records of all patients admitted for severe asthma (46) over a 5-month period to a District General Hospital were inspected. Variations from recommended management were revealed. Appropriate oxygen administration was often not provided in casually and patients frequently left hospital before their discharge criteria were attained: recommended diurnal variations in peak flow were exceeded in 26%. Eleven per cent of discharges were against medical advice, making provision of adequate management logistically difficult. Adherence to BTS guidelines on the need for arterial blood gas (ABG) analysis would have led to a failure to detect significant hypoxaemia in 25% of cases. This study identified substantial variations from BTS management guidelines. It is suggested that oximetry is necessary on arrival to guide selection for arterial blood gas analysis. PMID- 10464842 TI - Successive follow-up of chest computed tomography in patients with Mycobacterium avium-intracellulare complex. AB - The aim of this study was to evaluate the changes in chest CT findings examined successively in patients with Mycobacterium avium-intracellulare complex (MAC) infection. We carried out a retrospective study of 25 patients with MAC infection who had serial CT scans. Patients included 18 women and seven men with a median age of 66 years. Mean (+/- SE) follow-up interval between the first and second CT was 27.5 +/- 4.2 months. The serial chest CT scans were reviewed with consensus reading by two observers. At the first chest CT examination, we found the following: bronchiectasis (in 133 of 250 fields), cavity formation (11 of 250 fields), centrilobular nodules (167 of 250 fields), air-space disease (30 of 250 fields) and nodules (81 of 250 fields). The middle lobe and lingula were frequently involved. Centrilobular nodule scores improved in seven patients; disease progressed in nine patients and was stable in nine patients. In addition, bronchiectasis scores improved in four patients; disease progressed in 15 patients and was stable in six patients. The score of bronchiectasis in the second CT was significantly higher than in the first CT. In conclusion, our data suggest that centrilobular nodules and bronchiectasis are frequent observations in patients with MAC. In addition, progression of bronchiectasis appeared to be caused by MAC infection. PMID- 10464844 TI - A heated humidifier reduces upper airway dryness during continuous positive airway pressure therapy. AB - Upper airway dryness is a frequent side-effect of nasal continuous positive airway pressure (nCPAP) therapy in obstructive sleep apnoea (OSA). In this situation, heated humidification is often used. Alternatively, oily nose drops are frequently applied to relieve dryness. The present study aimed to investigate the efficacy of a heated humidifier in comparison with oily nose drops. Twenty four OSA patients complaining of serious nCPAP-related upper airway dryness were randomized to 6 weeks of treatment either with heated humidification (HC 100, Fischer & Paykel, Inc., Auckland, New Zealand) or oily nose drops (Colda-Stop, Desitin, Inc., Germany). The patients completed questionnaires on the degree and frequency of upper airway dryness, compliance with nCPAP, intention to terminate nCPAP and comfort during the nCPAP therapy. All 12 patients treated with heated humidification improved in terms of the degree and frequency of upper airway dryness, and reported greater comfort when using the nCPAP device. All patients in the heated humidification group intending to terminate nCPAP therapy because of upper airway dryness persisted with nCPAP on addition of humidification. In contrast, only five out of 12 patients (42%) in the oily nose drops group reported their degree of upper airway dryness to be improved (P = 0.003), only three patients (25%) reported an improvement in the frequency of upper airway dryness (P < 0.001), and only five patients (42%) reported greater comfort when using the nCPAP device with oily nose drops (P < 0.001). In the group using oily nose drops none of the three patients who intended to terminate nCPAP therapy persisted with nCPAP. Heated humidification is highly effective and superior to oily nose drops in reducing the symptoms of upper airway dryness during nCPAP. PMID- 10464843 TI - Prevalence of asthma and 'probable' asthma in the Asian population in Blackburn, U.K. AB - Local research had shown increased asthma admission rates in the Asian ethnic group in Blackburn, U.K. Patients also reported that asthma seemed to develop some years after first arrival in the U.K. A community prevalence survey of respiratory symptoms and asthma was undertaken in three practices with no special asthma interest. The questionnaire was administered by a Health Visitor and language link worker. Of the Asian patients in the practices, 96.6% were studied. The age distribution was similar to that of the local 1991 census. Of the patients, 181/1783 (10.2%) had diagnosed asthma but positive responses to individual questions suggested underdiagnosis of asthma. Asthma prevalence was higher in males up to age 20 (14.6% vs. 8.2%), and aged over 50 (16.5% vs. 10.5%), but higher in females aged 20-49 (5.6% vs. 9.2%). There were no correlations with social class or Jarman index, and no effect of country of origin or duration in the U.K. by multivariate analysis. The prevalence of diagnosed asthma at ages 5-9 and 10-14 was higher than in previous studies. Diagnosed asthma prevalence rates fell in the 20-49 age band but rose again in the over-50s. In all age groups the prevalence of asthma is probably underestimated. Asthma prevalence was not related to social factors. The data show that those born in the U.K. are more likely to describe regular symptoms and to be on regular treatment, but that for those born abroad there was an increasing rate of symptoms and medication use with increasing duration in the country. These observations confirm patient views but are explained by the age/sex distribution of those born in the U.K. compared to immigrants. PMID- 10464845 TI - Acute safety of beclomethasone dipropionate in a new CFC-free propellant system in asthmatic patients. AB - Hydrofluoroalkane-134a (HFA-134a) is a new chlorofluorocarbon (CFC)-free propellant for use in metered dose inhalers. It provides a more environmentally friendly alternative to CFC propellants because it does not contain chlorine which is responsible for ozone depletion by CFCs. Beclomethasone dipropionate (BDP) is widely used for inhalation asthma therapy and is most commonly delivered by a CFC propellant system. The present study evaluated the acute safety of BDP formulated with the new propellant (HFA-134a BDP) compared with BDP in a CFC 11/12 formulation by measuring the acute bronchial response in asthmatic patients. The study was conducted as a randomized, single-blind, placebo controlled, four-period cross-over trial. Asthmatic patients received eight inhalations of four treatment regimens (HFA-134a BDP, 1600 mg total dose; CFC 11/12 BDP, 2000 mg total dose; HFA-134a placebo and CFC-11/12 placebo) in random order over four study days. Forced expired volume in 1 s (FEV1) was measured before and 2, 10, 20, 40 and 60 min after inhalation of the study treatments. The number of coughs was counted from the start of the first inhalation to 60 s after the last inhalation. There were no statistically significant differences between the treatment groups for changes in FEV1, for the number of coughs or for the occurrence or severity of bronchoconstriction. In asthmatic patients withholding bronchodilators, the new HFA-134a BDP propellant system proved as safe and was as well tolerated as the current CFC-11/12 BDP system. The two propellant systems without active drug were also equally well tolerated. PMID- 10464846 TI - A comparison of two long-acting beta-agonists, oral bambuterol and inhaled salmeterol, in the treatment of moderate to severe asthmatic patients with nocturnal symptoms. The French Bambuterol Study Group. AB - This multicentre study was set up to compare the efficacies of two long-acting beta 2-agonists, oral bambuterol (20 mg nocte) and inhaled salmeterol (50 micrograms b.i.d.), for the treatment of moderate to severe asthmatics who were considered to be on optimal steroid/bronchodilator therapy, but continued to have troublesome nocturnal symptoms. The study was of double-blind, parallel-group design and comprised a 2-week run-in on previous maintenance therapy followed by a 6-week study treatment period. There were 117 randomized asthmatic patients aged 20-70 years (65 women and 52 men with a mean age of 45 and predicted FEV1 of 64%), who had been taking 800-2000 micrograms inhaled steroid and/or up to 20 mg oral steroid per day for at least 4 weeks. They were asked to complete daily diary cards, recording morning and evening PEF, daily symptoms, nocturnal awakenings, rescue medication and subjective tremor. There was a significant increase in both morning and evening PEF respectively, on bambuterol (28 l min-1, 20 l min-1, P < 0.05) and salmeterol (29 l min-1, P < 0.001; 23 l min-1, P < 0.01) when compared with run-in. The mean percentage fall in overnight PEF was reduced by 8.3% (P < 0.001) on bambuterol and by 6.8% (P < 0.001) on salmeterol. Nocturnal awakenings and daytime symptoms due to asthma were significantly lowered by both treatments, as was the consumption of rescue bronchodilator. Tremor scores were very low during both run-in and study treatments. No significant treatment difference between bambuterol and salmeterol was detected for any of the above variables. Once-daily oral bambuterol provides a highly effective alternative to twice-daily inhaled salmeterol for relief of nocturnal symptoms in patients with moderate to severe asthma. PMID- 10464847 TI - Dyspnoea is associated with pulmonary function impairment in exposed workers. AB - This study aimed to evaluate the prevalence of dysponea and its predictors in studies on several working male groups in British Columbia (BC), Canada (cedar sawmill, grain elevator, pulpmill, and aluminum smelter workers), and Tuscany (T), Italy, (shoe and furniture makers, millers, bakers, and pharmaceutical workers). We performed cross-sectional health studies (interviews and pulmonary function tests) for 2498 BC and 1474 T workers exposed to air contaminants, and 1110 BC and 243 T controls. Similar questionnaires and the same definitions were used in BC and in T. Pulmonary function tests were also performed. The participation rates were >92% in BC workers and 82% in T workers. The overall prevalence of moderate dyspnoea was not different in exposed BC and T workers in comparison with controls. Slight dyspnoea was significantly more frequent in BC workers, but not in T workers, with respect to controls. After adjusting for age, body mass index (BMI), smoking, current asthma, and chronic bronchitis, forced expiratory volume in 1 s (FEV1) and forced vital capacity (FVC) were found to be significantly associated with slight and moderate dyspnoea in BC workers, and slight dyspnoea in T workers. Isolated dyspnoea is associated with reduction in FEV1 and FVC in working populations, after adjusting for potentially confounding variables. PMID- 10464848 TI - Prescription and usage of long-term oxygen therapy in patients with chronic obstructive pulmonary disease in The Netherlands. AB - Long-term oxygen therapy (LTOT) has been shown to improve survival in hypoxaemic patients with chronic obstructive pulmonary disease (COPD). This has resulted in recommending the prescription of oxygen for at least 15 h day-1 in most European countries. In order to examine the prescription and usage of LTOT and to assess the adherence to international recommendations for its prescription, a survey was set up in a random sample of clients of the largest oxygen company in the Netherlands. After patients had been visited for an interview, additional postal surveys were sent to the physician who had prescribed LTOT and to the oxygen company. For 175 COPD patients the mean oxygen prescription and mean oxygen usage were 15.6 +/- 5.8 and 14.1 +/- 6.8 h day-1, respectively. In 62 patients (35%) oxygen was prescribed < 15 h day-1, more often by non-chest physicians than by chest physicians (P < 0.0001), and 91 patients (52%) used oxygen < 15 h day-1. Of 113 patients with a prescription > or = 15 h day-1, 39 (35%) used oxygen < 15 h day-1 and 74 for > or = 15 h day-1. The latter were prescribed oxygen for more h day-1, had been longer on LTOT, had a higher resting flow rate, were prescribed a concentrator, employed portable cylinders and used oxygen in public significantly more often than the former. We conclude that in a selected group of LTOT patients with COPD both oxygen prescription and usage were often inadequate, particularly if LTOT was prescribed by non-chest physicians. PMID- 10464849 TI - The differential effect of pentoxifylline on cytokine production by alveolar macrophages and its clinical implications. AB - The aim of this study was to investigate the effects of pentoxifylline (PTX) on the production of TNF-alpha, IL-1 beta, IL-6 and GM-CSF by lipopolysaccharide (LPS)-stimulated alveolar macrophages (AM). AM and peripheral blood monocytes (PBM) from 10 patients were cultured for 24 h in the presence of LPS (10 micrograms ml-1) and PTX at concentrations of 2.0 mM, 1.0 mM, 0.5 mM, 0.1 mM and 0.01 mM. TNF-alpha and GM-CSF were measured from the culture supernatants of both the AM and PBM from all 10 patients and IL-1 beta and IL-6 from the culture supernatants of the AM from five patients. The TNF-alpha production by AM was significantly suppressed in the presence of PTX at concentrations of 2.0 and 1.0 mM, while production of IL-1 beta, IL-6 and GM-CSF remained unaffected. In PBM cultures, PTX significantly suppressed the production of TNF-alpha and GM-CSF, at all tested concentrations. The present study provides evidence that PTX selectively suppresses the production of TNF-alpha by LPS-stimulated AM and may have a role in the treatment of lung diseases where TNF-alpha is involved. The mode of administration of PTX should take into account the suppressive effect of this drug on GM-CSF production by PBM. PMID- 10464850 TI - Cough induced by low pH. AB - Citric acid has been used as a tussive agent in the investigation of the cough reflex. The mechanism of cough stimulation remains unclear. We conducted studies to compare the cough response of citric acid to an organic (acetic acid) and an inorganic acid (phosphoric acid). We also compared the cough response of citric acid to capsaicin, a non-acid tussive stimulus. In study 1, 26 subjects inhaled equimolar concentrations of citric and acetic acid. In study, 2, 22 subjects inhaled citric acid and phosphoric acid in concentrations of similar pH. Eighteen subjects from study 2 completed a capsaicin cough challenge test. The cough response was recorded and the concentration causing two coughs, the cough threshold, calculated (D2). The correlation of cough thresholds between both citric and acetic acid (r = 0.79, 95% CI 0.37-0.90, P < 0.0001) and citric acid and phosphoric acid (r = 0.68, 95% CI 0.37-0.86, P = 0.0005) were significant. There was no correlation between the citric acid and capsaicin cough thresholds. These results show that an individual's cough response to acetic and phosphoric acid are similar to that caused by citric acid. Thus these acids may cause cough by a common mechanism such as disturbance of the pH of the airway surface liquid. Capsaicin does not appear to share this common pathway. PMID- 10464851 TI - Effects of inhaled nitric oxide and aerosolized iloprost in pulmonary veno occlusive disease. PMID- 10464852 TI - Pulmonary toxicity of chemotherapy and G/GM-CSF: a report of five cases. PMID- 10464853 TI - Work-related asthma in an aircraft engine mechanic. PMID- 10464854 TI - Treadmill exercise duration and dyspnea recovery time in chronic obstructive pulmonary disease: effects of oxygen breathing and repeated testing. PMID- 10464855 TI - Of Bourne, badgers, and a bovine 'TB alert'. PMID- 10464856 TI - Acute effects of nebulised epoprostenol in pulmonary hypertension due to systemic sclerosis. AB - Pulmonary hypertension often has a lethal outcome in systemic sclerosis and the treatment is challenging. Epoprostenol is a potent pulmonary vasodilator and its efficacy has been demonstrated when delivered by the intravenous and aerosolized routes. We report the haemodynamic and functional benefits of epoprostenol administered by inhalation to a spontaneously breathing patient with partially reversible pulmonary hypertension due to systemic sclerosis. Aerosolized epoprostenol, equivalent to the maximum tolerated intravenous dose (31.2 micrograms), produced a 58% fall in pulmonary vascular resistance, increased the cardiac output by 42% and improved functional performance by one MET (3.5 ml kg-1 min-1 of oxygen uptake) without any significant side-effects. Selective distribution of epoprostenol by the inhaled route may offer a new strategy for treatment of pulmonary hypertension. PMID- 10464857 TI - Sleep quality, carbon dioxide responsiveness and hypoxaemic patterns in nocturnal hypoxaemia due to chronic obstructive pulmonary disease (COPD) without daytime hypoxaemia. AB - In order to clarify whether nocturnal hypoxaemia (arterial oxygen saturation, SaO2 < 90%) may exist in the long-term before daytime hypoxaemia (PaO2 < 8.0 kPa) occurs in chronic obstructive pulmonary disease (COPD), 21 patients with stable severe COPD without daytime hypoxaemia (PaO2 > or = 8.0 kPa) were studied prospectively. Subjects were monitored twice by polysomnography (PSG) 12 months apart. Spirometry was performed, and diffusion capacity (DLCO) and hypercapnic respiratory drive response delta PI0.1 delta PCO2(-1)) were measured during the daytime in conjunction with polysomnography. At the start of the study our subjects had FEV1 %P (FEV1 as a percentage of predicted value) of 26.1 +/- 7.2%, a mean nocturnal nadir SaO2 of 83 +/- 5%, and a mean SaO2 during nocturnal hypoxaemic episodes of 88.0 +/- 0.7%. The patients' delta PI0.1 delta PCO2(-1) was 1.8 +/- 1.4 cm H2O kPa-1 (within the normal range). For the entire study group, no significant change in any lung function or blood gas parameter was noted during the year of observation, and nocturnal SaO2 remained unaltered. Stage I sleep decreased (P < 0.05) after 12 months. Prolonged stage I sleep was associated with nocturnal hypoxaemia at the second PSG. Five subjects developed daytime hypoxaemia and they showed poorer lung function but similar nocturnal hypoxaemia and delta PI0.1 delta PCO2(-1) level compared to the rest of the patients. Patients with sudden SaO2 dips had more pronounced nocturnal hypoxaemia and prolonged wakefulness than 'non-dippers'. In conclusion, the mean level of nocturnal hypoxaemia may persist unaltered for at least 1 yr. COPD patients with exclusively nocturnal hypoxaemia have a hypercapnic drive response within the normal range. Prolonged nocturnal hypoxaemia and reduced whole night oxygenation are associated with increased superficial sleep. Sleep fragmentation and high carbon dioxide sensitivity may be important defence mechanisms against sleep related hypoxaemia. The appearance of daytime hypoxaemia is preceded by a substantial deterioration in lung function, but by only a minor deterioration of nocturnal hypoxaemia. PMID- 10464858 TI - Health beliefs and compliance with inhaled corticosteroids by asthmatic patients in primary care practices. AB - The aim of this study was to determine factors associated with regular use of inhaled corticosteroids (ICS) by asthmatic patients in primary care practices. A cross-sectional survey was carried out over 12 family practices in the Philadelphia greater Metropolitan area. A total of 394 patients aged 18-49 years, who received medical care for asthma from their primary care physician and had been prescribed ICS between 1 January 1995 and 31 December 1996, were included. The study measured self-reported demographics, experience with asthma, use of and attitudes about ICS, and health beliefs in six domains. Only 38% of patients reported using ICS at least twice a day almost every day. The most frequently cited reasons for inconsistent or non-use of ICS were related to a belief that ICS were unnecessary during asymptomatic periods and to a general concern about side-effects. By logistic regression, factors associated with regular use of ICS were two patient health beliefs, namely the health belief of 'Active' participation in clinical decision-making with their physician (OR = 4.6, 95% CI 2.8, 7.5), and the health belief that asthma was a 'Serious' health problem (OR = 2.3, 95% CI 1.4, 3.7), and hospitalization for asthma within the previous 12 months (OR = 2.3, 95% CI 1.6, 4.6). Patients were more likely to report regular use of ICS if they saw themselves as active participants in their treatment planning and conceptualized asthma as a potentially serious illness. These results support the themes of patient education and shared decision-making between patients and physicians that are promoted by the Asthma Guidelines from the National Heart, Lung and Blood Institute (NHLBI). PMID- 10464859 TI - Clinical and laboratory features of lupus patients with complicating pulmonary disease. AB - The objective of this study was to determine the incidence of pulmonary involvement in patients with systemic lupus erythematosus (SLE) and to clarify the clinical and laboratory characteristics in SLE patients with various pulmonary involvements. A retrospective study (n = 137) revealed that the types of pulmonary involvement found in SLE patients were: pleuritis (9%), interstitial pneumonia (8%), pulmonary infarction (7%), pulmonary infection (4%), pulmonary hypertension (2%), restrictive dysfunction (28%) and decreased diffusion capacity (43%). The incidences of pericarditis (P < 0.01), arthralgia (P < 0.05) and hypoalbuminemia (P < 0.05) were significantly greater in patients with pleuritis than in those without, while in patients with interstitial pneumonia, the incidence of anti-SS-A antibody (P < 0.05) and sicca syndrome (P < 0.05) were significantly greater than in those without. A longitudinal follow-up study of patient groups with various pulmonary involvements revealed: 1. significant changes of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), lactate dehydrogenase (LDH) and thrombomodulin (TM) in patients with pleuritis, and 2. significant changes of WBC and LDH in patients with interstitial pneumonia. The increased ESR, CRP and TM levels during disease episodes suggest that the involvement of inflammatory processes is related to vasculitic events in the pathogenesis of lupus pleuritis. A higher incidence of anti-SS-A antibody in lupus patients with interstitial pneumonia suggests a potential role for this autoantibody in the pathogenesis of this complication. PMID- 10464860 TI - Agreement between spirometry and tracheal auscultation in assessing bronchial responsiveness in asthmatic children. AB - We have recently found that changes in lung sounds correspond well with a 20% fall in the forced expiratory volume in 1 s (FEV1) after methacholine challenge in asthmatic children. Up to now, little was known about the agreement between a 20% fall in FEV1 and a change in lung sounds after repeated bronchial challenge. In this study we investigated the agreement between the total cumulative histamine dose causing a fall in FEV1 of 20% or more (PD20) and the detection of a change in lung sounds (PDlung sounds) after two bronchial challenges on different occasions in asthmatic children. Fifteen asthmatic children (nine boys), mean age 10.8 years (range 9-15), were studied. All performed two histamine challenge tests on 2 days, with a 24 h to 1 week interval. Lung sounds were recorded over the trachea for 1 min and stored on tape. Lung sounds were analysed directly and also scored from the tape-recording by a blinded second investigator. Wheeze, cough, and an increase in respiratory rate were assessed. The relationship between PD20 and PDlung sounds was calculated by Bland and Altman's measurement of agreement. Eleven children had a positive challenge test (PD20 < or = 16.0 mg ml-1) on both test days; four had a positive challenge on one test day. In 24 out of 26 positive challenges, wheeze, cough, prolonged expiration and/or increased respiratory rate were detected one dose-step before, or at the dose-step of histamine that induced a fall in FEV1 of 20% or more. In two challenges, PD20 was not detected by a change in lung sounds. In four out of four negative challenges (PD20 > 16.0 mg ml-1) no change in lung sounds could be detected. Good agreement between the logarithm of PD20 and the logarithm of PDlung sounds was found on both test days. The mean difference was 0.04 and the limits of agreement (d +/- 2 SD of the differences) were 0.04 +/- 0.41. A good agreement was found between the total cumulative histamine dose causing a fall in FEV1 of 20% or more and the detection of a change in lung sounds after two bronchial challenges on different occasions in asthmatic children. PMID- 10464861 TI - Snoring and myocardial infarction: a 4-year follow-up study. AB - The association between snoring and myocardial infarction was studied in 1453 people of both sexes aged 20-70 years. The study was carried out in a population of 92,364 residents and the subjects were recruited using the Electoral Census. A questionnaire was sent to all participants, asking about snoring and cardiovascular risk factors. Hospital records were checked for the next 4 years to establish how many of them developed myocardial infarction. At the beginning of the follow-up study 39 patients were diagnosed with ischaemic heart disease. Of the other 1414 participants, 571 (40.4%) were snorers and 843 (59.6%) non snorers. Twenty-one developed myocardial infarction in the snorer group and four in the non-snorer group. The snorer group presents an adjusted relative risk of myocardial infarction of 3.08 (95% CI 1.01-9.46) with respect to non-snorers. We conclude that snoring seems to be a potential risk factor for myocardial infarction. PMID- 10464862 TI - Clinical features of non-specific interstitial pneumonia. AB - The clinical features of 24 patients with non-specific interstitial pneumonia (NIP) were evaluated. The patients consisted of seven men and 17 women, with a median age of 60 years. In seven patients, the disease was idiopathic and eight had collagen vascular diseases. Cough, dyspnoea and fever were frequently observed. The time interval between the onset of symptoms and open lung biopsy was 3 months. Mild increases of IgG, CRP, and LDH were also frequently observed. The average per cent VC was 65.1 +/- 3.2% and the average PaO2 was 71.3 +/- 2.4 Torr. Bronchoalveolar lavage was done in 20 patients, and a moderate increase in lymphocytes (27.8 +/- 6.7%) with a low CD4/CD8 ratio (0.86 +/- 0.22) was observed. Multiple patchy infiltration or diffuse interstitial shadows, located predominantly in the lower fields of both lungs were the characteristic chest CT findings. Lung biopsies in this group were characterized by varying proportions of chronic interstitial inflammation and fibrosis which was temporarily uniform. Patients were given steroid pulse therapy or oral steroids. The results were mild to marked improvements in chest roentgenographic findings and lung functions. Four patients (16.7%) died because of respiratory failure caused by NIP. This is the first report to describe clinical features of NIP since the original report by Katzenstein and Fiorelli. PMID- 10464863 TI - No relation of tuberculin reactivity with quantitative analysis of peripheral blood lymphocyte subsets in haemodialysis patients. AB - Anergic response to tuberculin (PPD) is well known in haemodialysis patients (HDp). This cross-sectional controlled study was conducted to analyse the correlation of PPD response with demographical features, nutritional parameters and the distribution of peripheral blood lymphocyte (PBL) subtypes. In this study 29 HDp (17 men, 12 women; mean age 30.9 +/- 9.5 years) and 13 controls (eight men, five women; mean age 29.2 +/- 6.4 years) were included. The mean time spent on dialysis was 20.5 +/- 17.4 months. The mean PPD response was lower in HDp than controls (7.5 +/- 8 mm vs. 15 +/- 4 mm, P = 0.001). Fourteen patients (48%) were PPD (-) (eight men, six women; mean age 34.1 +/- 11.1 years) and 15 were PPD (+) (normergic) (nine men, six women; mean age 26.8 +/- 3.4 years). No difference was observed between PPD (-) and (+) groups for age, sex and time spent on dialysis. As nutritional parameters, body mass index, serum albumin, creatinine and cholesterol levels were measured and no differences were found between controls and the PPD (-) and (+) groups. Absolute lymphocyte counts were lower in HDP compared to controls (1290 +/- 296 vs. 1570 +/- 307 cells ml-1; P = 0.01). PBL subtype percentages and absolute counts (CD3, CD4, CD8, CD4/CD8, HLADR+CD3+, CD16+56+, CD19) were also similar between PPD(-) and (+) HDp. It was concluded that PPD response cannot be predicted by the distribution of PBL subtypes. The most probable cause of this observation is regulation of PPD reactivity by in situ immune cells whose composition is not reflected in the distribution of PBL. PMID- 10464864 TI - Assessing lung deposition of inhaled medications. Consensus statement from a workshop of the British Association for Lung Research, held at the Institute of Biology, London, U.K. on 17 April 1998. AB - In vitro measurements of aerosol fine particle fraction (FPF) using particle sizing apparatus (e.g. the twin impinger, multi-stage liquid impingers, cascade impactors) have a key role to play in the development of new pharmaceutical products and in quality control. However, use of in vitro methodology to attempt to predict lung deposition in vivo is of limited value due, in part, to the inability of current apparatus to mimic upper and lower airway anatomy satisfactorily. Estimates of FPF based on cut-off points ranging from 5-7 microns generally overestimate lung deposition as measured in vivo by gamma scintigraphy. We recommend that: 1. multistage apparatus (minimum five stages) be used to characterize particle size distribution adequately, over the range 0.5-5.0 microns; 2. where possible, measurements should be made at a range of rates and profiles of flow reflecting those likely to be generated using the inhalation device in clinical practice (including use by young and elderly patients with varying degrees of airflow obstruction); 3. encouragement should be given to the further development, standardization, and validation of apparatus with a 'throat' which more closely resembles the human oropharynx and larynx. Pharmacokinetic methods can give a good estimate of total, but not regional, lung deposition, with drugs which are either not absorbed via the gastrointestinal tract, or whose absorption can be blocked by co-administration of charcoal, thus avoiding confounding by absorption of drug substance deposited in the oropharynx and subsequently swallowed. Techniques which rely on evaluation of a timed fractional output of drug substance in the urine are susceptible to the inherent variability of rate of absorption across the respiratory epithelium. We recommend that consideration should be given to the further refinement and validation of PK methods which would more clearly identify the fractional dose deposited in the lung. Lung-imaging methodology, e.g. gamma scintigraphy, employing formulations radiolabelled with gamma-ray-emitting radionuclides such as 99mTc, can measure total lung deposition and oropharyngeal deposition, provided that the radiolabelling process is appropriately validated and suitable corrections are made for attenuation of gamma rays by body tissues. An estimate of regional lung deposition can be made by drawing 'regions of interest' on the scintigraphic image; the precision of this measure is limited by the two-dimensional (2-D) nature of most images which mean that there is an overlay of structures of interest (alveoli, small and large airways), which is most marked centrally. Three-dimensional (3-D) imaging techniques (e.g. single photon emission computed tomography, SPECT, and positron emission tomography, PET) have the potential to give more detailed data on regional lung deposition, but are currently more expensive, employ higher radiation doses, and are less well validated than 2-D (planar) imaging. We consider that, of the available imaging modalities, planar gamma scintigraphy represents current best practice for the assessment of lung deposition from inhaler devices where regional differences may be important. The methodology should be optimized by the adoption of generally accepted standards for radiolabelling, imaging, attenuation correction, and interpretation. It is important that deposition in all sites (device, oropharynx, lungs, stomach) should be quantified. Consideration should be given to refining the concept of regions of interest to coincide more closely with anatomical lung structures. Statistical methods to compare the size distributions of drug and radiolabel in validation experiments should be developed. In the longer term it is envisaged that three-dimensional imaging may play a more important part in evaluating lung deposition; an optimal three-dimensional anatomical model of lung zones of interest needs to be developed. PMID- 10464865 TI - Nostril widening improves arterial oxygenation: a role for nasal nitric oxide? PMID- 10464866 TI - Hamman-rich syndrome 'primed' by radiation? PMID- 10464868 TI - Corticosteroid treatment of asthma: now at the crossroads. PMID- 10464867 TI - Pulmonary alveolar proteinosis with lung squamous cell carcinoma. PMID- 10464869 TI - Re: Asthma in the elderly: underperceived, underdiagnosed, and undertreated; a community survey. PMID- 10464870 TI - Inhaled corticosteroids for adult asthma: impact of formulation and delivery device on relative pharmacokinetics, efficacy and safety. AB - Metered dose inhalers (MDIs) are the mainstay of inhaled steroid therapy for asthma. With the phasing out of traditional chlorofluorocarbon (CFC) propellants and their replacement with a new generation of CFC-free products, it is becoming clear that formulation and inhaler characteristics can markedly affect the drug delivery. It now seems necessary to compare inhalers not only on the basis of the properties of the steroid molecules but also to take into account the effect of propellants and other inhaler characteristics. The impact of formulation and delivery device on relative pharmacokinetics, therapeutic efficacy and tolerability is illustrated by a new preparation of beclomethasone dipropionate (BDP) in an inhaler containing hydrofluoroalkane (HFA) propellant, called Qvar (3M Health Care, U.K.). This drug preparation delivers the majority of particles (60%) in the fine particle range. This appears to be associated with improved lung deposition, a halving of dose requirements of BDP, but no evidence of clinically relevant adrenal suppression when used in therapeutic doses. Prescribers need to be aware of the impact of formulation on pharmacokinetics of inhaled steroids in order to offer the lowest effective dose and give clear instructions to patients who are changing to a CFC-free product. PMID- 10464871 TI - An observational study of inhaled corticosteroid withdrawal in stable chronic obstructive pulmonary disease. ISOLDE Study Group. AB - Withdrawal of inhaled corticosteroids is known to worsen disease control in bronchial asthma but similar data are lacking in chronic obstructive pulmonary disease (COPD). We hypothesized that clinical exacerbations requiring treatment would occur more often in patients whose inhaled corticosteroids were stopped than in other patients not treated with these agents. We studied 272 patients in mean age 65 (SD 0.8) years, mean FEV1 42.8 (SD 12.6)% predicted, entering the run in phase of the Inhaled Steroids in Obstructive Lung Disease (ISOLDE) trial. All had been clinically stable for at least 6 weeks and there were no differences in the degree of bronchodilator reversibility, baseline lung function or pack-years of smoking between the 160 patients receiving inhaled corticosteroids and those not so treated. Inhaled corticosteroids were withdrawn in the first week of the study and during the remaining 7 weeks of the study 38% of those previously treated with these drugs had an exacerbation compared to 6% of the chronically untreated group. Patients receiving inhaled corticosteroids reported a longer duration of symptoms but neither this or any other recorded variable predicted the risk of exacerbation. These data suggest that abrupt withdrawal of inhaled corticosteroids should be monitored carefully even in patients with apparently irreversible COPD. PMID- 10464872 TI - New formulation metered dose inhaler increases breath alcohol levels. AB - A new, chlorofluorocarbon-free metered dose inhaler containing salbutamol, with small amounts of ethanol as a co-solvent, has recently been released. We evaluated the effect on breath alcohol levels of two inhalations from this metered dose inhaler. In 10 volunteers, breath alcohol levels rose to 17.9 micrograms 100 ml-1 breath after an inhalation with a good technique, and to 35 micrograms 100 ml-1 breath (the current legal limit for driving in the UK) after a poor inhalation. Breath alcohol levels were reduced by the use of a spacer device, and in all cases fell to near zero after 2 min. In patients who have just used this type of metered dose inhaler, breath alcohol measurements should be defferred for at least 2 min. PMID- 10464873 TI - A national program for detection of alpha 1-antitrypsin deficiency in Italy. Gruppo I.D.A. AB - alpha 1-antitrypsin (AAT) deficiency is an inherited condition characterized by low serum levels of AAT and an increased risk of developing pulmonary emphysema. The disease occurs mainly in Caucasians, but Southern Europe, including Italy, is considered a low prevalence area. We developed a national program in Italy in order to improve our knowledge of the epidemiology of AAT deficiency and to establish a registry of the AAT-deficient individuals. The program had two phases: the first lasted 36 months, during which blood from coupons mailed by respiratory physicians from throughout the country, was isoelectrofocused by the Central Laboratory in Rome. The second phase started in February 1996, and the Registry was established. Up to August 1998, 151 subjects with AAT deficiency have been identified and 64 have been enrolled in the Registry. We believe that such a program plays a crucial role in identifying AAT deficiency in a country such as Italy, with low prevalence and low awareness of this rare condition. PMID- 10464874 TI - Treatment of chronic obstructive pulmonary disease and its exacerbations in general practice. EOLO Group. Estudio Observacional de la Limitacion Obstructiva al Flujo aEreo. AB - The high prevalence and chronicity of chronic obstructive pulmonary disease (COPD) imply that many of these patients are treated and controlled in primary care centres, often without contact with specialized pneumologist care. We conducted the present study to evaluate the treatment administered in stable and exacerbated COPD in GP-setting clinics and to investigate which factors could be associated with the different prescriptions. This is a cross-sectional observational study of ambulatory COPD patients. General practitioners (n = 201) were selected throughout Spain by regionally stratified sampling. We recorded the physician-reported prescription drug use in ambulatory treatment of stable COPD and acute exacerbations of COPD through a standard questionnaire. Factors independently associated with the prescription of drugs were ascertained by multiple logistic regression analysis. Of 1078 questionnaires reviewed, 1001 fulfilled quality criteria. There were 878 men (88%) and 123 women (12%); 777 (78%) were smokers or ex-smokers with a mean age of 68 years. Mean FEV1 was 47% predicted (% pred.) (SD = 13%). The median number of exacerbations was two per year (range = 0-16). Regular treatment for COPD was received by 878 (88%): the most commonly used drugs were inhaled beta 2-agonists (71%), theophyllines (53%) and inhaled corticosteroids (ICs) (50%), followed by mucolytics (25%), ipratropium bromide (23%), and oral corticosteroids (OCs) (4%). Treatment for exacerbations included inhaled bronchodilators (90%), antibiotics (89%), ICs (71%) and OCs (43%). Impairment of FEV1 was the factor most strongly associated in multiple regression analysis with increasing drug prescription in stable COPD, except for mucolytics, while the number of previous acute exacerbations was the main factor associated with exacerbation treatment except for OCs, the use of which was associated with more impaired pulmonary function. A significant number of the treatments prescribed in primary care for stable and exacerbated COPD do not follow current recommendations. Impairment in FEV1 is the factor most strongly associated with increasing prescription in stable COPD and the number of previous exacerbations is the main factor associated with exacerbation treatment. PMID- 10464875 TI - Effect of nebulized albuterol on circulating leukocyte counts in normal subjects. AB - Nebulized beta 2-receptor agonists may cause neutrophil demargination and result in misleading total circulating leukocyte counts (WBCs) in patients with acute bronchospasm. Varying underlying adrenergic stimulation in these patients also makes interpretation of these data difficult. This study examined the direct effect of these agents on the measured WBCs of healthy adults without evidence of bronchospasm or illness. A prospective, blinded, randomized study of 30 healthy volunteers (aged 18-50 years) was performed in a controlled environment. Subjects were excluded if they were pregnant, had a known underlying medical disorder or have had a prior reaction to albuterol or similar medications. Participants in the study were given either a nebulized albuterol treatment or nebulized normal saline (control group). Leukocyte counts were then obtained before and after treatments. Paired data were analysed using a one-tailed t-test while considering an increase of 40% in WBCs to be significant, P = 0.05, and beta = 0.10. Mean leukocyte counts were 5.9 (+/- 1.2) before treatment as compared to 6.0 (+/- 1.3) after albuterol nebulization. Using the coefficient of variance of WBCs in normal humans as c. 50% (6000 +/- 3000 cells mm-1) we were unable to demonstrate a significant difference in variation in post-nebulized leukocyte counts between the control group and the nebulized albuterol group. While there is concern that the treatment of patients experiencing acute bronchospasm with beta 2 agonists may result in factitious elevations in peripheral leukocyte counts, were found no direct effect of these agents on measured counts in normal subjects. PMID- 10464876 TI - Eosinophilic inflammation assessed by induced sputum in corticosteroid-dependent asthma. AB - Sputum induction can be used to study airway inflammation in asthmatics. However, it has not been used in patients with corticosteroid-dependent asthma requiring long-term oral corticosteroids. The aim of the study was to assess the number of eosinophils and the levels of eosinophil cationic protein (ECP) in sputum of 17 corticosteroid-dependent asthmatics by comparison with nine mild untreated asthmatics, 10 moderate asthmatics receiving inhaled steroids (ICS) and 11 healthy subjects. In the 17 corticosteroid-dependent asthmatics, we examined sputum eosinophil markers on two occasions and correlated with the control of asthma. Eosinophils were undetectable in controls and were detected in 63.8% of asthmatics. There were no significant differences between the three groups of asthmatics. ECP levels were significantly increased in ICS or corticosteroid dependent asthmatics by comparison to controls and mild asthmatics. There was no significant difference between ICS and corticosteroid-dependent asthmatics. During follow-up, corticosteroid-dependent asthmatics with a controlled disease had no significant change in eosinophil numbers or ECP levels. On the other hand, corticosteroid-dependent asthmatics with recent exacerbations had a non significant increase in eosinophil numbers and a significant increase in ECP levels. This study shows that ECP levels may be more accurate than eosinophil numbers in assessing exacerbations in corticosteroid-dependent asthmatics. PMID- 10464877 TI - The role of the expiratory phase in obstructive sleep apnoea. AB - The role of the expiratory phase in obstructive sleep apnoea (OSA) is not well known. The aim of our study was to verify the contribution of expiratory narrowing to apnoea in a group of OSA patients by evaluating the effects of short term treatment with continuous positive airway pressure (CPAP), intermittent positive pressure ventilation (IPPV) and bi-level positive airway pressure (BIPAP). We studied a selected group of 10 OSA patients whose therapeutic pressure level of CPAP was at least 10 cm H2O. During CPAP therapy the mean apnoea/hypopnoea index (AHI) and oxyhaemoglobin desaturation index (ODI) decreased from 64.8 to 6.3 (P < 0.001) and from 58.5 to 6.1 (P < 0.001), respectively and mean nadir SAO2 increased from 62.0 to 91.6 (P < 0.001). None of the patients reached optimal setting (elimination of snoring, reduction of apnoeas and non-apnoeic desaturation events at least to 15 or less per hour of sleep and maintenance of oxygen saturation approximately 90%) during IPPV and two patients did not tolerate final IPAP pressure levels. When a critical level of EPAP (BIPAP) was applied in the same night to these patients, optimal setting was reached in all subjects. During BIPAP, mean AHI decreased from 64.8 to 7.4 (P < 0.001); ODI decreased from 58.5 to 7.6 (P < 0.001) and nadir SAO2 increased from 62.0 to 91.2 (P < 0.001). Our study confirms the essential role of a critical level of EPAP in successful ventilatory treatment in OSA, thereby indicating, in agreement with few previous studies, the critical role of end of expiratory occlusion in OSA pathogenesis. PMID- 10464878 TI - The effect of thoracentesis on lung function and transthoracic electrical bioimpedance. AB - This study aimed to determine the relationship between improvement in lung function and changes in transthoracic electrical bioimpedance (TEB) after thoracentesis in patients with pleural effusions. Fifteen patients with pleural effusions due to either malignant (n = 8) or cardiac (n = 7) diseases were included. Pulmonary function was assessed before and after thoracentesis. During thoracentesis the patients were monitored with TEB. Using linear correlation analysis, the increases for each litre of aspirated thoracic fluid were: forced expiratory volume in 1 s (FEV1) 0.261; forced vital capacity (FVC) 0.331; total lung capacity (TLC) 0.58; and the lung diffusing capacity (DLCO); 2.4 ml min-1 mmHg-1. Baseline impedance increased by 2.3 Ohm l-1 aspirated thoracic fluid. The relative increase in baseline impedance was twice as high for patients with cancer as for patients with heart failure (P < 0.05). We found only minor changes in systolic blood pressure and mean arterial pressure. The improvements in diffusing capacity, airflow, and lung volumes after thoracentesis are correlated to an increase in baseline impedance, but changes are dependent on the primary disease. PMID- 10464879 TI - Predictors of nocturnal oxygen desaturation in patients with COPD. AB - The aim of this study was to identify factors which might predict nocturnal desaturation (defined as a fall of > 4% from awake baseline level for > or = 5 min) in normoxic or mildly hypoxic patients with stable COPD [arterial O2 saturation (SaO2) > or = 91%]. The study was prospective in nature, had full ethical approval and was performed in the Respiratory Department of a city teaching hospital. Thirty-three patients [mean (SD) age 67.2 (9) years] with stable COPD [mean (SD) FEV1 36.8 (11.0)% pred.] were recruited via the respiratory outpatient clinics and through the respiratory wards. The following parameters were measured: daytime arterial blood gases; spirometry; lung volumes (helium dilution); single breath CO transfer factor (TLCO and KCO); maximum inspiratory (IMP) and expiratory mouth pressures; pulse oximetry (SpO2) across a 6-min walk test, and SpO2 during sleep. Seventeen patients who experienced nocturnal desaturation had significantly lower mean PaO2 and SaO2, and higher PaCO2 values compared to non-desaturators. There was a positive correlation between mean nocturnal SpO2 and daytime PaO2, SaO2, and minimum exercise SpO2, and a negative correlation between mean nocturnal SpO2 and PaCO2, and FRC. Regression analysis revealed that daytime SaO2 was the only independent predictor of mean nocturnal saturation (accounting for 61% of the variability in the mean nocturnal SpO2). We observed nocturnal desaturation in all patients with a daytime SaO2 < or = 93% but in no patient with SaO2 > or = 95%. We conclude that daytime SaO2 can be used to predict nocturnal desaturation in normoxic or mildly hypoxic patients with stable COPD. Nocturnal desaturation is likely in patients with COPD where daytime SaO2 < or = 93%, and unlikely where daytime SaO2 > or = 95%. PMID- 10464880 TI - Utility of blood cultures in community-acquired pneumonia requiring hospitalization: influence of antibiotic treatment before admission. AB - It has been previously shown that antibiotics given before hospitalization significantly reduce the proportion of positive blood cultures in community acquired pneumonia (CAP). The aim of this prospective study was to compare the utility and cost-benefits of blood cultures in patients, hospitalized for moderate CAP, who had or had not received antibiotic therapy prior to admission. During 1 year, 53 patients were included and separated into two groups: group 1 patients had not received antibiotic treatment prior to admission (n = 30), whereas group 2 patients had been treated with antibiotics (n = 23). Within the first 48 hours, a set of blood cultures was collected if the body temperature was higher than 38.5 degrees C or in the case of shaking chills. A total of 136 blood cultures was collected; 74 in group 1 and 62 in group 2. Bacteraemia was significantly more frequent in group 1 than in group 2, 5/30 patients vs. 0/23, respectively (P < 0.05). The cost of negative blood cultures was valued at 13,939.2 FF in group 1 and 13,164.8 FF in group 2, respectively 464.6 +/- 244.3 FF and 569.3 +/- 233.4 FF per patient (n.s.). Moreover, blood cultures were the method of diagnosis in only one of the five patients with bacteraemia and in no case did a positive blood-culture result influence the initial therapeutic regime. Thus, our results suggest a reduced clinical utility and cost-benefit of blood cultures in patients hospitalized for moderate CAP who have received an antibiotic treatment prior to admission. PMID- 10464881 TI - Consumption of omega-3 and omega-6 fatty acids in former East and West Germany and changes in East Germany after the reunification. AB - The dietary intake of Omega-3 (omega-3) and Omega-6 (omega-6) fatty acids (FA) may influence the development of atopic diseases. Based on the results of two dietary surveys, the intake of omega-3-FA and omega-6-FA in West Germany in 1985 1989 and in Dresden, East Germany in 1988-1989 and 1990-1991 was estimated. The intake of omega-6-FA but not of omega-3-FA was slightly higher in the West German population before 1990, mainly because of a higher intake of vegetable oil. Immediately after 1990 the omega-6-FA-intake in the observed East German sample increased because of an increase in the consumption of margarine [from a mean of 16 (SD 17) to 38 (24) g day-1]. Additional information indicates that, in contrast, East German infant formulas contained higher amounts of omega-6-FA (linoleic acid) and lower amounts of omega-3-FA (alpha-linolenic acid) than West German formulas. Whether there is a causal relation between the amounts of omega 6-FA and omega-3-FA consumed and the prevalence of atopy in children from East and West Germany remains to be clarified. PMID- 10464882 TI - A thoracic complication of hereditary multiple exostoses in an adult. PMID- 10464883 TI - A critical reappraisal of transmission routes for bovine TB in cattle. PMID- 10464884 TI - Does an inhaled beta-adrenergic or anticholinergic agent improve gas exchange at rest and during exercise in patients with COPD? PMID- 10464885 TI - Should long-acting beta 2-agonists be considered an alternative first choice option for the treatment of stable COPD? PMID- 10464886 TI - Efficacy of once- and twice-daily administration of budesonide via Turbuhaler as initial therapy in patients with mild persistent asthma. AB - Inhaled glucocorticosteroids such as budesonide have an important role in the management of asthma. Although these agents have traditionally been given twice daily, evidence is accumulating that once-daily treatment may be as effective as twice-daily administration. The efficacy of budesonide Turbuhaler (Astra, Lund, Sweden), 400 micrograms once daily in the evening or 200 micrograms twice daily, was compared in a randomized, double-blind study involving 181 patients (75 men, 106 women; mean age 30.8 years) with mild asthma [mean forced expiratory volume in 1 s (FEV1) 92.8% pred.] who had not previously been treated with inhaled glucocorticosteroids. After a 2-week run-in period, patients were randomized to either regimen and treated for 6 weeks. This was followed by two 8-week open treatment periods, during which all patients received budesonide Turbuhaler, 200 micrograms once daily during the first period and 100 micrograms once daily during the second period. The mean change in morning peak expiratory flow (PEF) during the double-blind treatment period was 16.9 l min-1 in patients receiving once-daily treatment and 17.2 l min-1 in those receiving twice-daily treatment. Similarly, there were no significant differences in evening PEF, symptom scores, bronchodilator use or spirometry data between patients receiving once- and twice daily treatments. The improvements in morning PEF, symptom scores and bronchodilator use seen during the double-blind treatment period were maintained during the two open treatment periods. It is concluded that once-daily treatment with budesonide Turbuhaler is as effective as an initial therapy twice-daily treatment in patients with mild persistent asthma and that the initial dose can be reduced to maintenance levels (including 100 micrograms) without loss of asthma control. PMID- 10464887 TI - A comparison of the efficacy of long-acting beta 2-agonists: eformoterol via Turbohaler and salmeterol via pressurized metered dose inhaler or Accuhaler, in mild to moderate asthmatics. Force Research Group. AB - Four hundred and sixty nine patients were randomized to receive either 12 micrograms bd of eformoterol (Oxis, Astra Pharmaceuticals Ltd., Kings Langley, U.K.) delivered via Turbohaler or 50 micrograms bd salmeterol (Serevent, Glaxo Wellcome Ltd., Uxbridge, U.K.) via either the Accuhaler (Glaxo-Wellcome Ltd.) or pressurized metered dose inhaler (pMDI, Glaxo-Wellcome Ltd.) for 8 weeks. This was followed by a 4-week cross-over period when patients who had received salmeterol in the previous 8 weeks were given eformoterol and patients who had received eformoterol were given either salmeterol via the Accuhaler or pMDI to assess patient device and treatment preference. For the primary efficacy variable, the increase in peak expiratory flow (PEF) rate from run-in to 8 weeks, similar significant improvements were seen in all three treatment groups. Eformoterol Turbohaler (FT) achieved a greater increase in morning PEF than salmeterol Accuhaler (SA) from randomisation to 4 weeks; the increase shown in the eformoterol Turbohaler group was 28.9 l min-1 compared to 19.9 l min-1 for the salmeterol Accuhaler group. The addition of eformoterol Turbohaler 12 micrograms bd, to patients' existing asthma therapy was found to have a significantly more beneficial effect on the severity of patients' daytime asthma symptoms than had salmeterol Accuhaler 50 micrograms bd (P = 0.014). Eformoterol Turbohaler reduced the severity of daytime asthma symptoms by 42% after only 4 weeks of treatment. The patients in the eformoterol Turbohaler treated group experienced a higher percentage of days when they were symptom-free and did not use their short-acting bronchodilator to relieve symptoms (32.8, 24.1 and 28.0% in the FT, SA and SM groups, respectively). At 8 weeks there were no significant differences in any of these variables between the three groups. Patients in all the treatment groups gained an additional 1-1.5 nights undisturbed by asthma per week. The changes in sleep disturbance were not significantly different between the three treatment groups. In addition to the therapeutic benefits provided by eformoterol Turbohaler the device (Turbohaler) was the significant preference of patients given both Turbohaler and pMDI (P = 0.0168) and was also considered to be significantly more convenient to carry around than the Accuhaler (P < 0.0001). No other differences were found between the three devices. The results of this study demonstrate that the addition of a long-acting B2-agonist is an effective tool for achieving the goals of asthma treatment. Eformoterol via the Turbohaler is at least as effective as salmeterol via either the Accuhaler or the pMDI in achieving these goals. PMID- 10464888 TI - Inhaled beclomethasone (BDP) with non-CFC propellant (HFA 134a) is equivalent to BDP-CFC for the treatment of asthma. AB - As part of a development programme for a range of new CFC-free beclomethasone dipropionate (BDP) inhalers, two multicentre double-blind studies have been conducted to compare the therapeutic equivalence of a new HFA-134a propellant formulated BDP metered-dose inhaler (Norton Healthcare Ltd, London, U.K.) with a CFC counterpart for the management of adult patients with all grades of asthma. Doses of 100 micrograms qds for 6 weeks were administered in a low dose study and in a high dose study 500 micrograms qds doses were given for 12 weeks. Efficacy assessments included lung function (FEV1) in the clinic and asthma symptoms, peak flow rates and bronchodilator use by patients on diary cards. Safety parameters measured included routine haematology and biochemistry (including serum cortisols), clinical adverse events and throat swabs for Candida spp. Both CFC and HFA-formulations of inhaled BDP produced similar and significant improvements in lung function and asthma symptoms. In the low dose study, baseline to endpoint FEV1 increased from 2.2 +/- 0.51 to 2.5 +/- 0.81 (P = 0.0001) with BDP-CFC and from 2.2 +/- 0.51 to 2.6 +/- 0.81 with BDP-HFA (P = 0.0001), with no significant difference between treatments. In the high dose study, corresponding increases were 2.1 +/- 0.71 to 2.4 +/- 0.91 (P = 0.0002) for BDP-CFC and 2.1 +/- 0.71 to 2.3 +/- 0.71 (P = 0.017) for BDP-HFA. PEF also improved similarly on both treatments in both studies. Both formulations were well tolerated with no difference in the pattern of adverse events, effect on serum cortisol or Candida colonization. These studies showed that, in the management of asthma, the new HFA formulated BDP metered dose inhaler is equivalent to, and directly substitutable for, the older CFC-formulated product at the same dose, making change-over for patients straightforward. PMID- 10464889 TI - The efficacy and safety of a new ciprofloxacin suspension compared with co amoxiclav tablets in the treatment of acute exacerbations of chronic bronchitis. AB - A multinational, multicentre, randomized, prospective, parallel-group study compared treatment with ciprofloxacin administered as an oral suspension (500 mg twice daily for 7 days) with co-amoxiclav tablets (625 mg three times daily for 7 days) in patients suffering from acute exacerbations of chronic bronchitis (AECB). A total of 147 of 165 cases treated with ciprofloxacin (89.1%) and 146 of 162 cases treated with co-amoxiclav (90.1%) were classified as being clinical successes at the primary efficacy assessment 7 days after the end of therapy (assessed as reduced cough, improvement in dyspnoea, reduction in 24-h sputum volume or reduced purulence of sputum). Treatment equivalence was statistically confirmed; treatment difference:--1.0%, 95% CI--6.6% and 4.5%. Before treatment, 128 bacterial strains were isolated from 103 patients (60 ciprofloxacin and 68 co amoxiclav). The most commonly isolated organism was Haemophilus influenzae (60 isolates), followed by Moraxella catarrhalis (12 isolates), Streptococcus pneumoniae (11 isolates) and Staphylococcus aureus (10 isolates). At day 14, 40 of 46 ciprofloxacin-treated patients (87.0%) and 46 of 55 co-amoxiclav-treated patients (83.6%) who were valid for bacteriological analysis were classified as being bacteriological success (classed as eradication, eradication with colonization or presumed eradication; treatment difference: 3.3%, 95% CI--8.3% and 14.9%). The adverse event profile was comparable between treatment groups. Most adverse events considered possibly or probably related to study drug were related to the gastrointestinal system and were of mild or moderate severity: nausea (13% ciprofloxacin, 10.6% co-amoxiclav), flatulence (10.3% ciprofloxacin, 3.9% co-amoxiclav), abdominal pain (7.6% ciprofloxacin, 7.3% co-amoxiclav) and diarrhoea (4.3% ciprofloxacin, 6.7% co-amoxiclav). We concluded that a 7-day course of ciprofloxacin suspension is equivalent to a 7-day course of co amoxiclav tablets in terms of clinical and bacteriological efficacy and tolerability for the treatment of AECB. Thus, ciprofloxacin suspension may offer a suitable alternative treatment for AECB patients who have difficulty in swallowing, or who prefer liquid medications to tablets. PMID- 10464890 TI - Analysis of pleural fluid in idiopathic spontaneous pneumothorax; correlation of eosinophil percentage with the duration of air in the pleural space. AB - Pleural fluid analysis was performed in patients with idiopathic spontaneous pneumothorax. The objective of the study was to define the cell differentiation, and part of the cytokine profile, in relation to the duration of pneumothorax. In the 23 consecutive patients (19 men, mean age 34.2 years, 17 smokers), pleural fluid was obtained immediately after chest tube drainage (n = 6), or during thoracoscopy (n = 17). Cytospins were carried out, and supernatant analysis of the different cytokines was performed using sandwich ELISA. All concentrations were corrected for dilution. The duration of the pneumothorax was correlated with the rise in eosinophil percentage (r = 0.81, P < 0.00001) in pleural fluid. RANTES, platelet-activating factor (PAF), and monocyte chemotactic protein-1 (MCP 1) were detectable but no relationship with eosinophils or duration of the pneumothorax was found. Granulocyte-macrophage colony stimulating factor (GM-CSF) and interluekin-8 (IL-8) were not detectable. Interleukin-5 (IL-5) concentration correlated with the eosinophil concentration (r = 0.84, P = 0.037) and the eosinophil percentage (r = 0.68, P = 0.005) in the pleural fluid. Idiopathic spontaneous pneumothorax causes a time-related rise in the eosinophil percentage in the pleural space, which correlates with the level of IL-5. PMID- 10464891 TI - Serum leptin in children with asthma treated with inhaled budesonide. AB - Leptin, a 167-amino-acid peptide, is a recently discovered hormone which is believed to play a major role in the regulation of body weight. Systemic administration of exogenous glucocorticoids has been found to increase circulating leptin levels. In this study, we aimed to assess serum leptin in children with asthma treated with inhaled budesonide 800 micrograms day-1. Ten boys and three girls with asthma, all adolescents aged from 12.9 to 16.6 years, were studied in a randomized double-blind two-period cross-over trial with 4-week treatment periods and a 1-week wash out. Placebo was given during one period and 800 micrograms budesonide during the other via a 750 ml volume spacer (Nebuhaler, Astra Draco, Lund, Sweden). On the last day of the placebo and budesonide periods blood samples were taken and serum leptin was measured by a specific radioimmunoassay. The difference in mean (SEM) leptin concentration between the budesonide and placebo period was 0.2 (0.4) microgram l-1 (P = 0.71; t = -0.4; df = 12, 95% confidence interval -0.9-0.7 microgram l-1). Inhaled budesonide 800 micrograms per day from a Nebuhaler does not influence circulating leptin levels, suggesting that regulation of body weight is unaffected. PMID- 10464892 TI - Increased serum concentrations of conjugated diens and malondialdehyde in patients with pulmonary tuberculosis. AB - During pulmonary inflammation increased amounts of reactive oxygen species (ROS) and reactive nitrogen intermediates (RNI) are produced as a consequence of phagocyte respiratory burst. One of the manifestation of these free radical mediated processes is lipid peroxidation (LP). The aim of our study was to assess the concentration of lipid peroxidation products (LPPs), conjugated diens (CD) and malondialdehyde (MDA), in patients with active TB. Forty-two patients were enrolled into the study. Half (group I) had advanced TB and were sputum smear positive. The remainder (group II) had only small radiographical changes and were sputum smear-negative. Serum concentrations of CD and MDA were measured at days 0, 7, 14 and 28 in group I and day 0 in group II. We found that in all patients with active TB CCD (1.0 +/- 0.05A233) and CMDA (2.01 +/- 0.16 nmol dl-1) were significantly elevated compared to healthy controls (0.67 +/- 0.03A233 and 1.36 +/- 0.08 nmol dl-1, respectively) (P < 0.001). The highest levels of LPPs were in patients with advanced TB. These concentrations were stable during the first month of anti-tuberculous therapy. Our data indicated that, as in bacterial pneumonia, LPPs were enhanced in active TB. The levels of LPPs depended on the form of the disease as they were higher in subjects with advanced disease than in those with only small radiographical changes. Further studies are needed to assess the role of antioxidants as adjuvant therapy in patients with pulmonary TB. PMID- 10464893 TI - Methacholine-induced fall in forced vital capacity as a marker of asthma severity. AB - The dose of methacholine causing a 20% fall in forced expiratory volume in 1 sec (FEV1) from baseline (PD20) has been used as an index of asthma severity. The aim of this study was to determine if the percentage fall in forced vital capacity (FVC) from baseline at the PD20 (dFVC%) is an independent marker of asthma severity. We first retrospectively studied the dFVC% and PD20 obtained from 149 consecutive newly diagnosed asthmatics with a positive methacholine-challenge test (MCT). We then performed MCT on 20 normal subjects and 35 stable asthmatics. The 'milder' asthmatics (n = 20) and 'more severe' asthmatics (n = 15) were on regular inhaled corticosteroids: 200 micrograms or less and 800 micrograms or more daily, respectively. A dosimeter technique was used, and normal subjects were given a cumulative dose of 2400 micrograms. The PD20 and dFVC% were calculated using log-linear interpolation of the last two points. Student's unpaired t-tests and linear regression analyses were used for comparison and correlation of results. There was no significant correlation between dFVC% and PD20 among the 149 newly diagnosed asthmatics (r = 0.1), or among the 35 known stable asthmatics (r = 0.008). The more severe asthmatics had a larger dFVC% compared with the milder asthmatics (15.8% vs. 9.6%; P = 0.0005). In addition, inhaled corticosteroid usage correlated better with dFVC% (r = 0.56) than with PD20 (r = 0.36). The normal subjects had a mean fall in FVC of only 4.8%. The percentage fall in FVC at PD20 (dFVC%) may be a useful index of asthma severity which is independent of PD20. This index is potentially complementary to the PD20 in the assessment of asthma severity. PMID- 10464894 TI - Long-term (3-year) economic evaluation of intensive patient education for self management during the first year in new asthmatics. AB - Patient education and self-management programmes have proved effective in many studies with short follow-up periods. We studied the 3-year cost-effectiveness of an intensive programme of patient education and supervision for self-management. The study consisted of 162 consecutive newly diagnosed asthmatics who were randomized either into an intervention group (IG) receiving intensive patient education and supervision for self-management at an outpatient clinic during the first year, or a control group (CG) receiving conventional education at the baseline visits only. Both groups had 2 additional years of follow-up. Lung functions and health-related quality of life (HRQOL) were measured. Extra direct and indirect costs were recorded. At 3 years the differences in forced expiratory volume in 1 s (FEV1) and in peak expiratory flow (PEF) were significantly better in the IG being in (% predicted) respectively 5.3 (95% CI 0.6-10.0) and 4.4 (95% CI 0.1-8.7), (P < 0.05). The airway responsiveness (PD15) did not differ significantly, but the improvement from baseline to 3 years was significantly greater in the IG, being 0.40 dose steps (95% CI 0.05-0.75) (P < 0.05). HRQOL scores did not differ significantly. The risk for sickness day was less in the IG with a RR of 0.6 (95% CI 0.50-0.69) (P = 0.000) and among patients who used the PEF meter. The compliance was similar in both groups when measured by the PEF based self-management. There was no statistically significant difference in costs, although there was a consistent tendency for lower costs in the intensive programme. The intensive programme was more effective in terms of FEV1, PEF and improvement in PD15 and equally effective in terms of other lung functions and HRQOL, but there was no clear difference in the costs. PMID- 10464895 TI - Mechanisms proposed whereby beta-blockers might worsen obstructive sleep apnoea (OSA) PMID- 10464896 TI - Re: Ventilation in a Birmingham intensive care unit 1993-1995: outcome for patients with chronic obstructive pulmonary disease. PMID- 10464897 TI - Corticosteroid treatment of asthma: now at the crossroads. PMID- 10464898 TI - Chronic bronchitis and chronic obstructive pulmonary disease: Finnish National Guidelines for Prevention and Treatment 1998-2007. AB - 1. A national recommendation for the promotion of prevention, treatment and rehabilitation in relation to chronic bronchitis and COPD from 1998 to 2007 has been prepared on the basis of extensive collaboration by order of the Ministry of Social Affairs and Health. The Programme needs to be revised as necessary, because of rapid developments in medical knowledge, and in drug therapy in particular. 2. COPD is a disease characterized by slowly progressing, irreversible airways obstruction. Over 5% of the population suffer from symptomatic forms of the disease. It is estimated that a further 5% of the population may suffer from latent COPD. Most patients (75%) suffer from mild forms of the disease. The disease is often preceded by chronic bronchitis. A total of 400,000 Finns suffer from chronic bronchitis or COPD. Occurrence of these diseases in future will be particularly affected by decreased smoking by men, increased smoking by the young and by women, and aging of the population. 3. In 1997, the annual treatment costs of chronic bronchitis and COPD were estimated to be FIM 1.5 thousand million, total costs FIM 5 thousand million. Without intensification of measures to prevent and treat the diseases, costs will rise significantly. Costs arising from severe COPD (5% of patients with COPD) account for roughly 65% of costs overall and are primarily related to hospitalizations. 4. The goals of the Programme for the Prevention and Treatment of Chronic Bronchitis and COPD are as follows: (a) to decrease the incidence of chronic bronchitis; (b) to ensure that as many patients as possible with chronic bronchitis recover; (c) to maintain capacity for work and functional capacity of patients with COPD; (d) to reduce the percentage of patients with moderate to severe COPD; (e) to decrease the number of hospitalization days of COPD patients by 25% overall; and (f) to decrease annual costs per patient. 5. The following means are suggested for achieving the goals: (a) reduction in smoking; (b) reduction in work-related and outdoor air pollutants and improvement of quality of indoor air; (c) enhancement of knowledge about risk factors and treatment of the diseases is in key groups; (d) promotion of early diagnosis and active treatment, in smokers in particular; (e) improvement of guided self-care; (f) early commencement of rehabilitation, individual planning and implementation, primarily as an element in treatment; and (g) encouragement of scientific research. 6. COPD and exacerbation of its symptoms can be prevented through choices relating to life habits, such as not smoking, maintaining good general condition, and protection against exposure to dusts. The Programme gives examples of such measures and appeals to various authorities and voluntary organizations to increase their cooperation. Preventive methods should be individualized, and based on due consideration. 7. Chronic bronchitis and COPD should be diagnosed at early stages, and treated appropriately from the outset. Treatment consists of: (a) treatment according to causes, such as stopping smoking and work hygiene; (b) early rehabilitation such as patient education and guided self-care: (c) drug therapy; (d) hospital treatment; and (e) rehabilitation. 8. The hierarchy of referrals in the treatment of COPD should be revised to accord a greater role to the primary health care sector. Good exchanges of information and cooperation between the primary health care and specialized medical care sectors will all be necessary if this hierarchial model is to have the desired effect. 9. Hospital districts and health centres should ensure that different levels of the health care system are capable of fulfilling the tasks assigned to them appropriately. One specialist in each hospital district should be given charge of prevention and assembly of know-how relating to treatment, and of quality of treatment at regional level. (ABSTRACT TRUNCATED) PMID- 10464899 TI - Compliance with LTOT and consumption of mobile oxygen. AB - The aim of the study was to investigate compliance with long-term oxygen therapy (LTOT), the consumption of mobile oxygen and factors that might influence on these measures. The study included 182 patients with LTOT and 125 patients answered a questionnaire on daily activities. Information on arterial gas tension, lung function, and diagnosis was obtained from the medical file. Data on oxygen systems and the actual usage of oxygen were obtained from the oxygen suppliers. Seventy-six per cent of the patients had a concentrator and 35% had both stationary and mobile oxygen. Good compliance (use of oxygen > 15 h day-1) was seen in 65%. Of the tested factors, only mobile oxygen had an influence on the effective usage. Outdoor activity was reported in 65% of the patients. Only 48% of these patients had mobile units, while 21% of the home-bound patients had mobile units. Of the patients with mobile oxygen, only 39% used their mobile unit > 2 h week-1. Poorer usage of stationary oxygen was observed in patients with outdoor activity (2.4 hours day-1 less). Although only 16 (13%) patients reported a usage less than 15 h day-1, the actual consumption was less than 15 h day-1 in 29%. In conclusion, patients on LTOT tended to overestimate their oxygen usage. Acceptable compliance was observed in 65% of the patients. Only mobile oxygen had a significant impact on the overall compliance. As more patients had outdoor activities than expected, and as these activities had a negative impact on the usage of stationary oxygen, more attention must be paid on detecting outdoor activities. However, as fewer than half of the patients with outdoor activity and mobile oxygen used their mobile systems, there is a need for measures to improve compliance with mobile oxygen. Using lists referring to the actual usage and discussing the usage with the patients may improve the compliance and cost benefit. PMID- 10464900 TI - Lactate dehydrogenase isoenzyme analysis for the diagnosis of pleural effusion in haemato-oncological patients. AB - This study aimed to evaluate the utility of the pleural fluid lactate dehydrogenase (LDH) isoenzyme algorithm for the differential diagnosis of pleural fluid in patients with haematological malignancies. Twenty consecutive haemato oncological patients with pleural effusion, hospitalized in the Haematology Department during a 2.75-year period, were prospectively and independently evaluated for the cause of effusion by standard methods for the LDH isoenzyme algorithm. The causes of the pleural effusions established during the standard evaluations were compared to the results obtained from the LDH isoenzyme algorithm. Following the standard evaluation, the pleural effusion was attributed to congestive heart failure in one patient, to infection in six, to the underlying malignancy in 12 and to concomitant congestive heart failure and malignancy in one. LDH isoenzyme analysis correctly predicted the cause of pleural effusion in 18 patients (positive predictive value 90%). In haemato oncological patients, the pleural fluid LDH isoenzyme pattern may be helpful in the differential diagnosis of the most common causes of pleural effusion. PMID- 10464901 TI - Peak inspiratory flow through Turbuhaler in chronic obstructive airways disease. AB - Many patients with chronic obstructive airways disease (COAD) receive therapy by the inhaled route. This study was performed to assess whether patients with severe COAD could generate sufficient peak inspiratory flow (PIF) through Turbuhaler (Astra, Sodertalje) to operate it effectively. One hundred patients (45 men, 55 women, mean age 69.1 years) with COAD (mean (SD) duration 17.7 (16.3) years) and peak expiratory flow (PEF) < or = 200 l min-1 or forced expiratory volume in 1 sec (FEV1) < or = 1 litre were studied. A series of randomly assigned inspiratory and expiratory lung function tests were contiguously performed, using portable spirometers, within 48 h of a screening visit. An empty Turbuhaler was used in the study. The patients' normal medication was not restricted. Sixty-six patients were previous smokers, eight occasional smokers, 19 habitual smokers and seven had never smoked. Mean (SD) FEV1 was 0.7 (0.2) 1 and mean PEF was 182 (68) l min-1. All patients were able to generate PIF through Turbuhaler (PIF-T) of 28 l min-1 (mean 53; range 28-78 l min-1). Eighty-three patients generated PIF-T of > or = 40 l min-1. PIF-T correlated with PIF without Turbuhaler (r = 0.35), PEF (r = 0.3), FEV1 (r = 0.2) and forced vital capacity (FVC) (r = 0.23) although the relationships were too weak to be used to predict PIF-T. The results suggest that patients with severely limited lung function caused by COAD can operate Turbuhaler effectively. PMID- 10464902 TI - The safety and success rate of sputum induction using a low output ultrasonic nebuliser. AB - Induced sputum differential cell counts have been advocated as a method of non invasively assessing airway inflammation in asthma and other airway diseases. Since sputum induction usually involves delivering hypertonic saline via a high output ultrasonic nebulizer there have been concerns about its safety in asthma. There are relatively little data on the effects of sputum induction in large numbers of patients. We have examined the success rate and effect of sputum induction on forced expiratory volume in 1 sec (FEV1) in 100 inductions performed on 79 patients using a low output nebulizer. Thirty-seven patients had asthma, 29 had miscellaneous conditions (mainly chronic cough) and 13 were subjects without respiratory symptoms. Sputum was induced 10 min after 200 micrograms of inhaled salbutamol by sequential 5-min inhalations of 3, 4 and 5% saline delivered via a Fisoneb ultrasonic nebulizer and FEV1 was measured after each inhalation. Sputum induction resulted in a sample suitable for analysis in 92% of asthmatics, 90% of those with miscellaneous conditions and 100% of normal subjects. The mean (SEM) maximum per cent fall in FEV1 was 5.4% (0.1), 4.3%, (1.0) and 2.6% (1.1) in subjects with asthma, miscellaneous conditions and in asymptomatic subjects respectively. Only 13 inductions resulted in a > 10% fall in FEV1, and only three of these resulted in a > 20% fall. The maximum per cent fall in FEV1 did not correlate with baseline FEV1 % predicted (r = -0.17), the log sputum eosinophil count (r = -0.12), or the methacholine PC20 (r = -0.14). We conclude that sputum induction using a relatively low output ultrasonic nebulizer with premedication with salbutamol is successful and safe in the majority of patients with asthma and other airway conditions. PMID- 10464903 TI - Environmental asbestos exposure and malignant pleural mesothelioma. AB - Asbestos-related benign and malignant pleural diseases are endemic in some rural parts of central Turkey because of environmental exposure to asbestos fibres. We report here epidemiological data on 113 patients with diffuse malignant pleural mesothelioma (DMPM) diagnosed in our clinic in Eskisehir, located in central Turkey. Of the 113 patients, 59 were men and 54 women (male:female ratio = 1). Ninety-seven patients (86%) had non-occupational asbestos exposure; all were living in villages. Their mean age was 56 years. As the patients had been exposed to asbestos from birth, the latency period was equivalent to the age of the patients. Twenty-eight patients (29%) had lived in villages their entire lives. The other 69 (71%) had been born in a village but migrated to the city or had given up white-soil usage for various reasons. The mean exposure time was 55 years for those with a long exposure period and 25 years for those with a short exposure period, but there was no significant difference between the age of the disease appearance for both groups (55 and 56 years, respectively). Thus, the latency time of mesothelioma due to environmental exposure to asbestos was longer than that due to occupational exposure, but independent of the length of exposure. Soil samples from 67 villages were analysed, comprising a population of 10,120 villagers. Tremolite and some other types of asbestos were found. In conclusion, DMPM in our region is due to mainly to environmental exposure to asbestos. The risk is substantial as a large proportion of the villagers are exposed. After smoking, asbestos exposure is one of the most serious health hazards in our rural population. PMID- 10464904 TI - Survey of knowledge of simple pulmonary function tests (PFTs) amongst trainee doctors in England. AB - The understanding of and interpretation skills for simple pulmonary function tests (PFTs) are increasingly required in the everyday practice of medicine. This knowledge was evaluated amongst the trainee hospital doctors in the north-east of England. Three-hundred questionnaire sets were sent in the post to the relevant junior doctors working in the north-east of England and sixty-nine completed sets were returned: a raw response rate of 23% and a corrected response rate of 27%. There were deficits in knowledge and confidence in most tests except for those which were most commonly used, and this was noted across the different grades of junior doctors. The respondents were aware of the deficiency and most would like further teaching and input during their hospital career. An appropriate plan for improvement should be formulated and implemented. PMID- 10464905 TI - Midazolam sedation to produce complete amnesia for bronchoscopy: 2 years' experience at a district general hospital. AB - Patients may find bronchoscopy without sedation unpleasant. There is some evidence that patient satisfaction correlates with amnesia for the procedure. For several years we have used doses of midazolam sufficient to put patients lightly asleep hoping to produce complete amnesia. We looked at practical aspects of this technique over a 2-year period. We studied 337 consecutive patients. They were 219 men and 118 women of mean age 63 +/- 12.4 (SD). Sixty-seven patients were aged 75 years or over and the eldest was 86. Sixty-three patients were already hospital inpatients but the remainder were seen as day cases. Midazolam was given by slow i.v. injection over several minutes until the patient was judged to be lightly asleep. Patients were given supplemental oxygen (3 l min-1) and monitored by ECG and pulse oximetry. A note was made of the time at which they awakened, defined as when nursing staff felt the patients were awake enough to have a cup of tea and toast. Patients were asked if they had any memory of the procedure both on awakening and when seen a few days later to discuss the results. The procedures were carried out in a well-staffed Day Case Unit with a recovery area. The mean dose of midazolam used was 10.8 mg (mean +/- SD = 0.16 +/- 0.095 mg kg 1). The midazolam was given over a median of 4 min (range 1-15 min). Patients took 59 +/- 45 min (mean +/- SD) to wake up. Twenty-eight patients were given flumazanil to reverse the sedation (11 for concern over bleeding following biopsies, three for desaturation during and three after procedure, four as they were frail, two as they were restless, two as they were hypotensive after procedure and three for miscellaneous reasons). Only nine patients could remember any part of the procedure. Incremental doses of midazolam given slowly until patients are lightly asleep almost invariably produce complete amnesia for bronchoscopy. This is a safe technique but patients need careful monitoring and may require reversal of sedation with flumazanil. PMID- 10464906 TI - Inhaled beclomethasone dipropionate symposium. PMID- 10464907 TI - [Comparative evaluation of ACE inhibitors: which differences are relevant?]. AB - ACE inhibitors are well established in the treatment of arterial hypertension, heart failure and diabetic and/or hypertensive nephropathy with albuminuria. The important trials for the various indications are briefly discussed. In Switzerland 11 ACE inhibitors are available for clinical use, differing mainly in their pharmacokinetic and pharmacodynamic properties. The characteristics of practical relevance regarding oral bioavailability, elimination mechanisms and half-life, as well as the necessary dosage modifications in patients with renal, hepatic and cardiac failure, are presented. All ACE inhibitors except captopril and lisinopril are administered as prodrugs. The bioavailability among ACE inhibitors varies widely with a range from 11% (trandolapril) to more than 60% (captopril). The great majority of ACE inhibitors are eliminated predominantly through the kidneys. However, benazepril, fosinopril, ramipril, spirapril and trandolapril also have a hepatic (metabolic) route of elimination. Since half life varies from 1 h (captopril) to 30 h (spirapril) we drew up, for simplicity, a table of 3 groups with short, medium and long t1/2. In renal insufficiency dose adjustment is required only below a creatinine-clearance level of 30 ml/min. These dosage reductions are not required in liver diseases, but renally excreted drugs such as lisinopril should be preferred. Treatment with ACE inhibitors in severe heart failure should be initiated carefully, with low doses and concomitant diuretic treatment added or maintained. Most common adverse effects of ACE inhibitors are hypotension, cough, hyperkalaemia and renal failure. Less frequent adverse effects are angioedema, bone marrow suppression and also foetal damage. Thus, ACE inhibitors are contraindicated in pregnancy. PMID- 10464908 TI - [Prevention of post-transfusion cytomegalovirus infection: recommendations for clinical practice]. AB - Based on a review of the literature, guidelines for the clinical use of cytomegalovirus (CMV) seronegative cellular blood products are presented. The clinical fields of application include obstetrics and foetal, neonatal, and transplantation medicine. Seronegative blood products are indicated for the transfusion of pregnant women, foetuses, and neonates until the 3rd month of life. They are also indicated in patients undergoing transplantation, if the donor and the recipient are both seronegative for CMV. This indication is extended to all transplanted patients with unknown CMV serology, and for all patients after lung transplantation. Finally, deleucocyted blood products are not equivalent to CMV seronegative blood products, but their use can be an acceptable alternative in the event of shortage of CMV seronegative blood products. PMID- 10464909 TI - [Intermittent paraparesis as manifestation of a bilateral chronic subdural hematoma]. AB - Chronic subdural haematomas are mainly related to slight or moderate head trauma with consecutive lesion of bridge or cortical veins and bleeding in the subdural space. Further predisposing factors are known impairment of coagulation (coagulopathies, treatment with anticoagulants, alcohol abuse), risk factors for degenerative disease of the arteries (diabetes mellitus, arterial hypertension), and development of pressure gradients (hydrocephalus, epileptic seizures, lumbar puncture, CSF drainage and cerebral atrophy). Chronic subdural haematomas appear bilaterally in 20 to 25% of cases. We report on a 69-year-old male with a 4-day history of intermittent, proximal, painless paraparesis (BMA grade M2-5) without a trigger event. Sensibility was normal in all qualities and vigilance was not disturbed. Computed tomography of the neurocranium revealed a bitemporally located chronic subdural haematoma with extension to parietal on both sides. Trepanation was performed over the tuber parietale and temporoparietally on both sides, with release of 150 ml fluid. The neurologic deficits regressed totally within 12 hours postoperatively. To the best of our knowledge, we are the first to describe the clinical paradox of intermittent, painless paraparesis with preserved sensibility and without disturbances of vigilance, as manifestation of a chronic subdural haematoma possibly leading to impairment of cerebral blood flow in the area of the middle cerebral artery. Small changes in systemic blood pressure lead to changes in cerebral perfusion pressure due to vessel compression by the haematoma, thus explaining the intermittent character of the clinical presentation. PMID- 10464910 TI - [Present position in the prevention and therapy of NSAID-induced ulcers]. AB - The administration of nonsteroidal anti-inflammatory drugs (NSAIDs) is frequently associated with injury to the gastroduodenal mucosa and leads in approximately 1.5% of patients to severe complications such as haemorrhage or perforation. The risk of serious upper GI complications is increased in patients > 65 years with a previous history of peptic ulcer disease or gastrointestinal haemorrhage, concomitant steroid use and significant cardiovascular comorbidity. Previous studies have shown that misoprostol is effective in reducing the incidence of gastric and duodenal ulcers as well as serious gastrointestinal complications. Recently, four large clinical trials have demonstrated that omeprazole is effective in preventing and treating NSAID-induced ulcers. Omeprazole when compared to misoprostol was equally effective in preventing gastric ulcers and more effective in duodenal ulcers. For treatment of gastric and duodenal ulcers, omeprazole was more effective than misoprostol and ranitidin. Prophylaxis of NSAID-induced ulcers should be administered in all patients with several risk factors for serious gastrointestinal complications. PMID- 10464911 TI - [Refractory inguinogenital dermatosis]. PMID- 10464912 TI - [Basic components in the development of research proposals in health sciences]. AB - Any investigation in the health sciences should have a plan or proposal to answer a specific research question. This document, designed for persons preparing for or already pursuing a research project, describes the conceptual framework related with a research proposal, from the study design until its implementation. The objective of this document is to summarize different theoretical aspects that should be considered in a research project, including pertinent references that might be used to deepen the discussion. First, a series of definitions related to the scientific method are presented and then the processes to carry out a research proposal including common epidemiologic designs are illustrated. In addition, a series of research questions that can serve as a guide to perform a critical analysis of published papers are included. Finally, an interdisciplinary group integrated by health professionals with the clinical expertise and health professionals with the necessary skills in biostatistics and epidemiology to undertake a sound scientific research is recommended. PMID- 10464913 TI - Ion exchangers in radioactive waste management. Part XI. Removal of barium and strontium ions from aqueous solutions by hydrous ferric oxide. AB - The uptake of Ba(II) and Sr(II) by hydrous ferric oxide (HFO) was studied as a function of contact time, concentration, temperature and pH of the respective adsorptive solutions employing the 'radiotracer technique'. The uptake of both the ions was found to increase with the increase in concentration, temperature and pH of the adsorptive solutions. Concentration dependence data fitted well to the Freundlich adsorption isotherm over the entire range of concentration (10(-2) 10(-7) M) and the uptake process followed first-order rate kinetics. The desorption experiments demonstrate the irreversible nature of the uptake process, however, in the presence of H+ ions, i.e. on acidification, an appreciable amount of metal ions were removed in the bulk solution. The radiation stability of hydrous ferric oxide towards the uptake of Ba(II) and Sr(II) was also examined by using samples of hydrous ferric oxide irradiated by neutrons and gamma-rays prior to be employing as sorbents. The presence of some mono- and divalent co-ions along with the studied ions suppressed their removal appreciably. PMID- 10464914 TI - An automated radiosynthesis of 2-[11C]thymidine using anhydrous [11C]urea derived from [11C]phosgene. AB - 2-[11C]Thymidine has been produced from [11C]methane via [11C]phosgene and [11C]urea. Anhydrous [11C]urea was prepared from [11C]phosgene by reaction with liquid ammonia. This novel approach avoids the problems associated with the synthesis of anhydrous [11C]urea from [11C]cyanide. A fully automated system based on a modular approach and under PLC control has been developed. The system provides 2-[11C]thymidine reliably and reproducibly for clinical PET studies. The radiosynthesis takes 45-50 min from [11C]methane and the average yield was 1.5 3.3 GBq (40-90 mCi). The specific radioactivity was typically in the range 29.6 51.8 GBq mumol-1 (0.8-1.4 Ci mumol-1) at EOS corresponding to 6-12 micrograms of stable thymidine. The radiochemical yield of 2-[11C]thymidine was ca. 14% from [11C]methane. PMID- 10464915 TI - Improved radioiodination of 1,2-dipalmitoyl-3-IPPA via a tributyltin intermediate. AB - 1,2-Palmitoyl-3-[15-(4-iodophenyl)pentadecan-3-oyl]-rac-glycerol (MIPAG) is a new agent for the clinical evaluation of pancreatic lipase activity and has demonstrated promise in preliminary clinical studies with patients affected with pancreatic insufficiency. Iodine-131-MIPAG was initially prepared via thallium iodide displacement. Because of the need for a simple method which is amendable for the routine clinical use of MIPAG we have investigated the preparation and radioiodination of MIPAG utilizing the tributyltin precursor, 1,2-palmitoyl-3-[15 (4-tributylstannylphenyl)pentadecan-3-oy l]-rac-glycerol (TBT-MIPAG, 2). Compound 2 was prepared via the condensation of 1,2-palmitoyl-rac-glycerol with 15-(4 tributylstannylphenyl)pentadecanoic acid (TBT-PPA) prepared from 4 bromophenylacetylene. Electrophilic radioiodination using peracetic acid with sodium iodide-125 in ethanol at 80 degrees C for 60 min afforded I-125-MIPAG in 65.9% (+/- 11.5%) yield and radiochemical purity of 94% (+/- 3.0%) after C-18 Sep Pak purification (n = 6). This improved method for radioiodination utilizing TBT MIPAG now provides radioiodinated MIPAG for routine clinical evaluation. PMID- 10464916 TI - Pu-241 in samples of forest soil from Poland. AB - 241Pu activity measurements in coniferous forest soil samples from Poland are presented. The results were obtained in two ways: by alpha spectrometric remeasurements of the plutonium sources 3-4 years after their preparation (i.e. by the 241Am ingrowth) and by direct measurements of 241Pu using liquid scintillation (LS) spectrometry. Both methods gave consistent results. The maximum observed activity concentration obtained by the 241Am ingrowth was (254 +/- 43) Bq/kg, and by direct measurements it was (284 +/- 31) Bq/kg (the same sample, activities calculated for May 1, 1986). Enhanced levels of 241Pu were observed in all samples from the farthest north-eastern Poland. The estimated 241Pu maximum deposition from Chernobyl fallout in this area (sum of deposition for two examined layers of one site) was (1.025 +/- 0.110) kBq/m2. This will result after 70 years in an additional 241Am activity of about (30.1 +/- 3.2) Bq/m2. The average ratio for 241Pu to total 238,239,240Pu was of the order of 25. The obtained average Chernobyl ratios for 241Pu to 239,240Pu were about 86, and those for 241Pu to 238,239,240Pu were 56. PMID- 10464917 TI - Application of a new method of searching for geochemical changes related to seismic activity. AB - 222Rn in groundwater at three out of four New Zealand sites changed in a statistically significant way in response to increased monthly coseismic events M2.0-M5, 14-100 km away, particularly events normal to a nearby large faultline. delta 18O(H2O) at one site showed a coseismic negative relationship to seismicity the same day. Of other parameters at the four sites (CO2, CH4/N2, H2/N2, delta 18O(H2O), delta 13C(CO2), delta 13C(CH4), D/H, Ar/N2), only the latter showed some consistent sensitivity to seismicity, mainly as after-effects. There was no observable threshold strain value. The 9000 events studied gave considerable statistical power, but correlations useful for seismicity prediction were few and doubtful. The frequent negative relationships may indicate the closing of rock cracks by strain, hence reduced gas flow. PMID- 10464918 TI - Determination of 222Rn in natural water samples from health resorts in the Sudety mountains by the liquid scintillation technique. AB - A method involving a Wallac 1414 WinSpectral alpha/beta liquid scintillation counter for determining 222Rn in aqueous samples is described. Samples were collected from 23 springs and 4 taps in health resorts of the Klodzka valley in the Sudety mountains in Poland. Half of the investigated spring water samples were radon enriched with an activity concentration higher than 74 Bq/l. In the tap waters the radon concentration level is very low or below the lower limit of detection. Owing to the statistical nature of radioactive decay the uncertainty of the measurement was determined as the standard deviation of 222Rn activity. The method introduced is fast and simple and does not require a chemical sample preparation procedure. PMID- 10464919 TI - The measurement of U, Th and K concentrations in building materials. AB - A shielded high purity germanium detector has been used to measure the abundance of U, Th and K in aggregate, sand and cement. The measurements were performed in order to demonstrate that the materials contained less than 10 ppm U and less than 20 ppm Th and were therefore suitable for the construction of a low background shielding enclosure. A particular feature of this work is the simplicity of the absolute efficiency calibration. In brief the relative U and Th contents of each sample were derived by first determining the relative detection efficiency curve for each sample using the known relative gamma-ray emission probabilities of the U- and Th-decay series lines from the radionuclides naturally present. To obtain the true activities, the relative efficiency curves for each sample were scaled so that they passed through the absolute efficiency value measurement at 1461 keV using a sample containing KCl bought as an analytical reagent and requiring no special radiological handling precautions. A small experimentally estimated correction was applied for the difference in self absorption between the KCl and building materials due to the differences in material density. The 40K content of the unknown samples was obtained directly from the density adjusted KCl calibration. The measurements are believed to be accurate to approximately +/- 5% absolute. PMID- 10464920 TI - Urinary excretion measurements for the assessment of body burden of radiocaesium in man: differences between potassium and creatinine normalisation. AB - Relationships between 24-h urinary excretion and body burden of 137Cs obtained in a south-Swedish population, together with data from the literature, were applied to urine samples collected in 1994-95 from adult subjects living in Russia, in order to estimate their average body burden of 137Cs. Estimates obtained through creatinine normalisation of the 137Cs content in the Russian urine were a factor of 2 higher than estimates obtained through potassium normalisation. The accuracy of both these normalisation methods is discussed, as well as the influence of the difference in dietary habits between the two populations. PMID- 10464921 TI - The standardization of 62Cu and experimental determinations of dose calibrator settings for generator-produced 62CuPTSM. AB - The Positron Emission Tomography agent 62Cu Pyruvaldehyde Bis(N4 methyl)thiosemicarbazone (PTSM) has been standardized by the National Institute of Standards and Technology (NIST) using 4 pi beta liquid scintillation counting with 3H-standard efficiency tracing. Using a measurement model developed at NIST for the determination of experimental dose calibrator dial settings for short lived radionuclides, the correct dial settings for 62CuPTSM in a clinically useful geometry (a 35 ml plastic syringe containing 33 ml of solution) as well as the standard NIST 5 ml glass ampoule have been elucidated. This measurement model is fully described, as is the treatment of associated uncertainties. The correct instrument settings for the NIST Capintec CRC-12 re-entrant ionization chamber ('dose calibrator') were found to be '499 +/- 6' and '489 +/- 8', respectively, for the syringe and ampoule geometries. The stated uncertainties are expanded (k = 2) uncertainties. The expanded uncertainties in the measured activity arising from these new dial settings are +/- 0.8% and +/- 1.4%, respectively, for the syringe and ampoule. The measured activities using these settings are lower than those obtained from the manufacture's recommended setting of '448' by 9.7% for the syringe and 7.7% for the ampoule, and underscore the need for experimental verification of the dose calibrator settings for each radionuclide and each geometry used in the clinic. PMID- 10464922 TI - Determination of natural radioactivity in building materials used in the Rawalpindi/Islamabad area by gamma-ray spectrometry and instrumental neutron activation analysis. AB - Samples of sand and manufactured building materials collected from the Rawalpindi/Islamabad area have been analyzed for the primordial natural radionuclides 238U, 232Th and 40K using gamma-ray spectrometry. The uranium and thorium contents have also been determined by instrumental neutron activation analysis (INAA). The specific activities of these radionuclides in the samples are compared with those of the world averages for soil. The comparison shows that, of the nine samples analyzed, only the three brick samples have higher activities than the world averages for soil. The specific activities in these materials, having a radium equivalent activity of less than 370 Bq kg-1, when evaluated for radiological effects show that all materials meet the external gamma-ray dose limitation of 1.5 mSv y-1. The gamma-spectrometric and INAA techniques complemented each other well in this study. PMID- 10464923 TI - Absorbed doses to patients from 191Pt-, 193mPt- and 195mPt-cisplatin. AB - Cisplatin, a chemotherapeutic drug, can be synthesized using radioactive platinum and then used for pharmacokinetic studies and tumor imaging. We have calculated the absorbed doses to various organs and tissues as well as the effective doses from 191Pt-, 193mPt- and 195mPt-cisplatin after administration to humans for diagnostic purposes. Liver was the organ that received the highest absorbed dose. The effective dose from 191Pt-, 193mPt- and 195mPt-cisplatin was 0.10 +/- 0.02, 0.17 +/- 0.04 and 0.23 +/- 0.05 mSv/MBq respectively. PMID- 10464924 TI - Uranium in mining water of kaolin open pit in Zarow (Lower Silesia); methodology of determination and genetic remarks. AB - In this paper, a method of determination of uranium 238 and 234 in mining waters of Andrzej kaolin open pit in Zarow (Lower Silesia) is presented. The method is based on independent measurements of alpha and beta radiation intensities by means of a liquid scintillation spectrometer alpha/beta. The initial volume of water sample was 3 dm3, then it was diminished by chemical preparation to 6 cm3, and then 12 cm3 of scintillator was added. The lower limit of detection (for the measurement time of 8 h) for both 234U and 238U amounted to 0.02 Bq/dm3. For determination of the uranium content in ferruginous sediments precipitating from mining waters of the above-mentioned open pit, gamma ray spectrometry was used. The obtained results may be viewed as a contribution to studies on anomalous uranium concentration within this kaolin deposit. The elevated uranium content, in comparison with its average concentration in the Earth crust, is characteristic for parent rocks of Andrzej kaolin deposit, which are granitoids of Strzegom-Sobotka massif. In connection with it, the high uranium content can be observed not only in kaolin and weakly kaolinised granitoids from the deposit in question, but also in mining waters genetically related with them. PMID- 10464925 TI - Fallout beryllium-7 as a tracer in soil erosion investigations. AB - Recent developments in the use of the environmental radionuclides 137Cs and 210Pb for documenting medium-term soil erosion rates must be seen as an important advance. However, measurements of these radionuclides provide estimates of medium term (i.e. ca. 45 or 100 years) soil redistribution rates and there is a need to explore the potential for using complementary radionuclides, to estimate erosion rates associated with individual events or short periods. Beryllium-7 (7Be, t0.5 = 53 days) has the potential to fulfil these requirements. This contribution presents some preliminary results from an investigation undertaken in a field near Crediton, UK, which successfully demonstrates the potential for using 7Be as a tracer in soil erosion investigations. PMID- 10464926 TI - In vitro studies on the effects of some anthelmintics on Cotylophoron cotylophorum (Digenea, Paramphistomidae): a structural analysis. AB - The effects of the anthelmintics praziquantel (PZQ), levamisole (LEV), mebendazole (MBZ), fenbendazole (FBZ) and albendazole (ABZ), on the morphology and the histology of a digenetic trematode, Cotylophoron cotylophorum, were studied. Scanning electron micrographs of the drug-treated worms revealed that PZQ was the most effective drug inducing surface damages to a great extent. The parasite exposed to PZQ for 6 h, showed smaller blebs on the oral sucker region as well as on the sensory papillae. These blebs enlarged in size after 24 h and ruptured after 30 h of exposure. The worms treated with LEV showed a few smaller blebs on the ventrolateral margin. In MBZ- and FBZ-treated worms the blebs appeared between the oral and genital sucker after 6 h of incubation. The changes were not apparent in the ABZ-treated worms. PMID- 10464927 TI - An assessment of human performance in stabbing. AB - Stab-resistant body armour is now becoming a standard item of equipment for police officers in the United Kingdom. In the UK these are usually required to have a stab resistance as specified by the Police Scientific Development Branch KR42 standard [G. Parker, PSDB Stab Resistant Body Armour Test Procedure, Police Scientific Development Branch, Publication No 10/93, 1993]. There are several other test standards, all of which specify that body armour must resist penetration by a specific blade type delivered at a specific energy level or range of levels. However, the actual range of energy levels specified varies over almost an order of magnitude and the basis for these levels is not clearly defined. This paper describes tests to determine the energy range and characteristics of stabbing actions that might be directed against stab resistant body armour by an assailant. The energy and velocity that can be achieved in stabbing actions has been determined for a number of sample populations. Volunteers were asked to stab a target using an instrumented knife that measured the axial force and acceleration during the stabbing. The maximum energy obtained in underarm stabbing actions was 64 J whilst overarm stabbing actions could produce 115 J. The loads produced on contact with the target often approached 1000 N. PMID- 10464928 TI - The use of alcohol and other psychoactive substances by victims of traffic accidents in Greece. AB - Alcohol is one of the main causes of traffic accidents worldwide. Its use decreases significantly the driving ability of an individual as well as the reflexes of a pedestrian increasing in this way the possibilities of their involvement in motor-vehicle accidents. The above possibilities are increased when a psychoactive substance has been taken in combination with alcohol due to their synergistic effect. The Laboratory of Forensic Medicine and Toxicology of the University of Athens is authorized to perform the toxicological investigation of traffic accidents that happen in the major part of Greece. The findings of the toxicological analyses that were performed in our laboratory during the years 1995 to 1997 are classified according to the alcohol concentration and the kind of psychoactive substance detected as well as to the gender and the age of the victims, drivers or pedestrians. The results of the above classification are evaluated and the conclusions regarding alcohol, drugs and traffic accidents in Greece are discussed thoroughly. PMID- 10464929 TI - DNA typing for identification of some species of Calliphoridae. An interest in forensic entomology. AB - To determine precisely post mortem interval, larvae and puparium species found on a corpse have to be identified. Among more than 200 cases examined at the entomology department of the Institut de Recherche Criminelle de la Gendarmerie Nationale, two-thirds concerned corpses less than one month old. Therefore, insects from first and second screwworms are the most frequently found [1]. Some species commonly found in France, such as different Lucilia and Calliphora vicina Robineau-Desvoidy, are easily identifiable at an adult stage, but are almost impossible to differentiate at immature stages when only fragments of puparium or necrosed first instar larvae are available. For this reason, an easy and objective method of identification was thus searched by genetic analysis of these insects. Sequencing of partial gene of sub unit I of cytochrome oxydase has been used to predict restriction sites. Restriction enzyme cleavage of PCR products with Dde I allowed us to differentiate these species. PMID- 10464930 TI - What happens in freezing bodies? Experimental study of histological tissue change caused by freezing injuries. AB - In order to evaluate histological features of freezing damages to human tissue after death, we froze samples of liver and heart tissue to temperatures of -12 degrees C, -28 degrees C and -80 degrees C, and stored them for 24 and 72 h, respectively, at those temperatures. After thawing and routine preparation for histology, the samples were evaluated both by microscope and with an electronic image analyzer. In all cases, we found extended extracellular spaces and shrunken cells resulting from the freeze-thaw cycle. These features were more pronounced in tissues stored for longer durations. Such findings seem to be typical of tissue that has been frozen prior to examination. Two cases of dead bodies found outdoors at subzero temperatures demonstrate that formerly frozen and unfrozen tissues can be distinguished histologically. The findings are examined in relation to the fundamental laws of cryobiology. PMID- 10464931 TI - Results of collaborative study regarding the standardization of the Y-linked STR system DYS385 by the European DNA Profiling (EDNAP) group. AB - Y-chromosome linked short tandem repeat (STR) loci are inherited as a closely linked haplotype, which appears to remain stable in a given paternal lineage over many generations. In forensic cases, Y-linked STRs are particularly useful for the identification of human remains as well as in rape cases with mixed male/female stain samples. DYS385 is derived from tandemly duplicated segments of the Y chromosome thus giving rise to two fragments of variable length which do not behave like alleles but genotypes. The European DNA Profiling (EDNAP) group has carried out a collaborative exercise among 14 participating laboratories using DYS385 for typing of five unknown bloodstains and a control sample. Furthermore, population data from eight different European countries with samples sizes between 91 and 150 male individuals were collected. The results confirm previous observations that DYS385 is one of the most informative Y-linked STR loci. It could also be demonstrated that reproducible results can be obtained independently from the electrophoretic separation and detection methods used. Thus DYS385 may serve as a useful complementation to the routinely used autosomal STR systems in special cases. PMID- 10464932 TI - The detection of cotinine in hydrolyzed meconium samples. AB - To date, the screening of meconium for the determination of tobacco exposure in newborns has proven difficult. It was hypothesized that cotinine forms reversible Schiff base bonds with free amino functions on proteins, therefore, hydrolysis of meconium would be necessary for the detection of 'free' cotinine. One-hundred-and two (102) meconium samples received into our laboratory were extracted using a routine non-hydrolysis screening procedure for drugs of abuse. Separate aliquots of the specimens were hydrolyzed and re-extracted according to the same procedure. The results of the two methods were compared using a highly specific cotinine micro-plate enzyme immunoassay procedure (EIA). Of the non-hydrolyzed samples, 33% were positive for cotinine, while 79% of the hydrolyzed samples were cotinine-positive. Common drugs of abuse did not interfere with the analysis. Micro-plate EIA provides a rapid, simple and reliable screening method for the determination of cotinine in meconium following hydrolysis and extraction. In general, the meconium specimens received into our laboratory are from newborns considered to be at risk for post-natal problems due to suspected drug and/or alcohol abuse during pregnancy. PMID- 10464933 TI - Percentile charts to determine the duration of child abuse by chronic malnutrition. AB - Longstanding quantitative or qualitative under-supply of nutrition leads to weight loss and, in children, to stagnation of growth and thus to stunted growth. A comparison of the expected growth, according to percentile growth curves, with the actual body size, gives an indication as to the period of time in which malnutrition took place. The moment in which the growth curve bends off and leaves the norm is to be interpreted as the earliest begin, the moment in which the attained growth would have been achieved as the latest begin of the nutritional impairment. PMID- 10464934 TI - Determining the human origin of fragments of burnt bone: a comparative study of histological, immunological and DNA techniques. AB - In situations where badly burnt fragments of bone are found, identification of their human or non-human origin may be impossible by gross morphology alone and other techniques have to be employed. In order to determine whether histological methods were redundant and should be superseded by biomolecular analyses, small fragments of artificially burnt bone (human and non-human) were examined by quantitative and standard light microscopy, and the findings compared with newer biomolecular analyses based on identifying specific human albumin by ELISA and amplifying human mitochondrial DNA by PCR. For quantitative microscopy, reference data were first created using burnt bones from 15 human and 20 common domestic and farm animals. Measured osteon and Haversian canal parameters were analysed using multivariate statistical methods. Highly significant differences were found between values for human and non-human bone, and a canonical discriminant function equation was derived, giving a predicted correct classification of 79%. For the main study, samples of cortical bone were taken from three fresh cadavers, six human skeletons and ten freshly slaughtered animals and burnt by exposure to temperatures ranging from 800 to 1200 degrees C; charred fragments of human cortical bone from two forensic cases were also tested. Quantitative microscopy and canonical discriminant function gave the correct origin of every sample. Standard microscopy falsely assigned burnt bone from one human skeleton and one forensic case to a non-human source, but otherwise gave correct results. Human albumin was identified in five individuals, including one of the forensic cases, but mitochondrial DNA could not be amplified from any of the human bone. No false positive test results were seen with either biomolecular method; and human albumin and mitochondrial DNA were correctly identified in all unburnt control specimens. It was concluded that histological methods were not redundant and that quantitative microscopy provided an accurate and consistent means of determining the human or non-human origin of burnt bone and was more reliable than standard microscopy or the newer immunological and DNA techniques tested here. PMID- 10464935 TI - Tentative injuries to exposed skin in a homicide case. AB - A 28-year-old man stabbed both his wife and his 3-year-old son to death, before unsuccessfully attempting to commit suicide. The incident occurred against a background of marital conflict. The child's body exhibited six tentative wounds to the skin in the area of the heart, with no corresponding defects in the overlying clothing, a pattern normally seen only in suicide. Their presence can be explained by the fact that this can be considered an extended suicide, the father's motivation for the killing being comparable to that for true suicide. However, wounds of this nature can be produced in such cases only if the victim is severely limited in his ability to defend himself, here due to the superior physical strength of the father. PMID- 10464936 TI - Mitochondrial DNA regions HVI and HVII population data. AB - Data from 1393 unrelated individuals have been compiled from eight population groups: African Americans, Africans (Sierra Leone), U.S. Caucasians, Austrians, French, Hispanics, Japanese, and Asian Americans. The majority of the mtDNA sequences were observed only once within each population group (i.e., ranging from a low of 60.3% (35/58) of the Asian American sequences to a high of 85.3% (93/109) of the French sequences). Genetic diversity ranged from 0.990 in the African sample to 0.998 in African Americans. Random match probability ranged from 2.50% in the Asian American sample to 0.52% in U.S. Caucasians. The average number of nucleotide differences between individuals in a database is greatest for the African American and African samples (14.1 and 13.1, respectively), and the least variable are the Caucasians (ranging from 7.2 to 8.4). Substitutions are the predominate polymorphism, and at least 92% of the substitutions are transitions. The most prevalent transversions are As substituted for Cs and Cs substituted for As. For most population groups these transversions occurred predominately in the HVI region; however, the African, African American, and Hispanic samples also demonstrated a large portion of their C to A and A to C transversions in the HVII region (at sites 186 and/or 189). Most insertions occur in the HVII region at sites 309.1 and 315.1, within a stretch of C's. Insertions of an additional C are common in all population groups. The sequence data were converted to SSO mtDNA types and compared with population data on Caucasians, Africans, Asians, Japanese, and Mexicans described by Stoneking et al. [M. Stoneking, D. Hedgecock, R.G. Higuchi, L. Vigilant, H.A. Erlich, Population variation of human mtDNA control region sequences detected by enzymatic amplification and sequence-specific oligonucleotide probes, Am. J. Hum. Genet. 48 (1991) 370-382] using an R x C contingency table test. Differences between major population groups (i.e., between African, Caucasian, and Asian) are quite evident, and similar ethnic population groups carried similar SSO polymorphism frequencies. There were only a few SSO types that showed significant differences between subpopulation groups. The SSO data alone can not be used to describe the population genetics with complete sequence data. However, the results of the SSO comparisons are similar to other analyses, and differences in sequence data in regions HVI and HVII are greater between major population groups than between subgroups. PMID- 10464937 TI - Diatom test with Soluene-350 to diagnose drowning in sea water. AB - The authors tested the Soluene-350 method to detect diatoms in three cases of sea water drowning. The negative results obtained in all three cases prompted us to re-test the method on samples of fresh water and sea water microalgae. The experiment showed that the siliceous frustule of sea water diatoms is solubilized by Soluene-350 while that of fresh water diatoms is resistant to the treatment. The method, which is effective in cases of drowning in fresh water, should therefore be used with caution in sea water drowning. PMID- 10464938 TI - Catecholamines in urine after death. AB - Noradrenaline, adrenaline and dopamine levels were measured using high performance liquid chromatography (HPLC) and electrochemical detection and related to urine creatinine in 30 specimens from unselected autopsies. Values calculated from patients with essential hypertension were used as a reference and raised values of noradrenaline, adrenaline and dopamine were found in 76.6, 80 and 23.3%, respectively. Using the Mann-Whitney U test, there was no difference between the 11 patients with myocardial infarction and the seven who died from severe head injury or multiple trauma. The median values in mmol/mol creatinine for patients with myocardial infarction and trauma were 0.07 and 0.12 for noradrenaline and 0.012 and 0.024 for adrenaline, respectively. Physiological stress is common before death and urine noradrenaline values at autopsy overlap those found in some normal subjects and at autopsy in patients with phaeochromocytoma. PMID- 10464939 TI - The scaling of human grip configurations. AB - The many degrees of freedom of the hand and arm afford the wide range and rich adaptability of human grip configurations in action. Several classification schemes of human grip configurations have been proposed, but none is based on scaling laws of physical biology, which are well established for other categorizations of fundamental physical activities such as locomotion. This study examined the preferred human grip configurations used to displace to a new location cubes that varied systematically in length (L), mass (M), and density (ML-3). The body-scaled equation K = log L + (log M)/h (where h refers to anthropometric measures of the hand) predicted the grip configurations used to displace objects. The findings suggest that information about the dynamic scaling relation is picked up visually and organizes the many degrees of freedom of the hand-arm complex in the coordination of prehensile grip configurations. PMID- 10464940 TI - Activation of phonological codes during eye fixations in reading. AB - Two experiments addressed the issue of whether phonological codes are activated early in a fixation during reading using the fast-priming technique (S. C. Sereno & K. Rayner, 1992). Participants read sentences and, at the beginning of the initial fixation in a target location, a priming letter string was displayed, followed by the target word. Phonological priming was assessed by the difference in the gaze duration on the target word between when the prime was a homophone and when it was a control word equated with the homophone on orthographic similarity to the target. Both experiments demonstrated homophonic priming with prime durations of about 35 ms, but only for high-frequency word primes, indicating that lexicality was guiding the speed of the extraction of phonological codes early in a fixation. Evidence was also obtained for orthographic priming, and the data suggest that orthographic and phonological priming effects interact in a mutually facilitating manner. PMID- 10464941 TI - Infants' and adults' perception of scale structure. AB - Adults and 9-month-old infants were required to detect mistuned tones in multitone sequences. When 7-tone versions of a common nursery tune were generated from the Western major scale (unequal scale steps) or from an alternative scale (equal steps), infants detected the mistuned tones more accurately in the unequal step context than in the equal-step context (Experiment 1). Infants and adults were subsequently tested with 1 of 3 ascending-descending scales (15 tones): (a) a potentially familiar scale (major) with unequal steps, (b) an unfamiliar scale with unequal steps, and (c) an unfamiliar scale with equal steps. Infants detected mistuned tones only in the scales with unequal steps (Experiment 2). Adults performed better on the familiar (major) unequal-step scale and equally poorly on both unfamiliar scales (Experiments 3 and 4). These findings are indicative of an inherent processing bias favoring unequal-step scales. PMID- 10464942 TI - Change of intensity fails to produce an irrelevant sound effect: implications for the representation of unattended sound. AB - Sequences of changing sounds that are irrelevant to the task at hand disrupt serial recall appreciably even though participants are instructed to ignore the sounds. Three experiments, which compared the effect of changing token identity with that of changing intensity in the 55- to 85-dB (A) range, were conducted. Although serial recall was impaired by changes in token identity, no disruptive effects of a change in intensity were found. This was replicated using speech and nonspeech. Overall, the absence of a changing intensity effect was based on an analysis of the performance of 115 participants in a design whose power was .98. This outcome suggests that the representation of intensity in preattentive processing of auditory stimuli is somewhat different from that of other acoustic features. PMID- 10464943 TI - Robust representations for faces: evidence from visual search. AB - We report evidence from visual search that people can develop robust representations for highly overlearned faces. When observers searched for their own face versus the face of an unfamiliar observer, search slopes and intercepts revealed consistently faster processing of self than stranger. These processing advantages persisted even after hundreds of presentations of the unfamiliar face and even for atypical profile and upside-down views. Observers not only showed rapid asymptotic recognition of their own face as the target, but could reject their own face more quickly as the distractor. These findings suggest that robust representations for a highly overlearned face may (a) mediate rapid asymptotic visual processing, (b) require extensive experience to develop, (c) contain abstract or view-invariant information, (d) facilitate a variety of processes such as target recognition and distractor rejection, and (e) demand less attentional resources. PMID- 10464944 TI - Dynamic frequency change influences loudness perception: a central, analytic process. AB - Three experiments showed that dynamic frequency change influenced loudness. Listeners heard tones that had concurrent frequency and intensity change and tracked loudness while ignoring pitch. Dynamic frequency change significantly influenced loudness. A control experiment showed that the effect depended on dynamic change and was opposite that predicted by static equal loudness contours. In a 3rd experiment, listeners heard white noise intensity change in one ear and harmonic frequency change in the other and tracked the loudness of the noise while ignoring the harmonic tone. Findings suggest that the dynamic interaction of pitch and loudness occurs centrally in the auditory system; is an analytic process; has evolved to take advantage of naturally occurring covariation of frequency and intensity; and reflects a shortcoming of traditional static models of loudness perception in a dynamic natural setting. PMID- 10464945 TI - A sublexical locus for repetition blindness: evidence from illusory words. AB - Six experiments used an illusory words paradigm to demonstrate that repetition blindness (RB) in orthographically similar words affects only the words' shared letters. Rapid serial visual presentation streams of words and word fragments allowed the unique letters of the 2nd critical word to combine with a subsequent fragment to create a word, as in rock shock ell. The illusory word shell was reported 2-3 times as frequently in RB conditions as in control conditions. Further experiments ruled out letter migration, contour summation, and differences in processing load as explanations for the results. These findings are inconsistent with current proposals that orthographic RB represents similarity inhibition or lexical competition or that it reflects problems with word-level token individuation. PMID- 10464946 TI - Visual-motor recalibration in geographical slant perception. AB - In 4 experiments, it was shown that hills appear steeper to people who are encumbered by wearing a heavy backpack (Experiment 1), are fatigued (Experiment 2), are of low physical fitness (Experiment 3), or are elderly and/or in declining health (Experiment 4). Visually guided actions are unaffected by these manipulations of physiological potential. Although dissociable, the awareness and action systems were also shown to be interconnected. Recalibration of the transformation relating awareness and actions was found to occur over long-term changes in physiological potential (fitness level, age, and health) but not with transitory changes (fatigue and load). Findings are discussed in terms of a time dependent coordination between the separate systems that control explicit visual awareness and visually guided action. PMID- 10464947 TI - The effects of "neighborhood size" in reading and lexical decision. AB - The effects of neighborhood size ("N")--the number of words differing from a target word by exactly 1 letter (i.e., "neighbors")--on word identification was assessed in 3 experiments. In Experiments 1 and 2, the frequency of the highest frequency neighbor was equated, and N had opposite effects in lexical decision and reading. In Experiment 1, a larger N facilitated lexical decision judgments, whereas in Experiment 2, a larger N had an inhibitory effect on reading sentences that contained the words of Experiment 1. Moreover, a significant inhibitory effect in Experiment 2 that was due to a larger N appeared on gaze duration on the target word, and there was no hint of facilitation on the measures of reading that tap the earliest processing of a word. In Experiment 3, the number of higher frequency neighbors was equated for the high-N and low-N words, and a larger N caused target words to be skipped significantly more and produced inhibitory effects later in reading, some of which were plausibly due to misidentification of the target word when skipped. Regression analyses indicated that, in reading, increasing the number of higher frequency neighbors had a clear inhibitory effect on word identification and that increasing the number of lower frequency neighbors may have a weak facilitative effect on word identification. PMID- 10464948 TI - Extraction of information to the left of the fixated word in reading. AB - The present experiment used 2 different eye-contingent display change techniques to determine whether information is extracted from English text even when it is to the left of the currently fixated word. Preview display changes were during the 1st saccade entering the target word region, whereas postview display changes were during the 1st saccade leaving that region. Previews and postviews were either identical, related, or unrelated to the target word. "Wrong" information in the target-word region affected reading even when that information was seen only after readers were fixating to the right of that region: When readers skipped the target word, such information caused readers to regress to the target word more; when readers initially fixated the target word, such information increased "2nd-pass" processing time on the target region. The data suggest that readers often still attend to a word after it is skipped and that when readers fixate a word, they occasionally attend to the word after they have begun to fixate the next word. PMID- 10464949 TI - Phylogenetic characterization of Centipeda periodontii, Selenomonas sputigena and Selenomonas species by 16S rRNA gene sequence analysis. AB - The nearly complete 16S rRNA gene sequences for oral Gram-negative anaerobic motile bacteria, Centipeda periodontii, Selenomonas sputigena and Selenomonas species (formerly S. sputigena type strain), were determined in order to unveil their relationship to other oral motile bacteria. To determine the phylogenetic characterization of these bacteria, their 16S rRNA gene sequences were obtained and compared with those from the ribosomal sequence databases previously reported. The 16S rRNA gene sequences of these bacteria were similar to those of Selenomonas ruminantium and Schwartzia succinivorans isolated from rumens, and to Pectinatus cerevisiiphilus isolated from spoiled beer. Among oral bacteria, the nucleotide sequence analysis of these bacteria revealed high nucleotide similarity to Veillonella species, whereas low similarity to oral motile bacteria such as Campylobacter species. Phylogenetic analysis clearly confirmed that C. periodontii and two Selenomonas species were classified as relatives of a group besides Selenomonas, Schwartzia, and Pectinatus species, and not as close relatives to oral motile bacteria, such as Campylobacter species. These results suggest that such oral Gram-negative anaerobic motile bacteria are close relatives of oral bacteria. PMID- 10464950 TI - Effect of Halococcus morrhuae and Halobacterium saccharovorum on the activation of human peripheral blood lymphocytes. AB - The immunomodulator properties of two species of halophilic Archaebacteria, Halobacterium saccharovorum and Halococcus rnorrhuae, were analysed by the study of lymphocyte activation. Two methods were used to detect activation in lymphocytes, namely incorporation of the radioactive nucleotide [3H]-thymidine, and CD25 expression. H. morrhuae had a stimulatory effect on human lymphocytes, but this action was observed only with the [3H]-thymidine uptake method, whereas H. saccharovorum produced no immunomodulator effect. PMID- 10464951 TI - Catabolism of the methyl derivatives of adenosine and cytidine in staphylococci. AB - Products of 1-methyladenosine, 2'-O-methyladenosine, 2'-O-methylcytidine, and 5 methylcytidine catabolism by resting cells of Staphylococcus aureus and Staphylococcus intermedius were chromatographically separated. The methyl group in 1-methyladenosine protected the adenosine derivative from deamination by S. intermedius but it did not protect N-glycosidic bond from cleavage by S. intermedius and S. aureus. The methyl group in 2'-O-methyladenosine and 2'-O methylcytidine protected the N-glycosidic bond from cleavage by S. aureus and S. intermedius but it did not protect the adenosine and cytidine derivatives from deamination by S. intermedius. 5-Methylcytidine was converted by the common route in which 5-methylcytidine was first deaminated to ribothymidine which was cleaved to yield thymine. S. intermedius deaminated the purine and pyrimidine ribonucleosides adenosine, 2'-O-methyladenosine, cytidine, and 5-methylcytidine. Pyrimidine ribonucleosides (cytidine, 5-methyl-cytidine) were deaminated only slowly and purine ribonucleosides (adenosine, 2'-O-methyladenosine) not at all by S. aureus. PMID- 10464952 TI - Purification and characterization of an arylamine N-acetyltransferase from Lactobacillus acidophilus. AB - N-acetyltransferase from Lactobacillus acidophilus was purified by ultrafiltration, DEAE-Sephacel, gel filtration chromatography on Sephadex G-100, and DEAE-5pw on high performance liquid chromatography, as judged by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) on a 12% (w/v) slab gel. The purified enzyme was thermostable at 37 degrees C for 1 h with a half-life of 32 min at 37 degrees C, and displayed optimum activity at 37 degrees C and pH 7.0. The K(m) and Vmax values for 2-aminofluorene were 0.842 mM and 2.406 nmol/min/mg protein, respectively. Among a series of divalent cations and salts, Zn2+, Ca2+, Fe2+, Mg2+, and Cu2+ were demonstrated to be the most potent inhibitors. The enzyme had a molecular mass of 44.9 kD. The three chemical modification agents, iodoacetamide, phenylglyoxal, and diethylpyrocarbonate, all exhibited dose-, time-, and temperature-dependent inhibition effects. Preincubation of purified N-acetyltransferase with acetyl coenzyme A (AcCoA) provided significant protection against the inhibition of iodoacetamide and diethylpyrocarbonate, but only partial protection against the inhibition of phenylglyoxal. These results indicate that cysteine, histidine, and arginine residues are essential for this bacterial activity, and the first two are likely to reside on the AcCoA binding site, but the arginine residue may be located close to the AcCoA binding site. This report is the first demonstration of acetyl CoA:arylamine N-acetyltransferase in L. acidophilus. PMID- 10464953 TI - Effect of ultraviolet light irradiation and nitrosoguanidine on viability of 46 strains of Arthrinium and their antibiotic production. AB - The ability of strains of the Arthrinium genus to inhibit microbial development has been previously described. In the present work different periods of mutagenic treatment using ultraviolet light, and of nitrosoguanidine treatment, on strains of Arthrinium were investigated. With nitrosoguanidine treatment the survival rate ranged from 2.17 to 8.78%. Mutant strains were only obtained with a higher antibiotic production in comparison with the wild-strain, when the mutagenic agent was UV light. PMID- 10464954 TI - [Evolution of prostate cancer diagnostic profile over the 1994-1998 period. Efficacy of bone scanning and computerized tomography as staging methods]. PMID- 10464955 TI - [Reliability of routine cytological diagnosis in bladder cancer]. AB - OBJECTIVES: To establish the reliability of three cystopathologists for cytological diagnosis of primary bladder tumors. METHODS: Pre-operative voided urine specimens of 71 patients with bladder cancer and 55 healthy controls were retrospectively and blindly reviewed by 3 independent cytologists, and their results--positive for bladder cancer, negative or inconclusive--were compared with each other. The estimation of the interobserver agreement was calculated using the weighted kappa-coefficient (k). A multivariative analysis was carried out to identify the factors associated with the disagreement among the three observers. The sensitivity and specificity for each of the participants were calculated in order to clearly identify the origin of the disagreement, in terms of the performance of the diagnostic test in the hands of each observer. A comparison of the overall diagnostic performance was made by plotting sensitivity versus 1-specificity. RESULTS: The weighted k coefficient among observers was 0.46. The multivariative analysis did not identify any variable that could have caused such disagreement. Large differences in sensitivity and specificity were detected between observer number 1 (sens., 0.90, spec., 0.45) and observers number 2 (sens., 0.67, spec., 0.72) and number 3 (sens., 0.71, spec., 0.80), but the overall diagnostic performance (sensitivity vs 1-specificity) was superimposable in the three cases (p = ns). CONCLUSIONS: Simple, reproducible and agreed-on diagnostic criteria should be established to yield reliable results in a group of cytologists. The consideration of individual diagnostic performances can give a false idea of homogeneity between observers. In this field, concordance analysis makes quality control reliable and should be a routine procedure of any pathology department. PMID- 10464956 TI - [Indications from the laparoscopic approach in pediatric urological surgery: our experience in 49 patients]. AB - OBJECTIVES: To analyse the diagnostic and therapeutic potential of video-assisted laparoscopy in the urology paediatric pathology. We had analysed its indications and results, to measure in disminution of morbidity and hospital stay. PATIENTS AND METHODS: We are treated in our section of Paediatric Urology 49 patients during the period of June 1995 and December 1998, performing 51 laparoscopics procedures. The mean age was 8 years (2-16 years ago). The laparoscopic indications were diagnostic in 33 patients: intra-abdominal testes (n = 16), renal biopsy (n = 17), and terapeuthics in 16 patients (17 laparoscopics procedures): varicoceltomy (n = 7), orchiectomy (n = 1), closure peritoneal vaginal duct (n = 2), retroperitoneal nephrectomy (n = 6), marsupilazation and omentoplastic in giant lymphocel renal post-transplantation (n = 2). RESULTS: The laparoscopic approach were effective in 100% of diagnostic procedures and 93.4% of therapeuticm procedures. Conversion in 2 cases were necessary to make renal biopsy (n = 1). We hadn't have postoperatives complications and the indices of intraoperative morbidity was of 2.4%. The mean hospital stay was 1.4 days. CONCLUSIONS: We believe that exist indications clinically stabilised of laparoscopic approach--diagnostic and therapeutic--in paediatric urology. There are others indications that its are consider anecdotal in the present but in immediately future these indications will be valid. PMID- 10464957 TI - [Bladder transitional cell carcinoma rating, systematization of grading and value of DNA ploidy]. AB - TCCs are an heterogenous group of tumors with an uncertain biologic behaviour, especially intermediate grade (G2). Histologic grade and pathologic stage have revealed only partially useful on predicting the outcome. OBJECTIVE: Search and statement of objective and cuantitative parameters able to define prognostic subgroups to TCCs. MATERIAL AND METHOD: We studied consecutive 106 patients with TCC in order to: 1) Identify every variable with independent predictive value in classifying cases of TCC in a three (1,2,3) or two (high/low) grades systems. 2) Make a correlation between DNA ploidy obtained by image analysis and DNA ploidy and S-phase obtained by flow cytometry. RESULTS: Every variable used to define the histologic grade showed significative correlation with both gradation systems. In multivariate analysis, the presence of superficial cells and the mitotic counts revealed us the most valuable variables in predicting the histologic grade. DNA-ploidy (both obtained by static an flow cytometry) correlated well with grade, stage, growth pattern and necrosis, whereas S-phase did so with grade, mitotic index and DNA ploidy. Image cytometry showed similars results to those of flow cytometry and also was able to detect aneuploidy when an situ carcinoma or dysplasia were present in the adyacent mucosa. PMID- 10464958 TI - [Value of Markovian nuclear texture to predict progression in surface transitional cell carcinoma of the bladder]. AB - The aim of this work has been to determine if the DNA and nuclear Markovian textures of tissue sections evaluated by image cytometry correlate with the histologic grade and the progression probability of superficial transitional carcinoma of the bladder. In our study, DNA ploidy is related to histologic grade in that aneuploidy frequency increases with a higher grade of malignancy. Image analyses of superficial TCC tissue sections of the bladder, has allowed us to identify "sub-visual parameters" as well as nuclear textures that according to our results can be useful in clarifying the evolutive behaviour of these tumours. Two Markovian textures that identify entropy (TXI) and mean internal contrast (TXB) allow to discriminate between histologic grades as well as progression or non-progression. In conclusion, image analysis cytometry of paraffin embedded tissue sections of TCC of the bladder supplies densitometric parameters related to grade and provides valuable information for the prediction of progression. Quantification of chromatin pattern description in Feulgen-stained nuclei using the Markovian method can be useful in this context. PMID- 10464959 TI - [Relationship between preoperative urine cultures and prostatic gland cultures in patients treated for benign prostatic hyperplasia]. AB - OBJECTIVES: 1.--Investigate the bacteriuria preoperative in patients who will be operate on for Being Prostatic Hyperplasia (BPH). 2.--Define the prevalence of the prostatic colonization or infection. 3.--Try to correlate the bacteriological findings of urine and prostate, and find the degree of concordance between the microorganisms which can be commonly found in urine and prostatic tissue. METHOD AND MATERIALS: Prospective series of 175 patients undergoing prostatectomy for obstructive symptoms. The protocol revealed, among others variables: the preoperative urine culture; the presence or the absence of catheter; and the quantitative bacteriological culture of prostatic tissue. The information could be analised and its results could be obtained later on. The analysis stages consisted of both a descriptive and an analytic study. RESULTS AND CONCLUSIONS: 1.--Only 36 patients (20.6%) presented bacteriological increase of microorganisms (> or = 10(4) UFC/ml) in the preoperative urine culture. The Escherichia coli was the most common microorganism, followed by the Enterococcus faecalis, coagulase negative Staphylococcus and the Pseudomona aeruginosa. A single microorganism grew in 31 out of the 36 positive cultures. 2.--The prevalence of the infection or colonization of the prostatic tissue was 25.1% (44 patients). The most common isolated microorganism was the coagulase-negative Staphylococcus followed by the Escherichia coli and the Enterococcus faecalis, in concentrations of at least 10(4) UFC/gr of tissue in the 79.6%. A single microorganism was isolated in 32 out of 44 patients. 3.--The proportion of positive prostatic cultures, in patients with positive urine culture (38.3%), was significantly higher than the one obtained in patients with negative urine cultures (16.5%) (p < 0.0001). Nevertheless, 52.3% of the 44 patients with positive prostatic cultures had negative urine culture, and only 21 (58.3%) out of the 36 patients with positive urine cultures presented a bacteriological growth in prostate. The degree of concordance (Kappa index) between the microorganisms which were found in preoperative urine and prostatic tissue is low or none for the majority of them. PMID- 10464960 TI - [Changes in prostatic histological composition in subjects with prostatitis]. AB - OBJECTIVES: To explore the links between the histological composition of the prostate and the symptoms related to the clinical BPH. MATERIAL & METHODS: The prostates of 34 patients with symptoms of BPH and 17 asymptomatic patients were biopsied. Using light microscopy, the ratio of stromal to glandular surfaces was calculated. The main dimensions of 5 epithelial cells randomly selected, were examined under electron microscopy in each case. The number and length of microvilli, and the number of secretory granules were determined. The results of symptomatic and asymptomatic subjects were compared by means of Student's t and Mann-Whitney's U tests. RESULTS: The mean ratio of stromal and glandular surfaces came to 4:1 and 6.5:1 in the symptomatic and asymptomatic patients, respectively. The difference was not statistically significant (p = 0.18). Although the average cellular surface of the symptomatic patients was inferior to that of the asymptomatic subjects (74 and 88 mu2. respectively), the differences did not reach statistical significance (p = 0.14). Similarly, the mean nuclear surface of the symptomatic and asymptomatic patients (19.5 and 20.8 mu2, respectively) did not differ (p = 0.48). No differences were detected in the number of secretory granules and microvilli, nor in the length of the microvilli. CONCLUSIONS: The occurrence of symptoms related to the clinical BPH is not due to differences in the composition of the prostate. No clear links seem to exist between the structural changes in the epithelial cells and the development of symptoms. Considering the proportions of epithelium and stroma, therapy focused in the prostatic stroma should yield the best results. PMID- 10464961 TI - [Results of ambulatory major surgery in urology, within an integrated unit]. AB - OBJECTIVE: To contribute our experience in the performance of ambulatory major surgery, through the creation of an integrated unit. Such units take advantage of existing resources both in terms of space and materials, as well as personnel, without the need to open separate ambulatory major surgery units that would involve greater resources expenditure. MATERIAL AND METHODS: 48 patients undergoing ambulatory major urology surgery were analyzed. The patients were admitted the same morning of the procedure and then, after leaving the operating theatre, they were moved to the reanimation room and later to the urology ward, where they were evaluated by the urologist on duty and discharged. A few days later the patients returned to the outpatient's clinic for re-evaluation. RESULTS: 48 patients underwent surgery, 44 male and 4 female. Mean age of patients was 34 (3-86) years. The procedures most frequently performed were: hydrocelectomy, vaginal peritoneum duct closure, orchiopexy, ureterorenoscopy due to ureteral lithiasis, internal urethrotomy, varicocelectomy, vesical tumor resection and circumcision in children. Only in two cases, hospital stay was longer than 12 hours and there was only one re-admission. CONCLUSIONS: Ambulatory major surgery integrated units may allow to perform this type of surgery in small units, with little resources and with a volume of patients not sufficient to create an independent unit. PMID- 10464962 TI - [Kluyvera cryocrescens: a positive urine culture in a young girl with persistent proteinuria]. AB - Kluyvera genus usually shows two kinds of species: K. ascorbata and K. cryocrescens, DNA hybridization let us to differentiate a third group: Kluyvera species 3. Its diagnosis is quite uncommon and its taxonomy have been recently clarified. We report here a ten years female record with a chronic proteinuria and a positive urine-culture for K. cryocrescens. Axetil cefuroxime treatment was absolutely succesful. Kluyvera infections are difficult on the whole to joint with some specific clinical features. PMID- 10464963 TI - [Cystic dysplasia in the seminal vesicle. Case report and literature review]. AB - Contribution of one case report of cystic dysplasia of the seminal vesicle with ipsilateral renal agenesis in a 32 year-old male presenting with primary infertility. The seminograms showed moderate astenoteratospermia while in the abdominal-pelvic ultrasound there were changes in the right seminal vesicle and right renal agenesis, which was confirmed with further complementary tests: UIV, transrectal ultrasound, urethrocystoscopy, arteriography and renal scan. PMID- 10464964 TI - [Ectopic thoracic kidney]. AB - We present a case of kidney ecthopy in a 62 years old male patient, discovered by chance during complementary explorations realized in the diagnostic process of an acute respiratory pathology. We review the literature related to this anomaly: 200 cases have been published to the date. The thoracic kidney is a rare kind of kidney ecthopy, tipically without synomathology & it requires no treatment. PMID- 10464965 TI - [Painful erections related to sleeping]. AB - Case report of sleep-related painful erections in a 34 year-old male with grade C3 HIV infection. Due to severe impairment of the patient's general condition, no proper diagnostic studies were performed to gain deeper knowledge of the symptom's pathological etiology. Empirical therapy was started based on evidence from the literature consulted, and the results seen were optimal. This paper contributes a brief review of a condition infrequently seen by the vast majority of urologists. PMID- 10464966 TI - [Colonic perforation in percutaneous surgery of the kidney. Report of two cases]. AB - We show our experience about iatrogenic damage of colon secondarily to percutaneous surgery of the kidney. One of them treated by means of open surgery, the other by conservative procedures. Making discussion on risk factors, as well as the diagnostic therapeutical aspects. PMID- 10464967 TI - [Extragonadal germ tumor]. AB - Extragonadal primary tumors are uncommon and account for 3 to 5% of all germ cells neoplasias. Preferentially found in middle line structures, such tumors are frequently in an advanced stage when diagnosed. Differentiation between primary extragonadal tumors and metastasis of testicular primary tumors is essential, but may be difficult in many instances. PMID- 10464968 TI - [Paratesticular rhabdomyosarcoma]. AB - We report a case of paratesticular rhabdomyosarcoma in a six-year-old boy. The clinical presentation was subacute. The patient underwent a radical right inguinal orchidectomy. It was classified in IRS-III stage IA (based in the Third Intergroup Rhabdomyosarcoma Study). Subsequently, the child received 3 chemotherapy courses (9 weeks) with vincristine and actinomycin D. The patient is found to be asymptomatic 1 year after the treatment. PMID- 10464969 TI - [Contact-derived allergic balanoposthitis and paraphimosis through topical application of celandine juice]. PMID- 10464970 TI - [Gunshot wound to the genitals]. PMID- 10464971 TI - Adsorption studies of ciprofloxacin: evaluation of magnesium trisilicate, kaolin and starch as alternatives for the management of ciprofloxacin poisoning. AB - In vitro experiments were performed to investigated the extend of adsorption of ciprofloxacin to kaolin, magnesium trislilicate and to a starch obtained from the tubers of Tacca involucrata (Tacca starch) and to explore the effect of varying pH on this adsorption. Activated charcoal, a standard adsorbent and antidote in the management of poisoning due to a variety of chemical agents was employed as a comparing standard. The results of the study indicate that kaolin and magnesium trisilicate adsorbed ciprofloxacin effectively while the adsorption of the drug on the starch was relatively low. Adsorption was dependent upon the quantity of the adsorbed used. Kaolin or magnesium trisilicate could serve as an effective antidotal alternative to activated charcoal in the management of ciprofloxacin poisoning. Except in cases of poisoning due to ciprofloxacin, the concurrent administration of the drug with kaolin or magnesium trisilicate may be contraindicated. Tacca starch, however, may not really be recommended for the management of ciprofloxacin poisoning. PMID- 10464972 TI - Study of accelerated storage conditions affecting physical characteristics, in vitro dissolution and stability of bioadhesive containing tablets AB - The effect of storage at different temperatures and relative humidities on directly compressed tablets of indometacin containing 20% polycarbophil was studied for up to 16 weeks. The prepared tablets were stored in closed containers of type III glass under different conditions of temperature (30 degrees C, 40 degrees C and 50 degrees C), in open petri dishes at room temperature, at relative humidity (RH) (31%, 79.3% and 98%), and combinations of temperature and relative humidity (30 degrees C/92.9% RH, 40 degrees C/79.5% RH and 50 degrees C/65% RH). The tablet properties such as mean weight, hardness, disintegration, in-vitro dissolution and drug content were monitored just after preparation and after 2, 4, 8, 12 and 16 weeks of storage. The tablets mean weight was significantly (p < 0.05) increased at RH 98% and at 30 degrees C/92.9% RH and non significantly increased at 79.3% RH, 40 degrees C/79.5% RH and 50 degrees C/65% RH. There was no effect of temperature on mean weight. The hardness of the tablets increased slightly at 31% RH and decreased at 79.3% RH and at 40 degrees C/79.5% RH and became zero at 98% RH and at 30 degrees C/92.9% RH. The increase in temperature caused slight increase in tablets hardness. Tablets stored at 98% RH and at 30 degrees C/92% RH showed an increase in disintegration time associated with a decrease in rate of dissolution and indometacin content. No effect on drug content or dissolution profile was observed as a result of storage under other conditions. These findings point out to the importance of proper protection of indometacin tablets containing polycarbophil from high temperature and humidity conditions in order to ensure stability of the drug. PMID- 10464973 TI - Determination of ametryn herbicide by bioassay and gas chromatography-mass spectrometry in analysis of residues in drinking water. AB - A simple, rapid and quantitative bioassay method was compared to a gas chromatography/mass spectrometry (GC/MS) procedure for the analysis of ametryn in surface and groundwater. This method was based on the activity of ametryn in inhibiting the growth of the primary root and shoot of germinating letuce, Lactuca sativa L. seed. The procedure was sensitive to 0.01 microgram/l and was applicable from this concentration up to 0.6 microgram/l. Initial surface sterilization of the seed, selection of pregerminated seed of certain root lengths and special equipment are not necessary. So, we concluded that the sensitivity of the bioassay method is compatible with the chromatographic method (GC-MS). However, the study of the correlation between methods suggests that the bioassay should be used only as a screening technique for the evaluation of ametryn residues in water. PMID- 10464974 TI - Thermodynamic ionization constants of antihypertensive drugs. AB - The thermodynamic ionization constants (pKa) of indapamide, clonidine and methyldopa have been determined at 25 degrees C and 35 degrees C by pH titration technique. The pKa varies linearly with mole fraction of dioxane. The thermodynamic parameters have been calculated. PMID- 10464975 TI - Synthesis of pharmacologically active indoles. AB - Formylation of 6-methoxy-1-methyl and 5-methyl,2,3-diphenyl-1H-indole (Ib and IX) gave the 5- and 6- carboxaldehyde derivatives (II and X) respectively, which were treated with ethyl cyanoacetate to form the corresponding 2-cyano-3-substituted acrylic acid ethyl ester (III and XI). The latter compounds reacted with hydrazine hydrate, urea and thiourea to form the corresponding 5-amino-4 substituted 2,4,dihydropyrazol-3- one (IV), 6-indolyl-2,4-dioxo-1,2,3,4 tetrahydropyrimidine-5-carbonitrile s (V and XII) and 6-indolyl-4-oxo-2-thixo 1,2,3,4-tetrahydropyrimidine-5-ca rbonitriles (VI and XIII). Reaction of the 5- and 6-carboxaldehyde derivatives with malononitrile afforded the 2-substituted malononitrile derivatives (VII and XIV). VII and XIV reacted readilly with aromatic ketones to give the 2-amino4,6-disubstituted nicotinonitriles (VIII a,b and XVa,b). The biological activity of compounds Ia, Ib, II, III, IX and X was tested for antiinflammatory, ulcerogenic and antispasmodic activities. PMID- 10464976 TI - Cytotoxic activity against a series of tumour cell lines of dimethyltin dichloride complexes with various donor ligands. AB - Several dimethyltin dichloride complexes of the general formula Me2SnCl2.L, where L = 8-hydroxy quinoline (L1), 8-hydroxy quinoline-N-oxide (L2),2,6 diaminopyridine (L3), 3-amino aniline (L4), cyclohexyl amine (L5), (1R, 2R)-1,2 cyclohexane diamine (L6), 3,5-dimethylpyrazole (L7), harmaline (L8) and harmine (L9), have been prepared and characterized physicochemically and spectroscopically. It was found that tin coordinated with the active site of these ligands to give complexes with penta-coordinated tin or, to some extent, hexa-coordinated tin. The cytotoxic activities of these complexes have been studied against Hep-2, HeLa, L20B and RD lines using the MTT-colorimetric assay. These activities were compared with those of the three reference standards, cisplatin, carboplatin and oxaliplatin. Some of these complexes exhibited a range of significant activities against the cell lines used, whereas no cytotoxic activities were exhibited by the rest of these complexes. The significance of these results is given and discussed. PMID- 10464977 TI - Anisylidine pyruvic acid in heterocyclic synthesis AB - A series of substituted pyran, pyridine, thiin (3), (4), (5) and (6) were prepared from anisylidine pyruvic acid (1a). The indenopyran (7) was also prepared from (1a) via Michael addition reaction. Compound (1a) condensed with o phenylenediamine and o-aminophenol to give (8) and (9). Reaction of (1a) with thiosemicarbazide afforded (10) which cyclized to give the triazine (11). Compound (1b) was subjected to react with o-phenylenediamine, o-aminophenol and diamino-arylazopyrazole derivatives to yield the Schiff bases (12, 13) and (14) respectively. Addition of malononitrile (1 mole) to (1b) gave (15) while addition of (2 moles) of malononitrile gave (16). Reaction of (1b) with urea yielded the pyrrolopyrimidine (17). The structures of the products were confirmed by their correct analytical data. PMID- 10464978 TI - Synthesis and in vitro antifungal activity of some N,N-disubstituted dithiocarbamic acid esters derived from 2-methylquinazolinones. AB - A series of 2-[(N,N-disubstituted thiocarbamoylthio)methyl]quinazolinones 9a-g; 10a; 10d; 11a-d and 12a were synthesized and evaluated for potential antifungal activity against a variety of fungal species. The synthesis of the target compounds was achieved by reaction of the potassium salts of disubstituted dithiocarbamic acids 8a-g and the respective 2-bromomethyl-4(3H)-quinazolinone 4 or 3-aryl-2-chloromethyl-4(3H)-quinazolinones 5-7. The dithiocarbamic acid derivatives were synthesized in a one step reaction from the appropriate amine, alcoholic potassium hydroxide solution and carbon disulfide. TLC and elemental analyses ascertained the purity of the synthesized compounds and their structures were confirmed by IR and 1H-NMR spectroscopy. 2-Methyl-4(3H)-quinazolinone 2, the precursor of the 2-bromomethyl intermediate 4, was selected as representative example for detailed spectroscopic investigations, including 300 MHz 1H- and 13C NMR in addition to HH COSY; APT and 1H13C HETCOR spectra, with the aim of establishing correct assignment of the spectral data of related compounds. The synthesized disubstituted dithiocarbamates 9a-g; 10a,d; 11a-d and 12a as well as tolnaftate and clotrimazole, as reference drugs, were tested in vitro at 2 and 5% concentrations against 23 pathogenic fungi. The study revealed that compound 9a exhibited broad spectrum inhibitory activity that is superior or comparable to that of the reference drugs against the tested fungal isolates. Selective fungistatic activity against Candida species was elicited by compound 9e and against Microsporum species as well as Trichophyton mentagrophytes was also observed for compound 9g. As a general pattern it might be postulated that some of the synthesized dithiocarbamate derivatives showed broad spectrum antifungal activity as compared with tolnaftate, the clinically used thiocarbamate compound, and also exhibited comparable activity to clotrimazole against Candida species and F. Solani. PMID- 10464979 TI - Training the trainers. PMID- 10464980 TI - Link between tooth loss and radiation. PMID- 10464981 TI - Graduate concerns. PMID- 10464983 TI - Anxiety, phobias and sedation. PMID- 10464982 TI - Dentists obliged to consider sedation. PMID- 10464984 TI - Serious implications to sedation debate. PMID- 10464985 TI - Conventional crown and bridgework. AB - The restoration of worn teeth with crowns requires a number of strategies to create space for retentive, resistant and durable castings. PMID- 10464986 TI - The emergency drugs box--time for action? AB - Many GDPs are confused about which emergency drugs they should stock. There is a lack of uniform advice on this topic. Most recommended lists of emergency drugs contain superfluous medications which GDPs would never be expected to use. This paper rationalises the content of the emergency drugs box and provides clear, standardised guidance. A list of ten essential emergency drugs is presented, all of which can be administered by simple routes. Every GDP should be trained and competent to use these essential emergency drugs. PMID- 10464987 TI - Barriers to accessing and accepting dental care. AB - This paper looks at the psycho-social factors influencing patients use of dental care, and what the dentist can do to help when things go wrong. PMID- 10464988 TI - A 3 to 4 year study of single tooth hydroxylapatite coated endosseous dental implants. AB - OBJECTIVE: To evaluate the clinical effectiveness, common complications and maintenance associated with hydroxylapatite (HA) coated cylindrical implants when used to support single crowns. DESIGN: A prospective medium term clinical study of the Calcitek HA-coated implant. SETTING: Implant placement, crown fabrication and follow-up procedures were carried out at the Leeds Dental Institute, between 1990 and 1998. SUBJECTS AND METHOD: 26 patients (33 implants) participated in the trial. They were referred from general dental practitioners because of their suitability for single tooth implant placement. MAIN OUTCOME MEASURES: The implants were assessed using recognised clinical review procedures e.g. radiographs and soft tissue assessments. RESULTS: At exposure there was 100% implant integration. The cumulative survival rate over 4 years was 100%. In five implants there was cervical bone loss of more than 4 mm and these were classified as failing. This gave an overall cumulative success rate of 58% by year 4. CONCLUSION: The Calcitek HA-coated single tooth implant shows exceptionally high initial integration however, the longer term results suggest that the cervical bone level adjacent to the implant failed to establish a steady state. Doubts remain regarding the long-term prognosis of these cylindrical HA-coated implants. PMID- 10464989 TI - The dental caries status of Scottish adolescents reported to be regular attenders. Initial results from a primary dental care based research network. AB - AIM: To investigate the caries status of, and the delivery of care to, a group of regularly attending adolescent dental patients. To conduct research in primary dental care. A subsidiary aim was to compare the caries status of this sample to population samples. SETTING: General dental practices across Scotland. DESIGN: A 3-year cohort study. SUBJECTS: 41 volunteer general dental practitioners and 616 adolescent patients (mean age = 12.1 years at baseline) defined by the practitioners as 'regular' attenders. RESULTS: All practices remained in the study. 403 subjects were seen at both baseline and final examination and 329 were examined at all 4 annual examinations. The mean D3MFT (dentine caries threshold) was 1.8 at baseline and 3.9 at the final examination, three years later. Of the 541 subjects seen at baseline 62% had experienced either restored or unrestored dentinal caries. Thirty-four percent of all those examined at baseline had all the unrestored dentinal caries. The majority of the disease was to be found in the molar teeth, particularly the first permanent molars. The provision of sealants was relatively high with 85% of subjects having at least one sealed tooth by the final examination. Although disease levels were related to socio economic status, the Care Index was not. CONCLUSIONS: The 'regularly attending' subjects had a better normative level of dental health than their peers in the Scottish population. However, wide variation was found. The project also demonstrated the feasibility of undertaking research in partnership with general dental practitioners. PMID- 10464990 TI - Using problem-based learning as part of general dental practice vocational training. AB - The use of problem-based learning as part of a vocational training scheme for vocational dental practitioners is described and evaluated. Problem-based learning is currently viewed as an effective means of delivering medical education and has been adopted as a principal style of education by many undergraduate medical and dental schools around the world. PMID- 10464991 TI - [Prescriptions and consumption of venotonic drugs in France (a propos of the report of the French National Institute for prescriptions and consumption of drugs)]. AB - In 1997, consumption of veinotonic drugs in France represented 3.8% of the sales of refundable patient medicines, that is to say nearly 3 Billion francs with a 2.8% mean annual increase in units sold from 1983 to 1997, whereas prices decreased and new presentations with stronger dosages and larger packagings appeared on the market. No French epidemiologic specificity could be evidenced as regards the prevalence of chronic venous insufficiency and its complications. In France, and in comparison with elastic support, veinotonic products are preponderant in the treatment of chronic venous insufficiency despite restrictive labelling of authorization (treatment of symptoms) and recommendations of good practice which consider them as a complementary treatment. They are the subject of 18 million prescriptions per year, by 69,000 doctors, mainly general practitioners. They are utilized over long periods, within a context of prescription rather than automedication. The number of patent medicines available (109), the status of refundable medicine, strong demand of patients and therapeutic habits have made France the world's foremost market for veinotonic products, with 70% of international sales. French particularism is indeed due to these factors and not to the prevalence of the condition to be treated. PMID- 10464992 TI - [Palliative care, a medical discipline dealing with the end of life]. AB - Insofar as they take into account, not only the physical needs of the dying, but also their psycho-affective ones, Palliative Care are, with time, becoming a full fledged area of medical specialization, the one which deals with the last phase of life. In order to be really efficient, that medical activity requires to resort to numerous skills: those of doctors, nurses, psychologists, physiotherapists..., which implies setting up suitable structures. Historically, Palliative Care have developed within specialized units: the Palliative Care Units; such units should, now, only be seen as centres for research and teaching. But, as time goes one, Palliative Care should, in fact, be practised wherever patients are dying: i.e., in hospitals, provided the staff are adequately trained and even, if need be, with the help of Mobile Palliative Care Teams; at home where general practioners, helped by a real network of professionals, could then manage to give the patients who wish to do so, the opportunity to die at home. Such practise of Palliative Care makes attitudes of therapeutic obstinacy irrelevant. Moreover when Palliative Care are practised, patients practically never ask for euthanasia. PMID- 10464995 TI - [Is it necessary to legislate euthanasia?]. AB - There are no specific articles on the end of life in French law. Thus an act of euthanasia can be qualified as murder, murder with premeditation or non assistance to a person in danger. Recent events and debates have raised the question of enacting new legislation to deal with this problem. Two contrary positions could be considered: either create a special offence or explicitly authorize acts of euthanasia. There are major objections to both these propositions. The first one would require taking account of various situations, - unbearable suffering, loss of dignity, and precise requests,--that would be impossible to specify in legislative terms. The second proposition would be open to the same objection; it would also derogate from the major principle of respect for the life of persons and thus risk setting a precedent that could be used in other circumstances to evade this principle. Finally, if a law were enacted to this effect, it would constitute a threat for the development of palliative care. PMID- 10464994 TI - [Is there any deontology of accompanying people at the end of life?]. AB - The thought on ethics, enhances by accompanying patients or human-beings reaching end of life, belong to major themes which have for years been dealt with by philosophers and monks. The physicians, who by nature accompany and live with their patient this life period a sometimes close relationship, have conceived within deontology a wonderful expression of what ethics inspired them. Reading articles 37 and 38 of the Code of Ethics brings a clear, precise and qualified answer to the question raised by the title. We easily find out the main principles which would be the guideline to the doctor's behaviour coping with this situation: providing suitable and continue cares to maintain the quality of a life which comes to end, respecting dignity. Relieving suffers, morally assisting, comforting the family, knowing to limit the cares reasonably. The sensitive and human expression sums up pretty well the particularities essential to all (physicians, health professionals or volunteers) who have accepted this heavy task. PMID- 10464993 TI - [Treatment of different types of pain]. AB - Accurate assessment of pain is the key to appropriate analgesia. This necessitates not only an understanding of the organic component, but also a comprehension of the role played by the mental, social and spiritual dimensions in the individual patient's suffering. It implies that the entire care team must be involved in pain management. The nature of a patient's pain is one predictor of the response to treatment. It is mainly characterized by its location, intensity, extent, timing and type (excessive nociception, neurogenic, or mixed), the circumstances in which it appears, and any accompanying signs. The choice of analgesic for treating pain due to excessive nociception was greatly facilitated by the introduction of the WHO three-step approach. Better knowledge of the pharmacological and pharmacokinetic properties of the different analgesics has contributed to increase their efficacy and tolerability. Certain types of pain of neurogenic origin respond poorly to both opiate and non opiate analgesics. They can be treated with other drugs whose mechanisms of action in pain relief are not fully understood. They include the following; antidepressants (amitriptyline, nortriptyline, desipramine and doxepine); anticonvulsants (carbamazepine, phenytoin, valporic acid and clonazepam); antiarrhythmic agents (lidocaine, mexiletine, flecainide and tocainide). The unwanted effects of these different treatments, together with psychological disturbances induced by the underlying disease, can call for the use of antiemetics, laxatives, spasmolytics, glucocorticoids and psychotropic agents (anxiolytics, neuroleptics and antidepressants). Finally, in many cases, better pain relief is obtained by combining drug-based therapy with other interventions such as radiation therapy, neurosurgery, and psychological/behavioral approaches. All these means must be chosen and used according to each individual patient's needs. Pain must be considered as a disease that can and must be eliminated or at least attenuated. PMID- 10464996 TI - [Is it possible to die a good death?]. AB - Is there any sense in wondering if one can "die a good death" Indeed, the word "euthanasia" as such offers food for thought. Obviously, human beings must die. The only real question is: Can help and assistance be given (through others) to a person when facing this ineluctable issue; and, if so, in which way and how far? To make things clearer, two points may be taken into consideration. On the one hand, it is a fact that the roman-catholic tradition has been able to set up a practice of a "good death piety", or even a "good death pastoral". But, on the other hand, we must not forget that when we consider what "euthanasia" means for the end of life and bear in mind what "eugenism" means for the beginning of life as well as the way it has been used, then it can only put us on the alert for possible serious abuse.... Considering all this, the question may be put as follows: not: is it possible, under certain circumstances, to give someone his or her death; but: how must I help and assist a person to live his or her life to the very end, if possible up to death? PMID- 10464997 TI - [Spiritual needs at the end of life]. AB - All human beings have been dieting since two million of years, and since 100,000 years they have surrounded death with rites and religious considerations, which reflect a basic need to assume the end of life with spirituality. Religion helps to prepare patients spiritually in the end of their life to cope with death as something which cannot be eluded. We are all bound to die. A spiritual fullness should contribute to the serenity and to strengthening of the soul at the end of life. PMID- 10464998 TI - [To go along with life until death]. AB - We are faced with difficult and complex questions that cannot be answered by stating great principles or ideological convictions, because they refer to painful situations, always singular, in which each individual, in a unique way, faces his life and his death. But the debate can draw on shared convictions and values. Thus, before being a way of assuming death, the Christian faith is fundamentally a way of welcoming the life, in all times and in its fullness, that Christ has given us. The whole Bible and in particular the ministry of Jesus bear witness to that fight for life, against the scandal of suffering and the powers of death that are a denial of the good work of God. Suffering is never, as such, acceptable or justifiable, and truth is never to surrender to it, as if it was a meaningful destiny. And so all suffering that can be avoided must be so. Regarding death, it is often held back in the margins of our lives and societies, as if it was a sort of setback for our human abilities and especially for medicine. Of course those abilities exist but death is not an illness. It's the natural mark of our human finiteness and there is a time when caring is not intended to cure, but to make up for life that defaults, alleviate suffering. That is why what is called to-day palliative caring is so important. Because even when medicine is powerless in front of illness, it can still do something for the sick. Because of all that, the believer can only be opposed to euthanasia which is, after all, only the exact replica of the useless prolongation of life by medical means it pretends to oppose. It's the same activism, the same pretense, the Bible fights, through which human beings want to remain the masters of life and death. But death is not given, except in deathly violence. As life, it is welcomed and is accompanied. The end of a life is still life. To die is to the live to one's last breath. And that questions the claim to die "with dignity" when life can no more be lived in a "dignified way". But what "dignity" are we talking about? To-day, do we not mistake it with the image of the modern individual, master of himself and a match for the world, assured of his physical strength and of his conscientiousness enabling him to consent. As if the image of dignity was always the same, at every age, for all types of illness or simply of existence. When that image of dignity gets shaky, one discovers sorts of dignity that do not answer those criteria, but that testify that any "body" can be a subject, and that we know nothing about it. Lastly, no law or moral authority, be they lax or restrictive, can suppress compassion, nor the ethical responsibility of the patient, of the doctors, of the family circle. None can take the place of the common requirement: go along with life up to death. PMID- 10464999 TI - [Life from beginning to end]. AB - Each instant of life is essential, without us being able to distinguish which is the first and which is the last, without letting go the least of instant, under the cover of our questions concerning death. We have to notice the emergency of experts who take in charge the time following death, when it was precedently the task of the clergy and the relatives. When the doctors, having done everything in their power to save the life of a patient are confronted with the obligation to lessen the level of the medications, particularly for the more complex cases, they only bring back men in his status of men, faced with his death. But to spare his sufferings is an imperative of a humanitarian gesture. The procedure of the establishment of a law in the Senate, slips under the legislation of active euthanasia, which is the open door to every thing we had always refused. Helping someone who is suffering is a given, but killing him by contributing by any mean which would shorten his life, is an important fault which repercussions could shape the future of society at large. The emphasis on the particularity of each case, has to be stressed and it is in this aspect of the unique, that human reflexion will be preserved. A law could be dangerous in the sense that it might "normalize" what should never be. The doctors by taking away some of the pain, even if it is by administrating prescriptions which could cause death, are not doing any thing but holding on to their engagement of taking care of the patient, which implies and consists in not harming him. PMID- 10465000 TI - [Estrogen-induced genes in breast cancer, and their medical importance]. AB - The study of several human estrogen receptor positive breast cancer cell lines has allowed characterization of a number of estrogen-induced proteins (e.g. progesterone receptor, cathepsin D, pS2 and fibulin-1 in ovarian cell lines). In primary tumours, these markers have different prognostic significance for predicting whether the tumour will be hormone responsive (e.g. pS2, estrogen and progesterone receptors) or will develop metastasis (e.g. cathepsin D). Studies of estrogen-regulated genes should also lead to new therapeutic approaches for hormone-resistant cancers. The role of estrogens as mitogens stimulating the growth of breast and ovarian cancer cell lines is well established. By contrast, their action on metastasis appears more ambiguous. Breast cancer cells without estrogen receptor (ER) are generally less differentiated and more aggressive than those containing functional ER. Moreover, the reexpression of ER by transfection in ER-negative cell lines inhibit their metastatic and invasive potential. These results suggest a protective role of ER in tumor progression. Studies of the underlying mechanisms of this effect may open new therapeutical strategies. PMID- 10465001 TI - [Is it possible to reduce the incidence of aminoglycoside-induced nephrotoxicity?]. AB - The incidence of nephrotoxicity due to aminoglycosides should be sharply reduced. The indications for prescribing these antibiotics should be limited to infectious disorders induced by aerobic Gram-negative bacteria and by some Gram-positive bacteria requiring treatment in specialized hospital units using an association of aminoglycosides and another antibiotic. Daily doses should not exceed those indicated by the manufacturer, and the length of treatment should be as short as possible, with a relay to other antibiotics that are not or are less nephrotoxic. The possibilities for reducing the incidence of nephrotoxicity are few. It is not possible to prevent the antibiotic from entering the renal tubular cell or from producing deleterious effects therein. However, by using short-term intravenous infusion as the administration route, prolonged contact between the antibiotic and its receptors on the brush borders of the proximal tubular cells can be avoided, particularly since the process of cellular absorption is saturable. Essentially, doses should be adapted according to the age and the glomerular filtration of the patient, since renal function usually decreases with age. Volemic and hydroelectrolytic disorders favour nephrotoxicity and should be corrected. Associations with other nephrotoxic drugs should either be avoided or used with increased caution. The same is true in special situations such as endotoxaemia, severe renal parenchymatous infections and cholestasis. In any case, given the well-known insidious onset of nephropathy, aminoglycoside treatment always requires laboratory follow-up consisting of repeated testing of creatinemia during the two weeks of treatment. PMID- 10465002 TI - [Thromboembolic syndrome from prolonged sitting and flights of long duration: experience of the Emergency Medical Service of the Paris Airports]. AB - The thromboembolic syndrome due to prolonged sitting was described after varied circumstances, particularly among the passengers after a long duration flight. The Aeroports De Paris medical department had collected 70 cases of pulmonary embolism between 1984 and 1998. Their incidence increases during the last years, corresponding to the growth of air traffic and mainly to the increase of long duration without stop flight (76.5% of our cases correspond to more than 12 hours flight). According to the number of the passengers landing in the Aeroports de Paris, the incidence during 1998 is 0.5 per million of passengers, with an important female prevalence and 31% of cases with thrombo-embolic antecedents. 61% of reported cases fainted just after landing. Our experience is limited as it considers only the acute cases, the late accidents do not come in our emergency service. PMID- 10465003 TI - Vaccination programme for group C meningococcal infection is launched. PMID- 10465004 TI - Double labelling to obtain S phase subpopulations: application to determine cell kinetics of diploid cells in an aneuploid tumour. AB - We studied the cell kinetics of the murine mammary carcinoma MCa-K using iododeoxyuridine (IdUrd) and chlorodeoxyuridine (CldUrd) given at different times as independently detectable labels of S phase cells. The presence of IdUrd and CldUrd, and the amount of DNA were measured by three-colour flow cytometry making it possible to define three subpopulations within S phase and to measure the progression through the cell cycle during the time following labelling. In DNA histograms of these subpopulations, the diploid and aneuploid cells (which had a DNA index of 1.7) are essentially completely separated. From appropriate combinations of cells labelled with IdUrd only, CldUrd only, or both, it was possible to construct separate DNA distributions for the labelled diploid and aneuploid cells at the times of administration of each label. The kinetics of the diploid and aneuploid cells could be calculated for individual tumours from these two time points without having to make corrections for the presence of the second population. The diploid and aneuploid populations had indistinguishable S and G2 + M phase durations, T(S) and T(G2 + M), of about 9 and 2 h; however, the potential doubling time values for the aneuploid and diploid populations were 30.2 and 101.2 h respectively. PMID- 10465005 TI - Regulation of differentiation and protein kinase C expression in 3T3 T proadipocytes: effects of TGF-beta and transformation. AB - We are studying the mechanisms that regulate proliferation and differentiation of normal 3T3 T proadipocytes and neoplastically transformed clones which have lost the ability to differentiate. The phorbol ester 12-O-tetradecanoylphorbol-13 acetate (TPA) and transforming growth factor beta (TGF-beta) are known inhibitors of the same step of the differentiation process in 3T3 T cells. Here, we examined the expression of the phorbol ester receptor/protein kinase C (PKC) during adipocytic differentiation of 3T3 T cells and its modulation by the differentiation inhibitor TGF-beta. PKC receptor assays were performed using a tritiated analogue of TPA and it was found that PKC receptor levels decreased approximately threefold during differentiation. Northern blot analyses revealed an even greater decrease of PKC transcripts during differentiation. TGF-beta inhibited not only differentiation, but the differentiation-dependent decrease in PKC levels as well. Transformed 3T3 T cells which have lost the ability to differentiate were found to express aberrant levels of PKC. The data suggest that TGF-beta may inhibit differentiation via a PKC-dependent pathway and that disruption of normal PKC levels or its regulation may be involved in the loss of differentiation control in transformed 3T3 T cells. PMID- 10465006 TI - Effects of mitomycin C and porfiromycin on exponentially growing and plateau phase cultures. AB - Laboratory studies and clinical trials are exploring the use of hypoxia-directed cytotoxic agents as adjuncts to radiotherapy. Because hypoxia and the microenvironmental inadequacies associated with hypoxia in solid tumours inhibit cell proliferation, an essential requirement for the successful use of hypoxia directed drugs in cancer therapy is that these drugs be toxic to quiescent tumour cells, as well as tumour cells progressing rapidly through the cell cycle. The experiments reported here compared the cytotoxicities of mitomycin C and porfiromycin to exponentially growing and plateau phase cultures of EMT6 mouse mammary tumour cells. The proliferative status of the cultures did not influence the cytotoxicity of mitomycin C under either aerobic or hypoxic conditions, or the cytotoxicity of porfiromycin in air. Exponentially growing cultures were slightly more sensitive than plateau phase cultures to porfiromycin in hypoxia, but the difference between the sensitivities of proliferating and quiescent cells was much smaller than the difference between aerobic and hypoxic cells. No evidence for repair of potentially lethal damage was found after treatment with porfiromycin in air or in hypoxia; this is in agreement with previous findings for mitomycin C. Mitomycin C and porfiromycin therefore exhibit the toxicity to quiescent cells needed for effective use as hypoxia-directed drugs for the treatment of solid tumours. PMID- 10465007 TI - FB1, a monoclonal antibody reacting with a keratin 14 epitope, stains only a small subset of psoriatic basal keratinocytes. AB - A murine monoclonal antibody, FB1, reacted with the basal keratinocytes of human stratified epithelia. One-dimensional and two-dimensional immunoblotting assays, performed on keratins extracted from HaCat cells and normal human keratinocytes, showed that FB1 recognizes K14. When LL002, another K14 monoclonal antibody is added, the FB1 stained area in the 2D-immunoblot seems to cover a fraction of the LL002 spot. Immunohistochemical data obtained from studies on normal human tissues supported the K14 specificity of FB1, but when compared with two other monoclonal antibodies, LL002 and RCK107 reacting with K14, some differences appeared. These differences were mainly seen in sweat glands, hair follicles, psoriatic epidermis and salivary glands. In psoriatic epidermis, FB1 showed a heterogeneous pattern of staining of the basal cell compartment. Intense reactivity was only observed at the bottom of the rete ridges. Staining diminished and finally disappeared in the basal cells above the dermal papillae. This observation supports the view that an increased germinative cell population in psoriasis involves a partially differentiated amplifying compartment in which the number of cell divisions is increased. PMID- 10465008 TI - The proliferation-associated Ki-67 protein: definition in molecular terms. PMID- 10465009 TI - Abnormal DNA synthesis in polyamine deficient cells revealed by bromodeoxyuridine flow cytometry technique. AB - Chinese hamster ovary cells were seeded in the absence or presence of the polyamine synthesis inhibitor 2-difluoromethylornithine (DFMO). At 1-4 days after seeding, the cells were labelled for 15-120 min with the thymidine analogue bromodeoxyuridine (BrdUrd) and they were then fixed directly after the labelling period. In addition, cells were labelled for 30 min and they were then allowed to progress in BrdUrd-free medium during a defined post-labelling time before fixation. An indirect immunofluorescence technique, using the monoclonal BrdUrd antibody and the intercalating stoichiometric DNA stain, propidium iodide, was applied to enable quantification of cellular BrdUrd and DNA contents, respectively, by flow cytometry (FCM). By comparing the mean DNA content of BrdUrd-labelled cells to the mean DNA contents of G1 and G2 cells, a relative measure of the position of the BrdUrd-labelled cells was obtained (relative movement). Relative movement data, obtained from control and DFMO-treated cells fixed directly after BrdUrd labelling, indicated that DFMO-treated cells entered S phase at a normal rate, while their progression through S phase was impaired. DNA histograms of BrdUrd-labelled control cells fixed directly after labelling showed that most cells were found in early and late S phase, while DNA histograms of BrdUrd-labelled DFMO-treated cells showed that most cells were in early S phase, indicating a delayed progression through S phase. Analysis of relative movement of cells that were allowed to progress in BrdUrd-free medium after labelling showed that DFMO treatment resulted in a significant lengthening of the DNA synthesis time. Labelling index was significantly higher in DFMO-treated, growth-inhibited cells than in early plateau phase control cells indicating an S phase accumulation in the former cells. PMID- 10465010 TI - Expression of the 170-kDa and 180-kDa isoforms of DNA topoisomerase II in resting and proliferating human lymphocytes. AB - The expression of the 170-kDa (alpha) and the 180-kDa (beta) isoforms of DNA topoisomerase II (topo II) was investigated with two specific monoclonal antibodies (MoAbs) in human peripheral blood lymphocytes (PBL), before and after phytohaemoagglutinin (PHA) stimulation. Binding of each MoAb was detected by indirect immunofluorescence labelling and quantified with flow cytometry. In resting PBL, the intensity of immunostaining was very low for both isozymes; however, topo IIbeta-associated immunofluorescence was about 2.5 times significantly higher (P<0.001) than that associated with the alpha isoform. Between 48 and 72 h of PHA stimulation, when the highest percentage of cells in S and G2+M phases was found, the levels of topo IIalpha and beta increased up to about 30 and 10 times the value measured in resting PBL, respectively. Thus, the two isoforms reached comparable immunofluorescence values. At longer stimulation periods (96-120 h), topo IIalpha immunofluorescence was not significantly changed, while that relative to topo IIbeta declined to about 50% of the peak value (P<0.02). At this time however, topo IIalpha-associated immunofluorescence was not significantly different from that related to the beta isozyme. These results suggest that in resting PBL topo IIalpha is required at levels lower than topo IIbeta, while in proliferating lymphocytes both isoforms are expressed to significantly higher levels. PMID- 10465011 TI - Expression of myc, fos and Ha-ras associated with chemically induced cell proliferation in the rat liver. AB - Events secondary to induced cell proliferation may play a role in the carcinogenic process. These studies investigated the expression of genes associated with growth control in response to two types of cell proliferation stimuli in the livers of male F344 rats. Regenerative hepatocyte proliferation after partial hepatectomy or a single dose of carbon tetrachloride, and mitogenic liver hyperplasia induced by a single dose of phenobarbital or WY-14,643 were assessed by thymidine incorporation and quantitative autoradiography. The expression of myc, fos, and Ha-ras was evaluated by Northern blot analysis of liver derived poly(A)+ mRNA from these same animals. After each treatment, the level of hepatocyte proliferation (labelling index 4-32%) was observed to peak between 24 and 48 h and return to control values by 8 days. In every case, a peak in myc expression was seen between 0.5 and 18 h depending on the proliferative stimulus treatment. A large peak in fos expression was seen at 0.5-2 h but only with the cytotoxic and regenerative proliferative treatments partial hepatectomy or carbon tetrachloride. A broad peak in Ha-ras expression was observed 12 to 36 h after each treatment. These data demonstrate transient expression of these genes following the synchronous induction of hepatocyte proliferation. The increased expression of fos upon treatment with cytotoxicants, but not mitogens, suggests different modes of growth regulation that may be important in understanding the induction of cell proliferation by these two types of agents. PMID- 10465012 TI - Slowly cycling (label-retaining) epidermal cells behave like clonogenic stem cells in vitro. AB - Slowly cycling label-retaining epidermal cells were identified by light microscopic autoradiography in the dorsal epidermis and hair follicles of adult mice 8-10 weeks after twice daily injection of [3H]dT on days three through five after birth. Pulse-labelled epidermal cells were identified in the epidermis and hair follicles of 7-8 week old mice 1 h after a single injection of [3H]dT at 8.00 a.m. For mice of both groups, epidermal cells including those from the hair follicles were harvested by trypsinization and were cultured from low density on feeder layers of irradiated Swiss mouse 3T3. On days 2, 4, 5, 7, 10 and 12, the cultures were fixed and processed for light microscopic autoradiography, and the distribution of labelled nuclei was quantified. On day 2 of culture, both label retaining cells (LRC) and pulse labelled cells (PLC) were found primarily as single cells. After five days, LRC were found as pairs and clusters having silver grain counts consistent with their division. In contrast, PLC remained primarily as single cells. These results suggest that LRC may divide to form colonies (are clonogenic) whereas PLC are rarely clonogenic. The significance of this experiment is that it suggests that the LRC may not only be persistent in the epidermis, but that they may also be cells with relatively greater proliferative potential than the PLC and are thus likely to be stem cells. PMID- 10465013 TI - Liver cells can spontaneously resume proliferation in long-term quiescent primary cultures. AB - We report here data on the spontaneous resumption of proliferation in long-term primary cultures and we show that the proliferating areas are neoplastic. Normal rat hepatocytes were explanted in serum-supplemented Ham F12 medium and maintained over 8 months without transfer. The cells remained quiescent for the first 10 weeks and they were not tumorigenic when injected into nude mice. Later, without any modification of the culture conditions or transfer, progressive changes spontaneously occurred. Foci of dividing cells were detected, some displaying gamma-glutamyl-transpeptidase (gamma-GT) activity and F-actin fragmentation. These proliferating foci overcame the quiescent population. When injected into nude mice, the 15-week-old primary cultures were highly tumorigenic, with a 3-6 week latency for tumour formation. Furthermore, a cell line was derived from a primary culture started with a liver carcinogen promoter (biliverdin-enriched medium). This cell line proliferated rapidly and differed from a liver epithelial line, also established from our primary cultures, in its 1 karyotype (hyperploidy and translocation on chromosome 3), 2 requirement for arginine to proliferate, 3 gamma-GT positive reaction correlated to changes in actin fibre pattern, 4 sensitivity to protease inhibitors (i.e. alpha2 macroglobulin, PMSF) and 5 tumorigenicity. Long-term primary cultures and the karyotypically defined cell line are useful tools for further studies on in vitro genetic deviations. PMID- 10465014 TI - Changes in oncogene expression in ascite tumour cells during ageing. AB - The expression of two oncogenes, c-myc and c-fos, was studied in an ascitic tumour (ATPC+) at different times after implantation. The specific mRNA synthesis was analysed by Northern blot analysis. The presence of the oncogene proteins was shown by immunofluorescence using flow cytometry and referred to the distribution of the cells in the different cell phases. The results show that both oncogenes are expressed by ATPC+ tumour cells. c-myc is expressed 5, 8 and 12 days after implantation, although with a different intensity, and the protein is mainly present in S or S+G2 phase cells. The c-fos oncogene is expressed only 12 days after tumour implantation and the cells labelled with the specific antibody are mainly in G1 phase. We conclude that c-myc is principally correlated with proliferative activity, whereas c-fos is expressed by non-cycling cells. PMID- 10465015 TI - Gompertzian re-evaluation of the growth patterns of transplantable mammary tumours in sialoadenectomized mice. AB - The method of the recursion formula of the Gompertz function (Bassukas & Maurer Schultze 1988) has been applied to analyse tumour growth data taken from the literature; namely the growth perturbation of transplantable mammary tumours in sialoadenectomized mice with or without subsequent epidermal growth factor substitution (results on two mouse strains, C3H or SHN, have been reported; Inui, Tsubura & Morii 1989). The recursion formula of the Gompertz function fits growth curves to all seven sets of data well (P > 0.05 for lack of fit test). The growth pattern of the tumours in the unperturbed hosts is Gompertzian and does not change if tumours are transplanted in sialoadenectomized mice, although the starting specific growth rate decreases in C3H mice. However, if sialoadenectomy is carried out after tumour inoculation, a complex alteration of the tumour growth evolves: tumour growth does not simply decelerate but it also shifts from the conventional Gompertzian to an exponential or even 'hyperexponential' growth pattern, i.e. with an accelerating specific growth rate. Some theoretical mechanisms of this alteration, as well as the differences between the present Gompertzian analysis and a previously published Verhulstian analysis of part of the same data (Leith, Harrigan & Michelson 1991), are discussed. It is concluded that the quantitative analysis of tumour growth patterns by the method of the difference equation of the Gompertz function presently applied may substantially contribute to the improvement of the interpretation of perturbations of tumour growth--irrespective of their genesis. In contrast to the application of some a priori fixed growth function, e.g. the Verhulstian one, the present method can quantitatively interpret different growth patterns and their classification on the basis of linear regression analysis. PMID- 10465016 TI - Evaluation of the growth rate of MCF-7 breast cancer multicellular spheroids using three mathematical models. AB - Growth data on 60 multicellular spheroids of MCF-7 human breast cancer cells were fitted, on an individual basis, by the Gompertz, Bertalanffy and logistic equations. MCF-7 spheroids, initiated and grown in medium containing oestrogens, exhibited a growth rate that decreased continuously as spheroid size increased. Plots of spheroid volume v. time generated sigmoid curves that showed an early portion with an approximately exponential volume increase; a middle region or retardation phase characterized by a continuously decreasing growth rate; and, finally, a late segment or plateau phase approaching zero growth rate, that permitted an estimate of the maximum spheroid size (Vmax). Growth curves generated by MCF-7 spheroids under different experimental conditions (hormones, drugs and radiation exposures) can be compared after normalization. Linearized forms of the fitted Gompertz curves provided a convenient way to express differences in growth rate. PMID- 10465017 TI - The effects of vinblastine in assessment of the influence of age on proliferative activity of murine palate and footpad epithelium. AB - Data concerning changes in the rate of cell proliferation of stratified epithelia with increasing age are conflicting. In the present study young (3-month-old) and old (22-month-old) C57Bl/6NNia male mice were injected intraperitoneally with 2, 3, 4 or 8 mg vinblastine sulfate/kg body weight and killed after 1.5, 3, 4.5 or 6 h. The number of arrested metaphase figures per 1000 basal cells was counted in histological sections. Data were analysed using a multivariate analysis of variance. There was a significant difference between the accumulation of mitotic figures in footpad epidermis and palate epithelium and both tissues contained an increased number of mitotic figures with increasing periods of accumulation at all dose levels. In the footpad epidermis neither the age of the animal nor the dose of vinblastine had a significant effect on the number of mitotic figures. In contrast, for palate epithelium the accumulation of mitotic figures was significantly less in the old mice compared with the young mice and at a dose of vinblastine of 2 mg/kg compared with the higher doses. There was a statistically significant interaction between the dose of vinblastine and its period of action. It was concluded that the different tissues manifest a differential sensitivity to vinblastine and that only palate epithelium showed a significant reduction in proliferative activity with age. PMID- 10465018 TI - Proliferation and immunoglobulin secretion of lymphoblastoid cell lines are differently affected by soluble cytokines. AB - The aim of our study was to investigate whether supernatant from lipopolysaccharide-activated monocytes (monocyte-factor) and/or cytokines could enhance secretion of human monoclonal antibodies specific to HLA antigens produced by Epstein-Barr virus lymphoblastoid cell lines (EBV-LCLs). In a low cell density culture system, the monocyte-factor significantly stimulated cell growth of three monoclonal and two polyclonal EBV-LCLs while no enhancement of immunoglobulin production was observed. The enhancement of proliferation was completely neutralized by an antiserum to human IL-6 suggesting that IL-6 was required for the stimulation of growth of LCLs. The effect of cytokines on proliferation showed large variations among the cell lines, with IL-1beta generally inducing the highest response. Of the cytokines tested, only IL-2 was able to enhance total immunoglobulin secretion due to the induction of a higher production of light chains. The specific anti-HLA activity was slightly increased by IL-10 although this cytokine had no effect on total immunoglobulin concentration or proliferation. PMID- 10465019 TI - The cell cycle effects and induction of apoptosis by 5-bromouridine in cultures of human leukaemic MOLT-4 and HL-60 cell lines and mitogen-stimulated normal lymphocytes. AB - 5-Bromouridine (BrUrd) and several analogues of this antimetabolite exhibit antiviral activity and are potent inhibitors of the human immunodeficiency virus. The antitumour activity of BrUrd, however, in comparison with 5-fluorouridine or 5-fluorouracil, is less pronounced. Because BrUrd is incorporated into RNA and can be detected immunocytochemically and analysed by cytometry, it may prove useful as an RNA precursor to assay cell transcriptional activity. The aim of the present study was to evaluate the cell cycle effects of BrUrd incorporation in human lymphocytic leukaemic MOLT-4 and HL-60 cells and mitogenically stimulated normal peripheral blood lymphocytes. BrUrd suppressed cell proliferation and induced cell death; when measured after 72 h of treatment, the LD50 was 10 and 20 microM for HL-60 and MOLT-4, respectively, and LD90 was 100 microM, for both cell lines. BrUrd was maximally incorporated by the cells progressing through S phase of the cycle and the S phase cells were most severely perturbed by the drug, which was detected in RNA but not in DNA. Suppression of the S phase traverse of MOLT-4, HL-60 and normal lymphocytes was seen at > or = 30 microM BrUrd concentration. Also sensitive was the cell traverse through G2+M which, in the case of lymphocytes, HL-60 and MOLT-4 cells, was perturbed at > or = 30, > or = 200 and 500 microM BrUrd concentrations, respectively. Transition of lymphocytes from G0 to G1 was little affected at < 100 microM, and, although suppressed, was still observed even at 500 microM BrUrd concentration. Apoptosis of HL-60 cells and lymphocytes was observed at > or = 50 microM BrUrd after 24 h of incubation; death of MOLT-4 cells had mixed features of apoptosis and necrosis and resembled the 'mitotic' or 'reproductive cell death' as described in other cell systems. The cytostatic and cytotoxic effects of BrUrd should be taken into consideration when using this antimetabolite as an antiviral agent in the clinic or as an RNA precursor in assays of cell transcriptional activity. PMID- 10465020 TI - Cell kinetic studies in mouse tongue mucosa by autoradiographic, immunohistochemical and flow cytometric techniques. AB - A number of techniques, including autoradiography after in vivo administration of tritiated thymidine ([3H]dT), immunohistochemistry after in vivo administration of bromodeoxyuridine (BrdUrd), and flow cytometry (FCM) with and without BrdUrd detection were compared in the epithelium of ventral mouse tongue. Investigation of the diurnal proliferative rhythm by immunohistochemical detection of incorporated BrdUrd with different primary antibodies in combination with the alkaline-phosphatase anti-alkaline-phosphatase technique, the peroxidase-anti peroxidase method, and an indirect method with a polyclonal peroxidase-conjugated secondary antibody yielded results similar to standard autoradiography. Preparation of single cell suspensions for flow cytometry was not successful. A maximum yield of about 8.5% of the original cell number was achieved by ultrasound disintegration in combination with trypsin and dithioerythrol treatment, but neither a G0/G1 peak nor a G2 + M peak was observed in DNA histograms. A better yield of about 38% of the original nuclei number was obtained by preparation of suspensions of nuclei using citric acid and the detergent Tween 20 in combination with magnetic stirring. Both S-phase index and BrdUrd labelling index could be determined by FCM and showed the normal diurnal variations. However, the BrdUrd labelling index in suspensions of nuclei was significantly higher than the labelling index determined after immunohistochemistry. The FCM S-phase index at times of day with low DNA synthesizing activity was higher than the BrdUrd index, indicating a fraction of unlabelled S-phase cells. In conclusion, detection of incorporated BrdUrd in oral mucosa by immunohistochemical techniques or flow cytometry is feasible and provides a useful tool for fast measurements of proliferation. PMID- 10465021 TI - Seasonal variation in proliferative response and subpopulations of lymphocytes from mice housed in a constant environment. AB - A seasonal variation in the proliferative response to mitogens and in the proportion of splenic lymphocyte subpopulations was found in mice housed in a constant environment. The lymphoproliferative responses to T-cell and B-cell mitogens reached maximum values in autumn and summer. Identification of lymphocyte subpopulations by flow cytometry demonstrated that the proportion of T cytotoxic-suppressor (Tcs) lymphocytes was significantly higher in autumn and summer than in spring and winter. However, the proportion of B lymphocytes was significantly lower in spring than in the other three seasons, whereas the proportions of T and T helper (Th) cells did not show any seasonal variation. On the other hand, we observed a significant correlation between the level of mitogenic responsiveness and the proportion of Tcs cells, but not between the former and the proportions of B, T or Th cells. These data suggest that the seasonal variation in murine lymphoproliferative responses may depend on the cyclic changes in the proportion of Tcs lymphocytes; these changes, in turn, may be predetermined by the inherent internal biological rhythms of the animal. PMID- 10465022 TI - Repair, redistribution and repopulation in V79 spheroids during multifraction irradiation. AB - Development of predictive assays for measuring tumour radiosensitivity has generated much recent interest, particularly with the recognition that tumour cell survival at doses of about 2 Gy may correlate well with tumour curability. Clinical data, however, suggest that overall treatment time may be of considerable significance in radioresponsive tumours, especially for rapidly growing tumours capable of accelerated repopulation. Because neither factor can be repeatedly assessed in human tumours, we used cells growing as multicell spheroids to determine whether the initial radiation response would be predictive for multifraction exposures, or whether other factors including repopulation rate should be considered. Potential problems of hypoxia and reoxygenation were avoided by using small spheroids which had not yet developed radiobiologically hypoxic regions. Repair and redistribution dominated the responses in the first two or three exposures, with repopulation playing a minor role. As the fractionation schedule was extended, however, repopulation between fractions largely determined the number of viable cells per spheroid. We conclude that the radiation response of cells from untreated spheroids provides a general indication of net sensitivity, but that repair and redistribution produces considerable variation in radiosensitivity throughout a fractionation protocol. Ultimately, repopulation effects may dominate the multifraction response. PMID- 10465023 TI - Cell genealogies in a plant meristem deduced with the aid of a 'bootstrap' L system. AB - The primary root meristem of maize (Zea mays L.) contains longitudinal files of cells arranged in groups of familial descent (sisters, cousins, etc.). These groups, or packets, show ordered sequences of cell division which are transverse with respect to the apico-basal axis of the root. The sequences have been analysed in three zones of the meristem during the course of the first four cell generations following germination. In this period, the number of cells in the packets increases from one to 16. Theoretically, there are 48 possible division pathways that lead to the eight-cell stage, and nearly 2 x 10(6) that lead to the 16-cell stage. However, analysis shows that only a few of all the possible pathways are used in any particular zone of the root. This restriction of pathways results from inherited sequences of asymmetric cell divisions which lead to sister cells of unequal length. All possible division pathways can be generated by deterministic 'bootstrap' L-systems which assign different lifespans to sister cells of successive generations and hence specify their subsequent sequence of divisions. These systems simulate propagating patterns of cell divisions which agree with those actually found within the growing packets that comprise the root meristem. The patterns of division are specific to cells originating in various regions of the meristem of the germinating root. The importance of such systems is that they simulate patterns of cellular proliferation where there is ancestral dependency. They can therefore be applied in other growing and proliferating systems where this is suspected. PMID- 10465024 TI - A possible interpretation of some colony forming assays: 'developmental tree' model. AB - The characteristic form of the probability distributions of in vitro grown colonies is analysed and experimental results are shown to be well approximated by an exponential dependence. It is argued that this type of dependence is very frequent in nature and does not provide sufficient evidence to resolve the dilemma whether the development of haemopoietic lineages proceeds according to a schema of 'progressive and stochastic restriction' or more limiting 'developmental tree' with some random control process involved. Detailed analysis of the experimental conditions and results is needed to provide insight into collective phenomena in cellular proliferation and differentiation. At present, the complexity of the phenomena involved prevents unique interpretation of the experimental distributions. Nevertheless, one particular quantitative model, based on a developmental tree concept, yields a set of the theoretical probability distributions, which are in a very good agreement with the experimental results. PMID- 10465025 TI - A dual block to cell cycle progression in HL60 cells exposed to analogues of vitamin D3. AB - The physiologically active form of vitamin D3, 1,25-dihydroxy-vitamin D3 (1,25(OH)2D3), induces differentiation of several types of myeloid leukaemia cells. The acquisition of monocyte-like phenotype is accompanied by slower progression through the cell cycle, and G1 block has been reported to be the basis of this effect. It is shown here that human promyelocytic leukaemia HL60 cells treated with analogues of vitamin D3 which are potent inducers of monocytic differentiation have an additional cell cycle block. Exposure to 10(-7) M 1,25(OH)2D3 or 1,25-(OH)2-16-ene-D3 resulted in monocytic differentiation and the expected G1 block evident at approximately 48 h in a rapidly differentiating variant of HL60 cells (HL60-G), and at 96 h in the more slowly differentiating HL60-240 cells. In addition, a G2+M block was noted at approximately 72 h in HL60 G and HL60-240 cells. Exposure to vitamin D3 analogues also markedly increased the number of dikaryons, suggesting that cytokinesis was impaired more than karyokinesis. Treatment with a third analogue 25-hydroxy-16,23-diene-D3 produced little differentiation and had minimal effects on the cell cycle parameters. These findings indicate that vitamin D3 analogues regulate cell proliferation by control of the transition of G1 and G2+M phases, reminiscent of the cdc2/CDK2 type of cell cycle control. PMID- 10465026 TI - Leukaemic peripheral blood plasma and bone marrow plasma: comparison of influence on lymphocyte proliferation. AB - Peripheral blood plasma from some children with untreated acute lymphoblastic leukaemia (ALL) exerted an inhibitory effect in vitro on phytohaemagglutinin induced lymphocyte transformation of normal peripheral blood lymphocytes. This occurred at concentrations beyond that required for optimal response as judged by reduction of blast cell formation and tritiated thymidine and tritiated uridine incorporation into DNA and RNA, respectively. In contrast, bone marrow plasma from these patients was non-inhibitory or contained significantly less inhibitory activity. Bone marrow plasma from the majority of healthy controls was superior to their peripheral blood plasma in enhancing phytohaemagglutinin-induced mitogenesis. The difference between an individual's bone marrow- and peripheral blood-derived plasma in enhancing proliferation of patient and healthy control cells was significantly greater amongst the patients than the healthy control group; this was attributed mainly to the increased inhibitory activity of ALL peripheral blood plasma compared with normal plasma. Medium conditioned by phytohaemagglutinin-stimulated normal peripheral blood lymphocytes was effective in neutralizing the inhibitory activity of ALL peripheral blood plasma. Taken together, these in vitro results are at least suggestive that in vivo, in healthy subjects, the rapidly proliferating cells in the bone marrow and the 'resting' blood cells in the circulation may be under the influence of a fine balance of different types and/or levels of humoral growth stimulatory and inhibitory factors and that in ALL an unstable balance of these factors exists. The decreased proliferation of circulating blast cells compared with bone marrow blasts in ALL may be attributed, at least in part, to exposure to the different levels of inhibitor(s) in the circulation and bone marrow as demonstrated in vitro by our results. PMID- 10465027 TI - The role of the IGF-I receptor in the growth and transformation of mammalian cells. AB - Recent developments in the molecular biology of the insulin-like growth factor I (IGF-I) receptor have clarified its role in cellular growth and transformation. Although cells homozygous for a targeted disruption of the IGF-I receptor genes can grow in serum-supplemented medium, the IGF-I receptor is required for optimal growth, and is required equally in all phases of the cell cycle. The receptor plays an even more stringent role in cellular transformation and tumorigenicity, which seem to be dependent on its normal expression in several cell types. The expression of both the IGF-I receptor and its ligands is regulated by other growth factors (especially PDGF and EGF), by oncogenes (like SV40 T antigen and c myb) and by tumour suppressor genes (like WT1 and RB). The picture emerging from these studies is that several transforming agents may exert their growth promoting effects through the direct or indirect activation of the IGF autocrine loop. PMID- 10465028 TI - Analysis of growth of multicellular tumour spheroids by mathematical models. AB - We wished to determine the applicability of previously proposed deterministic mathematical models to description of growth of multicellular tumour spheroids. The models were placed into three general classes: empirical, functional and structural. From these classes, 17 models were applied systematically to growth curves of multicellular tumour spheroids used as paradigms of prevascular and microregional tumour growth. The spheroid growth curves were determined with uniquely high density of measurements and high precision. The theoretical growth curves obtained from the models were fitted by the weighted least-squares method to the 15 measured growth curves, each corresponding to a different cell line. The classical growth models such as von Bertalanffy, logistic and Gompertz were considered as nested within more general models. Our results demonstrate that most models fitted the data fairly well and that criteria other than statistical had to be used for final selection. The Gompertz, the autostimulation and the simple spheroid models were the most appropriate for spheroid growth in the empirical, functional and structural classes of models, respectively. We also showed that some models (e.g. logistic, von Bertalanffy) were clearly inadequate. Thus, contrary to the widely held belief, the sigmoid character of a three or more parameter growth function is not sufficient for adequate fits. PMID- 10465029 TI - The fraction of cells in G1 with bound retinoblastoma protein increases with the duration of the cell cycle. AB - The retinoblastoma gene product (pRB) is a nuclear phosphoprotein with growth suppressing effects. During early G1 phase, pRB is underphosphorylated and bound in the nucleus. The association between the duration of the cell cycle/G1 phase and the fraction of cells in G1 with bound pRB was studied in the human pre-B cell line Reh. The cell-cycle duration was varied by growing cells at different concentrations (25, 10, 2, 0.5 and 0%) of fetal calf serum (FCS); pRB binding was studied by flow cytometry. The culture doubling time increased from 21 h in 25% FCS to 54 h in 0.5% FCS. Cell death occurred in the absence of FCS, and the culture doubling time therefore could not be defined. The fraction of cells in G1 did not change significantly with decreasing FCS concentration (0.47 in 25% FCS, 0.52 in 0% FCS). In contrast, the fraction of G1 cells with bound pRB increased from 0.12 in 25% FCS to 0.65 in 0% FCS. Continuous labelling with bromodeoxyuridine demonstrated that the growth fraction was close to unity at all FCS concentrations down to 0.5%, hence, the duration of the cell cycle was equal to the culture doubling time under these conditions. The duration of early G1 phase (where pRB is underphosphorylated and bound) increased 10-fold, while the duration of late G1 phase increased twofold, for Reh cells grown in 0.5% FCS compared with cells grown in 25% FCS. The increase in the duration of late G1, and the increased S and G2+M phase transit times, indicate that other factors, in addition to pRB kinase activity, regulate the duration of G1 and the cell cycle of serum-deprived Reh cells. PMID- 10465030 TI - A model for regulation of the cell cycle incorporating cyclin A, cyclin B and their complexes. AB - A mathematical model for the cell cycle is proposed that incorporates the known biochemical reactions involving both cyclin A and cyclin B, the interactions of these cyclins with cdc2 and cdk2, and the controlling effects of cdc25 and weel. The model also postulates the existence of an as yet unknown phosphatase involved in the formation of maturation promoting factor. The model produces solutions that agree qualitatively with a wide variety of experimentally observed cell cycle behaviour. Conditions under which the model could explain the initial rapid divisions of embryonic cells and the transition to the slower somatic cell cycle are also discussed. PMID- 10465031 TI - Origin and evolution of binucleate cells in cultures of HEp-2 cells. AB - The origin and evolution of binucleate cells in cultures of HEp-2 cells have been studied by means of interval photography and time-lapse video-recording. Binucleate cells most frequently formed by the fusion of two sister cells born in a previous mitosis. The study of binucleate cells has shown that they are a cellular type able to successfully undergo mitosis. However, the mitosis may be bipolar, tripolar or multipolar. The daughter cells arising from these divisions do not follow a clear pattern in the number of nuclei they have, instead showing a wide range of possibilities. PMID- 10465032 TI - Comparison of the beta-glucuronidase assay and the conventional method for identification of Escherichia coli on eosin-methylene blue agar. AB - The purpose of this study was to compare the IMViC (indole, methyl red, Voges Proskauer and citrate utilization) tests with the beta-glucuronidase (GUD) assay for the identification of suspect Escherichia coli on Levine's eosin-methylene blue (EMB) agar. After testing 258 suspect E. coli colonies from raw meat and meat products, 163 and 44 were found to be E. coli and non-E. coli, respectively, by both methods. Nine isolates were IMViC positive (i.e., + + - - or - + - -) but GUD negative; among these isolates, six were confirmed to be E. coli by API 20E (bioMerieux, Marcy-I'Etoile, France) with the remaining three being non-E. coli. There were 42 isolates that were IMViC negative but GUD positive; among these isolates, seven were pure E. coli cultures, 33 were mixed cultures containing E. coli, and the remaining two were Proteus spp. The sensitivities for the identification of E. coli on EMB were 80.9% (169/209) and 97.1% (203/209), respectively, by the IMViC tests and GUD assay; whereas the specificities were 93.9% (46/49) and 95.9% (47/49), respectively, by the IMViC tests and GUD assay. It is proposed that the GUD assay can be an effective alternative to the conventional IMViC tests for the identification of suspect E. coli on EMB. PMID- 10465033 TI - Provision of lactose to molting hens enhances resistance to Salmonella enteritidis colonization. AB - Older leghorn hens, more than 50 weeks of age, were divided into three groups designated 1, unmolted controls; 2, molted; or 3, molted treated with lactose. Forced molt was induced by 14 days of feed removal. Lactose was provided to the hens in group 3 as 2.5% (wt/vol) of the daily drinking water. Each hen in all groups was challenged orally with 10(5) Salmonella enteritidis (SE) cells on day 7 of feed removal. The study was repeated in three replicated trials. The concentrations of acetic, propionic, and total volatile fatty acids (VFA) in the cecal contents of the molted hens in groups 2 and 3 decreased significantly (P < 0.05) on days 6 and 14 of molt compared with the unmolted controls. Forced molt had no apparent effect on pH or on the oxidation-reduction potential of the ceca. Compared to the unmolted controls, SE cecal and spleen and liver colonization was significantly increased (P < 0.05) in the molted hens in group 2. Compared to the molted hens in group 2, SE cecal and spleen and liver colonization was significantly decreased (P < 0.05) in two of three trials in the hens in group 3 provided with lactose. The results suggested that the increased susceptibility of molting hens to SE colonization may be associated with decreased fermentation and production of VFA by cecal bacteria or by a depletion of the number of VFA producing bacteria present in the ceca. The results further suggest that providing lactose in the drinking water during molting may significantly enhance resistance to SE colonization. PMID- 10465034 TI - Attachment of Salmonella choleraesuis choleraesuis to beef muscle and adipose tissues. AB - The attachment of Salmonella choleraesuis subsp. choleraesuis ATCC 15790 to beef muscle and adipose tissues was investigated. S. choleraesuis was found to adhere in higher numbers to muscle than to fat. The charge and the hydrophobicity of the surface of S. choleraesuis were evaluated by measurement of electrophoretic mobility, the contact angle with water, adhesion to hexadecane, and hydrophobic interaction chromatography. The overall negative charge of S. choleraesuis was masked by the high electrolyte concentration in the attachment medium (0.15 M phosphate-buffered saline). This bacterium was shown to possess a hydrophilic surface. Electrostatic interactions do not affect the attachment of S. choleraesuis to both lean and fat tissue, and there was no evidence for a role of hydrophobic interactions. However, the attachment of S. choleraesuis was reduced by 90% after mechanical removal of the flagella or after treatment of the bacteria with specific antiflagella serum. This reduction was attributed to a loss of bacterial mobility leading to a reduction in the number of cells reaching the tissue during the period of contact. Treatment of the tissue with a concentrated suspension of flagella or treatment of the bacteria with antisomatic serum (OMD) did not reduce the attachment of S. choleraesuis to tissues, indicating an absence of specific attachment sites for flagella or antigen O on the beef tissue surface. PMID- 10465035 TI - Monoclonal antibodies and an indirect ELISA for detection of psychrotrophic bacteria in refrigerated milk. AB - Monoclonal antibodies generated against live cells of Pseudomonas fluorescens have been used in an indirect ELISA format for the detection of Pseudomonas spp. and related psychrotrophic bacteria in refrigerated milk. The immunorecognition of monoclonal antibodies adsorbed to bacteria bound to the wells of a microtiter plate was performed with rabbit anti-mouse immunoglobulins conjugated to horseradish peroxidase. Subsequent enzymic conversion of the substrate resulted in distinct absorbance differences when assaying milk samples containing psychrotrophic bacteria in the range 10(5) to 10(9) CFU ml(-1) . The detection threshold for the ELISA assay developed in this work is 10(5) CFU ml(-1). PMID- 10465036 TI - Bactericidal effect of enterocin 4 on Listeria monocytogenes in a model dairy system. AB - Enterococcus faecalis INIA 4 produced the bacteriocin enterocin 4 during growth in raw ewe's milk at 30 degrees C. Enterocin activity reached 2,200 to 3,600 AU/ml after 8 h, with a 1 to 8% (vol/vol) level of inoculum from an 18-h culture. An enterocin activity of 500 AU/ml significantly decreased counts of Listeria monocytogenes Ohio when incubated for 6 h in a model system consisting of filtrates from cultures of E. faecalis INIA 4 in raw ewe's milk, at pH 6.0 and 30 degrees C. However, an enterocin activity of 2,400 AU/ml was needed in the same conditions to significantly decrease counts of L. monocytogenes Scott A. All 22 wild L. monocytogenes strains isolated from ewe's milk and tested were inhibited by a filtrate containing 400 AU/ml of enterocin 4. Incubation in the filtrate for 6 h significantly lowered counts of 16 L. monocytogenes strains, and incubation for 24 h, counts of 21 strains. PMID- 10465037 TI - Effect of high hydrostatic pressure on Listeria innocua 910 CECT inoculated into Ewe's milk. AB - Ewe's milk standardized to 6% fat was inoculated with Listeria innocua 910 CECT at a concentration of 10(7)CFU/ml and treated by high hydrostatic pressure. Treatments consisted of combinations of pressure (200, 300, 350, 400, 450, and 500 MPa), temperature (2, 10, 25, and 50 degrees C), and time (5, 10, and 15 min). To determine numbers of L. innocua, listeria selective agar base with listeria selective supplement and plate count agar was used. Low-temperature (2 degrees C) pressurizations produced higher L. innocua inactivation than treatments at room temperatures (25 degrees C). Pressures between 450 and 500 MPa for 10 to 15 min were needed to achieve reductions of 7 to 8 log units. The kinetics of destruction of L. innocua were first order with D-values of 3.12 min at 2 degrees C and 400 MPa and 4 min at 25 degrees C and 400 MPa. A baroprotective effect of ewe's milk (6% fat) on L. innocua was observed in comparison with other studies using different media and similar pressurization conditions. PMID- 10465038 TI - Airborne bacteria and carcass contamination in slaughterhouses. AB - Microbiological contamination of air and carcasses was studied in four slaughterhouses by using impactor samples taken at the back-splitting and weighing areas and by sampling carcasses with the swabbing method. Air flow was determined by an air-flow detector, and the movement of workers was observed. The air contamination level in the back-splitting areas (2.25 log CFU/100 liters of air) was generally higher than that in the weighing areas (2.03 log CFU/100 liters of air). Associations between the microbiological contamination of air and carcasses with the movements of workers were found. Layout of the slaughtering line was shown to be important in decreasing airborne contamination. Separation of the clean and unclean parts of the line as well as separation of the weighing area from the other clean parts of the line decreased the contamination level. It appears that airborne bacteria have an important role in carcass contamination. PMID- 10465040 TI - Characteristics of low-and high-fat beef patties: effect of high hydrostatic pressure. AB - The purpose of this study was to analyze the consequences of applying high pressures (100 and 300 MPa for 5 or 20 min) on characteristics such as water- and fat-binding properties, texture, color, microstructure, and microbiology of low fat (9.2%) and high-fat (20.3%) beef patties. In nonpressurized patties, the low fat product exhibited significantly poorer (P < 0.05) binding properties and higher (P < 0.05) Kramer shear force and Kramer energy than did high-fat patties. Although high pressure did not clearly influence the binding properties of low- and high-fat beef patties, it did produce a rise in the Kramer shear force and energy which were more pronounced at 300 MPa. High pressures altered patty color, the extent of alteration depending on fat content, pressure, and pressurizing time. Pressurizing high- and low-fat beef patties at 300 MPa not only produced a lethal effect (P < 0.05) on microorganisms, but caused sublethal damage as well. PMID- 10465039 TI - Effectiveness of sanitation with quaternary ammonium compound or chlorine on stainless steel and other domestic food-preparation surfaces. AB - The relative ability of various materials used for domestic and/or food-service sinks and countertops to be sanitized was determined. Both smooth (unused) and abraded surfaces were tested by exposure to 200 mg of quaternary ammonium compound per liter or 200 mg of sodium hypochlorite per liter. Surface materials tested included mechanically polished (type 304, #4 finish) and electropolished stainless steel, polycarbonate, and mineral resin. Surfaces were prepared for testing by allowing attachment of a Staphylococcus aureus culture for 4 h to achieve an initial attached population of 10(4) to 10(5) CFU/cm2. The test procedure involved immersion of the surface in sanitizer solution followed by wiping with a sanitizer-saturated cloth. Residual staphylococci were detected by overlaying agar directly on the treated surface. Results indicated that the stainless steels and the smooth polycarbonate, which had 0.5 log CFU/cm2 or fewer of residual staphylococci, were more readily sanitized by quaternary ammonium compound than were either the mineral resin surfaces, which had nearly 2.0 log CFU/cm2 of residual staphylococci, or the abraded polycarbonate which had nearly 1.0 log CFU/cm2 of residual staphylococci. Chlorine was most effective on the mechanically polished stainless steel, the unabraded electropolished stainless steel, and the polycarbonate surfaces, reducing cell populations to less than 1.0 log CFU/cm2. Chlorine was less effective on abraded electropolished stainless steel and mineral resin surfaces, where populations remained greater than 1.0 log CFU/cm2. Sanitation with quaternary ammonium compound or chlorine reduced S. aureus populations more than 1,000-fold on all surfaces except unabraded mineral resin. PMID- 10465041 TI - Influence of processing schemes on indicative bacteria and quality of fresh aquacultured catfish fillets. AB - Fresh aquacultured catfish fillets were obtained from three processors using different processing protocols in summer, autumn, winter, and spring and evaluated for microbial quality. Twenty freshly processed fillets were randomly selected and each fillet was placed in a sterile polyethylene bag. The fillets were transported on ice-pack overnight by air immediately after processing. Five fillets were randomly selected for microbial assays. Each fillet was weighed and an equal volume of sterile 0.1% peptone water at 0 to 1 degrees C was added aseptically. The fillet was massaged (or rinsed) for 120 s and the rinse was used to determine microbial quality. Aerobes (incubation at 35 degrees C for 48 h) and psychrotrophs (incubation at 20 degrees C for 96 h) were enumerated using 3M Petrifilm Aerobic Count plates. Escherichia coli (incubation at 35 degrees C for 24 to 48 h) and total coliforms (incubation at 35 degrees C for 24 to 48 h) were enumerated on 3M Petrifilm E. coli Count plates. Staphylococcus aureus counts were determined on Baird-Parker agar (incubation at 35 degrees C for 48 h). Significant differences (P < or = 0.05) in aerobic, psychrotrophic, total coliform, E. coli, and S. aureus counts due to temperature effects during production and variations in processing protocols were observed. E. coli and S. aureus counts were significantly different during the four seasons. E. coli and S. aureus counts were high during summer and low during winter weather. There was a significant difference (P < or = 0.05) in aerobic, psychrotrophic, and total coliform counts among the three processors during warm weather; however, these differences were significantly (P < or = 0.05) reduced in cold weather. PMID- 10465042 TI - Natural milk cultures for the production of Argentinian cheeses. AB - Samples (32) of natural milk cultures used in the Santa Fe, Argentina, area for soft and semihard cheese production were examined. The microbial composition (including lactic acid microflora characterization) and technological parameters (acidifying and proteolytic activities) were evaluated. The cultures contained mainly thermophilic lactic acid bacteria, identified as Streptococcus salivarius subsp. thermophilus (96.8% of the total strains) and Enterococcus spp. The strains showed a low proteolytic activity. The isolates of S. salivarius subsp. thermophilus exhibited a widespread phage resistance. The nonlactic microflora comprised coliforms, yeasts, spore-forming bacteria and lactate fermentative bacteria. The samples showed an acidity level from 0.38 to 0.69% lactic acid (pH from 4.25 to 5.75). The acidifying activity was optimal at 45 degrees C. The advantages and disadvantages of the employment of natural milk starters are discussed. PMID- 10465043 TI - A competitive enzyme-linked immunosorbent assay for detection of bovine milk in ovine and caprine milk and cheese using a monoclonal antibody against bovine beta casein. AB - A competitive ELISA (enzyme-linked immunosorbent assay) was performed to detect and quantify bovine milk in ovine and caprine milk and cheese using a monoclonal antibody (AH4 MAb) against bovine beta-casein. Ovine or caprine milk and cheese containing bovine milk were added simultaneously with the AH4 MAb to the wells of a microtiter plate that had been previously sensitized with commercial bovine beta-casein. The bovine caseins in milk or cheese samples compete with the bovine beta-casein bound to the plate for the AH4 MAb binding sites. Further immunorecognition of AH4 MAb bound to the bovine beta-casein immobilized onto the plate was attained with rabbit anti-mouse immunoglobulin conjugated to peroxidase. Subsequent enzymic conversion of the substrate showed clear differences in absorbance values during assay of mixtures of ovine and caprine milk and cheese containing various amounts of bovine milk. The competitive ELISA developed in this work allows the quantitative detection of bovine milk in ovine and caprine milk and cheese samples in the range of 0.5 to 25% of substitution. PMID- 10465044 TI - Antimicrobial activity of sulfur compounds derived from cabbage. AB - Selected sulfur compounds found in cabbage and its fermentation product, sauerkraut, were tested for minimum inhibitory concentration (MIC) against growth of 15 species of bacteria and 4 species of yeasts. S-Methyl-L-cysteine sulfoxide, sinigrin, and dimethyl sulfide at 500 ppm were not inhibitory to any of the bacteria and yeasts tested. Dimethyl disulfide at 500 ppm retarded some, but did not prevent growth of any of the test microorganisms. Dimethyl trisulfide had an MIC to bacteria of 200 ppm and to yeast of 20 ppm. Methyl methanethiosulfinate had an MIC between 50 and 200 ppm for all bacteria, and between 6 and 10 ppm for all yeasts tested. Methyl methanethiosulfonate had an MIC between 20 and 100 ppm for bacteria and between 50 and 500 ppm for yeasts. Allyl isothiocyanate had an MIC between 50 and 500 ppm for bacteria and between 1 and 4 ppm for yeasts. Methyl methanethiosulfinate was 10 to 100 times more inhibitory against Listeria monocytogenes at pH values of 5, 6, and 7 and was much less influenced by pH than was sodium benzoate. PMID- 10465045 TI - Isolation and identification of antimicrobial furocoumarins from parsley. AB - Photoactive furocoumarins extracted from four varieties of fresh and freeze-dried parsley leaves inhibited a DNA repair-deficient Escherichia coli in a photobiological assay. Using media-modified assays, the human pathogens E. coli O157:H7 and Listeria monocytogenes, the spoilage microorganism Erwinia carotovora, and Listeria innocua were also inhibited. Pseudomonas fragi was not inhibited. Minimum concentrations of Forest Green parsley powder in agar which showed inhibition ranged from 0.12% to 8.0% depending on the microorganism. Ultraviolet light (UV) at 365 nm for 60 min used to photoactivate the furocoumarins in the bioassay had little effect on L. monocytogenes and L. innocua. A slight UV inhibitory effect was detected with E. carotovora. Furocoumarins, psoralen, 8-methoxypsoralen (8-MOP), 5-methoxypsoralen (5-MOP), oxypeucedanin and isopimpinellin were identified using gas chromatography-mass spectrometry. Psoralen, 8-MOP, and 5-MOP were quantified. A difference in relative furocoumarin concentration (weight of furocoumarin per weight of dry parsley leaves) for all varieties of parsley was revealed. The concentration of 5 MOP was significantly greater than 8-MOP (P < 0.05), but not significantly greater than psoralen. Psoralen and 8-MOP were not significantly different in concentration. PMID- 10465046 TI - Acaricide residues in honeys from Galicia (N.W. Spain). AB - Honey samples (101) from Galicia (N.W. Spain) were analyzed by gas chromatography (electron capture and flame ionization) for the presence of acaricides (amitraz, bromopropylate, coumaphos, and fluvalinate). Seventy-three samples were free from detectable residues. Bromopropylate residues were found in 16 samples in levels ranging from 5 to 60 microg/kg. Fluvalinate residues were found in 11 samples in levels ranging from 10 to 40 microg/kg. One sample contained 100 microg of fluvalinate per kg. Neither amitraz nor coumaphos residues were detected. PMID- 10465047 TI - Evaluation of the polymerase chain reaction method for identification of Vibrio vulnificus isolated from marine environments. AB - The polymerase chain reaction (PCR) method for identification of Vibrio vulnificus in the marine environment was evaluated by comparing it to both the conventional and DNA-DNA hybridization methods. Of 13,325 isolates obtained from seawater and sediment samples, and oyster and goby specimens collected from the coastal waters of Tokyo Bay, Japan, only 61 isolates were identified as V. vulnificus on the basis of phenotypic characteristics and the amplification of the cytotoxin-hemolysin gene by the PCR method. All 61 isolates were further confirmed to be V. vulnificus by a DNA-DNA hybridization method and the API 20E system although they were divided into 13 groups on the basis of their API 20E profiles. These results strongly suggest that the PCR method is useful for identification of this organism. PMID- 10465048 TI - Growth of an Aspergillus flavus transformant expressing Escherichia coli beta glucuronidase in maize kernels resistant to aflatoxin production. AB - Kernels of a maize inbred that demonstrated resistance to aflatoxin production in previous studies were inoculated with an Aspergillus flavus strain containing the Escherichia coli beta-D-glucuronidase reporter gene linked to a beta-tubulin gene promoter and assessed for both fungal growth and aflatoxin accumulation. Prior to inoculation, kernels were pin-wounded through the pericarp to the endosperm, pin wounded in the embryo region, or left unwounded. After 7 days incubation with the fungus, beta-glucuronidase activity (fungal growth) in the kernels was quantified using a fluorogenic assay and aflatoxin B content of the same kernels was analyzed. Kernels of a susceptible inbred, similarly treated, served as controls. Results indicate a positive relationship between aflatoxin levels and the amount of fungal growth. However, resistant kernels wounded through the pericarp to the endosperm before inoculation supported an increase in aflatoxin B over levels observed in nonwounded kernels, without an increase in fungal growth. Wounding kernels of the resistant inbred through the embryo resulted in both the greatest fungal growth and the highest levels of aflatoxin B1 for this genotype. Maintenance of resistance to aflatoxin B1 in endosperm-wounded kernels may be due to the action of a mechanism which limits fungal access to the kernel embryo. PMID- 10465049 TI - Antibody isotype responses, infection and re-infection for Schistosoma japonicum in a marshland area of China. AB - Antibody isotype responses to adult worm antigen (AWA) of Schistosoma japonicum and two recombinant proteins (paramyosin (PMY) and a 22 kDa tegumental membrane associated antigen (TEG)) were analyzed in 137 individuals from an area moderately endemic for schistosomiasis in the Dongting Lake region, Hunan Province, China. The prevalence and geometric mean (GM) intensity of infection before the implementation of curative chemotherapy were 28.5% and 234.4 epg, respectively, but 9 months after treatment the prevalence (6.6%) and intensity (38.3 epg) had decreased. There was no significant difference in either the prevalence or intensity of infection between males and females. Specific IgG (total), IgG4, IgG2, IgA and IgE responses to AWA, PMY and TEG were measured by ELISA. Males produced significantly (P < 0.05) more anti-AWA total IgG, IgE, IgA, IgG4 and IgG2 antibodies, and anti-TEG IgG2 antibody than their female counterparts. The OD450 levels of anti-AWA, PMY and TEG antibody isotypes did not present clear age-dependent trends except for peak levels of anti-AWA IgG4 antibodies evident among subjects 20-29 years of age. The total IgG and IgG4 antibody profiles against AWA correlated well with current S. japonicum infections while anti-AWA IgG2, IgA and IgE antibodies did not show such an association. Anti-AWA-specific IgE antibody levels were positively correlated (r = 0.55) with anti-AWA specific IgG4 antibody levels. In addition, the overall percentage of responders (using a cut-off value obtained from normal controls) to all isotypes to AWA were higher than those observed for both the recombinant antigens. Only 18.2%, 16.8% and 7.3% of the study population were IgE responders to AWA, PMY and TEG. A longer follow-up period is required before we can more fully understand the role of IgE, if any, in protective immunity against schistosomiasis japonica. PMID- 10465050 TI - Phospholipid signalling pathways in Trypanosoma cruzi growth control. AB - The role of phospholipids (PLs) in the signal transduction pathways that are activated by a mitogenic stimulus (foetal calf serum) in Trypanosoma cruzi epimastigotes (EPI) was investigated. Only phosphatidylinositol-bis-phosphate was significantly altered in this process. Other phosphoinositides, including major PLs such as phosphatidylcholine and phosphatidylethanolamine, were unaltered. Lysophosphatidic acid, reported to be the primary active substance in effects of serum in other systems, had no mitogenic activity when added to epimastigote cultures. Involvement of phosphoinositide-specific phospholipase C was established using the inhibitors ET-18-OCH3 and U73122, which prevented phosphatidylinositol-bis-phosphate hydrolysis; the latter compound decreased T. cruz proliferation. The intracellular signalling downstream to the phospholipase C was mediated by Ca2+/PL-dependent protein kinase and Ca2+/calmodulin-dependent protein kinase II, judging from the marked decrease in replication caused by the specific inhibitors staurosporine, derythro-sphingosine and KN-93. Previous reports have demonstrated a dual control of cell growth in EPI, whose proliferation is stimulated by the activation of a phospholipase C system and inhibited by activation of an adenylate cyclase system. Investigating this 'cross talk' phenomenon, we observed that an increase in intracellular cAMP inhibited growth mediated by a cAMP-dependent protein kinase, but did not cause PL alterations, and also did not prevent the effect of serum on them. PMID- 10465051 TI - CD4+ and/or gammadelta+ T cells in the liver spontaneously produce IL-4 in vitro during the early phase of Leishmania major infection in susceptible BALB/c mice. AB - Numerous studies on the cytokine production profile in Leishmania major infected susceptible and resistant mice have been carried out to elucidate the mechanisms of healing or non-healing of this infection. However, many methods may have failed to detect the actual cytokine production in the inflammatory foci. To overcome this problems, the ELISPOT assay was used to examine the spontaneous production of IL-4 and IFN-gamma in vitro by mononuclear cells from the spleen, lymph nodes and liver in L. major-infected susceptible BALB/c and resistant C57BL/6 mice. None of these mononuclear cells spontaneously produced IFN-gamma in either mouse strains in vitro in the absence of the corresponding antigen(s). However, liver mononuclear cells from infected BALB/c mice spontaneously produced IL-4 in vitro in as early as 2 weeks after the infection, but this was not observed in C57BL/6 mice. The IL-4 producing liver lymphocytes consisted of CD4+ and/or gammadelta+ T cells and uncharacterized cells. These results suggest that liver lymphocytes play some role in the establishment of Th2 prevalence in susceptible BALB/c mice, based on the importance of IL.4 production in the early phase of L. major infection in establishing Th2 dominance in this parasite susceptible mouse. PMID- 10465052 TI - Characterization of intracellular metabolites of axenic amastigotes of Leishmania donovani by 1H NMR spectroscopy. AB - The intracellular metabolites of long-term in vitro cultured axenic amastigotes of Leishmania donovani (strain Dd8) were determined and compared with those of promastigotes and intracellular amastigotes, employing proton NMR spectroscopy. The presence of two new metabolites, i.e. betaine and beta-hydroxybutyrate were reported. Betaine was detected in all the three stages being highest in the promastigotes while beta-hydroxybutyrate could be detected only in promastigotes and axenic amastigotes. Among other metabolites, succinate and valine were found in higher quantities in intracellular amastigotes and axenic amastigotes than in promastigotes. Acetoacetate was present only in axenic and intracellular amastigotes. The comparative metabolite profile of different parasite forms reveals that axenic amastigotes seem to represent an intermediate stage between promastigotes and intracellular amastigotes in spite of their strong resemblance to intracellular amastigotes in morphology, infectivity, biochemical studies and even in the manifestation of amastigote specific A2 protein. PMID- 10465053 TI - Human serum albumin and immunoglobulin on Dracunculus medinensis. AB - Dracunculus medinensis recovered from infected humans were examined for the occurrence and localization of human serum albumin and immunoglobulins. Immunoelectrophoretic examination of homogenates prepared from adult female worms (ADGW) and first stage larvae (LVGW) showed that anti-human albumin antibodies reacted to both stages of the parasite. By direct fluorescence antibody technique, antigens resembling human albumin and human immunoglobulins (isotype IgG) were identified on the surface of adult female worms. The occurrence of host like compounds on the parasites may be an adaptation for survival of the parasites. A possible interference of these compounds should be considered in attempts to develop methods for immunological diagnosis of D. medinensis infections. PMID- 10465054 TI - Increase of intestinal schistosomiasis after praziquantel treatment in a Schistosoma haematobium and Schistosoma mansoni mixed focus. AB - The recent emergence of a mixed focus of Schistosoma haematobium-Schistosoma mansoni, in the lower delta of the Senegal river, requires adapted control programmes. A mass treatment with praziquantel was organised in April 1994 by local authorities in three villages where populations had been examined. A total of 2042 subjects participated. In Savoigne S. haematobium prevailed (53% for prevalence), in Diagambaly S. haematobium (64%) and S. mansoni (76%) were both abundant, and in Boundoum S. mansoni prevailed (53%). Therapeutic coverage (80%) was assessed on a representative sample. A cohort of 968 treated subjects were followed-up 40, 100, 200 and 300 days after treatment. Six weeks after treatment, the average of egg excretion decreased by 95% for S. haematobium, ranging from 23 to one egg(s)/10 ml at Savoigne and from 14 to one egg(s)/10 ml at Diagambal. Conversely, egg excretion only decreased by 75% for S. mansoni, from 23 to six eggs/g at Boundoum and from 69 to 16 eggs/g at Diagambal, showing evidence of the low susceptibility of S. mansoni local strain to praziquantel. Ten months after treatment, reinfections with S. haematobium remained weak at Savoigne (two eggs/10 ml) while those with S. mansoni were so high at Boundoum (24 eggs/g) that they compensated the reduction of load induced by the treatment. At Diagambal, where the two parasites were present before treatment, the disappearance of the urinary schistosomiasis after treatment concurred with a dramatic increase of intestinal schistosomiasis. S. manoni egg excretion was seven times higher than before treatment (478 eggs/g). These different effects of treatment are discussed according to the ecology of transmission in the three villages. PMID- 10465055 TI - Diagnostic value of connective tissue metabolites in Schistosoma mansoni related liver disease. AB - Reliable non-invasive markers of hepatosplenic involvement in schistosomiasis are needed for determination of morbidity levels in endemic populations and for diagnosis and follow-up of affected individuals. Serum levels of connective tissue metabolites have been investigated as fibrosis markers in various hepatic disorders, but their accuracy in the detection of hepatosplenic schistosomiasis under endemic conditions has not been fully elucidated. 206 adult inhabitants of a Tanzanian village highly endemic for schistosomiasis mansoni (prevalence 88%) underwent clinical, parasitological and sonographic work-up; sera were tested for aminoterminal procollagen III-peptide (PIIIP), carboxyterminal procollagen IV peptide (NC1) and laminin. Connective tissue marker levels did not correlate with the presence or intensity of infection. NC1 levels were significantly correlated with periportal liver fibrosis (P < 0.001), splenomegaly (P < 0.002), portal vein dilatation (P < 0.004) and the presence of portosystemic collaterals (P < 0.001); for PIIIP and laminin, none of the respective relationships was significant. Due to wide overlap of NC1 levels between individuals with normal sonography findings and those with advanced periportal fibrosis and portal hypertension, the sensitivity and positive predictive value of this markers to detect these individuals were low (< 40%), although specificity and overall accuracy in the given setting were good (80-90%). It is concluded that PIIIP and laminin are not useful as diagnostic serum markers of hepatosplenic schistosomiasis at the community level; NC1 was significantly related to various indices of hepatosplenic involvement, but its low sensitivity precludes its use as a screening tool under endemic conditions. PMID- 10465056 TI - Interleukin (IL)-4, IL-10 and IL-12 profile in serum of patients with alveolar echinococcosis. AB - Alveolar echinococcosis (AE), caused by Echinococcus multilocularis (E.m.), provokes a characteristic immune response. Based mainly on in vitro studies, Th2 dominated immunity is associated with increased susceptibility to disease, while Th1 cell activation is assumed to induce protective immunity. We investigated serum levels of interleukin (IL)4, IL-10, and IL-12 in 40 AE patients and 20 controls to assess Th1/Th2 cell activation in vivo. Significantly higher levels of IL-10 were found in AE patients (P = 0.003) than in controls, with a tendency to higher concentrations in progressive cases. In contrast, IL-4 was only measurable in a minority of patients and controls. IL-12 levels (measured with an ELISA that detects both the p35/p40 heterodimer and free p40) were comparable between AE patients and controls and showed a similar distribution pattern to IL 10 with regard to disease progression. By using an IL-12-ELISA specific for the heterodimer, only minute amounts of IL-12 were detectable in merely a minority of samples. In conclusion, our data are suggestive of Th2 dominated immune response in AE in vivo. PMID- 10465057 TI - Flow cytometric analysis of IgG reactive to parasitized red blood cell membrane antigens in Plasmodium falciparum-immune individuals. AB - Antigens exposed at the surface of Plasmodium falciparum parasitized red blood cells (pRBCs) represent potential targets for protective antibodies involved in opsonization and immune phagocytosis of pRBCs. We measured the recognition of parasitized red blood cell membrane associated antigens by IgG in the plasma of clinically immune individuals by flow cytometry and ELISA. The plasmas were selected on the basis of preexisting IgG antibodies to pRBC membrane associated recombinant proteins. In every plasma sample IgG could bind the surface of live pRBCs in flow cytometry. In addition, there was a significant correlation between the level of IgG recognition of live pRBCs and of pRBC membrane ghost proteins or major identified antigens by ELISA. Flow cytometry thus represents a technique suitable to test for the accessibility and potential functionality of IgG antibodies directed to antigens expressed by the surface of pRBCs. PMID- 10465058 TI - The cost of large-scale school health programmes which deliver anthelmintics to children in Ghana and Tanzania. The Partnership for Child Development. AB - It has been argued that the delivery of anthelmintics to school-children through existing education infrastructure can be one of the most cost-effective approaches to controlling parasitic worm infection. This paper examines the actual costs of a combination of mass and selective treatment for schistosomiasis using praziquantel and mass treatment for intestinal nematodes using albendazole, as an integral part of school health programmes reaching 80442 pupils in 577 schools in Volta Region, Ghana, and reaching 109099 pupils in 350 schools in Tanga Region, Tanzania. The analysis shows that financial delivery costs per child treated using praziquantel, which involved a dose related to body mass and a prior screening at the school level, were US$ 0.67 in Ghana and US$ 0.21 in Tanzania, while the delivery costs for albendazole, which was given as a fixed dose to all children, were US$ 0.04 in Ghana and US$ 0.03 in Tanzania. The higher unit costs in Ghana reflect the epidemiology of infection; overall, fixed costs were similar in both countries, but fewer children required treatment in Ghana. Analysis of economic costs-which includes the cost of unpaid days of labour- indicates that the financial costs are increased in Ghana by 78% and in Tanzania by 44%. It is these additional costs which are avoided by integration into an existing infrastructure. It is concluded that: the base cost of delivering a universal, standard, school-based health intervention can be as low as US$ 0.03 per child treated; that even a slight increase in the complexity of delivery can have a significant impact on the cost of intervention; and that the use of the education infrastructure does indeed offer significant savings in delivery costs. PMID- 10465059 TI - The importance of systolic control: a new paradigm for effective blood pressure management. Introduction. PMID- 10465060 TI - Influence of arterial pulse and reflective waves on systolic blood pressure and cardiac function. AB - Hypertension is a cardiovascular risk factor classically attributed to a reduction in the calibre and/or number of small arteries and arterioles resulting in increased peripheral vascular resistance. The definition of blood pressure as a product of total peripheral resistance (TPR) and cardiac output, however, does not take into account the fluctuation of blood pressure and flow during the cardiac cycle, with systolic and diastolic blood pressure representing the extremes of pulse pressure fluctuations. Diastolic blood pressure is closer to mean blood pressure (and therefore to TPR) than systolic blood pressure, and as such has been used as a marker for the diagnosis of hypertension. However, this approach has no rational basis and was challenged by the Framingham Heart Study which demonstrated that systolic rather than diastolic blood pressure is a better risk marker for stroke and coronary artery disease in subjects aged 45 years and older. This view has subsequently been confirmed by several epidemiological and interventional studies. Systolic blood pressure is closely associated with pulse pressure and is determined by the pattern of left ventricular ejection, arterial stiffness and timing of arterial wave reflections, i.e. the geometrical and viscoelastic properties of large conduit arteries. In humans, with ageing and hypertension, the arteries stiffen as a result of progressive degeneration of the arterial media, increased collagen and calcium content, and dilation and hypertrophy of large arteries and the aorta. Thus, the increase in systolic blood pressure (as a result of arterial damage) increases the fatigue of arterial walls and accelerates arterial damage, producing a self-perpetuating cycle. PMID- 10465061 TI - Elevated systolic blood pressure as a risk factor for cardiovascular and renal disease. AB - AIM: To review published literature on the relationship between systolic blood pressure and risk of cardiovascular and renal disease. DATA: identification Studies were retrieved using the MEDLINE database, bibliographies, and the authors' reference files. STUDY SELECTION: Prospective cohort studies and randomized controlled trials which were published in English-language journals. DATA EXTRACTION: All retrieved publications were reviewed by the two authors. Information on sample size, duration, study design, antihypertensive medication, participant characteristics and outcomes was abstracted for randomized controlled trials which reported reductions in systolic blood pressure during intervention. DATA SYNTHESIS: Results from several prospective cohort studies indicate that the association between systolic blood pressure and risk of coronary heart disease, stroke and end-stage renal disease is continuous, graded, and independent. Furthermore, they suggest that the association of systolic blood pressure with these outcomes is stronger than that of diastolic blood pressure. Pooling of the data available from randomized controlled trials indicates that an average reduction of 12-13 mmHg in systolic blood pressure over 4 years of follow-up is associated with a 21% reduction in coronary heart disease, a 37% reduction in stroke, a 25% reduction in total cardiovascular mortality, and a 13% reduction in all-cause mortality. CONCLUSION: Observational epidemiologic studies and randomized controlled trials have demonstrated that systolic blood pressure is an independent and strong predictor of risk of cardiovascular and renal disease. PMID- 10465062 TI - Current status of hypertensive disease treatment: results from the Evaluation and Interventions for Systolic Blood pressure Elevation: Regional and Global (EISBERG) project. AB - Conclusive evidence has shown the benefits of antihypertensive treatment The systematic review of end-point trials has indicated that for a reduction of 10-12 mmHg in systolic blood pressure or 5-6 mmHg in diastolic blood pressure, the incidence of stroke is reduced by 38% and ischaemic heart disease by 16%. Despite this, studies on the effectiveness of treatment - carried out both in specialist hypertension clinics and in the community - have all shown that patients receiving treatment for hypertension continue to be at increased risk of cardiovascular disease. Failure to control blood pressure to recommended guidelines contributes substantially to this excess risk. Some reasons for this failure are outlined below. Socioeconomic factors and lack of professional and patient compliance appear to be of considerable importance in the failure to control blood pressure. The Evaluation and Interventions for Systolic Blood pressure Elevation: Regional and Global (EISBERG) project investigated both the quantitative and qualitative aspects of the current treatment of hypertension internationally. The quantitative study indicated major shortcomings in blood pressure control, and in particular, poor control of elevated systolic pressure, which accounted for 90% of treatment failures. The qualitative research was based upon semi-structured interviews with professionals, patients and their care givers. These interviews indicated that there were misconceptions among physicians regarding the relative importance of systolic and diastolic blood pressure, the need for more aggressive treatment in the elderly, and the need for improved physician-patient interactions. If the therapeutic advances emerging from medical research are to be translated into clinical benefits, both the understanding and communication of the need for effective blood pressure control must be improved. PMID- 10465063 TI - Management of hypertension: the advent of a new angiotensin II receptor antagonist. AB - Angiotensin-converting enzyme (ACE) inhibitors and angiotensin II type 1 (AT1) receptor antagonists have received increased therapeutic recognition in the treatment of hypertension. Although the overall effects of the ACE inhibitors and AT1 receptor antagonists may seem superficially similar, there are important differences between the two classes in terms of neurohumoral activation, concomitant bradykinin potentiation, and inhibition of angiotensin II, derived not only from the classical ACE pathway, but also from alternative pathways. The AT1 receptor antagonist eprosartan has been shown to lower blood pressure effectively in hypertensive patients. When taken in the recommended dose range, 600-800 mg once daily, eprosartan is effective in patients with all grades of hypertension, regardless of age, sex or race. In several clinical studies, the blood-pressure-lowering effect of eprosartan has been shown to be at least as great as that of the ACE inhibitor enalapril. With respect to tolerability, eprosartan is superior to ACE inhibitor therapy and comparable to placebo. The pharmacological and therapeutic profiles of the expanding array of AT1 receptor antagonists differ in a number of respects. Most of these agents are biphenyl tetrazole, non-competitive antagonists, and some have active metabolites. Eprosartan differs from other AT1 receptor antagonists in that it is a non biphenyl, non-tetrazole competitive antagonist without active metabolites. Several large-scale, ongoing clinical research programmes (e.g. LIFE, SCOPE and VALUE) are expected to provide information on the extent to which AT1 receptor antagonists, in comparison with other therapeutic regimens, reduce cardiovascular morbidity and mortality in different groups of hypertensive subjects. Meanwhile, current evidence suggests that the AT1 receptor antagonists provide a new approach to the management of hypertension and that they merit a fuller assessment in other cardiovascular diseases. PMID- 10465064 TI - Pharmacological mechanism of angiotensin II receptor antagonists: implications for the treatment of elevated systolic blood pressure. AB - Systolic hypertension is a major risk factor for cardiovascular disease. The determinants of systolic blood pressure are peripheral resistance and arterial compliance. Arterial vasoconstriction, vascular growth and fluid retention, induced by the renin-angiotensin system directly or indirectly by enhancing sympathetic nervous system activity, are important factors in increasing peripheral resistance, decreasing arterial compliance and, consequently, elevating systolic blood pressure. Selective blockade of the angiotensin II type 1 (AT1) receptor represents a novel mechanism for interrupting the renin angiotensin system. This provides the additional benefit of blocking angiotensin II generated by non-angiotensin-converting-enzyme pathways without altering either bradykinin metabolism or the potential beneficial effects of AT2 receptor stimulation. Eprosartan is a potent (1.4 nmol/l) AT1 receptor antagonist that inhibits angiotensin-II-induced vascular contraction in a competitive manner. Eprosartan is effective in reducing disease progression in animal models of hypertension, heart failure, renal disease and stroke. Furthermore, eprosartan causes a large increase in arterial compliance in hypertensive rats fed high-salt and high-fat diets. Eprosartan also possesses sympathoinhibitory activity as demonstrated by an inhibition of the pressor responses induced by activation of sympathetic outflow through spinal cord stimulation in pithed rats. In contrast, other angiotensin II receptor antagonists, such as losartan, used at equivalent angiotensin II blocking activity, do not appear to alter sympathetic nervous system activity. Angiotensin II receptor antagonists, such as eprosartan, that have the ability to block both the direct effects of angiotensin II and the indirect effects mediated by enhanced sympathetic neurotransmission, may represent an important advance in the treatment of elevated systolic blood pressure. PMID- 10465065 TI - Hydroxyurea and HIV: 5 years later--from antiviral to immune-modulating effects. PMID- 10465066 TI - A soluble factor produced by macrophages mediates the neurotoxic effects of HIV-1 Tat in vitro. AB - OBJECTIVES: There is now a strong consensus that the neurotoxic properties of HIV 1 are likely to be mediated by an indirect mechanism in which neurones are damaged by infected mononuclear cells. The aim of this study was to determine the ability of HIV-1 Tat to induce neurotoxic properties in a murine macrophage cell line RAW264.7. DESIGN: Simple culture systems using dissociated neurones may not provide the appropriate microenvironment in which to observe the complex cell cell interactions that occur in the brain. We have therefore developed a more physiological model in which rat organotypic hippocampal slices are co-cultured with the murine macrophage cell line RAW264.7. Effects of Tat were studied by using a stable Tat expressing RAW264.7 cell line or by addition of recombinant Tat protein to co-cultures. METHODS: Organotypic hippocampal slices prepared from 8-10 day rat pups were grown on membrane inserts that were placed into six-well plates on which RAW264.7 cells were growing as an adherent monolayer. Cell death in the slices was assessed using propidium iodide. Specific astrocytic (glial fibrillary acidophilic protein; GFAP) and neuronal (microtubule-associated protein; MAP2) markers were visualized by immunocytochemistry. RESULTS: RAW264.7 cells that either expressed or were exposed to HIV-1 Tat protein, produced a soluble factor that caused profound degeneration in brain slice cultures involving loss of both glial cells and neurones. By contrast treatment of slice cultures with Tat in the absence of RAW264.7 cells was not neurotoxic. CONCLUSIONS: The neurotoxic properties previously attributed to HIV-1 Tat are likely to be mediated via induction of macrophage derived soluble factor(s). PMID- 10465067 TI - The CDK9-associated cyclins T1 and T2 exert opposite effects on HIV-1 Tat activity. AB - OBJECTIVES: To examine the functional interaction between HIV-1 Tat protein and the cyclin T1 and T2 proteins which, in association with cyclin dependent kinase (CDK)9, are the regulatory subunits of the TAK/P-TEFb cellular complex strictly required for Tat transactivation. DESIGN: HIV-1 long terminal repeat (LTR) reporter plasmid was co-transfected into human and rodent cells with expression vectors encoding Tat and vectors encoding the cyclins T1, T2a and T2b, respectively. METHODS: Tat-mediated transactivation of HIV-1 LTR-driven transcription was compared in the presence or absence of different cyclins T (T1, T2a and T2b), upon co-transfections into human and rodent cell lines. Protein interactions were analysed by in vitro binding assays. RESULTS: It was found that Tat function in rodent cells is enhanced by co-expression of cyclin T1 but not cyclin T2. The N-terminal region (amino acids 1-290) of cyclin T1 is sufficient for this function and for binding to Tat and CDK9. Cyclin T2 binds to CDK9 but not to Tat. Moreover, enforced expression of cyclin T2 inhibits cyclin T1 mediated enhancement of Tat in rodent cells and it represses Tat activity in human cells. CONCLUSION: Efficient Tat transactivation in rodent cells occurs in the presence of human cyclin T1 but not in the presence of cyclin T2; overexpression of cyclin T2 inhibits Tat function in both rodent and human cells. PMID- 10465068 TI - Recombinant gp160 as a therapeutic vaccine for HIV-infection: results of a large randomized, controlled trial. European Multinational IMMUNO AIDS Vaccine Study Group. AB - OBJECTIVES: The primary objective of this study was to expand the safety and immunogenicity database of recombinant gp160 as a therapeutic vaccine in the treatment of HIV-infection. Preliminary efficacy data was also sought. DESIGN: This trial was a randomized, double-blind, placebo-controlled study. Two-hundred and eight volunteers, 96 therapy-naive with CD4 cell count >500x10(6)/l (group A) and 112 with CD4 cell count of 200-500x10(6)/l (group B, 51 out of 112 on treatment with one or two nucleoside analogues), received monthly injections of rgp160 IIIB vaccine or placebo for the first 6 months of the study; booster immunizations with rgp160 MN or placebo were given at times 15, 18, and 21 months. METHODS: Safety and immunogenicity data were obtained and measurements of CD4 cell count, plasma viral RNA, and proviral DNA were performed. Clinical outcome was recorded for the 24 months of study. RESULTS: The vaccine was safe and well tolerated. Despite the induction of new rgp160-specific lymphoproliferative responses and the presence of positive delayed type hypersensitivity skin tests to rgp160 at the end of the 24 month study, no effect on the natural history of HIV infection was detected. Within 24 months, AIDS defining illnesses had occurred in 19 of the vaccinated volunteers and in 18 of the placebo recipients. Persons with higher plasma viral RNA levels and higher proviral DNA had a more rapid decline in CD4 cell count when compared to persons with lower values. Vaccine did not alter viral RNA or proviral DNA levels. CONCLUSION: There was no clinical benefit to therapeutic immunizations with rgp160, despite the induction of new lymphoproliferative responses. PMID- 10465069 TI - Human polyomavirus JC latency and reactivation status in blood of HIV-1-positive immunocompromised patients with and without progressive multifocal leukoencephalopathy. AB - BACKGROUND: Human polyomavirus JC (JCV) induces human progressive multifocal leukoencephalopathy (PML) in patients with AIDS. Peripheral blood mononuclear cells (PBMC) of HIV-1-positive immunocompetent and immunocompromised patients can harbour JCV genome, but their precise role in JCV latency or reactivation status before the onset of PML remains hypothetical. OBJECTIVES: To assess JCV latency or reactivation status in PBMC of HIV-1-positive immunocompromised patients without PML. DESIGN: A group of 82 HIV-1-positive immunocompromised patients who did not have PML were compared with 10 patients with AIDS and PML and with 69 HIV 1-positive immunocompetent patients without PML. METHODS: DNA and total RNA were extracted from PBMC. The presence of JCV DNA was demonstrated by a semi-nested polymerase chain reaction (PCR). By using primer pairs specific for an early gene,T, and a late gene, VP1, the expression of both early and late gene mRNA in PBMC could be identified using reverse transcriptase (RT) PCR. RESULTS: JCV DNA was detected by PCR in 17.4% of 69 HIV-1-positive immunocompetent patients, in 23.2% of 82 HIV-1-positive immunocompromised patients, and in 60% of 10 patients with AIDS and PML. No correlation could be drawn between the detection of JCV DNA in the PBMC and the clinical or biological status of the HIV-1-positive patients. By using RT-PCR procedures, no expression of JCV early and late mRNA in PBMC was found in any patients. CONCLUSIONS: JCV DNA is detectable in the PBMC of 20.5% of 151 HIV-1-infected patients independently of the CDC (Centers for Disease Control and Prevention) stages of the infection. Moreover, our results suggest that active replication of JCV in PBMC appears to be absent or at least a very rare event in HIV-1-positive immunocompromised patients with and without PML. PMID- 10465070 TI - Activity of non-nucleoside reverse transcriptase inhibitors against HIV-2 and SIV. AB - BACKGROUND: After the initial discovery of 1-(2-hydroxyethoxymethyl)-6 (phenylthio)thymine (HEPT) and tetrahydroimidazo[4,5,1-jk][1,4]benzodiazepin 2(1H)-one and thione (TIBO) derivatives, several other non-nucleoside reverse transcriptase (RT) inhibitors (NNRTI), including nevirapine (BI-RG-587), pyridinone derivatives (L-696,229 and L-697,661), delavirdine (U-90152), alpha anilinophenylacetamides (alpha-APA) and various other classes of NNRTI have been described. The hallmark of NNRTI has been based on their ability to interact with a specific site ('pocket') of HIV-1 RT. OBJECTIVE: To investigate whether, in addition to HIV-1, different strains of HIV-2 (ROD and EHO) and SIV (mac251, agm3 and mndGB1) are sensitive to a selection of NNRTI i.e. delavirdine, the HEPT derivative I-EBU (MKC-442), 8-chloro-TIBO (tivirapine), alpha-APA (loviride), nevirapine and the pyridinone derivative L-697,661. METHODS AND RESULTS: The NNRTI tested inhibited the replication of the different strains of HIV-2 and SIV at micromolar concentrations. The inhibitory effects of the NNRTI on HIV-2 induced cytopathicity correlated well with their inhibitory effects on HIV-2 RT activity. Drug-resistant HIV-2 (EHO) variants containing the Ser102Leu and/or Glu219Asp mutations in their RT were selected after passaging the virus in MT-4 cells in the presence of increasing concentrations of delavirdine. The EHO virus mutants were at least 20-fold less susceptible to the antiviral effects of delavirdine. Some cross-resistance, depending on the mutant strain, was observed with the other NNRTI tested (i.e. MKC-442, tivirapine, loviride and pyridinone L 697,661). CONCLUSIONS: Our data demonstrate that NNRTI are not exclusively specific for HIV-1 but are also inhibitory to different HIV-2 and SIV strains. These observations will have important implications for the development of new NNRTI with higher activity against both HIV-1 and HIV-2. Furthermore, in view of their anti-SIV activity, NNRTI could be evaluated further for their in vivo anti retrovirus efficacy in non-human primate models. PMID- 10465071 TI - Salvage therapy with regimens containing ritonavir and saquinavir in extensively pretreated HIV-infected patients. AB - OBJECTIVE: To evaluate the efficacy and toxicity of salvage regimens containing ritonavir and saquinavir in patients failing highly active antiretroviral therapy (HAART), and to correlate outcome with plasma concentrations of protease inhibitors. DESIGN: Prospective, non-randomized interventional study. SUBJECTS AND METHODS: Thirty extensively pretreated HIV-infected patients with virological failure under HAART were treated with ritonavir (400 mg twice daily) and saquinavir (600 mg twice daily) and at least one reverse transcriptase inhibitor. HIV-RNA, CD4 cell counts and plasma concentrations of protease inhibitors were determined, and patients were monitored for toxicity at monthly intervals. RESULTS: Six patients showed complete virological success (HIV-RNA <200 copies/ml at week 12) which was sustained for a median follow-up of 6.3 months. Partial virological response (decrease of HIV-RNA of >1 log10 at week 12) was achieved by a further three patients. Patients with a virological response had significantly higher CD4 cell increases than patients without virological response (mean increase at week 12: 66x10(6) cells/l versus 6x10(6) cells/l; P = 0.01). No clinical events were observed during 6 months of follow-up. Neither the use of a non-nucleoside reverse transcriptase inhibitor (NNRTI) nor the number of newly introduced drugs influenced the virological response. Plasma concentrations of protease inhibitors did not statistically differ between patients with and without success. Toxicity included gastrointestinal disturbances, lipid abnormalities and liver dysfunction. CONCLUSIONS: In extensively pretreated patients, salvage regimens containing ritonavir and saquinavir had only limited and short-term anti-HIV activity and were associated with substantial toxicity. Plasma concentrations of saquinavir were not predictive for virological response. PMID- 10465072 TI - Cerebrospinal fluid HIV-1 RNA levels in asymptomatic patients with early stage chronic HIV-1 infection: support for the hypothesis of local virus replication. AB - OBJECTIVE: To assess HIV-1 RNA levels in cerebrospinal fluid (CSF) and their potential correlation with plasma viral load and central nervous system (CNS) HIV 1 infection markers in stable asymptomatic patients with a CD4 T cell count >500x10(6) cells/l. PATIENTS AND METHODS: Consecutive patients screened for two trials were eligible for lumbar puncture assessment. At day 0, simultaneous samples of CSF and plasma were obtained and levels of total proteins, albumin, IgG, antibodies against HIV-1 p24 antigen, HIV-1 RNA (using the polymerase chain technique) and white cells were measured. RESULTS: The integrity of the blood brain barrier was preserved (albumin index > or =7) in 59 out of 70 patients (84%). Intrathecal production of antibodies against HIV-1 p24 antigen was demonstrated in 55 out of 70 individuals (78%). Viral load in CSF was significantly lower than plasma values (3.13+/-0.95 versus 4.53+/-0.53, P = 0.0001). HIV-1 RNA was not detected in CSF in only three of the 70 patients (4%). Overall, there was a significant correlation between plasma and CSF HIV-1 RNA levels (r = 0.43, P = 0.0001); however, in 29 patients (41%) there were significant differences (>1.5 log10 copies/ml) between the viral loads in plasma and CSF. In the multivariate analysis, a high level of protein and white cells in CSF, but not the HIV-1 RNA plasma level, were factors independently associated with a higher level of HIV-1 RNA in CSF (P = 0.0001). CONCLUSIONS: HIV-1 RNA can be detected almost always in CSF of asymptomatic patients in early stages of HIV 1 infection including those with a preserved integrity of the blood-brain barrier. The important discrepancies between plasma and CSF viral load, and the independent association between CSF abnormalities and CSF viral load, support the hypothesis of local production of HIV-1. PMID- 10465073 TI - Incidence and risk factors for developing cytomegalovirus retinitis in HIV infected patients receiving protease inhibitor therapy. Spanish CMV-AIDS Study Group. AB - OBJECTIVE: To assess the incidence and risk factors for cytomegalovirus (CMV) retinitis in HIV-infected patients who initiated protease inhibitor-containing antiretroviral therapy. DESIGN AND SETTING: Prospective, multicentre study. PATIENTS: A cohort of 172 HIV-infected patients with a CD4 cell count below 100x10(6) cells/l at the time of protease inhibitor introduction. MAIN OUTCOME MEASURES: Confirmed CMV retinitis and mortality, according to CD4 cell count, HIV load, and CMV viraemia. RESULTS: The cumulative incidence of CMV retinitis was 5% at 1 year and 6% at 2 years. Only a positive CMV polymerase chain reaction (PCR) test at therapy initiation was significantly associated with the development of disease (relative hazard, 4.41; 95% confidence interval, 2.12-8.93; P<0.00001). The 12-month Kaplan-Meier CMV retinitis event rate was 38% in patients who were CMV PCR-positive compared with 2% in those who were CMV PCR-negative (P<0.001). Mean CMV load was significantly higher in those individuals who went on to develop CMV retinitis (3700 versus 384 copies/ml, P = 0.002). Only 2% of patients remained CMV PCR-positive after 3 months of protease inhibitor therapy, and CMV viraemia was not associated with a worse therapy response or shorter survival. Transient CMV positivity without a higher risk of disease was observed in 7% of patients at the first month on therapy. CONCLUSIONS: Protease inhibitor containing antiretroviral therapy significantly reduces the incidence of CMV viraemia and disease. Although a positive CMV PCR test identifies those patients on therapy at highest risk of CMV retinitis, it is not associated with an increased risk of death or a worse response to protease inhibitor therapy. PMID- 10465074 TI - Sampling lymphoid tissue cells by ultrasound-guided fine needle aspiration of lymph nodes in HIV-infected patients. Swiss HIV Cohort Study. AB - OBJECTIVE: To establish the feasibility of using ultrasound-guided lymph node needle aspiration as a means to obtain lymphoid tissue cells for the determination of a series of immunologic and virologic measures in HIV-infected patients. DESIGN: First, a comparison of the characteristics of cell populations obtained by simultaneous needle aspiration and standard excisional biopsy in six patients. Second, use of lymph node needle aspiration to assess longitudinally T cell subset changes in patients initiating highly effective antiretroviral treatment. METHODS: T-cell subsets (CD4 and CD8) and percentage Ki67+ cycling T cells were measured in lymph node cell populations harvested by ultrasound-guided aspiration or standard biopsy by flow cytometry. Cellular RNA content was assessed by a modification of the Roche Amplicor HIV-1 Monitor test. RESULTS: CD4 and CD8 T-cell percentage and HIV RNA cell content of lymph node cell suspensions obtained from the simultaneous performance of ultrasound-guided needle aspiration and excisional biopsy in the same patients were correlated (n = 6). Among the 87 aspiration sessions reported here, mononuclear cell suspensions were obtained in 100% of the sessions, in numbers ranging between 4x10(4) to 6.7x10(6) cells (median: 7x10(5)). This limited number of cells did not allow to perform all type of analyses in all patients. By prioritizing the cells for the determination of T cell subsets and proliferation rate, this approach was instrumental for demonstrating the normalization of the T-cell subset ratio and the kinetic of normalization of proliferating rates of CD4 and CD8 T cells, as well as the decrease in HIV-1 viral load in the lymph node following HAART initiation. CONCLUSION: Ultrasound-guided aspiration appears to be a non-invasive and ad libitum, safe and repeatable procedure for the longitudinal monitoring of changes in lymph nodes. PMID- 10465075 TI - Phase 1 trial of nonoxynol-9 film among sex workers in South Africa. AB - OBJECTIVES: To assess the acceptability and safety of a vaginal nonoxynol-9 film in a group of sex workers at a truck stop in South Africa. DESIGN: A randomized double-blinded crossover trial was conducted between April 1995 and July 1995. INTERVENTION: Seventy-two mg nonoxynol-9 film and an identical glycerine placebo film. METHODS: Following informed consent, each study participant was randomly assigned the designated pre-coded film for 1 month. The second month was a film free washout period and the participants used the alternate film in the third month. Besides measuring behavioural and clinical outcomes, colposcopy examination for genital lesions, serology and microbiology investigations for sexually transmitted diseases and semi-quantitative PCR for vaginal HIV load estimates were performed. RESULTS: Twenty women participated in the study. The women reported, on average, 19 sexual encounters per week. Vaginal intercourse was protected 25% of the time by condoms. On average, 11 vaginal films, either nonoxynol-9 or placebos were inserted per week. There were no statistically significant differences between the two treatment groups for genital lesions (P = 0.29), reported side effects (P = 0.73), and viral load (P = 0.9). However, the proportions of clinically detected genital lesions (six out of eight versus two out of eight) and self-reported side-effects (five out of eight versus three out of eight) were higher in the nonoxynol-9 group when compared with the placebo group. Incident sexually transmitted diseases occurred more frequently in the placebo group. An increased viral load was associated with the development of a genital lesion (relative risk, 6.0; 95% confidence interval, 0.81-44.4). CONCLUSIONS: The 72 mg film formulation of nonoxynol-9 was an acceptable product for use in this population of sex workers. Although no statistically significant differences in adverse outcomes were detected, clinically there appeared to be an increase in minor lesions and self-reported side-effects with nonoxynol-9 and less protection against sexually transmitted diseases with the placebo. Furthermore, HIV shedding was correlated with the presence of incident vaginal or cervical lesions. This brings into question the potential narrow margin of safety for this product; additional Phase 2 studies are therefore required. PMID- 10465076 TI - Randomized trial testing the effect of vitamin A supplementation on pregnancy outcomes and early mother-to-child HIV-1 transmission in Durban, South Africa. South African Vitamin A Study Group. AB - OBJECTIVE: Poor vitamin A status has been associated with a higher risk for mother-to-child transmission of HIV-1 and there is contradictory evidence on the impact of vitamin A on perinatal outcome. We therefore assessed the effect of vitamin A supplementation to mothers on birth outcome and mother-to-child transmission of HIV-1. DESIGN AND METHODS: In Durban, South Africa 728 pregnant HIV infected women received either vitamin A (368) or placebo (360) in a randomized, double-blind trial. The vitamin A treatment consisted of a daily dose of 5000 IU retinyl palmitate and 30 mg beta-carotene during the third trimester of pregnancy and 200000 IU retinyl palmitate at delivery. HIV infection results were available on 632 children who were included in the Kaplan-Meier transmission analysis. Results are reported on mother-to-child transmission rates up to 3 months of age. RESULTS: There was no difference in the risk of HIV infection by 3 months of age between the vitamin A [20.3%; 95% confidence interval (CI), 15.7 24.9] and placebo groups (22.3%; 95% CI, 17.5-27.1), nor were there differences in foetal or infant mortality rates between the two groups. Women receiving vitamin A supplement were, however, less likely to have a preterm delivery (11.4% in the vitamin A and 17.4% in the placebo group; P = 0.03) and among the 80 preterm deliveries, those assigned to the vitamin A group were less likely to be infected (17.9%; 95% CI, 3.5-32.2) than those assigned to the placebo group (33.8%; 95% CI, 19.8-47.8). CONCLUSION: Vitamin A supplementation, a low-cost intervention, does not appear to be effective in reducing overall mother-to-child transmission of HIV; however, its potential for reducing the incidence of preterm births, and the risk of mother-to-child transmission of HIV in these infants needs further investigation. PMID- 10465077 TI - Gay men report high rates of unprotected anal sex with partners of unknown or discordant HIV status. AB - OBJECTIVE: To examine patterns and factors that correlate with unprotected anal intercourse (UAI) practices among San Francisco gay men, including UAI with partners of unknown or different HIV antibody status. DESIGN: A longitudinal cohort recruited for the San Francisco Young Men's Health Study in 1992; re assessed annually. PARTICIPANTS AND METHODS: A sample of 510 unmarried gay men who were 18 to 29 years at baseline were originally recruited as part of a larger population and referral-based sample. Subjects participated in four consecutive waves of data collection. RESULTS: The prevalence of reported unprotected anal intercourse (UAI) increased from 37% to 50% between 1993-1994 and 1996-1997. Almost half of all men who reported UAI in 1996-1997 indicated that it occurred with a partner of unknown or discordant HIV antibody status. This high-risk practice correlated with greater numbers of male sex partners, use of nitrite inhalants, sex in commercial sex environments, perceived difficulty controlling sexual risk-taking, and negative emotional reactions following UAI. CONCLUSIONS: These data on increasing rates of sexual risk-taking further confirm trends in sexual behavior previously suggested by rising rates of rectal gonorrhea in this population. Additional and sustained prevention efforts are urgently needed in light of the very high background rates of HIV infection found among gay men in San Francisco. PMID- 10465078 TI - Mortality in young adults in England and Wales: the impact of the HIV epidemic. AB - OBJECTIVE: To quantify the contribution of the HIV epidemic to premature mortality in England and Wales 1985-1996. DESIGN: Surveillance of deaths in HIV infected individuals and causes of death from death certificates. MAIN OUTCOME MEASURES: Time trends in age-specific mortality rates among 15-44 year olds and years of potential life lost (YPLL) to age 65 associated with HIV infection and other important causes of death in young adults. RESULTS: The crude age-specific mortality rates for all causes of death in the 15-44 year age band remained fairly constant between 1985 and 1996: in other age bands a decrease was seen. Deaths from both suicide and HIV increased in men aged 15-44 years. Although suicide accounted for a greater number of deaths throughout the period investigated, the largest proportional and absolute increase was seen for deaths in HIV-infected people. By 1996, the contribution of HIV to YPLL to age 65 varied from less than 0.5% in most rural localities to 20% of total YPLL in one London health authority. CONCLUSIONS: While part of the adverse trend in mortality in younger adults since 1985 was attributable to suicide, most resulted from HIV infection. The impact of HIV infection on mortality was greatest in London. PMID- 10465079 TI - Relapse and mortality among HIV-infected and uninfected patients with tuberculosis successfully treated with twice weekly directly observed therapy in rural South Africa. AB - OBJECTIVE: To determine post-treatment relapse and mortality rates among HIV infected and uninfected patients with tuberculosis treated with a twice-weekly drug regimen under direct observation (DOT). SETTING: Hlabisa, South Africa. PATIENTS: A group of 403 patients with tuberculosis (53% HIV infected) cured following treatment with isoniazid (H), rifampicin (R), pyrazinamide (Z) and ethambutol (E) given in hospital (median 17 days), followed by HRZE twice weekly to 2 months and HR twice weekly to 6 months in the community under DOT. METHODS: Relapses were identified through hospital readmission and 6-monthly home visits. Relapse (culture for Mycobacterium tuberculosis) and mortality given as rates per 100 person-years observation (PYO) stratified by HIV status and history of previous tuberculosis treatment. RESULTS: Mean (SD) post-treatment follow-up was 1.2 (0.4) years (total PYO = 499); 78 patients (19%) left the area, 58 (14%) died, 248 (62%) remained well and 19 (5%) relapsed. Relapse rates in HIV-infected and uninfected patients were 3.9 [95% confidence interval (CI) 1.5-6.3] and 3.6 (95% CI 1.1-6.1) per 100 PYO (P = 0.7). Probability of relapse at 18 months was estimated as 5% in each group. Mortality was four-fold higher among HIV-infected patients (17.8 and 4.4 deaths per 100 PYO for HIV-infected and uninfected patients, respectively; P<0.0001). Probability of survival at 24 months was estimated as 59% and 81%, respectively. We observed no increase in relapse or mortality among previously treated patients compared with new patients. A positive smear at 2 months did not predict relapse or mortality. CONCLUSION: Relapse rates are acceptably low following successful DOT with a twice weekly rifampicin-containing regimen, irrespective of HIV status and previous treatment history. Mortality is substantially increased among HIV-infected patients even following successful DOT and this requires further attention. PMID- 10465080 TI - Tuberculosis preventive therapy for HIV-infected people in sub-Saharan Africa is cost-effective. AB - OBJECTIVE: Since antiretroviral therapy is largely unavailable to HIV-infected patients in developing countries and recent clinical trials have shown that tuberculosis (TB) preventive therapy can reduce TB and HIV-associated morbidity and mortality, we studied the effectiveness and cost-effectiveness of TB preventive therapy for HIV-infected persons in sub-Saharan Africa. METHODS: A Markov model that used results of clinical trials of TB preventive therapy in sub Saharan Africa and literature-derived medical care costs was used to evaluate three preventive therapy regimens in HIV-infected, tuberculin-positive patients in Uganda: (1) daily isoniazid (INH) for 6 months, (2) daily INH and rifampin (RIF) for 3 months, and (3) twice-weekly RIF and pyrazinamide (PZA) for 2 months. RESULTS: All three regimens extend life expectancy and reduce the number of TB cases. When only medical care costs are considered, all three preventive therapy regimens cost more than not providing preventive therapy to extend life and prevent active tuberculosis. When medical care and social costs are considered together, 6-months of daily INH treatment will save money relative to no preventive therapy and when the costs associated with treating secondary infections are included, all three preventive therapy regimens are less expensive than no preventive therapy. With the inclusion of secondary infection costs, 6 months of daily INH results in savings of $24.16 per person. CONCLUSIONS: TB preventive therapy taken by HIV-infected tuberculin reactors in sub-Saharan Africa results in extended life-expectancy, reduction of the incidence of TB and monetary savings in medical care and social costs. TB control policy in sub Saharan Africa should include preventive therapy. PMID- 10465081 TI - A psychological epidemiology of people seeking HIV/AIDS counselling in Kenya: an approach for improving counsellor training. AB - OBJECTIVE: This study develops a typology of psychological problems reported during HIV/AIDS counselling. This typology provides a framework for training paraprofessional counsellors (PPCs) in East Africa. DESIGN: Study participants included 307 Kenyans tested for HIV at any of six clinics in Nairobi specialising in STDs, tuberculosis and other infectious diseases. Pre-test, post-test, and follow-up counselling was provided by 16 PPCs who are themselves HIV-positive. Data consisted of demographic, physical and psychological information reported by 168 clients who sought follow-up counselling. METHODS: Counselling data were coded using an ipsative method; a unique code was assigned to every distinct topic. Factor analysis with a Varimax rotation reduced the original psychological variables into logical groupings. Multivariate analysis examined the relationship of factors and demographic characteristics. RESULTS: Clients reported 1-10 physical and 1-23 psychological complaints in a single session. Sixty-five percent of female clients reported > or = eight psychological problems; 49% of males reported > or = eight psychological problems. Factor analyses allowed the 109 reported psychological events to be assigned to 15 categories of problems. Multivariate analyses explained little of the variance in the relationship between each client's demographic profile and the psychological factors. CONCLUSIONS: Training for PPCs should be relevant to problems encountered during counselling. Results indicate that PPCs can expect clients to present one or more of the 15 factors during counselling. Demographic characteristics explained small amounts of variance in the distribution of factor scores. The 15 factors produced in this study, although descriptive and preliminary, could form the basis of a training curriculum for HIV PPCs. PMID- 10465082 TI - Costs and benefits to the mother of antenatal HIV testing: estimates from simulation modelling. AB - OBJECTIVE: To assess the health service costs and benefits for the woman of an earlier HIV diagnosis as a result of antenatal HIV testing. DESIGN: A model of maternal disease progression was developed based on the rate of decline in CD4 cell counts and applied to two matched simulated cohorts of women with identical initial CD4 cell levels and decline rates but whose HIV diagnosis occurred at different times as a result of antenatal HIV testing. UK data on CD4 cell count at HIV diagnosis and annual health service costs of care excluding antiretroviral therapy (ART) incurred at defined states of CD4 cell count were taken from published UK data. Costs of triple ART were added and effectiveness modelled by retarding the rate of CD4 cell count decline. Discounting costs at 6% and life years at 2% per year, the additional costs per life-year gained by screening were calculated. Uncertainty was explored using sensitivity analysis. RESULTS: Costs per life-year gained by antenatal diagnosis of women were pound sterling 51258 ($76887) assuming untested women were diagnosed a median of 20.4 months later than tested women, ART was initiated at a CD4 cell count of 350x10(6) cells/l and ART efficacy retarded decline in CD4 cell counts by 40% for life. Sensitivity analyses showed results were most sensitive to the assumed efficacy of lifetime ART and time assumed to HIV diagnosis for women not tested in pregnancy. CONCLUSION: This model provides a way of estimating the additional costs and benefits of future care for the woman resulting from an earlier HIV diagnosis through antenatal testing. These should be included with the paediatric costs averted and life-years gained from interventions to reduce mother-to-child transmission in order to evaluate the cost-effectiveness of antenatal screening in different populations and settings. PMID- 10465083 TI - HIV counseling in sub-Saharan Africa. PMID- 10465084 TI - Tuberculosis preventive therapy in people living with HIV in sub-Saharan Africa. PMID- 10465085 TI - HLA-unrestricted killing of HIV-1 gag protein-expressing CD4 T cells by gag specific CD8 cytotoxic T cells. PMID- 10465086 TI - Characterization of a third CCR5 amplicon from CCR5-delta32-heterozygous HIV-1 infected individuals. PMID- 10465087 TI - Reversible zidovudine-induced pure red cell aplasia. PMID- 10465088 TI - Quality of life in asymptomatic patients with early HIV infection initiating antiretroviral therapy. Swiss HIV Cohort Study. PMID- 10465089 TI - Treatment of painful distal sensory polyneuropathy in HIV-infected patients with a topical agent: results of an open-label trial of 5% lidocaine gel. PMID- 10465090 TI - Serum thrombopoietin levels correlate with disease progression of AIDS. PMID- 10465091 TI - Salvage therapy in HIV-positive patients with a previous therapeutic experience with protease inhibitors. Members of the Master Group. PMID- 10465092 TI - Hydroxyurea-induced neurotoxicity in HIV disease. PMID- 10465093 TI - Changes in plasma HIV-RNA and CD4 lymphocyte counts in patients receiving highly active antiretroviral therapy. PMID- 10465094 TI - The frequency of anonymous HIV testing in the United States: data from a national survey. PMID- 10465095 TI - Sentinel surveillance for HIV infection and risk behavior among adolescents entering juvenile detention in San Francisco: 1990-1995. PMID- 10465096 TI - Adjustment of antenatal clinic HIV surveillance data for HIV-associated differences in fertility. PMID- 10465097 TI - Sexual behavioural study of commercial sex workers and their clients in Cambodia. Japan-Cambodia Collaborating Research Group. PMID- 10465098 TI - Changes in viral load during acute respiratory infections in HIV-infected patients. PMID- 10465099 TI - Mixed chimerism: preclinical studies and clinical applications. AB - Traditional approaches to allogeneic stem cell transplantation have relied on the use of toxic high-dose conditioning therapy to achieve allogeneic engraftment and control of underlying disease. Preclinical observations have shown that, for engraftment purposes, conditioning regimens can be reduced in intensity, resulting in reduced treatment toxicities. In preclinical canine studies, the use of potent pre- and postgrafting immunosuppression allowed for reduction in conditioning regimens and development of stable mixed chimerism. If these newer approaches using attenuated conditioning regimens can be successfully applied to human transplantation, an improved safety profile will allow potentially curative treatment of patients not currently offered such therapy. Mixed chimerism per se could prove curative of disease manifestation for various nonmalignant disturbances of the hematopoietic and immune systems. For patients with malignancy, infusion of additional donor lymphocytes may be needed to effectively treat underlying disease. PMID- 10465100 TI - Manganese [correction of Magnesium] superoxide dismutase (MnSOD) plasmid/liposome pulmonary radioprotective gene therapy: modulation of irradiation-induced mRNA for IL-I, TNF-alpha, and TGF-beta correlates with delay of organizing alveolitis/fibrosis. AB - Radiation pneumonitis remains a critical dose-limiting toxicity of total body irradiation (TBI) for use in bone marrow transplantation. The acute and chronic phases of radiation damage in the mouse lung have been shown to correlate with mouse strain genotype and are dependent on fraction size, total dose, and total lung volume. Our prior studies demonstrated effective prevention of irradiation induced lung damage and improved survival in C57BL/6J mice by MnSOD plasmid/liposome gene therapy. In the present studies, we investigated the kinetics of irradiation-induced upregulation of mRNA for acute phase cytokines interleukin (IL)-1 and tumor necrosis factor (TNF)-alpha, and fibrosis-associated transforming growth factor (TGF)-beta and isoforms (TGF-beta1, TGF-beta2 and TGF beta3) in 2000 cGy whole-lung irradiated C57BL/6J mice, a strain known to develop dose and volume-dependent organizing alveolitis/fibrosis. The results demonstrate increase in mRNA for IL-1 between days 1 and 14 after irradiation with return to baseline levels out to 120 days. TNF-alpha mRNA levels were not initially elevated but increased between 80 and 100 days and then decreased by 120 days. The mRNA levels for TGF-beta1 demonstrated an initial increase within the first 14 days after total lung irradiation with a decrease to baseline levels out to 100 days. Then, in striking contrast to the other two cytokines, an increase in TGF-beta2 mRNA occurred at around 120 days and correlated with the detection of organizing alveolitis/radiation fibrosis and mortality. These results are consistent with a two-phase mechanism in the molecular pathology of irradiation lung injury, in which IL-1 cytokine mRNA levels correlated with the acute pneumonitis phase and delayed elevation of TNF-alpha (80-100 days), TGF-beta1 (100 days), and TGF-beta2 (120 days) were associated with the fibrosis phase. Insight into the cell-specific and tissue-specific molecular mechanisms of ionizing irradiation induction of mRNA for pulmonary cytokines may provide new strategies for treatment of radiation pneumonitis in TBI patients. PMID- 10465101 TI - Retroviral vector-mediated gene transfer into umbilical cord blood-derived megakaryocyte and platelet progenitors. AB - Optimizing platelet engraftment following hematopoietic stem cell transplantation is essential for minimizing transplant-related morbidity, particularly following umbilical cord blood transplantation (UCBT), where platelet engraftment frequently takes >60 days. One strategy for optimizing platelet engraftment following UCBT is to study or alter the genetic program of megakaryocyte/platelet (Mk/plt) progenitors. Retroviral vector gene transfer has previously proven useful for studying the biology of hematopoietic stem cells; however, procedures for transducing UCB cells of the Mk/plt lineage with retroviral vectors have not been described. We report here that Mk/plt progenitors generated from UCB progenitors can be efficiently transduced with retroviral vectors. Transduced Mk/plt cells were identified and quantitated by expression of a vector transgene encoding a truncated version of the human nerve growth factor receptor (NGFR). Vector-mediated NGFR expression could be readily detected in Mk/plt progenitors defined by immunophenotypic, morphologic, and functional criteria. In addition, NGFR expression persisted in mature anucleate platelets generated from the transduced Mk/plt progenitors. These methods may be useful for introducing genetic elements into Mk/plt progenitors to study various aspects of platelet development and biology and for marking ex vivo expanded Mk/plt progenitors to determine their contribution to engraftment. PMID- 10465102 TI - Graft-facilitating doses of ex vivo activated gammadelta T cells do not cause lethal murine graft-vs.-host disease. AB - The purpose of this study was to examine the ability of gamma(delta) T cells to cause graft-vs.-host disease (GVHD) after allogeneic bone marrow transplantation (BMT) and to determine whether these cells offered any therapeutic advantages relative to alphabeta T cells. Due to the paucity of naive gamma(delta) T cells in mice and humans, gamma(delta), T cells (obtained from alpha(beta) T cell deficient murine donors) were ex vivo activated and expanded in interleukin (IL) 2 so as to achieve sufficient cell numbers and to serve as a more clinically feasible strategy. After transplantation into lethally irradiated hosts, donor gamma(delta) T cells were detected in target organs of GVHD such as the spleen and intestines 2 weeks after BMT and constituted the primary T cell subpopulation. Large doses (150 x 10(6)) of activated gamma(delta) T cells, which we have previously shown capable of facilitating engraftment in MHC-disparate recipients, failed to cause fatal GVHD in lethally irradiated recipients of MHC incompatible donor marrow grafts (C57BL/6 [H-2b]-->B10.BR [H-2k] and C57BL/6 [H 2b]-B6D2F1[H-2b/d]). The absence of GVHD was confirmed by histologic analysis of target organs, splenic B cell reconstitution, and appropriate negative selection in the thymus, that were all comparable to those observed in mice transplanted with T cell-depleted BM only. While early splenic reconstitution was attributable to donor gamma(delta) T cells, analysis of durably engrafted chimeras 2 months posttransplant revealed that the vast majority of donor splenic T cells expressed the alpha(beta) T cell receptor. The results of secondary adoptive transfer assays showed that these cells were tolerant of recipient alloantigens in vivo, demonstrating that gamma(delta) T cells did not prevent the subsequent development of donor anti-host tolerance in BM-derived alpha(beta) T cells. When comparatively evaluated, the minimal number of naive alpha(beta) T cells necessary for donor engraftment caused significantly more fatal GVHD than the corresponding minimal dose of activated gamma(delta) T cells and thus had a superior therapeutic index. These studies indicate that doses of activated gamma(delta) T cells that are able to promote alloengraftment do not cause lethal GVHD in mice transplanted with MHC-incompatible marrow grafts. PMID- 10465103 TI - Antileukemic effect of interleukin-2-transduced murine bone marrow after autologous transplantation. AB - Myeloablative chemotherapy or radiation therapy supported by autologous stem cell transplantation (SCT) for the treatment of hematologic malignancies such as acute leukemia, lymphoma, and myeloma is associated with high rates of relapse. The reasons for this are 1) autologous transplantation lacks the in vivo graft-vs. tumor (GVT) effect associated with allogeneic SCT, which is effective in controlling or eliminating residual malignant cells remaining in the body after high-dose therapy, and 2) contaminating malignant cells in the autologous graft are reinfused into the body. Some researchers have attempted to administer immunomodulatory cytokines to simulate a GVT effect, and although this has shown some efficacy, it has several disadvantages. These include high toxicity associated with systemic administration, a short in vivo half-life, and insufficient levels reaching the site of residual disease. As an alternative, we investigated whether delivery of the cytokine interleukin (IL)-2 to the bone marrow can exert an antileukemic effect while avoiding the problems associated with systemic administration. We describe the delivery of IL-2 to the bone marrow by transplantation of syngeneic bone marrow, retrovirally transduced with the gene for IL-2, into lethally irradiated mice. We were able to efficiently transduce murine bone marrow with the IL-2 gene without adversely affecting clonogenic output from hematopoietic progenitors, and we were able to achieve expression of the transgene in transplanted animals. However, IL-2 transduction inhibited hematopoietic reconstitution in lethally irradiated mice. Marrow transduced with high-titer, high-expressing IL-2 retrovirus failed to engraft, and a low-titer, low-expressing IL-2 retrovirus also demonstrated reduced engraftment, although engraftment was sufficient to support survival of transplanted mice. Long-term, low-level expression of the IL-2 transgene was detectable in these mice and was effective in exerting an antileukemic effect. Mice transplanted with control marrow and challenged with leukemic cells suffered 100% mortality within 70 days, whereas mice transplanted with IL-2-transduced marrow exhibited 50% survival over the 175-day duration of this study. The work shows that delivery of immunomodulatory cytokines to the bone marrow can be achieved by transplantation of genetically modified hematopoietic cells. Furthermore, low-level IL-2 expression can exert an antileukemic effect. These data suggest that this may be an effective immunotherapeutic strategy to reduce relapse after autologous transplantation, but the selection and expression of the cytokine must be carefully considered to minimize adverse effects on hematopoiesis. PMID- 10465104 TI - Marrow transplantation from unrelated donors for patients with severe aplastic anemia who have failed immunosuppressive therapy. AB - Allogeneic marrow transplantation offers curative therapy for patients with aplastic anemia. We analyzed retrospective results in 141 patients with severe aplastic anemia who received transplants between 1988 and 1995 from an unrelated volunteer donor identified through the National Marrow Donor Program (NMDP). All patients had failed one or more courses of immunosuppressive therapy. Of the patients, 121 (86%) received a radiation-containing conditioning regimen, and 20 (14%) were given chemotherapy only. Based on serologic human leukocyte antigen (HLA) typing (class I and II), 105 patients (74%) received HLA-matched marrow, and 36 (26%) received marrow mismatched for at least one HLA-A, -B, or -DR antigen. Allele-level (molecular) typing for HLA-DRB1 was available in 108 donor recipient pairs; 77 patients received DRB -matched and 31 DRB1-mismatched transplants. All but 13% of patients were given a cyclosporine-containing regimen for graft-vs.-host disease (GVHD) prophylaxis, and 45 patients (32%) received marrow that was T cell-depleted. Among 131 evaluable patients, 116 (89%) achieved sustained engraftment and 15 (11%) did not. Among patients with engraftment, acute GVHD of grades II-IV developed in 60 patients (52%) and extensive chronic GVHD in 24 patients at risk (31%). Currently, 51 patients (36%) are surviving at 11-94 months (median 36) after transplantation. All but five have Karnofsky scores > or =80. Patients who received a serologically matched transplant fared somewhat better than did patients given a serologically mismatched transplant p = 0.03). Patients with donors matched by both serology and allele-level DRB1 typing had significantly better survival than DRB1-mismatched patients with 56 vs. 15% surviving at 3 years p = 0.001). Outcome in patients transplanted within 3 years of diagnosis was superior to that among patients transplanted with greater delay. Major causes of death were graft failure, GVHD, and infections. These data suggest that unrelated marrow transplantation offers successful therapy for a proportion of patients who have failed immunosuppressive therapy. PMID- 10465105 TI - Long-term follow-up of patients who achieved complete remission after donor leukocyte infusions. AB - Donor leukocyte infusions (DLI) can induce a direct graft-vs-leukemia (GVL) reaction and restore complete remission for patients who relapse after allogeneic bone marrow transplantation (BMT). A critical and unanswered concern is the long term safety and durability of DLI. To determine remission duration, long-term toxicity, and survival after DLI-induced remissions, we identified 73 patients who achieved complete remission after DLI. Follow-up information was obtained for 66 of the 73 patients, including 39 patients with chronic myelogenous leukemia (CML) and 27 patients with other diseases. Median follow-up for all patients was 32 months; the probability of survival at 1, 2, and 3 years was 83% (95% confidence interval [CI] 74-92), 71% (60-83), and 61% (49-74), respectively. For CML, survival probability at 1, 2, and 3 years was 87% (76-98), 76% (62-90), and 73% (58-88). For other diseases, survival probability at 1 and 2 years is 77% (61 93) and 65% (46-84). Five of 39 patients with CML relapsed, and 11 of 27 patients with other diseases relapsed. Treatment-related toxicity accounted for 10 deaths. Extended follow-up shows that DLI-induced remissions are durable, especially for patients with CML. Late relapses still occur, however, and toxicity remains significant. Continued follow-up will best define the long-term GVL effects of DLI, especially for diseases other than CML. PMID- 10465106 TI - Autologous bone marrow transplantation after histologic transformation of indolent B cell malignancies. AB - The role of high-dose therapy and autologous stem cell transplantation in diffuse large B cell lymphoma (DLBCL) after transformation is controversial. We have retrospectively analyzed patients with chemosensitive disease and a history of follicular lymphoma or chronic lymphocytic leukemia/small lymphocytic lymphoma who underwent high-dose chemoradiotherapy and bone marrow transplantation (BMT) with anti-B cell monoclonal antibody-purged autologous marrow for DLBCL. Between December 1982 and August 1997, 27 patients underwent autologous BMT using a uniform ablative regimen with cyclophosphamide, total-body irradiation, and bone marrow purging. All patients received multiple chemotherapy regimens before autologous BMT. At bone marrow (BM) harvest, only 44% of patients were in complete remission, and overt BM infiltration was present in 37%. After cyclophosphamide and total-body irradiation, no treatment-related deaths were seen. Eleven of the 27 patients relapsed, and four patients developed myelodysplasia/acute myelogenous leukemia. In seven patients in whom pathologic studies were available after relapse, the histology remained DLBCL. Twelve patients remained alive and in complete remission with a median follow-up of 36 months (range 10-132). The disease-free survival and overall survival are estimated to be 46% (90% confidence interval 28-64) and 58% (40-76) at 5 years, respectively. Patients whose disease underwent histologic transformation within 18 months of their initial diagnosis of indolent lymphoma had significantly better overall survival. Selected patients with histologic transformation, particularly those whose transformation occurs early in the course of their disease and who remain chemosensitive, may experience prolonged survival after autoBMT. PMID- 10465108 TI - Prevalence, age of onset, and natural history of thyroid disease in Pendred syndrome. AB - BACKGROUND: We have sought to establish the prevalence of goitre within a Pendred syndrome (PS) cohort and to document the course of thyroid disease in this patient group. As part of a genetic study of PS we have assessed 57 subjects by perchlorate discharge test and in 52 (M 21, F 31, age range 9-54 years) a discharge of radioiodide of >10% was observed. RESULTS: Goitre was present in 43 (83%) of the cohort (28 F, 15 M), generally developing after the age of 10 years, 56% remained euthyroid (age range 9-37 years), and 19 patients (44%) had objective evidence of hypothyroidism, all of whom had goitre. CONCLUSIONS: In summary, thyroid dysfunction in PS is variable and inclusion of goitre as a diagnostic requirement will maintain significant underascertainment. The recent identification of the genetic defect underlying PS is likely to provide an important diagnostic aid in the identification of this disorder and this communication should assist clinicians in identifying deaf patients who ought to be considered for this investigation. PMID- 10465107 TI - Genetics of bipolar disorder. AB - Bipolar disorder (also known as manic depressive illness) is a complex genetic disorder in which the core feature is pathological disturbance in mood (affect) ranging from extreme elation, or mania, to severe depression usually accompanied by disturbances in thinking and behaviour. The lifetime prevalence of 1% is similar in males and females and family, twin, and adoption studies provide robust evidence for a major genetic contribution to risk. There are methodological impediments to precise quantification, but the approximate lifetime risk of bipolar disorder in relatives of a bipolar proband are: monozygotic co-twin 40-70%; first degree relative 5-10%; unrelated person 0.5 1.5%. Occasional families may exist in which a single gene plays the major role in determining susceptibility, but the majority of bipolar disorder involves the interaction of multiple genes (epistasis) or more complex genetic mechanisms (such as dynamic mutation or imprinting). Molecular genetic positional and candidate gene approaches are being used for the genetic dissection of bipolar disorder. No gene has yet been identified but promising findings are emerging. Regions of interest identified in linkage studies include 4p16, 12q23-q24, 16p13, 21q22, and Xq24-q26. Chromosome 18 is also of interest but the findings are confusing with up to three possible regions implicated. To date most candidate gene studies have focused on neurotransmitter systems influenced by medication used in clinical management of the disorder but no robust positive findings have yet emerged. It is, however, almost certain that over the next few years bipolar susceptibility genes will be identified. This will have a major impact on our understanding of disease pathophysiology and will provide important opportunities to investigate the interaction between genetic and environmental factors involved in pathogenesis. This is likely to lead to major improvements in treatment and patient care but will also raise important ethical issues that will need to be addressed. PMID- 10465109 TI - Hydrometrocolpos and polydactyly: a common neonatal presentation of Bardet-Biedl and McKusick-Kaufman syndromes. AB - McKusick-Kaufman syndrome (MKKS) is a rare, recessively inherited syndrome reported mainly in young children and is characterised by vaginal atresia with hydrometrocolpos, postaxial polydactyly, and congenital heart defect. Bardet Biedl syndrome (BBS) is the generic name for a genetically heterogeneous group of autosomal recessive disorders characterised by retinal dystrophy or retinitis pigmentosa (appearing usually between 10 and 20 years of age), postaxial polydactyly, obesity, nephropathy, and mental disturbances, or, occasionally, mental retardation. Typically, MKKS is diagnosed (and reported) in very young children, whereas the diagnosis of BBS often is delayed to the teenage years. We report here a series of nine patients diagnosed in infancy with MKKS because of the presence of vaginal atresia and postaxial polydactyly, who later developed obesity and retinal dystrophy, thus turning out to be instances of BBS. The overlap of BBS and MKKS is a real diagnostic pitfall and its importance has to be stressed, for genetic counselling, for clinical management and follow up, and for molecular approaches. The diagnosis of MKKS should be considered with caution in all published cases described exclusively in the neonatal period and in those with mental retardation. We strongly recommend all children seen in infancy with a diagnosis of MKKS to be re-evaluated for RP and other signs of BBS. PMID- 10465110 TI - Familial cryptic translocation between chromosomes 2qter and 8qter: further delineation of the Albright hereditary osteodystrophy-like phenotype. AB - Recently five patients with an Albright hereditary osteodystrophy (AHO)-like phenotype were reported to have a subtelomeric deletion of the long arm of chromosome 2. These patients showed a striking resemblance to a number of patients from a large pedigree known to us for a long time. After molecular confirmation of a subtelomeric deletion in one patient, FISH analysis was used and a cryptic translocation between the long arms of chromosomes 2 and 8, t(2;8)(q37.3;q24.3), was detected. Remarkably, five proven and 10 probable cases with a 2qter deletion were found in the family, but none with an 8qter deletion. This was not explained by increased fetal loss. The major clinical characteristics of terminal 2q deletion are a short, stocky build, round face, sparse hair, deeply set eyes, bulbous nose, thin vermilion border, brachymetaphalangism, seizures, and developmental delay. A specific behavioural phenotype consisting of periods of hyperkinesia and aggression can develop with age. The overall phenotype is sufficiently characteristic to allow clinical recognition. The cytogenetic and molecular studies did not narrow down the common deleted region. Both testing of additional 2q markers and characterisation of other AHO-like patients with 2q37 microdeletions may help to define the candidate gene region. PMID- 10465112 TI - Carrier testing of children for two X linked diseases in a family based setting: a retrospective long term psychosocial evaluation. AB - The question of whether genetic carrier testing should be performed on children has been the subject of much debate. However, one important element has been lacking from this debate. There has been practically no knowledge of how those tested in childhood have experienced carrier testing. Twenty three subjects in families affected by Duchenne muscular dystrophy and 23 in families affected by haemophilia A, all of whom had been tested during childhood for carriership in the Department of Medical Genetics, University of Helsinki, from 1984 to 1988, participated in our study. We investigated long term psychosocial consequences of carrier testing in childhood. A questionnaire relating to sociodemographic background and life situation was used, together with assessment of health related quality of life (HRQOL) using the RAND 36 item Health Survey 1.0 (RAND). RAND results showed that the emotional, social, and physical well being of the young female subjects was not statistically different from those of control female subjects at a similar age. We also found no statistically significant differences in means in any RAND dimension (p<0.146) between carriers, non carriers, and a group in which carrier status was uncertain. However, two out of seven carriers reported that they were worried and three that they were slightly worried about the test result. Four out of 22 young female subjects in the uncertain group reported being worried and 11 reported being slightly worried. PMID- 10465111 TI - Mutational analysis of the N-ras, p53, p16INK4a, CDK4, and MC1R genes in human congenital melanocytic naevi. AB - Eighteen human congenital melanocytic naevi (CMN) from 17 patients were screened for activating point mutations in the oncogenes N-ras and CDK4 and for sequence variants in the MC1R gene by combined RFLP-PCR/SSCP analysis. In addition, all lesions were screened for deletions and point mutations in the tumour suppressor genes p53 and p16INK4a (CDKN2A) by combined multiplex PCR/SSCP analysis. Positive screening data were specified by sequencing of the corresponding PCR product. Activating point mutations in the N-ras gene (nine CAA (Gln) to AAA (Lys) transversions and one CAA (Gln) to CGA (Arg) transition at codon 61) were detected at high frequency (56%). Furthermore, three missense mutations (V92M) and two silent mutations (CGA (Arg) to CGG (Arg), codon 213, exon 6) were found in the MC1R and p53 genes, respectively. No mutations were found in p16 or CDK4. The activated N-ras oncogene, which is also found in human cutaneous melanomas, may constitute a potential risk factor for melanoma formation within CMN. PMID- 10465113 TI - Recessively inherited multiple epiphyseal dysplasia with normal stature, club foot, and double layered patella caused by a DTDST mutation. AB - We have observed over 25 different mutations in the diastrophic dysplasia sulphate transporter gene (DTDST) in association with the recessive disorders achondrogenesis 1B, atelosteogenesis 2, and diastrophic dysplasia. The c862t (R279W) transition is the most common mutation in non-Finnish patients, but in these disorders it is usually combined with other DTDST mutations. We had not seen a case of homozygosity for c862t (R279W) until we analysed DNA from a 36 year old male with tall-normal stature (180 cm) who asked for genetic counselling for suspected multiple epiphyseal dysplasia. He was treated for club foot and hip dysplasia at birth. Skeletal changes consistent with multiple epiphyseal dysplasia, with the peculiar finding of a double layered patella, were recognised during childhood. Cleft palate, swelling of the ear pinna, and hitch hiker thumb were absent. He was found to be homozygous, and both healthy parents heterozygous, for the R279W mutation in DTDST, and his fibroblasts showed a sulphate incorporation defect typical of DTDST disorders. Counselling was given for a recessive disorder, thereby considerably reducing the probability of affected offspring. Multiple epiphyseal dysplasia is more frequently caused by dominant mutations in the COMP (EDM1, McKusick 132400) and COL9A2 genes (EDM2, McKusick 600204). A few other patients and families with features similar to our proband have been described previously and considered to have autosomal recessive MED (EDM4, McKusick 226900). This observation confirms the existence of this entity and assigns it to the phenotypic spectrum associated with mutations at the DTDST locus. PMID- 10465114 TI - Simultaneous occurrence of two supernumerary autosomal ring chromosomes r(1) and r(16) in twins. AB - Ring chromosomes are estimated to occur in 3/10000 newborns and the simultaneous occurrence of two autosomal rings must be a very rare event. Recently, the characterisation of these markers using fluorescence in situ hybridisation (FISH) has greatly enhanced cytogenetic-phenotypic correlations in patients with these marker chromosomes. This kind of analysis enabled us to clarify a unique karyotype containing a r(1) and a r(16) in identical twins born after a 26 week gestation with minimal somatic abnormalities. The origin of the rings was identified using a satellite and whole chromosome painting probes. FISH analysis showed the same abnormal female karyotype in both twins, 48,XX,+r(1)(p13q21),+r(16)(p11q11).ish r(1) (D1Z5+,wcpl+), r(16)(D16Z2+,wcp16+) in about two thirds of the cells. Each also had minor clones with a normal female karyotype or with one or the other supernumerary ring. Half of the r(1) contained CBG band negative material and the r(16) appeared to be totally CBG band positive. These twins represent the second report of the simultaneous occurrence of multiple autosomal rings. Their description may help to delineate a new chromosome disorder and shows the usefulness of FISH analysis. PMID- 10465115 TI - Two pairs of proven monozygotic twins discordant for familial amyloid neuropathy (FAP) TTR Met 30. AB - Twin studies are an important tool in medical genetics for the evaluation of the relative roles of genetic and non-genetic factors in several diseases. Familial amyloidotic polyneuropathy type I (FAP-I), TTR Met 30, was present in two sets of proven monozygotic (MZ) twins, one from Majorca and the other from Portugal. Monozygosity was established by analysis of DNA polymorphisms. Both pairs were discordant for age at onset and some clinical manifestations of FAP-I. We reviewed the differences in age at onset and clinical features in both sets and in two other pairs of presumed MZ twins with FAP-I and compared them with those in MZ twin pairs with other Mendelian disorders, such as neurofibromatosis type 1, Huntington's disease, facioscapulohumeral muscular dystrophy, and myotonic dystrophy. We conclude that, in addition to the postulated modifying genes, there must be a significant contribution from non-genetic factors to the phenotypic variability of FAP-I (age at onset and clinical expression), either because of environmental differences or stochastic events during (or after) the twinning process. PMID- 10465116 TI - Maternal uniparental disomy of chromosome 14 confined to an interstitial segment (14q23-14q24.2). AB - Maternal uniparental disomy for the complete long arm of chromosome 14 has been reported in 14 patients to date and is associated with a specific pattern of malformation. We report a child with clinical features of this syndrome who exhibits maternal uniparental disomy confined to a specific interstitial segment of chromosome 14. PMID- 10465117 TI - Microcephaly, microphthalmia, congenital cataract, optic atrophy, short stature, hypotonia, severe psychomotor retardation, and cerebral malformations: a second family with micro syndrome or a new syndrome? AB - We report on four children of both sexes from a highly inbred family with hypotonia, spastic diplegia, microcephaly, microphthalmia, congenital cataract, optic atrophy, ptosis, kyphoscoliosis, short stature, severe mental retardation, and cerebral malformations. Six other children may also have been affected. The differential diagnosis and the possibility of a second family with the micro syndrome are discussed. PMID- 10465118 TI - A syndrome of immune complex glomerulonephritis and ophthalmic abnormalities. AB - Two sibs (one male and one female) suffering from a combination of immune complex glomerulonephritis and various ophthalmologic disorders are presented. The two cases belong to a family in which the parents are not related and seven sibs are affected, three females and a male with the combination, and three males with severe ophthalmological changes and proteinuria. Clinically, case 2 had only ophthalmological manifestations but renal biopsy findings were similar to those of case 1, which could mean that all the others with eye abnormalities also had renal disease. Although there are several reports of combinations of eye and renal disorders, the sibs reported here do not fit into any of the known syndromes. PMID- 10465119 TI - Sequence analysis of the homogentisate 1,2 dioxygenase gene in a family affected by alkaptonuria. PMID- 10465120 TI - A new family linked to the RP1 dominant retinitis pigmentosa locus on chromosome 8q. PMID- 10465121 TI - 21-Hydroxylase deficiency in Italy: a distinct distribution pattern of CYP21 mutations in a sample from southern Italy. PMID- 10465122 TI - Identification of a frameshift mutation in the gene TWIST in a family affected with Robinow-Sorauf syndrome. PMID- 10465123 TI - Unexpected Angelman syndrome molecular defect in a girl displaying clinical features of Prader-Willi syndrome. PMID- 10465124 TI - Effects of order of magnesium exposure on the postantibiotic effect and bactericidal activity of ciprofloxacin. AB - Quinolone antibiotics are known to form chelates with various metal cations. It has also been recognized that the physicochemical properties of the chelated antibiotic differ significantly from its unchelated form, thereby causing a reduction in antimicrobial activity. In addition, the formation of metal chelates is also believed to be the reason for the significant reduction in oral bioavailability for the quinolones when concomitantly dosed with oral cation containing agents. This has prompted the adoption of an alternate dosing regimen by introducing an adequate interval between the two. As a result of this dosing strategy, pathogens are exposed to the quinolones and metal cations in alternate orders. Using magnesium, ciprofloxacin, and Escherichia coli as the test organism, investigations were conducted to study the changes in bactericidal activity and postantibiotic effect (PAE) in relation to the orders of cation/antibiotic exposure. Results showed a parallel decrease in both bactericidal activity and PAE when the test organism was exposed to the two agents simultaneously; however, no apparent influences on these two antimicrobial effects were observed when Mg2+ was presented before or after ciprofloxacin exposure. In line with the current dosing recommendations, the interval spaced between ciprofloxacin and Mg2+ should preserve both the bactericidal activity and PAE exhibited by the antibiotic. How the present data are to be extrapolated to other quinolones and cations should be the subject of future studies. PMID- 10465125 TI - Endotoxin release due to ciprofloxacin measured by three different methods. AB - Antibiotics are known to induce the release of bioactive endotoxin (LPS) from gram-negative bacterial cells. Because varying data have been published on the influence of quinolone antibiotics on LPS liberation, we studied the effect of ciprofloxacin on a culture of Escherichia coli by determining bacterial killing and free LPS concentrations in comparison with imipenem and ceftazidime. LPS levels were measured by three different methods, namely (1) the Limulus amebocyte lysate test, (2) an ELISA method based on capture of LPS by monoclonal antibodies, and (3) indirect determination by measuring the ability of antibiotic induced LPS to trigger TNFalpha release from a monocytic cell line. With both the Limulus and ELISA tests, a low endotoxin-releasing activity of ciprofloxacin was confirmed. In contrast to previous studies, this LPS also had low bioactivity in terms of TNFalpha induction. Limulus LPS determinations correlated more precisely with LPS bioactivity than did ELISA values, an observation which underlines the crucial role of LPS determination methods in studies of antibiotic-induced LPS release. PMID- 10465126 TI - Antimicrobial survey of urinary tract isolates from a pediatric department. AB - The epidemiology of urinary tract colonization/infection in children admitted during 1996 to the Pediatric Department of the University of Verona has been studied; 501/1959 urine cultures were positive (25.57%). 584 microorganisms (64.89% Gram-negative, 24.82% Gram-positive, 10.27% fungi) were isolated. The highest rate of Gram-negative isolation (80.0%) was observed in infants, while the highest rate of Gram-positive isolation (29.6%) was found in newborns admitted to the neonatal intensive care unit. Escherichia coli was the most frequently isolated microorganism in infants and children, but not in newborns. A 3-fold increase in resistant E. coli strains to cotrimoxazole/sulfamethoxazole and amoxicillin/clavulanic acid was documented during the last 3 years in this pediatric population. Our observations underline the importance of the survey of microbial maps in pediatric departments in order to optimize therapeutic and preventive choices. PMID- 10465127 TI - Development of bacterial resistance to the third generation cephalosporins and their clinical use. AB - Development of Gram-negative rods resistance to the third generation cephalosporins (cefotaxime, ceftazidime, cefoperazone) in connection with their application at the University Hospital in Olomouc was evaluated in this study. The highest increase in resistance to cefotaxime was detected in Enterobacter cloacae (from 22.9% in 1995 to 49.0% in 1997) and Enterobacter agglomerans strains (28.0% - 40.5%). In addition, increased resistance to ceftazidime in Acinetobacter baumannii (12.5% - 35.1%), Enterobacter aerogenes (7.4% - 20.9%), Enterobacter cloacae (16.7% - 47.2%) and Pseudomonas aeruginosa (4.0% - 26.3%) was observed. Finally, the greatest increase in frequency of strains resistant to cefoperazone was observed in E. aerogenes (18.4% - 30.1%), E. agglomerans (31.0% 52.3%), E. cloacae (35.5% - 47.2%) and Providencia rettgeri (26.5% - 53.2%). A 23.5% increase in third generation cephalosporin use was evident by evaluation of RDDD(ATB) parameters in 1996 and 1997. Corresponding values for individual antibiotics were 26.5% cefotaxime, 20.7% ceftriaxone, and 40.3% ceftazidime increase. However, cefoperazone use decreased by 10.9%. PMID- 10465128 TI - Influence of comorbidity and severity on the clinical outcome of bacteremic pneumococcal pneumonia treated with beta-lactam monotherapy. AB - The influence of the severity of pneumonia and comorbidity factors, as predictors of clinical outcome, was assessed in patients with microbiologically documented pneumococcal bacteremic pneumonia treated with penicillin or third generation cephalosporin monotherapy in a 5-year retrospective study. Among 288 patients admitted to three Spanish hospitals with bacteremic pneumococcal pneumonia, 65 (23%) were included. Twenty-four were treated with penicillins and 41 with a third-generation cephalosporin. Twenty-seven patients (42%) had severe pneumonia and 41 (63%) had a comorbidity index >1. Twenty-one patients (32%) were infected with penicillin-resistant strains. Four cases (2 with penicillin-resistant strains; 3 treated with cephalosporins) were clinical failures. Four cases (3 with penicillin-resistant strains; 2 treated with cephalosporins) died, i.e. 6% mortality rate. The only factor that influenced empirical treatment election was HIV-positive condition. Clinical outcome was not influenced by treatment election, penicillin susceptibility of the infecting pneumococci, patient basal conditions or severity of pneumonia, but the latter was associated with mortality and length of hospitalization. PMID- 10465129 TI - A randomized trial of cefepime and ceftazidime for the treatment of community acquired pneumonia. AB - We compared the effectiveness and safety of ceftazidime and cefepime in hospitalized patients with community-acquired pneumonia. The 148 enrolled patients received 2 g ceftazidime three times daily or 2 g cefepime twice daily. The clinical success rate was the same for both drugs. Even the microbiological effectiveness was similar. Both drug regimens were well tolerated. We conclude that 2 g ceftazidime three times daily were as effective as 2 g cefepime twice daily for the treatment of community-acquired pneumonia in hospitalized patients. The cost of ceftazidime treatment was, however, higher than the cost of cefepime treatment. PMID- 10465130 TI - Cefepime as empirical monotherapy in febrile patients with hematological malignancies and neutropenia: a randomized, single-center phase II trial. AB - The purpose of this phase II trial was to evaluate the efficacy and safety of cefepime monotherapy in patients with neutropenia expected to last more than 7 days. Sixty-nine patients with neutropenia (<0.5 x 10(9)/1) were randomized during 94 episodes of fever to receive either cefepime monotherapy (n=76) or combination therapy with trimethoprim/sulfamethoxazole plus amikacin (TMP/SMZ plus AMI, n=18). A successful response to cefepime was seen in 31/76 (41%) episodes, with 10/36 (28%) in microbiologically documented infections, 3/4 (75%) in clinically documented infections and 18/36 (50%) in fever of unknown origin. No patient in either treatment group died due to the presenting infection. One patient in the cefepime group discontinued treatment due to a rash. Susceptibility testing of blood isolates by E-test strip showed low MIC values to cefepime for most isolates. It is concluded that cefepime monotherapy appeared both safe and effective as empirical therapy in patients with febrile neutropenia. PMID- 10465131 TI - An open evaluation of triple antibiotic therapy including vancomycin for febrile bone marrow transplant recipients with severe neutropenia. AB - Infectious complications still represent a major problem in patients submitted to bone marrow transplant (BMT); approximately 40% of febrile episodes are associated with infection and one-third of these are bacteremias. Opinions about the best appropriate empiric regimens are based on evaluation of cost, potential for adverse side-effects, development of bacterial resistance, prevalent nosocomial infections. In order to assess the clinical and microbiological effectiveness of an aggressive approach, we performed a prospective open study in 72 neutropenic febrile BMT patients, employing a triple antibiotic association including amikacin 500 mg x 8h, ceftazidime 2 g x 8 h, vancomycin 500 mg x 8 h as first-line empiric treatment. For the purpose of this study, a lasting return of temperature to normal and complete disappearance of either clinical or bacteriological signs of infection without any modification of therapy was considered as success; the persistence of fever after 72 hours or a protocol change was considered as failure. Eighty episodes were enrolled during the course of the study; bacteriological evidence of infection was obtained in 23 (28.7%) febrile episodes. Median duration of antibiotic administration and of febrile episodes were 5 and 2 days respectively. Overall response rate based on clinical responses was 87% and 91% in microbiological documented infections. Death due to sepsis nor toxicity were observed. This triple antibiotic combination appears to be a very effective regimen for the empiric treatment of febrile episodes in severely neutropenic BMT recipients. PMID- 10465132 TI - Cd 34+ cells and clonogenicity of peripheral blood stem cells during chemotherapy treatment in association with granulocyte colony stimulating factor in osteosarcoma. AB - The introduction of aggressive chemotherapy in the treatment of osteosarcoma has improved the long-term outcome for these patients. With the increasing aggressiveness of chemotherapy protocols, hematopoietic growth factors have emerged as useful adjuncts involving, in some cases, rescue by peripheral blood stem cell (PBSC) infusion to assist faster recovery and maintain relative dose intensity. To evaluate the number of PBSCs needed, we analyzed the number of CD34+ cells and hematopoietic progenitor cells in the peripheral blood of 16 patients with osteoblastic, condroblastic and fibroblastic osteosarcoma enrolled in an Istituto Ortopedico Rizzoli-Scandinavian Sarcoma Group (IOR-SSG) pilot study, consisting of two cycles of preoperative high dose chemotherapy. The blood samples were studied at different times. The CD34+ cells were analyzed by flow cytometry and the hematopoietic progenitor cells were analyzed by tissue culture clonogenic assay. In comparing the two courses of chemotherapy, we observed that modification of the mean values of WBC, CD34+ and CFU-GM were very similar. The second course of chemotherapy seemed to induce greater hematological toxicity. All three parameters showed good correlation. The results demonstrated that the best time to collect PBSC by means of leukapheresis is post G-CSF used as rescue after ifosfamide treatment. We verified the ability of G-CSF to mobilize PBSCs in patients with osteosarcoma through cytofluorimetric analysis of CD34+ cells and their clonogenic capability. Moreover, during this preoperative treatment, we identified the best time to collect a sufficient number of PBSCs, that is after 9 10 days of G-CSF treatment following the first cycle of ifosfamide. PMID- 10465133 TI - Paclitaxel efficacy and tolerability in second-line treatment of refractory and relapsed ovarian cancer patients. AB - Nineteen patients with recurrent or refractory ovarian carcinoma after a first line platinum-based chemotherapy were treated with a 3-hour i.v. infusion of paclitaxel 175 mg/m2 every 3 weeks from November 1992 to October 1996. The major hematologic toxicity was neutropenia (63.2%). No febrile neutropenia was observed. Other hematologic effects were leukopenia (47.4%) and anemia (47.4%). The main non-hematologic toxicities were as follows: neuropathy (52.6%), nausea and vomiting (36.8%), myalgia (36.8%), cardiac toxicity (15.8%) and mucositis (10.5%). Alopecia was observed in the majority of cases. The overall response rate was 47.4%, with 5 (26.3%) complete responses (CRs) and 4 (21.1%) partial responses (PRs). The median duration of response was 7 months (range: 3-19), with a median follow-up of 17 months (range: 3-61). Quality of life of responding patients was good. Our results confirm that paclitaxel as second-line therapy in relapsed and refractory ovarian cancer patients is an acceptable treatment with a good safety profile, and can be safely administered at the dose of 175 mg/m2. In our study paclitaxel was more active in relapsed than in refractory patients. Consequently, further studies are needed to identify more effective drugs for the refractory subset. PMID- 10465134 TI - A phase II study of a three-drug combination (cisplatin, ifosfamide and vinorelbine) plus granulocyte-colony stimulating factor in advanced non small cell lung cancer. AB - Twenty-nine patients with advanced non-small-cell lung cancer (NSCLC) were treated with a combination of cisplatin 20 mg/m2 days 1-3, ifosfamide 1500 mg/m2 days 1-2 (plus mesna as uroprotector) and vinorelbine 25 mg/m2 days 1 and 5; filgrastim was given at the dose of 300 microg subcutaneously from day 8 to day 15. A response rate of 28% was observed. The activity of this combination in an outpatient setting, with acceptable toxicity, has been demonstrated. PMID- 10465135 TI - Management of anal cancer: our experience. AB - The aim of this report was to relate our experience in the management of anal cancer. Up to now, the standard treatment reported in the literature for this particular disease is combination chemo-radiotherapy in most patients. Management decisions such as radical chemotherapy, surgical resection for poor response or relapses are frequently modified by some factors such as disease stage, social and psychological status of the patient, age-associated comorbidity factors. From 1992 to 1998, 24 consecutive patients (median age: 64 years) with untreated epidermoid carcinoma of the anus (T2, 58%; T3, 42%; N+, 25%) were treated, as first-line therapy, with a simultaneous chemo-radiotherapy combination. The crude mortality rate was 34%, with 29% dying of their disease. The stage distribution and the amount of chemo-radiotherapy administered were not age-limited but younger patients had more surgery and suffered less toxicity with a greater proportion remaining alive and disease-free. These data suggest that a more aggressive multi-modality approach may improve disease-free response and survival. PMID- 10465136 TI - Clinical remission of xeroderma pigmentosum-associated squamous cell carcinoma with isotretinoin and chemotherapy: case report. AB - We report the case of a 7-year old boy with xeroderma pigmentosum and a large squamous cell carcinoma of the cheek. He received a combination of isotretinoin (1 mg/kg/day) and chemotherapy for a period of 3 months and showed complete remission of the tumor. Treatment modalities of malignancies in xeroderma pigmentosum are reviewed and discussed in relation to the literature. The advantages of our protocol were emphasized because of the rapid improvement in a short time with minimal side effects. PMID- 10465137 TI - Use and monitoring of bromides in epilepsy treatment. AB - Bromides are no longer a mainstay of epilepsy therapy because of the significant toxicity associated with their use and the availability of safer agents. However, bromides occasionally find a niche in the treatment of patients with refractory seizures, particularly in pediatrics. When the decision to utilize this therapy is made, the clinician may be frustrated by the lack of concise, current information regarding bromides. This review provides an update on the mechanism of action, efficacy, pharmacokinetics, dose, and adverse effects of bromides. PMID- 10465138 TI - Epilepsy syndromes in patients with childhood-onset seizures in Finland. AB - Childhood-onset epilepsy is a common disorder. There is insufficient data on the distribution of epilepsy syndromes in the population and their effect on long term prognosis. This report presents the data on epilepsy syndromes in a childhood-onset epilepsy cohort. A population-based active-prevalence cohort of all children under 16 years of age residing in the catchment area of Turku University Hospital with active epilepsy between 1961 and 1964 was monitored prospectively until 1992. Etiology, seizure type, and epilepsy syndromes were classified according to the recent guidelines of the International League Against Epilepsy. The etiology of seizures was idiopathic in 28%, cryptogenic in 22%, and remote symptomatic in 50%. Seizures were classifiable in 235 patients (96%) and included 157 (64%) with partial seizures and 88 (36%) with generalized seizures. Epilepsy syndromes could be classified in 223 patients (91%) and included 152 (62%) localization-related syndromes, 61 (25%) generalized epilepsy syndromes, and 10 (4%) with syndromes not determined as being focal or generalized. Prognosis for both survival and remission was dependent on etiology and on the specific epilepsy syndrome. The authors conclude that the International League Against Epilepsy guidelines can be successfully applied to a population-based cohort with childhood-onset epilepsy. Accurate classification of syndromes is important because in many cases long-term outcome may be largely determined by the underlying epilepsy syndrome. PMID- 10465139 TI - Skeletal muscle mitochondrial defects in nonspecific neurologic disorders. AB - A group of 25 children (5 months to 20 years of age) presenting with intractable seizures, developmental delay, and severe hypotonia, who did not fall into the known categories of mitochondrial encephalomyopathies, underwent muscle biopsy for evaluation of mitochondrial function and were compared with age-matched control subjects. Biopsied skeletal muscle was analyzed for six mitochondrial enzyme-specific activities, mitochondrial DNA point mutations and deletions, and mitochondrial DNA levels. The data reveal a high incidence of specific mitochondrial enzyme activity defects. Reduced activity levels were evident in complex I (11 patients), III (24 patients), IV (nine patients), and V (10 patients). Two patients also exhibited pronounced reduction in mitochondrial DNA levels (80% reduction compared with control subjects). Two patients manifested increased levels of 5-kb and 7.4-kb mitochondrial DNA deletions. Pathogenic mutations previously described in association with mitochondrial encephalomyopathies were not evident. The data suggest that mitochondrial dysfunction, including extensive defects in specific enzyme activities, may be frequently present in children with seizures, developmental delay, and hypotonia that do not fall within the known mitochondrial encephalomyopathies. These mitochondrial deficiencies can be primarily ascertained by biochemical analysis and are rarely accompanied by mitochondrial ultrastructural changes. The molecular basis of these defects, their role in these disorders, and potential treatment warrant further study. PMID- 10465140 TI - Lactate attenuates neuron specific enolase elevation in newborn rats. AB - This study was undertaken to investigate the protective role of lactate on the hypoxic brain in newborn rats. A total of 107 7-day-old Wistar rats were divided into three groups. The lactate accumulation group was given 5% oxygen and 95% nitrogen for 30 minutes. The lactate elimination group was given 5% oxygen, a concentration of 7.5% carbon dioxide, and 87.5% nitrogen for 30 minutes. The control rats were placed in room air. Lactate levels in the brain tissue were higher in the lactate accumulation group than in those of the control group (control: 1.78 +/- 0.91, lactate accumulation: 11.42 +/- 1.64 mmol/kg) and significantly decreased in the lactate elimination group (4.10 +/- 1.73 mmol/kg). Blood pH remained at the same levels in the two groups. Neuron specific enolase in the cerebrospinal fluid, which is the initial neurocyte damage marker, was significantly elevated in the lactate elimination group (control: 18.3 +/- 7.5, lactate accumulation: 18.8 +/- 7.9, lactate elimination: 63.1 +/- 61.3 ng/mL). Brain adenosine 5'-triphosphate levels were significantly decreased in the lactate elimination group. Histologic findings of the brain at 72 hours after the load revealed no abnormal changes in any of the groups examined. The authors conclude that lactate accumulation plays a protective role on the hypoxic brain in newborn rats. PMID- 10465141 TI - Ketogenic diet in the treatment of refractory epilepsy in childhood. AB - There has been renewed interest in the ketogenic diet in the treatment of medically refractory seizure disorders in childhood. This article reports the results of a retrospective chart review of 52 patients who were treated with the ketogenic diet. The vast majority (49 of 52) were treated with the classic 4:1 diet. Seizure control improved in 67.3% of patients with complete abolition of seizures in six. Adverse reactions were uncommon and included the development of renal stones, gall bladder stones, and hypoproteinemia in one patient each. Routine biochemical screening during the diet did not identify or prevent adverse events. The authors' experiences with the diet emphasize the need for close ongoing medical and dietary supervision. PMID- 10465142 TI - Pathogenetic role of monoamine metabolism in complex febrile seizures. AB - To investigate the pathogenetic role of monoamine metabolism for febrile seizures, concentrations of homovanillic acid and 5-hydroxyindoleacetic acid were evaluated in the lumbar cerebrospinal fluid of affected children. This series included children who underwent lumbar puncture within 4 hours after seizures, at which time the lumbar cerebrospinal fluid is presumed to represent a ventricular source of cerebrospinal fluid, without the consequence of a seizure. The subjects were 21 patients with simple febrile seizures, eight with complex febrile seizures, and eight control patients. The complex febrile seizure group had lower homovanillic acid and 5-hydroxyindoleacetic acid levels than the control group, whereas the simple febrile seizure group and control group did not have a significant difference. Although statistically not significant, the complex febrile seizure group was inclined toward lower homovanillic acid and 5 hydroxyindoleacetic acid levels than the simple febrile seizure group. These results implied that abnormal monoamine metabolism contributes to seizure susceptibility in the complex febrile seizure group. The simple febrile seizure group may be heterogeneous, and abnormal monoamine metabolism can play a role in seizure development for some children of this group. PMID- 10465143 TI - Neurologic outcomes of infants with tremor within the first year of life. AB - The neurologic outcomes of 24 infants with no other overt neurologic signs except for tremor that lasted through or began after the neonatal period were evaluated during a follow-up period of 3-10.2 years (mean = 6.8 years). Twenty patients had normal outcomes, three had borderline intelligence, and the remaining patient had cerebral palsy (to a minimum degree). The duration, incidence, persistence, and type of tremor were not related to the neurologic outcomes in this group. The outcomes of the 10 children with no perinatal complications were good. In contrast, among the 14 children with perinatal complications, four had mild neurologic deficits, as previously mentioned. The neurologic outcomes of infants with no other neurologic signs except for tremor seem to be generally good, especially when the infants have no perinatal complications. It appears that tremor in neonates and infants is a functional sign that can be caused by a variety of factors, and the neurologic outcomes are related directly to the underlying brain damage induced by perinatal insults. PMID- 10465144 TI - Neuroimaging findings of four patients with Sandhoff disease. AB - Sandhoff disease is a severe form of GM2 gangliosidosis that is caused by the deficiency of both hexosaminidase A and B. Startle reaction, hypotonia, psychomotor retardation, and blindness are the main clinical features. Presented are computed tomography and magnetic resonance imaging findings of four patients with Sandhoff disease diagnosed by enzymatic analyses. Bilateral homogeneous thalamic hyperdensity was evident on computed tomography. Magnetic resonance imaging scans revealed mild cortical atrophy, a thin corpus callosum, and abnormal signal intensities in the caudate nucleus, globus pallidum, putamen, cerebellum, and brainstem. No correlation was evident between the severity of the central nervous system imaging findings and the clinical pictures. In this article the neuroimaging findings of four patients with Sandhoff disease are discussed. PMID- 10465145 TI - Epilepsy and polymicrogyria in Kabuki make-up (Niikawa-Kuroki) syndrome. AB - Kabuki make-up syndrome is a rare dysmorphogenic disorder characterized by peculiar facial appearance (resembling the make-up of actors in Kabuki, the traditional Japanese theatre), skeletal anomalies, dermatoglyphic abnormalities, postnatal growth deficiency, and mental retardation. Central nervous system dysfunctions, other than mental retardation, are rarely reported; they include microcephaly, brachycephaly, early hypotonia, feeding disorders, subatrophy of the optic nerves, subarachnoid cyst, cerebellar and brainstem atrophy, and epilepsy. These manifestations appear to be more common in non-Japanese patients. Reported is an Italian child with phenotypical appearance of Kabuki make-up syndrome and partial epilepsy who demonstrated polymicrogyria on neuroimaging. This article is the first report of a gyration disorder in Kabuki make-up syndrome. The relationship between epilepsy and polymicrogyria in this patient is discussed. PMID- 10465146 TI - A left atrial myxoma presenting as a cerebrovascular accident. AB - A 14-year-old female presented with common clinical findings for a rare primary intracardiac tumor. Primary cardiac tumors are rare in all age groups, occurring in 0.05% of routine postmortem examinations. Pediatric primary cardiac tumors are likewise uncommon, with the most common being a rhabdomyoma. Atrial myxomas occur infrequently in the pediatric age group. They occur primarily between the third and sixth decade, making them the most common adult primary cardiac tumor. The following case presentation demonstrates a common clinical presentation for an intracardiac mass rarely diagnosed in the pediatric population. This patient's acute neurologic symptoms required prompt recognition of an intracardiac etiology. This recognition proved critical for the acute and long-term medical and surgical management of this patient. PMID- 10465147 TI - Sagittal sinus thrombosis associated with thrombocytopenia: a report of two patients. AB - Reported are two patients presenting with both thrombocytopenia and sagittal sinus thrombosis. The first patient is a 42-month-old male with no identified thrombophilic risk factors who developed acute neurologic symptoms after an acute infection. The second patient is a 22-month-old female with no history of preceding infection but had a positive lupus anticoagulant test. She also developed deep venous thrombosis and was treated with intravenous heparin. Both patients are currently doing well without neurologic deficits. To the authors' knowledge the second patient is the youngest reported patient with cerebral vein thrombosis associated with thrombocytopenia and lupus anticoagulant. These observations call attention to the need for a thorough investigation of thrombophilic risk factors in pediatric patients with thrombotic complications. PMID- 10465148 TI - Anterior lumbosacral radiculopathy after intrathecal methotrexate treatment. AB - Intrathecal chemotherapy with methotrexate or cytosine arabinoside is the standard approach to prophylaxis and treatment of central nervous system leukemia in children. Progressive paraplegia, one of the devastating neurologic complications related to this mode of treatment, has been attributed to spinal cord toxicity. Reported are three children who developed progressive paraparesis after intrathecal methotrexate administration followed by complete or partial recovery. Gadolinium enhancement of anterior lumbosacral spinal nerve roots was demonstrated in all three patients, and an elevation of cerebrospinal fluid immunoglobulin G synthesis was evident in two patients with more severe symptoms. The clinical data suggest that anterior lumbosacral radiculopathy is also a type of neurologic complication associated with intrathecal methotrexate treatment. PMID- 10465149 TI - Nonconvulsive status epilepticus in a child with congenital bilateral perisylvian syndrome. AB - A 9-year-old male with congenital bilateral perisylvian syndrome is described. He had pseudobulbar palsy, mental retardation, and intractable epilepsy. Computed tomography and magnetic resonance images of the brain demonstrated bilateral perisylvian malformations and a diffuse pachygyric appearance. At 8 years of age, he had episodes of excessive drooling, fluctuating impairment of consciousness, unsteady sitting, and frequent head drop that lasted several days. The electroencephalogram demonstrated continuous diffuse slow spike and waves. These findings suggested atypical absence status epilepticus. Intravenous administration of diazepam resulted in transient improvement of clinical and electroencephalographic findings. Status epilepticus recurred within several minutes after diazepam administration. Although no patient has been reported to have a history of status epilepticus among those affected by this syndrome, it seems that atypical absence status can occur more frequently than expected, as seen in Lennox-Gastaut syndrome. After recognition and confirmation of nonconvulsive status epilepticus, immediate treatment must be attempted. PMID- 10465150 TI - Intravenous immunoglobulin therapy in acute disseminated encephalomyelitis. AB - Three children ranging in age from 2 to 5 years with acute disseminated encephalomyelitis (ADEM) were successfully treated with high-dose intravenous immunoglobulin (IVIG). Their symptoms were somnolence, fever, headache, vomiting, and resting tremor. In all of these patients, it was difficult to distinguish the condition from viral encephalitis before analyzing the myelin basic protein. ADEM was diagnosed because of increased levels of myelin basic protein in their cerebrospinal fluid and abnormal high-signal intensity on T2-weighted magnetic resonance imaging. All patients were given IVIG at a dose of 400 mg/kg/day for 5 consecutive days. The patients rapidly regained consciousness in 14 hours, 2 days, and 4 days and demonstrated a complete clinical improvement within 18 days, 10 days, and 7 days of the initiation of the treatment, respectively. IVIG may prove useful as an alternative treatment to corticosteroids for ADEM. PMID- 10465151 TI - Stroke in pediatric HIV infection. PMID- 10465152 TI - The plasma membrane calcium pump, its role and regulation: new complexities and possibilities. AB - Significant progress has been achieved in elucidating the role of the plasma membrane Ca2(+)-ATPase in cellular Ca2+ homeostasis and physiology since the enzyme was first purified and physiology since the enzyme was first purified and cloned a number of years ago. The simple notion that the PM Ca2(+)-ATPase controls resting levels of [Ca2+]CYT has been challenged by the complexity arising from the finding of four major isoforms and splice variants of the Ca2+ pump, and the finding that these are differentially localized in various organs and subcellular regions. Furthermore, the isoforms exhibit differential sensitivities to Ca2+, calmodulin, ATP, and kinase-mediated phosphorylation. The latter pathways of regulation can give rise to activation or inhibition of the Ca2+ pump activity, depending on the kinase and the particular Ca2+ pump isoform. Significant progress is being made in elucidating subtle and more profound roles of the PM Ca2(+)-ATPase in the control of cellular function. Further understanding of these roles awaits new studies in both transfected cells and intact organelles, a process that will be greatly aided by the development of new and selective Ca2+ pump inhibitors. PMID- 10465153 TI - Accuracy of repeated blood sampling in rats: a new technique applied in pharmacokinetic/pharmacodynamic studies of the interaction between warfarin and co-enzyme Q10. AB - In the past, combined pharmacokinetic/pharmacodynamic (PK/PD) modeling studies of oral anticoagulants in rats have been hampered by the technical problems of blood sampling. In the present study, a semi-micromethod of preparing serial plasma samples for accurate assessment of the prothrombin times (PT) and clotting factor VII activity (CFA) in rats is proposed. The method consists of orbital bleeding, blood sample weighing, gravity calculation and buffer volume adjustment. No significant differences of CFA (percentage normal) were found between citrate diluted and undiluted plasma. This technique was employed to examine the possibility of PK/PD interaction between warfarin and Co-enzyme Q10 (CoQ10). Rats were given a single oral dose (1.5 mg/kg) of warfarin either alone or on day 4 of an 8-day oral dosing regimen of 10 mg/kg CoQ10 daily. Serial plasma and serum samples, which were subjected to respectively measurements of the anticoagulant effects and concentrations of warfarin and its main metabolites, were obtained over a 96-h period following warfarin administration. All rats survived the whole experiment and maintained a stable condition except for a marked hematocrit decrease. CoQ10 significantly augmented warfarin metabolism but showed little effect on the absorption of warfarin. CoQ10 alone had no apparent effect on either the PT or CFA. The concomitant administration of CoQ10 and warfarin does not significantly affect the anticoagulant effect of warfarin. In conclusion, the proposed serial orbital bleeding technique in rats to prepare an accurate citrate diluted plasma for PT and CFA measurement is rapid and reliable. PMID- 10465154 TI - Quantitation of tremor in response to beta-adrenergic receptor stimulation in primates: relationship with hypokalemia. AB - The primary adverse effect of stimulation of beta2-adrenergic receptors is elicitation of tremor. Tremor measurements in response to beta2-adrenergic receptor stimulation were performed in a quantitative manner using a modified miniature semiconductor accelerometer in African green monkeys. The accelerometer was taped to the middle finger tip of anesthetized monkeys, and recordings of onset, duration and peak tremor responses were obtained. The selective beta2 adrenergic agonist, salbutamol (0.5 mg/kg i.v.), caused a marked increase in tremor which started within 5 min following injection and lasted for approximately 60 min. The finger tremor response was not visible, but was measurable by the accelerometer, and the increase in tremor was significantly greater from baseline within 10 min. Plasma K+ concentrations were markedly decreased within the first 15 min and remained at low steady-state concentrations during the 60-min recordings. The tremor response was abolished by the selective beta2-adrenergic receptor antagonist, ICI-118551 (0.2 mg/kg). ICI-118551 caused a significant reversal of the plasma K+ decrease but the K+ levels remained higher than control levels. These studies demonstrate that stimulation of beta2 adrenergic receptors causes tremor, most likely from entry of K+ into skeletal muscle and that there is a direct correlation between tremor and hypokalemic response. PMID- 10465155 TI - Intrinsic activity and EC50: the simplest tools for the evaluation of the dissociation constant of a partial agonist. AB - In functional pharmacology, the Furchgott's analysis and the Waud's analysis are the most widely used among the reliable different methods actually available, for the determination of the dissociation constant of partial agonists. However, they need the application of relatively complicated procedures of manipulation and interpolation of raw data. On the basis of empirical assumptions, this article proposes a new approach, which probably can be considered the simplest method to determine the dissociation constant of a partial agonist, because of the rapid experimental protocol and the easy calculation procedure. Computer-generated concentration-response curves (CRC) for hypothesised partial agonists were analysed by the widely known Waud's analysis and by this new approach. Furthermore, this new analysis was also used to evaluate experimental data from literature, relative to the dissociation constants of alpha-adrenoceptor partial agonists, recorded in rabbit and rat aortae and calculated by the Waud's method. The results obtained by the new approach, both for the computer-generated and for the experimentally studied partial agonists, showed a high level of accuracy, when compared with the classical Waud's analysis. PMID- 10465156 TI - Muscle contraction-induced steal model in the anesthetized cat. AB - The current study describes a model in the anesthetized cat which may be used to investigate pharmacological interventions for the treatment of vascular "steal" during muscle contraction. An artificial stenosis was placed around both the descending abdominal aorta and the left external iliac artery. Arterial blood flow was measured in the right and left external iliac and right femoral arteries. Contraction of the left hindquarters, induced by electrical stimulation of the left sciatic and femoral nerves, resulted in an increase of left iliac artery blood flow from 14.7+/-2.0 to 30.5+/-2.8 ml min-1 (p<0.001). A simultaneous reduction of blood flow measured at the level of the right femoral artery (6.7+/-0.1 to 4.5+/-0.7 ml min(-1), p<0.01) was observed in the noncontracting hindlimb (y = 7.76 - 0.10x, correlation coefficient = -0.865). The magnitude of each response was reproducible within the same animal. Administration of 8-phenyltheophylline, a nonselective antagonist of adenosine receptors, reduced the functional hyperemia response within the left external iliac artery by 32% and reduced the "steal" from the vascular bed perfused by the right femoral artery by 21%. PMID- 10465157 TI - Suitability of the old fowl rectal caecum preparation for investigating the selectivity of beta-adrenergic drugs. AB - We tested alpha- and beta-adrenergic drugs on isolated strips of fowl rectal caecum from 14- to 16-week-old Warren hens. Basal tone and spontaneous motility were dose-dependently reduced by isoprenaline and all the selective beta-agonists tested (except xamoterol) with the following order of potency: isoprenaline=fenoterol=procaterol=clenbuterol>dobutamine> SR58611A. The results indicate that this tissue preparation consists almost entirely of beta2 adrenoceptors. This preparation may, therefore, be considered a suitable assay for discriminating beta1- from beta2-agonists according to their selectivity. PMID- 10465158 TI - Correlation of endothelium-dependent and -independent vasodilatation with liver function tests during prolonged perfusion of the rat liver. AB - Twelve male Wistar rats were anaesthetized with pentobarbitone (3 mg 100g(-1) i.p.), the livers were excised and perfused in vitro through the hepatic artery and portal vein at constant flow rates of 0.32+/-0.01 (mean+/-S.E.) and 0.98+/ 0.03 ml min(-1) g liver(-1), respectively. The tone of the preparation was raised by methoxamine (7.5 x 10(-6) M). Responses to mid-range doses of acetylcholine ( 11 log mol) and sodium nitroprusside (-9 log mol) produced submaximal degrees of vasodilatation (-log mol ED50 = 12.18+/-0.08) and (-log mol ED50 = 9.95+/-0.23), respectively, which did not subside until 5.5 h of perfusion. These did not coincide with the increase in activities of lactic acid dehydrogenase (LDH) and aspartate serine transaminase (AST) activity at 2.5 h, which were indicative of hepatocellular mitochondrial and cytoplasmic damage, respectively. Vascular responses suggested that there was little deterioration in endothelial or smooth muscle function in the hepatic artery up to 5 h perfusion. This model can be reliably used to investigate endothelium-dependent and -independent vasodilators in vascular pharmacological studies of the rat liver although some minimal increases may occur in AST and LDH activity before hemodynamic changes appear at 5.5 h. PMID- 10465159 TI - Analysis of ritonavir in plasma/serum and tissues by high-performance liquid chromatography. AB - A method has been developed to quantify ritonavir concentrations in human plasma and in mouse serum, liver, and brain using high-performance liquid chromatography. Extraction recoveries for ritonavir and its internal standard averaged greater than 95%. Within-day variability, expressed as a coefficient of variation, averaged 6% over the concentration range 0.5 microg/mL to 15 microg/mL ritonavir, and between-day variability averaged 5.6% over 5 microg/mL to 15 microg/mL ritonavir. The method was applied to quantitation of ritonavir in mouse serum and tissue. Measured values deviated less than 5% from the actual values in mouse serum, liver, and brain samples containing 5 microg/mL ritonavir. The slopes of calibration curves for extracted calf serum, mouse serum, mouse liver and mouse brain standards were nearly identical to the calibration slope of standards which were not extracted. All curves were linear through zero, and r2 was no less than 0.998 for any form of calibration. In addition, there was no chromatographic interference from commonly prescribed medications. PMID- 10465160 TI - Estimation of agonist affinity, using the slope of the concentration-effect curve: comment on the method of Calderone and Martinotti. AB - Recently, Calderone and Martinotti (J Pharmacol Toxicol Meth 40:57-62, 1998) presented a simple equation for the estimation of agonist dissociation equilibrium constants from data obtained from receptor inactivation experiments using an irreversible, competitive antagonist. In the present paper, however, it is demonstrated that application of this equation will result in significant over and underestimation of agonist affinity in the case of flat and steep concentration-effect curves, respectively. Therefore, accurate estimation of agonist affinity using irreversible receptor inactivation requires detailed curve shape information from both control and antagonist-treated tissues. PMID- 10465161 TI - Risking the eclipse. PMID- 10465162 TI - Emerging role of corticosteroids in chronic obstructive pulmonary disease. PMID- 10465163 TI - Do vitamin E and fish oil protect against ischaemic heart disease? PMID- 10465164 TI - Infant feeding and HIV-1 transmission. PMID- 10465165 TI - Batten's disease: eight genes and still counting? PMID- 10465166 TI - Brugada syndrome. PMID- 10465167 TI - Caveat doctor. PMID- 10465168 TI - Dietary supplementation with n-3 polyunsaturated fatty acids and vitamin E after myocardial infarction: results of the GISSI-Prevenzione trial. Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto miocardico. AB - BACKGROUND: There is conflicting evidence on the benefits of foods rich in vitamin E (alpha-tocopherol), n-3 polyunsaturated fatty acids (PUFA), and their pharmacological substitutes. We investigated the effects of these substances as supplements in patients who had myocardial infarction. METHODS: From October, 1993, to September, 1995, 11,324 patients surviving recent (< or = 3 months) myocardial infarction were randomly assigned supplements of n-3 PUFA (1 g daily, n=2836), vitamin E (300 mg daily, n=2830), both (n=2830), or none (control, n=2828) for 3.5 years. The primary combined efficacy endpoint was death, non fatal myocardial infarction, and stroke. Intention-to-treat analyses were done according to a factorial design (two-way) and by treatment group (four-way). FINDINGS: Treatment with n-3 PUFA, but not vitamin E, significantly lowered the risk of the primary endpoint (relative-risk decrease 10% [95% CI 1-18] by two-way analysis, 15% [2-26] by four-way analysis). Benefit was attributable to a decrease in the risk of death (14% [3-24] two-way, 20% [6-33] four-way) and cardiovascular death (17% [3-29] two-way, 30% [13-44] four-way). The effect of the combined treatment was similar to that for n-3 PUFA for the primary endpoint (14% [1-26]) and for fatal events (20% [5-33]). INTERPRETATION: Dietary supplementation with n-3 PUFA led to a clinically important and statistically significant benefit. Vitamin E had no benefit. Its effects on fatal cardiovascular events require further exploration. PMID- 10465169 TI - Oral corticosteroids in patients admitted to hospital with exacerbations of chronic obstructive pulmonary disease: a prospective randomised controlled trial. AB - BACKGROUND: The role of oral corticosteroids in treating patients with exacerbations of chronic obstructive pulmonary disease (COPD) remains contentious. We assessed in a prospective, randomised, double-blind, placebo controlled trial the effects of oral corticosteroid therapy in patients with exacerbations of COPD requiring hospital admission. METHODS: We recruited patients with non-acidotic exacerbations of COPD who were randomly assigned oral prednisolone 30 mg once daily (n=29) or identical placebo (n=27) for 14 days, in addition to standard treatment with nebulised bronchodilators, antibiotics, and oxygen. We did spirometry and recorded symptom scores daily in inpatients. Time to discharge and withdrawals were noted in each group. We recalled patients at 6 weeks to repeat spirometry and collect data on subsequent exacerbations and treatment. Hospital stay was analysed by intention to treat and forced expiratory volume in 1 s (FEV1) according to protocol. FINDINGS: FEV1 after bronchodilation increased more rapidly and to a greater extent in the corticosteroid-treated group: percentage predicted FEV1 after bronchodilation rose from 25.7% (95% CI 21.0-30.4) to 32.2% (27.3-27.1) in the placebo group (p<0.0001) compared with 28.2% (23.5-32.9) to 41.5% (35.8-47.2) in the corticosteroid-treated group (p<0.0001). Up to day 5 of hospital stay, FEV1 after bronchodilation increased by 90 mL daily (50.8-129.2) and by 30 mL daily (10.4-49.6) in the placebo group (p=0.039). Hospital stays were shorter in the corticosteroid-treated group. Groups did not differ at 6-week follow-up. INTERPRETATION: These data provide evidence to support the current practice of prescribing low-dose oral corticosteroids to all patients with non-acidotic exacerbations of COPD requiring hospital admission. PMID- 10465170 TI - Neuroimmune appendicitis. AB - BACKGROUND: 15-25% of appendices removed from patients with suspected appendicitis appear normal on histological examination. The cause of pain in such patients is unknown. Since the content of neuropeptides seems to be altered in chronic inflammation, we investigated possible changes in peptidergic innervation for substance P (SP), vasoactive intestinal peptide (VIP), and growth-associated protein-43 (GAP-43). METHODS: Appendices classified as showing acute appendicitis, non-acute appendicitis (clinical signs of acute appendicitis, but histologically not inflamed), or normal were processed for SP, VIP, and GAP-43 immunocytochemistry. The density of SP immunostaining was assessed by digitised morphometry. FINDINGS: 31 appendix specimens were studied (16 acute, 15 non acute). 16 specimens were used as controls. Expression of GAP-43 was increased in the non-acute appendices. We observed larger amounts of SP-immunoreactive and VIP immunoreactive nerves in the mucosal layer of the appendix in patients with non acute appendicitis than in controls and patients with acute appendicitis (mean % area SP-immunoreactive 0.0496 [SD 0.0113] non-acute, 0.0221 [0.0049] acute, 0.0229 [0.0068] controls). In addition, a close spatial relation between SP immunoreactive and VIP-immunoreactive nerve fibres and lymphoid cells was detected in the outer zone of lymph follicles. INTERPRETATION: Neuroproliferation in the appendix, in association with an increase in neurotransmitters SP and VIP, may be involved in the pathophysiology of acute right abdominal pain in the absence of an acute inflammation of the appendix. Our data, together with increasing knowledge about the way in which the nervous system and immune cells interact, suggest that neuroimmune appendicitis is a distinct pathological entity. PMID- 10465171 TI - Will elderly patients stand aside for younger patients in the queue for cardiac services? AB - BACKGROUND: Fair management of queues for hospital-based services requires consideration of clinical need, but there is no information on public attitudes towards non-clinical factors such as age or work status as determinants of priority. METHODS: We asked elderly residents of Padova, Italy, whether, if they were awaiting cardiac surgery or an outpatient cardiology consultant, they would give up their place in line for a younger or self-employed individual. We also elicited responses from a convenience sample of younger health workers asked to imagine themselves as elderly persons facing the same choices. FINDINGS: The eligible response rate was 72% (443/616). About half deemed it right to give up their place in line for cardiac surgery to a 45-year-old (51%, 95% CI 46-56) or self-employed person (47%, 95% CI 42-51) Proportions were significantly higher for an outpatient consultation (68%, 95% CI 63-72). In multivariate analyses, married respondents, those closer to age 65, university graduates, and those who were formerly self-employed were significantly less likely to respond affirmatively. In significant contrast for all four scenarios (p<0.0001), the overwhelming majority of non-elderly respondents refused to give up their places in line. INTERPRETATION: The majority of elderly citizens were hypothetically willing to cede priority in accessing cardiac care to younger or self-employed persons, but this willingness was attenuated among the "young" elderly and more privileged respondents. Non-elderly respondents were much less self-sacrificing, suggesting that ageing baby-boomers may be more assertive about their continuing rights to health care. PMID- 10465172 TI - Influence of infant-feeding patterns on early mother-to-child transmission of HIV 1 in Durban, South Africa: a prospective cohort study. South African Vitamin A Study Group. AB - BACKGROUND: The observation that mother-to-child transmission of HIV-1 can occur through breastfeeding has resulted in policies that recommend avoidance of breastfeeding by HIV-1-infected women in the developed world and under specific circumstances in developing countries. We compared transmission rates in exclusively breastfed, mixed-fed, and formula-fed (never breastfed) infants to assess whether the pattern of breastfeeding is a critical determinant of early mother-to-child transmission of HIV-1. METHODS: We prospectively assessed infant feeding practices of 549 HIV-1-infected women who were part of a vitamin A intervention trial in Durban, South Africa. The proportions of HIV-1-infected infants at 3 months (estimated by use of Kaplan-Meier life tables) were compared in the three different feeding groups. HIV-1 infection was defined by a positive RNA-PCR test. FINDINGS: At 3 months, 18.8% (95% CI 12.6-24.9) of 156 never breastfed children were estimated to be HIV-1 infected compared with 21.3% (17.2 25.5) of 393 breastfed children (p=0.5). The estimated proportion (Kaplan-Meier) of infants HIV-1 infected by 3 months was significantly lower for those exclusively breastfed to 3 months than in those who received mixed feeding before 3 months (14.6% [7.7-21.4] vs 24.1% [19.0-29.2], p=0.03). After adjustment for potential confounders (maternal CD4-cell/CD8-cell ratio, syphilis screening test results, and preterm delivery), exclusive breastfeeding carried a significantly lower risk of HIV-1 transmission than mixed feeding (hazard ratio 0.52 [0.28 0.98]) and a similar risk to no breastfeeding (0.85 [0.51-1.42]). INTERPRETATIONS: Our findings have important implications for prevention of HIV-1 infection and infant-feeding policies in developing countries and further research is essential. In the meantime, breastfeeding policies for HIV-1-infected women require urgent review. If our findings are confirmed, exclusive breastfeeding may offer HIV-1-infected women in developing countries an affordable, culturally acceptable, and effective means of reducing mother-to child transmission of HIV-1 while maintaining the overwhelming benefits of breastfeeding. PMID- 10465173 TI - Effect of idebenone on cardiomyopathy in Friedreich's ataxia: a preliminary study. AB - BACKGROUND: Friedreich's ataxia is caused by a deficiency of frataxin, a protein involved in regulation of mitochondrial iron content. We have reported a combined deficiency of a Krebs-cycle enzyme, aconitase, and three mitochondrial respiratory-chain complexes in endomyocardial biopsy samples from patients with this disorder. All four enzymes share iron-sulphur cluster-containing proteins that are damaged by iron overload through generation of oxygen free radicals. We used an in-vitro system to elucidate the mechanism of iron-induced injury and to test the protective effects of various substances. On the basis of these results, we assessed the effect of idebenone (a free-radical scavenger) in three patients with Friedreich's ataxia. METHODS: Heart homogenates from patients with valvular stenosis were tested for respiratory-chain complex II activity, lipoperoxidation, and aconitase activity by spectrophotometric assays, in the presence of reduced iron (Fe2+), oxidised iron (Fe3+), desferrioxamine, ascorbic acid, and idebenone. The Friedreich's ataxia patients (aged 11 years, 19 years, and 21 years) underwent ultrasonographic heart measurements at baseline and after 4-9 months of idebenone (5 mg/kg daily). FINDINGS: Fe2+ (but not Fe3+) decreased complex II activity and increased lipoperoxidation in heart homogenate. Addition of ascorbate or desferrioxamine increased some of the iron-induced adverse effects. Idebenone protected against these effects. In the three patients, left ventricular mass index decreased from baseline to 4-9 months of idebenone treatment (patient 1, 145 g to 114 g; patient 2, 215 g to 151 g; patient 3, 408 g to 279 g). INTERPRETATION: Our in-vitro data suggest that both iron chelators and antioxidant drugs that may reduce iron are potentially harmful in patients with Friedreich's ataxia. Conversely, our preliminary findings in patients suggest that idebenone protects heart muscle from iron-induced injury. PMID- 10465174 TI - Diabetes but not psoriasis. PMID- 10465175 TI - Nephropathy caused by Chinese herbs in the UK. AB - The use of Chinese herbal remedies is increasing in the UK. We report the presence of a nephrotoxic compound in herb samples, which led to end-stage renal failure in two patients. We suggest that use of these products is regulated more tightly. PMID- 10465176 TI - Altered lung antioxidant status in patients with mild asthma. AB - Lung lining fluid ascorbic acid (vitamin C) and alpha-tocopherol (vitamin E) concentrations are low in patients with mild asthma even though blood levels are normal or increased. These findings, along with the presence of increased amounts of oxidised glutathione in their airways, indicate that patients with asthma are subject to increased oxidative stress. PMID- 10465177 TI - Should radiologists image and report sacroiliac joints in patients referred for examination of the lumbar spine? AB - It is routine to image the sacroiliac joints (SIJs) in patients referred for radiographic examination of the lumbar spine but there were only two consistent with ankylosing spondylitis in a review of 392 radiographic reports of lumbar spine examinations. A more sensible policy would be to only image and report on the SIJs if the clinical history were indicative of ankylosing spondylosis. PMID- 10465178 TI - Pesticide exposure and decreased fertilisation rates in vitro. AB - The effect of paternal occupational exposures on fertilising ability was investigated in 836 couples who sought in-vitro fertilisation treatment. Fertilisation rates were significantly decreased for couples with paternal pesticide exposure. PMID- 10465179 TI - Zinc supplementation and bone growth in pubertal girls. AB - We supplemented the diets of 47 peripubertal girls with zinc (15 mg/day) or placebo for 6 weeks. Zinc supplementation increased serum zinc. Insulin-like growth factor I and biochemical markers of bone turnover did not change, albeit dietary zinc was below the reference level (in 94% of individuals). PMID- 10465180 TI - Reversibility of vigabratin-induced visual-field defect. AB - We report a girl with epilepsy aged 10 years receiving vigabatrin and complaining of bumping into objects and presenting visual-field constriction, which disappeared after vigabatrin withdrawal. PMID- 10465181 TI - How do Jains get toxoplasma infection? AB - Toxoplasma infection is as prevalent among Jains as among other religious groups in the same area. Jain laws mean that exposure to Toxoplasma gondii via well known routes of infection is unlikely; drinking water may be the vehicle of infection. PMID- 10465183 TI - Chocolate as a source of tea flavonoids. AB - The antioxidant catechin content of chocolate is four times that of tea. Chocolate contributed 20% of the catechin intake in a representative sample of the Dutch population, and tea contributes 55%. Epidemiological assessments of health effects of tea should include other foods that are sources of catechins. PMID- 10465182 TI - Hormone replacement therapy and sensitive C-reactive protein concentrations in women with type-2 diabetes. AB - C-reactive protein concentrations as a marker of inflammation predicts vascular risk and is raised in type-2 diabetes. In a 6-month double-blind placebo controlled trial, a combination of transdermal oestradiol 80 microg with continuous oral norethisterone 1 mg significantly reduced C-reactive protein concentrations in postmenopausal women with type-2 diabetes. PMID- 10465184 TI - Another paediatric cardiac inquiry for the UK. PMID- 10465185 TI - Pharmaceutical companies target plant products for drugs of the future. PMID- 10465186 TI - Mariano Barbacid: a basic scientist branches out. PMID- 10465187 TI - Role of pharmacies in Chinese world of health care. PMID- 10465188 TI - Soldiers claim ill health after contact with contaminated soil in Croatia. PMID- 10465189 TI - Acute respiratory distress syndrome. AB - Outcome in acute respiratory distress syndrome (ARDS) is influenced by a number of factors, including the nature of the precipitating condition and the extent to which multiorgan failure ensues. Most studies of potential therapeutic interventions have been unsuccessful due to the enrollment of limited numbers of patients with a wide variety of pathologies of varying severity. Moreover, the value of initiating single-agent interventions at varying time points in what is an evolving and complex inflammatory process must be questioned. Mortality may therefore represent an inappropriate end-point for clinical trials, which are increasingly focusing on ventilator-free days. Despite these uncertainties, survival appears to be improving, possibly due to the application of supportive techniques in a protocol-driven fashion to patients in whom the underlying condition has been rigorously treated. PMID- 10465190 TI - Manfred Freiherr von Richthofen's medical record--was the "Red Baron" fit to fly? PMID- 10465191 TI - Lactate is an unreliable indicator of tissue hypoxia in injury or sepsis. AB - High blood lactate concentration (hyperlactacidaemia) in trauma or sepsis is thought to indicate tissue hypoxia and anaerobic glycolysis even when blood pressure, cardiac output, and urine output are within clinically acceptable ranges. However, mechanisms of lactate generation by well-oxygenated tissues have received little attention. Within cells, oxidative and glycolytic energy production can proceed in separate, independent compartments. In skeletal muscle and other tissues, aerobic glycolysis is linked to ATP provision for the Na+-K+ pump, the activity of which is stimulated by epinephrine. In injured patients, hypokalaemia may reflect increased Na+,K+-ATPase activity. We propose that increased blood lactate often reflects increased aerobic glycolysis in skeletal muscle secondary to epinephrine-stimulated Na+,K+-ATPase activity and not anaerobic glycolysis due to hypoperfusion. The hypothesis explains why hyperlactacidaemia often neither correlates with traditional indicators of perfusion nor diminishes with increased oxygen delivery. When other variables have returned to normal, continued attempts at resuscitation based on elevated blood lactate may lead to unnecessary use of blood transfusion and inotropic agents in an effort to increase oxygen delivery and lactate clearance. PMID- 10465192 TI - Screening for ovarian cancer. PMID- 10465193 TI - Screening for ovarian cancer. PMID- 10465194 TI - Screening for ovarian cancer. PMID- 10465195 TI - Drug treatments of levodopa-induced dyskinesias. PMID- 10465196 TI - Infertility, preterm birth, and bacterial vaginosis. PMID- 10465197 TI - Interferon beta in multiple sclerosis. PMID- 10465198 TI - Interferon beta in multiple sclerosis. PMID- 10465199 TI - Future of minimally invasive surgery. PMID- 10465200 TI - "Doctor, is wine good for my heart?". PMID- 10465201 TI - "Doctor, is wine good for my heart?". PMID- 10465202 TI - Multidrug-resistant Salmonella typhi in India. PMID- 10465203 TI - Oral care and pneumonia. Oral Care Working Group. PMID- 10465204 TI - Asthma and diabetes. PMID- 10465205 TI - Asthma and diabetes. PMID- 10465206 TI - Effect of water on arteries. PMID- 10465207 TI - Geography of biomedical publications. PMID- 10465208 TI - Geography of biomedical publications. PMID- 10465209 TI - Keeping relatives informed by the internet. PMID- 10465210 TI - How can we help Iraqi people. PMID- 10465211 TI - Informed consent: what did the doctor say? PMID- 10465212 TI - Rise and fall of modern medicine. PMID- 10465213 TI - The Nobel chronicles. 1966: Francis Peyton Rous (1879-1970) and Charles Brenton Huggins (1901-97) PMID- 10465214 TI - Gangsters meet mainstream psychiatry. PMID- 10465215 TI - Inexorably bound in an identical relationship. PMID- 10465216 TI - Provocation-neutralization in the treatment of food allergy. PMID- 10465217 TI - The family physician as a hospitalist. PMID- 10465218 TI - Acute HIV-1 infection: early identification and treatment. PMID- 10465219 TI - The conundrum of early HIV infection. PMID- 10465220 TI - Current trends in cervical ripening and labor induction. PMID- 10465221 TI - Management of diabetic ketoacidosis. AB - Diabetic ketoacidosis is an emergency medical condition that can be life threatening if not treated properly. The incidence of this condition may be increasing, and a 1 to 2 percent mortality rate has stubbornly persisted since the 1970s. Diabetic ketoacidosis occurs most often in patients with type 1 diabetes (formerly called insulin-dependent diabetes mellitus); however, its occurrence in patients with type 2 diabetes (formerly called non-insulin dependent diabetes mellitus), particularly obese black patients, is not as rare as was once thought. The management of patients with diabetic ketoacidosis includes obtaining a thorough but rapid history and performing a physical examination in an attempt to identify possible precipitating factors. The major treatment of this condition is initial rehydration (using isotonic saline) with subsequent potassium replacement and low-dose insulin therapy. The use of bicarbonate is not recommended in most patients. Cerebral edema, one of the most dire complications of diabetic ketoacidosis, occurs more commonly in children and adolescents than in adults. Continuous follow-up of patients using treatment algorithms and flow sheets can help to minimize adverse outcomes. Preventive measures include patient education and instructions for the patient to contact the physician early during an illness. PMID- 10465222 TI - Case studies in international travelers. AB - Family physicians should be alert for unusual diseases in patients who are returning from foreign travel. Malaria is a potentially fatal disease that can be acquired by travelers to certain areas of the world, primarily developing nations. Transmitted through the bite of the Anopheles mosquito, malaria usually presents with fever and a vague systemic illness. The disease is diagnosed by demonstration of Plasmodium organisms on a specially prepared blood film. Travelers can also acquire amebic infections, which may cause dysentery or, in some instances, liver abscess. Amebiasis is diagnosed by finding Entamoeba histolytica cysts or trophozoites in the stool. Invasive amebic infections are generally treated with metronidazole followed by iodoquinol or paromomycin. Cutaneous larva migrans is acquired by skin contact with hookworm larvae in the soil. The infection is characterized by the development of itchy papules followed by serpiginous or linear streaks. Cutaneous larva migrans is treated with invermectin or albendazole. Case studies are presented. PMID- 10465224 TI - Primary dysmenorrhea. AB - Primary dysmenorrhea is defined as cramping pain in the lower abdomen occurring just before or during menstruation, in the absence of other diseases such as endometriosis. Prevalence rates are as high as 90 percent. Initial presentation of primary dysmenorrhea typically occurs in adolescence. It is a common cause of absenteeism and reduced quality of life in women. The problem is often underdiagnosed and undertreated. Women with primary dysmenorrhea have increased production of endometrial prostaglandin, resulting in increased uterine tone and stronger, more frequent uterine contractions. A diagnostic evaluation is unnecessary in patients with typical symptoms and no risk factors for secondary causes. Nonsteroidal anti-inflammatory medications are the mainstay of treatment, with the addition of oral contraceptive pills when necessary. About 10 percent of affected women do not respond to these measures. It is important to consider secondary causes of dysmenorrhea in women who do not respond to initial treatment. Many alternative treatments (ranging from acupuncture to laparoscopic surgery) have been studied, but the supporting studies are small, with limited long-term follow-up. PMID- 10465223 TI - Current trends in cervical ripening and labor induction. AB - Labor is induced in more than 13 percent of deliveries in the United States. Postdate pregnancy is the most common indication. Oxytocin is the drug of choice for labor induction when the cervical examination shows that the cervix is favorable. The use of this agent requires experience and vigilant observation for uterine hyperstimulation, hypertonus or maternal fluid overload. In a patient whose cervix is unfavorable, the use of prostaglandin analogs for cervical ripening markedly enhances the success of inductions. Misoprostol, a prostaglandin E1 analog marketed as a gastrointestinal mucosal protective agent, is safe, efficacious and inexpensive for use in cervical ripening and labor induction. Further studies will better delineate its optimal use. Family physicians need to be familiar with the various methods of cervical ripening and labor induction. PMID- 10465225 TI - Reactive arthritis (Reiter's syndrome). AB - Reactive arthritis, also called Reiter's syndrome, is the most common type of inflammatory polyarthritis in young men. It is sometimes the first manifestation of human immunodeficiency virus infection. An HLA-B27 genotype is a predisposing factor in over two thirds of patients with reactive arthritis. The syndrome most frequently follows genitourinary infection with Chlamydia trachomatis, but other organisms have also been implicated. Treatment with doxycycline or its analogs sometimes shortens the course or aborts the onset of the arthritis. Reactive arthritis may also follow enteric infections with some strains of Salmonella or Shigella, but use of antibiotics in these patients has not been shown to be effective. Reactive arthritis should always be considered in young men who present with polyarthritis. Symptoms may persist for long periods and may, in some cases, cause long-term disability. Initial treatment consists of high doses of potent nonsteroidal anti-inflammatory drugs. Patients with large-joint involvement may also benefit from intra-articular corticosteroid injection. PMID- 10465226 TI - Endogenous endophthalmitis: case report and brief review. AB - Endogenous endophthalmitis is a potentially blinding ocular infection resulting from hematogenous spread from a remote primary source. The condition is relatively rare but may become more common as the number of chronically debilitated patients and the use of invasive procedures increase. Many etiologic organisms (gram-positive, gram-negative and fungal) have been reported to cause endogenous endophthalmitis. Risk factors are well defined and include most reasons for immune suppression. A high clinical suspicion is needed for early diagnosis and treatment. Early intravenous antibiotic therapy remains the cornerstone of treatment. The roles of intravitreal antibiotics and vitrectomy are evolving and may become more widely accepted as therapeutic modalities. The authors report a case of endogenous endophthalmitis and provide a brief review of the literature. PMID- 10465227 TI - Post-traumatic stress reactions following motor vehicle accidents. AB - Despite improvements in road conditions, vehicle safety and driver education, over 3 million persons are injured in motor vehicle accidents each year. Many of these persons develop post-traumatic stress symptoms that can become chronic. Patients with post-traumatic stress disorder experience disabling memories and anxiety related to the traumatic event. Early identification of these patients is critical to allow for intervention and prevent greater impairment and restriction. The family physician is in an ideal position to identify, treat or refer patients with traumatic responses to traffic accidents. The physician's awareness of patient characteristics and pre-accident functioning allows him or her to critically evaluate symptoms that may begin to interfere with the resumption of daily activities. PMID- 10465228 TI - How to recognize and treat acute HIV syndrome. AB - The diagnosis of acute human immunodeficiency virus (HIV) syndrome requires a high index of suspicion and proper laboratory testing. Patients with the syndrome may have fever, fatigue, rash, pharyngitis or other symptoms. Primary HIV infection should be considered in any patient with possible HIV exposure who presents with fever of unknown cause. The diagnosis is based on a positive HIV-1 RNA level (more than 50,000 copies per mL) in the absence of a positive enzyme linked immunosorbent antibody assay (ELISA) and confirmatory Western blot antibody test for HIV. Early diagnosis permits patient education as well as treatment that may delay disease progression. Triple-combination antiretroviral therapy should be started immediately and continued indefinitely. Compliance with medication regimens is essential to maximize benefit and discourage the development of viral resistance. PMID- 10465229 TI - Peyronie's disease: current management. AB - Peyronie's disease is an acquired inflammatory condition of the penis associated with penile curvature and,in some cases, pain. It primarily affects men between 45 and 60 years of age, although an age range of 18 to 80 years has been reported. If left untreated, Peyronie's disease may cause fibrotic, nonexpansile thickening of relatively discrete areas of the corpora tunica, typically resulting in focal bend, pain or other functional or structural abnormalities of the erect penis. Many cases resolve without treatment. Medical therapies, including antioxidants (such as vitamin E and potassium aminobenzoate) and corticosteroids injected directly into the plaque, lack adequate scientific support. Surgery remains a mainstay when conservative measures fail. PMID- 10465230 TI - Heart murmurs in pediatric patients: when do you refer? AB - Many normal children have heart murmurs, but most children do not have heart disease. An appropriate history and a properly conducted physical examination can identify children at increased risk for significant heart disease. Pathologic causes of systolic murmurs include atrial and ventricular septal defects, pulmonary or aortic outflow tract abnormalities, and patent ductus arteriosus. An atrial septal defect is often confused with a functional murmur, but the conditions can usually be differentiated based on specific physical findings. Characteristics of pathologic murmurs include a sound level of grade 3 or louder, a diastolic murmur or an increase in intensity when the patient is standing. Most children with any of these findings should be referred to a pediatric cardiologist. PMID- 10465231 TI - Soft tissue sarcomas: integrating primary care recognition with tertiary care center treatment. AB - Soft tissue sarcomas account for fewer than 1 percent of malignancies diagnosed annually in the United States. These tumors usually present as an asymptomatic mass. Any lesion larger than 5 cm in diameter should be considered suspicious. Radiographs should be obtained as the initial step in assessing a suspicious lesion. Magnetic resonance imaging has become the preferred diagnostic examination for tumors involving the extremities, and computed tomographic scanning may be the best technique for imaging lesions in the thoracic, abdominal, and head and neck areas. In general, the patient with a suspicious soft tissue mass located in a surgically difficult area should be referred to a regional center for biopsy and multidisciplinary consultation before resection is attempted. Careful preoperative planning is necessary for a good outcome. The prognosis for the patient with a soft tissue sarcoma is primarily determined by the grade, size and depth of the tumor and the presence of tumor at the surgical margins. PMID- 10465232 TI - Protecting the health and safety of working teenagers. AB - More than one third of high school students work during the school year, and many more are employed during the summer months. Teenage workers face a variety of health and safety hazards. Occupational injury and illness are largely preventable, and family physicians can play a crucial role in this prevention effort by advising adolescents about common workplace dangers. Physicians who sign work permits and provide ongoing health care to teenagers should counsel them and their parents or guardians about the benefits and risks of work and discuss the regulations governing jobs that are prohibited for adolescents, work hours, protective measures and workers' compensation benefits. PMID- 10465233 TI - Photo quiz. Food intolerance. PMID- 10465234 TI - Female circumcision. PMID- 10465235 TI - Adolescent health. American Academy of Family Physicians. PMID- 10465236 TI - AAP issues guidelines for urinary tract infections in infants and toddlers. American Academy of Pediatrics. PMID- 10465237 TI - Eleven national medical associations join to prevent pneumonia. PMID- 10465238 TI - Availability of updated HIV treatment guidelines. PMID- 10465239 TI - Effect of methionine supplementation on endothelial function, plasma homocysteine, and lipid peroxidation. AB - BACKGROUND: Acetaminophen (paracetamol) poisoning is a major source of morbidity and mortality. It has been proposed that methionine be incorporated into acetaminophen tablets routinely as a protective mechanism. Methionine has been shown to be effective in the treatment of acetaminophen toxicity and a combination preparation of acetaminophen and methionine may prevent toxicity. However, there has been some concern that chronic methionine supplementation may be associated with vascular disease. The aim of the study was to investigate if methionine supplementation causes changes in endothelial function, plasma homocysteine, or lipid peroxidation which may be associated with atherosclerosis. METHODS: Sixteen healthy volunteers were studied. Forearm blood flow in response to local intra-arterial infusion of acetylcholine to assess endothelium-dependent vasodilatation and sodium nitroprusside to assess endothelium-independent vasodilatation was measured by venous occlusion plethysmography. Plasma homocysteine and lipid peroxidation, measured as thiobarbituric acid reactive substances, were measured using high-performance liquid chromatography. Forearm vascular responses, plasma homocysteine concentrations, and thiobarbituric acid reactive substances were measured at baseline and following methionine supplementation. RESULTS: There was no significant difference in endothelial dependent vascular responses after acute (methionine 250 mg orally, p > 0.05), 1 month of low-dose (methionine 250 mg daily, p > 0.05), or 1 week of high-dose (methionine 100 mg/kg daily, p > 0.05) methionine administration. There was no significant difference in plasma homocysteine concentrations after acute (p > 0.05) or 1 month of low-dose (p > 0.05) methionine administration. However, 1 week of high-dose methionine (100 mg/kg) administration daily significantly increased homocysteine concentrations (p < 0.0015). Thiobarbituric acid reactive substances were unchanged during the period of study (p > 0.05). CONCLUSIONS: Methionine supplementation does not impair endothelial-dependent vascular responses in healthy volunteers. Although high-dose methionine administration causes elevation of plasma homocysteine concentrations, doses similar to those used in combination preparations with acetaminophen do not affect plasma homocysteine concentrations. PMID- 10465240 TI - Opiate detection in saliva and urine--a prospective comparison by gas chromatography-mass spectrometry. AB - There is an increasing interest in saliva as an alternative analytic body fluid. OBJECTIVE: This study sought to determine the correlation of opiates analyzed in saliva and corresponding urine. METHODS: A total of 130 adequate and 24 inadequate samples were collected from patients participating in drug withdrawal therapy. To obtain saliva from the oral cavity, a newly developed collection device (Clin Rep), consisting of a treated cotton roll and a centrifugation vial with a filter inset, was tested. For the preparation of a purified solution of urine, liquid extraction was used. Solid phase extraction was utilized to prepare the saliva samples. For the detection by gas chromatography-mass spectrometry, an appropriate derivatization was necessary using N-methyl-N (trimethylsilyl)trifluoroacetamide. The retention times were compared with defined standard solutions. The obtained mass spectra showed a characteristic fragmenting pattern and offered a reliable identification. RESULTS: The concordance of the analytic results of the saliva samples with urine was 93% for a decision limit of 100 ng/mL and 98% for a decision limit of 300 ng/mL (DHHS opiate cutoff) in urine. CONCLUSIONS: Saliva, of adequate amount in 85% of samples, may be appropriate for analysis of drugs of abuse. PMID- 10465241 TI - Loxosceles deserta spider venom induces NF-kappaB-dependent chemokine production by endothelial cells. AB - BACKGROUND: Loxosceles spider evenomation in man frequently results in disfiguring necrotic skin lesions. Recent studies suggest that several proinflammatory mediators participate in lesion development. We have observed that Loxosceles deserta venom induces production of the chemokines interleukin-8, growth-related oncogene alpha, and monocyte chemoattractant protein-I by human umbilical vein endothelial cells. Members of the Rel/Nuclear factor (NF)-kappaB family of transcription factors are important regulators of many genes involved in immune and inflammatory responses. We hypothesized that Loxosceles-venom induced chemokine expression in human umbilical vein endothelial cells is mediated by NF-kappaB. METHODS: Human umbilical vein endothelial cell monolayers were exposed to activating concentrations of Loxosceles deserta venom. Nuclear extracts of these monolayers were analyzed by electrophoretic mobility shift assay. A direct cause and effect linkage between NF-kappaB activation and chemokine expression by Loxosceles venom was established through examination of the effect of SN50 on interleukin-8 and monocyte chemoattractant protein-1 production using a whole-cell enzyme immunoassay. SN50 is a cell-permeable peptide that specifically blocks cytosolic to nuclear translocation of NF-kappaB. Furthermore, the venom-induced synthesis of chemokine mRNAs was investigated by RNase protection assays. RESULTS: Loxosceles deserta venom induces the activation of NF-kappaB in human umbilical vein endothelial cells. Antibodies to p50 and p65, but not to p52, c-Rel, or Rel B, induce supershifts of the DNA-protein complexes formed by oligonucleotide probes and nuclear extracts from venom activated human umbilical vein endothelial cells. SN50 peptide inhibits NF-kappaB translocation and interleukin-8 and monocyte chemoattractant protein-1 production in activated human umbilical vein endothelial cells. CONCLUSIONS: Loxosceles deserta venom induces synthesis of interleukin8 and monocyte chemoattractant protein-1 mRNAs in human umbilical vein endothelial cells. The expression of chemokines occurs via an NF-kappaB-dependent pathway. PMID- 10465242 TI - A retrospective analysis of 96 "asp" (Megalopyge opercularis) envenomations in Central Texas during 1996. AB - BACKGROUND: The most frequently reported caterpillar envenomation in Central Texas is by the puss caterpillar or "asp," Megalopyge opercularis. This caterpillar is described by patients and physicians as inflicting intense radiating pain. The intensity of symptoms may be underestimated leading to undertreatment. Adequate treatment protocols have been lacking and those in use are not very successful. We present a retrospective study of patients who were stung and contacted the Central Texas Poison Center. METHODS: All human exposures to asp stings reported to the Central Texas Poison Center during 1996 were included. Inclusion criteria consisted of all cases documented as an asp envenomation by the specialists in poison information. Characterization of symptoms and treatment used were evaluated. RESULTS: There were 96 exposures to asps reported. Ninety-five of the patients experienced local pain with 26 of these reporting intense radiating pain. Forty developed erythema, 27 described edema, and 9 complained of welts/hives. Other symptoms reported included white spots (4), pruritus (3), red streak (2), numbness (2), and individual accounts of chest pain, rash, ecchymosis, tingling, blister, and muscle spasm. There was no treatment modality that promptly relieved pain. DISCUSSION: Although asp envenomations appear to be very common, clinical cases have rarely been documented. This may be due to physicians not recognizing the etiological agent. Pain may be very intense and standard pain management appears to be unsuccessful. CONCLUSION: This study suggests that further examination of treatment modalities may be beneficial in addressing the morbidity of Megalopyge opercularis envenomations. PMID- 10465243 TI - Insulin-glucose as adjunctive therapy for severe calcium channel antagonist poisoning. AB - CASE REPORT: This case series documents the clinical courses of 4 patients after verapamil overdose and 1 patient after amlodipine-atenolol overdose. All subjects had hypodynamic circulatory shock (hypotension, bradycardia, and acidosis) that was not adequately responsive to conventional treatment. After initiation of insulin-dextrose infusion, the hemodynamic status of all 5 patients stabilized and all patients survived. Plasma drug concentrations are reported for all cases and verapamil levels were extremely high in 2 patients (3710 ng/mL and 3980 ng/mL). However, because patients were not treated according to a standard protocol, each patient received variable other supportive measures and inotropic agents, and the infusion rates of insulin were variable among patients. This report provides preliminary evidence toward a larger trial of insulin-dextrose to treat hypodynamic shock from calcium channel blocker overdose. PMID- 10465244 TI - Use of partial liquid ventilation to manage pulmonary complications of acute verapamil-sustained release poisoning. AB - INTRODUCTION: Verapamil is a papaverine-derived calcium channel blocker widely used for the treatment of hypertension and supraventricular tachyarrhythmias. It is one of the leading agents involved in pharmaceutical poisoning-related deaths among adults. CASE REPORT: We report a case of severe sustained-release verapamil poisoning associated with respiratory failure in an adult man who survived after receiving 4 days of partial liquid ventilation as a part of his medical management. PMID- 10465245 TI - Acebutolol-induced ventricular tachycardia reversed with sodium bicarbonate. AB - BACKGROUND: Acebutolol is a unique beta blocker that possesses cardioselectivity, partial agonist activity, and membrane stabilizing activity. Sodium bicarbonate is used to reverse the cardiotoxic effects of other drugs with membrane stabilizing activity. There have been no reported cases of acebutolol-induced ventricular dysrhythmias treated successfully with bolus sodium bicarbonate. CASE PRESENTATION: A 48-year-old man ingested approximately 6.4 g of acebutolol with ethanol (blood ethanol 61 mmol/L). There were no other coingestants identified. One hour after presentation, the patient had a cardiac arrest with the monitor showing ventricular tachycardia. Sodium bicarbonate 50 mEq intravenous push converted the patient to sinus rhythm and the blood pressure improved to 129/90 mm Hg. CONCLUSION: This case demonstrates a temporal relationship between bolus sodium bicarbonate administration and the termination of acebutolol-induced ventricular tachycardia. PMID- 10465246 TI - Hypersensitivity myocarditis associated with ephedra use. AB - BACKGROUND: Ephedrine has previously been described as a causative factor of vasculitis but myocarditis has not yet been associated with either ephedrine or its plant derivative ephedra. CASE REPORT: A 39-year-old African American male with hypertension presented to Rush Presbyterian St. Luke's Medical Center with a 1-month history of progressive dyspnea on exertion, orthopnea, and dependent edema. He was taking Ma Huang (Herbalife) 1-3 tablets twice daily for 3 months along with other vitamin supplements, pravastatin, and furosemide. Physical examination revealed a male in mild respiratory distress. The lung fields had rales at both bases without audible wheezes. Internal jugular venous pulsations were 5 cm above the sternal notch. Medical therapy with intravenous furosemide and oral enalapril was initiated upon admission. Cardiac catheterization with coronary angiography revealed normal coronary arteries, a dilated left ventricle, moderate pulmonary hypertension, and a pulmonary capillary wedge pressure of 34 mm Hg. The patient had right ventricular biopsy performed demonstrating mild myocyte hypertrophy and an infiltrate consisting predominantly of lymphocytes with eosinophils present in significantly increased numbers. Treatment for myocarditis was initiated with azothioprine 200 mg daily and prednisone 60 mg per day with a tapering course over 6 months. Anticoagulation with warfarin and diuretics was initiated and angiotensin-converting enzyme inhibition was continued. Hydralazine was added later. One month into therapy, an echocardiogram demonstrated improved left ventricular function with only mild global hypokinesis. A repeat right ventricular biopsy 2 months after the first admission showed no evidence of myocarditis. At 6 months, left ventricular ejection fraction was normal (EFN 50%) and the patient asymptomatic. CONCLUSION: Ephedra (Ma Huang) is the suspected cause of hypersensitivity myocarditis in this patient due to the temporal course of disease and its propensity to induce vasculitis. PMID- 10465247 TI - Laboratory confirmation of scopolamine co-intoxication in patients using tainted heroin. AB - BACKGROUND: First described in 1995, at least 325 patients with a history of heroin use have since required emergency medical evaluation in several eastern US cities, because of an anticholinergic toxidrome following use of heroin. This co intoxication has been alleged to result from an atropine-like compound. We report the clinical findings and laboratory analysis of one of several individuals who presented to our Emergency Department during this epidemic. CASE REPORT: A 23 year-old male was one of 3 patients brought to the Emergency Department due to agitated behavior after insufflating heroin. Following physical and chemical restraint, vital signs were pulse 134 bpm, BP 160/90 mm Hg, RR 24/min, and T 37.3 degrees C. Physical examination was remarkable for dilated pupils 8-9 mm without nystagmus, along with dry mouth, decreased bowel sounds, and flushed dry skin. A bladder catheter was placed and 500 mL of urine was obtained. Electrocardiogram revealed sinus tachycardia only. Additional sedation was required for 12 hours until normal mental status returned. A small sample of the "heroin" used was obtained and analyzed. Scopolamine was confirmed by gas chromatography-mass spectrometry. Further evidence of scopolamine intoxication was supported by identifying scopolamine in the urine of this patient. CONCLUSION: Gas chromatography-mass spectrometry revealed scopolamine to be the cause of anticholinergic findings in a patient following use of tainted heroin. It is unclear whether scopolamine is an adulterant or contaminant in this heroin. Patients with anticholinergic findings following use of heroin may be co intoxicated with scopolamine. PMID- 10465248 TI - Diverse manifestations of oral methylene chloride poisoning: report of 6 cases. AB - OBJECTIVE: Methylene chloride is a solvent used in domestic and industrial preparations, such as paint removers and degreasing agents. Although it is considered of low toxicity, acute toxic manifestations have been reported following inhalation of methylene chloride, mainly from working in an enclosed environment. Oral ingestion of methylene chloride, however, remains rare and its consequences are less clearly understood. CASE REPORTS: The various clinical manifestations of 6 patients with oral ingestion of methylene chloride are reported. Central nervous system depression, tachypnea, and corrosive gastrointestinal injury were the most common presentations following ingestion. An elevated carboxyhemoglobin level was documented in only 2 patients (35% and 8.4% carboxyhemoglobin, respectively). Due to a frequent misleading history of "chloroform" ingestion and unawareness of probable carboxyhemoglobin production in these patients, carboxyhemoglobin was not routinely measured. Renal failure, hepatic failure, and acute pancreatitis occurred in the 2 most severe cases of methylene chloride ingestion. CONCLUSIONS: Ingestion of methylene chloride can result in diverse manifestations, including a high carboxyhemoglobin level. Corrosive gastrointestinal injury is common in oral poisoning and needs further therapeutic consideration. A high index of suspicion and appropriate laboratory studies are needed in those patients who allegedly ingest "chloroform" but do not present the incriminating solvent to their treating physicians. The presence of an elevated carboxyhemoglobin level suggests the diagnosis of methylene chloride poisoning. Symptomatic and supportive measures remain the mainstay in the treatment of patients with oral methylene chloride poisoning. PMID- 10465249 TI - Effect of charcoal hemoperfusion on clearance of cibenzoline succinate (cifenline) poisoning. AB - BACKGROUND: Management of cibenzoline succinate (cifenline) poisoning by symptomatic treatment is recommended because it has been reported cibenzoline succinate is not effectively removed by hemodialysis. The use of charcoal hemoperfusion for patients with cibenzoline succinate poisoning has not been reported previously. CASE REPORT: An 80-year-old woman with permanent right ventricular pacing was admitted to our hospital with general fatigue and clouding of consciousness. She had been receiving cibenzoline succinate for 1 month. The patient was clearly in shock on admission. Electrocardiogram showed a prolonged QRS and QTc interval and pacing failure. Hemodynamic failure, electrocardiographic abnormalities including pacing failure, and liver and renal dysfunction were compatible with cibenzoline succinate poisoning. Symptomatic treatment was provided. On day 4, charcoal hemoperfusion was initiated because of the development of hypoglycemia accompanied by liver and renal dysfunction. After charcoal hemoperfusion, the hypoglycemia, liver and renal dysfunction, prolonged QRS and QTc interval, and pacing threshold resolved. She was discharged on digoxin. We present a case of cibenzoline succinate poisoning treated with charcoal hemoperfusion which resulted in a rapid reduction of cibenzoline plasma concentrations with a significant clinical improvement. PMID- 10465250 TI - Rabbit syndrome following phenol ingestion. AB - CASE REPORT: An elderly Japanese woman ingested a massive quantity of phenol in a suicide attempt. She was admitted to the Emergency Department in respiratory arrest and deep coma. Duodenogastritis was evident endoscopically. With the return of spontaneous respiration and consciousness, fine, rapid rhythmic perioral movements developed together with Parkinsonian findings. The abnormal movements were aggravated by administration of a neuroleptic and ameliorated by discontinuing the drug; they disappeared completely by hospital day 15. In addition to neuroleptic drugs, phenol intoxication may cause the rabbit syndrome by inducing cholinergic dominance with relative dopamine hypofunction in the central nervous system. PMID- 10465251 TI - Amitraz poisoning in humans. PMID- 10465252 TI - Urine cannabinoids and usage of legal hemp products. PMID- 10465253 TI - Health hazards due to clenbuterol residues in food. PMID- 10465254 TI - Plantlore revisited. PMID- 10465255 TI - Insulin-like growth factor I--a prototypic peripheral-paracrine hormone? PMID- 10465256 TI - Somatotroph and lactotroph changes in the adenohypophyses of mice with disrupted insulin-like growth factor I gene. AB - Pituitary is influenced by circulating and locally produced insulin-like growth factor I (IGF-I). To further elucidate the role of pituitary IGF-I, we compared pituitary morphology of homozygous (IgfI-/-), heterozygous (IgfI+/+), and wild type (IgfI+/+) fetal and adult mice using light microscopy, immunocytochemistry, in situ hybridization and electron microscopy. In pituitaries of Igf1-/- and Igf1+/- fetal mice (day 18.5) GH RNA signal was decreased. In Igf1-/- adult females, GH cells were significantly diminished in size; GH RNA signal was stronger in Igf1-/- mice compared with IgfI+/+ mice, and the somatotrophs had ultrastructural features of stimulation. The number of PRL cells and PRL hybridization signal were significantly decreased, however plasma PRL levels were elevated in both genders. No changes in other cell types in Igf1-/- mice, and no alterations in Igf1+/- mice were evident. IGF-I treatment for 2 weeks of Igf1-/- mice increased significantly body weights, decreased GH hybridization signal, and had no effect on PRL cells, or PRL plasma levels, whereas in IgfI+/+ mice, PRL RNA signal and PRL plasma levels were markedly increased. In conclusion, IGF-I plays no role in differentiation of pituitary cells, affects the size of somatotrophs in females, and is a stimulator of lactotrophs in both genders. PMID- 10465257 TI - Interleukin-6 is a needed proinflammatory cytokine in the prolonged neural activity and transcriptional activation of corticotropin-releasing factor during endotoxemia. AB - Interleukin-6 (IL-6) is a proinflammatory cytokine that plays multiple roles in the central nervous system during infections and injuries. Although this molecule is capable of stimulating the release of ACTH and glucocorticoids, it has been demonstrated that a single injection of IL-6 fails to activate the paraventricular nucleus (PVN) neurons that control the hypothalamic-pituitary adrenal axis. The observation that IL-6 receptor (IL-6R) is up-regulated in the brain during endotoxemia led us to hypothesize that prior induction of IL-6R synthesis could amplify the effect of circulating IL-6 on the neuroendocrine response. Rats received a first iv injection of either bacterial lipopolysaccharide (LPS; 5 microg) or vehicle solution. After a 6-h waiting period, they received a second iv injection of either recombinant rat IL-6 or vehicle solution and were killed 1 h thereafter. Using in situ hybridization, we observed that IL-6R was barely expressed in the PVN under basal conditions, but was rapidly produced in response to LPS. IL-6 itself was also able to induce the synthesis of its own receptor along cerebral blood vessels, and this effect extended to several parenchymal structures, including the PVN, when the cytokine was administrated after LPS. In agreement with our hypothesis, we found that IL-6 injected in LPS-pretreated rats stimulated PVN neurons, as revealed by the expression of CRF primary transcript and c-fos messenger RNA, an immediate early gene used as a marker of cellular activation. A significant increase in plasma corticosterone levels was also found in animals that received iv IL-6 injection after being pretreated 6 h before with the very low dose of LPS. The fact that IL 6 alone or injected after LPS treatment was unable to induce cyclooxygenase-2 synthesis is an argument in favor of a PG-independent mechanism. The relative contribution of IL-6 in stimulating CRF expression in the PVN and neural activity throughout the brain during endotoxemia was also investigated in IL-6-deficient mice after an ip injection of LPS. The endotoxin induced similar c-fos and CRF expression patterns in knockout and wild-type mice, but the expression levels were generally higher and/or lasted longer in wild-type animals. Taken together, physiological changes that may include the induction of IL-6R synthesis seem to be necessary for IL-6 to activate PVN neurons. Moreover, although IL-6 does not appear essential during the early phases of endotoxemia, this cytokine is required during the later phases to prolong the activation of neural cells throughout the brain and to maintain CRF expression in the PVN neurons that control the hypothalamic-pituitary-adrenal axis. PMID- 10465258 TI - Steroid regulation of human placental integrins: suppression of alpha2 integrin expression in cytotrophoblasts by glucocorticoids. AB - Maintenance of uterine-placental attachment during human pregnancy may depend at least partly on adhesive interactions between cytotrophoblasts and their extracellular matrix (ECM). Such interactions are often mediated by integrins, signal-transducing heterodimeric transmembrane glycoproteins. We previously showed that glucocorticoid (GC) suppressed the expression of collagen and laminin in human placenta; here we show that GC also modulates the expression by human cytotrophoblasts of the integrin subunits alpha2 and beta1, components of a known receptor for these ECM ligands. Cytotrophoblasts were isolated from human term placentas, cultured up to 4 days in the presence of 0-1000 nM dexamethasone (DEX), and assayed for 1) integrin messenger RNA (mRNA) levels by Northern hybridization, 2) integrin subunit synthesis after [35S]methionine labeling, or 3) cell surface integrin levels after 125I labeling by lactoperoxidase. In four independent experiments, 100 nM DEX reduced mRNA levels for integrin alpha2 to 6+/-1% of the control value. This effect was similar between 1-4 days of treatment and was dose dependent between 1-1000 nM DEX. Cortisol treatment (100 nM) inhibited levels of integrin alpha2 mRNA, but 100 nM testosterone, estradiol, and progesterone were less effective, suggesting that this response was specific to GC. In immunoprecipitation studies, treatment of cytotrophoblasts with 100 nM DEX for 2 days reduced the rates of synthesis of the alpha2 integrin subunit as well as its expression on the cell surface to 1-10% of control levels. DEX effects on the beta1 integrin subunit were less dramatic. DEX reduced beta1 mRNA levels to only 69+/-8% of control levels, a smaller reduction compared with effects on alpha2 integrin mRNA. DEX inhibited beta1 protein synthesis and cell surface expression to 60-70% of control levels. In all experiments, DEX had no effect on total protein synthesis. Thus, our results demonstrate that GC treatment specifically and markedly down-regulates expression of alpha2 integrin subunit by human cytotrophoblasts. This finding is consistent with the concept that uterine-placental adherence across gestation may be regulated by coordinate effects on ECM ligands and cellular adhesion receptors. PMID- 10465259 TI - The 20-kilodalton (kDa) human growth hormone (hGH) differs from the 22-kDa hGH in the effect on the human prolactin receptor. AB - Previously we have demonstrated that 20-kDa human GH (20K-hGH) is a full agonist for hGH receptor (hGHR) even though its complex formation with hGHR and hGH binding protein differs from that of 22-kDa human GH (22K-hGH). In this study, we focused on the effect of 20K-hGH on human PRL receptor (hPRLR). To elucidate the effects of 20K-hGH on hPRLR and compare them with those of 22K-hGH, we prepared two cells stably expressing full-length hPRLR, Ba/F3-hPRLR and CHO-hPRLR. In the proliferation of Ba/F3-hPRLR cells, which can grow in a dose-response to lactogenic hormones, both 20K- and 22K-hGH exhibited bell-shaped curves in the absence of exogenous zinc ion (Zn2+); however, the curve of 20K-hGH was shifted to a 10-fold higher concentration than that of 22K-hGH in view of EC50 value (the EC50 of 20K- and 22K-hGH were 15 nM and 1.5 nM, respectively). Addition of Zn2+ up to 25 microM increased the activities of both 20K- and 22K-hGH; however, the enhancement by Zn2+ was greater in 20K-hGH than in 22K-hGH, thereby the activities of both hGH isoforms reached the same level at 25 microM Zn2+. Nevertheless, in the presence of 0.25-1 microM free Zn2+, which is equal in human serum, the activity of 20K-hGH was still lower than that of 22K-hGH. The modest effect of 20K-hGH on activating hPRLR in the absence of Zn2+ was confirmed in the rat serine protease inhibitor 2.1 (Spi2.1) gene promoter activation and JAK2/Stat5 tyrosine phosphorylation in CHO-hPRLR. In addition, in human breast cancer cell T-47D, 20K-hGH was proved to stimulate Stat5 tyrosine phosphorylation to much lower degree than 22K-hGH via not hGHR but hPRLR. Taken together, our data suggest that 20K-hGH may be a weaker agonist for hPRLR than 22K-hGH in the human body; therefore 20K-hGH may alleviate the hPRLR-mediated side-effects such as breast cancer when administered to human body. PMID- 10465261 TI - Effect of estrogen agonists and antagonists on induction of progesterone receptor in a rat hypothalamic cell line. AB - Estrogen is essential in the hypothalamus for the central regulation of reproduction. To understand the molecular mechanism(s) of estrogen action in the hypothalamus, immortalized rat embryonic hypothalamic cell lines were characterized for steroid receptors and subcloned. Scatchard analysis of the D12 subclone demonstrated one high affinity estrogen receptor-binding site (Kd = 31.3+/-1.9 pM) with a Bmax of 30.8+/-0.8 fmol/mg. Estrogen receptor-alpha protein was identified by Western blot and gel shift analyses. Treatment with estradiol (48 h) stimulated progesterone receptor (PR) messenger RNA expression and binding to [3H]R5020, a synthetic progestin. Because the agonist or antagonist activity of estrogen mimetics can be cell type dependent, the activities of various estrogen mimetics were determined in D12 cells. ICI 182,780 (IC50 = 0.63 nM), raloxifene (IC50 = 1 nM), enclomiphene (IC50 = 77 nM), and tamoxifen (IC50 = 174 nM) inhibited the induction of PR by estradiol, and none of these compounds significantly stimulated PR when given alone. In contrast, 17alpha-ethynyl estradiol (EC50 = 0.014 nM), zuclomiphene (EC50 = 100 nM), and genistein (EC50 = 17.5 nM) functioned as estrogen agonists in these cells. In addition, the estrogen-induced progesterone receptor activated a progesterone response element reporter construct in response to progestins. Thus, the D12 rat hypothalamic cell line provides a useful model for characterizing tissue-selective estrogenic compounds, identifying estrogen- and progesterone-regulated hypothalamic genes, and understanding the molecular mechanisms of steroid action in various physiological processes mediated by the hypothalamus. PMID- 10465260 TI - Glucagon-like peptide 1 elevates cytosolic calcium in pancreatic beta-cells independently of protein kinase A. AB - Glucagon-like peptide 1 (7-36)amide (GLP-1) is an insulinotropic intestinal peptide hormone with a potential role as antidiabetogenic therapeutic agent. It mediates a potentiation of glucose-induced insulin secretion, by activation of adenylate cyclase and subsequent elevation of cytosolic free calcium, [Ca2+]cyt. We investigated the role of protein kinase A (PKA) in GLP-1 signal transduction, using isolated mouse islets as well as the differentiated beta-cell line INS-1. Two specific inhibitors of PKA, (Rp)-adenosine cyclic 3',5'-phosporothioate (Rp cAMPS, up to 3 mM) and KT5720 (up to 10 microM), did not inhibit the GLP-1 induced [Ca2+]cyt elevation. Another PKA inhibitor, H-89, reduced the [Ca2+]cyt elevation only when applied at high concentrations (10-40 microM), higher than sufficient for PKA inhibition in many cell types. Furthermore, at these concentrations, H-89 also inhibited presumably PKA-independent processes such as glucose-induced [Ca2+]cyt elevations and intracellular calcium storage. This suggests a PKA-independent action of H-89. Similarly to H-89, the potent but unselective protein kinase inhibitor staurosporine inhibited the GLP-1-induced [Ca2+]cyt elevation only at high concentrations, at which it also inhibited glucose-induced [Ca2+]cyt elevations. The same observations as with GLP-1 were made when adenylate cyclase was stimulated with forskolin, for selective examination of signal transduction downstream of receptor and G protein. Our results suggest that the GLP-1-induced [Ca2+]cyt elevation is mediated independently of PKA and thus belongs to the yet-little-characterized ensemble of effects that are mediated by binding of cAMP to other target proteins. PMID- 10465262 TI - Spermatogenesis without gonadotropins: maintenance has a lower testosterone threshold than initiation. AB - We showed previously that testosterone (T) alone could induce spermatogenesis and produce normally fertile spermatozoa in the absence of circulating gonadotropins. These studies used the hpg mouse, which is characterized by a congenital gonadotrophin deficiency due to a major deletion in the GnRH gene. Administering T by a subdermal implant of a SILASTIC brand tube impregnated with crystalline T showed that the androgenic requirement for full induction of spermatogenesis was a 1-cm length implant. Using this unique model of spermatogenesis without gonadotropins, we have now investigated the quantitative requirement for androgens to maintain spermatogenesis by testing the hypothesis that the androgenic threshold required for induction and maintenance of spermatogenesis are the same. Spermatogenesis was induced in homozygous hpg mice by T administration for 6 weeks. The first experiment determined the time-course of the regression of spermatogenesis after removal of the T-impregnated SILASTIC brand implant. Elongated spermatids were absent by 3 weeks and testicular weight regression was maximal by 4 weeks after androgen withdrawal. The second experiment examined the effects on maintenance of spermatogenesis of reducing the T dose. After full induction of spermatogenesis in homozygous hpg mice, the T implants were replaced with a range of smaller size T-impregnated SILASTIC brand implants for a further 4 weeks. All androgen-sensitive end-points (testis weight, tubular, and luminal diameters, round spermatids) were fully maintained with T implants of 0.06 cm and elongated spermatids with T implants of 0.25 cm. A further experiment showed that at very low T doses (0.06, 0.125 cm) the T effects observed at 4 weeks were maintained at 6 and 11 weeks duration. We conclude that the androgenic threshold to maintain spermatogenesis in the mouse is an order of magnitude lower than the threshold required for inducing spermatogenesis. This distinction suggests that the mechanism of action of testosterone in inducing spermatogenesis may involve regulation of a genetic switch to complete meiosis, whereas the maintenance involves a different locus of action. These findings suggest that further studies of androgen-dependent meiotic genes may be central to understanding the regulation and molecular basis of androgen-driven induction and maintenance of spermatogenesis. PMID- 10465263 TI - Reduced growth hormone receptor (GHR) messenger ribonucleic acid in liver of periparturient cattle is caused by a specific down-regulation of GHR 1A that is associated with decreased insulin-like growth factor I. AB - GH receptor (GHR) messenger RNA (mRNA) is transcribed from at least three different promoters within the liver of cattle. The first promoter (P1) is liver specific and alternatively splices exon 1A onto the GHR mRNA (GHR 1A mRNA). The second and third promoters (P2 and P3) have constitutive activity in many tissues and alternatively splice exons 1B and 1C onto the GHR mRNA (GHR 1B and GHR 1C mRNA). The total amount of GHR in the liver partially determines liver insulin like growth factor I (IGF-I) synthesis in response to GH. Two studies were conducted to characterize the changes in GHR 1A mRNA, alternatively spliced GHR mRNA, and IGF-I mRNA during late pregnancy and early lactation in dairy cattle. Liver RNA was isolated from pregnant Holstein cattle (Bos taurus) on days -14, 0, and 21 relative to parturition (study 1) or days -14, 0, 15, 30, 60, and 90 relative to parturition (study 2). Ribonuclease protection assays were used to quantify total GHR (all GHR variants) as well as liver-specific GHR 1A and alternatively spliced GHR mRNA. Likewise, total IGF-I as well as alternatively spliced IGF-I mRNA (class 1 and class 2 transcripts) were measured. A decrease in total GHR mRNA at parturition (P < 0.01) was associated with a specific decrease in GHR 1A mRNA (P < 0.001). The amount of alternatively spliced GHR mRNA (including GHR 1B and GHR 1C mRNA) did not change at parturition (P > 0.10). Total liver IGF-I mRNA and blood IGF-I concentrations were also decreased at parturition (P < 0.05 and P < 0.01, respectively). However, a decrease in IGF-I mRNA was observed for both class 1 and class 2 IGF-I transcripts (P < 0.01 and P < 0.05, respectively). We conclude that the reduced amount of GHR mRNA during early lactation is caused by a specific down-regulation of GHR 1A mRNA that was associated with decreased liver IGF-I mRNA and decreased blood IGF-I concentrations. These data provide evidence for independent regulation of GHR mRNA by mechanisms that discriminate between GHR P1 (transcribes GHR 1A) and alternative promoters that transcribe constitutive GHR mRNA. PMID- 10465264 TI - Promoter-specific regulation of the brain-derived neurotropic factor gene by thyroid hormone in the developing rat cerebellum. AB - Thyroid hormone (TH) plays a critical role in normal cerebellar development. However, the molecular mechanisms of TH action in the developing cerebellum are not fully understood. This action could be exerted in part through brain-derived neurotropic factor (BDNF), as cerebellar BDNF messenger RNA (mRNA) expression is lower, and replacement of BDNF partially reverses the abnormal neurogenesis in the hypothyroid rat. The rat BDNF gene consists of four noncoding exons (exons I IV), each of which is linked to a different promoter, and a protein-coding exon (exon V). To study promoter-specific regulation of the BDNF gene by TH, ribonuclease protection assay of each exon mRNA was performed using total developing rat cerebellar RNA. During cerebellar development, all exon mRNAs were detected, but with different expression patterns; among noncoding exon mRNAs, exon II mRNA was the most abundant. Daily TH replacement induced a 3-fold increase in exon II mRNA on postnatal day (P) 15. On P30, exon II mRNA was still much greater in the TH-replaced animal. Exon I mRNA was detected on P2 and P7. However, in contrast to exon II mRNA, TH treatment suppressed the expression of exon I mRNA on P2. Exon III and IV mRNAs were not detected on P2 and P7, but small amounts were observed starting on P15 in TH-replaced animals. They were not detected by P30 in hypothyroid animals. In contrast, in the cerebral cortex, although all exons are differentially regulated during development, the expression of each mRNA was not significantly altered by TH. These results indicate that TH regulates BDNF gene expression in a promoter-, developmental stage-, and brain region-specific manner, which may play an important role in region- and stage-specific regulation of brain development by TH. PMID- 10465265 TI - Differential effects of insulin-like growth factor I (IGF-I) and IGF-binding protein-1 on protein metabolism in human skeletal muscle cells. AB - Insulin-like growth factor-binding protein-1 (BP-1) is a multifunctional protein that binds IGF-I in solution and integrins on the cell surface. BP-1 is overexpressed during catabolic illnesses, and the protein accumulates in skeletal muscle. To define a potential physiological role for BP-1 in regulating muscle protein balance, we have examined the effect of IGF-I and BP-1 on protein synthesis and degradation in human skeletal muscle cells. IGF-I-stimulated protein synthesis by 20%, and this was completely inhibited by either phosphorylated or nonphosphorylated BP-1. Half-maximal inhibition of protein synthesis occurred at a molar ratio of BP-1 to IGF-I of 1.5:1. BP-1 failed to form a complex with a truncated form of IGF-I (desIGF-I), and consequently, BP-1 failed to inhibit the ability of desIGF-I to stimulate protein synthesis. IGF-I and BP-1 dose-dependently inhibited protein degradation individually, and both BP 1 phosphovariants failed to block the ability of IGF-I to do the same. Blocking integrin receptor occupancy with the integrin antagonist echistatin blunted the ability of BP-1 to inhibit protein degradation, but had no significant effect on IGF-I-mediated changes in protein synthesis or degradation. The extracellular matrix protein vitronectin also inhibited protein degradation, but vitronectin receptor antibodies failed to block BP-1 action. In contrast, antibodies to the beta1 integrin subunit blocked BP-1-mediated inhibition of protein degradation. Rapamycin inhibited IGF-I-dependent protein synthesis, but not the ability of IGF I to inhibit proteolysis. In contrast, rapamycin completely blocked the ability of BP-1 to inhibit proteolysis. Our results demonstrate that BP-1 inhibits IGF-I mediated protein synthesis by binding to IGF-I. BP-1, acting independently of IGF I, inhibits protein degradation. The IGF-independent response occurs via beta1 integrin binding and stimulation of a rapamycin-sensitive signal transduction pathway. PMID- 10465266 TI - Glucosamine regulation of glucose metabolism in cultured human skeletal muscle cells: divergent effects on glucose transport/phosphorylation and glycogen synthase in non-diabetic and type 2 diabetic subjects. AB - Chronic exposure (48 h) to glucosamine resulted in a dose-dependent reduction of basal and insulin-stimulated glucose uptake activities in human skeletal muscle cell cultures from nondiabetic and type 2 diabetic subjects. Insulin responsiveness of uptake was also reduced. There was no change in total membrane expression of either GLUT1, GLUT3, or GLUT4 proteins. While glucosamine treatment had no significant effects on hexokinase activity measured in cell extracts, glucose phosphorylation in intact cells was impaired after treatment. Under conditions where glucose transport and phosphorylation were down regulated, the fractional velocity (FV) of glycogen synthase was increased by glucosamine treatment. Neither the total activity nor protein expression of glycogen synthase were influenced by glucosamine treatment. The stimulation of glycogen synthase by glucosamine was not due totally to soluble mediators. Reflective of the effects on transport/phosphorylation, total glycogen content and net glycogen synthesis were reduced after glucosamine treatment. These effects were similar in nondiabetic and type 2 cells. In summary: 1) Chronic treatment with glucosamine reduces glucose transport/phosphorylation and storage into glycogen in skeletal muscle cells in culture and impairs insulin responsiveness as well. 2) Down regulation of glucose transport/phosphorylation occurs at a posttranslational level of GLUTs. 3) Glycogen synthase activity increases with glucosamine treatment. 4) Nondiabetic and type 2 muscle cells display equal sensitivity and responsiveness to glucosamine. Increased exposure of skeletal muscle to glucosamine, a substrate/precursor of the hexosamine pathway, alters intracellular glucose metabolism at multiple sites and can contribute to insulin resistance in this tissue. PMID- 10465267 TI - Defense of adrenocorticosteroid receptor expression in rat hippocampus: effects of stress and strain. AB - Neuronal mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) proteins are glucocorticoid-activated transcription factors that bind identical DNA response elements yet transduce distinct physiological/transcriptional actions. The present study assessed regulation of adrenocorticosteroid receptor RNA and protein following intermittent stress exposure, using Sprague Dawley (S D) and stress-hyperresponsive Fischer 344 (F344) rat strains. The F344 (but not S D) strain showed enhanced acute stress responsivity and enhanced corticosterone secretion following prolonged stress. F344 rats also showed reduced responsiveness to a novel stressor after prolonged stress exposure, suggestive of enhanced glucocorticoid negative feed-back. Upon prolonged stress, F344 rats down regulated MR hnRNA in CA1, CA3, and dentate gyrus. Transcriptional changes were accompanied by decreased expression of the alpha 5' messenger RNA (mRNA) form, consistent with altered promoter utilization. In contrast, alpha 5' splice variant, full-length mRNA, and MR protein expression were not affected by stress in either strain, implying that transcriptional changes do not affect overall mRNA or protein expression. GR protein was increased in pyramidal and granule cell somata/nuclei of F344 rats despite lack of a change in mRNA expression. These data suggest that prolonged stress elicits restricted changes in MR and GR expression in the F344 strain only. Overall, stable expression of adrenocorticosteroid receptors is rigorously defended in hippocampal neurons, apparently through transcriptional and posttranscriptional mechanisms. PMID- 10465268 TI - The role of amino acids surrounding tyrosine 1062 in ret in specific binding of the shc phosphotyrosine-binding domain. AB - We investigated the role of the I-E-N-K-L (amino acids 1057-1061) sequence amino terminal to Tyr1062 in Ret for binding of the Shc phosphotyrosine-binding (PTB) domain. Substitution of Ser for Ile1057 (I1057S), Ala for Asn1059 (N1059A), or Pro for Leu1061 (L1061P) in this sequence significantly decreased the transforming activity of Ret with the multiple endocrine neoplasm type 2A (MEN2A) mutation as well as the binding affinity of Shc to it in vivo and in vitro, indicating that these three amino acids play a role in Shc binding. In addition, as the RET protooncogene is translated as three isoforms of 1114 amino acids (Ret 51), 1106 amino acids (Ret 43), and 1072 amino acids (Ret 9) that differ from one another in the sequence carboxyl-terminal to Tyr1062, we examined whether these sequence differences influence the binding affinity of Shc to Ret. As a result, we found that the transforming activity of Ret 43 isoform with the MEN2A mutation and the binding affinity of Shc to it were very low, although the consensus sequence for the binding of the Shc PTB domain is conserved in the Ret 43 isoform. This finding suggested that the sequence carboxyl-terminal to Tyr1062 in Ret could also influence the binding affinity to Shc. PMID- 10465269 TI - Preserved pulsatile insulin release from prediabetic mouse islets. AB - During the development of type I diabetes, the plasma insulin pattern changes. Because the islet secretory pattern has been implicated in this phenomenon, insulin release was measured from female nonobese diabetic (NOD) mouse islets isolated at different ages. Islets from 5-week-old mice were used as controls because they had no infiltrating mononuclear cells and insulin release rose almost 9-fold with maintained oscillatory frequency when the glucose concentration was raised from 3 to 11 mM. Islets isolated from 13- and 25-week old mice were infiltrated with mononuclear cells. In these islets, increase in the glucose concentration from 3 to 11 mM only doubled insulin release. However, despite the cellular infiltration, insulin release was pulsatile. Islets from 13 week-old mice had reduced glucose oxidation rate. Culture of such islets for 7 days at 11.1 mM glucose causes a decrease in the number of mononuclear cells infiltrating the islets, which in the present study was accompanied by a normalization of both glucose oxidation and glucose-induced insulin release. In the presence of the mitochondrial substrate alpha-keto-isocaproate (5 mM) both control and infiltrated islets responded with pronounced insulin pulses with similar amplitudes. The results suggest that the deranged plasma insulin pattern observed during the development of type I diabetes may be related to decrease in the insulin pulse amplitude rather than loss of the pulsatile release from the islets. PMID- 10465270 TI - Glucose-induced preproinsulin gene expression is inhibited by the free fatty acid palmitate. AB - Prolonged exposure to elevated FFA levels has been shown to induce peripheral insulin resistance and to alter the beta-cell secretory response to glucose. To investigate the effects of FFAs on preproinsulin gene expression, we measured insulin release, cell content, and messenger RNA (mRNA) levels in rat islets after a 24-h exposure to 1 mM palmitate. Insulin release increased at all glucose concentrations studied; in contrast, preproinsulin mRNA levels were specifically reduced by palmitate at high glucose with a decrease in insulin stores, suggesting that palmitate inhibits the glucose-stimulated increase in preproinsulin gene expression. The mechanisms by which palmitate affects preproinsulin gene expression implicate both preproinsulin mRNA stability and transcription, as suggested by an actinomycin D decay assay, quantification of primary preproinsulin transcripts, and transient transfection experiments in Min6 cells. Metabolism of palmitate is not required to obtain these effects, inasmuch as they can be reproduced by 2-bromopalmitate. However, oleate and linoleate did not significantly influence preproinsulin mRNA levels. We conclude that insulin release and preproinsulin gene expression are not coordinately regulated by palmitate and that chronically elevated FFA levels may interfere with beta-cell function and be implicated in the development of noninsulin-dependent diabetes. PMID- 10465271 TI - Starvation: early signals, sensors, and sequelae. AB - To identify the sequences of changes in putative signals, reception of these and responses to starvation, we sampled fed and starved rats at 2- to 6-h intervals after removal of food 2 h before dark. Metabolites, hormones, hypothalamic neuropeptide expression, fat depots, and leptin expression were measured. At 2 h, insulin decreased, and FFA and corticosterone (B) increased; by 4 h, leptin and glucose levels decreased. Neuropeptide Y messenger RNA (mRNA) increased 6 h after food removal and thereafter. Adrenal and plasma B did not follow ACTH and were elevated throughout, with a nadir at the dark-light transition. Leptin correlated inversely with adrenal B. Fat stores decreased during the last 12 h. Leptin mRNA in perirenal and sc fat peaked during the dark period, resembling plasma leptin in fed rats. We conclude that 1) within the first 4 h, hormonal and metabolic signals relay starvation-induced information to the hypothalamus; 2) hypothalamic neuropeptide synthesis responds rapidly to the altered metabolic signals; 3) catabolic activity quickly predominates, reinforced by elevated B, not driven by ACTH, but possibly to a minor extent by leptin, and more by adrenal neural activity; and 4) leptin secretion decreases before leptin mRNA or fat depot weight, showing synthesis-independent regulation. PMID- 10465272 TI - Growth hormone-independent cardioprotective effects of hexarelin in the rat. AB - We previously reported that induction of selective GH deficiency in the rat exacerbates cardiac dysfunction induced by experimental ischemia and reperfusion performed on the explanted heart. In the same model, short-term treatment with hexarelin, a GH-releasing peptide, reverted this effect, as did GH. To ascertain whether hexarelin had non-GH-mediated protective effects on the heart, we compared hexarelin and GH treatment in hypophysectomized rats. Hexarelin (80 microg/kg sc), given for 7 days, prevented exacerbation of the ischemia reperfusion damage induced by hypophysectomy. We also demonstrate that hexarelin prevents increases in left ventricular end diastolic pressure, coronary perfusion pressure, reactivity of the coronary vasculature to angiotensin II, and release of creatine kinase in the heart perfusate. Moreover, hexarelin prevents the fall in prostacyclin release and enhances recovery of contractility. Treatment with GH (400 microg/kg sc) produced similar results, whereas administration of EP 51389 (80 microg/kg sc), another GH-releasing peptide that does not bind to the heart, was ineffective. In conclusion, we demonstrate that hexarelin prevents cardiac damage after ischemia-reperfusion, and that its action is not mediated by GH but likely occurs through activation of specific cardiac receptors. PMID- 10465273 TI - Transcriptional activation of the decidual/trophoblast prolactin-related protein gene. AB - The decidual/trophoblast PRL-related protein (d/tPRP) is dually expressed by decidual and trophoblast cells during pregnancy. We have characterized the proximal d/tPRP promoter responsible for directing d/tPRP expression in decidual and trophoblast cells. We have demonstrated that the proximal 93 bp of d/tPRP 5' flanking DNA are sufficient to direct luciferase gene expression in primary decidual and Rcho-1 trophoblast cells, but not in fibroblast, undifferentiated uterine stromal cells or trophoblast cells of a labyrinthine lineage. The 93-bp d/tPRP promoter was also sufficient to direct differentiation-dependent expression in trophoblast giant cells. Mutational analysis demonstrated the differential importance of activating protein-1 and Ets regulatory elements (located within the proximal 93 bp of d/tPRP 5'-flanking DNA) for activation of the d/tPRP promoter in decidual vs. trophoblast cells. Disruption of the activating protein-1 regulatory element inhibited d/tPRP promoter activity by more than 95% in decidual cells, and approximately 80% trophoblast cells. Disruption of the Ets regulatory element reduced d/tPRP promoter activity by approximately 50% in decidual cells, while inactivating the d/tPRP promoter in trophoblast cells. Protein interactions with the trophoblast Ets regulatory element were shown to be cell type specific and to change during trophoblast giant cell formation. In conclusion, a 93-bp region of the d/tPRP promoter is shown to contain regulatory elements sufficient for gene activation in decidual and trophoblast cells. PMID- 10465274 TI - The role of cell surface attachment and proteolysis in the insulin-like growth factor (IGF)-independent effects of IGF-binding protein-3 on apoptosis in breast epithelial cells. AB - We have recently reported that insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) can significantly increase ceramide-induced apoptosis in an Hs578T breast carcinoma cell line in an IGF-independent manner. It was observed in that study that IGFBP-3 added to the cultures was proteolytically modified, generating a specific pattern of fragmentation. We have also previously reported that almost all of the IGFBP-3 outside the circulation in extravascular fluids is in a fragmented form, apparently due to the activity of a cation-dependent serine protease. The aim of this study was to investigate the role of proteolysis in the IGFBP-3 enhancement of C2-induced apoptosis. In this study we confirmed that preincubation of Hs578T cells with IGFBP-3 enhances the apoptotic effect of the ceramide analog C2. The presence of IGF-I completely inhibited the enhancement effect, apparently by inhibiting cell surface association and proteolytic modification. The presence of a serine protease inhibitor [4-(2 aminoethyl)benesulfonyl fluoride] completely inhibited the enhancement effect of IGFBP-3, and Western immunoblotting of conditioned medium and cell surface associated IGFBP-3 revealed that proteolytic fragmentation of the IGFBP-3 was reduced. In addition, fragments from the incubation of IGFBP-3 with plasmin were able to enhance the susceptibility of Hs578T cells to C2. The effect of these fragments could, however, also be reduced by 4-(2-aminoethyl)benesulfonyl fluoride despite the fact that IGFBP-3 was already fragmented. This suggests additional roles for serine proteases in the IGFBP-3 effect on C2-induced apoptosis in addition to the cleavage of the binding protein. PMID- 10465275 TI - Evidence that the inhibition of luteinizing hormone secretion exerted by central administration of neuropeptide Y (NPY) in the rat is predominantly mediated by the NPY-Y5 receptor subtype. AB - A number of studies have indicated that neuropeptide Y (NPY) is a central regulator of the gonadotropic axis, and the Y1 receptor was initially suggested to be implicated. As at least five different NPY receptor subtypes have now been characterized, the aim of the present study was to reinvestigate the pharmacological profile of the receptor(s) mediating the inhibitory action of NPY on LH secretion by using a panel of NPY analogs with different selectivity toward the five NPY receptor subtypes. When given intracerebroventricularly (icv) to castrated rats, a bolus injection of native NPY (0.7-2.3 nmol) dose-dependently decreased plasma LH. Peptide YY (PYY; 2.3 nmol) was as potent as NPY, suggesting that the Y3 receptor is not implicated. Confirming previous data, the mixed Y1, Y4, and Y5 agonist [Leu31,Pro34]NPY (0.7-2.3 nmol) inhibited LH release with potency and efficacy equal to those of NPY. Neither the selective Y2 agonist C2 NPY (2.3 nmol) nor the selective Y4 agonist rat pancreatic polypeptide affected plasma LH, excluding Y2 and Y4 subtypes for the action of NPY on LH secretion. The mixed Y4-Y5 agonist human pancreatic polypeptide (0.7-7 nmol) as well as the mixed Y2-Y5 agonist PYY3-36 (0.7-7 nmol) that displayed very low affinity for the Y1 receptor, thus practically representing selective Y5 agonists in this system, decreased plasma LH with potency and efficacy similar to those of NPY, indicating that the Y5 receptor is mainly involved in this inhibitory action of NPY on LH secretion. [D-Trp32]NPY, a selective, but weak, Y5 agonist, also inhibited plasma LH at a dose of 7 nmol. Furthermore, the inhibitory action of NPY (0.7 nmol) on LH secretion could be fully prevented, in a dose-dependent manner (6-100 microg, icv), by a nonpeptidic Y5 receptor antagonist. This antagonist (60 microg, icv) also inhibited the stimulatory action of NPY (0.7 nmol) on food intake. The selectivity of PYY3-36, human PP, [D-Trp32]NPY, and the Y5 antagonist for the Y5 receptor subtype was further confirmed by their ability to inhibit the specific [125I][Leu31,Pro34]PYY binding to rat brain membrane homogenates in the presence of the Y1 receptor antagonist BIBP3226, a binding assay system that was described as being highly specific for Y5-like receptors. With the exception of [D Trp32]NPY, all analogs able to inhibit LH secretion were also able to stimulate food intake. Taken together, these results indicate that the Y5 receptor is involved in the negative control by NPY of the gonadotropic axis. PMID- 10465276 TI - Regulation of urokinase production by androgens in human prostate cancer cells: effect on tumor growth and metastases in vivo. AB - During the complex multistep process of tumor progression, prostate cancer is initiated as an androgen-sensitive, nonmetastatic cancer, followed by a gradual transition into a highly metastatic and androgen-insensitive variety that lacks the expression of functional androgen receptors (AR). Urokinase (uPA), a member of the serine protease family, has been implicated in the progression of various human malignancies, including prostate cancer. Although uPA production is regulated by various growth factors and cytokines, the role of sex steroids (androgens) in regulating uPA gene expression in prostate cancer is poorly understood. In the current study, we have examined the role of androgens in regulating uPA production and the invasive capacity of the androgen insensitive PC-3 cells transfected with the full-length human AR complementary DNA (PC-3T). Restoration of androgen responsiveness in PC-3T cells caused a marked decrease in cell doubling time. Treatment of PC-3T cells with dihydroxytestosterone (DHT) caused a dose-dependent decrease in uPA messenger RNA and protein production, resulting in their decreased ability to invade through the Matrigel. Nuclear runoff assays revealed that these effects were attributable to the ability of DHT to inhibit uPA gene transcription. AR antagonist flutamide (Flu) reversed the effect of DHT on proliferation and invasion of PC-3T cells. Both control (PC-3) and experimental (PC-3T) cells were injected into the right flank of male BALB/c nu/nu mice. Control animals developed palpable tumors and microscopic tumor metastases at lymph nodes, lungs, and liver at 6-week posttumor cell inoculation. In contrast to this, because of androgen sensitivity of PC-3T cells, palpable tumors were observed only at week 12, with occasional tumor metastases in lungs. Furthermore, inoculation of PC-3T cells into surgically castrated host animals resulted in the development of tumors at a much earlier time (week 10) and a high incidence of metastases, compared with regular animals receiving PC-3T cells. Collectively, these results demonstrate the ability of androgen to regulate uPA production, which may directly effect prostate cancer growth, invasion, and metastasis in vitro and in vivo. PMID- 10465277 TI - Stimulatory actions of insulin-like growth factor-I and transforming growth factor-alpha on intestinal neurotensin and peptide YY. AB - Proliferation of the gastrointestinal mucosa is stimulated by the growth factors, insulin-like growth factor-I (IGF-I) and transforming growth factor-alpha (TGF alpha), or the closely related epidermal growth factor (EGF), as well as the gastrointestinal hormones, gastrin, neurotensin (NT), and peptide YY (PYY). The stimulatory actions of these growth factors or gastrointestinal hormones on the gastrointestinal mucosa may be direct or mediated in part by gastrointestinal peptides or the growth factors, respectively. The purpose of these studies therefore was to examine the effects of IGF-I and TGF-alpha on stomach gastrin and intestinal NT and PYY gene expression [i.e. messenger RNA (mRNA), peptide levels] and secretion. Mice were given recombinant human IGF-I (3, 6 mg/kg BW/day x 14 days). Transgenic mice with the rat TGF-alpha gene linked to a metallothionein promoter were used as a model of chronic TGF-alpha excess. IGF-I and TGF-alpha did not affect gastrin gene expression. Steady-state intestinal NT and PYY mRNA and peptide levels were elevated in a dose-related manner by IGF. TGF-alpha also increased intestinal expression of NT and PYY peptide, but not mRNA levels. Basal serum levels of PYY were elevated by IGF-I and TGF-alpha. IGF I and TGF-alpha did not increase intestinal chromogranin A (CGA) gene expression, a marker of endocrine cells, or the density of PYY-containing cells in the colon, indicating that the elevations in intestinal gut peptide gene expression by IGF-I and TGF-alpha are not due simply to an increased number of enteroendocrine cells. IV infusion of EGF also stimulated release of PYY in the dog. Together, these findings indicate that IGF-I and TGF-alpha may cause secretion of gut hormones and exert a major upregulatory influence on the regulation of intestinal peptide hormone homeostasis. PMID- 10465278 TI - Discovery and characterization of endometrial epithelial messenger ribonucleic acids using the ovine uterine gland knockout model. AB - Prolonged exposure of the developing neonatal ovine uterus to a progestin from birth prevents uterine gland development and creates an adult endometrial phenotype characterized by the absence of glandular epithelium, the uterine gland knockout (UGKO) phenotype. This study used endometrium from normal and UGKO sheep to identify messenger RNAs (mRNAs) expressed differentially in the endometrial epithelium using the molecular techniques of mRNA differential display PCR (DD PCR) and suppression subtractive complementary DNA (cDNA) hybridization (SSH). Sequence analyses of DD- and SSH-identified and cloned cDNAs indicated similarity of some to known mRNAs, including beta-lactoglobulin, alkaline phosphatase, type B and D endogenous sheep retroviruses, gp330/megalin, matrix Gla protein, and others. Other cDNAs were not similar to any known sequences and are considered novel, although some of these match human expressed sequence tags. In situ hybridization analyses of uteri from cyclic and pregnant ewes indicated that all DD-PCR- and SSH-identified mRNAs were expressed in either the endometrial lumenal and/or glandular epithelium, although some were also expressed in other uterine cell types. Northern and in situ hybridization analyses revealed that patterns of mRNA expression for most clones were affected by the day of the estrous cycle and pregnancy in a manner consistent with regulation by progesterone. Studies demonstrate the utility of the ovine UGKO model as a tool with which to identify known and novel uterine epithelial-specific genes. Cloned cDNAs identified here are expressed sequence tags useful for comparative and physical genetic mapping and may be used to reveal new factors and pathways regulating endometrial function. PMID- 10465279 TI - Hypothalamic obesity: multiple routes mediated by loss of function in medial cell groups. AB - Cell groups of the medial hypothalamus are key to the regulation of energy balance. Functional disruption by colchicine injected in the hypothalamic arcuate (ARC), paraventricular (PVN), and ventromedial (VMN) cell groups produced increased food intake and obesity; disruption of the dorsomedial nuclei (DMN) produced decreased food intake. Colchicine in ARC or PVN increased food intake during both light and dark periods and increased cumulative food intake. By contrast, colchicine in VMN increased food intake only during the light, and cumulative food intake was not increased. Both leptin and insulin were elevated in the obese rats. Compared with sham, the slope of regression of leptin on insulin was increased by disruption of PVN and DMN but was not altered by disruption of VMN. ARC disruption abolished the relationship between leptin and insulin. Colchicine injected in the DMN did not cause obesity but altered feeding and the normal relationship between leptin, fat, and insulin, suggesting that blockade of signals, for example, from the lateral hypothalamus to DMN may disinhibit the normal medial hypothalamic drive to decrease energy stores. Changes in caloric efficiency with time after colchicine injections suggest that rats with both ARC and PVN disruption respond to signals of obesity, whereas rats with VMN disruption do not. These studies distinguish among functions in the four medial hypothalamic nuclei and suggest that interactions among them normally serve to regulate energy balance through alterations in food acquisition and storage. PMID- 10465280 TI - Growth hormone and insulin-like growth factors have different effects on sebaceous cell growth and differentiation. AB - Several observations suggest that GH stimulates sebaceous gland growth and development. Therefore, we studied the effects of GH and insulin-like growth factors (IGFs), alone and with androgen, on sebaceous epithelial cell (sebocyte) growth and differentiation in vitro. The rat preputial cell culture model system was used to judge differentiation (induction of lipid-forming colonies, LFCs) and DNA synthesis. GH increased sebocyte differentiation. At a dose of 10(-8) M in the presence of micromolar insulin, GH was 3.8 times more potent than IGF-I (38.1+/-4.2%, SEM, vs. 10+/-1.5% LFCs) and 6 times more potent than IGF-II (6+/ 0.5% LFCs). IGF-I 10(-8) M alone stimulated a similar amount of differentiation as insulin 10(-6) M, although it was less effective than insulin in augmenting the effect of GH on differentiation. GH had no effect on sebocyte uptake of 3H thymidine at doses up to 10(-6) M. On the other hand, IGF-I was the most potent stimulus of DNA synthesis (168% of control; P < 0.001 vs. all others). IGF-II 10( 8) M stimulated 3H-thymidine incorporation similarly to insulin 10(-6) M. In the presence of insulin, dihydrotestosterone (DHT) 10(-6) M induced 31.4+/-1.7% LFCs, and there was a tendency of DHT and GH to interact in promoting differentiation. When insulin was omitted from the system, differentiation was decreased overall, but GH +/- DHT slightly improved differentiation. The IGFs had no effect on the response to DHT. DHT decreased DNA synthesis by 40%, an effect unaltered by GH or IGFs. These results suggest that GH and IGFs have different functions in sebaceous cell growth and differentiation: GH stimulated differentiation beyond that found with IGFs or insulin, yet had no effect on DNA synthesis, a parameter stimulated most potently by IGF-I. While GH augmented the effect of DHT on differentiation, the IGFs had no effect on the response of DHT. These data indicate that GH may in part act directly on sebocytes rather than indirectly through IGF production. These data are consistent with the concept that increases in GH and IGF production contribute in complementary ways to the increase in sebum production during puberty and in acromegaly. PMID- 10465281 TI - Transcriptional regulation of corticotropin-releasing hormone-binding protein gene expression in astrocyte cultures. AB - The molecular mechanisms involved in regulation of CRH-binding protein (CRH-BP) gene expression were examined using primary rat astrocyte cultures. The cells were treated with various regulators, and CRH-BP messenger RNA (mRNA) levels were determined using ribonuclease protection assays. Forskolin (Fsk, 10 microM) or 12 O-tetradecanoyl-phorbol 13-acetate (TPA, 100 nM) increases CRH-BP mRNA levels up to 30 times control level, and together they act synergistically to increase CRH BP gene expression up to 100 times control levels. CRH can also positively regulate CRH-BP gene expression to 6.1 times control levels. All of these increases in steady-state CRH-BP mRNA levels can be repressed by dexamethasone, a synthetic glucocorticoid. To determine whether these changes in steady-state CRH BP mRNA levels are caused by altered transcription or RNA stability, heteronuclear (hn) CRH-BP species were examined using ribonuclease protection assays. CRH-BP hnRNA transcripts can be detected transiently after the addition of Fsk or TPA, and dexamethasone can repress Fsk- or TPA-induced CRH-BP hnRNA levels in this assay. These results demonstrate that CRH, glucocorticoids, and the protein kinase A and protein kinase C signaling pathways are involved in regulation of CRH-BP gene expression in astrocyte cultures, and that this regulation is caused, at least in part, by altered transcription of the gene. PMID- 10465282 TI - Frog chromogranin A messenger ribonucleic acid encodes three highly conserved peptides. Coordinate regulation of proopiomelanocortin and chromogranin A gene expression in the pars intermedia of the pituitary during background color adaptation. AB - Chromogranin A (CgA) is a neuroendocrine secretory protein that is widely used as a marker for endocrine neoplasms but whose function is not completely understood. In mammals, it is thought that CgA is a precursor for biologically active peptides. Here, we describe the cloning of a complementary DNA encoding CgA from a nonmammalian vertebrate, the frog Rana ridibunda. Sequence analysis revealed that frog CgA exhibits only 40-44% amino acid sequence similarity with its mammalian homologues. The amino acid identity is confined to three regions (70 80% identity) of the protein that are flanked by conserved pairs of basic amino acid residues, suggesting that proteolytic processing at these cleavage sites may give rise to three biologically active peptides whose sequences have been highly preserved during evolution. Tissue distribution analysis by Northern blot and in situ hybridization revealed the widespread expression of frog CgA messenger RNA in the brain and in endocrine tissues, the highest concentration occurring in the distal lobe of the pituitary. Adaptation of frog skin color to a dark background caused a concomitant increase in CgA and POMC messenger RNA levels in the intermediate lobe of the pituitary. Taken together, these data indicate that CgA may function as a precursor to three highly conserved peptides that may exert regulatory functions in the neuroendocrine system. PMID- 10465283 TI - Triiodothyronine down-regulates thyrotropin-releasing hormone (TRH) synthesis and decreases pTRH-(160-169) and insulin releases from fetal rat islets in culture. AB - TRH is localized with insulin in beta-cells. It is synthesized as a prohormone containing five copies of TRH and seven cryptic peptides, including pro (p)-TRH (160-169). Thyroid hormone regulates the expression of several genes encoding peptide hormones. We found that circulating T3 concentrations were inversely correlated with TRH levels in two physiopathological situations. There are low concentrations of circulating thyroid hormone and very high concentrations of TRH and pTRH-(160-169) during development, and experimental hypothyroidism results in higher concentrations of prepro (pp)-TRH messenger RNA (mRNA) and TRH content in the adult rat pancreas than are present in the euthyroid pancreas. The present study was carried out to investigate the interaction between T3 and pancreatic TRH during the functional maturation of islets in culture and to validate the data obtained in vivo. T3 decreases ppTRH mRNA in islets in a dose-dependent manner. The primary impact of T3 on islet function may be mediated by ppTRH mRNA, as short term T3 treatment had no effect. Long term T3 treatment reduced the islet TRH content and the amounts of pTRH-(160-169) and insulin released. This secretory pattern and coordinated regulation of pTRH-(160-169) and insulin suggests that pTRH-(160-169) plays a specific role in the regulation of insulin secretion. PMID- 10465284 TI - Transcriptional repression of the rat osteocalcin gene: role of two intronic CCTCCT motifs. AB - The regulation of osteocalcin gene transcription is complex, involving multiple positive and negative regulators. Previous studies have demonstrated that an intronic sequence, TTTCTTT (+118 to +124) is capable of mediating transcriptional repression of osteocalcin-CAT fusion genes in cells of the osteoblast lineage, by interacting with a specific nuclear protein. Further analyses of intronic sequences have identified a second silencer motif in this region. Two copies of a CCTCCT motif are present within the first intron of the rat osteocalcin gene (+106 to +111 and +135 to +140) and are capable of mediating transcriptional repression of osteocalcin-CAT fusion genes in rat osteosarcoma cells. Transient gene expression assays of wild-type and mutant osteocalcin-CAT fusion genes into ROS 17/2.8 cells demonstrate that mutagenesis of either of these CCTCCT motifs in isolation results in a 1.6-fold increase in CAT activity relative to the parent fusion gene. Moreover, a 5-fold increase in reporter gene activity is observed when both motifs are mutated together. These sequences are also capable of suppressing osteocalcin promoter activity when placed upstream to the osteocalcin promoter. Gel retardation and southwestern analyses demonstrate that the CCTCCT motifs interact with specific proteins present in nuclear extracts from ROS 17/2.8 and UMR 106 osteosarcoma cells but not COS-7 kidney cells. Mutations that abolish suppressor function of this motif markedly impair interactions with this specific nuclear protein. These data demonstrate that at least two different silencer motifs (TTTCTTT and CCTCCT) in the first intron of the rat osteocalcin gene contribute to its transcriptional repression. PMID- 10465285 TI - Proteolysis of human prolactin: resistance to cathepsin D and formation of a nonangiostatic, C-terminal 16K fragment by thrombin. AB - The N-terminal 16K fragments of rat and human PRLs possess angiostatic activity. 16K PRL has also been detected in vivo in both humans and rats. Based on an in vitro study, cathepsin D, an acid protease, has been implicated in the generation of rat 16K PRL. However, the proteolytic cleavage of human PRL has not been demonstrated. Our objective was to identify an enzyme that is capable of forming an angiostatic human 16K PRL. To confirm the angiostatic action of rat 16K PRL, the fragment was generated by incubating 23K PRL with rat mammary microsomal fraction at pH 3.2. Upon incubation with human umbilical vein endothelial cells (HUVEC), rat 16K PRL, but not 23K PRL, inhibited basal- and basic fibroblast growth factor-stimulated cell proliferation. Intact rat and human PRLs were then incubated with cathepsin D or acidified microsomal pellets of MCF-7 human breast cancer cells. Analysis by SDS-PAGE showed cleavage of rat, but not human, PRL. Next, hormones were incubated with thrombin at pH 7.4. As shown by SDS-PAGE, digestion of both human and rat PRL by thrombin resulted in the formation of 16K fragments. PRL contained within human amniotic fluid was also cleaved by thrombin. Enzyme specificity was supported by prevention of cleavage by the thrombin inhibitor hirudin. When tested with HUVEC, the human 16K PRL was devoid of angiostatic activity. The activity of this fragment in the Nb2 lymphoma bioassay was 10- to 15-fold lower than that of 23K PRL. Mass spectrometry revealed that the fragment has a mass of 16,878.30+/-15.8 Daltons. Subsequent N terminal sequencing showed that the thrombin cleavage occurred between amino acid residues 53 (Lys) and 54 (Ala), resulting in the formation of a C-terminal, not an N-terminal, 16K fragment. We conclude that, unlike rat PRL, human PRL is resistant to cleavage by cathepsin D. Thrombin at a physiological pH can generate a C-terminal 16K fragment of human PRL that is not angiostatic and retains little mitogenic activity. We suggest that the precise nature of endogenous 16K PRL fragments that are present in human tissues and body fluids should be carefully examined. PMID- 10465286 TI - Dual regulation of promoter II- and promoter 1f-derived cytochrome P450 aromatase transcripts in equine granulosa cells during human chorionic gonadotropin-induced ovulation: a novel model for the study of aromatase promoter switching. AB - Estradiol biosynthesis is a key biochemical trait of developing follicles. To study its regulation in equine follicles, the objectives of this study were to clone and determine the structure of equine cytochrome P450 aromatase (P450AROM), and characterize the regulation of P450AROM and P450 17alpha-hydroxylase/C17-20 lyase (P45017alpha) messenger RNAs (mRNAs) in vivo in equine preovulatory follicles isolated during hCG-induced ovulation. Two distinct P450AROM complementary DNAs (cDNAs) were isolated from an equine preovulatory follicle cDNA library. One clone was 2682 bp in length and included 115 bp of 5' untranslated region (UTR), 1509 bp of open reading frame encoding a well conserved 503-amino acid protein, and 1058 bp of 3'-UTR. Its 5'-most region represented the equine homolog of exon 1f, previously designated brain specific. The other cDNA clone encoded a truncated protein and contained a distinct 5'-UTR characteristic of transcripts derived from promoter II, previously identified as the predominant ovarian mRNA. Northern blot analyses were performed using preovulatory follicles obtained during estrus between 0-39 h after the administration of hCG and with corpora lutea isolated on day 8 of the estrous cycle (day 0 = day of ovulation). The results showed a biphasic regulation of P450AROM mRNA expression: levels were highest in follicles at 0 h post-hCG, decreased significantly during the ovulatory process at 12 and 24 h (P < 0.05), and increased again between 30-39 h post-hCG and in corpora lutea. When oligonucleotides specific for P450AROM mRNA variants were used as probes, a novel switching phenomenon was observed. Promoter II-derived transcripts accounted for the message present in follicles at 0 h post-hCG and in corpora lutea, whereas promoter 1f-derived mRNA was expressed exclusively during the ovulatory process (30-39 h post-hCG). Levels of P45017alpha mRNA were high in follicles at 0 h, but significantly decreased after hCG treatment (P < 0.05), with lowest levels in follicles at 36 and 39 h post-hCG and in corpora lutea. Northern blots performed on isolated cellular preparations revealed that P450AROM and P45017alpha transcripts were localized exclusively in granulosa cells and theca interna, respectively. Equine aromatase promoters II and 1f were cloned from a genomic library, and putative transcription start sites were characterized by primer extension assays. Sequence analyses identified distinct potential regulatory elements in each promoter. Thus, this study identifies a novel aromatase promoter switching phenomenon in equine granulosa cells during follicular luteinization and provides a new model in which aromatase promoter switching is induced in vivo. PMID- 10465287 TI - Hexachlorobenzene, a dioxin-type compound, increases malic enzyme gene transcription through a mechanism involving the thyroid hormone response element. AB - Hexachlorobenzene (HCB) is a dioxin-type chemical that acts mainly through the aryl hydrocarbon receptor. Chronic exposure of rats to HCB increases the activity of malic enzyme (ME). In this report, we show that this increase is correlated with an induction of ME messenger RNA (mRNA) levels, with the maximal HCB effect achieved after 9 days of intoxication. This effect is specific for ME, as other liver enzymes, such as glyceraldehyde-3-phosphate dehydrogenase, phosphoenol pyruvate carboxykinase, and mitochondrial alpha-glycerol-3-phosphate dehydrogenase, are not affected by HCB. The induction of ME mRNA levels is accompanied by an increase in ME promoter activity, as demonstrated by transient transfection experiments performed in rat hepatoma H35 cells. In an attempt to identify the cis-regulatory elements responsible for the HCB effect, different promoter deletions and mutations were used. The results obtained localize the responsive region between positions -315 and -177. This region does not contain either consensus xenobiotic response or activating protein-1 elements, the two main mediators of dioxin compounds described to date. In contrast, a thyroid hormone response element (TRE) is located between -281 to -261. Deletions and mutations of the TRE element do not respond to HCB, demonstrating that this element mediates the response of this dioxin-type compound. As ME gene expression is regulated mainly by thyroid hormones, we next investigated the role of T3 receptor (T3R) in the ME gene transcriptional induction mediated by HCB. Using Scatchard analysis, we show that neither T3R binding features for its ligand nor alpha1 or beta1T3R mRNA levels are changed with the toxic. In gel shift assays, however, we observed that protein/DNA complexes formed on TRE from the ME promoter were induced by HCB. Using an oligonucleotide with a mutation that eliminates the TRE function, we demonstrate a loss of the induced protein/DNA complexes. Together, these data suggest that the dioxin-type compound HCB increases ME gene transcription by modulating the levels of still unidentified nuclear proteins that bind to the TRE element of the ME promoter. PMID- 10465288 TI - The rat growth hormone-releasing hormone receptor gene: structure, regulation, and generation of receptor isoforms with different signaling properties. AB - The interaction of GHRH with membrane-bound receptors on somatotroph cells of the anterior pituitary is an important step in the regulation of GH synthesis and secretion. The identification of a G protein-coupled receptor for GHRH has made it possible to investigate the pathway by which GHRH regulates pituitary somatotroph cell function. To initiate an analysis of the mechanisms regulating expression and function of the GHRH receptor, the structure of the gene and its promoter region were analyzed. The coding sequence of the rat GHRH receptor gene is contained within 14 exons spanning approximately 15 kb of genomic DNA. Four transcription start sites are located within 286 bp upstream of the initiation codon. The 5' flanking region of the GHRH receptor gene acts as a functional promoter in rat pituitary tumor GH3 cells, and basal promoter activity is enhanced in GH3 and COS7 cells by cotransfection of an expression construct encoding the pituitary-specific transcription factor Pit-1. The rat GHRH receptor gene is subject to at least 1 alternative RNA processing event that generates 2 receptor isoforms differing by 41 amino acids within the third intracellular loop (IL) of the protein. The short isoform of the GHRH receptor is predominant in pituitary cells. The MtT/S pituitary tumor cell line was found to express the GHRH receptor, and different populations of these cells produce predominantly the long or short isoforms of the receptor messenger RNA, suggesting that the alternative splicing can be regulated. Functional analysis of the two GHRH receptor isoforms demonstrates that both bind GHRH, but only the short isoform signals through a cAMP-mediated pathway. Neither receptor isoform is able to stimulate calcium mobilization from internal stores after GHRH treatment. Our findings indicate that the pituitary-specific transcription factor Pit-1 is involved in the somatotroph-specific expression of the GHRH receptor gene and that functionally distinct receptor proteins are generated by an alternative RNA processing mechanism. PMID- 10465289 TI - Expression and regulation of human sex hormone-binding globulin transgenes in mice during development. AB - Human sex hormone-binding globulin (SHBG) is produced by hepatocytes and transports sex steroids in the blood. The rat gene encoding SHBG is expressed transiently in the liver during fetal life, but it is not expressed in the liver postnatally, and the small amounts of SHBG in rat blood are derived from gonadal sources. To study the biosynthesis and function of human SHBG in an in vivo context, we have produced several lines of transgenic mice that contain either 11 kb (shbg11) or 4.3 kb (shbg4) portions of the human shbg locus. The expression and regulation of these transgenes have now been studied during fetal and postnatal development. In situ hybridization of an shbg11 transgenic mouse fetus at 17.5 days postcoitus located human shbg transcripts only in duodenal epithelial cells and hepatocytes. Temporal differences in the hepatic expression of mouse shbg and human shbg transgenes during late fetal development were reflected in corresponding differences in mouse and human SHBG levels in fetal and neonatal mouse blood. Serum concentrations of human SHBG increased during the first weeks of life regardless of gender until about 20 days of age in shbg11 mice, but after this time they continued to increase only in the males. This sexual dimorphism was reflected in corresponding differences in human SHBG messenger RNA (mRNA) abundance in the livers of these animals. However, it was not observed in shbg4 mice, in which hepatic production of plasma SHBG continued to increase after puberty regardless of gender. Serum testosterone and SHBG levels correlated in all sexually mature shbg transgenic mice. Human shbg transcripts were detectable only in testes of shbg11 mice and increased progressively in abundance from 10 days of age until the animal reached sexual maturity at 30 days of age, with appreciable increases occurring well before any changes in serum testosterone concentration. In the kidney, SHBG mRNA levels accumulated earlier in shbg11 than in shbg4 mice, and the expression of both types of transgenes was sexually dimorphic, with much higher SHBG mRNA levels in the kidneys of male mice. As increases in SHBG mRNA in the male kidneys coincided with increases in serum testosterone during sexual maturation, we reasoned that shbg transgene expression is androgen dependent in the kidney. This was confirmed by demonstrating that a decrease in SHBG mRNA abundance in male mouse kidneys after castration could be reversed by 5alpha-dihydrotestosterone treatment. Moreover, exogenous androgen increased human SHBG mRNA levels in the kidneys of female mice. In summary, comparisons of how different human shbg transgenes are expressed in vivo provides information about the positions of potential regulatory sequences that may control the hormonal regulation and tissue-specific expression of this gene during development. PMID- 10465290 TI - Insulin-like growth factor binding protein-4 proteolytic degradation in ovine preovulatory follicles: studies of underlying mechanisms. AB - The regulation of insulin-like growth factor binding protein (IGFBP)-4 proteolytic degradation by insulin-like growth factors (IGFs) has been largely studied in vitro, but not in vivo. The aim of this study was to investigate the involvement of IGFs, IGFBP-2, IGFBP-3, and IGFBP-3 proteolytic fragments in the regulation of IGFBP-4 proteolytic activity in ovine ovarian follicles. Follicular fluid from preovulatory follicles contains proteolytic activity degrading exogenous IGFBP-4. The addition of an excess of IGF-I enhanced IGFBP-4 proteolytic degradation, whereas addition of IGFBP-2 or -3 or monoclonal antibodies against IGF-I and -II dose dependently inhibited IGFBP-4 proteolytic degradation. IGF-I and IGF-II, but not LongR3-IGF-I, reversed this inhibition in a dose-dependent manner. C-terminal, but not N-terminal, proteolytic fragments derived from IGFBP-3 (aa 161-264), as well as heparin-binding domain-containing peptides derived from the C-terminal domain of IGFBP-3 and -5 also induced the inhibition of IGFBP-4 proteolytic degradation. Other heparin-binding domain containing peptides derived from the connective tissue growth factor (CTGF) and from proteins not related to IGFBP, heparan/heparin interacting protein (HIP) and vitronectin, but not from p36 subunit of annexin II tetramer, inhibited IGFBP-4 degradation. Furthermore, IGFBP-3, mutated on its heparin-binding domain, was not able to inhibit IGFBP-4 proteolytic degradation. So, in ovine preovulatory follicles, IGFBP-4 proteolytic degradation both 1) depends on IGFs, and 2) is inhibited by IGFBP-3 via its C-terminal heparin-binding domain as well as by heparin-binding domain containing peptides. These data suggest that in early atretic follicles, the increase in IGFBP-2 participates in the decrease in IGFBP 4 degradation. In late atretic follicles, the increase in the levels of C terminal IGFBP-3 proteolytic fragments, generated by IGFBP-3 degradation, as well as the increase in IGFBP-5 expression would strengthen the inhibition of IGFBP-4 degradation. This inhibition might be partly mediated by direct interaction of IGFBP-4 proteinase(s) and heparin-binding domain within the C-terminal region from IGFBP-3 and -5. PMID- 10465291 TI - beta1 to beta3 switch in control of cyclic adenosine monophosphate during brown adipocyte development explains distinct beta-adrenoceptor subtype mediation of proliferation and differentiation. AB - To explain the distinctive pharmacological profiles observed for adrenergic stimulation of cell proliferation (beta1) and cell differentiation (beta3), the adrenergic control of cAMP accumulation was investigated during brown adipocyte development. In preadipocytes, norepinephrine (NE) increased cAMP levels but the beta3-agonists BRL-37344 and CGP-12177 did not; in contrast, when the cells had differentiated into mature brown adipocytes, a large cAMP response to the beta3 agonists had emerged and was now double that to NE (although the affinity of NE had increased 10-fold). Beta1-messenger RNA (mRNA) levels were high in both pre- and mature brown adipocytes; beta3-mRNA did not appear until maturation but then abruptly. Although beta1-receptors remained detectable by [3H]CGP-12177 binding in the mature brown adipocytes, the cAMP response to NE (based on propranolol inhibitory potency) switched from beta1 to beta3. Even the established beta1 agonist dobutamine acted through beta3-receptors in the mature brown adipocytes. The increases in cAMP levels could adequately explain the increased cell proliferation in NE-stimulated preadipocytes and the NE-induced UCP1 gene expression in mature brown adipocytes. The distinctive adrenergic profiles for stimulation of proliferation and of differentiation were thus not due to the existence of additional pathways but to a switch in the type of beta-receptor mediating the NE response, coordinated with an alteration in the nuclear response to increased cAMP levels. The study implies that full recruitment of brown adipose tissue cannot be induced by exclusive beta3-stimulation. PMID- 10465292 TI - Inhibition of atrial wall stretch-induced cardiac hormone secretion by lavendustin A, a potent tyrosine kinase inhibitor. AB - The cellular processes linking mechanical wall stretch to atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) secretion from the heart are unclear. In the present study, a paced perfused rat heart preparation was used to study the signaling mechanisms of atrial wall stretch-induced secretion of ANP and BNP. Vehicle or drugs were infused into the perfusate for 40 min and right atrial wall stretch was superimposed for 10 min after 25-min drug infusions by elevating the level of the pulmonary artery cannula tip. Lavendustin A, a potent inhibitor of protein tyrosine kinases, at the concentrations of 0.5 and 1.3 microM decreased atrial wall stretch-induced ANP secretion (53% and 68%, respectively, P < 0.001) in the perfused rat heart preparation, whereas no difference in the hemodynamic variables (heart rate, contractile force and perfusion pressure) were noted between groups. Lavendustin A also completely abolished the wall stretch-induced secretion of BNP. Several other protein kinase inhibitors including staurosporine (protein kinase C inhibitor), ML-9 (myosin light chain kinase inhibitor), KN-62 (Ca2+/calmodulin-dependent protein kinase II inhibitor) and H-89 (protein kinase A inhibitor) had no significant effect on atrial wall stretch-stimulated ANP secretion. In a separate series of experiments, in which the right atria were stretched for 2 h, administration of lavendustin A (1 microM) but not staurosporine (30 nM) significantly decreased sustained wall stretch-induced ANP secretion. Okadaic acid, a potent protein phosphatase A2 (PPA2) and PP1 inhibitor, at the concentration of 100 nM had no effect on basal ANP secretion but significantly accelerated the ANP secretory response to atrial wall stretch (P < 0.05). In conclusion, the findings that inhibitors of protein tyrosine kinase and protein phosphatase selectively modulated atrial wall stretch-induced ANP secretion suggest a new mechanism involving endogenous protein tyrosine activity in the regulation of natriuretic peptide exocytosis from cardiac myocytes. PMID- 10465293 TI - Age-dependent inability of the endocrine pancreas to adapt to pregnancy: a long term consequence of perinatal malnutrition in the rat. AB - We have recently shown that maternal food restriction during late pregnancy decreased beta-cell mass in the offspring at birth. Prolonged maternal malnutrition until weaning led to irreversible decrease of beta-cell mass in the adult male progeny. During pregnancy, the maternal endocrine pancreas demonstrates an acute and reversible increase in beta-cell mass. The aim of this work was to investigate whether perinatal malnutrition could have long-lasting effects on glucose homeostasis and the adaptation of the endocrine pancreas to a subsequent pregnancy. This study was conducted on 4- and 8-month-old female rats malnourished during their perinatal life and on age-matched control animals. Oral glucose tolerance tests (OGTT), pancreatic insulin content, and islet mass quantitation after dithizone staining were performed on the same animals. Four month-old nonpregnant previously malnourished animals showed normal glucose tolerance but a significant decrease in insulin secretion during OGTT. These animals were, however, still able to adapt pancreatic insulin contents and doubled their islet mass in late gestation. At 8 months of age, insulin content before pregnancy was reduced to half that of controls. Moreover, it did not show the characteristic increase during gestation that could still be observed in pregnant control females. In those control animals, the islet mass increased regularly until late gestation (14.1+/-1.8 mg at day 20.5, vs. 9.8+/-1.2 mg, nonpregnant), whereas in previously malnourished animals the islet mass remained throughout pregnancy similar to the nonpregnant values (8.5+/-1.4 mg at day 20.5 vs. 8.9+/-3.6 mg, nonpregnant). In conclusion, early malnutrition has dramatic consequences on the capacity of the endocrine pancreas to meet the increased insulin demand during pregnancy and aging. PMID- 10465294 TI - Tumor necrosis factor-alpha and interferon-gamma suppress both gene expression and deoxyribonucleic acid-binding of TTF-2 in FRTL-5 cells. AB - Tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) are cytokines that can individually or additively suppress thyroid cell function and the expression of thyroid-specific genes, such as thyroglobulin (TG) and thyroperoxidase (TPO). Thyroid transcription factor-2 (TTF-2) is a DNA-binding protein that modulates the expression of TG and TPO genes. In the present study, we examine the effects of TNF-alpha and IFN-gamma on TTF-2 gene expression, as well as the DNA-binding activity of TTF-2. FRTL-5 cells were maintained in 5H medium containing 0.2% calf serum for 7 days, then incubated with TNF-alpha, IFN gamma, or TNF-alpha plus IFN-gamma. Total RNA was isolated and Northern blotted. TNF-alpha (50 ng/ml) only slightly suppressed (61+/-2% compared with control), whereas IFN-gamma (100 U/ml) modestly decreased TTF-2 messenger RNA (mRNA) levels (34+/-4%). TNF-alpha and IFN-gamma simultaneously caused a marked decrease in TTF 2 mRNA levels (13+/-2%). The suppressive effects of TNF-alpha and IFN-gamma on TTF-2 mRNA levels were concentration dependent and maximal at 50 ng/ml TNF-alpha with 100 U/ml IFN-gamma. The suppressive effect was also time dependent, reaching a maximum 12 h after exposure. Moreover, the suppressive effects of TNF-alpha and IFN-gamma upon rat TG and TTF-2 mRNA levels were similar. To test whether TNF alpha and IFN-gamma alter TTF-2-binding to DNA, we performed electrophoretic mobility shift assays using a TTF-2-binding element in the rat TG gene as a probe. Formation of the TTF-2/DNA complex was decreased by TNF-alpha and/or IFN gamma. Our results demonstrate that TNF-alpha and IFN-gamma additively reduce the gene expression and DNA-binding of TTF-2. These data suggest that TTF-2 is involved in the TNF-alpha and IFN-gamma-induced suppression of thyroid-specific gene expression. PMID- 10465295 TI - Thyroid hormone regulates the expression of laminin in the developing rat cerebellum. AB - In the rat cerebellum, migration of neurons from the external granular layer to the internal granular layer occurs postnatally and is dependent upon the presence of thyroid hormone. In hypothyroidism, many neurons fail to complete their migration and die. Key guidance signals to these migrating neurons are provided by laminin, an extracellular matrix protein that is fixed to the surface of astrocytes. Expression of laminin in the brain is developmentally timed to coincide with neuronal growth spurts. In this study, we examined the role of thyroid hormone on the expression and distribution of laminin in the rat cerebellum. We show that laminin content steadily increased 2- to 3-fold from birth to maximal levels on postnatal day 8-10 then steadily decreased to a plateau by postnatal day 12 in the euthyroid cerebellum. Immunoreactive laminin appeared in the molecular layer of the euthyroid cerebellum by postnatal day 4, reached maximal intensity by postnatal day 8-10, and was gone by postnatal day 14. In contrast, laminin content in the hypothyroid cerebellum remained unchanged from birth until postnatal day 10 and then increased to maximal levels over the next two days; maximal levels were approximately 35% less than those levels in the euthyroid cerebellum. Laminin staining did not appear in the molecular layer of the hypothyroid rat cerebellum until postnatal day 10, reached maximal intensity by postnatal day 15 and disappeared by postnatal day 18, despite the continued presence granular neurons in the external granular layer. These data indicate that the disruption of the timing of the appearance and regional distribution of laminin in the absence of thyroid hormone may play a major role in the profound derangement of neuronal migration observed in the cretinous brain. PMID- 10465296 TI - Roles of phosphatidylinositol 3-kinase and mitogen-activated protein kinase pathways in stimulation of vascular smooth muscle cell migration and deoxyriboncleic acid synthesis by insulin-like growth factor-I. AB - Insulin-like growth factor-I (IGF-I) is a potent stimulator of vascular smooth muscle cell (SMC) migration, a process that contributes to the accumulation of SMC within atherosclerotic lesions. Our previous studies have shown that IGF-I increases the affinity of the alphaVbeta3 integrin toward ligands and that occupancy of this integrin is indispensable for IGF-I to stimulate cell migration. In this study, the role of phosphatidylinositol 3-kinase (PI 3-kinase) and mitogen-activated protein kinase (MAPK) pathways in IGF-I induced cell motility and integrin activation was studied using porcine aortic smooth muscle cells (pSMC). Two structurally different inhibitors of PI 3-kinase decreased IGF I-stimulated pSMC migration in a dose-dependent manner. The IC50 of wortmannin for inhibiting migration was 10 nM, and that of LY294002 was 0.3 microM. These inhibitors also suppressed IGF-I-induced phosphorylation of protein kinase B PKB/Akt at Ser437 using concentrations that also inhibited cell motility. PD98059, an inhibitor of the MAPK pathway, was somewhat less potent than PI 3 kinase inhibitors in blocking cell migration that had been stimulated by IGF-I. When IGF-I increased migration of pSMC 2.1-fold above control, 100 nM wortmannin inhibited this response by 79%, 1 microM LY294002 inhibited it by 58%, and 50 microM PD98059 caused a 34% reduction. In comparison, 100 nM wortmannin inhibited IGF-I stimulated DNA synthesis by 57%, 1 microM LY294002 inhibited it by 59%, whereas 50 microM PD98059 suppressed it completely. Thus, activation of PI 3 kinase plays the major role in IGF-I-stimulated migration and proliferation of pSMC. While the activation of the MAPK pathway seems to be necessary for stimulation of mitogenesis by IGF-I, the contribution of this pathway in IGF-I induced cell migration is limited in pSMC. Interestingly, neither PI 3-kinase inhibitors nor PD98059 blocked the increase in alphaVbeta3 integrin affinity that followed IGF-I treatment. Therefore, although both the PI 3-kinase and MAPK pathways were used by IGF-I to increase migration of pSMC, alphaVbeta3 integrin activation did not depend on either PI 3-kinase or MAPK activation, suggesting the possible importance of some other signal transduction pathway to account for its full actions on pSMC. PMID- 10465297 TI - Maternal epidermal growth factor deficiency causes fetal hypoglycemia and intrauterine growth retardation in mice: possible involvement of placental glucose transporter GLUT3 expression. AB - We investigated the physiological role of epidermal growth factor (EGF) in fetal growth in mice in which midgestational sialoadenectomy induced maternal EGF deficiency. Sialoadenectomy decreased the fetal weight significantly, indicating that maternal EGF deficiency caused intrauterine growth retardation. The weight of the fetal liver in the sialoadenectomized mice was reduced in proportion to the decrease in body weight (82.7+/-10.2 vs. 70.9+/-10.9 mg), whereas the brain weight was not reduced. Sialoadenectomy significantly decreased the glucose concentration in fetal plasma (86.0+/-13.0 vs. 63.0+/-11.8 mg/dl) without affecting the maternal plasma level of glucose. Transplacental transfer of 3H-2 deoxyglucose was significantly decreased by sialoadenectomy (5.17+/-1.25 vs. 2.94+/-1.02%), but transfer of 14C-aminoisobutyric acid was not affected. Northern blot analysis and in situ hybridization of glucose transporter isoform GLUT1 and GLUT3 messenger RNAs (mRNAs) in placenta revealed that sialoadenectomy significantly reduced the expression of GLUT3 mRNA without affecting GLUT1 mRNA levels. Administration of anti-EGF antiserum enhanced the effects of EGF deficiency, which were almost completely corrected by EGF supplementation. These results indicate that EGF plays an important role in fetal growth by regulating the transplacental supply of glucose via GLUT3 expression in the placenta. PMID- 10465298 TI - Insulin-like growth factor I circumvents defective insulin action in human myotonic dystrophy skeletal muscle cells. AB - Primary human skeletal muscle cell cultures derived from muscles of a myotonic dystrophy (DM) fetus provided a model in which both resistance to insulin action described in DM patient muscles and the potential ability of insulin-like growth factor I (IGF-I) to circumvent this defect could be investigated. Basal glucose uptake was the same in cultured DM cells as in normal myotubes. In DM cells, a dose of 10 nM insulin produced no stimulatory effect on glucose uptake, and at higher concentrations, stimulation of glucose uptake remained significantly lower than that in normal myotubes. In addition, basal and insulin-mediated protein synthesis were both significantly reduced compared with those in normal cells. In DM myotubes, insulin receptor messenger RNA expression and insulin receptor binding were significantly diminished, whereas the expression of GLUT1 and GLUT4 glucose transporters was not affected. These results indicate that impaired insulin action is retained in DM cultured myotubes. The action of recombinant human IGF-I (rhIGF-I) was evaluated in this cellular model. We showed that rhIGF I is able to stimulate glucose uptake to a similar extent as in control cells and restore normal protein synthesis level in DM myotubes. Thus, rhIGF-I is able to bypass impaired insulin action in DM myotubes. This provides a solid foundation for the eventual use of rhIGF-I as an effective treatment of muscle weakness and wasting in DM. PMID- 10465299 TI - Direct identification of two contact sites for parathyroid hormone (PTH) in the novel PTH-2 receptor using photoaffinity cross-linking. AB - Direct examination of the interacting sites between PTH and the human PTH2 receptor (PTH2R) was conducted by photoaffinity cross-linking followed by protein digestion and mapping of the radiolabeled photoconjugated receptor. Photoreactive analogs of PTH, individually substituted with an L-p-benzoylphenylalanine (Bpa) at each of the first 6 N-terminal positions, were pharmacologically evaluated in cells stably expressing recombinant PTH2R. One highly bioactive analog, [Bpa1,Nle8,18,Arg13,26,27,L-2-Nal23,Tyr34]PTH-(1-34)NH 2 (Bpa1-PTH), was chosen for cross-linking studies. In addition, a PTH analog in which the photoreacive moiety is at the mid-region position 13 (K13) was demonstrated to be bioactive, then cross-linked to PTH2R. The minimal digestion-restricted domain containing the contact site ("contact domain") for 125I-Bpa1-PTH is in the sixth transmembrane domain and part of the third extracellular loop, spanning residues Ser364-Met395 of the receptor. This domain was further confirmed and refined by cross-linking 125I-Bpa1-PTH to two receptor mutants, PTH2R[V380M]- and PTH2R[V380M,M395L]-receptors. Treatment of the cross-linked conjugates with cyanogen bromide identified a single amino acid (position 380) as the putative contact point. The contact domain for 125I-K13 is located in the N-terminal extracellular tail of the receptor (in the C-terminal portion) and spans Gln138 Met147. Further validation of this contact domain was accomplished by photocross linking to point-mutated PTH2R[K137R] receptor. Previous studies in which PTH analogs were cross-linked to human PTH/PTHrP receptor (PTH1R) identified Met425 and Phe173-Met189 as the contact sites for Bpa1-PTH and K13, respectively. These studies demonstrate that both receptor subtypes, PTH1- and PTH2-receptors, use analogous sites for interaction with positions 1 and 13 in PTH. PMID- 10465300 TI - Kit-ligand/stem cell factor induces primordial follicle development and initiates folliculogenesis. AB - Initiation of folliculogenesis through the induction of primordial follicle development in the ovary has an important role in determining the fertility and reproductive fitness of most mammalian species. The factors that control this critical process are largely unknown. The hypothesis tested in the current study was that kit-ligand/stem cell factor (KL) promotes the initiation and progression of primordial follicle development in the ovary. Ovaries from 4-day-old rats were maintained in organ culture for 5 and 14 days and treated with no factor (control), recombinant kit-ligand (KL), or gonadotropins (FSH and hCG). Follicles in ovarian sections were counted and histologically classified as primordial (stage 0), early primary (stage 1), primary (stage 2), transitional (stage 3), or preantral (stage 4). Fresh ovaries from 4-day-old rats contained 68% primordial follicles (stage 0) and 32% developing follicles (stages 1-4) per section. After 5 and 14 days in culture, section from control ovaries contained approximately 41% and 55%, respectively, developing follicles (stage 1-4) per section due to spontaneous development of primordial follicles. Spontaneous primordial follicle development was completely blocked by ACK-2, a c-kit antibody that blocks KL actions. This observation suggests that endogenous KL is necessary for primordial follicle development in vitro. After 14 days of KL treatment, sections from ovaries contained 17% primordial follicles (stage 0) and 83% developing follicles (stage 1-4) per section demonstrating a dramatic induction of primordial follicle development by KL. Gonadotropins (FSH and hCG) did not induce primordial follicle development but did increase the percentage of preantral follicles (stage 4) per section. This small increase in preantral follicles in response to gonadotropins was blocked by ACK-2 suggesting that KL may in part mediate gonadotropin actions after the initiation of primordial follicle development. Ovaries contained an average of 309+/-10 follicles per section. The total number of follicles per section did not significantly vary between treatments suggesting that the effects of KL were not due to an alteration in follicle number (i.e. survival). KL appears to be one of the first factors identified to be involved in the promotion of primordial follicle development. Results suggest that KL is necessary and sufficient to induce primordial follicle development and initiate folliculogenesis. PMID- 10465301 TI - Simultaneous and independent visualization of the gonadotropin-releasing hormone receptor and its ligand: evidence for independent processing and recycling in living cells. AB - The first step in GnRH signaling is binding by the peptide to its plasma membrane receptor (GnRHR). The receptor is a member of the seven transmembrane G protein coupled class but lacks the characteristic C-terminal cytoplasmic tail, making it among the smallest receptors in this superfamily. It has been known since 1980 that agonist occupancy of the GnRHR results in patching, capping, and internalization, although it has not been possible to localize the unoccupied GnRHR, because elaboration of receptor antisera has not been easy to achieve. The recent production of a green fluorescent protein (GFP) conjugate of the GnRHR ("rGnRHR-C-tail-GFP") that is expressed in cells, targeted to the plasma membrane, binds GnRH analogs and couples to G proteins has made it possible to monitor movement of the unoccupied receptor by confocal microscopy. In the present study, we used this probe, along with Texas Red conjugates of a GnRH agonist, to examine simultaneous processing of the receptor and its ligands. The preparation of the GFP GnRHR chimera has been described. A Texas Red conjugate was made from the GnRH agonist D-Lys6-Pro9-des-Gly10EA-GnRH by standard procedures. Bioactivity of this conjugate was confirmed. Confocal fluorescence images of living GGH3 cells showed that the agonist binds the GFP-GnRH receptor construct on the cell membrane and causes the internalization of vesicles delimited by a membrane. Shortly after internalization, the agonist separates from receptor inside the vesicle, although it is still enclosed in membranes containing free receptor. As the vesicles approach the perinuclear space, the separation between receptor and agonist is more pronounced. Free receptor appears at the cell membrane after the internalization of agonist has been completed. The protein synthesis inhibitor, cycloheximide (1 mM) did not inhibit this process, suggesting that the free receptor results from the recycling of previously internalized vesicles rather than from newly synthesized receptor. These studies show visual evidence for recycling of the GnRH receptor in cultured cells. PMID- 10465302 TI - Calcitonin inhibits anterior pituitary cell proliferation in the adult female rats. AB - Previous studies have shown that CT-like immunoreactive peptide(s) (pit-CT) is synthesized by the anterior pituitary (AP) gland, and exogenously added salmon(s) CT inhibits PRL release and PRL gene transcription in cultured AP cells. Anti-sCT serum, which immunoreacts with pit-CT, stimulates PRL secretion, suggesting pit CT is a physiologically relevant PRL-inhibiting hormone. Using proliferating cell nuclear antigen (PCNA) staining and 5-bromo-2'deoxyuridine (BrdU) incorporation into newly replicated DNA, the effect of calcitonin (CT) on cellular proliferation in the rat anterior pituitary gland (AP) was examined. CT significantly attenuated PCNA-immunopositive as well as BrdU-positive AP cell populations in dispersed rat AP cells. A second series of experiments tested the effects of CT on AP cell proliferation in vivo. OVX + E2 rats were injected with 200 microg CT (iv), the rats killed at various time points, and the APs were processed for BrdU staining. CT inhibited BrdU incorporation at all time points up to 15 h after the injection, and this inhibitory effect was reversed at later time points. The effect of CT was concentration dependent, and a maximal inhibition was observed 10 h after the CT injection. Subsequent experiments identified CT-responsive AP cell populations using double immunofluorescence for BrdU and either PRL or FSH. The number of BrdU-labeled lactotropes in the AP gland declined by 74% in the CT-treated rats. Neutralization of endogenous pit-CT by passive immunization with anti-sCT serum caused a 2-fold increase in AP cell proliferation. These results suggest an important role for the endogenous pit-CT in regulation of lactotrope population of the AP gland. PMID- 10465303 TI - Corticotropin-releasing factor receptor expression in the pituitary of fetal sheep after lesion of the hypothalamic paraventricular nucleus. AB - Both the capacity of CRF to release ACTH and the number of binding sites for CRF in the anterior pituitary decline during the final weeks of gestation in fetal sheep. The present study examined regulation of pituitary CRF receptor expression by the hypothalamic paraventricular nucleus (PVN) during late gestation in fetal sheep. Bilateral radiofrequency lesions of the PVN (PVN-Lx; n = 4) or sham lesions (SHAM; n = 5) were performed in fetal sheep at 118-122 days of gestational age (dGA). Pituitary glands from PVN-Lx and SHAM fetuses were collected at 139-142 dGA (term, approximately 148 dGA). Dual-label in situ hybridization was performed using a digoxigenin-labeled ovine POMC complementary RNA, together with a 35S-labeled ovine CRF type I (CRF1) receptor complementary RNA, to localize and quantify CRF1 receptor mRNA in POMC-hybridizing cells. Binding of [125I]-ovine CRF was also examined in the fetal pituitary of both PVN Lx and SHAM fetuses using in situ autoradiography. The hybridization signal for the CRF1 receptor mRNA was primarily restricted to POMC-expressing cells in the anterior pituitary of both PVN-Lx and SHAM fetuses; no hybridization signal for the CRF1 receptor was observed in the neurointermediate lobe (NIL) in either group. The hybridization signal for CRF1 receptor mRNA in anterior pituitary corticotropes of PVN-Lx fetuses was significantly lower in both the inferior and superior regions of the anterior pituitary, compared with SHAM fetuses (P < 0.05). In the inferior region of the anterior pituitary, the percentage of POMC hybridizing cells containing CRF1 receptor hybridization signal was significantly greater in PVN-Lx (90+/-7%; mean +/- SEM), compared with SHAM (67+/-6%; P < 0.05) fetuses. No differences in the percentage of POMC cells containing CRF1 receptor hybridization signal were observed in the superior region of the anterior pituitary between PVN-Lx (89+/-8%) and SHAM (87+/-9%). Binding of [125I]-ovine CRF (oCRF) was significantly greater in anterior pituitaries of PVN-Lx (140+/-19 mean arbitrary densitometry U +/- SEM), compared with SHAM (73+/-23; P < 0.05) fetuses. For both PVN-Lx and SHAM fetuses, there were no differences within group in [125I]-oCRF binding between the inferior and superior regions of the anterior pituitary. A weak, but significant (P < 0.05), autoradiographic signal for [125I] oCRF binding was observed in the NIL of both SHAM and PVN-Lx fetal sheep. The level of [125I]-oCRF binding was significantly lower in the NIL, compared with anterior pituitary, for both SHAM (P < 0.01) and PVN-Lx fetuses. There were no differences in [125I]-oCRF binding in the NIL between SHAM and PVN-Lx fetal sheep. Our findings support a role for the PVN in regulating anterior pituitary CRF1 receptor expression in the late-gestation sheep fetus. PMID- 10465304 TI - Expression of transforming growth factor-beta1, activin A, and their receptors in thyroid follicle cells: negative regulation of thyrocyte growth and function. AB - Thyroid growth and function are intricately regulated by both positive and negative factors. In the present study, we have investigated the expression of transforming growth factor-beta (TGF-beta) super-family members and their receptors in normal porcine thyroid follicle cells. In tissue sections of porcine thyroids, we observed an expression of TGF-beta1, activin A, and bone morphogenetic protein (BMP)-7 proteins. The staining was localized to the follicular epithelium. In affinity cross-linking experiments, TGF-beta1 was found to bind to heteromeric complexes of TGF-beta type I and type II receptors, and activin A bound most efficiently to heteromeric complexes of activin type IB and type II receptors. We were unable to detect any BMP receptors (BMPRs) in attempts to perform affinity cross-linking with BMP-7. However, expression of BMPR-IA and BMPR-II messenger RNA (mRNA) was detected by Northern blot analysis. Both TGF beta1 and activin A, but not BMP-7, increased the phosphorylation of Smad2, induced nuclear translocation of Smad2, Smad3, and Smad4, and inhibited thyrocyte cell growth as well as TSH-stimulated cAMP response. TGF-beta1 was more potent, compared with activin A, to induce these cellular responses. Taken together, our findings indicate a role for several members of the TGF-beta family in regulation of thyroid growth and function. PMID- 10465305 TI - Lipocortin 1 (annexin 1): a candidate paracrine agent localized in pituitary folliculo-stellate cells. AB - It is now well established that lipocortin 1 (LC1) plays an important role as a mediator of early delayed glucocorticoid feedback action in the hypothalamo hypophysial system. In both the hypothalamus and anterior pituitary gland, LC1 mimics some of the actions of glucocorticoids; moreover, glucocorticoids stimulate the synthesis of LC1 and cause the translocation of intracellular LC1 to the outer cell surface. The mechanism by which LC1 acts in these tissues is only partially understood, but may involve paracrine and/or autocrine actions. To address these possibilities we have investigated the localization of LC1 in the rat pituitary gland, using double labeling immunohistochemistry to identify the pituitary cell types that express LC1. At the light microscopic level LC1 was not detected in the endocrine cells in cryosections of the pituitary, but it was found in abundance in the surrounding folliculo-stellate (FS) cells. In the anterior and interme diate pituitary lobes, there was a near total colocalization of LC1 and S100, a specific marker of FS cells. By contrast, in the posterior pituitary gland, LC1 immunoreactivity was not colocalized with S100 which labeled most pituicytes, or with OX-42 monoclonal antibody, a marker of the microglial cells. Immunogold electron microscopy confirmed that LC1 is present in the nongranulated FS cells. LC1 im munoreactivity was also present in a mouse pituitary FS-like cell line (TtT/GF), particularly in the periphery of the cytoplasm. The localization of LC1 in the FS cells of the anterior pituitary gland defines LC1 as a new marker of the FS cell population. These results support our hypothesis that LC1 acts as one of the paracrine agents liberated by FS cells that modulate the release of pituitary hormones. PMID- 10465306 TI - Expression and function of estrogen receptor subtypes in granulosa cells: regulation by estradiol and forskolin. AB - The expression and function of estrogen receptor ERalpha/beta subtypes and ERbeta variants in granulosa cells have been determined using several integrated approaches:, Western blotting, indirect immunofluorescence, RT-PCR, and transient transfection assays. Each of these approaches has provided specific details concerning the dynamics of ER expression, ER functional activity, and estradiol (E) regulation of target genes in granulosa cells. Specifically, the studies presented herein document that messenger RNAs (mRNAs) encoding ERbeta and its splice variants, as well as mRNA encoding ERalpha, are expressed in granulosa cells of immature rats before and during culture in serum-free medium. The results also provide the first documentation that functional (DNA binding and transcriptionally active) ER is present in cultured granulosa cells and that its ability to bind consensus estrogen response element (ERE) oligonucleotide and to transactivate an ERE promoter-reporter construct is associated with the level (type?) of receptor protein as well as the stage of granulosa cell differentiation. Using a labeled ERE consensus oligonucleotide and antibodies specific for ERbeta and ERalpha, we show that ERbeta but not ERalpha was detected (supershifted in electrophoretic mobility shift assays) in extracts of granulosa cells cultured overnight (0 h) in defined medium alone. When the cells were cultured with FSH and testosterone (T) to stimulate their differentiation, ERbeta binding activity, as well as immunoreactive ERbeta as determined by Western blot analyses, decreased progressively from 24 to 48 h and was undetectable by 72 h. ERbeta mRNA was low, and ERbeta binding activity was not observed in luteinized granulosa cells. ERalpha DNA binding activity was not observed in any of the granulosa cell cultures, although low levels of immunoreactive ERalpha were detected by Western blot analyses. Immunofluorescent analyses documented that ERbeta, as well as ERalpha, were localized to granulosa cell nuclei and that the intensity of nuclear staining was related to agonist stimulation and differentiation: forskolin increased, whereas E decreased immunostaining for ERbeta and ERalpha at 48 h. When an ERE-E1b-luciferase vector was transfected into granulosa cells of unprimed rats, basal luciferase activity was low but increased by forskolin (3-4x) and by E (2x), responses to both agonists being blocked by the ER antagonist, ICI. When the same vector was transfected into differentiated granulosa cells (cultured for 48 h with FSH/T), forskolin alone increased activity. Collectively, these results show that ERbeta protein is preferentially expressed in immature granulosa cells, is functionally active (binds DNA), can transactivate (either as a homodimer or heterodimer with ERalpha) ERE-containing promoter constructs, and might be associated with increased expression of the endogenous gene encoding c-Jun. PMID- 10465307 TI - Estrogen receptor immunoreactivity is present in the majority of central histaminergic neurons: evidence for a new neuroendocrine pathway associated with luteinizing hormone-releasing hormone-synthesizing neurons in rats and humans. AB - The central regulation of the preovulatory LH surge requires a complex sequence of interactions between neuronal systems that impinge on LH-releasing hormone (LHRH)-synthesizing neurons. The reported absence of estrogen receptors (ERs) in LHRH neurons indicates that estrogen-receptive neurons that are afferent to LHRH neurons are involved in mediating the effects of this steroid. We now present evidence indicating that central histaminergic neurons, exclusively located in the tuberomammillary complex of the caudal diencephalon, serve as an important relay in this system. Evaluation of this system revealed that 76% of histamine synthesising neurons display ERalpha-immunoreactivity in their nucleus; furthermore histaminergic axons exhibit axo-dendritic and axo-somatic appositions onto LHRH neurons in both the rodent and the human brain. Our in vivo studies show that the intracerebroventricular administration of the histamine-1 (H1) receptor antagonist, mepyramine, but not the H2 receptor antagonist, ranitidine, can block the LH surge in ovariectomized estrogen-treated rats. These data are consistent with the hypothesis that the positive feedback effect of estrogen in the induction of the LH surge involves estrogen-receptive histamine-containing neurons in the tuberomammillary nucleus that relay the steroid signal to LHRH neurons via H1 receptors. PMID- 10465308 TI - Parathyroid hormone-related protein in rat penis: expression, localization, and effect on cavernosal pressure. AB - Although PTH-related protein-(1-36) [PTHrP-(1-36)] is known to be expressed in smooth muscle and to exert potent myorelaxant effects, its tonic effects on cavernosal smooth muscle has not yet been explored. Using the RT-PCR technique, the present study establishes that PTHrP messenger RNA is present in microdissected corpus cavernosa in the rat. In immunohistochemical studies using affinity-purified antibodies to middle regions of PTHrP, immunostaining was localized throughout the penile structures, including vessels, cavernosal smooth muscle, and trabecular fibroblasts. Strong immunostaining for PTHrP was also detected in the dorsal nerve bundles. In anesthetized rats, intracavernosally injected boluses of increasing doses of PTHrP-(1-36) (0.3-30 pmol in 100 microl saline) had little effect on intracavernosal pressure. However, they markedly potentiated the dilatory response to papaverine (8-800 nmol), increasing the papaverine-induced intracavernous pressure by 2.5-fold, close to the mean arterial pressure. In conclusion, the cavernosal expression of PTHrP messenger RNA, the distribution of immunoreactive PTHrP throughout the structuro-functional components of the erectile apparatus and its strong potentiating action on papaverine-induced cavernosal relaxation, collectively suggest that PTHrP participates in the control of cavernosal tone. PMID- 10465309 TI - Regulation and localization of matrix metalloproteinases and tissue inhibitors of metalloproteinases in the mouse ovary during gonadotropin-induced ovulation. AB - At the time of ovulation, proteolytic degradation of the follicular wall is required to release the mature oocyte. Extracellular proteases, such as serine proteases and matrix metalloproteinases (MMPs), are thought to play important roles in this process. In this study we have examined the regulation of 11 MMPs and 3 tissue inhibitors of metalloproteinases (TIMPs) during gonadotropin-induced ovulation in the mouse. Northern blot hybridization showed that messenger RNA for several MMPs and TIMPs, including gelatinase A, MT1-MMP, stromelysin-3, MMP-19, TIMP-1, TIMP-2, and TIMP-3, were present at detectable levels in the mouse ovary. In addition, ovarian extracts contained gelatinolytic activities corresponding to the inactive proforms of gelatinase A and gelatinase B. Most of the MMPs and TIMPs were expressed at a constitutive level throughout the periovulatory period. However, MMP-19 and TIMP-1 revealed a different expression pattern; they were both induced 5-10 times by hCG and reached their maximum levels at 12 h after hCG treatment, corresponding to the time of ovulation. At this time point, MMP-19 and TIMP-1 messenger RNA were localized to the granulosa and thecal-interstitial cells of large preovulatory and ovulating follicles. This temporal and spatial regulation pattern suggests that MMP-19 might be involved in the tissue degradation that occurs during follicular rupture and that TIMP-1 could have a role in terminating MMP activity after ovulation. PMID- 10465310 TI - The proinflammatory cytokine, interleukin-1alpha, reduces glucocorticoid receptor translocation and function. AB - Proinflammatory cytokines have been shown to influence the expression and function of the glucocorticoid receptor (GR). Specifically, several studies have found that cytokines induce a decrease in GR function, as evidenced by reduced sensitivity to glucocorticoid effects on functional end points. To investigate the potential mechanism(s) involved, we examined the impact of the proinflammatory cytokine, interleukin-1alpha (IL-1alpha), on 1) GR translocation from cytoplasm to nucleus using GR immunostaining, 2) cytosolic radioligand GR binding, and 3) GR-mediated gene transcription in L929 cells stably transfected with the mouse mammary tumor virus-cholamphenicol acetyltransferase reporter gene. L929 cells were treated with IL-1alpha (100 and 1000 U/ml) for 24 h in the presence or absence of dexamethasone (Dex; 10 nM to 1 microM). IL-1alpha inhibited Dex-induced GR translocation and alone induced GR up-regulation. Pretreatment with IL-1alpha followed by Dex treatment for 1.5 h led to about 20% inhibition of Dex-induced GR-mediated gene transcription, whereas coincubation of IL-1alpha plus Dex for 24 h inhibited Dex-induced GR-mediated gene activity up to 42%. The latter effect was reversed by the IL-1 receptor antagonist. These results suggest that cytokines produced during an inflammatory response may induce GR resistance in relevant cell types by direct effects on the GR, thereby providing an additional pathway by which the immune system can influence the hypothalamic-pituitary-adrenal axis. PMID- 10465311 TI - Estrogen stimulates gene expression and protein production of osteoprotegerin in human osteoblastic cells. AB - The identity of the paracrine mediator(s) of the antiresorptive action of estrogen on bone cells is controversial. Osteoprotegerin (OPG) was recently identified as a soluble member of the tumor necrosis factor (TNF) receptor (TNF R) superfamily that is secreted by osteoblast lineage cells and acts by binding to and neutralizing its cognate ligand, OPG-L, a required factor for osteoclastogenesis. OPG prevents bone loss when administered to ovariectomized rats, induces osteoporosis when ablated in knock-out mice, and induces osteopetrosis when overexpressed in transgenic mice. In conditionally immortalized, human osteoblastic hFOB/ER-3 and hFOB/ER-9 cell lines containing physiological concentrations of approximately 800 and approximately 8,000 functional estrogen receptors (ER)/nucleus, respectively, we found that 17beta estradiol dose- and time-dependently increased OPG mRNA and protein levels to maximal levels of 370% and 320%, respectively (P < 0.001); co-treatment with the "pure" antiestrogen ICI 182,780 abrogated these effects completely. 17beta Estradiol also dose-dependently increased OPG mRNA and protein levels in normal human osteoblasts with approximately 400 ER/nucleus by 60% and 73%, respectively. Thus, estrogen enhancement of OPG secretion by osteoblastic cells may play a major role in the antiresorptive action of estrogen on bone. PMID- 10465312 TI - Functional oxytocin receptors discovered in human osteoblasts. AB - Undifferentiated or differentiated human trabecular bone cells with osteogenic capacity in primary culture express oxytocin receptors (OTRs). OTR expression then persists upon differentiation to an osteoblast phenotype. A human epithelial osteosarcoma cell line, Saos-2, also expresses OTRs. Expression was determined both at mRNA and protein levels. Functional OTRs are evidenced by an increase in intracellular calcium concentration, [Ca2+]i, in response to 10 nM oxytocin (OT). An oxytocin antagonist (OTA) blocked this effect, demonstrating specificity for OT. OT also stimulated prostaglandin E2 (PGE2) synthesis in both confluent undifferentiated and differentiated human trabecular bone cells. This is the first report of OTR mRNA and protein expression and of prescribed OT signal pathways in osteoblastic cells. Since PGE2 has been shown to increase bone turnover in favor of bone formation, OT may be a new class of a bone anabolic agent. PMID- 10465314 TI - On "Bayesian monitoring...". PMID- 10465313 TI - Bayesian monitoring of phase II trials in cancer chemoprevention. AB - Early randomized Phase II cancer chemoprevention trials which assess short-term biological activity are critical to the decision process to advance to late Phase II/Phase III trials. We have adapted published Bayesian interim analysis methods (Spiegelhalter et al., J. R. Statist. Soc A, 1994; 157: 357-416) which give greater flexibility and simplicity of inference to the monitoring of randomized controlled Phase II trials using intermediate endpoints. The Bayesian stopping rule is designed to stop the trial more quickly when the evidence suggests ineffectiveness rather than when it suggests biological activity, thus allowing resources to be concentrated on those agents that show the most promise in this early stage of testing. We investigate frequentist performance characteristics of the proposed method through simulation of randomized placebo controlled trials with a growth factor intermediate end-point using mean and variance values derived from the literature. Simulation results show expected error rates and trial size similar to other commonly used group sequential methods for this setting. These results suggest that the Bayesian approach to interim analysis is well suited for monitoring small randomized controlled Phase II chemoprevention trials for early detection of either inactive or promising agents. PMID- 10465315 TI - Surveying physicians to determine the minimal important difference: implications for sample-size calculation. AB - The minimal important difference (MID) is the smallest benefit of treatment that would result in clinicians recommending it to their patients. The MID is necessary to calculate sample size for randomized clinical trials, but its chosen value is often arbitrary. This study set out to determine the practicability of surveying physicians to elicit the MID for clinical trial sample-size calculation. Using a mail survey, we elicited the MID of different physician specialties (family medicine, internal medicine, vascular surgery) for using propranolol to slow abdominal aortic aneurysm (AAA) growth assuming that propranolol was efficacious in this condition. We used different outcome measures (growth rate or proportion of patients requiring surgery) and different methods of data presentation for the proportion of patients requiring surgery (absolute risk reduction or number needed to treat). The MID varied significantly by physician specialty, experience with AAA and propranolol, and the method used to elicit the MID. Consequently, sample-size calculations using these various MIDs varied from 116 to 3015. Future attempts to elicit the MID need to consider carefully who is surveyed, how data are presented, and how opinions are elicited. PMID- 10465316 TI - Exploring heterogeneity in meta-analysis: is the L'Abbe plot useful? AB - By using a published meta-analysis as an example, this paper discusses the use of L'Abbe plot for investigating the potential sources of heterogeneity in meta analysis. As compared with other graphic procedures, the L'Abbe plot is useful to identify not only the studies having different results from other studies, but also the study arms that are responsible for such differences. This may be important for determining the focus of heterogeneity investigations. Results of stochastic simulation indicate that, purely because of random variation, studies with event rates of around 50% are more likely to be identified as outliers in a L'Abbe plot. This paper also demonstrates that different methods may identify different trials as "outliers" in meta-analysis. PMID- 10465317 TI - Healthy volunteer effect in industrial workers. AB - Volunteers for epidemiological research, have lower mortality rates than non volunteers, thereby producing a bias referred to as the "healthy volunteer effect" (HVE). Occupationally active persons have been similarly shown to have a reduced mortality relatively to the general population (the "healthy worker effect"). To determine whether a HVE exists in occupationally active persons, we followed for 8 years a cohort of Israeli male industrial employees, of whom 71.6% agreed to participate in 1985 in screening examinations for cardiovascular disease. We calculated standardized mortality ratios (SMRs) of the entire cohort relative to the general population, and compared the mortality among participants with that of the non-participants. Over 8 years follow up, SMRs were 78% for the entire cohort, 71% for participants and 99% for non-participants. Participants were older than non-participants and worked more commonly in smaller factories. A proportional hazard model indicated that after adjusting for these variables, the all cause mortality hazard ratio among participants and non-participants was 0.69 (95% CI = 0.51-0.94). During the first and last two years of the 8-year follow-up there were 39.6 and 30.0 age-adjusted deaths per 10,000 person-years among participants, and 58.6 and 51.5 respectively among non-participants. We conclude that the HVE occurs in occupationally active persons, and that it may persist for up to 8 years follow-up. PMID- 10465318 TI - Mortality from dementia in advanced age: a 5-year follow-up study of incident dementia cases. AB - Five-year follow-up of a community-based, 77+ old cohort including incident dementia cases was used to evaluate the impact of dementia on the risk of death, taking into account other chronic conditions potentially related to death, and contrasting Alzheimer's disease (AD), and vascular dementia (VaD). In this population, 70% of the dementia cases died during the five years after diagnosis, with a mortality rate specific for dementia of 2.4 per 100 person-years. After controlling for sociodemographic variables and comorbidity, 14% of all deaths could be attributed to dementia with a risk of death among demented subjects twice as high as that for non-demented people. Mortality risk ratios were 2.0 (95% confidence interval 1.5-2.7) for AD and 3.3 (95% confidence interval 2.0 5.3) for VaD. This study confirms that dementing disorders are a major risk factor for death. Even in the oldest old (85+), dementia shortens life, especially among women. PMID- 10465319 TI - Misclassification and under-reporting of acute myocardial infarction by elderly persons: implications for community-based observational studies and clinical trials. AB - We investigated the accuracy of self-report of hospitalization for acute myocardial infarction (MI) by elderly persons in a community-based prospective study. Among 3809 persons aged 65 years or older followed up for 6 years, self reported hospitalization for MI was validated by review of primary records and Medicare diagnoses. Among 147 who self-reported MI and for whom hospital records were available, the diagnosis was confirmed in 79 (54%). Myocardial infarction was not a reason for hospitalization among the remaining 68 participants; misclassification with other cardiovascular diagnoses was common. Medicare diagnosis correlated well with primary hospital records. Using Medicare diagnoses as the standard, the diagnosis of MI was confirmed in 53% of self-reports; the sensitivity and specificity of self-report were 51% and 98%, respectively. False negative reporting was common because only half of hospitalizations for MI were reported. Self-report of hospitalization for MI by elderly persons in the community may be unreliable for ascertaining trends in cardiovascular diseases. PMID- 10465320 TI - Frailty bias in comorbidity risk adjustments of community-dwelling elderly populations. AB - Our objective was to describe distortion in outcome studies due to "frailty bias" or differential susceptibility to adverse health outcomes due to frailties but attributed to other factors. We linked an administrative database to survey data (n = 5934) containing functional, condition, and outcome measures. The disease classification scheme of an empirically derived mortality model was used to categorize 7500 ICD-9-CM codes into five risk levels. Cox and logistic regressions were used to compare outcomes. Commonly employed measures differ in their sensitivity to detect and control frailty bias across a spectrum of major chronic diseases. Survival is inversely related to increasing functional impairment after adjusting for age, race, gender, education, number of comorbid conditions, and highest disease risk occurring during follow-up. Functional status appears to be a superior and essential element for control of the frailty bias that threatens comparability of outcome measures across community-dwelling populations containing chronically-ill disabled elderly. PMID- 10465321 TI - Changing patterns of coronary heart disease in the hunter region of New South Wales, Australia. AB - A population-based observational study of men and women aged 35-69 years in the Hunter Region of New South Wales, Australia, was conducted to assess the impact of risk-factor modification and increased drug therapy on the trends in major coronary events and case fatality. From 1985 to 1993, there were 3006 coronary deaths and 6450 nonfatal major coronary events. Rates of death and nonfatal myocardial infarction declined, but there was an increase in hospital admissions for prolonged chest pain. Reductions in cigarette smoking, diastolic blood pressure, total cholesterol, and increased use of aspirin can fully explain the 3.3% (95% confidence interval [CI] 2.4, 4.2) average annual reduction in rates of major coronary events for men and the 4.1% (95% CI 2.7, 5.5) reduction for women. In contrast, increased use of aspirin, beta-blockers, fibrinolytic therapy, and angiotensin-converting enzyme inhibitors explain less than half of the 8.9% (95% CI 5.9, 11.8) and 6.9% (95% CI 2.7, 10.9) average annual reduction in case fatality in hospital for men and women, respectively. These trends suggest a decline in severity of coronary heart disease consistent with reductions in risk factor levels and improved acute medical treatment. PMID- 10465322 TI - Relationship of health-related quality of life to symptom severity in diabetes mellitus: a study in Trinidad and Tobago. AB - Our objective was to estimate the effect of greater symptom severity in diabetes mellitus on measures of health-related quality of life in a cross-sectional design in 35 government primary care health centres in Trinidad. Data were gathered on 2,117 subjects with clinical diabetes and analysed for 1,880 (89%). For each scale of the short form 36 (SF-36) questionnaire (a generic measure of health-related quality of life), scores were presented by quartile of symptom severity, measured using the Diabetes Symptom Checklist. Mean (SD) SF-36 scores were 44 (10) for the physical component score (PCS) and 45 (12) for the mental component score (MCS). Greater severity of diabetic symptoms was associated with lower scores on each of the subscales of the SF-36. Comparing lowest and highest quartiles of DSC score, the adjusted difference in PCS was -11 (95% confidence interval -12 to -9) and for MCS -16 (-18 to -14). Our results provide standardised data for health related quality of life in relation to severity of illness from diabetes, these might be used to aid the evaluation of relevant interventions. PMID- 10465323 TI - Aging, comorbidity, and reduced rates of drug treatment for diabetes mellitus. AB - Advanced age and its related comorbidity may affect both the patterns and goals of diabetes treatment. We examined the relationships of demographic variables and comorbidity with drug treatment for diabetes in the elderly. We studied both the 81,700 residents of New Jersey, aged 65-99 years, who were hospitalized between July 1, 1989 and June 30, 1991 and had prescription drug coverage either through Medicaid or the Pharmacy Assistance for the Aged and Disabled program, and a sample of 80,000 nonhospitalized elderly beneficiaries in these programs. Rates of utilization of insulin or oral hypoglycemic drugs in the 120 days before admission were substantially lower in those aged > or = 85 or in nursing homes. Among patients with previously treated and diagnosed diabetes, the likelihood of treatment after discharge declined with older age (odds ratio [OR] for treatment in those aged > or =85 relative to 65-74 years: 0.57; 95% confidence interval [CI]: 0.45-0.72), nursing home residence (OR: 0.30; CI: 0.22-0.41), and higher levels of comorbidity (OR for modified Charlson index > or = 5 relative to 0: 0.43; CI: 0.27-0.67). In patients who had a discharge diagnosis of diabetes but no prior treatment, those in nursing homes and those with greater comorbidity also had lower rates of diabetes treatment after discharge. Although the prevalence of diabetes increases with age and the risks of many consequences of diabetes remain high, the rate of drug treatment for diabetes declines with older age and greater comorbidity, perhaps because of concern about side effects or reduced treatment benefits due to competing risks of death. Absence of data from randomized clinical trials of diabetes treatment in the elderly appears to have resulted in considerable physician ambivalence on the benefits and risks of glycemic control in older diabetics. PMID- 10465324 TI - The prevention education program (PEP). A prospective study of the efficacy of family-oriented life style modification in the reduction of cardiovascular risk and disease: design and baseline data. AB - We describe design and baseline data of the Prevention Education Program (PEP), a home-based and family-oriented intervention program, aimed to assess and improve cardiovascular risk factors in school children and their families during an intervention period of 10 years. Started in 1994 in the German town of Nuremberg, currently 37 elementary schools (22 control and 15 intervention schools) are enrolled including 1740 families (1740 first graders, 3046 parents, and 1521 siblings). Major cardiovascular risk factors as well as dietary behavior are evaluated yearly using structured interview, physical examination, laboratory analysis, and seven-day-dietary protocols. The intervention package is applied to all families from intervention schools using regular home visits, health curricula and group sessions. Primary outcome is any reduction in cardiovascular risk factors by dietary intervention and health education compared to the control group getting only written information on the individual risk profile. The presented baseline data showing a high prevalence of cardiovascular risk factors in adults and in their children underline the need for such an intervention program in Germany. PMID- 10465325 TI - Influencing prescribing for urinary tract infection and asthma in primary care in Sweden: a randomized controlled trial of an interactive educational intervention. AB - Our aim was to evaluate effects on prescribing for urinary tract infection (UTI) and asthma, of an education with messages based on national guidelines, aimed at improving prescribing in primary care in Sweden. The study is part of the European Drug Education Project. A randomized controlled trial, with groups of general practitioners (GPs) allocated to education on UTI (18 groups, 104 GPs) or asthma (18 groups, 100 GPs), the two parallel intervention arms being controls for each other. Feedback was provided on the GP's judgments of simulated cases and prescribing. Prescribing indicators were developed and measured before and after the intervention. Analysis was performed by multi-level technique. Prescribing of first choice UTI drugs increased in the intervention arm from 52% to 70% and remained constant in the control arm (P < 0.001). The proportion of patients receiving an inhaled corticosteroid increased insignificantly in both study arms. The educational model can be used to improve prescribing. Further studies are needed to define when the model is effective. PMID- 10465326 TI - Verocytotoxins (Shiga-like toxins) produced by Escherichia coli: a minireview of their classification, clinical presentations and management of a heterogeneous family of cytotoxins. AB - Bacterial virulence usually requires the interaction of multiple factors in order to cause disease. The enterotoxins produced by certain strains of bacteria are proteins which vary in their mode of action, but do fall into two general groups; the cytotoxic and the cytotonic enterotoxins. While cytotoxic enterotoxins typically kill eucaryotic cells (eg. by inhibiting protein synthesis), cytotonic enterotoxins derange cell metabolism in specific ways (eg. by elevating cyclic nucleotide levels). Some strains of Escherichia coli produce protein toxins that are biologically, structurally and antigenically related to a cytotoxin (Shiga toxin) (ShT) produced by Shigella dysenteriae type 1. Although this group of related, but not necessarily identical toxins have been referred to as Vero cell toxins or Verocytotoxins (VTs), the term Shiga-like toxins (SLTs) has been widely accepted. ShT and SLTs have been implicated as a cause of diarrhoea as well as haemorrhagic colitis (HC) and haemolytic uremic syndrome (HUS) in humans, whilst SLTs have been implicated as causal agents of oedema disease and HC in weaner pigs and calves, respectively. While S. dysenteriae is an invasive organism, the SLT-producing strains of E. coli have not been reported to be invasive, but cause diarrhoea that may contain blood and mucus. Thus, SLTs can be considered an important "new" type of enterotoxins whose role in the pathogenesis of diarrhoea, HC and HUS is beginning to emerge, not only in certain geographical settings, but worldwide. This mini review focuses on this family of SLTs, because of recent advances which have been made towards their detection, nomenclature, pathogenesis and possible management of their clinical presentations. PMID- 10465327 TI - Semiliki forest virus vector carrying the bovine viral diarrhea virus NS3 (p80) cDNA induced immune responses in mice and expressed BVDV protein in mammalian cells. AB - Bovine viral diarrhea virus (BVDV) is a primary pathogen responsible for bovine enteric, respiratory and reproductive failure. A genetic region is encoding the p80 (NS3) of BVDV as the most conserved protein among Pestiviruses. BVDV infection in cattle induces NS3 specific lymphocyte proliferation and humoral responses. To generate a DNA vaccine against BVDV, the gene for BVDV-NADL NS3 was cloned into an eukaryotic expression vector of Semiliki Forest virus (pSFV-1). Quadriceps muscles of BALB/c mice were injected with recombinant DNA generated statistically significant cytotoxic T-lymphocyte activity (CTL) and cell mediated immune (CMI) responses against cytopathic and noncytopathic BVDV. Whereas, the BVDV-NS3 did not generate neutralizing antibodies against BVDVin mice. pSFV-1-NS3 DNA was subjected to in vitro transcription into mRNA. The mRNA was transfected into baby hamster kidney cells (BHK-21) and Madin-Darby bovine kidney cells (MDBK). The recombinant cells were used in the detection of DNA antigen responses by immunological assays. This report establishes the ability of BVDV-NS3 DNA inoculation to induce a strong cellular immune responses in mice. PMID- 10465328 TI - Epizootiologic background of dissimilar distribution of human cases of Lyme borreliosis and tick-borne encephalitis in a joint endemic area. AB - Lyme borreliosis (LB) and tick-borne encephalitis (TBE) are co-endemic in some parts of Europe, however, their distribution differs despite a common tick vector and comparable animal hosts. A serosurvey of game and small mammals was made in a highly endemic area and compared with historical data in human cases; the epidemiologic risk and the population density of game were modelled using a geographic information system. While LB-risk corresponded with an overall population density of game (red deer, roe deer, mouflon, wild boar) regardless of mouse abundance, TBE-risk suggested a dependence on the abundance of mice on the one hand, and game, particularly roe deer, on the other. While the prevalence of TBE-antibodies generally grew with the game's age, it was virtually constant at about 65% in LB irrespective of species. It implies a cumulation of scarce TBE infection histories during the game's lifetime, and thus a limited size of TBE foci relative to the living space of these animals, as well as omnipresent LB foci, in which the animals became continuously re-infected. More ecological prerequisites seem necessary to keep TBE circulating in nature which may be responsible for its confined distribution. PMID- 10465329 TI - Experimental model of enterotoxigenic Escherichia coli infection in pigs: potential for an early recognition of colibacillosis by monitoring of behavior. AB - The hypothesis that altered behavior is a sign for an early recognition of disease was tested. The experiment was conducted to evaluate the behavioral patterns of pigs in a model of postweaning colibacillosis. Twenty-five weaned pigs (from a herd that was previously found to be highly susceptible to F4+ Escherichia coli strains) were randomly assigned into 5 groups, kept in isolated pens under the controlled ambiental conditions. One day after weaning, the pigs from three groups were intragastrically inoculated (via orogastric tube) with either F4ac+ (1466 or 2407) or F4- (1467) nonenterotoxigenic E. coli (non-ETEC) strains, respectively. The pigs from the fourth group were inoculated with F4ac+ ETEC strain M1823 and the remaining 5 pigs that received broth containing 1.2% sodium bicarbonate were kept as noninoculated controls. The pigs were examined daily and the frequency and duration of their behavioral patterns, such as eating, drinking, lying, standing, urinating, defecating, rooting and playing were monitored for 300 h during a period of 10 days. In this model, three conditions were also observed in F4-susceptible pigs: (1) acute fatal diarrheal disease; (2) moderate diarrhea and weight loss and (3) no diarrhea and weight loss. The incidence (both frequency and duration) of defecating was significantly higher (P < 0.05) in pigs inoculated with F4ac+ ETEC strain M1823 as compared to that of noninoculated (control) pigs. Pigs inoculated with F4ac+ non-ETEC strain 1466 had a significantly lower frequency of eating (P < 0.05) and frequency/duration of drinking (P < 0.05) than did the controls. The 1466 inoculated pigs, had an increased diarrhea score, but frequency/duration of defecating was not significantly different. Pigs inoculated with F4ac+ non-ETEC strain 2407 spent more time in lying (P < 0.05) than did noninoculated pigs. Conversely, the pigs that received F4- non-ETEC strain 1467 laid shorter (P < 0.05) and ate/drank less frequently (P < 0.05) than the controls. It was concluded that the changed occurrence of defecating and eating in pigs that were inoculated with either F4ac+ ETEC (M1823) or non-ETEC (1466) strain. respectively, was consistent with the pending clinical disease, i.e. postweaning colibacillosis. PMID- 10465330 TI - Inhibitory effects of monoacylated 2-O-beta-galactosylglycerols on Epstein-Barr virus activation: the significant role of the hexanoyl chain. AB - Three series of monoacyl-2-O-beta-D-galactosylglycerols bearing an acyl chain of varying length, from C4 to C10, were studied due to their antitumor promoting effects on the activation of the Epstein-Barr virus early antigen (EBV-EA), such activation being induced by the tumor promoter 12-O-tetradecanoylphorbol-13 acetate (TPA). This study indicates that it is more the length of the acyl chain that is important for the activity, six carbon atoms resulting in maximum effect, rather than the position of the ester function and the nature of the sugar (galactose or glucose). PMID- 10465331 TI - Anti-tumor promoting activity of polyphenols from Cowania mexicana and Coleogyne ramosissima. AB - Chemical investigation on polyphenol-rich fractions of Cowania mexicana and Coleogyne ramosissima (Rosaceae) which showed significant inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O tetradecanoylphorbol-13-acetate (TPA), has led to the characterization of 10 compounds including C-glucosidic ellagitannin monomers and dimers from the former plant, and 17 polyphenols including flavonoid glycosides from the latter. The effects of individual components and their analogues with related structures on the TPA-induced EBV-EA activation were then evaluated. Among the compounds isolated from C. mexicana, two C-glucosidic ellagitannins, alienanin B and stenophyllanin A and a nitrile glucoside (lithospermoside), and among the constituents from C. ramosissima, two flavonoid glycosides, isorhamnetin 3-0-beta D-glucoside and narcissin were revealed to possess strong inhibitory effects on EVB-EA activation, the potencies of which were either comparable to or stronger than that of a green tea polyphenol, (-)-epigallocatechin gallate. These polyphenols except for nitrile glucoside, which was not tested owing to an insufficient amount, were also found to exhibit anti-tumor promoting activity in two-stage mouse skin carcinogenesis using 7,12-dimethylbenz[a]anthracene (DMBA) and TPA. PMID- 10465332 TI - Enhanced expression of ELAM-1 on endothelium of renal cell carcinoma compared to the corresponding normal renal tissue. AB - Renal cell carcinoma (RCC) has been shown to respond to an immunological therapy with tumor infiltrating lymphocytes (TIL), which accumulate in RCC at a higher density than in normal renal tissue, suggesting that there is selective tumor invasion. Since invasion of TIL into the malignant tissue is mediated by adhesion molecules, we examined the different expression of the adhesion molecule endothelial-leukocyte-adhesion-molecule-1 (ELAM-1) on endothelial cells of RCC versus normal renal tissue. For a specific quantification, the level of ELAM-1 mRNA was investigated by both semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis and referred to the content of endothelial cells in the tissue, determined by endothelium specific staining. Quantification of mRNA was evaluated by computer-aided integration. We observed a significantly lower amount of endothelial cells in RCC compared to normal renal tissue. The specific transcription rate of ELAM-1 in RCC, determined by RT-PCR was about 5.2 times that of normal tissue, while Northern blot analysis indicated an approximately 11.8 times increase. Our investigations show a significantly increased expression of ELAM-1 in tumor tissue compared to normal renal tissue, presumably caused by a higher amount of cytokines in the tumor tissue. This enhanced expression may be responsible for the high concentration of TIL in renal tumors. PMID- 10465333 TI - Immunohistochemically stained markers (p53, PCNA, bcl-2) in dysplastic lesions of the larynx. AB - The percentage of malignant transformation of laryngeal dysplastic lesions is difficult to estimate. There is a need for new histological markers which could enable more objective assessment of the premalignant stages of the larynx and help in estimation of the potential of future neoplastic progression. We performed a retrospective study to determine whether immunohistochemical staining for the proliferating cell nuclear antigen (PCNA), tumour suppressor gene protein p53 and antiapoptotic protein bcl-2 may be prognostic factors in laryngeal epithelial lesions. Staining was performed on 57 paraffin-embedded biopsies from patients with clinically detected precancerous stages of the larynx. Histopathologic examination revealed normal epithelium in six cases, mild dysplasia in 20 cases, moderate dysplasia in 18 cases, severe dysplasia in seven cases, CA in situ in four cases, papilloma in one case and CA invasivum in one case. The p53 count in mild and moderate dysplasia was 26.8 and 38.6%, respectively. This difference was statistically significant. There was significant correlation between PCNA and p53 scores. There was also a relationship between the scores of these markers and bcl-2 expression. In ten out of 45 cases of dysplastic lesions the invasive cancer developed in 4 years of follow-up. The correlation between PCNA score and malignant progression of the dysplastic lesions was on the statistical borderline. There was significant relationship between malignant transformation and age of the patients. PMID- 10465334 TI - The activity of caspase-3-like proteases is elevated during the development of colorectal carcinoma. AB - Activated caspase-3-like proteases promote apoptotic cell death by cleaving cellular substrates. Caspase-3-like activity was measured in colonic carcinomas and in matched normal colonic mucosa from 31 patients and was significantly elevated in 25/ 31 colonic carcinomas and adenomas when compared to normal mucosa (P < 0.0001). Caspase-3-like activity was much higher in normal mucosa and tumours of female subjects than of males (P < 0.0001). No correlation was obtained between caspase-3-like activity and location of the tumour, tumour grade, stage, or patient age. The marked increase in caspase-3-like activity in colorectal carcinomas may reflect an increase in the proportion of cells undergoing spontaneous apoptosis. PMID- 10465335 TI - Pattern of expression of beta-defensins in oral squamous cell carcinoma. AB - Human beta-defensin (hBD)-1 and hBD-2 are antimicrobial peptides that have been detected in certain types of epithelia, including the skin and oral epithelia. It has been suggested that bacterial infection is an important factor in the process of carcinogenesis. The expression of hBDs in oral squamous cell carcinoma (SCC) may be down-regulated. We studied the pattern of expression of hBD-1 and hBD-2 mRNA in oral (SCC) cell lines and in tumor samples obtained from four patients with oral SCC who underwent surgical resection, by reverse transcription polymerase chain reaction (RT-PCR). Human gingival epithelial (HGE) cells were used as the control. The effect of various inflammatory cytokines on hBD-1 and hBD-2 expression in the HGE cells and SCC cell lines, was also studied. hBD-1 mRNA was detected in the Ca-9, SCC-9 and HSC-4 cell lines, but not in the SAS and KB cell lines. hBD-2 mRNA was detected in all five cell lines. All four tumor samples expressed both hBD-1 and hBD-2 mRNA, although the mRNA level of each protein varied. These results indicate that SCCs in which hBD expression is downregulated, may be susceptible to bacterial infection. PMID- 10465336 TI - Differential display analysis can reveal patterns of gene expression in immortalised hepatoma cells which are similar to those observed in young adult but not old adult liver cells. AB - We used the differential display technique to examine whether there were any patterns of gene expression which were characteristic of both young adult rat liver and of immortalised rat hepatoma cell lines, but not of old adult rat liver. No genes were detected which appeared to be clearly expressed in young liver and immortalised cell lines, but not in old liver. However, 14 genes were detected in old liver which were down-regulated in young liver and the hepatoma cell lines. This observation lends support to the idea that immortalisation of malignant cells may involve, at least in some aspects, a reversal of the ageing process in these cells and that the genes involved have a recessive action. PMID- 10465337 TI - A multiplex-PCR/RFLP procedure for simultaneous CYP2E1, mEH and GSTM1 genotyping. AB - Inter-individual variation in metabolism of environmental toxicants, which is attributed to genetic polymorphism, may be a major risk factor in determining who will develop adverse health effects. This priority research area is the focus of many laboratories, and new techniques need to be developed to enhance the efficiency in generating data. We have developed and validated a new multiplex polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) procedure for simultaneous genotyping of cytochrome P450 II E1 (CYP2E1), microsomal epoxide hydrolase (mEH), and glutathione S-transferase mu (GSTM1). Enzymes from these three polymorphic genes are involved with the phase I and II metabolism of a variety of environmental toxicants. Therefore, simultaneous characterization of these genes will not only reduce costs but will increase the efficiency of data collection, thereby contributing to health risk assessment efforts. PMID- 10465338 TI - Detection of circulating testicular cancer cells in peripheral blood. AB - Patients who receive peripheral blood stem cell transplants are at risk of developing cancer recurrence due to the presence of malignant cells in the transplants. We investigated a sensitive method to detect malignant cells in the peripheral blood and peripheral blood stem cells of patients with testicular cancer using nested, reverse transcription-polymerase chain reaction (RT-PCR) to measure alpha-fetoprotein gene expression. Using this technique, a single cancer cell could be detected in 10(6) peripheral blood mononuclear cells. This is the first report of an attempt to detect circulating malignant cells in the peripheral blood of patients with testicular cancer by nested RT-PCR. PMID- 10465339 TI - Dietary zinc deficiency enhances esophageal cell proliferation and N nitrosomethylbenzylamine (NMBA)-induced esophageal tumor incidence in C57BL/6 mouse. AB - The effect of zinc deficiency on N-nitrosomethylbenzylamine (NMBA)-induced esophageal tumor formation in rats has been well documented. Our previous work showed that zinc deficiency and its associated increased esophageal cell proliferation were of paramount importance in esophageal tumor development in the NMBA-rat model. However, there has been no report concerning zinc deficiency and NMBA-induced esophageal tumor formation in mice. In this study, weanling C57BL/6 mice were fed ad libitum with either a zinc-sufficient or a zinc-deficient diet containing 3-4 ppm of zinc, and received six intragastric doses of NMBA (2 mg/kg; twice weekly for 3 weeks). The animals were sacrificed 46 weeks later after in vivo bromodeoxyuridine (BrDU) labeling followed by immunohistochemical detection of cells in S-phase. At 46 weeks, the tumor incidences in zinc-deficient mice were 57, 100, and 100% respectively, in the esophagus, forestomach and squamocolumnar junction with the glandular stomach (SCJ), as compared to 17, 39, and 67% in the corresponding tissue of zinc-sufficient mice. The difference between the two dietary groups was significant at P < 0.02 for the esophagus, and P < 0.001 for the forestomach and the SCJ. BrDU labeling revealed that the esophageal labeling index and the number of labeled cells were increased by zinc deficiency. These results support a role of increased cell proliferation in esophageal carcinogenesis in the mouse. PMID- 10465340 TI - Tumor-promoting activities of hydroquinone and 1,1-dimethylhydrazine after initiation of newborn mice with 1-methyl-1-nitrosourea. AB - To clarify the suitability of a newborn-mouse carcinogenesis assay to detect tumor-promoting activities of carcinogens, the non-genotoxic hydroquinone (HQ) and genotoxic 1,1-dimethylhydrazine (UDMH) were administered to mice during the promotion stage after treatment with 1-methyl-1-nitrosourea (MNU) (20 mg/kg body wt, single intraperitoneal injection) at day 9 after birth. Initiated males and females thus received either HQ at 0.8% in basal diet, or UDMH, at 20 mg/kg body wt once weekly by subcutaneous injection, from day 14 until the end of the experiment at 30 weeks of age. Uninitiated newborn mice, given an injection of the vehicle (0.01 M citrate buffer (pH 5.5), 20 mg/kg body wt), also received HQ or UDMH in the same way. Histopathologically, focal proliferative lesions were found in the livers of male mice and in the lungs of both male and female mice in the MNU-treated groups. HQ significantly increased the incidence and multiplicity of altered hepatocellular foci, the combined incidence of hepatocellular adenomas and carcinomas in males and the incidence and multiplicity of lung adenomas and the combined incidence of lung adenomas and carcinomas in female mice. In addition, four out of eleven MNU + HQ-treated male mice developed lung carcinomas, showing a significant elevation in multiplicity. UDMH also exhibited a tendency to increase the incidence and multiplicity of lung adenomas in female mice. Thus tumor-promoting effects of HQ or UDMH were apparently exerted in the target organs and the MNU-initiated two-stage newborn-mouse carcinogenesis assay may be useful for detection of genotoxic or non-genotoxic carcinogenicity. PMID- 10465341 TI - Evaluation of the transgenic p53+/- mouse for detecting genotoxic liver carcinogens in a short-term bioassay. AB - The transgenic p53-deficient heterozygous (p53+/-) mouse is prone to both spontaneous and induced tumors and has been proposed for use in a sensitive, short-term (6 months) assay for identifying genotoxic, multispecies carcinogens. It is not clear, however, if a short-term assay with p53+/- mice detects agents that target certain organs, in particular, the liver. In this study, we treated neonatal male p53+/- and p53+/+ mice with the genotoxic carcinogens dimethylnitrosamine (DMN), 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP), and 6-nitrochrysene (6-NC). In keeping with the methodology of the proposed short-term assay, the p53+/- mice were evaluated for tumors 7 months after treatment. Wild-type neonatal mice treated with genotoxic carcinogens are known to develop tumors within 1 year; hence, the p53+/+ animals used as controls were subjected to pathological examination at 1 year of age. Our results showed that PhIP was not tumorigenic in either group of mice. Liver tumor incidence increased significantly in the p53+/+ mice treated with DMN and 6-NC, indicating that the conditions of the bioassay were conducive to the promotion of liver tumorigenesis. On the other hand, these two chemicals failed to induce a significant increase in liver tumors in the p53+/- mice by seven months. This result suggests that a deficiency in the amount of p53 protein does not lead to accelerated liver tumorigenesis in mice, and contrasts with previous reports that show a decreased latency of tumors in non-liver targets. PMID- 10465342 TI - CD24 is a marker for human breast carcinoma. AB - CD24 is a small, mucin-type glycosylphosphatidylinositol-linked cell surface molecule expressed by neutrophils, pre B lymphocytes and certain human tumor cell lines. CD24 has been identified as a ligand for P-selectin in both mouse and human cells. We previously reported that the P-selectin-CD24 binding pathway is important for the binding of the breast carcinoma cell line KS to platelets and the rolling of these cells on endothelial P-selectin. In the present study we have analyzed the expression of CD24 on human breast carcinoma cell lines and on fresh breast carcinoma specimens using the CD24-specific antibody ML-5. Our study clearly demonstrates that CD24 is abundantly expressed on cell lines and fresh tissues of breast carcinomas. We find a differential expression of CD24 in breast carcinomas (cytoplasmic pattern) versus benign breast lesions (apical pattern). Moreover, the intensity of CD24 expression increases with the histological grade of the tumor. Thus, CD24 expression might be a useful marker for human breast carcinoma and play a role in facilitating metastasis by the interaction between tumor cells and platelets or endothelial cells. PMID- 10465343 TI - Erythrocyte superoxide dismutase activity in patients with digest cancer: adjuvant diagnosis test. AB - Previous studies showed that a common feature of tumor cells is their low of superoxide dismutase (SOD) activity. Our research proved that carcinogenic substances reduce erythrocyte SOD activity, while anticarcinogenic ones increase it and that in vitro SOD assessments revealed a direct mechanism of action. In this paper we determined erythrocyte SOD activity in patients with digestive and extra-digestive cancers. We studied eight groups of patients with five different forms of cancer (histopathologically confirmed) and three groups of volunteers. Group 1 (n = 86), patients with esophageal cancer; Group 2 (n = 94), patients with gastric cancer; Group 3 (n = 79), patients with colorectal cancer; Group 4 (n = 71), patients with hepatic cancer; Group 5 (n = 73), patients with pancreatic cancer; Group 6 (n = 85), patients with with other diseases, but of the same organs; Group 7 (n = 97), healthy volunteers; Group 8 (n = 91), hypertensive patients. We determined the erythrocyte SOD activity using the catecholamine oxidation method. Erythrocyte SOD from the first 5 groups had a mean value of 1.91 +/- 0.78 EU as compared with Group 6 which presented an activity of 6.08 +/- 1.02 (P < 0.001) and Group7 in which the activity was 6.54 +/- 1.23 (P < 0, 001). Erythrocyte SOD activity is significantly reduced (by 2-3 fold) in all groups of cancer patients studied. Being accessible, simple and rapid, erythrocyte SOD activity determination could constitute an adjuvant test in diagnosis of these types of cancer. Taking into account our previous results which prove that carcinogenic substances inhibit SOD activity by a direct mechanism, we suggest that further research is required in this exciting field. PMID- 10465344 TI - Pharmacological activities of fusaric acid (5-butylpicolinic acid). AB - This review article aims at summarizing research findings on the various pharmacological activities of fusaric acid (5-butylpicolinic acid), a mycotoxin produced by several Fusarium species which commonly infect cereal grains and other agricultural commodities. The actions of the toxin on mammals, birds, arthropods, crustaceans and plants are covered. The effects on mammals are diverse and are apparent in the nervous, cardiovascular and immune systems. Fusaric acid is toxic to some mammalian tumor cell lines. PMID- 10465345 TI - Characterization of kappa1-opioid receptor binding in human insular cortex. AB - Mesolimbic dopaminergic neurotransmission is modulated by dynorphin peptides binding to kappa-opioid receptors. The interaction between dynorphin and dopamine systems makes the kappa-opioid receptor a potential drug discovery target for the development of therapeutic agents for schizophrenia and drug abuse. This study reports the specificity and parameters of [3H]U69593 binding in the insular cortex, a representative corticolimbic area of the human brain. The results demonstrate that the radioligand [3H]U69593 labels a single population of receptors in human insular cortex with an affinity in the low nanomolar range. The pharmacological profile for inhibition of [3H]U69593 binding was determined in this brain region using drugs known to bind to mu, kappa and delta opioid receptors. The results show that kappa-opioid selective agonists and antagonists inhibit binding of this ligand in human brain with comparable affinities and rank order as previously described for rat and guinea pig brain and the cloned kappa1 opioid receptor subtype. PMID- 10465346 TI - Detection of gene deletions in Chinese patients with Duchenne/Becker muscular dystrophy using CDNA probes and the polymerase chain reaction method. AB - One hundred thirty-eight patients with Duchenne/Becker muscular dystrophy (DMD/BMD) were screened with complete cDNA probes and the multiplex polymerase chain reaction (mPCR) amplification of 18 pairs of oligonucleotide primers. Eighty-six deletions and 4 duplications were detected, the deletion frequency being 62.3%. Eighty-two deletions were detected with the two sets of primers described by Chamberlain et al. and Beggs et al, which was 95.4% of deletions detected by complete cDNA probes. Consistent with the deletion locations described previously, the deletions of dystrophin gene in Chinese individuals are clustered mainly in two high-frequency deletion regions of exons 44-52 (68.6%) of 3' side of the gene central regions and exons 1-19 (26.7%) in the 5' side. The distribution of deletions in dystrophin gene is associated with the phenotype of DMD/BMD. In the 25 cases with in-frame deletions, 15 deletions located in the region of exons 2-47 were milder BMD and intermediate patients, as the location of deletions was not the important region of the dystrophin gene. PMID- 10465347 TI - Alpha-stat calibration of indo-1 fluorescence and measurement of intracellular free calcium in rat ventricular cells at different temperatures. AB - To explore how to manage pH when calibrating Ca2+ probes at different temperatures, the dissociation constant (Kd) of indo-1 was determined both in pH stat (pH is fixed despite the temperature) and in alpha-stat (pH changes with temperature as in cells). The results showed that the Kd was much more sensitive to temperature in pH-stat than in alpha-stat, demonstrating that alpha-stat calibration should be preferred when using a Ca2+ probe to measure intracellular free calcium ([Ca2+]i) at different temperatures. Based on the calibration in situ and in alpha-stat, we showed a striking increase of [Ca2+]i from 141+/-8 nM at 30 degrees C to 218+/-22 nM at 10 degrees C in indo-1 loaded rat ventricular cells, which supports that intracellular calcium overload takes place in cardiac myocytes of non-hibernating mammals during hypothermia. PMID- 10465348 TI - Inhibition of GABA system involved in cyclosporine-induced convulsions. AB - In this study, we attempted to clarify the mechanisms mediating cyclosporine evoked convulsions. Cyclosporine (50 mg/kg, i.p.) significantly enhanced the intensity of convulsions induced by bicuculline (GABA receptor antagonist), but not those induced by strychnine (glycine receptor antagonist), N-methyl-D aspartic acid, quisqualic acid or kainic acid (glutamate receptor agonists). Bicuculline plus cyclosporine-induced convulsions were significantly suppressed by an activation of GABAergic transmission with diazepam, phenobarbital and valproate. The GABA turnover estimated by measuring aminooxyacetic acid-induced GABA accumulation in the mouse brain was significantly inhibited by cyclosporine (50 mg/kg, i.p.). When cultured rat cerebellar granule cells were exposed to 1 microM cyclosporine for 24 hr, the specific [3H]muscimol (10 nM) binding to intact granule cells decreased to 53% of vehicle controls. The present study provides the first evidence suggesting that cyclosporine inhibits GABAergic neural activity and binding properties of the GABAA receptor. These events are closely related to the occurrence of adverse central effects including tremors, convulsions, coma and encephalopathy under cyclosporine therapy. PMID- 10465349 TI - Overexpression of the G protein G11alpha prevents desensitization of Ca2+ response to thyrotropin-releasing hormone. AB - Doubly transfected human embryonal kidney cells (clone E2M11 of the HEK 293 cell line) expressing both thyrotropin-releasing hormone (TRH) receptors and G11alpha protein in high amounts were used to analyze the desensitization phenomenon of the Ca2+-mobilizing pathway. Quite unexpectedly, we did not observe any significant desensitization of the [Ca2+]i response to TRH in these cells after repeated or prolonged incubation with the hormone (up to 5 h). Under the same conditions, the TRH-induced [Ca2+]i response was completely desensitized in the parent cell line (293-E2 cels) expressing TRH receptors alone. In both cell lines, inositol phosphate response was desensitized after TRH exposure, although basal levels of inositol phospates in TRH-pretreated cells were much higher than in "naive" TRH-unexposed cells. These data suggest a significant role of the G protein G11alpha in desensitization of the Ca2+-mobilizing pathway occuring after repeated or long-term exposure of target cells to TRH-receptor agonists. PMID- 10465350 TI - Xenon induces metaphase arrest in rat astrocytes. AB - The use of xenon as an almost ideal anesthetic with very little side effects is gaining clinical acceptance, yet its effects on the cellular level are still unclear. It affects intracellular Ca2+-homeostasis but up to now no cellular event or Ca2+-signaling system has been described to be specifically sensitive to xenon. Here we report for the first time a specific effect of xenon on astroglial cells not found with another volatile anesthetic, isoflurane, nor with helium nor with N2: treatment of primary astroglial cells from embryonic rat brain with xenon induces, apart from a slight retardation of the cell cycle, a block at metaphase. Upon removal of xenon cells arrested at metaphase complete their mitosis normally. Even under continuous exposure to xenon, cells can be rescued from metaphase arrest by a small and transient increase in intracellular Ca2+; cells enter anaphase despite the presence of xenon and complete cell division, exhibiting normal rate of chromosome movement and normal chromosome separation. These results suggest that xenon interferes with some Ca2+-dependent regulatory system required for the metaphase-anaphase transition; taking into account its anesthetic effects, xenon may be also involved in the control of glia-mediated signaling transfer. PMID- 10465351 TI - Changes in plasma oxidative state with age and their influence on contractions elicited by noradrenaline in the rat tail artery. AB - The present study analyzes the changes in plasma oxidative state with age and their influence on the contractions induced by noradrenaline (NA) in endothelium denuded segments from the tail artery of 6- (young), 24- (old) and 30- (very old) month-old Sprague Dawley rats. The sensitivity (-log EC50) to NA increased with age, this increase being higher in old than in very old animals. Moreover, the maximum response (Emax) to NA did not change in old rats, whereas decreased in very old animals. We also found a progressive increase in the plasma oxidative state with age, measured as malondialdehyde (MDA) levels, that was accompanied by a decrease in the plasma antioxidative state, measured as glutathione peroxidase activity. In addition, MDA (0.5, 1 and 10 microM) potentiated the NA responses in 6-, 24- and 30-month-old rats, respectively, without affecting Emax. In young animals, catalase (1000 U/ml) or dimethylsulfoxide (7 mM), scavengers of hydrogen peroxide or hydroxyl radicals, respectively, did not modify either the contractions induced by NA in control situation or the potentiation of these responses caused by MDA. However, the superoxide anion scavenger, superoxide dismutase (SOD, 150 U/ml), completely reversed the increase in sensitivity to NA caused by MDA, without affecting NA responses in control situation. These results suggest that the increase in NA sensitivity with age could be due, at least in part, to the enhancement of plasma oxidative state during aging. In addition, in this alteration of the responses to NA caused by MDA, the generation of superoxide anions appears to be involved. This study supports the hypothesis that the enhancement of plasma oxidative state could play an important role in the increase of vascular resistance with age. PMID- 10465352 TI - Cardiomyocyte apoptosis with enhanced expression of P53 and Bax in right ventricle after pulmonary arterial banding. AB - The aim of this study is to investigate whether the cardiomyocyte apoptosis is induced after experimental right-sided pressure overload and whether the expression of proapoptotic factor is altered or not. Ten-week-old male Sprague Dawley rats were subjected to right ventricular overload by experimental coarctation of the main pulmonary artery. In pulmonary artery-banded rats, TUNEL method revealed that positive nuclei were observed in cardiomyocytes exclusively in the right ventricle, and Northern blot analysis showed that p53 mRNA level in the right ventricle was 6.2-fold higher at the day 1 than that in sham-operated rats and its level decreased gradually. Bax mRNA in the right ventricle was also increased 3.3-fold at the day 1 in pulmonary artery-banded rats and also gradually decreased. The immunohistochemical study revealed that the immunoreactivity of P53 and Bax was observed exclusively in the right ventricle of the pulmonary artery-banded group. These results demonstrated that the occurrence of TUNEL-positive cardiomyocytes in the acute pressure overload was accompanied by the enhanced expression of apoptosis inducers. It is suggested that acute pressure overload is a potent apoptotic stimulus for cardiomyocytes. PMID- 10465353 TI - Myocardial ischemia/reperfusion injury in the inducible nitric oxide synthase knockout mice. AB - Inducible nitric oxide synthase (iNOS) plays an important role in the inflammatory process of certain major cardiac disorders including myocardial infarction and allograft rejection. However, the role of iNOS in acute myocardial ischemia has not been well defined. We determined the effects of genetically disruption of the intact iNOS system on cardiac tolerance to ischemia/reperfusion injury. Adult male wild-type (WT) and iNOS knockout (KO) B6,129 mice were subjected to 20 min global ischemia and 30 min reperfusion in a Langendorff isolated perfused heart model (37 degrees C, n = 10/each group). Ventricular contractile function, heart rate, coronary flow, and leakage of intracellular enzymes (CK and LDH) were not significantly different between the groups during pre-ischemia as well as reperfusion period (P > 0.05). Myocardial infarct size was also not significantly different between WT (20.2+/-2.0% of risk area) and KO mice (23.5+/-3.8%; Mean+/-SEM, P > 0.05). However, the post-ischemic heart rate was significantly preserved in KO as compared to WT (P < 0.05). We conclude that disruption of iNOS gene does not exacerbate ischemia/ reperfusion injury in the heart. PMID- 10465354 TI - Comparative toxicity of high doses of vastatins currently used by clinicians, in CD-1 male mice fed with a hypercholesterolemic diet. AB - The CD-1 male-mouse model was employed to evaluate comparatively the toxicity of four vastatins (VTS) currently used in clinical medicine: lovastatin (LVT), simvastatin (SVT), pravastatin (PVT) and fluvastatin (FVT). Each vastatin was used orally in doses of 500 mg/Kg body weight/day, in animals with a hypercholesterolemic diet (HD) 5 days, or with a control diet (CD) 30 days. The association of high doses of VTS + HD produced a significant increase in liver weight and liver weight to body weight ratio in animals with SVT and FVT. Cholesterol (Chol) and triacylglycerols (TAG) in the liver increased significantly with FVT but not with the other VTS; Chol increased and TAG decreased in serum very significantly with FVT and SVT. The serum aminotransferases increased quite significantly with FVT but not with other VTS. In the experiment with high doses of VTS + CD, the animals receiving SVT or FVT showed a moderate loss of body weight. Liver weight and liver weight to body weight ratios were similar among all groups. Liver Chol showed a significant decrease with all VTS. Serum Chol decreased moderately with LVT and FVT. TAG in serum and liver showed a moderate decrease with all VTS. The serum aminotransferases were not modified by any vastatin. Our results indicate that high doses of VTS in male mice with a hypercholesterolemic diet result in a decreasing toxicity as follows: FVT>SVT>LVT>PVT. PMID- 10465355 TI - Protective effects of poly (ADP-ribose) synthase inhibitors in zymosan-activated plasma induced paw edema. AB - The aim of the present study was to investigate the role of poly (ADP-ribose) synthetase (PARS) in a model of acute local inflammation (zymosan-activated plasma (ZAP)-induced paw edema), in which the oxyradicals, nitric oxide and peroxynitrite, are known to play a crucial role. Injection of zymosan-activated plasma (ZAP) into the rat paw induced edema formation. The maximal increase in paw volume was observed at three hours after administration (maximal in paw volume: 1.29+/-0.09 ml). At this time point, there was a marked increase in neutrophil infiltration in the paw, as measured by an increase in myeloperoxidase (MPO) activity in the paw tissue (260+/-25 mU/100 mg wet tissue). However, ZAP induced paw edema was significantly reduced in a dose-dependent manner by treatment with 3-aminobenzamide (3-AB) or nicotinamide (NIC), two inhibitors of PARS, at 1, 2, 3, 4 hours after ZAP injection. PARS inhibition also caused a significant reduction of MPO activity. The paw tissues were also examined immunohistochemically for the presence of nitrotyrosine (a footprint for peroxynitrite formation). At 3 h following ZAP injection, staining for nitrotyrosine were also found to be localised within discrete cells in the inflamed paw tissue. Treatment with PARS inhibitor prevented the appearance of nitrotyrosine in the tissues. Our results suggest that in paw edema induced by ZAP, inhibition of PARS exert potent anti-inflammatory effects. PMID- 10465356 TI - Decreased expression of the mRNA for somatostatin in the periventricular nucleus of depression-model rats. AB - Expression of the mRNA for somatostatin (SRIF) in the periventricular nucleus (PeN), the level of SRIF in the stalk-median eminence (SME) and the concentration of growth hormone (GH) in the plasma were examined in depression-model rats in an attempt to confirm the hypothesis that SRIF neurons in the hypothalamus are hypofunctional in this model. We exposed male Wistar rats to intermittent walking stress for two weeks and then we measured their spontaneous running activity for 12 days. We divided the rats into a depression-model group and a partial-recovery group according to the spontaneous running activity of each rat after the termination of exposure to stress. Expression of SRIF mRNA in the PeN of the hypothalamus was monitored by in situ hybridization and relative levels were determined with an image analysis system. The relative level of expression of SRIF mRNA in the PeN was lower in rats in the depression-model group than in the control group and the partial-recovery group. The level of SRIF in the SME was lower and the plasma concentration of GH was higher in the depression-model group than in the other groups. Our findings suggest that reduced expression of mRNA for SRIF in the PeN might be associated with the pathophysiology of rats with this particular model of depression. PMID- 10465357 TI - Endothelin receptor antagonists reduce the pressor effects of angiotensin II into the periaqueductal gray area of rats. AB - Injection of ANGII (0.01, 0.1 and 1 nmol/rat) into the periaqueductal gray (PAG) area significantly (P<0.01) increased, in a dose-dependent manner, the mean arterial blood pressure (MAP). The increases in blood pressure induced by ANGII (1 nmol; 37 +/- 4 mmHg, n=5) were greatly reduced (>85%) by pre-administration of the ET(A) receptor antagonist FR139317 (5 nmol/rat) to the PAG area, but were unaffected by the ET(B) receptor antagonist BQ-788 (5 nmol/rat). SB209670, non selective ET(A)/ET(B) receptor antagonist, also reduced the effect induced by ANGII. These results suggest that endogenous endothelin-1, via an action on ET(A) receptors, may contribute to the pressor effects of ANGII within the PAG area of rats. PMID- 10465358 TI - Domestic radon exposure and the frequency of stable or unstable chromosomal aberrations in lymphocytes. AB - PURPOSE: To clarify the relationship between domestic radon exposure and the occurrence of chromosomal aberrations, stable translocations especially, in peripheral blood lymphocytes. MATERIALS AND METHODS: The study comprised a total of 84 nonsmoking individuals, divided into three groups according to radon concentration measurements performed in their homes: low radon concentration (<100Bq/m3, mean 67Bq/m3), medium (200-400Bq/m3, mean 293Bq/m3) or high (>800Bq/m3, mean 1737Bq/m3). Minimum residence in the present low-rise house was 10 years. The groups were matched with regard to age, gender and medical exposure. Fluorescence in-situ hybridization (FISH) was performed using chromosome paints for chromosomes 1, 2 and 4; 1500 metaphases were scored from each individual. RESULTS: Equal frequencies of translocations and also other aberrations, e.g. dicentrics and complex rearrangements, were obtained in each group. Significant correlation of translocations with age was observed, and due to the high mean age (50 years) the genome-corrected frequency of translocations was high: about one translocation in 100 metaphases. CONCLUSIONS: Chronic exposure to high concentrations of domestic radon did not increase the rate of stable or unstable chromosomal aberrations in peripheral blood lymphocytes detected by FISH chromosome painting. A strong age effect was observed. PMID- 10465359 TI - Effect of oral ZnDTPA on late effects of injected plutonium in rat. AB - PURPOSE: To reduce the long-term toxicity of 239Pu in rats by lifetime drinking of ZnDTPA solution and to investigate possible side-effects of the drug. MATERIALS AND METHODS: Male Sprague-Dawley rats received a single injection of 239Pu citrate, alone or plus oral ZnDTPA. Additional groups were administered only ZnDTPA. Late tissue changes were evaluated by post-mortem examination, X rays and histologically. RESULTS: The incidence of rat bearing osteosarcoma decreased after treatment to 35% as compared with 53% in untreated controls. The proportional incidence of osteosarcomas was reduced after ZnDTPA by more than the corresponding removal of 239Pu. Unexpectedly in the male rat, mammary tumours, mostly malignant, developed in 20% of rats that received 239Pu as compared with 0.5% in the untreated controls. After a lifetime drinking solely 3 x 10(-3) M ZnDTPA the incidence of diffuse glomerulosclerosis reached 29% as compared with 10% in controls. CONCLUSIONS: In rat, protracted oral administration of ZnDTPA reduced the incidence of osteosarcomas after injection of 239Pu, even if treatment started with a delay of 1 month. In the latter case, however, more soft tissue damage was found than after treatment beginning at 4 days post-239Pu. An increased incidence of diffuse glomerulosclerosis was observed as a side effect of oral ZnDTPA only when given continuously, alone and in high amounts. PMID- 10465360 TI - Non-proportional involvement of Chinese hamster chromosomes 3, 4, 8 and 9 in X ray-induced chromosomal aberrations. AB - PURPOSE: To study by fluorescence in situ hybridization (FISH) the involvement of Chinese hamster chromosomes 3, 4, 8, and 9, and their separate chromosome arms in X-ray-induced aberrations. MATERIALS AND METHODS: Male embryonic primary cells of Chinese hamster were used and metaphases were collected and scored at 20 h after confluent cells were exposed to 1 and 4 Gy X-rays. The frequencies and types of chromosomal aberrations involving chromosomes 3, 4, 8, and 9 were studied by FISH using arm-specific painting probes. Proportional distribution of the induced aberrations was tested on the basis of the relative lengths of chromosomes or chromosome arms studied. RESULTS: A non-proportional distribution of breaks, colour junctions as well as apparently simple dicentrics and translocations among the chromosomes studied was observed in the 4 Gy group. In general, chromosome 3 was less involved than expected and chromosome 8 was more involved than expected, and chromosomes 4 and 9 were involved as expected. A non-proportional involvement of arms in breaks, colour junctions and apparently simple dicentrics was also observed. The short arm of chromosome 3 was more involved in breaks and colour junctions than expected. The long arm of chromosome 4 was more involved in dicentrics than expected. CONCLUSIONS: Results of this study indicate a non proportional involvement of Chinese hamster chromosomes 3, 4, 8 and 9 as well as their arms in different types of aberrations following irradiation. PMID- 10465361 TI - Radioprotection of DNA by spermine: a molecular modelling approach. AB - PURPOSE: To observe and explain the sequence-dependence of DNA radioprotection by spermine. MATERIALS AND METHODS: Sequencing gel electrophoresis was used to analyse the probability of frank strand break (FSB) induction at each nucleotide site. Molecular modelling of complexes of DNA with spermine molecules and of a curved electrically null DNA has been performed. RESULTS: The effect of spermine on radiation-induced strand breakage varied significantly along the studied fragment. At low spermine concentration, some sequences were protected while others were unprotected. Molecular modelling calculations show that the most electro-negative sites are located in the minor or in the major groove of DNA. The positively charged spermine (Z=+4) should preferentially bind to such sites. When bound in the minor groove, spermine triggers a reduction of the accessibility of radiolytic attack sites to OH* radicals. This is due to induced structural modifications and to the masking of attack sites. In the case of major groove binding, no reduction of accessibility occurs. This type of binding can explain the lack of protection of sequences with electro-negative sites in the major groove. At high spermine concentration, the fragment is strongly protected. A nucleosome-like pattern of breakage with periodically distributed regions of protection was observed. Molecular modelling calculations show that the accessibility of the attack sites in a curved electrically null DNA is also periodically reduced. CONCLUSIONS: Molecular modelling of DNA-spermine complexes that takes into account the electrostatic properties of DNA, allows an explanation of the experimentally observed effects of spermine on DNA radiosensitivity. PMID- 10465362 TI - Pre-exposure of human squamous carcinoma cells to low-doses of gamma-rays leads to an increased resistance to subsequent low-dose cisplatin treatment. AB - PURPOSE: To investigate low-dose hypersensitivity to cisplatin and increased resistance at higher doses of cisplatin for the human squamous carcinoma cell line SCC-25 and its cisplatin-resistant derivative SCC-25/CP, and to examine the effects of pre- and post-treatment of SCC-25 cells with low-doses of gamma-rays on their resistance to cisplatin. MATERIALS AND METHODS: SCC-25 and SCC-25/CP cells were treated with various cisplatin concentrations (0.1 to 20 microM for 1 h) and assayed for survival using a conventional colony assay. For SCC-25, various doses of gamma-rays (5 cGy to 2.5 Gy) were given either 10 or 60 min before the cisplatin challenge dose as well as either 10 or 60 min after the cisplatin challenge dose. RESULTS: Low-dose (0.5, 0.75 and 1 microM for 1 h) hypersensitivity to cisplatin and increased resistance at higher doses was detected for the SCC-25 cell line, but not for its cisplatin-resistant derivative, SCC-25/CP. Pretreatment of SCC-25 cells with an acute low-dose of 5, 25 cGy or 1 Gy gamma-rays given 60 min before a low-dose cisplatin challenge (0.1 and 1 microM for 1 h) resulted in a significant increase in resistance (p=0.2, 0.01 and <0.001 respectively). For pretreatment of SCC-25 cells with similar low doses of gamma-rays 10 min before the challenge cisplatin dose, the increased resistance was reduced or absent and was only significantly increased for pretreatment with 25 cGy and a challenge cisplatin dose of 0.1 microM for 1 h (p = 0.02). Similar acute low-doses of y-rays given either 10 or 60 min after the challenge cisplatin dose did not increase resistance. CONCLUSIONS: The human squamous carcinoma cell line SCC-25 showed a low-dose hypersensitivity to cisplatin followed by increased resistance at higher doses. Treatment of SCC-25 cells with low-doses of gamma-rays can induce a protective effect to a subsequent low-dose cisplatin challenge. PMID- 10465363 TI - Radiation-induced apoptosis and gene expression in neonatal kidney and testis with and without protein synthesis inhibition. AB - PURPOSE: To analyse the incidence of radiation-induced apoptosis, expression of two apoptosis-related genes, Bcl-2 and p53, and post-radiation levels of cell proliferation in the neonatal rat (4-5 days old) kidney and testis. MATERIALS AND METHODS: Apoptosis was quantified in control or treated kidney or testis at 2, 4, 6, 8 and 24h after 5 Gy of whole body X-irradiation (n=4 per group). Morphology (light and electron microscopy) and DNA gel electrophoresis were used to assess apoptosis. Temporal and spatial expression of Bcl-2 or p53 were analysed using immunohistochemistry. Administration of cycloheximide (1.5mg/kg) was used to determine whether new protein synthesis had a role in induction of apoptosis. Tritiated thymidine uptake and autoradiography were used to indicate alterations in cell proliferation (radiolabel administered 1 h prior to tissue collection) or S-phase cells undergoing radiation-induced apoptosis (radiolabel administered 1 h prior to irradiation). RESULTS: Apoptosis peaked at 4 h in the testis and 6 h in the kidney and was significantly higher in the renal nephrogenic zone than in the testis (p<0.05). Mitosis was almost completely negated after irradiation in both tissues. A higher proportion (almost fivefold) of the apoptotic cells died in S phase in the kidney than in the testis. Cycloheximide negated induction of apoptosis in the kidney, and markedly decreased apoptosis in the testis. Bcl-2 expression was highest in the differentiated zone of control kidneys and increased after irradiation in the nephrogenic zone, particularly near foci of apoptosis in developing nephrons. In the control testis, Sertoli cells had moderate expression of Bcl-2. After irradiation, there was complete absence of Bcl-2 expression in apoptotic Sertoli cells, with surviving cells increasing Bcl 2 expression. Irradiated kidney had more intense nuclear p53 expression compared with controls. In the testis, p53 that was present in controls continued to be expressed in surviving cells but not apoptotic cells in radiation-treated animals. CONCLUSIONS: Unique differences can be identified between the incidence and biomolecular control of radiation-induced apoptosis in the normal neonatal kidney and testis. These results may find application for minimizing damage to these normal neonatal tissues in the development of, for example, cancer treatment regimens. PMID- 10465364 TI - Delayed lethality, apoptosis and micronucleus formation in human fibroblasts irradiated with X-rays or alpha-particles. AB - PURPOSE: To determine the yields of cell lethality and micronucleus formation measured immediately after irradiation or at delayed times in primary human fibroblasts exposed to X-rays or alpha-particles. MATERIALS AND METHODS: Primary human AG01522B fibroblasts were irradiated with X-rays or alpha-particles and subcultured for up to 30 days post-irradiation. Clonogenic survival and the yield of micronucleated cells in newly divided daughter cells were scored at various times. RESULTS: After the initial dose-dependent acute response for both endpoints there was conclusive evidence for the delayed production of de novo genomic instability in the surviving progeny. As well as the production of micronucleated cells, evidence was observed for the production of fibroblasts undergoing apoptosis as measured using conventional morphological scoring, 3'OH end-labelling of DNA fragmentation and externalization of phosphatidyl serine residues to the cell surface. For both radiations, the dose and time dependencies of the apoptotic response were similar to those for micronucleation and loss of clonogenicity. CONCLUSIONS: Genomic instability in the progeny of irradiated primary human fibroblasts was observed in the form of micronucleation, apoptosis and delayed reproductive death. An increased effectiveness of alpha-particles versus X-rays was observed for both initial and delayed responses, including apoptosis. PMID- 10465365 TI - In vitro apoptosis in peripheral blood mononuclear cells induced by low-dose radiotherapy displays a discontinuous dose-dependence. AB - PURPOSE: Cells undergoing apoptosis contribute to the regulation of activated mononuclear cells (Voll et al. 1997). Low-dose radiotherapy (LD-RT) is known to improve inflammatory symptoms, but the mechanism of action is still unclear. The aim of this study was to investigate the rate of apoptosis of peripheral blood mononuclear cells (PBMC) induced by LD-RT within the therapeutic dose range of anti-inflammatory RT. MATERIALS AND METHODS: PBMC were isolated from venous blood of ten healthy volunteers and were irradiated with single doses between 0.1 and 3.0 Gy. Apoptotic nuclei were detected by flow cytometry after propidium iodide (PI) triton staining, and apoptotic cells were detected by annexin V/PI staining and cell scatter analysis. Since apoptotic cells display increased cytoplasmatic granularity and concomitant reduced cell size, they can be distinguished from viable cells in forward/side scatter (FSC/SSC) histograms. Apoptotic PBMC were further subtyped by double staining with annexin V and directly labelled monoclonal antibodies recognizing the lineage-specific surface markers CD4, CD8, and CD19, respectively. The apoptosis rate of irradiated cells was analysed in a time and dose dependent fashion and was compared to a sham-irradiated control. RESULTS: After irradiation, a dose-dependent increase in apoptosis was observed, with a discontinuity (plateau or peak) between 0.3Gy and 0.7Gy in 9/10 donors (90%) and 59/80 samples (74%). 8/10 donors (80%) and 38/80 samples (47%) showed not only a discontinuous increase with a plateau but a relative maximum of apoptosis peaking within the dose range of 0.3 Gy and up to 0.7 Gy. CONCLUSION: LD-RT induces a relative maximum of apoptosis in PBMC in the does range between 0.3 Gy and 0.7 Gy. This may contribute to its anti-inflammatory effect observed clinically. PMID- 10465366 TI - Radiation-induced changes in nucleotide metabolism of two colon cancer cell lines with different radiosensitivities. AB - PURPOSE: To investigate changes in nucleotide metabolism after irradiation. MATERIALS AND METHODS: HT29 and SW48 human colon carcinoma cells were exposed to 60Co gamma-rays at doses ranging from 0 to 7.5 Gy. At different times after irradiation, the activities of nine enzymes involved in nucleotide metabolism were measured, the levels of thymidine kinase and deoxycytidine kinase proteins were evaluated by Western blot, and cell-cycle kinetics were analysed by flow cytometry. RESULTS: Changes in enzyme activities concerned not purine but pyrimidine metabolism and essentially the salvage pathway for deoxyribonucleotide synthesis. They were greater in the less radiosensitive HT29 cells. The levels of thymidine kinase and deoxycytidine kinase proteins changed in parallel with their activities. The metabolic changes in irradiated cells did not seem to be due to S phase transition and the pattern of enzyme activity changes was different from that observed in proliferative cells. CONCLUSIONS: Radiation-induced changes in the salvage pathway for pyrimidine deoxyribonucleotide synthesis were observed. These findings could be exploited in cancer therapy because higher enzyme activities after irradiation suggest that radiation exposure may render cells more sensitive to the drugs activated by these enzymes. PMID- 10465368 TI - Delayed cytotoxic and genotoxic effects in a human cell line following X irradiation. AB - BACKGROUND: In order to clarify the relationship between delayed reproductive death and radiation-induced genomic instability, the colony-forming efficiency of surviving, irradiated human squamous carcinoma cells and centromere positive as well as centromere negative micronuclei in surviving progeny were examined. MATERIALS AND METHODS: Colony-forming ability and micronucleus (MN) frequency in binucleated cells 24 h after the addition of cytochalasin B during 2 weeks of post-irradiation growth were determined in a squamous cell carcinoma cell line (SCL-II) of human origin. In addition, centromeres in micronuclei were detected using FISH. RESULTS: In the human epithelial cell line used for these experiments, delayed reproductive death was pronounced and persisted for at least 2 weeks after irradiation. Although there is evidence for an increased rate of centromere positive micronuclei, but not of centromere negative micronuclei, arising during the first week of post-irradiation proliferation, this decreases later while the rate of delayed reproductive death remains elevated. CONCLUSION: In the studied cell line, the observed delayed reproductive death is not closely related to the investigated criteria of radiation-induced genomic instability. This casts doubt on the common assumption that delayed reproductive death is a direct manifestation of radiation-induced genomic instability. PMID- 10465367 TI - Lethality of single-track events: comparison between calculations and experimental data. AB - Recent results on cell killing by microbeams were analysed using three parameters (k, L, L1), where k is the mean number of lethal particles per cell nucleus, L [keV/microm] is the track-average LET (linear energy transfer) in cells, and L1 [keV/microm] is a critical value for inducing lethal damage by a single track. Analysis showed that calculations are consistent with two data sets. The existence of a quadratic dependence on LET of cellular effects is confirmed in the high-LET region between 30 and 500 keV/microm. L1 approximately 150 keV/microm was found to give the best fit and the sensitive area of the cell nucleus was determined as approximately 50 microm2 for Chinese hamster V79 cells. In microbeam experiments with several MeV alpha-particles, the relationship between k and L for V79 cells can be expressed as L = approximately 150(k)-1/2. For a given survival level, the difference in the required dose (or L) between the microbeam and broad-beam experiments is also analytically described. PMID- 10465369 TI - Radiolytic and photodynamic modifications of ion transport through the plasma membrane of OK cells: a comparison. AB - PURPOSE: This study seeks to compare functional modifications of the plasma membrane induced either by water radiolysis (80 kV X-rays) or by visible light in the presence of the photosensitizer photofrin II. The two methods generate different kinds of free radicals and other reactive species such as singlet oxygen. Their effect on the electrical properties of the plasma membrane is investigated. METHOD: The patch-clamp technique (whole-cell and inside-out configuration) was applied to the plasma membrane of an epithelial cell line (opossum kidney (OK) cells). RESULTS: Generation of the reactive species gave rise to a decay of the membrane potential, to an inactivation of K+-channels, and to an increase of the leak conductance of the membrane. All three effects were strongly involved in photomodification of the membrane. Following X-ray exposure (doses up to 1000 Gy) on the other hand, the decay of the membrane conductance caused by inactivation of ion channels was predominant, while the increase of the leak conductance and the depolarization were comparatively minor responses. CONCLUSION: Photomodification of the plasma membrane by visible light in the presence of the membrane-active sensitizer photofrin II is of considerably greater consequence for the electrical properties of the membrane, compared with exposure to ionizing radiation of the membrane and its aqueous environment. The differences are thought to reflect the types of reactive species produced by the two methods and their site of generation. PMID- 10465370 TI - Topoisomerase activities and levels in irradiated Chinese hamster AA8 cells and in its radiosensitive mutant EM9. AB - PURPOSE: To investigate possible variations in topoisomerase (topo) I and II activities and levels after X-ray treatment in the radiation repair proficient AA8 Chinese hamster cell line for comparison with the radiation sensitive mutant EM9. MATERIALS AND METHODS: AA8 and EM9 cells were irradiated with 5 Gy of X-rays and the activities of topoisomerases I and II in nuclear extracts were studied. Immunological detection of both topoisomerases was carried out in order to detect any changes in the expression of these enzymes as a consequence of irradiation. RESULTS: Topoisomerase activities and levels in irradiated EM9 cells were the same as in control non-irradiated cells. In fact, both topo I and topo II activities clearly increased shortly after irradiation in the parental AA8 cells, with a more rapid increase for topo I than for topo II. In the AA8 cells, an increased level of topo I detectable immunologically was only observed at a later time (1 h) after irradiation, while no similar change was detectable for topo II. CONCLUSIONS: While this hypothesis needs further testing, an attractive idea is that DNA topoisomerases might be involved in the cellular response to radiation damage, either through a direct participation in repair mechanisms or indirectly. PMID- 10465371 TI - Radio-induced structural membrane modifications: a potential bioindicator of ionizing radiation exposure? AB - PURPOSE: This study focused on radio-induced membrane alterations in order to assess some related parameters as potential biological indicators of ionizing radiation effects in cases of accidental overexposure. MATERIALS AND METHODS: Radio-induced membrane alterations were assessed after gamma-irradiation of human blood. Biophysical techniques based on fluorescent probe incorporation into isolated living lymphocytes and erythrocytes membranes were applied. RESULTS: Using the technique of fluorescence polarization, the lipophilic phase of the membrane was shown to be more fluid whereas the lipid-protein interface of the membrane was shown to be more rigid after gamma-irradiation. Fluorescent anisotropy modifications showed dose-time effect relationships after radiation exposure. Ionizing radiation induced a decrease in steady-state anisotropy values but did not affect the probe's lifetime as assessed by fluorescence lifetime distribution technique. These data suggest that the anisotropy variations are representative of the local properties of the fluorescent probe's micro environment. However, the distribution width showed a decrease pointing towards radiation-induced changes of membrane domain organization, probably due not only to membrane water penetration related to lipoperoxidation, but also to compositional changes and redistribution of membrane components. In contrast, the lack of radiation effect observed using the lateral diffusion index technique may be related to the integrated overview of the radio-induced modifications of the membrane provided by this technique, which pointed out radio-induced damage to the membrane in micro-domains. CONCLUSION: These findings suggest the utility of structural membrane modification measurements as an early bio-indicator of ionizing radiation exposure. PMID- 10465372 TI - Fluorescent labelling of closely-spaced aldehydes induced in DNA by bleomycin Fe(III). AB - PURPOSE: To test the ability of FARPhC and FARP, two novel fluorescent reagents, to label aldehyde-containing sites (principally abasic sites) generated in DNA by the radiomimetic drug bleomycin, and to use fluorescent energy transfer from FARPhc (donor) to FARP (acceptor) to quantitate such closely-spaced sites. MATERIALS AND METHODS: FARPhc, 7-hydroxycoumarin-3-carboxylic acid (((((amino oxymethyl) carbonyl) hydrazino) carbonylethyl) amide) was synthesized with a protocol similar to the one recently reported for FARP (a fluorescein-based probe). RESULTS: Both FARPhc and FARP form stable oxime bonds with the open-chain aldehydes generated upon acidic depurination of DNA. Plasmid DNA exposed to bleomycin-Fe(III)-ascorbate undergoes extensive strand breakage, and upon subsequent reaction with FARPhc and/or FARP it becomes fluorescently labelled, indicating the generation of aldehyde-containing sites. The binding of the probes to calf thymus or plasmid DNA results in significant fluorescent energy transfer among closely-spaced fluorophores, as revealed by the fluorescence increase following digestion of fluorescently labelled samples with nuclease P1. The fluorescent quenching is most evident when both FARPhc and FARP are used simultaneously to trap aldehyde sites. When single-stranded oligonucleotides engineered to contain either one or two closely spaced bleomycin binding sites are exposed to bleomycin and then fluorescently labelled, the oligonucleotides demonstrate significantly increased fluorescent energy transfer with two binding sites indicating a dependence of aldehyde site generation and clustering on the local sequence of a single strand. CONCLUSIONS: A new detection method for DNA damage induced by bleomycin following fluorescent labelling of aldehyde group containing sites (FLAGS) and their clustering via fluorescent energy transfer is demonstrated. The method is applicable to any form of DNA. This work may lead to a general approach for the quantification of multiply damaged sites in DNA, a subset of DNA lesions that may have major biological significance. PMID- 10465373 TI - Estrogen replacement therapy in women with a history of breast cancer. AB - Estrogens are known as potent mammary mitogen substances and are the major stimulus for the growth of hormone-dependent tumors and clearly implicated in the pathogenesis of breast cancer. Therefore it is a general belief that hormone replacement therapy (HRT) after breast cancer will increase the risk of developing recurrences, though there are no clear data available to support this suggestion. No prospective study with a large number of patients and a long treatment period was performed concerning this issue. On the other hand it may not be justifiable to withhold hormone replacement therapy from low-risk patients after menopause, knowing the benefits of this therapy concerning osteoporosis and cardiovascular advantages. Nevertheless, until appropriate clinical trials help to resolve this problem, non hormonal alternatives constitute the standard of care. One possible approach is to treat menopausal women who have had breast cancer symptomatically and avoid ERT unless absolutely necessary. The risk of cardiovascular diseases can be reduced with lifestyle. Tamoxifen has a beneficial effect on serum lipids and the intake for 5 years leads to a 50% reduction in the incidence of fatal myocardial infarction and a decrease in morbidity associated with ischaemic heart disease. Low doses of progestogen is effective for menopausal hot flushes. Tibolone reduces vasomotoric symptoms such as hot flushes and offers benefit on osteoporosis and has shown a significant reduction in high density lipoprotein cholesterol. Whether replacing of estrogens is safe for patients after breast cancer remains uncertain. There is a need for a large controlled clinical trial to evaluate the safety and advantages of long time estrogen replacement in women treated for breast cancer. PMID- 10465374 TI - Hormone replacement therapy in women treated for gynaecological malignancy. AB - Can we prescribe hormone replacement therapy (HRT) safely for women, with postmenopausal complaints who were treated for a gynaecological malignancy? Only three retrospective studies have investigated this issue in endometrial cancer patients. No recurrences or deaths occurred in these treated groups. However, the physician introduced bias through the selection of favourable groups. At present, combined estrogen and progestogen therapy is probably not contra-indicated in endometrial cancer stage I and probably also not in stage II, although so far there is only circumstantial evidence. Squamous cell cancers of the cervix, vulva, and vagina are unlikely to be influenced by HRT. In the only study available of women with ovarian cancer, < or = 50 years, estrogen replacement therapy did not have a negative influence on (disease-free) survival. According to the data currently available, no evidence exists that HRT adversely influences survival and overall survival after treatment for ovarian cancer. In general, adenocarcinomas of the cervix and leiomyosarcomas of the uterus may be managed such as the adenocarcinomas of the uterus. During the last 25 years, HRT has been shown to substantially reduce the risk of cardiovascular diseases, osteoporotic fractures and colon carcinoma. On the other hand there is a significant increase of the risk in breast cancer with prolonged use of > 5 years. Re-evaluation of the current view that HRT should no be given to women treated for a gynaecological malignancy is strongly warranted after evaluating the advantages and the disadvantages of HRT use in each individual patient. Long-term HRT in women treated for a gynaecological cancer must be based on the medical history of the individual patient (and her family). PMID- 10465375 TI - Drug related negative side-effects is a common reason for poor compliance in hormone replacement therapy. AB - OBJECTIVES: The reasons for poor compliance with hormone replacement therapy (HRT) and, in particular drug-related reasons, have not yet been fully elucidated. In this study, a cohort of peri- or postmenopausal women--mainly workers from a small town and surrounding rural area--was studied. The aim of the study was to investigate why some women never start or discontinue HRT, even when great effort has been made to inform and fulfill the demands of the patient. METHODS: All women who were given a HRT prescription at a gynecological practice between September 1991 and December 1992 participated in a longitudinal study. A written questionnaire was mailed to these patients in 1996. Data from the questionnaire was supplemented with information from the medical records. Care of patients included initial information, follow-up within 4 months, yearly visits supplemented with contacts on demand. RESULTS: 356 women received the questionnaire, among which 92% replied. A total of 2% never started HRT. Seventy five percent continued the therapy for more than 3 years. Reasons for discontinuing HRT were negative side-effects (35%), desire to find out if climacteric symptoms had ended (26%), fear of cancer and thrombosis (25%), weariness of bleeding (19%) and a wish to deal with the problems 'naturally' (15%). CONCLUSIONS: Compliance with HRT can be high if adequate information is given and follow-ups are made. The main reason for poor compliance was negative side-effects, most likely progestin-related. The results of this study suggest that the future challenge will be to minimize negative side-effects of HRT. PMID- 10465376 TI - Climacteric skin ageing of the face--a prospective longitudinal comparative trial on the effect of oral hormone replacement therapy. AB - BACKGROUND: It is still a matter of debate whether HRT improves the physical quality of sun damaged skin. OBJECTIVES: To compare in a prospective longitudinal study the effects of climacteric aging controlled or not by HRT upon the tensile properties of facial skin. METHOD: A total of 140 women aged 40-52 years were enrolled in the study. The HRT group comprised 90 volunteers and a control group encompassed 50 non recipient volunteers. Yearly measurements of tensile functions of facial skin were performed for 5 years. A computerized suction device equipped with a 2-mm diameter hollow probe derived tensile variables quantifying skin distensibility, viscosity and elasticity. RESULTS: Climacteric aging was characterized by increased skin distensibility (1.1% per year) and viscosity (1.3% per year) mirrored by a decrease in elasticity (1.5% per year). HRT helped mitigate such changes. However, the HRT efficacy was not similar in all volunteers. Groups of good and poor responders were clearly identified as far as benefit on skin elasticity was concerned. CONCLUSION: The beneficial effect of HRT upon climacteric skin aging of the face is confirmed, at least in a subgroup of menopausal women. PMID- 10465377 TI - Quality of life in long-term survivors of cervical cancer. AB - OBJECTIVE: The aim of this study was to evaluate the quality of life within a sample of cervical cancer survivors. STUDY DESIGN: In this study, 46 cervical cancer survivors at clinical stage I or II and one survivor at stage 0, were evaluated. Two control groups were selected from an ongoing cohort study, in which 527 women were postmenopausal and 344 women were postmenopausal with HRT. All controls and cancer survivors completed a self-evaluating questionnaire pertaining to personal conditions, medical surveillance, individual well-being and quality of life. RESULTS: Eighteen cervical cancer survivors were ovaries preserved. A total of 28 survivors, who had an oophorectomy, were compared with the controls. Most of the cancer survivors had a good socio-economic background and an active lifestyle. Forty-two percent were currently under medical surveillance and used medication regularly. The ratio of HRT users was higher in the oophorectomy group (36% in both short- and long-term therapy). With regard to the quality of life, there were no remarkable differences between oophorectomy survivors and controls. Except that the psychological condition in the ovaries preserved group was worse, the rest of the findings concerning the quality of life derived from the ovaries preserved group were rather close to that of the oophorectomy group. CONCLUSION: The quality of life in long-term cervical cancer survivors is generally satisfactory. The positive results may be due to the higher ratio of HRT use, ovarian preservation, good education, working conditions, as well as an active lifestyle. PMID- 10465378 TI - Absorption and bioavailability of oestradiol from a gel, a patch and a tablet. AB - OBJECTIVES: To compare oestradiol and oestrone concentrations and bioavailability after a single dose and at a steady state during oral oestradiol valerate, transdermal oestradiol gel and transdermal oestradiol patch treatments. METHODS: Two open, randomised, cross-over studies were conducted. In the first study, 12 healthy postmenopausal women received 1.5 mg oestradiol as a transdermal gel or a 2 mg oestradiol valerate tablet daily for 14 days. In the second study, 15 postmenopausal women were treated for 18 days with 1.5 mg oestradiol gel or a transdermal patch releasing oestradiol 50 microg/24 h (replaced every 72 h). Venous blood samples for serum oestradiol and oestrone measurements with RIA were taken until 24 or 72 h after the first and last doses. RESULTS: The tablet and the transdermal gel yielded similar serum oestradiol profiles with a peak concentration 4-5 h after administration. The patch resulted in relatively stable oestradiol levels during the mid third of the wearing time whereas much lower levels were observed in the beginning and towards the end. There was no difference in the fluctuation between the peak and trough oestradiol levels between the gel (56 or 67%) and the tablet (54%) while the fluctuation was greater with the patch (89%). The bioavailability of oestradiol from the gel was 61% as compared with the tablet and 109% as compared with the patch. The gel was not bioequivalent with the tablet or the patch. CONCLUSIONS: The doses used of the transdermal gel and the patch roughly corresponded to each other with regard to the amount of oestradiol absorbed whereas the bioavailability from the tablet was significantly higher than from the gel. The lack of bioequivalence, the different serum oestradiol profiles and the large intersubject variability suggest that individual dose adjustments may be needed when changing administration form. PMID- 10465379 TI - Spatial distribution of NA+ and K+ channels in spinal dorsal horn neurones: role of the soma, axon and dendrites in spike generation. AB - Spinal dorsal horn neurones play an important role in processing sensory information received from primary afferent fibers. The application of the patch clamp technique to thin slices of rat spinal cord has enabled the study of ionic channels in visually identified dorsal horn neurones. The small soma of these neurones isolated from the slice by means of a novel method of 'entire soma isolation' has become a convenient model for investigating the properties and distributions of ionic channels. The present review summarizes results of recent experiments studying different types of voltage-gated Na+ and K+ channels expressed in dorsal horn neurones. Uneven distribution of the channels between the soma. axon and dendrites appears to play a major role in determining the neuronal excitability. The contribution of the soma, axon and dendrites to generation and propagation of the action potentials in central neurones is discussed. PMID- 10465380 TI - The concept of a calcium sensor in transmitter release. AB - The discovery was made in the 1940s that calcium is required for transmitter release at synapses, raising the question of the identity of the sensor molecule upon which this calcium acts. Subsequently it was shown in the 1960s that this calcium acts on the inside of the nerve terminal. The channels which mediate the influx of calcium ions into the nerve terminal were identified in the 1970s. This essay is concerned with tracing the development of the concept of a calcium sensor in nerve terminals and of recent work that identifies the sensor molecule as synaptotagmin. PMID- 10465381 TI - Subunit- and site-specific pharmacology of the NMDA receptor channel. AB - N-Methyl-D-aspartate (NMDA) receptor channels play important roles in various physiological functions such as synaptic plasticity and synapse formation underlying memory, learning and formation of neural networks during development. They are also important for a variety of pathological states including acute and chronic neurological disorders, psychiatric disorders, and neuropathic pain syndromes. cDNA cloning has revealed the molecular diversity of NMDA receptor channels. The identification of multiple subunits with distinct distributions, properties and regulation, implies that NMDA receptor channels are heterogeneous in their pharmacological properties, depending on the brain region and the developmental stage. Furthermore, mutation studies have revealed a critical role for specific amino acid residues in certain subunits in determining the pharmacological properties of NMDA receptor channels. The molecular heterogeneity of NMDA receptor channels as well as their dual role in physiological and pathological functions makes it necessary to develop subunit- and site-specific drugs for precise and selective therapeutic intervention. This review summarizes from a molecular perspective the recent advances in our understanding of the pharmacological properties of NMDA receptor channels with specific references to agonists binding sites, channel pore regions, allosteric modulation sites for protons, polyamines, redox agents, Zn2+ and protein kinases, phosphatases. PMID- 10465382 TI - Laying hens in large flocks in a perchery system: influence of stocking density on location, use of resources and behaviour. AB - 1. ISA Brown laying hens (3000) were housed in a perchery in 10 pens, each with 300 birds. The pens varied in size to produce 4 different stocking densities: 9.9 birds/m2 (n = 3), 13.5/m2 (n = 2), 16.0/m2 (n = 2) and 19.0/m2 (n = 3). Observations began at 20 weeks of age and continued until 69 weeks to establish the spatial distribution of the birds, usage of the different resources and the expression of behaviour. 2. Overall, birds spent most time on the perch frame (47%), litter area (23%), slatted floor (17%) and nestbox area (9%). 3. There was no effect of density on the proportion of birds observed on the slatted floor or on the elevated perches but as density increased the proportion on the littered area decreased. 4. Space usage was determined vertically, horizontally and longitudinally. Individual birds were seen to use about 80% of the pen volume available to them. This value was similar for all densities and showed that individuals did not have separate home ranges. 5. Fewer vertical movements were made within the main perch frame at the upper than at the lower levels but movements between the perches of the main frame and the nestbox rails were relatively frequent. This may help birds move up and down through the main frame. 6. Behaviours which decreased in incidence with crowding included moving, foraging and dust-bathing. Behaviours which increased with crowding included standing. Behaviours which were unaffected included resting, preening, prelaying behaviour, comfort behaviour and the minor behaviours. 7. The proportion of birds engaged in feeding and drinking was unaffected by density, except each time the chain feeders (which operated intermittently) ran more hens were seen feeding at the lower densities. This suggests that food delivery stimulated feeding behaviour; there may have been some restriction at the higher densities on birds feeding when and where they wanted. 8. Stocking density had no effect on the frequency of agonistic interactions: threats, lunges, comb/head pecks, chases and fights. 9. The incidence of damaging pecking was low and not density dependent. 10. Increasing density within the range investigated inhibited the expression of a number of behaviours and limited the use of specific resources: bird welfare at 19 birds/m2 may have been very slightly impaired. PMID- 10465383 TI - Ability of laying hens to jump between perches: individual variation and the effects of perch separation and motivation on behaviour. AB - 1. This study investigated the ability and the behaviour of laying hens jumping between perches 2 different distances apart using hungry and satiated birds to determine whether behaviour and success were related to motivation. 2. Four groups of 10 birds (Lohmann brown) trained to jump from one perch to another were subjected 4 times to each of the experimental treatments (2 perches with a distance of either 50 or 150 cm between them) whilst either hungry or satiated, controlling for time of day and order of presentation. 3. The behaviour of each bird on the 1st perch was observed for a maximum of 2 min or until the bird jumped from the perch. The outcome of each jump, number of head movements and steps, incidence and duration of calling, and activity score were recorded. 4. The probability of birds jumping was less at the greater distance and was not affected significantly by other experimental factors. 5. Head movements were more frequent in birds which jumped, at the shorter distance, in motivated (hungry) birds and in the morning rather than the afternoon. 6. More stepping was performed by birds which jumped and at the shorter distance; there was no significant effect of motivation or time of day on stepping rate. 7. Birds called more at the greater distance and this was interpreted as indicative of frustration. 8. Subjective scores for activity were lower when perches were separated by the greater distance and when birds were satiated (less motivated) rather than hungry. 9. Agitated head movements and stepping activity thus occurred mainly when birds were motivated and on the point of jumping whereas calling was associated with an apparent inability or unwillingness to jump. PMID- 10465384 TI - Reflective properties of domestic fowl (Gallus g. domesticus), the fabric of their housing and the characteristics of the light environment in environmentally controlled poultry houses. AB - 1. The light intensity and spectral power distribution in 3 broiler and 6 layer houses were sampled and converted to a measure which takes account of the domestic fowl's spectral sensitivity (termed lux corrected). Light intensity was highly variable around the layer houses (mean 33.2, s.d. 37.7 lux (corrected)) and to a lesser extent in the broiler houses (mean 5.2, s.d. 2.2 lux (corrected)). The mean intensities were also very much lower than on a sunny, June day at midday at Silsoe (42,937 lux (corrected)) and of different power distributions. 2. The reflectivity of the plumage of 15 traditional breeds of domestic fowl was surveyed. Most feathers showed a characteristic pattern of reflectivity, increasing either side of a wavelength of lambda = 400 nm. In 13 breeds the tail and wing feathers were darker than the breast and back feathers. Reflectivity at lambda = 700 nm was a good predictor of reflectivity at lambda = 320 nm. Food, bedding, wood and skin all had similar reflectivities to feathers. Metal, plastic and rubber all had more constant reflectivities across the spectrum. 3. The reflectivities of the fresh, feathered carcases of 3 male broilers (Cobb) and 3 layer hens (ISA Brown) were measured. There were small changes in hue and saturation within and between individual carcases, even for the apparently white broiler chickens. 4. Photographs were taken, with and without a UV(A)-only pass filter, of a broiler cock (Cobb), layer hen (ISA Brown) and cock jungle fowl. No additional patternation was evident in the UV(A)-only photographs and the jungle fowl lost most of its ornamentation. The texture of bare skin was enhanced in the UV(A)-only photographs. 5. The implications of these results for poultry behaviour are discussed. PMID- 10465385 TI - Growth of the digestive organs in ducks with considerations on their growth in birds in general. AB - 1. Growth of the oesophagus, proventriculus, gizzard, intestine, liver and pancreas weight was investigated in Mallards, White Pekins, Muscovies and a Muscovy x White Pekin cross. The birds varied in age between hatching and 154 d. The data were analysed by fitting both the Janoschek growth curve and the allometric formula. 2. The growth rate of all organs, except the oesophagus, peaked earlier (30 d for Muscovies and 14 d for the other breeds) than body weight and they grew faster to any given percentage of their final weight. In contrast, oesophagus weight showed growth curve characteristics similar to body weight. 3. Oesophagus weight showed simple, slightly negative allometry. The remaining organs followed complex allometry that can approximately be described by 2 allometric stages. The 1st phase was isometric to positive allometric. The 2nd showed marked negative allometry. 4. These growth patterns are assumed to be generally present in birds. PMID- 10465386 TI - Inbreeding and inbreeding depression on reproduction and production traits of White Leghorn lines selected for egg production traits. AB - 1. The rate of inbreeding and its effect on reproduction and egg production traits were studied in White Leghorn lines selected for egg production traits. The experiment was carried out for 10 generations in a control line (C) and in lines selected for increased egg number (EN), egg weight (EW) and egg mass (EM). 2. Data were available on reproduction traits, such as percent fertile eggs (PF), percent hatched of fertile eggs (PHF) and percent hatched of total eggs set (PHT), and on egg production traits such as age at 1st egg (AFE), egg number and egg weight. 3. The rate of increase in average inbreeding per generation was 1.50, 1.24, 1.14 and 0.18% for the line EN, EW, EM and C, respectively. The effect of inbreeding on reproduction and production traits was estimated by including the inbreeding coefficient of the hen (Fh), embryo (Fe) and mate (Fs) as a partial linear regression in the model. 4. There was a significant effect of inbreeding on reproduction traits in line EW attributable to the inbreeding of the hen, embryo and mate. No such effect was observed in the other lines. 5. In all lines inbreeding tended to reduce egg number and delay sexual maturity. In general, all lines reacted differently to inbreeding. PMID- 10465387 TI - Single-step divergent selection for male comb shape in two White Leghorn lines. AB - 1. Divergent selection for comb shape (SH, the way that the cockerel bears the comb) was performed in 2 White Leghorn lines in a study aimed at assessing possibilities for improving SH. 2. Line A, selected for large comb size (CS) at 29 weeks of age, had great SH problems whereas Line H, selected for high hyaluronic acid concentration in the comb (HA), had minor SH problems. Multivariate analyses were used to estimate genetic parameters for SH, CS and HA in lines A and H and in a control line (C). 3. Significant direct selection responses for SH were achieved in both lines. There were significant unfavourable correlated responses for CS in both lines. 4. The correlated responses for HA were not significant and were unfavourable in Line A and favourable in Line H. 5. Heritabilities for SH and CS were high in both lines and relatively low for HA. Most of the genetic correlations were in agreement with the correlated responses obtained. PMID- 10465388 TI - Inheritance of the barred plumage pattern of the Silver Campine Fowl, together with its relationship to other patterned fowl. AB - 1. An investigation was conducted among the progeny from a cross between a Silver Campine male and a Silver Spangled Hamburgh bantam female into the inheritance of the autosomal transverse barring of the Campine. 2. Both parental breeds possess the marble chickdown phenotype which has been shown to depend upon homozygosity of 3 genes; the birchen allele E*R at the E-locus, the eumelanin restrictor DB*B and the feather pattern arranging gene PG*P whilst the spangled feather pattern of the Silver Spangled Hamburgh has been attributed to homozygosity of E*R, DB*B, ML*M and PG*P, where ML*M is the eumelanin extension melanotic. 3. Examination in F1 and F2 generations of both chickdowns and adult plumage demonstrate that of the 4 loci, segregation occurs only for ML*M and hence the genotype of the Silver Campine to be homozygous E*R, DB*B, ML*N and PG*P. 4. The relationships between the genotypes of the Campine and those of other breeds with autosomal transversely barred plumage and between the Campine and those of other patterned fowl based on the E*R allele are discussed and presented in tabular form. In addition patterned fowl based on alleles other than E*R at the E-locus are included in order to demonstrate the effect of substituting E*R. PMID- 10465389 TI - Genetic-economic evaluation of traits in a goose meat enterprise. AB - 1. Goose can be considered as an additional and inexpensive meat source, provided that the marketing age does not exceed 8 weeks. Using the ability of geese to eat grass may reduce the intake of concentrated food up to 30%. 2. According to an equation developed, growth rate accounts for about 58% of the annual breeding gains, egg number 28%, feather yield 10%, fertility and mortality about 2%. These values are about the same for a wide range of food prices. 3. Employing realistic values for expected annual genetic gains reveals that the customary practice of keeping breeders for 5 to 6 years should be replaced by a much shorter cycle of 3 years because the economic gain from the shorter generation interval of selection exceeds the replacement costs. PMID- 10465390 TI - Measuring the mechanical stiffness of an eggshell using resonant frequency analysis. AB - 1. A new method was introduced for the measurement of mechanical eggshell characteristics based on the analysis of the vibration response of a chicken egg excited with a non-destructive impact. Four major topics were examined. 2. The 3 dimensional vibration pattern of an intact chicken egg was described for the spherical vibration with the lowest resonant frequency. 3. The resonant frequency of this basic vibration mode and the total egg mass were used to calculate a dynamic stiffness value of the eggshell based on a mathematical mass-spring model. 4. A correlation analysis between this dynamic stiffness value and several common egg and eggshell variable was calculated. Pearson correlation coefficients between the dynamic stiffness and static stiffness, eggshell thickness at the equator, eggshell width and shape index were all significantly different from zero and were 0.71, 0.60, 0.43 and 0.51, respectively. 5. Graphical analysis of the static vs. the dynamic eggshell stiffness indicated that eggs with thick shells (>360 microm) had a higher static stiffness value than the dynamic stiffness measurement, whereas the static stiffness of eggs with thin shells (>300 microm) was lower than the dynamic stiffness. 6. The new vibration test method is a promising alternative for evaluating mechanical eggshell properties because of its non-destructive nature and brief measuring time. PMID- 10465391 TI - Growth pattern and carcase development in male ducks selected for growth rate. AB - 1. Growth patterns of the whole body, eviscerated carcases, breast muscle, leg and thigh muscles and abdominal fat pads were compared in 4 lines (Lines A, B, C, and D) of male ducks selected for market weight (n = 1305) using growth curve analysis, allometric growth analysis and repeated measure analysis. At 49 d of age, Line A was heaviest, followed by Line B, Line C and Line D. 2. Ducks were fed ad libitum under 24-h lighting and 12 or 24 ducks were killed to determine body, carcase, breast-muscle, leg and thigh-muscle, and abdominal fat weights at time points from hatching until 53 d of age. 3. The Weibull function was chosen for growth curve analysis. The asymptote and inflection point from the Weibull growth curves identified 3 lines (Lines B, C, and D) with discrete body and carcase growth patterns but did not distinguish Line A from Line B. In all 4 lines the asymptote ranged from 4437 g to 3008 g for body weight and from 3334 g to 2098 g for carcase weight; the inflection point ranged from 22.5 d to 25.3 d for body weight and from 25.4 d to 29.6 d for carcase weight. 4. The allometric growth coefficient, relative to whole-body growth, was higher than 1.00 for breast muscle and lower than 1.00 for leg and thigh muscles during from 4 d to 53 d of age. 5. Body fat accumulation was estimated by abdominal fat. Line D accumulated more abdominal fat than other lines. The pattern of fat accumulation in Line D was different from Lines A, B and C and there were no differences between Lines A, B and C. PMID- 10465392 TI - Organic matrix composition and ultrastructure of eggshell: a comparative study. AB - 1. The avian eggshell is a biomineralised composite ceramic consisting of calcium carbonate embedded in an organic matrix. Matrix components are supposed to be involved in the control of mineralisation, crystallographic texture and biomechanical properties of eggshell. 2. The structure and eggshell matrix composition of various domesticated bird species were compared to gain insight into the universality of the eggshell mineralisation process. 3. The SDS-PAGE profiles of soluble eggshell matrix were specific within groups of birds (a: laying hen, breeder hen, quail, pheasant and possibly turkey; b: guinea fowl; c: duck and goose) but some of the protein bands were common to all groups. 4. Analogies between species were confirmed by Western blotting using hen protein antibodies. Ovocleidin-17 (OC-17) and ovalbumin were revealed in all species (except quail for OC-17). Lysozyme was present only in hen eggshell. Another egg white protein: ovotransferrin showed a positive signal in hens, turkey and quail. Osteopontin was observed in laying and breeder hens and quail. 5. Different proteoglycans were localised to discrete regions within the eggshell. Dermatan sulphate was observed within the matrix of the calcified shell of all species except quail which contained chondroitin-6-sulfate. Keratan sulphate was observed in mammillary bodies of breeder and laying hen, quail, pheasant and turkey while chondroitin sulphate was also present in guinea fowl and duck. 6. The general structural organisation of the different avian eggshells was similar but specific differences were observed in the ultrastructure of the mammillary layer. Species of the same taxonomic family could be grouped according to their structural analogies: breeder hen, turkey and pheasant resembled that of the domestic fowl. Guinea fowl was unique. Goose and duck were quite similar with large and confluent mammillary bodies. 7. Some matrix components are therefore common to eggshells of various species but more information is needed to relate differences in matrix composition between taxonomic groups with differences in ultrastructure. PMID- 10465393 TI - Effects of muscle pH and chilling on development of PSE-like turkey breast meat. AB - 1. Early post-mortem pH was used to identify PSE-like turkey meat, characterised by a low water-holding capacity and pale colour. In a commercial turkey processing facility, the pH value of turkey breast muscle was measured at 0, 9, 10, 14, 154, 231, and 246 min post mortem. 2. At 14 min post mortem, the carcases were separated into 3 pH classes: low (<5.70), medium (5.70 to 6.18), and high (>6.18). Low pH carcasses (<5.70) had higher drip loss(1.75%) than medium (0.81%) and high pH carcases (0.75%). Furthermore, high pH carcases (>6.18) had a lower L* value (darker) than low and medium pH carcases. 3. The effects of using CO2 'snow' on turkey breast muscle quality were also investigated. Light CO2 'snow' treatment (57 g snow/kg breast) and heavy CO2 treatment (113 g snow/kg breast) resulted in higher drip losses than the control no CO2 snow) group. PMID- 10465394 TI - Ideal amino acid profile and dietary lysine specification for broiler chickens of 20 to 40 days of age. AB - 1. The aim of the study was to determine the ideal ratio of the essential amino acids lysine (Lys), methionine (Met), threonine (Thr), tryptophan (Trp), arginine (Arg), valine (Val) and isoleucine (Ile) and to assess the required dietary lysine concentration for optimum performance in broiler chickens of 2 commercial strains from 20 to 40 d of age. 2. An identical basal diet was used throughout all experiments. It consisted mainly of maize and soyabean meal and contained 172 g crude protein and 13.2 MJ AME(N) per kg. For each experiment, the basal diet was adequately supplemented with all essential amino acids except the one to be tested, which was supplemented in 6 graded concentrations in exchange for maize starch. One (Met, Trp, Arg, Val, Ile) or 2 (Lys, Thr) growth trials were conducted per amino acid tested and the response in weight gain, food: gain ratio, breast meat yield and abdominal fat were examined. 3. The ideal amino acid ratio relative to Lys (expressed as a percentage) was calculated to be 75% Met+Cys, 63% Thr, 19% Trp, 112% Arg, 71% Ile and 81% Val on a true faecal digestible basis when the data were subjected to broken-line regression analysis. From both lysine studies the concentration for optimum food: gain ratio was calculated, by exponential regression analysis, to be 11.5 g true faecal digestible lys per kg diet. PMID- 10465395 TI - A comparison of ileal digesta and excreta analysis for the determination of amino acid digestibility in food ingredients for poultry. AB - 1. The apparent ileal and excreta digestibilities of amino acids in 15 samples representing 12 food ingredients were determined using 5-week-old male broiler chickens. The ingredients included 3 samples of cereals (wheat, maize and sorghum), 6 samples of plant protein meals (soyabean meal, cottonseed meal, canola meal and sunflower meal) and 6 samples of animal protein meals (meat meal, meat-and-bone meal, feather meal and fish meal). 2. The test ingredients were incorporated as the sole source of dietary protein in assay diets. Each diet was offered ad libitum to 3 pens (4 birds/pen) from d 35 to d 42 post-hatching. Total collection of excreta was carried out during the last 4 d. All birds were killed on d 42 and the contents of the lower half of the ileum were collected. Apparent ileal and excreta amino acid digestibilities were calculated using acid-insoluble ash as the indigestible marker. 3. The influence of site of measurement was found to vary among food ingredients, among samples within an ingredient and among different amino acids within an ingredient. Ileal amino acid digestibility values were similar in some ingredients, but significantly lower or higher in others than the corresponding excreta values. 4. Average ileal and excreta amino acid digestibilities in sorghum and maize were similar, but significant differences were observed for individual amino acids. In contrast, ileal amino acid digestibility values were higher than the corresponding excreta digestibility values in wheat. 5. The average ileal and excreta digestibilites of amino acids in the 3 soyabean meal samples were similar although small, but significant differences were noted for individual amino acids. Site of measurement had no effect on the digestibility of amino acids in canola meal. Digestibilities of valine, isoleucine, phenylalanine, histidine, glutamic acid, alanine and tyrosine in sunflower meal and those of valine, methionine, isoleucine, leucine, lysine, glutamic acid and alanine in cottonseed meal were lower by excreta analysis. 6. Digestibilities in animal protein meals, with the exception of blood meal and fish meal, were consistently higher by excreta analysis. Ileal-excreta differences in individual amino acid digestibilities were more evident in feather meal, meat meal and meat-and-bone meal. 7. Threonine and valine were the indispensable amino acids that were more frequently influenced by the site of measurement. Of the dispensable amino acids, aspartic acid, serine, glutamic acid and alanine were the most affected. 8. Differences determined between ileal and excreta digestibilities in the present study clearly demonstrate that amino acid metabolism by hindgut microflora in chickens may be substantial and that digestibilities measured in the terminal ileum are more accurate measures of amino acid availability than those measured in the excreta. PMID- 10465396 TI - Efficiency of utilisation of energy from maize- and broken rice-based diets in old White Leghorn and Rhode Island Red laying hens. AB - 1. The efficiency of utilisation of metabolisable energy (ME) for maintenance (k(m)) from diets containing maize and broken rice (BR) at 500 g/kg was studied in old White Leghorn (WL) and Rhode Island Red (RIR) laying hens using the respiration calorimetry technique. The maize-based diet contained 180.8 g crude protein (CP)/kg and 16.4 MJ gross energy (GE)/kg while the BR-based diet contained 173.2 g CP/kg and 16.3 MJ GE/kg. Diets were fed for 10 d, while an energy and nitrogen metabolism study was conducted during 3 d on an ad libitum fed diet followed by another 3 d on two-thirds of the ad libitum-fed quantity. 2. ME values for the maize- and BR-based diets for WL hens were 73.3% and 77.6% of the GE, whereas for the RIR hens these were 77.7% and 80.0%, respectively. 3. Fasting heat productions, determined at the end of 24 h fast for WL and RIR hens were 473.2 and 366.1 kJ/kg W0.75/d, respectively. During fasting WL and RIR hens utilised body energy reserves with efficiencies of 84.9% and 73.7%, respectively. 4. The k(m) of maize- and BR-based diets for the WL hens were 81.6% and 79.6%, whereas for the RIR hens these were 74.2% and 76.0%, respectively. 5. ME for maintenance of WL and RIR hens were 589 and 499.6 kJ/kg W0.75/d, respectively. 6. It is concluded that although WL and RIR hens differ significantly in energy metabolism, their efficiency of utilisation of energy from maize- and BR-based diets are similar. PMID- 10465397 TI - Evaluation of jojoba meal as a potential supplement in the diet of broiler breeder females during laying. AB - 1. This study was undertaken to investigate whether jojoba meal can be used as a food supplement during the laying period of chickens. 2. The size of eggs laid were smaller and the overall production rate was lower compared to control birds on food without jojoba meal supplementation. Furthermore, both ovary and oviduct weights were lower in jojoba fed birds. 3. This lowering of egg size and production rate was caused by factors present in jojoba which interfere with follicle growth, yolk deposition, progesterone production and the follicular maturation processes, resulting in the ovulation of smaller follicles and a lower ovulation rate. PMID- 10465398 TI - Hepatic concentration of heat shock protein 70 kD (Hsp70) in broilers subjected to different thermal treatments. AB - 1. The relationship between repeated thermal treatments and hepatic synthesis of Hsp 70 was studied in broiler chickens. 2. Sixty broilers were submitted to 5 different treatments (12 birds each) from day 1 to day 42. Four groups were kept in a thermoneutral environment and subjected to 0, 1, 2 and 3 heat stress episodes at 35 degrees C for 4 h per week (TN-0, TN-1, TN-2 and TN-3, respectively). The last group (HT-35) was reared at a room temperature of 35 degrees C. 3. From 39 to 42 old, the birds experienced acute heat stress at 41 degrees C. Resistance to heat stress was evaluated by the time taken for rectal temperature to increase by 3 degrees C above the pre-treatment value. Livers were collected (before and after heat stress) and Hsp70 was determined using Western Blot analysis with monoclonal anti-Hsp70 antibody. 4. Resistance to heat stress and concentration of Hsp70 were higher in those birds subjected to more heat stress episodes during the experimental period (TN-3) and HT-35. A positive correlation was observed between Hsp70 concentration and the time taken for a 3 degrees C increase in rectal temperature (r = 0.42; P<0.01). 5. Exposing birds to episodes of heat stress (35 degrees C) during rearing may improve their resistance to acute heat stress, but the previous thermal history did not seem to influence the hepatocyte Hsp70 content after exposure to more severe heat stress (41 degrees C). PMID- 10465399 TI - Length of the plateau and pipping stages of incubation affects the physiology and survival of turkeys. AB - 1. The hypothesis tested was that accelerating the rate at which a turkey embryo passes through the Plateau and Pipping stages (incubation days 25 to 28) affects growth and embryonic mortality. 2. Eggs from 4 genetic lines were incubated similarly until 24 d of incubation. The eggs were divided at random, half incubated at 36.8 degrees C (CON) and the remaining half incubated at 37.3 degrees C (FAST). 3. The passage time for each developmental stage was recorded at 4 h intervals. Tissues were collected and measured for growth, and glycogen concentration at each developmental stage in response to the accelerated development. 4. The FAST treatment accelerated passage through the Plateau in some lines but not others, but time to pip was shortened in all genetic lines. Embryonic survival rate was affected differently by FAST. Slower growth in response to FAST enhanced survival. 5. In conclusion the rate at which genetically different embryos grow, mature and survive may involve choices among the 3 processes. PMID- 10465400 TI - Corticosterone plasma concentration in male mule ducks: effects of sampling sites, repeated samplings and ACTH injections. AB - 1. Changes in plasma corticosterone concentrations according to puncture sites and various challenges including injections of an ACTH agonist (Immediate Synacthen) were investigated in male mule ducks. 2. Lower concentrations were measured in samples drawn by puncture from the occipital sinus than at a wing vein site. 3. Immobilisation and a single intramuscular injection of saline solution (1 ml, 0.9%) had no effect on plasma corticosterone after 15 min. 4. A single intramuscular injection of ACTH (5 microg/kg body weight) produced a rise (P<0.05) in corticosterone. Maximum concentrations were measured after 10 min and, in the absence of further sampling, a return to initial levels was observed by 1 h. 5. On the other hand, repeated bleedings following ACTH challenge maintained higher corticosterone concentrations. 6. A single intramuscular injection of ACTH at doses ranging between 0.625 to 20 microg/kg body weight increased corticosterone concentrations (P<0.05) in a dose-dependent manner, with the responses plateauing at doses of 1.25 microg/kg and higher. PMID- 10465401 TI - Lysine requirements of growing emus. AB - 1. The lysine requirement of growing emus between 23 and 65 d of age was determined according to growth response variables. 2. The optimal lysine requirement of emus was found to be 0.83 and 0.90 g/MJ ME for growth rate and gain:food ratio respectively. These findings are in accordance with the recommended value of 0.80 g/MJ ME, but is lower than the recommended value for ostriches (1.02 g/MJ ME) and higher than determined values for broilers (0.75 g/MJ ME) of the same age range. PMID- 10465402 TI - Synthesis and Pharmacology of the enantiomers of the potential atypical antipsychotic agents 5-OMe-BPAT and 5-OMe-(2,6-di-OMe)-BPAT. AB - The optically pure enantiomers of the potential atypical antipsychotic agents 5 methoxy-2-[N-(2-benzamidoethyl)-N-n-propylamino]tetralin (5-OMe-BPAT, 5) and 5 methoxy-2-{N-[2-(2,6-dimethoxy)benzamidoethyl]-N-n-propylamino}t etralin [5-OMe (2,6-di-OMe)-BPAT, 6] were synthesized and evaluated for their in vitro binding affinities at alpha1-, alpha2-, and beta-adrenergic, muscarinic, dopamine D1, D2A, and D3, and serotonin 5-HT1A and 5-HT2 receptors. In addition, their intrinsic efficacies at serotonin 5-HT1A receptors were established in vitro. (S) and (R)-5 had high affinities for dopamine D2A, D3, and serotonin 5-HT1A receptors, moderate affinities for alpha1-adrenergic and serotonin 5-HT2 receptors, and no affinity (Ki > 1000 nM) for the other receptor subtypes. (S)- and (R)-6 had lower affinities for the dopamine D2A and the serotonin 5-HT1A receptor, compared to (S)- and (R)-5, and hence showed some selectivity for the dopamine D3 receptor. The interactions with the receptors were stereospecific, since the serotonin 5-HT1A receptor preferred the (S)-enantiomers, while the dopamine D2A and D3 receptors preferred the (R)-enantiomers of 5 and 6. The intrinsic efficacies at the serotonin 5-HT1A receptor were established by measuring their ability to inhibit VIP-induced cAMP production in GH4ZD10 cells expressing serotonin 5-HT1A receptors. Both enantiomers of 5 behaved as full serotonin 5-HT1A receptor agonists in this assay, while both enantiomers of 6 behaved as weak partial agonists. The potential antipsychotic properties of (S)- and (R)-5 were evaluated by establishing their ability to inhibit d-amphetamine induced locomotor activity in rats, while their propensity to induce extrapyramidal side-effects (EPS) in man was evaluated by determining their ability to induce catalepsy in rats. Whereas (R)-5 was capable of blocking d amphetamine-induced locomotor activity, indicative of dopamine D2 receptor antagonism, (S)-5 even enhanced the effect of d-amphetamine, suggesting that this compound has dopamine D2 receptor-stimulating properties. Since both enantiomers also were devoid of cataleptogenic activity, they are interesting candidates for further exploring the dopamine D2/serotonin 5-HT1A hypothesis of atypical antipsychotic drug action. PMID- 10465403 TI - Thapsigargin analogues for targeting programmed death of androgen-independent prostate cancer cells. AB - A number of analogues of thapsigargin, a selective inhibitor of the sarco endoplasmic reticulum Ca2+-ATPases have been synthesized. In all of the prepared analogues the butanoyl residue at O-8 has been replaced with a residue containing an aromatic amine. The amine can be used as an anchoring point for attaching a peptide group sensitive to the proteolytic enzyme, prostate specific antigen, secreted by prostate cancer cells. Like thapsigargin, the analogues are capable of elevating the cytoplasmic Ca2+ concentration approximately sevenfold when tested at effective cytotoxic doses. The analogues in which the 8-O-butanoyl group has been replaced with 3-(4-aminophenyl)propanoyl or 4-aminocinnamoyl were found potently to induce programmed cell death of the prostate cancer cells. PMID- 10465404 TI - Synthesis and in vitro evaluation of novel 2,6,9-trisubstituted purines acting as cyclin-dependent kinase inhibitors. AB - Novel C-2, C-6, N-9 trisubstituted purines derived from the olomoucine/roscovitine lead structure were synthesized and evaluated for their ability to inhibit starfish oocyte CDK1/cyclin B, neuronal CDK5/p35 and erk1 kinases in purified extracts. Structure activity relationship studies showed that increased steric bulk at N-9 reduces the inhibitory potential whereas substitution of the aminoethanol C-2 side chain by various groups of different size (methyl, propyl, butyl, phenyl, benzyl) only slightly decreases the activity when compared to (R)-roscovitine. Optimal inhibitory activity against CDK5, CDK1 and CDK2, with IC50 values of 0.16, 0.45 and 0.65 microM, respectively, was obtained with compound 21 containing a (2R)-pyrrolidin-2-yl-methanol substituent at the C-2 and a 3-iodobenzylamino group at the C-6 of the purine. Compound 21 proved cytotoxic against human tumor HeLa cells (LD50-6.7 microM versus 42.7 microM for olomoucine, 24-h contact). Furthermore, unlike olomoucine, compound 21 was effective upon short exposure (LD50= 25.3 microM, 2-h contact). The available data suggest that the affinity for CDKs and the cytotoxic potential of the drugs are inter-related. However, no straightforward cell cycle phase specificity of the cytotoxic response to 21 was observed in synchronized HeLa cells. With the noticeable exception of pronounced lengthening of the S-phase transit by 21 applied during early-S in synchronized HeLa cells, and in striking contrast with earlier reports on studies using plant or echinoderm cells. olomoucilnc and compound 21 were unable to reversibly arrest cell cycle progression in asynchronous growing HeLa cells. Some irreversible hlock in GI and G2 phase occurred at high olomoucine concentration, correlated with induced cell death. Moreover, chmronic exposure to lethal doses of compound 21 resulted in massive nuclear fragmentation, evocative of mitotic catastrophe with minour amounts of apoptosis only. It was also found that olomoucine and compound 21 reversibly block the intracellular uptake of nuicleosides with high efficiency. PMID- 10465405 TI - Design, synthesis and biological evaluation of selective boron-containing thrombin inhibitors. AB - Based on the structural comparison of the S-1 pocket in different trypsin-like serine proteases, a series of Boc-D-trimethylsilylalanine-proline-boro-X pinanediol derivatives, with boro-X being different amino boronic acids, have been synthesised as inhibitors of thrombin. The influence of hydrogen donor/acceptor properties of different residues in the P-1 side chain of these inhibitors on the selectivity profile has been investigated. This study confirmed the structure-based working hypothesis: The hydrophobic/hydrophilic character of amino acid residues 190 and 213 in the neighbourhood of Asp 189 in the S-1 pocket of thrombin (Ala/Val), trypsin (Ser/Val) and plasmin (Ser/Thr) define the specificity for the interaction with different P-1 residues of the inhibitors. Many of the synthesised compounds demonstrate potent antithrombin activity with Boc-D-trimethylsilylalanine-proline-boro-methoxypropylglycine++ + pinanediol (9) being the most selective thrombin inhibitor of this series. PMID- 10465406 TI - Exploring the conformation of bilirubins with natural and unnatural analogues: use of positional and bridged isomers of bilirubin IXalpha. AB - Unlike bilirubin IXalpha (1), the isomers bilirubin IXdelta (2) and neobilirubin IXbeta (3) do not require conjugation with glucuronic acid in order to be excreted. A conformational analysis employing an optimized Monte Carlo method and a mixed Monte Carlo stochastic dynamics reveals that isomer 2 exhibits a structure more closed than the well known 'ridge-tile' conformation of 1. The change in the position of both propionic acid chains causes the loss of at least four hydrogen bonds. On the other hand, the change in the configuration of the distal dipyrrinone and the blockage of the lactamic nitrogen by the presence of a bridge in isomer 3 results in an open and more elongated structure, where the chance of hydrogen bond formation in this region is obliterated. The resulting molecular models for these compounds are consistent with 1H NM R, UV-vis, and TLC data. PMID- 10465407 TI - Synthesis and biological activity of high-affinity retinoic acid receptor antagonists. AB - This article reports the synthesis and biological activity of new high affinity retinioic acid receptor (RAR) antagonists. The effect of introducing heteroatoms in the bicyclic ring system of the potent dihydronaphthalene RAR antagonist 8, and the variation of the pendant aromatic group on the ability of these compounds to function as RAR antagonists is discussed. The use of binding, transcriptional, and in vivo assays revealed that the 2,2-dimethylthiochromene analogue 59, and the 2,2-dimethylchromene derivative 85, were the most effective in blocking retinoid agonist induced activity. PMID- 10465408 TI - Synthesis and evaluation of inhibitors of transthyretin amyloid formation based on the non-steroidal anti-inflammatory drug, flufenamic acid. AB - A light scattering-based amyloid fibril formation assay was employed to evaluate potential inhibitors of transthyretin (TTR) amyloid fibril formation in vitro. Twenty nine aromatic small molecules, some with homology to flufenamic acid (a known non-steroidal anti-inflammatory drug) were tested to identify important structural features for inhibitor efficacy. The results of these experiments and earlier data suggest that likely inhibitors will have aromatic-based structures with at least two aromatic rings. The ring or fused ring system occupying the outermost TTR binding pocket needs to be substituted with an acidic functional group (e.g. a carboxylic acid) to interact with complimentary charges in the TTR binding site. The promising TTR amyloid fibril inhibitors ranked in order of efficacy are: 2 > 4 approximately 7 > 3 > 9 > 6 > 21. PMID- 10465409 TI - Synthesis and preliminary pharmacological evaluation of thiophene analogues of viloxazine as potential antidepressant drugs. AB - A series of eight thienyloxymethylmorpholines, thiophene analogues of viloxazine, have been synthesized by three different routes. The preliminary pharmacological evaluation of this series shows antidepressant properties on the mice models used with a light sedative action. The structure-activity relationship is established in a first approximation. PMID- 10465410 TI - Enhanced RNA binding of dimerized aminoglycosides. AB - Aminoglycoside antibiotics have recently emerged as an intriguing family of RNA binding molecules and they became leading structures for the design of novel RNA ligands. The demystification of the aminoglycoside-RNA recognition phenomenon is required for the development of superior binders. To explore the existence of multiple binding sites in a large RNA molecule, we have synthesized covalently linked symmetrical and nonsymmetrical dimeric aminoglycosides. These unnatural derivatives were compared to their natural "monomeric" counterparts in their ability to inhibit the Tetrahymena ribozyme. The dimeric aminoglycosides inhibit ribozyme function 20 to 1.2 x 10(3) fold more effectively than their natural parent compounds. The inhibition curves of dimeric aminoglycosides have characteristic shapes suggesting the presence of at least two high affinity binding sites within the ribozyme's three-dimensional fold. The interaction of a dimeric aminoglycoside with two complementary sites of the RNA molecule is proposed. This binding motif may have implications on the development of new drugs targeting pivotal RNA molecules of bacterial and viral pathogens. PMID- 10465411 TI - Spectroscopic detection of endovesiculation by large unilamellar phosphatidylcholine vesicles: effects of chlorpromazine, dibucaine, and safingol. AB - Endovesiculation by large unilamellar vesicles (LUVs) induced by cationic amphiphiles is described in this work. A recent procedure to monitor phagocytosis of vesicles by macrophages by determining the amount of the simultaneously internalized water_soluble fluorescent dye HPTS with external quencher was adapted to LUVs (Daleke, D. L.; Hong, K.; Papahadjopoulos, D. Biochim. Biophys. Acta 1990, 1024, 352). Compared to dibucaine and safingol, the local anesthetic chlorpromazine (CPZ) was found to be the most efficient inducer of HPTS internalization by LUVs. Control experiments using LUVs with entrapped HPTS indicated that the observed dye-internalization does not originate from transient lysis. A strong increase in activity above the critical micelle concentration of CPZ implies the importance of CPZ-micelles for endovesiculation. The significantly less efficient CPZ-induced HPTS-internalization by LUVs with 68 nm compared to 176 nm diameter further diminishes the likelihood of a micelle/bilayer fusion mechanism and supports the presence of 'zipper-type' endovesiculation by LUVs with diameters as small as 68 nm. PMID- 10465412 TI - Probing the altered specificity and catalytic properties of mutant subtilisin chemically modified at position S156C and S166C in the S1 pocket. AB - A series of chemically modified mutants (CMMs) of subtilisin B. lentus (SBL) were generated employing the combination of site-directed mutagenesis and chemical modification. This strategy entails the mutation of a selected active site residue to cysteine and its subsequent modification with a methanethiosulfonate reagent CH3SO2S-R, where R may be infinitely variable. The present study was undertaken to evaluate the changes in specificity and pH-activity profiles that could be induced by modification of S156C and S166C in the S1 pocket of SBL with a representative range of side chain modifications, namely R=-CH3, -CH2C6H5, CH2CH2NH3+ and CH2CH2SO3 . The side chain of S156C is surface exposed and well solvated while that of S166C points into the pocket. Kinetic evaluation of the CMMs with suc-AAPF-pNA as substrate showed that the kcat/K(M)s changed very little for the S156C CMMs, but varied by up to 11-fold for the S166C CMMs. pH Activity profiles were also determined, and showed that a negatively or positively charged side chain modification increased or decreased respectively, the pKa of the catalytic triad histidine for both modification sites but with more dramatic changes for the interior pointing S166C than for the solvent exposed S156C site. As an additional probe of altered specificity, inhibition of the CMMs by a representative series of 5 boronic acid transition state analogue inhibitors was determined. The K(I)s observed ranged from a 3.5-fold improvement over the WT value, to a 12-fold decrease in binding. Overall, greater variability in all the parameters measured, activity, pKa, and boronic acid binding resulted from modification at the inward pointing 166 site than at the solvent-exposed 156 site. PMID- 10465413 TI - A proposed molecular model for the interaction of calcineurin with the cyclosporin A-cyclophilin A complex. AB - Cyclosporin A (CsA) and FK506 are potent natural product immunosuppressants that induce their biological effects by forming an initial complex with cytosolic proteins termed immunophilins. These drug immunophilin complexes then bind to and inhibit the serine/threonine protein phosphatase calcineurin (CN). Two classes of immunophilin have been identified with cyclophilins (CyP's) being proteins specifically binding CsA and FKBPs specifically binding FK506. Solution and crystal structures of various CsA-CyP and FK506-FKBP complexes have been determined and show no apparent structural similarity between the two classes of drug protein complexes. These findings raise the question as to how, given their structural differences, these two complexes can both inhibit CN. While the crystal structure of the FK506-FKBP12-CN complex has been reported, no structure for a CsA-CyP CN complex has been determined. Here are reported studies that use various modelling strategies to construct a model for the interaction of the cyclosporin A- cyclophilin A complex with calcineurin. The first stage of constructing this model consisted of using conformational comparison of CsA and FK506, GRID and GROUP analysis and restrained molecular dynamics to dock CsA into the FK506 binding site of the FK506-FKBP12-CN structure. An initial model for the CsA-CyPA-CN complex was then constructed by superimposing the structure of the CsA-CyPA complex onto the docked CsA molecule. This model was then optimised with molecular dynamics simulations run on sterically clashing regions. The validity of the model for the CsA-CyPA-CN complex was then examined with respect to the effect of chemical modifications to CsA and amino acid substitutions within CyPA on the ability of the drug-immunophilin complex to inhibit calcineurin. PMID- 10465414 TI - Synthesis of phosphono analogues of dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. AB - The present paper describes the synthetic routes of six phosphono analogues of dihydroxyacetone phosphate and five phosphono analogues of glyceraldehyde 3 phosphate through alpha-, beta- and gamma-hydroxyphosphonate esters precursors containing a protected carbonyl group. In some situations, depending on the sequence used for the deprotection of the phosphonate and carbonyl groups, the aldol/ketol rearrangement allowed the synthesis of either dihydroxyacetone phosphate or glyceraldehyde 3-phosphate analogues from the same precursors. All these analogues are of interest both as active-site probes and as potential substrates for glycolytic enzymes such as fructose 1,6-diphosphate aldolases (EC 4.1.2.13). PMID- 10465415 TI - Conformational considerations in the design of dual antagonists of platelet activating factor (PAF) and histamine. AB - Following the discovery of the first dual antagonist of platelet-activating factor (PAF) and histamine, 1-acetyl-4-(8-chloro-5,6-dihydro-11H benzo[5,6]cyclohepta[1,2-b]pyridin- 11-ylidene)piperidine, Sch 37370, 1, a related series of structures, exemplified by (+/-)-1-acetyl-4-(8-chloro-5,6 dihydro-11H-benzo[5,6]-cyclohepta[1,2-b] pyridin-11-yl)piperazine, Sch 40338, 2, were prepared. Interestingly, the compounds exhibited a parallel structure antiallergy activity relationship, suggesting that the two series may adopt a common conformation at the PAF receptor. Conformational analysis led to a proposal for this bioactive conformation accessible to both series. The synthesis of novel conformationally constrained analogues that might mimic the proposed bioactive conformation of these compounds, and the evaluation of their in vitro antiallergy activity form the subject matter of this report. PMID- 10465416 TI - Synthesis of E- and Z-pyrazolylacrylonitriles and their evaluation as novel antioxidants. AB - A facile synthesis of (Z)- and (E)-2-(5-arylpyrazol-3-yl)-3-(pyrrol-2 yl)acrylonitriles and (Z)-2-(1,3-diarylpyrazol-5-yl)-3-pyrrol-2 yl)acrylonitriles, and isomerisation of (Z)-2-(5-arylpyrazolyl)acrylonitriles to (E)-2-(5-arylpyrazolyl)acrylonitriles under basic conditions have been reported. (Z)-2-(1,3-Diarylpyrazolyl)acrylonitriles did not undergo isomerisation under the similar conditions. New compounds were identified on the basis of their spectral data (1H-, 13C-, 1H-1H COSY, NOESY, NOE, HMQC NMR, IR, UV and EI mass). The structures of one acrylonitrile and five of their precursor 6-arylpyran-2-ones and cyanomnethylpyrazoles were confirmed by X-ray crystallographic studies. Effects of pyrazolylacrylonitriles and their precursors on rat liver-microsomal lipid peroxidation were evaluated in vitro with a view to establish structure activity relationship and to identify a lead compound. PMID- 10465417 TI - Three-dimensional pharmacophore hypotheses for the locust neuronal octopamine receptor (OAR3). Part 2: agonists. AB - Three-dimensional pharmacophore hypotheses were built from a set of 43 agonists against octopamine receptor class 3 (OAR3) in locust nervous tissue. Among the 10 chemical-featured models generated by program Catalyst/Hypo, a hypothesis including hydrogen-bond acceptor (HBA), hydrophobic (Hp), and hydrophobic aliphatic (HpA1) features was considered to be important and predictive in evaluating OAR3 agonists. While the ideal and null hypotheses had a cost of 156.40 and 239.20, respectively, the 10 resulting hypotheses possessed costs from 169.89 to 175.81. The best hypothesis that was confirmed to have a 95% chance of true correlation yielded a low RMS of 0.757 and high regression r of 0.933. Active agonists mapped well onto all the features of the hypothesis such as HBA, Hp, and HpA1. On the other hand, inactive compounds were shown to be difficult to achieve the energetically favorable conformation which is found in the active molecules in order to fit the 3-D chemical feature pharmacophore models. PMID- 10465418 TI - Structure-requirements of isocoumarins, phthalides, and stilbenes from Hydrangeae Dulcis Folium for inhibitory activity on histamine release from rat peritoneal mast cells. AB - We examined the structure-activity relationships of isocoumarins, phthalides and stilbenes isolated from Hydrangeae Dulcis Folium and related compounds for the inhibition of histamine release in rat peritoneal mast cells. The activities of isocoumarins such as thunberginols A and B were more potent than those of dihydroisocoumarins such as hydrangenol and thunberginol G. The double bond at the 3-position seemed to be essential to potentiate the activity. The hydroxyl groups at the 8-, 3'- and 4'-positions of isocoumarin were essential for the activity, while the hydroxyl group at the 6-position was scarcely needed. Since the activities of benzylidenephthalides such as thunberginol F were more potent than those of hydramacrophyllols A and B, the presence of a double bond at the 3 position was needed to increase the activity. Moreover, the hydroxyl group at the 8-position was essential for the activity. On the time course study, thunberginols A, B and F completely inhibited histamine release by pretreatment at 100 microM for 1 to 15 min, whereas DSCG inhibited histamine release only following 1-min pretreatment at 1000 microM. These results suggested that the mechanisms of the inhibitory effect of thunberginols are different from that of DSCG. PMID- 10465419 TI - Synthesis and biological activities of phenylahistin derivatives. AB - X-ray crystallographic analysis was performed and several phenylahistin derivatives were synthesized to elucidate the structural components necessary for the anti-microtubule activity of phenylahistin. We primarily focused on the unique isoprenylated dehydrohistidine structure. Our results showed that a uniplanar pseudo-three-ring structure formed by the hydrogen bonding of diketopiperazine and imidazole rings is important for the anti-microtubule activity of phenylahistin. PMID- 10465420 TI - Design and synthesis of 3-fluoro-2-oxo-3-phenylpropionic acid derivatives as potent inhibitors of 4-hydroxyphenylpyruvate dioxygenase from pig liver. AB - Several rationally designed analogs of 3-fluoro-2-oxo-3-phenylpropionic acid were chemically synthesized, and the reactions of the hydrate form of these compounds with 4-hydroxyphenylpyruvate dioxygenase from pig liver as inhibitors were examined. Compounds 14a and 14b were found to be potent competitive inhibitors of the enzyme with Ki values of 10 and 22 microM, respectively. PMID- 10465422 TI - Three-dimensional quantitative structure activity relationships (3-D-QSAR) of antihyperglycemic agents. AB - A three-dimensional quantitative structure activity relationship study (3-D-QSAR) was performed on a set of thiazolidinedione antihyperglycemic agents using the comparative molecular field analysis (CoMFA) method. The CoMFA models were derived from a training set of 53 compounds. Fifteen compounds, which were not used in model generation were used to validate the CoMFA models. All the compounds were superimposed to the template structure by atom-based and shape based strategies. The SYBYL QSAR rigid body field fit was also used for aligning the ligands. A total of twelve different alignments were generated. The resulting models exhibited a good cross-validated r2cv values (0.624-0.764) and the conventional r2 values (0.689-0.921). A more robust cross-validation test using cross-validation by 2 groups (leave half out method) was performed 100 times to ascertain the predictiveness of the CoMFA models. The mean of r2cv values from 100 runs ranged from 0.611-0.690. Few models exhibited good external predictivity. These models were then used to define a hypothetical receptor model for antihyperglycemic agents. PMID- 10465421 TI - Synthesis and hybridization property of sugar and phosphate linkage modified oligonucleotides. AB - 1-[3-Deoxy-5-O-(4,4'-dimethoxytriphenylmethyl)-3-C-hydroxymethyl-2 -O-(2 methoxyethoxymethyl)-beta-D-erythro-pentofuranosyl]thymine (13) was synthesized from 1,2-isopropylidene-D-xylose (1) as a building block of modified oligonucleotides. Three types of novel oligonucleotides were synthesized from 13 and their T(m)s were compared with those of the corresponding natural oligonucleotides. It was found that our synthesized oligomers had lower affinity to DNA and RNA than the natural oligomers. PMID- 10465423 TI - Cortico-subcortical synchronization in the chloralose-anesthetized cat. AB - The spontaneous and paroxysmal cerebral cortical synchronized activity was used as reference to study the cortical impact exerted on subcortical neurons. The sensorimotor cortical synchronized activity spread down to subcortical structures receiving direct cortical input, including neuronal populations that originate descending rubrospinal, tectospinal and reticulospinal motor axons, and to a somatosensory relay station, the cuneate nucleus. Lesion of the pyramidal tract abolished the cortically induced synchronization of the activity of contralateral cuneate nucleus neurons. PMID- 10465424 TI - Plastic changes underlying vestibular compensation in the guinea-pig persist in isolated, in vitro whole brain preparations. AB - Vestibular compensation for the postural and oculomotor deficits induced by unilateral labyrinthectomy is a model of post-lesional plasticity in the central nervous system. Just after the removal of one labyrinth, the deafferented, ipsilateral vestibular nucleus neurons are almost silent, and the discharge of the contralateral vestibular nucleus neurons is increased. The associated static disorders disappear in a few days, as normal activity is restored in both vestibular nuclei. In this study, we searched for traces of vestibular compensation in isolated whole brains taken from adult guinea-pigs. The electrophysiological responses evoked in control brains were compared to those evoked in brains taken from animals that had previously been labyrinthectomized. Guinea-pigs compensated for an initial labyrinthectomy within three days. In vivo, subsequent deafferentation of vestibular nucleus neurons on the intact side triggered "Bechterew's phenomenon": a new postural and oculomotor syndrome appeared, similar to the one induced by the first lesion, but directed to the newly deafferented side. These disturbances would be caused by the new imbalance between the discharges of neurons in the two vestibular nuclei triggered by the second deafferentation. Experiments were designed to search for a similar imbalance in vitro in brains taken from labyrinthectomized animals, where the intact vestibular nerve is cut during the dissection. Isolated whole brains were obtained from young guinea-pigs at various times (one to seven days) following an initial labyrinthectomy. An imbalance between the resting activities of medial vestibular nucleus neurons on both sides of the brainstem was revealed in brains taken more than three days after the lesion: their discharge was higher on the compensated, initially lesioned side than on the newly deafferented side. In some cases, an oscillatory pattern of discharge, reminiscent of the spontaneous nystagmus associated in vivo with Bechterew's syndrome, appeared in both abducens nerves. These data demonstrate that most of the changes underlying vestibular compensation persist, and can thus be investigated in the isolated whole brain preparation. Brains removed only one day after the lesion displayed normal commissural responses and symmetric spinal inputs to vestibular nucleus neurons. However, an unusually large proportion of the neurons recorded on both sides of the preparation had very irregular spontaneous discharge rates. These data suggest that the first stages of vestibular compensation might be associated with transient changes in the membrane properties of vestibular nucleus neurons. Brains taken from compensated animals displayed a significant, bilateral decrease of the inhibitory commissural responses evoked in the medial vestibular nucleus by single-shock stimulation of the contralateral vestibular nerve. The sensitivity of abducens motoneurons on the initially lesioned, compensated side to synaptic activation from the contralesional vestibular nucleus neurons was also decreased. Both changes may explain the long-term, bilateral decrease of vestibular-related reflexes observed following unilateral labyrinthectomy. Spinal inputs to vestibular nucleus neurons became progressively asymmetric: their efficacy was increased on the lesioned side and decreased on the intact one. This last modification may support a functional substitution of the deficient, vestibular-related synergies involved in gaze and posture stabilization by neck related reflexes. PMID- 10465425 TI - Notch1 inhibits neurite outgrowth in postmitotic primary neurons. AB - Notch plays an important role in cell fate decisions in uncommitted proliferative cells, including neurogenesis, but is believed to not have a role in postmitotic cells. We have shown previously that Notch1 is highly expressed in embryonal mouse and human brain, but surprisingly it continues to be expressed at low levels in the adult brain. The function of Notch1 in postmitotic neurons in mammals is unknown. To better understand the potential role of Notch1 in mature central nervous system neurons we studied the effect of Notch1 transfection on neurite outgrowth in primary neocortex hippocampal neurons. Transfection at two days in vitro with full length Notch1 inhibited neurite outgrowth. Transfection at five to six days in vitro, after neurite outgrowth was established, led to apparent regression of neurites. These effects were enhanced when truncated constitutively active forms of Notch1 were introduced. Co-transfection with Numb, a physiological inhibitor of Notch, blocked Notch's effect on neurite outgrowth. We also examined whether Notch1 could activate C-promoter binding factor (CBF1) transcription factor using C-promoter binding factor-luciferase constructs, and demonstrated that this signal transduction pathway is present and can be activated in postmitotic neurons. Our results show that in postmitotic neurons Notch1 influences neurite morphology, and can activate its native signal transduction pathway. These data strongly suggest that Notch1 may play a physiologically important role in the central nervous system beyond neurogenesis. PMID- 10465426 TI - Measurement of GABAergic parameters in the prefrontal cortex in schizophrenia: focus on GABA content, GABA(A) receptor alpha-1 subunit messenger RNA and human GABA transporter-1 (HGAT-1) messenger RNA expression. AB - The hypothesis that the pathophysiology of schizophrenia may be associated with a dysfunction in GABA transmission in the human prefrontal cortex was investigated. Human post mortem brain tissue from 10 control cases and six cases of schizophrenia were processed for amino acid analysis and for radioactive in situ hybridization. Laminae III and V of three prefrontal cortical areas were examined in detail, namely Brodmann areas 9, 10 and 11. Of these three areas significant changes in GABAergic markers were found only in areas 9 and 10. Of note, a significant decrease in the tissue content of GABA was observed and this was accompanied by a marked increase in the cellular expression of the GABA(A) receptor alpha-1 subunit messenger RNA and a marked decrease in the expression of human GABA transporter-1, the messenger RNA encoding the neuronal GABA transporter protein. The amino acid analysis data provided in this study coupled with the detailed cellular study of several GABAergic markers in the human prefrontal cortex provide direct evidence in support of a disturbance in GABA transmission in the prefrontal cortex, which may be loosely termed "hypofrontality". PMID- 10465427 TI - Distribution of voltage-dependent calcium channel beta subunits in the hippocampus of patients with temporal lobe epilepsy. AB - Voltage-dependent Ca2+ channels constitute a major class of plasma membrane channels through which a significant amount of extracellular Ca2+ enters neuronal cells. Their pore-forming alpha1 subunits are associated with cytoplasmic regulatory beta subunits, which modify the distinct biophysical and pharmacological properties of the alpha1 subunits. Studies in animal models indicate altered expression of alpha1 and/or beta subunits in epilepsy. We have focused on the regulatory beta subunits and have analysed the immunoreactivity patterns of the beta1, beta2, beta3 and beta4 subunits in the hippocampus of patients with temporal lobe epilepsy (n = 18) compared to control specimens (n = 2). Temporal lobe epilepsy specimens were classified as Ammon's horn sclerosis (n = 9) or focal lesions without alteration of hippocampal cytoarchitecture (n = 9). Immunoreactivity for the beta subunits was observed in neuronal cell bodies, dendrites and neuropil. The beta1, beta2 and beta3 subunits were found mainly in cell bodies while the beta4 subunit was primarily localized to dendrites. Compared to the control specimens, epilepsy specimens of the Ammon's horn sclerosis and of the lesion group showed a similar beta subunit distribution, except for beta1 and beta2 staining in the Ammon's horn sclerosis group: in the severely sclerotic hippocampal subfields of these specimens, beta1 and beta2 immunoreactivity was enhanced in some of the remaining neuronal cell bodies and, in addition, strongly marked dendrites. Thus, hippocampal neurons apparently express multiple classes of beta subunits which segregate into particular subcellular domains. In addition, the enhancement of beta1 and beta2 immunoreactivity in neuronal cell bodies and the additional shift of the beta1 and beta2 subunits into the dendritic compartment in severely sclerotic hippocampal regions indicate specific changes in Ammon's horn sclerosis. Altered expression of these beta subunits may lead to increased currents carried by voltage-dependent calcium channels and to enhanced synaptic excitability. PMID- 10465428 TI - Alterations of perisomatic GABA synapses on hippocampal CA1 inhibitory interneurons and pyramidal cells in the kainate model of epilepsy. AB - In the kainate model of epilepsy, electrophysiological and anatomical modifications occur in inhibitory circuits of the CA1 region of the rat hippocampus. Using postembedding GABA immunocytochemistry and electron microscopy, we characterized perisomatic GABA and non-GABA synaptic contacts in CA pyramidal cells, and GABAergic interneurons of stratum oriens/alveus and stratum lacunosum-moleculare, and examined if changes occurred at these synapses at two weeks post-kainate treatment. We found that, in control rats, the number and total length of perisomatic GABA synapses were significantly smaller (approximately 40-50%) in lacunosum-moleculare interneurons than in oriens/alveus interneurons and pyramidal cells. Additionally, the number and total length of perisomatic non-GABA synapses were different among all cell types, with these parameters increasing significantly in the following order: pyramidal cells3 cm prior to surgery (n = 24) or metastatic disease (n = 15) received cyclophosphamide 600 mg/m2 and doxorubicin 50 mg/m2 as an intravenous bolus every 3 weeks for six courses. Continuous infusional 5-FU was delivered via a central venous line for a maximum of 18 weeks at dose levels of 100 mg/m2 per day (n = 6), 150 mg/m2 per day (n = 3) and 200 mg/m2 per day (n = 30). At the 200 mg/m2 per day dose level, 36% of patients required dose delays and 23% dose reductions; there was one death due to neutropenic sepsis. Hickman line complications occurred at all dose levels, particularly thrombosis (18%) and infection (33%). The response rate was 62% (95% confidence interval (CI) 32-84) for metastatic disease, including five complete responses (CRs). The response rate for primary tumours prior to surgery was 81% (95% CI 57-95) including six clinical CRs. Infusional FAC is an active regimen and has an acceptable toxicity profile. It does not, however, appear to offer any significant advantage over other chemotherapy regimens. This study does not support the further evaluation of infusional 5-FU at these doses in combination with doxorubicin and cyclophosphamide. PMID- 10465472 TI - Treatment of endotracheal or endobronchial obstruction by non-small cell lung cancer: lack of patients in an MRC randomized trial leaves key questions unanswered. Medical Research Council Lung Cancer Working Party. AB - Symptoms of endotracheal or endobronchial obstruction caused by non-small cell lung cancer (NSCLC) may be relieved with external beam radiotherapy (XRT) or endobronchial treatment. The comparative roles of these two methods need to be established. Patients with predominantly intraluminal obstruction of the trachea, a main bronchus or a lobar bronchus by unresectable NSCLC were randomized to XRT versus the clinician's choice of endobronchial treatment with brachytherapy, laser resection or cryotherapy, according to local availability and practice. Clinicians' assessments included symptoms of obstruction, WHO performance status, lung function tests and adverse effects of treatment. Patients completed a Rotterdam Symptom Checklist at all assessments and a daily diary card to record the severity of major symptoms during the first 4 weeks. To show a difference of 15% in the relief of breathlessness rates at 4 months (from 65% to 80%), 400 patients were required. In spite of our many previously successful lung cancer trials, and initial interest from clinicians in 24 UK centres, who estimated they could randomize 200 patients per year into the present trial, only 75 patients were randomized from seven centres over 3.5 years. Intake to the trial was therefore abandoned in November 1996 although an independent Data Monitoring and Ethics Committee had concluded in April 1996 that the scientific case for the trial was still strong; there were no competing trials; there were no design problems; and much had been done to promote the trial. The main reasons given by centres for the slow intake were: lack of referrals of untreated patients; patients being referred specifically for endobronchial treatment; patients having already received XRT; emergency endobronchial relief of obstruction being necessary; and XRT and endobronchial treatment being considered complementary and not as alternatives. The relative advantages and disadvantages of XRT versus endobronchial treatment remain to be determined. The lack of recruitment to this trial raises the issue of innovative techniques not being given the chance of proving their worth compared with traditional treatments. PMID- 10465473 TI - Interstitial pneumonitis following mitozantrone, chlorambucil and prednisolone (MCP) chemotherapy. AB - We describe two cases of interstitial lung disease in patients with non-Hodgkin's lymphoma who were treated with combination chemotherapy including mitozantrone. In both we had radiological and histological evidence of interstitial lung disease, with patterns of organizing pneumonia and hypersensitivity without a clear aetiological agent. Clinical resolution occurred on withdrawal of chemotherapy. One patient required a course of corticosteroid treatment. To date, both patients are well and in remission, and there has been no recurrence of their respiratory disease. We postulate that these observations are hitherto undescribed pulmonary abnormalities secondary to mitozantrone therapy. PMID- 10465474 TI - Myelodysplastic syndrome as a complication of neo-adjuvant triple M chemotherapy and radiotherapy. AB - Cases of myelodysplastic syndrome occurring after multi-drug chemotherapy are rare; they are more often associated with the use of alkylating agents. We report the case history of a patient with myelodyspasia occurring after neoadjuvant methotrexate, mitoxantrone and mitomycin C (triple M) chemotherapy with subsequent radiotherapy for locally advanced breast cancer. Cytogenetic analysis of a bone marrow biopsy confirmed the typical chromosomal abnormalities associated with therapy related myelodysplasia. Few treatments for this disorder have been found to be of value. The aetiology, incidence and management options are briefly discussed. PMID- 10465475 TI - Splenosis mimicking metastases from breast carcinoma. AB - The case history is presented of a patient with breast cancer who was extensively investigated for presumed hepatic metastases, which were finally diagnosed as splenosis, the heterotopic autotransplantation of splenic tissue after traumatic rupture of the spleen. This case history highlights the importance of obtaining a pathological diagnosis prior to labelling a patient as having metastatic disease. This is especially important for patients who have an unusual pattern or appearance of metastases or for whom the risk of metastatic disease is presumed to be minimal. PMID- 10465476 TI - Life-threatening tamoxifen-induced hypercalcaemia. AB - This case report serves to emphasize two important features of metastatic breast carcinoma. First, that tamoxifen-induced flare, although a rare and self-limiting phenomenon, may be fatal and must thus be recognized and treated promptly. Secondly, those patients presenting with hypercalcaemia, as part of tamoxifen induced tumour flare, invariably have metastatic disease but they may enjoy a durable prognosis if this is confined to the skeleton. PMID- 10465477 TI - Subcutaneous calcification as a late effect of orthovoltage chest wall irradiation. AB - Adjuvant radiotherapy to the breast or chest wall is given to some patients with breast cancer, to reduce the risk of local recurrence. It is known to be associated with various late sequelae, including subcutaneous fibrosis, telangiectasia and pulmonary fibrosis. Delivering radiotherapy to the chest wall and nodal drainage areas presents the technical problem of matching the glancing and anterior supraclavicular fields. Overlap between these fields will result in underlying tissues receiving a larger dose than intended; similarly, a gap results in an inadequate dose. We present the case history of a patient with subcutaneous calcification occurring as a late sequela of radiotherapy to the chest wall and anterior supraclavicular field. This has not been previously reported and is thought to have arisen from a high dose region in an area of overlap between fields. PMID- 10465478 TI - Hyperbaric oxygen corrects sacral plexopathy due to osteoradionecrosis appearing 15 years after pelvic irradiation. AB - In 1982, a 55-year-old woman was treated by total cystectomy and adjuvant radiotherapy/chemotherapy for a leiomyosarcoma of the bladder. Fifteen years later she presented with symptoms and signs of sacral plexopathy. Investigations revealed osteoradionecrosis of the sacrum. Hyperbaric oxygen therapy (HBO2) was instituted and progressive resolution of the neurological complaints followed. HBO2 should be considered when managing late-onset sequelae in previously irradiated patients. PMID- 10465479 TI - Bilateral renal cancer in children: a difficult, challenging and changing management problem. AB - Bilateral disease occurs in 6% of patients with Wilms' tumour. Bilateral renal involvement is present in 25%-50% of children with B-cell non-Hodgkin's lymphoma. Paediatric oncologists therefore encounter bilateral renal disease relatively frequently. A patient with bilateral Wilms' tumour is presented, in whom at least 16 synchronous tumours in the kidneys were treated successfully by primary chemotherapy and 'nephron-sparing' surgery, without renal radiotherapy. We believe the successful treatment without radiotherapy will allow greater potential for normal growth in the future. The case history of a patient with renal failure due to the infiltration of both kidneys by lymphoma and who was treated successfully by chemotherapy, is also presented. One kidney has completely returned to normal function and growth; the other has completely failed and almost disappeared, demonstrating complete reversibility of the damage up to some critical point, beyond which, failure and atrophy result. These two case histories demonstrate extreme examples of clinical dilemmas involving bilateral renal tumours and allow discussion of modern management aimed at preserving renal function. PMID- 10465480 TI - Systemic metastases of glioblastoma multiforme. AB - The case history of a 40-year-old man who developed systemic metastases 2 years after treatment for glioblastoma is reported. The diagnosis was confirmed by biopsy. The role of immunohistochemistry in the diagnosis of this rare event is discussed and illustrated. PMID- 10465481 TI - Complete resolution of angiosarcoma of the scalp with liposomal daunorubicin and radiotherapy. AB - Angiosarcoma of the head and neck is a rare malignant vascular tumour that predominantly affects elderly men and generally has a poor prognosis. Complete responses to treatment for larger tumours are uncommon. A patient with an extensive tumour that showed a complete response to combination therapy with liposomal daunorubicin and radiotherapy is presented. The rationale for using liposomal chemotherapy agents in the treatment of vascular tumours such as angiosarcoma is discussed. PMID- 10465482 TI - Adolescents and young adults with cancer: towards a seamless service. PMID- 10465483 TI - Radiotherapy for age-related macular degeneration. PMID- 10465484 TI - Patient empowerment: individual control of health benefits. PMID- 10465485 TI - Medium- and long-term results of high tibial osteotomy for varus gonarthrosis in an agricultural population. AB - High tibial valgus osteotomy for varus gonarthrosis was performed in 63 consecutive patients in a homogenous agricultural population using two different surgical techniques. Patients were divided into two groups. A two-level Mittelmeier osteotomy was performed in group A patients, and a lateral closed wedge high tibial osteotomy using the AO/ASIF L-plate was performed in group B patients. Operations were performed by two different groups of surgeons. Patients were evaluated postoperatively for correction of knee axis, functional result, subjective impression, and complications. In group A patients, 80% of the operated knees were corrected to the mechanical axis and in group B patients, 82% of the knees were corrected to 6 degrees-10 degrees valgus of the anatomical axis. Ninety percent, 70%, and 54% of group A and 91%, 73%, and 57% of group B patients were rated as satisfactory results at 5, 9, and 12 years postoperatively, respectively; these differences were not statistically significant. One year postoperatively, 91% of group A and 96% of group B patients reported their symptoms had improved. However, patient satisfaction decreased at 5, 7, and 12 years postoperatively, with 91%, 89%, and 66% of group A and 96%, 93%, and 68%, respectively, of group B patients reporting their symptoms had improved; these differences were not statistically significant. Postoperatively, most patients returned to full agricultural activity. Total knee arthroplasty, which was later required in 12% of the knees, was not significantly jeopardized by the previous osteotomy. PMID- 10465486 TI - Treatment of large skeletal defects in the lower extremities using double-strut, free vascularized fibular bone grafting. AB - This article reports on the use of double-strut, free vascularized fibular grafts to treat six patients with infected nonunion or traumatic bone loss in the femur or tibia after prolonged treatment and multiple operations. The defects were 6-13 cm long. Five patients achieved solid union within 6 months, and one patient required additional cancellous grafting to achieve union at the distal end of the fibula. One patient experienced a stress fracture due to strenuous exercise, and union was achieved 3 months after reapplying an external fixator. Although three patients had some restricted knee motion, all patients had a satisfactory outcome in regard to walking, and no limb-length discrepancies were noted in any patient. PMID- 10465487 TI - Revision total hip arthroplasty in patients with avascular necrosis. AB - This retrospective study compared the results of revision total hip arthroplasty (THA) in patients with avascular necrosis to patients with osteoarthritis. Twenty two revision THAs (34 components) in 19 patients with avascular necrosis were compared with 35 revision THAs (55 components) in 31 patients with osteoarthritis. All of the procedures were performed by a single surgeon between 1981 and 1994, and all patients had a minimum of 2 years of follow-up. At the time of revision surgery, the average age was 54 years for the avascular necrosis patients and 67 years for the osteoarthritis patients (P=.002). Clinical and radiographic follow-up was performed for an average of 7 years (range: 2-12 years). Six (18%) components in the avascular necrosis group and nine (16%) components in the osteoarthritis group required re-revision for aseptic loosening an average of 7 years after the original revision (range: 2-11 years). Statistical analysis demonstrated no significant difference between the two groups in regard to the incidence of failure, time to failure, Harris Hip Score, and radiographic appearance. Contrary to expectations derived from the literature on primary THA, no difference was found between the outcome and survivorship of revision THA in patients with avascular necrosis and patients with osteoarthritis, despite a significant difference in age. PMID- 10465488 TI - Management of vertebral diskitis and osteomyelitis. PMID- 10465489 TI - "Tug-of-war-hand": transforearm amputation by an unusual mechanism. PMID- 10465490 TI - Piriformis syndrome resulting from an anomalous relationship between the sciatic nerve and piriformis muscle. PMID- 10465491 TI - Isolated granulocytic sarcoma of the humerus. PMID- 10465492 TI - Radiologic case study. Diffuse red bone marrow hyperplasia of the spine, pelvis and femurs. PMID- 10465493 TI - Focal extratemporal epilepsy: clinical features, EEG patterns, and surgical approach. AB - The objective of this review is a summary of the clinical and electrographic findings in those forms of epilepsy to which the term 'extratemporal' (ExT) can be applied. They form a group that differs in many ways from the better known temporal lobe epilepsies. Seizure foci are difficult to localize by clinical semiology alone but modern imaging now often allows a precise definition of the epileptogenic area. The most common causes of ExT epilepsy are tumors and cortical dysgenesis. The concept of 'dual pathology' implies the coexistence of two or more distinct lesions, typically mesial temporal sclerosis and cortical dysplasia. Electroencephalography (EEG) and electrocorticography (ECoG) are valuable tests in the definition of the epileptogenic area beyond the structural lesion, and surgical removal must be guided by the nature of the lesion and the extent of the epileptogenic zone. PMID- 10465494 TI - Proton MR spectroscopy in clinically isolated syndromes suggestive of multiple sclerosis. AB - The concentration of the metabolite N-acetyl aspartate (NAA), thought to be a marker of axonal loss or damage, has been shown to be reduced in lesions, as demonstrated by high signal areas on T2-weighted MRI, and in normal-appearing white matter (NAWM) in established multiple sclerosis (MS). The stage of the disease when these changes first appear is not known. To try to determine this we studied 20 patients with clinically isolated syndromes, many of whom will be at the earliest clinical stages of MS, and 20 age- and sex-matched controls with single-voxel proton magnetic spectroscopy (MRS). MRS was performed using a General Electric 1.5T Signa EchoSpeed scanner (TR 3000 ms, TE 30 ms, PRESS). Absolute metabolite concentrations were determined using the LCModel fitting software. No significant reduction of NAA concentration was evident in the NAWM of the patients (patients: median 7.3 mM; controls: median 7.7 mM; P=0.19). There was, however, a significantly lower concentration of NAA in lesions (median 6.6 mM, P=0.015). Absolute values of choline-containing compounds, creatine and myo inositol were significantly raised in the lesions (P=0.007, P=0.011 and P=0.002 respectively). The low NAA in lesions is consistent with axonal loss, damage or dysfunction occurring focally at the earliest clinical phase of the disease. The lack of any significant reduction in NAA in patient NAWM demonstrates that more widespread axonal changes are not yet detectable at this early clinical stage. A larger cohort and follow-up will be necessary to determine whether or not MRS findings have any prognostic significance for individual patients or sub-groups. This will also enable the clarification of the time course, pathogenesis and pathophysiological significance of the development of the low NAA, which is found in the NAWM of many patients with established MS. PMID- 10465495 TI - Increased frequency of recognition of delipidated versus intact CNS myelin in multiple sclerosis and control subjects. AB - The proliferative response of mononuclear cells from MS patients and normal control subjects to intact and delipidated myelin membranes was examined. The mean frequency of recognition in both groups of human subjects was greater for delipidated myelin than for intact myelin. Human T cell lines established using intact or delipidated myelin as the antigen were highly heterogeneous in response, and were each able to recognize myelin basic protein and myelin proteolipid protein peptides. However, there was no difference in the frequency of recognition of either form of myelin membrane when MS patients were compared to control subjects. Our results suggest that the presentation of delipidated forms of membrane proteins might enhance the response to myelin antigens in vivo, and be relevant to demyelinating diseases. PMID- 10465496 TI - Guamanian neurodegenerative disease: electrophysiologic findings. AB - Amyotrophic lateral sclerosis (ALS), parkinsonism and/or dementia are highly prevalent among the Chamorro population of Guam. The incidence of Guamanian ALS has markedly declined in recent years, but these incidence figures may reflect underascertainment of subclinical disease. Guamanian Chamorro patients have not been systematically studied using modern clinical neurophysiological techniques. Electromyography (EMG: needle exam and nerve conduction studies) was used to study 29 patients with the major subtypes of Guamanian neurodegenerative disease, as well as 11 neurologically normal Guamanian Chamorro subjects. Central conduction was assessed by somatosensory evoked potentials (SEP's) in 16 patients. EMG evidence of peripheral neuropathy, (often subclinical) was found in 45% of Guamanian patients but no Chamorro control subjects. Diabetes mellitus, which is highly prevalent in this population, was present in some, but not all of these cases. Clinically unsuspected motor neuron disease was identified by EMG in only one of the 23 Guamanian patients with parkinsonism and/or dementia and in none of the 11 Chamorro control subjects. Two of seven patients with the clinical phenotype of Guamanian ALS had a more benign EMG pattern on the needle electrode exam with absence of fibrillation and fasciculation potentials. Three of 16 patients (all with parkinsonism and dementia) had mildly abnormal tibial SEP's. No patient had EMG evidence of myopathy or a defect of neuromuscular transmission. We conclude: (1) peripheral neuropathy may be a manifestation of Guamanian neurodegenerative disease; (2) the declining prevalence of ALS on Guam is not associated with the development of a subclinical form of motor neuron disease; (3) the substantial overlap of Guamanian ALS with parkinsonism-dementia reported in prior decades is no longer apparent; (4) abnormal central conduction, as assessed by tibial SEP's, is present in some patients with Guamanian parkinsonism-dementia. PMID- 10465497 TI - A case of McLeod syndrome with unusually severe myopathy. AB - A 51-year-old man developed weakness and muscle atrophy in the legs at the age of 41, later followed by choreiform involuntary movements. Neurological and laboratory examinations revealed severe muscle weakness and atrophy, and areflexia in all the extremities, acanthocytosis and an elevated serum creatine kinase level. Together with these findings, the weak expression of Kell blood group antigens and the absence of the Kx antigen led to a definite diagnosis of McLeod syndrome for his condition. Brain magnetic resonance imaging revealed marked atrophy of the head of the caudate nuclei. Although immunocytochemical analysis of dystrophin in muscle specimens from our patient revealed normal staining, we found prominent fiber size variability, central nuclei, and connective tissue proliferation as well as necrotic and regenerating fibers, which are as a whole compatible with the myopathology of muscular dystrophy. Moreover, muscle computerized tomography of the lower extremities revealed the 'selectivity pattern' characteristically reported in muscular dystrophies including Duchenne type muscular dystrophy. The muscular symptoms and pathology in McLeod syndrome have been reported to be mild, but the present case clearly shows that the muscular features in this condition may be much more severe than previously thought. PMID- 10465498 TI - Increased peripheral nerve microvessels in early experimental diabetic neuropathy: quantitative studies of nerve and dorsal root ganglia. AB - Microangiopathy is an important complication of diabetes mellitus and neovascularity is a feature of human diabetic retinopathy. The objective of this work was to evaluate numbers, areas and size distributions of whole nerve, endoneurial and dorsal root ganglia perfused microvessels in a detailed fashion using unfixed tissues from rats with experimental diabetes. Experimental neuropathy was studied in male Sprague-Dawley rats 12 weeks after streptozotocin or citrate buffer injection. Electrophysiological recordings of sciatic-tibial motor and caudal sensory fibers identified conduction slowing in diabetes indicating neuropathy. Diabetics had a rise in the numbers of whole nerve microvessels and endoneurial microvessels with associated rises in vessel densities and total vessel luminal areas. Increased vessel numbers in 15-30 microm diameter size ranges were particularly prominent. There was a rise in summed vascular areas in diabetics but the mean luminal area of vessels was not increased. Similar, but not significant trends were observed in a selective analysis of endoneurial vessels alone. In contrast, dorsal root ganglia microvessels were not increased in number. Early experimental diabetic neuropathy is associated with increased numbers of microvessels supplying the peripheral nerve trunk, likely representing neovascularity. PMID- 10465499 TI - Association of vitamin D receptor gene polymorphism with multiple sclerosis in Japanese. AB - 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3), the biologically active form of vitamin D, exerts an immunosuppressive effect and can completely prevent experimental autoimmune encephalomyelitis (EAE). 1,25(OH)2D3 exerts most of its actions only after it has bound to its specific nuclear receptors. To investigate the possible role of vitamin D receptor gene (VDRG) polymorphism in susceptibility to or disease-modulation of MS, we evaluated 77 Japanese patients with 'conventional' MS and 95 controls. A VDRG allelic polymorphism was assessed by Bsm1 endonuclease restriction after specific PCR amplification. Genotypic polymorphism was clearly defined as BB (absence of restriction site on both alleles), bb (presence of restriction site on both alleles), or Bb (heterozygous). We found overexpression of the b allele (92.9 vs. 84.2%: P=0.0138) and homozygote bb (85.7 vs. 71.6%; P=0.0263) in MS patients compared with controls. The results indicate for the first time an association of MS with VDRG polymorphism, which may be involved in pathogenesis of MS, or in the linkage disequilibrium of VDRG to another pathogenic gene loci. The role of VDR gene polymorphism should be further studied in other populations, and the distribution of other polymorphism, such as Apa I, Taq I, should be also analyzed to confirm another susceptibility gene for MS and to obtain more adequate strategies for treatment of MS. PMID- 10465500 TI - Changes in the clinical phenotypes of multiple sclerosis during the past 50 years in Japan. AB - In order to clarify whether or not marked changes in the social environment during the past 50 years in Japan may have altered the clinical phenotypes of multiple sclerosis (MS), we retrospectively analyzed 143 consecutive patients with clinically definite MS who developed the disease between 1950 and 1997. Fifty-two patients were classified as Asian type MS showing a selective involvement of the optic nerves and the spinal cord, while 91 patients were considered to have Western type MS which demonstrated the involvement of multiple sites in the central nervous system including the cerebrum, cerebellum and brainstem. The ratio of Asian type versus Western type MS was 1:0.5 in the patients born in the 1920s and 1:1.27, 1:1.64 and 1:1.7 in those born in the 1930s, 1940s and 1950s, respectively, and thereafter it increased to 1:4.67 in those born in the 1960s and 1:4 in those born after the 1970s. As a result, the proportion of Asian type MS significantly decreased in the patients born after 1960 as compared with those born from 1930 to 1959 (P=0.0121). In the Asian type MS, the age of onset was significantly higher in the patients who developed the disease from 1985 to 1997 (42.4+/-13.5 years) than in those who developed the disease from 1950 to 1984 (32.3+/-12.4 years) (P=0.0149), while in the Western type MS no such change in the age of onset was observed. These findings suggest that the frequency of Asian type MS has apparently decreased in younger Japanese born after 1960 when Japan's rapid economic growth had just started, and environmental factors are therefore considered to contribute to determine the clinical phenotypes of MS in Asians. PMID- 10465501 TI - Brain MRI correlates of magnetization transfer imaging metrics in patients with multiple sclerosis. AB - Previous studies suggested that magnetization transfer ratio (MTR) histograms are highly correlated with other magnetic resonance imaging (MRI) measures and can be used as a reliable method for quantifying overall disease burden in multiple sclerosis (MS). However, the relative influence of burden and severity of macroscopic MS lesions and degree of brain atrophy on various MTR histogram parameters has not yet been fully elucidated. Aim of the present study was to investigate which MRI measure best predicts the values of MTR histogram parameters in MS patients. Forty-two MS patients underwent brain dual-echo. T1 weighted and magnetization transfer imaging (MTI) MRI scans. Hyperintense lesion load (LL) on proton density (PD)-weighted and hypointense LL on T1-weighted images were measured using a local thresholding technique. A measure of brain atrophy was derived from T1-weighted images by computing the volume of brain tissue segmented from a slab of five consecutive slices rostral to the velum interpositum. On MTI scans, MTR histogram analysis was performed for the whole brain and average lesion MTR was also calculated. PD-weighted LL, T1-weighted LL and brain volume were significantly correlated with several MTI-derived measures. When a multivariate analysis was performed, brain volume alone significantly predicted the values of all the MTR histogram-derived measures (P values ranged from 0.003 to 0.0002). The ratio between hypointense T1-weighted and hyperintense PD-weighted LL significantly predicted average lesion MTR (P<0.05). Our results confirm that MTR can be used as a reliable method to assess both the overall disease burden and the individual lesion intrinsic nature in MS patients. The significant influence of brain atrophy on MTR histogram parameters supports the concept that this method also provides information on the loss of brain parenchyma in MS. PMID- 10465502 TI - Disconnection of cerebellar Purkinje cells in Kearns-Sayre syndrome. AB - Kearns-Sayre syndrome (KSS) is a sporadic multisystem disorder due to rearrangements in mitochondrial DNA (mtDNA). To gain further insight into the pathogenesis of cerebellar dysfunction in KSS, antibodies against synaptophysin (SY) were used to identify presynaptic terminals and antibodies to calbindin D (CB) to identify Purkinje cells in the cerebellar cortex and in the dentate nucleus from two autopsied cases of KSS. By conventional neuropathology we found marked spongiform degeneration and by immunohistochemistry a disruption of presynaptic terminals and of the terminal arborizations of Purkinje cell axons on multipolar neurons of the dentate nucleus in the KSS patients. We suggest that a disconnection of Purkinje cells at the dentate nucleus may play a role in the pathogenesis of cerebellar ataxia in KSS. PMID- 10465503 TI - Variation in the number of CAG repeats in the Machado-Joseph disease gene (MJD1) in the Japanese population. AB - Variation in the number of CAG repeats in the Machado-Joseph disease gene (MJD1) was examined by polymerase chain reaction and denaturing polyacrylamide gel electrophoresis analysis of 2134 normal and 135 affected chromosomes of Japanese individuals. The number of repeats ranged from 14 to 47 in normal alleles and from 61 to 84 in disease-associated alleles. The most frequent and lowest number of repeats was 14. The size distribution of normal MJD1 alleles did not fit a normal distribution curve, but was tetramodal. Repeats from 14 to 17, 18 to 23, 24 to 25, and 26 to 47 units were designated groups A through D, respectively. When examined Hardy-Weinberg equilibrium by chi-square analysis of goodness of fit: no evidence of significant deviation from the Hardy-Weinberg equilibrium was observed [x2=4.248<16.919 (P=0.05), df=9]. The observed distribution peak of normal MJD1 alleles corresponding to peptides containing 10, 15, 20, and 24 glutamine suggests that stretches of 5 and 10 glutamine might constitute a functional domain of human MJD1. PMID- 10465504 TI - Peptidergic innervation of the human clitoris. AB - In the present study, the distributions of neuropeptides in the normal human clitoris and in a clitoris from an adrenogenital syndrome (AGS) was demonstrated by immunohistochemistry (IHC). Immunohistochemical screening detected a complex network of nerve fibers containing vasoactive intestinal polypeptide (VIP), peptide histidine methionine (PHM), neuropeptide tyrosine (neuropeptide Y), C flanking peptide of neuropeptide Y (CPON), calcitonin gene-related peptide (CGRP) and substance P immunoreactivities. Special attention was given to the VIP related peptide helospectin, that has been detected in neuronal elements in the clitoris. No visible differences between the localization and distribution of peptidergic nerve fibers of normal and hypertrophic clitoris from AGS have been observed. Co-localization studies showed the co-existence of VIP, PHM and partly helospectin and neuropeptide Y with CPON within nerve fibers in the cavernous tissue and substance P and CGRP co-expression in nerve fibers especially underneath and within the glans clitoris. PMID- 10465505 TI - Effects of oral fructose and glucose on plasma GLP-1 and appetite in normal subjects. AB - Oral glucose is a potent stimulant of glucagon-like peptide-1 (GLP-1) secretion. The effect of oral fructose on GLP-1 secretion in humans is unknown. The aims of this study were to determine (i) whether oral fructose stimulates GLP-1 secretion and (ii) the comparative effects of oral glucose and fructose on appetite. On 3 separate days, 8 fasting healthy males received, in single-blind randomized order (i) 75 g glucose, (ii) 75 fructose, or (iii) 75 g glucose followed by 75 g fructose I h later. Venous glucose, insulin and GLP-1 were measured. Appetite was assessed by visual analog questionnaires and intake of a buffet meal. Whereas glucose and fructose both increased plasma glucose, insulin and GLP-1 (P < 0.000)] for all), the response to glucose was much greater (P < 0.005 for all). There was no increase in plasma GLP-1 when fructose was given after glucose. There was no difference in food intake after oral glucose or fructose. We conclude that oral fructose (75 g) stimulates GLP-1 (and insulin) secretion, but the response is less than that to 75 g glucose. These observations suggest that neither GLP-1 nor insulin play a major role in the regulation of satiation. PMID- 10465506 TI - 17beta-estradiol regulation of melanin-concentrating hormone and neuropeptide-E-I contents in cynomolgus monkeys: a preliminary study. AB - Melanin-concentrating hormone (MCH) and neuropeptide-E-I (NEI) regulate several behaviors and neuroendocrine functions in rats. Possible influence of these peptides on sexual behavior and reproduction in mammals other than rodents prompted us to investigate: 1) The sites of synthesis of MCH and NEI in the brain of a non-human primate (M. fascicularis); 2) The effect of 17 beta-estradiol (E2) benzoate (E2B) on pro-MCH-derived peptide concentrations in the hypothalamus of the ovariectomized (OVX) cynomolgus monkeys (M. fascicularis). Expression of MCH mRNA and peptides was examined by Northern blotting, RT-PCR and RP-HPLC/RIA. Our results demonstrate that the MCH gene is predominantly expressed in hypothalamus of macaque. E2B exposure of OVX monkeys provoked parallel phasic variations in the MCH-immunoreactivity (IR) and NEI-IR. NEI-IR and to a lesser extent MCH-IR, showed a transient increase (associated with the estradiol peak) at 30 h with a final rise of both MCH-IR and NEI-IR observed at the time (72 h post E2B) of the luteinizing hormone (LH) surge. RP-HPLC analysis of peptide extracts revealed the presence, in addition to mature MCH and NEI, of different MCH-IR and NEI-IR forms in the hypothalami of control and E2B-treated monkeys. Taken together, our results indicated that hypothalamic MCH and NEI contents are regulated after E2B treatment and they suggest the possible involvement of these peptides in the regulation of the pre-ovulatory midcycle LH surge in primates. PMID- 10465507 TI - Isolation and structure determination of a paralytic peptide from the hemolymph of the silkworm, Bombyx mori. AB - A peptide with paralytic activity in larvae of the silkworm, Bombyx mori, was isolated from its hemolymph. Purification procedures consisted of extraction with 50% acetone, Vydac C4 reversed-phase cartridge elution and 4 steps of reversed phase HPLC. Injection of the purified peptide into 4th instar B. mori larvae caused rapid and rigid paralysis for 2 min at a dose of 3.4 ng/larva. This paralytic peptide consists of 23 amino acid residues containing 2 cysteines with an intra-disulfide bond. The complete amino acid sequence is: H-Glu-AsnPhe-Val Gly-Gly-Cys-Ala-Thr-Gly-Phe-Lys-Arg-Thr-Ala-Asp-G ly-Arg-Cys-Lys-Pro-Thr-Phe-OH. The relationship between structure and the biologic activity of synthetic analogs indicated that the entire amino acid sequence and the intra-disulfide bond were necessary for biological activity. PMID- 10465508 TI - Gastrin- and cholecystokinin-like immunoreactivities in the nervous system of the earthworm. AB - The distribution of cholecystokinin and gastrin-like immunoreactive cell bodies and fibers in the nervous system of 2 annelid worms, Lumbricus terrestris and Eisenia fetida, has been studied by means of immunohistochemistry. The cerebral ganglion contains 170-250, the subesophageal ganglion contains 120-150, and the ventral ganglia contain 50-75 cholecystokinin immunoreactive cells, that represent 8-12%, 8-10% and 4-5% of the total cell number, respectively. The anti gastrin serum stained 330-360 nerve cells in the cerebral, 32-46 in the subesophageal and 7-25 in the ventral cord ganglia, representing 15-16%, 2-3% and 0.5-2% of the total cell number. Immunopositivity was found with both antisera in the enteric nervous system, where the stomatogastric ganglia and the enteric plexus contain immunoreactive cells and fibers. Immunopositive cells were found in the epithelial and subepithelial cells, as well as in nerve cells innervating the muscular layer of the gastrointestinal tube. Various epidermal sensory cells also displayed strong immunoreactivity. According to our findings and the results of several functional studies, it is suggested that in annelids cholecystokinin- and gastrin-like peptides may be involved in digestive regulation, sensory processes and central integrating processes. PMID- 10465509 TI - Cytosolic Ca2+ oscillations by gastrin releasing peptide in single HIT-T15 cells. AB - In single, superfused, FURA-2AM loaded insulin producing HIT-T15 cells, gastrin releasing peptide (GRP) induced a peak in cytoplasmnic Cu2+ ([Ca2+]i) followed by a sustained (high GRP concentrations) or oscillatory (low GRP concentrations) [Ca2+]i pattern. The GRP (25-50 microM)-induced [Ca2+]i oscillations ceased upon removal of glucose or addition of thapsigargin (1 microM), EGTA (2 mM), or diazoxide (200 microM), whereas nifedipine (10 microM) reduced their amplitude (by 35%). Both protein kinase C (PKC)-activation or PKC-inhibition disrupted GRP induced [Ca2+]i oscillations. GRP induced [Ca2+]i oscillations in insulin producing cells therefore rely on intracellular Ca2+ mobilization, voltage dependent and voltage-independent Ca2+ entry mechanisms and the integrity of protein kinase C. PMID- 10465511 TI - Leptin and neuropeptide Y (NPY) modulate nitric oxide synthase: further evidence for a role of nitric oxide in feeding. AB - Recent studies have suggested a role for nitric oxide in the regulation of food intake. Neuropeptide Y (NPY) is one of the most potent orexigenic agents. Chronic administration of leptin decreases food intake. This study examined the effects of NPY and leptin on nitric oxide synthase (NOS) in the hypothalamus. Previously it has been demonstrated that obese (ob/ob) mice have elevated NOS levels in the hypothalamus. In this study we demonstrated that the administration of leptin (6 microg/day) subcutaneously (SC) for 3 days decreased body weight (P < 0.001) and food intake P < 0.001) in obese (ob/ob) mice as expected. In addition, leptin decreased NOS in the hypothalamus nu 37% (P < 0.01) and in brown adipose tissue by 69% (P < 0.01) but not in white adipose tissue. NPY was administered intracerebroventricularly to CD-1 mice at doses of 0.25 and 0.50 microg. Mice were sacrificed 15 min after injection and NOS was measured in their hypothalami. NPY at the lower dose increased NOS in the hypothalamus by 147%. These results, taken together, with previously published studies support the concept that nitric oxide may play a role as a mediator of the effects of NPY and leptin on food intake. The alterations of NOS in brown adipose tissue following leptin administration could result in changes in blood flow or metabolism in the brown fat. PMID- 10465510 TI - Gastric inhibitory polypeptide stimulates glucocorticoid secretion in rats, acting through specific receptors coupled with the adenylate cyclase-dependent signaling pathway. AB - Gastric inhibitory polypeptide (GIP) is a 42-amino acid peptide, belonging to the VIP-secretin-glucagon superfamily, some members of this group are able to regulate adrenocortical function. GIP-receptor mRNA has been detected in the rat adrenal cortex, but investigations on the effect of GIP on steroid-hormone secretion in this species are lacking. Hence, we have investigated the distribution of GIP binding sites in the rat adrenal gland and the effect of their activation in vivo and in vitro. Autoradiography evidenced abundant [125I]GIP binding sites exclusively in the inner adrenocortical layers, and the computer-assisted densitometric analysis of autoradiograms demonstrated that binding was displaced by cold GIP, but not by either ACTH or the selective ACTH receptor antagonist corticotropin-inhibiting peptide (CIP). The intraperitoneal (IP) injection of GIP dose-dependently raised corticosterone, but not aldosterone plasma concentration: the maximal effective dose (10 nmol/rat) elicited a twofold increase. GIP did not affect aldosterone and cyclic-AMP release by dispersed zona glomerulosa cells. In contrast, GIP enhanced basal corticosterone secretion and cyclic-AMP release by dispersed inner adrenocortical cells in a concentration dependent manner, and the maximal effective concentration (10(-7) M) evoked 1.5- and 2.4-fold rises in corticosterone and cyclic-AMP production, respectively. GIP (10(-7) M) did not display any additive or potentiating effect on corticosterone and cyclic-AMP responses to submaximal or maximal effective concentrations of ACTH. The corticosterone secretagogue action of 10(-7) M GIP was abolished by the protein kinase A (PKA) inhibitor H-89 (10(-5)M), and unaffected by CIP (10(-6)M). Collectively, these findings indicate that GIP exerts a moderate but statistically significant stimulatory effect on basal glucocorticoid secretion in rats, acting through specific receptors coupled with the adenylate cyclase/PKA dependent signaling pathway. PMID- 10465512 TI - Opposite effects on feeding of suprachiasmatic nucleus neuropeptide Y administration in rats. AB - The effects of injecting or infusing neuropeptide Y (NPY) into the suprachiasmatic nucleus of rats on patterns of individual macronutrient and water intake were examined during the following 2 h and also across 12 and 24 h light/dark cycles. Increased total energy intake (218 and 170%) and energy intake from the dextrin/sucrose diet (499 and 247%) were observed in the 2 h following injection of 100 pmol NPY at early light and early dark, respectively, and in the following 24 h (total energy: 67%, dextrin/sucrose: 73%). Nocturnal casein energy intake was also increased (258%) following NPY injection. Continuous infusion of 10 pmol/h of NPY suppressed nocturnal total energy (36%) and dextrin/sucrose intake (36%) as well as 24 h energy intake from casein (43%). These results demonstrate divergent effects of NPY subsequent to different mode of administration. PMID- 10465513 TI - Effects and interactions between alpha-MSH and MCH/NEI upon striatal cAMP levels. AB - It is known that alpha-MSH augments cAMP levels in rat brain slices containing accumbens and caudate-putamen nuclei. In this study we examined: a) the effect of other neuropeptides: MCH and NEI, on this cyclic nucleotide; b) if the effects of alpha-MSH on cAMP production can be modulated by addition of MCH or NEI to the incubation medium. Both MCH and NEI (3.6 microM) increased the production of cAMP, whereas at doses of 0.6 microM exerted no effects. When alpha-MSH 0.6 microM was added with NEI or MCH (0.6 microM), only MCH blocked the increase in the cAMP induced by alpha-MSH. Neither MCH nor NEI at the highest dose used (3.6 microM) had any additive effect on AMPc when added together with alpha-MSH. We conclude that, at a high concentration, (MCH/NEI)-like peptides can use the intracellular signal transduction linked to cyclic nucleotides in the CNS. PMID- 10465514 TI - Atrial natriuretic peptide effect on NADPH-diaphorase in rat intestinal tract. AB - Histochemical reaction of NADPH-diaphorase (NOS-NADPH-d) was used to identify NO synthesis. A 30-min 0.1 microg microg/kg/min ANP infusion led to about a 10% and 35% increase in small and large intestine enterocytes stain respectively. This increase was abolished by a bolus of 1 mg/kg L-NAME before ANP infusion in small intestine, and partially abolished it in colon. Incubation of small and large intestine with 0.5 microM ANP increased stain at about 20%. In both tissues the preincubation with 0.1 mM L-NAME abolished the ANP effect. Incubation with 0.1 mM 8-Br-cGMP enhanced staining about 70% and 30% in small and large intestine respectively. Our results show that ANP enhances NOS-NADPH-d activity, suggesting that ANP stimulates NO synthase in enterocytes by L-arginine-NO pathway. 8-Br cGMP mimicked the effect of ANP described above. Therefore, the guanylate cyclase coupled natriuretic receptors, NPR-A and NPR-B, probably mediate this ANP effect. PMID- 10465515 TI - Substance P evokes bradycardia by stimulation of postganglionic cholinergic neurons. AB - Substance P (SP) evokes bradycardia that is mediated by cholinergic neurons in experiments with isolated guinea pig hearts. This project investigates the negative chronotropic action of SP in vivo. Guinea pigs were anesthetized with urethane, vagotomized and artificially respired. Using this model, IV injection of SP (32 nmol/kg/50 microl saline) caused a brief decrease in heart rate (-30+/ 3 beats/min from a baseline of 256+/-4 beats/min, n = 27) and a long-lasting decrease in blood pressure (-28+/-2 mmHg from baseline of 51+/-5 mmHg, n = 27). The negative chronotropic response to SP was attenuated by muscarinic receptor blockade with atropine (-29 +/- 9 beats/min before vs -8 +/- 2 beats/min after treatment, P = 0.0204, n = 5) and augmented by inhibition of cholinesterases with physostigmine (-23 +/- 6 beats/min before versus -74 +/- 20 beats/min after treatment, P = 0.0250, n = 5). Ganglion blockade with chlorisondamine did not diminish the negative chronotropic response to SP. In another series of experiments, animals were anesthetized with sodium pentobarbital or urethane and studied with or without vagotomy. Neither anesthetic nor vagotomy had a significant effect on the negative chronotropic response to SP (F3,24 = 1.97, P = 0.2198). Comparison of responses to 640 nmol/kg nitroprusside and 32 nmol/kg SP demonstrated that the bradycardic effect of SP occurs independent of vasodilation. These results suggest that SP can evoke bradycardia in vivo through stimulation of postganglionic cholinergic neurons. PMID- 10465516 TI - SNV, a lipophilic superactive VIP analog, acts through cGMP to promote neuronal survival. AB - The current study explored whether the neuroprotective effects of vasoactive intestinal peptide (VIP) and its analog Stearyl-Nle17-VIP (SNV) were mediated through cGMP. SNV, was previously found to be 100-fold more potent than VIP in providing neuroprotection. Neuronal survival was assessed in rat cerebral cortical cultures. A cGMP antagonist (RP-8-pCPT-cGMPS, 10(-12)-10(-9) M) reduced the number of surviving neurons (40-60%), this decline was spared in the presence of SNV (10(-13)M). A cGMP agonist (Sp-8-pCPT-cGMPS, 10(-14)-10(-8)M) and SNV (10( 16)-10(-8)M) both provided significant neuroprotection against 10(-12) M of the cGMP antagonist. Immunoassays indicated that SNV induced increases in cGMP (two threefold) in these cultures, whereas VIP was 1000-fold less potent. These results implicate cGMP as a second messenger for VIP/SNV-mediated effects on neuronal survival. PMID- 10465517 TI - Immunohistochemical mapping of enkephalins, NPY, CGRP, and GRP in the cat amygdala. AB - This immunohistochemical study shows a wide distribution of neuropeptides in the cat amygdala. Neuropeptide Y is present along the whole amygdaloid complex, and fibers and cell bodies containing neuropeptide Y are observed in all the nuclei studied. Leucine-enkephalin-, gastrin-releasing peptide/bombesin-, and calcitonin gene-related peptide-immunoreactive fibers and perikarya are observed only in discrete nuclei of the amygdaloid complex, whereas only fibers -but no cell bodies- containing methionine-enkephalin-Arg6-Gly7-Leu8 have been observed. No immunoreactivity has been found for gamma-melanocyte-stimulating hormone, dynorphin A (1-17), or galanin. These data are compared with those reported in the amygdala of other mammals. PMID- 10465518 TI - Tuftsin: on the 30-year anniversary of Victor Najjar's discovery. AB - After a short description of the results of Victor Najjar's research on tuftsin and of the discoveries done by other authors in the early stage of tuftsin investigation, the current state of work on tuftsin is presented, based mainly on the literature published in the years 1984-1997. The presentation follows this order: the occurrence of tuftsin and retro-tuftsin sequences in proteins, their synthesis and biology, the antigenic properties of tuftsin, its influence on phagocytic cells, and other biologic activities of tuftsin, including antimicrobial, antiviral, antitumor and central effects, and the search for tuftsin superactive analogs. PMID- 10465519 TI - Mini symposium: the role of carbohydrates in reproduction. Introduction. PMID- 10465520 TI - Mammalian reproductive tract mucins. AB - Mucin glycoproteins are major constituents of the glycocalyx that covers mucosal epithelium. Two broad classes of mucins exist: membrane-associated and secretory. Of the secreted mucins, those with cysteine-rich regions are thought to polymerize through disulphide bonds. Among these gel-forming mucins are MUC2, MUC5AC, MUC5B and possibly MUC6. MUC7 lacks cysteine-rich domains and is thought to be secreted as a soluble monomer. Incomplete sequence information prevents classification of other mucins. Tandem repeats of amino acids rich in serine, threonine and proline are a common element in mucin core proteins, giving rise to relatively rigid, linear molecules with great potential for glycosylation. Ten distinct mucin genes have been identified in humans so far. Patterns of expression vary greatly. While MUC9, or oviductin, appears to be restricted to oviduct, the transmembrane mucin MUC1 is widely expressed. Proven functions for the different mucins are largely unknown, although potential functions are addressed in this review. Genetic and protein sequence information and expression profiles are also summarized, followed by a description of mucin assembly. Special attention is given to mucin expression in male and female reproductive tracts. PMID- 10465521 TI - Hyaluronan and proteoglycans in ovarian follicles. AB - Proteoglycans are macromolecules formed by a protein backbone to which one or more glycosaminoglycan side chains are co-valently attached. They can be secreted by the cells, retained at the cell surface, or stored in intracellular vacuoles. Hyaluronan is an extremely long glycosaminoglycan which, at variance with other glycosaminoglycans, is released into the extracellular matrix as a free polysaccharide not co-valently linked to a core protein. Both proteoglycans and hyaluronan influence many aspects of cell behaviour by multiple interactions with other molecules. They are involved in matrix formation, cell-cell and cell-matrix adhesion, cell proliferation and migration, and show co-receptor activity for growth factors. Both proteoglycan and hyaluranon synthesis change significantly during ovarian follicle development and atresia. This review describes the structure of these molecules and their possible function in ovarian physiology. PMID- 10465522 TI - The glycocalyx of the sperm surface. AB - The surface of mammalian spermatozoa is covered by a dense coating of carbohydrate-rich molecules forming a 20-60 nm thick glycocalyx. The majority of sugar residues are attached to proteins which are either integrated within the sperm membrane, or are more or less loosely associated with it. It is estimated that there may be several hundred different glycoproteins comprising the glycocalyx, some of which are synthesized within the testis. Others, however, are produced by the epithelia of the efferent ducts, epididymis and possibly other accessory glands, and become associated with the spermatozoa post-testicularly during transit through, and storage in, the male tract. The acquisition of the mature glycocalyx is associated with the attainment of full sperm fertilizing ability. Until its complete molecular structure is elucidated, the complex function of the glycocalyx remains obscure, though it may be related to membrane maturation and immunoprotection in the female tract, as well as to sperm-zona binding and fertilization. PMID- 10465523 TI - Carbohydrate-based interactions on the route of a spermatozoon to fertilization. AB - Male and female intercommunication along the route which the spermatozoon takes to fertilization utilizes the information potential of carbohydrates. A hierarchy of carbohydrate-based binding events exists ranging from spermatozoa-oviduct interaction to primary and secondary binding between spermatozoon and oocyte. Before in-vivo fertilization can occur, spermatozoa are stored in the caudal part of the isthmus, in tight contact with the epithelium cells lining the oviduct. The sperm reservoir seems to be created by surface-associated sperm lectins recognizing epithelial glycoconjugates. With the changing conditions in the oviduct at the time of ovulation, spermatozoa may shed those sperm lectins, creating new surfaces which allow spermatozoa to be released from the epithelium, complete capacitation and interact with the oocyte in the appropriate manner. The first contact between both gametes occurs at the spermatozoa-zona pellucida interface. The 'primary' binding initiates the acrosomal exocytosis of the spermatozoa, followed by the 'secondary' binding of the acrosome-reacted spermatozoon that in consequence leads to sperm penetration through the zona pellucida. Primary and secondary binding events are directed by the cooperative interactions of multiple carbohydrate-recognition systems that may act in a hierarchical and redundant manner. The current perspective will focus on the role of carbohydrate-binding sperm proteins in the sequence of binding events during fertilization in the pig. PMID- 10465524 TI - An elusive role for glycosylation in the structure and function of reproductive hormones. AB - The crescendo of events leading first to ovulation and subsequently to birth is orchestrated by a broad repertoire of hormones. The major hormones of the ovulatory cycle are representatives of four hormone classes: neurotransmitters, releasing factors, trophic hormones acting on target tissues, and steroid-like molecules released by the target tissues. The punctuate and staccato rhythm of the neurotransmitters and releasing hormones relentlessly drive the swelling and protracted wave of activity by the luteotrophic and steroid hormones. Carbohydrates alone are notably absent as hormones and the predominant role for glycosylation appears to be the conferment of increased solubility to endocrine molecules, either during their manufacture or by modulating circulatory half life. Rarely considered examples of the importance of glycosylation in reproductive hormones include adenosine, important for spermatozoan activity, and the hormone-binding globulins, which ensure the aqueous transport of hydrophobic steroids. The archetype glycoprotein hormones, especially human chorionic gonadotrophin (HCG), are discussed more extensively, as the structural and functional roles of carbohydrate in these hormones have been studied exhaustively. Conversely, the direct involvement of HCG and the importance of its carbohydrate for autonomous growth, in both placental invasion and tumorigenesis, has received little attention in the literature. PMID- 10465525 TI - Glycosylation of G-protein-coupled receptors for hormones central to normal reproductive functioning: its occurrence and role. AB - Many hormones that are central to normal reproductive functioning mediate their physiological effects by activating a receptor which belongs to the large family of G-protein-coupled receptors (GPCR). Members of this family of receptor proteins are usually glycosylated on extracellular domains. In recent years the role of this glycosylation in cell surface expression/protein folding, ligand recognition and receptor-effector coupling has been investigated. This review summarises current knowledge of the role of glycosylation in the functioning of the receptors for gonadotrophin-releasing hormone (GnRH), luteinizing hormone/human chorionic gonadotrophin (LH/HCG), follicle stimulating hormone (FSH), oxytocin (OT) and vasopressin (AVP). PMID- 10465526 TI - Transvaginal progesterone: evidence for a new functional 'portal system' flowing from the vagina to the uterus. AB - The results of many recent experimental and clinical studies support the hypothesis that progesterone administered vaginally is distributed selectively to the uterus where tissue concentrations and effects exceed expectations. This phenomenon has multiple clinical implications in several fields of gynaecological endocrinology, notably in assisted reproductive treatments and new forms of hormone replacement therapy. Yet, the actual mechanisms by which vaginal administration of progesterone can induce higher concentrations in the uterus, despite low concentrations in the systemic circulation, remain obscure and most puzzling to many gynaecologists. This review aims to muster ideas and propose different mechanisms to explain the observed phenomenon. In particular, we will summarize data that support the various putative modes of transport including, direct diffusion through tissue, intracervical aspiration, absorption into the venous or lymphatic circulatory systems and countercurrent vascular exchange with diffusion from utero-vaginal veins/lymph vessels to arteries. All these mechanisms may concur to various extents to the uterine specificity of vaginal progesterone. PMID- 10465527 TI - Hypothesis on the role of sub-clinical bacteria of the endometrium (bacteria endometrialis) in gynaecological and obstetric enigmas. AB - Unexplained infertility, recurrent abortion, dysfunctional uterine bleeding, pelvic pain, premenstrual syndrome, premature labour, placental insufficiency and pre-eclampsia are examples of common obstetric and gynaecological problems that frequently defy adequate explanation. Bacterial vaginosis, a non-inflammatory condition, is associated with premature labour, but antibiotics administered topically provide less effective prophylaxis than those administered orally. This would indicate that bacterial vaginosis might be a marker for significant genital tract bacteria, but some pathology is dependent on micro-organisms ascending out of reach of topical antibiotics. The author was led to consider the hypothesis that micro-organisms, possibly those associated with bacterial vaginosis, surreptitiously inhabit the uterine cavity (bacteria endometrialis) where they are culprits of some common gynaecological and obstetric enigmas. The objective of this review is to provide an initial theoretical examination of this hypothesis. Bacteria in the endometrium have been associated with infertility. Antiphospholipids have been linked to recurrent miscarriage and pre-eclampsia and with infections including Mycoplasma. Pre-eclampsia might be explained by an exaggerated host response to intrauterine micro-organisms or bacterial toxins. The hypothesis that one common factor, bacteria endometrialis, could provide a plausible explanation for a variety of obstetric and gynaecological mysteries is particularly intriguing. There is sufficient evidence to justify further investigation. PMID- 10465528 TI - Historical insights: John William Ballantyne 1861-1923. PMID- 10465529 TI - Novel conformationally restricted tetracyclic analogs of delta8 tetrahydrocannabinol. AB - Novel analogs of (-)-delta8-tetrahydrocannabinol (delta8-THC) in which the conformation of the side chain was restricted by incorporating the first one or two carbons into a six membered ring fused with the aromatic phenolic A ring were synthesized. The affinities of the novel ligands for CB1 and CB2 indicated that the "southbound" chain conformer retained the highest affinity for both receptors. PMID- 10465530 TI - On-resin macrocyclization of peptides via intramolecular SnAr reactions. AB - On-resin macrocyclization via an SNAr reaction is employed in the synthesis of tocinoic acid analogs. Specifically, an N-terminal nitrofluorobenzene is attacked by a nucleophilic C-terminal sidechain. The remaining nitro group can be reduced and acylated. NMR is used to compare the conformation of the new macrocyclic peptides to tocinoic acid. PMID- 10465531 TI - 9-Deoxopodophyllotoxin derivatives as anti-cancer agents. AB - Several 9-deoxo-9-substituted podophyllotoxin derivatives were synthesised starting from naturally occuring podophyllotoxin and their anti-cancer activity was evaluated against in vitro human cancer cell line assay. It was observed that these compounds do possess good anti-cancer activity particularly against ovarian, renal and lung cancer cell lines. PMID- 10465532 TI - Practical synthesis of all inositol stereoisomers from myo-inositol. AB - Synthesis of six inositol stereoisomers was successfully carried out via conduritol intermediates prepared from myo-inositol. Dihydroxylation and epoxidation followed by ring opening of the conduritol B, C and F derivatives gave epi-, allo-, muco-, neo-, DL-chiro- and scyllo-inositol. The cis-inositol derivative, which may not be prepared by this approach, was synthesized in 5 steps via 2-O-benzoyl-myo-inositol orthoformate as the key intermediate. PMID- 10465533 TI - Formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine under anaerobic conditions by reductively activated nitro 5-deazaflavin derivatives. AB - Electrolytically reduced 6- and 8-nitro-5-deazaflavin derivatives have been found to interact to react specifically with guanine base by means of cyclic voltammetry. Electrolytic reductions of 6- and 8-nitro-5-deazaflavin derivatives in the presence of the 2'-deoxyguanosine under anaerobic conditions resulted in prominent formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine. PMID- 10465534 TI - Highly sequence selective nonmacrocyclic two-armed receptors for peptides. AB - Simple nonmacrocyclic two armed receptors have been synthesized to create a new class of sequence-selective receptors for peptides. Screening several examples of these simple compounds against a 24,389-member library of N-acetyl tripeptides revealed novel binding properties. PMID- 10465535 TI - Structure-activity relationship of N-methyl-N-aralkyl-peptidylamines as novel N type calcium channel blockers. AB - Selective N-type voltage sensitive calcium channel (VSCC) blockers have shown efficacy in several animal models of stroke and pain. In the process of searching for small molecule N-type calcium channel blockers, we have identified a series of N-methyl-N-aralkyl-peptidylamines with potent functional activity at N-type VSCCs. The most active compound discovered in this series is PD 173212 (11, IC50 = 36 nM in the IMR-32 assays). SAR and pharmacological evaluation of this series are described. PMID- 10465536 TI - Casein kinase II inhibitors isolated from two Brazilian plants Hymenaea parvifolia and Wulffia baccata. AB - Two dihydroflavonol rhamnosides (1 and 2) isolated from the bark of Hymenaea parvifolia and two pentacyclic triterpenoids (3 and 6) obtained from the leaves of Wulffia baccata have been found to exhibit inhibitory effects of casein kinase II (CK-II) dose-dependently, suggesting that at higher doses more than 10 microM, these four compounds may act as potent CK-II suppressors of the CK-II-mediated activation of 60S acidic ribosomal P proteins in vitro. PMID- 10465537 TI - Disproportionation reaction of disulfides promoted by nitric oxide (NO) in the presence of oxygen. AB - Two disulfides brought about disproportionation reaction to afford an unsymmetrical disulfide in 50% yield with a catalytic amount of nitric oxide in the presence of oxygen. The reaction proceeded faster when alkyl disulfides were employed for the reaction, and the substituent effects suggested that the reaction commenced with an oxidative process. PMID- 10465538 TI - Derivatives of (R)-2-amino-5-methoxytetralin: antagonists and inverse agonists at the dopamine D2A receptor. AB - A series of N-arylmethyl substituted (R)-5-methoxy-2-(propylamino)tetralins has been prepared and evaluated for affinity and efficacy at dopamine (DA) D2A receptors. The novel compounds appeared to be antagonists or inverse agonists. (R)-2-[(Benzyl)propylamino]-5-methoxytetralin (7) was characterized as a potent inverse agonists at DA D2A receptors in a [35S]GTPgammaS binding assay. PMID- 10465539 TI - Synthesis of heterocyclic enol ethers and their use as group 2 metabotropic glutamate receptor antagonists. AB - Heterocyclic enol ethers of type 1 were studied with respect to the inhibition of 1S,3R-ACPD (10 microM)-stimulated GTP gamma35S binding on rat mGluR2 transfected cell membranes. The structure activity relationship with regard to the substitution pattern of the phenyl ring, the oxygen substituent and the nature of the heterocycle is discussed. PMID- 10465540 TI - Novel quinolizidine salicylamide influenza fusion inhibitors. AB - A novel series of quinolizidine salicylamides was synthesized as specific inhibitors of the H1 subtype of influenza A viruses. These inhibitors inhibit the pH-induced fusion process, thereby blocking viral entry into host cells. Compound 16 was the most active inhibitor in this series with an EC50 of 0.25 microg/mL in plaque reduction assay. The synthesis and the SAR of these compounds are discussed. PMID- 10465541 TI - Dimethyldioxirane epoxidation of polycyclic fluoranthene hydrocarbons. AB - Direct epoxidation of polycyclic fluoranthenes with dimethyldioxirane provides several new mono- and di-oxide derivatives. PMID- 10465542 TI - Design of peptides derived from anti-IgE antibody for allergic treatment. AB - We have designed and synthesized peptides derived from an anti-IgE antibody which has a potential for the treatment of allergy. It was indicated that conformational restriction of peptide via an intramolecular disulfide bond improved the binding affinity for IgE and that the peptide might have an ability to inhibit the IgE-receptor interaction. PMID- 10465543 TI - Structure-based design of irreversible, tripeptidyl human rhinovirus 3C protease inhibitors containing N-methyl amino acids. AB - Tripeptide-derived molecules incorporating N-methyl amino acid residues and C terminal Michael acceptor moieties were evaluated as irreversible inhibitors of the cysteine-containing human rhinovirus 3C protease (3CP). Such compounds displayed good 3CP inhibition activity (k(obs)/[I] up to 610,000 M(-1) s(-1)) and potent in vitro antiviral properties (EC50 approaching 0.03 microM) when tested against HRV serotype-14. PMID- 10465544 TI - Novel quinolinequinone antitumor agents: structure-metabolism studies with NAD(P)H:quinone oxidoreductase (NQO1). AB - The effects of functional group changes on the metabolism of novel quinolinequinones by recombinant human NAD(P)H:quinone oxidoreductase (NQO1) are described. Overall, the quinolinequinones were much better substrates for NQO1 than analogous indolequinones, with compounds containing heterocyclic substituents at C-2 being among the best substrates. PMID- 10465545 TI - Human chymase inhibitors based on the 1,2,5-thiadiazolidin-3-one 1,1 dioxide scaffold. AB - A series of compounds that utilize the 1,2,5-thiadiazolidin-3-one 1,1 dioxide scaffold was synthesized and shown to be highly effective inhibitors of recombinant human skin chymase. PMID- 10465546 TI - Synthesis of pyrazolidinone antibacterial agents. AB - The synthesis of monocyclic pyrazolidinones, which show moderate antibacterial activity, are described. PMID- 10465547 TI - SAR in the alkoxy lactone series: the discovery of DFP, a potent and orally active COX-2 inhibitor. AB - Extensive SAR has been established in the alkoxy lactone series and this has lead to the discovery of DFP (5,5-dimethyl-3-(2-propoxy)-4-methanesulfonylphenyl) 2(5H)-furanon e), a potent COX-2 inhibitor exhibiting in vivo efficacy in all models studied. PMID- 10465548 TI - Synthesis of a N-acylsulfamide linked dinucleoside and its incorporation into an oligonucleotide. AB - A N-acylsulfamide linked thymidine dinucleoside was synthesized and incorporated into an oligonucleotide (ON). The interest is in a linkage analog that has a higher pKa relative to a phosphodiester and when incorporated into ONs is capable of helix formation with complementary RNA. The hybridization property of the resultant ON with RNA was shown to result in significant destabilization. PMID- 10465549 TI - Nonpeptidic HIV protease inhibitors: 6-alkyl-5,6-dihydropyran-2-ones possessing a novel and achiral 3-(2-t-butyl-5-methyl-4-sulfamate)phenylthio moiety. AB - Dihydropyran-2-ones possessing a sulfamate moiety at the 4-position of the thiophenyl ring were designed to reach S3' pocket of the HIV protease. Synthetic routes for the preparation of thiotosylates possessing 3-(2-t-butyl-5-methyl-4 sulfamate) phenylthio moiety were established. SAR of various sulfamate analogs including HIV protease binding affinities, antiviral activities and therapeutic indices will be described. PMID- 10465550 TI - Triterpene and diterpene inhibitors of pyruvate dehydrogenase kinase (PDK). AB - Several oximes of triterpenes with a 17-beta hydroxyl and abietane derivatives are inhibitors of pyruvate dehydrogenase kinase (PDK) activity. The oxime 12 and dehydroabietyl amine 2 exhibit a blood glucose lowering effect in the diabetic ob/ob mouse after a single oral dose of 100 micromol/kg. However, the mechanism of the blood glucose lowering effect is likely unrelated to PDK inhibition. PMID- 10465551 TI - 7-Amino-4-methyl-6-sulfocoumarin-3-acetic acid: a novel blue fluorescent dye for protein labeling. AB - 7-Amino-4-methyl-6-sulfocoumarin-3-acetic acid (AMCA-S, also called Alexa 350) 2 was synthesized as a new water-soluble blue fluorescent dye for protein labeling. Compared with its nonsulfonated counterpart (AMCA) 1 the new dye gave significantly higher fluorescence quantum yields on proteins. PMID- 10465552 TI - 3-Alkyl-(5,5'-diphenyl)imidazolidineiones as new cannabinoid receptor ligands. AB - Twenty-four 3-alkyl-(5,5'-diphenyl)imidazolidinediones were synthesized and evaluated as new cannabinoid receptor ligands. Three compounds exhibited a Ki value around 100 nM against [3H]-SR 141716A binding obtained from human CB1 transfected CHO cells membranes. The lack of change of affinity in the presence of a non hydrolyzable GTP analogue seems to indicate they are cannabinoid antagonists. PMID- 10465553 TI - A 2-nitroimidazole carbamate prodrug of 5-amimo-1-(chloromethyl)-3-[(5,6,7 trimethoxyindol-2-yl)carbony l]-1,2-dihydro-3H--benz[E]indole (amino-seco-CBI TMI) for use with ADEPT and GDEPT. AB - The synthesis of a 2-nitroimidazol-5-ylmethyl carbamate prodrug 10 of the potent minor groove alkylating agent amino-seco-CBI-TMI 3 is described. Chemical, radiolytic, and enzymic reductions of a model 2-nitroimidazol-5-yl carbamate 8 show release of the amine effector upon reduction. Prodrug 10 gives a ten fold increase in cytotoxicity against human ovarian carcinoma SKOV3 cells in the presence of E. coli B nitroreductase (NTR) and a 21-fold increase in cytotoxicity against a SKOV3 cell line (SC3.2) transfected with the gene for NTR. The cytotoxicity of 10 increased 15- to 40-fold under hypoxia. Prodrug 10 has significant potential as a prodrug for use in ADEPT and GDEPT applications, and as a hypoxia-selective cytotoxin. PMID- 10465554 TI - Binding of Zn(II), Cu(II), and Fe(II) ions to Alzheimer's A beta peptide studied by fluorescence. AB - Binding of Zn(II), Cu(II) and Fe(II) ions to A beta1-40, A beta1-42 and a single tryptophan mutant of Abeta 1-40 in solution at pH 7.4 was studied by fluorescent titration. Job plots and fitting of titration curves revealed formation of 1:1 and 1:2 peptide-metal complexes. For dimeric peptides A beta1-40 and A betaF4W the order of metal to peptide affinities is Fe < Cu > Zn, which is in agreement with the Irving-Williams series of complex stability. The affinity of A beta1-42 for Fe increases dramatically upon aggregation: K(D) changes from ca. 100 to ca. 0.2 microM. PMID- 10465555 TI - Photonuclease activity of Taylor's blue. AB - Taylor's blue (1,9-dimethylmethylene blue, DMMB+) associates with DNA, at least in part, through intercalation as is evidenced from the red shift in the absorption maximum, diminution of the fluorescence, and induced circular dichroism in the presence of nucleic acid. Irradiation of DMMB+/covalently closed circular supercoiled phiX174 phage DNA complex at lambda > 520 nm leads to DNA nicking in a dose-dependent manner. PMID- 10465556 TI - Targeting nitric oxide to cancer cells: cytotoxicity studies of glyco-S nitrosothiols. AB - Glyco-S-nitrosothiols, fructose-2-SNAP and glucose-2-SNAP, were synthesized and found to be much more cytotoxic than SNAP in killing DU-145 human prostate cancer cells in vitro. PMID- 10465557 TI - Increased antiviral activity of cyclic urea HIV protease inhibitors by modifying the P1/P1' substituents. AB - A series of alkyl substituted P1/P1' analogs was prepared in an attempt to increase translation of the 3-aminoindazole class of HIV protease inhibitors. Increasing the lipophilicity of the P1/P1' residues dramatically improved translation of enzyme activity to antiviral activity in the whole cell assay. PMID- 10465558 TI - Ring E analogs of methyllycaconitine (MLA) as novel nicotinic antagonists. AB - We have prepared ring E analogs of the diterpenoid alkaloid methyllycaconitine. These compounds have been assayed for nicotinic activity and were found to act as functional antagonists on adrenal nicotinic receptors. PMID- 10465559 TI - Significant compensatory role of position Y-2 conferring high affinity to non phosphorylated inhibitors of Grb2-SH2 domain. AB - Systematic modification of amino acid at position Y-2 of a library-derived non phosporylated thioether-cyclized peptide, cyclo(CH2CO-Glu2-Leu-Tyr0-Glu-Asn-Val Gly-Met-Tyr-Cys) -amide, aided by molecular modeling, demonstrates that the Glu( 2) sidechain compensates for the absence of Tyr0 phosphorylation in retaining effective binding to Grb2-SH2 domain. Replacement of Glu(-2) with gamma carboxyglutamic acid produced a high affinity inhibitor, the first example with submicromolar affinity (IC50 = 640 nM). PMID- 10465560 TI - Synthesis and biological evaluation of a new class of vaccine adjuvants: aminoalkyl glucosaminide 4-phosphates (AGPs). AB - A novel series of acylated omega-aminoalkyl 2-amino-2-deoxy-4-phosphono-beta-D glucopyranosides (aminoalkyl glucosaminide 4-phosphates) was synthesized and screened for immunostimulant activity. Several of these compounds enhance the production of tetanus toxoid-specific antibodies in mice and augment vaccine induced cytotoxic T cells against EG.7-ova target cells. PMID- 10465561 TI - Novel protein kinase C inhibitors: synthesis and PKC inhibition of beta substituted polythiophene derivatives. AB - A series of beta-substituted polythiophene derivatives was synthesized through palladium-catalyzed coupling reaction. Their structure-protein kinase C (PKC) inhibitory activity relationship was studied. The carboxaldehyde and hydroxymethyl derivatives of alpha-terthiophene were potent PKC inhibitors (IC50 = 10(-7) M). PMID- 10465562 TI - Total synthesis of the potent proteasome inhibitor epoxomicin: a useful tool for understanding proteasome biology. AB - Epoxomicin (1), a peptide alpha',beta'-epoxyketone isolated from the actinomycete strain No.Q996-17, possesses potent in vivo anti-tumor and anti-inflammatory activities. In this paper, we report the first syntheses of epoxomicin, [3H] epoxomicin, and a biotinylated epoxomicin analog as well as the absolute configuration of the epoxide stereocenter. The natural product and derivatives have permitted the first identification of the proteasome as the specific cellular target of epoxomicin. PMID- 10465563 TI - Effects of ischemia on QT dispersion during spontaneous anginal episodes. AB - Myocardial ischemia induced by pacing, angioplasty, or stress results in a significant increase in QT dispersion (QTd = QT maximum - QT minimum). This study investigated the effects of ischemia on QTd and the rate-corrected QTd (QT(c)d) during spontaneous anginal episodes in patients with coronary artery disease (CAD). Ninety-five patients with CAD and typical angina pectoris and 15 control subjects complaining of anginalike symptoms were studied. QTd and QT(c)d were calculated from 12-lead surface electrocardiograms recorded during and after the relief of pain. QTd and QT(c)d were significantly higher during the anginal episode (84+/-31 ms and 98+/-51 ms) compared to the painless conditions (69+/-24 ms and 71+/-24 ms) (P = .003 and P = .001 for QTd and QT(c)d, respectively) only in the 57 CAD patients who had a history of an old previous myocardial infarction. QTd and QT(c)d are significantly increased during spontaneous angina in patients with documented CAD and history of previous myocardial infarction. PMID- 10465564 TI - Spatial distribution of QT dispersion measured by magnetocardiography under stress in coronary artery disease. AB - This study investigated changes in spatial distribution of QT duration in patients with and without coronary artery disease (CAD) using magnetocardiography. Thirty-six-channel magnetocardiograms (MCGs) were registered at rest and under stress in 15 patients with chest pain, seven of whom had significant coronary stenosis. QT dispersion (QTd) was calculated for MCG and 12 lead electrocardiogram (ECG) under both conditions. For MCG, homogeneity of repolarization was measured using a smoothness index (SI). Also, at each registration site, the intraindividual difference between QT at rest and under stress was determined (deltaQT). QTd values as determined by standard 12-lead configurations were not significantly different between groups. MCG QTd values were significantly higher in the CAD group at rest only when all available channels were taken into consideration (P < .05); SI values differed significantly between groups under both conditions (rest, P < .005; stress, P < .01). Good separation between groups was possible using the range of deltaQT (P < .05) and SI (deltaQT) (P < .005). Consideration of the spatial distribution of QTd increases its sensitivity in the detection of CAD, suggesting that CAD involves complex changes in repolarization, not apparent in limited lead sets such as standard 12-lead configurations. PMID- 10465565 TI - Dispersion of the monophasic action potential duration in patients with polymorphic ventricular tachycardia. AB - The mechanism of polymorphic ventricular tachycardia (PMVT) remains unclear. To investigate the electrophysiologic mechanism of PMVT, monophasic action potentials (MAPs) were recorded with a contact electrode technique from right ventricular sites during sinus rhythm and right ventricular pacing. MAPs were obtained from 6 patients with PMVT (PMVT group) and 11 patients without PMVT (control group). The duration from the onset of the upstroke to 90% repolarization of the MAP (MAPD90) during right ventricular pacing at both pacing cycle lengths of 600 and 400 ms was significantly longer in the PMVT group than in the control group (332+/-60 ms vs 279+/-33 ms [P < .005] and 276+/-32 ms vs 229+/-23 ms [P < .0001], respectively). Dispersion of the MAPD90 in sinus rhythm was significantly larger in the PMVT group than in the control group (52.5+/-34.6 ms vs 26.1+/-12.0 ms [P < .005]), and dispersion of the MAPD90 during right ventricular pacing at both pacing cycle lengths of 600 and 400 ms was also significantly larger in the PMVT group than in the control group (86.0+/-44.2 ms vs 37.4+/-28.6 ms [P < .005], and 48.8+/-19.3 ms vs 27.1+/-7.1 ms [P < .05], respectively). Dispersion of repolarization time (activation time plus MAPD90) at a pacing cycle length of 600 ms was longer in the PMVT group than in the control group (104.3+/-38.9 ms vs 49.4+/-31.2 ms [P < .05]). These results suggest that the patients with PMVT have a greater regional dispersion of ventricular repolarization time and that the heterogeneity of ventricular repolarization may play an important role in the genesis of PMVT. PMID- 10465566 TI - An interactive graphical system for automated mapping and display of cardiac rhythms. AB - Electrical cardiac mapping has been used to study the mechanisms of cardiac arrhythmias, to assist in clinical diagnosis of rhythm disorders, to guide interventional procedures, or to evaluate the effects of antiarrhythmic drugs. Manual determination of local activation times, the first step in constructing activation maps, is a time-consuming process that precludes the possibility of on line interactive mapping during an electrophysiologic experiment or a clinical procedure. This report describes an interactive graphic user interface application that (1) automatically determines the activation sequences with good accuracy, (2) displays graphical presentations of activation maps within seconds, and (3) allows manual interactive adjustments. Five automated activation time detection algorithms, one for bipolar electrograms and four for unipolar electrograms, were evaluated and compared. High-density canine recordings were used to evaluate the accuracy of the system. Data included normal atrial activation (sinus rhythm) and abnormal reentrant atrial activation (atrial flutter). PMID- 10465567 TI - Assessment of right artrial mapping and P wave-triggered signal-average in patients with paroxysmal atrial fibrillation. AB - To assess right atrial mapping and P wave-triggered signal-averaged electrocardiogram (ECG) in patients with paroxysmal atrial fibrillation (PAF), this study examined right atrial electrograms using atrial mapping and parameters by P wave-triggered signal-averaged ECG in 39 patients without sick sinus syndrome. Subjects were divided into those with PAF (n = 13; 60+/-13 years old) and a control group (n = 26; 49+/-19 years old). The total number of abnormal right atrial electrograms per patient was significantly greater in the PAF group (3.2+/-1.9) than in the control group (1.1+/-0.9; P < .001). The longest duration of right atrial electrogram in the PAF group tended to be greater than that in the control group (P = .06). The filtered P wave duration was significantly longer in the PAF group than in the control group (144+/-21 vs 125+/-14 ms [P < .002]). The values of the root mean square of P wave-triggered signal-averaged ECG 15 ms from the onset (RMSi 15) and 20 ms from the offset (RMSe 20) were significantly lower in the PAF group (1.1+/-0.4 microV, 1.4+/-0.5 microV) than in the control group (1.9+/-1.1 microV [P < .02], 2.1+/-0.9 microV [P < .01]). The total number of right atrial electrograms in patients with RMSi 15 of < or =1.5 microV was significantly greater than in patients with RMSi 15 of >1.5 microV (2.2+/-1.8 vs 1.3+/-1.3 [P < .05]). Thus, the total number of abnormal right atrial electrograms per patient, the total filtered P wave duration, RMSi 15, and RMSe 20 may be good indices of PAF in patients without sick sinus syndrome. RMSi 15 may reflect the total number of the abnormal right atrial electrograms per patient. PMID- 10465568 TI - Body surface potential field representation fidelity: analysis of map estimation procedures. AB - The first part of this study analyzed the spatial-temporal error distribution of the Lux-type limited lead system. Quantitative new evidence is reported that the 32-lead anterior subset estimates the further 160 leads with an average amplitude error less than 38.5 microV. The spatial error distribution revealed 8 sites where the error is the highest, primarily on the anterior side, independent of the clinical classification. The second part of the study examined inter-lead system conversion strategies for interpolating the Lux-192 lead maps from the Montreal-63 data. The methodology based on the Laplacian interpolation yielded an average amplitude error of 143.7 microV and an average correlation of 0.87 for pattern fidelity. In this specific case a modified linear interpolation surpassed the Laplacian method. A presented example illustrates that even in cases when the fidelity of the signal information is heavily compromised the diagnostic information may remain less influenced. PMID- 10465569 TI - Heart rate variability and left ventricular dilatation early after myocardial infarction. AB - To assess clinically whether alterations of autonomic tone precede left ventricular dilatation, heart rate variability and early left ventricular dilatation after a first myocardial infarction were assessed. Low-frequency power (LF), high-frequency power (HF), and total power (TP) were obtained by ambulatory electrocardiogram on day 1 in 53 patients with a first acute myocardial infarction. Left ventricular end-diastolic volume determined by echocardiography was obtained on day 1 and day 14. Stepwise linear regression analysis was used to assess the associations of early left ventricular dilatation with heart rate variability adjusted for clinical variables. Higher LF and TP were significantly associated with early left ventricular dilatation after adjustment for age, sex, site of myocardial infarction, acute revasucularization, peak creatine kinase level, history of hypertension, and use of angiotensin-converting enzyme inhibitors and beta-blockers. Higher LF and TP preceded early left ventricular dilatation after myocardial infarction. PMID- 10465570 TI - Electrocardiographic patterns of patients with echocardiographically determined biventricular hypertrophy. AB - The numerous criteria proposed for the electrocardiographic (ECG) diagnosis of biventricular hypertrophy (BVH) suffer from inadequate correlative data. We used two-dimensional (2D) echocardiography to identify BVH and analyzed the ECG patterns in these patients. The study group had 69 such patients with BVH and the control group had 22 patients with isolated left ventricular hypertrophy (LVH) demonstrated by 2D echocardiography. The electrocardiograms were analyzed for the presence of established criteria used in the diagnosis of LVH and right ventricular hypertrophy (RVH). Of the 69 patients in the study group, 17 (25%) had ECG findings of BVH, 25 (36%) had LVH, and 14 (20%) had RVH. An S wave in V5/V6 of >7 mm was most the frequent finding in the 17 patients with BVH on the electrocardiogram. The sensitivity of ECG criteria for BVH was 24.6%, specificity was 86.4%, and positive predictive value was 85%. This study reemphasizes the difficulty of ECG diagnosis of BVH. The electrocardiogram has a low sensitivity but satisfactory specificity and positive predictive accuracy for BVH. PMID- 10465571 TI - Prevalence and outcome of ST-segment elevation in posterior electrocardiographic leads during acute myocardial infarction. AB - The prevalence and clinical significance of ST-segment elevation (STE) in posterior electrocardiographic (ECG) leads during acute myocardial infarction (AMI) is largely unknown. We obtained posterior ECG leads (V7, V8, and V9), as well as standard 12-lead and right precordial leads (V4R-V6R), immediately upon admission in 210 consecutive patients with AMI. Nineteen patients (9% of 210 cases) had STE of > or =1 mm in 2 or more posterior leads, either as an isolated finding (7 cases, 3.3% of 210) or in association with STE at inferior or lateral sites (12 cases, 5.7% of 210). STE in posterior leads was detected in 10.9% (7 of 64) of patients without STE in standard ECG leads. Tall R waves in V1-V2 developed in 5 cases (26.3% of 19). Patients with STE in posterior leads were significantly older (64.4+/-7.3 years vs 58.9+/-8.9 years) with more frequent cardiovascular risk factors (3.4+/-1.1 vs 2.9+/-1.2) than those without. In hospital complications were significantly more frequent in these patients compared with matched controls (Mantel-Haenszel odds ratio = 7, confidence interval = 1.28-28.43). There was also a trend toward lower ejection fraction and increased in-hospital mortality that did not reach statistical significance. We conclude that STE in posterior ECG leads is not uncommon among patients with AMI and no STE in standard leads and may portend a worse in-hospital course. PMID- 10465572 TI - Right bundle branch block as a cause of false-negative ECG classification of inferior myocardial infarction. AB - It is generally accepted in clinical electrocardiography that a right bundle branch block (RBBB) does not interfere with the electrocardiographic (ECG) diagnosis of myocardial infarction (MI). The basic assumption is that the initial excitation wavefronts are relatively unchanged in RBBB. This study compared serial changes in Q wave duration in inferior leads II, III, and aVF in 9 patients who developed RBBB within 3 weeks after myocardial revascularization procedure (RBBB group) and in 41 revascularized patients without RBBB in the same observation period (control group). Q wave durations in the electrocardiograms obtained before the patients' procedures were not significantly different between the study and control groups. However, Q wave durations shortened significantly more in the RBBB group than in the control group. The most pronounced Q wave duration shortening took place in lead aVF, -18.2 ms in the RBBB group versus 3.8 ms in the control group (P = .0001). The shortening was less pronounced, although significant, in leads II and III: II, -7.6 +/- -10.9 ms in the RBBB group vs -2.3 +/- -3.5 ms in the control group (P = .01); III, -11.3 +/- -10.5 ms vs -2.6 +/- -6.5 ms (P = .002); aVF, -18.2 +/- -13.5 ms vs -3.8 +/- -5.3 ms (P < .0001). It is concluded that incident RBBB complicating revascularization procedures may cause significant alterations in spatial orientation of the initial excitation wavefronts. This may be a potential source of false-negative ECG diagnosis of inferior MI, particularly in clinical trials where serial ECG analysis is an important part in MI classification. PMID- 10465573 TI - Impending acute myocardial infarction during severe exercise associated with a myocardial bridge. AB - A young man had an impending acute myocardial infarction while playing soccer. Chest pain and anterior ST-segment elevation lasted 3 hours despite anti-ischemic medication, including streptokinase thrombolysis. An electrocardiogram recorded after the symptoms had passed was normal. There was a minimal increase in cardiac enzyme levels, and a pyrophosphate scan and echocardiogram were normal. Coronary cineangiography showed normal coronary arteries except for systolic compression of the left anterior descending coronary artery. An exercise stress test, while the patient was on atenolol, showed absence of myocardial ischemia. This impending acute myocardial infarction could have been caused by an acute thrombus with lysis prior to catheterization or by a deep muscle bridge that kinked or twisted the coronary artery due to myocardial forceful muscular contraction during the sympathetic stimulation of exercise. In conclusion, an impending acute myocardial infarction may occur in young patients having myocardial bridges, and a beta-blocker must be administered, especially when this condition appears during severe exercise. PMID- 10465574 TI - Prominent bifid T waves observed in the QT prolongation caused by complete atrioventricular blockade in a hypokalemic diabetic patient. AB - A 63-year-old diabetic man was admitted with general fatigue. Electrocardiogram (ECG) on admission showed complete atrioventricular (AV) blockade associated with prominent bifid T waves. The second component of the bifid T waves was distinguished from U waves by the beat-to-beat varying bifidity and the nadir between the two components located at > or = 1 mm above the isoelectric line. Range of absolute QT interval was 535 to 650 ms. Hypokalemia (3.6 mEq/L) was noted at admission. Partial restoration of the potassium level (3.9 mEq/L) prior to temporary ventricular demand pacing obscured the bifid T waves and attenuated the QT prolongation and dispersion to some extent (absolute QT interval ranging 520 to 620 ms). It was concluded that marked bradycardia caused by complete AV blockade (ie, a junctional escaped rhythm at a rate of 42 beats/min), hypokalemia, and underlying diabetes mellitus contributed in concert to the QT prolongation and dispersion leading to the prominent bifid T waves. PMID- 10465575 TI - Consensus on the management of malignant melanoma of the skin in The Netherlands. Dutch Melanoma Working Party. AB - In 1996 the Dutch Melanoma Working Party, in co-operation with the National Organization for Quality Assurance in Hospitals in the Netherlands and the Dutch Association of Comprehensive Cancer Centres, organized the third consensus conference on the management of melanoma of the skin. The following guidelines were approved. The recommended margin for diagnostic excision is 2 mm of macroscopically normal skin around the lesion; the margins for therapeutic excision are 1 cm of normal skin for a Breslow thickness of < or = 2 mm and 2 cm for a Breslow thickness of > 2 and < or = 4 mm. A margin of at least 2 cm also appears to be justified for thicker melanomas. Elective lymph node dissection is not recommended. Sentinel node biopsy appears to be a promising method to detect occult metastases in the regional lymph nodes. If regional lymph node metastases are present, therapeutic regional lymph node dissection must be conducted. Isolated regional perfusion is indicated for inoperable tumour growth in an extremity. Radiotherapy can be applied curatively (for example, if surgery is not possible), palliatively (if desired in combination with hyperthermia) or postoperatively (if non-radical resection is suspected). Adjuvant systemic therapy for melanoma patients is still experimental. Atypical (dysplastic) naevi and congenital naevi are major risk factors for melanoma. No consensus has been reached about the prophylactic excision of all congenital naevi. A follow-up period of 5 years is sufficient for patients with a melanoma of < or = 1.5 mm Breslow thickness (provided there are no histological signs of regression) and of 10 years when the Breslow thickness is > 1.5 mm. The patient should be actively involved in the follow-up (inspection, palpation). Regular routine blood tests, radiological examination and ultrasound scanning are not considered to be worthwhile. There is no evidence that the growth of micro-metastases is stimulated by hormonal changes during pregnancy or contraceptive pill use. Excessive exposure to ultraviolet radiation should be discouraged. Regular population screening for melanoma is not considered to be worthwhile, owing to the relatively low frequency and the predominantly favourable stage at the time of diagnosis, particularly in young people. PMID- 10465576 TI - Transcription of the MAGE-1 gene and the methylation status of its Ets binding promoter elements: a quantitative analysis in melanoma cell lines using a real time polymerase chain reaction technique. AB - The human MAGE gene family comprises at least 12 highly homologous genes. This makes it very difficult to assess expression of a single member quantitatively by means of Northern blotting. In order to investigate expression of the MAGE-1 gene quantitatively we therefore used the recently developed real-time polymerase chain reaction (PCR), a novel fluorescence-based quantitative PCR technique. This powerful technique enables detection of expression levels which differ by as much as a factor of 10(5) in magnitude. MAGE-1 expression is known to correlate with demethylated status of the Ets binding sites of its promoter. In a panel of 19 melanoma and nine non-melanoma cell lines we were able to confirm the relationship between MAGE-1 expression and demethylation of the Ets binding promoter region. Five cell lines, however, showed only very slight expression, while the two essential Ets promoter elements were largely demethylated. Earlier studies have shown that treatment of MAGE-1-negative cell lines with the demethylating agent 5-aza-2'-deoxycytidine (DAC) is sufficient to induce MAGE-1 expression. We were able to induce clear MAGE-1 expression in two of the non expressing cell lines by incubation with DAC, although this expression did not reach very high levels. Consistent with this low level of induction is the observation that the Ets binding sites of the MAGE-1 promoter were not completely demethylated in the DAC-treated cell populations. In conclusion, we show in this study that the real-time PCR technique is a very useful tool for the quantification of expression of highly homologous genes. PMID- 10465577 TI - CD44 variant isoform CD44v10 expression of human melanoma cell lines is upregulated by hyaluronate and correlates with migration. AB - CD44, a multifunctional adhesion receptor involved in cell-cell and cell-matrix interactions, plays an important role in the local progression and metastasis of malignant tumours. We investigated relative CD44 variant isoform mRNA expression in six human melanoma cell lines and determined cell migration on hyaluronic acid (HA) coated substrates. Haematopoietic form (CD44H) mRNA expression increased in all melanoma cell lines after plating on HA, whereas the relative CD44 variant exon 10 (CD44v10) mRNA expression increased in only three of the cell lines. Cell migration rates increased on substrates coated with HA in the three CD44v10 positive cell lines, whereas the three CD44v10-negative cell lines showed no modification in migration rates. Immunofluorescent labelling of CD44v10 revealed increased expression with plaques localized to the periphery of cells. Cell lines with increased relative CD44v10 expression exhibited significantly higher mean migration rates on HA. These results indicate that CD44v10 expression functionally relates to melanoma cell migration and suggest that interaction between CD44v10 and HA plays a role in the variable tissue invasion and aggressiveness of different melanoma clones. PMID- 10465578 TI - In vitro and in vivo comparison between the effects of treatment with adenosine triphosphate and treatment with buthionine sulfoximine on chemosensitization and tumour growth of B16 melanoma. AB - In this study we compare the effects of treatment with external sodium adenosine 5'-triphosphate (ATP) with the effects of L-buthionine-SR-sulfoximine (BSO) on B16 melanoma growth and on the modulation of the cytotoxic antimelanoma activity of cyclophosphamide (CY). We evaluated the in vitro effects of treatment with ATP or BSO on intracellular glutathione (GSH) levels, mitochondrial membrane potential (delta psi(m)) and the proliferation rate of the B16F10 melanoma cell line. Compared with untreated cells, delta psi(m) and GSH levels were already significantly decreased (25% and 57% reduction, respectively) after the first hour of incubation in culture cells exposed to 3 mM ATP. After 24 and 48 h a major reduction was observed in delta psi(m) (nearly 30%). GSH levels were also maximally depleted at 24 h (approximately 75%) and partially recovered (up to 37% of levels of control) after ATP was removed from the medium. At 24 and 48 h, the proliferation rate was decreased 1.4- and 1.7-fold, respectively, compared with control cells. Treatment with 50 microM BSO produced a time-dependent decrease in GSH levels (0.5, 21, 48 and 97.3% reduction at 1, 4, 8 and 24 h, respectively), but up to 54% of the levels of control cells was recovered after BSO was removed from the medium. In contrast to ATP, neither delta psi(m) nor proliferation rate was significantly modified in the first 24 h with BSO treatment. At 48 h, delta psi(m) was reduced by nearly 27%, and cell proliferation decreased 1.2-fold compared with controls. When the in vitro cytotoxic effect of low dose acrolein (an active metabolite of CY) in combination with BSO or ATP was analysed, a synergistic effect was found between BSO and acrolein, with a dose modification factor (DMF) of 1.98, but the antiproliferative effects of acrolein plus ATP were only approximately additive (DMF = 1.05). In addition, in in vivo studies differential effects were found between ATP and BSO. Specifically, whereas BSO alone significantly increased the survival time of mice bearing B16 melanoma liver metastases, and enhanced the cytotoxic effect of CY on this tumour model, no therapeutic benefits could be observed with ATP treatment, either alone or in combination with diethyl maleate (a GSH-depleting agent) and CY. In conclusion, our findings show that in our experimental system, both extracellular ATP and BSO have growth-inhibitory properties against B16 melanoma in vitro. In vivo, however, only BSO produces a chemosensitizing effect, whereas ATP has not proved useful as a biological modifier of chemotherapy. PMID- 10465579 TI - Cytostatic activity of coumarin metabolites and derivatives in the B16-F10 murine melanoma cell line. AB - Coumarin has antitumour effects in vivo and cytostatic effects in vitro. Its half life in humans is short (1-1.5 h) and the monohydroxylated biotransformation products have significantly longer half-lives. One or several of these products may thus be responsible for the antitumoral effects. We have assayed the in vitro cytostatic activity of five monohydroxylated coumarins (3-, 4-, 6-, 7- and 8 monohydroxycoumarin), their acetates and methyl-ethers. Murine melanoma cells (cell line B16-F10) and murine fibroblasts (B82) were exposed to the test compounds at concentrations between 10 and 160 microg/ml. The cytostatic effects were estimated by reduction of the tetrazolium dye MTT. In the melanoma cells, some of the compounds inhibited growth after exposure for 1 day. In contrast, coumarin inhibited growth to a smaller extent, and only after exposure for 3 days. The most active compounds (3-acetoxycoumarin, 4-methoxycoumarin and 6 hydroxycoumarin), as well as coumarin, were also assayed in murine fibroblasts. The cytostatic effects of 4-methoxycoumarin and 6-hydroxycoumarin were less pronounced in fibroblasts than in melanoma cells. Our observations suggest that these compounds may have a greater therapeutic margin. PMID- 10465580 TI - Mutation analysis of the MEN1 tumour suppressor gene in malignant melanoma. AB - During the initiation and progression of malignant melanoma a series of genetic events accumulate, including alterations of chromosome 11q. Recently, an important tumour suppressor gene, the multiple endocrine neoplasia type 1 (MEN1) gene, has been mapped on 11q13 and has been cloned. To assess whether the MEN1 region is involved in tumour initiation and progression, we analysed 23 primary cutaneous melanomas and 17 metastases for loss of heterozygosity (LOH) using two informative polymorphic markers closely linked to the MEN1 gene (PYGM and D11S449). To search for mutations within the gene, single-strand conformation polymorphism (SSCP) analysis was performed using 13 primer sets with designed intronic sequences to amplify the MEN1 coding sequence exons 2 to 10. None of the cases showed LOH at the MEN1 gene locus. By SSCP analysis, no aberrant bands were identified on exons 3 to 10. Analysis of exon 2 revealed the presence of aberrant bands in two of the analysed melanomas. Sequencing analysis revealed a genetic polymorphism at S145S (AGC-->ACT) in both sections. None of the cases analysed showed MEN1 gene mutations. This study represents the first genetic analysis of the MEN1 gene in sporadic melanomas. Our data demonstrate no evidence of deletion or mutation of the MEN1 gene in primary or metastatic melanoma. Therefore, MEN1 gene alterations appear not to be associated with tumorigenesis of malignant melanoma. The MEN1 gene appears to be a highly specific tumour suppressor gene only involving tumours within the spectrum of MEN1 disease. PMID- 10465581 TI - Intercellular adhesion molecule-1 (ICAM-1) and granulocyte-macrophage colony stimulating factor (GM-CSF) co-expression in cutaneous malignant melanoma lesions. AB - The expression of intercellular adhesion molecule-1 (ICAM-1) and granulocyte macrophage colony stimulating factor (GM-CSF) was investigated in 25 melanoma patients by evaluating 34 fresh biopsy specimens. ICAM-1 in situ hybridization and immunochemistry for ICAM-1 and GM-CSF were performed. Most of the metastatic melanoma samples (12 out of 18) and a few of the primary melanoma lesions (three out of 16) showed ICAM-1 expression. The expression of ICAM-1 was significantly (P < 0.01) higher in metastatic lesions than in primary tumours. GM-CSF mRNA and protein were detected in 10 of the 18 metastatic samples and in two of the 15 primary lesions. A significantly high degree (P < 0.0002) of concordance between ICAM-1 and GM-CSF expression was observed: the samples that were negative or positive for ICAM-1 expression were correspondingly negative or positive for GM CSF. Correlation with clinical and histological parameters was examined. The expression of both molecules in metastatic samples was found to be significantly (P < 0.001) associated with a shorter recurrence-free period. These findings, if confirmed by a wider number of patients, could suggest the prognostic value of the simultaneous, and probably co-ordinated, expression of ICAM-1 and GM-CSF. They also highlight the importance of preventive molecular and biochemical characterization of neoplastic cell cytokine receptors, specifically focusing on the particular cytokine to be used as anticancer therapy and/or as adjunct to chemotherapy. PMID- 10465582 TI - Differential increase of Fas ligand expression on metastatic and thin or thick primary melanoma cells compared with interleukin-10. AB - Fas ligand (FasL), a cell surface molecule belonging to the tumour necrosis factor family, binds to its receptor Fas and thus induces apoptosis of Fas bearing cells such as activated lymphocytes. In this paper, we report the expression of FasL on melanoma cell lines and patient tumour specimens, and compare it with the expression of interleukin-10 (IL-10), a putative immunosuppressive factor. Apoptosis of Fas-bearing Jurkat cells was increased after interferon-alpha treatment of the FasL-positive melanoma cell line A375, suggesting a regulation of FasL function. We also tested whether FasL and IL-10 were ever co-expressed. Immunohistochemistry studies showed that IL-10 expression was highly positive in the same tumour samples which expressed FasL. In the melanoma patients with thin primaries, 10 of the 12 primaries and six of the seven metastatic lesions were positive for IL-10. In the melanoma patients with thick primaries (> 0.75 mm), four of the five primary lesions and nine of the 10 metastatic lesions were positive for IL-10. In contrast, FasL was generally negative in primary tumours and positive in metastatic tumours. In the thin primary melanoma patients, two of the 12 primaries and five of the seven metastatic tumours were positive for FasL. From the thick melanomas, one of the five primaries and five of the 10 metastatic lesions were positive for FasL. The function of melanoma-derived FasL was confirmed by four different cytotoxicity assays. PMID- 10465583 TI - Prognostic value of tumour vascularity in primary melanoma. AB - To investigate the prognostic value of tumour vascularity we studied 84 patients with primary melanomas ranging in tumour thickness (Breslow) from 0.37 to 7 mm and in depth of tumour infiltration (Clark) from II to V. Vascularization was assessed by immunohistochemistry with a CD-31 antibody recognizing endothelial cells. The CD-31-positive vessels were counted and the degree of vascularization was correlated with the survival of the patients. In addition, the relationship between blood vessel density and some histopathological data is discussed. In our study, the multivariate Cox model showed that the only independent variable in disease-free survival was tumour thickness (Breslow classification) and the only one in overall survival was depth of tumour infiltration (Clark classification). In disease-free survival, tumour thickness (Breslow classification) was a clear prognostic factor (P = 0.004) after 4 years' follow-up, as were depth of tumour infiltration (Clark classification) (P = 0.04) and ulceration (P = 0.04). In overall survival, tumour vascularity was the strongest prognostic factor at 4 years, high vascularity being associated with a good prognosis (P = 0.06). Clark classification was also a prognostic factor (P = 0.02) in overall survival. We conclude that high vascularization is associated with a better prognosis but is not an independent prognostic indicator. PMID- 10465584 TI - Overexpression of mutant ras in human melanoma increases invasiveness, proliferation and anchorage-independent growth in vitro and induces tumour formation and cachexia in vivo. AB - Malignant melanoma is the deadliest form of skin cancer. Previous studies have shown that the incidence of ras mutation increases with progression of melanoma, but that such mutations may not be present in the earliest radial growth phase melanomas. Recently it has been proposed that introduction of ras mutations into cells deficient in tumour suppressor genes such as p16 (INK4a) is sufficient to induce characteristics of cellular transformation such as anchorage-independent growth and tumour formation in vivo. To test this hypothesis in human melanoma, mutant N-ras, mutant H-ras or wild-type H-ras genes were transfected by electroporation into WM35 cells, a p16-deficient human melanoma cell line of low invasive potential. Increased expression of mutant ras p21 enhanced anchorage dependent cell growth on tissue culture plastic. In addition, overexpression of mutant N-ras and H-ras, but not of wild-type H-ras, increased the experimental invasive potential, inducing anchorage-independent growth in soft agar, increasing cell motility measured by time-lapse video microscopy, and increasing invasiveness through reconstituted basement membranes. Finally, overexpression of mutant H-ras in melanoma cells was shown to increase tumorigenicity and to induce cachexia when H-ras transfected cell lines were injected subcutaneously in severe combined immunodeficiency (SCID) mice. Thus the addition of activating ras mutations to a melanoma cell line already deficient in p16 leads to enhanced proliferation, survival and migration in vitro and to enhanced subcutaneous tumour formation in vivo. This phenotype is typical of the behaviour of vertical growth phase (VGP) melanoma, and we propose that activation of the ras signalling pathway in the presence of deletions in p16 or related tumour suppressors can induce the VGP melanoma phenotype. PMID- 10465585 TI - Mutagen sensitivity in common cutaneous malignant melanoma and dysplastic naevus syndrome. AB - Sunlight (ultraviolet radiation) has been identified as the major environmental risk factor for the development of cutaneous malignant melanoma and dysplastic naevi. This is, however, not sufficient to explain all melanoma cases. In recent years much emphasis has been given to genetic susceptibility to melanoma. A biomarker of susceptibility to environmentally related cancer is mutagen sensitivity. This is measured as the number of chromatid breaks in lymphocytes which are exposed to bleomycin in the G2 phase of the cell cycle. It has been described that patients with common melanoma show an increased mutagen sensitivity compared with controls. In the present study mutagen sensitivity was measured in 10 dysplastic naevus syndrome patients and compared with that in 11 patients with common melanoma. We found similar results for common melanoma patients as have been reported earlier: a relatively high mean breaks per cell value (0.93 +/- 0.31). In contrast, melanoma patients with dysplastic naevi showed a significantly (P<0.01) lower mutagen sensitivity value (0.46 +/- 0.34). This phenomenon was even more pronounced when only hereditary dysplastic naevi patients (one or more family members with dysplastic naevi) were considered (n = 5; 0.24 +/- 0.05). These results suggest a difference in the initiation of the carcinogenic process in melanoma with a dysplastic naevus as a precursor and melanoma without dysplastic naevi. PMID- 10465586 TI - Effects of transpupillary thermotherapy on immunological parameters and apoptosis in a case of primary uveal melanoma. AB - Transpupillary thermotherapy (TTT) is a new treatment modality for uveal melanoma. We studied whether application of TTT influences the immunogenicity of the tumour cells in vivo or the expression of molecules related to apoptosis. Immunohistochemistry using monoclonal antibodies directed against HLA molecules, HMB45, P53, Fas ligand (FasL), Fas, Bcl-2 and tumour-infiltrating cells was applied to sections of an enucleated eye containing a uveal melanoma that received TTT 1 week before enucleation. The innermost part of the tumour which had been exposed directly to the laser treatment showed no staining for HLA antigens, nor for Fas or FasL epitopes. The intermediate part of the tumour showed a wet necrosis and HLA expression similar to the expression in the peripheral tumour. A large number of macrophages were observed in the necrotic as well as the intact tumour tissue, especially bordering the wet necrotic area. FasL and Bcl-2 were only expressed in the viable, outer part of the tumour. This immunological evaluation of one case of uveal melanoma treated with TTT revealed that TTT may not only have a direct destructive effect on the primary tumour, but may also influence the immunogenicity of uveal melanoma cells, induce infiltration of macrophages into the tumour, and induce apoptosis. The presence of many macrophages suggests that they play a role in the removal of the TTT treated tumour tissue by phagocytosis. PMID- 10465587 TI - Nodular histogenetic type--the most significant factor for thick melanoma: implications for prevention. PMID- 10465588 TI - Conference report 3rd International Conference Adjuvant Therapy of Malignant Melanoma. PMID- 10465589 TI - Ensuring compliance with NRMP policy. PMID- 10465590 TI - Functions of newly identified members of the tumor necrosis factor receptor/ligand superfamilies in lymphocytes. AB - Rapid progress in the discovery of members of the tumor necrosis factor receptor/ligand superfamilies has been made, mainly with the help of massive DNA sequencing and bioinformatic studies. Biological studies of the new members have not only indicated overlapping roles with other members but also provided insights into novel functions. In particular, multiple pairings of receptors and ligands highlight a complex control mechanism of immune responses by these superfamilies. PMID- 10465591 TI - [Primary dysfunction of the graft in liver transplantation: is it possible to predict this complication?]. PMID- 10465592 TI - Routine screening of children returning home from the tropics. Authors do not prove case for routine screening. PMID- 10465593 TI - Comparison of inhaled beclometasone and budesonide. Back titration of inhaled steroids is uncommon in New Zealand. PMID- 10465594 TI - Comparison of inhaled beclomethasone and budesonide. Study was inadequate. PMID- 10465595 TI - Comparison of inhaled beclomethasone and budesonide. Studies of potencies of asthma drugs have methodological limitations. PMID- 10465596 TI - Why heart disease mortality is low in France. Authors' hypothesis is wrong. PMID- 10465597 TI - Pressures of acute obstetrics on consultants. Reply from Royal College of Obstetricians and Gynaecologists. PMID- 10465598 TI - Usefulness of contacting other experts when conducting literature searches. Study only "proves" what librarians knew anyway. PMID- 10465599 TI - Cyber triage. Calls forwarded. PMID- 10465600 TI - Report cards. Cleveland's Phoenix. PMID- 10465601 TI - Needle safety. Unstuck in California. PMID- 10465602 TI - Food service. Culinary rehab. PMID- 10465603 TI - Job shifts. Good counsel. PMID- 10465605 TI - Should CEO grow, then go? PMID- 10465604 TI - Do HMOs purposely stall in paying claims? PMID- 10465606 TI - Innovators. Kinder cuts. PMID- 10465607 TI - Mentors. Binding them with science. PMID- 10465608 TI - Healtheon + WebMD. Hype helps, but success sells. PMID- 10465609 TI - Personality tests. PMID- 10465610 TI - Theory and simulation. Can theory challenge experiment? PMID- 10465611 TI - Antihistamines: back to the future. Preface. PMID- 10465612 TI - Antihistamines: back to the future. Summary of the conclusions. BSACI. British Society for Allergy and Clinical Immunology. PMID- 10465613 TI - Management of scorpion sting. PMID- 10465614 TI - Is BNP ready for use in clinical practice? PMID- 10465615 TI - Transfusion associated graft versus host disease. PMID- 10465616 TI - Outpatient clinics for adults with congenital heart disease. PMID- 10465617 TI - International Congress on Vascular Medicine. Graz, Austria, 25-27 June 1998. Abstracts. PMID- 10465618 TI - Proceedings of the 5th International Symposium on Clinical Evaluation of Drug Efficacy in Urinary Tract Infection and of the Urinary Tract Infections Symposia of the Commission for Treatment of Urinary Tract Infection of the International Society of Chemotherapy. 29 June-3 July 1997. Sydney, Australia. PMID- 10465619 TI - Guess what! Keratosis lichenoides chronica. PMID- 10465620 TI - Guess what! Tropical phagedenic ulcer. PMID- 10465622 TI - Literature review. PMID- 10465621 TI - Guess what! Eruption of lymphocyte recovery. PMID- 10465623 TI - Image of the month. Bouveret's syndrome. PMID- 10465624 TI - Image of the month. Peutz-Jeghers syndrome. PMID- 10465625 TI - Image of the month. Sister Mary Joseph's nodule: carcinoma of the pancreas with umbilical metastasis. PMID- 10465626 TI - Image of the month. Fascioliasis. PMID- 10465627 TI - Image of the month. Lymphoma involving the pancreaticoduodenal and para-aortic lymph nodes. PMID- 10465628 TI - Image of the month. Hereditary angioedema (HAE). PMID- 10465629 TI - When does heartburn warrant more concern? PMID- 10465630 TI - E-biomed: improving access or a threat to peer review? PMID- 10465631 TI - Attractin more interest in weight regulation. PMID- 10465632 TI - A 37-year-old patient with human immunodeficiency virus (HIV) disease. PMID- 10465633 TI - Expression of leptin and its receptor in the human stomach. PMID- 10465634 TI - National resident matching program. PMID- 10465635 TI - Humoral immunity in acute and chronic hepatitis C infection. PMID- 10465636 TI - ECL cell tumor and endocrine carcinoma of the stomach. PMID- 10465637 TI - Duplex ultrasonography and TIPS. PMID- 10465638 TI - Rebound hypersecretion after omeprazole. PMID- 10465639 TI - A case of infective endotipsitis. PMID- 10465640 TI - Regression of MALT lymphoma of the rectum after anti-H. pylori therapy in a patient negative for H. pylori. PMID- 10465641 TI - Balloon endoscopic sclerotherapy, a novel treatment for high-risk gastric fundal varices: a pilot study. PMID- 10465642 TI - Nonsurgical treatment modalities for chronic anal fissure using botulinum toxin. PMID- 10465643 TI - The results of a major demonstration and evaluation program undertaken by HCFA in 1989: the Medicare Alzheimer's Disease Demonstration and Evaluation Program (MADDE). PMID- 10465645 TI - Characterization of each isoform of a F(ab')2 by capillary electrophoresis. AB - Free solution capillary electrophoresis was investigated for the characterization of an M(r) 100 000 purified F(ab')2. Optimization of the experimental conditions allowed the identification of five separated peaks, suggesting the presence of isoforms which differed by only 0.2 pH unit. This heterogeneity was still detectable with 80 amol of protein. After a preparative separation by chromatofocusing, identification of each form was performed for the first time by capillary electrophoresis. A quantitative and qualitative correlation with isoelectric focusing showed that free solution capillary electrophoresis represents a sensitive method for revealing subtle differences in charge, even for large proteins. PMID- 10465647 TI - Determination of gestational age from lunar age assessments in human fetal remains. PMID- 10465646 TI - Evaluation of aspartic acid racemization ratios in the human femur for age estimation. PMID- 10465644 TI - Chimeric forms of furin and TGN38 are transported with the plasma membrane in the trans-Golgi network via distinct endosomal pathways. AB - Furin and TGN38 are menbrane proteins that cycle between the plasma membrane and the trans-Golgi network (TGN), each maintaining a predominant distribution in the TGN. We have used chimeric proteins with an extracellular Tac domain and the cytoplasmic domain of TGN38 or furin to study the trafficking of these proteins in endosomes. Previously, we demonstrated that the postendocytic trafficking of Tac-TGN38 to the TGN is via the endocytic recycling pathway (Ghosh, R.N.,W.G. Mallet,T.T. Soe,T.E.McGraw, and F.R. Maxfield.1998.J. Cell Biol.142:923-936). Here we show that internalized Tac-furin is delivered to the TGN through late endosomes, bypassing the endocytic recycling compartment. The transport of Tac furin from late endosomes to the TGN appears to proceed via an efficient, single pass mechanism. Delivery of Tac-furin but not Tac-TGN38 to the TGN is blocked by nocodazole, and the two pathways are also differentially affected by wortmannin. These studies demonstrate the existence of two independentpathways for endosomal transport of proteins to the TGN from the plasma membrane. PMID- 10465648 TI - [Surgical procedures that can be conducted on an ambulatory basis. Intranasal release of paranasal sinus cysts]. PMID- 10465649 TI - EBNA-1 sequences in endemic and sporadic Burkitt's lymphoma. PMID- 10465650 TI - An ethnopharmacological survey for potential angiotensin converting enzyme inhibitors from Indian medicinal plants. AB - Plants used in different traditional systems of Indian medicine and some relatives/substitutes have been investigated for their angiotensin converting enzyme (ACE) inhibitory activity. They were selected on the basis of their usage as cardiotonics, diuretics and other uses related to the symptoms of hypertension. Out of the 73 species investigated, 22 showed more than 50% ACE inhibitory activity. PMID- 10465651 TI - Medicinal and veterinary plants of El Caurel (Galicia, northwest Spain). AB - A study of the medicinal and veterinary plants popularly used in El Caurel region (Lugo province, northwest Spain) and their relationships is reported. We obtained data for 85 species belonging to 31 families of vascular plants. Their vernacular names, properties, preparations, mode and popular uses are presented. PMID- 10465652 TI - Studies on the immunomodulatory effects of Boerhaavia diffusa alkaloidal fraction. AB - The alkaloidal fraction of Boerhaavia diffusa was studied for its effect on cellular and humoral functions in mice. Oral administration of the fraction (25 100 mg/kg) significantly inhibited SRBC-induced delayed hypersensitivity reactions in mice. However, the inhibition was observed only during post immunisation drug treatment, while no effect during pre-immunisation drug treatment was observed. A significant dose-related increase in antibody titre was observed during pre- and post-immunisation treatment. The alkaloidal fraction failed to show any blastogenic responsiveness of murine splenocytes to Concanvalin A (Con A) and lipopolysaccharide (LPS). Similarly, it did not display any mitogenic activity. Thus, the present study has shown the in vivo immunostimulatory activity of B. diffusa alkaloidal fraction without an in vitro effect. PMID- 10465653 TI - Effects of aqueous extracts of Hibiscus macranthus and Basella alba in mature rat testis function. AB - Mature male albino Wistar rats (180-220 g) were given by gastric intubation Hibiscus macranthus Hochst A ex Rich (Malvaceae) and Basella alba L. (Basellaceae) aqueous extract from both fresh and dry leaves, at a dose equivalent to 0.720 or 0.108 g of plant, respectively per kg body weight. This was to evaluate their effects on male reproductive function. Control groups were treated equally, but given water instead of the extract. After the treatment periods, animals were killed, their blood collected, the testes and some annex glands removed for histological and biochemical analysis. Results showed that the extract from fresh leaves significantly increased the body weight of rats by 17% from day 7 as compared to controls, whereas the increase was less pronounced (4%) when the rats were given dry leaf extract. The weight of seminal vesicles of rats given the extracts also increased after 15 days. The histological analysis of testis showed abundant spermatozoa in the lumen of the seminiferous tubulus from day 7 in rats fed with the extract when compared to the controls. The serum level of testosterone was significantly increased on the 15th day by 80% in rats given both types of extracts compared to the controls. Testis of treated rats showed high testosterone production in vitro (136 and 62%, respectively for treated and control after 15 days, compared to those of 3 days). Activity of prostatic acid phosphatase was high in prostate, testis and serum of treated rats in all experimental period. From these findings and observation, it was concluded that the aqueous extract of H. macranthus and B. alba had anabolizing and virilizing effects. PMID- 10465654 TI - Antiinflammatory effect of the aqueous extract from Rumex patientia L. roots. AB - Pharmacological studies were conducted with the aqueous extract of roots of Rumex patientia L. (Polygonaceae) on experimental animals. For evaluating the antiinflammatory activity, carrageenan, histamine, dextrane, serotonine formaldehyde-induced oedema tests, cotton-pellet granuloma, and Kabak tests in rats were used. The extract was found to possess antiinflammatory activity. Acute toxicity studies revealed that the extract up to a dose of 3 mg/kg orally was nontoxic. PMID- 10465655 TI - Chemical and pharmacological evaluation of different ayurvedic preparations of iron. AB - Ayurvedic preparations of metallic iron commonly categorised as different 'putas' of 'Louha Bhasma' was chemically analysed and pharmacologically investigated in iron deficiency anemia. Atomic absorption spectral (AAS) study of different putas of Louha Bhasma revealed the presence of various proportions of important metals along with varied concentration of iron in it. The effect of a representative puta viz. 50 puta of Louha Bhasma in the management of agar gel diet and phlebotomy induced iron deficiency anemia in animal model was found to be statistically highly significant (P < 0.001) in comparison to the control and standard drug Fefol treated groups. PMID- 10465656 TI - The effects of Securidaca longepedunculata root extract on ionic currents and contraction of cultured rat skeletal muscle cells. AB - The effects of the primary extract roots of Securidaca longepedunculata were tested on sodium, calcium and potassium currents in rat skeletal muscle cells developed in culture. In addition, they were tested on depolarisation-induced contraction and resting intracellular calcium levels. S. longepedunculata extract (10(-6) g/l) increases sodium current at all potentials. No clear effect was observed on calcium current except for a slight increase at negative potentials ( 30, -10 mV) revealing a 5 mV shift towards negative potentials of the I(Ca)/V curve, as with potassium current. In contrast, at the same concentration, S. longepedunculata enhanced the contractile response elicited by durable depolarisation. This was not attributable to the slight increase in resting intracellular free calcium concentration which did not change during and following S. longepedunculata application. These results strongly suggest that S. longepedunculata root extract contains one or more components acting on the voltage-sensor of excitation-contraction coupling (dihydropyridine receptors), regardless of its implication as a calcium channel. PMID- 10465657 TI - The medicinal flora of Similipahar forests, Orissa State, India. PMID- 10465658 TI - Effect of isoorientin isolated from Arum palaestinum on uterine smooth muscle of rats and guinea pigs. AB - The phytochemical investigation of Arum palaestinum resulted in the isolation of two flavone C-glucosides, namely isoorientin (luteolin 6-C-glucoside) and vitexin (apigenin 8-C glucoside). The effects of isoorientin on rat isolated aorta, ileum, trachea and uterus and on guinea-pig uterus were studied. Isoorientin (10( 7)M-6 x 10(-4)M) caused concentration-dependent inhibition of the amplitude and the frequency of the phasic contractions of the rat and guinea-pig uterus but did not affect the isolated aorta, ileum or trachea. The results were discussed in relation to the effects of its aglycone luteolin reported in the literature. PMID- 10465659 TI - Chemistry and pharmacology of Monimiaceae: a special focus on Siparuna and Mollinedia. AB - The chemistry and pharmacology of species of the family Monimiaceae are reviewed, with special attention given to the genera Mollinedia and Siparuna, the two most important and representative in Brazil. The isolation of benzylisoquinoline alkaloids and kaempferol derivatives from Siparuna apiosyce is reported, as well as the isolation of aporphines from the fruits of Siparuna arianeae. Cinnamic acid derivatives and a gamma-lactone were isolated from Mollinedia gilgiana and Mollinedia marliae. PMID- 10465660 TI - Origins of APHA's Medical Care Section. American Public Health Association. PMID- 10465661 TI - Chaodynamic loss of complexity and self-similarity in cancer. AB - 'Omnis cellula a cellula': each cell descends from another cell--and most complex organ structures are built up of multiple cells at least. Even the underlying mechanisms of physiological as well as pathological processes reflect complexity in a pronounced manner. Loss of complexity, however, has been detected in aging and apparently also in the case of a number of diseases. This paper describes the loss of complexity in both carcinogenesis and the related growth pattern of cancer. After disruption of the cell's stability as a result of mutations, a decrease in the ability of the cell to induce a self-organized response is associated with the loss of control parameters at the cellular and molecular level. Complexity, understood as a principle of order, is altered in certain malignant tumors, as is indicated by a loss of the golden mean and by the disappearance of self-similarity. The lack of self-similarity could, therefore, be an ideal practical marker of malignancy in medical image analysis. PMID- 10465662 TI - On the vital role of fluid movement in organisms and cells: a brief historical account from Harvey to Coulson, extending the hypothesis of circulation. AB - The importance of the orchestrated movement of substrates over multicatalytic enzymes, topographically arranged for maximum efficacy for metabolic function, means that the organization of the body right down to the subcellular level is based upon the principle of perfusion for most normal living processes. The history of the development of this idea, which is contrary to the received wisdom arising and still persisting since the early days of biochemistry, will be followed from the time when the overall need for a circulation at the gross level was fully appreciated, extending the argument through a historical perspective to include the control of flow within the individual cell. Life, seen from this new perspective, seems to be inherently dependent upon this principle to such an extent that it leads to a clearer distinction between the living and the dead, and provides a framework for more critical experiments concerning control mechanisms in the cell. PMID- 10465663 TI - Another look at: fuel + O2 --> CO2 + H2O. Developing a water-oriented perspective. AB - Water plays many vital roles which must be maintained despite constant threat of water stress from the environment. Mechanisms to maintain these roles over the long-term involve intermediates of many metabolic pathways, such as glycerol, sucrose, free amino acids and their derivatives. In addition to viewing metabolic products as intermediates along energy-related pathways, nutrition researchers and epidemiologists should consider them as determinants of intracellular fluid maintenance. Satiety processes, oxidative-fuel selection, hormonal control, and intracellular-signaling processes can all be interpreted in terms of water oriented metabolism. Water intake and hydration status can vary considerably, and may be inadequate for a sizeable proportion of Western adults. Due to the metabolic adaptations required to compensate for perpetually inadequate water, chronic sub-optimal water intakes may be highly relevant to chronic disease etiology. Preliminary evidence links water-oriented metabolism to obesity, diabetes, cardiovascular disease, hypertension and cancer. Development of a water oriented perspective may reveal an important new area of research in human nutrition and epidemiology. PMID- 10465664 TI - Genetic induction and upregulation of cyclooxygenase (COX) and aromatase (CYP19): an extension of the dietary fat hypothesis of breast cancer. AB - A novel model of mammary carcinogenesis is proposed involving sequential induction and upregulation of cyclooxygenase and aromatase genes by essential fatty acids prominent in the US diet. The basic carcinogenic processes are: (1) constitutive prostaglandin biosynthesis and formation of mutagenic oxygen and nitrogen free radicals responsible for tumor initiation; (2) PGE-2-induced expression of aromatase and constitutive estrogen biosynthesis which sustains mitogenesis and tumor promotion; and (3) PGE-2-induced expression of vascular endothelial growth factor which stimulates angiogenesis and tumor metastasis. PMID- 10465665 TI - Microbes and sequestered substances as mechanisms for disease: Bartonella and L forms as common global etiological agents. AB - The pathogenicity of microbes may be determined by substances sequestered from blood and bound to their constituent lipid. The brain may not perceive substances sequestered by microbes, to interfere with control to maintain normal levels. Pathological conditions can be induced as organisms exposed to antimicrobial substances/conditions and/or deprived of nutrients essential to cell wall synthesis, disintegrate to free lipid-bound compounds and produce L-forms that can deplete nutrients as they revert to bacteria. Microbes may act as active carriers for the continuing interaction of sequestered substances. Changes in the molecular structure of substances effected during sequesteration could cause them to be seen as substances 'synthesized' by an organism. In media that contain substances to inhibit 'contaminants', L-forms can be seen as mycoplasma. Elementary bodies of L-forms with a specific substance or tissue affinity may be seen as 'receptors'. Bartonella are global agents for disease--pleomorphic organisms (description suits Proteus)--and they can be seen as 'contaminants'. PMID- 10465666 TI - Glycoaminoglycan (GAG) deficiency in protective barrier as an underlying, primary cause of ulcerative colitis, Crohn's disease interstitial cystitis and possibly Reiter's syndrome. AB - Ulcerative colitis, Crohn's disease and interstitial cystitis share many common features, the most important of which is a defect in the glycoaminoglycan (GAG) defensive barrier. This defect allows penetration of toxins causing localized inflammatory response, followed by fibrosis and distant pathological changes, together with a myriad of biochemical and immunological changes. The latter has caused confusion as to etiology of the aforementioned disorders. This hypothesis is somewhat supported by the fact that agents such as glucosamine and pentosan polysulphate (Elmiron) that replace the GAG layer, improve the conditions. The potential for extrapolation of this hypothesis to atherosclerosis and arthropathies exists. There is a great danger in modern medical research that if one misses the wood for the trees, one becomes hopelessly lost in the minutiae of research. At present, it is embarrassing that ulcerative colitis (UC), Crohn's (CR) and interstitial cystitis (IC) are the cause of a great deal of morbidity and occasionally mortality, yet after intensive research, the etiology and effective treatment eludes us. The research in the past has focused extensively on inflammatory response in the mucosal lining, and biochemical, infective and immunological changes in the serum. This has led to a vast array of research pathways that seem at the present time to be totally lost and, might I say, aimless in direction, as a cause for these conditions, that remain amongst the most imperically treated in modern medicine. Another possible syndrome in this class would be Reiter's, which has many features in common with the above. The basic tenet of a GAG deficiency hypothesis is that, as shown in Figure 1A, an intact GAG layer provides, firstly, a mechanical and electrostatic defence against penetration of infective agents, toxins, antigenic protein moieties, etc. and, secondly, the prevention of extravasation of body fluid components. A degraded GAG layer is the start of the disease cascade of the above group of illnesses. PMID- 10465667 TI - Can mirrors alleviate visual hemineglect? AB - Following right hemisphere stroke, many patients display an indifference to objects and events in the left side of the world ('neglect'). Here, we describe a new technique that might help accelerate recovery from neglect. The patient sits at a table and a mirror is propped vertically on the patient's right side in the parasagittal plane, so that when the patient rotates his head rightward and looks into the mirror, he sees the neglected side of the world reflected in the mirror. Our question was: since the sensory information was now coming from the non neglected left side, would this somehow make him overcome the neglect? In pilot experiments, two types of responses were seen: (a) In one subset of patients the presence of the mirror seemed to enhance the patients' awareness of the neglected field, so that they reached correctly for an object that was shown in the neglected field. Will repeated practice with this task accelerate recovery from neglect? (b) The second group of patients kept reaching into the mirror to grasp the reflection or kept groping behind the mirror ('mirror agnosia'). If the mirror was placed in the coronal position and the object placed behind their head, then some of these patients (from group B) reached correctly for the object. Quite apart from its obvious theoretical implications, we believe this technique might provide a new approach for the treatment of visual hemineglect. PMID- 10465668 TI - Evidence for an enhanced flow of virtual energy in the progression from inanimate matter and its role in behaviour proper to the animate state. AB - There are reasons for cleaving a virtual or phase space component from electromagnetic radiation and for reposing in this component properties of specific value for the functioning biosystem. Putative mechanisms are derived by which a stream or streams of virtual energy dissipates biomatter, providing global control of component chemical reactions. Characteristics include a variety of wavelengths whose interactions can change rapidly producing a collapse restoration cycle of values, an exquisite sensitivity to external fields of magnetic, gravity and heat origin plus a property for electron transport from lesser to more dense streams. Disturbances in the virtual control stream over chemical events become available as resonances to macroscopic measuring tools including those of clinical use. PMID- 10465669 TI - Sudden infant death syndrome and early family interpersonal relationships. AB - The medical profession are concerned with diseases or disorders, their diagnoses, and the appropriate treatment of their patients. The relevant processes involve interpersonal relationships in physical examinations, associated with the conventional social ritual courtesies. There are only two certainties--birth and death--with areas of probabilities and uncertainties in between. One such uncertainty is the sudden death of an infant in its first year. After the obstetrical birth processes, the specific health of the neonate involves the paediatrician. His skill and modern technical resources have enabled numbers of the prematurely born to survive. Has the modern neonate evolved over the centuries to have some needs that must be worked out between mother and her baby? To attribute hands-on experiences to genes takes away four holding hands from what they feel, the eyeball movements and what they see, the noses and what they smell, and the breast milk for the energy to move the muscles no longer in floating weightlessness. PMID- 10465670 TI - Tumor necrosis factor-alpha: a continuum of liability between insulin-dependent diabetes mellitus, non-insulin-dependent diabetes mellitus and carcinoma (review). AB - In this review, tumor necrosis factor-alpha (TNF-alpha) is identified as the uniting principle linking the pathogenesis of insulin-dependent diabetes mellitus (IDDM), non-insulin dependent diabetes mellitus (NIDDM) and carcinoma. Elevated TNF-alpha initially increases, and then inhibits, the activity of a number of key enzymes involved in energy metabolism and major histocompatibility (MHC) class I molecule expression. These enzymes include: protein-tyrosine kinase (PTKase) and protein-tyrosine phosphatase (PTPase--enzymes involved in energy metabolism, cell proliferation and stimulation of the MHC class I molecule pathway. Of primary importance is the inhibiting effect of TNF-alpha on PTKase, since this induces insulin resistance in NIDDM and carcinoma, and PTPase, which inhibits MHC class I molecule expression. Studies have shown that IDDM is associated with an increase in PTPase activity which leads to overexpression of MHC class I molecules and a concomitant destruction of pancreatic beta cells. Conversely, carcinoma is associated with an inhibition of PTPase activity, which reduces the expression of MHC class I antigen expression on the cell surface thereby allowing malignant cells to escape immune surveillance. It will be argued that there is continuum of liability between these three conditions, initiated by the effect of TNF-alpha on these key enzymes. PMID- 10465671 TI - No man is an island; no island is an island: does the immune network extend beyond the limits of skin? AB - Now that we are moving towards a post-antibiotic era, it may be necessary to reevaluate our concept of immunology. A long-standing handicap may be that, thus far, we have focused exclusively on the individual and his own immune machinery. A search for community interactions, also involving microflora, may provide clues for our survival under the extremely unhygienic conditions of the past as well as the present. Among the many adverse factors, could there also be hidden beneficial effects as well? If so, the development of such factors to therapeutic significance would be a worthwhile challenge. Diverse historical chronicles and a certain resistance shown by infants in shanty towns, after the maternal immune protection has faded and until their own immune system is fully developed, led us to postulate that adaptation is a prerequisite for any subject in order to benefit from unhygienic crowded conditions. PMID- 10465672 TI - Endothelin-secreting tumors and the idea of the pseudoectopic hormone secretion in tumors. AB - Ectopic hormone secretion in tumor cells is here described as an amplification of hormone production already present in normal, nonendocrine tumor-originated tissue. This idea is tested on the available data regarding endothelin-1 (ET-1) secreting tumors. The endothelins are ubiquitous regulatory peptides produced by various tissues. The precursor cells of many tumor types secrete endothelins. ET 1 protein expression was detected in situ in all tested prostate cancers as well as in normal prostate tissue. The majority of hepatocellular carcinomas produce ET-1, while ET-1 is secreted by the normal hepatic stellate cells. Human breast cancer cells produce immunoreactive ET-1. Similar data exist for pancreatic tissue, the thyroid and large bowel. We can conclude that tumor cells might sustain endothelin secretions already present in the normal tumor-originated tissue. The model that is presented of the pseudoectopic hormone secretion consists of relations between a few parameters. The proportion of hormone secreting tumors (Th) among all tumors (T) of that organ depends on the amount of the hormone-secreting cells (Ch) among all cells (C) susceptible to malignant transformation. The corrective factor (k) was introduced in the expression Th/T=Ch/C*k, to represent specific conditions altering the malignant transformation probability for a certain normal hormone-secreting cell. In prostate, breast and colon, the kvalue is predicted to be approximately 1, suggesting that ET-1-secreting normal cells are not more prone to the malignant transformation than their neighbours. In liver and pancreas, the incidence of ET 1-secreting tumors outnumbers the proportions of normal ET-1-secreting cells (k values >1). In these organs, normal ET-1-secreting cells seem more likely to turn malignant in comparison to their neighbours, perhaps due to their function, position and exposition to oncogenic factors, or even due to their ET-1 secretion. There are similar data for thyroid and adrenal glands. No ET-1 secretion was reported in kidney neoplasms. Normal renal ET-1 secreting cells might be less prone to turn malignant than other renal cells. Unlse the normal lung tissue, small cell lung cancers often secrete adrenocorticotrophic hormone (ACTH). The pancreatic islet cells do not secrete gastrin, but their tumors often do. Constant k would exceed 1 in both cases. We speculate that these tumors might originate from a small subset of cells with the described feature. Tumor cells sometimes lack features of the normal tissue, as in the cases of the steroid receptor-negative breast cancer. These tumors might originate from the hypothetical subset of receptor-free breast cells. Benign breast epithelial cells lacking oestrogen receptors have been described in cases of megalomastia. These cells might be constituents of normal breasts or, perhaps, present only in cases of increased breast cancer risk. PMID- 10465673 TI - Social interpersonal dimensions of the psychoses. AB - This paper looks at the patterns for the creation and social management of insanity and the involvements of those concerned. It describes an interaction model with people in a social reality of everyday living, built up of and defined by 'subjective' definitions of a situation. The psychiatric profession, involved in their conventional medical one-to-one confidential consultations with their patients, fails to be informed about the societal dimensions with two, three or more members as first described by Simmel (1902) (1). The differences are traditional, 'Two is company' and 'Three is a crowd'. If one of the possible two person relationships in a family develops some emotional change to define itself as 'Two is company', this may alter the emotional balance in the whole family and may lead to 'split minds' (schizo/phrenia) and involving relationships with and between other family members. The arrival of the first baby changes two to three and creates 'Our Family'. 'Two is company: but it is not our family.' 'Three is a crowd' but now, with three possible pairs, who is the odd one to be left out, or to push in or be pushed out? This is proposed as the interpersonal relationships substrate of the manic-(push in) or depressive (pushed out) psychoses in an older family generation. Both propositions are to be developed using probability theory to define the number of members, the corresponding numbers of their possible kinetic interpersonal relationships, their social dynamism probabilities, and potential outcomes involving modern non-linear mathematics. These patients are described as 'not themselves' or 'beside themselves'. Those who are themselves but described as neurotic or psychopathic will also be mentioned. PMID- 10465674 TI - Arterial stress from intraluminal pressure modified by tissue pressure offers a complete explanation for the distribution of atherosclerosis. AB - Pressure-induced arterial stress is probably the principle factor in the localization of atherosclerotic disease. The occurrence of lesions at the bifurcations and ostia of arteries seems to be caused by the concentration of stress at these locations. Other puzzles and paradoxes regarding the distribution of atherosclerosis can be explained by dynamic extra-arterial pressure (DEAP): fluctuations in tissue pressure that occur with movement, body position, and the beating of the heart. These tissue pressures, both above and below atmospheric pressure, change the transmural pressures in arteries and thereby either retard or accelerate atherogenesis. The DEAP hypothesis predicts that physical fitness increases extra-arterial tissue pressure, which protects important arteries from high transmural pressure. Deconditioning is expected to have the exact opposite effect, producing the equivalent of localized hypertension in the cerebral, internal carotid, coronary, abdominal aorta, and lower extremity arteries. PMID- 10465675 TI - Adoptive immunotherapy for cutaneous T-cell lymphomas. AB - The pathology of cutaneous T-cell lymphomas is predominantly in the skin. Allogeneic bone-marrow transplantation results in graft-versus-host disease involving the skin in many cases. It has been suggested that it would be interesting to see the effect of allotransplantation, even though no series has been reported so far. Adoptive immunotherapy with donor lymphocyte infusions has been increasingly used to treat post-transplant relapse and even prevention. The present hypothesis suggests that infusion of human leucocyte antigen (HLA) matched donor lymphocyte infusions would result in a graft-versus leukemia/lymphoma (GVL) effect in these disorders. Therefore, clinical trials are suggested. PMID- 10465676 TI - Short note: Thanks to the mammalian clones, the idea of totipotent somatic cells catches on (?). PMID- 10465677 TI - [New therapeutic attacks on cancer]. PMID- 10465678 TI - [Extra-thyroidal accompanying and resulting diseases]. PMID- 10465679 TI - Analysis of fibroblast proteins from patients with Duchenne muscular dystrophy by two-dimensional gel electrophoresis. AB - Duchenne muscular dystrophy (DMD), the most common and severe form of the muscular dystrophies, is an X-linked inborn error of metabolism with multiple tissue involvement. Although the major pathological changes are observed in skeletal muscle, abnormalities have also been detected in the heart, nervous system, red blood cells, lymphocytes and cultured skin fibroblasts. For many reasons, such as readily available tissue material, fewer secondary changes and the potential for prenatal diagnosis, cultured skin fibroblasts should be the tissue of choice to search for the primary defect. Several abnormalities have been reported in DMD fibroblasts, suggesting that the genetic abnormality is expressed in these cells. To search for potentially mutant protein(s) we have compared the protein composition of normal and DMD fibroblasts by two-dimensional gel electrophoresis and have now found one protein spot consistently missing in DMD cells. The nature of this protein and its relation to the DMD gene are unknown. PMID- 10465680 TI - Memantine is a clinically well tolerated N-methyl-D-aspartate (NMDA) receptor antagonist--a review of preclinical data. AB - N-methyl-D-aspartate (NMDA) receptor antagonists have therapeutic potential in numerous CNS disorders ranging from acute neurodegeneration (e.g. stroke and trauma), chronic neurodegeneration (e.g. Parkinson's disease, Alzheimer's disease, Huntington's disease, ALS) to symptomatic treatment (e.g. epilepsy, Parkinson's disease, drug dependence, depression, anxiety and chronic pain). However, many NMDA receptor antagonists also produce highly undesirable side effects at doses within their putative therapeutic range. This has unfortunately led to the conclusion that NMDA receptor antagonism is not a valid therapeutic approach. However, memantine is clearly an uncompetitive NMDA receptor antagonist at therapeutic concentrations achieved in the treatment of dementia and is essentially devoid of such side effects at doses within the therapeutic range. This has been attributed to memantine's moderate potency and associated rapid, strongly voltage-dependent blocking kinetics. The aim of this review is to summarise preclinical data on memantine supporting its mechanism of action and promising profile in animal models of chronic neurodegenerative diseases. The ultimate purpose is to provide evidence that it is indeed possible to develop clinically well tolerated NMDA receptor antagonists, a fact reflected in the recent interest of several pharmaceutical companies in developing compounds with similar properties to memantine. PMID- 10465681 TI - Alpha3beta4 subunit-containing nicotinic receptors dominate function in rat medial habenula neurons. AB - Regional-specific differences in nicotinic acetylcholine receptors (nAChRs) were examined using the whole-cell patch clamp technique in rat medial habenula (MHb) slices. The majority of cells in the ventral two thirds of the MHb responded robustly to local pressure application of nAChR agonists. Mean agonist potency profiles in the middle and ventral thirds of the MHb were similar: cytisine was the most potent agonist and DMPP the weakest, consistent with a significant contribution of the beta4 subunit to functional nAChRs in all areas of the MHb. In acutely isolated MHb neurons, the alpha3beta4-selective toxin alpha-CTx-AuIB (1 microM) reversibly blocked approximately 75% of the nicotine-induced currents, as expected for cells solely expressing alpha3beta4 nAChRs. However, the alpha3beta2-selective toxin, alpha-CTx-MII (100 nM), blocked a variable fraction (0-90%) of the MHb nicotinic response implying that beta2 subunits may contribute to some functional receptors. We suggest that the effects of alpha-CTx-MII may arise from interaction with alpha3beta2beta4 subunit-containing nAChRs. This idea is supported by the findings (1) that alpha-CTx-MII antagonizes receptors comprised of alpha3, beta2 and beta4 subunits in Xenopus oocytes, and (2) that a mutant alpha-CTx-MII toxin[H12A], which blocks alpha3beta2beta4 receptors but not alpha3beta2 or alpha3beta4 nAChRs, also reduces nicotinic currents in some MHb neurons. Overall these data imply that most functional nAChRs on MHb cells contain at least alpha3 and beta4 subunits, and that a variable subpopulation additionally contains the beta2 subunit. PMID- 10465682 TI - Functional integrity of green fluorescent protein conjugated glycine receptor channels. AB - The alpha subunit (alphaZ1) of the zebrafish glycine receptor (GlyR) has been N terminus fused with green fluorescent protein (GFP). We found that both pharmacological and electrophysiological properties of this chimeric alphaZ1-GFP are indistinguishable from those of the wild-type receptor when expressed in Xenopus oocytes and cell lines. The apparent affinities of this receptor for agonists (glycine, taurine and GABA), and the antagonist (strychnine) are unchanged, and single channel kinetics are not altered. In the same expression systems, alphaZ1-GFP was visualized using fluorescence microscopy. Fluorescence was distributed anisotropically across cellular membranes. In addition to the Golgi apparatus and endoplasmic reticulum, its presence was also detected on the plasmalemma, localized at discrete hot-spots which were identified as sites of high membrane turnover. Overall, the preservation in alphaZ1-GFPs of the wild type receptor functional properties makes it a promising new tool for further in situ investigations of GlyR expression, distribution and function. PMID- 10465683 TI - A simple method to transfer plasmid DNA into neuronal primary cultures: functional expression of the mGlu5 receptor in cerebellar granule cells. AB - We describe a method to transfer cDNA into neuronal primary cultures with a commercialised cationic lipid, Transfast. Cultures were transfected at a rate of about 5% with green fluorescent protein (GFP) cDNA. Comparing Transfast to other transfection reagents, we found this compound to be the most efficient. GFP transfected mouse cerebellar granule cells displayed normal whole-cell voltage sensitive and unitary big K+ channel currents. We also used this transfection method with success to transfer GFP cDNA into primary cultures of striatum and colliculus. Transfast was then used to cotransfect cultured cerebellar cells with GFP cDNA, in conjunction with cDNA coding for the metabotropic glutamate receptor type 5 (mGlu5 receptor). Ninety percent of the cells expressing GFP also expressed mGlu5 receptor. Though neurones were best transfected one day after plating, they still expressed both GFP and mGlu5 receptor proteins 2 weeks after plating, i.e. after full differentiation. A functional test of the expressed mGlu5 receptor was thus performed in GFP-transfected neurones. Stimulation of mGlu5 receptor induced single big K+ channel activity, as it was the case for the native mGlu1 receptor. This indicated that the transfected mGlu5 receptor plasmid was functionally expressed and that both mGlu1 and mGlu5 receptors may share common coupling mechanisms to big K+ channels in neurones. PMID- 10465684 TI - Regulation of synaptic transmission in the mossy fibre-granule cell pathway of rat cerebellum by metabotropic glutamate receptors. AB - The role of metabotropic glutamate receptors (mGluRs) in the mossy fibre-granule cell pathway in rat cerebellum was studied using slice preparations and electrophysiological techniques. Application of the group I selective agonist (S) 3,5-dihydroxyphenylglycine (DHPG) evoked, in a concentration-dependent manner (EC50 = 33 microM), a depolarising/hyperpolarising complex response from granule cells which was preferentially inhibited by the group I selective antagonist (S) 4-carboxyphenylglycine (4CPG). The group III selective agonist L-amino-4 phosphonobutyrate (AP4) evoked a hyperpolarising response (EC50 = 10 microM) which was inhibited by the group II/III selective antagonist (S)-alpha-methyl-4 phosphonophenylglycine (MPPG). The group II agonist (2S,2'R,3'R)-2-(2',3' dicarboxylcyclopropyl)glycine (DCG-IV) elicited no measurable voltage change. The amplitude of the synaptically-mediated mossy fibre response in granule cells was unaffected during application of AP4, was reduced by DHPG and was enhanced by DCG IV (EC50 = 80 nM). These effects were inhibited by the group selective antagonists 4CPG and (2S,1'S,2'S,3'R)-2-(2'-carboxy-3'-phenylcyclopropyl)glycine (PCCG-4), respectively. Further investigation using patch-clamp recording revealed that DCG-IV potently inhibited spontaneous GABAergic currents. We conclude that group I and III (but not group II) mGluRs are functionally expressed by granule cells, whereas unexpectedly group II or III mGluRs do not appear to be present presynaptically on mossy fibre terminals. Group II mGluRs are located on Golgi cell terminals; when activated these receptors cause disinhibition, a function which may be important for gating information transfer from the mossy fibres to the granule cells. PMID- 10465685 TI - The class I metabotropic glutamate receptor antagonist, AIDA, improves short-term and impairs long-term memory in a spatial task for rats. AB - Effects of the class I selective metabotropic glutamate receptor antagonist, 1 aminoindan-1,5-dicarboxylic acid (AIDA), on spatial procedural learning and episodic short-term memory of rats were investigated in an appetitively reinforced 3-choice delayed match-to-position task. First, an acute intraperitoneal injection of AIDA (2 mg/kg) was given 20 min before a single training session of 20 trials using repeated reward position in one alcove out of three. AIDA caused facilitated short-term acquisition within such a session compared to saline treated controls. Secondly, injections were given before each of ten sessions (48 h intervals) also using constant reward position. The results showed AIDA induced inhibition of procedural between-session acquisition. Finally, the use of reward positions in a non-repetitive but trial-specific version of the 3-choice test revealed a facilitating effect of AIDA on episodic short-term memory. PMID- 10465686 TI - Expression of heme oxygenase-1 mediated by non-NMDA and metabotropic receptors in glial cells: possible involvement of reactive oxygen species production and protein kinase C activation. AB - Heme oxygenase (HO) produces biliverdin and bilirubin which are physiological antioxidants and potent scavengers of oxygen radicals. Recently, we found that intracerebroventricular injection of kainic acid (KA) induced inducible HO (HO-1) predominantly in glial cells in the rat hippocampus in vivo. In this study, we examined the mechanism of HO-1 expression induced by agonists for glutamate receptors in cultured glial cells in vitro. The HO-1 protein level was significantly enhanced by several agonists for non-N-methyl-D-aspartate (non NMDA) receptors and metabotropic glutamate receptors (mGluR) such as KA, quisqualic acid (QA), (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propanoic acid (AMPA), and trans-(+/-)-1-amino-(1S,3R)-cyclopentanedicarboxylic acid (ACPD). Among these agonists, QA had the greatest potency. KA-induced HO-1 expression was inhibited by the non-NMDA antagonist NBQX. In addition, KA induced the marked production of reactive oxygen species (ROS), and KA-induced HO-1 expression was also inhibited by the antioxidants allopurinol and ascorbic acid. ACPD-induced HO-1 expression was inhibited by the mGluR antagonist MCPG and the protein kinase C (PKC) inhibitor calphostin C. These results suggest that induction of HO-1 expression by the activation of non-NMDA receptors is mediated by ROS production, and that expression induced by mGluR activation is mediated by PKC activation in rat glial cells. PMID- 10465687 TI - The role of systemic, spinal and supraspinal L-arginine-nitric oxide-cGMP pathway in thermal hyperalgesia caused by intrathecal injection of glutamate in mice. AB - The intrathecal (i.t.) administration of glutamate (10-100 nmol) caused dose related hyperalgesia (mean ED50 of 35 nmol) when assessed in the thermal behaviour model of nociception, the hot-plate test maintained at 50 degrees C. The i.p., i.t. or intracerebroventricular (i.c.v.) injection of the nitric oxide synthase inhibitors, L-NOARG and L-NAME, did not induce any detectable effect per se, but instead, produced dose-related inhibition of glutamate-induced hyperalgesia. D-NAME, the inactive enantiomer of L-NAME, had no effect. The i.c.v. or i.t. administration of L-NIO caused graded attenuation of glutamate induced hyperalgesia. L-arginine (3.4 mmol kg(-1), i.p.), but not D-arginine (3.4 mmol kg(-1), i.p.) significantly potentiated glutamate (10 nmol)-induced hyperalgesia, an action that was prevented by L-NOARG (137 nmol kg(-1)). The co injection of S-nitroso-N-acetyl-D,L-penicillamine (SNAP) (0.22 micromol) or 8 bromo-cGMP (22.5 nmol) with glutamate (10 nmol), via either i.t. or i.c.v. routes, also significantly enhanced glutamate-induced hyperalgesia. The guanylate cyclase inhibitors LY 83583 (0.1-1.0 nmol) or ODQ (30-300 pmol) co-administered with glutamate, dose-dependently antagonised the glutamate-induced hyperalgesia. Collectively, these results demonstrate that the i.t. injection of glutamate into the spinal cord of mice produces dose-related hyperalgesia an effect that was largely mediated by the L-arginine-nitric oxide-cGMP pathway from both spinal and supraspinal sites. PMID- 10465689 TI - Prenatal methylazoxymethanol acetate alters behavior and brain NGF levels in young rats: a possible correlation with the development of schizophrenia-like deficits. AB - It has been hypothesized that a deleterious key contribution to schizophrenia (SZ) development is a failure of migration and setting of young neurons into their appropriate cortical target sites, particularly in the entorhinal cortex (EC). To test this hypothesis in an animal model, we injected, in pregnant rats, on gestational day (GD) 9, or 10, or 11, or 12, the antimitotic compound methylazoxymethanolacetate (MAM) known to cause EC neuronal loss. We investigated whether or not EC disruption during prenatal development is able to affect behavior, including memory and learning, and brain nerve growth factor (NGF). Prenatally MAM treated young rats didn't display gross behavioral changes in social interaction, open-field and novel object investigation tests. By contrast, GD11 and GD12 MAM treated rats had a retardation in passive avoidance acquisition, while, in GD12 animals, pain sensitivity was reduced. GD12 animals also showed increased NGF in the EC and remaining cortex. MAM treated animals showed no changes in paw NGF or substance P levels suggesting that the altered nociceptive response is not related to local downregulation of these two molecules. The possibility that these behavioral and biochemical alterations might be associated with the onset of SZ is discussed. PMID- 10465688 TI - Blockade of Ba2+ current through human alpha1E channels by two steroid analogs, (+)-ACN and (+)-ECN. AB - Previous work suggests that different neuroactive steroids may exhibit some selectivity in their blocking effects on different high-voltage activated (HVA) Ca2+ currents. At least some of these effects appear to involve direct blocking actions on Ca2+ channels. Thus, direct investigation of the effects of various steroids on cloned Ca2+ channel variants may lead to the development of potent and selective small-molecular weight Ca2+ channel blockers. Here we examine the effects of two steroids on a cloned human alpha1E Ca2+ channel both with and without a beta3 subunit, when expressed in HEK293 cells. One compound, (+)-ACN, has been previously shown to block N-, Q-, and R-subtypes of HVA current without affecting L- and P-type current. The second compound, (+)-ECN, weakly blocks total HVA current in hippocampal neurons. (+)-ECN differs from (+)-ACN in lacking effects on GABA receptors, but shares with (+)-ACN an ability to partially inhibit T current in DRG neurons (Todorovic, S.M., Prakriya, M., Nakashima, Y.M. et al., 1998. Enantioselective blockade of T-type Ca2+ current in adult rat sensory neurons by a steroid lacking GABA-mimetic activity. Mol. Pharmacol. 54, 918-927). (+)-ACN can block 100% of Ba2+ current in HEK cells arising either from the alpha1E subunit (IC50 approximate to 10 microM) or the alpha1Ebeta3 combination (IC50 approximate to 5 microM), while (+)-ECN maximally blocks only about 80% of the alpha1E (10 microM) or alpha1Ebeta3 (16 microM) current. Blockade by (+)-ACN exhibits several differences from blockade by (+)-ECN. (+) ACN increases the apparent rate of onset of inactivation, particularly for the alpha1E variant, slows recovery from inactivation, and more profoundly shifts the voltage-dependence of current availability for both alpha1E and alpha1Ebeta3 variants than does (+)-ECN. Although the complexity of the normal inactivation kinetics of alpha1E variants makes interpretation of the (+)-ACN-induced kinetic alterations difficult, the results suggest that the two steroids are to some extent acting by distinct mechanisms, and perhaps at different sites. PMID- 10465691 TI - Drug-induced parkinsonism: cinnarizine and flunarizine are potent uncouplers of the vacuolar H+-ATPase in catecholamine storage vesicles. AB - Cinnarizine (1-diphenylmethyl-4-(3-phenyl-2-propenyl)piperazine) and its di fluorinated derivative flunarizine inhibit the MgATP-dependent generation of a transmembrane proton electrochemical gradient in chromaffin granule ghosts. The concentrations giving 50% inhibition (IC50) of the MgATP-dependent generation of the pH-gradient were 5.9+/-0.6 microM (n = 6) and 3.0+/-0.3 microM (n = 5) for cinnarizine and flunarizine, respectively. The IC50 values for inhibiting the generation of the membrane potential were even lower, i.e. 0.19+/-0.06 microM (n = 6) and 0.15+/-0.01 microM (n = 4) for cinnarizine and flunarizine, respectively. Cinnarizine (10 microM) also inhibited the energy-dependent vesicular uptake of [14C]-dopamine (50 microM) by 76%, i.e. from 2.1+/-0.9 to 0.5+/-0.6 nmol/mg protein/min (n = 5, P < 0.002). Cinnarizine (10 microM) increased the MgATPase activity of the granule ghosts by 47+/-26% (n = 4) compatible with an uncoupling of the vacuolar H+-ATPase activity. The IC50-values observed for the two compounds are in the same range as their reported therapeutic plasma concentrations in vivo, suggesting that cinnarizine and flunarizine may well inhibit proton pumping and catecholamine uptake in storage vesicles also in vivo. This mechanism of action may contribute to the drug induced parkinsonism seen as a side-effect of the two drugs. PMID- 10465690 TI - Further studies on the involvement of the arachidonic acid cascade in the acute dependence produced by mu, kappa and delta opioid agonists in isolated tissues. AB - The effects of phospholipase A2, cyclooxygenase-1, cyclooxygenase-2, and 5 lipoxygenase inhibitors on acute opiate withdrawal induced by selective mu, kappa and delta receptor agonists was investigated in vitro. After a 4 min in vitro exposure to D-Ala2-N-methyl-Phe-Gly5-ol)enkephalin (DAMGO; a highly selective mu agonist) and trans(+/-)-3,4-dichloro-N-methyl-N-(2(1pyrrolidynyl)-cyclohexyl)-+ ++benzeneacetamid (U50-488H; a highly selective K agonist) a strong contraction of the guinea pig isolated ileum was observed after the addition of naloxone. This effect was also observed when rabbit isolated jejunum was pretreated with deltorphin (a highly selective delta agonist). Mepacrine (a phospholipase A2 inhibitor), tolmetin (a selective cyclooxygenase-1 inhibitor) and meloxicam (a selective cyclooxygenase-2 inhibitor) treatment before or after DAMGO or U50-488H were able to both prevent and reverse the naloxone-induced contraction after exposure to the opioid agonists, in a concentration-dependent fashion. In addition, nordihydroguaiaretic acid (a 5-lipooxygenase inhibitor) was able to block the naloxone-induced contraction following exposure to DAMGO or U50-488H if injected either before or after the opioid agonist. In contrast, mepacrine, tolmetin, meloxicam and nordihydroguaiaretic acid did not affect the naloxone induced contraction after exposure to deltorphin. The results of the present study confirm and extend a previous study performed with morphine indicating that arachidonic acid and its metabolites (prostaglandins and leukotrienes) are involved in the development of opioid withdrawal induced by selective mu and kappa opioid agonists whereas no effects were observed on withdrawal induced by the selective delta opioid agonist deltorphin. PMID- 10465692 TI - Effect of reserpine on behavioural responses to agonists at 5-HT1A, 5-HT1B, 5 HT2A, and 5-HT2C receptor subtypes. AB - Rats were given a single dose of reserpine (5 mg/kg s.c.) and behavioural responses to agonists at 5-HT receptor subtypes compared with those of control animals 21 days later. The following effects of activating postsynaptic 5-HT1A receptors by the agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) were significantly increased: tail-flick, reciprocal forepaw treading, flat body posture. The hyperphagic effect of activating presynaptic 5-HT1A receptors by 8 OH-DPAT tended to increase and hypothermia on activating postsynaptic 5-HT1A sites tended to decrease. The hyperlocomotor effect of activating 5-HT1A sites also tended to decrease possibly as a result of a dependence of this response on the known depletion of catecholamines by reserpine. Head shakes on activating 5 HT2A receptors by 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) and two effects of activating 5-HT2C receptors by 1-(3-chlorophenyl) piperazine (mCPP) were significantly increased (hypophagia, anxiety) and a third effect, hypolocomotion tended to increase but hypophagia on activating postsynaptic 5 HT1B receptors by CP-94, 253 was significantly attenuated. The results are discussed with particular reference to altered 5-HT function in depression. PMID- 10465693 TI - Effects of repeated fluoxetine on anxiety-related behaviours, central serotonergic systems, and the corticotropic axis axis in SHR and WKY rats. AB - In keeping with the anxiolytic property of selective serotonin reuptake inhibitors (SSRIs) in humans, we have examined in the spontaneously hypertensive rat (SHR) and the Wistar-Kyoto (WKY) rat, which display low and high anxiety, respectively, some psychoneuroendocrine effects of a repeated treatment with the SSRI fluoxetine (5 or 10 mg/kg daily, for 3 weeks). Two days after the last injection, plasma levels of fluoxetine were not detectable whereas those of its metabolite, norfluoxetine, were present to similar extents in both strains. By means of the elevated plus-maze test (29-30 h after the 13th administration of fluoxetine) and an open field test (48 h after the last injection of fluoxetine), it was observed that fluoxetine pretreatment did not yield anxiolysis; hence, some, but not all, behaviours were indicative of anxiety and hypolocomotion (as assessed through principal component analyses and acute diazepam studies). In both strains, the 10 mg/kg dose of fluoxetine decreased hypothalamus 5-HT and 5 HIAA levels, and reduced midbrain and/or hippocampus [3H]citalopram binding at 5 HT transporters, but did not affect [3H]8-hydroxy-2-(di-N-propylamino)tetralin binding at hippocampal 5-HT1A receptors. However, the fluoxetine-elicited reduction in hippocampal 5-HT transporter binding was much more important in WKY than in SHR rats, this strain-dependent effect being associated in WKY rats with a reduction in cortical [3H]ketanserin binding at 5-HT2A receptors. Lastly, in WKY rats, repeated fluoxetine administration increased adrenal weights and the plasma corticosterone response to open field exposure, but did not affect the binding capacities of hippocampal mineralocorticoid and glucocorticoid receptors. These data show that key psychoneuroendocrine responses to repeated fluoxetine administration may be strain-dependent, and that repeated fluoxetine administration does not yield anxiolysis, as assessed by two standard tests of emotivity. PMID- 10465694 TI - (-)-Pindolol increases dialysate concentrations of dopamine and noradrenaline, but not serotonin, in the frontal cortex of freely-moving rats. AB - (-)-Pindolol (2.5-20.0 mg/kg, s.c.) markedly increased extracellular levels of dopamine (DA) and noradrenaline (NA) in single dialysate samples of the frontal cortex (FCX) of freely-moving rats. This action was specific inasmuch as serotonin (5-HT) levels were not significantly modified. In contrast, (-) propranolol (2.5) did not modify FCX dialysate levels of DA, NA (or 5-HT) alone and abolished the facilitatory influence of (-)-pindolol (10.0) upon levels of DA, though not NA. In contrast to (-)-propranolol, (-)-pindolol exerts, a facilitatory influence upon frontocortical dopaminergic and adrenergic, but not serotonergic, transmission. PMID- 10465695 TI - Neuroprotective mechanism of glial cell line-derived neurotrophic factor on dopamine neurons: role of antioxidation. AB - Recombinant human GDNF was infused into the rat striatum either acutely or subchronically. Its effects and its interactions with MPP+ on antioxidant enzyme activities were examined. Results indicated that acute GDNF infusion significantly increased glutathione peroxidase, superoxide dismutase and catalase activities. Subchronic GDNF treatment decreased the DA level and enhanced DA turnover. Pre-treatment with GDNF markedly protected DA neurons against MPP+ induced toxicity. These results suggest that GDNF protects DA neurons through its activation of the antioxidant enzyme systems. PMID- 10465696 TI - Systemic administration of cocaine, given alone or in combination with sensory stimuli, differentially affects L-arginine-nitric oxide metabolism in discrete regions of the brain of rat. AB - The effect of cocaine on brain regional metabolism of L-arginine to nitric oxide (NO) has been studied in rat by measuring the level of citrulline, the co-product of NO synthesis, using a HPLC based methodology. A single i.p. administration of 1 mg/kg cocaine, and a daily treatment for up to 5 consecutive days, failed to affect significantly citrulline content in the striatum, hippocampus and cortex. By contrast, in these regions of the brain a single or 5-day repeated higher dose of cocaine (10 mg/kg, i.p.) caused a significant increase in the co-product of NO synthesis and this has been abolished in a stereoselective fashion by L-NAME (10 mg/kg i.p. given 30 min before). Under cocaine high dose treatment, 1 h acoustic stimulation, which per se resulted ineffective, enhanced stimulant-induced increases in citrulline content seen in the striatum and abolished the increase of this amino acid observed in the hippocampus and cortex both after single or 5 day repeated injection of cocaine. In conclusion, these data demonstrate that cocaine stimulates the conversion of L-arginine to NO in the brain of rat and this is affected by concomitant exposure to acoustic stimulation. PMID- 10465697 TI - Influence of visual feedback on successive control mechanisms in upright quiet stance in humans assessed by fractional Brownian motion modelling. AB - An up-to-date way to model the centre of pressure (CP) trajectories may consist in using fractional Brownian motion (fBm). By doing so, one may note that standing still is in fact controlled by two separate and successive mechanisms. The point raised in this study concerns the nature of these control mechanisms and their level of interaction. Following this idea, visual feedback (VFB), which is known to affect postural control by significantly decreasing sway magnitudes, was used. Twelve healthy adults, instructed to stand as still as possible, were tested under this VFB protocol (via a PC screen). In order to model the CP trajectories as fBm, variograms (mean square distances, MSD, expressed as a function of increasing time intervals deltat) were bi-logarithmically plotted. The main visual effect of VFB on these variograms concerns longest latency scaling regimes which reveal less stochastic and consequently more accurate control (P < 0.05 and P < 0.01 for X and Y components, respectively). An increase in the MSD of the transition point, which corresponds to the switch between the two control mechanisms, is also noted (P < 0.05). Overall, evidence is provided from this data that long latency scaling regimes do operate through a feedback process. Interestingly, this improved determinism in feedback control in turn induces a similar effect on the control operating over the shortest deltat. Thus, by privileging a control strategy based on feedback mechanisms, VFB in turn would make the subjects quicker in their initial displacement in order to reach a position capable of initiating a feedback mechanism. PMID- 10465698 TI - The developmental expression and activity of peptidylarginine deiminase in the mouse. AB - We have measured the expression and activity of peptidylarginine deiminase (PAD, EC 3.5.3.15), the enzyme responsible for converting arginyl residues in proteins to citrullines, in normal mouse brain homogenate. PAD transcripts were detected as early as five days and were maximal at one month of age. The enzyme protein was also detected at 5 days in an antibody dependent assay and was maximal at 2 months of age, 1 month later than the maximum expression of transcripts. As expected, enzyme activity had a similar developmental profile to that of the enzyme protein. In isolated mouse brain compact myelin, the activity was highest at 15 days and fell rapidly to 15% of this level by 1-2 months. In the 'loose' myelin fraction (heavy myelin) it remained at the same high level form from 15 days to 8 months. The activity in compact myelin was about 15 times greater than the activity in brain homogenate, suggesting much of the enzyme was localized to myelin. PMID- 10465699 TI - 5-HT1B antagonists modulate clearance of extracellular serotonin in rat hippocampus. AB - In vivo chronoamperometry was used to determine the effects of locally applied antagonists of the serotonin-1B (5-HT1B) receptor (cyanopindolol, 5-HT-moduline and methiothepin) on the clearance of 5-HT in the CA3 region of the dorsal hippocampus and the corpus callosum (a brain region devoid of serotonin transporters and 5-HT1B receptors) of the rat. When 5-HT was pressure ejected into these regions, reproducible signals were detected. In the CA3 region, local application of 5-HT1B antagonists decreased the rate of clearance of the serotonin signal comparably to the selective 5-HT uptake inhibitor (SSRI), fluvoxamine. By contrast, in the corpus callosum, none of the drugs altered the 5 HT signal. One interpretation of these data is that 5-HT1B receptors can modulate the activity of the 5-HT transporter. PMID- 10465700 TI - Evidence that inhibitory motor neurons of the guinea-pig small intestine exhibit fast excitatory synaptic potentials mediated via P2X receptors. AB - Intracellular recordings were used to study the contribution of nicotinic and P2X receptors to synaptic transmission to morphologically identified myenteric neurons of guinea-pig ileum. Hexamethonium (100 microM) abolished fast excitatory synaptic potentials (EPSPs) in all orally projecting neurons, but fast EPSPs in anally projecting neurons were resistant to this antagonist. The non-cholinergic fast EPSPs were virtually abolished by suramin (100 microM). This suggests that P2X receptors are important in descending motility reflexes. However, suramin and hexamethonium together did not affect descending inhibitory reflexes when applied to the site of transmission between interneurons in this pathway. These data suggest that P2X receptors are not involved in transmission between descending interneurons, but may be important for transmission to inhibitory motor neurons. PMID- 10465702 TI - Muscarinic M2-receptors in rat thoracic dorsal root ganglia. AB - The occurrence and distribution of the muscarinic M2-receptor subtype (M2R) was investigated in rat thoracic dorsal root ganglia (DRG). Messenger RNA for M2R was demonstrated by RT-PCR in total RNA from DRG. Immunoreactivity to M2R-protein was localized to 26% of sensory neurons, the majority of them (85%) belonging to the size class of 25-40 microm in diameter. Double-labeling (immuno)histochemistry revealed that all M2R-immunoreactive neurons bind the lectin, I-B4, whereas they are generally devoid of substance P-immunoreactivity. These data show the presence of M2R on a subpopulation of presumably nociceptive primary afferent neurons, thereby extending previous pharmacological and electrophysiological studies that indicated a role of M2R and/or M4R in inhibition of calcium channel currents in rat sensory neurons (Wanke, E., Bianchi, L., Mantegazza, M., Guatteo, E., Macinelli, E. and Ferroni, A., Muscarinic regulation of Ca2+ currents in rat sensory neurons: channel and receptor types, dose-response relationships and cross-talk pathways. Eur. J. Neurosci., 6 (1994) 381-391). PMID- 10465701 TI - Fos expression in the trigeminocervical complex of the cat after stimulation of the superior sagittal sinus is reduced by L-NAME. AB - Primary neurovascular headaches, such as migraine and cluster headache probably involve activation of trigeminovascular pain structures projecting to the trigeminocervical complex of neurons in the caudal brain stem and upper cervical spinal cord. It has recently been demonstrated that blockade of the synthesis of nitric oxide (NO) by an NO synthesis inhibitor can abort acute migraine attacks and thus it is of interest to determine whether there is an influence of NO generation on trigeminocervical neurons. Cats were anaesthetised with alpha chloralose (60 mg/kg, i.t.). supplemental 20 mg/kg, intravenously (i.v.)) and halothane for surgery (0.5-3% by inhalation). A circular midline craniotomy was performed to isolate the superior sagittal sinus (SSS) for electrical stimulation (0.3 Hz, 150 V, 250 micros duration for 2 h). Two groups were compared, one stimulated after administration of vehicle and the other stimulated after administration of N(G)-nitro-L-arginine methylester (L-NAME: 100 mg/kg, i.v.). After stimulation of the SSS Fos immunoreactivity was observed in lamina I/IIo of the trigeminal nucleus caudalis and dorsal horns of C1 and C2 to a median total of 136 cells (range 122-146). After L-NAME treatment Fos expression was significantly reduced to 40 cells (24-54; P < 0.02). In conclusion, inhibition of NO synthesis L-NAME markedly reduces Fos expression in the trigeminocervical complex of the cat. These data taken together with the clinical observations of the effect of NO synthesis blockade in migraine suggest a role for NO generation in mediating nociceptive transmission in acute migraine. PMID- 10465703 TI - Specific whole-body shifts induced by frequency-modulated vibrations of human plantar soles. AB - This study sought to analyze the postural responses induced by separately or simultaneously vibrating with different frequencies the forefoot and rear foot zones of both soles in standing subjects. Stimulating each zone separately resulted in spatially oriented body tilts; their amplitude and velocity varied linearly according to the frequency, and their direction was always opposite to the plantar site vibrated. When the two zones were each co-stimulated at different frequencies, the parameters of the postural responses depended on the frequency difference. When this frequency difference was zero, no clearly oriented body tilts occurred. We concluded that the change in the relative pressures evoked by differently co-vibrating these zones gave rise to regulative postural adjustments able to cancel the simulated body deviation. PMID- 10465704 TI - Intracellular Ca2+ responses to nucleotides, peptides, amines, amino acids and prostaglandins in cultured pituicytes from adult rat neurohypophysis. AB - The present study aimed to investigate the reactivity of cultured pituicytes from adult neurohypophysis to various bioactive substances using Ca2+ indicator dye Fura-2. A transient increase of intracellular Ca2+ [Ca2+]i was observed when pituicytes were treated with nucleotides (ATP, ADP, UTP, and UDP) and amines (5 HT2 and alpha2-agonist). Treatment with peptides such as endothelin-1 (ET-1), endothelin-3 (ET-3), bradykinin (BK), vasopressin (AVP), and angiotensin II (Ang II) also induced [Ca2+]i increase in pituicytes. Prostaglandin E2 (PGE2) and F2alpha (PGF2alpha) increased [Ca2+]i, but amino acids of GABA, glutamate (Glu), and taurine had no effect. Serum-free culture condition augmented [Ca2+]i responses to ATP, Ang II and 5-HT within 24 h. These results indicate that pituicytes express many of receptors for neurotransmitters or neuromodulators. PMID- 10465705 TI - Central respiratory effects of substance P in neonatal mice: an in vitro study. AB - Experiments were performed on neonatal mice to know whether substance P (SP) modified the rhythm and the amplitude of the phrenic bursts generated in vitro in brainstem-cervical cord preparations. In OF1 and C3H neonatal preparations, SP or the tachykinin NK1 receptor agonist [Sar9,Met(O2)11] substance P both increased significantly phrenic burst amplitude (10(-7) M) but had no significant effect on respiratory rhythm unless used at concentrations 10 times larger. In neonates from the monoamine oxidase-A deficient transgenic Tg8 line, SP increased phrenic burst amplitude but had no effect on the respiratory rhythm at the tested concentrations. The role of SP in regulating neonatal respiratory activity is discussed on the basis of rat and mouse results. PMID- 10465706 TI - No genetic association between polymorphisms in the Tau gene and Alzheimer's disease in clinic or population based samples. AB - Mutations in the tau protein gene have recently been found to cause familial fronto-temporal dementia in a number of kindreds demonstrating linkage to chromosome 17. Given that tau pathology is a hallmark of Alzheimer's disease (AD), this raises the possibility that mutations in tau may also be associated with AD. We have investigated the allelic frequencies of polymorphisms in the Tau gene for a possible allelic distortion in Alzheimer's cases, which might suggest a conferred genetic risk. We have genotyped 65 community-based and 200 clinic based AD cases, and 142 community-based controls at the Tau exon 6 AflIII and BslI polymorphisms and find no independent association with risk for AD in these samples. Further analysis including APOE genotypes from the same samples demonstrated no interaction between either of these polymorphisms and APOE in conferring risk for AD. In addition, haplotype analysis across both sites revealed no difference in haplotype frequencies between cases and controls, nor any interaction with APOE. Therefore our data do not suggest any association between these variations in the Tau gene and Alzheimer's disease. PMID- 10465707 TI - Chronic treatment with the antidepressant amitriptyline decreases CRF-R1 receptor mRNA levels in the rat amygdala. AB - Using semi-quantitative in situ hybridization, corticotropin-releasing factor (CRF) and CRF receptor 1 (CRF-R1) mRNA levels were determined in the rat hypothalamus and amygdala after short-term (10 days) and chronic (4 weeks) treatment with the antidepressant amitriptyline. We found that chronic treatment with amitriptyline produced a significant decrease in CRF mRNA (to 33% of control) in the hypothalamic paraventricular nucleus (PVN). Short-term or chronic amitriptyline treatment had no effect on CRF-R1 mRNA levels in the PVN. However, after chronic treatment, there was a significant decrease of CRF-R1 mRNA levels in the lateral + basolateral (to 60% of control), and in the medial (to 70% of control) amygdala nuclei. These results suggest that the tricyclic antidepressant amitriptyline may exert part of its effects through modulation of hypothalamic CRF and of CRF-R1 gene expression in the amygdala. PMID- 10465708 TI - Blockade of GABA uptake potentiates GABA-induced depolarizations in adult mouse cortical slices. AB - The electrophysiological effects of GABA and the GABA uptake inhibitor, NO-711, were investigated on cortical slices prepared from adult audiogenic seizure-prone DBA/2 mice. GABA, perfused for 1 min, elicited depolarizing responses which had a mean duration of 80-100 s and were concentration-dependent (0.1-32 mM). NO-711 (25 microM), perfused for 15 min, produced depolarizations with a mean duration of 50-60 s and these persisted for 4-5 h. The responses to both compounds were blocked by the GABA(A) receptor antagonist, bicuculline. Pre-treatment of the slices with NO-711 potentiated the responses to GABA and moved the concentration response curve to the left. The EC50 to GABA following pre-treatment with NO-711 was 111+/-24 microM from a control value of 1.17+/-0.19 mM. These results demonstrate that in cortical slices GABA has a depolarizing action rather than the conventional one of hyperpolarization and that this response is potentiated by inhibition of GABA reuptake. PMID- 10465709 TI - Pyrrolidine dithiocarbamate prevents ethanol-induced elevation of [Ca2+]i in cultured canine cerebral vascular smooth muscle cells. AB - Pyrrolidine dithiocarbamate (PDTC) has been shown to block nuclear transcription factor (NF-kappaB) activation induced by a wide range of stimuli in different cell lines. NF-kappaB is a common element of the promoter region of inflammatory cytokines which can be stimulated by ethanol. Recently, we have shown that PDTC can ameliorate cerebrovascular damage, brain cortical damage, leukocyte adhesion and rolling, and stroke induced by ethanol. We, therefore, tested the effects of preincubation with PDTC on alcohol-induced changes in intracellular free calcium ions ([Ca2+]i) in cultured canine cerebral smooth muscle cells. These vascular cells, chronically treated with ethanol (10-100 mM) for 24 and 72 h, exhibited concentration-dependent rises in [Ca2+]i. PDTC (0.1 microM) itself failed to influence resting levels of [Ca2+]i in these vascular muscle cells. PDTC (0.1 microM) pretreatment, however, inhibited completely the elevations in [Ca2+]i induced by chronic ethanol (10-100 mM). The present results suggest that ethanol induced production of reactive oxygen species and elevation of [Ca2+]i in cultured canine cerebral vascular smooth muscle cells triggers induction of transcription factor NF-kappaB, which could play an important role in alcohol induced brain damage and stroke. PMID- 10465710 TI - Geomagnetic disturbances are associated with reduced nocturnal excretion of a melatonin metabolite in humans. AB - The effects of geomagnetic disturbances on urinary excretion of the melatonin metabolite, 6-hydroxymelatonin sulfate (6-OHMS), were studied in conjunction with 60 Hz magnetic field (MF) and ambient light exposure in 132 electric utility workers. Geomagnetic activity was assessed using a local (equivalent amplitude or A(K), Boulder, CO) and global (average antipodal or aa) index. Personal exposures to 60 Hz MFs and light were obtained using data-logging meters. The relationship between geomagnetic activity and 6-OHMS was assessed with adjustment for age, light exposure, and month of participation. Mean overnight 6-OHMS excretion was lower on days when the 36-h A(K) or aa values exceeded 30 nT. A greater reduction in 6-OHMS excretion was observed when increased geomagnetic activity was combined with elevated 60 Hz MF or reduced ambient light exposures. PMID- 10465712 TI - Noise stimulation decreases the concentration of norepinephrine in the rat cochlea. AB - The present study was designed to analyze, by using high performance liquid chromatography (HPLC), the effect of acoustic stimulation on the cochlear concentration of norepinephrine (NE). Independently of the rat strain (Long-Evans or Wistar strains), NE concentration decreased about 18% when animals were exposed to white noise (90 dB SPL for 1 h). The same decrease was observed in animals perfused by aortic pathway to remove the blood, indicating that this decrease corresponds exclusively to a neurophysiological process. In fact, these findings could indicate that noise stimulation is involved in the NE release from sympathetic fibers innervating the cochlea. This likely release of NE supports that sympathetic fibers play a functional role in cochleae exposed to noisy situations. PMID- 10465711 TI - Alpha-synuclein accumulates in Lewy bodies in Parkinson's disease and dementia with Lewy bodies but not in Alzheimer's disease beta-amyloid plaque cores. AB - A growing body of evidence suggests that the non-Abeta component of Alzheimer's disease amyloid precursor protein (NACP) or alpha-synuclein contributes to the neurodegenerative processes in Alzheimer's disease (AD), Parkinson's disease (PD) and dementia with Lewy bodies (DLB). In the present study antisera to the N terminus and the NAC domain of the alpha-synuclein protein were employed to elucidate the expression pattern in brains of patients with AD, PD, DLB and control specimen. Alpha-synuclein exhibited an overall punctuate expression profile compatible with a synaptic function. Interestingly, while Lewy bodies were strongly immunoreactive, none of the alpha-synuclein antisera revealed staining in mature beta-amyloid plaques in AD. These observations suggest that alpha-synuclein does not contribute to late neurodegenerative processes in AD brains. PMID- 10465713 TI - Nigral degeneration following striato-pallidal lesion in tissue type plasminogen activator deficient mice. AB - Tissue type plasminogen activator (tPA) has been suggested as a key factor in excitotoxic neuronal death in the hippocampus. Transneuronal degeneration of the substantia nigra pars reticulata (SNR) neurons after striato-pallidal lesions is attributable to excess excitatory glutamatergic inputs into the SNR following inhibitory GABAergic deafferentation and tPA may contribute to the mechanism of transneuronal degeneration of the SNR. To examine this possibility, we studied pathological changes in the SNR following striato-pallidal lesions produced by electrocoagulation in tPA-deficient mice. There was no difference in the degree of SNR degeneration, or in microglial activation and proliferation in the degenerating SNR of tPA-deficient and control mice. Our results indicate that tPA does not contribute to transneuronal degeneration in the SNR following striato pallidal lesions in mice. PMID- 10465714 TI - Nucleoside phosphorylation: a feasible step in the prebiotic pathway to RNA. AB - Plausible prebiotic conditions for the phosphorylation of nucleosides by inorganic phosphate were reported by Lohrmann and Orgel in 1971. This reaction was carried out on heated dry films and promoted by urea. The major products formed were nucleoside-2:3 cyclicPs;5-NMPs and other derivatives were also formed. Minor modifications of the Lohrmann and Orgel system have resulted in the preferential formation of 5-NMPs. In this modified system a 2-fold preference for phosphorylation of the 5-OH group over the 3(2)-OH group was observed and the formation of other derivatives was minimized. The small amounts of bis compounds that were formed in this system could be quantitatively removed by selective binding to the mineral hydroxylapatite at moderate ionic strengths. It was also discovered that under hydrolytic conditions there was a 3:1 preference for removal of phosphates attached to the 3-OH group over the 5-OH group. A recycling procedure for obtaining additonal 5-NMPs from bis compounds and 3-NMPs is proposed. PMID- 10465715 TI - Autocatalytic synthesis of oligoglycine in a simulated submarine hydrothermal system. AB - A flow reactor simulating a submarine hydrothermal system was constructed for examining the likelihood of oligopeptide synthesis from amino acids alone without recourse to any of condensing agents, templates or metallic ions. We demonstrated that when the initial reactant is glycine, the flow reactor can synthesize both di- and tri-glycine. The initial buildup of the yields of both the oligopeptides was found to be exponential with the elapse of time. The oligopeptide synthesis from glycine in the flow reactor could be autocatalytic. PMID- 10465716 TI - Enhanced photolysis of nucleic acid monomers by pyrophosphate in the simulated primitive soup. AB - In our simulation of the photochemistry of the primitive soup, it was found that yield of chromophore loss of some nucleic acid bases, nucleosides and nucleotides [NA] undergoing ultraviolet radiation by medium pressure mercury lamp [MPML] was enhanced by pyrophosphate and triphosphate whether O2 is present or not. The yield of chromophore loss of guanosine, uracil, 5 CMP, and the derivatives of adenine and thymine was observed to rise with the increase of molar concentration of pyrophosphate in N2-saturated systems. In air-saturated samples, increase in yield of chromophore loss was observed when the concentration of pyrophosphate reaches 5 x 10(-4) M, relative yield of chromophore loss (CL(rel)) coming to maximum in the range from 0.01 to 0.1 M, followed by a slight decline with the further increase of pyrophosphate concentration. The enhancement of photolysis of NA by pyrophosphate was due to the interaction between NA and pyrophosphate radicals photoionized by UV quanta of wavelength less than about 210 nm in the emission spectrum of a MPML. The relevance of this phenomenon to the origin of life has been discussed as well. PMID- 10465717 TI - The combination of salt induced peptide formation reaction and clay catalysis: a way to higher peptides under primitive earth conditions. AB - Two reactions with suggested prebiotic relevance for peptide evolution, the salt induced peptide formation reaction and the peptide chain elongation/stabilization on clay minerals have been combined in experimental series starting from dipeptides and dipeptide/amino acid mixtures. The results show that both reactions can take place simultaneously in the same reaction environment and that the presence of mineral catalysts favours the formation of higher oligopeptides. These findings lend further support to the relevance of these reactions for peptide evolution on the primitive earth. The detailed effects of the specific clay mineral depend both on the nature of the mineral and the reactants in solution. PMID- 10465719 TI - Enantioselective autocatalysis. V. The spontaneous resolution of tri-o-thymotide. AB - We have attempted to appraise experimentally the allegation that minute chemical and physical differences due to the parity violating energy difference (PVED) between enantiomers are, after suitable autocatalytic amplification, ultimately responsible for the homochirality of contemporary biomolecules. The autocatalytic amplification technique employed involved the spontaneous resolution under racemizing conditions (SRURC) of a conglomerate during crystallization, and the system studied was the known crystallization of P(+)- or M(-)-tri-o-thymotide (TOT) as its optically active inclusion compound (clathrate) with benzene. Our premise was that if a PVED effect were operative, there should be a strong and consistent bias favoring the crystallization of one enantiomer of the TOT-benzene clathrate. Repetitive preparations of the clathrate, however, yielded crystalline products showing random optical activity. These results thus afford no evidence whatsoever for stereoselective bias due to a PVED, and are in accord with earlier statistical studies demonstrating random SRURC in other conglomerate crystallizations, again indicating the inefficacy of PVEDs to promote a preferred chirality in such systems. PMID- 10465718 TI - Clay-nucleic acid complexes: characteristics and implications for the preservation of genetic material in primeval habitats. AB - The equilibrium adsorption of three nucleic acids: chromosomal DNA, supercoiled plasmid DNA, and 25S rRNA, on the clay minerals, montmorillonite (M) and kaolinite (K), were studied. Adsorption of the nucleic acid on the clays was rapid and maximal after 90 min of contact time. Chromosomal DNA was adsorbed to a greater extent than plasmid DNA and RNA, and the adsorption was also greater on M than on K. Adsorption isotherms were of the L type, and a plateau was reached with all the complexes, with the exception of chromosomal DNA adsorbed on M. To deterine where nucleic acids are adsorbed on clay minerals and the nature of the interaction, complexes were studied by X-ray diffraction (X-RD), electron microscopy, and Fourier transform infrared (FT-IR) spectroscopy. X-RD showed that nucleic acids did not penetrate the clay, indicating that the adsorption occurred primarily on the external surfaces of clay particles, as also suggested by electron microscopy observations. FT-IR spectra of clay-tightly bound nucleic acid complexes showed adsorption bands that indicate a variation of the nucleic acids status as a consequence of their adsorption on clay. Data obtained suggested that the formation of clay-nucleic acid complex could have an important role in the preservation of genetic material in primeval habitats. PMID- 10465720 TI - Extraterrestrial handedness: a reply. AB - Recent investigations of stable isotope ratios of amino acids from the Murchison meteorite have shown them to be of unambiguous extraterrestrial origin, and examinations of their enantiomeric compositions, where terrestrial contamination can be excluded, have found a consistent excess of L-enantiomers. One explanation for this observation has been the asymmetric photolysis of racemic extraterrestrial amino acids by circularly polarized light (CPL) in the synchrotron radiation from orbiting electrons around the pulsar remnants of supernovae. Mason (1997) has attempted to discredit this mechanism on the grounds that circular dichroism (CD) bands for optically active molecules alternate in sign and sum to zero over the entire spectrum, and hence enantioselective photochemical reactions cannot be induced by broad band CPL. We submit arguments disputing this conclusion and present reasons for expecting that broad band CPL synchrotron radiation would be quite capable of inducing asymmetric photolysis, particularly in aliphatic amino acids. PMID- 10465721 TI - Index of suspicion. Case 2. Brucellosis. PMID- 10465722 TI - Index of suspicion. Case 3. Hereditary spherocytosis. PMID- 10465723 TI - Markers for cardiac damage: current status and future trends. Proceedings of the 7th Bergmeyer Conference on Improving the Clinical Value of Laboratory Data. Tutzing, Germany, 1999. PMID- 10465724 TI - 38th Annual meeting of the Japanese Teratology Society. Nagoya, Aichi, Japan. July 22-24, 1998. Abstracts. PMID- 10465725 TI - Whence and whither cortex? PMID- 10465726 TI - Lipids and progressive renal disease: the cardio-renal link. PMID- 10465727 TI - L-carnitine use in hemodialyzed patients. PMID- 10465729 TI - Comparison of double-bag and Y-set disconnect systems in CAPD. PMID- 10465728 TI - Transferrin oversaturation. PMID- 10465730 TI - The house-dust mite: its biology and role in allergy. Proceedings of an international scientific workshop. Oslo, Norway, 4-7 September 1997. PMID- 10465732 TI - Proceedings of the 20th Symposium on Biotechnology for Fuels and Chemicals. Gatlinburg, Tennessee, USA. May 3-7, 1998. PMID- 10465731 TI - FENS Program for nutrition education in medical schools. Federation of European Nutrition Societies. PMID- 10465733 TI - Rhodopsin early receptor potential revisited. PMID- 10465734 TI - Modeling helix-turn-helix protein-induced DNA bending with knowledge-based distance restraints. AB - A crucial element of many gene functions is protein-induced DNA bending. Computer generated models of such bending have generally been derived by using a presumed bending angle for DNA. Here we describe a knowledge-based docking strategy for modeling the structure of bent DNA recognized by a major groove-inserting alpha helix of proteins with a helix-turn-helix (HTH) motif. The method encompasses a series of molecular mechanics and dynamics simulations and incorporates two experimentally derived distance restraints: one between the recognition helix and DNA, the other between respective sites of protein and DNA involved in chemical modification-enabled nuclease scissions. During simulation, a DNA initially placed at a distance was "steered" by these restraints to dock with the binding protein and bends. Three prototype systems of dimerized HTH DNA binding were examined: the catabolite gene activator protein (CAP), the phage 434 repressor (Rep), and the factor for inversion stimulation (Fis). For CAP-DNA and Rep-DNA, the root mean square differences between model and x-ray structures in nonhydrogen atoms of the DNA core domain were 2.5 A and 1.6 A, respectively. An experimental structure of Fis-DNA is not yet available, but the predicted asymmetrical bending and the bending angle agree with results from a recent biochemical analysis. PMID- 10465735 TI - Characterization of solute binding at human serum albumin site II and its geometry using a biochromatographic approach. AB - Chiral recognition mechanism relationships for binding at site II on human serum albumin (HSA) were investigated using D, L dansyl amino acids. Sodium phosphate salt was used as a solute-HSA interaction modifier. A new model was developed using a biochromatographic approach to describe the variation in the transfer equilibrium constant with the salt concentration, i.e., the nature of the interactions. The solute binding was divided into two salt concentration ranges c. For the low c values, below 0.03 M, the nonstereoselective interactions constituted the preponderant contribution to the variation in the solute binding with the salt concentration. For the high c values, above 0.03 M, the solute binding was governed by the hydrophobic effect and the stereoselective interactions. The different contributions implied in the binding process provided an estimation of both the surface charge density (sigma/F) and the surface area of the site II binding cavity accessible to solvent, which were found to be equal to around 10.10(-7) mol/m(2) and 2 nm(2). As well, the excess of sodium ions excluded by the solute transfer from the surface area of the pocket were about( 0.7) for dansyl norvaline and (-0.8) for dansyl tryptophan. PMID- 10465736 TI - Modeling electroporation in a single cell. I. Effects Of field strength and rest potential. AB - This study develops a model for a single cell electroporated by an external electric field and uses it to investigate the effects of shock strength and rest potential on the transmembrane potential V(m) and pore density N around the cell. As compared to the induced potential predicted by resistive-capacitive theory, the model of electroporation predicts a smaller magnitude of V(m) throughout the cell. Both V(m) and N are symmetric about the equator with the same value at both poles of the cell. Larger shocks do not increase the maximum magnitude of V(m) because more pores form to shunt the excess stimulus current across the membrane. In addition, the value of the rest potential does not affect V(m) around the cell because the electroporation current is several orders of magnitude larger than the ionic current that supports the rest potential. Once the field is removed, the shock-induced V(m) discharges within 2 micros, but the pores persist in the membrane for several seconds. Complete resealing to preshock conditions requires approximately 20 s. These results agree qualitatively and quantitatively with the experimental data reported by Kinosita and coworkers for unfertilized sea urchin eggs exposed to large electric fields. PMID- 10465737 TI - Modeling electroporation in a single cell. II. Effects Of ionic concentrations. AB - This study expands a previously developed model of a single cell electroporated by an external electric field by explicitly accounting for the ionic composition of the electroporation current. The previous model with non-specific electroporation current predicts that both the transmembrane potential V(m) and the pore density N are symmetric about the equator, with the same values at either end of the cell. The new, ion-specific case predicts that V(m) is symmetric and almost identical to the profile from the non-specific case, but N has a profound asymmetry with the pore density at the hyperpolarized end of the cell twice the value at the depolarized end. These modeling results agree with the experimentally observed preferential uptake of marker molecules at the hyperpolarized end of the cell as reported in the literature. This study also investigates the changes in intracellular ionic concentrations induced around an electroporated single cell. For all ion species, the concentrations near the membrane vary significantly, which may explain the electrical disturbances observed experimentally after large electric shocks are delivered to excitable cells and tissues. PMID- 10465738 TI - Long-term potentiation and depression induced by a stochastic conditioning of a model synapse. AB - Protracted presynaptic activity can induce long-term potentiation (LTP) or long term depression (LTD) of the synaptic strength. However, virtually all the experiments testing how LTP and LTD depend on the conditioning input are carried out with trains of stimuli at constant frequencies, whereas neurons in vivo most likely experience a stochastic variation of interstimulus intervals. We used a computational model of synaptic transmission to test if and to what extent the stochastic fluctuations of an input signal could alter the probability to change the state of a synapse. We found that, even if the mean stimulation frequency was maintained constant, the probability to induce LTD and LTP could be a function of the temporal variation of the input activity. This mechanism, which depends only on the statistical properties of the input and not on the onset of additional biochemical mechanisms, is not usually considered in the experiments, but it could have an important role to determine the amount of LTP/LTD induction in vivo. In response to a change in the distribution of the interstimulus intervals, as measured by the coefficient of variation, a synapse could be easily adapted to inputs that might require immediate attention, with a shift of the input thresholds required to elicit LTD or LTP, which are restored to their initial conditions as soon as the input pattern returns to the original temporal distribution. PMID- 10465739 TI - Model of intercellular calcium oscillations in hepatocytes: synchronization of heterogeneous cells. AB - Hepatocytes respond with repetitive cytosolic calcium spikes to stimulation by vasopressin and noradrenalin. In the intact liver, calcium oscillations occur in a synchronized fashion as periodic waves across whole liver lobules, but the mechanism of intercellular coupling remains unclear. Recently, it has been shown that individual hepatocytes can have very different intrinsic oscillation frequencies but become phase-locked when coupled by gap junctions. We investigate the gap junction hypothesis for intercellular synchronization by means of a mathematical model. It is shown that junctional calcium fluxes are effective in synchronizing calcium oscillations in coupled hepatocytes. An experimentally testable estimate is given for the junctional coupling coefficient required; it mainly depends on the degree of heterogeneity between cells. Intercellular synchronization by junctional calcium diffusion may occur also in other cell types exhibiting calcium-activated calcium release through InsP(3) receptors, if the gap junctional coupling is strong enough and the InsP(3) receptors are sufficiently sensitized by InsP(3). PMID- 10465740 TI - The osmotic migration of cells in a solute gradient. AB - The effect of a nonuniform solute concentration on the osmotic transport of water through the boundaries of a simple model cell is investigated. A system of two ordinary differential equations is derived for the motion of a single cell in the limit of a fast solute diffusion, and an analytic solution is obtained for one special case. A two-dimensional finite element model has been developed to simulate the more general case (finite diffusion rates, solute gradient induced by a solidification front). It is shown that the cell moves to regions of lower solute concentration due to the uneven flux of water through the cell boundaries. This mechanism has apparently not been discussed previously. The magnitude of this effect is small for red blood cells, the case in which all of the relevant parameters are known. We show, however, that it increases with cell size and membrane permeability, so this effect could be important for larger cells. The finite element model presented should also have other applications in the study of the response of cells to an osmotic stress and for the interaction of cells and solidification fronts. Such investigations are of major relevance for the optimization of cryopreservation processes. PMID- 10465741 TI - An analysis of the size selectivity of solute partitioning, diffusion, and permeation across lipid bilayers. AB - The lipid bilayers of cell membranes are primarily responsible for the low passive transport of nonelectrolytes across cell membranes, and for the pronounced size selectivity of such transport. The size selectivity of bilayer permeation has been hypothesized to originate from the hindered transport of solutes across the ordered-chain region. In this paper, we develop a theoretical description that provides analytical relationships between the permeation properties of the ordered-chain region of the lipid bilayer (partition and diffusion coefficients) and its structural properties, namely, lipid chain density, free area, and order parameter. Emphasis is placed on calculating the size selectivity of solute partitioning, diffusion, and overall permeability across the ordered-chain region of the lipid bilayer. The size selectivity of solute partitioning is evaluated using scaled-particle theory, which calculates the reversible work required to create a cavity to incorporate a spherical solute into the ordered-chain region of the lipid bilayer. Scaled-particle theory is also used to calculate the work required to create a diffusion path for solutes in the interfacial region of the lipid bilayer. The predicted size dependence of the bilayer permeability is comparable to that observed experimentally. The dependence of solute partition and diffusion coefficients on the bilayer structural parameters is also discussed. PMID- 10465742 TI - Molecular dynamics simulations of the complex between human U1A protein and hairpin II of U1 small nuclear RNA and of free RNA in solution. AB - RNA-protein interactions are essential to a wide range of biological processes. In this paper, a 0.6-ns molecular dynamics simulation of the sequence-specific interaction of human U1A protein with hairpin II of U1 snRNA in solution, together with a 1.2-ns simulation of the free RNA hairpin, is reported. Compared to the findings in the x-ray structure of the complex, most of the interactions remained stable. The nucleotide U8, one of the seven conserved nucleotides AUUGCAC in the loop region, was unusually flexible during the simulation, leading to a loss of direct contacts with the protein, in contrast to the situation in the x-ray structure. Instead the sugar-phosphate backbone of nucleotide C15 was found to form several interactions with the protein. Compared to the NMR structure of U1A protein complexed with the 3'-untranslated region of its own pre mRNA, the protein core kept the same conformation, and in the two RNA molecules the conserved AUUGCAC of the loop and the closest CG base pair were located in very similar positions and orientations, and underwent very similar interactions with the protein. Therefore, a common sequence-specific interaction mechanism was suggested for the two RNA substrates to bind to the U1A protein. Conformational analysis of the RNA hairpin showed that the conformational changes of the RNA primarily occurred in the loop region, which is just involved in the sites of binding to the protein and in agreement with experimental observation. Both the loop and stem of the RNA became more ordered upon binding to the protein. It was also demonstrated that the molecular dynamics method could be successfully used to simulate the dynamical behavior of a large RNA-protein complex in aqueous solution, thus opening a path for the exploration of the complex biological processes involving RNA at a molecular level. PMID- 10465744 TI - A polarization model overcoming the geometric restrictions of the laplace solution for spheroidal cells: obtaining new equations for field-induced forces and transmembrane potential. AB - We present a new model for a variety of electric polarization effects on oblate and prolate homogeneous and single-shell spheroids. For homogeneous spheroids the model is identical to the Laplace model. For single-shell spheres of cell-like geometry the calculated difference of the induced dipole moments is in the thousandths range. To solve Laplace's equation for nonspherical single-shell objects it is necessary to assume a confocal shell, which results in different cell membrane properties in the pole and equator regions, respectively. Our alternative model addresses this drawback. It assumes that the disturbance of the external field due to polarization may project into the medium to a characteristic distance, the influential radius. This parameter is related to the axis ratio of the spheroid over the depolarizing factors and allows us to determine the geometry for a finite resistor-capacitor model. From this model the potential at the spheroid's surface is obtained and, consequently, the local field inside a homogeneous spheroid is determined. In the single-shell case, this is the effective local field of an equivalent homogeneous spheroid. Finally, integration over the volume yields the frequency-dependent induced dipole moment. The resistor-capacitor approach allowed us to find simple equations for the critical and characteristic frequencies, force plateaus and peak heights of deformation, dielectrophoresis and electrorotation for homogeneous and single shell spheroids, and a more generalized equation for the induced transmembrane potential of spheroidal cells. PMID- 10465743 TI - A kinetic molecular model of the reversible unfolding and refolding of titin under force extension. AB - Molecular elasticity is a physicomechanical property that is associated with a select number of polypeptides and proteins, such as the giant muscle protein, titin, and the extracellular matrix protein, tenascin. Both proteins have been the subject of atomic force microscopy (AFM), laser tweezer, and other in vitro methods for examining the effects of force extension on the globular (FNIII/Ig like) domains that comprise each protein. In this report we present a time dependent method for simulating AFM force extension and its effect on FNIII/Ig domain unfolding and refolding. This method treats the unfolding and refolding process as a standard three-state protein folding model (U right arrow over left arrow T right arrow over left arrow F, where U is the unfolded state, T is the transition or intermediate state, and F is the fully folded state), and integrates this approach within the wormlike chain (WLC) concept. We simulated the effect of AFM tip extension on a hypothetical titin molecule comprised of 30 globular domains (Ig or FNIII) and 25% Pro-Glu-Val-Lys (PEVK) content, and analyzed the unfolding and refolding processes as a function of AFM tip extension, extension rate, and variation in PEVK content. In general, we find that the use of a three-state protein-folding kinetic-based model and the implicit inclusion of PEVK domains can accurately reproduce the experimental force-extension curves observed for both titin and tenascin proteins. Furthermore, our simulation data indicate that PEVK domains exhibit extensibility behavior, assist in the unfolding and refolding of FNIII/Ig domains in the titin molecule, and act as a force "buffer" for the FNIII/Ig domains, particularly at low and moderate extension forces. PMID- 10465745 TI - The fractal structure of glycogen: A clever solution to optimize cell metabolism. AB - Fractal objects are complex structures built with a simple procedure involving very little information. This has an obvious interest for living beings, because they are splendid examples of optimization to achieve the most efficient structure for a number of goals by means of the most economic way. The lung alveolar structure, the capillary network, and the structure of several parts of higher plant organization, such as ears, spikes, umbels, etc., are supposed to be fractals, and, in fact, mathematical functions based on fractal geometry algorithms can be developed to simulate them. However, the statement that a given biological structure is fractal should imply that the iterative process of its construction has a real biological meaning, i.e., that its construction in nature is achieved by means of a single genetic, enzymatic, or biophysical mechanism successively repeated; thus, such an iterative process should not be just an abstract mathematical tool to reproduce that object. This property has not been proven at present for any biological structure, because the mechanisms that build the objects mentioned above are unknown in detail. In this work, we present results that show that the glycogen molecule could be the first known real biological fractal structure. PMID- 10465747 TI - Role of individual surface charges of voltage-gated K channels. AB - Fixed charges on the extracellular surface of voltage-gated ion channels influence the gating. In previous studies of cloned voltage-gated K channels, we found evidence that the functional surface charges are located on the peptide loop between the fifth transmembrane segment and the pore region (the S5-P loop). In the present study, we determine the role of individual charges of the S5-P loop by correlating primary structure with experimentally calculated surface potentials of the previously investigated channels. The results suggest that contributions to the surface potential at the voltage sensor of the different residues varies in an oscillating pattern, with the first residue of the N terminal end of the S5-P loop, an absolutely conserved glutamate, contributing most. An analysis yields estimates of the distance between the residues and the voltage sensor, the first N-terminal residue being located at a distance of 5-6 A. To explain the results, a structural hypothesis, comprising an alpha-helical N terminal end of the S5-P loop, is presented. PMID- 10465746 TI - Time-resolved rhodopsin activation currents in a unicellular expression system. AB - The early receptor current (ERC) is the charge redistribution occurring in plasma membrane rhodopsin during light activation of photoreceptors. Both the molecular mechanism of the ERC and its relationship to rhodopsin conformational activation are unknown. To investigate whether the ERC could be a time-resolved assay of rhodopsin structure-function relationships, the distinct sensitivity of modern electrophysiological tools was employed to test for flash-activated ERC signals in cells stably expressing normal human rod opsin after regeneration with 11-cis retinal. ERCs are similar in waveform and kinetics to those found in photoreceptors. The action spectrum of the major R(2) charge motion is consistent with a rhodopsin photopigment. The R(1) phase is not kinetically resolvable and the R(2) phase, which overlaps metarhodopsin-II formation, has a rapid risetime and complex multiexponential decay. These experiments demonstrate, for the first time, kinetically resolved electrical state transitions during activation of expressed visual pigment in a unicellular environment (single or fused giant cells) containing only 6 x 10(6)-8 x 10(7) molecules of rhodopsin. This method improves measurement sensitivity 7 to 8 orders of magnitude compared to other time-resolved techniques applied to rhodopsin to study the role particular amino acids play in conformational activation and the forces that govern those transitions. PMID- 10465748 TI - 1H-NMR and circular dichroism spectroscopic studies on changes in secondary structures of the sodium channel inactivation gate peptides as caused by the pentapeptide KIFMK. AB - The pentapeptide KIFMK, which contains three clustered hydrophobic amino acid residues of isoleucine, phenylalanine, and methionine (IFM) in the sodium channel inactivation gate on the cytoplasmic linker between domains III and IV (III-IV linker), is known to restore fast inactivation to the mutant sodium channels having a defective inactivation gate or to accelerate the inactivation of the wild-type sodium channels. To investigate the docking site of KIFMK and to clarify the mechanisms for restoring the fast inactivation, we have studied the interactions between KIFMK and the fragment peptide in the III-IV linker GGQDIFMTEEQK (MP-1A; G1484-K1495 in rat brain IIA) by one- and two-dimensional (1)H-NMR and circular dichroism (CD) spectroscopies. KIFMK was found to increase the helical content of MP-1A in 80% trifluoroethanol (TFE) solution by approximately 11%. A pentapeptide, KIFMT, which can restore inactivation but less effectively than KIFMK, also increased the helical content of MP-1A, but to a lesser extent ( approximately 6%) than did KIFMK. In contrast, KDIFMTK, which is ineffective in restoring inactivation, decreased the helical content ( approximately -4%). Furthermore, we studied the interactions between KIFMK and modified peptides from MP-1A, that is, MP-1NA (D1487N), MP-1QEA (E1492Q), or MP 1EQA (E1493Q). The KIFMK was found to increase the helical content of MP-1EQA to an extent nearly identical to that of MP-1A, whereas it was found to decrease those of MP-1NA and MP-1QEA. These findings mean that KIFMK, by allowing each of the Lys residues to interact with D1487 and E1492, respectively, stabilized the helical structure of the III-IV linker around the IFM residues. This helix stabilizing effect of KIFMK on the III-IV linker may restore and/or accelerate fast inactivation to the sodium channels having a defective inactivation gate or to wild-type sodium channels. PMID- 10465749 TI - Molecular dissection of transjunctional voltage dependence in the connexin-32 and connexin-43 junctions. AB - Most gap junction channels are sensitive to the voltage difference between the two cellular interiors, termed the transjunctional voltage (V(j)). In several junctions, the conductance transitions induced by V(j) show more than one kinetic component. To elucidate the structural basis of the fast and slow components that characterize the V(j )dependence of connexin-32 (Cx32) and connexin-43 (Cx43) junctions, we created deletions of both connexins, where most of the carboxy terminal (CT) domain was removed. The wild-type and "tailless" mutants were expressed in paired Xenopus oocytes, and the macroscopic gating properties were analyzed using the dual voltage clamp technique. Truncation of the CT domain of Cx32 and Cx43 abolished the fast mechanism of conductance transitions and induced novel gating properties largely attributable to the slow mechanism of gating. The formation of hybrid junctions comprising wild-type and truncated hemichannels allowed us to infer that the fast and slow components of gating reside in each hemichannel and that both gates close at a negative V(j) on the cytoplasmic side. Thus we conclude that the two kinetic components of V(j)-sensitive conductance are a result of the action of two different gating mechanisms. They constitute separate structures in the Cx32 and Cx43 molecules, the CT domain being an integral part of fast V(j) gating. PMID- 10465750 TI - Structural determinants of slow inactivation in human cardiac and skeletal muscle sodium channels. AB - Skeletal and heart muscle excitability is based upon the pool of available sodium channels as determined by both fast and slow inactivation. Slow inactivation in hH1 sodium channels significantly differs from slow inactivation in hSkM1. The beta(1)-subunit modulates fast inactivation in human skeletal sodium channels (hSkM1) but has little effect on fast inactivation in human cardiac sodium channels (hH1). The role of the beta(1)-subunit in sodium channel slow inactivation is still unknown. We used the macropatch technique on Xenopus oocytes to study hSkM1 and hH1 slow inactivation with and without beta(1)-subunit coexpression. Our results indicate that the beta(1)-subunit is partly responsible for differences in steady-state slow inactivation between hSkM1 and hH1 channels. We also studied a sodium channel chimera, in which P-loops from each domain in hSkM1 sodium channels were replaced with corresponding regions from hH1. Our results show that these chimeras exhibit hH1-like properties of steady-state slow inactivation. These data suggest that P-loops are structural determinants of sodium channel slow inactivation, and that the beta(1)-subunit modulates slow inactivation in hSkM1 but not hH1. Changes in slow inactivation time constants in sodium channels coexpressed with the beta(1)-subunit indicate possible interactions among the beta(1)-subunit, P-loops, and the slow inactivation gate in sodium channels. PMID- 10465751 TI - Contribution of ryanodine receptor type 3 to Ca(2+) sparks in embryonic mouse skeletal muscle. AB - The kinetic behavior of Ca(2+) sparks in knockout mice lacking a specific ryanodine receptor (RyR) isoform should provide molecular information on function and assembly of clusters of RyRs. We examined resting Ca(2+) sparks in RyR type 3 null intercostal myotubes from embryonic day 18 (E18) mice and compared them to Ca(2+) sparks in wild-type (wt) mice of the same age and to Ca(2+) sparks in fast twitch muscle cells from the foot of wt adult mice. Sparks from RyR type 3-null embryonic cells (368 events) were significantly smaller, briefer, and had a faster time to peak than sparks from wt cells (280 events) of the same age. Sparks in adult cells (220 events) were infrequent, yet they were highly reproducible with population means smaller than those in embryonic RyR type 3 null cells but similar to those reported in adult amphibian skeletal muscle fibers. Three-dimensional representations of the spark peak intensity (DeltaF/Fo) vs. full width at half-maximal intensity (FWHM) vs. full duration at half-maximal intensity (FTHM) showed that wt embryonic sparks were considerably more variable in size and kinetics than sparks in adult muscle. In all cases, tetracaine (0.2 mM) abolished Ca(2+) spark activity, whereas caffeine (0.1 mM) lengthened the spark duration in wt embryonic and adult cells but not in RyR type 3-null cells. These results confirmed that sparks arose from RyRs. The low caffeine sensitivity of RyR type 3-null cells is entirely consistent with observations by other investigators. There are three conclusions from this study: i) RyR type-1 engages in Ca(2+) spark activity in the absence of other RyR isoforms in RyR type 3-null myotubes; ii) Ca(2+) sparks with parameters similar to those reported in adult amphibian skeletal muscle can be detected, albeit at a low frequency, in adult mammalian skeletal muscle cells; and iii) a major contributor to the unusually large Ca(2+) sparks observed in normal (wt) embryonic muscle is RyR type 3. To explain the reduction in the size of sparks in adult compared to embryonic skeletal muscle, we suggest that in embryonic muscle, RyR type 1 and RyR type 3 channels co-contribute to Ca(2+) release during the same spark and that Ca(2+) sparks undergo a maturation process which involves a decrease in RyR type 3. PMID- 10465753 TI - Determining the activation time course of synaptic AMPA receptors from openings of colocalized NMDA receptors. AB - Excitatory postsynaptic currents (EPSCs) in most mammalian central neurons have a fast alpha-amino-3-hydroxy-5-methyl-4-isoazole-proprionic acid (AMPA) receptor mediated component, lasting a few milliseconds, and a slow N-methyl-D-aspartic acid (NMDA)-receptor-mediated component, lasting hundreds of milliseconds. The time course of the AMPA phase is crucial in the integrative function of neurons, but measuring it accurately is often confounded by cable filtering between the recording electrode and the synapse. We describe a method for recovering the AMPA phase of individual EPSCs by determining the impulse response of the cable filter from single NMDA channel transitions in the slow tails of the same EPSC, then deconvolving the measured AMPA current. Using simulations, we show that filtering of an AMPA conductance transient in a voltage-clamped dendrite behaves in an almost perfectly linear fashion. Expressions are derived for the time course of single channel transitions and the AMPA phase filtered through a voltage-clamped cable or a single exponential filter, using a kinetic model for AMPA receptor activation. Fitting these expressions to experimental records directly estimates the underlying kinetics of the AMPA phase. Example measurements of spontaneous EPSCs in cultured nonpyramidal rat cortical neurons yielded rising time constants of 0.2-0.8 ms, and decay time constants of 1.3-2 ms at 23-25 degrees C. PMID- 10465752 TI - Roles of electric field and fiber structure in cardiac electric stimulation. AB - This study investigated roles of the variation of extracellular voltage gradient (VG) over space and cardiac fibers in production of transmembrane voltage changes (DeltaV(m)) during shocks. Eleven isolated rabbit hearts were arterially perfused with solution containing V(m)-sensitive fluorescent dye (di-4-ANEPPS). The epicardium received shocks from symmetrical or asymmetrical electrodes to produce nominally uniform or nonuniform VGs. Extracellular electric field and DeltaV(m) produced by shocks in the absolute refractory period were measured with electrodes and a laser scanner and were simulated with a bidomain computer model that incorporated the anterior left ventricular epicardial fiber field. Measurements and simulations showed that fibers distorted extracellular voltages and influenced the DeltaV(m). For both uniform and nonuniform shocks, DeltaV(m) depended primarily on second spatial derivatives of extracellular voltages, whereas the VGs played a smaller role. Thus, 1) fiber structure influences the extracellular electric field and the distribution of DeltaV(m); 2) the DeltaV(m) depend on second spatial derivatives of extracellular voltage. PMID- 10465755 TI - Polymer-cushioned bilayers. I. A structural study of various preparation methods using neutron reflectometry. AB - This neutron reflectometry study evaluates the structures resulting from different methods of preparing polymer-cushioned lipid bilayers. Four different techniques to deposit a dimyristoylphosphatidylcholine (DMPC) bilayer onto a polyethylenimine (PEI)-coated quartz substrate were examined: 1) vesicle adsorption onto a previously dried polymer layer; 2) vesicle adsorption onto a bare substrate, followed by polymer adsorption; and 3, 4) Langmuir-Blodgett vertical deposition of a lipid monolayer spread over a polymer-containing subphase to form a polymer-supported lipid monolayer, followed by formation of the outer lipid monolayer by either 3) horizontal deposition of the lipid monolayer or 4) vesicle adsorption. We show that the initial conditions of the polymer layer are a critical factor for the successful formation of our desired structure, i.e., a continuous bilayer atop a hydrated PEI layer. Our desired structure was found for all methods investigated except the horizontal deposition. The interaction forces between these polymer-supported bilayers are investigated in a separate paper (Wong, J. Y., C. K. Park, M. Seitz, and J. Israelachvili. 1999. Biophys. J. 77:1458-1468), which indicate that the presence of the polymer cushion significantly alters the interaction potential. These polymer-supported bilayers could serve as model systems for the study of transmembrane proteins under conditions more closely mimicking real cellular membrane environments. PMID- 10465754 TI - The interaction of bioactive peptides with an immobilized phosphatidylcholine monolayer. AB - The interaction of three bioactive peptides, bombesin, beta-endorphin, and glucagon with a phosphatidylcholine monolayer that was immobilized to porous silica particles and packed into a stainless steel column cartridge, has been studied using dynamic elution techniques. This immobilized lipid monolayer provides a biophysical model system with which to study the binding of peptides to a lipid membrane. In particular, the influence of temperature and methanol concentration on the affinity of each peptide for the immobilized lipid surface was assessed. For all test peptides, nonlinear retention plots were observed at all temperatures that contrasted sharply with the simple linear plots observed for the small unstructured control molecules N-acetyltryptophanamide and diphenylalanine. An analysis of the thermodynamics of the interaction of peptides with the immobilized monolayer was also carried out. The results revealed that while the peptides interacted with the monolayer predominantly through hydrophobic interactions, the relative contribution of DeltaH(assoc)(O) and DeltaS(assoc)(O) to the overall free energy of association was dependent on the temperature and methanol concentration. In particular, it was evident that under most conditions, the binding of the peptides to the immobilized lipid monolayer was enthalpy-driven, i.e., mediated by nonclassical hydrophobic interactions. Significant band-broadening and asymmetric and split peaks were also observed for bombesin, beta-endorphin, and glucagon at different temperatures and methanol concentrations. These changes in affinity and peak shape are consistent with the formation of multiple conformational species during the interaction of these peptides with the lipid monolayer. In addition, the binding behavior of the three test peptides on an n-octylsilica surface that lacked the phospho headgroups of the phospholipid was significantly different from that observed with the immobilized phosphatidylcholine surface, indicating a specificity of interaction between the peptides and the lipid surface. Overall, these experimental results demonstrate that the biomimetic phosphatidylcholine monolayer provides a stable and sensitive system with which to explore the molecular mechanism of peptide conformational changes during membrane interactions. PMID- 10465757 TI - Interactions of pulmonary surfactant protein A with phospholipid monolayers change with pH. AB - The interaction of pulmonary surfactant protein A (SP-A) labeled with Texas Red (TR-SP-A) with monolayers containing zwitterionic and acidic phospholipids has been studied at pH 7.4 and 4.5 using epifluorescence microscopy. At pH 7.4, TR-SP A expanded the pi-A isotherms of film of dipalmitoylphosphatidylcholine (DPPC). It interacted at high concentration at the edges of condensed-expanded phase domains, and distributed evenly at lower concentration into the fluid phase with increasing pressure. At pH 4.5, TR-SP-A expanded DPPC monolayers to a slightly lower extent than at pH 7.4. It interacted primarily at the phase boundaries but it did not distribute into the fluid phase with increasing pressure. Films of DPPC/dipalmitoylphosphatidylglycerol (DPPG) 7:3 mol/mol were somewhat expanded by TR-SP-A at pH 7.4. The protein was distributed in aggregates only at the condensed-expanded phase boundaries at all surface pressures. At pH 4.5 TR-SP-A caused no expansion of the pi-A isotherm of DPPC/DPPG, but its fluorescence was relatively homogeneously distributed throughout the expanded phase at all pressures studied. These observations can be explained by a combination of factors including the preference for SP-A aggregates to enter monolayers at packing dislocations and their disaggregation in the presence of lipid under increasing pressure, together with the influence of pH on the aggregation state of SP-A and the interaction of SP-A with zwitterionic and acidic lipid. PMID- 10465756 TI - Polymer-cushioned bilayers. II. An investigation of interaction forces and fusion using the surface forces apparatus. AB - We have created phospholipid bilayers supported on soft polymer "cushions" which act as deformable substrates (see accompanying paper, Wong, J. Y., J. Majewski, M. Seitz, C. K. Park, J. N. Israelachvili, and G. S. Smith. 1999. Biophys. J. 77:1445-1457). In contrast to "solid-supported" membranes, such "soft-supported" membranes can exhibit more natural (higher) fluidity. Our bilayer system was constructed by adsorption of small unilamellar dimyristoylphosphatidylcholine (DMPC) vesicles onto polyethylenimine (PEI)-supported Langmuir-Blodgett lipid monolayers on mica. We used the surface forces apparatus (SFA) to investigate the long-range forces, adhesion, and fusion of two DMPC bilayers both above and below their main transition temperature (T(m) approximately 24 degrees C). Above T(m), hemi-fusion activation pressures of apposing bilayers were considerably smaller than for solid-supported bilayers, e.g., directly supported on mica. After separation, the bilayers naturally re-formed after short healing times. Also, for the first time, complete fusion of two fluid (liquid crystalline) phospholipid bilayers was observed in the SFA. Below T(m) (gel state), very high pressures were needed for hemi-fusion and the healing process became very slow. The presence of the polymer cushion significantly alters the interaction potential, e.g., long-range forces as well as fusion pressures, when compared to solid supported systems. These fluid model membranes should allow the future study of integral membrane proteins under more physiological conditions. PMID- 10465758 TI - Interaction of phloretin with lipid monolayers: relationship between structural changes and dipole potential change. AB - Phloretin is known to adsorb to lipid surfaces and alters the dipole potential of lipid monolayers and bilayers. Its adsorption to biological and artificial membranes results in a change of the membrane permeability for a variety of charged and neutral compounds. In this respect phloretin represents a model substance to study the effect of dipole potentials on membrane permeability. In this investigation we studied the interaction of phloretin with monolayers formed of different lipids in the liquid-expanded and the condensed state. Phloretin integrated into the monolayers as a function of the aqueous concentration of its neutral form, indicated by an increase of the surface pressure in the presence of phloretin. Simultaneous recording of the surface potential of the monolayers allowed us to correlate the degree of phloretin integration and the phloretin induced dipole potential change. Increasing the surface pressure decreased the phloretin-induced shift of the isotherms, but did not influence the phloretin induced surface potential change. This means that phloretin adsorption to the lipid surface can occur without affecting the lipid packing. The surface potential effect of phloretin is accompanied by a change of the lipid dipole moment vector dependent on the lipid packing. This means that the relation between the surface potential change and the lipid packing cannot be described by a static model alone. Taking into account the deviations of the surface potential change versus molecular area isotherms of the experimental data to the theoretically predicted course, we propose a model that relates the area change to the dipole moment in a dynamic manner. By using this model the experimental data can be described much better than with a static model. PMID- 10465759 TI - Ceramides modulate protein kinase C activity and perturb the structure of Phosphatidylcholine/Phosphatidylserine bilayers. AB - We studied the effects of natural ceramide and a series of ceramide analogs with different acyl chain lengths on the activity of rat brain protein kinase C (PKC) and on the structure of bovine liver phosphatidylcholine (BLPC)/dipalmitoylphosphatidylcholine (DPPC)/dipalmitoylphosphatidylserine (DPPS) (3:1:1 molar ratio) bilayers using (2)H-NMR and specific enzymatic assays in the absence or presence of 7.5 mol % diolein (DO). Only a slight activation of PKC was observed upon addition of the short-chain ceramide analogs (C(2)-, C(6)-, or C(8)-ceramide); natural ceramide or C(16)-ceramide had no effect. In the presence of 7.5 mol % DO, natural ceramide and C(16)-ceramide analog slightly attenuated DO-enhanced PKC activity. (2)H-NMR results demonstrated that natural ceramide and C(16)-ceramide induced lateral phase separation of gel-like and liquid crystalline domains in the bilayers; however, this type of membrane perturbation has no direct effect on PKC activity. The addition of both short-chain ceramide analogs and DO had a synergistic effect in activating PKC, with maximum activity observed with 20 mol % C(6)-ceramide and 15 mol % DO. Further increases in C(6) ceramide and/or DO concentrations led to decreased PKC activity. A detailed (2)H NMR investigation of the combined effects of C(6)-ceramide and DO on lipid bilayer structure showed a synergistic effect of these two reagents to increase membrane tendency to adopt nonbilayer structures, resulting in the actual presence of such structures in samples exceeding 20 mol % ceramide and 15 mol % DO. Thus, the increased tendency to form nonbilayer lipid phases correlates with increased PKC activity, whereas the actual presence of such phases reduced the activity of the enzyme. Moreover, the results show that short-chain ceramide analogs, widely used to study cellular effects of ceramide, have biological effects that are not exhibited by natural ceramide. PMID- 10465760 TI - Comparison of the biophysical properties of racemic and d-erythro-N-acyl sphingomyelins. AB - In this study stereochemically pure d-erythro-sphingomyelins (SMs) with either 16:0 or 18:1(cisDelta9) as the N-linked acyl-chain were synthesized. Our purpose was to examine the properties of these sphingomyelins and acyl-chain matched racemic (d-erythro/l-threo) sphingomyelins in model membranes. Liquid-expanded d erythro-N-16:0-SM in monolayers was observed to pack more densely than the corresponding racemic sphingomyelin. Cholesterol desorption to beta-cyclodextrin was significantly slower from d-erythro-N-16:0-SM monolayers than from racemic N 16:0-SM monolayers. Significantly more condensed domains were seen in cholesterol/d-erythro-N-16:0-SM monolayers than in the corresponding racemic mixed monolayers, when [7-nitrobenz-2-oxa-1, 3-diazol-4-yl]phosphatidylcholine was used as a probe in monolayer fluorescence microscopy. With monolayers of N 18:1-SMs, both the lateral packing densities (sphingomyelin monolayers) and the rates of cholesterol desorption (mixed cholesterol/sphingomyelin monolayers) was found to be similar for d-erythro and racemic sphingomyelins. The phase transition temperature and enthalpy of d-erythro-N-16:0-SM in bilayer membranes were slightly higher compared with the corresponding racemic sphingomyelin (41.1 degrees C and 8.4 +/- 0.4 kJ/mol, and 39.9 degrees C and 7.2 +/- 0.2 kJ/mol, respectively). Finally, d-erythro-sphingomyelins in monolayers (both N-16:0 and N 18:1 species) were not as easily degraded at 37 degrees C by sphingomyelinase (Staphylococcus aureus) as the corresponding racemic sphingomyelins. We conclude that racemic sphingomyelins differ significantly in their biophysical properties from the physiologically relevant d-erythro sphingomyelins. PMID- 10465761 TI - Condensed complexes of cholesterol and phospholipids. AB - Mixtures of dihydrocholesterol and phospholipids form immiscible liquids in monolayer membranes at the air-water interface under specified conditions of temperature and 2-dimensional pressure. In recent work it has been discovered that a number of these mixtures exhibit two upper miscibility critical points. Pairs of upper critical points can be accounted for by a theoretical model that implies the cooperative formation of molecular complexes of dihydrocholesterol and phospholipid molecules. These complexes are calculated to be present in the membranes both above and below the critical points. Below the critical points the complexes form a separate phase, whereas above the critical points the complexes are completely miscible with the other lipid components. The cooperativity of complex formation prompts the use of the terminology condensed complex. PMID- 10465762 TI - Dynein-ADP as a force-generating intermediate revealed by a rapid reactivation of flagellar axoneme. AB - Fragmented flagellar axonemes of sand dollar spermatozoa were reactivated by rapid photolysis of caged ATP. After a time lag of 10 ms, axonemes treated with protease started sliding disintegration. Axonemes without protease digestion started nanometer-scale high-frequency oscillation after a similar time lag. Force development in the sliding disintegration was measured with a flexible glass needle and its time course was corresponded well to that of the dynein-ADP intermediate production estimated using kinetic rates previously reported. However, with a high concentration ( approximately 80 microM) of vanadate, which binds to the dynein-ADP intermediate and forms a stable complex of dynein-ADP vanadate, the time course of force development in sliding disintegration was not affected at all. In the case of high frequency oscillation, the time lag to start the oscillation, the initial amplitude, and the initial frequency were not affected by vanadate, though the oscillation once started was damped more quickly at higher concentrations of vanadate. These results suggest that during the initial turnover of ATP hydrolysis, force generation of dynein is not blocked by vanadate. A vanadate-insensitive dynein-ADP is postulated as a force-generating intermediate. PMID- 10465763 TI - Shape, size, and distribution of Ca(2+) release units and couplons in skeletal and cardiac muscles. AB - Excitation contraction (e-c) coupling in skeletal and cardiac muscles involves an interaction between specialized junctional domains of the sarcoplasmic reticulum (SR) and of exterior membranes (either surface membrane or transverse (T) tubules). This interaction occurs at special structures named calcium release units (CRUs). CRUs contain two proteins essential to e-c coupling: dihydropyridine receptors (DHPRs), L-type Ca(2+) channels of exterior membranes; and ryanodine receptors (RyRs), the Ca(2+) release channels of the SR. Special CRUs in cardiac muscle are constituted by SR domains bearing RyRs that are not associated with exterior membranes (the corbular and extended junctional SR or EjSR). Functional groupings of RyRs and DHPRs within calcium release units have been named couplons, and the term is also loosely applied to the EjSR of cardiac muscle. Knowledge of the structure, geometry, and disposition of couplons is essential to understand the mechanism of Ca(2+) release during muscle activation. This paper presents a compilation of quantitative data on couplons in a variety of skeletal and cardiac muscles, which is useful in modeling calcium release events, both macroscopic and microscopic ("sparks"). PMID- 10465765 TI - Spermine-induced aggregation of DNA, nucleosome, and chromatin. AB - We have analyzed the conditions of aggregation or precipitation of DNA in four different states: double-stranded DNA (dsDNA), single-stranded DNA (ssDNA), mononucleosome core particles (NCP), and H1-depleted chromatin fragments (ChF) in the presence of the multivalent cation spermine (4+). In an intermediate regime of DNA concentration, these conditions are identical for the four states. This result demonstrates that the mechanism involved is general from flexible chains to rigid rods and quasi-colloidal states. It is dominated by local electrostatic attractions that are considered, for instance, by the "ion-bridging" model. The onset of precipitation does not require the electroneutrality of the DNA chains. Above a given spermine concentration dsDNA aggregates remain neutral, whereas NCP aggregates turn positively charged. The difference is thought to originate from the extension of the positively charged proteic tails of the NCP. This suggests that local fluctuations of polyamine concentrations can induce either positively or negatively charged chromatin domains. PMID- 10465764 TI - Tropomyosin modulates pH dependence of isometric tension. AB - We investigated the effect of pH on isometric tension in actin filament reconstituted and thin filament-reconstituted bovine cardiac muscle fibers in the pH range of 6.0-7.4. Thin filament was reconstituted from purified G-actin with either bovine cardiac tropomyosin (Tm) or rabbit skeletal Tm in conjunction with cardiac or skeletal troponin (Tn). Results showed that isometric tension decreased linearly with a decrease in pH. The slope of the pH-tension relation, DeltaF/DeltapH (Deltarelative tension/Deltaunit pH), was 0.28 and 0.44 in control cardiac fibers and skeletal fibers, respectively. In actin filament-reconstituted fibers without regulatory proteins, DeltaF/DeltapH was 0.62, namely larger than that in cardiac or skeletal fibers. When reconstituted with cardiac Tm-Tn complex (nTm), DeltaF/DeltapH recovered to 0.32, close to the value obtained in control cardiac fibers. When reconstituted with skeletal nTm, DeltaF/DeltapH recovered to 0.48, close to the value for control skeletal fibers. To determine whether Tm or Tn is responsible for the inhibitory effects of nTm on the tension decrease caused by reduced pH, thin filament was reconstituted with cardiac Tm and skeletal Tn, or with skeletal Tm and cardiac Tn. When cardiac Tm was used, pH dependence of isometric tension coincided with that of control cardiac fibers. When skeletal Tm was used, the pH dependence coincided with that of control skeletal fibers. Furthermore, closely similar results were obtained in fibers reconstituted with actin and either cardiac or skeletal Tm without Tn. These results demonstrate that Tm but not Tn modulates the pH dependence of active tension. PMID- 10465766 TI - Structural effects of cobalt-amine compounds on DNA condensation. AB - Light scattering and electron microscopy have been used to investigate the structural effects of the trivalent complexes hexaammine cobalt (III) chloride (Cohex), tris(ethylenediamine) cobalt(III) chloride (Coen), and cobalt(III) sepulchrate chloride (Cosep) on DNA condensation. These cobalt-amine compounds have similar ligand coordination geometries but differ slightly in size. Their hydrophobicity is in the order Cosep > Coen > Cohex, according to the numbers of methylene groups in these ligands. All of these compounds effectively precipitate DNA at high concentrations; but despite a lower surface charge density, Cosep condenses DNA twice as effectively as Coen or Cohex. UV and CD measurements of the supernatants of cobalt-amine/DNA solutions reveal a preferential binding of Delta-Coen over Lambda-Coen to the precipitated DNA, but there is no chiral selectivity for Cosep. Competition experiments show that the binding strengths of these three cobalt-amine compounds to aggregated DNA are comparable. A charge neutralization of 88-90% is required for DNA condensation. Our data indicate that 1) electrostatic interaction is the main driving force for binding of multivalent cations to DNA; 2) DNA condensation is dependent on the structure of the condensing agent; and 3) the hydration pattern or polarization of water molecules on the surface of condensing agents plays an important role in DNA condensation and chiral recognition. PMID- 10465768 TI - Conformational changes of bacteriorhodopsin along the proton-conduction chain as studied with (13)C NMR of [3-(13)C]Ala-labeled protein: arg(82) may function as an information mediator. AB - We have recorded (13)C NMR spectra of [3-(13)C]Ala-labeled wild-type bacteriorhodopsin (bR) and its mutants at Arg(82), Asp(85), Glu(194), and Glu(204) along the extracellular proton transfer chain. The upfield and downfield displacements of the single carbon signals of Ala(196) (in the F-G loop) and Ala(126) (at the extracellular end of helix D), respectively, revealed conformational differences in E194D, E194Q, and E204Q from the wild type. The same kind of conformational change at Ala(126) was noted also in the Y83F mutant, which lacks the van der Waals contact between Tyr(83) and Ala(126) present in the wild type. The absence of a negative charge at Asp(85) in the site-directed mutant D85N induced global conformational changes, as manifested in displacements or suppression of peaks from the transmembrane helices, cytoplasmic loops, etc., as well as the local changes at Ala(126) and Ala(196) seen in the other mutants. Unexpectedly, no conformational change at Ala(126) was observed in R82Q (even though Asp(85) is protonated at pH 6) or in D85N/R82Q. The changes induced in the Ala(126) signal when Asp(85) is uncharged could be interpreted therefore in terms of displacement of the positive charge of Arg(82) toward Tyr(83), where Ala(126) is located. It is possible that disruption of the proton transfer chain after protonation of Asp(85) in the photocycle could cause the same kind of conformational change we detect at Ala(196) and Ala(126). If so, the latter change would be also the result of rearrangement of the side chain of Arg(82). PMID- 10465767 TI - A molecular model for RecA-promoted strand exchange via parallel triple-stranded helices. AB - A number of studies have concluded that strand exchange between a RecA-complexed DNA single strand and a homologous DNA duplex occurs via a single-strand invasion of the minor groove of the duplex. Using molecular modeling, we have previously demonstrated the possibility of forming a parallel triple helix in which the single strand interacts with the intact duplex in the minor groove, via novel base interactions (Bertucat et al., J. Biomol. Struct. Dynam. 16:535-546). This triplex is stabilized by the stretching and unwinding imposed by RecA. In the present study, we show that the bases within this triplex are appropriately placed to undergo strand exchange. Strand exchange is found to be exothermic and to result in a triple helix in which the new single strand occupies the major groove. This structure, which can be equated to so-called R-form DNA, can be further stabilized by compression and rewinding. We are consequently able to propose a detailed, atomic-scale model of RecA-promoted strand exchange. This model, which is supported by a variety of experimental data, suggests that the role of RecA is principally to prepare the single strand for its future interactions, to guide a minor groove attack on duplex DNA, and to stabilize the resulting, stretched triplex, which intrinsically favors strand exchange. We also discuss how this mechanism can incorporate homologous recognition. PMID- 10465769 TI - The effect of temperature and solution pH on the nucleation of tetragonal lysozyme crystals. AB - Part of the challenge of macromolecular crystal growth for structure determination is obtaining crystals with a volume suitable for x-ray analysis. In this respect an understanding of the effect of solution conditions on macromolecule nucleation rates is advantageous. This study investigated the effects of supersaturation, temperature, and pH on the nucleation rate of tetragonal lysozyme crystals. Batch crystallization plates were prepared at given solution concentrations and incubated at set temperatures over 1 week. The number of crystals per well with their size and axial ratios were recorded and correlated with solution conditions. Crystal numbers were found to increase with increasing supersaturation and temperature. The most significant variable, however, was pH; crystal numbers changed by two orders of magnitude over the pH range 4.0-5.2. Crystal size also varied with solution conditions, with the largest crystals obtained at pH 5.2. Having optimized the crystallization conditions, we prepared a batch of crystals under the same initial conditions, and 50 of these crystals were analyzed by x-ray diffraction techniques. The results indicate that even under the same crystallization conditions, a marked variation in crystal properties exists. PMID- 10465770 TI - vpu transmembrane peptide structure obtained by site-specific fourier transform infrared dichroism and global molecular dynamics searching. AB - The recently developed method of site-directed Fourier transform infrared dichroism for obtaining orientational constraints of oriented polymers is applied here to the transmembrane domain of the vpu protein from the human immunodeficiency virus type 1 (HIV-1). The infrared spectra of the 31-residue long vpu peptide reconstituted in lipid vesicles reveal a predominantly alpha helical structure. The infrared dichroism data of the (13)C-labeled peptide yielded a helix tilt beta = (6.5 +/- 1.7) degrees from the membrane normal. The rotational pitch angle omega, defined as zero for a residue located in the direction of the helix tilt, is omega = (283 +/- 11) degrees for the (13)C labels Val(13)/Val(20) and omega = (23 +/- 11) degrees for the (13)C labels Ala(14)/Val(21). A global molecular dynamics search protocol restraining the helix tilt to the experimental value was performed for oligomers of four, five, and six subunits. From 288 structures for each oligomer, a left-handed pentameric coiled coil was obtained, which best fits the experimental data. The structure reveals a pore occluded by Trp residues at the intracellular end of the transmembrane domain. PMID- 10465771 TI - Ice-binding surface of fish type III antifreeze. AB - We employed computational techniques, including molecular docking, energy minimization, and molecular dynamics simulation, to investigate the ice-binding surface of fish type III antifreeze protein (AFP). The putative ice-binding site was previously identified by mutagenesis, structural analysis, and flatness evaluation. Using a high-resolution x-ray structure of fish type III AFP as a model, we calculated the ice-binding interaction energy of 11 surface patches chosen to cover the entire surface of the protein. These various surface patches exhibit small but significantly different ice-binding interaction energies. For both the prism ice plane and an "ice" plane in which water O atoms are randomly positioned, our calculations show that a surface patch containing 14 residues (L19, V20, T18, S42, V41, Q9, P12, A16, M21, T15, Q44, I13, N14, K61) has the most favorable interaction energy and corresponds to the previously identified ice-binding site of type III AFP. Although in general agreement with the earlier studies, our results also suggest that the ice-binding site may be larger than the previously identified "core" cluster that includes mostly hydrophilic residues. The enlargement mainly results from the inclusion of peripheral hydrophobic residues and K61. PMID- 10465772 TI - Helix packing in polytopic membrane proteins: role of glycine in transmembrane helix association. AB - The nature and distribution of amino acids in the helix interfaces of four polytopic membrane proteins (cytochrome c oxidase, bacteriorhodopsin, the photosynthetic reaction center of Rhodobacter sphaeroides, and the potassium channel of Streptomyces lividans) are studied to address the role of glycine in transmembrane helix packing. In contrast to soluble proteins where glycine is a noted helix breaker, the backbone dihedral angles of glycine in transmembrane helices largely fall in the standard alpha-helical region of a Ramachandran plot. An analysis of helix packing reveals that glycine residues in the transmembrane region of these proteins are predominantly oriented toward helix-helix interfaces and have a high occurrence at helix crossing points. Moreover, packing voids are generally not formed at the position of glycine in folded protein structures. This suggests that transmembrane glycine residues mediate helix-helix interactions in polytopic membrane proteins in a fashion similar to that seen in oligomers of membrane proteins with single membrane-spanning helices. The picture that emerges is one where glycine residues serve as molecular notches for orienting multiple helices in a folded protein complex. PMID- 10465774 TI - Single-event analysis of the packaging of bacteriophage T7 DNA concatemers in vitro. AB - Bacteriophage T7 packages its double-stranded DNA genome in a preformed protein capsid (procapsid). The DNA substrate for packaging is a head-to-tail multimer (concatemer) of the mature 40-kilobase pair genome. Mature genomes are cleaved from the concatemer during packaging. In the present study, fluorescence microscopy is used to observe T7 concatemeric DNA packaging at the level of a single (microscopic) event. Metabolism-dependent cleavage to form several fragments is observed when T7 concatemers are incubated in an extract of T7 infected Escherichia coli (in vitro). The following observations indicate that the fragment-producing metabolic event is DNA packaging: 1) most fragments have the hydrodynamic radius (R(H)) of bacteriophage particles (+/-3%) when R(H) is determined by analysis of Brownian motion; 2) the fragments also have the fluorescence intensity (I) of bacteriophage particles (+/-6%); 3) as a fragment forms, a progressive decrease occurs in both R(H) and I. The decrease in I follows a pattern expected for intracapsid steric restriction of 4',6-diamidino-2 phenylindole (DAPI) binding to packaged DNA. The observed in vitro packaging of a concatemer's genomes always occurs in a synchronized cluster. Therefore, the following hypothesis is proposed: the observed packaging of concatemer-associated T7 genomes is cooperative. PMID- 10465773 TI - Cooperative folding units of escherichia coli tryptophan repressor. AB - A previously published computational procedure was used to identify cooperative folding units within tryptophan repressor. The theoretical results predict the existence of distinct stable substructures in the protein chain for the monomer and the dimer. The predictions were compared with experimental data on structure and folding of the repressor and its proteolytic fragments and show excellent agreement for the dimeric form of the protein. The results suggest that the monomer, the structure of which is currently unknown, is likely to have a structure different from the one it has within the context of the highly intertwined dimer. Application of this method to the repressor monomer represents an extension of the computations into the realm of evaluating hypothetical structures such as those produced by threading. PMID- 10465775 TI - The lifetime of insulin hexamers. AB - The kinetic stability of insulin hexamers containing two metal ions was investigated by means of hybridization experiments. Insulin was covalently labeled at the N(epsilon)-amino group of Lys(B29) by a fluorescence donor and acceptor group, respectively. The labels neither affect the tertiary structure nor interfere with self-association. Equimolar solutions of pure donor and acceptor insulin hexamers were mixed, and the hybridization was monitored by fluorescence resonance energy transfer. With the total insulin concentration remaining constant and the association/dissociation equilibria unperturbed, the subunit interchange between hexamers is an entropy-driven relaxation process that ends at statistical distribution of the labels over 16 types of hexamers differing by their composition. The analytical description of the interchange kinetics on the basis of a plausible model has yielded the first experimental values for the lifetime of the hexamers. The lifetime is reciprocal to the product of the concentration of the exchanged species and the interchange rate constant: tau = 1/(c. k). Measured for different concentrations, temperatures, metal ions, and ligand-dependent conformational states, the lifetime was found to cover a range from minutes for T(6) to days for R(6) hexamers. The approach can be used under an unlimited variety of conditions. The information it provides is of obvious relevance for the handling, storage, and pharmacokinetic properties of insulin preparations. PMID- 10465776 TI - Swelling studies on the cornea and sclera: the effects of pH and ionic strength. AB - The biophysical properties of the cornea and sclera depend on the precise maintenance of tissue hydration. We have studied the swelling of the tissues as a function of pH and ionic strength of the bathing medium, using an equilibration technique that prevents the loss of proteoglycans during swelling. Synchrotron x ray diffraction was used to measure the average intermolecular and interfibrillar spacings, the fibril diameters, and the collagen D-periodicity. We found that both tissues swelled least near pH 4, that higher hydrations were achieved at lower ionic strengths, and that sclera swelled about one-third as much as cornea under most conditions. In the corneal stroma, the interfibrillar spacing increased most with hydration at pH values near 7. Fibril diameters and D periodicity were independent of tissue hydration and pH at hydrations above 1. Intermolecular spacings in both tissues decreased as the ionic strength was increased, and there was a significant difference between cornea and sclera. Finally, we observed that corneas swollen near pH 7 transmitted significantly more light than those swollen at lower pH levels. The results indicate that the isoelectric points of both tissues are close to pH 4. The effects of ionic strength can be explained in terms of chloride binding within the tissues. The higher light transmission achieved in corneas swollen at neutral pH may be related to the fact that the interfibrillar fluid is more evenly distributed under these conditions. PMID- 10465777 TI - Analysis of the mechanisms of mitochondrial NADH regulation in cardiac trabeculae. AB - We have previously shown that increased cardiac work initially caused a rapid Ca(2+)-independent fall of mitochondrial [NADH] ([NADH](m)) to a minimum level, and this was followed by a slow Ca(2+)-dependent recovery toward control level (Brandes and Bers, Biophys. J. 71:1024-1035, 1996; Brandes and Bers, Circ. Res. 80:82-87, 1997). The purpose of this study is to improve our understanding of the factors that control [NADH](m) during increased work. [NADH](m) was monitored using fluorescence spectroscopy in intact rat trabeculae isolated from the right ventricular wall. Work was increased by increasing sarcomere length, pacing frequency, external [Ca(2+)], or by decreased temperature. The results were: 1) The initial fall of [NADH](m) during increased pacing frequency depends independently on increased myofilament work and on increased Ca(2+)-transport ATPase activity. 2) The [NADH](m) recovery process depends on average cytosolic [Ca(2+)] (Av[Ca(2+)](c)), but not on absolute work level. 3) The initial fall of [NADH](m) and the [NADH](m) recovery are similar whether increased work is associated with low frequency and high Ca(2+)-transient amplitude or vice versa (at the same myofilament work level and Av[Ca(2+)](c)). 4) The mechanisms associated with the smaller fall and recovery of [NADH](m) at 37 degrees C versus 27 degrees C, may be explained by lowered Av[Ca(2+)](c) and myofilament work. The NADH control mechanisms that operate at lower temperature are thus qualitatively similar at more physiological temperatures. PMID- 10465778 TI - Blistering of langmuir-blodgett bilayers containing anionic phospholipids as observed by atomic force microscopy. AB - Asymmetric bilayers of different phospholipid compositions have been prepared by the Langmuir-Blodgett (L-B) method, and imaged by atomic force microscopy (AFM). Such bilayers can function as a model for biological membranes. The first leaflet consisted of zwitterionic phospholipids phosphatidylcholine (PC) or phosphatidylethanolamine (PE). The second leaflet consisted of the anionic phospholipid phosphatidylglycerol (PG), in either the condensed or liquid phase or, for comparison, of PC. Different bilayers showed different morphology. In all bilayers defects in the form of holes were present. In some bilayers with a first leaflet consisting of PC, polygonal line-shaped defects were observed, whereas when the first leaflet consisted of PE, mainly round defects were seen. Not only the shape, but also the amount of defects varied, depending on the condition and the composition of the second leaflet. In most of the PG-containing systems the defects were surrounded by elevations, which reversibly disappeared in the presence of divalent cations. This is the first time that such elevations have been observed on phospholipid bilayers. We propose that they are induced by phospholipid exchange between the two leaflets around the defects, leading to the presence of negatively charged phospholipids in the first leaflet. Because the substrate is also negatively charged, the bilayer around the edges is repelled and lifted up. Since it was found that the elevations are indeed detached from the substrate, we refer to this effect as bilayer blistering. PMID- 10465779 TI - The second derivative electronic absorption spectrum of cytochrome c oxidase in the Soret region. AB - The electronic absorption spectrum of solubilized beef heart cytochrome c oxidase was analyzed in the 400-500 nm region to identify the origin of doublet features appearing in the second derivative spectrum associated with ferrocytochrome a. This doublet, centered near 22,600 cm(-1), was observed in the direct absorption spectrum of the a(2+)a(3)(3+).HCOO(-) form of the enzyme at cryogenic temperatures. Since evidence for this doublet at room temperature is obtained only on the basis of the second derivative spectrum, a novel mathematical approach was developed to analyze the resolving power of second derivative spectroscopy as a function of parameterization of spectral data. Within the mathematical limits defined for resolving spectral features, it was demonstrated that the integrated intensity of the doublet feature near 450 nm associated with ferrocytochrome a is independent of the ligand and oxidation state of cytochrome a(3). Furthermore, the doublet features, also observed in cytochrome c oxidase from Paracoccus denitrificans, were similarly associated with the heme A component and were correspondingly independent of the ligand and oxidation state of the heme A(3) chromophore. The doublet features are attributed to lifting of the degeneracy of the x and y polarized components of the B state of the heme A chromophore associated with the Soret transition. PMID- 10465780 TI - Electron-nuclear double resonance and hyperfine sublevel correlation spectroscopic studies of flavodoxin mutants from Anabaena sp. PCC 7119. AB - The influence of the amino acid residues surrounding the flavin ring in the flavodoxin of the cyanobacterium Anabaena PCC 7119 on the electron spin density distribution of the flavin semiquinone was examined in mutants of the key residues Trp(57) and Tyr(94) at the FMN binding site. Neutral semiquinone radicals of the proteins were obtained by photoreduction and examined by electron nuclear double resonance (ENDOR) and hyperfine sublevel correlation (HYSCORE) spectroscopies. Significant differences in electron density distribution were observed in the flavodoxin mutants Trp(57) --> Ala and Tyr(94) --> Ala. The results indicate that the presence of a bulky residue (either aromatic or aliphatic) at position 57, as compared with an alanine, decreases the electron spin density in the nuclei of the benzene flavin ring, whereas an aromatic residue at position 94 increases the electron spin density at positions N(5) and C(6) of the flavin ring. The influence of the FMN ribityl and phosphate on the flavin semiquinone was determined by reconstituting apoflavodoxin samples with riboflavin and with lumiflavin. The coupling parameters of the different nuclei of the isoalloxazine group, as detected by ENDOR and HYSCORE, were very similar to those of the native flavodoxin. This indicates that the protein conformation around the flavin ring and the electron density distribution in the semiquinone form are not influenced by the phosphate and the ribityl of FMN. PMID- 10465781 TI - The actin-based nanomachine at the leading edge of migrating cells. AB - Two fundamental parameters of the highly dynamic, ultrathin lamellipodia of migrating fibroblasts have been determined-its thickness in living cells (176 +/- 14 nm), by standing-wave fluorescence microscopy, and its F-actin density (1580 +/- 613 microm of F-actin/microm(3)), via image-based photometry. In combination with data from previous studies, we have computed the density of growing actin filament ends at the lamellipodium margin (241 +/- 100/microm) and the maximum force (1.86 +/- 0.83 nN/microm) and pressure (10.5 +/- 4.8 kPa) obtainable via actin assembly. We have used cell deformability measurements (. J. Cell Sci. 44:187-200;. Proc. Natl. Acad. Sci. USA. 79:5327-5331) and an estimate of the force required to stall the polymerization of a single filament (. Proc. Natl. Acad. Sci. USA. 78:5613-5617;. Biophys. J. 65:316-324) to argue that actin assembly alone could drive lamellipodial extension directly. PMID- 10465782 TI - Monte Carlo simulation of the heterotypic aggregation kinetics of platelets and neutrophils. AB - The heterotypic aggregation of cell mixtures or colloidal particles such as proteins occurs in a variety of settings such as thrombosis, immunology, cell separations, and diagnostics. Using the set of population balance equations (PBEs) to predict dynamic aggregate size and composition distributions is not feasible. The stochastic algorithm of Gillespie for chemical reactions (. J. Comput. Phys. 22:403-434) was reformulated to simulate the kinetic behavior of aggregating systems. The resulting Monte Carlo (MC) algorithm permits exact calculation of the decay rates of monomers and the temporally evolving distribution of sizes and compositions of the aggregates. Moreover, it permits calculation of all moments of these distributions. Using this method, we explored the heterotypic aggregation of fully activated platelets and neutrophils in a linear shear flow of shear rate G = 335 s(-1). At plasma concentrations, the half lives of homotypically aggregating platelet and neutrophil singlets were 8.5 and 2.4 s, respectively. However, for heterotypic aggregation, the half-lives for platelets and neutrophils decreased to 2.0 and 0.11 s, respectively, demonstrating that flowing neutrophils accelerate capture of platelets and growth of aggregates. The required number of calculations per time step of the MC algorithm was typically a small fraction of Omega(1/2), where Omega is the initial number of particles in the system, making this the fastest MC method available. The speed of the algorithm makes feasible the deconvolution of kernels for general biological heterotypic aggregation processes. PMID- 10465783 TI - Whose end is destruction: cell division and the anaphase-promoting complex. PMID- 10465784 TI - ECSIT is an evolutionarily conserved intermediate in the Toll/IL-1 signal transduction pathway. AB - Activation of NF-kappaB as a consequence of signaling through the Toll and IL-1 receptors is a major element of innate immune responses. We report the identification and characterization of a novel intermediate in these signaling pathways that bridges TRAF6 to MEKK-1. This adapter protein, which we have named ECSIT (evolutionarily conserved signaling intermediate in Toll pathways), is specific for the Toll/IL-1 pathways and is a regulator of MEKK-1 processing. Expression of wild-type ECSIT accelerates processing of MEKK-1, whereas a dominant-negative fragment of ECSIT blocks MEKK-1 processing and activation of NF kappaB. These results indicate an important role for ECSIT in signaling to NF kappaB and suggest that processing of MEKK-1 is required for its function in the Toll/IL-1 pathway. PMID- 10465785 TI - Indian hedgehog signaling regulates proliferation and differentiation of chondrocytes and is essential for bone formation. AB - The mechanisms that control cell proliferation and cell differentiation during morphogenesis of the endochondral skeleton of vertebrates are poorly understood. Indian hedgehog (Ihh) signaling from prehypertrophic chondrocytes has been implicated in the control of chondrocyte maturation by way of feedback control of a second secreted factor parathyroid hormone-related peptide (PTHrP) at the articular surfaces. Analysis of an Ihh null mutant suggests a more extensive role for Ihh in skeletal development. Mutants display markedly reduced chondrocyte proliferation, maturation of chondrocytes at inappropriate position, and a failure of osteoblast development in endochondral bones. Together, the results suggest a model in which Ihh coordinates diverse aspects of skeletal morphogenesis through PTHrP-dependent and independent processes. PMID- 10465786 TI - The cytoplasmic Purkinje onconeural antigen cdr2 down-regulates c-Myc function: implications for neuronal and tumor cell survival. AB - Paraneoplastic cerebellar degeneration (PCD) is a disorder in which breast or ovarian tumors express an onconeural antigen termed cdr2, which normally is expressed in cerebellar Purkinje neurons. This leads to an immune response to cdr2 that is associated with tumor immunity and autoimmune cerebellar degeneration. We have found that cdr2, a cytoplasmic protein harboring a helix leucine zipper (HLZ) motif, interacts specifically with the HLZ motif of c-Myc. Both proteins colocalize in the cytoplasm of adult cerebellar Purkinje neurons, and coimmunoprecipitate from tumor cell lines and cerebellar extracts. cdr2 down regulates c-Myc-dependent transcription in cotransfection assays, and redistributes Myc protein in the cytoplasm. Disease antisera from six of six PCD patients specifically blocked the interaction between cdr2 and c-Myc in vitro. These data indicate that cdr2 normally sequesters c-Myc in the neuronal cytoplasm, thereby down-regulating c-Myc activity, and suggest a mechanism whereby inhibition of cdr2 function by autoantibodies in PCD may contribute to Purkinje neuronal death. PMID- 10465787 TI - The Zw5 protein, a component of the scs chromatin domain boundary, is able to block enhancer-promoter interaction. AB - The scs and scs' elements were proposed to function as chromatin domain boundaries for the 87A7 heat shock locus in Drosophila melanogaster. Here we report the identification and characterization of SBP (scs binding protein), a component of the scs nucleoprotein complex. SBP binds specifically to a 24-bp region of scs in vitro and is associated with scs in vivo. Multiple copies of an oligonucleotide containing the SBP recognition sequence are capable of blocking enhancer-promoter interactions in transgene assays. Mutations in the oligonucleotide that disrupt SBP binding in vitro also eliminate enhancer blocking activity in vivo. We show that SBP is encoded by the zeste-white 5 gene and that mutations in zeste-white 5 reduce the enhancer-blocking activity of the multimerized oligonucleotides. PMID- 10465788 TI - The SPOROCYTELESS gene of Arabidopsis is required for initiation of sporogenesis and encodes a novel nuclear protein. AB - The formation of haploid spores marks the initiation of the gametophytic phase of the life cycle of all vascular plants ranging from ferns to angiosperms. In angiosperms, this process is initiated by the differentiation of a subset of floral cells into sporocytes, which then undergo meiotic divisions to form microspores and megaspores. Currently, there is little information available regarding the genes and proteins that regulate this key step in plant reproduction. We report here the identification of a mutation, SPOROCYTELESS (SPL), which blocks sporocyte formation in Arabidopsis thaliana. Analysis of the SPL mutation suggests that development of the anther walls and the tapetum and microsporocyte formation are tightly coupled, and that nucellar development may be dependent on megasporocyte formation. Molecular cloning of the SPL gene showed that it encodes a novel nuclear protein related to MADS box transcription factors and that it is expressed during microsporogenesis and megasporogenesis. These data suggest that the SPL gene product is a transcriptional regulator of sporocyte development in Arabidopsis. PMID- 10465789 TI - Nuclear export of the small ribosomal subunit requires the ran-GTPase cycle and certain nucleoporins. AB - After their assembly in the nucleolus, ribosomal subunits are exported from the nucleus to the cytoplasm. After export, the 20S rRNA in the small ribosomal subunit is cleaved to yield 18S rRNA and the small 5' ITS1 fragment. The 5' ITS1 RNA is normally degraded by the cytoplasmic Xrn1 exonuclease, but in strains lacking XRN1, the 5' ITS1 fragment accumulates in the cytoplasm. Using the cytoplasmic localization of the 5' ITS1 fragment as an indicator for the export of the small ribosomal subunit, we have identified genes that are required for ribosome export. Ribosome export is dependent on the Ran-GTPase as mutations in Ran or its regulators caused 5' ITS1 to accumulate in the nucleoplasm. Mutations in the genes encoding the nucleoporin Nup82 and in the NES exporter Xpo1/Crm1 also caused the nucleoplasmic accumulation of 5' ITS1. Mutants in a subset of nucleoporins and in the nuclear transport factors Srp1, Kap95, Pse1, Cse1, and Mtr10 accumulate the 5' ITS1 in the nucleolus and affect ribosome assembly. In contrast, we did not detect nuclear accumulation of 5' ITS1 in 28 yeast strains that have mutations in other genes affecting nuclear trafficking. PMID- 10465790 TI - Bacterial promoter architecture: subsite structure of UP elements and interactions with the carboxy-terminal domain of the RNA polymerase alpha subunit. AB - We demonstrate here that the previously described bacterial promoter upstream element (UP element) consists of two distinct subsites, each of which, by itself, can bind the RNA polymerase holoenzyme alpha subunit carboxy-terminal domain (RNAP alphaCTD) and stimulate transcription. Using binding-site-selection experiments, we identify the consensus sequence for each subsite. The selected proximal subsites (positions -46 to -38; consensus 5'-AAAAAARNR-3') stimulate transcription up to 170-fold, and the selected distal subsites (positions -57 to 47; consensus 5'-AWWWWWTTTTT-3') stimulate transcription up to 16-fold. RNAP has subunit composition alpha(2)betabeta'sigma and thus contains two copies of alphaCTD. Experiments with RNAP derivatives containing only one copy of alphaCTD indicate, in contrast to a previous report, that the two alphaCTDs function interchangeably with respect to UP element recognition. Furthermore, function of the consensus proximal subsite requires only one copy of alphaCTD, whereas function of the consensus distal subsite requires both copies of alphaCTD. We propose that each subsite constitutes a binding site for a copy of alphaCTD, and that binding of an alphaCTD to the proximal subsite region (through specific interactions with a consensus proximal subsite or through nonspecific interactions with a nonconsensus proximal subsite) is a prerequisite for binding of the other alphaCTD to the distal subsite. PMID- 10465791 TI - The yeast exosome and human PM-Scl are related complexes of 3' --> 5' exonucleases. AB - We previously identified a complex of 3' --> 5' exoribonucleases, designated the exosome, that is expected to play a major role in diverse RNA processing and degradation pathways. Further biochemical and genetic analyses have revealed six novel components of the complex. Therefore, the complex contains 11 components, 10 of which are predicted to be 3' --> 5' exoribonucleases on the basis of sequence homology. Human homologs were identified for 9 of the 11 yeast exosome components, three of which complement mutations in the respective yeast genes. Two of the newly identified exosome components are homologous to known components of the PM-Scl particle, a multisubunit complex recognized by autoimmune sera of patients suffering from polymyositis-scleroderma overlap syndrome. We demonstrate that the homolog of the Rrp4p exosome subunit is also a component of the PM-Scl complex, thereby providing compelling evidence that the yeast exosome and human PM-Scl complexes are functionally equivalent. The two complexes are similar in size, and biochemical fractionation and indirect immunofluorescence experiments show that, in both yeast and humans, nuclear and cytoplasmic forms of the complex exist that differ only by the presence of the Rrp6p/PM-Scl100 subunit exclusively in the nuclear complex. PMID- 10465794 TI - The demography of centenarians in England and Wales. AB - Using a new database, the article investigates the causes of the current 'explosion' in the numbers of centenarians. It then examines the latest official projections for the future and their implications for the highest ages which are likely to be attained. PMID- 10465792 TI - A role for the Cdc7 kinase regulatory subunit Dbf4p in the formation of initiation-competent origins of replication. AB - Using a reconstituted DNA replication assay from yeast, we demonstrate that two kinase complexes are essential for the promotion of replication in vitro. An active Clb/Cdc28 kinase complex, or its vertebrate equivalent, is required in trans to stimulate initiation in G(1)-phase nuclei, whereas the Dbf4/Cdc7 kinase complex must be provided by the template nuclei themselves. The regulatory subunit of Cdc7p, Dbf4p, accumulates during late G(1) phase, becomes chromatin associated prior to Clb/Cdc28 activation, and assumes a punctate pattern of localization that is similar to, and dependent on, the origin recognition complex (ORC). The association of Dbf4p with a detergent-insoluble chromatin fraction in G(1)-phase nuclei requires ORC but not Cdc6p or Clb/Cdc28 kinase activity, and correlates with competence for initiation. We propose a model in which Dbf4p targets Cdc7p to the prereplication complex prior to the G(1)/S transition, by a pathway parallel to, but independent of, the Cdc6p-dependent recruitment of MCMs. PMID- 10465795 TI - Cohabitational and marital histories of adults in Great Britain. AB - This article presents findings on cohabitation-derived from cohabitation and marriage histories collected in a specially designed module of the ONS Omnibus Survey. It examines the sequence of types of partnerships, and how this sequence varies by birth cohort of respondents. Also compared is the relative stability of cohabiting unions and married partnerships. Finally, the reasons for converting a cohabiting union into a marriage are analysed--separately for men and women, and separately according to whether the marriage continued or ended. PMID- 10465793 TI - A novel p34(cdc2)-binding and activating protein that is necessary and sufficient to trigger G(2)/M progression in Xenopus oocytes. AB - The activation of maturation-promoting factor (MPF) is required for G(2)/M progression in eukaryotic cells. Xenopus oocytes are arrested in G(2) and are induced to enter M phase of meiosis by progesterone stimulation. This process is known as meiotic maturation and requires the translation of specific maternal mRNAs stored in the oocytes. We have used an expression cloning strategy to functionally identify proteins involved in G(2)/M progression in Xenopus oocytes. Here we report the cloning of two novel cDNAs that when expressed in oocytes induce meiotic maturation efficiently. The two cDNAs encode proteins of 33 kD that are 88% identical and have no significant homologies to other sequences in databases. These proteins, which we refer to as p33(ringo) (rapid inducer of G(2)/M progression in oocytes), induce very rapid MPF activation in cycloheximide treated oocytes. Conversely, ablation of endogenous p33(ringo) mRNAs using antisense oligonucleotides inhibits progesterone-induced maturation, suggesting that synthesis of p33(ringo) is required for this process. We also show that p33(ringo) binds to and activates the kinase activity of p34(cdc2) but does not associate with p34(cdc2)/cyclin B complexes. Our results identify a novel p34(cdc2) binding and activating protein that regulates the G(2)/M transition during oocyte maturation. PMID- 10465796 TI - Cohabitation in Western Europe. AB - This article examines for a range of European countries: the extent to which men and women are not forming partnerships and how this has changed over time; the number of partnerships experienced; type of first partnership in terms of whether it commences with marriage or cohabitation and, if the latter, whether it converts into marriage; as well as the duration of different types of cohabiting unions. It also includes an examination of the risk of breakdown amongst first unions that commence with cohabitation or marriage and the variations in type of first union according to background characteristics, including: educational attainment, religious observance and experience of parental divorce in childhood. PMID- 10465797 TI - The ethnic minority populations of Great Britain--latest estimates. AB - This article looks at the latest estimates of the ethnic minority populations living in private households in Great Britain. It also highlights differences between the various ethnic groups in relation to geographical distribution, sex and age structure, people living alone and people born in the UK. PMID- 10465798 TI - Can patient registers give an improved measure of internal migration in England and Wales? AB - This article describes a new source of internal migration data for England and Wales. These data are extracted from the 98 Family Health Service Authority (FHSA) patient registers that hold the details of people in England and Wales who are registered with the National Health Service. Migration estimates are derived from changes in postcodes on the patients' records. The findings presented in the article show that the registers provide high quality estimates of migration into and out of the constituent areas of the FHSAs (as compared to other sources). Where limitations are identified, compensatory adjustments can be introduced to further enhance the quality of the estimates. PMID- 10465799 TI - Statistical methods for epidemiologic studies of the health effects of air pollution. AB - We describe two statistical designs that can provide efficient estimates of the health effects of exposure to air pollutants in epidemiologic studies. We also evaluate the effects of measurement error in exposure assessment on the accuracy of estimated health effects. The bidirectional case-crossover design is a variant of a method proposed by Maclure (1991). Our version of the method takes advantage of the fact that in epidemiologic studies involving environmental exposure, accurate information about past exposure is more readily available, and that levels of exposure are generally unaffected by the response of the subject. It differs from other case-crossover methods in that control information is assessed both before and after failure, thus avoiding confounding due to time trends in exposure. The multilevel analytic design provides a method of combining estimates of health effects made on the individual level with those made at the group level. It has great potential value in situations where variations in exposure within groups may not be great enough to provide adequate power to detect health effects, as is often the case in air pollution studies where exposure levels are similar within a geographic community. Measurement errors in exposure assessment can have substantial impact on the accuracy of estimated health effects. When the microenvironmental approach is used to estimate exposure, a standard error of 30% in estimating indoor/outdoor ratios can increase the standard error of a relative risk estimate by 50%, and introduce bias as well. Similar results hold when exposure is estimated with personal samplers. When the microenvironmental approach is used, errors in estimating indoor/outdoor ratios have more influence on the accuracy of risk estimation than do errors in estimating the time spent in microenvironments. PMID- 10465800 TI - [Arousal level and blink activity]. AB - The purpose of this study was to investigate the relationship between arousal level and blink activity; blink rate, blink amplitude and blink duration. Blink activity was examined under two different tasks in which arousal level was manipulated. One was a vigilance task (Experiment 1) for comparatively high arousal level. The other was a simple counting task in a dark and quiet room (Experiment 2) for extremely lower arousal level. Results indicated that blink rates and blink durations tended to increase as arousal level got lower in the monotonous vigilance task situations, meanwhile, blink rates and blink amplitudes decreased as a function of negative arousal level in the pre-sleep situation. PMID- 10465801 TI - [Spontaneous formation of generalized exchange system: an experimental study of discriminating altruism]. AB - Recent computer simulation studies (Takahashi & Yamagishi, 1995, 1996) suggest the possibility that altruists outperform egoists, and the key to their advantage is a strategy called Downward OFT. The strategy calls for acting always altruistically, at the same time selecting as the target of the altruism the most altruistic person among potential recipients. The main purpose of this study was to examine whether people actually follow Downward OFT in a situation where no direct reciprocity was possible. Results were as follows: (1) Pure generalized exchange (i.e., unilateral recourse giving) did emerge and persisted over forty five trials. (2) Subjects selected other altruists as recipients of their unilateral resource giving. (3) The tendency of altruistic subjects to discriminate among potential recipients of their altruism was stronger than that of self-interested subjects. (4) Subjects who unilaterally gave more resource ended up with more profit (resource received minus resource given) than those who gave less. PMID- 10465802 TI - [The effects of marital communication and social activity on alienation in married women and men]. AB - Effects of marital communication and friendship- and job-related social activities on emotional support and alienation among married women and men were examined. Causal analyses of questionnaire responses by 259 women and 185 men indicated that for both women and men, richness of marital communication made the spouse more important as the agent of emotional support, and at the same time decreased alienation. Job-related activities decreased women's alienation, while they tended to increase men's alienation. Men's social activities had a negative influence on their marital relation. In short, women and men appeared to seek different psychological functions in social activities, which in turn determined their levels of alienation. PMID- 10465803 TI - [Rats' behavior in the course of aversive conditioning measured by a stabilimeter]. AB - Aversive behaviors of rats to the conditioned stimulus in the course of classical conditioning of a light (CS) and electric-shock (US) were analyzed using a stabilimeter. For the Classical Conditioning (CC) group (n = 5), CS was preceded 5 s to US (l s) using delayed conditioning procedure in the 3 daily sessions each of which was consisted of 30 reinforcing trials. For the US group (n = 5), US (l s) was presented at the same time as CC group without any CS. The results showed that the activity of the CC group during CS increased as the trials progressed, but it was not changed in the US group. Thus, animals made various kinds of activities during the CS, and these were not reflexes to the presentation of the CS but were responses that have long latencies. PMID- 10465804 TI - [Situations and cognitive appraisals in 'agari' experiences: feature analyses of 'agari' experiences]. AB - The purpose of this study was to examine the 'agari' experiences in everyday life. 'Agari' is a Japanese noun (the verb form is 'agaru'), referring to broad experiences including stage-fright, choking under pressure, social anxiety, and so on. Based upon the self-reports of 452 subjects, we constructed, in Study 1, a 52-items Features of 'Agari' Experience Questionnaire (FAEQ). In Study 2, another sample of 364 completed the FAEQ, and factor analysis was performed. The analysis found six primary factors: Self insufficiency, physical insufficiency, trembling, pressure, physiological response, and awareness of others. A second-order factor analysis was performed on them, and two factors emerged: Self-reflection and awareness of self-importance. It may be therefore concluded that looking at the self leads to an 'agari' experience. In addition, hierarchical cluster analysis divided twelve 'agari' situations into four clusters. The clusters were differentiated by two factors of FAEQ. It is suggested that eliciting situations and associated cognitive appraisals differentiated 'agari' experiences. PMID- 10465805 TI - [The evolution of ethical standards in the practice of psychology: a reflection on the APA Code of Ethics]. AB - After briefly describing the need for ethics in the development of professional regulation and analyzing the historical emergence of codes of ethics, the goal of this paper is to scrutinize the process by which the American Psychological Association developed its own Code of Ethics and proceeded to revise it periodically. Different lessons can be derived from these efforts and from the criticisms that were formulated. The need for international standards in professional and research ethics is then considered, and the results of a recent study on this subject are presented. Five major conclusions can be derived from the preceding analysis: (1) Codes of ethics can help professional recognition by stressing the importance given to the protection of the public, (2) the development of a code of ethics is usually related to the advancement of professional practice, (3) ethical standards should be in tune with the cultural values and the belief system of a given community, (4) a well-balanced code should incorporate both general aspirational principles and enforceable standards, and (5) the method used to define principles and standards should be empirically based. PMID- 10465806 TI - Tobacco Control: reflections on the first seven years. PMID- 10465807 TI - Into the next millenium at the "one stop shopping guide" for tobacco control. PMID- 10465808 TI - Where there's smoke, there's fire. PMID- 10465809 TI - The pied pipers of puffing. PMID- 10465810 TI - Senegal: birth of a new tobacco control group. PMID- 10465811 TI - Uganda: BAT soccer deal goes down to health. PMID- 10465812 TI - Global assessment of deforestation related to tobacco farming. AB - OBJECTIVES: To assess the global amount of forest and woodland consumed annually for curing tobacco between 1990 and 1995; to estimate tobacco's share in total deforestation; to rank tobacco-growing countries by the degree of impact of tobacco deforestation; and to indicate environmental criticality emerging from tobacco's impact on forest resources. DESIGN: Production of country-specific estimates of forests/woodlands needed and depleted on the basis of growing stock/increment of woody biomass involved and wood consumption of tobacco. Comparison of results with secondary statistics on forest cover, deforestation, and population development. RESULTS: An estimated 200,000 ha of forests/woodlands are removed by tobacco farming each year. Deforestation mainly occurs in the developing world, amounting to 1.7% of global net losses of forest cover or 4.6% of total national deforestation. Environmental criticality exists or is emerging in 35 countries with an estimated serious, high, and medium degree of tobacco related deforestation, mainly in southern Africa, middle east, south, and east Asia, South America, and the Caribbean. CONCLUSION: The hypothesis that deforestation from tobacco production does not have a significant negative effect has to be challenged. For empirical validation, the globally significant pattern of estimated tobacco-related environmental damage ought to be included in international research agendas on global environmental change, to become an integral and rational part of tobacco control policy. PMID- 10465813 TI - Features of sales promotion in cigarette magazine advertisements, 1980-1993: an analysis of youth exposure in the United States. AB - OBJECTIVE: To examine the presence of features of sales promotion in cigarette advertising in United States magazines, and to describe trends in youth (ages 12 17) exposure to such advertising (termed "promotional advertising"). DESIGN: Analysis of 1980-1993 annual data on: (a) total pages and expenditures for "promotional advertising" (advertising that contains features of sales promotion) in 36 popular magazines (all magazines for which data were available), by cigarette brand; and (b) readership characteristics for each magazine. We defined promotional advertising as advertisements that go beyond the simple advertising of the product and its features to include one or more features of sales promotion, such as coupons, "retail value added" promotions, contests, sweepstakes, catalogues, specialty item distribution, and sponsorship of public entertainment or sporting events. MAIN OUTCOME MEASURES: Total pages of, and expenditures for promotional advertising in magazines; and gross impressions (number of readers multiplied by the number of pages of promotional advertising) among youth and total readership. RESULTS: During the period 1980-1993, tobacco companies spent $90.2 million on promotional advertising in the 36 magazines. The proportion of promotional advertising appearing in "youth" magazines (defined as magazines with a greater than average proportion of youth readers) increased from 7% in 1980 to nearly 100% in 1987. Although youth readers represented only 19% of magazine readers, the proportion of youth gross impressions to total gross impressions of tobacco promotional advertising exceeded this value throughout the entire period 1985-1993, peaking at 33% in 1987. The five "youth" cigarette brands (defined as brands smoked by at least 2.5% of smokers aged 10-15 years in 1993) accounted for 59% of promotional advertising in all magazines, but for 83% of promotional advertising in youth magazines during the study period. CONCLUSIONS: In their magazine advertising, cigarette companies are preferentially exposing young people to advertisements that contain sales promotional features. PMID- 10465814 TI - Sharing the blame: smoking experimentation and future smoking-attributable mortality due to Joe Camel and Marlboro advertising and promotions. AB - BACKGROUND: Despite public denials, internal tobacco company documents indicate that adolescents have long been the target of cigarette advertising and promotional activities. Recent longitudinal evidence suggests that 34% of new experimentation occurs because of advertising and promotions. OBJECTIVE: To apportion responsibility for smoking experimentation and future smoking attributable mortality among major cigarette brands attractive to young people (Camel and Marlboro). DATA SOURCES, SETTING, AND PARTICIPANTS: Data were from confirmed never-smoking adolescents (12-17 years old) responding to the 1993 (n = 2659) and 1996 (n = 2779) population-based California Tobacco Surveys. MAIN OUTCOMES: Adolescents named the brand of their favourite cigarette advertisements and tobacco promotional items. Using these "market shares" and the relative importance of advertising and promotions in encouraging smoking, we estimated how many new experimenters from 1988 to 1998 in the United States can be attributed to Camel and Marlboro. From other data on the natural history of smoking, we projected how many future deaths in the United States can be attributed to each brand. RESULTS: Although Camel advertisements were favoured more than Marlboro and other brands in 1993 and 1996, the "market share" for promotional items shifted markedly during this period from Camel and other brands towards Marlboro. We estimated that between 1988 and 1998, there will be 7.9 million new experimenters because of tobacco advertising and promotions. This will result in 4.7 million new established smokers: 2.1, 1.2, and 1.4 million due to Camel, Marlboro, and other brands' advertising and promotions, respectively. Of these, 1.2 million will eventually die from smoking-attributable diseases: 520,000 from Camel, 300,000 from Marlboro, and the remainder from other brands. CONCLUSIONS: Our analysis provides a reasonable first estimate at sharing the blame for the long-term health consequences of smoking among the major brands that encourage adolescents to start smoking. PMID- 10465816 TI - Prevalence and determinants of smoking in three regions of Saudi Arabia. AB - OBJECTIVES: To study the prevalence and determinants of cigarette smoking among Saudi nationals in three regions of Saudi Arabia. PARTICIPANTS: A sample of 8310 individuals aged 15 years and above from both sexes, randomly selected from the three regions, using a stratified cluster sampling technique. DESIGN: A cross sectional, household, community-based survey. Using a predesigned and tested questionnaire, the participants were interviewed by primary care physicians. The interview covered personal, social, and educational characteristics of the respondents, and also included questions about their smoking status, duration of smoking, and daily cigarette consumption. MAIN OUTCOME MEASURES: Association between current smoking and sociodemographic variables, in univariate and multivariate analysis. Degree of interaction between the different determinants of cigarette smoking. RESULTS: The overall prevalence of current smoking was 21.1% for males and 0.9% for females. Most smokers (78%) were young to middle aged (21-50 years old). Smoking prevalence was higher among married people, among uneducated people, and among those in certain occupations: manual workers, businessmen, army officers, and office workers. CONCLUSIONS: Cigarette smoking is an important public health problem in Saudi Arabia. A more intense and comprehensive tobacco control effort is needed. PMID- 10465815 TI - Psychosocial predictors of cigarette smoking among adolescents living in public housing developments. AB - BACKGROUND: Adolescents residing in low-income public housing developments in inner-city regions may be particularly vulnerable to a variety of risk factors associated with cigarette smoking. OBJECTIVE: To elucidate the aetiology of cigarette smoking among adolescents living in public housing developments. DESIGN, SETTING, AND SUBJECTS: We examined predictors of smoking from four domains: background characteristics, social influences, behavioural control, and psychosocial characteristics using a sample of seventh graders (mean age 12.9 years) who reside in public housing developments in New York City (n = 624). The addresses of participants in a larger investigation of the aetiology and prevention of smoking were checked to determine if they lived in one of 335 public housing developments in New York City. All participants living in public housing developments were included in the current study. MAIN OUTCOME MEASURES: African-American and Hispanic students completed questionnaires about their cigarette use, social pressures to smoke, smoking attitudes, smoking knowledge, and smoking resistance skills. Students also provided information on demographic and behavioural control (such as church and school attendance). RESULTS: Logistic regression analyses indicated that social influences from friends and family members predicted smoking. Psychosocial characteristics such as advertising resistance skills, anti-smoking attitudes, and refusal skills lowered the odds of smoking. CONCLUSIONS: These findings suggest that smoking prevention approaches targeted at these young people should increase their awareness of social pressures to smoke, correct misperceptions about the prevalence of smoking among friends, and teach relevant psychosocial skills. PMID- 10465817 TI - Comparison of two self-help smoking cessation booklets. AB - OBJECTIVE: To compare two self-help smoking cessation booklets distributed to callers to a Quitline telephone service in Queensland (Australia). DESIGN: Callers were randomised to receive either a structured 14-day quit programme (Time to quit) or another booklet and described four broad stages of quitting (Can quit). Approximately one month later, these callers were interviewed by telephone. MAIN OUTCOME MEASURES: Self-reported smoking status at one month and recent quit attempts together with process measures. RESULTS: Altogether, 521 callers (78.3%) were interviewed. They were heavier smokers when compared with all Queensland smokers: on average they had smoked for more than 15 years, smoked nearly 25 cigarettes per day, and almost two-thirds had attempted to quit smoking in the past year. In each group, significant proportions either did not begin to use the booklet (50.5-56.0%), or did not complete its use (77.4-82.3%). There were no differences in the self-reported quit rates at one month (17.0% vs 16.1%; p = 0.93). In an ordinal regression modelling procedure involving age, sex, number of recent quit attempts, number of cigarettes smoked per day, smoking status of partner, number of five closest friends who smoke, education, and booklet received, only the number of cigarettes smoked per day was significantly related to smoking status at one month. CONCLUSIONS: Callers to telephone Quit line services are typically heavier smokers than the general smoking population, and simple strategies, such as self-help booklets, appear to achieve relatively high success. Nevertheless, there is potential to improve the effectiveness of these materials by making a range of materials available and encouraging callers to make a serious attempt to quit smoking. PMID- 10465818 TI - Attitudes and experiences of restaurateurs regarding smoking bans in Adelaide, South Australia. AB - OBJECTIVES: To determine compliance with a voluntary code of practice (VCP) for restricting smoking in restaurants and to canvass the attitudes of restaurateurs towards tougher smoking restrictions. DESIGN: Cross-sectional survey conducted in 1996 using a telephone questionnaire. SETTING: Metropolitan restaurants and cafes in Adelaide, South Australia. PARTICIPANTS: 276 (86.8%) of a sample of randomly selected owners and managers. MAIN OUTCOME MEASURES: Restaurant non-smoking policies, reported and anticipated change in business, and restaurateurs' attitudes towards smoking restrictions. RESULTS: 26.8% of restaurants had a total smoking ban; 40.6% restricted smoking some other way; and 32.6% permitted unrestricted smoking. Only 15.1% of restaurants with a ban or restrictions had used the VCP to guide the development of their policy, and only half of these were complying with it. Although 78.4% of those with bans and 84.4% of those with restrictions reported that their non-smoking policy had been associated with either no change or a gain in business, only 33.3% of those allowing unrestricted smoking expected that this would be the case, if they were to limit smoking. A total of 50.4% of restaurateurs, including 45.3% of those with no restrictions, agreed that the government should ban smoking in all restaurants. CONCLUSIONS: The VCP made an insignificant contribution to adoption of non-smoking policies, and compliance with the code was poor. Despite concerns about loss of business, there was considerable support for legislation which would ban smoking in all dining establishments. PMID- 10465819 TI - How the tobacco industry continues to keep the home fires burning. PMID- 10465820 TI - Institutional addiction to tobacco. PMID- 10465821 TI - Consumption and production waste: another externality of tobacco use. AB - OBJECTIVE: To describe the waste produced by and environmental implications of individual cigarette consumption (filter tips, packages, and cartons) and tobacco manufacturing. STUDY SELECTION: All available articles and reports published since 1970 related to cigarette consumption and production waste were reviewed. DATA SOURCES: Global cigarette consumption data were used to estimate cigarette butt and packaging waste quantities. Data from the Center for Marine Conservation's International Coastal Cleanup Project were used to describe some environmental impacts of tobacco-related trash. Data from the United States Environmental Protection Agency's (EPA's) Toxics Release Inventory and reported global cigarette consumption totals were used to estimate waste production from cigarette manufacturing. DATA EXTRACTION AND SYNTHESIS: In 1995, an estimated 5.535 trillion cigarettes (27,675 million cartons and 276,753 million packages) were sold by the tobacco industry globally. Some of the wastes from these products were properly deposited, but a large amount of tobacco consumption waste ends up in the environment. Some is recovered during environmental clean-up days. For the past eight years (1990-1997), cigarette butts have been the leading item found during the International Coastal Cleanup Project; they accounted for 19.1% of all items collected in 1997. The tobacco manufacturing process produces liquid, solid, and airborne waste. Among those wastes, some materials, including nicotine, are designated by the EPA as Toxics Release Inventory (TRI) chemicals. These are possible environmental health hazards. In 1995, the global tobacco industry produced an estimated 2262 million kilograms of manufacturing waste and 209 million kilograms of chemical waste. In addition, total nicotine waste produced in the manufacture of reduced nicotine cigarettes was estimated at 300 million kilograms. CONCLUSIONS: Laws against littering relative to cigarette butts could be better enforced. Additional taxes might be levied on cigarette products that would then be directed to environmental clean-up efforts. The tobacco industry should improve the biodegradability of filters, reduce packaging waste, and educate its customers. Worksites and public buildings should be encouraged or required to supply appropriate disposal mechanisms at all building entrances. Public awareness campaigns about the magnitude and prevention of cigarette consumption waste could be developed through partnerships among environmental groups, health organisations, and environmental protection agencies. Tobacco production waste should be a source of concern and regulation by governments throughout the world; it contains numerous chemicals which may be considered health hazards, not the least of which is nicotine produced in the manufacture of low-nicotine cigarettes. PMID- 10465822 TI - The 1998 Subic Bay international declaration of support for action to control tobacco. Asia Pacific Association for the Control of Tobacco (APACT). PMID- 10465823 TI - Tobacco sponsorship is no laughing matter. PMID- 10465824 TI - Tobacco advertising in Indonesia: "the defining characteristics for success". PMID- 10465825 TI - The fourth largest market in the world. PMID- 10465826 TI - Reflections: testifying in the Minnesota tobacco lawsuit. PMID- 10465827 TI - Eliminating nicotine in cigarettes. PMID- 10465828 TI - Eliminating nicotine in cigarettes. PMID- 10465829 TI - Eliminating nicotine in cigarettes. PMID- 10465830 TI - "Schools, Internet, and nonsmoking": the use of new media in tobacco control and health promotion. PMID- 10465831 TI - A new smoking cessation programme using the Internet. PMID- 10465832 TI - Cigarette taxes. PMID- 10465833 TI - Changes in the focus of cigarette advertisements in the 1950s. PMID- 10465835 TI - [The characteristics of osteoclast localization in the bones of the extremities of the crested salamander Pleurodeles waltlii]. AB - Localization of polynuclear osteoclasts in limb bones of ribbed newt were analyzed in consideration of calcium content in different bone structures. With using the methods of light and scanning electron microscopy and X-ray microanalysis it was shown that a critical level of cartilage calcification is need to activate osteoclasts. Osseous tissue can bind more calcium salt without resorption. PMID- 10465837 TI - [The frequency of spontaneous apoptosis in the peripheral blood lymphocytes of tumor carriers as a criterion in the prognosis of the efficacy of special treatment]. AB - The relationship between level of spontaneous apoptosis in the lymphocytes of peripheral blood of tumor patients, detected before the treatment, and effectiveness of following special therapy was found. The possibility of using of the proposed method of spontaneous lymphocytes apoptosis detection for the objective prescribing of chemotherapy was demonstrated. PMID- 10465838 TI - [CD44 gene expression in cancerous thyroid cells]. AB - The peculiarities of alternative CD44 mRNA splicing in thyroid cancer tissue of children from radiocontaminated areas was investigated. CD44 gene expression in thyroid cancer tissues of children exposed to radiation resembled that in spontaneously emerged cancers. It was concluded that CD44 gene expression is not the primary target of radioactive irradiation. Probably, the CD44 mRNA splicing deregulation is the consequence of cancer. PMID- 10465845 TI - [The genetic structure of Kulunda sheep]. AB - The analysis of the genetic structure of the Kulunda sheep in eight polymorphous genetic-biochemical systems was carried out. While comparing with the genetic structures of number of sheep breeds, differentiated by the origin history, productivity traits and the breeding regions, the locus-specificity particularities of the Kulunda sheep were described. The importance of creating of the "genetic portrait" for disappearing breeds was discussed. PMID- 10465846 TI - [The construction and use of inosine-containing primers for searching for and identifying the genes of insecticidal crystal proteins in Bacillus thuringiensis]. AB - The specific to 3 types of Cry genes primers containing inosine were constructed to find crystal insecticidal protein Bacillus thuringiensis genes using PCR. A number of new B. thuringiensis strains isolated in Ukraine were investigated using these PCR primers. As a results, Cry genes were found, some of them were identified and demonstrated high homology to Cry1Ba2 and Cry1Bc genes. PMID- 10465847 TI - [Chromosome association with the nuclear membrane and the orderliness of the spatial organization of the genetic material in the interphase nucleus]. AB - The role of chromosome-membrane associations in order of spatial chromosome organization in interphase nuclear is reviewed. Data on specificity and mechanism of interphase chromosome anchoring on nuclear membrane are summarized. PMID- 10465848 TI - [Steroidogenesis in epithelial cells of rat epididymis]. AB - Studies were performed on cultured epithelial cells of the caput and cauda of epididymis stemming from male rats of inbred Wistar strain. The cultures were conducted on a full medium enriched with 5% fetal calf serum in the presence or without exogenic androgens-T and DHT. The cells were identified by means of immunohistochemical reactions with the use of monoclonal antibodies against cytokeratin and desmin (Fig. 1, 2). All cells in the culture showed positive reaction to cytokeratin. At the same time there was a lack of desmin-positive cells. Through secreting the proteins, glycoproteins, glycolipids, phospholipids and number of other substances the epithelial cells of epididymis create an environment for maturating and storing of spermatozoa in lumen of the duct. Synthesis of these substances is possible thanks to the expression of genes defined for a given zone of epididymis, the expression being mainly regulated by androgen, although a share of estrogens is also evidenced in this process. The cytoplasm of epithelial cells of epididymis fails to reveal the presence of secretory granules, while the mechanism of releasing the secretion still continues to be controversial. There are also some and unverified suggestions about the capability of these cells to synthesize androgens. In connection with what was mentioned above, the objective of the work was to establish the mode of releasing the secretions by cultured epithelial cells of epididymis as well as to determine whether these cells synthesize androgens and if they may be the source of estrogens. Electron-microscopic observations disclosed a rich content of rough endoplasmic reticulum and structures similar to secretory units produced from concentrically arranged membranes encircling cytoplasm fragments in their interior (Fig. 10B, 14, 15A). There were protrusions of cytoplasm on the surface of cells. Released secretion was present between the cells. The apocrine way of releasing was confirmed also by scanning electron microscope. Numerous granular protrusions were released into the intercellular space (Fig. 19). The process of synthesis and release of secretion was androgen-dependent. Cells cultured without addition of exogenic androgens were characterized by disorganization of organelles and reduction of their number, particularly rough endoplasmic reticulum. The surface of cells was prevalently smooth, deprived from protrusions (Fig. 21). Very close neighbourhood, and sometimes a direct contact of lipid droplets and mitochondria with lamellar cristae as well as the presence of smooth endoplasmic reticulum observed in cytoplasm of cultured epithelial cells of epididymis, suggest their similarity to steroidogenic cells (Fig. 11A, 12). This is also indicated by the finding that these cells reveal the presence of active enzymes of the steroidogenesis pathway, 3 beta-HSD and 17 beta-HSD exhibited in histochemical reactions (Fig. 8, 9). RIA determination of hormones in the medium, wherein the epithelial cells had been cultured showed that the said cells synthesized and released DHEA, A and T, but in low and sometimes trace concentrations (Tab. 1-3). Lack of progesterone in medium of the cells on the 3rd and 5th days of culture indicates that the synthesis of testosterone and earlier forms of androgens proceeds using delta 5 metabolites, as it takes place in human testis. The cells' medium on the 3rd and 5th days of culture was found to disclose high concentration of 17 beta-estradiol (E2) (Tab. 4). E2 concentrations were always higher when the cells were grown without the addition of exogenic androgens. In this cases the cytoplasm of the cells displayed depolymerization of microtubules, which enhances the approximation to each other of structures participating in steroidogenesis and translocation of substrates and products of the consecutive stages of steroidogenesis. (ABSTRACT TRUNCATED) PMID- 10465850 TI - Serum iron in young athletes: preliminary results. PMID- 10465849 TI - [Identification of enteroviruses in environmental samples using PCR coupled with a colorimetric microwell method]. PMID- 10465851 TI - [Prevention of allergic reactions in latex-sensitized patients]. PMID- 10465852 TI - A methodological approach for a nutritional surveillance system in a transition country: the case of Armenia. PMID- 10465853 TI - [Study on smoking of graduate students at the School of Medicine and Surgery of the Chieti University: analysis of associations with socio-environmental variables]. PMID- 10465854 TI - [Smoking habits and opinions of a sample of high school students]. PMID- 10465855 TI - [Youth law violations in the city of Cagliari: an analysis of risk in small urban areas]. PMID- 10465856 TI - [Application of the C.C.N.L, in the management if National Health Service. Experience of the USL 2 Pentria Agency in the introduction of the MBO (Management by Objectives) System]. PMID- 10465857 TI - [Health organizations: how to attract and keep the best minds?]. PMID- 10465858 TI - Drug development for women. PMID- 10465859 TI - Therapeutic equivalence: all studies are not created equal. AB - With an increasing number of available treatment options, clinicians must frequently evaluate whether comparable therapies are equivalent in terms of efficacy and safety. Two methodologically distinct study designs are used to establish therapeutic equivalence: standard superiority trials and true equivalence trials. In either study design, clinician-readers assess equivalence by examining both the statistical significance and the clinical importance of the study results (as defined by the minimally important difference, the smallest difference in patient outcome that would lead to an important difference in patient health status). Once therapeutic equivalence has been established, clinicians may select one therapy as the preferred treatment option because it offers other clinical benefits, such as a lower cost or a more convenient drug administration schedule. PMID- 10465860 TI - Hypoglycemia associated with high doses of sertraline and sulphonylurea compound in a noninsulin-dependent diabetes mellitus patient. AB - Unlike other selective serotonin reuptake inhibitors (SSRIs), sertraline has linear pharmacokinetics so that increases in dose lead to proportional increases in drug concentration. The half-life of sertraline is about 26 h so that it reaches a steady state in one week, according to the product monograph. Hypoglycemia associated with sertraline and coadministration of oral hypoglycemics belonging to the sulphonylurea derivatives has rarely been reported. A patient with schizoaffective disorder with non-insulin-dependent diabetes mellitus (NIDDM) treated with sertraline, risperidone and glyburide who developed hypoglycemia is presented. The article highlights that inhibition of P450 enzymes can be affected by several different factors. Interactions are possible whenever a patient concomitantly receives two drugs that bind to the same P450 system Greater inhibition was likely induced at doses higher than those recommended. This process was reversed within 10 days of discontinuing the sertraline. Good glycemic control followed discontinuation of psychotropic drugs and the oral hypoglycemic agent. Knowledge of the individual P450 enzymes is important in the metabolism of individual drugs, together with an understanding of the patient's drug metabolizing ability. These factors may lead to more appropriate prescribing and further research into specific P450 enzymes responsible for metabolism of particular drugs, which remains unclear. PMID- 10465861 TI - A possible tetracycline-risperidone-sertraline interaction in an adolescent. AB - A 15-year-old adolescent male, with Asperger's disorder, Tourette's disorder and obsessive-compulsive disorder, on a tetracycline-risperidone-sertraline treatment regimen, had an acute exacerbation of motor and vocal tics. The increase in tics may have resulted from either a tetracycline-risperidone interaction leading to decreased levels of risperidone, or a tetracycline-sertraline interaction leading to increased levels of sertraline or the natural course of the Tourette's disorder. The sertraline dose was increased with no concomitant increase in tics, and subsequent discontinuation of tetracycline resulted in an improvement in tics, which indicates an interaction between tetracycline and risperidone. The addition of antibiotics to psychotropic medications requires close monitoring due to the potential interactions. PMID- 10465862 TI - Risk of drug dependence and abuse posed by barbiturate-containing analgesics. AB - Concern has been raised that the barbiturate component of barbiturate-containing analgesics constitutes a public and individual health problem because of information from literature before 1966 concerning preclinical and clinical abuse liability, and the dependence risk of barbiturates. The safety of barbiturates alone and in combination in analgesics was reviewed. In addition, information from manufacturers of combination products were evaluated. A meta-analysis was not possible because of the paucity of formal clinical trials. Even though barbiturates have a narrow margin of safety and substantial abuse potential, there is no evidence that barbiturate-containing analgesics without codeine represent a public health or social problem because of their abuse potential. However, proper studies to confirm this theory have not been performed. In the absence of better data concerning efficacy and lack of dependence potential, barbiturate-containing analgesics are not first-line medications for the initiation of treatment for pain. Codeine-containing combination analgesics have the potential to be a more important public health problem than those with only barbiturate in the combination. PMID- 10465863 TI - Comparative efficacy and safety of inhaled corticosteroids in asthma. AB - Current guidelines emphasize the efficacy of inhaled corticosteroids for anti inflammatory activity in asthma, and recommend higher doses and earlier initiation of therapy than previous guidelines. Concern over possible side effects with long term use has prompted an evaluation of the available literature to determine the optimal dose that may be administered without fear that significant side effects might occur (e.g., growth retardation in children, adrenal suppression, reduction in bone mineral density, cataract formation). Regular treatment with the following drugs in adults and children, respectively, is unlikely to result in any clinically significant effects on the above parameters: beclomethasone dipropionate less than 1500 micrograms and 400 micrograms, budesonide less than 1600 micrograms and 400 micrograms, flunisolide less than 2000 micrograms and 1000 micrograms, fluticasone propionate approximately 500 micrograms and 200 micrograms, and triamcinolone acetonide less than 1600 micrograms and 1200 micrograms. Systemic effects are influenced by potency and bioavailability. Inhaled corticosteroids owe their favourable safety profile to a high topical to systemic potency ratio compared with that of oral corticosteroids. In terms of relative topical potency, fluticasone propionate is more potent than budesonide, which is more potent than beclomethasone dipropionate, which is more potent than flunisolide and triamcinolone acetonide. The delivery device has an important influence on the amount of drug reaching the patient. A spacer device attached to a metered dose inhaler or a Turbuhaler reduces oropharyngeal deposition and increases lung deposition. As a result, a dosage reduction may be possible, and local side effects of dysphonia and oral candidiasis may be reduced. Patients requiring continued high doses by the inhaled route should be monitored for systemic effects and be considered for osteoporosis prevention therapy if appropriate. PMID- 10465864 TI - [Experience in a hygienic assessment of problems related to physical environmental factors]. PMID- 10465865 TI - [Use of biological properties of planktonic microorganisms in characterizing sanitary status of fresh-water reservoirs]. PMID- 10465866 TI - [Impact of the quality of drinking water on the morbidity of the Yaroslavl population]. PMID- 10465867 TI - [Dynamics of air and drinking water pollution in the Samara Region]. PMID- 10465868 TI - [Occupational diseases of workers at the Magnitogorsk metallurgy enterprise]. PMID- 10465869 TI - [Evaluation of the effects of noise on the functional state of the body]. PMID- 10465870 TI - [Substantiation of maximum allowable concentration of TZ-85 in workplace air in agriculture]. AB - The plant growth stimulant TZ-85 is a native preparation. Its LD50 are as follows: 780, 5000, and 2500 mg/kg for albino mice, albino rats, and rabbits, respectively. The cumulative coefficient of the agent is more than 3; Limac is 235 mg/kg, Limch is 10 mg/kg. Its MAC for the air of a working area is 3 mg/kg. PMID- 10465871 TI - [Characterization of the content of polycyclic aromatic hydrocarbons in the cold- and hot-smoked fish]. AB - The carcinogenic danger of the commercial smoked herring was assessed. The edible part of the smoked herring contains 16 polycyclic aromatic hydrocarbons whose bulk consists of phenanthrene, pyrene, floranthene, and benzo(b)fluoranthene. The content of benzo(a)pyrene (BP) in the cold and hot smoked herring are 189 and 291 ng/kg, respectively. It is lower than the present standard values. The proportion of carcinogenic danger of BP is 32% for cold smoked herring and 34% for hot smoked herring. PMID- 10465872 TI - [Physiologic and health aspects of first-grade schoolchildren's adaptation to school]. PMID- 10465873 TI - [Hygienic and ecological training of pediatricians]. AB - The hygienic training of pediatricians is very important in terms of introducing research achievements into practical public health service. Knowledge must be permanently replenished in the system of postgraduate training by giving up the job for 4 months every 5 years, including an obligatory monthly training in ecological and hygienic problems. The licensing system of the specialists' right to professional activity requires to be introduced. The qualification grade of a pediatrician and the payment of his/her work should be determined by a set of specific licences. PMID- 10465874 TI - [Health status of children and adolescents and risk factors in a geographic endemic area]. AB - The health status of children and adolescents residing in geographic endemic area of the Republic of Tatarstan was studied. Medical examinations of 5078 children and adolescents were performed. They showed a significant increase in the incidence of thyroid hyperplasia caused by iodine deficiency. The successively significant risk factors are biological, naturally ecological, and social. PMID- 10465875 TI - [Role of technogenic pollution as a risk factor in morbidity of preschool children]. AB - The paper shows it adequate to apply a systems approach to assessing the role of technogenic pollution of the environment in the system of socio-hygienic and biomedical factors that affect the morbidity among pre-school children by using the epidemiological survey in the area of the Revdinsk-Pervouralsk industrial center as an example. PMID- 10465876 TI - [Medical and hygienic aspects of thallium neurotoxicity]. PMID- 10465877 TI - [Effects of a plant preparation Korrda-K on metabolic reactions in the liver of rats exposed to acetone]. AB - The paper deals with the study of the preventive effect of the plant drug Korrda K on lipid and carbohydrate metabolism in the liver of the rat exposed to acetone inhalation. The agent was found to have hepatic and membranous protective effects. On exposure to acetone, Korrda-K promotes the normalization of blood glucose levels and other biochemical parameters. The magnitude of changes in the composition of lipids and fatty acids suggests that the preventive use of the agent favours the preservation of the membrane structure of hepatocytes changed by acetone. PMID- 10465878 TI - [State of energy metabolism and microsomal monooxygenases in animals exposed to inhaled 1,2,4-trimethylbenzene]. AB - The investigations demonstrate that unfavourable changes occur in the tissues and red blood cells of the animals exposed to a single dose of 1,2,4-trimethylbenzene (TMB) in a concentration of 10 mg/m3; moreover, the inclusion of 32P is increased in the biochemical structures of the animals and the concentrations of Na, K, Mg, and Ca ions change. Exposure to TMB, 0.01-1.0 mg/m3 causes profound changes in the animals. Microsomal monooxygenases were found to activate in the lung, liver, and kidney of the animals exposed to TB in concentrations of 10 and 1 mg/m3. PMID- 10465879 TI - [Effects of radon on the development of malignant neoformations of the lungs in residents of the Sverdlovsk region]. PMID- 10465880 TI - [Experience at the State Sanitary and Epidemiological Surveillance Center in the Republic of Dagestan]. PMID- 10465881 TI - [Use of a computerized information-modeling system in the evaluation of health hazards of formaldehyde]. PMID- 10465882 TI - [Methods of risk assessment in transit of chemicals from water to air in disaster situations]. PMID- 10465883 TI - [Separate determination of penicillin and gentamycin in the work area air by thin layer chromatography]. PMID- 10465884 TI - [An alternative approach to the classification of hazards of industrial waste]. PMID- 10465885 TI - [Spectrophotometric and photometric determination of 1,4-phenylenediamine in environmental objects]. PMID- 10465886 TI - [Immunoenzyme method of determination of savinase and caresime in the air]. PMID- 10465887 TI - [Methods of determination of tributyltin in water and soil]. PMID- 10465888 TI - [Determination of formaldehyde in biological fluids]. PMID- 10465889 TI - [Environmental transformation of methyl ether]. PMID- 10465890 TI - [Methods and practice of controlling levels of heavy metals in biological samples]. PMID- 10465891 TI - [Significant and anniversary dates of the history of hygiene and sanitation in 1999]. PMID- 10465892 TI - [Splicing and stability of intron in the expression retroviral vector with human clotting factor IX]. AB - To study the role of intron in the expression of hFIX, retroviral vectors with intron containing hFIX were constructed. It is fundamental for the intron study whether the intron constructed in retroviral vector can be steadily transferred into target cell. First, we constructed two forward-orientation retroviral vectors: G1NaC-i-IX contains the exogenous intron from IL-2, and G1NaC-i'-IX contains the truncated intron I from hFIX gene, covering the splicing donor and acceptor sequences. RT-PCR result indicated that intron in the forward orientation retroviral vector was spliced after packaging in PA317. Then, reverse orientation retroviral vectors G1NaC-i'-IXR and G1NaPAIXi' BAM were constructed, in which the reverse and complimentary sequences of hFIX gene with intron appeared in retroviral RNA. RT-PCR assay combined with ELISA test indicated that intron was retained after packaging and hFIX gene with intron constructed in the reverse-orientation retroviral vector can be transduced intact and expressed hFIX at a high level in vitro. PMID- 10465893 TI - [Studies of meiotic origin of the extra chromosome 21 in Down syndromes detected by using (GT)n polymorphic DNA markers]. AB - The polymorphics of two pericentric (GT)n sequences on the long arm of human chromosome 21 have been analyzed after PCR amplification, PAGE and Ag-staining for the first time in 50 Chinese Han people, and were used to detect meiotic origin of the extra chromosome 21 in Down syndromes. Six and 5 alleles were found in Chinese Han people for D21S215 and D21S120, respectively, with observed heterozygosities of 0.68 and polymorphic information content PIC, 0.67 and 0.65. For 17 Down syndromes whose parental origin of the extra chromosome 21 were known, meiotic origin of the extra chromosome 21 were determined in 16 cases, with 7 and 4 maternal meiosis I and II nondisjunction, 2 and 3 paternal meiosis I and II, respectively. The possible biological significance of the study on origin of the extra chromosome 21 has been discussed. PMID- 10465894 TI - [Studies of foreign gene integeation during embryo early development]. AB - There are still some problems in transgenic animals. Gene transfer, for example, reamins a difficult and costly task for animals, the vectors carrying the gene coding for the proteins of interest are of unpredictable efficiency. Therefore, it is important to identify foreign gene integration in genomose before transferring fertilized eggs to receptors, in order to increase efficiency of producing transgenic animal. In this paper, the construct that mice whey acid protein (WAP) gene promoter directs G-CSF gene was used to microinject fertilized eggs of mice. Fertilized eggs containing foreign gene were measured by using PCR method. The results showed that 100%, 77.7% and 44.4% retentions of foreign gene were achieved in 1, 2 and 8 cell-stage, respectively. Two part homologous recombination fragments were constructed and coinjected in to fertilized eggs of mice. PCR amplification fragment went beyond this homologous recombination area. If foreign gene could not integrate in to genomose, the fragment of PCR amplification could not be produced during embryo development. The results showed that the rationes of foreign gene integrated in to genomoes in 1, 2 and 8 cell stage were 11.1%, 55.5% and 44.4%, respectively. This method might provide us a way to screen transgenic eggs when we use embryo section technique in farm animal. PMID- 10465895 TI - [Targeting of MT-II gene in mouse ES cells]. AB - A MT-II gene targeting vector--pMT-II6.7 with 6.7kb sequences homolog to mMT-II and its flanking region has been transfected into Mespu22 with electroporation. We have got 26 positive clones from 104 G418 and Ganc clones with PCR method. From the karyotype analysis, we found two clones with 84% and 88% normal karyotype. They are clone 5-2 and 8-4. Using Southern analysis, we confirm that the two clones are homologous recombinants. These cell lines still have the pluropotential ability to differentiate both in vitro and in vivo. Now we have got a chimera mouse with cells from clone 8-4. PMID- 10465896 TI - [Origin and differentiation of domestic goose breeds in China, inferred from mitochondrial DNA polymorphism]. AB - Mitochondrial DNAs (mtDNAs) of 138 samples from 11 domestic goose breeds in China were investigated by digesting, with 19 restriction endonucleases. Of 19 enzymes used, seven (Bcl I, Dra I, Eco RV, Hae II, Hinc II, Kpn I, Sac I) detected polymorphic patterns. By combining 27 restriction morphs, 138 individuals were classified into 6 mtDNA haplotypes. Phylogenetic tree was constructed by using UPGMA. There was no shared haplotype between Yili breed and the other 10 breeds. Genetic distance and UPGMA tree also suggested that Yili breed and other breeds came from different ancestors. Yili breed originated from Anser anser and other 10 breeds originated from Anser cygnoides. Restriction morphs digested with 4 enzymes (EcoRV, Hae II, Hinc II and Kpn I) could be used as maternal genetic markers to distinguish the two types of domestic geese. Mitochondrial DNA polymorphism was observed in the ten breeds of Anser cygnoides. Nucleotide diversity (pi), genetic distance between the two types and the average genetic distance among the ten breeds were estimated to be 0.025%, 0.266%, 0.029%, respectively. The breeds with white plume were affected by founder effect when they were formed. Swan domestic geese, Anser cygnoides domesticus, in China might come from two different populations of Anser cygnoides at two different places. PMID- 10465897 TI - [Genetic mapping of the rice telomeric regions through PCR]. AB - By using RAPD primer mediated asymmetric PCR (RM-PCR) method, a new type of molecular marker based on the telomeric repeats sequence was developed. PCR-based genetic mapping of telomeric repeat associated sequences (TASs) was conducted with a rice doubled haploid (DH) population derived from the inter-subspecific cross between indica variety (Zhaiyeqing8) and japonica variety (Jingxil7). Twenty-three loci were mapped onto the genetic map. Of these loci, 12 loci were mapped to the most distal position of eight chromosome arms and some of which may be located in subtelomeric region, five loci were mapped to the approximate positions of centromeric regions and six loci were mapped to other interstitial chromosomal regions. PMID- 10465898 TI - [Obtaining transgenic rice plants and their progenies using Agrobacterium tumefaciens]. AB - Rice (Oriza sativa L.) suspension cells of Taipei 309 were co-cultivated with A. tumefaciens stran EHA101 harbouring binary vector pBYT2 for 3 days in the presence of vir inducer, 100 mumol/L acetosyringone (AS). After 2 months of continuous selection, 17 stable hygromycin-resistant, GUS-positive calli were recovered from 364 suspension cell clusters co-cultivated with A. tumefaciens. 10 putative transgenic R0 plants obtained from 8 tansformed calli and their progenies were analyzed for the integration and expression of foreign genes. Southern blot analysis of R0 and R1 generations indicated that foreign genes had been stably integrated in the genome of transgenic rice and sexually transmitted. One of the transgenic lines showed 5 copies of T-DNA integration, while the others had only one copy. Histochemical staining observation and fluorometric assay of GUS activity in transgenic rice cells and plants showed ubiquitin promoter from maize was highly effective in driving the expression of gus reporter gene in transgenic rice cells. GUS protein and its activity were also investigated through ndPAGE-X-Gluc staining assay, and it was found that the GUS protein in transgenic rice cells was smaller in size than the standard GUS protein (Sigma Co. G0786) but as large as that from E.coli HB101 (pBI121). This study suggested that Agrobacterium-mediated transformation of plant is an efficient and reliable method to introduce foreign genes into rice. PMID- 10465899 TI - [A chromosomal integration system for development of polyvalent vaccine strains]. AB - We have developed a potential bivalent vaccine stain through the chromosomal integration system in which the gene that codes for the B subunit of cholera toxin was integrated into the chromosome of an attenuated strain of Salmonella typhimurium. This system involves two steps: construction of a hisOG deletion mutation into the chromosome of S. typhimurium strain SL3261: and replacement of the hisOG deletion by the complete hisOG region and the segment of heterologous DNA which codes for the B subunit of cholera toxin. The recombinant strain SL3261 (named TT201) was obtained. Southern hybrid confirmed that the CTB gene was integrated into the chromosome of TT201. Western blot analysis demonstrated that the CTB gene was expressed stably in TT201. When administered orally to mice, the recombinant strain elicited a serum antibody response to CTB. The strain TT201 is a potentially bivalent vaccine candidate. PMID- 10465900 TI - Long-term behavioural effects of a repeated exposure to chlorphenvinphos in rats. AB - The aim of the present study was to investigate persistent neurobehavioural effects of repeated low-level exposure to chlorphenvinphos ((2-chloro-1-(2,4 dichlorophenyl) vinyl diethyl phosphate-CVP) in rats. The rats received 10 i.p. injections of CVP at daily doses of 0.5 mg/kg or 1.0 mg/kg (one injection/day, five days/week) which corresponded to 1/20 and 1/10 of LD50, respectively, for this species. In a part of the rats, cholinesterase (ChE) activity in blood (plasma and erythrocytes), and in the selected brain regions was determined at arbitrarily chosen time after the last exposure. The determinations showed that the level of ChE inhibition was dose-related, but the compartments studied differed in the magnitude of this effect. The differences in the level of ChE inhibition between the compartments were particularly evident in rats which had received CVP at the 1.0 mg/kg dose; in these animals 3 h after exposure the ChE activity in erythrocytes, plasma and the brain corresponded to 78%, 48% and 67 70%, respectively, of the control value. Enzyme activity returned to the control level after 14 days in plasma and after 35 days in erythrocytes. In rats receiving CVP at daily doses of 0.5 mg/kg, ChE activity in plasma was decreased by 40.8% and that in erythrocytes by 21.4% 3 h after the last exposure. The activity of ChE in plasma returned to the control level within four days and that in erythrocytes within 14 days. In these rats, in all the brain regions studied except brainstem, ChE activity was not reduced significantly. In rats selected for behavioural tests, the following behavioural aspects were investigated: response to novelty in an open field, acquisition and extinction of a one-way active avoidance response, and the magnitude and persistence of the footshock induced analgesia (hot-plate test). Testing in the open field was performed before the exposure and then 1, 3 and 6 weeks after the last exposure. The remaining tests were performed after the exposure. The interval between testing and the last CVP injection was sufficient for recovery of ChE activity. It has been found that in rats of both exposure groups the response to novelty in the open field, i.e. the increase in locomotor and exploratory activity in the presence of a new object, was reduced, albeit nonsignificantly, compared to the unexposed animals. In rats which received CVP at the dose of 1.0 mg/kg, acquisition of the one-way active avoidance response was facilitated. No differences between groups were found during extinction of this response. In the hot-plate test, in rats exposed repeatedly to 1.0 mg/kg CVP, the footshock induced increase in the latency of the paw-lick response to heat (54.5 degrees C) was stronger and more persistent than in the unexposed animals. The above results show that some neurobehavioural effects of exposure to organophosphorous (OP) compounds may be detected after a time sufficient for recovery of ChE activity. PMID- 10465901 TI - Effect of mustard gas on supraoxide dismutase activity and the level of malonyl dialdehyde: in vitro studies. AB - The investigation aimed at evaluating free supraoxide radicals generations and the degree of blood platelets cell membrane lipids peroxidation on the basis of supraoxide dismutase activity and the level of malonyl dialdehyde (MDA) after a 30-min exposure to mustard gas in vitro. The obtained results showed that blood platelets supraoxide dismutase activity and the MDA level are higher in comparison with the control groups. PMID- 10465902 TI - Effects of microwave irradiation on bacteria attached to the hospital white coats. AB - A test to sterilize pieces of cloths, which are the material of hospital white coats (doctors and nurses wears), by microwave irradiation in place of autoclaving was done using a commercial 2,450 MHz microwave oven. When pieces of cloths made of cotton (35%) and polyester (65%) were contaminated experimentally with Escherichia coli or Staphylococcus aureus and irradiated by microwave, the bacteria were killed quite rapidly according to almost first-order reaction kinetics. Only after a 20-sec irradiation, when the water content of cloths was decreased from the original 48% to 35%, the relative survivals of these bacteria fell to below 1% that of the non-irradiated control. The cloths were neither browned nor crisped, even after a 10-min irradiation of microwaves. PMID- 10465903 TI - An update of the frequency and type of diagnostic x-ray examinations in Poland. AB - This paper presents the results of a survey concerning x-ray diagnostic examinations in Poland, as the main source of radiation risk to the general population. The methods of data collection and processing were similar to that used for the previous national survey (1986). In 1995, 27,600,000 diagnostic x ray examinations were performed in Poland (6,200,000 more than in 1986). The number of examinations per 1,000 inhabitants increased from 572 to 715 during those nine years. The age and gender structure of examined patients is discussed extensively, the numbers for 1986 and 1995 are compared. Special attention is paid to the proportion of children and adolescents in the total number of patients examined. The obtained results are also compared with the numerical data concerning other countries. PMID- 10465904 TI - Some risk factors for non-Hodgkin's lymphoma. AB - Non-Hodgkin's lymphoma (NHL) has been subject to several epidemiological studies and various occupational and non-occupational exposures have been identified as determinants. The present study is a pooled analysis of two earlier methodologically similar case-referent studies encompassing 199 cases of NHL and 479 referents, all alive. Exposure information, mainly on occupational agents, was obtained by mailed questionnaires to the subjects. Exposure to white spirits, thinner, and aviation gasoline as well as work as a painter was connected with increased odds ratios, whereas no increased risk was noted for benzene. Farming was associated with a decreased odds ratio and exposure to phenoxy herbicides, wood preservatives, and work as a lumberjack showed increased odds ratios. Moreover, exposure to plastic and rubber chemicals and also contact with some kinds of pets appeared with increased odds ratios. Office employment and housework showed decreased odds ratios. This study indicates the importance of investigating exposures not occurring very frequently in the general population. Solvents were studied as a group of compounds but were also separated into various specific compounds. The present findings suggest that the carcinogenic property of solvents is not only related to the aromatic ones or to the occurrence of benzene contamination, but also to other types of compounds. PMID- 10465905 TI - Mortality in the cotton industry workers: results of a cohort study. AB - The cohort consisted of persons found on the payroll of one of the Lodz cotton plants in 1964-1993 who were employed in the plant for at least 10 years. Death risk by causes was analysed using standardised mortality ratios (SMRs) calculated by the person-years method. The general population of Poland was used as the reference. In all, 7892 people were observed. As of December 31, 1995, the follow up was completed for 7545 people (2852 men and 4693 women), i.e. the availability of the cohort was 95.6%. A total of 2069 deaths were recorded; the information on the cause of death was available for 97% of the subjects. In the male cohort, the level of the general mortality was the same as in the general population (SMR = 99). However, there was a significant increase in the number of deaths from diseases of the digestive system (SMR = 142) and larynx cancer (SMR = 188). The analysis of the results by production departments revealed in the weaving department significantly higher mortality from atherosclerosis (SMR = 141), peritoneal carcinoma (SMR = 1057) and melanoma (SMR = 677); and in the spinning department the increased risk of the hypertensive disease (SMR = 239), atherosclerosis (SMR = 175), and Hodgkin's disease (SMR = 768). Mortality in the female cohort was lower than that in the general population (SMR = 88). None of the disease groups or tumour sites caused statistically significant excess deaths either in the total cohort or in subcohorts selected according to departments. Special attention was paid to the chemical processing departments where chemicals used could contribute to the increased risk of death from cancer. Our analysis did not reveal any significant increase either in the total cohort of the workers employed in those departments or in the cohorts analyzed by duration of employment. Our results confirm the lower risk of lung cancer in the analysed group as compared with that in the general population. The numerous, but statistically insignificant increases in the incidence of malignant tumours at some specific sites detected in the subcohorts, distinguished according to the duration of employment or department, confirm the reported findings on the incidence of oral cavity, nose, throat, and larynx tumours among people exposed to harmful agents in the cotton industry. PMID- 10465906 TI - Evaluation of occupational exposure to infrasonic noise in Poland. AB - A short review of infrasound sources and effects on humans is presented. Polish standard PN-86/N-01338 and international standards ISO 7196:1995 and ISO 9612:1997 concerning the measuring techniques of infrasonic noise are described. The results of infrasonic noise measurements performed in the work environment in Poland are discussed. The study concerned the noise emitted by 124 different types of industrial machinery, appliances and means of transport. The measurements were made in typical working conditions with reference to Polish and international standards. The sound pressure levels exceeding Polish admissible values for: (a) workers' health protection were found in 5 (4.0%) cases, (b) ensuring proper conditions for performing basic functions in observational dispatcher cabins etc. in 77 (62.1%) cases; and (c) administration premises, design offices etc. in 92 (74.2%) cases. The admissible sound pressure levels for workers' health protection are in fact the permissible levels for hearing protection, however they do not correspond with the hearing threshold of infrasound the G-weighting characteristic is associated with. The hearing threshold of infrasound (G86 curve) was exceeded in 66.9% of all the industrial machinery and means of transport under study. PMID- 10465907 TI - Tinnitus and impulse noise-induced hearing loss in drop-forge operators. AB - Tinnitus is frequently accompanied by noise-induced hearing losses, and perhaps as it is assumed--particularly with those arising from exposure to impulsive types of noise. In order to explain it and to estimate a prevalence of tinnitus in the industrial, impulse noise workers the group of 261 drop-forge operators exposed to impulses with peak levels of 135 dB versus 169 age-matched controls was subjected to otological and audiometric examination, and then the complaints for tinnitus in both groups have been analysed. The prevalence of tinnitus, highest in operators with longer exposure duration (> 10 years) was found in 184 individuals (70.4%) versus 6 (3.5%) in controls. The findings closely corresponded with the degree of impulse noise-induced hearing loss. As the maximum audiometric notch was mostly localised at 6 kHz and rarely at 4 kHz, consequently the approximate pitch of the tinnitus was related to the frequencies where hearing was most affected. It is concluded that impulse noise-induced tinnitus may be sometimes more severe in its effects than is hearing loss, thus it creates an additional reason for strict hearing conservation programmes. PMID- 10465908 TI - The role of otoacoustic emissions in screening and evaluation of noise damage. AB - Otoacoustic emissions have been shown to be extremely useful in screening for hearing impairment in babies. This review considers the current evidence available with regard to the role of otoacoustic emissions in the screening for noise induced cochlear damage. A number of studies indicate that otoacoustic emissions provide an indication of cochlear damage prior to any change in the pure tone audiometric threshold. There is also some evidence that the medial efferent system evaluation using contralateral sound activated suppression of emissions indicates dysfunction after noise exposure. Increased variability of spontaneous emissions has also been shown to be associated with the presence of tinnitus in various aetiologies. PMID- 10465909 TI - Antibiotics and sacred cows. PMID- 10465910 TI - Antibiotics and sacred cows. PMID- 10465911 TI - Antibiotics and sacred cows. PMID- 10465912 TI - Antibiotics and sacred cows. PMID- 10465913 TI - Amalgamation: choice or necessity? PMID- 10465914 TI - Dalacin C advertisement. PMID- 10465915 TI - Considerations re: sweeteners in medication. Canadian Dental Association. PMID- 10465916 TI - Considerations re: use of latex in dentistry. Canadian Dental Association. PMID- 10465917 TI - Using simulation to evaluate clinical competence after impairment. AB - It is important for individual dentists and the profession to have access to a process for evaluating the clinical competence of practitioners who are professionally impaired as a result of an accident or a medical disability. No common standards for such evaluations currently exist, however, as demand for this type of assessment is still rare. This article reviews the evaluative approach taken by a team of experienced dental educators in examining three dentists who suffered from medical disabilities. An attempt was made to standardize the evaluation process by using clinical simulation to create an environment that would be comfortable for the dentists and acceptable to the lawyers and the insurance companies. Following evaluation, recommendations on individual competence were made, contributing to a faster resolution of legal and insurance issues. PMID- 10465918 TI - Detecting child abuse and neglect--are dentists doing enough? AB - Dental health professionals continue to under-report child abuse, despite growing awareness of their potential role in detecting this crime. This article presents an overview of child abuse and neglect and outlines the indicators that may alert dental professionals to possible maltreatment of child patients. Documentation protocols are also provided to aid in reporting child abuse identified in the dental office. PMID- 10465919 TI - A plea for a dental research institute. PMID- 10465920 TI - [Guided tissue regeneration in endodontics. Part 1]. AB - Over the past twenty years, Guided Tissue Regeneration (GTR) has become an accepted technique for the treatment of tissue loss consequent to periodontal disease. The principles of GTR have recently begun to be employed in endodontics since pulpal necrosis often leads to periodontal tissue loss as a result of the leakage of noxious irritants through apical foramina and accessory canals. This treatment modality is still in its early days for the management of endo-perio lesions and is most appropriately used in specific types of cases. This article, the first of a two-part series, presents a review of the literature about the mechanisms of action of GTR and discusses some of the products used to bring about GTR. PMID- 10465921 TI - Determination of the sensitivity and specificity of PCR assays using different target dnas for the detection of Mycobacterium tuberculosis. AB - To establish a sensitive, specific and reproducible PCR assay for the detection of Mycobacterium tuberculosis, we evaluated three target DNAs: IS6110, 65 kDa heat shock protein gene; and mtp40 genomic fragment. We purified genomic DNA from 15 mycobacterial strains including four M. tuberculosis isolates, four M. bovis BCG isolates, and one of each for M. fortuitum, M. avium, M. intracellulare, M. szulgai, M. scrofulaceum, M. chelonei, and M. gordonae from the culture and used them as the template DNA. We studied 3 primer sets for IS6110, 2 primer sets for 65 kDa, and 3 primer sets for mtp40. Depending on the assay, these primer sets were used in the single-step PCR and/or nested PCR. The PCR assay targeting the 65 kDa protein gene could detect all of the tested mycobacterial strains, whereas targeting the IS6110 sequence resulted in detection of only M. tuberculosis and M. bovis BCG. Furthermore, targeting the mtp40 genomic fragment could be used to distinguish M. tuberculosis from M. bovis BCG. Using a nested PCR assay with primer sets specifically targeting the IS6110 or 65 kDa, we have been able to detect single copy M. tuberculosis genomic DNA. When the mtp40 genomic fragment was used as the target DNA, the sensitivity of detection was 10 copies of M. tuberculosis genomic DNA. This assay was demonstrated to have good sensitivity and specificity for detection and discrimination of mycobacterial species, and could be used in analyzing the clinical samples with low copy number infections such as the cerebrospinal fluid from the patient with tuberculous meningitis. PMID- 10465922 TI - Acinetobacter baumannii bloodstream infection: clinical features and antimicrobial susceptibilities of isolates. AB - The number of nosocomial infections caused by Acinetobacter baumannii has increased in recent years. The purposes of this study are to discover the risk factors of transmission to prevent the nosocomial infection of A. baumannii. We retrospectively studied 36 patients with A. baumannii bacteremia at China Medical College Hospital from January 1996 to December 1997. There were 23 males and 13 females. All bacteremia were acquired nosocomially. Malignancy (n = 8) and intracranial hemorrhage (n = 6) were the most common underlying diseases. Only one patient on arterial line disclosed intraarterial catheter-related A. baumannii bacteremia and 3 patients had evidence of A. baumannii pneumonia. Twenty-one patients (58%) had central venous catheters in place at the onset of bacteremia, but none was proven to be catheter-related infection. There were 32 patients (89%) with unknown portal of entry. Multivariate logistic regression analysis revealed that potential risk factors related to A. baumannii bacteremia were prior antimicrobial therapy (P < 0.05). The most common clinical features of A. baumannii bacteremia were, in descending order, fever, leukocytosis, thrombocytopenia and hypotension. Eleven patients (30.6%) died directly from A. baumannii bacteremia. All isolates were resistant to ampicillin, cephalothin, cefonicid and moxalactam. The most alarming evidence was that 19% of isolates showed resistance to imipenem. Our findings emphasized that A. baumannii bacteremia had the following characteristics: usually acquired nosocomially, unknown portal of entry, and high multiresistance, especially the increasing resistance rate to imipenem. Imipenem must be reserved as a last-line agent to treat A. baumannii infections, so we want to suggest that the treatment of choice for A. baumannii is gentamicin, amikacin or ceftazidime. PMID- 10465923 TI - Comparison of necrotic characteristics and benefits between 50% acetic acid and pure ethanol in local hepatic injection: a study in rats. AB - This study compares the characteristics of acetic acid and pure ethanol in local hepatic injection and identifies the possible benefits from acetic acid by an animal model. We injected 0.03 ml of pure ethanol and 0.01 ml of 50% acetic acid separately into different sites of the same liver. The shape and diameter of the lesions were recorded at 0 min, 5 min and 15 min, and 5 lesions were also reviewed one day later. The lesions induced by pure ethanol were irregular, whereas those induced by 50% acetic acid were round or oval. The diameter of lesions induced by 50% acetic acid was 2.62 +/- 0.58 mm initially and increased to 4.50 +/- 0.84 mm and 5.25 +/- 0.75 mm at 5 min and 15 min, (5 min compared with 0 min, p < 0.0001; 15 min compared with 5 min, p < 0.001) respectively. The diameters of lesions induced by pure ethanol did not increase. The diffusion diameter at 5 min and 15 min were 1.88 mm and 2.63 mm respectively. Furthermore, viable liver cells were occasionally found in two lesions induced by pure ethanol. Acetic acid had a more homogeneous distribution and better infiltrating ability than pure ethanol and may be superior to pure ethanol in local injection therapy of HCC. But more clinical trials in the treatment HCC should be carried out. PMID- 10465924 TI - Comparison of subcutaneous hydromorphone with intramuscular meperidine for immediate postoperative analgesia. AB - Intramuscular (i.m.) injection with meperidine is the most common analgesic approach to treat postoperative pain in Taiwan. Hydromorphone (Dilaudid) can provide very potent and rapid analgesic effect through subcutaneous (s.c.) injection. Although hydromorphone is widely used in North America, no study has compared the analgesic efficacy, side effect profiles and patients' satisfaction with the method of injection of hydromorphone s.c. and meperidine i.m. for the immediate post-operative analgesia. In this randomized and double-blind study, 60 female patients scheduled for abdominal total hysterectomy were treated either with 1 mg hydromorphone s.c. (n = 30) or 50 mg meperidine i.m. (n = 30) when they regained consciousness and asked for analgesic treatment in the recovery room. Visual analogue score (VAS) of wound pain was obtained at 0, 10 and 30 min after injection by a blinded observer. The occurrence and severity of nausea, vomiting, dizziness, drowsiness, flatus passage and respiratory depression were recorded. Post-operative analgesia in the ward was maintained by patient-controlled analgesia (PCA) with intravenous morphine. Time to first PCA demand, the number of demands, delivery, delivery/demand ratio and 24 h morphine consumption were documented. We found that VAS was reduced at 10 min and, to a greater extent, at 30 min postinjection in both groups but with no significant difference between the two groups. The occurrence and severity of side effect profiles were similar in both groups except that dizziness was more frequently observed after meperidine injection. Delivery, demand, delivery/demand ratio and 24 hr morphine consumption by PCA were not significantly different between the two groups. Time to first PCA trigger was also similar. Patients receiving hydromorphone s.c. injection exhibited higher satisfactory score than those receiving meperidine i.m. injection. Hydromorphone 1 mg, injected subcutaneously, was as effective as intramuscular meperidine 50 mg while permitting more favorable injection technique and fewer side effects. We suggest that subcutaneous hydromorphone is a good alternative to intramuscular meperidine for postoperative analgesia in the recovery room. PMID- 10465925 TI - An investigation into the opinions of nursing faculty toward the RN/RPN licensure exams. AB - The purpose of this study was to investigate opinions on the RN/LPN licensure exams held by nursing faculty teaching at vocational nursing schools, community nursing colleges and full universities. The research process consisted of questionnaire construction and major study phases. In phase I, faculty members were chosen from one community college and given tape recorded interviews which were then transcribed. Interview data were analyzed and yielded the inventories of the opinion and attitude towards the RN/LPN licensure exams. Kendall's Coefficient of Concordance was used to test the content validity and yielded w = .18 (p < .01) for opinion and w = .42 (p < .01) for attitude. Cronbach's Alpha was used to test internal consistency and yielded 0.84 and 0.69 respectively. In phase II, nursing faculty members were chosen from all levels of institutions offering nursing degrees in Taiwan. The responses for item opinion towards the RN/LPN licensure exams ranged from 22-65 with a mean score of 47.61; while responses for attitude towards item constructors of the licensure exams ranged from 11-29 with a mean score of 19.16. The attitude towards the RN/LPN exam constructors had significant negative correlations with age, clinical experience, and teaching experience, but had significant positive correlation with opinion towards the items on the RN/LPN licensure exams. There was significant negative correlation between the opinion towards the items on RN/LPN exams and clinical experience (p < .05). The faculty members with experience in item construction for the RN/LPN licensure exams showed a significantly greater negative opinion in the attitude towards the RN/LPN licensure exam constructors compared to faculty without experience in item construction (p < .01). A significant difference in scores of attitude toward the RN/LPN licensure exams was found with different faculty educational levels and different teaching positions within the various types of institutions (p < .01). Further studies to engage in item analysis for the RN/LPN exam are recommended. PMID- 10465926 TI - Acute painful oculomotor nerve paresis caused by pituitary apoplexy--a case report. AB - In a typical case of pituitary apoplexy, a patient, who may or may not be known to harbor a pituitary adenoma, suddenly develops a severe headache. The headache may be retro-orbital, frontal, frontotemporal, or diffuse and may be associated with neck stiffness, neck pain, or both. Ophthalmoplegia may develop within a few hours after the onset of headache. Here we report a rare case of one middle-aged female with pituitary apoplexy initially presenting with acute onset of pupil involved third cranial nerve palsy, headache and peri-ocular pain. Emergent neuroimaging revealed pituitary apoplexy and immediate intravenous corticosteroid was given and third nerve paresis was improved thereafter. Definite tumor removal was done smoothly after steroid treatment and complete recovery of ophthalmoplegia was noted 2 weeks after operation. PMID- 10465927 TI - Surgical excision of the tendon xanthoma in familial hypercholesterolemia--a case report. AB - Familial hypercholesterolemia is an autosomal dominant disorder characterized by increased low-density lipoprotein cholesterol, premature atherosclerosis and tendon xanthomas. Genetic studies reveal familial hypercholesterolemia to be a dysfunction of LDL receptor gene on cell surface. Recently various mutations in the LDL receptor gene have been reported. When DNA method is not available, the occurrence of tendon xanthomas, an isolated elevation of plasma cholesterol, with a normal concentration of plasma triglycerides virtually establishes the diagnosis of familial hypercholesterolemia. In this report, a 42-year-old male had tendon xanthoma at extensor surface of metacarpophalangeal joint of his right hand, olecranon of the left elbow and both knees, and Achilles tendons. The tendon xanthoma was excised for cosmetic reasons, and the wound healing was slower than average in this case. We suggest that before suture removal, wound healing must be complete. It is important that the hand surgeon recognize that tendon xanthoma is a physical sign of a potentially life-threatening disorder to the patient as well as his family, and that this disorder may respond favorably to early examination and management. PMID- 10465928 TI - Complementary role of dual isotope in non-bacteriuric renal infection--a case report. AB - To detect non-bacteriuric renal infection in a diabetic patient, though difficult but is very important because early diagnosis and early treatment can prevent later complications such as renal abscess, renal hypertension or even end stage renal disease. Herein, we presented a case of diabetic patient with septicemia whose urine culture and renal ultrasonography were negative initially. By using a combination of dual isotope images and single photon emission computed tomography technique, an infectious lesion in the upper pole of left kidney was revealed, which was identified as acute focal bacterial nephritis by computed tomography four days later. This case report showed that 67Ga plus 99mTc dimercaptosuccinic acid images are useful in patients under clinical suspicion of renal infection, especially for those with negative urine analysis and/or urine culture initially. PMID- 10465929 TI - Identifiability of parameters in the Yakovlev-Polig model of carcinogenesis. AB - The paper discusses the problem of identifiability for two versions of a two stage model of carcinogenesis recently introduced by Yakovlev and Polig. In this model, cell killing is allowed to compete with tumor promotion. In the first version of the Yakovlev-Polig model, which is referred to as Model 1, cell killing starts immediately after a carcinogen is administered. In the second version, called Model 2, it is assumed that a cell may be killed only after the process of initiation has been completed. The two versions of the Yakovlev-Polig model suggest explicit formulas for the distribution of time to tumor onset (that is, appearance of the first malignant clonogenic cell) counted from the initial moment of the exposure to a carcinogen. A model of carcinogenesis is identifiable if the set of all model parameters is uniquely determined by the distribution of time to tumor onset. It is shown that, under a natural necessary condition of overlap of supports of the dose-rate function h and the promotion time distributions from a family F, Model 1 is identifiable in the family F for many practically important functions h. In particular, this is the case for a simple model of spontaneous carcinogenesis (h = 1) and for a class of piecewise constant dose-rate functions h with arbitrary family F. Also, this holds for the family of gamma distributions and h supported on an interval and non-vanishing in the interior of this interval. More restrictions need to be imposed on the dose-rate function and the family of promotion time distributions for Model 2 to be identifiable. In particular, for h = 1, Model 2 turns out to be non-identifiable even in the family of gamma distributions. PMID- 10465930 TI - A comparison of foraging strategies in a patchy environment. AB - In this paper we compare foraging strategies that might be used by predators seeking prey in a patchy environment. The strategies differ in the extent to which predators aggregate in response to prey density. The approach to the comparison is suggested by the idea of evolutionarily stable strategies. A strategy is said to be evolutionarily stable if it cannot be invaded by another strategy. Thus we examine scenarios where a small number of individuals using one strategy are introduced into a situation where a large number of individuals using the other strategy are already present. However, our foraging models do not explicitly incorporate predator population dynamics, so we use net energy uptake as a surrogate for reproductive fitness. In cases where all of the patches visited by predators sustain prey populations, we find that for any pair of strategies one of them will have a higher net energy uptake than the other whether it is the resident or the introduced strain. However, which one is higher will typically depend on the total predator population, which is determined by the resident strain. If the predators leave prey densities high, the more aggregative strain will have the advantage. If the predators reduce prey densities to low levels the less aggregative strain will have the advantage. In cases where one strain of predators aggregates in response to prey density and the other does not, then there might be patches which do not contain prey but do contain (non-aggregating) predators. In those cases, there is the possibility that whichever strategy is used by the introduced strain will yield a higher energy uptake than that used by the resident strain. This suggests that if some patches are empty of prey then aggregative and non-aggregative strategies may be able to coexist. PMID- 10465931 TI - Existence and global attractivity of positive periodic solutions of periodic n species Lotka-Volterra competition systems with several deviating arguments. AB - In this paper, we study the existence and global attractivity of positive periodic solutions of periodic n-species Lotka-Volterra competition systems. By using the method of coincidence degree and Lyapunov functional, a set of easily verifiable sufficient conditions are derived for the existence of at least one strictly positive (componentwise) periodic solution of periodic n-species Lotka Volterra competition systems with several deviating arguments and the existence of a unique globally asymptotically stable periodic solution with strictly positive components of periodic n-species Lotka-Volterra competition system with several delays. Some new results are obtained. As an application, we also examine some special cases of the system we considered, which have been studied extensively in the literature. Some known results are improved and generalized. PMID- 10465932 TI - Relationships between antiviral treatment effects and biphasic viral decay rates in modeling HIV dynamics. AB - Recently, potent combination antiviral therapies consisting of reverse transcriptase inhibitor (RTI) drugs and protease inhibitor (PI) drugs, have been developed which can rapidly suppress HIV below the limit of detection. Two phases of plasma viral decay after initiation of treatment have been observed in clinical studies. Some researchers have suggested that the viral decay rates may reflect the potency (efficacy) of antiviral therapies. In this paper we model the effect of RTI drugs and PI drugs as inhibition rates of cell infection and infectious virus production, respectively, based on the biological mechanisms of these two different types of drugs. Through rigorous mathematical derivation, we show that the two viral decay rates are monotone functions of the treatment effects of these antiviral therapies. We derive approximation formulas for the relationships between viral decay rates and treatment effects. Computer simulations show that the approximation formulas approximate the true values very well. These formulas may be used to study what factors really affect the viral decay rates. The results in this paper provide a theoretical justification for using both viral decay rates for evaluation of the treatment efficacy of antiviral therapies. PMID- 10465933 TI - Structured models for heterosexual disease transmission. AB - An SIS model for a heterosexually transmitted disease with core and non-core compartments and a generalized recovery function P(t) is analyzed. It exhibits R0 threshold behavior and leads to discussions of stability with respect to choice of P(t), and of the effects of allowing recruitment between core and non-core groups. PMID- 10465934 TI - [Progress on the diagnosis and therapy of anemia]. PMID- 10465935 TI - [Progress on the diagnosis and therapy of anemia--cell differentiation of erythroblasts and hematopoiesis factors]. PMID- 10465936 TI - [Diagnostic study in anemia]. PMID- 10465937 TI - [Diagnosis and therapy of hypochromic anemia]. PMID- 10465938 TI - [Diagnosis and therapy of megaloblastic anemia]. PMID- 10465939 TI - [Diagnosis and therapy of aplastic anemia]. PMID- 10465940 TI - [Diagnosis and therapy of pure red cell aplasia]. PMID- 10465941 TI - [Diagnosis and therapy of myelodysplastic syndrome]. PMID- 10465942 TI - [Diagnosis and therapy of hemolytic anemia--genetic analysis and membrane protein deficiency in hereditary spherocytosis]. PMID- 10465943 TI - [Diagnosis and therapy of hemoglobinopathies]. PMID- 10465944 TI - [Diagnosis and therapy of autoimmune hemolytic anemia]. PMID- 10465945 TI - [Diagnosis and therapy of paroxysmal nocturnal hemoglobinuria]. PMID- 10465946 TI - [Diagnosis and therapy of renal anemia]. PMID- 10465948 TI - [Diagnosis and therapy of drug-induced hematopoietic disorders]. PMID- 10465947 TI - [diagnosis and therapy of anemia of chronic disorders]. PMID- 10465949 TI - [Diagnosis and therapy of anemia in the elderly]. PMID- 10465950 TI - [Diagnosis and therapy of pediatric patients with anemia]. PMID- 10465951 TI - [Progress on diagnostic and therapeutic study of anemia (discussion)]. PMID- 10465952 TI - [Cases of pseudothrombocytopenia in a mother and the newborn child]. PMID- 10465953 TI - [Case of thymoma complicated with rheumatoid arthritis]. PMID- 10465954 TI - [Chemotherapy of endocarditis due to Candida glabrata]. PMID- 10465955 TI - [Case of Creutzfeldt-Jakob syndrome presenting homonymous field defect in its early stage]. PMID- 10465956 TI - [Case of melioidosis associated with acute empyema and cellulitis of the leg]. PMID- 10465957 TI - [Organogenesis and differentiation of the cardiovascular system]. PMID- 10465958 TI - [NK/T cell neoplasms]. PMID- 10465959 TI - [Channelopathies]. PMID- 10465960 TI - [Parathyroid gland diseases and calcium metabolism disorders]. PMID- 10465961 TI - [Diagnosis of hypercalcemia]. PMID- 10465962 TI - [Diagnosis of hypocalcemia]. PMID- 10465963 TI - [Diagnosis of phosphorus metabolism disorders]. PMID- 10465964 TI - [Diagnosis of magnesium metabolism disorders]. PMID- 10465965 TI - [Physiopathology and therapy of primary hyperparathyroidism]. PMID- 10465966 TI - [Physiopathology and therapy of hypercalcemia associated with malignancies]. PMID- 10465967 TI - [Physiopathology and therapy of familial hypocalciuric hypercalcemia]. PMID- 10465968 TI - [Physiopathology and therapy of multiple endocrine neoplasia type 1 and type 2A]. PMID- 10465969 TI - [Physiopathology and therapy of hypercalcemia crisis]. PMID- 10465971 TI - [Physiopathology and therapy of pseudohypoparathyroidism and pseudopseudohypoparathyroidism]. PMID- 10465970 TI - [Physiopathology and therapy of idiopathic and other hypoparathyroidism]. PMID- 10465972 TI - [Physiopathology and therapy of renal osteodystrophy and secondary hyperparathyroidism]. PMID- 10465973 TI - [Physiopathology and therapy of rickets and osteomalacia]. PMID- 10465974 TI - [Physiopathology and therapy of osteoporosis]. PMID- 10465975 TI - [Recent progress on the study of calcium-metabolism regulating factors- parathyroid hormone and calcium-sensing receptor]. PMID- 10465976 TI - [Recent progress on the study of calcium-metabolism regulating factors- parathyroid hormone-related protein and its function]. PMID- 10465977 TI - [Recent progress on the study of calcium-metabolism regulating factors- biosynthesis pathway of vitamin D and its function]. PMID- 10465978 TI - [Recent progress on the study of calcium-metabolism regulating factors- parathyroid gland hyperplasia and molecular biology of neoplasms]. PMID- 10465979 TI - [Recent progress on the study of calcium-metabolism regulating factors--oncogenic osteomalacia]. PMID- 10465980 TI - [Topics on hypercalcemia and hypocalcemia (discussion)]. PMID- 10465982 TI - [Case of diabetes mellitus with mitochondrial cardiomyopathy]. PMID- 10465981 TI - [Case of systemic Castleman's disease with hypothyroidism]. PMID- 10465983 TI - [Case of peritoneal tuberculosis associated with progressive genital tuberculosis]. PMID- 10465985 TI - [Case of massive osteolysis]. PMID- 10465984 TI - [Long-time survival case of glucagonoma metastatic to the liver treated with TAE therapy]. PMID- 10465986 TI - [Prevention of cross infections in immunocompromised hosts--the theory and reality]. PMID- 10465987 TI - [Gentic study of collagen diseases]. PMID- 10465988 TI - [Magnetic resonance cholangiopancreatography (MRCP)]. PMID- 10465989 TI - [Comparison between the poisoning severity score and specific grading scales used at the Department of Clinical Toxicology in Krakow]. AB - The aim of the present study has been to assess the concordance in severity grading when using the Poisoning Severity Score (PSS), proposed by IPCS/EC/EAPCCT vs. some specific grading scales. Cases of acute poisoning admitted to the Department of Clinical Toxicology in Krakow during months January, March, May, July, August and October 1998 (n = 820) were evaluated. Severity grading was performed in all cases using both the PSS and special grading scales developed by the poisons centre in Krakow. The PSS is assessing severity on the basis of observed clinical signs and symptoms (at their maximum), but does not take into account potential risks or plasma/serum concentrations. The Krakow scales include both clinical symptoms on admission and results of toxicological analyses. Ethanol (39.4%), drugs (36.2%) and carbon monoxide (7.9%) were the most commonly involved toxic agents. Overall concordance between the PSS and the Krakow scales was at hand in 596 cases (72.7%). The lack of concordance was most evident for ethanol and carbon monoxide cases--the PSS generally giving a lower grade. The discordance was less pronounced for grade 3 (life-threatening) poisonings. Significant number of ethanol poisoning in Krakow centre require medicolegal certification thus ethanol concentration (blood or expired air) is considered while evaluating the poisoning severity. Moreover majority (more than 70%) of acutely ethanol poisoned patients treated at the Department are chronic alcoholics so clinical evidence of intoxication may be minimal at blood ethanol levels higher than 3.0 g/L. For ethanol intoxication the blood ethanol concentration was considered in the Krakow scale but not in the PSS, and when the blood concentrations were disregarded and clinical symptoms only were evaluated the concordance improved between the different systems from 72.7% to 92.3%. Thereby also the overall concordance in the study increased to 82.9%. The less satisfying concordance for carbon monoxide cases was mainly caused by discrepancies in the evaluation of neurological symptoms. Also the blood lactate concentration, COHb level, duration of exposure and patient's age are considered in the Krakow scale but not in PSS. The PSS is intended as a general scheme for grading severity of acute poisoning. In this study an acceptable concordance between the PSS and some locally developed grading scales was at hand in the majority of cases, but it seems that for specific poisons, like carbon monoxide, some modifications and additional criteria may be justified. Further studies to test the reliability of the PSS are encouraged. PMID- 10465990 TI - Acute poisonings with chemical compounds among adolescent and adult inhabitants of Krakow in the year 1997. AB - The pattern of adolescent and adult poisonings in Krakow is presented on the basis of the data collected by the Poison Information Centre of the Department of Clinical Toxicology Collegium Medicum of the Jagiellonian University in 1997. This analysis includes 3472 people treated at the Department of Clinical Toxicology and 118 poisoned people who were not given any treatment and died at the scene of the accident. The group of hospitalised persons consisted of 2359 (67.9%) men and 1113 (32.1%) women, and the group of people who died at the scene of the accident consisted of 97 (82.2%) men and of 21 (17.8%) women. The overall coefficient of poisonings in 1997 was 46.9; for men--67.8 and 28.3 for women. Ethanol (39.7%) followed by drugs (20.2%), drugs + ethanol (6.7%), carbon monoxide (6.2%) and drugs of abuse (4.4%) were the most common cause of acute poisonings. The mortality rate of the cases treated was low (0.43%), but while including those people who were not given any treatment and died at the scene of the accident, mostly due to ethanol, carbon monoxide, and drugs poisoning the fatality index rose up to 3.7%. PMID- 10465991 TI - Frequency and clinical course of acute carbon monoxide poisoning in relation to the kind of emission gas. AB - The conversion of domestic supply from illuminating to natural gas in 1975 in Krakow was expected to diminish, or even eliminate the threat of CO intoxication. The aim of the study was to compare the frequency and clinical course of CO poisonings (early or delayed medical complication or death) regarding the source of CO (illuminating or natural gas). Under analysis there were adult inhabitants of Krakow treated at the Department of Clinical Toxicology from 1972 to 1974 when the main source of CO was illuminating gas and those treated from 1994 to 1996 i.e. after the conversion of domestic gas supply from illuminating to natural gas. All the cases of people who died at the scene of the accident prior to any treatment were also included. The frequency of acute poisonings among the inhabitants of Krakow was expressed by the incidence rate per 10,000 inhabitants with regard to age and sex. The severity of CO poisoning was estimated considering age, duration of exposure, carboxyhemoglobin (COHb) level, and the neurological state of patients on admission to the Clinic. Significantly higher frequency of suicidal poisonings was noted in the group of patients poisoned with illuminating gas compared to those poisoned with CO produced by incomplete combustion of natural gas (p < 0.001). No significant differences were noted considering the age of patients hospitalised in both periods. Duration of exposure was significantly longer and carboxyhemoglobin level was significantly higher in the group of patients poisoned with illuminating compared to natural gas. Significantly higher frequency of severe poisonings was noted in the group of patients intoxicated with illuminating gas compared to those intoxicated with natural gas. The frequency of acute CO poisonings in the population of Krakow determined by the incidence rate per 10,000 inhabitants was approximated to 2.4 in all the years analysed. The conversion of domestic supply from illuminating to natural gas significantly decreased the mortality rate. No differences in the number of patients suffering from medical complications were found between the two study periods. A higher incidence of late sequalae (psychoorganic and depressive syndromes) was found in the group of patients poisoned with CO produced by incomplete combustion of natural gas compared to illuminating gas. PMID- 10465992 TI - [Social conditions of suicidal poisonings]. AB - The paper presents the results of a study of social conditioning of suicidal poisonings conducted in the years 1994-1996. The study was of the case-control study type and covered 542 intoxicated patients treated in the Toxicology Department of the Medical Faculty of the Jagiellonian University in Krakow. The studied population consisted of 323 patients poisoned in suicidal attempts (the studied group) and 219 accidentally poisoned (the control group). The objective of the study was to determine social conditioning of suicidal poisonings. It has been found that the risk of suicidal attempt was 2.3 times higher among divorced or separated people; 2.2 times higher among those, who had recently lost their jobs; 2.4 times higher among people having financial problems while nearly twice lower among individuals having more children. PMID- 10465993 TI - The comparison of respiratory resistance and occlusion pressure in alcohol dependent patients who complain or not of the respiratory system disorders. AB - Alcohol abusers often complain of respiratory symptoms. The aim of the study was to analyse the respiratory pattern parameters and occlusion pressure in chronic alcoholics who complained of respiratory symptoms vs. those who did not suffer from any symptoms before the admission. Also dynamics in changes of spirometry parameters and respiratory pattern parameters during controlled, absolute abstinence while hospitalisation in the Detoxification Unit of the Department of Clinical Toxicology was evaluated. There were 124 study patients: 84 ethanol dependent patients and 40 healthy subject not dependent on ethanol (control group). Ethanol dependency was diagnosed using ICD-10 criteria. The questionnaire according to Fletcher was gathered for each of the subject. The positive results of Fletcher questionnaire (chronic cough + chronic expectoration) was obtained in 43 of the ethanol abusers-group I. The rest of abusers who did not complain of any respiratory symptoms was included to the group II. Respiratory tract resistance was significant the differential factor between the patients with positive results of Fletcher questionnaire and patients who did not suffer any symptoms before admission. Significant differences in occlusion pressure values, were noted between the group of abusers with and without the respiratory symptoms both in the first and control examination. PMID- 10465994 TI - Assessment of nutritional state of alcohol abusers and drug poisoned patients. AB - Beside the primary illness and environmental factors also alcohol addiction, similarly to another kind of dependency, is being considered as the causative factor of nutritional disorders. Alcohol and drug dependent people due to distorted eating behaviour are predisposed not only to eating disorders (anorexia, bulimia) but also to eating abuse (obesity, overweight, habitual eating). The aim of the study was to assess the nutritional state of alcohol abusers and drug poisoned patients. 113 patients treated at the Department of Clinical Toxicology in Krakow including 65 chronic alcoholics (85% men) and 48 drug poisoned patients (75% young women) were examined anthropometricaly. The measures were performed twice: on admission and on discharge from the Clinic. Nutritional state was unsatisfactory in 76.1% of examined patients and malnutrition or risk of malnutrition was more prevalent than overnutrition. Proportions of subjects qualified to specific groups of nutriture were different in alcohol abusers compared to drug poisoned patients. Also etiology of malnutrition was different. Irregular lifestyle and improper nutrition mode in the group of alcohol abusers (mainly men) and a phenomenon of "slimness terror" in the group of young woman with incidence of suicide attempt should be understood as significant factors of malnutrition. There was no detectable change in nutriture within the nine days long hospitalisation. Only slight, not significant, increase in values of anthropometric parameters was observed in the group of alcohol dependent patients. PMID- 10465996 TI - [Methods of poison elimination--controversies and actual views about their use]. AB - This paper presents controversies and current ideas about application of poison elimination methods in acute poisonings. Indications and contra-indications for single dose of activated charcoal, gastric lavage, cathartics and whole bowel irrigation are discussed. Rationale and indications for forced diuresis, repeated doses of activated charcoal, hemodialysis and hemoperfusion, enhanced elimination methods, are described. PMID- 10465995 TI - [Suicidal poisoning with tricyclic antidepressants]. AB - In the period from 1989 to 1998, 98 patients with suicidal tricyclic antidepressants poisoning were treated, including 47 men and 51 women from 16 to 74 (mean 35) years old. Thirty persons were poisoned by tricyclic antidepressants, in 68 remaining cases intoxications were mixed eg. including tricyclic antidepressants and benzodiazepines, phenothiazines, barbiturates, alcohol, amizepine, salicylates. The main causes of suicides were various kinds of depression (66%) as well as psychopathy and sociopathy (18%). Twenty patients attempted repeatedly to commit suicide. These were mainly patients suffering from endogenous depression, psychopathy and schizophrenia. In the examined group five patients died (5%) but three women out of them were aged, suffered from congestive heart failure and were poisoned with tricyclic antidepressants, benzodiazepines and barbiturates. PMID- 10465997 TI - [Amanita phalloides poisoning--diagnosis, clinical course, treatment]. AB - This study presents a general review of current opinion on Amanita phalloides and other toxic Amanita species poisonings, clinical symptoms, elementary principles of mycological, clinical and laboratory diagnostics, and treatment. Early diagnosis, centralization of the treatment have been particularly emphasized. PMID- 10465998 TI - [The role of laboratory methods in clinical and forensic toxicology]. AB - In the present work problems of differences between laboratory methods used in contemporary toxicological clinical and forensic analysis. The significance of instrumental methods as a factor quarantined proper level of scientific investigation has been underlined. A role of an analyst in clinical laboratory and an expert in toxicological forensic areas has been discussed. PMID- 10465999 TI - Suicidal paracetamol poisoning of a pregnant woman just before a delivery. AB - The aim of the paper is to present a case of self-poisoning with paracetamol, overdosed just before a delivery. A 21-year-old woman was admitted to Obstetric and Gynecology Ward of local hospital in the second stage of physiological delivery, more than 6 hours after she had ingested 19 g of acetaminophen for self poisoning. She delivered a normal infant weighing 3520 g who had Apgar scores of 10, and then both infant and mother were sent in an emergency ambulance to the nearest poison centre. Blood samples for toxicological examination were taken on admission to toxicological intensive care unit i.e. 11 hours post maternal ingestion. Acetaminophen levels of both patients were above the acetaminophen overdose nomogram line and the antidote treatment, i.v. N-acetylcysteine was administered according to the protocol: the mother within 11 hours post-ingestion and approximately 4 hours after a delivery; the neonate within 11 hours post maternal ingestion and 4 hours of life. Higher paracetamol concentration in the blood of infant compared to the mother's was noted in the first and then control toxicological examination performed within 35 hours post maternal ingestion. Peak maternal aspartate aminotransferase (AST) activity was 326 U/L within 35 hours and alanine aminotransferase (ALT) activity was 262 U/L within 56 hours post ingestion. The highest neonatal enzyme activity was noted within 11 hours post maternal ingestion of paracetamol, and the elevation was not high. Except moderate anaemia in the mother, no clinical or biochemical symptoms of renal, cardiovascular or CNS injury were stated in the mother or infant. Normalisation in the maternal enzymes activity was stated within 226 hours, while in the neonatal within 58 hours post maternal ingestion. The woman recovered without sequelae and was discharged from hospital on the 11th day following paracetamol overdosing. No evidence of the liver injury was found in the infant either. PMID- 10466000 TI - [Acute, oral poisoning with hydrogen peroxide (concentrated peracetic acid) with neurologic sequelae]. AB - The case of accidental, acute oral poisoning with concentrated hydrogen peroxide with peracetic acid components of disinfectant using in food industry (Steridial P) is presented. Neurological sequelae as a result of gas embolisation and supportive and symptomatic treatment are described. PMID- 10466001 TI - [Suicidal self poisoning with disulfiram in a 15-year old boy with resulting encephalopathy]. AB - Acute intentional overdoses of disulfiram (app. 30 g) is reported in 15-years-old boy. The neurological disturbances as nystagmus, coma, seizures and than impairment of memory and perception were dominated in the clinical picture. During MRI examination the areas of different signal from corpus callosum were detected which can be referred to focus of demyelinisation. The patient recovered and was discharged from the hospital after 30 days. Neuropsychological examinations have suggested dysfunction of CNS. The control MRI examination 90 days later, did not reveal any pathological changes in the brain. PMID- 10466002 TI - [Psychiatric disorders during acute tetraethyl lead poisoning in a woman with hyperthyroidism]. AB - We described clinical course of tetraethyl lead poisoning of 21 years old woman with hyperthyroidism. Psychotic symptoms dominated and they disappeared after 4 weeks lasting treatment. Therapy is discussed, including toxicologic, psychiatric and endocrynologic treatment. The basic elements of differential diagnosis in toxic and endocrinologic psychosis are discussed. PMID- 10466003 TI - [The usefulness of Aspargin for supplementing potassium and magnesium in ethanol dependent patients]. AB - Intoxication with many xenobiotics cause serious electrolyte abnormalities which require intravenous supplementation. In chronic and acute but not very severe intoxication an oral supplementation of ions can be sufficient. The aim of the study was to evaluate the usefulness of Aspargin for supplementation of potassium and magnesium in chronic alcoholics suffering from withdrawal syndrome. There were 46 study patients, dependent on ethanol treated at the Detoxification Unit of the Department of Clinical Toxicology in Krak w. A differently intensified symptoms of ethanol withdrawal syndrome were noted according to CIWA-A scale in all the patients. Concentration of potassium and magnesium was monitored through the hospitalisation. Except a basic treatment (fluid supplementation, benzodiazepines) also Aspargin (three times a 2 tablets per day) was administered to each of patient for 7 to 10 days. No respiratory, muscular and gastrointestinal symptoms due to hypopotassemia and hypomagnesemia have been noted while hospitalisation in any patient so it could be suspected, that oral supplementation with Aspargin was sufficient to keep these ions balanced. Administration of Aspargin was beneficial to patient and the treatment outcome. PMID- 10466004 TI - [Incidence and progression of diabetic retinopathy in low income population of Mexico City]. AB - Diabetic retinopathy is one of the leading causes of preventable blindness in working age population. Diabetes mellitus and this microvascular complication affects frequently Mexican population and presents itself in severe clinical forms. There are no incidence studies of diabetic retinopathy in Mexico. The four year incidence and progression of diabetic retinopathy were investigated in low income diabetic patients of Mexico City. In the follow up phase we studied 164 patients, 76.6% of the patients studied at baseline, 63 were men and 101 women. All participants had a complete ophthalmological exam and seven field stereo photographs. All photographs were graded using internationally accepted criteria in the reading center of our institution. The four year incidence of any level of retinopathy was 22.5%. Worsening of retinopathy occurred in 20.6% and the proliferative diabetic retinopathy stage was reached in 4.5%. Incidence of diabetic retinopathy was associated to age at diagnosis of diabetes mellitus of less than 45 years and progression was associated to duration of disease of more than ten years. The four year incidence of macular edema was 8.8%. These data are important to plan strategies for prevention of blindness and the implementation of optimal care of diabetic patients in our country. PMID- 10466005 TI - [Risk markers and high blood pressure. The Cienfuegos global project. Longitudinal study 1992-1994]. AB - OBJECTIVE: To determine the significance of the selected risk markers in the development of high blood pressure in a relatively short period of time. DESIGN: Longitudinal or cohort study of a sample of a population from the municipality of Cienfuegos, as a second measurement of the Cienfuegos Global Project. CONTEXT: Community. Municipality of Cienfuegos; geographic area of the subject-object intervention-investigation of the Cienfuegos Global Project. SUBJECT: 1,369 adults aged 15 years of more (619 men and 750 women; 1,294 alive and 75 dead) were followed from January 1992 to February 1994, which constitutes an alleatory, stratified, equiprobabilistic sample by age and sex. Persons diagnosed as hypertensive in the initial evaluation of Cienfuegos Global Project were excluded, so that the new series for hypertensive patients was constituted by 731 individuals with 125 new patients when the risk markers age, sex, color of skin, family history of high blood pressure, salt consumption, sedentarism, alcoholism and tobaccoism were studied. The association of the risk markers hypercholesterolemia or risky cholesterolemia and obesity was studied with another series formed by a second sample of 417 individuals, this sample introduced 74 new cases of high blood pressure. MAIN OUTCOME: It was intended to estimate the odds ratio for high blood pressure for the different risk markers studied, in those patients exposed or not to them. RESULTS: The odds ratio for each of the risk markers was determined. The odds related to age increased from 1.37, to 1.86 in males and decreased to 0.64 in white individuals. These 3 results were statistically significant as risk markers for high blood pressure. The odds ratio for the remaining risk markers had no statistical significance. CONCLUSION: The age, male sex, and black race, in the comparatively short term- slightly over two years--are the main risk markers that will markedly favor the appearance of high blood pressure. PMID- 10466006 TI - [Bayesian prediction of chloramphenicol blood levels in children with sepsis and malnutrition]. AB - OBJECTIVE: To validate the population pharmacokinetic parameters of chloramphenicol in pediatric patients with sepsis and malnutrition (PPSM) using a bayesian forecasting program. DESIGN: Retrospective evaluation of predictive performance of a bayesian program in PPSM. SETTING: Tertiary care center. PATIENTS: Fifteen MPSP and ten NMPSP that receiving treatment with chloramphenicol. METHODS AND MAIN RESULTS: In the first part of the study, the medical records of 10 MPSP and 10 NMPSP who had received treatment with chloramphenicol were reviewed. The population pharmacokinetic parameter values for each group were estimated using a nonparametric expectation maximization algorithm (NPEM). In the second part, data gathered from five other MPSP receiving chloramphenicol were entered into a bayesian program. Chloramphenicol pharmacokinetic values for each of these five patients were estimated, first using the values of NMPSP as a priori distribution and then repeating the analysis using the MPSP values. The bayesian serum chloramphenicol concentrations predicted for each population model were compared with the actual peaks and troughs. The specific model for MPSP permitted forecasting the peak and trough serum chloramphenicol concentrations with less bias and a better precision compared with the NMPSP population model. CONCLUSIONS: These data indicate that chloramphenicol pharmacokinetics in PPSM can be predicted with minimal bias and good precision using a bayesian forecasting program, allowing a better control of the chloramphenicol serum concentrations. In addition, the limited number of samples required by the bayesian method may represent an important economical benefit for the patient. PMID- 10466007 TI - [Diagnosis of malignant and benign insulinoma. Experience of the National Institute of Nutrition]. AB - INTRODUCTION: Insulinomas are uncommon tumors, their incidence is approximately one case for 1 million population per year. OBJECTIVE: To expose our experience in the diagnosis of these tumors at the Instituto Nacional de la Nutricion Salvador Zubiran. MATERIAL AND METHODS: All patients with histologic diagnosis of insulinoma were reviewed; the period was from 1959 to 1996. The methods used for diagnosis and localization as well as the clinical picture according to the benignity or malignity of the tumor were registered. RESULTS: Eighteen patients were included, four of them with malignant tumors. In two patients islet cell tumors occurred in association with MEN type I, one of them benign and the other malignant. The mean age of patients with benign tumor was 38.2 +/- 13.5 years, in those with malignant tumors it was 51.5 +/- 16.2 years. The median period between symptoms and diagnosis was 34.4 +/- 15 months for benign tumors and 6 +/- 1.1 months for those with malignant tumors (p = 0.02). Patients with increasing weight had benign tumors. The 24 hours fasting test was not done in patients with malignant tumors because of the severity of hypoglycemia. The insulin/glucose ratio in patients with a malignant tumor was 5.2 +/- 4.7, while in patients with a benign tumor it was 1.82 +/- 1.7. The imaging studies showed that three patients with malignant tumors had hepatic metastasis and one had lymph node metastasis. The low accuracy of localization by radiological methods is due to the size of the tumor (> 2 cm). CONCLUSIONS: Malignant insulinomas are more aggressive and the delay of diagnosis is shorter that in cases with benign tumors. Selective arteriography remains the best preoperative localization procedure. PMID- 10466008 TI - [Use of a questionnaire for screening people with undiagnosed diabetes]. AB - OBJECTIVE: To evaluate a screening questionnaire to identify individuals with undiagnosed diabetes. DESIGN AND METHODS: We conducted a community survey to detect people at increased risk for diabetes using a questionnaire proposed by Herman et al. (1995), that incorporates major risk factors as age, obesity, family history of diabetes, sedentary lifestyle and personal history of delivering a macrosomic infant. Blood glucose test was made by means of reagent strip and a reflectance meter system. We used the ADA recommendations and cut points for screening programs, adjusted for fasting and random blood glucose. RESULTS: We included 360 participants older than 20 years of age. A total of 200 subjects (55.5%) were at risk for diabetes according to the questionnaire, of whom 31 (15.5%) had an abnormal glucose test compared to the 4.4% of the low-risk group (p < 0.001). The 1995 Herman et al. Questionnaire had sensitivity of 81.6%, specificity of 47.5, positive predictive value of 15.5 and negative predictive value of 95.6%. The high-risk group was older (44.9 vs. 34.6 y, p < 0.001) and heavier (30.5 vs. 24.4 kg/m2, p < 0.001) than the low-risk group. There were 38/360 (10.5%) abnormal glucose readings, of which 31 (81.5%) had a positive questionnaire (p < 0.01). The mean fasting glucose in the high-risk group was higher (90.6 vs. 84.2 mg/dL, p = 0.015) than in those with low risk by questionnaire, as well as for random blood glucose (116.1 vs. 100 mg/dL, p < 0.01). CONCLUSIONS: The questionnaire proposed by Herman et al. combined with capillary blood glucose testing performance good in mexican population to identify people at high risk for undiagnosed diabetes, and improved the detection rate. PMID- 10466010 TI - [Reflections on the rejections of articles in La Revista de Investigacion Clinica]. PMID- 10466009 TI - [Integrins and integrin-associated molecules: targets for the development of antimetastatic therapies]. AB - Integrins are receptors that mediate cell adhesion and the formation of signaling complex. Changes in the expression of integrins are required during the following steps in the generation of metastases: a) angiogenesis; b) detachment from the primary tumor; c) tumor cell-platelet interaction; d) adhesion to vascular endothelium and e) proliferation. There is a correlation between invasive capability and changes in the expression of some proteins that are clustered in focal adhesion sites, as FAK, CD82, CD9 or CD63. Both, integrin blocking (using antibodies or RGD containing peptides), as well as induced changes in the expression of integrin-associated molecules, are able to inhibit formation of metastases. Discovery and characterization of molecules that regulate the adhesive capability of tumor cells, will lead to development of antimetastasic therapies. In the search of tumor dissemination inhibitors, integrins and some integrin-associated molecules are important pharmacological targets. PMID- 10466012 TI - Hip fracture outcomes and their prevention in Kanawha County, West Virginia. AB - Fracture of the hip is a serious and frequent problem in elderly women, often resulting in a considerable loss of function and an increased mortality long after the initial episode. By reviewing medical records coded for fracture of the proximal femur, we determined the incidence of hip fracture and the in-hospital outcomes for women ages 50 and older in Kanawha County during 1995-96. Additionally, we estimated from existing sources the potentially treatable risk factors for prevention of this malady. Our discharge outcomes showed that ambulation status, discharge to home and the mortality rate were similar to national averages; however, the incidence of hip fracture in our subjects was higher than in the most age-specific groups. Physical inactivity, impaired vision, falls, smoking, alcohol intake and a low bone mineral density were identified in our area as significantly prevalent and potentially treatable risk factors. Although hip-fracture prevention can be approached by the individual health care practitioner, community-based programs would appear to be more practical; however, little evidence currently exists to support this type of large scale intervention. PMID- 10466013 TI - Dyslipidemia evaluated by brachial artery vasoactivity: a case study of eight patients. AB - Current National Cholesterol Education Program (NCEP) guidelines consider a desirable low-density lipoprotein cholesterol level to be < 100 mg/dl for an individual with coronary artery disease (CAD). To assess the affect of these cholesterol levels on endothelial cell functioning, flow-mediated brachial artery vasoactivity was measured non-invasively (7.5 MHz ultrasound) before and during Simvastatin (Zocor) therapy. Vasoactivity was expressed as a percent diameter change from baseline to hyperemic conditions and increased from 2.4 +/- 3.0% at baseline to 13.1 +/- 3.4% (p < .05) and 15.1 +/- 2.7% (p < .01) as cholesterol decreased from 243 +/- 24.7 to 221 +/- 27.3 and 213 +/- 22 mg/dl after three and six months, respectively, during therapy. LDL cholesterol subsequently decreased from 155 +/- 22.1 at baseline to 143 +/- 20.2 and 130 +/- 10.6 after three and six months of treatment, respectively. In conclusion, vasoactivity was found to correlate inversely with cholesterol levels (r = 0.44). This suggests that vasoactivity responds to changes in cholesterol levels and that endothelial function improves by lowering cholesterol levels. PMID- 10466014 TI - Status of women in academic medicine at West Virginia University School of Medicine. AB - The committee on Women in Academic Medicine at West Virginia University conducted a study to compare the status of women in academic medicine at West Virginia University with national averages. We obtained data on gender distributions, ranks, degrees, and administrative titles of full-time women faculty members. Our data showed that 26% of WVUSOM faculty are women. This fell short of the national average within some departments and exceeded the national average in others. Our data also indicated that among women faculty, 15% are full professors (10% nationally), 27% are associate professors (19% nationally), 51% are assistant professors (50% nationally), and 7% are instructors (17% nationally). No clinical departments, and only two basic science departments, are chaired by women. We conclude that the proportion of women faculty and differences in rank distributions at WVUSOM mirror national trends with few exceptions. There is a lack of women in senior positions including full professors, departmental chairs, division chiefs, and academic deans at WVUSOM. PMID- 10466015 TI - Changes in exercise and post-exercise core temperature under different clothing conditions. AB - This study evaluates the effect of different levels of insulation on esophageal (Tes) and rectal (Tre) temperature responses during and following moderate exercise. Seven subjects completed three 18-min bouts of treadmill exercise (75% VO2max, 22 degrees C ambient temperature) followed by 30 min of recovery wearing either: (1) jogging shoes, T-shirt and shorts (athletic clothing); (2) single knit commercial coveralls worn over the athletic clothing (coveralls); or (3) a Canadian Armed Forces nuclear, bacteriological and chemical warfare protective overgarment with hood, worn over the athletic clothing (NBCW overgarment). Tes was similar at the start of exercise for each condition and baseline Tre was approximately 0.4 degree C higher than Tes. The hourly equivalent rate of increase in Tes during the final 5 min of exercise was 1.8 degrees C, 3.0 degrees C and 4.2 degrees C for athletic clothing, coveralls and NBCW overgarment respectively (P < 0.05). End-exercise Tes was significantly different between conditions [37.7 degrees C (SEM 0.1 degree C), 38.2 degrees C (SEM 0.2 degree C and 38.5 degrees C (SEM 0.2 degree C) for athletic clothing, coveralls and NBCW overgarment respectively)] (P < 0.05). No comparable difference in the rate of temperature increase for Tre was demonstrated, except that end-exercise Tre for the NBCW overgarment condition was significantly greater (0.5 degree C) than that for the athletic clothing condition. There was a drop in Tes during the initial minutes of recovery to sustained plateaus which were significantly (P < 0.05) elevated above pre-exercise resting values by 0.6 degree C, 0.8 degree C and 1.0 degree C, for athletic clothing, coveralls, and NBCW overgarment, respectively. Post-exercise Tre decreased very gradually from end-exercise values during the 30 min recovery. Only the NBCW overgarment condition Tre was significantly elevated (0.3 degree C) above the athletic clothing condition (P < 0.05). In conclusion, Tes is far more sensitive in reflecting the heat stress of different levels of insulation during exercise and post-exercise than Tre. Physiological mechanisms are discussed as possible explanations for the differences in response. PMID- 10466016 TI - Impairment of renal sodium excretion in tropical residents--phenomenological analysis. AB - There is evidence of impaired renal sodium excretion in salt-sensitive African Blacks. A decreased rate of renal sodium chloride (NaCl) excretion, low plasma renin activity and a tendency to elevated blood pressure are the hallmarks of salt sensitivity. Recent evidence indicates that increased proximal and distal tubular fluid reabsorption in some tropical residents may explain the impaired sodium excretion in these people. In this study of a cohort population, we speculated that subjects selected from that population might be salt-sensitive. We therefore measured the sodium balance in 10 normotensive male subjects over 10 consecutive days, after they had ingested a normal or a high amount of sodium, as NaCl (salt) in their diet. We quantified their renal sodium excretion rate by phenomenological analysis of their sodium balance data. We also measured plasma renin activity for 7 consecutive days in a separate group of 6 male and 4 female subjects in order to assess the state of their renin/angiotensin system. We selected all our subjects from a cohort population of 269 subjects randomly selected from a community known to have a high prevalence of primary hypertension. Our data on two separate groups of subjects from the same cohort population revealed delayed renal sodium excretion with t1/2 of about 5 days, compared to published data for normal individuals with t1/2 of less than 24 h. Also, plasma renin activity levels were low. Hence, our subjects are salt sensitive. Quantification of their renal impairment is important for various reasons: it heightens one's appreciation of the problem of salt retention in African Blacks who are salt-sensitive and it also underlines the importance of the need for further research into the benefits of dietary salt restriction for reducing cardiovascular mortality in African populations, as has been done in some Western countries. PMID- 10466017 TI - Daily hospital respiratory admissions and winter air mass types, Birmingham, UK. AB - A synoptic climatological approach is used to investigate linkages between air mass types (weather situations), the daily mean particulate matter with a size of 10 microns or less (PM10) concentrations and all respiratory hospital admissions for the Birmingham area, UK. Study results show distinct differential responses of respiratory admission rates to the six winter air mass types identified. Two of the three air masses associated with above average admission rates (continental anticyclonic gloom and continental anticyclonic fine and cold) also favour high PM10 levels. This association is suggestive of a possible linkage between weather, air quality and health. The remaining admissions-sensitive air mass type (cool moist maritime) does not favour high PM10 levels. This is considered to be indicative of a direct weather-health relationship. A sensitising mechanism is proposed to account for the linkages between air mass type, PM10 concentrations and respiratory response. PMID- 10466018 TI - The effects of extra-low-frequency atmospheric pressure oscillations on human mental activity. AB - Slight atmospheric pressure oscillations (APO) in the extra-low-frequency range below 0.1 Hz, which frequently occur naturally, can influence human mental activity. This phenomenon has been observed in experiments with a group of 12 healthy volunteers exposed to experimentally created APO with amplitudes 30-50 Pa in the frequency band 0.011-0.17 Hz. Exposure of the subjects to APO for 15-30 min caused significant changes in attention and short-term memory functions, performance rate, and mental processing flexibility. The character of the response depended on the APO frequency and coherence. Periodic APO promoted purposeful mental activity, accompanied by an increase in breath-holding duration and a slower heart rate. On the other hand, quasi-chaotic APO, similar to the natural perturbations of atmospheric pressure, disrupted mental activity. These observations suggest that APO could be partly responsible for meteorosensitivity in humans. PMID- 10466019 TI - Heat and mass exchange processes between the surface of the human body and ambient air at various altitudes. AB - The rates of convection and evaporation at the interface between the human body and the surrounding air are expressed by the parameters convective heat transfer coefficient hc, in W m-2 degrees C-1 and evaporative heat transfer coefficient h(e), W m-2 hPa-1. These parameters are determined by heat transfer equations, which also depend on the velocity of the airstream around the body, that is still air (free convection) and moving air (forced convection). The altitude dependence of the parameters is represented as an exponential function of the atmospheric pressure p: hc approximately pn and h(e) approximately p1-n, where n is the exponent in the heat transfer equation. The numerical values of n are related to airspeed: n = 0.5 for free convection, n = 0.618 when airspeed is below 2.0 ms-1 and n = 0.805 when airspeed is above 2.0 ms-1. This study considers the coefficients hc and h(e) with respect to the similarity of the two processes, convection and evaporation. A framework to explain the basis of established relationships is proposed. It is shown that the thickness of the boundary layer over the body surface increases with altitude. As a medium of the transfer processes, the boundary layer is assumed to be a layer of still air with fixed insulation which causes a reduction in the intensity of heat and mass flux propagating from the human body surface to its surroundings. The degree of reduction is more significant at a higher altitude because of the greater thickness of the boundary layer there. The rate of convective and evaporative heat losses from the human body surface at various altitudes in otherwise identical conditions depends on the following factors: (1) during convection--the thickness of the boundary layer, plus the decrease in air density, (2) during evaporation (mass transfer)--the thickness of the boundary layer, plus the increase with altitude in the diffusion coefficient of water vapour in the air. The warming rate of the air volume due to convection and evaporation is also considered. Expressions for the calculation of altitude dependences hc (p) and h(e) (p) are suggested. PMID- 10466021 TI - Cumulative subject and contributor indexes and tables of contents for volumes 1 33. PMID- 10466020 TI - Conidia of Alternaria in the atmosphere of the city of Cordoba, Spain in relation to meteorological parameters. AB - In this study, we have analyzed the presence of conidia belonging to different species of the genus Alternaria in the atmosphere of the city of Cordoba, using a Hirst sampler. The results show that spores of this genus are present all year, with a clear seasonal pattern which shows two peaks, one in spring and the other in fall. A total of 26,822 conidia/m3 have been sampled, which implies a daily mean of 74.3 conidia/m3. Statistical analyses comparing the data with meteorological parameters show a positive correlation with maximum, minimum and mean temperatures, and a negative correlation with rain. Nevertheless, meteorological parameters seem to affect the number of conidia differently according to the season of the year. Regression analyses carried out in order to obtain a predictive pattern show that the best fit is between the 7-day running mean of the number of conidia and a week's accumulated mean temperature. PMID- 10466022 TI - Research Society of Alcoholism scientific conference. Santa Barbara, California, USA. June 26-July 1, 1999. Abstracts. PMID- 10466023 TI - 3rd Symposium on Specific Allergy '99. Copenhagen, Denmark, April 15-17, 1999. Abstracts. PMID- 10466024 TI - XII World Congress of the International Society for Artificial Organs and XXVI Congress of the European Society for Artificial Organs. Edinburgh, United Kingdom, 3-6 August 1999. Abstracts. PMID- 10466025 TI - Stereotaxic atlas of the pig brain. AB - A stereotaxic atlas of the pig brain has been established. It consists of 60 frontal and 18 sagittal drawings illustrating Nissl stained sections. A stereotaxic apparatus adapted for the Pig was used to determine the brain coordinates. Radiographic techniques were applied to verify the correct position of the central nervous structures in the apparatus. The zero horizontal plane contains the line joining the recessus preopticus and the posterior commissure (PC) images. These two landmarks were identified by ventriculography. The anterior limit of the PC is used as zero on the stereotaxic atlas. Vertical electrode trajectories implanted at a fixed depth in the brain and at different anterior and lateral levels were used to determine the lateral, vertical and height coordinates in the different planes. Then the brains were perfused and embedded to avoid major displacement of the cerebral structures. Frontal and sagittal serial sections of 100 microns were performed in two different animals. Magnified diagrams of brain sections representing the cellular architecture of the brain structures were produced, some of which are illustrated with photomicrographs. For certain areas, the limits of the nuclei were difficult to determine even when complementary electrophysiological studies were carried out. Each plate is accompanied by a list of abbreviations used to label structures on the plate, together with the terms which they represent. This work should provide a useful anatomical guide for research on the pig brain. PMID- 10466026 TI - XIX International Conference on Yeast Genetics and Molecular Biology. Rimini, Italy, 25-30 May 1999. Abstracts. PMID- 10466027 TI - Federation of the European Societies for Surgery of the Hand 6th Congress Instructional Course. Fractures of the metacarpals and phalanges in adults and children. Bonn, Germany, 26-29 May 1999. Abstracts. PMID- 10466028 TI - 9th International Conference on Biological Inorganic Chemistry. Minneapolis, Minnesota, USA. July 11-16, 1999. Abstracts. PMID- 10466029 TI - 53rd Annual meeting of the Canadian Society of Otolaryngology--Head and Neck Surgery. Halifax, Nova Scotia, Canada. June 20-23, 1999. Abstracts. PMID- 10466030 TI - European Society for Therapeutic Radiology and Oncology annual brachytherapy meeting. Utrecht, The Netherlands, 10-12 May 1999. Abstracts. PMID- 10466031 TI - Joint meeting of the 50th annual meeting of the German Society of Neurosurgery with the Swiss Society of Neurosurgery. Munich, Germany, 5-9 June 1999. Abstracts. PMID- 10466032 TI - The structure of major birch pollen allergens--epitopes, reactivity and cross reactivity. PMID- 10466033 TI - Can the natural course of allergy and asthma be changed by allergen vaccinations? PMID- 10466034 TI - Allergen-specific immunotherapy. Present state and directions for the future. PMID- 10466035 TI - Mechanisms of mucosal inflammation in the nose and lungs. PMID- 10466036 TI - Pharmacological modulation of the allergic response: different modes of action. PMID- 10466037 TI - Inhaled corticosteroids versus anti-leukotrienes: a literature review on the clinical effects. PMID- 10466038 TI - What is the optimal way of treating these patients today and in the future? PMID- 10466039 TI - Resistance to Fas (APO-1/CD95)-mediated apoptosis and expression of Fas ligand in esophageal cancer: the Fas counterattack. AB - The mechanisms by which esophageal tumors escape immunologic recognition and clearance are only partly understood at the molecular level. Esophageal cancers have been shown to evade host recognition by down-regulation of antigen presentation and production of immunosuppressive factors. Recently, two independent reports have shown that esophageal tumor cells abundantly express Fas ligand (FasL) in vivo. As the triggering agonist for Fas receptor (Fas or APO 1/CD95)-mediated apoptosis of lymphocytes, FasL normally plays immune down regulatory roles, including activation-induced cell death of T and B cells, as well as maintaining immune privilege in certain organs. Fas ligand expressed by esophageal cell lines has been shown to induce apoptosis of cocultured Fas sensitive lymphoid cells in vitro. FasL expression by esophageal carcinomas in vivo has been associated with significantly reduced tumor-infiltrating lymphocytes (TILs) in FasL-positive tumor nests, concomitant with significantly increased TIL apoptosis in these nests. These studies support a 'Fas counterattack' mechanism of immune escape in esophageal cancer. By expressing functional Fas ligand, esophageal cancer cells can deplete antitumor lymphocytes by inducing apoptosis. To express functional FasL, esophageal carcinomas also acquire molecular mechanisms to resist autocrine Fas-mediated apoptosis of tumor cells. PMID- 10466040 TI - Fas ligand and Fas receptor are coexpressed in normal human esophageal epithelium: a potential mechanism of apoptotic epithelial turnover. AB - Fas (CD95/Apo-1) receptor (FasR) is a cell-surface receptor that mediates apoptotic cell death upon triggering by Fas ligand (FasL). We sought to determine whether normal human esophageal epithelial cells express FasL and/or FasR and whether their localization is consistent with a role in the turnover of normal esophageal epithelium. Normal esophageal epithelium was immunohistochemically positive for FasL in upper prickle cell layers and in mature squamous cells, but the proliferative basal layer was negative. FasL mRNA was detected in the same epithelial cell layers by in situ hybridization. Co-localization of FasL mRNA and protein therefore confirmed that FasL expression is induced in esophageal epithelial cells as they reach terminal differentiation. FasR was immunohistochemically detected throughout the esophageal epithelium. Positive TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling) staining confirmed cell death of the FasL and FasR coexpressing mature epithelial cells. CD45-positive immunocytes were notably absent from FasL expressing upper epithelial layers. The findings are consistent with a contributory role for Fas-mediated autocrine suicide or paracrine fratricide in the apoptotic turnover of normal esophageal epithelium. PMID- 10466041 TI - Induction of esophageal carcinogenesis by diethylnitrosamine and assessment of the promoting effect of ethanol and N-nitrosonornicotine: experimental model in mice. AB - It is known that some nitrosamines preferably affect particular organs because of their organospecificity. Diethylnitrosamine (DEN) is one of the most powerful nitrosamines for experimentally inducing esophagus cancer. The present study aimed to evaluate the rate and type of epithelial lesions induced by DEN in mice. We also assessed the role of alcohol and N-nitrosonornicotine (NNN) as promoters of this carcinogenesis. A total of 208 female mice (Mus musculus) were allocated to five experimental groups: group 1, water only (controls); group 2, DEN + water; group 3, DEN + NNN; group 4, DEN + 6% alcohol solution; group 5, DEN + NNN + 6% alcohol solution. Animals in groups 2, 3, 4 and 5 received DEN (0.04 ml/l) three times per week, and during the following 4 days they received the other solutions. NNN was provided at a final concentration of 30 mg/l. The overall experimental period was 180 days. At the end of this time, the animals were killed and their esophagus was dissected for macro- and microscopic analysis. There was no significant difference in relation to the size of the esophagus and to the average DEN intake by the animals (p > 0.05). A statistically significant difference (p < 0.0001) was observed between controls and all other experimental groups. There was no significant difference among experimental groups treated with carcinogens (p > 0.05). The average incidence of cancer was 85.4%. The experimental model used in the present study is a very potent indicator of esophagus cancer. Owing to the high incidence for cancer observed in the present study, it was not possible to assess the effect of alcohol and NNN as inducers for the development of esophageal cancer. PMID- 10466042 TI - Influence of surgically induced gastric and gastroduodenal content reflux on esophageal carcinogenesis--experimental model in Wistar female rats. AB - Studies in human beings and animals have shown that esophageal exposure to duodenal and gastric contents may be important for the development of Barrett's esophagus and its complications, including adenocarcinoma and epidermoid carcinoma. Diethylnitrosamine (DEN) is a carcinogen that stimulates the development of epidermoid carcinoma in the esophagus of mice. The aim of this study was to evaluate the effect of gastroduodenal and gastric content reflux on induction of esophageal carcinogenesis. Gastroesophageal reflux (GER) and gastroduodenoesophageal reflux (GDER) were produced by cardioplasty and esophagoduodenostomy. The chosen carcinogen was DEN, diluted in drinking water, given 3 days a week for 20 consecutive weeks. One hundred Wistar female rats were divided into six groups, as follows: group 1 (18 rats), cardioplasty without DEN; group 2 (18 rats), cardioplasty with DEN; group 3 (10 rats), only water; group 4 (17 rats), cardioplasty with DEN; group 5 (17 rats), esophagoduodenostomy with DEN; group 6 (20 rats), only DEN. GER in isolation induced papillomatosis or ulceration in 22.2% of rats and, when associated with DEN, induced papillomatosis in 61.1% of rats. GDER in isolation induced marked esophagitis in 61.1% of rats, Barrett's esophagus in 16.7% and esophageal adenocarcinoma in 16.7%; when associated with DEN, 23.5% of rats presented marked esophagitis, papillomatosis or ulceration, whereas 76.5% had esophageal carcinoma, with 70.6% epidermoid carcinoma and 5.9% adenocarcinoma. Rats treated with water alone did not show histologic abnormalities of the esophageal mucosa. Rats treated with DEN alone developed papillomas in 50.0% of the cases and remained histologically unchanged in 50.0%. There was no development of low- or high-grade dysplasia in any group. The conclusions are that (1) GDER is significantly more deleterious to esophageal mucosa than GER; (2) in this study, GER did not present carcinogenic potential in relation to the esophagus; (3) GDER in isolation is an esophageal carcinogen, producing Barrett's esophagus and esophageal adenocarcinoma; (4) esophageal oncogenesis caused by GDER is potentiated by DEN, inducing esophageal epidermoid carcinoma; (5) in this study, DEN in isolation did not generate tumors in the esophagus of rats. PMID- 10466043 TI - p53, p21(Waf1/Cip1) and cyclin D1 protein expression and prognosis in esophageal cancer. AB - Recently, various cell cycle regulators have been investigated as biological markers of malignant potential. These regulators might influence the survival rate and the effect of adjuvant therapies. In this study, we analyzed p53, p21(Waf1/Cip1) and cyclin D1 expression in 64 esophageal cancer patients and the relationship between clinicopathologic parameters and patient survival. The positive expression rate was 48.4%, 42.2% and 43.8% in the p53, p21 and cyclin D1 groups respectively. Multivariant analysis revealed that tumor depth, chemotherapy, p53, p21 and cyclin D1 expression showed significant values. p53- and cyclin D1-negative patients had a worse prognosis. p21-positive patients had a better prognosis. In stage 0, I and II patients, there was a significant difference between p53-positive and -negative, p21-positive and -negative, and cyclin D1-positive and -negative groups. In stage III and IV patients, there was no significant difference between any two groups. However, a significant difference was seen in the p21 group: among patients who received adjuvant chemotherapy, the p21-positive group had a 5-year survival rate of 50% compared with 13.4% in the p21-negative group (not significant). PMID- 10466044 TI - The prognostic significance of cell cycle markers in esophageal cancer after neoadjuvant chemotherapy. AB - Proliferating cell nuclear antigen (PCNA), p27 and cyclin A were analyzed by immunohistochemistry in 89 patients (untreated control n = 40, neoadjuvant chemotherapy n = 49) with esophageal cancer invading the submucosal lesion. The mitotic index (MI) was calculated as the percentage of mitotic cells. In control subjects, the mean PCNA, p27, cyclin A and MI were, respectively, 60.4%, 18.0%, 19.9% and 1.7%; in the chemotherapy group, these values were 46.8%, 15.1%, 18.0% and 1.2% respectively. Neoadjuvant chemotherapy decreased PCNA and the MI significantly. As prognostic indicators, PCNA and the MI were significant in control subjects and p27 and cyclin A were significant in the chemotherapy group. Using multivariate analysis, p27 was a prognostic factor in both groups and cyclin A was prognostic only in the chemotherapy group. Although PCNA and the MI were useful growth and prognostic markers in untreated control subjects, their significance was lost after neoadjuvant chemotherapy. p27 and cyclin A were determined to be significant markers in the neoadjuvant chemotherapy group, especially p27, which was independent in both groups. PMID- 10466045 TI - p53 Protein accumulation as a prognostic marker of preoperative radiotherapy and/or chemotherapy in advanced squamous cell esophageal carcinoma--preliminary report. AB - The correlation between immunohistochemical detection (IH) of p53 protein and tumor response to preoperative chemotherapy and/or radiotherapy in advanced esophageal squamous cell carcinoma was evaluated. Fifty-six patients with advanced esophageal squamous cell carcinoma were included in the study. All patients were staged and diagnosed microscopically before treatment. Patients were divided into three groups: 17 patients treated with chemotherapy and radiotherapy preoperatively (group I) (cisplatin and 5-fluorouracil, cobalt-60 therapy; total dose 3000 Gy); 19 patients treated with chemotherapy only (group II); and 20 patients who did not receive preoperative therapy (group III). The response of the tumor tissue to preoperative treatment was evaluated macroscopically and microscopically in operated specimens according to the classification: CR, complete response; PR1, major partial response with regression of at least 50% of initial tumor mass; PR2, minor partial response with regression of less than 50% of initial tumor mass. In all 56 patients immunohistochemistry was used to detect anti-p53 antibody (Dako, DO-7) in normal mucosa and cancer tissue. The response of the tumor was similar in both group I and group II. p53 protein was not expressed in the normal esophageal mucosa. A high level of p53 in operated specimens was associated with unfavorable tumor response to preoperative treatment. Therefore, immunohistochemical detection of p53 protein can be considered to predict the outcome of preoperative therapy. PMID- 10466046 TI - Microsatellite instability in double cancers of the esophagus and head and neck. AB - It is generally accepted that patients with squamous cancers of the esophagus are known to have a high risk of concomitant head and neck cancer. However, there have been only a few reports describing microsatellite instability (MSI) in patients with both esophageal squamous cell carcinoma and head and neck cancers. To evaluate the role of genetic instability in carcinogenesis in such patients, we analyzed six microsatellite loci in 21 tumors from 10 patients who had developed primary cancers of both the esophagus and the head and neck. MSI was detected in 6 out of 10 patients. In five patients with double cancer, MSI was observed at the same microsatellite loci in both the esophageal and the head and neck tumors obtained from the same individuals. These data suggest that such patients may have the same underlying defect in the mismatch repair system, providing insight into possible mechanisms for field carcinogenesis. PMID- 10466047 TI - Two-field radical lymphadenectomy in the treatment of esophageal carcinoma. AB - This paper retrospectively compares post-operative complications, mortality and long-term survival of patients with esophageal carcinoma who were treated with standard esophagectomy or with extended two-field lymph node clearance. Fifty seven patients with resectable esophageal carcinoma were included in the study. Twenty-eight patients were submitted to a radical two-field esophagectomy and lymphadenectomy, while the remaining 29 were submitted to a standard, more conservative, esophagectomy performed mostly through a transhiatal route. The two groups of patients were similar in all clinical, laboratory and pathologic features. There was a significant lower anastomotic leakage rate in the group of patients submitted to a radical lymph node resection; post-operative respiratory complication rate and mortality were similar in both groups. The overall 5-year survival was 20%. When lymph node resection was performed, the 5-year survival rate rose to 36%; it was 44% when nodal involvement was negative and 19% for N1 patients; when standard esophagectomy was the procedure, these figures were 9% (p < 0.05), and 6% respectively. PMID- 10466048 TI - Neoadjuvant chemotherapy before surgery for resectable carcinoma of the lower esophagus. AB - Neoadjuvant chemotherapy before surgery has been proposed to improve the outcome in patients with early lower esophageal cancer. To evaluate its effectiveness, we performed a systematic retrospective analysis of consecutive patients treated at the Ottawa Regional Cancer Center with prospective inclusion criteria. Between 1988 and 1992 patients were treated with surgery alone. From 1992 until 1997, patients were uniformly treated with neoadjuvant chemotherapy consisting of cisplatin and 5-fluorouracil. Surgical resection was then performed. Nineteen patients received neoadjuvant chemotherapy and 15 received surgery alone. Although the two arms of the study were balanced for age and sex, there were more patients in the neoadjuvant arm with squamous histology, weight loss and regional nodes at diagnosis. In the neoadjuvant arm, two patients did not have surgery because of progression or toxicity. However, complete resection rates were similar. There was no difference in overall survival or disease-free survival between the two arms (p > 0.4). Multivariate analysis revealed that only the nodal status at diagnosis was predictive of outcome. Neoadjuvant chemotherapy with this regimen does not result in improved survival over surgery alone. PMID- 10466049 TI - Esophagogastric anastomotic wound healing in rats. AB - Esophagectomy with esophagogastric anastomosis is commonly complicated by anastomotic dehiscence. Although this is a major problem in clinical esophageal surgery, laboratory investigation of esophagogastric anastomotic wound healing has been hampered by the lack of a practical rodent model. Problems with aspiration pneumonia and anastomotic strictures hindered our previous studies in the rat. Other researchers have turned to large animal experiments, or used various upper gastrointestinal pseudoanastomotic techniques in rodents. None of these approaches has proved satisfactory. We developed a technique of side-to side esophagogastric anastomosis in the rat, and then studied normal esophagogastric anastomotic healing in this model. Anastomoses were performed in 24 rats. Anastomotic breaking strength and hydroxyproline concentration were measured 1 and 2 weeks after surgery. Anastomotic breaking strength was 3.78 +/- 1.18 N at 1 week and 4.83 +/- 0.91 N after 2 weeks (p < 0.03). Anastomotic tissue hydroxyproline concentration was 370.6 +/- 31.2 nmol/mg at 1 week and 462.1 +/- 69.7 nmol/mg after 2 weeks (p < 0.001). Many of the problems encountered in esophagogastric anastomotic studies in the rat have been overcome using this new model. PMID- 10466050 TI - Poor prognosis in esophageal cancer patients with post-operative complications: surgical oncotaxis. PMID- 10466051 TI - Successful management of post-esophagectomy chylothorax/chyloperitoneum by etilefrine. AB - Post-operative chylothorax and chyloperitoneum resulting from a thoracic duct injury are possible complications of esophagectomy for carcinoma. Management of such conditions includes conservative methods (pleural drainage, chyle flow reduction and supportive nutrition) and surgery. Etilefrine, a sympathomimetic drug used in the management of postural hypotension, also causes smooth muscle contraction of the thoracic duct. We successfully treated three patients with post-esophagectomy chylothorax and chyloperitoneum using intravenous etilefrine. Etilefrine infusion allowed chyle flow resolution in a few days without any side effects. As result of this experience, we believe that etilefrine deserves to be considered as an alternative therapy in the management of post-esophagectomy chylothorax/chyloperitoneum. PMID- 10466052 TI - Tumor seeding of the jejunostomy site after transhiatal esophagectomy for esophageal carcinoma. AB - A 65-year-old male patient with squamous cell carcinoma of the esophagus had a transhiatal esophagectomy after a prophylactic tube jejunostomy. The tube was removed 3 weeks after surgery. Ten months later, a painless 2-cm abdominal mass was noted at the previous jejunostomy site. Subsequent segmental resection of the jejunum disclosed metastatic squamous cell carcinoma of the esophagus. It is possible that tumor seeding may develop at the jejunostomy site after transhiatal esophagectomy for esophageal carcinoma. PMID- 10466053 TI - Rational drug design using trypanothione reductase as a target for anti trypanosomal and anti-leishmanial drug leads. AB - The parasite enzyme trypanothione reductase has been used as a target for rational drug design against trypanosomiasis and leishmaniasis in a number of laboratories. In this article the biochemical basis for its selection as a target is reviewed. The relevant structural aspects of the target are then compared with the homologous structure found in the mammalian hosts to indicate the molecular basis by which selective toxicity is likely to be achieved. An overview of known classes of inhibitors is provided, preparatory to a detailed coverage of approaches that have been taken to obtaining strong, selective inhibitors and the steps taken in the process of the initial discovery of tricyclic structures by interactive molecular graphics ligand design are outlined. Recent quantitative docking approaches which have been applied to this system are also described. Finally, the biological data of the activity against the various parasitic forms in vitro and in vivo are summarised. PMID- 10466054 TI - Inhibition of dihydrofolate reductases from Toxoplasma gondii, Pneumocystis carinii, and rat liver by rotationally restricted analogues of pyrimethamine and metoprine. AB - Twenty-one conformationally restricted tricyclic pyrimethamine and metoprine analogues with one or two chlorine atoms, or other substituents, at different positions of the phenyl ring were tested for potency and species selectivity against dihydrofolate reductase (DHFR) from Toxoplasma gondii, Pneumocystis carinii, and rat liver. Heterocyclic systems studied included indeno[2,1 d]pyrimidines, benzo[f]quinazolines, and benzo[3,4]cyclohepta[1,2-d]pyrimidines. All but one of the analogues were more potent against T. gondii and rat liver DHFR than against P. carinii DHFR, and those with a one-carbon (CH2) bridge were generally less potent than those with a two-carbon (CH2CH2, CH=CH) or three carbon (CH2CH2CH2) bridge. Although a number of compounds with a two- and three carbon bridge were more potent than pyrimethamine against P. carinii DHFR, and especially T. gondii DHFR, none of them were selective for the P. carinii versus the mammalian enzyme, and only those with a one-carbon bridge showed selectivity approaching that of pyrimethamine for the T. gondii enzyme. Computer-simulated docking into the active site pocket of P. carinii and human DHFR suggested that, as a group, the rotationally restricted tricyclic structures are at a disadvantage relative to pyrimethamine and metoprine, in that torsional relief of unfavorable steric interactions between the chlorine atoms and two critical serine and threonine residues in the active site is prevented by the bridge. PMID- 10466055 TI - Synthesis and biological evaluation of carbocyclic analogues of lipid X: new nonpolar antagonists of LPS induced TNF production. AB - We have synthesised a number of analogues of lipid X, a precursor in the biosynthesis of LPS, some of which exhibit marked antagonism of LPS induced TNF production in vivo. These compounds provide new non-polar leads in the search for a therapy for endotoxic shock. PMID- 10466056 TI - A comparison of backpropagation and generalized-regression genetic-neural network models. AB - The results of the backpropagation (BP) and generalized-regression genetic-neural (GRGN) network were compared using a series of nonpeptide arginine vasopressin VI antagonists. It was shown that both approaches are equivalent with respect to the recognition process while the BP network is superior over GRGN if the sample sizes are lowered by cross-validation. PMID- 10466057 TI - Studies of molecular pharmacophore/receptor models for GABAA/benzodiazepine receptor subtypes: binding affinities of substituted beta-carbolines at recombinant alpha x beta 3 gamma 2 subtypes and quantitative structure-activity relationship studies via a comparative molecular field analysis. AB - Binding affinities of a series of 44 beta-carbolines with various substituents at the 3-, 4-, 6- and 7-positions are reported at 5 distinct recombinant GABAA/benzodiazepine receptor (BzR) subtypes [alpha x beta 3 gamma 2 (x = 1-3, 5, 6)]. Many of these ligands displayed better selectivity for the alpha 1 containing GABAA isoform. The most selective BCCT 2 and SPH 195 (17) displayed potent affinity (Ki = 0.72 and 7.2 nM for the alpha 1 beta 3 gamma 2 receptor subtype, respectively) and an overall selectivity of 20 and 23 fold, respectively, for the alpha 1 beta 3 gamma 2 receptor subtype. These are the most selective ligands in vitro for the alpha 1 containing GABAA/Bz receptor isoform reported to date to our knowledge. QSAR studies of these ligands for each receptor subtype have been carried out via a Comparative Molecular Field Analysis (CoMFA) and an included volume analysis. Geometries and charge distributions of these ligands have been optimized using ab initio methods (J. Med. Chem., 1992, 35, 4001-4010). Active conformations of flexible 3-alkoxylated beta-carbolines have been examined via a CoMFA approach. QSAR studies via CoMFA support the previous hypothesis that beta-carbolines with different intrinsic activities may follow an alternative alignment rule when they bind into the pharmacophore/receptor site of the BzR. Examination of binding affinities of beta carbolines by this modeling strategy has established some of the differences, in particular, topologic differences between the lipophilic pockets in the alpha 1 beta 3 gamma 2, alpha 2 beta 3 gamma 2, alpha 3 beta 3 gamma 2, alpha 5 beta 3 gamma 2 and alpha 6 beta 3 gamma 2 subtypes as well as some of the similarities among the pharmacophore/receptor models of these five distinct GABAA/Bz receptor subtypes. PMID- 10466058 TI - Studies of molecular pharmacophore/receptor models for GABAA/BzR subtypes: binding affinities of symmetrically substituted pyrazolo[4,3-c]quinolin-3-ones at recombinant alpha x beta 3 gamma 2 subtypes and quantitative structure-activity relationship studies via a comparative molecular field analysis. AB - A series of symmetrically substituted pyrazoloquinolinones was synthesized to probe the BzR binding site of different GABAA/Bz receptor subtypes. The affinities of the ligands for different BzR subtypes have been determined by radioligand binding assays on 5 distinct recombinant GABAA receptor isoforms [alpha x beta 3 gamma 2 (x = 1,2,3,5, or 6)]. Most of the ligands synthesized exhibited potent biological activity in vitro. Among them, 3 ligands exhibited enhanced affinity for the alpha 2 beta 3 gamma 2 subtype in comparison to the other subtypes, six ligands demonstrated higher affinity for the alpha 3 beta 3 gamma 2 subtype, while 2 ligands showed some enhanced affinity for the alpha 5 beta 3 gamma 2 subtype. The remainder of the ligands exhibited relatively higher affinities at the alpha 1 containing subtype. To map out the steric and electronic differences between the benzodiazepine binding subtypes, a QSAR analysis by the method of Comparative Molecular Field Analysis (CoMFA) of each receptor subtypes was carried out. PMID- 10466059 TI - [Immunohistochemical study of p53 and Ki-67 antigen expression in bladder carcinoma]. AB - We examined the relationship between the expression of mutant p53 and Ki-67 antigens in urinary bladder transitional cell carcinoma and the pathological and clinical findings. Tissues were obtained from 28 patients with bladder carcinoma who underwent total or partial cystectomy. An ABC immunostaining method and two primary antibodies (DO-7 and MIB-1 antibodies) were used. The percentages of p53 and Ki-67 antigen-positive cells to the total number of cells were regarded as the p53 and Ki-67 labeling indices (LI) respectively. There were no statistically significant correlations between p53 LI and the histological grade or stage, although p53 LI increased slightly in the high grade and high stage group. There was a statistically significant correlation between Ki-67 LI and the histological grade and stage (p < 0.05). The correlation between p53 LI and Ki-67 LI was linear. Some cases had a p53 LI below the mean even though the Ki-67 LI was higher. The clinical course was characteristic of superficial bladder carcinoma initially, but progressed to invasive bladder carcinoma over the next several years. These results suggest that even cases initially diagnosed as superficial bladder carcinoma with a low p53 LI may progress to invasive bladder carcinoma in subsequent years. Therefore, it is important that the patient be followed-up. PMID- 10466060 TI - Prostate specific antigen density for discriminating prostate cancer from benign prostatic hyperplasia in the gray zone of prostate-specific antigen. AB - Serum prostate specific antigen (PSA) is currently the best blood marker for prostate cancer. However, low specificity for detection of prostate cancer, especially in the gray zone of PSA, is a problem. We evaluated the clinical significance of PSA density (PSAD) in gray zone PSA cases with conversion of serum PSA to a Stanford reference value. In a series of histologically confirmed 63 benign prostatic hyperplasia (BPH) patients and 234 prostate cancer patients, 36 BPH patients and 25 prostate cancer patients had gray zone PSA levels. Serum PSA was measured with the Markit-F or Markit-M PA assay. All data were converted to Stanford reference values. We used transabdominal ultrasound to determine prostate volume. PSAD was determined as the serum PSA/prostate volume ratio. The mean PSA values for BPH and prostate cancer were 6.42 +/- 1.80 and 7.80 +/- 2.15 ng/ml (p = 0.0116), respectively, and prostate volume was 33.4 +/- 14.1 ml and 17.1 +/- 8.2 ml, respectively (p < 0.0001). The mean PSAD for prostate cancer was 0.572 +/- 0.363 while that for BPH was 0.218 +/- 0.085 (p = 0.0001). Cut-off values with sensitivity > 90% were 0.218 for PSAD and 30 ml for prostate volume. At these cut-off values, specificity reached 56% for each marker. In discriminating prostate cancer from BPH in the gray zone of PSA, PSAD demonstrated better performance than PSA. PMID- 10466061 TI - [A case of advanced retroperitoneal dedifferentiated liposarcoma treated effectively with high-dose ifosfamide]. AB - We report a case of retroperitoneal dedifferentiated liposarcoma treated effectively with high-dose ifosfamide. A 59-year-old man received tumorectomy and right nephrectomy for the retroperitoneal liposarcoma. Twenty-two months after the operation, the liver metastasis was resected incompletely. Three months later, right pleural, retroperitoneal and peritoneal metastases appeared. With 6 cycles of high-dose ifosfamide therapy these tumors were reduced partially. Ten cycles of this chemotherapy were administered. These tumors showed regrowth at 14 months after administration of high dose ifosfamide. The combination chemotherapy of ifosfamide and doxorubicine or etoposide was not effective after regrowth of the tumors. PMID- 10466062 TI - [Simultaneous and metachronous pelvic bone metastases from renal cell carcinoma necessitating hemipelvectomy: report of two cases]. AB - Two patients, one with and one without a history of nephrectomy for renal cell carcinoma, presented with lower abdominal pain. One of them, a 49-year-old man, had tumors in the right kidney and the right ischiadic bone. He underwent nephrectomy and pathologic findings showed renal cell carcinoma (tubular type, granular cell subtype, INF-gamma, G2). The other patient, a 33-year-old man with a previous history of left nephrectomy for renal cell carcinoma (tubular type, granular cell subtype, INF-alpha, G2), was found to have a large tumor in the pelvis, extending from the pubic and iliac bones to the hip joint. Both patients underwent embolization of the hypervascular mass using a vascular coil followed by hemipelvectomy under general anesthesia. The pathology reports confirmed bone metastases from renal cell carcinoma. Both patients survived surgery and their postoperative courses were uneventful without urinary or bowel incontinence. However, impaired potency was noted in the latter case. Immunotherapy with INF alpha was resumed immediately after surgery. PMID- 10466063 TI - [Collecting duct carcinoma (Bellini duct carcinoma) of the kidney with tumor extension into the inferior vena cava]. AB - We report a case of collecting duct carcinoma (Bellini duct carcinoma) of the left kidney accompanied with a tumor thrombus in the inferior vena cava and the lymph node metastasis. A 69-year-old male presented with gross hematuria and left flank dullness. Computed tomography revealed an isodensity tumor in the left kidney with tumor extension into the inferior vena cava and the regional lymph node swelling. The T1-weighted magnetic resonance image displayed a slightly heterogeneous low-intensity-mass. Renal angiography revealed a hypervascular tumor. We performed left radical nephrectomy with tumor thrombectomy and regional lymphadenectomy. Histopathological examination revealed a collecting duct carcinoma (pT3bN1M0V2a). Seven months after surgery, multiple metastates in bone and liver developed. Then we performed systemic chemotherapy consisting of methotrexate and cisplatin. However, the patient died from the carcinoma 10 months postoperatively. PMID- 10466064 TI - [Emphysematous pyelonephritis complicated with diabetes mellitus: a case report]. AB - A 46-year-old woman with diabetes mellitus was admitted to our hospital due to lower right abdominal pain. Urinalysis did not show marked pyuria. Abdominal computed tomography showed an abnormal gas shadow in the right renal parenchyma. A diagnosis was made of right emphysematous pyelonephritis. Despite aggressive supportive therapy, the patient's condition worsened. Therefore, right nephrectomy was performed. The next day her general condition was markedly improved. We reviewed 122 cases of emphysematous pyelonephritis including our case in the Japanese literature, and discussed its etiology, symptomatology, choice of treatment and prognosis. PMID- 10466065 TI - [Laparoscopic nephropexy for symptomatic nephroptosis: a case report]. AB - We present our clinical findings of laparoscopic nephropexy performed on a 32 year-old woman with symptomatic nephroptosis. Supine and erect excretory urography (DIP) revealed right renal descent of 2.5 vertebral bodies with hydronephrosis. Laparoscopic transperitoneal nephropexy was performed using nonabsorbable sutures, polyglactin net, hernial staplers and tissue adhesive. An upright DIP 1 month postoperatively revealed renal descent of only 1 vertebral body and no hydronephrosis. One year postoperatively, the patient is asymptomatic. A laparoscopic procedure, which is safe, effective and causes minimal morbidity, represents an excellent approach for repair of symptomatic nephroptosis. PMID- 10466066 TI - [Transitional cell carcinoma in the ureter showing inverted proliferation: a case report]. AB - A case of transitional cell carcinoma in the ureter showing inverted proliferation is reported. A tumor in the middle third of the right ureter was found in a 67-year-old male complaining of gross hematuria. Since positive findings of malignancy were obtained in a washing examination of urine cytology, right nephroureterectomy was performed. The gross specimen consisted of a polypoid and pedunculated 22 x 8 mm tumor which showed a smooth surface as in normal ureteral mucosa. Histopathologically, the tumor was lined with normal transitional epithelium but filled with transitional cell carcinoma, grade 1. Diagnosis and treatment of ureteral tumors showing inverted proliferation are discussed. PMID- 10466067 TI - [Combined treatment with CDDP and radiation effective against neuroendocrine carcinoma of the urinary bladder: a case report]. AB - A case of neuroendocrine carcinoma (small cell carcinoma) of the urinary bladder is presented. A 76-year-old man complaining of dysuria visited our clinic on October 31, 1997. On physical examination, a huge mass was palpable in the lower abdomen. Abdominal and pelvic computed tomographic (CT) scan revealed a huge mass, 6.7 x 6.0 cm in size, with extravesical extension in the anterior wall of the urinary bladder and no metastatic lesions. Percutaneous biopsy of the tumor revealed undifferentiated neuroendocrine carcinoma. The value of serum neuron specific enolase (NSE) was 220 ng/ml (normal range: 0-10 ng/ml). Twenty days after the first CT scan, the tumor had grown to be 12.5 x 11.0 cm in size. He was treated with combination therapy of systemic cisplatin and external pelvic radiation and then achieved complete remission on CT scan and biopsy. The value of serum NSE was normalized. Four months later, abdominal CT scan revealed a huge metastastic lesion in the paraaortic and parahepatic regions, but, no local recurrence in the bladder. The value of serum NSE was 240 ng/ml. He was treated with 4 cycles of systemic combination therapy of cisplatin and etoposide. He achieved partial remission (regression rate: 77%) on CT scan after completion of the first 2 cycles, but the tumor showed rapid re-growth and he died of cancer 1 month later despite another 2 cycles. These combination therapies were effective against neuroendocrine carcinoma of the urinary bladder, although, the duration of the effect was short. PMID- 10466068 TI - [A case of interstitial pneumonitis caused by intravesical bacillus Calmette Guerin instillation]. AB - A 61-year-old man was referred to our hospital due to positive urine cytology. He underwent multiple cold punch biopsies of the bladder and the histopathological finding was transitional cell carcinoma (TCC), carcinoma in situ (CIS), grade 3. He was treated with 121.5 mg of bacillus Calmette-Guerin (BCG) (Connaught strain) suspended in 50 ml of saline instilled into the bladder at weekly intervals. After the third instillation he developed a fever up to 39 degrees C, pain on urination and an elevation of liver enzymes. Antituberculous drugs were administered and he was re-admitted for further evaluation. The chest radiograph showed diffuse extensive bilateral lung densities. His chest computed tomographic (CT) scan showed bilateral interstitial pneumonitis. All cultures from his blood, urine, sputum, and bronchoalveolar lavage remained negative for mycobacteria. He was diagnosed as having a hypersensitivity reaction of bilateral lung after immunotherapy with BCG. Pulse steroid therapy was done. The chest radiograph, findings improved and he was clinically asymptomatic after steroid therapy. PMID- 10466069 TI - [Penile carcinoma with humoral hypercalcemia of malignancy (HHM): a case report]. AB - A 57-year-old male, with 1.5 cm tumor in his prepuce, was admitted to our institute in Feb. 1990. Circumcision and inguinal lymph node dissection was performed under the diagnosis of T1 disease of penile carcinoma. Pathological evaluation revealed well-differentiated squamous cell carcinoma (SCC). In April 1996, the patient revealed recurrence of the disease in the right inguinal lymph nodes and the lower abdomen, that was diagnosed to be poorly differentiated SCC. Laboratory findings showed elevation of serum calcium, parathyroid hormone related protein (PTHrP) levels. Immunohistochemical staining confirmed production of PTHrP in the tumor tissue. This is the first case report of penile carcinoma that caused humoral hypercalcemia of malignancy in Japan. PMID- 10466070 TI - [Sertoli cell tumor of the testis: a case report]. AB - A case of Sertoli cell tumor of the testicle is reported. A 33-year-old man visited the Chiba University Hospital with the chief complaint of a painless right testicular swelling on May 1990. The right testis was hard and swollen on palpation. Gynecomastia was not present. Serum levels of tumor markers and hormones including alphafetoprotein, human chorionic gonadotropin-beta, carcinoembryonic antigen, testosterone, prolactin, estradiol, luteinizing hormone, and follicle stimulating hormone were within normal limits. Ultrasonic examination showed a high echoic lesion in the right testis. Computerized tomography and magnetic resonance imaging showed no evidence of retroperitoneal lymph node enlargement. A high right orchiectomy was performed under a diagnosis of right testicular tumor. The right testis was elastic hard and measured 9 x 10 x 7 cm, weighing 450 g. The cut surface was light yellowish white and was completely displaced by the tumor. No normal tissue was seen. Histological examination showed a Sertoli cell tumor. No adjuvant therapy was performed. Neither recurrence nor evidence of metastasis has been detected for 8 years postoperatively. PMID- 10466071 TI - Cellular responses to FMLP challenging: a mini-review. AB - FMLP (N-formyl-methionyl-leucyl-phenylalanine) and other N-formylpeptides are powerful "activators" of polymorphonuclear and mononuclear phagocytes, but they are also active on other cell types. Present knowledge about formylpeptide receptors and the relevant tools for their imaging and the study of their dynamics are briefly discussed. The main responses elicited by FMLP in granulocytes are cell polarisation, the generation of reactive oxygen species, the production of arachidonic acid metabolites, and the release of lysosomal enzymes. The transduction cascades involved and the agents able to modulate these responses are reviewed. Homologous desensitization and heterologous desensitization of the FMLP-receptor following ligation of other chemokine receptors are also outlined. Finally, the receptor expression and the pharmacological and toxic actions of FMLP upon other tissues and organs, and its actions on the developing embryo, are illustrated. PMID- 10466072 TI - The effects of HIV viral load on the phagocytic activity of monocytes activated with lipopolysaccharide from oral microorganisms. AB - A study was undertaken to determine whether viral load status in HIV+ patients has any potential effect on monocyte phagocytic function both before and after challenge of the monocytes with lipopolysaccharide (LPS) isolated from oral microorganisms. LPS of two putative periodontal pathogens Porphyromonas gingivalis (P. gingivalis) and Fusobacterium nucleatum (F. nucleatum) was prepared. Whole blood samples in EDTA were collected from 30 HIV+ patients presenting for dental care at the University of Maryland. Control samples were prepared from appropriate uninfected individuals. Viral load was determined using quantitative RT-PCR (Amplicor, Roche Diagnostics). Phagocytic function was determined using FITC labeled Saccharomyces species in resting isolated monocytes and in cells after 24 h stimulation with 1 microgram/ml of LPS of P. gingivalis or F. nucleatum. Immunohistochemical staining was performed for complement receptor CR-1 (CD-35) on phagocyte cells. In HIV+ patients with high viral load (> 10,000 copies/ml), 13.5% of isolated resting monocytes demonstrated phagocytic activity, while 23% of the resting control monocytes from non-infected individuals showed phagocytic function. When the monocytes were stimulated with 1 microgram/ml of LPS of F. nucleatum, phagocytic activity was observed in 18.5% of monocytes in patients with high viral load, 33.5% with moderate viral load (400 10,00 copies/ml) and 51% with low viral load (<400 copies/ml), while 62% of the control monocytes demonstrated phagocytic activity. Stimulation of monocytes with LPS of P. gingivalis showed similar results. Complement receptor CD-35 showed a 50% decrease in expression in HIV+ patients with high viral load. A progressive decrease in monocyte/macrophage phagocytic function and CD-35 expression with and without oral LPS activation occurs after HIV infection and this trend appears to be accentuated in patients with high viral load. This relationship may contribute to increased susceptibility to oral opportunistic infections in advanced HIV+ patients. PMID- 10466073 TI - Protective effects of glucocorticoids on taxol-induced cytotoxicity in human leukemia HL-60 cells. AB - Although the therapeutic actions of glucocorticoids are largely attributed to anti-inflammatory and immunosuppressive effects, they have been implicated in enhancing tissue and cellular protections. In this study, we examined whether glucocorticoids including dexamethasone (Dex) and hydroxycortisone could diminish the cytotoxic effects of anti-microtubule agents including taxol (paclitaxel), microtubule stabilizing agent, and colchicine, microtubule disrupting agent, on human leukemia HL-60 cells. Taxol or colchicine decreased the viability of HL-60 cells in a dose-dependent manner. However, micromolar concentrations of glucocorticoids rendered HL-60 cells resistant against the cytotoxic activity of anti-microtubule agents. Pretreatments of the glucocorticoids were more effective than simultaneous treatments with antimicrotuble agents. The fact that actinomycin D or cycloheximide reversed the cytoprotective effects of glucocorticoids on cytotoxicities in HL-60 cells induced by antimicrotuble agents suggests glucocorticoids cytoprotection might be mediated via newly synthesized protein. Collectively, these data showed that micromolar concentrations of dexamethasone or hydrocortisone could attenuate the cytotoxic effects of taxol or colchicine on human leukemia HL-60 cells via protein synthesis. PMID- 10466074 TI - Retrospective study of adverse reactions to non steroid anti-inflammatory drugs (NSAIDs): predictive value of controlled challenge with alternative drugs. AB - Acetylsalicylic acid (ASA) and NSAIDs, which inhibit the cyclo-oxygenase enzyme (C-O), are responsible, when administered at therapeutic doses, for adverse reactions mainly involving the skin and respiratory tract. The prevalence of intolerance to ASA and NSAIDs, assessed by the Section of Allergic and Immunological Diseases at the University of Bari on a population of 15,800 patients referred for allergic diseases over a period of 7 years, was found to be 11.4%. The adverse reactions to NSAIDs observed were in most cases skin complaints (88.9%), followed by respiratory symptoms (asthma +/- rhinitis, rhinitis) and general symptoms (shock, hypotension, lipothymia). The most common types of NSAIDs taken were pyrazolones, salicylics, arylpropionics, paracetamol. Controlled oral challenges with alternative NSAIDs (especially nimesulide) confirm the predictive power of this test: in fact, among patients who showed tolerance to the challenge drug, only 10.6% manifested unexpected reactions during the course of one year's follow-up. PMID- 10466075 TI - Asiasari radix inhibits immunoglobulin E production on experimental models in vitro and in vivo. AB - Immunoglobulin (Ig) E is the principal Ig involved in immediate hypersensitivities and chronic allergic diseases. The hallmark of these disorders is increased IgE production. The effect of an aqueous extract of the roots of Asiasari radix (ARAE) on an in vivo and in vitro IgE production was investigated. ARAE dose-dependently inhibited the active systemic anaphylaxis and serum IgE production induced by immunization with ovalbumin, Bordetella pertussis toxin and aluminum hydroxide gel. ARAE strongly inhibited IL-4-dependent IgE production by lipopolysaccharide- stimulated murine whole spleen cells. In the case of U266 human IgE-bearing B cells, ARAE also showed an inhibitory effect on the IgE production. These results suggest that ARAE has an anti-allergic activity by inhibition of IgE production from B cells. PMID- 10466076 TI - Sequence determinants of modified cobra venom neurotoxin which induce immune resistance to experimental allergic encephalomyelitis: molecular mechanisms for immunologic action. AB - A nontoxic, iodoacetamide-modified cobratoxin derivative (CAM-NTX) induced resistance to experimental allergic encephalomyelitis (EAE) in guinea pigs. Resistance was retained after trypsin digestion and shown to reside in N-terminal and central peptides of CAM-NTX. A similarly modified protein cardiotoxin (CAM CTX), representative of proteins homologous with cobratoxin, was not immunosuppressive. Depressed clinical symptoms in EAE-resistant animals correlated with reduced lymphocytic infiltration of the brain. Antibody to myelin basic protein (MBP) was reduced in immunosuppressed animals. The immunoinhibitory determinants in CAM-NTX may mimic immune response suppressor proteins (SIRS-alpha 7) and the EAE-resistance region of MBP. PMID- 10466078 TI - Regulatory effects of cytokines and cyclosporine A on peripheral blood mononuclear cells from stable multiple sclerosis patients. AB - The proliferative response (PR) of peripheral blood mononuclear cells (PBMC) to lectins such as phytohemaglutinin (PHA), anti-CD3 monoclonal antibodies such as OKT-3 or phorbol esters such as tetradecanoyl-phorbol 13-acetate (TPA) was investigated in 18 stable multiple sclerosis (MS) patients (9 untreated and 9 treated patients) and 10 healthy controls. PBMC from untreated MS patients showed a significantly higher PR to PHA than healthy controls. The PR of PHA, anti-CD3 or TPA stimulated PBMC from treated patients was lower than that from untreated MS patients. Mitogen stimulated PBMC from untreated patients shown both increased sensitivity to the stimulatory effect of IL-2 and increased resistance to the inhibitory effect of IL-10 and IFN-alpha. The addition of IL-2 increased the PR in PHA-stimulated PBMC from untreated MS patients, but not in those from treated MS patients and healthy controls. Mitogen stimulated cells from untreated patients were more resistant to the inhibitory effect of IL-10 and IFN-alpha than PBMC from either treated MS patients or healthy controls. Cyclosporine A (CsA) inhibited the PR and the expression of activation antigens induced by PHA in PBMC from the three groups of subjects. This inhibitory effect of CsA have was enhanced by the addition of IFN-alpha. PMID- 10466077 TI - Modulating effects of sensory and autonomic neuropeptides on murine splenocyte proliferation and cytokine secretion induced by Leishmania major. AB - The intimate, bidirectional link between neuroendocrine and immune systems is now accepted. A modulating effect of the nervous system on immune and inflammatory responses has been corroborated by identification of neuropeptide receptors on immunocompetent cells and the finding that neuropeptides can regulate leukocyte functions. The present study was undertaken to investigate the possible immunomodulatory role of sensory (SOM, CGRP and SP) and autonomic (VIP and NPY) neuropeptides in a murine model of cutaneous leishmaniasis, using two genetically different inbred mouse strains, BALB/c and C57BL/6, respectively susceptible and resistant to Leishmania (L.) major infection. The parameters studied were extent of splenocyte proliferation, as measured by thymidine uptake, and the ability of these cells to secrete IFN-gamma and IL-4 by using a two-site ELISA, upon in vitro challenge with L. major parasites and addition of the neuropeptides. The resistant mouse splenocyte proliferation was enhanced by SOM, CGRP, and VIP at 10(-5), 10(-6) and 10(-9) M concentration, respectively, but was inhibited by NPY at 10(-5) M. Proliferation of the splenocytes from the susceptible strain was inhibited by SOM (10(-11) M) and CGRP(10(-5) M). Somatostatin, at various concentrations, stimulated IFN-gamma secretion in both mouse strain splenocytes, and IL-4 production in the susceptible mouse. Calcitonin gene-related peptide enhanced IFN-gamma secretion in susceptible mouse splenocytes at 10(-6), 10(-7) and 10(-9) M, as did VIP at 10(-10) M and NPY at 10(-7) M. Vasoactive intestinal peptide also stimulated IL-4 production in BALB/c splenocytes at all concentrations used. Substance P had no effect on either cell proliferation or cytokine secretion in either of the two mouse strains. These findings indicate that the nervous system, represented by sensory and autonomic nerve terminals and their content of neuromediators, may be involved in the pathophysiology of cutaneous leishmaniasis. PMID- 10466079 TI - Evaluation of nonspecific immunity and plasma levels of interferon-gamma, interleukin-6 and tumor necrosis factor-alpha in preeclampsia. AB - In this study we evaluated the maternal cell-mediated immune aspects of preeclampsia in terms of phagocytosis and killing of monocytes and polymorphonuclear cells. To evaluate the contribution of cytokines (Cks) in the pathophysiology of preeclampsia, we investigated the plasma levels of interferon gamma (IFN-gamma), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF alpha), respectively, by an enzyme-linked immunosorbent assay. Data showed that phagocytic and killing activities of monocytes were depressed in preeclamptic and normal pregnancies. At the same time, IFN-gamma plasma levels were undectable in both groups. Conversely, we detected significant levels of TNF-alpha in plasma from preeclamptic and normal pregnancies. Moreover, since in three preeclamptic patients the onset of severe preeclampsia was associated with a sharp increased of TNF-alpha plasma levels, we suggest that an increased production of this CK may be implicated in the pathophysiology of preeclampsia. PMID- 10466080 TI - CD69 and regulation of the immune function. AB - CD69, also known as activation inducer molecule, very early activation antigen, MLR-3 and Leu-23, is a member of the natural killer (NK) cell gene complex family of signal transducing receptors. CD69 is as a type II transmembrane glycoprotein with a C-type lectin binding domain in the extracellular portion of the molecule. CD69 expression is induced in vitro on cells of most hematopoietic lineages, including T and B lymphocytes, NK cells, murine macrophages, neutrophils and eosinophils, while it is constitutively expressed on human monocytes, platelets and epidermal Langerhans cells. Although a specific ligand for CD69 has not been identified, its wide cellular distribution and the induction of intracellular signals upon CD69 crosslinking suggest a role for the receptor in the biology of hematopoietic cells. Moreover, certain results indicate that CD69 may be involved in the pathogenesis of such diseases as rheumatoid arthritis, chronic inflammatory liver diseases, mild asthma, and acquired immunodeficiency syndrome. PMID- 10466081 TI - Effects of glyburide-cyclosporin A interaction on interleukin-2 production in rats. AB - The effects of simultaneous administrations of Cyclosporin A (CsA) and Glyburide on the immune system of rats has been evaluated in terms of Interleukin-2 (IL-2) production by Concanavalin A (ConA) stimulated splenocytes and exogenous IL-2 binding capacity. The inhibitory effect of Cyclosporin A on IL-2 production of lymphoid cells is well known. Spleen cells from rats receiving CsA had reduced levels of IL-2 when compared to untreated controls or rats receiving Glyburide only. Splenocytes from rats receiving both drugs had reduced levels of IL-2 when they were sacrificed 24 hours after one or three CsA administrations; instead when the animals were sacrificed 6 days after three CsA administrations, their ability of producing IL-2 is increased as well as increasing exogenous IL-2 binding capacity. These findings let us hypothesize that when there are lower concentrations of CsA in lymphocytes there is an increase of cellular metabolism induced by Glyburide that leads to an increase in IL-2 secretion and in IL-2 receptor expression on cellular surface restoring these levels to normal or slightly above normal levels. PMID- 10466082 TI - In vitro effects of various metals on natural killer cell activity in cultured human lymphocytes. AB - Heavy metals have been shown to have a differential effects on various aspects of immune response. Recently natural killer cells have been widely investigated due to their purported role in immune surveillance. To ascertain the immunotoxic effects of lead, cadmium, nickel and chromium on natural killer (NK) cell activity in vitro, peripheral blood lymphocytes from normal donors were examined in the presence of different concentrations (10(-5)-10(-8) M) of four selected metal salts (cadmium sulphate, lead nitrate, chromium nitrate and nickel sulphate). NK cell activity was evaluated in a 4-h chromium release assay against K562 target cells. All of the metal salts were found to exert no effect on NK cell function in the human concentration range. PMID- 10466083 TI - The effects of Chlorella vulgaris in the protection of mice infected with Listeria monocytogenes. Role of natural killer cells. AB - In this work we have demonstrated the effects of oral administration of Chlorella vulgaris (CV) on Natural Killer cells (NK) activity of mice infected with a sublethal dose of viable Listeria monocytogenes. The treatment with C. vulgaris produced a significant increase on NK cells activity in normal (non-infected) animals compared to the animals that received only vehicle (water) (p < 0.0001). Similarly, the infection alone produced a significant increase on NK cells activity, which was observed at 48 and 72 hours after the inoculation of L. monocytogenes. Moreover, when CV was administered in infected animals, there was an additional increase in NK cells activity which was significantly higher than that found in the infected groups (p < 0.0001). CV treatment (50 and 500mg/Kg) of mice infected with a dose of 3 x 10(5) bacteria/animal, which was lethal for all the non-treated controls, produced a dose-response protection which led to a 20% and 55% survival, respectively (p < 0.0001). PMID- 10466084 TI - Measurement of the respiratory burst and chemotaxis in polymorphonuclear leukocytes from anti-ChE insecticides-exposed workers. AB - Neutrophil function in 32 workers occupationally exposed to anti-ChE insecticides, as measured by chemotaxis through the leading front method and nitroblue tetrazolium dye reduction, was investigated and compared to those of age- and sex-matched controls. The cholinesterase (ChE) activity was normal in all the workers studied, although decrease of chemotaxis and of nitroblue tetrazolium reduction was observed in the exposed population. These results suggest that the identified functional changes in polymorphonuclear neutrophils might be an early indicator of anti-ChE insecticides toxicity, even in those individuals with no impairment in the ChE activity. PMID- 10466085 TI - Action of Sosiho-Tang on systemic and local anaphylaxis by anal administration. AB - The herbal formulation Soshiho-Tang (SS-Tang) has been used against allergic disease for generations, and still occupies an important place in traditional medicine in Korea. Previously, we reported that SS-Tang potently inhibited mast cell- mediated anaphylaxis when orally administered. In this study, we investigated the effect of SS-Tang by anal administration in anaphylaxis responses. SS-Tang dose-dependently inhibited compound 48/80-induced systemic anaphylaxis with doses of 10(-4) to 1 g/kg 1 h before anally administered. Of special note, SS-Tang inhibited systemic anaphylaxis completely with a dose of 1 g/kg. SS-Tang reduced plasma histamine levels induced by compound 48/80 significantly. However, the mortality was 100% when SS-Tang was administered after compound 48/80 treatment. SS-Tang (10(-1) g/kg) also inhibited passive cutaneous anaphylaxis activated by anti-dinitrophenyl IgE antibody by 30.9%. These results provide evidence that anal therapy of SS-Tang may be beneficial in the treatment of systemic and local anaphylaxis. PMID- 10466086 TI - [Percutaneous transluminal coronary angioplasty for restenosis following Palmaz Schatz stenting: larger size balloons are more effective?]. AB - Intracoronary stenting reduces the restenosis rate after percutaneous transluminal coronary angioplasty (PTCA). However, restenosis still occurs in 20% to 30% of patients after stenting. Management of in-stent restenosis has become a significant challenge in interventional cardiology. The efficacy of re-PTCA with a larger balloon was investigated for restenosis following Palmaz-Schatz stenting. Clinical and angiographic results were compared in 46 consecutive patients with in-stent restenosis after one Palmaz-Schatz stenting. Twenty patients underwent redilation with a slightly larger balloon than used at the stenting (Large group) and 26 underwent redilation with the same size balloon as at the stenting (Control group). The clinical factors, lesion characteristics, lesion length, reference diameter and minimal luminal diameter at re-PTCA for the in-stent restenosis did not differ significantly between the 2 groups. Minimal luminal diameter of the Large group after re-PTCA was significantly larger than that of the Control group (3.1 +/- 0.2 vs 2.9 +/- 1.2 mm, p < 0.05). Follow-up angiography showed that the minimal luminal diameter of the Large group was significantly larger (2.1 +/- 0.6 vs 1.7 +/- 0.6 mm, p < 0.05) and the re restenosis rate of Large group tended to be lower than that of the Control group (15% vs 38%, p = 0.07). Re-PTCA for in-stent restenosis with a slightly larger balloon than used at the stenting reduces the re-restenosis rate. PMID- 10466087 TI - [Relationship between ventricular arrhythmias and myocardial fatty acid metabolism in patients with coronary heart disease: evaluation using iodine-123 beta-methyl-p-iodophenyl-pentadecanoic acid]. AB - The effect of metabolic abnormalities of myocardial fatty acids on ventricular arrhythmias was evaluated by myocardial imaging with iodine-123 beta-methyl-p iodophenyl-pentadecanoic acid (BMIPP) in 27 patients with coronary heart disease. The disturbance of myocardial blood flow was also evaluated using thallium-201 (Tl). The patients were divided into 2 groups based on the character of the premature ventricular contractions: Group A: number of contractions > or = 120 per day and/or consecutive contractions (n = 9, mean age 63.7 yr), and Group B, number of contractions < 120 per day and no consecutive contractions (n = 18, mean age 64.2 yr). Left ventricular ejection fraction was measured by left ventriculography, and significant coronary artery stenosis was defined as stenosis of 75% or greater. Cardiac scintigraphy was performed using single photon emission computed tomography with BMIPP at rest in 27 patients and in the early phase (early Tl) and delayed phase of Tl (delayed Tl) in 20 patients. BMIPP and Tl uptakes were scored as 0: absent, 1: moderately reduced, 2: mildly reduced and 3: normal in 7 segments of the left ventricular wall and then the total scores were calculated in each patient. Ejection fraction significantly correlated with the scores of BMIPP, and early and delayed Tl(p < 0.001, respectively), although the ejection fraction in Group A was significantly less than in Group B (51.2 +/- 16.7% vs 68.2 +/- 14.4%, p < 0.02). The BMIPP scores in Group A were significantly less than those in Group B (14.2 +/- 4.3 vs 17.2 +/- 3.1, p < 0.05), but the early and delayed Tl scores in Group A were not significantly different compared with those in Group B. The BMIPP scores showed no significant differences between the patients with and without significant coronary artery stenosis, but the early and delayed Tl scores in the patients with stenosis were significantly less than those in patients without stenosis (early Tl: 19.8 +/- 2.6 vs 16.8 +/- 2.8, p < 0.01; delayed Tl: 19.6 +/- 2.5 vs 16.8 +/- 3.1, p < 0.05). More patients had values of Tl minus BMIPP scores of at least 3 in Group A than in Group B (early Tl-BMIPP > or = 3: 66.7% vs 7.7%, p < 0.01; delayed Tl-BMIPP > or = 3: 50.0% vs 7.7%, p < 0.05). In patients with coronary heart disease, decreased ejection fraction is thought to be a major cause of ventricular arrhythmias, and also associated with abnormal myocardial metabolism and abnormal myocardial blood flow. Furthermore, ventricular arrhythmias were more closely correlated with abnormal metabolism of fatty acids than with disturbance of blood flow in the myocardium, and large mismatching between the 2 images may be related to risky ventricular arrhythmias in coronary heart disease. PMID- 10466088 TI - [Clinical outcome of catheter fragmentation and aspiration therapy in patients with acute pulmonary embolism]. AB - The clinical usefulness of catheter fragmentation and aspiration therapy was studied in 8 patients with acute pulmonary embolism who received thrombolytic therapy using urokinase or tissue-type plasminogen activator (t-PA) (thrombolysis group) and 8 patients who underwent catheter fragmentation and aspiration therapy using a percutaneous transluminal coronary angioplasty (PTCA) guide catheter (catheter group). The patients were selected from 20 patients with a definite diagnosis of acute pulmonary embolism based on pulmonary arteriography and nuclear imaging. Urokinase (48 x 10(4) to 96 x 10(4) unit/day) or t-PA (12 x 10(6) unit/day) was administered intravenously for mean 4 days in the thrombolysis group. Pulmonary artery pressure was first measured using a Swan Ganz catheter via the jugular vein or the femoral vein in the catheter group. Then, a PTCA guide catheter was advanced into the pulmonary artery, and the thrombus was disrupted repeatedly using a Radifocus wire, followed by manual aspiration. Subsequent treatment consisted of intravenous infusion of heparin (10,000 to 15,000 unit/day) and urokinase (24 x 10(4) to 48 x 10(4) unit/day) for mean 6 days. Partial revascularization was achieved in all patients in both groups. Five patients in the thrombolysis group died within 1 month due to respiratory failure, re-embolization, and/or hemorrhagic complications. One patient in the catheter group died of hemorrhagic shock. Pulmonary artery systolic pressure in the catheter group was significantly reduced from 47.4 to 26.5 mmHg (p < 0.01). Catheter treatment of acute pulmonary embolism associated with acute circulatory failure such as shock can lead to rapid hemodynamic improvement. In contrast, thrombolysis is an effective treatment, but bleeding problems are common and caution is required. Catheter fragmentation and aspiration therapy is effective for acute pulmonary embolism, is minimally invasive, and should be considered the treatment of first choice. PMID- 10466089 TI - [Effects of low dose pimobendan in patients with cor-pulmonale]. AB - The acute and chronic efficacy of low dose pimobendan (1.25 mg x 2/day) was evluated in patients with cor-pulmonale. Fifteen patients (12 men, 3 women, mean age 73 +/- 5 yr) with right ventricular dysfunction judged by Tei's Doppler index (> or = 0.4) and poor working capacity (exercise tolerance: 2.2-6.6 MET) were studied. Mean pulmonary artery pressure, cardiac output, total pulmonary resistance using Swan-Ganz catheter, and arterial oxygen and carbon dioxide pressure (PaO2, PaCO2) were measured before and 24 hr after pimobendan administration. Maximal oxygen intake (MET), saturation of arterial blood oxygen at rest and desaturation by treadmill stress test were measured before and 1 month after pimobendan administration. Pulmonary artery pressure decreased (17.6 +/- 4.7 to 10.2 +/- 2.3 mmHg, p < 0.001) and cardiac output increased (3.5 +/- 0.6 to 5.1 +/- 0.9 l/min, p < 0.001), resulting in decreased total pulmonary resistance (5.0 +/- 1.3 to 2.1 +/- 0.7 U, p < 0.001), and a mild decrease in PaO2 (74 +/- 8 to 70 +/- 10 mmHg, p < 0.05). Exercise tolerance improved significantly (4.8 +/- 1.7 to 6.8 +/- 2.2 MET, p < 0.001), without deterioration of PaO2 and desaturation. These results indicate that low dose pimobendan is useful for the treatment of patients with cor-pulmonale. PMID- 10466090 TI - [Cardiac sarcoidosis complicated by multivessel coronary spasm: a case report]. AB - A 63-year-old woman with abnormal Q waves in leads II, III, aVF developed ventricular tachycardia after an operation for thyroid carcinoma. Coronary arteriography revealed no organic stenosis, but acetylcholine induced total occlusion of the right coronary artery and severe narrowing of the left coronary artery. Left ventriculography showed inferoposterior and septal akinesis, and echocardiography revealed slight thinning of these affected walls. She had old myocardial infarction due to spasm. One year later, she developed bilateral uveitis and recurrence of tachycardia. Cardiac sarcoidosis was diagnosed by endomyocardial biopsy. In our patient with cardiac sarcoidosis, the presence of multivessel coronary spasm made the diagnosis difficult and may have contributed to cardiac dysfunction. Coronary arteries are rarely involved, but the development of coronary spasm may be linked to sarcoidosis. PMID- 10466091 TI - [A 50-year-old woman complaining of severe dyspnea]. PMID- 10466092 TI - [Throat discomfort developing immediately after left ventriculography]. PMID- 10466093 TI - Language and the infant brain. AB - Three logically and empirically independent issues are often conflated in theory and research on brain and language: localization, innateness, and domain specificity. Research on adults and infants with focal brain injury support the following conclusions: (a) linguistic knowledge is not innate, and it is not localized in a clear and compact from in either the infant or adult brain; (b) the infant brain is not, however, a tabala rasa-it is already highly differentiated at birth, and certain regions are biased from the beginning toward modes of information processing that are particularly useful for language, leading (in the absence of local injury) to the standard form of brain organization for language; (c) the processing biases that lead to the "standard brain plan" are innate and localized, in both infants and adults, but they are not specific to language; and (d) the infant brain is highly plastic, permitting alternative "brain plans" for language to emerge if the standard situation does not hold. PMID- 10466094 TI - Narrowing the distance to language: one step at a time. AB - Infants' earliest attempts at word segmentation appear to be guided by a single source of information (e.g., English-learners initially rely on the predominant stress pattern of words). This initial strategy successfully identifies many potential words in the input, but mis-segments others. However, simply breaking the input into smaller chunks helps learners to identify other possible cues to the location of word boundaries in utterances. Because no one source of information is completely reliable, listeners must eventually rely on multiple cues to segment words. The development of such skills is not critical for developing a native language vocabulary, but also for acquiring the grammatical organization of utterances. Tracking familiar sound patterns, such as function words and grammatical morphemes, may help in learning about syntactic organization. One factor that facilitates learning about the distribution of such elements is sensitivity to boundaries of prosodic phrases. Access to such linguistically-relevant chunks also helps in tracking the distribution of words in the input. PMID- 10466095 TI - Precursors to speech in infancy: the prediction of speech and language disorders. AB - During the canonical stage of infant babbling, infants produce well-formed syllables, often in reduplicated sequences such as "bababa." Although nearly all infants with normal hearing begin the canonical stage by 10 months of age, a few are delayed, and these infants may be of special interest. Recent studies indicate that late onset of canonical babbling may be a predictor of disorders. A simple screening procedure that focuses on canonical babbling was used to evaluate over 3400 infants at risk who were about 10 months of age. Among infants who showed late onset of canonical babbling, fewer than half had been previously diagnosed as having a significant medical problem that might have accounted for the delay. A follow-up study indicated that infants with delayed canonical babbling had smaller production vocabularies at 18, 24, and 30 months than did infants in the control group. The results suggest that late onset of canonical babbling, a factor that can be monitored effectively through an interview with a parent, can predict delay in the onset of speech production. PMID- 10466096 TI - Discussion: early speech perception and production. PMID- 10466098 TI - From the big bang to the brain. AB - Current research on the capacities of the infant has lead to a better understanding of developmental processes underlying cognition and motor skill acquisition. ASHA's Eighth Annual Research Symposium on Infant-Toddler Development, in November 1998, included a presentation on developmental cognitive science by Dr. Andrew Meltzoff and a presentation on motor skill acquisition by Dr. Esther Thelen. The theoretical constructs and data presented served to broaden our current perspectives on infant abilities. The data reported by Meltzoff and Thelen challenged several long-standing theories of infant cognition and motor development. Alternative theoretical models were used to describe skill acquisition during the first several years of life. Our response will include a brief summary of each investigator's presentation, discuss their findings with respect to research in the area of infant speech physiology and production, and provide possible future directions and challenges for individuals conducting developmental research. PMID- 10466097 TI - Origins of theory of mind, cognition and communication. AB - There has been a revolution in our understanding of infant and toddler cognition that promises to have far-reaching implications for our understanding of communicative and linguistic development. Four empirical findings that helped to prompt this change in theory are analyzed: (a) Intermodal coordination--newborns operate with multimodal information, recognizing equivalences in information across sensory-modalities; (b) Imitation--newborns imitate the lip and tongue movements they see others perform; (c) Memory--young infants form long-lasting representations of perceived events and use these memories to generate motor productions after lengthy delays in novel contexts; (d) Theory of mind--by 18 months of age toddlers have adopted a theory of mind, reading below surface behavior to the goals and intentions in people's actions. This paper examines three views currently being offered in the literature to replace the classical framework of early cognitive development: modularity-nativism, connectionism, and theory-theory. Arguments are marshaled to support the "theory-theory" view. This view emphasizes a combination of innate structure and qualitative reorganization in children's thought based on input from the people and things in their culture. It is suggested that preverbal cognition forms a substrate for language acquisition and that analyzing cognition may enhance our understanding of certain disorders of communication. PMID- 10466103 TI - Effects of synthetic pyrethroids and methidation on activities of some digestive enzymes in carp (Cyprinus carpio L.). AB - The effects of pyrethroid pesticides (deltamethrin, permethrin and cypermethrin) and an organophosphate ester (methidation) on the activities of carp trypsin, alpha-chymotrypsin, carboxypeptidase A and lipase were studied. The enzymes were isolated from the gastrointestinal tract and the effects of the pesticides were investigated during incubation for 5 min. The activity of trypsin was influenced only slightly by the presence of deltamethrin and methidation, whereas permethrin and cypermethrin caused significant inhibition. The pyrethroid pesticides at lower concentrations resulted in a slight activation of alpha-chymotrypsin. Methidation inhibited the alpha-chymotrypsin activity by about 20%. These pesticides modified the lipase activity to a lesser extent; the highest inhibition was measured with cypermethrin. The carboxypeptidase A activity was inhibited by both pyrethroid pesticides and methidation. The results suggest that these pesticides might interact with the active conformation of the studied hydrolytic enzymes, resulting in changes in their activities. PMID- 10466104 TI - Determination of organochlorine and organophosphate pesticide residues in fruits, vegetables and sediments. AB - A method is described for the determination of organochlorine and organophosphate pesticide residues in fruits, vegetables and sediments. The concentrated solvent extract was sealed in a polymeric membrane tube, dialysed in cyclohexane and the solvent replaced with hexane. The organophosphates were analysed on a specific thermionic detector without further clean-up. For the organochlorine pesticides the extract was eluted through 3 g of alumina and analysed on GC/ECD. The clean up for sediment extract was carried out on a 10 g alumina column with 100 mL hexane containing 5% acetone and the eluate was concentrated to 5 mL. The detection limit for organophosphates on a 40 g sample and a final volume of 10 mL was on the average 0.01 mg/kg. The detection limit for organochlorine pesticides, with the final volume of 25 mL, was 0.005 mg/kg for all pesticides except for p,p'-DDT and endosulfan sulphate, which was 0.01 mg/kg. The detection limit for organochlorine pesticides in sediment, with the final volume of 2 mL, was less than 1 microgram/kg and for organophosphate pesticides less than 10 micrograms/kg when the final volume was made to 0.5 mL. At the detection limits the method produced a very high coefficient of variation for both organochlorine and organophosphate pesticides. PMID- 10466105 TI - Comparison of gas chromatography and immunoassay methods in quantifying fenitrothion residues in grape juice processed into alcoholic drinks. AB - Wine and Arak, the national alcoholic drink in Lebanon, were prepared from grape juice fortified with fenitrothion to a concentration of 20ppm. Samples of the 11 fractions produced by the fermentation and distillation steps were analyzed for fenitrothion residues using gas chromatography (GC) and enzyme-linked immunosorbent assay (ELISA). Results of residue analyses showed that the two techniques were highly correlated (r = 0.978) and indicated that fenitrothion was stable during the fermentation steps but not during distillation. The clarified wine 35 days later contained about 85% (15.3 ppm) of the fenitrothion concentration found in the juice as determined by GC analysis. Arak was prepared by a two-steps distillation of the clarified wine. The alcohol distillate and undistilled fraction from the first distillation contained 2.5 ppm and 5.8 ppm of fenitrothion, respectively. No fenitrothion residues were detected by both techniques in the four fractions collected from the second distillation step. PMID- 10466107 TI - Sub-chronic effect of neem based pesticide (Vepacide) on acetylcholinesterase and ATPases in rat. AB - Acetylcholinesterases (AChE), Na(+)-K+, Mg2+ and Ca(2+)-ATPases were monitored in rat brain when treated orally with 80, 160 and 320 mg/kg of Vepacide, an active ingredient from neem seed oil, daily for 90 days. Brain AChE, Na(+)-K+ and Ca(2+) ATPases were inhibited whereas Mg(2+)-ATPase levels were enhanced in both the sexes after 45 and 90 days of treatment. The relative sensitivities of these ATPases to Vepacide indicated that Ca(2+)-ATPase being more sensitive than Na(+) K(+)-ATPase in both the sexes. The magnitude of Ca(2+)-ATPase inhibited by this compound was higher than that of brain AChE. It appears to be sexual dimorphism in the alterations of brain AChE, Na(+)-K+ and Mg(2+)-ATPases by Vepacide with females being significant when compared with males. After 28 days of post treatment the alterations observed were approached to those of controls both in male and female rats showing reversal of the toxicity. These results indicated that the ATPases were potently inhibited by Vepacide and seemed to be its precise target among the enzyme studied. This can be used as biochemical marker of exposure to this neem derived product. PMID- 10466108 TI - Radionuclides and trace elements in fish collected upstream and downstream of Los Alamos National Laboratory and the doses to humans from the consumption of muscle and bone. AB - The purpose of this study was to determine radionuclide and trace element concentrations in bottom-feeding fish (catfish, carp, and suckers) collected from the confluences of some of the major canyons that cross Los Alamos National Laboratory (LANL) lands with the Rio Grande (RG) and the potential radiological doses from the ingestion of these fish. Samples of muscle and bone (and viscera in some cases) were analyzed for 3H, 90Sr, 137Cs, totU, 238Pu, 239,240Pu, and 241Am and Ag, As, Ba, Be, Cr, Cd, Cu, Hg, Ni, Pb, Sb, Se, and Tl. Most radionuclides, with the exception of 90Sr, in the muscle plus bone portions of fish collected from LANL canyons/RG were not significantly (p < 0.05) higher from fish collected upstream (San Ildefonso/background) of LANL. Strontium-90 in fish muscle plus bone tissue significantly (p < 0.05) increases in concentration starting from Los Alamos Canyon, the most upstream confluence (fish contained 3.4E-02 pCi g-1 [126E-02 Bq kg-1]), to Frijoles Canyon, the most downstream confluence (fish contained 14E-02 pCi g-1 [518E-02 Bq kg-1]). The differences in 90Sr concentrations in fish collected downstream and upstream (background) of LANL, however, were very small. Based on the average concentrations (+/- 2SD) of radionuclides in fish tissue from the four LANL confluences, the committed effective dose equivalent from the ingestion of 46 lb (21 kg) (maximum ingestion rate per person per year) of fish muscle plus bone, after the subtraction of background, was 0.1 +/- 0.1 mrem y-1 (1.0 +/- 1.0 microSv y-1), and was far below the International Commission on Radiological Protection (all pathway) permissible dose limit of 100 mrem y-1 (1000 microSv y-1). Of the trace elements that were found above the limits of detection (Ba, Cu, and Hg) in fish muscle collected from the confluences of canyons that cross LANL and the RG, none were in significantly higher (p < 0.05) concentrations than in muscle of fish collected from background locations. PMID- 10466109 TI - Radionuclides in deer and elk from Los Alamos National Laboratory and the doses to humans from the ingestion of muscle and bone. AB - This paper summarizes radionuclide concentrations (3H, 90Sr, 137Cs, 238Pu, 239,240Pu, 241Am, and totU) in muscle and bone tissue of mule deer (Odocoileus hemionus) and Rocky Mountain elk (Cervus elaphus) collected from Los Alamos National Laboratory (LANL), Los Alamos, New Mexico, lands from 1991 through 1998. Also, the committed effective dose equivalent (CEDE) and the risk of excess cancer fatalities (RECF) to people who ingest muscle and bone from deer and elk collected from LANL lands were estimated. Most radionuclide concentrations in muscle and bone from individual deer (n = 11) and elk (n = 22) collected from LANL lands were either at less than detectable quantities (where the analytical result was smaller than two counting uncertainties) and/or within upper (95%) level background (BG) concentrations. As a group, most radionuclides in muscle and bone of deer and elk from LANL lands were not significantly higher (p < 0.10) than in similar tissues from deer (n = 3) and elk (n = 7) collected from BG locations. Also, elk that had been radio collared and tracked for two years and spent an average time of 50% of LANL lands were not significantly different in most radionuclides from road kill elk that have been collected as part of the environmental surveillance program. Overall, the upper (95%) level net CEDEs (the CEDE plus two sigma for each radioisotope minus background) at the most conservative ingestion rate (50 lbs of muscle and 13 lbs of bone) were as follows: deer muscle = 0.22 mrem y-1 (2.2 microSv y-1), deer bone = 3.8 mrem y-1 (38 microSv y-1), elk muscle = 0.12 mrem y-1 (1.2 microSv y-1), and elk bone = 1.7 mrem y-1 (17 microSv y-1). All CEDEs were far below the International Commission on Radiological Protection guideline of 100 mrem y-1 (1000 microSv y 1), and the highest muscle plus bone net CEDE corresponded to a RECF of 2E-06, which is far below the Environmental Protection Agency upper level guideline of 1E-04. PMID- 10466111 TI - Vascular endothelial growth factor and microvascular permeability. AB - Vascular Endothelial Growth Factors (VEGFs) are endogenously produced vascular cytokines which result in angiogenesis, vasodilatation, and increased microvascular permeability in vivo. They are endothelial specific and result in mitosis, migration, stress fiber formation and increased permeability of endothelial cells in culture. They have been critically implicated in a host of pathological conditions including solid tumor growth and diabetes, and been proposed as a therapy for coronary and peripheral ischemic disease. However, the potent permeability-enhancing properties of VEGFs are very poorly understood. The pharmacology, signal transduction pathways, intracellular signaling mechanisms, and ultrastructural changes which result in increased permeability are still not clear. This review discusses the available evidence for how VEGFs increase permeability in vivo, and some of the pitfalls in interpreting experiments which do not take into account the vasoactive properties of VEGFs. It also discusses the clinical implications of the permeability enhancing effect of VEGFs, and the relevance of these studies to development of new therapies. PMID- 10466112 TI - Microcirculatory stasis induced by hemorheological disorders: further evidence. AB - OBJECTIVE: Reinvestigate the microcirculatory alterations immediately responsible for blood rheological disorders and blood stases, which are related to red blood cell (RBC) aggregation in capillaries. METHODS: Blood rheological disorders were produced by significantly intensified intravascular red blood cell aggregation in the intestinal mesentery of Wistar rats and in the cerebral cortex of Chinchilla rabbits, either systemically (by intravascular administration of high molecular weight dextran) or locally (by increase of high-molecular compounds in blood plasma inside individual or groups of capillaries). RESULTS: Under conditions where the microvascular lumina were not decreased and the arteriolovenular pressure gradients got even higher, the significantly enhanced intravascular RBC aggregation resulted in the slowing down of blood flow in the microvessels to a full stop. CONCLUSION: A significant increase in microvascular RBC aggregation results in local hemorheological disorders, which is, in all probability, related to derangement of the blood-flow structuring in microvessels. PMID- 10466113 TI - Effect of reactive oxygen metabolites on endothelial permeability: role of nitric oxide and iron. AB - OBJECTIVE: We evaluated the effects of the xanthine oxidase (XO)-derived reactive oxygen metabolites on the permeability of bovine pulmonary artery-endothelial monolayers and examined how iron and nitric oxide (NO) participate in these changes in permeability. METHODS: Permeability was measured using a cell-column chromatographic method in which monolayers were exposed to combinations of agents. RESULTS: Exposure of monolayers to a superoxide/peroxide generator, xanthine (X, 0.1 mM)/XO (25 mU/mL), increased solute permeability after 10 minutes, but the same dose of either X or XO alone did not. Exposure of monolayers to peroxide (0.1 mM) also increased permeability, but only after 70 minutes. This X/XO permeability was attenuated by either catalase, superoxide dismutase, methionine (1 mM), an oxy-radical scavenger, or desferrioxamine (0.1 mM), an iron chelator. Spermine NONOate (SNO), an NO donor, attenuated X/XO permeability at 0.1 mM, but this protection was not significant at 0.01 or 1 mM. Spermine NONOate (0.1 mM) did not alter the permeability produced by 0.1 mM peroxide. L-N5-(1-iminoethyl)-ornithine (10 microM), an NO synthase inhibitor, completely blocked peroxide-, and partially attenuated X/XO-mediated permeability. However, 3-morphosynodiomine (SIN-1, 1 mM) plus catalase (1,000 U/mL), a peroxynitrite generator, did not alter permeability. CONCLUSIONS: Xanthine/Xanthine Oxidase permeability involves peroxide, superoxide, oxy radicals, and iron. Endogenous NO may regulate peroxide-, but not superoxide mediated permeability. The protective effects of exogenous NO on the X/XO permeability may represent interactions between superoxide, peroxide, and cell surface-bound iron. PMID- 10466114 TI - Effect of concentration and hyaluronidase on albumin diffusion across rabbit mesentery. AB - OBJECTIVE: To measure the diffusion coefficient of albumin through rabbit mesentery using the steady-state flux of radioactive tracer 125I-albumin. The effect of albumin concentration and testicular hyaluronidase were also studied. METHODS: Mesenteric tissue was bonded between two plates, exposing a 7 mm diameter surface, with two chambers on either side. One chamber was filled with a test solution of albumin containing the radioactive tracer and the other with lactated Ringer solution. The solutions in both chambers were stirred with small magnetic cylinders. The chamber filled with lactated Ringer solution was placed in a well-type NaI(Tl) detector, and the radiation emitted from the tracer that diffused across the mesentery was monitored continuously for 9 hours. The diffusion coefficient (D) was calculated using Fick's law of diffusion. The diffusion coefficient was measured at albumin concentration differences (delta C) between approximately 0 and 10 g/dL. The diffusion coefficient was also measured with testicular hyaluronidase at three different albumin-concentration differences. RESULTS: The diffusion coefficient increased significantly (P < 0.0001) approximately three-fold from a mean value of 2.2 x 10(-8) +/- 1.2 x 10( 8) (SD) cm2/s at 0-0.5 g/dL delta C to 5.9 x 10(-8) +/- 1.1 x 10(-8) (SD) cm2/s at 10 g/dL delta C. The values are much less than the free diffusion coefficient of albumin (6 x 10(-7) cm2/s). Testicular hyaluronidase added to the albumin solution decreased D by approximately 60%, but did not eliminate the increase in D with delta C. CONCLUSIONS: The increase in D with delta C and the reduced D with hyaluronidase were attributed to a reduced albumin-excluded volume caused by an interaction between albumin and hyaluronan. Further studies are required to define this interaction. PMID- 10466115 TI - Endometrial endothelial cells express estrogen and progesterone receptors and exhibit a tissue specific response to angiogenic growth factors. AB - OBJECTIVE: To develop a reliable method for the isolation and longterm culture of microvessel endothelial cells from human endometrium and to evaluate their response to angiogenic growth factors and steroid hormones in comparison to endothelial cells derived from other organs. METHODS: Endometrial tissue from hysterectomy specimens were digested sequentially with collagenase and trypsin, cultured for 24 h, then selected by adhesion to anti-CD-34 coated magnetic beads. Alternatively, anti-CD-34-coated beads could also be substituted by Ulex europaeus agglutinin-1, anti-PECAM, or anti-E-selectin-coated beads. Characterization of the isolated cultures included expression of endothelial cell markers, regulation of E-selectin in response to TNF-alpha, proliferative response to angiogenic growth factors, and expression of progesterone and estrogen receptors. We also analyzed the relative binding affinity of VEGF on endometrial endothelial cells in comparison to other endothelial cell types. RESULTS: Selection on anti-CD-34-coated beads eliminated contaminating cells and resulted in a homogeneous population of human endometrial endothelial cells (HEEC), as assessed by expression of PECAM, von Willebrand's factor, and uptake of acetylated-LDL. HEEC also upregulated E-selectin in response to TNF-alpha in a manner similar to that seen for other endothelial cell types. Expression of progesterone and estrogen receptor was revealed by immunocytochemistry and RT-PCR consistently until passage 5. Endometrial endothelial cells were more responsive to growth stimulation by VEGF than were dermal endothelial cells isolated under similar conditions. Further characterization indicated that VEGF bound more avidly to HEEC than to other endothelial cell types. CONCLUSIONS: Human endometrial endothelial cells were isolated to homogeneity by a two-part protocol and successfully passaged under culture conditions similar to those used for other endothelial cell types. The HEEC were very responsive to VEGF growth stimulation likely due to elevated affinity, or increased levels of, KDR and FLT 1 on the cell surface. These results indicate that HEEC are capable of maintaining a mature phenotype in culture and might provide a model for understanding the response of these cells to the recurrent cycles of proliferation imposed on the endometrium during menstruation. PMID- 10466116 TI - Confocal laser scanning microscopy of leukocyte adhesion in the microcirculation of the inflamed rat knee joint capsule. AB - OBJECTIVE: The aim of the study was to develop a model of intravital microscopy of the microcirculation of the rat knee joint capsule in acute inflammation with the help of confocal laser scanning microscopy (CLSM). METHODS: The microvascular architecture of the joint capsule was investigated by the use of corrosion casting techniques. Knee joint capsule microcirculation of anesthetized rats could be visualized by dissection of the ligamentum infrapatellare rectum and application of a CLSM. Rhodamine 6G was used as a leukocyte marker. An acute arthritis was induced by intraarticular injection of 0.1 mL of 1% Carrageenan solution. Each experiment lasted for 3 h. Leukocyte adhesion (LA) was measured in 100 microns of vessel length. Leukocyte-rolling velocity (Vwbc), systemic leukocyte count (WBC), and differential blood count (DBC) were monitored in defined intervals. Finally, myeloperoxidase (MPO) activity in frozen-tissue homogenate served as a parameter of neutrophil content. RESULTS: Leukocyte adhesion in carrageenan-treated animals was 9.9 +/- 0.2 leukocytes/100 microns vessel (mean +/- SD; n = 6), while sham- treated animals had 2.8 +/- 0.2. Monoclonal antibodies (MAb) to alpha 4 integrin (0.2 mg/kg) reduced LA to 2.6 +/- 0.1 (p < 0.01). Vwbc/gamma quotients were 0.37 micron s/s in control animals and 0.53 micron s/s in anti-alpha 4-treated animals (p < 0.01). Changes in DBC were marked by lymphopenia and granulocytosis after 180 min. At this time point control animals had 5.1 G/L LC and 2.6 G/I PMN. Animals treated with unspecific antibody had 4.7 G/L LC and 4.9 G/L PMN, while anti-alpha 4-treated animals had 10.6 G/L LC and 2.4 G/L PMN. Photometrically determined extinction of oxidized tetramethylbenzidine (TMB; measure for MPO content) was 1.058 +/- 0.555 in control animals and 0.245 +/- 0.093 in sham-treated animals. Tissue homogenate from unspecific IgG treated group had 0.776 +/- 0.140 and from anti-alpha 4-group 0.334 +/- 0.155 (p < 0.05). CONCLUSIONS: Confocal laser scanning microscopy is a tool for the study of the microcirculation in nontransparent organs. The microcirculation of the acutely inflamed rat knee joint capsule can serve as a model of integrin-mediated LA in vivo. Antiadhesive treatment of animals can reduce the tissue infiltration with inflammatory cells. PMID- 10466117 TI - Nitric oxide inhibits microvascular protein leakage induced by leukocyte adhesion independent and adhesion-dependent inflammatory mediators. AB - OBJECTIVE: The purpose of this study was to determine whether exogenous nitric oxide could block permeability alterations induced by neutrophil-independent (histamine) and neutrophil-dependent (CINC/gro) inflammatory mediators. METHODS: Intravital microscopy was used in the rat mesentery to examine leukocyte adhesion, transvascular FITC-albumin leakage, and perivascular mast cell activation (ruthenium red uptake) in response to local superfusion with histamine or the chemokine CINC/gro. The effects of the nitric oxide donor spermine-NO, or the cGMP analog 8-Br-cGMP were examined. RESULTS: Histamine superfusion increased vascular protein leakage within minutes, but did not increase firm adhesion above that seen in control preparations. The increase in albumin leakage could be prevented by co-administration of spermine-NO, but was not affected by 8-Br-cGMP. CINC/gro elicited a linear increase in vascular protein leakage and a profound increase in leukocyte adhesion. Treatment with spermine-NO or 8-Br-cGMP significantly attenuated increases in both adhesion and albumin leakage. The actions of spermine-NO and 8-Br-cGMP were not due to mast cell stabilization as neither histamine nor CINC/gro elicited mast cell activation. CONCLUSIONS: This study demonstrates that exogenous nitric oxide and 8-Br-cGMP could block adhesion dependent alterations in vascular permeability induced by CINC/gro, while adhesion-independent alterations in permeability induced by histamine could be blocked by exogenous NO but not 8-Br-cGMP. This suggests that different NO dependent signalling pathways are important in modulating these two types of vascular protein leakage. PMID- 10466118 TI - Twelve microsatellite markers for characterization of Plasmodium falciparum from finger-prick blood samples. AB - Multiple, selectively neutral genetic markers are the most appropriate tools for analysis of parasite population structure and epidemiology, but yet existing methods for characterization of malaria field samples utilize a limited number of antigen encoding genes, which appear to be under strong selection. We describe protocols for characterization of 12 microsatellite markers from finger-prick blood samples infected with Plasmodium falciparum. A two-step, heminested strategy was used to amplify all loci, and products were visualized by fluorescent end-labelling of internal primers. This procedure allows amplification from low levels of template, while eliminating the problem of spurious products due to primer carry over from the primary round of PCR. The loci can be conveniently multiplexed, while accurate sizing and quantification of PCR products can be automated using the GENOTYPER software. The primers do not amplify co-infecting malaria species such as P. vivax and P. malariae. To demonstrate the utility of these markers, we characterized 57 infected finger prick blood samples from the village of Mebat in Papua New Guinea for all 12 loci, and all samples were genotyped a second time to measure reproducibility. Numbers of alleles per locus range from 4 to 10 in this population, while heterozygosities range from 0.21 to 0.87. Reproducibility (measured as concordance between predominant alleles detected in replicate samples) ranged from 92 to 98% for the 12 loci. The composition of PCR products from infections containing multiple malaria clones could also be defined using strict criteria and scored in a highly repeatable manner. PMID- 10466119 TI - Anaemia of acute malaria infections in non-immune patients primarily results from destruction of uninfected erythrocytes. AB - While anaemia has long been recognized as a consequence of acute infections with malaria, the relative contributions of direct erythrocyte destruction by parasites, destruction of uninfected erythrocytes and changes in erythropoiesis have been unclear. Fitting of parasitaemia and anaemia data from neurosyphilis patients undergoing malaria therapy to a mathematical model shows that in these patients, an average of 8.5 erythrocytes were destroyed in addition to each erythrocyte observed to become parasitized. The model also showed that dyserythropoiesis plays an insignificant role in the resulting anaemia. The anaemia occurs before a substantial antibody response to parasites or erythrocytes could be generated. We postulate that uninfected erythrocyte destruction occurs through phagocytosis of erythrocytes bound to merozoites killed as a result of the accompanying malaria paroxysms. PMID- 10466120 TI - The relationship of Hammondia hammondi and Sarcocystis mucosa to other heteroxenous cyst-forming coccidia as inferred by phylogenetic analysis of the 18S SSU ribosomal DNA sequence. AB - The complete sequence of the 18S small subunit (SSU) ribosomal DNA of Hammondia hammondi and Sarcocystis mucosa was obtained and compared to SSU rDNA sequences of Neospora caninum, Toxoplasma gondii, Besnoitia besnoiti, 2 species of Frenkelia, 3 species of Isospora, and 13 species of Sarcocystis. Analyses showed that H. hammondi and T. gondii are monophyletic and that these taxa shared a common ancestor with N. caninum and B. besnoiti. The weight of evidence shows that S. mucosa, S. neurona, and Frenkelia species form a clade thereby supporting the conclusion that Sarcocystis is paraphyletic. PMID- 10466121 TI - Successful interrupted feeding of adult Rhipicephalus appendiculatus (Ixodidae) is accompanied by reprogramming of salivary gland protein expression. AB - Ixodid female ticks take one comparatively large bloodmeal which they convert to a single large egg mass and then they die. To examine the outcome of interrupted feeding, equal numbers of male and female Rhipicephalus appendiculatus adult ticks were fed on guinea pigs (host 1) for either 2, 4, or 6 days, or to engorgement (8 days). All of the fully engorged (D8) females laid a single large egg mass (80-160 mg/tick), while 85% of the day 6-fed (D6) female ticks (n = 20) each laid a small egg mass (6.1 mg/tick). None of the females that had fed for 2 or 4 days oviposited. Ninety percent (n = 20) of the day 2-fed (D2) females survived for 4 weeks after their feeding was interrupted, whereas 65% (n = 20) of the day 4-fed (D4) females survived. All of the surviving partially fed female ticks (D2 and D4) attached to a second guinea pig (host 2) and attained engorged body weights that were not significantly different from those of the control females (P < 0.05). Female ticks that engorged following interrupted feeding layed egg masses comparable to the controls, indicating that engorgement on host 2 was successful. The salivary gland protein profile of female ticks changed constantly during feeding. However, when feeding was interrupted, the protein expression pattern switched back to that of the non-parasitic state, presumably to enable the partially fed ticks to survive and reattach on the new host. This observation indicates that female ixodid ticks have a natural ability to survive and re-establish successful feeding on a new host if the first attempt at feeding is unsuccessful. Such an interrupted feeding mechanism supports the hypothesis that partially engorged ticks may play a role in tick-borne pathogen transmission. PMID- 10466122 TI - The effect of pregnancy on Wuchereria bancrofti microfilarial load in humans. AB - As part of a drug trial against bancroftian filariasis in the East Sepik Province of Papua New Guinea we measured the pre-treatment microfilarial densities of 2219 individuals. Mean levels generally increased with age in both sexes, with a tendency to plateau at the highest ages. However, there was a reduction among women of approximately reproductive age. Allowing for the tendency for aggregation to decrease with age, this reduction was statistically significant. However, a comparison of pregnant women and controls showed no evidence that the reduction is specifically related to pregnancy. Moreover, a simple differential equation model of microfilarial acquisition and loss suggests that age-specific patterns of exposure are also unlikely to be solely responsible. Therefore, we suggest that the observed reduction in microfilarial intensity may result from hormonal changes associated with female reproduction, possibly in combination with other factors. PMID- 10466123 TI - Protease activity in the larval stage of the parasitoid wasp, Eulophus pennicornis (Nees) (Hymenoptera: Eulophidae); effects of protease inhibitors. AB - Hymenopteran, parasitoid wasps have good potential for use in integrated pest management (IPM); for example, the gregarious ectoparasitoid, Eulophus pennicornis, has been suggested as a biological control agent for larvae of the tomato moth (Lacanobia oleracea L.). However, the processes by which such parasitic larvae are able to utilize the nutritional resource provided by the host have been little studied. Protease activity was present in E. pennicornis larvae, and characterization of the enzymes responsible for proteolysis was performed using a range of synthetic substrates and specific inhibitors. Serine protease enzymes was both trypsin- and chymotrypsin-like activities were present. A range of plant-derived serine protease inhibitors was tested for activity against these enzymes. Certain inhibitors, notably soybean Kunitz inhibitor (SKTI), inhibited enzyme activity by > 80% at < 10(-5) M. When SKTI was fed to L. oleracea larvae in an artificial diet, the inhibitor was subsequently detected within the larval haemolymph, showing that protease inhibitors in the host diet can be delivered to a parasitoid via the host haemolymph. If transgenic plants expressing foreign protease inhibitors for protection against insect pests are to form a component of IPM systems, possible adverse effects, whether direct or indirect, of transgene expression on parasitoids like E. pennicornis should be considered. PMID- 10466124 TI - Direct analysis of the secretions of the potato cyst nematode Globodera rostochiensis. AB - Secretions were induced from second (invasive) stage juveniles (J2s) of the potato cyst nematode Globodera rostochiensis by exposing them to 5-methoxy-N,N dimethyl tryptamine oxalate (DMT). Secretions were collected from J2s in sufficient quantity to allow direct analysis. Gel electrophoresis followed by monochromatic silver staining demonstrated the presence of at least 10 proteins. The presence of several enzymes, including superoxide dismutase and proteases, was demonstrated using Western blots and activity assays. Antisera raised against the secretions recognized bands on Western blots consistent in molecular mass with those identified on silver stained gels. The antisera recognized structures implicated in the production of secretions including the subventral gland cells and surface of J2s. PMID- 10466125 TI - The structure of the monogenean community on the gills of Pimelodus maculatus in Rio de la Plata (Argentina). AB - The Monogenea fauna was studied from the gills of the catfish Pimelodus maculatus. Fish were caught from Rio de la Plata, in Buenos Aires harbour (Argentina). Five species of Monogenea were found. The structure of the monogenean community was analysed at the component and infracommunity level. The nature of the monogenean community, its possible interactivity, seasonal variation and predictability are also discussed. PMID- 10466126 TI - Niche preferences and spatial distribution of Monogenea on the gills of Pimelodus maculatus in Rio de la Plata (Argentina). AB - Five Monogenea species were found on the gills of the catfish Pimelodus maculatus in Rio de la Plata (Argentina). These were used for studying the preference of species on different gill-hemibranches, niche breadth and niche overlap between species. It was found that congeneric species had a generic-specific preference for certain gill-hemibranches. Niche breadth appeared to be related to the number of individuals of each species. Niche overlap between the species is discussed. PMID- 10466127 TI - The isolation of differentially expressed cDNA clones from the filarial nematode Brugia pahangi. AB - A cDNA library constructed from 3 day post-infective L3 of the filarial nematode Brugia pahangi was screened by differential hybridization with cDNA probes prepared from different life-cycle stages. Five cDNA clones hybridizing selectively to the mosquito-derived L3 probe were isolated and characterized. Northern blot analysis of 4 of the clones confirmed that each was most highly expressed in the mosquito-derived L3. The expression of each mRNA during parasite development in the mosquito vector was investigated using RT-PCR, and all were shown to be abundant in the immature L3. Four of the 5 cDNAs cloned coded for structural proteins: 2 cuticular collagens, and the muscle proteins tropomyosin and troponin. Further studies on troponin using an antiserum raised to the recombinant protein demonstrated that the protein, unlike the mRNA, was present in all life-cycle stages examined, while immunogold labelling demonstrated that it was localized to the muscle blocks. PMID- 10466128 TI - Enhancement of apoptosis with loss of cellular adherence in the villus epithelium of the small intestine after infection with the nematode Nippostrongylus brasiliensis in rats. AB - It has been reported that infection with Nippostrongylus brasiliensis induces villus atrophy with various histological alterations. In N. brasiliensis-infected rats, villus length in the jejunum was reduced significantly at day 10 p.i., when serum levels of rat mast cell protease (RMCP) II had increased significantly. To determine whether the villus atrophy is associated with enhancement of apoptosis, apoptotic nuclei were labelled using the nick end-labelling method. Numbers of labelled cells were markedly increased in the villus epithelium at 7-10 days p.i., while the numbers returned to normal 14 days p.i. when worms were rejected from the intestine and villus length became normal. Examination of the expression of the adhesion molecule E-cadherin showed granular immunoreactivity in the cytoplasm of atrophic villus epithelium with loss of normal localization to epithelial cell borders. In mast cell-deficient Ws/Ws rats, villus length was reduced as significantly as in +/+ counterparts at day 10 p.i. with marked increases in the numbers of apoptotic cells. These results suggested that villus atrophy was closely associated with enhanced apoptosis and loss of adhesion in epithelial cells. Mast cell activation appears not to be involved in these alterations. PMID- 10466129 TI - Seasonal variation in development and survival of Ascaris suum and Trichuris suis eggs on pastures. AB - Pig faeces were deposited on experimental plots in the spring, summer, autumn and winter to study development and survival of Ascaris suum and Trichuris suis eggs under outdoor conditions. Faeces were placed either in short grass or 2 cm below the surface of bare soil, imitating pastures used by nose-ringed, grazing pigs or normally rooting pigs, respectively. The numbers and developmental stages of the eggs were recorded in faeces and soil for up to 50 weeks post-deposition. Embryonation took place only during the summer months and seemingly was independent of the microclimate. The majority of A. suum and T. suis eggs, which are generally considered to be extremely resistant and long-lived, seems to disappear rather fast. The disappearance rate for A. suum eggs was higher than for T. suis eggs, and both egg types disappeared significantly faster in the summer months than in the winter months, and when placed in short grass than when buried in soil (less exposed). We discuss how knowledge on egg development and survival may be used in the planning of pasture strategies for control of helminth infections in outdoor pigs. PMID- 10466130 TI - A second peak of egg excretion in Strongyloides ratti-infected rats: its origin and biological meaning. AB - In Strongyloides ratti-infected rats, 2 peaks of egg excretion were observed; a large one with maximum egg production on days 7-8 of infection and a small more inconspicuous one around day 25. The second peak, which had been ignored in most studies, was produced by adults in the caecum and the colon. The adults were larger in length and had more embryonated eggs in the uterus compared with adults in the small intestine at day 25 post-infection. It is suggested that parasitic adults once expelled from the small intestine resettle and recover in the large intestine. Filter paper faecal culture carried out for 9 days at different days post-infection revealed that the total number of infective larvae that developed during the second peak was twice the number that developed during the first peak, despite the fact that total egg output during the second peak was less than one twentieth of the first peak. The results suggest that the small second peak was as important as the first one in the transmission of S. ratti. PMID- 10466131 TI - Heterogeneity in the distribution of Strongyloides ratti infective stages among the faecal pellets of rats. AB - The distribution of helminth parasites within their host population is usually overdispersed and can be described by the negative binomial distribution. The causes of this overdispersion are poorly understood, but heterogeneity in the distribution of infective stages within the environment has been implicated as a possible factor. Here we describe the distribution of infective stages of the rat intestinal nematode parasite Strongyloides ratti among the faecal pellets of its host. The distribution of infective stages between faecal pellets is overdispersed and well described by the negative binomial distribution. This overdispersion increases during the course of infection and occurs over a range of infection intensities. Overdispersion of nematode infective stages among faecal pellets may result in increased spatial heterogeneity of the infective stages in the environment and thus may contribute to the generation of overdispersion of adult parasitic stages. In addition, these findings raise important issues regarding the accurate quantification of helminth egg counts from faecal samples. PMID- 10466132 TI - The utility of complete genome sequences in the study of pathogenic bacteria. AB - The availability of complete genome sequences is a revolution in the study of microorganisms. A fully annotated genome sequence provides an interactive tool for scientists and influences the approach and focus of research. In this article I discuss the impact of genome sequencing projects of bacteria. Much useful data have been obtained but the experimental methods needed to fully exploit the information continue to develop. Some of the approaches and particular applications relevant to bacteria of clinical importance are discussed. PMID- 10466133 TI - Status of protozoan genome analysis: trypanosomatids. AB - The three trypanosomatid genome projects have employed common strategies which include: analysis of pulsed-field gel electrophoretic chromosomal karyotypes; physical mapping using big DNA (cosmid, pacmid P1, bacterial artificial chromosome, yeast artificial chromosome) libraries; partial cDNA sequence analysis to develop sets of expressed sequence tags (ESTs) for gene discovery and use as markers in physical mapping; genomic sequencing; dissemination of information through development of web-sites and ACeDB-based fully integrated databases; and establishment of functional genomics programmes to maximize useful application of genome data. Highlights of the projects to date have been the demonstration that, despite extensive chromosomal size polymorphisms for diploid homologues within Africa trypanosomes, T. cruzi or Leishmania, the physical linkage groups for markers on each chromosome are retained across all isolates/species studied within each group. For African trypanosomes, detailed analysis of chromosome 1 has demonstrated that repetitive sequences and the two retroposon-like elements RIME and INGI are localized to a defined region at one end of the chromosome, with the bulk of the central region of the chromosome containing genes coding for expressed proteins. Comparative mapping shows that, although subtelomeric changes account for a large proportion of the polymorphism in chromosome size in African trypanosomes, there are significant expansions and contractions in regions across the entire chromosome. The highlight of the genomic sequencing projects has been the demonstration of just 2 putative transcriptional units of chromosome 1 of Leishmania major, extending on opposite strands from a point in the central region of the chromosome. A similar observation made on 93.4 kb of contiguous sequence for T. cruzi chromosome 3 suggests the presence of promoter and regulatory elements at the junctions of large polycistronic transcriptional units. All data obtained from the genome projects are made available through the public domain, which has prompted changing philosophies in how we approach analysis of the biology of these organisms, and strategies that we can employ now in the search for new therapies and vaccines. PMID- 10466134 TI - Data mining parasite genomes: haystack searching with a computer. AB - A number of genomes of parasitic organisms are presently being sequenced in the public domain, including Plasmodium falciparum, Leishmania major and Trypanosoma brucei with the likelihood of at least expressed sequence tag (EST) projects for several filarial and apicomplexan species. The early and timely release of sequence data to the community via the World Wide Web (www), and the public databases, (EMBL and GENBANK), forms an invaluable resource. Data mining, or 'haystack searching' this resource is becoming more fruitful to all members of the scientific community as the volume of data, diversity of genomes sampled, and accessibility increase. PMID- 10466135 TI - Helminth genome analysis: the current status of the filarial and schistosome genome projects. Filarial Genome Project. Schistosome Genome Project. AB - Genome projects for the parasitic helminths Brugia malayi (a representative filarial nematode) and Schistosoma were initiated in 1995 by the World Health Organization with the ultimate objectives of identifying new vaccine candidates and drug targets and of developing low resolution genome maps. Because no genetic maps are available, and very few genes have been characterized from either parasite group, the first goal of both Initiatives has been to catalogue new genes for future placement on chromosome and physical maps. These genes have been identified by the expressed sequence tag (EST) approach, utilising cDNA libraries constructed from diverse life cycle stages. To date, the Initiatives have deposited over 16,000 Brugia ESTs and nearly 8000 Schistosoma ESTs in Genbank's dbEST database, corresponding to 6000 and over 3600 genes respectively (33% of Brugia's estimated gene compliment, 18-24% of that of Schistosoma). Large fragment, genomic libraries have been constructed in BAC and YAC vectors for studies of genomic organization and for physical and chromosome mapping, and public, hypertext genomic databases have been established to facilitate data access. We present a summary of progress within the helminth genome initiatives and give several examples of important gene discoveries and future applications of these data. PMID- 10466136 TI - Parasitic helminth genomics. Filarial Genome Project. AB - The initiation of genome projects on helminths of medical importance promises to yield new drug targets and vaccine candidates in unprecedented numbers. In order to exploit this emerging data it is essential that the user community is aware of the scope and quality of data available, and that the genome projects provide analyses of the raw data to highlight potential genes of interest. Core bioinformatics support for the parasite genome projects has promoted these approaches. In the Brugia genome project, a combination of expressed sequence tag sequencing from multiple cDNA libraries representing the complete filarial nematode lifecycle, and comparative analysis of the sequence dataset, particularly using the complete genome sequence of the model nematode C. elegans, has proved very effective in gene discovery. PMID- 10466137 TI - The importance of reverse genetics in determining gene function in apicomplexan parasites. AB - The phylum Apixomplexa includes obligate intracellular parasites that are of enormous medical and veterinary significance, as they are responsible for a wide variety of diseases including malaria, toxoplasmosis, coccidiosis, cryptosporidiosis, theileriosis and babesiosis. The EST sequencing projects in Toxoplasma gondii and the Plasmodium falciparum genome sequencing project have greatly accelerated gene discovery, revealing for example novel coding sequences restricted to the Apicomplexa. However, easy acquisition of sequence is almost useless if the function of any given gene cannot be tested. The establishment of transfection systems in Toxoplasma gondii, Neospora and in several Plasmodium species has provided us with the reverse genetics methods appropriate to the functional analysis of genes. Over the past few years, the discovery of novel genes coupled to the ability to introduce or modify genes has already contributed to a better understanding of cell biology and pathogenesis of these obligate intracellular parasites. Some insights into the complex processes of parasite invasion, differentiation, regulation of gene expression and protein trafficking are emerging although identification of the exact functional roles for many molecules is still awaiting more investigation. This review summarizes progress in this area. It also emphasises the tight link and synergy between Toxoplasma and malaria research. The use of reverse genetics does not guarantee the answer to gene function, so we can learn from both failed and successful experiments about how better and more efficiently to use 'genomics' to accelerate discoveries relevant to the understanding of parasitism by Apicomplexa. PMID- 10466138 TI - Characterization of plant nematode genes: identifying targets for a transgenic defence. AB - Current control of plant parasitic nematodes often relies on highly toxic and environmentally harmful nematicides. As their use becomes increasingly restricted there is an urgent need to develop crop varieties with resistance to nematodes. The limitations surrounding conventional plant breeding ensure there is a clear opportunity for transgenic resistance to lessen current dependence on chemical control. The increasing use of molecular biology techniques in the field of plant nematology is now providing useful information for the design of novel defences to meet the new needs. Plant responses to parasitism are being investigated at the molecular level and nematode gene products that could be targets for a direct anti-nematode defence are being characterized. The potential of an anti-feedant approach to nematode control has been demonstrated. It is based on the transgenic expression of proteinase inhibitors. The rational development of this strategy involves characterization of nematode proteinase genes and optimization of inhibitors by protein engineering. Durability of the resistance can be enhanced by stacking transgenes directed at different nematode targets. PMID- 10466139 TI - Whole genome genetic-typing in yeast using high-density oligonucleotide arrays. AB - Genome sequence information in combination with new technologies has allowed researchers to approach genetic problems in new ways. High-density oligonucleotide arrays were used to probe the genome content of the yeast Saccharomyces cerevisiae. We show that these arrays, containing oligonucleotides complementary to the sequenced strain of S. cerevisiae, can be used to identify open reading frames that are missing or present in higher or lower copy number in related isolates of S. cerevisiae. We apply this method to the characterization of the genome of a strain derived from a clinical isolate of S. cerevisiae. Our results show that the telomeres are the regions with the most variability between the two strains. PMID- 10466140 TI - Electroencephalographic imaging of higher brain function. AB - High temporal resolution is necessary to resolve the rapidly changing patterns of brain activity that underlie mental function. Electroencephalography (EEG) provides temporal resolution in the millisecond range. However, traditional EEG technology and practice provide insufficient spatial detail to identify relationships between brain electrical events and structures and functions visualized by magnetic resonance imaging or positron emission tomography. Recent advances help to overcome this problem by recording EEGs from more electrodes, by registering EEG data with anatomical images, and by correcting the distortion caused by volume conduction of EEG signals through the skull and scalp. In addition, statistical measurements of sub-second interdependences between EEG time-series recorded from different locations can help to generate hypotheses about the instantaneous functional networks that form between different cortical regions during perception, thought and action. Example applications are presented from studies of language, attention and working memory. Along with its unique ability to monitor brain function as people perform everyday activities in the real world, these advances make modern EEG an invaluable complement to other functional neuroimaging modalities. PMID- 10466141 TI - Functionally independent components of early event-related potentials in a visual spatial attention task. AB - Spatial visual attention modulates the first negative-going deflection in the human averaged event-related potential (ERP) in response to visual target and non target stimuli (the N1 complex). Here we demonstrate a decomposition of N1 into functionally independent subcomponents with functionally distinct relations to task and stimulus conditions. ERPs were collected from 20 subjects in response to visual target and non-target stimuli presented at five attended and non-attended screen locations. Independent component analysis, a new method for blind source separation, was trained simultaneously on 500 ms grand average responses from all 25 stimulus-attention conditions and decomposed the non-target N1 complexes into five spatially fixed, temporally independent and physiologically plausible components. Activity of an early, laterally symmetrical component pair (N1aR and N1aL) was evoked by the left and right visual field stimuli, respectively. Component N1aR peaked ca. 9 ms earlier than N1aL. Central stimuli evoked both components with the same peak latency difference, producing a bilateral scalp distribution. The amplitudes of these components were no reliably augmented by spatial attention. Stimuli in the right visual field evoked activity in a spatio temporally overlapping bilateral component (N1b) that peaked at ca. 180 ms and was strongly enhanced by attention. Stimuli presented at unattended locations evoked a fourth component (P2a) peaking near 240 ms. A fifth component (P3f) was evoked only by targets presented in either visual field. The distinct response patterns of these components across the array of stimulus and attention conditions suggest that they reflect activity in functionally independent brain systems involved in processing attended and unattended visuospatial events. PMID- 10466142 TI - Magnetoencephalography in the study of human somatosensory cortical processing. AB - Magnetoencephalography (MEG) is a totally non-invasive research method which provides information about cortical dynamics on a millisecond time-scale. Whole scalp magnetic field patterns following stimulation of different peripheral nerves indicate activation of an extensive cortical network. At the SI cortex, the responses reflect mainly the activity of area 3b, with clearly somatotopical representations of different body parts. The SII cortex is activated bilaterally and it also receives, besides tactile input, nociceptive afference. Somatically evoked MEG signals may also be detected from the posterior parietal cortex, central mesial cortex and the frontal lobe. The serial versus parallel processing in the cortical somatosensory network is still under debate. PMID- 10466143 TI - Cellular mechanisms of brain energy metabolism and their relevance to functional brain imaging. AB - Despite striking advances in functional brain imaging, the cellular and molecular mechanisms that underlie the signals detected by these techniques are still largely unknown. The basic physiological principle of functional imaging is represented by the tight coupling existing between neuronal activity and the associated local increase in both blood flow and energy metabolism. Positron emission tomography (PET) signals detect blood flow, oxygen consumption and glucose use associated with neuronal activity; the degree of blood oxygenation is currently thought to contribute to the signal detected with functional magnetic resonance imaging, while magnetic resonance spectroscopy (MRS) identifies the spatio-temporal pattern of the activity-dependent appearance of certain metabolic intermediates such as glucose or lactate. Recent studies, including those of neurotransmitter-regulated metabolic fluxes in purified preparations and analyses of the cellular localization of enzymes and transporters involved in energy metabolism, as well as in vivo microdialysis and MRS approaches have identified the neurotransmitter glutamate and astrocytes, a specific type of glial cell, as pivotal elements in the coupling of synaptic activity with energy metabolism. Astrocytes are ideally positioned to sense increases in synaptic activity and to couple them with energy metabolism. Indeed they possess specialized processes that cover the surface of intraparenchymal capillaries, suggesting that astrocytes may be a likely site of prevalent glucose uptake. Other astrocyte processes are wrapped around synaptic contacts which possess receptors and reuptake sites for neurotransmitters. Glutamate stimulates glucose uptake into astrocytes. This effect is mediated by specific glutamate transporters present on these cells. The activity of these transporters, which is tightly coupled to the synaptic release of glutamate and operates the clearance of glutamate from the extracellular space, is driven by the electrochemical gradient of Na+. This Na(+) dependent uptake of glutamate into astrocytes triggers a cascade of molecular events involving the Na+/K(+)-ATPase leading to the glycolytic processing of glucose and the release of lactate by astrocytes. The stoichiometry of this process is such that for one glutamate molecule taken up with three Na+ ions, one glucose molecule enters an astrocyte, two ATP molecules are produced through aerobic glycolysis and two lactate molecules are released. Within the astrocyte, one ATP molecule fuels one 'turn of the pump' while the other provides the energy needed to convert glutamate to glutamine by glutamine synthase. Evidence has been accumulated from structural as well as functional studies indicating that, under aerobic conditions, lactate may be the preferred energy substrate of activated neurons. Indeed, in the presence of oxygen, lactate is converted to pyruvate, which can be processed through the tricarboxylic acid cycle and the associated oxidative phosphorylation, to yield 17 ATP molecules per lactate molecule. These data suggest that during activation the brain may transiently resort to aerobic glycolysis occurring in astrocytes, followed by the oxidation of lactate by neurons. The proposed model provides a direct mechanism to couple synaptic activity with glucose use and is consistent with the notion that the signals detected during physiological activation with 18F-deoxyglucose (DG)-PET may reflect predominantly uptake of the tracer into astrocytes. This conclusion does not question the validity of the 2-DG-based techniques, rather it provides a cellular and molecular basis for these functional brain imaging techniques. PMID- 10466144 TI - In vivo nuclear magnetic resonance spectroscopy studies of the relationship between the glutamate-glutamine neurotransmitter cycle and functional neuroenergetics. AB - In this article we review recent studies, primarily from our laboratory, using 13C NMR (nuclear magnetic resonance) to non-invasively measure the rate of the glutamate-glutamine neurotransmitter cycle in the cortex of rats and humans. In the glutamate-glutamine cycle, glutamate released from nerve terminals is taken up by surrounding glial cells and returned to the nerve terminals as glutamine. 13C NMR studies have shown that the rate of the glutamate-glutamine cycle is extremely high in both the rat and human cortex, and that it increases with brain activity in an approximately 1:1 molar ratio with oxidative glucose metabolism. The measured ratio, in combination with proposals based on isolated cell studies by P. J. Magistretti and co-workers, has led to the development of a model in which the majority of brain glucose oxidation is mechanistically coupled to the glutamate-glutamine cycle. This model provides the first testable mechanistic relationship between cortical glucose metabolism and a specific neuronal activity. We review here the experimental evidence for this model as well as implications for blood oxygenation level dependent magnetic resonance imaging and positron emission tomography functional imaging studies of brain function. PMID- 10466145 TI - Functional magnetic resonance imaging: imaging techniques and contrast mechanisms. AB - Functional magnetic resonance imaging (fMRI) is a widely used technique for generating images or maps of human brain activity. The applications of the technique are widespread in cognitive neuroscience and it is hoped they will eventually extend into clinical practice. The activation signal measured with fMRI is predicated on indirectly measuring changes in the concentration of deoxyhaemoglobin which arise from an increase in blood oxygenation in the vicinity of neuronal firing. The exact mechanisms of this blood oxygenation level dependent (BOLD) contrast are highly complex. The signal measured is dependent on both the underlying physiological events and the imaging physics. BOLD contrast, although sensitive, is not a quantifiable measure of neuronal activity. A number of different imaging techniques and parameters can be used for fMRI, the choice of which depends on the particular requirements of each functional imaging experiment. The high-speed MRI technique, echo-planar imaging provides the basis for most fMRI experiments. The problems inherent to this method and the ways in which these may be overcome are particularly important in the move towards performing functional studies on higher field MRI systems. Future developments in techniques and hardware are also likely to enhance the measurement of brain activity using MRI. PMID- 10466146 TI - Functional mapping in the human brain using high magnetic fields. AB - An avidly pursued new dimension in magnetic resonance imaging (MRI) research is the acquisition of physiological and biochemical information non-invasively using the nuclear spins of the water molecules in the human body. In this trial, a recent and unique accomplishment was the introduction of the ability to map human brain function non-invasively. Today, functional images with subcentimetre resolution of the entire human brain can be generated in single subjects and in data acquisition times of several minutes using 1.5 tesla (T) MRI scanners that are often used in hospitals for clinical purposes. However, there have been accomplishments beyond this type of imaging using significantly higher magnetic fields such as 4 T. Efforts for developing high magnetic field human brain imaging and functional mapping using MRI (fMRI) were undertaken at about the same time. It has been demonstrated that high magnetic fields result in improved contrast and, more importantly, in elevated sensitivity to capillary level changes coupled to neuronal activity in the blood oxygenation level dependent (BOLD) contrast mechanism used in fMRI. These advantages have been used to generate, for example, high resolution functional maps of ocular dominance columns, retinotopy within the small lateral geniculate nucleus, true single trial fMRI and early negative signal changes in the temporal evolution of the BOLD signal. So far these have not been duplicated or have been observed as significantly weaker effects at much lower field strengths. Some of these high field advantages and accomplishments are reviewed in this paper. PMID- 10466147 TI - Event-related functional magnetic resonance imaging: modelling, inference and optimization. AB - Event-related functional magnetic resonance imaging is a recent and popular technique for detecting haemodynamic responses to brief stimuli or events. However, the design of event-related experiments requires careful consideration of numerous issues of measurement, modelling and inference. Here we review these issues, with particular emphasis on the use of basis functions within a general linear modelling framework to model and make inferences about the haemodynamic response. With these models in mind, we then consider how the properties of functional magnetic resonance imaging data determine the optimal experimental design for a specific hypothesis, in terms of stimulus ordering and interstimulus interval. Finally, we illustrate various event-related models with examples from recent studies. PMID- 10466148 TI - Transcranial magnetic stimulation: studying the brain-behaviour relationship by induction of 'virtual lesions'. AB - Transcranial magnetic stimulation (TMS) provides a non-invasive method of induction of a focal current in the brain and transient modulation of the function of the targeted cortex. Despite limited understanding about focality and mechanisms of action, TMS provides a unique opportunity of studying brain behaviour relations in normal humans. TMS can enhance the results of other neuroimaging techniques by establishing the causal link between brain activity and task performance, and by exploring functional brain connectivity. PMID- 10466149 TI - Statistical limitations in functional neuroimaging. I. Non-inferential methods and statistical models. AB - Functional neuroimaging (FNI) provides experimental access to the intact living brain making it possible to study higher cognitive functions in humans. In this review and in a companion paper in this issue, we discuss some common methods used to analyse FNI data. The emphasis in both papers is on assumptions and limitations of the methods reviewed. There are several methods available to analyse FNI data indicating that none is optimal for all purposes. In order to make optimal use of the methods available it is important to know the limits of applicability. For the interpretation of FNI results it is also important to take into account the assumptions, approximations and inherent limitations of the methods used. This paper gives a brief overview over some non-inferential descriptive methods and common statistical models used in FNI. Issues relating to the complex problem of model selection are discussed. In general, proper model selection is a necessary prerequisite for the validity of the subsequent statistical inference. The non-inferential section describes methods that, combined with inspection of parameter estimates and other simple measures, can aid in the process of model selection and verification of assumptions. The section on statistical models covers approaches to global normalization and some aspects of univariate, multivariate, and Bayesian models. Finally, approaches to functional connectivity and effective connectivity are discussed. In the companion paper we review issues related to signal detection and statistical inference. PMID- 10466151 TI - If neuroimaging is the answer, what is the question? AB - It is unclear that we will come to a better understanding of mental processes simply by observing which neural loci are activated while subjects perform a task. Rather, I suggest here that it is better to come armed with a question that directs one to design tasks in ways that take advantage of the strengths of neuroimaging techniques (particularly positron emission tomography and functional magnetic resonance imaging). Here I develop a taxonomy of types of questions that can be easily addressed by such techniques. The first class of questions focuses on how information processing is implemented in the brain; these questions can be posed at a very coarse scale, focusing on the entire system that confers a particular ability, or at increasingly more specific scales, ultimately focusing on individual structures or processes. The second class of questions focuses on specifying when particular processes and structures are invoked; these questions focus on how one can use patterns of activation to infer that specific processes and structures were invoked, and on how processing changes in different circumstances. The use of neuroimaging to address these questions is illustrated with results from experiments on visual cognition, and caveats regarding the logic of inference in each case are noted. Finally, the necessary interplay between neuroimaging and behavioural studies is stressed. PMID- 10466150 TI - Statistical limitations in functional neuroimaging. II. Signal detection and statistical inference. AB - The field of functional neuroimaging (FNI) methodology has developed into a mature but evolving area of knowledge and its applications have been extensive. A general problem in the analysis of FNI data is finding a signal embedded in noise. This is sometimes called signal detection. Signal detection theory focuses in general on issues relating to the optimization of conditions for separating the signal from noise. When methods from probability theory and mathematical statistics are directly applied in this procedure it is also called statistical inference. In this paper we briefly discuss some aspects of signal detection theory relevant to FNI and, in addition, some common approaches to statistical inference used in FNI. Low-pass filtering in relation to functional-anatomical variability and some effects of filtering on signal detection of interest to FNI are discussed. Also, some general aspects of hypothesis testing and statistical inference are discussed. This includes the need for characterizing the signal in data when the null hypothesis is rejected, the problem of multiple comparisons that is central to FNI data analysis, omnibus tests and some issues related to statistical power in the context of FNI. In turn, random field, scale space, non parametric and Monte Carlo approaches are reviewed, representing the most common approaches to statistical inference used in FNI. Complementary to these issues an overview and discussion of non-inferential descriptive methods, common statistical models and the problem of model selection is given in a companion paper. In general, model selection is an important prelude to subsequent statistical inference. The emphasis in both papers is on the assumptions and inherent limitations of the methods presented. Most of the methods described here generally serve their purposes well when the inherent assumptions and limitations are taken into account. Significant differences in results between different methods are most apparent in extreme parameter ranges, for example at low effective degrees of freedom or at small spatial autocorrelation. In such situations or in situations when assumptions and approximations are seriously violated it is of central importance to choose the most suitable method in order to obtain valid results. PMID- 10466152 TI - The challenge of non-invasive cognitive physiology of the human brain: how to negotiate the irrelevant background noise without spoiling the recorded data through electronic averaging. AB - Brain mechanisms involved in selective attention in humans can be studied by measures of regional blood flow and metabolism (by positron emission tomography) which help identify the various locations with enhanced activities over a period of time of seconds. The physiological measures provided by scalp-recorded brain electrical potentials have a better resolution (milliseconds) and can reveal the actual sequences of distinct neural events and their precise timing. We studied selective attention to sensory inputs from fingers because the brain somatic representations are deployed over the brain convexity under the scalp thereby making it possible to assess distinct stages of cortical processing and representation through their characteristic scalp topographies. In the electrical response to a finger input attended by the subject, the well-known P300 manifests a widespread inhibitory mechanism which is released after a target stimulus has been identified. P300 is preceded by distinct cognitive electrogeneses such as P40, P100 and N140 which can be differentiated from the control (obligatory) profile by superimposition or electronic subtraction. The first cortical response N20 is stable across conditions, suggesting that the first afferent thalamocortical volley is not affected by selective attention. At the next stage of modality-specific cortex in which the sensory features are processed and represented, responses were enhanced (cognitive P40) only a very few milliseconds after arrival of the afferent volley at the cortex, thus documenting a remarkable precocity of attention gain control in the somatic modality. The physiology of selective attention also provides useful cues in relation to non-target inputs which the subject must differentiate in order to perform the task. When having to tell fingers apart, the brain strategy for non-target fingers is not to inhibit or filter them out, but rather to submit their input to several processing operations that are actually enhanced when the discrimination from targets becomes more difficult. While resolving a number of such issues, averaged data cannot disclose the flexibility of brain mechanisms nor the detailed features of cognitive electrogeneses because response variations along time have been ironed out by the bulk treatment. We attempted to address the remarkable versatility of humans in dealing with their sensory environment under ecological conditions by studying single non-averaged responses. We identified distinct cognitive P40, P100, N140 and P300 electrogeneses in spite of the noise by numerically assessing their characteristic scalp topography signatures. Single-trial data suggest reconsiderations of current psychophysiological issues. The study of non-averaged responses can clarify issues raised by averaging studies as illustrated by our recent study of cognitive brain potentials for finger stimuli which remain outside the subject's awareness. This has to do with the physiological basis of the 'cognitive unconscious', that is, current mental processes lying on the fringe or outside of phenomenal awareness and voluntary control, but which can influence ongoing behaviour. Averaged data suggest that, in selective auditory attention, the subject may not notice mild concomitant finger inputs. The study of non-averaged responses documents the optional and independent occurrence of the cognitive P40, P100 and N140 (but not P300) electrogeneses while the finger inputs remain outside phenomenal awareness. These results suggest that the subject unconsciously assigns limited cognitive resources to distinct somatic cortical areas thereby submitting finger inputs to an intermittent curtailed surveillance which can remain on the fringe or outside consciousness. The study of cognitive electrogeneses in single non-averaged responses is making possible a neurophysiology of cognition in real time. PMID- 10466153 TI - When encoding yields remembering: insights from event-related neuroimaging. AB - To understand human memory, it is important to determine why some experiences are remembered whereas others are forgotten. Until recently, insights into the neural bases of human memory encoding, the processes by which information is transformed into an enduring memory trace, have primarily been derived from neuropsychological studies of humans with select brain lesions. The advent of functional neuroimaging methods, such as positron emission tomography (PET) and functional magnetic resonance imaging (fMRI), has provided a new opportunity to gain additional understanding of how the brain supports memory formation. Importantly, the recent development of event-related fMRI methods now allows for examination of trial-by-trial differences in neural activity during encoding and of the consequences of these differences for later remembering. In this review, we consider the contributions of PET and fMRI studies to the understanding of memory encoding, placing a particular emphasis on recent event-related fMRI studies of the Dm effect: that is, differences in neural activity during encoding that are related to differences in subsequent memory. We then turn our attention to the rich literature on the Dm effect that has emerged from studies using event related potentials (ERPs). It is hoped that the integration of findings from ERP studies, which offer higher temporal resolution, with those from event-related fMRI studies, which offer higher spatial resolution, will shed new light on when and why encoding yields subsequent remembering. PMID- 10466155 TI - Pain and functional imaging. AB - Functional neuroimaging has fundamentally changed our knowledge about the cerebral representation of pain. For the first time it has been possible to delineate the functional anatomy of different aspects of pain in the medial and lateral pain systems in the brain. The rapid developments in imaging methods over the past years have led to a consensus in the description of the central pain responses between different studies and also to a definition of a central pain matrix with specialized subfunctions in man. In the near future we will see studies where a systems perspective allows for a better understanding of the regulatory mechanisms in the higher-order frontal and parietal cortices. Also, pending the development of experimental paradigms, the functional anatomy of the emotional aspects of pain will become better known. PMID- 10466154 TI - Spatial attention and neglect: parietal, frontal and cingulate contributions to the mental representation and attentional targeting of salient extrapersonal events. AB - The syndrome of contralesional neglect reflects a lateralized disruption of spatial attention. In the human, the left hemisphere shifts attention predominantly in the contralateral hemispace and in a contraversive direction whereas the right hemisphere distributes attention more evenly, in both hemispaces and both directions. As a consequence of this asymmetry, severe contralesional neglect occurs almost exclusively after right hemisphere lesions. Patients with left neglect experience a loss of salience in the mental representation and conscious perception of the left side and display a reluctance to direct orientating and exploratory behaviours to the left. Neglect is distributed according to egocentric, allocentric, world-centred, and object centred frames of reference. Neglected events can continue to exert an implicit influence on behaviour, indicating that the attentional filtering occurs at the level of an internalized representation rather than at the level of peripheral sensory input. The unilateral neglect syndrome is caused by a dysfunction of a large-scale neurocognitive network, the cortical epicentres of which are located in posterior parietal cortex, the frontal eye fields, and the cingulate gyrus. This network coordinates all aspects of spatial attention, regardless of the modality of input or output. It helps to compile a mental representation of extrapersonal events in terms of their motivational salience, and to generate 'kinetic strategies' so that the attentional focus can shift from one target to another. PMID- 10466157 TI - The clinical and functional measurement of cortical (in)activity in the visual brain, with special reference to the two subdivisions (V4 and V4 alpha) of the human colour centre. AB - We argue below that, at least in studying the visual brain, the old and simple methods of detailed clinical assessment and perimetric measurement still yield important insights into the organization of the visual brain as a whole, as well as the organization of the individual areas within it. To demonstrate our point, we rely especially on the motion and colour systems, emphasizing in particular how clinical observations predicted an important feature of the organization of the colour centre in the human brain. With the use of data from functional magnetic resonance imaging analysed by statistical parametric mapping and independent component analysis, we show that the colour centre is composed of two subdivisions, V4 and V4 alpha the two together constituting the V4 complex of the human brain. These two subdivisions are intimately linked anatomically and act cooperatively. The new evidence about the architecture of the colour centre might help to explain why the syndrome, cerebral achromatopsia, produced by lesions in it is so variable. PMID- 10466158 TI - Meat or wheat for the next millennium? Proceedings of a symposium. Guildford, United Kingdom, 29 June-2 July 1998. PMID- 10466156 TI - Functional neuroimaging in psychiatry. AB - Functional neuroimaging is one of the most powerful means available for investigating the pathophysiology of psychiatric disorders. In this review, we shall focus on the different ways that it can be employed to this end, describing the major findings in the field in the context of different methodological approaches. We will also discuss practical issues that are particular to studying psychiatric disorders and the potential contribution of functional neuroimaging to future psychiatric research. PMID- 10466160 TI - Alternative futures for world cereal and meat consumption. AB - Fundamental changes in the global structure of food demand will lead to an extraordinary increase in the importance of developing countries in global food markets. Economic growth in developing countries is changing consumption patterns, with slower growth (and in many countries actual declines) in per capita food consumption of grains and rapidly growing per capita and total meat consumption, combined with induced growth in cereal feed consumption. The present paper examines the hypothesis, suggested by some researchers, that high-meat diets in developed countries limit improvement in food security in developing countries. These analysts argue that reduced meat consumption in developed countries would release cereals from livestock feed to food for poorer populations, thus improving food security in developing countries. Using the International Food Policy Research Institute (Washington, DC, USA) global food projections model, the international model for policy analysis of agricultural commodities and trade (see Rosegrant et al. 1995), we first analyse the implications for future global cereal and meat supply and demand resulting from changes in global income, population growth and other structural changes, then simulate alternative scenarios to examine the effect of large reductions in meat consumption in developed countries on food consumption and food security in developing countries. The paper shows that while the long-term prospects for food supply, demand and trade indicate a strengthening of world cereal and livestock markets, the improvement in food security in the developing world will be slow, and changes in the dietary patterns in developed countries are not an effective route to improvement in food security in developing countries. PMID- 10466159 TI - Animal v. plant foods in human diets and health: is the historical record unequivocal? AB - An ideal diet is one that promotes optimal health and longevity. Throughout history, human societies have developed varieties of dietary patterns based on available food plants and animals that successfully supported growth and reproduction. As economies changed from scarcity to abundance, principal diet related diseases have shifted from nutrient deficiencies to chronic diseases related to dietary excesses. This shift has led to increasing scientific consensus that eating more plant foods but fewer animal foods would best promote health. This consensus is based on research relating dietary factors to chronic disease risks, and to observations of exceptionally low chronic disease rates among people consuming vegetarian, Mediterranean and Asian diets. One challenge to this consensus is the idea that palaeolithic man consumed more meat than currently recommended, and that this pattern is genetically determined. If such exists, a genetic basis for ideal proportions of plant or animal foods is difficult to determine; hominoid primates are largely vegetarian, current hunter gatherer groups rely on foods that can be obtained most conveniently, and the archeological record is insufficient to determine whether plants or animals predominated. Most evidence suggests that a shift to largely plant-based diets would reduce chronic disease risks among industrialized and rapidly industrializing populations. The accomplish this shift, it will be necessary to overcome market-place barriers and to develop new policies that will encourage greater consumption of fruits, vegetables and grains as a means to promote public health. PMID- 10466161 TI - Animal- and plant-food-based diets and iron status: benefits and costs. AB - Fe seems to be the only nutrient deficiency that industrialized and low-income countries have in common. Thus, Fe is one of the most critical nutrition requirements to be met in most diets in human subjects. Fe deficiency is caused not only by too low an intake, but is also the result of low bioavailability, as well as an increased Fe requirement due to physiological variables or clinical problems which are not met by an increased dietary intake of Fe. In low-income countries poor dietary quality rather than Fe intake seems to be the key determinant of impaired Fe status. Sometimes the Fe intake even exceeds that in populations of industrialized countries. The interaction of all enhancers (e.g. ascorbic acid and meat), as well as inhibitors (such as bran, polyphenols, egg yolk, soyabean products, Ca, Ca3(PO4)2 and phytic acid (or phytate)) is what determines the bioavailability of non-haem-Fe in the meal. Dietary composition seems to be particularly important when Fe reserves are low, or in the presence of Fe deficiency. Furthermore, the development of anaemia as a result of Fe deficiency, secondary to Fe-stress situations, is dependent on the Fe balance in the host. With respect to the dietary intake of Fe, other products in the food consumed as well as previous treatment of the product (e.g. heat treatment and processing) may also influence bioavailability. Despite all efforts to counteract Fe deficiency it still represents one of the dominant problems in the micronutrient sphere. It is apparent that there is no simple solution to the problem, and the fact that Fe deficiency still occurs in affluent societies consuming a mixed diet speaks for itself a more holistic view of total dietary composition and the role of enhancers and inhibitors is needed. PMID- 10466162 TI - High-meat diets and cancer risk. AB - Up to 80% of breast, bowel and prostate cancers are attributed to dietary practices, and international comparisons show strong positive associations with meat consumption. Estimates of relative risk obtained from cohort investigations are in the same direction, although generally weak, and red and processed meats rather than white meat seem to be associated with elevated risk of colon cancer. In breast cancer, there are consistent associations with total meat intake and there is evidence of a dose response. Despite these associations with meat, existing studies suggest that vegetarians do not have reduced risk of breast, bowel or prostate cancer, but there are no quantitative estimates of amounts of meat consumed by meat eaters in these cohort studies. Possible mechanisms underlying epidemiological associations include the formation of heterocyclic amines in meat when it is cooked. These heterocyclic amines require acetylation by P450 enzymes, and individuals with the fast-acetylating genotype who eat high amounts of meat may be at increased risk of large-bowel cancer. NH3 and N-nitroso compounds (NOC) formed from residues by bacteria in the large bowel and probably also important. NH3 is a promotor of large-bowel tumours chemically induced by NOC, and some of the chromosomal mutations found in human colo-rectal cancer are consistent with effects of NOC and heterocyclic amines. However, the type, amount, and cooking method of meat or protein associated with increased risk are not certain. The effects of high levels of meat on NH3 and NOC output are not reduced by increasing the amount of fermentable carbohydrate in the diet, but interaction between meat, NSP and vegetable intakes on the risk of cancer has not been studied comprehensively. The interaction between dietary low-penetrance genetic polymorphic and somatic mutation factors has also been investigated to a limited extent. Current Department of Health (1998) recommendations are that meat consumption should not rise, and that consumers at the top end of the distribution should consider a reduction in intakes. PMID- 10466163 TI - The nutritional value of plant-based diets in relation to human amino acid and protein requirements. AB - The adequacy of plant-based diets in developed and developing countries as sources of protein and amino acids for human subjects of all ages is examined. Protein quantity is shown not to be an issue. Digestibility is identified as a problem for some cereals (millet (Panicum miliaceum) and sorghum (Sorghum sp.)) and generally is poorly understood. Direct measurements of biological value in children are reviewed and scoring is considered. Various existing requirement values for amino acids and especially lysine are reviewed, and it is concluded that stable-isotope studies do not yet provide adequate alternative values of N balance data, which for lysine are robust after recalculation and adjustment. A new maintenance requirement pattern is developed, with higher values than those of Food and Agriculture Organization/World Health Organization/United Nations University (1985) but lower values than the Massachusetts Institute of Technology pattern (Young et al. 1989). Calculations of age-related amino acid requirements are based on most recent estimates of human growth and maintenance protein requirements, a tissue amino acid pattern and the new maintenance amino acid pattern. These values appear valid when used to score plant proteins, since they indicate values similar to or less than the biological value measured directly in young children. When used to score plant-based diets in India, no marked deficiencies are identified. All regions score > 1 for adults, whilst for children scores range from > 1, (Tamil Nadhu) from 6 months of age to 0.78 (West Bengal), rising to 0.9 in the 2-5 year old, consistent with reports that high lysine maize supports similar weight and height growth to that of casein. Inadequate amino acid supply is not an issue with most cereal-based diets. PMID- 10466164 TI - Meat and colo-rectal cancer. AB - In early epidemiological studies of diet and cancer the stress was on the search for causal factors. Population (ecological) studies tended to show a strong correlation between meat intake, particularly red meat, and the risk of colo rectal cancer. They also tended to show meat to be strongly inversely correlated with cancers of the stomach and oesophagus and liver. Early case-control studies tended to support the postulated role for red meat in colo-rectal carcinogenesis, although more recent case-control studies, particularly those from Europe, have tended to show no relationship. The cohort studies in general failed to detect any relationship between meat intake and colo-rectal cancer risk. The available evidence points to the intake of protective factors such as vegetables and whole grain cereals being the main determinants of colo-rectal cancer risk, with meat intake only coincidentally related. PMID- 10466165 TI - The nutritional adequacy of plant-based diets. AB - The nutritional adequacy of plant-based diets is discussed. Energy and protein intakes are similar for plant-based diets compared with those containing meat. Fe and vitamin B12 are the nutrients most likely to be found lacking in such diets. Bioactive substances present in foods of plant origin significantly influence the bioavailability of minerals and requirements for vitamins. Well-balanced vegetarian diets are able to support normal growth and development. It is concluded that meat is an optional rather than an essential constituent of human diets. PMID- 10466166 TI - Health benefits of a vegetarian diet. AB - Compared with non-vegetarians, Western vegetarians have a lower mean BMI (by about 1 kg/m2), a lower mean plasma total cholesterol concentration (by about 0.5 mmol/l), and a lower mortality from IHD (by about 25%). They may also have a lower risk for some other diseases such as constipation, diverticular disease, gallstones and appendicitis. No differences in mortality from common cancers have been established. There is no evidence of adverse effects on mortality. Much more information is needed, particularly on other causes of death, other morbidity including osteoporosis, and long-term health in vegans. The evidence available suggests that widespread adoption of a vegetarian diet could prevent approximately 40,000 deaths from IHD in Britain each year. PMID- 10466167 TI - Childhood leukaemia: a model of pre-obesity. AB - The prevalence of obesity in children, as in adults, is increasing dramatically. The extent to which this is due to reduced energy expenditure, increased energy intake, or both, is unclear at present. This in part reflects the limitations of existing models of the pre-obese state. In childhood acute lymphoblastic leukaemia (ALL), patients typically gain weight excessively during and after 2 years of therapy, and are at high risk of becoming obese. Previous studies have failed to identify the cause of obesity in these patients. We have tested the hypothesis that excess weight gain in ALL is due to reduced total energy expenditure (TEE), measured using the doubly-labelled water method, and have identified risk factors for excess weight gain in ALL. Pre-obese children with ALL in the dynamic phase of weight gain are less physically active than their peers, with a reduced TEE of approximately 1.2 (95% CI 0.2, 2.2) MJ/d. While other factors might contribute to excess weight gain, lifestyle (i.e. reduced habitual physical activity) plays a central role in ALL. Several considerations suggest that ALL might be a useful model of the pre-obese state: lifestyle is critical to development of obesity in ALL; ALL is relatively common; approximately 70% of patients survive; patients are readily accessible during the 2 years of therapy and beyond. PMID- 10466168 TI - The influence of maternal nutrient restriction in early to mid-pregnancy on placental and fetal development in sheep. AB - Placental weight is a primary factor determining size at birth in many species. In sheep, placental weight peaks at approximately mid-gestation, with structural remodelling occurring over the second half of pregnancy to meet the increasing nutritional demands of the growing fetus. Numerous factors influence placental growth and development in sheep, and many workers (see Kelly, 1992) have investigated the role of maternal nutrition as a regulator of placental and fetal size. We have studied the effects of feeding ewes approximately 50% of their recommended energy requirements during early to mid-pregnancy on fetal and placental indices measured at mid-gestation (i.e. 80 d) and close to term (i.e. 145 d). Maternal nutrient restriction is associated with a reduction in placental weight at 80 d, but an increase in placental weight at 145 d of gestation, compared with ewes fed adequately in early pregnancy. No significant effect on fetal weight was observed at either 80 or 145 d gestation, although differences in body dimensions and the insulin-like growth factor-1 axis were found in lambs from nutrient-restricted ewes delivered close to term. Maternal nutrition during pregnancy plays a pivotal role in the regulation of fetal and placental development in sheep, and therefore has the potential to influence both short- and longer-term health outcomes. PMID- 10466169 TI - The effect of vitamin A on epithelial integrity. AB - Vitamin A is the generic term for a variety of fat-soluble substances including retinol, retinyl palmitate and the provitamin A carotenoids such as all-trans beta-carotene. Vitamin A is commonly known as the anti-infective vitamin and has an essential role in vision and cellular differentiation, the latter providing a unique core mechanism helping to explain the influence of vitamin A on epithelial barriers. Alterations in the epithelial lining of vital organs occur early in deficiency, suggesting a potentially important role for the barrier function. Vitamin A deficiency (VAD) is most commonly recognized in the eye. The conjunctival-impression cytology test detects the presence of larger irregular keratinized cells and the absence of mucous-secreting goblet cells, indicative of VAD. The method is simple, quick and sensitive in populations where VAD is present. In the respiratory tract, observational studies all show an association with VAD, although vitamin A supplementation studies appear to have little effect on respiratory disease. Organ-specific targeting may improve success rates. The dual-sugar intestinal-permeability test allows the effect of vitamin A supplementation to be monitored on the gastrointestinal tract. Two vitamin A supplementation studies were carried out recently in Orissa State, India. Healthy infants of weaning age were administered orally eight weekly doses of 5.0 mg retinol equivalents and hospitalized infants received one large oral dose 60 mg retinol equivalents in the form of retinyl palmitate. Improvements in gut integrity and haematological status were observed in both studies. In summary, the response of the eye to vitamin A supplementation is well established; the present review highlights some of the more recent observations examining the effects of vitamin A. PMID- 10466170 TI - A possible role for vitamin C in age-related cataract. AB - While many experimental studies have shown a protective effect of vitamin C in age-related cataract, other studies have revealed contrasting roles for this nutrient. Oxidative damage in the lens can be prevented by vitamin C. However, a pro-oxidant effect of vitamin C through H2O2 generation has been suggested. Vitamin C has also been shown to play a role in protein glycation, which is observed in cataract formation. A protective effect of dietary energy restriction appears to be inversely related to plasma vitamin C levels in rodents. Moreover, conclusions from human epidemiological and intervention studies are not uniform. The available evidence suggests that maintenance of sufficient plasma vitamin C is needed to prevent oxidative damage in the lens. More research will be needed in order to confirm the relative importance of of the different roles of vitamin C in the eye lens. PMID- 10466171 TI - Nutrition and lung health. AB - There is an increasing interest in the relationship between nutrition and lung health. Epidemiological studies suggest that dietary habits may have an influence on lung function and the tendency to common lung diseases such as asthma, chronic obstructive pulmonary disease (COPD) and lung cancer. In particular, a diet rich in fresh fruit and fish has been associated with a salutary effect on lung health. End-stage COPD is associated with a state of nutritional depletion which is refractory to conventional nutritional supplementation. In contrast, malnutrition associated with cystic fibrosis is amenable to nutritional therapy, which has been shown to improve prognosis in this disease. PMID- 10466172 TI - Dietary influences on chronic obstructive lung disease and asthma: a review of the epidemiological evidence. AB - The epidemiological evidence for a relationship between diet and indicators of asthma and chronic obstructive pulmonary disease (COPD) is evaluated. The review focuses on the intake of Na, n-3 fatty acids, and antioxidant vitamins as well as fruit and vegetables. Experimental studies suggest that a high-Na diet has a small adverse effect on airway reactivity in asthma patients. However, observational studies provide no clear evidence that high Na intake has adverse effects on airway reactivity or asthma symptoms in open populations. n-3 Polyunsaturated fatty acids, which are present in fish oils, are metabolized into less broncho-constricting and inflammatory mediators than n-6 polyunsaturated fatty acids. Studies in the general adult population suggest that a high fish intake has a beneficial effect on lung function, but the relationship with respiratory symptoms and clinically-manifest asthma or COPD is less evident. Also, experimental studies in asthma patients have not demonstrated an improvement in asthma severity after supplementations with fish oil. Several studies showed a beneficial association between fruit and vegetable intake and lung function, but the relationship with respiratory symptoms and the clinically manifest disease was less convincing. A similar pattern was found for vitamin C in relation to indicators of asthma and COPD, but there are still conflicting results with respect to vitamin E and beta-carotene. In conclusion, the epidemiological evidence for a beneficial effect on indicators of asthma and COPD of eating fish, fruit and vegetables is increasing. However, the effectiveness of dietary supplementation in open-population samples is often not demonstrated. Several unresolved questions are raised, which should be addressed in future studies on the relationship between diet and respiratory disease. PMID- 10466173 TI - The pulmonary cachexia syndrome: aspects of energy balance. AB - The present paper reviews current knowledge of the pulmonary cachexia syndrome with reference to chronic obstructive pulmonary disease (COPD). Aspects of incidence, aetiology and management are discussed. Malnutrition occurs in approximately one-quarter to one-third of patients with moderate to severe COPD. Both fat mass and fat-free mass become depleted. Loss of fat-free mass is the more important and appears to be due to a depression of protein synthesis. Weight loss is an independent prognostic indicator of mortality, and is associated with increased morbidity and decreased health-related quality of life. The aetiology of malnutrition in COPD is not well understood. Reduced food intake does not seem to be the primary cause. Resting energy expenditure (REE) is elevated in a proportion of patients and probably contributes to negative energy balance. Measurement of actual REE is helpful when considering the adequacy of nutritional supplementation. The underlying reason for a hypermetabolic state is not known. Although weight-losing COPD patients are not catabolic, nutritional supplementation alone does not appear to reverse the loss of fat-free mass. Strategies involving nutritional supplementation in combination with a second intervention are being explored, and there are some encouraging results using anabolic hormones. PMID- 10466174 TI - Vitamins and lung cancer. AB - Lung cancer is the most common cancer in the world, and smoking is the major risk factor, accounting for about 90% of the cases. Diet has also been implicated in the development of lung cancer, although the specific nutrients remain to be elucidated. Vitamins with antioxidant activity have received much attention. beta Carotene, the most efficient provitamin A, was found to be inversely related to the risk of lung cancer in many prospective epidemiological studies, especially in studies measuring serum concentrations of beta-carotene. The findings from controlled trials, however, contradict the hypothesis that beta-carotene could prevent lung cancer, but rather suggest increased risk of lung cancer with supplementary beta-carotene. Data from both prospective studies and a controlled trial suggest no role for vitamin E in lung carcinogenesis. Some prospective epidemiological studies suggest an inverse relationship between dietary vitamin C and the risk of lung cancer, but due to the high correlation between dietary vitamin C and vegetable and fruit intake the independent role of dietary vitamin C is difficult to estimate. Studies using prediagnostic plasma concentrations of ascorbic acid do not support the involvement of vitamin C in lung carcinogenesis, and no controlled trials of vitamin C on lung cancer have been published. Thus, supplementation with antioxidant vitamins cannot be recommended for the prevention of lung cancer. Non-smoking is the most important target in the prevention of lung cancer. High intakes of vegetables and fruits may provide additional protection and are unlikely to be harmful. PMID- 10466175 TI - Diet, health and globalization: five key questions. AB - The present paper explores possible implications of the globalization of the food system for diet and health. The paper poses five key questions to clarify the relationship between food and globalization. The first question is what is globalization. The paper suggests that it is helpful to distinguish between economic, political, ideological and cultural processes. Globalization is also marked by internal oppositional dynamics: there are re-localization and regional tendencies which counter the global. The second question is whether there is anything new about globalization. Food has been a much traded commodity for millennia. The paper concludes that what is new about the current phases of globalization is the pace and scale of the change, and the fact that power is being concentrated into so few hands. New marketing techniques and supply-chain management consolidate these features. The third question is who is in control of the globalization era and who benefits and loses from the processes of globalization. It is argued that modern food economies are hypermarket rather than market economies, with power accruing to the distributor more than has been recognized. The fourth question concerns governance of the food system. Historically, systems of local and national government have regulated the food supply where appropriate. Now, new international systems are emerging, partly using existing bodies and partly creating new ones. The final question is of the future. Globalization is a value-laden area of study, yet its implications for dietary change and for health are considerable. The paper argues that dimensions of change can be discerned, although it would be rash to bet on which end of each dimension will emerge as dominant in the 21st century. PMID- 10466176 TI - China: the soyabean-pork dilemma. AB - In 1996 the population of China reached 1.23 billion, 22% of the world population, and is expected to increase to 1.5 billion by 2020. As China has only 7% of the world's arable land such population increases are likely to have an important impact on food supply in China and the world. Projections of the potential impact are discussed. The restructuring of Chinese agriculture at the end of the 1970s has led to dramatic increases in agricultural production and food consumption, in particular of animal products, fruit and vegetables. Along with these rapid changes there is evidence of a nutrition transition in which diseases associated with affluence are becoming more prevalent than deficiency diseases. This transition has led to concern about the evolving dietary pattern. The replacement of legumes, including soyabean, by meat and other animal products as rich sources of protein and other nutrients has been controversially argued on grounds of nutritional health, ecological impact, economic effects and world food supply. These arguments are reviewed and the pressures internal and external to China concerning the production and consumption of animal v. legume products are presented. It is concluded that nutritional policies to promote the consumption of soyabean are unlikely to be effective in the context of an increasingly free and global market. PMID- 10466177 TI - Food aid in emergencies: a case for wheat? AB - As disasters and conflict increase, a higher proportion of total food aid is given as humanitarian aid. Most food aid is in the form of cereals, primarily wheat. The main donors are the USA and the EU, but there is an increase in the numbers of donors, including non-governmental organizations, buying food rather than using surpluses. Alongside the greater diversity and complexity of food aid, there is more controversy about policy and practice. If disasters are development failures, emergency food aid must be a step in the continuum from relief to rehabilitation. Comparisons of the seventeen countries that were major recipients of food aid (> 10,000 t) in 1997, show diversity in social development, dietary pattern, number of refugees, relative food inadequacy and wasting (i.e. % standard weight-for-height > 2 SD). In the absence of information of consistent quality, what influences the scale of emergency aid is unclear and susceptible to politicization, so that need and supply may not be matched. Local considerations seem to be as important as external food aid for the nutrition of the recipients. Challenges for the future include assuring the nutritional quality of rations to solve deficiency problems. The implications for the professional public health nutritionist working on emergency food provision include continuing professional development to enhance the technical expertise necessary to design appropriate feeds or rations. These public health nutritionists, more than others, require a grounding in social science theories that underpin management, ethics of professionalism and the politics of food aid. PMID- 10466178 TI - Manipulating meat quality and composition. AB - Meat quality describes the attractiveness of meat to consumers. The present paper focuses on two major aspects of meat quality, tenderness and flavour. Both aspects of quality can be influenced by nutrition, principally through its effects on the amount and type of fat in meat. In several countries, high levels of intramuscular fat (marbling fat), i.e. above 30 g/kg muscle weight in longissimus, are deemed necessary for optimum tenderness, although poor relationships between fat content and tenderness have generally been found in European studies, where fat levels are often very low, e.g. below 10 g/kg in UK pigs. Muscle lipid may be a marker for red oxidative (type 1) muscle fibres which are found at higher concentrations in tender muscles and carcasses. Nutritional treatment can be used to manipulate the fatty acid content of muscle to improve nutritional balance, i.e. increase the polyunsaturated (PUFA): saturated fatty acid value and reduce the n-6:n-3 PUFA value. Increasing PUFA levels may also change flavour because of their greater susceptibility to oxidative breakdown and the generation of abnormal volatile compounds during cooking. This situation particularly applies to the n-3 PUFA which are the most unsaturated meat lipids. In pigs, a concentration of 3 mg alpha-linolenic acid (18:3)/100 mg in muscle and fat tissue fatty acids can easily be achieved by including whole linseed in the diet. This level has led to abnormal odours and flavours in some studies, but not in others. In cattle and sheep, feeding whole linseed raised 18:3 concentrations in muscle fatty acids from about 0.7 mg/100 mg to > 1 mg/100 mg. As with pigs, this diet also increased levels of long-chain n-3 PUFA formed from 18:3, including eicosapentaenoic acid (20:5). Although this increase led to greater oxidative breakdown of lipids during storage and the generation of large quantities of lipid-derived volatile compounds during cooking, there were no deleterious effects on odour or flavour. When 18:3 levels are raised in lamb and beef because of grass feeding, the intensity of the flavours increases in comparison with grain-fed animals which consume and deposit relatively more linoleic acid (18:2). In ruminants, very high levels of 18:2 produced by feeding protected oil supplements cause the cooked beef to be described as oily, bland or pork-like. PMID- 10466179 TI - Meat production in developing countries. AB - Developing countries have very diverse food consumption patterns and agricultural production systems. The proportion of meat in national diets varies from negligible in some countries in central Africa to 30-40% in some countries in Latin America and Mongolia. However, the demand for meat in developing countries is increasing rapidly (53%/year from 1982 to 1993), as the result of population growth and the trend for people to move to the cities. Growth rates in consumption are greatest in Asia, with China dominating the statistics, in view of the size of its population. Theoretically, livestock production can be increased to meet this demand, but the multiple roles of livestock in developing countries must be recognized if this is to be achieved in a sustainable manner. Resource-poor farmers who keep livestock may value more highly their contribution to livelihoods and to crop production, through provision of draught power and improvements in soil fertility through the recycling of manure, than the production of more meat. Recognition of the goals of the farmer and the wishes of the consumer regarding meat quality need to be reflected in the way in which opportunities for increasing meat production are identified and communicated to farmers. The impact of the global economy on cereal prices, for example, will also influence which interventions will be economically viable. Interpretation of information in an integral manner, using geographical information systems, mathematical models and/or simple spreadsheet models will be an important ingredient in turning scientific knowledge into increased meat production in developing countries. PMID- 10466180 TI - Development of farmed fish: a nutritionally necessary alternative to meat. AB - The projected stagnation in the catch from global fisheries and the continuing expansion of aquaculture is considered against the background that fishmeal and fish oil are major feed stocks for farmed salmon and trout, and also for marine fish. The dietary requirement of these farmed fish for high-quality protein, rich in essential amino acids, can be met by sources other than fishmeal. However, the highly-polyunsaturated fatty acids eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3) present in high concentrations in fish oil are essential dietary constituents for marine fish and highly-desirable dietary constituents for salmonids. Currently, there is no feasible alternative source to fish oil for these nutrients in fish feeds. Vegetable oils rich in linoleic acid (18:2n-6) can partially substitute for 20:5n-3 and 22:6n-3 in salmonid and marine fish feeds. However, this is nutritionally undesirable for human nutrition because the health-promoting effects of fish-derived 20:5n-3 and 22:6n-3 reflect a very high intake of 18:2n-6 relative to linolenic acid (18:3n-3) in Western diets. If partial replacement of fish oils in fish feeds with vegetable oils becomes necessary in future, it is argued that 18:3n-3-rich oils, such as linseed oil, are the oils of choice because they are much more acceptable from a human nutritional perspective, especially given the innate ability of freshwater fish, including salmonids, to convert dietary 18:3n-3 to 20:5n-3 and 22:6n-3. In the meantime, a more judicious use of increasingly-expensive fish oil in aquaculture is recommended. High priorities in the future development of aquaculture are considered to be genetic improvement of farmed fish stocks with enhanced abilities to convert C18 to C20 and C22 n-3 polyunsaturated fatty acids, enhanced development of primary production of 20:5n-3 and 22:6n-3 by single-cell marine organisms, and continuing development of new species. PMID- 10466181 TI - New understanding in obesity research. AB - A public-health approach considers the relevance of nutritional research in the prevention and management of obesity. Well-defined and internationally-agreed definitions based on BMI allow an assessment of the worldwide prevalence of overweight and obesity. There are about 250 million obese adults in the world, and many more overweight. Obesity is emerging in the Third World, first in urban middle-aged women. With economic developments, obesity then occurs in men and younger women. In the West childhood obesity is rapidly emerging, with concern that early-onset obesity is especially hazardous. In Asians the risks of excess visceral fat occur at lower body weights than in Caucasians. The propensity to visceral obesity in Asians may relate to malnourished mothers and low birth weight. The International Obesity Task Force is considering many issues, including the health economics of obesity. It has developed a strategy to define childhood obesity, which in children over 6 years is likely to predict long-term weight and health problems. While the search for genetic markers of obesity continues, with particular interest in the leptin gene, it is clear that societal change, with the decline in physical activity and the passive overconsumption of high-fat diets are major contributors to the global increase in obesity. The public-health aspects of obesity research are therefore challenging. PMID- 10466182 TI - Optimal nutrition. Proceedings of a symposium. Cork, United Kingdom, 22-24 July 1998. PMID- 10466183 TI - Unsaturated fatty acids. AB - There is good scientific evidence that dietary fatty acid composition is involved in the aetiology of many diseases. Increasing the supply of n-3 polyunsaturated fatty acids (PUFA) may reduce the risk of CHD. Several scientific organizations (for example, see Department of Health, 1991, 1994; British Nutrition Foundation, 1992; Scientific Committee for Food, 1993; Food and Agriculture Organization/World Health Organization, 1998) have made recommendations for n-3 PUFA; however, there is a high degree of variation both in terms of the type and amount of n-3 PUFA (up to 7-fold). This variation reflects the different scientific axioms which underlie the different recommendations. Optimal nutrition may be defined in terms of the level of a nutrient required to avoid deficiency, or the amount required to have an effect on biomarkers and functional indicators of nutrient intake, or the level of a nutrient which prevents disease. Functional biomarkers of n-3 PUFA include plasma, platelet and erythrocyte phospholipid-n-3 PUFA levels. Plasma triacylglycerol concentrations represent a functional indicator of n-3 PUFA because n-3 PUFA exert a consistent hypotriacylglycerolaemic effect which is dose-dependent and persistent. In terms of disease status, epidemiological studies have demonstrated that the incidence of CHD is inversely associated with consumption of n-3 PUFA. Despite the health benefits of n-3 PUFA, the mean daily intake falls far short of most of the recommendations. Increasing fish intake is the most obvious way to increase n-3 PUFA intake. However, a large percentage (up to 65) of the population do not eat fish. Thus, there is a need for alternative sources of n-3 PUFA, such as functional foods, whose unique fatty acid composition could fortify staple foods thereby promoting optimal levels of n-3 PUFA intake. PMID- 10466184 TI - Optimal intakes of protein in the human diet. AB - For protein, progress is slow in defining quantifiable indicators of adequacy other than balance and growth. As far as current requirements are concerned, only in the case of infants and children is there any case for revision, and this change is to lower values. Such intakes would appear to be safe when consumed as milk formula. In pregnancy, notwithstanding the concern that deficiency may influence programming of disease in later life, there is little evidence of any increased need, and some evidence that increased intakes would pose a risk. For the elderly there is no evidence of an increased requirement or of benefit from increased intakes, except possibly for bone health. For adults, while we now know much more about metabolic adaptation to varying intakes, there would appear to be no case for a change in current recommendations. As far as risks and benefits of high intakes are concerned, there is now only a weak case for risk for renal function. For bone health the established views of risk of high protein intakes are not supported by newly-emerging data, with benefit indicated in the elderly. There is also circumstantial evidence for benefit on blood pressure and stroke mortality. With athletes there is little evidence of benefit of increased intakes in terms of performance, with older literature suggesting an adverse influence. Thus, given that a safe upper limit is currently defined as twice the reference nutrient intake, and that for individuals with high energy requirements this value (1.5 g/kg per d) is easily exceeded, there is a case for revising the definition of a safe upper limit. PMID- 10466185 TI - Optimal nutrition: fibre and phytochemicals. AB - There is currently intense research interest in secondary plant metabolites because of their potential preventative effects on the chronic diseases of Western societies, especially cardiovascular disease and cancer. To date most of the research has focused on the identification of plant-derived substances and their potential protective effects against specific chronic diseases. The important issue of determining the optimal intake of those substances, such that the beneficial effects are maximized without manifestation of adverse effects, has yet to be addressed in most cases. Furthermore, there are no specific functional markers that can be used to assess optimal intake, although it may be possible to use biomarkers such as serum cholesterol if the rest of the diet is strictly controlled. The present review discusses a wide range of substances associated with plants, including dietary fibre, resistant starch, oligosaccharides, phyto-oestrogens, phytosterols, flavonoids, terpenes and isothiocyanates, and attempts where possible to indicate optimal intakes and to suggest functional markers. PMID- 10466186 TI - Optimal macronutrient balance. AB - There is at present a justifiable debate as to the optimum level of total dietary fat which will reduce the risk of obesity without an elevation of plasma triacylglycerol or a depression of plasma HDL-cholesterol. Total plasma cholesterol and LDL-cholesterol levels are lowered and risk of fatal myocardial infarction is lowered when either saturated or trans-unsaturated fatty acids are replaced isoenergetically by either monounsaturated or polyunsaturated fatty acids. The triacylglycerol-raising and HDL-lowering effects of low-fat high carbohydrate diets can be overcome with low intakes of n-3 polyunsaturated fatty acids and moderate exercise. Whilst a reduction in dietary fat is being attained in many countries, the reduction is uniform across all fatty acids, leaving dietary fat composition unchanged. The ability of low-fat diets to reduce cholesterol and cause a fall in body weight is not influenced by the carbohydrate ratio starch: sugars in the diet. However, weight-gain susceptibility to high intakes of dietary fat and the plasma cholesterol responsiveness to diet are considerably influenced by common genetic polymorphisms. PMID- 10466187 TI - Optimum nutrition: thiamin, biotin and pantothenate. AB - The metabolism of glucose is deranged in thiamin deficiency, but once any deficiency has been corrected there is no further effect of increased thiamin intake on the ability to metabolize glucose through either pyruvate dehydrogenase (EC 1.2.4.1) and the citric acid cycle, or the pentose phosphate pathway, in which transketolase (EC 2.2.1.1) is the thiamin-dependent step. It has been suggested that the Wernicke-Korsakoff syndrome is associated with a genetic variant of transketolase which requires a higher than normal concentration of thiamin diphosphate for activity. This finding would suggest that there may be a group of the population who have a higher than average requirement for thiamin, but the evidence is not convincing. There are no estimates of biotin requirements, but either coenzyme saturation of erythrocyte pyruvate carboxylase, or the excretion of 3-hydroxy-isovalerate (perhaps after a test dose of leucine) could be used to assess requirements in depletion-repletion studies. Biotin deficiency leads to impaired glucose tolerance, but it is unlikely that glucose tolerance could be used to assess optimum biotin status, since other more common factors affect glucose tolerance to a greater extent. Plasma triacylglycerol and nonesterified fatty acids are moderately elevated in pantothenic acid deficiency. However, this is unlikely to be useful in assessing pantothenate status, since again, other more common factors affect plasma lipids. To date there are no biochemical indices of adequate pantothenate nutrition, and no estimates of requirements. PMID- 10466188 TI - Current knowledge concerning optimum nutritional status of riboflavin, niacin and pyridoxine. AB - The term 'optimum nutrition' has evolved from a perceived need to base recommendations for nutrient intakes firmly in the context of function. It follows that 'optimum nutritional status' for individual nutrients should be defined in terms of biochemical or physiological markers having some functional value but also showing an appropriate relationship to nutrient intake. The present short review considers the current position regarding such markers for riboflavin, pyridoxine and niacin. It is concluded that whilst there are several biochemical measures which respond to changes in intake of each of these vitamins, no single measure is wholly satisfactory as a marker of optimum status. PMID- 10466189 TI - Folate and vitamin B12. AB - The folates are made up of a pterdine ring attached to a p-aminobenzoate and a polyglutamyl chain. The active form is tetrahydrofolate which can have C1 units enzymically attached. These C1 units (as a formly group) are passed on to enzymes in the purine pathway that insert the C-2 and C-8 into the purine ring. A methylene group (-CH2-) attached to tetrahydrofolate is used to convert the uracil-type pyrimidine base found in RNA into the thymine base found in DNA. A further folate cofactor, i.e. 5-methyltetrahydrofolate, is involved in the remethylation of the homocysteine produced in the methylation cycle back to methionine. After activation to S-adenosylmethionine this acts as a methyl donor for the dozens of different methyltransferases present in all cells. Folate deficiency results in reduction of purine and pyrimidine biosynthesis and consequently DNA biosynthesis and cell division. This process is most easily seen in a reduction of erythrocytes causing anaemia. Reduction in the methylation cycle has multiple effects less easy to identify. One such effect is certainly on the nerve cells, because interruption of the methylation cycle causing neuropathy can also happen in vitamin B12 deficiency due to reduced activity of the vitamin B12-dependent enzyme methionine synthase (EC 2.1.1.13). In vitamin B12 deficiency, blocking of the methylation cycle causes the folate cofactors in the cell to become trapped as 5-methyltetrahydrofolate. This process in turn produces a pseudo folate deficiency in such cells, preventing cell division and giving rise to an anaemia identical to that seen in folate deficiency. PMID- 10466190 TI - Optimal nutrition: vitamin A and the carotenoids. AB - There are two major dietary sources of vitamin A: easily absorbed retinyl palmitate in foods of animal origin, and poorly bioavailable carotenoids from plant foods. Plasma retinol is tightly controlled, probably by regulation of retinol-binding protein (RBP) formation in the liver, and only hormonal factors (e.g. oral contraceptives) and infection will alter the homeostasis. Delivery of retinol to the tissues is facilitated by the RBP-retinol complex; however, there is evidence that this mechanism can be bypassed when very high doses of vitamin A are given. Some retinyl ester may be released to tissues from chylomicrons when the latter bind to tissue lipoprotein receptors during their passage from the gut to the liver following a meal. High-dose vitamin A therapy is a means of rapidly improving vitamin A status in persons with sub-optimal vitamin A nutrition but there are dangers of toxic symptoms (e.g. teratogenicity) from excess vitamin A usage. Evidence is presented to suggest that the plasma retinol: RBP may be a guide to optimal vitamin A status, since values less than one frequently occur in less-developed countries and during infection. In contrast to plasma retinol, plasma carotenoids reflect the dietary intake of plant foods. However, absorption is limited by poor bioavailability and a saturable uptake mechanism in competition with other phytochemicals. Recent work on bioavailability suggests that the calculation of plant food vitamin A activity should be re-examined. Illness has little influence on plasma levels except by suppressing appetite. Carotenoids are generally regarded as non-toxic yet intervention studies with beta-carotene in smokers have been associated with increased lung cancer and heart disease. Some carotenoids are important as vitamin A precursors, but the physiological importance of their antioxidant properties is not known and consequently the amount needed for optimal nutrition is uncertain. PMID- 10466191 TI - Optimal nutrition: vitamin E. AB - Interest in the role of vitamin E in disease prevention has encouraged the search for reliable indices of vitamin E status. Most studies in human subjects make use of static markers, usually alpha-tocopherol concentrations in plasma or serum. Plasma or serum alpha-tocopherol concentrations of < 11.6, 11.6-16.2, and > 16.2 mumol/l are normally regarded as indicating deficient, low and acceptable vitamin E status respectively, although more recently it has been suggested that the optimal plasma alpha-tocopherol concentration for protection against cardiovascular disease and cancer is > 30 mumol/l at common plasma lipid concentrations in combination with plasma vitamin C concentrations of > 50 mumol/l and > 0.4 mumol beta-carotene/l. Assessment of vitamin E status has also been based on alpha-tocopherol concentrations in erythrocytes, lymphocytes, platelets, lipoproteins, adipose tissue, buccal mucosal cells and LDL, and on alpha-tocopherol: gamma-tocopherol in serum or plasma. Erythrocyte susceptibility to haemolysis or lipid oxidation, breath hydrocarbon exhalation, oxidative resistance of LDL, and alpha-tocopheryl quinone concentrations in cerebrospinal fluid have been used as functional markers of vitamin E status. However, many of these tests tend to be non-specific and poorly standardized. The recognition that vitamin E has important roles in platelet, vascular and immune function in addition to its antioxidant properties may lead to the identification of more specific biomarkers of vitamin E status. PMID- 10466192 TI - Vitamin C: prospective functional markers for defining optimal nutritional status. AB - Most species of plants and animals synthesize ascorbic acid, but human subjects cannot, making vitamin C an essential component of our diet. Relationships between vitamin C intake and status, and between status and health are not yet clear. There is evidence, however, that higher intake of vitamin C is associated with lower risk of disease, supporting the concept that optimal intake is needed for optimal vitamin C status, and that both factors are required for optimal health. Vitamin C has low toxicity in healthy subjects, but a clear definition of optimal status and the dietary intake required to meet and maintain this status is needed before a change in the current recommended intake can be considered. Available evidence suggests that intake of 200 mg vitamin C/d saturates tissues and maintains fasting plasma levels above the proposed threshold (50 mumol/l) for minimum risk of CHD. However, the issue of whether or not these levels produce 'optimal vitamin C status' awaits the clear and accepted definition of the term. This definition in turn awaits the development of reliable functional markers capable of assessing the effects of varying levels of vitamin C nutriture. In the present paper the relationship between intake and body stores of vitamin C and the role of vitamin C in human health are reviewed briefly. The requirements of a reliable functional marker of human vitamin C status are defined, three classes of functional markers (molecular, biochemical and physiological) are described, and possible candidate markers are examined. PMID- 10466193 TI - Optimal nutrition: calcium, magnesium and phosphorus. AB - In the past, a major challenge for nutrition research was in defining indicators of nutritional adequacy. More recently, the research base related to the role of nutrition in chronic disease has expanded sufficiently to permit moving beyond deficiency indicators to other indicators with broader functional significance. Thus, nutrition research is faced with the new challenge of defining 'optimal nutrition'. One definition of optimal nutrition with respect to any particular nutrient could be when a functional marker reaches an 'optimal value' or plateau beyond which it is not longer affected by intake or stores of the nutrient. A functional marker of nutrient status could be defined as a physiological or biochemical factor which (1) is related to function or effect of the nutrient in target tissue(s) and (2) is affected by dietary intake or stores of the nutrient (which may include markers of disease risk). Examples of such indicators or markers are those related to risk of chronic diseases such as osteoporosis, CHD, or hypertension. The present review focuses on the concept of optimal nutrition with respect to three nutrients, Ca, Mg and P. However, for P and Mg there are as yet no functional indicators which respond to dietary intake, and in such cases nutrient requirements are established using more traditional approaches, e.g. balance data. For Ca, there has been interest in using maximal Ca retention, which is based on balance data, bone mass measurements and biomarkers of bone turnover as useful functional indicators of the adequacy of Ca intake. PMID- 10466194 TI - Defining optimal body iron. AB - The major liabilities of Fe lack include defects in psychomotor development in infants, impaired educational performance in schoolchildren, increased perinatal morbidity, and impaired work capacity. Few if any of the relevant investigations have demonstrated these abnormalities in the absence of anaemia. Consequently, adequate Fe nutrition can be defined as a normal haemoglobin concentration. On the other hand, optimal Fe nutrition should be regarded as sufficient body Fe to avoid any limitation in tissue Fe supply, termed Fe-deficient erythropoiesis. A variety of laboratory measurements have been used to identify this milder form of Fe deficiency, including serum ferritin, transferrin saturation, erythrocyte protoporphyrin, mean corpuscular volume, and more recently the concentration of the soluble fragment of transferrin receptor in serum. Recent studies indicate that the serum transferrin receptor is the preferred measurement, because enhanced synthesis of the transferrin receptor represent the initial cellular response to a declining Fe supply. Moreover, unlike other methods, it is not affected by chronic inflammation or infection which are often confused with Fe deficiency. In an otherwise normal healthy population the transferrin receptor: ferritin value provides a useful quantitative index of body Fe over a wide spectrum of Fe status, ranging from Fe repletion to Fe-deficiency anaemia. It is concluded that optimal Fe nutrition is best defined as a normal haemoglobin, serum ferritin and transferrin receptor concentration. PMID- 10466195 TI - Considerations for determining 'optimal nutrition' for copper, zinc, manganese and molybdenum. AB - Defining optimal dietary intakes of Cu and Zn throughout the life cycle continues to present a considerable challenge for nutrition scientists. Although the daily intake of these micronutrients is below that currently recommended for many groups, traditional biochemical indicators of nutritional status for these trace metals largely remain within the normal range. Thus, it is unclear whether the recommended daily intakes may be unnecessarily high, or if the commonly-used markers simply lack the necessary sensitivity and specificity that are required for accurately assessing Cu and Zn status. The increasing number of reports that daily supplements with these trace metals enhance the activities of selective metalloenzymes and specific cellular and organ processes further points out the need to differentiate between meeting the requirement and providing optimal nutriture. Results from recent studies suggesting that alternative molecular and functional markers possess sufficient sensitivity to better assess Cu and Zn status are discussed. Likewise, recent studies evaluating the impact of very low and excessive levels of dietary Mn and Mo on selective biochemical and metabolic indicators are reviewed. PMID- 10466196 TI - Functional indicators of iodine and selenium status. AB - The micronutrient I, which has only one known biological function as a component of thyroid hormones, contrasts with Se, which is essential for many biochemical pathways through a range of Se-containing proteins. Thus, for I it is less complicated to propose and validate biochemical or functional markers which represent the adequacy or otherwise of dietary intake than it is to perform the same task for Se. Plasma thyroid-stimulating hormone provides a good indicator of functional I status, since levels of the hormone increase to compensate for any inadequacy in available dietary I. This increase occurs even when problems are caused by goitrogens restricting utilization of dietary I that otherwise would be adequate. In contrast, there are many potential indicators of Se status, especially since the many Se-containing proteins relate to different biochemical functions in the body. However, determination of the different GSH peroxidase (EC 1.11.1.9) activities in blood or tissue may give a useful indication of 'functional Se status', since these selenoproteins are often the most sensitive indicators of dietary Se intake. This approach has been successful in many experiments where animals of a similar genetic background have consumed diets which differ only in their Se content. Unfortunately, use of indicators of Se status in human populations may be complicated by differences (possibly genetic) between individuals that increase variability of selenoenzyme activities. The most appropriate indicator of functional Se status may also vary with other dietary conditions such as vitamin E or I deficiencies which may interact with Se deficiency. However, taking particular circumstances into account, determination of blood or tissue selenoprotein activity can provide a useful marker of adequacy of dietary Se supply for maintaining optimal health. PMID- 10466197 TI - Transformation and mineralization of 2,4,6-trinitrotoluene (TNT) by manganese peroxidase from the white-rot basidiomycete Phlebia radiata. AB - The degradation of the nitroaromatic pollutant 2,4,6-trinitrotoluene (TNT) by the manganese-dependent peroxidase (MnP) of the white-rot fungus Phlebia radiata and the main reduction products formed were investigated. In the presence of small amounts of reduced glutathione (10 mM), a concentrated cell-free preparation of MnP from P. radiata exhibiting an activity of 36 nkat/ml (36 nmol Mn(II) oxidized per sec and per ml) transformed 10 mg/l of TNT within three days. The same preparation was capable of completely transforming the reduced derivatives of TNT. When present at 10 mg/l, the aminodinitrotoluenes were transformed in less than two days and the diaminonitrotoluenes in less than three hours. Experiments with 14C-U-ring labeled TNT and 2-amino-4,6-dinitrotoluene showed that these compounds were mineralized by 22% and 76%, respectively, within 5 days. Higher concentrations of reduced glutathione (50 mM) led to a severe inhibition of the degradation process. It is concluded that Phlebia radiata is a good candidate for the biodegradation of TNT as well as its reduction metabolites. PMID- 10466198 TI - Biodegradation of 2-methyl, 2-ethyl, and 2-hydroxypyridine by an Arthrobacter sp. isolated from subsurface sediment. AB - A bacterium capable of degrading 2-methylpyridine was isolated by enrichment techniques from subsurface sediments collected from an aquifer located at an industrial site that had been contaminated with pyridine and pyridine derivatives. The isolate, identified as an Arthrobacter sp., was capable of utilizing 2-methylpyridine, 2-ethylpyridine, and 2-hydroxypyridine as primary C, N, and energy sources. The isolate was also able to utilize 2-, 3-, and 4 hydroxybenzoate, gentisic acid, protocatechuic acid and catechol, suggesting that it possesses a number of enzymatic pathways for the degradation of aromatic compounds. Degradation of 2-methylpyridine, 2-ethylpyridine, and 2 hydroxypyridine was accompanied by growth of the isolate and release of ammonium into the medium. Degradation of 2-methylpyridine was accompanied by overproduction of riboflavin. A soluble blue pigment was produced by the isolate during the degradation of 2-hydroxypyridine, and may be related to the diazadiphenoquinones reportedly produced by other Arthrobacter spp. when grown on 2-hydroxypyridine. When provided with 2-methylpyridine, 2-ethylpyridine, and 2 hydroxypyridine simultaneously, 2-hydroxypyridine was rapidly and preferentially degraded; however there was no apparent biodegradation of either 2-methylpyridine or 2-ethylpyridine until after a seven day lag. The data suggest that there are differences between the pathway for 2-hydroxypyridine degradation and the pathways(s) for 2-methylpyridine and 2-ethylpyridine. PMID- 10466199 TI - Batch culture biodegradation of methylhydrazine contaminated NASA wastewater. AB - The batch culture degradation of NASA wastewater containing mixtures of citric acid, methylhydrazine, and their reaction product was studied. The organic contaminants present in the NASA wastewater were degraded by Achromobacter sp., Rhodococcus B30 and Rhodococcus J10. While the Achromobacter sp. showed a preference for the degradation of the citric acid, the Rhodococcus species were most effective in reducing the methylhydrazine and the reaction product. Removals of more than 50% were observed for citric acid, methylhydrazine and the reaction product when the NASA wastewater was inoculated with the microbes in batch cultures. Simulation and chemical characterization of citric acid and hydrazine mixtures show that the interaction is partly of a chemical nature and leads to the formation of a conjugated UV/Visible absorbing compound. An 'azo' carbonyl derivative of the citric acid, consistent with the spectral data obtained from the investigation, has been proposed as the possible product. PMID- 10466200 TI - Population dynamics of an introduced bacterium degrading chlorinated benzenes in a soil column and in sewage sludge. AB - The capacity of the beta-Proteobacterium Pseudomonas sp. strain P51, which degrades chlorinated benzenes, to metabolize 1,2,4-trichlorobenzene (TCB) under environmental conditions was tested by its release into two experimental systems. The first system consisted of laboratory scale microcosms which were operated with and without the addition of TCB and which were inoculated with sludge from a wastewater treatment plant. The second system consisted of a non sterile, water saturated soil column. We determined survival of strain P51 after its introduction and its ability to degrade TCB. The population dynamics was followed by selective plating and applying the polymerase chain reaction (PCR) to detect strain P51 and the chlorobenzene (tcb) genes on catabolic plasmid pP51. The results showed a completely different behaviour of strain P51 in the two habitats under the applied conditions. In the soil column the P51 bacteria inoculated the entire area and their population reached 2 x 10(6) cells/g soil. The population remained active since TCB was degraded to concentrations below the detection limit of 30 micrograms/l. In the sludge microcosms, the number of strain P51 cells immediately decreased from 4 x 10(7) cells/ml to 10(5) cells/ml over a period of 2 days after inoculation, and then the strain disappeared to levels below our detection limit (10(3)-10(4) cells/ml). In the reactor without TCB the population of P51 maintained a stable value of 10(5) cells/ml during 8 days but then also decreased to levels below the detection limit. In addition, no significant TCB degradation was found in the sludge reactors. The influence of presence of TCB on maintenance of strain P51 in the two habitats is discussed. This work demonstrates the possibility to successfully apply preselected strains to degrade otherwise poorly degradable substances in complex mixed microbial communities. However, survival and activity may depend strongly on the type of system into which the strain is introduced. PMID- 10466201 TI - Biodegradation of dicyclopentadiene in the field. AB - Dicyclopentadiene (DCPD) is formed during the pyrolysis of alkanes to produce olefins suitable for manufacturing synthetic polymers. DCPD has an irritating odor with a 5 ppb detection level that provides the impetus for remediation efforts. One method of destroying odors is to alter the structure of the chemical. This can be accomplished by biological oxidation using microorganisms. Field studies at two sites, where DCPD was a soil contaminant, indicated that biodegradation contributed significantly to DCPD removal. DCPD degradation was stimulated by decreasing bulk soil density and adding nitrogen and phosphorous nutrients. The presence of other easier degradable aromatic hydrocarbons may also be beneficial, suggesting that the process is cometabolic. PMID- 10466202 TI - Analyses of polycyclic aromatic hydrocarbon-degrading bacteria isolated from contaminated soils. AB - Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria isolated from PAH contaminated soils were analyzed genotypically and phenotypically for their capacity for metabolism of naphthalene and other PAH substrates. The methods used for the analyses were DNA hybridization using NAH7-derived gene probes, PAH spray plate assays, 14C-PAH mineralization assays, and dioxygenase activity assays. The results of the analyses showed a dominant number of PAH-degrading bacteria with a NAH7-like genotype. The results support the continued use of the nahA probe for contaminated soils to monitor the genetic potential of indigenous microorganisms to degrade PAHs. However, the finding of non-nahA-hybridizing PAH-degrading bacteria show the limitation of NAH7-derived gene probes. Fifteen percent (13/89) of PAH-degrading bacteria isolated were not detected with the nahA gene probe. Four isolates (designated A5PH1, A8AN3, B1PH2, and B10AN1) did not hybridize with any of the NAH7-derived gene probes (nahA, nahG, nahH, and nahR) used in this study. Considering the numerous unculturable microorganisms in nature and their potential genotypes, NAH7-derived gene probes may underestimate the microbial potential to catabolize PAHs. This necessitates development of new gene probes for enumeration and isolation of PAH-degrading bacteria to better understand the in situ microbial potential to degrade PAHs. PMID- 10466203 TI - Carbohydrate determinants of Rhizobium-legume symbioses. AB - Rhizobium is a genus of symbiotic nitrogen-fixing soil bacteria that induces the formation of root nodules on leguminous plants and, as such, has been the subject of considerable research attention. Much of this work was initiated in response to the question 'how does recognition occur between free living rhizobial bacteria in the soil and potential host legumes?' The answer to this question has been shown to involve both cell-surface carbohydrates on the external face of the bacteria and secreted extracellular signal oligosaccharides. This review will focus on the structure, function, and biosynthesis of two of these components- the host-specific nodule-promoting signals known as Nod(ulation) factors and the rhizobial lipopolysaccharides. PMID- 10466204 TI - Synthesis of a sialyl-alpha-(2-->6)-lactosamine trisaccharide with a 5-amino-3 oxapentyl spacer group at C-1I. AB - As part of a continuing study aimed to achieve improved monoclonal antibodies against carcinoembryonic antigen (CEA) carbohydrate fragments, the synthesis of a sialyl-(2-->6)-lactosamine trisaccharide with a 5-amino-3-oxapentyl spacer group at C-1I has been developed. Two different routes to access this target are described. For this purpose 5-azido-3-oxapentyl 6-O-benzyl-2-deoxy-2-phthalimido beta-D-glucopyranoside (4) was selectively beta-galactosylated in 81% yield using the crystalline 2,3-di-O-acetyl-4,6-O-benzylidene-alpha-D-galactopyranosyl trichloroacetimidate as the donor, taking advantage of the bulky phthalimido group at C-2 of 4. On the other hand, galactosylation of the suitable protected acceptor 5-azido-3-oxapentyl 2-acetamido-3,6-di-O-benzyl-2-deoxy-beta-D glucopyranoside with the crystalline 2,3-di-O-acetyl-4,6-O-benzylidene-alpha-D galactosyl bromide renders the corresponding disaccharide in a moderate 58% yield. Despite the fact that the first strategy, unlike the second one, requires a hydrazinolysis-acetylation reaction at the disaccharide stage, it was found to be more convenient to access the disaccharide acceptor. Sialylation was performed using a thiophenyl donor under an NIS-TfOH activation procedure in acetonitrile to give a mixture of alpha and beta trisaccharides in 49 and 16% yields, respectively. PMID- 10466205 TI - Synthesis of Pseudomonas aeruginosa lipopolysaccharide core antigens containing 7 O-carbamoyl-L-glycero-alpha-D-manno-heptopyranosyl residues. AB - The monosaccharide allyl 7-O-carbamoyl-L-glycero-alpha-D-manno- heptopyranoside, the reducing disaccharide 7-O-carbamoyl-L-glycero-alpha-D- manno-heptopyranosyl (1-->3)-L-glycero-D-manno-heptopyranose and the disaccharides allyl 7-O-carbamoyl L-glycero-alpha-D-manno-heptopyranosyl-(1-->3)-L-glycero- beta- and alpha-D-manno heptopyranoside were prepared in good yields. The 7-O-carbamoyl substituent was regioselectively introduced via NH3-NH4HCO3 treatment of a 6,7-O-carbonate group. Glycosylation steps were carried out using Me3SiOTf or BF3.Et2O promoted coupling of allyl alcohol with trichloroacetimidate or fluoride glycosyl donors, respectively. The deprotected allyl glycosides were reacted with cysteamine to afford spacer glycosides which were subsequently linked to bovine serum albumin. The artificial antigens which are related to the dephosphorylated heptose region of the lipopolysaccharide core region from Pseudomonas aeruginosa classified into RNA group I may be used for the characterization of monoclonal antibodies directed against inner core epitopes of human-pathogenic Pseudomonas species. PMID- 10466206 TI - Synthesis of three diosgenyl saponins: dioscin, polyphyllin D, and balanitin 7. AB - Dioscin, polyphyllin D, and balanitin 7, which belong to a group of structurally similar diosgenyl saponins with promising bioactivities, were synthesized by stepwise glycosylation. PMID- 10466207 TI - Identification of a hexasaccharide sequence able to inhibit thrombin and suitable for 'polymerisation'. AB - Three hexasaccharides, having from low to very high affinity for antithrombin, were synthesised from disaccharide building block precursors. One of them, methyl(sodium 2,3-di-O-methyl-4-O- sodium sulfonato-alpha-L-idopyranosyluronate) (1-->4)-[(2,3,6-tri-O-sodiu m sulfonato-alpha-D-glucopyranosyl)-(1-->4)-(sodium 2,3-di-O-methyl-alpha-L-idopyranosyluronate)-(1-->4)]2-2,3,6-tri-O-sodiu m sulfonato-alpha-D-glucopyranoside, obtainable from a single disaccharide building block precursor, constitutes a good starting point for obtaining simple oligosaccharidic heparin mimetics able to inhibit the two coagulation factors thrombin and Factor Xa. PMID- 10466208 TI - Assessment through chemical synthesis of the size of the heparin sequence involved in thrombin inhibition. AB - Deca- to eicosasaccharides having the generic structure methyl(sodium 2,3-di-O methyl-4-O-sodium sulfonato-alpha-L-idopyranosyluronate)-(1-->4)-[(2,3,6-tri-O sodiu m sulfonato-alpha-D-glucopyranosyl)-(1-->4)-(sodium 2,3-di-O-methyl-alpha-L idopyranosyluronate)-(1-->4)]n-2,3,6-tri-O-sodiu m sulfonato-alpha-D glucopyranoside have been synthesized from a single disaccharide precursor. All of them bind to and activate antithrombin. When n < or = 6 only Factor Xa inhibition is observed, whereas when n > 6 Factor Xa and thrombin are both inhibited in the presence of antithrombin. These results indicate that, in heparin, the sequence involved in antithrombin-catalyzed thrombin inhibition is a pentadeca- or a hexadecasaccharide. PMID- 10466210 TI - Synthesis of methyl beta-D-arabinofuranoside 5-[1D (and L)-myo-inositol 1 phosphate], the capping motif of the lipoarabinomannan of Mycobacterium smegmatis. AB - The total synthesis of methyl beta-D-arabinofuranoside 5-(myo-inositol 1 phosphate), the capping motif of the lipoarabinomannan (LAM) of Mycobacterium smegmatis, has been completed. The stereoselective synthesis of beta-D arabinofuranosides has been achieved via an internal aglycon delivery approach using Ogawa and Ito's method. Coupling with enantiomeric myo-inositol derivatives gave the diastereoisomeric title compounds in good overall yield. Comparison with the natural product firmly established the proposed structure for the capping of the LAM but left the absolute configuration of the myo-inosityl moiety undetermined. PMID- 10466209 TI - Inhibition of some hepatic lysosomal glycosidases by ethanolamines and phenyl 6 deoxy-6-(morpholin-4-yl)-beta-D-glucopyranoside. AB - The hepatic lysosomal glycosidases alpha-glucosidase and beta-glucuronidase were inhibited in vitro and in vivo by mono- and diethanolamines. The in vivo inhibition is dose dependent and occurs at a value less than LD50. Phenyl 6-deoxy 6-(morpholin-4-yl)-beta-D-glucopyranoside inhibited alpha-glucosidase both in vitro and in vivo. The treatment of the enzymes in vitro by ethanolamine exhibited a reversible inhibition of the mixed and competitive types for alpha glucosidase and beta-glucuronidase, respectively. Diethanolamine showed a reversible inhibition of the competitive type for both enzymes. It is a potent inhibitor for beta-glucuronidase, in vitro, whose inhibition constant (Ki) is 5 x 10(-5) M. Phenyl 6-deoxy-6-(morpholin-4-yl)-beta-D-glucopyranoside is a potent inhibitor only for hepatic alpha-glucosidase with a Ki value of 1.6 x 10(-5) M. The pattern of the pH dependence of enzymic activity was not affected by ethanolamine inhibition. The magnitude of the inhibition of enzymes is dependent on the structure of the inhibitor. PMID- 10466211 TI - Endoglucanase V and a phosphatase from Trichoderma viride are able to act on modified exopolysaccharide from Lactococcus lactis subsp. cremoris B40. AB - EPS B40 from Lactococcus lactis subsp. cremoris consists of a repeating unit of- >4)-beta-D-Glcp-(1-->4)-[alpha-L-Rhap-(1 -->2)][alpha-D-Galp-1-PO4-3]-beta-D-Galp (1-->4)-beta-D-Glcp-(1-->. A phosphatase from Trichoderma viride was able to release phosphate, but only after removal of rhamnosyl and galactosyl residues by mild CF3CO2H treatment. Purified endoV from T. viride was able to act on the backbone of the polymer, but only if rhamnosyl substituents and phosphate had been removed. After complete removal of phosphate and partial removal of rhamnosyl residues by HF treatment, incubation with endoV resulted in a homologous series of oligomers. Purification of these oligomers and subsequent characterisation by NMR demonstrated that endoV was able to cleave the beta-(1- >4) linkage between two glucopyranosyl residues when the galactopyranosyl residue towards the nonreducing end is unsubstituted. The mode of action of endoV on HF treated EPS B40 is discussed on the basis of the subsite model described for endoV [J.-P. Vincken, G. Beldman, A.G.J. Voragen, Carbohydr. Res., 298 (1997) 299 310]. PMID- 10466212 TI - Structural characterisation of lipo-chitin oligosaccharides isolated from Bradyrhizobium aspalati, microsymbionts of commercially important South African legumes. AB - The shoots of the South African legume Aspalathus linearis spp. linearis (A. linearis) are used in the manufacture of an increasingly popular beverage that has acclaimed beneficial effects on health; this important export product is known as Rooibos (or Redbush) tea. Three strains of Bradyrhizobium aspalati, which are the nitrogen-fixing symbionts of Aspalathus carnosa, A. hispida and A. linearis, were tested for the production of lipo-chitin oligosaccharide signal molecules using thin-layer chromatographic analysis after induction with different inducers, including Rooibos tea extract, and radioactive labelling. Large-scale separation, using high-performance liquid chromatography, of lipo chitin oligosaccharides from B. aspalati isolated from A. carnosa was performed for structural characterisation using fast-atom bombardment mass spectrometry and chemical modifications followed by gas chromatography-mass spectrometric analysis. The strain was shown to secrete a family of unusual lipo-chitin oligosaccharides that are highly substituted on the nonreducing-terminal residue but unsubstituted on the reducing-terminal residue. They have a backbone of three to five beta-(1-->4)-linked N-acetyl-D-glucosamine residues substituted on the nonreducing terminus with a C16:0, C16:1, C18:0, C18:1, C19:1cy, or C20:1 fatty acyl chain, and are both N-methylated and 4,6-dicarbamoylated. PMID- 10466213 TI - Ethambutol-sugar hybrids as potential inhibitors of mycobacterial cell-wall biosynthesis. AB - Ethambutol is an established front-line agent for the treatment of tuberculosis, and is also active against Mycobacterium avium infection. However, this agent exhibits toxicity, and is considered to have low potency. The action of ethambutol on the mycobacterial cell wall, particularly the arabinan, and comparison of the structure of ethambutol with several of the cell-wall saccharides, suggested that ethambutol-saccharide hybrids might lead to agents with a more selective mechanism of action. To this end, eight ethambutol saccharide hybrids were synthesized and screened against M. tuberculosis and several clinical isolates of M. avium. PMID- 10466214 TI - Chemoenzymatic synthesis of spacer-linked oligosaccharides for the preparation of neoglycoproteins. AB - In the present work, the combination of chemical and enzymatic methods to obtain neoglycoproteins is described. Three bovine serum albumin (BSA)-conjugates, BSA [GalNAc alpha-], BSA-[Gal(beta 1-3)GalNAc(alpha-], and BSA-[Neu5Ac(alpha 2 3)Gal(beta 1-3)GalNAc(alpha-], were prepared. alpha GalNAc derivatives were galactosylated employing crude beta-galactosidase from bovine testes. The use of oversaturated donor solutions (pNP beta Gal) enhanced the yields up to 60%. This method was verified using divalent structures as acceptors, that rendered di- and tri-galactosylated products. Further treatment of the disaccharides with CMP Neu5Ac and alpha 2-3 sialyltransferase from pork liver led to formation of trisaccharides. Finally, mono-, di-, and trisaccharides were coupled to BSA employing a thiolic group introduced into the protein for Michael addition to a maleinimide group in the spacer-arm of the saccharide components. The results were monitored by HPLC and MALDI-TOF. PMID- 10466215 TI - Synthesis of bromoindolyl 4,7-di-O-methyl-Neu5Ac: specificity toward influenza A and B viruses. AB - N-Acetylneuraminic acid (Neu5Ac) was converted into the methyl ester methyl ketoside-8,9-epoxy derivative (8). Methylation of 8 followed by deprotection gave 4,7-di-O-methyl-Neu5Ac (10). Compound 10 was converted into the corresponding methyl ester-chloroacetate derivative, which was subsequently coupled to 5-bromo indol-3-ol to give the chromogenic product (13). Deprotection of 13 gave 5-bromo indol-3-yl 4,7-di-O-methyl-Neu5Ac (5). The product 5 was specifically cleaved by sialidase from either influenza A or influenza B virus to give an indigo-blue precipitate, but was not cleaved by several bacterial or viral sialidases tested. The properties of product 5 relative to a fluorescent substrate for sialidase were also documented. PMID- 10466216 TI - Differential effects of plant species on a mite pest (Tetranychus utricae) and its predator (Phytoseiulus persimilis): implications for biological control. AB - The influence of plant species on the population dynamics of the spider mite pest, Tetranychus urticae, and its predator, Phytoseiulus persimilis, was examined as a prerequisite to effective biological control on ornamental nursery stock. Experiments have been done to investigate how the development, fecundity and movement of T. urticae, and the movement of P. persimilis were affected by plant species. A novel experimental method, which incorporates plant structure, was used to investigate the functional response of P. persimilis. Development times for T. urticae were consistent with published data and did not differ with plant species in a biologically meaningful way. Plant species was shown to have a major influence on fecundity (P < 0.001) and movement of the pest mite (P < 0.01), but no influence on the movement of the predator. The movement of both pest and predator was shown to be related to the density of the adult pest mites on the plant (P < 0.001). Plant structure affected the functional response, particularly in relation to the ability of the predator to locate prey at low densities. The impact of these findings on the effective use of biological control on ornamental nursery stock is discussed. PMID- 10466218 TI - The physiology of diving in a north-temperate and three tropical turtle species. AB - We examined changes in blood gases, plasma ions, and acid-base status during prolonged submergence (6 h) of four aquatic turtle species in aerated water at 20 degrees C. Our objective was to determine whether the temperate species, Chrysemys picta bellii, exhibits greater tolerance to submergence apnea than the tropical species, Pelomedusa subrufa, Elseya novaeguineae, and Emydura subglobosa. Blood was sampled from indwelling arterial catheters for measurements of blood PO2, PCO2, pH, and hematocrit and for plasma concentrations of lactate, glucose, Na+, K+, Cl-, total Ca, and total Mg. The pattern of change was similar in all species: a combined respiratory and metabolic acidosis associated with a marked decrease of blood PO2. The severity of the acidosis developed in the temperate species, however, was significantly less than that of the tropical turtles. Lactate rose significantly and HCO3- fell proportionately in all turtles; changes in other plasma ion concentrations were small but were generally in the directions consistent with compensatory exchanges with other body compartments; i.e., cations (K+, Ca, and Mg increased) and anions (Cl- decreased). The results indicate that hypoxia tolerance is a conserved trait in turtles, even in those that do not experience enforced winter submergence, and that the temperate species may be superior in this capacity because of reduced metabolic rate. PMID- 10466217 TI - Cardiovascular control via angiotensin II and circulating catecholamines in the spiny dogfish, Squalus acanthias. AB - The contributions of circulating angiotensin II (Ang II) and catecholamines to cardiovascular control in the spiny dogfish were investigated by monitoring the effects of exogenous and endogenous dogfish [Asn1, Pro3, Ile5]-Ang II (dfAng II) on plasma catecholamine levels and blood pressure regulation. Bolus intravenous injections of dfAng II (30-1200 pmol kg-1) elicited dose-dependent increases in plasma adrenaline and noradrenaline concentrations, caudal artery pressure (PCA), and systemic vascular resistance (RS), and a decrease in cardiac output (Q). Similar injections of Ang II in dogfish pre-treated with the alpha-adrenoceptor antagonist yohimbine (4 mg kg-1) also elicited dose-dependent increases in plasma catecholamine levels yet the cardiovascular effects were abolished. Dogfish treated with yohimbine were hypotensive and had elevated levels of plasma Ang II and catecholamines. Intravenous injection of the smooth muscle relaxant papaverine (10 mg kg-1) elicited a transient decrease in PCA and RS, and increases in plasma Ang II and catecholamine levels. In dogfish first treated with lisinopril (10(-4) mol kg-1), an angiotensin converting enzyme inhibitor, papaverine treatment caused a more prolonged and greater decrease in PCA and RS, an attenuated increase in plasma catecholamines, and no change in plasma Ang II. By itself, lisinopril treatment had little effect on PCA, and no effect on RS, plasma Ang II or catecholamines. In yohimbine-treated dogfish, papaverine treatment elicited marked decreases in PCA, RS, and Q, and increases in plasma Ang II and catecholamines. Among the three papaverine treatments, there was a positive linear relationship between plasma Ang II and catecholamine concentrations, and the cardiovascular and hormonal changes were most pronounced in the yohimbine + papaverine treatment. Therefore, under resting normotensive conditions, while Ang II does not appear to be involved in cardiovascular control, catecholamines play an important role. However, during a hypotensive stress elicited by vascular smooth muscle relaxation. Ang II indirectly contributes to cardiovascular control by dose-dependently stimulating catecholamine release. PMID- 10466219 TI - Diurnal and seasonal changes in blood composition of the free-living Egyptian fruit bat (Rousettus aegyptiacus). AB - We studied the blood profile of the free-living fruit bat (Rousettus aegyptiacus) during the beginning of the activity period (around various feeding trees) and upon return to the day roost during 1994-1995. Results of the present study suggest that during winter and early spring bats are characterized by a poor physical and physiological state as reflected in the blood profile, revealing elevated urea and uric acid concentrations. It was found that at the end of the resting phase, R. aegyptiacus was in a mild state of dehydration (increased hematocrit and hemoglobin levels). At the end of activity, upon return to the day roost, both hematocrit and hemoglobin levels decreased but bats still maintained a high plasma osmolality. Several components in the blood are effected by the feeding time and show a cyclic change in concentration. The reverse relationship between glucose and triglyceride levels may indicate that glucose is the energy source during the active phase and that fat is the energy source during the resting period. The low cholesterol level in the blood reflects its absence in the fruit diet. PMID- 10466220 TI - What role for membranes in determining the higher sodium pump molecular activity of mammals compared to ectotherms? AB - The major body organs of mammals have sodium pumps that turn over energy (ATP) three to four times faster than those of ectotherms, at the same temperature. To examine if membranes play a role in these differences in molecular activity, membrane cross-over experiments were performed using two representative species, Rattus norvegicus and Bufo marinus. Microsomal membrane of kidney and brain displayed characteristic molecular activity differences (three- to four-fold) between the species. These molecular activity differences could be removed by delipidation. Pre-existing molecular activities and differences could be restored when reconstituted with original membrane. Using the same reconstitution method, species membrane cross-over experiments resulted in toad sodium pumps in rat membrane significantly increasing (approximately 30-40%) and rat sodium pumps in toad membrane significantly decreasing (approximately 40%) activities in both kidney and brain. Analysis of membrane composition showed reduced cholesterol content and differences in the fatty acids of phospholipids with higher overall unsaturation in the mammal. The scope for membranes to determine protein performance and its broader implications for metabolism are discussed. PMID- 10466221 TI - Calcium and ATP regulation of ion transport in larval frog skin. AB - Ion transport measured as short circuit current (Isc) across the skin of larval frogs is activated by amiloride, acetylcholine, and ATP. In many epithelia, ATP stimulation of Isc involves an increase in intracellular calcium. To define the role of changes in intracellular calcium in ATP stimulation of Isc in larval frog skin, epithelial cells were loaded with calcium by adding 5 microM ionomycin to a 2 mM calcium apical Ringer's solution. Calcium loading had no observable effect on baseline Isc or on stimulation by ATP. Minimizing changes in intracellular calcium by loading the cell with the calcium chelator BAPTA also had no measurable effect on ATP stimulation of Isc. When the apical side was bathed with Ca(2+)-free Ringer's solution, ionomycin increased Isc up to 15 microA. This increase was partially blocked by 2 mM Ca2+, 2 mM Mg2+, and 10 microM W-7. Other experiments showed that baseline-stimulated and ATP-stimulated Isc were always larger in 2 mM Mg2+ Ringer's compared to 2 mM Ca2+. In dissociated cells bathed in 2 mM Ca2+ Ringer's, ATP had no effect on intracellular calcium as measured by Fluo-LR fluorescence changes. In conclusion, ATP apparently stimulates Isc without concomitant changes in intracellular calcium. This is consistent with a directly ligand-gated receptor at the apical membrane with P2X-like characteristics. PMID- 10466222 TI - Electrophysiological and ultrastructural correlates of cryoinjury in sciatic nerve of the freeze-tolerant wood frog, Rana sylvatica. AB - We investigated function and ultrastructure of sciatic nerves isolated from wood frogs (Rana sylvatica) endemic to the Northwest Territories, Canada, following freezing at -2.5 degrees C, -5.0 degrees C, or -7.5 degrees C. All frogs frozen at -2.5 degrees C, and most frogs (71%) frozen at -5.0 degrees C, recovered within 14 h after thawing began; however, frogs did not survive exposure to -7.5 degrees C. Sciatic nerves isolated from frogs frozen at -7.5 degrees C were refractory to electrical stimulation, whereas those obtained from frogs surviving exposure to -2.5 degrees C or -5.0 degrees C generally exhibited normal characteristics of compound action potentials. Frogs responded to freezing by mobilizing hepatic glycogen reserves to synthesize the cryoprotectant glucose, which increased 20-fold in the liver and 40-fold in the blood. Ultrastructural analyses of nerves harvested from frogs in each treatment group revealed that freezing at -2.5 degrees C or -5.0 degrees C had little or no effect on tissue and cellular organization, but that (lethal) exposure to -7.5 degrees C resulted in marked shrinkage of the axon, degeneration of mitochondria within the axoplasm, and extensive delamination of myelin sheaths of the surrounding Schwann cells. PMID- 10466223 TI - Methoxybifurcarenone: an antifungal and antibacterial meroditerpenoid from the brown alga Cystoseira tamariscifolia. AB - A meroditerpenoid metabolite has been isolated from the brown alga Cystoseira tamariscifolia and characterized as methoxybifurcarenone, by spectral analysis. Methoxybifurcarenone possesses antifungal activity against three tomato pathogenic fungi: Botrytis cinerea, Fusarium oxysporum sp. mycopersici and Verticillium alboatrum and antibacterial activity against Agrobacterium tumefaciens and Escherichia coli. PMID- 10466224 TI - An amide of L-threo-gamma-hydroxyglutamic acid from Justicia ghiesbreghtiana. AB - A new amide of threo-gamma-hydroxyglutamic acid, justiciamide, was isolated from Justicia ghiesbreghtiana and shown to be (-)-N-(2-hydroxy-4,5-dimethoxyphenyl)-(2 S,4 S)-gamma-hydroxyglutamic acid. Justiciamide is the first amide of an uncommon amino acid found in this genus. PMID- 10466225 TI - Quinoline alkaloids from Acronychia laurifolia. AB - Bioassay-directed fractionation of a root extract of Acronychia laurifolia (Rutaceae) using the KB-V1+ human tumor cell line led to the isolation of six quinoline alkaloids. One of these alkaloids is novel, namely, 2,3-methylenedioxy 4,7-dimethoxyquinoline and the other five were identified as the known compounds, evolitrine, gamma-fagarine, skimmianine, kokusaginine and maculosidine. Two known bis-tetrahydrofuran lignans, sesamolin and yangambin, were also identified. The structure of the new alkaloid was determined by spectroscopic methods. All of the isolates were evaluated against a panel of human cancer cell lines; four of the alkaloids showed weak cytotoxic activity. PMID- 10466226 TI - Flavonoids from Maclura tinctoria. AB - Seven flavonoids, including two new natural products, were isolated from an ethanol extract of the bark of Maclura tinctoria (L.) Gaud. The new compounds are steppogenin 4'-O-beta-D-glucoside and orobol 5,3'-di-O-methyl-8-C-glucoside. Orobol, steppogenin, aromadendrin, dihydromorin and orobol 7-O-beta-D-glucoside were also isolated. PMID- 10466227 TI - Human health implications of environmental contaminants in Arctic Canada: a review. AB - This paper assesses the impact on human health of exposure to current levels of environmental contaminants in the Canadian Arctic, and identifies the data gaps that need to be filled by future human health research and monitoring. The concept of health in indigenous groups of the Arctic includes social, cultural, and spiritual dimensions. The harvesting, sharing and consumption of traditional foods are an integral component to good health among Aboriginal people influencing both physical health and social well-being. Traditional foods are also an economic necessity in many communities. Consequently, the contamination of country food raises problems which go far beyond the usual confines of public health and cannot be resolved by health advisories or food substitutions alone. The primary exposure pathway for the contaminants considered in this paper is through the traditional northern diet. For the Inuit, the OCs of primary concern at this time from the point of view of exposure are chlordane, toxaphene, and PCBs. Exposures are higher in the eastern than in the western region of the North. For Dene/Metis, exposure to OCs is in general below a level of concern. However, estimated intake of chlordane and toxaphene has been found to be elevated for certain groups and is a cause for concern if exposures are elevated on a regular basis. The developing foetus and breast-fed infant are likely to be more sensitive to the effects of OCs and metals than individual adults and are the age groups at greatest risk in the Arctic. Extensive sampling of human tissues in the Canadian north indicate that a significant proportion of Dene, Cree and Inuit had mean maternal hair mercury levels within the 5% risk-range proposed by the WHO for neonatal neurological damage. Based on current levels, lead does not appear to pose a health threat while cadmium is likely only a major risk factor for heavy smokers or consumers of large amounts of organ meats. Consumers of traditional foods are exposed to an approximately seven-fold higher radiation dose than non-consumers of traditional foods due predominantly to the bioaccumulation of natural radionuclides in the food chain. Risk determination for contaminants in country food involves a consideration of the type and amounts of food consumed and the sociocultural, nutritional, economic, and spiritual benefits associated with country foods. Risk management options that minimize the extent to which nutritional and sociocultural aspects of Aboriginal societies are compromised must always be considered. PMID- 10466228 TI - Spatial and temporal trends and effects of contaminants in the Canadian Arctic marine ecosystem: a review. AB - Recent studies have added substantially to our knowledge of spatial and temporal trends of persistent organic pollutants and heavy metals in the Canadian Arctic marine ecosystem. This paper reviews the current state of knowledge of contaminants in marine biota in the Canadian Arctic and where possible, discusses biological effects. The geographic coverage of information on contaminants such as persistent organochlorines (OCs) (PCBs, DDT- and chlordane-related compounds, hexachlorocyclohexanes, toxaphene) and heavy metals (mercury, selenium, cadmium, lead) in tissues of marine mammal and sea birds is relatively complete. All major beluga, ringed seal and polar bear stocks along with several major sea bird colonies have been sampled and analysed for OC and heavy metal contaminants. Studies on contaminants in walrus are limited to Foxe Basin and northern Quebec stocks, while migratory harp seals have only been studied recently at one location. Contaminant measurements in bearded seal, harbour seal, bowhead whale and killer whale tissues from the Canadian Arctic are very limited or non existent. Many of the temporal trend data for contaminants in Canadian Arctic biota are confounded by changes in analytical methodology, as well as by variability due to age/size, or to dietary and population shifts. Despite this, studies of OCs in ringed seal blubber at Holman Island and in sea birds at Prince Leopold Island in Lancaster Sound show declining concentrations of PCBs and DDT related compounds from the 1970s to 1980s then a levelling off during the 1980s and early 1990s. For other OCs, such as chlordane, HCH and toxaphene, limited data for the 1980s to early 1990s suggests few significant declines in concentrations in marine mammals or sea birds. Temporal trend studies of heavy metals in ringed seals and beluga found higher mean concentrations of mercury in more recent (1993/1994) samples than in earlier collections (1981-1984 in eastern Arctic, 1972-1973 in western Arctic) for both species. Rates of accumulation of mercury are also higher in present day animals than 10-20 years ago. Cadmium concentrations in the same animals (eastern Arctic only) showed no change over a 10-year period. No temporal trend data are available for metals in sea birds or polar bears. There have been major advances in knowledge of specific biomarkers in Canadian Arctic biota over the past few years. The species with the most significant risk of exposure to PCBs and OC pesticides may be the polar bear which, based on comparison with EROD activity in other marine mammals (beluga, ringed seal), appears to have elevated CYP1A-mediated activity. The MFO enzyme data for polar bear, beluga and seals suggest that even the relatively low levels of contaminants present in Arctic animals may not be without biological effects, especially during years of poor feeding. PMID- 10466229 TI - Spatial and temporal trends of contaminants in Canadian Arctic freshwater and terrestrial ecosystems: a review. AB - The state of knowledge of contaminants in Canadian Arctic biota of the freshwater and terrestrial ecosystems has advanced enormously since the publication of the first major reviews by Lockhart et al. and Thomas et al. in The Science of the Total Environment in 1992. The most significant gains are new knowledge of spatial trends of organochlorines and heavy metal contaminants in terrestrial animals, such as caribou and mink, and in waterfowl, where no information was previously available. Spatial trends in freshwater fish have been broadened, especially in the Yukon, where contaminant measurements of, for example, organochlorines were previously non-existent. A review of contaminants data for fish from the Northwest Territories, Yukon and northern Quebec showed mercury as the one contaminant which consistently exceeds guideline limits for subsistence consumption or commercial sale. Lake trout and northern pike in the Canadian Shield lakes of the Northwest Territories and northern Quebec generally had the most elevated levels. Levels of other heavy metals were generally not elevated in fish. Toxaphene was the major organochlorine contaminant in all fish analyzed. The concentrations of organochlorine contaminants in fish appear to be a function not only of trophic level but of other aspects of the lake ecosystem. Among Arctic terrestrial mammals, PCBs and cadmium were the most prominent contaminants in the species analyzed. Relatively high levels (10-60 micrograms g-1) of cadmium were observed in kidney and liver of caribou from the Yukon, the Northwest Territories and northern Quebec, with concentrations in western herds being higher than in those from the east. For the organochlorine contaminants, a west to east increase in zigma PCBs, HCB and zigma HCH was found in caribou, probably as a result of the predominant west to east/north-east atmospheric circulation pattern which delivers these contaminants from industrialized regions of central and eastern North America to the Arctic via long-range atmospheric transport. Radiocesium contamination of lichens and caribou has continued to decrease. Significant contamination by PCBs and lead of soils and vascular plants was observed in the immediate vicinity and within a 20-km radius of DEW line sites in the Canadian Arctic. There was also evidence for transfer of PCBs from plants to lemmings. There was no evidence, however, that large mammals such as caribou living in the general area of the DEW line sites had elevated levels of PCBs. There is very limited temporal trend information for most contaminants in biota of Arctic terrestrial and freshwater environments. PMID- 10466231 TI - Eutrophication process recorded in dinoflagellate cyst assemblages--a case of Yokohama Port, Tokyo Bay, Japan. AB - To investigate temporal changes of water quality, a role of dinoflagellate cysts preserved in surface sediments was examined in Yokohama Port in Tokyo Bay, Japan. Two cores were collected, and sedimentation rates and ages of both were dated as approximately 1900 years or slightly older on the basis of 210Pb and 137Cs concentrations. The temporal change in dinoflagellate cyst assemblages in the two cores reflects eutrophication in Yokohama Port in the 1960s. Abrupt increases in the cysts of Gyrodinium instriatum cysts strongly suggests that a red tide was caused by this species around 1985. Dinoflagellate cyst assemblages in surface sediments appear to be good biomarkers of changes in the water quality of enclosed seas. PMID- 10466230 TI - Sedimentary 4-desmethyl sterols and n-alkanols in an eutrophic urban estuary, Capibaribe River, Brazil. AB - Sterols, n-alkanols, organic carbon (OC), C/N ratios and carbon isotope data (delta 13C) were investigated in sediments of the urban Capibaribe River estuary, NE Brazil, in order to assess allochthonous and autochthonous sources of organic matter (OM). Sedimentary OC values are high, but C/N ratios and delta 13C data generally fall within the range of values reported in other riverine systems, and suggest mixed inputs from aquatic and terrestrial matter. Mean values for total 4 desmethyl sterols and high molecular weight (HMW) n-alkanols are 11.0 micrograms/g and 2.8 micrograms/g, respectively. Sterols are found at highest levels in areas of enhanced urban outfalls. They can be related to major planktonic species growing in riverine waters. Stanol/stenol ratios suggest a high degree of alteration of the autochthonous OM as a result of elevated temperatures and microbiological proliferation. Even though sterols suggest the importance of autochthonous inputs to the river, HMW n-alkanols indicate major terrigenous accumulation at the mouth and 10 km upriver. Coprostanol and epicoprostanol levels are comparable to other sewage contaminated hydrosystems, but not as high as expected given the importance of sewage outfalls and low riverine water discharge. However, high (coprostanol)/(coprostanol + cholestanol) ratio values indicate that fecal contamination is significant. PMID- 10466232 TI - Radiocaesium activity concentrations in the fruit-bodies of macrofungi in Great Britain and an assessment of dietary intake habits. AB - Radiocaesium activity concentrations in the fruit-bodies of some species of macrofungi are higher than in many other foodstuffs. The consumption of fruit bodies contributes significantly to radiocaesium intake of humans in some countries. In the United Kingdom, the collection of wild fungi has generally been considered to be of minor importance and there are few data on consumption rates or radiocaesium activity concentrations in most edible species. Samples of commonly eaten species in Great Britain have been collected to assess radiocaesium contamination levels and geographical variation. Concurrently, surveys of consumption habits were conducted. A total of 425 samples representing 37 different species were collected. Significantly higher radiocaesium activity concentrations occurred in mycorrhizal compared to saprotrophic or parasitic species. The highest 137Cs activity concentration of 30.5 kBq kg-1 dry wt. was determined in a sample of Hydnum repandum collected in Wales. The transfer of radiocaesium from soil to fungal fruit-bodies was highly variable, ranging over three orders of magnitude within individual species. A number of approaches to quantifying radiocaesium transfer from soil to fungal fruit-bodies were used. Although these were in general agreement with previously measured values in other countries, all the approaches gave variable results. Over 200 people responded to the dietary habits questionnaire. The median intake rate was 0.75 kg year-1 (fresh wt.) and 60% of respondents consumed only one species (generally Agaricus campestris). However, intakes of up to 26 kg year-1 were recorded and a total of 82 species were consumed. The intake of 137Cs was determined by the amount of mycorrhizal fungi in the diet rather than the total intake of fungi. Assuming median recorded 137Cs activity concentrations in each fungal species, the estimated annual committed effective dose for over 95% of respondents was < 1 microSv. Hence, currently, the consumption of wild fungi in the UK would not be expected to significantly increase the dose above that attributable to the normal diet of most consumers. However, the results of this study demonstrate that, in the event of any future accidental release of radiocaesium, the potential ingestion dose received from the consumption of wild fungi would need to be considered. PMID- 10466233 TI - [Current trends and therapeutic strategies in breast carcinoma. Introduction]. AB - Carcinoma of the breast is a pathology of great social interest for its increase in occurrence and because it is leading cause of tumor-induced death among women in Western Countries. There have been continual changes in the understanding of biological implications of the tumor and in the ensuing therapeutic approaches. Breast cancer is currently viewed as a systemic disorder right from is outset varies widely in behavior within the same histologic type. Hence, the need for a multidisciplinary approach where locoregional treatment (surgery and radiotherapy) is accompanied by systemic (chemotherapy and hormone therapy). As to surgery, there has been a shift away from the radical to the conservative approach which avoids functional and above all psychological mutilation. Beside always being carried out when surgery is not radical, radiotherapy depends on the stage of the tumor. As to chemotherapy, while there is sound evidence proving it to be useful in postoperative adjuvant treatment, its usefulness in preoperative treatment still remains to be demonstrated. Future studies will have to identify new morphobiological parameters of the tumor capable of predicting the clinical behavior and response to therapy of the various types of breast cancer so as to correctly standardize therapeutic actions and their aiming. PMID- 10466234 TI - [Guideline concepts of the present-day surgery in breast cancer]. AB - The more important trends of nowadays surgery for breast cancer are those of an advanced rationalization of combined strategies in a multidisciplinary view. Moreover it takes advantages from the availability of an extremely modern diagnostic instrumentation used in search of a truly early diagnosis, based on scheduled screening and surveillance criteria both for general population and particularly for that at risk. In this way it is easier and possible to adopt less radical and more conservative surgical techniques, whose effectiveness is well demonstrated in association with adjuvant treatment thanks to the convergence of multiple competences. The better results of the last years are certainly related to the collaboration of surgeons with oncologists, thanks to the ever more effectiveness of adjuvant chemo- and radiotherapy, that make more acceptable the attitude for the reconstructive and the conservative surgery whenever possible. This interpretation of the actual positive results of breast cancer surgery point the way to follow, and are the basis for hoping further improvement in the treatment of this disease, ever more efficacious whilst that diagnosis precocious be more and more. PMID- 10466235 TI - [Prognostic-predictive factors and therapeutic choices in invasive carcinoma of the breast]. AB - Numerous pathologic and biologic factors have been assessed for their ability to predict clinical behaviour and response to therapy in breast cancer patients. Currently, traditional morphological features such as lymph node status, tumour size, histological type and tumour grade remain the leading prognostic factors. Steroid receptors are also important, but their main role consists in predicting response to hormonal therapy. Although several new indicators have been proposed to refine the value of the above-mentioned proven parameters, interpretation of their impact is not conclusive. In particular, immunohistochemical evaluation of p53, c-erbB-2 and MIB-1 appears to be potentially useful and considerable efforts are currently being made to technically and clinically validate them. PMID- 10466237 TI - [Axillary lymphadenectomy]. AB - Axillary lymphadenectomy represents one of the historical hallmarks in the surgical strategy for breast cancer treatment. In recent years the role of axillary dissection is becoming matter for a renewed debate: up to date better therapeutic results together with new biologic acquisitions and progresses in early diagnosis have been influencing current indications and extension of axillary lymphadenectomy; also, cosmetic and financial considerations play a relevant role. This is the background of a large number of studies investigating the possibility of a more limited applications of axillary dissection provided that oncologic results remain unchanged. In this context sentinel node biopsy is one of the most promising lines of research. It can be stated that, as far as early stages are concerned, a trend towards more conservative resection techniques that started with the first quadrantectomies two decades ago, is now involving axillary dissection too. Possibly, in the next future, an elective axillary lymphadenectomy will not be performed any more if the presence of positive nodes will not be preliminarily ascertained. Nevertheless such conservative strategy can not be recommended to date, until the technique of sentinel node biopsy will be optimized and its results confirmed by randomized trials. PMID- 10466236 TI - Ductal carcinoma in situ of the human breast: clinico-pathological aspects. AB - The apparent increase of incidence of DCIS is related to the development of mammographic screening programs and requires an effort of diagnostic, classification and treatment revision. The same natural history of the DCIS is still poorly known, and the phases of its evolution are supposed essentially on epidemiologic and statistics bases. The efforts for a classification are aimed to the search of a prognostic meaning: nuclear grading and possible invasive evolution; association of nuclear grading and necrosis. Both are correlated with local recurrence and free interval. The finding of micro-calcifications is the basis of the instrumental mammographic diagnosis; the possible isolated and unique finding encourages choices of conservative surgery. But as the less large lesions are as a rule of higher grade than those more extended, there are the treatment can be puzzling. As a consequence in fact the former are exposed to wider resections than the last ones. The DCIS is also characterized in its post surgical evolution from the borders of the excision, from the pathologic classification, from the nuclear degree and from the dimensions of the lesion. In the more favourable conditions the surgical excision represents an adequate treatment. The radiotherapy finds his role in the more elevated degrees and in the case of incomplete excision, also if only presumptive. The alternative is the reoperation and the mastectomy. PMID- 10466238 TI - [Breast reconstruction]. AB - The ideal goal of every post-mastectomy breast reconstruction is to achieve a breast that is as "identical" as possible to the contralateral one in shape, size, consistency, mobility and degree of naturalness. At the same time, however, it is essential to rely on the simplest and safest reconstruction technique as far as the patient is concerned. Mastectomy and restoration of the lost morphology should be performed in a single operation whenever the opportunity arises and there are two reasons for this. First of all, by working this way reconstruction can be performed on an area that is completely free of any scar tissue. Secondly, there are psychological considerations involved, since this makes it possible to avoid the trauma of letting the patient seeing herself, even for just an instant, without her breast. The numerous different techniques used for the morphological reconstruction of breast volume can be classified into two basic groups: reconstruction with insertion of a prosthesis and reconstruction without a prosthesis. Both of these techniques make it possible to personalize surgical choices based on the patient's unique characteristics, in order to achieve a complete symmetry in relation to the contralateral breast. PMID- 10466239 TI - [Primary chemotherapy in operable breast carcinoma]. AB - We have reviewed the current status of primary chemotherapy for resectable breast cancer. Available findings indicate that the most immediate benefit of this treatment approach consists in an important tumor shrinkage, enabling to significantly increase the rates of breast conserving approaches. In spite of the logical and scientific rationale, available data from prospective randomized studies do not provide evidence of a clear superiority of primary chemotherapy over adjuvant chemotherapy. Nevertheless, the observed kinetic acceleration of micrometastases following noncurative surgical excision in animal studies represents a strong biologic evidence supporting primary chemotherapy. From a clinical perspective, primary chemotherapy with conventional regimens can induce a low to moderate rate of pathological complete remissions that appear to be an important marker of favorable treatment outcome. For this reason, well designed, prospectively randomized, large trials with drug regimens more effective than those used so far are mandatory to demonstrate the real worthiness of this newer treatment approach and to define the optimal timing of surgery and other locoregional modalities in respect to the long-term treatment outcome. Should there be no benefit to the use of primary drug treatment, then the biologic consequences of primary tumor removal, as observed in experimental models, are not likely to be of clinical significance. Nonetheless, since primary chemotherapy can achieve long-term results similar to those obtained with postoperative drug treatment while increasing the frequency of breast-sparing approaches, women can be offered a choice to maintain their body integrity. Should improved pathological complete remission rates from more effective regimens of primary chemotherapy translate into improved survival compared with the classical postoperative modality, then, regardless of the tumor diameter at diagnosis, our overall strategy for the management of operable breast cancer will require a radical departure from the traditional dogma. PMID- 10466240 TI - [Timing of radiotherapy in the treatment of breast carcinoma. Review of the literature]. AB - Combined modality treatments are indicated for most patients with breast cancer. The definition of a proper treatment schedule and of the timing of each modality is a relevant issue that affects the feasibility and the clinical outcome of the treatment. A review of the literature was done on the timing of radiation therapy (RT) in the post-operative treatment of breast cancer. Retrospective studies and randomized clinical trials addressing the issue were considered and grouped according to the combined modality treatments performed. With regard to breast conserving surgery and adjuvant RT, it was verified that a delay up to 8 weeks between breast surgery and start of RT is not associated with an increased risk of local failure if compared with RT started within 4 weeks. Concerning breast conserving surgery followed by adjuvant RT and chemotherapy, the choice of the best schedule is still a complex unresolved issue. More results on sequential schedule of adjuvant RT and chemotherapy from recently published randomized studies are available. The reconstruction of the breast after conservative surgery is rarely necessary and is usually performed immediately after surgical treatment. With regard to mastectomy followed by adjuvant RT and breast reconstruction, it appears from some retrospective studies that autologous tissue transfer offers better results over implants. Concerning the transverse rectus abdominis muscle (TRAM) procedure, either immediate reconstruction before RT or delayed reconstruction after RT was feasible without significative differences in complication rates. Further randomized clinical studies are required to address the unresolved issues linked to the timing of RT. At present the treatment plan should be based on the patient's individual circumstances with regard to risk of metastasis and local failure. PMID- 10466241 TI - [Treatment of metastatic breast carcinoma]. AB - In spite of major advances in early detection and adjuvant therapy, metastatic breast cancer remains a major clinical problem affecting a significant number of patients. Metastatic disease is generally considered incurable and conventional therapy is used mainly with palliative intent. Therefore, the choice of appropriate therapeutic approach requires a reasoned evaluation of the likelihood of benefit from therapy balanced with the impact of therapy on the patient's quality of life. The estimated aggressiveness of the tumour and indicators of response to therapy (e.g. the status of hormonal receptors) are useful parameters to take into account before selecting a given treatment. Endocrine therapy is an important option in the management of stage IV disease, with tamoxifen the most widely used first-line drug in postmenopausal women. For progressing patients, the third generation of aromatase inhibitors (letrozole, anastrazole) are considered good second-line endocrine agents. The LH-RH agonists represent the initial choice in premenopausal patients candidate to receive hormonal therapy, with the use of tamoxifen as a valuable alternative. Combination chemotherapy regimens (e.g. CMF or CAF) are usually used for first-line treatment of patients with aggressive, steroid receptor-negative disease. Recently, several relatively new agents have shown significant activity in metastatic breast cancer. In particular, the taxanes (paclitaxel and docetaxel) are particularly promising as single agents but also in combination with other drugs, especially anthracyclines. Finally, another class of agents, the d mention because they represent an important new treatment modality in the management of metastatic. PMID- 10466242 TI - [Total thyroidectomy with identification of parathyroid glands. Functional implications]. AB - Several factors have been involved in the pathogenesis of postoperative hypocalcemia after total thyroidectomy (TT). The real cause is yet unclear, but postoperative (p.o), hypoparathyroidism seem s to be the most important factor. MATERIAL AND METHOD: 337 patients underwent TT; a systematic and accurate identification and preservation of parathyroid glands was always performed. In all patients calcemia was evaluated before and after surgery (1St, 2nd, 4th, 15th, and 30th day), moreover in 90 patients was also evaluated phosphorus, magnesium, alkaline phosphatase, total proteins, PTH, calcitonin serum levels and urinary levels of calcium and phosphorus. RESULTS: No permanent hypocalcemia was observed and transient hypocalcemia was present in 13.6% of patients. Among 90 patients, 84 showed normal calcium serum levels like the others parameters; the other 6 showed a post-operative hypocalcemia associated with clinical symptoms, an increase of phosphoremia and a decrease of PTH and phosphaturia in early p.o days; in these patients calcemia and PTH levels reached normal values within 30 days after surgery. CONCLUSION: The surgical manipulation of parathyroid glands should be the cause of lowering of PTH serum concentration and transient hypocalcemia. PMID- 10466243 TI - [Primary hyperparathyroidism. Critical analysis of a 20-year experience]. AB - PURPOSE: The aim of this study was to review our 20-year experience in the treatment of primary hyperparathyroidism (HPT) and to analyze clinical and biochemical findings in patients surgically treated in the nineties and before. MATERIAL AND METHODS: From 1978 to 1997, 302 patients with proved HPT underwent primary surgery. There were 95 (31.5%) males and 207 (68.5%) females with a median age of 53 years (range 11-82 years). Two groups of patients were considered: Group 1 (156 patients undergoing surgery between 1978 and 1989) and Group 2 (146 patients undergoing surgery from 1990 to 1997). RESULTS: No differences (p = NS) were found regarding age at operation and preoperative serum creatinine values. The most frequent symptoms in both groups were of rheumatological nature (54.5% and 42.5% in Groups 1 and 2 respectively). Renal stones were described in 60.3% and 42.5% of Group 1 and 2 patients, while peptic ulcerations or gastritis were present in 25.0% and 15.1%. No differences (p = NS) in the number of patients with neurological and neuropsychiatric rate and in the median size of the removed parathyroid glands were observed between the two groups. Asymptomatic HPT was more frequent (20.5% vs 7.7%) in patients who underwent parathyroidectomy from 1990. Average serum calcium preoperative levels and delayed diagnosis were greater (p < 0.01) in Group 1 (3.10 vs 2.90 mmol/L and 72 vs 24 months respectively). CONCLUSION: Increase of asymptomatic patients with HPT allowing an earlier surgical treatment may reduce the time of the of neck exploration and morbidity rate. PMID- 10466244 TI - [Gastric carcinomas of the "bare area". Their anatomo-surgical definition and proposal of an en bloc total gastrectomy]. AB - INTRODUCTION: Regarding the surgical treatment, carcinomas of the gastric proximal third and cardia are considered as an uniform problem. Particularly no difference exists between the gastric tumors of the anterior wall and those of the posterior wall. Nevertheless, cancers of the posterior wall of the proximal third are more difficult to cure and therefore less success has been achieved than with those of the anterior wall. The cause of this different behaviour must consist in an anatomical characteristic, the fact that the posterior wall of the fundus and subcardial portion is not covered by the visceral peritoneum ("bare area"). MATERIAL AND METHODS: Our study group consisted of 22 patients who presented carcinoma of the stomach risen in the area without peritoneal lining. Our treatment of this condition consisted of an operational procedure wherein we removed en bloc the stomach, left pancreas, spleen, and lymph nodes of the compartment I, II and III. In each case we considered the pathological situation, the involvement of the retroperitoneal structures and the condition of the lymph nodes in the compartment III specially, and actuarial statistics. RESULTS: We divided the 22 patients in 4 groups based on the neoplastic involvement of the retroperitoneum. Only in 3 cases (14%) of group 1 the cancer invasion was limited to the gastric muscular layer. In all of the rest, tumor involved the retroperitoneal structures. Postoperative mortality rate was 9%. The five year survival rate was 100% in the 3 cases of group 1 only and 10.5% in all other cases. DISCUSSION: The results show that carcinomas of the "bare area" cause a real jump in the degree of severity of the biological neoplastic process and hence of the prognostic evaluation. When the growth involves the gastric muscular layer, it indicates invasion of the retroperitoneum. Therefore, as with surgical treatment of esophageal carcinoma with the en bloc esophagectomy, we propose an en bloc total gastrectomy which should be performed in every carcinoma arising in the "bare area" because it allows more radical removal than can be obtained from traditional surgical procedures, without risking an increased mortality rate. PMID- 10466245 TI - [Lesions of the main bile duct: combined percutaneous-endoscopic treatment]. AB - The authors report their experience in "minimally invasive" treatment of benign biliary lesions. More than 80% of them occur after injury to the bile ducts during cholecystectomy. The clinical experience reported in this work concerns three patients managed with a coordinated--percutaneous and endoscopic--approach. In all patients an ERCP with biliary sphincterectomy has been performed at first, followed by percutaneous biliary drainage with external-internal catheters. All drainage catheters have been removed after one year. No patient has evidence of recurrence after a mean follow up of 18 months. Only in a patient a recurrent cholangitis treated with antibiotic therapy has been observed. This preliminary study suggests that the "rendez-vous" technique may be useful for the initial management of biliary strictures in most patients. This association improves the success rate and can avoid surgery. The role of surgery is reserved to few selected cases and particularly to lesions which are detected at the time of the operation. PMID- 10466246 TI - [Surgical therapy of right colon cancer]. AB - The debate about the management of frequent advanced right colon cancer is still opened: the opportunity of extended resections when the surrounding organs or tissues are infiltrated, the lymphadenectomy extension and its role, the possibility of identifying prognostic factors that could be useful to decide adjuvant therapy, the definition of the role of laparoscopy. Considering these problems, we have reviewed a series of 159 operations performed by the Institute of Clinical Surgery of the University of Trieste from 1980. 112 of these operations had a curative goal. The reconstruction of intestinal continuity was carried out manually in 28 cases and with mechanical stapler in 78. As far as the curative resection are concerned, in 89 of them an extended lymphadenectomy was performed (D3), while in 18 cases the lymphadenectomy was limited to the lymph nodes of first and second level due to the general bed conditions of the patient. 27 of the curative exeresis were performed in patients with T4 tumor infiltrating the nearby tissues. Referring to Dukes' classification, 8 were included in stage A, 59 in stage B and 40 in stage C, while as far as the depth of wall infiltration is concerned 2 were categorized as T1, 9 as T2, 69 as T3 and 27 as T4. The overall operative mortality was of 5 patients, the overall morbidity of 14%, that specific of 4.6%. The final incidence of local recurrences was 13.8% for Dukes A cancers, 10.9% for Dukes B and 120.5% for Dukes C (p = 0.0614). Half of the recurrences (50%) occurred in patients with a cancer infiltrating the nearby tissues. The 5 year survival rate for patients with Dukes A lesions was 100%, for patients with Dukes B lesions 73.4% and for Dukes C 52.3% (p = 0.00510). With Cox' multivariate analysis only the stage disease, T and grading showed a significative correlation with the survival rate. Our experience, therefore, suggests the execution of an exeresis with lymphadenectomy D3 in each cases where the local site of the lesion and the general conditions of the patients allow it and an extended exeresis where possible from a technical point of view and when the lymph nodes are involved. PMID- 10466248 TI - [Vascular cysts of the adrenals. Association with aneurysm of the abdominal aorta]. AB - A rare case of an adrenal vascular cyst associated to an abdominal aorta aneurysm is reported. Adrenal cysts are an uncommon clinical finding, in most cases incidentally discovered for nonspecific abdominal pain, during US, TC or RM evaluation or at autopsy. Small adrenal mass are clinically silent. They may be symptomatic (lumbar tension, pain) for dimensions over 10 centimetres. Cysts of large size can cause displacement and compression of adjacent organs. They present a difficult problem of differentiation between benign and malignant lesions. Non-neoplastic adrenal cysts have been divided into four categories: parasitic (7%), epithelial (9%), endothelial (45%) and haemorrhagic or pseudocystic (39%). Vascular adrenal cysts may be a traumatic consequence of an hamartomatous vascular anomaly. The aim of this paper is to discuss, on the basis of the literature, the etiology, diagnosis and treatment of the adrenal mass. Surgical timing is discussed for the concomitant vascular lesion. The elective treatment was left adrenalectomy performed through transperitoneal approach. Surgery for abdominal aorta aneurysm was differed because the adrenal mass was suspected to be an infected neoplastic lesion and for the feasibility of endovascular procedure. The adrenal specimens contained a cystic structure with fluid blood, fibrin and calcifications. Normal adrenal cortical tissue was found in the cystic wall. This lesion (arising from vascular anomalies) require separation from haemorrhagic adrenal neoplasm. Awareness of adrenal pseudocysts and careful attention to the hystological features aids this distinction. PMID- 10466249 TI - [Rare primary localization of echinococcus cysts. Presentation of 2 cases and review of the literature]. AB - Hydatidosis is a parasitic disease. It's generally localized in the liver and in the lung even if any other organ can be potentially affected. From 92 to 98 our group observed two cases of rare primitive localisation of echinococcosis (one mediastinal and one retroperitoneal cyst). The patients performed serologic and instrumental exams to establish the right therapeutic strategy. Both of them were operated (it was carried out a pericystectomy with open cyst in one of them and an exeresis of the right adrenal gland including hydatid cyst in the other one). A patient underwent to Albendazole prophylaxis. During the follow-up the patients performed echography, CT and MRI. Considering the literature's data and their experience the authors emphasize: 1. The diagnosis of rare primitive localization of hydatidosis is very difficult. 2. The diagnostic iter requires a correct relationship between the laboratory and imaging data. 3. Only the exclusion of other localizations of the cyst (liver, lung or other organs) give us the possibility to diagnose a rare primitive localization of hydatidosis. 4. The treatment of hydatidosis is specifically surgical. 5. The surgeon can use several different approaches in relationship to the place and the anatomopathologic characters of the cyst. PMID- 10466247 TI - [Conservative surgery in the treatment of rectal tumors. Our experience, 1986 1997]. AB - OBJECTIVE: Our institution's experience with low anterior resection for primary rectal cancer was reviewed to determine cancer treatment results and to identify risk factors for pelvic recurrence. METHODS: Ninety-two patients treated for primary rectal cancer between 1986 and 1997 were studied retrospectively. All cases was classified according to Astler-Coller. Median follow up was 46 months. RESULTS: Actuarial 5-year survival for all patients was 79%. Among 28 patients who relapsed, only 9 pelvic recurrence was detected with an actuarial rate of 15% at 5 years. Lymph nodes involvement were associated with increased risk for pelvic recurrence. DISCUSSION: Low anterior resection permitted by circular stapler provides good treatment for mid-rectal cancers and for some distal rectal cancers. Pelvic recurrence is not correlated with neoplasm localization, T stage and short distal resection margins, but only with the presence of lymph nodes involvement. This result increase the value of the hypothesis that search the pelvic recurrence origin in a extra-parietal disease respect anastomotic disease. CONCLUSION: Fundamental to prevent pelvic recurrence is the surgical technique that have to remove lymph node en bloc with the rectum doing a dissection along the endopelvic fascia to Denonvillers fascia so to take away completely the mesorectum. PMID- 10466251 TI - The naturally occurring furanones: formation and function from pheromone to food. AB - Three closely related 4-hydroxy-3(2H)-furanones have been found in a range of highly cooked foodstuffs where they are important flavour compounds with aroma threshold values as low as 20 micrograms kg-1 water (approximately 0.14 mumol l 1). The compounds are formed mainly as a result of the operation of the Maillard reactions between sugars and amino acids during heating but one compound, 5-(or 2)-ethyl-2-(or 5)-methyl-4-hydroxy-3(2H)-furanone, appears in practice to be produced by yeast, probably from a Maillard intermediate, during the fermentation stages in the production of soy sauce and beer. The compounds are also important in the flavour of strawberry, raspberry, pineapple and tomato but the route of biosynthesis is unknown. Two 3-hydroxy-2(5H)-furanones, emoxyfuranone and sotolon, which are produced spontaneously from amino acids such as threonine and 4-hydroxy-L-leucine are major contributors to meaty and spicy/nutty flavours in foods. The biosynthesis of 5-(1,2-dihydroxyethyl)-3,4-dihydroxy-2(5H)-furanone (ascorbic acid, vitamin C) and 5-hydroxymethyl-3,4-dihydroxy-2(5H)-furanone (erythroascorbic acid) from sugars in plants and yeast, respectively, has been characterized to the enzymic level. After treatment with chlorine, humic waters contain a range of chloro-furanones, some of which, particularly 3-chloro-4 (dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), are powerful mutagens. The furanones which occur in foods are also mutagenic to bacteria and cause DNA damage in laboratory tests. However, these compounds are, in practice, very effective anti-carcinogenic agents in the diets of animals which are being treated with known cancer-inducing compounds such as benzo[alpha]pyrene or azoxymethane. Two of the food-derived furanones have antioxidant activity comparable to that of ascorbic acid. A biological function has been discovered for some of the furanones besides vitamin C. 5-Methyl-4-hydroxy-3(2H)-furanone is a male pheromone in the cockroach Eurycolis florionda (Walker) and the 2,5 dimethyl derivative deters fungal growth on strawberries and is an important component of the attractive aroma of the fruit. The red seaweed Delisea pulchra (Greville) Montagne produces a range of brominated furanones which prevent colonisation of the plant by bacteria by interfering with the acylated homoserine lactone (AHL) signalling system used by the bacteria for quorum sensing. In addition, these compounds can deter grazing by marine herbivores. It is proposed here that the evolved biological function of a number of furanones is to act as inter-organism signal molecules in several different systems. This has resulted in two coincidental effects which are important for humans. Firstly, the easily oxidized nature of the furanones in general, which is likely to be an important property in their functioning as signal molecules, results in both mutagenic and anti-carcinogenic activity. The balance of these two effects from compounds in the diet has yet to be fully established. Secondly, and more specifically, the 4 hydroxy-3(2H)-furanones associated with fruit aromas act to attract animals to the fruit, which ensures seed dispersal. In the case of humans, the coincidental synthesis of some of these compounds in foods during preparation results in these foods appearing particularly attractive through the transferred operation of the original signalling mechanisms. PMID- 10466253 TI - Ultraviolet radiation screening compounds. AB - Amongst the diversity of methods used by organisms to reduce damage caused by ultraviolet (UV) radiation, the synthesis of UV-screening compounds is almost ubiquitous. UV-screening compounds provide a passive method for the reduction of UV-induced damage and they are widely distributed across the microbial, plant and animal kingdoms. They share some common chemical features. It is likely that on early earth strong selection pressures existed for the evolution of UV-screening compounds. Many of these compounds probably had other physiological roles, later being selected for the efficacy of UV screening. The diversity in physiological functions is one of the complications in studying UV-screening compounds and determining the true ecological importance of their UV-screening role. As well as providing protection against ambient UV radiation, species with effective screening may also be at an advantage during natural ozone depletion events. In this review the characteristics of a wide diversity of UV-screening compounds are discussed and evolutionary questions are explored. As research into the range of UV-screening compounds represented in the biosphere continues, so it is likely that the properties of many more compounds will be elucidated. These compounds, as well as providing us with insights into natural responses to UV radiation, may also have implications for the development of artificial UV-screening methods to reduce human exposure to UV radiation. PMID- 10466254 TI - The ethological analysis of imitation. AB - Theorists and experimental researchers have long debated whether animals are able to imitate. A variety of definitions of imitation have been proposed to describe this complex form of social learning. Experimental research on imitation has often been hampered by either a too loose 'anthropomorphic' approach or by too narrow 'behaviourist' definitions. At present neither associative nor cognitive theories are able to offer an exhaustive explanation of imitation in animals. An ethological approach to imitation offers a different perspective. By integrating questions on function, mechanism, development and evolution one can identify possible directions for future research. At present, however, we are still far from developing a comprehensive theory of imitation. A functional approach to imitation shows that, despite some evidence for imitative learning in food processing in apes, such learning has not been shown to be involved in the social transmission of either tool-use skills or communicative signals. Recently developed procedures offer possible ways of clarifying the role of imitation in tool use and visual communication. The role of imitation in explorative play in apes is also investigated and the available data suggest that copying during play might represent a behavioural homologue of human imitation. It is proposed that the ability to copy the behaviour of a companion is under a strong genetic influence in many social species. Many important factors have not been examined experimentally, e.g. the effect of the demonstrator, the influence of attention and memory and the ability to generalize. The potential importance of reinforcement raises the possibility that copying abilities serving divergent functions might be partly under the control of different mechanisms. PMID- 10466256 TI - Need for vaccination against yellow fever. PMID- 10466255 TI - Salmonella typhimurium definitive type 104 isolates in British Columbia, 1997 1998. PMID- 10466257 TI - Influenza immunisation and health care workers and other occupational groups. PMID- 10466258 TI - Cryptosporidium in water and the immunocompromised. PMID- 10466259 TI - National Scientific Medical Meeting. 26-27 March 1999. Abstracts. PMID- 10466260 TI - Introduction to the special section on the concept of disorder. AB - Despite the absence of a consensual definition of disorder, considerable research and clinical work is based on the categorization and diagnosis of mental disorder. This article introduces a special section of the Journal of Abnormal Psychology that expands the debate between J. C. Wakefield (1999), who has proposed a "harmful dysfunction" analysis of disorder and S. O. Lilienfeld and L. Marino (1995, 1999), who offer an alternative "Roschian" or prototype analysis. This introduction summarizes the main arguments of Wakefield's target article and 8 critiques and discusses the conceptual value of the debate, especially an integration of diverse viewpoints and stimulation to further consideration of this important topic. PMID- 10466261 TI - Evolutionary versus prototype analyses of the concept of disorder. AB - The harmful dysfunction (HD) analysis of the concept of disorder (J. C. Wakefield, 1992a) holds that disorders are harmful failures of internal mechanisms to perform their naturally selected functions. S. O. Lilienfeld and L. Marino (1995) proposed instead that disorder is a Roschian prototype concept without defining properties. Against the HD analysis, they argued that many disorders are not failures of naturally selected functions because they are either designed reactions (e.g., fever) or failures of functions that are not naturally selected (e.g., reading disorder). The HD analysis is defended here against these and other objections and compared with the Roschian account. It is argued that the objections are based on conceptual confusions and can be turned around to provide strong new support for the HD analysis. A series of conceptual experiments demonstrates the superior explanatory power of the HD analysis and disconfirms the Roschian account. PMID- 10466262 TI - Essentialism revisited: evolutionary theory and the concept of mental disorder. AB - J. C. Wakefield's (1999) elaboration of his harmful dysfunction analysis (HDA) of mental disorder does little to address previous criticisms (S. O. Lilienfeld & L. Marino, 1995) and instead reveals further conceptual weaknesses in his position. The authors demonstrate that (a) a Roschian analysis can account for the results of all of Wakefield's conceptual experiments and predicts a number of judgments of disorder not predicted by the HDA, (b) the HDA is incapable in many cases of providing a scientifically nonarbitrary distinction between disorder and nondisorder, and (c) the HDA cannot account for failures of cultural ex adaptations, mismatches between evolutionary design and novel environments, or defenses against threat. The authors argue that the HDA has been convincingly falsified and discuss the failure of essentialistic concepts to resolve controversies in other domains of biological science. PMID- 10466263 TI - Nine variations and a coda on the theme of an evolutionary definition of dysfunction. AB - Nine variations on the theme of J. C. Wakefield's (1999) evolutionary definition of dysfunction show that the concept is not, as he claims, purely causal. It depends also on a teleological element of meaning introduced, in Wakefield's formulation, through an equivocation on the sense in which natural selection explains biological forms. The corollary (presented here briefly as a coda to the theme and variations) is that Wakefield's definition is not, as he also claims, value free. However, contra S. O. Lilienfeld and L. Marino (1995), this does not place diagnostic judgments of dysfunction outside the scope of science. PMID- 10466264 TI - Harmful dysfunction, disorder, disease, illness, and evolution. AB - Can psychiatric disorders be conceptualized as "harmful dysfunctions" (J. C. Wakefield, 1992a, 1992b, 1999; S. O. Lilienfeld & L. Marino, 1995)? Wakefield's (1992a) central concept of disorder as "harmful dysfunction" is discussed by placing it in the context of a complementary discussion of disease, illness, the sick role, and evolution (D. F. Klein, 1978). S. O. Lilienfeld and L. Marino (1995) contended that proper biological function cannot be determined. This argument obscures the key significance of involuntary impairment of evolved functions. The claim that the Roschian concept has no counterpart in reality is incorrect and does not support the conclusion that dysfunctions are irreducibly evaluative and therefore arbitrary. J. C. Wakefield's (1999) views in this area are supplemented. The role of monothetic and polythetic categorization, extremal terms, and the concept of normality in nosology is considered. This analysis refutes the implication that deviance and illness are equivalent. The resolution of this debate is practically relevant to emphasizing areas of research investment, such as therapeutics. PMID- 10466265 TI - Harmful dysfunction and the DSM definition of mental disorder. AB - Physicians, including psychiatrists, give a lot of thought in their everyday work to answer the question of whether or not a particular patient has a disorder; they rarely give much thought to the broader issue of what constitutes a disorder. Remarkably, and consistent with the harmful dysfunction (HD) analysis, there is a broad consensus in both the general public and the medical and health professions as to what conditions are disorders--even though there is no consensus definition of disorder. The HD analysis is a substantial advance over previous attempts to define disorder in specifying the nature of what is not working in the individual (the dysfunction). The adoption of the HD analysis in DSM-V (Diagnostic and Statistical Manual of Mental Disorders, 5th ed.) would probably have little if any effect on the list of categories of mental disorders. Its main value would be in helping make revisions in the diagnostic criteria more valid as true indicators of disorder. PMID- 10466266 TI - Horsefeathers: a commentary on "evolutionary versus prototype analyses of the concept of disorder". AB - J. C. Wakefield's (1999) article further develops his harmful dysfunction (HD) model for disorder concepts. This commentary focuses on three areas. The first notes the imbalance in the debate between the Lilienfeld and Marino (1995) Roschian model and the HD model for disorder concepts. The second claims that Wakefield's purposes for the HD model have changed over the years and progressed toward irrelevance to psychopathology in general and toward irrelevance to actual nosologic, reimbursement, and sociopolitical controversies about disorder status. Further discussion is on how certain structural elements in Wakefield's arguments and current limitations of evolutionary theory permit a superficially attractive model for psychopathology. These arguments and limitations, however, harbor serious problems when confronted with actual disputes about disorders. The conclusion notes some virtues to Wakefield's inquiry, in style and substance. PMID- 10466267 TI - The abduction of disorder in psychiatry. AB - The evolutionary cornerstone of J. C. Wakefield's (1999) harmful dysfunction thesis is a faulty assumption of comparability between mental and biological processes that overlooks the unique plasticity and openness of the brain's functioning design. This omission leads Wakefield to an idealized concept of natural mental functions, illusory interpretations of mental disorders as harmful dysfunctions, and exaggerated claims for the validity of his explanatory and stipulative proposals. The authors argue that there are numerous ways in which evolutionarily intact mental and psychological processes, combined with striking discontinuities within and between evolutionary and contemporary social/cultural environments, may cause nondysfunction variants of many widely accepted major mental disorders. These examples undermine many of Wakefield's arguments for adopting a harmful dysfunction concept of mental disorder. PMID- 10466268 TI - Culture and context in the evolutionary concept of mental disorder. AB - The evolutionary theory of the concept of mental disorder as harmful dysfunction that J. C. Wakefield (1999) proposed (a) does not correspond to how the term disorder is used in psychiatric nosology or in clinicians' everyday practice; (b) does not cover the territory to which the term reasonably could be applied; and (c) is not especially useful for research, clinical, or social purposes. The broad concept of disorder is a polythetic, not a monothetic, concept. As such, there need be no essential characteristic, criterion, or single prototype of disorder. Instead, multiple prototypes with varying features are used to group together a wide range of disparate phenomena by analogy. Useful refinements of our concepts of disorder have come from analyses of the nature of action and intentionality. What are most needed now are careful analyses of the social embedding of our concepts in cultural knowledge and practice. PMID- 10466269 TI - Toward an evolutionary taxonomy of treatable conditions. AB - The definition of disorder as a harmful dysfunction (J. C. Wakefield, 1999) is a useful concept, anchored in the recognition that the evolved human architecture consists of a collection of functional mechanisms that may potentially be impaired and whose impairment may be harmful. Because natural selection organized each mechanism to solve a distinct adaptive problem under ancestral conditions, the criteria for whether a mechanism is dysfunctional are supplied by whether the mechanism has become impaired in performing its ancestral function. Because evolutionary function and dysfunction diverge markedly from normal human standards of value, many dysfunctions are beneficial, whereas various mechanisms that are performing their evolved function may cause disturbing outcomes. For this reason, many conditions in addition to disorders may require treatment, and the authors attempt to sketch an evolutionary taxonomy of treatable conditions. PMID- 10466270 TI - Mental disorder as a black box essentialist concept. AB - This is a reply to commentaries on the target article (J. C. Wakefield, 1999) on the evolutionary foundations of the concept of mental disorder in defense of the harmful dysfunction analysis (HDA) of disorder. The author argues that the HDA is adequate to explain disorder and nondisorder judgments and is not disconfirmed by any of the claimed counterexamples put forward by the commentators; the commentators' proposed alternatives to the HDA are inadequate to explain disorder and nondisorder judgments; and the concept of natural function is a factual, scientific concept, contrary to K. W. M. Fulford's (1999) claim that it is inherently evaluative. The foundations of the HDA are clarified by providing a black box essentialist analysis (H. Putnam, 1975; J. C. Wakefield, 1997, in press) of the concept of natural function that underlies the concept of disorder. PMID- 10466271 TI - Exaggerated anxiety is not unique to CO2 in panic disorder: a comparison of hypercapnic and hypoxic challenges. AB - Current biological models of panic disorder (PD) assert that this disorder is maintained by hypersensitivity to carbon dioxide (CO2) and related asphyxia cues, which is manifested as an exaggerated suffocation alarm (D. Klein, 1993). Because suffocation can result from both increased CO2 (hypercapnia) and decreased oxygen (O2; hypoxia), this study examined respiratory responding and anxiety during 5% CO2 (a hypercapnic challenge) and 12% O2 (a hypoxic challenge) in 14 PD patients and 14 matched controls (MC). Results demonstrate that irrespective of the source of suffocation, PD patients respond with increased anxiety relative to MC participants. Significant group differences were observed in respiratory functioning, with the PD patients showing higher respiratory rates in response to both challenges. The MC sample demonstrated the expected respiratory responses to both hypercapnia and hypoxia. The findings indicate that PD patients are hypersensitive to alterations in breathing and that this reactivity is not specific to CO2. PMID- 10466272 TI - First onset versus recurrence of depression: differential processes of psychosocial risk. AB - Differential risk factors for the onset of depression were prospectively examined in a community-based sample of adolescents (N = 1,709), some of whom had a history of major depressive disorder (MDD; n = 286) and some of whom did not (n = 1,423). From the theories of J. Teasdale (1983, 1988) and R. Post (1992) concerning the etiology of initial versus recurrent episodes of depression, the authors hypothesized that (a) dysphoric mood and dysfunctional thinking styles would be correlated more highly among those with a previous history of MDD than among those without a history of MDD; (b) dysphoric mood or symptoms and dysfunctional thinking would be a stronger predictor of onset of recurrent episodes (n = 43) than of first onsets (n = 70); and (c) major life stress would be a stronger predictor of first onsets of MDD than of recurrent episodes. The results provide support for the 3 hypotheses and suggest that distinct processes are involved in the onset of first and recurrent episodes of MDD. PMID- 10466273 TI - A longitudinal investigation of social causation and social selection processes involved in the association between socioeconomic status and psychiatric disorders. AB - Social causation theory and social selection theory have been put forth to explain the finding that low socioeconomic status (SES) is associated with risk for psychiatric disorders. The predictions of both theories were investigated using data from a community-based longitudinal study. Psychosocial interviews were administered to 736 families from 2 counties in New York State in 1975, 1983, 1985-1986, and 1991-1993. Results indicated that (a) low family SES was associated with risk for offspring anxiety, depressive, disruptive, and personality disorders after offspring IQ and parental psychopathology were controlled, and (b) offspring disruptive and substance use disorders were associated with risk for poor educational attainment after offspring IQ and parental psychopathology were controlled. These findings indicate that social causation and social selection processes vary in importance among different categories of psychiatric disorders. PMID- 10466274 TI - Major depression in the transition to adulthood: risks and impairments. AB - An ongoing longitudinal community study (N = 375) examined childhood risks and later adult impairments associated with 1-year Diagnostic and Statistical Manual of Mental Disorders (3rd ed., rev.; American Psychiatric Association, 1987) diagnoses of major depression during the transition to adulthood. Risks from birth to age 9 were reported by mothers, participants, and teachers. Teacher reported hostility at age 6 predicted later depression. At age 9, self perceptions of anxiety/depression, unpopularity, familial rejection, and abuse were potent risks. For men, neonatal and childhood health problems predicted later depression. For women, risks included family constellation, parental death, and poor academic achievement at age 9. Men and women who were depressed at age 18, age 21, or both demonstrated extensive psychosocial impairments in early adulthood, including poor overall functioning, interpersonal and behavioral problems, low self-esteem, and suicidality. PMID- 10466275 TI - Autobiographical memory processes and the course of depression. AB - The authors report a 6-month follow-up study of clinically depressed patients. At baseline, 2 indexes of autobiographical memory functioning were assessed: the presence of spontaneous intrusive memories of stressful life events and performance on the Autobiographical Memory Test (J. M. G. Williams & K. Broadbent, 1986), which measures overgeneral memory. The index of overgeneral memory was associated with greater levels of spontaneous intrusion of stressful memories. Overgeneral memory did not predict outcome, but depression at follow-up was predicted by the amount of intrusion and avoidance of stressful memories, even after controlling for initial severity of psychiatric symptoms. PMID- 10466276 TI - Implicit activation of alcohol concepts by negative affective cues distinguishes between problem drinkers with high and low psychiatric distress. AB - This study tested the hypothesis that negative affective (NEG) cues activate alcohol concepts more strongly in problem drinkers with high rather than low psychiatric distress (PD) and that a parallel difference applies to activation of NEG concepts by alcohol cues. A lexical decision task assessed memory activation in 36 problem drinkers with high vs. low scores on the Symptom Checklist-90. High PD drinkers displayed activation in each condition; low PD drinkers displayed inhibition. Activation of alcohol concepts by NEG cues correlated with intensity of PD, bias to drink in NEG states, and confidence to avoid drinking in these states. Results corroborate in vivo cue reactivity studies and indicate a functional difference between problem drinkers with high and low PD. PMID- 10466277 TI - Prospective evaluation of anxiety sensitivity in the pathogenesis of panic: replication and extension. AB - Increasing evidence suggests that anxiety sensitivity (AS) may be a premorbid risk factor for the development of anxiety pathology. The principal aim of this study was to replicate and extend a previous longitudinal study evaluating whether AS acts as a vulnerability factor in the pathogenesis of panic (N. Schmidt, D. Lerew, & R. Jackson, 1997). A large nonclinical sample of young adults (N = 1,296) was prospectively followed over a 5-week, highly stressful period of time (i.e., military basic training). Consistent with the authors' initial study, AS predicted the development of spontaneous panic attacks after controlling for a history of panic attacks and trait anxiety, and AS was found to possess symptom specificity with respect to anxiety versus depression symptoms. AS 1st-order factors differentially predicted panic attacks, with the Mental Concerns factor being the best predictor of panic in this sample. PMID- 10466278 TI - Testing Damasio's somatic marker hypothesis with psychopathic individuals: risk takers or risk averse? AB - Damasio and colleagues (A. R. Damasio, 1994; A. R. Damasio, D. Tranel, & H. Damasio, 1990) have theorized about a possible relationship between somatic markers and the behavior of psychopathic individuals (Ps), but, to date, there are no published data regarding the proposed relationship. The authors assessed 86 Caucasian and 71 African American male offenders using R. D. Hare's (1991) Psychopathy Checklist--Revised and used a modified version of Bechara and colleagues' (A. Bechara, A. R. Damasio, H. Damasio, & S. W. Anderson, 1994; A. Bechara, H. Damasio, D. Tranel, & A. R. Damasio, 1997) gambling task to test the hypothesis that Ps would, consistent with the somatic marker hypothesis, fail to become risk averse. Results indicated that level of anxiety, but not psychopathy, was predictive of response choices. Several limitations and implications of the study are noted. PMID- 10466279 TI - A state-of-the-profession report on midwifery in the managed care market. PMID- 10466280 TI - Midwifery in the 21st century. Recommendations from the Pew Health Professions Commission/UCSF Center for the Health Professions 1998 Taskforce on Midwifery. AB - Unprecedented changes in the delivery and financing of health care have produced angst and opportunity, criticism, and innovation. To explore the effects of these market-driven changes on midwifery, the University of California at San Francisco Center for the Health Professions convened a Taskforce on Midwifery in 1998. Consisting of eight experts from across the country, the Taskforce was charged with exploring the impact of health care system developments on midwifery, and identifying issues facing the profession and the roles midwives play in women's health care. The Taskforce answered its charge by offering 14 recommendations related to midwifery practice, regulation, education, research, and policy. The recommendations incorporate the Taskforce vision that the midwifery model of care should be embraced by, and incorporated into, the health care system in order to make it available to all women and their families. Midwives, educators, collaborators, and policymakers can use the recommendations to develop curricula, practice sites, and laws for an improved health care system that fully includes midwives and encompasses the midwifery model of care. PMID- 10466281 TI - Recent changes in birth attendant, place of birth, and the use of obstetric interventions, United States, 1989-1997. AB - Although more than nine out of every ten births are attended by physicians, the percent of births attended by midwives increased during the 1989 to 1997 period and accounted for 7% of all births in 1997. About 99% of births in 1997 were in hospitals, basically unchanged from 1989, but the percent of out-of-hospital births that occurred in residences increased over the period, while those in freestanding birthing centers declined. The percent of mothers receiving electronic fetal monitoring, ultrasound, and induction and stimulation of labor increased. The most dramatic increase was a doubling of the use of induction. In 1997, approximately 18% of all births were induced. Midwives as well as physicians increased the use of these obstetric procedures over the 1989 to 1997 period, and the use of many procedures by CNMs was as high, or nearly as high, as use by physicians. The rate of cesarean births dropped by 9%, from about 23% in 1989 to about 21% in 1997, while the rate of vaginal birth after a previous cesarean increased by 50%. The proportion of births assisted by forceps consistently declined during the period, while the use of vacuum extraction consistently increased. The number of episiotomies performed in the United States declined each year during the 1990-1996 period. PMID- 10466282 TI - Evidence-based practice and its application to childbirth care for low-risk women. AB - "Evidence-based medicine" has been hailed as the "new paradigm" for health care. This paper defines evidence-based practice, describes its development and growing importance, explains why randomized controlled trials are the "gold standard" for evidence about the effectiveness of specific therapeutic methods, warns about reaching conclusions based on any single study, and points the reader towards good sources of information on how to assess the relevance of findings from published studies and systematic reviews of the most reliable evidence regarding particular components of the care of pregnant women. As a result of those reviews, specific elements of perinatal care have been classified into categories based on their usefulness or harmfulness when applied to low-risk women. The paper goes on to summarize the evidence regarding three intrapartum practices that are demonstrably safe and useful and "should be encouraged" and four intrapartum practices that have trade-offs between beneficial and adverse effects and are "frequently used inappropriately." Some of the most beneficial elements of intrapartum care are not available to most women who give birth in American hospitals, and some practices that are useful but have adverse effects are being provided to an ever-expanding proportion of women. PMID- 10466283 TI - The midwifery model of care. AB - The midwifery and medical models for the care of pregnant women are based on particular perspectives on pregnancy and birth. The approaches resulting from these perspectives are complementary and, as a result of midwives and physicians working together, there has been significant merging of the models. Instead of two mutually exclusive ways of managing birth, there is wide variation. Nevertheless, there are important differences between the two models, including differences in philosophy and focus, in the relationship between the care provider and the pregnant woman, in the main focus of prenatal care, in use of obstetric interventions and other aspects of care during labor, and in the goals and objectives of care. The midwifery model has advantages for many women because it avoids unnecessary interventions during labor, thus helping the process remain normal, and because it addresses needs that are often not adequately met by the medical management model. PMID- 10466284 TI - Preserving midwifery practice in a managed care environment. AB - Since 1925 when nurse-midwifery emerged as a new health profession in the United States, and over the past 30 years under the auspices of the American College of Nurse-Midwives (ACNM), national standards for education, certification, and practice have enabled nurse-midwives to provide the quality of care that is highly valued by policy-makers and consumers. This paper describes the education and practice of over 7,000 midwives who have received national certification through mechanisms developed by the ACNM, describes the strengths of the profession, and reviews the impact of managed care on the practice of midwifery. Also highlighted is ACNM's development of a partnership, with the Maternal and Child Health Bureau, to study the impact of changes in the health care environment on pregnant women and ACNM certified midwives. This partnership is presented as an example of how the ACNM has evolved into an organization that is well-positioned to preserve midwifery in a managed care environment. PMID- 10466285 TI - Evolution and current status of direct-entry midwifery education, regulation, and practice in the United States, with examples from Washington State. AB - This paper describes the re-emergence of direct-entry midwifery in the United States, and focuses specifically on the over 1,000 midwives nationwide who are licensed in the 16 states where direct-entry midwifery is legal and regulated, and/or certified by the North American Registry of Midwives; it does not focus on direct-entry midwives or nurse-midwives who are certified by the American College of Nurse-Midwives Certification Council, Inc. Professional developments of direct entry midwives are highlighted, including the establishment of core competencies and articulation of values, the creation of a certification process, and development of education program accreditation. The current status of licensed midwives in Washington State, where state policies have supported the development of direct-entry midwifery and the integration of direct-entry midwives into managed care systems, is presented as one example of the evolution of professional direct-entry midwifery in this country. Additionally, recommendations from the UCSF Center for the Health Professions Taskforce on Midwifery, which address particular areas of concern for direct-entry midwives, are discussed. PMID- 10466286 TI - No free lunch on labor day. The risks and benefits of epidural analgesia during labor. AB - Epidural analgesia provides effective pain relief for women during labor. However, like all medical interventions, it also has potential side effects such as longer labor and a higher rate of intrapartum fever and operative vaginal delivery. A recent meta-analysis of randomized studies by Halpern et al concluded there was no association between epidural use and cesarean delivery. A critique of that meta-analysis, included in this paper, concludes that there are currently insufficient data to determine whether epidural analgesia leads to increased rates of cesarean delivery. This paper also presents results from several recent studies related to epidural analgesia conducted at Brigham and Women's Hospital in Boston. One study demonstrates a significant influence of prenatal planning on use of epidural during labor. Additional studies examine the strong association of epidural analgesia with intrapartum fever and the consequences of that fever for mother and infant. Epidural analgesia should remain an option available to women during labor. A more complete understanding of the risks and benefits that accompany its use is essential so that women and their care providers can make informed choices about pain relief during labor. PMID- 10466287 TI - What accounts for physician opinions about midwifery in Canada? AB - The objective of this study was to identify the characteristics of physicians practicing obstetrics in the province of Quebec, Canada and relate these to their opinions about midwifery practice. A self-administered questionnaire was sent to a systematic random sample of 844 physicians; 597 physicians answered (response rate = 71%). Results show that physicians who were more open to midwives had a more client-centered approach to maternity care. They had attended premed school outside Canada but received their specialty training in Canada. They also had often collaborated with midwives since they had begun practicing. Physicians who were more open to midwives were less demanding in terms of level of midwife training. They agreed that midwives should be self-regulating. It is concluded that greater knowledge of midwives' practice, gained through collaboration in the workplace and interdisciplinary education, could help physicians to better understand the significant contributions that midwives can make to the health care system. PMID- 10466288 TI - Cesarean section (VBAC) not safe. PMID- 10466290 TI - A model to assess age-related changes in two-point discrimination of plantar skin. AB - A device was constructed to assess two-point discrimination in the plantar skin of younger, middle-aged, and older adults. The device consisted of paired filaments, each delivering a tip force of 4.56 log units (equivalent to 3.63 g), at a fixed distance apart. Statistical analysis showed significant differences in two-point discrimination accuracy between these groups of subjects. Younger subjects showed greater accuracy of plantar two-point discrimination than either middle-aged or older subjects, with an apparent inverse linear relationship between subject age and plantar two-point discrimination ability. A model to assess age-related changes in two-point discrimination is presented. PMID- 10466289 TI - Mammalian birth. PMID- 10466291 TI - Hydroxyapatite as a bone substitute. AB - A bone substitute eliminates the need for autogenous and allogeneic bone grafting, along with the complications unique to each. Coralline hydroxyapatite is a synthetic bone void filler manufactured from marine coral, which has a natural trabecular structure similar to that of cancellous bone. While initial studies have been promising, the use of coralline hydroxyapatite may be limited in the foot and ankle owing to its inherent mechanical weakness and lack of biodegradation. In this retrospective study, 20 patients who received coralline hydroxyapatite implants were reviewed to determine associated complications and host reaction to the material. The results indicate that the material is biologically inert and safe to use as a small-defect filler in low-load applications. Long-term clinical trials are needed to determine the indications for this material in the foot and ankle. PMID- 10466292 TI - Clarifying proximal diaphyseal fifth metatarsal fractures. The acute fracture versus the stress fracture. AB - This article discusses the classification and treatment of proximal diaphyseal fifth metatarsal fractures. There are two types of proximal diaphyseal fracture of the fifth metatarsal: the acute proximal diaphyseal fracture and the proximal diaphyseal stress fracture. Confusion between the two types of fractures is probably due to their similar location and the historical practice of referring to all fractures in this location as Jones fractures. Both fractures are prone to delayed union and require long periods of nonweightbearing immobilization or internal fixation for healing. However, because the mechanism of injury is different for each fracture, the treatment plan may need to be tailored to the particular type of fracture. PMID- 10466293 TI - The effect of pregnancy on footprint parameters. A prospective investigation. AB - Pregnancy produces significant alterations in the posture of the pregnant woman; however, gait changes that occur during pregnancy have not been adequately evaluated in the literature. This prospective investigation of the footprints of 25 pregnant women from early pregnancy to just prior to parturition revealed a significant increase in the base of gait during walking. This change in gait function may be a compensatory mechanism to improve locomotor stability, and may have important implications for foot function and development of lower-extremity pathology in pregnant women. PMID- 10466294 TI - Phenol and alcohol chemical matrixectomy in diabetic versus nondiabetic patients. A retrospective study. AB - The authors performed a retrospective study of the safety and efficacy of toenail phenol and alcohol chemical matrixectomy in diabetic versus nondiabetic patients. Participating in the study were 66 patients, who underwent 137 nail procedures. The diabetic group consisted of 18 patients who underwent 39 procedures; the nondiabetic group consisted of 48 patients who underwent 98 procedures. The mean (+/- SD) healing time was 44 +/- 25 days for the diabetic group and 43 +/- 24 days for the nondiabetic group. The infection rate was 10.3% (4 of 39 procedures) for the diabetic group and 12.2% (12 of 98 procedures) for the nondiabetic group. In addition to validating this procedure for diabetic patients, this study showed no significant differences in the rate of postoperative complications between the two groups. PMID- 10466295 TI - Atypical heel pain. Hyperparathyroidism-induced stress fracture of the calcaneus. AB - The authors present a case of atypical heel pain masquerading as plantar fasciitis. The patient was subsequently diagnosed with hyperparathyroidism induced stress fracture of the calcaneus. The clinical entity of hyperparathyroidism and its manifestations in the skeletal system are presented, as well as a review of stress fractures and pertinent imaging studies. PMID- 10466296 TI - Dermatofibrosarcoma protuberans. A rare case involving the pediatric foot. AB - Dermatofibrosarcoma protuberans is a rare, slow-growing, locally destructive, intermediate-grade malignancy that originates in the dermal layer of the skin. Dermatofibrosarcoma protuberans rarely metastasizes and has a marked tendency to recur following local surgical excision. The lesion is found most commonly on the trunk, on the proximal aspect of both the lower and upper extremities, and in the head and neck region. Dermatofibrosarcoma protuberans occurs most commonly in people between the ages of 20 and 50. This article describes the clinical and pathologic features of this rare, malignant neoplasm and describes a rare case of dermatofibrosarcoma protuberans involving the pediatric foot. PMID- 10466297 TI - Onychocryptosis associated with the treatment of onychomycosis. AB - With the advent of effective oral therapies for onychomycosis, there have been anecdotal reports of an increased incidence of onychocryptosis associated with the use of these new agents. The authors undertook a chart review of 100 consecutive patients treated for onychomycosis in a private medical practice to determine the incidence of onychocryptosis. A total of 37 of these patients (37%) developed paronychia ranging from simple pain to a severe inflammatory response with redness, drainage, and granuloma formation. Of these, 19 patients (19%) required surgical procedures to control onychocryptotic symptoms. Podiatric physicians should be aware that ingrown toenails may be an adverse consequence of effective treatment for onychomycosis. PMID- 10466298 TI - Integrating strategies for podiatric public health, clinical services, and education. AB - Changes in the health-care system will continue to modify the delivery of podiatric medical care and podiatric medical education. Podiatric medicine and its educational programs will need to focus on the management of care, special populations, and disease prevention. Future providers will need to participate as team members and respond to new patterns of care, including the special needs of the older population. This article explores methodologies for professional activities, administration, incentives for change, academic issues, innovation, implementation, outcome measures, and health policy development. Podiatric medicine must be prepared to establish standards, protocols, and guidelines to serve as the benchmark for foot health. PMID- 10466299 TI - Clinical diagnosis of posterior tibial tendon rupture. PMID- 10466300 TI - Superglue for the treatment of heel fissures. PMID- 10466301 TI - Subungual squamous cell carcinoma mistaken for a verruca. PMID- 10466302 TI - [Role of nuclear medicine in organ transplantation]. AB - It has been widely accepted that the organ transplantation has become one of the choices for the therapy of end-stage diseases (kidney, heart, liver, lung, etc.). However, organ transplantation requires extremely high cost and heavy psychological burden for his/her family. Therefore, and moreover to reward the donor's good will, it must be highly successful. Nuclear medicine tests can offer reproducible functional information with and without images. Repeat tests can be performed easily. These characteristics are suitable not only for preoperative evaluation of donors and recipients but also for monitoring the transplanted organs. Therefore, nuclear medicine can potentially play an important role in organ transplantation. Competitive diagnostic modalities such as US, CT, MR are also progressing and very useful in some situation in organ transplantation. Nuclear medicine physicians and technologists should be aware of the merits and the role of nuclear medicine in organ transplantation and should try to prepare for good service to provide useful information. This article reviews the role of nuclear medicine in organ transplantation, demonstrating representative cases. PMID- 10466303 TI - [The calculated reference value of the tubular extraction rate in infants and children: an attempt to use a new regression equation]. AB - This study was designed to investigate the empirical tubular extraction rate (TER) of the normal renal function in childhood and then propose a new equation to obtain TER theoretically. The empirical TER was calculated using Russell's method for determination of single-sample plasma clearance and 99mTc-MAG3 in 40 patients with renal disease younger than 10 years of age who were classified as having normal renal function using diagnostic criteria defined by the Paediatric Task Group of EANM. First, we investigated the relationships of the empirical value of absolute TER to age, body weight, body surface area (BSA) and distribution volume. Next we investigated the relationships of the empirical value of BSA corrected TER to age, body weight, BSA and distribution volume. Linear relationship was indicated between the absolute TER and each body dimensional factors, especially regarding to BSA, its correlation coefficient was 0.90 (p value). The BSA-corrected TER showed a logarithmic relationship with BSA, but linear regression did not show any significant correlation. Therefore, it was thought that the normal value of TER could be calculated theoretically using the body surface area, and here we proposed the following linear regression equation: Theoretical TER (ml/min/1.73 m2) = (-39.8 + 257.2 x BSA)/BSA/1.73 The theoretical TER could be one of the reference values of the renal function in the period of the renal maturation. PMID- 10466304 TI - [Chronological study for solitary bone metastasis in the sternum from breast cancer with bone scintigraphy]. AB - Since breast cancer is frequently associated with bone metastasis, bone scintigraphies have been performed to determine pre-operative staging and to survey postoperative bone metastasis. The sternum, in particular, is a site at which is difficult to differentiate between benign bone disease and bone metastasis, because of varied uptake and wide individual variations. In this study, chronological bone images were scintigraphied in six cases with solitary sternal metastasis and three cases with benign bone disease including two fracture cases and one arthritis case. On bone scintigrams in which solitary sternal metastasis appeared, increased uptake was found in five cases, and photon deficiency was observed in one case. During follow-up scintigraphies, abnormal accumulations, such as hot spots and cold lesions, increased in the bone metastasis while abnormal uptake disappeared or was unchanged in the benign bone disease cases. On CT, four cases showed osteolytic change, and one exhibited osteosclerotic change. These findings indicate that sternal metastasis usually shows osteolytic change, even if a hot lesion is recognized on bone scintigraphy. In solitary sternal metastasis, for which early diagnosis is difficult, both an integrated diagnosis using other imaging techniques and chronological bone scintigraphy are important. PMID- 10466305 TI - [Evaluation of parathyroid imaging methods with 99mTc-MIBI--the comparison of planar images obtained using a pinhole collimator and a parallel-hole collimator]. AB - Parathyroid scintigraphy with 99mTc-MIBI was performed using two kinds of collimators, namely, a pinhole one and a parallel-hole one, to evaluate which one was more suitable for the detection of hyperfunctioning parathyroid lesions. In the studies using 99mTc source, the pinhole collimator showed better efficiency and spatial resolution in the distance where the parathyroid scan are actually performed. In the phantom study, the nodular activities modeling parathyroid lesions were visualized better on the images obtained using the pinhole collimator. In clinical studies for 30 patients suspicious of hyperparathyroidism, hyperfunctioning parathyroid nodules were better detected when the pinhole collimator was used. In conclusion, the pinhole collimator was thought to be more suitable for parathyroid scintigraphy with 99mTc-MIBI than the parallel-hole collimator. PMID- 10466306 TI - [Assessment of left ventricular contraction abnormalities with myocardial infarction using gated technetium-99m sestamibi SPECT: comparison of wall thickening and regional ejection fraction analysis for the detection of coronary artery stenosis]. AB - ECG-gated myocardial Technetium-99m sestamibi SPECT is a useful technique to measure myocardial perfusion and function simultaneously. In this study, wall thickening (WT) and regional ejection fraction (rEF) using ECG-gated SPECT have been studied to determine which parameter would be more sensitive to detect coronary artery stenosis in patients with acute myocardial infarction (AMI). Forty-five patients (36 men, 9 women, mean age 63 +/- 9 years old) with AMI were examined. CAG was performed for all patients. ECG-gated SPECT was performed 60 min after the intravenous injection of 555 MBq 99mTc-sestamibi at rest. Commercially available software (QGS) was used to produce WT and rEF polar maps from acquired SPECT data. The WT and rEF polar maps were evaluated visually and quantitatively. WT indicated higher sensitivity (80.3% vs. 59.1%, p < 0.05) and accuracy (86.7% vs. 74.8%, p < 0.05) than rEF for detecting overall coronary artery stenosis on visual interpretation. On quantitative analysis, WT had higher specificity (91.3% vs. 75.4%, p < 0.05) and accuracy (85.9% vs. 72.6%, p < 0.05) than rEF for detecting overall coronary artery stenosis, and showed a higher specificity (93.8% vs. 59.4%, p < 0.01) and accuracy (88.9% vs. 62.2%, p < 0.01) for detecting LCX stenosis. Moreover, sensitivity of WT for detecting coronary artery stenosis without infarction was higher than that of rEF significantly in quantitative analysis (75.0% vs. 31.3%, p < 0.05). These results suggested that WT was superior to rEF for detecting the coronary artery stenosis in patients with and without myocardial infarction. We concluded that WT is more sensitive indicator to determine localization of regional left ventricular dysfunction in AMI than rEF. PMID- 10466307 TI - [Assessment of regional myocardial function by ECG-gated myocardial SPECT]. AB - PURPOSES: Regional myocardial functional parameters were assessed by ECG-gated myocardial SPECT analysis in normal subjects and ischemic heart diseases. METHODS: Normal subjects (13 male and 10 female) and 51 patients with chronic ischemic heart disease underwent ECG-gated myocardial SPECT. A dose of 740 MBq of 99mTc-MIBI was injected at rest, and gated SPECT was performed 60 min later. Wall motion (WM) and systolic wall thickening (WT), % tracer uptake were evaluated by quantitative gated SPECT program (QGS). Regional parameters were obtained in the 16 segments based on the functional polar map. In the normal group, standard values were evaluated in the lateral, septal, anterior and inferior regions both in male and female subjects. In the ischemic heart disease group, sensitivity and specificity of these parameters were assessed in each segment. To estimate the ability of WM and WT in detecting regional dysfunction, decreased perfusion area, which was defined as < mean - 2SD by the normal profile, was used as a standard. The receiver operating characteristics (ROC) area analysis was also performed. RESULT: In the normal profile, % tracer uptake was decreased in the anterior segments of female group, no significant difference was observed between male and female in WM and WT. WM was decreased in the septum and increased in the lateral segment. WT didn't show any difference regional difference. Sensitivity and specificity of WM were 56%/91% in the anterior, 0%/100% in the septum, 43%/87% in the inferior, 31%/85% in the lateral. WT were 67%/93% in the septum, 67%/79% in the inferior, and 59%/81% in the septum. The area under ROC curve was WM 0.63, WT 0.85 (p < 0.005 between WM and WT) in the septum, in inferior WM 0.77, WT 0.80 (p = 0.57), in anterior WM 0.86, WT 0.87 (p = 0.095), in lateral WM 0.68, WT 0.78 (p = 0.037). CONCLUSION: In normal profile, the % tracer uptake in the anterior wall decreased in females, but WM and WT did not show significant difference in each region affected this influence. The septal WM was decreased in the normal profile and ability to diagnose regional function was also decreased compared with WT. Thus, we can conclude that WT is preferable for detecting septal functional abnormality. PMID- 10466308 TI - [Usefulness of 67Ga scintigraphy in deciding surgical indication in secondary hyperparathyroidism]. AB - In order to evaluate the usefulness in deciding surgical indication in secondary hyperparathyroidism (SHP), 67Ga scintigraphy was performed in 37 patients of SHP before parathyroidectomy (PTx). The radionuclide accumulation in skull and submandible was classified into 4 patterns (skull-submandibular pattern, skull pattern, submandibular pattern and normal pattern). Serum alkaline phosphatase levels were significantly elevated in patients of skull-submandibular pattern (13 cases) compared with skull pattern (6 cases), submandibular pattern (6 cases) and normal pattern (12 cases). Serum intact parathyroid hormone levels were significantly elevated in patients of skull-submandibular and skull patterns compared with normal pattern. No significant difference was observed among the weight of resected parathyroid glands. In 4 of 6 patients of normal pattern on 67Ga scintigram, bone scintigraphy showed a characteristic pattern of SHP including an increased accumulation in the skull and submandible. Bone mineral density (BMD) in the distal radius was increased within six to twelve months after PTx in 10 of 11 patients of skull-submandibular pattern on 67Ga scintigram, whereas only one patient showed an increase in BMD in 9 patients of normal pattern. In summary, it was concluded that 67Ga scintigraphy could provide a useful information in deciding the indication for PTx in secondary hyperparathyroidism. PMID- 10466309 TI - [Reduction of the influence of the liver uptake to the myocardial uptake on technetium-99m myocardial SPECT; usefulness and problems of a mask processing method]. AB - The aim of this study is to evaluate the usefulness of a mask processing method for obtaining the true myocardial tracer distribution by eliminating the influence of the liver uptake to the myocardial uptake on myocardial SPECT images by using technetium-99m (99mTc) blood flow agents. A SPECT imaging was performed with a two-head SPECT system (GCA-7200A/DI) in both phantom and clinical studies. The mask processing method was applied to the reconstructed and projection images. The phantom consisted of heart, lung, liver and spine. A defect was located in the inferior wall of the left ventricle and other parts of the heart and liver were filled with 99mTc solution. For clinical study 10 patients with difficulty in the interpretation of the inferior wall were selected for the evaluation of usefulness of the mask method. In the phantom study, the mask processing method applied to the reconstructed images was able to remove the overlapped liver from the heart, but was not able to remove the influence of the liver uptake to the myocardial uptake. Nevertheless, the mask processing method applied to the projection images successfully eliminated not only the overlapped liver but also the influence of the liver uptake to the myocardial uptake. In the clinical study, the liver uptake could be removed from the uptake in the inferior wall in 8 of 10 patients with the mask processing methods. In 2 patients, the overlapped liver uptake could not be eliminated from the uptake in the inferior wall because the distance between the liver and heart was too short. The mask processing method applied to the projection images was thought to be superior to that applied to the reconstruction images in both phantom and clinical studies. The mask processing method, especially applied to the projection images, seems to be useful for the elimination of the liver uptake from the inferior wall of the myocardium on myocardial SPECT images using 99mTc blood flow agents. PMID- 10466310 TI - [Development of 18F-FDG ([F-18]-2-fluoro-2-deoxy-D-glucose) injection for imaging of tumor reflecting glucose metabolism--results of preclinical studies]. AB - Fluorine-18-2-fluoro-2-deoxy-D-glucose (18F-FDG) injection was prepared by a modification of a method originally developed by Hamacher et al. The dosage form is the injectable solution (2 ml) containing 185 MBq of 18F-FDG at a calibration time. Preclinical studies of the agent were performed. Its radiochemical purity is more than 95% and expiration time is 4 hours after the calibration time at ambient temperature. No toxicity was observed with up to 200 mg/kg and 100 mg/kg of non-radioactive FDG intravenously injected to rats and dogs in single-dose toxicity tests, respectively. Biodistribution studies demonstrated that the radioactivity was mainly distributed into brain (3.0 to 3.3% I.D./Organ at 30 minutes) and heart (4.2 to 5.8% I.D./Organ at 1 to 3 hours) after intravenous injection of the agent to normal rats. In a tumor transplanted mouse model (colon 26), tumor uptake was 10.9 +/- 3.5% I.D./g at 1 hr after intravenous injection of the agent, the radioactivity was retained until 3 hours. The radiation absorbed dose was estimated according to the MIRD Pamphlet based on the biodistribution data both in humans reported by Mejia et al. and rats described in this report. The radiation absorbed dose was not higher than those of commercially available radiopharmaceuticals. In conclusion, the 18F-FDG injection is expected to be useful for further clinical application. PMID- 10466311 TI - The forensic detective. PMID- 10466312 TI - The case of R. v. Sheila Bowler. PMID- 10466313 TI - Changes to Section 2 of Sexual Offences Act 1976. AB - Rape complaints are at their highest since statistics began, whilst the rape conviction rate is at an all-time low. The frequency with which the complainant's previous sexual history is admitted at trial is perceived as a major factor. The government intends to legislate to reduce this, in accordance with either the Scottish or the New South Wales' legislative models. This article analyses the case law, argues that neither proposed model will suffice and advocates that the Canadian legislation should, with adaptations, be enacted in England. PMID- 10466314 TI - 'A scandalous and corrupt hospital'--reflections on the Fallon report. PMID- 10466315 TI - Section 25 aftercare under supervision: the first eighteen months' experience. AB - Supervised discharge was introduced, against much professional opposition, in the Mental Health (Patients in the Community) Act 1995. All patients subject to supervised discharge (SD) in a large health district were identified by medical records departments, and the outcome of their SD on 1 October 1997, eighteen months after the legislation came into effect, was assessed. In that first eighteen months, half of the general, rehabilitation and forensic psychiatrists in the trust used SD for 22 patients. Of the 12 orders in effect for over six months, six were renewed at six months and three at one year. In spite of professional hostility and scepticism about the clinical practicalities of SD, the use of the new power has gradually increased with experience. Outcomes to date seem generally positive, but further definitive research is needed. PMID- 10466316 TI - Psychiatric disorders in first-degree relatives of patients with opiate dependence. AB - High rates of psychiatric disorders, particularly major depression, alcoholism, drug-use disorder and antisocial personality disorder in the relatives of opiate dependent patients compared with normal controls, have been reported in some previous studies. This study was designed to evaluate the prevalence rates of drug-use and other psychiatric disorders in the first-degree relatives of opiate dependent patients and compare these with rates in the first-degree relatives of surgical and psychiatric patients. A case-control study was conducted to compare the prevalence of psychiatric and drug-use disorders (which were estimated by the Family History Method) in 241 biological first-degree relatives of 50 opiate dependent patients to that in 235 and 222 first-degree relatives of 50 surgical and 50 psychiatric patients respectively. The main outcome measure was the relative risk (expressed as odds ratio) of psychiatric and drug-use disorders. First-degree relatives of opiate-dependent patients were found to have significantly higher rates of drug-use and antisocial personality disorders, compared with relatives of surgical and psychiatric probands. The odds ratio for alcoholism appeared significantly higher only in first-degree female relatives of opiate users, suggesting effect modification by sex. The study provides further evidence for the higher rate of drug-use disorder in the first-degree relatives of opiate-dependent patients. These findings suggest that familial drug-use disorder contributes to a vulnerability to opiate misuse. Implications of these findings for the classification and treatment of opiate dependence are discussed. PMID- 10466317 TI - Prevalence of cannabinoids in urine samples from forensic autopsies. AB - Urine samples from 393 forensic autopsies were tested for the presence of cannabinoids by means of the radio-immunoassay test kit from DPC, Los Angeles. Fifteen per cent were found positive, mainly men and young adults. Use of cannabis was found to be closely related to drug addiction, and thus in cannabis positive drug addicts, non-natural manners and causes of death dominate. In non addicts the prevalence of cannabis use was 7.5%, and in non-addicted cannabis users, manners and causes of death were not very different from those found in cannabis-negatives. PMID- 10466318 TI - A study of methadone in fatalities in the Strathclyde Region, 1991-1996. AB - There was a substantial increase in the percent of drug screens testing positive for methadone between 1991 and 1996 in the Strathclyde region of Scotland. Seventy-nine per cent (n = 136) of these deaths were drug-related, involving methadone either alone or in combination with other drugs such as diazepam, temazepam, alcohol and morphine. The involvement of methadone in the majority of these fatalities was due to diversion of legitimate supply. This paper highlights the dangers of resuming methadone consumption following a period of abstinence or when taken in combination with other drugs. PMID- 10466319 TI - Carbon monoxide fatalities in medicolegal autopsies. AB - All post-mortem medicolegal reports issued by Jordan University Hospital (JUH) during the period 1978-96 were reviewed. Carbon monoxide (CO) as a silent killer was responsible for 31.5% (n = 110) of poisoning fatalities. These were due to unintentional exposure to CO sources in confined spaces (bedrooms (24.6%), worker lodgings (23.6%) and bathrooms (16.4%), and without the victims being aware of CO hazards. Sixty-five per cent of CO fatalities occurred during December to February and 30% of the cases involved the age group 20-29 years, followed by the age group 30-39 years (20%). Non-Jordanians constituted 50% of CO fatalities and Egyptian workers accounted for 78%. Being unaccustomed to cold climates, and on low incomes, they had been using simple and unsafe heating devices. Many of these deaths could have been prevented. Safety educational campaigns to increase awareness of the general population of CO sources, their hazards and how to avoid them should be held continuously during the cold months of the year. All health and safety sectors are advised to be involved. PMID- 10466320 TI - The contribution of forensic pathology to clinical cardiology. PMID- 10466321 TI - Suicide among psychiatric in-patients with schizophrenia in an Australian mental hospital. AB - Psychiatric in-patient suicides are associated with schizophrenia. In this paper, 62 suicides in patients with schizophrenia, over a 21-year period in a large psychiatric hospital in Melbourne, Australia, were examined. The characteristics, including demographic and clinical data, for the suicides were compared with a comparison group of 22 'alive' in-patients with schizophrenia. Suicide among in patients with schizophrenia was associated with previous deliberate self-harm, a greater number of episodes of previous deliberate self-harm, pre-admission and intra-admission suicidal thoughts, intra-admission suicidal attempts, fluctuating suicidal ideation, longer length of stay, a greater number of ward transfers, and prescription of a greater number of neuroleptics and antidepressants. Over 40% of suicides occurred after absconding from hospital and a similar number during periods of approved leave. Violent methods (including jumping in front of trains, trams and road traffic, jumping off buildings, hanging and drowning) were most frequently used. It is concluded that psychiatric units should be developed away from readily available methods of suicide. In-patients at high risk should be observed carefully to avoid absconding and suicide. Suicide risk should be examined carefully in patients prior to approving leave, particularly if they have fluctuating suicidal ideation. PMID- 10466322 TI - The moon and mental health. PMID- 10466323 TI - Third time lucky--an unusual suicide. AB - We present the case of a 39-year-old male who died with three significant and separate shotgun wounds. During the investigation, the possibility of murder was considered, but reconstruction of the case and post-mortem findings led to a coronial conclusion that the death was a suicide, accounted for by the type of weapon used and the stamina of the deceased. PMID- 10466324 TI - Homicide by paraquat poisoning. AB - We report on two cases of homicide by paraquat poisoning. Both cases occurred in children and the common clinical presentations were gastrointestinal ulceration and acute respiratory distress, with pneumomediastinitis. Bullous emphysema was a common autopsy finding. Physicians and pathologists should be aware of these findings, for they are almost diagnostic of acute paraquat poisoning. PMID- 10466325 TI - The management of opiate addicts in police custody. PMID- 10466326 TI - [Regulation of excitotoxicity and apoptosis as future treatment modality for various ocular diseases]. PMID- 10466327 TI - [Rapid pulse injection of indocyanine green angiography for evaluation of choroidal venous circulation]. AB - PURPOSE: To investigate the influence of the injection duration on the early phase of indocyanine green fundus angiography, we performed indocyanine green fundus angiography using a rapid pulse injection method, and compared early-phase findings with those obtained using a previous injection method. METHODS: Rabbit eyes were used. Normal eyes (10), eyes with chorioretinal atrophy (5), and eyes with occlusion of vortex veins (5) were examined. RESULTS: In normal eyes, rapid pulse injection resulted in visualization of some small bright fluorescent boluses from 6 to 12 seconds after injection, whereas with the previous method they were unclear. In eyes with occlusion of vortex veins, rapid pulse injection resulted in visualization of delayed hyperfluorescence of choroidal veins against the background, and the region of delayed venous filling could easily be determined. In eyes with chorioretinal atrophy, only the rapid pulse injection clearly revealed delayed filling of choroidal veins. CONCLUSIONS: The rapid pulse injection method enabled detection of lesions which were not clearly visible using the previous injection method, and is useful for study of choroidal circulation. PMID- 10466328 TI - [The uptake of fluoroquinolone agent drugs by cultured human conjunctival epithelial cells and cultured SIRC rabbit corneal cells]. AB - PURPOSE: To investigate the uptake of drugs by cultured human conjunctival epithelial cells (HCEC) and cultured rabbit corneal cell lines (SIRC). METHOD: The drugs examined were ofloxacin (OFLX), lomefloxacin (LFLX), and norfloxacin (NFLX). The amount of drug uptake in the cultured cells that were exposed to the drugs was measured by high-performance liquid chromatography (HPLC). RESULT: The amount of LFLX, OFLX, and NFLX uptake in HCEC was 1.38 +/- 0.21 nmol/10(6) (mean +/- standard deviation) cells (5 min), 0.79 +/- 0.05 nmol/10(6) cells (5 min), and 0.56 +/- 0.03 nmol/10(6) cells (5 min), respectively. The amount of LFLX, OFLX, and NFLX uptake in SIRC was 1.08 +/- 0.18 nmol/10(6) cells (5 min), 0.58 +/ 0.17 nmol/10(6) cells (5 min), and 0.41 +/- 0.35 nmol/10(6) cells (5 min), respectively. The uptake levels of LFLX by both cultured cells were higher than those of OFLX and NFLX. CONCLUSION: This method may be useful as a screening method for intraocular drug dynamics studies. PMID- 10466329 TI - [Effect of vitamin A palmitate on the synthesis of mucins in cultured conjunctiva]. AB - PURPOSE: To evaluate the effect of vitamin A palmitate on the synthesis of mucins from goblet cells in cultured rabbit conjunctiva. METHODS: Palpebral conjunctiva was obtained from 12 white rabbits and was cultured in serum-free RCGM 2 which was developed for culture of rabbit corneal epithelial cells. This method eliminated retinol in the serum. High-performance liquid chromatography was used in quantitating N-acetylneuraminic acid contained in the sugar chain as an indicator of mucin content. RESULTS: Addition of vitamin A palmitate to the cultured medium resulted in significant increase in the amount of N acetylneuraminic acid in the medium. CONCLUSIONS: The increase in N acetylneuraminic acid in the cultured medium suggests that the increase of goblet cells in cultured conjunctiva is related to the synthesis of mucins due to vitamin A. RCGM 2 as a culture medium promises to be useful in evaluating the pharmacological activity of vitamin A. High-performance liquid chromatography facilitated quantitation of mucins in the culture medium. PMID- 10466330 TI - [Effects of a new excitotoxic amino acid, dysiherbaine, on cultured Muller cells]. AB - PURPOSE: To determine pharmacological response of dysiherbaine on cultured Muller cells considering the glutamate receptor functions. Dysiherbaine is a new excitotoxic amino acid, which was recently isolated from the water extract of a certain sponge. SUBJECTS AND METHOD: Retinas of adult rabbits were used to prepare the Muller cells. Intracellular calcium ion concentration ([Ca2+]i) analysis was done by fluorophotometry with calcium indicator, Fura-2 AM. RESULTS: A transient increase of [Ca2+]i was observed following the administration of dysiherbaine (2.5 microM -2.5 mM), but it was not observed in the extracellular calcium-free solution. This increase was blocked by the non NMDA glutamate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). A dysiherbaine induced increase in [Ca2+]i following preincubation of the NMDA glutamate receptor antagonist, (5 R, 10 S)-(+)-5-methyl-10, 11-dihydro-5 H-dibenzo [a, d] cyclohepten-5, 10-imine hydrogen maleate (MK 801) was seen in the same number of Muller cells as without the antagonist. CONCLUSION: Dysiherbaine appears to act primarily as a non NMDA glutamate receptor agonist having a secondary action as a NMDA glutamate receptor agonist. PMID- 10466331 TI - [Blood flow velocity in the central retinal artery of exotoxin-induced uveitis in rabbits]. AB - PURPOSE: Exotoxin-induced uveitis in rabbit eyes was established by intravitreal inoculation of staphylococcal enterotoxin B (SEB). Association between findings of pulsed Doppler assessments and histological features were evaluated. MATERIAL AND METHODS: Eyes of rabbits were unilaterally inoculated with 50 ng/kg of intravitreal SEB. Pupil diameter, blood flow velocity in the central retinal artery in both eyes and heart rate were measured using an ultrasonic diagnostic apparatus after inoculation. RESULTS: The pupil diameter and the mean blood flow velocity in central retinal arteries decreased and reached a trough 3-5 hours after the inoculation, then increased. Pulsatility index showed reciprocal fluctuations. Histological features at the early phase after inoculation revealed lymphocytic, plasmacytic, and polynuclear leukocytic infiltrations around the anterior chamber, limbal conjunctivae, ciliary body, and nerve fiber layer of the retina, followed by recovery after about 5 days. At the late phase, recurrent inflammations in the ciliary body and intravitreal space were observed in addition to invasion into the rod and cone layer. CONCLUSION: Early and late phase uveitis was established by intravitreal SEB. It is considered that the uveitis may be caused by two different regulatory mechanisms. PMID- 10466332 TI - [Three cases of carotid cavernous sinus fistulas where the main ocular manifestation was restricted ocular motility]. AB - BACKGROUND: Some cases of carotid cavernous sinus fistulas (CCFs) have few ocular congestive signs and symptoms. These cases need to be accurately diagnosed as having CCFs. CASES: Three patients with CCFs without ocular congestive signs or symptoms developed unilateral abducens restriction and visited our clinic. FINDINGS: Color Doppler imaging revealed retrograde flow of the superior ophthalmic veins bilaterally in the three cases. The diagnose of CCF was confirmed by cerebral angiography. CONCLUSION: It is sometimes difficult to diagnose CCF, when a patient without ocular congestive signs or symptoms shows disturbed ocular motility as the only ocular manifestation. In such cases, color Doppler imaging may show the presence of CCF. PMID- 10466333 TI - [Acquired progressive esotropia and acquired strabismus fixus]. AB - BACKGROUND AND PURPOSE: We previously reported on acquired convergent strabismus fixus and its incomplete type of esodeviation, which we named acquired progressive esotropia. In this study, we tried to confirm that the esotropia was acquired and progressive and to determine the most appropriate surgical procedure. SUBJECTS AND METHODS: Forty-three cases, 32 of acquired progressive esotropia and 11 of acquired convergent strabismus fixus with severe myopia, were rechecked to evaluate their clinical and physiological features. Thirty-seven cases underwent strabismus surgery and the surgical results were also evaluated. RESULTS: We obtained positive proof that the esodeviation was an acquired situation and progressed into convergent strabismus fixus from some pictures at a young age. The condition of some of these patients developed into convergent strabismus fixus in a short time. Combined recession-resection operation and transposition of superior and inferior rectus muscles were effective for the patients without severe abducting disorders. CONCLUSION: We must make a certain diagnosis of progressive esotropia in the early stages, because the condition of some of these patients will develop into strabismus in a short time if we miss the appropriate time for surgery. PMID- 10466334 TI - [Clinical results of selective laser trabeculoplasty]. AB - PURPOSE: Selective laser trabeculoplasty (SLT) is a new laser procedure using a frequency-doubled Q-switched Nd: YAG laser (wavelength: 532 mm). The laser parameters are set to selectively target pigmented trabecular meshwork (TM) cells without coagulative damage to the TM structure or non-pigmented cells. We investigated the safety and efficacy of SLT in lowering intraocular pressure (IOP). SUBJECTS AND METHOD: Sixty-seven eyes of 67 patients with uncontrolled open angle glaucoma were treated with the Coherent Selecta 7000 (Coherent Inc., Palo Alto, CA). Nineteen of 67 patients had previously received argon laser trabeculoplasty (ALT). A total of approximately 60 non-overlapping spots were placed over 180 degrees of the TM ranging from 0.5 to 1.0 mJ per pulse. The maximum energy level at which no bubble formation was observed determined choice. RESULTS: The average preoperative IOP was 22.4 mmHg. Six months after the operation, mean IOP reduction was 4.4 mmHg, and mean outflow pressure (OP) reduction was 38.1%. One month after the operation 68.7% of patients responded to treatment with an OP reduction of at least 20% ("responders"). Transient IOP elevation of 5 mmHg or greater was seen in 25.4% of patients. The success rate at 6 months after operation was 64.6% for all patients (67 eyes) and 78.2% for the responders (46 eyes). An analysis using a Cox proportional hazard model showed that a low preoperative IOP was the significant determinant for success, and the hazard ratio for the IOP increase of 5 mmHg was 2.12. Other factors such as age, gender, past history of ALT, and goniopigment were not significantly related to success. CONCLUSION: SLT appears to be a safe and effective way to lower IOP. PMID- 10466335 TI - [Two cases of frosted retinal angiitis with central retinal vein occlusion]. AB - BACKGROUND: Frosted retinal angiitis usually occurs in children, and has a good prognosis. We report two cases of unilateral frosted retinal angiitis in adults. They resulted in visual degradation because of associated central retinal vein occlusion and neovascular glaucoma. CASES: Case 1 was a 36-year-old female. Almost all retinal veins and some retinal arteries had vasculitis in her right eye, and the veins were slightly dilated and sheathed. Case 2 was a 23-year-old female. Angle hypopyon was observed in her left eye. Retinal veins were dilated, meandered, and sheathed. Retinal hemorrhages were also observed. In both cases, systemic steroid therapy gradually improved the retinal vasculitis, but central retinal vein occlusions gradually developed, and in spite of systemic administration of urokinase and panretinal photocoagulation, neovascular glaucoma developed, and visual acuity became degraded in both cases. CONCLUSION: Two cases of frosted retinal angiitis complicated by retinal vein occlusion were reported. Careful observation of retinal blood flow is necessary in frosted retinal angiitis in adults. PMID- 10466336 TI - [Kallmann syndrome: a failure of GnRH neuronal migration]. PMID- 10466337 TI - [Clinical study of Helicobacter pylori infection]. PMID- 10466338 TI - [Effects of bright light on cognitive disturbances in Alzheimer-type dementia]. AB - We investigated the effectiveness of bright light therapy on cognitive disturbances and its effect on circadian (sleep-wake) rhythm in Alzheimer-type dementia (ATD). Twenty-seven patients with ATD were treated with bright light therapy in the morning for 4 consecutive weeks. We evaluated the cognitive functions and circadian rhythms of the patients as a whale, and as members of two groups (one: questionable and mild dementia: the other: moderate and severe dementia; both groups classified by the severity criteria of Clinical Dementia Rating). We assessed circadian rhythms by actigraphy and cognitive states by Mini Mental-State Examination (MMSE) and Alzheimer's Disease Assessment Scale (ADAS) before and after light therapy. Bright light therapy improved circadian rhythm. Although bright light therapy had no Significant effect on the Severity of dementia, it improved the MMSE scores, cognitive functions of ADAS scores (memory > language) and non-cognitive functions of ADAS scores (behavior = mood), especially in the questionable and mild dementia group. These results suggest that bright light therapy improves cognitive functions with the modification of circadian rhythm, especially in the early stages of ATD. PMID- 10466339 TI - [Autoimmune fatigue syndrome and fibromyalgia syndrome]. AB - We have encounted two patients with fibromyalgia (FM) initially diagnosed as having autoimmune fatigue syndrome (AIFS). To investigate the relationship between AIFS and FM, the distribution of the tender points in patients with AIFS was assessed according to the ACR criteria for FM. It was revealed that AIFS patients had 5.6 tender points on averages. Patients with headaches, digestive problems, or difficulty going to school had more tender points than patients without. Patients with ANA titers < 1: 160 had more tender points than patients with ANA > or = 1: 160. Anti-Sa negative patients had more tender points than positive patients. These results suggest a relationship between AIFS and FM in terms of the pathophysiologic mechanisms of the numerous tender points. In other words, ANA-positive FM patients could be one form of AIFS, as well as ANA positive chronic fatigue syndrome patients. Thus, autoimmunity could explain the controversial disease entities of FM and/or CFS. PMID- 10466340 TI - Value of magnetization transfer contrast as a sensitive technique to reflect histopathological changes in the white matter adjacent to the frontal horns of lateral ventricles. AB - The purpose of this study is to evaluate the usefulness of magnetization transfer contrast (MTC) as a technique to reflect histopathological changes in the white matter adjacent to the frontal horns of the lateral ventricles. Radiological pathological correlation was performed in six patients who underwent Magnetic Resonance (MR) examination prior to death and in whom postmortem examinations of the brain were obtained. The extent and the severity of degeneration in the white matter adjacent to the frontal horns were evaluated histopathologically, and compared with those observed on the conventional proton density (PD) weighted MR images (Group 1). Changes in the white matter of another 35 patients were classified into three types according to the pattern of high signals adjacent to the frontal horns on conventional PD weighted MR images, and magnetization transfer ratio (MTR) in the white matter adjacent to the frontal horns was calculated from multi-slice and single-slice FSE images (Group 2). The relationship between signal intensities and MTR in the white matter adjacent to the frontal horns was evaluated. The extent of degeneration in the white matter adjacent to the frontal horns was classified into mild, moderate and severe types on the basis of stainin for myelins, axons and astrocytes. In Group 1, histopathological findings indicated a difference in severity of degeneration in the white matter adjacent to the frontal horns among the three types, while no significant differences were noted in the signals on PD weighted MR images. In Group 2, MTR showed significant differences in the signal intensities in the white matter adjacent to the frontal horns (p < 0.01) between the three types, while conventional PD weighted MR images failed to differentiate between them. In conclusion, MT imaging is a sensitive technique to evaluate the histopathological changes in the white matter adjacent to the frontal horns that cannot be detected by conventional MR imaging. PMID- 10466341 TI - Two types of peritoneal dissemination of pancreatic cancer cells in a hamster model. AB - Peritoneal dissemination has an unfavorable impact on the prognosis of pancreatic cancer, and a peritoneal dissemination model was created in hamsters by using an experimental pancreatic cancer to clarify its pathological characteristics. PGHAM 1, a cancer cell line we established from BOP induced pancreatic cancer in Syrian golden hamsters, was inoculated into the abdominal cavity of Syrian golden hamsters. After inoculation, sequential changes in the diaphragm, omentum, and parietal peritoneum, and the metastatic patterns of the PGHAM-1 cells were morphologically investigated by macroscopical, microscopical, and ultrastructural observation. The cancer cells were easily absorbed at the stomata in the diaphragm and milky spots in the omentum, which were absorptive lymphatic structures, and lymphatic metastasis occurred 4 days after inoculation. In the parietal peritoneum, however, the cancer cells attached to and proliferated on the parietal peritoneum where mesothelial cells had exfoliated and the basement membrane was exposed. This process was comparatively time-consuming, and metastasis occurred in the parietal peritoneum at 7 days after inoculation. This study suggested that there might be two patterns of peritoneal dissemination of hamster pancreatic cancer. One route is lymphatic metastasis via stomata in the diaphragm and milky spots in the omentum, and the other is direct metastasis on the parietal peritoneum; each metastasis forms independently. PMID- 10466342 TI - Repeated propofol anesthesia for a patient with a history of neuroleptic malignant syndrome. AB - Neuroleptic malignant syndrome (NMS) is the most serious side effect produced by the administration of antipsychotic drugs. NMS shares many clinical similarities with malignant hyperthermia (MH), but the etiology of NMS and the relation between NMS and MH remain unknown. Anesthetic regimens for patients with NMS are not well established. We gave repeated anesthesia to a patient with a history of NMS undergoing electroconvulsive therapy for the treatment of depression. Propofol and vecuronium were used in twelve consecutive ECT sessions without complications. In this case report, we describe the safe and satisfactory repeated use of propofol in a patient with a history of NMS, and outline NMS and its questionable relation to MH. PMID- 10466343 TI - Prenatal diagnosis of acute massive fetomaternal hemorrhage. AB - We present here 2 cases of acute and 2 cases of chronic massive fetomaternal hemorrhage. A sinusoidal fetal heart rate pattern may indicate chronic fetomaternal hemorrhage, but, when increased variability is observed in fetal monitoring, maternal hemoglobin F should be measured to exclude acute fetomaternal hemorrhage. PMID- 10466344 TI - [Research on the influences that affect the physician's perception of discharge summaries. A survey of outpatient department physicians]. AB - The object of this study was to improve the completion of discharge summaries. The purpose of this article is: 1. to describe the perceptions of outpatient department physicians as to whether incomplete discharge summaries affect outpatient medical care or not, and 2. to elucidate the factors contributing to their perceptions. The physicians were randomly selected from among "the National Certified Training Hospitals" and "Japan Society for Quality in Health Care", and the size of the hospitals, the medical record management system and the format of the discharge summary were chosen for analysis of the factors that affect physicians' perceptions. Of the 2,304 questionnaires sent, 935 (40.6%) were returned. The results showed that 832 (89.0%) of the physicians thought that outpatient medical care would be affected if discharge summaries were not completed. Based on the analysis of the factors affecting the perceptions of the physicians, there was a trend showing that the influence was lower in small hospitals (p < 0.05), that the centralized management system for medical records had a higher influence (p < 0.05), and that the format of the discharge summary also affects physician's perceptions (p < 0.05). PMID- 10466345 TI - [Diagnosis and treatment of atopic dermatitis]. PMID- 10466346 TI - [Geriatric care and economic evaluation]. PMID- 10466347 TI - [Recent progress in angiogenesis research]. PMID- 10466348 TI - [Osteoporosis]. PMID- 10466349 TI - [Pathology and treatment of anemia in the elderly]. AB - An important background characteristic of anemia in the elderly is decrease in hematopoiesis due to aging. Factors influencing hematopoiesis in the elderly include changes in the distribution of hematopoietic tissue, changes in hematopoietic stem cell density and changes in the hematopoietic inductive microenvironment. In the present study, in order to assess changes in the bone marrow with aging, the fat tissue area, uncleated cell-count and cellularity in the bone marrow, in addition to changes in the diameter of the vascular lumen which result primarily from sclerotic changes in the dorsomedial artery of the bone marrow were determined in different age groups. The results revealed that all of the aforementioned factors changed significantly with aging. We also describe on the results of assays of inflammatory cytokines (IL-1, IL-6, TNF alpha), lactoferrin and transferrin receptors in cases of anemia of chronic disorders (ACD) which own secondary to chronic inflammatory diseases and is known to frequently afflict the elderly. PMID- 10466350 TI - [A pathomorphological study on atherosclerotic regression with references between apoptosis and atherosclerotic plaque with central depression in rabbit aorta]. AB - Atherosclerotic plaque with central depression (depressed lesions) have been proposed as one morphological feature of atherosclerotic regression in the elderly. We have also revealed that depressed lesions have been found not only in the aorta of elderly human but also in rabbit aorta. In this study a relationship between apoptosis and atherosclerotic regression was immunohistochemically studied to clarify the possibility of the pathogenesis of depressed lesion in rabbit aorta. Twenty male New Zealand White rabbit (NZW) were fed with a 1.0% cholesterol diet during a 12-week atherogenic induction period. Then they were fed only with a basic diet for 36-week (n = 6) and 48-week (n = 6, experimental group) regression periods. A control group was fed with 1.0% cholesterol diet for 12-week (n = 2) and 60-week (n = 6). They were killed and aortas were fixed with formalin. Sections obtained from aortas were processed for histological and immunohistochemical studies including the TUNEL method, staining with Ki-67 and others. Several depressed lesions were found in the aortas of NZW animal, but none were noted in control aortas. Results of surface involvement in the experimental groups were significantly lower than in the control, and the aortas of the experimental group had atherosclerotic regression. Apoptosis was found in the depressed lesions, which had more apoptotic cells than non-regressed atherosclerotic plaques. Furthermore, the apoptotic cells were significantly greater in the center of depressed lesions than in marginal areas. Ki-67 staining was positive in the atherosclerotic plaque, but negative in the depressed lesions. It appears that the NZW aortic atherosclerotic plaques may transform to depressed lesions with apoptosis. Atherosclerotic regression has been associated with apoptosis. PMID- 10466351 TI - [Age-related change in the alpha-tocopherolquinone/alpha-tocopherol ratio in the rat erythrocyte membrane]. AB - alpha-Tocopherol (alpha-Toc), a lipophilic phenolic antioxidant that is localized mainly in the biomembrane, protects cells against oxidation-associated cytotoxicity by prevention of membrane lipid peroxidation, maintenance of the redox balance intracellular thiols and stabilization of the membrane structure. We investigated the age-related changes in redox dynamics of alpha-Toc in plasma and erythrocyte membrane of an elderly (66 weeks old) and young group (10 weeks old). Total, alpha-, beta + gamma-, delta-Toc and alpha-tocopherolquinone (alpha TocQ) in plasma and erythrocyte membrane were determined by high-performance liquid chromatography (HPLC) with a series of multiple coulometric working electrodes (CWE). Rat venous blood sample was divided into plasma and erythrocyte layers by centrifugation, and then erythrocyte membrane sample was prepared according to the method of Dodge et al. under a stream of nitrogen. In plasma, total and alpha-Toc concentrations were increased, and beta + gamma-, delta-Toc and alpha-TocQ concentrations were decreased age-dependently. In the erythrocyte membrane, total, alpha-TocQ concentrations and three fractions of tocopherols decreased age-dependently. Also, a decrease in the alpha-TocQ/alpha-Toc ratio in erythrocyte membrane was observed in the elderly group. These findings suggest that the alpha-Toc uptake in erythrocyte membrane and utilization rate of alpha Toc in erythrocyte membrane decline age-dependently. This decline may promote membrane lipid peroxidation. alpha-Toc redox dynamics in erythrocyte membrane were useful to investigate the pathophysiology of aging mechanisms related to oxidative stress. PMID- 10466352 TI - [Multicenter prospective survey of prognosis of hypertensive elderly outpatient under antihypertensive treatment--morbidity and mortality of cardiovascular diseases and cancer]. AB - Although calcium-channel blockers and angiotensin-I-converting enzyme inhibitors are often used for treatment of hypertension in the elderly in Japan compared to those in the United States and in European countries, there have been few investigations on the prognosis of the elderly receiving these antihypertensive treatments. The Research Group for "Guidelines on the Treatment of Hypertension in the Elderly" collaborated with the Comprehensive Research Projects on Aging and Health group of the Ministry of Health and Welfare of Japan in performing a 3 year survey on the outcome of 700 hypertensive elderly outpatients (> or = 60 years) receiving treatment of antihypertensive drugs. Antihypertensive drugs including dihydropyridine-type calcium channel blockers, beta blockers, angiotensin-I-converting enzyme inhibitors, diltiazem, diuretics and old-type antihypertensives (hydralazine, budralazine, and centrally acting drugs such as clonidine, methyldopa and guanabenz) were administered to 71.3%, 30.4%, 26.2%, 14.0%, 8.6%, and 6.4% of the 642 elderly patients surveyed for three years, at the time of registration, respectively. Morbidity and mortality rates of total cerebro-cardiovascular diseases, stroke, and heart diseases, were 27.6 and 7.81/1,000 patient-years, 15.1 and 3.6/1,000 patient-years, 10.4 and 4.2/1,000 patient-years, respectively. These results were similar or even better than those of megatrials of antihypertensive treatments for elderly patients in Europe and United States. After adjustment for potential confounding factors, multiple logistic analysis revealed that a past history of ischemic heart disease, use of the old-type antihypertensive drugs, male gender, and diastolic high blood pressure were independent risk factors for the morbidity of cerebro cardiovascular diseases taking the group of non-cerebro-cardiovascular disease as the reference group. We also identified 22 cases of newly occurred malignancies including 7 fatal cases. However, none of the antihypertensives was significantly related to the occurrence of malignancies. These results lead support to the tendencies in the use of antihypertensive drugs in Japan. PMID- 10466353 TI - [Association of silent cerebral infarction with cerebral hemispheric--sub tentorial infarction in patients with supra-nuclear dysarthria or dysphagia]. AB - Little information is available concerning the latent effects of silent cerebral infarction (SCI) on dysarthria (DA), dysphagia (DP), or both. A detailed analysis of MR images of multiple cerebral infarction was carried out to determine whether SCI is associated with the development of DA or DP. In this study MR images of the supra- and sub-tentorial regions were obtained from 14 patients presenting with persistent DA and DP (DA + DP group) and 9 patients presenting with DA alone (DA group) after the first episode of cerebral infarction. The DA + DP inducing lesion was identified from the change in signal intensity and the side with symptoms in 6 patients. Involvement of 3 lesions of the bilateral cortical branches, striatum, and pons on the line connecting the contra-lateral SCI with the lesion were noted in 4 patients (67%, vs. 40% for the DA group). Latent association of SCI with the development of supra-nuclear DA and DP was noted in 1 patient each from the DA + DP and DA groups. The results of this study support the concept that SCI patients include those in which SCI is involved in the development of supra-nuclear DA or DP, and suggest that SCI should be treated. PMID- 10466354 TI - [A follow-up study on the outcome and relevant factors in senile dementia of Alzheimer type and vascular dementia]. AB - We followed 126 patients with senile dementia of Alzheimer type (SDAT), and 129 over age 65 with vascular dementia (VD) who were diagnosed at the Center for Elderly Dementia in our institution between February 1990 and February 1993. At 5 year follow-up, 62 patients with SDAT and 71 patients with VD had died. These patients were assessed prospectively to investigate the neuropsychiatric and somatic factors related to the prognosis of SDAT and VD. There were no significant differences in the average age at onset and time of diagnosis of dementia between the SDAT and VD groups. Mean age at death, mean duration of dementia and 75% survival duration from dementia onset were shorter in patients with VD than in those with SDAT. Pneumonia was the most common cause of death in patients with either SDAT or VD, followed by geromarasmus in the SDAT group, and cerebrovascular and cardiovascular diseases in the VD group. In patients with SDAT, a poor prognosis was correlated with severe dementia, impaired intellectual function and degree of cortical atrophy. In patients with VD, physical complications, impaired motor function and hypoproteinemia enhanced the probability of death. This study confirmed that a progressive neurodegenerative course and physical conditions such as motor dysfunction or malnutrition are closely associated with the prognosis of patients with SDAT and VD, respectively. PMID- 10466355 TI - [Three cases with low levels of serum copper due to long-term enteral nutrition]. AB - Three bed-ridden patients who had had only one kind of enteral nutrition without sufficient copper element during a few years showed very low levels of serum copper. Two of them also had leukopenia. The abnormal findings disappeared after the feeding nutrients rich in copper element. The leukopenia may have been due to copper deficiency rather than zinc deficiency. We confirmed that long-term parenteral nutrition must contain trace elements, for example copper. PMID- 10466356 TI - [An elderly case of giant infected liver cyst that decreased dramatically without percutaneous aspiration]. AB - A 70-year-old woman was referred to our hospital for treatment of cholelithiasis. A giant liver cyst (6 cm in diameter) had been diagnosed three years earlier. On admission, she had low grade fever and hepatomegaly. High values were observed for WBC (9900/microliter), CRP (8.9 mg/dl), GPT (45 IU/l), ALP (1399 IU/l), gamma GTP (333 IU/l) and LAP (249 IU/l). The diagnosis of infected liver cyst (8 cm in diameter) was made based on contrast-enhanced CT scan. Endoscopic retrograde cholangiopancreaticography showed no communication between the cyst and the intrahepatic bile duct. She was successfully managed with antibiotics and discharged without percutaneous aspiration the cyst. On abdominal CT scan 4 months after the discharge, the liver cyst had decreased dramatically in size (1 cm in diameter). The patient remains healthy without symptoms. PMID- 10466357 TI - [A case of myelodysplastic syndrome associated with IgA nephropathy]. AB - A 72-year-old man was admitted for examination of dyspnea and pitting edema of the lower legs in July, 1996. His hemoglobin level was 6.9 g/dl, and myelodysplastic syndrome (MDS) was revealed by bone marrow aspiration, and frequent transfusions were needed. His renal function rapidly deteriorated in the middle of August (BUN 45 mg/dl, Cr 4.8 mg/dl) and IgA nephropathy (IgAN) with marked intestinal nephritis was disclosed by renal biopsy. In November, joint manifestations of warmth and pain, which suggested arthritis, appeared at the bilateral wrist and ankle joints. Soon after receiving prednisolone (20 mg/day), the arthritis was relieved. Renal function also improved (BUN 41 mg/dl, Cr 21 mg/dl) and frequent transfusions were no longer necessary. This is a case with various clinical manifestations of MDS, IgAN, and arthritis, and appears to be the first MDS case complicated with IgAN. A number of case reports have identified immune abnormalities in patients with MDS. Immune and bone marrow abnormalities have been reported to be involved in the pathogenesis of IgAN. Thus, MDS could be complicated by IgAN. Their pathogenetic association is discussed in this paper. PMID- 10466359 TI - 27th Annual meeting of the American Society for Photobiology. Washington, DC, USA. July 10-15, 1999. Abstracts. PMID- 10466358 TI - [XVI National Congress of the Spanish Society of Parenteral and Enteral Nutrition. Las Palmas de Gran Canaria, 12-14 May 1999. Abstracts]. PMID- 10466360 TI - Estimation of loads in the lumbar spine during slow symmetrical lifts and a method for their reduction. PMID- 10466361 TI - Type of motion and lubricant in wear simulation of polyethylene acetabular cup. AB - The wear of ultra-high molecular weight polyethylene, the most commonly used bearing material in prosthetic joints, is often substantial, posing a significant clinical problem. For a long time, there has been a need for simple but still realistic wear test devices for prosthetic joint materials. The wear factors produced by earlier reciprocating and unidirectionally rotating wear test devices for polyethylene are typically two orders of magnitude too low, both in water and in serum lubrication. Wear is negligible even under multidirectional motion in water. A twelve-station, circularly translating pin-on-disc (CTPOD) device and a modification of the established biaxial rocking motion hip joint simulator were built. With these simple and inexpensive devices, and with the established three axis hip joint simulator, realistic wear simulation was achieved. This was due to serum lubrication and to the fact that the direction of sliding constantly changed relative to the polyethylene specimen. The type and magnitude of load was found to be less important. The CTPOD tests showed that the subsurface brittle region, which results from gamma irradiation sterilization of polyethylene in air, has poor wear resistance. Phospholipid and soy protein lubrication resulted in unrealistic wear. The introduction of devices like CTPOD may boost wear studies, rendering them feasible without heavy investment. PMID- 10466362 TI - Development of a ten-station, multi-axis hip joint simulator. AB - Joint simulators are now used extensively to evaluate the performance of materials and designs of total replacement hip and knee joints. In this Technical Note a new ten-station hip joint simulator with biaxial rotational articulation synchronized to a physiological loading cycle is described. The current simulator manufactured by ProSim Limited (Manchester, UK) is a development of a first generation machine designed and built in-house at DePuy International Limited (Leeds, UK). The use of this new form of ten-station hip simulator to evaluate the performance of 22 mm zirconia femoral heads and ultra-high molecular weight polyethylene acetabular cups over some 7 million loading cycles is described elsewhere [1]. PMID- 10466363 TI - An axisymmetric contact model of ultra high molecular weight polyethylene cups against metallic femoral heads for artificial hip joint replacements. AB - Contact mechanics of ultra high molecular weight polyethylene (UHMWPE) cups against metallic femoral heads for artificial hip joints is considered in this study. Both the experimental measurement of the contact area and the finite element prediction of the contact radius, maximum contact pressure and maximum Von Mises stress have been carried out for a wide range of contemporary artificial hip joints. Good agreement of the contact radius has been found between the experimental measurements and the finite element predictions based upon an elastic modulus of 1000 MPa and a Poisson's ratio of 0.4 for UHMWPE material under various loads up to 2.5 kN. It has been shown that the half contact angle for all the cup/head combinations considered in this study is between 40 degrees and 50 degrees under a load of 2.5 kN. The importance of this result has been discussed with respect to the anatomical position of the cup when placed in the body and the selection of a simple wear-screening test for artificial hip joints. The predicted contact radius and maximum contact pressure from the finite element model have also been compared with a simple elasticity analysis. It has been shown that the difference in the predicted contact radius between the two methods is reduced for more conforming contacts between the femoral head and the acetabular cup and smaller UHMWPE cup thickness. However, good agreement of the predicted maximum contact pressure has been found for all the combinations of the femoral head and the acetabular cup considered in this study. The importance of contact mechanics on the clinical performance of artificial hip joint replacements has also been discussed. PMID- 10466364 TI - The influence of continuous sliding and subsequent surface wear on the friction of articular cartilage. AB - Reciprocating motion friction tests were conducted upon cartilage-on-metal contacts while subjected to a constant load. Initial friction coefficients were compared with repeat friction coefficients following a sufficient load removal period. The repeat friction coefficients were marginally higher than the initial values and both were primarily dependent on the loading time. It was concluded that while a wear component had been identified, which modestly increased friction coefficients, the overriding parameter influencing friction was loading time. The authors postulate that fluid phase load carriage (being dependent on loading time) within the articular cartilage is largely responsible for low friction coefficients in the mixed and boundary lubrication regimes. This mechanism has been referred to as biphasic lubrication. Both synovial fluid and Ringer's solution were used as lubricants. Over the assessed 120 min loading time friction coefficients rose from 0.005 (for both lubricants) after 5 s to 0.50 and 0.57 for synovial fluid and Ringer's solution respectively. Synovial fluid was found to significantly reduce friction coefficients compared to Ringer's solution over broad ranges of the assessed loading times (p < 0.05). Stylus and non contacting laser profilometry were successfully employed to provide reliable, quantitative and accurate measures of surface roughness. Laser profilometry before and after a continuous sliding friction test revealed a significant increase in surface roughness from Ra = 0.8 (+/- 0.2) micron to Ra = 2.1 (+/- 0.2) microns, (p < 0.0005); confirming that surface wear was occurring. Scanning electron microscopy (SEM) revealed the typical highly orientated collagen fibres of the superficial tangential zone. Environmental SEM (ESEM) of fully hydrated cartilage specimens provided largely featureless images of the surface which suggested that sample preparation for conventional SEM was detrimental to the authenticity of the cartilage surface appearance using SEM. Two distinct acellular, non-collagenous surface layers were identified using ESEM and transmission electron microscopy (TEM); respectively referred to as the boundary layer and surface lamina. The phospholipid/glycoprotein based boundary layer will provide boundary lubrication during intimate contact of opposing cartilage surfaces. The surface lamina, being a continuum of the proteoglycan interfibrillar matrix, is present to prevent fibrillation of the underlying collagen fibres. Both layers may contribute to the time dependent frictional response of articular cartilage. Although laser profilometry did reveal surface wear which was consistent with a small increase in friction, the primary variable controlling the friction coefficient was the period of loading. PMID- 10466365 TI - Quantification of interdigitation at bone cement/cancellous bone interfaces in cemented femoral reconstructions. PMID- 10466366 TI - Feasibility study on a composite material construction technique for highly stressed components in reciprocal walking orthoses for paraplegic patients. AB - Reciprocal walking for thoracic level paraplegic patients using reciprocal walking orthoses has become a routine treatment option. Two general design options are currently deployed within an overall treatment regime. Research has shown that one has better walking efficiency but is cosmetically less acceptable to the patient. Design analysis and experimental data have shown that a major factor in improved walking efficiency is the lateral stiffness of the body brace section of the orthosis. This is the area where problems of cosmesis in the more efficient orthosis are perceived because of the employment of metallic structures. The use of composite material structures to achieve shapes which are more closely conforming to the patient is an attractive option. However the brittle nature of these materials makes it unlikely that the requirement for the ductile failure mode will be achieved from a straightforward moulding. A new construction technique has been devised which has the potential to provide a safe failure mode with greater stiffness and lighter weight. This feasibility study has been undertaken to demonstrate its potential so that further work can be justified which will provide sufficient evidence to support a patent application. The successful outcome of the study, in which stiffness was increased by 60 per cent with a weight reduction of 50 per cent and a failure mode comparable with the original metal structure, suggests that further work will enable the dilemma in the choice of orthosis to be resolved. PMID- 10466367 TI - An investigation of the performance of Biostop G and Hardinge bone plugs. AB - Although distal plugging is a common procedure to prevent distal flow of polymethyl-methacrylate (PMMA) cement during cementing of femoral prostheses, there is little biomechanical testing to confirm that (a) the plugs do not displace under cementing pressure, and (b) they do in fact occlude cement flow. Two designs of femoral intramedullary plugs, the Biostop G (Bioland, France) and Hardinge (De Puy, Leeds, UK) were examined to determine their performance under cement pressurization in a biomechanical test. A testing rig was fabricated in which distal migration could be measured as a function of cement pressurization. Sectioning of the samples after polymerization of the cement revealed the extent of cement flow. The results show that, even in this well controlled test, there is significant variability in plug performance. It is shown that the Biostop G displaces less than the Hardinge for similar cement pressures. Sectioning reveals that cement can escape around the Hardinge plug at high pressures. Furthermore, a pore forming effect of the Biostop G plug was occasionally observed indicating that design improvements may be possible for this plug. PMID- 10466368 TI - [The European Project on Improvement of Heart Failure. The Diagnostic Study Group of the Working Group on Heart Failure of the European Society of Cardiology]. PMID- 10466369 TI - [The organization and structure of pediatric cardiology. A bet for the future?]. PMID- 10466370 TI - Haemodynamics during liver transplantation in familial amyloidotic polyneuropathy: study of the intraoperative cardiocirculatory data of 50 patients. AB - BACKGROUND: Circulatory instability frequently complicates liver transplantation for familial amyloidotic polyneuropathy (FAP) and may be a source of surgical morbidity and mortality. OBJECTIVE: To evaluate FAP intraoperative haemodynamic data and their relation to the duration of surgery, and need for anaesthetic drugs. RBC and sympathomimetic amines. SETTING: Clinical study during a four year period. PATIENTS (mean +/- SD): Group I included 50 consecutive FAP ATTR Met 30 recipients of first transplantation. Age was 35.3 +/- 7.1 years, neurological score 34.3 +/- 13 in 100 and time elapsed from first symptom 5.0 +/- 2.7 years. Group II (control), not different concerning age and sex, included 51 patients transplanted during the same period with other pathologies. METHOD: Anaesthetic protocol, monitoring and surgical techniques were similar in both groups. Data of the two groups were compared either by the Student's t-test or Fisher's exact test. RESULTS: Low values of systemic vascular resistance index were observed in both groups, with no differences between them. Systemic arterial pressures were usually lower in group I, because cardiac index and heart rate were also significantly lower, although within normal values. However, in group I, isoflurane (a vasodilator anaesthetic) was used during less time (p < 0.05) and in lower concentrations (p < 0.01) and phenylephrine was necessary in 26% of patients vs 0 patients in group II (p < 0.001). CONCLUSION: FAP patients presented a different intraoperative behaviour when compared to other patients submitted to liver transplantation. From a clinical point of view, the authors stress: 1--As a result of autonomic dysfunction, the administration of anaesthetic drugs to FAP patients always presents the risk of producing significant hypotension; even the use of ketamine does not prevent hypotension; 2 -Safety is ensured by beat-to-beat surveillance of arterial pressures and the capacity to act immediately to support circulation; 3--These patients seem to be very sensitive to decreases in the pre-load; 4--Hypotension is also frequent with an adequate pre-load, usually as the result of low SVR; an infusion of a vasoconstrictor drug emerges as the most frequent treatment requested and our experience supports it as an effective one. PMID- 10466371 TI - [Atheroma plaque of the carotid bifurcation: how to identify an "active" lesion?]. AB - The identification of carotid atheromatous plaques associated to a higher neurologic risk may be important in therapeutic decision making for asymptomatic patients and symptomatic patients with 50%-70% stenosis. The introduction of high definition ultrasonography (HDU) and computer-assisted image analysis provides the possibility of a standard, objective and detailed characterization of the structure of the carotid plaque. The aim of this study is to analyse the relationship between the ultrasonographic characteristics of a group of plaques and the risk of associated cerebrovascular events and cerebral infarction. MATERIAL: One hundred carotid bifurcation plaques (in 68 patients) were studied for the presence of ipsilateral cerebrovascular events. In 61 patients (87 plaques), a correlation with CT scan for the presence of cerebral infarction was possible. METHOD: The lesions were studied by HDU (ATL-HDI 3000) and digitalized for computer-assisted standardization of the image. The analysis included the appreciation of the histogram of the image gray-scale pixel distribution by use of commercial software Adobe Photoshop 3.0. The parameters analysed for global echogenicity were the median of the histogram and the percentage of percentile 40 hypoechogenic pixels (40). Statistical analysis was made with STATA 4.0 software with categorical variables analysed by chi-square and Fisher's exact test and continuous variables analysed by variance analysis and Student's t test. RESULTS: Thirty eight (38%) plaques were symptomatic and 34 (39.1%) were associated to cerebral infarction. The degree of stenosis was > 70% in 51%; between 50 and 69% in 27% and < 50% in 22%. The mean of the median and P40 values was 33.9 and 60.3% in the symptomatic and 46.8 (p = 0.005) and 46.6% (p = 0.001) in the asymptomatic plaques respectively. In the plaques associated to cerebral infarction, it was 32.7 and 61.6% respectively, and in the ones with negative CT scan, it was 44.6 (p = 0.005) and 48.1% (p = 0.002). The mean of the median in the plaques vs. degree of stenosis was: > 70%--33.3; 50-69%--45.1; < 49%--57.7 (p < 0.001). In the series the cut-off point for the median value was 32 and for P40 it was 43 (for any degree of stenosis): G1--plaques < 32 (echolucent); G2--plaques > 32 (echogenic). In G1 symptoms occurred in 60% of the plaques and in 26% of the plaques in G2 (p = 0.0001). CT scan was positive in 66% of the echolucent plaques and in 25% of the echogenic plaques (p = 0.0238). CONCLUSION: 1. The more echolucent plaques are associated with a significantly higher neurological risk. 2. The plaques associated with higher degrees of stenosis are more echolucent. 3. The use of a standard and objective methodology in the analysis of the echographic structure of carotid plaques is important and limits the known intra and inter-observer variability of subjective appreciation. PMID- 10466372 TI - [The prevention of cardiovascular diseases]. PMID- 10466373 TI - [A coronary disease registry. The fundamental basis for its control]. AB - A register may be a list in which each item is individually identified, corresponding to uniform information about individual persons, collected in a systematic way, in order to serve predetermined purposes. However, disease registration, more than "notification", requires a permanent record for cases to be followed-up and statistical analysis to cover the frequency and survival. The register requires information to be up-to-date for individual and general disease surveillance. Nevertheless, on establishing a register, it is fundamental to define the purposes within the main areas of interest: identification of individuals; protection of the individual; surveillance; epidemiology (incidence; distribution, etiology, diagnosis, prognosis, prevention, analysis of benefits and risks); planning and evaluation of health care services; evaluation of treatment; research; education; development of methodology. The advantages of developing a coronary heart disease register are multiple: standardization of methods, accessibility to a database resulting from a multicentric study (inter center cooperation and participation), definition of the problem magnitude, acquisition of a large range of concrete knowledge. The aims of registers determine the data to be recorded and the analysis. Before starting a Myocardial Infarction Register, or a register for another specified disease, we need to establish some questions: Who should be registered? What data should be recorded? How many cases? Where can they be established?, and when? The functioning of a register is based on the following steps: definition of criteria introduced; mode of detection; initial examination; follow-up (annual?) and computer system. The organization and maintenance of a disease register is not easy (in terms of time, money and personnel). Therefore, a successful register should be established when the previous conditions are fulfilled and if the required information (aims) cannot be obtained by other means. Even so, problems may occur in case-finding (sub-registration is the principal risk) and certainty of diagnosis or during the follow-up (data loss and patient drop-out). It is essential to maintain the efforts to guarantee the quality of the data and the surveillance of the patients that entered the study. Thus, a coronary heart disease (myocardial infarction) register may be fundamental for its control and, simultaneously, it forms an important part of the health information systems. PMID- 10466374 TI - [The successes and failures in the primary prevention of coronary disease]. AB - The modification of major behavioural and environmental risk factors has helped reduce coronary heart disease. It seems, however, that prevention results were not so good as expected. It is assumed that the major cause of this apparent unsuccessful data is basically due to world population ageing, as it is known that coronary heart disease is much more frequent in older people. Therefore the current life expectancy shows that the preventive measures have indeed been successful. From the analysis and discussion of the preventive epidemiological recommendations and the identification of new risk factors, the Authors suggest new strategies for health promotion and particularly for coronary heart disease reduction. PMID- 10466375 TI - [The genetic contribution to coronary disease. The genetic factors related to lipid metabolism]. AB - The increasing development of molecular genetics, the progress of the Human Genome Project, and the widespread application of its new methods and molecular techniques provide a new perception of coronary heart disease and a better recognition of genetic markers (mutations and polymorphism) related to new or traditional cardiovascular risk factors. An example of this has been the major effort made over the last years to evaluate and establish the genetic molecular mechanisms that are the basis of the synthesis of apolipoproteins, lipoprotein processing enzymes and lipoprotein receptors. These are some of the subjects discussed in this review [apoB, apoE, atherogenic lipid profile and CETP, and Lp(a)], in which the role of polymorphic alleles and isoforms in cardiovascular risk and coronary heart disease is stressed. PMID- 10466376 TI - [Stents and balloon mitral valvuloplasty]. PMID- 10466377 TI - [Complete coronary revascularization. Reality or myth?]. PMID- 10466378 TI - [The humanization of health care--the psychological aspects of nurses in cardiac surgery]. AB - This paper arose from the need felt, during the position of head nurse of an intensive care unit, to also provide care for nursing staff. A head nurse who promotes both personal and professional development of the team must be aware of the stressful experiences of the nursing staff and attempt to reduce the effect that these experiences may have on them. The sharing of emotions as well as the nurses's need to express feelings, should be promoted and stimulated by the head nurse. Emotions expressed by nurses caring for patients facing death, pain and suffering are a constant concern for a head nurse who desires both the personal and professional development for her staff, and the best possible recovery for the patients in their care. PMID- 10466379 TI - [Pulmonary embolism associated with a crossed embolism with visualization of the thrombus in transit through the foramen ovale]. PMID- 10466380 TI - [The gallery of presidents. Jacinto Moniz de Bettencourt, 4th president of the S.P.C. Sociedade Portuguesa de Cardiologia]. PMID- 10466381 TI - [The results of a survey of cardiology service heads and of the regular members of the Sociedade Portuguesa de Cardiologia on research in cardiology]. PMID- 10466382 TI - [Angiosarcoma of the liver as a cause of massive hemoperitoneum]. AB - The authors give an account of an uncommon cause of haemoperitoneum--spontaneous rupture of an angiosarcoma-afflicted liver. During the urgent operation haemostatic stitches, partial devascularisation of the liver and tamponade were used. The patient died during the postoperative period due to prolonged haemorrhagic shock and developing multiorgan failure. The authors discuss possible methods of haemostasis of hepatic origin, and emphasize perihepatic tamponade (packing). Failure of the surgery of this case-history is discussed. PMID- 10466383 TI - [Empyema of the thorax]. AB - The authors evaluated the therapeutic procedure and results in patients with empyema of the chest. From December 1996 till June 1998 at the Surgical Clinic of the Third Medical Faculty, Charles University Prague 21 patients with empyema of the chest were hospitalized. The most frequent cause of empyema of the chest was pneumonia in 13 patients (62%). In 16 patients (in 76%) the empyema was classified as the third stage of the disease. Decortication, the most frequent procedure, was performed in 16 patients, incl. three where it was done using videothoracoscopy. Surgical treatment was supplemented by antimicrobial treatment, either monotherapy or a combination of antimicrobial preparations. From the total number one patient died 25 days after thoracotomy and partial decortication with a mesiotheloma of the pleura. The other patients have no signs of relapse of empyema. According to the authors experience the selection of the surgical procedure depends on the stage of the disease. In the authors group decortication by the thoracotomic route was used most frequently. PMID- 10466384 TI - [Gradual postischemic reoxygenation as protection against spinal cord ischemia in an experiment]. AB - The authors attempted reperfusion on the experimental model of spinal ischemia described most frequently in the literature. After double ligature of the aorta- closely beneath the insertion of the subclavian artery and closely above the diaphragm they induced for a period of 40 minutes a state similar to surgery of aneurysms of the thoracoabdominal part of the aorta. in 8 dogs of the control group 7 animals developed paraplegia. In the group with graded postischaemic reoxygenation induced by a change of the ventilation ratio of N2O:O2 none of the dogs was paralyzed after two days survival. The neurological condition evaluated according to Tarlov's score was consistent with the histological finding in the preparations of the spinal cord stained according to Naut. Degenerative changes of interneurons were found mainly in the control group. In the group with graded reoxygenation they were rare. This method confirmed the importance of the reperfusion stage for postischaemic damage of the spinal cord. PMID- 10466385 TI - [Epidural regional hypothermia for prevention of paraplegia in experimental aortic clamping]. AB - In experiments on dogs the authors tried to elaborate methods of protection of the spinal cord from ischaemia during surgery of an aneurysm in the thoracolumbal portion of the aorta. They used the model described in the literature with double ligature of the aorta closely below the insertion of the left-sided subclavian artery and closely above the diaphragm for a period fo 40 minutes. In the control group of 8 dogs they observed severe neurological deficiency manifested by paraplegia in 7 animals. In the second group of 8 dogs the authors used local hypothermia of the spinal cord. Hypothermia was produced by epidural administration of 5 degrees C saline which they started to administer five minutes before clamping and continued throughout the period of ischaemia. The temperature of the spinal cord dropped to 26.8 degrees C. In this group none of the dogs developed neurological damage. The results were evaluated by recording spinal somatosensory potentials, by monitoring the neurological condition according to Tarlov and by histological examination of the spinal cord. In the conclusion they emphasize that this method can protect the spinal cord from 40 minute ischaemia. PMID- 10466386 TI - [A splenic abscess of unknown etiology]. AB - The authors describe the case-history of a 7-year-old female patient with a lienal abscess of unknown etiology. The patient was treated by classical splenectomy with a very good final outcome. The authors draw attention to the serious character of the disease even nowadays and to contemporary diagnostic and therapeutic possibilities. PMID- 10466387 TI - [Pleural exudate as a complication of inflammatory pancreatic disease]. AB - The authors present an account on six patients with a pleural exudate associated with inflammatory pancreatic disease, treated during the six-year period from 1992-1998. The authors succeeded after two-week conservative treatment which involved complete parenteral nutrition and repeated thoracocenteses that the pleural exudate disappeared in five patients with chronic pancreatitis. In one instance, a female patient with haemorrhagic necrotic pancreatitis, the bilateral exudate disappeared only after surgical removal of the necrotic portions of the pancreas. The authors discuss diagnostic and therapeutic possibilities of treatment of these conditions. PMID- 10466388 TI - [Spontaneous pneumothorax. Actual aspects]. AB - Based on a group of 77 patients with 93 attacks of spontaneous pneumothorax the authors submit some unconventional views on this entity and recommend an algorithm of therapy. They assume that the "gold standard" of treatment is tubular drainage of the chest, and only if it fails, surgery is the method of choice. PMID- 10466389 TI - [Congenital diaphragmatic hernia manifesting after the neonatal period]. AB - In the course of 16 years at the authors department 46 children with congenital malformation of the diaphragm were operated. This group comprised 12 children (26%) where the first symptoms developed only after the neonatal period. Most frequently the symptoms were repeated respiratory infections (in 9 children), and gastrointestinal complaints in 4 children. The children were operated electively: in 8 patients a left-sided Bochdalek hernia (1x with a sac) was involved, 3 children had a hiatus hernia and one child a Morgagni hernia. After surgery no early or late complications were recorded. In the discussion the authors deal with inborn diaphragmatic hernia with a late manifestation which is mainly a diagnostic problem. Therefore, based on their own experience and consistent with data in the literature, the authors recommend careful aimed examination of children with chronic non-specific respiratory or gastrointestinal complaints. PMID- 10466390 TI - [Reconstruction of the venous outflow in liver transplantation using the "piggyback" technique]. AB - The piggyback technique in venous outflow tract reconstruction has been proposed as an alternative to the conventional technique in liver transplantation. Between November 1994 and November 1997 this technique was used in 29 patients in our institute. Most of the patients were operated on without veno-venous bypass and we observed no complications concerning venous outflow tract reconstruction and no renal failure. Hemodynamic measurements during the operation showed only an insignificant decrease in cardiac index during the partial clamp on the inferior vena cava. The piggyback technique proved to be a convenient and safe approach to the venous outflow tract reconstruction in a majority of liver transplantations and has been used routinely in our institute. PMID- 10466391 TI - [Biliary ileus. Diagnosis and therapy]. AB - Based on retrospective evaluation of therapeutic results of all patients operated at the Second Surgical Clinic of the Second Medical Faculty Charles University on account of biliary ileus during the last 10 years the authors submit their ideas on treatment of this relatively rare disease. Decisive for the patient is early, though frequently difficult assessment of the diagnosis and the speediest possible removal of the obstruction. Enterlithotomy with thorough revision of the gut above the obstruction and removal of the contents is a sufficiently effective procedure. Subsequent elective cholecystectomy is indicated only in patients with further symptoms of biliary disease and confirmed cholecystolithiasis. PMID- 10466392 TI - [The first lung transplantation in the Czech Republic]. PMID- 10466393 TI - [Long-term results of ileo-rectal anastomosis in familial polyposis]. AB - The authors present their experience with 93 patients operated at the First Surgical Clinic of the General Faculty Hospital and First Medical Faculty, Charles University Prague on account of familial adenomatous polyposis (FAP) assembled during 36 year starting in 1962. They analyze 91 patients followed up in collaboration with the First Medical Clinic of the General Faculty Hospital and First Medical Clinic Charles University Prague. Seventy-two of the patients were operated and in 55 of them an ileorectoanastomosis was made following subtotal colectomy. Two important findings were made. From the group of 91 patients incl. primary patients who suffered already from advanced malignant disease of the large bowel a total of 38.5% died. In the rectal stump after ileorectoanastomosis on average within 16 years after operation in 16.4% of the patients a malignant tumour was found. This leads to the belief that patients should be recommended colectomy with ileoanoanastomosis with an ileal reservoir. This operation was performed during the last five years in nine patients with this condition, using a one-stage or two-stage procedure with temporary ileostomy. PMID- 10466394 TI - [Incarcerated Waldeyer's hernia as the cause of necrosis of the small intestine (case report)]. AB - The authors present a case-record of a rare incarcerated Waldeyer hernia which became the cause of infarsation and necrosis of the free portion of the small intestine in the abdominal cavity, which developed as a result of compression of the upper mesenteric vein by the contents of the hernial sac. The authors draw attention to the asset of CT examination for preoperative diagnosis. US examination failed. They explain why, although a rare acute ileous abdomen is involved, the X-ray picture of levels on the native X-ray of the abdomen is lacking. PMID- 10466395 TI - [Treatment of chronic subdural hematoma with a closed drainage system]. AB - The authors evaluate a group of 92 patients with the diagnosis of chronic subdural haematoma confirmed by CT. All patients were operated from two trepanation bores with insertion of drainage into the subdural space and its connection with the closed drainage system. They used three types of drainage systems. The best clinical results were obtained with the negative pressure suction system of Braun Co in 92%. This system was switched on intermittently only till the patient developed signs of headache. Attention is focused on possible types of complications in different systems. In the remaining 13 patients who did not improve reinsertion of the drain was used, or craniotomy with membranectomy with a 60% improvement. The mentioned negative pressure drainage system in treatment in chSDH seems most suitable and is associated with the least number of complications. PMID- 10466396 TI - [Continuous loop stitch in single-layer laparotomy suturing]. AB - The authors evaluate their favourable experience with the use of a new absorbable loop suture for one-layer closure laparotomy. The percentage of dehiscences of surgical wounds was lower in the group of patients where Safil Loop suture was used than in the control group. The speed of completing the suture was also greater and the safety of closure of laparotomy was more favourable. PMID- 10466397 TI - [Laparostomy in the treatment of severe intra-abdominal infections]. AB - The objective of the work was to evaluate the results of comprehensive treatment of protracted primary and secondary intraabdominal infection. Retrospective analysis of a group of patients in 1994-1998 using laparostomy, peritoneal lavage, thorough repeated cleansing of the peritoneal cavity, microbiological monitoring with subsequent aimed antibiosis and comprehensive intensive care provide evidence of the rescue of almost 40% liminally curable patients. The results justify the use of comprehensive therapeutic procedures in late stages of serious peritonitis. PMID- 10466398 TI - [Complications of endovascular therapy of abdominal aortic aneurysms. Analysis and possible treatment]. AB - The required result of correctly implemented endovascular treatment is bridging and elimination of the aneurysmatic sac from the blood flow in the aorta. Perfusion of the sac after release of the endovascular prosthesis is one of the most frequent complications of endovascular treatment. The majority of these perfusions is caused by a leak of the endovascular prosthesis at the site of the stented anastomosis, or more rarely by a collateral flow into the aneurysmatic sac or rupture of the endoprosthesis. Perfusion of the sac "endoleak" complicates endovascular treatment in cca 30%. During the postoperative period about half the perfusions recede spontaneously and about half require further treatment. Based on their own experience the author submits suggestions for the classification of leakage of endovascular prostheses with regard to the method of their treatment. PMID- 10466399 TI - [Chylothorax--a rare type of pleural effusion. Case report]. AB - Chylothorax is a rare type of pleural exudate. The diagnosis, assessment of etiology, reflections on the type of treatment, as well as treatment are associated with various pitfalls. The authors present the case-history of a patient with spontaneous chylothorax on the right who was successfully treated by surgery after possible ways of conservative treatment were exhausted. Surgical treatment involved severing and ligature of the thoracic duct. PMID- 10466401 TI - [Comparison of the results of CT examination and surgical findings in blunt injuries of the liver and spleen]. AB - In the course of 24 months (July 1 1996 till June 30, 1998) after CT examination of the abdomen 23 patients were subjected to laparotomy or laparoscopy as injury of the liver or spleen was assumed. Conclusions of CT are in the literature considered one of the necessary prerequisites of possible conservative treatment of the mentioned injuries, their accuracy and reliability is however not unequivocal. On comparison of the CT and surgical finding in the above patients at the time of examination the positive CT finding was correct in 17 of 19 cases, i.e. in 89%. Four patients were on count of clinical signs of intraabdominal injury operated despite a negative CT, in two of them liver injury was found in the obl. lig. falciforme. The overall accuracy was 83%. During the second assessment the overall accuracy increased to 87%. CT, using the A.A.S.T. classification underrated injury on average by 0.45 degree. In the authors' opinion CT of the abdomen is indicated in multiple injuries and severe associated injuries in case of circulatory stability as it makes it possible to examine effectively within a short time also other injured areas. It should always be made, using contrast. PMID- 10466400 TI - [An abdomino-scrotal hydrocele]. AB - Abdominoscrotal hydrocele is encountered rarely in child age. The authors present an account on two patients with abdominoscrotal hydrocele in a three- and fifteen year-old boy. They discuss its development, draw attention to the not very marked clinical manifestations, diagnostic pitfalls and the contribution of sonography to diagnosis. PMID- 10466402 TI - [Personal experience with administration of Smektit (hydrated magnesium aluminum silicate) in the treatment of diarrhea of non-infectious origin and diarrhea after resection of the large intestine]. AB - Smektit was used for treatment of 24 patients, incl. 10 hospitalized with diarrhoea of a non-infectious etiology (postoperative period, non-adherence to the dietetic regime). Microbiological examination involved only examination of the common flora of the large bowel. In the 10 hospitalized patients the complaints receded very rapidly, after a maximum of three days. In the remaining 14 patients who had resections on account of colorectal carcinoma the diarrhoea was in the majority due to reduction of the large bowel and thus reduction of the resorption area. The condition of ten patients became stabilized and the frequency of bowel movements decreased. In the group of four patients after subtotal colectomy only one patient improved. In three patients smektit administration had no effect. In these patients the cause of frequent bowel movements was the extensive reduction of the large bowel. PMID- 10466403 TI - [Intrathoracic occurrence of post-transplantation lymphoproliferation]. AB - In the submitted case-history the authors wished to draw attention to serious complications after transplantation. Posttransplantation lymphoproliferation (PTLP) is a rare complication of organ transplantation, its incidence amounts to some 2% of organ recipients, in combined heart-lung transplantations the incidence is as high as 10%. The prerequisite of lymphproliferations is infection with the Epstein-Barr virus. The virus causes transformation of B lymphocytes and subsequent lymphoproliferation. Immunosuppressive preparations, due to their effect on Tlymphocytes promote this transformation. The decisive imaging method in tumoriform occurrence is high resolution computed tomography. Based on CT examination surgical biopsy is performed with subsequent in situ hybridization which confirms unequivocally the diagnosis of posttransplantation lymphoproliferation. Only on the basis of results of in situ hybridization treatment may be started which involves restriction or discontinuation of immunosuppressive treatment and administration of antiviral preparations. Frequently this treatment fails and must be discontinued on account of a rejection reaction of the organism. The prognosis in untreated forms is adverse. PMID- 10466404 TI - Combinatorial approach of bacteria to antibiotic resistance. AB - In this minireview, we will consider the combinatorial genetics bacteria use to offset antibiotic selective pressure, i.e., resistance gene associations, organization, and regulation. The quantitative and qualitative consequences of this gene shuffling will be discussed in terms of phenotype. For the sake of simplicity, we will concentrate on Gram-positive cocci (enterococci, streptococci, and staphylococci) of medical importance and on antibiotic classes which are therapeutically relevant. The data provided are mainly, if not exclusively, taken from the work carried out in the laboratory, although there are numerous other examples in the literature. PMID- 10466405 TI - Horizontal gene transfers in the environment: natural transformation as a putative process for gene transfers between transgenic plants and microorganisms. AB - Horizontal gene transfers among bacteria, such as natural transformation or conjugation, may have played an important role in bacterial evolution. They are thought to have been involved in promoting genome plasticity which permitted bacteria to adapt very efficiently to any change in their environment and to colonize a wide range of ecosystems. Evidence that some genes were transferred from eukaryotes, and in particular, from plants to bacteria, was obtained from nucleotide and protein sequence analyses. However, numerous factors, including some which are endogenous to the bacterial cells, tend to limit the extent of transfer, particularly among phylogenetically distant organisms. The goal of this paper is to give an overview of the potentials and limits of natural interkingdom gene transfers, with particular focus on prokaryote-originating sequences which fit the nuclear genome of transgenic plants. PMID- 10466406 TI - Recognition specificity of monoclonal antibodies which protect mice against Salmonella typhimurium infection. AB - We used enzyme-linked immunosorbent assay (ELISA), competitive inhibition ELISA, flow cytometry and western immunoblots to study the antigenic specificity of two monoclonal antibodies (mAbs) raised against the cell surface antigens of Salmonella typhimurium. These mAbs (SH6.11 and WB60.4) protect CAF1 (Ity(r)) mice against endotoxemia and mouse typhoid. We found that SH6.11 and WB60.4 recognize Salmonella serogroup B-specific lipopolysaccharide O4 and O5 factors, respectively. These mAbs did not bind to Salmonella serotypes that belong to serogroup A, D1, E4, G2, or R and did not cross-react with other enteric and nonenteric bacterial species. PMID- 10466407 TI - Two genes from the capsule of Aeromonas hydrophila (serogroup O:34) confer serum resistance to Escherichia coli K12 strains. AB - The Escherichia coli DH5alpha strain as well as other K12-derived strains are unable to produce O-specific lipopolysaccharide and are thus rough and serum sensitive. One representative recombinant clone (COS-SR1) containing Aeromonas hydrophila (serogroup O:34) chromosomal DNA conferred serum resistance to E. coli K12 strains. Genetic, biochemical, and immunological studies suggested that the two genes (orf1 and wcaJ) identified in a subclone (pAC-SR9) of COS-SR1 are necessary for the production of the colanic acid capsule at 37 degrees C on E. coli DH5alpha, rendering the strain serum-resistant. A. hydrophila strains from serogroup O:34 are able to produce capsule when they grow both in synthetic medium and in an autolysate of fish viscera. However, defined wcaJ insertion mutants of A. hydrophila 1051-88 (serogroup O:34) are unable to produce capsule on these media. This strongly suggests that both genes belong to the gene cluster responsible for capsule production (wca) of A. hydrophila 1051-88 (serogroup O:34). PMID- 10466408 TI - Beta-lactamase-mediated resistance to extended spectrum cephalosporins among clinical isolates of Pseudomonas aeruginosa. AB - The role of chromosomal cephalosporinases and secondary beta-lactamases in resistance to extended spectrum cephalosporins in clinical isolates of Pseudomonas aeruginosa was investigated. Strains 687, 59, and 58 expressed an inducible chromosomal cephalosporinase, efficiently enhanced with cefoxitin and imipenem. The inducible activity in strain 802 was produced at a moderately elevated basal level and may be involved in resistance to extended spectrum cephalosporins and aztreonam. All strains produced secondary beta-lactamases inhibited by clavulanate: strains 687, 59, and 58 had carbenicillinases with pIs of 5.7 and 5.3. Strain 802 expressed a secondary beta-lactamase of pI 7.6 which may be a novel extended spectrum beta-lactamase different from known enzymes of P. aeruginosa. PMID- 10466409 TI - Re-evaluation of antibiotic and mercury resistance in Escherichia coli populations isolated in 1978 from Amazonian rubber tree tappers and Indians. AB - A study was carried out to assess the stability of antimicrobial susceptibility of wild isolates upon long-term storage using fifty-three Escherichia coli strains isolated in 1978 from feces of healthy children from the Amazon region in Brazil, exposed to low levels of antimicrobial agents, and examined for resistance to mercury and four antibiotics. All of the strains were kept in Lignieres medium at room temperature and were transferred to fresh media four times during this period. Thirty-five out of the 53 strains analyzed in 1978 were viable. Upon recovery, antibiotic and mercury resistance was estimated. All of the 35 strains maintained their original phenotype in a stable fashion, except for one multiresistant strain which became susceptible to kanamycin. Fifty-four percent of the strains exhibited a resistance phenotype, among which 47% had conjugative plasmids. PMID- 10466410 TI - Biodegradation of phenylbenzoate and some of its derivatives by Scedosporium apiospermum. AB - Scedosporium apiospermum, a recently isolated phenol-degrading hyphomycete, was shown to be able to productively utilise the diaryl ester phenylbenzoate as its sole source of carbon and energy. The characterisation of degradation intermediates together with the detection of the corresponding catabolic enzymes in crude extracts enabled us to propose a pathway for the degradation of this diaryl ester. According to our results, an inducible esterase initiated the biodegradation of phenylbenzoate by hydrolysing the ester bond to yield stoichiometric amounts of phenol and benzoate. While phenol was catabolised via catechol and hydroxyhydroquinone, the benzoate was further degraded via the protocatechuate branch of the ortho-pathway. In addition, the fungus utilised p tolylbenzoate and 4-chlorophenylbenzoate by employing similar catabolic sequences. PMID- 10466411 TI - Renal-coloboma syndrome: a multi-system developmental disorder caused by PAX2 mutations. AB - Optic nerve coloboma combined with renal disease, also called renal-coloboma syndrome ( # 120330 in McKusick's Mendelian Inheritance in Man Online, OMIM), a relatively recently characterized syndrome, results from autosomal dominant mutations in the PAX2 gene. Although renal-coloboma syndrome involves both ocular and renal anomalies, some patients are affected with vesico-ureteral reflux (VUR), high frequency hearing loss, central nervous system (CNS) anomalies, and/or genital anomalies, consistent with the expression of PAX2 in these tissues during development. We review here the clinical features of patients with renal coloboma syndrome and PAX2 mutation. We also review the PAX2 mutations that have been reported to date, and discuss the possible effect of PAX2 mutations on normal development. PMID- 10466412 TI - Crossing the blood-brain barrier to central nervous system gene therapy. PMID- 10466413 TI - Molecular mechanisms controlling lung morphogenesis. AB - The complex process of lung formation is determined by the action of numerous genes that influence cell commitment, differentiation, and proliferation. This review summarizes current knowledge of various factors involved in lung morphogenesis correlating their temporal and spatial expression with their proposed functions at various times during the developmental process. Rapid progress in understanding the pathways involved in lung morphogenesis will likely provide the framework with which to elucidate the mechanisms contributing to lung malformations and the pathogenesis of genetic and acquired lung diseases. PMID- 10466414 TI - Family physicians' perspectives on genetics and the human genome project. AB - The objective of the study was to determine family physicians' attitudes and beliefs about human genetics research and the human genome project (HGP). The design of the study involved qualitative, semi-structured interviews. Primary variables of interest included family physicians' training; their attitudes about the HGP; requests for genetics counseling; and their approaches to counseling requests. The setting was a medium-sized, Midwest, US city. The participants were 16 university-affiliated, community-based family physicians. For contents analysis, we used a coding scheme to identify illustrative themes and subthemes. While most of the family physicians reported familiarity with genetics and the HGP, and experiences with counseling requests, nearly all (15) reported little training in genetics counseling. Four major themes were identified: 1) impact on clinical care; 2) educational issues relevant to genetics and the HGP; 3) ethical concerns; and 4) family medicine responsibilities. These family physicians do not perceive genetics as having a substantial impact on their practice, but do expect major clinical changes in the future. Many feel there have been inadequate educational opportunities to learn about genetics, and some indicate reluctance to invest in self-education until genetic problems become more clinically relevant. These practitioners envision a role for family medicine the specialty to shape priorities in genetics research. PMID- 10466415 TI - Tricho-dento-osseous syndrome and amelogenesis imperfecta with taurodontism are genetically distinct conditions. AB - Amelogenesis imperfecta of the hypomaturation-hypoplasia type with taurodontism (AIHHT) is inherited as a highly penetrant autosomal dominant trait. These dental findings are similar to those of another autosomal dominant condition, the tricho dento-osseous syndrome (TDO), from which AIHHT differs primarily by lack of changes in the hair and bones. TDO is characterized by a highly variable clinical phenotype. While enamel hypoplasia and taurodontism appear to be present in all TDO cases, non-dental features may be absent, with approximately half of TDO cases losing the kinky/curly hair phenotype seen in infancy by adolescence, and in almost 20% of cases, osseous changes are not evident. The genetic basis for AIHHT is unknown and it has been questioned whether AIHHT and TDO are separate conditions or a spectrum of disease. The genetic basis for TDO has recently been identified as a deletion mutation in the distal-less 3 (DLX3) transcription factor gene. To determine if AIHHT and TDO represent variable expression of a common DLX3 gene mutation, allelic mutations of the DLX3 gene, or mutations in DLX7 (the linked paralogue to DLX3 on chromosome 17), we have performed mutational analysis and sequencing studies of the DLX3 and DLX7 genes in three individuals (two affected and one unaffected) from a family with AIHHT. Results of the analysis demonstrate that AIHHT and TDO are not due to a common DLX3 gene mutation. Sequence analyses of the DLX3 and DLX7 genes suggest AIHHT is not due to genetic mutations or polymorphisms in the exons of these genes. These results suggest that AI-HHT and TDO are two genetically distinct conditions. PMID- 10466416 TI - Cohen syndrome: evaluation of its cardiac, endocrine and radiological features. AB - Cohen syndrome (MIM no. 216550) is an autosomal recessive disorder with a typical clinical picture. Since the first report, most publications have represented single case reports. In this study, our aim was to describe cardiac, endocrine and radiological abnormalities in 22 Cohen patients of Finnish descent. Detailed investigations of the heart revealed the anatomy of the heart to be normal with no evidence for clinically significant mitral prolapse. However, a decreased left ventricular function with advancing age was identified. No significant endocrine abnormalities were found at the examination of pituitary, adrenal and thyroid function. The height was either normal or patients were moderately short (mean height standard deviation score (SDS) - 2) at all ages, associated, however, often with the marked kyphosis. Truncal obesity was seen in 4/22 patients. X-rays of the chest, lumbar and thoracic spine, long bones, ankles and metacarpophalangeal pattern profiles revealed kyphosis, scoliosis and calcaneo planovalgus as common features. Fingers of these patients were slender but short with a characteristic metacarpophalangeal pattern profile. PMID- 10466417 TI - Epidemiology of dystrophinopathies in North-West Tuscany: a molecular genetics based revisitation. AB - A molecular genetics-based epidemiological investigation was carried out in 1997 in the territory of North-West Tuscany, central Italy, to calculate incidence and prevalence rates of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD). Results were compared with a previous epidemiological study conducted in the same area in 1981, in the pre-dystrophin era. Routine adoption of methods of molecular diagnosis determined an increase in prevalence of BMD from 1.06 x 10(-5) to 2.42 x 10(-5) inhabitants, while cumulative incidence of DMD was markedly decreased from 23.12 x 10(-5) during the period 1965-1976 to 10.71 x 10(-5) male live births during the period 1977 1994. The combined reduction of DMD/BMD diagnostic error rate and familial recurrence could explain these results, providing the bases for a consistent redefinition of dystrophinopathy carrier frequency in the area considered. PMID- 10466418 TI - High frequency of tissue-specific mosaicism in Turner syndrome patients. AB - Interphase fluorescent studies of X chromosome aneuploidy in cultured and uncultured blood lymphocytes and oral mucosa epithelial cells using X centromere specific DNA probe in addition to standard karyotype analysis were performed in 50 females with a clinical suspicion of Turner syndrome. All the patients were previously screened for the presence of 'hidden' Y chromosome mosaicism, using the primers DYZ3 and DYZ. The use of fluorescence in situ hybridization (FISH) analysis of interphase nuclei of tissues from different germ layers (lymphocytes from mesoderm and buccal epithelial cells from ectoderm) improves the accuracy of detection of low-level mosaicism. FISH studies on interphase nuclei revealed that 29% of patients with a pure form of monosomy X detected by metaphase analysis are, in fact, mosaics. The level of cells with the normal chromosomal constitution in lymphocytes of these cases as a rule was low, ranging from 3 to 18%, with an average of 7%. Two false-positive cases and one false-negative case of X monosomy mosaicism determined by standard cytogenetic approach were detected using FISH analysis. The majority of patients (92%) with mosaic form of Turner syndrome have considerable tissue-specific differences in levels of X aneuploidy. Our data indicate that in cases when mosaic aneuploidy with low-level frequency is questionable (approximately 10% and lower), the results of standard metaphase analysis should be supplemented with additional FISH studies of interphase nuclei. Tissue-specific differences in contents of different cell lines in the same patients point to the necessity of studying more than one tissue from each patient. PMID- 10466419 TI - Mucopolysaccharidosis type I: characterization of novel mutations affecting alpha L-iduronidase activity. AB - alpha-L-Iduronidase (IDUA) deficiency (mucopolysaccharidosis type I, MPS I) involves a broad spectrum of clinical severity ranging from a severe Hurler syndrome through an intermediate Hurler Scheie syndrome to a mild Scheie syndrome. To date, a number of mutations of the IDUA gene are known in Hurler syndrome, but only a few in Hurler Scheie or Scheie syndrome. The characterization of novel mutations in two patients with the Hurler-Scheie syndrome is reported on. The novel R619G mutation (C-G transversion in codon 619) was apparently homozygous. In transfected COS-7 cells, R619G caused significant reduction in enzyme activity (1.5% of normal activity), although it did not cause significant reduction in IDUA mRNA or protein level. Conversely, the previously described homozygous T364M mutation (C-T transition in codon 364) caused a decrease in the level of IDUA mRNA. Studies inhibiting RNA synthesis with actinomycin D or inhibiting protein synthesis with cycloheximide demonstrate that the decrease in the latter mutation is attributable to an increased rate of mRNA decay. By examining the stability of IDUA mRNA and protein, studies provide better insight into the effect of mutation on IDUA activity. PMID- 10466420 TI - Homozygosity for a novel DTDST mutation in a child with a 'broad bone platyspondylic' variant of diastrophic dysplasia. AB - Atypical or variant forms of well-known chondrodysplasias may pose diagnostic problems. We report on a girl with clinical features suggesting diastrophic dysplasia but with unusual radiographic features including severe platyspondyly, wide metaphyses, and fibular overgrowth, which are partially reminiscent of metatropic dysplasia. The diagnosis was clarified by molecular analysis of the DTDST gene, which revealed homozygosity for a previously undescribed mutation leading to a Q454P substitution in the 10th transmembrane domain of the DTDST sulfate transporter. Molecular analysis may be of particular value in such atypical cases. PMID- 10466421 TI - Germline mosaicism in X-linked myotubular myopathy. AB - X-linked myotubular myopathy (XLMTM; OMIM310400) is a congenital muscle disorder characterized by severe hypotonia and respiratory insufficiency. The disorder was mapped to Xq28 by linkage studies and the MTM1 gene was isolated by positional cloning. The gene product is a 603 amino acid protein named myotubularin. A small domain in its sequence shows high homology to a consensus active site of tyrosine phosphatases, a diverse class of proteins involved in signal transduction, control of cell growth, and differentiation. In this report, two brothers affected with XLMTM are shown to have a point mutation (G1187A) in exon 11 of the MTM1 gene. Surprisingly, their mother does not have this mutation in her lymphocytes. Therefore, she likely has a germline mosaicism. As this is the third report of germline mosaicism in XLMTM, the finding has important implications for genetic counseling. PMID- 10466422 TI - A locus for autosomal recessive achromatopsia on human chromosome 8q. AB - Autosomal recessive achromatopsia is a rare disorder characterized by total absent color vision, nystagmus, photophobia, and visual impairment, frequently leading to 'legal blindness'. The primary defect is at the photoreceptor level, with retinal cones being absent or defective. The first locus for this disorder was mapped to chromosome 2q11. Here, we confirm the genetic mapping of a locus discovered in our studies of a kindred with Irish ancestry, but no known consanguinity, in which 5 of 12 children are affected. We have mapped the locus in this disorder in this family to chromosome 8q. Available data now narrow the region containing the putative gene to 1.2 cM. PMID- 10466423 TI - A de novo complex chromosomal rearrangement with nine breakpoints characterized by FISH in a boy with mild mental retardation, developmental delay, short stature and microcephaly. AB - A de novo complex chromosome rearrangement (CCR) involving chromosomes 1, 6, 7, 15 and Y was detected in a boy with mental retardation, short stature, and microcephaly. Fluorescence in situ hybridisation (FISH) with whole chromosome painting libraries, band-specific cosmids and telomeric probes was essential for the characterisation of the rearrangement. The CCR was shown to be the result of at least nine chromosomal breaks and involved the alternating insertion of two segments of the short arm of chromosome 1 and two segments of the long arm of chromosome 6 into a novel derived chromosome 7. A non-reciprocal translocation between the distal short arm of the same chromosome 7 and the distal long arm of the Y chromosome was also found, together with a paracentric inversion of the long arm of chromosome 15. The only detectable imbalance was a deletion of the heterochromatic Yq telomeric region. FISH investigations in this case have revealed an additional complexity in this CCR, which has implications for reproductive risk assessment and genetic counselling. PMID- 10466424 TI - Infantile spasms in a patient with partial duplication of chromosome 2p. PMID- 10466425 TI - Distal arthrogryposis type IIB: probable autosomal recessive inheritance. PMID- 10466426 TI - Screening for FMR1 mutations among the mentally retarded: prevalence of the fragile X syndrome in Spain. PMID- 10466427 TI - Homozygosity for two mild glucocerebrosidase mutations of probable Iberian origin. PMID- 10466428 TI - Triple aneuploidy in spontaneous abortions. PMID- 10466429 TI - About the antiemetic effectivity of granisetron in chemotherapy-induced acute emesis: a comparison of results with intravenous and oral dosing. AB - The paper is devoted to review literature data on intravenous and oral antiemetic effectivity of granisetron (GRAN) a selective 5HT3 antagonist and to determine the optimal dose for the prophylaxis of chemotherapy-induced acute emesis. The drug was put on the market in the injectable form in 1994 and in the oral form in 1995, so a sufficient number of reports have been published for the evaluation. According to the summarized data on 6095 patients treated with intravenous GRAN, on average 66% antiemetic complete response (CR) rate was reached (i.e. no vomiting in the first 24 hours of chemotherapy). Best results were observed with the dose of 40 microg/kg intravenous GRAN, on average 70% CR (range 47-93%) were achieved in 4182 of the 6095 patients with this dose. In 942 patients treated with the mostly applied oral dose of GRAN (1 mg twice daily), on average 61% CR (range 52-82%) were reported. Side effects were weak and transient, mostly headache and constipation were observed. Headache appeared in 13% with the use of 40 microg/kg intravenous dose GRAN and in 18% with 1 mg oral dose twice daily Constipation was observed in 5.3% and 17% with the injection and oral dose, respectively. PMID- 10466430 TI - FASAY: a simple functional assay in yeast for identification of p53 mutation in tumors. AB - Alteration of the p53 tumor suppressor gene is the most common genetic defect known to occur in human tumors. Germ-line p53 mutations significantly increase the risk of developing diverse malignancies. FASAY is a simple functional assay for germ-line and somatic mutations in the p53 gene altering the transactivation capability of the p53 protein. The method was successfully used for mutation analysis of p53 in various cell lines, somatic tumor cells and blood cells. In addition, FASAY was also found effective as a tool for basic research of binding of mutant p53 proteins to promoters of different p53 target genes. PMID- 10466431 TI - Kojic acid--a new leading molecule for a preparation of compounds with an anti neoplastic potential. AB - Kojic acid as a molecule of natural origin may serve as template for the synthesis of new biologically active compounds. The synthetic KA (pyranone) derivatives possess various kinds of biological activities which are related by their similarity to flavonoids. The most important property is the antifungal and antineoplastic activity and capability of chelating metals. It is shown that the antineoplastic activity of kojic acid derivatives is based on various mechanisms of action on different levels of cellular metabolism and functions what makes this compound interesting for future investigation as cytotoxic agent. PMID- 10466432 TI - Symptom pattern and diagnostic work-up of malignancy at first symptom presentation as related to level of care. A retrospective study from the primary health care centre area of Kungsbacka, Sweden. AB - This retrospective study was aimed to characterize the diagnostic process of cancer with respect to level of care, initial symptoms, and diagnostic procedures. It was based on analysis of medical records of all subjects with colorectal, pulmonary, breast or prostate cancer, reported to the Swedish Cancer Registry during defined periods of time in the community of Kungsbacka with about 46,500 inhabitants. Initial symptoms, diagnostic procedures, outcome of diagnostic procedures, level of care, and doctor's delay were analyzed. Most patients (62-73% for the different cancers studied) first visited a general practitioner for the symptoms which lead to the diagnosis of cancer. The most common initial symptom for colorectal cancer was defecation abnormality, for breast cancer a palpable mass in the breast, for pulmonary cancer cough, and for prostate cancer symptoms of prostatism. There was no difference in doctor's delay between general practitioners and other physicians. Nonspecific blood laboratory tests made little contribution to the diagnosis of cancer. The results indicate that most cancers of the types studied are diagnosed in primary health care and that it is possible to improve the identification of the few malignant cases among the "noise" of benign diseases, both with respect to accuracy and cost effectiveness. It seems that focused investigations such as fecal occult blood tests and rectoscopy should be more frequently used in patients with gastrointestinal symptoms. PMID- 10466433 TI - Copper, zinc and superoxide dismutase in precancerous, benign diseases and gastric, colorectal and breast cancer. AB - The aim of the present study was to assess serum levels of copper and zinc levels and erythrocytes Cu,Zn-SOD activity and to determine probable changes in gastric and colorectal precancerous diseases, benign breast diseases, gastric, colorectal and breast cancer. The study included 165 subjects with cancer, 348 subjects with precancerous (atrophic gastritis, gastric adenoma, colon adenoma, rectal adenoma) and/or benign diseases (weak dysplasia, severe dysplasia, fibroadenoma, cystic disease) and 161 randomly selected healthy controls. Our results suggest that while in gastric and colorectal cancer there were mostly increased copper levels, in breast cancer they were not changed. Zinc levels were weakly decreased in atrophic gastritis, gastric adenoma and breast cancer. There was a strong positive correlation between zinc levels and SOD activity in fibroadenoma and a weak positive correlation in colorectal adenoma and colorectal cancer without any correlation between SOD activity and copper in these groups. In gastric precancerous disease there was a positive correlation between SOD and copper. The results of this study suggest that serum trace element levels and activity of related enzymes might be different in various neoplastic processes. This variation in neoplastic processes might be influenced by other factors that have to be considered in complex relationships between the whole body and neoplastic cells. PMID- 10466434 TI - Inhibitory effect of Trianthema portulacastrum L. diethylnitroso-amine-induced phenobarbital promoted hepatocarcinogenesis. AB - The chloroform extract (active components from the exotic weed Trianthema portulacastrum L. of Aizoaceae, showed remarkable effect in the reduction of DENA induced rat hepatocarcinogenesis. Hepatocarcinogenesis was induced in male Sprague-Dawley rats by a single intraperitonial injection of diethylnitrosoamine (DENA) at a dose of 200 mg/kg body weight. Chloroform extract of T. portulacastrum at a dose of 100 mg/kg/body wt. to the basal medium/per animal once daily was administered following DENA injection. Morphometric evaluation revealed that the fraction reduced the incidence, numerical preponderance, multiplicity and size distribution of visible pre-neoplastic nodules. Further focal lesions showed a reduction of altered liver cell foci/cm2 and a reduction of average focal area. A decrease in the percentage of liver parenchyma occupied by foci all seem to suggest the anticarcinogenic potential of Trianthema portulacastrum in DENA-induced rat hepatocarcinogenesis. PMID- 10466435 TI - Estradiol receptor and prognostic parameters of human breast cancer. AB - Estradiol receptors are regarded to predict a likely success of hormonal therapeutic efforts and the prognosis of breast cancer patients. But today its prognostic importance is controversial, discussed as either reflecting intrinsic property of the tumor tissue or better therapeutic accessibility of receptor positive tumors. Moreover, the most important clinical prognosticators--tumor size and axillary lymph node involvement do not seem to be related to the estradiol receptor status. In our investigation, the length of disease free interval is similar in estradiol receptor positive and negative patients and in all sites of distant metastases, but it is significantly reduced if more than 4 axillary lymph nodes are involved. Post recurrence survival is significantly longer in estradiol receptor positive than negative patients and also in patients treated by tamoxifen containing therapies. Its length is independent of the number of axillary lymph node metastases and the type of distant metastases, with a tendency to be longer in estradiol receptor positive than negative patients. In addition, the overall survival is longer for estradiol receptor positive than negative patients and becomes reduced with more than 4 axillary lymph node metastases. Frequency of deaths in estradiol receptor positive patients is half that of negative subjects. Furthermore, the length of overall survival is independent on the type of distant metastases, with tendency to be longer in estradiol receptor positive than negative patients. Longest overall survival could be observed for estradiol receptor positive patients who got therapy regimens containing tamoxifen. The weak prognostic advantages of estradiol receptor positive patients are interpreted by estradiol receptors as intrinsic parameters of breast cancer tissue characterizing more its biological behavior than therapeutic accessibility. PMID- 10466436 TI - Enhanced activity of estramustine, vinblastine, etoposide, and suramin in prostate carcinoma. AB - Following hormonal therapy, few treatment regimens have activity in metastatic prostate cancer. Cytotoxic agents have minimal activity in this disease. However, combinations of cytotoxic agents may be beneficial. The activity of estramustine, vinblastine, etoposide, and suramin on cell growth was evaluated. Prostate specific antigen (PSA) is routinely used as a surrogate marker for disease progression. Many pharmacological agents alter PSA levels independently of their effect on tumor growth, the effect of these agents on PSA secretion was determined. Each agent was evaluated alone and in combination with the other drugs in two prostate cancer cell lines. In LNCaP cells, estramustine and suramin were cytostatic, while vinblastine and etoposide were cytotoxic. Estramustine down-regulated etoposide PSA secretion, while suramin had no effect. The effects of etoposide and vinblastine on PSA secretion were not evaluable. In PC-3 cells, only etoposide was cytotoxic. Tandem combinations were more cytotoxic than single agents in both cell lines. The addition of a third agent to the tandem combination produced less cytotoxicity. In our hands, the best combinations were estramustine/vinblastine, suramin/vinblastine, and suramin/etoposide. These combinations yielded 20-60% higher cytotoxicity than any of the drugs alone. PMID- 10466437 TI - The surgical treatment of adrenal gland tumors--incidentaloma. AB - Analysis of 32 patients operated on because of accidentally discovered adrenal tumors "incidentaloma" is presented. In 12 of them there was subclinical hormonal activity, in 9 of them tumors turned out to be pheochromocytoma and 3 of them were cortex adenoma. There were 20 hormonally inactive tumors, in 5 of them there were malignant lesions (4 of the cortex and 1 of the medulla). For evaluation of hormonal activity of adrenal tumors evaluation of chromogranin A and cortisol serum blood level or urine free cortisol level is recommended. For precise localization of the tumor beside USG also CT examination is of use. According to the high percentage of malignant lesions in "incidentaloma" type tumors, surgery treatment without delay is recommended. BAC or DHES in blood serum examinations were not found helpful in preoperative evaluating the lesions as benign or malignant. In case of preoperatively found subclinical hormonal hyperactivity of medulla pharmacological treatment with alpha and beta blockers in surgery preparation is recommended. Lateral extraperitoneal access for adrenalectomy is considered safe and provides good operational view. Laparoscopic procedure because of high percentage of malignant lesions in this group of patients is not justified. PMID- 10466438 TI - Primary cancer of the fallopian tube with transitional differentiation. Clinical and pathological assessment of 6 cases. AB - To establish prognosis, histologic appearance and p53 and c-erbB-2 expression in cancer tissue, six cases of primary transitional cancer of the fallopian tube were analyzed. Among 45 patients with the primary cancer of the fallopian tube diagnosed between 1992 and 1997, we found six cases diagnosed previously as solid (undifferentiated) cancer of the tube. p53 protein and c-erbB-2 oncoprotein expression were examined using an avidin-biothinyl-peroxydase complex method. The accumulation of p-53 protein and c-erbB-2 oncoprotein were used as prognostic marker of the transitional cancer of the tube. According to histologic picture all patients were diagnosed for primary cancer of the tube with transitional differentiation. In 4 cases strong positive and in 2 cases moderate positive reaction with antibody against p53 protein was seen for p53 protein. No positive expression of c-erbB-2 oncoprotein in membrane of cancer cells in our cases was detected. Presence p-53 protein in all our 6 cases deny the usefulness of the p53 protein as prognostic marker in primary cancer of the fallopian tube. Lack of expression of c-erbB-2 oncoprotein in membrane of cancer cells is significantly contributed to better prognosis in cases with primary cancer of the tube with transitional differentiation. PMID- 10466439 TI - The role of stable disease in objective response assessment and its impact on survival in advanced colorectal cancer: is "stable disease" a homogenous response category? AB - Stable disease is a category which is not included in the evaluation of the overall treatment response rate. In many studies with a response rate below 20%, chemotherapy almost doubles the survival of patients. In the most chemotherapy trials with advanced colorectal cancer patients, about 30-50% had stable disease. Despite belonging to the same category of therapy response, some patients with stable disease have achieved symptom improvement, but some have not. The aim of the study was to investigate whether the stabilization of the disease with clinical benefit is associated with benefit in survival. A total of 99 patients with advanced colorectal cancer were treated with carboplatin (80 mg/m2, day 1 7), 5-FU (750 mg/m2, day 1-5), leucovorin (100 mg/m2, day 1-5) every 4 weeks. After 4 courses, in the case of stable disease (SD), the patients were stratified according to clinical benefit achievement in: Group A--patients with clinical benefit who continued with chemotherapy until 8 cycles or until disease progression; group B--patients without clinical benefit in whom chemotherapy was stopped after 4 cycles. Clinical benefit was a composite of assessment of pain, ECOG performance status, weight and temperature. Clinical benefit required a sustained improvement in at least one parameter without worsening in any other. Of 97 evaluable patients 48 achieved stable disease. Of 22 pts. with SD clinical benefit performance status improvement was recorded in 17, pain relief in 14, improvement in body weight in 14 and temperature disappearance in 8 pts. Of 26 pts. with SD without clinical benefit, 7 were asymptomatic from beginning of the chemotherapy. No difference was detected in the survival between responders and SD clinical benefit pts. (p = 0.24), but there was significant difference between responders and SD pts. without clinical benefit (p = 0.0004). SD clinical benefit pts. had significant difference in survival in comparison to pts. with progressive disease (p = 5.1 x 10(-6)). The results of our study indicate that under category "stable disease" there are two different subpopulations of patients with quite different symptom response to chemotherapy, different time to progression and possible different survival. PMID- 10466440 TI - Prevalence of chronic idiopathic neutropenia of adults among an apparently healthy population living on the island of Crete. AB - The aim of the present study was to investigate the prevalence of chronic idiopathic neutropenia of adults (CINA) among an apparently healthy population born and living on the island of Crete. The study was carried out with 778 subjects, 392 men aged 16-78 years (median 43 years) and 386 women aged 15-79 years (median 40 years). All were employees of the Medical School or the adjacent University hospital and members of their families. Among these there were 64 subjects (8.23%) who fulfilled the diagnostic criteria of CINA applied in our department. Mild neutropenia (neutrophils 1700-2499/microl) accounted for 6.81% and moderate neutropenia (neutrophils 600-1699/microl) for the remaining 1.41%. No cases of CINA with severe neutropenia (neutrophils below 600/microl) were found. CINA was more frequent in women, with a women to men ratio of about 3:2. Approximately two thirds of the cases appeared in patients aged 30-59 years. Concomitant thrombocytopenia was found in three of the 64 subjects with CINA. Neutropenic subjects had chronic (perennial) rhinitis 3.4 times more frequently than non-neutropenics. No influence of occupation, use of insecticides and pesticides, contact with industrial chemicals, or administration of nonsteroidal anti-inflammatory drugs on the development of CINA was documented. We conclude that, despite the biased character of the study (population not randomly selected), our data provide a valuable estimation of the prevalence of CINA in the general population, given that our sample was sufficiently large, was derived from all major regions of the island, and was composed of subjects of both genders and of all age-groups from 15 to 79 years. PMID- 10466441 TI - Granulocyte colony-stimulating factor-supported combined immunosuppressive therapy (antilymphocyte globulin, cyclosporine, and methylprednisolone) in patients with aplastic anemia: tolerability, efficacy, and changes in the progenitor cell compartment. AB - Neutropenic infections are the major cause of morbidity and mortality in the treatment of aplastic anemia (AA) with antilymphocyte globulin (ALG), cyclosporin A (CSA), and methylprednisolone (MP). Recent data suggest a beneficial effect of administering G-CSF as an adjunct to immunosuppression. We have treated 11 consecutive patients with AA using a combined immunosuppressive regimen including ALG, CSA, and MP plus G-CSF at a dose of 5 microg/kg/day until neutropoietic recovery. In addition to measuring routine hematological parameters we have performed serial determinations of reticulocyte counts and in vitro progenitor cell cultures before and after therapy in order to assess their predictive value for treatment response and to determine the impact of therapy on early hematopoiesis. One patient died on day 34 of neutropenic septicemia. At 1 year, 81% of patients showed response to treatment. The median time to ANC values >0.5 and >1.0 x 10(9)/l were 19 and 35 days, respectively. Reticulocyte counts started to recover after 6 weeks, and transfusion independence was observed on day 52 for red blood cell transfusions and on day 53 for platelet concentrates. All patients with detectable colony formation in peripheral blood achieved a complete hematological remission, as compared with only one of five patients without progenitor cell growth. Although normal ranges were rarely achieved, there was a small but definitive improvement in progenitor cell numbers as compared with baseline values in most patients. Our results confirm the good tolerability and high efficacy of this G-CSF-supported combined immunosuppressive therapy for AA. Detectable colony growth at diagnosis seems to predict a high chance for complete hematological response. PMID- 10466442 TI - Origin of stroma cells in long-term bone marrow cultures from patients with acute myeloid leukemia. AB - Bone marrow stroma cells from patients with acute myeloid leukemia (AML) display a variety of functional abnormalities. In order to determine whether this is related to an imbalance in the proportion of different stroma cell types or to integration of leukemic progeny into the regulatory cell network, stroma layers were established in mycophenolic acid-treated long-term marrow cultures from 16 patients with AML and 42 controls and analyzed by means of simultaneous membrane immunofluorescence and interphase cytogenetics. Macrophages were identified by CD14 expression, fibroblasts by staining with the AS02 antibody, and malignant cells by leukemia-specific numerical chromosome aberrations, including monosomy 7 and trisomy 8. Compared with normal controls, there was a slight decrease in the proportion of stroma fibroblasts (52+/-27% versus 77+/-5%) in 10-week-old cultures from patients with AML. Two of five AML patients with trisomy 8 and both patients with monosomy 7 had evidence of leukemic stroma cells. Most malignant cells were CD14+ macrophages (3.8-98.1% of all CD14+ cells), but some were AS02+ (2.8-5.2%). AML stroma layers showed a reduced capacity to support the growth of normal hematopoietic cells in standard two-stage long-term cultures, but this was unrelated to the presence or absence of leukemic stroma elements. In conclusion, AML populations vary with respect to their ability to produce a malignant microenvironment. Functional defects in the hematopoietic microenvironment, however, are not limited to AML patients with cytogenetically abnormal stroma cells, but extend to cases without evidence of malignant stroma cells. PMID- 10466443 TI - Prevalence of co-morbidity and its relationship to treatment among unselected patients with Hodgkin's disease and non-Hodgkin's lymphoma, 1993-1996. AB - A population-based series of patients with cancer is likely to comprise more patients with serious co-morbidity than clinical trials because of restrictive eligibility criteria for the latter. Since co-morbidity may influence decision making, we studied the age-specific prevalence of co-morbidity and its relationship to applied treatment. Data on all 194 patients with Hodgkin's disease (HD) and on 904 patients with non-Hodgkin's lymphoma (NHL) diagnosed between 1993 and 1996 were derived from the Eindhoven Cancer Registry. In the age group below 60 years, 87% of patients with HD and 80% with NHL did not have a co morbid condition. The prevalence of serious co-morbidity was 56% for patients with Hodgkin's disease who were 60 years and over and 43% and 61% for non Hodgkin patients who were 60-69 years and 70 years and over, respectively. The most common co-morbid conditions were cardiovascular disease (18%), hypertension (13%), chronic obstructive pulmonary disease (COPD; 13%), and diabetes mellitus (10%) for elderly Hodgkin's patients. For non-Hodgkin's patients of 60-69 years and 70 years and over, cardiovascular disease (15 and 22%, respectively), hypertension (14 and 14%, respectively), COPD (6 and 10% respectively), and diabetes mellitus (8 and 10%, respectively) were the most prevalent co-morbid conditions. The presence of co-morbidity was not related to stage or grade of disease at diagnosis. In the presence of co-morbidity, 50% less chemotherapy was administered to elderly patients with Hodgkin's disease and 10-15% less to elderly patients with non-Hodgkin's lymphoma. The presence of co-morbidity was associated with a decreased overall survival within the first 4 months after diagnosis in both Hodgkin's disease and non-Hodgkin's lymphoma for all age groups. In conclusion, serious co-morbidity was found for more than half of all lymphoma patients who were 60 years and older. Elderly patients with serious co morbidity received chemotherapy less often, which is likely to affect survival adversely, as was indicated by a decreased survival within the first 4 months after diagnosis. PMID- 10466444 TI - Chediak-Higashi-Steinbrinck syndrome (CHS) in a 27-year-old woman--effects of G CSF treatment. AB - Chediak-Higashi-Steinbrinck syndrome (CHS) is a rare autosomal recessive disorder which is usually lethal in early childhood. Diagnostic hallmark is the occurrence of giant inclusion bodies in peripheral leukocytes and their bone marrow precursors. We report on a 27-year-old female patient who was admitted for treatment of a skin abscess. She recovered after intravenous antibiotic treatment and surgical incision. Hematological investigation was initiated because of a persisting neutropenia of 15%, with a leukocyte count initially in the normal range but subsequent leukopenia. Case history revealed recurrent skin infections from childhood on, regularly requiring surgical intervention. One year prior to admission a neuropathy had been diagnosed, while a partial albinism had been known for years. Microscopic examinations of peripheral blood and bone marrow aspirate smears were diagnostic for CHS. Additionally, a secondary antibody deficiency was found. Normalization of the white blood cell count, including the differential count, was observed following initiation of G-CSF treatment. Functional assessment of phagocytosis and oxidative burst activity of granulocytes revealed normal results before and after stimulation with G-CSF, however, natural killer cell activity was only weak, with slight improvement after G-CSF treatment in vivo. Cytogenetic analysis showed a normal female karyotype. Although the haploidentical brother of the patient may serve as an allogeneic stem cell donor, transplantation has been postponed because of further deterioration of her already existing CHS-specific neurological impairment. Nevertheless, while receiving G-CSF maintenance treatment our patient experienced no further infectious episodes within 6 months after diagnosis of CHS. PMID- 10466445 TI - Successful treatment with arsenic trioxide of a patient with ATRA-resistant relapse of acute promyelocytic leukemia. AB - Arsenic trioxide has recently been introduced as a promising new agent to treat refractory acute promyelocytic leukemia (APL). In the present study, arsenic trioxide was given intravenously for 42 days to a 56-year-old female patient suffering from chemotherapy/ATRA-resistant APL, with 43% APL blasts in the bone marrow and elevated D-dimers. During the first days of arsenic trioxide treatment a rapid decrease in the D-dimers was seen (normal values reached until day 7), together with a slight decrease in peripheral blood leukocytes. This initial coagulation response was followed by a second phase of hematological response (starting on days 15-20) characterized by leukocytosis, occurrence of myeloid progenitor cells in the peripheral blood, and a decrease in bone marrow blasts (<1% on days 28 and 36). Finally, the patient entered complete hematological and cytogenetic remission, although the PML-RAR alpha fusion product was still detectable by PCR. These data confirm the therapeutic value of arsenic trioxide in relapsed/resistant APL. PMID- 10466446 TI - Pseudotumour cerebri in acute promyelocytic leukemia: improvement despite continued ATRA therapy. PMID- 10466447 TI - Massive disseminated intravascular coagulation and hyperfibrinolysis in alveolar rhabdomyosarcoma: case report and review of the literature. AB - A 68-year-old woman presented with profuse hemorrhage and other signs suggesting an acute leukemia. Histologic and cytogenetic evaluation of her bone marrow revealed alveolar rhabdomyosarcoma as the underlying cause of massive disseminated intravascular coagulation and hyperfibrinolysis. A review of the literature reveals that coagulopathy appears to be a common feature of alveolar rhabdomyosarcoma. PMID- 10466448 TI - Severe bleeding diathesis in a premature baby with extensive hepatic necrosis due to portal vein thrombosis of prenatal onset. PMID- 10466449 TI - Results of conservative treatment of upper urinary tract transitional cell carcinoma. AB - BACKGROUND: The treatment preserving the kidney for upper urinary tract tumor is still controversial. The indications and results of conservative treatment remain to be elucidated. Experiences of this type of treatment are reported. METHODS: Between April 1981 and March 1998, 14 patients with upper urinary tract transitional cell carcinoma were treated with renal preserving methods. Five were elective and nine were imperative cases. Treatments performed were partial nephrectomy, partial ureterctomy with or without adjuvant chemotherapy, endoscopic tumor resection and topical bacillus Calmette-Guerin instillation in one, 10, two and one patient, respectively. RESULTS: Crude and cause-specific 5 year-survival rates were 91.7 and 100%, respectively. Of 14 patients, five had bladder recurrences, but ipsilateral local recurrence developed in only one patient. Two patients died from metastasis of transitional cell carcinoma 61 and 89 months after initial treatment. The lesions of carcinoma in situ were well controlled with topical bacillus Calmette-Guerin therapy. CONCLUSION: The results of conservative treatment for upper urinary tract tumor were satisfactory and local excision can be indicated for low grade, solitary tumors located in the distal ureter. PMID- 10466450 TI - Orthotopic ileal neobladder in male patients: functional outcomes of 66 cases. AB - BACKGROUND: Orthotopic urinary diversion has become the preferred form of bladder reconstruction after cystectomy. We report on our experience with 66 male patients undergoing this procedure from November 1990 to February 1998. METHODS: A neobladder was constructed using an ileal segment with a Hautmann type bladder. Complications were assessed and subdivided into early and late types. Voiding function was evaluated in terms of voiding pattern and continence. Median follow up was 19.5 (range 3.5-87.7) months. RESULTS: There was one (1.5%) perioperative death. The most frequent pouch-related and unrelated early complications were persistent urine leak (7.6%) and prolonged ileus (16.7%), respectively, the majority of cases of which were managed conservatively. Analysis of late complications revealed 6.2% ureteroileal stenosis and 1.5% urethrointestinal stenosis rates, but no case of bladder stone formation. Of the 61 patients in whom voiding function was evaluable, 95.1% achieved excellent daytime continence, while only 67.2% had night-time continence. With regard to posture at voiding, 23 (37.7%) voided in a sitting position. Three of the patients (4.9%) were unable to void and required regular intermittent catheterization. CONCLUSIONS: An orthotopic neobladder can be constructed with acceptable morbidity and excellent functional results. We believe that orthotopic urinary diversion offers an attractive alternative to a bladder substitute when cystectomy is required. PMID- 10466451 TI - Quality of life in patients having an ileal conduit, continent reservoir or orthotopic neobladder after cystectomy for bladder carcinoma. AB - BACKGROUND: To compare the QOL in patients with ileal or colon conduits (IC), continent urinary reservoir (CR) and ileal neobladder (NB), a retrospective study was conducted using a questionnaire sent by mail. METHODS: Seventy-nine patients with a mean age of 60 years were included in this study. A total of 36, 22 and 21 underwent IC, CR and NB, respectively and were alive at the time of this study. A structured questionnaire consisting of 97 questions that covered general condition and physical condition, reconstruction-related symptoms, psychological status, sexual life, social status and satisfaction with the treatment was employed. RESULTS: The IC group frequently complained of changes in bathing habits and loss of using public baths in comparison with the CR and the NB groups. High scores for loss of sexual desire were obtained in the IC, the CR and the NB groups, in this order. Because of the nearly physiological voiding, the NB group desired a voiding condition like pre-operative status as compared with the IC and the CR groups. However, for most of the questionnaire items no difference was seen among the IC, CR and NB groups concerning general condition, reconstruction-related symptoms, psychological status, sexual life, social status, satisfaction with the treatment and global satisfaction with life and health. CONCLUSIONS: There was little difference in the QOL score of the questionnaire and satisfaction among the IC, CR and NB groups. It was suggested that almost every patient accepted and adapted to the present status of general quality of life in each group. PMID- 10466452 TI - Diagnosis and treatment of pelvic lymph node metastasis in bladder control. AB - BACKGROUND AND METHODS: Bipedal lymphography and percutaneous fine needle aspiration biopsy (FNAB) of pelvic lymph node was done in 200 patients with bladder cancer. RESULTS: A diagnosis of metastasis to the pelvic lymph node was obtained by this method in 34 patients. Of these 34 patients, only 12 (35%) were positive or suspected of having pelvic lymph node metastasis by computed tomography. Sixteen patients (47%) had unequivocally positive or highly suspicious lymphogram and 18 patients (53%) had normal lymphogram. Seventy-eight cases, including eight FNAB-positive cases, were treated by radical cystectomy and regional lymph node dissection. Sensitivity, specificity, positive predictive value and negative predictive value of FNAB were 57, 100, 100 and 91%, respectively. Thirteen FNAB-positive M0 patients could be treated by multimodality therapy and evaluated after neoadjuvant therapy by repeated lymphography and FNAB. All were changed to N0 by FNAB after neoadjuvant therapy. In eight patients treated by total cystectomy and lymph node dissection, four had down staged to pN0 but four had one positive node. In five follow-up cases without surgery, one had recurrent pelvic lymph node metastasis 35 months later. Cause-specific survival of these patients was 76% at 5 years and 57% at 8 years. CONCLUSIONS: Pretreatment diagnosis of pelvic lymph node metastasis and multimodality therapy may improve the prognosis of regional lymph node-positive bladder cancer. Because it is difficult to diagnose pN1 using FNAB, radical operation should be performed even if neoadjuvant therapy is effective and down staging is obtained. PMID- 10466453 TI - External beam radiation monotherapy for prostate cancer. AB - BACKGROUND: To clarify the implications and limitations of external beam radiation monotherapy for localized prostate cancer, the long-term outcomes and prognostic factors were investigated. METHODS: Between 1976 and 1994, 91 patients with untreated prostate cancer were treated with external beam radiation therapy alone. Thirty-two were classified as T1b, eight were T2a, four were T2b and 47 were T3. Pelvic lymphadenectomy was carried out in 69 cases; 57 were staged as pN0, eight were pN1, four were pN2 and 22 were pNX. Linac X-rays were used in 55 cases, fast neutron in 15 and a combination of the two in 21. No other therapy was given until relapse and when relapse was evident endocrine therapy was started. RESULTS: The observation period ranged from 3 to 206 months with a median of 78 months. Local control rate and disease-free, cause-specific and overall survivals at 10 years were 74.0, 49.6, 74.2 and 39.2%, respectively. By univariate analysis, T category, pN category and histologic grade were significant prognostic indicators for disease-free survival. Multivariate analysis revealed that T category was an independent prognostic factor. In T2b and T3 diseases, pN0/1 patients demonstrated significantly better disease-free survival than pNX. CONCLUSIONS: A favorable long-term outcome was achieved by external beam radiation monotherapy in patients with minimally extended prostate cancer (T1b and T2a). For locally advanced disease (T2b and T3), staging pelvic lymphadenectomy would be useful for the selection of patients. PMID- 10466454 TI - Mesoblastic nephroma in adulthood: a case report. AB - BACKGROUND: Mesoblastic nephroma is an uncommon renal tumor of infancy and rarely occurs in adults. We report an adult case of mesoblastic nephroma. METHODS: A 50 year-old woman was found incidentally to have a right renal mass by abdominal ultrasonography. Computed tomography and magnetic resonance imaging revealed a heterogeneous tumor and angiography showed a mixture of hypervascularity and hypovascularity. Right radical nephrectomy was performed. RESULTS: The tumor was an encapsulated yellowish solid mass. Microscopically, the tumor was composed of spindle cell proliferation. Atypia and mitoses were not identified. Among the tumor cells, there were tubular arranged epithelial elements. CONCLUSION: The patient was free of recurrence 14 months postoperatively. Mesoblastic nephroma is classified as a benign tumor but recurrence and malignant formation of this tumor have been reported so regular postoperative follow up is required. PMID- 10466455 TI - Retroperitoneoscopic treatment of a retrocaval ureter. AB - BACKGROUND: Retroperitoneoscopic surgery was performed on a 41-year-old man with a retrocaval ureter. METHODS: The retrocaval segment of the ureter was retroperitoneoscopically separated and the ureter was reanastomosed using an automatic suture device. RESULTS/DISCUSSION: He returned to normal activities four days postoperatively. Postoperative excretory urography showed improvement of hydronephrosis and no residual stenosis. PMID- 10466456 TI - Neurogenic bladder dysfunction due to Behcet's disease. AB - BACKGROUND: Behcet's disease is a chronic multisystem condition of unknown origin. Genitourinary tract involvement with genital apthous ulcers, epididymitis, urethritis and recurrent cystitis is encountered commonly. Neurogenic bladder dysfunction due to neural involvement of the syndrome, has been reported extremely rarely. METHODS: A case of neurogenic bladder dysfunction due to Behcet's disease is presented. The patient was 17-year-old young male with a long history of the disease and neurological affection, but with neglected lower urinary tract symptoms and urinary incontinence. The patient was diagnosed urodynamically to have hyperreflexic detrusor. RESULTS/DISCUSSION: After failure of medical treatment (oxybutynin chloride) the patient submitted to augmentation clam-type sigmoidocystoplasty. Four years postoperatively, the patient is dry, on clean intermittent self catheterization twice daily and he is practically asymptomatic from lower urinary tract symptoms. PMID- 10466457 TI - How to assess endothelial function in human blood vessels. AB - This brief review discusses the ways, if and when available, to examine endothelium-dependent changes diameter in human blood vessels. It stresses the problems in ensuring proper matching between arteries (and veins) from different human sources. It briefly considers the evidence in vitro supporting the role of endothelium-derived nitric oxide, hyperpolarizing factor and contracting factors (including metabolites of arachidonic acid and endothelin). It emphasizes the difficulty in extrapolating observations obtained in isolated arteries (and veins) to the intact human circulation. The overall conclusion is that the interpretations derived from animal work apply to the human vasculature. PMID- 10466458 TI - Debating sympathetic overactivity as a hallmark of human obesity: a pro's position. PMID- 10466459 TI - Debating sympathetic overactivity as a hallmark of human obesity: an opposing position. PMID- 10466460 TI - Pulse pressure as a risk factor for cardiovascular events in the MRC Mild Hypertension Trial. AB - OBJECTIVES: The aim of this study was to determine whether pulse pressure is a risk factor for coronary artery disease using data from the MRC trial of treatment of mild hypertension, and whether the effect of anti-hypertensive drug therapy on pulse pressure may be a determinant of outcome in treated patients. METHODS: Logistic regression and Cox regression analyses were used to compare systolic and diastolic blood pressure, pulse pressure and mean blood pressure as predictors of coronary events and stroke in the MRC Mild Hypertension Trial. The effects of anti-hypertensive drug treatment with bendrofluazide and propranolol on pulse pressure were assessed using 1-year follow-up data. Event rates in the placebo-treated group and responses to anti-hypertensive treatment were measured in quartiles of age-adjusted entry pulse pressure. A 'four-corners' analysis was performed, with subjects divided into the upper and lower halves of the distributions of systolic and diastolic blood pressure at entry. RESULTS: Pulse pressure was a stronger predictor of coronary events than systolic, diastolic or mean blood pressure in males by logistic regression. Pulse pressure was similar to systolic pressure as a coronary event predictor on Cox regression. Stroke was predicted most strongly by mean blood pressure. Fatal and non-fatal coronary event rates increased progressively in ascending quartiles of age-adjusted pulse pressure, but there was also a strong correlation with systolic blood pressure. The values of partial logistic regression coefficients in models containing both systolic and diastolic blood pressure also supported a role for pulse pressure in predicting coronary events and for mean blood pressure in predicting stroke. Coronary risk, but not stroke, was inversely related to diastolic blood pressure in the four-corners analysis. In a Cox model, regressions of coronary event probability on systolic blood pressure at entry were significantly and inversely related to diastolic blood pressure categorized in quartiles. Bendrofluazide but not propranolol decreased pulse pressure significantly and was associated with a reduction in cardiovascular events overall, but no definite relationship between the effect of drugs on pulse pressure and specific responses to treatment was seen. CONCLUSION: Pulse pressure is a strong risk factor for coronary events in untreated hypertensive male subjects in the MRC Mild Hypertension Trial, whereas stroke is best predicted by mean blood pressure. Bendrofluazide and propranolol have different effects on pulse pressure which may be related to their relative efficacy in the treatment of hypertension, but this possibility requires further study in more suitable populations. PMID- 10466461 TI - End tidal CO2 is an independent determinant of systolic blood pressure in women. AB - OBJECTIVE: Recent studies have shown that high resting end tidal CO2 (PECO2) is a marker for blood pressure sensitivity to high sodium intake by normotensive humans. The present study investigated the association of resting PECO2 with resting blood pressure in participants in the Baltimore Longitudinal Study on Aging (BLSA). DESIGN: PECO2 of 314 healthy participants (153 men and 161 women, aged 21-79 years) was measured by a respiratory gas monitor during 25 min of seated rest, and blood pressure every 5 min by an automated oscillometric system. The independent associations of PECO2 and other variables with systolic (SBP) and diastolic (DBP) blood pressure were analysed via multiple regression. Sex differences in age-associated changes in PECO2 and in the role of pulmonary capacity in the PECO2 -blood pressure relationships were also studied. RESULTS: Resting PECO2 was an independent predictor of SBP in women (beta = 0.215; P<0.0015; overall r2 = 0.27; P<0.0001), and accounted for more than 10% of the variance in SBP in women over age 50 years. No such associations of PECO2 with SBP of men, or with DBP of men or women, were observed. PECO2 was lower in younger women (36.0+/-0.9 mm Hg) than in younger men (40.4+/-2.0), but higher in older women (37.1+/-0.5) than in older men (35.5+/-0.4). CONCLUSIONS: PECO2 is an independent determinant of resting SBP in women, especially those aged 50 years or more. The origin of individual differences in resting PECO2 and its possible role in the development of chronic hypertension remain to be clarified. PMID- 10466462 TI - Altered sodium pump alpha and gamma subunit gene expression in nephron segments from hypertensive rats. AB - OBJECTIVE: To determine the qualitative and quantitative expression of alpha and gamma sodium pump subunits in whole kidney and nephron segment RNA from Sprague Dawley rats, spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. DESIGN: A novel reverse transcription polymerase chain reaction technique was devised which provides accurate and precise measurement of the number of molecules of specific transcript abundance, a measurement of gene expression. This allows the quantitative comparison of multiple samples across multiple subjects and, since the estimates are accurate rather than relative, can also be used to make quantitative comparisons across expressed genes, such as isoforms and subunits of the heterotrimeric renal sodium pump. METHODS: We examined which catalytic isoforms were expressed and then quantified transcript abundance in whole kidney and convoluted and straight segments of the proximal tubule. RESULTS: Alpha 1 and gamma transcripts, but not alpha 2, alpha 3 or alpha 4 isoforms, were consistently observed in nephron segments. Levels of alpha 1 were lower in kidney RNA from 15-16-week-old SHR than in WKY rats of the same age (P = 0.001), but were not different between SHR and WKY in 4-5-week-old animals. No significant difference was observed in gamma subunit abundance in kidney RNA from 4-5-week-old animals; however, at 15-16 weeks, the expression in SHR was one third that in WKY rats (P = 0.003). In proximal convoluted tubules from 4-5-week old animals, the level of alpha 1 RNA expression was lower (P = 0.03) in SHR than in WKY rats. In addition, levels of alpha 1 in proximal straight tubule from the 4-5-week-old SHR were also lower than in WKY rats (P = 0.02). This difference was even greater in 15-16-week-old animals: in SHR, alpha 1 expression was less than 20% of the level of expression in WKY rats (P = 0.0003). Expression of the gamma subunit exhibited a similar pattern of downregulation in SHR. In RNA from proximal convoluted tubules and proximal straight tubules from both 4-5- and 15 16-week-old animals, expression of the gamma subunit was demonstrated to be significantly lower in SHR than in WKY rats. CONCLUSION: The results indicate a coordinate reduction in the abundance of sodium pump alpha and gamma subunits in the proximal tubules of SHR, which occurs early during the development of hypertension. PMID- 10466463 TI - Regulation of the genes for 11beta-hydroxysteroid dehydrogenase type 1 and type 2 in the kidney of the Dahl rat. AB - BACKGROUND: An isoenzyme of 11beta-hydroxysteroid dehydrogenase (11beta-HSD), 11beta-HSD-2 confers aldosterone specificity on the mineralocorticoid receptor (MR) and is found collocated in renal cortical collecting duct cells with the MR. To investigate whether the salt sensitivity of the Dahl salt-sensitive (S) rat is due to 11beta-HSD deficiency, we measured 11beta-HSD-1 and 11beta-HSD-2 mRNA levels in the kidneys of Dahl-S and Dahl salt-resistant (R) rats. In addition, we studied the effects of gender, age and dietary sodium on expression of mRNA for the two isoforms. S and R rats were placed on low- or high-sodium (HNa) diets and sacrificed after 33 and 115 days. Rat kidney RNA was isolated and 11beta-HSD-1 and 11beta-HSD-2 mRNA levels were measured on Northern filter hybridization using isoform-specific probes. RESULTS: No strain differences were observed in the mRNA expression of the two isoforms of 11beta-HSD under any of the experimental conditions. No gender or age differences were observed in 11beta-HSD-2 mRNA but HNa diet almost doubled 11beta-HSD-2 mRNA (P<0.0009). 11beta-HSD-1 mRNA levels were consistently higher, more than double, in male rats versus females rats (P<0.0001), and in the 115-day-old rats versus the 33-day-old rats (P<0.0001). Dietary sodium intake did not affect 11beta-HSD-1 mRNA levels. CONCLUSIONS: There is no difference in the expression of the two isoforms of 11beta-HSD in the kidneys of the S and R rats, which might explain the salt sensitivity and higher blood pressure of the S rat. Renal 11beta-HSD-1 mRNA levels are higher in male than in female rats, and in the older rats of both strains. In the kidney, the 11beta-HSD-2 gene is regulated by sodium status but is not affected by gender or age. PMID- 10466464 TI - G protein beta3 subunit 825T allele and its potential association with obesity in hypertensive individuals. AB - OBJECTIVE: The 825T allele of the G protein beta3 subunit is associated with hypertension in Caucasians. To generate a novel hypothesis regarding the underlying mechanisms, we examined for a potential association of the 825T allele with obesity in hypertension. PATIENTS AND METHODS: We genotyped 197 hypertensive individuals (104 men, 93 women; mean age 54 years) recruited from the general population in the Heidelberg (Germany) area. Data acquisition included age at first diagnosis of hypertension, body weight and height, actual treated blood pressure values, and history of stroke and/or myocardial infarction (cardiovascular events). RESULTS: The 825T allele was significantly (P = 0.02) associated with body mass index (BMI), mean values being 28.6+/-4.1, 27.0+/-3.1, and 26.1+/-3.8 kg/m2 for TT, TC, and CC respectively, which persisted after correction for sex and age. The 825T allele frequency was 23.8, 31.4 and 40.0% in individuals with normal weight (BMI <24.9 kg/m2), overweight (BMI 25.0-29.9 kg/m2), and obesity (BMI >29.9 kg/m2), respectively. Odds ratio for obesity versus normal weight was 3.9 [95% confidence interval (CI) 1.1-14.3; P = 0.03] for TT/CC and 1.8 (95% CI 0.7-4.6; P = 0.18) for TC/CC. BMI and age, but not genotype were significantly correlated with cardiovascular events as determined by logistic regression analysis. CONCLUSIONS: The findings suggest an association between obesity and the 825T allele, a genetic marker for enhanced G protein reactivity, in hypertensive individuals. PMID- 10466465 TI - Contribution of synthetic phenotype on the enhanced angiotensin II-generating system in vascular smooth muscle cells from spontaneously hypertensive rats. AB - OBJECTIVE: We have demonstrated that cultured vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR), but not from normotensive Wistar Kyoto (WKY) rats, produce angiotensin II (Ang II) in a homogeneous culture with increased levels of angiotensinogen, cathepsin D and angiotensin converting enzyme (ACE) at early passages. In the current study, we investigated how changes in the cell phenotype affect the Ang II-generating system and the growth of VSMC from SHR. DESIGN AND METHODS: We evaluated basal DNA synthesis by [3H]thymidine incorporation, immunofluorescence of alpha-smooth muscle (SM) actin, mRNA expression of phenotype markers such as SM22alpha appeared by contractile phenotype, Ang II-generating system components and growth factors by reverse transcription and polymerase chain reaction analysis, and Ang II levels by radioimmunoassay in quiescent VSMC from WKY/Izumo rats and SHR/Izumo at passages 4, 8 and 12. RESULTS: Basal DNA synthesis in VSMC from WKY rats increased with increasing passage number, whereas in cells from SHR it was markedly higher at early passages and was not affected by the passages. At early passage numbers, immunofluorescence of alpha-SM actin was stronger in VSMC from WKY rats than in cells from SHR, but decreased after several passages. Expression of SM22alpha mRNA was higher in VSMC from WKY rats than in cells from SHR at early passages, and decreased after several passages in cells from both rat strains. Expression of matrix Gla mRNA was higher in VSMC from SHR than in cells from WKY rats at early passage, and increased after several passages in cells from both rat strains. Ang II was not detected at early passages but increased in VSMC from WKY rats with increasing passage, whereas it was detected in VSMC from SHR at early passages and did not change with the passages. Expression of angiotensinogen mRNA was higher in VSMC from SHR than in cells from WKY rats, and was not affected by the passages. Expressions of cathepsin D and ACE mRNA were higher in VSMC from SHR than in cells from WKY rats at early passage, and were increased by the passages in VSMC from WKY rats. Expressions of transforming growth factor-beta1, platelet-derived growth factor A-chain, and basic fibroblast growth factor mRNA were significantly higher in VSMC from SHR than in cells from WKY rats, and were increased by the passages. CONCLUSION: These data indicate that early in culture VSMC from SHR have the synthetic phenotype, whereas VSMC from WKY rats have the contractile phenotype which then changes to the synthetic phenotype after increased passage numbers, with increased expression of cathepsin D and ACE, which produce Ang II, and increased expression of Ang II-related growth factors, which induce the exaggerated growth observed in VSMC from SHR. PMID- 10466466 TI - Pressure but not angiotensin II-induced increases in wall mass or tone influences static and dynamic aortic mechanics. AB - OBJECTIVE: To distinguish between static (due to slow changes in pressure) and dynamic (due to pressure pulsatility) components of aortic compliance over a large pressure range in vivo and to examine the effects of increased vascular mass and smooth muscle tone on these components. METHODS: Using ultrasound wall tracking, aortic lumen area-pressure curves were generated in anaesthetized rats over a broad range of pressures by altering blood volume. The compliance coefficient calculated at each mean pressure was considered the dynamic compliance at that pressure; the slope of the diastolic lumen area-pressure curve represents static compliance. Experiments were performed in control rats and rats treated with angiotensin II (ANG II) acutely (500 ng/kg per min intravenously) to modify vascular tone or chronically (250 ng/kg per min subcutaneously for 2 weeks) to modify vascular mass. RESULTS: The dynamic compliance-pressure curve approximated a parabola. Maximal dynamic compliance (0.272+/-0.026 mm2/kPa in control rats) was achieved at near-normotensive pressure (+/-105 mm Hg). The diastolic lumen area-pressure curve showed an exponential relationship within a physiological range (30-130 mm Hg). ANG II-induced increases in aortic wall mass or smooth muscle tone did not modify the relationship between static or dynamic compliance and pressure. CONCLUSIONS: These findings demonstrate that static and dynamic mechanics of the rat thoracic aorta depend differently on blood pressure. Static compliance increases slightly with pressure in a physiological range, while dynamic compliance is auto-regulated around normotensive pressures. Neither static nor dynamic compliance of the rat thoracic aorta are influenced by ANG II induced increases in aortic wall mass or smooth muscle tone. PMID- 10466467 TI - Sympathetic tone restrains arterial distensibility of healthy and atherosclerotic subjects. AB - BACKGROUND: Sympathetic activation induced by cold pressor test or cigarette smoking is accompanied by a marked reduction of radial artery distensibility. It is not known, however, whether arterial distensibility is under tonic sympathetic restraint, or whether this restraint involves arteries greater than the radial one in both normal and pathological conditions. METHODS: We studied the distensibility of radial artery by continuous ultrasonographic assessment of the changes in arterial diameter over the diasto-systolic pressure range (finger pressure measurement) in eight patients with a Dupuytren disease before and 20 min after ipsilateral brachial plexus anaesthesia. We also studied ultrasonographic distensibility of femoral artery in seven subjects before and 20 min after ipsilateral subarachnoid anaesthesia, performed before arthroscopic surgery, and in five patients with claudicatio intermittens before and 1 month after ipsilateral removal of the lumbar sympathectomy chain. In all three conditions, the contralateral artery served as control. RESULTS: The three interventions did not cause any significant alteration in blood pressure and heart rate. Radial artery distensibility was markedly increased by ipsilateral anaesthesia of the brachial plexus (+36%, P<0.01). This was the case also for femoral artery distensibility both following ipsilateral subarachnoid anaesthesia in healthy subjects (+47%, P<0.05) or ipsilateral sympathetic gangliectomy in patients with peripheral artery disease (+26%, P<0.05). In all three instances, the distensibility of the contralateral artery remained unaffected. CONCLUSIONS: These data indicate that the sympathetic nervous system exerts a marked tonic restraint of arterial distensibility. This restraint involves medium-size and large muscular arteries and can also be seen in subjects with peripheral artery disease. This stiffening influence may increase the traumatic effect of intravascular pressure on the vessel wall and favour atherosclerosis. PMID- 10466468 TI - Neural mechanisms in human obesity-related hypertension. AB - OBJECTIVE: Two hypotheses concerning mechanisms of weight gain and of blood pressure elevation in obesity were tested. The first hypothesis is that in human obesity sympathetic nervous system underactivity is present, as a metabolic basis for the obesity. The second hypothesis, attributable to Landsberg, is that sympathetic nervous activation occurs with chronic overeating, elevating blood pressure. These are not mutually exclusive hypotheses, since obesity is a heterogeneous disorder. DESIGN AND METHODS: Whole body and regional sympathetic nervous system activity, in the kidneys and heart, was measured at rest using noradrenaline isotope dilution methodology in a total of 86 research voluteers in four different subject groups, in lean and in obese people who either did, or did not, have high blood pressure. RESULTS: In the lean hypertensive patients, noradrenaline spillover for the whole body, and from the heart and kidneys was substantially higher than in the healthy lean volunteers. In normotensive obesity, the whole body noradrenaline spillover rate was normal, mean renal noradrenaline spillover was elevated (twice normal), and cardiac noradrenaline spillover reduced by approximately 50%. In obesity-related hypertension, there was elevation of renal noradrenaline spillover, comparable to that present in normotensive obese individuals but not accompanied by suppression of cardiac noradrenaline spillover, which was more than double that of normotensive obese individuals (P<0.05), and 25% higher than in healthy volunteers. There was a parallel elevation of heart rate in hypertensive obese individuals. CONCLUSIONS: The sympathetic underactivity hypothesis of obesity causation now looks untenable, as based on measures of noradrenaline spillover, sympathetic nervous system activity was normal for the whole body and increased for the kidneys; the low sympathetic activity in the heart would have only a trifling impact on total energy balance. The increase in renal sympathetic activity in obesity may possibly be a necessary cause for the development of hypertension in obese individuals, although clearly not a sufficient cause, being present in both normotensive and hypertensive obese individuals. The discriminating feature of obesity-related hypertension was an absence of the suppression of the cardiac sympathetic outflow seen in normotensive obese individuals. Sympathetic nervous changes in obesity-related hypertension conformed rather closely to those expected from the Landsberg hypothesis. PMID- 10466469 TI - Changes in short-term variability of blood pressure and heart rate during the development of obesity-associated hypertension in high-fat fed dogs. AB - OBJECTIVE: To investigate the nature and time course of autonomic nervous system changes elicited by a 21-week ad libitum high-fat diet (HFD) in dogs. RESULTS: The HFD increased body weight (+22.0+/-2.8% at week 21) with an abdominal circumference gain significantly more elevated than the thoracic one. The increases in insulin and free fatty acid plasma levels were correlated with body weight changes. Systolic and diastolic blood pressures and heart rate significantly increased (+23+/-6, +28+/-5 and 19+/-9% respectively). Arterial hypertension was characterized by an increase in cardiac output (+22.3+/-7.7%), in left ventricular mass (+18.1+/-5.0% at week 21) and a decrease in spontaneous baroreflex efficiency (-55+/-6%). The time course of autonomic changes (using spectral analysis of systolic blood pressure and heart rate) showed the existence of time-dependent modifications, which were linked with food intake. The initial rise in arterial blood pressure during body weight increment (observed between the 1st and 8th week of HFD) was associated with a transient increase in the low frequency band of systolic blood pressure variability and noradrenaline plasma levels associated with a long-lasting decrease in the high frequency band of heart rate variability. Early changes in short-term variability were significantly correlated with free fatty acid plasma levels. In contrast, the steady-state of obesity-related hypertension was associated with a decreased high frequency band of heart rate variability, without significant changes in noradrenaline plasma levels. CONCLUSIONS: This study shows that the HFD induces abdominal obesity, hyperinsulinaemia and arterial hypertension, with a left ventricular hypertrophy associated with a biphasic changes in autonomic activity: an early and long-lasting decrease in parasympathetic nervous system activity and an early but transient increase in sympathetic activity. The present data suggest that autonomic nervous system changes are dependent on the time course of obesity development. PMID- 10466470 TI - Haemorrhage increases the pressor effect of angiotensin-(1-7) but not of angiotensin II at the rat rostral ventrolateral medulla. AB - OBJECTIVE: To evaluate the effects of angiotensins acting at the rostral ventrolateral medulla (RVLM) on the cardiovascular adjustments following haemorrhage. DESIGN: Changes in mean arterial pressure (MAP) and heart rate (HR) produced by micro-injections of angiotensin II (Ang II) and angiotensin (Ang)-(1 7) and different angiotensin antagonists into the RVLM of anaesthetized rats submitted to haemorrhage, were determined. METHODS: Experiments were performed in 79 urethane-anaesthetized male Wistar rats. Ang-(1-7) (2.5 and 25 pmol), Ang II (25 pmol), [Sar1,Thr8]-Ang II (non-selective angiotensin antagonist, 0.2 nmol), A 779 (Ang-(1-7) antagonist, 0.1 nmol), losartan (AT1 Ang II receptor antagonist, 0.2 nmol) or vehicle (200 nl) were bilaterally micro-injected into the RVLM under basal conditions or 30 min after blood withdrawal (0.6 ml/100 g bodyweight). In additional groups, [Sar1,Thr8]-Ang II, A-779, losartan or vehicle were micro injected into the RVLM 10 min before bleeding to uncover a possible role of endogenous peptides during haemorrhage. RESULTS: The pressor effect produced by Ang II micro-injection was not altered by haemorrhage. Conversely, haemorrhage significantly increased the magnitude and duration of the pressor effect of Ang (1-7) at the RVLM. The fall in MAP induced by haemorrhage was similar after micro injection of vehicle or A-779. However, micro-injection of [Sar1,Thr8]-Ang II significantly reduced the fall in MAP after haemorrhage. A similar finding was obtained with micro-injection of losartan. In addition, while RVLM micro injection of [Sar1,Thr8]-Ang II or losartan 30 min after blood withdrawn produced MAP changes that were similar to that observed in control animals, micro injection of A-779 did not significantly alter baseline MAP. CONCLUSIONS: These results suggest that changes in the RVLM reactivity to Ang-(1-7) but not Ang II may contribute to the haemodynamic adjustments triggered by acute reductions in blood volume. The data obtained with [Sar1,Thr8]-Ang II and losartan suggest a primary inhibitory role for endogenous Ang II at the RVLM during haemorrhage. PMID- 10466471 TI - Effect of insulin resistance on left ventricular hypertrophy and dysfunction in essential hypertension. AB - BACKGROUND: In hypertensive patients, the relationships between glucose tolerance and left ventricular hypertrophy (LVH) and left ventricular diastolic function (LVDF) have been described in several reports. OBJECTIVE: In this study, we examined the relationships between insulin resistance and LVH and LVDF in hypertensive patients from the therapeutic perspective. METHODS AND RESULTS: The study participants were essential hypertensive patients with impaired glucose tolerance (IGT-HT, n = 26), hypertensive patients with normal glucose tolerance (NGT-HT, n = 39), and normotensive control individuals (n = 18). Insulin resistance was evaluated by the insulin suppression test by use of the steady state plasma glucose (SSPG) level. Left ventricular mass index (LVMI) and LVDF, which was determined by the E:A ratio, were estimated by echocardiography. Temocapril, an angiotensin-converting enzyme inhibitor, was administered in an open, non-randomized manner with a mean dose of 2.8+/-0.2 mg/ day, and the mean administration period was 18 weeks. The systolic and diastolic blood pressure, the LVMI, and the SSPG level were significantly higher in the hypertensive patients than in the control individuals. The mean systolic and diastolic blood pressures were significantly decreased by treatment with Temocapril. Before treatment, stepwise regression analysis showed that SSPG is an independent predictor for LVMI and LVDF. After treatment, the changes in LVMI (D-LVMI; %) ( 15.1+/-1.5), the changes in LVDF (D-E:A; %) (-38.2+/-4.1), and the changes in insulin resistance (D-SSPG; %) (-13.7+/-1.7) were significantly higher in the IGT HT group than in the NGT-HT group (-11.4+/-1.1, -18.1+/-1.7, -9.4+/-1.4, respectively), and the D-SSPG was an independent predictor for D-LVMI and D-E :A. CONCLUSIONS: The results of this study indicate that insulin resistance is an important factor affecting LVH and LVDF. PMID- 10466472 TI - Relationship between insulin resistance and cardiac sympathetic nervous function in essential hypertension. AB - BACKGROUND: It has been suggested that hyperinsulinemia and insulin resistance participate in the pathogenesis of hypertension, in part by activating sympathetic activity. OBJECTIVE: We aimed to examine the relationship between insulin resistance and cardiac sympathetic nervous function in patients with essential hypertension using 123I-metaiodobenzylguanidine (MIBG) cardiac scintigraphy. METHODS AND RESULTS: Twenty-eight patients (18 men) with essential hypertension and 11 (seven men) control individuals with a mean age of 55.8+/-3.3 years were recruited. Patients with diabetes mellitus, congestive heart failure or coronary artery disease were excluded from this study. To evaluate insulin resistance, we used steady-state plasma glucose (SSPG; mg/dl) levels measured by the SSPG method. To evaluate cardiac sympathetic nervous function, we calculated the heart-to-mediastinum ratio from the delayed MIBG image (H:M-D) and the mean washout rate (WOR, %). There were significant differences (P<0.01) in SSPG, H:M-D and WOR between the essential hypertension and control individual groups (125 versus 103 mg/dl, 2.2 versus 2.4, and 32 versus 23%, respectively). Stepwise regression analysis showed that SSPG and plasma norepinephrine level are independent predictors for the cardiac sympathetic nervous function obtained from MIBG scintigraphy. CONCLUSIONS: These findings indicate that insulin resistance is significantly related to activation of the cardiac sympathetic nervous function associated with left ventricular hypertrophy in patients with essential hypertension. PMID- 10466473 TI - Elevated fasting insulin predicts incident hypertension: the ARIC study. Atherosclerosis Risk in Communities Study Investigators. AB - OBJECTIVE: The prospective association of insulin and hypertension has been under debate in the context of the development of the insulin resistance or multiple metabolic syndrome. We examined the predictive associations of fasting serum insulin with incident hypertension occurring alone or as part of the multiple metabolic syndrome. DESIGN: Analyses were restricted to 5221 middle-aged participants of the Atherosclerosis Risk in Communities Study cohort who were free of component disorders of the multiple metabolic syndrome (hypertension; diabetes; high triglycerides and/or low HDL cholesterol (dyslipidaemias)) at baseline. OUTCOME: A total of 1018 individuals developed hypertension, 801 in the absence of components of the metabolic syndrome and 217 in combination with diabetes or dyslipidaemias, between 1987 and 1993. RESULTS: Elevated fasting insulin (top quartile versus lowest quartile) was associated with overall incident hypertension in European Americans [hazard rate ratio (HRR) 2.0, 95% confidence interval (CI) 1.7-2.4] but the results were inconclusive in African Americans (HRR 1.3, 95% CI 0.9-1.8) after adjustment for age, gender and study centre. Among European Americans, body mass index and abdominal girth only partly explained the observed association. Elevated fasting insulin was more strongly predictive of hypertension occurring as a component of the multiple metabolic syndrome (HRR 2.4, 95% CI 1.5-3.9) than of hypertension occurring alone (HRR 1.3, 95% CI 1.0-1.7) adjusting statistically for age, gender, study centre, body mass index and abdominal girth. CONCLUSIONS: The results are consistent with the concept of an aetiological heterogeneity for hypertension and may explain previously reported inconsistent findings on the association of insulin with incident hypertension. PMID- 10466474 TI - Circulating bufodienolide and cardenolide sodium pump inhibitors in preeclampsia. AB - OBJECTIVE: To determine plasma levels of the endogenous bufodienolide Na+/K+ ATPase inhibitor, marinobufagenin-like factor (MBG), in normotensive pregnancy and in preeclampsia, to compare changes of MBG with that of ouabain-like compound (OLC), and to characterize the purified MBG immunoreactive factor from preeclamptic plasma. DESIGN AND METHODS: Consecutive sample study. The levels of MBG and OLC compounds were measured in extracted plasma by solid phase fluoroimmunoassays. MBG and ouabain immunoreactive materials were partially purified from preeclamptic plasma via reverse-phase high-performance liquid chromatography (HPLC) and studied for their ability to cross react with MBG and ouabain antibodies, and to inhibit the Na+/K+ ATPase from human mesenteric arteries. Vasoconstrictor effect of authentic MBG was studied in isolated rings of human umbilical arteries. RESULTS: In 11 nonpregnant control individuals, plasma concentrations of MBG and OLC were 0.190+/-0.04 nmol/l and 0.297+/-0.037 nmol/l, respectively. In the third trimester of noncomplicated pregnancy (n = 6), plasma MBG increased (0.625+/-0.067 nmol/l, P<0.05), and OLC did not (0.32+/-0.07 nmol/l). In 15 patients with preeclampsia, plasma levels of both MBG and OLC increased dramatically (2.63+/-0.10 nmol/l and 0.697+/-0.16 nmol/l, respectively, P<0.01 versus both control groups). When fractionated by reverse phase HPLC, OLC was eluted by 18% acetonitrile, and MBG by 48% acetonitrile. Serially diluted samples of MBG and OLC immunoreactive materials from HPLC fractions reacted with MBG and ouabain antibody in solid phase immunoassay in a concentration dependent fashion. Authentic MBG caused contractile responses of isolated rings of human mesenteric arteries in a concentration-dependent manner. Similarly to the authentic MBG, HPLC purified MBG immunoreactive material from preeclamptic plasma inhibited Na+/K+ ATPase purified from human mesenteric artery. CONCLUSIONS: Our observations demonstrate the coexistence of two endogenous cardiotonic steroids in preeclamptic plasma, a more polar OLC and a less polar MBG-like compound. Substantial increases in plasma OLC and MBG immunoreactivity in preeclampsia, along with the vasoconstrictor properties of authentic MBG and Na+,K+ ATPase inhibitory activity of human MBG immunoreactive factor, suggest, that in preeclampsia, plasma concentrations of MBG are enough to substantially inhibit the sodium pump in cardiovascular tissues, and are in accordance with the views attributing endogenous digitalis-like factors a pathogenic role in the preeclamptic hypertension. PMID- 10466475 TI - Short-term metabolic effects of isradipine and metoprolol in pre-eclampsia. AB - OBJECTIVE: To study the effects of isradipine or metoprolol on insulin sensitivity and lipid profiles as well as on blood pressure and umbilical vascular resistance in pre-eclamptic women in the third trimester of pregnancy. DESIGN: A single-centre, prospective, randomized, double-blind, double-dummy and parallel-group study. SETTING: Helsinki University Central Hospital, a tertiary referral centre. PATIENTS: Twenty-four previously healthy pregnant women with normal findings in an oral glucose-tolerance test who were hospitalized for preeclampsia, of whom 17 completed the study. INTERVENTIONS: Between 29 and 39 weeks of gestation, measurements were made of insulin sensitivity (the minimal model), magnitude of proteinuria, and the fasting levels of serum uric acid, lipids and lipoproteins. Subsequently, treatment with isradipine 2.5 mg (n = 9) or metoprolol 50 mg (n = 8) twice daily was started, and these women were reinvestigated 5-7 days later. Blood pressure was recorded during 24 h by automated ambulatory blood pressure measurement. Umbilical artery resistance index was measured by Doppler ultrasound. MAIN OUTCOME MEASURES: Insulin sensitivity, uric acid, degree of proteinuria, lipids and lipoproteins, blood pressure, umbilical artery resistance index. sensitivity, degree of proteinuria, blood pressure, or the umbilical artery resistance index. Serum uric acid increased in both groups (P<0.05). High-density lipoprotein2 cholesterol increased 15.6% in the isradipine group (P<0.05), but no significant changes appeared in other lipids and lipoproteins in either group. CONCLUSIONS: In this study, short-term antihypertensive treatment with isradipine or metoprolol in preeclampsia had no detrimental effect on serum lipid and lipoprotein levels or insulin sensitivity. PMID- 10466476 TI - Evaluation of the potential interaction between NaCl and prostaglandin inhibition in elderly individuals with isolated systolic hypertension. AB - OBJECTIVE: To evaluate whether prostaglandin inhibition with the non-steroidal anti-inflammatory drug (NSAID), indomethacin (I) interacts synergistically with different doses of salt (NaCl) in elevating systolic blood pressure (SBP). DESIGN AND METHODS: This randomized, placebo-controlled, double-blind, crossover study examined the interaction between NaCl and the prostaglandin inhibitor, I in 31 healthy elderly individuals with a mean age (+/- SD) of 68.7+/-5.7 years (range 61-85 years). Participants aged more than 60 years on a 140 mmol/day NaCl dose for 6 weeks were chosen with normal blood pressure [24-h SBP <148 mm Hg, diastolic blood pressure (DBP) <85 mm Hg on the Takeda Ambulatory Blood Pressure Monitor (TABPM); n = 15] and isolated systolic hypertension (ISH), [24-h SBP >148 mm Hg, 24-h DBP <85 mm Hg on TABPM; n = 16]. Exclusion criteria included uncontrolled hypertension (SBP >220 mm Hg and/or DBP >110 mm Hg), cardiac disease, creatinine clearance <60 ml/min, dementia and recent cerebrovascular accident or secondary hypertension. A 2x2 Latin square design was structured using four treatment groups [low salt (NaCl = 90 mmol/day) + I placebo, high salt (NaCl = 240 mmol/day) + I placebo, low salt + I (25 mg three times daily) and high salt + I] for 2 weeks each, balanced and interspersed with 2 week washout periods to minimize carryover effects. Twenty-four hour SBP, DBP and heart rate were measured and summarized using a moving interval averaging technique. The mean change in 24-h SBP, DBP, heart rate, urinary Na+, K+, protein and creatinine, creatinine clearance and serum electrolytes were compared across treatments in the total cohort and in ISH and control groups separately using ANCOVA (SAS). RESULTS: In the total cohort, compared with low NaCl, chronic high NaCl increased mean SBP (5.76 mm Hg; P = 0.0002) and DBP (3.36 mm Hg; P = 0.002). Indomethacin significantly increased mean SBP (2.66 mm Hg, P = 0.015) but not DBP (0.31 mm Hg, P = 0.419). High salt and I were additive (SBPT, DBPT) but there was no interaction (P = 0.795 and P = 0.739, respectively). Additionally, chronic high NaCl increased serum Na (P = 0.0001) and 24-h urinary Na (P = 0.0001) as expected. Indomethacin significantly decreased mean heart rate (P = 0.018). The effects of NaCl and I on SBP, DBP and heart rate were not modified by age, alcohol intake, serum K+, body mass index or treatment order. In the ISH group, NaCl dose significantly elevated SBP (9.87 mm Hg; P = 0.0001) and DBP (5.26 mm Hg, P = 0.006) but did not significantly alter blood pressure in the normotensive group. Indomethacin significantly elevated SBP (P = 0.03) in normotensive individuals but had no effect on blood pressure in the ISH group. CONCLUSIONS: Chronic high salt diet elevated blood pressure more than I in the total cohort of elderly individuals. No interaction was demonstrated and their effects were additive. In the ISH group, chronic high salt diet significantly increased SBP and DBP while I failed to alter blood pressure. In the normotensive group, I, but not salt, elevated SBP. Patients with ISH are sensitive to the pressor effect of NaCl but resistant to the pressor effect of prostaglandin inhibition in contrast to elderly normotensive control individuals where the reverse was found. PMID- 10466477 TI - Prothrombotic factors, endothelial function and left ventricular hypertrophy in isolated systolic hypertension compared with systolic-diastolic hypertension. AB - BACKGROUND: Individuals with systolic-diastolic hypertension (SDH, systolic blood pressure (SBP) >160 mm Hg and diastolic blood pressure (DBP) >90 mm Hg) are at increased risk of thrombotic complications, such as stroke and heart attacks, which may be related to a hypercoagulable state. Individuals with only isolated systolic hypertension (ISH; i.e. SBP >160 mm Hg but DBP <90 mm Hg) are also at significant cardiovascular risk. We hypothesized that patients with ISH would exhibit a prothrombotic state similar to that seen in SDH. A secondary hypothesis was that individuals with ISH had similar echocardiographic parameters to those seen in SDH. METHODS: We measured indices of haemorheology, endothelial dysfunction, thrombogenesis and platelet activation in 23 individuals with ISH (mean blood pressure 193/82 mm Hg), who were compared with 51 matched patients with SDH (mean blood pressure 198/112 mm Hg) and 34 age- and sex- matched normotensive healthy control individuals (mean blood pressure 130/78 mm Hg). Echocardiographic parameters in patients with ISH were compared to those from patients with SDH. RESULTS: Mean plasma viscosity (an index of blood rheology, ANOVA, P = 0.001), von Willebrand factor (an index of endothelial damage, P = 0.013), plasminogen activator inhibitor-1 and lipoprotein (a) (both markers of thrombogenesis; Kruskal-Wallis test, both P<0.001) were all significantly raised in ISH and SDH relative to controls. Individuals with SDH also had high mean plasma fibrinogen (P = 0.018) and haematocrit (P = 0.010) levels compared with control individuals. There were no significant differences in levels of fibrin D dimer or the platelet activation marker soluble P-selectin in the hypertensive patients (i.e. ISH and SDH) compared with control individuals. Patients with ISH had similar M-mode and Doppler echocardiographic parameters compared to patients with SDH. CONCLUSIONS: We conclude that individuals with ISH have abnormalities in plasma prothrombotic factors and markers of endothelial dysfunction, and echocardiographic parameters, broadly similar to that seen in SDH. This is consistent with the increased risk of thrombotic events (strokes and heart attacks) in patients with ISH. PMID- 10466478 TI - Pharmacologic agents on cardiovascular mass, coronary dynamics and collagen in aged spontaneously hypertensive rats. AB - OBJECTIVE: To determine whether antihypertensive treatment could alter hypertension and age-related progressive impairment of coronary hemodynamics and cardiac fibrosis in aged spontaneously hypertensive rats (SHR). DESIGN: Old SHR were given their respective therapy for 3 months. To differentiate between hypertension and age-related changes, a comparison was made between left and right ventricular indices since the right ventricle was not exposed to pressure overload. METHODS: Male, 65-week-old spontaneously SHR were divided into three groups and were given either vehicle, felodipine (30 mg/kg per day) or enalapril (30 mg/kg per day). After 12 weeks of the respective treatments, systemic and coronary hemodynamics (radionuclide-labelled microspheres), right and left ventricular and aortic mass indices, and right and left ventricular hydroxyproline concentrations (an estimate of collagen) were determined. RESULTS: Arterial pressure and total peripheral resistance were reduced to the same extent in SHRs treated with either felodipine or enalapril; however, compared to the control rats, enalapril was more effective in reducing left ventricular and aortic mass indices. Both agents also improved coronary hemodynamics of both ventricles in aged SHR but enalapril was more effective as indicated by a greater increase in coronary flow reserve and a greater decrease in minimal coronary vascular resistance. Furthermore, enalapril but not felodipine reduced left ventricular hydroxyproline concentration; and right ventricular hydroxyproline concentration increased with felodipine but remained unchanged with enalapril. CONCLUSIONS: Both enalapril and felodipine ameliorated adverse cardiovascular effects of hypertension in the aged SHRs within 12 weeks, as demonstrated by reduced arterial pressure, diminished left ventricular mass, and improved coronary hemodynamics. Enalapril also decreased aortic mass and left ventricular collagen concentration and appeared to be more effective in improving coronary hemodynamics than felodipine, possibly as a result, in part, of reduced myocardial fibrosis. PMID- 10466479 TI - The effect of losartan and captopril on glomerular basement membrane anionic charge in a diabetic rat model. AB - OBJECTIVE: Although angiotensin-converting enzyme inhibitors are known to reduce albuminuria by preserving glomerular basement membrane anionic content, the effects of angiotensin II receptor blockage are currently not known. The aim of this study was to evaluate the effects of captopril and losartan on glomerular basement membrane anionic charges in a diabetic rat model. DESIGN: After diabetes induction with streptozotocin, female Wistar rats were divided into three groups: group A, losartan 10 mg/kg by gavage (n = 8); group B, captopril 50 mg/l in drinking water (n = 6); group C, diabetic control rats (n = 8) given only tap water. Group D (eight rats) served as non-diabetic controls. At the end of 8 weeks, erythrocyte membrane charge, serum sialic acid, urinary glycosaminoglycan and albumin were measured and kidney specimens stained with Alcian blue in order to assess basement membrane glycosaminoglycans. RESULTS: Red blood cell anionic charges (ng Alcian blue/ 10(6) red blood cells) were 371.5+/-9.9 for group A, 443.5+/-7.1 for group B, 400.1+/-14.7 for group C, 468.7+/-4 for group D (AD, P<0.01; A>B P<0.01). Albuminuria (microg/day) was 778+/-221 for group A, 719+/ 314 for group B, 1724+/-945 for group C, 393+/-263 for group D (A, B or = 3 beats (72%) and/or symptoms (68%). Sustained VT was induced in 25% of the studies, and nonsustained VT in 12%. Atrial flutter or other supraventricular tachycardia was documented in 32% and bradyarrhythmias in 26%. By univariate analysis, mortality was increased in patients with positive VSTIM versus negative VSTIM (18% vs 7%, P = 0.04). Using multivariate analysis, positive VSTIM was associated with a sixfold increased risk of decreased survival and a threefold increased risk of serious arrhythmic events, allowing up to 87% sensitivity in predicting mortality. However, 7 (33%) of 21 patients with documented clinical VT had false negative studies. CONCLUSION: VSTIM in a large, selected group of congenital heart disease patients identified a subgroup with significantly increased mortality and sudden arrhythmic events. Failure to induce VT was a favorable prognostic sign, but the frequency of false-negative studies was high. Frequent supraventricular tachycardia further complicated risk stratification. Although VSTIM appears to be a reasonable tool for evaluation of this population, a larger, multicenter trial is recommended to clarify its utility. PMID- 10466483 TI - Risk assessment of patients having congenital heart disease using electrophysiologic testing: finally, opening the right door...or, an impossible chore? PMID- 10466484 TI - Sudden death in recipients of transvenous implantable cardioverter defibrillator systems: terminal events, predictors, and potential mechanisms. AB - INTRODUCTION: Transvenous implantable cardioverter defibrillator (ICD) systems are very effective in preventing sudden death; however, little is known about terminal events and potential causes and mechanisms of sudden death in recipients of these devices. METHODS AND RESULTS: We analyzed 74 cases of sudden death among patients enrolled in several clinical investigations of transvenous ICD systems. Eighty-one percent were men (mean age 68+/-10 years), 86% had coronary artery disease, mean left ventricular ejection fraction was 0.27+/-0.11, and two thirds presented with sustained ventricular tachycardia. The final event was witnessed in 65 patients (81%). Based on reported ICD shocks, documented rhythm, and/or postmortem device data, sudden death was deemed tachyarrhythmic in 49 cases (66%), nontachyarrhythmic in 12 (16%), and indeterminate in the remaining 13 (18%). Multivariate analysis of several clinical and nonclinical factors found advanced age (> 65 years, P = 0.03, odds ratio [OR] 1.75, 95 % confidence interval [CI] 1.05 to 2.92), reduced left ventricular ejection fraction (< 0.35, P < 0.01, OR 3.51, CI 1.66 to 7.40), and having antibradycardia pacing ICDs (P = 0.02, OR 5.26, CI 1.37 to 20.0) to be independent predictors of sudden death. One or more predisposing factors and/or potential causes of sudden death were identified in 21 patients (28%). CONCLUSION: In this select group of transvenous ICD recipients, (1) sudden death was associated with ventricular tachycardia/ventricular fibrillation in at least two thirds of cases, (2) nearly one third of patients had one or more factors, some device related, that could have been associated with sudden death, and (3) death ensued despite appropriate ICD therapies and, in many cases, external resuscitation, suggesting acute adverse events as common terminal factors. PMID- 10466485 TI - Classification of death in clinical trials: precision versus accuracy. PMID- 10466486 TI - Characterization of sinoatrial parasympathetic innervation in humans. AB - INTRODUCTION: The response to sinoatrial parasympathetic nerve stimulation (shortened atrial refractoriness) was used to determine the atrial distribution of these nerve fibers in humans. We hypothesized that, in humans, parasympathetic nerves that innervate the sinoatrial node also innervate the right atrium and that the greatest density of innervation is near the sinoatrial nodal fat pad. METHODS AND RESULTS: Temporary epicardial wire electrodes were sutured in pairs in the sinoatrial nodal fat pad, high right atrium, and right ventricle by direct visualization during coronary artery bypass surgery in nine patients. Appropriate electrode placement was confirmed by electrically stimulating the fat pad in the operating room to prolong sinus cycle length by 50%. Experiments were performed in the electrophysiology laboratory 1 to 5 days after surgery. Programmed atrial stimulation was performed via an endocardial electrode catheter advanced to the right atrium. The catheter tip electrode was moved in 1-cm concentric zones around the epicardial wires by fluoroscopic guidance. Atrial refractoriness was determined in the presence and absence of sinoatrial parasympathetic nerve stimulation at each catheter site. In 8 of 9 patients, parasympathetic nerve stimulation reproducibly prolonged sinus cycle length by 50%. There was no effect on AV nodal conduction (no prolongation of PR interval) and no change in AV nodal refractoriness. Atrial effective refractory periods reproducibly shortened in response to parasympathetic nerve stimulation in 1-cm zones up to 3 cm surrounding the fat pad, by a mean (+/- SEM) of 26.6+/-4.3 msec (zone 1), 11.4+/ 1.8 msec (zone 2), and 10.0+/-2.5 msec (zone 3), respectively (P = 0.0001). At distances > 3 cm from the fat pad, the effective refractory period did not shorten. CONCLUSION: Stimulation of parasympathetic nerves that innervate the sinoatrial node shortened atrial refractoriness in humans. PMID- 10466487 TI - Parasympathetic inhibition of sympathetic effects on pacemaker location and rate in hearts of anesthetized dogs. AB - INTRODUCTION: The site of impulse origin in the right atrium generally is considered to be a single static locus within the sinoatrial (SA) node. Previous investigators showed that the pacemaker site may shift due to changes in sympathetic or parasympathetic neural activity. We investigated the interactions between sympathetic and parasympathetic influences on the site of impulse initiation in the right atrium in anesthetized dogs. METHODS AND RESULTS: We determined the site of impulse initiation and the spread of excitation over the anterior and posterior regions of the right atrium by a matrix of 48 unipolar recording electrodes. We assessed the spread of excitation at 3-msec intervals by constructing isochronal activation sequence maps. Sympathetic stimulation increased the frequency of atrial excitation (i.e., the heart rate), but also shifted the earliest activation region (EAR) from a locus in the SA node to a locus in the superior vena cava (the superior pacemaker site). Vagus stimulation decreased the heart rate and shifted the EAR to a lower site in the SA node or a site in the inferior right atrium along the sulcus terminalis (the inferior pacemaker site). A short period of vagus stimulation during a more prolonged sympathetic stimulation elicited a larger decrease in rate than did vagus stimulation alone and shifted the EAR from the superior site to the SA node or to the inferior site. After atropine, combined stimulation shifted the EAR to the superior site, but propranolol did not change EAR location. CONCLUSION: Our results suggest that parasympathetic activity predominates over sympathetic activity not only on heart rate, but also on the location of the EAR in the anesthetized dog. PMID- 10466489 TI - Electrical restitution, critical mass, and the riddle of fibrillation. PMID- 10466488 TI - Relation between cellular repolarization characteristics and critical mass for human ventricular fibrillation. AB - INTRODUCTION: The critical mass for human ventricular fibrillation (VF) and its electrical determinants are unclear. The goal of this study was to evaluate the relationship between repolarization characteristics and critical mass for VF in diseased human cardiac tissues. METHODS AND RESULTS: Eight native hearts from transplant recipients were studied. The right ventricle was immediately excised, then perfused (n = 6) or superfused (n = 2) with Tyrode's solution at 36 degrees C. The action potential duration (APD) restitution curve was determined by an S1 S2 method. Programmed stimulation and burst pacing were used to induce VF. In 3 of 8 tissues, 10 microM cromakalim, an ATP-sensitive potassium channel opener, was added to the perfusate and the stimulation protocol repeated. Results show that, at baseline, VF did not occur either spontaneously or during rewarming, and it could not be induced by aggressive electrical stimulation in any tissue. The mean APD at 90% depolarization (APD90) at a cycle length of 600 msec was 227+/-49 msec, and the mean slope of the APD restitution curve was 0.22+/-0.08. Among the six tissues perfused, five were not treated with any antiarrhythmic agent. The weight of these five heart samples averaged 111+/-23 g (range 85 to 138). However, after cromakalim infusion, sustained VF (> 30 min in duration) was consistently induced. As compared with baseline in the same tissues, cromakalim shortened the APD90 from 243+/-32 msec to 55+/-18 msec (P < 0.001) and increased the maximum slope of the APD restitution curve from 0.24+/-0.11 to 1.43+/-0.10 (P < 0.01). CONCLUSION: At baseline, the critical mass for VF in diseased human hearts in vitro is > 111 g. However, the critical mass for VF can vary, as it can be reduced by shortening APD and increasing the slope of the APD restitution curve. PMID- 10466490 TI - Characteristics of ischemic and peri-ischemic regions during ventricular fibrillation in the canine heart. AB - INTRODUCTION: Although premature beats originating in areas of ischemia have been shown to be important in initiating ventricular fibrillation (VF), the participation of the ischemic zone in maintenance of VF has not been investigated. METHODS AND RESULTS: Ten normal dogs underwent induction of two separate episodes of VF, before and 10 minutes after left anterior descending coronary artery ligation. Ischemic VF was allowed to occur spontaneously or was induced by burst pacing after 10 minutes of ischemia. Unipolar epicardial electrograms were recorded using an 8 x 14 plaque electrode array (interelectrode distance 2.5 mm) placed over the anterior wall. Activation during VF was characterized by VF cycle length (CL) and wavefront organization based on linking analysis of epicardial activation directions at adjacent sites. Individual plaque sites were separated into regions based on electrogram morphology during ischemia: R1 = no ischemia; R2 = mild-to-moderate ischemia (minor ST elevation and QRS widening); and R3 = severe ischemia (marked ST elevation and QRS widening). Percent conduction block was calculated based on the percent of cycles during which sites were not activated during VF. There were no significant differences noted in mean CL or mean percent conduction block in the peri ischemic region R1 compared to the same region under nonischemic (control) conditions. During ischemia, the mean CL was noted to increase in R2 from 111+/ 14 msec (control) to 128+/-29 msec (ischemia) and in R3 from 113+/-14 msec (control) to 150+/-42 msec (ischemia) (P < 0.05). The percentage conduction block in R2 increased from 6%+/-11% (control) to 14%+/-16% (ischemia) and in R3 from 4%+/-6% (control) to 44%+/-21% (ischemia) (P < 0.05). Linking analysis revealed no significant changes in VF organization at distances of 2.5 mm in regions R1 and R2 under both control and ischemic conditions. Premature beats initiating fibrillation originated at the border between the normal and mildly ischemic zones. CONCLUSIONS: (1) Some VF characteristics are altered in ischemic regions including a longer VFCL and greater percentage of functional block. (2) VF characteristics are unchanged in immediately adjacent nonischemic myocardium. (3) Although the ischemic zone may be involved in the initiation of VF and has unique activation characteristics during VF, it does not affect VF characteristics in the adjacent nonischemic zone, suggesting that it may not play a major role in VF maintenance. PMID- 10466491 TI - Repetitive electrical remodeling by paroxysms of atrial fibrillation in the goat: no cumulative effect on inducibility or stability of atrial fibrillation. AB - INTRODUCTION: Episodes of atrial fibrillation (AF) are known to cause both a rapid reduction in atrial refractoriness (atrial electrical remodeling) and a more delayed increase in AF stability in the chronic goat model. The aims of this study were to examine (1) the hypothesis that an AF-induced increase in AF stability might be due to a mechanism with a longer onset and offset than that of changes in refractoriness and (2) the possibility that repeated paroxysms of maintained AF might cause a cumulative increase in AF stability independent of changes in atrial refractoriness. METHODS AND RESULTS: AF was maintained by rapid atrial pacing in seven goats for three consecutive 5-day periods, each separated from each other by 48 hours of sinus rhythm. Assessments of atrial refractory periods, conduction velocity, AF inducibility, and duration of individual episodes of AF were attempted at intervals throughout the protocol. Forty-eight hours of sinus rhythm was just sufficient for refractoriness changes to fully reverse in all goats, with no evidence of any "residual" increase in AF inducibility. There was no significant difference among any of the three periods of pacing-maintained AF with regard to time to develop episodes of AF of 60 second duration (22.1+/-13, 23.8+/-16, and 30.3+/-29 hours), 1-hour duration (56.6+/-28, 61.3+/-31, and 60.1+/-32 hours), or 24-hour duration (84.0+/-31, 87.0+/-33, and 83.5+/-32 hours). CONCLUSION: There is no evidence for a cumulative effect of AF paroxysms on AF inducibility or stability independent of changes in refractoriness. These findings highlight the importance of atrial refractoriness as a potential target for antiarrhythmic strategies aimed at inhibiting the self-perpetuation of AF. PMID- 10466492 TI - "Atrial fibrillopathy": is atrial fibrillation really self-perpetuating? PMID- 10466493 TI - A case of catheter ablation of accessory atrioventricular connection between the right atrial appendage and right ventricle guided by a three-dimensional electroanatomic mapping system. AB - A 12-year-old girl was referred to our institution because of frequent episodes of AV reciprocating tachycardia. Ventriculoatrial and AV intervals were relatively long along the tricuspid annulus. Earliest retrograde atrial activation was recorded at the mid-portion of the right atrial appendage, 7 mm from the tricuspid annulus. The CARTO electroanatomic mapping system was very useful for providing accurate spatial orientation of the accessory connection. Complete ablation of this connection required multiple radiofrequency energy applications over an extensive area because of the multicomponent structure of the connection. PMID- 10466494 TI - Structure and location of accessory muscular atrioventricular connections. PMID- 10466495 TI - The M cell: its contribution to the ECG and to normal and abnormal electrical function of the heart. AB - The discovery and characterization of the M cell, a unique cell type residing in the deep layers of the ventricular myocardium, has opened a new door in our understanding of the electrophysiology and pharmacology of the heart in both health and disease. The hallmark of the M cell is the ability of its action potential to prolong much more than that of other ventricular myocardial cells in response to a slowing of rate and/or in response to agents that act to prolong action potential duration. Our goal in this review is to provide a comprehensive characterization of the M cell, its contribution to transmural heterogeneity, and its role in the normal electrical function of the heart, in the inscription of the ECG (particularly the T wave), and in the development of QT dispersion, T wave alternans, long QT intervals, and cardiac arrhythmias, such as torsades de pointes. Our secondary goal is to address the controversy that has arisen relative to the functional importance of the M cell in the normal heart. The controversy derives largely from the failure of some investigators to demonstrate transmural heterogeneity of repolarization in the dog in vivo under control conditions and after administration of quinidine. The inability to demonstrate transmural heterogeneity under these conditions may be due to the use of bipolar recording techniques that, in our experience, seriously underestimate transmural dispersion of repolarization (TDR). The use of sodium pentobarbital and alpha chloralose as anesthesia also is problematic, because these agents reduce or eliminate TDR by affecting a variety of ion channel currents. Finally, attempts to amplify transmural dispersion of repolarization with an agent such as quinidine must take into account that relatively high concentrations can result in effects opposite to those desired due to drug inhibition of multiple ion channels. These observations may explain the inability of earlier studies to detect the M cell. PMID- 10466496 TI - Mechanisms and clinical implications of atypical atrial flutter. PMID- 10466497 TI - Seemingly complex QRS alternation: what is the mechanism? PMID- 10466498 TI - Ventricular tachycardia in a patient with cardiac lipoma. PMID- 10466499 TI - ESCWGA/NASPE/P experts consensus statement: living anatomy of the atrioventricular junctions. A guide to electrophysiologic mapping. Working Group of Arrhythmias of the European Society of Cardiology. North American Society of Pacing and Electrophysiology. AB - Current nomenclature for the AV junctions derives from a surgically distorted view, placing the valvar rings and the triangle of Koch in a single plane with anteroposterior and right-left lateral coordinates. Within this convention, the aorta is considered to occupy an anterior position, whereas the mouth of the coronary sinus is shown as being posterior. Although this nomenclature has served its purpose for the description and treatment of arrhythmias dependent on accessory pathways and AV nodal reentry, it is less than satisfactory for the description of atrial and ventricular mapping. To correct these deficiencies, a consensus document has been prepared by experts from the Working Group of Arrhythmias of the European Society of Cardiology and from the North American Society of Pacing and Electrophysiology. It proposes a new, anatomically sound, nomenclature that will be applicable to all chambers of the heart. In this report, we discuss its value for description of the AV junctions and establish the principles of this new nomenclature. PMID- 10466500 TI - Transmission of classical swine fever virus by artificial insemination. AB - Classical swine fever (CSF) virus was introduced into an artificial insemination centre during the CSF epizootic of 1997-1998 in the Netherlands. The risk of further spread of CSF virus via contaminated semen was recognised, but could not be assessed because scientific data on this issue were not available. An animal experiment was performed to determine whether CSF virus could be transmitted via artificial insemination with contaminated semen. Three boars were inoculated with a CSF virus field isolate and from Day 5 till Day 18 thereafter, ejaculates were collected and prepared for insemination. Ruttish sows were inseminated with the extended semen from Day 5 till Day 18 after inoculation of the boars. All the inoculated boars remained healthy throughout the experiment and developed CSF neutralising antibodies between 14 and 21 days after inoculation. Virus was isolated from several semen samples collected from 5 till 11 days after inoculation. Two out of six sows inseminated with CSF contaminated semen seroconverted after insemination. All the other sows remained seronegative. In the foetuses of both the seropositive sows, CSF virus was detected at approximately 35 days post insemination. These results demonstrate that adult boars infected with CSF virus can excrete virus with semen and can, subsequently, transmit the virus to sows and their foetuses via artificial insemination. PMID- 10466501 TI - A comparison of extracted proteins of isolates of Dermatophilus congolensis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting. AB - Antigenic diversity within a collection of 18 isolates of Dermatophilus congolensis from different Continents was examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blotting with sera from cattle with clinical dermatophilosis using whole cell extracts obtained by three methods and one extract of extracellular products of D. congolensis. One of the methods involving the release of a lysostaphin-solubilized protein (LSP) of whole cells of D. congolensis revealed a number of discrete and easily identifiable bands in SDS-PAGE which were found suitable for characterizing protein patterns and was, therefore, subsequently used for a comparative analysis of the proteins of all the D. congolensis isolates. Six electropherotypes (ET) of D. congolensis were identified among the 18 isolates using the protein profiles based on the presence of four protein bands at Molecular weights (MW) 62, 28, 17.4 and 16.4 kDa. The ETs were found among isolates from different animal species and from different sources with ET1 consisting of three bovine and two equine isolates; ET2, two bovine and three ovine isolates; ET3, two bovine isolates; ET4, two bovine isolates; ET5, one bovine and one ovine isolates and ET6, two bovine isolates. Immunoblotting of the extracts of D. congolensis isolates with sera from cattle with clinical dermatophilosis infection demonstrated protein bands of MW ranging from 9 kDa to 188 kDa. Sera from chronic dermatophilosis infection demonstrated a 28 kDa protein which was immunodominant in the LSP extracts of all the 18 isolates of D. congolensis tested while sera from mild infections demonstrated mainly the 62 kDa protein in the same extracts. However, many protein bands were demonstrated in surface membrane (TSMP) and extracellular protein extracts with sera from only mildly infected animals. The protein patterns observed in all isolates of D. congolensis revealed global antigenic similarities and distinct differences among isolates which could not be associated with either geographic, climatic or host factors. Also sera from infected animals from endemic regions of dermatophilosis could not differentiate isolates of D. congolensis. This suggests the possibility that such sera must have come from animals that had been infected by a multitude of D. congolensis strains present in the herd environment and strains an animal could have come across during the 'ritual' annual cross-country migration of the cattle herds. PMID- 10466502 TI - Salmonella infections in finishing pigs in The Netherlands: bacteriological herd prevalence, serogroup and antibiotic resistance of isolates and risk factors for infection. AB - Salmonellae are wide spread in man and animals world wide and are of increasing significance as causative agents of foodborne diseases in man. The European Union, national authorities and the pig industry are therefore more and more interested in the Salmonella status of the pig population. The aim of this study was to estimate the bacteriological prevalence of Salmonella in finishing pig herds, the serogroup and the resistance to antibiotics of the isolated Salmonellae and a preliminary risk analysis of factors associated with infection. For this, 317 finishing pig herds were randomly selected from a database containing 1500 herds in the southern part of the Netherlands. In each herd 24 samples of fresh faeces were collected from two compartments with pigs close to market weight. Per compartment 12 samples of faeces were pooled into one pooled sample. Pooled samples were cultured in duplicate. Salmonella spp. were recovered from 71 out of 306 herds (23%) in which two compartments could be sampled. A total of 108 isolated Salmonella's were serotyped: 71 serogroup B, 3 serogroup C1, 6 serogroup C2, 22 serogroup D1, and 6 isolates neither serogroup B, C or D1. Of a total of 115 Salmonella isolates tested, none were resistant to colistin, enrofloxacin, flumequin or gentamicin. Automated liquid feeding of by-products, and membership of an Integrated Quality Control (IQC) production group were associated with a decreased risk of infection, while use of trough feeding was associated with an increased risk of infection. It is necessary to test these presumed risk factors in intervention studies to evaluate their potency to reduce the Salmonella prevalence in finishing pigs and thereby reduce the risk of Salmonellosis in people consuming pork. PMID- 10466503 TI - In vivo pathogenicity and resistance to phagocytosis of Streptococcus equi strains with different levels of capsule expression. AB - The glossy non-encapsulated strain of Steptococcus equi, NCTC 9682, was compared with the matt strain Hidaka/95/2 which expresses a medium sized capsule and with the mucoid CF32 which expresses a large sized capsule in phagocytosis assays and for virulence in inoculated horses. The three strains, NCTC 9682, Hidaka /95/2 and CF32 produced 2.0, 3.1, and 5.3 mg/g wet cells respectively after 3 h incubation, but similar amounts of M-like proteins, cytotoxin and mitogen. NCTC 9682 showed no resistance to phagocytosis by equine neutrophils regardless of the presence of opsonin while strains Hidaka /95/2 and CF32 showed almost complete resistance to phagocytosis. Furthermore, NCTC 9682 produced no clinical disease although it infected the guttural pouch and caused seroconversion. Typical strangles with guttural pouch invasion was observed in all horses infected with encapsulated strains. PMID- 10466504 TI - The association of Flavobacterium columnare strains of high and low virulence with gill tissue of black mollies (Poecilia sphenops). AB - The ability of a high virulence strain (AJS 1) and a low virulence strain (AJS 4) of Flavobacterium columnare (Flexibacter columnaris) to attach to the gills of black mollies (Poecilia sphenops) was investigated. For that purpose, two groups of 25 black mollies each were immersed in a bath containing 10(6) CFU/ml of F. columnare AJS 1 or AJS 4. At regular intervals from 1 to 12 h after the contact infection, fish were sacrificed and gills, skin, spleen and heart were sampled for bacteriology. Samples of the gills were taken for immunohistochemical and electron microscopic examination. Bacteriological examination proved that the number of gill-associated F. columnare was higher for AJS 1 than for AJS 4. Strain AJS 1 was isolated from the heart and spleen of 6 and 1 of the 16 examined animals, respectively. Strain AJS 4 was not isolated from the internal organs of any fish. When examined immunohistochemically, strain AJS 1 was found closely associated with gill epithelium whereas this was not the case for strain AJS 4. The adherence of bacteria to the gill tissue challenged with the virulent strain AJS 1 was also clearly demonstrated using scanning and transmission electron microscopy. These results indicate that adhesion of F. columnare to the gill tissue constitutes an important step in pathogenesis. PMID- 10466505 TI - Distribution of Salmonella in swine herds in Quebec. AB - Five porcine finishing units, previously identified as contaminated by Salmonella, were sampled to identify possible sources of contamination and to study the distribution of Salmonella within the herds. A total of 208 environmental samples were taken and 87 samples (42%) were found contaminated by Salmonella spp. Salmonella was recovered from several types of samples. Among these, fecal material from pens, building environment such as doors, floors, ventilation units, dust and farm accessories were most often found positive. Some of the flies and rodents were also positive. Two of the finishing units were part of an integrated production system and the prevalence and distribution of Salmonella spp. at different production steps of the integrated facilities were studied. Forty-one farms were sampled and a total of 1923 faecal samples in randomly selected pens were analysed. One hundred and fifty-one samples (7.9%) were positive for Salmonella spp. Among the farms sampled, 70.7% (29/41) were positive for isolation of Salmonella. The different levels in the integrated production were unevenly contaminated. Replacement sow (15.9%) and finishing unit for gilts (21.9%) were the most contaminated levels. Ten serotypes of Salmonella (n = 132) were identified in the production pyramid with a predominance of Salmonella Derby (37.1%) and Salmonella Typhimurium (34.1%). Pulse Field Gel Electrophoresis analysis of the various isolates from serotypes and Salmonella Typhimurium, Salmonella Derby and Salmonella Anatum showed no variation in the genetic profiles, within each serotype, suggesting a vertical contamination throughout the different production steps. PMID- 10466506 TI - Genotypic prevalence of F4 variants (ab, ac, and ad) in Escherichia coli isolated from diarrheic piglets in Korea. AB - A total of 812 Escherichia coli strains isolated from diarrheic piglets were tested for the presence of the F4 (K88) variant (ab, ac and ad) gene by the polymerase chain reaction. Forty four (5.4%) of the 812 E. coli strains carried genes for F4. Among the 44 isolates known to carry genes for F4, 42 (96%) isolates contained genes for F4ac and 2 (4%) isolates contained genes for F4ab. None of the E. coli strains carried genes for F4ad. Our data show that F4ac is the predominant F4 variant associated with diarrhea in piglets in Korea. PMID- 10466507 TI - The influence of symptoms, age, comorbidity and cancer site on physical functioning and mental health of geriatric women patients. AB - This article focuses on how cancer site, age and comorbid conditions of elderly women cancer patients influence their reporting of symptoms; how these variables in turn impact their physical functioning and mental health; and whether there are differences according to the site of the cancer. For the study, a sample of 299 women, age 65 and above, was selected from an on-going longitudinal study of cancer patients recruited from six cancer treatment centers in Michigan and who were recently diagnosed with breast, colon or lung cancer. Age, comorbidity and cancer site were predictors of symptom severity. Age, comorbidity, symptom severity and cancer site were significant predictors of physical functioning, while mental health scores were only predicted by symptom severity. Lung cancer patients reported greater losses in physical functioning than either breast or colon cancer patients, however there were no significant differences in average mental health scores among the three cancer sites. A systematic and comprehensive approach to self-care management of symptom distress provided by oncologists and other health care professionals may enable patients, especially elderly patients with comorbid conditions, to successfully alleviate or reduce the impact of symptoms on their lives. PMID- 10466508 TI - The association between extent of employment and hypertension among women participants of the Second National Health and Nutrition Survey. AB - The association between extent of employment and hypertension was examined among 3,824 women participants of the Second National Health and Nutrition Examination Survey. Women in three employment groups (full-time, part-time, and short-term) were compared to homemakers. In covariate-adjusted, logistic regression models, women employed full-time had a somewhat lower, but not significantly different, prevalence of hypertension (OR = 0.8, 95% CI = 0.7, 1.1), whereas both part-time (OR = 0.6, 95% CI = 0.4, 0.7) and short-term (OR = 0.5, 95% CI = 0.4, 0.7) workers had significantly lower prevalences. Associations did not tend to vary by age, ethnicity, or occupation. Employment does not appear to increase the risk of hypertension in women; in fact, moderate levels were associated with lower risk. PMID- 10466509 TI - The relationship between asthma and smoking during pregnancy. AB - Asthma is the most common and potentially serious medical problem encountered in pregnancy. The prevalence of asthma during pregnancy is generally quoted as being between 1% and 4% which is much lower than general population estimates. The aim of this analysis was to estimate the prevalence of asthma and smoking during pregnancy; and to describe the relationship between the two. A cross-sectional random sample survey of recently delivered Western Australian mothers was carried out in 1995 and 82% of mothers responded. In total 21.3% of women reported ever having asthma at some stage in their life and 12.4% currently had asthma. These results are consistent with general population estimates. Overall 22.7% of women smoked at some stage in pregnancy, which is consistent with the limited comparison data available. Current asthmatics were more likely to smoke during pregnancy than non-asthmatics, although this difference was not statistically significant. Continued vigorous efforts are required to discourage all women and girls from taking up smoking and to encourage those who smoke to quit. This is particularly pertinent for asthmatic women who are pregnant or planning a pregnancy. PMID- 10466510 TI - At-risk behaviors with regard to HIV and addiction among women in prison. AB - The bulk of studies pertaining to addiction among delinquents have been conducted on male subjects. However, the few studies examining female inmates show that a significant proportion of them present an addictive disorder. Furthermore, the HIV infection rate is higher among these women than among incarcerated men. This study attempts to verify if women presenting a combination of criminal and addictive behaviors are at a higher risk of developing an earlier and a more severe delinquency than other delinquent women. Another goal is to determine whether women showing this comorbidity present a higher incidence of HIV-related risk behaviors. The study was conducted on a sample of 210 women from the Montreal detention center. It shows that addicted inmates present earlier onsets of both drug use and criminal behaviors compared to other female inmates. Addicted women also exhibit significantly more HIV-related risky behaviors, both in their drug use and in their sexual practices. PMID- 10466511 TI - Urinary incontinence in working women: an exploratory study. AB - Women who work for a large academic center were surveyed about urinary incontinence. The response rate was 57%. Of the 1113 usable questionnaires, 232 (21%) indicated that urinary incontinence occurred at least monthly. The average age of incontinent women was 45 years (SD = 10 years). Incontinent women were significantly older and had a higher body mass index than continent women. There were also ethnic differences between incontinent and continent groups, although ethnicity was not a predictor of incontinence. Only a third of the women thought incontinence was an important problem to resolve and 46% reported incontinence to their physician or nurse. Forty percent reported that they did not know if the incontinence could be improved, but 81% wanted to learn more about incontinence. PMID- 10466512 TI - Community attitudes and mammography use: does it really matter what other people think? AB - This study examined whether community characteristics, particularly community attitudes regarding mammography use, are associated with women's use of mammography in rural communities. Forty communities in predominantly rural areas of Washington State were selected for inclusion in this study based on their size and distance from an urban center. Characteristics of the communities were assessed as were characteristics of women living in the communities. From each community, random samples of 352 women between 50 and 80 years old participated by completing a telephone survey that included questions on a variety of topics, including their use of mammography. Logistic regression analyses revealed community of residence to be a significant predictor of individual women's mammography use after adjusting for individual level predictors of mammography use including age, education, employment, marital status, financial situation, and ease of access to medical services. An examination of the influence of community characteristics revealed women living in communities with supportive community attitudes towards mammography use report higher levels of mammography use than do women living in communities with less supportive attitudes. The presence or absence of male or female physicians or of mammography facilities in a local community was not associated with statistically significant effects on women's mammography use. Community attitudes are associated with mammography use in rural communities. Public health interventions that change community attitudes may have effects that extend beyond the people directly contacted by these interventions. PMID- 10466513 TI - The acceptability of the female condom among substance-using women in Washington, DC. AB - This research is based on structured interviews, semi-structured interviews, and informal firsthand observation of women residents of Washington, DC who used crack and/or injected drugs during the previous 30 days. The study entailed introducing these women to the female condom, exposing them to an HIV risk reduction intervention teaching them how to use it and how to negotiate its use with their sexual partner(s). Women were tested for HIV and asked to return one week later for their results. They were asked to try the female condom within that first week. Upon returning for their tests results, ethnographers discussed with them their experiences with the female condom. They were reinterviewed for follow-up three months later to assess changes in behavior from baseline as well as their longer term experiences with and opinions of the female condom. The data presented in this paper are based on the interviews conducted one week after baseline. Of particular interest and concern to this research were: women's perceptions of the female condom prior to and subsequent to using it, women's partners' perceptions of the female condom after being introduced to it, and potential barriers to use. In all, 131 women, mostly African-American, took part in this study, which was conducted during the winter of 1997-1998. PMID- 10466514 TI - How health care providers help battered women: the survivor's perspective. AB - This qualitative study aimed to describe, from the perspective of domestic violence survivors, what helped victims in health care encounters improve their situation and thus their health, and how disclosure to and identification by health care providers were related to these helpful experiences. Semi-structured, open-ended interviews were conducted with a purposeful sample of survivors in the San Francisco Bay Area. Data were analyzed using constant comparative techniques and interpretative processes. Twenty-five women were interviewed, the majority being white and middle-class, with some college education. Two overlapping phenomena related to helpful experiences emerged: (1) the complicated dance of disclosure by victims and identification by health care providers, and (2) the power of receiving validation (acknowledgment of abuse and confirmation of patient worth) from a health care provider. The women described a range of disclosure and identification behaviors from direct to indirect or tacit. They also described how-with or without direct identification or disclosure-validation provided "relief," "comfort," "planted a seed," and "started the wheels turning" toward changing the way they perceived their situations, and moving them toward safety. Our data suggest that if health care providers suspect domestic violence, they should not depend on direct disclosure, but rather assume that the patient is being battered, acknowledge that battering is wrong, and confirm the patient's worth. Participants described how successful validation may take on tacit forms that do not jeopardize patient safety. After validating the patient's situation and worth, we suggest health care providers document the abuse and plan with the patient for safety, while offering ongoing validation, support, and referrals. PMID- 10466515 TI - Usefulness of dual SPECT with Tc-99m pyrophosphate and Tl-201 to predict further events after acute myocardial infarction with single-vessel coronary artery disease. AB - PURPOSE: This study was undertaken to determine whether the findings of dual SPECT with Tc-99m pyrophosphate (PYP) and Tl-201 were predictive of further cardiac events after acute myocardial infarction. METHODS: The authors evaluated 88 patients with acute myocardial infarction who underwent dual SPECT for single vessel coronary artery disease. RESULTS: Twenty-nine patients showed overlapping of Tc-99m PYP and Tl-201 in the same location (overlap-positive group), and 59 patients had no overlap (overlap-negative group). In patients in the overlap positive group, the incidence of subsequent events was significantly higher than in patients in the overlap-negative group (P < 0.001). In the overlap-positive group, the number of overlap segments in patients with further events was significantly greater than that in patients without further events (P < 0.005). CONCLUSIONS: Areas with overlapping of Tc-99m PYP and Tl-201 may contain jeopardized myocardium. These results suggest that patients who have a Tc-99m PYP and Tl-201 overlap-negative scan are a low risk group, whereas patients who have more overlapping segments may require catheterization and revascularization. Thus simultaneous SPECT imaging with Tc-99m PYP and Tl-201 might be useful to identify patients with greater ischemic risk after acute myocardial infarction. PMID- 10466516 TI - Perfusion and blood pool scintigraphy for diagnosing soft-tissue arteriovenous malformations. AB - PURPOSE: Six arteriovenous malformations were assessed to determine the effectiveness of perfusion and blood pool scintigraphy to diagnose arteriovenous malformations of soft tissue. METHODS: After injection of Tc-99m RBC or Tc-99m DTPA-HSA, dynamic perfusion and early and delayed blood pool images were analyzed. RESULTS: Four of six arteriovenous malformations showed increased activity on perfusion images and slightly increased activity on blood pool images; the remaining two arteriovenous malformations showed increased activity on perfusion and normal activity on blood pool studies. CONCLUSIONS: This expansion of the authors' previous studies of hemangiomas shows that perfusion and blood pool scintigraphy are useful methods to differentiate arteriovenous malformations from various types of hemangiomas. PMID- 10466517 TI - Preoperative lymphoscintigraphy in the evaluation of squamous cell cancer of the vulva. AB - PURPOSE: A pilot study was undertaken to determine the lymphatic drainage of vulvar cancer using cutaneous lymphoscintigraphy. METHODS: Six patients with biopsy-proved T1 squamous cell cancer of the vulva were studied using 0.4 to 0.6 mCi Tc-99m HSA. Planar imaging was performed after patients received intradermal injections of Tc-99m HSA in a total volume of 0.4 ml at four sites around the vulvar lesion. RESULTS: Tumor locations included two midline lesions and three anterior third lesions. One tumor was located in the midthird of the labia majora. There was no clinically suspicious inguinal adenopathy in any patient. Based on classic anatomic descriptions of cutaneous lymphatic drainage, all but one patient would have been predicted to have drainage to both inguinal nodal basins. Cutaneous lymphoscintigraphy was successful in all six patients. Unilateral drainage was shown in five of six patients. Only one patient had bilateral inguinal drainage, and her tumor was located in the left anterior third of the labia minora. CONCLUSIONS: Cutaneous lymphoscintigraphy with Tc-99m HSA is easily performed and may be potentially useful in defining lymphatic basins at risk in squamous cell cancer of the vulva. PMID- 10466518 TI - Tl-201 and Ga-67 scintigraphy in non-Hodgkin's lymphoma. AB - Ga-67 scintigraphy is routinely used to stage and monitor non-Hodgkin's lymphoma (NHL). It is highly sensitive in high-grade NHL but less so in intermediate- and low-grade NHL. Several studies have reported the use of Tl-201 in the low and intermediate grades of NHL and found it superior in low-grade NHL. In this study, the authors evaluated the utility of combined Ga-67 and Tl-201 scintigraphy in low, intermediate, and unusual types of NHL. Combined Tl-201 and Ga-67 scintigraphy were done in 33 patients (18 women, 15 men; age range, 21-91 years; mean age, 56 years). Tl-201 and Ga-67 had similar overall patient sensitivity in the 33 patients studied. However, the use of both agents increased the overall patient sensitivity from 67% (for Ga-67 only) to 82% and improved the overall site detection from 59 positive sites with Ga-67 to 81 abnormal sites with both Tl-201 and Ga-67. The combined use of Tl-201 and Ga-67 scintigraphy in low- and intermediate-grade NHL resulted in increased disease and site detection and is beneficial for clinical follow-up. PMID- 10466519 TI - Tc-99m dextran and Tc-99m HIG findings in patients with ulcerative colitis. AB - PURPOSE: Various radionuclide methods have been studied for the evaluation of the disease activity and extent of ulcerative colitis and other protein-losing enteropathies. Recently, Tc-99m dextran and Tc-99m human immunoglobulin (HIG) have been used to detect inflammation and protein loss into the intestine, but only a few studies have been reported with these agents. MATERIALS AND METHODS: In this study, Tc-99m dextran and Tc-99m HIG were used to evaluate disease activity and extent in patients with ulcerative colitis. These agents were used in 12 patients with active disease and in five patients in remission, and five healthy control participants also were included. RESULTS: Large bowel activity was detected in 11 of the 12 patients with active ulcerative colitis using Tc-99m dextran and in 10 patients using Tc-99m HIG. Fifty-eight bowel segments were found to be active with endoscopy, 39 with Tc-99m dextran, and 31 with Tc-99m HIG. No intestinal activity was detected in the control participants. Grade 1 activity localization in the large bowel was detected in three patients with ulcerative colitis in remission using Tc-99m dextran and in one patient using Tc 99m HIG. CONCLUSION: Tc-99m dextran is more sensitive for detecting disease activity and extent than is Tc-99m HIG. PMID- 10466520 TI - Normal variants of a photon-deficient area in the lower sternum demonstrated by bone SPECT. AB - PURPOSE: A highly variable normal appearance of the sternal area makes interpretation difficult on planar bone imaging. These normal variants occur especially in the area above the xiphoid process of the sternum in which an oval photopenic area on anterior planar images may be seen. This finding is most likely caused by localized incomplete fusion. METHODS: Of a group of 188 patients, 35 patients (33 men, 2 women; age range, 30 to 85 years; mean age, 57 years) were examined with sternal SPECT and planar bone imaging. These patients, who had no history of sternal trauma, surgery, radiation, or infection, were referred for bone scan for metastatic work-up (n = 20), back pain (n = 6), and other conditions (n = 9). Thus 11 of 35 (31%) patients had a SPECT study that showed a photopenic area in the lower sternum. RESULTS: The incidence of this normal variant was higher than the 2% to 7.7% previously reported, indicating improved detection by SPECT. The normal variant appears to be less apparent on planar bone images and is seen more clearly on SPECT images. CONCLUSION: Differentiation from malignancy appears to be related to lesion symmetry, location, midline, and evenly distributed radioactivity surrounding the edge of the photopenic area. PMID- 10466521 TI - Determination of extent and activity with radionuclide imaging in Erdheim-Chester disease. AB - Erdheim-Chester disease usually involves the diaphyseal and metaphyseal regions of tubular bones and various visceral organs. A 56-year-old woman presented with the histologically confirmed diagnosis of Erdheim-Chester disease. A Tc-99m MDP bone scan revealed the entire extent of the skeletal disease and showed unusual involvement of the epiphyses and axial skeleton. In addition to MRI, a Ga-67 citrate scan including SPECT showed extensive soft-tissue infiltration of different organs. Both Tc-99m MDP and Ga-67 scintigraphy are useful tools in determining the distribution of this rare disease. PMID- 10466522 TI - Somatostatin receptor scintigraphy for early detection of regional and distant metastases of medullary carcinoma of the thyroid. AB - Three patients are described who had regional and distant metastases of medullary thyroid cancer detected by somatostatin receptor scintigraphy but not by CT; two had minimal disease that was amenable to surgery. The first patient had been followed for 2 years before having a repeated scan and positive CT, with subsequent surgical removal of metastatic paratracheal nodes. The plasma calcitonin level, however, did not approach normal values after surgery, and a third scan showed persistence of focal uptake in the left paratracheal area of the lower neck, whereas CT was negative. At repeated exploration, a tumor mass of medullary carcinoma, embedded in lymphatic tissue, was removed. Nine months after the last surgical procedure, calcitonin and carcinoembryonic antigen levels were normal. The second patient underwent microdissection of the mediastinum and removal of two metastatic nodes that were demonstrable only by the scintigraphic technique. The plasma calcitonin level subsequently became normal. The third patient, with multiple endocrine neoplasia IIB and associated pheochromocytoma, had bony metastatic involvement of the left shoulder, demonstrable initially on somatostatin receptor scintigraphy and subsequently with radioiodinated metaiodobenzylguanidine but not on CT. PMID- 10466523 TI - FDG PET evaluation of residual masses and regrowth of abdominal lymph node metastases from colon cancer compared with CT during chemotherapy. AB - Fluorine-18-2-deoxy-2-fluoro-D-glucose (F-18 FDG) PET may be more suitable for follow-up after cancer treatment than other morphologic approaches, because it reflects tumor viability. A patient with abdominal lymph node metastases from colon cancer was followed by CT and F-18 FDG PET during chemotherapy. F-18 FDG PET tumor images changed in accordance with the clinical progress, whereas CT findings were relatively unchanged. This case clearly shows the utility of F-18 FDG PET for follow-up during cancer chemotherapy. PMID- 10466524 TI - A patient with three kidneys: a correlative imaging case report. AB - A man with three fully developed and well functioning kidneys was studied using correlative imaging. Renal scintigraphy and the renogram not only played a role in identifying the existence of three kidneys but also determined the level of function of each kidney. The use of renal scintigraphy and renography is pivotal in the diagnosis of supernumerary kidneys. An abbreviated review of embryogenesis is also given. PMID- 10466525 TI - False-negative scintigraphy with Tc-99m sestamibi in stage IV neuroblastoma. AB - Tc-99m sestamibi, originally developed for myocardial studies, has been used as a tumor-seeking agent. Recently, the agent also was reported to be a functional tracer to predict multidrug resistance-related p-glycoprotein expression in tumor tissue. The current report presents the authors' experience with sestamibi tumor scintigraphy in a neuroblastoma. Although I-131 MIBG tumor imaging and Tc-99m MDP bone scanning accurately demonstrated the extent of the disease, Tc-99m sestamibi showed no accumulation in primary and metastatic foci. Lack of sestamibi uptake was initially thought to be suggestive of failure to respond to chemotherapy because of p-glycoprotein expression. However, the patient responded well to chemotherapy and complete remission was achieved. The failure of Tc-99m sestamibi to detect a neuroblastoma and the lack of sestamibi accumulation in the tumor may not always be related to chemotherapy resistance. PMID- 10466526 TI - Primary hepatic amyloidosis well delineated by Tc-99m DTPA galactosyl HSA liver SPECT. PMID- 10466527 TI - Gastrointestinal bleeding detected on a routine Tc-99m MDP bone scan. PMID- 10466528 TI - Off-peak detector mimicking ischemia on myocardial scan. PMID- 10466529 TI - Incidental detection of a popliteal pseudoaneurysm on bone scintigraphy. PMID- 10466530 TI - "Elephantine" calf swelling after groin resection and radiation therapy for vulvar carcinoma. PMID- 10466531 TI - Unusual stress fracture in an athlete. PMID- 10466532 TI - Increased F-18 FDG uptake in the stomach. PMID- 10466533 TI - Crossed cerebellar diaschisis demonstrated by brain SPECT with Tc-99m MAA in a child with an intracardiac right-to-left shunt. PMID- 10466534 TI - Bone scintigraphy of Madura foot. PMID- 10466535 TI - Stress fractures of the thoracic spine transverse processes in a water skier. PMID- 10466536 TI - Tc-99m MAG3 scintigraphy in a newborn with acute renal vein thrombosis. PMID- 10466539 TI - Current readings in nuclear medicine. PMID- 10466537 TI - Tc-99m pertechnetate scintigraphy to diagnose rectal duplication. PMID- 10466538 TI - Bone metastasis demonstrated as a photopenic area in L3 on bone SPECT. PMID- 10466540 TI - Fluticasone propionate and bronchial hyperresponsiveness in childhood asthma. AB - Bronchial asthma is now agreed as being a chronic inflammatory disease of the airways. Inhaled steroids are widely accepted as a preventive medication in asthmatic patients of all ages and severity. However, the optimal use of inhaled steroids and the important issue of safety and efficacy still remain of concern, particularly in children. Recently, fluticasone propionate (FP) has been developed for use as an inhaled preparation for the treatment of asthma. Because of its high topical potency and increased lipophilicity, it is claimed that FP has an improved risk/benefit compared with other inhaled steroids. In order to evaluate the use of FP in children, we have studied the efficacy of high dose FP (500 microg/day) in asthmatic children. Thirteen children (9 boys and 4 girls), aged 7-17 years (10.8 +/- 2.6), were instructed to use a pressurized metered-dose inhaler connected to a Volumetric spacer. The standard methacholine bronchial challenge test was used as a principal outcome parameter. The PD20, a cumulative dose of methacholine inducing a 20% decrease in FEV1, was measured pre- and post treatment with inhaled FP. After 4 weeks of FP, PD20 significantly increased from 21.6 +/- 14.3 inhalation unit to 106.6 +/- 78.5 inhalation unit (4.9 fold, p = 0.004) reflecting the improvement of airway reactivity. All subjects improved clinically. These results demonstrate that the anti-inflammatory action of FP 500 microg a day for four weeks can markedly reduce bronchial hyperresponsiveness, the basic physiologic abnormality in bronchial asthma. PMID- 10466541 TI - Use of standard radiography to diagnose paranasal sinus disease of asthmatic children in Taiwan: comparison with computed tomography. AB - Paranasal sinus disease and bronchial asthma are frequently associated. Computed tomography imaging is currently the most reliable method for confirming the diagnosis of sinusitis. Due to the cost and amount of radiation during computed tomography, our aim was to analyze whether standard radiography, under computed tomography-control, had a reasonable degree of confidence in the diagnosis of sinusitis. Fifty-three asthmatic patients (42 males and 11 females) with a mean age of 9 years (range 4-14) were enrolled. We evaluated the maxillary sinuses, ethmoidal sinuses, frontal sinuses, and sphenoidal sinuses using standard radiography (Waters' view, Caldwell view, and lateral view) and compared with computed tomography (coronal views), the latter served as a standard. Computed tomography (CT) showed paranasal sinusitis in 58% (31/53) of the asthmatic children. Compared with the results of computed tomography, standard radiography revealed a sensitivity of 81.1% and a specificity of 72.7% for maxillary sinusitis. The sensitivity and specificity for ethmoidal, frontal, and sphenoidal sinusitis were 51.8%, 84.8%; 47.3%, 87.2%; and 40.8%, 93.3%, respectively. In 21 (40%) of the 53 patients, discrepancies were seen between the interpretations of standard radiography c and those of CT scans. In patients with maxillary sinusitis, the correlation between standard radiography and CT was good. However, ethmoidal, frontal, and sphenoidal sinusitis were poorly demonstrated using radiography. Standard radiography can be recommended as a screening method for maxillary sinusitis, but it is not recommended for the diagnosis of other paranasal sinusitis. PMID- 10466542 TI - Pregnancy outcome in Thai patients with systemic lupus erythematosus. AB - The outcome of 48 pregnancies from 42 patients with systemic lupus erythematosus was studied. Their mean age and the duration of the disease were 28.47 and 4.42 years, respectively. The conception occurred when the disease was inactive or quiescent in 45 and active in 3. Four pregnancies were terminated by criminal abortion. Flares occurred in 16 pregnancies. The kidney and mucocutaneous system were the 2 organs that flared most commonly. The fetal outcomes were term delivery in 18 (40.90%), prematurity in 17 (38.64%), spontaneous abortion in 6 (13.64%) and still birth in 3 (6.82%). There was no statistical difference in pregnancy loss and successful delivery between pregnant patients with and without flares. Concerning 35 successful live births, those pregnancies without flares had significantly more full term deliveries (p < 0.02), higher gestational age (p < 0.002) and more birth weight (p < 0.001) than those with flares. Small for gestational age was seen in 20%. Pregnancy with active renal disease had a poor fetal outcome. There were no cases of congenital anomalies or neonatal lupus. Maternal complications were more common in patients with flares. PMID- 10466544 TI - Frequencies of cytotoxic T lymphocyte precursor estimate in three different populations. AB - There is speculation that high cytotoxic T lymphocyte precursor frequencies (CTLpf) correlate with poor clinical outcome of bone marrow/organ transplantation. It is also believed that human umbilical cord blood is immunologically naive, and, therefore cord blood T cells may be less able to mediate graft versus host disease than marrow-derived T cells. CTLpf were determined in peripheral blood mononuclear cells collected from healthy adults, human umbilical cord blood and renal dialysis patients who were randomly selected and entered into this study. A highly sensitive non-radioactive Europium release cytotoxicity assay was optimized and modified to carry out the CTLpf estimation by using the principle of limiting dilution analysis. The results of CTLpf in healthy adults ranged from 1/694 to 1/66,666, median 1/7,339 (n=10); cord blood ranged from 1/1,562 to 1/35,714, median 1/10,162 (n=6) and dialysis patients ranged from 1/1,054 to 1/17,857 median 1/5,208 (n=9). The results demonstrated that there is little difference of CTLpf median values between the groups, but there is a wide variation of CTLpf between individuals within a population. It suggests that this variation should be taken into account when considering CTLpf assay as pre-transplantation cross-match procedure. PMID- 10466543 TI - Clinico-immunopathological alterations of lymph nodes from human immunodeficiency virus-infected patients in northern Thailand. AB - To determine if the immunopathologic alterations of HIV-infected lymph nodes have any correlation with clinical stages in the northern Thai patients, we conducted a comparative analysis of immunopathologic features of lymph nodes between 25 HIV infected patients from various clinical categories and 25 non-HIV individuals of reactive hyperplasia morphology of lymph node biopsies. The risk factors for HIV infection were all heterosexual. The majority of patients in clinical category A (PGL) showed a histopathologic pattern of explosive follicular hyperplasia, while category C (AIDS) patients demonstrated follicular involution and lymphocyte depletion on lymph node sections. Interestingly, weak reactivity for HIV p24 gag protein was detected within the germinal centers and scattering interfollicular lymphocytes in only 20% of the HIV-infected cases. Morphologically, the presence of MGCs was specific for HIV-infected lymph nodes. MGCs (hematoxylin & eosin stain) were found in 64% of the HIV-infected cases, which was significantly different from 4% found in control cases (p = 0.00002). By S-100 immunostaining, MGCs were demonstrated in all HIV-infected lymph node sections, while they were found in 32% of the control lymph nodes. Immunostaining with S-100 protein also revealed the appearance of syncytial ballooning and countable numbers of MGCs. High numbers of MGCs seemed to correlate with histologic and clinical changes. In conclusion, the HIV-infected patients had high numbers of MGCs or syncytia on lymph node sections in early stage and pre-AIDS conditions, which has never been reported before. PMID- 10466545 TI - TT virus infection in intravenous drug users. AB - Our group has investigated 201 intravenous drug users for the presence of TTV DNA by means of polymerase chain reaction (PCR). The majority of the individuals tested were male, their age ranging from 16 to 63 years, and the duration of intravenous drug use from one to 40 years. TTV DNA was present in 62 of the 201 IVDUs (30.8%) with its prevalence on the ascent between the age groups below 20 and those between 21 and 30 years, as well as between the groups below 60 and between 60 to 120 months' duration of drug intake, respectively. When tested again after 9 months, nine IVDU (23.7%) were found TTV negative by PCR hinting at potential immunological clearance. Our control group comprised 200 healthy blood donors, 7% of whom were found to harbor TTV DNA in an age-dependent fashion, as observed with the IVDU. From the liver function tests performed we could not detect any statistically significant difference regarding ALT elevation observed in TTV-positive compared with TTV-negative individuals. To date, TTV does not appear to cause any serious liver disease in the majority of cases examined. PMID- 10466546 TI - Correlation between immunohistochemical and biochemical estrogen receptors in the prognosis of patients with breast cancer. AB - To evaluate the reliability of immunohistochemical estrogen receptor (ER) in the prognosis of patients with breast cancer, 83 primary tumors from patients were studied. Immunohistochemical analysis (IHA) was performed using antibody ER 1D5 (Dako) together with microwave treatment for antigen retrieval. ER values obtained using the biochemical steroid binding assay (polyethyleneglycol method, PEG) were available for comparison. Of all tumors, ER positivity was detected in 44.6% by IHA and 36.1% by PEG method. The concordance between the two methods was 69%. No significant correlation was found between the ER status determined by both methods and clinical stage, tumor size, lymph node status or age of patient at diagnosis. However, we found that the immunohistochemical ER is a superior predictor of early recurrence in patients with primary breast cancer to biochemical ER. The findings in the present study emphasize the clinical benefit of the immunohistochemical ER assay as a measure for prognosis. PMID- 10466547 TI - Comparison study of combined DTPw-HB vaccines and separate administration of DTPw and HB vaccines in Thai children. AB - The safety, immunogenicity and tolerability of two different DTPw-HBV combination vaccines, containing 5 and 10 microg of HBsAg; were investigated in comparison with separate administration of DTPw and HBV (10 microg of HBsAg). A three dose primary vaccination course at 2, 4 and 6 months of age was followed by a booster dose at 18 months. All vaccines were safe and well tolerated. The DTPw-HBV combination vaccine containing 10 microg of HBsAg elicited significantly higher anti-HBs titres than the other two vaccines after the primary and booster vaccination course. All vaccines elicited a high response against the other components. Based on these results, DTPw-HBV (10 microg HBsAg) was the most effective vaccine at this schedule. PMID- 10466548 TI - Acute aortic valvulitis as an initial presentation of systemic lupus erythematosus. AB - Valvular involvement in patients with systemic lupus erythematosus (SLE) is not uncommon but patients rarely present with it. The mitral valve is most commonly involved. We report a 36-year-old man who had an episode of acute fever, arthritis, and acute aortic insufficiency with a small vegetation at the tip of the aortic valve mimicking infective endocarditis, proven later to be due to SLE. SLE should be considered as one of the uncommon causes of acute aortic insufficiency. PMID- 10466549 TI - Caveats regarding the use of control charts. PMID- 10466550 TI - Bugs are not funny syndrome. PMID- 10466551 TI - Gene therapy: a new challenge for infection control. PMID- 10466552 TI - Results of a comprehensive infection control program for reducing surgical-site infections in coronary artery bypass surgery. AB - OBJECTIVE: To evaluate the efficacy of a comprehensive infection control program on the reduction of surgical-site infections (SSIs) following coronary artery bypass graft (CABG) surgery. DESIGN: Prospective cohort study. SETTING: 1,000-bed tertiary-care hospital. PATIENTS: Persons undergoing CABG with or without concomitant valve surgery from April 1991 through December 1994. INTERVENTIONS: Prospective surveillance, quarterly reporting of SSI rates, chlorhexidene showers, discontinuation of shaving, administration of antibiotic prophylaxis in the holding area, elimination of ice baths for cooling of cardioplegia solution, limitation of operating room traffic, minimization of flash sterilization, and elimination of postoperative tap-water wound bathing for 96 hours. Logistic regression models were fitted to assess infection rates over time, adjusting for severity of illness, surgeon, patient characteristics, and type of surgery. RESULTS: 2,231 procedures were performed. A reduction in infection rates was noted at all sites. The rate of deep chest infections decreased from 2.6% in 1991 to 1.6% in 1994. Over the same period, the rate of leg infections decreased from 6.8% to 2.7%, and of all SSI from 12.4% to 8.9%. The adjusted odds ratio (OR) for all SSIs for the end of 1994 compared to December 31, 1991, was 0.37 (95% confidence interval [CI95], 0.22-0.63). For deep chest and mediastinal infections, the adjusted OR comparing the same period was 0.69 (CI95, 0.28-1.71). CONCLUSIONS: We observed significant reductions in SSI rates of deep and superficial sites in CABG surgery following implementation of a comprehensive infection control program. These differences remained significant when adjusted for potential confounding covariables. PMID- 10466553 TI - A pseudoepidemic of postoperative scleritis due to misdiagnosis. AB - OBJECTIVE: To describe a pseudoepidemic of infectious scleritis following eye surgery. METHODS: Retrospective cohort study with selected procedural and laboratory investigations. RESULTS: Twenty-one patients with postoperative scleritis were identified during a 2-month outbreak. Neither an infectious etiology nor a causative pre-, intra-, or postoperative exposure was found. The clinical findings, when carefully reviewed, were consistent with poor surgical wound closure. CONCLUSIONS: The art of clinical diagnosis involves the subjective interpretation of clinical history, physical findings, and laboratory results. A repeated error in the interpretation of clinical findings can simulate an outbreak of disease. Clinicians may be reluctant to concede misdiagnosis. PMID- 10466554 TI - A decade of prevalence surveys in a tertiary-care center: trends in nosocomial infection rates, device utilization, and patient acuity. AB - OBJECTIVE: To evaluate the usefulness of repeated prevalence surveys to determine trends in the rates of nosocomial infections and to detect changes in risk factors (e.g., use of invasive devices) associated with nosocomial infections. PATIENTS AND METHODS: Ten annual prevalence surveys were conducted by trained infection control practitioners between 1985 and 1995 for acute-care patients on the medical, surgical, pediatric, and obstetric-gynecologic services at a 900 bed, tertiary-care, teaching hospital with 750 acute-care beds. The same methods of chart review and concurrent reporting from nursing, the microbiology and clinical laboratory, and the pharmacy were used each year to collect data on the prevalence of nosocomial infections, invasive-device utilization, and abnormal laboratory indicators. Although data were collected on a single day, a period prevalence study approach was used, because charts were reviewed for any infection data occurring within the 7 days prior to the survey. RESULTS: The hospital census for acute care patients, as measured by the prevalence surveys, declined sharply over the 10 years, from 673 to 575 patients (P = .02). However, the medical service census increased from 150 to 188 patients (P = .01). During the same period, there was a significant decrease in the mean length of stay, from 7.3 to 6.0 days (P = .01), and a concomitant increase in the mean diagnosis related-group case-mix index, from 1.03 to 1.24 (P = .001). Overall, nosocomial infection rates remained unchanged over the study period (mean of 9.85 infections per 100 patients), but rates of nosocomial bloodstream infection increased from 0.0% in 1985 to 2.3% in 1995 (P = .05). Nosocomial infection rates were significantly higher on the medical and surgical services than on other services (P<.001). Utilization rates increased significantly for Foley catheters (9.0% to 16.0%, P = .002) and ventilators (5.0% to 8.0%, P = .05). CONCLUSIONS: Despite apparent increases in the severity of illness of our patients, overall rates of nosocomial infection remained stable during a decade of study. Rates of nosocomial bloodstream infection increased, in parallel with National Nosocomial Infection Surveillance System data. We found repeated prevalence surveys to be useful in following trends and rates of infection, device utilization, and abnormal laboratory values among patients at our institution. Such methodologies can be valuable and low-cost components of a comprehensive infection surveillance, prevention, and control program and other potential quality improvement initiatives, because they enable better annual planning of departmental strategies to meet hospital needs. PMID- 10466555 TI - Nosocomial pertussis in healthcare workers from a pediatric emergency unit in France. AB - OBJECTIVE: After pertussis was diagnosed in July 1997 in a 55-year-old nurse (case) from a pediatric emergency unit who had a respiratory illness and paroxysmal cough for 5 weeks, an epidemiological investigation was initiated to determine if other healthcare workers (HCWs) from the same unit also had pertussis. DESIGN: Interviews were conducted to assess symptoms occurring in the previous months. Two sera were collected 2 to 3 months apart for 59 of 61 HCWs of the unit. The IgG response to pertussis toxin was determined using Western blot assay. SETTING: Pediatric emergency unit (61 HCWs) of a 2,500-bed university hospital. RESULTS: There was a total of 10 (5 confirmed and 5 probable) cases of pertussis identified in this outbreak. Nine HCWs (15%) had results suggesting recent or acute pertussis. To avoid transmission to patients and other HCWs, all HCWs with cough were treated for 14 days with erythromycin, and those having acute cough were given a 5-day sick leave. Despite these measures, a new acute pertussis case was identified in a 41-year-old nurse, with a positive culture from nasopharyngeal aspirates. Thus, all HCWs in the unit were prescribed spiramycin for 10 days to prevent any further spread of pertussis. CONCLUSION: Pertussis should be considered a threat to HCWs who are in contact with children. For HCWs, diagnosis of pertussis should be made on a clinical basis, giving greater importance to sensitivity of diagnosis criteria, and on early bacterial identification by culture of the organism or by polymerase chain reaction. RECOMMENDATIONS: In case of pertussis in an HCW, all staff in the unit who have had unprotected and intensive contact with that person should be provided with macrolide treatment to stop any transmission to colleagues and to young patients. Furthermore, the possibility of providing these HCWs with acellular pertussis vaccines warrants further investigation. PMID- 10466556 TI - Two consecutive outbreaks of Stenotrophomonas maltophilia (Xanthomonas maltophilia) in an intensive-care unit defined by restriction fragment-length polymorphism typing. AB - OBJECTIVE: To investigate and control consecutive outbreaks of Stenotrophomonas maltophilia infections in intensive-care-unit (ICU) patients. DESIGN: Epidemiological investigation; restriction fragment-length polymorphism typing by pulsed-field gel electrophoresis (PFGE) of genomic DNA of outbreak strains; institution of infection control measures to limit spread. SETTING: The medical surgical ICU in an 800-bed tertiary-care center in Calgary, Alberta, Canada. RESULTS: S. maltophilia was recovered from 14 ICU patients (12 infected, 2 colonized) between February 1993 and February 1994. Ten of the 14 patient isolates and 1 environmental isolate were available for PFGE typing. Patient isolates from 6 of the first 10 patients were identical. Isolates from the next 3 of 4 patients and an isolate recovered from a ventilator being used by a patient not infected with S. maltophilia also were identical, but different from the first 6. The ventilator isolate was temporally associated with the latter 4 patients. CONCLUSION: Molecular typing allowed us to determine that there were two separate consecutive S maltophilia outbreaks rather than a single protracted outbreak. Recovery of S. maltophilia from patient ventilators and an in-line suction catheter suggests that the organism may have been spread by cross contamination from contaminated equipment or from an environmental source. PMID- 10466557 TI - Vancomycin in Oregon: who's using it and why. AB - OBJECTIVE: To determine the proportion of vancomycin orders that are appropriate according to national guidelines and to identify targets for educational messages. DESIGN: Population-based study of vancomycin use in Oregon during a 3 week period. Survey of pharmacists, prospective flagging of vancomycin orders, and data abstraction from patient charts using standardized forms. SETTING: Nonpsychiatric hospitals in Oregon. RESULTS: Four (6%) of the 66 Oregon hospitals had pharmacy restrictions on initial vancomycin orders. Sixty-four (97%) of the hospitals participated in the study of indications for use; 293 vancomycin orders were reported; 3.8 courses were initiated per 1,000 patient-days. Indications for use were determined for 266 (91%); of these, 159 (60%) were deemed appropriate. Of uses for prophylaxis, empirical treatment of suspected gram-positive infection, and treatment of documented gram-positive infection, 57%, 56%, and 65%, respectively, were appropriate. Of hospitals with <250, 251-475, and >475 licensed beds, 65%, 58%, and 57% of vancomycin orders were appropriate. No single medical specialty accounted for >16% of inappropriate vancomycin use. CONCLUSIONS: Vancomycin was used inappropriately by physicians of many different specialties, in hospitals of all sizes, and in sundry clinical situations. The problem of inappropriate vancomycin use does not lend itself to solution by educational strategies targeted at specific subgroups; restrictions by hospital pharmacies may be required. PMID- 10466558 TI - Breakthrough varicella infection in a healthcare worker despite immunity after varicella vaccination. AB - Although varicella vaccination is recommended for varicella-susceptible healthcare workers (HCWs), breakthrough infection after vaccination is not unusual, especially following household exposures. We report breakthrough varicella in a vaccinated HCW and review the data on breakthrough infection and concerns for the healthcare setting. PMID- 10466559 TI - Varicella-zoster screening and management programs in healthcare facilities in Canada. AB - A 24-question survey on varicella-zoster screening programs and control measures for varicella-zoster infections was sent to 170 Canadian healthcare facilities. The results indicated that 60% of Canadian healthcare facilities have well established preemployment varicella-zoster screening programs for healthcare workers. Overall, 30% of healthcare facilities impose work restrictions for susceptible healthcare workers, 49% use negative-pressure rooms for varicella zoster management, and 28% have a policy to discharge susceptible exposed patients. Large variations exist among Canadian healthcare facilities in their varicella-zoster infection control policies. PMID- 10466560 TI - Nosocomial meningococcemia in a physician. AB - We report the case of a pediatrician who developed meningococcal meningitis after performing endotracheal intubation without protection on a child who was suspected of having meningoencephalitis. This case emphasizes the necessity for healthcare workers who perform high-risk procedures to use personal protection devices (i.e., respirators and protective goggles). Unprotected healthcare workers with high exposure to Neisseria meningitidis should receive chemoprophylaxis. PMID- 10466561 TI - The emergence of resistant strains of Acinetobacter baumannii: clinical and infection control implications. AB - A prospective study was undertaken to determine colonization rates, susceptibility profiles, and outcomes in patients with clinical isolates of Acinetobacter baumannii. Fifty percent of patients became colonized with A. baumannii, and 29% of these patients had clinical and colonizing isolates with discordant susceptibility profiles, without apparent relation to antibiotic use. Barrier infection control measures are necessary to prevent nosocomial transmission. PMID- 10466562 TI - Infection control in gene therapy. AB - Gene therapy is now being studied for the treatment of a wide variety of acquired and inherited diseases. Viruses used as vectors for gene transfer include retroviruses, adenoviruses, vaccinia viruses, adeno-associated viruses, and herpesviruses. These vectors, developed in the laboratory and in animal studies, are now being introduced into the clinical arena Infection control practitioners will be involved invariably in reviewing the use of these agents in their clinics and hospitals. This review summarizes key aspects of the more common vectors and makes recommendations for infection control. PMID- 10466563 TI - OSHA reopens record for TB rule. PMID- 10466564 TI - Recognition performance for four combinations of FM system and hearing aid microphone signals in adverse listening conditions. AB - OBJECTIVE: Children with moderate to severe hearing loss routinely use personal frequency modulated (FM) systems in the classroom to improve the signal to noise ratio of teacher-directed speech with notable success. Attention is now being given to the ability of these children to hear other students via the hearing aid (HA) microphone while using an FM system. As a result, a variety of FM system and HA microphone combinations have been recommended for classroom use. To date, there are no studies regarding the efficacy of these FM/HA combinations. The purpose of this study was to evaluate recognition performance using four FM/HA combinations and to characterize that performance for stimuli received primarily through FM system and HA microphone transmission. DESIGN: Recognition performance for FM system and HA microphone signals was evaluated for two symmetrical and two asymmetrical FM/HA combinations using two commercially available FM systems (one conventional and one FM-precedence circuit). Eleven children (ages 9 to 12) with moderate to severe sensorineural hearing loss and eight children (ages 10 to 11) with normal hearing served as subjects. The two symmetrical FM/HA combinations included: 1) binaural FM system and HA microphone input using the conventional FM system, and 2) binaural FM and HA input using the FM-precedence circuit. The conventional FM system was used for the two asymmetrical combinations and included: 1) binaural FM input and monaural HA input, and 2) FM input to one ear and HA input to the other. Stimuli were 33 consonants presented in the form of nonsense syllables. The stimuli were presented through three loudspeakers representing a teacher and two fellow students in a classroom environment. Speech shaped noise was presented through two additional loudspeakers. RESULTS: In general, no statistically significant differences in recognition performance were found between any of the FM/HA combinations. Mean recognition scores for HA microphone transmission (55%) were significantly poorer than those for FM system transmission (75%). As expected, initial consonants were more easily recognized than final consonants via FM system and HA microphone transmission. However, voiceless consonants were more easily recognized than voiced consonants via HA microphone transmission, which was not predicted on the basis of previous research. CONCLUSIONS: These results suggest that a certain amount of flexibility is present when choosing an FM/HA combination. However, recognition performance via the HA microphones was consistently poorer than performance via FM transmission. Because relevant material also originates from fellow students (e.g., answering teacher-directed questions), input via the HAs is often as important as information originating from the teacher. The results suggest that attempts to improve performance for signals transmitted through the HA microphones in a classroom setting would benefit children with hearing loss. PMID- 10466565 TI - Trial of a two-channel hearing aid (low-frequency compression-high-frequency linear amplification) with school age children. AB - OBJECTIVE: The study was designed to investigate the efficacy of a 2-channel hearing aid with low-frequency compression and high-frequency linear amplification on a group of school-age hearing aid wearers. DESIGN: The study was a single-center, 2-way crossover design in which 25 children (age 6 to 15 yr) were fitted with 2-channel hearing aids for 12 wk and with their own (single channel) hearing aids for 12 wk, refitted according to published protocols. Speech perception in quiet and in noise was measured at the end of each 12 wk period; in addition, questionnaires were given to teachers, parents, and children. RESULTS: Two-channel hearing aids showed significantly higher mean scores for speech perception in noise and significantly higher composite questionnaire scores (reflecting aspects of satisfaction and benefit). Final choice of hearing aids at the end of the study by parents and children also favored the 2-channel device. CONCLUSIONS: The 2-channel hearing aids appear to be an acceptable management option for audiometrically suitable children. The results provide support for the 2-channel design rationale and suggest the need for further trials. PMID- 10466566 TI - Accuracy of hearing aid use time as reported by experienced hearing aid wearers. AB - OBJECTIVE: The purpose of this study was to investigate the relationship between the amount of time that a person reports wearing his or her hearing aid and the actual time that the hearing aid is worn. Although use time data are used in the analysis and interpretation of investigations concerned with auditory perceptual learning and with user satisfaction, the accuracy of self-reported use time has not been fully investigated. DESIGN: The experimental and control group were fit with a hearing aid that has the capability of storing use time data for later analysis. The experimental group was told that the self-reported use time would be verified with a computer analysis of the hearing aid that provides the actual use time. The control group was not informed of the use time validation procedure. The agreement between self-reported and actual use time was compared statistically between groups (knowledge of validation versus no knowledge of validation). RESULTS: The experimental group provided accurate self-reported use time whereas the control group showed a significant difference between actual use time and self-reported use time. CONCLUSIONS: The results may assist in the interpretation of results of previous investigations that have depended on self reported use time and in the design of future investigations. For the clinician, the results indicate that relying on a patient's self-reported hearing aid use time for documentation of satisfaction or signal processing preference may be misleading. PMID- 10466567 TI - Measuring Satisfaction with Amplification in Daily Life: the SADL scale. AB - OBJECTIVE: To develop a self-report inventory to quantify satisfaction with hearing aids. DESIGN: The inventory was developed in several stages. To determine the elements that are most important to satisfaction for most people, we conducted structured interviews and then designed a questionnaire. Hearing aid owners responded to the questionnaire, indicating the relative importance of 14 different elements in their hearing aid satisfaction. Analyses indicated that the elements could be placed into four importance content areas. Trial satisfaction items were designed for each content area and submitted to focus groups to identify highly salient items as well as ambiguous items. A 25-item satisfaction questionnaire then was developed and disseminated to hearing aid owners. Results were obtained from 257 individuals. These data were analyzed to generate the final questionnaire. RESULTS: Fifteen items, divided into four subscales, were selected for the final Satisfaction with Amplification in Daily Life (SADL) questionnaire. The questionnaire yields a Global satisfaction score and a profile of subscale scores, which address Positive Effects, Service and Cost, Negative Features, and Personal Image. A preliminary evaluation of retest stability was conducted with 104 subjects. Ninety percent critical differences for the various scores ranged from 0.9 to 2.0 score intervals on a 7 point scale. CONCLUSIONS: The SADL scale is both brief enough to be clinically acceptable and comprehensive enough to provide a valid assessment of an inherently multidimensional variable. Additional assessment is necessary to refine understanding of its test-retest properties, explore validity issues, and determine clinical, research, and administrative applications of the data. PMID- 10466568 TI - Effects of electrode location and spacing on phoneme recognition with the Nucleus 22 cochlear implant. AB - OBJECTIVE: The objective of this paper was to determine how phoneme identification was affected by the cochlear location and spacing of the electrodes in cochlear implant listeners. DESIGN: Subjects were initially programmed with the full complement of 20 active electrodes, in which each electrode was assigned to represent the output of one filter in the normal SPEAK processor. In the present study several four-electrode processors were constructed by assigning the output of more than one filter to a single electrode. In all conditions speech sounds were still analyzed into 20 frequency bands and processed according to the usual SPEAK processing strategy, but the location and spacing of the four stimulated electrode pairs were varied systematically. In Experiment I, the spacing between stimulated electrodes was fixed at 3.75 mm and the cochlear location of the four electrode pairs was shifted from the most-apical position up to 3.0 mm toward the base in 0.75 mm steps. In Experiment II, the spatial separation between the four electrode pairs (each bipolar-plus-one) was systematically changed from 1.5 mm to 4.5 mm while holding the most apical active electrode fixed. In Experiment III, the spacing of active electrodes was varied to represent equal tonotopic spacing to equal linear frequency intervals between pairs. Recognition of medial vowels and consonants was measured in three subjects with these custom four-electrode speech processors. RESULTS: In Experiment I, results showed that both vowel and consonant recognition were best when the electrodes were in the most apical locations. In Experiment II, best speech recognition occurred when electrode pairs were separated by 3 to 3.75 mm. In Experiment III, both vowel and consonant recognition scores decreased when the spacing of electrode pairs was changed from equal tonotopic spacing to equal linear frequency intervals. Overall, vowel and consonant recognition were best at the most apical electrode locations and when the spacing of electrodes matched the frequency intervals of the analysis filters. Consonant recognition was relatively robust to alterations in electrode location and spacing. The best vowel scores with four-electrode speech processors were about 10 percentage lower than scores obtained with the full 20-electrode speech processors. However, the best consonant scores with four-electrode speech processors were similar to those obtained with the full 20-electrode speech processors. Information transmission analysis revealed that temporal envelope cues (voicing and manner) were not strongly affected by changes in electrode location and spacing, whereas spectral cues, as represented by vowel recognition and consonantal place of articulation, were strongly affected. Both spectral and temporal phoneme cues were strongly affected by the degree of tonotopic warping, created by altering both the location and spacing of the activated electrodes. CONCLUSION: The cochlear location and spacing of the activated electrodes had a clear effect on phoneme recognition. Temporal cues were less affected by tonotopic shifts or linear tonotopic stretching or shrinking, but were susceptible to nonlinear tonotopic warping. Spectral cues were sensitive to all tonotopic manipulations: shifting, linear stretching, and nonlinear warping. However, the present experiments could not differentiate whether the optimal mapping between analysis frequency bands and stimulation electrodes was determined by the normal acoustic tonotopic pattern or by the pattern learned from experience with the 20-electrode implant. PMID- 10466569 TI - Effects of electrode configuration and frequency allocation on vowel recognition with the Nucleus-22 cochlear implant. AB - OBJECTIVE: This study was conducted to understand vowel recognition in cochlear implants as a function of the cochlear location and separation of the stimulated electrode pairs and as a function of the matching between speech spectral information and the location of the stimulated electrodes. DESIGN: Four-electrode speech processors with a continuous interleaved sampling speech processing strategy were implemented through a custom interface in five subjects implanted with the Nucleus-22 cochlear implant. The temporal envelopes from four broad frequency bands were used to modulate 500 pps, 100 microsec/phase interleaved pulse trains delivered to four electrode pairs. Ten different frequency allocations and five sets of four-electrode configurations were tested. Each frequency allocation represented the same cochlear extent but different cochlear locations based on Greenwood's frequency-to-place formula. Recognition of multi talker medial vowels was measured for each combination of parameters with no period of practice or adjustment. RESULTS: Results showed that recognition of multi-talker vowels was highly dependent on frequency allocation for all electrode configurations. For a given electrode configuration maximum vowel recognition was observed with a specific frequency allocation. When the stimulated electrodes were shifted basally by 3 mm, the frequency allocation that produced the best performance also shifted basally by 3 mm. A similar pattern of vowel recognition was observed as a function of frequency allocation for electrode configurations that had the same apical-most electrode in each pair, regardless of location of the basal-most electrode in the pair. Subjects with different electrode insertion depths had similar trends in vowel recognition for each frequency allocation. CONCLUSIONS: For a given electrode configuration, the best performance was obtained with processors with a specific frequency allocation. In addition, the apical-most member of each electrode pair had a much stronger influence on vowel recognition in electric hearing. Finally, results from this study also suggest that over time, patients with implants can partially adapt to a basal shift in place of stimulation. PMID- 10466570 TI - Distortion product otoacoustic emission test performance for a priori criteria and for multifrequency audiometric standards. AB - OBJECTIVES: 1) To describe distortion product otoacoustic emission (DPOAE) test performance when a priori response criteria are applied to a large set of DPOAE data. 2) To describe DPOAE test performance when multifrequency definitions of auditory function are used. 3) To determine DPOAE test performance when a single decision regarding auditory status is made for an ear, based on DPOAE data from several frequencies. 4) To compare univariate and multivariate test performance when multifrequency gold standard definitions and response criteria are applied to DPOAE data. DESIGN: DPOAE and audiometric data were analyzed from 1267 ears of 806 subjects. These data were evaluated for three different frequency combinations (2, 3, 4 kHz; 2, 3, 4, 6 kHz; 1.5, 2, 3, 4, 6 kHz). DPOAE data were collected for each of the f2 frequencies listed above, using primary levels (L1/L2) of 65/55 dB SPL and a primary ratio (f2/f1) of 1.22. Sensitivity and specificity were evaluated for signal to noise ratios (SNRs) of 3, 6, and 9 dB, which are in common clinical use. In addition, test performance was evaluated using clinical decision theory, following the convention we have used in previous reports on otoacoustic emission test performance. Both univariate and multivariate analyses techniques were applied to the data. In addition to evaluating DPOAE test performance for the case when audiometric and f2 frequency were equal, multifrequency gold standards and multifrequency criterion responses were evaluated. Three new gold standards were used to assess test performance: average pure-tone thresholds, extrema thresholds that took into account both the magnitude of the loss and the number of frequencies at which hearing loss existed, and a combination of the two. These new gold standards were applied to each of the three frequency groups described above. RESULTS: As expected, SNR criteria of 3, 6, and 9 dB never resulted in perfect DPOAE test performance. Even the most stringent of these criteria (9 dB SNR) did not result in a sensitivity of 100%. This result suggests that caution should be exercised in the interpretation of DPOAE test results when these a priori criteria are used clinically. Excellent test performance was achieved when auditory status was classified on the basis of the new gold standards and when either SNR or the output of multivariate logistic regressions (LRs) were used as criterion measures. Invariably, the LR resulted in superior test performance compared with what was achieved by the SNR. For SNR criteria of 3, 6, and 9 dB and (by definition) for the LR, specificity, in general, exceeded 80% and often was greater than 90%. Sensitivity, however, depended on the magnitude of hearing loss. Diagnostic errors, when they occurred, were more common for patients with mild hearing losses (21 to 40 dB HL); sensitivity approached 100% once the hearing loss exceeded 40 dB HL. The largest differences between test performance based on SNR or LR occurred for the ears with mild hearing loss, where the LR resulted in more accurate diagnoses. CONCLUSIONS: It should not be assumed that the use of a priori response criteria, such as SNRs of 3, 6, or 9 dB, will identify all ears with hearing loss. Test performance when multifrequency gold standards are used to define an ear as normal or impaired and when data from multiple f2 frequencies are used to make a diagnosis, resulted in excellent test performance, especially when the LR was used. When predicting auditory status with multifrequency gold standards, the LR resulted in relative operating characteristic curve areas of 0.95 or 0.96. An output from the LR can be selected that results in a specificity of 90% or better. When the loss exceeded 40 dB HL, the same output from the LR resulted in test sensitivity of nearly 100%. These were the best test results that were achieved. (ABSTRACT TRUNCATED) PMID- 10466571 TI - Some computational analyses of the PBK test: effects of frequency and lexical density on spoken word recognition. AB - OBJECTIVE: The Phonetically Balanced Kindergarten (PBK) Test (Haskins, Reference Note 2) has been used for almost 50 yr to assess spoken word recognition performance in children with hearing impairments. The test originally consisted of four lists of 50 words, but only three of the lists (lists 1, 3, and 4) were considered "equivalent" enough to be used clinically with children. Our goal was to determine if the lexical properties of the different PBK lists could explain any differences between the three "equivalent" lists and the fourth PBK list (List 2) that has not been used in clinical testing. DESIGN: Word frequency and lexical neighborhood frequency and density measures were obtained from a computerized database for all of the words on the four lists from the PBK Test as well as the words from a single PB-50 (Egan, 1948) word list. RESULTS: The words in the "easy" PBK list (List 2) were of higher frequency than the words in the three "equivalent" lists. Moreover, the lexical neighborhoods of the words on the "easy" list contained fewer phonetically similar words than the neighborhoods of the words on the other three "equivalent" lists. CONCLUSIONS: It is important for researchers to consider word frequency and lexical neighborhood frequency and density when constructing word lists for testing speech perception. The results of this computational analysis of the PBK Test provide additional support for the proposal that spoken words are recognized "relationally" in the context of other phonetically similar words in the lexicon. Implications of using open-set word recognition tests with children with hearing impairments are discussed with regard to the specific vocabulary and information processing demands of the PBK Test. PMID- 10466572 TI - Reflections on a century of primatology. PMID- 10466573 TI - Proposed chromosomal phylogeny for the South American primates of the Callitrichidae family (Platyrrhini). AB - Cytogenetic and cytotaxonomic studies (G, C, sequential G/C, and NOR banding) were performed on 110 specimens representing the four genera of South American primates of the family Callitrichidae: Cebuella (C. pygmaea), Callithrix, groups argentata (C. argentata, C. emiliae, C. chrysoleuca, C. humeralifera, C. mauesi), and jacchus (C. aurita, C. geoffroyi, C. jacchus, C. kuhli, C. penicillata), Leontopithecus (L. chrysomelas, L. rosalia), and Saguinus (S. midas midas, S. m. niger). Mitotic chromosomes are characterized, and the rearrangements distinguishing the karyotypes of the taxa are inferred from arm homologies. The results were then converted into numerical data and submitted to cladistic analysis. The following conclusions were achieved: 1) Five karyotypic classes were observed, which correspond to the five taxa studied. Differences between them are as follows: a) Cebuella (2n = 44, 10 acrocentrics, A + 32 bi-armed autosomes, bi) and the argentata group (2n = 44, 10A + 32bi) are different from each other due to a reciprocal translocation; b) both can be distinguished from the jacchus group (2n = 46, 14A + 30bi) by a centric fusion/fission rearrangement and a paracentric inversion; c) Leontopithecus (2n = 46, 14A + 30bi) and Saguinus (2n = 46, 14A + 30bi) differ from the jacchus group by a reciprocal translocation and three paracentric inversions; and d) Saguinus is different from the others by one paracentric inversion and pericentric inversions in at least four pairs of acrocentric autosomes. 2) The cladistic analysis separates Cebus (used as an outgroup) from the Callitrichidae groups, which forms a clade. Among the Callitrichidae, marmosets (Cebuella and Callithrix) form a sub-clade, Cebuella and the argentata group being more closely related to each other than both are to the jacchus group. Tamarins (Leontopithecus and Saguinus) are also quite close, so that if one was not derived from the other, they with the marmosets share a common ancestor. Among the tamarins, Leontopithecus is karyotypically closest to the marmosets, specifically to the jacchus group. 3) Based on the chromosome information and considering the possible direction of the evolutionary changes (primitivity or phyletic dwarfism hypothesis, previously advanced by other authors), it was possible to propose the ancestral karyotypes and to develop two alternatives for the origin, differentiation and dispersion of the callitrichid. Both proposals are plausible, but when the geographical distribution is considered, the phyletic dwarfism hypothesis seems to be the most probable. PMID- 10466574 TI - Incorporation of a GnRH agonist, leuprolide acetate, into regimens with exogenous gonadotropins to produce ovarian stimulation and ovulation in the nonpregnant squirrel monkey. AB - This study was designed to measure the effects of variations in the length of pretreatment with a GnRH agonist, leuprolide acetate (LA), on subsequent follicular development and ovulation. The hypothesis was that the duration of LA suppression of pituitary function does not adversely affect ovarian response to standardized ovulation induction protocols in squirrel monkeys. The first phase determined the dose and duration of LA needed to achieve a hypogonadal state. One of two groups received daily subcutaneous injections of 50 microg of LA. The other received a single injection of 175 microg of a depot suspension of LA. Sera were assayed for estradiol (E2) and progesterone (P). E2 and P levels increased 2 to 5-fold with peak levels on days 4 and 7, respectively. Suppression of steroid levels took 10 to 15 days in the LA-treated group. Depot-LA did not effectively suppress steroid production. After suppression, females receiving daily LA received five daily injections of hMG to stimulate follicular development. E2 and P increased in these animals. These results suggest that cycling squirrel monkeys have P-secreting capacity throughout the cycle. This may explain how the squirrel monkey is able to accommodate both a short (4-5 day) luteal phase of their 9 day cycle and implantation from 5 to 7 days after ovulation. A second study compared exogenous follicle stimulating hormone (FSH) to endogenous gonadotropins released as a response to LA in ovulation induction. Steroid production and hCG-induced ovulation were assessed. LA treatment was compared to a standard ovulation induction protocol by using a randomized cross-over measures design. There were no differences in E2 and P levels in response to dosages of either LA or hMG. The ovulatory response following LA treatment was not significantly greater than that using FSH. The number of animals with unovulated, large follicles was greater on the FSH protocol (12/18) compared to the LA protocol (3/18). Thus, a single injection of a depot preparation of LA is sufficient to stimulate follicular development and ovulation when followed by an hCG injection. Based on this observation and the data on unovulated large follicles, it is suggested that the ovary responds more readily to endogenous gonadotropins released by LA than to exogenous FSH. PMID- 10466575 TI - Context and sex differences exist in the acoustic structure of phee calls by newly-paired common marmosets (Callithrix jacchus). AB - Captive common marmosets of all ages robustly produce a "separation" phee call during brief separations from their group. In contrast, a second structural variant, which may function as an intergroup call, is produced in the home cage primarily by the reproductive adults. A previous study found that postpubertal but nonreproductive offspring rarely produce phee calls when in the home cage with the natal group, yet these marmosets call frequently after pairing with an opposite-sex partner. The sudden increase in home cage phee calls may indicate the rapid onset of intergroup calling. Alternatively, marmosets may be producing the separation phee variant as a result of separation from the natal group. The present study investigated whether phee calls produced by recently paired individuals in the home cage were structurally distinguishable from their calls recorded in a separation paradigm. We also tested whether sex differences, known to exist in the calls of mature adults, could be found in calls recorded from younger, nonreproductive animals separated from their natal groups. We analyzed 18 acoustic parameters of phee calls produced in the home cage after pairing and of calls produced during separation both from the natal group and from a new mate. Discriminant function analyses found that home cage calls were clearly discriminable from separation calls (average 91.7% correctly classified), indicating that the rapid increase in home cage phee call production shortly after pairing is not a consequence of separation from the family group. Postpubertal marmosets appear to show a rapid behavioral adjustment to separation from their natal groups. Additionally, sex was clearly discriminable in calls recorded both before and after pairing (average 86.8% correctly classified). Like calls recorded from well-established paired marmosets, phee calls produced by recently paired, postpubertal marmosets are discriminable by context and sex. PMID- 10466576 TI - Topographical localization of lipofuscin pigment in the brain of the aged fat tailed dwarf lemur (Cheirogaleus medius) and grey lesser mouse lemur (Microcebus murinus): comparison to iron localization. AB - The present study was undertaken to explore the distribution of lipofuscin in the brain of cheirogaleids by autofluorescence and compare it to other studies of iron distribution. Aged dwarf (Cheirogaleus medius) and mouse (Microcebus murinus) lemurs provide a reliable model for the study of normal and pathological cerebral aging. Accumulation of lipofuscin, an age pigment derived by lipid peroxidation, constitutes the most reliable cytological change correlated with neuronal aging. Brain sections of four aged (8-15 year old) and 3 young (2-3 year old) animals were examined. Lipofuscin accumulation was observed in the aged animals but not in the young ones. Affected regions include the hippocampus (granular and pyramidal cells), where no iron accumulation was observed, the olfactory nucleus and the olfactory bulb (mitral cells), the basal forebrain, the hypothalamus, the cerebellum (Purkinje cells), the neocortex (essentially in the pyramidal cells), and the brainstem. Even though iron is known to catalyse lipid oxidation, our data indicate that iron deposits and lipofuscin accumulation are not coincident. Different biochemical and morphological cellular compartments might be involved in iron and lipofuscin deposition. The nonuniform distribution of lipofuscin indicates that brain structures are not equally sensitive to the factors causing lipofuscin accumulation. The small size, the rapid maturity, and the relatively short life expectancy of the cheirogaleids make them a good model system in which to investigate the mechanisms of lipofuscinogenesis in primates. PMID- 10466577 TI - Can puzzle feeders be used as cognitive screening instruments? Differential performance of young and aged female monkeys on a puzzle feeder task. AB - Conventional cognitive testing of monkeys is time-consuming and involves single caging and food or water deprivation. Here we report a novel test of global cognitive performance that can be completed in a short time period without food/water or social restrictions. Nine mazes of increasing difficulty were developed using a standard puzzle feeder, and the maze-solving performance of ten young and five aged female cynomolgus monkeys (Macaca fascicularis) was tested. The young monkeys solved maze configurations at higher levels of difficulty and solved the first level of difficulty more quickly than aged monkeys. This task discriminated performance by age in nonhuman primates as do more conventional forms of cognitive testing and indicates that this task may be a quick and easy assessment of global cognitive function. PMID- 10466579 TI - Evaluation of the effects of Gd complexes used as magnetic resonance imaging contrast agents, on superoxide dismutase: comparison of two methods. AB - Investigations are currently being made into the safety of gadolinium complex contrast agents used in Magnetic Resonance Imaging. Their hyperosmolality or potential Gd3+ release is evoked as a cause of various anaphylactoid reactions to be observed in humans after intravenous injection. An estimation has already been made of their effects on the liberation of reactive oxygen intermediates by neutrophils. The purpose of this study was to find a suitable method to measure SOD activity in the presence of these hyperosmolar solutions, and to evaluate their action on this activity. Two techniques were compared to measure this activity. Results and statistical analysis showed that pyrogallol autoxidation was greatly affected by solution osmolalities, whereas ferricytochrome C reduction was not. Gadopentetate dimeglumine and gadoterate meglumine seemed to activate Cu, Zn SOD in vitro, but did not exhibit any SOD-like activity. Gadodiamide did not interfere with this system of detoxication. PMID- 10466578 TI - Rat macrophage inflammatory protein-1alpha, a CC chemokine, acts as a neutrophil chemoattractant in vitro and in vivo. AB - Recombinant rat macrophage inflammatory protein-1alpha (rMIP-1alpha) at a concentration of 3x10(-8) M had strong neutrophil chemotactic activity, though the potency of rMIP-1alpha was less than that of cytokine-induced neutrophil chemoattractant (CINC)-1 at lower concentrations. In addition, rMIP-1alpha induced neutrophil chemotaxis in vivo when rMIP-1alpha was injected into the preformed air-pouch on the back of rats. The adhesion of rMIP-1alpha-treated neutrophils to fibrinogen significantly increased, reaching a maximum adhesion at 10(-8) M. Stimulation of neutrophils with rMIP-1alpha induced a transient increase in intracellular free [Ca2+] dose-dependently. rMIP-1alpha still induced an increase in the intracellular [Ca2+] of rat neutrophils stimulated first with CINC-1, CINC-3 or C5a, suggesting that rat neutrophils have a specific receptor for rMIP-1alpha. Supporting these findings, an additive increase in chemotactic potency was found when both rMIP-1alpha and CINC-were added to the lower wells of Boyden chamber in vitro. In addition, high levels of rMIP-1alpha were detected in the inflammatory site of air-pouch/carrageenan-induced inflammation in rats. Our results suggest that rMIP-1alpha acts as a neutrophil chemoattractant and, together with CINCs, plays an important role in infiltration of neutrophils into inflammatory sites in rats. PMID- 10466580 TI - Early bronchial hyperresponsiveness following injection of sephadex beads in the guinea pig: involvement of platelet activating factor and thromboxane A2. AB - The effects of an intravenous injection (i.v.) of Sephadex beads (20 mg kg(-1)) were examined on bronchial responsiveness to ACh (1-200 microg kg(-1) i.v.) as well as on cell accumulation in guinea-pig lung. Bronchial hyperreactivity to ACh, measured as increase in pulmonary insufflation pressure (PIP), was observed 3 h following the i.v. injection of Sephadex beads. However, no significant increase in bronchial reactivity to ACh was measured at 6 and 12 h following Sephadex injection. A second later increase in bronchial hyperresponsiveness was observed at 24 h. Bronchoalveolar lavage performed at 3 h following Sephadex treatment showed that there was no significant increase in total or differential cell number. At 6 h and 12 h, a significant increase in total cell counts was observed. At 24 h, a greater than 5-fold increase in cell number was observed and was related to a marked eosinophil, neutrophil and macrophage infiltration. A platelet-activating factor (PAF) antagonist, CV-3988 (10 mg kg(-1) i.v.), and a thromboxane A2 (TxA2) antagonist, L655,240 (10 mg kg(-1) i.v.), significantly attenuated the Sephadex-induced bronchial hyperresponsiveness to ACh observed at 3 h. The results show that an i.v. injection of Sephadex beads in guinea pigs can induce an early bronchial hyperresponsiveness to ACh that is mediated by the release of both PAF and TxA2 and is independent of airway cell infiltration. PMID- 10466581 TI - Protection by vitamin B2 against oxidant-mediated acute lung injury. AB - The effect of vitamin B2 (riboflavin) on oxidant-mediated acute lung injury has been examined in three different rat models. Pulmonary injury was induced by intravenous injection of cobra venom factor (CVF), by the intrapulmonary deposition of IgG immune complexes, or by hind limb ischemia-reperfusion. In each of the three models, injury was characterized by increases in vascular permeability (leakage of 125I-labeled bovine serum albumin), alveolar hemorrhage (extravasation of 51Cr-labeled rat erythrocytes), and neutrophil accumulation (myeloperoxidase activity). Intraperitoneal administration of riboflavin at a dose of 6 micromoles/kg body weight reduced vascular leakage by 56% in the CVF model, by 31% in the immune complex model, and by 53% in the lung injury model following ischemia-reperfusion of the hind limbs. Similar treatment reduced hemorrhage by 76%, 51%, and 70% in the three models of lung injury. In the CVF model, riboflavin was also shown to decrease products of lipid peroxidation (conjugated dienes) in lungs (by 45%) and in plasma (by 74%). Neutrophil accumulation in the lungs was not influenced by riboflavin administration in any of the three models. The studies demonstrate that riboflavin can mount a significant protection against oxidant-mediated inflammatory organ injury. PMID- 10466582 TI - Protective effect of anti-P-selectin monoclonal antibody in lipopolysaccharide induced lung hemorrhage. AB - Excessive leukocyte accumulation is involved in the pathogenesis of the sepsis induced acute lung injury. Selectins are essential to the interaction between leukocytes and endothelial cells. In this report, we investigated the role of selectins in the severe lung injury induced by lipopolysaccharide (LPS). Significant lung hemorrhage was observed 24 h after the intravenous administration of LPS (1 mg/kg). First, we evaluated the effect of sialyl Lewis X oligosaccharide (SLeX-OS), a derivative of sialyl Lewis X which is one of the ligands for E-, P- and L-selectins. The treatment with SLeX-OS (26.5 mg/kg iv bolus + 19.8 mg/kg iv infusion) resulted in a decrease of lung hemorrhage by 49.5% (P<0.05 versus the control group). Second, we tested the effect of anti-P selectin monoclonal antibody (MAb), PB 1.3, to investigate the role of P selectin. The bolus administration of PB1.3 at a dose of 5 mg/kg attenuated the lung hemorrhage by 74.6% (P<0.05 versus the control group). In addition, we also detected an increase of soluble P-selectin in plasma 24 h after the injection of LPS. These results suggest that P-selectin has a substantial role in the pathogenesis of the lung injury induced by LPS. PMID- 10466583 TI - Expression of FCepsilonII/CD23 on human neutrophils isolated from rheumatoid arthritis patients. AB - CD23, the low affinity receptor for IgE, is a 45 kilodalton molecule belonging to the C-type lectin family, some members of which have been identified as adhesion molecules. Since it has been described upregulated in different cells in chronic inflammatory diseases and in rheumatoid arthritis in particular, where neutrophils are directly involved in tissue damage, our interest, in this work, has been focused on the expression and regulation of this antigen on neutrophil membrane. We studied 22 patients suffering from rheumatoid arthritis and 22 healthy control subjects. CD23 expression on neutrophil membrane was analyzed by immunofluorescence. Neutrophils of 9 out of 22 patients expressed CD23 molecules, neutrophils of 11 out of 22 patients expressed CD23 only after 24 h of incubation in RPMI; only 2 out of 22 patients did not express the CD23 antigen on neutrophil membrane either after isolation or after a 24 h incubation. On the contrary neutrophils isolated from healthy subjects did not express CD23 molecules upon isolation. Only in 7/22 control subjects neutrophils resulted positive after 24 h of incubation in RPMI. Moreover, we found that in our experimental conditions the presence of IFN-g or GM-CSF alone or in combination with IL-4 inhibited CD23 expression during the 24 h incubation. Our results show that there is a strong association between neutrophil ability to express CD23 and rheumatoid arthritis, and that such expression may be regulated by GM-CSF, IFN-gamma and IL-4. PMID- 10466584 TI - Prostaglandin E2 downregulates interferon-gamma-induced intercellular adhesion molecule-1 expression via EP2 receptors in human gingival fibroblasts. AB - In the present study, the effect of prostaglandin E2 (PGE2) on intercellular adhesion molecule-1 (ICAM-1) expression in interferon-gamma (IFN-gamma) stimulated human gingival fibroblasts (HGF) was investigated. Addition of PGE2 to HGF inhibited ICAM-1 expression elicited by IFN-gamma. As PGE2 elevated intercellular cyclic AMP (cAMP) levels in HGF in a dose-dependent fashion, the effect of dibutyryl cAMP and 8-bromo-cAMP, cAMP analogues, on IFN-gamma-elicited ICAM-1 expression was examined. Both the agents downregulated ICAM-1 expression in IFN-gamma-stimulated HGF. Next, we examined which subtype(s) of the four PGE2 receptor subtypes (EP1, EP2, EP3 and EP4) modulated the ICAM-1 expression elicited by IFN-gamma, using subtype-specific agonists or antagonists. An EP2/EP4 agonist, 11-deoxy-PGE1, attenuated IFN-gamma-elicited ICAM-1 expression in a concentration-dependent manner. A specific EP4 antagonist, AH-23848B, showed no effect on inhibition of IFN-gamma-elicited ICAM-1 expression by PGE2 and 11-deoxy PGE1. Butaprost, an EP2-selective agonist, mimicked inhibition of IFN-gamma elicited ICAM-1 expression by 11-deoxy-PGE1. An EP3 agonist, ONO-AP-324, was inert with respect to IFN-gamma-elicited ICAM-1 expression. Sulprostone, an EP1/EP3 agonist, showed stimulatory effect on ICAM-1 expression elicited by IFN gamma. From these results, we suggest that PGE2 downregulates IFN-gamma-induced ICAM-1 expression in HGF, primarily via EP2 receptors by cAMP-dependent signaling pathways. PMID- 10466585 TI - Drusen in age-related macular degeneration: pathogenesis, natural course, and laser photocoagulation-induced regression. AB - Drusen are subretinal pigment epithelial deposits that are characteristic of but not uniquely associated with age-related macular degeneration (AMD). Age-related macular degeneration is associated with two types of drusen that have different clinical appearances and different prognoses. Hard drusen appear as small, punctate, yellow nodules and can precede the development of atrophic AMD. Areolar atrophy of the retinal pigment epithelium (RPE), choriocapillaris, and outer retina develop as the drusen disappear, but drusen can regress without evidence of atrophy. Soft drusen appear as large (usually larger than 63 microm in diameter), pale yellow or grayish-white, dome-shaped elevations that can resemble localized serous RPE detachments. They tend to precede the development of clinically evident RPE detachments and choroidal neovascularization. Drusen characteristics correlated with progression to exudative maculopathy include drusen number (five or more), drusen size (larger than 63 microm in diameter), and confluence of drusen. Focal hyperpigmentation in the macula and systemic hypertension also are associated with an increased risk of developing choroidal new vessels (CNVs). Large drusen are usually a sign of diffuse thickening of Bruch's membrane with basal linear deposit, a vesicular material that probably arises from the RPE, constitutes a diffusion barrier to water-soluble constituents in the plasma, results in lipidization of Bruch's membrane, and creates a potential cleavage plane between the RPE basement membrane and the inner collagenous layer of Bruch's membrane through which CNVs can grow. Disappearance of drusen spontaneously and in areas adjacent to laser photocoagulation scars was first noted by Gass (Gass JD: Arch Ophthalmol 90:206 217, 1973; Trans Am Acad Ophthalmol Otolaryngol 75:580-608, 1971). Subsequent reports have confirmed these observations. Photocoagulation-induced drusen regression might prevent patients with drusen from developing exudative maculopathy. The mechanism for spontaneous drusen regression probably involves RPE atrophy. The mechanism for photocoagulation-induced drusen regression is unknown. If photocoagulation-induced drusen regression is anatomically similar to atrophy-associated drusen regression, then the former will be associated with dissolution of basal linear deposit and a residuum of basal laminar deposit. Sarks and coworkers (Sarks JP, Sarks SH, Killingsworth MC: Eye 11:515-522, 1997) proposed that this in turn will eliminate the potential cleavage plane between the RPE basement membrane and inner collagenous layer of Bruch's membrane through which CNVs grow, thus retarding the growth of CNVs. PMID- 10466586 TI - The placebo effect. AB - Placebos have been traditionally regarded as deceptive therapies and have not been understood in the broader context of social symbols and of interpersonal factors that surround the healing process itself. Although the power of inert substances to heal is well recognized, the placebo effect also influences the outcome of conventional therapies. The role of the placebo in modern medicine is poorly defined because of a lack of a common understanding of what the placebo effect is and because of the negative connotions associated with its use. The response rate to placebo varies by illness. The natural course of disease and patient or physician bias can be misinterpreted as a placebo response. In research, the placebo effect is therapeutic noise to be removed by placebo controlled trials. Few studies are designed to measure the placebo response rate directly. Placebos are a reminder of how little is known about mind-body interaction. The placebo effect may be one of the most versatile and underused therapeutic tools at the disposal of physicians. PMID- 10466587 TI - Double vision in a child. AB - A 10-year-old boy presented with a complete left oculomotor cranial nerve palsy. Diagnostic evaluation, including neuroimaging and cerebral angiography revealed a small intracranial aneurysm compressing the third nerve. Neurosurgical clipping of the aneurysm produced resolution of the third nerve palsy. The rarity of this presentation in a young patient is discussed, along with the importance of rapid diagnosis and treatment. PMID- 10466588 TI - The electroretinogram in diabetic retinopathy. AB - Electroretinography (ERG) is an objective method of evaluating retinal function. Since its introduction to clinical practice in the 1940s, it has become a useful and routine diagnostic clinical tool in ophthalmology. This review summarizes the role of ERG as a clinical technique for evaluating the progression of diabetic retinopathy and as a research tool for increasing our understanding of the pathophysiology of diabetic retinopathy. Most studies show unequivocally that the different types of ERG tests detect local abnormalities or widespread pathology, even in very early stages of the disease. It seems plausible that measurements from ERG recordings, particularly the oscillatory potentials, may be useful for predicting progression from nonproliferative to the more sight-threatening stages -preproliferative or proliferative--of diabetic retinopathy. Some recent work implies that the ERG can also be a useful diagnostic method for discriminating between eyes with diabetic retinopathy and those without the condition. PMID- 10466589 TI - Telemedicine and ophthalmology. AB - Technology to create and move multimedia medical information is creating alternatives to physically transporting patients and health care professionals. Teletechnology is a physician extender, both supplementing and reinventing traditional health care delivery systems. The potential of comprehensive teleophthalmology is compelling. This article provides a historical perspective of telemedicine, describes various teleophthalmology systems, and surveys teleophthalmology in clinical consultation, research, and education programs around the world. It also discusses basic issues in applying teleophthalmology to public health. PMID- 10466590 TI - "Eyes" on the thrones: imperial ophthalmologic nicknames. AB - Several Greek and Byzantine sovereigns are known in history by nicknames that are of ophthalmologic origin; the sobriquets derive from characteristics of their eyes or their actions in relation to the eyes. The first was Antigonos I Monophthalmus (the One-eyed), who was the most eminent successor of Alexander the Great and Sovereign of Eastern Mediterranean Asia. He obtained his nickname at an early age when he lost one eye fighting at the seige of Perinthos, as a general of King Philip of Macedon (father of Alexander the Great). Several Byzantine emperors also have a number of similar nicknames, such as Anastasius I Dicoros (with Heterochromia Iridis), Alexius V Ducas Murtzuphlus (with Scowling Eyebrows), and Andronicus I Comnenus Misophaes (Hater of Sunlight, since he blinded a great number of his opponents). To these must be added the Byzantine Empress Zoe Carvounopsina (with Coal-black Eyes). PMID- 10466591 TI - Kimura disease of the orbit and ocular adnexa. AB - Kimura disease (KD) is a distinct clinicopathologic entity that has been the subject of considerable confusion and debate. Although common in Asia, KD rarely occurs in non-Asian patients. Kimura disease shares both clinical and histopathologic features with angiolymphoid hyperplasia with eosinophilia (ALHE). Because of this overlap and the rarity of KD in Europe and the United States, KD and ALHE have been used synonymously in the Western medical literature, as they were thought to represent variations of the same disease. Some pathologic reports have called for distinguishing KD and ALHE as two separate entities, based on their histologic features. Kimura disease occurs most commonly in the head and neck region and has been described in the orbit, eyelids, and lacrimal gland more frequently than ALHE. Because both diseases can cause proptosis, lid swelling, ocular dysmotility, or a palpable mass, they should be considered in the differential diagnosis of orbital lesions occurring in adults. We report two cases of KD involving the orbit and ocular adnexa, and review additional cases reported in the literature. The ophthalmic literature does not clearly reflect the current understanding that KD and ALHE are best considered two separate clinicopathologic entities. PMID- 10466592 TI - Reducing aniseikonia. PMID- 10466593 TI - Grading conjunctivochalasis. PMID- 10466595 TI - A population-based study of congenital diaphragmatic hernia: impact of associated anomalies and preoperative blood gases on survival. AB - BACKGROUND/PURPOSE: Although neonatal care has improved over the past 20 years, mortality rate with congenital diaphragmatic hernia (CDH) remains 50% to 60%, possibly reflecting differences in management or selection biases. The authors determined the incidence, outcome, effect of coexisting anomalies, and prognostic indicators for neonates with CDH in a single inborn population older than 13 years. METHODS: Forty-three neonates with CDH, those symptomatic within the first 6 hours of life, were identified using a validated neonatal database and diagnosis coding data from medical records among 180,643 live inborn neonates delivered at Parkland Memorial Hospital between 1983 and 1995. Charts were reviewed for prenatal history, demographic variables, presence of coexisting malformations, preoperative arterial blood gases, surgical findings, and outcome. Survival to hospital discharge was the primary outcome variable. RESULTS: The incidence of CDH was 1 in 4,200 live births; overall survival rate was 51%. Thirty-two (74%) neonates underwent surgical repair, often at less than 8 hours of life; postoperative mortality rate was 31%. Eighteen (42%) had coexisting major anomalies or chromosomal abnormalities. Eighty percent of neonates with isolated CDH survived, whereas 89% with CDH and associated defects died. Nonsurvivors had lower birth weights and Apgar scores, were more acidotic, and had more severe respiratory compromise. When best preoperative pH was > or = 7.25 or PaCO2 < or = 50 mm Hg, 80% of neonates survived. CONCLUSION: In this inborn population-based review of neonatal CDH between 1983 and 1995, the best predictors of survival were the presence or absence of other anomalies and the best preoperative PaCO2 and pH. PMID- 10466594 TI - Effects of dexamethasone, growth factors, and tracheal ligation on the development of nitrofen-exposed hypoplastic murine fetal lungs in organ culture. AB - BACKGROUND/PURPOSE: The addition of growth factors EGF (epidermal growth factor) plus TGFbeta1 (transforming growth factor beta1; E + T) or dexamethasone (DEX) to normal murine fetal lungs in culture enhances lung development. In addition, ligation of the airway in lungs in organ culture, enhances lung development. Nitrofen (2,4-dichlorophenyl-p-nitrophenylether) administration to pregnant mice results in pulmonary hypoplasia in the offspring with many similarities to human hypoplastic lung conditions. This study investigates the effects of growth factors, dexamethasone, and airway ligation on the development of hypoplastic fetal murine lungs in whole-organ culture. We hypothesized that E+T, DEX, or airway ligation will enhance the development and maturation of hypoplastic murine fetal lungs in vitro. METHODS: Time-dated pregnant CD-1 mice were given nitrofen, 25 mg, intragastrically at gestational day (Gd) 8. The dams were killed on Gd 14, and the fetuses were removed. The hypoplastic fetal lungs were excised, and the tracheae were transected. The lungs were cultured in serum-free BGJb media in the presence or absence of E+T (10 ng/mL + 2 ng/mL, respectively) or DEX (10 nmol/L). Some lungs were cultured for 7 days with the tracheae ligated. RESULTS: Gross morphology under a dissecting stereomicroscope showed that the lungs were larger after E+T, DEX, or tracheal ligation. Histologically, the untreated lungs had progressed from the pseudoglandular stage to a canalicular-like stage with poorly differentiated airways. The E+T-treated lungs had better developed airway branching and small acini; however, thick mesenchyme persisted. The ligated lungs had well-developed airway branching and acinar structures. After DEX treatment the lungs were most developed with very well defined airway branching and expanded acinar structures; however, there was no secondary septation. Ultrastructurally, the hypoplastic lungs at Gd 14 and after 7 days in culture had no glycogen in their epithelial cells, no defined acinar formation, and had damaged mitochondria. The E+T-treated or tracheally ligated lungs had abundant type II cells, secreted lamellar bodies (LBs), and showed infrequent tubular myelin. Mitochondrial damage was noted in these lungs as in the untreated lungs. DEX-treated hypoplastic lungs showed large acini. The acinar walls were thick; however, they had type II cells with abundant LBs and intact mitochondria. The airways were noted to have differentiated cell types. Surfactant secretions in acinar spaces showed tubular myelin structures. CONCLUSIONS: E+T, tracheal ligation, or DEX accelerates lung development and maturation of hypoplastic fetal murine lungs compared with untreated controls. DEX had a greater effect with special reference to repair of mitochondrial damage. DEX not only accelerated lung development, but it may have reversed some of the effects nitrofen. PMID- 10466596 TI - Cardiovascular malformations in experimental congenital diaphragmatic hernia. AB - BACKGROUND/PURPOSE: Newborns with congenital diaphragmatic hernia (CDH) frequently have associated anomalies that have a major impact on survival rate independent of pulmonary hypoplasia and pulmonary hypertension. Cardiovascular malformations (CVM) represent a major group of lethal extrapulmonary abnormalities that often assume greatest prognostic significance in most CDH studies. Animal models resembling human CDH may aid knowledge of the basic embryology that leads to the coexpression of CDH and CVM. This study, therefore, analyzed the incidence and spectrum of CVM in fetal rats with CDH. METHODS: Left sided CDH (LCDH) was induced in fetal rats by the maternal administration of 100 mg of nitrofen by gavage on day 9.5 gestation (term, day 22). Control animals received olive oil (OO) and were used for comparative analysis. Fetal rats were harvested by cesarean section on day 21.5 or day 22, histologically processed and examined for CVM. RESULTS: A significant number of CVM were observed in 15 of 60 (25%) LCDH rats compared with 4 of 60 (6.7%) nitrofen non-CDH rats (P = .01). The spectrum of abnormalities in CDH included ventricular septal (VSD) defects (n = 6), vascular rings (n = 4), anomalous subclavian arteries (n = 3), atrioventricular septal defects (n = 1) and Fallot's tetralogy (n = 1). A VSD (n = 1), double-outlet right ventricle VSD (n = 1) and Fallot's tetralogy (n = 2) were noted in nitrofen non-CDH rats. Control (OO) fetal rats (n = 60) displayed no malformations. CONCLUSIONS: These results confirm a significant incidence and spectrum of CVM in a teratogenic CDH model similar to that seen in humans with CDH. The findings of this study reinforce the validity of the nitrofen model as a research tool to uncover the genetic and molecular mechanisms responsible for the genesis of CDH and allied malformations. PMID- 10466597 TI - Exogenous calcitonin gene-related peptide can change the direction of gubernacular migration in the mutant trans-scrotal rat. AB - PURPOSE: The mutant trans-scrotal (TS) rat shows unilateral or bilateral suprainguinal testes in more than 70% of males. Calcitonin gene-related peptide (CGRP) has been proposed as the neurotransmitter released from the genitofemoral nerve (GFN), which controls gubernacular migration to the bottom of the scrotum during inguinoscrotal descent. Results of previous studies in this rat suggest a down-regulation of CGRP receptors in gubernaculum occurring caused by excess release of the neuropeptide from the GFN. The aim of this study was to test the hypothesis that division of the GFN in neonatal TS rats, potentially allowing the gubernaculum to become sensitive to CGRP, followed by exogenous CGRP injections, would change the direction of gubernacular migration and the final position of the testis. METHODS: Four study groups were used: group 1 (n = 43), sham operation, in which the peritoneal cavity was opened and the left genitofemoral nerve was exposed but not divided, and oil injections into left hemiscrotum; group 2 (n = 70), division of left GFN and CGRP injections into left hemiscrotum; group 3 (n = 36), sham operation and CGRP injections into left hemiscrotum; group 4 (n = 30), division of left GFN and oil injections into left hemiscrotum. RESULTS: In group 2 (GFN division and CGRP injection), 18 testes were located in a position not previously described in this model. In 16 (23%) rats, the testis was located at the entrance of the internal inguinal ring with the gubernaculum directed down toward the scrotum. This contrasts with the normal position of the testis in the superficial inguinal pouch, where the testis is located superficial to the external oblique muscle, with the gubernaculum directed craniolaterally. In two (3%) rats, there was incomplete descent of the testis. In group 3 (sham operation and CGRP injection), two (6%) testes were located at the internal ring. The remaining testes in the above groups and all of the testes in groups 1 and 4 were found in either the superficial inguinal pouch, completely descended, or adherent to scar tissue. CONCLUSIONS: These findings suggest that division of the GFN in neonatal TS rats followed by CGRP injections into the scrotum can change the direction of gubernacular migration in the TS rat. The effectiveness of this experimental model is limited by the inability to accurately localize injected CGRP into the scrotum. PMID- 10466598 TI - Laparoscopic diagnosis of contralateral patent processus vaginalis in children under 1 year of age with unilateral inguinal hernia: comparison with herniography. AB - PURPOSE: The effectiveness of laparoscopic diagnosis of contralateral patent processus vaginalis (CPPV) in children with unilateral inguinal hernia was evaluated. METHODS: Ninety-three consecutive children under the age of 1 year were operated on for a unilateral inguinal hernia. A contralateral CPPV was diagnosed by laparoscopy via the inguinal hernia sac before ligation. The laparoscopy results of this technique were correlated with those of herniography or inguinal exploration. RESULTS: Laparoscopy was performed on 88 patients; sensitivity was 71% and specificity 89%. The only complication arising from the procedure was wound infection in two patients. CONCLUSION: This method is a simple, safe, and accurate procedure for selecting children for contralateral surgical exploration. PMID- 10466599 TI - Long-segment congenital tracheal stenosis: treatment by slide-tracheoplasty. AB - BACKGROUND/PURPOSE: The prognosis of long-segment funnel-shaped congenital tracheal stenosis improved with surgical management by enlargement-tracheoplasty with a pericardial patch. However, the final outcome is not always satisfying mainly because of the complications related to the pericardial graft and because of misdiagnosed associated cardiovascular malformations. The objective of the report is to show a further improvement of the prognosis by a thorough preoperative planning and the use of a slide-tracheoplasty for the cure of the stenosis, to avoid the major drawbacks of enlargement tracheoplasties. METHODS: Two children, 7 months and 3 years old, underwent a bronchoesophagoscopy, spiral computed tomography, an echocardiogram, and a heart catheterization. The tracheal stenosis and the associated severe cardiovascular malformations were cured during a single operative session under cardiopulmonary bypass; the tracheal stenosis was corrected by a slide-tracheoplasty. RESULTS: The postoperative period was remarkably uneventful and the recovery extremely quick (hospital stay of 18 and 17 days). The midterm results are excellent, and the tracheal growth is not impaired (follow-up of 3 1/2 and 3 years). CONCLUSIONS: According to the authors' experience and to the literature, the slide-tracheoplasty seems to be the most efficient surgical procedure for correction of long-segment funnel-shaped congenital tracheal stenosis. But the success of the treatment depends also on a complete preoperative diagnosis allowing a 1-stage surgical treatment of all associated thoracic malformations. PMID- 10466600 TI - The effects of early and delayed immunosuppression in experimental tracheal transplantation with omentopexy. AB - BACKGROUND/PURPOSE: Best results in experimental tracheal allotransplantation are obtained when metachronous revascularization by omentopexy and immunosuppression are used. Nevertheless, this method of revascularization implies in a 4-day period of ischemia to the graft. The aim of this study was to assess the influence of the 4-day period of ischemia on host sensitization as well as the effect of early or delayed immunosuppression on the outcome of the grafts. METHODS: Thirty rabbits were submitted to tracheal allotransplantation and divided according to position of the graft (orthotopic or heterotopic transplants) and the initiation of immunosuppression (early or delayed). The quality of the revascularization was evaluated by the identification of Indian ink, perfused through the abdominal aorta, inside the submucosal vessels. The outcome of the grafts was evaluated by histological analysis according to a semiquantitative scale of alterations. RESULTS: Grafts were better revascularized in heterotopic position. Grafts with late immunosuppression presented good outcome only when heterotopically positioned. No significant differences were observed in grafts placed heterotopically or orthotopically when immunosuppression was initiated early after the transplant. CONCLUSIONS: Transient ischemia produced by metachronous revascularization is not the single factor responsible for the histological alterations observed in tracheal allografts. These alterations probably also are produced by the activation of immune responses. This activation is more intense in more ischemic grafts, but can be suppressed by early administration of immunosuppression. PMID- 10466601 TI - Eight years' experience with foreign-body aspiration in children: what is really important for a timely diagnosis? AB - BACKGROUND/PURPOSE: Aspiration of foreign bodies remains a major cause of morbidity and mortality in childhood. The aim of this study was to evaluate the predictive diagnostic value of clinical signs and symptoms, the history, and the radiology to perform early diagnosis and therapy. METHODS: From January 1990 to March 1998, 87 children were admitted to the Pediatric Surgery Department of Universita di Padova because of suspected foreign body aspiration. Sensitivity and specificity of the considered diagnostic tools were evaluated. RESULTS: Neither clinical signs and symptoms nor radiology have sufficient diagnostic sensitivity, and especially specificity, on which to rely for the diagnosis. Only the choking crisis, when present in the history, has good sensitivity and specificity (respectively, 96% and 76% in this series). CONCLUSIONS: A choking crisis in the child's history should alert physicians to the possibility of a foreign body aspiration. In the present series, complications always were related to the diagnostic delay. PMID- 10466602 TI - Gastric emptying after fundoplication is dependent on changes in gastric volume and compliance. AB - BACKGROUND/PURPOSE: Nissen gastroesophageal fundoplication (GEF) increases gastric emptying (GE); however, the duration and the mechanisms for this improvement in GE remain unclear. The aim of this study was to evaluate the effects of a GEF on GE of a mixed meal, and to determine the correlation between GE and changes in intragastric pressure (IGP) and compliance. METHODS: Using a radiolabeled mixed meal, GE was measured preoperatively 15 and 30 days after operation in 24 Sprague-Dawley rats divided into SHAM and GEF groups. Results were expressed as percent gastric retention at 90 minutes (GRg90), and time to evacuate 50% of the isotope meal (T1/2). Changes in IGP and compliance were determined at the same time-points using a different set of 20 rats. RESULTS: Fifteen days after surgery, GR90 and T1/2 in the GEF group were reduced significantly when compared with preoperative values but returned to near preoperative values 30 days postoperation. In contrast, rats from the SHAM group showed no change in GR90 and T1/2 at 15 days and 30 days postoperation. Immediately after GEF, maximal distension of the stomach resulted in pressures 65% higher than those recorded before operation (20.2 v 11.7 mm Hg; P< .05), which persisted on the 15th postoperative day (17.7 v 10.7 mm Hg; P<.05). On the 30th postoperative day, however, there was no difference in the IGP between rats undergoing GEF compared with those undergoing a SHAM operation (11.7 v 12.0 mm Hg; P < .05). Similarly, mean gastric compliance decreased significantly immediately after and 15 days after GEF, but returned to preoperative levels 30 days after the operation. CONCLUSIONS: In a rat model, GEF produces a transitory increase in GE, which is related to a simultaneous decrease in gastric volume and compliance. However, 30 days after GEF, associated with an elevated IGP, gastric volume increases and GE returns to preoperative levels. PMID- 10466603 TI - Laparoscopic cholecystectomy in children: lessons learned from the first 100 patients. AB - BACKGROUND/PURPOSE: Laparoscopic cholecystectomy is a very common operation in adults but is relatively infrequently required in children. A retrospective review of 100 consecutive infants and children undergoing laparoscopic cholecystectomies from 1990 to 1998 was performed to see what lessons have been learned from this relatively large population of pediatric patients. RESULTS: The patients ranged in age from 25 to 230 months, with a mean of 105 months. Only 19 patients had hemolytic disease as the etiology for their cholelithiasis. Two patients had biliary dyskinesia. Seventy-eight patients underwent an elective operation. Twenty-two children required urgent hospitalization because of complications from their cholelithiasis: acute cholecystitis (n = 7), jaundice and pain (n = 6), gallstone pancreatitis (n = 5), acute biliary colic (n = 4). All 6 patients who presented with jaundice underwent endoscopic retrograde cholangiopancreatography (ERCP) before their laparoscopic cholecystectomy. Two patients required laparoscopic choledochal exploration. The operating time and postoperative hospitalization were significantly longer (P = .0005) in the complicated group when compared with the elective patients. No significant complications such as the need for reoperation, injury to the choledocuhus or to other viscera, bile leak, or retained choledocholithiasis occurred. CONCLUSIONS: Laparoscopic cholecystectomy is a safe, effective procedure in children for removal of the gallbladder. The exact role of routine cholangiography and ERCP remains unclear. PMID- 10466604 TI - Ontogeny of interstitial cells of Cajal in the human intestine. AB - BACKGROUND/PURPOSE: Interstitial cells of Cajal (ICC) recently have been identified as intestinal pacemaker cells. Abnormalities in ICC are increasingly recognized in a number of neonatal disorders such as infantile hypertrophic pyloric stenosis, Hirschsprung's disease, and transient intestinal pseudo obstruction. The aim of this study was to determine the fetal and postnatal differentiation and development of ICC in the human gastrointestinal tract to aid interpretation of pathological specimens. METHODS: Specimens of human gastrointestinal tract from (1) fetuses (9 to 17 weeks' gestation; n = 12), (2) premature and full-term neonates with non-gut motility-related disorders, (age 26 to 59 weeks' gestation; n = 13), and (3) children (age 4 months to 13 years; n = 7) were immunohistochemically stained with antibodies to c-kit(a marker for ICC) and protein gene product 9.5 (PGP9.5, a marker for neural tissue). RESULTS: (1) C kit-positive ICC were present throughout the gut in all specimens including those from the earliest gestational ages. C-kit and PGP9.5 immunoreactivities were present in different cell populations. (2) The distribution of ICC varied with gestational age and with region of the gut. (3) Maturation of ICC networks continues postnatally in a region-specific manner. CONCLUSIONS: ICC are present from an early stage in human gut development. Interpretation of apparent abnormalities in ICC distribution as being of pathological significance should be tempered by the knowledge that ICC networks continue to develop postnatally and that ICC development varies throughout the gut. PMID- 10466605 TI - Nitric oxide synthase inhibition prevents intestinal damage in gastroschisis: a morphological evaluation in chick embryos. AB - PURPOSE: Increased small bowel nitric oxide (NO) synthase activity accused for postnatal intestinal dysmotility in gastroschisis. The purpose of this study is to evaluate the effect of prenatal NO synthase inhibition on intestinal damage in gastroschisis. METHODS: Sixteen-day-old fertilized chick eggs were divided into 4 groups. In the control group, the allantoic and amniotic membranes were opened to create a common cavity. In the gastroschisis group, a defect in the abdominal wall was made, and intestinal loops were exteriorized. In the gastroschisis pretreated with L-NAME group, gastroschisis was created, and L-NAME was administered into the amnioallantoic cavity for 4 days. In the gastroschisis sham pretreated group, after the same surgical procedure as the previous group, same amount of saline was given beside L-NAME. At the end of 20th day of incubation, intestinal morphological changes were investigated macroscopically and microscopically. RESULTS: Macroscopic changes such as shortening, thickening, and fibrous adhesions were found in the exteriorized bowels of the just gastroschisis group and the gastroschisis pretreated saline group. However, there was only mild thickening in the gastroschisis pretreated with L-NAME group. Microscopically, compared with the gastroschisis group, serosal thickness, muscular thickness, and bowel wall thickness were found to be significantly lower in the gastroschisis pretreated with L-NAME group (128.0 +/- 19.3 microm and 239.5 +/- 3.0 microm v 57.0 +/- 8.2 microm and 145.0 +/- 9.7 microm). CONCLUSION: It is possible to decrease intrauterine intestinal morphological changes in gastroschisis by inhibiting NO synthase. PMID- 10466606 TI - Outcome analysis for gastroschisis. AB - BACKGROUND/PURPOSE: Several factors are reportedly associated with an adverse outcome in gastroschisis, including mode of delivery, in utero diagnosis, type of closure, concurrent anomalies, intestinal atresia, and necrotizing enterocolitis (NEC). Since 1969, the authors have treated 185 patients who had gastroschisis. The authors analyzed their database to identify variables associated with increased morbidity and mortality. METHODS: A retrospective study of all patients with gastroschisis treated at our institution in the last 30 years was performed. The characteristics of survivors and nonsurvivors were compared. A logistic regression analysis was performed, with survival as the dependent variable, and the following parameters as independent variables: in utero diagnosis, mode of delivery, gestational age and birth weight, era of repair, type of closure, presence of other associated anomalies, intestinal atresia, and development of necrotizing enterocolitis. Further logistic regression analysis was performed, with various indicators of morbidity as dependent variables. These included development of sepsis, bowel obstruction, and complications related to the closure or to the silo. No attempt at long-term follow-up was made. RESULTS: A total of 185 infants with gastroschisis were treated at our institution from 1969 to 1999. Mean gestational age was 36.6 weeks, and the mean birth weight was 2,501 g. A total of 21 infants had intestinal atresia. NEC developed in 8 infants. Six infants had other serious anomalies. The overall survival rate was 91%. Survival improved in last 2 decades (94%). There were no differences in gestational age, birth weight and mode of delivery, method of closure, or presence of intestinal atresia between survivors and nonsurvivors. Only the era of repair (P = .002), presence of necrotizing enterocolitis (P = .044), and presence of other major anomalies (P < .001) correlated with mortality in the logistic regression analysis. Sepsis, bowel obstruction, and closure complications accounted for most of the morbidity. Analysis of these three morbidity factors identified low gestational age (P = .038) and development of necrotizing enterocolitis (P = .020) as independent predictors of sepsis. Closure complications were only associated with lower birth weight (P = .006). No predictors of bowel obstruction were identified. CONCLUSIONS: Mode of delivery, method of closure, birth weight and gestational age, and the presence of intestinal atresia do not appear to correlate with survival in infants with gastroschisis. Only the presence of another major anomaly, the era of repair, and the development of necrotizing enterocolitis were associated with increased mortality. Degree of prematurity and development of enterocolitis were associated with an increased incidence of septic complications. Low birth weight was a marker for closure complications. Type of delivery (vaginal or cesarean section) had no influence on either morbidity or mortality. PMID- 10466607 TI - Altered endothelin-3 and endothelin-B receptor mRNA expression in Hirschsprung's disease. AB - BACKGROUND: Recently, the endothelin-3 (EDN3) and endothelin-B receptor (EDNRB) gene have been recognized as susceptibility genes for Hirschsprung's disease (HD). However, gene mutations have been observed only in limited cases, and the role of EDN3 in the pathogenesis and motility dysfunction in HD is not understood fully. To evaluate the possible implication of EDN3 and EDNRB for the development of HD, we examined the EDN3 and EDNRB mRNA level in bowel specimens of HD patients. METHODS: Entire resected specimens of colon were obtained from 14 patients with HD. Eight age-matched control patients without gastroenteric disorders also were examined. mRNA was extracted from ganglionic and aganglionic segments of the HD specimens and normal colons. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate the relative amount of EDN3 and EDNRB mRNA. RESULTS: In normal colon, constant EDN3 and EDNRB mRNA expression was observed. In HD, EDN3 and EDNRB mRNA expression was observed. In HD, EDN3 and EDNRB mRNA levels were decreased both in ganglionic and aganglionic segment in 2 cases. In 6 cases, EDN3 mRNA expression was decreased in aganglionic segment and in another 2 cases, EDNRB mRNA expression was decreased in aganglionic segment. In the remaining 4 cases, EDN3 and EDNRB mRNA levels were similar to controls. CONCLUSION: The authors' findings indicate that loss of EDN3 and EDNRB function may be involved in the maldevelopment of neural crest-derived cells causing HD in many patients. PMID- 10466608 TI - Orthotopic liver transplantation for unresectable hepatoblastoma: a single center's experience. AB - BACKGROUND/PURPOSE: Complete surgical resection after chemotherapy is the definitive treatment for hepatoblastoma. However, orthotopic liver transplantation (OLT) is now accepted as a treatment modality for patients with unresectable tumours. The aim of this study was to review a single center's experience of OLT for unresectable hepatoblastoma. METHODS: A retrospective review of 8 patients with unresectable hepatoblastoma who were referred for liver transplantation was conducted. RESULTS: The patients assessed had an age range of 5 to 105 months at presentation; median, 24 months, (5 boys; 3 girls). Two patients have familial adenomatous polyposis, and one has right hemihypertrophy. All 8 patients had received standard chemotherapy according to SIOP (International Society of Pediatric Oncology) protocols. Extrahepatic metastases were found in 3 patients at diagnosis, but none had detectable metastases at the time of OLT. Four patients continued chemotherapy while awaiting OLT. Three patients received whole grafts, and 5 received reduced grafts. The median follow up period was 22 months (range, 2 to 78 months). Five patients are alive and well, although 1 patient had a second OLT for biliary cirrhosis secondary to biliary stricture at 6 years. Three patients died: one 26 days post OLT of sepsis and two of disease recurrence at 22 months and 70 months posttransplant. The actuarial survival rate is 88% and 65% at 1 and 5 years, respectively, whereas the overall survival rate is 62.5%. CONCLUSION: OLT for unresectable hepatoblastoma without extra hepatic metastases is highly successful with a low recurrence rate. PMID- 10466609 TI - Directing portal flow is essential for graft survival in auxiliary partial heterotopic liver transplantation in the dog. AB - BACKGROUND/PURPOSE: Auxiliary liver transplantation is an attractive alternative for orthotopic liver transplantation in patients with certain inborn errors of metabolism of the liver in which complete resection of the liver is unnecessary or even contraindicated. Because in these diseases portal hypertension is mostly absent, finding a balance in portal blood distribution between native liver and graft is complicated. The objective of this study was to investigate requirements for long-term (180 days) graft survival in auxiliary partial heterotopic liver transplantation (APHLT) in a dog model. METHODS: A metabolic defect was corrected in 26 dalmation dogs with a 60% beagle heterotopic auxiliary liver graft. Four groups of different portal inflow were studied. In the ligation group the portal vein to the host liver was ligated. In the split-flow group graft and host liver received separate portal inflow. In the banding group the distribution of the portal flow was regulated with an adjustable strapband and in the free-flow group the portal blood was allowed to flow randomly to host or graft liver. RESULTS: Metabolic correction increased in all groups after transplantation from 0.19 +/- 0.02 to 0.70 +/- 0.05 (P< .0001) but remained significantly better in the ligation and split-flow groups (graft survival, 135 +/- 27 and 144 +/- 31 days). In the banding group metabolic correction decreased significantly after 70 days, and although the grafts kept some function for 155 +/- 14 days, in 4 of 6 dogs portal thrombosis was found. In the free-flow group, competition for the portal blood led to reduced correction within 12 days and total loss of function in 96 +/- 14 days. Graft function also was assessed with technetium (Tc) 99m dimethyl iminodiacetic acid uptake. A good linear association between HIDA uptake and metabolic correction was observed (r = 0.74; P < .0005). Grafts that contributed more than 15% to the total uptake of HIDA showed biochemical correction. This indicates a critical graft mass of about 15% to 20% of the hepatocyte volume to correct this metabolic defect. CONCLUSION: Auxiliary partial heterotopic liver transplantation can be a valuable alternative treatment for inborn errors of hepatic metabolism if the native liver and the graft receive separate portal blood inflow. PMID- 10466610 TI - Mesenchymal hamartoma of the liver--new insight into histogenesis. AB - BACKGROUND/PURPOSE: Mesenchymal hamartoma (MH) of the liver is thought to develop from the ductal plates of the prenatal liver. This immunohistochemical study was performed to gain insight into the pathophysiology of its development. METHODS: Specimens from four MHs with adjacent liver, in one case from a biopsy and from the resected lesion after 6 years follow-up, were investigated with immunostaining on cryostatsectionswith antibodies against cytokeratins, vimentin, desmin and alpha-actin, as well as von Willebrand factor (factor VIII), fibroblast growth factor (FGF) receptors, FGF-1 (acidic FGF), FGF-2 (basic FGF), and the proliferation-associated Ki67 antigen. RESULTS: Fibrous tissue of MH stained positive not only for vimentin, but also for desmin and alpha-actin, whereas cytokeratins and factor VIII showed specific staining in biliary cysts and endothelial cells, respectively. All mesenchymal cells expressed proteins of the FGF receptor family. Although FGF-1 was only scarcely detectable, there was an accumulation of FGF-2 in borderline areas of liver to MH. Multiple Ki67 positive mesenchymal cells could be identified in these regions in all three MHs. However, we could not detect any proliferative activity in the MHs after follow up. CONCLUSIONS: The proliferative process in MH is still active during early childhood. FGF-2 may have a role in promoting this process. The positivity for desmin and alpha-actin of the lesions suggests that fat-storing (Ito) cells of the immature liver may be involved in the development of MH. PMID- 10466611 TI - The evaluation of technetium 99m-citrate scintigraphy in children with suspected appendicitis. AB - PURPOSE: The authors evaluated the use of technetium (Tc) 99m-citrate scan in 30 children whose diagnoses of appendicitis were unclear. METHODS: There were 17 boys and 13 girls (mean age 10.6 years). Nineteen of 30 patients had appendicitis confirmed at laparotomy and through histological examination, and 11 patients had other causes of acute abdominal pain. RESULTS: Children included in this study were similar with respect to age, duration of symptoms, temperature, white blood cell count, and the incidence of right lower quadrant tenderness. Sixteen children had positive scan for acute appendicitis, but 1 of them had false positive. Fourteen children had negative scan, but 4 of them had false-negative. The calculated values were 78.9% (15 of 19), 90.9% (10 of 11), 83.3% (25 of 30), 93.7% (15 of 16), and 71.4% (10 of 14) for sensitivity, specificity, accuracy, positive prediction, and negative prediction, respectively, in this study. CONCLUSION: The authors suggest the use of 99mTc-citrate scan in children when the diagnosis of appendicitis using other methods is unclear. PMID- 10466612 TI - Long-term follow-up of nineteen cystic lymphangiomas treated with fibrin sealant. AB - BACKGROUND: Surgical exeresis is regarded as the first choice treatment for cystic lymphangioma. Surgery may be extremely complex, giving rise to complications. Several therapeutic methods have been described to avoid the complications derived from the conventional surgical approach. The idea of treating lymphangioma by means of suction and injection of fibrin sealant (Tissucol), is an alternative to surgery. METHODS: The authors present 19 cases of cystic lymphangioma, treated with fibrin sealant injected into the lesion, during the 1991 to 1997 period. Two of the patients had been treated surgically and experienced recurrence of the tumor previous to treatment. In the other 17 cases, puncture was the only therapy applied. One patient required 3 punctures, another 6 patients required 2 punctures, and only 1 of them, after 2 unsuccessful punctures, was treated with surgical resection. The rest of the 10 cases subsided after the first puncture. Follow-up ranged between 3 and 72 months, with a mean of 40 months. RESULTS: The ultrasonographic (US) follow-up showed a complete remission in 17 patients treated with puncture. One patient remained with a small intermittent tumor, the appearance of which is related to catarrhal processes, and another patient rejected further puncture after the second one. No complications appeared. CONCLUSION: These results support the fact that the puncture, aspiration, followed by injection of Tissucol, is a choice in the surgical treatment of cystic lymphangioma. PMID- 10466613 TI - Fetal midgut volvulus presenting at term. AB - This report describes a rare case of intrauterine midgut volvulus that presented at term. The pregnancy was uncomplicated. There were no signs of fetal distress or polyhydramnios, and the child was delivered vaginally. This patient had the unusual presentation in that at the time of delivery, the patient was distended and acidotic. She immediately required an extensive resection of gangrenous small bowel. Comparing this case to the 10 other cases of fetal intestinal volvulus that have been reported, it seems this child was particularly fortunate that the volvulus occurred at a point in gestation when she was mature enough to tolerate birth and surgery. This case is also the first demonstration that volvulus can present with abdominal distension in the immediate newborn period. PMID- 10466614 TI - Repair of pediatric pectus deformity and congenital heart disease as a combined procedure. AB - Coexisting pectus deformity and congenital heart disease is not uncommon. Traditionally, the approach to this problem has been to repair each one with a separate surgical procedure because of fear of increased complications from bleeding, infections, and anesthesia. More recently, many reports of successful combined repair have been published, particularly in adults with coronary artery or aortic pathology. The authors wished to determine the feasibility of this combined procedure in younger patients, particularly those with a severe pectus deformity. Three patients underwent repair, including a 17 year old with Marfan's syndrome and a severe pectus excavatum deformity. The postoperative course was smooth for these patients, and all had good short- and long-term (over 18 months) results. PMID- 10466615 TI - A rare case of completely isolated duplication cyst of the alimentary tract. AB - A rare case of a gastrointestinal cystic duplication in a 7-day-old infant is described. The duplication diagnosed antenataly at 25 weeks of gestation was found during surgery to be separated from the gastrointestinal tract, hanging on a vascular pedicle, with no connection to the mesentery. The duplication was excised, and postoperative follow-up of 14 months was uneventful. The possible pathogenesis of this malformation is discussed. PMID- 10466616 TI - Gastrojejunal fistula after insertion of percutaneous endoscopic gastrostomy. AB - The authors report the case of a 12-year-old boy with cystic fibrosis, in whom a percutaneous endoscopic gastrostomy device migrated into the jejunum, forming a gastrojejunal fistula. PMID- 10466617 TI - An unusual foreign body in the bowel lumen causing obstruction in a neonate. AB - Most of the foreign bodies swallowed by children pass the entire gastrointestinal tract without any complication. Neonatal intestinal foreign bodies are extremely rare. A newborn with a small bowel obstruction caused by a fresh grape is reported. PMID- 10466618 TI - Use of a Palmaz stent for tracheomalacia: case report of an infant with esophageal atresia. AB - A male infant with congenital cardiac anomalies and esophageal atresia with tracheoesophageal fistula (EA-TEF) showed intractable respiratory symptoms after delayed primary repair of EA-TEF. Computed tomography demonstrated that the trachea was compressed by the enlarged aorta. Artificial ventilation was necessary even after aortopexy performed at 2 months of age. At 140 days of age, an expandable metallic stent (Palmaz stent) was inserted through a rigid bronchoscope into the trachea underfluoroscopic control. His respiratory status improved dramatically, and he was extubated in 18 hours. Although the follow-up period has been 9 months, the short-term result is satisfactory. The expandable metallic stent placement should be considered in patients with EA-TEF who show intractable respiratory symptoms caused by tracheomalacia. PMID- 10466619 TI - Ossifying fibromyxoid tumor of soft parts in a child: a case report. AB - Ossifying fibromyxoid tumor (OFMT) is a relatively rare tumor, most of which occur in adults with preferential localization in subcutis or muscles of the extremeties. Although Schwannian or cartilage origin has been suggested, histogenesis of this tumor still is unclear. The authors present a 6-year-old girl with retroperitoneal OFMT showing paraspinal extension, who is alive and tumor free 9 years after excision. The current case is the youngest reported patient showing unusual deep trunk site with surgically identified association with the spinal nerve. Both the clinical and histopathologic features strongly suggested Schwannian origin of this tumor. PMID- 10466620 TI - Duodenal duplication cyst manifested by duodeno-jejunal intussusception and hyperbilirubinemia. AB - A rare case of duodenal duplication cyst containing stones in a 17-year-old patient is presented. The cyst, acting as a leading point for duodeno-jejunal intussusception caused proximal small bowel obstruction and hyperbilirubinemia. Preoperative diagnosis was based on abdominal computerized tomography. At operation, the cyst wall was unroofed creating free drainage into the duodenal lumen without damaging the biliary and pancreatic ducts with resolution of symptoms. PMID- 10466621 TI - Bronchopexy for congenital bronchomalacia in the newborn. AB - Congenital bronchomalacia is a very unusual cause of respiratory distress in the newborn. The surgical management of this anomaly is challenging. The authors report on a newborn with congenital bronchomalacia successfully treated with bronchopexy. PMID- 10466622 TI - Necrotizing enterocolitis in omphalopagus conjoined twins. AB - Female omphalopagus twins underwent laparotomy on the second day of life after an antenatally diagnosed high jejunal bowel obstruction. Bowel resection and choldocho-enterostomies were performed. Despite recovery from laparotomy, the development of severe necrotizing enterocolitis (NEC) in one twin led to rapid deterioration and the death of both infants on day 34. The elucidation of the combined biliary tree, the dilemma of NEC in conjoined twins, and the possibilities of emergency separation are discussed. Consideration should be given to emergency separation of conjoined twins in the event of potentially lethal complications. PMID- 10466623 TI - Preoperative diagnosis of extralobar pulmonary sequestration with unusual vasculature: a case report. AB - Pulmonary sequestrations are uncommon congenital malformations of the lung. Vascular supply to pulmonary sequestrations is variable, with many unusual combinations. Adequate preoperative evaluation is hence advisable to avoid intraoperative complications. Magnetic resonance imaging (MRI) allows accurate diagnosis of extralobar pulmonary sequestration (ELPS), including definition of vascular supply. This may obviate the need for more invasive investigations that were suggested in the past. Presented here is a case of ELPS with its vascular attachments to the intercostal vessels. PMID- 10466624 TI - Systemic and local interferon gamma gene delivery to the lungs for treatment of allergen-induced airway hyperresponsiveness in mice. AB - Allergen-induced airway hyperresponsiveness, an animal model of asthma in humans, may respond to immunotherapy with Th1 cytokines. For example, local administration of recombinant IL-12 or IFN-gamma, or intratracheal delivery of the genes for these cytokines, has been shown to reduce the severity of allergen induced airway hyperresponsiveness (AHR) in rodent models. We reasoned that systemic cytokine gene delivery to the lungs by intravenous injection of lipid DNA complexes might also be an effective approach to treatment of allergen induced AHR. Therefore, the effects of either systemic or local pulmonary IFN gamma gene delivery were evaluated in mice with allergen-induced AHR. The effects of treatment on AHR, airway eosinophilia and cytokine production, and serum IgE concentrations were evaluated in mice that were first sensitized to ovalbumin and then subjected to aerosol ovalbumin challenge. Intravenous IFN-gamma gene delivery significantly inhibited development of AHR and airway eosinophilia and decreased serum IgE levels, compared with control mice or mice treated with noncoding DNA. Intratracheal IFN-gamma gene delivery also significantly inhibited AHR and airway eosinophilia, but did not affect serum IgE levels. Treatment with recombinant IFN-gamma was much less effective than IFN-gamma gene delivery by either route. We conclude that either systemic or local pulmonary delivery of a Th1 cytokine gene such as IFN-gamma may be an effective approach for treatment of allergen-induced asthma. PMID- 10466625 TI - Prevention of experimental allergic encephalomyelitis by intramuscular gene transfer with cytokine-encoding plasmid vectors. AB - Antiinflammatory cytokines such as transforming growth factor beta1 (TGF-beta1) and interleukin 4 (IL-4) can protect from autoimmune diseases. To study the immunoregulatory effects of these cytokines in vivo, we used a method of gene therapy that permits continuous cytokine delivery over a period of weeks. We injected naked plasmid DNA expression vectors encoding either TGF-beta1 (pVR-TGF beta1) or an IL-4-IgG1 chimeric protein (pVR-IL-4-IgG1) intramuscularly. This resulted in production of TGF-beta1 or IL-4-IgG1, respectively, and protection from myelin basic protein (MBP)-induced experimental allergic encephalomyelitis (EAE). TGF-beta1 gene delivery had pronounced downregulatory effects on T cell proliferation and production of interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha), on in vitro restimulation with MBP. IL-4-IgG1 vector administration also suppressed these responses, although much less than TGF beta1, and enhanced secretion of endogenous IL-4. Therapy resulted in a significant decrease in the severity of histopathologic inflammatory lesions. In the CNS, treatment with either vector suppressed IL-12 and IFN-gamma mRNA expression, while IL-4 and TGF-beta1 mRNA levels were increased compared with control mice. Thus, cytokine plasmid treatment appeared to inhibit MBP-specific pathogenic Thl responses, while enhancing endogenous secretion of protective cytokines. We demonstrate that gene therapy with these vectors is an effective therapeutic strategy for EAE. PMID- 10466626 TI - Restoration of bacterial killing activity of human respiratory cystic fibrosis cells through cationic vector-mediated cystic fibrosis transmembrane conductance regulator gene transfer. AB - In vitro and in vivo studies have demonstrated that gene transfer of the CFTR (cystic fibrosis transmembrane conductance regulator) cDNA into human respiratory cells through nonviral vectors can occur safely and can be done repeatedly. Although functional evaluation of CFTR in cystic fibrosis (CF) patients enrolled in phase I clinical trials using cationic liposomes has shown a partial correction of nasal potential difference, a biological assay indicating a therapeutic relevance of CFTR gene transfer is still missing. Our aims were to study the induction of killing activity toward Pseudomonas aeruginosa (PA) in CF cells by cationic vector-mediated CFTR gene transfer and to use this assay as a therapeutic end point. Luciferase expression and GFP FACS analysis were used to evaluate the optimal vector and the efficiency of gene transfer into non-CF human respiratory cells growing from nasal polyp explants at the air-liquid interface. To prove that transgenic CFTR was expressed in CF cell cultures under the same experimental conditions, a specific RT-PCR was performed. Challenge of the outgrowths with a known amount of PA showed a bacterial clearance activity by non CF respiratory cells, while in the case of CF cells it even resulted in bacterial growth. Cationic vector-mediated CFTR cDNA determined the recovery of bacterial clearance activity only under those conditions yielding 5% or more of GFP positive cells. The results shown in this study might be helpful in considering cationic vectors as therapeutic nonviral vectors for transferring CFTR into human CF respiratory cells, as well as for restoring the bacterial killing activity defective in cystic fibrosis. PMID- 10466627 TI - Circulating alpha-galactosidase A derived from transduced bone marrow cells: relevance for corrective gene transfer for Fabry disease. AB - Fabry disease is caused by a deficiency of the lysosomal enzyme alpha galactosidase A (alpha-gal A). We previously engineered a retrovirus encoding human alpha-gal A and demonstrated enzymatic correction of patient cells. Further, we demonstrated metabolic cooperativity, in that corrected cells secrete alpha-gal A that can be taken up and utilized by bystander cells in vitro. In the present study, we created a system to examine and quantitate this phenomenon in vivo. To differentiate from endogenous alpha-gal A, we constructed a retroviral vector (pUMFG/alpha-gal A/FLAG) containing a fusion form of alpha-gal A with a specific tag sequence at the carboxy terminus. The catalytic activity of the fusion protein was identical to wild-type alpha-gal A. The fusion protein was overexpressed in and secreted by transduced patient cells. In uptake studies, the fusion protein was detected in the lysosome-enriched fraction of recipient cells. We then examined the effectiveness of the pUMFG/alpha-g A/FLAG retroviral vector in vivo. Murine bone marrow (BM) cells were transduced and transplanted into irradiated hosts. After 9 weeks, proviral DNA was detected by PCR in peripheral blood and BM mononuclear cells. More importantly, specific fusion protein enzymatic activity could be demonstrated in those cells and in plasma. Thus, we have demonstrated that overexpressed alpha-gal A enters the circulation from transduced BM cells and is stable over a significant period of time. PMID- 10466628 TI - Expression of chimeric granulocyte-macrophage colony-stimulating factor/interleukin 2 receptors in human cytotoxic T lymphocyte clones results in granulocyte-macrophage colony-stimulating factor-dependent growth. AB - Adoptive immunotherapy with ex vivo-expanded antigen-specific cytotoxic T lymphocytes (CTLs) has been shown to clear viral infections and eliminate tumors in murine models. Clinical trials have also reported promising data for the use of adoptive immunotherapy to treat cytomegalovirus (CMV) and Epstein-Barr viral (EBV) infections in bone marrow transplant recipients. For these indications, the need for ex vivo-expanded CTLs is often short lived, until the immune system is reconstituted by the donor transplant. In chronic disease settings, increased longevity of adoptively transferred CTLs and generation of memory will be necessary. The additional administration of helper functions normally supplied by antigen-specific T helper (Th) cells will probably be essential for long-term survival of adoptively transferred CTLs. Toward this goal of supplying helper functions, we transduced human CTLs with chimeric GM-CSFR/IL-2R receptors that deliver an IL-2 signal on binding GM-CSF. Clones expressing the chimeric receptors proliferated in response to GM-CSF. Stimulation with antigen induced GM CSF production and resulted in an autocrine growth loop such that the CTL clones proliferated in the absence of exogenous cytokines. This type of genetic modification has potential for increasing the circulating half-life and, by extension, the efficacy of ex vivo-expanded CTLs. PMID- 10466629 TI - Engraftment of gene-marked hematopoietic progenitors in myeloma patients after transplant of autologous long-term marrow cultures. AB - We conducted a phase I hematopoietic stem cell (HSC) gene-marking trial in patients undergoing autologous blood or marrow stem cell transplant for the treatment of multiple myeloma. Between 500 and 1000 ml of bone marrow was harvested from each of 14 myeloma patients and 1 syngeneic donor. A mean of 3.3x10(9) cells per patient were plated in 20 to 50 long-term marrow culture (LTMC) flasks and maintained for 3 weeks. LTMCs were exposed on days 8 and 15 to clinical-grade neo(r)-containing retrovirus supernatant (G1Na). A mean of 8.23x10(8) day-21 LTMC cells containing 5.2x10(4) gene-marked granulocyte macrophage progenitor cells (CFU-GM) were infused along with an unmanipulated peripheral blood stem cell graft into each patient after myeloablative therapy. Proviral DNA was detected in 71% of 68 tested blood and bone marrow samples and 150 of 2936 (5.1%) CFU-GM derived from patient bone marrow samples after transplant. The proportion of proviral DNA-positive CFU-GM declined from a mean of 9.8% at 3 months to a mean of 2.3% at 24 months postinfusion. Southern blots of 26 marrow and blood samples were negative. Semiquantitative PCR analysis indicated that gene transfer was achieved in 0.01-1% of total bone marrow and blood mononuclear cells (MNCs). Proviral DNA was also observed in EBV-transformed B lymphocytes, in CD34+ -enriched bone marrow cells, and in CFUs derived from the latter progenitors. Gene-modified cells were detected by PCR in peripheral blood and bone marrow for 24 months after infusion of LTMC cells. Sensitivity and specificity of the PCR assays were independently validated in four laboratories. Our data confirm that HSCs may be successfully transduced in stromal based culture systems. The major obstacle to therapeutic application of this approach remains the overall low level of genetically modified cells among the total hematopoietic cell pool in vivo. PMID- 10466630 TI - Improved titers of retroviral vectors from the human FLYRD18 packaging cell line in serum- and protein-free medium. AB - The influence of serum on the production of retroviral vectors by the HT1080 human fibrosarcoma-derived packaging cell line FLYRD18 was investigated. A fourfold increase in virus titer was observed under serum-free conditions, as compared with medium supplemented with 10% fetal calf serum. A similar improvement was also seen for bulk transduction efficiency. Serum had a negative and dose-dependent effect on titer without affecting cell growth, virus stability, or infectivity. In contrast to virus from NIH 3T3-derived packaging cells [Hanenberg, H., et al. (1996). Nature Med. 2, 876-882], the FLYRD18-derived virus did not adhere to fibronectin or serum proteins adsorbed at the surface of culture flasks. Electron microscopy supports the conclusion that the effect of serum is at the level of virus production by the cells. Addition of soybean trypsin inhibitor had an inhibitory effect on virus production, while pretreatment of serum with trypsin was found to enhance the retroviral titer. These results suggest that protease inhibitors present in serum may be responsible for the inhibition of virus production. The exact mechanism remains, however, to be determined. As compared with medium supplemented with 10% serum, the combination of increased virus titer and absence of exogenous protein under serum-free conditions resulted in a 300-fold increase in the virus:total protein ratio in the supernatants harvested from the FLYRD18 packaging line. This improvement enhances prospects for further concentration and purification of the virus. PMID- 10466631 TI - Basic study on gene therapy of human malignant glioma by use of the cationic multilamellar liposome-entrapped human interferon beta gene. AB - For gene therapy of human malignant glioma, we adopted positively charged multilamellar liposomes entrapping the human interferon beta (hIFN-beta) gene. One week after the transplantation of human malignant glioma U251-SP cells to produce glioma in nude mouse brain, the liposomes entrapping the gene (500 ng of DNA per 25 nmol of lipids per 2 microl) were injected into the same site of the cell transplantation once every second day for a total of five injections; and by this means the tumor completely disappeared. To confirm the antiproliferative effect of hIFN-beta, we performed an in vitro study using a plasmid containing a secretion signal sequence-deleted hIFN-beta gene and one containing the hIFN-beta gene inserted in reverse. In both cases, there was no hIFN-beta release into the medium and no growth inhibition effect. On addition of anti-hIFN-beta antibody to the medium, the growth inhibition effect was abolished. As this cell line expresses IFN-alpha/beta receptor, the hIFN-beta produced in the transfected cells could be released and acted in a paracrine manner. For 120 days the body weight change of normal mice treated by the same procedure as used in the curing experiment was not significant among the groups injected with empty liposomes, plasmid only, and liposomes entrapping the gene. In all of these three groups, death, abnormal behavior, and significant histological changes were not observed. PMID- 10466632 TI - A universal granulocyte-macrophage colony-stimulating factor-producing bystander cell line for use in the formulation of autologous tumor cell-based vaccines. AB - Irradiated tumor cells transduced with the gene encoding the cytokine GM-CSF have been extensively studied as a vaccine formulation capable of priming systemic antitumor immune responses in the tumor-bearing host. In spite of the therapeutic promise of this vaccine strategy demonstrated in both animal models and early phase clinical trials, clinical development has been limited by difficulties pertaining to the need to establish in culture the tumor of each patient and to perform individualized gene transfer. To circumvent these issues, we generated an HLA-negative human cell line producing large quantities of human GM-CSF for use as a universal bystander cell to be mixed with unmodified autologous tumor cells in the formulation of a vaccine. This line is easily propagated as a suspension culture in defined, serum-free medium. In a mouse model, we find that vaccination with a mixture of autologous tumor cells and an MHC-negative allogeneic GM-CSF producing bystander cell primes antitumor immune responses that are equivalent or better than those achieved using autologous tumor cells directly transduced to secrete GM-CSF. This strategy greatly simplifies further clinical development of autologous tumor cell-based vaccines. PMID- 10466633 TI - Enzyme/prodrug therapy for head and neck cancer using a catalytically superior cytosine deaminase. AB - The use of cytosine deaminase (CD) in conjunction with 5-fluorocytosine (5-FC) has been studied for cancer gene therapy as a means of achieving tumor-specific generation of the toxic metabolite 5-fluorouracil (5-FU). Since 5-FC is frequently used as an antifungal agent, and because it has little or no efficacy as an antibacterial agent, we hypothesized that yeast CD (YCD) might be more efficient at utilizing 5-FC as a substrate and hence be a better choice for a CD/5-FC gene therapy strategy than the typically utilized bacterial CD (BCD). To that end Saccharomyces cerevisiae CD was cloned from yeast genomic DNA and expressed in vitro. Functional analysis of BCD and YCD expressed in COS-1 cells indicated that BCD and YCD both utilized cytosine with equal efficacy; however, 5 FC was an extremely poor substrate for BCD, with an apparent catalytic efficiency 280-fold lower than that observed for YCD. Retroviral infection of tumor cell lines in vitro indicated that the IC50 of 5-FC was 30-fold lower in YCD-infected cultures as compared with cultures infected with BCD retrovirus. In addition, when SCCVII murine squamous cell carcinoma cells were infected in vitro at low rates of infection (< or =10%) there was no significant cytotoxicity toward BCD expressing cells while there was potent cytotoxicity to both YCD-expressing cells and "bystander cells" even at this low level of expression. Finally, stable BCD- or YCD-expressing SCCVII clones were developed and used in an orthotopic immune competent model of head and neck cancer. Subsequent treatment with 5-FC followed by monitoring of tumor growth by noninvasive magnetic resonance imaging (MRI) and survival of animals indicated a growth delay during the course of 5-FC treatment for BCD-expressing tumors, which quickly regrew at the end of treatment. In contrast, YCD-expressing tumors exhibited not only a growth delay, which was of longer duration, but also in some cases frank tumor regression and complete cures occurred. PMID- 10466634 TI - Improved titers for helper virus-free herpes simplex virus type 1 plasmid vectors by optimization of the packaging protocol and addition of noninfectious herpes simplex virus-related particles (previral DNA replication enveloped particles) to the packaging procedure. AB - A helper virus-free herpes simplex virus type 1 (HSV-1) plasmid vector system has potential for both gene therapy and physiological studies, but relatively low titers have complicated use of this system. In this article, the packaging efficiency was improved by optimizing the packaging protocol and by adding noninfectious HSV-1-related particles, i.e., previral DNA replication enveloped particles (PREPs), during the packaging procedure. PREPs contain many of the tegument proteins that are thought to enhance an HSV-1 infection. Use of both the optimized packaging protocol and the PREPs resulted in an approximately 50-fold increase in the titer, and five different HSV-1 vectors were packaged using this procedure. A purified vector stock (7.8x10(8) infectious vector particles/ml) was microinjected into the striatum, the rats were sacrificed 4 days after gene transfer, and the brains were found to contain an average of approximately 6740 X Gal-positive striatal cells. This improved packaging procedure may augment use of this vector system. PMID- 10466635 TI - A simple method for constructing E1- and E1/E4-deleted recombinant adenoviral vectors. AB - We previously developed a two-plasmid in vitro ligation method that did not require a recombination step to produce new recombinant E1- or E1/E3-deleted adenoviral vectors. In this study, we have modified the system to improve the simplicity of vector construction and, in addition, to allow for production of an E1/E4-deleted vector. PMID- 10466636 TI - A phase I/II study of hepatic artery infusion with wtp53-CMV-Ad in metastatic malignant liver tumours. AB - Colorectal cancer (CRC) is the second commonest cause of cancer death in the UK, with greater than 40% of these patients destined to die of the disease despite current medical management. Death is commonly due to liver metastases with sequelae including progressive liver dysfunction. Most patients with liver metastases present with tumours that are unresectable and incurable with existing therapies. The median survival for CRC patients after diagnosis with liver metastases is approximately 6 months or less. The human p53 gene is a tumour suppressor gene involved in the control of cell proliferation. Loss of wild-type p53 function is associated with the uncontrolled growth of many types of human cancers. The reintroduction and expression of wild-type p53 into p53 altered tumour cells has been shown to suppress tumour growth or induce apoptosis in both in vitro and in vivo models. In our experience greater than 50% of CRC tumours have p53 alterations. This study seeks to evaluate the safety, biological efficacy and the effectiveness of wtp53-CMV-Ad treatment which is a recombinant adenoviral vector containing the wild-type human p53 gene. It will be administered by infusion via the hepatic artery, for the regional gene therapy of malignant liver tumours. Study patients will have incurable metastatic (CRC) malignant tumours of the liver with evidence of p53 alteration in their liver tumours. In vitro studies have demonstrated p53-specific antiproliferative effects of wtp53-CMV-Ad on human liver tumour cells and in vivo studies have demonstrated p53-specific antiproliferative effects on human liver tumour cells. The vector Ad-p53 is a recombinant, replication-defective adenovirus based on adenovirus serotype 5. It contains a sequence encoding wild-type p53 whose expression is under the control of the human cytomegalovirus immediate early promoter-enhancer. This construct will be growth in 293 cells which contain the adenoviral E1A and E1B coding sequences which have been removed from the vector to render it replication defective. The study design is an open-label, non randomised, single-dose, dose escalation Phase I/II clinical trial anticipated to involve a maximum of 19 patients. wtp53-CMV-Ad will be administered by infusion in a reservoir connected to the hepatic artery, for regional gene therapy (surgically implanted pump) in 3 escalating doses to successive cohorts of 3 patients each until the maximum tolerated dose is determined. Subsequently, 10 patients will be treated with this dose. Regional wtp53-CMV-Ad therapy will be administered as a single bolus infusion via hepatic artery catheter. The route of administration of wtp53-CMV-Ad via hepatic artery infusion is designed to maximise gene therapy exposure to the malignant tumours while minimising exposure to normal tissues outside the liver. The clinical protocol is designed to monitor treatment toxicity. Another objective is to evaluate the biological efficacy, including efficiency and stability of gene transfer by analysis of tumour tissues following therapy. As an important part of this objective the pharmacokinetics of wtp53-CMV-Ad will be studied. Clinical evidence of anti-tumour efficacy will also be collected. In addition, the safety and efficacy of different doses levels of wtp53-CMV-Ad will be studied. PMID- 10466637 TI - Human gene marker/therapy clinical protocols. PMID- 10466638 TI - Five rings of Musashi. PMID- 10466639 TI - The use of heparin in patients in whom a pulmonary embolism is suspected after total hip arthroplasty. AB - BACKGROUND: The morbidity and mortality associated with pulmonary embolism are well known, as is the benefit of the use of heparin in patients who have a pulmonary embolism. However, the patterns of heparin use as well as its undesirable effects, especially in patients who have recently had a total hip arthroplasty, have been less well studied. Thus, concern arises regarding the use of heparin in patients who have no firm evidence of a pulmonary embolism. The purpose of the current study was to track the use of heparin and associated orthopaedic complications in patients in whom a pulmonary embolism was suspected after a total hip arthroplasty. METHODS: The records of 150 patients in whom a pulmonary embolism had been suspected after a total hip arthroplasty were reviewed retrospectively. The rates of individual complications (such as stroke, infection, and hematoma) and those of groups of complications (such as medical complications, orthopaedic complications, and all complications combined) were recorded and then were stratified according to the treatment (with or without heparin), the presence or absence of pulmonary embolism, and other variables. RESULTS: Thirty-two (47 percent) of sixty-eight patients who were managed with heparin had complications compared with sixteen (20 percent) of eighty-two patients who were not thus managed (p = 0.0006). Specifically, patients who were managed with heparin were more likely to have gastrointestinal bleeding, hematological complications, a loose prosthesis, a hematoma, or an early revision arthroplasty (p<0.05 for all). With the numbers available, the use of heparin was not found to be significantly associated with an increased risk of death, stroke, or infection at the site of the prosthesis. Interestingly, thirty-one (31 percent) of ninety-nine patients who had ventilation-perfusion scans that demonstrated normal findings or findings indicating a low probability of pulmonary embolism were given heparin before the diagnosis of a pulmonary embolism was excluded, and sixteen (52 percent) of these thirty-one had complications. CONCLUSIONS: Given this risk profile, we advise against the use of heparin before the diagnosis of pulmonary embolism is established in patients who have had a total hip arthroplasty. This recommendation is supported by algorithms, in widely read medical texts, pertaining to the use of heparin in patients in whom a pulmonary embolism is suspected. PMID- 10466640 TI - Nonunion after periprosthetic femoral fracture associated with total hip arthroplasty. AB - BACKGROUND: Nonunion after a periprosthetic femoral fracture associated with total hip arthroplasty occurs rarely. There is little information, to our knowledge, regarding the prevalence of this complication, its treatment, and the functional outcomes of treatment. The purpose of this study was to identify the patterns and frequency of nonunions of femoral fractures around total hip prostheses and to evaluate the results and problems associated with treatment of this complication in a consecutive series of patients. METHODS: The study included twenty-three nonunions of periprosthetic femoral fractures in twenty three patients with an average age of fifty-five years (range, twenty-two to eighty-five years) at the time of the initiation of treatment of the nonunion. Thirteen of the fractures occurred during or after a primary total hip arthroplasty, and ten occurred during or after a revision total hip arthroplasty. According to the classification system of Duncan and Masri, there were six B1 fractures (associated with a well fixed prosthesis), seven B2 fractures (associated with a loose stem), and ten B3 fractures (associated with very poor proximal bone). Ten patients were managed with revision to a long-stem prosthesis. Six patients had revision to a proximal femoral replacement prosthesis. A two-stage technique consisting of removal of the prosthesis and open reduction and internal fixation of the nonunion followed by reimplantation of the prosthesis was used in two patients. Two patients were managed initially with bone-grafting alone, and two patients were managed nonoperatively. One patient who had an infection at the site of the nonunion was managed definitively with resection arthroplasty. RESULTS: The duration of clinical follow-up averaged 8.3 years (range, three months to twenty-three years), and that of radiographic surveillance averaged 7.0 years (range, eight months to seventeen years). Of the thirteen patients in whom an attempt to achieve union was made and for whom radiographs were available, nine eventually had bone-healing. Five of the twenty three femora became infected and were treated with resection arthroplasty. Of the seventeen patients who had not had a resection arthroplasty for infection and for whom radiographs were available at the time of the most recent follow-up, eleven had a stable and well fixed implant and six had a loose implant as seen radiographically or had had a revision because of aseptic loosening. Seventeen patients had no or mild pain at the time of the most recent follow-up, but ten required two-handed support to walk. The overall complication rate was 52 percent (twelve of twenty-three patients). CONCLUSIONS: Nonunion of a femoral fracture associated with a total hip prosthesis is an infrequent problem. Treatment is difficult, with a high rate of complications and relatively poor functional outcomes. The data from this series must be interpreted with caution, as patients were managed over a period of three decades and many did not have the advantage of modern techniques of revision hip arthroplasty. Prevention of nonunion by optimum treatment of the initial fracture is most important. Treatment of a femoral nonunion about a total hip implant should be implemented on the basis of the status of the fixation of the prosthesis and the quality of the surrounding bone. PMID- 10466641 TI - Impaction allografting with cement for revision of the femoral component. A minimum four-year follow-up study with use of a precoated femoral stem. AB - BACKGROUND: Cancellous impaction allografting with cement for revision of the femoral component has conventionally been performed with a polished, tapered implant, which was designed to allow subsidence of the component. However, subsidence has been associated with pain in the thigh, dislocation of the hip, and revision of the component. This prospective study tested the hypothesis that good clinical results can be achieved--without subsidence of the component--with use of impaction allografting and a precoated, collared, straight stem for difficult femoral revisions. METHODS: Twenty-nine patients had revision of the femoral component with use of impaction allografting with cement and a Harris Precoat stem. Impaction allografting was performed when loss of metaphyseal and diaphyseal bone precluded revision with more straightforward techniques or when reconstitution of bone was considered a specific goal of the reconstruction (as was sometimes the case with revision of the component in younger patients). The patients were followed prospectively and were evaluated with use of the Harris hip score and serial radiographs. The patients were followed for a minimum of four years (mean, sixty-three months), except for four who died. RESULTS: Four patients died before the minimum four-year follow-up period had elapsed; all four had the prosthesis in place at the time of death. The Harris hip scores improved from a preoperative mean of 54 points (poor) (range, 21 to 91 points) to a mean of 87 points (good) (range, 41 to 100 points) at the time of the most recent follow-up. Kaplan-Meier survivorship analysis, with aseptic loosening as the end point, was 92 percent (95 percent confidence interval, 82 to 100 percent) at six years; one additional hip failed because of a hematogenous infection at seventy three months postoperatively, for an overall failure rate of 12 percent (three of twenty-five patients) at the time of the most recent follow-up. Two hips needed a repeat revision; one was revised because of subsidence of the stem with recurrent osteolysis and the other, in a patient who had hemodialysis, because of late sepsis. A third femoral component subsided and failed but was not revised. Radiographic evidence of bone-stock reconstitution was observed in six (29 percent) of the twenty-one patients for whom radiographs were available. As in other series of patients managed with impaction allografting, the complication rate was high; excluding the revisions, three reoperations were performed, and six patients had either intraoperative femoral fracture or perforation necessitating cerclage wiring or cortical strut allografting and cerclage wiring at the time of the procedure. There were six nonunions in eighteen patients who had been operated on with a transtrochanteric approach. CONCLUSIONS: Difficult revisions of the femoral component with use of impaction allografting and a precoated stem provided satisfactory clinical and radiographic results at the time of intermediate-term follow-up. However, the high rate of complications in our series led us to refine our indications for the procedure. PMID- 10466642 TI - Intra-articular fractures of the distal aspect of the radius: arthroscopically assisted reduction compared with open reduction and internal fixation. AB - BACKGROUND: There is no consensus that an arthroscopically guided operation can improve the anatomical and functional results of treatment of intra-articular fractures of the distal aspect of the radius. The purpose of the present prospective study was to determine the usefulness of arthroscopically assisted reduction of displaced intra-articular fractures of the distal aspect of the radius by comparing the results of that procedure with those of conventional open reduction and internal fixation. METHODS: Thirty-four fractures were treated with arthroscopically guided reduction with use of one volar and two dorsal arthroscopic portals. The fractures were pinned, and external fixation was used with or without autogenous bone graft. Intraoperative fluoroscopy was not used. Forty-eight fractures were treated with conventional open reduction and internal fixation with a plate and screws or with pinning, with or without external fixation. The average duration of follow-up for all fractures was thirty-one months. RESULTS: The scores for overall outcome, assessed with use of the system of Gartland and Werley and that of Green and O'Brien as modified by Cooney et al., demonstrated that the group that had had an arthroscopically assisted procedure had better outcomes than the group that had had conventional open reduction and internal fixation. The group that had had an arthroscopically assisted procedure also had significantly better ranges of flexion-extension and radial-ulnar deviation of the wrist and grip strength (p<0.05). We detected an association between the maximum step and gap displacement and evidence of osteoarthritis of the radiocarpal joint (p<0.001), but we did not find a significant association, with the numbers available, between the scores for osteoarthritis, graded according to the scale of Knirk and Jupiter, and the scores for overall outcome, assessed with the scale of Gartland and Werley and the modified system of Green and O'Brien, in either group (p = 0.376). The radiographic results showed that the patients who had had an arthroscopically assisted procedure had better reduction of volar tilt, ulnar variance, and articular (gap) displacement than did those who had been managed with conventional open reduction and internal fixation (p<0.05 for each comparison). CONCLUSIONS: An arthroscopically guided operation achieved an accurate reduction of intra-articular fractures of the distal aspect of the radius. Minimum capsular and adjacent soft-tissue scarring reduced postoperative contracture, which improved the overall functional results. We recommend arthroscopically guided reduction and internal fixation not only for young adults but for all patients who are less than seventy years old and have an intra-articular fracture of the distal part of the radius with more than one millimeter of displacement on plain radiographs. PMID- 10466643 TI - Osteoid osteoma of the elbow: a diagnostic challenge. AB - BACKGROUND: Osteoid osteoma is a painful benign neoplasm that is rarely found in the elbow region. METHODS: The study included fourteen patients, and we believe that this is the largest reported series of patients with osteoid osteoma of the elbow evaluated at one institution. Most of the patients had had symptoms for a prolonged period and had had multiple invasive procedures before an accurate diagnosis was made. Although findings on physical examination generally are nonspecific and are not always accurate in localizing the lesion, plain tomograms and computed tomography scans were most helpful in identifying the nidus in the present study. Thirteen of the patients had limited motion of the elbow before the definitive diagnosis was made, and ten of these thirteen had a mean flexion contracture of 38 degrees. RESULTS: Removal of the nidus resulted in relief of pain and improvement in the range of motion of the elbow in all fourteen patients. A persistent postoperative flexion contracture was more common in the patients who had had a previous arthrotomy of the elbow than in those who had not had that procedure. CONCLUSIONS: It is important to recognize this uncommon entity to avoid the morbidity associated with a prolonged delay in diagnosis. Because the symptoms resolve after excision of the lesion, the surgeon can avoid unnecessary soft-tissue dissection and release of the contracture. PMID- 10466644 TI - Transfer of the latissimus dorsi muscle after failed repair of a massive tear of the rotator cuff. A two to five-year review. AB - INTRODUCTION: Seventeen patients with an average age of fifty-five years (range, thirty-two to seventy-seven years) who had ongoing pain and impaired function following failed operative treatment of a massive tear of the rotator cuff were managed with a transfer of the latissimus dorsi muscle as a salvage operation. METHODS: The patients were examined at an average of fifty-one months (range, twenty-four to seventy-two months) after the operation. Pain, function, and satisfaction were assessed with use of a questionnaire, visual analog and ordinal scales, physical examination, and the University of California at Los Angeles shoulder score. RESULTS: Fourteen of the seventeen patients were found to have significant relief of pain (p<0.0001) and a significant improvement in function (p<0.001 for all activities except lifting more than fifteen pounds [6.8 kilograms], for which the p value was <0.0036) and were satisfied with the result of the operative procedure. Fifteen patients stated that they would have the operative procedure again under similar circumstances. Seven of eight patients with a detached or nonfunctional anterior portion of the deltoid had substantial improvement. Three operations were classified as failures because the patients were not satisfied with the result and had ongoing pain and impaired function. All three failures were in patients who had a work-related injury. Overall, six patients had a work-related injury, and only three of them had a satisfactory result. There were three complications, all related to contracture of a hypertrophic axillary scar. CONCLUSIONS: The results in this series indicate that transfer of the latissimus dorsi muscle is a reasonable approach for salvage after failed operative treatment of a massive tear of the rotator cuff. PMID- 10466645 TI - Ipsilateral total shoulder and elbow arthroplasties in patients who have rheumatoid arthritis. AB - BACKGROUND: The data on seventeen patients with rheumatoid arthritis who had been managed with ipsilateral total shoulder and elbow arthroplasties were analyzed to determine whether the operative technique, the presence of total shoulder and total elbow prostheses in the same upper extremity, or complications of the arthroplasties affected the result in each joint or the overall functional outcome of the upper extremity. METHODS: Seventeen patients with rheumatoid arthritis who were managed with a total of eighteen ipsilateral total shoulder and elbow arthroplasties were evaluated. The most recent physical examination was at an average of six years and six months (range, two years and one month to fourteen years) postoperatively. Radiographs, including 40-degree oblique and axillary radiographs of the shoulder as well as anteroposterior and lateral radiographs of the elbow, were made at an average of six years and eleven months (range, two years and two months to twenty-two years and eleven months) postoperatively. The radiographs of the shoulder were examined for loosening of the glenoid component, glenohumeral subluxation, and radiolucency at the bone cement or bone-implant interface. The functional results of the total shoulder arthroplasties were evaluated with use of the rating systems of Neer et al. and Cofield. The Mayo elbow-performance score was used to evaluate elbow function. A rating system was also developed to assess the overall function of the upper extremity, including pain and motion of both the elbow and the shoulder. With this system, the overall function of the upper extremity was rated as excellent, good, fair, or poor. RESULTS: Evaluation of the shoulders revealed substantial relief of pain and an increase in active elevation. On radiographic evaluation, eight glenoid and five humeral components were considered to be loose. There were no reoperations. According to the rating system of Neer et al., eight shoulders had a satisfactory result and eight had an unsatisfactory result with limited active abduction. Limited-goals rehabilitation was successful after one shoulder arthroplasty and unsuccessful after another. There were two type-B periprosthetic humeral fractures. There was also substantial relief of pain in the elbows as well as an increase in the extension-flexion arc; the pronation-supination arc was sufficient for tasks of daily living. There was no radiographic loosening. Two elbows had an avulsion of the triceps, and two had aseptic loosening (one of which also had a worn bushing); all four needed a reoperation. One other elbow had persistent ulnar neuritis. The average interval between the arthroplasties was two years and eight months when the shoulder was replaced first and three years and five months when the elbow was replaced first. The interval between the joint replacements and the sequence of the joint replacements were not found to influence the outcome. Function of the extremity was improved by replacement of either the shoulder or the elbow alone; however, it improved significantly only when both joints were replaced (p = 0.03). According to combined clinical outcomes scores, there were nine excellent outcomes, four good outcomes, four fair outcomes, and one poor outcome after ipsilateral total shoulder and elbow arthroplasties. CONCLUSIONS: When there is severe arthritis of both the shoulder and the elbow, consideration should be given to replacing both joints in order to obtain optimum functional and clinical outcomes. The possibility of fracture of the humeral shaft necessitates an alteration of the technique for ipsilateral total shoulder and elbow arthroplasties. PMID- 10466646 TI - Complications and functional outcomes of reconstruction with an osteoarticular allograft after intra-articular resection of the proximal aspect of the humerus. AB - BACKGROUND: The purpose of this study was to evaluate the functional outcome and the complications of reconstruction with an osteoarticular allograft in patients who had had intra-articular resection of the proximal aspect of the humerus. METHODS: Sixteen patients who had had intra-articular resection and reconstruction of the proximal aspect of the humerus for the treatment of a tumor between 1986 and 1996 were evaluated. The length of the resected part of the humerus ranged from eight to 27.5 centimeters. The resections were classified as either S34A or S345A resections of the shoulder girdle on the basis of the Musculoskeletal Tumor Society classification system. Reconstruction was performed with use of a nonirradiated, frozen osteoarticular allograft with intact capsular and rotator cuff attachments. Dual orthogonal dynamic compression plates were used for internal fixation of the allograft to the host bone. The oncological parameters that were evaluated included survival of the patient, local recurrence, and metastasis. The radiographic parameters included time to union, stability of the joint, fracture of the allograft, and fragmentation of the epiphysis of the allograft (subchondral collapse). Survival of the graft was assessed with Kaplan-Meier survival analysis. The modified Musculoskeletal Tumor Society evaluation system was used to assess functional outcome. RESULTS: At a median of forty-seven months (range, fourteen to 130 months) after the operation, fourteen of the patients in the study group were free of disease and two had died of disease. No patient had local recurrence or nonunion. Late complications included four fractures of the allograft and one infection of the graft. A Kaplan Meier survival curve demonstrated a 68 percent rate of survival of the allograft at five years. Instability of the glenohumeral joint in the form of ptosis and anterior subluxation was noted in three patients, and dislocation of the glenohumeral joint was seen in eight patients. On the basis of the modified Musculoskeletal Tumor Society functional evaluation, the mean score at the most recent follow-up evaluation (at a mean of thirty-four months) was 70 percent. This score was lower than the mean score of 81 percent at a mean of fourteen months. All patients had normal manual dexterity and had mild or no pain at the most recent follow-up evaluation. However, all had restriction of recreational activities or partial disability in addition to limitations with regard to placement of the hand and the ability to lift. CONCLUSIONS: Reconstruction of the proximal aspect of the humerus with an osteoarticular allograft is an option that provides good relief of pain and preserves manual dexterity. However, in our study, function was limited by impairment of elevation of the shoulder and hand as well as by decreased strength of the shoulder. There was an extremely high rate of complications, including joint instability, fracture of the allograft, and infection of the allograft. We no longer routinely perform this reconstruction at our institution. PMID- 10466647 TI - Subluxation of the talocalcaneal joint in adults who have symptomatic flatfoot. AB - BACKGROUND: When flatfoot is acquired during adulthood, the shape of the foot changes. In addition to a decreased arch, there may be valgus angulation of the hindfoot or abduction of the forefoot, or both. However, there is little objective information to provide a better understanding of the anatomical or morphological changes that occur in acquired adult flatfoot. We wondered if such an understanding of the three-dimensional anatomy might shed light on the pathway by which these changes occur. We designed this study to measure the three dimensional position of the talocalcaneal joint in patients who have painful flatfoot. METHODS: Computed tomography scans of the feet of eight patients who had symptomatic flatfoot were used to construct a model of the talocalcaneal articulation. The scans were performed on a custom loading frame developed to simulate weight-bearing with the foot in a neutral position while a seventy-five newton axial compressive load was applied. The digital data from the scans were used to make three-dimensional computer models of the articular surfaces of the talus and calcaneus of each foot. These models then were used to calculate the percentage of the articular surface that was in contact and, conversely, the percentage that was subluxated. Two surfaces were modeled for each bone; the posterior facet formed one surface, and the anterior and middle facets were combined to form the second surface. The data were compared, with use of Mann Whitney nonparametric U analysis, with those derived from scans of the feet of four patients without a deformity of the hindfoot who served as controls. RESULTS: A mean (and standard deviation) of 68+/-9 percent of the posterior facet of the calcaneus was in contact with the talus in the patients who had flatfoot compared with 92+/-2 percent in the controls, and a mean of 51+/-23 percent of the anterior and middle facets of the calcaneus was in contact with the talus in the patients who had flatfoot compared with 95+/-6 percent in the controls. These differences were significant (p = 0.0066 for both). CONCLUSIONS: Marked subluxation of the talocalcaneal joint occurs in some patients who have symptomatic planoabductovalgus deformity. PMID- 10466648 TI - Pathophysiological effect of fat embolism in a canine model of pulmonary contusion. AB - BACKGROUND: The objective of this study was to determine the individual and combined effects of pulmonary contusion and fat embolism on the hemodynamics and pulmonary pathophysiology in a canine model of acute traumatic pulmonary injury. METHODS: After a thoracotomy, twenty-one skeletally mature dogs were randomly assigned to one of three groups. Unilateral pulmonary contusion alone was produced in Group 1 (seven dogs); pulmonary contusion and fat embolism, in Group 2 (seven dogs); and fat embolism alone, in Group 3 (seven dogs). Pulmonary contusion was produced by standardized compression of the left lung with a piezoelectric force transducer. Fat embolism was produced by femoral and tibial reaming followed by pressurization of the intramedullary canals. Cardiac output, systolic blood pressure, peak airway pressure, pulmonary arterial pressure, pulmonary capillary wedge pressure, partial pressure of arterial oxygen, and partial pressure of carbon dioxide were monitored for all groups. From these data, several outcome parameters were calculated: total thoracic compliance, alveolar-arterial oxygen gradient, and ratio of partial pressure of arterial oxygen to fractional inspired oxygen concentration. All of the dogs were killed after eight hours, and tissue samples were obtained from the brain, kidneys, and lungs for histological analysis. Lung samples were assigned scores for pulmonary edema (the presence of fluid in the alveoli) and inflammation (the presence of neutrophils or hyaline membranes, or both). The percentage of the total area occupied by fat was determined. RESULTS: Pulmonary contusion alone caused a significant increase in the alveolar-arterial oxygen gradient but only after seven hours (p = 0.034). Fat embolism alone caused a significant transient decrease in systolic blood pressure (p = 0.001) and a significant transient increase in pulmonary arterial pressure (p = 0.01) and pulmonary capillary wedge pressure (p = 0.015). Fat embolism alone also caused a significant sustained decrease in the ratio of partial pressure of arterial oxygen to fractional inspired oxygen concentration (p = 0.0001) and a significant increase in the alveolar-arterial oxygen gradient (p = 0.0001). The combination of pulmonary contusion and fat embolism caused a significant transient increase in pulmonary capillary wedge pressure (p = 0.0013) as well as a significant sustained decrease in partial pressure of arterial oxygen (p = 0.0001) and a significant decrease in systolic blood pressure (p = 0.001) that lasted for an hour. Pulmonary contusion followed by fat embolism caused a significant increase in peak airway pressure (p = 0.015), alveolar-arterial oxygen gradient (p = 0.0001), and pulmonary arterial pressure (p = 0.01), and these effects persisted for five hours. Total thoracic compliance was decreased 6.4 percent by pulmonary contusion alone, 4.6 percent by fat embolism alone, and 23.5 percent by pulmonary contusion followed by fat embolism. The ratio of partial pressure of arterial oxygen to fractional inspired oxygen concentration was decreased 23.7 percent by pulmonary contusion alone, 52.3 percent by fat embolism alone, and 65.8 percent by pulmonary contusion followed by fat embolism. The mean pulmonary edema score was significantly higher with the combined injury than with either injury alone (p = 0.0001). None of the samples from the lungs demonstrated inflammation. Fat embolism combined with pulmonary contusion resulted in a significantly greater mean percentage of the area occupied by fat in the noncontused right lung than in the contused left lung (p = 0.001); however, no significant difference between the right and left lungs could be detected with fat embolism alone. The mean percentage of the glomerular and cerebral areas occupied by fat was greater with fat embolism combined with pulmonary contusion than with fat embolism alone (p = 0.0001 and p = 0.01, respectively). (ABSTRACT TRUNCATED) PMID- 10466649 TI - Osteonecrosis of the lateral aspect of the talar dome after triple arthrodesis. A report of three cases. PMID- 10466650 TI - Hemolytic disease of the newborn caused by transfusion of a husband's directed blood donation. A case report. PMID- 10466651 TI - Acquired flatfoot in adults due to dysfunction of the posterior tibial tendon. PMID- 10466652 TI - Closed reduction and tendon transfer for treatment of dislocation of the glenohumeral joint secondary to brachial plexus birth palsy. PMID- 10466653 TI - The treatment of symptomatic os acromiale. PMID- 10466654 TI - Thought for safety of aid workers in dangerous places. PMID- 10466655 TI - New diagnostic criteria for diabetes--are they doing what they should? PMID- 10466656 TI - Risk of neurodegenerative diseases in children conceived by intracytoplasmic sperm injection? PMID- 10466657 TI - Immersion deaths and swim failure--implications for resuscitation and prevention. PMID- 10466658 TI - Brachial palsies from obstetric procedures. PMID- 10466659 TI - Meningococcal conjugate vaccines: new opportunities and new challenges. PMID- 10466660 TI - Bright future for nuclear medicine. PMID- 10466661 TI - Glucose tolerance and mortality: comparison of WHO and American Diabetes Association diagnostic criteria. The DECODE study group. European Diabetes Epidemiology Group. Diabetes Epidemiology: Collaborative analysis Of Diagnostic criteria in Europe. AB - BACKGROUND: The American Diabetes Association (ADA) recommend that fasting glucose alone with the oral glucose tolerance test should be used to diagnose diabetes mellitus. We assessed mortality associated with the ADA fasting-glucose criteria compared with the WHO 2 h post-challenge glucose criteria. METHODS: We assessed baseline data on glucose concentrations at fasting and 2 h after the 75 g oral glucose tolerance test from 13 prospective European cohort studies, which included 18,048 men and 7316 women aged 30 years or older. Mean follow-up was 7.3 years. We assessed the risk of death according to the different diagnostic glucose categories. FINDINGS: Compared with men who had normal fasting glucose (< 6.1 mmol/L), men with newly diagnosed diabetes mellitus by the ADA fasting criteria (> or = 7.0 mmol/L) had a hazard ratio for death of 1.81 (95% CI 1.49 2.20); for women the hazard ratio was 1.79 (1.18-2.69). For impaired fasting glucose (6.1-6.9 mmol/L), the hazard ratios were 1.21 (1.05-1.41) and 1.08 (0.70 1.66). For the WHO criteria (> or = 11.1 mmol/L), the ratios for newly diagnosed diabetes were 2.02 (1.66-2.46) in men and 2.77 (1.96-3.92) in women, and for impaired glucose tolerance (7.8-11.1 mmol/L) were 1.51 (1.32-1.72) and 1.60 (1.22 2.10). Within each fasting-glucose classification, mortality increased with increasing 2 h glucose. However, for 2 h glucose classifications of impaired glucose tolerance, and diabetes, there was no trend for increasing fasting glucose concentrations. INTERPRETATION: Fasting-glucose concentrations alone do not identify individuals at increased risk of death associated with hyperglycaemia. The oral glucose tolerance test provides additional prognostic information and enables detection of individuals with impaired glucose tolerance, who have the greatest attributable risk of death. PMID- 10466662 TI - Cardiovascular disease in older adults with glucose disorders: comparison of American Diabetes Association criteria for diabetes mellitus with WHO criteria. AB - BACKGROUND: The new fasting American Diabetes Association (ADA) criteria for the diagnosis of diabetes mellitus rely mainly on fasting blood glucose concentrations and use a lower cut-off value for diagnosis than the WHO criteria. We aimed to assess the sensitivity of these criteria for the detection of cardiovascular disease, the main complication of diabetes mellitus in the elderly. METHODS: We did a cross-sectional and prospective analysis of 4515 participants of the Cardiovascular Health Study, an 8 year longitudinal study designed to identify factors related to the onset and course of cardiovascular disease in adults aged at least 65 years. We calculated the prevalence and incidence of cardiovascular disease for the ADA and WHO criteria. FINDINGS: There was a higher prevalence of cardiovascular disease among individuals with impaired glucose or newly diagnosed diabetes by both criteria than among those with normal glucose concentrations. However, because fewer individuals had abnormal glucose states by the fasting ADA criteria (22.3%) than by the WHO criteria (46.8%), the number of cases of cardiovascular disease attributable to abnormal glucose states was a third of that attributable by the WHO criteria (53 vs 159 cases per 10,000). For the two sets of criteria, the relative risk for incident cardiovascular disease (mean follow-up 5.9 years) was higher in individuals with impaired glucose and newly diagnosed diabetes than in those with normal glucose. Individuals classified as normal by the fasting ADA criteria had a higher absolute number of incident events (455 of 581 events) than those classified as normal by the WHO criteria (269 of 581 events). Fasting ADA criteria were therefore less sensitive than the WHO criteria for predicting cardiovascular disease among individuals with abnormal glucose (sensitivity, 28% vs 54%). INTERPRETATION: The new fasting ADA criteria seem to be less predictive than the WHO criteria for the burden of cardiovascular disease associated with abnormal glucose in the elderly. PMID- 10466663 TI - Immersion deaths and deterioration in swimming performance in cold water. AB - BACKGROUND: General hypothermia (deep body temperature <35 degrees C) has been implicated in immersion-related deaths, but many deaths occur too quickly for it to be involved. We investigated changes in swimming capability in cold water to find out whether such changes could lead to swim failure and drowning. METHODS: Ten volunteers undertook three self-paced breaststroke swims in a variable-speed swimming flume, in water at 25 degrees C, 18 degrees C, and 10 degrees C, for a maximum of 90 min. During each swim, we measured oxygen consumption, rectal temperature, swim speed and angle, and stroke rate and length. Swim failure was defined as being unable to keep feet off the bottom of the flume. FINDINGS: All ten swimmers completed 90 min swims at 25 degrees C, eight completed swims at 18 degrees C, and five at 10 degrees C. In 10 degrees C water, one swimmer reached swim failure after 61 min and four were withdrawn before 90 min with rectal temperatures of 35 degrees C when they were close to swim failure. Swimming efficiency and length of stroke decreased more and rate of stroke and swim angle increased more in 10 degrees C water than in warmer water. These variables seemed to characterise impending swim failure. INTERPRETATION: Impaired performance and initial cardiorespiratory responses to immersion probably represent the major dangers to immersion victims. Consequently, treatment should be aimed at symptoms resulting from near-drowning rather than severe hypothermia. PMID- 10466664 TI - Azithromycin in control of trachoma. AB - BACKGROUND: Trachoma is the leading cause of preventable blindness. Programmes to prevent blindness due to trachoma are based on community-wide treatment with topical tetracycline. We assessed the potential of community-wide azithromycin treatment for trachoma control. METHODS: Pairs of villages in trachoma endemic areas of Egypt, The Gambia, and Tanzania were matched on trachoma rates in 1-10 year-old children. Villages were randomly assigned community-wide oral azithromycin treatment (three doses with intervals of 1 week) or treatment with 1% topical tetracycline (once daily for 6 weeks). Clinical examinations were done at baseline, 2-4.5 months, and 12-14 months after treatment. Chlamydia trachomatitis was identified by ligase chain reaction (LCR). Analyses were by intention to treat. Univariate comparisons and multivariate analyses were used to compare outcomes. FINDINGS: LCR positivity was correlated with clinical severity, but about 30% of Egyptian and Gambian villagers with no active disease were LCR positive. Village-wide LCR positivity ranged from 16.5% (Tanzania) to 43.6% (Egypt). Treatment compliance was over 90% except in the tetracycline treatment village in Egypt. Of the participants initially LCR positive, 866 (95%) of 924 who received at least one azithromycin dose and 482 (82%) of 587 who received 28 days or more topical tetracycline, were negative at follow-up. At 1 year, village wide LCR positivity rates were substantially lower than at baseline with both treatments; the decreases were greater with azithromycin than with tetracycline (93% vs 77% in Egypt, 78 vs 66% in The Gambia, 64 vs 55% in Tanzania). Similarly, greater reduction in clinical activity occurred after azithromycin. In multivariate analyses, factors associated with being LCR positive at 1 year were: not receiving azithromycin; age under 10 years; and LCR positivity at baseline. INTERPRETATION: Community-wide treatment with oral azithromycin markedly reduces C. trachomatis infection and clinical trachoma in endemic areas and may be an important approach to control of trachoma. PMID- 10466665 TI - Non-pathogenic Escherichia coli versus mesalazine for the treatment of ulcerative colitis: a randomised trial. AB - BACKGROUND: Ulcerative colitis has been suggested to be caused by infection and there is circumstantial evidence linking Escherichia coli with the condition. Our aim was to find out whether the administration of a non-pathogenic strain of E. coli (Nissle 1917) was as effective as mesalazine in preventing relapse of ulcerative colitis. We also examined whether the addition of E. coli to standard medical therapy increased the chance of remission of active ulcerative colitis. METHODS: This was a single-centre, randomised, double-dummy study in which 120 patients with active ulcerative colitis were invited to take part. 116 patients accepted; 59 were randomised to mesalazine and 57 to E. coli. All patients also received standard medical therapy together with a 1-week course of oral gentamicin. After remission, patients were maintained on either mesalazine or E. coli and followed up for a maximum of 12 months. A two-stage, conditional, intention-to-treat analysis was done. FINDINGS: 44 (75%) patients in the mesalazine group attained remission compared with 39 (68%) in the E. coli group. Mean time to remission was 44 days (median 42) in the mesalazine group and 42 days (median 37) for those treated with E. coli. In the mesalazine group, 32 (73%) patients relapsed compared with 26 (67%) in the E. coli group. Mean duration of remission was 206 days in the mesalazine group (median 175) and 221 days (median 185) in the E. coli group. INTERPRETATION: Our results suggest that treatment with a non-pathogenic E. coli has an equivalent effect to mesalazine in maintaining remission of ulcerative colitis. The beneficial effect of live E. coli may provide clues to the cause of ulcerative colitis. PMID- 10466666 TI - Linkage between male infertility and trinucleotide repeat expansion in the androgen-receptor gene. AB - BACKGROUND: Androgens acting via the androgen receptor bring about stimulation and maintenance of spermatogenesis. If mutations in the androgen-receptor gene interfere with the receptor's function, this effect may partly account for impaired spermatogenesis. We aimed to find out whether expansion of a trinucleotide repeat in the androgen-receptor gene is associated with male infertility. METHODS: We analysed 67 coded semen and blood samples from a predominantly white group of male infertility patients and controls. Clinical analyses included cause of infertility, sperm count, and reproductive hormone concentrations. Analysis of trinucleotide (CAG) repeat length and point mutations in the androgen-receptor gene was done by PCR, single-stranded conformational polymorphism, and DNA sequencing. FINDINGS: Screening and characterisation of the androgen-receptor gene in 35 patients and 32 controls showed no point mutations in the gene. 30 of the infertile patients had idiopathic azoospermia or oligozoospermia, and these men had significantly longer CAG repeat tracts than controls (mean 23.2 [SE 0.7] vs 20.5 [0.3], p=0.0001). The odds of having CAG repeat lengths of 20 were six-fold higher for fertile men than for men with a spermatogenic disorder. INTERPRETATION: Our results indicate a relation between CAG repeat length in the androgen-receptor gene and the risk of defective spermatogenesis. With the use of intracytoplasmic sperm injection, this mutation could be inherited, possibly leading to an increase in male infertility in future generations. Should further elongation of the CAG repeat occur in these future generations, there is an added risk of increased severity of male infertility, and potentially an increased incidence of neurodegenerative disease. PMID- 10466667 TI - One swollen hand. PMID- 10466668 TI - Fetal brain activity demonstrated by functional magnetic resonance imaging. AB - Functional magnetic resonance imaging was used to study fetal brain activity. This activity was in response to an auditory stimulus. PMID- 10466669 TI - In-vivo demonstration of dopaminergic degeneration in dementia with Lewy bodies. AB - With the dopaminergic presynaptic ligand FP-CIT and single photon emission tomography we have shown a severe dopaminergic degeneration in a patient with a necropsy confirmed diagnosis of dementia with Lewy bodies (DLB). We suggest that functional imaging of the nigrostriatal dopamine pathway helps to distinguish DLB from Alzheimer's disease during life. PMID- 10466670 TI - Why is coeliac disease endemic in the people of the Sahara? AB - The prevalence of antiendomysial antibody (AEA) in 989 Saharawi children was 5.6%. Intestinal biopsies in a subsample confirmed that AEA is a marker of coellac disease in people living in a developing country. PMID- 10466671 TI - Ovarian cancer and infertility: a genetic link? AB - A genetic mechanism may be responsible for the increased incidence of ovarian cancer in some infertile women (ie, those who failed to conceive despite treatment). PMID- 10466672 TI - Antibodies to lipopolysaccharides of Escherichia coli serogroups O5 and O165 in healthy adults. AB - The observation that over 50% of healthy blood donors have serum antibodies to the lipopolysaccharide of Escherichia coli O5 and O165 influences the serodiagnosis of infection with verocytotoxin-producing E. coli. PMID- 10466673 TI - Reduction of serum uric acid by hormone replacement therapy in postmenopausal women with hyperuricaemia. AB - Reduction of serum uric acid by hormone replacement therapy in postmenopausal women with hyperuricaemia is one of the cardiovascular protective mechanisms by which hormone replacement therapy reduces the risk of cardiovascular disease. PMID- 10466674 TI - Prophylactic antibiotics and endocervical microbial inoculation of the endometrium at embryo transfer. AB - Prophylactic antibiotics administered routinely to women at the time of oocyte retrieval were associated with a reduction in positive microbiology cultures of embryo catheter tips 48 h later in 78.4% of patients. The implantation and clinical pregnancy rates were significantly lower (9.3% vs 21.6%, p<0.001; 18.7% vs 41.3%, p<0.01) in the women with positive microbial catheter-tip cultures. PMID- 10466676 TI - Raloxifene reduces spinal fractures postmenopausally. PMID- 10466675 TI - Centenarians: the older you get, the healthier you have been. AB - A retrospective analysis of the health and functional status of centenarians shows they are healthy and independent for most of their lives and experience a relatively rapid terminal decline. PMID- 10466677 TI - Plaque detection: the key to tackling atherosclerosis? PMID- 10466678 TI - Drug rationing comes out into the open in the UK. PMID- 10466679 TI - Pickets force pregnancy-advice centre to close. PMID- 10466680 TI - Southern African AIDS agreement. PMID- 10466681 TI - Public protests after rape in Pakistani hospital. PMID- 10466682 TI - Indian government puts welfare of monkeys before biomedical research. PMID- 10466683 TI - Italy enforces EU directive on specialist practice. PMID- 10466684 TI - UK hospital pays dearly to cut waiting lists. PMID- 10466685 TI - Cardiological applications of nuclear medicine. AB - Cardiovascular mortality is falling in most industrialised nations. Primarily responsible for this encouraging trend are preventive measures such as risk factor modification but improved medical and surgical management have helped too. Clinical decision making in the patient with coronary heart disease demands techniques that not only describe coronary anatomy but also provide functional indices for early detection and to monitor the severity and extent of disease. Nuclear medicine methods can characterise non-invasively myocardial function, perfusion, and metabolism. Novel radiopharmaceuticals, improvements in imaging equipment, and extensive validation have contributed to the growing clinical acceptance of these techniques and to their cost-effective integration in the workup of patients with cardiovascular disease. PMID- 10466686 TI - Pemphigus and bullous pemphigoid. AB - Pemphigus and bullous pemphigoid are distinct autoimmune blistering diseases that are characterised by the presence of autoantibodies directed against specific adhesion molecules of the skin and mucous membranes. The comparison and contrast of molecular mechanism of blister formation of these two diseases provide a rational diagnostic and therapeutic approach to affected patients. PMID- 10466687 TI - Charles Dickens' work to help establish Great Ormond Street Hospital, London. PMID- 10466688 TI - "I wish I'd got my new ears last year, mum". PMID- 10466689 TI - Breast cancer in women with palpable breast cysts. PMID- 10466690 TI - Breast cancer in women with palpable breast cysts. PMID- 10466691 TI - Breast cancer in women with palpable breast cysts. PMID- 10466692 TI - Audio-computer interviewing to measure HIV-risk behaviour. PMID- 10466693 TI - Making AIDS a business imperative. PMID- 10466694 TI - Postdural-puncture headache. PMID- 10466695 TI - Outbreak of Coca-Cola-related illness in Belgium: a true association. PMID- 10466696 TI - Outbreak of Coca-Cola-related illness in Belgium: a true association. PMID- 10466697 TI - Benefit of beta-blockers for heart failure. PMID- 10466698 TI - Wound closure. PMID- 10466699 TI - Causality assessment of adverse drug effects: when is rechallenge ethically acceptable? PMID- 10466700 TI - Debate continues on recombinant bovine somatotropin. PMID- 10466701 TI - Health risks of genetically modified foods. PMID- 10466702 TI - Scientific and research culture in Mexico. PMID- 10466703 TI - Increased medical indemnities in Ireland. PMID- 10466704 TI - Paying respect to organs. PMID- 10466705 TI - Estimation of missing values. PMID- 10466706 TI - Albumin and the Medicines Control Agency. PMID- 10466707 TI - Hard to swallow. PMID- 10466708 TI - The Nobel Chronicles. 1968: Har Khorana (b 1922); Robert Holley (1922-93); Marshall Nirenberg (b 1927). PMID- 10466710 TI - No time to hide. PMID- 10466709 TI - The difference between science and dogma. PMID- 10466711 TI - Annotation competition spurs Drosophila sequencing efforts. PMID- 10466712 TI - Kansas kicks evolution out the classroom. PMID- 10466714 TI - What price 'prestige' in publishing? PMID- 10466713 TI - Monsanto rapped for misleading press advertisements. PMID- 10466715 TI - Icelanders opt out of genetic database. PMID- 10466716 TI - Icelanders opt out of genetic database. PMID- 10466717 TI - Phytochromes. Tripping the light fantastic. PMID- 10466718 TI - Mast-cell heparin demystified. PMID- 10466719 TI - Cognition by a mini brain. PMID- 10466720 TI - Chemokine control of HIV-1 infection. PMID- 10466721 TI - A movement-sensitive area in auditory cortex. PMID- 10466722 TI - Context generalization in Drosophila visual learning requires the mushroom bodies. AB - The world is permanently changing. Laboratory experiments on learning and memory normally minimize this feature of reality, keeping all conditions except the conditioned and unconditioned stimuli as constant as possible. In the real world, however, animals need to extract from the universe of sensory signals the actual predictors of salient events by separating them from non-predictive stimuli (context). In principle, this can be achieved if only those sensory inputs that resemble the reinforcer in their temporal structure are taken as predictors. Here we study visual learning in the fly Drosophila melanogaster, using a flight simulator, and show that memory retrieval is, indeed, partially context independent. Moreover, we show that the mushroom bodies, which are required for olfactory but not visual or tactile learning, effectively support context generalization. In visual learning in Drosophila, it appears that a facilitating effect of context cues for memory retrieval is the default state, whereas making recall context-independent requires additional processing. PMID- 10466723 TI - Synaptic function modulated by changes in the ratio of synaptotagmin I and IV. AB - Communication within the nervous system is mediated by Ca2+-triggered fusion of synaptic vesicles with the presynaptic plasma membrane. Genetic and biochemical evidence indicates that synaptotagmin I may function as a Ca2+ sensor in neuronal exocytosis because it can bind Ca2+ and penetrate into lipid bilayers. Chronic depolarization or seizure activity results in the upregulation of a distinct and unusual isoform of the synaptotagmin family, synaptotagmin IV. We have identified a Drosophila homologue of synaptotagmin IV that is enriched on synaptic vesicles and contains an evolutionarily conserved substitution of aspartate to serine that abolishes its ability to bind membranes in response to Ca2+ influx. Synaptotagmin IV forms hetero-oligomers with synaptotagmin I, resulting in synaptotagmin clusters that cannot effectively penetrate lipid bilayers and are less efficient at coupling Ca2+ to secretion in vivo: upregulation of synaptotagmin IV, but not synaptotagmin I, decreases evoked neurotransmission. These findings indicate that modulating the expression of synaptotagmins with different Ca2+-binding affinities can lead to heteromultimers that can regulate the efficiency of excitation-secretion coupling in vivo and represent a new molecular mechanism for synaptic plasticity. PMID- 10466724 TI - Interaction of glutamic-acid-rich proteins with the cGMP signalling pathway in rod photoreceptors. AB - The assembly of signalling molecules into macromolecular complexes (transducisomes) provides specificity, sensitivity and speed in intracellular signalling pathways. Rod photoreceptors in the eye contain an unusual set of glutamic-acid-rich proteins (GARPs) of unknown function. GARPs exist as two soluble forms, GARP1 and GARP2, and as a large cytoplasmic domain (GARP' part) of the beta-subunit of the cyclic GMP-gated channel. Here we identify GARPs as multivalent proteins that interact with the key players of cGMP signalling, phosphodiesterase and guanylate cyclase, and with a retina-specific ATP-binding cassette transporter (ABCR), through four, short, repetitive sequences. In electron micrographs, GARPs are restricted to the rim region and incisures of discs in close proximity to the guanylate cyclase and ABCR, whereas the phosphodiesterase is randomly distributed. GARP2, the most abundant splice form, associates more strongly with light-activated than with inactive phosphodiesterase, and GARP2 potently inhibits phosphodiesterase activity. Thus, the GARPs organize a dynamic protein complex near the disc rim that may control cGMP turnover and possibly other light-dependent processes. Because there are no similar GARPs in cones, we propose that GARPs may prevent unnecessary cGMP turnover during daylight, when rods are held in saturation by the relatively high light levels. PMID- 10466725 TI - Increased affiliative response to vasopressin in mice expressing the V1a receptor from a monogamous vole. AB - Arginine vasopressin influences male reproductive and social behaviours in several vertebrate taxa through its actions at the V1a receptor in the brain. The neuroanatomical distribution of vasopressin V1a receptors varies greatly between species with different forms of social organization. Here we show that centrally administered arginine vasopressin increases affiliative behaviour in the highly social, monogamous prairie vole, but not in the relatively asocial, promiscuous montane vole. Molecular analyses indicate that gene duplication and/or changes in promoter structure of the prairie vole receptor gene may contribute to the species differences in vasopressin-receptor expression. We further show that mice that are transgenic for the prairie vole receptor gene have a neuroanatomical pattern of receptor binding that is similar to that of the prairie vole, and exhibit increased affiliative behaviour after injection with arginine vasopressin. These data indicate that the pattern of V1a-receptor gene expression in the brain may be functionally associated with species-typical social behaviours in male vertebrates. PMID- 10466726 TI - Heparin is essential for the storage of specific granule proteases in mast cells. AB - All mammals produce heparin, a negatively charged glycosaminoglycan that is a major constituent of the secretory granules of mast cells which are found in the peritoneal cavity and most connective tissues. Although heparin is one of the most studied molecules in the body, its physiological function has yet to be determined. Here we describe transgenic mice, generated by disrupting the N deacetylase/N-sulphotransferase-2 gene, that cannot express fully sulphated heparin. The mast cells in the skeletal muscle that normally contain heparin lacked metachromatic granules and failed to store appreciable amounts of mouse mast-cell protease (mMCP)-4, mMCP-5 and carboxypeptidase A (mMC-CPA), even though they contained substantial amounts of mMCP-7. We developed mast cells from the bone marrow of the transgenic mice. Although these cultured cells contained high levels of various protease transcripts and had substantial amounts of mMCP-6 protein in their granules, they also failed to express mMCP-5 and mMC-CPA. Our data show that heparin controls, through a post-translational mechanism, the levels of specific cassettes of positively charged proteases inside mast cells. PMID- 10466727 TI - Abnormal mast cells in mice deficient in a heparin-synthesizing enzyme. AB - Heparin is a sulphated polysaccharide, synthesized exclusively by connective tissue-type mast cells and stored in the secretory granules in complex with histamine and various mast-cell proteases. Although heparin has long been used as an antithrombotic drug, endogenous heparin is not present in the blood, so it cannot have a physiological role in regulating blood coagulation. The biosynthesis of heparin involves a series of enzymatic reactions, including sulphation at various positions. The initial modification step, catalysed by the enzyme glucosaminyl N-deacetylase/N-sulphotransferase-2, NDST-2, is essential for the subsequent reactions. Here we report that mice carrying a targeted disruption of the gene encoding NDST-2 are unable to synthesize sulphated heparin. These NDST-2-deficient mice are viable and fertile but have fewer connective-tissue type mast cells; these cells have an altered morphology and contain severely reduced amounts of histamine and mast-cell proteases. Our results indicate that one site of physiological action for heparin could be inside connective-tissue type mast cells, where its absence results in severe defects in the secretory granules. PMID- 10466728 TI - The chemokine receptor CCR4 in vascular recognition by cutaneous but not intestinal memory T cells. AB - Lymphocytes that are responsible for regional (tissue-specific) immunity home from the blood to the intestines, inflamed skin or other sites through a multistep process involving recognition of vascular endothelial cells and extravasation. Chemoattractant cytokine molecules known as chemokines regulate this lymphocyte traffic, in part by triggering arrest (stopping) of lymphocytes rolling on endothelium. Here we show that many systemic memory T cells in blood carry the chemokine receptor CCR4 and therefore respond to its ligands, the chemokines TARC and MDC. These cells include essentially all skin-homing cells expressing the cutaneous lymphocyte antigen and a subset of other systemic memory lymphocytes; however, intestinal (alpha4beta7+) memory and naive T cells respond poorly. Immunohistochemistry reveals anti-TARC reactivity of venules and infiltration of many CCR4+ lymphocytes in chronically inflamed skin, but not in the gastrointestinal lamina propria. Moreover, TARC induces integrin-dependent adhesion of skin (but not intestinal) memory T cells to the cell-adhesion molecule ICAM-1, and causes their rapid arrest under physiological flow. Our results suggest that CCR4 and TARC are important in the recognition of skin vasculature by circulating T cells and in directing lymphocytes that are involved in systemic as opposed to intestinal immunity to their target tissues. PMID- 10466729 TI - Binding of phytochrome B to its nuclear signalling partner PIF3 is reversibly induced by light. AB - The phytochrome photoreceptor family directs plant gene expression by switching between biologically inactive and active conformers in response to the sequential absorption of red and farred photons. Several intermediates that act late in the phytochrome signalling pathway have been identified, but fewer have been identified that act early in the pathway. We have cloned a nuclear basic helix loop-helix protein, PIF3, which can bind to non-photoactive carboxy-terminal fragments of phytochromes A and B and functions in phytochrome signalling in vivo. Here we show that full-length photoactive phytochrome B binds PIF3 in vitro only upon light-induced conversion to its active form, and that photoconversion back to its inactive form causes dissociation from PIF3. We conclude that photosensory signalling by phytochrome B involves light-induced, conformer specific recognition of the putative transcriptional regulator PIF3, providing a potential mechanism for direct photoregulation of gene expression. PMID- 10466730 TI - Nucleosome mobilization catalysed by the yeast SWI/SNF complex. AB - The generation of a local chromatin topology conducive to transcription is a key step in gene regulation. The yeast SWI/SNF complex is the founding member of a family of ATP-dependent remodelling activities capable of altering chromatin structure both in vitro and in vivo. Despite its importance, the pathway by which the SWI/SNF complex disrupts chromatin structure is unknown. Here we use a model system to demonstrate that the yeast SWI/SNF complex can reposition nucleosomes in an ATP-dependent reaction that favours attachment of the histone octamer to an acceptor site on the same molecule of DNA (in cis). We show that SWI/SNF-mediated displacement of the histone octamer is effectively blocked by a barrier introduced into the DNA, suggesting that this redistribution involves sliding or tracking of nucleosomes along DNA, and that it is achieved by a catalytic mechanism. We conclude that SWI/SNF catalyses the redistribution of nucleosomes along DNA in cis, which may represent a general mechanism by which ATP-dependent chromatin remodelling occurs. PMID- 10466731 TI - Four-helical-bundle structure of the cytoplasmic domain of a serine chemotaxis receptor. AB - The bacterial chemotaxis receptors are transmembrane receptors with a simple signalling pathway which has elements relevant to the general understanding of signal recognition and transduction across membranes, how signals are relayed between molecules in a pathway, and how adaptation to a persistent signal is achieved. In contrast to many mammalian receptors which signal by oligomerizing upon ligand binding, the chemotaxis receptors are dimeric even in the absence of their ligands, and their signalling does not depend on a monomer-dimer equilibrium. Bacterial chemotaxis receptors are composed of a ligand-binding domain, a transmembrane domain consisting of two helices TM1 and TM2, and a cytoplasmic domain. All known bacterial chemotaxis receptors have a highly conserved cytoplasmic domain, which unites signals from different ligand domains into a single signalling pathway to flagella motors. Here we report the crystal structure of the cytoplasmic domain of a serine chemotaxis receptor of Escherichia coli, which reveals a 200 A-long coiled-coil of two antiparallel helices connected by a 'U-turn'. Two of these domains form a long, supercoiled, four-helical bundle in the cytoplasmic portion of the receptor. PMID- 10466732 TI - Data explosion fuels search for drugs. PMID- 10466733 TI - Cerebral phosphorus metabolite abnormalities in opiate-dependent polydrug abusers in methadone maintenance. AB - This study evaluated cerebral phosphorus metabolites in opiate-dependent polydrug abusers in methadone maintenance therapy (MMT) and determined whether metabolite profiles differed based on treatment duration. Phosphorus magnetic resonance spectroscopy (31P-MRS) data were acquired with the ISIS volume localization method from a 50-mm thick axial brain slice through the orbitofrontal and occipital cortices. Study subjects included 15 MMT subjects, seven having undergone treatment for an average of 39 +/- 23 weeks (mean +/- S.D.) and eight having undergone treatment for 137 +/- 53 weeks, as well as an age matched comparison group (n = 16). The methadone dose administered on the study day averaged 70.5 +/- 17.1 mg and was statistically equivalent in short- and long term subgroups. MMT subjects (n = 15) differed from control subjects in percent phosphocreatine (%PCr) levels (-13%), and in both phosphomonoester (%PME, +13%) and phosphodiester (%PDE, +10%) levels, which likely reflect abnormalities in energy and phospholipid metabolism, respectively. There were no sex effects or group by sex interaction effects on these measures. In short-term MMT treatment subjects, abnormal %PCr (-18%), %PME (+20%) and %PDE (+17%) levels were found compared with control subjects. The only metabolite abnormality detected in long term MMT subjects was decreased %PCr (-9%), in spite of continued illicit drug abuse. From these data, we conclude that polydrug abusers in MMT have 31P-MRS results consistent with abnormal brain metabolism and phospholipid balance. The nearly normal metabolite profile in long-term MMT subjects suggests that prolonged MMT may be associated with improved neurochemistry. PMID- 10466734 TI - Dopamine transporter availability and depressive symptoms during alcohol withdrawal. AB - Alcohol-related temporary depressive symptoms are hypothesized to be related to dopaminergic dysfunction. The aim of this study was to investigate whether or not depressive symptoms correlate with reduced dopamine transporter (DAT) availability. We studied the DAT availability in 28 alcoholic subjects with beta CIT ([123-iodium]-2-betacarbomethoxy-3-beta-(4-iodophenyl)-tropa ne) single photon emission tomography (SPET) and found a reduction in DAT availability during withdrawal that subsequently showed a significant increase during sobriety. The relationship between DAT availability and Montgomery-Asberg Depression Rating scale scores, both during withdrawal and after sobriety, was assessed. The main finding was a statistically significant correlation between DAT variances and depressive symptom scores during both states. The findings indicate a possible dopaminergic etiology for depressive symptoms in alcohol withdrawal, which suggests that dopaminergic antidepressants might be beneficial in the treatment of alcohol withdrawal. PMID- 10466735 TI - Positive and negative symptoms and regional cerebral perfusion in antipsychotic naive schizophrenic patients: a high-resolution SPECT study. AB - In 17 antipsychotic-naive schizophrenic patients, the scores of positive and negative symptoms of schizophrenia were correlated with relative regional cerebral perfusion measured by 99m-Tc-ECD (ethyl cysteinate dimer) single photon emission computed tomography (SPECT). Scans were performed in subjects at rest. The negative symptom dimension was significantly correlated with a decreased level of perfusion in the left thalamic region. Other non-significant trends were also observed; the positive symptom dimension was related to decreased perfusion in the left temporal region and to increased perfusion in the right frontal region, while the negative symptom dimension was related to increased perfusion in the left frontal region. These findings suggest that the positive and negative symptoms of schizophrenia are related to dysfunctions in different regions of the brain and different lateralized patterns of dysfunction. PMID- 10466736 TI - Low resolution brain electromagnetic tomography (LORETA) functional imaging in acute, neuroleptic-naive, first-episode, productive schizophrenia. AB - Functional imaging of brain electrical activity was performed in nine acute, neuroleptic-naive, first-episode, productive patients with schizophrenia and 36 control subjects. Low-resolution electromagnetic tomography (LORETA, three dimensional images of cortical current density) was computed from 19-channel electroencephalographic (EEG) activity obtained under resting conditions, separately for the different EEG frequencies. Three patterns of activity were evident in the patients: (1) an anterior, near-bilateral excess of delta frequency activity; (2) an anterior-inferior deficit of theta frequency activity coupled with an anterior-inferior left-sided deficit of alpha-1 and alpha-2 frequency activity; and (3) a posterior-superior right-sided excess of beta-1, beta-2 and beta-3 frequency activity. Patients showed deviations from normal brain activity as evidenced by LORETA along an anterior-left-to-posterior-right spatial axis. The high temporal resolution of EEG makes it possible to specify the deviations not only as excess or deficit, but also as inhibitory, normal and excitatory. The patients showed a dis-coordinated brain functional state consisting of inhibited prefrontal/frontal areas and simultaneously overexcited right parietal areas, while left anterior, left temporal and left central areas lacked normal routine activity. Since all information processing is brain-state dependent, this dis-coordinated state must result in inadequate treatment of (externally or internally generated) information. PMID- 10466738 TI - Visual assessment of medical temporal lobe atrophy in demented and healthy control subjects: correlation with volumetry. AB - The present study evaluated the validity of visual rating of medial temporal lobe atrophy on coronal magnetic resonance imaging scans in a population of demented and non-demented individuals. Medial temporal lobe atrophy in 194 subjects was visually rated from hard copies, using a 0-4 rating scale, and a comparison was made with the absolute volumes (ccm) of the medial temporal lobe as estimated with volumetry, using a stereological method. We found a highly significant correlation between the estimated and stereologically measured volumes. There was a 10-fold difference in time spent on rating medial temporal lobe atrophy (1-2 min) vs. time spent calculating the medial temporal lobe volume (10-12 min) on a single subject. The diagnostic accuracy of both methods showed that visual rating was more efficient than volumetry in differentiating Alzheimer's disease from control subjects, We conclude that visual rating is a reliable and fast method to estimate medial temporal lobe atrophy in demented subjects in a clinical setting. PMID- 10466737 TI - Topography of callosal atrophy reflects distribution of regional cerebral volume reduction in Alzheimer's disease. AB - It has been suggested that regional corpus callosum atrophy in Alzheimer's disease (AD) may serve as an in vivo index of neuronal loss in the neocortex. In this study total and regional size of the corpus callosum was evaluated with respect to the volumes of the frontal, temporal, and parietal lobes in 38 patients with AD (NINCDS-ADRDA criteria) using quantitative magnetic resonance imaging. Twenty healthy subjects matched for age and gender served as a control group. All quantitative measurements were performed by manual tracing using personal computer-based software. Both total size and the five measured regional subsections were significantly smaller in AD when compared to the control subjects. The severity of dementia was significantly correlated with the size of the middle sections of the corpus callosum (rostral body and midbody). Within the AD group, the rostral body of the corpus callosum was significantly correlated with the frontal lobe volumes, the midbody was correlated with the temporal lobe volumes, and size of the splenium was correlated with the parietal lobe volumes. We conclude that callosal atrophy in AD reflects the severity and pattern of cortical neuronal damage. Correlations between regional callosal atrophy and severity of dementia indicate that interhemispheric cortico-cortical disconnection may contribute to the dementia syndrome. PMID- 10466739 TI - Identification of highly methylated arginine residues in an endogenous 20-kDa polypeptide in cancer cells. AB - Enzymatic methylation of endogenous proteins in several cancer cell lines was investigated to understand a possible relationship between protein-arginine methylation and cellular proliferation. Cytosolic extracts prepared from several cancer cells (HeLa, HCT-48, A549, and HepG2) and incubated with S-adenosyl-L [methyl-3H]methionine revealed an intensely [methyl-3H]-labeled 20-kDa polypeptide. On the other hand, cytosolic extracts prepared from normal colon cells did not show any methylation of the 20-kDa protein under identical conditions. To identify nature of the 20-kDa polypeptide, purified histones were methylated with HCT-48 cytosolic extracts and analyzed by SDS-PAGE. However, none of the histones comigrated with the methylated 20-kDa polypeptide, indicating that it is unlikely to be any of the histone subclasses. The [methyl-3H]group in the 20-kDa polypeptide was stable at pH 10-11 (37 degrees C for 30 min) and methylation was not stimulated by GTPgammaS (4 mM), thus the reaction is neither carboxyl methylesterification on isoaspartyl residues, nor on C-terminal farnesylated cysteine. The present study together with the previous identification of N(G)-methylated arginine residues in the HCT-48 cytosol fraction suggests that this novel endogenous 20-kDa arginine-methylation is a cellular proliferation-related posttranslational modification reaction. PMID- 10466740 TI - Chronic early leucine administration induces behavioral deficits in rats. AB - Sustained levels of leucine comparable to those of human Maple Syrup Urine Disease (MSUD) were achieved in blood and brain of rats by subcutaneous leucine administration twice a day from the 6th to the 28th day of life. Control rats were treated with saline in the same volumes. Behavioral studies using aversive and nonaversive tasks were performed during adult age. Chronic early leucine treatment impaired acquisition of a two-way shuttle avoidance task and altered habituation to an open field. Our results suggest that early postnatal leucine administration induces long-lasting behavioral deficits. PMID- 10466741 TI - Detection of bikunin mRNA in limited portions of rat brain. AB - Tissue distribution of bikunin mRNA, which encodes a Kunitz-type serine protease inhibitor of the inter-alpha-inhibitor family (IalphaI), was studied in rats and mice by the reverse-transcripsion polymerase chain reaction (RT-PCR). We found that the liver as well as other tissues, such as the kidney, testis and adrenal gland, expressed bikunin mRNA. Although signals of bikunin mRNA were faint in the whole brain of rats and mice, distinct signals were found in limited portions of rat brain, such as the hippocampus, cerebral cortex and pituitary, but undetectable in cerebellum, medulla oblongata, hypothalamus, striatum, midbrain and choroid plexus. In three distinct types of cells, such as neurons, astrocytes and meningeal cells, in primary cultures isolated from the cerebral cortex and meninges of 1-day-old newborn rats, only neurons positively expressed bikunin mRNA. These results suggest that, in addition to peripheral tissues, neurons in the hippocampus and cerebral cortex produce bikunin, suggesting a potential role of bikunin/IalphaI family in these brain regions. PMID- 10466742 TI - Effect of suplatast tosilate (IPD-1151T) on cytokine production by allergen specific human Th1 and Th2 cell lines. AB - Suplatast tosilate (IPD-1151T) is an antiallergic agent that suppresses airway eosinophil infiltration in asthma. We investigated the effects of IPD-1151T on proliferative response and cytokine production by human antigen-specific T cell lines. Purified protein derivatives (PPD)-specific T helper 1 (Th1) cell lines and Dermatophagoides farinae (Der f)-specific T helper 2 (Th2) cell lines were established from patients with asthma sensitized with house dust mite. Stimulation of PPD-specific and Der f-specific T cell lines with relevant antigens resulted in production of mostly interferon (IFN)-gamma and of interleukin (IL)-4 and IL-5, respectively. IPD-1151T did not inhibit the proliferative responses of either the Th1 or Th2 cell line to antigens. Although IPD-1151T did not inhibit IFN-gamma production by PPD-specific Th1 cell lines, it did inhibit IL-4 and IL-5 production by antigen-stimulated Der f-specific Th2 cell lines in a dose-dependent manner. IPD-1151T directly inhibited cytokine production by Der f-specific Th2 cell lines stimulated with immobilized anti-CD3 antibodies. Although IPD-1151T did not inhibit the clonal expansion of memory T cells among PBMCs into PPD-specific Th1 and Th2 cell lines, it did inhibit IL-4 and IL-5 production by Der f-specific Th2 cell lines but not IFN-gamma production by PPD-specific Th1 cell lines. These results suggest that IPD-1151T selectively inhibits Th2-type cytokine production. PMID- 10466743 TI - Effects of PAMP on mRNAs coding for catecholamine-synthesizing enzymes in PC12 cells. AB - Proadrenomedullin N-terminal 20 peptide (PAMP) is a novel hypotensive peptide found in the N-terminal portion of the precursor of adrenomedullin (AM). Although PAMP and AM originate from the same precursor and exert both a potent hypotensive action, they seem to control blood pressure through different mechanisms. To gain new insight into the anticholinergic actions of PAMP, we determined the effects of PAMP on the tyrosine hydroxylase (TH)- and dopamine beta-hydroxylase (DBH) mRNA expression in the rat pheochromocytoma cell line PC12 stimulated by nicotine. PAMP (> or =1 microM) significantly inhibited the nicotine-induced increases of TH- and DBH mRNA expression in a concentration-dependent manner. Also, PAMP at the concentrations (> or =1 microM) significantly inhibited nicotine-induced cyclic adenosine monophosphate (cAMP) production. These results indicate that the anticholinergic hypotensive actions of PAMP can be explained, at least in part, by its inhibition of the expression of mRNAs coding for catecholamine-synthesizing enzymes, and that the inhibitory effect is mediated by the cAMP/protein kinase A pathway. PMID- 10466744 TI - Different apolipoprotein B breakdown patterns in models of oxidized low density lipoprotein. AB - Low density lipoprotein (LDL) oxidation is characterized by alterations in biological properties and structure of the lipoprotein particles, including breakdown and modification of apolipoprotein B (apoB). We compared apoB breakdown patterns in different models of minimally and extensively oxidized LDL using Western blotting techniques and several monoclonal and polyclonal antibodies. It was found that copper and endothelial cell-mediated oxidation produced a relatively similar apoB banding pattern with progressive fragmentation of apoB during LDL oxidation, whereas malondialdehyde (MDA)- and hydroxynonenal (HNE) modified LDL produced an aggregated apoB. It is conceivable that apoB fragments present in copper and endothelial cell oxidized LDL lead to the exposure on the lipoprotein surface of different protein epitopes than in aggregated MDA-LDL and HNE-LDL. Although all models of extensively oxidized LDL led to increased lipid uptake in macrophages, mild degrees of oxidation interfered with LDL uptake in fibroblasts and extensively oxidized LDL impaired degradation of native LDL in fibroblasts. We suggest that in order to improve interpretation and comparison of results, data obtained with various models of oxidized LDL should be compared to the simpliest and most reproducible models of 3 h and 18 h copper-oxidized LDL (apoB breakdown) and MDA-LDL (apoB aggregation) since different models of oxidized LDL have significant differences in apoB breakdown and aggregation patterns which may affect immunological and biological properties of oxidized LDL. PMID- 10466745 TI - Endothelial cell chemotaxic activity expressed in rat placenta is not associated with prolactin-like proteins B and C. AB - Conditioned medium from gestation day 18 rat placental cultures showed potent stimulation of the directional migration of human retinal endothelial cells. To examine the role of major secreted placental proteins in this chemotaxic activity, prolactin-like proteins (PLPs)-B and C were purified from rat placenta using immuno-affinity chromatography. In contrast to conditioned medium, native PLP-B and PLP-C preparations failed to show any significant stimulation of endothelial cell migration. This study further examined the ability of PLP-B to bind to rat receptors for growth hormone (GH-R) and prolactin (PRL-R). In competitive binding assays with [125I]-hGH, neither native nor recombinant PLP-B preparations showed significant high affinity binding to the transfected rat GH-R or PRL-R. In summary, neither PLP-B nor PLP-C exhibit the potent chemotaxis stimulatory activity of placental conditioned media, nor does PLP-B show evidence of ability to act via rat GH or PRL receptors. PMID- 10466746 TI - Expression and localization of protein kinase C theta isoform in mouse testis. AB - Protein kinase C (PKC) is encoded by a complex of a gene family, and its multiple isoforms are expressed in various mammalian tissues. The objective of this study was to investigate the expression and localization of a PKC theta isoform in mouse testis. PKC theta displays the highest homology to PKC delta, lacks the Ca2+-binding C2 domain and, thus, belongs to the subfamily of Ca2+-independent PKC enzymes which also includes the delta, epsilon, zeta and eta isoforms. We analyzed the PKC theta mRNA and protein by Northern blotting, in situ hybridization, and immunohistochemistry. In testes of normal mice, signals of PKC theta isoform expression were detected specifically in the interstitial cells of testes. The expression of PKC theta isoform was also detected in testes of germ cell-deficient W/W(v) mice. These results suggest that PKC theta isoform has the specific biological functions in the interstitial cells of testis. PMID- 10466747 TI - Modulation of enteric cholinergic neurons by hetero- and autoreceptors: cooperation among inhibitory inputs. AB - In the guinea-pig colon, acetylcholine (ACh) release from intrinsic cholinergic motor neurons is inhibited by adrenoceptors, opioid and muscarinic receptors. Chronic sympathetic denervation resulted in supersensitivity to the inhibitory effect of DAMGO (mu-opioid agonist) on ACh release and on the peristaltic reflex. After chronic treatment with naltrexone (NTX) supersensitivity to DAMGO and subsensitivity to UK14,304 (alpha2-adrenoceptor agonist) developed for both functional parameters. The facilitatory effect of scopolamine on ACh release remained unchanged after chronic NTX treatment, whereas it was potentiated after chronic sympathetic denervation. These data suggest the existence of a functional interaction between different inhibitory pathways modulating cholinergic motor neurons in the guinea-pig colon. Namely, chronic manipulation of an inhibitory pathway may entail adaptive sensitivity changes in another inhibitory pathway so that homeostasis can be maintained. PMID- 10466748 TI - Influence of the cannabinoid agonist HU 210 on cocaine- and CQP 201-403-induced behavioural effects in rat. AB - Acute injection of the cannabinoid agonist HU 210 (6.25-100 microg/kg, i.p.) dose dependently inhibited rat locomotor activity and rearing, while subchronic treatment with the drug (once daily for 7 days) at the same doses only diminished locomotion. Acute but not subchronic administration of HU 210 (12.5-50 microg/kg, i.p.) potently counteracted acute and subchronic cocaine (15 mg/kg, i.p.)-induced hyperlocomotion and enhanced rearing. The acute cannabinoid (6.25-100 microg/kg, i.p.) also inhibited locomotor activity, stereotyped behaviour and shaking elicited by the D1/D2 agonist CQP 201-403 (500 microg/kg, i.p.). On the contrary, subchronic treatments with HU 210 enhanced CQP 201-403-induced locomotor activity and potently stimulated escape attempts. Discussion centers on the influence of cannabinoids on experimental models of psychosis. PMID- 10466749 TI - Effect of lysophosphatidic acid on the ovum transport in mouse oviducts. AB - The effects of lysophosphatidic acid (LPA) on ovum transport in mouse oviducts were studied. When excised oviducts were incubated at 37 degrees C under 5% CO2 in humidified air for 24 hours, addition of LPA at 10 microM to the medium significantly accelerated the rate of ovum transport, and 1 microM LPA slightly increased the ovum transport rate. These increases were not inhibited by 10 microM indomethacin, a cyclooxygense inhibitor, but were suppressed by 260 ng/ml of pertussis toxin or 10 microM verapamil, a voltage-sensitive calcium channel blocker. These data suggested that LPA stimulates mouse ovum transport by contracting oviductual smooth muscle via a voltage-sensitive calcium channel mediated by a pertussis toxin-sensitive G-protein-linked receptor. PMID- 10466750 TI - Coordinated distribution patterns of three enzyme activities involved in the absorption and metabolism of beta-carotene and vitamin A along the villus-crypt axis of chick duodenum. AB - The conversion of beta-carotene to retinal and the succeeding metabolic process of the retinal leading to production of retinol and retinyl esters are the prerequisite for the utilization of beta-carotene as a provitamin A. These processes are participated by beta-carotene cleavage enzyme, retinal reductase and retinol esterifying enzyme(s) in the small intestine. To examine whether these enzymes exhibit the coordinated distribution in the villus, we have used the cryostat sectioning technique to quantify the activities of beta-carotene cleavage enzyme, retinal reductase and retinol esterifying enzymes along the villus-crypt axis in 8-day-old chick duodenum. The beta-carotene cleavage enzyme activity was very low in the crypt and gradually increased, reaching a maximum in the mid-villus. The villus-crypt gradient of the beta-carotene cleavage enzyme activity corresponded with those of retinal reductase activity and lecithin: retinol acyltransferase (LRAT) activity, but distinct from that of acyl-CoA: retinol acyltransferase (ARAT) activity. Furthermore, the distribution of the content of retinyl esters was similar to that of LRAT activity. These results suggest that the beta-carotene cleavage enzyme is coordinately distributed along the villus-crypt axis with retinal reductase and LRAT, the two enzymes which require cellular retinol-binding protein, typeII (CRBPII) as the donor of the substrate. PMID- 10466751 TI - The many consequences of chemical- and genetic-based modulation of drug metabolizing enzyme activities. AB - The induction or inhibition of the metabolizing enzyme activities by a great deal of substances (including drugs) influence their toxicological or pharmacological outcomes as well as that of other xenobiotics or drugs to which human is simultaneously exposed. The dual bioactivating/detoxificating nature of both phase I and phase II enzymes poses such modulation as an unavoidable unhealthy phenomenon. Therefore, the proposed strategies in preventive medicine which foresee boosting or depressing enzymatic effects such as those in the field of cancer chemoprevention, should be carefully reconsidered before their credibility would be compromised. As the phenotypic features, genetic polymorphisms leading to the occurrence of high or low metabolizers in the population, each at high risk to certain forms of toxicity, behave as a sort of "constitutive" enzymatic modulation. Thus, considering the double-edged sword nature (detoxi-toxicant) of these catalysts towards ubiquitous environmental pollutants, the search for individual susceptibility by means of the genotypic analysis represents a very intriguing problem. However, the knowledge of the "overall" metabolic fingerprint associated to the phenotypic analysis in a single person could offer an interesting way to (partially) control human risk by making suitable (well aimed) modifications of determined life-styles (e.g. stop smoking or drinking) or particular dietetic practices (e.g. stop eating high cooked meat or fish) as well as selecting personalised drug adjustments by physicians either in terms of dosage or fitting drug. PMID- 10466753 TI - Exploiting the p53 pathway for cancer diagnosis and therapy. PMID- 10466752 TI - Enhanced cytotoxicity and suppression of glucose transport rate by combined treatment of recombinant human tumour necrosis factor-alpha and hyperthermia on L929 cells. AB - Combined treatment with human recombinant TNF-alpha (rhTNF-alpha) and hyperthermia at 43 degrees C arrested the growth of mouse fibrosarcoma L929 cells in vitro. The cytotoxic effect was enhanced in combined treatment compared with that following administration of rhTNF-alpha or hyperthermia alone. When the cells were subjected to hyperthermia at 43 degrees C for 3 hours and then incubated with 0.4 ng/ml rhTNF-alpha at 37 degrees C for 24 hours, a statistically significant 65% decrease in the rate of cellular glucose uptake was observed. This suppressive effect was synergistic in terms of effect achieved by rhTNF-alpha or hyperthermia individually. Since the growth of tumour cells depends mainly on catabolism of glucose, our findings indicate that one manner by which combined rhTNF-alpha and hyperthermia treatment inhibits L929 cell growth may be by reducing the supply of glucose to the cells. PMID- 10466754 TI - Cell cycle control and the mitotic spindle. PMID- 10466755 TI - DNA-dependent protein kinase gets a break: its role in repairing DNA and maintaining genomic integrity. PMID- 10466756 TI - Clinical applications of a novel mammalian cell-free DNA replication system. PMID- 10466757 TI - Signal transduction pathways regulated by the Rho family of small GTPases. PMID- 10466758 TI - Target genes and target cells in carcinogenesis. PMID- 10466759 TI - Apoptosis and carcinogenesis. AB - Apoptosis has long been known to be effected through a common sequence of structural changes, despite the wide variety of initiating stimuli. These common structural events appear to depend upon activation of a set of enzymes (caspases) which direct a strongly conserved, terminal effector pathway. The regulation of this pathway, and in particular its coupling to DNA damage, appears to be critical in maintaining at low levels the number of mutated cells within tissues. The frequency with which tumours (experimental and human) bear deficiency in p53 or MSH-2 repair function may indicate the importance of these proteins in coupling DNA damage to apoptosis. PMID- 10466760 TI - The cell cycle-regulating transcription factors E2F-RB. PMID- 10466761 TI - Characterization of a multi-tissue tumour suppressor and senescence gene. PMID- 10466762 TI - Costs, benefits and limitations of genetic testing for cancer risk. AB - There has been considerable progress in the identification of predisposing genes involved in many of the rare familial cancer syndromes and in a proportion of the common cancers. Genetic testing for cancer risk is now part of good medical practice in a number of cases, allowing specific cancer screening and prevention to be targeted to those with the greatest need. In some situations, the advantages of testing are less obvious, and careful evaluation of the costs and benefits of testing, and the efficacy of any subsequent intervention, is essential. In the meantime, it is clear that structures must be set up to evaluate demand for testing and deal with it; to inform and educate the public and doctors so that demand is based on understanding of what genetic tests can do; to provide action on test results and to advise and support individuals before and after testing. PMID- 10466763 TI - Targeting human tumours with antigen-specific cytotoxic T-cells. PMID- 10466764 TI - Tumour vasculature as a target for cancer therapy. PMID- 10466765 TI - Nucleic acid targeting: therapeutic strategies for the 21st century. PMID- 10466766 TI - Monitoring cancer by magnetic resonance. PMID- 10466767 TI - EPIC-Norfolk: study design and characteristics of the cohort. European Prospective Investigation of Cancer. PMID- 10466768 TI - Targeting cancer therapy. PMID- 10466769 TI - CHART (continuous, hyperfractionated, accelerated radiotherapy): a tale of two disciplines. PMID- 10466770 TI - New drug development: its role in reversing drug resistance. PMID- 10466771 TI - Pet Travel Scheme: further details from MAFF. PMID- 10466772 TI - Modelling the expected numbers of preclinical and clinical cases of bovine spongiform encephalopathy in Switzerland. AB - The objective of this study was to model the expected numbers of cattle incubating bovine spongiform encephalopathy (BSE) and the numbers of clinical cases of BSE in the Swiss cattle population between 1984 and 2005. The results were compared with the observed number of clinical BSE cases and with the results of a culling and testing scheme on herdmates of cattle with BSE. The age distribution of the Swiss cattle population, the age-at-death distribution of the first 235 BSE cases and exposure information were used to calculate the expected number of infected cattle in each birth cohort and the resulting numbers of clinical cases and survivors incubating the disease for each year. The model which did not assume any under-reporting of cases fitted the observed epidemic curve of clinical cases reasonably well, and predicted that the Swiss BSE epidemic would come to an end between 2003 and 2005. The age of survivors incubating BSE is increasing. The higher than expected incidence of subclinical cases observed in animals from the culling scheme is most probably the result of the heterogeneous distribution of infected animals and affected herds in the population. The results of the model need to be taken into account when designing surveillance and testing schemes for BSE. PMID- 10466773 TI - Incidence of foot and skin lesions in nursing piglets and their association with behavioural activities. AB - A total of 356 piglets from one farm were examined for foot and skin lesions every day for the first 10 days after birth, and then on alternate days until they were weaned. Over a period of 24 days 100 per cent of the piglets examined developed sole bruising, and 49.1 per cent developed sole erosions. Sole bruising lasted for an average of 13 days and sole erosions for seven days. At the beginning of the study, a higher proportion of piglets had mild sole bruising; from three to nine days of age piglets had moderate sole bruising but from days 10 to 20 mild bruising was again observed more frequently. Skin lesions were observed on the carpal aspect of the front limbs; 60.9 per cent of the piglets developed skin abrasions, 70.7 per cent developed healed wounds and 90 per cent developed hairless patches. The skin abrasions lasted on average for six days, the healed wounds for five days and the hairless patches for eight days. Piglets which developed sole bruising and/or sole erosions on the first day of life were significantly heavier than those which did not. Continuous observations of the piglets' behaviour during the first six days of life showed that sole bruising increased as the total time spent in the creep area or lying near the sow on the solid floor increased. During the first three days of life skin abrasions increased as the total time spent lying in the creep area increased. There was a positive correlation between the total time piglets spent suckling and the incidence of carpal skin abrasions between four and six days of age. Piglets with sole bruising, sole erosions or carpal skin abrasions spent less time during the day in 'other' activities such as walking, playing or fighting. PMID- 10466774 TI - Bullous systemic lupus erythematosus (type I) in a dog. AB - In human patients with systemic lupus erythematosus, cutaneous subepidermal blistering can occur because of the production of antibodies specific for basement membrane antigens. This condition is referred to as bullous systemic lupus erythematosus (BSLE). A dog was diagnosed with BSLE because it fulfilled the following criteria: (i) a diagnosis of systemic lupus erythematosus by standard methods; (ii) an acquired, vesicular, erosive and ulcerative eruption; (iii) microscopical subepidermal vesicles with neutrophil-predominant inflammation at the dermo-epidermal junction; (iv) deposition of IgG at the epidermal basement membrane zone; and (v) circulating IgG autoantibodies against type VII collagen. Anti-collagen VII type I-BSLE therefore needs to be considered as a possible differential diagnosis for canine autoimmune subepidermal blistering diseases. PMID- 10466775 TI - Presence of 3-nitropropionic acid, in widely distributed pasture legumes in Britain. PMID- 10466776 TI - Recovery of Microsporum gypseum and Malassezia pachydermatis from the nasal bridge in various dog groups. PMID- 10466777 TI - Control of sheep ectoparasites using shower dips, spray races and jetting wands. PMID- 10466778 TI - Pet travel scheme. PMID- 10466779 TI - Ragwort poisoning. PMID- 10466780 TI - Society of veterinary assistants? PMID- 10466781 TI - Retention fees and the MRCVS diploma. PMID- 10466782 TI - Balance of the sexes. PMID- 10466783 TI - Virus-encoded proteinases of the Togaviridae. PMID- 10466784 TI - 'Primer alignment-and-extension': a novel mechanism of viral RNA recombination responsible for the rescue of inactivated poliovirus cDNA clones. AB - In the course of experiments designed to assess the potential role of alternative open reading frames (ORF) present in the 5'-terminal untranslated region (5'-UTR) of poliovirus type 1 (Mahoney strain) genomic RNA, we came across a double mutation that completely abrogated the infectivity of full-length cDNA clones. The infectivity was rescued in trans by cotransfecting COS-1 cells with short RNA transcripts of the wild-type 5'-UTR of poliovirus type 2 Lansing, provided a free 3'-OH was available. Direct sequencing of the viral RNA revealed that the infectious viruses recovered were recombinants Lansing/Mahoney, with variable points of 'crossing-over'. A novel mechanism of RNA-RNA recombination, which we propose to call 'primer alignment-and-extension', is described that would explain the high rate of recombination of RNA viruses observed in natural conditions. PMID- 10466785 TI - Antigenic properties and population stability of a foot-and-mouth disease virus with an altered Arg-Gly-Asp receptor-recognition motif. AB - The antigenic properties and genetic stability of a multiply passaged foot-and mouth disease virus (FMDV) clone C-S8c1 with an Arg-Gly-Gly triplet (RGG) instead of the Arg-Gly-Asp (RGD) integrin-recognition motif at positions 141 to 143 of capsid protein VP1 are described. Clear antigenic differences between FMDV RGG and clone C-S8c1 have been documented in ELISA, enzyme-linked immunoelectrotransfer (Western) blot and neutralization assays using site A specific monoclonal antibodies and anti-FMDV polyclonal antibodies from swine and guinea pigs. The results validate with a live virus the role of the RGD (in particular Asp-143) in recognition of (and neutralization by) antibodies, a role previously suggested by immunochemical and structural studies with synthetic peptides. The FMDV RGG was genetically stable in a large proportion of serial infections of BHK-21 cells. However, a revertant virus with RGD was generated in one out of six passage series. Interestingly, this revertant FMDV did not reach dominance but established an equilibrium with its parental FMDV RGG, accompanied by an increase of quasispecies complexity at the sequences around the RGG triplet. FMDV RGG exhibited a selective disadvantage relative to other RGD containing clones isolated from the same parental FMDV population. The results suggest that large antigenic variations can be prompted by replacements at critical capsid sites, including those involved in receptor recognition. These critical replacements may yield viruses whose stability allows them to replicate efficiently and to expand the sequence repertoire of an antigenic site. PMID- 10466786 TI - Evidence for the role of His-142 of protein 1C in the acid-induced disassembly of foot-and-mouth disease virus capsids. AB - Foot-and-mouth disease virus (FMDV) capsids are inherently labile under mildly acidic conditions, dissociating to pentamers at pH values in the region of 6.5, with the release of protein 1A and the viral RNA. This acid-induced disassembly is thought to be required for the entry of the virus genome into the host cell. Previous work has highlighted a histidine-alpha-helix charge-dipole interaction at the twofold axes of symmetry between pentamers and has suggested that this interaction plays a role in acid-induced disassembly. The validity of this theory has now been tested by converting the implicated residue, His-142 of protein 1C, to Arg, Phe and Asp. The effects of such changes were studied by using a previously described vaccinia virus expression system, in which synthesis and processing of FMDV capsid proteins results in the self-assembly of capsids. In agreement with the histidine-alpha-helix charge-dipole theory, assembly in the arginine mutant was found to be greatly reduced, while capsids of the aspartic acid mutant were considerably more stable under acidic conditions than the wild type. Aberrant but acid-stable complexes were obtained in the phenylalanine mutant. PMID- 10466787 TI - Site-saturation mutagenesis of the PALTAVETG motif in coxsackievirus A9 capsid protein VP1 reveals evidence of conservation of a periodic hydrophobicity profile. AB - Enteroviruses possess a highly conserved 9 amino acid stretch of mainly hydrophobic character in the capsid protein VP1. A novel strategy, combining site saturation mutagenesis and a single-tube cloning and transfection procedure, has been developed for the analysis of this motif in coxsackievirus A9 (CAV-9). Four individual amino acids were separately mutated. Mutagenesis of three of the four positions in CAV-9 resulted in a number of viable but impaired mutant strains, each containing a single amino acid substitution. In contrast, no mutants with amino acid substitutions at leucine 31 were isolated, although three different leucine codons were found among the viruses recovered. Small plaque size was regularly associated with reduced yields of infectious virus and an amino acid substitution at the target site in the viruses isolated from the site-saturated virus pools. From the range of amino acids observed in viable mutants, it was possible to estimate the characteristics that are required at individual amino acid positions. It seems that in the motif studied here, a periodic hydrophobicity profile needs to be conserved. The constraints observed on the ranges of acceptable amino acids presumably reflect the structural-functional requirements that have resulted in the conservation of the motif. PMID- 10466788 TI - Sequence analysis of a porcine enterovirus serotype 1 isolate: relationships with other picornaviruses. AB - The majority of the genomic sequence of a porcine enterovirus serotype 1 (PEV-1) isolate was determined. The genome was found to contain a large open reading frame which encoded a leader protein prior to the capsid protein region. This showed no sequence identity to other picornavirus leader regions and the sequence data suggested that it does not possess proteolytic activity. The 2A protease was small and showed considerable sequence identity to the aphthoviruses and to equine rhinovirus serotype 2. The 2A/2B junction possessed the typical cleavage site (NPG/P) exhibited by these viruses. The other proteins shared less than 40% sequence identity with equivalent proteins from other picornavirus genera. Phylogenetic analyses of the P1 and 3D sequences indicated that this virus forms a distinct branch of the family Picornaviridae. On the basis of results presented in this paper PEV-1 has been assigned to a new picornavirus genus. The phylogeny of the virus in relation to other picornaviruses is discussed. PMID- 10466789 TI - The C-terminal region of the hepatitis C virus E1 glycoprotein confers localization within the endoplasmic reticulum. AB - Expression of the hepatitis C virus glycoprotein E1 in cultured cells localizes it to the endoplasmic reticulum, suggesting that E1 contains a signal mediating retention. Fusion of the C-terminal region of E1 to the ectodomain of CD4 prevented it from being transported to the cell surface. Fusion of this region of E1 resulted in localization of CD4 and influenza virus haemagglutinin chimeric molecules to a pre-medial Golgi compartment. This signal was present within E1 residues 311-383. Retention was not due to misfolding since the chimeric molecules did not form disulphide-linked aggregates indicative of misfolded proteins, and could be recognized by MAbs specific for conformational epitopes. PMID- 10466790 TI - Genetic stability of equine arteritis virus during horizontal and vertical transmission in an outbreak of equine viral arteritis. AB - An imported carrier stallion (A) from Europe was implicated in causing an extensive outbreak of equine viral arteritis (EVA) on a Warmblood breeding farm in Pennsylvania, USA. Strains of equine arteritis virus (EAV) present in the semen of two carrier stallions (A and G) on the farm were compared to those in tissues of foals born during the outbreak, as well as viruses present in the semen of two other stallions that became persistently infected carriers of EAV following infection during the outbreak. The 2822 bp segment encompassing ORFs 2 7 (nt 9807-12628; which encode the G(S), GP3, GP4, G(L), M and N proteins, respectively) was directly amplified by RT-PCR from semen samples and foal tissues. Nucleotide and phylogenetic analyses confirmed that virus present in the semen of stallion A initiated the outbreak. The genomes of viruses present in most foal tissues (10/11) and serum from an acutely infected mare collected during the outbreak were identical to that of virus present in the lung of the first foal that died of EVA. Virus in the placenta of one foal differed by one nucleotide (99.9% identity) from the predominant outbreak virus. The relative genetic stability of viruses that circulated during the outbreak contrasts markedly with the heterogeneous virus populations variously present in the semen of persistently infected stallions on the farm. These findings are consistent with the hypothesis that the carrier stallion can be a source of genetic diversity of EAV, and that outbreaks of EVA can be initiated by the horizontal aerosol transmission of specific viral variants that occur in the semen of particular carrier stallions. PMID- 10466791 TI - A novel internal open reading frame product expressed from a polycistronic mRNA of porcine epidemic diarrhoea virus may not contribute to virus attenuation. AB - Cell-culture-adapted (ca) porcine epidemic diarrhoea virus (PEDV) contains three internal open reading frames (I ORF) within the nucleocapsid protein gene and lacks the downstream counterpart of porcine transmissible gastroenteritis virus ORF7 or feline infectious peritonitis virus ORF6a. To confirm whether such features also exist in wild-type (wt) PEDV, the 3' 1800 nucleotides of its genome were sequenced and were found to be identical to those of ca virus. The coding potential of I-1 ORF was ascertained by transient expression in Vero cells followed by immunofluorescence using antipeptide sera. The I-1 protein was synthesized as a 12 kDa non-phosphorylated PEDV-specific protein that was not present in detectable amounts in virions. However, a low copy number of I-1 in the virion would suggest it is a structural component. Nevertheless, identical nucleotide sequences and gene expression strategies of attenuated ca virus and its virulent parent, wt PEDV, demonstrate that the 3' 1800 nucleotides or the genes and gene products encoded therein may not contribute to virus attenuation. PMID- 10466792 TI - Co-expression of a trans-dominant negative mutant of the human immunodeficiency virus type 1 (HIV-1) Rev protein affects the Rev-dependent splicing pattern and expression of HIV-1 RNAs. AB - Trans-dominant negative mutants of the human immunodeficiency virus type 1 (HIV 1) regulatory protein Rev inhibit the function of wild-type Rev in a dose dependent manner. This was previously shown to be caused by nuclear retention of the wild-type protein. In the present work, further analysis of the trans dominant negative effect was performed using cotransfection experiments with different constructs encoding HIV-1 Rev and viral structural proteins together with a plasmid encoding a trans-dominant negative Rev mutant. Thus, one species of pre-mRNA was transcribed from the reporter plasmids. This pre-mRNA was then either spliced or exported by Rev as unspliced RNA for translation of the HIV structural proteins. An immunofluorescence assay and Western blot analysis were used for analysis of protein expression. In situ hybridization was applied for labelling of unspliced mRNA in transfected cells, and RNase protection analysis was used to determine the relative amount of unspliced versus spliced mRNAs. The experiments confirmed that the transdominant negative mutant inhibited nuclear export of unspliced mRNA. It was, in addition, demonstrated for the first time that the trans-dominant negative mutant also affected a Rev-dependent regulatory step connected with viral pre-mRNA splicing. As a consequence, proteins expressed from unspliced and singly spliced HIV mRNAs decreased while there was an increase in protein products encoded by spliced and alternatively spliced mRNAs. PMID- 10466793 TI - Mutations in the env gene of human immunodeficiency virus type 1 NDK isolates and the use of African green monkey CXCR4 as a co-receptor in COS-7 cells. AB - A previous report from this laboratory described the isolation of the first CD4 independent human immunodeficiency virus type 1 isolate, m7NDK. This independence of CD4 is due to seven mutations located in the C2, V3 and C3 regions of the gp120 protein. The present report describes the entry features of the m5NDK virus, which contains five of the seven m7NDK mutations, located in the V3 loop and C3 region. The entry of this virus is strictly CD4-dependent but it can fuse with African green monkey (agm) COS-7 cells bearing human CD4 (h-CD4). This fusion is directly due to the five mutations in the envgene. It has also been shown that entry of m7NDK is CD4-independent in COS-7 cells. Since the wild-type NDK and m7NDK viruses use the human CXCR4 protein as co-receptor, agm-CXCR4 was cloned and used in transfection and fusion inhibition experiments to show that this receptor can be used by the m5 and m7NDK viruses. The wild-type NDK virus, which does not enter COS-7 cells, can use agm-CXCR4, but only when the receptor is transfected into target cells. Although co-receptor nature and expression levels are still major determinants of virus entry, this is the first case where a few mutations in the env gene can overcome this restriction. PMID- 10466794 TI - Manganese cations increase the mutation rate of human immunodeficiency virus type 1 ex vivo. AB - Human immunodeficiency virus (HIV) reverse transcription is an error-prone process with an overall mutation rate of approximately 3.4 x 10(-5) per base per replication cycle. This rate can be modulated by changes in different components of the retrotranscription reaction. In particular, in vitro substitution of magnesium cations (Mg2+) by manganese cations (Mn2+) has been shown to increase misincorporation of deoxynucleotide triphosphates (dNTPs) and to alter substrate specificity. Here, it is shown that Mn2+ also increases the HIV mutation rate ex vivo. Treatment of permissive cells with Mn2+ and subsequent HIV infection resulted in at least 6-fold and 10-fold increases in the mutant and mutation frequencies respectively, thus illustrating a further example of how to influence HIV genetic variation. PMID- 10466795 TI - Human neutralizing human immunodeficiency virus type 2-specific Fab molecules generated by phage display. AB - A panel of human immunodeficiency virus type 2 (HIV-2)-neutralizing, recombinant Fab fragments was generated by using the phage display technique. The combinatorial library was derived from an asymptomatic, HIV-2-seropositive individual and constructed on the surface of filamentous phage by using the pComb3 phagemid vector and then screened against native HIV-2 envelope glycoprotein (gp125). Ten of 30 Fab fragments generated displayed strong reactivity in an ELISA and were therefore selected for further study. Six of these possessed neutralizing capacity, with titres varying from 20 to 80 against the homologous HIV-2 strain, and one also had a weak neutralizing capacity against a heterologous HIV-2 isolate, K135. Sequencing of the heavy chain CDR3 regions showed that the gp125-specific Fabs represented individual clones. These reagents may be useful for studies on the conformational structures of the HIV-2 envelope antigens and their immunogenicity, which may help in vaccine design. Furthermore, the cloned Fab genes may be transformed into whole IgG for eukaryotic expression, and as such used for therapeutic and immunoprophylactic studies in HIV-2-infected macaques and, possibly, for human immunoprophylaxis against HIV-2. PMID- 10466796 TI - Identification and phylogenetic characterization of a human T-cell leukaemia virus type I isolate from a native inhabitant (Rapa Nui) of Easter Island. AB - Human T-cell leukaemia virus type I (HTLV-I) is endemic in Melanesia, one of the three ethnogeographic regions of the Pacific; in the other two regions, Polynesia and Micronesia, the incidence of the virus is relatively low. In an effort to gain new insights into the prevalence of HTLV-I in the Pacific region, we did a seroepidemiological survey on Easter Island, which is located on the eastern edge of Polynesia. Of 138 subjects surveyed, including 108 Rapa Nui (the native inhabitants of this island), we identified one HTLV-I-seropositive Rapa Nui. The new HTLV-I isolate derived from this carrier (E-12) was phylogenetically analysed to ascertain the origin and past dissemination of HTLV-I in the island. The analysis demonstrated that isolate E-12 belongs to subgroup A of the Cosmopolitan group, and that it differs from HTLV-Is found in Melanesia, which are highly divergent variants. In subgroup A, E-12 grouped with South American HTLV-Is including those from Amerindians. This result suggests that this isolate originated in South America rather than in Melanesia. PMID- 10466797 TI - Properties of human foamy virus relevant to its development as a vector for gene therapy. AB - The Spumaviridae (foamy viruses) are increasingly being considered as potential vectors for gene therapy, yet little has been documented of their basic cell biology. This study demonstrates that human foamy virus (HFV) has a broad tropism and that the receptor for HFV is expressed not only on many mammalian, but on avian and reptilian cells. Receptor interference assays using an envelope expressing cell line and a vesicular stomatitis virus/HFV pseudotype virus demonstrate that the cellular receptor is common to all primate members of the genus. The majority of foamy virus particles assemble and remain sequestered intracellularly. A rapid and quantitative method of assaying foamy virus infectivity by reverse transcriptase activity facilitates the use of classical protocols to increase infectious virus titres in vitro to > or = 10(6) TCID/ml. PMID- 10466798 TI - Detection and characterization of proteins encoded by the second ORF of the M2 gene of pneumoviruses. AB - The nucleotide sequence of the M2 gene of pneumonia virus of mice (PVM) was determined. The sequence showed that the gene encoded a protein of 176 amino acids with a predicted molecular mass of 20165 Da from a major ORF, which is smaller than the equivalent proteins encoded by human, bovine and ovine respiratory syncytial (RS) viruses. The PVM M2 protein is conserved, having 41% similarity to the equivalent human RS virus protein. In common with the M2 genes of the RS viruses and avian pneumovirus (APV), the PVM mRNA also contained a second ORF (ORF2) that partially overlaps the first ORF and which is capable of encoding a 98 residue polypeptide. No significant sequence identity could be detected between the putative M2 ORF2 proteins of PVM, APV and the RS viruses. The expression of the M2 ORF2 proteins of the pneumoviruses was investigated by using monospecific antisera raised against GST fusion proteins. Western blot analysis demonstrated the presence of polypeptides encoded by M2 ORF2 of PVM and RS virus corresponding with those predicted by in vitro translation studies, but this was not the case for APV. The PVM polypeptide was present as three distinct products in vivo. The PVM and RS virus polypeptides were also detected in cells by immunofluorescence, which showed that both were present in the cytoplasm with a degree of localization in inclusion bodies. No APV M2 ORF2 protein could be detected in vivo. The RS virus M2 ORF2 polypeptide was shown to accumulate during infection and the potential implications of this are discussed. PMID- 10466799 TI - Mapping of domains on the human parainfluenza virus type 2 nucleocapsid protein (NP) required for NP-phosphoprotein or NP-NP interaction. AB - The epitopes recognized by 41 monoclonal antibodies directed against the nucleocapsid protein (NP) of human parainfluenza virus type 2 (hPIV-2) were mapped on the primary structure of the hPIV-2 NP protein by testing their reactivities with deletion mutants. By Western immunoblotting using these monoclonal antibodies, the analysis of deletion mutants of the hPIV-2 NP protein was performed to identify the region essential for NP-NP interaction and phosphoprotein (P)-binding sites on the NP protein. The results indicate that the N-terminal 294 aa of the NP protein are all required for NP-NP self-assembly, and that two C-terminal parts of the NP protein are essential for NP-P binding: one region, aa 295-402, is required for binding to the C-terminal part of the P protein and another region, aa 403-494, to the N-terminal part of the P protein. PMID- 10466800 TI - Measles virus-induced immunosuppression in cotton rats is associated with cell cycle retardation in uninfected lymphocytes. AB - Measles virus (MV)-induced immune suppression during acute measles often leads to secondary viral, bacterial and parasitic infections which severely complicate the course of disease. Previously, we have shown that cotton rats are a good animal model to study MV-induced immune suppression, where proliferation inhibition after ex vivo stimulation of cotton rat spleen cells is induced by the viral glycoproteins (fusion and haemagglutinin proteins). We have now tested a variety of putative mechanisms of MV-induced immune suppression in this animal model. Proliferation inhibition is not due to fusion mediated by the MV glycoproteins and subsequent lysis of cells. Other putative mechanisms like classical anergy (unresponsiveness towards IL-2) or apoptosis do not seem to play a role in MV induced immune suppression. In contrast, it was shown that spleen cells from infected animals preferentially accumulate in the G0/G1 phase and progress more slowly through the cell cycle after mitogen stimulation in comparison to cells from non-infected animals. These data indicate a retardation of the cell cycle which is correlated with proliferation inhibition and might have severe consequences in mounting an effective immune response. PMID- 10466801 TI - Expression in cattle of epitopes of a heterologous virus using a recombinant rinderpest virus. AB - We have investigated the bovine immune response to heterologous proteins expressed using a recombinant rinderpest virus (RPV). A new gene unit was created in a cDNA copy of the genome of the vaccine strain of RPV, and an open reading frame inserted that encodes the polymerase (3Dpol) and parts of the capsid protein VP1 from foot-and-mouth disease virus (FMDV). Infectious recombinant RPV was rescued and shown to express the FMDV-derived protein at good levels in infected cells. The rescued virus was only slightly more attenuated in tissue culture than the original virus. Cattle infected with this recombinant generated a normal immune response to RPV, and were protected from lethal challenge by that virus. Experimental animals showed a specific delayed-type hypersensitivity response to FMDV 3Dpol, similar to that seen in FMDV infection; however, no antibodies were detected recognizing either of the components of the FMDV-derived protein, nor was any proliferative response to these epitopes found in isolated peripheral blood lymphocytes from infected animals. No protection was seen against FMDV infection. PMID- 10466802 TI - Dynamics of rabies virus quasispecies during serial passages in heterologous hosts. AB - To understand the mutations and genetic rearrangements that allow rabies virus infections of new hosts and adaptation in nature, the quasispecies structure of the nucleoprotein and glycoprotein genes as well as two noncoding sequences of a rabies virus genome were determined. Gene sequences were obtained from the brain and from the salivary glands of the original host, a naturally infected European fox, and after serial passages in mice, dogs, cats and cell culture. A relative genetic stasis of the consensus sequences confirmed previous results about the stability of rabies virus. At the quasispecies level, the mutation frequency varies, in the following order: glycoprotein region (21.9 x 10(-4) mutations per bp), noncoding sequence nucleoprotein-phosphoprotein region (7.2-7.9 x 10(-4) mutations per bp) and nucleoprotein gene region (2.9-3.7 x 10(-4) mutations per bp). These frequencies varied according to the number, type of heterologous passages and the genomic region considered. The shape of the quasispecies structure was dramatically modified by passages in mice, in which the mutation frequencies increased by 12-31 x 10(-4) mutations per bp, depending on the region considered. Non-synonymous mutations were preponderant particularly in the glycoprotein gene, stressing the importance of positive selection in the maintenance and fixation of substitutions. Two mechanisms of genomic evolution of the rabies virus quasispecies, while adapting to environmental changes, have been identified: a limited accumulation of mutations with no replacement of the original master sequence and a less frequent but rapid selective overgrowth of favoured variants. PMID- 10466803 TI - Effect of population patchiness and migration rates on the adaptation and divergence of vesicular stomatitis virus quasispecies populations. AB - The effect of migration among different isolated virus quasispecies populations on their adaptation and diversity was analysed through experimental evolution. An in vitro cell system was employed to simulate migration of vesicular stomatitis virus between isolated homogeneous host cell populations. The results clearly demonstrated a positive correlation between the migration rate and the magnitude of the mean fitness reached by the virus quasispecies populations. The results also showed, although less clearly, that fitness differences among quasispecies decreased with the magnitude of migration. These results are in close agreement with predictions of standard population genetics theory. These results can be explained in terms of the spread of beneficial mutations, originating in a single isolated quasispecies, through the entire system formed by the different quasispecies populations contained in different host cell populations. PMID- 10466804 TI - Characteristics of a new birnavirus associated with a warm-water fish cell line. AB - A warm-water fish cell line developed from blotched snakehead caudal peduncle (BSN) was found to have persistent birnavirus infection. Purified virus particles were of icosahedral shape and had 57+/-1.6 nm diameter. The BSN virus was resistant to 5-iodo-2'-deoxyuridine and induced yellowish-green cytoplasmic inclusions when stained with acridine orange. The virus was resistant to chloroform, acid and alkaline pH and heat treatment at 56 degrees C for 2 h. Purified virions had a buoyant density of 1.33 g/ml in CsCl and contained two genomic segments with molecular masses of 2.56 x 10(6) and 2.00 x 10(6) Da and four structural polypeptides of 112 (polyprotein, PP), 91 (VP1), 44 (VP2) and 37 (VP3) kDa. Reciprocal beta cross-neutralization tests incorporating four classical strains of infectious pancreatic necrosis virus (IPNV) (WB, Sp, Ab and TV-1) and the BSN virus established the complete serological distinctness of the virus from IPNV. Considering the uniqueness of the virus, the name blotched snakehead virus is proposed for this agent. PMID- 10466805 TI - Tissue culture infectivity of different strains of infectious bursal disease virus is determined by distinct amino acids in VP2. AB - Two types of strains of serotype I of infectious bursal disease virus (IBDV) have been described, on the basis of their ability (IBDV-TC) or inability (IBDV-BU) to infect chicken embryonic cells in culture. However, both types infect B lymphocytes in the bursa of Fabricius of young chickens. To determine the molecular basis for tissue culture infectivity, virus recombinants with chimeric segments A were constructed from IBDV-TC and IBDV-BU by reverse genetics. The region responsible for the different phenotypes was located in VP2. Site-directed mutagenesis identified single amino acids that are responsible for the restriction in infectivity. However, the appropriate amino acid exchanges are strain-specific. PMID- 10466806 TI - Sequence analysis and in vitro expression of genes 6 and 11 of an ovine group B rotavirus isolate, KB63: evidence for a non-defective, C-terminally truncated NSP1 and a phosphorylated NSP5. AB - An ovine group B rotavirus (GBR) isolate, KB63, was isolated from faeces of a young goat with diarrhoea in Xinjiang, People's Republic of China. Sequence determination and comparison of genes 6 and 11 with the corresponding sequences of GBR strains ADRV and IDIR showed that they were the cognate genes encoding NSP1 and NSP5, respectively. While the overall identities of nucleotide sequences between these two genes and the corresponding genes of strains ADRV and IDIR were in the range 52.6-57.2%, the identities of deduced amino acid sequences were only 34.9-46.3%. These results demonstrate that the substantial diversity of NSP1 observed among group A rotaviruses (GAR) also exists within GBRs and that a high degree of diversity also exists among NSP5 of GBRs, in contrast to GAR NSP5. The NSP1 gene of KB63 contains three ORFs, whereas the NSP1 genes of other GBR strains contain only two. ORFs 2 and 3 of the KB63 gene may be derived from a single ORF corresponding to ORF2 of other GBR strains by the usage of a stop codon created by an upstream single base deletion and single point mutations. In vitro expression studies showed that ORFs 1 and 2, but not 3, of gene 6 can be translated, suggesting that ORF2 may encode a C-terminally truncated, potentially functional product. It may play a role, together with the product of ORF1, in virus replication, as the virus can be passaged further in kids. Similarly, gene 11 can be translated in vitro. Like its counterpart in GARs, the protein encoded by gene 11 was shown to be phosphorylated in vitro. PMID- 10466807 TI - Cellular transcription factors regulate human papillomavirus type 16 gene expression by binding to a subset of the DNA sequences recognized by the viral E2 protein. AB - Human papillomavirus type 16 (HPV-16) is a DNA tumour virus that has been implicated in the development of cervical cancer. The HPV-16 E2 protein binds to four sites that are present upstream of the viral P97 promoter and regulates transcription of the E6 and E7 oncogenes. Here, it is shown that cellular transcription factors bind to two of these E2 sites. One cellular E2 site-binding factor, which is here named CEF-1, binds tightly to E2 site 1. CEF-2, an unrelated cellular E2 site-binding factor, binds tightly to E2 site 3. Transient transfection studies performed in the absence of the E2 protein showed that mutations that blocked the binding of CEF-1 to E2 site 1 or CEF-2 to E2 site 3 significantly reduced P97 promoter activity. Further characterization of CEF-1 indicated that this factor has not previously been identified and that CEF-1 and E2 competed for binding at E2 site 1. PMID- 10466808 TI - Overlapping YY1- and aberrant SP1-binding sites proximal to the early promoter of human papillomavirus type 16. AB - Transcription of oncogenes E6 and E7 of human papillomavirus type 16 (HPV-16) from the P97 promoter is regulated by viral and cellular proteins. The transcription factor YY1 represses transcription through binding to cognate sequences in the long control region (LCR). In HPV-16 DNA from cervical carcinomas, mutations of YY1-binding sites have been identified that increase P97 activity 3-6-fold. A second, SP1-binding site has now been identified in the HPV 16 LCR (nt 7842-7847), which overlaps the YY1-binding site at positions 7840 7848. A point mutation within this YY1 site in viral DNA from a cervical cancer, previously shown to prevent YY1 binding, was shown to increase SP1 binding and P97 activity 4.7-fold. An engineered mutant eliminating SP1 binding showed only 1 to 1.6-fold increased P97 activity. It is concluded that competition between SP1 and YY1 for DNA binding plays a major role in YY1 repression mediated by the binding site at positions 7840-7848. PMID- 10466809 TI - Regulation of cyclin E gene expression by the human papillomavirus type 16 E7 oncoprotein. AB - In this study, we characterized the 5' regulatory region of the murine cyclin E gene and analysed activation of the gene by the E7 oncogene of human papillomavirus type 16 in transfection experiments. We found that the murine cyclin E promoter is composed of multiple regulatory elements, and we present evidence for at least two independent transcription units, designated P1 and P2. Overlapping binding sites for the cellular transcription factors Sp1 and E2F were identified in both promoters, and we found that E2F-mediated activation of transcription is inhibited by Sp1 in cotransfection experiments. The E2F/Sp1 binding sites contribute to transcriptional activation by E7, and the data suggest that the cyclin E gene is rendered E7-inducible through the combination of several cis-acting elements which display only weak intrinsic responsiveness to E7. PMID- 10466810 TI - Improved detection systems for TT virus reveal high prevalence in humans, non human primates and farm animals. AB - TT virus is a newly described agent infecting humans. Initially isolated from a patient (initials T.T.) with unexplained hepatitis, the virus has since been found in both normal and diseased individuals. In the present study, we utilized genomic-length sequences from distinct genotypes of TT virus to design PCR-based assays using conserved oligonucleotide primers from three independent regions of the virus genome. Each of the three assays was found to be superior to the PCR based assays previously published. The most sensitive of the new assays was utilized to demonstrate the prevalence of TT virus to be at least 34.1% in volunteer blood donors, 39.6% in commercial blood donors, 59.6% in non-A-GB hepatitis cases, 81.7% in injectable drug users and 95.9% in haemophiliacs. In an attempt to identify a possible source of human infection, we found TT virus sequences to be present in 19% of chickens, 20% of pigs, 25% of cows and 30% of sheep. Sequence determination and phylogenetic analyses demonstrated that isolates from farm animals were not genetically distinct from those found in humans. This study clearly demonstrates that previously reported PCR assays dramatically underestimate the true prevalence of TT virus within the human population. Due to the high rate of infection in both blood donors and those with non-A-GB hepatitis, these results question the causal role of TT virus in cases of unexplained hepatitis. Further, it is possible that domesticated farm animals serve as a source of human infection. PMID- 10466811 TI - Localization of a unique hepatitis B virus epitope sheds new light on the structure of hepatitis B virus surface antigen. AB - In a search for monoclonal antibodies (MAbs) that can bind hepatitis B virus surface antigen (HBsAg) with amino acid substitutions in the immune dominant 'a' region (escape mutants) we investigated the epitope recognition site of the human MAb 4-7B. Pepscan analysis and experiments with alanine substitution as well as substitutions known from nature pointed to residues 178-186 in the small S protein with the amino acid sequence PFVQWFVGL (key amino acids in bold) as the minimal epitope. Single amino acid substitutions at positions 122(R/K)(d/y), 134(Y/F), 145(G/R), 148(T/A) and 160(K/R)(w/r), representing 'a' region variants in recombinant HBsAg COS-I cells, did not influence binding of MAb 4-7B. Synthetic peptides (residues 175-189) including the 4-7B epitope sequence were able to evoke an anti-HBs response in rabbits. According to established polypeptide models, the 4-7B epitope region is located in the lipid layer of 20 nm HBsAg particles. The present results, however, suggest that residues 178-186 are exposed on the surface of the 20 nm particle. This may change our view of the structure of HBsAg. PMID- 10466812 TI - Kinetics of early molecular events in duck hepatitis B virus replication in primary duck hepatocytes. AB - This paper describes the use of one-step growth conditions to study the kinetics of duck hepatitis B virus (DHBV) replication in primary duck hepatocytes. Synchronized infection was achieved using partially purified DHBV virions at an m.o.i. of 640 DHBV DNA-containing virions per cell, and these conditions were shown to produce a single cycle of infection. In this model, input purified DHBV DNA was rapidly internalized by cells at > or = 0.5 h, and localized to the nucleus by 4 h, but both covalently closed circular (CCC) DNA and single-stranded DNA were not detected until 48 h postinoculation (p.i.), suggesting that there was a > or = 40 h delay between DHBV localization to the nucleus and formation of CCC DNA. In contrast, CCC DNA can be first detected in hepatocytes at 6 h p.i. in in vivo infection of ducks with the same DHBV strain. In an analysis of the nuclear transport of the DHBV genome, release of nuclear viral DNA from a particulate form to a soluble nucleoplasmic form was only 50% complete by 48 h p.i. However, this process occurred simultaneously with genome uncoating since all soluble nucleoplasmic DHBV DNA was free of nucleocapsid material; this suggests that nucleocapsid disassembly and genome uncoating may occur at the nuclear membrane and not within the nucleus. Quantitative analysis demonstrated inefficiency in a number of steps including virus uptake and internalization, translocation of nucleocapsid across the nuclear membrane and antigen expression from intranuclear viral DNA. PMID- 10466813 TI - The vaccinia virus A4OR gene product is a nonstructural, type II membrane glycoprotein that is expressed at the cell surface. AB - Gene A40R from vaccinia virus (VV) strain Western Reserve has been characterized. The open reading frame (ORF) was predicted to encode a 159 amino acid, 18152 Da protein with amino acid similarity to C-type animal lectins and to the VV A34R protein, a component of extracellular enveloped virus (EEV). Northern blotting and S1 nuclease mapping showed that gene A40R is transcribed early during infection from a position 12 nucleotides upstream of the ORF, producing a transcript of approximately 600 nucleotides. Rabbit anti-sera were raised against bacterial fusion proteins containing parts of the A40R protein. These were used to identify an 18 kDa primary translation product and N- and O-glycosylated forms of 28, 35 and 38 kDa. The A40R proteins were detected early during infection, formed higher molecular mass complexes under non-reducing conditions and were present on the cell surface but absent from virions. The proteins partitioned with integral membrane proteins in Triton X-114. Canine pancreatic microsomal membranes protected in vitro-translated A40R from proteinase K digestion, suggesting the A40R protein has type II membrane topology. A mutant virus with the A40R gene disrupted after amino acid 50, so as to remove the entire lectin like domain, and a revertant virus were constructed. Disruption of the A40R gene did not affect virus plaque size, in vitro growth rate and titre, EEV formation, or virus virulence in a murine intranasal model. PMID- 10466814 TI - Herpes simplex virus type 1 infection has two separate modes of spread in three dimensional keratinocyte culture. AB - This study describes the outcome of herpes simplex virus type 1 (HSV-1) infection in an organotypic raft culture of spontaneously immortalized HaCat keratinocytes and human fibroblasts, as related to the virus load and epithelial stratification and differentiation. In this model, a confluent monolayer of HaCat keratinocytes was formed 60 h after seeding. Inoculation of HSV-1 before induction of differentiation by lifting of the culture to the air-liquid interface always resulted in a productive infection, but the virus yield was highest when the inoculation took place 72 h after seeding. Even at 0.1 p.f.u. per culture, the HaCat cultures became HSV positive. Infection of the full-thickness epithelium at 5 p.f.u. per culture resulted in a productive infection of the whole epithelium. The HaCat cells were about 10 times more sensitive to HSV-1 infection than the Vero cells in which the virus stocks were titrated. The raft cultures infected 30 min after lifting were negative by HSV-1 culture, and no HSV-1 antigen was detected by immunocytochemistry. PCR showed the presence of HSV-1 DNA and in situ hybridization showed reactivity with a latency-associated RNA probe, indicating the presence of a non-productive infection. Two different patterns of virus spread in epithelia were found: (i) lateral spread through the superficial layers of the epithelium and (ii) a demarcated infection throughout the whole thickness of the epithelium at the margins of the culture. PMID- 10466815 TI - Nucleolar localization of the UL3 protein of herpes simplex virus type 2. AB - A rabbit polyclonal antiserum was raised against a recombinant 6 x His-UL3 fusion protein expressed in Escherichia coli and used to examine the intracellular localization of the UL3 protein of herpes simplex virus type 2 (HSV-2). The antiserum reacted specifically with 31 and 34 kDa proteins in HSV-2 186-infected Vero cells and with 31 and 35 kDa proteins in UL3-expressing COS-7 cells. The UL3 protein localized both in the cytoplasm and in five to ten bright fluorescent granules in the nucleus close to the nuclear membrane at 4 h post-infection (p.i.). These structures became bigger at 5 h p.i. and showed doughnut-like forms at 6 h p.i. In transfected Vero cells, the UL3 protein localized exclusively in the nucleoplasm and specifically in the nucleolus. Five deletion mutants of the UL3 protein were constructed for transfection assays and the results showed that the region containing amino acids 100-164 was important for nucleolar localization. Moreover, green fluorescent protein (GFP)-targetting experiments showed that the region containing amino acids 100-164 was able to transport non nucleolar GFP to the nucleolus as a fusion protein. PMID- 10466816 TI - Latency-associated transcripts of equine herpesvirus type 4 in trigeminal ganglia of naturally infected horses. AB - Equine herpesvirus type 4 (EHV-4) is a major respiratory pathogen of horses. Unlike most other members of the Alphaherpesvirinae, EHV-4 was regarded as non neurotropic. Here, neural and lymphoid tissues of 17 horses have been analysed post-mortem. EHV-4 DNA was detected in 11 cases (65%) by PCR, exclusively in the trigeminal ganglia. In order to define the transcriptional activity, RNA preparations of 10 EHV-4 DNA-positive ganglia were investigated by nested RT-PCR. EHV-4-specific transcripts derived from genes 63 [herpes simplex virus type 1 (HSV-1) ICPO gene homologue] and 64 (HSV-1 ICP4 gene homologue) were detected in six trigeminal ganglia. In one other case, only gene 64-specific transcripts were present. All of the transcripts proved to be antisense orientated when a strand specific RT-PCR was applied. Type-specific primers for gene 33 (encoding glycoprotein B) served to detect transcripts of an acute EHV-4-infection, which were found in only one of the six ganglia positive for gene 63- and gene 64 specific transcripts. Overall, these studies clearly demonstrate that EHV-4 is latent in trigeminal ganglia. PMID- 10466817 TI - DNA sequence of the UL6 to UL20 genes of infectious laryngotracheitis virus and characterization of the UL10 gene product as a nonglycosylated and nonessential virion protein. AB - The 24 kbp KpnI restriction fragment A from the unique long genome region of infectious laryngotracheitis virus (ILTV, gallid herpesvirus-1) has been sequenced. The analysed region contains 14 open reading frames sharing homology with conserved alphaherpesvirus genes. Arrangement of the UL6 to UL20 homologues of ILTV is almost identical to that found in the herpes simplex virus type 1 genome. As in other herpesviruses the UL15 gene consists of two exons and is expressed from a spliced mRNA. However, the UL16 gene, which is usually localized within the intron sequence of UL15, is not conserved at this position of the ILTV genome. Another unique feature is the absence of any putative N-glycosylation motifs within the deduced ILTV UL10 gene product, which is the homologue of the conserved herpesvirus glycoprotein M. After preparation of a monospecific antiserum, two distinct UL10 proteins with apparent molecular masses of 36 and 31 kDa were identified in ILTV-infected cells as well as in purified virions. None of these UL10 gene products is modified by N- or O-linked glycosylation. Isolation of a green fluorescent protein-expressing UL10 deletion mutant of ILTV revealed that this gene is not required for virus replication in cell culture. PMID- 10466818 TI - Antigenic domain 1 of human cytomegalovirus glycoprotein B induces a multitude of different antibodies which, when combined, results in incomplete virus neutralization. AB - Glycoprotein B (gB, gpUL55) is the major antigen for the induction of neutralizing antibodies against human cytomegalovirus (HCMV), making it an attractive molecule for active and passive immunoprophylaxis. The region between aa 552 and 635 of HCMV gB (termed AD-1) has been identified as the immunodominant target for the humoral immune response following natural infection. AD-1 represents a complex domain which requires a minimal continuous sequence of more than 70 aa for antibody binding. Neutralizing as well as non-neutralizing antibodies can bind to AD-1 in a competitive fashion. The fine specificity of AD 1-binding monoclonal antibodies (MAbs) and affinity-purified human polyclonal antibodies was analysed by using recombinant proteins containing single amino acid substitutions spanning the entire AD-1 domain. Our results revealed that all MAbs had individual patterns of binding to the mutant proteins indicating the presence of a considerable number of distinct antibody-binding sites on AD-1. The neutralization capacity of antibodies could not be predicted from their binding pattern to AD-1 mutant proteins. Polyclonal human antibodies purified from different convalescent sera showed identical binding patterns to the mutant proteins suggesting that the combined antibody specificities present in human sera are comparable between individuals. Neutralization capacities of polyclonal human AD-1 antibodies did not exceed 50% indicating that, during natural infection, a considerable proportion of non-neutralizing antibodies are induced and thus might provide an effective mechanism to evade complete virus neutralization. PMID- 10466819 TI - Epstein-Barr virus lacking latent membrane protein 2 immortalizes B cells with efficiency indistinguishable from that of wild-type virus. AB - Epstein-Barr virus (EBV) is a human herpesvirus that efficiently transforms and immortalizes human primary B lymphocytes. In this study, the role of latent membrane protein 2 (LMP2) in EBV growth transformation was investigated. LMP2 is a virally encoded membrane protein expressed in EBV-immortalized B cells previously shown to be nonessential for EBV transformation. However, a recent study reported that LMP2 may be an important determinant for efficient B cell transformation (Brielmeier et al., Journal of General Virology 77, 2807-2818, 1996). In this study a deletion mutation was introduced into the LMP2 gene using an E. coli mini-EBV construct containing sufficient EBV DNA to result in growth transformation of primary B cells. In an alternative approach, the introduction of the gene encoding the enhanced green fluorescent protein (EGFP) by homologous recombination into the LMP2 gene of EBV strain B95-8, generating the same LMP2 deletion mutation is reported. Careful quantification of B cell transformation using the EGFP+ LMP2- recombinant virus determined that in liquid culture medium or in culture medium containing soft agarose there was no difference in the ability of LMP2- virus to immortalize primary human B cells when compared to that of wild-type virus. PMID- 10466820 TI - Transcriptional activation by the human herpesvirus-8-encoded interferon regulatory factor. AB - Human herpesvirus-8 (HHV-8), a gammaherpesvirus that is thought to be the viral aetiologic agent of Kaposi's sarcoma and primary effusion lymphoma, encodes a homologue to cellular interferon regulatory factors (IRFs). The HHV-8 IRF homologue (vIRF; ORF K9) has previously been shown to inhibit gene induction by interferons and IRF-1 and to transform NIH3T3 cells or Rat-1 cells. Additionally, expression of antisense to vIRF in BCBL-1 cells results in the repression of certain HHV-8 genes, suggesting that vIRF may also positively regulate gene expression. We demonstrate that vIRF activates transcription when directed to DNA by the GAL4 DNA-binding domain. GAL-vIRF truncation constructs that individually are incapable of activating transcription can cooperate in transactivation when coexpressed in HeLa cells, suggesting that multiple regions of vIRF are involved in transactivation. These studies broaden the potential mechanisms of action of vIRF to include transcriptional activation as well as transcriptional repression. PMID- 10466821 TI - A naturally occurring deleted form of RNA 2 of Potato mop-top virus. AB - A spontaneous deletion in RNA 2 of Potato mop-top virus (PMTV) was identified by RT-PCR. The deletion occurred reproducibly during manual passage of two isolates of PMTV and during fungal inoculation of plants with viruliferous soil. The borders of the deletion were conserved in all instances and sequence analyses showed that a contiguous segment of 2113 nucleotides was deleted internally from the genomic RNA 2, leaving the 5'- and 3'-terminal sequences. RT-PCR experiments also showed that the deletion was present in preparations of PMTV particles. PMID- 10466822 TI - A short open reading frame terminating in front of a stable hairpin is the conserved feature in pregenomic RNA leaders of plant pararetroviruses. AB - In plant pararetroviruses, pregenomic RNA (pgRNA) directs synthesis of circular double-stranded viral DNA and serves as a polycistronic mRNA. By computer-aided analysis, the 14 plant pararetroviruses sequenced so far were compared with respect to structural organization of their pgRNA 5'-leader. The results revealed that the pgRNA of all these viruses carries a long leader sequence containing several short ORFs and having the potential to form a large stem-loop structure; both features are known to be inhibitory for downstream translation. Formation of the structure brings the first long ORF into the close spatial vicinity of a 5' proximal short ORF that terminates 5 to 10 nt upstream of the stable structural element. The first long ORF on the pgRNA is translated by a ribosome shunt mechanism discovered in cauliflower mosaic (CaMV) and rice tungro bacilliform viruses, representing the two major groups of plant pararetroviruses. Both the short ORF and the structure have been implicated in the shunt process for CaMV pgRNA translation. The conservation of these elements among all plant pararetroviruses suggests conservation of the ribosome shunt mechanism. For some of the less well-studied viruses, the localization of the conserved elements also allowed predictions of the pgRNA promoter region and the translation start site of the first long ORF. PMID- 10466823 TI - Sequence changes in six variants of rice tungro bacilliform virus and their phylogenetic relationships. AB - The DNA of three biological variants, G1, Ic and G2, which originated from the same greenhouse isolate of rice tungro bacilliform virus (RTBV) at the International Rice Research Institute (IRRI), was cloned and sequenced. Comparison of the sequences revealed small differences in genome sizes. The variants were between 95 and 99% identical at the nucleotide and amino acid levels. Alignment of the three genome sequences with those of three published RTBV sequences (Phi-1, Phi-2 and Phi-3) revealed numerous nucleotide substitutions and some insertions and deletions. The published RTBV sequences originated from the same greenhouse isolate at IRRI 20, 11 and 9 years ago. All open reading frames (ORFs) and known functional domains were conserved across the six variants. The cysteine-rich region of ORF3 showed the greatest variation. When the six DNA sequences from IRRI were compared with that of an isolate from Malaysia (Serdang), similar changes were observed in the cysteine-rich region in addition to other nucleotide substitutions and deletions across the genome. The aligned nucleotide sequences of the IRRI variants and Serdang were used to analyse phylogenetic relationships by the bootstrapped parsimony, distance and maximum-likelihood methods. The isolates clustered in three groups: Serdang alone; Ic and G1; and Phi-1, Phi-2, Phi-3 and G2. The distribution of phylogenetically informative residues in the IRRI sequences shared with the Serdang sequence and the differing tree topologies for segments of the genome suggested that recombination, as well as substitutions and insertions or deletions, has played a role in the evolution of RTBV variants. The significance and implications of these evolutionary forces are discussed in comparison with badnaviruses and caulimoviruses. PMID- 10466825 TI - Role of the 3' untranslated region of baculovirus p10 mRNA in high-level expression of foreign genes. AB - The p10 gene of Autographa californica nucleopolyhedrovirus has two putative AATAAA polyadenylation signals. The downstream signal is used predominantly, as was determined by analysing 3' cDNA ends. This downstream motif is followed by a GT-rich sequence, known to be important for efficient polyadenylation in mammalian systems. To analyse the importance of polyadenylation for p10 gene expression, recombinant viruses with altered 3' untranslated regions (UTRs) were tested using chloramphenicol acetyltransferase (CAT) as a reporter. Surprisingly, after inactivation of the downstream AATAAA motif, CAT expression remained at the same high level as observed with a wild-type 3' UTR. Polyadenylation occurred 24 28 nucleotides further downstream, probably due to an ATTAAA sequence motif. Replacing the p 10 3' UTR with the SV40 early terminator sequence as part of an hsp70-lacZ-SV40 gene cassette, which is commonly used in baculovirus expression vectors, resulted in a reduction in reporter gene expression. Polyadenylation occurred far more efficiently in the original p10 3' UTR than in the SV40 terminator sequence, as was shown by testing the SV40 terminator separately. These results indicate that in order to obtain high levels of foreign gene expression, vectors that provide a wild-type p10 3' UTR are to be preferred over those containing the hsp70-lacZ-SV40 gene cassette. Comparison of the p10 genes of various baculoviruses showed the presence of at least one AATAAA or ATTAAA motif in combination with a GT-rich sequence in the 3' UTR, suggesting an evolutionary conservation of these two elements, thereby maintaining the high level of p10 gene expression. PMID- 10466824 TI - Rapid generation of genetic heterogeneity in progenies from individual cDNA clones of peach latent mosaic viroid in its natural host. AB - Viroids, small single-stranded circular RNAs endowed with autonomous replication, are unique systems to conduct evolutionary studies of complete RNA genomes. The primary structure of 36 progeny variants of peach latent mosaic viroid (PLMVd), evolved from inoculations of the peach indicator GF-305 with four individual PLMVd cDNAs differing in their pathogenicity, has been determined. Most progeny variants had unique sequences, revealing that the extremely heterogeneous character of PLMVd natural isolates most probably results from the intrinsic ability of this RNA to accumulate changes, rather than from repeated inoculations of the same individual trees under field conditions. The structure of the populations derived from single PLMVd sequences differed according to the observed phenotype. Variant gds6 induced a reproducible symptomatic infection and gave rise to a more uniform progeny that preserves some parental features, whereas variant gds15, which induced a variable phenotype, showed a more complex behaviour, generating two distinct progenies in symptomatic and asymptomatic individual plants. Progenies derived from variants esc10 and Is11, which incited latent infections, followed a similar evolutionary pattern, leading to a population structure consisting of two main groups of variants, one of which was formed by variants closely related to the parental sequence. The evolution rate exhibited by PLMVd, considerably higher than that reported for potato spindle tuber viroid, may contribute to the fluctuating symptomatology of the severe PLMVd natural isolates. However, the polymorphism observed in PLMVd progenies does preserve some structural and functional elements previously proposed for this viroid, supporting the fact that they act as constraints limiting the genetic divergence of PLMVd quasispecies generated de novo. PMID- 10466826 TI - Identification and functional analysis of a putative non-hr origin of DNA replication from the Spodoptera littoralis type B multinucleocapsid nucleopolyhedrovirus. AB - A putative non-hr origin of DNA replication was identified in the Spodoptera littoralis multinucleocapsid nucleopolyhedrovirus (SpliNPV) genome by transient replication assays. The putative SpliNPV ori was mapped to the PstI-J fragment between 75.1-77.9 map units in the SpliNPV genome. While the DNA sequence of the putative SpliNPV ori aligned with regions within the non-hr oris of Autographa californica, Orgyia pseudotsugata and Spodoptera exigua multinucleocapsid nucleopolyhedroviruses, it has limited DNA sequence identity with these elements. The sequence of the putative SpliNPV non-hr ori fragment contains a unique distribution of imperfect palindromes, multiple direct repeats and putative transcription factor-binding sites. Transient expression assays indicated that the putative SpliNPV ori fragment repressed SpliNPV lef-3 promoter-mediated luciferase reporter gene expression. However, the putative SpliNPV ori fragment itself was capable of directing luciferase expression in the absence of a recognizable baculovirus promoter element in an orientation-independent fashion, suggesting that DNA sequence motifs within its sequence can activate transcription. Gel mobility shift analyses confirmed that proteins within nuclear extracts from both uninfected and virus-infected cells bound with specificity to the putative SpliNPV ori fragment. PMID- 10466827 TI - PrP (prion) gene expression in sheep may be modulated by alternative polyadenylation of its messenger RNA. AB - Scrapie-associated fibrils and their major protein component, PrP or prion protein, accumulate in the brains and some other tissues of all species affected by transmissible spongiform encephalopathies or prion diseases. To investigate the role of PrP gene expression in the hosts of these diseases, we have analysed some characteristics of PrP gene RNA transcripts in sheep and cattle tissues and made comparisons with PrP RNA transcripts in human and mouse tissues. Two PrP messenger RNAs of 4.6 kb and 2.1 kb, the result of alternative polyadenylation, were found first in sheep peripheral tissues and also occurred at low levels in sheep brain and bovine tissues, but not in human and mouse tissues. Our results from transfection assays of murine neuroblastoma cells with constructs expressing different regions of ovine PrP messenger RNA revealed the presence of sequences in the 3' untranslated region of the gene that modulate protein synthesis. PMID- 10466828 TI - Manufacturers asked to improve packaging for OP sheep dips. PMID- 10466829 TI - Survey of infectious agents involved in acute respiratory disease in finishing pigs. AB - Outbreaks of respiratory disease constitute a major health problem in herds of finishing pigs and their aetiology often remains unclear. In this study, 16 outbreaks of respiratory disease with acute clinical signs in finishing pigs were investigated to determine which infectious agents were involved. From each herd four diseased and two clinically healthy pigs were examined pathologically and for the presence of viruses, bacteria and mycoplasmas. In addition, paired blood samples from 10 groupmates of the diseased pigs were tested for antibodies against commonly known causal agents of respiratory disease. A clear diagnosis was possible in 12 of the 16 outbreaks. Seven were due to an infection with influenza virus and five were due to an infection with Actinobacillus pleuropneumoniae. A combination of influenza virus and A pleuropneumoniae may have caused one other outbreak, but no clear cause could be established for the other three outbreaks. PMID- 10466830 TI - Hypertrophic osteodystrophy in six weimaraner puppies associated with systemic signs. AB - Six weimaraner puppies, five of which were genetically related, showed systemic signs associated with hypertrophic osteodystrophy, including fever and involvement of the gastrointestinal, respiratory or nervous systems, in addition to the metaphyseal lesions. In five of the dogs the clinical signs developed less than 10 days after they had been vaccinated with a modified live virus vaccine. Radiographic findings suggested that both the hindlimbs and forelimbs were equally involved in the disease process. Abnormal haematological findings included leucocytosis with neutrophilia and monocytosis, and there was a consistent increase in the activity of alkaline phosphatase. Serum protein electrophoretic studies of three of the dogs revealed hypogammaglobulinaemia and abetaglobulinaemia in two of them. Conservative treatment with rest and non steroidal anti-inflammatory drugs had little effect, and treatment with corticosteroids appeared to give the best results. PMID- 10466831 TI - Clinicopathological features of equine primary hepatic disease: a review of 50 cases. AB - The clinicopathological features of 50 cases of equine hepatic disease were reviewed. There was a wide range of clinical signs and at least 50 per cent of the animals exhibited either dull demeanour, anorexia, abdominal pain, cerebral dysfunction and/or weight loss. Life-threatening complications of hepatic failure recorded were: gastric impaction in 10 cases, bilateral laryngeal paralysis in seven cases and coagulopathy in five cases. All the cases had high activities of gamma-glutamyl transferase (GGT) and most had high activities of glutamate dehydrogenase (GLDH) and high concentrations of bile acids. Fewer of the horses had abnormal concentrations of bilirubin, albumin and globulin. The horses that were euthanased or died had significantly higher concentrations of GGT, GLDH and bile acids than the survivors. There were biochemical data for 18 cases with signs of hepatic encephalopathy, all of them had plasma ammonia levels greater than 90 micromol/litre but this was not significantly correlated with the clinical severity of the condition. Half of the cases with hepatic encephalopathy were hyperglycaemic, none was hypoglycaemic, and none had abnormally low levels of plasma urea. PMID- 10466832 TI - Identification and incidence of porcine circovirus in routine field cases in Quebec as determined by PCR. PMID- 10466833 TI - Vestibular disease in two horses: a case of mycotic otitis media and a case of temporohyoid osteoarthropathy. PMID- 10466834 TI - Tuberculosis in cattle and badgers. PMID- 10466835 TI - Tuberculosis in cattle and badgers. PMID- 10466836 TI - Pet travel scheme. PMID- 10466837 TI - Cydectin 1 per cent injectable solution for sheep. PMID- 10466838 TI - Urine analysis in equine grass sickness. PMID- 10466839 TI - False acacia poisoning in horses. PMID- 10466840 TI - Ragwort poisoning in horses. PMID- 10466841 TI - Society of Veterinary Assistants? PMID- 10466842 TI - Testing 40 predictions from the transtheoretical model. AB - Sensitive measurement of behavior change requires dependent measures that are sensitive to the whole spectrum of change, not just a single aspect of change. Traditional outcome variables such as point prevalence for smoking cessation focus on a single discrete event and ignore all other progress. Alternatively, the criterion measurement model (CMM) is an approach that posits a three construct outcome model (habit strength, positive evaluation strength, and negative evaluation strength), where different constructs are sensitive to change for different aspects of the temporal domain. In this article, a series of 40 differential a priori predictions were tested using a large representative sample of smokers. The focus was on the prediction of specific effect sizes rather than statistical significance. A series of comparisons involving stage transitions was examined using five variables representing the three CMM constructs. The predictions involved movement from one of three initial stages (precontemplation, contemplation, and preparation) to stage membership 12 months later. Thirty-six of the 40 predictions were confirmed, indicating that the outcome model has strong construct validity and accurately reflects movement between the stages of change. PMID- 10466843 TI - Treatment career snapshots: profiles of first treatment and previous treatment clients. AB - To examine the substance abuse recovery process from a treatment-career perspective, this study compared the profiles of clients entering treatment for the first time versus those returning to treatment. Clients (N = 7,092) presenting for publicly funded treatment were assessed as part of the St. Louis Target Cities project. Previous treatment clients were more likely to have more severe substance abuse problems, additional psychiatric problems, and greater problems in other life areas. Comorbid psychiatric problems and motivation to change were identified as areas of focus for clinical intervention. Discriminant analysis yielded a significant function that accounted for 6.5% of the variance between groups. Results suggest that change occurs via multiple processes. PMID- 10466844 TI - Psychiatric severity and behavior change in alcoholism: the relation of the transtheoretical model variables to psychiatric distress in dually diagnosed patients. AB - Treatment programming for individuals diagnosed with a chronic mental illness and an alcohol use disorder could be enhanced by employing techniques that focus on those change process variables that are most strongly related to psychiatric distress. Prochaska and DiClemente's transtheoretical model (TTM) provides a useful framework within which to study these relations. The associations between psychiatric severity and the TTM constructs of stages and processes of change, decisional balance, temptation, and self-efficacy were measured among 132 alcohol dependent patients in a public mental health clinic's outpatient dual diagnosis program. Participants' scores on the Temptation subscale of the Alcohol Abstinence Self-Efficacy Questionnaire are strongly related to psychiatric severity: The more psychiatric distress a person is experiencing, the more he or she is tempted to drink, particularly in situations that trigger negative affect. Decisional balance considerations are also related to psychiatric severity: The higher participants scored on the Global Severity Index of the Brief Symptom Inventory, the more importance they placed on the negative aspects, or cons, of drinking. Subjects with more psychiatric distress also scored higher on the maintenance stage of change subscale, possibly indicating an increased fear of relapse and struggle to maintain sobriety. PMID- 10466845 TI - Use of the BULIT bulimia screening questionnaire to assess risk and progress in weight management for overweight women who weight cycle. BULImia Test. AB - A bulimia screening questionnaire was examined for usefulness in assessing risk for bulimia and measuring progress in reducing binge eating in overweight women who weight cycle. In two studies the BULIT test was used to screen for risk for bulimia. Study 1 was a descriptive study of motivations for overeating in normal weight women compared to overweight women who weight cycled. In Study 2, overweight women who weight cycled were examined at baseline, 6, and 12 months for effectiveness of a clinical treatment strategy to reduce binge eating. In Study 1, BULIT scores were statistically significantly higher for overweight compared to normal weight subjects. In Study 2, subjects' BULIT scores were lower after using a long term clinical treatment strategy to gain control of eating. The BULIT test was useful for specifying four categories of eating patterns to assess risk for bulimia and progress in reducing binge eating. PMID- 10466846 TI - Alcohol administration methodology 1994-1995: what researchers do and do not report about subjects and dosing procedures. AB - This article reviews which parts of the alcohol administration methodology were reported in 90 alcohol studies on humans published from 1994 to 1995. although several subject characteristics such as gender were regularly reported, other variables that can influence pharmacokinetics and responses to alcohol were not consistently reported. It is suggested that guidelines for reporting human alcohol administration studies be created and that journal editors and funding agencies require submissions to meet minimum standards for describing the study methodology. Potential guidelines are presented. PMID- 10466847 TI - Psychosocial correlates of nicotine dependence among men and women in the U.S. naval services. AB - This study assessed the prevalence and psychosocial correlates of clinical criteria-based diagnoses of nicotine dependence in men and women in the U.S. naval services. Analyses were based on data from the 1995 Perceptions of Wellness and Readiness Assessment, a population-based self-report survey of 9,856 active duty Navy and Marine Corps members worldwide, specifically focusing on a subsample of those who completed the telephone version of the Quick Diagnostic Interview Schedule (n = 782). Twenty-five percent of all telephone interview volunteers met diagnostic criteria for lifetime nicotine dependence, and 15% met criteria for nicotine dependence in the last year. There was no difference in nicotine dependence by job classification (traditional vs. nontraditional) among women. Women who were abused prior to entering the military were almost three times as likely to be nicotine dependent as those not abused. Men who had been exposed to combat or violence were twice as likely to be nicotine dependent as those not exposed. PMID- 10466848 TI - Psychological distress and adaptational problems associated with discontinuation of benzodiazepines. AB - This study compared subjects who had received standard tapered withdrawal of benzodiazepine (BZD) (group 1) with a group with comparable diagnosis still receiving BZD (group 2) and a control group of comparable diagnosis not yet receiving treatment (group 3). Sixty subjects aged 21-65 years with a diagnosis of nonpsychotic anxiety or insomnia were included. The assessment of psychological distress and quality of life was timed to coincide with the maximum immediate effect of BZD discontinuation, as calculated according to drug half life. Subjects diagnosed with insomnia reported lower distress in all three groups. The pattern of distress experienced by group 1 was closer to group 3 than to group 2, indicating the potential importance of re-emergence of anxiety. High neuroticism, lower education level, and lower quality of life were associated with higher levels of distress during withdrawal. PMID- 10466849 TI - Alcohol cue reactivity and private self-consciousness among male alcoholics. AB - Recent alcohol cue exposure studies have noted that not all alcoholics demonstrate increased reactivity when presented with alcohol-related cues. This study examined the relationship of private self-consciousness (PSC) to subjective, self-report measures of reactivity and measures of negative mood states that involve a focus on internal processes. These subjective measures of reactivity were contrasted with salivary reactivity, an objective measure of reactivity which does not require individual self-report or awareness. A cue reactivity assessment was administered to 47 men meeting DSM-III-R criteria for a diagnosis of alcohol dependence. Our hypothesis, that PSC would predict urge reactivity status and greater levels of negative mood states, was supported. Urge reactors were more likely to be high in PSC, while the proportions of salivary reactors and nonreactors were not significantly different between the high and low PSC groups. Although regression analyses indicated that PSC did not significantly predict urge to drink alcohol, it did predict angry/frustrated mood and sad/depressed mood at the first alcohol trial. These results suggest that individuals high in PSC may benefit more from cue exposure-based treatment, as they are more likely to be urge reactors and to evidence negative mood reactivity. Low PSC individuals may be at higher risk for relapse given they are less able to recognize internal reactions signaling the presence of a high-risk alcohol use situation, and therefore less likely to mobilize coping responses. PMID- 10466850 TI - Characteristics of smokeless tobacco users seeking treatment. AB - Previous studies have described smokeless tobacco (ST) treatment seekers with minimal detail. In the present study, ST users (N = 402), who enrolled in a ST cessation treatment study, were asked to complete an extensive questionnaire that inquired about their ST use patterns, use of other tobacco products, extent of dependence, previous attempts to quit, situations associated with use and support for quitting. The results showed that this population experiences a high level of nicotine exposure and physical dependence on ST. ST use frequently is associated with negative affect situations and times of hunger. A high frequency of users have smoked cigarettes as well as cigars. A supportive social environment for ST cessation exists for these individuals. These results have implications for ST treatment content. PMID- 10466851 TI - Dose-effect relationship between overdose mortality and prescribed methadone dosage in low-threshold maintenance programs. AB - The present study evaluates the effectiveness of low-threshold methadone maintenance in reducing overdose mortality. In a prospective cohort study conducted in Amsterdam, 498 Dutch injecting drug users (IDU) provided 1,969 person years of follow-up (1989-1995). Forty-four IDU died in this period, 15 due to illicit drug overdose. Compared to IDU not in maintenance, the adjusted relative risk for overdose mortality among those receiving 5-50 mg, 55-70 mg, and 75+ mg were 0.35, 0.13, and 0.11, respectively (p < .05). Also current injection use and HIV-seropositivity were independent predictors for overdose mortality. Whereas previous studies indicated effectiveness of methadone-assisted detoxification and high-dose maintenance programs in reducing mortality, the present study findings suggest that low-threshold maintenance programs also reduce overdose mortality, with higher dosages being most protective. PMID- 10466852 TI - Overlapping addictions and self-esteem among college men and women. AB - To examine whether there is a tendency for individuals to be multiply addicted, overlapping addictions to common substances (alcohol, caffeine, chocolate, cigarettes) and activities (exercise, gambling, Internet use, television, video games) were studied in 129 college men and women. Contrary to previous research, moderate to large correlations were found, both within and between substances and activities. Self-esteem was positively related to exercise but unrelated to the remaining addictions. Several gender differences in addictive tendencies were also revealed: Men scored higher than women on addiction to alcohol, cigarettes, gambling, television, and Internet use, but women scored higher on caffeine and chocolate. The results have implications for theories of addiction and suggest new directions for the study of addiction among normally functioning young adults. PMID- 10466854 TI - A comparison of three "interventions" on pretreatment dropout rates in an outpatient substance abuse clinic. AB - Studies examining pretreatment dropout in outpatient substance abuse clinics have consistently demonstrated that fewer days between phone contact and scheduled intake appointment result in higher client show rates. To compare time to intake with two other interventions hypothesized to increase show rates, individuals (N = 128) seeking treatment were randomly assigned to one of four groups. Individuals in Group 1 were scheduled within 48 hours of the telephone intake. Individuals in Groups 2, 3, and 4 were scheduled 48 hours or more after the telephone intake and received either a reminder call 24 hours prior to their appointment (Group 2), an appointment card and clinic brochure in the mail (Group 3), or no intervention (Group 4). The results show that Group 1 had a higher show rate as compared to the other three groups, and receiving a reminder call or an appointment card did not improve show rates beyond that of the no-contact control group. Implications for intake procedures are discussed. PMID- 10466853 TI - Smoking and diabetes: readiness for change and provider advice. AB - Smoking is a serious health risk, particularly for people with diabetes. This study was designed to examine important aspects of smoking in a large group of individuals with diabetes. A survey was mailed to 2,056 individuals with diabetes. The variables examined were the stages of change for smoking, prevalence of quitting advice given by health care providers, and the patterns of readiness for change. The majority (57.8%) of current smokers were in the precontemplation stage. Comparisons on the stage of change indicated that more individuals with Type 2 diabetes have quit while there are more current smokers among those with Type 1 diabetes. Comparisons on current smokers indicated no differences on stage of change across the Type 1 and Type 2 groups, across three subgroups of individuals with Type 2 diabetes, or across duration of diabetes. Those who reported that they were given cessation advice were further along in the stages of change. These results suggest that the majority of individuals with diabetes who smoke are in the precontemplation stage of change and provider advice is important in moving smokers toward change. The current findings underscore the importance of assessing stage of change and providing stage matched interventions when working with smokers with diabetes. PMID- 10466855 TI - Risk factors for suicide attempts among low-income women with a history of alcohol problems. AB - This study used bivariate and multivariate analyses to examine nine risk factors for suicide attempts among 80 women (51 attempters, 29 controls) with a history of alcohol problems who were recruited from a large, inner-city hospital. Prior studies established that each of the examined factors increased the risk for suicidal behavioral, but these studies have varied according to whether or not they consisted exclusively of persons with alcohol problems. Whereas eight of the nine factors were bivariately associated with suicide attempt status in this study, only three factors (hopelessness, recent interpersonal loss, childhood trauma) remained significant in the multivariate analysis. These findings illuminate the importance of using multivariate analyses when aiming to identify factors that uniquely increase the risk for suicidal behavior among persons with alcohol problems. PMID- 10466856 TI - One-year outcome of an integrative inpatient treatment for dual diagnosis patients. AB - The purpose of this study was to evaluate a 4-month dual diagnosis inpatient treatment program that was based on integrative models for patients with substance use and psychiatric disorders. At intake and at 1-year follow-up, dual diagnosis patients (N = 52) were assessed on housing and subsistence level, substance use and psychiatric symptoms. At 1-year follow-up, dual diagnosis patients reported a higher level of housing and subsistence, and less intensive psychiatric symptoms. No change occurred on frequencies of substance use. General improvement may occur even if psychotropic substances are used. Controlled studies are needed on extended inpatient treatment for dual diagnosis patients and outcomes in longer follow-ups. PMID- 10466857 TI - Experimental investigation of I-123 iodoamphetamine in the detection of lung cancer. AB - Experimentally we investigated the detection of lung cancer with N-isopropyl-p-I 123-iodoamphetamine (I-123 IMP). Various tumors including Lewis lung cancer were used as tumor models. Serial images were obtained. Biodistribution of Lewis lung cancer was performed. In Lewis lung cancer good visualization as in B-16 melanoma and high tumor accumulation were found with IMP. In conclusion, due to its greater accumulation almost equivalent to that in melanotic melanoma, I-123 IMP may have a role in the detection of lung cancer. PMID- 10466858 TI - Molecular genetic characteristics of lung cancer--useful as real' tumor markers? AB - The increased knowledge about the molecular mechanisms leading to the development of a tumor has opened new horizons for basic and applied research. Lung cancer is among the tumor entities with the highest incidence and mortality rates. Recently new drugs and therapeutic options for patients with lung cancer were developed. These developments demand new and improved techniques for the sensitive and specific detection of lung tumor cells. Some of the molecular genetic features of lung tumor cells are summarized and the possibilities to use these characteristics as new tumor markers are discussed. PMID- 10466859 TI - Prognostic value of the serum tumour markers Cyfra 21-1 and tissue polypeptide antigen in malignant mesothelioma. AB - In malignant mesothelioma, survival is claimed to be related to age, duration of symptoms, performance status, histological subtype, stage and platelet count. However the exact prognostic value of these factors is still a matter of debate. We studied the two cytokeratin markers, Cyfra 21-1 and Tissue polypeptide antigen (TPA) for their significance in predicting survival retrospectively in 52 patients. Cyfra 21-1 and TPA were elevated in 26 (50%) and 30 (58%) patients, respectively, and were highly correlated (r = 0.98). Univariate analysis of data from 51 patients, showed a relation with survival for performance status (P = 0.010), thoracic pain (P = 0.014), platelet count (P = 0.027), Cyfra 21-1 (P = 0.002) and TPA (P = 0.003). Multivariate analysis identified independent prognostic significance for performance status, platelet count and Cyfra 21-1. In addition to performance status ( < 80 vs. > 80) the cytokeratin markers identified patients with good prognosis in a log rank test. Values of Cyfra 21-1 and TPA are significantly correlated. PMID- 10466860 TI - Does an incomplete interlobar fissure influence survival or recurrence in resected non-small-cell lung cancer? AB - OBJECTIVE: There have been various reports on prognostic factors in non-small cell lung cancer (NSCLC) under a complete resection, but an incomplete interlobar fissure has not been discussed yet. We attempted to clarify whether this influences survival or recurrence. PATIENTS AND METHODS: From 1981 to 1994, 239 patients (43%) who had a single lobectomy with mediastinal lymph node dissection were pathologically diagnosed as stage IA/IB or IIA/IIB (excluding T3N0M0). These patients were divided in two groups: patients with a complete interlobar fissure group (group L); and patients with an incomplete one (group NL). Postoperative survivals and cancer-free periods were retrospectively assessed. Patients' characteristics had no statistical difference between groups L and NL by N category. RESULTS: The 5-year survival was 77.0% in group L-N0, 79.3% in group NL N0, 48.7% in group L-N1, and 66.2% in group NL-N1. No statistical difference was found between groups L-N0 and NL-N0, L-N1 and NL-N1, L-T1N0 and NL-T1N0, L-T1N1 and NL-T1N1, L-T2N0 and NL-T2N0, and L-T2N1 and NL-T2N1. The 5-year-relapse-free survival was 81.2% in group L-N0, 85.4% in group NL-N0, 69.4% in group L-N1, and 72.2% in group NL-N1. No statistical difference was found between groups L-N0 and NL-N0, L-N1 and NL-N1, L-T1N0 and NL-T1N0, L-T1N1 and NL-T1N1, L-T2N0 and NL T2N0, and L-T2N1 and NL-T2N1. There was no statistical difference in recurrent sites between groups L-N0 and NL-N0, and L-N1 and NL-N1. CONCLUSION: We conclude that the incomplete interlobar fissure does not influence the prognosis of resected stage I or II NSCLC (excluding T3N0M0). PMID- 10466861 TI - Phase I/II dose finding study of paclitaxel and carboplatin in advanced non-small cell lung cancer. AB - INTRODUCTION: This phase I study was designed to establish the maximum tolerated dose (MTD) of the carboplatin paclitaxel combination, given without routine growth factor support to previously untreated patients with stage IIIB and IV non small cell lung cancer. PATIENTS AND METHODS: Fifty patients (one stage IIIa, 31 stage IIIb and 18 stage IV) were sequentially assigned to one of 14 treatment groups in which paclitaxel and carboplatin were administered in doses ranging from 130 to 235 mg/m2 and from 230 to 375 mg/m2 , respectively. Paclitaxel was administered as a 3-h intravenous infusion using non-polyvinylchloride tubing and connectors. The carboplatin infusion, over 30 min, was administered at the completion of the paclitaxel infusion. RESULTS: The MTD for the combination has been reached at 235 mg/m2 of paclitaxel and at 375 mg/m2 of carboplatin. The combination shows a good safety profile with very few and brief episodes of neutropenia without any infectious episodes. At the doses tested thrombocytopenia did not occur at all. Among 47 assessable patients there was one complete response and 17 partial responses for an overall response rate of 38%. There has been a tendency to a dose-response relationship for the combination with only six partial responses (27%) reported in 22 patients who received paclitaxel at doses < or = 195 mg/m2 and carboplatin at doses < 350 mg/m2 and 12 partial responses in 25 patients (48%) receiving paclitaxel > 195 mg/m2 and carboplatin > or = 350 mg/m2, respectively. The median event-free survival time is 33 weeks (range, 4 156 +). With a minimum follow up duration of 57 weeks the median overall survival time is 51.81 weeks (range, 7-162 +) and the 1-year survival rate is 49%. CONCLUSION: In advanced NSCLC the carboplatin-paclitaxel combination can be safely administered at the doses of 375 and 225 mg/m2 every 4 weeks, it appears to be active and well tolerated. PMID- 10466862 TI - A genomic approach to scoliosis pathogenesis. AB - Genetic predisposition contributes to scoliosis in humans. Two syndromes of primary scoliosis occur--congenital scoliosis, which presents at birth, often associated with other abnormalities, and idiopathic scoliosis which becomes apparent between infancy and adolescence. Little is known regarding the genetic transmission of scoliosis risk. Data gleaned from mouse mutations provide a valuable supplement to human family studies. More than 50 mouse mutations include scoliosis, kyphosis, or tail kinks as a phenotype; the locations of the human homologues for 28 of these can be predicted on the basis of synteny conservation. Some mouse mutations are either more penetrant or more fully expressed in one sex. The mouse data provide a basis both for optimism and for caution in understanding human scoliosis. Mouse models provide insight into mechanisms underlying spinal curvature and help direct searches for genes important in human disease. Four types of defects account for most mouse scoliosis: defects of cell cell communication, intracellular signal transduction, matrix protein synthesis, and matrix protein metabolism. Mouse data suggest that at least two types of heterogeneity complicate genetic analysis: locus heterogeneity, in which lesions of distinct genes lead to a similar phenotype, and allelic heterogeneity, in which the phenotypes arising from alleles of a single gene differ. By focusing initial studies on multiplex families with apparent simple Mendelian inheritance the effect of heterogeneity is minimized. PMID- 10466863 TI - Factors associated with staff stress and work satisfaction in services for people with intellectual disability. AB - Staff stress and morale have been identified as major issues affecting the quality of services for people with intellectual disability. The present study investigates factors directly and indirectly associated with staff general distress, job strain and work satisfaction amongst staff in services for people with intellectual disability. As part of a large-scale survey of staff in services for people with intellectual disability, information was collected from 450 staff concerning general distress, job strain and work satisfaction, and a wide range of factors potentially associated with these outcomes. Path analyses revealed that three factors accounted for 28% of the variance in general distress scores: (I) wishful thinking, (2) stress linked to work-home conflict and (3) role ambiguity. Six factors accounted for 50% of the variance in job strain scores: (I) wishful thinking, (2) stress linked to a lack of staff support, (3) alienative commitment, (4) role ambiguity, (5) stressors linked to a low status job and (6) working longer contracted hours. Six factors accounted for 66% of the variance in work satisfaction scores: (I) stress linked to a low status job, (2) support from supervisors, (3) influence over work decisions, (4) alienative commitment, (5) support from colleagues and (6) older staff age. A range of factors indirectly associated with the three outcome measures was also identified. The models of general distress, job strain and work satisfaction empirically derived in the present study confirm and extend previous research in this area. The implications for organizations and future research are discussed. PMID- 10466865 TI - Is megalencephaly specific to autism? AB - Several recent reports have described the presence of increased head circumference (megalencephaly) in patients with autism. Although some studies have described reports of megalencephaly in other disorders such as schizophrenia in adults, few such studies have been performed in children and adolescents. In the present study, the authors compared 20 subjects with autism/ pervasive developmental disorder (DSM-IV; all males; mean age = 10.9 years) with 20 controls with attention deficit hyperactivity disorder (DSM-IV; all males; mean age = 11.1 years). Four subjects and five controls had evidence of megalencephaly. In addition to their core symptoms, the autistic subjects with megalencephaly were hyperactive and impulsive. These findings suggest that megalencephaly may not be specific to autism, and when present, it may index the presence of additional symptoms such as hyperactivity and impulsivity. PMID- 10466864 TI - Stress and residential staff who work with people who have an intellectual disability: a factor analytic study. AB - The present study describes the development of a questionnaire, and the subsequent collection of data, to gather information on the demands and supports which influence stress (as measured by anxiety and depression scales) in direct care staff who work with people with intellectual disability. The results from the questionnaire were also used to explore the relationships between the factors derived from the questionnaire and to consider how these may be influential in building organizational models. The questionnaire was specifically constructed to measure the demands and supports experienced by direct care staff. The participants included 216 staff working in community residential services for people with intellectual disability. These individuals included staff working for two British health trusts and similar staff working for one local authority. The questionnaire consisted of 33 demands items and 23 supports items. Separate factor analyses were conducted on the demands and supports scales. Analyses of the reliability and validity of the resulting factors were conducted. Stepwise multiple regressions were conducted to further explore the relationship of factors with the measures of anxiety and depression. Four factors were extracted from the demands scale and three factors from the support scale. There was some concern over levels of reliability and validity for individual factors; however, the reliability of the main scales appeared to be satisfactory. All but one support factor correlated significantly with levels of anxiety and depression. The factors explained a modest amount of the variance in the regressions. While there are some concerns about the psychometric properties of the questionnaire, it is argued that the scales and factors can be used to confirm and further understanding of the relationship between groups of demands, supports and stress in staff. The usefulness of the questionnaire as a means of diagnosing specific sources of demand and support is considered. It is suggested that the questionnaire could form the basis for assessment and subsequent intervention in houses where staff are reporting relatively high levels of anxiety and depression. PMID- 10466866 TI - Cervical spondylarthrotic myelopathy with early onset in Down's syndrome: five cases and a review of the literature. AB - Progressive walking difficulties and bladder dysfunction may be attributed to Alzheimer disease or atlanto-axial subluxation in people with Down's syndrome (DS). The present authors describe five patients with DS suffering from the above symptoms as a result of cervical spondylarthrotic myelopathy. Clinical and radiological data were collected from all patients with DS who underwent surgery for cervical spondylarthrotic myelopathy at the Leiden University Medical Centre during the period between 1991 and 1995. Five patients with DS (four males and one female) were identified. Their mean age at diagnosis was 42 years. The main clinical features were weakness of the arms and legs, ataxic gait, hyperreflexia and bilateral Babinski signs. Radiological examination showed spondylarthrosis, compression of the spinal cord and myelomalacia. The mean delay in diagnosis was 3 years. All five individuals showed clinical stabilization after laminectomy. Cervical spondylarthrotic myelopathy seems a rather frequent disorder in DS, occurring at a relatively young age. Early diagnosis may prevent irreversible neurological deficits. PMID- 10466867 TI - The Thomas S. case: report on progress with court compliance issues. AB - The present paper is a brief summary of the progress which has been made in the Thomas S. case, a U.S. Federal Court lawsuit The suit focused on adults with intellectual disability who had been inappropriately placed in state psychiatric hospitals in conditions which violated their constitutional rights. These constitutional rights include safety, protection from harm, treatment under safe conditions, freedom from undue restraint, and minimally adequate habilitation or treatment The present report summarizes the progress made by the 239 class members after they had received a Thomas S. service plan for 6-24 months. Overall, the results suggest that the class members have made significant progress in several areas. Two areas needing continued attention are the challenging behaviours of some class members and their lack of employment opportunities. PMID- 10466868 TI - Visuo-spatial and linguistic abilities in a twin with Williams syndrome. AB - The present study reports a case of dizygotic twins, one boy with Williams syndrome (WS) and one typically developing girl, and compares their neuropsychological profiles. The goal of the present authors was to verify whether the child with WS displayed a cognitive profile which is unique to the syndrome. Several tests designed to assess visuo-perceptual, visuo-motor, linguistic and memory abilities were administered to both children when they were 10.9 years old. Compared to his sister, the boy with WS displayed a homogeneous developmental delay in both non-verbal and verbal abilities. He achieved a level of performance similar to his sister only in facial recognition, phonological word fluency and memory for phonologically similar words. Furthermore, despite the overall delayed performance of the boy, both the twins displayed a cognitive profile characterized by strength in lexical comprehension and relative weakness in visuo-motor abilities. PMID- 10466869 TI - Excess of non-right handedness among individuals with intellectual disability: experimental evidence and possible explanations. AB - Seventy-three subjects with mild and moderate intellectual disability (ID) of an unknown/idiopathic nature, who had no record of specific deficits in motor control, and 73 normal controls, matched for chronological age and sex, underwent evaluation of handedness. The results confirmed the previously reported excess of non-right handedness among subjects with ID, and suggested that this finding might be indicative of a deviant cerebral organization or atypical lateralization of cerebral representation of function in this group. However, it was noted that further research is needed to clarify how far this is a function of level of motor skill and how far is a syndrome-specific pathological phenomenon. PMID- 10466870 TI - Fragile-X syndrome, Down's syndrome and autism: awareness and knowledge amongst special educators. AB - Fragile-X syndrome is the commonest cause of inherited intellectual disability. There is good evidence for a behavioural phenotype. This has implications for school staff using standard educational techniques. Similarly, autism is known to create particular educational requirements. The present study examined the awareness and knowledge of fragile-X syndrome, Down's syndrome and autism amongst staff in special and mainstream education. One hundred and two special school staff and 40 mainstream school staff completed questionnaires. Most staff offered a variety of features 'typical' of Down's syndrome and autism. In contrast, staff knew less about fragile-X syndrome. Specific knowledge about the learning styles of these children was very poor, but was associated with having taught an affected child. Mainstream and special school staff offered similar levels of knowledge for all three conditions. Staff did not demonstrate a sufficiently specialized knowledge of fragile-X syndrome to ensure that the special educational needs of these children were being met fully. PMID- 10466871 TI - Mediator analysis: an overview of recent research on carers supporting people with intellectual disability and challenging behavior. AB - Families and front-line care staff are often key agents in helping deliver behavioural interventions to people with intellectual disability and challenging behaviour. The present paper reviews factors which may impact on the effectiveness of such interventions. Both PsychLit and MedLine were used to conduct an automated literature search which included the terms 'learning disability', 'mental handicap', 'mental retardation', 'intellectual disability', 'challenging behaviour', 'families' and 'staff. This search was substantially augmented by additional citations obtained from books and journal articles. The impact of attitudes, beliefs and emotional states as setting conditions for interventions was highlighted. A variety of more general setting conditions related to intervention effectiveness were also identified. Implications for both clinical practice and future research were discussed. PMID- 10466872 TI - Liver transplantation: lessons from the Nordic experience. PMID- 10466873 TI - Helicobacter pylori test-and-treat' strategy for dyspeptic patients. PMID- 10466874 TI - Bile reflux into the esophagus. Bilitec 2000 measurements in normal subjects and in patients after Nissen fundoplication. AB - BACKGROUND: Bilitec 2000 is a new spectrophotometric system that can detect bilirubin within the esophagus. The aim of this study was to test this new system in vitro and in vivo with special attention to patients operated on laparoscopically for gastroesophageal reflux with Nissen fundoplication. METHODS: In vitro studies of gallbladder bile dilution curves at different pHs were performed with a combined pH and Bilitec probe and by investigation of the drift of the Bilitec system. In vivo studies were performed in 34 normal individuals and 12 patients operated on with a laparoscopic Nissen fundoplication. Intra/intervariations were evaluated in normal individuals. RESULTS: The Bilitec system was able to detect changes in absorbance value within the range 0.14-0.6. Absorbance values were about 30% lower in an acid environment. The percentage of the total time with bile reflux was a median (range) of 3.9 (0-49.6) in normal individuals, with an intravariation within the intervariation, and 8.7 (0-36.6) in patients after a Nissen fundoplication. These values did not differ significantly. Eighty-two per cent of the normal subjects had some degree of bile reflux, if an absorbance value > or = 0.14 was accepted as the threshold value for the presence of bile reflux. CONCLUSIONS: Bile reflux in normal individuals is frequent and not different from that in patients after a Nissen fundoplication. Surprisingly, the latter had bile reflux but no or minimal acid reflux. Factors other than bilirubin may interfere with the measurements. PMID- 10466875 TI - Helicobacter pylori infection, pattern of gastritis, and symptoms in erosive and nonerosive gastroesophageal reflux disease. AB - BACKGROUND: The aim of this study was to evaluate the prevalence of Helicobacter pylori infection and the characteristics of gastritis and symptoms of patients with erosive and nonerosive gastroesophageal reflux disease (GERD). METHODS: We studied 202 consecutive patients with a diagnosis of GERD (symptoms score and endoscopy): group A (n = 110), erosive GERD; group B (n = 92), nonerosive GERD; 200 patients with upper abdominal complaints without abnormalities at endoscopy (functional dyspepsia, group C); and 200 asymptomatic controls tested for H. pylori serum antibody (group D). Antral and body biopsy specimens were taken for histology and the rapid urease test in groups A, B, and C. RESULTS: The prevalence of H. pylori infection was higher in groups B and C (62% and 55%, respectively) than in A and D (36% and 40%) (P < 0.05). In positive patients H. pylori colonization and gastritis grade scores in the gastric body were higher in nonerosive than in erosive GERD and functional dyspepsia (P < 0.05). No differences in H. pylori colonization or gastritis grades were found in the antrum. Fifty-nine patients with nonerosive GERD (64%) and 42 with erosive GERD (38%) showed other dyspeptic symptoms associated with reflux symptoms (P < 0.05). CONCLUSIONS: H. pylori prevalence is higher in patients with nonerosive GERD than in normal subjects and in patients with erosive GERD and similar to that of patients with dyspepsia. Patients with nonerosive GERD often show dyspeptic symptoms and higher H. pylori colonization and inflammation grades in the proximal stomach. Our data support the hypothesis that in GERD H. pylori gastritis may, on the one hand, protect against the development of esophageal erosions and, on the other, contribute to the esophageal hypersensitivity to acid which is a feature of GERD. PMID- 10466877 TI - The effects of omeprazole on intragastric pH, intestinal motility, and gastric emptying rate. AB - BACKGROUND: The present study was designed to investigate whether omeprazole changes the characteristics and thereby the functions ascribed to fasting intestinal motility, postprandial motility, postprandial pH, and gastric emptying. METHODS: Ten healthy subjects were investigated. The studies were performed after 10 days of treatment with 40 mg omeprazole daily/placebo. Duodenal pressures and intragastric pH were detected by strain-gauge transducers and a pH electrode attached to a miniature computer. The meal consisted of an omelette labelled with 99mTc-sulphur colloids followed by 150 ml water labelled with 111In-diethylenetriamine pentaacetic acid. RESULTS: The difference in fasting intragastric pH between the two series was highly significant. The profile from the placebo series showed a relationship between phase activity and pH. The pH increased from phase I (median, 1.3; 95% confidence interval (CI), 0.9 1.6) to a maximum at 25% (1.8 (0.9-2.1)) and 50% (1.6 (1.1-3.8)) of cycle duration and decreased thereafter until the end of the cycle. The profile from the omeprazole series showed significantly higher values during the entire cycle but no relationship between phase activity and pH. Pretreatment with omeprazole was followed by a delay in gastric emptying of liquid at 30 min (64% (49%-66%) (omeprazole series) versus 78% (67%-83%); P < 0.01) and solid at 180 min (71% (48%-86%) (omeprazole series) versus 96% (87%-100%); P < 0.01). There was no significant difference in duration of postprandial motility (305 min (157-350 min) (omeprazole) versus 259 min (129-403 min)). CONCLUSIONS: Omeprazole eliminates the temporal relationship between intragastric pH and characteristics of the migrating motor complex and induces a delay in gastric emptying of both liquid and solid. A non-significant increase in duration of postprandial motility may represent a type-II error. PMID- 10466876 TI - Effect of an aluminum hydroxide-magnesium hydroxide combination drug on adhesion, IL-8 inducibility, and expression of HSP60 by Helicobacter pylori. AB - BACKGROUND: Co-magaldrox (Maalox) is used world-wide as an antacid and as a cytoprotective agent for gastritis and peptic ulcer diseases. We examined the effects of co-magaldrox on Helicobacter pylori. METHODS: Adhesion of H. pylori to human gastric epithelial cells (MKN45) was evaluated by flow cytometry. Morphologic changes in H. pylori caused by co-magaldrox were determined by scanning electron microscopy. Induction of interleukin-8 (IL-8) from MKN45 cells was examined with enzyme-linked immunosorbent assay, and the intracellular and extracellular expression of heat-shock protein 60 (HSP60) was analyzed with sodium dodecyl sulphate-polyacrylamide gel electrophoresis and flow cytometry. RESULTS: Adhesion of H. pylori to MKN 45 cells was significantly inhibited by 1.25%-5% comagaldrox. H. pylori aggregated with co-magaldrox according to an electron microscopic examination. IL-8 secretion from MKN45 cells after H. pylori infection was also inhibited by co-magaldrox. Extracellular expression of HSP60 on the surface of H. pylori was decreased after treatment with comagaldrox, whereas the intracellular synthesis of HSP60 was not. HSP60-induced IL-8 secretion was significantly inhibited by co-magaldrox in a dose-dependent manner. CONCLUSIONS: These results show that co-magaldrox suppressed the expression of the following virulence factors: adhesion, IL-8 inducibility, and expression of extracellular HSP60. Therefore, co-magaldrox is a potent anti-H. pylori and cytoprotective drug. PMID- 10466878 TI - Five-year audit of ambulatory 24-hour esophageal pH-manometry in clinical practice. AB - BACKGROUND: Esophageal function testing was developed to aid diagnosis in patients with negative endoscopy. Although combined 24-h esophageal pH-manometry is now commercially available, its routine clinical effectiveness has not yet been studied. METHODS: From 1992 to 1996 we evaluated 303 consecutive patients who were first-time referrals to our unit for 24-h esophageal pH-manometry. The referral indications were gastroesophageal reflux disease, 47.2%; dysphagia, 18.5%; non-cardiac chest pain, 14.9%; connective tissue disease, 13.2%; and symptomatic patients after antireflux surgery, 6.3%. RESULTS: Overall, esophageal function testing altered the diagnosis of 44% of the patients, confirmed it in 38%, and specifically changed the management of 66%. The final clinical 'diagnosis' was reflux disease, 54% (32% with non-specific esophageal motility disorder); connective tissue disease, 9.9%; achalasia, 9.6%; other specific esophageal motility disorders, 3.3%; non-specific esophageal motility disorders, 6.9%; and normal, 16.2%. The cost per testing was estimated to be US$305 and per change in management US$465. CONCLUSION: Combined 24-h pH-manometry has been shown to be a useful and cost-effective test for the management of selected patients in whom the primary investigation was insufficient. PMID- 10466879 TI - Abnormal response to cholinergic stimulation in the circular muscle layer of the human colon in diverticular disease. AB - BACKGROUND: Diverticular disease is characterized by the occurrence of small herniations of the colonic mucosa, through the external muscle coats of the colon. The muscle wall is thickened, high intraluminal pressures can be recorded, and often constipation develops. The aim of the present study was to investigate whether an abnormality in the electric myogenic control activity could be found to help explain the etiology and symptoms of the disease. METHODS: Electric activity was studied by extracellular electrodes on tissues from both the circular and the longitudinal muscle of the colon from 12 patients. RESULTS: In tissues from 10 patients a distinctly abnormal response to cholinergic stimulation was observed. A characteristic development of bursts of action potentials did not develop; instead, slow-wave activity of relatively low frequency was maintained throughout the period of stimulation. This slow-wave activity showed a lack of synchronization. CONCLUSIONS: The results indicate that, in diverticular disease, local changes in electric activity occur that change the response to cholinergic stimulation. When this happens, development of periodic bursts of action potentials normally associated with propulsive activity do not develop, favoring segmental contractile activity associated with low frequency slow-wave activity. PMID- 10466880 TI - Regional distribution and localization of zinc and metallothionein in the intestine of rats fed diets differing in zinc content. AB - BACKGROUND: Zinc (Zn) is protective and enhances epithelial repair in gut diseases. In this study we investigate the localization and distribution of Zn and its binding protein, metallothionein (MT), in the gut of rats fed diets varying in Zn content. METHODS: Male-Sprague Dawley rats were fed low, normal, high, or excess Zn in their diets (10, 100, 400, or 1000 mg Zn/kg, respectively) and killed 7 days later. Blood, liver, and gut tissues were collected. Tissue Zn was determined with atomic absorption spectrophotometery and MT with a Cd/haem affinity assay. Zn and MT were immunohistochemically localized in the small intestinal wall with zinquin and an anti-MT antibody. RESULTS: Most Zn in the intestinal wall was present in the mucosal scrapings, with 94% membrane-bound and 6% cytosolic, irrespective of dietary Zn. MT levels increased in all gut regions at dietary Zn levels above 100 mg Zn/kg. MT was 40% higher in the ileum than in other gut regions in rats fed low- and normal-Zn diets. The Zn content of the ileum was also 20% higher than that of other gut regions in rats fed low-, normal , or high-Zn diets. Zn and MT were colocalized in the base of the intestinal crypts, most visibly in the ileum. CONCLUSION: Mucosal cytosolic Zn and MT concentrations are increased only at high or excessive Zn intakes in all gut regions except the ileum, which can respond to a lower Zn intake. As the cytosolic Zn pool most likely influences mucosal protection and repair mechanisms, it is proposed that an increased MT may indicate the adequacy of oral Zn therapy in gut disease. PMID- 10466881 TI - Bone loss in patients with inflammatory bowel disease is less than expected: a follow-up study. AB - BACKGROUND: In spite of the accumulating evidence of an increased prevalence of osteopenia and osteoporosis in patients with inflammatory bowel diseases (IBD), the time course of bone loss is not well described, and there is little knowledge about factors indicating an increased risk of rapid bone loss. METHODS: We conducted a follow-up study in 80 IBD patients (45 men and 25 premenopausal and 10 postmenopausal women), 19 with ulcerative colitis and 61 with Crohn disease, with a mean follow-up time of 568 +/- 60 days, to assess bone loss, risk factors of rapid bone loss, and value of bone markers to predict bone loss. Bone mineral density was measured by dual-energy X-ray absorptiometry, bone formation by bone alkaline phosphatase (BAP), and bone resorption by N-terminal telopeptide of type I collagen (NTX) and free deoxypyridinoline (DPD). RESULTS: Bone density changes per year were 0.46% +/- 3% at the spine, 0.06% +/- 5.1% at the femoral neck, 1.1% +/- 7.7% at the triangle of Ward, and -0.52% +/- 1.86% at total body level. Type and duration of disease, sex, age, and level of NTX, DPD, and BAP at base line did not show significant differences between patients who lost and those who did not lose bone mass. Bone loss was significantly higher in patients with (n = 28) than in those without steroids (n = 52) at the femoral neck and Ward triangle but not at the spine and total body. CONCLUSIONS: Change in bone mass in IBD patients during short-term follow-up is low on average, but there is great heterogeneity within the population, which cannot be explained by the use of steroids alone. Bone loss cannot be predicted by analysis of bone markers. PMID- 10466882 TI - Polymorphism of heat-shock protein gene HSP70-2 in Crohn disease: possible genetic marker for two forms of Crohn disease. AB - BACKGROUND: Previous reports have shown that heat-shock protein 70 (HSP70) may be associated with Crohn disease. However, genetic analysis of the HSP70 gene in Crohn disease has not been done. The aim of this study is to investigate whether HSP70-2 gene polymorphism is involved in susceptibility to Crohn disease in the Japanese population and whether it correlates with clinical manifestation of the disease. METHODS: A total of 108 genetically unrelated patients with Crohn disease and 108 healthy controls were typed for HSP70-2 PstI polymorphism by restriction fragment length polymorphism analysis (alleles A and B). Patients with Crohn disease were classified into two types: either perforating or non perforating. RESULTS: Genotype and allele frequency did not differ between patients and controls. In patients with Crohn disease, allele A frequency was significantly higher in the non-perforating than in the perforating type (P = 0.02). When patients with Crohn disease of more than 6.7 years' duration were assessed, the differences in genotype and allele frequency between the two groups became more significant (P < 0.001 and P < 0.001, respectively). CONCLUSIONS: These data suggest that HSP70-2 gene polymorphic allele A is a possible genetic marker of less severe clinical phenotype in Japanese patients with Crohn disease. PMID- 10466883 TI - Stapled functional end-to-end anastomosis versus sutured end-to-end anastomosis after ileocolonic resection in Crohn disease. AB - BACKGROUND: The aim of this retrospective study was to compare complications and anastomotic recurrence rates after stapled functional end-to-end versus conventional sutured end-to-end anastomosis after ileocolonic resection in Crohn disease. METHODS: Between 1988 and 1997, 123 patients underwent ileocolonic resection for Crohn disease. Forty-five patients underwent stapled functional end to-end anastomosis (stapled group), and 78 underwent sutured end-to-end anastomosis (sutured group). RESULTS: The stapled anastomosis has been more frequently used during the past 3 years; between 1995 and 1997 it was used in 33 (83%) of 40 patients, compared with only 12 (14%) of 83 patients between 1988 and 1994. There was one anastomotic leak (2%) in the stapled group, compared with six (8%) in the sutured group. The overall complication rate was significantly lower in the stapled group (7% versus 23%, P = 0.04). In the stapled group only one patient required reoperation for ileocolonic anastomotic recurrence, compared with 26 in the sutured group. The cumulative 1-, 2- and 5-year rates for ileocolonic recurrences requiring surgery in the stapled group were 0%, 0%, and 3%, which were significantly lower than the 5%, 11%, and 24% in the sutured group (P = 0.007 by log-rank test). CONCLUSIONS: Although the follow-up duration was short in the stapled group, these results suggest that stapled functional end-to end ileocolonic anastomosis is associated with a lower incidence of complications and that early anastomotic recurrence is less common than after sutured end-to end anastomosis. However, a randomized trial would be necessary to draw clear conclusions. PMID- 10466884 TI - Liver transplantation in the Nordic countries, 1982-1998: changes of indications and improving results. AB - BACKGROUND: Liver transplantation has become an established therapeutic option for patients with life-threatening liver disease. The aim of the present study was to analyse the results of and developments in liver transplantation in the Nordic countries during a 15-year period. METHODS: Data on all patients receiving a liver allograft in the Nordic countries during 1982-98 and waiting list data for all patients listed for a liver transplantation after 1989 were obtained from the Nordic Liver Transplantation Registry. RESULTS: A total of 1485 first liver transplantations were performed during 1982-98. The annual number of first liver transplantations increased steadily up to 1993, thereafter remaining around 150 170 per year. There are major differences between countries both in the number of transplants adjusted to populations performed per year, with more than twice as many performed in Sweden as in Norway, and in the relative distribution of patients in accordance with diagnosis. The number of patients more than 60 years old increased and comprised 13%-14% of the total patient population during 1996 98. Primary biliary cirrhosis, primary sclerosing cholangitis, acute hepatic failure, malignant liver disease, and alcoholic cirrhosis are the five most frequent diagnoses. The over-all 1-year patient survival probability has increased from 66% among patients receiving a transplant in 1982-89 to 83% in 1995-1998. The waiting time remains stable, with a median waiting time of 35 days during 1990-98. The mortality of patients while on the waiting list is 7.4% and is not increasing. CONCLUSION: Results of liver transplantation in the Nordic countries are very similar to those obtained in other countries. Waiting time and mortality remain low. There are, however, major differences between the countries both as to the number of transplantations performed and as to distribution of diagnoses. PMID- 10466885 TI - Prediction of hepatic encephalopathy in paracetamol overdose: a prospective and validated study. AB - BACKGROUND: Paracetamol overdose may cause hepatic encephalopathy (HE). This condition demands specialized care and, in some instances, liver transplantation evaluation. No model is available for predicting HE. We aimed to set up and validate a model for predicting the occurrence of HE in paracetamol overdose. METHODS: Prospectively, 161 patients with single-dose paracetamol overdose and no HE (defined as hepatic coma grade II or more) on admission were studied during a 26-month period. Patients admitted during the first 13-month period constituted a learning set to construct a model to predict the occurrence of HE. Patients admitted in the second 13-month period constituted the validation set. Serial biochemical variables (measured twice daily), the time line after the overdose, and demographic data were used for univariate testing, and significant factors were assessed in various multiple logistic regression analyses. RESULTS: Thirty two patients (20%), 15 in the first period and 17 in the second, developed HE grade II. The best model (the highest chi-square) for HE included: log10 (hours from overdose to antidote treatment), log10 (plasma coagulation factors on admission), and platelet count hours from overdose (chi-square = 41.2, P < 0.00001). In the validation set 88% (confidence interval (CI), 64%-99%) of the patients who developed HE were correctly predicted by the constructed model, whereas 90% (CI, 79%-96%) of the patients in the non-HE group were correctly predicted. CONCLUSIONS: The constructed model for predicting HE in paracetamol overdose proved sensitive and accurate in the validation set and should be valuable for transferring high-risk patients to a liver intensive care unit/transplantation facility. PMID- 10466886 TI - Exocrine pancreatic function and fat malabsorption in human immunodeficiency virus-infected patients. AB - BACKGROUND: Nutrients malabsorption frequently occurs in human immunodeficiency virus (HIV)-infected patients, but very few studies have investigated exocrine pancreatic digestive capacity in these patients. We therefore evaluated the frequency of exocrine pancreatic impairment and its eventual relation with fat malabsorption in HIV-infected patients. METHODS: Thirty-five HIV-infected patients (30 male, 5 female: mean age +/- standard deviation, 33.6 +/- 7.2 years) and 51 sex- and age-matched controls without gastroenterologic diseases were studied. In all subjects fecal elastase 1 (EL-1) was assayed, and fecal fat excretion was evaluated with the steatocrit test. RESULTS: Nineteen of 35 (54%) HIV-infected patients showed subnormal EL-1 values, whereas all the controls had normal values; furthermore, EL-1 values were significantly lower in patients than in controls: mean (95% confidence intervals), 207 ( 164-251 ) microg/g versus 312 (291-332) microg/g (P < 0.0001). Increased fecal fat excretion was observed in almost all (25 of 35) HIV-infected patients, and an inverse but not significant correlation was found between fecal EL-1 and steatocrit values. No association was found between reduced fecal EL-1 and the severity of HIV disease or nutritional and immunologic status. Opportunistic infections and drug administration had no influence on EL-1 concentrations in stools. CONCLUSIONS: Reduced exopancreatic function is frequent in HIV-infected patients but does not seem to be a major factor contributing to fat malabsorption. PMID- 10466887 TI - Ageing and reoxygenation injury in the liver. PMID- 10466888 TI - Age-related cochlear hair cell loss is enhanced in mice lacking copper/zinc superoxide dismutase. AB - Age-related hearing loss in humans and many strains of mice is associated with a base-to-apex gradient of cochlear hair cell loss. To determine if copper/zinc superoxide dismutase (Cu/Zn SOD) deficiency influences age-related cochlear pathology, we compared hair cell losses in cochleas obtained from 2-, 7-, and 17- to 19-month-old wild type (WT) mice with normal levels of Cu/Zn SOD and mutant knockout (KO) mice with a targeted deletion of Sod1, the gene that codes for Cu/Zn SOD. WT and KO mice exhibited similar patterns of hair cell loss with age, i.e., a baso-apical progression of hair cell loss, with greater loss of outer hair cells than inner hair cells. Within each age group, the magnitude of loss was much greater in KO mice compared to WT mice. The results indicate that Cu/Zn SOD deficiency potentiates cochlear hair cell degeneration, presumably through metabolic pathways involving the superoxide radical. PMID- 10466889 TI - Age-related psychomotor and spatial learning deficits in 129/SvJ mice. AB - The 129 mouse strain has been widely used to construct mutations that model behavioral aging in humans. The current study found significant age-related declines in both psychomotor and swim maze performance of 5-, 17-, and 27-month old 129/SvJ mice. However, the age differences in swim maze acquisition were inconsistent with poor performance in the probe trial which assesses spatial memory. This inconsistency may result from the high degree of genetic polymorphisms and age-related visual pathology which afflicts this mouse strain. Therefore, we concluded that 129/SvJ mice present a problematic model of mammalian cognitive aging and involve a risk for behavioral contamination in studies involving mutant mice derived from this strain. PMID- 10466890 TI - Age and species-dependent differences in the neurokinin B system in rat and human brain. AB - Neurokinin B and its cognate neurokinin-3 receptor are expressed more in the forebrain than in brain stem structures but little is known about the primary function of this peptide system in the central processing of information. In general, few studies have specifically addressed age-related changes of tachykinins, notably the changes in number and/or distribution of the neurokinin B-expressing and neurokinin-3 receptor-bearing neurons. Data on functions and changes of neurokinins in physiological aging are limited and apply mainly to the substance P/neurokinin-1 receptor system. In the present study, we analyzed neurokinin B/neurokinin-3 receptor system in young (5 months) versus middle aged (15 months) and old rats (23-25 months) and also in aging human brains. For the majority of the immunohistochemically examined regions of the rat brain, there was no statistically significant change in neuronal number and size of the neurokinin B and neurokinin-3 receptor staining. In the adult human brain, there was no age-associated change of the number or size of neurokinin-B-positive neurons. However, we found a major decline in number of neurokinin-3 receptor expressing neurons between young/middle aged (30 years to 69 years) versus old (70 years and older) adults. Interestingly, numbers of neurokinin-3 receptor positive microglia increased whereas the neurokinin-3 receptor-positive astrocytes remained unchanged in both aging rat and human brains. Finally, in addition to assessing the morphological and quantitative changes of the neurokinin B/neurokinin-3 receptor system in the rat and human brain, we discuss functional implications of the observed interspecies differences. PMID- 10466891 TI - Subunit and region-specific decreases in nicotinic acetylcholine receptor mRNA in the aged rat brain. AB - We have investigated possible changes in the mRNA levels for several alpha and beta subunits of the nicotinic acetylcholine receptor (nAChR) and the level of binding for nicotinic ligands in 7- to 32-month-old rats. Alpha4 and beta2, and to a lesser extent alpha6 and beta3, mRNA levels showed decreases between 20 and 30% at 29 months of age which in some areas reached 50% at 32 months of age. Alpha7 showed a small increase from 7 to 14 months and then a progressive decrease from 14 to 32 months down to the 7-month levels. 3H-epibatidine binding did not significantly change from 7 to 32 months of age in rat tel- and diencephalon. Binding in the substantia nigra was exceptional in that it showed a significant decrease starting from 23 months of age. 125I-alpha-bungarotoxin binding showed a pattern of change which roughly paralleled that of alpha7 mRNA. These findings show that an alteration in some steps of nAChR biosynthesis takes place during aging, which may be related to functional changes in nicotinic transmission. PMID- 10466892 TI - Primate-like amyloid-beta sequence but no cerebral amyloidosis in aged tree shrews. AB - A central pathological feature of Alzheimer's disease is the profuse deposition of amyloid-beta protein (Abeta) in the brain parenchyma and vessel walls. Abeta also forms deposits in the brains of a variety of mammals, including all aged non human primates studied to date. The sequence of Abeta in these animals is identical to that in humans. No Abeta deposits have been found in the brains of wild-type rats and mice, suggesting that the three amino acid differences between their Abeta and that of amyloid-bearing mammals impedes the fibrillogenicity of Abeta. Analysis of the primary sequence of the beta-amyloid precursor protein in tree shrews revealed a 98% similarity and 97% identity with the human protein. Furthermore, the predicted amino acid sequence of Abeta in tree shrews is identical to that in humans. However, immunohistochemical analysis failed to reveal beta-amyloid deposits in the neural parenchyma or vasculature of eight aged (7-8 years) tree shrews (Tupaia belangeri). The lack of correlation between the Abeta sequence and amyloid formation suggests that other factors contribute to cerebral amyloid deposition in aged animals. PMID- 10466893 TI - Cerebral amyloid angiopathy in an aged great spotted woodpecker (Picoides major). AB - A male great spotted woodpecker (Picoides major), which was at least 16 years old, died due to general weakening. Cerebral vascular walls, including capillaries, were positively stained with Congo red with green-gold birefringence, and some of which showed a severe deposition of the Congophilic materials resulting in a corona-like fibrillar radiating structure. The Congophilic materials were positive for beta amyloid protein, but negative for prion protein. Only a few senile plaque-like structures were observed in the cortex by PAM stain and beta amyloid immunostain. The present case is the first observation of cerebral amyloid angiopathy in avian species and will indicate the presence of such age-related cerebral lesions also in birds. PMID- 10466894 TI - On the survival time of a tangled neuron in the hippocampal CA4 region in parkinsonism dementia complex of Guam. AB - In diseases such as the Parkinson dementia complex of Guam (PDC) or Alzheimer's disease, susceptible neurons develop intracellular tangles (iNFTs) and then die, leaving behind extracellular tangles (eNFTs). We performed counts of healthy neurons, iNFTs, and eNFTs in the hippocampus of Guamanian Chamorros who were neurologically normal or who suffered from PDC. The total of surviving and dead neurons in the CA4 region was remarkably constant from case to case, indicating that eNFTs are not phagocytosed. Since cases of recent PDC showed only marginal tangle formation in CA4, we concluded that tangle development in CA4 commenced close to the onset of the disease. Based on this assumption, as well as the further assumption that the average rate of tangle development and the average lifetime of a tangled neuron do not alter as the disease progresses, we derived equations to determine the average lifetime of tangled neurons. The results varied from 0.13 years for the most rapidly progressing case to 7.98 years for the most slowly developing case. The average for 8 cases was 2.51 years. PMID- 10466895 TI - Hypothesis: amyloid beta-peptides truncated at the N-terminus contribute to the pathogenesis of Alzheimer's disease. AB - Fragments of amyloid beta-peptide truncated at the N-terminus are present in AD brain early in the course of Alzheimer's disease. It is proposed that these species contribute to the pathogenesis of AD, possibly through destabilization of elements of the neuronal cytoskeleton by small fibrillar deposits. PMID- 10466896 TI - Truncating the amyloid cascade hypothesis: the role of C-terminal Abeta peptides in Alzheimer's disease. PMID- 10466897 TI - N-terminus truncated beta-amyloid peptides and C-terminus truncated secreted forms of amyloid precursor protein: distinct roles in the pathogenesis of Alzheimer's disease. PMID- 10466898 TI - Familial Alzheimer disease: changes in Abeta production may indicate a disturbance in protein transport or function caused by pleiotropic effects of FAD mutations. PMID- 10466899 TI - The shorter amyloid cascade hypothesis. PMID- 10466900 TI - DNA damage and DNA repair in the nervous system. PMID- 10466901 TI - Effect of over-the-counter cimetidine on phenytoin concentrations in patients with seizures. AB - OBJECTIVE: To determine the effects of the maximum recommended over-the-counter (OTC) cimetidine dosage on phenytoin concentrations in ambulatory seizure patients on long-term phenytoin therapy. METHODS: Adults with seizure disorders requiring phenytoin therapy were recruited. Trough total phenytoin concentrations were measured initially and once weekly for six weeks. All assays were performed using Biotrack patient-side cartridges. After a two-week baseline period, patients took cimetidine 200 mg twice daily for two weeks. Toxicity was monitored via weekly neurologic examinations and midweek telephone surveys. Patients were asked to return to clinic weekly during a two-week cimetidine washout period. RESULTS: Nine patients entered and completed the study. All but two patients took other anticonvulsants known to interact with phenytoin (carbamazepine, n = 5; phenobarbital, n = 2). No adverse effects or changes in seizure frequency were reported. Paired Student's t-tests revealed no significant difference between serum phenytoin concentrations before (12.3+/-3.2 mg/L [mean +/- SD]) and after (12.8+/-4.0 mg/L) two weeks on the OTC cimetidine regimen. No differences were noted in estimated pharmacokinetic parameters (maximum metabolic rate, Michaelis Menten constant) for the same time periods (paired Student's t-test, p > 0.05). The Biotrack assay had an r2 = 0.7311 (p < 0.001, two-sided) when compared with TDx. CONCLUSIONS: It is possible that the lack of change in phenytoin concentrations was a result of the low daily dosage of cimetidine used or other factors related to the "real world" setting of the study. However, the potential for a serious drug interaction occurring in patients taking long-term oral phenytoin and OTC cimetidine appears to be small. PMID- 10466902 TI - Accuracy, clinical correlation, and patient acceptance of two handheld prothrombin time monitoring devices in the ambulatory setting. AB - OBJECTIVE: To evaluate the accuracy, clinical correlation, ease of use, and patient acceptance of the Coaguchek and the ProTime Microcoagulation System as compared with standard laboratory methods for prothrombin time determination. METHODS: A total of 30 prothrombin times, expressed as international normalized ratios (INRs), were determined by each handheld device for comparison with standard laboratory testing. Accuracy was evaluated by calculating the absolute difference for each pair of INR values. Clinical correlation was defined as an INR obtained by the handheld monitor that would have resulted in the same therapeutic decision as the INR obtained by the standard laboratory method. Subjects were surveyed to determine which method of INR determination they preferred and their reasons for that preference. RESULTS: Accuracy was superior with the Coaguchek monitor. The absolute difference (mean +/- SD) in the laboratory and Coaguchek INRs was 0.28+/-0.23 (p = 0.96). The absolute difference (mean +/- SD) in the laboratory and the ProTime Microcoagulation System INRs was 0.56+/-0.34 (p < 0.001). For clinical correlation, two out of 24 (8.3%) INRs with the Coaguchek were sufficiently different from the laboratory INR to have resulted in a different therapeutic decision, compared with 12 out of 24 (50%) with the ProTime Microcoagulation System (p < 0.005). Of subjects surveyed, 77.8% preferred the finger stick method. CONCLUSIONS: The Coaguchek was superior to the ProTime Microcoagulation System in accuracy, clinical correlation, and ease of use. The study also showed that patients preferred capillary blood sampling by finger puncture over venipuncture for INR monitoring. PMID- 10466903 TI - Absorption of rectally administered phenytoin: a pilot study. AB - OBJECTIVE: To test the hypothesis that rectally administered phenytoin is absorbed in healthy volunteers. DESIGN: This single-center, open-label crossover pilot study compared rectal absorption with intravenous administration of phenytoin injectable solution (7 mg/kg) in healthy volunteers. Twelve serial blood samples were taken from each volunteer beginning at time zero until 36 hours following administration. These were analyzed for presence of phenytoin by immunoassay. SETTING: The study took place at St. Paul's Hospital, a tertiary care center. PARTICIPANTS: Funding permitted for a sample size of five healthy participants, two men and three women, aged 21-45 years. Selection was by volunteer sample. Inclusion criteria were as follows: no known medical conditions, not receiving medication, no history of adverse drug reactions or allergies, not known to be pregnant, and normal liver function as determined per study protocol. MAIN OUTCOME MEASURES: Signs of absorption as indicated by presence of phenytoin in blood samples, maximum concentrations (Cmax), time to Cmax (tmax), AUC, and apparent bioavailability. RESULTS: Maximum mean concentrations of 2.4+/-1.1 mg/L (mean +/- SD) following rectal administration and 11.2+/-1.6 mg/L following intravenous administration were achieved during the first one to two hours (tmax in both treatment arms). Mean apparent bioavailability of the rectally administered phenytoin was 24.4+/-13.4% (15.8 44.1%). CONCLUSIONS: Results from this pilot study demonstrate that rectal absorption of phenytoin begins within 30 minutes following single-dose administration and was reported by four out of five volunteers to be the preferred route. Further studies are required before extrapolation can be made to the patient population. PMID- 10466904 TI - Fatal status epilepticus associated with olanzapine therapy. AB - OBJECTIVE: To report a case of fatal status epilepticus in a patient using olanzapine with no known underlying cause or predisposing factor for seizure. CASE SUMMARY: A 41-year-old white woman developed witnessed seizures at home that progressed to status epilepticus. She subsequently died from secondary rhabdomyolysis and disseminated intravascular coagulation. She had been taking olanzapine for five months prior to the event. No other toxic, metabolic, or anatomic abnormalities were identified pre- or postmortem to explain the seizures. Her seizures were a probable adverse drug reaction based on the Naranjo scale. DISCUSSION: This is the first case of fatal status epilepticus described that has been associated with the use of olanzapine. The pharmacodynamics of olanzapine are similar to those of clozapine, which has been described to induce seizures in 1-4% of patients. It is possible that this patient may have suffered seizures due to a similar effect. Alternate explanations include neuroleptic malignant syndrome and alcohol or benzodiazepine withdrawal seizures, although her clinical history does not suggest these etiologies. CONCLUSIONS: Although olanzapine has infrequently been associated with seizures in premarketing studies, its potential to induce them exists. Postmarketing surveillance should continue to determine how significant this effect may be. PMID- 10466905 TI - Potential fluconazole-induced carbamazepine toxicity. AB - OBJECTIVE: To report a case of carbamazepine toxicity resulting from a drug interaction with fluconazole, and to review the possible mechanisms of this interaction. DATA SOURCES: Medical record review. DATA SYNTHESIS: A 33-year-old white man with a history of mental retardation and seizures experienced stupor due to carbamazepine toxicity after fluconazole was initiated. The patient had been taking carbamazepine for several years and maintained stable therapeutic concentrations. He started fluconazole therapy after developing a rash presumably due to candidiasis. After admission to the hospital for carbamazepine toxicity, both fluconazole and carbamazepine were withheld and the patient returned to his normal baseline mental status once the carbamazepine concentration declined to a therapeutic range. Carbamazepine was restarted and the patient experienced no further adverse events. Carbamazepine is metabolized by the cytochrome P450 3A4 isoenzyme. Fluconazole is renally excreted but has been noted to inhibit CYP3A4. Fluconazole has also been noted to increase phenytoin concentrations. CONCLUSIONS: Fluconazole may cause carbamazepine toxicity presumably by inhibiting the cytochrome P450 3A4 isoenzyme. PMID- 10466906 TI - Levofloxacin-induced bilateral Achilles tendonitis. AB - OBJECTIVE: To report a case of possible levofloxacin-induced bilateral Achilles tendonitis. CASE SUMMARY: An 83-year-old white woman presented to her physician with five days of hemoptysis. She was diagnosed with right lower-lobe pneumonia based on chest X-ray, and levofloxacin 500 mg/d po for 10 days was prescribed. Three days into treatment she began having a variety of adverse effects, including severe nausea, constipation, stomach cramps, and dizziness. Signs of tendonitis began three days after treatment and peaked four days after completion of therapy. Two weeks later, she was treated by her podiatrist with an ankle immobilizer and rest. At her three-week follow-up, she had marked improvement in her pain and bruising; however, her symptoms had not completely resolved. DISCUSSION: Tendonitis and tendon rupture are rare adverse effects of fluoroquinolone antibiotics; there are no reports in the literature of levofloxacin-induced tendonitis. As newer fluoroquinolones become available, the postmarketing studies will become increasingly important to capture the data on rare but serious adverse effects not discovered in the premarketing trials. CONCLUSIONS: To our knowledge, this is the first reported case of tendonitis caused by levofloxacin reported in the literature. Reports have been made, however, to the manufacturer via postmarketing surveillance. As more people are treated with newer fluoroquinolones, the clinical incidence of tendon rupture with these agents may become clearer. PMID- 10466908 TI - Acute renal failure after large doses of intravenous immune globulin. AB - OBJECTIVE: To describe a case of acute renal failure after high-dose intravenous immune globulin (IVIG) therapy and the measures undertaken to prevent this complication during subsequent administration. CASE SUMMARY: A 54-year-old white man with valvular cardiomyopathy was receiving large doses (2 g/kg/mo) of IVIG in order to attenuate his immune system in preparation for a heart transplant. After his first infusion, he had to be rehospitalized for nausea, vomiting, fever, chills, and acute renal failure (serum creatinine [Scr] peak 8.4 mg/dL, baseline 1.0 mg/dL). His second infusion produced similar complications. Sandoglobulin 100 mL/h (172 g; 10% solution prepared with sterile water) was used on both occasions, and the large sucrose load (1.67 g sucrose/g protein) was suspected to be the causative agent. Upon switching to Polygam (170 g; 10% solution prepared with sterile water), a glucose-containing product which only has 0.4 g glucose/g protein, and infusing it at half of the Sandoglobulin rate (50 mL/h), the patient was able to tolerate the infusion without complications (Scr and blood urea nitrogen unchanged). DISCUSSION: Stabilizing agents such as sucrose, maltose, and glucose are added to IVIG preparations to help reduce immunoglobulin aggregation. These aggregates are associated with some of the more serious adverse effects of IVIG administration. When large doses of IVIG are used, the stabilizing agents can induce an osmotic nephrosis due to the large solute load. A review of the previous literature on IVIG-induced renal failure is provided, as well as the differences in the various IVIG formulations. Also, general guidelines are offered to prevent this complication. CONCLUSIONS: Large doses of Sandoglobulin (400-2000 mg/kg) have been associated with acute renal failure due to the large sucrose load. By taking certain precautions, especially in high-risk patients, this uncommon, but serious, adverse effect can be avoided. PMID- 10466909 TI - High-dose methotrexate-doxycycline interaction. AB - OBJECTIVE: To report a case in which an interaction occurred between doxycycline and high-dose methotrexate (HD-MTX) and discuss its clinical, quality of life, and economic repercussions. CASE SUMMARY: A 17-year-old girl diagnosed with high degree osteosarcoma in her left femur was admitted to our hospital to receive her eleventh postoperative cycle of HD-MTX, according to the clinical protocol approved at our hospital. Simultaneously, due to a palpebral abscess in her left eye, the patient received systemic treatment with doxycycline 100 mg every 12 hours. As in previous cycles, pharmacokinetic monitoring of methotrexate (MTX) plasma concentrations was performed to improve leucovorin calcium rescue. The biological half-life 12 and 24 hours after infusion showed that the patient was at high risk of intoxication. She then developed hematologic as well as gastrointestinal toxicity and remained hospitalized for 11 days. The patient was readmitted 48 hours after hospital discharge because of a febrile episode that required an additional hospital stay of 12 days, and the twelfth cycle of HD-MTX had to be postponed 18 days because of the results of the previous cycle. DISCUSSION: Significant differences (p < 0.05) were observed in the eleventh HD MTX cycle for plasma concentrations and pharmacokinetic parameters compared with the previous 10 cycles. The interaction resulted in clinical, quality of life, and economic repercussions. CONCLUSIONS: The potential interaction between MTX and doxycycline may cause pharmacokinetic changes, and its clinical repercussions on the quality of life of the patient and associated costs should be considered. PMID- 10466907 TI - Concomitant digoxin toxicity and warfarin interaction in a patient receiving clarithromycin. AB - OBJECTIVE: To report a case of a clarithromycin-associated warfarin interaction and digoxin toxicity in a patient. CASE SUMMARY: A 72-year-old white woman with chronic atrial fibrillation receiving long-standing therapy with digoxin 0.25 mg/d and warfarin 22.5 mg/wk was prescribed clarithromycin 500 mg three times daily for eradication of Helicobacter pylori. The patient presented to the emergency department with gastrointestinal symptoms, weakness, dizziness, and visual changes 12 days after initiation of clarithromycin. Laboratory results revealed a serum digoxin concentration of 4.6 ng/mL (normal 1.0-2.6) and an international normalized ratio of 7.3 (2.0-3.0). Digoxin, warfarin, and clarithromycin were discontinued and the patient was admitted to the hospital for treatment to resolve the symptoms and to return laboratory values to a safe range. Reduced dosages of digoxin (0.125 mg/d) and warfarin (17.5 mg/wk) were restarted on day 7 of hospitalization. The patient was discharged on day 11 in good condition. DISCUSSION: Several reports of clarithromycin-induced drug interactions with digoxin and with warfarin have been published. Previously, case reports of macrolide-associated interactions mainly involved erythromycin, but more recently have implicated clarithromycin. The interaction between clarithromycin and warfarin is thought to occur from an inhibition of the cytochrome P450 drug metabolizing system. Clarithromycin is thought to cause digoxin toxicity by an alteration of the digoxin-metabolizing gut flora, thereby causing an increase in the digoxin concentration in susceptible individuals. Drug interactions can occur by different mechanisms in the same patient. CONCLUSIONS: Potential drug interactions can occur between commonly prescribed medications. It is important to monitor patients for symptoms and alterations in laboratory values to prevent not only serious complications, but also unnecessary hospitalizations. PMID- 10466910 TI - Estrogen for dementia-related aggression in elderly men. AB - OBJECTIVE: To report the use of conjugated estrogens for aggression in two elderly men with dementia and to review the available literature relating to estrogen use for aggression in the elderly. CASE SUMMARIES: Case 1. A 78-year-old white man had probable Alzheimer dementia and aggression (verbal and physical) that was nonresponsive to antipsychotic, antidepressant, and mood stabilizer therapy. However, conjugated estrogens 1.875 mg/d reduced his physical aggression, as evidenced by a 75% decline in aggressive episodes, per 24-hour nursing reports and progress notes. Case 2. A 78-year-old African-American man with vascular dementia, physical aggression, and sexual inappropriateness was unsuccessfully managed with several antipsychotic and benzodiazepine trials. He did, however, exhibit a decline in physical and sexual aggression by 80%, as well as a 55% reduction in sexual comments, after daily treatment with conjugated estrogens 0.625 mg. Estrogen was well tolerated by both patients. DATA SOURCES: MEDLINE (January 1966-May 1998) and PsychLit (January 1974-May 1998) database searches were performed to identify case reports, letters, or clinical trials discussing estrogen use in aggressive elderly patients. CONCLUSIONS: As with previous reports, these cases suggest that conjugated estrogens may be used to reduce physical and sexual aggression associated with dementia in elderly men. Additional study is needed to determine the exact role of this alternative pharmacologic intervention. PMID- 10466911 TI - Donepezil overdose: a tenfold dosing error. AB - OBJECTIVE: To report toxicity resulting from donepezil administration following a tenfold dosing error. CASE SUMMARY: A 79-year-old white nursing home patient with a history of Alzheimer disease and hypertension was inadvertently given 50 mg of donepezil instead of her usual 5-mg dose. She presented to the emergency department with nausea, vomiting, and persistent bradycardia (HR in the 40s). Routine laboratory studies were all within normal limits. Basilar rales were noted five hours after arrival. She was treated with atropine 0.2 mg as needed for bradycardia (HR <50 beats/min); a total of 3.0 mg was administered over 18 hours. Each bolus kept her HR >60 beats/min for one-half to two hours. No further vomiting or evidence of pulmonary edema occurred after her initial episodes. She returned to baseline by day 2 (HR in the 70s) and was returned to the nursing home. DISCUSSION: Donepezil is a centrally acting, reversible cholinesterase inhibitor that is used in the treatment of Alzheimer disease. Donepezil is highly specific for neural acetylcholinesterases, preferentially binding acetylcholinesterase by greater than three orders of magnitude over butyrylcholinesterases. This specificity minimizes peripheral adverse effects at therapeutic doses. Our patient mainly experienced bradycardia and had minimal secretory effects compared with what is usually seen with nonspecific cholinesterase inhibition. Medication errors like the one that produced this overdose are a common but preventable cause of morbidity in healthcare facilities. CONCLUSIONS: A tenfold dosing error caused donepezil toxicity. The main effect of this overdose was bradycardia, which responded to atropine therapy. PMID- 10466912 TI - Clinical presentation, diagnosis, and pharmacotherapy of patients with primary brain tumors. AB - OBJECTIVE: To briefly review the clinical presentation and diagnosis of patients with primary brain tumors, followed by an in-depth survey of the pertinent pharmacotherapy. DATA SOURCES: A detailed search of the neurologic, neurosurgical, and oncologic literature for basic science research, clinical studies, and review articles related to chemotherapy and pharmacotherapy of primary brain tumors. STUDY SELECTION: Relevant studies on tissue culture systems, animals, and humans examining the mechanisms of action, pharmacokinetics, clinical pharmacology, and treatment results of chemotherapeutic agents for primary brain tumors. In addition, studies of pharmacologic agents administered for supportive care and symptom control are reviewed. DATA SYNTHESIS: Primary brain tumors derive from cells within the intracranial cavity and generally present with headache, seizure activity, cognitive changes, and weakness. They are diagnosed most efficiently with magnetic resonance imaging. After diagnosis, the most common supportive medications include corticosteroids, gastric acid inhibitors, and anticonvulsants. Chemotherapy is adjunctive treatment for patients with malignant tumors and selected recurrent or progressive benign neoplasms. In general, the most effective chemotherapeutic drugs are alkylating agents such as the nitrosoureas, procarbazine, cisplatin, and carboplatin. Other agents used include cyclophosphamide, methotrexate, vincristine, and etoposide. Angiogenesis inhibitors and gene therapy comprise some of the novel therapeutic strategies under investigation. CONCLUSIONS: The efficacy of chemotherapy for primary brain tumors remains modest. Novel agents must be discovered that are more specific and attack tumor cells at the molecular level of tumorigenesis. Furthermore, strategies must be developed to counteract the pervasive problem of brain tumor chemoresistance. PMID- 10466913 TI - Tetravalent rotavirus vaccine. AB - OBJECTIVE: To review the clinical efficacy, safety, and pharmacoeconomic data about the use of rhesus-human reassortant rotavirus tetravalent vaccine (RRV-TV) in infants and children. DATA SOURCES: A MEDLINE search (January 1990-December 1998) was conducted to identify all publications on the RRV-TV vaccine including pharmacology, clinical trials, adverse effects, and pharmacoeconomics in infants and children. Bibliographies of articles were also used. STUDY SELECTION: All randomized and placebo-controlled clinical efficacy trials were reviewed. Additionally, pharmacoeconomic studies focusing on the potential impact on healthcare costs were chosen for review. DATA SYNTHESIS: Rotavirus-induced gastroenteritis is a significant problem in developed and developing countries. Various forms of a rotavirus vaccine have been studied worldwide. The tetravalent vaccine appears to have similar efficacy in developed and developing countries. It seems to be most effective against the most severe forms of gastroenteritis, with an 80% overall efficacy rate. This vaccine is well tolerated; the most common adverse effect is fever after the first dose. Pharmacoeconomic studies indicate that although the vaccine may be only moderately effective against less severe gastroenteritis, over $1 billion annually could potentially be saved in the US with its universal use. CONCLUSIONS: The new rotavirus vaccine is effective in preventing and reducing the incidence of rotavirus-induced gastroenteritis. The morbidity, mortality, and healthcare costs from this disease may be reduced if this vaccine is provided to children worldwide. PMID- 10466914 TI - Pharmacologic management of Alzheimer disease part III: nonsteroidal antiinflammatory drugs--emerging protective evidence? AB - OBJECTIVE: To provide information about research evaluating the role of nonsteroidal antiinflammatory drugs (NSAIDs) in the prevention, or delay in the onset of, Alzheimer disease (AD). DATA SOURCES: Studies, review articles, and editorials identified from MEDLINE searches (January 1990-December 1996) and bibliographies of identified articles. The addendum lists articles from 1996 to June 1999. STUDY SELECTION: Studies, review articles, and editorials addressing NSAIDs and AD pharmacotherapy research. DATA EXTRACTION: Pertinent information was selected and the data synthesized into a review format. DATA SYNTHESIS: AD is a complex disorder and there are numerous factors involved in the process. The pathology of AD is characterized by the development of amyloid plaques and neurofibrillary tangles. In addition, more than 40 immunoprotective proteins that are not unique to AD are found at autopsy that are normally absent, or present in very low concentrations, in normal brain. These findings suggest that AD may involve an inflammatory process. Preliminary results from studies investigating the incidence and onset of AD in patients with arthritis who have taken NSAIDs suggest that NSAIDs may have a protective effect. Studies evaluating the possible association between arthritis, NSAIDs, and AD are reviewed. CONCLUSIONS: Preliminary evidence suggests that NSAIDs may have a protective effect against the development of AD. Further prospective, double-blind, placebo-controlled studies are needed to determine the role of NSAIDs in AD. These dose-finding studies should focus on specific agents and identify the dosage and duration of therapy necessary for a protective or therapeutic effect. Additionally, not all elderly patients are candidates for NSAIDs. Determining the definitive mechanism of action of NSAIDs in AD may suggest alternative agents that have similar pharmacologic activity, but are associated with fewer adverse effects. PMID- 10466915 TI - An interdisciplinary mock trial involving pharmacy, law, and ethics. AB - OBJECTIVE: To describe an innovative, interdisciplinary approach to illustrate the relationship between healthcare ethics and law. METHODS: A mock trial was created for students enrolled in the Samford University McWhorter School of Pharmacy and Department of Paralegal Education. The trial served as the starting point to discuss confidentiality in health care in general and pharmacy in particular. Students from both programs served on the jury and rendered a verdict after the case had been presented. The Alabama statute concerning exceptions to confidentiality is reviewed. The students' assignment and lessons learned are also described. SUMMARY: Students thoroughly enjoyed this method of teaching and learning. The mock trial provided an interesting way to exemplify the often complex relationship between healthcare ethics and law. PMID- 10466917 TI - Lamotrigine--an effective mood stabilizer? AB - OBJECTIVE: To review the literature regarding the use of lamotrigine as a mood stabilizer, and to discuss its efficacy in treating this condition. DATA SOURCES: Data were obtained from MEDLINE, Micromedex, and Cochrane collaboration searches from January 1985 to July 1998. DATA SUMMARY: There are insufficient data to confirm that lamotrigine is an effective mood stabilizer. There are no controlled studies, and the current evidence is from case studies and open trials. Furthermore, only one study shows any evidence of effectiveness in the manic phase, although this may be because the data tend to relate to a treatment refractory population. CONCLUSIONS: From the current evidence, lamotrigine cannot be recommended as a mood stabilizer except when conventional therapies have failed. PMID- 10466916 TI - Hyperlipidemia associated with protease inhibitor therapy. AB - OBJECTIVE: To report a case of extreme hyperlipidemia associated with protease inhibitor-based antiretroviral therapy and review the relevant literature concerning lipid abnormalities with HIV infection and antiretroviral therapy. CASE SUMMARY: A 35-year-old HIV-infected man developed a serum cholesterol of 1472 mg/dL and fasting serum triglycerides of 8660 mg/dL after initiation of antiretroviral therapy consisting of ritonavir, saquinavir, nevirapine, and didanosine. All other medications had been stable during this time period and the abnormality resolved after discontinuation of antiretroviral therapy and initiation of lipid-lowering therapy. The elevated cholesterol and triglyceride concentrations did not recur when therapy was reinstituted with nelfinavir, saquinavir, nevirapine, and didanosine. The hyperlipidemia then was attributed to ritonavir. DISCUSSION: Lipid abnormalities are common in patients with HIV infection and usually consist of hypocholesterolemia and moderate hypertriglyceridemia. Hypercholesterolemia and hypertriglyceridemia have been reported with ritonavir and, less commonly, with other currently available protease inhibitors. Some cases of ritonavir-associated hyperlipidemia have been extreme. Although an association between hyperlipidemia and clinical consequences such as pancreatitis and atherosclerotic disease has not been well described with protease inhibitor therapy, pancreatitis is common in HIV-infected patients. It is possible that in some cases, protease inhibitor-induced hypertriglyceridemia may contribute to the development of pancreatitis. CONCLUSIONS: Optimal management of lipid abnormalities in HIV-infected patients is controversial. The potential benefit of reducing the incidence of pancreatitis and atherosclerotic events must be weighed against the risk of intolerance, toxicity, and drug interactions. PMID- 10466918 TI - Interaction between ciprofloxacin and rifampin. AB - OBJECTIVE: To discuss the necessity of dose adjustment for ciprofloxacin or rifampin during their concurrent use. DATA SOURCES: A MEDLINE search (1966 December 1998) was completed using key terms rifampin and fluoroquinolone. English-language journals were considered. DATA SYNTHESIS: Studies in elderly patients after 14 days of therapy with oral ciprofloxacin and rifampin did not demonstrate significant differences in the pharmacokinetics of ciprofloxacin as compared with those in patients receiving ciprofloxacin alone. Similar results were found in intravenous drug abusers. In comparison to the pharmacokinetics of both ciprofloxacin and rifampin when given alone, the serum bactericidal activity of rifampin, when given with ciprofloxacin in healthy elderly volunteers, was reduced but still evident. Ciprofloxacin pharmacodynamics were not significantly altered. Serum bacterial titers of ciprofloxacin and pefloxacin increased twofold when given with rifampin, although their clinical significance is unknown. RECOMMENDATION: No strong evidence of a significant interaction exists to support dose adjustment for ciprofloxacin or rifampin during their concurrent use. PMID- 10466920 TI - Effect of short-term rifampin on stable cyclosporine concentrations. PMID- 10466919 TI - Possible interaction between nefazodone and pravastatin. PMID- 10466921 TI - ACE inhibitor switch in an indigent community clinic. PMID- 10466922 TI - Lack of toxicity in a cladribine extravasation. PMID- 10466923 TI - Paclitaxel compatibility with ethylene vinyl acetate bags. PMID- 10466924 TI - Studies on heat shock proteins in sea urchin development. AB - Work on stress proteins in sea urchin embryos carried out over the last 20 years is reviewed and the following major results are described. Entire sea urchin embryos, if subjected to a rise in temperature at any postblastular stage undergo a wave of heat shock protein (hsp) synthesis and survive. If subjected to the same rise between fertilization and blastula formation, they are not yet able to synthesize hsp and die. Four clones coding for the major hsp, hsp70, have been isolated and sequenced; evidence for the existence of a heat shock factor has been provided, and a mechanism for the developmental regulation of hsp synthesis discussed. Intraembryonic and intracellular hsp location has been described; and a mechanism for achievement of thermotolerance proposed. A chaperonine role for a constitutive mitochondrial hsp56 has been suggested, as well as a role for the constitutive hsp70 in cell division. Heat shock, if preceded by 12-O tetradecanoylphorbol-12-acetate (TPA) treatment causes apoptosis. PMID- 10466925 TI - Generation and characterization of a monoclonal antibody, mH1, raised against mitotic HeLa cells. AB - Hybridoma cell lines were prepared from spleen cells of mouse immunized with mitotic HeLa cells. A monoclonal antibody (mH1), which intensively reacted with cleavage furrows of dividing HeLa cells in immunofluorescence, was obtained. In interphase, this antibody diffusely stained whole HeLa cells. Immunoelectron microscopy showed that mH1 antigens were localized at microvillus projections at the surface of dividing HeLa cells, but definite localization of that antigen was not observed in interphasic cells. Immunoblot analysis showed that mH1 is reactive to 42-kDa and 130-kDa components. Further, the 42-kDa component was identified as a gamma-actin homolog by N-terminal amino acid sequence analysis. PMID- 10466926 TI - Endoderm differentiation and inductive effect of activin-treated ectoderm in Xenopus. AB - When presumptive ectoderm is treated with high concentrations of activin A, it mainly differentiates into axial mesoderm (notochord, muscle) in Xenopus and into yolk-rich endodermal cells in newt (Cynops pyrrhogaster). Xenopus ectoderm consists of multiple layers, different from the single layer of Cynops ectoderm. This multilayer structure of Xenopus ectoderm may prevent complete treatment of activin A and subsequent whole differentiation into endoderm. In the present study, therefore, Xenopus ectoderm was separated into an outer layer and an inner layer, which were individually treated with a high concentration of activin A (100 ng/mL). Then the differentiation and inductive activity of these ectodermal cells were examined in explantation and transplantation experiments. In isolation culture, ectoderm treated with activin A formed endoderm. Ectodermal and mesodermal tissues were seldom found in these explants. The activin-treated ectoderm induced axial mesoderm and neural tissues, and differentiated into endoderm when it was sandwiched between two sheets of ectoderm or was transplanted into the ventral marginal zone of other blastulae. These findings suggest that Xenopus ectoderm treated with a high concentration of activin A forms endoderm and mimics the properties of the organizer as in Cynops. PMID- 10466927 TI - Newt RAD51: cloning of cDNA and analysis of gene expression during spermatogenesis. AB - A cDNA encoding a newt homolog of Escherichia coli RecA and yeast RAD51 from a testis cDNA library was isolated. The newt RAD51 (nRAD51) cDNA predicted a 337 amino acid protein with a 95-96% amino acid identity to Xenopus and mammalian RAD51. Northern blot analysis showed that nRAD51 mRNA, 1.7 kb in length, was expressed strongly in the testis and ovary, but weakly in the liver, kidney and brain. In situ hybridization revealed that expression of nRAD51 mRNA was barely observed in primary spermatogonia (one cell in a cyst) and early secondary spermatogonia (two to four cells in a cyst), but increased in late secondary spermatogonia (> or =eight cells in a cyst), reaching a maximum level in leptotene-zygotene spermatocytes, and thereafter declined. These results suggest that nRAD51 is involved in mitotic recombination in spermatogonia as well as in meiotic recombination in spermatocytes. PMID- 10466928 TI - A calcium-binding motif in SPARC/osteonectin inhibits chordomesoderm cell migration during Xenopus laevis gastrulation: evidence of counter-adhesive activity in vivo. AB - Secreted protein, acidic, rich in cysteine (SPARC) is a Ca2+-binding, counter adhesive, extracellular glycoprotein associated with major morphogenic events and tissue remodeling in vertebrates. In Xenopus laevis embryos, SPARC is expressed first by dorsal mesoderm cells at the end of gastrulation and undergoes complex, rapid changes in its pattern of expression during early organogenesis. Another study has reported that precocious expression of SPARC by injection of native protein into the blastocoele cavity of pregastrula embryos leads to a concentration-dependent reduction in anterior development. Thus, normal development requires that the timing, spatial distribution, and/or levels of SPARC be regulated precisely. In a previous study, we demonstrated that injection of a synthetic peptide corresponding to the C-terminal, Ca2+-binding, EF-hand domain of SPARC (peptide 4.2) mimicked the effects of native SPARC. In the present investigation, peptide 4.2 was used to examine the cellular and molecular bases of the phenotypes generated by the aberrant presence of SPARC. Exposure of late blastula embryos to LiCl also generated a concentration-dependent reduction in anterior development; therefore, injections of LiCl were carried out in parallel to highlight the unique effects of peptide 4.2 on early development. At concentrations that caused a similar loss in anterior development (60-100 ng peptide 4.2 or 0.25-0.4 microg LiCl), LiCl had a greater inhibitory effect on the initial rate of chordomesoderm cell involution, in comparison with peptide 4.2. However, as gastrulation progressed, peptide 4.2 had a greater inhibitory effect on prospective head mesoderm migration than that seen in the presence of LiCl. Moreover, peptide 4.2 and LiCl had distinct influences on the expression pattern of dorso-anterior markers at the neural and tail-bud stages of development. Scanning electron microscopy showed that peptide 4.2 inhibited spreading of migrating cells at the leading edge of the involuting chordomesoderm. While still in close proximity to the blastocoele roof, many of the cells appeared rounded and lacked lamellipodia and filopodia extended in the direction of migration. In contrast, LiCl had no effect on the spreading or shape of involuting cells. These data are the first evidence of a counter-adhesive activity for peptide 4.2 in vivo, an activity demonstrated for both native SPARC and peptide 4.2 in vitro. PMID- 10466929 TI - A maternal RNA encoding smad1/5 is segregated to animal blastomeres during ascidian development. AB - Expressed Sequence Tag (EST) research on the ascidian Halocynthia roretzi revealed that Hrsmad1/5, a homolog of smad genes, is expressed in H. roretzi eggs. A comparison of amino acid sequences of smad family members showed that the isolated clone was a homolog of smad1 and smad5 of vertebrates. A molecular phylogenetic analysis showed that Hrsmad1/5 was separated from the common ancestor with the group containing smad1 and smad5. A northern blot analysis showed that transcript of Hrsmad1/5 was abundant in the fertilized egg. The amount of the transcript remained constant until the gastrulae and then rapidly decreased at the neurulae. The spatial expression of Hrsmad1/5 was investigated by means of whole-mount in situ hybridization. Maternal transcripts of Hrsmad1/5 were detected in the entire fertilized egg. The signals were localized preferentially to the animal blastomeres of the 8-, 16-, 32- and 64-cell stages. The zygotic expression of Hrsmad1/5 started in prospective epidermal blastomeres in the animal hemisphere at the 64-cell stage but not in cells of the central nervous system, and it decreased rapidly around the neural-plate stage. At the tail-bud stage, signals were detected broadly all through the trunk and in a small part of the epidermis in the tail region. This is the first report of a maternal RNA that preferentially accumulates in the animal hemisphere in early ascidian embryos. Animal blastomeres of ascidian embryos differentiate mainly into epidermis in a cell-autonomous manner and partly differentiate into neural tissues by induction. The Hrsmad1/5 gene may play a role in the signal transduction process in epidermal precursor cells of ascidian embryos. PMID- 10466930 TI - Lens formation by pigmented epithelial cell reaggregate from dorsal iris implanted into limb blastema in the adult newt. AB - In newt lens regeneration, the dorsal iris has lens forming ability and the ventral iris has no such capability, whereas there is no difference in the morphological criteria. To investigate the real aspects of this characteristic lens regeneration in the newt at the cellular level, a useful model system was constructed by transplanting the dorsal and ventral reaggregate derived from singly dissociated pigmented epithelial cells of the iris into the blastema of the forelimb in the newt. The lens was formed from the dorsal reaggregate with high efficiency, but not from the ventral one. No lens formation was observed in the implantation of the reaggregate into the tissue of the intact limbs. In detailed examination of the process of lens formation from the reaggregate, it was shown that tubular formation was the first step in the rearrangement of cells within the reaggregate. This was followed by depigmentation, vesicle formation with active cell growth, and the final step was lens fiber formation by transdifferentiation of epithelial cells composing the lens vesicle. The process was almost the same as in situ lens regeneration except the reconstitution of the two-layered epithelial structure was embodied as flattened tubular formation in the first step. The present study made it possible for the first time to examine lens forming ability in the reaggregate mixed with dorsal and ventral cells, because the formation of a reaggregate was started from singly dissociated cells of the dorsal and ventral cells of the iris. Mixed reaggregate experiments indicated that the existence of the dorsal cells in a cluster within the reaggregate is important in lens formation, and ventral cells showed an inhibitory effect on the formation. The present study demonstrated that the limb system thus constructed was effective for the analysis of lens formation at the cellular level and made it possible to examine the role of dorsal and ventral cells in lens regeneration. PMID- 10466931 TI - Absence of furrowing activity following regional cortical tension reduction in sand dollar blastomere and fertilized egg fragment surfaces. AB - The purpose of the present investigation was to test experimentally the possibility that division mechanism establishment at the equator of sand dollar eggs may be a consequence of cortical tension gradients between the equator and the poles. Cytochalasin has been shown to decrease tension at the sea urchin egg surface. The concave ends of cytochalasin D-containing agarose cylinders were held against regions of the surface of Echinarachnius parma blastomeres and enucleated fertilized egg fragments. The ability to interfere with normal furrowing activity was used as a biological indicator of the effectiveness of cytochalasin. When agarose containing 2 microg/mL cytochalasin contacted the equatorial region of the blastomeres resulting from the first cleavage, or the equatorial surfaces of nucleated fertilized egg halves, furrowing was blocked, stalled or delayed, indicating that the concentration of cytochalasin was effective. When the same concentration of cytochalasin was applied to the poles, the cells and nucleated fertilized egg fragments divided in the same way as the controls, indicating that the effectiveness of the cytochalasin did not spread from the poles to the equator and that bisection did not interfere with the division of nucleated fertilized egg fragments. When the same concentration of cytochalasin was applied to diametrically opposed surfaces of enucleated, spherical egg fragments, there was no evidence of furrowing activity between the areas that contacted the cytochalasin or in any other part of the surface. Because of the tension-reducing effect of cytochalasin, a tension gradient existed between the regions affected and unaffected by cytochalasin. The results strongly suggest that establishment of the division mechanism by simple gradients of tension at the surface is unlikely. PMID- 10466932 TI - Duration of competence and inducing capacity of blastomeres in notochord induction during ascidian embryogenesis. AB - Notochord cells in ascidian embryos are formed by the inducing action of cells of presumptive endoderm, as well as neighboring presumptive notochord, at the 32 cell stage. Studies of the timing of induction using recombinations of isolated blastomeres have suggested that notochord induction must be initiated before the decompaction of blastomeres at the 32-cell stage and is completed by the 64-cell stage. However, it is not yet clear how the duration of notochord induction is strictly limited. In the present paper, the aim was to determine in detail when the presumptive notochord blastomeres lost their competence to respond, and when the presumptive endoderm blastomeres produced inducing signals for the notochord. Presumptive notochord blastomeres and presumptive endoderm blastomeres were isolated from early 32-cell embryos, and were heterochronously recombined at various stages ranging from the early 32-cell stage to the 64-cell stage. Presumptive notochord blastomeres could respond to inductive signals at the early 32-cell stage, and started to lose their responsiveness at the decompaction stage. By contrast, the presumptive endoderm blastomeres persisted in their inducing capacity even at the 64-cell stage. These observations suggest that the loss of competence in presumptive notochord blastomeres limits the duration of notochord induction in intact ascidian embryos. PMID- 10466933 TI - Vitelline-coat lysin: comparison of lysins among three species in the genus Tegula. AB - Vitelline-coat lysins from two species of marine mollusc of the genus Tegula, Tegula lischkei and Tegula sp., were purified and their properties compared with those of Tegula pfeifferi. Cross-reaction tests among these three species proved that the lysin action on the vitelline coat was species specific. Each of the lysins from these three species is a single, basic polypeptide with a molecular weight of 16000-17000 and an isoelectric point of pH 10.5. All of them share a common antibody recognition site(s) for the anti-T. pfeifferi lysin antibody. Their amino acid sequences were analyzed using intact lysins and peptides separated by reverse phase high-performance liquid chromatography after V8 proteinase digestion. The amino acid sequences of the N-terminus (Nos 1-66) from the three species showed 98% homology, and those of the C-terminus (Nos 91-118) showed 89% homology. It was concluded that the species specificity of the vitelline coat lysin appears to be determined by a sequence of 24 residues (Nos 67-90) in the middle region of the molecule. PMID- 10466934 TI - cDNA cloning of a stage-specific gene expressed during HCG-induced spermatogenesis in the Japanese eel. AB - A single injection of human chorionic gonadotropin (HCG) can induce complete spermatogenesis in immature Japanese eel (Anguilla japonica) testes consisting of only premitotic spermatogonia. Proliferation of spermatogonia, meiosis and spermiogenesis begin on 3, 12 and 18 days after HCG injection, respectively. To isolate the genes responsible for regulating the initiation of meiosis, differential mRNA display using poly (A)+ RNA extracted from testes of eels at different times after HCG treatment was carried out. Five cDNA clones in which expression was initiated before the onset of meiosis were obtained. Northern blot analysis showed that one clone, which encoded activin betaB subunit, was expressed in the initial phase of spermatogenesis (1-6 days after HCG treatment), in agreement with the previous suggestion that activin B induces the initiation of spermatogenesis in the Japanese eel. The remaining four were expressed in the testes during the following time frames: 3-18 days (two clones), 6-18 days (one clone) and 9-18 days (one clone) after HCG treatment. One of the two clones expressed on day 3 exhibited strong expression on days 12 and 15, just at the initiation period of meiosis. This clone was selected as a candidate gene responsible for initiating meiosis, and its full-length cDNA isolated. The cDNA contained an open reading frame of 1571 nucleotides encoding a protein of 260 amino acid residues, which showed high homology with the proliferating cell nuclear antigen (PCNA) of human, mouse and Xenopus. Northern blot analysis using eel PCNA cDNA showed that a 1.6 kb transcript first appeared on day 3 and became abundant, reaching maximum levels on days 12-15. In situ hybridization analysis revealed that PCNA mRNA was expressed strongly in late type B spermatogonia before the sixth mitotic division. It has already been shown that spermatogonia have a regulatory point to enter meiosis between the fifth and sixth mitotic division. The coincidence of PCNA expression and this regulatory point suggests an involvement of PCNA in the progression of mitotic germ cells into meiosis during HCG-induced spermatogenesis in the eel. PMID- 10466935 TI - Studies on fertilization of the teleost. II. Nuclear behavior and changes in histone H1 kinase. AB - In order to understand the dynamic responses of gamete nuclei upon fertilization in the fish, Oryzias latipes, the relationship between changes in the activity of histone H1 kinase and nuclear behavior was examined during fertilization. Kinase activity rapidly decreased concomitant with the initiation of the propagative exocytosis of cortical alveoli following sperm attachment to the egg plasma membrane post-insemination (PI). Activity again increased 30 min PI. Similar changes in kinase activity, migration and syngamy of pronuclei, and subsequent cleavage were observed with aphidicolin or actinomycin D treatment, except that formation of abnormal metaphase chromosomes was retarded in aphidicolin-treated zygotes. Pretreatment of unfertilized eggs with cycloheximide or 6 dimethylaminopurine (6-DMAP) caused no nuclear changes. The activity of histone H1 kinase in these eggs rapidly declined following sperm penetration and exocytosis, but did not undergo subsequent increase in the presence of these inhibitors. In these eggs with low histone H1 kinase activity, the fertilization process from sperm penetration to syngamy occurred normally, but the pronuclear membrane did not break down and the chromosomes did not condense. The present data suggest that in fish eggs, DNA replication as well as the synthesis and phosphorylation of proteins, especially cyclin B, are required for normal formation of metaphase chromosomes at the first cleavage, but not for fertilization events from sperm penetration through to nuclear migration resulting in syngamy. PMID- 10466936 TI - Association of the sea urchin EGF-related peptide, EGIP-D, with fasciclin I related ECM proteins from the sea urchin Anthocidaris crassispina. AB - Exogastrula-inducing peptides (EGIP) of the sea urchin Anthocidaris crassispina are endogenous peptides related to epidermal growth factor (EGF), which induce exogastrulation in the embryo. Recently, a protein(s) from sea urchin embryos that binds to one of the EGIP, EGIP-D (EGIP-D-binding protein, EBP) was purified. The isolation and characterization of the cDNA clones for two EBP proteins (EBP alpha and EBP-beta) is reported. The two EBP proteins were highly similar in structure to each other; both possessed putative cell-binding sites and two repeated sequences characteristically seen in the insect neuronal cell adhesion protein, fasciclin I. The EBP showed similarity with other sea urchin proteins HLC-32, Bep1, and Bep4. It has been confirmed that bacterially expressed EBP proteins associate with EGIP-D as does native EBP, suggesting the interaction between EGF-related proteins and fasciclin I-related proteins. An EBP transcript of 1.4 kb was strongly expressed in immature ovaries but not in immature testes. A somewhat lower level of the transcript existed in unfertilized eggs and the amount gradually declined to an almost undetectable level by the pluteus stage. The EBP proteins were present throughout embryonic development at nearly constant levels. Although most of the proteins were distributed rather evenly in the cytoplasm, a small portion was detected on the apical surface of blastomeres and ectodermal cells, showing that EBP are components of the hyaline layer. PMID- 10466937 TI - Mitochondrial small ribosomal RNA is present on polar granules in early cleavage embryos of Drosophila melanogaster. AB - In Drosophila, formation of the germline progenitors, the pole cells, is induced by polar plasm localized in the posterior pole region of early embryos. The polar plasm contains polar granules, which act as a repository for the factors required for pole cell formation. It has been postulated that the factors are stored as mRNA and are later translated on polysomes attached to the surface of polar granules. Here, the identification of mitochondrial small ribosomal RNA (mtsrRNA) as a new component of polar granules is described. The mtsrRNA was enriched in the polar plasm of the embryos immediately after oviposition and remained in the polar plasm throughout the cleavage stage until pole cell formation. In situ hybridization at an ultrastructural level revealed that mtsrRNA was enriched on the surface of polar granules in cleavage embryos. Furthermore, the localization of mtsrRNA in the polar plasm depended on the normal function of oskar, vasa and tudor genes, which are all required for pole cell formation. The temporal and spatial distribution of mtsrRNA is essentially identical to that of mitochondrial large ribosomal RNA (mtlrRNA), which has been shown to be required for pole cell formation. Taken together, it is speculated that mtsrRNA and mtlrRNA are part of the translation machinery localized to polar granules, which is essential for pole cell formation. PMID- 10466938 TI - Mechanisms of 5-hydroxytryptamine-induced inhibition in the porcine myometrium. AB - The present experiments were designed to clarify the mechanisms of the inhibitory response of 5-hydroxytryptamine (5-HT) in the porcine uterine circular muscle. Inhibitory responses induced by 5-HT (1 nM-1 microM) were not affected by apamin (1 microM), charybdotoxin (100 nM) or glibenclamide (20 microM) but were significantly attenuated by 4-aminopyridine (3 mM) and tetraethylammonium (3 mM). Imidazole (100 microM) decreased but 3-isobutyl-1-methylxanthine (30 microM), milrinone (30 microM) and Ro 20-1724 (10 and 30 microM) potentiated the 5-HT induced inhibition. On the other hand, zaprinast (3-30 microM) had no significant effect on the inhibitory response of 5-HT. 5-HT caused a time (0-5 min)-and concentration (1 nM-1 microM)-dependent increase in the tissue cyclic AMP level, but had no effect on the tissue cyclic GMP level. A significant correlation (P < 0.05) was observed between the inhibition of contraction and tissue cyclic AMP level. The effect of 5-HT on contractile force and cytosolic Ca2+ level ([Ca2+]i) was investigated using fura-PE3-loaded myometrial strips. A low concentration of 5-HT (< or = 10 nM) inhibited the spontaneous contraction without changing the amplitude of the spontaneous [Ca2+]i increase, but a higher concentration of 5-HT (> or = 100 nM) decreased the resting [Ca2+]i and inhibited both the spontaneous [Ca2+]i increase and spontaneous contraction. High-K+ (50 mM) caused increases in muscle contractile force and [Ca2+]i. 5-HT concentrationdependently inhibited the high-K(+)-induced contraction (EC50, 45 nM) with only a small decrease in [Ca2+]i increase. Carbachol also caused increases in muscle contractile force and [Ca2+]i. 5-HT significantly decreased both the carbachol-induced contraction and [Ca2+]i increase, but was more potent at inhibition of contractile force than [Ca2+]i. In Ca(2+)-loaded myometrial strips, carbachol, but not caffeine, caused a transient increase in [Ca2+]i and contraction in the absence of external Ca2+ (EGTA, 1 mM). 5-HT inhibited both the carbachol-induced increases in [Ca2+]i release and contractile force. In the beta-escin permeabilized myometrium, 5-HT significantly inhibited the Ca(2+)-induced contraction. The present results indicate that 5-HT stimulates tissue cyclic AMP production, and inhibits the porcine uterine muscle contractility by a reduction in [Ca2+]i and in Ca2+ sensitivity of the contractile elements. Activation of K+ channels might be partially involved in 5-HT-induced inhibition of the myometrial contractility. PMID- 10466939 TI - Evaluation of an agonist index: affinity ratio for compounds active on muscarinic cholinergic M2 receptors. AB - A protocol for predicting full agonist, partial agonist, and antagonist profiles of compounds with M2 muscarinic cholinergic receptor activity was developed using radioligand binding assay techniques with [3H]-N-methyl scopolamine (NMS) and [3H]-Oxotremorine-M (Oxo-M) as radioligands. Full muscarinic cholinergic receptor agonists such as muscarine and oxotremorine-M expressed a high agonist index (> 3000 for M1 muscarinic cholinergic receptors and > 900 for M2 muscarinic cholinergic receptor), whereas muscarinic receptor antagonists (selective or non selective) for different receptor subtypes gave a low (0.5-10) agonist index. Functional studies performed on preparations of guinea-pig ileum and heart were consistent with radioligand binding assay experiments. The above results suggest that similarly as already established for the M1 muscarinic cholinergic receptor subtype, evaluation of the [3H]-NMS/[3H]-Oxo-M ratio may provide useful information on the profile of compounds acting at the M2 muscarinic cholinergic receptor subtype. The availability of simple and predictive techniques for the characterization of muscarinic M2 cholinergic receptor agonists, may help the identification of new compounds in therapeutic areas in which stimulation or inhibition of this receptor is desirable. PMID- 10466940 TI - Evidence for involvement of nitric oxide (NO) or a related nitroso-compound in NANC inhibitory neurotransmission in the pigeon oesophageal smooth muscle. AB - In the pigeon oesophageal smooth muscle electrical field stimulation (EFS), in the presence of atropine and guanethidine, evoked TTX-sensitive inhibitory effects on both the electrical and mechanical activity. N(omega)-Nitro L-arginine (L-NA) (0.1-100 microM), an inhibitor of nitric oxide synthase, reduced the inhibitory EFS-evoked effects. Sodium nitroprusside (SNP) (10 microM), a NO donor, mimicked the effects evoked by EFS. Apamin (1 microM) perfusion did not modify the inhibitory effects induced by SNP. Cystamine (10 microM), a guanylate cyclase inhibitor, reduced the inhibitory effects elicited by EFS. This study shows a possible role for NO in the non-adrenergic non-cholinergic (NANC) inhibitory responses induced by EFS in the pigeon oesophagus. PMID- 10466941 TI - Characterisation of the atypical beta-adrenoceptor in rabbit isolated jejunum using BRL 37344, cyanopindolol and SR 59230A. AB - The present study was carried out to further investigate the nature of the beta adrenoceptor in rabbit jejunum using BRL 37344, a selective beta3-adrenoceptor agonist, cyanopindolol, a beta-adrenoceptor antagonist with blocking activity at beta3-adrenoceptors and SR 59230A, a new selective beta3-adrenoceptor antagonist. Isoprenaline produced a concentration-dependent inhibition of the spontaneous contractions of rabbit jejunum with a pD2 of 7.14. Propranolol (1 microM) shifted the isoprenaline concentration-response curve (CRC) to the right with a concentration-ratio of 5.85, considerably less than would be expected for an action at classical beta-adrenoceptors (estimated pA2 6.66). BRL 37344 also produced a concentration-dependent inhibition of spontaneous contractions with a pD2 of 7.41. The BRL 37344 CRC was unaffected by propranolol (1 microM). In the presence of propranolol (1 microM), cyanopindolol (1 microM) shifted the isoprenaline CRC to the right (concentration-ratio of 21). Cyanopindolol also shifted the BRL 37344 CRC to the right (concentration-ratio of 38). These shifts are consistent with the affinity of cyanopindolol for beta3-adrenoceptors (estimated pA2 values of 7.27 and 7.38 against isoprenaline and BRL 37344, respectively). In the presence of propranolol (1 microM), SR 59230A produced a concentration-dependent rightward shift of the isoprenaline CRC. The Schild plot gave a pA2 value of 7.16, although the slope of the regression line was significantly different from unity (0.65). SR 59230A also produced a concentration-dependent shift of the BRL 37344 CRC. The Schild plot gave a pA2 of 7.58 with the slope of the regression line not significantly different from unity (0.81). The presence of beta3-adrenoceptors mediating relaxation of spontaneous contractions in rabbit jejunum is supported by the relatively poor antagonism of isoprenaline by propranolol, the relaxant effect of BRL 37344 and the antagonism of isoprenaline and BRL 37344 by cyanopindolol and SR 59230A. The lack of simple competitive antagonism of isoprenaline, but not BRL 37344, by SR 59230A may suggest more than one population of atypical beta-adrenoceptor. PMID- 10466942 TI - Effect of early stage of experimental diabetes on vascular functions in isolated perfused kidneys. AB - The present study investigates the renal vascular responsiveness to vasoactive agents in diabetic rats which present an early stage of renal failure. Adult male Wistar rats were administered alloxan (150 mg kg(-1), s.c.). Seven days later the right kidneys were isolated and perfused. Renal perfusion pressure was measured continuously. Concentration-response curves were plotted for noradrenaline (NA), sodium nitroprusside (SNP) and carbachol. In basal conditions, kidneys from diabetic rats presented a decreased vascular resistance compared with those from control rats. The vasoconstrictor response to NA showed decreased EC50 values in preparations from diabetic rats compared with control ones (EC50 nmols, control: 2.03 +/- 0.44, n = 8; diabetic: 0.84+/-0.18, n = 6, P < 0.05). This enhanced sensitivity to NA could be in line with the decreased glomerular filtration rate and cortical renal plasma flow previously described in vivo in our laboratory (Garcia, Girardi, Ochoa, Torres & Elias, 1998). Vasoconstrictor responses to phenylephrine were not however, different between diabetic and control rat kidneys. This suggests that the increased sensitivity to NA was due to impaired neuronal uptake since phenylephrine is not a substrate for neuronal uptake. After precontraction with phenylephrine, both endothelium-dependent (carbachol) and endothelium independent (SNP) vasodilator agents caused similar response in the preparations taken from the two groups of animals. So, the enhanced sensitivity to NA is not associated with a deficient dilator responsiveness of the renal vasculature. The vasodilator response to carbachol was the same in absence or presence of L-arginine in the perfusate, suggesting no alteration in its availability at this stage of diabetes. Diabetic animals showed increased plasma level of fructosamine and glycosylated haemoglobin (Hb A1c), indicating the presence of early glycated products at this stage of diabetes, which could be involved in a possible structural alteration of the vessels. PMID- 10466943 TI - Acute and persistant effects of smoking on the baroreceptor function. AB - Recent studies showed that to smoke four cigarettes within one hour impairs baroreflex sensitivity in humans. In the present study, these effects were qualified more precisely from blood pressure and heart rate records by a sequence analysis and by Fourier analysis of Finapres-registrations. The Mayer waves of the heart rate PDS (power density spectrum), partially representing sympathetic activity, increased during smoking (83.7 +/- 1.0 AU to 89.5 +/- 1.1 AU, P < or = 0.05) and decreased after smoking (86 +/- 1.0 AU, P < or = 0.05). They did, however, not reach baseline levels again within 30 min. Probably due to this, mean arterial blood pressure (64.3 +/- 1.3 mmHg vs. 76.9 +/- 1.3 mmHg, P < 0.05) and heart rate (71.8 +/- 1.4 min(-1) vs. 82.9 +/- 1.4 min(-1), P < 0.05) increased unequivocally after smoking. On the other hand, baroreflex sensitivity decreased dramatically from 15.4 +/- 1 to 11.2 +/- 0.6 ms mmHg(-1) (P < 0.05). This finding was associated with an increased heart rate variability after smoking (6 +/- 0.5 min(-1) vs. 9.2 +/- 1 min(-1)) Thus, the present study provides evidence that chronic tobacco (nicotine)-abuse causes pathologic alterations of the baroreflex control. In synergism with other processes like elevated catecholamine blood levels, these alterations may contribute to the higher risk of cardiovascular diseases. PMID- 10466944 TI - Effect of venlafaxine hydrochloride in different preparations of isolated guinea pig and rat organ tissues. AB - A study was undertaken to know better the effects of venlafaxine hydrochloride on the responses of isolated rat vas deferens to noradrenaline and dopamine, those of isolated rat uterus to serotonin and histamine, and those of isolated guinea pig ileum to acetylcholine and histamine. Venlafaxine hydrochloride increased the response of rat vas deferens to noradrenaline but not to dopamine. Venlafaxine did not alter the response of rat isolated uterus to serotonin. In rat uterus, venlafaxine did not modify the response to histamine but was able to increase it in guinea-pig ileum. An anticholinergic effect was observed with the lowest concentration tested. Although venlafaxine is a selective serotonine reuptake inhibitor in the central nervous system, serotonin uptake was not seen in the rat uterus. The anticholinergic effects observed in the present study might be consistent with some of the side-effects associated with venlafaxine. PMID- 10466945 TI - Myosin light chain kinase inhibitors and calmodulin antagonist inhibit Ca(2+)- and ATP-dependent catecholamine secretion from bovine adrenal chromaffin cells. AB - We have used stage-specific assays for ATP-dependent priming and for Ca(2+) activated triggering in the absence of ATP to examine the effects of myosin light chain kinase (MLCK) inhibitors, ML-9 and ML-7, and calmodulin antagonists, W-7 and trifluoperazine (TFP), on regulated exocytosis in beta-escin-permeabilized bovine adrenal chromaffin cells. Ca2+ (0.1-30 microM) induced a significantly greater secretion of catecholamines in the presence of MgATP (2 mM) than in the absence of MgATP. ML-9 (30 and 100 microM), ML-7 (30 and 100 microM), W-7 (30 and 100 microM) and TFP (10 and 30 microM) inhibited the Ca(2+)-induced catecholamine secretion in the presence of MgATP, but did not affect the catecholamine response to Ca2+ in the absence of MgATP. In intact cells all these compounds inhibited catecholamine secretion in responses to acetylcholine (100 microM) and high K+ (40 mM). The results obtained in permeabilized cells suggest that the calmodulin MLCK system plays an essential role in the ATP-requiring priming stage but not in the Ca2(+)-triggered fusion step in the exocytotic process in bovine adrenal chromaffin cells. PMID- 10466947 TI - University of Bologna and medical knowledge. PMID- 10466946 TI - Changes of cardiac calcium homeostasis in spontaneously hypertensive rats. AB - The aim of the present study was to assess the alterations in cardiac Ca2+ homeostasis induced by hypertension using electrically paced right ventricular strips from Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). 2 Basal contractile force was higher in SHR than in WKY. Similarly, the beta-adrenoceptor agonist isoprenaline (10 nM-10 microM) induced a concentration-dependent positive inotropic effect that was higher in SHR than in WKY, which was in turn inhibited by the beta-adrenoceptor antagonist propranolol (1 microM) in both strains. 3 Preincubation of strips with the L-type Ca2+ channel blockers, nifedipine (1 microM) or verapamil (10 microM), markedly inhibited the isoprenaline response, the inhibition being higher in SHR than in WKY. However, this inhibition was minor by the T-type Ca2+ channel blocker mibefradil (10 microM). 4 Bay K 8644 (10 nM-10 microM), a L-type Ca2+ channel activator induced a concentration-dependent positive inotropic effect, that was greater in SHR than WKY. 5 Nifedipine and verapamil (both 0.1 nM-10 microM) inhibited in a concentration-dependent way the inotropic effect induced by 0.3 microM isoprenaline or 1 microM Bay K 8644. The inhibition was higher in SHR than in WKY. Mibefradil (0.1 nM-10 microM) only clearly inhibited the isoprenaline and Bay K 8644 inotropic effects at 10 microM in both strains. 6 The inhibitor of the sarcoplasmic reticulum Ca2+ release, ryanodine (10 nM-10 microM), was a more effective depressor of isoprenaline induced response in SHR than in WKY. 7 These results suggest that cardiac Ca2+ homeostasis in SHR ventricular strips is altered compared with those of WKY, showing an increased Ca2+ entry through L-type Ca2+ channels and release from sarcoplasmic reticulum; the participation of T-type Ca2+ channels are irrelevant in this tissue. PMID- 10466948 TI - Carbonyl stress: increased carbonyl modification of proteins by autoxidation products of carbohydrates and lipids in uremia. PMID- 10466949 TI - Do shorter hemodialyses increase the risk of death? PMID- 10466950 TI - Correction of CAPD catheter displacement using gastric biopsy forceps: the push pull method. AB - We corrected malpositioned continuous ambulatory peritoneal dialysis catheters in six patients using a new technique named the "push-pull method". A gastric biopsy forceps was advanced through the catheter to near its tip. After manipulating the tip of the forceps through the abdominal wall, the forceps was opened and pulled out slowly. Repeated insertion and removal of the forceps induced the catheter to return to the pelvic cavity. This push-pull method was successful for Swan neck straight (n=2) and coiled (n=4) catheters in all patients. The time required for the procedure was only 5-10 minutes and there were no complications. PMID- 10466951 TI - On-line monitoring of the intravascular volume during haemodialysis by continuous refractometry. AB - The control of intravascular volume (IVV) by continuous on-line measurement of protein concentration would optimise the patients' specific rate of ultrafiltration. To prove the accuracy of a refractometric device, plasma was continuously drawn by haemofiltration during 10 haemodialysis treatments of male patients. Refractometry reflects highly significant changes in the concentrations of filtrate proteins (r = 0.862, p < 0.001) and blood proteins (rtotal = 0.593, ptotal < 0.001). In vitro, the refractometric device detected a change of protein concentration of 0.041 g/L through a refraction increase of 0.1 mV. The power of discrimination was 0.067% of IVV However, in vivo, the accuracy of IVV refractometric monitoring is reduced by interference factors such as sodium (0. 141 mV/mmol/L), glucose (0.034 mV/mg/dl) and triglycerides (-0.040 mV/mg/dl). Adjustment of the refraction data using sodium and glucose electrodes and plasma filters with a cut-off below the size of chylomicrons is recommended. PMID- 10466952 TI - Effects of unprocessed and processed cardiopulmonary bypass blood retransfused into patients after cardiac surgery. AB - BACKGROUND: The aim of this prospective study was to compare the effect of autologous unprocessed to processed residual cardiopulmonary bypass blood (CPB) on patients' laboratory and clinical parameters and outcome. METHODS: 20 patients undergoing elective coronary artery bypass surgery were randomized to receive either unprocessed CPB blood (control group) or processed CPB blood employing the Continuous AutoTransfusion System (CATS; Fresenius, Bad Homburg, Germany). We have shown that this method eliminated >93% of activated mediators. Serial laboratory parameters including complement activation, coagulation factors and the stimulation of IL-6 and IL-8 were compared with clinical side effects and patients' outcome. RESULTS: Compared to control patients, retransfusion of unprocessed CBP blood significantly increased heparin, free plasma hemoglobin and D-Dimers. Postoperatively, three patients in the control group and two patients in the CATS group required prolonged mechanical ventilation or developed infections associated respectively with elevated C3a (desArg) or IL-6 concentration. CONCLUSIONS: CATS-processing of CPB blood provided a high-quality red blood cell concentrate, resulting in a reduced load of retransfused activated mediators. PMID- 10466953 TI - A simple method for Emax trend evaluation: in vitro and in vivo results. AB - The aim of this study is the evaluation of end systolic ventricular elastance trend (as a measure of heart contractility) by hemodynamic variables available in intensive care units or during heart surgery: heart rate, cardiac output, left atrial, mean and diastolic arterial pressure. Its basic assumption is the description of ejection as the interaction between variable left ventricular and arterial compliances (reciprocal of the corresponding elastances) connected in parallel. As pressure is the same in each compliance at systole beginning and ending, ventricular elastance can be estimated by assuming that energy variation is the same on both compliances. The algorithm has been tested on a numerical simulator of the circulatory system and on six sheep at basal conditions and during drug infusion. Correlation function in numerical simulation, between true and computed ventricular elastance (range 0.45 divided by 5 mm Hg-cm(-3)), yields 0.985. In vivo comparison between computed ventricular elastance trend and ventricular dp/dt trend yields a correlation function ranging between 0.87 and 0.99. The result of the algorithm cannot be assumed to be Emax value. However, it can be considered a contractility index as it closely follows any change in dp/dt. It can be computed by simple calculations and needs no variables other than those usually measured in intensive care . It allows the extrapolation of useful information for evaluating the trend in heart contractility and for setting up a control strategy for mechanical or pharmacological assistance during heart recovery. PMID- 10466954 TI - Hip prothesis: an in vitro wear protocol based on a comparison between gravimetric and profilometric analysis. AB - Ultra high molecular weight polyethylene acetabular cups were analysed by means of a shadowgraph method (using a profile projector) to measure linear wear. The results were compared with those of previous wear tests performed on a hip joint simulator. Twelve polyethylene acetabular cups were analysed. The specimens were evaluated visually for evidence of polyethylene wear. Examination of the polyethylene inner surface did not reveal evidence of surface failure such as delamination, fatigue cracks or scratches. Volumetric wear was calculated using a formula based on dimensional change due to the penetration of the femoral head in the acetabular cup. It was found to be of the same order of magnitude as the wear obtained in in vitro experimental tests. PMID- 10466955 TI - Autotransfusion with predeposit-haemodilution and perioperative blood salvage: 20 years of experience. Rizzoli Study Group on Orthopaedic Anesthesia. AB - BACKGROUND: Through a prospective study, we evaluated the efficacy of an integrated autotransfusion regimen comprised of predeposit-hemodiluition and intra and post-operative blood salvage in major orthopaedic surgery. METHODS: We examined the records of 2303 consecutive patients (1524 females and 779 males, mean age 62.7, standard deviation 11 years (range 16-90 yrs), pre-operative haemoglobin (Hb) concentration 13.5 (SD 1.4) (range 6.7-19.3) g/dl undergong total hip arthroplasty (THA 1582 patients), THA after the removal of internal fixation devices (RFD+THA, 25 patients), total knee arthroplasty (TKA, 347 patients), revision surgery of the hip (HR, cup+stem revision, 248 patients; cup revision 64 patients; stem revision 23 patients) and total knee revision (TKR 14 patients). We estimated that the number of predonations (MSBOS - maximum surgery blood order schedule) was 2 units for THA, TKA and TKR, and 3 units for partial or total hip revision and for total hip arthroplasty with fixation removal. RESULTS: It was possible to obtain the MSBOS in 2070 patients (89.8%). Homologous red blood cell (HRBC) transfusion were carried out in 184 patients (8%). We found that the need to use HRBC was significantly associated with failure to meet the number of MSBOS, female sex, lower pre-operative Hb concentration, use of calcium heparin for antithrombosis prophylaxis, more extensive surgery, a higher ASA rating and co-existing diseases such as coronary artery disease. CONCLUSIONS: Cooperation among anaesthesis, transfusionists and surgeons in the application of an integrated autotransfusion regimen enabled us to treat 92% of our patients with only autotransfusion. PMID- 10466956 TI - Functionally adapted surfaces on a silicone keratoprosthesis. AB - BACKGROUND: Silicone intraocular lenses as well as silicone sponges and encircling bands on the bulbar surface are widely used and are well tolerated. The aim of this project is a new one-piece silicone keratoprosthesis with enhanced cell adhesion in the haptic region to optimize the keratoprosthesis stability. These investigations show how enhanced profileration of conjunctival fibroblasts and, therefore, improved tissue compatibility can be achieved by hydrophilizing and by protein immobilisation on a hydrophobic silicone surface. This allows a combination of desired chemical and mechanical properties of the silicone bulk material with surfaces of improved tissue compatibility. METHODS: Silicone foils with surface modifications of different kinds were tested. Experiments were done using cell cultures with murine fibroblasts L-929 and human conjuctival fibroblasts. Cytotoxicity assays were carried out with cells grown on the material in direct contact, as well as in indirect contact, with extracts (EN 30993-5). Viability stains by means of fluoresceindiacetate and ethidiumbromide together with morphology analyses by hemalaun-staining were performed. RESULTS: For the unmodified and modified foils themselves and their extracts any negative influence on cell cultures of murine and human cells could be excluded. There was a gradual improvement of cell morphology, spreading and proliferation dependent on the degree of surface modification. Covalently immobilised fibronectin showed the best results in contrast to adsorptive binding. CONCLUSIONS: Silicone surfaces can be modified chemically with bioactive proteins. These modifications are cell compatible and do not result in toxic reactions. The degree and type of silicone hydrophilization results in improved development of cell morphology, spreading and proliferation. Even better results are obtained after covalent binding of bioactive proteins like fibronectin. Improved biocompatibility with enhanced cellular overgrowth has been demonstrated in vitro for the modified silicone of the haptic region. We believe that this type of modification will help in reducing extrusion problems observed with former keratoprostheses. PMID- 10466957 TI - An intranasal challenge model for testing Japanese encephalitis vaccines in rhesus monkeys. AB - Placebo-controlled field efficacy trials of new Japanese encephalitis (JE) vaccines may be impractical. Therefore, an animal model to evaluate efficacy of candidate JE vaccines is sought. Previous work has shown that exposure of monkeys to JE virus (JEV) via the intranasal route results in encephalitis. Here we report the further development of this model and the availability of titered virus stocks to assess the protective efficacy of JE vaccines. To determine the effective dose of our JE challenge virus, dilutions of a stock JEV (KE-93 isolate) were inoculated into four groups of three rhesus monkeys. A dose dependent response was observed and the 50% effective dose (ED50) was determined to be 6.0 x 10(7) plaque forming units (pfu). Among animals that developed encephalitis, clinical signs occurred 9-14 days postinoculation. Infection with JEV was confirmed by detection of JEV in nervous tissues and IgM to JEV in the cerebrospinal fluid. Viremia with JEV was also detected intermittently throughout infection. Validation of the model was performed using a known effective JE vaccine and saline control. One ED90 of virus (2.0 x 10(9) pfu) was used as a challenge dose. Four of four animals that received saline control developed encephalitis while one of four monkeys administered the JE vaccine did so. This study demonstrates that the virus strain, route of inoculation, dose, and the outcome measure (encephalitis) are suitable for assessment of protective efficacy of candidate JE vaccines. PMID- 10466958 TI - Production of lethal infection that resembles fatal human disease by intranasal inoculation of macaques with Japanese encephalitis virus. AB - Twelve rhesus macaques (Macaca mulatta) challenged intranasally with a wild-type Japanese encephalitis virus (JEV) developed clinical signs 11-14 days later. Tissues from the cerebral cortex, cerebellum, brainstem, thalamus, meninges, and all levels of the spinal cord were stained for JEV antigen with hyperimmune mouse ascitic fluid and streptavidin-alkaline phosphatase; immunofluorescent staining was also done on frozen sections. Viral antigen was found in all cell layers of the cerebellum, the gray matter of the thalamus and brainstem, and the ventral horn of all levels of the spinal cord. Staining was limited to neurons and their processes. Histopathologic changes were limited to the nervous system and characterized by nonsuppurative meningoencephalitis. These results were comparable with those of previous studies done with human autopsy tissues. Intranasal inoculation of rhesus monkeys with JEV was effective in producing clinical disease comparable with natural disease in humans and may serve as a model to evaluate protective efficacy of candidate JEV vaccines. PMID- 10466959 TI - Safety, immunogenicity, and protective efficacy of NYVAC-JEV and ALVAC-JEV recombinant Japanese encephalitis vaccines in rhesus monkeys. AB - Two poxvirus-vectored vaccines for Japanese encephalitis (JE), NYVAC-JEV and ALVAC-JEV, were evaluated in rhesus monkeys for safety, immunogenicity, and protective efficacy. The vaccines were given to four monkeys each on study days 0 and 28 along with saline placebo on day 7. For controls, the licensed BIKEN JE vaccine and a saline placebo were given to other groups of four monkeys on days 0, 7, and 28. No systemic effects were observed. All injection site reactions were mild. All vaccines elicited appreciable JE-specific neutralizing antibody responses. However, a more rapid increase and higher peak level of antibody were seen in the BIKEN group as compared with the NYVAC-JEV and ALVAC-JEV groups. The peak neutralizing antibody level in the NYVAC-JEV group was higher than that of the ALVAC-JEV group. Antibody persisted in all four BIKEN recipients through 273 days of follow-up, whereas, the antibody level decreased to the threshold of detection in two NYVAC-JEV and all four ALVAC-JEV recipients by day 120. On day 273, all monkeys were given a booster dose. A rapid increase in neutralizing antibody was seen in all vaccine recipients by seven days. Two months after the booster dose, all monkeys were challenged intranasally with one 90% effective dose of JE virus. Four recipients of saline, three of ALVAC-JEV, one of NYVAC JEV, and one of BIKEN experienced encephalitis. This study suggests that the NYVAC-JEV and ALVAC-JEV vaccines are safe and immunogenic in monkeys and that the NYVAC-JEV and BIKEN vaccines are effective in protecting monkeys from encephalitis. PMID- 10466960 TI - Testing the efficacy of a recombinant merozoite surface protein (MSP-1(19) of Plasmodium vivax in Saimiri boliviensis monkeys. AB - Saimiri boliviensis monkeys were immunized with the yeast-expressed recombinant protein yP2P30Pv200(19). The antigen consisted of the C-terminus (amino acid Asn1622-Ser1729) of the merozoite surface protein 1 of the Plasmodium vivax Salvador I strain. Two universal T helper cell epitopes (P2 and P30) of tetanus toxin and six histidine residues for purification purposes were attached to the N and C-termini, respectively. Four groups of five monkeys were given three immunizations at four-week intervals with either 250 microg of yP2P30Pv200(19) formulated with nonionic block copolymer P1005, 250 microg of antigen adsorbed to alum, 250 microg of antigen in phosphate-buffered saline (PBS), or PBS alone. Five weeks after the last immunization, each animal was inoculated with 100,000 parasitized erythrocytes of the Salvador I strain of P. vivax. Animals were splenectomized one week after challenge to increase parasite densities; after seven weeks of infection, animals were treated. Eighteen weeks later, the animals were rechallenged with the homologous parasite. Following the first challenge, three monkeys immunized with the antigen with P1005 were protected; no animals were protected from rechallenge. One monkey immunized with yP2P30Pv200(19) with alum was protected; no protection was seen after rechallenge. Two monkeys immunized with antigen alone were protected; none were protected from rechallenge. One control animal had a low parasite count following primary infection; none were protected against rechallenge. Adverse reactions were only observed with animals receiving P1005. It is proposed that splenectomy of the monkeys prevented adequate assessment of the efficacy of this antigen. Identification of a monkey host that supports high density parasitemia without splenectomy appears needed before further testing of blood-stage vaccines against P. vivax. PMID- 10466961 TI - Longevity of naturally acquired antibody responses to the N- and C-terminal regions of Plasmodium vivax merozoite surface protein 1. AB - In an earlier study, we found that individuals with patent infection had significantly higher IgG antibody titers to the 19-kD C-terminal region of Plasmodium vivax merozoite surface protein 1 (PvMSP1) than individuals treated for malaria 1-4 months earlier. These results suggested that the antibody levels decreased rapidly following treatment. The present study was designed to determine the persistence of antibody response to the N- and C-terminal regions of PvMSP1 after infection with P. vivax in individuals from the city of Belem in northern Brazil. Our results demonstrated that the vast majority of individuals had a significant decrease in antibody titers to the C-terminal region of PvMSP1 in a period of two months following treatment. Among responders to the C-terminal region, 44.4% became serologically negative and 44.4% had their antibody titers reduced by an average of 13-fold. Only 11.2% of the individuals had their antibody titers maintained or slightly increased during that period. A decrease in the antibody response to the recombinant protein representing the N-terminal region of PvMSP1 was also noted; however, it was not as dramatic. The rapid decrease in the antibody levels to the C-terminal region of PvMSP1 might contribute to the high risk of reinfection in these individuals. PMID- 10466962 TI - Population structure of the primary malaria vector in South America, Anopheles darlingi, using isozyme, random amplified polymorphic DNA, internal transcribed spacer 2, and morphologic markers. AB - A genetic and morphologic survey of Anopheles darlingi populations collected from seven countries in Central and South America was performed to clarify the taxonomic status of this major malaria vector species in the Americas. Population genetics was based on three techniques including isozyme, random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), and internal transcribed spacer 2 (ITS2) markers. The results of the isozyme analysis indicated moderate differences in the allele frequencies of three putative loci (glutamate oxalaoacetate transaminase-1, isocitrate dehydrogenase-1, and phosphoglucomutase) of the 31 analyzed. No fixed electromorphic differences separated the populations of An. darlingi, which showed little genetic divergence (Nei distances = 0.976 0.995). Fragments produced by RAPD-PCR demonstrated evidence of geographic partitioning and showed that all populations were separated by small genetic distances as measured with the 1 - S distance matrix. The ITS2 sequences for all samples were identical except for four individuals from Belize that differed by a three-base deletion (CCC). The morphologic study demonstrated that the Euclidean distances ranged from 0.02 to 0.14, with the highest value observed between populations from Belize and Bolivia. Based on these analyses, all the An. darlingi populations examined demonstrated a genetic similarity that is consistent with the existence of a single species and suggest that gene flow is occurring throughout the species' geographic range. PMID- 10466963 TI - Mitochondrial DNA sequence variation among triatomine vectors of Chagas' disease. AB - Kissing bugs or triatomines (Reduviidae: Triatominae) are vectors of the Chagas' disease agent Trypanosoma cruzi. There is a current need for more sensitive tools for use in discrimination of different bug populations and species, thus allowing a better understanding of these insects as it relates to disease transmission and control. In a preliminary analysis of the mitochondrial large subunit ribosomal RNA (mtlsurRNA) and cytochrome B (mtCytB) genes, we used DNA sequencing to study species identification and phylogeny. In both examined gene regions, about 46% of nucleotide positions exhibited polymorphism. The examined region of mtCytB appears to have evolved more rapidly than the examined region of mtlsurRNA. Phylogenetic analysis of both gene fragments in the examined species produced similar results that were generally consistent with the accepted taxonomy of the subfamily. The two major tribes, Rhodniini and Triatomini, were supported, along with additional clades that corresponded to accepted species complexes within the Rhodnius and Triatoma genera. The one chief exception was that Psammolestes coreodes sorted into the Rhodnius prolixus-robustus-neglectus clade, with bootsrap values of 99% and 81%, respectively, for the mtlsurRNA and mtCytB fragments. All of the individual species examined could be distinguished at both genetic loci. PMID- 10466964 TI - Eastern equine encephalitis virus in birds: relative competence of European starlings (Sturnus vulgaris). AB - To determine whether eastern equine encephalitis (EEE) virus infection in starlings may be more fulminant than in various native candidate reservoir birds, we compared their respective intensities and durations of viremia. Viremias are more intense and longer lasting in starlings than in robins and other birds. Starlings frequently die as their viremia begins to wane; other birds generally survive. Various Aedes as well as Culiseta melanura mosquitoes can acquire EEE viral infection from infected starlings under laboratory conditions. The reservoir competence of a bird is described as the product of infectiousness (proportion of feeding mosquitoes that become infected) and the duration of infectious viremia. Although starlings are not originally native where EEE is enzootic, a starling can infect about three times as many mosquitoes as can a robin. PMID- 10466965 TI - The effect of iron on the toxigenicity of Vibrio cholerae. AB - In vitro and in vivo studies were conducted to assess the response of cholera toxin (CT) production to increasing iron concentrations in an aquatic environment. Production of CT by seven of eight Vibrio cholerae strains tested, including the Bengal strain (O139), was significantly enhanced in the presence of iron concentrations of 1.0 and 10 g/L. The exception (El Tor Ogawa) had a significant CT response only in the presence of 10 g of iron/L. Enhancement of CT production also occurred at iron concentrations less than 1.0 g/L, but not to a statistically significant degree. The high iron concentrations, which in this study were found to stimulate CT production, have been described by others in association with sediments, water plants, and chitinous fauna. Other investigators have shown a predilection by V. cholerae to attach to these sites in the aquatic environment. The importance of excess in vivo iron with respect to the pathogenicity of several gram-negative bacilli is well recognized. However, the possible impact of environmental iron on the in vitro toxigenicity of a microorganism, in this case V. cholerae in its aquatic environment, is to the best of our knowledge a new finding with important epidemiologic implications. These findings, coupled with the fact that iron concentration is considerably enhanced in industrially polluted waters and sediments, may reflect a causal link between the concurrent global upsurge of industrialization and pandemic occurrence of cholera during the latter half of the 20th century. Enhanced toxigenicity may also cause clinical disease following ingestion of lower than usual infective doses of cholera vibrios, thereby increasing the incidence of symptomatic cases and, possibly, of severe cases. PMID- 10466966 TI - Reliability of parental history of antibiotic use for Filipino children admitted with acute lower respiratory tract infection. AB - Parental history on antibiotic use and the urine antibacterial assay (UABA) result were compared in a study on Filipino children with acute lower respiratory tract infection (ALRI). Among 108 patients in whom urine for the UABA could be collected prior to starting antibiotic treatment in the hospital, 59 (55%) guardians reported preceding antibiotic use, 54% of whom were positive in the UABA. In another 37 (34%), the UABA result was positive, indicating nonreported use of antibiotics. Among 190 patients in whom urine could be collected only after intravenous administration of antibiotic, the UABA demonstrated large inhibition zones after the first dose in most patients but a negative result was seen in 14 cases. The inhibition zone radius was significantly smaller for chloramphenicol than for beta-lactam antibiotics (8.3 mm versus 16.1 mm after one dose; 95% confidence intervals = 7.0-9.7 and 14.9-17.2, respectively). Parental history on antibiotic use gives an underestimate of preceding antibiotic use in children with ALRI in the Philippines. The result partly explains the low yield of blood culture in many studies on ALRI, and stresses the need to develop new diagnostic methods not based on culture for those organisms highly sensitive to antibiotics such as Streptococcus pneumoniae and Haemophilus influenzae. PMID- 10466967 TI - Intestinal blockage by carcinoma and Blastocystis hominis infection. AB - We detected heavy infections of Blastocystis hominis in four individuals with intestinal obstruction due to cancerous growths. After surgery, the infections spontaneously resolved, without specific chemotherapy. It appears that the B. hominis infection was coincidental and not related to the neoplastic growth. We suggest that intestinal obstruction and concomitant stool retention, plus hemorrhage from cancerous lesions, may have permitted the more abundant growth of B. hominis. This is the first report of a possible relationship between intestinal obstruction and a concomitant B. hominis infection. PMID- 10466968 TI - Recovery of a second instar Gasterophilus larva in a human infant: a case report. AB - We report a case in an infant of horse bot fly myiasis that was unusual because the maggot had developed to the second instar (of three potential instars). This represents the first report of such late development in a human. The case occurred in a rural area of the Pacific northwest (Washington) in late summer. PMID- 10466969 TI - Ocular linguatuliasis in Ecuador: case report and morphometric study of the larva of Linguatula serrata. AB - Linguatula serrata is a pentastomid, a cosmopolitan parasite belonging to the Phylum Pentastomida. Humans may act as an intermediate or accidental definitive host of this parasite, manifesting the nasopharyngeal or visceral form, with the latter having been described more frequently. The occurrence of ocular linguatuliasis is extremely rare, but it has been reported in the United States and Israel. The objective of the present paper was to report the first case of ocular linguatuliasis in Ecuador and to extend the morphologic study of L. serrata by morphometric analysis. The patient studied was a 34-year old woman from Guayaquil, Ecuador who complained of ocular pain with conjunctivitis and visual difficulties of two-months duration. Biomicroscopic examination revealed a mobile body in the anterior chamber of the eye. The mobile body was surgically removed. The specimen was fixed in alcohol, cleared using the technique of Loos, stained with acetic carmine, and mounted on balsam between a slide and a coverslip. It was observed with stereoscopic and common light microscopes in combination with an automatic system for image analysis and processing. The morphologic and morphometric characteristics corresponded to the third-instar larval form of L. serrata. To our knowledge, ocular linguatuliasis has not been previously described in South America, with this being the first report for Ecuador and South America. The present study shows that computer morphometry can adequately contribute both to the morphologic study and to the systematic classification of Pentastomids, and L. serrata in particular. PMID- 10466970 TI - Plasmodium falciparum clinical malaria in Dielmo, a holoendemic area in Senegal: no influence of acquired immunity on initial symptomatology and severity of malaria attacks. AB - Six hundred eighty-nine Plasmodium falciparum malaria attacks were observed during a three-year period among 226 inhabitants of the village of Dielmo, Senegal, an area of high malaria transmission. Malaria attacks were defined as clinical episodes with fever (body temperature > or = 38.0 degrees C) or reporting of fever or headache or vomiting, associated with a parasite:leukocyte ratio above an age-dependent pyrogenic threshold identified in this population. The symptom frequencies were tested against age, gender, and parasite density using a random-effect logistic regression model and the study of distinguishable clinical presentations was carried out by multi-correspondence analysis. There was little difference between the severity of symptoms during the initial course of attacks in young children and adults, and this severity was not correlated with the duration of the pathologic episode. It was not possible to distinguish objectively different malaria attack types according to the severity of clinical manifestations. In contrast, the duration of fever, symptoms, and parasite clearance were significantly longer among the youngest children than among the oldest children and adults. These findings suggest that of the two components of protective immunity, anti-parasite immunity and anti-toxic immunity, only the first would play a major role as age increases. They suggest also that the initial clinical presentation of malaria attacks is not predictive of the level of protective immunity. PMID- 10466971 TI - Malaria vectors in a traditional dry zone village in Sri Lanka. AB - Malaria transmission by anopheline mosquitoes was studied in a traditional tank irrigation-based rice-producing village in the malaria-endemic low country dry zone of northcentral Sri Lanka during the period August 1994-February 1997. Adult mosquitoes were collected from human and bovid bait catches, bovid-baited trap huts, indoor catches, and pit traps. Mosquito head-thoraces were tested for the presence of Plasmodium falciparum and P. vivax, and blood-engorged abdomens for the presence of human blood by ELISAs. House surveys were done at two-day intervals to record cases of blood film-confirmed malaria among the villagers. A total of 7,823 female anophelines representing 14 species were collected. Trends in anopheline abundance were significantly correlated with rainfall of the preceding month in An. annularis, An. barbirostris, An. subpictus, An. vagus, and An. varuna, but were not significant in An. culicifacies and An. peditaeniatus. Malaria parasite infections were seen in seven mosquito species, with 75% of the positive mosquitoes containing P. falciparum and 25% P. vivax. Polymorph PV247 was recorded from a vector (i.e., An. varuna) for the first time in Sri Lanka. Computations of mean number of infective vector (MIV) rates using abundance, circumsporozoite (CS) protein rate, and human blood index (HBI) showed the highest rate in An. culicifacies. A malaria outbreak occurred from October 1994 to January 1995 in which 45.5% of village residents experienced at least a single disease episode. Thereafter, malaria incidence remained low. Anopheles culicifacies abundance lagged by one month correlated positively with monthly malaria incidence during the outbreak period, and although this species ranked fifth in terms of abundance, infection was associated with a high MIV rate due to a high CS protein rate and HBI. Abundance trends in other species did not correlate significantly with malaria. It was concluded that An. culicifacies was epidemiologically the most important vector in the study area. PMID- 10466972 TI - Potential for evolution of California serogroup bunyaviruses by genome reassortment in Aedes albopictus. AB - Aedes albopictus was introduced into the United States in used tires in 1985. Its successful colonization of the upper Midwest has potential to alter the current epidemiology of bunyaviruses that circulate in the region. It is permissive for the replication of several arboviruses, including La Crosse (LACV) and Jamestown Canyon (JCV) bunyaviruses. In this study, we demonstrate the ability of LACV and JCV to coinfect Ae. albopictus mosquitoes and to form all six possible reassortant genotypes. All reassortant viruses infect Ae. albopictus orally and can be transmitted to suckling mice. All reassortants are neurovirulent in mice. However, reassortant viruses carrying the LACV M segment in the foreign genetic background of JCV are more neuroinvasive than JCV, or any other reassortant genotype. In addition, these reassortants can replicate in gerbils and infect Ae. triseriatus, characteristics of LACV, but not JCV. Because Ae. albopictus is spreading into new geographic areas and feeds on a variety of mammals, including humans, it has the potential to transmit new, emerging bunyaviruses in nature. PMID- 10466973 TI - Isolation of thogoto virus (Orthomyxoviridae) from the banded mongoose, Mongos mungo (Herpestidae), in Uganda. AB - Small wild vertebrates were trapped during an investigation into possible vertebrate reservoirs of o'nyong-nyong (ONN) fever virus in Uganda in 1997. Antibody neutralization test results and virus isolation attempts were negative for ONN virus, confirming the work of earlier investigators, who also failed to find evidence for a nonhuman ONN virus reservoir. In the course of these ONN virus studies, Thogoto virus was isolated from one of eight banded mongooses (Mongos mungo). This is the first isolation of Thogoto virus from a wild vertebrate. Neutralizing antibodies to Thogoto virus were also found in two of the other mongooses. PMID- 10466975 TI - A continuing focus of Hansen's disease in Texas. AB - To describe epidemiologic and clinical characteristics of Hansen's disease cases in Texas, information was abstracted from records of 810 patients reported from 1973 through 1997. Annually, from 18 to 54 patients were reported. Average annual incidence rates ranged from 1.9 to 2.4 cases per million population. A majority of the patients were male (63%) and white (77%). More than half (53%) of the patients were born in the United States; a majority (83%) of the patients born in the United States were born in Texas. Most (76%) patients were diagnosed with multi-bacillary leprosy. Foreign-born patients were more likely to be younger at onset and have multi-bacillary disease compared with patients born in the United States. Within Texas, an endemic focus of Hansen's disease exists along the Gulf of Mexico coast. PMID- 10466974 TI - Genetic evidence for the origins of Venezuelan equine encephalitis virus subtype IAB outbreaks. AB - Epizootics of Venezuelan equine encephalitis (VEE) involving subtype IAB viruses occurred sporadically in South, Central and North America from 1938 to 1973. Incompletely inactivated vaccines have long been suspected as a source of the later epizootics. We tested this hypothesis by sequencing the PE2 glycoprotein precursor (1,677 nucleotides) or 26S/nonstructural protein 4 (nsP4) genome regions (4,490 nucleotides) for isolates representing most major outbreaks. Two distinct IAB genotypes were identified: 1) 1940s Peruvian strains and 2) 1938 1973 isolates from South, Central, and North America. Nucleotide sequences of these two genotypes differed by 1.1%, while the latter group showed only 0.6% sequence diversity. Early VEE virus IAB strains that were used for inactivated vaccine preparation had sequences identical to those predicted by phylogenetic analyses to be ancestors of the 1960s-1970s outbreaks. These data support the hypothesis of a vaccine origin for many VEE outbreaks. However, continuous, cryptic circulation of IAB viruses cannot be ruled out as a source of epizootic emergence. PMID- 10466976 TI - Isolation of Borrelia burgdorferi from Neotoma fuscipes, Peromyscus maniculatus, Peromyscus boylii, and Ixodes pacificus in Oregon. AB - The number of Lyme disease cases in Oregon has increased in recent years despite the fact that the pathogen, Borrelia burgdorferi, has never been isolated in the state. Rodent and tick surveys were undertaken in 1997 to isolate and characterize strains of B. burgdorferi from Oregon and to identify potential reservoirs and vectors of Lyme disease. Borrelia burgdorferi was isolated from Neotoma fuscipes, Peromyscus maniculatus, P. boylii, and Ixodes pacificus. Both N. fuscipes and P. maniculatus were infested with I. pacificus and I. spinipalpis. Although I. pacificus infested P. boylii, I. spinipalpis was not found on this rodent, and only 4% of the P. boylii were infected with B. burgdorferi compared with the 19% and 18% infection rates found in N. fuscipes and P. maniculatus, respectively. Variation in the molecular weights of the outer surface proteins A and B were found in these first confirmed isolates of B. burgdorferi from Oregon, as well as truncated forms of outer surface protein B. PMID- 10466977 TI - Epidemiology of Burkholderia pseudomallei in Thailand. AB - The distribution of Burkholderia pseudomallei in soil collected from four regions of Thailand and the frequency of B. pseudomallei infections in patients attending government hospitals throughout Thailand in 1997 were surveyed. A total of 3,585 soil samples collected from 896 sites in four regions of Thailand were cultured for B. pseudomallei using selective enrichment broth and modified Ashdown's agar. The organism was recovered in 4.4%, 6.1%, 20.4%, and 5.9% of the soil samples collected from the northern, central, northeastern, and southern regions, respectively, of Thailand (P < 0.0001). Burkholderia pseudomallei was cultured from 50.1% of the sites in the northeastern region compared with 13.8%, 24.5%, and 18.4% in the northern, central, and southern regions, respectively (P < 0.0001). The infection rate in patients attending government hospitals in the northeastern region (137.9 per 100,000 inpatients) was significantly higher than those in the northern (18 per 100,000 inpatients), central (13.4 per 100,000 inpatients), and southern (14.4 per 100,000 inpatients) regions, respectively (P < 0.0001). It is suggested that melioidosis, which is endemic in Thailand, is associated with the presence of B. pseudomallei in soil. PMID- 10466978 TI - An extensive ultrasound and serologic study to investigate the prevalence of human cystic echinococcosis in northern Libya. AB - A prevalence study of abdominal cystic echinococcosis (CE) was undertaken in the northwest, north-central, and northeast regions of Libya. A total of 36 villages along the coast were included, in which 20,220 people were screened by portable ultrasound. Three hundred thirty-nine (1.7%) were diagnosed with CE. There was no significant difference between CE prevalence rates in the three regions of Libya (mean = 1.6%); however, intervillage rates were variable, ranging from 0% to 4.5%. The prevalence of CE increased significantly with age (P < 0.0001) and females were significantly more affected (2%) than males (1.3%) (P < 0.0001). Cases of CE were distributed among 3.2% of housewives, 2.6% of farmers, 2.3% of male civil servants, 1.3% of female students, and 1.1% of male students. Housewives (P < 0.0001) and students (P < 0.0001) were significantly more at risk for CE. A statistically significant proportion (62% [210 of 339]) of CE cases kept dogs (P < 0.0001). Of the ultrasound-positive CE cases 69% (233 of 339) were antibody seropositive by ELISA using Echinococcus granulosus hydatid cyst fluid antigen B. Blood samples (n = 10,096) collected onto filter papers were taken randomly from ultrasound-negative people who entered the study, of which 11.2% were found to be seropositive. Seropositivity increased with age (P < 0.0001), with females exhibiting higher seroprevalence than males for all age groups (P < 0.0001). Seroprevalence was distributed occupationally among 17.3% of female students, 11.2% of male students, 8.3% of housewives, 7.3% of farmers, and 6.4% of male civil servants. Housewives and students were more likely to be seropositive (P < 0.0001). Forty-seven percent (526) of the seroreactors kept dogs. All liver hydatid cysts detected by ultrasound during community screening were classified according to morphology and size into six types. Type I (17.5% of all cases) were small univesicular cysts less than 50 mm in diameter with no laminations or daughter cysts. Type II (34%) were univesicular cysts with only laminations. Type IIIa (8.5%) were univesicular cysts with the appearance of laminations and daughter cysts. Type IIIb (13%) were univesicular with laminations and less prominent daughter cysts. Type IV (5.6%) presented as a solid mass. Type V were degenerated calcified or partially calcified cysts (13.2%). Type VI presented as multiple cysts (8% of all cases). The CE cases that exhibited Types II, IIIa, IIIb, IV or VI cysts showed the highest seropositivity (86%, 96%, 95%, 100%, and 96%, respectively), while Types I and V were the least seroreactive (38% and 22%, respectively). Cases of CE occurred in 311 families, with 93% having only one member as a CE case while 7% of the families had two or more cases. However, 25% of the ultrasound-negative persons belonging to families with an index CE case were seropositive for antibodies to Echinococcus. These results confirm the importance of human CE in Libya. They also confirm the usefulness of ultrasound combined with serology as a mass screening approach for CE in north African communities. PMID- 10466979 TI - Phenotypic and genotypic characteristics of recently adapted isolates of Plasmodium falciparum from Thailand. AB - The drug sensitivity characteristics and Plasmodium falciparum pfmdr1 status of five isolates of P. falciparum recently isolated from patients presenting for treatment from the Thailand/Myanmar border have been investigated. The aim of the study was to avoid the criticisms of some earlier studies by focusing on newly collected isolates from a specific geographic location. Three of the isolates studied exhibited clear resistance to chloroquine similar to that observed in the K1 Thai standard isolate obtained in the 1970s, and the other two isolates were of intermediate sensitivity to chloroquine with concentrations of drug that inhibit parasite growth by 50% of 50 and 43 nmol. The sensitivity of all isolates was enhanced by verapamil but we found no clear association between chloroquine sensitivity and gene copy number or intra-allelic variation of pfmdr1. In contrast, clear cross-resistance was seen between mefloquine and halofantrine, with the most sensitive isolates carrying the K1 mutation in pfmdr1. PMID- 10466980 TI - Pyrimethamine-sulfadoxine efficacy and selection for mutations in Plasmodium falciparum dihydrofolate reductase and dihydropteroate synthase in Mali. AB - To assess pyrimethamine-sulfadoxine (PS) efficacy in Mali, and the role of mutations in Plasmodium falciparum dihydrofolate reductase (DHFR) and dihydropteroate synthase (DHPS) in in vivo PS resistance, 190 patients with uncomplicated P. falciparum malaria were treated with PS and monitored for 56 days. Mutation-specific polymerase chain reactions and digestion with restriction endonucleases were used to detect DHFR and DHPS mutations on filter paper blood samples from pretreatment and post-treatment infections. Only one case each of RI and RII level resistance and no cases of RIII resistance or therapeutic failure were observed. Post-PS treatment infections had significantly higher rates of DHFR mutations at codons 108 and 59. No significant selection for DHPS mutations was seen. Pyrimethamine-sulfadoxine is highly efficacious in Mali, and while the low level of resistance precludes assessing the utility of molecular assays for in vivo PS resistance, rapid selection of DHFR mutations supports their role in PS failure. PMID- 10466981 TI - Assessment of combined ivermectin and albendazole for treatment of intestinal helminth and Wuchereria bancrofti infections in Haitian schoolchildren. AB - This randomized, placebo-controlled trial investigated the efficacy and nutritional benefit of combining chemotherapeutic treatment for intestinal helminths (albendazole) and lymphatic filariasis (ivermectin). Children were infected with Ascaris (29.2%), Trichuris (42.2%), and hookworm (6.9%), with 54.7% of children having one or more of these parasites. Wuchereria bancrofti microfilaria were found in 13.3% of the children. Children were randomly assigned to treatment with placebo, albendazole, ivermectin, or combined therapy. Combination treatment reduced the prevalence of Trichuris infections significantly more than either drug alone. Combination therapy also significantly reduced the prevalence and density of W. bancrofti microfilaremia compared with placebo or ivermectin alone. Only combination therapy resulted in significantly greater gains in height (hookworm-infected children) or weight (Trichuris infected children) compared with the placebo group. Combined albendazole and ivermectin was a more efficacious treatment for intestinal helminth and W. bancrofti infections in children and resulted in nutritional benefits not found with either drug alone. PMID- 10466982 TI - Blockage of skin invasion by schistosome cercariae by serine protease inhibitors. AB - Invasion of skin by schistosome cercariae is facilitated by a serine protease secreted from the acetabular cells of cercariae in response to skin lipid. Specific inhibitors of the protease, when applied to human skin in formulations designed to retain the inhibitor on and in the upper stratum corneum layers, block cercarial invasion of human skin. Both peptide-based, irreversible inhibitors and non-peptide, reversible inhibitors block cercarial invasion when applied in a propylene glycol:isopropyl alcohol (3:1) formulation in vitro. Arrest of cercarial invasion could be achieved even after immersion of treated skin in water for 2 hr. Peptide-based irreversible inhibitors in the presence of three different Topicare Delivery Compounds optimized arrest of cercarial invasion. The three Topicare Delivery Compounds applied alone prevented 80-100% of cercarial invasion. With inclusion of the inhibitor, there was 97-100% inhibition in vitro. The optimal formulation with inhibitor was then applied to the tails of BALB/c mice, and the mice were exposed to 120 cercariae by tail immersion. With the carrier lotion alone, there was a 50% reduction in worm burden and a 70% reduction in egg burden. When inhibitor was included, an 80% reduction in worm burden and a 92% reduction in egg burden was observed. PMID- 10466983 TI - Immunochemical characterization and diagnostic potential of a 63-kilodalton Schistosoma antigen. AB - Schistosoma circulating antigens were used for the detection of active infection. Anti-S. mansoni IgG2a monoclonal antibody (MAb) designated C5C4 was generated. The target epitope of this MAb was detected in adult worms, eggs, and cercariae antigenic extracts of S. mansoni and S. haematobium, had a molecular size of 63 kD, and was not detected in Fasciola hepatica and Ascaris. In addition, a 50-kD degradation product was identified only in the urine of infected individuals. Analysis by high-performance liquid chromatography of the purified antigen demonstrated only one peak. The 63-kD antigen was characterized as a protein containing 40.4% hydrophobic, 7.5% acidic, and 8.8% basic amino acids. The C5C4 MAb was used in a Fast Dot-ELISA for rapid and simple diagnosis of human schistosomiasis. The 63-kD circulating antigen was detected in 92% of urine samples from 330 S. mansoni-infected individuals, with 16% false-positive results among 130 noninfected individuals. PMID- 10466984 TI - Co-agglutination test for the detection of circulating antigen in amebic liver abscess. AB - We report here a simple and economical slide agglutination test, the co agglutination (Co-A) test, for the detection of circulating amebic antigen in sera for the diagnosis of amebic liver abscess. Fifty serum specimens from cases of amebic liver abscess, 25 from other individuals with parasitic and miscellaneous infections, and 25 from healthy controls were tested for the presence of serum antigen by the Co-A test. Forty-five (90%) amebic liver abscess sera were found to be amebic-antigen positive by the Co-A test. None of 25 sera from healthy controls were positive for the antigen. However, false-positive results were seen with two sera from those with other parasitic and miscellaneous infection controls. These results show that the Co-A test can be used as a sensitive and specific rapid slide agglutination test for the detection of amebic antigen in the sera for diagnosis of cases of amebic liver abscess in a routine parasitology laboratory. PMID- 10466985 TI - Laboratory investigation of human deaths from vampire bat rabies in Peru. AB - In the spring of 1996, multiple cases of an acute febrile illness resulting in several deaths in remote locations in Peru were reported to the Centers for Disease Control and Prevention (CDC). The clinical syndromes for these cases included dysphagia and encephalitis. Because bat bites were a common occurrence in the affected areas, the initial clinical diagnosis was rabies. However, rabies was discounted primarily because of reported patient recovery. Samples of brain tissue from two of the fatal cases were received at CDC for laboratory confirmation of the rabies diagnosis. An extensive array of tests on the formalin fixed tissues confirmed the presence of both rabies viral antigen and nucleic acid. The virus was shown to be most closely related to a vampire bat rabies isolate. These results indicate the importance of maintaining rabies in the differential diagnosis of acute febrile encephalitis, particularly in areas where exposure to vampire bats may occur. PMID- 10466986 TI - Estimates of gene flow among Anopheles maculatus populations in Thailand using microsatellite analysis. AB - We report an analysis of seven microsatellite loci in eight populations of Anopheles maculatus mosquitoes dispersed over a distance of approximately 1,100 km in Thailand. A wide spectrum of genetic variability, with mean heterozygosities ranging from 0.738 to 0.847 were found. Based on microsatellite analysis, geographic populations of An. maculatus can be grouped into two clusters; one includes upper and lower northern populations that extend from approximately 11 degrees to 16 degrees north latitude, and the other (southern populations) extends south from about 7 degrees to 6 degrees north latitude. Wright's F(ST) and Slatkins's R(ST) for all seven microsatellite loci indicated low estimates of differentiation among all populations (mean values of F(ST) and R(ST) = 0.0406 and 0.051, respectively, corresponding to the Nm values of 5.91 and 4.65, respectively), and suggested that gene flow occurs among populations. However, there is some restriction of gene flow between the northern and southern populations. Geographic barriers could be limiting factors for greater gene flow between populations. PMID- 10466987 TI - Progress in the management of type I thoracoabdominal and descending thoracic aortic aneurysms. AB - We reviewed our categorization of patients at high risk for neurologic complications in the repair of descending thoracic and thoracoabdominal aortic aneurysm in which we used cerebrospinal fluid drainage and distal aortic perfusion (adjuncts). A total of 409 patients were operated on by one surgeon for descending thoracic or thoracoabdominal aortic aneurysm between 1992 and 1997. Of these patients, 232 had total descending thoracic or type I thoracoabdominal aortic aneurysm, 131 (56%) of whom were operated on with adjuncts. These patients were compared to 101 nonadjunct patients for demographic variables, intraoperative variables, blood product consumption, and neurologic status. In 131 consecutive patients with adjuncts, all but one awoke from anesthesia without neurologic deficit. In nonadjunct patients, however, neurologic deficit occurred in 6 of 101 (6%) (p < 0.003). The adjunct group had more preoperative renal insufficiency (p < 0.05), an established risk factor for neurologic deficit (odds ratio = 2.2 in published studies). All other risk factors for neurologic deficit occurred with comparable frequency in both groups. We conclude that the introduction of adjuncts has dramatically reduced the neurologic risk associated with type I thoracoabdominal or total descending thoracic aortic repair. Previously considered high risk for neurologic complications, these aneurysms can now be reclassified as low risk in surgery accompanied by adjuncts. Future investigations will focus on type II thoracoabdominal aortic aneurysm as the major source of neurologic morbidity. PMID- 10466988 TI - Early carotid endarterectomy in selected stroke patients. AB - Although there are several reports suggesting the safety of performing carotid endarterectomy (CE) within 4 weeks (early) of a nondisabling stroke, at many institutions it is not standard practice. Benefits of early surgery may include reduction in the number of strokes or carotid occlusions during the time between stroke and surgery, as well as a reduction in the cost of medical care due to the elimination of interval anticoagulation and close follow-up. This review examines the outcomes of early CE in selected patients after a nondisabling stroke. A total of 1065 CEs were performed between November 1991 and April 1998. Seventy five patients were identified by computerized hospital record and office chart review as having CE after a nondisabling stroke. Criteria for early surgery included 1) nondisabling stroke ipsilateral to a carotid stenosis >50%, 2) neurological stability, and 3) no evidence of hemorrhagic stroke or significant cerebral edema by CT/MRI evaluation. This review suggests that early CE can be performed in selected patients with an acceptable perioperative morbidity and mortality. PMID- 10466989 TI - Cerebral ischemic disease and morphometric analyses of carotid plaques. AB - Atherosclerotic carotid plaque morphology and especially, intraplaque hemorrhage are assumed to be related to neurological symptoms. Most researchers have only investigated the incidence of intraplaque hemorrhage in symptomatic and asymptomatic patients. In the present study, the amount of intraplaque hemorrhage is determined in carotid endarterectomy specimens from 33 symptomatic and 14 asymptomatic patients that caused >70% luminal stenosis. The plaque components (fibrosis, lipids, intraplaque hemorrhage, calcification, and intraluminal thrombosis) were quantified as a percentage of the total plaque volume. A high incidence of intraplaque hemorrhage was found in both the symptomatic (94%, 31/33) and asymptomatic (71%, 10/14) patients. The amount of intraplaque hemorrhage was very small within the plaques of the symptomatic (0.39% +/- 0.70%) and asymptomatic (0.37% +/- 1.12%) patients. The plaques of the symptomatic patients contained more fibrosis than lipids (45.62% +/- 14.99% and 20.45% +/- 21.45%, respectively), as did the plaques of the asymptomatic patients (42. 51% +/- 15.28% and 15.46% +/- 15.22%, respectively). Finally, intraluminal thrombosis and calcification were rare. We conclude that the amount of intraplaque hemorrhage was very small and therefore question its direct role in the development of neurological symptoms. In general, the "unstable" plaque contained more fibrosis than lipids. PMID- 10466990 TI - Balloon angioplasty induces heat shock protein 70 in human blood vessels. AB - Balloon angioplasty produces a mechanically induced injury to the blood vessel wall. Heat shock protein 70 (HSP70) is a molecular chaperone whose expression can be induced by chemical or thermal stress. Thus, we hypothesized that the mechanical injury associated with balloon angioplasty would lead to increases in the expression of HSP70 in vascular smooth muscle. Segments of popliteal and trifurcation vessels from above-the-knee amputations were subject to transluminal balloon angioplasty, excised, and placed in organ cultures. Neighboring vessel that was not subjected to balloon angioplasty served as controls. Some vessels were treated with sodium arsenite (positive control, known to induce HSP70 expression). The vessels were homogenized and the proteins were separated by gel electrophoresis and transferred to Immobilon. Western blots with an antibody specific for the inducible form of HSP70 were analyzed by densitometry. Our results showed that HSP70 expression can be induced by the mechanical injury associated with balloon angioplasty in human atherosclerotic vessels. PMID- 10466991 TI - Cholesterol, but not cigarette smoke, decreases rabbit carotid artery relaxation. AB - The purpose of this study was to determine the physiologic effects of cigarette smoke exposure and dietary cholesterol on the availability of nitric oxide in carotid vascular rings. New Zealand white rabbits were placed in an airflow chamber for 3 hr/day over an 8-week period and were exposed to smoke from 600 cigarettes/per day added to the chamber inflow by a robotic smoke generator. New Zealand white rabbits, made hypercholesterolemic, and one group fed a normal diet, were similarly placed in the chamber without exposure to cigarette smoke. In those exposed groups, serum cotinine and cholesterol levels were consistently elevated. After the 8-week period, the carotid arteries were harvested. The vessels were cut into 3-mm rings which were suspended from pressure transducers. The rings were contracted with potassium chloride (KCl) to determine vessel integrity. One ring from each carotid was maximally contracted with 1 x 10(-3) molar norepinephrine (NE) while the experimental ring was contracted to 50% of maximum. Relaxation of the rings was achieved by adding incremental doses of acetylcholine. Our results showed that endothelial dysfunction, as measured by acetylcholine-mediated vasorelaxation, occurs in the rabbit carotid artery when exposed to high dietary cholesterol. Cigarette exposure alone in this particular vessel did not result in significant alteration in acetylcholine-mediated vasorelaxation. PMID- 10466992 TI - Combination therapy of cholesterol reduction and L-arginine supplementation controls accelerated vein graft atheroma. AB - Hyperlipidemia contributes to the development of intimal hyperplasia and accelerated atheroma in vein bypass grafts. Dietary cholesterol reduction and oral supplementation with L-arginine have been shown to reduce accelerated atheroma in experimental vein grafts. This study extends these observations by examining the effect of the combination therapy of cholesterol reduction and L arginine supplementation on the development of intimal hyperplasia in vein grafts in hypercholesterolemic animals. Thirty New Zealand White rabbits had a carotid vein bypass graft performed and were sacrificed at 28 days postoperatively either for morphology (light and electron microscopy) and videomorphometry, or for in vitro contractile studies. Twenty animals received a 1% cholesterol diet for 4 weeks prior to surgery. This diet was continued until harvest in ten animals. Ten cholesterol-fed animals received L-arginine supplementation (2 g/kg/day, p.o.) for 7 days preoperatively and thereafter until harvest and in addition were returned to a normal diet on the day of surgery. The last ten animals were controls (normal diet). Combined cholesterol reduction and L-arginine supplementation prevented accelerated atheroma in vein grafts, halted the change in enhanced smooth muscle cell contractility, and improved endothelial cell function. Early postoperative therapy targeting atheroma development in the high risk patient could offer significant morphological and functional benefits. PMID- 10466993 TI - The role of duplex scanning in the diagnosis of lower limb arterial disease. AB - Color flow duplex imaging of the iliac and femoropopliteal arteries was performed in patients undergoing angiography. The aim of the study was to determine: (1) in what percentage of patients could the iliac arteries be adequately visualized to enable a diagnosis, (2) the overall accuracy of duplex scanning in the diagnosis of arterial disease, and (3) whether there is a useful duplex criterion for the selection of patients for angioplasty. One hundred and twenty patients (79 males, 41 females; mean age 64.4 years) had duplex scans prior to angiography (2-7 days) and the results were compared. The duplex criteria of an increase in the peak systolic velocity ratio (PSVR) >2 and lesions <5 cm were used to signify hemodynamically significant stenosis (>50% narrowing), the presence of plaque and calcification in the arterial wall with alteration of PSVR and lesions >5 cm, diffuse disease, and the absence of flow on color/Doppler interrogation, occlusion. The results show that duplex scanning is a useful screening tool and may be effectively used to diagnose iliac and femoropopliteal disease in nearly 80% of patients. Angiography will be needed in those in whom duplex scanning is inconclusive, or, prior to intervention in those with disease suitable for surgical reconstruction or angioplasty, diagnosed on the basis of duplex scans. PMID- 10466994 TI - Assessment of cardiac risk before aortic reconstruction: noninvasive work-up using clinical examination, exercise testing, and dobutamine stress echocardiography versus routine coronary arteriography. AB - In this prospective study we evaluated the efficacy of a battery of noninvasive tests including clinical evaluation (CE), exercise testing (ET), and dobutamine stress echocardiography (DSE) for assessment of cardiac risk in 90 patients indicated for aortic reconstruction. As the gold-standard reference technique, coronary arteriography was performed in each patient after noninvasive evaluation. The sensitivity of CE was low (61%). ET proved to be more sensitive (71.4%) and highly specific (95.8%) but feasibility (77%) and diagnostic accuracy (42%) were low. DSE demonstrated acceptable sensitivity (78%) and specificity (75.5%) with high feasibility (94.5%) and diagnostic accuracy (100%). None of the four patients with false negative ET results and only one of seven with false negative DSE required coronary bypass. On the basis of these findings we conclude that a combination of CE and ET with DES, if necessary, can reliably assess cardiac risk before aortic reconstruction. Noninvasive assessment is a reliable alternative to routine coronary arteriography. PMID- 10466995 TI - Impregnated polyester arterial prostheses: performance and prospects. AB - Impregnated polyester arterial prostheses have gained wide acceptance by most vascular surgery teams, probably because these prostheses are easy to use, without any preclotting. We offer here a synthesis of the main studies that have appraised the experimental and clinical performance of these prostheses, and we delineate their major prospects. PMID- 10466996 TI - Results of valvuloplasty in patients presenting deep venous insufficiency and recurring ulceration. AB - The purpose of this retrospective study was to assess mid-term results of valvuloplasty in patients presenting chronic recurring venous stasis ulceration. From 1988 to 1993, valvuloplasty was performed in the superficial femoral vein of 33 lower extremities in 28 patients presenting recurring ulceration. In 23 cases, previous surgery in the superficial venous system or perforating vein had failed. Preoperative work-up demonstrated primary deep venous insufficiency (PDVI) in 22 extremities (group I), proximal PDVI in association with distal postthrombotic syndrome (PTS) in 10 (group II), and Klippel-Trenaunay syndrome in 1. Hemodynamic assessment with tourniquet placement demonstrated a mean venous return time of 9 sec (+10, -8). Descending femoral phlebography showed Kistner grade 4 in 30 cases. Outcome was evaluated by clinical examination and Dupplex scan with photophlethysmography at follow-up times ranging from 2 to 7.6 years (mean: 51 months). Correlation between outcome of valvuloplasty and clinical findings was excellent. The incidence of poor clinical and hemodynamic results was higher for patients with PTS. Valve repair in association with surgery for superficial vein insufficiency and ligation of perforators gives good results in patients with isolated PDVI. PMID- 10466997 TI - Is routine postaneurysmectomy hemodynamic assessment of the inferior mesenteric artery circulation helpful? AB - All patients with an abdominal aortic aneurysm treated during a 27-year period by one surgical group at the MidAmerica Heart Institute were included in this study. A prospective routine postaneurysmectomy hemodynamic assessment of the inferior mesenteric artery (IMA) circulation was performed in a test group of consecutive patients operated on by one surgeon. When a mean IMA stump pressure 48 months. This case is the first example of arterial placement of a vena cava filter. This report describes the probable mechanisms for this aberrant placement, methods for prevention of this complication, and options for management of this problem. PMID- 10467000 TI - Isolated hypogastric artery aneurysms. AB - Iliac artery aneurysms are rare in the absence of concomitant abdominal aortic aneurysm (AAA), and isolated internal iliac (hypogastric) aneurysms in particular are extremely rare. From 1986 to 1997 we repaired 572 aortic and/or iliac artery aneurysms in 440 patients. Among these there were only seven hypogastric aneurysms and three of these occurred in the absence of, or remote to, AAA. Hypogastric aneurysms are difficult to diagnose, and large aneurysms are associated with significant morbidity and mortality due to compression of adjacent structures and a high rate of rupture. They pose technical challenges in repair because of their location deep in the pelvis and because it is difficult to gain distal control of the hypogastric artery and its branches. However, the technique of obliterative endoaneurysmorrhaphy has made repair of these aneurysms safe and straightforward. Moreover, this method, unlike percutaneous endovascular techniques, eliminates the compressive mass that is often associated with significant symptomatology. We report three isolated hypogastric aneurysms repaired over an 11-year period, illustrating the technique of proximal ligation and obliterative endoaneurysmorrhaphy, and review the literature on the topic. PMID- 10467001 TI - Controlling septation in fission yeast: finding the middle, and timing it right. AB - The fission yeast Schizosaccharomyces pombe provides a simple eukaryotic model for the study of cytokinesis. S. pombe cells are rod-shaped, grow mainly by elongation at their tips, and divide by binary fission after forming a centrally placed division septum. Analysis of mutants has begun to shed light upon how septum formation and cytokinesis are regulated both spatially and temporally. Some of the proteins involved in these events have been functionally conserved throughout eukaryotic evolution, suggesting that aspects of this control will be common to all eukaryotic cells. PMID- 10467002 TI - Gene ste20 controls amiloride sensitivity and fertility in Schizosaccharomyces pombe. AB - It has been shown previously that amiloride, a widely used diuretic drug, inhibits growth in Schizosaccharomyces pombe. Here we show that the drug also alleviates repression by various nutrients of mating and sporulation in fission yeast. We selected spontaneous mutants that are amiloride-resistant and unable to mate and sporulate. One of them defines the gene ste20. This gene has been cloned and sequenced. It codes for a putative protein of 1309 amino acids. Its sequence does not provide any clues to its function. In contrast to the wild-type, mutants defective in this gene can grow in a medium containing 40 microm amiloride, do not arrest in G(1), and do not induce ste11 expression upon nitrogen starvation and thus are sterile. In addition the ste20 mutants are methylamine-sensitive, exhibit enhanced medium acidification and are defective in the utilization of gycerol as a carbon source. PMID- 10467003 TI - The pub1 E3 ubiquitin ligase negatively regulates leucine uptake in response to NH(4)(+) in fission yeast. AB - Fission yeast strains auxotrophic for leucine are unable to proliferate in normally supplemented minimal media adjusted to pH 6. 4 or above. High-pH sensitivity can be suppressed by the loss of Pub1, an E3 ubiquitin ligase, or by the replacement of NH(4)(+) with a non-repressing source of nitrogen such as L proline. In this report we show pub1 to be required for the rapid down-regulation of leucine uptake observed in response to the addition of NH(4)(+) to the growth media. Furthermore, we corroborate earlier results demonstrating the transport of leucine to be negatively influenced by high extracellular pH. pub1 is homologous to the budding yeast nitrogen permease inactivator, NPI1/RSP5, which mediates the ubiquitination and subsequent destruction of NH(4)(+)-sensitive permeases. The high-pH sensitivity of cells auxotrophic for leucine thus seems to reflect an inability of NH(4)(+)-insensitive permeases to transport sufficient leucine under conditions where the proton gradient driving nutrient transport is low, and NH(4)(+)-sensitive permeases have been destroyed. Intriguingly, the partial suppression of both high pH sensitivity, and the inactivating effect of NH(4)(+) on leucine transport, seen in pub1-1 point mutants, becomes as complete as seen in pub1Delta backgrounds when cells have concomitantly lost the function of the spc1 stress-activated MAPK. PMID- 10467004 TI - A novel group-II intron in the cox1 gene of the fission yeast Schizosaccharomyces pombe is inserted in the same codon as the mobile group-II intron aI2 in the Saccharomyces cerevisiae cox1 homologue. AB - We describe herein a large group-II intron which is inserted in the mitochondrial cox1 gene of the Schizosaccharomyces pombe strain EF2. The intron RNA consists of 2492 nucleotides which can be folded into a secondary structure with all the expected sequence motifs of subgroup-IIA1 introns (Michel et al. 1989). Determination of the exact splice point revealed that the intron is inserted in the same codon, but 1 bp downstream, as the mobile intron aI2 in the Saccharomyces cerevisiae cox1 homologue. A total of nine nucleotide changes was observed around the insertion site of the intron in the cox1 gene of strain EF2 compared with the reference strain ade7-50h(-). Seven of these changes are clustered within the 51 bp upstream of the splice point. Only one sequence deviation was found in the downstream exon. The intron is capable of splicing despite the fact that both the EBS1/IBS1 and the EBS2/IBS2 sequence motifs, thought to be necessary for correct splicing, extend over 5 instead of 6 bp. The maturase, endonuclease and reverse transcriptase domains of the putative protein encoded by the newly described S. pombe group-II intron were not closer to those encoded by the other two, cobI and cox2I, S. pombe group-II introns than to the group-II intron-encoded proteins in Allomyces, Marchantia, Podospora and Saccharomyces. PMID- 10467005 TI - Cysteine uptake by Saccharomyces cerevisiae is accomplished by multiple permeases. AB - Uptake by Saccharomyces cerevisiae of the sulphur-containing amino acid L cysteine was found to be non-saturable under various conditions, and uptake kinetics suggested the existence of two or more transport systems in addition to the general amino-acid permease, Gap1p. Overexpression studies identified BAP2, BAP3, AGP1 and GNP1 as genes encoding transporters of cysteine. Uptake studies with disruption mutants confirmed this, and identified two additional genes for transporters of cysteine, TAT1 and TAT2, both very homologous to BAP2, BAP3, AGP1 and GNP1. While Gap1p and Agp1p appear to be the main cysteine transporters on the non-repressing nitrogen source proline, Bap2p, Bap3p, Tat1p, Tat2p, Agp1p and Gnp1p are all important for cysteine uptake on ammonium-based medium. Furthermore, whereas Bap2p, Bap3p, Tat1p and Tat2p seem most important under amino acid-rich conditions, Agp1p contributes significantly when only ammonium is present, and Gnp1p only contributes under the latter condition. PMID- 10467006 TI - Compact organization of rRNA genes in the filamentous fungus Ashbya gossypii. AB - The rDNA cluster in the phytopathogenic fungus Ashbya gossypii consists of approximately 50 tandem repeat units of 8197 bp. Each unit carries a gene for the 35S pre-rRNA, processed into 18S, 5.8S and 25S rRNA, and a divergently transcribed gene for 5S rRNA. The well-characterized rDNA of the yeast Saccharomyces cerevisiae is the only other example of a completely sequenced rDNA unit (9137 bp) carrying both a 35S pre-rRNA and a 5S rRNA gene. The coding regions for the 5S, 5.8S, 18S and 25S rRNAs are 95-100% identical whereas transcribed and non-transcribed spacers show 43-66% sequence identity. Functionally characterized rDNA and rRNA elements of S. cerevisiae can be unambiguously recognized in the A. gossypii sequence, including the RNA polymerase-I transcription start site, two Reb1p enhancer binding sites and numerous recognition sequences for rRNA modification and processing. In addition to these functionally characterized sequences eight highly conserved elements from 10 to 71 bp were detected in the over 600-bp transcribed region upstream of the 18S rRNA gene which most likely play as yet uncharacterized functions at the DNA or RNA level. In addition to this work we started to identify A. gossypii homologs of S. cerevisiae nucleolar proteins involved in rDNA maturation. PMID- 10467007 TI - Isolation of a new set of Aspergillus nidulans mutants defective in nuclear migration. AB - In the filamentous fungus Aspergillus nidulans, nuclear migration in the germ tube is mediated by cytoplasmic dynein. We have previously reported the characterization of four nud (nuclear distribution) genes, nudA, nudC, nudF and nudG, involved in this process. The nudA and nudG genes respectively encode for the heavy chain and the 8-kDa light chain of cytoplasmic dynein. In this work, we describe an improved method for the isolation of nud mutants that has led to the identification of at least ten additional nud loci. We have cloned one of the genes, nudK, and determined that it encodes the actin-related protein Arp1, which is a component of the dynactin complex. This provides the first evidence that dynactin is involved in nuclear migration in A. nidulans. PMID- 10467008 TI - Polymorphism around cog extends into adjacent structural genes. AB - The recombination hotspot cog overlaps a highly polymorphic 950-bp region of linkage group I in Neurospora crassa. The sequence of this region in the four strains, Lindegren 25a, Lindegren A, Emerson A and St. Lawrence 74A, each differs from the others by 1.4% or more. Comparison of the sequence of St. Lawrence 74A and Lindegren 25a each side of cog shows a high level of sequence heterology extending in both directions, including the coding sequences for his-3 and a putative gene lpl with homology to yeast lysophospholipase. The St. Lawrence 74A and Lindegren 25a sequences of his-3, centromere-proximal to cog, differ at 14 nucleotides, resulting in six amino-acid variations between the predicted protein sequences. In lpl, distal from cog, the sequences differ at 19 nucleotides leading to five amino-acid differences between the predicted proteins. Sequence heterology between St. Lawrence 74A and Lindegren 25a peaks either side of cog and then declines with distance. At the am locus on linkage group V, heterology is much less but peaks close to a weak recombination hotspot 5' of the coding sequence. Uneven distribution of polymorphism along chromosomes has been explained by a hitch-hiking hypothesis in which selection for advantageous mutations causes local fixation of unselected variation. We suggest that new mutations arising from errors in recombination also contribute to the uneven distribution of polymorphism. PMID- 10467009 TI - The metG gene of Aspergillus nidulans encoding cystathionine beta-lyase: cloning and analysis. AB - The metG gene of Aspergillus nidulans encoding cystathionine beta-lyase, an enzyme of the main pathway of methionine synthesis, was cloned by complementation of a metG mutation. A comparison of metG genomic DNA and a cDNA copy derived from different A. nidulans strains revealed a marked DNA sequence polymorphism manifested mostly by silent point mutations. cDNA of the A. nidulans metG gene complemented the Escherichia coli metC69 mutation impairing cystathionine beta lyase. This gene contains two introns and codes for a protein of 439 amino acids. The protein shows homology with bacterial, yeast and plant cystathionine beta lyases, as well as with other enzymes belonging to a large family of pyridoxal 5' phosphate binding proteins. Transcription of the metG gene is not appreciably regulated by the concentration of sulphur source in the growth medium. PMID- 10467010 TI - Molecular karyotypes for Alternaria plant pathogens known to produce host specific toxins. AB - There are at least ten plant diseases caused by Alternaria species in which host specific toxins (HSTs) are responsible for fungal pathogenicity. Of these HST producers, seven are considered distinct pathotypes of the species Alternaria alternata, and the remaining three are among other species of pathogenic Alternaria. Inter- and intra-specific variation among Alternaria taxa, including HST-producers, was determined by electrophoretic karyotyping using pulsed-field gel electrophoresis. A. alternata including seven pathotypes of A. alternata and eight non-pathogenic strains had 9-11 chromosomal bands with estimated sizes ranging from 0.4 to 5.7 Mb. In contrast, Alternaria species that are morphologically distinct from A. alternata had 8-10 bands with sizes between 0.9 and 5.7 Mb. Estimated genome sizes of A. alternata and other Alternaria species ranged from 28.8 to 33.6 Mb and 25.1 to 30.7 Mb, respectively. Other species of pathogenic Alternaria were difficult to differentiate from A. alternata on the basis of chromosome-size polymorphisms alone, but Southern analysis using rDNA as a probe could, in some cases, differentiate between them. These results were cytologically confirmed by 4',6-diamidino-2-phenylindole (DAPI) staining and fluorescence in situ hybridization with a rDNA probe for mitotic metaphase chromosomes prepared by the germ-tube burst method. PMID- 10467012 TI - Changes of Respiration Rate, Ethylene Evolution, and Abscisic Acid Content in Developing Inflorescence and Young Fruit of Olive (Olea europaea L. cv. Konservolia). AB - Simultaneous measurements of respiration, ethylene production, and abscisic acid (ABA) concentrations, as well as the growth parameters length, fresh weight (FW), and dry weight (DW) of olive (Olea europaea L. cv. Konservolia) inflorescence were carried out at short intervals (3-7 days) during the period from bud burst until the 3rd week after full bloom (AFB), when young fruit reached 8 mm in length. The axis of inflorescence elongated remarkably during the 3rd week after bud burst (ABB), massive bract shedding occurred during the 4th week ABB, full bloom (FB) was observed 7 weeks ABB, and massive floral organ abscission 1 week AFB. The results showed a continuous increase in inflorescence FW and DW from bud burst until 4 days before FB. Respiration rate, ethylene production, and levels of ABA were relatively high during the first 3 weeks ABB. After this period, respiration and ethylene followed a similar pattern of changes, inversely to that of ABA concentration. An accumulation of inflorescence ABA 6 and 4 days before FB was associated with the minimum values of respiration and ethylene production on the same dates. The sharp decrease in the ABA concentration during FB and 3 days later was followed by a high rise in ethylene and an increase in respiration rate, which both rose further 1 week AFB. The results suggest a possible correlation of ABA with the early stage of floral abscission, whereas ethylene production seems to be correlated with the terminal separatory activity in olive inflorescence abscission processes.Key Words. Abscisic acid-Abscission-Ethylene Inflorescence-Olive-Respirationhttp://link.springer ny.com/link/service/journals/00344/bibs/18n1p1.html PMID- 10467011 TI - Expression of a higher-plant chloroplast psbD promoter in a cyanobacterium (Synechococcus sp. strain PCC7942) reveals a conserved cis-element, designated PGT, that differentially interacts with sequence-specific binding factors during leaf development. AB - The chloroplast psbD gene, which encodes the D2 subunit of photosystem II, is regulated by a blue light-responsive promoter (BLRP). We tested the ability of different regions of the barley (Hordeum vulagare) BLRP to drive transcription of the lacZ reporter gene in genomic transformants of Synechococcus sp. strain PCC7942. The barley BLRP was transcribed in Synechococcus from the same initiation sites that are used in plant chloroplasts in vivo. A region of the BLRP, residing between -83 and -112 bp upstream from the transcription initiation sites, functioned as a negative element in Synechococcus. Nucleotide sequences within this region are conserved among the psbD genes of several monocots and dicots, and with the nuclear negative regulatory element GT. Thus this new cis element was designated Plastid GT, PGT. Proteins from chloroplasts of barley and Arabidopsis thaliana interacted with PGT in a sequence-specific and developmental dependent manner. The DNA-protein complexes from Arabidopsis chloroplasts are composed of 60- and 38-kDa polypeptides. We postulate that GT and PGT have evolved in the nucleus and chloroplast, respectively, from a common ancestral regulatory element. PMID- 10467014 TI - Alleviation of Waterlogging Damage in Winter Rape by Uniconazole Application: Effects on Enzyme Activity, Lipid Peroxidation, and Membrane Integrity. AB - Oilseed rape (Brassica napus L.) seedlings treated with uniconazole [(E)-1-(4 chlorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-l-yl)-l-penten-3-ol] were transplanted at the five-leaf stage into specially designed experimental containers and then exposed to waterlogging for 3 weeks. After waterlogging stress, uniconazole-treated seedlings had significantly higher activities of superoxide dismutase, catalase, and peroxidase enzymes and endogenous free proline content at both the seedling and flowering stages. Uniconazole plus waterlogging-treated plants had a significantly higher content of unsaturated fatty acids than the waterlogged plants. There was a parallel increase in the lipid peroxidation level and electrolyte leakage rate from the leaves of waterlogged plants. Leaves from uniconazole plus waterlogging-treated plants had a significantly lower lipid peroxidation level and electrolyte leakage rate compared with waterlogged plants at both the seedling and flowering stages. Pretreatment of seedlings with uniconazole could effectively delay stress-induced degradation of chlorophyll and reduction of root oxidizability. Uniconazole did not alter the soluble sugar content of leaves and stems, after waterlogging of seedlings. Uniconazole improved waterlogged plant performance and increased seed yield, possibly because of improved antioxidation defense mechanisms, and it retarded lipid peroxidation and membrane deterioration of plants.Key Words. Waterlogging-Uniconazole-Brassica napus L.-Enzymes-Lipid peroxidation-Membrane integrityhttp://link.springer-ny.com/link/service/journals/00344/bibs/18n1p9.html PMID- 10467015 TI - Early Application of Ethrel Extends Tomato Fruit Cell Division and Increases Fruit Size and Yield with Ripening Delay. AB - Flowers of tomato (Lycopersicon esculentum Mill.) plants cv. Castle Rock were sprayed with 100 ppm of ethrel, 0.5 mm aminooxyacetic acid (AOA), or water (control) 2 days after anthesis. The fruit period of cell division was extended up to 16-18 days after anthesis with the application of ethrel but reduced from 10-12 days (control) down to only 6-8 days with the application of AOA. In a trend opposite to AOA application, fruits that received ethrel treatment were of higher ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC) levels than control. This was noticed not only during the first 2 weeks after anthesis but also during the fruit climacteric phase. Mesocarp cells of ethrel-treated fruits were greater in number/mm(2) but smaller in size than control; an opposite trend was obtained with the application of AOA. This was observed for a period of 18 days after anthesis, but by that time or at earlier ages, fruits of AOA treatment were larger in size and heavier in weight than control, and both were larger and heavier than ethrel-treated ones. At 5 weeks after anthesis and thereafter, the fruit response to all treatments was totally reversed because early ethrel treated fruits became significantly larger in size and heavier in weight with a ripening delay of about 10 and 15 days compared with those of control and AOA treated ones, respectively. When the same treatments were applied to the whole plant, similar results were obtained because the early application of ethrel increased the fruit yield by about 15% over control with a pronounced ripening delay; an opposite trend was obtained with the application of AOA. No significant differences were found among all treatments in terms of flower or fruit abscission or fruit number/plant. The data suggest that ethylene regulates tomato fruit transmission from cell division to cell enlargement. In addition, fruit cell division is terminated only when endogenous ethylene decreases to its basal level, allowing cell enlargement to dominate and proceed as in the case of the early application of AOA. The ripening delay of ethrel-treated fruits may be caused by the longer time required for the increased cell number to reach maturation. A low level of ethrel application at the tomato early fruiting stage may be used for increasing fruit yield by increasing fruit size and consequently its quality.Key Words. Tomato-Ethrel-AOA-Cell division-Cell enlargement-Ethylene ACC-Ripeninghttp://link.springer ny.com/link/service/journals/00344/bibs/18n1p15.html PMID- 10467016 TI - Indole-3-Acetic Acid Control on Acidic Oat Cell Wall Peroxidases. AB - Incubation of oat coleoptile segments with 40 um indoleacetic acid (IAA) induced a decrease of 35-60% in peroxidase activity at the cell wall compartment. Treatment with IAA also produced a similar decrease in the oxidation of NADH and IAA at the cell wall. Isoelectric focusing of ionic, covalent, and intercellular wall peroxidase fractions showed that acidic isoforms (pI 4.0-5.5) were reduced preferentially by IAA treatment. Marked differences were found between acidic and basic wall isoperoxidases in relation to their efficacy in the oxidation of IAA. A peroxidase fraction containing acidic isoforms oxidized IAA with a V(max)/s(0.5) value of 2.4 x 10(-2) min(-1). g fw(-1), 4.0 times higher than that obtained for basic peroxidase isoforms (0.6 x 10(-2) min(-1). g fw(-1)). In contrast, basic isoforms were more efficient than acidic isoperoxidases in the oxidation of coniferyl alcohol or ferulic acid with H(2)O(2) (5.6 and 2.1 times, respectively). The levels of diferulate and lignin in the walls of oat coleoptile segments were not altered by treatment with IAA. The decrease in cell wall peroxidase activity by IAA was related more to reduced oxidative degradation of the hormone than to covalent cell wall cross-linking.Key Words. IAA-Auxin-Wall peroxidases-Oat-Avena sativa-Diferulic acid-Ligninhttp://link.springer ny.com/link/service/journals/00344/bibs/18n1p25.html PMID- 10467017 TI - Influence of Polyamines on Growth of Hairy Root Cultures of Witloof Chicory (Cichorium intybus L. cv. Lucknow Local) and Formation of Coumarins. AB - The effect of polyamines (putrescine, spermidine, and spermine) was examined for growth and production of two coumarins, esculetin and esculin, in the hairy roots of chicory (Cichorium intybus L. cv. Lucknow local). Of the polyamines administered, 1.5 mm putrescine alone resulted in a 2.3-fold higher increase in the growth of hairy roots as well as in the production of esculetin and esculin, which was 3.37 times more than that of the control on day 21. The endogenous level of conjugated putrescine was more than fivefold that of free putrescine levels in untreated samples. The production of esculetin and esculin in hairy root cultures strictly correlated with growth in all of the treatments. Putrescine at 1.5 mm resulted in a greater length of primary root (18.29 +/- 1.37 cm) compared with the control (10.96 +/- 0.82 cm) and more secondary and tertiary roots. This study also provides insight into the morphogenetic changes that occur in roots in response to the external supply of polyamines.Key Words. Cichorium intybus L. cv. Lucknow local-Hairy root cultures-Polyamines Coumarinshttp://link.springer ny.com/link/service/journals/00344/bibs/18n1p33.html PMID- 10467018 TI - Effect of Bis(guanylhydrazones) on Growth and Polyamine Uptake in Plant Cells. AB - In the present work the effect of several bis(guanylhydrazones) on the growth of Helianthus tuberosus tuber explants was studied. Different aliphatic congeners of glyoxal bis(guanylhydrazone) were tested. Most of the compounds displayed an inhibitory effect on growth, and a correlation between the structure of the molecule and the inhibitory activity was observed. Experiments carried out with glyoxal bis(guanylhydrazone) and its congeners methyl-, ethylmethyl-, and methylpropylglyoxal bis(guanylhydrazones) show that as the total number of side chain carbon atoms in the molecule increases, the inhibitory potency also increases. A depletion of spermidine levels was also found in the explants treated with ethylmethylglyoxal bis(guanylhydrazone), which turned out to be one of the most potent growth inhibitors. The addition of spermidine caused a significant reversion of the antiproliferative action of glyoxal bis(guanylhydrazone). The effect of these compounds on spermidine uptake in protoplasts isolated from carrot phloem parenchyma was also investigated. Only a slight competition was found when antagonists were present at concentrations 20 times higher than the polyamine, thus suggesting that bis(guanylhydrazones) do not share, at least at low concentrations, the polyamine transport system in plant cells.Key Words. Bis(guanylhydrazones)-Carrot protoplasts-Growth-Helianthus tuberosus-Polyamines-Uptakehttp://link.springer ny.com/link/service/journals/00344/bibs/18n1p39.html PMID- 10467019 TI - Conditioning of Fasciation by Gibberellin and Genotype in Cotton (Gossypium hirsutum L.). AB - The round-leafed mutant cotton line L-501 developed fasciation of the upper stem when field grown in Central Asia. Fasciation co-segregated with the mutant gene for round leaves In.(l) Fasciation developed at the flowering stage, but removal of floral buds did not prevent fasciation. Fasciation in L-501 could be inhibited by the gibberellin (GA) biosynthesis inhibitor chlorocholine chloride or by fusicoccin. GA(3) application in the field induced fasciation in the mutant's parental line L-463, which has five-lobed leaves and does not normally develop fasciation. Fasciation did not develop in either line, even after GA(3) treatment, in UK glasshouse conditions.Key Words. Fasciation-Gibberellin Gossypium-Leaf-shape-Mutanthttp://link.springer ny.com/link/service/journals/00344/bibs/18n1p45.html PMID- 10467020 TI - Effect of hyaluronidase on albumin diffusion in lung interstitium. AB - Albumin diffusion measured in an isolated segment of rabbit lung interstitium with a radioactive tracer ((125)I-albumin) technique was independent of albumin concentration and similar to the free diffusion of albumin in water (Qiu et al, 1998. J Appl Physiol 85: 575-583). We studied the effect of hyaluronidase on the diffusion of albumin. Isolated rabbit lungs were inflated with silicon rubber by way of airways and blood vessels, and two chambers were bonded to the sides of a approximately 0.5-cm thick slab enclosing a vessel with an interstitial cuff. One chamber was filled with 2 g/dl albumin solution containing (125)I-albumin and 0.02 g/dl hyaluronidase. Unbound (125)I was removed from the tracer by dialysis before use. The other chamber filled with Ringer's solution was placed within a NaI(Tl) scintillation detector. Diffusion of tracer was measured continuously for 120 h. Albumin diffusion coefficient (D) and interstitial area (A) were obtained by fitting the tracer-time curve with the theoretical solution of the equation describing one-dimension diffusion of a solute across a membrane. D averaged 5.2 x 10(-7) cm(2)/s for albumin diffusion with hyaluronidase, 20% less than that measured previously without hyaluronidase. Hyaluronidase had no effect on A. Results indicated an interaction between albumin and interstitial hyaluronan that was the opposite of the steric effect on albumin excluded volume measured in solution. PMID- 10467021 TI - Regional pleural filtration and absorption measured by fluorescent tracers in rabbits. AB - In prone anesthetized rabbits, we used Evans blue-dyed albumin (EBA) to study regional pleural filtration and FITC dextran to study regional pleural absorption. Evans blue was injected intravenously, and the animals were ventilated for 6 h at either of two levels of ventilation. Postmortem the right rib cage was frozen and thawed before study. EBA fluorescence emitted from the rib cage surface was measured along the cranial-caudal axis near the mid chest with fluorescence videomicroscopy. Fluorescent light intensity increased from the third to the eighth rib in a cyclic fashion, with peaks at the ribs and troughs at the intercostal spaces. This increase was greater at the higher ventilation. Fluorescent images of cross sections of a frozen rib cage verified a cranial caudal gradient in filtration. Fluorescent images of FITC dextran absorbed from the pleural space into the rib cage surface indicated major areas of absorption at the ventral, caudal, and cranial regions adjacent to the lung margins and areas of absorption scattered in the intercostal spaces. Simultaneous measurements of EBA filtration and FITC absorption showed sites of maximal filtration that were different from sites of maximal absorption. Pleural uptake of fluorescent microspheres (2-microm diameter) located lymphatic stomata distributed randomly within clusters in the intercostal spaces and channels of lymphatic lacunae parallel to blood vessels. Diaphragmatic uptake of microspheres was confined mainly to the ventral surface of the central tendon. Visceral pleural absorption was minimal. PMID- 10467023 TI - Atypical adrenoceptor-mediated relaxation of canine pulmonary artery through a cyclic adenosine monophosphate-dependent pathway. AB - To determine whether functional atypical beta-adrenoceptors (beta(3) adrenoceptors) are present in pulmonary vascular smooth muscle, we studied isolated canine pulmonary arterial rings under isometric conditions in vitro. Addition of beta-adrenoceptor agonists produced a concentration-dependent relaxation of noradrenaline-precontracted tissues, a rank order potency being isoproterenol (1) > salbutamol (0.95) > selective beta(3)-adrenoceptor agonists, CL 316243 (0.85), and BRL 37344 (0. 83). A marked desensitization to salbutamol occurred by pretreatment with salbutamol but not with CL 316243. When beta(1) adrenoceptors had been blocked, the relaxant responses to salbutamol were competitively antagonized by the beta(2)-adrenoceptor antagonist ICI 118551 with a pA(2) value of 7.67 +/- 0.21 (mean +/- S.E.), but the response to CL 316243 was weekly antagonized by ICI 118551 only at a high concentration of 10(-5) M, where an apparent pA(2) value was 5. 24. In contrast, cyanopindolol, a nonselective beta-adrenoceptor antagonist, antagonized CL 316243-induced relaxation in a competitive manner with a pA(2) of 6.10 +/- 0.11. This pA(2) value was lower than that when salbutamol was used as an agonist (6.69 +/- 0.14, p < 0.01). Intracellular 3',5'-cyclic adenosine monophosphate (cAMP) levels were increased by CL 316243 in a concentration-dependent fashion, an effect that was not altered by ICI 118551. These results suggest that beta(3)-adrenoceptors may exist in canine pulmonary artery smooth muscle and that stimulation of this atypical receptor causes vasodilation through a cAMP-dependent pathway. PMID- 10467022 TI - Elevation of anti-cytokeratin 19 antibody in sera of the patients with idiopathic pulmonary fibrosis and pulmonary fibrosis associated with collagen vascular disorders. AB - It has been suggested that cytokeratin 19 is expressed in regenerated bronchoepithelial cells in patients with pulmonary fibrosis, and serum cytokeratin 19 fragment is elevated in patients with pulmonary fibrosis. We hypothesized that serum antibodies to cytokeratin 19 may be formed in patients with pulmonary fibrosis. To prove the existence of anti-cytokeratin 19 antibodies in patients' sera, human recombinant cytokeratin 19 was stained with patients' sera by a Western immunoblot. Then, we tried to establish an enzyme-linked immunosorbent assay to quantitate anti-cytokeratin 19 antibody in the sera of patients with idiopathic pulmonary fibrosis (IPF) and pulmonary fibrosis associated with collagen vascular disorders (PF-CVD). We demonstrated the anti cytokeratin 19 antibody in patient' sera by a Western immunoblot. In patients with IPF and PF-CVD, significantly high anti-cytokeratin 19 antibody was demonstrated compared with normal volunteers, patients with chronic bronchitis, and patients with pneumonia. These results suggest that anti-cytokeratin 19 antibody may have played a role in the process of lung injury in pulmonary fibrosis. PMID- 10467024 TI - Cytokeratin fragment 19 (CYFRA 21-1) and carcinoembryonic antigen for early prediction of recurrence of lung adenocarcinoma. AB - Fifty patients with adenocarcinoma of the lung were enrolled in this study, including 20 patients with recurrence and 30 patients without recurrence 1 year after surgery. Serial serum levels of cytokeratin fragment 19 (CYFRA 21-1) and carcinoembryonic antigen (CEA) were measured before the operation and 1 week, 1 month, 3 months, 6 months, 9 months, and 12 months after surgery for the early detection of recurrence. The results revealed that the mean serum values of either CYFRA 21-1 or CEA were significantly higher until 9 and 12 months after surgery in the 20 patients with recurrent adenocarcinoma compared with the 30 patients without recurrent adenocarcinoma. We conclude that CYFRA 21-1 is not a better marker than CEA for early prediction of adenocarcinoma. We conclude that CYFRA 21-1 is not a better marker than CEA for early prediction of adenocarcinoma recurrence in lung. PMID- 10467026 TI - Chemical analysis and immunolocalisation of lignin and suberin in endodermal and hypodermal/rhizodermal cell walls of developing maize (Zea mays L.) primary roots. AB - The composition of suberin and lignin in endodermal cell walls (ECWs) and in rhizodermal/hypodermal cell walls (RHCWs) of developing primary maize (Zea mays L.) roots was analysed after depolymerisation of enzymatically isolated cell wall material. Absolute suberin amounts related to root length significantly increased from primary ECWs (Casparian strips) to secondary ECWs (suberin lamella). During further maturation of the endodermis, reaching the final tertiary developmental state characterised by the deposition of lignified secondary cell walls (u-shaped cell wall deposits), suberin amounts remained constant. Absolute amounts of lignin related to root length constantly increased throughout the change from primary to tertiary ECWs. The suberin of Casparian strips contained high amounts of carboxylic and 2-hydroxy acids, and differed substantially from the suberin of secondary and tertiary ECWs, which was dominated by high contents of omega hydroxycarboxylic and 1,omega-dicarboxylic acids. Furthermore, the chain-length distribution of suberin monomers in primary ECWs ranged from C(16) to C(24), whereas in secondary and tertiary ECWs a shift towards higher chain lengths (C(16) to C(28)) was observed. The lignin composition of Casparian strips (primary ECWs) showed a high syringyl content and was similar to lignin in secondary cell walls of the tertiary ECWs, whereas lignin in secondary ECWs contained higher amounts of p-hydroxyphenyl units. The suberin and lignin compositions of RHCWs rarely changed with increasing root age. However, compared to the suberin in ECWs, where C(16) and C(18) were the most prominent chain lengths, the suberin of RHCWs was dominated by the higher chain lengths (C(24) and C(26)). The composition of RHCW lignin was similar to that of secondary-ECW lignin. Using lignin-specific antibodies, lignin epitopes were indeed found to be located in the Casparian strip. Surprisingly, the mature suberin layers of tertiary ECWs contained comparable amounts of lignin-like epitopes. PMID- 10467027 TI - Isolation and expression analysis of two stress-responsive sucrose-synthase genes from the resurrection plant Craterostigma plantagineum (Hochst.). AB - The enzyme sucrose synthase (UDP-glucose: D-fructose 2alpha-glucosyltransferase, EC 2.4.1.13) is a key enzyme in carbohydrate metabolism, catalyzing the reversible conversion of sucrose uridine-diphosphate into fructose and UDP glucose. We report the molecular characterization of two classes of cDNA and genomic clones encoding sucrose synthase from Craterostigma plantagineum Hochst., a resurrection plant in which the turnover of sucrose is considered to have an important role in the unique phenomenon of surviving desiccation. Sucrose synthase transcript and protein levels are modulated by dehydration and rehydration. In-situ hybridization revealed that transcripts preferentially accumulate in phloem tissues. Promoter analysis underlined a role for class-I sucrose-synthase genes in dehydration stress and in response to cis-abscisic acid. A DNA sequence motif common to class-I sucrose-synthase and sucrose phosphate-synthase genes was discovered. PMID- 10467028 TI - Localization of H(+)-ATPases in soybean root nodules. AB - The localization of H(+)-ATPases in soybean (Glycine max L. cv. Stevens) nodules was investigated using antibodies against both P-type and V-type enzymes. Immunoblots of peribacteroid membrane (PBM) proteins using antibodies against tobacco and Arabidopsis H(+)-ATPases detected a single immunoreactive band at approximately 100 kDa. These antibodies recognized a protein of similar relative molecular mass in the crude microsomal fraction from soybean nodules and uninoculated roots. The amount of this protein was greater in PBM from mature nodules than in younger nodules. Immunolocalization of P-type ATPases using silver enhancement of colloidal-gold labelling at the light-microscopy level showed signal distributed around the periphery of non-infected cells in both the nodule cortex and nodule parenchyma. In the central nitrogen-fixing zone of the nodule, staining was present in both the infected and uninfected cells. Examination of nodule sections using confocal microscopy and fluorescence staining showed an immunofluorescent signal clearly visible around the periphery of individual symbiosomes which appeared as vesicles distributed throughout the infected cells of the central zone. Electron-microscopic examination of immunogold-labelled sections shows that P-type ATPase antigens were present on the PBM of both newly formed, single-bacteroid symbiosomes just released from infection threads, and on the PBM of mature symbiosomes containing two to four bacteroids. Immunogold labelling using antibody against the B-subunit of V-type ATPase from oat failed to detect this protein on symbiosome membranes. Only a very faint signal with this antibody was detected on Western blots of purified PBM. During nodule development, fusion of small symbiosomes to form larger ones containing multiple bacteroids was observed. Fusion was preceded by the formation of cone-like extensions of the PBM, allowing the membrane to make contact with the adjoining membrane of another symbiosome. We conclude that the major H(+) ATPase on the PBM of soybean is a P-type enzyme with homology to other such enzymes in plants. In vivo, this enzyme is likely to play a critical role in the regulation of nutrient exchange between legume and bacteroids. PMID- 10467029 TI - Purification, characterization, and immunolocalization of hydroxycinnamoyl-CoA: tyramine N-(hydroxycinnamoyl)transferase from opium poppy. AB - A development-specific and elicitor-inducible acyltransferase [hydroxycinnamoyl CoA: tyramine N-(hydroxycinnamoyl)transferase (THT; EC 2.3.1.110)] that catalyzes the transfer of hydroxycinnamic acids from hydroxycinnamoyl-CoA esters to hydroxyphenethylamines was purified 988-fold to apparent homogeneity from opium poppy (Papaver somniferum L.) cell-suspension cultures. The purification procedure, which resulted in a 6.8% yield, involved hydrophobic interaction and anion-exchange chromatography, followed by affinity chromatography on Reactive Yellow-3-Agarose using the acyl donor (feruloyl-CoA) as eluent. Purified THT had an isoelectric point of 5.2, a native molecular mass of approximately 50 kDa, and consisted of two apparently identical 25-kDa subunits as determined by two dimensional polyacrylamide gel electrophoresis. The purified enzyme was able to synthesize a variety of amides due to a relatively low specificity for cinnamoyl CoA derivatives and hydroxyphenethylamines. The best substrates were feruloyl-CoA (VK(m)(-1)13.4 mkat g(-1) M(-1)) and tyramine (VK(m)(-1)6.57 mkat g(-1) M(-1)). The THT activity increased during development of opium poppy seedlings, occurred at high levels in roots and stems of mature plants, and was induced in cell suspension cultures after treatment with a pathogen-derived elicitor. Immunoblot analysis using THT mouse polyclonal antibodies did not always show a correlation between THT polypeptide and enzyme activity levels. For example, despite low THT activity in leaves, an abundant 25-kDa immunoreactive polypeptide was detected. Immunohistochemical localization showed that THT polypeptides occur in cortical and xylem parenchyma, immature xylem vessel elements, root periderm, anthers, ovules, and the inner layer of the seed coat, but are most abundant in phloem sieve-tube members in roots, stems, leaves, and anther filaments. PMID- 10467030 TI - Chalcone reductase-homologous transcripts accumulate during development of stem borne nodules on the tropical legume Sesbania rostrata. AB - During a search for genes with induced or enhanced expression in the early stages of development of stem-borne nodules on Sesbania rostrata, a cDNA with homology to chalcone reductase (CHR) genes was isolated. Here, we describe the characterization of a full-length CHR cDNA (Srchr1) and the pattern of CHR transcript accumulation in stem-borne nodules. Expression was correlated with both nodule development and bacterial invasion. In young nodules, CHR transcripts were observed in cells of the parenchyma, in cells around the nodule vascular bundles, and in the uninfected cells of the central tissue. PMID- 10467031 TI - The selective increase or decrease of organellar DNA in generative cells just after pollen mitosis one controls cytoplasmic inheritance. AB - Organellar DNA in mature pollen grains of eight angiosperm species (Actinidia deliciosa Lindl., Antirrhinum majus L., Arabidopsis thaliana (L.) Heynh., Medicago sativa L., Musa acuminata Colla, Pelargonium zonale (L.) L'Her, Petunia hybrida Vilm. and Rhododendron mucronatum (Blume) G. Don, in which the modes of organellar inheritance have been determined genetically, was observed by fluorescence microscopy using Technovit 7100 resin sections double-stained with 4',6-diamidino-2-phenylindole (DAPI) and 3,3'-dihexyloxacarbocyanine iodide (DiOC(6)). The eight species were classified into four types, based on the presence or absence of organellar DNA in mature generative cells: namely (1) type "m+p+", which has both mitochondrial and plastid DNA (P. zonale), (2) type "m+p ", which only has mitochondrial DNA (M. acuminata), (3) type "m-p+", which only has plastid DNA (A. deliciosa, M. sativa, R. mucronatum), and (4) type "m-p-", which has neither mitochondrial nor plastid DNA (A. majus, A. thaliana, P. hybrida). This classification corresponded to the mode of organellar inheritance determined by genetic analysis. The presence or absence of mitochondrial and plastid DNA corresponded to paternal/biparental inheritance or maternal inheritance of the respective organelle, respectively. When organellar DNA was present in mature generative cells (m+ or p+), the DNA content of the organelles in the generative cells started to increase immediately after pollen mitosis one (PMI). In contrast, the DNA content of organelles in generative cells decreased rapidly after PMI when organellar DNA was absent from mature generative cells (m- or p-). These results indicate that the modes of inheritance (paternal/biparental inheritance or maternal inheritance) of mitochondria and plastids are determined independently of each other in young generative cells just after PMI. PMID- 10467032 TI - Molecular characterization of ribulose-1,5-bisphosphate carboxylase/oxygenase activase in rice leaves. AB - A full-length cDNA (designated rcaII) encoding the Rubisco activase (RCA) of rice (Oryza sativa L.) has been cloned from a cDNA library constructed with mRNA from green leaves. Sequence analysis resulted in a reading frame of 432 amino acids with a calculated molecular mass of 47.9 kDa and an estimated isoelectric point of 5.97. The deduced amino acid sequence showed 74-89% identity with other Rubisco activases from higher plants. Two highly conserved motifs were identified. Southern blot analysis suggested the presence of a single rca gene in the rice genome. The accumulation of leaf rca mRNA was found to be regulated by an oscillating circadian rhythm, in rice plants grown in a light-dark photoperiod. To purify the rice RCA protein, total soluble protein from rice green leaves was fractionated by ammonium sulfate precipitation, followed by preparative gel electrophoresis. Two polypeptides, designated RCAI and RCAII, were isolated by two-dimensional gel electrophoresis and further confirmed by N terminal sequencing. The polyclonal antibodies prepared against rice RCAI and RCAII were found to cross-react with two RCA polypeptides present in leaf extracts of spinach and tobacco. Furthermore, two different 3' ends of rca mRNA were detected by reverse transcription-polymerase chain reaction analysis. These cDNA fragments and the related genomic DNA fragment were cloned and sequenced. The sequence of rcaI is almost identical to the corresponding sequence of rcaII, except for its having 33 additional amino acids at the C-terminal portion. It can be concluded that a novel alternative splicing mechanism for a common rca mRNA precursor near the 3' end exists in rice plants. PMID- 10467033 TI - Desiccation and osmotic stress increase the abundance of mRNA of the tonoplast aquaporin BobTIP26-1 in cauliflower cells. AB - Changes in vacuolar structure and the expression at the RNA level of a tonoplast aquaporin (BobTIP26-1) were examined in cauliflower (Brassicaoleracea L. var. botrytis) under water-stress conditions. Gradual drying out of slices of cauliflower floret tissue caused its collapse, with a shrinkage in tissue and cell volumes and an apparent vesiculation of the central vacuole, whereas osmotic stress resulted in plasmolysis with a collapse of the cytoplasm and the central vacuole within. Osmotic stress caused a rapid and substantial increase in BobTIP26 mRNA in slices of floret tissue. Exposure of tissue slices to a regime of desiccation showed a slower but equally large rise in BobTIP26 mRNA followed by a rapid decline upon rehydration. In situ hybridization showed that BobTIP26-2 mRNA is expressed most highly in meristematic and expanding cells of the cauliflower florets and that desiccation strongly increased the expression in those cells and in differentiated cells near the xylem vessels. These data indicate that under water-deficit conditions, expression of the tonoplast aquaporin gene in cauliflower is subject to a precise regulation that can be correlated with important cytological changes in the cells. PMID- 10467034 TI - Tobacco mosaic virus inoculation inhibits wound-induced jasmonic acid-mediated responses within but not between plants. AB - Jasmonic acid (JA) and salicylic acid (SA) have both been implicated as important signal molecules mediating induced defenses of Nicotiana tabacum L. against herbivores and pathogens. Since the application of SA to a wound site can inhibit both wound-induced JA and a defense response that it elicits, namely nicotine production, we determined if tobacco mosaic virus (TMV) inoculation, with its associated endogenous systemic increase in SA, reduces a plant's ability to increase JA and nicotine levels in response to mechanical damage, and evaluated the consequences of these interactions for the amount of tissue removed by a nicotine-tolerant herbivore, Manduca sexta. Additionally, we determined whether the release of volatile methyl salicylic acid (MeSA) from inoculated plants can reduce wound-induced JA and nicotine responses in uninoculated plants sharing the same chamber. The TMV-inoculated plants, though capable of inducing nicotine normally in response to methyl jasmonate applications, had attenuated wound induced JA and nicotine responses. Moreover, larvae consumed 1.7- to 2.7-times more leaf tissue from TMV-inoculated plants than from mock-inoculated plants. Uninoculated plants growing in chambers downwind of either TMV-inoculated plants or vials releasing MeSA at 83- to 643-times the amount TMV-inoculated plants release, exhibited normal wound-induced responses. We conclude that tobacco plants, when inoculated with TMV, are unable to elicit normal wound responses, due likely to the inhibition of JA production by the systemic increase in SA induced by virus-inoculation. The release of volatile MeSA from inoculated plants is not sufficient to influence the wound-induced responses of neighboring plants. PMID- 10467035 TI - Gravitropism of hypocotyls of wild-type and starch-deficient Arabidopsis seedlings in spaceflight studies. AB - The major purpose of this spaceflight project was to investigate the starch statolith hypothesis for gravity perception, and a secondary goal was to study plant growth and development under spaceflight conditions. This research was based on our ground studies of gravity perception in the wild type and three starch-deficient (one starchless and two reduced starch) mutants of Arabidopsis thaliana (L.) Heynh. Dark-grown seedlings that developed in microgravity were given one of several (30 min, 60 min, or 90 min) 1-g stimuli by an on-board centrifuge, and additional controls for seedling development also were performed. These latter control experiments included a morphological study of plants that developed in space in microgravity (F microg), in space on a centrifuge (F 1g), on the ground (G 1g), and on a rotating clinostat on the ground. Since elevated levels of ethylene were reported in the spacecraft atmosphere, additional controls for morphology and gravitropism with added ethylene also were performed. While exogenous ethylene reduced the absolute magnitude of the response in all four strains of Arabidopsis, this gas did not appear to change the relative graviresponsiveness among the strains. The relative response of hypocotyls of microgravity-grown seedlings to the stimuli provided by the in-flight centrifuge was: wild type > starch-deficient mutants. Although the protoplast pressure model for gravity perception cannot be excluded, these results are consistent with a statolith-based model for perception in plants. PMID- 10467036 TI - Gas-exchange analysis of chloroplastic fructose-1,6-bisphosphatase antisense potatoes at different air humidities and at elevated CO(2). AB - Gas-exchange measurements were performed to analyze the leaf conductances and assimilation rates of potato (Solanum tuberosum L. cv. Desiree) plants expressing an antisense construct against chloroplastic fructose-1,6-bisphosphatase (FBPase, EC 3.1.3.11) in response to increasing photon flux densities, different relative air humidities and elevated CO(2) concentrations. Assimilation rates (A) and transpiration rates (E) were observed during a stepwise increase of photon flux density. These experiments were carried out under atmospheric conditions and in air containing 500 micromol mol(-1) CO(2). In both gas atmospheres, two levels of relative air humidity (60-70% and 70-80%) were applied in different sets of measurements. Intercellular CO(2) concentration, leaf conductance, air-to-leaf vapour pressure deficit, and instantaneous water-use efficiency (A/E) were determined. As expected, assimilation rates of the FBPase antisense plants were significantly reduced as compared to the wild type. Saturation of assimilation rates in transgenic plants occurred at a photon flux density of 200 micromol m( 2) s(-1), whereas saturation in wild type plants was observed at 600 micromol m( 2) s(-1). Elevated ambient CO(2) levels did not effect assimilation rates of transgenic plants. At 70-80% relative humidity and atmospheric CO(2) concentration the FBPase antisense plants had significantly higher leaf conductances than wild-type plants while no difference emerged at 60-70%. These differences in leaf conductance vanished at elevated levels of ambient CO(2). Stomatal response to different relative air humidities was not affected by mesophyll photosynthetic activity. It is suggested that the regulation of stomatal opening upon changes in photon flux density is merely mediated by a signal transmitted from mesophyll cells, whereas the intercellular CO(2) concentration plays a minor role in this kind of stomatal response. The results are discussed with respect to stomatal control by environmental parameters and mesophyll photosynthesis. PMID- 10467037 TI - Subcellular localization and topology of homogalacturonan methyltransferase in suspension-cultured Nicotiana tabacum cells. AB - Pectin is a complex polysaccharide in the primary walls of all plant cells that is thought to be synthesized in the cellular endomembrane system and inserted into the wall via exocytosis. The most abundant pectic polysaccharide, homogalacturonan, is partially methylesterified within the cell by the pectin methyltransferase homogalacturonan methyltransferase (HGA-MT). The subcellular location of HGA-MT activity was determined in tobacco (Nicotiana tabacum L. cv. Samsun) cell membranes separated on linear sucrose gradients. The activity of HGA MT and two enzymatic markers of the Golgi apparatus, IDPase and UDPase, were found to be located in the same membrane fraction. No NADH cytochrome c reductase activity, a marker for the endoplasmic reticulum, was detected in the Golgi fraction. Homogalacturonan methyltransferase activity was not reduced by protease treatment of intact membranes or membranes treated with 0.01% Triton X-100. In contrast, HGA-MT activity was reduced by protease treatment of membranes permeabilized with 0.02% Triton X-100. The sensitivity of HGA-MT in detergent permeabilized membranes, and the lack of inhibition of HGA-MT activity by protease-treatment of intact membranes, provides evidence that the catalytic site of HGA-MT is located on the lumenal side of the Golgi. PMID- 10467038 TI - Characterization of auxin-binding protein 1 from tobacco: content, localization and auxin-binding activity. AB - There is evidence that auxin-binding protein 1 (ABP1) is an auxin receptor on the plasma membrane. Maize (Zea mays L.) possesses a high level of auxin-binding activity due to ABP1, but no other plant source has been shown to possess such an activity. We have analyzed the ABP1 content of tobacco (Nicotiana tabacum L.) to examine whether or not the ABP1 content of maize is exceptionally high among plants. The ABP1 content of tobacco leaves was shown by quantitative immunoblot analysis to be between 0.7 and 1.2 microg ABP1 per gram of fresh leaf. This value is comparable to the reported value in maize shoots, indicating that ABP1 is present at a similar level in both monocot and dicot plants. The ABP1 content of tobacco leaves was increased up to 20-fold by expression of a recombinant ABP1 gene, and decreased to half of the original value by expression of the antisense gene. Although ABP1 was found mainly in the endoplasmic reticulum fraction, a secreted protein showing a molecular size and epitopes similar to intracellular ABP1 was also detected in the culture medium of tobacco leaf disks. The secretion of this protein was dependent on the expression level of the ABP1 gene. PMID- 10467039 TI - Photoadaptation of two members of the Chlorophyta (Scenedesmus and Chlorella) in laboratory and outdoor cultures: changes in chlorophyll fluorescence quenching and the xanthophyll cycle. AB - The role of the xanthophyll cycle in the adaptation of two chlorococcal algae Scenedesmus quadricauda and Chlorella sorokiniana to high irradiance was studied under laboratory and outdoor conditions. We wished to elucidate whether the xanthophyll cycle plays a key role in dissipating the excesses of absorbed light, as in higher plants, and to characterise the relationship between chlorophyll fluorescence parameters and the content of xanthophyll-cycle pigments. The xanthophyll cycle was found to be operative in both species; however, its contribution to overall non-photochemical quenching (NPQ) could only be distinguished in Scenedesmus (15-20% of total NPQ). The Scenedesmus cultures showed a larger pool of xanthophyll-cycle pigments than Chlorella, and lower sensitivity to photoinhibition as judged from the reduction of maximum quantum yield of photosystem II. In general, both algae had a larger xanthophyll-cycle pool when grown outdoors than in laboratory cultures. Comparing the two species, Scenedesmus exhibited a higher capacity to adapt to high irradiance, due to an effective quenching mechanism and high photosynthetic capacity; in contrast, Chlorella represents a species with a larger antennae system, less-efficient quenching and lower photosynthetic performance. Non-photochemical quenching (NPQ) induced through the xanthophyll cycle can, to a limited extent, represent a regulatory factor in diluted algal cultures grown in outdoor solar photobioreactors, as well as in natural algal phytoplankton populations exposed transiently to high irradiance. However, it does not play an appreciable role in dense, well-mixed microalgal suspensions. PMID- 10467040 TI - Involvement of indole-3-acetic acid in the circadian growth of the first internode of Arabidopsis. AB - The extension rate of the first inflorescence node of Arabidopsis was measured during light/dark or continuous light exposure and was found to exhibit oscillations which showed a circadian rhythmicity. Decapitation induced a strong inhibition of stem extension. Subsequent application of IAA restored growth and the associated extension-rate oscillations. In addition, IAA treatments, after decapitation, re-established the circadian rhythmicity visible in the intact plants during free run. This indicates that the upper zone of the inflorescence has a major influence on the extension rate of floral stems and implies a role for auxin. Application of N-(1-naphthyl)phthalamic acid, an IAA transport inhibitor, to an intact floral stem inhibited growth and the rhythmicity in the extension rate oscillations, indicating that IAA polar transport may play a role in the dynamics of stem elongation. Furthermore, IAA-aspartate application, after decapitation, did not restore growth and rhythmicity. Nevertheless, biochemical analysis of IAA and IAA-aspartate demonstrated circadian fluctuations of the endogenous levels of both compounds. These observations suggest that IAA metabolism is an essential factor in the regulation of the circadian growth rhythm of Arabidopsis floral stems. PMID- 10467041 TI - Purification and characterisation of a novel starch synthase selective for uridine 5'-diphosphate glucose from the red alga Gracilaria tenuistipitata. AB - Red algae (Rhodophyceae) are photosynthetic eukaryotes that accumulate starch granules in the cytosol. Starch synthase activity in crude extracts of Gracilaria tenuistipitata Chang et Xia was almost 9-fold higher with UDP[U-14C]glucose than with ADP[U-14C]glucose. The activity with UDP[U-14C]glucose was sensitive to proteolytic and oxidative inhibition during extraction whilst the activity with ADP[U-14C]glucose appeared unaffected. This indicates the presence of separate starch synthases with different substrate specificities in G. tenuistipitata. The UDPglucose: starch synthase was purified and characterised. The enzyme appears to be a homotetramer with a native M(r) of 580 kDa and displays kinetic properties similar to other alpha-glucan synthases such as stimulation by citrate, product (UDP) inhibition and broad primer specificity. We propose that this enzyme is involved in cytosolic starch synthesis in red algae and thus is the first starch synthase described that utilises UDPglucose in vivo. The biochemical implications of the different compartmentalisation of starch synthesis in red algae and green algae/plants are also discussed. PMID- 10467042 TI - Manipulation of S-adenosylmethionine decarboxylase activity in potato tubers. An increase in activity leads to an increase in tuber number and a change in tuber size distribution. AB - S-Adenosylmethionine decarboxylase (SAMDC; EC 4.1.1.50) is a key enzyme in the biosynthesis of the polyamines spermidine and spermine from putrescine and its activity is rate limiting in this pathway. Transgenic potato (Solanum tuberosum L. cv. Desiree) plants containing both sense and antisense SAMDC constructs driven by the tuber-specific patatin promoter have been generated and analysed. In sense transformants, developing tubers expressed higher steady-state levels of the SAMDC-specific transcript, had higher levels of SAMDC activity and contained significantly higher levels of spermidine than vector-transformed controls. Additionally, there was a significant shift in tuber size distribution with larger numbers of smaller tubers but no overall change in tuber yield. In developing tubers from the antisense transformed lines, there was a decrease in SAMDC transcript level, SAMDC activity and total polyamine levels. However, no obvious phenotypic effect was detected in the tuberisation physiology of the antisense lines. PMID- 10467043 TI - Validation and characterization of the computerized laryngeal analyzer (CLA) technique. AB - The aim of this study was to investigate the response characteristics of the Computerized Laryngeal Analyzer (CLA) and the validity of the noninvasive CLA method to detect swallowing-induced laryngeal elevation correctly. Two healthy adults and two experimental models were used in the study. The CLA technique identified all swallowing events but was unable to discriminate between swallowing and other movements of the tongue or the neck. The computer program produced a derivated response to a square wave signal. Stepwise bending increments of the sensor displayed a linear amplitude response. The degree of laryngeal elevation could not be estimated with the CLA technique, and it was not possible to draw any reliable conclusions from the recordings as to whether the larynx was moving upward or downward. PMID- 10467044 TI - Vagal stimulation for reciprocal coupling between glottic and upper esophageal sphincter activities in the canine. AB - Glottic adduction couples with relaxation of the tonically contracted upper esophageal sphincter (UES) to constitute the end point of pharyngeal swallowing. Together with deglutitive laryngeal elevation, this reciprocal relationship contributes to protecting the lungs from aspiration. Degrees of uncoordination between glottic and upper esophageal sphincters can be seen under diverse circumstances of neurologic damage such as stroke, gastroesophageal reflux, and in the growing elderly population presenting with weaker musculature. We hypothesized that reciprocal coupling between glottic closure and UES relaxation may be artificially reestablished through vagal stimulation if the appropriate neural centers and their leading pathways remain capable of exciting a critical number of motor units. Orderly recruitment of the vagus nerve was produced in three dogs with a circuit superimposing 600-Hz, 3800-0-microA blocking over 10-70 Hz, 0-2300-microA stimulating currents. Amplitudes of motor unit or compound muscle action potentials were recorded from the thyroarytenoideus and cricopharyngeus via surface electromyographic electrodes. Stimulation was accompanied by a drop in UES intraluminal pressure. The reciprocal relationship between the two muscles could be time locked within certain stimulus parameters. We submit that electronic coupling between the glottic and UESs may show promise in preventing aspiration under selected circumstances. PMID- 10467045 TI - Arterial oxygen saturation in severely disabled people: effect of oral feeding in the sitting position. AB - Control of the circulatory and respiratory systems is especially important in severely disabled people. The purpose of this study was to clarify the response of hemoglobin oxygen saturation level (SpO(2)), pulse rate, and respiratory rate during oral feeding in severely disabled persons. Continuous measurement of these variables was done by pulse oximetry and respiratory inductance plethysmography under two experimental settings in eight severely disabled persons aged 14-28 yrs. Setting I consisted of the following three procedures: (a) a 30-min period in the supine position, (b) a 50-min period in a sitting position, and (c) a 30 min period in the supine position. Setting II consisted of the following four procedures: (a) a 30-min period before the meal in the supine position, (b) a nonspecified period in a sitting position during which the meal was taken, (c) a 30-min period after the meal in the same sitting position, and (d) a 30-min period in the supine position. Results showed that mean SpO(2) level decreased and mean pulse rate increased during the meal in almost all subjects. In many cases, pulse rate and SpO(2) level did not return to baseline values in the sitting position after the meal. These findings indicate that oral feeding of severely disabled persons in a sitting position places considerable stress on the circulatory system, the effects of which may last after the meal in some cases. PMID- 10467046 TI - Swallowing abnormalities after acute stroke: A case control study. AB - Dysphagia is a common and potentially fatal complication of acute stroke. However, the underlying pathophysiology, especially the relative importance of motor and sensory dysfunction, remains controversial. We conducted a case control study of 23 acute stroke patients (mean age = 72 yr) at a median of 6 days post stroke and 15 healthy controls (mean age = 76 yr). We used novel methods to assess swallowing in detail, including a timed videoendoscopic swallow study and oral sensory threshold testing using electrical stimulation. Vocal cord mobility and voluntary pharyngeal motor activity were impaired in the stroke group compared with the controls (p = 0.01 and 0.03). There was a delay during swallowing in the time to onset of epliglottic tilt in the stroke group, particularly for semisolids (p = 0.02) and solids (p = 0.01), consistent with a delay in initiation of the swallow. Sensory thresholds were not increased in the stroke group compared with controls. We conclude that pharyngeal motor dysfunction and a delay in swallow initiation are common after acute stroke. Vocal cord mobility is reduced, and this may result in reduced airway protection. We found no evidence to support the hypothesis that oropharyngeal sensory dysfunction is common after acute stroke. PMID- 10467047 TI - Contour tracking using a knowledge-based snake algorithm to construct three dimensional pharyngeal bolus movement. AB - Videofluorography (VFG) using a barium-mixed bolus is in wide clinical use for assessing patients with swallowing disorders. VFG is usually done with both lateral (LA) and anterior-posterior (AP) views, most commonly in two separate sittings. A real-time, three-dimensional (3-D) representation of the evolution of a pharyngeal bolus and its volumetric information can potentially help clinicians analyze and visualize the kinematics of swallowing, dysphagia, and compensatory therapeutic strategies. Active contour models, also known as "Snakes," have been used to solve various image analysis and computer vision problems. We applied a Snake algorithm to automate in part the contour tracking and reconstruction of VFG images to visualize and quantitatively analyze the 3-D evolution of a pharyngeal bolus. To improve the accuracy of the Snake search, we provided the additional "knowledge" of the pharyngeal image itself, which served as an extra constraint to push the Snake curve toward the desired contour. VFG of pharyngeal bolus transport in a normal subject was recorded by using barium-mixed boluses (viscosity: 185 centipoise, density: 2.84 g/cc) with volumes of 5, 10, and 20 ml. The resulting LA and AP video images were digitally captured and matched frame by frame. The knowledge-based Snake search algorithm was used to generate Snake points to satisfy both internal (i.e., smoothness) and external (i.e., boundary fitting) constraints. Using these Snake points, we traced the 3-D bolus movement at each time instant, assuming elliptic geometry in the cross-section of the pharyngeal bolus. By concentrating the 3-D images for each time instant, we developed a 3-D movie representing pharyngeal bolus movement. The efficiency, reproducibility, and accuracy of this algorithm in tracing pharyngeal bolus boundaries and estimating front/tail velocities were assessed and found satisfactory. We conclude that 3-D pharyngeal bolus movement can be traced both accurately and efficiently by using a knowledge-based Snake search algorithm. PMID- 10467048 TI - Differentiation of normal and abnormal airway protection during swallowing using the penetration-aspiration scale. AB - Accidental loss of food or liquids into the airway while eating or drinking is perhaps the most clinically significant consequence of dysphagia. Although videofluoroscopic recording of swallowing is the current gold standard for identifying and determining remediation for aspiration, results are generally described in descriptive terms, thus limiting information and lending to errors of interpretation. We previously published an 8-point scale to quantitate selected aspects of penetration and aspiration conveying depth of airway invasion and whether or not material entering the airway is expelled (Rosenbek et al., 1996, Dysphagia 11:93-98). The present study defines the distribution of the Penetration-Aspiration Scale scores in healthy normal subjects of different genders and ages. The scale was also used with two groups of patients known to have significant dysphagia relative to stroke or head and neck cancer. Significant differences found among groups are discussed. PMID- 10467050 TI - Comments on selected recent dysphagia literature PMID- 10467049 TI - Do we know what is normal and abnormal airway protection? PMID- 10467051 TI - Selected abstracts from the 14th congress of the international association of dentistry for the handicapped and the 15th annual meeting of the japanese society of dentistry for the handicapped, yokohama, japan, september 3-6, 1998 PMID- 10467052 TI - Protease interactions with bacillus thuringiensis insecticidal toxins AB - The microbe Bacillus thuringiensis (Bt) produces crystals that contain insecticidal crystal proteins (ICPs) used to control many major pests. ICPs are degraded by proteases from a variety of sources, including those endogenous to the bacterium, those purified from animals and plants, or those found in insects. Proteases in the bacterium function in protein metabolism during sporulation; in some cases they hydrolyze ICPs. Insect proteases are implicated in Bt toxin specificity, mode of action and insect adaptation to Bt. This review describes the current knowledge of protease interactions with ICPs with special emphasis on the role of proteases in insect resistance to Bt toxins. Arch. Copyright 1999 Wiley-Liss, Inc. This article is a US Government work and, as such, is in the public domain in the United States of America. PMID- 10467053 TI - Phytosterol metabolism and absorption in the generalist grasshopper, Schistocerca americana (Orthoptera: Acrididae). AB - A series of experiments, using GLC, RP-HPLC, and GC-MS techniques, were performed to examine the metabolic fate and absorption of different dietary sterols in the grasshopper Schistocerca americana. In the first experiment, grasshoppers were reared on diets containing different sterols presented singly. Cholesterol was the dominant tissue sterol recovered from cholesterol and "soybean sitosterol" fed grasshoppers but among the grasshoppers fed diets with stigmasterol and spinach sterols (both unsuitable for growth and development), the amount of cholesterol recovered was not different from that of newly hatched grasshoppers. In the second experiment, grasshoppers were given diets containing mixtures of soybean sitosterol and stigmasterol and the metabolic fate of these dietary sterols was recorded. Results from this experiment suggest that the presence of an unsuitable dietary sterol does not interfere with cholesterol production from sitosterol. They also demonstrate that large quantities of unmetabolized dietary sterols with C-24 ethyl groups are recovered from grasshoppers fed diets containing stigmasterol. Finally, tissue sterol profiles of grasshoppers with and without their midguts were compared. Results suggest that the midgut is the major tissue where unmetabolized dietary sterols accumulate. How these sterol metabolic constraints impact development and survival is discussed as well as the impact they might have on grasshopper feeding behavior. Arch. PMID- 10467054 TI - Ascorbic and uric acid responses to xanthotoxin ingestion in a generalist and a specialist caterpillar AB - For herbivorous insects, dietary sources of low molecular weight non-enzymatic antioxidants, such as ascorbic acid, may influence performance in the presence of phototoxic plant constituents. We examined responses of Trichoplusia ni, a broad generalist, and Depressaria pastinacella, a specialist on furanocoumarin containing plants, to variation in dietary ascorbic acid availability in the presence and absence of xanthotoxin, a phototoxic furanocoumarin. In T. ni, dietary ascorbic acid significantly increased levels of this compound in body tissues (approximately 7-fold, 5-fold, and 8-fold in hemolymph, gut, and fat body, respectively). In the presence of xanthotoxin, however, the amount of ascorbic acid accumulated significantly decreased. This decrease was not due to antifeedant effects of xanthotoxin and may instead have resulted from depletion of ascorbic acid due to its radical scavenging activity. In contrast, ascorbic acid levels in D. pastinacella were less affected by variation in dietary levels of either xanthotoxin or ascorbic acid, although uric acid, another potential water-soluble nonenzymatic antioxidant, increased in response to dietary ascorbic acid, as it did in T. ni. Thus, for generalists, such as T. ni, that lack specialized detoxification mechanisms against phototoxins such as furanocoumarins, dietary ascorbic acid may play an important role in antioxidant defense, and, for caterpillars in general, uric acid may also contribute to antioxidant defenses. Arch. Copyright 1999 Wiley-Liss, Inc. PMID- 10467055 TI - Effects of total dietary nitrogen and nitrogen form on the development of xylophagous leafhoppers AB - The influence of primary nutrients on the development of the leafhopper Homalodisca coagulata (Say) was examined using Glycine max (L.) Merrill subjected to treatments using different forms of nitrogen fertilization (urea and nitrate) and inoculation with Rhizobium bacteria. Concentrations of amino acids, organic acids, carbohydrates, and ureides in xylem fluid varied with treatment; differences were most pronounced in young plants (6 weeks after planting), and subsided as plants aged. Plants receiving urea fertilization had lower concentrations of organic nitrogen and carbon, but more balanced profiles of amino acids. Leafhoppers on urea-fertilized plants had much higher success in development (62 to 87% survival to adult stage) than H. coagulata developing on nitrate-fertilized plants (31-42%). The sex ratio of successfully developed adults varied greatly with treatments, indicating that female survival was much higher on urea-fertilized plants. Males and females utlilized diets differently as indicated by the carbon and nitrogen content of adult carcasses. Consumption rates and nitrogen assimilation efficiencies were higher for young (third instar) leafhoppers on urea-fertilized plants. Instar duration and total time of development were not affected by treatment. Highest rates of success in development on diets providing the lowest (yet most balanced) concentrations of total nitrogen, amino acids, and organic compounds underscored the importance of nutrient balance for leafhopper development. Arch. Copyright 1999 Wiley-Liss, Inc. PMID- 10467056 TI - Resistance to bacillus thuringiensis Cry1Ac toxin in three strains of heliothis virescens: proteolytic and SEM study of the larval midgut AB - In a previous study, we demonstrated that resistance to Bacillus thuringiensis toxins in Heliothis virescens might be related to differences in the composition of the proteolytic extracts from insect midgut. There, we found specific proteolytic bands present in the gut extracts of the resistant strain and absent from the susceptible one. Here we report related facts using a new resistant strain (KCB) and a cross between the two strains used in our previous study. As would be expected, no quantitative differences in total proteolytic activity were found between the strains, although qualitative differences related to the presence or absence of specific proteolytic activity bands using SDS-PAGE could be responsible for the observed resistance phenomenon. Moreover, an SEM study made at different time intervals after intoxication shows that in the initial hours following intoxication, both the susceptible and the resistant strains show significant damage to the midgut epithelium. In the interval between 3 and 48 h, however, the resistant strain recovered such that by 48 h it had fully recovered whereas the susceptible strain did not. Arch. Copyright 1999 Wiley-Liss, Inc. PMID- 10467057 TI - Feeding preferences of spodoptera exigua in response to form and concentration of selenium AB - Minimal information is available on the impact of various organic and inorganic forms of the ecologically and agriculturally important pollutant, selenium (Se), on insect herbivores. We conducted bioassays with artificial diet to examine the feeding responses of a generalist herbivore, Spodoptera exigua (Hubner) (Lepidoptera: Noctuidae), to various forms and concentrations of Se. Two different-aged cohorts of larvae were examined in choice tests with control diets vs. test diets incorporating lethal concentrations (LC(10), LC(30), LC(50), and LC(70)) of sodium selenate, sodium selenite, seleno-DL-cystine, and seleno-DL methionine. Tests initiated with neonates showed larvae significantly preferred control diet over diet with sodium selenate, sodium selenite, or selenocystine, but at most concentrations showed no preference between selenomethionine and control diet. Choice tests initiated with third instars demonstrated a preference for control diet over sodium selenate treatments, and sodium selenite treatments. In contrast, no significant responses were found in tests initiated with third instars offered the choice between selenocystine or selenomethionine and untreated controls. Additionally, comparisons of consumption demonstrated that inorganic selenium compounds were antifeedants whereas the organic selenium compounds tested have little antifeedant activity. The toxicity of all of the tested forms of selenium, in combination with the lack of antifeedant activity of some compounds, has the potential to affect both the distribution and diversity of terrestrial herbivores in both agricultural and natural systems. Arch. Copyright 1999 Wiley-Liss, Inc. PMID- 10467058 TI - Feeding rate and the structure of protein digestion and absorption in lepidopteran midguts AB - The transformation of leaf protein into insect tissue can be distilled grossly into four serial steps: consumption of protein, digestion of protein into small peptides and free amino acids, absorption of these fragments across the gut epithelium, and construction of tissue from the absorbed fragments. We examine these steps in caterpillars, with emphasis on determining which, if any, process is limiting and how digestive events influence-and are influenced by-the rate of consumption. Five competing hypotheses are (1) the steps are matched, (2) consumption is limiting, (3) digestion is limiting, (4) absorption is limiting, and (5) post-absorptive processes are limiting. For conditions appropriate to each of the five hypotheses, we use a chemical reactor model to predict the concentration profiles of protein and protein breakdown products along the midgut. We test the predictions in two ways. First, based on midgut morphology and rates of digesta flow and protein breakdown estimated from in vitro experiments, we calculate the axial concentration profile of protein in the midgut of fifth instar Manduca sexta. This calculation suggests that protein breakdown occurs primarily in the anterior midgut, thus excluding the hypotheses that feeding and digestive processes are matched or that digestion is limiting. In addition, caterpillar feeding responses to reduced concentrations of protein are not consistent with the hypothesis that consumption is limiting. Second, we review published studies that show midgut concentration profiles of protein and breakdown products, one supporting the hypothesis that absorption across the midgut epithelium is limiting and the other suggesting that post-absorptive processes are limiting. Consequently, we interpret compensatory feeding by larval M. sexta as a mechanism for maintaining saturated rates of protein absorption or of tissue construction. Arch. Copyright 1999 Wiley-Liss, Inc. PMID- 10467059 TI - Protein hydrolysis by colorado potato beetle, leptinotarsa decemlineata, digestive proteases: the catalytic role of cathepsin D AB - Although several studies were carried out over the last 15 years to assess the nature and characteristics of digestive proteases in herbivorous insects, little is known about the relative importance of these enzymes in the hydrolysis of specific dietary proteins. In this study, we assessed the involvement of Colorado potato beetle (CPB; Leptinotarsa decemlineata Say, Chrysomelidae) aspartate, cysteine, and serine digestive proteinases in the degradation of two model substrates: ribulose biphosphate carboxylase/oxygenase, the major protein in potato leaves, and the pro-region of papaya proteinase IV, a cysteine protease inhibitor (PI) susceptible to proteolysis by the insect "nontarget" proteases. As shown by the use of various combinations of diagnostic PIs specific to the different classes of CPB proteinases, the insect aspartate (cathepsin D-like) proteinase activity is important in initiating the hydrolysis of both proteins when the insect is feeding on potato, while cysteine (cathepsin B/cathepsin H like) and serine (chymotrypsin-like) proteinase activities would be involved in subsequent steps of the hydrolytic process. Similar observations were made with diet-induced variants of the insect protease system, suggesting the importance of digestive cathepsin D and the sequential hydrolysis of dietary proteins in CPB, regardless of the diet ingested. Based on these observations, a preliminary model is proposed to explain dietary protein hydrolysis in CPB, also taking into account the information currently available about the distribution of digestive endo- and exopeptidases in the midgut of CPB. The potential of a wound-induced cathepsin D inhibitor from tomato in developing CPB-resistant transgenic potato lines is also discussed, after demonstrating the "pepstatin-like" effect of a recombinant form of this proteinaceous inhibitor against the insect cathepsin D. Arch. Copyright 1999 Wiley-Liss, Inc. PMID- 10467060 TI - Salivary glucose oxidase: multifunctional roles for helicoverpa zea? AB - Labial glands of Helicoverpa zea produced a glucose oxidase (GOX), which was present in the saliva and midgut lumen. We purified GOX 23-fold by isoelectric focusing of labial gland homogenates and investigated physical and kinetic properties of the enzyme. D-glucose and 6-deoxy-D-glucose were the optimal substrates of 22 carbohydrates tested with GOX. The enzyme was not inhibited by several inhibitors of fungal GOX but was sensitive to HgCl(2). Labial gland GOX activities varied daily during larval development with highest activities found when larvae were actively feeding. Based on pH optimum, pI, molecular weight estimate and K(m(glucose)), the insect enzyme is not derived from fungal GOXs but appears to have similar kinetic and physical attributes to other insect GOXs. Some possible functions are discussed, including antimicrobial properties, manipulating midgut O(2) levels, altering host plant defense responses, and metabolizing carbohydrates. Arch. Copyright 1999 Wiley-Liss, Inc. PMID- 10467061 TI - Preface PMID- 10467062 TI - Intracoronary flow velocity measurements in adjacent stenotic and normal coronary arteries during incremental intravenous dobutamine stress and intracoronary adenosine injection. AB - To investigate the concomitant coronary flow and hemodynamic changes induced by dobutamine and adenosine in the catheterization laboratory, we studied stenotic and adjacent normal coronary arteries in 20 patients using paired Doppler Flowires. Coronary flow velocity and hemodynamics were measured sequentially after intracoronary (ic) adenosine, during incremental iv dobutamine infusion, and after the addition of ic adenosine during sustained peak dobutamine stress (adenosine on dobutamine). Distal to stenotic arteries, average peak velocity (APV) increased significantly (from 11 +/- 5 to 16 +/- 7 cm/sec, P < 0.001) at an intermediate dose of dobutamine (20 microg/kg/min, Dobutamine20) but did not change further thereafter to peak dobutamine stress (17 +/- 7 cm/sec), despite the significant further increase in rate-pressure product (RPP). Peak stress APV did not change with adenosine on dobutamine (to 18 +/- 7 cm/sec). In normal arteries, APV increased at Dobutamine20 (from 20 +/- 7.5 to 30 +/- 12 cm/sec, P < 0.01) and further at peak dobutamine stress (to 42 +/- 10 cm/sec, P < 0.0001) always exceeding the concomitant significant increases in RPP. Peak stress APV increased further with adenosine on dobutamine (to 53 +/- 13 cm/sec, P < 0.001). Our data demonstrate that at peak dobutamine stress there is supply/demand mismatch only in stenotic arteries where coronary flow reserve is exhausted at an intermediate dobutamine dose. Furthermore, adenosine on dobutamine potentiates coronary flow heterogeneity between stenotic and normal adjacent arteries. Cathet. Cardiovasc. Intervent. 48:1-9, 1999. PMID- 10467063 TI - Coronary flow heterogeneity and dobutamine-induced myocardial stress testing: value of measurements in stenotic and adjacent normal arteries. PMID- 10467064 TI - Coronary and peripheral artery remodeling in patients undergoing PTCA: An intracoronary and transcutaneous ultrasound study. AB - The aim of this study was to assess the relationship between the plaque burden of nonstenotic coronary artery segments and the wall thickness of peripheral arteries using intracoronary and transcutaneous ultrasound imaging, respectively. Intracoronary ultrasound (CVIS, 3.5 Fr) was performed in 27 patients undergoing percutaneous transluminal coronary angioplasty. Carotid arteries were imaged by B mode ultrasound with semiautomatic edge detection and radial arteries by high resolution A-mode echotracking (NIUS 2). Quantitative measurements included coronary artery intima-media cross-sectional area (IM(CSA)) and cross-sectional narrowing (CSN), as well as intima-media thickness (IMT) and lumen radius (r) of the common carotid and the radial arteries. Intima-media thickness was increased in coronary, carotid, and radial arteries. Coronary arteries had an IM(CSA) of 7.7 +/- 2.5 mm(2) and a CSN of 24% +/- 8%. Despite this moderate plaque burden, lumen area was preserved (12.3 +/- 4.2 mm(2)) because of compensatory enlargement of coronary arteries. Right and left carotid and right radial arteries had an IMT of 575 +/- 78 microm, 570 +/- 129 microm, and 328 +/- 61 microm, respectively. There was no correlation between coronary IM(CSA) and carotid IMT (r = 0.07) or radial IMT (r = 0.02), and there was no correlation between coronary CSN and carotid IMT/r (r = 0.12), or radial IMT/r (r = 0.25). In conclusion, in these patients with symptomatic ischemic disease no relationship between IMT of the coronary arteries and IMT of carotid or radial arteries was found. Although increasingly popular, IMT of peripheral arteries may be of limited value as surrogate marker for the severity of coronary artery disease. Cathet. Cardiovasc. Intervent. 48:12-17, 1999. PMID- 10467065 TI - Accuracy of quantitative ventriculographic analysis at reduced frame rates. AB - OBJECTIVE: We measured the error in left ventricular (LV) volume and wall motion measurement from recording contrast ventriculograms at 15 frames/sec instead of 30 frames/sec. BACKGROUND: Angiograms are increasingly recorded at 15 frames/sec instead of 30 frames/sec to improve efficiency. However the resulting error in quantitative analysis has not been evaluated. METHODS: Contrast ventriculograms recorded at 30 frames/sec of 23 patients undergoing diagnostic studies were digitized from cine film. End diastolic volume (EDV), end systolic volume (ESV), ejection fraction, anterior wall motion, and inferior wall motion were measured from a full set recorded at 30 frames/sec and from two reduced data sets at 15 frames/sec generated from the even and odd numbered frames. Intraobserver variability was compared with the error due to frame rate reduction. RESULTS: The error due to frame rate reduction did not differ significantly from intraobserver variability (p > 0.1 for all), measuring 2.6 +/- 2.2 ml (r = 0.997) vs. 1.9 +/- 1.2 ml (r = 0.999), respectively for EDV, 2.4 +/- 1.8 ml (r = 0.991) vs. 2.1 +/- 1.7 ml (r = 0.994), respectively for ESV, and 0.2 +/- 0.2 percent (r = 0. 981) vs. 0.1 +/- 1.2 percent (r = 0.994), respectively for ejection fraction. The error and variability for wall motion were 0.3 +/- 0.3 SD (r = 0.928) vs. 0.2 +/- 0.1 SD (r = 0.946), respectively for the anterior wall and 0.2 +/- 0.2 SD (r = 0.963) vs. 0.2 +/- 0.1 SD (r = 0.968), respectively for the inferior wall. CONCLUSION: Imaging the LV at 15 frames/sec does not cause a significant error in measuring LV volume and function compared with imaging at 30 frames/sec. Cathet. Cardiovasc, Intervent. 48:18-21, 1999. PMID- 10467066 TI - Angiography at reduced frame rates: how low can you go? PMID- 10467067 TI - Focal wall overstretching after high-pressure coronary stent implantation does not influence restenosis. AB - To determine if vessel wall overstretching during coronary stenting is associated with a higher restenosis rate, the intravascular ultrasound morphological evaluation was performed following ultrasound criteria. A total of 468 lesions with successful coronary Palmaz-Schatz stenting guided by intravascular ultrasound were classified into the no overstretching group (n = 295) and the overstretching group (n = 147). There were 26 lesions not classifiable due to the poststent morphology. Balloon-to-vessel ratio was 1.12 +/- 0.17 in the no focal overstretching group and 1. 20 +/- 0.20 in the overstretching group (P < 0.0002). Follow-up angiogram was performed in 77% of no focal overstretching group and in 75% of the focal overstretching group. The restenosis rate of the no focal overstretching group was 19.8% and 20.9% in the focal overstretching group, respectively (P = 0.65). Focal overstretching was more frequent following balloon oversizing. No increase in restenosis rate, found in focal overstretched stented lesions, leads us to the hypothesis of a regulation of smooth-muscle-cell proliferation activated by the normalization of blood flow and of shear stress, when stent implantation succeeds in optimally improving the lumen. Cathet. Cardiovasc. Intervent. 48:24-30, 1999. PMID- 10467068 TI - Coronary artery aneurysms: A 25-patient study. AB - Coronary artery aneurysm is a rare disorder, characterized by abnormal dilatation of a localized portion or diffuse segments of the coronary artery. We studied clinical demographics, catheterization findings, and clinical outcomes in an Asian patient cohort with documented coronary artery aneurysms. Compared to a Caucasian adult population, our patient cohort had a lower incidence of coronary artery aneurysm (0.25% vs. 2.6%), and more patients with nonobstructive coronary artery aneurysms (70%); age, gender, and coronary distribution were comparable. The initial presentation of myocardial infarction occurred in five patients (5/17, 30%) with nonobstructive coronary artery aneurysms; however, none who were receiving preventive medications consisting of anticoagulant and antiplatelet agents subsequently developed myocardial infarction. We conclude that the incidence of coronary artery aneurysms with or without associated significant coronary stenosis seems to be lower in the Asian population. In contrast, the incidence of nonobstructive coronary artery aneurysms is considerably high and should not be thought of as a relatively benign disease entity if not treated with preventive medications. Rheumatoid arthritis-related vasculitis might be a cause of coronary artery aneurysm. Surgical intervention is based on the severity of coronary artery stenosis. The result of medical treatment has been compatible with long-term survival. Cathet. Cardiovasc. Intervent. 48:31-38, 1999. PMID- 10467069 TI - Minimally invasive evaluation of coronary stents with electron beam computed tomography: In vivo and in vitro experience. AB - We sought to describe the value of electron beam computed tomography (EBCT) to detect stent restenoses at follow-up and to identify characteristic EBCT features of coronary stents. Six coronary stents (GRII, Jostent, NIR, PS, Micro, Wiktor) were scanned in vitro (10 1. 5-mm-thick slices, 15-cm FoV, 100-ms AT) before and after inflation with 3.0-, 3.5-, and 4.0-mm balloons to study intensity values and different stent dimensions in comparison to caliper measurements. In 44 patients (60 +/- 10 years, 4 females) we prospectively compared EBCT findings [eight 8-mm-thick slices in the flow mode, 26-cm field of view (FoV), 50 ms acquisition time (AT)] with 6-month angiographic outcome after placement of 86 stents in 49 vessels. For the detection of significant angiographic luminal narrowing (>/=75%) that occurred in 17/49 vessels (35%), we found a sensitivity of 65%, a specificity of 84%, and a positive and negative predictive value of 69% and 82%, respectively. The in vitro data show significant differences in image intensity between the stents and a significant gradual decrease in image intensity with increasing stent diameters. The mean differences between EBCT and caliper measurements for the length and diameter were 0.17 +/- 0.18 mm and -0.32 +/- 0.25mm, respectively. EBCT is currently the only noninvasive technique that allows the assessment of stent dimensions and stent geometry, which may prove useful in patients when intravascular ultrasound cannot be performed. Contrast enhanced EBCT is a promising tool for minimally invasive stent patency evaluation at follow-up, especially in patients that are unable to exercise. Cathet. Cardiovasc. Intervent. 48:39-47, 1999. PMID- 10467070 TI - Novel approach to rotational atherectomy results in low restenosis rates in long, calcified lesions: long-term results of the San Antonio Rotablator Study (SARS). AB - Ablation technique and adjunctive strategy may affect restenosis after rotational atherectomy. To minimize trauma to the vascular wall, we changed the technique of rotablation as follows: the RPM range was decreased to 140,000-160,000 RPM, the ablation was performed using a repetitive pecking motion, avoiding a decrease in the rotational speed of the burr greater than 3,000 RPM, long lesions were divided into segments and each segment was separately ablated, and the burr-to artery ratio was intended to be approximately 0.75. To prevent coronary spasm, before and after each pass, 100-200 microg nitroglycerin and 100-200 microg verapamil i.c. boluses were administered. Adjunctive PTCA was performed using a closely sized 1.1:1 balloon-to-artery ratio with a noncompliant balloon at low pressures for 120 sec. The study incorporated 111 patients with a combined total of 146 calcified lesions. Results. A total of 31.5% of patients underwent a multivessel procedure. No deaths occurred. Q-wave MI and/or creatine kinase elevation greater than three times baseline levels occurred in 4.5% of patients. By quantitative coronary angiography (QCA), the reference vessel diameter was 3.13+/-0.59 mm, mean lesion length was 33.41+/-18.58 mm. Percent stenosis and mean luminal diameter were as follows: at baseline 75.7%+/-10.8%, or 0.76+/ 0.41mm, Post-rotational atherectomy 41.5%+/-3.6%, or 1.83+/-0.43 mm, Post-PTCA 18.2%+/-11.9%, or 2.56+/-0.50 mm. Six-month angiographic follow-up was available in 64 (57.7%) pts. Net luminal gain was 1.15+/-0.76 mm, with a late luminal loss of 0.65+/-0.84 mm. The mean diameter stenosis at follow-up was 37.6%+/-28.5%, with MLD 1.91+/-1.21 mm. The binary restenosis rate was 28.1%. Therefore, modification of rotational atherectomy technique with adjunctive PTCA resulted in a favorable restenosis rate in long, calcified lesions. Cathet. Cardiovasc. Intervent. 48:48-53, 1999. PMID- 10467072 TI - Significant pulmonary valve incompetence following oversize balloon pulmonary valveplasty in small infants: A long-term follow-up study. AB - Oversized dilatation balloons are recommended for relief of valvar pulmonic stenosis in infants and children during cardiac catheterization. Little information exists about the long-term outcome of this practice. Six of 107 consecutive patients undergoing balloon pulmonary valveplasty developed increasing pulmonary valve incompetence during follow-up periods of 0.5-10 years (mean, 7.2 years). Secondary right ventricular dilatation prompted insertion of a bioprosthetic pulmonary valve in one patient 6.8 years after intervention; valve replacement is pending in two additional patients, 4.3 and 10 years after intervention, respectively; and the three remaining subjects are thought likely to require valve replacement during childhood. The six reported subjects differ from the pulmonary valveplasty group as a whole in that they are younger (median age, 3 days vs. 11 months), had a higher degree of obstruction (right/left ventricular systolic pressure ratio prior to valveplasty 1.28 vs. 0.92), and underwent dilatation with relatively oversized balloons (balloon diameter to pulmonary valve annulus ratio 1.44 vs. 1.08). The balloon diameter to valve annulus ratio did not exceed 1.5 in any subject. Caution is advised in the use of oversized dilatation balloons in neonates or young infants with severe or critical pulmonic stenosis. Long-term consequences of substantial pulmonary valve incompetence outweigh, in our opinion, the limited gradient relief achieved with smaller balloons, sometimes requiring a second dilatation of the valve at an older age and larger size. Cathet. Cardiovasc. Intervent. 48:61-65, 1999. PMID- 10467071 TI - Reuse of catheters for percutaneous transluminal coronary angioplasty: effects on procedure time and clinical outcomes. AB - In July 1996, because of concern regarding the possible transmission of Creutzfeldt-Jakob disease, the province of Quebec stopped the reuse of percutaneous transluminal coronary angioplasty (PTCA) catheters. Prior to this time, PTCA balloon catheters were used a maximum of four times and guide catheters a maximum of two times in the cardiac catheterization laboratory at our institution. After this time, only new catheters were used. In order to examine the effects of catheter reuse on duration of PTCA procedures and clinical outcomes, we compared 53 consecutive patients undergoing PTCA prior to 21 July 1996 with 54 consecutive patients undergoing PTCA after that time. A total of 81 men and 26 women underwent PTCA (average age, 64 +/- 12 years). There were no significant differences between the single-use and reuse groups with respect to baseline characteristics. There were also no significant differences in the numbers of PTCA catheters used (97 vs. 103, P = NS) or angiographic success rates (88% vs. 83%, P = NS). There was a trend for total procedure time and fluoroscopy time to be slightly longer for single-use compared with reuse cases (49.2 vs. 45.7 min and 19.7 vs. 16.8 min, respectively; P = NS for both comparisons). However, after controlling for case severity and the use of stents, there were no significant differences in total procedure time or fluoroscopy time between the two groups. We found little evidence to suggest that the reuse of PTCA catheters is associated with longer total procedure time or fluoroscopy time. We conclude that if catheter reuse is not found to be associated with infectious disease transmission, its widespread use should be considered. Cathet. Cardiovasc. Intervent. 48:54-60, 1999. PMID- 10467073 TI - Larger balloons/annulus ratio: probably not the major problem PMID- 10467075 TI - Possible anaphylactic reaction to abciximab. AB - We report a patient who experienced a severe anaphylactic reaction during coronary artery stenting. Subsequent to administration of a weight-adjusted dose of abciximab, the patient developed profound hypotension and severe, acute airway obstruction. The reaction was successfully reversed with a 16-hr infusion of epinephrine, steroids, and an H1 blocker. Cathet. Cardiovasc. Intervent. 48:71 73, 1999. PMID- 10467074 TI - Sudden death and cerebral anoxia in a young woman with congenital ostial stenosis of the left main coronary artery. AB - We report a 36-year-old woman with ventricular fibrillation, subsequent sudden clinical cardiac death, and a prolonged brain anoxia. After a successful resuscitation coronary angiography revealed congenital ostial left main coronary artery (LMCA) stenosis. Surgical anastomosis of the left internal mammary artery (LIMA) to LAD led to a complete recovery. Postoperative electrophysiological examination, mainly programmed ventricular stimulation, failed to excite any rhythm disturbances. Cathet. Cardiovasc. Intervent. 48:67-70, 1999. PMID- 10467076 TI - Balloon rupture during stent implantation: A novel technique of salvage with a new manual power injector. AB - Pinhole leak or rupture of a stent delivery balloon is a well-recognized technical problem encountered in vascular interventions. This event leads to inadequate stent expansion. These stents cannot be fully deployed with the same balloon and frequently the balloon cannot be retrieved without dislodging the stent. We describe a technique for successful stent deployment in such situations using the Oz Power Syringe, a new manual power injector. Cathet. Cardiovasc. Intervent. 48:74-77, 1999. PMID- 10467077 TI - Percutaneous closure of a left ventricular perforation post balloon mitral valvotomy. AB - The risk of left ventricular perforation is a rare but well-recognized complication of percutaneous double balloon mitral valvuloplasty that usually requires surgical bailout. We describe a case of left ventricular perforation with cardiac tamponade, caused by the propulsion of the balloons during balloon mitral valvotomy on an 86-year-old female with previous thoracotomies that was resolved using percutaneous coil embolization of the perforation. This approach to these types of complications, although unlikely to be of extensive use, will serve to expand the horizon of options in the field of interventions. Cathet. Cardiovasc. Intervent. 48:78-83, 1999. PMID- 10467078 TI - Effects of local heparin administration on coronary thrombin activity during percutaneous transluminal coronary angioplasty. AB - Simultaneously obtained blood samples from the coronary sinus and systemic arterial circulation were analyzed for antithrombin III (ATIII) activity and fibrinopeptide A (FpA) concentration in nine patients undergoing elective PTCA in order to determine the effects of locally delivered heparin. Samples were obtained at the following designated times: prior to the administration of systemic heparin (period I); 5 min following a loading dose of systemic heparin (period II); 5 min following the final balloon inflation but prior to local delivery (period III); and 5 min following the administration of 4,000 units of unfractionated heparin using a local delivery catheter system (period IV). We found consistent increases in both systemic arterial (P = 0.006) and coronary sinus (P = 0.0002) ATIII activity with systemic heparinization designed to prolong the activated clotting time to 300 sec. However, local delivery of heparin further increased coronary sinus ATIII activity (P = 0.003, period III vs. period IV). FpA concentration decreased in both systemic arterial (P < 0.0001) and coronary sinus (P < 0. 0001) samples following systemic heparinization. Moreover, local delivery of heparin further decreased coronary sinus FpA concentration (P = 0.04). Thus, on a background of intense anticoagulation during PTCA, the local delivery of 4,000 units of unfractionated heparin confers incremental antithrombotic activity. Cathet. Cardiovasc. Intervent. 48:84-88, 1999. PMID- 10467079 TI - Successful treatment of a bifurcation lesion with the Carina Bard stent: A case report. AB - In the last years new techniques and user-friendly stents have improved the results of stent implantation in coronary bifurcational lesions but difficulties in stent deployment and incomplete coverage of the bifurcation remain a problem. We describe the case of a proximal lesion of the anterior descending artery involving a large diagonal branch treated with the Carina Bard bifurcate stent, a new device that allows the complete immediate coverage of a bifurcational lesion with a single stent. Cathet. Cardiovasc. Intervent. 48:89-92, 1999. PMID- 10467080 TI - Push-pull angioplasty: ACE balloon-facilitated stent passage technique. AB - Improvements in interventional cardiology equipment have facilitated the treatment of more complex anatomies. Bulky stent transport systems and the inflexibility of stents have remained a limitation to the delivery of stents to distal, angulated, stiff, and calcified vessels. In order to provide the strongest guide backup, some vessel straightening, and a stiff rail over which the stent apparatus could be passed, an ACE balloon was advanced past the target lesion and used to trap the distal wire by gently inflating the ACE balloon at low pressure. Traction is then placed on the ACE catheter and the angioplasty guide wire at the touey valve insertion site. This procedure facilitates easy passage of angioplasty devices through challenging anatomies. In addition to the two case reports described, this technique has been used numerous other times with consistent success. Cathet. Cardiovasc. Intervent. 48:93-95, 1999. PMID- 10467081 TI - Sealing of coronary artery aneurysm by using a new stent graft. AB - Coronary stenting has begun to play an increasingly important role in the management of coronary artery aneurysms. A case of successful and complete sealing of a coronary aneurysm by using a new stent graft is described. Further studies in a large patient population are required to confirm the safety and efficacy of this method. Cathet. Cardiovasc. Intervent. 48:96-99, 1999. PMID- 10467082 TI - Use of the Shinobi wire in chronic coronary occlusions. AB - Occlusion angioplasty has represented a challenge since the introduction of the procedure. We report the successful use of a new, stiff, highly torquable, Teflon coated wire, the Shinobi wire, in nine coronary occlusions and have included the detailed description of three representative cases. In the discussion we have reviewed the current literature on devices used in coronary occlusions. Cathet. Cardiovasc. Intervent. 48:100-104, 1999. PMID- 10467083 TI - Stenting of long coronary artery lesions: initial angiographic results and 6 month clinical outcome of the micro stent II-XL. AB - To evaluate the results of long Micro Stent II (MS-XL) implantations, 119 MS-XLs were implanted in 102 patients (age, 62.83 years). Nineteen stents (16%) were implanted in saphenous vein grafts; 100 stents (84%) were implanted in native coronary arteries. Twenty-five patients (25%) were treated because of acute myocardial infarction (AMI); 30 patients (29%) because of unstable angina or angina class IV, and 47 patients (46%) because of stable angina. Eighty-six de novo lesions (84%) and 16 restenotic lesions (16%) were treated. Indications for stent implantation include elective, 61 patients (60%); suboptimal balloon angioplasty result, 22 patients (21%); and bailout after balloon angioplasty, 19 patients (19%). Because of residual thrombus after stenting, 27 patients (26%) received abciximab. All patients received ticlopidin for 28 days and acetylsalicylic acid. One hundred and seventeen MS-XLs (98%) were implanted successfully. Additional (shorter) MS-II were implanted in 40 patients (39%). The stented segment length was 45 +/- 20 mm. The minimum lumen diameter increased from 0.5 +/- 0.5 mm before to 2.7 +/- 0.5 mm after stent implantation. The acute gain was 2.2 +/- 0.4 mm. Early clinical events (<4 weeks) include death, 3 (3%); subacute stent thrombosis, 1 (1%); non-Q-wave infarction, 2 (2%); CABG, 1 (1%); vascular complications, 2 (2%). Late clinical events (<6 months) include acute myocardial infarction, 5 (5%); reintervention, 6 (6%); CABG, 1 (1%). The procedural success rate was 88%, and the event free survival at 6 months was 76%. Stenting of long lesions with the MS-XL was successful and associated with an acceptable complication rate. Cathet. Cardiovasc. Intervent. 48:105-112, 1999. PMID- 10467084 TI - Innovative interventions. PMID- 10467085 TI - Covered stents: home-made or ready-to-serve? PMID- 10467086 TI - Synchrotrons in the news. PMID- 10467087 TI - Holding damaged DNA together. AB - The mammalian X-ray cross-complementing group 1 protein (XRCC1) is an important player in base excision repair of damaged DNA. Two new findings help to elucidate its role - biochemical data suggest that this multidomain protein interacts not only with three different enzymes, but also with the nicked DNA itself, and NMR data reveal the structure of the domain that interacts with both DNA polymerase beta and DNA. PMID- 10467088 TI - More pieces of the K+ ion channel puzzle. AB - The secrets of biology's fastest switches - ion channels - are revealed. PMID- 10467089 TI - Growing momentum in the molecular study of ion channels. AB - Long the sole domain of physiologists and electrical engineers, ion channel biophysics is going molecular, and with a vengeance. This summary of a recent meeting devoted to ion channel biophysics shows that the integration of techniques, reinforced by active communication among scientists of diverse backgrounds, is extremely potent. PMID- 10467090 TI - Protein folding from 1961 to 1982. PMID- 10467091 TI - Picture story. Measuring motions in muscles. PMID- 10467092 TI - A novel target recognition revealed by calmodulin in complex with Ca2+-calmodulin dependent kinase kinase. AB - The structure of calcium-bound calmodulin (Ca2+/CaM) complexed with a 26-residue peptide, corresponding to the CaM-binding domain of rat Ca2+/CaM-dependent protein kinase kinase (CaMKK), has been determined by NMR spectroscopy. In this complex, the CaMKK peptide forms a fold comprising an alpha-helix and a hairpin like loop whose C-terminus folds back on itself. The binding orientation of this CaMKK peptide by the two CaM domains is opposite to that observed in all other CaM-target complexes determined so far. The N- and C-terminal hydrophobic pockets of Ca2+/CaM anchor Trp 444 and Phe 459 of the CaMKK peptide, respectively. This 14-residue separation between two key hydrophobic groups is also unique among previously determined CaM complexes. The present structure represents a new and distinct class of Ca2+/CaM target recognition that may be shared by other Ca2+/CaM-stimulated proteins. PMID- 10467093 TI - Confirmation of the hierarchical folding of RNase H: a protein engineering study. AB - The kinetic intermediate of RNase H is structured in a core region of the protein. To probe the role of this intermediate in the folding of RNase H, the folding kinetics of mutant proteins with altered native state stabilities were investigated. Mutations within the folding core destabilize the kinetic intermediate and slow refolding in a manner consistent with an obligatory intermediate model. Mutations outside of the folding core, however, do not affect the stability of the kinetic intermediate but do perturb the native state and transition state. These results indicate that interactions formed in the intermediate persist in the transition and native states and that RNase H folds through a hierarchical mechanism. PMID- 10467094 TI - Does trifluoroethanol affect folding pathways and can it be used as a probe of structure in transition states? AB - Nonaqueous co-solvents, particularly 2,2,2-trifluoroethanol (TFE), have been used as tools to study protein folding. By analyzing FKBP12, an alpha/beta-protein that folds with two-state kinetics, we have been able to address three key questions concerning the use of TFE. First, does TFE perturb the folding pathway? Second, can the observed changes in the rate of folding and unfolding in TFE be attributed to a change in free energy of a single state? Finally, can TFE be used to infer information on secondary structure formation in the transition state? Protein engineering experiments on FKBP12, coupled with folding and unfolding experiments in 0% and 9.6% TFE, conclusively show that TFE does not perturb the folding pathway of this protein. Our results also suggest that the changes in folding and unfolding rates observed in 9.6% TFE are due to a global effect of TFE on the protein, rather than the stabilization of any elements of secondary structure in the transition state. Thus, studies with TFE and other co-solvents can be accurately interpreted only when combined with other techniques. PMID- 10467095 TI - Structural basis for dimerization of the Dictyostelium gelation factor (ABP120) rod. AB - Gelation factor (ABP120) is one of the principal actin-cross-linking proteins of Dictyostelium discoideum. The extended molecule has an N-terminal 250-residue actin-binding domain and a rod constructed from six 100-residue repeats that have an Ig fold. The ability to dimerize is crucial to the actin cross-linking function of gelation factor and is mediated by the rod in which the two chains are arranged in an antiparallel fashion. We report the 2.2 A resolution crystal structure of rod domains 5 and 6, which shows that dimerization is mediated primarily by rod domain 6 and is the result of a double edge-to-edge extension of beta-sheets. Thus, contrary to earlier proposals, the chains of the dimeric gelation factor molecule overlap only within domain 6, and domains 1-5 do not pair with domains from the other chain. This information allows construction of a model of the gelation factor molecule and suggests how the chains in the related molecule filamin (ABP280) may interact. PMID- 10467096 TI - Structure of the 13-fold symmetric portal protein of bacteriophage SPP1. AB - We have determined the three-dimensional structure of bacteriophage SPP1 portal protein (gp6) using electron microscopy at liquid-helium temperatures and angular reconstitution. The 13-fold symmetric gp6 oligomer is a turbine-shaped structure with three distinct regions: a conical stem with a central channel; the turbine wings region; and a fringe of small 'tentacles' at the end of the channel exposed to the viral head interior. The tentacle region appears flexible and may be associated with a particular function - sensing when the correct amount of DNA has been packaged. The three-dimensional structure of the gp6 SizA mutant, which packages a smaller chromosome, reveals significant differences in that region. PMID- 10467097 TI - A productive NADP+ binding mode of ferredoxin-NADP + reductase revealed by protein engineering and crystallographic studies. AB - The flavoenzyme ferredoxin-NADP+ reductase (FNR) catalyzes the production of NADPH during photosynthesis. Whereas the structures of FNRs from spinach leaf and a cyanobacterium as well as many of their homologs have been solved, none of these studies has yielded a productive geometry of the flavin-nicotinamide interaction. Here, we show that this failure occurs because nicotinamide binding to wild type FNR involves the energetically unfavorable displacement of the C terminal Tyr side chain. We used mutants of this residue (Tyr 308) of pea FNR to obtain the structures of productive NADP+ and NADPH complexes. These structures reveal a unique NADP+ binding mode in which the nicotinamide ring is not parallel to the flavin isoalloxazine ring, but lies against it at an angle of approximately 30 degrees, with the C4 atom 3 A from the flavin N5 atom. PMID- 10467098 TI - Structure and mechanism of formation of the H-y5 isomer of an intramolecular DNA triple helix. AB - H-DNA, thought to play a regulatory role in transcription, exists in two isomeric forms, H-y3 and H-y5. We present the first solution structure of a DNA fragment representing the H-y5 fold. The structure shows the H-y5 triple helix, and for the first time how in an H-DNA isomer the purine strand extension interacts with the triplex loop. It gives direct insight into the mechanism of H-DNA formation, and explains a host of biochemical and biophysical data on the relative stability of the H-DNA isomers. In addition, the observed interaction of the purine strand extension and the triplex loop provides new clues to the design of clamp-type triple helix-forming oligonucleotides. PMID- 10467099 TI - Crystal structure of human heme oxygenase-1. AB - Heme oxygenase catalyzes the first step in the oxidative degradation of heme. The crystal structure of heme oxygenase-1 (HO-1) reported here reveals a novel helical fold with the heme sandwiched between two helices. The proximal helix provides a heme iron ligand, His 25. Conserved glycines in the distal helix near the oxygen binding site allow close contact between the helix backbone and heme in addition to providing flexibility for substrate binding and product release. Regioselective oxygenation of the alpha-meso heme carbon is due primarily to steric influence of the distal helix. PMID- 10467100 TI - Autoprocessing of HIV-1 protease is tightly coupled to protein folding. AB - In the Gag-Pol polyprotein of HIV-1, the 99-amino acid protease is flanked at its N-terminus by a transframe region (TFR) composed of the transframe octapeptide (TFP) and 48 amino acids of the p6pol, separated by a protease cleavage site. The intact precursor (TFP-p6pol-PR) has very low dimer stability relative to that of the mature enzyme and exhibits negligible levels of stable tertiary structure. Thus, the TFR functions by destabilizing the native structure, unlike proregions found in zymogen forms of monomeric aspartic proteases. Cleavage at the p6pol-PR site to release a free N-terminus of protease is concomitant with the appearance of enzymatic activity and formation of a stable tertiary structure that is characteristic of the mature protease as demonstrated by nuclear magnetic resonance. The release of the mature protease from the precursor can either occur in two steps at pH values of 4 to 6 or in a single step above pH 6. The mature protease forms a dimer through a four-stranded beta-sheet at the interface. Residues 1-4 of the mature protease from each subunit constitute the outer strands of the beta-sheet, and are essential for maintaining the stability of the free protease but are not a prerequisite for the formation of tertiary structure and catalytic activity. Our experimental results provide the basis for the model proposed here for the regulation of the HIV-1 protease in the viral replication cycle. PMID- 10467101 TI - A model of the changes in denatured state structure underlying m value effects in staphylococcal nuclease. AB - Hydrogen exchange kinetics were measured on the native states of wild type staphylococcal nuclease and four mutants with values of mGuHCl (defined as dDeltaG/d[guanidine hydrochloride]) ranging from 0.8 to 1.4 of the wild type value. Residues within the five-strand beta-barrel of wild type and E75A and D77A, two mutants with reduced values of m GuHCl, were significantly more protected from exchange than expected on the basis of global stability as measured by fluorescence. In contrast, mutants V23A and M26G with elevated values of mGuHCl approach a flat profile of more or less constant protection independent of position in the structure. Differences in exchange protection between the C terminus and the beta-barrel region correlate with mGuHCl, suggesting that a residual barrel-like structure becomes more highly populated in the denatured states of m- mutants and less populated in m+ mutants. Variations in the population of such a molten globule-like structure would account for the large changes in solvent accessible surface area of the denatured state thought to underlie m value effects. PMID- 10467102 TI - Solution structure of the single-strand break repair protein XRCC1 N-terminal domain. AB - XRCC1 functions in the repair of single-strand DNA breaks in mammalian cells and forms a repair complex with beta-Pol, ligase III and PARP. Here we describe the NMR solution structure of the XRCC1 N-terminal domain (XRCC1 NTD). The structural core is a beta-sandwich with beta-strands connected by loops, three helices and two short two-stranded beta-sheets at each connection side. We show, for the first time, that the XRCC1 NTD specifically binds single-strand break DNA (gapped and nicked). We also show that the XRCC1 NTD binds a gapped DNA-beta-Pol complex. The DNA binding and beta-Pol binding surfaces were mapped by NMR and found to be well suited for interaction with single-strand gap DNA containing a 90 degrees bend, and for simultaneously making contacts with the palm-thumb of beta-Pol in a ternary complex. The findings suggest a mechanism for preferential binding of the XRCC1 NTD to flexible single-strand break DNA. PMID- 10467103 TI - Heat shock activates the I-kappaBalpha promoter and increases I-kappaBalpha mRNA expression. AB - Recent data indicate that the heat shock response inhibits nuclear translocation of the proinflammatory transcription factor NF-kappaB. Under basal conditions NF kappaB is retained in the cytoplasm by an inhibitory protein called I-kappaB which exists as two major isoforms: I-kappaBalpha and I-kappaBbeta. Induction of the heat shock response in BEAS-2B cells, a human cell line representative of bronchial epithelium, increased expression of I-kappaBalpha mRNA in a time dependent manner. Coincubation with actinomycin-D inhibited heat shock-mediated expression of I-kappaBalpha mRNA. Transient transfection assays with a plasmid containing the reporter gene firefly luciferase, under the control of the human I kappaBalpha promoter, demonstrated that heat shock activated the I-kappaBalpha promoter. Heat shock-mediated induction of I-kappaBalpha was associated with inhibition of NF-kappaB activation. We conclude that heat shock increases I kappaBalpha mRNA expression in BEAS-2B cells by activating the I-kappaBalpha promoter, and propose that heat shock-mediated up-regulation of I-kappaBalpha is a potential mechanism by which the heat shock response inhibits proinflammatory responses in lung cells. PMID- 10467104 TI - Modulation of tolerance by mutant heat shock transcription factors. AB - It ought to be possible to recruit normal cellular defenses to mitigate ischemia/reperfusion damage and to reduce toxicity of chemotherapeutic drugs. Stress-preconditioned cells acquire a tolerant state characterized by increased resistance to such insults. This state is widely believed to be mediated, partially, by heat shock proteins (Hsps). Indirect evidence suggests that stress induced Hsp expression is controlled by heat shock transcription factor 1 (Hsf1), which factor may therefore represent a preferred target for therapeutic modulation of tolerance. In support, positively acting (Hsf1(+)) and negatively acting (Hsf1(-)) mutants of Hsf1 were identified. Inhibition of endogenous Hsf1 activity by Hsf1(-) prevents stress-induced Hsp synthesis and development of tolerance. Hsf1(+) drastically enhances expression of major Hsps in the absence of stress and induces tolerance against heat, simulated ischemia and toxicity by cyclophosphamide. Where compared, tolerance induced was slightly better than that produced by heat preconditioning. Thus, development of the tolerant state is dependent on increased levels of the cohort of Hsps induced by stress preconditioning, and Hsf1 can induce accumulation of a typical set of Hsps, which proteins are alone capable of providing tolerance at a similar level as heat preconditioning. These findings make Hsf1 a preferred target for pharmacological intervention to deliberately induce tolerance. PMID- 10467105 TI - Benzoquinoid ansamycins (herbimycin A and geldanamycin) interfere with the maturation of growth factor receptor tyrosine kinases. AB - Benzoquinoid ansamycins, such as herbimycin A (HA) and geldanamycin (GA), are antibiotics that exhibit anti-tumor effects. These compounds have been shown to result in the intracellular depletion of important growth signaling molecules. Recently, GA has been shown to bind tightly to Hsp90, thereby implicating Hsp90 as a possible chaperone for those signaling molecules adversely affected by the benzoquinoid ansamycins. Here we have investigated the effects of HA and GA on the synthesis, maturation and stability of different protein tyrosine kinases. Exposing cells to either compound blocked normal maturation of the epidermal growth factor (EGF) receptor, platelet-derived growth factor (PDGF) receptor, and pp60(v-src). We show that only the nascent forms of the EGF and PDGF receptors are degraded under these conditions. Once the newly synthesized receptors had been translocated into the endoplasmic reticulum membrane, addition of the drugs no longer affected their stability. For the cytoplasmic tyrosine kinase, pp60(v src), both the nascent as well as the mature forms of the protein were degraded in cells treated with the drugs. We discuss these observations as they pertain to the possible role of Hsp90 as a substrate-specific molecular chaperone, perhaps involved in the maturation and/or stability of proteins important for growth control. PMID- 10467106 TI - Identification of human heat shock protein 60 (Hsp60) and anti-Hsp60 antibodies in the peripheral circulation of normal individuals. AB - Although primarily regarded as being intracellular, this study has identified the presence of heat shock protein 60 (Hsp60) in the peripheral circulation of normal individuals. The median Hsp60 concentration was approximately 3.5-fold higher in females than in males and significantly higher levels of anti-human Hsp60 antibodies were also detected in females. There were no differences in the levels of antibodies to mycobacterial Hsp60 between males and females, nor did antibody levels correlate with Hsp60 concentrations. Hsp60 was not released from mitogenically stimulated peripheral blood mononuclear cells. The potential physiological roles for circulating Hsp60 are unknown. Given the emerging evidence that inappropriate reactivity to heat shock proteins is involved in autoimmune disease and that T cells responsive to self Hsp60 appear to be protective, these findings suggest that circulating Hsp60 may be involved in the regulation of tolerance to self and immunity to bacterial forms of these widely expressed and structurally conserved proteins. PMID- 10467107 TI - Activation of heat shock transcription factor 1 to a DNA binding form during the G(1)phase of the cell cycle. AB - The heat shock transcription factor (HSF) genes encode proteins that bind to the heat shock elements (HSE) of stress-inducible genes. We have observed the induction of HSF1, the ubiquitous member of the HSF family from a latent cytoplasmic state to a form competent to bind HSE during early G(1)in HeLa cells in the absence of stress. The induction of DNA-binding HSF1 coincided with a burst in cellular protein synthesis in early G(1)and inhibition of this translational peak prevented the formation of DNA binding-activated HSF1. A potential role for HSF1 in cell cycle regulation was suggested by the finding that cell lines stably overexpressing HSF1 showed an increased proportion of G(1)cells relative to other cell cycle phases. However, in contrast to the effects of heat shock, entry into G(1)did not lead to HSF1 hyperphosphorylation or increased activity of a heat shock promoter-reporter gene and did not cause the induction of heat shock protein 70 expression. Thus HSF1, previously implicated in the heat shock response is activated to a DNA binding from in G(1)under non-stress conditions and may play a role in G(1)regulation that does not involve the transcription of heat shock genes. PMID- 10467108 TI - Direct evidence for the intracellular localization of Hsp104 in Saccharomyces cerevisiae by immunoelectron microscopy. AB - To reveal the intracellular localization of Hsp104 in the yeast Saccharomyces cerevisiae before and after heat-shock, we performed immunoelectron microscopy after immunogold labeling with anti-Hsp104 antibody. At normal temperature (25 degrees C), a small amount of Hsp104 was located in the cytoplasm and nucleus. On exposure to mild heat-shock at 40 degrees C, protein aggregates appeared in the cytoplasm and nucleus, and Hsp104 increased around the aggregates with increasing time of the mild heat-shock treatment. Moreover, at lethal heat-shock temperature (51 degrees C) for 20 min after mild heat treatment at 40 degrees C, the intracellular localization of Hsp104 and intracellular structures were similar to those of the mild heat-shocked cells. However, in the lethally heat-shocked cells, certain intracellular structures were destroyed, and Hsp104 was not expressed. In the hsp104 null mutant strain Deltahsp104 which was treated at 40 degrees C, Hsp104 was not localized around the aggregates. Additionally, in the Deltahsp104 strain, even mild heat-shocked cells at 37 degrees C or 40 degrees C, showed destruction of intracellular structure compared to the wild-type strain. Our data suggest the following: (1) Hsp104 is associated closely with protein aggregates during heat-shock treatment, (2) Hsp104 is important for maintenance of the intracellular structure under lethal heat-shock conditions, (3) acquisition of thermotolerance depends on the amount of Hsp104 produced during mild heat-shock treatment. PMID- 10467109 TI - Meeting report on the International Congress on Hyperthermia in Clinical Oncology, Venice 1998. PMID- 10467110 TI - Highlights of the 1998 Cold Spring Harbor Meeting: Molecular chaperones and the heat shock response. PMID- 10467111 TI - Dynamics and regulation of the stress response: meeting report on the stress response and molecular chaperones from Kyoto. PMID- 10467113 TI - Ex vivo photodynamic purging in chronic myelogenous leukaemia and other neoplasias with rhodamine derivatives. AB - Photodynamic therapy (PDT), a cancer treatment already used early in this century, has distinctive advantages over conventional chemotherapy, namely its often observed preferential accumulation in cancer cells and its low intrinsic toxicity. Aggressive therapeutic modalities using high doses of chemotherapy and/or radiation therapy are now commonplace treatments for leukaemia, lymphoma and various non-haematologic malignancies. These intensive approaches have often been used in association with haematopoietic-progenitor-cell support and have induced major responses and remissions in patients with relapsed and refractory diseases, ultimately contributing to improve the disease-free survival of patients with high risk. This has encouraged Theratechnologies, a Montreal-based pharmaceutical company, to develop photodynamic ex vivo purging procedures, including the development of new photosensitizers and irradiation devices for the safe eradication of neoplastic cells from autologous grafts. Our first specific objective, therefore, was to design, synthesize, purify and test photoactive rhodamine derivatives. We have also selected a gas and phosphorus coating characteristic of an efficient scanning fluorescent source for extra-corporeal PDT using rhodamine derivatives. 4,5-Dibromorhodamine 123 (TH9402) was selected because of its photophysical properties, low toxicity and stability. TH9402 photodynamic-cell-therapy process conditions recognized as safe for normal human haematopoietic stem cells and progenitors demonstrated the efficacy of the purging procedure on various leukaemias (including chronic-myelogenous-leukaemia as well as non-Hodgkin-leukaemias and metastatic-breast-cancer cell lines. PMID- 10467114 TI - Purification and properties of Thermus filiformis DNA polymerase expressed in Escherichia coli. AB - The gene encoding Thermus filiformis (Tfi) DNA polymerase was expressed under the control of the tac promoter on a high-copy plasmid, pJR, in Escherichia coli. The Tfi DNA polymerase was purified by using heat treatment and DEAE-Sephacel column chromatography. The purified enzyme had a molecular mass of 92 kDa, as estimated by SDS/PAGE. The optimum pH and temperature of the enzyme were 8.4-9.0 and 70 72.5 degrees C respectively. The half-life of the enzyme at 94 degrees C was approx. 40 min. The enzyme was activated by the bivalent cations, Mg(2+) and Mn(2+), and was inhibited by EDTA. The optimal Mg(2+) concentration of the enzyme was 4 mM. The optimal conditions for the PCR reaction were slightly different from those for the enzyme activity except for the optimal Mg(2+) concentration. Low concentrations of KCl had no effect on either the enzymic activity or the PCR amplification. The result of the PCR experiment with the enzyme indicates that Tfi DNA polymerase might be useful in DNA amplification. PMID- 10467115 TI - Cholesterol oxidase from Streptomyces hygroscopicus and Brevibacterium sterolicum: effect of surfactants and organic solvents on activity. AB - We have studied systematically the effect of the non-ionic surfactants Thesit and Triton X-100, and of propan-2-ol (used as a substrate solubilizer) on the activity of the cholesterol oxidases from Streptomyces hygroscopicus (SCO) and Brevibacterium sterolicum (BCO). Low concentrations of Thesit lead to an activity increase with both enzymes; at higher surfactant concentrations the opposite effect occurs. Triton X-100 inactivates both enzymes at all concentrations. It is deduced that these surfactants exert their effects by interaction with the enzymes and not by affecting micellar phenomena. The effect of propan-2-ol on SCO, in contrast with that on BCO, depends on the buffer concentration (potassium phosphate). Other organic solvents induce results similar to those obtained with SCO and propan-2-ol. A significant difference between the two cholesterol oxidases emerges when stability is tested at 25 degrees C and in the presence of different concentrations of propan-2-ol: BCO activity is rapidly inactivated, whereas SCO still has 70% of the initial activity after 5 h in the presence of 30% propan-2-ol. From our results, SCO seems to be the catalyst of choice in comparison with BCO for the exploitation of cholesterol oxidases in biotechnology and applied biochemistry. PMID- 10467116 TI - Characterization of a novel stable biocatalyst obtained by protein engineering. AB - Protein engineering is a powerful tool for the improvement of the properties of biocatalysts. Previously we have applied protein engineering technologies to obtain an extremely stable variant of the thermolysin-like protease from Bacillus stearothermophilus [Van den Burg, Vriend, Veltman, Venema and Eijsink (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 2056-2060]. This variant is much more resistant to denaturing conditions (temperature and denaturing agents) than the wild-type enzyme. An extensive enzymic characterization was undertaken to explore the suitability of the variant as a biocatalyst at high temperatures. By comparing a range of variants with increasing thermal stabilities we show that the additivity of the mutations is accompanied by an increase in activity at elevated temperatures in accordance with the Arrhenius law. The results suggest that the constructed protease variants could be suitable alternatives to proteases that are currently used industrially. Our studies demonstrate how protein engineering can be exploited to obtain high-performance biocatalysts. PMID- 10467117 TI - Detection by HPLC of a trypanothione synthetase activity in vitro from Entamoeba histolytica. AB - We have previously demonstrated the presence of glutathione-spermidine (Gsp) and trypanothione [T(SH)(2)] from Entamoeba histolytica trophozoites, on the basis of results obtained with acid extracts purified by Florisil and DEAE-cellulose, derivatized with the fluorescent reagent monobromobimane and separated by HPLC. Gsp was originally found in Escherichia coli and later in trypanosomatids such as Trypanosoma cruzi, T. brucei, T. congolense and the insect trypanosomatid Crithidia fasciculata, along with the novel compound T(SH)(2), N(1), N(8) bis(glutathionyl)-spermidine. Here we demonstrate the presence of a T(SH)(2) synthetase activity in partly purified extracts from Entamoeba histolytica HK9, incubated at two pH values (6.5 and 7.5) with reduced glutathione (GSH), spermidine and ATP, in the presence of Mg(2+) at different time intervals. The thiol products were detected by HPLC in picomole amounts and compared with commercial Gsp and T(SH)(2) standards. We have used also an extract of Crithidia luciliae as a reference, to compare our results with C. fasciculata, in which the presence of this enzyme has previously been demonstrated and was later purified and separated into two synthetase activities from the same source: one for Gsp and the other for T(SH)(2). The presence of a T(SH)(2) synthetase activity in Entamoeba histolytica means that this protozoan has a similar metabolism to that of the trypanosomatids and opens the possibility of establishing a rational drug design against this human parasite. PMID- 10467118 TI - Entamoeba histolytica: a eukaryote with trypanothione metabolism instead of glutathione metabolism. AB - Entamoeba histolytica is a human pathogen that lacks the capacity to synthesize glutathione but can incorporate it, from the growth media or presumably from the human host, to form trypanothione [N(1), N(8)-bis(glutathionyl)-spermidine conjugate]. This novel thiol compound has previously been found in trypanosomatids, as has its precursor glutathionyl-spermidine, which was originally detected in Escherichia coli. Previously we showed the presence of these two thiol compounds in extracts from cultures of Entamoeba histolytica HK9. Here we report that when Entamoeba histolytica HK9 is grown in a culture medium that lacks glutathione (treated with the enzyme gamma-glutamyl transpeptidase), trypanothione is not formed, although the trophozoites can continue dividing for at least 60 h but at 25% lower cell density. The finding of a trypanothione metabolism in Entamoeba histolytica raises many questions: one concerns the possibility of a phylogenetic relationship, in this respect, with trypanosomatids such as Trypanosoma cruzi, T. brucei and Crithidia fasciculata; another concerns its role in cell metabolism; a third concerns it possible use as a target for a rational drug design strategy against this parasite. PMID- 10467119 TI - The exopolygalacturonase from Aspergillus tubingensis is also active on xylogalacturonan. AB - Apple-pectin hairy regions were prepared from apple pectin by combined action of the recombinant Aspergillus niger enzymes endopolygalacturonase II and pectin methylesterase and the A. tubigensis exopolygalacturonase. Using this enzymically prepared pectin fraction, an additional activity of the A. tubigensis exopolygalacturonase was discovered only when the substrate was chemically saponified and when D-galacturonate, a potent inhibitor of the enzyme, was removed from the incubation mixture. The new reaction product was purified and could be hydrolysed by A. niger beta-xylosidase into D-galacturonate and beta-D xylose in a 1:1 ratio, which identified it as xylogalacturonate. The results demonstrate that exopolygalacturonase is not only active on galacturonan but also on xylogalacturonan. The enzyme thus accomodates a substrate in which the terminal galacturonic acid residue carries a single xylose substitution. The well defined substrate specificity of exopolygalacturonase opens the possibility for use of this enzyme in biotechnological applications, such as preparing pectins that are methylated at the non-reducing end, and for studying the fine structure of xylogalacturonan in pectin. PMID- 10467120 TI - Expression and purification of a secreted functional mouse/human chimaeric antibody against bacterial endotoxin in baculovirus-infected insect cells. AB - We have created a mouse/human chimaeric antibody by taking antigen-binding fragment (Fab) genes of a mouse antibody-producing hybridoma with specificity for bacterial endotoxin and joining them to human Ig crystallizing-fragment (Fc) genes using recombinant DNA techniques. This chimaeric antibody has been expressed in Sf21 and High Fivetrade mark (BTI-TN-5B1-4) insect cells using the baculovirus expression system, which may allow the mass production of secretory recombinant antibodies. This was achieved by using infection with a double recombinant virus containing cDNAs of both the Ig heavy-chain (HC) and light chain (LC) genes. Prior to recombination, each gene was cloned into the dual expression baculovirus transfer vector pPLSP2, which permitted the insertion of the LC gene in-frame with the signal peptide of honey bee melittin downstream of the polyhedrin promoter, and of the HC gene in-frame with the signal peptide of Bombyx mori larval serum protein downstream of the p10 promoter. Our results showed that the polypeptide chains were secreted by insect cells and correctly assembled into H(2)L(2) heterodimers containing N-linked carbohydrate at the heavy chain. Furthermore, the recombinant chimaeric antibody exhibited a similar antigen specificity to that of the monoclonal antibody. More importantly, it provides a generic method for the high-level expression of antibodies. PMID- 10467121 TI - Anchorage-dependent and suspended baby-hamster kidney cells on three-dimensional non-woven polyester fabric discs: comparison of growth characteristics. AB - Two types of baby-hamster kidney (BHK) cell line, anchorage-dependent BHK-An(32) and suspended BHK-An(48) cells, were cultivated on non-woven polyester fabrics (NWPF) that were prepared from non-woven polyethylene teraphthalate sheets with different void fractions in three-dimensional configuration. The attachment and growth characteristics of both cell lines were investigated in stationary and submerged cultures, containing Dulbecco's modified Eagle's medium supplemented with fetal-calf serum. All cultures displayed rapid attachment (<120 min) of inoculated cells to the discs. The growth of BHK cells on NWPF discs in stationary cultures was similar to that observed in tissue-culture dishes; however, the denser discs showed better results in terms of final cell density, especially in the case of BHK-An(48) cells. Spinner-flask cultures indicated that higher cell yields were obtained than in stationary conditions. Microscopic observation showed that anchorage-dependent BHK-An(32) cells spread through the interfibre spaces of NPWF discs to form three-dimensional structures whereas BHK An(48) cells proliferated on fibres. PMID- 10467122 TI - Purification and biochemical characteristics of beta-D-xylanase from a thermophilic fungus, Thermomyces lanuginosus-SSBP. AB - An extracellular xylanase was purified to homogeneity from the culture filtrate of a thermophilic fungus, Thermomyces lanuginosus-SSBP, and its biochemical characteristics were studied. A yield of 70-80% was achieved through the procedures of 80%-satd. ammonium sulphate precipitation, DEAE-Sephadex A25 and quaternary aminoethyl (QAE)-Sephadex A25 column chromatography. The molecular mass of the purified xylanase was 23.6 kDa, as analysed by SDS/PAGE, with a pI value of 3.8. The molar absorption coefficient of the absorbance at 280 nm was 6.8x10(4) M(-1).cm(-1). The specific activity, calculated using the dinitrosalicylic acid (DNS) method, was 3500 units/mg. The enzyme reactions followed Michaelis-Menten kinetics with K app m and V(max) values of 3.26 mg/ml and 6300 units/ml per mg of protein respectively, as obtained from a Lineweaver Burk plot. The xylanase contained no other enzyme activity (cellulase, beta glucosidase, beta-mannosidase, alpha-arabinofuranosidase, or beta-xylosidase) except for the hydrolysis of xylan substrate. The optimal temperature of the enzyme assay was 70-75 degrees C. The enzyme retained full activity after a 60 degrees C incubation for 3 h. The optimal pH of xylanase activity was 6.5 and the enzyme appeared to be stable over a broad pH range (pH 5-12) under the assay conditions. The majority of the metal ions tested had no effect on the enzyme activity, with the exception of Pb(2+) (modest inhibitor) and Hg(2+) (strong inhibitor). The results showed that one or two tryptophan residues oxidized by N bromosuccinamide per enzyme molecule was sufficient to inhibit the enzyme activity completely, thus indicating that the tryptophan residues play an important role in the catalytical processes of the enzyme reaction. Because of the outstanding properties of the purified xylanase from the SSBP strain, this xylanase has a potential use in biopulping processes and other industrial applications. PMID- 10467123 TI - Purification and biochemical characteristics of beta-D-glucosidase from a thermophilic fungus, Thermomyces lanuginosus-SSBP. AB - The beta-D-glucosidase produced by Thermomyces lanuginosus-SSBP was purified to apparent homogeneity. The purified enzyme consisted of two identical subunits with a native molecular mass of 200 kDa. The purified beta-D-glucosidase only hydrolysed the glucoside substrates containing a terminal, non-reducing beta-D glucose residue and was active on both aryl-beta-glucoside and cellobiose. This enzyme also exhibited less, but significant alpha-D-glucosidase activity and was capable of hydrolysing beta-1,6-linked diglucosides and gentiobiose. The Kapp m, V(max) and k(cat) values for p-nitrophenyl-beta-D-glucopyranoside were calculated to be 0.075 mM, 12.12 units/mg of protein and 44.44 glucose molecules released/s respectively. The beta-D-glucosidase retained its full activity after a 30 min incubation at 50 degrees C but was inactive after the same treatment at 70 degrees C. The enzyme appeared to be stable when the pH of the storage buffer was above 5.0. Maximal beta-D-glucosidase activity occurred at 65 degrees C and pH 6.0. This enzyme was competitively inhibited by glucose, cellobiose and salicin with K(i) values of 0.55, 0.52 and 0.81 mM respectively. The presence of Hg(2+) and N-bromosuccinimide inhibited the enzyme activity completely at 2 mM, while cysteine enhanced beta-D-glucosidase activity. PMID- 10467124 TI - Signalling to actin: the Cdc42-N-WASP-Arp2/3 connection. AB - The molecular link between the signalling pathway regulating the formation of filopodia and the initiation of local actin polymerization has been elucidated: N WASP, a close homologue of WASP, which is the product of the gene responsible for the Wiskott-Aldrich syndrome, mediates a direct connection between the small G protein Cdc42 and the Arp2/3 complex. PMID- 10467125 TI - Ligand recognition by SH3 and WW domains: the role of N-alkylation in PPII helices. AB - SH3 and WW domains are involved in a variety of intracellular signaling pathways. Recent work has shed light on the mechanism whereby these signaling modules recognize prolines in polyproline ligands, which has implications in the design of ligands selectively targeting these interactions. PMID- 10467126 TI - RNA architecture dictates the conformations of a bound peptide. AB - BACKGROUND: The biological function of several viral and bacteriophage proteins, and their arginine-rich subdomains, involves RNA-mediated interactions. It has been shown recently that bound peptides adopt either beta-hairpin or alpha helical conformations in viral and phage peptide-RNA complexes. We have compared the structures of the arginine-rich peptide domain of HIV-1 Rev bound to two RNA aptamers to determine whether RNA architecture can dictate the conformations of a bound peptide. RESULTS: The core-binding segment of the HIV-1 Rev peptide class II RNA aptamer complex spans the two-base bulge and hairpin loop of the bound RNA and the carboxy-terminal segment of the bound peptide. The bound peptide is anchored in place by backbone and sidechain intermolecular hydrogen bonding and van der Waals stacking interactions. One of the bulge bases participates in U*(A*U) base triple formation, whereas the other is looped out and flaps over the bound peptide in the complex. The seven-residue hairpin loop is closed by a sheared G*A mismatch pair with several pyrimidines looped out of the hairpin fold. CONCLUSIONS: Our structural studies establish that RNA architecture dictates whether the same HIV-1 Rev peptide folds into an extended or alpha helical conformation on complex formation. Arginine-rich peptides can therefore adapt distinct secondary folds to complement the tertiary folds of their RNA targets. This contrasts with protein-RNA complexes in which elements of RNA secondary structure adapt to fit within the tertiary folds of their protein targets. PMID- 10467127 TI - Multiple regulatory genes in the tylosin biosynthetic cluster of Streptomyces fradiae. AB - BACKGROUND: The macrolide antibiotic tylosin is composed of a polyketide lactone substituted with three deoxyhexose sugars. In order to produce tylosin efficiently, Streptomyces fradiae presumably requires control mechanisms that balance the yields of the constituent metabolic pathways together with switches that allow for temporal regulation of antibiotic production. In addition to possible metabolic feedback and/or other signalling devices, such control probably involves interplay between specific regulatory proteins. Prior to the present work, however, no candidate regulatory gene(s) had been identified in S. fradiae. RESULTS: DNA sequencing has shown that the tylosin biosynthetic gene cluster, within which four open reading frames utilise the rare TTA codon, contains at least five candidate regulatory genes, one of which (tylP) encodes a gamma-butyrolactone signal receptor for which tylQ is a probable target. Two other genes (tylS and tylT) encode pathway-specific regulatory proteins of the Streptomyces antibiotic regulatory protein (SARP) family and a fifth, tylR, has been shown by mutational analysis to control various aspects of tylosin production. CONCLUSIONS: The tyl genes of S. fradiae include the richest collection of regulators yet encountered in a single antibiotic biosynthetic gene cluster. Control of tylosin biosynthesis is now amenable to detailed study, and manipulation of these various regulatory genes is likely to influence yields in tylosin-production fermentations. PMID- 10467128 TI - Heterologous expression, purification, reconstitution and kinetic analysis of an extended type II polyketide synthase. AB - BACKGROUND: Polyketide synthases (PKSs) are bacterial multienzyme systems that synthesize a broad range of natural products. The 'minimal' PKS consists of a ketosynthase, a chain length factor, an acyl carrier protein and a malonyl transferase. Auxiliary components (ketoreductases, aromatases and cyclases are involved in controlling the oxidation level and cyclization of the nascent polyketide chain. We describe the heterologous expression and reconstitution of several auxiliary PKS components including the actinorhodin ketoreductase (act KR), the griseusin aromatase/cyclase (gris ARO/CYC), and the tetracenomycin aromatase/cyclase (tcm ARO/CYC). RESULTS: The polyketide products of reconstituted act and tcm PKSs were identical to those identified in previous in vivo studies. Although stable protein-protein interactions were not detected between minimal and auxiliary PKS components, kinetic analysis revealed that the extended PKS comprised of the act minimal PKS, the act KR and the gris ARO/CYC had a higher turnover number than the act minimal PKS plus the act KR or the act minimal PKS alone. Adding the tcm ARO/CYC to the tcm minimal PKS also increased the overall rate. CONCLUSIONS: Until recently the principal strategy for functional analysis of PKS subunits was through heterologous expression of recombinant PKSs in Streptomyces. Our results corroborate the implicit assumption that the product isolated from whole-cell systems is the dominant product of the PKS. They also suggest that an intermediate is channeled between the various subunits, and pave the way for more detailed structural and mechanistic analysis of these multienzyme systems. PMID- 10467129 TI - Interactions of (-)-ilimaquinone with methylation enzymes: implications for vesicular-mediated secretion. AB - BACKGROUND: The marine sponge metabolite (-)-ilimaquinone has antimicrobial, anti HIV, anti-inflammatory and antimitotic activities, inhibits the cytotoxicity of ricin and diptheria toxin, and selectively fragments the Golgi apparatus. The range of activities demonstrated by this natural product provides a unique opportunity for studying these cellular processes. RESULTS: Affinity chromatography experiments show that (-)-ilimaquinone interacts with enzymes of the activated methyl cycle: S-adenosylmethionine synthetase, S adenosylhomocysteinase and methyl transferases. Known inhibitors of these enzymes were found to block vesicle-mediated secretion in a manner similar to (-) ilimaquinone. Moreover, the antisecretory effects of (-)-ilimaquinone and inhibitors of methylation chemistry, but not brefeldin A, could be reversed in the presence of the cellular methylating agent S-adenosylmethionine. Of the enzymes examined in the activated methyl cycle, S-adenosylhomocysteinase was specifically inhibited by (-)-ilimaquinone. Consistent with these observations, ( )-ilimaquinone was shown to obstruct new methylation events in adrenocorticotrophic hormone (ACTH)-secreting pituitary cells. CONCLUSIONS: (-) ilimaquinone inhibits cellular methylations through its interactions with S adenosylhomocysteinase. Furthermore, these studies indicate that the inhibition of secretion by ilimaquinone is the result of the natural product's antimethylation activity. It is likely that the ability to fragment the Golgi apparatus, as well as other activities, are also related to ilimaquinone's influence on methylation chemistry. PMID- 10467130 TI - Site-selective electron transfer from purines to electrocatalysts: voltammetric detection of a biologically relevant deletion in hybridized DNA duplexes. AB - BACKGROUND: The one-electron oxidation of guanine nucleobases is of interest for understanding the mechanisms of mutagenesis, probing electron-transfer reactions in DNA, and developing sensing schemes for nucleic acids. The electron-transfer rates for oxidation of guanine by exogenous redox catalysts depend on the base paired to the guanine. An important goal in developing the mismatch sensitivity is to identify a means for monitoring the current resulting from electron transfer at a single base in the presence of native oligonucleotides that contain all four bases. RESULTS: The nucleobase 8-oxo-guanine (8G) is selectively oxidized by the redox catalyst Os(bpy)(3)(3+/2+) (bpy = 2,2'-bipyridine) in the presence of native guanine. Cyclic voltammograms of Os(bpy)(3)(2+) show current enhancements indicative of nucleobase oxidation upon addition of oligonucleotides that contain 8G, but not in the presence of native guanine. As expected, similar experiments with Ru(bpy)(3)(2+) show enhancement with both guanine and 8G. The current enhancements for the 8G/Os(III) reaction increase in the order 8G-C approximately 8G.T < 8G.G < 8G.A < 8G, the same order as that observed for guanine/Ru(III). This site-selective mismatch sensitivity can be applied to detection of a TTT deletion, which is important in cystic fibrosis. CONCLUSIONS: The base 8G can be effectively used in conjunction with a low-potential redox catalyst as a probe for selective electron transfer at a single site. Because of the high selectivity for 8G, rate constants can be obtained that reflect the oxidation of only one base. The mismatch sensitivity can be used to detect biologically relevant abnormalities in DNA. PMID- 10467131 TI - Unusual stability of a multiply nicked form of Plasmodium falciparum triosephosphate isomerase. AB - BACKGROUND: The limited proteolytic cleavage of proteins can result in distinct polypeptides that remain noncovalently associated so that the structural and biochemical properties of the 'nicked' protein are virtually indistinguishable from those of the native protein. The remarkable observation that rabbit muscle triosephosphate isomerase (TIM) can be multiply nicked by subtilisin and efficiently religated in the presence of an organic solvent formed the stimulus for our study on a homologous system, Plasmodium falciparum triosephosphate isomerase (PfTIM). RESULTS: The subtilisin nicked form of PfTIM was prepared by limited proteolysis using subtilisin and the major fragments identified using electrospray ionization mass spectrometry. The order of susceptibility of the peptide bonds in the protein was also determined. The structure of the nicked form of TIM was investigated using circular dichroism, fluorescence and gel filtration. The nicked enzyme exhibited remarkable stability to denaturants, although significant differences were observed with the wild-type enzyme. Efficient religation could be achieved by addition of an organic cosolvent, such as acetonitrile, in the presence of subtilisin. Religation was also demonstrated after dissociation of the proteolytic fragments in guanidinium chloride, followed by reassembly after removal of the denaturant. CONCLUSIONS: The eight-stranded beta8/alpha8 barrel is a robust, widely used protein structural motif. This study demonstrates that the TIM barrel can withstand several nicks in the polypeptide backbone with a limited effect on its structure and stability. PMID- 10467132 TI - Secondary ligands enhance affinity at a designed metal-binding site. AB - BACKGROUND: Specific interactions of metal ions with proteins are central to all life processes. The varied functions enabled by this cooperation are a consequence of strict control of the binding-site environment, particularly the number, type and geometry of metal-coordinating sidechains. Attempts to mimic these characteristics in the de novo design of metal-binding sites have thus far concentrated primarily on metal recruitment and not on affecting site function through systematic fine-tuning of the metal environment. RESULTS: A designed tetrahedral Zn(II)-binding site in a variant of the B1 domain of IgG-binding protein G has been expanded by introducing 'secondary ligands'. These interactions were engineered to stabilize the positions of the metal-coordinating histidine residues while retaining the desired coordination geometry. Each mutation increased the protein's affinity for metal, and combining two secondary ligands demonstrated that these enhancements are additive. These results mimic the effects of altering similar interactions observed in the native Zn(II) binding site of carbonic anhydrase. In the B1 system, this enhanced affinity for metal is observed despite a substantial decrease in protein secondary structure. CONCLUSIONS: The intended effects of secondary ligand addition on metal affinity were observed in each mutant and demonstrated to be additive. Addition of metal also stabilized the protein's structure, partially offsetting the destabilizing effect of the mutations. These results represent a successful first attempt at designing an extended metal-binding site environment and illustrate the importance of including secondary interactions in the design of metal-binding sites. PMID- 10467134 TI - Re-introducing protein structure PMID- 10467133 TI - Structural basis for selective inhibition of Src family kinases by PP1. AB - BACKGROUND: Small-molecule inhibitors that can target individual kinases are powerful tools for use in signal transduction research. It is difficult to find such compounds because of the enormous number of protein kinases and the highly conserved nature of their catalytic domains. Recently, a novel, potent, Src family selective tyrosine kinase inhibitor was reported (PP1). Here, we study the structural basis for this inhibitor's specificity for Src family kinases. RESULTS: A single residue corresponding to Ile338 (v-Src numbering; Thr338 in c Src) in Src family tyrosine kinases largely controls PP1's ability to inhibit protein kinases. Mutation of Ile338 to a larger residue such as methionine or phenylalanine in v-Src makes this inhibitor less potent. Conversely, mutation of Ile338 to alanine or glycine increases PP1's potency. PP1 can inhibit Ser/Thr kinases if the residue corresponding to Ile338 in v-Src is mutated to glycine. We have accurately predicted several non-Src family kinases that are moderately (IC(50) approximately 1 microM) inhibited by PP1, including c-Abl and the MAP kinase p38. CONCLUSIONS: Our mutagenesis studies of the ATP-binding site in both tyrosine kinases and Ser/Thr kinases explain why PP1 is a specific inhibitor of Src family tyrosine kinases. Determination of the structural basis of inhibitor specificity will aid in the design of more potent and more selective protein kinase inhibitors. The ability to desensitize a particular kinase to PP1 inhibition of residue 338 or conversely to sensitize a kinase to PP1 inhibition by mutation should provide a useful basis for chemical genetic studies of kinase signal transduction. PMID- 10467135 TI - Peptide ligation and its application to protein engineering. AB - The ability to assemble a target protein from a series of peptide fragments, either synthetic or biosynthetic in origin, enables the covalent structure of a protein to be modified in an unprecedented fashion. The present technologies available for performing such peptide ligations are discussed, with an emphasis on how these methodologies have been utilized in protein engineering to investigate biological processes. PMID- 10467136 TI - Crystal structure of human mitochondrial NAD(P)(+)-dependent malic enzyme: a new class of oxidative decarboxylases. AB - Background: Malic enzymes catalyze the oxidative decarboxylation of malate to pyruvate and CO(2) with the concomitant reduction of NAD(P)(+) to NAD(P)H. They are widely distributed in nature and have important biological functions. Human mitochondrial NAD(P)(+)-dependent malic enzyme (mNAD-ME) may have a crucial role in the metabolism of glutamine for energy production in rapidly dividing cells and tumors. Moreover, this isoform is unique among malic enzymes in that it is a cooperative enzyme, and its activity is controlled allosterically. Results: The crystal structure of human mNAD-ME has been determined at 2.5 A resolution by the selenomethionyl multiwavelength anomalous diffraction method and refined to 2.1 A resolution. The structure of the monomer can be divided into four domains; the active site of the enzyme is located in a deep cleft at the interface between three of the domains. Three acidic residues (Glu255, Asp256 and Asp279) were identified as ligands for the divalent cation that is required for catalysis by malic enzymes. Conclusions: The structure reveals that malic enzymes belong to a new class of oxidative decarboxylases. The tetramer of the enzyme appears to be a dimer of dimers. The active site of each monomer is located far from the tetramer interface. The structure also shows the binding of a second NAD(+) molecule in a pocket 35 A away from the active site. The natural ligand for this second binding site may be ATP, an allosteric inhibitor of the enzyme. PMID- 10467137 TI - Identification of the Archaeoglobus fulgidus endonuclease III DNA interaction surface using heteronuclear NMR methods. AB - BACKGROUND: Endonuclease III is the prototype for a family of DNA-repair enzymes that recognize and remove damaged and mismatched bases from DNA via cleavage of the N-glycosidic bond. Crystal structures for endonuclease III, which removes damaged pyrimidines, and MutY, which removes mismatched adenines, show a highly conserved structure. Although there are several models for DNA binding by this family of enzymes, no experimental structures with bound DNA exist for any member of the family. RESULTS: Nuclear magnetic resonance (NMR) spectroscopy chemical shift perturbation of backbone nuclei (1H, 15N, 13CO) has been used to map the DNA-binding site on Archaeoglobus fulgidus endonuclease III. The experimentally determined interaction surface includes five structural elements: the helix hairpin-helix (HhH) motif, the iron-sulfur cluster loop (FCL) motif, the pseudo helix-hairpin-helix motif, the helix B-helix C loop, and helix H. The elements form a continuous surface that spans the active site of the enzyme. CONCLUSIONS: The enzyme-DNA interaction surface for endonuclease III contains five elements of the protein structure and suggests that DNA damage recognition may require several specific interactions between the enzyme and the DNA substrate. Because the target DNA used in this study contained a generic apurinic/apyrimidinic (AP) site, the binding interactions we observed for A. fulgidus endonuclease III should apply to all members of the endonuclease III family and several interactions could apply to the endonuclease III/AlkA (3-methyladenine DNA glycosylase) superfamily. PMID- 10467138 TI - Elucidating the medium-resolution structure of ribosomal particles: an interplay between electron cryo-microscopy and X-ray crystallograhy. AB - BACKGROUND: Ribosomes are the universal cellular organelles that accomplish the translation of the genetic code into proteins. Electron cryo-microscopy (cryo-EM) has yielded fairly detailed three-dimensional reconstructions of ribosomes. These were used to assist in the determination of higher resolution structures by X-ray crystallography. RESULTS: Molecular replacement studies using cryo-EM reconstructions provided feasible packing schemes for crystals of ribosomes and their two subunits from Thermus thermophilus, and of the large subunits from Haloarcula marismortui. For the large subunits, these studies also confirmed the major heavy-atom sites obtained by single isomorphous replacement combined with anomalous diffraction (SIRAS) and by multiple isomorphous replacement combined with anomalous diffraction (MIRAS) at approximately 10 A. Although adequate starting phases could not be obtained for the small subunits, the crystals of which diffract to 3.0 A, cryo-EM reconstructions were indispensable for analyzing their 7.2 A multiple isomorphous replacement (MIR) map. This work indicated that the conformation of the crystallized small subunits resembles that seen within the 70S ribosomes. Subsequently, crystals of particles trapped in their functionally active state were grown. CONCLUSIONS: Single-particle cryo-EM can contribute to the progress of crystallography of non-symmetrical, large and flexible macromolecular assemblies. Besides confirming heavy-atom sites, obtained from flat or overcrowded difference Patterson maps, the cryo-EM reconstructions assisted in elucidating packing arrangements. They also provided tools for the identification of the conformation within the crystals and for the estimation of the level of inherent non-isomorphism. PMID- 10467139 TI - A model for the incorporation of metal from the copper chaperone CCS into Cu,Zn superoxide dismutase. AB - BACKGROUND: Recent studies have identified the human copper chaperone CCS as the presumed factor responsible for copper incorporation into superoxide dismutase (SOD). A lack of knowledge of the chaperone's three-dimensional structure has prevented understanding of how the copper might be transferred. RESULTS: The three-dimensional structure of CCS was homology modelled using the periplasmic protein from the bacterial mercury-detoxification system and the structure of one subunit of the human SOD dimeric enzyme as templates. On the basis of the three dimensional model, a mechanism for the transfer of copper from CCS to SOD is proposed that accounts for electrostatic acceptor recognition, copper storage and copper-transfer properties. CONCLUSIONS: The proposed model identifies a path for copper transfer based on the presence of different metal sites characterized by sulphur ligands. Such a model permits the development of strategies able to interfere with copper incorporation in SOD, providing a possible way to prevent or arrest degeneration in the fatal motor neuron disorder amyotrophic lateral sclerosis. PMID- 10467140 TI - A new mode of B12 binding and the direct participation of a potassium ion in enzyme catalysis: X-ray structure of diol dehydratase. AB - BACKGROUND: Diol dehydratase is an enzyme that catalyzes the adenosylcobalamin (coenzyme B12) dependent conversion of 1,2-diols to the corresponding aldehydes. The reaction initiated by homolytic cleavage of the cobalt-carbon bond of the coenzyme proceeds by a radical mechanism. The enzyme is an alpha2beta2gamma2 heterooligomer and has an absolute requirement for a potassium ion for catalytic activity. The crystal structure analysis of a diol dehydratase-cyanocobalamin complex was carried out in order to help understand the mechanism of action of this enzyme. RESULTS: The three-dimensional structure of diol dehydratase in complex with cyanocobalamin was determined at 2.2 A resolution. The enzyme exists as a dimer of heterotrimers (alphabetagamma)2. The cobalamin molecule is bound between the alpha and beta subunits in the 'base-on' mode, that is, 5,6 dimethylbenzimidazole of the nucleotide moiety coordinates to the cobalt atom in the lower axial position. The alpha subunit includes a (beta/alpha)8 barrel. The substrate, 1,2-propanediol, and an essential potassium ion are deeply buried inside the barrel. The two hydroxyl groups of the substrate coordinate directly to the potassium ion. CONCLUSIONS: This is the first crystallographic indication of the 'base-on' mode of cobalamin binding. An unusually long cobalt-base bond seems to favor homolytic cleavage of the cobalt-carbon bond and therefore to favor radical enzyme catalysis. Reactive radical intermediates can be protected from side reactions by spatial isolation inside the barrel. On the basis of unique direct interactions between the potassium ion and the two hydroxyl groups of the substrate, direct participation of a potassium ion in enzyme catalysis is strongly suggested. PMID- 10467141 TI - Influence of nucleosome structure on the three-dimensional folding of idealized minichromosomes. AB - BACKGROUND: The closed circular, multinucleosome-bound DNA comprising a minichromosome provides one of the best known examples of chromatin organization beyond the wrapping of the double helix around the core of histone proteins. This higher level of chain folding is governed by the topology of the constituent nucleosomes and the spatial disposition of the intervening protein-free DNA linkers. RESULTS: By simplifying the protein-DNA assembly to an alternating sequence of virtual bonds, the organization of a string of nucleosomes on the minichromosome can be treated by analogy to conventional chemical depictions of macromolecular folding in terms of the bond lengths, valence angles, and torsions of the chain. If the nucleosomes are evenly spaced and the linkers are sufficiently short, regular minichromosome structures can be identified from analytical expressions that relate the lengths and angles formed by the virtual bonds spanning the nucleosome-linker repeating units to the pitch and radius of the organized quaternary structures that they produce. CONCLUSIONS: The resulting models with 4-24 bound nucleosomes illustrate how a minichromosome can adopt the low-writhe folding motifs deduced from biochemical studies, and account for published images of the 30 nm chromatin fiber and the simian virus 40 (SV40) nucleohistone core. The marked sensitivity of global folding to the degree of protein-DNA interactions and the assumed nucleosomal shape suggest potential mechanisms for chromosome rearrangements upon histone modification. PMID- 10467142 TI - Crystal structure of Pseudomonas fluorescens 4-hydroxyphenylpyruvate dioxygenase: an enzyme involved in the tyrosine degradation pathway. AB - BACKGROUND: In plants and photosynthetic bacteria, the tyrosine degradation pathway is crucial because homogentisate, a tyrosine degradation product, is a precursor for the biosynthesis of photosynthetic pigments, such as quinones or tocophenols. Homogentisate biosynthesis includes a decarboxylation step, a dioxygenation and a rearrangement of the pyruvate sidechain. This complex reaction is carried out by a single enzyme, the 4-hydroxyphenylpyruvate dioxygenase (HPPD), a non-heme iron dependent enzyme that is active as a homotetramer in bacteria and as a homodimer in plants. Moreover, in humans, a HPPD deficiency is found to be related to tyrosinemia, a rare hereditary disorder of tyrosine catabolism. RESULTS: We report here the crystal structure of Pseudomonas fluorescens HPPD refined to 2.4 A resolution (Rfree 27.6%; R factor 21.9%). The general topology of the protein comprises two barrel-shaped domains and is similar to the structures of Pseudomonas 2,3-dihydroxybiphenyl dioxygenase (DHBD) and Pseudomonas putida catechol 2,3-dioxygenase (MPC). Each structural domain contains two repeated betaalpha betabeta betaalpha modules. There is one non-heme iron atom per monomer liganded to the sidechains of His161, His240, Glu322 and one acetate molecule. CONCLUSIONS: The analysis of the HPPD structure and its superposition with the structures of DHBD and MPC highlight some important differences in the active sites of these enzymes. These comparisons also suggest that the pyruvate part of the HPPD substrate (4 hydroxyphenylpyruvate) and the O2 molecule would occupy the three free coordination sites of the catalytic iron atom. This substrate-enzyme model will aid the design of new inhibitors of the homogentisate biosynthesis reaction. PMID- 10467143 TI - Protein, lipid and water organization in bacteriorhodopsin crystals: a molecular view of the purple membrane at 1.9 A resolution. AB - BACKGROUND: Bacteriorhodopsin (bR) from Halobacterium salinarum is a proton pump that converts the energy of light into a proton gradient that drives ATP synthesis. The protein comprises seven transmembrane helices and in vivo is organized into purple patches, in which bR and lipids form a crystalline two dimensional array. Upon absorption of a photon, retinal, which is covalently bound to Lys216 via a Schiff base, is isomerized to a 13-cis,15-anti configuration. This initiates a sequence of events - the photocycle - during which a proton is transferred from the Schiff base to Asp85, followed by proton release into the extracellular medium and reprotonation from the cytoplasmic side. RESULTS: The structure of bR in the ground state was solved to 1.9 A resolution from non-twinned crystals grown in a lipidic cubic phase. The structure reveals eight well-ordered water molecules in the extracellular half of the putative proton translocation pathway. The water molecules form a continuous hydrogen-bond network from the Schiff-base nitrogen (Lys216) to Glu194 and Glu204 and includes residues Asp85, Asp212 and Arg82. This network is involved both in proton translocation occurring during the photocycle, as well as in stabilizing the structure of the ground state. Nine lipid phytanyl moieties could be modeled into the electron-density maps. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis of single crystals demonstrated the presence of four different charged lipid species. CONCLUSIONS: The structure of protein, lipid and water molecules in the crystals represents the functional entity of bR in the purple membrane of the bacteria at atomic resolution. Proton translocation from the Schiff base to the extracellular medium is mediated by a hydrogen-bond network that involves charged residues and water molecules. PMID- 10467144 TI - The 'mystery' of growing crystals of biomacromolecules comes to light. PMID- 10467145 TI - Protein folding in a specialized compartment: the endoplasmic reticulum. AB - The endoplasmic reticulum ensures proper folding of secretory proteins. In this review, we summarize and discuss the functions of different classes of folding mediators in the secretory pathway and propose updated models of the quality control system. PMID- 10467146 TI - Glutamate mutase from Clostridium cochlearium: the structure of a coenzyme B12 dependent enzyme provides new mechanistic insights. AB - BACKGROUND: Glutamate mutase (Glm) equilibrates (S)-glutamate with (2S,3S)-3 methylaspartate. Catalysis proceeds with the homolytic cleavage of the organometallic bond of the cofactor to yield a 5'-desoxyadenosyl radical. This radical then abstracts a hydrogen atom from the protein-bound substrate to initiate the rearrangement reaction. Glm from Clostridium cochlearium is a heterotetrameric molecule consisting of two sigma and two epsilon polypeptide chains. RESULTS: We have determined the crystal structures of inactive recombinant Glm reconstituted with either cyanocobalamin or methylcobalamin. The molecule shows close similarity to the structure of methylmalonyl CoA mutase (MCM), despite poor sequence similarity between its catalytic epsilon subunit and the corresponding TIM-barrel domain of MCM. Each of the two independent B12 cofactor molecules is associated with a substrate-binding site, which was found to be occupied by a (2S,3S)-tartrate ion. A 1:1 mixture of cofactors with cobalt in oxidation states II and III was observed in both crystal structures of inactive Glm. CONCLUSIONS: The long axial cobalt-nitrogen bond first observed in the structure of MCM appears to result from a contribution of the species without upper ligand. The tight binding of the tartrate ion conforms to the requirements of tight control of the reactive intermediates and suggests how the enzyme might use the substrate-binding energy to initiate cleavage of the cobalt-carbon bond. The cofactor does not appear to have a participating role during the radical rearrangement reaction. PMID- 10467147 TI - An immunoglobulin-like fold in a major plant allergen: the solution structure of Phl p 2 from timothy grass pollen. AB - BACKGROUND: Grass pollen allergens are the most important and widespread elicitors of pollen allergy. One of the major plant allergens which millions of people worldwide are sensitized to is Phl p 2, a small protein from timothy grass pollen. Phl p 2 is representative of the large family of cross-reacting plant allergens classified as group 2/3. Recombinant Phl p 2 has been demonstrated by immunological cross-reactivity studies to be immunologically equivalent to the natural protein. RESULTS: We have solved the solution structure of recombinant Phl p 2 by means of nuclear magnetic resonance techniques. The three-dimensional structure of Phl p 2 consists of an all-beta fold with nine antiparallel beta strands that form a beta sandwich. The topology is that of an immunoglobulin-like fold with the addition of a C-terminal strand, as found in the C2 domain superfamily. Lack of functional and sequence similarity with these two families, however, suggests an independent evolution of Phl p 2 and other homologous plant allergens. CONCLUSIONS: Because of the high homology with other plant allergens of groups 1 and 2/3, the structure of Phl p 2 can be used to rationalize some of the immunological properties of the whole family. On the basis of the structure, we suggest possible sites of interaction with IgE antibodies. Knowledge of the Phl p 2 structure may assist the rational structure-based design of synthetic vaccines against grass pollen allergy. PMID- 10467148 TI - Coagulation factor IXa: the relaxed conformation of Tyr99 blocks substrate binding. AB - BACKGROUND: Among the S1 family of serine proteinases, the blood coagulation factor IXa (fIXa) is uniquely inefficient against synthetic peptide substrates. Mutagenesis studies show that a loop of residues at the S2-S4 substrate-binding cleft (the 99-loop) contributes to the low efficiency. The crystal structure of porcine fIXa in complex with the inhibitor D-Phe-Pro-Arg-chloromethylketone (PPACK) was unable to directly clarify the role of the 99-loop, as the doubly covalent inhibitor induced an active conformation of fIXa. RESULTS: The crystal structure of a recombinant two-domain construct of human fIXa in complex with p aminobenzamidine shows that the Tyr99 sidechain adopts an atypical conformation in the absence of substrate interactions. In this conformation, the hydroxyl group occupies the volume corresponding to the mainchain of a canonically bound substrate P2 residue. To accommodate substrate binding, Tyr99 must adopt a higher energy conformation that creates the S2 pocket and restricts the S4 pocket, as in fIXa-PPACK. The energy cost may contribute significantly to the poor K(M) values of fIXa for chromogenic substrates. In homologs, such as factor Xa and tissue plasminogen activator, the different conformation of the 99-loop leaves Tyr99 in low-energy conformations in both bound and unbound states. CONCLUSIONS: Molecular recognition of substrates by fIXa seems to be determined by the action of the 99 loop on Tyr99. This is in contrast to other coagulation enzymes where, in general, the chemical nature of residue 99 determines molecular recognition in S2 and S3-S4. This dominant role on substrate interaction suggests that the 99-loop may be rearranged in the physiological fX activation complex of fIXa, fVIIIa, and fX. PMID- 10467149 TI - The start of a new generation: the present status of the SPring-8 synchrotron and its use in structural biology. PMID- 10467150 TI - Solution structure and dynamics of a designed hydrophobic core variant of ubiquitin. AB - BACKGROUND: The recent merger of computation and protein design has resulted in a burst of success in the generation of novel proteins with native-like properties. A critical component of this coupling between theory and experiment is a detailed analysis of the structures and stabilities of designed proteins to assess and improve the accuracy of design algorithms. RESULTS: Here we report the solution structure of a hydrophobic core variant of ubiquitin, referred to as 1D7, which was designed with the core-repacking algorithm ROC. As a measure of conformational specificity, we also present amide exchange protection factors and backbone and sidechain dynamics. The results indicate that 1D7 is similar to wild type (WT) ubiquitin in backbone structure and degree of conformational specificity. We also observe a good correlation between experimentally determined sidechain structures and those predicted by ROC. However, evaluation of the core sidechain conformations indicates that, in general, 1D7 has more sidechains in less statistically favorable conformations than WT. CONCLUSIONS: Our results provide an explanation for the lower stability of 1D7 compared to WT, and suggest modifications to design algorithms that may improve the accuracy with which structure and stability are predicted. The results also demonstrate that core packing can affect conformational flexibility in subtle ways that are likely to be important for the design of function and protein-ligand interactions. PMID- 10467151 TI - Crystal structure of an aromatic ring opening dioxygenase LigAB, a protocatechuate 4,5-dioxygenase, under aerobic conditions. AB - BACKGROUND: Sphingomonas paucimobilis SYK-6 utilizes an extradiol-type catecholic dioxygenase, the LigAB enzyme (a protocatechuate 4,5-dioxygenase), to oxidize protocatechuate (or 3,4-dihydroxybenzoic acid, PCA). The enzyme belongs to the family of class III extradiol-type catecholic dioxygenases catalyzing the ring opening reaction of protocatechuate and related compounds. The primary structure of LigAB suggests that the enzyme has no evolutionary relationship with the family of class II extradiol-type catecholic dioxygenases. Both the class II and class III enzymes utilize a non-heme ferrous center for adding dioxygen to the substrate. By elucidating the structure of LigAB, we aimed to provide a structural basis for discussing the function of class III enzymes. RESULTS: The crystal structure of substrate-free LigAB was solved at 2.2 A resolution. The molecule is an alpha2beta2 tetramer. The active site contains a non-heme iron coordinated by His12, His61, Glu242, and a water molecule located in a deep cleft of the beta subunit, which is covered by the alpha subunit. Because of the apparent oxidation of the Fe ion into the nonphysiological Fe(III) state, we could also solve the structure of LigAB complexed with a substrate, PCA. The iron coordination sphere in this complex is a distorted tetragonal bipyramid with one ligand missing, which is presumed to be the O2-binding site. CONCLUSIONS: The structure of LigAB is completely different from those of the class II extradiol type dioxygenases exemplified by the BphC enzyme, a 2,3-dihydroxybiphenyl 1,2 dioxygenase from a Pseudomonas species. Thus, as already implicated by the primary structures, no evolutionary relationship exists between the class II and III enzymes. However, the two classes of enzymes share many geometrical characteristics with respect to the nature of the iron coordination sphere and the position of a putative catalytic base, strongly suggesting a common catalytic mechanism. PMID- 10467152 TI - Pretravel advice neglects rabies risk for travelers to tropical countries. AB - BACKGROUND: While the risk of acquiring rabies in Europe is low, there have been a number of reports on cases of human rabies "imported" from abroad in recent years. Few studies have been conducted concerning pretravel advice for international travelers about the risk of rabies. METHODS: One-hundred and fifty German and 150 Swiss general practitioners (GPs) who give pretravel advice were interviewed to assess their awareness of the risk of rabies for travelers, and of the relevant preventive measures, using a pretested telephone interview followed by a written questionnaire with a multiple choice list. GPs were asked specifically about pretravel health advice for journeys to Thailand and Kenya. RESULTS: During the telephone interview a majority (76%) of the 300 GPs recommended "nothing" as a preventive measure against rabies for travelers. One or two important preventive measures were mentioned by only a few GPs. No major differences were detected between the German and the Swiss groups concerning their recommendations for rabies prevention. The 119 German and 121 Swiss GPs who returned the completed questionnaire said they would recommend the three important preventive measures in the future significantly more often than they had recommended them to their clients in the past. CONCLUSIONS: The risk of rabies in travelers to tropical countries appears to be neglected in pretravel advice provided by Swiss and German GPs. The recommendations on travel advice published in the Swiss Bulletin of the Federal Office of Public Health (BFOPH) and in the "Deutsches Arzteblatt" should be extended to "other specific health risks." In Switzerland this has now been done. PMID- 10467153 TI - Anatomy of a world cruise. AB - BACKGROUND AND METHOD: We registered the patient-related activities of a large cruise ship's medical center during a 103-day worldwide voyage and compared data from 694 passengers (53% women) with median age 66 years with data from 540 crew (27% women) with median age 30 years of age. RESULTS: The medical staff had 3033 (1537 crew, 1496 passenger) consultations (=206 consultations per week) and performed 982 diagnostic procedures. Skin disorders dominated in the crew and were more frequent than in the passengers (29% versus 13%, p<.01), respiratory problems dominated in the passengers and were more frequent than in the crew (26% versus 17%, p<.01), while cardiovascular disorders were more common in the passengers (7% versus 1%, p<.01). Forty-six injuries (35 passengers, 11 crew) were recorded. One passenger died aboard. Seventy-six crew (14%) were not able to work for a total of 110 days (1.1 crew sick per day), while five crew were signed off for medical reasons. CONCLUSIONS: To be a doctor or nurse on a large cruise ship during a long sea voyage is no vacation. The medical staff should expect a varied general practice with a higher consultation frequency rate than shoreside, even higher in crew PMID- 10467154 TI - Risk to tourists posed by wild mammals in South Africa. AB - BACKGROUND: One of South Africa's principal tourist attractions is the opportunity to encounter Africa's large mammals in the wild. Attacks by these mammals can be exceptionally newsworthy with potentially deleterious effects on tourism. Little is known about the risk of injury and death caused by wild mammals to visitors to South Africa's nature reserves. The aim of this study was to determine the incidence of fatal and nonfatal attacks on tourists by wild mammals in South Africa and to ascertain avoidable factors, if any. METHODS: Commercial press records covering all South African Newspapers archived at the Independent Newspapers' central library were systematically reviewed for a 10 year period, January 1988 to December 1997 inclusive, to identify all deaths and injuries to domestic and international tourists resulting from encounters with wild mammals in South Africa. All of these incidents were analyzed to ascertain avoidable factors. RESULTS: During the review period seven tourists, including two students from Thailand and a German traveler, were killed by wild mammals in South Africa. Three of the four deaths ascribed to lions resulted from tourists carelessly approaching prides on foot in lion reserves. A judicial inquiry found that the management of a KwaZulu-Natal Reserve was culpable for the remaining death. Tourist ignorance of animal behavior and flagrant disregard of rules contributed to the two fatalities involving hippopotami. The unusual behavior manifested by the bull elephant responsible for the final death, resulted from discomfort caused by a dental problem to this pachyderm. During the same period there were 14 nonfatal attacks on tourists, including five by hippo, three by buffalo, two by rhino, and one each by a lion, leopard, zebra and musth elephant. Only the latter occurred while the visitor was in a motor vehicle. Tourist ethological naivete and failure to determine the experience of trail guides prior to travel, resulted in inadvertent agonistic behavior, unnecessary risk-taking and avoidable injury. CONCLUSIONS: This retrospective study has shown that attacks on tourists by wild mammals in South Africa are an uncommon cause of injury and death. Sensible precautions to minimize this risk include remaining in a secure motor vehicle or adequately fenced precincts while in the vicinity of large mammals, rigidly observing nature reserve instructions, never approaching animals that appear ill, malnourished, displaying aggressive behavior traits or female wild mammals with young, and demanding adequately trained and experienced game rangers when embarking on walking trails. Any behavior that might be construed as antagonistic and which could provoke an attack by large mammals should be avoided (e.g., driving directly at a lion). Visitors need to be informed of classic signs of aggression, in particular in elephants, which will allow timely avoidance measures to be taken. The risk-enhancing effect of excessive alcohol intake is undesirable in the game reserve setting, as is driving at high speed after dusk in areas where hippos graze. Local advice on personal safety in wildlife reserves and the credentials of trail guides should be obtained from lodge or reserve management, tourism authorities or the travel industry prior to travel to game reserves. PMID- 10467155 TI - Diving and marine medicine review part II: diving diseases. AB - Diving is a high-risk sport. There are approximately between 1 to 3 million recreational scuba divers in the USA (with over a quarter-million learning scuba annually); there are about 1 million in Europe and over 50,000 in the United Kingdom. In this population 3-9 deaths/100,000 occur annually in the US alone, and those surviving diving injuries far exceeds this. Diving morbidity can be from near-drowning, from gas bubbles, from barotrauma or from environmental hazards. In reality, the most common cause of death in divers is drowning (60%), followed by pulmonary-related illnesses. The mean number of annual diving fatalities in the USA from 1970 to 1993 was 103.5 (sd 24.0) and the median was 106. This article will focus primarily upon pressure effects on the health of a diver. There are two principle ways pressure can affect us: by direct mechanical effects and by changing the partial pressures of inspired gases. Dysbarism is a general term used to describe pathology from altered environmental pressure, and has two main forms: barotrauma from the uncontrolled expansion of gas within gas filled body compartments and decompression sickness from too rapid a return to atmospheric pressure after breathing air under increased pressures. Greater than 90% of the human body is either water or bone, which is incompressible; the areas directly affected by pressure changes thus are those that are filled with air or gas. These sites include the middle ear, the eustachian tube, the sinuses, the thorax, and the gastrointestinal tract. Air in these cavities is compressed when the ambient pressure rises because the pressure of inhaled air must equilibrate with the ambient pressure. PMID- 10467156 TI - Health risks of travelers in South Africa. AB - Large numbers of tourists visit South Africa every year. Travelers to urban areas are at little risk of contracting an infectious disease, however the adventure traveler is at increased risk. Yellow fever is not known to occur in South Africa. Malaria is endemic in Mpumalanga and KwaZula-Natal. Schistosomiasis is endemic in large parts of the country. Although rabies is found throughout the country, only a small number of human cases is reported. High risk areas are KwaZulu-Natal, the eastern Cape and Mpumalanga provinces. The incidence of human immunodeficiency virus (HIV) infection is high and counseling regarding sexually transmitted diseases is important. Sanitation of water is excellent in most large cities and towns; however travelers to rural areas should exercise caution. Arbovirus infections do occur but relatively few cases are reported. The hiker is at risk for tick bite fever and should be counseled. Since the abolition of apartheid, South Africa has been seen as an inexpensive, high quality destination by many tourists. In 1997, a total of 5,436,848 travelers from many different countries visited the country. Areas most frequently visited include Johannesburg, Cape Town, Durban, the Garden Route, Kruger National Park, KwaZulu Natal and Pretoria. The most common reason for visiting the country was holiday (44%), followed by visiting friends and relatives (23%), business travel (27%) other (6%).1 Travelers, to the larger cities such as Johannesburg, Cape Town and Durban are at little risk of acquiring an infectious disease. The adventure traveler however is at greater risk as parts of the country are endemic for malaria, schistosomiasis, rabies, food and waterborne diseases, sexually transmitted diseases and arbovirus infections. Accidental deaths due to motor vehicle accidents and interpersonal violence are important health risks in South Africa. Travelers visiting popular attractions are at significantly lower risk. However this has never been quantified. This review aims to address the occurrence of infectious diseases and attempts to give guidelines to practitioners caring for travelers. PMID- 10467157 TI - Pediatric migration and hepatitis A risk in host population. AB - Hepatitis A virus (HAV) circulation in a given area is closely related to socioeconomic standards. Following the improvement of living conditions, HAV seroprevalence rates in the population have decreased steadily during the last decades in many Western European countries, including Italy, thereby leading to a shift of risk of disease towards older age groups. Since the severity of the disease closely parallels age, a higher incidence of symptomatic cases in adults is now reported in Europe and the United States, being travel-related to a large extent. Intrafamilial person-to-person spread is also an important source of infection and transmission from children to parents may occur due to the lack of immunity in the general population. In the last two decades, Italy has been the destination of an increasing number of migrants from developing countries, where HAV is highly endemic. Furthermore, international adoption programmes cause pediatric populations from HAV endemic countries to increase in low endemic areas, possibly leading to secondary cases in close contacts.7 The aim of this paper is to report the epidemic HAV outbreak which occurred among the voluntary nursing staff of a pediatric Rwandan refugee community hosted in a village of the Brescia Province, in northern Italy. PMID- 10467158 TI - An eighty-four-year-old man with fever and painless jaundice: a case report and brief review of Clonorchis sinensis infection. PMID- 10467159 TI - Effects of missense mutations Phe110Ile and Glu244Asp in human cardiac troponin T on force generation in skinned cardiac muscle fibers. AB - The functional effects of two missense mutations in human cardiac troponin T, Phe110Ile and Glu244Asp, associated with familial hypertrophic cardiomyopathy were examined by exchanging the bacterially expressed and purified mutant troponin T into rabbit cardiac skinned muscle fibers. Both mutations significantly increased the maximum force without affecting the cooperativity. The Glu244Asp mutation also increased the Ca(2+) sensitivity of the force generation, as in the case of other mutations associated with a poor prognosis. On the other hand, the Phe110Ile mutation, associated with a favorable prognosis, had no effect on the Ca(2+) sensitivity. The results strongly support the hypothesis that increased Ca(2+) sensitivity is responsible for the pathogenesis of hypertrophic cardiomyopathy with a poor prognosis caused by mutations in troponin T. PMID- 10467160 TI - Differential and additive effects of the three conserved isoleucine residues in the promoter -10 binding region on Bacillus subtilis sigma(A) structure and function. AB - The promoter -10 binding region of the Bacillus subtilis sigma(A) factor forms an amphiphilic alpha-helix with three conserved isoleucines located at four-residue intervals. To investigate the structural and functional roles of the three isoleucine residues, we constructed a series of sigA mutants with single and double Ile-to-Ala substitutions on the hydrophobic face of this alpha-helix and isolated intragenic revertants with either same-site or second-site suppressor that partially restores the structural stability and transcription activity of the mutant sigma(A) factors. Our data show that the three conserved isoleucine residues (Ile-194, Ile-198, and Ile-202) are involved in the hydrophobic core packing of sigma(A); they affect differentially and additively the structure and function of sigma(A), with the central isoleucine residue (Ile-198) playing the most important role. By analogy with the crystal structure of a sigma(70) peptide, it is apparent that interdigital interactions exist between the three conserved isoleucine residues and certain hydrophobic amino acids in region 2. 1 of sigma(A). They include at least the van der Waals contacts between Ile-194 and both Leu-145 and Ile-149, between Ile-198 and both Ile-149 and Tyr-153, as well as between Ile-202 and Tyr-153. The same-site suppressors, Val-194 and Val-198, restore the structural stability and transcription activity of sigma(A) by repacking the hydrophobic core of sigma(A). The second-site suppressor (S291F) appears to be allele-specific, but it is not as effective as the same-site suppressors in restoring sigma(A) structure and function. PMID- 10467161 TI - Chitosanase-catalyzed hydrolysis of 4-methylumbelliferyl beta-chitotrioside. AB - 4-Methylumbelliferyl beta-chitotrioside [(GlcN)(3)-UMB] was prepared from 4 methylumbelliferyl tri-N-acetyl-beta-chitotrioside [(GlcNAc)(3)-UMB] using chitin deacetylase from Colletotrichum lindemuthianum, and hydrolyzed by chitosanase from Streptomyces sp. N174. The enzymatic deacetylation of (GlcNAc)(3)-UMB was confirmed by (1)H-NMR spectroscopy and mass spectrometry. When the (GlcN)(3)-UMB obtained was incubated with chitosanase, the fluorescence intensity at 450 nm obtained by excitation at 360 nm was found to increase with proportion to the reaction time. The rate of increase in the fluorescence intensity was proportional to the enzyme concentration. This indicates that chitosanase hydrolyzes the glycosidic linkage between a GlcN residue and UMB moiety releasing the fluorescent UMB molecule. Since (GlcN)(3) itself cannot be hydrolyzed by the chitosanase, (GlcN)(3)-UMB is considered to be a useful low molecular weight substrate for the assay of chitosanase. The k(cat) and K(m) values obtained for the substrate (GlcN)(3)-UMB were determined to be 8.1 x 10(-5) s(-1) and 201 microM, respectively. From TLC analysis of the reaction products, the chitosanase was found to hydrolyze not only the linkages between a GlcN residue and UMB moiety, but also the linkages between GlcN residues. Nevertheless, the high sensitivity of the fluorescence detection of the UMB molecule would enable a more accurate determination of kinetic constants for chitosanases. PMID- 10467162 TI - Mutational analysis of the regulatory mechanism of PKN: the regulatory region of PKN contains an arachidonic acid-sensitive autoinhibitory domain. AB - PKN is a fatty acid- and Rho GTPase-activated protein kinase whose catalytic domain in the carboxyl terminus is homologous to those of protein kinase C (PKC) family members. The amino terminal region of PKN is suggested to function as a regulatory domain, since tryptic cleavage or the binding of Rho GTPase to this region results in protein kinase activation of PKN. The structural basis for the regulation of PKN was investigated by analyzing the activity of a series of deletion/site-directed mutants expressed in insect cells. The amino-terminally truncated form of PKN (residue 455-942) showed low basal activity similar to that of the wild-type enzyme, and was arachidonic acid-dependent. However, further deletion (residue 511-942) resulted in a marked increase in the basal activity and a decrease in the arachidonic acid dependency. A (His)(6)-tagged protein comprising residues 455-511 of PKN (designated His-Ialpha) inhibited the kinase activity of the catalytic fragment of PKN in a concentration-dependent manner in competition with substrate (K(i) = 0.6+/-0.2 microM). His-Ialpha also inhibited the activity of the catalytic fragment of PRK2, an isoform of PKN, but had no inhibitory effect on protein kinase A or protein kinase Cdelta. The IC(50) value obtained in the presence of 40 microM arachidonic acid was two orders of magnitude greater than that in the absence of the modifier. These results indicate that this protein fragment functions as a specific inhibitor of PKN and PRK2, and that arachidonic acid relieves the catalytic activity of wild-type PKN from autoinhibition by residues 455-511 of PKN. Autophosphorylation of wild-type PKN increased the protein kinase activity, however, substitution of Thr64, Ser374, or Thr531 in the regulatory region of PKN with alanine, abolished this effect. Substitution of Thr774 in the activation loop of the catalytic domain of PKN with alanine completely abolished the protein kinase activity. These results suggest that these phosphorylation sites are also important in the regulation of the PKN kinase activity. Potential differences in the mechanism of activation between the catalytic regions of PKN and PRK2 are also discussed. PMID- 10467163 TI - Human parathyroid hormone (1-34) transiently increases the excretion of lysosomal enzymes into urine and the size of renal lysosomes. AB - It has been reported that the urinary excretion of N-acetyl-beta-D glucosaminidase (NAG), a lysosomal enzyme, transiently increases in human after treatment with human parathyroid hormone (hPTH)(1-34). We report here that hPTH(1 34) caused transient changes in the size and density of rat renal lysosomes following urinary excretion of NAG and other lysosomal enzymes tested. Percoll density gradient centrifugation revealed that hPTH(1-34) slightly but significantly increased the fraction of high density lysosomes (around 1.12 g/ml) 5-10 min after the treatment with hPTH(1-34), with a concomitant decrease in the fraction of intermediate density lysosomes (1.07-1.08 g/ml). On electron micrographs, some lysosomes in proximal tubules but not in distal tubules showed a change in morphology from circular to oval, and became enlarged and electron dense 5-10 min after the treatment with hPTH(1-34). These responses to hPTH(1-34) were also reversible and transient. NAG excreted in urine after treatment with hPTH(1-34) had the molecular mass of a mature form in lysosomes and/or endosomes and was not a prepro-and/or pro-form of the enzyme. Thus, the changes in the density and size of renal lysosomes appear to be associated with the exocytosis of lysosomal enzymes by hPTH(1-34). PMID- 10467164 TI - Human subtilisin-like proprotein convertase, PACE4 (SPC4) gene expression is highly regulated through E-box elements in HepG2 and GH4C1 cells. AB - PACE4 (SPC4) is a member of the mammalian subtilisin-like proprotein convertase (SPC) family, which participates in maturation of precursor proteins. PACE4 is expressed at high levels in the anterior pituitary, central nervous system, the developing olfactory bulb, heart, and liver. Recently, we determined the gene structure of human PACE4. [Tsuji et al. (1997) J. Biochem. 122, 438-452]. The 5' flanking region of PACE4 gene contains 12 E-boxes (E1 to E12) within 1 kb upstream of the transcription initiation site. To examine the function of these E box elements in the regulation of PACE4 expression, deletion and mutation constructs of the 5'-flanking region were ligated to the luciferase gene and analyzed for promoter activity in HepG2 and GH4C1 cells, which express PACE4 at high level. Some differences were observed in the activity of each promoter construct between HepG2 and GH4C1 cells, although the overall profiles of activity for the promoter fragment series were similar regardless of cell type. We showed that the basal promoter activity of the PACE4 gene is first determined by sequences lying between -315 and -1 bp and further regulated by positive and negative elements in the upstream region. Site-directed mutagenesis of E-boxes in these regulatory elements showed that the E10 E-box act as positive regulator, whereas an E-box cluster (E4-E9) acts as a negative regulator in both cells. E2 E box acts as a positive regulator only in HepG2 cells. Other E-boxes (E1, E3, and E12) had no effect on the promoter activity. These results indicate that E-box elements play a critical role in controlling PACE4 expression in HepG2 and GH4C1 cells and that PACE4 expression is regulated by a mechanism distinct from that of other SPC family proteases. PMID- 10467165 TI - Histamine monoclonal antibody for brain immunocytochemistry. AB - Among five monoclonal antibodies (AHA-1 to 5 mAbs) prepared against glutaraldehyde (GA)-conjugated histamine (HA) in our previous study, only mAb AHA 2 was found to detect HA specifically in rat brain neurons by an immunocytochemistry method (ICC) using GA as a tissue fixative. All the other mAbs, except for AHA-5, reacted with HA in the enterochromaffin-like cells (ECL cells) of rat stomach [Fujiwara et al. (1997) Histochem. Cell Biol. 107, 39-45]. Enzyme-linked immunosorbent assay (ELISA) binding and inhibition tests demonstrated that AHA-2 is specific for HA, with almost no detectable cross reaction with any other established or putative amino acid neurotransmitters, LH RH, TRH, or peptides with N-terminal histidines. ELISA assays also suggested that the AHA-2 mAb recognizes a HA epitope structure different from the one recognized by the AHA-1 mAb. The immunostaining patterns with AHA-2 mAb, as seen in the five subgroups of the tuberomammillary nuclei in the rat posterior hypothalamus, were very similar to those described by Inagaki et al. [(1988) Brain Res. 439, 402 405; (1990) Exp. Brain Res. 80, 374-380] and Panula et al. [(1984) Proc. Natl. Acad. Sci. USA 81, 2572-2576; (1988) J. Histochem. Cytochem. 36, 259-269] using polyclonal anti-HA serum. However, it was also noted that moderate numbers of immunoreactive nerve fibers projected into the median eminence. The present HA ICC method using AHA-2 mAb allows highly sensitive HA detection in brain, and thus might permit detailed studies of HA localization hitherto impossible using previously available anti-HA polyclonal antibodies produced against carbodiimide conjugated HA. PMID- 10467166 TI - Azide- and cyanide-binding to the Escherichia coli bd-type ubiquinol oxidase studied by visible absorption, EPR and FTIR spectroscopies. AB - Cytochrome bd-type ubiquinol oxidase contains two hemes b (b(558) and b(595)) and one heme d as the redox metal centers. To clarify the structure of the reaction center, we analyzed Escherichia coli cytochrome bd by visible absorption, EPR and FTIR spectroscopies using azide and cyanide as monitoring probes for the exogenous ligand binding site. Azide-binding caused the appearance of a new EPR low-spin signal characteristic of ferric iron-chlorin-azide species and a new visible absorption band at 647 nm. However, the bound azide ((14)N(3)) anti symmetric stretching infrared band (2, 010.5 cm(-1)) showed anomalies upon (15)N substitutions, indicating interactions with surrounding protein residues or heme b(595) in close proximity. The spectral changes upon cyanide-binding in the visible region were typical of those observed for ferric iron-chlorin species with diol substituents in macrocycles. However, we found no indication of a low spin EPR signal corresponding to the ferric iron-chlorin-cyanide complexes. Instead, derivative-shaped signals at g = 3.19 and g = 7.15, which could arise from the heme d(Fe(3+))-CN-heme b(595)(Fe(3+)) moiety, were observed. Further, after the addition of cyanide, a part of ferric heme d showed the rhombic high spin signal that coexisted with the g(z) = 2.85 signal ascribed to the minor heme b(595)-CN species. This indicates strong steric hindrance of cyanide-binding to ferric heme d with the bound cyanide at ferric heme b(595). PMID- 10467167 TI - Genetic analysis of the peptide synthetase genes for a cyclic heptapeptide microcystin in Microcystis spp. AB - Peptide-synthetase-encoding DNA fragments were isolated by a PCR-based approach from the chromosome of Microcystis aeruginosa K-139, which produces cyclic heptapeptides, 7-desmethylmicrocystin-LR and 3,7-didesmethylmicrocystin-LR. Three open reading frames (mcyA, mcyB, mcyC) encoding microcystin synthetases were identified in the gene cluster. Sequence analysis indicated that McyA (315 kDa) consists of two modules with an N-methylation domain attached to the first and an epimerization domain attached to the second; McyB (242 kDa) has two modules, and McyC (147 kDa) contains one module with a putative C-terminal thioesterase domain. Conserved amino acid sequence motifs for ATP binding, ATP hydrolysis, adenylate formation, and 4'-phosphopantetheine attachment were identified by sequence comparison with authentic peptide synthetase. Insertion mutations in mcyA, generated by homologous recombination, abolished the production of both microcystins in M. aeruginosa K-139. Primer extension analysis demonstrated light dependent mcy expression. Southern hybridization and partial DNA sequencing analyses of six microcystin-producing and two non-producing Microcystis strains suggested that the microcystin-producing strains contain the mcy gene and the non producing strains can be divided into two groups, those possessing no mcy genes and those with mcy genes. PMID- 10467168 TI - Multi-specificity of a Psathyrella velutina mushroom lectin: heparin/pectin binding occurs at a site different from the N-acetylglucosamine/N acetylneuraminic acid-specific site. AB - An N-acetylglucosamine (GlcNAc)/N-acetylneuraminic acid-specific lectin from the fruiting body of Psathyrella velutina (PVL) is a useful probe for the detection and fractionation of specific carbohydrates. In this study, PVL was found to exhibit multispecificity to acidic polysaccharides and sulfatides. Purified PVL and a counterpart lectin to PVL in the mycelium interact with heparin neoproteoglycans, as detected by both membrane analysis and solid phase assay. The pH-dependencies of the binding to heparin and GlcNAc5-6 differ. The heparin binding of PVL is inhibited best by pectin, polygalacturonic acid, and highly sulfated polysaccharides, but not by GlcNAc, colominic acid, or other glycosaminoglycans. Sandwich affinity chromatography indicated that PVL can simultaneously interact with heparin- and GlcNAc-containing macromolecules. Extensive biotinylation was found to suppress the binding activity to heparin while the GlcNAc binding activity is retained. On the other hand, biotinyl PVL binds to sulfatide and the binding is not inhibited by GlcNAc, N-acetylneuraminic acid, or heparin. These results indicate that PVL is a multi-ligand adhesive lectin that can interact with various glycoconjugates. This multispecificity needs to be recognized when using PVL as a sugar-specific probe to avoid misleading information about the nature of glycoforms. PMID- 10467169 TI - Host cell proteins bind specifically to the capsid-cleaved 5' end of Leishmaniavirus RNA. AB - Leishmaniavirus (LRV) is a double-stranded RNA (dsRNA) virus that persistently infects some strains of the protozoan parasite, Leishmania. LRV generates a short transcript, corresponding to the 5' end of the positive-sense RNA (320 nt), via a cleavage event mediated by the viral capsid protein on the full-length positive sense RNA transcript. To address the possibility that the RNA cleavage represents a regulatory mechanism for maintaining persistent infection, the interactions between Leishmania cytoplasmic proteins and in vitro synthesized viral transcripts were studied. In gel mobility shift experiments, three specific RNA/protein complexes were formed between cellular proteins and the cleaved viral transcript, and three major proteins were labeled by UV cross-linking. No protein binding activity was observed for either the short (320 nt) or full-length RNA transcripts. However, the two cleavage reaction products were able to form stable RNA/RNA complexes. We present a model in which the virus is targeting its own transcript for cleavage to promote binding of host factors to cryptic domains inaccessible in the full-length transcript. PMID- 10467170 TI - beta-glycosidase from the hyperthermophilic archaeon Sulfolobus solfataricus: structure and activity in the presence of alcohols. AB - beta-Glycosidase from the extreme thermophilic archaeon Sulfolobus solfataricus is a tetrameric protein with a molecular mass of 240 kDa, stable in the presence of detergents, and with a maximal activity at temperatures above 95 degrees C. Understanding the structure-activity relationships of the enzyme under different conditions is of fundamental importance for both theoretical and applicative purposes. In this paper we report the effect of methanol, ethanol, 1-propanol, and 1-butanol on the activity of S. solfataricus beta-glycosidase expressed in Escherichia coli. The alcohols stimulated the enzyme activity, with 1-butanol producing its maximum effect at a lower concentration than the other alcohols. The structure of the enzyme was studied in the presence of 1-butanol by circular dichroism, and Fourier-transform infrared and fluorescence spectroscopies. Circular dichroism and steady-state fluorescence measurements revealed that at low temperatures the presence of the alcohol produced no significant changes in the tertiary structure of the enzyme. However, time-resolved fluorescence data showed that the alcohol modifies the protein microenvironment, leading to a more flexible enzyme structure, which is probably responsible for the enhanced enzymatic activity. PMID- 10467171 TI - Interleukin-1beta induces interleukin-6 production through the production of prostaglandin E(2) in human osteoblasts, MG-63 cells. AB - This study was conducted to investigate the mechanism of interleukin-1beta (IL 1beta)-induced IL-6 production in human osteoblasts (MG-63 cells). Stimulation with IL-1beta resulted in the production of IL-6 and prostaglandin E(2) (PGE(2)). IL-6 production gradually increased and peaked 96 h after stimulation. IL-6 mRNA was detected between 4 and 72 h after IL-1beta stimulation. The patterns of PGE(2) production and the expression of cyclooxygenase-2 (COX-2) mRNA were biphasic after stimulation. Actinomycin D, cycloheximide, indomethacin, and NS 398 (COX-2 inhibitor) suppressed the production of IL-6 and PGE(2). Anti-PGE(2) antibody markedly reduced the production of IL-6. In addition, stimulation with 17-phenyl-PGE(2), a PGE receptor-1 (EP-1 receptor) agonist, led to the expression of IL-6 mRNA after pretreatment with IL-1beta. These findings indicate that IL 1beta-induced IL-6 production in MG-63 cells involves the following sequence of steps: IL-1beta-induced COX-2 activation, PGE(2) production, and EP-1 receptor signaling prior to IL-6 production. PMID- 10467172 TI - Crystal structure of prolyl aminopeptidase from Serratia marcescens. AB - Prolyl aminopeptidase from Serratia marcescens specifically catalyzes the removal of N-terminal proline residues from peptides. We have solved its three dimensional structure at 2.3 A resolution by the multiple isomorphous replacement method. The enzyme consists of two contiguous domains. The larger domain shows the general topology of the alpha/beta hydrolase fold, with a central eight stranded beta-sheet and six helices. The smaller domain consists of six helices. The catalytic triad (Ser113, His296, and Asp268) is located near the large cavity at the interface between the two domains. Cys271, which is sensitive to SH reagents, is located near the catalytic residues, in spite of the fact that the enzyme is a serine peptidase. The specific residues which make up the hydrophobic pocket line the smaller domain, and the specificity of the exo-type enzyme originates from this smaller domain, which blocks the N-terminal of P1 proline. PMID- 10467173 TI - Protein design of geranyl diphosphate synthase. Structural features that define the product specificities of prenyltransferases. AB - Geranyl diphosphate synthase catalyzes the condensation of isopentenyl diphosphate with dimethylallyl diphosphate to give a C(10) compound, geranyl diphosphate, which is a precursor of all monoterpenoids. However, the gene has not been isolated from any organisms. To examine the possibility that geranyl diphosphate synthase has evolved from a common ancestor of the prenyltransferase family and to predict the active site structure, we tried to convert Bacillus stearothermophilus farnesyl diphosphate synthase to geranyl diphosphate synthase, according to our previous findings. Several mutated farnesyl diphosphate synthases that have single amino acid substitutions before the first aspartate rich motif were constructed. A mutated enzyme that has the replacement of serine by phenylalanine at the fourth position before the motif exclusively produced geranyl diphosphate when dimethylallyl diphosphate was used as the primer, and hardly accepted geranyl diphosphate as a primer, indicating that this mutation causes the conversion to geranyl diphosphate synthase. This result supports the idea that the product specificities of all members of the E-prenyltransferase family are mainly defined by a few structural features: the amino acids at the fourth position and the fifth position before the first aspartate-rich motif, and the insertion of two amino acids in the motif. This suggests that natural geranyl diphosphate synthases might have an active site structure similar to that of the mutated enzyme. PMID- 10467174 TI - Genomic structure of the amphioxus calcium vector protein. AB - Calcium vector protein (CaVP) is an EF-hand Ca(2+)-binding protein, which is unique to the protochordate, amphioxus. CaVP is supposed to act as a Ca(2+) signal transductor, but its exact function remains unknown. Not only its function but also its exact evolutionary relationship to other Ca(2+)-binding proteins is unclear. To investigate the evolution of CaVP, we have determined the complete sequences of CaVP cDNAs from two amphioxus species, Branchiostoma lanceolatum and B. floridae, whose open reading frame cDNA and amino acid sequences show 96.5 and 98.2% identity, respectively. We have also elucidated the structure of the gene of B. floridae CaVP, which is made up of seven exons and six introns. The positions of four of the six introns (introns 1, 2, 3, and 5) are identical with those of calmodulin, troponin C, and the Spec protein of the sea urchin. These latter proteins belong to the so-called troponin C superfamily (TnC superfamily) and thus CaVP likely also belongs to this family. Intron 6 is positioned in the 3' noncoding region and is unique to CaVP, so it may represent a landmark of the CaVP lineage only. The position of intron 4 is not conserved in the genes of the TnC superfamily or CaVP, and seems to result from either intron sliding or the addition of an intron (randomly inserted into or close to domain III) to the genes of the TnC superfamily during their evolution. PMID- 10467175 TI - Phosphocarrier proteins in an intracellular symbiotic bacterium of aphids. AB - A GroEL homolog produced by Buchnera, an intracellular symbiotic bacterium of aphids, is not only a molecular chaperone but also a novel phosphocarrier protein, suggesting that this protein plays a role in a signal transducing system specific to bacteria living in an intracellular environment. This prompted us to look into phosphocarrier proteins of Buchnera that may be shared in common with other bacteria. As a result, no evidence was obtained for the presence of sensor kinases of the two-component system in Buchnera, which are found in many bacteria. It is possible that the lack of sensor kinases is compensated for by the mulitifunctional GroEL homolog in this symbiotic bacteria. In contrast, we successfully identified three phosphotransferase system genes, ptsH, ptsI, and crr in Buchnera, and provide evidence for their active expression. While the deduced amino acid sequences of these gene products, histidine-containing phosphocarrier protein, Enzyme I, and Enzyme III were similar to their counterparts in Escherichia coli, the predicted isoelectric points of the Buchnera proteins were strikingly higher. It was also suggested that Buchnera Enzyme I, when produced in E. coli, is able to accept the phosphoryl group from phosphoenolpyruvate, but not from ATP. PMID- 10467176 TI - Construction of a screening system for selecting lysozyme mutants unable to form a stable structure from random mutants. AB - To collect folding information, we screened and analyzed the recombinant hen lysozyme mutants which were not secreted from yeast. As model mutants, Leu8Arg, Ala10Gly, and Met12Arg were prepared by site-directed mutagenesis and analyzed as to whether they were secreted from yeast or not. Consequently, Ala10Gly was found to be secreted from yeast, but Leu8Arg and Met12Arg were not. Next, these mutants were expressed in Escherichia coli and refolded in vitro. As a result, Ala10Gly folded as the wild-type did. Leu8Arg efficiently refolded in renaturation buffer containing glycerol. Met12Arg did not refold even in the presence of glycerol. These results show that the Ala10Gly mutation does not affect folding or stability, that Leu8Arg is too unstable to be secreted from yeast, and that Met12Arg may be very unstable or the mutation affects the folding pathway. We screened the mutants that were not secreted by yeast from a randomly mutated lysozyme library, and obtained Asp18His/Leu25Arg and Ala42Val/Ser50Ile/Leu56Gln. These two mutants were expressed in E. coli and then refolded in the presence of urea or glycerol. These mutants were refolded only in the presence of glycerol. Each single mutant of Asp18His/Leu25Arg and Ala42Val/Ser50Ile/Leu56Gln was independently prepared and folded in vitro. The results showed that Leu25Arg and Leu56Gln were the dominant mutations, respectively, which cause destabilization. These results show that the mutant lysozymes which were not secreted from yeast may be unstable or have a defect in the folding pathway. Thus, we established a screening system for selecting mutants which are unable to form a stable structure from random mutants. PMID- 10467178 TI - Preparation of fluorescence-labeled GM1 and sphingomyelin by the reverse hydrolysis reaction of sphingolipid ceramide N-deacylase as substrates for assay of sphingolipid-degrading enzymes and for detection of sphingolipid-binding proteins. AB - Sphingolipid ceramide N-deacylase is an enzyme capable of hydrolyzing the N-acyl linkages of ceramides of various sphingolipids. Recently, it was found that the enzyme catalyzes the reverse hydrolysis reaction in which free fatty acids are condensed to lyso-sphingolipids to produce sphingolipids. This paper describes a simple method for the synthesis of fluorescence-labeled sphingolipids utilizing the condensation reaction of the enzyme. N-TFAc-aminododecanoic acids were efficiently condensed by the enzyme to the lyso-forms of GM1 and sphingomyelin in glycine buffer (pH 10). The reaction products, N-TFAc-amino-GM1 and sphingomyelin, were obtained with overall yields of 60%. The purified products were identified to be omega-amino-GM1 and omega-amino-sphingomyelin, respectively, by TLC and FAB-MS or ESI-LC/MS analysis after removal of the N-TFAc by mild alkaline treatment. NBD-labeled GM1 and sphingomyelin were prepared from omega-amino-GM1 and omega-amino-sphingomyelin by coupling with 4-fluoro-NBD. These fluorescence-labeled substrates, C12-NBD-GM1 and C12-NBD-sphingomyelin, were hydrolyzed by endoglycoceramidase and sphingomyelinase, respectively, to produce NBD-dodecanoylsphingosines, but were resistant to hydrolysis by sphingolipid ceramide N-deacylase. C12-NBD-sphingomyelin was found to be a better substrate than the commercially available C6-NBD-sphingomyelin for the assay of sphingomyelinase from various sources. We also describe a new method to detect GM1-binding proteins using fluorescence-labeled GM1. PMID- 10467177 TI - Inactivation of proprotein convertase, PACE4, by alpha1-antitrypsin Portland (alpha1-PDX), a blocker of proteolytic activation of bone morphogenetic protein during embryogenesis: evidence that PACE4 is able to form an SDS-stable acyl intermediate with alpha1-PDX. AB - PACE4 (SPC4), a member of the subtilisin-like proprotein convertase (SPC) family of proteases that cleave at paired basic amino acids, exhibits a dynamic expression pattern during embryogenesis and colocalizes with bone morphogenetic proteins (BMPs). Recently Cui et al. reported that the ectopic expression of alpha1-antitrypsin variant Portland (alpha1-PDX), an engineered serpin that contains the minimal SPC consensus motif in its reactive loop, blocks the proteolytic activation of BMP4, leading to abnormal embryogenic development [Cui, Y. et al. (1998) EMBO J. 17, 4735-4743]. TGFbeta-related factors such as BMPs are synthesized as inactive precursors and activated by limited proteolysis at multibasic amino acids. Therefore, an alpha1-PDX-inhibitable protease is thought to participate in BMP activation. However, conflicting properties, including sensitivity to alpha1-PDX, have been reported for PACE4. In this study, we examined whether alpha1-PDX is responsible for the inhibition of PACE4 by measuring the protease/inhibitor complex directly. Here we show that alpha1-PDX has the ability to form an SDS-stable acyl-intermediate (180 kDa) with PACE4 in vivo and in vitro. Further, we characterized the PACE4 secreted into the culture medium from Cos-1 cells by a specific immunological assay. An alpha1-PDX insensitive and decanoyl-RVKR-chloromethylketone-sensitive 60-kDa protease(s) is greatly activated in conditioned medium by PACE4 overexpression, suggesting that the activation of an unknown protease(s) other than PACE4 is the cause of the variation in the properties of PACE4. PACE4 is a Ca(2+)-dependent protease with an optimal Ca(2+) requirement of 2 mM, and shows its highest activity at weakly basic pH. PACE4 activity is completely inhibited by EDTA and EGTA, but not by leupeptin. These results show that PACE4 activity can be inhibited by alpha1-PDX as well as furin (SPC1) and suggest that the inhibition of PACE4-mediated activation of factors such as BMPs by alpha1-PDX causes abnormal embryogenic development. PMID- 10467179 TI - Isolation and characterization of cytosolic and membrane-bound deubiquitinylating enzymes from bovine brain. AB - The deubiquitinylating enzymes (DUBs), that release free ubiquitin (Ub) from its precursors or ubiquitinylated proteins, are known to comprise of a large protein family in eukaryotes, but those in mammalian tissues remain largely unknown. Here we report the existence of unexpectedly large species of DUBs in both soluble and membrane-bound fractions of bovine brain, based on their ability to cleave (125)I labeled Ub-fused alphaNH-MHISPPEPESEEEEEHYC (designated as Ub-PESTc). Two cytosolic enzymes, tentatively called sDUB-1 and sDUB-2, with molecular masses of about 30 kDa were purified to near homogeneity by Ub-Sepharose affinity chromatography. sDUB-1 and sDUB-2 corresponded to UCH-L3 and UCH-L1/PGP 9.5, respectively. Intriguingly, the particulate fraction of the brain homogenate was found to also contain strong activities against (125)I-Ub-PESTc, which can be solubilized by treatment with 5% n-heptyl-beta-D-thioglucoside and 1% Nonidet P 40, but not by washing with 1 M NaCl. From the solubilized material, two new 30 kDa, membranous DUBs (called mDUB-1 and mDUB-2) were purified to apparent homogeneity by Ub-Sepharose chromatography. Two other Ub-aldehyde sensitive DUBs, designated as mDUB-3 and mDUB-4, were also partially purified by conventional chromatographic operations. These mDUBs differed from each other in substrate specificity and exhibited different characteristics from the sDUBs, revealing that they are a new type of membrane-bound DUB. These results indicate the presence of divergent DUBs in mammalian brain, which may contribute to regulation of numerous pivotal cellular functions mediated by the covalent modification of Ub. PMID- 10467180 TI - Steroid monooxygenase of Rhodococcus rhodochrous: sequencing of the genomic DNA, and hyperexpression, purification, and characterization of the recombinant enzyme. AB - Steroid monooxygenase of Rhodococcus rhodochrous is a Baeyer-Villigerase catalyzing the insertion of an oxygen atom between the C(17)- and C(20)-carbons of progesterone to produce testosterone acetate. The 5.1-kbp-long BamHI DNA fragment containing the steroid monooxygenase gene, smo, was cloned from the chromosomal DNA and sequenced. The smo gene is 1,650 nucleotides long, starts with a TTG codon, and ends with a TGA codon. The deduced amino acid sequence indicates that the enzyme protein consist of 549 amino acid residues with a molecular mass of 60,133. Thus, the molecular mass of the holoenzyme is 60,919. The amino acid sequence is highly homologous (41.2% identity) to that of cyclohexanone monooxygenase of Acinetobacter sp. In the upstream of the smo gene, the genes of heat shock proteins, dnaK, grpE, and dnaJ, located on the complementary strand, and the DNA-inserts of pSMO and pD1, which contains the ksdD gene, were joined at the BamHI site of the dnaJ gene. The smo gene was modified at the initiation codon to ATG and ligated with an expression vector to construct a plasmid, pSMO-EX, and introduced into Escherichia coli cells. The transformed cells hyperexpressed the steroid monooxygenase as an active and soluble protein at more than 40 times the level in R. rhodochrous cells. Purification of the recombinant monooxygenase from the E. coli cells by simplified procedures yielded about 2.3 mg of enzyme protein/g wet cells. The purified recombinant steroid monooxygenase exhibited indistinguishable molecular and catalytic properties from those of the R. rhodochrous enzyme. PMID- 10467181 TI - Interconversion of Mn(2+)-dependent and -independent protein phosphatase 2A from human erythrocytes: role of Zn(2+) and Fe(2+) in protein phosphatase 2A. AB - Human erythrocyte Mn(2+)-dependent (C'A') and -independent (CA) protein serine/threonine phosphatase (PP) 2A are composed of 34-kDa catalytic C' and C subunits, in which the metal dependency resides, and 63-kDa regulatory A' and A subunits, respectively. Each catalytic and regulatory subunit gave the same V8- and papain-peptide maps, respectively. Stoichiometric zinc and substoichiometric iron were detected in CA but not in C'A' [Nishito et al. (1999) FEBS Lett. 447, 29-33]. The Mn(2+)-dependent protein-tyrosine phosphatase (PTP) activity of C'A' was about 70-fold higher than that of CA. Pre-incubation of CA with 25 mM NaF changed CA to a Mn(2+)-dependent form with higher PTP activity. The same NaF treatment had no effect on C'A'. Pre-incubation of C'A' with ZnCl(2), zinc metallothionein, or FeCl(2) activated the Mn(2+)-independent PP activity, but pre incubation with FeCl(3) did not. Ascorbate in the pre-incubation and assay mixture significantly stimulated the effect of FeCl(2). Pre-incubation of C'A' with 5 microM ZnCl(2) and 15 microM FeCl(2) in the presence of 1 mM ascorbate synergistically stimulated the Mn(2+)-independent PP activity, with concomitant suppression of the Mn(2+)-dependent PP and PTP activities. The PP and PTP activities of CA were unaffected by the same zinc and/or iron treatment. Micromolar concentrations of vanadate strongly inhibited the Mn(2+)-dependent PP activity of C'A' but only slightly inhibited the PP activity of CA. Using the distinct effect of vanadate as an indicator, the interconversion between CA and C'A' with the above mentioned treatments was proved. These results support the notion that Mn(2+)-independent CA is a Zn(2+)- and Fe(2+)-metalloenzyme, whose apoenzyme is Mn(2+)-dependent C'A'. PMID- 10467182 TI - Beta-blocker withdrawal: the song of Orpheus. PMID- 10467183 TI - To train or not to train. PMID- 10467184 TI - Transradial cardiac catheterization: is femoral access obsolete? PMID- 10467185 TI - Practice still makes perfect. PMID- 10467186 TI - Racial differences in the use of invasive cardiac procedures: A continuous quality improvement approach. PMID- 10467187 TI - Long-term nitrate use in chronic coronary artery disease: need for a randomized controlled trial. PMID- 10467188 TI - Clinical features of Wolff-Parkinson-White syndrome. AB - Wolff-Parkinson-White syndrome is the most common form of ventricular preexcitation. Understanding this syndrome is fundamental for anyone interested in learning about arrhythmias. This review addresses (1) the historic sequence of events that led to the understanding of this syndrome; (2) the pathologic, embryologic, and electrophysiologic properties of accessory pathways; (3) the epidemiology and genetics of this syndrome; (4) the clinical diagnosis of this syndrome, with special emphasis on the arrhythmias that patients with ventricular preexcitation are predisposed to; and (5) the therapy for patients with Wolff Parkinson-White syndrome. PMID- 10467189 TI - Assessing the cost of a cardiology residency program with a cost construction model. AB - BACKGROUND: Although the total costs of graduate medical education are difficult to quantify, this information is of great importance in planning over the next decade. METHODS AND RESULTS: A cost construction model was used to quantify the costs of teaching faculty, cardiology fellows' salaries and benefits, overhead (physical plant, equipment, and support staff), and other costs associated with the cardiology residency program at the University of Texas-Houston during the 1996 to 1997 academic year. Surveys of cardiology faculty and fellows, checked by the program director, were conducted to determine the time spent in teaching activities; access to institutional and departmental financial records was obtained to quantify associated costs. The model was then developed and examined for a range of assumptions concerning cardiology fellows' productivity, replacement costs, and the cost allocation of activities jointly producing clinical care and education. The instructional cost of training (cost of didactic, direct clinical supervision, preparation for teaching, and teaching related administration, plus the support of the teaching program) was estimated at $73,939 per cardiology fellow per year. This cost was less than the estimated replacement value of the teaching and clinical services provided by cardiology fellows, $100,937 per cardiology fellow per year. Sensitivity analysis, with different assumptions on cardiology fellows' productivity and replacement costs, varied the cost estimates but generally represented the cardiology residency program as an asset. CONCLUSIONS: Cost construction models can be used as a tool to estimate variations in resource requirements resulting from changes in curriculum or educators' costs. In this residency, the value of the teaching and clinical services provided by cardiology fellows exceeded the cost of the resources used in the educational program. PMID- 10467190 TI - Predictors of patient-reported physical and mental health 6 months after percutaneous coronary revascularization. AB - BACKGROUND: There is interest in measuring and comparing outcomes of percutaneous transluminal coronary angioplasty (PTCA) other than death, but there are no accepted methods for adjusting these outcomes for preprocedure differences in populations. We sought to identify independent predictors of functional outcome after PTCA. METHODS AND RESULTS: We developed multivariate risk adjustment models for the 6-month postprocedure physical and mental health summary scores of the MOS SF-36. Complete data were available on 1182 patients undergoing PTCA at 12 institutions. The mean physical component score (PCS) of the SF-36 rose from 36.6 before PTCA to 43. 4 at 6 months after PTCA (P <.0001). Independent predictors of follow-up PCS were baseline PCS, a composite index of comorbidities, prior coronary bypass surgery, baseline MOS SF-36 mental component score (MCS), age, and recent thrombolysis. The model had an adjusted R(2) value of 0.357. The mean MCS rose from 48.5 before PTCA to 50.5 at 6 months after PTCA (P <.0001). Independent predictors of postprocedure mental health were baseline MCS, age, and heart failure. The predictive model for MCS had an adjusted R(2) value of 0.235. CONCLUSIONS: Preprocedure patient-reported functional status and select clinical variables are significantly associated with physical functioning and mental health 6 months after PTCA. The predictive power of these models, however, is probably insufficient to allow their use for comparisons among institutions or providers. PMID- 10467191 TI - Effect of transradial access on quality of life and cost of cardiac catheterization: A randomized comparison. AB - BACKGROUND: Transradial access is a recently developed alternative for diagnostic cardiac catheterization. Its effects on quality of life after the procedure, patient preference, and cost are unknown. METHODS AND RESULTS: We performed a randomized single-center trial in which 99 patients underwent transfemoral and 101 underwent transradial diagnostic cardiac catheterization. Quality of life was measured with the SF-36 and visual analog scales at baseline, 1 day, and 1 week. Patients were examined at 1 day and at 1 week after for complications. Costs were measured prospectively. One patient in the femoral group and 2 in the radial group crossed over to the alternative access site. There were no major access site complications. One patient in the transfemoral group had a minor stroke. Transradial catheterization significantly reduced median length of stay (3.6 vs 10.4 hours, P <.0001). Over the first day after the procedure, measures of bodily pain, back pain, and walking ability favored the transradial group (P <.05 for all comparisons). Over the week after the procedure, changes in role limitations caused by physical health, bodily pain, and back pain favored the transradial group (P <.05 for all comparisons). There was a strong patient preference for transradial catheterization as well (P <. 0001). Transradial catheterization led to significant reductions in bed, pharmacy, and total hospital costs ($2010 vs $2299, P <.0001). CONCLUSIONS: Among patients undergoing diagnostic cardiac catheterization, transradial access leads to improved quality of life after the procedure, is strongly preferred by patients, and reduces hospital costs. PMID- 10467192 TI - Coronary artery stent outcomes in a Medicare population: less emergency bypass surgery and lower mortality rates in patients with stents. AB - BACKGROUND: Randomized trials of coronary stents versus conventional balloon angioplasty have demonstrated improved short- and long-term outcomes for selected patients receiving stents. The purpose of this study was to compare outcomes in patients receiving stents with those undergoing conventional balloon angioplasty in everyday clinical practice. METHODS AND RESULTS: This study uses information from the Medicare Provider Analysis and Review files for fiscal years 1994 and 1996, the first year the coronary stent code was used. For patients 65 years of age and older, 165,657 cases in 1994 and 201,869 in 1996, including 74,836 cases with stent placement, were identified. Outcomes included hospital deaths, use of same- admission coronary artery bypass surgery, and either or both. Analyses were performed separately for those with and those without a principal diagnosis of acute myocardial infarction. Hospital mortality rates were similar in both years, but the use of same-admission coronary artery bypass surgery was lower in 1996. In that year, for both patients with and those without acute myocardial infarction, hospital death and the use of same-admission coronary artery bypass surgery were lower in the stent group. Additionally, results in the stent group were generally better at high-volume (>200 cases per year) institutions, as was the case for the prestent, 1994 results. CONCLUSIONS: This study documents improved short-term outcomes in older patients who undergo coronary stent placement. Stenting did not eliminate the finding of improved outcomes at high volume centers. PMID- 10467193 TI - Long-term outcome of patients with very long stents for treatment of diffuse coronary disease. AB - OBJECTIVES: The study sought to determine the 6-month clinical outcome of patients who underwent implantation of very long coronary stents to treat diffuse disease and/or long dissections and to compare the findings with those reported in the literature for patients who underwent implantation of multiple short coronary stents. BACKGROUND: New designs of flexible stents enable the implantation of long stents rather than multiple short, older design stents. The initial experience is very promising but the long-term outcome has not been described yet. METHODS: Fifty-seven consecutive patients in whom 67 long stents (>/=30 mm) were successfully deployed were included in this study. Six-month clinical and angiographic follow-up was prospectively collected. Patients with recurrent angina underwent coronary angiography without further testing. Patients who remained asymptomatic at the 6-month follow-up visit underwent positron emission tomographic imaging, and those with results suggestive of ischemia underwent coronary angiography. A combined study end point was defined as death, myocardial infarction, and the need for target vessel revascularization. RESULTS: Only 1 patient (2%) reached a study end point at hospital discharge. An additional 20 patients (total 21 patients [37%]) reached an end point by 6 months. The outcome was not influenced by the clinical presentation (stable or unstable angina) or by the indication for stenting (elective or emergency). Predictors for adverse outcome were multiple stents per narrowing (63% vs 29%, P <. 04), and stents smaller than 3.5 mm (49% vs 22%). Narrowing and stent length were not predictive of a study end point in narrowings that were successfully treated by a single long stent. CONCLUSIONS: Elective stenting provides an effective solution for patients with diffuse coronary disease provided that a single long stent (usually <40 mm) can cover the full length of the narrowing. The results are better when vessels larger than 3 mm are treated. Compared with multiple short stents, implantation of a single long stent is probably at least as effective, and the procedure is quicker and cheaper and thus should be the preferred approach. PMID- 10467194 TI - Primary stenting in acute myocardial infarction: influence of diabetes mellitus in angiographic results and clinical outcome. AB - BACKGROUND: The outcome of patients with diabetes after myocardial infarction (MI) has traditionally been worse than in their nondiabetic counterparts before and during the thrombolytic therapy era. Whether the fate of patients with diabetes might improve with mechanical intervention, particularly with primary stenting, has not previously been studied. METHODS: We compared the angiographic and clinical outcome of 76 nondiabetic patients (aged 61 +/- 14 years; 66% male) and 28 patients with diabetes (aged 65 +/- 12 years; 64% male) consecutively treated with primary stenting for acute MI. Coronary Thrombolysis In Myocardial Infarction grade 3 flow was restored in 96% of diabetic and 97% of nondiabetic patients. RESULTS: Angiographic results after stent deployment were similar in the 2 groups. At 1-month follow-up, all patients in both groups were alive. Patients with diabetes had a much higher incidence of stent thrombosis (18% vs 1%; P =.003), which accounted for the majority of the major cardiac events at 1 month (21% vs 4%; P =.009). At a mean follow-up of 315 +/- 13 days, 99% of nondiabetic and 89% of patients with diabetes were alive (P =.04). Overall freedom from a major cardiac event (death, MI, target vessel revascularization) at 315 +/- 13 day follow-up was 88% for nondiabetics and 54% for patients with diabetes (P =.0003). By multivariate analysis, diabetes mellitus was the most important predictor for development of 1-month (RR 9.89; 95% confidence interval, 1.6-30) and late major cardiovascular events (RR 8.39; 95% confidence interval, 2.93-24). CONCLUSIONS: Primary stenting in acute MI is highly effective in restoring immediate TIMI 3 coronary flow in nondiabetic patients and patients with diabetes. This procedure may improve benefit in terms of mortality rate to both groups, particularly in patients with diabetes, compared with previous reports with thrombolytic therapy. Nevertheless, stent thrombosis and major cardiovascular events at 1 month and late follow-up are more frequent in patients with diabetes. PMID- 10467195 TI - Can beta-blocker therapy be withdrawn from patients with dilated cardiomyopathy? AB - BACKGROUND: It has yet to be determined whether withdrawing beta-blocker therapy from patients with dilated cardiomyopathy (DCM) is safe. METHODS: The influence of tapering and then stopping metoprolol was clarified in 13 patients with DCM who had been receiving this agent for >/=30 months. RESULTS: Seven of the 13 patients deteriorated, including 4 who died suddenly or of congestive heart failure during the 4-month period after metoprolol cessation. CONCLUSION: In patients with DCM in whom beta-blocker therapy is effective, withdrawal of these agents may lead to death. We conclude that beta-blockers should not be stopped in this patient group. PMID- 10467196 TI - Pulmonary function, cardiac function, and exercise capacity in a follow-up of patients with congestive heart failure treated with carvedilol. AB - BACKGROUND: Chronic heart failure causes disturbances in ventilation and pulmonary gas transfer that participate in limiting peak exercise oxygen uptake (VO(2p )). The beta-adrenergic receptor blocker carvedilol improves left ventricular (LV) function and not VO(2p). This study was aimed at investigating the pulmonary response to changes in LV performance produced by carvedilol in patients with chronic heart failure. METHODS: Twenty-one patients with New York Heart Association class II to III heart failure were randomly assigned (2 to 1) to carvedilol (25 mg twice daily, n = 14) or placebo (n = 7) for 6 months. Rest forced expiratory volume (FEV(1)), vital capacity, total lung capacity, carbon monoxide diffusing capacity, its alveolar-capillary membrane component, pulmonary venous and transmitral flows (for monitoring changes in LV end-diastolic pressure), LV diastolic and systolic dimensions, stroke volume, ejection fraction, and fiber shortening velocity were measured at baseline and at 3 and 6 months. VO(2p), peak ratio of dead space to tidal volume (VD/VT(p)), ventilatory equivalent for carbon dioxide production (VE/VCO(2)), and VO(2) at anaerobic threshold (VO(2at)) were also determined. RESULTS: FEV(1), vital capacity, total lung capacity, carbon monoxide diffusing capacity, and the alveolar-capillary membrane component were impaired in chronic heart failure compared with 14 volunteers and did not vary with treatment. Carvedilol reduced end-diastolic pressure, end-diastolic diameter, and end-systolic diameter and increased ejection fraction, stroke volume, and fiber shortening velocity without affecting VO(2p), VO(2at), VD/VT(p), or VE/VCO(2) at 3 and 6 months. Placebo did not produce significant changes. CONCLUSIONS: In chronic heart failure carvedilol ameliorates LV function at rest and does not significantly affect ventilation and pulmonary gas transfer or functional capacity. These results suggest that improvement in cardiac hemodynamics with carvedilol does not reverse pulmonary dysfunction. Persistent lung impairment might have some role in the failure of carvedilol to improve exercise performance. PMID- 10467197 TI - Effect of distal stenosis of internal thoracic artery bypass grafts on longitudinal phasic blood flow velocity characteristics. AB - BACKGROUND: Left internal thoracic artery (LITA) bypass conduits show gradual longitudinal transition in their phasic flow velocity patterns from the proximal to distal segments, but little is known about the influence of distal stenosis, particularly early after surgery, on that characteristic. The purpose of this study was to evaluate the influence of distal stenosis on these flow velocity patterns. METHODS: We examined 24 LITAs within 1 month (7 to 30 days) after surgery with a Doppler-tipped guide wire at the proximal, mid, and distal segments. Maximum peak velocities (MPV), time averaged peak velocities (APV), and velocity-time integrals (VTI) were measured. RESULTS: In LITAs without stenosis (n = 14, group A), the APV, MPV, and VTI values at the diastole were significantly greater than those for distal stenosis (minimal lumen diameter >75%, n = 10, group B). The values of the 3 indexes at the systole in each segment did not differ significantly between the 2 groups. Both groups showed gradual increases in the diastolic/systolic ratios of the 3 indexes from the proximal to distal portions, the ratios in group A being significantly larger than that in group B (APV, P <. 001; MPV, P <.01; TVI, P <.01, respectively). For these indexes, sensitivity and specificity for predicting stenosis of LITA was higher in the proximal and mid portion than in the distal. CONCLUSIONS: Anastomotic stenosis decreases the diastolic flow component but not the systolic one. By using diastolic/systolic ratios of the 3 indexes, it is possible to predict distal stenosis of LITA from the resting phasic flow velocity pattern. PMID- 10467198 TI - Symptoms, left ventricular function, and timing of valve replacement surgery in patients with aortic regurgitation. AB - BACKGROUND: Because cardiac decompensation is subtle, the best time to perform aortic valve replacement surgery may be difficult to determine. We investigated the relation of symptoms to left ventricular (LV) function and the timing of valve replacement in patients with aortic regurgitation (AR) of largely rheumatic origin. METHODS: Sixty-eight initially asymptomatic patients (mean age 29 years) with severe chronic AR were monitored for 36 months. Assessments included baseline and yearly echocardiograms and radioisotope ventriculography (resting and exercise) and clinical examinations every 6 months. RESULTS: Forty-seven patients (69%) remained asymptomatic and 21 (31%) had symptoms develop after 24 to 36 months. Compared with symptomatic patients, asymptomatic patients had significantly (P <.05) lower baseline LV end-diastolic diameter, end-systolic diameter, end-systolic stress, and volume/mass ratio but greater shortening fraction and ejection fraction (EF) at rest. These variables remained stable without statistically significant change until surgical correction in symptomatic patients. Percent variation of EF from rest to exercise increased in patients who remained asymptomatic (EF 2.8% +/- 10.6%) but decreased in those who became symptomatic (EF -4.2% +/- 13%; P <.05). Twenty symptomatic patients (New York Heart Association class III/IV, angina and/or syncope) had valve replacement surgery, after which all were in New York Heart Association class I/II and had significant decreases of LV end-diastolic and end-systolic diameters and an increase on percent variation of EF from rest to exercise (P <.0001). CONCLUSIONS: Development of symptoms did not correlate with change in any ventricular function indexes. Surgery on appearance of symptoms restored LV function to near normal. PMID- 10467199 TI - Anthropometric and physiologic correlates of mitral valve prolapse in a biethnic cohort of young adults: the CARDIA study. AB - OBJECTIVE: To describe the epidemiology of echocardiographic mitral valve prolapse (MVP) and its anthropometric, physiologic, and psychobehavioral correlates with a cross-sectional analysis at 4 urban clinical centers. PATIENTS: A biethnic, community-based sample of 4136 young (aged 23 to 35 years) adult participants in the Coronary Artery Risk Development in Young Adults (CARDIA) study who had echocardiograms during their third examination between 1990 and 1991. MEASUREMENTS: Echocardiographic mitral valve prolapse, Doppler mitral regurgitation, blood pressure, anthropometry, and 4 psychobehavioral scales. RESULTS: Definite echocardiographic MVP prevalence was 0.6% overall and was similar across the 4 ethnicity/sex groups. Most participants (21 of 26, 80%) with definite echocardiographic MVP were unaware of their condition. Relative to persons with normal echocardiograms, those with echocardiographic MVP were taller (174.6 cm vs 171.0 cm, P <.01), leaner (26.7 mm vs 37.4 mm sum of triceps and subscapular skinfolds, P <.01), had lower body mass index (22.0 kg/m(2) vs 26.2 kg/m(2), P <.01), and more often has Doppler mitral regurgitation (34.8% vs 11. 8%, P <.01). Women with echocardiographic MVP had higher ethnicity-adjusted hostility scores (19.9 vs 16.1, P <.05) than women with no MVP. Among 111 (2.7%) of 4136 participants reporting prior physician diagnosis of MVP, only 5 (0.45%) of 111 had definite echocardiographic MVP. CONCLUSIONS: These data document a low prevalence of definite echocardiographic MVP and suggest a constellation of anthropometric, physiologic, and psychobehavioral characteristics in young adults with echocardiographic MVP. Most definite echocardiographic MVP diagnoses were discordant with self-reported MVP status, and false-positive diagnoses of echocardiographic MVP were made more often in women and whites. PMID- 10467200 TI - Neurosurgical evacuation of intracranial hemorrhage after thrombolytic therapy for acute myocardial infarction: experience from the GUSTO-I trial. Global Utilization of Streptokinase and tissue-plasminogen activator (tPA) for Occluded Coronary Arteries. AB - BACKGROUND: Intracranial hemorrhage is an uncommon but very dangerous complication in patients receiving thrombolytic therapy for acute myocardial infarction. Neurosurgical evacuation is often an available treatment option. However, the association between neurosurgical evacuation and clinical outcomes in these patients has yet to be determined. METHODS: The GUSTO-I trial randomly assigned 41,021 patients with acute myocardial infarction to 1 of 4 thrombolytic strategies in 1081 hospitals in 15 countries. A total of 268 patients (0.65%) had an intracranial hemorrhage. We assessed differences in clinical characteristics, neuroimaging features, Glasgow coma scale scores, functional status (disabled: moderate or severe deficit; not disabled: no or minor deficit) and 30-day mortality rate between the 46 patients who underwent neurosurgical evacuation and the 222 patients who did not. RESULTS: Mortality rate at 30 days for all patients with intracranial hemorrhage was 60%; an additional 27% were disabled. Evacuation was associated with significantly higher 30-day survival (65% versus 35%, P <.001) and a trend toward improved functional status (nondisabling stroke: 20% versus 12%, P =.15). CONCLUSIONS: Although intracranial hemorrhage is uncommon after thrombolysis for acute myocardial infarction, 87% of patients die or have disabling stroke. Although not definitive, these data indicate that neurosurgical evacuation may be associated with improved clinical outcomes. Physicians treating such patients should consider early neurosurgical consultation and intervention in these patients. PMID- 10467201 TI - Ethnicity and long-term outcome after an acute coronary event. Multicenter Myocardial Ischemia Research Group. AB - BACKGROUND: The role of ethnicity on the long-term outcome after myocardial infarction is not fully understood. METHODS AND RESULTS: We analyzed the data from the Multicenter Study of Myocardial Ischemia in North America and Japan. We enrolled patients after acute myocardial infarction (MI) or unstable angina, with follow-up for 6 to 43 months, an average of 26 months. Among patients enrolled, there were 627 white, 158 black, and 109 Asian patients. Unadjusted cardiac events (cardiac death or nonfatal MI) were more frequent in black patients than in the other 2 ethnic groups (12% in blacks, 6. 4% in whites, 4.0% in Asians, P =.022). Although insulin-dependent diabetes, history of hypertension, and female sex were most frequent in black subjects, coronary angioplasty and thrombolysis at index event were done equally. After adjusting for several covariates, Cox analyses revealed that the black group was significantly associated with cardiac events (hazard ratio 6.5, P =.002). Subgroup analyses showed that the event rate among patients who had a higher educational level (6.1% in whites, 5.9% in blacks, and 7.0% in Asian, P =.94) or who were in a professional occupational class (5.7% in whites, 4.0% in blacks, and 4.8% in Asians, P = 1.0) was not different among the 3 ethnic groups. CONCLUSIONS: Blacks have an increased rate of cardiac events after MI, and a lower socioeconomic status may contribute to the adverse outcome in this ethnic group. PMID- 10467202 TI - Impact of availability of hospital-based invasive cardiac services on racial differences in the use of these services. AB - BACKGROUND: Reports indicate that black patients are less likely than white patients to receive invasive cardiac services after hospitalization for acute myocardial infarction (AMI). There is still uncertainty as to why racial differences exist and how they affect patient outcomes. This is the first study to focus on the availability of invasive cardiac services and racial differences in procedure use. Study objectives were to (1) document whether racial differences existed in the use of invasive cardiac procedures, (2) study whether these racial differences were related to availability of hospital-based invasive cardiac services at first admission for AMI, and (3) determine whether there were racial differences in long-term mortality rates. METHODS: A historical cohort study was conducted with discharge records from all acute care hospitals in New Jersey for 1993 linked to death certificate records for 1993 and 1994. There were 13,690 black and white New Jersey residents hospitalized with primary diagnosis of AMI. Use of cardiac catheterization within 90 days, revascularization within 90 days (percutaneous transluminal coronary angioplasty [PTCA] or coronary artery bypass graft surgery [CABG]), and death within 1 year after admission for AMI were the main outcome measures. Patterns for PTCA and CABG as separate outcomes were also studied. Hospital-based cardiac services available were described as no invasive cardiac services, catheterization only, or PTCA/CABG. To account for payer status and comorbidity differences, patients 65 years and older with Medicare coverage were analyzed separately from those younger than 65 years. RESULTS: Black patients aged 65 and older were generally less likely to receive catheterization and revascularization than white patients, regardless of facilities available at first admission. For patients younger than 65 years, the greatest differences between black and white patients in catheterization and PTCA/CABG use within 90 days after AMI occurred when no hospital-based invasive cardiac services were available. However, use of invasive cardiac procedures within 90 days after AMI was substantially increased if the first hospital offered catheterization only or PTCA/CABG services, among all patients, especially among blacks younger than age 65. No significant racial differences or interactions with available services were found in 1-year mortality rates. CONCLUSIONS: Availability of invasive cardiac services at first hospitalization for AMI was associated with increased procedure use for both races. However, use of invasive cardiac procedures was generally lower for black patients than for white patients, regardless of services available. Long-term mortality rates after hospitalization for AMI did not differ between blacks and whites. PMID- 10467203 TI - Thrombolysis with saruplase versus streptokinase in acute myocardial infarction: five-year results of the PRIMI trial. AB - BACKGROUND: Short-term safety and efficacy of thrombolysis with saruplase in acute myocardial infarction have been shown in several trials. To assess long term outcome of patients treated with saruplase or streptokinase for myocardial infarction, a 5-year follow-up of patients included in the Pro-Urokinase in Myocardial Infarction Trial was performed. METHODS AND RESULTS: Follow-up data are available from 8 centers on 255 (92.4%) of 276 included patients. The 5-year mortality rate was comparable with 20.8% of patients in the saruplase group and 16.9% in the streptokinase group (odds ratio 1.29, 95% confidence interval 0.69 to 2.42). In both groups, a considerable number of fatal cardiovascular events occurred more than 1 year after study inclusion. Rates of percutaneous transluminal coronary angioplasty and coronary artery bypass grafting were comparable in both groups. Reinfarction within 5 years occurred in 19.0% of patients in the saruplase group and tended to be less frequent at 10.8% after streptokinase treatment (odds ratio 1.94, 95% confidence interval 0.98 to 3.84). In both groups, the majority of reinfarctions took place more than 3 months after study inclusion. The 5-year stroke rate was 3.6% and 7.2% in the saruplase and streptokinase groups, respectively (odds ratio 0.49, 95% confidence interval 0.16 to 1.47). Subjective symptoms of heart failure and angina pectoris were comparable in both groups. CONCLUSIONS: Our data are consistent with a similar long-term outcome for patients treated with saruplase or streptokinase. Despite the low-risk profile of the patient cohort, there were considerable adverse event rates over a 5-year period. PMID- 10467204 TI - Continuous ST-segment monitoring associated with infarct size and left ventricular function in the GUSTO-I trial. AB - BACKGROUND: The aim of this study was to evaluate whether in patients with myocardial infarction, the intensity and duration of myocardial ischemia as measured by continuous ST monitoring are associated with infarct size and residual left ventricular function. METHODS AND RESULTS: The analyses included patients with myocardial infarction, receiving thrombolytic therapy, who were enrolled in the electrocardiographic substudy of GUSTO-I, monitored by a vector derived 12-lead electrocardiographic recording system, and in whom either infarct size (defined as cumulative release of alpha-hydroxybutyrate dehydrogenase activity per liter of plasma over a 72-hour period [Q(72)]) or left ventricular ejection fraction (LVEF) was determined. With the use of linear regression analysis, we investigated the association of various ST-trend characteristics with Q(72) (206 patients) and with LVEF (180 patients). A higher area under the ST trend since thrombolysis until 50% ST recovery and a higher area under recurrent ischemic episodes (ST reelevations) were significantly associated with a higher Q(72), whereas only a higher area under recurrent ischemic episodes was significantly associated with a lower LVEF. These associations remained after adjusting for other patient characteristics such as age, sex, infarct location, and time to treatment. CONCLUSIONS: These findings support the physiologic hypothesis that both the intensity and duration of myocardial ischemia (both reflected by the estimated areas under the ST-trend curve) determine myocardial damage and thus are associated with infarct size and ejection fraction in patients with acute myocardial infarction who receive thrombolytic therapy. PMID- 10467205 TI - Bedside markers of coronary artery patency and short-term prognosis of patients with acute myocardial infarction and thrombolysis. AB - BACKGROUND: In this study we have evaluated the prognostic power of noninvasive markers of coronary artery reperfusion in patients with acute myocardial infarction who were treated with intravenous streptokinase. METHODS: In 967 consecutive patients with acute myocardial infarction who were treated within 6 hours of symptoms, we analyzed the prognostic power of resolution of chest pain and ST-segment elevation >50% at 90 minutes, abrupt creatine kinase rise before 12 hours, and T-wave inversion in infarct-related electrocardiographic leads within the first 24 hours after thrombolysis. RESULTS: Global in-hospital mortality rate was 12.0%. Each reperfusion marker was associated with improved outcome. Multivariate logistic regression analysis showed that 3 of the 4 markers of coronary artery reperfusion were significantly and independently associated to low in-hospital mortality rate. The presence of early T-wave inversion was associated with the lowest in-hospital mortality rate (odds ratio 0.25, confidence interval 0. 10-0.56). When all markers of coronary artery reperfusion were included in the regression model, T-wave inversion (odds ratio 0.29, confidence interval 0.11-0.68) and abrupt creatine kinase rise (odds ratio 0.36, confidence interval 0.16-0.77) continued to be significantly associated with better outcome. CONCLUSION: A systemic analysis of noninvasive markers of coronary artery reperfusion can provide the clinician with an excellent tool to predict clinical outcomes when treating myocardial infarction. PMID- 10467206 TI - Sex and race differences in short-term prognosis after acute coronary heart disease events: the Atherosclerosis Risk In Communities (ARIC) study. AB - BACKGROUND: Case fatality after myocardial infarction (MI) among patients admitted to the hospital may differ between men and women and blacks and whites. Furthermore, a different pattern of sex and race differences in case fatality may occur when coronary deaths outside the hospital are included in the analysis. The ARIC study provides community-based data to examine 28-day case fatality rates after coronary heart disease (CHD) events. METHOD AND RESULTS: Surveillance of out-of-hospital CHD deaths and hospitalized MI was conducted in 4 U.S. communities from 1987 to 1993. Hospital discharges and death certificates were sampled, medical records abstracted, and interviews conducted with witnesses of out-of-hospital deaths. MI and out-of-hospital death classifications followed a standard algorithm. Linkage of hospitalized MIs to fatality within 28 days ensured complete ascertainment of case fatality rate. Comorbidities and complications during hospital stay were compared to assess possible explanatory factors for differences in case fatality. Overall, age-adjusted 28-day case fatality (MI plus CHD) was higher in black men compared with white men (odds ratio 1.78, 95% confidence interval 1.4-2.2) and in black women compared with white women (odds ratio 1.5, 95% confidence interval 1. 2-2.0). Although men had higher overall case fatality rates than did women, this difference was not statistically significant. After a hospitalized MI, 28-day case fatality rate was not statistically significantly different between men compared with women or blacks compared with whites. CONCLUSION: Race and sex differences in case fatality after hospitalized MI were not evident in these data, although when out of-hospital deaths were included, men and blacks were more likely than women and whites to die within 28 days of an acute cardiac event. A majority of deaths occurred before hospital admission, and additional study of possible reasons for these differences should be a priority. PMID- 10467207 TI - Relation between myocardial infarction, depression, hostility, and death. AB - OBJECTIVE: To examine the independent impact of major depression and hostility on mortality rate at 6 months and 12 months after discharge from the hospital in patients with a myocardial infarction. METHOD: Three hundred thirty-one patients were prospectively evaluated for depression with a modified version of the National Institute of Mental Health Diagnostic Interview Schedule for major depressive episode. The Cook Medley Hostility Scale data were analyzed by chi(2) procedures for nominal and categoric data, and Student t test was used for continuous data types. RESULTS: Depression was a significant predictor of death at 12 months (P =. 04) but not at 6 months (P =.08). Hostility was not found to be a predictor of death at 6 months or 12 months. CONCLUSIONS: Major depression in patients hospitalized after myocardial infarction is a significant univariable predictor of death at 12 months, although it was not a statistically significant predictor after adjusting for other variables. Hostility is not a predictor of death. Prospective studies are needed to determine the impact of aggressive treatment of depression on post-myocardial infarction survival. PMID- 10467208 TI - Increased expression of the CD11b/CD18 antigen on the surface of peripheral white blood cells in patients with ischemic heart disease: further evidence for smoldering inflammation in patients with atherosclerosis. AB - BACKGROUND: This study examined the availability of the CD11b/CD18 and CD62L antigens on the surface of peripheral blood leukocytes in patients with ischemic heart disease. METHODS AND RESULTS: The study population included 45 patients with angiographically documented ischemic heart disease admitted to our department of internal medicine and the cardiology department during 1 month (December 1997). Sixty-six healthy members of the hospital medical staff served as control subjects. Another 39 post-trauma patients who were admitted to the emergency room were also evaluated. Patients with ischemic heart disease had significantly (P <.002) higher concentrations of CD11b/CD18 antigen on the surface of their polymorphonuclear leukocytes and monocytes (mean fluorescence intensity of 203 +/- 81 and 261 +/- 75, respectively) compared with the control group (mean fluorescence intensity 158 +/- 68 and 211 +/- 74, respectively) and to the group of patients with acute stress (mean fluorescence intensity of 146 +/ 70 and 200 +/- 22, respectively). CONCLUSIONS: The presence of increased concentration of CD11b/CD18 suggests that circulating leukocytes are activated in patients with ischemic heart disease. This activation probably reflects the presence of an inflammatory response involving the atherosclerotic lesion and is not merely a result of acute stress. PMID- 10467209 TI - Diastolic potentials in verapamil-sensitive ventricular tachycardia: true potentials or bystanders of the reentry circuits? AB - BACKGROUND: Diastolic potentials (DP) are reported to be recorded in intracardiac electrograms during verapamil-sensitive ventricular tachycardia (VT) in which QRS complexes show complete right bundle branch block with a superior axis. The purpose of this study was to ascertain whether the DP recorded in the endocardial mapping during VT reflects the activation of the VT circuit. METHODS AND RESULTS: The study group consisted of 16 men and 2 women. The earliest activation site (EA site) was determined and the DP was recorded in the endocardial mapping during VT. We evaluated the response of the cycle length of VT, the interval between the ventricular activation and the DP (V-DP), and the interval between the DP and the ventricular activation (DP-V) to intravenous verapamil. Radiofrequency current was delivered to the EA site, the site where the DP was recorded, and the site where the DP and the Purkinje fiber potential of the left bundle branch (LB) were simultaneously recorded. In 15 patients, the DP was recorded in the wide posterior fascicle region of the LB. After verapamil, the cycle length of VT, the V-DP, and the DP-V were prolonged from 365 +/- 53 to 490 +/- 65, 315 +/- 30 to 368 +/- 30, and 50 +/- 27 to 123 +/- 36 ms, respectively, in 6 patients. The LB was recorded in all patients and the DP was recorded preceding the LB in 12 patients. VT was successfully ablated at the site where the DP and the LB were simultaneously recorded in all these patients. Ablation at the other sites failed. CONCLUSIONS: Radiofrequency ablation at the site where the DP was simultaneously recorded preceding the LB completely abolished the verapamil sensitive VT. The DP recorded with the LB simultaneously might reflect the slow conduction zone activity of the reentry circuit located within the Purkinje fiber network. PMID- 10467210 TI - Effect of exercise training on heart rate variability in healthy older adults. AB - OBJECTIVE: To determine the effect of exercise training on cardiac autonomic modulation in normal older adults by using analysis of heart rate variability. SUBJECTS: The exercise group consisted of 7 men and 9 women aged 66 +/- 4 years. The comparison group consisted of 7 men and 9 women also aged 66 +/- 4 years. METHOD: Heart rate variability was determined from 24-hour Holter recordings before and after 12 months of supervised exercise, which consisted of 3 months of stretching and 9 months of 5 hours/week aerobic exercise at approximately 70% of maximal oxygen uptake. Heart rate variability was measured at baseline and 12 months later in the comparison group, who had not changed their usual activity level. RESULTS: In the exercise group maximal oxygen consumption increased from 1.8 +/- 0.5 L/min to 2.2 +/- 0.7 L/min (P <.05). The standard deviation of normal interbeat intervals increased from 126 +/- 21 ms to 142 +/- 25 ms. Mean nighttime heart rate decreased from 67 +/- 6 beats/min to 63 +/- 5 beats/min. Increased fitness level had little effect on indexes of heart rate variability, which reflect parasympathetic or mixed sympathetic/parasympathetic modulation of heart rate. There was no change in heart rate or heart rate variability in the comparison group. CONCLUSIONS: Exercise training increases total heart rate variability in normal older adults. The most marked alterations are in nocturnal heart rate. Heart rate variability is stable over a 1-year period in older adults who do not alter their activity level. PMID- 10467211 TI - Long-term nitrate use may be deleterious in ischemic heart disease: A study using the databases from two large-scale postinfarction studies. Multicenter Myocardial Ischemia Research Group. AB - BACKGROUND: Secondary coronary prevention studies have generally focused on specific medications, often to the exclusion of commonly used therapies. To date, long-term nitrate use has not been investigated in large-scale clinical trials. METHODS AND RESULTS: We examined the relation between long-acting nitrates given during the chronic phase of the disease and the outcome. We analyzed data prospectively acquired in a large, observational study involving 1042 patients enrolled for the Multicenter Study of Myocardial Ischemia (MSMI) in North America, Israel, and Japan as well as 1779 patients enrolled for the Multicenter Diltiazem Post Infarction Trial (MDPIT). The Cox analyses with all the variables retained revealed that nitrates were associated with a significantly increased mortality risk (MSMI: hazard ratio 3.78, P =.011; MDPIT: hazard ratio 1.61, P =.019) in patients who had recovered from an acute coronary event. The analyses with the propensity score method on the MSMI and the MSMI databases also showed that the risk for cardiac death with use of nitrates was increased in most of the 5 subclasses according to the score. CONCLUSION: These analyses raise concern about the potential adverse effects of long-acting nitrate therapy in chronic coronary disease. PMID- 10467212 TI - Posttransplantation cytotoxic immunoglobulin G is associated with a high rate of acute allograft dysfunctions in heart transplant recipients. AB - BACKGROUND: The significance of anti-human leukocyte antigen immunoglobin G (IgG) detected in the posttransplantation course of heart graft recipients remains unclear. METHOD: Sera from 121 cardiac allograft recipients transplanted between January 1992 and December 1994 were screened for the presence of lymphocytotoxic antibodies in the first year after transplantation. Dithiothreitol was used to differentiate IgG from immunoglobulin M. RESULTS: Nineteen patients (15%) had cytotoxic IgG develop, mainly during the first month after transplantation. The percentage of women was higher in this group (42% vs 15.7%; P <.05). Donor to recipient mismatches for sex, blood typing, cytomegalovirus serology, and human leukocyte antigen typing were comparable between IgG producers and nonproducers. The frequency of acute allograft dysfunction during the first year after transplantation was significantly higher among patients producing IgG (42% vs 5.9; P <.001). Most of these acute allograft dysfunctions were independent of cellular rejection lesions but were associated with a thickening of the posterior wall and the interventricular septum during the acute episode. Finally, all the patients but one recovered. Recurrences were not uncommon and, at 1 year after transplantation, the dose of cyclosporine used in patients producing IgG was significantly greater, as was the left ventricular thickness. CONCLUSION: Posttransplantation cytotoxic IgG is not uncommon and appears to be associated with a high rate of acute allograft dysfunction. Development of these antibodies can be caused by a previous undetected immunization, as suggested by the higher percentage of women in the producer group. Correlation with histologic lesions of humoral rejection are discussed. PMID- 10467214 TI - Historic perspectives on the relative contributions of diastolic and systolic blood pressure elevation to cardiovascular risk profile. AB - The aim of this review was to examine the relative contributions of systolic and diastolic blood pressures to the risk of cardiovascular disease on the basis of epidemiologic evidence from the Framingham Heart Study and the change in attitudes toward systolic blood pressure that occurred during the course of the study. Historic texts were evaluated in comparison with data from the Framingham Heart Study, a prospective longitudinal analysis of the relation between blood pressure and occurrence of subsequent cardiovascular morbidity and mortality rates in a fixed cohort. Historically, systolic hypertension has been considered an innocent accompaniment to arterial stiffening, occurring as a compensatory phenomenon in the elderly. Epidemiologic data show that the development of hypertension is neither inevitable nor beneficial. The data also provide evidence that systolic pressure is more important than diastolic pressure as a determinant of cardiovascular sequelae. Mild or moderate elevations of systolic blood pressure, even when unaccompanied by diastolic pressure elevations, are associated with an increased risk of cardiovascular disease. Risk is increased further by the added presence of related metabolic disturbances such as dyslipidemia, glucose intolerance, insulin resistance, cardiac hypertrophy, and obesity. Over-reliance on diastolic blood pressure in assessing the risk of hypertension can be misleading. Systolic pressure constitutes a powerful predictor of cardiovascular disease and a valuable tool when incorporated within multivariate risk formulas for estimating the conditional probability of coronary and stroke events. PMID- 10467215 TI - Elevated systolic blood pressure and risk of cardiovascular and renal disease: overview of evidence from observational epidemiologic studies and randomized controlled trials. AB - The effect of elevated blood pressure on risk of cardiovascular and renal disease has been documented in both observational epidemiologic studies and clinical trials. However, these studies have traditionally concentrated on diastolic blood pressure to characterize the risk associated with hypertension. We reviewed evidence from prospective studies and randomized controlled trials to quantify the risk associated with systolic blood pressure. Prospective studies and randomized controlled clinical trials that were published in English-language journals were retrieved using MEDLINE, bibliographies, and the authors' reference files. All retrieved publications were reviewed and information on sample size, duration, study design, antihypertensive medication, participant characteristics, and outcomes was abstracted for randomized controlled trials that reported systolic blood pressure reduction during intervention. Several prospective studies indicate that the association between systolic blood pressure and risk of coronary heart disease, stroke, and end-stage renal disease is continuous, graded, and independent. Furthermore, they suggest that the association of systolic blood pressure with these outcomes is stronger than that of diastolic blood pressure. Pooling of the data available from randomized controlled trials indicates that an average reduction of 12 to 13 mm Hg in systolic blood pressure over 4 years of follow-up is associated with a 21% reduction in coronary heart disease, 37% reduction in stroke, 25% reduction in total cardiovascular mortality, and 13% reduction in all-cause mortality rates. These data indicate that systolic blood pressure is an independent and strong predictor for risk of cardiovascular and renal disease. PMID- 10467216 TI - Influence of arterial pulse and reflected waves on blood pressure and cardiac function. AB - An integrated view of the role of arterial blood pressure in cardiovascular physiology should consider both the steady (mean blood pressure) and pulsatile (systolic, diastolic, and pulse pressures) components. This brief overview describes the important factors influencing these components, with emphasis on the consequences of arterial stiffening. In addition to their conduit function, arteries also perform a cushioning function that transforms the pulsatile flow generated by contraction of the left ventricle into steady flow at the periphery. Arterial compliance is a principal determinant of arterial blood pressure and is both pressure dependent and affected by vascular biomechanics. Other important factors that affect the steady and pulsatile components of blood pressure include ventricle performance, peripheral resistance, pulse wave velocity, and the timing of pulse wave reflections. Ageing and hypertension are important factors that contribute to reductions in arterial compliance. Important functional effects of this are that both the amplitude of the arterial pulse wave and pulse wave velocity increase, causing an early return of reflected waves from the periphery to the aorta. This may boost aortic and left ventricular pressures during systole at the expense of diastolic pressure, which is reduced. Studies have shown that stiffening of arteries and the associated increase in systolic and pulse pressures are important cardiovascular risk factors. Patients with an increased risk of cardiovascular events associated with such changes should be identified and receive appropriate therapeutic interventions. PMID- 10467217 TI - Risk reduction associated with lowering systolic blood pressure: review of clinical trial data. AB - Hypertension is very common in adults >/=60 years of age. Isolated systolic hypertension (ISH) in particular is a good predictor of events associated with elevated blood pressure such as stroke, coronary heart disease, and congestive heart failure. Two large studies, the Systolic Hypertension in the Elderly Program (SHEP) and the Systolic Hypertension in Europe study (Syst-Eur), have demonstrated that antihypertensive drug therapy for elderly patients with ISH reduces the risk of stroke and other major cardiovascular events. SHEP demonstrated that antihypertensive drug treatment with a diuretic-based regimen in patients >/=60 years of age with ISH reduced the incidence of total stroke by 36%. SHEP also demonstrated a 32% reduction in the incidence of cardiovascular events for patients receiving active treatment. The 5-year average systolic blood pressure (SBP) was 155 mm Hg in the placebo group (n = 2371) compared with 143 mm Hg for the active treatment group (n = 2365). Results from the recently completed Syst-Eur study also support the benefits of antihypertensive treatment in patients >/=60 years of age with ISH. Active treatment in Syst-Eur consisted of the calcium channel blocker nitrendipine, with the addition of enalapril and hydrochlorothiazide as needed to reduce SBP to <150 mm Hg. In the active treatment group, total stroke decreased by 42%, and all cardiovascular events decreased by 31%. At 2 years, sitting SBP had decreased by 13 mm Hg in the placebo group (n = 2297) compared with 23 mm Hg in the active treatment group (n = 2398). PMID- 10467218 TI - Current clinical practice in hypertension: the EISBERG (Evaluation and Interventions for Systolic Blood pressure Elevation-Regional and Global) project. AB - Hypertension is one of the major treatable factors contributing to the burden of disease worldwide. However, despite national programs to encourage detection and treatment, there are still shortcomings in hypertension management. A large proportion of these can be attributed to socioeconomic factors, professional shortcomings, and patient noncompliance with management regimens. The Evaluation and Interventions for Systolic Blood pressure Elevation-Regional and Global (EISBERG) project was conceived to examine the reasons for suboptimal management of hypertension in more detail. Emphasis was placed on control of elevated systolic blood pressure because this is often neglected despite evidence that it is more important than diastolic pressure in predicting cardiovascular risk. The goals of the project include analysis of the relative importance of systolic and diastolic blood pressure as cardiovascular risk factors, identification of any necessary changes in practice, and the development and implementation of programs to promote appropriate changes in practice or attitudes. The three components of the initiative are a formal collection of epidemiologic evidence to examine the relation between systolic blood pressure and outcome; a cross-sectional, quantitative database (CardioMonitor) providing information on the treatment of patients with cardiovascular diseases including hypertension in seven countries, and qualitative research into hypertension management to assess attitudes, awareness, and knowledge among primary care physicians, patients and their carers. The study showed more effort was directed toward control of diastolic than systolic blood pressure. Adequate control of systolic blood pressure was seldom achieved. Blood pressure targets tended to be raised in elderly patients in conflict with recommendations and evidence that there is greater benefit in treating hypertension more aggressively in this population. Despite awareness of guidelines for treating hypertension, doctors were unsuccessful in practice. PMID- 10467219 TI - Safety and efficacy of eprosartan, a new angiotensin II receptor blocker. AB - Angiotensin-converting enzyme inhibitors have been used extensively in the management of hypertension and related cardiovascular conditions. However, this treatment approach is limited by lack of specificity and continued production of angiotensin II by other routes. Antagonism of the angiotensin II receptor offers the possibility of improved control of hypertension by providing a more complete blockade of the renin-angiotensin system than angiotensin-converting enzyme inhibition without bradykinin-related effects. Eprosartan is the only nonbiphenyl, nontetrazole competitive angiotensin II receptor antagonist clinically available and is highly selective for the AT1 receptor subtype. In clinical trials, eprosartan has been shown to lower blood pressure effectively in a once-daily regimen in hypertensive patients. In the recommended dose range of 600 to 1200 mg once daily, eprosartan is effective in patients with all grades of hypertension irrespective of age, sex, or race. Furthermore, the tolerability profile of eprosartan is comparable to that of placebo, and there are no known clinically relevant drug-drug interactions. A number of large-scale clinical studies are currently underway or planned to determine which of the increasing number of AT1 receptor antagonists may reduce cardiovascular morbidity and mortality rates in different groups of hypertensive patients. Meanwhile, current evidence suggests that the AT1 receptor antagonists represent a significant new approach to cardiovascular therapy and merit a fuller assessment in hypertension and other cardiovascular diseases. PMID- 10467220 TI - Pharmacology of eprosartan, an angiotensin II receptor antagonist: exploring hypotheses from clinical data. AB - Selective blockade of the angiotensin II AT1 receptor represents a novel mechanism for interrupting the renin-angiotensin system without altering the potential benefits of AT2 receptor stimulation. This selective inhibition produces none of the disadvantages associated with reduced bradykinin metabolism and angiotensin II generated by non-angiotensin-converting enzyme pathways. Eprosartan is a potent (1.4 nmol/L) AT1 receptor antagonist that competitively blocks angiotensin II-induced vascular contraction. In various animal models of disease, including hypertension and stroke, eprosartan is effective in reducing disease progression. Eprosartan also has sympathoinhibitory activity, as demonstrated by an inhibition of the pressor responses induced by activation of sympathetic outflow through spinal cord stimulation in pithed rats. In contrast, some of the other angiotensin II receptor antagonists, such as losartan, at equivalent angiotensin II blocking doses, have no effect on sympathetic nervous system activity. Because eprosartan can inhibit both the direct effects of angiotensin II as well as the indirect effects that are mediated by enhanced sympathetic neurotransmission, this may represent an important advance in the treatment of elevated systolic blood pressure. PMID- 10467221 TI - Integrated approaches to management of hypertension: promoting treatment acceptance. AB - Overwhelming trial evidence indicates that the treatment of hypertension is beneficial, but in practice, less than 50% of treated hypertensive subjects have blood pressure well controlled. The success of treatment relies on acceptance by the patient. Treatment acceptance may be affected by the efficacy and tolerability of drug therapy, its effects on quality of life, and other important but less well-recognized influences such as the expectations and preconceived ideas of the physician and the patient. This report briefly reviews the factors affecting patient concordance with antihypertensive treatment and the role these factors play in the development of an integrated treatment plan. Nonconcordance with drug therapy is common: Only one third of patients always take treatment, one third take it sometimes, and one third never take their prescribed medication. With poor concordance, control of blood pressure and the consequent benefits are less likely to be realized. The factors that influence concordance are ill understood. Although drug side effects and convenience of dosing regimens are contributors, the attitudes of patients, physicians, and their interactions are likely to be of considerable importance. Concordance may be improved by involving the patient in the treatment plan, setting explicit targets, following a clear treatment plan, motivating the patient to comply with treatment, paying attention to the concerns and particular needs of the individual patient, and by ensuring frequent contacts between patients and health care professions. Successful integrated approaches to the management of hypertension must address all the factors that affect treatment acceptance. PMID- 10467222 TI - Assimilating new therapeutic interventions into clinical practice: how does hypertension compare with other therapeutic areas? AB - Medical research has helped to clarify the benefits of some therapies for improving the treatment or outcome associated with cardiovascular disease. However, the adoption of these approaches into routine clinical practice is, in many cases, inadequate. Consequently, there are many missed opportunities to reduce the burden of morbidity and mortality from cardiovascular disease. This review summarizes the factors that may prevent modified behavior in medical practice and the effectiveness of interventions that influence change. There are many barriers that may prevent or slow the adoption of new therapeutic advances into routine clinical practice. As a result, the use of well-proven, efficacious therapy can be suboptimal. Because of this underuse, the realized benefits of treatment are below the potential benefits. Adoption of new therapies is highly dependent on the use of interventions to promote clinical change. However, the effectiveness of different types of interventions varies greatly. Nevertheless, there is a wide range of strategies available that can be used to induce real changes in practice performance and potentially improve patient outcomes. It is essential that future intervention strategies focus on improving adoption of new therapies into clinical practice. The physician must be encouraged to prescribe proven treatments to those patients who stand to benefit most. In addition, better systems of care should be developed that improve the identification of patients as suitable candidates for proven treatments and sustain their long-term commitment to therapy. PMID- 10467223 TI - Growth hormone, IGF-I and cancer. Less intervention to avoid cancer? More intervention to prevent cancer? AB - The GH/IGF-I axis has a clearly established role in somatic growth regulation and there is much evidence suggesting that it can play a contributing role in neoplastic tissue growth; a number of recent epidemiological reports indicate that it may also be an important determinant of cancer incidence. Whilst there have been previous reports of changes to the axis in patients with established cancers, these new studies are distinct in being prospective and the inferences that can be made from this are outlined in this review. The recent studies are considered within the context of other indirect epidemiological evidence, and together indicate that the GH/IGF-I axis may establish the level of predisposition to a number of common cancers and indeed that such risk may be programmed from early life. There is considerable evidence for a number of possible mechanisms, both direct and indirect, which could account for the associations between GH/IGF-I levels and cancer incidence; these mechanisms are briefly summarised. The implications of the new findings are then discussed in relation to the increasing clinical usage of chronic GH administration and the need for further studies to establish any consequent increase in cancer risk. Finally the opportunities for further work to optimise cancer risk assessment and risk reduction strategies are highlighted. PMID- 10467224 TI - Peroxisome proliferator-activated receptor gamma is induced during differentiation of colon epithelium cells. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma), a fatty acid activated nuclear receptor, is implicated in adipocyte differentiation and insulin sensitisation. In view of the association of dietary fat intake and bowel disease, the expression of PPARgamma in rodent and human intestine was studied. Expression of PPARgamma mRNA was examined by Northern blot hybridisation, RNase protection, and/or competitive RT-PCR assays, whereas PPARgamma protein levels were evaluated by immunoblotting and immunohistochemistry. PPARgamma mRNA and protein were abundantly expressed in colon relative to the small intestine both in rodents and in man. Interestingly, expression of PPARgamma was primarily localised in the more differentiated epithelial cells in the colon. The level of expression of PPARgamma in colon was similar to the levels seen in adipose tissue. Expression of PPARgamma increased from proximal to distal segments of the colon in man. In Caco-2 and HT-29 human adenocarcinoma cells, PPARgamma expression increased upon differentiation, consistent with PPARgamma being associated with a differentiated epithelial phenotype. High-level expression of PPARgamma was observed in the colon, but not in the small intestine, suggesting a potential role of this nuclear receptor in the colon. PMID- 10467225 TI - Androgen receptor expression of proliferating basal and luminal cells in adult murine ventral prostate. AB - Maintenance of the size and differentiated function of the adult prostate is dependent on testicular androgens. In this study, simultaneous androgen receptor (AR) immunohistochemistry and [(3)H]thymidine labelling was used to characterise the proliferating epithelial cells of the murine ventral prostate. Proliferation in the adult prostate was more prevalent in the basal cell population with 1.8&percent; AR-negative cells labelled with [(3)H]thymidine as compared with 0.7% AR-expressing luminal cells. Three weeks following castration of mice, the atrophied prostate contained rudimentary glands composed of both luminal and basal cells with the proportion of AR-expressing basal cells reduced from 50 to 25%. Administration of testosterone enanthate to castrated mice induced a recapitulation of the prostate gland that was preceded by up-regulation of AR expression in basal cells to normal adult levels (50% AR-positive cells) by 12 h following testosterone injection. Proliferation of AR-positive luminal cells peaked at 48 h (22.8%) while proliferation of AR-negative basal cells peaked at 96 h (6.1%) following testosterone administration. These results suggest that distinct populations of luminal and basal cells are resistant to castration induced involution of the prostate but remain responsive to direct or indirect testosterone effects and recapitulate the gland following administration of testosterone. PMID- 10467226 TI - Secretion of tissue inhibitors of matrix metalloproteinases by human fetal membranes, decidua and placenta at parturition. AB - At parturition, breakdown of extracellular matrix in the fetal membranes may play a part in the rupture of the membranes and in the aetiology of premature rupture, in addition to having a regulatory role in the cell-cell interactions and signalling at the feto-maternal interface to stimulate myometrial contractility. The matrix metalloproteinases (MMPs) are important enzymes for the breakdown of extracellular matrix and their activity is regulated by a family of endogenous inhibitors, the tissue inhibitors of matrix metalloproteinases (TIMPs). At parturition, alteration in the balance between MMPs and TIMPs may mediate this extracellular matrix breakdown during rupture of fetal membranes. The aims of this study were to determine if the intrauterine secretion of TIMPs changes at labour, and to characterise their cellular sources. A broad range of TIMP activities (27-30 kDa, 24 kDa and 21 kDa) were detected by reverse zymography in term amniotic fluid. There was a significant (P<0.05) decrease in the amount of TIMPs in amniotic fluid and their release with the onset of labour. The TIMPs were characterised by immunoblot as TIMPs-1, -2, -3 and -4. High levels of TIMPs were secreted by explants of chorio-decidua, decidua parietalis and placenta, with less being released by amnion. Immunolocalisation studies revealed a specific distribution pattern for each of the TIMP isoforms. Trophoblast cells of chorion laeve, decidua parietalis and placental syncytiotrophoblast demonstrated specific immunoreactivity for all four isoforms. TIMPs were also found bound to selective regions of extracellular matrix. The decrease in TIMPs during labour may permit increased breakdown of extracellular matrix in the fetal membranes and decidua at parturition, thus altering cell signalling at the feto-maternal interface and facilitating membrane rupture. PMID- 10467227 TI - Transport of IGF-I across epithelial cell monolayers. AB - Epithelial cells line the lumens of organs including the gastrointestinal tract, kidney tubules and respiratory airways, where they regulate the transport of electrolytes and the movement of macromolecules. The current study aimed to investigate the transport of IGF-I across epithelial cell barriers. Epithelial cell lines derived from gut (IEC-6), kidney (MDBK) and lung (Mv1Lu) were shown to possess high-affinity, functional receptors for IGF-I and formed tight junctions in monolayer culture. To investigate the transport of IGF-I, the three cell lines were grown on microporous filters in a bi-chamber system. In comparison with filters without cells, IEC-6 and Mv1Lu epithelial cell monolayers restricted the passage of (125)I-IGF-I and [(3)H]inulin, whereas the MDBK cells virtually occluded any passage of these molecules. Transport of (125)I-IGF-I across the epithelial cell monolayers was significantly less than that of [(3)H]inulin, suggesting that the binding of (125)I-IGF-I to high-affinity IGF receptors or IGF binding proteins retarded its transport. Moreover, (125)I-IGF-I transport was not inhibited by the presence of excess unlabelled IGF-I. Our findings provide evidence for the restricted diffusion of intact, free IGF-I across gut, kidney and lung epithelial cell monolayers via a paracellular or low-affinity transcellular pathway. PMID- 10467228 TI - Effects of dexamethasone on proliferation, activity, and cytokine secretion of normal human bone marrow stromal cells: possible mechanisms of glucocorticoid induced bone loss. AB - It is well documented that glucocorticoid excess causes bone loss, but the mechanisms of these effects remain poorly defined. To understand further the mechanisms of glucocorticoid-induced osteoporosis, we investigated the effects of glucocorticoids on bone formation and bone resorption by examining the proliferation, functional activities, and cytokine secretion of cultured human bone marrow stromal cells (hBMSC). Treatment with dexamethasone for 24 h at the concentration of 10(-8) M significantly suppressed [(3)H]thymidine incorporation and further inhibition was observed with longer treatment (8 days) or higher concentration (10(-7) M). Alkaline phosphatase activity of hBMSC was markedly stimulated with addition of dexamethasone (10(-8) M), to 191 +/- 22% (after 4 days) and 317 +/- 46% (after 7 days) of control. Dexamethasone (10(-8) M) treatment for 48 h decreased the incorporation of [(3)H]proline into collagenase digestible protein (CDP; 43.7+/-7.9% of control) and non-collagen protein (65.2+/ 8.4% of control), with a greater effect on CDP. Northern blot analysis indicated that alpha1(I)-collagen mRNA level was decreased by dexamethasone to 27.6 +/- 9.0% of the control value after 1 day of exposure, and to 55.2 +/- 6.2% after 7 days. Dexamethasone markedly suppressed basal production of interleukin (IL)-6 and IL-11 and that stimulated by parathyroid hormone (PTH), IL-1alpha, or tumour necrosis factor-alpha in a dose-dependent manner. These results suggest that the glucocorticoid-induced bone loss is derived at least in part via inhibition of bone formation, which includes the suppression of osteoblast proliferation and collagen synthesis. As both basal and PTH-stimulated production of IL-6 and IL-11 are decreased by dexamethasone, the increased bone resorption observed in glucocorticoid-induced osteopenia does not appear to be mediated by IL-6 or IL 11. PMID- 10467229 TI - Glucocorticoid and mineralocorticoid regulation of angiotensin II type 1 receptor binding and inositol triphosphate formation in WB cells. AB - Mineralocorticoids, glucocorticoids, and angiotensin II (AngII) act cooperatively to maintain body fluid homeostasis. Mineralocorticoids, such as aldosterone and deoxycorticosterone-acetate (DOCA), function synergistically with AngII in the brain to increase salt appetite and blood pressure. In addition, glucocorticoids increase AngII-induced drinking and pressor responses and may also facilitate the actions of aldosterone on salt appetite. The AngII Type 1 (AT1) receptor mediates many of the physiological and behavioral actions of AngII. This receptor is coupled to the G-protein Gq, which mediates AngII-induced inositol triphosphate (IP3) formation. The WB cell line, a liver epithelial cell line that expresses the AT1 receptor, was used to examine the cellular basis of glucocorticoid and mineralocorticoid regulation of AT1 function. In this study corticosterone and dexamethasone treatments increased the number of AT1 receptors by activating the glucocorticoid receptor (GR). This increase in AT1 binding resulted in enhanced AngII-stimulated IP3 formation. However, only supraphysiological doses of aldosterone or DOCA increased AT1 binding, and this effect also was mediated by GR activation. Furthermore, despite evidence that mineralocorticoids and glucocorticoids function together to increase AngII-stimulated actions in vivo, aldosterone and dexamethasone did not act synergistically to affect AT1 binding, Gq expression, or IP3 formation. These results indicate that GR activation, and the subsequent increases in AT1 binding and in AngII-stimulated IP3 formation, may represent a cellular mechanism underlying the synergy between adrenal steroids and AngII. PMID- 10467230 TI - Inhibin B is a more sensitive marker of spermatogenetic damage than FSH in the irradiated non-human primate model. AB - This study evaluated the effect of bilateral testicular irradiation (2 Gy) on reproductive hormones, testicular volume (TV) and sperm parameters in six adult cynomolgus monkeys. Hormone levels (FSH, inhibin B and testosterone (T)) were determined to find the most valuable endocrine marker of irradiation-induced damage. All parameters were analysed at weekly intervals for 14 weeks. Histological evaluation of both testes was performed at week 14 after irradiation when one monkey was castrated and at week 27 when the remaining five monkeys were bilaterally biopsied. A decrease in body weight, TV (30% of the pre-treatment size) and sperm count was observed after irradiation. Severe oligozoospermia was achieved throughout the study but azoospermia was recorded only occasionally. Histological evaluation revealed a heterogeneous picture with patchy arrangement of seminiferous tubules containing advanced germ cell types. An increase (P<0.05) in FSH levels and, to a lesser degree also in T levels, occurred several weeks after irradiation. Inhibin B levels showed a sharp decline (P<0.001) as soon as 1 week after irradiation. FSH and inhibin B did not return to baseline levels during the observation period. A negative correlation was found between FSH and inhibin B values (r=-0.35, P<0.001). Inhibin B correlated positively with testis volume (r=0.73, P<0.001) and sperm counts (r=0.55, P<0.01). In conclusion, this study shows that inhibin B represents an early and more sensitive marker of testicular damage than FSH. Furthermore, the rapid fall of inhibin B after irradiation suggests that this hormone is a direct parameter of premeiotic germ cell proliferation. PMID- 10467231 TI - Expression of G-protein-coupled receptor kinases in pregnant term and non pregnant human myometrium. AB - There is evidence for hormonal receptor desensitisation in human myometrium, but little is known about the mechanisms involved in the loss of myometrial response to agonists such as beta(2)-adrenergic agonists, prostaglandin gamma and oxytocin. It is well known that the receptors for these hormones are coupled to G proteins. The first step of receptor desensitisation is the phosphorylation of activated receptors by a G-protein-coupled receptor kinase (GRK). GRKs are members of a multigene family and the various subtypes differ in their localisation, regulation and mode of action. We have used Western blotting and reverse transcription PCR to identify the GRKs present in human myometrium from pregnant and non-pregnant women as well as in cultured human myometrial cells. We have found that human myometrium expresses the GRK subtypes 2, 4gamma, 5 and 6. On the other hand, GRK3 and the isoforms GRK4alpha, beta and delta were not found in myometrial tissue. Our data indicate that GRK2 is only expressed in pregnant term myometrium and is not found in non-pregnant tissue. Moreover, GRK6 appears to be expressed at a much higher level in pregnant term tissue than in non pregnant myometrium. Our observations suggest that GRK2 and GRK6 may contribute to the regulation of uterine contractility at term. Further work is necessary to determine whether GRKs and receptor desensitisation play a role in disorders of uterine contractility. PMID- 10467232 TI - LH down-regulates gonadotropin-releasing hormone (GnRH) receptor, but not GnRH, mRNA levels in the rat testis. AB - The demonstration of an inhibitory effect of gonadotropin-releasing hormone (GnRH) agonists upon steroidogenesis in hypophysectomized rats and the presence of mRNA coding for GnRH and GnRH receptors (GnRH-R) in rat gonads suggests that GnRH can act locally in the gonads. To assess this hypothesis, we investigated the effects of GnRH analogs, gonadotropins and testosterone on the levels of both GnRH and GnRH-R mRNA in the rat testis. Using dot blot hybridization, we measured the mRNA levels 2 to 120 h after the administration of the GnRH agonist, triptorelin. We observed an acute reduction of both GnRH and GnRH-R mRNAs 24 h after the injection (about 38% of control). However, the kinetics for testis GnRH R mRNA were different from those previously found for pituitary GnRH-R mRNA under the same conditions. Initially, the concentrations of serum LH and FSH peaked, then declined, probably due to the desensitization of the gonadotrope cells. In contrast, the GnRH antagonist, antarelix, after 8 h induced a 2.5-fold increase in GnRH-R mRNA, but not in GnRH mRNA, while gonadotropins levels were reduced. Human recombinant FSH had no significant effect on either GnRH or GnRH-R mRNA levels. Inversely, GnRH-R mRNA levels markedly decreased by 21% of that of control 24 h after hCG injection. Finally, 24 h after testosterone injection, a significant increase in GnRH-R mRNA levels (2.3 fold vs control) was found, but a reduction in the concentration of serum LH, probably by negative feedback on the pituitary, was observed. In contrast, GnRH mRNA levels were not significantly altered following testosterone treatment. Since LH receptors, GnRH-R and testosterone synthesis are colocalized in Leydig cells, our data suggest that LH could inhibit the GnRH-R gene expression or decrease the GnRH-R mRNA stability in the testis. However, this does not exclude the possibility that GnRH analogs could also affect the GnRH-R mRNA levels via direct binding to testicular GnRH-R. In contrast, the regulation of GnRH mRNA levels appeared to be independent of gonadotropins. Taken together, our results suggest a regulation of GnRH and GnRH R mRNA specific for the testis. PMID- 10467233 TI - Role of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptors in the control of prolactin, growth hormone and gonadotropin secretion in prepubertal rats. AB - Excitatory amino acids, such as glutamate, constitute a major transmitter system in the control of hypothalamic-pituitary secretion. Different subtypes of glutamate receptors, such as NMDA (N-methyl-d-aspartic acid) and KA (kainate) receptors, are involved in the control of anterior pituitary secretion. Other receptor subtypes, such as AMPA (activated by alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid) and metabotropic receptors, have been identified, although their role in the control of neuroendocrine function remains largely unknown. Recent reports have demonstrated the involvement of AMPA receptors in the control of the steroid-induced luteinizing hormone (LH) surge in female and growth hormone (GH) secretion in male rats. The aim of this study was to assess the potential role of AMPA receptors in the control of GH, prolactin (PRL), LH and follicle-stimulating hormone (FSH) secretion in prepubertal 23-day old rats. To this end, prepubertal female rats were injected with AMPA (2.5 or 5 mg/kg i.p.) or the antagonist of AMPA receptors 1,2,3,4-tetrahydro-6-nitro-2, 3 dioxo-benzo (f) quinoxaline-7-sulfonamide (NBQX; 0.25 or 0.50 mg/kg i.p.). Serum LH and FSH concentrations and hypothalamic LH-releasing hormone (LHRH) content remained unchanged after AMPA or NBQX administration. In contrast, serum PRL levels significantly decreased 15, 30 and 60 min after i.p. administration of AMPA and increased 120 min after NBQX treatment, whereas serum GH levels increased after AMPA treatment and decreased after NBQX administration. Considering that AMPA has been shown to activate a subset of kainate receptors, its effects were compared with those elicited by 2.5 mg/kg KA in prepubertal female rats. At this age, however, KA was unable to reproduce the effects of AMPA on PRL and GH secretion, thus suggesting that the actions observed after AMPA administration were carried out specifically through AMPA receptors. In addition, as the effects of AMPA on LH secretion in adult females have been proved to be steroid-dependent, the effects of AMPA (2.5 mg/kg) and NBQX (0.5 mg/kg) were tested in prepubertal animals with different gonadal backgrounds, i.e. intact males, and intact and ovariectomized (OVX) females. The effects of AMPA in prepubertal females appeared to be modulated by ovarian secretion, as the inhibition of PRL secretion disappeared and LH secretion was partially suppressed by AMPA in OVX animals whereas the stimulatory effect on GH release was enhanced by ovariectomy. Furthermore, in male rats, AMPA administration significantly decreased PRL secretion and increased serum GH levels, the amplitude of the GH response being higher than in prepubertal females. To ascertain the pituitary component for the reported actions of AMPA, hemi-pituitaries of male rats were incubated in the presence of AMPA (10(-8)-10(-6) M). The results obtained showed no effect of AMPA on PRL, GH and gonadotropin secretion in vitro. Finally, we investigated the involvement of the dopaminergic (DA) system in the inhibitory action of AMPA on PRL secretion. Pre-treatment of prepubertal female rats with a dopamine receptor antagonist (domperidone: 1 mg/kg) resulted in the blockage of AMPA-mediated inhibition of PRL secretion, thus suggesting that this action is probably mediated by an increase in DA activity. In conclusion, we provide evidence for the physiological role of AMPA receptors in the control of PRL and GH secretion in prepubertal rats. In contrast, our data cast doubts on the involvement of AMPA receptors in the regulation of gonadotropin secretion at this age. The effects of AMPA reported herein were not mediated through activation of kainate receptors and were probably exerted at the hypothalamic or suprahypothalamic levels. In addition, we show that ovarian secretion actively modulates the effects of AMPA receptor activation on anterior pituitary secretion in prepubertal female rats. PMID- 10467234 TI - Insufficient adaptive capability of pancreatic endocrine function in dexamethasone-treated ageing rats. AB - This study was aimed at exploring the capability of the pancreatic endocrine adaptive mechanisms of ageing Sprague-Dawley rats to counteract the metabolic challenge induced by the prolonged administration of dexamethasone (DEX) (0.13 mg/kg per day for 13 days). DEX treatment induced peripheral insulin resistance in 3-, 18- and 26-month-old rats, as indicated by the significant and persistent rise of plasma insulin levels in each age group (plasma insulin in 3-, 18- and 26 month-old rats from basal values of 4.3+/-0.8, 4.7+/-0.5 and 5.6+/-1.0 ng/ml (means+/-s.e.m.) respectively, rose to 11.9+/-1.7, 29.1+/-5.5 and 27.9+/-2.7 ng/ml respectively, after 9 days of administration). However, plasma glucose concentrations remained unchanged during the treatment in young rats, whereas they increased up to frankly diabetic levels in most 18-month-old and in all 26 month-old animals after a few days of DEX administration. Plasma free fatty acid concentrations increased 2-fold in 3- and 26-month-old rats and 4-fold in 18 month-old rats and could possibly be involved in the glucocorticoid-induced enhancement in insulin resistance, although they showed no significant correlation with glycaemic values. Incubation of pancreatic islets obtained from treated rats showed that DEX administration increased the insulin responsiveness of islets from not only younger but also older donors. However, in the islets of ageing rats, which already showed an age-dependent impairment of the sensitivity to glucose and other secretagogues, this enhancing effect was clearly attenuated with respect to the younger counterpart. Furthermore, DEX treatment depressed significantly the priming effect of glucose in islets isolated from all the three age groups. In conclusion, our results show that ageing rats are unable to counteract effectively a prolonged hyperglycaemic challenge as such induced by DEX administration. This homeostatic defect can be ascribed to the age-dependent failure of the endocrine pancreas to provide enough insulin to overcome the aggravation of an antecedent state of increased peripheral insulin resistance. PMID- 10467235 TI - Altered arterial concentrations of placental hormones during maximal placental growth in a model of placental insufficiency. AB - Pregnant ewes were exposed chronically to thermoneutral (TN; 20+/-2 degrees C, 30% relative humidity; n=8) or hyperthermic (HT; 40+/-2 degrees C 12 h/day, 35+/ 2 degrees C 12 h/day, 30% relative humidity, n=6) environments between days 37 and 93 of pregnancy. Ewes were killed following 56 days of exposure to either environment (days in treatment (dit)), corresponding to 93+/-1 day post coitus (dpc). Maternal core body temperatures (CBT) in HT ewes were significantly elevated above the TN ewes (HT; 39.86+/-0.1 degrees C vs TN; 39.20+/-0.1 degrees C; P<0.001). Both groups of animals displayed circadian CBT, though HT ewes had elevated amplitudes (HT; 0.181+/-0.002 degrees C vs TN; 0.091+/-0.002 degrees C; P<0.001) and increased phase shift constants (HT; 2100 h vs TN; 1800 h; P<0.001). Ewes exposed to chronic heat stress had significantly reduced progesterone and ovine placental lactogen (oPL) concentrations from 72 and 62 dpc respectively (P<0.05), corresponding to approximately 30 dit. However, when compared with the TN ewes, HT cotyledonary tissue oPL mRNA and protein concentrations were not significantly different (P>0.1). Prolactin concentrations rose immediately upon entry into the HT environment, reaching concentrations approximately four times that of TN ewes, a level maintained throughout the study (HT; 216.31+/-32.82 vs TN; 54. 40+/-10.0; P<0.0001). Despite similar feed intakes and euglycemia in both groups of ewes, HT fetal body weights were significantly reduced when compared with TN fetuses (HT; 514.6+/-48.7 vs TN; 703. 4+/-44.8; P<0.05), while placental weights (HT; 363.6+/-63.3 vs TN; 571.2+/-95.9) were not significantly affected by 56 days of heat exposure. Furthermore, the relationship between body weight and fetal length, the ponderal index, was significantly reduced in HT fetuses (HT; 3.01+/-0.13 vs TN; 3.57+/-0.18; P<0.05). HT fetal liver weights were also significantly reduced (HT; 27.31+/-4.73 vs TN; 45.16+/-6.16; P<0.05) and as a result, the brain/liver weight ratio was increased. This study demonstrates that chronic heat exposure lowers circulating placental hormone concentrations. The observation that PL mRNA and protein contents are similar across the two treatments, suggests that reduced hormone concentrations are the result of impaired trophoblast cell development, specifically trophoblast migration. Furthermore, the impact of heat exposure during maximal placental growth is great enough to restrict early fetal development, even before the fetal maximal growth phase (100 dpc-term). These data highlight that intrauterine growth retardation (IUGR) may result primarily from placental trophoblast cell dysfunction, and secondarily from later reduced placental size. PMID- 10467236 TI - Culture of prostate epithelial cells of the rhesus monkey on extracellular matrix substrate: influence of steroids and insulin-like growth factors. AB - Rhesus monkey prostate epithelial cells from the cranial lobe were isolated and cultured in flasks coated either with collagen IV or laminin. The effects of stromal cell medium, androgens and growth factors on cell number, thymidine incorporation and secretory activity were assessed. The results indicate that dihydrotestosterone (DHT) and androstenedione have stimulatory influences on cell proliferation and secretion in coated flasks. DHT was more effective in increasing cell number but the induction of secretory activity was similar with both steroids. The combination of IGF-I and -II resulted in inducing better cell proliferation and secretory activity than the individual IGFs but, of the two IGFs, IGF-I was more effective than IGF-II. DHT with IGFs was more potent in inducing proliferation, differentiation and secretion than androstenedione. Even in the absence of steroids or growth factors, colony formation and confluence occurred in coated flasks but cell differentiation and secretion only to a limited extent. In conclusion, we were able to establish an in vitro primary culture of prostate epithelial cells from rhesus monkey using extracellular matrix proteins, steroids and growth factors as additional supplements. This culture system may be useful to study prostate cell physiology and to identify drugs that can inhibit cell proliferation. PMID- 10467237 TI - Secretion of inhibin A, inhibin B and inhibin pro-alphaC during the oestrous cycle of the golden hamster (Mesocricetus auratus). AB - Plasma concentrations of inhibin pro-alphaC, inhibin A and inhibin B were determined by enzyme-linked immunosorbent assay at 6 h intervals throughout the 4 day oestrous cycle of the golden hamster. Plasma concentrations of follicle stimulating hormone (FSH) and oestradiol-17beta were also measured by radioimmunoassay during the oestrous cycle. Plasma concentrations of inhibin A increased from the early morning of day 1 (day 1=day of ovulation) and reached plateau levels at 0500 h on day 2. An abrupt increase in plasma concentrations of inhibin A was found at 1700 h on day 4, when the preovulatory FSH surge was observed. An increase in plasma concentrations of inhibin B occurred on day 1 and reached plateau levels at 1700 h on day 1. The levels remained elevated until 0500 h on day 4 and declined gradually by 2300 h on day 4. Plasma concentrations of inhibin pro-alphaC gradually increased with some fluctuation from day 1 to 1700 h on day 4 and then declined. Significant negative relationships were noted between plasma FSH and both dimeric forms of inhibin from day 1 to day 3. Significant positive relationships were found between plasma oestradiol-17beta and inhibin A or inhibin pro-alphaC throughout the oestrous cycle. In contrast, no significant relationship was found between plasma oestradiol-17beta and inhibin B. These findings suggest that both dimeric forms of inhibin play a role in the regulation of FSH secretion during follicular development. These findings also suggest that inhibin pro-alphaC could be secreted primarily by large follicles, and early atretic follicles could also be responsible for inhibin pro alphaC secretion. On the other hand, the secretory pattern of dimeric inhibins might shift from inhibin B to inhibin A with follicular development. PMID- 10467238 TI - Impaired adipogenesis in insulin-like growth factor binding protein-1 transgenic mice. AB - Differentiation of precursor cells into mature fat cells is accompanied by enhanced expression of insulin-like growth factor (IGF)-I and is stimulated by multiple hormones including growth hormone, glucocorticoids, IGF-I and insulin. We used transgenic mice that overexpress insulin-like growth factor binding protein-1 to investigate the role of IGF-I in the accumulation of fat tissue. In response to a sucrose-enriched diet, transgenic mice gained significantly less body weight and the epididymal fat mass was significantly reduced compared with wild-type mice. The increase in adipocyte size was also significantly reduced in transgenic mice compared with wild-type mice. Fewer colonies were generated from adipose tissue from transgenic mice and the mitogenic response of these cells to IGF-I was significantly reduced compared with those from wild-type mice. Induction of glycerol-3-phosphate dehydrogenase, a measure of adipocyte differentiation, by IGF-I but not insulin, was reduced in preadipocytes from transgenic mice. These data indicate that IGF-I has a critical role in the proliferation of adipocyte precursors, the differentiation of preadipocytes and the development of obesity in response to calorie excess. PMID- 10467240 TI - A modern anatomist at the helm of FASEB. PMID- 10467239 TI - LbetaT2 gonadotroph cells secrete follicle stimulating hormone (FSH) in response to active A. AB - Secretion of luteinizing hormone in response to gonadotropin releasing hormone (GnRH) has been described in the recently developed LbetaT2 gonadotroph cell line. We evaluated the expression of follicle stimulating hormone (FSH)beta mRNA and secretion of FSH from LbetaT2 cells in response to GnRH and activin A. LbetaT2 cells were treated with activin A in doses from 0 to 50 ng/ml, with or without a daily 10 nM GnRH pulse, or with GnRH alone. FSH secretion was stimulated over 6-fold by concomitant GnRH and activin A in a dose-responsive fashion at 72 h of treatment. FSHbeta mRNA was detectable by ribonuclease protection assay only in cells treated with activin A with or without GnRH. The demonstration of FSHbeta gene expression in LbetaT2 cells further validates these cells as mature, differentiated gonadotrophs and as an important tool for the study of gonadotroph physiology. PMID- 10467241 TI - Systematic bias in an "unbiased" neuronal counting technique. PMID- 10467242 TI - Keynote Address: The American Association of Anatomists' Sixth Annual Minority Student Luncheon, Experimental Biology '99, Washington, DC. PMID- 10467243 TI - Teaching the youngest anatomists. PMID- 10467244 TI - Going digital: image preparation for biomedical publishing. AB - Authors are more often being held responsible for readying their own data figures for digital publication by scanning them at the proper resolution and preparing them for presentation in both print and on-line journals. In this manner, the visuals can be printed at the highest quality the publisher can provide and be ready for rapid electronic distribution on the Internet. Therefore, authors must become knowledgeable in the visual preparation process in order to generate electronic images that will be as true a representation of the original image as possible. Perfecting this procedure can be a learning experience and often requires some experimentation. When accomplished, the author will have more control of exactly how the images will look before they are published. In addition to the scan resolution, the type of digital scanner and software applications used are very important, and instruction manuals should be followed closely so as to understand the full potential of the digitizing equipment. Anat Rec (New Anat): 257:128-136, 1999. PMID- 10467245 TI - Multipotent and restricted precursors in the central nervous system. AB - Acquisition of cell type-specific properties in the nervous system is likely a process of sequential restriction in developmental potential. At least two classes of pluripotent stem cells, neuroepithelial (NEP) stem cells and EGF dependent neurosphere stem cells, have been identified in distinct spatial and temporal domains. Pluripotent stem cells likely generate central nervous system (CNS) and peripheral nervous system (PNS) derivatives via the generation of intermediate lineage-restricted precursors that differ from each other and from multipotent stem cells. Neuronal precursors termed neuronal-restricted precursors (NRPs), multiple classes of glial precursors termed glial-restricted precursors (GRPs), oligodendrocyte-type 2 astrocytes (O2As), astrocyte precursor cells (APCs), and PNS precursors termed neural crest stem cells (NCSCs) have been identified. Multipotent stem cells and restricted precursor cells can be isolated from embryonic stem (ES) cell cultures providing a non-fetal source of such cells. Analysis in multiple species illustrates similarities between rat, mouse, and human cell differentiation raising the possibility that similar factors and markers may be used to isolate precursor cells from human tissue or ES cells. Anat Rec (New Anat): 257:137-143, 1999. PMID- 10467246 TI - Forthcoming topics PMID- 10467247 TI - BDNF and NT-4 differentially modulate neurite outgrowth in developing retinal ganglion cells. AB - We show here that neurite outgrowth of ganglion cells (RGCs) was selectively enhanced following treatment with BDNF or NT-4 in short-term cultures of dissociated cells derived from the neuroretina of postnatal rats. NT-4 was more effective than BDNF. The effect of NT-3 was variable, whereas NGF and CNTF had no effects upon neurite elongation. The neuritogenic responses of RGCs to both BDNF and NT-4 were prevented by competition with soluble TrkB receptor, and abolished by K252a, a selective inhibitor of the tyrosine kinase activity of Trks. These results indicate that the differentiating effects of BDNF and NT-4 are mediated by TrkB receptors, naturally expressed by RGCs. Developing RGCs treated with these TrkB ligands displayed distinct, albeit partially overlapping, patterns of neurite morphology. BDNF supported predominantly polarized outgrowth, whereas NT 4 induced the appearance of intensely branched symmetrical arbors. The lack of RGCs showing combined morphologies (e.g., highly arborized unipolar cells) suggests distinct mechanisms underlying either elongation or branching, and implicates distinct responses of RGC subsets. We conclude that neurite growth in vitro is extensively promoted by neurotrophins in developing RGCs. Moreover, highly homologous neurotrophins such as BDNF and NT-4, presumably activating via TrkB receptors, selectively control the differentiation of distinct ganglion cell neuritic morphologies. PMID- 10467248 TI - Expression of the bHLH gene NSCL-1 suggests a role in regulating cerebellar granule cell growth and differentiation. AB - We report that the neuronal-specific basic helix-loop-helix (bHLH) gene NSCL-1 is expressed at multiple and distinct stages of cerebellar granule cell differentiation. During embryonic development, NSCI-1 expression is initially evenly distributed in the cerebellar primordium and then becomes restricted to the ventricular zone. At the early steps of granule cell development, NSCL-1 is not expressed in rhombic lip cells, but instead in migrating granule cell precursors. Its expression culminates during postnatal proliferation of the external germinal layer, and remains only transiently in the newly formed internal granular layer, and at a much lower level. Thus, NSCL-1 expression is linked to the onset of granule cell differentiation, but is not involved in the maintenance of the differentiated state. These findings suggest that NSCL-1 does not behave as a specification factor, but rather as a factor promoting expansion of progenitor external germinal layer (EGL) cells. Gel mobility shift assays show that NSCL-1 only binds DNA as a heterodimeric complex with the ME1a E-protein. We also provide the first evidence that NSCL-1 functions as a transcriptional activator when heterodimerized with the ME1a E-protein. Taken together, these results suggest that NSCL-1 participates in the regulatory network controlling gene expression during cerebellar granule cell differentiation. PMID- 10467249 TI - Rhes: A striatal-specific Ras homolog related to Dexras1. AB - We have characterized an apparently full-length cDNA corresponding to a rat mRNA, SE6C, previously identified by subtractive hybridization as being expressed predominantly in the striatal region of the brain. The SE6C mRNA encodes a 266 amino acid protein with significant similarity to members of the Ras-like GTP binding protein family; thus, we have chosen the name Rhes, for Ras homolog enriched in striatum. The human homolog was found in a genomic sequence from human chromosome 22q13.1 and shares 95% identity with rat Rhes. Among the family of small G-proteins, Rhes shares 62% identity with Dexras1, a mouse dexamethasone inducible Ras-like protein. Both Rhes and Dexras1 have substantially longer C termini than other members of the Ras-like small G-protein family. Divergence between the C-terminal sequences of Rhes and Dexras1 suggests that, although their functions are probably similar, they have unique properties. Bacterially expressed Rhes binds GTP, suggesting that the protein indeed has GTPase functionality. Although Rhes was not induced by dexamethasone, its full expression is dependent upon thyroid hormone availability. Its accumulation is postnatal, consistent with the dependence upon thyroid hormone. It is noteworthy that most striatum-"specific" mRNAs characterized to date encode components of signal transduction cascades. PMID- 10467250 TI - Pigment epithelium-derived factor protects cultured retinal neurons against hydrogen peroxide-induced cell death. AB - Pigment epithelium-derived factor (PEDF) is a neurotrophic protein synthesized and secreted by retinal pigment epithelial (RPE) cells in early embryogenesis and has been shown to be present in the extracellular matrix between the RPE cells and the neural retina. It induces neuronal differentiation and promotes survival of neurons of the central nervous system from degeneration caused by serum withdrawal or glutamate cytotoxicity. Because the role of PEDF in the retina is still unknown, we examined its ability to protect cultured retinal neurons against hydrogen peroxide (H(2)O(2))-induced cell death. Retinas of 0-2-day-old Sprague-Dawley rats were isolated and dissociated, and the neurons were maintained for 2 weeks in a synthetic serum-free medium. Immunocytochemical labeling showed that 50-60% of the cultured cells were rod photoreceptors. Treatment with H(2)O(2) induced significant death of retinal neurons in a dose- and time-dependent manner. Pretreatment with PEDF prior to insult greatly attenuated H(2)O(2)-induced cytotoxicity, and its effect was shown to be dose dependent. Cytotoxicity was determined by 3,(4,5-dimethylthiazol-2-yl)2, 5 diphenyl-tetrazolium bromide and lactate dehydrogenase assays, and apoptotic cell death was evaluated by the TdT-mediated digoxigenin-dUTP nick-end labeling assay. The present study also showed that H(2)O(2)-induced retinal neuron death was by apoptosis that could be inhibited by PEDF. Combination of PEDF with basic fibroblast growth factor, brain-derived neurotrophic factor, or ciliary neurotrophic factor improves the protection. These data strongly suggest that PEDF is a potential neuroprotective agent in the treatment of retinal degeneration. PMID- 10467251 TI - Neuronal activity and transcription of proinflammatory cytokines, IkappaBalpha, and iNOS in the mouse brain during acute endotoxemia and chronic infection with Trypanosoma brucei brucei. AB - Trypanosoma brucei brucei (Tbb) infection is a model of chronic immune response associated with severe neurological disorders believed to lead to coma and death. We hypothesized that exaggerated production of proinflammatory molecules within the cental nervous system (CNS) may be involved in the etiology of the disease, i.e., African Tripanosomiasis. The purpose of the present study was therefore to verify the effects of the parasite Tbb on the genetic expression of the immediate early gene c-fos (index of cellular activity), tumor necrosis factor alpha (TNF alpha), interleukin-6 (IL-6), inhibitory factor kappa B alpha (IkappaBalpha, index of the nuclear factor kappaB activity, the transcription factor of numerous proinflammatory molecules), and inducible nitric oxide synthase (iNOS) in the mouse brain. Adult male BALB/c mice received a single intraperitoneal injection of lipopolysaccharide (LPS, used as positive control for these markers that are induced in a transient manner by the endotoxin), Tbb, or vehicle solution and were sacrificed at multiple times (1 hr to 7 days) following the injection. Acute and chronic models induced a robust expression of c-fos in numerous regions of the brain, including the circumventricular organs (CVOs) and different nuclei involved in autonomic control. Although the effect of LPS was rapid and transient, Tbb pathogen stimulated c-fos only within 5 to 7 days. The genes encoding TNF-alpha and IL-6 cytokines were expressed in the CVOs and choroid plexus 1 and 3 hr after LPS injection, whereas no convincing hybridization signal was detected in the brains of Tbb-infected mice at any time. IL-6 and iNOS expressing cells were also found along large blood vessels of LPS-treated mice, while scattered small TNF-alpha-expressing cells were observed across the brain 12 and 24 hr after the endotoxin treatment. Tbb caused a low to moderate expression of iNOS and IkappaBalpha genes in perivascular cells, but this effect was apparent only several days following the parasite infection. Taken together, these data indicate that LPS and Tbb stimulate c-fos expression in similar nuclei involved in autonomic control, an event occurring within the first 3 hr after the LPS insult and only 5 days post-Tbb injection. The mRNAs encoding proinflammatory cytokines were, however, not detected in Tbb-infected brains, which may be explained by the Tbb variant (MiTat 1.5) that caused high parasitaemias and mortality within 5 to 7 days. PMID- 10467252 TI - Differential regulation of cytoskeletal gene expression in hamster facial motoneurons: effects of axotomy and testosterone treatment. AB - We have previously demonstrated that systemic administration of testosterone increases the rate of axonal regeneration following facial nerve crush in adult male hamsters. In the present study, the molecular mechanisms by which androgens could enhance axonal regeneration were examined at a cellular level. Specifically, the following question was addressed using quantitative in situ hybridization with cDNA probes complementary to betaII, and alpha1 tubulin mRNAs: Does exogenous testosterone augment axotomy-induced changes in tubulin mRNA expression in hamster facial motoneurons (FMN)? Castrated adult male hamsters were subjected to right facial nerve severance, with the left side serving as internal control. One-half of the animals received testosterone replacement in the form of subcutaneously implanted silastic capsules containing crystalline testosterone propionate, and the other half were implanted with blank capsules immediately following the axotomy. Postoperative survival times from 2-14 days were examined. Axotomy alone resulted in a significant increase in the levels of both betaII and alpha1 tubulin mRNAs in facial motor neurons between 2-14 days after injury. Administration of testosterone selectively augmented the axotomy induced increases in betaII-tubulin, but not alpha1 tubulin, mRNA, levels at 7 and 14 days post axotomy. These results demonstrating an effect of testosterone in altering the neuronal cytoskeletal response to axotomy suggest that testosterone may enhance the regenerative properties of motor neurons via molecular mechanisms that involve selective alterations of the neuronal cytoskeleton. PMID- 10467253 TI - Peripheral nitric oxide contributes to both formalin- and NMDA-induced activation of nociceptors: An immunocytochemical study in rats. AB - Nociceptive c-fos expressions in the dorsal horn following intraplantar injection of two kinds of algogenic agents combined with different doses of nitric oxide (NO) synthase inhibitor (N(omega)-nitro-L-arginine methyl ester (L-NAME)) or of NO donor (L-arginine) were used to explore if NO was involved in the activation of peripheral nociceptors. The results showed that: 1) combined injections of L NAME with formalin into the plantar aspect of one hindpaw of normal rats elicited a dose-dependent suppression of c-fos expression as compared to that induced by formalin alone; 2) combined injections of L-arginine with formalin elicited considerable enhancement of c-fos expression when the dosages of L-arginine were less than 20 micromol, while it elicited marked suppression of c-fos expression when the dosages were in the range from 50 to 100 micromol; and 3) combined injection of L-NAME with NMDA, a selective agonist for NMDA receptors, into the hindpaw could also inhibit the NMDA-induced c-fos expression in the spinal dorsal horn. These results suggest that endogenously generated concentrations of nitric oxide may enhance the initiation of nociceptive inputs of peripheral nociceptors following local injection of formalin or NMDA. PMID- 10467254 TI - Dietary restriction and 2-deoxyglucose administration reduce focal ischemic brain damage and improve behavioral outcome: evidence for a preconditioning mechanism. AB - Stroke, an age-related disorder involving degeneration of neurons resulting from cerebral ischemia, is a major cause of disability and mortality. Although dietary restriction (DR) extends lifespan and reduces levels of cellular oxidative stress in several different organ systems including the brain, the impact of DR on ischemic brain injury is unknown. We report that maintenance of adult rats on a DR regimen resulted in reduced brain damage and improved behavioral outcome in a middle cerebral artery occlusion-reperfusion (MCAO-R) stroke model. Administration of 2-deoxyglucose (2-DG), a nonmetabolizable analogue of glucose, to rats fed ad libitum resulted in reduced ischemic brain damage and improved behavioral outcome following MCAO-R. 2-DG protected cultured hippocampal neurons against chemical hypoxia, demonstrating a direct protective action on neurons. DR and 2-DG administration resulted in an increase in the level of the stress protein heat-shock protein 70 (HSP-70) in striatal cells in vivo, and 2-DG treatment induced HSP-70 in cultured neurons suggesting involvement of a preconditioning stress response in the neuroprotective actions of DR and 2-DG. The neuroprotective effect of DR and 2-DG in this focal cerebral ischemia model suggests that outcome following stroke may be improved in individuals who follow a regimen of reduced food intake. PMID- 10467255 TI - AP1 transcriptional factor activation and its relation to apoptosis of hippocampal CA1 pyramidal neurons after transient ischemia in gerbils. AB - The cellular processes with a potential to lead to delayed death of neurons following transient (5 min) ischemia in gerbil hippocampus were evaluated. Neuronal apoptosis, visualized by the terminal transferase dUTP nick-end labelling (TUNEL) reaction, selectively appeared in the CA1 region of the pyramidal cell layer between the third and fourth days after the insult. Concomitantly, an enhanced immunoreactivity to anti-cJun/AP1 (N) antibody as a major component of activator protein 1 (AP1) transcriptional factor was observed in CA1 neurons. In contrast, in the early postischemic phase, the cJun/AP1 reaction was noticed in numerous neurons and glia-like cells of the CA2/CA3 region, hilus of the dentate gyrus, and region of mossy fiber terminals. In parallel, hippocampal protein binding to AP1, measured by the electrophoretic mobility shift assay (EMSA), showed biphasic enhancement at 3 and then 72-120 hours after ischemia. Supershifts, with antibodies against c-Fos and phospho-c Jun constituencies of the AP1 dimer, revealed an increased amount of phosphorylated c-Jun in the late postischemic phase. Collectively, these results suggest diversity of AP1 complex function, regulated by its dimer composition as well as time and place of expression during postischemic reperfusion. The early, survival-supporting AP1 response, located mainly in ischemia-resistant areas of CA2/3, is followed by the delayed phase, characteristic of massive neuronal apoptosis in CA1 with concomitant increase of phospho-c-Jun in AP1 dimer. PMID- 10467256 TI - Activation of ERK1 and ERK2 is required for manganese-induced neurite outgrowth in rat pheochromocytoma (PC12) cells. AB - Mn(2+) treatment has been shown to promote neurite outgrowth in rat pheochromocytoma (PC12) cells in a time- and dose-dependent manner. This process is mediated through the interactions of extracellular matrix (ECM) proteins and integrin receptors. Studies were performed to determine whether the phosphorylation of the MAP kinases, ERK1 and 2, is required for Mn(2+)-induced neurite outgrowth. A time- and dose-dependent increase in phosphorylation of both ERK1 and 2 was observed upon treatment of PC12 cells with Mn(2+). Phosphorylation of the ERKs occurred as early as 2 hr after initiating treatment, with a maximum increase occurring at approximately 24 hr. Inhibition of MEK with the specific inhibitor, PD98059, blocked the phosphorylation of ERK1 and 2 and increased Mn(2+) toxicity. When cells were grown in serum-free defined medium, Mn(2+) induced phosphorylation of ERK1 and ERK2 occurred in cells grown on surfaces treated with growth serum or fibronectin but not on surfaces treated with poly-L lysine. In addition, the pentapeptide GRGDS, which blocks RGD-mediated interactions, inhibited Mn(2+)-induced phosphorylation of ERK1 and 2. The Mn(2+) induced increase in phosphorylated ERK1 and 2 was not seen in a PC12 cell line that does not respond to Mn(2+). These data support the hypothesis that integrin mediated activation of the MAPK signal transduction pathway leading to the activation of ERK1 and 2 is required for Mn(2+)-induced PC12 differentiation and neurite outgrowth. PMID- 10467257 TI - Distinctions between microglial cells and peripheral macrophages with regard to adhesive activities and morphology. AB - Activated microglial cells and peripheral macrophages are hardly distinguishable from the viewpoints of morphology and function. There are various immunological markers common to both microglial cells and peripheral macrophages. In the present study, however, we found that microglial cells have distinct characters in terms of adhesion and morphology. By using a "rheoscope," that is an apparatus to rheologically measure the strength of cell adhesion to substrates, rat microglial cells were found to attach to polystyrene dishes much more weakly than alveolar and peritoneal macrophages. Interferon-gamma (IFNgamma) strengthened the adhesion of alveolar and peritoneal macrophages, whereas it weakened that of microglial cells. Morphological changes of microglial cells induced by IFNgamma were also different from those of peripheral macrophages. Furthermore, alveolar and peritoneal macrophages produced NO in response to IFNgamma, while microglial cells did not. When cultured on astrocyte-derived extracellular matrix (AsECM) in serum-free medium, only microglial cells extended multiple ramified processes. Conversely, alveolar and peritoneal macrophages on AsECM shrunk their ruffling membrane and rounded up. These distinctions between microglial cells and macrophages may reflect differences in cell lineages as well as environments in which individual cells reside. PMID- 10467258 TI - Effect of osmolality and anion channel inhibitors on myo-inositol efflux in cultured astrocytes. AB - Recent studies have shown that swelling-activated myo-inositol efflux from rat C6 glioma cells is mediated by a single transport mechanism and most likely by a volume-sensitive anion channel. In those studies, cells were acclimated in hypertonic medium and then swollen by returning the cells to isotonic medium. In the present study, myo-inositol efflux was determined in primary cultures of astrocytes by first incubating the cells in isotonic radiolabelled medium for 2 hr and then placing the cells in either unlabelled isotonic, hypertonic, or hypotonic medium and measuring release with time. Computer analyses of efflux data indicated a two-component system of myo-inositol efflux. The rate constants for the initial fast component for isotonic and hypotonic cells were 0.0398 +/- 0. 005 and 0.0631 +/- 0.0288 min(-1), respectively. The efflux rates of the slow component, while quite small, were severalfold greater with increasing hypotonic media as compared to the cells in isotonic medium. Several anion membrane transport inhibitors were tested to explore the swelling activated efflux mechanism of myo-inositol. Furosemide (0.5 mM), 1,9 dideoxyforskolin (0.1 mM), NPPB (0.1 mM), niflumic acid (0.5 mM), and SITS (0.5 mM) blocked the fast component of myo-inositol efflux by 17, 49, 55, 75, and 93%, respectively. Our results suggest that the fast component of myo-inositol efflux in primary cultures of astrocytes is mediated by anion transporters or channels and that myo inositol flux contributes to cell volume regulation in cultures of primary astrocytes. PMID- 10467259 TI - Rat alpha(2)-macroglobulin inhibits NGF-promoted neurite outgrowth, TrK phosphorylation, and gene expression of pheochromocytoma PC12 cells. AB - Rat alpha-1-macroglobulin (alpha(1)M) and alpha-2-macroglobulin (alpha(2)M) are murine homologs of human alpha(2)M, and rat alpha(2)M is generally known as an acute-phase protein. Monoamine-activated forms of human alpha(2)M have been shown to inhibit various neuronal functions, but the effect of rat alpha(1)M and acute phase alpha(2)M on neurons is largely unknown. In this report, rat serotonin activated alpha(2)M (5HT-alpha(2)M) has been demonstrated to inhibit nerve growth factor (NGF)-promoted neurite extension in pheochromocytoma PC12 cells, and we investigated its possible mechanism of action including its effect on NGF promoted signal transduction and gene expression in these cells. Especially in the absence of NGF, 5HT-alpha(2)M was found to bind to TrkA (the high-affinity receptor for NGF) much better than normal alpha(2)M (N-alpha(2)M). 5HT-alpha(2)M dose-dependently inhibited NGF-promoted autophosphorylation of TrkA, and decreased the expression of two immediate-early genes (NGFI-A and c-jun) and two delayed-response genes (SCG10 and transin) which are associated with neurite outgrowth in PC12 cells. The unmodified N-alpha(2)M, on the other hand, exhibited very little or no inhibitory effects on neurite extension, Trk phosphorylation, or expression of these genes. The results of this study taken together suggest that monoamine-activated acute-phase rat alpha(2)M appears to inhibit neurite outgrowth in PC12 cells possibly via its direct binding to TrkA and subsequent blocking of TrkA-mediated signal transduction and gene expression. PMID- 10467261 TI - In vivo extracellular electrophysiology of pallidal neurons in dystonic and nondystonic hamsters. AB - In the dt(sz) hamster, a model of idiopathic paroxysmal dystonia, recent findings indicated a decreased neuronal activity within the globus pallidus (GP) and an impaired gamma aminobutyric acid (GABA)ergic function when compared to nondystonic controls. Therefore, in the present study, extracellular single-unit recordings combined with systemical application of a subconvulsant prodystonic dose of pentylenetetrazole (PTZ) were used to compare the electrophysiological properties of GP neurons in anesthetized dt(sz) hamsters and nondystonic controls. The spontaneous discharge rate of GP neurons was not decreased but a trend towards a wide-ranged distribution was found in mutants compared to controls. Since the single-unit activity of striatal neurons was recently shown to be significantly increased in dt(sz) hamsters, the lack of significant changes in GP discharge rates was unpredicted. We suggest that this is due to antagonistic convergent striatal and subthalamic inputs and to lateral monosynaptic inhibition known for striatum and GP. While no significant changes of the discharge rate of GP neurons could be detected, the spike morphology was significantly altered in dt(sz) hamsters, suggesting subtle impaired information processing in the GP. The lack of marked changes in basal firing pattern may be related to the anesthesia. Administration of PTZ (25 mg/kg i.p.) at a subconvulsant dose, which aggravates dystonia in awake dt(sz) hamsters, seemed to induce more marked changes in spike morphology and firing pattern in mutants than in controls, although the discharge rate did not differ significantly between both animal groups in response to PTZ. In view of recent findings, we assume that GABAergic dysfunctions in dystonic hamsters are of regionally different extent. PMID- 10467260 TI - Overexpression of Akt (protein kinase B) confers protection against apoptosis and prevents formation of ceramide in response to pro-apoptotic stimuli. AB - An immortalized dorsal root ganglion cell line F-11 exhibits many properties of spinal cord neurons and undergoes apoptosis in response to growth factor withdrawal and the exogenous addition of inhibitors of phosphatidylinositol-3 kinase (PI3K). To elucidate the mechanism of apoptosis we generated F-11 clones which overexpressed either the p110 subunit of PI3K, a constitutively active form of protein kinase B/Akt (Myristoylated Akt), or a dominant-negative form (c-Akt). The first two constructs were protective against apoptosis induced by PI3K inhibitors such as wortmannin and LY294002. Caspase-3 (CPP32) levels peaked at 4 hr to 6 hr in response to pro-apoptotic drugs, and this increase was attenuated by 50% in F-11 with constitutively active Akt. The Akt protection was confirmed by DNA fragmentation studies. Both neo-transfected and the c-Akt dominant negative transfected F-11 cells showed increased ceramide formation (twofold) in response to staurosporine, wortmannin, or LY294002; whereas cells with a constitutively active Akt (Myr-Akt) showed no increase in ceramide when treated with staurosporine, wortmannin, or LY294002. Ceramide was a more potent activator of CPP32 and an inducer of apoptosis when added as the native form (hydroxy- or nonhydroxy-), rather than the more water-soluble C(2)-ceramide. Overexpression of PI3K (p110) and Akt protected cells against ceramide-induced apoptosis, suggesting that Ceramide action is upstream of Akt in these cells and suggesting that Akt might be a target for inhibition by ceramide. Both staurosporine and C(2)-ceramide activated the Jun kinase (JNK) cascade and C(2)-ceramide increased caspase-3 (CPP32) activity in cells expressing wild-type c-Jun, but not dominant negative (TAM-67) c-Jun. We suggest that this pathway is also involved in apoptosis, consistent with the idea that ceramide has multiple kinase and kinase modulating targets in the apoptotic pathway of neurons. J. Neurosci. Sci. 57:884 893, 1999. PMID- 10467262 TI - Role of intracellular Ca(2+) stores shaping normal activity in brain. AB - The role of intracellular Ca(2+) stores in the control of brain activity was investigated in microdialysis experiments by monitoring changes in the extracellular concentration of amino acids (AA) in the hippocampus of the rat after intracerebroventricular (icv) administration of the intracellular Ca(2+) release blocker, dantrolene in vivo, as well as in D-aspartate release and transmembrane Ca(2+) flux measurements in dantrolene-treated (50 microM) hippocampal homogenates containing resealed plasmalemma fragments and nerve endings in vitro. Microdialysis data demonstrate that icv injection of 0.6 mM dantrolene significantly decreases ( approximately 20%) the background (Glu) in the hippocampus. Both the (Glu; approximately 300%) and the inhibitory effect of dantrolene thereupon ( approximately 50%) was significantly increased when 0.5 mM of the Glu uptake inhibitor, L-trans-pyrrolidine-2,4-dicarboxylic acid, was dialysed into the hippocampus. NMDA and (S)-AMPA induced [(3)H]-D-aspartate release in hippocampal homogenates. Preincubation of these homogenates with 50 microM dantrolene was found to reduce the response to NMDA, but not to (S)-AMPA, in a NMDA-dependent manner. Increased rates of transmembrane influx and efflux of Ca(2+) in hippocampal homogenates with half-times of 4 ms and 200 ms, respectively, can be observed by the addition of 100 microM NMDA as recorded using a stopped-flow UV/fluorescence spectrometer in combination with the Ca(2+) indicator dye, bisfura-2. Both the Ca(2+) influx and efflux rates of the NMDA response were reduced (25-fold and >5-fold, respectively) in homogenates preloaded with 50 microM dantrolene. These results suggest a role for NMDA inducible intracellular Ca(2+) stores in the control of normal brain activity in vivo. PMID- 10467263 TI - Effect of dexamethasone on elevated cytokine mRNA levels in chemical-induced hippocampal injury. AB - An acute administration of the hippocampal toxicant trimethyltin (TMT) produced a specific pattern of neuronal necrosis in dentate granule cells with accompanying astrogliosis and initiation of a cytokine response within 24 hours. The purpose of this study was to examine the effects of the anti-inflammatory agent, dexamethasone (DEX), on the pattern of cytokine expression and neuronal degeneration occurring after an acute TMT injection. Dexamethasone (0.2 mg/kg or 10 mg/kg) was administered to 21-day-old male mice 1 hour prior to an injection of TMT hydroxide (2.5 mg/kg, i.p.). Mice receiving 0.2 mg/kg DEX received a second injection 6 hours after TMT. Twenty-four hours later, neuronal necrosis and astrogliosis were assessed and found to be similar in animals treated with TMT, either in the presence or absence of dexamethasone. Pretreatment with dexamethasone failed to prevent the neurodegeneration and astrogliosis. The TMT induced injury response was represented in elevations of mRNA levels for the injury-associated host response genes glial fibrillary acidic protein (GFAP), EB22/5.3, and intercellular adhesion molecule-1 (ICAM-1). The combination of DEX and TMT produced increased elevation in mRNA levels for EB22/5.3 and ICAM, while GFAP levels remained the same as with TMT alone. The injury response from TMT was accompanied by elevations in mRNA levels for the cytokines tumor necrosis factor (TNF) alpha, TNFbeta, and interleukin (IL)-1alpha. Treatment with dexamethasone prior to TMT resulted in significantly elevated levels of TNFalpha, TNFbeta, and IL-1alpha as compared to TMT alone. These data represent the inability of glucocorticoids to downregulate the injury response in rat hippocampus following a systemic injection of TMT and suggest a stimulation and "priming" of hippocampal cells by dexamethasone. PMID- 10467264 TI - In vitro neuroprotection against glutamate toxicity provided by novel non competitive N-methyl-D-aspartate antagonists. AB - Primary cultures of embryonic cortical cells were used to test the neuroprotective effect of two novel non-competitive N-methyl-D-aspartate antagonists against the neurotoxicity of glutamate. These two compounds, GK 11 and GK 222, are phencyclidine derivatives that contain asymetric carbon atoms. We have tested the neuroprotective effects of the racemates as well as those of the corresponding plus and minus enantiomers and have compared them with the reference drug dizocilpine (MK-801). We show here that the (-) enantiomers have a significantly better protective effect than the (+) enantiomers, but that the former are as efficient as the corresponding racemates. Qualitative analysis with different neuronal and glial markers does not reveal any difference in the population of protected cells among the different molecules tested. PMID- 10467265 TI - Therapeutic and "dose-dependent" effect of repetitive microelectroshock induced by transcranial magnetic stimulation in Parkinson's disease. AB - Transcranial magnetic stimulation (TMS) has been used in the diagnosis of neurological lesions and has been introduced into the therapy of central nervous diseases. Lately it has been claimed that TMS would be useful not only in the treatment of depression, but also in relieving symptoms of Parkinson's disease. In this study, we sought evidence of the effect of repetitive TMS on the symptoms of Parkinson's disease, the dose dependency between the applied elecromagnetic field and the Parkinsonian symptoms, and the maintenance of the improvement. Forty-nine patients with Parkinson's disease were divided into four groups, each given one stimulus, repeated 30 times, once or twice a day ( approximately 0.34Tesla (T), approximately 0.57T, approximately 0.80T). Patients were followed for 3 months and assessed using two different parkinsonian scales: the graded clinical rating scale and Unified Parkinson Disability Rating Scale (UPDRS), and with a short-term memory test (Ziehen-Ranschburg word pair test). No effect was seen in the group treated with approximately 0.34T?30 stimuli once a day. In all of the groups receiving TMS twice a day, the parkinsonian scores were significantly decreased compared with that of baselines after 1 month of treatment. The greatest improvement in the hypokinesia was detected in the group treated with approximately 0.57T?30 stimuli twice a day (baseline total UPDRS: 30.62 +/- 15.23; 1 month after treatment: 17.08 +/- 7.04, P < 0.01; 3 months after treatment: 16.08 +/- 7.06, P < 0.01). A dose-dependent difference was observed between the two groups after 3 months. The total UPDRS in Group II ( approximately 0.34T?30 stimuli twice a day) significantly differed from Group III ( approximately 0.57T?30 stimuli twice a day; 22.43 +/- 8.87, 16.08 +/- 7.06, P < 0.05). The long-lasting improvement effect with TMS would seem to suggest it as an appropriate tool in the therapy of Parkinson's disease. PMID- 10467266 TI - Regional heterogeneity of the astroglial immunoreactive phenotype: effect of lipopolysaccharide. AB - In order to find out whether the concept of regional heterogeneity in astrocytes also applies to the immunoreactive phenotype, we studied cultured primary rat astrocytes originating from five different brain regions (cortex, hippocampus, striatum, septum, and brainstem).We investigated this heterogeneity through the ability of lipopolysaccharide (LPS) to differentially induce several parameters that are known to characterize activated astroglia: major histocompatibility complex (MHC) class II and intercellular adhesion molecule (ICAM)-1 expression, nitric oxide (NO) production, synthesis of interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha). Under basal conditions, some of these parameters are already heterogeneic. The presence of LPS enhances these differences: expression of MHC class II increases after a 48-hour incubation with LPS, with brainstem and hippocampus astrocytes reaching the highest levels; NO production is induced by an LPS incubation, with the brainstem showing low NO production levels; septum and striatum instead show higher cytokine (TNF-alpha, IL-6) productions. The baseline expression of ICAM-1 also shows major regional differences, with the brainstem displaying the highest ICAM-1 expression. Our results demonstrate that the immunoreactive abilities of astrocytes show regional heterogeneities. This specialization may be implicated in the pathophysiological pathways of several neurodegenerative disorders. PMID- 10467267 TI - Axotomy-induced changes in pituitary adenylate cyclase activating polypeptide (PACAP) and PACAP receptor gene expression in the adult rat facial motor nucleus. AB - It has been demonstrated that pituitary adenylate cyclase activating polypeptide (PACAP) promotes the survival of neurons in culture and can inhibit neuronal cell death after experimental injury. Furthermore, peripheral axotomy results in increased PACAP gene expression in sensory and sympathetic neurons, suggesting that PACAP might be a mediator in the injury response in certain parts of the nervous system. However, changes in PACAP expression have not been reported in injured motor neurons, despite the significant problem of motor neuron degeneration in injury and in several neurological diseases. We examined here changes in gene expression of PACAP and two high-affinity PACAP receptors, PAC(1) and VPAC(2), in adult rat motor neurons after facial nerve axotomy by in situ hybridization. PACAP gene expression was very low in facial motor neurons of normal rats. However, a robust time-dependent increase in PACAP mRNA was observed in the facial motor nucleus in most or all axotomized motor neurons. This induction was detectable 6 hr after axotomy, and peaked at 48 hr, when expression on the injured side averaged more than 20-fold higher than that on the contralateral side. Thereafter, PACAP mRNA levels decreased slightly, but remained more than 10-fold elevated for as long as 30 days after axotomy. In contrast to PACAP, gene expression for both the PAC(1) and VPAC(2) receptor was high in facial motor neurons of normal rats. No significant change was observed for VPAC(2) receptor gene expression in facial motor neurons after axotomy, whereas gene expression for the PAC(1) receptor became significantly decreased. The results indicate that the PACAP ligand receptor system is tightly regulated in the facial motor nucleus after axotomy, providing evidence that PACAP may be involved in motor injury responses. PMID- 10467268 TI - Inhibition of AMPA responses by mutated presenilin 1. AB - The inheritance of Alzheimer's disease in some families, as well as ablation/rescue genetics in mice, suggest that point mutations in the presenilin 1 (PS1) gene can cause disease through an unknown gain-of-function. While mutations associated with familial Alzheimer's can alter apoptotic rates and beta amyloid precursor processing, it is possible that other physiological effects contribute to pathogenesis. We have begun to explore effects on neurotransmission by monitoring responses of the neuropotent Ntera-2 cell line expressing wild-type PS1 or a FAD mutant thereof. Although no differences were initially apparent in calcium responses of metabotropic receptors, responses to glutamate were dampened in cells expressing the L286V mutant of PS1. Analysis of ionotropic agonists demonstrated that AMPA receptor alterations were responsible for this effect, whereas NMDA responses were unaltered. These data suggest that PS1 mutation could lead to cognitive deficits through subtoxic physiological effects. PMID- 10467269 TI - Outpatient management of fever in children with sickle cell disease (SCD) in an African setting. AB - Because hospitalization and intravenous antibiotics for treatment of a potentially fatal bacterial infection in febrile children with sickle cell disease (SCD) are difficult to apply, outpatient treatment has been considered in developed countries for selected patients. Eligibility criteria and procedures may differ in developing countries because of unique economic and social conditions. After clinical evaluation within 36 hr of the onset of a fever exceeding 38.5 degrees C, children with SCD who are being closely followed as a part of a SCD cohort in Cotonou (West Africa), were treated as outpatients. The antibiotic regimen consisted of intramuscular injection of ceftriaxone 50 mg/kg/day for 2 days followed by amoxicillin 25 mg/kg x 3/day x 4 days and oral hyper-hydration. Patients were observed for 6 hr and thereafter discharged with a medical control at day 2, day 8 + day 15. All 60 children included completed their treatment, and none were lost to follow-up. A definite or a presumed bacterial infection was the cause of the febrile episode in 76.7% of cases. An appreciable decrease in fever was observed from day 2 and only 2 patients were hospitalized at day 3, one for abdominal painful crisis and one other for persistent fever without documented infection. No severe bacterial infections, recurrence of febrile episode, nor death were encountered during the follow-up. The cost of this outpatient approach is US $30 per patient as compared to US $140 per patient if the patient had been hospitalized. Outpatient management of febrile episode in children with SCD is feasible and cost-effective in Sub Saharan African. It requires, however, improved medical education on SCD and immediate medical attention after the onset of fever. PMID- 10467271 TI - Stability and sterility of a recombinant factor VIII concentrate prepared for continuous infusion administration. AB - Minipumps may facilitate cost-effective and convenient continuous infusion (CI) therapy for severe hemophilia A. This study evaluated the in vitro sterility, ability to support bacterial growth, and specific activity stability of a recombinant factor VIII (FVIII; Bioclate, Centeon) delivered by simulated CI at a variety of temperatures and after the addition of heparin or antibiotic. Closed system CIs of Bioclate (89.5 IU/ml) with and without heparin were sampled and cultured over a 6 day period. Bioclate (53.7 IU/ml) with and without heparin or vancomycin was inoculated with 102-105 CFU/ml of S. aureus, S. epidermidis, Escherichia coli, E. cloacae, or Y. enterocolitica and assessed by quantitative culture after 1 and 3 days. The stability of Bioclate (50, 100, and 250 IU/ml) at three temperatures (21 degrees C, 37 degrees C, and 39 degrees C) with and without heparin or vancomycin was tested over a period of 28 days. FVIII activity was measured in triplicate by a chromogenic assay (Coamatic Factor VIII, Chromogenix) and purity evaluated by Western blot. No bacterial growth was detected during CI of FVIII for up to 6 days. Following bacterial inoculation, there was rapid growth (>3 log increase) of all tested bacterial species except S. aureus which only displayed a 1 log expansion at 3 days. The addition of heparin containing 9.45 microg/U benzyl alcohol had no effect on bacterial growth. The addition of vancomycin caused a modest suppression of S. aureus growth but not of E. coli. Diluent alone did not support bacterial growth. Neither concentration, increased temperature, nor the addition of heparin or vancomycin had a significant effect on FVIII activity stability. Samples retained >75% baseline activity for between 3 and 7 days, except the infusion of Bioclate 50 IU/ml plus heparin maintained at 21 degrees C which remained stable for 28 days. Western blot analysis supported the activity assay findings. Standard and concentrated preparations of Bioclate are suitable for CI when delivered by the MiniMed 404-SP minipump. Because of the observed nutritive capability of this FVIII concentrate for sustaining bacterial growth, any contamination could result in systemic infection. PMID- 10467270 TI - Phenotypic variability of Filipino beta(o)-thalassemia/HbE patients in Indonesia. AB - Three Indonesian patients with identical genotypes, each compound heterozygotes for Filipino beta(o)-thalassemia/HbE, expressed different clinical severities. One patient has mild disease and is transfusion independent, while the other two are severely affected and transfusion dependent. The size of the Filipino beta(o) globin gene deletion was confirmed to be 45 kb, resolving conflicting values given in the literature. Neither ameliorating genetic factors such as alpha globin gene deletions or the XmnI restriction site polymorphism at position -158 upstream of the (G)gamma-globin gene, nor differences in beta-globin gene haplotype, explain the phenotypic variation. These observations have implications for the development of antenatal diagnosis in Indonesia, as at present it is not possible to give an accurate prediction of severity of phenotype for this common genotype. PMID- 10467272 TI - N-acetylcysteine and clotrimazole inhibit sickle erythrocyte dehydration induced by 1-chloro-2,4-dinitrobenzene. AB - Clotrimazole, a specific inhibitor of the Ca(2+) activated potassium (Gardos) channel, and the antioxidant N-acetylcysteine were found to inhibit the in vitro formation of high-density sickle cells induced by treatment with 1-chloro-2,4 dinitrobenzene (CDNB). The CDNB induced leakage of K(+) can be inhibited by treatment of SS erythrocytes with 20 mM N-acetylcysteine. We conclude that the effect of N-acetylcysteine in preventing K(+) efflux and formation of high density sickle cells is related to its ability to protect the Gardos channel from oxidative damage caused by diminished levels of reduced glutathione. This effect is due to the ability of N-acetylcysteine to maintain an appropriate level of reduced glutathione and its direct antioxidant activity. PMID- 10467273 TI - Molecular basis for Rh(null) syndrome: identification of three new missense mutations in the Rh50 glycoprotein gene. AB - Rh(null) is a rare autosomal recessive disorder characterized by an absence of Rh antigens and a varying degree of hemolytic anemia and spherostomatocytosis. We report studies of two Japanese Rh(null) cases and describe three new missense mutations of RHAG, the locus that encodes Rh50 glycoprotein and modulates Rh antigen expression. In Rh(null)(HT), RHAG harbored in exon 6 two G-->A transitions, GTT-->ATT and GGA-->AGA, which cause Val(270)-->Ile and Gly(280)- >Arg substitutions, respectively. These missense mutations were cotransmitted from the propositus to the children and were predicted to reside in endoloop 5 and transmembrane (TM) segment 9, respectively. In Rh(null)(WO), RHAG contained in exon 9 a single G-->T transversion, GGT-->GTT, which caused a Gly(380)-->Val missense change in TM12 segment. The G-->T transversion, which is located at the +1 position of exon 9, had also affected pre-mRNA splicing and caused partial exon skipping. Although both Rh(null) cases had a structurally normal RH antigen locus, hemagglutination and immunoblotting showed no expression of Rh antigens or proteins. These results correlate each mutation with a structural defect in the respective TM domain of Rh50 glycoprotein. PMID- 10467274 TI - In vitro effect on human leukemic K562 cells of co-administration of liposome associated retinoids and cytosine arabinoside (Ara-C). AB - The administration of retinoids has been demonstrated to be of potential utility in the therapy of a wide spectrum of neoplastic pathologies due to the ability to induce differentiation in a large variety of primary tumor cells as well as in vitro cultured cell lines. Moreover, a number of compounds, including hemin, cytosine arabinoside, and 5-azacytidine are able to induce erythroid differentiation of the erythroleukemic cell line K562. In this paper we determined whether a combined treatment of K562 cells with suboptimal concentrations of cytosine arabinoside and retinoids containing liposomes lead to a full expression of differentiated functions. Liposomes were prepared by reverse phase evaporation technique followed by extrusion through polycarbonate filters. Cell growth kinetics studies and intracellular detection of hemoglobin by benzidine staining were performed. The results obtained showed that the combined treatment with liposomes containing retinoids and sub-optimal concentration of ara-C is an effective strategy to induce K562 cell differentiation, minimizing at the same time toxic effects. Control experiments aimed to determine possible selection of subpopulations of K562 cells suggest that the observed results are not related to toxicity and/or potential selection of induced cells. In conclusion, liposomally delivered retinoids could be proposed for differentiation therapy as an effective strategy in the treatment and management of malignancy. In addition, the finding that liposomally delivered retinoids increase the capacity of cytosine arabinoside to induce erythroid differentiation, could be of interest in studies aimed at the development of treatment able to reactivate fetal globin genes in beta-thalassemia patients. PMID- 10467275 TI - Multicentric warfarin-induced skin necrosis complicating heparin-induced thrombocytopenia. AB - Two patients developed catastrophic multicentric skin necrosis while receiving warfarin to treat venous thromboembolism complicated by immune-mediated heparin induced thrombocytopenia (HIT). Patient 1 developed skin necrosis involving the breasts, thighs, and face, as well as venous limb gangrene and bilateral hemorrhagic necrosis of the adrenal glands, resulting in death. The second patient developed bilateral mammary necrosis necessitating mastectomies, as well as skin necrosis involving the thigh. Neither patient had an identifiable hypercoagulable syndrome, other than HIT. HIT may represent a risk factor for the development of multicentric warfarin-induced skin necrosis (WISN). PMID- 10467276 TI - Loss of maternal allele in a child with myelodysplastic syndrome and monosomy 7. AB - Monosomy 7 or partial deletion of the long arm of chromosome 7 is frequently described in children with myelodysplastic syndrome and acute myeloblastic leukemia. Parental origin of chromosome 7 in children with sporadic monosomy 7 has been examined very rarely. To investigate if monosomy 7 shows parent-of origin, we have studied a female child with monosomy 7 and de novo myelodysplastic syndrome by a series of polymorphic polymerase chain reaction markers. We found loss of maternal allele and discussed the results with the previous reports. PMID- 10467277 TI - Carbamazepine-induced thrombocytopenia defined by a challenge test. AB - Carbamazepine (CBZ), a widely used anticonvulsant, occasionally causes serious hematologic disorders. A 12-year-old boy was admitted because of a diffuse petechial rash and profound thrombocytopenia (10 x 10(9) platelets/l), after having been treated for epilepsy with CBZ for 12 days. Seven days following withdrawal of CBZ and initiation of prednisolone therapy, the platelet count recovered. In a subsequent challenge test with CBZ, platelet counts again decreased, and the levels of platelet-associated IgG and serum interleukin-6 increased. No antibodies against platelet glycoprotein IIb/IIIa or Ib were detected in plasma. We believe that this is the first reported occasion when CBZ induced thrombocytopenia has been defined by a rechallenge test. PMID- 10467278 TI - Splenectomy during partial remission in thrombotic thrombocytopenic purpura with prolonged plasma exchange dependency. AB - Some patients with thrombotic thrombocytopenic purpura (TTP) remain plasma exchange-dependent for prolonged periods of time. This exposes patients to risk, uses substantial resources, and requires prolonged hospitalization. We have splenectomized 7 such patients following 25-42 plasma exchanges while patients were in partial remission only and were clinically stable. In 6 patients, including 1 with TTP secondary to mitomycin C, thrombocytopenia promptly resolved. Relapse has not occurred during 18 or more months of observation. The seventh patient did not respond. We conclude that splenectomy should be considered as an alternative to continued plasma-exchange therapy in such patients. PMID- 10467279 TI - Spontaneous venous thrombosis in a young patient with combined factor V Leiden and lupus anticoagulant. AB - We describe a case of a 28-year-old man who developed an extensive spontaneous deep venous thrombosis. Testing revealed heterozygotic factor V Leiden mutation, and the presence of both lupus anticoagulant (LA) and elevated IgM anticardiolipin antibody (ACA). Several family members were found to be heterozygous for factor V Leiden. A paternal aunt had the factor V Leiden mutation, an elevated plasma homocysteine and a borderline increased IgG ACA level. No other family member had a history of a venous thrombotic event. This case illustrates that evaluation of young patients who present with venous thrombosis should be performed for both hereditary and acquired thrombophilic defects. The family studies suggest that the presence of a lupus anticoagulant may be more clinically significant than elevated ACA in risk assessment. Although screening family members when the proband carries factor V Leiden is controversial, psychological reassurance of those who test negative and simple advice on occupations or social habits (e.g., smoking) for those who test positive may be important benefits. PMID- 10467280 TI - Pernicious anemia (PA) subsequent to insulin-dependent diabetes mellitus and idiopathic thrombocytopenic purpura, and effects of oral cobalamin on PA. PMID- 10467281 TI - A 78-year-old man with sickle-cell anemia. PMID- 10467282 TI - Possible cyclosporin-danazol interaction in a patient with aplastic anaemia. PMID- 10467283 TI - Proton magnetization transfer of metabolites in human brain. AB - Off-resonance or pulsed on-resonance saturation pulses were used together with localized proton magnetic resonance spectroscopy in three brain regions of 20 healthy individuals. Statistically significant signal attenuations were observed for creatine-containing metabolites in posterior-parietal brain (12%), basal ganglia (18%), and cerebellum (15%). N-acetyl- and choline-containing metabolites were not significantly attenuated upon application of saturation pulses in either brain region. The findings are interpreted to reflect possible magnetization transfer between pools of creatine-containing metabolites with different molecular mobility. Magn Reson Med 42:417-420, 1999. PMID- 10467284 TI - Residual dipolar coupling of the Cr/PCr methyl resonance in resting human medial gastrocnemius muscle. AB - This paper presents a detailed analysis of the TE dependence of the creatine methyl proton signal at 3.02 ppm, resulting from a symmetric PRESS sequence applied to the resting human gastrocnemius muscle. The analysis shows that a two component decay of the central peak of the dipolar-coupled-methyl triplet should be interpreted as the superposition of a rapid ( approximately 34 msec) dipolar dephasing and a less rapid ( approximately 162 msec) transverse relaxation. These data do not support a two-pool hypothesis for TE dependence of this signal. Magn Reson Med 42:421-424, 1999. PMID- 10467285 TI - Pseudo-continuous arterial spin labeling technique for measuring CBF dynamics with high temporal resolution. AB - Cerebral blood flow (CBF) can be measured noninvasively with nuclear magnetic resonance (NMR) by using arterial water as an endogenous perfusion tracer. However, the arterial spin labeling (ASL) techniques suffer from poor temporal resolution due to the need to wait for the exchange of labeled arterial spins with tissue spins to produce contrast. In this work, a new ASL technique is introduced, which allows the measurement of CBF dynamics with high temporal and spatial resolution. This novel method was used in rats to determine the dynamics of CBF changes elicited by somatosensory stimulation with a temporal resolution of 108 ms. The onset time of the CBF response was 0.6 +/- 0.4 sec (mean +/- SD) after onset of stimulation (n = 10). The peak response was observed 4.4 +/- 3.7 sec (mean +/- SD) after stimulation began. These results are in excellent agreement with previous data obtained with invasive techniques, such as laser Doppler flowmetry and hydrogen clearance, and suggest the appropriateness of this novel technique to probe CBF dynamics in functional and pathological studies with high temporal and spatial resolution. Magn Reson Med 42:425-429, 1999. PMID- 10467286 TI - Motion-selective encoding for fast cine imaging. AB - In magnetic resonance (MR) cine imaging applications, it is common that only a fraction of the field of view is used for delineating dynamic features whereas surrounding static structures are merely served as landmarks for three dimensional orientation. This paper presents a method of using motion-selective encoding for fast cine imaging. By the use of a simple RF saturation pulse, the imaging field of view was divided into two areas, one with motion and the other without. This allowed the static material to be imaged by fast ungated acquisition and dynamic structures by reduced phase encoding. Issues related to phase error correction were addressed and the proposed algorithm was validated using images acquired from five normal subjects. The saving in imaging time was dependent on the size of the moving structures and a 35% reduction in imaging time did not cause visually noticeable changes in image quality. The application of the technique is limited to cine images with large surrounding static structures and a theoretical analysis of maximum savings in imaging time in relation to the size of the moving structures within the image field of view is provided. Magn Reson Med 42:430-435, 1999. PMID- 10467288 TI - Heteronuclear NMR studies of metabolites produced by Cryptococcus neoformans in culture media: identification of possible virulence factors. AB - The yeast, Cryptococcus neoformans var. neoformans is a major contributor to the morbidity and mortality experienced by the immunosuppressed population. With a view to providing better treatment, identification of cryptococcal virulence factors is an important goal, with most effort to date directed toward the significance of structural variations in the polysaccharide capsule. The present work describes the characterization of supernatants obtained from cryptococcal cultures. This was achieved by thorough identification of the spin systems of individual metabolites through both homonuclear and heteronuclear NMR experiments that circumvented the difficulties imposed by limited dispersion and a range of concentrations in different cultures covering more than 3 orders of magnitude. More than 30 metabolites, including amino acids, alditols, nucleosides, acetate, and ethanol, were identified by their (1)H and (13)C chemical shifts and observed long-range correlations. The possible contribution of some detected substances to the pathogenicity of Cryptococcus neoformans is discussed. Magn Reson Med 42:442 453, 1999. PMID- 10467287 TI - Analysis of periprosthetic capsular tissue from women with silicone breast implants by magic-angle spinning NMR. AB - The amount of silicone (polydimethylsiloxane [PDMS]) in capsular tissue surgically removed from women with breast implants was measured by using (29)Si and (1)H magic-angle spinning solid-state NMR spectroscopy. Twelve women having smooth surface silicone gel-filled implants, including a subject with "low-bleed" double-lumen implants, had detectable levels of PDMS ranging from 0. 05 to 9.8% silicon in wet tissue (w/w). No silicon-containing compounds other than PDMS were detected. No correlation was found between the amount of PDMS measured in the capsular tissue and the length of implantation time (Pearson correlation coefficient, r = 0. 22). The results showed no relationship between higher amounts of PDMS and capsular contracture (p = 0.74) or other symptoms (p = 0. 53). Magn Reson Med 42:436-441, 1999. PMID- 10467289 TI - A localized double-quantum filter for in vivo detection of taurine. AB - Noninvasive detection of taurine, an important amino acid involved in numerous physiological processes, by in vivo (1)H magnetic resonance (MR) spectroscopy is complicated by severe overlap of the taurine resonances with those of a number of other metabolites. Unambiguous differentiation of the taurine resonances requires spectral editing. In this study, the development of a localized spectral editing technique based on double-quantum filtering optimized for in vivo detection of taurine is described. The sequence recovers the taurine signal while substantially eliminating overlapping resonances and provides excellent three dimensional spatial localization. The performance of the sequence is demonstrated both in phantoms and in rat brain in vivo. Magn Reson Med 42:454-460, 1999. PMID- 10467290 TI - Anisotropic and restricted diffusion of water in the sciatic nerve: A (2)H double quantum-filtered NMR study. AB - The signals of water in the different compartments of rat sciatic nerve are resolved in the (2)H double-quantum-filtered NMR spectrum, due to their different quadrupolar splittings and relaxation rates. This resolution allowed the independent measurement of the water diffusion coefficients in the different compartments. The water diffusion in all three compartments, the endoneurium, the epineurium and the axon was found to be anisotropic. Parallel to the nerve fiber the average intraxonal water diffusion coefficient was 1.11 x 10(-5) cm(2)/sec, while in the perpendicular direction the diffusion is heavily restricted. The average perpendicular diffusion coefficient ranged from 0.29 x 10(-5) cm(2)/sec to 0.05 x 10(-5) cm(2)/sec for diffusion times of 7 msec and 50 msec, respectively. Assuming restricted diffusion in nonpermeable cylinders, intra axonal mean diameters of 6.0, 7.4 and 9.0 microm were obtained for nerves taken from three different rats. Magn Reson Med 42:461-466, 1999. PMID- 10467291 TI - Equilibrium transcytolemmal water-exchange kinetics in skeletal muscle in vivo. AB - It is commonly assumed that equilibrium transcytolemmal water exchange in tissue is sufficiently frequent as to be fast on any NMR time scale achievable with an extracellular contrast agent (CR) in vivo. A survey of literature values for cell membrane diffusional permeability coefficients (P) and cell sizes suggests that this should not really be so. To evaluate this issue experimentally, we used a programmed intravenous CR infusion protocol for the rat with several rate plateaus, each of which achieved an increased steady-state concentration of GdDTPA(2-) in the blood plasma. Interleaved rigorous measurements of (1)H(2)O inversion recoveries were made from arterial blood and from a region of homogeneous thigh muscle tissue throughout the CR infusion. We made careful relaxographic analyses for the blood and muscle (1)H(2)O longitudinal relaxation times. The combined data from several animals were evaluated with a two-site model for equilibrium transcytolemmal water exchange. An excellent fitting was achieved, with parameters that agreed very well with the relevant physiological properties available in the literature. The fraction of water in the extracellular space, 0.11, is quite consistent with published values, as well as with reported tissue CR concentrations when one accounts for the restriction of CR to this space. The derived average lifetime for a water molecule in the thigh muscle sarcoplasm, 1.1 +/- 0.4 sec, implies a sarcolemmal P of 13 x 10(-4) cm/sec, which is well within the range of literature values determined in vitro. Moreover, we find that because of the exchange, the (1)H(2)O longitudinal relaxation rate constant exhibits a decided nonlinear dependence on the tissue or thermodynamic (extracellular) concentration of GdDTPA(2-). The muscle system departs the fast-exchange limit at a [CR] value of <100 micromol/L. This has significant implications for the quantitative use of CRs as MRI tracers. Magn Reson Med 42:467-478, 1999. Published 1999 Wiley-Liss, Inc. PMID- 10467292 TI - Global HDO uptake in human glioma xenografts is related to the perfused capillary distribution. AB - The aim of this study is to evaluate the existence of a possible relationship between global deuterium-labeled water (HDO) uptake rates and the diffusion geometry of human glioma xenografts in nude mice. HDO diffusion times in the whole extravascular tumor volume were estimated by combining quantitative (1)H-MR diffusion imaging and morphometric analysis of intercapillary distances in two tumor lines with a different perfused vascular architecture. HDO uptake was measured independently using (2)H-magnetic resonance spectroscopy. Time constants of HDO-uptake curves (tau) were compared to estimations of maximum HDO diffusion times (t(difmax)). Tumors with a homogeneously perfused capillary distribution showed a mono-exponential HDO uptake. The t(difmax) was comparable to tau values of HDO uptake curves: t(difmax) varied between 74 and 368 sec and the range of tau values was 115-370 sec. Heterogeneously perfused tumors had a bi-exponential HDO uptake with t(difmax) in between the tau values of the fast and slow uptake phase. These findings indicate that the global HDO uptake is related to the perfused capillary distribution in human glioma xenografts. That HDO uptake rates indeed can depend on the perfused capillary distribution was substantiated in experiments with two-dimensional (2D) models. In these models with a diffusion limited HDO uptake, HDO uptake curves could be approximated by curves derived from 2D HDO diffusion simulations. Magn Reson Med 42:479-489, 1999. PMID- 10467293 TI - Effect of carbogen breathing on the physiological profile of human glioma xenografts. AB - The aim of this study was to evaluate the effect of carbogen breathing on the physiological profile of human glioma xenografts. Near infrared spectroscopy was used to investigate changes in oxy- and deoxyhemoglobin concentrations in tumor blood. Oxygen tension changes in tumor tissue were evaluated by (19)F-MR relaxometry, using perfluoro-15-crown-5-ether, and modifications of tumor blood perfusion (TBP) were analyzed by fast dynamic (1)H-MR imaging of Gd-DTPA uptake. Finally, changes of the bioenergetic status and pH of tumor cells were analyzed by (31)P-MRS. After 5 to 8 min of carbogen breathing, the average oxygen tension increase in tumor tissue was 4.6 +/- 1.3 mm Hg, which is in agreement with an increase of the oxyhemoglobin concentration in tumor blood (Delta[O(2)Hb] = 9. 2 +/- 3 microM). However, simultaneously the TBP was reduced, the bioenergetic status was diminished, and pH was decreased. As 100% O(2) breathing alone did not result in a detectable increase of oxyhemoglobin in tumor blood, the increase of the tumor oxygenation by carbogen appears to be mediated by its CO(2) content. This component may cause a nutrient-limited decrease of oxidative energy metabolism, indirectly via a steal-effect and/or by inhibition of the glycolytic rate resulting from tissue acidification. Magn Reson Med 42:490-499, 1999. PMID- 10467294 TI - Perfusion-corrected mapping of cardiac regional blood volume in rats in vivo. AB - Measurement of regional blood volume (RBV) in the myocardium in vivo is important for the assessment of tissue viability and function. The method in this work is based on the acquisition of a T(1) map before and after intravascular contrast agent application. It is known that this method is influenced by perfusion that causes an overestimation of RBV values. In order to solve this problem, the new method is proposed which acquires T(1) maps with slice selective inversion pulses. Due to blood flow nonexcited spins enter the detection slice, which leads to an acceleration of the relaxation time. A model that divides tissue into two compartments is adapted to slice selective inversion in order to derive a simple expression for perfusion-corrected RBV. The aim of the study is to demonstrate the feasibility and accuracy of this technique for quantification of RBV in rat myocardium in vivo. RBV maps were obtained for five rats, and the reproducibility was determined by repeating the experiment several times. A mean RBV value of 12.8 +/- 0.7% (v/v) over all animals was obtained in the myocardium. The results were compared with RBV maps obtained with perfusion-sensitive RBV imaging in the same five rats and with first-pass RBV studies. In order to demonstrate the strength of the new method the vasodilator adenosine was administered and alterations in microcirculation were imaged. Magn Reson Med 42:500-506, 1999. PMID- 10467295 TI - Rapid imaging of hyperpolarized gas using EPI. AB - Rapid repetitive MRI of hyperpolarized (HP) gases using echo-planar imaging (EPI) has been theoretically investigated and experimentally demonstrated for (3)He in human lung. A quantitative treatment of signal attenuation and magnetization consumption for the unique circumstance of a rapidly diffusing nonrenewable magnetization source has been performed. Rapid (compared to the human respiratory cycle) and repetitive imaging of the lung gas space with EPI and a single delivered bolus of HP-(3)He is feasible using low flip angles, provided the voxels are not too small. A coarse-grid (32 x 64) EPI pulse sequence has been developed and implemented to image the lungs of healthy volunteers during rebreathing of a HP-(3)He/N(2) gas mixture. A set of three 10-mm axial slices was imaged every 0.12 sec for the 36 sec duration of rebreathing, yielding a real time visualization of ventilation. Despite some mild artifacts, the images are of good quality and show changes in gas density related to respiratory physiology. Magn Reson Med 42:507-514, 1999. PMID- 10467296 TI - Optimal strategies for measuring diffusion in anisotropic systems by magnetic resonance imaging. AB - The optimization of acquisition parameters for precise measurement of diffusion in anisotropic systems is described. First, an algorithm is presented that minimizes the bias inherent in making measurements with a fixed set of gradient vector directions by spreading out measurements in 3-dimensional gradient vector space. Next, it is shown how the set of b-matrices and echo time can be optimized for estimating the diffusion tensor and its scalar invariants. The standard deviation in the estimate of the tensor trace in a water phantom was reduced by more than 40% and the artefactual anisotropy was reduced by more than 60% when using the optimized scheme compared with a more conventional scheme for the same scan time, and marked improvements are demonstrated in the human brain with the optimized sequences. Use of these optimal schemes results in reduced scan times, increased precision, or improved resolution in diffusion tensor images. Magn Reson Med 42:515-525, 1999. PMID- 10467297 TI - Color schemes to represent the orientation of anisotropic tissues from diffusion tensor data: application to white matter fiber tract mapping in the human brain. AB - This paper investigates the use of color to represent the directional information contained in the diffusion tensor. Ideally, one wants to take into account both the properties of human color vision and of the given display hardware to produce a representation in which differences in the orientation of anisotropic structures are proportional to the perceived differences in color. It is argued here that such a goal cannot be achieved in general and therefore, empirical or heuristic schemes, which avoid some of the common artifacts of previously proposed approaches, are implemented. Directionally encoded color (DEC) maps of the human brain obtained using these schemes clearly show the main association, projection, and commissural white matter pathways. In the brainstem, motor and sensory pathways are easily identified and can be differentiated from the transverse pontine fibers and the cerebellar peduncles. DEC maps obtained from diffusion tensor imaging data provide a simple and effective way to visualize fiber direction, useful for investigating the structural anatomy of different organs. Magn Reson Med 42:526-540, 1999. PMID- 10467298 TI - Rapid iterative reconstruction for echo planar imaging. AB - A rapid automated method for reconstructing echo planar imaging (EPI) data has been developed and is shown to improve image quality by suppressing the troublesome ghost artifact. The algorithm can be applied without prior knowledge obtained from either reference scans or operator intervention. It first estimates, then improves iteratively, the parameters for a linear phase correction applied directly to the complex image data derived from odd and even echoes. The theory used to derive the criteria employed in the iteration provides insight into mechanisms that allow the process to work. Magn Reson Med 42:541 547, 1999. PMID- 10467299 TI - A computationally efficient method for tracking reference position displacements for motion compensation in magnetic resonance imaging. AB - A fast and computationally efficient method for detecting and tracking the displacement of a reference structure within the body using MR imaging is described. This method is used to determine the position of the diaphragm in order to synchronize the data acquisition to the same relative position of the abdominal and thoracic organs, thereby minimizing or eliminating respiratory motion artifacts. The method described uses the time domain linear phase shift of a reference structure to determine its spatial positional displacement as a function of the respiratory cycle. The signal from a two-dimensional rectangular excitation column is first Fourier-transformed to the image domain, apodized, and then transformed back to the time domain. The relative displacement of a target edge in the image domain is determined from an autocorrelation of the resulting time domain information. This technique was found to require between three and eight times less computation than either cross-correlation or least-squares analysis, depending on the navigator parameters. Magn Reson Med 42:548-553, 1999. PMID- 10467300 TI - Improved device definition in interventional magnetic resonance imaging using a rotated stripes keyhole acquisition. AB - Keyhole acquisition techniques have been used to reduce image acquisition times primarily in contrast agent studies and via simulation in interventional MRI procedures. More recent simulations have suggested that improved definition of an interventional device [e.g., biopsy needles, radio frequency (RF) electrodes] could be achieved by rotating the keyhole pattern in k-space so that the read out direction lies perpendicular to the device orientation in real space. This study seeks to validate the earlier predictions of improved efficacy of a rotated stripes keyhole acquisition in actual in vitro and in vivo interventional MR imaging procedures. A true-FISP sequence was modified to perform central stripes keyhole (as known as conventional keyhole) acquisitions after a full initial reference data set was acquired. The gradients of this sequence were then modified to rotate the k-space definition and the keyhole stripes by 10 degrees, 20 degrees, 30 degrees, 45 degrees, and 60 degrees from their conventional k space orientation. Acquisitions were performed during insertion of interventional devices in phantom and in vivo RF ablation procedures, using the modified sequence selected which placed the phase encoding axis at parallel and perpendicular orientations to the devices. Resulting images were compared between the two orientations for needle width and tip accuracy. Apparent needle width was thinner and tip position more accurately determined for placement of phase encoding parallel to the needle in all cases. Rotated keyhole imaging provides the required temporal advantage of conventional keyhole imaging along with a near optimal definition of an interventional device when the phase encoding is oriented parallel to the direction of the needle motion. Magn Reson Med 42:554 560, 1999. PMID- 10467301 TI - Modular gradient coil: A new concept in high-performance whole-body gradient coil design. AB - A new concept in high-performance MR gradient coil design is presented which we have called the Modular Gradient Coil (MGC). This novel design approach results in an actively shielded whole-body gradient coil containing multiple and independent elements, integrated onto a single former, for generating gradient fields along each of the three axes (x, y, and z). These elements can be energized in a number of configurations, using a single gradient power supply unit (PSU), to generate a whole range of gradient performance levels. The design criteria for the MGC also include a requirement to prohibit peripheral nerve stimulation in all of its modes of operation. This requirement is achieved, while simultaneously providing high performance, by specifying different volumes of gradient linearity for each of the operating modes. Magn Reson Med 42:561-570, 1999. PMID- 10467302 TI - BOLD MRI of the human cervical spinal cord at 3 tesla. AB - The feasibility of functional MRI of the spinal cord was investigated by carrying out blood oxygen-level dependent (BOLD) imaging of the human cervical spinal cord at a field of 3 T. BOLD imaging of the cervical spinal cord showed an average intensity increase of 7.0% during repeated exercise with the dominant hand with a return to baseline during rest periods. The areas of activation were predominantly on the same side of the spinal cord as the hand performing the exercise, between the levels of the sixth cervical and first thoracic spinal cord segments. The direct correspondence between these areas and those involved with the transmission of motor impulses to the hand, and reception of sensory information from the hand, demonstrates that spinal functional magnetic resonance imaging is feasible. Magn Reson Med 42:571-576, 1999. PMID- 10467303 TI - Method for improved multiband excitation profiles using the Shinnar-Le Roux transform. AB - A method to design multiband RF pulses for magnetic resonance imaging is described. The method is based on the Shinnar-Le Roux transform and involves a phase correction that provides control over the phase of the excited bands. The theory behind the method and this phase correction is outlined. The method is demonstrated with the design of RF pulses for Hadamard encoding and Haar wavelet encoding. Experimentally measured excitation profiles and images for RF pulses designed with the new method are compared to those designed by the conventional method. The conventional method is shown to result in distortion of the excitation profile when the bands are closely spaced. A 78% reduction in this distortion is attributed to the new method. This translates into a 52% reduction of out-of-slice signal in Haar wavelet encoding. Magn Reson Med 42:577-584, 1999. PMID- 10467304 TI - Geometric distortion correction in gradient-echo imaging by use of dynamic time warping. AB - Distortion caused by local static field variations in magnetic resonance imaging can be corrected by using a method developed by Chang and Fitzpatrick (CF) that requires two acquisitions identical except for the polarity of the read gradient. This approach has previously been applied to spin-echo imaging. In this report, its role in correcting geometric distortion in gradient-echo imaging is investigated using a dynamic programming algorithm. This approach appears to successfully handle many of the edge artifacts otherwise inherent in the CF method. Once the correction has been made, it is then possible to extract the local magnetic field itself. Magn Reson Med 42:585-590, 1999. PMID- 10467305 TI - Vascular filters of functional MRI: spatial localization using BOLD and CBV contrast. AB - The spatial distributions of functional activation of rat somatosensory cortex by forepaw stimulation were quantitatively compared using blood oxygen level dependent (BOLD) signal and signal weighted by cerebral blood volume (CBV). The BOLD contrast to noise (CNR) distribution showed a significant dorsal shift with respect to the CBV method at fields strengths of 2 T (0.69 +/- 0.09 mm) and 4.7 T (0.44 +/- 0.15 mm). These shifts were attributed to the gradient of resting state blood volume across somatosensory cortex and the different CNR characteristics of the two image methods. The underlying principles suggest that the CBV method has a more uniform sensitivity to percent changes in functional indicators (blood volume or deoxygenated hemoglobin) across regions of variable resting state CBV. Magn Reson Med 42:591-598, 1999. PMID- 10467306 TI - Ex vivo EPR detection of nitric oxide in brain tissue. AB - The concentration of nitric oxide (NO) was measured in the brain of septic-shock animals by electron paramagnetic resonance spectrometry (EPR). NO was spin trapped and quantitated in several regions of the brain (cortex, hippocampus, hypothalamus, cerebellum, and olfactory bulb) as well as other organs (liver, kidney, and heart) of rats induced with lipopolysaccharide (LPS) using Fe(II)/dithiocarbamate complexes containing diethyldithiocarbamate (DETC) or N methyl-D-glucamine (MGD). The spin trap, (DETC)(2)-Fe(II), complexed NO generated in all tissues examined, but (MGD)(2)-Fe(II) complex was ineffective in detecting NO in the brain of septic-shock rats, although identical amounts of NO were detected in the liver with either spin trap. A triplet EPR spectrum of (DETC)(2) Fe(II)-NO with a(N) = 12.8 gauss and g = 2.04 was observed in the cortex, hippocampus, hypothalamus, cerebellum, but not the olfactory bulb. The amount of NO in the brain was about 20% of that found in the liver. The (DETC)(2)-Fe(II)-NO signal in all the tissues of septic-shock rats was markedly suppressed by preadministration of the nitric oxide synthase (NOS) inhibitors, N(G)-monomethyl L-arginine (L-NMMA) or 3-bromo-7-nitroindazole, suggesting that the NO detected from brain tissue was produced enzymatically by NOS. In contrast to previous studies on the liver and other organs, phenyl-N-tert-butyl nitrone (PBN), did not suppress iNOS expression in brain tissue of LPS-treated rats. This could be due to a totally different regulation system for iNOS in liver versus brain tissue. Magn Reson Med 42:599-602, 1999. PMID- 10467307 TI - MRI measurement of cell volume fraction in the perfused rat hippocampal slice. AB - T(1)-weighted NMR imaging of the isolated perfused rat hippo-campal slice was used to estimate cell volume fraction. Eight brain slices were studied in artificial cerebrospinal fluid (aCSF) using a 600 MHz narrow bore spectrometer and a home built perfusion chamber. Cell volume fraction was calculated as 1 - f(ECS), where f(ECS) is the distribution volume of gadodiamide in the slice. This was determined by measuring the T(1) of the slice before and after perfusion with gadodiamde. A mean cell volume fraction of 0.66 +/- 0. 04 was estimated. The addition of 60 mM mannitol to three of the brain slices produced a 26% decrease in the cell volume fraction. The technique affords a simple means of estimating cell volume fraction and can be extended to produce images reflecting cell density. Magn Reson Med 42:603-607, 1999. PMID- 10467308 TI - Assessment of cerebral oxidative metabolism with breath holding and fMRI. AB - Carbon dioxide inhalation can be used to map changes in cerebral metabolic rate of oxygen (CMRO(2)) during neuronal activation with functional MRI (fMRI). A hypercapnic stress also can be achieved with a simple breath-holding test. Using this test as means of manipulating cerebral blood flow (CBF) independent of CMRO(2), we assessed changes in CMRO(2) during visual stimulation. With this task, CBF increased by 61 +/- 7%, whereas CMRO(2) changed by 2.43 +/- 4.97%. These results are in good agreement with previous positron emission tomographic (PET) data, indicating that changes in oxidative metabolism during focal neuronal activity can potentially be determined with the breath-holding test. This test could easily be performed during a routine MRI examination. Magn Reson Med 42:608 611, 1999. PMID- 10467310 TI - Response to "Optimal partial volume estimation in MRI" PMID- 10467309 TI - Optimal partial volume estimation in MRI. PMID- 10467312 TI - Prediction of chiral separations using a combination of experimental design and artificial neural networks. AB - In this work the advantages of using artificial neural networks (ANNs) combined with experimental design (ED) to optimize the separation of amino acids enantiomers, with alpha-cyclodextrin as chiral selector, were demonstrated. The results obtained with the ED-ANN approach were compared with those of either the partial least-squares (PLS) method or the response surface methodology where experimental design and the regression equation were used. The ANN approach is quite general, no explicit model is needed, and the amount of experimental work can be decreased considerably. PMID- 10467311 TI - Stereoselective drug release from ketoprofen and ricobendazole matrix tablets. AB - Crystalline characteristics of racemic, pure R and S enantiomers and physical mixtures of Ketoprofen (KET) have been studied by DSC and X-ray diffractometry. Aqueous solubilities were 182.6 +/- 9.1 microg/ml for racemic KET, 259.6 +/- 6.6 microg/ml for R-KET, and 304.3 +/- 2.7 microg/ml for S-KET. Matrix tablets made with racemic and physical mixtures of KET show stereoselective drug release, which is faster for S-KET than for R-KET. This effect is more marked when the chiral excipient hydroxypropylmethylcellulose (HPMC) is used in place of the achiral Eudragit RL. Stereoselectivity of release is also affected by the amount of KET. Similar results were obtained when another chiral drug with low solubility, Ricobendazole (RBZ), is used. Depending on the excipient and drug dosage, more or less marked stereoselective drug release is obtained in RBZ matrix tablet formulations. PMID- 10467313 TI - Enantiomeric separation of N-protected amino acids by non-aqueous capillary electrophoresis using quinine or tert-butyl carbamoylated quinine as chiral additive. AB - A capillary electrophoretic (CE) method for the enantioseparation of N-protected chiral amino acids was developed using quinine and tert-butyl carbamoylated quinine as chiral selectors added to nonaqueous electrolyte solutions (NACE). A series of various N-derivatized amino acids were tested as chiral selectands, and in order to optimize the CE enantioseparation of these compounds, different parameters were investigated: the nature of the organic solvent, the combination of different solvents, the nature and the concentration of the background electrolyte, the selector concentration, the capillary temperature, and the applied voltage. The influence of these factors on the separation of the analyte enantiomers and the electroosmotic flow was studied. Generally, with tert-butyl carbamoylated quinine as chiral selector, better enantioseparations were achieved than with unmodified quinine. Optimum experimental conditions were found with a buffer made of 12. 5 mM ammonia, 100 mM octanoic acid, and 10 mM tert-butyl carbamoylated quinine in an ethanol-methanol mixture (60:40 v/v). Under these conditions, DNB-Leu enantiomers could be separated with a selectivity factor (alpha) of 1.572 and a resolution (Rs) of 64.3; a plate number (N) of 127,000 and an asymmetry factor (As) of 0.93 were obtained for the first migrating enantiomer. PMID- 10467314 TI - S-perindopril assay using a potentiometric, enantioselective membrane electrode. AB - A potentiometric, enantioselective membrane electrode based on graphite paste (graphite powder and paraffin oil) has been constructed. The graphite paste is impregnated with a 10(-3) mol/L 2-hydroxy-3-trimethylammoniopropyl-beta cyclodextrin (as chloride salt) solution. The potentiometric, enantioselective membrane electrode can be used reliably for enantiopurity tests of S-perindopril using a chronopotentiometric (zero current) technique, in the 10(-5)-10(-2) mol/L concentration range (detection limit 5 x 10(-6) mol/L), with an average recovery of 99.58% (RSD = 0.33%). The enantioselectivity was determined over R-perindopril and D-proline. The response characteristics of the enantioselective, potentiometric membrane electrode were also determined for R-perindopril. It was shown that L-proline is the main interfering compound. The surface of the electrode can be regenerated simply by polishing, obtaining a fresh surface ready to be used in a new assay. PMID- 10467315 TI - Chiral recognition of verapamil by cyclodextrins studied with capillary electrophoresis, NMR spectroscopy, and electrospray ionization mass spectrometry. AB - Capillary electrophoresis (CE) allows the observation of the opposite affinities of the enantiomers of (+/-)-verapamil [2-isopropyl-2,8-bis(3,4-dimethoxyphenyl)-6 methyl-6-azaoctannitrile+ ++, VP] toward beta-cyclodextrin (beta-CD) and heptakis(2,3, 6-tri-O-methyl)-beta-CD (TM-beta-CD). In addition, in the presence of beta-CD in the background electrolyte, longer migration times and lower separation factors were observed compared to TM-beta-CD. The binding constants of (+)- and (-)-VP with beta-CD and TM-beta-CD determined using (13)C NMR spectroscopy explain the results observed in CE. Electrospray ionization mass spectrometry (ESI-MS) was used as an alternative technique for the characterization of VP-CD complexes. PMID- 10467316 TI - Enantioselective synthesis and teratogenicity of propylisopropyl acetamide, a CNS active chiral amide analogue of valproic acid. AB - Propylisopropyl acetamide (PID), an amide analogue of the major antiepileptic drug valproic acid (VPA), possesses favorable anticonvulsant and CNS properties. PID contains one chiral carbon atom and therefore exists in two enantiomeric forms. The purpose of this work was to synthesize the two PID enantiomers and evaluate their enantiospecific teratogenicity. Enantioselective synthesis of PID enantiomers was achieved by coupling valeroyl chloride with optically pure (4S)- and (4R)-benzyl-2-oxazolidinone chiral auxiliaries. The two oxazolidinone enolates were alkylated with isopropyl triflate, hydrolyzed, and amidated to yield (2R)- and (2S)-PID. These two PID enantiomers were obtained with excellent enantiomeric purity, exceeding 99.4%. Unlike VPA, both (2R)- and (2S)-PID failed to exert teratogenic effects in NMRI mice following a single 3 mmol/kg subcutaneous injection. From this study we can conclude that individual PID enantiomers do not demonstrate stereoselective teratogenicity in NMRI mice. Due to its better anticonvulsant activity than VPA and lack of teratogenicity, PID (in a stereospecific or racemic form) has the potential to become a new antiepileptic and CNS drug. PMID- 10467317 TI - Ring inversion barrier of diazepam and derivatives: An ab initio study. AB - Systematic ab initio calculations were performed to investigate the ring inversion process of various 1,4-diazepines including diazepam, N(1) desmethyldiazepam, and 3-methyl-N(1)-desmethyldiazepam. The diazepine ring adopts a shape of a boat; owing to asymmetric substitution two such boats are possible in mirror image relation to each other. In the present study both structural and solvent effects were investigated on the energetics of ring inversion of nine diazepine derivatives. The calculated ring inversion barriers for diazepam (17.6 kcal/mol) and N(1)-desmethyldiazepam (10.9 kcal/mol) are in good agreement with the corresponding experimental data. In the cases of diazepam and N(1) desmethyldiazepam, the calculated minimum energy path of the ring inversion is asymmetric contrary to the fact that the terminals (M and P conformers) are equienergetic. PMID- 10467318 TI - Determination of absolute configuration in 4-aryl-3, 4-dihydro-2(1H)-pyrimidones by high performance liquid chromatography and CD spectroscopy. AB - The absolute configuration of three 4-aryl-3, 4-dihydro-2(1H)-pyrimidones (Biginelli compounds, DHPMs) was established by comparison of the typical circular dichroism (CD) spectra of individual enantiomers with reference samples of known absolute configuration. The enantiomers were obtained by semipreparative separation of racemic mixtures on a Chiralcel OD-H chiral stationary phase. The method was used to establish the enantiopreference of various lipases in biocatalytic kinetic resolution experiments employing activated DHPM esters. PMID- 10467319 TI - Enantioseparations with the macrocyclic antibiotic ristocetin A using a countercurrent process in CE. AB - The chiral selectivity of ristocetin A was examined in a countercurrent process in CE using a coated column to suppress electroosmotic flow. Excellent enantioseparations of several nonsteroidal antiinflammatories, dansylamino acids, dinitrophenyl-derivatized amino acids, and other optically active compounds were achieved. The chiral selectivity of ristocetin A also was examined as a function of antibiotic concentration and pH. Enantioresolution was found to significantly improve with a slight increase in migration time upon increasing chiral selector concentration. Enantioselectivities were found to be greatly influenced by pH of the running buffer. PMID- 10467320 TI - Evaluation of the enantiomeric composition of amino acids in tobacco. AB - Despite the fact that several studies have reported the concentrations of various free amino acids in tobacco, their enantiomeric composition is unknown. Both the absolute and enantiomeric compositions of proline, alanine, asparagine, aspartic acid, valine, methionine, leucine,and phenylalanine were determined for three strains of tobacco leaf, three types of smokeless tobacco, and six different blended filtered and nonfiltered reference cigarettes. Some of the highest levels of D-amino acids ever found in agricultural products were observed. Possible mechanisms for the production of these D-amino acids are considered. The relevance of D-amino acids in tobacco is discussed. PMID- 10467322 TI - Peripheral blood vs bone marrow as a source for allogeneic hematopoietic stem cell transplantation. AB - In this randomized prospective study, we included 30 patients with different hematological diseases (acute myeloid leukemia, acute lymphoblastic leukemia, chronic myeloid leukemia, myelodysplastic syndrome or severe aplastic anemia) to compare peripheral blood stem cells (PBSC) (15 patients; mean age 23) and bone marrow (BM) (15 patients; mean age 21.8) as a source for allogeneic transplantation regarding the tempo of hematopoietic recovery and the incidence of acute graft-versus-host disease (GVHD). In the BM group, the median nucleated cell count harvested was 1.3 x 10(10), while in the PBSC group, the aphereses contained a median of 4.4 x 10(6) CD34+/kg recipient weight. PBSC transplantation (PBSCT) was associated with faster hematopoietic reconstitution measured as absolute neutrophil count (ANC) >0.5 x 10(9)/l (log-rank P value <0.0018) and platelet count >25 x 10(9)/l (log-rank P value <0.0098). Seven patients (46.7%) in the BM group vs only one patient (6.7%) in the PBSC group developed acute GVHD (P = 0.013). Therefore, we conclude that PBSCT is associated with faster hematopoietic recovery and the incidence of acute GVHD does not exceed that seen with BMT. PMID- 10467321 TI - Human herpes virus 8 (HHV8) serology in allogeneic bone marrow transplant recipients. AB - Human herpes virus 8 (HHV8) may be sexually transmitted, but transmission via blood cells has not yet been excluded. We used a modified immunofluorescence assay to detect Ab to HHV8 latency-associated nuclear Ag in sera of 200 allogeneic BMT recipients and their related donors. In control subjects, Ab were found in 85% of patients with AIDS-related Kaposi sarcoma (n = 52), 34% of HIV-1 infected subjects without Kaposi sarcoma (n = 56) and 9. 5% of blood donors (n = 42). Among BMT donors, 14.5% were HHV8+, while 10% of recipients were positive before, and 18% after BMT. In the 176 HHV8-negative recipients at BMT, there was no relationship between post-BMT seroconversion, which occurred in 26 cases (15%), and the donor's serological status. Of note, 10 HHV8+ recipients before BMT became negative post-BMT. Outcome of BMT was not influenced by prior HHV8 seropositivity, seroconversion or seroreversion of recipients. That HHV8 seropositivity among blood donors from the Paris area was comparable to that of BMT donors and recipients before BMT indicates that these patients had not been at risk of HHV8 by blood products received before BMT, although post-BMT HHV8 seroconversion probably corresponded to contamination by blood transfusions rather than by the BMT. PMID- 10467323 TI - Long-term liver function of recipients with hepatitis G virus infection after bone marrow transplantation. AB - To clarify the role of hepatitis G virus (HGV) infection in liver dysfunction following allogeneic BMT, we examined cryopreserved serum samples from 33 patients who had a history of blood transfusions before BMT and whose serum samples had been stored periodically, before BMT, on day 100, and thereafter for the presence of HGV-RNA and hepatitis C virus (HCV)-RNA by reverse transcription polymerase chain reaction. Nineteen patients (58%) out of 33 were positive for HGV-RNA before BMT and 10 for HCV-RNA. All patients positive for HCV-RNA were also positive for HGV-RNA. Patients were divided into three groups according to their viral status before BMT; namely, the G+C+ group (n = 10), the G+C- group (n = 9) and the G-C- group (n = 14). Two patients in the G-C- group became positive for HGV-RNA after BMT. One patient in the G+C- group suffered an acute exacerbation of hepatitis, with GPT levels reaching over 1000 IU/l, 2 and 3 years after BMT, showing quite a different clinical course from those in the G+C- group. Excluding these three patients, GPT levels of the patients in the G+C+ group were significantly higher after day 100 and remained higher than those of patients in the G+C- and G-C- groups for at least 4 years. There were no significant differences in post-transplant GPT levels between the G+C- group and the G-C- group at any time point. Of the seven patients followed-up for 5 to 10 years, three patients became HGV-RNA-negative, while four remained positive. In the absence of HCV co-infection, the behavior of GPT values post transplant in patients with HGV infection did not differ from those without HGV infection. With respect to the patient who was G+C- and showed high values of GPT 2 and 3 years post transplant, we suspect that his liver dysfunction might have been caused by some unknown virus or etiology. PMID- 10467324 TI - Autotransplants in men with breast cancer. ABMTR Breast Cancer Working Committee. Autologous Blood and Marrow Transplant Registry. AB - The purpose of this study was to determine the outcome of high-dose therapy with autologous hematopoietic stem cell support (autotransplants) in men with breast cancer. We studied 13 men receiving autotransplants for breast cancer and reported to the Autologous Blood and Marrow Transplant Registry (ABMTR) by 10 centers. Six men had stage 2 breast cancer, four had stage 3, and three had metastatic breast cancer. Of twelve tumors tested, all were estrogen receptor positive. Median age at transplant was 50 years. The most common conditioning regimen was cyclophosphamide, thiotepa and carboplatin (n = 5); the remaining eight men received other alkylator-based regimens. Three men received bone marrow, eight received blood stem cells, and two received both for hematopoietic support. All patients had hematopoietic recovery. There were no unexpected regimen-related toxicities. Of 10 men receiving autotransplants as adjuvant therapy, three relapsed 3, 5 and 50 months post-transplant and died 16, 19 and 67 months post-transplant. Seven of 10 are disease-free with median follow-up of 23 months (range 6-50 months). Of three men treated for metastatic breast cancer, one had progressive disease and two recurrent disease at 6, 7 and 16 months post transplant. In conclusion, results of autotransplants for male breast cancer appear similar to those reported for women receiving autotransplants for breast cancer. PMID- 10467325 TI - A comparison of CD34+ cell selected and unselected autologous peripheral blood stem cell transplantation for multiple myeloma: a case controlled analysis. AB - Following ASCT for multiple myeloma, it is unclear whether relapse is due solely to the presence of residual myeloma cells after myeloablation, or whether it is in part attributable to contamination of the stem cell harvest with viable malignant cells. Positive selection of CD34+ cells markedly reduces plasma cell contamination. We performed a case controlled analysis in which 15 patients with myeloma who underwent autologous PBSCT with CD34+cell selection using the Ceprate System (index group), were compared with 15 matched controls. All subjects received an identical preparative regimen. The median times to neutrophils >/=0.5 x 10(9)/l and unsupported platelets >/=50 x 10(9)/l were 14 and 23 days for the CD34+cell selected group and 11 (P = 0.03) and 14 (P = 0.029) for the case controls. Median follow-up of purged patients from autologous PBSCT was 32 months (range 18-43). At 36 months, the probability of PFS was 47 +/- 14% and 46 +/- 14% in the index and control groups (P = 0.44). The 3 year probability of OS was 69 +/- 13% for the CD34+ cell selected arm and 66 +/- 12.4% in unpurged patients (P = 0.91). Median PFS for the cell selected group is 24 months (CI 19.1-36.0), and 29 months for controls (CI 7.1-50.9). Eleven patients undergoing cell selection remain alive, seven of whom are progression free. At the same time-point after unpurged autologous PBSCT, the corresponding figures are 12 patients alive, with seven remaining progression free. Autologous PBSCT with CD34+ cell selection is both feasible and safe, but results in delayed engraftment as compared to case controls. The 3 year probability of PFS and OS in the cell selected arm was similar to that of the unpurged controls. Our findings indicate that autologous PBSCT with CD34+ cell selection appears not to have any favourable effect on disease progression. However, the results of this case controlled analysis should be cautiously interpreted, and the role of CD34+ selection in autologous PBSCT should be further investigated by large randomised trials. PMID- 10467326 TI - Expression of C-erbB-2/HER-2 in patients with metastatic breast cancer undergoing high-dose chemotherapy and autologous blood stem cell support. AB - C-erbB-2/HER-2 (designated HER-2) is overexpressed in both primary and metastatic breast cancer and predicts poor prognosis. We investigated the expression of HER 2 in patients with metastatic breast cancer undergoing high-dose chemotherapy (HDCT) with autologous blood stem cell (ABSC) support and correlated the presence (positive) or absence (negative) of HER-2 overexpression in these patients with response to treatment, progression-free survival (PFS) and overall survival (OS). The level of HER-2 expression was analyzed in 57 patients with metastatic breast cancer undergoing HDCT with ABSC support. Plasma from peripheral blood was taken at three different time points during the course of treatment and was analyzed using an enzyme immunoassay (EIA) to detect circulating levels of the extracellular portion of HER-2. HER-2 levels were elevated (>0.2 U/mg protein) in 27/57 (47.4%) patients at one or more time points during treatment. The level of HER-2 varied during the course of treatment. Following induction chemotherapy (ICT), five patients who were negative initially, showed overexpression of HER-2. Three patients overexpressed HER-2 only after HDCT/ABSC. Response to treatment was similar in patients independent of plasma HER-2 levels. Overexpression of HER 2 was associated with a significantly shorter PFS (P = 0.004, log rank) and OS (P = 0.003, log rank) after HDCT/ABSC. HER-2 overexpression, patient age, estrogen receptor status, progesterone receptor status, and previous hormone treatment were assessed by univariate and multivariate analysis. Univariate analysis determined that only HER-2 overexpression correlated significantly with decreases in progression free survival (P = 0.005, Cox regression). Decreased overall survival correlated significantly with HER-2 overexpression (P = 0.004) and decreased expression of both estrogen receptor (P = 0.032) and progesterone receptor (P = 0.039). In multivariate analysis of these variables, only HER-2 expression levels proved to be of independent statistical significance in predicting outcome for both PFS (P = 0.007) and OS (P = 0.002). These results suggest that overexpression of HER-2, measured by EIA in plasma may predict a shorter PFS and OS in patients with metastatic breast cancer treated with HDCT and ABSC support. PMID- 10467327 TI - Red cell salvage and reinfusion in pediatric bone marrow donors. AB - We evaluated the use of a semi-automated processing technique to salvage red blood cells from pediatric bone marrow donors to minimize the risk of severe anemia following bone marrow harvest and ABO incompatibility in the recipient. Sixty healthy, HLA-matched, pediatric donors of bone marrow hematopoietic cells with a median age 8.0 years (2-19) were studied. Thirteen of the donor-recipient pairs were ABO incompatible. There were 60 recipients with a median age of 8.6 years (2 months to 20.8 years). Bone marrow was harvested under general anesthesia, filtered in the operating room and then transferred to the stem cell laboratory for processing. Samples were obtained for cell count, CD34+ quantification, colony assay, viability, and bacteriologic cultures before and after processing. The cells were processed in a semi-automated closed system (Stericel, Terumo) by density gradient separation with Ficoll-Hypaque and then washed. Two aliquots were obtained: one containing the mononuclear cell layer to be infused to the recipient and the other the washed red cells to be infused to the donor. The median volume harvested was 608 +/- 40.42 ml (278-1409), while the final volume infused was 174 +/- 10.75 ml (30.2-380) P < 0.0001, representing a decrease of 72% of the volume infused. The nucleated cell count harvested was 1.6 x 10(10) +/- 0.1 (0.56-3.2), while the count infused was 6.9 x 10(9) +/- 0.1 (0.12-5.4) P < 0.0001. The median mononuclear cell count (MNC) per kg harvested was 0.67 x 10(8) +/- 0.05 (0.18-2.0) vs an infused cell number of 1.3 x 10(8) MNC/kg +/- 0.1 (0.6-33.6) P < 0. 0001. The CD34+ cells harvested were 2.8 x 10(6)/kg +/- 0.1 (0.25-10.2) vs an infused number of 6.0 x 10(6)/kg +/- 0.5 (0.84 31.0) P < 0.0001. The viability before and after processing was 99%. Red cell salvage performed in a semi-automated closed system is safe and reduces the risk of post-bone marrow harvest anemia in pediatric donors, decreases the volume infused into the donor and enriches the mononuclear and CD34+ cell population, without affecting hematopoietic reconstitution. PMID- 10467329 TI - Simultaneous cord blood transplantation of ex vivo expanded together with non expanded cells for high risk leukemia. AB - In the absence of a donor alternative a stem cell transplantation consisting of two cord blood components originating from the haploidentical brother was performed in a 2-year-old girl with c-ALL, early CNS relapse and 7% of blast cells in the BM 14 days before transplantation. Because of various ongoing infectious complications at that time, 1/8 of the immunogenetically acceptable sibling cord blood was ex vivo expanded 10 days before the transplantation date. The total CB consisting of 1.17 x 10(9) NC was cryopreserved in four separate bags. The one containing 1/8 of the total CB with 1.4 x 10(8) NC CliniMACS selected CD34+ cells was expanded in the presence of 100 ng/ml G-CSF, 100 ng/ml TPO and 100 ng/ml flt3-L in 10% autologous CB plasma and X-VIVO 10 medium at day 10 before transplantation. This expanded cell population was sterile and consisted of about 60% granulocytic cells (CD13+, CD15+), about 30% myelomonocytic cells (CD14, HLA-DR+), 5.2% megakaryocytes (CD61+) and 1.2% CD34+ cells. The proportion of T (CD3+), NK cells (CD56+) as well as dendritic cells (CD83+) was below 0.2%. The unseparated CB infused at day 0 and +1 consisted of a total of thawed 4.4 x 10(7) NC/kg BW, 5.8 x 10(4) CFU-GM/kg BW, 1.54 x 10(5) CD34+cells/kg BW and 7. 73 x 10(2) LTC-IC/kg BW. In addition, the 1 x 10(7) NC/kg BW ex vivo expanded cells representing 1.9 x 10(4) CFU-GM/kg BW, 1.13 x 10(5) CD34 cells/kg BW and 4.37 x 10(2) LTC-IC/kg BW, were infused at day +1. At day +2 after transplantation the patient revealed a focal pneumonia on X-ray with generalized sepsis and became catecholamine dependent. From day +4 the patient received 280 microg/m2 G-CSF. At day +5 she developed an erythroderma, which could not be identified as acute GVHD by biopsy. Early engraftment with leukocyte counts at days 8 and 14 were 350 and 700/microl, ANC 310 and 410/microl, respectively. Donor cells determined by chimerism analysis were 97% and 98% in the periphery at this early time. Most importantly, the pneumonia as well as the septicemia subsided within a few days. Notably, as well is the clearly shortened aplastic phase observed after this simultaneous CB cell component transplantation. The patients T cell and NK cell reconstitution could be detected at day +37 with 330 CD3+ cells/microl and 40 CD56+ cells/microl, respectively. The time to reach an absolute platelet count of 20 000 (50 000)/microl was 75 (103) days. The disease-free survival now exceeds 1 year in complete remission without chronic GVHD or any other health problems. These data show that the applicability of ex vivo expanded committed progenitors and LTC-IC, even in high risk leukemia at the time of transplantation, is feasible and can provide sufficient myeloid progenitors resulting in rapid engraftment able to clear bacterial pneumonia and sepsis. In addition, accelerated hematopoietic reconstitution apparently served as a well functioning platform for definitive graft-versus-leukemia activity. This transplantation of defined ex vivo generated components presents a feasible and generally applicable approach and may open a promising new avenue for cell therapy in malignant diseases. PMID- 10467328 TI - Allogeneic bone marrow transplant or second autograft in patients with acute leukemia who relapse after an autograft. Acute Leukaemia Working Party of the European Group for Blood and Marrow Transplantation (EBMT). AB - Among 2752 patients with acute leukemia who had recurrent leukemia after autograft in remission and were reported to the EBMT, 94 underwent an allogeneic bone marrow transplant and 74 received a second autograft. Recipients of HLA mismatched related or unrelated bone marrow had an increased transplant-related mortality (TRM, P = 0.017) and a decreased leukemia-free survival (LFS, P = 0.03), compared to recipients of HLA matched related or unrelated bone marrow. Outcome in recipients of HLA-compatible related or unrelated bone marrow was compared to those receiving a second autograft. TRM at 2 years was 51 +/- 8% in recipients of matched allografts and 26 +/- 6% following a 2nd autograft (P < 0.05). Two-year LFS was 27 +/- 7% and 35 +/- 6% in the two groups, respectively (NS). Multivariate analysis in these two groups showed that TRM was increased in patients who were in 2nd or later remission at 1st autograft (P < 0. 05) and allograft recipients (P < 0.05). Relapse was more common in patients with ALL (P < 0.001), above 25 years of age (P < 0.02), autograft performed later than 1991 (P < 0.05), and in second autografts (P < 0.05). LFS was decreased in patients >25 years of age (P < 0.01), if the interval from first autograft to relapse was 8 months or less (P < 0.01) and if TBI was used at first autograft (P < 0.05). PMID- 10467330 TI - Low incidence of molecular evidence for tumour in PBPC harvests from patients with high risk Ewing tumours. AB - Reverse transcriptase polymerase chain reaction (RT-PCR) was applied to evaluate the frequency of tumour cells in PBPC products from 15 high risk Ewing tumour (ET) patients who were treated according to EICESS 92 with high-dose chemotherapy (HDC) and stem cell rescue. Initial tumour cell contamination of the bone marrow (BM) detected by light microscopy was found in five and by RT-PCR in eight cases. RT-PCR was performed on each PBPC sample repeatedly at a sensitivity comparable to 20-100 highly EWS-Fli1 expressing tumour cells per 10 ml of fresh blood. Irrespective of the extent of BM involvement at diagnosis, all BM samples obtained before harvest were RT-PCR negative. Among 12 of 35 analysed apheresis products with single positive RT-PCR results only one sample tested reproducibly positive for tumour cell contamination in independent determinations. These preliminary data suggest that tumour cell contamination of PBPC is rarely found in patients with ET. PMID- 10467331 TI - Telomere length changes in patients undergoing hematopoietic stem cell transplantation. AB - Telomere length indicates the replicative history of cells, serving as a molecular measure of the replicative potential remaining in cells. To investigate telomere length changes in hematopoietic stem cells, patients undergoing hematopoietic stem cell transplantation (HSCT) were evaluated. Fifteen patients after allogeneic bone marrow transplantation (allo-BMT group), seven patients after autologous peripheral blood stem cell transplantation (auto-PBSCT group), and 39 healthy controls were studied. Telomere length was measured in peripheral mononuclear cells by Southern blot hybridization. There was no significant difference between the allo-BMT and the auto-PBSCT groups. In the allo-BMT group, the mean telomere length of recipients was 2.01 kb shorter than that of their donors (P = 0. 008), and was 1.59 kb shorter than that of age-matched putative normal controls (P = 0.002). Telomere shortening in the allo-BMT group was equivalent to 41.4 years of aging in the donors, and to 52. 4 years of aging in the normal controls. The mean telomere length in the auto-PBSCT group was 2.36 kb shorter than that of the age-matched putative controls (P = 0.043), which was equivalent to 61.5 years of aging in normal controls. The extent of telomere shortening in the allo-BMT group showed a trend to negative correlation with the number of mononuclear cells infused. These findings suggest that hematopoietic stem cells after HSCT lose telomere length and these shortened telomeres may result in a higher incidence of clonal disorders later in life. PMID- 10467332 TI - Combined anti-fungal therapy and surgical resection as treatment of pulmonary zygomycosis in allogeneic bone marrow transplantation. AB - Opportunistic fungal infection is a rare but severe complication in allogeneic bone marrow transplant (BMT) recipients. We report a 49-year-old patient who developed pneumonitis after BMT, due to a Mucorales fungus (class Zygomycetes), Absidia corymbifera. Infections due to mucormycosis are likely to become increasingly recognized even though the occurrence after BMT has only been described sporadically. We postulate that the patient was contaminated before BMT despite no intensive drug treatment or other iatrogenic features, related to his poor living conditions and developed the infection during aplasia. He immediately received i.v. liposomal amphotericin B (AmBisome) and GM-CSF. Because there was no response, the infected area and necrotic tissue were resected. Despite initial clinical and biological improvement and the absence of Mucor on mycological examination post-surgery, the patient died 3 weeks later from bilateral pulmonary infection and multiorgan failure. PMID- 10467333 TI - Ganciclovir-induced encephalopathy in a bone marrow transplant recipient. AB - Ganciclovir is widely used as prophylactic and pre-emptive therapy, as well as treatment, for CMV infection following BMT. We report a case treated with ganciclovir 5 days a week. Following escalation of the ganciclovir dose to a twice daily dose to treat CMV antigenaemia, he developed encephalopathy. His encephalopathy resolved with withdrawal of ganciclovir. Ganciclovir encephalopathy has been described in other groups of patients but has not been reported following BMT to date. With its widespread use this complication is likely to be seen more often. PMID- 10467334 TI - A case of hemophagocytic lymphohistiocytosis with prolonged remission after syngeneic bone marrow transplantation. AB - We report a 7-year-old girl with hemophagocytic lymphohistiocytosis who received a syngeneic bone marrow transplant from her twin sister. She presented with high fever and cough. Laboratory findings revealed pancytopenia, elevation of liver enzymes, and hyperferritinemia. Bone marrow examination revealed histiocytic hemophagocytes and lymphoblastoid cells. Southern blot analysis of the bone marrow cells revealed a monoclonal proliferation of EBV-infected lymphocytes. Although she underwent combined chemotherapy according to the HLH-94 protocol, she developed severe pancytopenia. Following myeloablative conditioning with busulfan (16 mg/kg), cyclophosphamide (120 mg/kg), and etoposide (1.5 g/m2), she was transplanted with 6.6 x 10(8)/kg mononuclear cells from the twin sister. She remains in complete remission 23 months after transplantation. PMID- 10467335 TI - Ultrastructural changes in peripheral blood neutrophils in a patient receiving ganciclovir for CMV pneumonitis following allogenic bone marrow transplantation. AB - A 13-year-old splenectomized, multitransfused beta-thalassemia major, male patient received an allogenic BMT from his HLA-compatible brother after suffering grade III regimen-related pulmonary toxicity. He developed features of CMV pneumonitis with positive pp65 CMV antigenemia involving 2.5% peripheral blood neutrophils from day +46. The patient received intravenous immunoglobulin and ganciclovir 5 mg/kg intravenously twice daily. His neutrophil count was maintained above 1 x 10(9)/l by G-CSF 5 microg/kg subcutaneously as and when required. From day 7 onwards following twice daily ganciclovir his peripheral blood smear started showing isolated cytoplasmic inclusions, 1-3 per neutrophil, 3-5 mu in diameter, involving 2-3% of the neutrophils and occasional monocytes. Transmission election microscopy of peripheral blood neutrophils showed type I and type II intranuclear inclusions. These inclusions disappeared within 48 h of stopping ganciclovir. Inclusions were not seen in three patients who were given prophylactic ganciclovir 5 mg/kg once daily for 5 days every week following allogenic BMT after the same conditioning regimen. These patients were also negative for CMV antigenemia. Development of type I and type II intranuclear inclusions in blood neutrophils in patients receiving ganciclovir therapy has not been reported previously, and the striking light microscopic changes provide simple morphological evidence of the toxic effect of this drug on blood neutrophils. PMID- 10467336 TI - Improvement of amyloid-related symptoms after autologous stem cell transplantation in a patient with hepatomegaly, macroglossia and purpura. AB - AL amyloidosis was diagnosed in a 56-year-old woman with spontaneous purpura, macroglossia and hepatomegaly, a serum IgGk monoclonal gammopathy and a 25% plasma cell bone marrow infiltration. She was started on high-dose treatment consisting of four monthly cycles of VID chemotherapy, then underwent a stem cell collection after priming with cyclophosphamide + G-CSF. Myeloablative therapy was with melphalan and busulfan. Hematologic recovery was fast and uncomplicated. At follow-up 22 months from ASCT, the patient shows a complete remission of the clonal plasma cell disorder, normalization of liver size and alkaline phosphatase level and a significant improvement in the signs of vascular and soft tissue amyloid infiltration. PMID- 10467337 TI - Successful related umbilical cord blood transplantation for graft failure following T cell-depleted non-identical bone marrow transplantation in a child with major histocompatibility complex class II deficiency. AB - Major histocompatibility complex (MHC) class II deficiency is a rare form of primary combined immunodeficiency that can only be corrected by stem cell transplantation. We report a 4(1/2)-year-old girl with MHC class II deficiency who underwent a related CBT due to graft failure following T cell-depleted non identical BMT. The patient is alive and well 2 years after the second transplant. A sustained hematopoietic engraftment and a progressive immune recovery have been detected. We conclude that cord blood may be an effective source of hematopoietic stem cells for patients with immuno- deficiency disorders including diseases with a high rate of graft failure. PMID- 10467338 TI - Flow cytometry comparison of CD34+ subsets in bone marrow and peripheral blood after priming with glycosylated or non-glycosylated rhG-CSF. PMID- 10467339 TI - Response to the comment on 'Flow cytometry comparison of CD34+ subsets in bone marrow and peripheral blood after priming with glycosylated or non-glycosylated rhG-CSF' by JM miclea PMID- 10467340 TI - Post-transplant Epstein-Barr virus-associated meningoencephalitis and lymphoid interstitial pneumonitis. PMID- 10467341 TI - Allogeneic peripheral blood stem cell transplantation in a Wiskott-Aldrich syndrome patient. PMID- 10467342 TI - Multifocal reversible epithelial dysplasia mimicking carcinoma in situ after conditioning therapy with busulfan and cyclophosphamide. PMID- 10467343 TI - Poor stem cell mobilizers--variation in response to G-CSF dose escalation. PMID- 10467344 TI - Galectins: an evolutionarily conserved family of animal lectins with multifunctional properties; a trip from the gene to clinical therapy. AB - Galectins constitute a family of evolutionarily conserved animal lectins, which are defined by their affinity for poly-N-acetyllactosamine-enriched glycoconjugates and sequence similarities in the carbohydrate recognition domain. During the past decade, attempts to dissect the functional role for galectins in vivo have been unsuccessful in comparison to the overwhelming information reached at the biochemical and molecular levels. The present review deals with the latest advances in galectin research and is aimed at validating the functional significance of these carbohydrate-binding proteins. Novel implications of galectins in cell adhesion, cell growth regulation, immunomodulation, apoptosis, inflammation, embryogenesis, metastasis and pre-mRNA splicing will be particularly discussed in a trip from the gene to the clinical therapy. Elucidation of the molecular mechanisms involved in galectin functions will certainly open new avenues not only in biomedical research, but also at the level of disease diagnosis and clinical intervention, attempting to delineate new therapeutic strategies in autoimmune diseases, inflammatory processes, allergic reactions and tumor spreading. PMID- 10467345 TI - Novel aspects of Ras proteins biology: regulation and implications. AB - The importance of Ras proteins as crucial crossroads in cellular signaling pathways has been well established. In spite of the elucidation of the mechanism of RAS activation by growth factors and the delineation of MAP kinase cascades, the overall framework of Ras interactions is far from being complete. Novel regulators of Ras GDP/GTP exchange have been identified that may mediate the activation of Ras in response to changes in intracellular calcium and diacylglycerol. The direct activation of Ras by free radicals such as nitric oxide also suggests potential regulation of Ras function by the cellular redox state. In addition, the array of Ras effectors continues to expand, uncovering links between Ras and other cellular signaling pathways. Ras is emerging as a dual regulator of cellular functions, playing either positive or negative roles in the regulation of proliferation and apoptosis. The signals transmitted by Ras may be modulated by other pathways triggered in parallel, resulting in the final order for proliferation or apoptosis. The diversity of ras-mediated effects may be related in part to differential involvement of Ras homologues in distinct cellular processes. The study of Ras posttranslational modifications has yielded a broad battery of inhibitors that have been envisaged as anti-cancer agents. Although an irreversible modification, Ras isoprenylation appears to be modulated by growth factors and by the activity of the isoprenoid biosynthetic pathway, which may lead to changes in Ras activity. PMID- 10467346 TI - Butyrate mediates Caco-2 cell apoptosis via up-regulation of pro-apoptotic BAK and inducing caspase-3 mediated cleavage of poly-(ADP-ribose) polymerase (PARP). AB - Butyrate exerts potent anti-tumor effects by inhibiting cancer cell growth and inducing apoptosis. However, the molecular mechanisms mediating these effects remain largely unknown. Using the Caco-2 cell line, a well established model of colon cancer cells, our data show that butyrate induced apoptosis (maximum 79%) is mediated via activation of the caspase-cascade. A key event was the proteolytic activation of caspase-3, triggering degradation of poly-(ADP-ribose) polymerase (PARP). Inactivation of caspase-3 with the tetrapeptide zDEVD-FMK completely inhibited the apoptotic response to butyrate. In parallel, butyrate potently up-regulated the expression of the pro-apoptotic protein bak, without changing Caco-2 cell bcl-2 expression. Butyrate-induced Caco-2 cell apoptosis was completely blocked by the addition of cycloheximide, indicating the necessity of protein synthesis. However, when this inhibitor was added at a time point where bak expression was already enhanced (12 - 16 h after butyrate stimulation), it failed to protect Caco-2 cells against apoptosis. Taken together, these data provide evidence that the molecular events involved in butyrate induced colon cancer cell apoptosis include the caspase-cascade and the mitochondrial bcl pathway. PMID- 10467347 TI - Benzamide riboside induces apoptosis independent of Cdc25A expression in human ovarian carcinoma N.1 cells. AB - One of the mechanisms of action of a new oncolytic agent, benzamide riboside (BR) is by inhibiting inosine 5'-monophosphate dehydrogenase (IMPDH) which catalyzes the formation of xanthine 5'-monophosphate from inosine 5'-monophosphate and nicotinamide adenine dinucleotide, thereby restricting the biosynthesis of guanylates. In the present study BR (10 - 20 microM) induced apoptosis in a human ovarian carcinoma N.1 cell line (a monoclonal derivative of its heterogenous parent line HOC-7). This was ascertained by DNA fragmentation, TUNEL assay, [poly(ADP)ribose polymerase]-cleavage and alteration in cell morphology. Apoptosis was accompanied by sustained c-Myc expression, concurrent down regulation of cdc25A mRNA and protein, and by inhibition of Cdk2 activity. Both Cdk2 and cdc25A are G1 phase specific genes and Cdk2 is the target of Cdc25A. These studies demonstrate that BR exhibits dual mechanisms of action, first by inhibiting IMPDH, and second by inducing apoptosis, which is associated with repression of components of the cell cycle that are downstream of constitutive c Myc expression. PMID- 10467348 TI - Structure and promoter analysis of murine CAD and ICAD genes. AB - Caspase-activated DNase (CAD) degrades chromosomal DNA during apoptosis, whereas ICAD (inhibitor of CAD) inhibits the CAD's DNase by binding to it. Here, we describe the assignment of murine CAD and ICAD genes to the distal part of murine chromosome 4. Molecular cloning and structural analysis indicated that CAD and ICAD genes are comprised of 7 and 6 exons, respectively. Two different ICAD mRNAs coding for two forms of ICAD proteins (ICAD-S and ICAD-L) were found to be produced by alternative splicing of intron 5. The CAD and ICAD mRNAs were detected ubiquitously in various murine tissues. Analyses of the promoter activity with a series of deletion mutants of their 5' flanking regions indicated that a 190-bp 5' flanking region of the CAD gene was sufficient to promote the transcription. Whereas, a 120-bp flanking region of ICAD gene was required to promote its transcription. These regions do not show similarity between CAD and ICAD genes, suggesting that expression of CAD and ICAD genes is regulated by different mechanisms. PMID- 10467349 TI - Overexpression of atypical PKC in PC12 cells enhances NGF-responsiveness and survival through an NF-kappaB dependent pathway. AB - Removal of atypical PKC blocks NGF-induced differentiation of PC12 cells.1 We now examine the consequences that overexpression of atypical PKCs had upon NGF responses. PC12 cells were stably transfected with either PKC-iota or PKC-zeta. Overexpression of atypical PKCs markedly enhanced NGF- induced neurite outgrowth as well as enhanced NGF-stimulated JNK kinase. Cotransfection of HA-JNK1 along with increasing concentrations of PKC-iota, resulted in dose-dependent phosphorylation of GST c-Jun (1 - 79). NGF treatment of PC12 cells resulted in activation of NF-kappaB. In comparison, overexpression of atypical PKC-iota was by itself sufficient to activate NF-kappaB and shift the kinetics of NGF-induced kappaB activity. Furthermore, transfection of full-length antisense PKC-iota blocked basal and NGF-stimulated NF-kappaB. Differentiated and undifferentiated PC12 cells overexpressing atypical PKC-iota were protected from serum deprivation induced cell death. Collectively, these findings demonstrate that atypical PKC iota lies in a pathway that regulates NF-kappaB and contributes to both neurotrophin-mediated differentiation and survival signaling. PMID- 10467350 TI - Overexpression of the p21 sdi1 gene induces senescence-like state in human cancer cells: implication for senescence-directed molecular therapy for cancer. AB - Normal cells in a culture enter a nondividing state after a finite number of population doubling, which is termed replicative senescence, whereas cancer cells have unlimited proliferative potential and are thought to exhibit an immmortal phenotype by escaping from senescence. The p21 gene (also known as sdi1), which encodes the cyclin-dependent kinase inhibitor, is expressed at high levels in senescent cells and contributes to the growth arrest. To examine if the p21sdi1 gene transfer could induce senescence in human cancer cells, we utilized an adenoviral vector-based expression system and four human cancer cell lines differing in their p53 status. Transient overexpression of p21sdi1 on cancer cells induced quiescence by arresting the cell cycle at the G1 phase and exhibited morphological changes, such as enlarged nuclei as well as a flattened cellular shape, specific to the senescence phenotype. We also showed that p21sdi1 transduced cancer cells expressed beta-galactosidase activity at pH 6.0, which is known to be a marker of senescence. Moreover, the polymerase chain reaction-based assay demonstrated that levels of telomerase activity were significantly lower in p21sdi1-expressing cells compared to parental cancer cells. These observations provide the evidence that p21sdi1 overexpression could induce a senescence-like state and reduce telomerase activity in human cancer cells, suggesting that these novel p21sdi1 functions may have important implications for anticancer therapy. PMID- 10467351 TI - Interferon-alpha induces apoptosis in human KB cells through a stress-dependent mitogen activated protein kinase pathway that is antagonized by epidermal growth factor. AB - We have demonstrated that interferon-alpha2-recombinant (IFNalpha) at growth inhibitory concentrations enhances the expression and signalling activity of the epidermal growth factor receptor (EGF-R) in human epidermoid carcinoma KB cells. Here we report that KB cells exposed to IFNalpha underwent apoptotic cell death and this effect was antagonized by EGF. We have also found that IFNalpha enhanced the expression of heat shock proteins (HSP) HSP-70, HSP-90 and HSP-27 and activated the NH2-terminal Jun kinase-1 (JNK-1) and p38 mitogen activated protein kinase, the target enzymes of a stress-dependent intracellular transduction pathway. Moreover, the overexpression of the wild-type JNK-1, obtained through plasmid transfection of KB cells, induced apoptosis which was potentiated by the exposure of wild-type JNK-1 (JNK-1wt)-transfected cells to IFNalpha. All these effects were neutralized by the addition of EGF to parental and JNK-1wt transfected KB cells exposed to IFNalpha. In conclusion, EGF has a protective effect on KB cells from apoptosis while antagonizing a stress response elicited by IFNalpha and targeted on the stress pathway terminal kinases. PMID- 10467352 TI - Bezafibrate as differentiating factor of human myeloid leukemia cells. AB - Bezafibrate belongs to the class of fibric acid derivatives usually used as antihyperlipidemia agents. From the biochemical point of view, these drugs show intriguing properties which leads one to think they may promote a differentiation process in tumour cells. This new pharmacological activity of fibrates could partially depend on the induction of an oxidative stress. To test this hypothesis, the effect of bezafibrate, as well as of clofibric acid and gemfibrozil, on growth, functional and cytochemical characteristics of human leukaemia-derived cell lines HL-60, U-937 and K-562 has been studied in some details. The results show that bezafibrate, gemfibrozil and clofibric acid, do induce differentiation in human myeloid leukaemia cell lines as indicated by several differentiation markers. Moreover fibrates, in dose dependent manner, significantly alter the cell cycle distributions, mainly leading to G0/G1 phase increment and G2/M phase reduction. The differentiating activity of fibrates could have significant implications both for the pharmacotoxicological profile of this class of compounds and for the pathophysiology of neoplastic disease. PMID- 10467353 TI - Ceramide-induced apoptosis occurs independently of caspases and is decreased by leupeptin. PMID- 10467354 TI - Characterization of Apt- cell lines exhibiting cross-resistance to glucocorticoid and Fas-mediated apoptosis. AB - Apoptosis induction by staurosporine, ceramide, and Fas stimulation was investigated in the mouse thymoma cell line W7.2 and a panel of dexamethasone (dex)-resistant W7.2 mutant cell lines, Apt3.8, Apt4.8 and Apt5.8, and a Bcl-2 transfected W7.2 cell line (Wbcl2). While W7. 2 cells were found to be sensitive to these apoptosis inducers, the Apt- mutants and Wbcl2 cells were shown to be resistant to some or all of the treatments. Specifically, all three Apt- mutants and Wbcl2 cells were found to be resistant to ceramide and Fas-mediated apoptosis, whereas, Apt4.8 and Apt5.8 were sensitive to staurosporine-induced apoptosis under conditions in which Apt3.8 and Wbcl2 cells were resistant. Measurements of caspase activity and cytochrome c release in cytosolic extracts of dex and staurosporine-treated cells indicated that the recessive Apt- mutations effect steps upstream of mitochondrial dysfunction. Steady-state RNA levels of apoptosis-associated gene transcripts showed that the observed differential resistance of the Apt- cell lines could not be explained by altered expression of numerous Bcl-2 or Fas related genes. Transient transfection of human Fas gene coding sequences into the Apt- mutants and Wbcl2 cells did not induce apoptosis, even though these same cell lines were sensitive to ectopic expression of the FADD and caspase 8 genes. Taken together, these data provide genetic evidence for the existence of shared components in the dex- and Fas mediated apoptotic pathways in W7.2 cells. PMID- 10467355 TI - Bacterial death induced by expression of the intracellular portion of human Fas. AB - In attempting to produce the intracellular portion of human Fas (IC175 - 319) as a GST-fusion protein we found that expression of GST-IC175 - 319, but not GST alone or GST-IC231 - 298 (containing the Fas death domain), rapidly caused the death of host E. coli cells. Expression of GST-IC175 - 319 with a single amino acid substitution (V238N) corresponding to the mouse lprcg mutation, or E245A, which abolishes the ability of Fas to self-associate, did not kill bacteria. Deletional analysis identified a 20-amino acids region (Asp210 - Lys230) as essential for the killing activity, and introduction of a single amino acid substitution (T225P) in this 20 amino acid region markedly decreased the ability of Fas- IC175 - 319 to cause bacterial death. These data indicate that Fas can deliver a death signal in prokaryotic organisms by a means that shares some features with eukaryotic cells, and raise the possibility that certain mechanisms leading to programmed cell death may be conserved from bacteria to mammalian cells. PMID- 10467356 TI - IL-4 inhibits apoptosis and prevents mitochondrial damage without inducing the switch to necrosis observed with caspase inhibitors. AB - We previously demonstrated that the broad-spectrum caspase inhibitor, zVAD-fmk, totally deviated apoptosis to necrosis in B lymphocytes. We report here that, in contrast with zVAD-fmk, IL-4 protected B cells from spontaneous and from dexamethasone-induced apoptosis and actually maintained cell viability. This was assessed by morphological and biochemical criteria and accompanied by the maintenance of mitochondrial transmembrane potential (DeltaPsiCm) and elevated glutathione (GSH) levels. Under these conditions, zVAD-fmk also totally inhibited apoptosis in thymocytes, but it partly preserved cell viability with a parallel increase in the percentage of cells exhibiting high DeltaPsiCm and elevated GSH levels. Nevertheless, non-rescued cells were deviated to necrosis. Therefore, the pathway leading to either apoptosis or necrosis appears to involve common mitochondrial dysfunctions which could not be reversed by caspase inhibition, suggesting that the pharmacological inhibition of cell death should occur at an earlier stage. PMID- 10467357 TI - Opportunities and challenges in targeting genes for therapy. PMID- 10467358 TI - Adenoviral vectors: prospects for gene delivery to the central nervous system. PMID- 10467359 TI - A model for long-term transgene expression in spinal cord regeneration studies. AB - In order to determine the suitability of first generation adenoviral vectors for gene delivery into spinal cord white matter, four different titres of beta galactosidase-expressing adenovirus were injected into spinal cord white matter of adult rats. At titres > or =106 p.f.u., transgene expression was extensive but severe tissue damage was observed in the form of axon degeneration, demyelination and astrocyte loss. When < or = 105 p.f.u. were injected, only low levels of axon degeneration and demyelination were observed. beta-Galactosidase activity was detectable at 72 days and did not diminish significantly with time. The immune response in the spinal cord to 105 p.f.u. over 72 days was minimal, and indistinguishable from that to injection of buffer. A prominent immune response was observed when 107 p.f.u. was injected into the spinal cord of PVG rats, and when 105 or 107 p.f.u. was injected into AO rats. These results indicate that the immune response in PVG rats to betagal-expressing adenovirus is both strain and titre dependent. First generation adenoviral vectors, therefore, induce moderate and long-lived transgene expression with minimal tissue damage and immune response when an appropriate titre is injected into the low responder PVG rat strain, providing a suitable model for assessing the effect of gene delivery in models of spinal cord injury. PMID- 10467360 TI - Adenoviral vectors for in vivo gene delivery to oligodendrocytes: transgene expression and cytopathic consequences. AB - Replication defective viral vectors provide a potentially useful means of gene transfer to oligodendrocytes and thus for studying the pathogenesis of white matter disease. In this study we have examined the expression pattern of E1/E3 deleted adenoviral vectors expressing the reporter gene LacZ (AdlacZ) as a means of establishing the value of these vectors for gene delivery to oligodendrocytes in adult rat white matter. Our results indicate that although such an approach can be used to induce transgene expression in oligodendrocytes, it is complicated by both immunogenic and cytopathic effects. Thus, in normal animals, injection of DeltaE1/E3 adenoviral vectors was associated with a robust immune response that led to a lack of expression by 40 days after injection. In order to overcome this complication, virus was injected into the white matter of immuno-deficient athymic rats. These experiments indi- cated that even in the absence of a T cell response high viral titres of DeltaE1/E3 adenoviral vectors had a profound cytopathic effect leading to death of oligodendrocytes and hence demyelination. A similar cytopathic effect was demonstrated using an adenoviral vector expressing the neurocytokine ciliary neurotrophic factor (AdCNTF). As the titre of injected virus was decreased there was a significant decrease in the number of transgene expressing cells. These experiments therefore indicated that in immunodeficient recipients there is a narrow window of virus titre that results in a high rate of infectivity and expression without significant cytopathic consequences. At higher viral titres the cytopathic effects include oligodendrocyte death and demyelination, while at lower titres there is a significant decrease in the efficiency of the number of cells expressing the transgene. PMID- 10467361 TI - Inflammatory responses and their impact on beta-galactosidase transgene expression following adenovirus vector delivery to the primate caudate nucleus. AB - An E1, E3 deleted adenovirus vector, serotype 5, carrying the marker gene LacZ was bilaterally microinfused into the caudate nuclei of 10 St Kitts green monkeys. The location and number of cells expressing transgene and host immunologic response were evaluated at 1 week (n = 2) and 1 month (n = 8) following vector infusion. A large number of cells expressed beta-galactosidase in some monkeys, exceeding 600000 in one monkey, but no expression was seen in three of 10. All monkeys had positive adenoviral antibody titers before vector infusion, indicating the possibility of previous exposure to some adenovirus, but only one showed a significant increase in titer afterwards. Inflammatory cell markers revealed an inverse correlation between transgene expression and the extent of inflammatory response. Dexamethasone administered immediately before and for 8 days following vector delivery, however, had no effect on transgene expression. The demonstration of significant inflammatory responses in the brain of some individual primates, including demyelination, indicates the need for new generations of adenovirus vectors, or the successful suppression of inflammatory responses, before this vector is suitable for non-cytotoxic clinical applications in the CNS. PMID- 10467362 TI - Improved packing of poly(ethylenimine)/DNA complexes increases transfection efficiency. AB - We have developed a modified poly(ethylenimine) (PEI) transfection procedure that significantly increases PEI's transfection efficiency. While the basic transfection procedure had a transfection efficiency of 37%, our modified procedure yielded a 53% transfection efficiency. The altered procedure gives improved results because of two simultaneous actions: free polycations are removed from the transfecting solutions, and the composition of the PEI complexes that are administered to cells has been modified. The reduction in the amount of free polycations in transfecting solutions reduced the toxicity sometimes associated with the administration of polycations to cellular environments. The structural modification of PEI/DNA transfecting complexes involves improved PEI packing around the delivered plasmid to yield a greater buffering capacity without a change in the complex's surface charge concentration. These structural properties were confirmed by titration and zeta potential analyses. Whether the modified PEI/DNA complexes are more effective because of increased cellular uptake or an enhanced ability to escape from endolysosomes has been addressed. The increase in transfection efficiency was obtained when the buffering capacity of the PEI/DNA was increased without a change in surface charge concentration, which implies that it is the property of enhanced lysosomal buffering that is responsible for successful PEI transfection. PMID- 10467363 TI - Distribution of retroviral vectors and vector producer cells using two routes of administration in rats. AB - The clinical use of retroviral vector producer cells (VPCs) to deliver retroviral vectors efficiently to target cells has been investigated as a method to increase efficiency of gene delivery, presumably as a result of continued vector production in vivo. Studies were conducted in rats to evaluate the distribution of vector to distal organs and tissues as measured by transduction. Rats were treated with two doses of VPCs using two routes of administration: (1) subcutaneous injection, chosen to maximize both the dose and exposure of animals, thereby enabling identification of potential target organs under worst-case conditions; and (2) direct injection into brain parenchyma, chosen to mimic the intended clinical route of administration and provide an estimate of risk to patients receiving this therapy. Twelve organs or tissues were collected 7 days after administration of VPCs and analyzed by PCR for the presence of vector and vector producer cell sequences. Vector was detected most frequently at the site of injection by either route of administration. Less frequently, vector was detected in draining lymph nodes at the higher dose only using either route of injection. Single specimens of lung and contralateral skin were positive for vector following subcutaneous administration only. Vector was detected in gonadal tissue from a single low-dose male following subcutaneous administration, but this finding was not reproduced in any high-dose male or any males injected intracerebrally. In contrast, VPCs were detected only at the site of administration. The frequency of detection of VPCs 7 days after administration was higher when rats were injected by the intracerebral route. Based on these studies, gene transfer to distal organs or gonadal tissue following intracerebral administration of VPCs is not considered to be a risk to patients undergoing retroviral vector gene therapy for the treatment of brain cancer (glioblastoma multiforme; GBM). PMID- 10467364 TI - Human serum-resistant retroviral vector particles from galactosyl (alpha1-3) galactosyl containing nonprimate cell lines. AB - Retroviral vector particles (RVP) which are resistant to inactivation by human serum will be needed for many in vivo gene therapy applications. Murine-based producer cell lines generate RVP which are inactivated by human serum, reportedly due to the presence of the galactosyl (alpha1-3) galactosyl carbohydrate moiety (alphaGal) on these and other nonprimate producer cells and RVP. Consequently, human cells (which lack the alphaGal moiety) have been developed as producer cell lines for generation of human serum-resistant RVP. In this study, we report that contrary to earlier reports, the presence of the alphaGal moiety on producer cells and RVP does not necessarily correlate with cell killing or RVP inactivation by human serum. We show that the alphaGal-positive ferret brain cell line, Mpf, is an excellent basal cell line for generation of RVP which have titers and serum resistance levels equal to or greater than RVP produced in human cell lines such as HT1080. Therefore, packaging cell lines need not be limited to those of human or primate origin for production of human serum-resistant RVP. PMID- 10467365 TI - Local and distant transfection of mdx muscle fibers with dystrophin and LacZ genes delivered in vivo by synthetic microspheres. AB - Patterns of dystrophin and beta-galactosidase expression were examined in mdx mice after i.m. injections of synthetic microspheres (MF-2) loaded with full length (pHSADy) or mini-dystrophin gene (pSG5dys) cDNA plasmid constructs or with LacZ marker gene (pCMV-LacZ). A single injection of 25 microg pHSADy into quadriceps femoris muscle resulted in 6.8% of dystrophin positive myofibers (DPM) in a given muscle; 8.4% of DPM in glutaeus muscle and 4.3% of DPM in quadriceps femoris muscle of contralateral limb on day 21 after exposure compared with only 0.6% DPM in intact (non-injected) mdx mice. A high proportion of DPM (17.6% and 10.8%, respectively) was registered in both injected and contralateral muscles after mini- gene cDNA administration. MF-2/dystrophin cDNA particles were detected by FISH analysis in about 60-70% of myofiber nuclei in muscles of injected and contralateral limbs 7 days after application. The presence of human dystrophin cDNA and its products in all skeletal muscles and in different internal organs was proven by PCR and RT-PCR analysis. Patches of beta galactosidase expression were abundant in injected muscle, and frequent in the contralateral and other skeletal muscles as well as in diaphragm, heart and lungs. High levels of dystrophin cDNA expression, and an efficient distant transfection effect with preferential intranuclei inclusion of MF-2 vehicle, are very encouraging for the development of a new constructive strategy in gene therapy trials of DMD. PMID- 10467366 TI - Enhancement of tumor ablation by a selected HSV-1 thymidine kinase mutant. AB - With the advent of gene therapy, herpes simplex virus type I (HSV-1) thymidine kinase (TK) has garnered much interest as a suicide gene for cancer ablation. As a means to improve the overall efficacy of the prodrug-gene activation approach, as well as to reduce ganciclovir-mediated toxicity, a large library of mutant thymidine kinases was generated and screened for the ability to enhance in vitro cell sensitivity to the prodrugs, ganciclovir (GCV) and acyclovir (ACV). Enzyme kinetics of one thymidine kinase mutant from this library that contains six amino acid substitutions at or near the active site reveals a distinct mechanism for providing enhanced prodrug-mediated killing in mammalian cells. In in vitro rat C6 cell prodrug sensitivity assays the TK mutant (mutant 30) achieves nanomolar IC50 values with GCV and ACV, in contrast to IC50values of 30 microM and >100 microM, respectively, for wild-type TK. In a mouse xenograft tumor model, growth of mutant 30 expressing tumors is restricted by ganciclovir at a dose at least 10 fold lower than one that impedes growth of wild-type TK-expressing tumors. Furthermore, in the presence of GCV a substantial bystander effect is observable when only 20% of the tumor cells express mutant 30 whereas no restriction in tumor growth is seen in tumors bearing the wild-type TK under the same conditions. The enhanced sensitization to prodrugs conferred by mutant 30 is apparently due to a 35-fold increase in thymidine Km which results in reduced competition between prodrug and thymidine at the active site. This provides mutant 30 a substantial kinetic advantage despite very high Kms for both ganciclovir and acyclovir. Molecular modeling of the mutations within the active site suggests that a tyrosine substitution at alanine 168 (A168) alters thymidine and prodrug interactions by causing catalytically important residues to move. The use of mutant 30 in place of the wild-type TK should provide a more effective gene therapy of cancer. PMID- 10467367 TI - Cellular redox state alters recombinant adeno-associated virus transduction through tyrosine phosphatase pathways. AB - Several types of environmental damage including UV, hydroxyurea and ionizing irradiation have been shown to augment rAAV transduction. Current hypotheses suggest that these environmental stimuli lead to the enhanced production and/or activation of cellular factors important in the conversion of single-stranded DNA genomes to expressible forms. However, the mechanisms of action are currently unknown. We hypothesized that reactive oxygen intermediates (ROI) may play a common role in the augmentation of rAAV transduction by these environmental stimuli. Our results demonstrate that treatment with hydrogen peroxide can give equivalent or greater levels of augmentation in rAAV transduction as that seen by hydroxyurea or UV irradiation. For all environmental stimuli, pretreatment with the hydroxyl radical (H0 small middle dot) scavenger, N-acetyl-L-cysteine (NAC), completely blocked augmentation of rAAV transduction. Furthermore, using electron spin resonance spectroscopy (ESR), we demonstrated that both UV and H2O2 treatment of cell lines lead to the induction of H0 small middle dot radicals. Our results demonstrating that NaOV inhibits the augmentation of rAAV transduction following UV and H2O2 treatment, implicate H0 small middle dot radicals as modulators of tyrosine phosphatase pathways involved in rAAV transduction. Alterations in the cellular redox state and subsequent activation of tyrosine phosphatase pathways appear to alter the phosphorylation status of the previously identified single-stranded sequence binding protein (ssD-BP), with reduced phosphorylation correlating with an enhancement in rAAV transduction. In summary, we conclude that the cellular redox state may play an important role in regulating rAAV transduction. PMID- 10467368 TI - Stabilized plasmid-lipid particles for regional gene therapy: formulation and transfection properties. AB - Previous work (Wheeler et al, Gene Therapy 1999; 6: 271-281) has shown that plasmid DNA can be entrapped in 'stabilized plasmid-lipid particles' (SPLP) containing the fusogenic lipid dioleoylphosphatidylethanolamine (DOPE), low levels (5-10 mol%) of cationic lipid, and stabilized by a polyethyleneglycol (PEG) coating. The PEG moieties are attached to a ceramide anchor containing an arachidoyl acyl group (PEG-CerC20). These SPLP exhibit low transfection potencies in vitro, due in part to the long residence time of the PEG-CerC20 on the SPLP surface. In this work we employed SPLP stabilized by PEG attached to ceramide containing an octanoyl acyl group (PEG-CerC8), which is able to quickly exchange out of the SPLP, to develop systems that give rise to optimized in vitro and in vivo (regional) transfection. A particular objective was to achieve cationic lipid contents that give rise to maximum transfection levels. It is shown that by performing the dialysis procedure in the presence of increasing concentrations of citrate, SPLP containing up to 30 mol% of the cationic lipid dioleoydimethylammonium chloride (DODAC) could be generated. The SPLP produced could be isolated from empty vesicles by sucrose density gradient centrifugation, and exhibited a narrow size distribution (62 +/- 8 nm, as determined by freeze fracture electron microscopy) and a high plasmid-to-lipid ratio of 65 microg/micromol (corresponding to one plasmid per particle) regardless of the DODAC content. It was found that isolated SPLP containing 20-24 mol% DODAC resulted in optimum transfection of COS-7 and HepG2 cells in vitro, with luciferase expression levels comparable to those achieved for plasmid DNA cationic lipid complexes. In vivo studies employing an intraperitoneal B16 tumor model and intraperitoneal administration of SPLP also demonstrated maximum luciferase expression for DODAC contents of 20-24 mol% and significantly improved gene expression in tumor tissue as compared with complexes. We conclude that SPLP stabilized by PEG-CerC8 and containing 20-24 mol% cationic lipid are attractive alternatives to plasmid DNA-cationic lipid complexes for regional gene therapy applications. PMID- 10467369 TI - The effect of CpG sequences on the B cell response to a viral glycoprotein encoded by a plasmid vector. AB - The effect of palindromic CpG sequences on the B cell response to plasmid vectors expressing a highly immunogenic viral glycoprotein was investigated. Methylation of the CpG sequences of bacterial expression vectors abolished their ability to induce an antibody response to the transgene product in mice. The antibody response could be rescued by concomitant injection of oligonucleotides carrying immunostimulatory sequences. The B cell response to two plasmid vectors, both expressing the same viral glycoprotein but containing a different content of the highly stimulatory AACGTT motif, was compared. Comparable B cell responses were induced to the two constructs given at an optimal vaccine dose while the vector containing additional palindromic sequences resulted in higher antibody titers at a suboptimal dose. These data indicate that deletion of CpG motifs or methylation of such sequences in plasmid DNA can abrogate the immune response to the vector encoded antigen and might thus enhance their usefulness as gene therapy vehicles. PMID- 10467370 TI - Adenovirus-mediated expresssion of the murine ecotropic receptor facilitates transduction of human hematopoietic cells with an ecotropic retroviral vector. AB - One factor limiting the ability to modify human repopulating hematopoietic cells genetically with retroviral vectors is the relatively low expression of the cognate viral receptor. We have tested sequential transduction of human hematopoietic cells with an adenoviral vector encoding the ecotropic retroviral receptor followed by transduction with an ecotropic retroviral vector. Adenoviral transduction of K562 erythroleukemia cells was highly efficiently with >95% of cells expressing the ecotropic receptor at a multiplicity of infection (MOI) of 103with a correspondingly high transduction with a retroviral vector. Ecotropic receptor expression in CD34+ cells following transduction with adenoviral vectors was increased by at least two-fold (from 20 to 48%) by replacing the RSV promoter with the CMV E1a promoter, resulting in a parallel increase in retroviral transduction efficiency. Replacing the head portion of the fiber protein in conventional adenoviral vectors (serotype 5) with the corresponding portion from an adenoviral 3 serotype resulted in ecotropic receptor expression in 60% of CD34+ cells at an MOI of 104 and a retroviral transduction of 60% of hematopoietic clonogenic progenitors. The sequential transduction strategy also resulted in efficient transduction of the primitive CD34+CD38- subset suggesting that it may hold promise for genetic modification of human hematopoietic stem cells. PMID- 10467371 TI - Tumor-specific gene transfer via an adenoviral vector targeted to the pan carcinoma antigen EpCAM. AB - The utility of adenoviral vectors for cancer therapy is limited due to their lack of specificity for tumor cells. In order to target adenovirus to tumor, the natural tropism of the adenovirus should be ablated and replaced by a tumor specific binding domain. To this end, a neutralizing anti-fiber antibody conjugated to an anti-EpCAM antibody was created that targets the adenovirus to the EpCAM antigen present on tumor cells. The EpCAM antigen was chosen as the target because this antigen is highly expressed on a variety of adenocarcinomas of different origin such as breast, ovary, colon and lung, whereas EpCAM expression is limited in normal tissues. In these studies, the EpCAM-targeted adenovirus was shown to infect specifically cancer cell lines of different origin expressing EpCAM such as ovary, colon and head and neck. Gene transfer was blocked by excess anti-EpCAM antibody and dramatically reduced in EpCAM negative cell lines, thus showing the specificity of the EpCAM-targeted adenovirus. Importantly, infection with targeted adenovirus was independent of CAR, which is the natural receptor for adenovirus binding, since blocking of CAR with recombinant fiber knob did not affect infection with targeted adenovirus. Apart from the cancer cell lines, the efficacy of targeted viral infection was studied in freshly isolated primary human colon cancer cells. As colon cancer predominantly metastasizes to liver, and adenovirus has a high tropism for hepatocytes, we also sought to determine if the EpCAM-targeted adenovirus showed reduced infectivity of human liver cells. The bispecific antibody could successfully mediate gene transfer to primary human colon cancer cells, whereas it almost completely abolished infection of liver cells. This work thus demonstrates that EpCAM-targeted adenoviral vectors can be specifically directed to a wide variety of adenocarcinomas. This approach may prove to be useful for selective gene therapy of cancer. PMID- 10467373 TI - pCOR: a new design of plasmid vectors for nonviral gene therapy. AB - A totally redesigned host/vector system with improved properties in terms of safety has been developed. The pCOR plasmids are narrow-host range plasmid vectors for nonviral gene therapy. These plasmids contain a conditional origin of replication and must be propagated in a specifically engineered E. coli host strain, greatly reducing the potential for propagation in the environment or in treated patients. The pCOR backbone has several features that increase safety in terms of dissemination and selection: (1) the origin of replication requires a plasmid-specific initiator protein, pi protein, encoded by the pir gene limiting its host range to bacterial strains that produce this trans-acting protein; (2) the plasmid's selectable marker is not an antibiotic resistance gene but a gene encoding a bacterial suppressor tRNA. Optimized E. coli hosts supporting pCOR replication and selection were constructed. High yields of supercoiled pCOR monomers were obtained (100 mg/l) through fed-batch fermentation. pCOR vectors carrying the luciferase reporter gene gave high levels of luciferase activity when injected into murine skeletal muscle. PMID- 10467372 TI - Anti-tumour activity against B16-F10 melanoma with a GM-CSF secreting allogeneic tumour cell vaccine. AB - Genetic modification of tumour cells with the GM-CSF encoding gene renders these cells more potent, as autologous tumour cell vaccine, than their wild-type counterparts. However, autologous vaccines are impractical for wide-scale clinical use and we have therefore investigated the efficacy of the GM-CSF genetic modification approach with an allogeneic whole cell tumour vaccine. In this report, we show that the allogeneic K1735-M2 (H-2k) melanoma cell vaccine induces a specific protective anti-tumour response against the syngeneic B16-F10 (H-2b) melanoma tumour in C57BL/6J mice. In vitro T cell work demonstrated that vaccination of animals with the allogeneic cell vaccine generated cytotoxic T cells specific for the autologous tumour. In vivo T cell subset depletion experiments also illustrated that this anti-tumour effect was mediated by both CD4+ve and CD8+ve T cells, suggesting that the allogeneic vaccine may operate through the 'cross-priming' phenomenon whereby tumour antigens are processed and presented to T cells by the host's own antigen presenting cells (APC). Thus, we transduced K1735-M2 cells with a GM-CSF expressing retroviral vector and showed anti-tumour activity of the GM-CSF secreting K1735-M2 cells as a therapeutic vaccine against the syngeneic B16-F10 tumour. Our data imply that GM-CSF genetically modified allogeneic whole cell tumour vaccines could be successful in the clinic. In addition, more potent combination gene therapy strategies could be tested using this therapeutic allogeneic vaccine model. PMID- 10467374 TI - A novel dual function retrovirus expressing multidrug resistance 1 and O6 alkylguanine-DNA-alkyltransferase for engineering resistance of haemopoietic progenitor cells to multiple chemotherapeutic agents. AB - Following transduction with a retrovirus (SF1MIH) expressing both the multidrug resistance 1 (MDR1) and O6-alkylguanine-DNA-alkyltransferase (ATase) proteins, human erythroleukaemic progenitor (K562) cells were isolated which were resistant to killing by the MDR1 substrate, colchicine. In colony-forming survival assays, K562-SF1MIH cells exhibited resistance to colchicine and doxorubicin, as well as to the O6-alkylating agents N-Methyl-N-nitrosourea (MNU) and temozolomide. Furthermore, the resistance to doxorubicin was abolished by preincubation with the MDR1 inhibitor verapamil while resistance to MNU was ablated by the specific ATase inactivator, O6-benzylguanine (O6-beG) confirming that resistance to doxorubicin and MNU was conferred by MDR1 and ATase, respectively. When K562 SF1MIH were exposed to combinations of colchicine and MNU or doxorubicin and temozolomide, simultaneous resistance to these agents was observed. Thus, transduction of K562 with SF1MIH conferred dual resistance to these cells. These data offer the prospect of designing vectors that will confer resistance to entire regimens of chemotherapy rather than just to individual components of such drug cocktails, thereby substantially increasing the efficacy of therapy. Furthermore, the use of such dual expression constructs is likely to be highly informative for the design of effective in vivo selection protocols, an issue likely to make a major impact in a clinical context in gene therapy in the near future. PMID- 10467375 TI - Scoring and databasing in multiple sclerosis. Proceedings of the European Charcot Foundation Symposium. Nice, France, 29-31 October 1998. PMID- 10467377 TI - Clinical progression and decision making process in multiple sclerosis. PMID- 10467376 TI - Databases in MS research: pitfalls and promises. AB - A database is an organized repository of data. Prospective collection of patient information in a database ('databasing') has been attempted by a few consortia of MS investigators over the past 10 years. This approach promises to facilitate epidemiologic research in MS and investigation of the natural history of the disease and how it might be altered by long-term treatments such as interferon beta. Databasing has some advantages over clinical trials in assessing new therapies, primarily because the focus is on long-term effectiveness in an entire population rather than short-term statistical significance in a highly selected population. The limitations of databasing and strategies to overcome these limitations are addressed. PMID- 10467378 TI - Quantification of impairment in MS: discussion of the scales in use. AB - Accurate, efficient measurement of disease status has become a key issue in MS clinical practice and research. Despite some perceived problems, the Expanded Disability Status Scale (EDSS) remains the most widely used comprehensive measure of impairment. There is also a place for more restricted scales of impairment which focus on some aspects of neurological function in more detail than the EDSS e.g. measures of ambulation, arm function and cognitive function. Improvements in scales of impairment in the future is likely to result from the use of more quantitative tests of selected components of the neurological examination. PMID- 10467379 TI - Use of composite scores in the quantification of deterioration in multiple sclerosis. PMID- 10467380 TI - The Guy's Neurological Disability Scale (GNDS): a new disability measure for multiple sclerosis. AB - A postal survey showed that the majority of 49 leading international neurologists involved with multiple sclerosis research felt that currently existing outcome measures for this illness were inadequate, and that there was a need for a new measure which should be patient orientated, multidimensional, and not biased towards any particular disability. The Guy's Neurological Disability Scale (GNDS) was subsequently devised as a simple and user-friendly clinical disability scale capable of embracing the whole range of disabilities which could be encountered in the course of multiple sclerosis. It has 12 separate categories which include cognition, mood, vision, speech, swallowing, upper limb function, lower limb function, bladder function, bowel function, sexual function, fatigue, and 'others'. The GNDS was found to be acceptable to neurologists and patients, reliable, responsive, and valid as a measure of disability. The scale was also found to be valid when applied by non-neurologists, over the phone, or via a postal questionnaire. PMID- 10467381 TI - Design of a European multicenter study dedicated to the evaluation of the EDMUS system: EVALUED. European Database for MUltiple Sclerosis. EVALUation of the EDMUS system. PMID- 10467382 TI - Assessment of cognitive and psychological functions in patients with multiple sclerosis: considerations for databasing. AB - Test batteries for use in studying cognition and depression in patients are briefly but critically reviewed. Suggestions for a new battery that contains tests that are related to rehabilitation and work status are proposed. PMID- 10467383 TI - The Multiple Sclerosis Functional Composite Measure (MSFC): an integrated approach to MS clinical outcome assessment. National MS Society Clinical Outcomes Assessment Task Force. AB - Clinical outcome assessment in Multiple Sclerosis (MS) is challenging due to the diversity and fluctuating nature of MS symptoms. Traditional clinical scales such as the EDSS are inadequate in their assessment of key clinical dimensions of MS (e.g. , cognitive function), and they have psychometric limitations as well. Based on analyses of pooled data from natural history studies and from placebo groups in clinical trials, the National MS Society's Clinical Outcomes Assessment Task Force recently proposed a new multidimensional clinical outcome measure, the MS Functional Composite (MSFC). The MSFC comprises quantitative functional measures of three key clinical dimensions of MS: leg function/ambulation, arm/hand function, and cognitive function. Scores on component measures are converted to standard scores (z-scores), which are averaged to form a single MSFC score. Preliminary analyses confirm that: (1) the three clinical dimensions of the MSFC are relatively independent; (2) the MSFC is sensitive to clinical changes over 1- and 2-year intervals; and (3) the MSFC has acceptable criterion validity (i.e., predicts both concurrent and subsequent EDSS change). The advantages and potential limitations of incorporating quantitative functional outcome measures such as the MSFC into collaborative databases are discussed. PMID- 10467384 TI - Recent developments in the assessment of quality of life in multiple sclerosis (MS). AB - Multiple sclerosis (MS) and its treatment have broad-ranging effects on quality of life. This article reviews recent efforts to assess the impact of MS on activities of daily living (ADLs) and health-related quality of life (HRQL), and describes the development of the Multiple Sclerosis Quality of Life Inventory (MSQLI). The MSQLI is a modular MS-specific HRQL instrument consisting of a widely-used generic measure, the Health Status Questionnaire (SF-36), supplemented by nine symptom-specific measures (covering fatigue, pain, bladder function, bowel function, emotional status, perceived cognitive function, visual function, sexual satisfaction, and social relationships). Content validation consisted of evaluating its adherence to a conceptual model of the impact of MS, and review by MS specialists (neurologists and allied health professionals), HRQL experts, patients, and caregivers. The reliability and construct validity of the MSQLI were rigorously evaluated in a field test with 300 North American patients (198 female, 102 male) with definite MS (Poser criteria) and a broad range of physical impairment (EDSS=0. 0-8.5). This article concludes by comparing the MSQLI with two other MS-specific HRQL measures (MS Quality of Life-54 (QOL-54) and Functional Assessment of Multiple Sclerosis (FAMS)) and discussing key issues to consider in selecting an HRQL instrument for a collaborative database. PMID- 10467385 TI - Measuring handicap in multiple sclerosis. AB - Handicap, shortly to be renamed participation, is a well defined though somewhat neglected entity which addresses an important aspect of the impact disease has on the individual. It is particularly relevant in Multiple Sclerosis (MS) which has such a major impact on issues such as employment, relationships, transport and social integration. Few validated measuring tools exist, and the generic London Handicap Scale is probably the best currently available but is only appropriate for group comparison. Handicap should be monitored in MS and is particularly appropriate in the evaluation of rehabilitation strategies. PMID- 10467386 TI - Measuring evoked responses in multiple sclerosis. AB - Evoked potentials (EPs) have been widely utilised in Multiple Sclerosis (MS) patients to demonstrate the involvement of sensory and motor pathways. Their diagnostic value is based on the ability to reveal clinically silent lesions and to objectivate the central nervous system damage in patients who complain frequently of vague and indefinite disturbances which frequently occurs in the early phases of the disease. The advent of magnetic resonance imaging (MRI) techniques has greatly reduced the clinical utilisation of EPs, which is not fully justifiable, as the information provided by EPs are quite different from those provided by MRI. The abnormalities of evoked responses reflect the global damage of the evoked nervous pathway and are significantly correlated with the clinical findings, while the vast majority of MRI lesions are not associated to symptoms and signs. Transversal and longitudinal studies have demonstrated that EP changes in MS are more strictly related to disability than MRI lesion burden. On the contrary, MRI is more sensitive than EPs in revealing the disease activity. Evoked responses modifications observed in MS are not disease-specific; moreover longitudinal studies showed latency and morphology changes of evoked responses not always related to clinical changes. Such a dissociation can be explained both by technical factors and by subclinical disease activity. To reduce the negative impact of technical aspects, only reproducible parameters of the evoked responses should be used to monitor disease evolution and therapeutic interventions. PMID- 10467387 TI - Magnetic resonance imaging in the monitoring of disease progression in multiple sclerosis. AB - Disease progression and irreversible disability in multiple sclerosis results from incomplete recovery from relapses, but most importantly from insidious disease progression. Although magnetic resonance imaging parameters, such as new lesion rate and gadolinium enhancement, reflect inflammation and disease activity they have no bearing on disease progression. Until now the T2 lesion load or disease burden has been relied upon for this, despite its poor relationship with disability measures. This paper looks at the mechanisms responsible for disease progression and discusses the MR techniques now available to reflect these pathological processes. PMID- 10467388 TI - Normal-appearing white matter changes in multiple sclerosis: the contribution of magnetic resonance techniques. AB - Several magnetic resonance (MR) techniques have proved to be sensitive enough to detect the subtle pathological changes that post-mortem studies showed to occur in the normal-appearing white matter (NAWM) from patients with multiple sclerosis (MS). Although these abnormalities can be detected in other neurological conditions, they seem to be more frequent and diffuse in MS. However, the contribution of NAWM changes to the diagnosis is still unclear. Their nature is also unknown and perhaps differs in different phases and clinical manifestations of the disease. Nevertheless, the extent and severity of NAWM damage seems to be relevant in causing disability and influencing the clinical evolution in MS patients. This review will summarize the present knowledge about MR-detected NAWM changes in MS and their relevance to the diagnosis and the understanding of disease evolution. PMID- 10467389 TI - MRI in multiple sclerosis: correlation with expanded disability status scale (EDSS). AB - Magnetic resonance (MR) imaging is very sensitive in showing disseminated MS lesions. Subclinical MR progression occurs frequently, explaining why MR is now used to monitor treatment, even without measurable consequences, of new MR lesions to the patient at this moment. In the light of this clinico-radiological paradox, the significance of MR in MS is discussed, particularly in relation with the expanded disability status scale (EDSS). Gadolinium-enhancing lesions correlate with the occurrence of relapses, CSF myelin breakdown products and, in patients with relapsing-remitting disease, with higher EDSS. However, the predictive value of the frequency of enhancement for changes in EDSS is only weak. For conventional T2-weighted MR imaging, the cross-sectional correlation with EDSS varies between 0.15 and 0.60, and is limited mainly by the inherent lack of tissues specificity of T2-weighted images. Both T1 black holes and magnetisation transfer (MT) parameters show a better correlation with EDSS; it should be noted that lesions in which those abnormalities are found go through an initial phase of enhancement as well. For T1 black holes, a correlation up to 0.81 has been reported for SP patients. Post-mortem studies show that black holes and low MT ratios are in vivo markers of axonal loss. Preliminary data indicate that progressive atrophy also correlates with progression on the EDSS scale. More should be learned about the fate of new MR lesion with regards to development of axonal loss, which at present is difficult to predict in the enhancing stage. The existence of escape mechanisms, including remyelination, make a simple correlation with EDSS extremely unlikely, and perhaps not even desirable. Nevertheless, while the clinical effect of a given new lesion may be difficult to ascertain, the absence of (new) MR lesions is prognostically favourable, as will be the degree to which new lesions are prevented by treatment. PMID- 10467390 TI - Biological markers in body fluids for activity and progression in multiple sclerosis. AB - Reliable biological markers in body fluids for disease activity and progression are important for our understanding of the pathophysiology and therapeutic decisions in various subtypes of multiple sclerosis. Sampling from body fluids such as cerebrospinal fluid, blood, and urine constitutes the problem that the local immuno-inflammatory process takes place in the central nervous system whereas the disease activity is only to some extent reflected in the systemic immune compartment. Promising results have been obtained in studies of adhesion molecules, pro-inflammatory cytokines, co-stimulatory molecules and neopterin as markers of disease activity in relapsing-remitting multiple sclerosis. However, these results apply to groups of patients but not necessarily to individual patients. Currently no single body fluid marker is sufficiently correlated to disease activity to be used in the individual patient in monitoring disease activity, progression, or therapeutic effects. PMID- 10467391 TI - The revision of the International Classification of Impairments, Disabilities and Handicaps and its application in MS. PMID- 10467392 TI - The Danish Multiple Sclerosis Registry: a 50-year follow-up. AB - The Danish Multiple Sclerosis Registry was established in 1948 in continuation with a nationwide survey of the prevalence of Multiple Sclerosis (MS) in Denmark. The register has since collected information on MS patients from all Danish departments of neurology, practising neurologists, MS rehabilitation centres, the National Patient Registry, the Danish MS Society, and departments of neuropathology. The registry is linked with the Danish Central Population Registry. The completeness has been estimated at more than 90%. All cases are reclassified by two neurologists as to diagnosis and year of onset. 12 070 cases with a confirmed diagnosis of MS are kept in the databases. They were prevalent in 1949 or have had onset in the period 1948 - 1993. The registry is continuously updated with new information on registered cases and new cases. The crude average annual incidence rate 1980 - 89 was 4.99/105; the prevalence rate was 112/105 by 1 January 1990. Cross-linking with other registers have enabled analytical prospective epidemiological studies, and the registry has provided population based unbiased samples of patients for a number of clinical studies. PMID- 10467393 TI - The UK Multiple Sclerosis database. PMID- 10467394 TI - MUSIS 2.0 - Multiple Sclerosis Information System: An easy-to-use-database to improve the care of patients with multiple sclerosis. PMID- 10467395 TI - The patient's role in the improvement of care. PMID- 10467396 TI - Bcl-2/E1B 19 kDa-interacting protein 3-like protein (Bnip3L) interacts with bcl 2/Bcl-xL and induces apoptosis by altering mitochondrial membrane permeability. AB - We have previously reported on cloning of the human gene encoding Bcl 2/adenovirus E1B 19 kDa-interacting protein 3-like protein (Bnip3L) and its growth inhibitory effect on cancer cells. Here we show that Bnip3L contains a motif similar to the BH3 domain which is conserved in Bcl-2 family proteins as well as containing a membrane-anchoring domain, and that Bnip3L interacts with Bcl-2 and Bcl-xL. Immunofluorescence microscopy revealed that Bnip3L was localized in the mitochondria, when in the presence of the membrane-anchoring domain. Transient expression of Bnip3L induced apoptosis of Rat-1 and HeLa cells and mutational analysis revealed that the BH3 domain and the membrane-anchoring domain were required for Bnip3L to induce cell death. Addition of recombinant Bnip3L to isolated mitochondria induced membrane potential loss and cytochrome c release both of which have been suggested to be prerequisite for apoptotic cell death. These results suggest that Bnip3L is one of the BH3-containing pro apoptotic proteins and that it targets the mitochondria when inducing apoptosis. PMID- 10467397 TI - Isolation of DICE1: a gene frequently affected by LOH and downregulated in lung carcinomas. AB - In the development and progression of sporadic tumors multiple tumor suppressor genes are inactivated that may be distinct from predisposing cancer genes. Previously, a tumor suppressor locus on human chromosome 13q14 that is distinct from the retinoblastoma predisposing gene 1 (RB1) has been identified in lung, head and neck, breast, ovarian and prostate tumors. By an approach that combines genomic difference cloning and positional cloning we isolated the cDNA of a novel gene (DICE1) located at 13q14.12-14.2. The DICE1 gene is highly conserved in evolution and its mRNA is expressed in a wide variety of fetal and adult tissues. The DICE1 cDNA encodes a predicted protein of 887 amino acids corresponding to an 100 kD protein that shows 92.9% identity to the carboxy-terminal half of the mouse EGF repeat transmembrane protein DBI-1. The DBI-1 protein interferes with the mitogenic response to insulin-like growth factor 1 (IGF-I) and is presumably involved in anchorage-dependent growth. When compared to normal lung tissue expression of the DICE1 mRNA was reduced or undetectable in the majority of non small cell lung carcinomas analysed. The location of the DICE1 gene in the region of allelic loss, its high evolutionary conservation and the downregulation of expression in carcinoma cells suggests that DICE1 is a candidate tumor suppressor gene in non-small cell lung carcinomas and possibly in other sporadic carcinomas. PMID- 10467398 TI - Papillomavirus E2 induces p53-independent apoptosis in HeLa cells. AB - We have previously shown that expression of the papillomavirus E2 protein in HeLa cells induces p53 accumulation and causes both cell cycle arrest and apoptosis. In contrast to growth arrest, onset of apoptosis was not correlated with an increase of p53 transcriptional activity. In the present study, we conducted biochemical and genetic experiments in order to determine whether E2-induced apoptosis was independent of p53 induction. We showed that E2 did not alter the transcription of Bax, a known p53-activated cell death inducer. The time course of apoptotic cell death preceded p53 induction by several hours. Overexpression of the HPV18 E6 oncogene prevented E2-mediated p53 accumulation, but did not alter the rate of cell death. Finally, point mutants of the HPV18 E2 transactivation domain induced apoptosis, although they were unable to induce high p53 accumulation or cell cycle arrest. In addition, the results obtained with these mutants indicated that both transcriptional activation and replication functions of E2 were dispensable for the induction of cell death. These observations show that E2-induced apoptosis is an early event, independent of p53 accumulation and unrelated to downstream p53-dependent transcriptional events. PMID- 10467399 TI - Cloning and characterization of the human BAG-1 gene promoter: upregulation by tumor-derived p53 mutants. AB - BAG-1 is an anti-apoptotic protein that interacts with Bcl-2, Bcl-XL, Hsp70/Hsc70, Raf-1 and numerous hormone or growth factor receptors. Recently, BAG 1 has been found to be overexpressed in a variety of human cancer cell lines and some tumors. However, the molecular mechanism of BAG-1 upregulation is still unclear. In this study, we cloned 0.9 kb of human genomic DNA, BGEV, 5' flanking the BAG-1 open reading frame. BGEV subcloned into a promoterless luciferase reporter vector conferred high promoter activity in various human cancer cell lines. Deletion analysis of this sequence localized the region of maximal BAG-1 promoter activity from nucleotide positions -353 to -54, upstream of the first start codon CTG. Sequence analysis of the BAG-1 promoter region showed the absence of a TATA box but identified a CCAAT box, several GC boxes, a CpG island and several transcriptional factor binding sites, which may be important in the regulation of BAG-1 transcription. Most importantly, functional characterization of the BAG-1 promoter in vivo demonstrated that gain-of-function p53 mutants derived from human tumors upregulated the transcription of BAG-1 RNA and the expression of a reporter gene from the BAG-1 promoter. These results indicated that we have isolated the functional constitutive BAG-1 promoter. Furthermore, the data suggested that overexpression of BAG-1 in some tumors may be due to upregulation of the human BAG-1 promoter by mutant p53. PMID- 10467400 TI - Overexpression of urokinase-type plasminogen activator in pancreatic adenocarcinoma is regulated by constitutively activated RelA. AB - The Rel/NF-kappaB transcription factors regulate the expression of many genes. The activity of RelA, a member of the Rel/NF-kappaB transcription factor family, is constitutively activated in the majority of pancreatic adenocarcinomas and cell lines. We report that the urokinase-type plasminogen activator (uPA), one of the critical proteases involved in tumor invasion and metastasis, is overexpressed in pancreatic tumor cells and its overexpression is induced by constitutive RelA activity. The uPA promoter contains an NF-kappaB binding site that directly mediates the induction of uPA expression by RelA. Expression of a dominant-negative IkappaBalpha mutant inhibits kappaB site-dependent transcriptional activation of a uPA promoter-CAT reporter gene. Treating the pancreatic tumor cell lines with the known NF-kappaB inhibitors, dexamethasone and n-tosylphenyalanine chloromethyl ketone (TPCK), abolishes constitutive RelA activity and uPA overexpression. These results show that uPA is one of the downstream target genes induced by constitutively activated RelA in human pancreatic tumor cells, and suggests that constitutive RelA activity may play a critical role in tumor invasion and metastasis. Inhibition of constitutive RelA in pancreatic tumor cells may reduce their invasive and metastatic potential. PMID- 10467401 TI - Induction of mammary gland hyperplasia in transgenic mice over-expressing human Cdc25B. AB - Cdc25 A and B are dual-specificity phosphatases which have been implicated in neoplastic transformation. Although Cdc25A and Cdc25B have been found to be over expressed in many cancer cell lines and primary tumors, the physiological roles of Cdc25A and B in vivo are largely undefined. To investigate the roles of these proteins in the oncogenic transformation of the mammary gland we used the mouse mammary tumor virus (MMTV) promoter to target over-expression of the Cdc25B transgene in the mammary glands of transgenic mouse lines. Here we report that the over-expression of Cdc25B enhances the proliferation of mammary epithelial cells resulting in the formation of precocious alveolar hyperplasia. At the molecular level, marked increases in cyclin D1 protein have been found in transgenic mammary epithelial cells. The accelerated growth rate of the mammary epithelial cells could also be attributed to the increased levels of cyclin E/cdk2 activity. In addition, a pronounced decrease in apoptosis was also observed during the involution of mammary gland. The reduction of apoptosis during involution correlated well with the reduced expression of c-myc and p53, both of which have been implicated in apoptosis. Taken together, our results clearly indicate that the deregulated expression of Cdc25B generates mammary gland hyperplasia. PMID- 10467402 TI - Differential expression of the cyclin-dependent kinase inhibitor P27 in primary hepatocytes in early-mid G1 and G1/S transitions. AB - P27, an inhibitor of cyclin-dependent kinases, plays an important role in the control of cell adhesion and contact inhibition-dependent cell cycle regulation. Hepatocytes, maintained in primary culture, offer a model of synchronized primary epithelial cells which retain a differentiated profile while stimulated to proliferate. We therefore investigated the pattern of endogenous p27 expression in cyclin rat hepatocytes isolated by collagenase perfusion followed by mitogenic stimulation. P27 was expressed in whole normal liver and freshly isolated hepatocytes. We then observed a sharp decrease in p27 levels, concomitant with the progression in early-mid G1, followed by reaccumulation in late G1 and the G1/S transition. Immunochemistry and BrdU labelling demonstrated nuclear localization of p27 and its expression in cells engaged in both G1 and S phase. P27 was detected in late G1 in complexes containing cyclins D1, E and A. Cyclin E and A-associated kinase activities, however, were detected at the G1/S transition and depletion experiments confirmed that most active complexes were free of p27. Phosphorylated forms of p27 were detected in unstimulated and stimulated hepatocytes in both early-mid G1 and G1/S. Finally, two-dimensional gel electrophoresis showed evidence for several forms of p27 with a distinct profile of distribution in quiescent and stimulated hepatocytes. Collectively, our data offer a model in which p27 shows a biphasic profile of accumulation, with the early decrease possibly involved in the progression through early and mid G1. In contrast with most cell types tested so far, the late G1 accumulation did not impair formation of active cyclin E- and A associated kinases, and thus G1/S transition. PMID- 10467403 TI - The selective and inducible activation of endogenous PI 3-kinase in PC12 cells results in efficient NGF-mediated survival but defective neurite outgrowth. AB - The Trk/Nerve Growth Factor receptor mediates the rapid activation of a number of intracellular signaling proteins, including phosphatidylinositol 3-kinase (PI 3 kinase). Here, we describe a novel, NGF-inducible system that we used to specifically address the signaling potential of endogenous PI 3-kinase in NGF mediated neuronal survival and differentiation processes. This system utilizes a Trk receptor mutant (Trk(def)) lacking sequences Y490, Y785 and KFG important for the activation of the major Trk targets; SHC, PLC-gammal, Ras, PI 3-kinase and SNT. Trk(def) was kinase active but defective for NGF-induced responses when stably expressed in PC12nnr5 cells (which lack detectable levels of TrkA and are non-responsive to NGF). The PI 3-kinase consensus binding site, YxxM (YVPM), was introduced into the insert region within the kinase domain of Trk(def). NGF stimulated tyrosine phosphorylation of the Trk(def)+PI 3-kinase addback receptor, resulted in the direct association and selective activation of PI 3-kinase in vitro and the production of PI(3,4)P2 and PI(3,4,5)P3 in vivo (comparable to wild type). PC12nnr5 cells stably expressing Trk(def) + PI 3-kinase, initiated neurite outgrowth but failed to stably extend and maintain these neurites in response to NGF as compared to PC12 parental cells, or PC12nnr5 cells overexpressing wild type Trk. However, Trk(def) + PI 3-kinase was fully competent in mediating NGF induced survival processes. We propose that while endogenous PI 3-kinase can contribute in part to neurite initiation processes, its selective activation and subsequent signaling to downstream effectors such as Akt, functions mainly to promote cell survival in the PC12 system. PMID- 10467404 TI - Transcriptional regulation by targeted recruitment of cyclin-dependent CDK9 kinase in vivo. AB - The CDK9 kinase in association with Cyclin T is a component of the transcription positive-acting complex pTEFb which facilitates the transition from abortive to productive transcription elongation by phosphorylating the carboxyl-terminal domain of RNA polymerase II. The Cyclin T1/CDK9 complex is implicated in Tat transactivation, and it has been suggested that Tat functions by recruiting this complex to RNAPII through cooperative binding to RNA. Here, we demonstrate that targeted recruitment of Cyclin T1/CDK9 kinase complex to specific promoters, through fusion to a DNA-binding domain of either Cyclin T1 or CDK9 kinase, stimulates transcription in vivo. Transcriptional enhancement was dependent on active CDK9, as a catalytically inactive form had no transcriptional effect. We determined that, unlike conventional activators, DNA-bound CDK9 does not activate enhancerless TATA-promoters unless TBP is overexpressed, suggesting that CDK9 acts in vivo at a step subsequent to TFIID recruitment DNA-bound. Finally, we determined that CDK9-mediated transcriptional activation is mediated by preferentially stimulating productive transcription elongation. PMID- 10467405 TI - A role of cyclin G in the process of apoptosis. AB - Cyclin G was previously identified as a target gene of the p53 tumor suppresser protein, and levels of cyclin G are increased after induction of p53 by DNA damage. However, the function of cyclin G has not been established. To determine the effect of increased expression of cyclin G, retroviruses encoding cyclin G were constructed and used to infect three different murine cell lines. Cyclin G protein levels induced by the retroviruses were within the range seen after DNA damage induction of p53. In each case we observed that such over-expression of cyclin G augments the apoptotic process. TNF-alpha induction of apoptosis is increased by expression of cyclin G in NIH3T3 fibroblasts which express p53, as well as in 10.1 fibroblasts which contain no p53 allele. Additionally, we observed that while cyclin G expression is markedly reduced upon aggregate formation in embryonic carcinoma P19 cells, retrovirus-mediated over-expression of cyclin G enhances apoptotic cell death in aggregated P19 cells, and increases the extent of apoptosis caused by retinoic acid or serum starvation of these cells. These data demonstrate that cyclin G plays a facilitating role in modulating apoptosis induced by different stimuli. Moreover, we have discovered that cyclin G expression is rapidly induced in P19 cells after exposure to Bone Morphogenic Protein-4 (BMP-4), suggesting that cyclin G may mediate apoptotic signals generated by BMP-4. PMID- 10467406 TI - Suppression of the poly(ADP-ribose) polymerase activity by DNA-dependent protein kinase in vitro. AB - It has been suggested that DNA-dependent protein kinase (DNA-PK) is a central component of DNA double-strand-break repair. The mechanism of DNA-PK action, however, has not been fully understood. Poly(ADP-ribose) polymerase (PARP) is another nuclear enzyme which has high affinity to DNA ends. In this study, we analysed the interaction between these two enzymes. First, DNA-PK was found to suppress the PARP activity and alters the pattern of poly(ADP-ribosyl)ation. Although DNA-PK phosphorylates PARP in a DNA-dependent manner, this modification is unlikely to be responsible for the suppression of PARP activity, since this suppression occurs even in the absence of ATP. Conversely, PARP was found to ADP ribosylate DNA-PK in vitro. However, the auto-phosphorylation activity of DNA-PK was not influenced by this modification. In a competitive electrophoretic mobility shift assay, Ku 70/80 complex, the DNA binding component of DNA-PK, was found to have higher affinity to a short fragment of DNA than does PARP. Furthermore, co-immunoprecipitation analysis suggested direct or close association between Ku and PARP. Thus, DNA-PK suppresses PARP activity, probably through direct binding and/or sequestration of DNA-ends which serve as an important stimulator for both enzymes. PMID- 10467407 TI - c-Myc does not prevent glucocorticoid-induced apoptosis of human leukemic lymphoblasts. AB - Due to their growth arrest- and apoptosis-inducing ability, glucocorticoids (GC) are widely used in the therapy of various lymphoid malignancies. The signal transduction pathways leading to this clinically-relevant form of apoptosis have, however, not been sufficiently elucidated. GC bind to their specific receptor, a ligand-activated transcription factor of the Zn-finger type, that activates or represses transcription of GC-responsive genes. Previous studies in leukemia cells suggested that transcriptional repression of c-myc expression might be the crucial event in GC-induced apoptosis, although in other systems, c-Myc apparently increased the sensitivity to cell-death inducers. To address this controversy, we stably transfected the GC-sensitive human T-ALL cell line CEM C7H2 with constructs allowing tetracycline-regulated expression of c-Myc. Subsequent analyses of these cell lines showed that overexpression of c-Myc per se had little, if any, effect on cell viability, although it rendered the cells more sensitive to apoptosis induced by low serum, confirming the functionality of the expressed transgene. More importantly, however, when the cells were treated with GC in the presence of exogenous c-Myc, they underwent apoptosis exceeding that in cells treated in the absence of transgenic c-Myc. The data indicate that c-myc downregulation is not critical for induction of cell-death by GC in this system, and support the notion that c-Myc sensitizes cells to apoptosis-inducing agents. PMID- 10467408 TI - Synergy of PEBP2/CBF with mi transcription factor (MITF) for transactivation of mouse mast cell protease 6 gene. AB - The mi locus encodes a member of the basic - helix - loop - helix - leucine zipper (bHLH-Zip) protein family of transcription factors (hereafter called MITF). Although the bHLH-Zip family transcription factors generally recognize and bind CANNTG motifs, the expression of mouse mast cell protease 6 (MMCP-6) gene is regulated by MITF through the GACCTG motif in the promoter region. The GACCTG motif was partly overlapped the TGTGGTC sequence, which was bound by polyomavirus enhancer binding protein 2 (PEBP2). In the present study, the effect of PEBP2 on the expression of MMCP-6 gene was examined. PEBP2 that is composed of alpha and beta subunits was expressed by mast cell lines and cultured mast cells derived from spleen. The overexpression of dominant negative PEBP2 cDNA reduced the expression of MMCP-6. Moreover, the simultaneous transfection of the plasmid containing MITF cDNA and the plasmid containing PEBP2 cDNA increased the MMCP-6 promoter activity. For the synergistic action of PEBP2 and MITF, the intact GACCTG and TGTGGTC motifs were prerequisite. The PEBP2alphaB1 mutant which lacked the region downstream from the Runt domain did not bind MITF and lost the synergistic function. These results indicated that PEBP2 and MITF synergistically transactivated the MMCP-6 gene and that the region downstream from the Runt domain of PEBP2alphaB1 was essential for the physical and functional interactions with MITF. PMID- 10467409 TI - Frequent loss of heterozygosity at 1p36 in ovarian adenocarcinomas but the gene encoding p73 is unlikely to be the target. AB - Loss of heterozygosity (LOH) involving the distal part of the short arm of chromosome 1 occurs frequently in ovarian adenocarcinomas but the tumour suppressor gene(s) targeted by this event is unknown. We have used five microsatellite markers in a panel of 56 ovarian adenocarcinomas to determine which part of 1p34 - 36 is the focus of this LOH. LOH was considerably more common at 1p36 (43%) than at 1p34 - 35 (18%), and 11 tumours showed LOH at 1p36 but not at 1p34 - 35. These data strongly suggest the presence of a tumour suppressor gene inactivated in ovarian adenocarcinoma at 1p36. The p53 homologue, p73, has recently been isolated and mapped to 1p36 and therefore is a candidate for this tumour suppressor gene. However, RT - PCR and Western analyses revealed strong expression of p73 in ovarian adenocarcinoma cell lines but very low or undetectable levels in normal ovarian surface epithelial cells. Immunohistochemical analysis of primary ovarian tumours showed that only 3/22 (14%) contained p73 expressing cells. There was no association between 1p36 LOH and p73 expression in ovarian tumours, nor between p73 and p53 expression. These findings strongly suggest that p73 is not the target of 1p36 LOH in ovarian adenocarcinomas but indicate the presence of an, as yet unidentified, tumour suppressor gene in this region that plays an important role in ovarian tumorigenesis. PMID- 10467410 TI - Somatic mutations in the BRCA1 gene in Chinese sporadic breast and ovarian cancer. AB - Inherited mutations in the BRCA1 gene confer increased susceptibility to breast and ovarian cancer. Its role in sporadic carcinogenesis is not well defined. Somatic mutations in breast cancers have not been reported and to date there are only three reports of somatic mutations in sporadic ovarian cancers. To investigate the contribution of BRCA1 mutations to sporadic breast and ovarian cancer in the Chinese population, we analysed 62 samples from Chinese women using the protein truncation test. There were 40 cases of breast cancer under age 50 and 22 cases of ovarian cancer, all unselected for family history. There was no age selection for the ovarian cancers. We found two somatic BRCA1 mutations in exon 11, one in a breast cancer and the other in an ovarian cancer, both of which result in truncated proteins. Our results indicate that somatic BRCA1 mutations, like somatic mutations in the BRCA2 gene, though very rare, can be found in both breast and ovarian cancers and support a tumor suppressor function for BRCA1 in sporadic tumors. PMID- 10467411 TI - Evidence for SH3 domain directed binding and phosphorylation of Sam68 by Src. AB - Sam68 is a 68 kDa protein that associates with and is phosphorylated by the c-Src kinase at mitosis. It contains a KH domain implicated in RNA binding and several proline-rich motifs that resemble known SH3 binding sites. The SH3 domains of c Src, phosphatidylinositol 3-OH kinase, phospholipase C-gamma and Grb2 protein (containing two SH3 domains), but not other SH3 domains tested, were capable of binding Sam68 in vitro. Synthetic peptides corresponding to the proline motifs of Sam68 inhibited with different efficiencies the binding of SH3 domains to Sam68 suggesting that the proline motifs of Sam68 function as specific SH3 domain binding sites. Mutation of Sam68 SH3 binding sites further indicated that the SRC SH3 domain mediates binding of Src to unphosphorylated Sam68. Phosphorylation of Sam68 by Src kinase was inhibited when the Src SH3 binding site of Sam68 was mutated or when corresponding peptides were added to in vitro kinase reactions indicating that binding of the Src SH3 domain to a specific site near the amino terminus of Sam68 (including residues 38 - 45: PPLPHRSR) facilitates phosphorylation of Sam68 by the Src kinase domain. Sam68-based proline peptides had no effect on the phosphorylation of another in vitro substrate of Src, enolase. These results suggest that Src effectively mounts Sam68 through its SH3 domain, possibly as a mechanism to position the kinase domain close to substrate tyrosine residues in the carboxyl-half of the protein. PMID- 10467412 TI - Inhibition of pp60c-Src reduces Bcl-XL expression and reverses the transformed phenotype of cells overexpressing EGF and HER-2 receptors. AB - Tumors that overexpress HER-2/neu receptor or exhibit enhanced EGFR signaling have been reported to possess constitutively activated Src family kinases, especially pp60c-Src. High levels of pp60c-Src activity have also been reported for cell lines that overexpress the EGFR or the chimeric EGFR-HER-2 receptor. It has therefore been suggested that Src kinases may contribute significantly to the oncogenic phenotype of these cells and to the degree of malignancy of tumors that overexpress EGFR family receptors. In this study we show that the induced expression of c-SRC antisense RNA or the application of a selective Src kinase inhibitor induces growth arrest, programmed cell death and reverses the transformed properties of cells that overexpress EGFR or HER-2 receptors. We show that inhibition of Src kinase expression or activity results in the reduction of Stat3 tyrosine phosphorylation, decline of Bcl-XL expression, and induction of cell death. Using a construct in which the promoter of Bcl-X, which possesses putative Stat3 sites, is tethered to the luciferase reporter gene, we show that inhibition of Src activity or expression induces a decline in Bcl-X expression. We also show that the expression of activated Src induces activation of the Bcl-X promoter. This activation is inhibited by the expression of kinase dead Src or of Stat3beta, the dominant-negative form of Stat3. Taken together, these results support the hypothesis that Src positively regulates the transformed phenotype of cells overexpressing EGFR family kinases. Furthermore, these results also suggest that Src positively regulates Bcl-XL expression via Stat3 activation and thus acts not only as a potent mitogenic signaling element, but also as an anti apoptotic signaling protein. The combination of both activities probably confers upon activated Src its oncogenic activity. Since Src kinase is activated in many tumors, pp60c-Src kinase inhibitors may prove useful as anti-cancer agents for many types of cancer. PMID- 10467413 TI - A DNA damage repair mechanism is involved in the origin of chromosomal translocations t(4;11) in primary leukemic cells. AB - Some chromosomal translocations involved in the origin of leukemias and lymphomas are due to malfunctions of the recombinatorial machinery of immunoglobulin and T cell receptor-genes. This mechanism has also been proposed for translocations t(4;11)(q21;q23), which are regularly associated with acute pro-B cell leukemias in early childhood. Here, reciprocal chromosomal breakpoints in primary biopsy material of fourteen t(4;11)-leukemia patients were analysed. In all cases, duplications, deletions and inversions of less than a few hundred nucleotides indicative of malfunctioning DNA repair mechanisms were observed. We concluded that these translocation events were initiated by several DNA strand breaks on both participating chromosomes and subsequent DNA repair by 'error-prone-repair' mechanisms, but not by the action of recombinases of the immune system. PMID- 10467414 TI - The p53 tumor suppressor protein reduces point mutation frequency of a shuttle vector modified by the chemical mutagens (+/-)7, 8-dihydroxy-9,10-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene, aflatoxin B1 and meta-chloroperoxybenzoic acid. AB - p53 has been postulated to be the guardian of the genome. However, results supporting the prediction that point mutation frequencies are elevated in p53 deficient cells either have not been forthcoming or have been equivocal. To analyse the effect of p53 on point mutation frequency, we used the supF gene of the pYZ289 shuttle vector as a mutagenic target. pYZ289 was treated in vitro by ultraviolet irradiation, aflatoxin B1, (+/-)7,8-dihydroxy-9, 10-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene and meta-chloroperoxybenzoic acid and then transfected into p53-deficient cells with or without a p53 expression vector. p53 reduced the mutant frequency up to fivefold when pYZ289 was treated with aflatoxin B1, (+/ )7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene or meta chloroperoxybenzoic acid but not when it was ultraviolet-irradiated. The p53 dependent mutation frequency reduction was higher at a higher level of premutational lesions for aflatoxin B1 and (+/-)7,8-dihydroxy-9,10-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene and at a lower level of lesions for meta chloroperoxybenzoic acid. This suggests that the chemical mutagens produce, in a dose-dependent fashion, two kinds of DNA damage, one subject to p53-dependent mutation frequency reduction and the other not. These results indicate that p53 can reduce the point mutation frequency in a shuttle vector treated by chemical mutagens and suggest that p53 can act as guardian of the genome for at least some kinds of point mutations. PMID- 10467415 TI - Drug-induced apoptosis is delayed and reduced in XPD lymphoblastoid cell lines: possible role of TFIIH in p53-mediated apoptotic cell death. AB - The tumor suppressor gene product p53 can bind to and inhibit the helicase activity of the multisubunit transcription-repair factor TFIIH. We previously reported that p53-mediated apoptosis is attenuated in primary human fibroblasts from individuals with Xeroderma Pigmentosum (XP) that harbor mutations in the TFIIH DNA helicases XPD or XPB. In this study we show that apoptosis is reduced and delayed in three XPD lymphoblastoid cell lines (LCLs), but not in an XPD heterozygote LCL, after exposure to doxorubicin, a DNA-damaging agent and topoisomerase II inhibitor frequently used in cancer therapy. Apoptosis was assessed by quantitation of Annexin V binding to exposed phosphatidylserine residues and by caspase-mediated cleavage of Poly(ADP)Ribose Polymerase (PARP). Apoptosis induced by doxorubicin was suppressed in LCLs retrovirally transduced with the Human Papillomavirus 16 E6 oncoprotein, consistent with the hypothesis that this is a p53-dependent process. PARP cleavage was not delayed in XPD LCLs in response to anti-Fas (CD95) antibody-mediated apoptosis, thus, the defect in the apoptotic pathway in these cells lies upstream of caspase activation. Similar changes in the expression of apoptosis-effector genes, p53, and p53-responsive genes p21Cip1/WAF-1/Sid1 (p21), gadd45, bcl-2 and bax were observed in normal and XPD LCLs after treatment with doxorubicin, indicating that delayed apoptosis was not a consequence of defective transcription of these genes. Thus, our studies provide further support to the hypothesis that XPD and p53 can functionally interact in a p53-mediated apoptotic pathway. PMID- 10467416 TI - Tumor suppression by p27Kip1 and p21Cip1 during chemically induced skin carcinogenesis. AB - p27Kip1 and p21Cip1 are cyclin dependent kinase inhibitors which can arrest cell proliferation and p27 is a tumor suppressor gene. To address the mechanism of tumor suppression by p27 and to determine if p21 has a tumor suppressor phenotype, we utilized the two stage skin carcinogenesis model on p27 and p21 knockout mice. In this model, initiation, which involves mutation of H-ras induced by DMBA, can be distinguished from promotion induced by TPA, and progression to carcinoma. The mean number of papillomas did not differ between p27-/- and control littermates, but papilloma growth rate was increased and carcinomas developed earlier. Thus, p27 deficiency did not enhance initiation, but resulted in more rapid clonal expansion of initiated cells during promotion. TPA treatment reduced p27 expression in keratinocytes also supporting a role for p27 during promotion. Tumors from p27-/- mice contained mutant H-ras indicating that p27 deficiency did not substitute for mutant ras and further, that during ras driven tumor growth, p27 is partially antagonistic since its removal led to faster growth. The treated p27-/- mice also developed intestinal adenomas. p21-/- mice did not display a significant increase in tumor numbers, growth rate or progression to carcinomas and these tumors also had mutated H-ras. Carcinomas from p21-/- mice were more poorly differentiated with a high frequency of anaplastic spindle cell carcinomas. Thus p21 deficiency mainly resulted in higher grade undifferentiated tumors. PMID- 10467417 TI - Anti-apoptotic activity of p53 maps to the COOH-terminal domain and is retained in a highly oncogenic natural mutant. AB - The tumour suppressor p53 plays a complex role in the regulation of apoptosis. High levels of wild type p53 potentiate the apoptotic response, while physiological range, low levels of the protein have an anti-apoptotic activity in serum starved immortalized fibroblasts. Here we report that primary fibroblast like cells that show normal growth control are also efficiently protected from apoptosis by the endogenous p53 activity. The capacity to inhibit apoptosis is not restricted to the wild type protein: the R-->H175 p53 mutant fully retains the anti-apoptotic activity of the wild type p53, providing a possible explanation for its high oncogenicity. Using a series of point and deletion mutants of p53 under the control of tetracycline-regulated promoter we show that certain mutants, like the wild type, protect cells at low levels but lead to apoptosis when overexpressed. This latter effect is lost upon deletion of a proline-rich domain in the NH2 part of the protein. The anti-apoptotic activity can be mapped to the extreme carboxy-terminal part of the protein and is therefore independent of other well characterized p53 activities. Our results add a new level of complexity to the network of interactions mediated by p53 in normal physiology and pathology. PMID- 10467418 TI - Cloning of breakpoints in and downstream the IGF2 gene that are associated with overexpression of IGF2 transcripts in colorectal tumours. AB - The human IGF2 gene belongs to a group of imprinted genes clustered on the short arm of chromosome 11, band p15.5. It contains 9 exons and spans over 30 kb. IGF2 mRNA overexpression has been reported in human tumours and in some inherited growth disorders. It was recently demonstrated that IGF2 mRNA overexpression contributes to tumour progression and that loss of parental imprinting as well as altered transcription factors are contributing to this overexpression. We have reported structural alterations in the 3' region of the IGF2 gene in two colorectal tumours that overexpressed the IGF2 transcript by 200- and 800-fold. We cloned by the vectorette-PCR strategy, genomic DNA fragments containing the breakpoints from these tumours. The sequencing of these fragments positioned the breakpoint 2 kb downstream the IGF2 gene in one tumour, and in exon 9 in the second. Both breakpoints occurred in regions containing repetitive elements: a TGGA repeat we have identified downstream the gene, and the (CA)n repetition in exon 9. We hypothesize that a negative regulatory element, located downstream the IGF2 gene, has been deleted following these structural alterations and leads to IGF2 gene overexpression. PMID- 10467419 TI - RalA requirement for v-Src- and v-Ras-induced tumorigenicity and overproduction of urokinase-type plasminogen activator: involvement of metalloproteases. AB - Overproduction of urokinase-type plasminogen activator (uPA) and metalloproteases (MMPs) is strongly correlated with tumorigenicity and with invasive and metastatic phenotypes of human and experimental tumors. We demonstrated previously that overproduction of uPA in tumor cells is mediated by a phospholipase D (PLD)- and protein kinase C-dependent mechanism. The oncogenic stimulus of v-Src and v-Ras results in the activation of PLD, which is dependent upon the monomeric GTPase RalA. We have therefore investigated whether RalA plays a role in uPA and MMP overproduction that is observed in response to oncogenic signals. We report here that NIH3T3 cells transformed by both v-Src and v-Ras, constitutively overproduce uPA and that expression of a dominant negative RalA mutant (S28N) blocks overproduction of uPA in both the v-Src-and v-Ras transformed cells. v-Src and v-Ras also induced an upregulation of the activity of MMP-2 and MMP-9 as detected by zymograms, however only the v-Src induction correlated with MMP protein levels detected by Western blot analysis. The dominant negative RalA mutant blocked increased MMP-2 and 9 overproduction induced by v-Src, but not the increased activity of MMP-2 and 9 induced by v-Ras. And, consistent with a role for the RalA/PLD pathway in mitogenesis and tumor development, the dominant negative RalA mutant completely blocked tumor formation by v-Src- and v-Ras-transformed NIH3T3 cells injected subcutaneously in syngeneic mice. The data presented here implicate RalA and PLD as signaling mediators for tumor formation and protease production by transformed cells. PMID- 10467420 TI - Mutation of beta-catenin is an early event in chemically induced mouse hepatocellular carcinogenesis. AB - beta-catenin activation, and subsequent upregulation of Wnt-signaling, is an important event in the development of certain human and rodent cancers. Recently, mutations in the beta-catenin gene in the region of the serine-threonine glycogen kinase (GSK)-3beta phosphorylation target sites have been identified in hepatocellular neoplasms from humans and transgenic mice. In this study we examined 152 hepatocellular neoplasms from B6C3F1 mice included in five chemical treatment groups and controls for mutations in the beta-catenin gene. Twenty of 29 hepatocellular neoplasms from mice treated with methyleugenol had point mutations at codons 32, 33, 34 or 41, sites which are mutated in colon and other cancers. Likewise, nine of 24 methylene chloride-induced hepatocellular neoplasms and 18 of 42 oxazepam-induced neoplasms exhibited similar mutations. In contrast, only three of 18 vinyl carbamate-induced liver tumors, one of 18 TCDD-induced liver tumors, and two of 22 spontaneous liver neoplasms had mutations in beta catenin. Thus, there appears to be a chemical specific involvement of beta catenin activation in mouse hepatocellular carcinogenesis. Expression analyses using Western blot and immunohistochemistry indicate that beta-catenin protein accumulates along cell membranes following mutation. The finding of mutations in both adenomas and carcinomas from diverse chemical treatment groups and the immunostaining of beta-catenin protein in an altered hepatocellular focus suggest that these alterations are early events in mouse hepatocellular carcinogenesis. PMID- 10467421 TI - Transforming growth factor-beta regulates Kit ligand expression in rat ovarian surface epithelial cells. AB - In preparation for ovulation, paracrine communication between the preovulatory follicle and overlying theca/stromal cells and ovarian surface epithelium (OSE) must take place to facilitate the degradative and apoptotic events associated with ovulation. Kit tyrosine kinase receptors and their ligand, kit ligand (KL) are expressed within ovarian follicles, and ligand-induced receptor activation appears to account for some of the cell - cell interactions important for oocyte development. We investigated the expression of Kit receptors and KL in OSE cells and the possibility that modulation of their expression could affect OSE cell activity. KL mRNA and protein were detected in the OSE cell layer of rat ovaries, and primary cultures of rat OSE as well as the immortalized rat OSE cell line, ROSE 199, expressed KL, but not Kit receptors. Both primary and immortalized OSE cells preferentially expressed KL-1, rather than KL-2, transcripts, suggesting that these cells produce predominantly the soluble form of KL. Activation of the cAMP signalling pathway using dibutyryl cAMP decreased proliferation of ROSE 199 cells and elicited a threefold increase in KL expression. TGF-beta similarly inhibited ROSE 199 cell proliferation, but strongly inhibited dibutyryl cAMP induced KL expression, indicating that changes in KL expression were not directly associated with OSE cell proliferation. The expression of mostly soluble KL in the surface epithelium suggests that this cytokine may be acting in a paracrine fashion, perhaps interacting with nearby Kit receptor-bearing theca cells. PMID- 10467422 TI - Distinct expression patterns and transforming properties of multiple isoforms of Ost, an exchange factor for RhoA and Cdc42. AB - A search for transforming genes expressed in brain led to the identification of a novel isoform of Ost, an exchange factor for RhoA and Cdc42. In addition to the Dbl-homology (DH) and pleckstrin-homology (PH) domains identified in the original Ost, this isoform contained a SH3 domain and a novel HIV-Tat related (TR) domain. The presence or absence of these domains in Ost defined multiple isoforms of the protein. RT - PCR and in situ hybridization analysis revealed that these isoforms were generated by tissue-specific and developmentally restricted alternative splicing events. Whereas deletion of the N-terminus activated the transforming properties of Ost, the presence of the SH3 domain reduced the transforming activity of the protein. This inhibition was relieved by the presence of a TR domain, which contained a potential SH3 ligand sequence. The transforming activity of all Ost isoforms was inhibited by dominant negative forms of the Rho family proteins. Expression of Ost isoforms potently induced the formation of actin stress fibers and filopodia as well as JNK activity and AP1- and SRF regulated transcriptional pathways. Ost transfectants also displayed elevated levels of cyclins A and D1, suggesting that the de-regulation of these cyclins is linked to Ost-mediated transformation. PMID- 10467423 TI - Dominant negative EGFR-CD533 and inhibition of MAPK modify JNK1 activation and enhance radiation toxicity of human mammary carcinoma cells. AB - Exposure of MDA-MB-231 human mammary carcinoma cells to an ionizing radiation dose of 2 Gy results in immediate activation and Tyr phosphorylation of the epidermal growth factor receptor (EGFR). Doxycycline induced expression of a dominant negative EGFR-CD533 mutant, lacking the COOH-terminal 533 amino acids, in MDA-TR15-EGFR-CD533 cells was used to characterize intracellular signaling responses following irradiation. Within 10 min, radiation exposure caused an immediate, transient activation of mitogen activated protein kinase (MAPK) which was completely blocked by expression of EGFR-CD533. The same radiation treatment also induced an immediate activation of the c-Jun-NH2-terminal kinase 1 (JNK1) pathway that was followed by an extended rise in kinase activity after 30 min. Expression of EGFR-CD533 did not block the immediate JNK1 response but completely inhibited the later activation. Treatment of MDA-TR15-EGFR-CD533 cells with the MEK1/2 inhibitor, PD98059, resulted in approximately 70% inhibition of radiation induced MAPK activity, and potentiated the radiation-induced increase of immediate JNK1 activation twofold. Inhibition of Ras farnesylation with a concomitant inhibition of Ras function completely blocked radiation-induced MAPK and JNK1 activation. Modulation of EGFR and MAPK functions also altered overall cellular responses of growth and apoptosis. Induction of EGFR-CD533 or treatment with PD98059 caused a 3-5-fold increase in radiation toxicity in a novel repeated radiation exposure growth assay by interfering with cell proliferation and potentiating apoptosis. In summary, this data demonstrates that both MAPK and JNK1 activation in response to radiation occur through EGFR-dependent and independent mechanisms, and are mediated by signaling through Ras. Furthermore, we have demonstrated that radiation-induced activation of EGFR results in downstream activation of MAPK which may affect the radiosensitivity of carcinoma cells. PMID- 10467424 TI - Defect in long-term activation of phosphatidylinositol 3-kinase by insulin in vivo: studies in insulin-resistant hHTg rats. AB - OBJECTIVE: To study the regulation of phosphatidylinositol (PI) 3-kinase by insulin in vivo in hereditary hypertriglyceridemic and insulin resistant rat (hHTg). METHODS: Total and insulin receptor substrate-1 (IRS-1) associated PI 3 kinase activities were measured in skeletal muscles and adipose tissue after an intense insulin induced glucose utilization as accomplished by 90 min euglycemic hyperinsulinemic clamp. RESULTS: In quadriceps femoris muscle, no stimulation of total or IRS-1 associated PI 3-kinase activities was found after hyperinsulinemia in both hHTg and control rats. In contrast, in soleus muscle of control rats total PI 3-kinase activity was stimulated by insulin (P<0.001), while any such effect was not found in hHTg rats. IRS-1 associated PI 3-kinase activity in soleus muscle was significantly decreased in hHTg rats when compared to control rats (P<0.001), but was not affected by insulin. In white adipose tissue (WAT), both the total (P<0.05) and IRS-1 associated PI 3-kinase activities (P<0.001) were increased after 90 min hyperinsulinemia in control animals but not in hHTg animals. CONCLUSIONS: Long-term activation of PI 3-kinase activity by insulin in vivo involves IRS-1 in white adipose tissue, but not in skeletal muscle which implies tissue specificity. The impairment in the PI 3-kinase activation by insulin in hHTg rats may participate in insulin resistance of these animals. PMID- 10467425 TI - Distribution of calcium sensing receptor in rats: an immunohistochemical study. AB - OBJECTIVE: To investigate the organ distribution of calcium sensing receptor (CaR) in rats by immunohistochemical method. METHODS: CaR was identified immunohistochemically in the rat tissues using specific anti-CaR antiserum raised in New Zealand white rabbits immunized with a conjugate of synthetic CaR peptide (186-204) with bovine serum albumin. Immunohistochemical analysis was performed by avidin-biotin complex method. RESULTS: CaR immunoreactivity was visualized in the central nervous system, anterior pituitary, gastric mucosa, small intestine and colon, Auerbach,s and Meissner,s gastric nervous branch, small intestine and colon, pancreas, adrenal medulla, kidney and testis. When using antiserum preincubated with synthetic CaR peptide (186-206) or kidney homogenates, no significant stain of kidney was detected. CONCLUSIONS: The findings suggest that CaR is widely distributed and that the method used is valuable in studying the distribution of caR in rat. PMID- 10467426 TI - Postnatal monosodium glutamate treatment results in attenuation of corticosterone metabolic rate in adult rats. AB - OBJECTIVE: To study the basal and ACTH stimulated production of corticosterone by adrenal cortex on one hand and the binding and degradation of corticosterone in the liver of adult rats which were treated with monosodium glutamate (MSG) during the neonatal period. METHODS: Male offsprings of Sprague-Dawley rats were injected i.p. with MSG (4 mg/g of b.w. in saline) on alternated days for the first 10 days of life, their littermates being used as controls. On the 21st postnatal day they were weaned and used for the observation at the age of 65-75 days. After sacrifice the level of corticosterone in serum and the release of corticosterone from incubated adrenals under basal and ACTH stimulated conditions (ACTH in 6 concentrations from 1.25 to 80 mU/ml medium) were estimated. In addition, glucocorticoid binding to cytosol receptors in the liver and muscle tissues was determined. Corticosterone degradation rate was measured by decrease of corticosterone concentration added to the medium after the incubation with liver slices. RESULTS: Adult rats neonatally treated with MSG had reduced weight of adrenal glands, while plasma corticosterone levels and its basal production by adrenals in vitro were significantly higher than in controls. In MSG treated rats the stimulation of corticosterone production by ACTH was diminished. Glucocorticoid binding to liver cytosolic receptors was significantly decreased, while that in muscle tissue was only slightly elevated. Moreover, a decreased corticosterone degradation rate in liver slices was observed in rats treated neonatally with MSG. CONCLUSIONS: These results are in agreement with previously observed decrease of corticosterone clearance rate in MSG treated animals and suggest that elevated corticosterone levels in plasma and its prolonged response to stressogenic stimulation are due to elevated corticosterone production in adrenals and lower degradation rate in the liver. PMID- 10467427 TI - Pineal gland of the Kuwaiti desert gerbil (Gerbillus cheesmani): alterations of its structure by bromocriptine treatment. AB - In the present study, the pineal gland of the gerbil Gerbillus cheesmani was described for the first time. According to their electron density, two distinct cell types were observed (light and dark pinealocytes). The nuclei were either oval or irregular. Moderate amount of granular endoplasmic reticulum (GER) was observed. Free ribosomes were present throughout the cytoplasm. Mitochondria and lysosomes were among the most common organelles in the pinealocytes. Several dense core vesicles (DCV) were also noted. Blood capillaries with nonfenestrated endothelium were frequent. Bromocriptine treatment for two weeks influenced, to a degree, the physiology of the pinealocytes. It induced a loss of distinction between light and dark pinealocytes, a decrease in the amount of GER and a reduced frequency of lysosomes. On the contrary, lipid droplets and membrane bound vesicles became more frequent. PMID- 10467428 TI - Chromogranin A-, somatostatin- and serotonin- containing endocrine cells in the corporal gastric mucosa of patients with Helicobacter pylori associated chronic gastritis. AB - OBJECTIVE: To examine the number and distribution of chromogranin A, somatostatin and serotonin containing endocrine cells (EC) in the corporal gastric mucosa of patients with Helicobacter pylori (HP) associated chronic gastritis. METHODS: In 12 patients (7 males and 5 females, median age 47 years, range 39-59 years) the number of chromogranin A, somatostatin and serotonin containing endocrine cells (EC) in the corporal gastric mucosa was counted after the appropriate histochemical reaction using specific primary antibodies. The gastric mucosal tissue was obtained by endoscopic biopsy from the greater curvature, anterior and posterior walls of the stomach body. The number of EC was determined per 1 mm2 of the lamina muscularis mucosae. RESULTS: The number of chromogranin A, somatostatin and serotonin containing EC in the corporal gastric mucosa in the patients examined was not increased. CONCLUSIONS: The lack of changes in the somatostatin containing cells indicates that they are not directly involved in the development of hypergastrinaemia accompanying the HP infection. PMID- 10467429 TI - Rapid transient increase of biliary triiodothyronine excretion during short-term infusion of glucose and arginine in rats. AB - OBJECTIVE: To estimate the excretion of triiodothyronine by bile in groups of rats infused with glucose, arginine or glucose combined with various doses of insulin. MATERIALS AND METHODS: Groups of about eight male Wistar Olac rats were anesthetized by pentobarbital and thin polyethylene tubings were inserted into bile duct and femoral vein. The bile was collected into pre-weighed glass vials which were changed every hour. The first one hour interval was considered as control and during the second hour the following i.v. infusions were applied: 1. 2.4 ml 30 % glucose; 2. arginine (80 mg/2. 4 ml saline); 3. 2.4 ml 30 % glucose containing 62.5, 125, 250 or 500 mU insulin. In some groups cycloheximide (2.5 mg/kg) or somatostatin (20 microg/kg) were used. The aliquots of bile were treated with beta-glucuronidase/arylsulfatase and the concentration of total (i.e. conjugated plus unconjugated) triiodothyronine was estimated by specific inhouse radioimmunoassay. The results were expressed as ng/hr and the volume of bile was estimated by weighing the previously tared collection vials. RESULTS: Significant increase of biliary T3 excretion was found during the 60 min infusion of glucose or arginine. However, in fed rats such increase did not continue after the termination of infusion, while in fasted rats the increase was observed still for next 60 min after the infusion. The attempts to further stimulate the excretion of T3 by the addition of small insulin doses (62.5, 125, 250 and 500 mU) to the infused glucose showed inversed effect: by such intervention the increase of T3 was blunted by higher doses, while at lower doses unsignificant increase appeared. The increase of biliary T3 excretion was also blunted by cycloheximide (translation inhibitor) and somatostatin (insulin release inhibitor) both in normal fed and 24 hr fasted rats. CONCLUSIONS: Short-term i.v. infusion of glucose and arginine resulted in immediate and transient increase of biliary T3 excretion which was inhibited by both the cycloheximide and somatostatin. Taken together with our previous findings, this supports the view on rapid fluctuation of hepatic iodothyronine metabolism as related to preprandial (prevailing effect of gluconeogenetic hormones resulting in preferential formation of rT3) and postprandial period (predominant effect of insulin resulting in preferential formation of T3). PMID- 10467430 TI - Insulin-like growth factor binding proteins and their functions (minireview). AB - Insulin-like growth factor (IGF) action is influenced by the insulin-like growth factor binding proteins (IGFBPs). Since 1988 eight forms of IGFBPs have been found which differ in molecular weight, amino acid composition, distribution in biological fluids and influence upon IGF activity. An important biological property of the IGFBPs is their ability to increase the halflife of the IGFs in the blood. They are able to act as potentiators of IGFs activity on the cell proliferation. As IGFBPs bind to cell surfaces, they may act either to deliver the IGFs to those surfaces either for the activation of specific receptors or cell responses independently of receptor activation. Posttranslation modification such as phosphorylation, glycosylation and proteolysis of IGFBPs influence their affinity to IGFs. In addition, the IGFBPs may also act as inhibitors to block the activity of the IGFs by preventing cellsurface binding. PMID- 10467431 TI - MAMMARY-SPECIFIC TRANSGENES AS TOOLS IN SCIENCE AND INDUSTRY (Invited Paper). AB - The ability to modify the animal genome by microinjection of DNA fragments into fertilized embryos was first achieved in 1980. The most abundant mammary-specific milk protein genes were recognized shortly thereafter as valuable tools with which to apply this technology to the mammary gland. The expression of the six major milk protein genes accounts for up to 80 % of the total mRNA in lactating mammary tissue. A few of these genes function efficiently as transgenes and can target the expression of heterologous gene product in a tissue-specific, developmentally regulated fashion. The use of regulatory DNA from these genes in a variety of transgene configurations has resulted in scientific advances in three areas. First, a better understanding of milk protein gene expression has enabled the development of helpful guidelines for the construction of efficiently expressed transgenes, including a requirement for regulatory elements within flanking DNA, the presence of introns, processing signals, and translation and protein processing signals. Second, the ability of the mammary gland to exhibit efficient production of heterologous gene products has been tested and the limitations identified. Third, the ability to produce defined, tissue-specific genetic modifications within the mammary gland has allowed researchers to drastically change its development. Such changes could lead to alterations in the physical or chemical properties of manufactured milk, alterations in milk yield or milk composition, alterations in the metabolism and disease resistance of the lactating female, and alterations in the growth and development of the suckling neonate. This review summarizes the current state of transgenic technology as it relates to mammary specific transgenes and illustrates the application of these transgenes to scientific endeavors and to areas of potential commercial interest. PMID- 10467432 TI - DISTRIBUTION OF SOMATOSTATIN RECEPTOR TYPE 3 IN THE RAT: IMMUNOHISTOCHEMICAL STUDY. AB - Somatostatin receptor type 3 (SSTR-3) was identified immunohistochemically in the rat tissues using specific anti-SSTR-3 serum which was raised in New Zealand white rabbits immunized with a conjugate of synthetic SSTR-3 peptide (28-41) with bovine serum albumin. Immunohistochemical analysis was performed by avidin-biotin complex method. SSTR-3 immunoreactivity was visualized in the central nervous system, anterior pituitary, gastric and duodenal mucosa, Auerbach's and Meissner's nervous branch of gastrointestinal tract, adrenal medulla, testis and pancreas. Significant staining was detected in neural perikarya, axons and dendrites. When using antiserum preincubated with synthetic SSTR-3 peptide (28 41) or rat anterior pituitary homogenate which contains SSTR-3 peptide, no significant stain of the anterior pituitary or neurons in the hypothalamus was detected. These findings suggest that SSTR-3 is widely distributed and that this method is valuable in studying the distribution of SSTR-3 in rats. PMID- 10467433 TI - ASSESSMENT OF BETA-ENDORPHIN AND MELATONIN CIRCADIAN RHYTHMS IN NORMOTENSIVE OBESE WOMEN OF REPRODUCTIVE AGE. AB - We have evaluated the relationship between beta-endorphin (beta-EP) and melatonin (MEL) secretion as determined at 3 h intervals over a 24 h period in 27 normotensive obese women of different phenotypes (12 showing gynoid-type and 15 showing android-type of adipose tissue distribution) as well as in 12 healthy volunteers with normal body weight, aged 30 to 40 years. A considerable increase of mean 24 h beta-EP secretion (mainly due to elevated afternoon and evening levels) and total absence of beta-EP circadian rhythm were observed in all obese patients. Mean 24 h MEL concentrations were markedly higher (mainly due to increased daytime levels) in all obese patients while the disturbances of MEL secretion in the form of acrophase shift and (or) suppression of its rhythmicity were observed especially in obese women with android phenotype. Circadian beta-EP levels correlated positively with BMI and WHR values and negatively with circadian MEL concentrations. In contrast, no significant correlation was found between the values of BMI, WHR and MEL levels. Our data indicate that alterations of beta-EP secretion occurring in obese women could play a role in inducing disturbances of MEL secretory pattern. PMID- 10467434 TI - INFLUENCE OF EGF ON THE RELATIVE VOLUMES OF COLLOID, EPITHELIUM AND STROMA AND ON THYROCYTE NUCLEI SIZE IN RAT THYROID LOBES IN VITRO. AB - The epidermal growth factor (EGF) is believed to be a potent growth factor for the thyroid gland. In the present study, we have examined the relative volumes of the main histological compartments (colloid, epithelium and stroma) and the size of thyrocyte nuclei (the mean volume, the mean intersection area and the mean perimeter) in the rat thyroid lobes incubated in vitro for 18 hrs with EGF, applied in 5 different concentrations: 0.1, 1.0, 10, 100 and 1000 ng/ml. Morphometric evaluation was performed, using a computer image analysis system, developed by us. We found that EGF - in concentration of 100 ng/ml - increased the relative volume of stroma when compared to controls, as well as to all the other groups incubated in exposure to that growth factor (used in different concentrations); at the same time, EGF decreased the relative volume of epithelium in the thyroid gland (statistical significance has been recorded only vs. EGF concentrations of 10 ng/ml and 1000 ng/ml). On the other hand, we observed that EGF - in concentration of 100 ng/ml - significantly increased the mean nuclear volume and the mean intersection area of thyrocyte nuclei when compared to the controls, as well as to EGF in concentrations of 1 ng/ml and 1000 ng/ml. With regards to the mean perimeter, a significant increase of its length was noted in the EGF(100 ng/ml)-exposed group vs. the group incubated with an addition of EGF (1 ng/ml). PMID- 10467435 TI - CAFFEINE INJECTION IN THE DARK PHASE PROLONGS THE NOCTURNAL RISE IN SEROTONIN N ACETYLTRANSFERASE ACTIVITY AND MELATONIN CONTENT IN THE PINEAL GLAND OF MALE RATS. AB - Caffeine, an important member of methylxanthines, induced a prolonged nocturnal rise in pineal melatonin content and an increase in its rate-limiting enzyme serotonin N-acetyltransferase (NAT) activity. The highest levels were reached five hours after subcutaneous caffeine injection to male rats in the dark phase, where the NAT activity increased from 920+/-70 pM/pineal/h in the control group to 1190+/-120 pM/pineal/h (P<0.001) in the treated group. The pineal melatonin content, as well, was elevated from 520+/-40 pg/pineal in the control group to 1120+/-80 pg/pineal (P<0.001) in caffeine treated group. These changes could be attributed to the depressive effect of caffeine on the activity of phosphodiesterase (PDE), the enzyme responsible for the hydrolysis of the intracellular second messenger cyclic adenosine monophosphate (cAMP). PMID- 10467436 TI - MINIREVIEW 7-HYDROXYLATED DERIVATIVES OF DEHYDROEPIANDROSTERONE: WHAT ARE THEY GOOD FOR? AB - In the late fifties and early sixties our group has found that 3beta,7alpha dihydroxy-5-androsten-17-one (7alpha-hydroxy-dehydroepiandrosterone) is a natural constituent of human body fluids. Later, the enzyme activity responsible for its formation has been demonstrated in many animal and human tissues including the foetal ones. The physiological role of this and related steroids has not been understood well for decades. As late as in 1994 Morfin and his group have shown that 7alpha-hydroxy-dehydroepiandrosterone may be a locally active metabolite, responsible for recently discovered immunostimulatory or immunomodulatory effects of dehydroepiandrosterone. PMID- 10467437 TI - Splenic infarction from factor V Leiden mutation. PMID- 10467438 TI - [Rapid direct inhibition of thrombocyte receptors: Epifibatid additive in acute coronary syndromes. 65th Annual Meeting of the German Society for Cardiology and Cardiovascular Research. Mannheim, 8 April 1999]. PMID- 10467439 TI - [Improved tumor diagnosis using MRI with organ-specific contrast media. Wiesbaden, 12 May 1999]. PMID- 10467440 TI - [22nd International Congress of the Association of French-speaking Dermatologists. Beyrouth, Lebanon, 1-4 July 1999. Abstracts]. PMID- 10467441 TI - 26th Meeting of the Federation of European Biochemical Societies (FEBS). Nice, France, June 19-24, 1999. Abstracts. PMID- 10467442 TI - XXVIII National Congress of the Italian Society of Endocrinology. Torino, Italy, May 4-8, 1999. Abstracts. PMID- 10467443 TI - Oral health policies. American Academy of Pediatric Dentistry. PMID- 10467444 TI - Guidelines. American Academy of Pediatric Dentistry. PMID- 10467445 TI - Quality assurance criteria. American Academy of Pediatric Dentistry. PMID- 10467446 TI - Aging in Europe--European Aging. IVth European Congress of Gerontology. Berlin, Germany, July 7-11, 1999. Abstracts. PMID- 10467447 TI - Applied studies of the plant meridian system: II. Agri-wave technology increases the yield and quality of spinach and lettuce and enhances the disease resistant properties of spinach. AB - Agri-wave technology is composed of both a special frequency sound wave and a microelement fertilizer. In both components, the effect of sound waves on plants is more than that of fertilizers, but the best function is a combination of the two. Treatment by Agri-wave technology stimulated the growth rate and increased the yield of spinach. In small plot tests, the length and width of the treated spinach leaf was 50.8 cm and 20.3 cm, respectively, whereas the untreated leaves were 29.20 cm and 8.9 cm. The fresh weight of treated spinach was 0.42 kg. This was 5.5 times higher than that of the untreated spinach. In large area testing (17 hectares), the results of two tests show that the yields of the treated spinach were increased 22.7% and 22.2% over those of the control group. Sugar content of the treated spinach was increased by 37.5%, vitamin A, C, and B were increased 35.63%, 41.67% and 40.00%, respectively, above the levels of the control group. Niacin content was decreased by 7.69%. Of 33 elements analyzed in the spinach, 29 elements were increased by Agri-wave technology. The spinach was infected with "rot disease" in the control group while there was no disease present in the treated group. In greenhouse testing, the average weight of 3 species of lettuce treated by Agri-wave technology was increased 44.10% over that of the control group (P < 0.0001). The average weight of 3 species of lettuce by only sound and only fertilizer treated separately increased 29.92% and 16.19% above that of the control group (P < 0.0001). Sampling survey results in the field test were comparable to the above mentioned greenhouse test. The fresh weight of treated lettuce by Agri-wave technology was increased 41.67% over that of the control group (P < 0.0001). The fresh weight of treated lettuce by only sound and only fertilizer was increased 30.88% and 19.61%, respectively, over the control group (both P < 0.0001). PMID- 10467448 TI - Effects of acupuncture and an agonist of opiate receptors on heroin dependent patients. AB - This study examined the influence of acupuncture and an agonist of opiate receptors (Procedol) by means of temperature in both the left and right auricular "Lung" points, and on the coefficient of interhemisphere asymmetry. Twenty heroin dependent patients participated in this research. They were evenly divided into two groups. Both groups consisted of 10 male drug addicts (18-19 years old) who had been using heroin for 8-10 months. Group 1 received 1.0 ml procedol (i.m.). Group 2 were treated with acupuncture. Temperature was measured on the acupuncture points before and after treatment. Results of this study showed that temperature increased on the left and decreased on the right acupuncture points in both the procedol and acupuncture treated groups. The net changes before and after treatment were 92.87% and 92.53% in groups 1 and 2 respectively. The therapeutic effect of acupuncture on treatment of heroin dependence is through the same pathway as the agonist on opiate receptors, 20% of patients are resisted to acupuncture. PMID- 10467449 TI - Effect of acupuncture at nei-kuan on left ventricular function in patients with coronary artery disease. AB - Effect of acupuncture at Nei-Kuan (EH-6) on left ventricular ejection fraction (LVEF) was examined in 22 patients with angiographically proved coronary artery disease (CAD) and 22 normal subjects. Serial equilibrium radionuclide angiography was done to measure LVEF at 4 different times (at baseline, at 1 to 15 minutes, and 16 to 30 minutes during acupuncture, and immediately after acupuncture). One week later, each patient had an identical imaging protocol with acupuncture performed at a dummy point. Our results showed that in normal subjects, the mean values of LVEF did not change significantly during or after acupuncture. In contrast, in patients with CAD, the mean values of LVEF in the initial 15 minutes of acupuncture significantly increased from baseline (42.5 +/- 15.6% vs. 40.6 +/- 15.4%, p < 0.05). The increase persisted through the next 15 minutes of acupuncture and 15 minutes after acupuncture, but became insignificant at one week. Thus, acupuncture at Nei-Kuan can temporarily improve LV function in patients with CAD. PMID- 10467450 TI - Therapeutic effects of moxibustion on experimental tumor. AB - This work investigated the therapeutic effects of the improved form of moxibustion (MT) on experimental tumor. Sarcoma 180 cells (1 x 10(7)) were transplanted into the subcutaneous tissue in the breast area of female ICR mice. Mice bearing a tumor were divided into one control and four experimental groups. The experimental groups were treated with MT for 1, 2, 3 and 4 times (abbreviated as MT1, MT2, MT3, MT4, respectively). This study showed that the experimental group treated with MT3 displayed the optimal therapeutic response. The longest mean survival time (87.8 days) within 120 days after treatment of MT3 significantly differed from the control group (60.2 days). In addition, uptake of 86Rb-radioactive tracer significantly decreased in tumors treated with MT3. The improved form of moxibustion used in this study is a reliable model of localized hyperthermia in tumor therapy. PMID- 10467451 TI - Effects of ChunDoSunBup Qi-training on growth hormone, insulin-like growth factor I, and testosterone in young and elderly subjects. AB - We observed the response of plasma growth hormone (GH), insulin-like growth factor-I (IGF-I) and testosterone (T) to an acute period of ChunDoSunBup (CDSB) Qi-training. Although the basal level of GH was not different between the two groups, after the portion of the training in which the subjects were physically active (the mid-training point), plasma GH levels increased by 7.26 fold (p < 0.05) in the elderly trainees and by 1.66 fold (p < 0.05) in the young. In response to CDSB Qi-training, IGF-I levels in the young increased significantly at mid-training point, but there were no increase in the elderly. Significant correlations existed between GH and IGF-I levels in the young subjects, but not in the elderly. The T level at the mid-training point increased significantly in elderly subjects but not in the young. These results suggest that CDSB Qi training is a potential method for modulating of the secretion of growth factor in the young and the elderly, but that the elderly IGF-I response does not equal that of the young. In addition, our study suggests CDSB Qi-training may be one mode of therapy applicable to growth factor related disorders such as GH deficiency in children and osteoporosis in the elderly, especially women. PMID- 10467452 TI - Studies on the psychosomatic functioning of ill-health according to eastern and Western medicine. 2. Anxiety-affinitive constitution associated with qi, blood, and body fluid--diagnostic and therapeutic methods. AB - The objective of this investigation was to identify characteristics of psychosomatic function in medical students (N = 62, mean age, 23.3, SD, 1.7 years) whose ill-health was related to unbalanced qi, blood, and body fluid, and to develop a diagnosis and treatment method for these conditions. Our study revealed complicated characteristics. At the psychological level, these characteristics are stress-related emotional disturbances, including anxiety, insomnia and anergy, and the lowering of social function. At the physiological level, these characteristics are associated with high complexity of fractal dimension of eye (horizontal) and respiratory (thoracic) movements correlated to STA1-trait anxiety. Thus, the three psychosomatic characteristics related to unbalanced qi, blood, and body fluid suggest the concept of an anxiety-affinitive constitution, also described as the equivalent of ill-health. This indicates that diagnosis and treatment of this type of constitution has the potential to be useful for both the prevention of stress-related and life-style disease, and the treatment of current disease. PMID- 10467453 TI - Juzen-taiho-to, a Japanese herbal medicine, modulates type 1 and type 2 T cell responses in old BALB/c mice. AB - Aging is associated with skewed type 2 (T2) T cell responses that may be modulated by herbal medicines. A group of Japanese herbal medicines, so-called "Hozai," have been used to improve the physical condition of the elderly. One representative "Hozai," Juzen-Taiho-To (JTX) appears to have beneficial effects on cancer patients. In this study we hypothesized that JTX modulated skewed T2 responses in the elderly. T1 and T2 responses against ovalbumin (OVA) were examined in old BALB/c mice fed JTX (0.2% w/w). We measured anti-OVA IgG1, IgG2a, and IgG2b antibody (Ab) levels after the primary and secondary OVA challenges; T1 and T2 responses augment IgG2a/IgG2b Ab and IgG1/IgE Ab production, respectively. We also assessed production of T1 and T2 cytokines (IFN-gamma and IL-5, respectively), and co-stimulatory molecule expression by regional draining lymph node cells. JTX-fed mice had higher IgG2b Ab and IFN-gamma production than controls along with lower IgG1 Ab. JTX did not alter IL-5 production or co stimulatory molecule expression. Hoelen, an herbal component, induced similar changes. Our results indicate that JTX and Hoelen modulate T cell responses against OVA toward more balanced T1/T2 responses in old BALB/c mice. Such effects of JTX may help prevent the development of diseases associated with immunodisregulation in the elderly. PMID- 10467454 TI - The effect of da-fang-feng-tang on treatment of type II collagen-induced arthritis in DBA/1 mice. AB - Da-Fang-Feng-Tang (DFFT), which is believed to be effective for treating human rheumatoid arthritis (RA), was given to DBA/1 mice at the onset of type II collagen-induced arthritis (CIA) to examine its effect. Granules of the crude DFFT extract were administered by gastric gavage at a dose of 1.6 g/kg/day for 12 weeks, starting the day CIA began. The levels of anticollagen IgG antibody were significantly decreased in the sera of the DFFT-treated group compared with the control group from weeks 2 to 7 after the onset of CIA. The severity of arthritis in the DFFT-treated group was markedly alleviated when compared with the control group. In addition, histological examination of the DFFT-treated group showed less cartilage and bone erosion. These results suggest that administration of DFFT suppressed the development of CIA in mice and support the belief that DFFT is effective in treating human RA. PMID- 10467455 TI - Induction of T lymphocyte apoptosis by treatment with glycyrrhizin. AB - The effect of glycyrrhizin (GL), a Chinese herbal drug extracted from licolice roots, on murine lymphocytes for inducing apoptotic cell death was studied. Addition of GL (25-400 micrograms/ml) to cultured splenocytes and thymocytes from BALB/c mice definitely promoted DNA fragmentation. A single injection of GL (100 micrograms/mouse) into BALB/c mice did not cause any detectable DNA fragmentation or cell death of splenocytes and thymocytes. Cytofluorometric analysis of these cells, however, demonstrated a reduction in mitochondrial transmembrane potentials (delta psi m). Repeated injections of GL (100 micrograms/mouse/day) into mice for 7 days actually resulted in induction of low grade DNA fragmentation selectively in splenocytes. Cell population analysis of viable lymphocytes suggested that both CD4+ Th lymphocytes and CD8+ Tc lymphocytes may have been relatively more sensitive than B220+ B lymphocytes for the apoptotic cell death. We concluded from these results that GL acts as a rather selective inducer of mature T lymphocyte apoptosis with a reduction in delta psi m potentially preceding lymphocyte death. PMID- 10467456 TI - The anti-inflammatory and hepatoprotective effects of fractions from Cudrania cochinchinensis var. gerontogea. AB - Various fractions of the ethanol extract from the root wood of Cudrania cochinchinensis var. gerontogea (Moraceae) were evaluated for their anti inflammatory effects on carrageenan-induced edema and hepatoprotective activities on carbon tetrachloride (CCl4)-induced and D-galactosamine-(D-GalN) induced acute hepatotoxicity in rats. The fractions (n-hexane, CHCl3, EtOAc, n-BuOH, and H2O) displayed significant inhibitory activity against carrageenan-induced edema, and the active anti-inflammatory components were further localized in the n-BuOH fraction, which exhibited the greatest anti-inflammatory effect, an effect 5% greater than indomethacin (which was used as a standard reference substance). Each fraction exerted a significant hepatoprotective effect by reducing enzymatic alteration (sGOT and sGPT) and by improving hepatic lesions, including liver centrilobular inflammation, cell necrosis, fatty change, ballooning degeneration in CCl4-induced acute hepatitis; and necrosis of the portal area in D-GalN induced acute liver injury. The n-BuOH and EtOAc fractions had the greatest hepatoprotective effects on CCl4-induced liver injury; in contrast, the CHCl3 fraction was most potent against D-GalN intoxication, which is comparable to silymarin, as a recognized hepatoprotective drug. PMID- 10467457 TI - Effect of a urinary preparation on liver injury by short-term carbon tetrachloride treatment in rats. AB - The hepatoprotective effect of a preparation of human urine (PHU) was assessed against short-term carbon tetrachloride (CCl4) treatment in rats. Significant prevention of liver injury by PHU was found after CCl4 treatment, judging by the changes of serum biochemical parameters, and hepatic protein and triglyceride contents. The increased liver lipid peroxidation, and decreased liver vitamin C concentrations observed after CCl4 treatment were significantly prevented by PHU administration. The increase in liver glutathione (GSH) contents observed after CCl4 treatment was further increased by PHU treatment. Liver catalase activity decreased after CCl4 treatment, while liver superoxide dismutase and GSH peroxidase activities did not change. PHU administration further inhibited the decrease in liver catalase activity after CCl4 treatment. These results indicate that PHU administration can prevent liver injury induced by CCl4 in rats by inhibiting enhanced lipid peroxidation and by improving disrupted active oxygen metabolism in the injured liver. PMID- 10467458 TI - Orally administered Panax ginseng extract decreases platelet adhesiveness in 66% hepatectomized rats. AB - The effect of oral administration of Panax ginseng extract (GE) on platelet adhesiveness was examined in 66% hepatectomized rats. A significant decrease in platelet adhesiveness was obtained when 125 mg/kg/day GE was administered for 6 days before and after hepatectomy. The total cholesterol concentration in the serum was also decreased by GE administration. Food intake was unaffected by GE administration. Serum parameters indicating liver and kidney function were unchanged after GE administration except for lipid metabolic parameters. Because enhanced platelet adhesiveness and hyperlipidemia induces atherosclerosis, these results suggest that orally administered GE is capable of improving the atherosclerotic condition associated with hepatectomy. PMID- 10467459 TI - Anticonvulsant effect of Uncaria rhynchophylla (Miq) Jack. in rats with kainic acid-induced epileptic seizure. AB - This study investigated the anticonvulsant effect of Uncaria rhynchophylla (UR) and the physiological mechanisms of its action in rats. A total of 70 male Sprague-Dawley (SD) rats were selected for study. Thirty four of these rats were divided into 5 groups as follows: 1) CONTROL GROUP (n = 6): received intraperitoneal injection (i.p.) of kainic acid (KA, 12 mg/kg); 2) UR1000 group (n = 10), 3) UR500 group (n = 6) 4) UR250 group, received UR 1000, 500, 250 mg/kg i.p. 30 min prior to KA administration, respectively; 5) Contrast group: received carbamazepine 20 mg/kg i.p. 30 min prior to KA administration. Behavior and EEG were monitored from 15 min prior to drug administration to 3 hours after KA administration. The number of wet dog shakes were counted at 10 min intervals throughout the experimental course. The remaining 36 rats were used to measure the lipid peroxide level in the cerebral cortex one hour after KA administration. These rats were divided into 6 groups of 6 rats as follows: 1) Normal group: no treatment was given; 2) CONTROL GROUP: received KA (12 mg/kg) i.p.; 3) UR1000 group, 4) UR500 group, 5) UR250 group, received UR 1000, 500, 250 mg/kg i.p. 30 min prior to KA administration, respectively; 6) Contrast group: received carbamazepine 20 mg/kg i.p. 30 min prior to KA administration. Our results indicated that both UR 1000 and 500 mg/kg decreased the incidence of KA-induced wet dog shakes, no similar effect was observed in the UR 250 mg/kg and carbamazepine 20 mg/kg group. Treatment with UR 1000 mg/kg, 500 mg/kg, or 250 mg/kg and carbamazepine 20 mg/kg decreased KA-induced lipid peroxide level in the cerebral cortex and was dose-dependent. These findings suggest that the anticonvulsant effect of UR possibly results from its suppressive effect on lipid peroxidation in the brain. PMID- 10467460 TI - Effects of berberine on arylamine N-acetyltransferase activity in human colon tumor cells. AB - Berberine was used to determine loss of viable cells and inhibition of arylamine Nacetyltransferase (NAT) activity in a human colon tumor (adenocarcinoma) cell line. The viable cells were determined by trypan blue exclusion under a light microscope. The NAT activity was measured by high performance liquid chromatography for the amounts of N-acetyl-2-aminofluorene (AAF), N-acetyl-p aminobenzoic acid (N-Ac-PABA), and the remaining 2-aminofluorene (AF) and p aminobenzoic acid (PABA). The viability and NAT activity in a human colon tumor cell line was inhibited by berberine in a dose-dependent manner, i.e., the higher the concentration of berberine, the higher the inhibition of NAT activity and cell death. The NAT activities measured in the intact human colon tumor cells were decreased over 50% by AAF and NAc-PABA production from acetylation of AF and PABA. The apparent values of Kmoff and Vmax of NAT from colon tumor cells were also inhibited by berberine in cytosols and in intact cells. This report is the first to show that berberine did affect human colon tumor cell NAT activity. PMID- 10467461 TI - Miscarriage prevention tea affects plasma beta-endorphin concentrations in women with early threatened abortions. AB - Twenty threatened abortive patients in the 7-8th week of gestation were treated with a classical miscarriage prevention tea (Shou-Tai-Tang) combined with psychological consultation. All of the patients had a history of unexplained recurrent abortions. This treatment succeeded in sixteen out of 20 patients. The plasma concentrations of beta-endorphin (beta-EP), gonadotrophin releasing hormone (GnRH), human chorionic gonadotrophin (hCG), and progesterone (P4) were measured by radioimmunoassay before and after treatment. Compared to control subjects, beta-EP levels were significantly higher, while GnRH, hCG, and P4 were lower than before treatment. Concentrations of these peptides/hormones returned to normal ranges after successful treatment. PMID- 10467462 TI - Effect of Koso-san on globus pharyngeus. AB - Twenty-three patients with globus pharyngeus were treated with Koso-san (TJ-70) at a dose of 7.5 g/day for at least 14 days. Symptoms disappeared in 18 cases and improved in 3 cases, therefore, the effective rate was 91.3% (21/23). The symptoms disappeared on average within 13.5 days. Terasawa qi-stasis scores were significantly decreased after the treatment. TJ-70 might thus be a remedy for globus pharyngeus with qi-stasis. PMID- 10467463 TI - International viewpoints. PMID- 10467465 TI - The controversy of significance testing: misconceptions and alternatives. AB - The current debate about the merits of null hypothesis significance testing, even though provocative, is not particularly novel. The significance testing approach has had defenders and opponents for decades, especially within the social sciences, where reliance on the use of significance testing has historically been heavy. The primary concerns have been (1) the misuse of significance testing, (2) the misinterpretation of P values, and (3) the lack of accompanying statistics, such as effect sizes and confidence intervals, that would provide a broader picture into the researcher's data analysis and interpretation. This article presents the current thinking, both in favor and against, on significance testing, the virtually unanimous support for reporting effect sizes alongside P values, and the overall implications for practice and application. PMID- 10467464 TI - Evolving trends in critical care nursing practice: results of a certification role delineation study. AB - BACKGROUND: In 1997, the AACN Certification Corporation, in conjunction with Professional Examination Service, undertook a role delineation study as 1 component of a large-scale, comprehensive, and systematic study of practice to update previous data. Focus groups made up of practicing critical care nurses were used to determine trends and changes in adult, pediatric, and neonatal critical care nursing practice. METHODS: Sixteen focus groups (6 adult, 5 pediatric, and 5 neonatal) used specially prepared protocols to guide discussions. Questions were designed to elicit descriptions of changes in critical care nursing practice in the preceding 5 years. Qualitative comments of the participants were analyzed across all the focus groups, rather than separately for the adult, pediatric, and neonatal focus groups. Then data for the focus groups for each patient-age range were aggregated and reviewed to abstract themes. RESULTS: Trends and changes in practice for adult, pediatric, and neonatal critical care nurses were determined. Common themes include ethical and legal issues, changes in the population of patients, psychosocial factors, and the impact of managed care. CONCLUSIONS: The results of these focus groups can be used to update the test blueprints that underlie the CCRN certification examination programs for adult, pediatric, and neonatal critical care nurses. Critical care nursing practice is changing. Specific knowledge of the changes is important for educators, managers, and clinicians. The results of this role delineation study can be used to teach, adapt systems, and validate practice. PMID- 10467466 TI - Card sort: an assessment tool for the educational needs of patients with heart failure. AB - BACKGROUND: Effective education of patients is an integral component in reducing hospital readmissions for patients with heart failure and in improving patients' quality of life. OBJECTIVES: To describe the use of a card sort method for determining the learning needs of patients with heart failure and the ability of nurses and physicians to accurately predict patients' self-determined needs. METHODS: Thirty patients were asked to sort 12 cards with questions related to the content of discharge teaching for patients with heart failure. The nurse and the physician caring for each patient were instructed to sort the cards independently to characterize the learning needs of the patient. RESULTS: Patients most often selected the following as the most important educational topics: What is wrong with my heart? What is the future of my disease? and How will I know if my heart failure is getting worse? Physicians selected the identical top question as their patients did 17% of the time. Physicians matched any of their patients' top 3 choices 34% of the time. Questions selected as most important by patients were chosen by physicians as least important 13% of the time. Nurses chose the same top question as their patients did 23% of the time. Nurses matched any of their patients' top 3 choices 34% of the time. Nurses selected opposite choices from those of their patients 6% of the time. CONCLUSIONS: The card sort method may be an effective tool to ascertain the individual learning needs of patients with heart failure. PMID- 10467467 TI - Nursing interventions to decrease bleeding at the femoral access site after percutaneous coronary intervention. SANDBAG Nursing Coordinators. Standards of Angioplasty Nursing Techniques to Diminish Bleeding Around the Groin. AB - BACKGROUND: This trial is the first prospective, multicenter clinical nursing trial conducted to measure the effect of nursing interventions on bleeding at the femoral access site after percutaneous coronary intervention with or without a potent antiplatelet agent given along with heparin and aspirin. OBJECTIVE: To measure the relationship between nursing interventions and complications at the arterial access site in patients undergoing percutaneous coronary interventions and to recommend a standard of care to minimize bleeding complications. METHODS: In a descriptive, correlational 4010-patient study, nursing care interventions after coronary procedures were measured. Observed standards of care were assessed, and regression techniques were used to evaluate nursing interventions and the effect of the interventions on bleeding at the access site after percutaneous coronary procedures. RESULTS: Several significant correlations between nursing interventions and the occurrences of moderate to severe bleeding at the access site were found; however, most interventions had little effect. The most significant factors in decreasing complications at the access site were early removal of the arterial sheath, the type of pressure mechanism used to achieve arterial hemostasis, staffing allocation, and the person and method used to remove the sheath. CONCLUSION: Many nursing interventions after percutaneous coronary intervention have become routine in the absence of clinical outcome data. Most nursing interventions aimed at decreasing bleeding at the vascular access site increase nursing workload but do not significantly affect bleeding in the groin. These results underscore the importance of continued clinical research studies to validate nursing practice on the basis of patients' outcomes. PMID- 10467468 TI - Oral care in the adult intensive care unit. AB - BACKGROUND: Nurses have not been formally trained in assessing the oral status of patients in intensive care units, and no oral care protocols for these patients are available. OBJECTIVES: To assess the oral status of patients in an intensive care unit, evaluate the effects of a defined oral care protocol on the oral health status of patients in an intensive care unit, and compare oral assessments of a dental hygienist with those of intensive care nurses. METHODS: A nonequivalent comparison group, longitudinal design with repeated measures was used. In phase 1, oral assessment data on the comparison group were collected by a dental hygienist. In phase 2, nurses were instructed in oral assessment and an oral care protocol. In phase 3, the oral care protocol was implemented in the treatment group, and oral assessment data were collected separately by the dental hygienist and by nurses. RESULTS: The mean inflammation score was significantly lower (t test P = .03) in the treatment group (mean, 3.9; SEM, 3.0) than in the comparison group (mean, 12.4; SEM, 2.2). Although not significant, the mean scores of the treatment group were also lower than those of the comparison group on scales of candidiasis, purulence, bleeding, and plaque. Correlations between scores for individual items on the oral assessment tool obtained by the dental hygienist and those obtained by nurses were all greater than 0.6386. CONCLUSION: Implementation of a well-developed oral care protocol by bedside nurses can improve oral health of patients in the intensive care unit. PMID- 10467469 TI - Effects of position of chest drainage tube on volume drained and pressure. AB - BACKGROUND: The nursing practice of avoiding dependent loops in the tubing of chest drainage systems because such loops may impede drainage and alter the intrapleural pressure is not research based. OBJECTIVES: To determine if the volume of fluid drained and pressure vary when the chest drainage tubing is straight, coiled, has a dependent loop, or has a dependent loop that is periodically lifted and drained. METHODS: A repeated-measures design was used. For each tubing position, 500 mL of fluid was infused into the pleural space of 8 adult pigs during 45 minutes. The volume of fluid drained and the pressure at 2 locations within the drainage tubing were measured for 1 hour. RESULTS: After 60 minutes, significantly less fluid (least significant difference test, P = .03) was drained with the dependent-loop tubing position (65 mL) than with the other 3 positions. However, the amount of fluid drained was not significantly different among the lift and drain (250 mL), coiled (301 mL), or straight (337 mL) tubing positions. Throughout the entire study, pressure at the connection between the chest tube and the drainage tube was significantly higher (least significant difference test, P = .003) for the dependent loop with and without periodic lifting and draining. CONCLUSIONS: Straight and coiled tube positions are optimal for draining fluid from the pleural space. If a dependent loop cannot be avoided, lifting and draining it every 15 minutes will maintain adequate drainage. PMID- 10467470 TI - Mechanical circulatory support as a bridge to cardiac transplantation: toward the 21st century. AB - Mechanical circulatory support is used to sustain the lives of patients awaiting cardiac transplantation who would otherwise die before a donor organ became available. Currently available ventricular assist devices used for mechanical circulatory support, risk factors and complications associated with use of these devices, and selection of candidates for treatment with mechanical support as a bridge to cardiac transplantation are reviewed. The importance of early insertion of the devices before end-organ dysfunction occurs is examined. PMID- 10467471 TI - Life with a left ventricular assist device: the patient's perspective. AB - BACKGROUND: Understanding the psychological experience of living with a life sustaining device is necessary for developing individualized, supportive interventions for patients with a left ventricular assist device. OBJECTIVES: To describe patients' experiences with left ventricular assist devices, strategies for coping with prolonged hospitalization, and suggestions for individualized nursing interventions. METHODS: The phenomenology method of qualitative research was used. Data collection involved unstructured, open-ended interviews of 6 hospitalized subjects who had a pneumatic left ventricular assist device. RESULTS: Patients described coping strategies for prolonged hospitalization, including family support, religious convictions, and diversional activities. Data analysis suggests that critical care nurses can best support these patients by establishing trusting relationships, fostering independence, and incorporating humor into their care. CONCLUSIONS: Patients awaiting cardiac transplantation coped effectively with prolonged hospitalization with a life-sustaining device in place. Spirituality, humor, and strong family relationships contributed to their positive outlook. PMID- 10467472 TI - Reliability of an intravenous intermittent access port (saline lock) for obtaining blood samples for coagulation studies. AB - BACKGROUND: Patients admitted to rule out myocardial infarction often receive a heparin infusion that requires measurement of partial thromboplastin time every 6 to 8 hours until coagulation values stabilize. If the patient's acuity at admission does not necessitate placement of arterial or central catheters, insertion of an intermittent intravenous access port (saline lock) would decrease the need for frequent venipuncture. Many studies on obtaining blood samples via arterial and central catheters have been reported, but no reports have described the method of using a saline lock or the amount of blood discarded during that procedure. OBJECTIVE: To evaluate the efficacy of inserting a saline lock specifically for obtaining blood samples and to determine the amount of blood that must be discarded in order to obtain prothrombin and partial thromboplastin times that match the values from blood samples obtained via venipuncture. METHODS: Coagulation values determined with 2 consecutive blood samples obtained via the saline lock after the first 0.5 mL of blood withdrawn was discarded were compared with measurements determined with a blood sample obtained via venipuncture. RESULTS: Prothrombin and partial thromboplastin times were not significantly different among the 3 blood samples. CONCLUSION: The untoward effects of frequent venipunctures to obtain blood samples for measurement of prothrombin and partial thromboplastin times can be lessened by using an 18-gauge saline lock to obtain blood samples and discarding the first 0.5 mL of blood withdrawn before the samples for coagulation studies are collected. PMID- 10467473 TI - VEGF and TMR: myocardial revascularization in the next millennium. PMID- 10467474 TI - [Training in a simulator. Play or indispensable component of the medical learning process?]. PMID- 10467475 TI - [Simulator training in anesthesia. Applications and value]. AB - Simulation has gained an important role in medical education and continuing education in the field of anaesthesia and emergency medicine. This article gives background information on how full-scale simulators are applied in medical education as well as in continuing education for advanced anesthesia and scientific applications. Acceptance of training seems enhanced by inclusion of the human factor aspect, since this has proven to be a major source for the development of critical situations in our specialty. Furthermore, drawbacks of the simulators available and the current training availability in Germany is described. PMID- 10467476 TI - [Postoperative analgesia in children after hernioplasty. Wound infiltration with different concentrations of bupivacaine: a pilot study]. AB - Wound instillation seems to be an easy and preferable way to achieve postoperative analgesia in pediatric hernioplasty. This prospective, randomized and double-blinded pilot-study was initiated to gain preliminary information in order to define the appropriate concentration of local anaesthetic for efficient posthernioplastic analgesia. METHOD: 29 children aged 3.1 to 13.7 (5.25 (3.8-8.2) years were randomly assigned to receive either 0.2 ml/kg bupivacaine 0.125% (n = 10), bupivacaine 0.25% (n = 10) or bupivacaine 0.5% (n = 9). The local anesthetic (LA) was instillated intraoperatively before wound closure above the external oblique muscle and below Scarpa's fascia. After entering the post-anesthetic care unit (PACU) pain was assessed by a trained nurse using the linear analogue pain scale (LAPS) in intervals of 15 min. Patients were observed in the PACU for 30-60 min. Pain was further evaluated for 5.5(3-6) h in the ward every hour. In day only patients the parents were contacted 24 h postoperatively to obtain additional information. RESULTS: From the beginning of the observation period the 0.5% group tended to have less pain than the others in the PACU. The 0.125% and 0.25% group required earlier supplementary analgetics. In addition, the 0.5% group needed once (1/9) supplementary analgesics; the 0.25% group five times (5/10) and the 0.125% group six times (6/10). None of these results is statistically significant, though they appear to be clinically relevant. DISCUSSION: Wound instillation with 0.2 ml of bupivacaine 0.5% seems to be easy to perform, safe and efficient in controlling posthernioplastic pain. Because of the small numbers of patients included however, no statistically significant differences were observed between the groups. Neither in the LAPS on arrival and observation at the PACU nor in the need for supplementary analgesics. Despite lacking significance the clinical impression suggests a difference to be validated by larger studies. Our data implies that wound instillation with 0.2 ml/kg bupivacaine 0.5% should be regarded for routine usage. PMID- 10467477 TI - [News of ether anesthesia in the local press in the 1847 in Bamburg] 449 -1 [How much blood flow is required by the myocardium?]. AB - The news of success in ether anesthesia on 16 October 1846 in Boston spread like wildfire through Europe. In the German-speaking countries, the national newspapers as conveyers of the development played a prominent role; this was especially the case for the Augsburger Allgemeine Zeitung whose correspondent, Aloys Martin [1], was a medical professional. The function of local newspapers has not yet been investigated. This was to be remedied by examining both of Bamberg's local papers. All issues from 01.01. to 31.04.1847 were surveyed with respect to relevant announcements. These were then catalogued according to date, extent and contents. Starting on 22.01.1847, the events outside of and in Bamberg were reported on in 26 notices and articles of just a few lines to half a page and in advertisements; Aloys Martin also appears on the scene. Thus, the local press in Bamberg also informed its readers early and regularly on the discovery of ether and the succeeding events. PMID- 10467478 TI - [How much blood flow is required by the myocardium?]. AB - OBJECTIVES: The myocardium of the left ventricle exhibits spatial heterogeneity of blood flow under physiological conditions. This study was designed to investigate, whether oxygen supply is jeopardized in low flow areas (blood flow < 50% of mean) under physiological conditions and whether areas of high flow (> 150% of mean) exhibit perfusion in excess of demand ("luxury perfusion"). METHODS: The study was performed in anesthetized and ventilated beagle dogs. Local blood flow was reduced by mechanically narrowing of the r. circumflexus of the left coronary artery; myocardial blood flow was measured by the tracer microsphere technique, free concentrations cellular adenosine by the SAH technique, regional metabolism of substrates by the desoxyglucose-technique. RESULTS: Low flow areas exhibited normal oxygenation of the myocardium, while in high flow areas no luxury perfusion could be demonstrated. CONCLUSION: Myocardial blood flow and metabolism demonstrate significant spatial heterogeneity. There appears to be no absolute threshold of blood flow, where regional myocardial ischemia develops. Probably biochemical evidence of myocardial ischemia is determined by a local ratio of oxygen supply and demand. PMID- 10467479 TI - [Arterial air embolism as a complication of blunt thoracic trauma]. AB - Systemic air embolism is a dramatic event frequently resulting in death. The clinical manifestations are polymorphic and unspecific. We report a case of a systemic air embolism in a patient with lung contusion. We wish to arouse a sensibility for this clinical picture, which seems to be underestimated in its frequency and importance, and discuss concepts of mechanical ventilation in patients with blunt thoracic trauma. PMID- 10467480 TI - [Status epilepticus. Rational diagnosis and current therapeutic concepts]. AB - Status epilepticus (SE) is defined as a seizure persisting over 10 min or repeated seizures without recovery between the attacks. SE is a frequent medical emergency with a high mortality, requiring aggressive and prompt treatment and a systematic, coordinated approach. A variety of new drugs has been introduced for the treatment of SE, including midazolam, propofol, fosphenytoin, and valproate. However, there are very few controlled clinical trials. Benzodiazepines are highly effective in terminating the SE in most patients and should be used as initial treatment. Phenytoin and Phenobarbital are employed if Benzodiazepines are not successful. The optimal therapeutic approach for the patient with refractory SE remains to be defined. Refractory SE has a poor prognosis and should be treated in a specialized ICU with EEG-monitoring. Barbiturates, propofol, and midazolam are substances that can be used alternatively. PMID- 10467481 TI - [History of the development of intensive care medicine. 7. History of continuing education in intensive care]. PMID- 10467482 TI - [Technical developments in the area of anesthesia and anesthesia simulation. A window to the 21st century? Meeting of the Society for Technology in Anesthesia (STA) January, 20-3, 1999, San Diego]. PMID- 10467483 TI - [Is a patient mouth protector really necessary?]. PMID- 10467484 TI - [Tachyphylaxis to local anesthetics]. PMID- 10467485 TI - [Risk in anesthesia in ORL operations in small children]. PMID- 10467486 TI - [Substitution with human albumin]. PMID- 10467487 TI - [Anesthesia in endocrine dysfunction]. PMID- 10467488 TI - Violence in lesbian and gay relationships: theory, prevalence, and correlational factors. AB - This article reviews and critiques the existing empirical literature examining interpersonal violence in lesbian and gay relationships. Studies assessing psychological, physical, and nonconsenting sexual forms of violence in intimate, same-sex relationships are reviewed, and their findings are integrated with what is known about partner abuse in heterosexual relationships. Nineteen studies are described and categorized according to the specific questions being addressed. This body of literature suggests that prevalence rates of same-sex partner abuse are high and its correlates show many similarities to those identified in incidents of heterosexual partner abuse. This article addresses the need for substantially increased efforts in this field of study in terms of well controlled and theory-driven research design. In terms of other implications of this body of literature, the high prevalence rate of partner abuse among lesbian and gay populations needs to be recognized by providers of both physical and mental health services who potentially treat victims, so that they can more accurately identify appropriate interventions. More research is warranted, not only in the general area of lesbian and gay partner abuse, but in examining various treatment modalities and their effectiveness in helping perpetrators to end the cycle of violence. PMID- 10467489 TI - Social anxiety and drinking in college students: a social cognitive theory analysis. AB - Evidence is reviewed that indicates that social anxiety is a significant motivation for drinking among college students. Although the link between social anxiety and alcohol consumption has been studied from a variety of perspectives, there has been little integration of data. Drawing from the alcohol and anxiety literature, the relationship between social anxiety and heavy drinking in college students is examined. Findings indicate a clear relationship between social anxiety and drinking that may be moderated by alcohol expectancies and self efficacy beliefs specific to socially anxious situations. A social cognitive model is proposed to guide future research and intervention efforts. A better understanding of college students' reasons for drinking offers the possibility of improving prevention and treatment efforts designed to reduce excessive drinking. PMID- 10467490 TI - Tourette's and comorbid syndromes: obsessive compulsive and attention deficit hyperactivity disorder. A common etiology? AB - Tourette's syndrome (TS), a neuropsychiatric movement disorder that manifests itself in childhood, is often associated with comorbid symptomatology, such as obsessions, compulsions, hyperactivity, distractibility, and impulsivity. Epidemiological studies suggest that a substantial number of TS patients develop clinical levels of obsessive-compulsive disorder (OCD) and/or attention deficit hyperactivity disorder (ADHD). This review aims to provide an integrated account of the three disorders in terms of their comorbidity. Neuroimaging studies suggest that all three disorders involve neuropathology of the basal-ganglia thalamocortical (BGTC) pathways: TS in the sensorimotor and limbic BGTC circuits; OCD in the prefrontal and limbic BGTC pathways; and ADHD in the sensorimotor, orbitofrontal, and limbic BGTC circuits. The pattern of comorbidity and other evidence indicates that the TS gene(s) may be responsible for a spectrum of disorders, including OCD and ADHD, but also that the disorders OCD and ADHD can exist in their own right with their own etiologies. PMID- 10467491 TI - Psychosocial processes underlying the maintenance and persistence of depression: implications for understanding chronic depression. AB - The three major theoretical perspectives on the maintenance and persistence of depression in the psychological literature are reviewed. Cognitive theorists, such as Teasdale and Nolen-Hoeksema, focus on how a reciprocal relationship between depressed mood and the individual's processing of, and response to, their symptoms maintains and prolongs the duration of depressive episodes. Interpersonal theorists, such as Lewinsohn and Coyne, hypothesize that the depressed person's interpersonal behavior elicits negative reactions from the social environment, leading to a downward spiral of persistent depression. Finally, recent studies have indicated that individuals who experienced childhood adversity are more susceptible to a chronic course of depression. Although these three perspectives vary somewhat in the degree of support they have accumulated, each exhibits some promise for helping to elucidate the maintenance and persistence of depression. However, much of the available empirical literature suffers from a number of critical limitations, including the lack of clinical samples, longitudinal studies, and adequate attention to confounding variables, such as the severity and duration of depression at baseline and comorbid psychopathology. We conclude by suggesting a number of ways in which cognitive and interpersonal factors and early adversity may interact to maintain and prolong depressive episodes. PMID- 10467492 TI - Unconscious processes, subliminal stimulation, and anxiety. AB - Ever since Poetzl's studies, subliminal stimulation has been used as a paradigm to explore the connection between unconscious processes and psychopathology. Inspired by the psychodynamic tradition, folk psychology attributes a dramatic power to subliminal stimulation. In contrast, most modern researchers argue that effects of subliminal stimulation are rather limited. Does that mean that the unconscious is irrelevant to psychopathology? Not necessarily. Ohman and Soares' hypothesis about the preattentive origins of phobic reactions represents a good example of a model in which a "quick and dirty" unconscious may produce pathogenic effects. Although the empirical basis of this model is still meagre, its attractiveness hinges on the assumption that "quick and dirty" processes that make up the first second of human information processing are essential for higher level analysis and performance. In line with this, recent studies have indicated that the attentional bias that accompanies pathological anxiety, might be an unconscious phenomenon. Theories that focus on unconscious cognitive processes involved in pathological anxiety are certainly interesting, but it should be emphasized that there are other aspects of automaticity (i.e., involuntariness) that may be as relevant to psychopathology as absence of awareness. PMID- 10467493 TI - Psychotherapy for male sexual dysfunction: a review. AB - There are nine major diagnostic categories of male sexual dysfunction in the DSM IV. Psychotherapy outcome studies focusing on these problems are reviewed. Currently, there appears to be little evidence that effective psychological interventions exist for these problems. Reasons for this include numerous methodological problems of existing research, effect sizes that are not clearly clinically significant, lack of compelling follow-up data indicating durability of treatment gains, and lack of treatment manuals that allow replication. Also, some disorders have been ignored by researchers. Guidelines for sample selection and description, assessment and diagnosis, and research design are provided. PMID- 10467495 TI - Therapy at its best. PMID- 10467494 TI - Relations among psychological trauma, dissociative phenomena, and trauma-related distress: a review and integration. AB - Recently, there has been a resurgence of interest in relations among psychological trauma, dissociative phenomena, and various forms of trauma-related distress that has spawned a prolific amount of research. To date, a relatively comprehensive review of this recent research is lacking. Thus, this paper provides such a review to help summarize and synthesize recent findings, illuminate study limitations, and offer suggestions for future research. In general, findings have revealed fairly strong and consistent relations among the constructs of trauma, dissociation, and trauma-related distress (e.g., posttraumatic stress disorder, borderline personality disorder, bulimia); individuals who have experienced a traumatic event are more likely to dissociate than individuals who have not, and individuals who experience more dissociative phenomena are more likely to also experience higher levels of trauma-related distress. It is theorized here that dissociative phenomena and subsequent trauma related distress may relate to fears about death and fears about loss or lack of control above and beyond the occurrence of the traumatic event itself. Such fears about death and loss/lack of control may also help differentiate traumatized individuals who psychologically suffer to varying degrees. Possible functions of dissociation in response to trauma and in relation to forms of trauma-related distress are considered and discussed. PMID- 10467496 TI - What's eating you? Centipedes (Chilopoda). PMID- 10467497 TI - Aquatic dermatitis, Part I. Dermatitis caused by coelenterates. PMID- 10467498 TI - Arthropathy associated with cystic acne, hidradenitis suppurativa, and perifolliculitis capitis abscedens et suffodiens: treatment with isotretinoin. AB - A patient with arthropathy associated with cystic acne, hidradenitis suppurativa, and perifolliculitis capitis abscedens et suffodiens who showed a dramatic response to isotretinoin is described. This, to our knowledge, is the first report documenting effective treatment of this condition, whose nosologic position with respect to other spondyloarthropathies associated with cutaneous disease is considered. PMID- 10467499 TI - An infectious mononucleosis-like syndrome induced by minocycline: a third pattern of adverse drug reaction. AB - A 22-year-old black man developed fever, chills, fatigue, night sweats, tender lymphadenopathy, and a generalized, pruritic, macular eruption 3 weeks after starting minocycline therapy for acne. His illness was also characterized by a palpable spleen tip, marked lower extremity and scrotal edema, and generalized lymphadenopathy. The patient had leukocytosis with a large percentage of atypical lymphocytes on peripheral smear and liver dysfunction. Titers for Epstein-Barr virus, hepatitis B, toxoplasmosis; and cytomegalovirus were all negative. Human immunodeficiency virus-1 viral load and antibodies were also negative. Marked improvement was noted after the discontinuation of minocycline and the use of systemic corticosteroids. With the negative viral serologies, the clinical picture was most consistent with an infectious mononucleosis-like syndrome produced by the minocycline ingestion. PMID- 10467500 TI - Acne mechanica due to an orthopedic crutch. AB - Acne mechanica describes local irritation due to the skin being pressured, occluded, rubbed, squeezed, stretched, or heated. Examples of various physical traumas that provoke lesions in acne patients are given. Users of orthopedic crutches are at particular risk for such conditions. A crutch is another extrinsic factor to be added to the list of causes of acne mechanica. PMID- 10467501 TI - White superficial onychomycosis of the fingernail caused by Trichophyton rubrum in an immunocompetent patient. AB - White superficial onychomycosis in immunocompetent patients is most commonly due to Trichophyton mentagrophytes, and almost exclusively involves the toenail. We present a unique case of an otherwise healthy patient with white superficial onychomycosis involving the fingernail caused by T. rubrum. PMID- 10467502 TI - Aplasia cutis congenita associated with fetus papyraceous. AB - This report describes a healthy infant born with an isolated, truncal cutis aplasia defect in association with a fetus papyraceous. Effective healing of the cutaneous defect occurred over the course of a month. A 2-year follow-up demonstrate stable wound coverage. This rare association of aplasia cutis congenita, with a fetus papyraceous points most likely to the vascular causation of the cutaneous defect. PMID- 10467503 TI - Treatment of pyoderma faciale with isotretinoin in a patient with ulcerative colitis. AB - The explosive onset of fluctuant facial papulonodules, usually in young women, is characteristic of pyoderma faciale. This disorder is neither a true pyoderma nor a variant of acne, but rather a severe form of rosacea. The most effective therapeutic modality appears to be isotretinoin, especially if preceded by a brief course of oral corticosteroids or a short interval of application of potent topical corticosteroids. Despite our concern about the potential adverse effects of systemic retinoids on underlying inflammatory bowel disease, isotretinoin was given to a patient with refractory pyoderma faciale. Response was dramatic, and no ill effects were encountered. PMID- 10467504 TI - Giant cutaneous horn associated with verruca vulgaris. AB - Cutaneous horn (cornu cutaneum) is the clinical description of a hyperproliferation of compact keratin in response to a wide array of underlying benign and malignant pathologic changes. We report a patient with a giant cutaneous horn associated with a verruca vulgaris. The possible causes of cutaneous horns are reviewed. PMID- 10467505 TI - Unresponsive eczematous dermatitis: a case of pancreatic cancer masquerading as cutaneous T cell lymphoma. AB - Skin diseases may be the presenting sign of malignancy, but strict criteria are required to make the diagnosis of a paraneoplastic syndrome. Common dermatoses may also herald an underlying malignancy without meeting these criteria. We report the case of an elderly man with an unresponsive eczematous dermatitis whose evaluation eventually revealed a pancreatic carcinoma. This case is presented to alert the clinician that a common dermatosis may be the first sign of an occult malignancy, especially if it behaves in an atypical or aggressive manner or is recalcitrant to standard therapies. PMID- 10467506 TI - Recalcitrant acne vulgaris secondary to a dental abscess. AB - This case report describes a middle-aged man with acne that was recalcitrant to numerous medications, including three courses of isotretinoin. His condition cleared after an infected tooth was removed and recurred when another tooth became carious. Acne vulgaris is associated with several immunologic responses including the production of antibodies against Propionibacterium acnes, the gram positive bacteria found in acne lesions. We believe that the presence of our patient's dental infections provoked a follicular inflammatory response resulting in his recalcitrant acne. PMID- 10467507 TI - Atypical presentation of herpes simplex virus in a patient with chronic lymphocytic leukemia. AB - Perianal infections caused by herpes simplex virus are common in immunocompromised patients. The cutaneous presentation in these patients is often atypical, overlaps with the clinical features of other diseases, poses a difficulty in diagnosis, and responds poorly to treatment. An immunocompromised patient with chronic lymphocytic leukemia, treated with oral corticosteroids, presented with chronic perianal ulcerations. This patient was referred for evaluation and treatment of "recalcitrant" pyoderma gangrenosum. Prompt diagnosis was possible when the clinical features were recognized and appropriate biopsy and cultures were obtained. We describe an atypical presentation of herpes simplex virus associated with both an endogenous and exogenous induced immunodeficiency, and stress the importance of routinely performing cultures on all perianal ulcerations and anal fissures to avoid the misdiagnosis, inappropriate treatment, and prolonged discomfort of these afflicted patients. PMID- 10467508 TI - Steatocystoma multiplex: a quick removal technique. AB - A 27-year-old man with multiple lesions located on his chest and neck and diagnosed as steatocystoma multiplex desired removal of these lesions. We report a facile, fast, and successful technique for the removal of lesions of steatocystoma multiplex. Using a no. 11 blade, we incised the domes of numerous lesions and removed the cyst walls with small artery forceps. Within 1 month, the incisions healed without scarring, and after 4 months of follow-up, they had not recurred. This procedure allows the removal of many lesions of steatocystoma in a few office visits. PMID- 10467509 TI - Extensive oral mucosal ulcerations caused by misuse of a commercial mouthwash. AB - This case report describes severe mucosal injuries following misuse of an undiluted over-the-counter mouthwash with a high alcohol content (70%), oil of peppermint and arnica. The mouthwash was to be diluted 5:1 with water. The patient used undiluted solution to better treat her self-diagnosed "contagious gum infection." She experienced burning sensation with each rinse and developed severe mucosal injuries subsequently. Her oral condition improved within 48 hours following discontinuation of use of the mouthwash and application of a mixture of Benadryl Elixir, Maalox Plain, and 2% viscous Lidocaine. A detailed history and review of a patient's medical condition will help to differentiate self-induced mucosal injuries from those caused by an allergic reaction or skin diseases. PMID- 10467510 TI - Generalized lichen nitidus: case report and literature review. AB - Lichen nitidus is a rare condition of unknown cause, characterized by minute, flesh-colored, shiny papules occurring on the genitalia, abdomen, chest, and extremities. This disorder is most often localized, but a few cases of generalized lichen nitidus have been reported. We describe a young patient with a 1.5-year history of unremitting generalized lichen nitidus. PMID- 10467511 TI - The influence of prosodic structure on the interpretation of temporary syntactic ambiguity by young and elderly listeners. AB - The contribution of prosody to the interpretation of temporary syntactic ambiguity was examined for young and elderly listeners using a sentence completion task. Temporary syntactic ambiguity refers to cases where it may be temporarily unclear whether a syntactic clause boundary has or has not been reached based on what has been heard in the sentence to that point. Results suggest that both young and elderly adults use a computationally less demanding late-closure parsing strategy whenever possible, but that sentence prosody can override this tendency when an alternative closure position is clearly signaled. Although subtle differences appeared in regard to sentence completion strategies and latencies to completion, results suggest that efficient resolution of syntactic boundary uncertainty and effective use of sentence prosody are two features of language comprehension that remain well-preserved in normal aging. PMID- 10467512 TI - Evaluating a measure of everyday problem solving for use in African Americans. AB - Results from previous research on everyday problem solving involving Caucasians suggests that it may be a useful concept in studying cognitive aging in African Americans. The purpose of this investigation was to examine: (1) the factor structure of an everyday problem solving in a sample of African Americans, (2) the internal consistency of everyday-problem solving in a sample of African Americans, and (3) the relationship of problem solving to demographic factors, physical functioning, and measures of fluid ability. The sample included subjects recruited from Baltimore, MD. The sample consisted of 249 community dwelling African-American adults with a mean age of 67.8 years (SD = 8.47). Variables included the Everyday Problem Solving Test (EPT), gender, age, education, physical functioning, and inductive reasoning. Everyday problem solving as a latent construct was confirmed and the split half reliability was high (.89). As in previous research, inductive reasoning and physical functioning were related to everyday problem solving abilities. We also found that certain domains of the EPT are more influenced by demographic factors than others. Our finding suggest that the Everyday Problems Test is appropriate for use with African American samples. PMID- 10467513 TI - An experimental and computational analysis of age differences in the recognition of fragmented pictures: inhibitory connections versus speed of processing. AB - Young and older adults were tested for the ability to identify degraded pictures presented either in a series of incremental steps with each step increasing the completeness of the visual information (ascending condition) or in one single exposure (fixed condition). Significant interference effects, indicated by a superiority of fixed over ascending presentations, appeared at a lower level of performance for the older adults than for the young adults. This finding was consistent with the notion of an inhibition deficit operating in normal aging. A computer simulation, based on simple connectionist architecture, demonstrated that an age-related inhibition deficit in the identification of fragmented pictures can be produced by slowed processing rates. PMID- 10467514 TI - Processing speed and adult age differences in activity memory. AB - Immediate and delayed recall of performed cognitive activities was examined in 136 adults aged 20 to 85. Hierarchical regression analyses were used to assess the association between perceptual speed and age differences in activity memory. The age-related variance in delayed activity recall was reduced by 52% by the statistical control of perceptual speed, and the age-related variance in immediate activity recall was reduced by 91%. Thus, adult age differences in delayed and immediate activity memory were found to be associated with limitations in perceptual speed. The cognitive effort that is required to perform cognitive activities may tax the processing resources of older adults, prohibiting successful encoding of the activities. PMID- 10467515 TI - Genetic and environmental influences on forced expiratory volume in midlife: a cross-cultural replication. AB - Previous research has shown that forced expiratory volume (FEV) is a useful predictor of remaining life in older adults. The present analyses are an attempt to replicate results from a study of Swedish twins which demonstrated substantial heritability of FEV with a sample of middle-aged Russian adult twins. Data were collected from 116 pairs of Russian twins (monozygote = 71, dizygotic = 45, mean age = 40.9 years). Phenotypic correlations between FEV, age, gender, height, and cigarette consumption (in pack years) were all significant, ranging from -.72 to .31. After the effects of age, gender, height, and smoking were partialled out of FEV, quantitative genetic analyses were conducted. Shared environmental effects were significant, accounting for 47% of the variance in FEV. Genetic effects, which accounted for about 28% of the variance, could be dropped from the model without a significant decrease in the fit. These results are discussed in relation to previous research conducted in other countries. PMID- 10467516 TI - Effects of age on short-term memory for time in rats. AB - Working memory (WM) for event duration was investigated in four different age groups of rats (6, 12, 18, and 24 months), using a delayed, symbolic-matching-to sample procedure. In an initial experiment, all age groups responded as though a long sample duration were a short one when retention intervals (RI) were applied. This effect is called the choose-short effect. In a second experiment, in order to test whether, in compliance with the subjective-shortening model, there would be a "shortening" of the sample duration according to the length of the RI, the psychophysical function relating the probability of choosing the long comparison stimulus to sample duration and the point of subjective equality (PSE) were determined across successive RIs in two age groups (6 and 18 months). A significant shift of the psychophysical function towards a longer duration and a significant increase of the PSE appeared as a function of the RI in 6-month-old, but not in 18-month-old rats. These results support the idea that the choose short effect is due to a shortening of the event duration in the WM in young animals, whereas there is no evidence that the sample forgetting occurred on a time dimension in older rats. PMID- 10467517 TI - Characterization of ATP-sensitive potassium channels in human corporal smooth muscle cells. AB - Potassium (K) channels play a significant role in modulating human corporal smooth muscle tone, and thus, erectile capacity. Recent pharmacological studies indicate that the metabolically-regulated K channel (KATP) may be an important modulator of human penile erection with significant therapeutic potential. The goal of these initial studies, therefore, was to utilize patch clamp techniques to characterize the putative KATP subtype(s) present in cultured and freshly isolated human corporal smooth muscle cells. In the cell-attached patch mode, two distinct unitary K+ currents were identified whose respective conductance values were similar in cultured and freshly isolated smooth muscle cells. In cultured myocytes, the measured conductance values in symmetric KCl (140 mM) solutions were 59.1 +/- 2.7 pS and 18.4 +/- 2.1 pS (n = 5 cells). Under identical experimental conditions in freshly isolated myocytes, corresponding conductance values were 59.2 +/- 3.7 pS and 18.5 +/- 2.4 pS, respectively (n = 4 cells). I-V curves constructed during step depolarization (-60 to +80 mV), revealed a linear I-V relationship for both unitary conductances. Single channel records documented that both conductances were reversibly inhibited by the application of ATP (1-3 mM) to the bath solution in the inside-out attached patch configuration. The unitary activity of both K channel subtypes was significantly increased by the application of pinacidil (10 microM) and levcromakalim (10 microM). Whole cell patch recordings documented a glibenclamide-sensitive, pinacidil- and levcromakalim-induced increase in the whole cell outward K+ current during step depolarization (-70 mV to +130 mV) of 105 +/- 37%, 139 +/- 42%, respectively. These data confirm and extend our previous observations, and provide the first evidence for the presence of KATP channel subtypes in human corporal smooth muscle cells. PMID- 10467518 TI - Prostaglandin E1 activates the large-conductance KCa channel in human corporal smooth muscle cells. AB - The large conductance calcium-sensitive potassium channel (KCa or maxi-K) is an important modulator of human corporal smooth muscle tone, and therefore, erectile capacity. The goal of this investigation was to evaluate the actions of prostaglandin E1 (PGE1), the most widely used and effective drug for the treatment of impotence, on the activity of the KCa channel, a prominent K+ current present in human corporal smooth muscle. Whole-cell patch clamp studies conducted on short-term cultured and enzymatically dissociated human corporal smooth muscle cells, revealed mean resting potentials of -50.8 +/- 2.1 mV (n = 8) and -34 +/- 4 mV (n = 8), respectively. In the attached-patch configuration, the corresponding single-channel slope conductance values for the KCa channel subtype were 173 +/- 4 pS (n = 8) in cultured cells, and 190 +/- 13 pS (n = 3) in freshly isolated myocytes. Furthermore, voltage clamp experiments revealed that relative to control values, the application of PGE1 to cultured cells (3.3 or 33 microM) elicited an apparent increase in both the open probability (Po; ranging from 1.2 23 fold), and the mean open time (5-6 fold) of the KCa channel at membrane potentials of +90 mV and +110 mV. PGE1-induced alterations in KCa channel activity were also observed in freshly isolated corporal myocytes. In the whole cell-recording mode, statistically significant, Charybdotoxin-sensitive (100 nM) 2-3 fold increases in the outward K+ currents were observed in both cultured and freshly isolated corporal myocytes. The presence of a PKA inhibitor (fragment 6 22 amide; 10 microM) in the pipette tip was also associated with a nearly complete ablation of the observed PGE1-induced whole cell K+ currents. Taken together, these data confirm and extend our previous observations, and indicate that PGE1-induced relaxation of human corporal smooth muscle is related, at least in part, to activation of the KCa channel subtype resulting in cellular hyperpolarization. PMID- 10467519 TI - Differential gene expression of growth factors in young and old rat penile tissues is associated with erectile dysfunction. AB - The molecular mechanisms of erectile dysfunction with aging are unclear. Recent studies have suggested that growth factors may play a role in the etiology of erectile dysfunction. This present study was designed to test the hypothesis that gene expression of various growth factors such as TGF alpha, TGF beta 1, TGF beta 2, TGF beta 3, IGF and NGF modulate with aging in rat penile tissues. For this purpose, total RNA was extracted from young and old rat penile tissues and the gene expression for these growth factors was determined by differential reverse transcriptase-polymerase chain reaction (RT-PCR) using specific oligonucleotide primers. mRNA levels of growth factors were quantified by using beta-actin as an internal standard. The results of these experiments suggest that: (1) young and old rat penile tissues expressed mRNA transcripts for TGF alpha, TGF beta 1, TGF beta 2, TGF beta 3, IGF and NGF; (2) TGF beta 1 gene expression was significantly increased in old rat penile tissues as compared to young; (3) mRNA transcripts for NGF and TGF beta 3 were significantly lower in old rat penile tissues as compared to young; and (4) TGF alpha, TGF beta 2 and IGF mRNA expression did not change in young and old rat penile tissues. These results suggest that the differential gene expression for various growth factors in young and old rat penile tissues may be important in understanding the pathophysiology of erectile dysfunction associated with aging. PMID- 10467520 TI - Stimulation of collagen production in an in vitro model for Peyronie's disease. AB - AIMS OF THIS STUDY: This study evaluated whether human cavernosal myofibroblasts in cell culture is a viable model for the study of the role of oxygen free radicals in the production of collagen types I and III, as observed in Peyronie's disease. METHOD: Human cavernosal cells in primary culture were incubated with 3H proline in the absence or presence of (i) glyceraldehyde; (ii) alpha-tocopherol (vitamin E); (iii) a combination of the two; or (iv) gamma interferon alone or in combination with glyceraldehyde. Collagen production was monitored after precipitation by specific monoclonal antibody and quantitated using a scintillation counter. RESULTS: Collagen type III was stimulated to higher than baseline values after doses of 10 and 100 microM glyceraldehyde was added and showed suppression of stimulation with incorporation of alpha-tocopherol. There was a 40% increase in collagen type III production as compared to baseline values in glyceraldehyde-treated cells. Collagen type I showed no consistent stimulation or suppression. In glyceraldehyde-stimulated transformed caveronsal cells, alpha tocopherol treatment caused a 10-60% decrease in collagen type I and III production. With the addition of 100,000 IU/ml gamma interferon, a significant reduction of both collagen types I and III was observed. CONCLUSIONS: The generation of oxygen radicals is associated with the stimulation of collagen production in cavernosal cells. Transformed fibroblasts from cavernosal cells in culture can be utilized to explore possible etiologies of Peyronie's disease and to further evaluate potential medical therapies for this pathological condition. PMID- 10467521 TI - Interdisciplinary assessment and follow-up of patients with erectile dysfunction- psychiatric aspects. AB - The objectives of this investigation were to assess the psychiatric comorbidity and patients' satisfaction with treatment in a sample of 73 men with erectile dysfunction (ED) who were consecutively admitted to an andrologic outpatient centre. After comprehensive interdisciplinary assessment, 49% of these patients were assigned an organic etiology, 33% had psychogenic impotence, and in 18% somatic and psychogenic factors seemed to be relevant. 63% of all patients received a psychiatric diagnosis (according to ICD-9). Depressive disorders were found in 25%. Organic psychosyndromes (nonpsychotic) were prominent in the somatic group. In a follow-up after 29.4 +/- 5.0 months, 41 out of 73 patients could be reevaluated, of which 85% of organic patients had received some kind of therapy, but 65% were not satisfied with the chosen treatment. These results underline the importance of close interdisciplinary co-operation in the assessment and therapy of patients with erectile importance. Potential reasons for drop-out and limited acceptance of the offered treatments are discussed. PMID- 10467522 TI - 5-HT4 receptors in isolated human corpus cavernosum? AB - The novel serotonin subtype-4 (5-HT4) receptor agonist, SC53116 (SC), produced a limited relaxation of noradrenaline (NA) pre-contracted human corpus cavernosum (CC) smooth muscle in vitro. This effect was not significantly attenuated by the 5-HT4 antagonist SDZ250557 (SDZ). In the presence of (+/-) pindolol (1 microM) and methysergide (1 microM), employed to mask 5-HT1 and beta-adrenergic, and 5 HT2 receptors respectively, SC failed to relax NA pre-contracted CC strips to a greater extent than saline. Functional cAMP dependent relaxation pathways were demonstrated by a significant reduction in NA induced tone by prostaglandin E1 (PGE1) and isopropylnoradrenaline (IPNA), the action of the latter compound was effectively eliminated in the presence of (+/-) pindolol. Relaxation of NA induced tone caused by the nitric oxide donor nitro-glycerine (NTG) was significant and similar in the absence and presence of the 5-HT and beta adrenergic antagonists. The results of this present study indicate that human corporal smooth muscle does not contain 5-HT4 receptors and that, although compounds like SC act to relax non-vascular smooth muscle via cAMP dependent mechanisms, 5-HT4 receptor agonists may be expected to be of limited utility in triggering cAMP dependent relaxation responses in human CC. PMID- 10467523 TI - The role of growth factor on regeneration of nitric oxide synthase (NOS)- containing nerves after cavernous neurotomy in the rats. AB - Nitric oxide synthase (NOS) containing nerve regeneration can be seen six months after unilateral cavernous nerve neurotomy in rats. However, its molecular mechanism is still unknown. It is believed that growth factors are involved in this phenomenon. In this study we investigated the change of NOS containing nerve fibers and the RNA expression of insulin like growth factor (IGF)-I, nerve growth factor (NGF), transforming growth factor (TGF)-alpha, TGF-beta 1, TGF-beta 2. TGF beta 3 and NOS on the penis after cavernous nerve neurotomy in rats. Male rats were divided into three groups: (1) sham operation (N = 10); (2) unilateral neurotomy of a 5 mm segment of the cavernous nerve (N = 15); and (3) bilateral neurotomy (n = 15). Electrostimulation of the intact cavernous nerve or pelvic ganglion was performed at one, three and six months. Nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase staining was used to identify NOS in the penile nerve fibers. The gene expression for growth factors and bNOS was investigated in corporal tissue by reverse transcriptase-polymerase chain reaction (RT-PCR) using specific oligonucleotide primers. One month after neurotomy, both unilateral and bilateral neurotomy groups showed a significant decrease in NOS-containing nerve fibers on the dorsal and intracavernosal nerves on the side of neurotomy, and a significantly lower mRNA expression of bNOS, IGF I and TGF-beta 2. At three months, the number of NOS-containing nerve fibers in the unilateral neurotomy group increased only slightly but at six months those in the intracavernosal nerve increased in a significant amount (P < 0.0001), however mRNA expression of bNOS, IGF-I and TGF-beta 2 showed a significant increase as early as at three months. After bilateral neurotomy, the NOS-positive nerve fibers in the dorsal and intracavernosal nerve were significantly decreased at one month and remained so at six months; no erectile response could be elicited by pelvic ganglion stimulation. In the unilateral neurotomy group at six months, more NOS-positive neurons in the pelvic ganglia were found on the intact side than on the side of the neurotomy (P < 0.003), indicating that the regeneration derives from pelvic ganglion neurons on the intact side. Furthermore, electrostimulation in the unilateral neurotomy group revealed a greater maximal intracavernosal pressure and a shorter latency period at six months than at one month (P < 0.014, P < 0.001, respectively). These data suggest that IGF-I and TGF beta 2 may play a key role in regeneration of NOS-containing nerve fibers in the dorsal and intracavernosal nerves after unilateral cavernous nerve injury. PMID- 10467524 TI - Spontaneous breakage of malleable prosthesis. PMID- 10467525 TI - Male erectile dysfunction. PMID- 10467526 TI - [Infection and vascular purpura]. AB - Palpable purpura is the hallmark of cutaneous vasculitis. Small-vessel vasculitis is a common vasculitis manifestation associated with acute or chronic infection. It is also characteristic of a systemic disease whether infectious or not. The pathogenic mechanisms appear to be complex: immune complex formation, vessel damage or altered vessel function mediated directly by infectious agents, humoral or cellular immunologic response. It is also a reaction to mixed cryoglobulinemia. Diagnosis of cutaneous vasculitis is simple (palpable purpuric eruption, nodules, vesiculobullous lesions, ulcerations), but etiological investigation is often difficult because the infectious origin is only rarely demonstrated. This type of purpura occurs in bacterial endocarditis and therefore blood cultures must be performed in any febrile patient particularly in the presence of a cardiac murmur. In fact the viral, parasitic or bacterial infectious origin is demonstrated in less than 30% of the cases of leucocytoclastic vasculitis. While focal sepsis is often found and its eradication should be followed-up, its role has not been proven particularly as antibiotics alone themselves can cause hypersensitivity vasculitis. Finally, mention must be made of virus induced vasculitis (B and C hepatitis, cytomegalovirus, parvovirus), antiviral treatment which permits better control of vasculitis. PMID- 10467527 TI - [Treatment of venous thromboembolic disease in the elderly]. AB - The incidence of deep vein thrombosis and pulmonary embolism is higher in the elderly due to the greater frequency of risk factors among this age group. Classic treatment with heparin and subsequently oral anticoagulant is still the most commonly used. Older patients may be at increased risk for anticoagulant related bleeding for several reasons: increased anticoagulant effect of warfarin, increased prevalence of comorbidity and incidence of adverse drug reactions. As well as the usual contra-indications to such treatment, the psychological and physical well being of the patient must be assumed before an oral anticoagulant can be given. Careful prescribing is required: a low starting dose, a strict monitoring regime, for a limited duration. The indications for use of an inferior vena cava filter are wider in the older age group, not only for those in whom heparin is contraindicated, or has failed, but also for those who require treatment indefinitely with contra-indications to oral anticoagulant. Careful consideration of risk factors and the use of an individually designed prophylactic treatment are the best way to tackle this difficult problem in the elderly person. PMID- 10467528 TI - [Modulation of angiogenesis. A new therapeutic tool for vascular diseases]. AB - Angiogenesis permits the growth of new vessels. This process may be modified during the course of some pathological phenomenon, i.e. cancer, atherosclerosis or diabetes. After a brief description of angiogenesis pathophysiology, the therapeutic possibilities in cardiovascular diseases are discussed. Initial results of therapeutic trials in coronary and peripheral artery diseases are detailed, and perspectives for the future are reviewed. PMID- 10467529 TI - [Infections of vascular prostheses of the abdominal aorta. Diagnostic and therapeutic problems]. AB - Aortic graft infection is one of the most serious complication of vascular surgery. While the incidence of such infections has declined over the years, it continues to be associated with high mortality and high rates of limb loss. Graft implanted in the inguinal area have a higher rate of infection than those that lie entirely within the abdomen. Infection of the intraabdominal extremities of vascular grafts is most frequently associated with prostheto-enteric fistula. DIAGNOSIS AND TREATMENT: Despite the many available sophisticated imaging techniques diagnosis remains difficult, particularly with intra-abdominal grafts because of nonspecific clinical and imaging findings. A variety of approaches to aortic graft infection have been proposed and optimal treatment usually requires an association of systemic antibiotic administration with surgery. Removal of the entire infected graft is often necessary for cure and there are only few situations in which conservative treatment is acceptable. A careful identification of the infecting microorganisms and in vitro susceptibility testing are essential for successful therapy. PREVENTION: Prevention consists in a strict adherence to principles of asepsis and the use of prophylactic antibiotics in vascular surgery. The use of antibiotics-bound knitted grafts has been recently proposed. PMID- 10467530 TI - [Hemorrhagic complications of antivitamin K. Report of 75 hospitalized patients]. AB - Hemorrhagic complications are the most frequent complications of antivitamin K (AVK) treatments and can be life-threatening. We report 75 patients from a University Hospital. They were 40 males and 35 females (median age 74 years, 20 94), and were classified into 3 grades according to clinical picture: grade 1 (no surgery or transfusion, grade 2: surgery or blood transfusion needed, grade 3: death). 43 patients had grade 1 complications, 27 grade 2, and 5 grade 3 complications. The most frequent complications were muscular hematomas (36 patients), sub-cutaneous hematomas (14 patients), digestive bleeding (13 patients), hematuria (12 subjects). Eight patients had intracerebral bleeding, of whom 3 died. The treatment time was very variable (1 to 988 weeks). Only half patients had a prothrombin rate (PR) below 20% but two thirds had an INR above 5. This study showed that PR was a poor predictor of hemorrhagic complications. INR was a better parameter. For 15 patients, we considered that the indication was unadapted or questionable, among whom 2 died. This work suggests that the promotion of AVK prescription rules should go on. PMID- 10467531 TI - [Consequences of obliterating arteriopathy of the legs on the professional activity of patients and external assistance]. AB - Peripheral arterial occlusive disease of the lower limbs results in increased risks of cardiovascular and cerebrovascular morbidity and mortality. However, peripheral arterial disease also has an incapacitating effect upon patients' daily lives, and indeed can have considerable consequences upon their quality of life as well as upon their professional activities. The purpose of this study was to describe patients in terms of their professional activities, the consequences of the disease upon such activities, need for external assistance, and the effects of treatment over a six-month treatment follow-up period. This was a prospective, randomised, double-blind, multicentre study comparing naftidrofuryl (three 200-mg tablets per day) versus placebo in patients presenting with stage II peripheral arterial disease. Four study visits were scheduled for each patient: an initial selection visit, followed by a 30-day placebo wash-out period, an inclusion visit (to allow verification of inclusion and non-inclusion criteria), and follow-up visits after 3 and 6 months of treatment. 234 patients were included in the study in intention to treat (117 patients in the naftidrofuryl group and 117 patients in the placebo group). These two groups were entirely comparable for all criteria (sociodemographic criteria, risk factors, cardiovascular history, duration of disease) except for obesity and dyslipidaemia, for which prevalence was higher in the naftidrofuryl group. 12.4% of the study population were professionally active at the time of the survey (mean age: 65.9 years). However, 42.3% of patients had in fact been active at the time when arterial disease became manifest. Of these patients, 45.4% (45 patients) stated that arterial disease had affected their professional activity: 28.9% (13 patients) reported a change in their activity, 26.7% (12 patients) reported partial suspension of their activity, and 44.4% (20 patients) stated that all professional activities had ceased. Changes in professional activity comprised invalidity (36.2%), prolonged sick leave (25.5%), premature retirement (14.9%), sick leave (17%), unemployment (6.4%), reduction in long-distance travel (4.3%), change of job (8.5%) or salary reduction (2.1%). During the 6-month follow-up period, only 4 patients were on sick leave (3 in the placebo group and 1 in the naftidrofuryl group). Furthermore, less than 10% of the study population required external assistance as a result of the disease. This analysis thus highlights the repercussions of this disease upon the patients' daily and professional activities. PMID- 10467532 TI - [Effectiveness and safety of hydroxyethyl-rutosides in the local treatment of symptoms of venous insufficiency during air travel]. AB - During air travel, the length of time spent in a sitting position and the absence of muscular activity in the calves severely slow the rate of blood flow in the lower limbs. The aim of this randomized, cross-over, double-blind study was to evaluate local application of Hydroxyethyl-rutosides (O-Beta Hydroxyethylrutosides) in the treatment of symptoms of venous insufficiency including stasis-induced edema during extended air travel on flights exceeding 6 hours. Hydroxyethyl-rutosides or placebo was applied every 3 or 4 hours throughout the flight. In the 51 subjects evaluated (both males and females) the results show statistically significant differences favoring treatment with Hydroxyethyl-rutosides both with regard to objective signs of edema: change in minimum ankle circumference was less during trips in which Hydroxyethyl-rutosides was applied, whether compared with the maximum measurement (p = 0.04) or the last measurement made during the flights, and with regard to subjective signs: several symptoms occurred significantly less frequently when the subject applied Hydroxyethyl-rutosides during the flight [pain (p = 0.03), sensation of heavy and tired legs (p = 0.04) and sensation of swelling (p = 0.02)]. the patient's overall assessment of the treatment was also favorable after using Hydroxyethyl rutosides Gel (p = 0.01). the number of subjects complaining of edema (pitting edema, marks of shoes, difficulties putting shoes back on) was significantly lower during periods of treatment with Hydroxyethyl-rutosides Gel (p = 0.001). Local application of Hydroxyethyl-rutosides, 3 to 4 times during 6 to 14 hours is thus effective in treating the main symptoms of venous insufficiency including stasis-induced edema caused by extended periods in the sitting position during long air flights. PMID- 10467533 TI - [Behcet's disease and iatrogenic arterial complications: importance of pre operative noninvasive explorations]. AB - We report a case of Behcet's disease complicated with a popliteal aneurysm, which appeared 8 years after first etiologic diagnosis. The points of interest of this observation are the pre-operative explorations and particularly the Magnetic Resonance Angiography (RMA). This last seems to be particularly safe for aortic and lower limbs aneurysms explorations, and can be performed without arterial functions. In fact the risk of arteriogram is effective in Behcet's disease with secondary false aneurysms. RMA and US-Doppler scan must be first proposed in Behcet's disease in case of aortic and peripheral aneurysms. PMID- 10467534 TI - [Behcet's disease with multiple intracranial arterial aneurysms. Report of a case]. AB - Behcet disease is a rare condition in central Europe but more common in Morocco. A case of multiple intracranial arterial aneurysms occurring in a 44 year-old Moroccan patient with 2-years history of Behcet's disease is reported. CT-scan showed an infarction in the right middle cerebral artery territory. Panangiography showed sacciform aneurysms of the bifurcation of the right and left middle cerebral arteries. The draining veins and sinuses were normal. The two aneurysms were successfully clipped by two microsurgical frontotemporal approach in one surgical time. There have been only eight reports of intracranial arterial aneurysms associated to Behcet disease in the literature. PMID- 10467535 TI - [Paraplegia secondary to surgery of the abdominal aorta]. AB - We report the case of an 84 year-old man who suffered of paraplegia after surgery of an abdominal aortic aneurysm. The neurologic status did not improve during 5 years of follow-up. This complication is rare in light of the review of the literature. It is more frequent after emergency (1.4%) and redux (19%) surgery than after elective surgery (1.1%). Its mechanisms are anatomic (occlusion of the Adamkievitz artery and of the pelvic collateral arteries) and hemodynamic mechanisms (thromboembolism, low systemic tension, long supra-renal clamping). Since pre-operative aortography to detect anatomic variations is not realistic, its only prevention remains maintenance of intra-operative hemodynamic, systemic heparinization, and preservation of the pelvic collateral circulation. PMID- 10467536 TI - [Echo-doppler surveillance of revascularization of the legs in asymptomatic postoperative patients. Why? With what rhythm? How much time? How?]. AB - AIM: Routine duplex surveillance of patients following an arterial operation includes the scanning of symptomless patients. While the benefits of preventive correction of deteriorations, stenoses or dilatations, following aortic or lower limb arterial reconstruction are undisputed, there is no hard data on the effectiveness of this surveillance in terms of limb salvage, on its cost, and on the optimal frequency and duration. METHODS: The AA review the literature on this subject and suggest a programme which reconciles the patients' interests with the best use of health care financial resources. RESULTS: The possible complications depend on the type of operation. Some are symptomless. They occur mainly during the first year and less frequently, but regularly so, during the subsequent years. There is therefore no time limit for the duration of the surveillance. The effectiveness of this surveillance is low and thus the examination must be meticulous and its frequency adapted to the risks. Broadly speaking: During the first year, aorto-ilio-femoral reconstructions must be screened at one month, six months and one year; by-passes below the groin with no particular risk at six months and one year; by-passes at risk and those anastomosed to a lower leg or ankle artery require a further check-up at one month and three months. After the first year, aorto-ilio-femoral and similar by-passes must be checked annually; by passes below the groin, when at risk, must be checked annually; in those without any particular risk, a check-up every two years seems to be sufficient. Any patient found to have a deterioration on screening must be checked every six months. CONCLUSION: Careful screening is the only way of discovering a symptomless deterioration before the development of serious complications. As every arteriole reconstruction should be followed by a lifelong surveillance, in times of financial constraints the frequency of the check-ups must be tailored according to the risks present and the quality of the surveillance must be faultless. PMID- 10467537 TI - [Professional recommendations for follow-up of the type 2 diabetic patient excluding the follow-up of complications. National Agency for the Accreditation and Evaluation of Health]. PMID- 10467538 TI - Infection control team in the operating room: separating aspiration from reality! PMID- 10467539 TI - Worthwhile infection control information? A report from the 38th Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC), San Diego, California: 24-27 September 1998. AB - A review is presented of some of the presentations delivered at ICAAC 1998. These cover the subjects of handwashing, occupational infections risk exposure for healthcare workers and alterations in catheter practice to reduce both intra venous and urinary infections. PMID- 10467540 TI - Intrinsically vancomycin-resistant gram-positive organisms: clinical relevance and implications for infection control. AB - Intrinsic resistance to vancomycin in gram-positive bacteria presumably predates acquired vancomycin resistance in enterococci but it has only recently generated interest. Intrinsically resistant enterococci possessing the vanC gene and the non-enterococcal genera Leuconostoc, Lactobacillus, Pediococcus and Erysipelothrix are known to cause human infection. This review examines the available data on their identification, resistance mechanisms, epidemiology, clinical infections and antimicrobial susceptibility. Intrinsically vancomycin resistant gram-positives are usually opportunistic pathogens. Although serious infections may occur, treatment options remain available. No additional infection control measures for the intrinsically resistant genera appear justified with currently available evidence, although vigilance should be maintained to detect future changes in susceptibility patterns. PMID- 10467541 TI - Glove powder: implications for infection control. AB - Gloves are increasingly promoted for use by healthcare workers, but this use is not without risk. Data associating powdered gloves with an increased risk of latex allergy is available and there is circumstantial evidence that the powder used may increase bacterial environmental contamination. In animal models, corn starch, the material used as glove powder, promotes wound infection. Infection control teams need to be aware of this evidence and should support switching from use of powdered to powder free gloves. PMID- 10467542 TI - Inconsistent correlation between aerobic bacterial surface and air counts in operating rooms with ultra clean laminar air flows: proposal of a new bacteriological standard for surface contamination. AB - The relationship between surface contamination (cfus/m2/h) with particles carrying aerobic bacteria and corresponding air contamination rates (cfus/m3) was evaluated in operating rooms (OR) equipped with ultra clean vertical or horizontal laminar airflow (LAF). For the evaluation we collected data during strictly standardized sham operations using non-woven disposable or cotton clothing. Air contamination in the wound and instrument areas (Casella slit sampler) was related to the surface contamination rate (settle plates) in the same areas and in addition, on the patient chest. Typically, the mean surface counts were 20-70 cfus/m2/h and the air counts 1-2 cfus/m3 in disposable clothing experiments, whilst the use of cotton clothing resulted in higher counts of 100 200 cfus/m2/h (wound P > 0.05, patient P > 0.05, instruments P < 0.01) and 4 cfus/m3 (P < 0.02-0.001). In the vertical LAF, taking both disposable and cotton clothing operations together, the surface and air contamination rates (surface/air ratio SAR) were highly correlated (P = 0.02-0.004) and the ratio varied between 18:1 and 50:1 with a mean for wound air of 36:1. Using only disposable clothing in the vertical LAF, the number of significant correlations was reduced. With cotton clothing experiments in vertical LAF and in the horizontal LAF using disposable clothing, no significant correlation between surface and air contamination was found. The wide variation of SAR values and the inconsistent relationship between surface and air counts indicates that measurement of OR air contamination represents an unhelpful method for assessment of surgical site contamination in LAF units. We propose instead that colony counts on sedimentation plates is a clinically more relevant indicator of bacterial OR contamination in LAF units. In addition to the current bacteriological standard for ultra clean OR air of (< 10 cfus/m3) we suggest a corresponding standard for the surface contamination rate of < 350 cfus/m2/h. PMID- 10467543 TI - Nosocomial outbreak of gentamicin-resistant Klebsiella pneumoniae in a neonatal intensive care unit controlled by a change in antibiotic policy. AB - Between August and November 1997, a nosocomial outbreak caused by gentamicin resistant Klebsiella pneumoniae occurred in the Neonatal Intensive Care Unit (NICU) of our hospital. Thirteen neonates became colonized and three of them became infected. Comparison of the isolates by amplified fragment length polymorphism (AFLP) revealed clonal similarity for isolates of eight neonates (homology > 90%). Cultures from environmental specimens were negative for gentamicin-resistant K. pneumoniae. A case-control study was conducted to identify risk factors associated with acquisition of gentamicin-resistant K. pneumoniae. Risk factors were low gestational age and birth weight. These neonates need more care and handling and may therefore, be more at risk of colonization. Length of stay on the NICU was significantly longer for cases, but mean time until colonization (6.3 days) was shorter than the total stay for controls (9.5 days). No single member of the medical or nursing staff was significantly more involved with cases than with controls. The outbreak was stopped by replacing gentamicin by amikacin as the antibiotic of first choice whenever the use of an aminoglycoside antibiotic was indicated. PMID- 10467544 TI - Cost of antimicrobial treatment for nosocomial infections based on a French prevalence survey. AB - An estimate of the antibiotic cost of nosocomial infections (NI) was made in a university hospital group based on data collected in adult inpatients enrolled in the French national prevalence survey in 1996. Among the 6839 study patients, 636 (9.3%) presented with at least one NI, of these, data on antimicrobial treatment were available for 480. The overall daily antibiotic cost was estimated between FF 49,439 and 103,526, resulting in FF 103 to 216 per infected patient. The most expensive antibiotic treatment was prescribed in intensive care patients, for pneumonia for device-related NI, or for multi-resistant bacterial infections. Non documented NI represented about 20% of the overall antibiotic cost. Beta-lactam antibiotics, especially third generation cephalosporins, and parenteral fluoroquinolones were the most expensive antimicrobial drugs. The cost of antibiotic treatment for NI represents a significant part of hospital expenditure that should be reduced by better control of highly expensive prescriptions. PMID- 10467545 TI - The prognostic factors of cryptococcal meningitis in HIV-negative patients. AB - Seventy-one patients with cryptococcal meningitis, 46 males and 25 females, aged 15-83 years, were included in this study. Their initial clinical manifestations, cerebrospinal fluid (CSF) features, and therapeutic results were analysed. Patients were treated with three different regimens: amphotericin B, fluconazole, and combination therapy. Based on the therapeutic results, the 71 patients were also divided into cured, improved, and failed groups. For statistical comparison, the clinical manifestations and CSF features, were compared according to therapeutic outcome. There was no statistical difference in outcome among the three different antifungal regimens. However, patients treated with fluconazole required 36% fewer days of hospitalization compared with those receiving amphotericin B. Significant prognostic factors, included low CSF glucose, high CSF lactate, high CSF cryptococcal antigen titre (> or = 1:1024), initial level of consciousness, the presence of seizure, hydrocephalus, and central nervous system vasculitis. Multiple logistic regression analysis showed that only initial level of consciousness, and CSF antigen titre were strongly associated with therapeutic failure after other potentially confounding factors were adjusted for. Because some of the prognostic factors in cryptococcal meningitis can be corrected, early diagnosis, early use of appropriate antifungal treatment, and the correction of the underlying metabolic derangements are important in management. PMID- 10467546 TI - Usefulness of genotyping with microsatellite markers to investigate hospital acquired invasive aspergillosis. AB - To assess whether invasive aspergillosis (IA) was hospital acquired, Aspergillus fumigatus isolates, obtained during a one-year study from inpatients with haematological diseases and IA and from their environment, were genotyped using microsatellite markers. The analysis of 62 environmental isolates showed an extremely diverse A. fumigatus population with 43 genotypes represented only once. Eight genotypes were found more than twice at different times and/or at different locations showing that a given isolate can persist over time and is not dependent on a specific location. Twenty-seven isolates were obtained from 12 patients with IA. Of eight patients with multiple isolates, four were infected with isolates of different genotypes. Five patients (42%) had hospital-acquired IA according to the following definitions: patients infected with an isolate found in the environment, or patients infected with the same genotype. Although genotyping results are highly suggestive of hospital-acquired IA, this cannot be proved definitively because of the high diversity of the A. fumigatus population and the limited environmental sampling. A better knowledge of the A. fumigatus population outside hospitals is needed. For this purpose, genotyping using microsatellite markers seems appropriate. PMID- 10467547 TI - A new technique for quantitative bacterial assessment on burn wounds by modified dermabrasion. AB - Bacterial colonization and invasive bacterial infection is still one of the major problems in the treatment of burn victims. The standard procedures of bacterial monitoring of the burn would are i) swab-culture which is non-invasive but detects bacteria at the very surface and ii) biopsy-culture which gives a more complete view but has the disadvantage of being invasive. Therefore we developed a new technique for examination of microbial colonization of the wound surface. Dermabrasion of the upper layers of the wound was performed using a small rotating carbon-steel disc of defined roughness. The tissue samples obtained were analysed for bacterial growth in different culture media. Results were qualitatively and quantitatively compared with those of standard techniques performed in parallel. Our results show that this new technique is superior to the swab culture in identifying different bacterial species. The results can be compared with the biopsy technique, but has the advantage of being less invasive. PMID- 10467548 TI - Unnecessary hospital infection control practices in developing countries. PMID- 10467549 TI - The optimum frequency of ventilator circuit changes in mechanically ventilated children treated in the paediatric intensive care unit (PICU) PMID- 10467551 TI - Clinical governance and orthopaedic infection rate. PMID- 10467550 TI - Contaminated lithium heparin bottles as a source of pseudobacteraemia. PMID- 10467552 TI - Comparing perceived self-efficacy among adolescent Bosnian and Croatian refugees with and without posttraumatic stress disorder. AB - To examine the relationship between posttraumatic stress disorder (PTSD) and perceived self-efficacy (PSE), 98 Bosnian and Croatian refugees, ages 13-18, completed Bandura's Children's Multidimensional Scales of Perceived Self Efficacy, which measure self-predicted social functioning in 9 areas. Through interviews, participants were categorized according to DSM-IV criteria as traumatized PTSD-positive, traumatized PTSD-negative, and nontraumatized. ANCOVAs showed PTSD-positive participants exhibit higher PSE than nontraumatized participants in 5 of 9 areas. No significant differences among 3 female groups were observed; however, nontraumatized boys demonstrated lower PSE than the 2 traumatized groups in 7 of 9 areas. Thus, PTSD did not have a negative impact on PSE in this cultural context. Furthermore, surviving traumatic experience and preserving social support networks may be protective factors for maintaining high levels of PSE. PMID- 10467553 TI - Pathways from war trauma to posttraumatic stress symptoms among Tamil asylum seekers, refugees, and immigrants. AB - Path analysis was used to examine the antecedents of posttraumatic stress (PTS) symptoms in Tamil asylum-seekers, refugees, and immigrants in Australia. The Harvard Trauma Questionnaire and a postmigration living difficulties questionnaire were completed by 62 asylum-seekers, 30 refugees, and 104 immigrants who responded to a mail-out. Demographic characteristics, residency status, and measures of trauma and postmigration stress were fitted to a structural model in PTS symptoms. Premigration trauma exposure accounted for 20% of the variance of PTS symptoms. Postmigration stress contributed 14% of the variance. Although limited by sampling constraints and retrospective measurement, the study supports the notion that both traumatic and posttraumatic events contribute to the expression of PTS symptoms. PMID- 10467554 TI - Posttraumatic stress disorder and traumatic brain injury: are they mutually exclusive? AB - It has been suggested that posttraumatic stress disorder (PTSD) and traumatic brain injury (TBI) must be mutually incompatible disorders. However, growing empirical evidence has begun to question this. Evidence suggests that although PTSD may be relatively rare among the TBI population, some TBI patients seem to develop PTSD. We suggest two theoretical routes through which PTSD might develop in TBI patients: through nonconscious processes in individuals who are subsequently amnesic, but who were conscious at the time of the traumatic episode and through subsequent appraisal processes in individuals who were unconscious during the traumatic episode. PMID- 10467555 TI - The effects of exposure to community violence on Khmer refugee adolescents. AB - The aim of the study was to examine the relationship between exposure to violence and mental health outcomes among Khmer refugee adolescents. Seventy-six Khmer junior and senior high school students and their parents participated in a cross sectional survey study. Half of the adolescents survived violence directed at them, and two thirds witnessed violence. Although approximately one quarter either partially or fully met the criteria for posttraumatic stress disorder (PTSD), most were functioning adequately. Ordinary least squares regression analyses indicated that the number of violent events they were exposed to in their lifetime significantly predicted their level of functioning and PTSD. Recommendations for future research and implications for clinicians and educators working with Khmer and other refugee adolescents are discussed. PMID- 10467556 TI - An examination of antecedent traumas and psychiatric comorbidity among male inmates with PTSD. AB - Despite substantially higher rates of posttraumatic stress disorder (PTSD) among male inmates than among men in the general population, there is a dearth of research on PTSD among incarcerated men. The current study addresses traumatic events that precede PTSD and psychiatric disorders that are comorbid with PTSD in an inmate sample. Seeing someone seriously injured or killed, being sexually abused, and being physically assaulted were the three most commonly reported antecedent traumas to PTSD. Lifetime and current rates of mood disorders, anxiety disorders, and antisocial personality disorder were elevated among inmates with a diagnosis of PTSD. Two hundred and thirteen inmates participated in the study. Sixty-nine participants (33%) met lifetime DSM-III-R criteria for PTSD, and 45 (21%) met current criteria. The findings are compared with general population samples, and implications of the findings are discussed. PMID- 10467557 TI - Validation of the PTSD Checklist-Civilian Version in survivors of bone marrow transplantation. AB - Life-threatening illness now qualifies as a precipitating stessor for posttraumatic stress disorder (PTSD). We examined the validity of the PTSD Checklist-Civilian Version (PCL-C; Weathers, Litz, Herman, Juska, & Keane, 1993), a brief 17-item inventory of PTSD-like symptoms, in a sample of 111 adults who had undergone bone marrow transplantation an average of 4.04 years previously. Exploratory factor analysis of the PCL-C identified four distinct patterns of symptom responses: Numbing-Hyperarousal, Dreams-Memories of the Cancer Treatment, General Hyperarousal, Responses to Cancer-Related Reminders and Avoidance Numbing. Respondents meeting PTSD symptom criteria on the PCL-C had significantly lower physical, role, and social functioning, greater distress and anxiety, and significantly more intrusive and avoidant responses than individuals who did not meet PTSD symptom criteria. PMID- 10467558 TI - Analysis of violent behavior in Vietnam combat veteran psychiatric inpatients with posttraumatic stress disorder. AB - This study tested the hypothesis that male Vietnam veterans seeking inpatient treatment for PTSD (n = 228) exhibit more violent behavior compared with a mixed diagnostic group of male psychiatric inpatients without PTSD (n = 64) and a community sample of Vietnam veterans with PTSD not undergoing inpatient treatment (n = 273). Violent acts assessed included property destruction, threats without a weapon, physical fighting, and threats with a weapon. PTSD inpatients engaged in more types of violent behavior than both comparison conditions. Correlates of violence among PTSD inpatients included PTSD symptom severity and, to a lesser degree, measures of substance abuse. These findings justify routine assessment of violent behavior among inpatient with PTSD, as well as application of specialized interventions for anger dyscontrol and aggression. PMID- 10467559 TI - Predictors of acute stress following motor vehicle accidents. AB - The aim of this study was to investigate the predictors of acute stress following motor vehicle accidents (MVA). Sixty-two consecutive adult admissions to a hospital were assessed between two days and four weeks following a MVA. Participants were assessed for acute stress disorder (ASD) with a structured clinical interview and administered the Beck Depression Inventory (BDI), Coping Style Questionnaire, Dissociative Experiences Scale, and the Eysenck Personality Inventory. The prevalences of full and sub-syndromal ASD were 16.1% and 14.5%, respectively. BDI, history of psychiatric treatment, history of posttraumatic stress disorder (PTSD), and history of previous MVA accounted for 61% of the variance of acute stress severity. The findings indicate that predictors of acute stress severity are comparable to the predictors of PTSD and highlight the possibility of identifying those who may benefit from early treatment. PMID- 10467560 TI - Memory functioning in adult women traumatized by childhood sexual abuse. AB - Memory impairment has been reported in some studies of patients with combat related posttraumatic stress disorder (PTSD) and in rape victims with PTSD. The authors tested whether explicit memory impairment was evident in adult women who were traumatized by severe sexual abuse in childhood. The California Verbal Learning Test (Delis, Kramer, Kaplan, & Ober, 1987) and the Benton Visual Retention Task (Benton, 1974), were administered to 22 female adult survivors of childhood sexual trauma and to 20 demographically and educationally similar nonvictimized women. No evidence was found of explicit memory impairment in the abuse survivors. Furthermore, neither PTSD severity, dissociative symptom severity, nor extent of preexisting amnesia for childhood trauma contributed to the variance in memory functioning. Additional studies are needed to determine the extent to which impaired explicit memory functioning is a common feature of posttraumatic stress syndromes. PMID- 10467561 TI - Effects of varying scoring rules of the Clinician Administered PTSD Scale (CAPS) for the diagnosis of PTSD after acute burn injury. AB - The Clinician Administered Posttraumatic Stress Disorder Scale (CAPS) enables quantification of the severity of each of the 17 posttraumatic stress disorder (PTSD) symptoms. Blanchard and colleagues (1995) have documented variation in rates of PTSD among survivors of motor vehicle accidents depending on the CAPS scoring rule used. This report examines the effects of varying the scoring rules of the CAPS on rates of acute PTSD symptoms in hospitalized burn patients. Changing from the most liberal to the most conservative scoring rule resulted in a change in diagnosis of acute PTSD from 25% of 32% of the sample. The variation documented in this study and others has implications for a range of issues, including rates of PTSD in epidemiological studies, treatment outcome research, and forensic evaluations. PMID- 10467562 TI - Cognitive decline associated with normal aging in rats: a neuropsychological approach. AB - The effects of aging on cognitive capacities were examined by comparing the performance of young and old rats on tasks characterized as dependent on different brain systems. This neuropsychological approach was employed to determine the extent to which multiple neural systems are compromised in aging and whether deterioration of one system correlates with that of another. The two tasks used in the present study were an odor-guided recognition memory task, for which different aspects of performance have been shown to be dependent on the integrity of the orbital prefrontal and perirhinal-entorhinal cortex, and the Morris water maze, for which performance depends on the medial prefrontal cortex and hippocampus. Rats were trained on the recognition memory task under minimal memory load and then challenged with longer memory delays and higher levels of inter-item interference. Considerable variation was observed in the performance of aged rats on acquisition of the recognition memory task, and unlike young rats, some aged rats could not acquire the task. Nevertheless, those aged rats who did acquire the cDNM task performed as well as young animals when the memory delay was extended and interference was elevated. In addition, consistent with previous findings, the performance of the same aged rats was highly variable in the Morris water maze task. Furthermore, although correlations between scores on the two tasks for individual aged rats were not reliable, only those aged rats that performed outside the performance range of young rats in the water maze were impaired on acquisition of the recognition memory task. This pattern of findings is consistent with age-related dysfunction in multiple subdivisions of the prefrontal cortex as well as the hippocampus and suggests that these brain regions may deteriorate in the same subgroup of aged rats. PMID- 10467563 TI - A role for a chicken homolog of the neural cell adhesion molecule L1 in consolidation of memory for a passive avoidance task in the chick. AB - Intracranial injection of antibodies directed against the neural cell adhesion molecule L1 resulted in amnesia for passive avoidance training in day-old chicks tested 24 hr subsequently. L1 antibodies were amnesic when administered at one of two time windows: 30 min pretraining and 5.5-8 hr post-training. No amnesia was apparent if injections were made at times before, between, or after these time windows (-2, +1, +3, +4, or +12 hr relative to training). A fragment of the L1 molecule derived from the external fibronectin domains FN1-5 produced amnesia only when injected at the 5.5-hr timepoint, whereas a fragment of the immunoglubin-like domains Ig I-VI produced amnesia only when injected 30 min prior to training. We have shown previously that long-term memory for the passive avoidance task requires two waves of glycoprotein synthesis, the first occurring immediately after training, and the second some 6 hr thereafter. The glycoprotein synthesis inhibitor 2-deoxygalactose results in amnesia if injected at either time, whereas the neural cell adhesion molecule (N-CAM) is specifically involved only in the second wave. The coincidence of the time course of memory disruption resulting from injection of L1 antibodies with that occurring with 2 deoxygalactose supports the hypothesis that establishment of an enduring memory for the experience of passive avoidance training requires two waves of glycoprotein synthesis, each wave being biochemically and functionally discrete. The differential effects of the two L1 fragments suggests that separate mechanisms of synaptic stabilization are involved at the two time points. PMID- 10467564 TI - Overexpression of a calcium-binding protein, S100 beta, in astrocytes alters synaptic plasticity and impairs spatial learning in transgenic mice. AB - Recent evidence suggests that slowly propagating Ca2+ waves from astrocytes can modulate the function of neurons. Altering astrocytic calcium processes in vivo may therefore affect neuronal and behavioral phenotypes. Previously, we generated transgenic mice that overexpress an astrocytic calcium-binding protein, S100 beta. Immunocytochemistry and in situ hybridization showed elevated expression in the astrocytes of the hippocampus and other brain regions. Neurons in the hippocampus were negative for S100 beta. In this paper we analyze the hippocampal electrophysiology and learning properties of mice from two transgenic lines. Significant differences were found between the hippocampal slices of normal and transgenic mice in their response to high frequency (100 Hz) stimulation. The overall distribution of post-tetanic excitatory postsynaptic potentials (EPSP) of the slices from the transgenic mice was shifted significantly toward smaller values to a degree that 25% of slices exhibited depression. The altered hippocampal neurophysiology was accompanied by an impairment in a hippocampal dependent learning task. Transgenic mice showed significant impairment in a spatial version of the Morris water maze, however, they performed normally in non spatial tasks. Probe trials showed that transgenic mice, though significantly impaired, also acquired spatial information. The results suggested that the impairment was not due to motor dysfunction, impaired vision or motivation of the transgenic mice, findings compatible with a possible hippocampal mechanism. We conclude that overexpression of S100 beta in astrocytes impairs, but does not abolish, the ability to solve a spatial task, and it leads to a significantly decreased post-tetanic potentiation in the hippocampal slice. We hypothesize that the changes are due to calcium mediated processes. Our results support the notion that astrocytes are involved in higher brain functions. PMID- 10467565 TI - Environmental enrichment and postnatal handling prevent spatial learning deficits in aged hypoemotional (Roman high-avoidance) and hyperemotional (Roman low avoidance) rats. AB - The present study investigated the enduring effects of postnatal handling (administered during the first 21 days of life), and environmental enrichment (for a period of 6 months starting 3 weeks after weaning) on spatial learning in 24-month-old hypoemotional (Roman high-avoidance, RHA/Verh) and hyperemotional (Roman low-avoidance, RLA/Verh) rats. Two groups of 5-month-old rats from both lines were also included in the experiment as young controls. The Roman lines performed differently in the Morris water maze: Path lengths of RLA/Verh rats were shorter and they swam at lower speed than RHA/Verh rats, showing quicker and more efficient learning overall. Postnatal handling improved learning mainly in RHA/Verh rats, whereas environmental enrichment was able to prevent the deficits shown by aged controls of both lines. Young, enriched, and handled plus enriched animals exhibited better performance than impaired aged controls, to the point that aged enriched and handled plus enriched animals did not differ from young controls. Thus, besides indicating that RLA/Verh rats are better learners than RHA/Verh rats in the Morris water maze, this study demonstrates that environmental enrichment prevents the cognitive loss associated with aging, over the long term. Finally, the positive effects obtained with postnatal handling were dependent on the rat line. PMID- 10467566 TI - Analysis of conditioned courtship in dusky-Andante rhythm mutants of Drosophila. AB - Genetic connections between learning and rhythmicity were suggested to have been established in a previous study, in part because the duskyAndante (dyAnd) mutation in Drosophila disrupted both behaviors. dyAnd, isolated as a slow-clock variant, was reported to cause an approximately fourfold decrement in courtship suppression conditioning. These effects have been reexamined; the experiments were buttressed by testing the effects of several recently isolated mutations at the dusky locus, along with the original And Allele that had been induced there. The reexamination was also prompted by anatomical concerns, certain of which have recently focused on dy-induced decrements in cell size, but only in terms of wing morphology. Another anatomical issue involves the discovery of a neuronal pathway that seems to connect circadian pacemaker cells to a structure in the Drosophila brain that is involved in learning. In observer-blind experiments, however, it was found that neither pacemaker-slowing (Andante-like) dy mutations nor others that cause no rhythm defects produced subnormal conditioned courtship. Moreover, in the adult brain of a slow-clock dyAnd mutant, no axonal pathway defects were readily discernible and putative pacemaker neurons appeared to be normal in size. PMID- 10467567 TI - Associative learning and stimulus novelty influence the song-induced expression of an immediate early gene in the canary forebrain. AB - To identify variables that affect immediate early gene (IEG) expression in the auditory telencephalon of songbirds, we developed a conditioning paradigm that trained adult male canaries to associate song with a mild shock. Learning of the association was measured by a bird's fear and avoidance responses. Birds exposed to paired song and shock were compared to yoked controls exposed to each stimulus alone or to both unpaired. Additional groups examined the effects of attention and stress, and of the novelty of the stimulus situation. In situ hybridization analysis of brain sections revealed an enhancement of ZENK expression in birds learning the association between song and shock above levels induced by song alone or yoked-unpaired song and shock. This effect was specifically seen in the caudomedial auditory telencephalon (NCM-HVCM). A comparison of the several control groups indicated that novelty of the song stimulus or of its pairing with shock were the main variables that predicted ZENK levels in NCM-HVCM. These observations are compatible with ZENK playing a role in the formation of song perceptual memories. PMID- 10467568 TI - A critical period of protein kinase activity after tetanic stimulation is required for the induction of long-term potentiation. AB - A critical period of protein kinase activity required for the induction of long term potentiation (LTP) was determined in area CA1 or hippocampal slices using the broad-range and potent protein kinase inhibitors K-252a and staurosporine. As reported previously, K-252a and staurosporine blocked LTP induction when applied before, during, and after high-frequency stimulation (HFS). In contrast, K-252a did not block LTP when applied only before and during HFS and washed out immediately after HFS. K-252a and staurosporine both attenuated LTP magnitude when applied immediately after or as late as 5 min after HFS. However, K-252a applications beginning 30-45 min after HFS did not affect LTP expression significantly. K-252a had no detectable effect on isolated N-methyl-D-aspartate (NMDA) receptor-mediated EPSPs but significantly inhibited the in situ phosphorylation of specific hippocampal proteins (synapsin I, MARCKS, and B-50). In addition, K-252a attenuated 4 beta-phorbol-12,13-dibutyrate (PDBu)-enhanced synaptic transmission. Our results indicate that there is a critical period of protein kinase activity required for LTP induction that extends for approximately 20 min after HFS. In addition, our results suggest that protein kinase activity during and immediately after HFS is not sufficient for LTP induction. These results provide new information about the mechanisms that underlie LTP induction and expression and provide evidence for persistent and/or Ca(2+)-independent protein kinase activity involvement in LTP. PMID- 10467569 TI - Long-term potentiation and evidence for novel synaptic association in CA1 stratum oriens of rat hippocampus. AB - In CA1 stratum oriens of hippocampal slices, a robust long-term potentiation (LTP) induced by tetanic stimulation (20 pulses at 100 Hz, 10 trains delivered at 0.1 Hz) was accompanied by a slowly developing potentiation in a second, untetanized pathway. N-methyl-D-aspartate (NMDA) receptor antagonist DL-2-amino-5 phosphonovaleric acid (D-APV, 50 or 100 microM) reduced the homosynaptic LTP by 80% but failed to affect heterosynaptic LTP. The metabotropic receptor antagonist DL-2-amino-3-phosphonopropionic acid DL-AP3, 300 microM) or (+)-alpha-methyl-4 carboxyphenylglycine (MCPG, 500 microM), applied with DL-APV, further reduced homosynaptic LTP and blocked heterosynaptic LTP. The inhibitor of Ca(2+)-induced Ca2+ release dantrolene (20 microM), also applied with DL-APV, blocked both components of LTP. Importantly, when low-frequency test stimulation (0.017 Hz) to the untetanized, heterosynaptic pathway was interrupted for 30 min after homosynaptic tetanization, heterosynaptic LTP did not develop. These results demonstrate homosynaptic and heterosynaptic LTP inductions in stratum oriens of the area CA1 and suggest that the heterosynaptic LTP is induced by NMDA independent, novel associative processes between tetanized and untetanized pathways. PMID- 10467570 TI - Remembering and forgetting as context discrimination. AB - In 1929, H.C. Blodgett reported the results of a seminal maze learning experiment using rats. In that experiment, hungry rats ran in a complex maze but were not rewarded on reaching the goal box. Not surprisingly, the performance of the hungry rats did not improve over trials. However, with the introduction of reward, the error scores of the rats suddenly dropped to the level of the control rats that were rewarded from the outset. This finding indicates that the experimental group had learned the maze despite the absence of reward but that the learning was latent rather than manifest. With Blodgett's findings, the distinction between learning and performance became firmly established, if not as widely appreciated as it might be. Blodgett's (1929) early experimental finding of latent learning could well serve as a paradigm for the approach taken here. That is, we have emphasized the principle that a lack of performance does not necessarily indicate a lack of either learning or memory. This principle is much more than an empty admonition: We have shown it can have a firm theoretical basis, one that has been confirmed repeatedly by experiments cited throughout this paper. That is, it has been shown numerous times that a failure to perform either in a Pavlovian or instrumental learning task or to remember in an animal or human memory task under one set of conditions could be alleviated under another set of conditions. Forgetting was viewed here as a failure of performance resulting from the cues at test retrieving a memory other than the target memory or retrieving no memory at all. According to this view, memory involves discrimination learning. Essentially, memories are stored in the presence of an elaborate set of interoceptive and exteroceptive stimuli, a context. Whether at test the target memory is retrieved depends on how well the cues at test discriminate between the target memory and other memories. This approach suggests that forgetting does not occur: There is only a failure to perform because of a difference between the stimulus conditions prevailing at encoding and at test. It was demonstrated that this approach is at least as reasonable as that which suggests that true forgetting occurs, but certainly more useful. At least three advantages adhere to our view that memory is a discrimination problem. First, in almost numberless cases, it has been shown that failure of performance under one set of stimulus conditions can be alleviated under some other set of stimulus conditions. Second, the proposition that altered stimulus conditions are responsible for forgetting is one of wide generality. Thus, the altered stimulus conditions approach can serve as an explanation not only for various human memory findings but also for various animal memory and learning findings. Finally, and perhaps most importantly, the present approach provides investigators with a powerful and proven working hypothesis. It tells us not to accept failures of performance as indicating an absence of learning or a loss of memory but rather to seek conditions favorable to improving performance, a strategy that should lead to a better fundamental understanding of memory and learning. This position is obviously a type of optimality theory, of which evolutionary theory is one of the more outstanding examples. In optimality theory, any deviation from some ideal state or condition prompts the investigator to seek the reasons for deviation. This approach may prove as successful when applied to learning and memory as it has to other areas of science. PMID- 10467571 TI - Role of interneurons in defensive withdrawal reflexes in Aplysia. PMID- 10467572 TI - Behavioral analysis of Drosophila landmark learning in the flight simulator. AB - Drosophila flies can be trained in the flight simulator to operantly avoid heat by choosing certain orientations relative to landmarks. Flies primarily store pattern orientations associated with the absence of heat. They readily escape from heat-associated orientations under the direct influence of the reinforcer but not in the subsequent memory tests. The flies tend to keep the largest possible distance from the "hot" or potentially "hot" regions, that is, they head toward the center of the "cold" sector. The results are discussed in the light of the retinotopic matching model explaining visual memory in flies by the superposition of a retinotopically stored template with the actual retinal image. Window experiments confining visual feedback to two 90 degrees sectors indicate that the memory template covers most of the visible space. PMID- 10467573 TI - Heat shock disrupts long-term memory consolidation in Caenorhabditis elegans. AB - Previous work has demonstrated that memory for habituation training is retained for > 24 hr in Caenorhabditis elegans. In this study the timing of memory consolidation was investigated by introducing heat shock (32 degrees C, 45 min) either before training, long after training, or during training. It was found that memory consolidation was disrupted by heat shock during training but not before or after training. In addition, heat shock before training failed to induce thermal tolerance to the effects of heat shock during training on long term memory formation. When brief heat shock (32 degrees C, 15 min) was presented during training at different intervals, the results suggested that a narrow critical period for memory consolidation of habituation may exist. These findings demonstrate that in C. elegans long-term memory for habituation is disrupted by a temporally defined agent, heat shock. Therefore, heat shock can be used as a fine grained tool to investigate the dynamics of memory consolidation. PMID- 10467574 TI - Direct comparison of serotonin effects on siphon versus tail sensory neurons in Aplysia. AB - Modulation of the strength of siphon and tail withdrawal reflexes in Aplysia involves, in part, changes in the sensory neurons that initiate these reflexes. Different observations and experiments on modulation in siphon and tail sensory neurons together contribute to the working model of mechanisms of learning and memory in Aplysia, yet no direct comparison of modulation in these two classes of sensory neurons has yet been made. The purpose of the present study was to directly compare modulation between siphon and tail sensory neurons in the same experimental conditions. In particular, we focused on the effects of serotonin on two firing properties of sensory neurons: spike duration and excitability. We applied serotonin (5-HT) onto both siphon and tail sensory neurons under the same conditions and found that both spike duration and excitability were significantly enhanced. This enhancement was statistically indistinguishable between siphon and tail sensory neurons tested simultaneously in the same preparation. Thus, these two different classes of sensory neurons respond to 5-HT in very similar, if not identical, ways. We conclude that if there are differences in 5-HT induced modulation between siphon and tail sensory neurons at the biophysical level, such differences are not strongly manifested at the level of changes in firing properties. PMID- 10467576 TI - Behavioral development in the honey bee: toward the study of learning under natural conditions. PMID- 10467575 TI - Temporal specificity of long-term depression in parallel fiber--Purkinje synapses in rat cerebellar slice. AB - The phenomenon of cerebellar long-term depression (LTD), a decrease of synaptic strength between the parallel fibers (PFs) and Purkinje cells after conjunctive activation of PFs and the climbing fibers (CFs), is implicated as a cellular mechanism for motor learning. We have characterized a field-potential recording technique in cerebellar slice and have used the technique to examine the temporal conditions for cerebellar LTD induction in an attempt to examine the relevance of LTD to associative conditioning. Interstimulus intervals (ISIs) between onsets of PF and CF activation and the number of paired stimuli (pairings) were examined. LTD has distinct temporal specificity that seems to be constrained by inhibitory interneurons and can be masked by excessive stimulation. When 100 paired stimuli were given to PFs and CFs, LTD was induced with an ISI of 250 msec (PF activation preceding CF activation). In contrast, a smaller forward (125 msec), simultaneous (0 msec), or backward (-250 msec) ISIs were not effective for inducing LTD. However, the blockade of GABAA receptor-mediated inhibition made it possible to induce LTD with 100 pairs of simultaneous stimulations. Furthermore, by increasing the number of pairings to 600, significant LTD was observed with all four interstimulus intervals. These results suggest that temporal conditions for LTD induction share some similarity to associative learning of discrete motor responses. PMID- 10467577 TI - Learning "what" and "how" in a human motor task. AB - We studied the development of implicit and of verbally declared knowledge for normal human subjects who learned an unfamiliar motor task in one learning session. The exploratory nature of motor learning and a special period for optimizing skill were followed in real time. Subjects understood the goal for task success, but they had to learn a motor strategy of what pattern of serial movements to make and the tactics of how much to scale their amplitudes and timing. We compared the time course for acquiring tactical skill with that for acquiring knowledge of strategy and of tactics, and their necessary cues. Implicit and declarative knowledge were distinguished from one another by correlating subjects' verbal self-reports with movement kinematics and their results. Implicit generation of the correct strategy and of the tactics developed in an exploratory manner from the beginning of the learning session. Implicit strategy learning soon gave way to conscious efforts, but tactical learning remained implicit until its first unambiguous verbal declaration (with one exception). First strategy declarations were voiced before those for tactics, during trial-and-error learning that did not require task success, and referred to reversing the direction of hand movements (one exception). In contrast, first declarations of tactics almost always required actual or imminent success, referred to when direction was to be reversed, and it was achieved near the top of a sigmoid learning curve that rose to tactical skill (with one exception). During the sigmoid rise, movement amplitudes and timing were optimized in a distinct manner, although tactics usually adapted thereafter to movements of more moderate speed that could still be successful. PMID- 10467578 TI - Metabotropic glutamate receptors in spatial and nonspatial learning in rats studied by means of agonist and antagonist application. AB - We examined the effects of both the metabotropic glutamate receptor (mGluR) antagonist MCPG and the agonist tADA in two behavioral paradigms in rats: (1) brightness discrimination and (2) spatial alternation. Compounds were applied intracerebroventricularly at different times, either 30 min prior to training or immediately after training, and rats were tested for retention 24 hr later in the same paradigms. Both MCPG and tADA caused amnesia in the spatial alternation test, when applied pretraining, but no effect was obtained in the brightness discrimination paradigm. Drug-induced amnesia was shown not to be attributable to state-dependent effects of MCPG or tADA. Moreover, the memory inhibiting effect of MCPG was dose dependent, with a low dose (20 mM/5 ml) having no effect on learning and memory and a 10 times higher concentration (200 mM/5 ml) causing complete amnesia. Application of both saline and MCPG immediately post-training prevented memory formation, which may be attributable to an interference by the injection procedure with the process of memory formation. The mGluR agonist tADA, however, facilitated memory formation in the spatial alternation task, when injected immediately after training. Post-training application of the compounds had no effect on retention in the brightness discrimination task. On the basis of these data we conclude that (1) mGluRs are of particular importance for spatial learning and play no role in visual discrimination; (2) both the block and the activation of mGluRs inhibit spatial learning, suggesting that saturated activation prevents further modulation of mGluRs, which may be required during learning or memory formation; and (3) mGluR agonist tADA may be memory facilitating when applied after training, thus enhancing the establishment of the memory trace. PMID- 10467579 TI - 7-Nitro indazole, a selective neuronal nitric oxide synthase inhibitor in vivo, impairs spatial learning in the rat. AB - Nitric oxide (NO) is an intercellular messenger that has been suggested to have a role in learning and memory formation. Previous studies with nonselective NO synthase inhibitors have produced contradictory results in learning experiments. However, these drugs also produced blood pressure changes, as NO is an endothelial-derived relaxing factor. A novel NO synthase inhibitor, 7-nitro indazole (7-NI), as a dose (30 mg/kg i.p.) shown previously to inhibit neuronal NO synthase by 85% without affecting blood pressure, produced amnesic effects both in a water maze and in an 8-arm radial maze. Latency as well as distance was greater in the 7-NI group in the water maze while swim speed was not affected. Latency, working memory (WM), and reference memory (RF) errors were also higher in the 7-NI group in the 8-arm maze. At the end of the second training day, these differences were no longer apparent. However, on the fourth training day, a transfer test in the water maze showed that 7-NI had produced a spatial memory deficit, reducing quadrant bias and the number of annulus crossings. Learning of a visual cue task was not affected. No difference between groups was visible in an open field test. We conclude that neuronal NO synthase activity plays a role in learning and memory formation in the rat. PMID- 10467580 TI - Higher-order associative processing in Hermissenda suggests multiple sites of neuronal modulation. AB - Two important features of modern accounts of associative learning are (1) the capacity for contextual stimuli to serve as a signal for an unconditioned stimulus (US) and (2) the capacity for a previously conditioned (excitatory) stimulus to "block" learning about a redundant stimulus when both stimuli serve as a signal for the same US. Here, we examined the process of blocking, thought by some to reflect a cognitive aspect of classical conditioning, and its underlying mechanisms in the marine mollusc Hermissenda. In two behavioral experiments, a context defined by chemosensory stimuli was made excitatory by presenting unsignalled USs (rotation) in that context. The excitatory context subsequently blocked overt learning about a discrete conditioned stimulus (CS; light) paired with the US in that context. In a third experiment, the excitability of the B photoreceptors in the Hermissenda eye, which typically increases following light-rotation pairings, was examined in behaviorally blocked animals, as well as in animals that had acquired a normal CS-US association or animals that had been exposed to the CS and US unpaired. Both the behaviorally blocked and the "normal" learning groups exhibited increases in neuronal excitability relative to unpaired animals. However, light-induced multiunit activity in pedal nerves was suppressed following normal conditioning but not in blocked or unpaired control animals, suggesting that the expression of blocking is mediated by neuronal modifications not directly reflected in B-cell excitability, possibly within an extensive network of central light-responsive interneurons. PMID- 10467581 TI - Integration of olfactory information in a spatial representation enabling accurate arm choice in the radial arm maze. AB - The aim of the present study was to determine whether and how rats can use local olfactory cues for spatial orientation. Rats were trained in an eight-arm radial maze under different conditions as defined by the presence or absence of supplementary olfactory cues marking each arm, the availability of distant visuospatial information, and the illumination of the maze (light or darkness). The different visual conditions were designed to dissociate among the effects of light per se and those of visuospatial cues, on the use of olfactory cues for accurate arm choice. Different procedures with modifications of the arrangement of olfactory cues were used to determine if rats formed a representation of the spatial configuration of the olfactory cues and if they could rely on such a representation for accurate arm choice in the radial maze. The present study demonstrated that the use of olfactory cues to direct arm choice in the radial arm maze was critically dependent on the illumination conditions and implied two different modes of processing of olfactory information according to the presence or the absence of light. Olfactory cues were used in an explicit manner and enabled accurate arm choice only in the absence of light. Rats, however, had an implicit memory of the location of the olfactory cues and formed a representation of the spatial position of these cues, whatever the lighting conditions. They did not memorize the spatial configuration of the olfactory cues per se but needed these cues to be linked to the external spatial frame of reference. PMID- 10467582 TI - Effects of a conditional Drosophila PKA mutant on olfactory learning and memory. AB - The requirement for cAMP-dependent protein kinase (PKA) in associative learning of Drosophila was assessed in mutant flies hemizygous for a cold-sensitive allele, X4, of the DC0 gene, which encodes the major catalytic subunit of PKA. DC0X4 hemizygotes died as third-instar larvae at 18 degrees C, the restrictive temperature, but were viable when raised at 25 degrees C. Shifting adult DC0X4 hemizygotes from 25 degrees C to 18 degrees C led to a decrease in PKA activity from 24% to 16% of wild-type without impairing viability. At 25 degrees C, DC0X4 hemizygotes exhibited reduced initial learning relative to controls but normal memory decay in a Pavlovian olfactory learning assay. Shifting the temperature from 25 degrees C to 18 degrees C prior to training reduced initial learning to a similar extent in DC0X4 hemizygotes and controls but resulted in a steeper memory decay curve only in DC0X4 hemizygotes. These observations are suggestive of a role for PKA in medium-term memory formation in addition to its previously established role in initial learning. PMID- 10467583 TI - The predictive brain: temporal coincidence and temporal order in synaptic learning mechanisms. AB - Some forms of synaptic plasticity depend on the temporal coincidence of presynaptic activity and postsynaptic response. This requirement is consistent with the Hebbian, or correlational, type of learning rule used in many neural network models. Recent evidence suggests that synaptic plasticity may depend in part on the production of a membrane permeant-diffusible signal so that spatial volume may also be involved in correlational learning rules. This latter form of synaptic change has been called volume learning. In both Hebbian and volume learning rules, interaction among synaptic inputs depends on the degree of coincidence of the inputs and is otherwise insensitive to their exact temporal order. Conditioning experiments and psychophysical studies have shown, however, that most animals are highly sensitive to the temporal order of the sensory inputs. Although these experiments assay the behavior of the entire animal or perceptual system, they raise the possibility that nervous systems may be sensitive to temporally ordered events at many spatial and temporal scales. We suggest here the existence of a new class of learning rule, called a predictive Hebbian learning rule, that is sensitive to the temporal ordering of synaptic inputs. We show how this predictive learning rule could act at single synaptic connections and through diffuse neuromodulatory systems. PMID- 10467584 TI - Lesions of the dorsal hippocampal formation interfere with background but not foreground contextual fear conditioning. AB - The effects of hippocampal lesions on the conditioning of fear responses (freezing responses) to contextual stimuli (static, continuously present stimuli) were examined in three conditioning paradigms: forward pairing of a phasic tone conditioned stimulus (CS) with a footshock unconditioned stimulus (US), unpaired presentations of the CS and US, or presentations of the US alone. All three procedures resulted in the acquisition of conditioned freezing to contextual stimuli. Lesions of the dorsal hippocampus prevented the acquisition of contextual conditioning in the Paired procedure, as reported previously, but not in the Unpaired or US Alone procedures. In the Paired procedure, static contextual cues occur in the background, with the phasic tone CS being the primary stimulus that enters into the association with the US. However, in the other two procedures, where there is no phasic CS, the primary associations with the US involve static contextual stimuli, which are therefore in the foreground. We refer to these types of contextual conditioning as background and foreground contextual conditioning, respectively, and argue that the hippocampus is only involved in background contextual conditioning. These results have implications for understanding both fear conditioning and hippocampal function. PMID- 10467585 TI - Hippocampal formation size in normal human aging: a correlate of delayed secondary memory performance. AB - Although mild progressive memory impairment is commonly associated with normal human aging, it is unclear whether this phenomenon can be explained by specific structural brain changes. In a research sample of 54 medically healthy and cognitively normal elderly persons (ages 55-87, x = 69.0 +/- 7.9), magnetic resonance imaging (MRI) was used to derive head-size-adjusted measurements of the hippocampal formation (HF) (dentate gyrus, hippocampus proper, alveus, fimbria, subiculum), the superior temporal gyrus (STG), and the subarachnoid cerebrospinal fluid (CSF) (to estimate generalized cerebral atrophy). Subjects were administered tests of primary memory (digit span) and tests of secondary memory with immediate and delayed recall components (paragraph, paired associate, list recall; facial recognition). Separate composite scores for the immediate and delayed components were created by combining, with equal weighting, the subtests of each category. The WAIS vocabulary subtest was used as a control measure for language and intelligence. A highly significant correlation (P < 0.001), independent of age, gender, and generalized cerebral atrophy was found between HF size and delayed memory performance. No significant correlations were found between HF size and primary or immediate memory performance. STG size was not significantly correlated with any of the composite memory variables. These results suggest that HF atrophy may play an important independent role in contributing to the memory loss experienced by many aging adults. PMID- 10467586 TI - Potentiation of dentate synapses initiated by exploratory learning in rats: dissociation from brain temperature, motor activity, and arousal. AB - Certain kinds of learning may be related to potentiation of transmission at specific hippocampal synapses. We investigated whether transmission across the perforant-path/granule-cell synapses of the dentate gyrus is facilitated when rats are learning about novel objects in an open field during exploration. Such studies are complicated by the sensitivity of hippocampal field potentials to brain temperature change. To control for this, we have recorded both brain temperature and field potentials and compared potentials sampled during exploration with potentials taken at corresponding brain temperature in a passive warming situation, with the animals at rest. Relative to these reference potentials, both the f-EPSP slope and the population spike were elevated while the rats explored. The potentiation reached its maximum within < 5 sec after the exploration began. During the first 2 min, the f-EPSP slope was enhanced by 6.5% relative to the control values. The potentiation then decayed, reaching the reference values after 20-30 min of exploration. Significant potentiation required exploration above a certain minimum intensity. Control experiments showed that the changes were neither mimicked by arousal in response to aversive stimuli nor by motor activity. It is suggested that the facilitated transmission across the perforant-path/dentate synapses may be involved in learning during exploration. PMID- 10467587 TI - Recruitment of long-lasting and protein kinase A-dependent long-term potentiation in the CA1 region of hippocampus requires repeated tetanization. AB - To study how the late phase of long-term potentiation (LTP) in hippocampus arises, we examined the resulting LTP for its time course and its dependence on protein synthesis and different second-messenger kinases by applying various conditioning tetani. We find that one high-frequency train (100 Hz) produces a form of LTP that lasts longer than 1 hr but less than 3 hr (the early phase of LTP, or E-LTP). It is blocked by inhibitors of calcium/calmodulin kinase II (Cam kinase II) but is not affected by an inhibitor of cAMP-dependent protein kinase [protein kinase A (PKA) and the protein synthesis inhibitor anisomycin] nor is it occluded by the cAMP activator forskolin. In contrast, when three high-frequency trains are used, the resulting potentiation persists for at least 6-10 hr. The L LTP induced by three trains differs from the E-LTP in that it requires new protein synthesis, is blocked by an inhibitor of cAMP-dependent protein kinase, and is occluded by forskolin. These results indicate that the two mechanistically distinctive forms of LTP, a transient, early component (E-LTP) and a more enduring form (L-LTP), can be recruited selectively by changing the number of conditioning tetanic trains. Repeated tetani induce a PKA and protein synthesis dependent late component that adds to the amplitude and duration of the potentiation induced by a single tetanus. PMID- 10467588 TI - Neuronal tuning and associative mechanisms in form representation. AB - We examine the hypothesis that the form representation in the anterior inferotemporal (AIT) cortex is acquired through learning. According to this hypothesis, perceptual aspects of the temporal association area are closely related to its visual representation, in that the response selectivity of AIT neurons can be influenced by visual experience. On the basis of the neurophysiological evidence, we summarize two neuronal mechanisms that subserve the acquisition of form selectivity in AIT neurons. The first mechanism is neuronal tuning to particular stimuli that were learned in a cognitive task. The second mechanism is association, with which relevant information can be retrieved from other stored memories. On the grounds that long-term memory of objects is acquired and organized by at least these two neuronal mechanisms in the temporal association area, we further present a model of the cognitive memory system that unifies perception and imagery. PMID- 10467589 TI - Probabilistic classification learning in amnesia. AB - Amnesic patients and control subjects participated in a study of probabilistic classification learning. In each of three tasks, four different cues were each probabilistically associated with one of two outcomes. On each trial, the cues could appear alone or in combination with other cues and subjects selected the outcome they thought was correct. Feedback was provided after each trial. In each task, the amnesic patients learned gradually to associate the cues with the appropriate outcome at the same rate as control subjects, improving from 50% correct to approximately 65% correct. Presumably because the cue-outcome associations were probabilistic, declarative memory for the outcomes of specific trials was not as useful for performance as the information gradually accrued across trials. Nevertheless, declarative memory does appear to make a contribution to performance when training is extended beyond approximately 50 trials, because with further training control subjects eventually outperformed the amnesic patients. It was also demonstrated that performance on the probabilistic classification task was not the result of holding knowledge of cue outcome associations in short-term memory, because both control subjects and amnesic patients demonstrated significant savings when testing was interrupted by a 5-min delay (experiment 2). Probabilistic classification learning appears to provide an analog in human subjects for the habit learning tasks that can be acquired normally by animals with hippocampal lesions. PMID- 10467590 TI - Input-specific induction of cerebellar long-term depression does not require presynaptic alteration. AB - In cerebellar long-term depression (LTD), conjunctive stimulation of parallel fiber (PF) and climbing fiber (CF) inputs to a Purkinje neuron result in a selective depression of PF-Purkinje neuron synaptic strength. This system is attractive for the study of neuronal information storage, both because of its duration and because it demonstrates input specificity. The mechanisms underlying input specificity in this system are not known, but they could involve presynaptic alterations, postsynaptic alterations, or some combination of both. To allow for an unambiguous analysis of postsynaptic processes, an LTD induction protocol has been developed using cultured cerebellar cells in which pulses of quisqualate and direct Purkinje neuron depolarization replace PF and CF stimulation, respectively. Input specificity is retained in this reduced system. When multiple, nonoverlapping quisqualate application sites are used, LTD is confined to those sites that are stimulated during depolarization. This property of LTD induction is also preserved under conditions where both spontaneous and evoked neurotransmitter releases are reduced or eliminated, indicating that postsynaptic alterations are sufficient to confer input specificity. Input specific LTD may also be induced by local application of a protein kinase C (PKC) activator (1-oleoyl-2-acetylglycerol) together with direct Purkinje neuron depolarization, suggesting that input-specific LTD results from the conjunction of a spatially broad Ca signal mediated by Purkinje neuron depolarization, together with a spatially constrained PKC-activating signal mediated by quisqualate application. PMID- 10467591 TI - Low-frequency stimulation erases LTP through an NMDA receptor-mediated activation of protein phosphatases. AB - In the CA1 region of adult guinea pig hippocampal slices, long trains of theta frequency (5 Hz) stimulation produced a small enhancement of basal synaptic transmission but depressed the strength of synaptic transmission at synapses that had recently undergone long-term potentiation (LTP). Five hertz stimulation delivered immediately prior to high-frequency stimulation also inhibited the subsequent induction of LTP. The depression of potentiated synapses by 5 Hz stimulation (depotentiation) was blocked by 2-amino-5-phosphonovalerate and was observed only during the early phases of LTP. Furthermore, the protein phosphatase inhibitors okadaic acid and calyculin A blocked both depotentiation and the ability of 5 Hz stimulation to inhibit subsequent LTP, suggesting that protein phosphatases are involved in the ability of 5 Hz stimulation to modulate synaptic plasticity in the CA1 region of the hippocampus. PMID- 10467592 TI - Egr3/Pilot, a zinc finger transcription factor, is rapidly regulated by activity in brain neurons and colocalizes with Egr1/zif268. AB - Programs of gene activation may underlie long-term adaptive cellular responses to extracellular ligands. We have used a differential cDNA cloning strategy to identify genes that are strongly induced by excitatory stimuli in the adult rat hippocampus. Here, we report the rat cDNA sequence of a zinc-finger transcription factor, Egr3/Pilot, and characterize its regulated mRNA expression in brain. Egr3 mRNA is rapidly and transiently induced in neurons of the hippocampus and cortex by electroconvulsive seizure. mRNA levels peak 2 hr after the seizure and remain elevated for as long as 8 hr. Egr3 mRNA is also rapidly induced in granule cells of the dentate gyrus by synaptic NMDA receptor activation elicited by patterned stimulation of the perforant pathway and by drugs that alter dopamine neurotransmission in the striatum. Basal levels of Egr3 mRNA in the cortex appear to be driven by natural synaptic activity because monocular deprivation rapidly decreases Egr3 mRNA in the deafferented visual cortex. Aspects of the protein structure, sequence-specific DNA binding, transcriptional activity, and regulation of Egr3 are highly similar to another zinc-finger transcription factor, Egr1/zif268. Moreover, we demonstrate colocalization of Egr3 and zif268 mRNAs in neurons of normal and stimulated cortex. Our studies suggest that interactions between these coregulated transcription factors may be important in defining long-term, neuroplastic responses. PMID- 10467593 TI - CCK-A and CCK-B receptors enhance olfactory recognition via distinct neuronal pathways. AB - We have previously reported that CCK-A receptor agonists and CCK-B receptor antagonists both enhance memory in an olfactory recognition test. Here, we report that the memory-enhancing effect of the CCK-B receptor antagonist L-365,260 (1 mg/kg i.p.), but not that of the CCK-A receptor agonist caerulein (0.03 mg/kg i.p.), was dramatically decreased following a bilateral transection of the perforant path, a principal source of input to the hippocampal formation. We further confirmed that a significant memory deficit occurred subsequent to this deafferentation of the hippocampus in untreated animals. In contrast, the effect of caerulein, but not that of L-365,260, was abolished following a bilateral subdiaphragmatic vagotomy. These results demonstrate that the hippocampal system plays a role in olfactory recognition and indicate that distinct neuronal pathways underlie the memory-enhancing effects of CCK-A and CCK-B drugs observed in the olfactory recognition test. The former effects (CCK-A) appear to involve a peripheral relay to the brain via the vagus nerve, whereas the latter (CCK-B) are directly central and involve, at least in part, the hippocampal system. PMID- 10467594 TI - Sensorimotor skill learning in amnesia: additional evidence for the neural basis of nondeclarative memory. AB - We investigated sensorimotor skill learning, a form of nondeclarative (implicit) memory, in 28 subjects with declarative (explicit) memory defects caused by either mesial temporal (n = 15) or basal forebrain (n = 13) damage and in 66 normal control subjects. All 28 amnesics had normal learning of a rotor pursuit task. We also studied in detail the sensorimotor skill learning of patient Boswell. As a result of bilateral damage to both mesial and lateral aspects of the temporal lobes and to the basal forebrain, Boswell has one of the most severe impairments ever reported for learning of all types of declarative knowledge. Compared to matched controls, Boswell acquired and retained normally the skills associated with performing motor tasks. We conducted a long-term (2-year) followup study of Boswell's retention of the rotor pursuit task, and we found that he retained the skill as well as normal controls. Our study builds on previous work in the following respects: (1) It provides evidence, for the first time, that skill learning is normal in basal forebrain amnesics; (2) it shows that patient Boswell has normal learning and long-term retention of sensorimotor skills, in spite of his extensive damage; and (3) it offers additional evidence that mesial temporal lobe damage spares skill learning. These findings demonstrate unequivocally that sensorimotor skill learning does not require structures in mesial and lateral temporal regions nor in basal forebrain. PMID- 10467595 TI - Temporal and spatial regulation of the expression of BAD2, a MAP kinase phosphatase, during seizure, kindling, and long-term potentiation. AB - Recent studies indicate that stimulation of NMDA receptors in cultured hippocampal cells activates MAP kinase. Although the pathway whereby MAP kinase is activated has been been characterized, little is known about the mechanisms that shut off MAP kinase. In the course of analyzing several immediate-early genes identified previously by differential screen as inducible by seizure activity, we found that one of them, BAD2, encodes dual purpose, threonine/tyrosine phosphates with specific activity directed against MAP kinase (MKP-1). In situ hybridization of BAD2 demonstrates that stimuli that produce seizure, kindling, and long-term potentiation cause a rapid increase in BAD2 mRNA (within 0.5-1 hr after stimulation) that has, in each case, a distinctive pattern of expression in the brain. In these regions, the induction of a MAP kinase specific phosphatase may provide a negative feedback control associated with long term synaptic changes. PMID- 10467596 TI - Persistent activation of protein kinase C during the development of long-term facilitation in Aplysia. AB - We investigated activation of the two major neuronal protein kinase C (PKC) isoforms in Aplysia, Ca(2+)-activated Apl I and Ca(2+)-independent Apl II, during the induction and maintenance of behavioral sensitization of Aplysia defensive reflexes. Activation of PKC occurred during the training stimulus and persisted for at least 2 hr thereafter but was not maintained for 24 hr. The persistent activation required protein synthesis and was blocked by cyproheptidine, an agent that also blocked the initial activation of PKC. Persistent activation involved both an increase in membrane-associated Apl I and an increase in an autonomous kinase activity that may be related to a post-translational modification of Apl II. These results are consistent with the hypothesis that in addition to its role in producing the presynaptic facilitation of mechanosensory-motor neuron synapses that underlie short-term facilitation, PKC is needed for maintaining synaptic changes in an intermediate period that precedes the modifications accompanying consolidation of memory. PMID- 10467597 TI - The Atlantis platform: a new design and further developments of Buresova's on demand platform for the water maze. PMID- 10467598 TI - 7-Nitro indazole, a neuron-specific nitric oxide synthase inhibitor, produces amnesia in the chick. AB - 7-Nitro indazole (7-NI), which is selective for the neuronal isoform of nitric oxide synthase (NOS), was tested in a passive avoidance task in the chick. Injection of 50 mg/kg i.p. pretraining had amnesic effects for the task when tested 30 min, 2 or 24 hr after training. Injections post-training had no effect. Because 7-NI does not inhibit the endothelial isoform of NOS, it does not affect blood vessel relaxation, as nonspecific inhibitors do. This effect on blood vessels could explain the amnestic effects produced by nonspecific NOS inhibitors. The results support the theory that NO is a neuronal transmitter that is important in processes of synaptic plasticity and learning. PMID- 10467599 TI - Parallel brain systems for learning with and without awareness. AB - A fundamental issue about memory and its different forms is whether learning can occur without the development of conscious knowledge of what is learned. Amnesic patients and control subjects performed a serial reaction time task, exhibiting equivalent learning of an imbedded repeating sequence as measured by gradually improving reaction times. In contrast, four tests of declarative (explicit) knowledge indicated that the amnesic patients were unaware of their knowledge. Moreover, after taking the tests of declarative memory, all subjects continued to demonstrate tacit knowledge of the repeating sequence. This dissociation between declarative and nondeclarative knowledge indicates that the parallel brain systems supporting learning and memory differ in their capacity for affording awareness of what is learned. PMID- 10467600 TI - A stimulus paradigm inducing long-term desensitization of AMPA receptors evokes a specific increase in BDNF mRNA in cerebellar slices. AB - Long-term desensitization of AMPA receptors (LTDA) is a core mechanism of long term depression, a model of motor learning in the cerebellum. In this study we investigated the expression of neurotrophic factor genes after induction of LTDA in cultured cerebellar slices. LTDA was induced by application of quisqualate and monitored as a population response with a wedge recording technique. The levels of mRNA were quantified by reverse transcription followed by polymerase chain reaction. Quisqualate, at a dose and duration that reliably induced LTDA, elicited a significant and specific increase in BDNF mRNA with a peak at four hours after the application. By cell fractionation, the major source of BDNF mRNA increase was found to be in granule cells. In addition, a small but significant increase of transcripts with specific exon usage was observed in a Purkinje cell fraction. These results indicate that BDNF may be coinduced with LTDA and suggest that the slow and sustained increase of BDNF mRNA might play a role in later phases of synaptic plasticity in the cerebellum. PMID- 10467601 TI - Activity-dependent enhancement of presynaptic facilitation provides a cellular mechanism for the temporal specificity of classical conditioning in Aplysia. AB - A hallmark of many forms of classical conditioning is a precise temporal specificity: Learning is optimal when the conditioned stimulus (CS) slightly precedes the unconditioned stimulus (US), but the learning is degraded at longer or backward intervals, consistent with the notion that conditioning involves learning about predictive relationships in the environment. To further examine the cellular mechanisms contributing to the temporal specificity of classical conditioning of the siphon-withdrawal response in Aplysia, we paired action potential activity in siphon sensory neurons (the neural CS) with tail nerve shock (the US) at three critical time points. We found that CS-US pairings at short (0.5 sec) forward intervals produced greater synaptic facilitation at sensorimotor connections than did either 0.5-sec backward pairings or longer (5 sec) forward pairings, as reflected in a differential increase in both the amplitude and rate of rise of the synaptic potential. In the same preparations, forward pairings also differentially reduced the sensory neuron afterhyperpolarization relative to backward pairings, suggesting that changes in synaptic efficacy were accompanied by temporally specific changes in ionic currents in the sensory neurons. Additional experiments demonstrated that short forward pairings of sensory cell activity and restricted applications of the neuromodulatory transmitter serotonin (normally released by the US) differentially enhanced action potential broadening in siphon sensory neurons, relative to backward pairings. Taken together, these results suggest that temporally specific synaptic enhancement engages both spike-width-dependent and spike-width-independent facilitatory processes and that activity-dependent enhancement of presynaptic facilitation may contribute to both the CS-US sequence and proximity requirements of conditioning. PMID- 10467602 TI - Hitting the wall. PMID- 10467603 TI - Research funding opportunities at the Agency for Health Care Policy and Research. PMID- 10467604 TI - Ethics perspectives on end-of-life care. PMID- 10467605 TI - National survey of genetics content in basic nursing preparatory programs in the United States. PMID- 10467606 TI - Older Americans Act: implications for nursing. PMID- 10467607 TI - Active oxygen species generation and cellular damage by additives of parenteral preparations: selenium and sulfhydryl compounds. AB - We investigated the relationship between active oxygen species (AOS) generation and cultured vascular endothelial cellular damage caused by simultaneous exposure to selenium compounds and sulfhydryl compounds such as cysteine (Cys) or reduced glutathione (GSH). Selenium compounds, selenite, selenate or selenomethionine (SeMet), are added to total parenteral nutrition (TPN) and intravenously administered. We confirmed by luminol dependent chemiluminescence, an indicator of AOS generation, that selenite generates AOS in the presence of clinical concentrations of sulfhydryl compounds, 0.5 mM Cys or 0.5 mM GSH, and that the amount of AOS generated reaches the maximum when their mole ratio is 1:50. However, AOS generation was not observed after simultaneous administration of various concentrations of selenate or SeMet with sulfhydryl compounds. Moreover, simultaneous exposure to 10 microM selenite and sulfhydryl compounds was found to result in significant increases in the [3H]-adenine and lactate dehydrogenase (LDH) release rates from cells, a significant decrease in the amount of cellular protein, and enhancement of cellular damage as compared with after exposure to selenite alone. However, simultaneous exposure to 10 microM selenate or 10 microM SeMet together with sulfhydryl compounds did not induce cellular damage. These findings revealed that selenite generates AOS and causes cellular damage in the presence of sulfhydryl compounds. Accordingly, it seems better to choose selenate or SeMet instead of selenite when a selenium compound is to be added to TPN. PMID- 10467609 TI - Bone loss at the proximal femur and reduced lean mass following liver transplantation: a longitudinal study. AB - The longevity of recipients of liver transplant may be compromised by spinal osteoporosis and vertebral fractures. However, femoral neck fractures are associated with a higher morbidity and mortality than spine fractures. As there is little information on bone loss at this clinically important site of fracture, the aim of this study was to determine whether accelerated bone loss occurs at the proximal femur following transplantation. Bone mineral density and body composition were measured at the femoral neck, lumbar spine and total body, using dual x-ray absorptiometry in 22 men and 19 women, age 46 +/- 1.4 y (mean +/- SEM) before and at a mean of 19 mo after surgery (range 3-44). Results were expressed in absolute terms (g/cm2) and as a z score. Before transplantation, z scores for bone mineral density were reduced at the femoral neck (-0.47 +/- 0.21 SD), trochanter (-0.56 +/- 0.19 SD), Ward's triangle (-0.35 +/- 0.14 SD), lumbar spine (-0.76 +/- 0.13 SD), and total body (-0.78 +/- 0.15 SD) (all P < 0.01 to < 0.001). Following transplantation, bone mineral density decreased by 8.0 +/- 1.7% at the femoral neck (P < or = 0.01) and by 2.0 +/- 1.2% at the lumbar spine (P < or = 0.05). Total weight increased by 12.2 +/- 2.3%, lean mass decreased by 5.7 +/- 1.4%, while fat mass increased from 24.1 +/- 2.0% to 35.1 +/- 1.8% (all P < or = 0.001). Patients with end-stage liver disease have reduced bone mineral density. Liver transplantation is associated with a rapid decrease in bone mineral density at the proximal femur, further increasing fracture risk and a reduction in lean (muscle) mass, which may also predispose to falls. Prophylactic therapy to prevent further bone loss should be considered in patients after liver transplantation. PMID- 10467608 TI - Attenuation of the protein wasting associated with bed rest by branched-chain amino acids. AB - Bed rest is generally accepted as being an appropriate ground-based model for human spaceflight. The objectives of this study were to test the hypothesis that increasing the amount of branched-chain amino acids (BCAAs) in the diet could attenuate the protein loss associated with bed rest. Nineteen healthy subjects were randomized into two groups according to diet. During the 6 d of bed rest, the diets were supplemented with either 30 mmol/d each of three non-essential amino acids, glycine, serine, and alanine (control group), or with 30 mmol/d each of the BCAAs, leucine, isoleucine, and valine (BCAA group). Nutrition was supplied as a commercially available defined formula diet at a rate of 1.3 x REE. Nitrogen (N) balance and urinary 3-MeH excretion were determined for the 6 d. In our results, the urine-based estimate of N balance was 22.2 +/- 14.4 (n = 9) mg N.kg-1.d-1 and 60.5 +/- 10.1 mg (n = 8) N.kg-1.d-1 for the control and BCAA supplemented groups, respectively (P < 0.05). Urinary 3-MeH excretion was unchanged in both groups with bed rest. We conclude that BCAA supplementation attenuates the N loss during short-term bed rest. PMID- 10467610 TI - Use of TPN in terminally ill cancer patients. AB - Total parenteral nutrition (TPN) is often used as an adjunct to cancer therapy. However, it is increasingly being used in terminally ill cancer patients without clearly defined reasons. To determine the validity of the use of TPN in terminally ill cancer patients, 26 patients with limited life expectancy due to end-stage cancer were given TPN by their physicians, and the validity of its use was evaluated using the criteria of 1) quality of life, and 2) ultimate outcome. Patients were divided into two groups according to the use of TPN. Group I = TPN as adjunct of medical therapy, n = 15 (eight male, seven female), mean age 32 y. Group II = TPN for in-hospital supportive care, n = 11 (two male, nine female), mean age 56 y. Nutritional status on admission, quality of life (assessed by extent of daily activities, pain, and ability to sustain oral intake), and ultimate treatment outcome were determined. Mean weight loss in patients in Group I was 8.6 kg, 11 patients out of 15 were malnourished; mean weight loss in patients in Group II was 21 kg, and all of the 11 patients belonging to this group were malnourished. Two patients of Group I improved their quality of life, while 6 declined and 7 died; in Group II, 3 improved their quality of life, 4 declined, and 4 died. We conclude that when TPN was given either as an adjunct to in-hospital aggressive therapy for cancer or for in-hospital supportive care, quality of life did not improve in the majority of patients; nor did it influence ultimate outcome. These objective data, thus, raise the question of the validity of the use of TPN in terminally ill cancer patients. However, barring the cost factor, it is recognized that subjective reasons for giving TPN to terminally ill cancer patients persist and include compassionate, ethical, religious, or emotional reasons. PMID- 10467611 TI - Fat-free mass in chronic illness: comparison of bioelectrical impedance and dual energy x-ray absorptiometry in 480 chronically ill and healthy subjects. AB - Assessment of fat-free mass (FFM) and fat mass in ambulatory and hospitalized patients permits optimal adaptation of nutrition support. Recent methods for the determination of FFM are dual-energy x-ray absorptiometry (DXA) and bioelectrical impedance analysis (BIA). The purpose of this study was to determine if the BIA determined FFM as estimated by the formula by Kotler et al. and the Geneva formula could be validated when compared to DXA-determined FFM in subjects with various diagnoses. Body composition was measured by BIA and DXA in 480 subjects including healthy young men and women, elite female runners, and patients with various pathologies (including chronic obstructive and restrictive pulmonary disease patients; cystic fibrosis patients; lung, heart, and liver transplantation patients [both pre- and post-], and hemiplegic and AIDS patients). The present results suggest that BIA is relevant in the clinical assessment of body composition, but BIA formulas appropriate to the subject pool must be used for evaluation of FFM and fat mass. The BIA formula by Kotler et al. is appropriate for healthy subjects and patients with AIDS, cystic fibrosis, and pre- or postliver transplantation. The Geneva formula is more appropriate in pulmonary diseases and hemiplegic subjects with normal weight. PMID- 10467612 TI - Symptoms after total gastrectomy on food intake, body composition, bone metabolism, and quality of life in gastric cancer patients--is reconstruction with a reservoir worthwhile? AB - Gastric cancer is worldwide one of the most common causes of cancer death. Operation is the only treatment at this time that cures some patients. The side effects of the operation are, however, considerable, and include postoperative weight loss, loss of appetite, and other metabolic and nutritional changes. The recovery is very slow and incomplete. Reconstruction with different types of pouches has been asserted to facilitate the nutritional recovery, but results from different studies are somewhat contradictory. Malnutrition, osteoporosis, osteomalacia, and impaired quality of life are often but not always described. We can, however, establish that after a total gastrectomy, gastric cancer patients are very much at risk for these complications, which are probably caused by impaired food intake and steathorrhea even when the patients are cured from their cancer disease. In order to minimize the nutritional problems, it is crucial to avoid anastomotic narrowing and bile reflux. Roux-en-Y reconstruction seems to be the method of choice. Evidence from several randomized studies now speak in favor of including some type of pouch in the reconstruction. The most commonly used pouch today is the jejunal J-pouch. How the effect is exerted is not clear. Probably both the reservoir function of the pouch and changes in intestinal transit time are important. The importance of nutritional surveillance of these patients should not be underestimated, and most of the observed differences from various reports are probably due to dissimilarity in the follow-up protocols. A patient surviving his/her cancer has a decreased risk of developing severe disturbances in bone metabolism, food intake, body composition, and quality of life if the patient is under concerned nutritional surveillance and reconstructed with a pouch. PMID- 10467613 TI - Aluminum contamination of parenteral nutrition additives, amino acid solutions, and lipid emulsions. AB - Contamination of parenteral nutrition solutions with aluminum may result in accumulation of this element in bones and, in premature infants, may inhibit bone calcium uptake and induce cholestasis. We measured the aluminum concentration of small-volume parenterals, amino acid solutions, lipid emulsions, and special solutions containing glucose, amino acids, electrolytes, and trace elements (standard I for children with a body weight of 3-5 kg, standard II for children with a body weight of 5-10 kg). The method used was graphite furnace atomic absorption spectrometry GTA-AAS (SpectrAA-400 Plus, Varian, PtY Ltd., Mulgrave, Australia). Quality control was run with the use of control serum (Seronorm, Nycomed, Oslo, Norway). The aluminum contents of parenterally administered solutions were: pediatric trace elements, 130 micrograms/L, and pediatric trace elements, 3000 micrograms/L; phosphorus salts: K-phosphates, 9800 micrograms/L, and Na/K phosphates, 13,000 micrograms/L; 10% calcium gluconate, 4400 micrograms/L; 6.5% amino acids, 30 micrograms/L; 10% amino acids, 120 micrograms/L; 12.5% amino acids, 121 micrograms/L; 20% lipid emulsion, 30 micrograms/L; 20% lipid emulsion, 180 micrograms/L; water-soluble vitamins, 12 micrograms/L; lipid soluble vitamins, 360 micrograms/L; standard I, 55 micrograms/L; standard II, 90 micrograms/L; The aluminum intake from parenteral nutrition was 6.6-10.8 micrograms.kg-1.d-1--a dose exceeding the safety limit of 2 micrograms.kg-1.d-1. The possible association of aluminum not only with metabolic bone disease, but also with encephalopathy, dictates caution when dealing with the pediatric population on long-term parenteral nutrition. In the absence of reliable label information, it seems proper to monitor the aluminum concentration in parenteral nutrition products and to report it in professional journals. PMID- 10467615 TI - Soft tissue composition of pigs measured with dual x-ray absorptiometry: comparison with chemical analyses and effects of carcass thicknesses. AB - Evidence of the validity and accuracy of dual x-ray absorptiometry (DXA) to measure in vivo body composition is limited. We compared DXA estimates made in prone and side positions with measurements of chemical composition of 20 pigs (10 barrows and 10 gilts) weighing 52-113 kg. DXA yielded similar estimates of body composition in prone and side positions. DXA estimates of body composition were significantly correlated with reference compositional values (r2 = 0.927-0.998). No significant differences were found for determinations of body weight, fat mass (FM), fat free mass (FFM), bone-free, and fat-free mass (BFFFM) between DXA and chemical determinations. DXA significantly underpredicted percent fat (% fat); it underestimated FM (20%, P > 0.05), and overestimated FFM and BFFFM (6 and 9%, respectively, P > 0.05). Differences between individual determinations of FM and % fat by chemical analyses and DXA were significantly correlated with mean values. No significant correlations were found between the differences for weight, FM, % fat, FFM and BFFFM and measurements of carcass breadth (19-28 cm) and width (15-25 cm). Total errors in determination of DXA body composition variables were similar with body thicknesses less than and greater than 24 cm. These findings indicate that DXA is a valid and accurate method for determination of soft tissue composition. Initial problems with DXA determinations of % fat apparently have been reconciled partially with revisions in soft tissue analytic software. PMID- 10467614 TI - Effect of dietary glutamate on chemotherapy-induced immunosuppression. AB - Chemotherapy causes severe host immune depression and consequently increases susceptibility to infection. Dietary glutamate (GLU) serves as a stable substrate for the formation of glutamine (GLN), which is an important fuel and metabolic precursor for the immune cells. The effect of addition of GLU to a GLN/GLU-free amino acid diet upon immune response was studied in rats recovering from chemotherapy. Animals were fed a 0, 4, or 8% GLU diet and received a single intraperitoneal injection of methotrexate (MTX, 20 mg/kg BW). Two in vivo immune tests, delayed-type hypersensitivity (DTH) and popliteal lymphoproliferation (PLP), were performed 3 and 7 d after MTX treatment. Food intake and body weight decreased significantly immediately after MTX treatment and gradually recovered after 8 d with no significant difference among treatment groups. In a 23-d feeding study, no significant difference was found in the DTH response, but the PLP response increased in a GLU dose related fashion (83 and 133% increases for the 4 and 8% GLU diets, respectively). In a 44-d feeding study, the DTH response increased 61 and 83%, while the PLP response increased 191 and 382% for the 4 and 8% GLU diets, respectively. Plasma GLN, GLU, or glutathione (GSH) levels were increased by dietary GLU, but only in the immediate postprandial state. In summary, dietary GLU improves immune status of rats recovering from MTX treatment. The immune-enhancing effect of dietary GLU was dose-dependent and more pronounced after a longer duration of dietary GLU intake. PMID- 10467616 TI - Control of food intake by fatty acid oxidation and ketogenesis. AB - Fatty acid oxidation seems to provide an important stimulus for metabolic control of food intake, because various inhibitors of fatty acid oxidation (mercaptoacetate, methyl palmoxirate, R-3-amino-4-trimethylaminobutyric acid) stimulated feeding in rats and/or mice, in particular when fed a fat-enriched diet, and long-term intravascular infusion of lipids reduced voluntary food intake in various species, including humans. The feeding response to decreased fatty acid oxidation was due to a shortening of the intermeal interval with meal size remaining unaffected. Thus, energy derived from fatty acid oxidation seems to contribute to control of the duration of postmeal satiety and meal onset. Since inhibition of glucose metabolism by 2-deoxy-D-glucose affects feeding pattern similarly, and spontaneous meals were shown to be preceded by a transient decline in blood glucose in rats and humans, a decrease in energy availability from glucose and fatty acid oxidation seems to be instrumental in eliciting eating. Since the feeding response of rats to inhibition of fatty acid oxidation was abolished by total abdominal vagotomy and pretreatment with capsaicin destroying non-myelinated afferents and attenuated by hepatic branch vagotomy, fatty acid oxidation in abdominal tissues, especially in the liver, apparently is signalled to the brain by vagal afferents to affect eating. Brain lesions and Fos immunohistochemistry were employed to identify pathways within the brain mediating eating in response to decreased fatty acid oxidation. According to these studies, the nucleus tractus solitarii (NTS) of the medulla oblongata represents the gate for central processing of vagally mediated afferent information related to fatty acid oxidation. The lateral parabrachial nucleus of the pons seems to be a major relay for pertinent ascending input from the NTS. In particular the central nucleus of the amygdala, a projection area of the parabrachial nucleus, appears to be crucial for eating in response to decreased fatty acid oxidation. As ketones are products of hepatic fatty acid oxidation that are released into the circulation and peripheral (and central) administration of 3-hydroxybutyrate reduced voluntary food intake in rats, ketones being utilized as fuels by the peripheral and central nervous system might contribute to control of eating by fatty acid oxidation, especially when high levels of circulating ketones occur. Whether a modulation of the hepatic membrane potential resulting from changes in the rate of fatty acid oxidation and/or ketogenesis represent a signal for control of eating transmitted to the brain by vagal afferents remains to be established. Recent in vivo studies investigating the effects of mercaptoacetate on the hepatic membrane potential and on afferent activity of the hepatic vagus branch are consistent with this notion. Further investigations are necessary to delineate the coding mechanisms by which fatty acid oxidation and/or ketogenesis modulate vagal afferent activity. PMID- 10467617 TI - Aluminium contamination of parenteral fluids, need for product label information. PMID- 10467618 TI - Phagocyte-induced lipid peroxidation of intravenous fat emulsions: is there a need for reformulation? PMID- 10467619 TI - Nucleotides and insulin secretion. PMID- 10467620 TI - Selenium and HIV infection. PMID- 10467621 TI - Chromium and diabetes. PMID- 10467622 TI - A complication of long-term brain microdialysis. PMID- 10467623 TI - The not-so-missing link between nutrition support nurses and case managers. PMID- 10467624 TI - Nutritional aspects of liver disease in children. PMID- 10467625 TI - Drug selection and the management of corticosteroid-related diabetes mellitus. AB - Glucocorticoid use is associated with the risk of hyperglycemia in patients without known diabetes mellitus and worsened glycemic control in diabetic patients. The effects are greater in the fed than fasting state. Management includes use of diet and exercise (as appropriate for the individual) in all patients. Mild hyperglycemia can often be managed with oral agents, especially those with rapid onset of action. Marked hyperglycemia, especially in diabetic patients or patients with liver or renal disease, requires insulin. Adjustments in insulin can be done both in anticipation of the glucocorticoid effect and based on home glucose monitoring. The effects of glucocorticoids on hyperglycemia usually remit within 48 hours of discontinuation of oral administration. PMID- 10467626 TI - Management of lipid disorders. AB - ASCVD is common in patients with rheumatologic disorders. Reduction of LDL cholesterol and treatment of lipid disorders is proved to reduce the risk of ASCVD and its associated clinical events. Therefore, plasma lipids should be obtained in all patients with rheumatologic disorders and lipid disorders should be aggressively treated in an attempt to reduce cardiovascular risk. The clinical approach is similar to other patients, but care should be taken to avoid side effects and drug-drug interactions, which may be somewhat more likely to occur in patients with rheumatologic disorders. PMID- 10467627 TI - Evaluation and treatment of hypertension. AB - Hypertension is a significant and prevalent risk factor for the development of cardiovascular disease and target organ damage. The urgency of treatment of high blood pressure depends on the level of blood pressure elevation and the presence of coexistent risk factors for cardiovascular disease. Likewise, the level to which blood pressure is reduced is not restricted to the definition of high blood pressure but instead depends on the underlying disease. Diabetes and renal insufficiency, for example, require blood pressure goals below those that are traditionally defined. In the absence of contraindications, beta-blockers and diuretics are still recommended as first-line agents for treatment of uncomplicated hypertension. Calcium channel antagonists also may reduce mortality. In patients with diabetes, ACE inhibitors are effective first-line agents in type 1 and type 2 diabetic patients who are hypertensive or have microalbuminuria. ACE inhibitors may be beneficial in patients with nondiabetic renal insufficiency as well. Calcium channel antagonists may have some effect in retarding progression of diabetic nephropathy although a recent trial found a higher incidence of death as a secondary endpoint in hypertensive diabetic patients who were treated with calcium channel antagonists. Beta-blockers seem to be safe and well tolerated in patients with mild to moderate intermittent claudication, although patients with rest pain or limb ischemia have not been studied. Beta-blockers should not be used in patients with asthma. Dihydropyridine calcium channel antagonists are the preferred treatment of hypertension in patients with Raynaud's but should be avoided in patients with severe gastroesophageal reflux disease. NSAIDs, particularly piroxicam and indomethacin, raise mean blood pressure by approximately 5 mm Hg, enough to consider a change of either NSAID or antihypertensive to one that is not as affected by NSAIDs. Cyclosporine A can induce hypertension by its vasoconstrictive effects, particularly on the kidney. Calcium channel antagonists may antagonize this vasoconstriction while allowing the clinician to reduce the dose of cyclosporine A required to achieve its immunosuppressive effect. PMID- 10467628 TI - Women's health. AB - Symptoms reliably attributable to menopause are vasomotor symptoms and vaginal dryness. Other symptoms are not directly related to the menopause. HRT or ERT are effective in providing symptom relief and preventing disease prevalent in postmenopausal women. HRT or ERT is beneficial in women with RA. Little data concerns safety in SLE patients, but there are theoretical advantages to using HRT or ERT in women with SLE and two studies that indicate it is safe. Other forms of treatment are available to women who cannot or will not use HRT or ERT. PMID- 10467629 TI - Vaccination of the immunosuppressed adult patient with rheumatologic disease. AB - Vaccine-preventable diseases are a major cause of morbidity in immunocompromised patients. This article reviews the data on efficacy and safety of currently licensed vaccines in patients with rheumatologic diseases. PMID- 10467630 TI - Recognition and management of preoperative risk. AB - Internists are frequently asked to do preoperative consultations and to manage perioperative complications. Realistic goals are to identify patient factors that increase the risk of surgery, to quantify this risk in order to make decisions about the appropriateness of and timing of the surgery, to provide recommendations on how to minimize the risk, to identify and manage coexisting medical conditions and their associated medication requirements, to monitor the patient for perioperative problems, and to make recommendations to deal with these problems when they occur. With few exceptions, nonselective imaging and laboratory screening tests have repeatedly been shown to be of little value when the history and physical do not suggest a problem. The risk associated with the planned surgery can be estimated, with the most common serious complications being cardiac events. Updated versions of Goldman's risk indices are particularly helpful for this. Clinical variables are optimally combined with selective stress testing to discern which patients will benefit from preoperative revascularization. This has been studied best in the setting of vascular surgery. A critical guiding principle is that the value of revascularization must be judged in terms of long term gains rather than just immediate perioperative benefit. Other interventions include the selective use of beta blockers, adequate analgesia for all, control of hypertension, and appropriate volume management, especially in the settings of preexisting CHF or valvular disease. It must also be recognized that perioperative ischemia and CHF often present atypically. An approach that combines aspects of both the ACC/AHA and the ACP guidelines seems optimal. A variety of noncardiac issues must also be addressed. Postoperative pulmonary complications are common, especially with preexisting pulmonary disease, thoracic and upper abdominal surgery, and obesity. PFTs and ABGs are indicated in selected patients. Stopping smoking, incentive spirometry, and selective use of bronchodilators and antibiotics are helpful. Patients with rheumatologic diseases have specific concerns based on systemic manifestations of disease including anemia, thrombocytopenia, pulmonary fibrosis, pericarditis, and hypercoagulability; medication effects particularly from steroids and nonsteroidal anti-inflammatory drugs; and specific joint problems including contractures and atlantoaxial joint instability. Diabetes increases the risk of infection and cardiac complications. Prevention of ketoacidosis and glucose control are necessary and can be achieved through a variety of approaches, depending on whether the patient suffers from Type 1 or Type 2 diabetes. The threshold for transfusion has increased in recent years, as has the use of erythropoietin and autologous blood donation. There is no longer an absolute hemoglobin that requires transfusion, although most require transfusion for hemoglobins less than 8 mg/dL, especially in the setting of cardiac disease and bloody surgery. The elderly require surgery at an increased rate and often do not do as well as younger patients. The primary issues are, however, not their age but their increased frequency of underlying disease and diminished reserve. The latter makes them prone to postoperative delirium, sensitivity to medications, and cardiac and pulmonary problems. Despite the many diseases that patients often have and the stresses of surgery itself, modern anesthetic and surgical techniques allow almost all patients to undergo necessary procedures at acceptable risk. The internist plays a critical role in minimizing this risk even further. PMID- 10467631 TI - Perioperative management of selected problems in patients with rheumatic diseases. AB - Patients undergoing surgery are subject to multiple perioperative problems. This article reviews several issues that occur in surgical patients with rheumatic diseases, including management of medications, diagnosis of fat embolism syndrome, prophylaxis against endocarditis, postoperative fever, and perioperative myocardial infarction. PMID- 10467632 TI - Deep vein thrombosis and pulmonary embolism prophylaxis in joint replacement surgery. AB - Joint replacement surgery is one of the most frequently performed procedures in the United States. The incidence of deep vein thrombosis and pulmonary embolism is very high in patients not receiving prophylaxis for the prevention of this postoperative complication. In this article, the current modalities for prophylaxis are reviewed with respect to their safety and efficacy. Recommendations that have been substantiated by evidence-based information are provided. PMID- 10467633 TI - Detection and management of coronary artery disease in patients with rheumatologic disorders. AB - Coronary heart disease causes significant morbidity and mortality in our society and is more common in patients with some rheumatic disorders. A thorough history is the most essential element in assessing patients with chest pain syndrome or dyspnea who are at risk of harboring coronary artery disease. Based on the level of suspicion, a noninvasive test is commonly ordered to confirm the diagnosis. If a patient is active and can exercise, then treadmill or bike stress echocardiography is recommended. In addition to assessment of valvular, pericardial, and myocardial function at rest, stress echocardiography permits direct visualization of left ventricular cavity enlargement or wall motion abnormalities, which imply significant myocardial ischemia. If the echocardiography laboratory does not have sufficient expertise or experience with this technique, then exercise thallium is the next study of choice. If a patient is inactive or is unable to exercise, then dobutamine stress echocardiography is recommended. For patients who are in atrial fibrillation, have permanent pacemakers, who cannot augment their heart rate to a minimum of 85% MPHR, dipyridamole thallium is an acceptable alternative. An ischemic response in a symptomatic patient in most cases requires further evaluation with cardiac catheterization. Based on the patient's clinical presentation and physiologic and anatomic findings, important therapeutic decisions can then be made. PMID- 10467634 TI - Large artery occlusive disease. AB - Large artery occlusive disease is a common problem in the United States. It affects both the upper and lower extremities and is associated with significant morbidity and mortality. This article deals with the clinical recognition of this entity in hopes that the general internist and rheumatologist will more easily recognize it. In addition, the latest technology available to diagnose and treat large artery occlusive disease is discussed. PMID- 10467635 TI - Depression in the patient with rheumatologic disease. AB - Depression is commonly seen in patients with chronic disease, including rheumatologic disease. Because of ongoing contact with their patients, rheumatologists are uniquely poised to recognize depressive disorders and formulate a treatment plan. Recent advances in pharmacotherapy have simplified management and increased the likelihood of successful treatment. Through heightened awareness, rheumatologists can feel more comfortable in diagnosing and initiating therapy for depression. PMID- 10467636 TI - Dyspepsia and heartburn. AB - Dyspepsia and heartburn are the two cardinal symptoms of foregut dysfunction. When confronting such a problem, that physician must first learn to discern between the two, because treatment can be quite different for the conditions presenting with these symptoms. This article details the approach to work-up and treatment of patients presenting with dyspepsia or heartburn. PMID- 10467637 TI - Screening for cancer in the patient with rheumatic disease. AB - This article begins with a discussion of the types of cancer that are most often the target of cancer screening in general medical practice. Colorectal, prostate, cervical, and breast cancer screening guidelines are reviewed and some of the evidence (and controversy) behind these recommendations is discussed. A discussion of some of the special risks for cancer in patients with rheumatologic disease follows, and some modifications of standard screening guidelines are suggested for these high-risk patients. PMID- 10467638 TI - Selected drug complications and treatment conflicts in the presence of coexistent diseases. AB - The presence of different coexistent systemic diseases often times complicates the selection of the appropriate treatment of an underlying rheumatologic condition. In this article, some controversial treatment conflicts that are frequently encountered in the daily practice of rheumatology are clarified and guidelines for the best available therapeutic options are provided. PMID- 10467639 TI - Bacterial resistance, beta-lactam antibiotics and gramnegative bacteria at ICUs. PMID- 10467640 TI - [Intestinal ischemic reperfusion syndrome: pathophysiology, clinical significance, therapy]. AB - Gut ischemia-reperfusion injury is a serious condition in intensive care patients. Activation of immune cells within the huge endothelial surface area of gut microcirculation may initiate a systemic inflammatory response with secondary injury to distant organs. Translocation of bacteria and toxins through a leaky gut mucosa may amplify or perpetuate systemic inflammation, leading to multiple organ dysfunction syndrome and death in critically ill patients. Gut ischemia promotes regional production of inflammatory mediators, expression of cell adhesion molecules on endothelial and immune cell surfaces and increases the procoagulatory properties of vascular endothelial cells. During reperfusion, gut injury may be amplified by increased production of oxygen radicals and exhaustion of endogenous antioxidant defence mechanisms. Although several therapeutic strategies to interrupt the pathophysiology of ischemia-reperfusion have been shown to be beneficial in animal experiments, none of these interventions has gained clinical relevance. After initial hemodynamic and respiratory stabilisation of critically ill patients, strategies to prevent secondary gut injury by increasing splanchnic oxygen delivery or augment mucosal cell regeneration may be the only therapeutic options for intensive medical specialists at the present time. Early enteral nutrition and treatment with specific vasoactive drugs may reduce morbidity and costs of treatment in certain critically ill patients. However definitive evidence of a reduction in mortality with these therapies has still not be provided. PMID- 10467641 TI - In vitro activity of newer broad spectrum beta-lactam antibiotics against enterobacteriaceae and non-fermenters: a report from Austrian intensive care units. Austrian Carbapenem Susceptibility Surveillance Group. AB - We compared the in vitro activity of broad spectrum beta-lactam antibiotics against 573 gram-negative isolates (enterobacteriaceae and non-fermenters) collected between November 1996 and May 1997 from 9 laboratories serving intensive care units throughout Austria. MIC's (Minimal Inhibitory Concentration) were obtained with the E-test for meropenem, imipenem, ceftazidime, cefepime, cefpirome and piperacillin/tazobactam. Pseudomonas aeruginosa was the most frequently isolated organism (22%), followed by E. coli (19%), Klebsiella spp. (16%), and Enterobacter spp. (14%). Acinetobacter spp., Proteus spp., Serratia spp., Stenotrophomonas maltophilia, Citrobacter spp., Morganella morganii, Burkholderia cepacia and Salmonella enteritidis were isolated less frequently. Overall meropenem, imipenem and ceftazidime were the most active compounds in vitro, inhibiting 90%, 89%, and 87% of the isolates, respectively. Pseudomonas aeruginosa was inhibited by piperacillin/tazobactam in 89%, by cefepime in 87% and by ceftazidime in 85%. Imipenem, meropenem and cefpirome were less active (79%, 75% and 69% respectively). All E. coli strains were inhibited by meropenem, 99% were inhibited by imipenem, cefepime and cefpirome. Ceftazidime was active against 95% and piperacillin/tazobactam against 92% of E. coli. All Klebsiella spp. were inhibited by meropenem, cefepime and cefpirome. Imipenem inhibited 99% and ceftazidime 98% of the Klebsiella isolates. Piperacillin/tazobactam was active against 95% of Klebsiella spp. In vitro carbapenems are still the most active of all antibiotics tested. The relatively high resistance of Pseudomonas spp. and Acinetobacter spp. to carbapenems reflects the wide use of carbapenems during the last years. However, most bacterial isolates are still sensitive to the tested broad spectrum beta-lactams. PMID- 10467642 TI - [Difference between gastric mucosal pCO2 and arterio-intramucosal pCO2 during orthotopic liver transplantation]. AB - Tonometry is a clinically accepted method to monitor blood flow of the splanchnic region, which is of particular interest in orthotopic liver transplantation (OLT). We investigated the hemodynamic changes and the tonometrically registered perioperative course of the difference between gastric mucosal pCO2 (prCO2) and arterial mucosal CO2 (CO2 gap) in 23 patients undergoing OLT without veno-venous bypass. Gastric mucosal pH (pHi) was additionally calculated. Despite significant changes in systemic hemodynamics during the anhepatic stage and after reperfusion and a significant drop in pHi during anhepacy, the difference between prCO2 and CO2 was constant. These contrasting findings of tonometry, i.e. solely a drop in pHi is, in our opinion, a consequence of the poor metabolic capacity of the liver in the perioperative OLT period, which influenced the calculation of the pHi with the Henderson-Hasselbalch equation. We conclude that, due to methodical problems, calculated pHi is not a reliable indicator of splanchnic blood flow and oxygenation during OLT. We therefore suggest that the prCO2 and the CO2 gap be used to monitor the splanchnic region. These parameters, obtained perioperatively, do not indicate a further reduction in splanchnic oxygenation despite profound changes in systemic hemodynamics during OLT without veno-venous bypass. PMID- 10467643 TI - [Interdisciplinary surgical treatment of anterior skull base tumors]. AB - OBJECTIVES: We present our clinical experiences regarding interdisciplinary surgical treatment of anterior skull base tumours and evaluate postoperative results. METHODS: Fifty-seven patients (25 male, 32 female) with benign and malignant neoplasms involving the anterior skull base were retrospectively reviewed. In all cases tumour resection was carried out by an interdisciplinary rhino-neuro-surgical skull base operating team. Forty-three of 57 patients (75.4%) underwent common transbasal tumour resection and 11 (19.3%) were operated on from an extended transbasal approach. An extensive transbasal approach for tumour resection was used in 3 patients (5.3%). Postoperative mortality and morbidity were evaluated over a period of 6 months. RESULTS: In all 57 patients a good access to the frontal fossa and the sinuses was achieved. By means of the transbasal approaches, one-step tumour removal was possible in all cases. Tumor diameter ranged fom 12 mm to 114 mm. Even tumours extending as far as the hard palate required no additional transfacial procedures. Surgical mortality was 3.5%. Permanent postoperative complications were noted in 4 cases (7.02%) and transient postoperative complications in 7 (12.28%). CONCLUSION: In dealing with anterior skull base tumours, interdisciplinary surgical procedures using transbasal approaches provide a satisfactory outcome at a low rate of postoperative complications. When transbasal approaches are applied, no additional transfacial skull base exposure using midfacial incisions is required. PMID- 10467644 TI - [Diagnosis and therapy of common bile duct stones in a patient with a history of hypersensitivity reaction to radiologic contrast media --a case report]. AB - Common bile duct (CBD) stones may occur in patients who had been previously cholecystectomized because of gallbladder stones. CBD stones occur in 15%-20% of patients with symptoms secondary to cholecystolithiasis. After cholecystectomy they occur in 1%-5% of patients, either in the form of retained or recurrent choledocholithiasis. Endoscopic ultrasonography (EUS) is a novel and excellent diagnostic screening procedure for common bile duct stones. Numerous reports confirm an equal success rate for endoscopic ultrasound and endoscopic retrograde cholangiopancreatography (ERCP). However, unanimity of opinion with respect to the best diagnostic procedure for the detection of CBD stones does not exist to date. We report a 74-year-old female patient with CBD stones, who was shown to have a severe allergy to X-ray contrast medium at a previous examination. In view of the pre-existing contraindication for ERCP, we diagnosed the stones by EUS. The majority of patients would require ERCP for endoscopic treatment, but our patient rejected the administration of contrast medium as well as conventional surgical treatment. After introducing a guide wire into the bile duct to assure its position, endoscopic sphincterotomy (EST) without contrast imaging was performed successfully. The CBD stones were extracted with a basket. Based on our results we believe that EUS ranks as an important procedure in the diagnosis of CBD stones, and that successful endoscopic treatment can be performed without previous ERCP. We do realize that EST without ERCP in unsuitable as a routine procedure, but may be used successfully in specific cases such as that of the patient reported here. PMID- 10467645 TI - [Brugada-Brugada syndrome. An atypical case]. AB - The Brugada-Brugada syndrome is a rhythmologic disorder which can be diagnosed because of typical ECG criteria. A high-take off descending ST segment localized to the right chest leads, associated with right bundle branch block and ventricular fibrillation or syncopes are characteristic of the syndrome. ECG alterations in the right precordial leads were recorded in a 47 year old female patient who was admitted to hospital because of enteritis and associated syncope. The ECG alterations were initially not realized as Brugada-Brugada syndrome. Because of "recognizing" comparable ECG alterations during a congress lecture, the diagnosis was made. The patient was treated with an ICD. PMID- 10467646 TI - [Cardiovascular and metabolic response to dynamic stress echocardiography by patients with coronary heart disease and healthy probands]. AB - 25 patients with coronary heart disease (CHD, 62 +/- 9 years) and 18 subjects free from cardiovascular disease (28 +/- 9 years) were tested on a cycle ergometer using a graded incremental test protocol: a) in the common upright position, b) as dynamic stress echocardiography in a semi-supine position. Whereas no relevant differences could be detected between the two conditions for the healthy subjects concerning heart rate, blood pressure, and rate-pressure product (2 x 3 ANOVA, t-tests with alpha-adjustment), the CHD patients showed both significantly higher heart rates as well as a significantly higher rate pressure product (50 W: 15,300 +/- 2973 mm Hg/min vs. 13,822 +/- 3042 mm Hg/min; 75 W: 18,028 +/- 3479 mm Hg/min vs. 16,337 +/- 2619 mm Hg/min) on equivalent stages during stress echocardiography if compared to the sitting position. There were no differences for systolic blood pressure in this group; the diastolic values were higher in the sitting position at rest and during 50 W. Lactate concentrations (determined only in the healthy subjects) were significantly higher on all stages during dynamic stress echocardiography. The workload at the individual anaerobic threshold (IAT) was significantly lower. In conclusion, we found a higher metabolic along with a similar cardiovascular strain at equivalent workloads in stress echocardiography compared to upright bicycle ergometry for healthy subjects. However, CHD patients have a higher cardiocirculatory load in the semi-supine position. When investigating these patients with stress echocardiography, higher heart rates of about 8 beats/min have to be expected for equivalent workloads if compared to the upright position. PMID- 10467647 TI - [Hybrid technique for myocardial revascularization: value of combined minimal invasive bypass technique (MIDCAB) with interventional therapy (PTCA)]. AB - BACKGROUND: Minimally invasive direct coronary artery bypass (MIDCAB) grafting without cardiopulmonary bypass (CPB) through an anterolateral minithoracotomy has become a promising therapeutical option especially in multimorbid, elderly and reoperative patients with single vessel disease. However, this procedure precludes complete revascularization in multivessel disease because the minithoracotomy limits the surgical access either to anterior or lateral or posterior vessels of the beating heart. To expand the benefits of the MIDCAB concept to patients with multivessel disease, new interdisciplinary approaches have recently been introduced. METHODS: Since December 1996, 26 patients (21 male, 5 female, mean age 56.6 +/- 18.8 years) underwent a "hybrid" revascularization performed as a primary MIDCAB procedure for grafting of the left anterior descending artery (LAD) with the left internal mammary artery (LIMA) followed by staged angioplasty and stenting of additional coronary lesions. RESULTS: After MIDCAB grafting, the postoperative course was uneventful in all patients. Coronary re-angiography after a median of 7 days revealed patent and functioning LIMA grafts in all patients. Applying subsequent percutaneous transluminal coronary angioplasty (PTCA) and occasional stenting (n = 8), a total of 31 lesions were treated successfully. Procedure related complications did not occur. All patients remained angina-free and no stress electrocardiographic changes were recorded. CONCLUSION: Our preliminary results of a "hybrid" approach to myocardial revascularization suggest that this concept is a safe and effective approach of complete revascularization for selected patients with multivessel involvement. Especially elderly and reoperative patients with significant comorbidity may benefit from hybrid procedures avoiding cardiopulmonary bypass and midsternotomy. PMID- 10467648 TI - [Long-term outcome of therapy of recurrent myocardial ischemia after surgical revascularization]. AB - Although there are randomized data for CABG vs. medical therapy and CABG vs. PTCA in primary therapy of CAD, there is few evidence on the appropriate therapy of recurrent angina after prior CABG. We analyzed data from 1265 consecutive patients (pts.) presenting for recurrent angina after previous CABG who required either re-CABG (n = 67), PTCA (n = 768) or medical therapy alone (medRx) (n = 430) at our institution during 1986 through 1996. Outcome after index therapy is monitored during 45 +/- 34 months. The 3 therapeutic groups were similar with respect to gender (84% male), age at therapy (60 years), prevalence of diabetes (22%), and time to first ischemic event after primary CABG (37 months). Actuarial survival was significantly higher in the PTCA-group at 1, 5 and 10 years after therapy of recurrent angina, despite the freedom from subsequent re-intervention was significantly lower (1-year-survival 95% [37%] vs. 95% [3%] medRx vs. 79% [4%] re-CABG, 5-year-survival 87% [57%] vs. 78% [17%] medRx vs. 50% [4%] re-CABG and 10-year-survival 72% [65%] vs. 63% [31%] medRx vs. 50% [15%] re-CABG resp., p < 0.0001, [numbers in brackets represent corresponding values for incidence of re intervention, p < 0.0001]). These findings were similar after adjustment for different baseline characteristics. Following this adjustment multivariate Cox analysis identified age beyond 70 years, diabetes mellitus and therapy: redo-CABG as independent correlates for mortality for the entire group. THERAPY: angioplasty was identified as an independent correlate for survival. In contrast, therapy: angioplasty was an independent correlate for re-intervention after therapy. CONCLUSIONS: In this nonrandomized series of patients with recurrent angina after previous CABG, an initial strategy of angioplasty resulted in a significant higher overall survival, although this regimen is associated with a greater need for subsequent revascularization procedures. PMID- 10467649 TI - [Coronary vasculopathy after heart transplantation--effect of temporal onset, severity and progression on long-term prognosis]. AB - Despite considerable progress in the knowledge about pathophysiology of cardiac allograft vascular disease (CAVD), only few systematic studies are available, characterizing the natural course during long-term follow-up after heart transplantation (HTX). Therefore, we analyzed in 354 heart transplant recipients (305 male, 45.9 +/- 11.2 years, mean observation period 5.8 +/- 3.0 years, range 0.9-12.4 years) the results of 1129 coronary angiograms under the aspects of development, severity, localization, and progression of disease related to the prognosis of patients. As expected an increasing prevalence was found over time with a luminal obstruction > or = 30% in 83% of all patients more than 10 years after HTX. Coronary artery stenosis (> or = 50%) at initial diagnosis was predominantly localized in the LAD (46%) followed by RCX (31%) and RCA (23%). Angiographic risk profiles with an impaired prognosis could be identified in the form of an early development (< 4 years post HTX) of disease (p = 0.03), luminal obstruction > 50% (p = 0.001), and multivessel appearance at first diagnosis (p = 0.02) as well as in progressive forms of CAVD (p = 0.001). Summarizing, CAVD is a frequent finding in HTX recipients. Especially in patients with early onset, progressive, and advanced stages of disease it represents a prognostically limiting complication following HTX. Identification of the natural course is of major importance defining the need and efficacy of future palliative therapeutical approaches. PMID- 10467650 TI - [Quantitative detection of changes in the thoracic aorta in patients with chronic aortic dissection using transesophageal echocardiography]. AB - The aim of this serial 3 year follow-up study in 42 clinically stable patients with chronic aortic dissection was to assess quantitatively morphologic changes of the descending thoracic aorta (AD) using transesophageal echocardiography (TEE). Communicating dissections (ca) were present in 16/19 patients with operated type I and in 11/23 patients with type III AD whereas 12/23 type III AD according to De Bakey were non-communicating (nc). Diametral enlargement of the disc. thoracic aorta was 4 mm (mean value) at 1 year in all patients, 5.9 mm in type I ca, 7.2 mm in type III ca but only 3.1 mm in type III nc at 3 years. The ratio between true lumen and false lumen (FL) changed in ca AD from 1:2 to 1:3 over the period of 3 years but remained constant at 1:1 in ncAD. Progressive thrombosis of the false lumen (FL) occurred in 76% of patients but complete thrombosis of the FL occurred in only 6% of type I ca, 18% type III ca but in 84% of type III nc patients. Our results confirm observations that non-communicating dissections seem to have a more favorable outcome and less aneurysmal dilatation compared to ca dissection. PMID- 10467651 TI - [Interventional occlusion of persistent ductus arteriosus Botalli with Gianturco spirals]. AB - Interventional duct occlusion is increasingly performed with detachable coils. In a series of 33 patients (34 procedures), we consistently used Gianturco-coils which were the initially employed coils. They are not securable but can be implanted in a simple manner. The rate of embolization was high (28%) in the first 14 procedures; however, it declined to 5% in the last 20 patients. We modified the technique by partially blocking the duct, when necessary, with a catheter which was advanced from the pulmonary artery and by implanting up to three coils. There was only one embolization and one small residual shunt in the last 20 patients. The mean fluoroscopy time in these patients was 11.0 +/- 5.8 minutes and decreased to 7.0 +/- 3.2 minutes in the last 10 patients with successful closure. Considering our experience, we continue to recommend the use of Gianturco-coils especially in the small duct which can be closed quickly at low cost. PMID- 10467652 TI - [Implantation of a dual chamber pacemaker-defibrillator (DDD-ICD) in a patient with hypertrophic obstructive cardiomyopathy]. AB - A 70-year-old woman with severely symptomatic hypertrophic obstructive cardiomyopathy was unresponsive to drug treatment. She had recurrent ventricular tachyarrhythmias and syncope and was at high risk for sudden death; a dual chamber pacemaker defibrillator (DDD-ICD) was implanted. Her initial left ventricular outflow tract gradient was 80 mm Hg and fell to 40 mm Hg during dual chamber pacing at an atrial ventricular delay of 140 ms. In the follow-up over six months she was asymptomatic with respect to angina pectoris; ventricular tachycardias could be successfully terminated by antitachycardia pacing or by shocks. A dual chamber pacemaker defibrillator is an important therapeutic option for patients with symptomatic hypertrophic obstructive cardiomyopathy and ventricular tachyarrhythmias. PMID- 10467653 TI - [Catheter-induced dissection of the right coronary ostium and aortic root]. AB - We describe a catheter-induced dissection of the ostium of the right coronary artery extending to the aortic root. As an alternative to emergency surgery, a dacron stent-graft was placed in the proximal right coronary artery, thus, sealing the dissection. The patient later underwent elective aortocoronary bypass surgery. The therapeutic options in this situation with diffuse coronary disease, in which only one severe culprit lesion can be identified, are discussed. PMID- 10467655 TI - [Current status of education and continuing education of nonmedical assistant personnel in interventional cardiology]. PMID- 10467654 TI - [Follow-up of dissection of the right coronary artery and wall of the ascending aorta after directional coronary atherectomy. A case report]. AB - We report on a patient in whom a restenosis after two PTCA procedures was treated with directional coronary atherectomy. In the second DCA maneuvre a spiral dissection, type D, developed from ostium to restenosis with a balloon shaped part in the wall of the aorta ascendens. This part disappeared nearly completely by next day. The dissection in the right coronary artery persisted three months later until a bypass operation and was not visible after two years. PMID- 10467656 TI - [In Process Citation] PMID- 10467657 TI - Ferdinand Cohn 1828-1898. In commemoration of the 100th anniversary of his death. PMID- 10467658 TI - Characterization of nosocomial Serratia marcescens isolates: comparison of Fourier-transform infrared spectroscopy with pulsed-field gel electrophoresis of genomic DNA fragments and multilocus enzyme electrophoresis. AB - A total of 66 Serratia marcescens isolates from 46 patients was investigated by macrorestriction using XbaI followed by pulsed-field gel electrophoresis. 7 restriction fragment patterns attributable to more than one patient and 9 individual patterns were identified. The isolates were additionally characterized by multilocus enzyme electrophoresis and Fourier-transform infrared spectroscopy. The macrorestriction patterns and the multilocus enzyme electrophoresis patterns corresponded fairly well while the classifications derived from these methods were not completely congruent. The grouping achieved by Fourier-transform infrared spectroscopy on the basis of high (> 1000) and moderately high heterogeneity values (300) was consistent with the macrorestriction results. Grouping on a lower heterogeneity level did not contribute to further discrimination. In general, Fourier-transform infrared spectroscopy was less discriminatory than the two other methods, but easier to perform. Therefore, laboratories equipped with the necessary devices may use it to rapidly select bacterial isolates for macrorestriction or other well established characterization procedures. PMID- 10467659 TI - Taxonomic study and morphological differentiation of an actinomycete genus, Kitasatospora. AB - The history and taxonomic criteria of the actinomycete genus, Kitasatospora, are described. The reasons for a change of the position of the genus in the classification system are given and a restriction length polymorphism (RFLP) method for rapid identification is described. Although the bacterial genus, Kitasatospora, resembles the fungal one, Streptomyces, in its morphology, it is clearly different in its cell-wall composition which includes LL- and meso-DAP, glycine and galactose. Following a change in its taxonomic position as a result of insufficient gene analysis, it was re-established by phylogenetic analysis of the entire 16S rRNA gene. Most actinomycetes produce bioactive compounds but the relationship between antibiotic production and morphology has not been studied in detail. Kitasatospora strains may be used as a model because they grow in submerged culture like those actinomycetes strains which produce bioactive compounds. The authors suggest further studies of the correlation between metabolic and morphological differentiation and the mechanism of the production of submerged spores. PMID- 10467660 TI - Hydrochloric acid treatment for rapid recovery of Shiga toxin-producing Escherichia coli O26, O111 and O157 from faeces, food and environmental samples. AB - We developed a hydrochloric acid treatment for the isolation of Shiga toxin producing Escherichia coli (STEC) O26:H11, O111:H- and O157:H7 strains from a variety of samples. After exposure to an equal volume of 1/8N HCl solution for 30 sec, the fecal suspensions and enrichment cultures were spread onto cefixime tellurite-sorbitol-MacConkey (CT-SMAC) agar. This HCl treatment increased the sensitivity for detection of STEC O26:H11, O111:H- and O157:H7 strains and decreased the growth of other microorganisms, from faecal samples and enrichment cultures of a variety of samples. This approach is an important economical and time-saving method to simplify and speed up isolation of STEC O26:H11, O111:H- and O157:H7 from a variety of samples. PMID- 10467661 TI - Persistence of Borrelia garinii and Borrelia afzelii in patients with Lyme arthritis. AB - We repeatedly detected DNA of Borrelia garinii or B. afzelii and Borrelia-like structures in the blood, joint fluid or in the synovium of 10 patients with Lyme arthritis by means of the polymerase chain reaction and immunoelectron microscopy at 2-4-month intervals in the course of two years. All samples were analyzed using primers which amplified the 16S rRNA gene sequence of Borrelia burgdorferi sensu lato and nucleotide sequences for the OspA gene. No cross hybridization occurred with DNA from human cells and with DNA from other bacteria. Capture and labelling with monoclonal antibodies of aggregated antigens, membranes and flagellae were evident in the blood of 7 patients, in 4 synovial membranes and 2 synovial fluids. Borreliae were found in blood capillaries, in collagen and in clusters surrounding inflammatory cells in the synovium of patients with recurrent infections who carried IgM and IgG antibodies to OspA and to 83 kDa core protein. After significant improvement for several weeks after treatment, arthritis recurred in six patients. Synoviocyte hyperplasia, inflammatory infiltration and concentric adventitial fibroplasia were seen in the synovium of the patients with persisting borreliae. Only two patients were infected with B. afzelii, the others with B. garinii. PMID- 10467662 TI - A nested RT-PCR for the detection of tick-borne encephalitis virus (TBEV) in ticks in natural foci. AB - We have developed a sensitive nested reverse-transcriptase polymerase chain reaction assay (n RT-PCR) for the detection of the tick-borne encephalitis virus (TBEV) RNA, especially in ticks. The primer pairs were selected from the 5' terminal noncoding region, a highly conserved part of the virus. The specificity was tested by computer homology searches of sequences as well as by the sequencing of the first and second amplificate, by Southern blot hybridization with a DIG-labelled oligonucleotide probe, and by restriction enzyme analysis. The method has proved to be very sensitive. The detection limit is about 20 fg of TBEV RNA per PCR run (25 microliters), or a single positive tick, i.e. (adult or nymph). The method can be used for comparative studies of the epidemiological situation, as well as for the screening of natural foci for the presence and circulation of TBEV or for the detection of TBEV-genome-sequences in clinical materials. PMID- 10467663 TI - Variations in the 16S rRNA gene sequence of Yersinia enterocolitica isolates influence the specificity of molecular identification systems. AB - Four identification systems were used to type Yersinia enterocolitica strain Y11 and Yersinia enterocolitica sensu strictoT. Two systems based on biochemical reaction patterns identified both strains as Yersinia enterocolitica. Two molecular assays targeting the 16S rRNA gene failed to identify either strain Y11 or the type strain. Therefore, both strains were typed by the classical taxonomical approach requiring a determination of the overall base composition and the base sequence similarity using hybridization. Again both strains were classified as Yersinia enterocolitica isolates. Consequently, the sequences of the 16S rRNA gene of both strains were determined and compared. The strains differed in a region where nucleotide changes between species of the genus Yersinia had been described earlier. These differences may explain the failure of the molecular assays to identify the strains. They also demonstrate an independent evolution of the 16S rRNA genes in the species Yersinia enterocolitica sensu stricto suggesting an amendment to the nomenclature to be used in the future. PMID- 10467664 TI - Extracellular product(s) of Staphylococcus aureus stimulate their own growth. AB - The effect of extracellular products obtained from culture supernatant of Staphylococcus aureus strain Cowan I on the bacterial growth was studied in a synthetic medium. Addition of the extracellular products to a fresh medium stimulated growth already after 2 h of incubation, with an approximately two-fold increase in cell density as compared to an unsupplemented medium, probably by promoting an initiation of growth accompanied by a reduction of the initial lag phase. The growth-stimulating effect was also monitored as an increase of adenosine triphosphate (ATP) in the bacterial culture during the different phases of growth. PMID- 10467665 TI - Determinants of pathogenicity of echovirus 9 in men: significance of a functional RGD-motif. AB - In this study, we investigated nine independent echovirus 9 isolates obtained from sick children in 1995. It is discovered that these isolates differ in respect to their pathogenicity for newborn mice indicating that the degree of human pathogenicity of an echovirus 9 variant does not necessarily correlate with mouse pathogenicity. Nevertheless, all virus variants are found to code for an RGD-motif within their VP1 protein. Hence, the RGD-motif and its highly conserved flanking regions are the conditio sine qua non, but, as expected, not sufficient for the mouse-pathogenic character. PMID- 10467666 TI - Adherence to and accumulation of S. epidermidis on different biomaterials due to extracellular slime production. In vitro comparison of a slime-producing strain (Rp 62 A) and its isogenic slime negative mutant (M7). AB - In an in vitro study, the bacterial adherence of a slime-producing strain (RP 62 A) was compared with its isogenic slime-negative mutant (M7). Standardized biomaterial discs were incubated under growth conditions in tryptic soy broth containing either strain RP 62 A or M7. After 24 h of incubation, the attached bacteria were removed by sonication and the colony-forming units were counted after plating of serial dilutions. We observed a significantly increased adherence and accumulation of the slime-producing strain (RP 62 A). In contrast to the slime negative mutant (M7) (p = 0.0001). The highest colony counts were found for the slime-producing strain on polyethylene and polymethylmethacrylate. The slime-negative mutant lacked the ability of accumulation. Our in-vitro results show the relevance of slime production by S. epidermidis for in-vitro colonisation of biomaterials, with a preference for polyethylene and polymethylmethacrylate. PMID- 10467668 TI - Onchocerciasis in Taraba State, Nigeria: clinical-epidemiological study of at risk males in Bakundi District. AB - An epidemiological study of 1366 adult males from fourteen communities of Bakundi district of Taraba State, Nigeria was carried out to establish the prevalence and intensity of onchocerciasis. The findings of the study showed that 45.2% of subjects were infected. Ten communities had infection rates in excess of 30%. Lizard skin and leopard skin were seen in 3.5% and 2.0%, respectively, and increased with increasing age. Hanging groin (2.6%) and scrotal enlargement (8.6%) were also observed. There was a good correlation between the infection rate on one hand and the presence of nodules (r = 0.85, P < 0.05), hanging groin (r = 0.56, P < 0.05) and scrotal enlargement (r = 0.62, P < 0.05) on the other. There was also a significant correlation between the intensity of infection on the one hand and lizard skin (r = 0.67, P < 0.05), leopard skin (r = 0.88, P < 0.05) and hanging groin (r = 0.82, P < 0.05) on the other. Those who worked in the fields (such as hunters, farmers and fishermen) had higher infection than others because of frequent contact with the vector. Extensive surveys of all suspected river valleys must be carried out to identify all those communities which will benefit from ivermectin treatment. PMID- 10467667 TI - False-positive beta-lactamase results with Staphylococcus lugdunensis in the Vitek AutoMicrobic system. AB - The Vitek AutoMicrobic system in combination with the Gram-positive susceptibility card detects beta-lactamase in staphylococci by utilizing penicillin as the substrate coupled with oxacillin as an inducer. The beta lactamase activity of 21 clinical isolates and two reference strains of Staphylococcus lugdunensis was determined with this automated system and compared with a liquid nitrocefin assay after induction with oxacillin. Eight (38%) clinical isolates and the reference strain ATCC 49576 of S. lugdunensis showed production of beta-lactamase in both tests. Thirteen (62%) clinical isolates and the type strain ATCC 43809 were nitrocefin-negative. The Vitek AutoMicrobic system reported false-positive beta-lactamase results for 9 of those 13 isolates and for the type strain of S. lugdunensis. Results for disk diffusion (ampicillin) were concordant with the nitrocefin assay. With one exception, the MICs for penicillin of the nitrocefin-negative strains were in the equivocal range of 0.06-0.12 mg/l according to NCCLS. However, none of the nitrocefin negative and Vitek-positive strains revealed any of the known staphylococcal genes for beta-lactamase as investigated by Southern hybridization, supporting the fact that false-positive beta-lactamase results may occur in the Vitek AutoMicrobic system. We conclude from our data that it may be justified to include S. lugdunensis in the quality control of Vitek cards containing beta lactamase tests. PMID- 10467669 TI - [Prenatal medicine--guarantee of a health child? Janus-head Medical advances]. PMID- 10467670 TI - [State of empirical research in psychotherapy]. AB - At present nearly 1200 psychotherapeutic treatments are offered on the market with all of them claiming to be helpful and effective. However many of them are not able to even fulfil minimal standards of scientifically substantiated evidence for effectiveness. Modern psychotherapy research has disclosed perspectives to adequately explore the complicated course of therapeutic process. The same way efforts for standardized judgement criteria of empirical studies offer the chance for meaningful evaluating the scientific quality of therapy studies. As shown by several meta-analyses the scientifically substantiated psychotherapy proved to be very effective in the treatment of mental disorders. PMID- 10467671 TI - [Preoperative clarification as change and burden]. PMID- 10467672 TI - [Psychosocial genetic counseling in patients with a high risk of breast cancer: experiences after 1300 meetings in the Amsterdam "Family Cancer Clinic"]. PMID- 10467673 TI - [Psychosocial impacts of surgical techniques]. PMID- 10467674 TI - [Coping strategies of high-risk pregnancies with threat of premature birtho]. PMID- 10467675 TI - [Sexuality in pregnancy: survey of the DFG project "Parenthood, couple relations and sexuality" and first preliminary findings]. PMID- 10467676 TI - [Holistic support of mother and child in the puerprium or: mother is more than "not a patient"]. PMID- 10467677 TI - [Possibilities and chances of psychological preparation for surgery: evaluation through gynecological patients]. AB - In this study the needs of 73 adult gynaecological patients concerning psychological preparation for surgery were examined. The patients could select between an information-providing and an anxiety-reducing psychological preparation mode, each including three components such as sensory information or guided imagery. The information were chosen by the majority of the patients (60.3%). When additionally asked to compose an (fictitious) individual psychological preparation out of the six components, the patients preferred different components depending on their age, preoperative anxiety and experiences with surgery. The results indicate that the psychological preoperative treatment should be individualized to match the needs of gynaecological patients. PMID- 10467678 TI - [Anxiety and coping behavior before and after radical gynecological surgery]. AB - Pretreatment phase-interactions between patient, family and medical staff around the time of diagnosis may have profound and long-term effects on quality of life. These interactions do not only influence the ability of the patient to come to terms with the diagnosis, but often set the tone for all future dealings with medical community. The imperatives for the medical staff in this process are to provide both: information and hope. In this context, anxiety and defense mechanisms--as protection of ego against anxiety--are psychoanalytic based constructs. The investigation of the interference between these constructs and information reflects the extraordinary background for an effective and continuing communication. These considerations require the care team to individualize their approach to each patient. PMID- 10467679 TI - [Treatment for infertility in HIV positive couples--psychosomatic and ethical considerations]. PMID- 10467680 TI - [The medicalization of women, The "myth of fragility"--the image of women in the advertising of hormonal replacement therapy]. PMID- 10467681 TI - [Predictors for refusal behavior of the mother towards her unborn and newborn child--first results of a prospective study]. PMID- 10467682 TI - [Prenatal diagnosis--fetal abnormalities--loss of pregnancy--a predetermined path?]. PMID- 10467683 TI - [Effect of stress and relaxation on the local immune system of the upper respiratory tract. A contribution to psychoneuroimmunology]. PMID- 10467684 TI - [The postmenopausal woman: relevant attitudes to health in aging]. PMID- 10467685 TI - [Working in balint groups--competence development the field of professional relations]. PMID- 10467686 TI - [In Process Citation] AB - The assessment of the activity of endometriosis provides a challenge during endoscopic procedures. Novel approaches can be developed in future based on more detailed understanding of the mechanisms of the pathogenesis of endometriosis. Angiogenic factors of local immunoreactions are important agents in the development and progression of the disease. Vascular endothelial growth factor (VEGF) is a potent inducer of vascular permeability and of the mitogenesis of endothelial cells. In endometriotic and endometrial tissues VEGF isoforms VEGF121, VEGF165, VEGF189 and VEGF206 are expressed. The amount of VEGF isoforms in the endometrium is regulated depending on the menstrual cycle. Macrophage recruitment by ectopic endometrial implants is an early step in the cascade of implantation and local peritoneal reaction. RANTES (Regulated upon Activation, normal T-cell-Expressed and Secreted) is a specific chemoattractant for monocytes and activated T-cells. Soluble products of activated macrophages play an important role in endometriosis associated infertility. The factors involved in angiogenesis and immune response are potential parameters to determine the activity of endometriotic implants and novel approaches for the treatment of endometriosis. PMID- 10467687 TI - [In Process Citation] AB - In a study 112 patients with histologically confirmed mostly recurrent endometriosis underwent a "three-step" therapy. Surgical removal of endometriosis implants was followed by a 6-months treatment with 3.75 mg of leuprorelin acetate depot as monthly subcutaneous injection and a second look laparoscopy with removal of residuals was performed. The follow-up period was up to 60 months (median 33.5 months). More than 50% of the patients had been pretreated by surgery and 30% by various medications. The r-AFS score pre- and postoperatively at the first- and post-therapeutic at the second look laparoscopy confirmed, that the primary operative intervention reduces the total r-AFS score by only 34%, whereas the combined approach reduces the score by 66%. These data of our own study could confirm, that in patients with symptomatic recurrent endometriosis the recurrence free time was dependent on the r-AFS score reached post therapeutically and this holds true for all stages. The higher the posttherapeutic r-AFS score the shorter the interval to recurrence. 91/112 (83.3%) patients complained infertility. 43/91 (47.3%) became pregnant during the follow-up period resulting in 36 newborns. Conception was spontaneously in more than 50% and stimulation programs were necessary in the others. Quality of life was regained in 54.9% and improvement of subjective conditions were reported in 52.9%. In our study we could confirm a clear relationship of recurrence of symptomatic endometriosis to the posttherapeutically achieved r-AFS score. Early aggressive therapy of active endometriosis is the optimal prevention of infertility caused by endometriosis. PMID- 10467688 TI - [In Process Citation] AB - Endometriosis and infertility are frequently associated. There is controversy in the literature about the causal correlation. There is no doubt, that progressed disease causes infertility by mechanical disturbances of the reproductive organs. Mild and moderate endometriosis however is discussed to cause functional infertility by changes of the peritoneal fluid, by endocrine and immunological dysfunction. Ovulation, ovum transport, fertilisation and implantation has been shown to be negatively influenced in patients with endometriosis. Some randomised prospective clinical studies however have demonstrated, that pregnancy rates after medical or surgical treatment of endometriosis are not improved compared with no treatment of the disease. The morphological data of this study may explain these conflicting data of the past: inactive, not proliferative endometriosis is not a cause of infertility, but in contrast active proliferating disease reduces fertility and needs sufficient treatment. PMID- 10467689 TI - [In Process Citation] AB - Endometriosis is a proliferative disease, which is often diagnosed with delay. Laparoscopy and histology are necessary for diagnosis. Different factors influence the success of therapy. Proliferative markers are very important for the differential diagnosis of active and inactive endometriosis. In a study it could be shown that endometriosis with high proliferative activity (high expression of Ki67) has a high response rate to GnRH-analogues treatment. Active endometriosis has to b distinguished to select an optimal treatment. A high proliferative activity of endometriosis should be treated with a hormonal surgical combination. PMID- 10467690 TI - [In Process Citation] AB - A total of 94 pre- or peri-menopausal patients from eight centres with intractable uterine bleedings and completed family planning were assessed as part of a clinical trial on the use of leuprorelin acetate monthly depot prior to endometrial ablation. The primary target criterion was flattening of the endometrium after two injections of 3.75 mg leuprorelin acetate depot with an interval of one month between injections. Endometrial thickness before the first injection was compared with the thickness prior to surgery (two weeks after the second injection). Mean endometrial flattening of 4.0 +/- 4.1 mm was found. Sufficient pretreatment was achieved in 91.3% of the patients with > 50% decrease. In terms of clinical response, the development of amenorrhoea, hypomenorrhoea or normal menstruation was assessed six weeks and still 83% after six months. In view of the good study results, hormone-suppressive pretreatment can be recommended prior to hysteroscopic endometrial ablation. Surgery should then take place about two weeks after the second injection. PMID- 10467691 TI - [In Process Citation] AB - GnRH agonist (GnRHa) administered for 6 months leads to an effective desensitisation of the pituitary and hypoestrogenism without exerting a particular effect on the whole metabolism. At the end of the first month's a suppression of the serum estradiol levels are achieved, the level of LH and FSH decline in the hypogonadotropic range. No negative influence on the lipid metabolism after administration of GnRH agonist has been observed. The balance of HDL/LDL does not change during the treatment. There were neither any negative changes in the liver metabolism, kidney function nor in the electrolyte values. In anaemic premenopausal women, for example due to serious menstrual problems, a normalisation of the haemoglobin concentration is obtainable already after a 12 week treatment. With regard to the hemostatis system a significant reduction of the procoagulant activity, fibrin turnover rate and a significant improvement of fibrinolytic activity can be observed under a GnRHa therapy. Although the use of GnRHa leads without doubt to a drastic reduction in the uterus blood flow there are no signs that this also leads to a change in the cerebral arteries blood flow. Menstrual bleeding occurs on average 3 months after the last injection of an GnRHa depot injection; with daily injection or nasal spray 3 to 4 weeks earlier. Theoretical considerations as well as the world-wide use as part of the infertility treatment--in some countries more than 90% of all IVF-cycles are performed using GnRH--,contradict the fact that GnRHa cause a teratogenic effect. Domineering undesirable side-effects during a treatment with GnRH can be traced back almost exclusively to the effective hormonal deprivation. In this context it is remarkable which percentage patients complain about trouble of this spectrum before GnRHa treatment is initiated. The chronicle reduction of the sexual hormone level leads without a doubt to a reduction of bone mineral density. The clinical relevance is furthermore a matter of controversial discussion. Prevention measures can be undertaken through an add-back therapy. This can also be of help in the case of vegetative side-effects caused by a decrease in sexual hormones. The question arises to what extent effective non hormonal add-back therapies are at disposal in the treatment of sexual hormone related malignant tumours. Also men with testosterone deprivation can suffer from distinctive hot flushes, sleeping disturbances and depression which requires some kind of relief in order to maintain an acceptable quality of life. PMID- 10467692 TI - [In Process Citation] AB - In a European, randomised Phase III trial, we studied ovarian ablation using a GnRH-agonist (leuprorelin acetate 3M depot, Takeda Pharma GmbH, Aachen) with CMF combination chemotherapy in pre- and peri-menopausal, hormone receptor-positive patients with involvement of 1-9 axillary lymph nodes. All planned 600 patients were recruited. We carried out an interim analysis of the hormonal effects and tolerability in 133 patients. The "castration effect" of the chemotherapy has previously been suspected but not systematically investigated. The hormonal effect should be an important aspect of adjuvant chemotherapy for node-positive breast cancer. For the first time we were able to demonstrate a pronounced estradiol-lowering effect of a "standard" chemotherapy regimen at least for the duration of the treatment. The side-effect profile of the two treatments in the interim analysis revealed marked advantages in favour of the GnRHa therapy. Currently no differences in efficacy are noted. However, sound conclusions can only be drawn after a far longer observation period, particularly after stratification according to the extent of node involvement and hormone receptor status. PMID- 10467693 TI - Molecular phylogenetics of DNA 5mC-methyltransferases. AB - We have identified a total of 88 members of the DNA-(cytosine-5) methyltransferase (5mC MTase) family whose sequences have been deposited in the databases. The results of a comparison of these sequences is presented in the form of an alignment-based phylogenetic tree and sequence logos for 10 conserved motifs. Phylogenetic analysis showed that members of the family aggregate into subfamilies which are usually consistent with their target specificity. However, it was also shown that similar target specificity does not necessarily imply close homology of the catalytic domain of MTases, which strongly supports the hypothesis that target recognition evolved independently of catalytic properties. This analysis also indicate that the 5mC MTase was present in the cenancestor (last common ancestor) of eubacteria, archaebacteria, and eukaryotes. The phylogeny of the 5mC MTases catalytic domain provides the basis for establishing the patterns of evolutionary change that characterize this family of proteins with conserved structural core and variable and mobile modules not directly involved in formation of the active site. PMID- 10467694 TI - Osmoregulation-dependent expression of Yersinia enterocolitica virulence factors. AB - The effects of hyperosmotic stress on expression of plasmid coded Yop and Yad A proteins--virulence factors of Y.enterocolitica serotype 0:9 were characterized. When cells were shifted to high osmolarity and cultured at 37 degrees C in medium without Ca2+ the production of Yops was inhibited. In contrast, the amount of Yad A protein was unaffected. Addition of glycine betaine to this culture alleviated the effect of high osmolarity. It was also found that hyperosmotic stress causes increased negative supercoiling of DNA in Y. enterocolitica 0:9. Changes in DNA supercoiling coincided with expression of Yop proteins. These results suggest that in high osmolarity the expression of yop genes may be regulated by DNA supercoiling. PMID- 10467695 TI - Methylobacterium extorquens strain P14, a new methylotrophic bacteria producing poly-beta-hydroxybutyrate (PHB). AB - Strain P14 of facultative methylotrophic bacteria that synthetisizes poly-beta hydroxybutyrate has been isolated. The cells are gram-negative motile rods with a polar flagellum. They do not form spores or capsules, but do have a caretenoid pigment. Predominant in the fatty acid composition of the cells is cis-vaccenic acid (cis 18:1: omega 7)--72%. In the phospholipid composition phosphatidylcholine predominates (45%), along with phosphatidylenthanoloamine (27%) and phosphatidylglycerol (17%). The main biquinone is Q-10; other ubiquinones (Q-8, Q-9, Q-11) are present in minor quantities. The cells accomplish the icl-variant of serine pathway. The GC content of DNA (Tm) is 65 mole%. A high level DNA-DNA homology with representatives of the genus Methylobacterium was observed. The strain has been identified as Methylobacterium extorquens strain P14. PMID- 10467696 TI - Identification of cytomegalovirus (CMV) infection by different laboratory methods in renal transplant recipients undergoing triple-drug immunosupressive treatment. AB - Early diagnosis of CMV infection is very important mainly in transplant recipients because CMV infection is a frequent complication after transplantation. In this work we compared different laboratory methods: ELISA (IgG, IgM), Western blot,shell vial, antigenemia assay (pp65), the immunofluorescent method with epithelial cells from urine (IF), DNA in leukocytes by PCR and DNA in leukocytes by hybridization (HCS) to estimate the most proper method for diagnosis of CMV in renal transplant recipients. This preliminary study showed that HCS, PCR and Western blot are sensitive methods for detecting CMV infection. Using HCS in quantitative variant we obtained a very good correlation between DNA load and clinical symptoms. PMID- 10467697 TI - Indigenous microflora responses to introduction of cyanogenic strains of Pseudomonas fluorescens into soil. AB - The effects of cyanogenic Pseudomonas fluorescens strains introduced into soil on the kinetic of colony formation and bacterial community structure were investigated. About 7.8 x 10(8) and 1.2 x 10(9) cfu per g dry soil of TA1 and B2 were added to the soil portions, respectively. The parameters of colony formation by heterotrophic soil bacteria were determined. The bacterial community structure and phenotypic diversity were studied using concept of r/K strategies and echophysiological index, respectively. The physiological state of indigenous heterotrophic bacteria and gram-negative group did not change under the influence of the cyanogenic strains introduced. Phenotypic diversity of the soil bacteria also did not change significantly. However, some short-term shifts in community structure of indigenous heterotrophic bacteria were noticed. This study shows that the introduction of great numbers of cyanogenic P. fluorescens strains could be safely used as potential agents in biological control of soil-born pathogens. PMID- 10467698 TI - Mitochondrial DNA variation in cattle of south China: origin and introgression. AB - Ten restriction endonucleases were used to investigate the mitochondrial DNA restriction fragment length polymorphism (mtDNA RFLP) of 11 native cattle breeds and one cultivated cattle breed in South China. Twenty-three restriction morphs were detected, which can be sorted into five haplotypes. A phylogenetic tree of the haplotypes was constructed by using the 'upgMa' method. Our study showed that haplotype I and II are identical to the zebu (Bos indicus) and taurine (Bos taurus) haplotypes, respectively. Zebu and taurine were the two major origins of cattle populations in South China, and the zebu probably had more influence on the native cattle population than taurine did. Haplotype III is identical to haplotype I of yak (Bos grunniens), which was only detected in the Diqing cattle breed. Haplotype IV was detected for the first time. This haplotype, found only in Dehong cattle, might be from an independent domestication event, probably from another Bos indicus population. Divergence of haplotypes I and IV occurred about 268,000-535,000 years ago, much earlier than the 10,000-year history of cattle husbandry. Our results also suggest a secondary introgession of mtDNA from yak to Diqing cattle. PMID- 10467699 TI - Microsatellite markers from a microdissected swine chromosome 6 genomic library. AB - To develop additional microsatellite (MS) markers in the region of the porcine skeletal muscle ryanodine receptor gene (RYR1), a microdissected genomic library was generated from the proximal half of the q arm of swine chromosome 6. Purified DNA was restriction enzyme-digested, ligated to oligonucleotide adaptors and amplified by PCR using primers complementary to the adaptor sequences. The purity of the amplified products and boundaries of the microdissected chromosomal region were verified by fluorescence in situ hybridization. (CA)n-containing sequences were then identified in a small insert genomic library generated from the PCR amplified microdissected DNA. Oligonucleotide primers were developed for the PCR amplification of 30 of the 46 (CA)n repeat-containing clones, which were subsequently used to amplify DNA isolated from unrelated pigs of different breeds to determine the informativeness of these MS markers. Twenty-two of these MS markers were genotyped on the University of Illinois Yorkshire x Meishan swine reference population. These 22 markers were all assigned within a 50.7-CM region of the swine chromosome 6 linkage map, indicating the specificity of the microdissected library. PMID- 10467700 TI - Genetic characterization of biodiversity in highly inbred chicken lines by microsatellite markers. AB - Forty-two microsatellite loci were analysed in 23 highly inbred chicken lines derived from Leghorn, Jungle Fowl, Fayoumi and Spanish breeds. Line-specific alleles among breeds and lines were detected. The band-sharing (BS) values were calculated and the proportion of shared alleles distances (Dps) were estimated. The BS values and Dps between sets of MHC-congenic lines ranged from 0.74 to 0.96, and 0.05-0.35, respectively. The BS values between each pair of noncongenic Leghorn lines were 0.32-0.97, and between Leghorn and exotic (Jungle Fowl, Fayoumi and Spanish) breeds were 0.03-0.55. The Dps between Fayoumi lines and other lines were much larger (0.66-1.34) than within Leghorns, and the Jungle Fowl breed had the largest distances with other lines (1.12-5.38). The phylogenetic consensus tree that was constructed grouped these 23 inbred chicken lines into four different clusters. These results are in accordance with the origin and breeding history of these inbred lines, which indicates that the use of microsatellites for the study of genetic biodiversity is accurate and reliable. In addition, the significance and value of inbred chicken lines in molecular genetic research is discussed. PMID- 10467701 TI - Satellite DNA polymorphisms and AFLP correlate with Bos indicus-taurus hybridization. AB - We describe satellite DNA variation that detects hybridization of Bos indicus (zebu or indicine cattle) and Bos taurus (taurine cattle) in African cattle populations. On Southern blots hybridized to a satellite III probe, relative intensities of Hinfl fragments correlated with the taurine-zebu composition in hybrid animals as deduced from AFLP genotyping of the same animals and previous data on microsatellite allele frequencies. Similar results were obtained by PCR RFLP analysis of a zebu-specific mutation in the repeat unit of satellite 1.711b. Analysis of individuals from 20 African cattle breeds indicate that the centromeric satellites of the sanga breeds are of the taurine type and that several East-African zebu breeds are hybrids between taurine and zebu. These satellite RFLP, or SFLP, markers provide a fast method to screen the genetic makeup of African cattle. PMID- 10467702 TI - Multicolour fluorescent detection and mapping of AFLP markers in chicken (Gallus domesticus). AB - We describe the mapping of amplified restriction fragment polymorphism (AFLP) markers in chicken (Gallus domesticus) using a multi-colour fluorescent detection system. DNA was used from a population consisting of four families with a total of 183 F2 individuals. The enzyme combination EcoRI/TaqI was used for double digestion, and fluorescently labelled fragments were analysed on an ABI PRISM 377 DNA sequencer. Polymorphic signals in the range of 50-500 bp were genotyped with the ABI PRISM Genotyper 2.0 software, which enabled the analysis of both dominant and incomplete dominant markers (with respect to AFLP, often referred to as codominant). In 19 sets consisting of 3 EcoRI/TaqI primer pair combinations each, a total of 475 polymorphic markers was detected. From these polymorphisms 344 markers could be mapped on the Wageningen linkage map. Fourteen markers were length polymorphisms of the same fragment and 28 markers Z-linked and uniformative; 64 AFLP markers appeared to be unlinked and 25 AFLP markers could not be accurately mapped on the basis of the genotyping results. The resulting AFLP/microsatellite linkage map is comprised of 33 linkage groups with a total of 835 loci. PMID- 10467703 TI - Catalase gene is associated with facial eczema disease resistance in sheep. AB - Facial eczema (FE) is a hepatogenous photosensitization disease of ruminant animals, particularly in sheep which vary widely in their susceptibility to the disease. The liver damage is caused by the mycotoxin, sporidesmin. There is evidence that the toxicity of sporidesmin is due to its ability to generate 'active oxygen' species. We evaluated the catalase gene, which encodes an enzyme with antioxidant functions, as a candidate for determining the susceptibility of sheep to the disease. Two microsatellite markers, OarSHP3 and OarSHP4, which flank the sheep catalase gene, were isolated from a Yeast Artificial Chromosome (YAC) clone. These markers mapped the catalase locus by linkage to ovine chromosome 15. Eleven informative markers spaced throughout chromosome 15, inclusive of the catalase marker OarSHP4, gave no significant linkage with the disease traits when analysed in four outcross resource pedigrees. However, OarSHP3 and OarSHP4 allele frequencies showed significant differences between FE resistant and susceptible selection-lines. Comparison of sequences of catalase cDNAs from sheep of resistant and susceptible lines showed only two silent mutations. A single nucleotide polymorphisms (KP1) in exon 6 of the catalase gene also showed significant differences in allele frequencies between the selection lines. The lack of evidence for linkage in outcross pedigrees, but the significant association in the genetic lines, implies that catalase is involved in determining the susceptibility of sheep to facial eczema, and that the candidate gene's effect is probably recessive or minor. PMID- 10467704 TI - An MspI polymorphism at the bovine growth hormone (bGH) gene is linked to a locus affecting milk protein percentage. AB - SSCP analysis of the bovine growth hormone (bGH) gene in Israel Holstein dairy cattle uncovered five intragenic haplotypes, denoted A to E. Of these, Haplotype E differed from the others at six fragments; one of which corresponded to the polymorphic MspI site in intron III, at which haplotype E carried the disabled MspI (-) allele. Haplotype E was observed in a single sire only, carrying haplotype A as the second bGH allele. In 523 daughters of this sire genotyped for the MspI polymorphism, heterozygous (+/-) as compared to homozygous (+/+) daughters, showed a significant increasing effect on protein percentage and kg protein per year; and a decreasing effect (P < 0.10) on milk somatic cell counts (MSSC). None of the daughters were homozygous (-/-), indicating that the frequency of this allele in the general population was essentially zero. Calculated skewness (g1) values for the two daughter groups differed significantly with (+/-) daughters showing negative skewness (in the direction of lower protein percentage), and (+/+) daughters positive skewness (in the direction of higher protein percentage). The direction of skewness in each group is indicative of the presence of a QTL having an increasing effect on milk protein percentage in coupling linkage with the MspI (-) allele in this sire, but at some distance from it. Maximum likelihood estimates of the proportion of recombination (r) between the putative QTL and bGH, and the allele substitution effect at the QTL (d), were r = 0.33, a = 0.07% protein, with standard errors 0.058 and 0.009% protein, respectively. PMID- 10467705 TI - A novel integral membrane protein is differentially expressed in the chick growth plate and maps to chromosome 1. AB - The growth plate is a specialised region of cartilage located at the growing ends of long bones in higher vertebrates. It is responsible for longitudinal bone growth and is under the control of many local and systemic factors. The growth plate consists of an orderly arrangement of small proliferative and larger mature hypertrophic chondrocytes. This paper describes the isolation by differential display of a 988-bp cDNA fragment derived from a transcript that is more highly expressed in proliferating rather than hypertrophic chondrocytes of the chick growth plate. Using 3' RACE, a further 939 bp of cDNA sequence was obtained. The 1.9 kb sequence contains a 924-bp open reading frame encoding an unknown 308 amino acid protein. This protein has a putative transmembrane domain near its N terminus and three dileucine motifs at its carboxy tail. This gene was expressed in all other tissues examined. A polymorphism was identified by SSCP analysis and the gene was mapped to the centromeric region of the short arm of chicken chromosome 1, close to the locus for autosomal dwarfism. PMID- 10467706 TI - Physical assignments of human chromosome 13 genes on pig chromosome 11 demonstrate extensive synteny and gene order conservation between pig and human. AB - Previous mapping between the human and pig genomes suggested extensive conservation of human chromosome 13 (HSA13) to pig chromosome 11 (SSC11). The objectives of this study were comparative gene mapping of pig homologs of HSA13 genes and examining gene order within this conserved synteny group by physical assignment of each locus. A detailed HSA13 to SSC11 comparison was chosen since the comparative gene map is not well developed for these chromosomes and a rearranged gene order within conserved synteny groups was observed from the comparison between HSA13 and bovine chromosome 12 (BTA12). Heterologous primers for PCR were designed and used to amplify pig homologous fragments. The pig fragments were sequenced to confirm the homology. Six pig STSs (FLT1, ESD, RB1, HTR2A, EDNRB, and F10) were physically mapped using a somatic cell hybrid panel to SSC11, and fluorescent in situ hybridization (FISH) mapping was also applied to improve map resolution and determine gene order. Results from this study increase the comparative information available on SSC11 and suggest a conserved gene order on SSC11 and HSA13, in contrast to human:bovine comparisons of this syntenic group. PMID- 10467707 TI - The L-gulono-gamma-lactone oxidase gene (GULO) which is a candidate for vitamin C deficiency in pigs maps to chromosome 14. AB - Vitamin C deficient pigs, when fed a diet lacking L-ascorbic acid (AscA), manifest deformity of the legs, multiple fractures, osteoporosis, growth retardation and haemorrhagic tendencies. This trait was shown by others to be controlled by a single autosomal recessive allele designated as od (osteogenic disorder). The inability of AscA biosynthesis in primates and guinea pigs that exhibit similar symptoms, when they are not supplemented with AscA in the food, was traced to the lack of L-gulono-gamma-lactone oxidase, which catalyzes the terminal step in the biosynthesis of AscA. The non-functional GULOP was mapped to human chromosome 8p21 that corresponds to an evolutionarily conserved segment on either porcine chromosome 4 (SSC4) or 14 (SSC14). We investigated linkage between OD and SSC4- and 14-specific microsatellite loci in order to map the OD locus. Twenty-seven informative meioses in families from one sire and three dams revealed linkage of od with microsatellites SW857 and S0089, located in the subcentromeric region of SSC14. We isolated part of the GULO gene of the pig by screening a porcine genomic library using a pig GULO cDNA as a probe, and mapped it to SSC14q14 by fluorescence in situ hybridization (FISH). Thus, the porcine GULO gene is both a good physiological and positional candidate gene for vitamin C deficiency in pigs. PMID- 10467708 TI - Effect of genetic variability of the porcine pituitary-specific transcription factor (PIT-1) on carcas traits in pigs. AB - Pituitary transcription factor (PIT-1) has been shown to be a positive regulatory factor of growth hormone, prolactin, and thyrotrophin-beta-subunit (TSH-beta) in the mammalian pituitary. Therefore, the gene encoding PIT-1 (POU1F1) was chosen as a candidate gene to investigate its association with growth and carcass traits in pigs. The purpose of this study was to analyse porcine POU1F1 genetic variability in populations of Large White and Large White x landrace pigs, by using PCR-RFLP analysis and to determine its possible associations with two carcass traits (backfat and percentage of lean content). Two different POU1F1-PCR RFLP (POU1F1/RsaI and POU1F1/MspI) tests were applied to genomic DNA isolated from porcine blood (120 pigs) and hair roots (10 pigs). The present results clearly indicated that the MspI DD genotype was the fattest compared with both other genotypes (CC, CD) in analyzed swine population. For POU1F1/RsaI polymorphism no significance differences were seen for lean-to-fat ratio. PMID- 10467709 TI - Three PCR-RFLPs in the kappa-casein (CSN3) gene in American bison. PMID- 10467710 TI - A five allele single strand conformational polymorphism in the third intron of the ovine CD3D gene. PMID- 10467711 TI - Single strand conformational polymorphisms (SSCPs) in the ovine IL1A and IL6 genes. PMID- 10467712 TI - Five multiplexed microsatellite loci for rapid response run identification of California's endangered winter chinook salmon. PMID- 10467713 TI - Equine dinucleotide repeat loci COR041-COR060. PMID- 10467714 TI - Two highly polymorphic microsatellites within the porcine ryanodine receptor 3 gene (RYR3). PMID- 10467715 TI - A multiplex PCR-SSCP test to genotype bovine beta-casein alleles A1, A2, A3, B, and C. PMID- 10467716 TI - Linkage assignments for 14 SRC bovine microsatellites. PMID- 10467717 TI - Overview of endocrine disruptor research activity in the United States. AB - The issue of whether environmental contaminants are inducing adverse health effects in humans and wildlife via interaction with endocrine systems has gained increasing interest during the 1990s. Endocrine disruption is one of the highest priority research topics for the US EPA, and a detailed research strategy has been developed to guide the placement of resources over the next several years. To address the deficiency of testing guidelines in detecting and characterizing damage mediated by interaction with the endocrine system, EPA has issued new multi generation testing guidelines. The new endpoints for monitoring pubertal development, semen quality, and estrous cyclicity will better enable determination of the affected sex, target organ, and life stage following exposure throughout the life cycle. Another major area of effort within EPA is the development of an endocrine disruptor screening program in response to passage of the Food Quality Protection Act of 1996. The current status of these efforts is described. On the federal level, endocrine disruption is one of the five priority research areas for the Committee on the Environment and Natural Resources (CENR) within the Executive Office of President. The CENR has developed a framework to assess research needs for endocrine disruptors, inventoried existing efforts of the federal government (nearly 400 projects were identified as active in FY96), and prioritized additional research needs based upon the needs and gaps in current efforts. It is clear that a great deal of research is underway to clarify the validity of the endocrine disruptor hypothesis and to determine the breadth of chemicals that pose a risk to the endocrine system. The degree of forward research planning and coordination across many organizations should ensure that sufficient data will be available within the next few years to allow a rigorous weight of evidence evaluation that is needed to bring together diverse types of information to make informed decisions regarding risks to humans and wildlife. PMID- 10467718 TI - Ecological effects of endocrine disruption: current evidence and research priorities. AB - An overview is presented of the main evidence for endocrine disruption in wildlife, focusing on reproductive effects. While there are a few clear examples of endocrine disruptive effects resulting from environmental chemical exposure, in most cases a causal link between the observed abnormalities and chemical exposure has not been established. In other cases there appears to be a link but the specific chemicals responsible for the observed effects, and the mechanisms involved, remain to be elucidated. Also it is largely unknown whether or not observed changes in individual animals lead to population-level effects. Priority research projects for the UK, ultimately aimed at determining the population level significance of endocrine disruption, are described. PMID- 10467719 TI - Effects of organochlorine xenobiotics on human spermatozoa. AB - The organochlorine insecticide lindane is a widely distributed environmental pollutant belonging to the growing family of endocrine disrupting chemicals (EDCs). Lindane intercalates into the sperm membrane and alters the molecular dynamics of the bilayer. In the present paper, preliminary data are reported showing that doses of lindane as low as those found in the female genital tract secretions inhibit the sperm cytological responsiveness to progesterone, the physiological agonist which stimulates the onset of acrosome reactions at the site of fertilization. The hypothesis is put forth that even background levels of lindane may exert antifertility effects independently on the health status of either the male and female reproductive organs. PMID- 10467720 TI - Specific issues in health risk assessment of endocrine disrupting chemicals and international activities. AB - Specific issues in health risk assessment of EDCs and correlated international activities thereof are discussed. Risk characterization is a synthesis of all information including hazard assessment, dose-response relationship, and exposure information to identify clearly the strengths and weaknesses of the database, the criteria applied to evaluation and validation of all aspects of methodology, and the conclusions reached from the review of scientific information. In the case of EDCs, new models need to be developed taking into account possible new kinds of information, e.g., effects of EDCs on gene activation in response to hormonal challenge or effects on receptor expression. Such models should also account for homeostatic adaptive responses and consider the possibility of having windows of exposure for given effects. Work to compile and harmonize the definitions and terms appropriate to endocrine disruption will be conducted within the joint IPCS/OECD project on harmonization of risk/hazard assessment terminology reviews. The ICPS is the process of preparing a "State of the Science" report, and is implementing a global inventory of ongoing research on ECDs. PMID- 10467721 TI - Behavioural effects of endocrine disrupting chemicals on laboratory rodents: statistical methodologies and an application concerning developmental PCB exposure. AB - Appropriate behavioural tests and adequate statistical tools may help to establish the ED properties of a given compound by pointing out the alterations of selected behavioural endpoints. Frequently, laboratory collected data consist of frequencies and/or durations of specific items, and the analysis of variance (ANOVA) technique is performed to assess whether the investigated factors affect these behavioural endpoints. Moreover, when numerous aspects of behaviour are investigated simultaneously, Principal Component Analysis (PCA), a multivariate technique, may be very useful to reduce the overwhelming number of correlated original variables to a few orthogonal artificial variables (factors). Continuous Time Markov Chain (CTMC) models may be applied to analyse the time structure of a behavioural pattern when data consist of sequences of events and the time points at which they occur. Moreover, the Cox Proportional Hazard Model, a methodology originally developed for the analysis of failure time data, may help to evidence the effects of a given treatment on behavioural sequences when the assumptions of CTMC models are not fully satisfied. Analyses on data from mice of the outbred CD 1 strain (controls in a study of toxicity and exposed to PCB during development) are presented as examples to show how adequate statistical analyses and appropriate behavioural tests may reveal relevant effect of treatments otherwise not easily detected. PMID- 10467722 TI - Nondestructive biomarkers of exposure to endocrine disrupting chemicals in endangered species of wildlife. AB - This paper explores the problem of endocrine disrupting chemicals (EDCs) from the ecotoxicological point of view, focusing on nondestructive biomarkers of exposure to EDCs for risk assessment of endangered species of wildlife. Several EDCs, such as polyhalogenated aromatic hydrocarbons and toxic metals, tend to be biomagnified in the terrestrial and particularly the marine food chains. Top predators tend to accumulate high concentrations of these contaminants which places them in a situation of high toxicological risk. Hence, there is a need to develop nondestructive techniques, such as nondestructive biomarkers, for hazard assessment, protection, and conservation of endangered species exposed to EDCs. The biological materials proposed for this approach (for example blood, faeces, fur, skin biopsy specimens) are easily obtained with minimal stress for individuals and populations. Some validation data are reported on porphyrins in sea bird excreta (Larus dominicanus, Phalacrocorax olivaceus, Pelecanus occidentalis thagus), as nondestructive biomarkers of exposure to organochlorines, and on benzopyrene monooxygenase activities in marine mammal skin biopsy specimens (Stenella coeruleoalba, Balaenoptera physalus), as early indicators of exposure to p,p'-DDE and other endocrine disrupting organochlorines. PMID- 10467723 TI - Dilemmas facing regulatory and advisory bodies dealing with conflicting results. AB - Regulatory and advisory bodies have to take into account the possibility that conventional toxicological screening and testing methods may be failing to pick up relevant effects, with the implication that testing may need to go down to much lower exposure levels and use specifically designed studies. In the paper, attention is focused on the problems of understanding endocrine disrupting activity. The shape of the dose-response curve for endocrine disrupters may differ from that normally associated with toxic chemicals (the monotonic sigmoid curve). Thus, prediction of both the nature of the effects of potential endocrine disrupters and the intensity of those effects at different doses may be equally difficult. Endocrine effects are not currently amenable to analysis by a single apical assay. While existing toxicological tests can pick up many end-organ and functional effects that are relevant to endocrine disturbances, they are generally poor at exploring apparently subtle effects such as disturbances of brain endocrine homeostatic mechanisms which may be critical for normal development. PMID- 10467724 TI - Problems in testing and risk assessment of endocrine disrupting chemicals with regard to developmental toxicology. AB - Endocrine disrupting chemicals (EDCs) may affect mammalian development either indirectly (by impairing implantation, placental development, lactation, etc.) or directly, altering the maturation of target tissues. Current regulatory tests for reproductive/developmental toxicity should be carefully evaluated with regard to risk assessment of EDCs, considering hazard identification (are relevant endpoints being assessed?) and dose-response assessment (are sensitive NOEL/dose response curves being provided?). Many in vitro and in vivo assays for sex steroid disruption are available; provided that the metabolic capacities of the assays are defined, they could be integrated in a sensitive battery for early detection of steroid-disrupting potentials. The screening battery should address further regulatory in vivo tests (e.g. what specific parameters have to be investigated). As regards dose-response, qualitative differences may be observed between lower and higher exposures, showing primary hormone-related effects and frank embryotoxicity, respectively. Other problems concern (a) the identification of critical developmental windows, according to hormone concentrations and/or receptor levels in the developing target tissues; (b) the potential for interactions between chemicals with common mechanism/target (e.g. xenoestrogens); (c) most important, besides sex steroids more attention should be given to other mechanisms of endocrine disruption, e.g., thyroid effects, which can be highly relevant to prenatal and postnatal development. PMID- 10467725 TI - Environmental and biological monitoring of endocrine disrupting chemicals. AB - Trends toward an increase of adverse health effects on reproductive organs have been reviewed. An urgent need has been recognised to establish validated in vivo and in vitro screening assays to test for hormonal activities of chemicals. Relevant existing OECD guidelines have been reviewed and enhancements of some of these have been identified, mainly to test for estrogenic and androgenic activity of chemicals. The problems connected to monitoring activities are outlined, particularly for ambient and biological monitoring. Indeed, the problem of assessing human exposure to endocrine disrupting chemicals through environmental chemical analysis tends to a very high level of complexity. This has been illustrated through the example of one single subclass of endocrine disrupting compounds (EDCs), the organohalogen compounds. Valid biological markers are also needed to be effectively used in epidemiological studies and risk assessment. A multidisciplinary approach and the collaboration among experts in the field of clinical biochemistry, toxicology, and epidemiology is required. PMID- 10467726 TI - Rapid treatment of water contamined with atrazine and parathion with zero-valent iron. AB - The utility of fine-grained iron metal in the remediation of water contamined with Atrazine and Parathion was investigated. Batch procedures under water treatment conditions (ambient temperature and pH of approximately 7) indicated that these pesticides degrade rapidly in the presence of iron powder (40-60 mesh, 40 g/l). The decline in the concentration of pesticide was monitored by HPLC. Experiments with unbuffered solutions showed a steady increase in pH values during the reactions. Therefore, experiments were run in buffered solutions. Different buffered solutions resulted in different degradation rates indicating that the buffer plays an important role in enhancing the degradation process. Tests were also performed on an industrial effluent solution containing a variety of pesticides. Although the products of degradation were not characterized, our HPLC results indicated the disappearance of all the parent pollutants. PMID- 10467727 TI - Effects of polydiallyldimethyl ammonium chloride coagulant on formation of chlorinated by products in drinking water. AB - The objectives of this research work was to evaluate the reduction of THM precursors by cationic p-DADMAC and determine the correlations between the chlorine demand and trihalomethane formation in the presence of electrolyte solutions and ambient light. The chlorine demand was found to be significantly reduced provided that the H2SO4 electrolyte was fed to the sample solutions. The amount of CHCl3 formation was also decreased when the Na2SO4 electrolyte was introduced in spite of the levels of light intensity. The p-DADMAC can not only effectively remove the turbidity but also reduce the formation of CHCl3. The optimum dosage of p-DADMAC for reducing the turbidity, TOC and CHCl3 in the humic acid and source water samples was determined and depended upon the nature of organics. PMID- 10467728 TI - ATM: the product of the gene mutated in ataxia-telangiectasia. AB - Ataxia-telangiectasia mutated (ATM) is the product of the gene mutated in the human genetic disorder ataxia-telangeictasia (A-T). It is a 370 kDa protein that is a member of the phosphatidyl inositol 3-kinases superfamily. A-T cells and those derived from Atm-/- mice are characterized by hypersensitivity to ionizing radiation and defective cell cycle checkpoints. Defects are observed at all cell cycle checkpoints in A-T cells post-irradiation including the G1/S interface where ATM plays an important role in the activation of the tumour suppressor gene product p53. Activation leads to the induction of p21/WAF1, inhibition of cyclin dependent kinase activity, failure to phosphorylate key substrates such as the retinoblastoma protein and consequently G1 arrest. ATM also plays an important role in the regulation and surveillance of meiotic progression. Absence of ATM gives rise to a spectrum of defects including immunodeficiency, neurodegeneration, radiosensitivity and cancer predisposition. It is clear that a better definition of the role of ATM in DNA damage recognition, cell cycle control and cell signalling may assist in the treatment of the progressive neurodegeneration in this syndrome. PMID- 10467729 TI - Fibrinogen. AB - Fibrinogen is a blood-borne glycoprotein comprised of three pairs of nonidentical polypeptide chains. Following vascular injury, fibrinogen is cleaved by thrombin to form fibrin which is the most abundant component of blood clots. As well as controlling blood loss at sites of tissue damage, other properties of fibrinogen have recently been discovered. For example, various cleavage products of fibrinogen and fibrin, released during coagulation and fibrinolysis, respectively, regulate cell adhesion and spreading, display vasoconstrictor and chemotactic activities, and are mitogens for several cell types including fibroblasts, endothelial and smooth muscle cells. Current research aims to define the bioactive fibrinogen molecule moieties and cellular receptors involved in these processes. Future studies may provide us with new opportunities to develop agents which are useful in promoting tissue repair or conversely in inhibiting fibrosis in inflammatory and fibroproliferative diseases where endothelial cell damage or chronic leakage of blood proteins is a feature. PMID- 10467730 TI - Aminopeptidase B (EC 3.4.11.6). AB - Aminopeptidase B (EC 3.4.11.6) is a Zn(2+)-dependent exopeptidase which selectively removes arginine and/or lysine residues from the NH2-terminus of several peptide substrates including Arg0-Leu-enkephalin, Arg0-Met-enkephalin and Arg-1-Lys0-somatostatin-14. Analysis of its primary structure showed that aminopeptidase-B is structurally related to leukotriene A4 hydrolase, an important enzyme of the arachidonic acid pathway. This structural relationship is further supported by the capacity of aminopeptidase-B to hydrolyse leukotriene A4. Aminopeptidase-B is widely distributed in a number of tissues, including endocrine and non-endocrine cells. Moreover, in rat PC12 pheochromocytoma cells, the enzyme is secreted and associated with the external face of the plasma membrane. Together these data strongly argue in favour of a role of this bi functional enzyme in the final stages of precursor processing mechanisms occurring either in the secretory pathway, at the plasma membrane, or at both locations. PMID- 10467731 TI - Quantitation of oxidative damage to tissue proteins. AB - Active oxygen species are thought to be involved in many physiological and pathological processes and are known to oxidatively modify DNA, lipids and proteins. One such modification is the addition of carbonyl groups to amino acid residues in proteins. The number of carbonyl groups on proteins can be quantitated spectrophotometrically using 2,4-dinitrophenylhydrazine (DNPH). The DNPH assay described in the literature was found to be unreliable in samples containing high amounts of chromophore (e.g. hemoglobin, myoglobin, retinoids). By using an HCl-acetone wash, hemes from the chromophores could be extracted, enabling the determination of carbonyl content to be made even in highly colored tissue extracts. Residual DNPH, which was also found to interfere with the assay, was removed by additional washes with trichloroacetic acid and ethanol ethylacetate. These improvements are known to remove lipids, do not lengthen the time required to do the assay, permit quantification of carbonyl content in 1-4 mg protein from a variety of tissue types and provide a sensitive and reliable method for assessing oxidative damage to tissue proteins. PMID- 10467732 TI - Renaturation of rabbit liver aminoacyl-tRNA synthetases by 80S ribosomes. AB - Protein biosynthesis machinery is thought to be mostly compartmentalised within the mammalian cell, involving direct interactions between different components of the translation apparatus. The present research concerns the functional meaning of the interaction between the rabbit liver aminoacyl-tRNA synthetases and 80S ribosomes. We have shown that rabbit liver 80S ribosomes are able to enhance the activity of leucyl-tRNA synthetase, which is a component of high-molecular weight aminoacyl-tRNA synthetase complex, and phenylalanyl-tRNA synthetase not associated within this complex. The ribosomes increase the initial rate of both the total reaction of tRNA aminoacylation and the first step of this reaction, the formation of leucyladenylate. Moreover, a positive cooperativity of the tRNA interaction with two binding sites of leucyl-tRNA synthetase is also increased in the presence of highly purified 80S ribosomes. The effect of 80S ribosomes on partly denatured leucyl-tRNA synthetase and phenylalanyl-tRNA synthetase and the protection by 80S ribosomes of both enzymes against inactivation indicate a refolding and stabilising capacity of the ribosomes. It is concluded that the interaction of aminoacyl-tRNA synthetases and 80S ribosomes is important for the maintenance of an active conformation of the enzymes. PMID- 10467733 TI - Modeling the Leigh syndrome nt8993 T-->C mutation in Escherichia coli F1F0 ATP synthase. AB - The mutations in human mitochondrial DNA at nt8993 are associated with a range of neuromuscular disorders. One mutation encodes a proline in place of a leucine conserved in all animal mitochondrial ATPase-6 subunits and bacterial a subunits of F1F0 ATP synthases. This conserved site is leu-156 and leu-207 in humans and Escherichia coli, respectively. An aleu-207-->pro substitution mutation has been constructed in the E. coli F1F0 ATP synthase in order to model the biochemical basis of the human disease mutation. The phenotype of the aleu-207-->pro substitution has been compared to that of the previously studied aleu-207-->arg substitution (Hartzog and Cain, 1993, Journal of Biological Chemistry 268, 12250 12252). The leu-207-->pro mutation resulted in approximately a 35% decrease in the number of intact enzyme complexes as determined by N, N' dicyclohexylcarbodiimide-sensitive membrane associated ATP hydrolysis activity and western analysis using an anti-a subunit antibody. A 75% reduction in the efficiency of proton translocation through F1F0 ATP synthase was observed in ATP driven proton pumping assays. Interestingly, the loss in F1F0 ATP synthase activity resulting from the leu-207-->pro substitution was markedly less dramatic than had been observed for the leu-207-->arg mutation studied earlier. By analogy, the human enzyme may also be affected by the leu-156-->pro substitution to a lesser extent than the leu-156-->arg substitution, and this would account for the milder clinical manifestations of the human leu-156-->pro disease mutations. PMID- 10467734 TI - Aldehyde dehydrogenase from rat intestinal mucosa: purification and characterization of an isozyme with high affinity for gamma-aminobutyraldehyde. AB - In rat adrenal gland and gastric mucosa putrescine is efficiently oxidized to GABA via gamma-aminobutyraldehyde (ABAL) by action of diamine oxidase and aldehyde dehydrogenase. Having turned our attention on the rat intestinal mucosa, where putrescine uptake and diamine oxidase are active, we have purified and characterized an aldehyde dehydrogenase optimally active on gamma aminobutyraldehyde. A dimer with a subunit molecular weight of 52,000, the native enzyme binds ABAL and NAD+ with high affinity: at pH 7.4, Km values are equal to 18 and 14 microM, respectively. Affinity for betaine aldehyde is much lower (Km = 285 microM), but the efficiency is equally good, thanks to a high value of V. Unaffected by disulfiram and Mg2+, the enzyme is activated by high NAD+ concentrations (Vnn = 1.6 x Vn) and is competitively inhibited by NADH. According to the best fitting model, the dimeric enzyme only binds one NADH and the mixed complex enzyme-NAD(+)-NADH is inactive. The increase of activity promoted by NAD+ can therefore be ascribed to an allosteric effect, rather than to the activation of a second reaction center. Highly stable at pH 6.8 in the presence of dithiothreitol and high phosphate concentrations, ABALDH is inactivated by ion exchange resins and by cationic buffers. Our results show that the enzyme can be effectively involved in the metabolism of biogenic amines and, with a K(m) for ABAL lower than 20 microM, in the synthesis of GABA. PMID- 10467735 TI - A rapid method for the purification of glucose-6-phosphate dehydrogenase from bovine lens. AB - This paper describes a simple and rapid method for the purification of glucose-6 phosphate dehydrogenase from bovine lens, together with analysis of the kinetic behaviour and some properties of the enzyme. The purification consisted of two steps, 2',5'-ADP-Sepharose 4B affinity chromatography and DEAE Sepharose Fast Flow ion exchange chromatography in procedure which took two working days. The enzyme was obtained with a yield of 13.7% and had a specific activity of 2.64 U/mg protein. The overall purification was about 19,700-fold. The molecular weight of the enzyme was found to be 62 +/- 3 kDa by Sephadex G-200 gel filtration chromatography. A protein band corresponding to a molecular weight of 69.2 +/- 3.2 kDa was obtained on SDS polyacrylamide slab gel electrophoresis. On chromatofocusing, lens glucose-6-phosphate dehydrogenase gave a single peak at pI 5.14. The activation energy of the reaction catalyzed by the enzyme was calculated from Arrhenius plot as Ea = 5.88 kcal/mol. The pH versus velocity curve had two peaks at pH 7.7 and 9.6. By the double-reciprocal plots and the product inhibition studies, it was shown that the enzyme follows 'Ordered Bi Bi' sequential kinetics. From the graphical and statistical analyses, KmNADP+, KmG-6 P, KiNADPH, Ki6-PGA were estimated to be 0.008 +/- 0.002, 0.035 +/- 0.013, 0.173 +/- 0.007 and 1.771 +/- 0.160 mM, respectively. The observed kinetic behaviour of glucose-6-phosphate dehydrogenase from bovine lens was in accordance with the enzyme from other sources. PMID- 10467736 TI - Increased adhesion of lymphoid cells to glycated proteins. AB - BACKGROUND AND AIMS: The advanced glycation end-products are involved in the pathogenesis of vascular damages and other clinical complications in diabetic patients. The aim of this study was to investigate the adhesion of lymphoid cells to nonenzymatically glycated proteins in comparison with the unmodified substances. METHODS: Two cell lines (monocyte-macrophage line U937 and the T-cell line Jurkat) were used throughout the experiments. The cells were left to adhere to nonenzymatically glycated and native proteins coated on a 96-well flat-bottom plates and the cellular adhesion was registered as absorption at 550 nm following the method described by Ivanov and Kyurkchiev [G. Ivanov, S. Kyurkchiev, Effect of advanced glycosylation end-products on the activity of integrins expressed on U937 cells, Hum. Immunol. 59 (1998) 325-330.]. RESULTS: It was found that the monocytes had increased adhesion to nonenzymatically glycated proteins such as collagen, fibronectin and bovine serum albumin, whereas the T-cells had increased adhesion to the glycated collagen and bovine serum albumin but reduced adhesion to advanced glycated fibronectin. Experiments with different stimulating agents showed that phorbol-myriastate, acetate (A550 = 0.672 +/- 0.068, S.E.M., n = 40), glucose (A550 = 0.593 +/- 0.051, S.E.M., n = 40) and TNF-alpha (A550 = 0.580 +/- 0.042, S.E.M., n = 40) increased the adhesion of U937 cells to advanced glycated bovine serum albumin in comparison with the adhesion of the untreated cells (A550 = 0.260 +/- 0.046, S.E.M., n = 40). This is probably due to an upregulation of the expression or the activity of the receptors for the advanced glycation end products. CONCLUSION: Based on the results obtained it is concluded that the receptors for nonenzymatically glycated proteins expressed on the surface of lymphoid cells could act also as cell adhesion molecules. PMID- 10467737 TI - Membrane protein structure determination by solid state NMR. PMID- 10467738 TI - Biosynthesis of polyketides (other than actinomycete macrolides). PMID- 10467739 TI - Excitatory amino acids. PMID- 10467740 TI - Recent advances in chemical ecology. PMID- 10467741 TI - The biosynthesis of shikimate metabolites. PMID- 10467743 TI - Laterality of expression in portraiture: putting your best cheek forward. AB - Portraits, both photographic and painted, are often produced with more of one side of the face showing than the other. Typically, the left side of the face is overrepresented, with the head turned slightly to the sitter's right. This leftward bias is weaker for painted male portraits and non-existent for portraits of scientists from the Royal Society. What mechanism might account for this bias? Examination of portraits painted by left- and right-handers and of self-portraits suggests that the bias is not determined by a mechanical preference of the artist or by the viewer's aesthetics. The leftward bias seems to be determined by the sitters and their desire to display the left side of their face, which is controlled by the emotive, right cerebral hemisphere. When we asked people to portray as much emotion as possible when posing for a family portrait, they tended to present the left side of their face. When asked to pose as scientists and avoid portraying emotion, participants tended to present their right side. The motivation to portray emotion, or conceal it, might explain why portraits of males show a reduced leftward bias, and also why portraits of scientists from the Royal Society show no leftward bias. PMID- 10467742 TI - Restoration of ocular dominance plasticity mediated by adenosine 3',5' monophosphate in adult visual cortex. AB - Noradrenaline (NA)-stimulated beta-adrenoreceptors activate adenylate cyclase via excitatory G-proteins (Gs). Activated adenylate cyclase in turn promotes the production of cAMP. Critical roles of cAMP-dependent protein kinase A (PKA) in divergent cellular functions have been shown, including memory, learning and neural plasticity. Ocular dominance plasticity (ODP) is strongly expressed in early postnatal life and usually absent in the mature visual cortex. Here, we asked whether the activation of cAMP-dependent PKA could restore ODP to the aplastic visual cortex of adult cats. Concurrent with brief monocular deprivation, each of the following cAMP-related drugs was directly and continuously infused in the adult visual cortex: cholera toxin (a Gs-protein stimulant), forskolin (a Gs-protein-independent activator of adenylate cyclase) and dibutyryl cAMP (a cAMP analogue). We found that the ocular dominance distribution became W-shaped, the proportion of binocular cells being significantly lower than that in respective controls. We concluded that the activation of cAMP cascades rapidly restores ODP to the adult visual cortex, though moderately. The finding further extends the original hypothesis that the NA-beta-adrenoreceptors system is a neurochemical mechanism of cortical plasticity. PMID- 10467744 TI - Genomic demography: a life-history analysis of transposable element evolution. AB - Retrotransposons are ubiquitous mobile genetic elements that have played a significant role in shaping eukaryotic genome evolution. The genome of the yeast Saccharomyces cerevisiae harbours five families of retrotransposons, Ty1-Ty5. With the publication of the S. cerevisiae genome sequence, for the first time a full genomic complement of retrotransposon sequences is available. Analysis of these sequences promises to yield insight into the nature of host--transposon coevolution. Evolutionary change in Ty elements depends on their replication and excision rates, which have been determined in the laboratory. Rates measured in the laboratory may differ from those that have operated over evolutionary time. Based on an analysis of sequence data for the Ty1, Ty2 and hybrid Ty1/2 families, we develop a novel 'genomic demography' model to estimate long-term transposition and excision rates and to estimate how long ago these elements entered the yeast genome. We find that rates of excision and transposition have averaged 7.2-8.7 x 10(-8) per generation over evolutionary time. Two separate models provide upper- and lower-bound estimates for the age of the system, suggesting that the first elements entered the genome between approximately 50 million and 250 million generations ago. PMID- 10467745 TI - Microgeographical distribution of two chromosomal races of house mice in Tunisia: pattern and origin of habitat partitioning. AB - Two chromosomal races of the house mouse occur in Tunisia, a standard morph (40St) found all over the country, and a derived morph (22Rb) occurring only in central Tunisia. In this region, habitat partitioning between the two morphs was investigated by a microgeographical analysis of their distribution, assessing habitat characteristics and demographic parameters. Results showed that the 22Rb mice always occurred in the oldest sections of towns (medinas), often extending to more recent surrounding neighbourhoods where the 40St morph was most abundant. The latter was never trapped within the medinas. The transition between the two morphs was located within cities in the more recent areas, the hybrid zone being estimated at less than 0.5 km in width by a clinal analysis of chromosomal data. Although differences between habitats exist, almost no demographic differences were found between populations of the two morphs when they occurred in the same or in different habitats. Two hypotheses are discussed to account for the origin of habitat partitioning. The first relies on competitive exclusion of the 40St mice from the medinas by the derived 22Rb mice; the second is based on stochastic processes related to historical evolution of Tunisian urban communities. PMID- 10467747 TI - The fallout from Cassini. PMID- 10467746 TI - The origin of eukaryotes: the difference between prokaryotic and eukaryotic cells. AB - Eukaryotes have long been thought to have arisen by evolving a nucleus, endomembrane, and cytoskeleton. In contrast, it was recently proposed that the first complex cells, which were actually proto-eukaryotes, arose simultaneously with the acquisition of mitochondria. This so-called symbiotic association hypothesis states that eukaryotes emerged when some ancient anaerobic archaebacteria (hosts) engulfed respiring alpha-proteobacteria (symbionts), which evolved into the first energy-producing organelles. Therefore, the intracellular compartmentalization of the energy-converting metabolism that was bound originally to the plasma membrane appears to be the key innovation towards eukaryotic genome and cellular organization. The novel energy metabolism made it possible for the nucleotide synthetic apparatus of cells to be no longer limited by subsaturation with substrates and catalytic components. As a consequence, a considerable increase has occurred in the size and complexity of eukaryotic genomes, providing the genetic basis for most of the further evolutionary changes in cellular complexity. On the other hand, the active uptake of exogenous DNA, which is general in bacteria, was no longer essential in the genome organization of eukaryotes. The mitochondrion-driven scenario for the first eukaryotes explains the chimera-like composition of eukaryotic genomes as well as the metabolic and cellular organization of eukaryotes. PMID- 10467748 TI - Silicone safe. PMID- 10467749 TI - Calculating immunity. PMID- 10467750 TI - Repairing the damaged spinal cord. PMID- 10467751 TI - The throat singers of Tuva. PMID- 10467752 TI - [XVI Academic meeting of German-speaking university teachers of gynecology and obstetrics. Monastery Seeon, April 29-May 1, 1999. Abstracts]. PMID- 10467753 TI - Exploring unconventional medical systems. AB - Exotic medical systems still exist in the world's developing areas. These systems often embrace a unique pharmacopaeia and remarkable human relationships. Here is a guide for your personal exploration and appreciation of health care systems unfamiliar to Western medicine. It has been developed from personal use, offering practical suggestions for the study of unusual medical beliefs and practice in the field--and it can be used, closer to home, to survey disease and treatment concepts found in alternative or complementary medicine. PMID- 10467754 TI - Current management of stage I adenocarcinoma of the endometrium. AB - OBJECTIVE: This study was undertaken to assess the current management and outcome of patients with stage I adenocarcinoma of the endometrium. METHODS: One hundred thirty-five patients with stage I adenocarcinoma of the endometrium were treated with hysterectomy, bilateral salpingo-oophorectomy, and surgical staging. Patients were then stratified into high risk or low risk groups based on grade, depth of myometrial invasion, and the presence or absence of lymph-vascular space invasion. Postoperative treatment was then individualized based on risk assessment. RESULTS: Sixteen of 135 patients (12%) underwent postoperative adjuvant pelvic radiation. The remaining patients were treated with observation following surgery. Actuarial survival at three years was 97%. CONCLUSIONS: Surgical staging of endometria cancer provides critical information with regard to the extent of cancer and prognosis. When cancer is confined to the uterine corpus, histopathologic findings can be used to assess individual patient risk; high risk patients may then be selected for postoperative radiation. Relatively few patients will require adjuvant treatment and overall survival appears excellent. PMID- 10467755 TI - Sylvester O'Halloran Surgical Scientific Meeting. 12-13 March 1999. Abstracts. PMID- 10467756 TI - University of Limerick Biomedical Institute 2nd Annual Research Day. Biomedicine in the Aged. 25 June 1999. Abstracts. PMID- 10467757 TI - Medical sociology: a personal fifty year perspective. AB - This paper reviews the author's experience in becoming a medical sociologist before the field had become formalized. The contributions to medical sociology of sociologist Selden D. Bacon and physician and medical educator William R. Willard are described. The relationship of medical sociology to medical behavioral science, as experienced at the University of Kentucky, is discussed. Finally, the thesis of the author's 1957 paper on the nature and status of medical sociology is re-examined. PMID- 10467758 TI - Marital transitions, marital beliefs, and mental health. AB - In this paper, we examine whether people's beliefs about the permanence, desirability, and importance of marriage moderate the impact of marital transitions--including marital losses and gains--on depression, a disorder associated with both marital status and role transitions. Using two waves of panel data from the National Survey of Families and Households (N = 10,005), we find that a marital loss results in increased symptoms, whereas a marital gain results in decreased symptoms. We also find, however, that the negative effects of a marital loss are greater for people who believe in the permanence of marriage than they are for those who do not. Conversely, the positive effects of a marital gain are greater for people who believe in the desirability and importance of marriage than they are for those who do not. Our results highlight the potential utility of more systematically incorporating people's beliefs--and sociocultural factors more generally--into theory and research on the impact of stressors on mental health. PMID- 10467759 TI - Integrating sociological and biological models. PMID- 10467760 TI - Testosterone and men's depression: the role of social behavior. AB - Medical research suggests that testosterone has positive effects on mood (thereby reducing the chances of depression), and social science research finds testosterone to be related to antisocial behavior, risk behavior, unemployment and low paying jobs, and being unmarried--factors known to be positively related to depression. Analysis of a sample of 4,393 men finds a parabolic model best fits the data. The relationship between testosterone and depression is inverse for men with below average testosterone and direct for those with above average testosterone. The relationship disappears for those with above average testosterone when controls for antisocial and risk behaviors and the absence of protective factors such as marriage and steady employment are in the equation. The relationship is unchanged for those with below average testosterone. The results help explain the difference between medical and social research findings. Mechanisms accounting for the findings are explored. PMID- 10467761 TI - A biopsychosocial model of glycemic control in diabetes: stress, coping and regimen adherence. AB - This study examines stress, coping, and regimen adherence as determinants of chronic and transient metabolic control in diabetes. We also examine the interaction of biologic vulnerability and psychosocial risk factors to see if Type 1 (insulin dependent diabetes mellitus) or Type 2 (noninsulin dependent diabetes mellitus) diabetes had greater responsiveness to psychosocial risk factors. Analyses of data from insulin-treated adults with Type 1 (N = 57) and Type 2 (N = 61) diabetes supported the biopsychosocial model. For Type 1 diabetes, self-controlling persons had better glycemic control and emotional persons had worse (because of differences in stress). All of these associations were mediated by regimen compliance. For Type 2 diabetes, self-controlling persons had better glycemic control for reasons other than regimen compliance. There was an interaction between biologic and psychosocial factors, with psychosocial factors accounting for more variance in glycemic control within Type 1 patients. Stable psychosocial resources (i.e., education, being married, and positive coping styles) were associated with better chronic glycemic control, while stress and regimen nonadherence were associated with worse transient glycemic control. PMID- 10467762 TI - The conditions for state action in Florida's health-care market. AB - Despite the lack of confidence in government agencies to operate a nationalized health-care system in the United States, government agencies have significantly influenced the distribution and financing of health-care services in the market. Using the State of Florida as a case study, we examine the conditions under which a state health-care agency can consistently influence health-care market arrangements. We examined records from Florida's legislative sessions between 1965 and 1993 focusing on 27 legislative initiatives to involve the state's health-care agencies in the health-care services market. Using Boolean qualitative comparative analysis (QCA), we examined the conditions that facilitated or inhibited legislative policy initiatives for state action in Florida's health-care services market. The cohesiveness of state administrative agency and legislative leadership is of primary importance. Fragmented interests among health-care providers and fiscally legitimate policy positions, whether those of state agencies or health-care providers, are important enabling factors for state action. PMID- 10467763 TI - An institutional analysis of HIV prevention efforts by the nation's outpatient drug abuse treatment units. AB - Drawing from an institutional-theory perspective on innovations in organizations, this paper examines the use of human immunodeficiency virus (HIV) prevention practices by the nation's outpatient substance abuse treatment units during a critical period from 1988 to 1995. An institutional perspective argues that organizations adopt new practices not only for technical reasons, but also because external actors actively promote or model the use of particular practices. We examine the extent to which treatment units use several practices to prevent HIV infection among their clients and among drug-users not in treatment. Results from random-effects regression analyses of national survey data show that treatment units significantly increased their use of HIV prevention practices from 1988 to 1995. Further, the results show that treatment units' use of prevention practices was related to clients' risk for HIV infection, unit resources available to support these practices, and organizational support for the practices. Implications are discussed for an institutional view of organizational innovation as well as for research on HIV prevention. PMID- 10467764 TI - Vector competence of three Venezuelan mosquitoes (Diptera: Culicidae) for an epizootic IC strain of Venezuelan equine encephalitis virus. AB - Experimental studies were undertaken to evaluate the vector competence of selected mosquito species [Aedes taeniorhynchus (Wiedemann), Culex declarator Dyar and Knab, and Mansonia titillans (Walker)] from northwestern Venezuela for the epizootic (IC) strain of Venezuelan equine encephalitis (VEE) virus that was responsible for the 1995 outbreak of VEE in this area. Ae. taeniorhynchus was highly susceptible to infection (94% of 35), and 89% had a disseminated infection. Virus-exposed Ae. taeniorhynchus that refed on susceptible hamsters readily transmitted virus, confirming that this species was an efficient vector of VEE virus. In contrast, only 1 of 28 (4%) Cx. declarator was infected, and that individual did not develop a disseminated infection. Ma. titillans was moderately susceptible (3 of 8 infected, 38%), and 2 (25%) of these had a disseminated infection. These data indicate that Ae. taeniorhynchus was an important epizootic vector during the 1995 VEE outbreak in Columbia and Venezuela. PMID- 10467765 TI - Bite transmission of vesicular stomatitis virus (New Jersey serotype) to laboratory mice by Simulium vittatum (Diptera: Simuliidae). AB - Laboratory-reared female black flies (Simulium vittatum Zetterstedt) were infected experimentally with a 1997 vesicular stomatitis virus New Jersey serotype isolate and allowed to feed on susceptible laboratory mice. All mice exposed to black fly bite seroconverted by day 21 after infection, an indication of virus transmission. In addition, viral RNA was detected in the spleen of several mice. These findings are consistent with the hypothesis that black flies are involved in VSV-NJ transmission during epizootics in the western USA and represent the 1st confirmed example of biological transmission of an arbovirus by a member of the Simuliidae using an animal model. PMID- 10467766 TI - Comparison of methods for introducing and producing artificial infection of ixodid ticks (Acari: Ixodidae) with Ehrlichia chaffeensis. AB - Only 29.5 +/- 8.91% of engorged Amblyomma americanum (L.) nymphs that we inoculated with Ehrlichia chaffeensis molted successfully to adults compared with 75.8 +/- 7.46% of engorged nymphs that were not inoculated. However, 65.4 +/- 6.02% of unfed nymphs of this species were exposed for 2 h to E. chaffeensis suspension introduced to them through glass capillaries gained weight. These nymphs were placed on rabbits, and approximately 50% of them completed their feeding and molted successfully to adults. Weight gained was higher (71.8 +/- 17.33% and 69.8 +/- 23.26%) for unfed A. americanum females that fed from capillaries for 2 and 24, h respectively, than for nymphs. Similar values were recorded for Dermacentor variabilis (Say) (61.0 +/- 16.23%) and Rhipicephalus sanguineus (Latreille) (59.0 +/- 18.62%) females after 24 h of capillary feeding. The amount of E. chaffeensis suspension taken in by females of A. americanum, D. variabilis, and R. sanguineus during 24 h of feeding was 11.2 +/- 3.56, 10.9 +/- 4.29 and 6.3 +/- 2.35 microliters, respectively. This volume is equivalent to approximately 12,969, 12,622, and 7,295 infected cells ingested by the species mentioned above. Positive correlation between the volume taken in by the ticks and the weight gained by the females was found, but the initial weight of the unfed females did not effect the weight they gained. The pathogen was found in the females of all 3 species by polymerase chain reaction procedures for at least 7 d, indicating that the capillary feeding method can be successfully used for infecting unfed ticks. The potential use of this method is discussed. PMID- 10467767 TI - Necrophilous insect succession on rabbit carrion in sand dune habitats in northern France. AB - Six freshly killed rabbit carcasses were exposed in different habitats in the coastal dune massif of Ambleteuse (northern France) during the spring seasons of 1996 and 1997. In total, 66 arthropod species were collected during the decomposition of these carcasses, and particular attention was paid to taxa of necrophilic significance. The pattern of insect activity was recorded and is discussed in relation to meteorological conditions. One significant feature in this study was the delay in initial oviposition by flies during 1996, which demonstrates the need for caution in estimation of postmortem interval by entomological techniques in early spring. Although daily temperatures may be favorable for adult fly activity, flies may be absent because of small population size and low nocturnal temperatures during this period. PMID- 10467769 TI - Gonotrophic development in Oestrus ovis (Diptera: Oestridae). AB - Microanatomical characteristics and the size of the ovaries of Oestrus ovis L. during development were related to the intrapuparial-phenological stadia. Mature 3rd instars were collected from the head cavities of slaughtered goats, and pupae were reared under laboratory conditions. The length of freshly dissected ovaries and follicles were measured daily after pupal-adult apolysis to emergence. Ovarian tissue was stained using the PAS-Picroindigocarmine techniques. Oocyte development was classified according to a six-stage scale previously used in oestrid species. Shortly after pupal-adult apolysis, the single primary follicle is still unseparated from the germarium. In early white-eyed pharate adults, the primary follicle of stage 1 separates from the germarium, the nurse cells and oocyte are surrounded by a layer of cuboidal follicular cells, and the remaining oogonia degenerate. Oocytes in stage 2 initiate yolk deposition becoming ovoid, and this occurs when pharate adults have white-yellow to orange eyes. Oocytes in stage 3 are mainly vitellogenic, during the orange to red-eye stage. In stage 4, oocytes complete vitellogenesis and nurse cells degenerate when pupae achieve 90 96% of development. Mature oocytes of stage 5 can be found at emergence. Ovaries and ovarioles increase in size because of yolk deposition. O. ovis begins oogenesis as pharate adults, whereas vitellogenesis occurs during 55-96% of pupal development. Females emerge with one life-long complement of eggs ready to be fertilized, similar to other species of the Family Oestridae. PMID- 10467768 TI - Morphological and cytochemical characterization of female Anopheles albimanus (Diptera: Culicidae) hemocytes. AB - Hemocytes of 2- to 3-d-old female Anopheles albimanus Wiedemann are described by morphology, cytochemistry, and functional criteria. Supplemented Grace's insect medium in a modified Foley's perfusion method was used to obtain hemolymph from An. albimanus. Morphological analysis indicated 3 types of hemocytes were present, prohemocytes, plasmatocytes, and granular cells. Prohemocytes were small round cells with a high nuclear/cytoplasmic ratio. Plasmatocytes were the most abundant cell types in the hemolymph, and appeared as small to large and spindle shaped cells with round or elongate nucleus, variable number of vacuoles, small granules, and pseudopodia. Granular cells were small to large and round with a large number of cytoplasmic granules, vacuoles, and numerous filopodia. Ultrastructurally, prohemocytes were undifferentiated with abundant free ribosomes and with few small electron-dense granules. Plasmatocytes were rich in mitochondria, rough endoplasmic reticulum, free ribosomes, small electron-dense granules, numerous peripheral vacuoles and with an important organelle polarization. Granular cells contained numerous large electron-dense granular inclusions and vacuoles. Cytochemical studies showed that plasmatocytes and granular cells have cationic bactericidal proteins. Only granular cells showed phenoloxidase and probably lysosomal activities. In vitro functional studies demonstrated that both plasmatocytes and granular cells were able to attach to glass slides, and only plasmatocyte had phagocytic activity and motility. These results characterize the hemocytes of An. albimanus and suggest that plasmatocytes and granular cells may have a role in defense responses to foreign organisms. PMID- 10467770 TI - Aedes albopictus (Diptera: Culicidae): an experimental and natural host of Dirofilaria immitis (Filarioidea: Onchocercidae) in Florida, U.S.A. AB - Females of an Aedes albopictus (Skuse) colony from southeastern Florida, U.S.A., ingested low (22.9 +/- 3.2 mg/Female) and high (243.2 +/- 37.6 mf/Female) numbers of microfilariae from a dog infected with Dirofilaria immitis (Leidy). High mortality of females occurred during the first 4 d after infection regardless of the number of microfilariae ingested; daily mortality was almost negligible during 5-15 d after infection. Percentage of survival 15 d after infection was higher (63%) in females that ingested low numbers of microfilariae than those (15%) that ingested high numbers of microfilariae. The development of most of the D. immitis larvae was arrested in late L1 stage with some of the L1 stage larvae becoming melanized intracellularly in the Malpighian tubule cells of Ae. albopictus. Fifteen days after infection, development of D. immitis to the infective L3 stage occurred in only 10.9% of the surviving F1 and F2 Ae. albopictus that ingested low numbers of microfilariae, but in 94% of Anopheles quadrimaculatus Say ingesting similar numbers of microfilariae, as a control. Females of Ae. albopictus ingesting high numbers of microfilariae had more surviving females with L3 than those ingesting low numbers of microfilariae. The number of L3 larvae in the Malpighian tubules, hemocoel, head capsule and proboscis ranged from 1 to 37 per female, indicating the potential of Ae. albopictus to transmit D. immitis. Development of D. immitis larvae was not affected by co-infection with Ascogregarina taiwanensis (Lien & Levine), although both parasites infect the Malpighian tubules, the first intracellularly and the second extracellularly. After one generation of selection, a strain of Ae. albopictus susceptible to D. immitis developed 2.5 times more L3 than the parent strain. These results show that a small portion of the natural population of Ae. albopictus is susceptible to infection with D. immitis and that susceptibility may be increased rapidly by selection. The presence of developing Dirofilaria sp. larvae in the Malpighian tubules of field-caught females indicated that Ae. albopictus may be infected naturally with D. immitis in Florida. PMID- 10467771 TI - Occurrence of Orientia tsutsugamushi in chiggers (Acari: Trombiculidae) and small animals in an orchard near Bangkok, Thailand. AB - The ecology of Orientia tsutsugamushi (Hayashi) was studied in chiggers and small animals in an orchard near Bangkok, Thailand. Small animals were trapped monthly between July and November 1992 and examined for the presence of O. tsutsugamushi and ectoparasitic chiggers. A total of 40 Rattus rattus (L.) and 16 Tupaia glis (Diard) was trapped. O. tsutsugamushi was isolated from liver and spleen samples of 30.8% of R. rattus and 18.6% of T. glis. Antibodies to O. tsutsugamushi were detected in 95% of R rattus, and IgG antibodies persisted for up to 10 mo after removing rats from potential reinfection in the field. A total of 1,133 chiggers was identified and examined for the presence of O. tsutsugamushi using a direct fluorescent antibody test, and 2.6% Leptotrombidium deliense Walch, 5.1% Aschoshoengastia indica Hirst, 2.6% A. (Laurentella) sp. #2 and 0.9% A. (Laurentella) sp. #4 were infected. Forty-four pools of chiggers from these animals were triturated and injected into mice. Seven pools were obtained from T. glis and 1 was positive for O. tsutsugamushi, and 3 of 37 pools from R. rattus were positive. A proportion of the engorged chiggers collected was reared to the adult stage, and the progeny of these adults tested for the presence of O. tsutsugamushi. The progeny of 186 females was tested, and the progeny of 2 L. deliense was found to be naturally infected with O. tsutsugamushi. PMID- 10467772 TI - Scanning electron microscopy of human head louse (Anoplura: Pediculidae) egg and its clinical ramifications. AB - Pediculosis affects more elementary school students than all other communicable childhood diseases combined, excluding the common cold. The current study using scanning electron microscopy visualized operculums through which developing embryos received oxygen, as well as fracture lines within the adherent sheath by which the egg is attached to the hair. Using microscopic techniques, including freeze-fracturing, the mode of attachment of the sheath to the hair follicle was observed, in addition to the existence of several inner membranes enclosing the embryo within the egg structure. The chemical nature of the sheath is also analyzed using electron dispersive x-ray analysis. PMID- 10467773 TI - Fluctuating hydrating and dehydrating relative humidities effects on the life cycle of Dermatophagoides farinae (Acari: Pyroglyphidae). AB - Reducing relative humidity to < 50% in homes is recommended as one means of reducing dust mite populations in the homes of those who suffer allergies to house dust mites. Because of some activities in the home (e.g., bathing, cooking, opening windows), it may not be possible to keep relative humidity constantly < 50%. We determined how the fluctuating daily regimes of hydrating (75%) and dehydrating (35%) relative humidities affected the development of Dermatophagoides farinae Hughes. The life cycle was completed (egg to adults) when mites were given regimes of 24 h at 75% RH, 8 h at 75% and 16 h at 35% RH, 6 h at 75% and 18 h at 35% RH, and 4 h at 75% and 20 h at 35% RH. The time required to complete development was inversely related to the amount of moist air given daily. Development took 58.3 +/- 1.44, 64.7 +/- 1.87, and 82.4 +/- 2.39 d for 8, 6, and 4 h of moist air daily, respectively. In comparison, the life cycle was completed in 41.1 +/- 0.50 d when development occurred at a constant 75% RH. Egg incubation time was significantly longer for fluctuating ambient relative humidity compared to a continuous 75 or 35% RH. Of the emerging larvae 53.8, 72.7, 62.7, and 85.0% completed the life cycle when given 4, 6, 8, and 24 h 75% RH daily and 35% RH for the remainder of the day. This study revealed that D. farinae can complete development when given only short periods of moist air daily but the rate of development is much slower than development at a constant 75% RH. Therefore, reducing ambient relative humidity does reduce the rate of development of mite populations and the accumulation of dust mite allergen. PMID- 10467774 TI - Sugar meal composition of five north central Florida mosquito species (Diptera: Culicidae) as determined by gas chromatography. AB - Gas chromatography was used to analyze sugars found in individual crops fed on by wild caught adult Anopheles quadrimaculatus s.l. (Say), Coquillettidia perturbans (Walker), Culex nigripalpus Theobald, Culiseta melanura (Coquillett), and Psorophora ferox (von Humboldt) from several north central Florida locations. A wide range of sugars was found, including fructose, glucose, sucrose, maltose, turanose, melezitose, trehalose, raffinose, erlose, and traces of arabinose, rhamnose, and several unknowns. The frequency of individuals testing positive for crop sugars ranged from 9-10% in An. quadrimaculatus and Ps. ferox to 46% in Cq. perturbans. Based on the presence of melezitose or erlose in the crop, honeydew feeding was shown to be an important dietary component in An. quadrimaculatus (57%), Cs. melanura (31%), Cx. nigripalpus (15%), Cq. perturbans (10%), and Ps. ferox (7%). Gas chromatography of crude crop contents is a rapid one-step process that can address important questions such as the source and occurrence of sugar feeding in Diptera. Unique sugar feeding field observations were recorded for several mosquito species. PMID- 10467776 TI - Culex (Acallyntrum) miyagii (Diptera: Culicidae): new species from Seram Island, Indonesia, with keys to the species of the subgenus. AB - Culex (Acallyntrum) miyagii is described as a new species from Seram Island, Indonesia. The adult male, female, pupa, and larva are described in detail, and illustrations of the male genitalia, pupa, and larva are provided. Cx. miyagii is a forest species breeding in inflorescences of Costus sp. (Costaceae). Keys are provided for identifying the females and 4th-instar larvae of the species of Acallyntrum. PMID- 10467775 TI - Mosquito fauna in water-holding containers with emphasis on dengue vectors (Diptera: Culicidae) in Chungho, Taipei County, Taiwan. AB - A survey was conducted to characterize the mosquito fauna in water-holding containers in residential and open areas with emphasis on Aedes albopictus Skuse, and to determine the prevalence of mosquito-positive containers to obtain background information for a source reduction program. Seven container-inhabiting mosquito species were collected in residential areas, including Ae. albopictus, Culex quinquefasciatus Say, C. pallidothorax Theobald, C. neomimulus Lien, C. bicornutus Theobald, C. fuscanus Wiedemann, and Tripteroides aranoides Theobald. Ae. albopictus was the most abundant species and was collected in 18.49% of the containers examined. Other mosquito species were found in 0.02-0.16% of the total numbers of water containers checked. Of the positive water containers, 97.33% contained Ae. albopictus larvae. Sixteen species were collected in open areas away from residential sites. Ae. albopictus was most abundant and was present in 24.50% of the total number of containers examined. C. bicornutus, C. quinquefasciatus, and C. pallidothorax were present in 2.17, 1.48, and 1.08%, respectively. Ae. albopictus larvae were found in 74.78% of the mosquito-positive water containers. The mean Breteau index (+/- SE) in 1997 (17.22 +/- 1.59 mosquito-positive containers per 100 premises) per village in residential areas was not significantly higher than the index in 1996 (16.86 +/- 1.33). The frequency of occurrence of Ae. albopictus in different containers is presented. A greater preference for containers in outdoor habitats was found. The ratio of the total number of containers indoors and outdoors per premises was 1.15:1.00, whereas the ratio of mosquito-positive containers indoors and outdoors was 1.00:1.83. Most indoor breeding containers (86.11%) in residential areas were used for planting purposes and for recreation (12.50%). The number of breeding containers in residential areas that were classified as trash increased from 0.35% indoor to 39.03% outdoor, whereas the number used for planting reduced to 34.95%. Most breeding containers (63.02%) in open areas were trash; natural containers (11.86%) were the 2nd most common breeding site followed by temporarily unused (8.84%), watering (6.98%), and planting containers (5.81%). PMID- 10467777 TI - Ectoparasites of Brazilian free-tailed bats with emphasis on anatomical site preferences for Chiroptonyssus robustipes (Acari: Macronyssidae). AB - Seven species of arthropods were recovered from 45 Brazilian free-tailed bats, Tadarida brasiliensis (I. Geoffrey), captured from May 1994 to May 1995 from a roost in Jenkins County, GA. Six species of mites were found infesting the bats, representing 5 families: Macronyssidae, Cheyletidae, Sarcoptidae, Rosensteiniidae, and Uropodidae. The macronyssid mite chiroptonyssus robustipes (Ewing) and the hemipteran bat bug, Cimex adjunctus (Barber), were collected from the host and the roost. C. robustipes was the most abundant mite (2,508 specimens) and the macronyssid Steatonyssus ceratognathus (Ewing) (189 specimens) was the 2nd most abundant mite found infesting the bats. The predatory cheyletid mite Cheyletonella vespertilionis Womersley (51 specimens) was also collected from the bats, and this is thought to be the 1st report of this mite from Georgia. In addition, Teinocoptes sp. (Sarcoptidae, 8 specimens), Nycteriglyphus sp. (Rosensteiniidae, 8 specimens), and uropodid mites (5 specimens) were removed from the bats. Of the 45 bats examined, all 30 females and 14 of 15 males were infested with C. robustipes. Twenty-three bat anatomical areas were designated and all C. robustipes mites were counted to ascertain anatomical preference. This mite was significantly more abundant on the ventral wing membrane areas posterior to the radiusulna and on the lower torso. Protonymphs of C. robustipes predominated on the wing membranes of the bats, whereas adult mites were more concentrated on the torso and head. PMID- 10467778 TI - Mode of action of lufenuron in adult Ctenocephalides felis (Siphonaptera: Pulicidae). AB - When cat fleas, Ctenocephalides felis (Bouche), were fed concentrations of lufenuron in cattle blood ranging from 0.5 to 4 ppm, adult mortality increased in a dose-dependent manner to a maximum of approximately 24% over a period of 10 d. Fleas treated with 0.5 ppm produced abnormal endocuticle consisting of protein globules embedded in an amorphous chitin matrix. At concentrations of 1.0 ppm or greater, endocuticle formation was inhibited. Ultrastructural studies demonstrated that inhibition of chitin synthesis was associated with degeneration of the epidermal cells. The amount of epidermal cytoplasm decreased and cytoplasmic organelles including mitochondria, ribosomes, and golgi showed lytic changes. At least some mortality of treated fleas was likely the result of a weakened endocuticle and the corresponding decrease in resiliency of the cuticle to expansion during blood-feeding and egg production. An unexpected result of lufenuron treatment was the inhibition of midgut epithelial cell differentiation. At concentrations of 0.5 and 1.0 ppm, partially differentiated epithelial cells were seen in the midgut of bloodfed fleas along with fully differentiated cells. PMID- 10467779 TI - Sensilla of the antennae and maxillary palps of Culicoides hollensis and C. melleus (Diptera: Ceratopogonidae). AB - The types and arrangement of the sensilla on the female antennae and maxillary palps of both sexes of 2 estuarine species of biting midges, Culicoides hollensis (Melander & Brues) and C. melleus (Coquillett), were studied by scanning electron microscopy. Five sensillum types are found on the antennae of both species. Both species have sharp-tipped sensilla trichodea and sensilla basiconica only on subsegments 9-13 and blunt-tipped sensilla trichodea on all subsegments. Sensilla coeloconica occur on subsegment 1 of both species and various other subsegments depending on the species. Sensilla chaetica are found on all subsegments of C. melleus, but only on subsegments 1-8 and 13 on C. hollensis. Both sexes of C. hollensis have either 1 or 2 pits on the 3rd segment of the maxillary palps, which contain 8-15 bulb-shaped sensilla in females compared with 5-6 in males. No pit is found on the maxillary palps of either sex of C. melleus, but rather the bulb-shaped sensilla are distributed individually on this segment, ranging from 13 to 29 on the female and 6-10 on the male palps. These data are compared with data obtained from previously published light and electron microscopical studies of biting midge sensory structures. PMID- 10467780 TI - Attraction of Culex pipiens pipiens (Diptera: Culicidae) to flower volatiles. AB - Methylene chloride extracts of flowers from 4 plants (common milkweed, Asclepias syriaca L; ox-eye daisy, Leucanthemum vulgare Lamarck; Canada goldenrod, Solidago canadensis L.; and yarrow, Achillea millefolium L.) were tested in a dual-port olfactometer for attractiveness to female northern house mosquitoes, Culex pipiens pipiens L. Fifty milligrams of extract was used for each test, which was equivalent to the amount of extract from flowers of 0.58 common milkweed inflorescences, 1.75 ox-eye daisy flower heads, 0.33 Canada goldenrod plants, or 3.14 yarrow plants. Significantly more Cx. pipiens responded to the common milkweed flower extract than to a control. Ox-eye daisy, Canada goldenrod and yarrow flower extracts were not significantly attractive to this species. A headspace sample of the common milkweed extract was collected by solid phase microextraction and analyzed by gas chromatography-mass spectrometry. The most abundant compounds in the headspace sample were 2-phenylethanol and benzyl alcohol. Analysis of the common milkweed extract and standards of 2-phenylethanol and benzyl alcohol by gas chromatography-flame ionization detection confirmed that these compounds were present in the extract. Cx. pipiens did not respond to synthetic blends of these 2 compounds when tested in a dual-port olfactometer. PMID- 10467781 TI - Vector competence of Aedes notoscriptus (Diptera: Culicidae) for Barmah Forest virus and of this species and Aedes aegypti (Diptera: Culicidae) for dengue 1-4 viruses in Queensland, Australia. AB - Aedes notoscriptus (Skuse) mosquitoes colonized from Brisbane, Queensland, Australia, were fed on blood suspensions containing either Barmah Forest (BF) virus, dengue virus type 1 (DEN-1), dengue virus type 2 (DEN-2), dengue virus type 3 (DEN-3), or dengue virus type 4 (DEN-4). Ae. notoscriptus was shown to be moderately susceptible to oral infection with BF virus, ID50 = 10(3.3) CCID50 per mosquito, with titers in infected mosquitoes peaking 10 d after infection. BF virus transmission occurred between days 5 and 12, with an average rate of 45% (range, 40-50%). For DEN 1-4, Ae. notoscriptus showed low level susceptibility to oral infection, with maximum head and thorax squash infection rates of 8, 5, 13, and 3%, respectively. Rates of infection were notably higher in a north Queensland strain of Aedes aegypti (L.) used as a positive control, with maximum thorax squash infection rates of 58, 68, 78, and 60% for DEN 1-4, respectively. On the basis of these data, it is concluded that Ae. notoscriptus could act as an efficient vector of BF in urban environments, but it is unlikely to be an important vector of DEN. PMID- 10467782 TI - Spatial and temporal analysis of Ross River virus disease patterns at Maroochy Shire, Australia: association between human morbidity and mosquito (Diptera: Culicidae) abundance. AB - Notifications of Ross River (RR) virus disease in Maroochy Shire were mapped according to the patient's place of residence, and standardized morbidity ratios were calculated for each of 11 census districts, with 4 areas having higher than average overall rates of RR virus disease notification. Temporal analysis of RR virus disease notifications from each of the 11 areas indicated that epidemics of RR virus disease either were widespread, resulting in higher than average numbers of cases from the majority of areas, or limited to only certain areas. Mosquitoes were collected in CO2 light traps, and the numbers analyzed to determine whether there was any association between mosquito abundance and disease incidence. Seasonal light trap indices for the fresh water-breeding Culex annulirostris Skuse and brackish water-breeding Aedes funereus (Theobald) were correlated positively with disease incidence. There was no apparent association between Aedes vigilax (Skuse) or Aedes procax (Skuse) light trap indices and disease incidence. Identification of human subpopulations with higher than average rates of RR virus disease, and the mosquito vectors associated with virus transmission, should provide a framework for the development of focused and therefore more effective control programs. PMID- 10467783 TI - Mosquito (Diptera: Culicidae) host compatibility and vector competency for the human myositic parasite Trachipleistophora hominis (Phylum Microspora). AB - Microsporidian spores of Trachipleistophora hominis Hollister, isolated from a human, readily infected larval stages of both Anopheles quadrimaculatus Say sensu lato and Culex quinque-fasciatus Say. Mosquito infections with T. hominis were located, primarily, in abdominal muscles in segment numbers 4 through 6; other spores were found in the hemocoel and proboscis. Nearly 50% of the infected mosquito larvae survived to the adult stage. Spores recovered from adult mosquitoes were inoculated into mice and resulted in significant muscle infection at the site of injection. Preliminary observations also showed that T. hominis spores can be passively transferred from infected mosquitoes to a sugar water substrate. PMID- 10467784 TI - Squalene: a naturally abundant mammalian skin secretion and long distance tick attractant (Acari: Ixodidae). AB - Squalene is a naturally occurring lipid on mammalian skin and is an attractant to the lone star tick, Amblyomma americanum (L.). In this study, we compared squalene to the standard tick-attractants, benzaldehyde, isobutyric acid, methyl salicylate, nonanoic acid, and o-nitrophenol identified as active ingredients of tick aggregation-attachment pheromones and determined its effectiveness in field and laboratory settings at varying distances. Squalene was detected from 1/4 m greater than the standard tick attractants, attracted a greater percentage of ticks (75 compared with 0-43%) and featured a rapid response time (< 30 min). Thus, squalene contributes more to the tick's ability to locate hosts at greater distances than aggregation-attachment pheromones. These results have important implications for improving tick monitoring and control programs by adding squalene as a supplement to existing attractant baits. PMID- 10467785 TI - Histochemical analysis of the nit of Pediculus humanus capitis (Anoplura: Pediculidae). AB - Histochemical stainings of head louse nits were performed and observations indicated that the nit sheath was nonchitinous. One discernable membrane composed of hyaluronic acid was established that encircled the larvae within the egg. The structural framework of the egg included a definite lattice around each aeropyle of the operculum, through which oxygen exchange occurred. PMID- 10467786 TI - Maternal mortality in the developing world: many unanswered questions. PMID- 10467787 TI - Rates of endometriosis recurrence and pregnancy 1 year after treatment with intranasal buserelin acetate (Suprecur) (a prospective study). AB - OBJECTIVES: This multicenter prospective study of buserelin acetate (Suprecur) therapy for endometriosis was performed in order to examine pregnancy rates and the recurrence of endometriosis 1 year after the end of intranasal administration. METHODS: This study was conducted at 349 institutions, including university hospitals and major hospitals in Japan, and 1,284 patients were recruited as participants. Recurrence of symptoms was followed for 1 year after the end of administration of buserelin acetate in patients whose condition as a whole was evaluated as "improved" or better at the end of administration. Furthermore, the pregnancy rate was similarly followed in patients who complained of sterility at the commencement of the administration. Recurrence was defined as either a case in which all of the main symptoms observed before administration returned to the pretreatment state, or a case in which at least one symptom deteriorated compared to the pretreatment state. Kaplan-Meier's method was used for statistical analysis. RESULTS: Among 459 patients who could be evaluated for recurrence of symptoms, the cumulative recurrence rate was 17.8% 1 year after the end of administration. Among 156 patients who could be evaluated for pregnancy, the pregnancy rate was 23.3% 1 year after the end of administration. CONCLUSION: Intranasal buserelin acetate therapy exhibited not only transient clinical effects during administration or immediately after the end of administration, but also exhibited specific continuing effects after the end of administration. PMID- 10467788 TI - Survey of physical abuse during pregnancy GMCH, Nagpur, India. AB - INTRODUCTION: Domestic violence is a major health concern for women globally. Violence may commence or escalate during pregnancy and may be more frequent than obstetric complications e.g. preeclampsia or gestational diabetes for which they are routinely screened. No literature is available on abuse during pregnancy from India. OBJECTIVE: To assess the frequency and severity of physical abuse during pregnancy and its association with entry to antenatal care and to identify lifetime risk factors. STUDY DESIGN: Cross sectional study. STUDY SETTING: Secondary and tertiary care public hospital. PARTICIPANTS: Six hundred randomly selected pregnant women in third trimester of pregnancy (28-40 weeks of gestation) attending routine antenatal clinic were assessed for physical abuse using Abuse Assessment Screen (AAS) developed by McFarlane. MAIN OUTCOME MEASURES: Abuse status, gestational age at first antenatal visit and lifetime risk factors. ANALYSIS: Descriptive, Univariate and MLR. RESULTS: Of the 600 pregnant women interviewed, 152 (25.33%) women reported abuse during non pregnant state, 132 (22%) women reported physical abuse during index pregnancy and 10 (8.33%) women reported increase in abuse during pregnancy. In majority the abuse site was head and neck and 20% reported being hit in the abdomen. Abuse was recurrent in 92% of women. Six (4.54%) women were hospitalized, 5 (3.78%) needed medical assistance and 19.7% women were afraid of partner (husband). Abused women were twice as likely to begin antenatal care after 32 weeks of gestation as compared to non abused women (OR = 2.5; 95% CI 1.1-5.6, p = 0.02). Abused women were more likely to be Buddhist (p = 0.005), of greater parity with unwanted pregnancy, dwelling in slums (p = 0.005), from extended family (p = 0.005), with low level of education to husband (p = 0.009) and with stress to husband (p = 0.03). CONCLUSIONS: There is significant physical abuse during pregnancy in central India. Routine screen by AAS and a cohort study looking at adverse outcome of pregnancy is recommended. PMID- 10467789 TI - Serum vitamin A and E in pregnant women with hemoglobinopathies. AB - OBJECTIVE: To evaluate the relationship between the status of serum vitamin A, E and hemoglobinopathies among Thai pregnant women. METHODS: This was a cross sectional study in which serum vitamin A and E were assessed in 323 pregnant women with normal hemoglobin and 73 with hemoglobinopathies (47 with hemoglobin E and 26 with thalassemia) during the first trimester. RESULTS: There were no significant differences in the mean serum vitamin A, E concentrations and vitamin E/cholesterol ratio between pregnant women with normal hemoglobin and hemoglobinopathies, while confounding variables that might affect serum vitamin levels i.e. maternal age, gravida, BMI, gestational age, hematocrit, hemoglobin, mean corpuscular hemoglobin concentration and blood group were not different. CONCLUSION: The results of this study suggest that antenatal care in terms of micronutrients-vitamin A, E in Thai pregnant women with hemoglobinopathies should not be different from normal pregnant women. PMID- 10467790 TI - A prospective study on the effects of reformulated 2-rod Norplant implant on haemostasis after five years of use. AB - OBJECTIVE: To evaluate the long-term effects of the new reformulated 2-rod Norplant implant on haemostasis in a prospective group of subjects who have completed 5 years of use. METHODS: Data from 11 women who have completed 5 years' use of the new reformulated 2-rod subdermal implant from the original 16 women who were recruited and randomized to receive this new improved implant in a comparative study were analysed. Clinical assessment and serial blood sampling were done prior to insertion of implant and after 1, 3, 12, 18, 24, 36, 48 and 60 months of implant use. Each subject served as its own control and Analysis of Variance with Student-Newman-Kuels test was used for statistical analysis. The following parameters were determined: plasminogen activators (t-PA, u-PA), plasminogen activator inhibitor-1 (PAI-1), D-dimer, beta-thromboglobulin (beta TG), thrombin-antithrombin (TAT)-complex, fibrinogen, Factor VII, platelets, haematocrit and haemoglobin levels. RESULTS: No significant change was observed for t-PA levels in prolonged implant use. u-PA antigen showed a significant decrease whilst D-dimer were significantly elevated at only 24 months of implant use compared to pre-implant level. PAI-1 levels were not significantly changed but fibrinogen and FVII levels increased at 36 months and 42 months of use with enhanced platelet activation shown by beta-TG levels at 24 months. Platelet numbers were not affected by prolonged implant use. Haemoglobin concentration and haematocrit level showed significant fluctuations and then return to pre-implant level by 54 and 60 months. CONCLUSION: Enhanced fibrinolysis with platelet activation at 24 months of implant use were seen during the 60 months of 2-rod reformulated Norplant implant use. Hypercoagulable state was not observed although fibrinogen and FVII levels remain above the pre-implant levels as coagulation activation was not enhanced. The increased haemoglobin and haematocrit levels seen indicate enhanced bone marrow activity. There was no association between the use of reformulated 2-rod Norplant implant over 60 months of use and prothrombotic state. PMID- 10467791 TI - Early discharge: its impact on low-income mothers and newborns in the state of Hawaii. AB - OBJECTIVES: The objectives of this survey were to investigate whether, in the state of Hawaii, early discharge had a negative effect on the health of new mothers and their infants, especially those at high sociodemographic risk, and whether early discharge was implemented in compliance with the early-discharge guidelines of the American Academy of Pediatrics (AAP), and the American College of Obstetricians and Gynecologists (ACOG). METHODS: This statewide survey was conducted from June 17 through July 12, 1996. The data was collected and recorded by perinatal providers at 14 PSS (Perinatal Support Services) sites through interviews of mothers using questionnaires. This report is based on the 108 questionnaires that were returned from the PSS sites. A descriptive analysis was performed, using the statistical/epidemiologic program EPI INFO 6. RESULTS: The survey reveals that the practice of early discharge had been implemented extensively but not in compliance with AAP/ACOG early discharge guidelines. The neonatal readmission rate for this survey group was 6.5% among low-income mothers and infants. This was found to be among the highest rates reported in the United States and Canada; in the United States, reported neonatal readmission rates vary from 1 to 4%. CONCLUSIONS: Early discharge greatly contributes to negative outcomes among low-income mothers and infants. This paper reports on the results of this survey in Hawaii and recommends a comprehensive approach for promoting the health and well-being of low-income mothers and infants. PMID- 10467792 TI - Emergency peripartum hysterectomy at the Princess Badeea Teaching Hospital in north Jordan. AB - OBJECTIVE: To study the prevalence, indications and outcome of emergency peripartum hysterectomy in women delivered at the Princess Badeea Teaching Hospital in North Jordan. METHOD: This is a retrospective study of all cases of emergency peripartum hysterectomy performed between 1st of January 1994 and 31 August 1998. RESULTS: During the study period there were a total of 21 emergency peripartum hysterectomy were performed. The overall incidence was 0.5/1,000 deliveries. The mean age of patients was 34.7 +/- 3.9 years, the median parity was 6 and the mean gestational age was 36.9 +/- 2.01 weeks. There were 19 cases of caesarean hysterectomy. The leading indication for caesarean section was previous caesarean section (89.5%), placenta previa alone (10.5%). It should be noted that 7 cases with previous caesarean section also had placenta previa (41.2%). The main indications for emergency hysterectomy were, abnormally adherent placenta was the leading indication (38.1%), followed by rupture uterus (33.3%), haemorrhage and uterine atony occurred in 14.3% of cases each, maternal complications occurred in 42.9% of cases postoperatively. There were 4 cases of stillbirths and 2 cases of neonatal deaths. CONCLUSION: Peripartum hysterectomy remains a necessary procedure for life saving during abdominal and vaginal deliveries. The procedure itself is usually associated considerable perioperative morbidity. Obstetricians should identify patients at risk and anticipate the procedure and complications. PMID- 10467793 TI - Choriocarcinoma co-existent with an intact pregnancy: case report and review of the literature. AB - A patient presenting with antepartum haemorrhage due to a vaginal metastasis of choriocarcinoma is reported. Following delivery she was successfully treated with chemotherapy and surgery. This report details this unusual presentation and reviews the literature on this rare condition. PMID- 10467794 TI - Sonographically guided intralesional antibiotic injection for treatment of a recalcitrant pelvic abscess: a case report. AB - A pelvic abscess occurred in an infertile woman with an endocervical gonococcal infection after hysterosalpingographical examination. The pelvic abscess was not cured despite rigorous antimicrobial chemotherapy until two intra-abscess ceftriaxone injections were administered. This shows that antibiotics administered systemically may not reach therapeutic concentration in an abscess and an intra-abscess antibiotic injection may help to cure it. PMID- 10467795 TI - Optimal dose of nalbuphine for treatment of intrathecal-morphine induced pruritus after caesarean section. AB - OBJECTIVE: To search for the optimal dosage of nalbuphine relief of intrathecal morphine induced pruritus after caesarean section. METHODS: Ninety parturients who developed moderate to severe pruritus caused by intrathecal morphine after caesarean section were randomly allocated into 3 groups receiving 2, 3 and 4 mg of intravenous nalbuphine respectively. The improvement of pruritus and adverse effects of nalbuphine such as increasing pain scores, nausea, vomiting, sedation and respiratory depression were evaluated at 15 minutes after nalbuphine administration. RESULTS: Percentage of successful treatment of pruritus with 2, 3 and 4 mg, nalbuphine were 86.7, 96.7 and 100, respectively (p = 0.12). There was no statistically significant difference in adverse effects. No evidence of respiratory depression was detected. However, there was significant increase in pain scores in group of 4 mg nalbuphine (p = 0.004). CONCLUSIONS: Nalbuphine of 2 to 3 mg was considered to be adequate in treatment of intrathecal morphine induced pruritus after caesarean section without increasing pain scores or causing other side effects. PMID- 10467796 TI - Sexual transmission and reinfection of group B streptococci between spouses. AB - OBJECTIVE: To study how GBS infection takes place between pregnant GBS-carriers and their husbands. METHODS: Pregnant women in whom GBS infections could be detected during 26 to 30 weeks of pregnancy and their husbands were studied during the two periods of August 1994 through May 1995 (Period A, 243 couples) and June through September 1997 (Period B, 141 couples). A urine sample was collected from a husband in the same morning when the vagina of his wife was tested for GBS. GBS were also classified according to their serotypes in 34 couples during Period A and B. RESULTS: In the two periods, GBS was detected in 18.1 and 19.3% of the wive's vaginal cultures, and in 19.1 and 17.0% of husbands' urinary cultures, respectively. There were no significant differences of the rate of GBS detection between the spouses, and also between the two trials. A high possibility of GBS infection was found in a couple when either of the spouses was possible to GBS. The serotypes of 31 of the 34 couples (91.2%) were identical. CONCLUSION: It is suggested that GBS can be sexually transmitted, and cause reinfection between spouses in spite of antepartum medication. PMID- 10467797 TI - Study on genital tract Chlamydia trachomatis and gonococcal infection in Han and minority (Naxi and Dai) women in China's two provinces. AB - OBJECTIVES: To estimate the prevalence of lower female genital tract chlamydial (GTC) and gonococcal infection in 3 different ethnic groups in China, and to analyze the risk factors influencing these infections. METHODS: A cross-sectional survey + physical and laboratory examinations. 1,800 married women aged 18-35 Han (600, from Dujiangyan city, Sichuan), Naxi (600, from Yanyuan county, Sichuan), and Dai (600, from Jinghong Prefecture, Yunnan) were investigated between March 1996 to March 1997. Information was obtained about their personal history, socioeconomic status, medical history, STDs, sexual history, contraceptive history, results of both examinations. Tabulation, t-test, Chi-square test, multiple-logistic models were used to analyze the data. RESULTS: 3.2% of Han, 12.8% of Naxi and 4.5% of Dai women tested positive for gonococci; and 9.1% of Han, 16.7% of Naxi and 27.5% of Dai women tested positive for GTC infection. The differences of GTC and gonococcal infections between Han and Naxi, and Han and Dai women were significant. CONCLUSIONS: STDs are common in China today; both GTC and gonoccocal infections are related to non-use of condom, having sexual partners other than husband, number of abortions, and low education; and condom has a function in prevention of STDs. PMID- 10467798 TI - Joining the efforts to prevent and curb youth violence. PMID- 10467799 TI - Editor's choice: the plight of a people dispossessed: caring for street children in the mountains of Bulgaria. AB - This article was born out of a reflective diary kept by the author while working on a cross-community project in Northern Ireland. While it maps personal experiences, little did the author know its value in gaining insight and understanding of the eastern European situation that has now erupted with such devastating consequences. PMID- 10467800 TI - Nursing's role in NetWellness: a children and adolescent health information resource via the Web. AB - Nurses are in the business of health information. A nursing entrepreneurial model of service for children and adolescents is providing health education and health promotion through the World Wide Web. Developed at the University of Cincinnati, and later joined by The Ohio State University and Case Western Reserve University, NetWellness (http:/(/)www.netwellness.org) offers easily accessible and reliable health care information. The Ask an Expert feature is valued by parents, families, children and health professionals around the United States and the world. As members of the NetWellness expert team in Ohio, nurses are leaders in this initiative. PMID- 10467801 TI - The experiences of Hong Kong Chinese parents of children with acute lymphocytic leukemia. AB - The purpose of this study was to explore the experiences of Chinese parents of children diagnosed with acute lymphocytic leukemia (ALL). Respondents consisted of a convenience sample of nine mothers and eight fathers. Data were collected through in-depth interviews and were scheduled to coincide with the disease trajectory, and were analyzed using the matrix system described by Miles and Huberman. Major categories identified include parental reactions, methods used by the parents to disclose the child's diagnosis to others, changes in the family routine, and the preferred sources of parental support. Implications for health professionals include a need for thorough psychosocial assessment of the affected children, parents, and siblings. Limitations of the study and recommendations for future research are also discussed. PMID- 10467802 TI - Hispanic mothers' knowledge and care of their children with respiratory illnesses: a pilot study. AB - This pilot study was designed to identify Hispanic mothers' knowledge and health practices regarding caring for their children with respiratory illnesses. Using an interview guide, the researchers identified the mothers' perceptions of their children's health, the etiology and symptoms of asthma and other respiratory illnesses, health practices, folk illnesses, services sought and language barriers when using health services. The findings suggest that Hispanic mothers have limited knowledge of respiratory illnesses and use culturally related practices when caring for their sick children. Implications for pediatric nursing practice are discussed, and strategies to improve care for Hispanic families are presented. PMID- 10467803 TI - Fathers' scores on the Nursing Child Assessment Teaching Scale: are they different from those of mothers? AB - Fathers of 103 Canadian infants (ages 2 to 12 months) were observed interacting with their child using the Nursing Child Assessment Teaching Scale (NCATS). Compared with the NCATS reference data for 387 mothers of similar-aged infants with similar marital status and ethnic background, there was no significant difference on the overall total score. However, fathers had significantly lower scores than mothers on the total parent score. Infants interacting with the fathers had significantly higher total child scores than those interacting with the mothers. Although mothers and fathers were equally sensitive to their infant's cues, fathers were less contingent in interaction with their infants. Implications for practitioners are discussed. PMID- 10467804 TI - Assessing culture: pediatric nurses' beliefs and self-reported practices. AB - Assessment of culture is an essential step in providing effective nursing care to a culturally diverse patient population. The purpose of this study was to describe the beliefs and self-reported practices of pediatric nurses from four Children's Hospitals regarding the process of assessing culture. The 584 respondents indicated they worked with a culturally diverse patient population and frequently experienced cultural conflicts. Findings indicated cultural assessments were not routinely performed. The majority indicated that learning more about the child's culture would improve the effectiveness of their nursing care. Respondents were willing to conduct cultural assessments on selected patients if this task could be accomplished in less than 15 minutes. Recommendations include guidelines for developing and implementing an appropriate cultural assessment process within a hospital setting. PMID- 10467805 TI - The Healthy Start Program: mobilizing to reduce infant mortality and morbidity. PMID- 10467806 TI - A conference for the education of families who have a member with phenylketonuria. PMID- 10467807 TI - Use of the heart rate monitor to modulate physical activity in adolescents with congenital aortic stenosis: an innovative approach. AB - The task of setting exercise limitations on children with aortic stenosis is fraught with difficulties. In particular, teenagers are difficult to manage because of the increasingly professional demands of adolescents sports; the rapid changes in somatic growth, which are often accompanied by an increase in the severity of aortic valve disease; and the natural tendency of teenagers to disregard advice from authoritarian sources like a medical team. This article describes our innovative approach of using a heart-rate monitor as a means of modulating physical activity in adolescents with mild to moderate aortic stenosis. This approach enabled the setting of clear, precise, observable, measureable limits on physical activity, and self-controling of an acceptable level of physical activity. This created a different negotiation between the patient, his parents, and the medical team, and eased the concern and anxiety of the mothers. PMID- 10467808 TI - The new "Mickey Finn specials"--should GHB and ketamine be banned? PMID- 10467809 TI - The Lazarus syndrome: a second chance for life with HIV infection. PMID- 10467810 TI - APhA drug treatment protocol: management of pediatric convulsive status epilepticus. PMID- 10467811 TI - Pharmacist counseling on nutrition and physical activity--Part 1 of 2: Understanding current guidelines. PMID- 10467812 TI - Asthma patients' satisfaction with the frequency and content of pharmacist counseling. AB - OBJECTIVE: To assess the level of expectation and satisfaction of patients with asthma regarding the counseling provided by community pharmacists, and to identify the counseling information patients consider important in helping them manage their asthma. DESIGN: Mailed survey. PARTICIPANTS: 208 patients seen in the University of Utah Asthma Clinic for at least 3 months, age 18 years or older, and on at least 1 asthma medication. MAIN OUTCOME MEASURES: Frequency of pharmacist-provided asthma medication counseling, patients' perception of the importance of counseling, and their satisfaction with counseling. RESULTS: Response rate was 55% (106 of 194 surveys returned). The majority of patients (> 90%) indicated that their community pharmacist "never" or "sometimes" discussed the management of their asthma with them. Frequency of counseling in three predefined areas of asthma education (role of medications, inhaler technique, and prevention of asthma attacks) was 1.91, 1.72, and 1.31, respectively (1 = never, 4 = always). Most patients (76%) considered these areas of counseling to be important for the management of their asthma. Most patients (62%) were "somewhat" to "pretty" satisfied with the type and amount of asthma counseling provided by their pharmacist. Counseling sessions averaged less than 3 minutes. Only 25% of patients would be willing to pay an additional amount for pharmaceutical care. CONCLUSION: Self-reported rates of patient counseling concerning asthma medications in the community pharmacy setting are low and adequately address neither the educational needs of patients nor the recommendations of the National Asthma Education and Prevention Program. PMID- 10467813 TI - Assessing the needs of pharmacists and physicians in caring for patients with epilepsy. AB - OBJECTIVE: To obtain primary care physicians' and community pharmacists' opinions of the Pharmacist Note, a model epilepsy patient profile maintained by the pharmacist and transmitted to the physician as needed, and the information it contains. DESIGN: A cross-sectional descriptive study design was used for collecting and analyzing data. Separate surveys were developed and mailed to physicians and pharmacists. PARTICIPANTS: 554 primary care physicians and 114 community pharmacists in Ohio who interact regularly with epilepsy patients. MAIN OUTCOME MEASURES: Pharmacist and physician opinions on the Pharmacist Note program. RESULTS: Physicians ranked seizure frequency as their most useful piece of information, followed by medication compliance and drug interaction screening. For medication profile and drug interaction screening, most physicians currently use themselves as their primary source of information, although a significant number would prefer to use pharmacists as information sources in these areas (p < .05). A majority (62%) would like to have pharmacists more involved in the care of their patients. Pharmacists identified lack of time and lack of appointments with patients, inadequate pharmacy staff, and insufficient reimbursement as barriers to implementing the Pharmacist Note program. CONCLUSION: Physicians desire pharmacist involvement in specific areas of care for patients with epilepsy, and the feasibility of implementing the Pharmacist Note and similar programs appears promising. However, pharmacists identified barriers to implementation, and these barriers need to be addressed if this type of program is to be successful. PMID- 10467814 TI - Stability and compatibility of tacrolimus and fluconazole in 0.9% sodium chloride. AB - OBJECTIVE: To determine the physical compatibility and chemical stability of tacrolimus injection and fluconazole injection combined in a range of concentrations used clinically for intravenous Y-site co-administration over 3 hours. DESIGN: Tacrolimus injection (Prograf) was combined with fluconazole injection (Diflucan) in volumetric glass flasks at room temperature (23 degrees C to 25 degrees C) to form final concentrations of 5 or 20 micrograms/mL and 500 or 1,500 micrograms/mL, respectively. Immediately after preparation and at 1.5 and 3 hours, triplicate samples and controls were visually inspected and assayed in duplicate by high-performance liquid chromatography; pH of the samples was also determined. SETTING: University of Michigan College of Pharmacy. PATIENTS: Not applicable. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Initial and subsequent concentrations of tacrolimus and fluconazole at 1.5 and 3 hours. RESULTS: More than 99% of the initial concentration of each drug remained in all samples throughout the study period. There was no color change, and the pH of the samples did not change appreciably. CONCLUSION: Tacrolimus and fluconazole are stable and compatible in a range of clinically used concentrations for 3 hours and can be intravenously co-administered through a Y-site. PMID- 10467815 TI - Physical and chemical stability of gemcitabine hydrochloride solutions. AB - OBJECTIVE: To evaluate the physical and chemical stability of gemcitabine hydrochloride (Gemzar-Eli Lilly and Company) solutions in a variety of solution concentrations, packaging, and storage conditions. DESIGN: Controlled experimental trial. SETTING: Laboratory. INTERVENTIONS: Test conditions included (1) reconstituted gemcitabine at a concentration of 38 mg/mL as the hydrochloride salt in 0.9% sodium chloride or sterile water for injection in the original 200 mg and 1 gram vials; (2) reconstituted gemcitabine 38 mg/mL as the hydrochloride salt in 0.9% sodium chloride injection packaged in plastic syringes; (3) diluted gemcitabine at concentrations of 0.1 and 10 mg/mL as the hydrochloride salt in polyvinyl chloride (PVC) minibags of 0.9% sodium chloride injection and 5% dextrose injection; and (4) gemcitabine 0.1, 10, and 38 mg/mL as the hydrochloride salt in 5% dextrose in water and 0.9% sodium chloride injection as simulated ambulatory infusions at 32 degrees C. Test samples of gemcitabine hydrochloride were prepared in the concentrations, solutions, and packaging required. MAIN OUTCOME MEASURES: Physical and chemical stability based on drug concentrations initially and after 1, 3, and 7 days of storage at 32 degrees C and after 1, 7, 14, 21, and 35 days of storage at 4 degrees C and 23 degrees C. RESULTS: The reconstituted solutions at a gemcitabine concentration of 38 mg/mL as the hydrochloride salt in the original vials occasionally exhibited large crystal formation when stored at 4 degrees C for 14 days or more. These crystals did not redissolve upon warming to room temperature. All other samples were physically stable throughout the study. Little or no change in particulate burden or the presence of haze were found. Gemcitabine as the hydrochloride salt in the solutions tested was found to be chemically stable at all concentrations and temperatures tested that did not exhibit crystallization. Little or no loss of gemcitabine occurred in any of the samples throughout the entire study period. However, refrigerated vials that developed crystals also exhibited losses of 20% to 35% in gemcitabine content. Exposure to or protection from light did not alter the stability of gemcitabine as the hydrochloride salt in the solutions tested. CONCLUSION: Reconstituted gemcitabine as the hydrochloride salt in the original vials is chemically stable at room temperature for 35 days but may develop crystals when stored at 4 degrees C. The crystals do not redissolve upon warming. Gemcitabine prepared as intravenous admixtures of 0.1 and 10 mg/mL as the hydrochloride salt in 5% dextrose injection and 0.9% sodium chloride injection in PVC bags and as a solution of 38 mg/mL in 0.9% sodium chloride injection packaged in plastic syringes is physically and chemically stable for at least 35 days at 4 degrees C and 23 degrees C. Gemcitabine as the hydrochloride salt is stable for at least 7 days at concentrations of 0.1, 10, and 38 mg/mL in 5% dextrose injection and 0.9% sodium chloride injection stored at 32 degrees C during simulated ambulatory infusion. PMID- 10467816 TI - Compatibility of gemcitabine hydrochloride with 107 selected drugs during simulated Y-site injection. AB - OBJECTIVE: To evaluate the physical compatibility of gemcitabine hydrochloride (Gemzar-Eli Lilly and Company) with 107 selected drugs. DESIGN: Controlled experimental trial. SETTING: Laboratory. INTERVENTIONS: Samples of 5 mL gemcitabine (as the hydrochloride salt) 10 mg/mL in 0.9% sodium chloride injection were mixed with 5 mL samples of the selected drugs diluted in 0.9% sodium chloride injection or, if necessary to avoid incompatibilities with the diluent, 5% dextrose injection. MAIN OUTCOME MEASURES: Visual examinations of the samples were performed in normal fluorescent light with the unaided eye and using a Tyndall beam (high-intensity monodirectional light) to enhance visualization of small particles and low-level haze. The turbidity of each sample was measured as well. In selected samples, electronic particle content assessment was performed. All of the samples were assessed initially and at 1 and 4 hours. RESULTS: Most of the drugs were physically compatible with gemcitabine hydrochloride during the 4 hour observation period. However, 15 drug combinations had incompatibilities that included color change, increase in haze or turbidity, particulate formation, and gross precipitation: acyclovir sodium, amphotericin B, cefoperazone sodium, cefotaxime sodium, furosemide, ganciclovir sodium, imipenem-cilastatin sodium, irinotecan, methotrexate sodium, methylprednisolone sodium succinate, mezlocillin disodium, mitomycin, piperacillin sodium, piperacillin sodium/tazobactam sodium, and prochlorperazine edisylate. CONCLUSION: Gemcitabine hydrochloride 10 mg/mL admixed in a compatible infusion solution is physically compatible for 4 hours at room temperature with 92 of 107 tested drugs. Simultaneous Y-site administration of gemcitabine hydrochloride with the 15 drugs resulting in incompatibilities should be avoided. PMID- 10467817 TI - Drugs used in acquaintance rape. AB - OBJECTIVE: To describe gamma-hydroxybutyrate (GHB), flunitrazepam, and ketamine and their purported uses to facilitate acquaintance rape. Patient presentation characteristics, treatment regimens, processes to detect the presence of the medications by toxicology screening, and methods to avoid exposure are discussed. DATA SOURCES: MEDLINE search from 1985 to 1998; additional references found within the articles; information obtained from the Internet. STUDY SELECTION: Clinical trials, reviews, and press releases concerning the use of GHB, flunitrazepam, and ketamine to facilitate acquaintance rape. Trials and reviews describing clinical effects, adverse effects, pharmacokinetics/pharmacodynamics, and usage trends were evaluated. Literature judged to be pertinent by the author was included in the discussion. DATA EXTRACTION/DATA SYNTHESIS: Reports of the use of GHB, flunitrazepam, and ketamine in acquaintance rape appear in the medical literature and lay press. Many health care professionals may not be familiar with these medications, and information about caring for patients under their influence is limited. Victims lose their ability to ward off attackers, develop amnesia, and are unreliable witnesses. Because symptoms caused by these agents mimic those of alcohol, not all victims are screened for their presence. Legislative efforts to further limit the use of or access to GHB, flunitrazepam, and ketamine have been initiated at the state and federal levels. Pharmacists should know the symptoms of exposure to the three agents; they should understand treatment regimens, methods to detect the presence of these and other drugs that may have been used in a sexual assault, and techniques individuals can use to avoid becoming victims of drug-assisted acquaintance rape. CONCLUSION: Because of their extensive drug knowledge and frequent access to patients, pharmacists are uniquely positioned to educate patients and other health care professionals about the dangers of acquaintance rape drugs and methods to reduce their risk of becoming victims. PMID- 10467818 TI - Methylphenidate: increased abuse or appropriate use? AB - OBJECTIVE: To address the question of the significant increase in methylphenidate (MPD) prescriptions being written and to make recommendations for health care providers involved in providing care for patients with attention deficit hyperactivity disorder (ADHD) and their families. DATA SOURCES: Medline search 1966-1998 for professional articles using the following search terms- methylphenidate, children, adolescents, abuse; Internet search using MPD, Ritalin, and ADHD; and Paper Chase search using methylphenidate. DATA EXTRACTION: The available literature regarding potential abuse or diversion of MPD consists of case reports, review articles, newspaper articles, and a Drug Enforcement Administration (DEA) publication. All available literature sources were used. DATA SYNTHESIS: Although the media and DEA report significant abuse and diversion of prescribed MPD, a review of the available literature did not reveal data to substantiate these claims. Nonetheless, there are reasons to suspect that abuse and diversion occur. A potential contributing factor to abuse is the reported similarities in pharmacodynamics and pharmacokinetics between MPD and cocaine. Recommendations are made to decrease the possibility of abuse and diversion of prescribed MPD. CONCLUSION: A balanced middle ground must be found regarding the benefits of MPD and its abuse potential. Education of clinicians, patients, and family members is key in ensuring that MPD is used appropriately. PMID- 10467819 TI - Charting the demand for pharmacists in the managed care era. AB - OBJECTIVE: To discuss the future demand for pharmacists and pharmaceutical services in the managed care area. DATA SOURCES: Published reports about the future demand for health professionals and data relating to managed care and the institutional pharmacy work force. DATA SYNTHESIS: A panel from the Pharmacy Manpower Project's Subcommittee to Study Demand Issues examined a broad range of pharmacy-related work-force projections and found widely differing predictions. The panel reviewed recent health care trends related to managed care and data about staff size and changing job numbers in institutional pharmacies to determine which of the predictions were most likely. Medication management problems in the context of increasing prescription numbers and the emergence of data-driven health care support a scenario of a steadily increasing demand for pharmacists and pharmaceutical services. Higher penetration by managed care was not associated with job loss or reduced pharmacy staff size in institutions. CONCLUSION: There is little reason to expect the dramatic downsizing of the pharmacy work force predicted by the third report of the Pew Commission. However, retaining pharmacy roles that are useful to the system and satisfying to pharmacists will require a continuation of current proactive measures by the profession. PMID- 10467820 TI - Compounding botanicals: a legal perspective. AB - OBJECTIVE: To describe the effects of the new law, section 503A, "Pharmacy Compounding," of the Federal Food, Drug, and Cosmetic Act on the compounding of drugs, dietary supplements, and cosmetics. DATA SOURCES: The Food and Drug Administration (FDA) Modernization Act of 1997; Federal, Food, Drug, and Cosmetic Act; Code of Federal Regulations; Federal Register; Food and Drug Administration Guidances. DATA SYNTHESIS: Pharmacy compounding, which traditionally is considered part of pharmacy practice and is regulated by the states, has been confused with drug manufacturing, an activity regulated by FDA. Section 503A, "Pharmacy Compounding," was enacted as part of the Food and Drug Administration Modernization Act of 1997 to distinguish compounding from manufacturing. This law addresses the compounding of drug products, but not necessarily the compounding of botanicals, which may be considered dietary supplements, cosmetics, or drugs. CONCLUSION: Section 503A applies to the compounding of products intended for a drug use, but does not apply to compounded products that are intended for dietary supplement or cosmetic use. Compounded dietary supplements and cosmetics must instead adhere to relevant federal requirements, including those for good manufacturing practices and labeling. PMID- 10467821 TI - The service blueprint as a tool for designing innovative pharmaceutical services. AB - OBJECTIVE: To describe service blueprints, discuss their need and design, and provide examples of their use in advancing pharmaceutical care. BACKGROUND: Service blueprints are pictures or maps of service processes that permit the people involved in designing, providing, managing, and using the service to better understand them and deal with them objectively. A service blueprint simultaneously depicts the service process and the roles of consumers, service providers, and supporting services. Service blueprints can be useful in pharmacy because many of the obstacles to pharmaceutical care are a result of insufficient planning by service designers and/or poor communication between those designing services and those implementing them. One consequence of this poor design and communication is that many consumers and third party payers are uninformed about pharmacist roles. Service blueprints can be used by pharmacists to promote the value of pharmaceutical care to consumers and other decision makers. They can also assist in designing better pharmaceutical services. METHODS: Blueprints are designed by identifying and mapping a process from the consumer's point of view, mapping employee actions and support activities, and adding visible evidence of service at each consumer action step. Key components of service blueprints are consumer actions, "onstage" and "backstage" employee actions, and support processes. DISCUSSION: Blueprints can help pharmacy managers identify and correct problems with the service process, provide pharmacy employees an opportunity to offer feedback in the planning stages of services, and demonstrate the value of pharmaceutical services to consumers. CONCLUSION: Service blueprints can be a valuable tool for designing, implementing, and evaluating pharmacy services. PMID- 10467822 TI - Pharmaceutical care in an integrated health system. AB - OBJECTIVES: An integrated health system in a large metropolitan area, to maximize its manpower and resources, developed a pharmacist-operated Health Management Center (HMC). The primary objectives of the HMC are to provide a continuum of patient care, decrease emergency department visits, decrease episodes of hospitalization, and increase patient satisfaction and quality of life. SETTING: The HMC based at a 300-bed community hospital of an integrated health system in a large metropolitan area. PRACTICE DESCRIPTION/INNOVATION: Chronic diseases, including coagulation disorders, asthma, diabetes, hypertension, congestive heart failure, and dyslipidemia, will be managed by the primary care pharmacist at the HMC. The HMC pharmacist uses a team approach to promote good health by cooperating with patients, the physician clinic director, and other professionals in designing, implementing, and monitoring therapeutic plans that produce specific therapeutic outcomes. The pharmacist evaluates patients using physical assessment skills; performs point-of-care laboratory tests; obtains medication history, including information on compliance, response to drug therapy, and adverse reactions; adjusts and orders medications; and schedules follow-up appointments. INTERVENTIONS: The anticoagulation service was the first program to be established. The pharmacist is authorized to perform point-of-care testing for prothrombin times, adjust doses of anticoagulants, order vitamin K, and schedule return visits per established guidelines. MAIN OUTCOME MEASURES: Emergency department visits, episodes of hospitalization, patient satisfaction, and quality of life. RESULTS: The results for 39 patients 6 months before their enrollment in the HMC's anticoagulation service and for the first 6 months following their enrollment, after adjusting for a 1-month-washout period, showed a decrease in hospitalization rate by 57.9% (p = .078) and total hospital days by 71.1% (p = .108). No change was observed in use of emergency department services. CONCLUSION: The role of the pharmacist at the HMC is a reflection of changes in the health care system that are leading to greater pharmacist involvement in direct patient care. The pharmacist-operated HMC can serve as a model for other hospitals in this and other integrated health systems. PMID- 10467823 TI - Cancer pain management: barriers, trends, and the role of pharmacists. AB - OBJECTIVES: To describe barriers to optimal cancer pain management, activities that have been implemented in recent years to partially overcome these barriers, and the importance of the pharmacist's role in the cancer pain management process. SUMMARY: Cancer pain management remains a formidable challenge. Barriers in the United States include inadequate provider training; patients' reluctance to report pain; and deficiencies within the health care infrastructure, such as restrictive regulation of controlled substances and inadequate insurance coverage. Despite these barriers, recent efforts have raised awareness of the gap between suboptimal and optimal cancer pain management. These include: the development of guidelines (and the conduct of scientific studies) to educate providers about pain management or gaps in the cancer pain management process; regulation that has lessened provider restrictions in prescribing analgesics; the emergence of patient and provider advocacy groups; and the development of pain assessment instruments. Because of their clinical and pharmacologic expertise, pharmacists are crucial to the shaping and success of cancer pain management strategies in the United States today. CONCLUSION: Despite its high prevalence and the increasing awareness of the effects of pain on quality of life, cancer pain management remains secondary to treatment of the disease itself. Refining the cancer pain management process in the United States will depend on continued education and emphasis on pain assessment and a proactive approach to pain management, provider groups' effective collaboration with each other and with patients, and overcoming legal, regulatory, and cost barriers. PMID- 10467824 TI - Finding drug therapy problems. PMID- 10467825 TI - FDA approves fusion protein for treatment of lymphoma. PMID- 10467826 TI - Influenza vaccination increases productivity and reduces misery. PMID- 10467827 TI - Incorporating the Internet into everyday practice. PMID- 10467828 TI - Kangaroo Mother Care. PMID- 10467829 TI - Predictors of breastfeeding in very low birthweight infants at the time of discharge from hospital. AB - In a case-control study carried out in the Kuala Lumpur Maternity Hospital between 1st July 1995 and 31st January 1996 the objectives were (1) to determine the rate of breastfeeding in surviving very low birthweight (VLBW, < or = 1500 g) Malaysian infants following the introduction of the Baby Friendly Hospital Concept, and (2) to identify significant predictors associated with successful breastfeeding in these infants. During the study period, 201 (1.24 per cent) of live-born infants were VLBW infants, 192 (95.5 per cent) were Malaysians, and 141 (73.4 per cent) of them survived to go home. The breastfeeding rate among all surviving VLBW Malaysian infants at the time of discharge was 40.2 per cent (57/141). The mothers of 126 (89.4 per cent) VLBW Malaysian infants were interviewed before discharge. Logistic regression analysis showed that, after controlling for various confounders, the significant predictors associated with successful breastfeeding were: (a) Malay mothers (odds ratio: 6.0; 95 per cent CI: 1.9, 19.4), (b) mothers with educational levels of between 7 and 9 years (odds ratio: 3.6; 95 per cent CI: 1.0, 12.2), and (c) earlier age of commencement of enteral feeds in the VLBW infants (for each additional day delay in commencement of feeding, odds ratio of breastfeeding was 0.5; 95 per cent CI: 0.4, 0.8). PMID- 10467830 TI - Rickets in Nigerian children: response to calcium supplementation. AB - In a previous study of rachitic children in Jos, Nigeria we concluded that inadequate dietary intake of calcium was the primary contributing factor to the development of their rickets. The objective of the present study was to determine the effect of calcium supplementation in 10 children with radiographically and biochemically proven rickets from the same geographical area. Rachitic children were provided with calcium supplements of 1000 mg/day for a period of 3 months. Serum and urine samples were obtained at baseline and at 24 hours, 1 week, 4 weeks, and 12 weeks after initiation of supplementation. Serum calcium, phosphorus, alkaline phosphatase, intact parathyroid hormone, 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D were measured at each time point. Dietary recalls obtained at two separate times were used to estimate usual daily intakes of calcium and phosphorus. Ten non-rachitic age-matched controls from the same geographical area were recruited for comparison. Nine of 10 rachitic subjects had radiographic evidence of healing after 3 months of calcium therapy. Although serum calcium concentrations returned to control levels, other biochemical data indicated that the rickets of these subjects may have been multifactorial in aetiology, pointing to a possible defect in the synthesis of 25-hydroxyvitamin D. PMID- 10467831 TI - Effective health education in rural Gambia. AB - We aimed to measure the impact of a structured health education programme on the incidence of infectious disease in young children of a rural Gambian village. In a prospective intervention study, mothers of children under 3 years living in a rural West African village without running water (population 1600) were divided into two groups by site of residence. Group 1 (n = 70) received a fortnightly education talk on hand-washing and skin hygiene, and group 2 (n = 84) on family planning. The incidence of infectious skin and diarrhoeal, eye, and respiratory diseases were recorded at the clinics attended by all children. The mean incidence of skin infections in the children of mothers of group 1 (1.07) was significantly lower (p < 0.05) than that of group 2 (1.59) during the rainy season. Between 1981 and 1994 the incidence of skin, diarrhoeal, and eye disease showed a statistically significant (p < 0.0001) decline following the implementation of the health education programme. There was no statistical significance in the trend for respiratory disease, the incidence of which would not be expected to be affected by hygiene education. Focused health education had a sustained beneficial effect on the health of young children living in conditions where infectious diseases and poor hygiene are common. PMID- 10467832 TI - Helminthic infection and anthropometric indicators in children from a tropical slum: Ascaris reinfection after anthelmintic treatment. AB - We evaluated helminthic infection and anthropometric indicators of nutritional status in a group of school-age children from a slum area of Caracas, Venezuela. The proportions of children at or below the 10th percentiles for height/age and weight/age were significantly higher in those presenting helminthiasis than in those uninfected. Although this could partially reflect a codependence of both helminthic infection and undernutrition or poverty, when the children were administered regular anthelmintic treatment for a year their anthropometric values improved significantly. When they were re-evaluated 8 months after the end of anthelmintic administration, the degree of reinfection by the most common helminth, Ascaris lumbricoides, was significantly higher in the growth-retarded children. These results confirm the relationship between helminthic infection and decreased growth rates in underprivileged populations, and indicate that children at nutritional risk are more susceptible to such infections, even after a prolonged parasite-free period. PMID- 10467833 TI - The pattern for common anaemia among Saudi children. AB - Anaemia is of frequent occurrence in children in different parts of the world and poses a significant health problem. A few isolated reports indicate that anaemia occurs at a high prevalence rate in Saudi Arabia though the actual prevalence in several regions is not known. The aim of the present study was to determine the prevalence of different types of anaemias in Saudi children in different areas of the country. Blood samples were collected from 5381 children less than 14 years of age, and haematological analysis and red cell indices were determined. The results of haematological parameters were used to group the children as anaemic (Hb < 11.2 g/dl) and non-anaemic (Hb > 11.2 g/dl) and the red cell indices were used to classify the anaemia as hypochromic-microcytic, normochromic-normocytic, and normochromic-macrocytic. The overall prevalence of anaemia in Saudi children was 24.8 per cent. The prevalence was highest in the children from the Eastern province (41.3 per cent) and lowest in the central province (16.5 per cent). Within each province differences were obvious in the prevalence of anaemias in the different areas. The majority of the anaemia in the eastern and south-western provinces was hypochromic-microcytic, while in the north-western and central provinces normochromic-normocytic anaemia occurred most frequently. Macrocytic anaemia was not encountered in any of the screened areas of the central province and many areas of the eastern province. However, in north-western and south western provinces it occurred at a frequency of 0.15-3.4 per cent. The data show that anaemia is a frequent problem in Saudi children living in different parts of Saudi Arabia and emphasizes the need for nutritional and genetic assessment to determine the nutritional contributions to anaemias and hence the correction of nutritional anaemias by proper dietary intervention. PMID- 10467834 TI - Prognostic value of serum C-reactive protein in kala-azar. AB - The currently recommended protocol for treatment of kala-azar (KA) necessitates repeated bone marrow/splenic aspiration to monitor the response and duration of therapy as well as to detect resistance and change to alternative drugs. These procedures being invasive, there is a pressing need for less invasive diagnostic tools for this purpose. We have evaluated the role of C-reactive protein (CRP) in 201 children with visceral leishmaniasis at different stages of the disease to work out the relationship, if any, between CRP levels and disease activity, including response to therapy. The subjects belonged to the 2-12 year age group in whom CRP estimation was done on admission, every 5th day during treatment, and repeated on follow-up at 2 and 6 months. The levels were compared with those of 50 randomly chosen age-matched healthy children who served as controls. The mean serum CRP value in the study group before the commencement of treatment was 62.96 +/- 1.03 mg/l, which was significantly higher (p < 0.001) in comparison to the control group. Commencement of treatment resulted in a simultaneous decline in serum CRP. Parasitic clearance from the spleen was faster in patients with an initial low serum CRP level (< 60 mg/l) in comparison to patients with high levels (> 60 mg/l). During treatment, mean serum CRP levels were significantly higher in late responders than in early responders (p < 0.001). Correlation of CRP levels to indicate the presence or absence of parasites suggested a cut-off level of 12 mg/l by day 10, with a sensitivity of 82.5 per cent, specificity of 78.5 per cent, positive predictive value of 92 per cent, and negative predictive value of 60.2 per cent. Our observations suggest a promising role for CRP estimation every 5-10 days during therapy in visceral leishmaniasis for monitoring the response to therapy and to detect possible resistance. PMID- 10467835 TI - Term infants with hypoxic ischaemic encephalopathy: poor neurodevelopmental outcome despite standard neonatal intensive care. AB - Perinatal asphyxia may be severe and affect the neurological system, causing hypoxic ischaemic encephalopathy (HIE). Twenty-three term infants with HIE were studied over 3.5 years. Nine died and of the 14 survivors, eight were normal and the rest had abnormal neurological sequelae. Favourable prognostic markers were a 5 minute Apgar score > 5, first pH > 7.1, and stage I HIE. Despite standard neonatal intensive care management, the mortality and morbidity rates for these infants remained high. PMID- 10467836 TI - Primary staphylococcal pneumonia in young children: a review of 100 cases. AB - Staphylococcus aureus (S. aureus) is responsible for a small proportion of acute respiratory infections in children. Nevertheless a high index of suspicion is required because of the potential for rapid progression, the need for antibiotics different to those routinely administered in the treatment of pneumonia, and the high incidence of complications. There are few data from developing countries. The objective of this retrospective review was to document the natural history of primary staphylococcal pneumonia at Red Cross Childrens' Hospital in Cape Town over a 7-year period (1989-1995). Staphylococcal pneumonia was defined as acute pneumonia with microbiological evidence of S. aureus or with characteristic radiological features. One hundred patients were identified. The median age was 5 months, 78 patients being below one year of age. Cough and fever were present in almost all patients at the time of presentation. Tachypnoea, recession, dullness, and crepitations were commonly elicited signs. Initial chest radiographs revealed empyema, pleural effusion, or pyopneumothorax in 67 patients. A further 26 patients developed such changes on subsequent chest radiographs. Pneumatocoeles were identified in 37 patients--most of these were only noted on radiographs taken some days after admission. Microbiological confirmation was obtained in 92 cases. S. aureus was isolated in 23/98 blood cultures, 62/67 pleural aspirates, and from tracheal aspirates in 16 cases. Intercostal drains were inserted in 67 cases and 20 children underwent thoracotomy. The case fatality rate was 7 per cent. This study shows that primary staphylococcal pneumonia is chiefly a disease of infants. Symptoms and signs were similar as for other forms of acute pneumonia, although in the majority of cases chest radiographs taken at the time of admission suggested the diagnosis. Treatment with antibiotics and drainage of empyema resulted in a good outcome in the majority of cases. PMID- 10467837 TI - Intraventricular haemorrhage and its prognosis, prevention and treatment in term infants. AB - The purpose of this study was to investigate the prognosis of intraventricular haemorrhage (IVH) in term infants and its prevention and treatment. The authors diagnosed IVH in full-term newborns by using computerized tomography (CT) or cranial ultrasonography (US). The results of CT or US were reviewed and the grade of haemorrhage was determined by an independent radiologist using Papile's criteria. All the infants were examined on the first day of their life by a paediatrician and judged to be full term. Survivors were examined between age 2 and 10 years at the Child Development Clinic by a Developmental Paediatrician using the Gesell scales and a standard neurologic examination. The results showed that three of 36 infants (8 per cent) died; complications of pregnancy were present in 17 mothers (47 per cent); nine women tested negative for platelet antigen 1 and their infants exhibited alloimmune thrombocytopenia. Age at diagnosis ranged from in utero to 28 days. Clinical presentation included feeding intolerance, irritability, jaundice, fever, and restlessness. Of the nine children with grade IV IVH, three died and six survivors were severely handicapped. Overall, 22 (67 per cent) of 33 survivors had no or mild handicap. The results of this study suggest that severity of haemorrhage was of prognostic value. Perinatal alloimmune thrombocytopenia turned out to be the single most important cause of severe haemorrhage and poor outcome. Identification and treatment of these infants must begin in utero if we are to prevent IVH and its complications in this group of patients. PMID- 10467838 TI - Intestinal parasites in Malaysian children with cancer. AB - In this prospective study, we examined stool specimens from children with cancer receiving chemotherapy who were admitted for fever to the Universiti Sains Malaysia Hospital in Kota Baru, Kelantan. Stool specimens were examined for ova and cysts of parasites. Over a period of 15 months, there were 129 febrile episodes in 50 children with cancer and, in all, 237 stool specimens were examined. Sixty-six per cent of febrile episodes were associated with neutropenia and 9 per cent were associated with diarrhoea. Stool parasites were found in 42 per cent of children. The most common were helminths, followed by protozoa. Trichuris trichiura was the most common parasite (24 per cent), followed by Ascaris lumbricoides (22 per cent). Hookworm was found in 2 per cent. Giardia lamblia was found in 6 per cent of children, Blastocystis hominis in 4 per cent, and Cryptosporidium parvum in 2 per cent. PMID- 10467839 TI - Cure of beta-thalassaemia major by umbilical cord blood transplantation--a case report of Malaysia's first cord blood transplantation. AB - A 25-month-old boy with beta-thalassaemia major was presented with an opportunity for umbilical cord blood transplantation when his unborn sibling was diagnosed in utero to be a beta-thalassaemia carrier and also human leucocyte antigen compatible. A barely adequate amount of cord blood was collected at the birth of his sibling and infused into the patient after appropriate chemo-conditioning. Engraftment occurred without major complications. The subject is now alive and well 9 months post-transplant, thus marking our first success in umbilical cord blood transplantation. PMID- 10467840 TI - Adenosine deaminase in childhood pulmonary tuberculosis: diagnostic value in serum. AB - The diagnostic value of serum adenosine deaminase (ADA) activity was evaluated in childhood pulmonary tuberculosis. Serum ADA levels were measured in 20 children diagnosed with pulmonary tuberculosis (group 1) and 150 children (group 2) including 128 with tuberculosis infection (Mantoux test positive) and 22 healthy children. In group 1, the mean serum ADA activity was 74.06 +/- 18.5 U/l, which was significantly (p < 0.001) higher than that of group 2 (40.36 +/- 12.0 U/l). A serum ADA level of > or = 53.76 U/l had a sensitivity of 100 per cent, specificity of 90.7 per cent, positive predictive value of 58.8 per cent, and a negative predictive value of 100 per cent in children with tuberculosis disease. To conclude, measurement of serum ADA activity was a useful diagnostic tool in childhood pulmonary tuberculosis. PMID- 10467841 TI - Classification trees and logistic regression applied to prognostic studies: a comparison using meningococcal disease as an example. AB - The authors used logistic regression and classification trees to develop prediction models for fatal outcomes in meningococcal disease in a cohort of 829 children hospitalized for meningococcal disease during 1989-1990 in Rio de Janeiro. The area under the receiver operator characteristic (ROC) curve was 92 per cent for logistic regression and 88 per cent for classification trees. Logistic regression may be preferred when the main objective is to obtain explicit measures for statistical inference and measures of the force of the association between each variable and the outcome. However, estimation of the probability of dying for each patient involves manipulation of the logistic regression formula, which would not easily be done in an emergency room. Classification trees provided comparable discrimination between fatal and non fatal outcomes, and yielded a graphical display of the results that is easier to understand and is straightforward to apply in clinical settings. PMID- 10467842 TI - Relationship between CRIB score and intraventricular haemorrhage in very low birthweight infants. PMID- 10467843 TI - Acute gastroenteritis caused by E. coli O.157:H7 in Pakistani children. PMID- 10467844 TI - [Occupational medicine practice in Switzerland in the dawn of the year 2000]. PMID- 10467845 TI - [Alcohol, drugs and the workplace]. PMID- 10467846 TI - [When work organization becomes the source of pathologies]. PMID- 10467847 TI - [Can practising physicians be industrial physicians?]. PMID- 10467848 TI - [Contribution of the work inspector physician to the protection of workers' health in the Neuchatel and Geneva cantons]. PMID- 10467849 TI - [Analysis of the health risks in the workplace. Contribution of the work hygienist]. PMID- 10467850 TI - [Industrial health education in the Neuchatel canton]. PMID- 10467851 TI - [Biological surveillance: its utility for the evaluation of chemical risks]. PMID- 10467852 TI - [Tuberculosis screening: computer program development for the interpretation of the Mantoux reaction in personnel at the Geneva University Hospitals]. PMID- 10467854 TI - [Skin diseases related to work: how to approach them?]. AB - Work-related skin problems are frequent and mainly affect the hands. They can be approached by looking for indications of the 3 principal underlying types of dermatitis: irritant, allergic, or chronic. Irritation and contact allergy are closely associated: the irritation facilitates the flowering of the allergy. Skin irritation due to working conditions is often multi-causal and repetitive. Cutaneous atopy--but not that which is only respiratory or mucosal--is the main genetic risk factor for the development of a work-related skin disease. The characteristics of irritation and allergy tend to merge when the lesions become chronic. An 8-step process is suggested to establish the causal relationship between potentially damaging substances in the workplace and the resulting skin problems. In Switzerland, professional dermatitis falls under the purview of the accident insurance laws. These laws recognize causal responsibility whenever the work substances or activities are preponderant over non-professional causes. PMID- 10467853 TI - [Management of professional exposures to blood and other biological liquids contaminated with hepatitis B and C and HIV]. PMID- 10467855 TI - [The prison physician: simple institutional medicine?]. PMID- 10467856 TI - [The practice of psychiatry in the prison milieu in the Vaud canton]. PMID- 10467857 TI - [The physician's management of a patient who is also a prisoner]. PMID- 10467858 TI - [The status of the forensic biological service in the Russian Federation in 1997]. AB - The main parameters of the activities of forensic biological departments are analyzed for the year 1997. The staff, number of experts, number of expert evaluations and objects of examination, and work load per staff expert are presented. Shortcomings in the activities of departments are enumerated and recommendations for improving the quality of expert evaluations are offered. PMID- 10467859 TI - [The determination of haptoglobin in specimens of biological origin]. AB - Presents the genetic aspects of haptoglobin in health, the rare types, and shifts of Hp fractions resultant from changes in the redox function (particularly in children) caused by high temperature, carbon monoxide, oxygen insufficiency, etc. PMID- 10467860 TI - [The tissue survival problem in using current methods of morphological study]. AB - Time course of ultrastructural changes and RNA production in human brain and myocardial cells during the period between clinical and biological death is studied. Changes in the cytoplasm of brain neurons develop 4 hours after death, and 8 h post mortem the destructive changes in the cytoplasm and nuclei progress. During the period of 4-13 h post mortem the endotheliocyte nuclei are capable of producing RNA. Active production of RNA is on-going in the myocyte nuclei 4 h after death; the synthetic activity decreases by 8 h and by 13 h the synthetic activity in autopsy material is over. PMID- 10467861 TI - [The contour characteristics of the mucous membrane of the hard palate as dependent on the form of its osseous base]. AB - Analyzes the relief of the hard palate mucosa, depending on the shape of osseous base. Presents data on the prevalence of various morphological shapes of lateral folds in the hard palate mucosa and the levels of their disposition in the ipsiloid, ellipsoid, and parabolic osseous base of the hard palate. Demonstrates the possibility of using the characteristics of the hard palatal mucosa relief as an extra test for personality identification by the oral status. PMID- 10467862 TI - [The role of certain sections of the central nervous system in the genesis of sudden death]. AB - Hypothalamic and hippocampal lipid composition in sudden death from coronary heart disease is studied. Thin-layer chromatography was the main method of biochemical analysis. Disorders in the lipid component of hippocampal cytostructures involved mainly the cholesterol fractions. Metabolic disorders in the hypothalamus involved the phospholipid metabolism. The above-mentioned biochemical changes were paralleled by ultrastructural abnormalities. Analysis of the data demonstrates a probable metabolic basis of a central mechanism of sudden cardiac death. PMID- 10467863 TI - [Blood and liver monoamine oxidases in fatal alcoholic intoxication]. AB - Biochemical studies of monoamine oxygenases (MAO) were studied in albino rats and in subjects dead from ethanol poisoning, alcoholic cardiomyopathy, and coronary heart disease. Experiments demonstrated that chronic alcoholization leads to increase in the levels of both types of MAO (A and B), the longer the alcoholization, the more pronounced the increase. Intake of high ethanol doses in the course of regular alcohol abuse involves a drop in MAO activity (by 1.9-2-2 times). Study of MAO helps detect chronic alcoholization and differentiate it from coronary heart disease and alcoholic cardiomyopathy and thus make an objective conclusion about the cause of death and thanatogenesis. PMID- 10467864 TI - [The methodological aspects and prospects of using dynamically modified sorbents for the analysis of biological specimens by means of high-performance liquid chromatography]. AB - Published reports and authors' experience gained in high-performance liquid chromatographic analysis of dynamically modified adsorbents are reviewed. The advantages of this variant of liquid chromatography are shown: the method is economic and more effective for analysis of mixtures of a complex composition, including biological objects. Examples of mycotoxin, morphine, and other toxicologically significant agents are offered. The method is recommended for wide use in forensic medical expert evaluations. PMID- 10467865 TI - [The determination of clozapine in blood and urine by gas chromatography-mass spectrometry]. AB - A method for measuring closapine in the blood and urine by gas chromatography mass spectrometry as a trifluoroacetic derivative is proposed. The threshold level for closapine detection is 25 ng/ml in the blood and 30 ng/ml in the urine. Calibration curves are linear in the range 0.025-5 mcg/ml for the blood and 0.03 50 mcg/ml for the urine. The method can be used in forensic chemical and clinical toxicological analysis. PMID- 10467866 TI - [The gas chromatographic detection of acetylene in cadaveric material]. AB - Acetylene traces were detected by gas chromatography in the cadaveric right crural muscle of a 30-year-old man dead from an explosion of an acetylene reservoir at a plant. Acetylene was identified using the absolute calibration method on 3 standard gas chromatographic columns, reaction gas chromatography, and acetylene "deduction" by silver sulfate on silicagel. PMID- 10467867 TI - [Synthetic pyrethroids: their nomenclature and properties]. PMID- 10467868 TI - [A case of acute heroin poisoning]. PMID- 10467869 TI - [An unusual case of trauma to the passenger in the interior compartment of a motor vehicle]. PMID- 10467870 TI - [Everything was done--patient still not satisfied: introduction to evidence-based surgery]. AB - Although the best knowledge in academic medicine has been used, some patients are still unsatisfied. By reason of these experiences there is a certain need to reflect our patterns of thinking. The surgical act is based on the laws of causality from Newton and Galilei and it is determined by the principle of cause and effect. Therefore the healing process must be interpreted as a turning back of pathogenesis or the linear chain of causalities. Scientific knowledge of this century demonstrate, that biologic healing processes are connected with the laws of cybernetics and the principles of semiotics. There are functional relations between the level of the organ (impairments) of the individual (disabilities) and of the society (handicaps). This International Classification of Impairments, Disabilities and Handicaps (ICIDH) serves as the key for the management of chronic diseases. An independent and a separate classification of the severity in each level is necessary to identify the consequences of the disease to the patient. Surgical interventions occur on the organ level, benefit and evidence are reflected especially on the individual level (gain of abilities). The assessment and the integration of the so called "sensory impairment", influenced by the biographical events of the patient, an the evaluation of the psychosocial resistance are important factors to recognize a unfavourable conformity between the degree of impairment and the degree of disability. With this classification a more patient-oriented discussion of the indications regarding operative procedures can be realised. The MARA model (mean age related ability) serves as a pragmatic basis for the description of the benefits of carried out and omitted interventions as changes of abilities by using the MARA curve as ethical guideline. This model, which is derived on ICIDH, the hierarchy of needs and the salutogenesis (semiotics, cybernetics), facilitate the introduction of evidence based surgery. It helps to estimate the several predictive values and correlation factors influencing the manifestation of the disease. On this way astonishing results in evidence can be expected. Finally many misunderstandings in health care discussions are explained by the fact that the differences between pathology and illness are not clearly interpreted. PMID- 10467871 TI - [Can psychosomatic concepts make a contribution in surgery?]. AB - All medical reasoning and action is based on a medical theory. Johannes Muller, the renowned 19th century physiologist, stated: "What we experience with our senses is not a mirror of objects characteristics, but rather depends on the living organism's interpretation of sense-receptor stimulation." A mechanistic model of medicine which does not reflect on and include individual reality and the influence of personality and personal history subjects patients to the danger of surgery; psychosomatic concepts reduce this danger. If the surgeon integrates (not adds!) the psychological and social dimensions in his theory of medicine, then the question of what psychosomatic concepts can contribute to surgery is answered: A patient with atypical facial pain due to a conversion reaction will not be operated. Patients will be dismissed and return home earlier after hip joint operations. Persons with a strong body fixation and pain masking their fantasies of invulnerability (narcissistic block) will not be subjected surgery. Women with abdominal pain connected with childhood sexual abuse will not be submitted to operations in the genital area etc. PMID- 10467872 TI - [Psychological aspects in surgery--surgical aspects in psychology: psychosomatic medicine in routine practice?]. AB - Medicine appears to have reached a standstill, in particular surgery: While progress in apparative diagnostics and therapy, e.g. by ultrasound, endoscopy, laparoscopy, and arthroscopy, has reached in some areas "breathtaking speed", the parties concerned-surgeons and patients-lose in reserve air to breathe: no time for any profound consideration, no chance for in-depth talking before surgical procedures are performed, no training for giving real support after surgery. Do we see a brand of surgery which feels responsible only for a human machine without soul? Meanwhile discussions have begun inside medicine in general about a change of paradigms, ways of thinking, the model of the world. Surgery can even make substantial contributions to this change of paradigms. Surgery for the human machine? Psychosomatic ways of thinking are first seen not applicable for surgery. Substantial anxiety exists to take notice of each other. Three concrete examples from everyday practice in surgery demonstrate, how psychosomatic thinking can change and enrich surgical practice: From the area of indications reflections about appendectomy, from the operative-surgical area the phenomenon of self-destructive behaviour, from the restitutive area experiences in treatment of osteomyelitis. Back to the interpersonal area! Psychosomatics must be recognised as a way of thinking and be integrated as such into surgery. A so called "integrated surgery" will arise. The examples demonstrate also, that it is not some anonymous surgical medicine, which must and can change, but that only the individual surgeon as a concrete person can bring back his work into the interpersonal area. PMID- 10467873 TI - [Evidence-based medicine: modern scientific methods for determining usefulness]. AB - For quite some time, clinical epidemiology has introduced the art of critical appraisal of evidence as well as the methods of how to design sound clinical studies and trials. Almost unnoticed by most medical institutions a new hierarchy of evidence has emerged which puts well thought out trials, able to document unbiased treatment benefit in terms of patient suffering, above pathophysiological theory. Many controlled trials have shown, in the meantime, that the control of laboratory or other kind of pathologies and the correction of anatomical abnormalities do not necessarily mean a benefit for the patient. Concepts relating to this dissection of evidence include: Surrogate fallacy ("cosmetics" of laboratory results or ligament or cartilage "cosmetics" in surgery), confounding (spurious causal relationships), selection bias (comparison with selected groups) as well as lead-time bias (mistaking earlier diagnosis as increase of survival), length bias (overlooking differences in the aggressiveness of diseases as determinants of disease stage distributions) and overdiagnosis bias (mistaking the increasing detection of clinically silent pathologies as improvement of prognosis). Moreover, absolute instead of relative risk reduction needs to be used to measure patient benefit. The incorporation of decision analysis and of the concepts or clinical epidemiology will improve the efficiency and quality of medicine much more effectively than the sole focus on technical medical performance. Evidence based medicine is the systematic and critical appraisal of medical interventions, based on the understanding how to avoid the fallacies and biases mentioned. PMID- 10467874 TI - [The path from science to the practicing surgeon. Engagement of documentation of the Swiss Academy of Medical Sciences for providing evidence-based medicine]. AB - The flood of information that comes along with the rise of electronic media has changed the expectations towards the Documentation Service of the Swiss Academy of Medical Sciences (DOKDI): Evidence Based Medicine (EMB) in particular not only demands procurement of information, but also a selection regarding quality and relevance: The question arising out of the clinical situation requires an answer correct in its content and helpful in the specific situation. Getting an idea of what evidence exists about the correctness of a certain procedure through critical lecture is an ideal often obstructed by lack of time and methodical problems in the practice; therefore, one often has to rely on evidence acquired through others and consult e.g. the Cochrane Library. DOKDI commits itself to the development of systematic reviews as well as to the dissemination of evidence found by using its experience in the documentation with electronic media and by providing the corresponding infrastructure. In addition to these activities, the Academy has spoken a grant for the training of EBM-Tutors. During a weekly workshop held in Oxford, clinicians will be trained as EBM-Tutors. This will allow an increasing number of EBM-Workshops held in Switzerland in the future. PMID- 10467875 TI - [Distal biceps tendon ruptures--experiences with soft tissue preserving reinsertion by bone anchors]. AB - The repair of the distal biceps tendon rupture with bone anchors achieves excellent results. The important function of the M. biceps brachii as a powerful supinator and flexor can be reestablished and the soft tissue can be protected. We describe the repair of the rupture of distal biceps tendon by an anatomic refixation of the tendon to the tuberositas radii with bone anchors through a single anterior incision. In a four year period we treated eight patients with this technique. All patients were men with a mean age of 50 years. In each case the mechanism of injury was a single traumatic event with a large load on the flexed elbow. In average surgery was performed within five days and the patients stayed in the hospital for two days. Clinical follow up after almost two years showed no nerve damage or heterotopic bone formation. The range of motion and isometric muscle testing were almost symmetrical. All patients were able to return to preinjury activity level without any limitations. PMID- 10467876 TI - [Drug provision of the Russian Federation's population and optimal measures for its improvement]. PMID- 10467877 TI - [Immunology of tuberculosis: current status]. AB - The paper outlines the high-priority trends in basic and applied investigations of antituberculous immunity. It presents the results obtained from studies of the genetic mechanisms responsible for the control of susceptibility in experimental tuberculosis, from analyses of novel chemical and recombinant tuberculosis vaccines, from examinations of the role of immunological memory in the disease. While describing the results of tuberculosis immunodiagnosis, particular emphasis is laid on its major aspects: on the differential diagnosis of tuberculosis and other lung diseases and on the screening of high-risk group populations for disease progression in order to made further in-depth studies. Current aspects of evaluating the immune status and nonspecific responsiveness in patients with tuberculosis are considered. PMID- 10467878 TI - [Clinical and immunological studies in borderline mental disorders: problems and solutions]. AB - The study demonstrated that patients having an acute response to stress and those with adaptative disorders with prevailing short-term depressive disorders had certain immunological disorders. Therapy with tranquilizers indicated that they produced an inhibitory effect on T- and B-cell immunity, antibody formation and lymphokine production. Among the immunomodulating peptides myelopid, licopid, and Tactivin, the former was found to be the most effective against anxiety and fear while licopid was the most effective agent in the treatment of psychoasthenic disorders. The findings suggest that therapy with psychotropic peptides will open new vistas in treating mental disorders. PMID- 10467879 TI - [Heat shock proteins: new prospects of myocardial cytoprotection]. PMID- 10467880 TI - [Current views on molecular structure of benzodiazepine receptors]. AB - It is generally accepted that gamma-aminobutyric acid (GABA) is the most important inhibitory transmitter. It interacts with specific receptors. In the central nervous system of the vertebrates there are three types of GABA receptors: GABAA, GABAB, and GABAC receptors. Channel gating at GABAA receptors allosterically is modulated by a wide of neuroactive compounds, including benzodiazepine ligands. The structural and functional bases for the heterogeneity of benzodiazepine receptors are discussed. Investigations of the functional properties of GABAA and benzodiazepine receptors may contribute to progress in neurobiology, neuropathology, neurochemistry, toxicology, and pharmacology. PMID- 10467881 TI - [Homeostasis of small molecules originating from microbes and its role in microbial relations with the host]. AB - It is suggested that disorders at the low molecular levels--at the level of small molecules originating from microbes (SMOM) underlie nonspecific pyoinflammatory diseases and decompensation of SMOM homeostatic disorders in the human blood plays a certain role in the complex multifactorial pathogenesis of sepsis. The blood from 16 donors showed fatty acids (hydroxy acids, branched, short-chain, unsaturated, cyclopropanic acids), aldehydes, alcohols, and phenylcarbolic compounds which are not produced by mammalian cells and which are structural components or microbial metabolites. That from 59 patients with various diseases (peritonitis, endocarditis, urinary tract infection, etc.) displayed a reduction or a complete disappearance of only small molecules which are as part of the endogenous microflora and, concomitantly, a 100-fold increase or more in other molecules, and the emergence of new SMOM lacking in the donor blood. The findings yield a concept that there is the homeostasis of small molecules originating from microbes, whose severe disorders (SMOM homeostatic decompensation) are likely to be a first link in the genesis of a systemic inflammatory response, in the development of septic shock and multiorgan failure. PMID- 10467882 TI - [Laws of typological interactions of clino- and antiorthostatic systemic and cerebral circulation in children]. AB - Based on P. K. Anokhin's systems approach, the authors analyze the typological features of the mechanisms of clinostatic and anti-orthostatic systemic and cerebral circulation in children aged 7-9 years. Children with eukinetic systemic circulation have stable total peripheral vascular resistance in antiorthostasis, the mechanisms of cerebral hemodynamic regulation ensure constriction of local arterioles, those with hypokinetic systemic circulation have decreased total peripheral vascular resistance and lower constriction of cerebral arteries and arterioles. In hyperkinetic circulation, total peripheral resistance increases with slight constriction of cerebral arterioles and marked deterioration of venous blood outflow from the cerebral pool. The typological features of systemic and cerebral circulation should be borne in mind in pediatric clinical practice. The findings allow one to mathematically predict the changes occurring in anti orthostatic hemodynamics in the context of its typological features. PMID- 10467883 TI - [Synaptic structural organization as a determinant of selective sensitivity and plasticity of brain neurons in postanoxic period]. AB - The neuro- and synaptoarchitectonics of the sensoty-motor cortex of the brain and cerebellum was comparatively analyzed in an experiment on adult albino rats. There was a relationship between the formation of axospinal synapses to the type of organization of paramembranous cytoskeleton. A role of cytoskeleton's factors of synaptic contacts in the pathogenesis of postanoxic selective neuronal lesion and in the compensatory reorganization of synaptoarchitectonics is hypothesized. PMID- 10467884 TI - [Clinical and immunochemical characteristics of remittent multiple sclerosis]. AB - Immunochemical monitoring was used to examine 37 patients with remittent multiple sclerosis (MS) for 3 years. There were changes in the blood concentrations of the supernatants IL-1 beta, IL-2, IL-6, alpha-TNF, in the serum levels of the adhesion molecule sVCAM, the neurospecific proteins GFAP and alpha 2GP. Only does active MS show a simultaneous increase in the serum concentrations of SVCAM and GFAP and alpha 2GP, which may be used as a test for the objectification of MS aggravation, the efficiency of therapy. The revealed higher secretion of antiinflammatory of cytokines at all disease stages, no recovery of blood-brain barrier responsiveness and at remission provide evidence for the likelihood of a permanent immunopathological process in MS with prevailing inflammatory and destructive changes during a clinical exacerbation and of the induction of active astrogliosis at its remission. PMID- 10467885 TI - [Electron microscopic and mass spectrometric studies of lymphedema pathogenesis aspects]. AB - A comprehensive examination was made, which included histological, electron microscopic and chromatographic mass spectrometric studies of biological specimens obtained from 18 patients with leg lymphedema. The development of lymphedema was found to accompanied by the specific connective tissue structure being formed along with the presence of low-differentiated and necrotic areas associated with the cytopathology of blood and lymph vessels and impaired microcirculation. The connective tissue areas under study were demonstrated to have a pronounced antigenicity and aggressive properties against other tissues and structures. The morphological changes revealed are directly related to the impaired metabolism of protein, fatty acids, and complex polysaccharides. PMID- 10467886 TI - [Cytogenic characteristics and virogenic status of lymphoid cells from malignant baboon lymphomas]. AB - Four lymphoid cell cultures obtained from Papio hamadryads with malignant lymphomas were studied. The cultured cells produced two lymphotropic EBV-like herpes virus of Papio (HVP) and T-lymphotropic monkey retrovirus STLV-1 (HTLV-1 family) or HVP alone. More than 100 subcultured cell lines were shown to remain mixed B- and T-cellular, the levels of T cells of different subpopulations were 15-50%. Cytogenetic investigations showed that one of the 4 cultures was tumorogenic and the others were derived from the normal cells of monkeys with malignant lymphomas. It is suggested that the presence of a cultural virus (viruses) may affect the karyotypes of cells. PMID- 10467887 TI - [Role of socio-hygienic science in reforming national public health]. PMID- 10467888 TI - Response covariation. The relationship between correct academic responding and problem behavior. AB - This article examined the relationship between the accuracy of academic responding and aggression for two boys with mild mental retardation. Their teacher reported low rates of correct responding and high rates of aggressive behavior during spelling instruction. A functional analysis showed that aggression was escape maintained. Following the functional analysis, participants were tested on relations between printed, photographic, and dictated stimuli corresponding to their spelling words. On pretests, they were unable to match printed words to their photographs or to their dictated names; they could neither name the printed words nor spell the photographs or dictated words. High rates of aggression were observed during the pretests. The participants then were taught the letter-by-letter construction of the appropriate words when shown photographs. On posttests, the participants correctly matched printed words to their photographs and dictated names. In addition, they correctly named printed words and spelled dictated words orally. Data showed that rates of problem behavior negatively covaried with improvements in the participants' academic responding. PMID- 10467889 TI - Using rapidly alternating multiple schedules to assess and treat aberrant behavior in natural settings. AB - The validity of using rapidly alternating multiple schedules (RAMS) as a method for assessing and treating aggression and disruptive behavior was demonstrated by using the naturally occurring reinforces identified in the RAMS to develop treatments that were experimentally tested. The RAMS consists of a series of 2 minute components in which a naturally occurring consequence is applied contingent on the target's behavior, alternated with components in which the consequence is not applied, with no break between components. The aggressive and disruptive behaviors of four 2- to 11-year-old children were analyzed and treated in school and home settings. The RAMS analyses yielded clear results about the reinforcing function of naturally occurring consequences in all cases, and the treatments using the reinforcers identified in the RAMS were all effective. The possible uses of the RAMS as an efficient, ecologically and experimentally valid tool for clinical assessment are discussed. PMID- 10467890 TI - Relaxation to assist sleep medication withdrawal. AB - This study explores the usefulness of relaxation and gradual medication withdrawal in weaning insomniacs from sleep (hypnotic) medication. We recruited 40 volunteers from the community who had insomnia, half of whom were chronic users of hypnotics while the other half were nonmedicated. Half of all participants (10 medicated and 10 nonmedicated) received progressive relaxation. All medicated participants received a standard gradual drug withdrawal program. Medicated participants reduced sleep medication consumption by nearly 80%. Participants who received relaxation obtained additional benefits in sleep efficiency, rated quality of sleep, and reduced withdrawal symptoms. Medicated and nonmedicated participants attained comparable, improved sleep by posttreatment and follow-up. Hypnotic withdrawal was accompanied by serious worsening of insomnia, but this dissipated by the end of the withdrawal period. The psychological treatment of hypnotic-dependent insomnia has high potential for making an important clinical contribution. PMID- 10467891 TI - Promoting generalized social interactions using puppets and script training in an integrated preschool. A single-case study using multiple baseline design. AB - Puppet script training was used to teach the social skills of greeting, responding to conversations, and initiating conversations to a preschool child with visual impairments. Susie and four peers without disabilities were taught social skills utilizing puppets enacting sociodramatic scripts within group training sessions. Training sessions were immediately followed by free-play activities among peers without disabilities to assess skill generalization. A single-case study using a multiple-baseline design demonstrated that the intervention increased performance of social skills during recess with peers. Results demonstrated that Susie learned the target behaviors and generalized their use to free-play activities with her peers. PMID- 10467892 TI - The modification of breathing behavior. Pavlovian and operant control in emotion and cognition. AB - The purpose of this article is to bring attention to breathing as a behavior that can be modified by means of Pavlovian and operant principles of control. With this aim in mind, this paper (a) reviews a selection of early and recent conditioning studies (Pavlovian and operant paradigms) in respiratory psychophysiology, (b) discusses the bidirectional relationship between breathing and emotion/cognition, and (c) discusses theoretical and applied implications that point to new directions for research in the laboratory and clinic. Emphasis is placed on dyspnea/suffocation fear and the acquisition of anticipatory dyspnea/suffocation fear in panic, anxiety, and stress disorders and their concomitant cognitive deficits. Discussions throughout the article focus on research relevant to theory and application, especially applications in programs of remedial breathing (breathing retraining) designed for the treatment of psychophysiological disorders (e.g., panic, anxiety, and stress) and the accompanying cognitive deficits that result from cerebral hypoxia induced by conditioned hyperventilation. PMID- 10467893 TI - Using self-instructional pictorial manuals to teach child-care skills to mothers with intellectual disabilities. AB - Children of parents with intellectual disabilities (i.e., IQs less than 80, labeled as having mental retardation) are at risk for neglect due to inadequate parenting abilities. Previous studies have shown that these parents are responsive to parent-training packages consisting of instructions, pictorial cues, modeling, feedback, and reinforcement. This study evaluated the effectiveness of self-learning pictorial-parenting manuals in teaching basic child-care skills (diapering, treating diaper rash, bathing, safety) to parents with intellectual disabilities who are monitored by child protection agencies. The manuals alone increased child-care skills (to levels seen in parents without disabilities) in 9 out of the 10 mothers in the study and in 12 of 13 child-care skills. The remaining skill was acquired with the full training package. Follow up indicated that the acquired skills were maintained for up to 3 years. Mean correct performance with the manual was positively correlated with the trainer's rating of the mother's reading level and acceptance of the manual when the mother was first given the manual. Consumer satisfaction ratings of the manuals were high. These results indicate that many parents with intellectual disabilities may improve their child-care skills without intensive training and that self instruction may be an easily disseminable and cost-effective way of reducing the risk of child neglect due to parenting skill deficiencies. PMID- 10467894 TI - Mortality and morbidity of members of the British Nuclear Tests Veterans Association and the New Zealand Nuclear Tests Veterans Association and their families. AB - More than 20,000 British servicemen--many of them on their National Service, few of them volunteering for the tests, and most in their early twenties, some still boy soldiers in their teens--were required to 'participate' in the United Kingdom nuclear tests in Australia and Christmas Island in the 1950s and 1960s. 528 members of the New Zealand Navy were also present for one series of tests. There was also a Fijian Army contingent, which has been variously numbered between 100 and 500 men. An estimated 16,000 Australian servicemen and civilians were also involved in the tests at Maralinga and other sites. The men performed a wide range of duties, from highly technical preparations for the detonations to catering and clerical jobs. But whatever their role, they were all required to witness the detonations as part of their 'indoctrination' for the possibility of nuclear war. Most of them were required to line up on the beach, with their backs to the detonations and their hands over their eyes for the first minute or so. They were then allowed to turn around and look at the awesome sight as the mushroom cloud plumed thousands of feet into the air. Very few wore more than shorts and sandals during their time at the tests; only those who were thought to be at risk from radiation injury were issued with protective clothing and radiation dose badges. The UK government was sure that the troops, most of whom were standing within 20 km of the detonations and some of whom were present for 25 nuclear bomb blasts in as many weeks on Christmas Island, were not irradiated. The Ministry of Defence still routinely issues a document to nuclear veterans who feel that their illnesses were caused by the radiation they encountered when they were young men which states: The background [radiation] dose received by civilians and members of HM Forces serving at or off Christmas Island in the years 1956 to 1964 was only about 35% of that which they would have received on average had they remained, for that period of their lives, in the United Kingdom- that is, some 100 microsieverts per calendar month less at Christmas Island than in the United Kingdom. This sanguine view of the health burden borne by nuclear veterans and their families is not borne out by the data reported in this study of the health outcomes of the 2,500 men (2,200 UK, 238 New Zealand and 62 Fijian) on whom data are available to the present researcher. Thirty per cent of the men in this sample have already died, mostly in their fifties. Two-thirds of them died from cancers that are pensionable in the United States as presumptively radiogenic among nuclear veterans. About one in seven of the men in the sample of 1,014 who responded to the questionnaire circulated in late 1997 did not father any children after they returned from the weapons tests. Among the nearly 5,000 children and grandchildren of this group of more than a thousand veterans, there are 26 cases of spina bifida alone--more than five times the usual rate for live births in the UK. Nearly half the health problems among the offspring of the nuclear weapons tests veterans reported in this study consist of the same dermatological, musculoskeletal and gastrointestinal conditions from which many of the men have also suffered. Among the 2,261 children of 1,041 veterans, more than 200 skeletal abnormalities were reported, including more than 30 cases of short stature and 18 spinal problems, mostly curvature and scoliosis. More than 100 skin conditions were reported, mostly eczema and dermatitis, in many cases described as congenital. Over 50 of the children are already suffering from arthritis and similar conditions, although they are only now entering their thirties. Hip deformities were reported for 19 children and kneecap deformities for 14. More than 100 of the veterans' children reported reproductive difficulties; 24 women reported problems with their ovaries. This pattern of morbidity was repeated in the grandchildren, tho PMID- 10467895 TI - Risk for psychopathology in the children of depressed mothers: a developmental model for understanding mechanisms of transmission. AB - A large body of literature documents the adverse effects of maternal depression on the functioning and development of offspring. Although investigators have identified factors associated with risk for abnormal development and psychopathology in the children, little attention has been paid to the mechanisms explaining the transmission of risk from the mothers to the children. Moreover, no existing model both guides understanding of the various processes' interrelatedness and considers the role of development in explicating the manifestation of risk in the children. This article proposes a developmentally sensitive, integrative model for understanding children's risk in relation to maternal depression. Four mechanisms through which risk might be transmitted are evaluated: (a) heritability of depression; (b) innate dysfunctional neuroregulatory mechanisms; (c) exposure to negative maternal cognitions, behaviors, and affect; and (d) the stressful context of the children's lives. Three factors that might moderate this risk are considered: (a) the father's health and involvement with the child, (b) the course and timing of the mother's depression, and (c) characteristics of the child. Relevant issues are discussed, and promising directions for future research are suggested. PMID- 10467896 TI - Phonology, reading acquisition, and dyslexia: insights from connectionist models. AB - The development of reading skill and bases of developmental dyslexia were explored using connectionist models. Four issues were examined: the acquisition of phonological knowledge prior to reading, how this knowledge facilitates learning to read, phonological and nonphonological bases of dyslexia, and effects of literacy on phonological representation. Compared with simple feedforward networks, representing phonological knowledge in an attractor network yielded improved learning and generalization. Phonological and surface forms of developmental dyslexia, which are usually attributed to impairments in distinct lexical and nonlexical processing "routes," were derived from different types of damage to the network. The results provide a computationally explicit account of many aspects of reading acquisition using connectionist principles. PMID- 10467897 TI - A neuropsychological theory of positive affect and its influence on cognition. AB - Positive affect systematically influences performance on many cognitive tasks. A new neuropsychological theory is proposed that accounts for many of these effects by assuming that positive affect is associated with increased brain dopamine levels. The theory predicts or accounts for influences of positive affect on olfaction, the consolidation of long-term (i.e., episodic) memories, working memory, and creative problem solving. For example, the theory assumes that creative problem solving is improved, in part, because increased dopamine release in the anterior cingulate improves cognitive flexibility and facilitates the selection of cognitive perspective. PMID- 10467898 TI - The primacy of primary control is a human universal: a reply to Gould's (1999) critique of the life-span theory of control. AB - This reply to S. J. Gould's (1999) critique of J. Heckhausen and R. Schulz's (1995) life-span theory of control addresses four issues: (1) the universal claim that primary control holds functional primacy over secondary control, (2) the status of secondary control as a confederate to primary control, (3) empirical evidence and paradigms for investigating universality and cultural variations, and (4) the capacity of the human control system to manage both gains and losses in control throughout the life span and aging-related decline in particular. Theoretical perspectives and empirical evidence from evolutionary, comparative, developmental, and cultural psychology are presented to support the authors' view that primary control striving holds functional primacy throughout the life span and across cultural and historical settings. Recommendations for empirically investigating the variations in the way primary control striving is expressed in different cultures are outlined. PMID- 10467899 TI - Comparison and contrast as a mechanism of perceptual learning? AB - In a series of flavour aversion experiments, rats received different schedules of pre-exposure to two compound flavours (AX and BX). Discrimination between them was assessed by establishing an aversion to AX and measuring generalization of this aversion to BX. Experiment 1 demonstrated that alternating pre-exposure to AX and BX resulted in less generalization than did blocked exposure, where animals received all exposure to AX before exposure to BX (or vice versa). This difference was not accompanied by any difference in the strength of the aversion conditioned to the common X element. Varying the interval between exposure to AX and BX in the alternating condition from a minute or two to several hours had no effect on generalization. However, Experiments 2 and 3 showed that when the interval between exposure to AX and that to BX was reduced to zero sec, the alternating schedule increased generalization between AX and BX. In this case, the increase in generalization was accompanied by an increase in the strength of the aversion conditioned to X. PMID- 10467900 TI - Neuropharmacological evidence for different timing mechanisms in humans. AB - Temporal processing of intervals in the range of seconds or more is cognitively mediated, whereas processing of brief durations below 500 msec appears to be based on brain mechanisms outside cognitive control. To elucidate the critical role of various neurotransmitters in timing processes in humans, the effects of 3 mg of haloperidol, a dopamine receptor antagonist, 11 mg of the benzodiazepine midazolam, and 1 mg of scopolamine, a cholinergic receptor antagonist, were compared in a placebo-controlled double-blind experiment. In addition, changes in cortical arousal, semantic memory, and cognitive and motor skill acquisition were assessed. Temporal processing of long durations was significantly impaired by haloperiodol and midazolam, whereas processing of extremely brief intervals was only affected by haloperidol. The overall pattern of results supports the notion that temporal processing of longer intervals is mediated by working-memory functions and, therefore, any pharmacological treatment, irrespective of the neurotransmitter system involved, that produces a deterioration of working memory, may interfere with temporal processing of longer intervals. Temporal processing of intervals in the range of milliseconds appears to depend on the effective level of dopaminergic activity in the basal ganglia. PMID- 10467901 TI - Development of obstructive hydrocephalus with lumboperitoneal shunting following subarachnoid hemorrhage. AB - Hydrocephalus is a frequent complication of subarachnoid hemorrhage (SAH). The optimum method of treating hydrocephalus in this setting has not been determined. We review our experience with patients developing communicating hydrocephalus secondary to SAH and subsequently treated with lumboperitoneal (LP) shunts. Following hospitalization for the treatment of SAH, patients who developed clinical symptoms and radiologic signs of hydrocephalus were treated with (ventriculoperitoneal) VP or LP shunting. Eighteen patients received an LP shunt, of which seven (28%) developed a non-communicating or obstructive hydrocephalus. These seven patients underwent replacement with a VP shunt and have not had further complications. In the setting of post-SAH communicating hydrocephalus, obstructive hydrocephalus may develop after LP shunt placement. Patients who develop this complication and have their LP shunts converted to VP shunts have a favorable prognosis. PMID- 10467902 TI - Electromyographic findings in shoulder dislocations and fractures of the proximal humerus: comparison with clinical neurological examination. AB - There is no consensus of opinion about the frequency of associated nerve lesions in anterior shoulder dislocations and fractures of the proximal humerus. We undertook a prospective study to assess the incidence, the severity of the nerve injury and the diagnostic value of electromyographic examination; 215 patients were included. We performed neurological examination and needle electromyography (EMG). Nerve injury was graded according to a denervation score at the EMG. EMG disorders were seen in 133 patients (62%). Testing of sensibility and clinical reflexes proved not to be a reliable indicator for EMG abnormalities. Detection of axonal lesions by grading muscle strength based on the MRC score after these shoulder traumas is difficult. The findings of this study imply that by clinical examination alone a large number of axonal lesions remain undetected. PMID- 10467903 TI - Cerebral arteriovenous malformations and movement disorders. AB - A series of six patients with movement disorders associated with cerebral arteriovenous malformations (AVM) is reported. The AVMs were classified according to the Spetzler-Martin classification as grade V (one patient), grade IV (four patients), and as grade III (one patient). One patient had action-induced hemidystonia caused by a contralateral frontoparietal AVM which compressed the putamen and was supplied partially by enlarged lenticulostriate arteries. Two patients presented with unilateral cortical tremor associated with contralateral high-frontal cortical/subcortical AVMs sparing the basal ganglia. Another patient developed hemidystonia and hemichorea-hemiballism after bleeding of a contralateral temporooccipital AVM and subsequent ischemia. Two patients had focal dystonia after thalamic and basal ganglia hemorrhage from AVMs. Five patients were operated on. The movement disorder was abolished in one patient postoperatively. Different mechanisms were identified that are relevant for the development of AVM-related movement disorders: mass effect, diaschisis, local parenchymal altered cerebral blood flow, and hemorrhagic or ischemic structural lesions. PMID- 10467904 TI - Predictive clinical factors of very early in-hospital mortality in subarachnoid hemorrhage. AB - This study was conducted to determine clinical predictors of very early in hospital mortality (within the first 72 h) in patients with non-traumatic subarachnoid hemorrhage. Data of 184 patients with subarachnoid hemorrhage were obtained from consecutive stroke patients included in the prospective Barcelona Stroke Registry. Demographic, anamnestic, clinical, neurological and neuroimaging variables in the subgroup of patients who died within 72 h after the onset of symptoms were compared with those in the subgroup of patients that had survived this initial period. The independent predictive value of each variable on the development of very early death was assessed with a logistic regression analysis. Very early in-hospital death was observed in 18 patients (9.8%). These patients were significantly more likely to have progressive deficit, seizures, altered consciousness, limb weakness, sensory involvement and basal ganglia hematoma than patients without very early death. After multivariate analysis, only progressive deficit (odds ratio (OR) 6.90; 95% confidence interval (95% CI) 2-23.80) and limb weakness (OR 5.46; 95% CI 1.78-16.77) were independent clinical predictors of very early mortality. Progressive neurological deficit and limb weakness at the onset of stroke was independent predictive factors of very early death in patients with non-traumatic subarachnoid hemorrhage. These results further emphasize the need to establish an early etiological diagnosis and to manage these patients aggressively including early surgery in selected cases. PMID- 10467905 TI - A case of Ollier's disease associated with two intracerebral low-grade gliomas. AB - Ollier's disease, or multiple enchondromatosis, is a deforming dysplastic disease of cartilage, characterized by multiple, asymmetrically distributed intra-osseous cartilaginous masses in the metaphyses and diaphyses of bones. When associated with soft tissue hemangiomas it is referred to as Maffucci's syndrome, in which the enchondromatosis has no unilateral distribution. The emergence of malignant neoplasms, including gliomas, is a well-recognized complication in Maffucci's syndrome. We report a 28-year-old patient with a history of Ollier's disease, who developed two low-grade cerebral gliomas as well as an intracranial chondroma. This case history questions the distinction between the two forms of enchondromatosis and supports a continuum between these disease entities. PMID- 10467906 TI - Metastatic adenocarcinoma masquerading as basal pontine tuberculoma. AB - Tuberculous infection of the central nervous system is common in Hong Kong. A 39 year-old woman presented with isolated right sixth nerve palsy which was non progressive for 10 months. Neuro-imaging revealed a right pontine lesion. Cerebrospinal fluid (CSF) examination showed lymphocytic meningitis with negative bacteriological and cytological studies. Empirical antituberculous drugs with initial corticosteroid resulted in improved CSF parameters. A diagnosis of cerebral tuberculoma complicated by meningitis was made. She subsequently deteriorated clinically and radiologically. Despite a number of clinical features which were atypical of leptomeningeal metastasis, adenosquamous carcinoma was found on biopsy. Her relatively indolent clinical course might be due to the initial corticosteroid treatment. This report illustrates the importance of early tissue diagnosis in uncertain cases of chronic lymphocytic meningitis. PMID- 10467907 TI - Hyponatremia-induced metabolic encephalopathy caused by Rathke's cleft cyst: a case report. AB - Rathke's cleft cysts are sometimes associated with aseptic meningitis or metabolic encephalopathy due to hyponatremia. We treated such a case manifest by lethargy, fever and electroencephalographic abnormalities. A 68-year-old man was admitted to our ward after experiencing general malaise, nausea and vomiting and then high fever and lethargy. On admission, he was drowsy and had nuchal rigidity and Kernig's sign. Physically, he was pale with dry, thickened skin. He had lost 5.0 kg of body weight in the last month. His serum sodium was 115 mEq/l. He had a low serum osmotic pressure (235 mOsmol/l) and a high urine osmotic pressure (520 mOsmol/l). His urine volume was 1200-1900 ml/24 h with a specific gravity of 1008 1015. The urine sodium was 210 mEq/l. He did not have an elevated level of antidiuretic hormone. Electroencephalograms showed periodic delta waves over a background of theta waves. With sodium replacement, the patient become alert and symptom free, and his electroencephalographic findings normalized. However, the serum sodium level did not stabilize, sometimes falling with a recurrence of symptoms. Magnetic resonance imaging clearly delineated a dumbbell-shaped intrasellar and suprasellar cyst. The suprasellar component subsequently shrunk spontaneously and finally disappeared. An endocrinologic evaluation showed panhypopituitarism. The patient was given glucocorticoid and thyroxine replacement therapy, which stabilized his serum sodium level and permanently relieved his symptoms. A transsphenoidal approach was performed. A greenish cyst was punctured, and a yellow fluid was aspirated. The cyst proved to be simple or cubic stratified epithelium, and a diagnosis of Rathke's cleft cyst was made. The patient was discharged in good condition with a continuation of hormonal therapy. Rathke's cleft cyst can cause aseptic meningitis if the cyst ruptures and its contents spill into the subarachnoid space. Metabolic encephalopathy induced by hyponatremia due to salt wasting also can occur if the lesion injures the hypothalamus and pituitary gland. PMID- 10467908 TI - Wernicke's encephalopathy associated with hemodialysis: report of two cases and review of the literature. AB - Two rare cases of Wernicke's encephalopathy (WE) in non-alcoholic patients on hemodialysis (HD) are reported. They presented with the clinical triad of WE (ophthalmoplegia, ataxia and disturbance of consciousness) and intravenous administration of thiamine led to complete elimination of these manifestations. Reduced plasma thiamine levels prior to the administration confirmed the diagnosis of WE. Interestingly, a reduction in plasma thiamine levels by about half was seen in one of the patients on HD, suggesting that thiamine, a water soluble vitamin, can be depleted with HD. In the literature, nine HD-dependent patients have been reported to develop WE, seven of whom were diagnosed postmortem. Their premortem diagnoses included uremic encephalopathy, dysequilibrium syndrome and dialysis dementia, which can often complicate HD and present symptoms similar to those of WE. We therefore emphasize that WE, even though a rare complication, should be suspected in all patients on HD who present with at least one of the clinical triad of WE. PMID- 10467909 TI - Magnetic resonance imaging findings of Kernohan-Woltman notch in acute subdural hematoma. AB - OBJECTIVE AND IMPORTANCE: We report the case of a 73-year-old patient who presented a right motor deficit caused by an ipsilateral acute subdural hematoma. A magnetic resonance imaging (MRI) demonstration of Kernohan-Woltman notch phenomenon was obtained. CLINICAL PRESENTATION: The woman sustained a major head injury at home, followed by loss of consciousness. On admission to the emergency room, she was comatose, anisochoric (left > right), and showed a reaction to pain with decerebrating movements of left limbs (Glasgow Coma Scale (GCS) 4/15). A right severe hemiparesis was observed. Cerebral computed tomography scan showed a large right hemispheric subdural hematoma. INTERVENTION AND POST-OPERATIVE COURSE: A wide right craniotomy was performed and the subdural hematoma evacuated. During the post-operative period, the level of consciousness gradually improved. A MRI performed about 2 weeks after operation showed a small area of abnormal signal intensity in the left cerebral peduncle. On discharge, the woman was able to communicate with others, but her right hemiparesis was still severe. PMID- 10467910 TI - Efficacy of spinal cord stimulation for neuropathic pain following idiopathic acute transverse myelitis: a case report. AB - Spinal cord stimulation (SCS) involves the electrical stimulation of dorsal structures within the spinal cord, and is the most widespread application of neurostimulation for the relief of chronic pain. Idiopathic acute transverse myelitis (IATM) is an acute monophasic lesion of the spinal cord that presents with symptoms associated with loss of cord function. The incidence of chronic pain secondary to this condition is unknown, but is considered rare. We report the efficacy of SCS for relief of severe neuropathic pain in both lower limbs secondary to IATM, which had failed to respond to conventional pain therapies. PMID- 10467911 TI - Pituitary adenoma with neuronal choristoma: a report of two rare cases. AB - Two rare cases of pituitary adenoma with neuronal choristoma are described. Both patients were female and presented with features of acromegaly with elevated growth hormone and prolactin levels. Radiologically, both lesions were predominantly intrasellar in location with extension into suprasellar region, but hypothalamus was not involved. Histopathological examination revealed a mixture of chromophobe pituitary adenoma cells and neuronal cells. In both cases, the adenoma component was positive for growth hormone and prolactin. Interestingly, immunopositivity for alpha-subunit, cytokeratin and prolactin was seen in the adenoma and neuronal cells in one case. Our findings support the hypothesis that the neuronal cells possibly arise from adenoma cells as a result of metaplasia. PMID- 10467912 TI - Fluctuating clinical myotonia and weakness from Thomsen's disease occurring only during pregnancies. AB - Advances in molecular genetics are allowing better phenotype to genotype correlation of the non-dystrophic myotonic disorders. We report a 32-year-old woman, who first noted myotonia that was associated with weakness during her first pregnancy. The work-up disclosed that she had Thomsen's disease which is not known to be associated with weakness. In addition, her myotonia was of the fluctuating type and occurred (symptomatically) only during two pregnancies. We discuss the evaluation of myotonia in the pregnant woman which led to the diagnosis of Thomsen's disease and we conclude that in exceptional cases, fluctuating myotonia and weakness occurs in autosomal dominant chloride channel myotonia (Thomsen's disease). PMID- 10467913 TI - X-linked adrenoleukodystrophy: spinocerebellar variant. AB - The phenotypic variability in X-linked adrenoleukodystrophy (X-ALD) can be wide and varied. Rarely, it can present with clinical signs of spinocerebellar degeneration. There are very few reported cases of selective predominant white matter disease of the cerebellum in these patients. We report a patient with a rare variant of adult onset ALD who was previously diagnosed as spinocerebellar ataxia. He was a 24-year-old male who had delayed developmental milestones, developed signs of spinocerebellar degeneration (SCD) after 10 years of Addison's disease. Serial Magnetic Resonance Imaging (MRI), revealed cerebellar and pontine white matter disease but sparing the cerebral cortex and supratentorial white matter. His diagnosis of X-ALD was subsequently confirmed by the elevated serum very long chain fatty acids. This patient illustrates the unusual clinical presentation and imaging features of X-ALD and the importance of considering X ALD in the clinical context of spinocerebellar degeneration. Early recognition of this rare variant would allow proper genetic counselling and institution of dietary therapy and/or bone marrow transplantation. PMID- 10467914 TI - Expanding cyst following temporal lobectomy: an unusual complication of epilepsy surgery. AB - Following anterior temporal lobectomy performed to control intractable complex partial seizures (CPS), it is rare to find a symptomatic cyst at the lobectomy site causing increased intracranial pressure and neurological deterioration. We report a 24-year-old lady who underwent anterior temporal lobectomy with extended amygdalohippocampectomy for CPS of temporal lobe origin. Ten months following the procedure, she developed a large expanding cyst at the temporal lobectomy site manifesting with recurrence of CPS, progressive focal neurological deficit and increased intracranial pressure. The patient underwent a repeat craniotomy, decompression of the cyst along with wide excision of the wall and fenestration of the arachnoid membrane into the basal cisterns. Following the procedure, the features of increased intracranial pressure and focal neurological deficit promptly improved and her seizures became better controlled. Craniotomy and fenestration of a symptomatic iatrogenic cyst following temporal lobectomy results in clinical improvement, obviating the need for a permanent cystoperitoneal shunt. PMID- 10467915 TI - Laminar cortical necrosis in central nervous system lupus: sequential changes in MR images. AB - A 44-year-old woman with systemic lupus erythematosus developed central nervous system lupus presenting with headache, fever, cloudiness of consciousness, and psychotic symptoms. T1-weighted and proton MR images showed laminar high intensity lesions in the parietal and temporal cortex bilaterally. T2-weighted images of the lesions showed low signal intensity. Treatment with corticosteroids alleviated the clinical symptoms within 7 months. The low T2 and high T1 signal abnormalities disappeared in a year and in 5 years, respectively. The laminar cortical lesions on MR images were suggested to represent cortical necrosis associated with central nervous system lupus. PMID- 10467916 TI - Symptomatic sacral extradural arachnoid cyst associated with lumbar intradural arachnoid cyst. AB - A case of sacral extradural arachnoid cyst associated with lumbar intradural arachnoid cyst in a 35-year-old male is reported. The patient presented with a history of severe sacrococcygeal pain, constipation, and dysuria for several months. Computed tomographic (CT) myelograms and magnetic resonance imaging (MRI) scans showed a huge sacral cyst without neural components. A favorable outcome could be achieved by decompression of the cyst, obliteration of the fistulous channel between the cyst and the thecal sac, and fenestration of the arachnoid cyst into the subarachnoid space. The relevant literatures are also reviewed. PMID- 10467917 TI - Use and efficacy of low-dose ketamine in the management of acute postoperative pain: a review of current techniques and outcomes. AB - Ketamine hydrochloride is a well known general anesthetic and short acting analgesic in use for almost 3 decades. The role of the NMDA receptor in the processing of nociceptive input has led naturally to renewed clinical interest in N-methyl-D-aspartate (NMDA) receptor antagonists such as ketamine. This paper reviews the use and efficacy of low-dose ketamine in the management of acute postoperative pain. The literature was obtained from a computer search of the MEDLINE database from 1966 through December 1998. Studies were included for review if they were randomized, prospective, controlled, double-blind and reported pain scores. We evaluate the clinical literature and discuss the efficacy of low-dose ketamine in the management of acute postoperative pain when administered alone or in conjunction with other agents via the oral, intramuscular, subcutaneous, intravenous and intraspinal routes. Low-dose ketamine is defined as a bolus dose of less than 2 mg/g when given intramuscularly or less than 1 mg/kg when administered via the intravenous or epidural route. For continuous i.v. administration low-dose ketamine is defined as a rate of < or =20 microg/kg per min. We conclude that ketamine may provide clinicians with a tool to improve postoperative pain management and to reduce opioid related adverse effects. The evidence suggests that low-dose ketamine may play an important role in postoperative pain management when used as an adjunct to local anesthetics, opioids, or other analgesic agents. Further research is required in the following areas: (a) dose-finding studies for ketamine as an adjunct to opioids and local anesthetics (b) efficacy and optimal route of administration (c) the role of S(+)-ketamine; (d) the influence of ketamine on long-term outcome such as chronic pain (e) long-term physical and chemical stability of mixtures containing ketamine (f) spinal toxicity of ketamine and (g) effects of low-dose ketamine on cognitive and memory functioning after surgery. PMID- 10467918 TI - C- and A delta-fiber components of heat-evoked cerebral potentials in healthy human subjects. AB - Feedback-controlled laser heat was used to stimulate the hairy skin of the hand dorsum and forearm, and heat-evoked cerebral potentials were recorded at midline (Fz, Cz, Pz) and temporal (T3, T4) scalp positions. Based on data from primary afferent electrophysiology a stimulus level (40 degrees C) was chosen, which is above C-fiber heat threshold, but clearly below A delta-nociceptor heat threshold in order to excite selectively C-fibers without concomitant excitation of A delta fibers. Feedback-controlled stepped heat stimuli to 40 degrees C elicited ultralate laser evoked potentials (LEPs) at the vertex in a high proportion of experiments (90%). Estimates of conduction velocity calculated from latency shifts between the hand and forearm sites of ultralate LEPs (2.4 m/s) and of reaction times (2.8 m/s) confirmed mediation of ultralate potentials by unmyelinated nerve fibers (nociceptors and/or warm fibers). The ultralate LEP could be differentiated from resolution of contingent negative variation (CNV), an endogenous potential related to expectation and response preparation, by its scalp topography. Strong heat stimuli of 48 degrees C, which is suprathreshold for most A delta- and C-fiber nociceptors, elicited the well-known late LEPs mediated by nociceptive Adelta-fibers confirming previous studies. The LEP waveform to strong heat stimuli also contained an ultralate component reminiscent of an ultralate LEP following the late LEP. Ultralate and late LEP had identical scalp topography. In conclusion, the method of temperature-controlled laser heat stimuli allows the selective and reliable examination of A delta- and C-fiber mediated afferent pathways and the related cortical processing without the complication of dissociating A-fiber nerve blocks. PMID- 10467919 TI - Does sex make a difference in the prescription of treatments and the adaptation to chronic pain by cancer and non-cancer patients? AB - The literature suggests that the sex of patients is an important factor in understanding how they are treated by health care professionals and how they adapt to their symptoms. In two groups of patients with chronic pain (n = 428 non cancer (Study 1) and n = 143 cancer-related (Study 2)), men and women were compared on medications prescribed, treatment history, and coping and adaptation. In Study 1 with the non-cancer pain patients, there were no significant differences between the sexes in past treatments, current analgesic use, pain, or disability. Women were significantly more depressed and were more likely to receive antidepressants than men. Subgrouping patients on the basis of pain adaptation responses yielded groups with distinct psychosocial and behavioral characteristics. In Study 2 with the cancer pain patients, men and women did not show significant differences on any variables. Consistent with the results of Study 1, however, psychological subgroups differed significantly in pain severity, mood and disability regardless of sex. The results of both studies suggest that the role of patients' sex in chronic pain may be less important than their psychosocial and behavioral responses. Thus, it appears that knowing the psychological characteristics of patients may be more important than their sex. PMID- 10467920 TI - Peripheral antinociceptive actions of desipramine and fluoxetine in an inflammatory and neuropathic pain test in the rat. AB - Amitriptyline, a non-selective noradrenaline (NA) and 5-hydroxytryptamine (5-HT) reuptake inhibitor, has recently been demonstrated to produce a peripheral antinociceptive action in an inflammatory (formalin test) and a neuropathic pain model (spinal nerve ligation). In the present study, we determined whether desipramine, a selective NA reuptake inhibitor, and fluoxetine, a selective 5-HT reuptake inhibitor, could produce peripheral antinociceptive actions in these same tests. Effects on paw volume also were determined. In the 2.5% formalin test, desipramine and fluoxetine 10-300 nmol produced a dose-related reduction in phase 2 (16-60 min) flinching and biting/licking behaviours when coadministered with the formalin. Phase 1 flinch behaviours (0-12 min) were significantly reduced at the highest dose. These actions are peripherally mediated, as they were not seen when desipramine or fluoxetine (100, 300 nmol) were injected into the contralateral hindpaw. The peripheral action of desipramine and fluoxetine was not altered by coadministration of caffeine 1500 nmol. In the spinal nerve ligation model, desipramine 100 nmol, but not fluoxetine 100 nmol, produced a peripheral anti-hyperalgesic action in the hindpaw corresponding to the ligated side when thresholds were determined using a thermal paw stimulator. In paw volume experiments, desipramine, at doses which are maximally effective in behavioural tests, produced only a slight increase in paw volume, but fluoxetine (10-300 nmol) produced a robust and sustained dose-related increase in paw volume. Amitriptyline also produced minimal effects on paw volume. When coinjected with formalin, no agent significantly altered the degree of paw swelling produced by formalin. The increase in paw volume produced by fluoxetine was inhibited by ketanserin (5-HT2 receptor antagonist), mepyramine (histamine H1 receptor antagonist) and phentolamine (alpha-adrenergic receptor antagonist), but not by the other selective 5-HT receptor antagonists tested or caffeine. The pronounced peripheral pain alleviating actions in the absence of marked changes in paw volume produced by desipramine and amitriptyline, but not fluoxetine, in the formalin test and the spinal nerve ligation model suggest that these agents could be developed as cream or gel formulations to recruit a peripheral antinociceptive action in inflammatory and neuropathic pain states. Such a formulation might permit the attainment of higher and more efficacious concentrations in the region of the sensory nerve terminal, with limited systemic side effects. PMID- 10467921 TI - Dissociation of sensory and affective dimensions of pain using hypnotic modulation. AB - Understanding the complex nature of pain perception requires the ability to separately analyze its psychological dimensions and their interaction, and relate them to specific variables and responses. The present study, therefore, attempted to selectively modulate the sensory and affective dimensions of pain, using a cognitive intervention, and to assess the possible relationship between these psychological dimensions of pain and changes in physiological responses to the noxious stimuli. In three experiments, normal subjects trained in hypnosis rated pain intensity and pain unpleasantness produced by a tonic heat-pain stimulus (1 min immersion of the hand in 45.0-47.5 degrees C water). Two experiments were designed to test hypnotic suggestions to decrease (Experiment one (Section 2.5.1)), or increase and decrease (Experiment two (Section 2.5.2)) pain affect. Suggestions in Experiment three (Section 2.5.3) were directed towards an increase or decrease in pain sensation. In Experiments one and two (Sections 2.5.1 and 2.5.2), the significant modulation in pain unpleasantness ratings was largely independent of variations in perceived pain intensity. Moreover, in Experiment two (Section 2.5.2), there was a significant correlation between the stimulus evoked heart-rate increase and ratings of pain unpleasantness, but not of pain intensity, suggesting a direct functional interaction between pain affect and autonomic activation. In Experiment three (Section 2.5.3), suggestions to modulate the sensory aspect of pain produced significant modulation of pain intensity ratings, with secondary changes in pain unpleasantness ratings. Hypnotic susceptibility (Stanford Hypnotic Susceptibility Scale form A) was specifically correlated to pain unpleasantness modulation in Experiment two (Section 2.5.2) and to pain intensity modulation in Experiment three (Section 2.5.3), suggesting that this factor relates to the primary process toward which hypnotic suggestions are directed. The specific pain dimension on which hypnotic suggestions act depends on the content of the instructions and is not a characteristic of hypnosis itself. Results are consistent with a successive-stage model of pain perception (e.g. Wade JB, Dougherty LM, Archer CR, Price DD. Assessing the stages of pain processing: a multivariate analytical approach. Pain 1996;68:157-167) which provides a conceptual framework necessary to study the cerebral representation of pain perception. PMID- 10467922 TI - NSAID-induced cyclooxygenase inhibition differentially depresses long-lasting versus brief synaptically-elicited responses of rat spinal dorsal horn neurons in vivo. AB - This electrophysiological study examined the effects of NSAID administration on synaptically-elicited responses of rat single spinal dorsal horn neurons to natural stimulation of peripheral receptive fields. Nociceptive responses consisted of a fast initial discharge during the stimulus followed by a slowly decaying afterdischarge. The cyclooxygenase inhibitor, indomethacin (2.0-8.0 mg/kg, i.v.), was without effect on the on-going rate of discharge but dose dependently inhibited synaptically-elicited responses to noxious cutaneous mechanical stimulation (fast initial discharge: n = 3/3 with 2 mg/kg, 5/8 with 4 mg/kg, 5/6 with 8 mg/kg; slowly-decaying afterdischarge: n = 3/3 with 2 mg/kg, 6/8 with 4 mg/kg, 6/6 with 8 mg/kg) and thermal (fast initial discharge: n = 7/9 with 8 mg/kg; slowly-decaying afterdischarge: n = 3/4 with 4 mg/kg, n = 7/9 with 8 mg/kg). The inhibitory effect of indomethacin started within 2-4 min and lasted up to 120 min. To eliminate any effect of indomethacin via cutaneous sensory receptors it was tested on the responses of some neurons to high intensity electrical stimulation of the sciatic nerve; indomethacin depressed these evoked responses (fast initial discharge: n = 5/6 with 2 mg/kg, n = 7/7 with 4 mg/kg; slowly-decaying afterdischarge: n = 6/6 with 2 mg/kg, n = 7/7 with 4 mg/kg). The brief excitatory responses to innocuous pressure (fast initial discharge: n = 2/3 with 2 mg/kg, n = 6/8 with 4 mg/kg, n = 4/6 with 8 mg/kg) and hair (n = 2/7 with 2 and 4 mg/kg, respectively) stimulation in both non-nociceptive and wide dynamic range neurons were also depressed but to a lesser extent. However, the prolonged excitation of three wide dynamic range neurons to continuous hair stimulation was almost entirely inhibited by indomethacin. Overall, inhibition of the afterdischarge and the excitatory effect of long-lasting synaptic input were greater than inhibition of the fast synaptic input-evoked initial discharge. The evidence supports the suggestion that systemically-administered indomethacin has an effect in the spinal cord and demonstrates an action specifically in the dorsal horn. The data are interpreted to suggest that sensory inputs are more involved than input-independent excitation of dorsal horn neurons in leading to de novo synthesis of eicosanoids and that the time course of this synthesis brings the levels to a point where COX inhibition can have an observable effect during prolonged excitation. Although the data suggest that COX inhibition differentially inhibits nociceptive versus non-nociceptive mechanisms at the cellular level, irrespective of the modality of the stimulus, this is the first direct demonstration that prolonged activation of synaptic mechanisms are preferentially inhibited. According to this it would be predictable that NSAIDs would be more effective on nociceptive types of pain characterized by time or prolonged inputs of primary afferents. PMID- 10467923 TI - Estrous variations in behavioral responses to vaginal and uterine distention in the rat. AB - Response properties of peripheral afferent fibers supplying the vagina and uterus of the rat vary with estrous stage (Robbins A, Berkley KJ, Sato Y. Estrous cycle variation of afferent fibers supplying reproductive organs in the female rat. Brain Res 1992,596:353-356), suggesting that behavioral sensitivity to vaginal and uterine stimulation might similarly vary. To test this hypothesis, detection and escape responses of 12 rats to different volumes of distention of the vaginal canal or uterine horn (six rats each) were compared during each of the four estrous stages, proestrus (P), estrus (E), metestrus (M), and diestrus (D), using previously-published behavioral techniques (Berkley KJ, Wood E, Scofield SL, Little M. Behavioral responses to uterine or vaginal distention in the rat. Pain 1995;61:121-131). Estrous variations in vaginal or uterine tone (pressure-volume functions) were also measured in the same rats. Vaginal canal: escape response percentages increased significantly as vaginal distention volume increased, but neither escape nor detection responses varied with estrous stage. Vaginal tone, however, was greater in P and E than in M and D, with the greatest tone in E and the least in D. When escape responses to different pressures were analyzed, it was found that escape response percentages to the same vaginal pressure were lower during P and E than during M and D. One outcome of these estrous influences would be that a vaginal stimulus of a given volume (such as an erect penis) would exert higher pressures during P and E (i.e. the penis would be held within the vaginal canal more firmly) than during M and D, but would be less likely during P and E to provoke escape responses. This modification of nociceptive sensitivity has obvious reproductive advantages, because P and E constitute the rat's fertile period. Uterine horn: escape response percentages increased significantly as uterine distention volume increased only during M and D, with no differences between them. Detection responses also occurred only during M and D. Similarly, uterine tone was significantly greater in M and D than in P and E, with the greatest tone occurring during M and the least during P. Although these changes in uterine tone were opposite to those of the vaginal canal, escape response percentages to the same uterine pressures were, in a manner similar to vaginal pressures, lower during P and E than during M and D. The functional significance of these variations for the uterus is unclear, but does suggest that, under pathophysiological conditions, stimulation of the uterine horn, as well as the vaginal canal, would be more likely to provoke escape behaviors in M and D than in P and E. A similar increased sensitivity in rats during M and D to noxious stimulation of other pelvic organs has been observed by others. This situation resembles that in women, for whom many visceral pain conditions are exacerbated during a comparable part of their ovarian cycle, i.e. perimenstrually. PMID- 10467924 TI - Spinal cannabinoids are anti-allodynic in rats with persistent inflammation. AB - Cannabinoid receptor (CB1) agonists strongly inhibit behavioral responses to acute noxious stimuli, but their effects on behavioral responses in persistent pain states are less clear. Here, we examined the effects of intrathecal (i.t.) administration of a CB1 agonist, WIN55,212-2, on mechanical allodynia (decreased withdrawal threshold) produced by injections of complete Freund's adjuvant (CFA) in the plantar surface of the rat hindpaw. We measured mechanical thresholds with calibrated von Frey filaments before and after CFA and used Fos expression as a marker of the activity of spinal cord neurons during inflammation and in response to a CB1 antagonist. One day post CFA-induced injury, mechanical sensitivity was significantly increased in the hindpaw ipsilateral to the CFA injection, as was the number of neurons that express Fos. Intrathecal injection of WIN55,212-2, significantly, reversed the allodynia at doses that had no effect on the mechanical threshold of the contralateral paw of CFA-treated or the withdrawal thresholds in naive animals. This effect was blocked by coadministration of the CB1 antagonist, SR141716A, with WIN55212-2. By itself, SR141716A, had no effect on mechanical thresholds in normal animals. In inflamed animals, SR141716A did not further reduce mechanical thresholds in the inflamed paw, but it significantly enhanced mechanical sensitivity 'contralateral' to the inflammation. Furthermore, i.t. injection of SR141716A increased Fos expression in both normal and inflamed animals, to a different extent in different laminae. In normal animals, the increase was primarily in laminae V-VI and in the ventral horn; in animals with persistent inflammation SR141716A increased the number of Fos neurons in laminae I-II and V-VI. These results demonstrate that WIN55212-2 reverses inflammation-induced allodynia at doses that do not produce analgesia and that SR141716A differentially affects the pattern of Fos expression in the spinal cord, depending on the presence or absence of inflammation. Taken together, these results suggest that the CB1 receptor system is tonically active in the spinal cord under normal conditions and that its activity is increased in response to injury. PMID- 10467926 TI - Vascular endothelial growth factor and diabetic nephropathy. PMID- 10467925 TI - The effect of fentanyl on c-fos expression in the trigeminal brainstem complex produced by pulpal heat stimulation in the ferret. AB - We have previously shown that Fos-like immunoreactivity (Fos-LI) is evoked in the brainstem of ferrets following stimulation of pulpal A delta and C fibers originating from the maxillary canine. This study evaluated the effects of the mu opioid receptor agonist fentanyl on Fos expression evoked by noxious thermal stimulation of the right maxillary and mandibular canines in pentobarbital/chloral hydrate anesthetized adult male ferrets. Pulpal heating evoked Fos expression in two distinct regions of the spinal trigeminal nuclear complex: the transitional region between subnucleus interpolaris and caudalis (Vi/Vc) and within the subnucleus caudalis (Vc). More Fos positive cells were expressed in both regions ipsilateral to the site of stimulation compared with the contralateral side (P < 0.05, ANOVA). Pretreatment with fentanyl significantly and dose-dependently suppressed the number of Fos positive cells in both the Vi/Vc transitional region and Vc (P < 0.05, ANOVA). The suppressive effect of fentanyl on Fos expression was blocked by the intravenous administration of naloxone, an opioid antagonist, indicating a specific opioid receptor effect. In addition, opioid receptor antagonism with naloxone alone enhanced Fos expression in Vi/Vc and Vc in response to heat stimulation. The administration of naloxone without heat stimulation failed to evoke Fos expression in Vi/ Vc and Vc. These findings suggest that the activation of trigeminal Vi/Vc and Vc neurons by noxious dental heat stimulation is controlled by a naloxone sensitive endogenous opioid system as indicated by Fos expression. Collectively, these results suggest that neuronal populations in Vi/Vc and Vc regions may contribute to pain responses to noxious dental stimulation and these responses can be modulated by both endogenous and exogenous opioids. PMID- 10467927 TI - The ethical challenges of drug company entrepreneurism in modern health care. PMID- 10467928 TI - Elimination of meropenem by continuous hemo(dia) filtration: an in vitro one compartment model. AB - Meropenem is a carbapenem antibiotic with a wide spectrum of activity against most gram positive and gram negative bacteria including anaerobes. Dose adjustments are necessary during continuous renal replacement therapies of acute renal failure. This in vitro study was conducted to investigate the influence of different filter materials, surface areas (AN-69 0.6 m2 and 0.9 m2, polysulfone 0.75 m2, polyamide 0.6 m2), and increasing flow rates (from 3.3 - 26.7 ml/min) on the elimination of meropenem in an in vitro continuous hemo(dia)filtration model. Meropenem was measured using HPLC with UV-detection. While the clearance increased proportionally to increasing dialysate flow rates in filters with a surface area of 0.9 m2, a peak clearance was reached in the small filters at flow rates of 10.0 ml/min (polyamide 0.6 m2) and 18.3 ml/min (AN-69 0.6 m2), when tested under the same conditions. This indicated incomplete dialysate saturation due to the diminished time available for meropenem to equilibrate with the dialysate solution. No adsorption to either of the tested membranes was detected. Dosage recommendations derived from clinical studies might be appropriate when different filter materials, but similar operational settings of the continuous replacement therapy, are applied. Reduction of the recommended dose might be necessary, when renal replacement therapies with lower flow rates and/or filters with smaller surface areas are carried out. PMID- 10467929 TI - Severe gastrointestinal bleeding in a uremic patient treated with estrogen progesterone therapy. AB - Gastrointestinal bleeding is a frequent complication in hemodialysis patients; angiodysplasia is a potential cause, with a higher incidence in uremic patients. We describe a case of severe anemia (Hemoglobin up to 3.5 g/dl) secondary to diffuse angiodysplastic lesions in a hemodialysis patient with mixed connective tissue disease. The case is characterised both by the severity of the clinical picture (extension and entity of angiodysplastic lesions, frequency of bleeding episodes) and by the patient's religious faith which made her reject blood transfusions. We underline the efficacy of estrogen-progesterone therapy in view of the modest results obtained with other therapeutic strategies on bleeding. PMID- 10467930 TI - The enabler cannula pump: a novel circulatory support system. AB - BACKGROUND: The enabler circulatory support system is a catheter pump which expels blood from the left or right ventricular cavity and provides pulsatile flow in the ascending aorta or pulmonary artery. It is driven by a bedside installed pulsatile driving console. The device can easily be implanted by a minimal invasive approach, similar to the Hemopump. PURPOSE: To demonstrate the hemodynamic performance of this new intracardiac support system. METHODS: In a series of 9 sheep, hemodynamic evolutions were recorded in various conditions of myocardial contractility (the non-failing, the moderately failing and the severely failing heart). Heart failure was induced by injection of microspheres in the coronary arteries. RESULTS: Introduction of the cannula through the aortic valve was feasible in all cases. Pump flow by the enabler was gradually increased to a maximum of 3.5 L/min. Diastolic (and mean) aortic blood pressure is significantly increased in the non-failing and moderately failing condition (counterpulsation mode). In heart failure, cardiac output is significantly increased by the pump (p < 0.0001). A drop in left atrial pressure (indicating unloading) is achieved in all conditions but reaches significant levels only during heart failure (p=0.0068). CONCLUSIONS: This new circulatory support system contributes to stabilization of the circulation in the presence of cardiac unloading. In heart failure it actually supports the circulation by increasing cardiac output and perfusion pressure. PMID- 10467931 TI - Vagal nerve activity and the high frequency peak of the heart rate variability. AB - For the Quality of life (QOL) of patients with an artificial heart system, monitoring an information of the cardiovascular control system may be important. We have been evaluating the autonomic nervous system for that purpose. Recently, fluctuations in hemodynamic parameters including heart rate variability (HRV) were evaluated by means of spectral analysis and nonlinear mathematical analysis. Respiratory wavers in HRV were thought to reflect ongoing information of the parasympathetic nerve activity. Is it true? In order to confirm this hypothesis, we recorded vagal nerve activity directly in the chronic animal experiments. Six healthy adult goats were anesthetized with Halothene inhalation and thoracotomy were performed by the fourth lib resection during mechanical ventilation. Arterial blood pressure, right and left atrial pressures were continuously monitored with the catheter insertion. Cardiac output was measured by the electromagnetic flowmeter attached to the ascending aorta. After the chest was closed, incision was made to the left neck and left vagal nerve was separated. Stainless steel electrodes were inserted into the vagal nerve and fixed by the plasticizer. After the incision was closed, the goats were transferred to the cage and extubated after waking. Hemodynamic parameters and vagal nerve activity were measured in the awake condition. The results showed that clear observation of the autonomic nerve discharges were embodied by this experimental system. The vagal nerve discharges were synchronized with heart beat and respiration. The vagal nerve tonus was significantly influenced by the hemodynamic alteration. However in some condition, the respiratory wave was not always consistent with tonus of the vagal nerve activity, thus suggesting that we should check another information to evaluate the parasympathetic tone. We must continue this study to evaluate an autonomic nerve during artificial heart circulation. PMID- 10467932 TI - Effects of anticoagulants on porcine hepatocytes in vitro: implications in the porcine hepatocyte-based bioartificial liver. AB - For the clinical treatment with porcine hepatocyte-based bioartificial liver (BAL), the use of an anticoagulant in the extracorporeal system is essential. In this experiment, we studied the effect of various anticoagulants on cultured porcine hepatocytes. Porcine hepatocytes were isolated and seeded at a density of 2 x 10(5) cells on a collagen-coated plate in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal calf serum (FCS). Twenty-four hours later, the medium was changed to DMEM with various anticoagulants such as nafamostat mesilate (NM), sodium heparin (SH) and sodium citrate (SC) at concentration used clinically. As a control, the hepatocytes were cultured in only DMEM. After culturing for 6 hours, the viability of the porcine hepatocytes, lactate dehydrogenase (LDH) release, lidocaine clearance (cytochrome p450 function) and albumin synthesis were investigated. SC did not affect either the viability or the p450 function of the hepatocytes. In the NM group, the viability of porcine hepatocytes and lidocaine clearance were decreased significantly more than in the other groups. SH did not affect the viability of porcine hepatocytes, however, it seemed to reduce the p450 function. In conclusion, SC may therefore be the optimal anticoagulant available for hepatocyte-based BAL circuit in terms of its cell toxicity. PMID- 10467933 TI - Large volume leukapheresis for collecting hemopoietic progenitors: role of CD 34+ precount in predicting successful collection. AB - In this work we evaluated the efficacy of stem cell collection with Large Volume Procedures. (LVP), and analysed the importance of the CD34+ cell precount in promoting the collection of a sufficient number of CD34+ cells for transplantation, using the Univariate Logistic Regression analysis. Eighty-nine leukapheresis were performed in 49 patients with hematological malignancies and solid tumors, mobilized with chemotherapy plus Granulocyte Colony Stimulating Factor (G-CSF). For each procedure 15.8 liters of blood were processed. The median value of Nucleated Cells (NC) and CD34+ cells precount was respectively 8.29 x 10(9)/ml (range 1.13/45.4) and 43.08 x 103/ml (range 1.06/795.2). Results show the capability of LVP to collect large quantities of hemopoietic progenitors with a median CD34+ cell total yield of 215.02 x 10(6) (range 5.03/2210). The yields per patients' body weight were: CD34+ cells 3.23 x 10(6)/kg (range 0.081/41.58). The regression analysis between blood cell precounts and collection yields gave the following correlations: the CD34+ cell precount correlates with CD34+ yield (r = 0.78 p < 0.00) and with CD34+ cell yield/kg (r = 0.76 p < 0.00). The number of CD34+ cells processed correlated with the number of CD34+ cells collected/kg (r = 0.83 p < 0.000). To investigate the importance of CD 34+ cell precount in promoting CD34+ cell yields > or =2.5 x 10(6)/kg we performed a Univariate Logistic Regression analysis that showed in our patients a probability of collecting > or =2.5 x 10(6) CD34+/kg that rose from 0.6 to 0.95 for CD 34+ precounts that oscillated from 30 to 40 x 10(3) CD34+ cells/ml, respectively. The Univariate Logistic Regression gave a probability of collecting > or =2.5 x 10(6) CD34+ cells/kg that oscillated between 0.64/0.98 for values of CD34+ cells processed from 6 x 10(6)/kg to 8 x 10(6)/kg, p < 0.000. Sixty-three percent of patients reached the target dose of 2.5 x 10(6) CD34+ cells/kg with only one LVP. Until now 12 patients have been transplanted and all have had a prompt and complete lasting recovery. These results confirm the efficacy of LVP in harvesting hemopoietic progenitors and their ability in reconstituting hemopoiesis of transplanted patients, enabling the estimation of CD34+ precounts and CD34+ cells processed values, highly predictive for the collection of > or =2.5 x 10(6) CD34+ cells/kg. Furthermore, the Logistic Model suggests that the best strategy to plan a successful CD34+ cell collection procedure is to identify for each patient the amount of CD34+ cells/kg to be processed rather than the fixed processing of 3/5 blood volumes in all patients. PMID- 10467934 TI - Extracorporeal therapies: should we use plasma instead of blood? AB - The evolution of extracorporeal therapies has required the parallel evolution of devices designed to extract the blood from the patient and to move it in the extracorporeal circuit. Blood is a peculiar fluid in which viscosity may be strongly affected by the contingent operational conditions of the extracorporeal circuit. Because hematocrit is an important component of blood viscosity, and viscosity may definitely affect the performance of blood purification devices, major problems arise when high hematocrits are present in the patient. All these problems might be avoided if plasma could be used instead of whole blood. Plasma treatment is already performed in various therapies, but the treated plasma is obtained by separation from whole blood in the extracorporeal environment. In this paper we propose a new approach to perform different plasma treatments. Plasma is extracted directly from the patient thanks to a special catheter or plasma extraction device. Utilizing plasmafiltration technology to create a membrane that is placed on the tip of an intravascular catheter, plasma extraction may become easy to perform. The extracted plasma is then available for any type of treatment before being restituted to the patient via a second lumen in the same catheter. PMID- 10467935 TI - Adequacy of hemodialysis and cause-specific morbidity. PMID- 10467936 TI - Dextran sulfate-cellulose adsorption in the management of a myeloma patient with renal amyloidosis. PMID- 10467937 TI - Chemokine gene expression in human oral mucosa. AB - In order to gain further understanding of the role of chemokines in healthy oral mucosa, we analyzed mRNA expression of the alpha (CXC)-family chemokines IL-8 and GROgamma as well as of the beta (CC)-family chemokines MIP-1alpha, MIP-1beta and MCP-1 in twenty young and healthy subjects with good oral hygiene. Twenty biopsies were taken from clinically healthy oral mucosa before surgical removal of impacted wisdom teeth. In addition, five biopsies from patients presenting with specific oral lesions were studied. RNA was purified, quantitated and utilized as substrate for competitive reverse transcription-polymerase chain reaction (RT-PCR). In healthy tissue, IL-8 and MCP-1 mRNA was constitutively expressed in all biopsies, whereas GROgamma, MIP-1alpha, and MIP-1beta were significantly lower. These findings suggest that IL8 and MCP-1 play a significant role in oral tissue homeostasis. The few samples from pathological conditions encourage exploring diseased tissue in more detail. PMID- 10467938 TI - Recalcitrance of Streptococcus mutans biofilms towards detergent-stimulated detachment. AB - The biofilm mode of growth protects the plaque microorganisms against environmental attacks, such as from antimicrobials or detergents. Detergents have a demonstrated ability to detach initially adhering bacteria from enamel surfaces, but the ability of detergent components to detach plaque bacteria is not always obvious from in vivo experiments and reports on their clinical efficacy are inconsistent. It is likely that antimicrobials or detergents are unable to penetrate the plaque and reach the bacteria that actually link the plaque mass to the substratum surface. Attenuated total reflectance/Fourier transform infrared spectroscopy was used to measure the transport of sodium lauryl sulphate (SLS) through Streptococcus mutans HG 985 biofilms. The transport of SLS to the base of the S. mutans biofilms was not hindered, while moreover an accumulation of SLS near the base of the biofilms was found, suggesting that SLS was adsorbed to biofilm components. X-ray photoelectron spectroscopy confirmed the ability of S. mutans, grown on sucrose supplemented medium, to adsorb SLS, and simultaneously indicated that exposure of cells to SLS might lead to a loss of surface proteins. Furthermore, experiments in a parallel-plate flow chamber demonstrated that initially adhering S. mutans HG 985 could be stimulated to detach by SLS, but that, depending on the growth stage of the biofilm, only maximally 27% of biofilm bacteria could be stimulated to detach by a 4% (w/v) SLS solution. PMID- 10467939 TI - Effects of bovine immune and non-immune whey preparations on the composition and pH response of human dental plaque. AB - Colostral products from non-immunized cows (CP) and cows immunized with mutans streptococci (IP) were used as mouth rinses in a short-term human study. The acidogenic potential of the products was tested and found to be negligible in vivo before application to subsequent rinsing tests. At first, all the participants received a professional tooth cleaning, after which they rinsed with one of the solutions (IP; CP; water) three times per day for 3 d. After each rinsing period, the resting pH and decrease in plaque pH after sucrose challenge were determined, the amount of plaque was estimated, and all available plaque was collected. No significant differences were recorded in the composition or in the amounts of accumulated plaque. The resting pH values of plaques with low "innate" pH were increased after the IP rinsing period. Surprisingly, the lowest pH values after the sucrose challenge were recorded in IP plaques. The number of cultivable facultative flora or total streptococci were not affected by different rinsings, but the relative number of mutans streptococci significantly decreased after the IP rinsing period when compared to the CP period. Thus, the short term rinsing indicates favourable effects of bovine immune whey on human dental plaque. PMID- 10467940 TI - Effect of in situ plaque mineral supplementation on the state of saturation of plaque fluid during sugar-induced acidogenesis. AB - Dental plaque fluid is normally supersaturated with respect to enamel mineral but this may change to a state of undersaturation when plaque pH falls following sugar exposure, placing the adjacent enamel at risk of caries. We have determined the saturation status of the fluid in both resting and fermenting plaque following mineral supplementation. Eleven subjects abstained from oral hygiene and rinsed their mouth 3 times/d for 3 d with a placebo solution or with test solutions designed to enrich plaque with hydroxyapatite or fluorhydroxyapatite. On the morning of day 4, plaque samples were collected before and after exposure to 10% sucrose. Compared to the placebo, use of the test rinses resulted in significantly higher concentrations of Ca, P and F in plaque residue. In plaque fluid, higher post-sucrose Ca2+ free concentrations and saturation levels with respect to enamel mineral and fluorapatite were found after use of the hydroxyapatite rinse compared to the placebo, effects that probably resulted from the release of cell-bound Ca2+ as well as from the dissolution of apatite. Thus, some evidence was obtained that the test mouthrinses can counteract the fall in saturation level found when plaque is exposed briefly to sucrose. Potential long term benefits of the test mouthrinses deserve further study. PMID- 10467941 TI - Catch-up in mandibular growth after short-term dietary protein restriction in rats during the post-weaning period. AB - Catch-up growth has been defined as growth with a velocity above the statistical limits of normality for age during a defined period of time which follows a period of impaired growth. Since no data are available on catch-up in mandibular growth, the present study was designed to estimate the dynamics of the mandibular size after short-term dietary protein restriction in rats during the post-weaning period. Weanling male rats, 22 d of age, were divided into two groups, control (C) and experimental (E). E rats were fed a protein-free diet during the first 10 d; from this time on, they were placed on a 20% protein diet, as were C rats during the entire experimental period, which lasted 70 d. Five rats from both groups were randomly selected every 10 d and sacrificed. Mandibular growth was estimated directly on the right mandible by measuring several dimensions (mandibular area, base length, mandibular height, mandibular length, alveolar length and incisor alveolar process length). Alveolar and incisor alveolar process lengths did not change with age or dietary protein. All other dimensions increased with age and were thus negatively affected by protein restriction. After growth restriction ceased, the rate of increase of all affected dimensions was above normal values and deficits were swiftly eliminated. Since age independent dimensions compose roughly the anterior portion of the mandible, this portion of the bone was not affected by protein restriction. It was, thus, the posterior part of the mandible which stopped growth during the nutritional insult and showed catch-up during nutritional rehabilitation. In summary, the rat mandible has a high potential for catch-up during the post-weaning period, showing the ability to achieve complete catch-up in about 30 d. PMID- 10467942 TI - Apoptosis in murine calvarial bone and suture development. AB - To study the possible role of apoptosis in calvarial bone and suture development, terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) was performed on whole mount and sectioned calvariae from mice aged between E14 and P6. We also analyzed by in situ hybridization the expression of Msx2, Bmp4 and Bmp7 genes, which are known to act in conserved signaling pathways leading to apoptosis. We found TUNEL-positive cells from E16 onwards in the calvarial bones, intervening sutures and fontanelles. TUNEL-positive osteoblasts and preosteoblasts were identified at or close to the osteogenic fronts, areas of intense osteogenic activity, with TUNEL-positive mesenchymal cells located in the midsutural mesenchyme. TUNEL-positive osteoclasts and osteocytes were also observed in a sporadic fashion, as well as TUNEL-positive dural cells. Msx2 was expressed in the sutural mesenchyme and the dura mater. Bmp4 was expressed in the developing bone, underlying dura mater, the osteogenic fronts, and also weakly in the sutural mesenchyme. Bmp7 was detected at the same locations as Bmp4 but with noticeably stronger intensity in the meninges and overlying epidermis. We propose that this apoptosis is part of normal suture development, and is integral to the balance between bone formation and resorption, so that abnormal apoptosis may lead to premature (Craniosynostosis) or delayed (Cleidocranial dysplasia) suture closure. PMID- 10467943 TI - Development and characterization of an SV40 immortalized porcine ameloblast-like cell line. AB - Enamel is secreted as a protein matrix by the ameloblasts. These same cells then control the maturation of the enamel matrix, secreting proteinases that hydrolyze proteins as mineralization progresses, until mature enamel containing less than 1% protein by weight remains. Further understanding of the factors that control ameloblast function and differentiation requires an in vitro cell culture system. In this study, we report immortalization of enamel organ epithelial cells and the selection of a cell line with characteristics of ameloblasts. Porcine enamel organ cells were dissected from unerupted porcine molars, cultured in serum-free medium, and passaged twice. These cells were transfected with an origin-of replication defective SV40 plasmid by calcium phosphate precipitation, and a cell line with mRNA expression characteristic of ameloblasts was cloned. This cell line (PABSo-E) expressed mRNA for amelogenin, matrix metalloproteinase-20 (enamelysin), and enamel matrix serine proteinase 1 (EMSP1), but not ameloblastin. PABSo-E cells have been passaged more than 55 times, while continuing to maintain characteristics of ameloblasts. These cells will be useful for future studies of ameloblast function. PMID- 10467944 TI - Transplantation of human pulpal and gingival fibroblasts attached to synthetic scaffolds. AB - Autologous tissue grafting for the restoration of oral tissues is limited by several factors, including the availability of sufficient donor tissue. One solution to this problem may be to develop substitute tissue grafts by attaching disaggregated autologous cells propagated in vitro to scaffolds composed of natural or synthetic polymers. We have earlier demonstrated that human dental pulp and gingival fibroblasts (HPF, HGF) adhere to non-woven polyglycolic acid (PGA) scaffolds, proliferate and produce extracellular matrix in vitro. We now report that such HPF and HGF adhered to PGA scaffolds survive when implanted into subcutaneous sites in immuno-compromised mice. The transplanted cells synthesize and secrete type I collagen, cellular fibronectin and may express genes implicated in transducing bone morphogenetic protein (BMP) signals. Messenger RNA for BMP-2, -4, -7 (OP-1), the BMP type I receptors Act RI, BMPR-1A and 1B, the type II receptor BMPR-II, and type I collagen were detected by reverse transcription-polymerase chain reaction (RT-PCR). These data revealed that three adult human dental pulp and gingival cell populations, each from individual donors, attached to PGA scaffolds and cultured for 24 h in vitro, survive implantation and express genes indicative of a capacity to produce extracellular matrix. The implanted cells may also express genes associated with responsiveness to BMP-mediated tissue inductive signals. PMID- 10467945 TI - Antibacterial activity of resin composites with silver-containing materials. AB - Resin composites with antibacterial activity may be useful to decrease the frequency of secondary caries around restorations. The purposes of this study were to investigate the antibacterial activity of light-activated resin composites incorporating one of three silver-containing materials and to evaluate their long-term inhibitory effect against Streptococcus mutans The three types of silver-containing materials, Novaron (N), Amenitop (AM), and AIS, were incorporated into TEGDMA-UDMA-based light-activated resin composites, and the antibacterial activities, mechanical properties and release of silver ions were examined. Minimum inhibitory concentrations in suspensions of N, AM, and AIS against S. mutans were 1.1, 1.2, and 23.0 mg/ml, respectively. Resin composites incorporating 5 wt% of Novaron (N-5) and 7 wt% of Amenitop (AM-7) inhibited the growth of S. mutans after immersion in water for 3 months, whereas the resin composite incorporating 10 wt% of AIS did not. No significant difference in either compressive or flexural strength was observed between the control and N-5 composites after 1 d and 3 months storage in water. However, for AM-5 composite, there was a significant difference in both strength parameters between the two immersion periods. There was no or extremely little release of silver ions from the N-5 and AM-5 composites after 1 d or 3 months immersion in water. These results indicated that a light-activated resin composite incorporating silver containing materials such as Novaron may be clinically useful due to its long term inhibitory effect against S. mutans and favorable mechanical properties. PMID- 10467946 TI - The influence of Carisolv on enamel and dentine surface topography. AB - The purpose of the present study was to investigate the surface topography of healthy enamel and dentine before and after application of a new chemomechanical system for caries removal, Carisolv. The same surfaces were investigated with respect to the influence of phosphoric acid, plus carious dentine after removal with either Carisolv or burrs. One-hundred freshly extracted teeth were used. Surface topography was measured in two different ways in order to characterize the surfaces at different levels of resolution, atomic force microscopy (AFM) and a contact stylus profilometer. Somewhat conflicting data were obtained with the two measuring techniques. When surfaces were investigated over a small area (AFM), healthy enamel seemed unaffected by Carisolv, while healthy dentine became smoother. Etching enamel with phosphoric acid resulted in a rougher surface, while no effect was detected on etched healthy dentine. Caries removal using Carisolv resulted in a smoother surface compared with conventional caries removal. When the surfaces were monitored with the contact profilometer, no effect of Carisolv could be detected on healthy enamel or dentine. Phosphoric acid etching, in contrast, increased the surface roughness of both enamel and dentine. When compared with conventional caries removal technique, caries removal with Carisolv did increase the surface roughness. PMID- 10467947 TI - Numerical simulation of porosity-free titanium dental castings. AB - The objective of this research was to analyse, predict and control the porosity in titanium dental castings by the use of numerical simulation. A commercial software package (MAGMASOFT) was used. In the first part of the study, a model casting (two simplified tooth crowns connected by a connector bar) was simulated to analyse shrinkage porosity. Secondly, gas pores were numerically examined by means of a ball specimen with a "snake" sprue. The numerical simulation results were compared with the experimental casting results, which were made on a centrifugal casting machine. The predicted shrinkage levels coincided well with the experimentally determined levels. Based on the above numerical analyses, an optimised running and gating system design for the crown model was proposed. The numerical filling and solidification results of the ball specimen showed that this simulation model could be helpful for the explanation of the experimentally indicated gas pores. It was concluded that shrinkage porosity in titanium dental casting was predictable, and it could be minimised by improving the running and gating system design. Entrapped gas pores can be explained from the simulation results of the mould filling and solidification. PMID- 10467948 TI - Journal of Pharmacy and Pharmacology and the electronic publishing revolution. PMID- 10467949 TI - Lipoplexes and tumours. A review. AB - The need for genotherapy to refocus its attention on to laboratory evaluation of better methods rather than proceeding to the clinic with semi-apt tools for genetic transfer has been highlighted in clinical study reports documented to date. Quintessential for tumour genotherapy is the ability to target abnormal cells, hence reducing exposure of normal cells to genetic material whilst maximizing gene dosage to tumour cells. This becomes increasingly important as genotherapy establishes itself in the clinic alongside the older modes of treatment. This review has discussed the applicability of lipoplexes for genotherapy of solid tumours. Lipoplexes have been used extensively for gene transfer into cells, such as cancerous cells, deficient for a certain gene product. While cationic liposomes have many advantages over other forms of delivery mechanisms, several problems hinder their use in-vivo. A closer examination of the physical limitations of current lipoplex preparations, the development and testing of novel formulations, combined with more attention to the cellular processes of cell membrane breaching and nuclear entry, may enhance gene delivery. Essential for tumour genotherapy is the ability to target these lipoplexes into tumour sites whilst reducing gene dosage to other normal tissues. Development of a better lipofection agent may indeed require a collaboration of the fields of physiology, cell biology, molecular biology, biochemistry, chemistry and membrane physics. PMID- 10467950 TI - Preparation, relative toxicity and therapeutic efficacy in mice and rats of liposomal HA-1-92, a new oxohexaene polyene macrolide antibiotic. AB - HA-1-92, a new polyene oxohexaene macrolide antibiotic isolated from Streptomyces CDRIL-312, was incorporated into liposomes containing phosphotidyl choline and cholesterol. The liposomal incorporated HA-1-92 considerably decreased toxicity when compared with free HA-1-92 in mice. Liposomal HA-1-92 showed improved pharmacokinetic profiles in rats. When administered to aspergillosis- and cryptococcosis-infected Balb/c mice, liposomal HA-1-92 showed increased antifungal activity, compared with free HA-1-92, with improved survival rate and decreased colony-forming units in lung, liver, spleen and kidney. These results suggest that liposomal HA-1-92 is more effective than free HA-1-92 in controlling experimental aspergillosis and cryptococcosis in Ba1b/c mice. PMID- 10467951 TI - Pharmacokinetics and pharmacological effect of recombinant human granulocyte colony-stimulating factor conjugated to poly(styrene-co-maleic acid) in rats. AB - A new derivative of recombinant human granulocyte colony-stimulating factor (rhG CSF) has been synthesized by conjugating rhG-CSF to poly(styrene-co-maleic acid) (poly(styrene-co-maleic acid)-rhG-CSF) to try to avoid glomerular filtration and thus potentiate the neutrophil-proliferating activity of rhG-CSF. Poly(styrene-co maleic acid)-rhG-CSF was highly bound to bovine serum albumin (BSA) and the molecular weight of the poly(styrene-co-maleic acid)-rhG-CSF-BSA complex was estimated to be about 90000 by gel filtration. Intravenous administration of poly(styrene-co-maleic acid)-rhG-CSF to normal rats resulted in a dose-dependent increase in neutrophil count. The neutrophil-proliferating activity of poly(styrene-co-maleic acid)-rhG-CSF was about 10 times greater than that of rhG CSF. After intravenous injection at a dose of 5 microg protein kg(-1) the total clearance of rhG-CSF fell from 71.0 to 32.1 mLh(-1) kg(-1) following poly(styrene co-maleic acid) modification. An isolated perfusion study in rat kidney showed that the filtered fraction of rhG-CSF was reduced by conjugation with poly(styrene-co-maleic acid). These results suggest that poly(styrene-co-maleic acid)-conjugation can potentiate the neutrophil-proliferating activity of rhG-CSF by reducing, at least in part, its renal clearance. PMID- 10467952 TI - Effect of fatty acids on the permeation of melatonin across rat and pig skin in vitro and on the transepidermal water loss in rats in-vivo. AB - Transdermal delivery of melatonin would be advantageous in the treatment of sleep disorders considering the short biological half-life of melatonin and its variable bioavailability via the oral route. This study looked at suitable penetration enhancers for the transdermal permeation of melatonin. The permeation of melatonin was enhanced by all saturated and unsaturated fatty acids across both rat and porcine skin. There was a parabolic relationship between the carbon chain length of saturated fatty acids and the enhancement of melatonin permeation across rat and porcine skin. For rat skin, the maximum flux was observed with undecanoic acid (45.33 microg cm(-2) h(-1)) which enhanced the flux of melatonin 8.6 times compared with the control, whereas lauric acid produced the maximum flux of melatonin (24.98 microg cm(-2) h(-1); 4-7 times) across porcine skin. An increase in the number of double bonds in cis-9-octadecanoic acid increased the flux of melatonin across rat skin. In contrast, with porcine skin, the flux of melatonin decreased as the number of double bonds increased, although the flux values were not statistically significant. Treatment of rats with undecanoic acid, oleic acid and linolenic acid for 3 h using Hill top chamber enhanced the transepidermal water loss significantly. The maximum transepidermal water loss was observed with undecanoic acid and linolenic acid among saturated and unsaturated fatty acids, respectively. Nonanoic acid and myristic acid did not cause a significant change in the transepidermal water loss. The enhancement effect of saturated fatty acids on the permeation of melatonin was dependent on the chain-length of the fatty acid in both rat and porcine skin. While an increase in the number of double bonds in the fatty acid increased the flux of melatonin in rat skin, no significant difference in the flux was observed with porcine skin. The permeation enhancement of melatonin by saturated and unsaturated fatty acids across rat skin was significantly higher than that of porcine skin. A positive correlation was observed between the permeation enhancement effect of the fatty acids across rat skin in-vitro and the transepidermal water loss in rats in-vivo, suggesting that there is a similarity in the mechanism by which fatty acids enhance the permeation of melatonin and in the enhancement of transepidermal water loss. We conclude that saturated fatty acids such as undecanoic acid or lauric acid which showed maximum permeation across rat and porcine skin, respectively, may be used as potential penetration enhancers in the development of a transdermal delivery system for melatonin. PMID- 10467953 TI - Cultured rabbit corneal epithelium elicits levofloxacin absorption and secretion. AB - Abstract Evidence for carrier-mediated transport of levofloxacin in the isolated rabbit cornea has been found. However, it is not known whether this mechanism is located in the epithelium or the endothelium. To resolve this question, we have measured the kinetics of levofloxacin uptake in primary cultures of rabbit corneal epithelial cells. The results indicate that levofloxacin accumulation was time dependent and a steady state was reached after 30 min. Maximal uptake occurred from a solution whose pH was 6.5. The uptake process was stereoselective and concentration dependent. In addition to the uptake, secretion of levofloxacin also occurred. These results indicate that the corneal epithelium is the site of levofloxacin transport mechanisms, mediating both absorption and secretion. PMID- 10467954 TI - Characterization of the carrier-mediated transport of levofloxacin, a fluoroquinolone antimicrobial agent, in rabbit cornea. AB - The cornea presents a formidable barrier to drug penetration. The fluoroquinolone levofloxacin, which is an effective antimicrobial agent, has the potential to be used in the topical treatment of ocular disease. Thus, we sought to characterize how levofloxacin penetrates the cornea. To perform this characterization, we measured the time dependent permeation of levofloxacin across the isolated rabbit cornea using a diffusion chamber, and compared it with antipyrine fluxes. Levofloxacin permeation into the receiver epithelial-side bathing solution (pH = 6.5) from the donor endothelial-side (pH = 7.4) reached 3.00 nmolcm(-2) cornea after 2h, whereas in the opposite direction permeation was 1.89 nmolcm(-2) cornea. Based on the temperature-dependent effects on permeation, the calculated energy of activation for permeation, Ea, was 31.3 kcal mol(-1), whereas Ea for antipyrine, a marker of diffusion, was 11.0 kcalmol(-1). The transport of levofloxacin from epithelium to endothelium was concentration-dependent and had both a linear and saturable component. Evaluation of the kinetic parameters, Jmax, apparent Km and k(d) showed that they were 38.78 pmol min(-1) cm(-2), 3.83 mM and 0.0135 microL min(-1) cm(-2), respectively. These results, coupled with the fact that levofloxacin permeation reached a maximum value at pH 6.5, suggest that levofloxacin transport across the cornea is carrier mediated. However, at present, it cannot be ascertained whether such a system is localized in either the corneal epithelial or the endothelial layer. PMID- 10467955 TI - The effect of pregnenolone 16alpha-carbonitrile on the pharmacokinetics and metabolism of dapsone in rats. AB - The purpose of this study was to evaluate the effect of pregnenolone 16 alpha carbonitrile (PCN) on the interconversion pharmacokinetics and metabolism of dapsone. To determine microsomal CYP3A activity and protein, eight rats (4 PCN, 4 corn oil) received a 1 mg kg(-1) intravenous bolus dose of dapsone, followed by blood and urine sampling. The formation clearance of dapsone hydroxylamine (CLf DDS-NOH) was calculated from the obtained samples. Interconversion pharmacokinetics estimates were obtained after 10 rats (5 PCN, 5 control) received 1 mg kg(-1) dapsone or 1.17 mg kg(-1) monoacetyldapsone, with a 24-h wash-out. Results from the interconversion analysis demonstrated that PCN significantly increased systemic clearance (CLs) of dapsone, but not its interconversion. The in-vivo/in-vitro correlation study demonstrated that PCN significantly increased CLs of dapsone (8.55 to 16.39mLmin(-1); P<0.01) and CLf DDS-NOH (0.13 to 0.18mLmin(-1); P<0.01). PCN treatment produced a 69% increase in CYP3A protein, and increased 6beta- and 2beta-hydroxytestosterone formation rates. Significant correlations were found between CLf DDS-NOH and either 6beta- (r2 = 0.925), 2beta-hydroxytestosterone (r2 = 0.92), or CYP3A1/2 protein (r2= 0.60). We conclude that PCN treatment produces significant increases in CLs (dapsone) and CLf (DDS-NOH) in rats. These changes were not due to changes in the reversible metabolism of dapsone. These results suggest that the formation clearance of dapsone hydroxylamine reflects alterations in CYP3A activity, despite the fact that it accounted for a small part of the systemic clearance of dapsone. PMID- 10467956 TI - Comparison of cyclosporin A and tacrolimus concentrations in whole blood between jejunal and ileal transplanted rats. AB - Most immunosuppresive drugs are absorbed from the intestine after oral administration, although there is some difference of bioavailability between ileum and jejunum. Using an orthotopic segmental small bowel transplantation (SBT) model in rats, we studied the pharmacokinetic profiles of cyclosporin A and tacrolimus concentrations after oral intake, comparing jejunal and ileal transplanted rats. Two types of segmental SBT (jejunal and ileal SBT) in a syngeneic combination were performed. After oral administration of cyclosporin A (10mgkg(-1) or tacrolimus (5 mg kg(-1)), pharmacokinetic data were obtained from the long-surviving rats transplanted with segmental SBT. To determine the effect of additional bile on cyclosporin absorption, an emulsion of cyclosporin A with fresh bile juice was re-challenged on segmental SBT rats before killing. A histological study was also performed by use of the intestinal grafts from the killed SBT rats. A higher concentration of cyclosporin A was observed in the ileum-grafted rats than in the rats which received the jejunal grafts. Oral bioavailability of cyclosporin A in ileal SBT rats tended to be increased by addition of fresh bile juice, but that in jejunal SBT rats did not change. On the other hand, there was no significant difference of tacrolimus concentration between jejunum- and ileum-transplanted rats. Histological studies showed that the superficial mucosal layer of both grafts, but especially the ileal graft, was markedly elongated compared with that of normal intestine. The present study showed that cyclosporin A was more actively absorbed from ileum than from jejunum in SBT, but tacrolimus was absorbed equally from both sites. These data suggest that cyclosporin A concentration is satisfactorily controlled in the segmental ileal graft, while there is no difference of tacrolimus absorption between ileal and jejunal graft. PMID- 10467957 TI - Effect of caffeine on ibuprofen-induced gastric mucosal damage in rats. AB - During investigations on the effect of caffeine on ibuprofen-induced gastric mucosal lesions in rats, we have found that caffeine (p.o.) inhibits the development of ibuprofen-induced gastric lesions in a dose-dependent manner (ED50 18.4 mg kg(-1)). To investigate this protective effect of caffeine, we have studied the effect of caffeine on HCl-ethanol-induced gastric mucosal lesions with or without indomethacin pretreatment. Caffeine inhibited the development of HCl-ethanol-induced gastric lesions with and without indomethacin pretreatment. These results indicate that caffeine did not act as a mild irritant but, on the contrary, had protective effects. We measured the gastric mucosal prostaglandin E2 (PGE2) concentrations and gastric mucosal blood flow, as representative protective factors for gastric mucosa. Caffeine did not affect the gastric mucosal PGE2 concentrations 4h after administration of ibuprofen. However, topical administration of caffeine resulted in an increase in gastric mucosal blood flow, as measured by laser Doppler flowmetry. We investigated the gastric acid secretion and gastric mucosal myeloperoxidase activity as representative aggressive factors for gastric mucosa. When caffeine was administered intraduodenally in pylorus-ligated rats, gastric acid secretion decreased in a dose-dependent manner, with an ED50 of 44.9 mg kg(-1). Caffeine decreased ibuprofen-induced gastric myeloperoxidase activity in a dose-dependent manner, with an ED50 of 9.1 mg kg(-1). These findings indicate that caffeine, at least in rats, may inhibit the development of acute gastric mucosal injury. The mechanisms underlying the protective actions of caffeine are unclear, but may be related in part to an increase in gastric mucosal blood flow and suppression of neutrophil activation. PMID- 10467958 TI - The therapeutic effect of proton pump inhibitors on Helicobacter pylori-positive gastric ulcers. AB - The aim of the present study was to elucidate the risk factors that could delay gastric ulcer healing when either a proton pump inhibitor or an H2-receptor antagonist is used for gastric ulcer treatment. Endoscopically-diagnosed gastric ulcer patients (216 men and 96 women, mean age: 57+/-13 years) were investigated. All patients were consecutively recruited and randomly assigned to receive H2 receptor antagonist (n = 196) or proton pump inhibitor (n = 116) treatment for eight weeks. Chi-squared tests and multivariate analysis to determine factors influencing ulcer healing were used to analyse the patients profiles, endoscopic findings, and Helicobacter pylori-infection status. In the H2-receptor antagonist group, the most important risk factor was a large ulcer size (> 2 cm diam.), followed by a linear shape of the ulcer, undermining tendency of ulcer, previous history of gastric ulcer, and H. pylori infection. In the proton pump inhibitor group, linear shape of the ulcer was the only significant risk factor for slow ulcer healing; other factors, including H. pylori infection, were insignificant. These results indicate that ulcer morphology may be the most important information for predicting ulcer healing, and that H. pylori infection does not delay gastric ulcer healing when proton pump inhibitor treatment is used. PMID- 10467959 TI - Possible involvement of Ca2+ entry and its pharmacological characteristics responsible for endothelium-dependent, NO-mediated relaxation induced by thapsigargin in guinea-pig aorta. AB - Thapsigargin, a specific inhibitor of Ca(2+)-pump Ca(2+)-ATPase in the sarcoplasmic/endoplasmic reticulum (SR/ER), produces an endothelium-dependent vascular relaxation. In the present study, pharmacological features of thapsigargin-induced endothelium-dependent relaxation were functionally characterized in the isolated guinea-pig aorta especially focusing on the Ca2+ mobilization mechanisms in endothelial cells. Thapsigargin-induced endothelium dependent vascular relaxation was markedly suppressed by N(G)-nitro-L-arginine (L NNA) and calmidazolium, suggesting that the vascular relaxation to thapsigargin is largely attributable to endothelium-derived nitric oxide (NO) produced as a result of the activation of Ca2+, calmodulin-dependent NO synthase (NOS). Removal of Ca2+ from the external solution abolished the endothelium-dependent relaxation of guinea-pig aorta in response to thapsigargin. Thapsigargin-induced endothelium dependent relaxation was inhibited more strongly compared with the endothelium independent relaxation to an NO donor, SIN-1 (3-(4-morpholinyl)-sydnonimine), when the artery preparation was preconstricted with a high concentration (80 mM) of KCl instead of agonistic stimulation. Endothelium-dependent relaxation induced by thapsigargin was not affected by diltiazem, a blocker of L-type voltage-gated Ca2+ channels. SK&F96365 (1-[beta-[3-(4-methoxyphenyl)propoxy]-4 methoxyphenethyl]-1 H-imidazole) and Ni2+, both of which block capacitative Ca(2+) entry, did not show any appreciable inhibitory effects on the endothelium dependent relaxation to thapsigargin. These findings suggest that in guinea-pig aorta, endothelium-dependent NO-mediated relaxation induced by thapsigargin is preceded by the increase in the cytosolic free Ca2+ concentrations ([Ca2+]cyt) following the depletion of stored Ca2+ in thapsigargin-sensitive store sites in endothelial cells. Although the increase in [Ca2+]cyt responsible for the activation of endothelium NOS leading to thapsigargin-induced vascular relaxation may be ascribed to the capacitative Ca2+ entry from extracellular space, the Ca2+ entry mechanism stimulated with thapsigargin is deficient in sensitivity to SK&F96365 and Ni2+ in the endothelium of guinea-pig aorta. PMID- 10467960 TI - A mild transient decrease of peripheral red blood cell counts induced by a suprapharmacological dose of pegylated human megakaryocyte growth and development factor in rats. AB - Previous studies have shown that pegylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) at suprapharmacological dose induces a mild transient decrease of red blood cell counts according to thrombopoiesis in normal mice. To unravel the mechanism underlying this mild transient decrease of red blood cells, we have studied the effect of PEG-rHuMGDF on the circulating plasma and blood volume, and the serum biochemical parameters of anaemia and splenectomy. Also, we have performed histological studies of the bone marrow and the spleen of PEG-rHuMGDF-treated rats. PEG-rHuMGDF (300 microg kg(-1)]) or vehicle was subcutaneously administered to rats once a day for up to five days. From day 6 after the start of PEG-rHuMGDF administration, the platelet counts and plateletcrit levels were significantly increased, reaching peak values on day 10, and recovering to normal by day 20. The red blood cell counts and the haematocrit levels were significantly decreased on day 6 to 13. The decreases in red blood cell levels and haematocrit produced by PEG-rHuMGDF treatment were mild and had recovered by day 15. The plasma and blood volumes were significantly increased on day 10 in PEG-rHuMGDF-treated rats. No alteration of the serum biochemical parameters for anaemia, iron or total bilirubin, were observed on day 10. The histological examination on day 10 revealed a marked increase in megakaryocytes and a slight decrease in erythropoiesis in the bone marrow of rats that received PEG-rHuMGDF (300 microg kg(-1)). There was also a slight increase in splenic megakaryocytes and erythropoiesis. The decrease of red blood cells by PEG-rHuMGDF was not affected by splenectomy. These results suggest that the mild transient decrease of red blood cells induced by PEG-rHuMGDF treatment for up to five days is based mainly on the increases in the plasma and blood volume. These events are secondary changes due to the regulation of the excess production of megakaryocytes in the marrow and the peripheral platelets. PMID- 10467962 TI - Muscle relaxant activity of methocarbamol enantiomers in mice. AB - Documented studies support the emerging idea that drug enantiomers could have different pharmacological activity. Our bibliographical data have shown that so far no report has been published on the pharmacological activity of individual enantiomers of methocarbamol. This study was conducted to characterize the muscle relaxant activity of methocarbamol enantiomers. The rotarod test was used to compare the muscle relaxant activity of racemic methocarbamol and pure enantiomers after intraperitoneal administration of the enantiomers to mice. The results show that (+)-R-methocarbamol has higher muscle relaxant activity compared with racemic methocarbamol or (-)-S-methocarbamol. PMID- 10467961 TI - Choline uptake by mouse brain capillary endothelial cells in culture. AB - Choline, a precursor of the neurotransmitter acetylcholine, is synthesized in only small amounts in the brain, so the choline concentration in the brain may vary depending on the plasma concentration and the transport rate across the blood-brain barrier. To elucidate the transport mechanism of choline, we carried out uptake experiments with mouse brain capillary endothelial cells in culture (MBEC4). [3H]Choline uptake was linear for up to 5 min. An examination of the concentration dependence of [3H]choline uptake revealed the operation of both saturable (Jmax = 423+/-27 pmol min(-1) (mg protein)(-1) and Kt = 20.0+/-3.1 microM) and non-saturable (kd = 1.23+/-0.045 microL min(-1)(mgprotein)-1) processes. The saturable process was independent of Na+ and pH, but was dependent on membrane potential as a driving force. Various basic drugs and endogenous substances, including substrates and inhibitors of the organic cation transporter, significantly inhibited the [3H]choline uptake. These data suggest that choline was taken up into the endothelial cells via two routes and that a membrane potential-dependent carrier-mediated transport system may participate in choline transport across the blood-brain barrier. PMID- 10467963 TI - Effects of a novel pyridylsulphonyl thiazole derivative, FR115092, on autoimmune and mitomycin C-induced thrombocytopenia in mice. AB - Dapsone (4,4'-diaminodiphenyl sulphone), an antileprotic and antimalarial drug, has been reported to be of therapeutic benefit in idiopathic thrombocytopenic purpura in the clinic. However, adverse reactions such as haemolytic anaemia have often been observed. In this study, we found that dapsone increased the number of platelets and decreased the number of red blood cells in male (NZWxBXSB)F1 (W/BF1) mice, an animal model of idiopathic thrombocytopenic purpura. In studies to prepare derivatives of dapsone with weaker side effects than the parent compound, FR115092 (2-[5-(2-pyridylsulphonyl)thiazolyl]amine) was discovered. The effect of FR115092 on the number of blood cells was studied and compared with dapsone in mice. FR 115092 increased the number of platelets without reducing the number of red blood cells in W/BF1 mice. This drug significantly suppressed the increase in circulating autoantibodies against platelets and increased the number of megakaryocytes. Furthermore, FR115092 inhibited the reduction of the number of platelets in mitomycin C-induced thrombocytopenic mice, as a consequence of its enhancement of growth and maturation of megakaryocytes. These findings suggest that FR115092 may be effective against various thrombocytopenias, without inducing haemolytic anaemia. PMID- 10467964 TI - Oral administration of sepimostat mesilate prevents acute alcohol pancreatic injury in rats. AB - The preventive effect of a novel synthetic serine protease inhibitor, sepimostat mesilate (sepimostat), on acute alcohol pancreatic injury, induced by exocrine hyperstimulation and ethanol administration, was assessed and compared with that of a similar protease inhibitor, camostat mesilate (camostat). Conscious rats were infused with 1 microg mL(-1) h(-1) caerulein intravenously for 6 h and with 0.1 g mL(-1) h(-1) ethanol for 9 h, with the latter infusion beginning 3 h after the start of the caerulein infusion. Sepimostat or camostat was administered orally 1 h before the caerulein infusion. Rats infused with caerulein plus ethanol showed increased plasma amylase and lipase activities, and aggravated pancreatic interstitial oedema when compared with rats given caerulein alone. Sepimostat at 10 and 30 mg kg(-1) prevented the increase in plasma amylase and lipase activities caused by caerulein plus ethanol infusion. Sepimostat at 30 mg kg(-1) suppressed the histological change. Camostat did not show any preventive effects at the equivalent dose. When conscious rats were infused with 1 microg mL(-1) h(-1) caerulein alone intravenously for 6 h, plasma amylase and lipase activities were increased compared with rats given saline. Neither drug prevented the increase in these activities at 30mg kg(-1). Our results suggest that sepimostat has superior preventive effects on alcohol-induced acute pancreatic injury compared with camostat. Sepimostat may thus be a useful drug in the therapy of alcohol-induced pancreatitis. PMID- 10467965 TI - A new HPLC technique for the separation of methocarbamol enantiomers. AB - We have developed a stereoselective high-performance liquid chromatography technique for analytical separation of methocarbamol enantiomers. Precolumn derivatization was performed at room temperature using (-)-menthylchloroformate as a chiral reagent in the presence of pyridine as catalyst. The resulting diastereomers were separated on two Resolve C18 columns connected in series. The mobile phase was phosphate buffer (pH 7.5)-acetonitrile (50: 50, v/v) at a flow rate of 1 mL min(-1). UV detection was set at 274 nm. The optimum amount of reagent and the maximum peak intensity of the diastereomers were determined. The resolution of the diastereomers was satisfactory (alpha = 1.04) under the conditions used. PMID- 10467966 TI - Pharmacotherapy response and diagnostic validity in atypical depression. AB - BACKGROUND: The validity of diagnostic criteria and the efficacy of tricyclic antidepressant pharmacotherapy for atypical depression were studied in the NIMH Treatment of Depression Collaborative Research Program. METHODS: Outpatients with major depressive disorder (N = 239) entered a 16-week clinical trial and were randomly assigned to interpersonal psychotherapy, cognitive behavior therapy, and imipramine or placebo with clinical management. Features of atypical depression were rated on the SADS and ISI and clinical outcome was measured on the HRSD and GAS. RESULTS: Atypical features of mood reactivity and at least one reversed vegetative symptom of hypersomnia, hyperphagia or weight gain (25.2% patients) were predictive of pharmacotherapy non-responsiveness with imipramine compared to placebo. The additional features of diurnal mood variation, 'leaden paralysis', and 'rejection sensitivity' did not further distinguish animipramine non responsive subgroup. Imipramine did show significant effectiveness compared to placebo among non-atypical patients on measures of depressive symptom change. LIMITATIONS: The predictive influence of atypical features was not accounted for on the basis of depression severity. CONCLUSIONS: This study provides evidence for the predictive validity of atypical features of major depressive disorder, including mood reactivity and at least one reversed vegetative symptom of either hypersomnia, hyperphagia, or weight gain, supporting the inclusion of atypical depressive features, with these criteria, in the DSM-IV. PMID- 10467967 TI - A comparison of the age of onset of bipolar I and bipolar II outpatients. AB - BACKGROUND: It is not clear if bipolar I and bipolar II disorders are variants of the same disorder or distinct disorders. Aim of the study was to compare age at onset and indicators of severity (recurrences, chronicity, psychosis) between bipolar I and bipolar II disorders to clarify this point. METHODS: Consecutive 45 bipolar I and 141 bipolar II outpatients were interviewed with the Structured Clinical Interview for DSM-IV. Age at onset distributions were compared with the Kolmogorov-Smirnov test. RESULTS: Age at onset distributions were not significantly different between bipolar I and bipolar II patients. Severity was significantly higher in bipolar I patients. LIMITATIONS: Age at onset may be subject to recall bias. Structured interview by a senior psychiatrist, standard criteria for onset, family members or close friends supplementing information, same data collection and ascertainment procedures may have reduced bias. CONCLUSIONS: Lack of significant difference in age at onset does not support the separation of the two disorders. Bipolar II disorder may be a less severe variant of bipolar disorder. PMID- 10467968 TI - A comparison of clinical response to electroconvulsive therapy in puerperal and non-puerperal psychoses. AB - BACKGROUND: There is a widespread belief that puerperal psychosis is particularly responsive to electroconvulsive therapy (ECT), but research evidence for this is lacking. METHOD: We have conducted a retrospective case-note study of clinical improvement following ECT in puerperal and non-puerperal psychosis. The main outcome measure was improvement in mental state at the end of a course of ECT, rated by a blind rater on a simple four-point scale. Additional indicators of responsiveness to ECT were improvement in mental state 4 weeks after stopping ECT, duration of in-patient stay following ECT and number of ECT received. RESULTS: Women with puerperal psychosis showed greater clinical improvement than women with non-puerperal psychosis. The results were not explained by the greater preponderance of depressive illness in the puerperal group, as the same results were also found when the analysis was confined to women with a clinical diagnosis of depressive illness. CONCLUSION: These findings are the first evidence of a particular sensitivity of ECT in puerperal psychosis. However, they are preliminary and a number of explanations are possible, including good responsiveness to treatment in general. A prospective study using standardised clinical ratings and definitions of key variables is now required. CLINICAL IMPLICATIONS: Clinicians treating severe postpartum illness should continue to regard ECT as a treatment option. LIMITATIONS OF THE STUDY: The study is limited by its reliance on retrospective examination of information recorded in case notes. Ratings of clinical improvement were not standardised. PMID- 10467969 TI - Endogenous digoxin-like immunoreactive factor (DLIF) serum concentrations are decreased in manic bipolar patients compared to normal controls. AB - BACKGROUND: A decrease in sodium pump activity in erythrocytes has been associated with manic episodes of bipolar illness relative to euthymic moods. Since red blood cells are long-lived and lack a nucleus, it is likely that a plasma factor is responsible for the observed decrease in sodium pump activity. METHODS: Utilizing a radioimmunoassay, we examined the serum concentrations of the digoxin-like immunoreactive factor (DLIF) in ill and well bipolar patients and compared the values to those of normal controls. RESULTS: DLIF was significantly decreased in manic individuals as compared to normal controls (143.6+/-S.E.M. 20.94 vs. 296.6+/-12.76 pg digoxin equivalents/ml, respectively, F = 4.77, P<0.05), but not compared to euthymic bipolar subjects 213.8+/-86.92, P = 0.77). There were no significant differences in DLIF concentrations between manic and euthymic bipolar individuals (P = 0.8). Since relapse in bipolar patients appears to display a seasonal pattern, we also measured the plasma concentration of this factor over a 12-months period. Normal controls exhibited a seasonal pattern of change in serum DLIF concentrations with a nadir in the winter months. Plasma concentrations of DLIF in bipolar patients did not show a seasonal pattern and maintained low levels throughout the year. LIMITATIONS: Due to the nonspecificity of our antibody, we could measure only total DLIF. Furthermore, it is unclear what the role of circulating DLIF, if any, may be on brain function. CONCLUSION: DLIF may be involved in the pathophysiology of mania. PMID- 10467970 TI - Prediction of response to fluoxetine and placebo in children and adolescents with major depression: a hypothesis generating study. AB - BACKGROUND: The results of multivariate analyses to identify potential predictors of response to fluoxetine or placebo separately in 96 child and adolescent outpatients with major depressive disorder from a recent controlled trial are presented. METHODS: A variety of clinical, demographic and laboratory factors were examined as possible predictors of response to fluoxetine or placebo using logistic regression models. RESULTS: No single variable or combination of variables strongly predicted response to fluoxetine. For the placebo group, a younger age, a shorter duration of depressive episode, and a lower socioeconomic status predicted response with an overall predictive power of 81%. CONCLUSIONS: This study is limited by the small sample size and should be considered hypothesis generating rather than confirming. PMID- 10467971 TI - Avoidant personality in social phobia and panic-agoraphobic disorder: a comparison. AB - BACKGROUND: Avoidant personality disorder (APD) is generally believed to be related to social phobia (SP), especially to generalized subtype. However, it has also been reported to be prevalent in panic disorder-agoraphobia (PDA). In the present investigation, we wished to explore whether APD in each of these disorders has discriminatory features. METHOD: We studied 71 SP and 119 PDA patients with state-of-the-art clinical instruments based on DSM-III-R. RESULTS: The pattern of social avoidance in SP was more pervasive: it was characterized by a higher level of interpersonal sensitivity and greater severity, associated with psychopathology as well as a higher rate of Axis I comorbidity. By contrast, avoidance of non-routine situations characterized APD occurring in the setting of PDA. LIMITATIONS: Differences in inclusion criteria and comorbidity rates, as well as overlap between different operational disorders, may have influenced our findings. CONCLUSION: ADP is operationally broad, and 'avoidant' as a specifier of a personality type is insufficiently precise. ADP captures avoidant traits - which appear secondary to a core dimension such as interpersonal sensitivity--but is basically a heterogeneous condition influenced by the nature of comorbid Axis I disorders. PMID- 10467973 TI - Outcome in refractory depression. AB - BACKGROUND: Failure to respond to first-line antidepressant treatment can occur in up to 40% of patients with depressive illness. A proven strategy for managing this refractory depression is lithium augmentation. The long-term outcome and optimal management of patients treated with lithium augmentation remains unclear. We describe a 4-8 year naturalistic follow-up of patients treated with lithium augmentation in two controlled studies of its efficacy in refractory depression. METHOD: Cases were followed up with personal interview where possible, and by telephone and general practitioner contact otherwise. Lifetime clinical status was ascertained using the Schedule for Affective Disorders and Schizophrenia Lifetime (SADS-L). RESULTS: We obtained outcome data on 53 of the original eligible 76 patients. There was a good outcome in 38 (72%) patients. Good outcome was associated with a less endogenous nature of depression and an absence of previous hospitalisations. CONCLUSIONS: There do not seem to be any specific prognostic indicators of long-term outcome to lithium augmentation beyond those recognised to be relevant in the outcome of depression generally. LIMITATIONS: The conclusions are limited by incomplete follow-up of the total original sample and lack of objective illness and medication data for the intervening period. PMID- 10467972 TI - Six months of desipramine for dysthymia: can dysthymic patients achieve normal social functioning? AB - BACKGROUND: There is evidence that antidepressant medication improves social dysfunction during acute treatment in dysthymic patients but it is unknown if the gain in social functioning persists or progresses with longer-term antidepressant treatment. We examine the effect of 6 months of desipramine treatment on social functioning in dysthymic patients. METHODS: Forty-six subjects with DSM-III-R dysthymia (70% with superimposed major depression) who had responded to 10 weeks of open-label desipramine (DMI) treatment received 16 additional weeks of continuation DMI. Social functioning was measured at weeks 0, 10 and 26 with the Social Adjustment Scale-Self Report. RESULTS: Euthymia was maintained and a marginally significant trend for further improvement in overall social functioning appeared during continuation treatment. Only 24% of subjects achieved normative level of social adjustment after 6 months of DMI treatment. LIMITATIONS: The main limitation was the lack of a placebo control group. CONCLUSION: Acute improvement in social functioning persists during continuation treatment. However, most dysthymic patients did not achieve a community level of social adjustment. Significant social dysfunction persists in dysthymic patients with low levels of depressive symptomatology after 6 months of intense DMI treatment. PMID- 10467974 TI - Hypothermic, ACTH, and cortisol responses to ipsapirone in patients with mania and healthy controls. AB - BACKGROUND: The selective 5-HT1A receptor agonist ipsapirone causes dose dependent decrease in body temperature and increase in adrenocorticotropic hormone (ACTH) and cortisol release in humans. These responses are attenuated by 5-HT1A receptor antagonists, suggesting that hypothermia, ACTH and cortisol release induced by ipsapirone are indeed mediated by 5-HT1A receptors and that these responses provide a valid index of 5-HT1A receptor function in humans. METHODS: To examine the 5-HT1A receptor sensitivity in patients with mania, we studied six manic patients and six age and sex matched healthy controls. After obtaining a blood sample for baseline hormone levels and measuring body temperature, a single dose of 0.3 mg/kg of ipsapirone was given orally to all the subjects and further bloods and temperature reading were obtained every 30 minutes for 3 hours. RESULTS: We found that ACTH and cortisol responses to ipsapirone were significantly increased in mania when compared to healthy controls, but there was no significant difference in hypothermic response to ipsapirone between the two groups. LIMITATIONS: A lack of placebo control, heterogeneity of patients, and a small sample size are the limitations. CONCLUSIONS: Our findings suggest that manic patients may have enhanced postsynaptic 5-HT1A receptor sensitivity, but presynaptic 5-HT1A receptors are unaltered in this condition. Further placebo-controlled studies with a larger number of manic patients are needed to verify this. PMID- 10467975 TI - Prevalence of dysthymic disorder in primary care. AB - BACKGROUND: Dysthymic disorder is characterised as a chronic state of depressed mood which is not otherwise attributable to physical, psychological or social events. While it can occur alone, there is increasing evidence that the majority of individuals who meet criteria for dysthymic disorder also experience more severe episodic mood disorders throughout their lifetime, and there is also an aggregation of mood disorders within their family members. Patients with dysthymic disorder are most often seen in primary care. Some researchers suggest that the majority of these individuals are never diagnosed or are not diagnosed until a more severe episodic mood disorder develops. The objective of this study was to determine the 12-month prevalence of Axis I psychiatric disorders, and in particular dysthymic disorder, in a primary care Health Service Organization in Ontario, Canada. METHODS: Eligible and consenting adults registered with a primary care Health Service Organization were screened using the modified form of the University of Michigan Composite International Diagnostic Interview. RESULTS: Of the 6280 eligible subjects, 4327 (69%) consented to screening. Two hundred and twenty-two (5.1%) subjects screened positive for dysthymic disorder. In addition, 90% of those who screened positive for dysthymic disorder also screened positive for other Axis I disorders including major depressive disorder, panic, simple phobia, and generalized anxiety disorder. CONCLUSIONS: There is much potential for the primary care physician to play a pivotal role in the recognition and treatment of dysthymic disorder and associated Axis I disorders. A focus on the family as a unit for care may be especially important given the reported aggregation of dysthymic disorder within families. PMID- 10467976 TI - Guanfacine and secondary mania in children. AB - INTRODUCTION: Guanfacine hydrochloride is an alpha-2 adrenergic agonist, which has gained recent attention in the field of child and adolescent psychiatry. This medication has been described as effective in the management of attention-deficit hyperactivity and tic disorders, with minimal side effects. METHODS: Presented here are five cases of behavioral activation in children treated with guanfacine. RESULTS: In each instance the clinical presentation resembled an acute hypomanic or manic episode. The dose of guanfacine was 0.5 mg/day. Later investigation revealed that all of the youngsters had clear risk factors (clinical and/or familial) for bipolar disorder. CONCLUSIONS: It appears as though guanfacine may be capable of precipitating secondary mania in vulnerable children. PMID- 10467977 TI - Gabapentin as a promising treatment for antipsychotic-induced movement disorders in schizoaffective and bipolar patients. AB - Improvement of antipsychotic-induced blepharospasm and involuntary oral-mandibulo movements was observed with the use of the anticonvulsant drug gabapentin among 14 of 16 affectively ill patients who had been exposed to maintenance neuroleptics of the conventional type. In many cases, the movement disorders of these patients had not responded to more standard measures such as clozapine. This finding permits a potential strategy for patients with treatment-emergent tardive dyskinesia, a well-known complication of extended conventional neuroleptic use. Gabapentin, whose mood stabilizing properties have been reported in several clinical reports, represents a more natural treatment in the setting of bipolar spectrum disorders. PMID- 10467978 TI - The bipolar spectrum: a clinical reality in search of diagnostic criteria and an assessment methodology. AB - Failure to recognize subthreshold expressions of mania contributes to the frequent under-diagnosis of bipolar disorder. There are several reasons for the lower rate of recognition of subthreshold manic symptoms, when compared to the analogous pure depressive ones. These include the lack of subjective suffering, enhanced productivity, ego-syntonicity, and diurnal and seasonal rhythmicity associated with many of the manic and hypomanic symptoms, and the psychiatrists' tendency to subsume persistent or even alternating symptoms among personality disorders. Furthermore, the central diagnostic importance placed on alterations in mood distracts clinicians from paying attention to other more subtle but clinically meaningful symptoms, such as changes in energy, neurovegetative symptoms and distorted cognitions. Although officially accepted in both ICD-10 and DSM-IV, we believe bipolar II disorder is underdiagnosed because of inattention to symptoms of hypomania. Moreover, by requiring the presence of both full-blown hypomanic and major depressive episodes, current nosology fails to include symptoms or signs which are mild and do not meet threshold criteria. There is already agreement in the field that such symptoms are important for depression. We now propose that attention should also be devoted to mild symptomatic manifestations of a manic diathesis, even if such manifestations may sometimes enhance quality of life. The term 'spectrum' is used to refer to the broad range of such manifestations of a disorder from core symptoms to temperamental traits. Spectrum manifestations may be present during, between, or even in the absence of, an episode of full-blown disorder. We have developed a structured clinical interview to assess the mood spectrum (SCI-MOODS) to evaluate the whole range of depressive and manic symptoms. This instrument is currently undergoing psychometric testing procedures. Similar to the SCID interview, the SCI-MOODS interview provides a separate rating for each of the major DSM-IV symptoms, but the latter also identifies and rates subthreshold and atypical manifestations. This paper presents the concept of a subthreshold bipolar disorder and discusses the potential epidemiological, diagnostic and therapeutic relevance of such a spectrum conditions. We also describe the SCI-MOODS interview used reliably to identify the occurrence of a bipolar spectrum condition. Obviously a great deal of systematic research needs to be conducted to ascertain the reliability and validity of subthreshold bipolarity as summarized in this paper and embodied in our instrument. PMID- 10467979 TI - Dominant regulation: a common mechanism of monoclonal antibody induced tolerance? AB - Transplantation tolerance can be induced by a range of agents that block T cell/antigen-presenting cell (APC) interactions known to be important for initiation of the adaptive immune response. Tolerance so induced has been shown to have a regulatory phenotype dependent on CD4+ cells. This was first observed with nonlytic anti-CD4 antibodies, and was recently demonstrated following other therapeutic approaches. Dominant tolerance also plays a role in natural regulation of the immune response, functioning to prevent autoaggressive cells mediating self-destruction. The mechanism by which dominant tolerance is established and maintained remains unclear, and the reported characteristics of regulatory cells in different experimental models vary widely. Here we review the evidence for potential mechanisms involved and propose that there is a common pathway by which dominant tolerance is mediated. PMID- 10467981 TI - Small GTPases in lymphocyte biology: Rho proteins take center stage. AB - Almost a decade ago, the small GTPase Ras was shown to be activated in response to antigen receptor triggering in T cells. Since then, Ras has been further characterized as a central molecule for the regulation of signal transduction pathways in lymphocytes. However, over the last couple of years, its exclusive role in lymphocyte biology has been challenged by the emergence of its relatives of the Rho family. Today it is well established that Rho GTPases act as unique molecular switches at several critical checkpoints in lymphocyte development and function. Additionally, a new and critical concept in GTPase signaling has taken shape over the last couple of years in that small GTPases are able to regulate quite diverse cellular processes in the immune response by linking to multiple biochemical effector pathways. PMID- 10467980 TI - Role of cytokines in pulmonary antimicrobial host defense. AB - Host defense of the lung is characterized by a fine balance between the generation of a vigorous inflammatory response to clear pathogens and maintenance of the integrity of the alveolar gas-exchange surface. The magnitude of the inflammatory response is therefore tightly regulated by pro- and anti inflammatory cytokine mediators. This article summarizes current information on the roles of specific cytokines in pneumonia, with particular emphasis on ongoing investigations into the role of innate immunity in bacterial and fungal pneumonia. PMID- 10467982 TI - Developmental regulation of immunoglobulin mRNA processing and the IgA response: establishing a paradigm. AB - IgA, which is protective at mucosal sites, is derived from memory B cells that develop in the organized lymphoid tissue of the gastrointestinal tract and subsequently mature to plasma cells in the lamina propria. Similarly to B cells expressing other isotypes, the maturation of IgA-expressing B cells is associated with both an increase in the steady-state level of immunoglobulin mRNA and the ratio of secreted to membrane forms of mRNA, which differ in 3' terminus. In contrast to B cells expressing other isotypes, at all stages in the development of an IgA response, the secreted form of alpha mRNA predominates. In this article, studies on the general features of IgA B cell development, mechanisms regulating 3' terminus usage of Ig mRNAs, and isotype-specific regulation of 3' terminus usage particularly in regard to alpha mRNA are discussed. PMID- 10467983 TI - The development of CD4+ T effector cells during the type 2 immune response. AB - Multiple pathways may be involved in the development of interleukin 4 (IL-4) producing T helper (Th) cells and the associated type 2 immune response. Increasing evidence suggests that the strength of signals delivered to the T cell may favor the development of the type 2 response. In contrast, antigen-presenting cell- (APC) derived stimuli produced following pattern recognition receptor binding during the innate response promotes the development of interferon-gamma (IFN-gamma) producing cells and the associated type 1 immune response. In many cases, the balance between increased signaling strength and the innate response may determine whether the type 2 response develops. T cell receptor (TCR), CD4, and costimulatory molecule interactions may all contribute to signal strength, but the type 2 immune response may be particularly dependent on the availability of coreceptor and costimulatory molecule interactions. B7 ligand interactions are required for the development of the type 2 immune response and interaction of CD28 with either B7-1 or B7-2 can provide sufficient signals for its initiation. In B7-2-deficient mice, the initial type 2 immune response is intact, but the response is not sustained, suggesting that B7-2 is important at later stages of the type 2 immune response. The roles of CD28 and CTLA-4 during the type 2 response remain unclear. The type 2 response to infectious pathogens is pronounced in CD28-/- mice, suggesting that other costimulatory molecule interactions can substitute for CD28 for the development of IL-4 producing T cells and the associated type 2 immune response. PMID- 10467985 TI - The threat of therapeutic Nihilism. PMID- 10467986 TI - Food-borne and water-borne infections and the gastroenterologist. PMID- 10467984 TI - Murine mercury-induced autoimmunity: a model of chemically related autoimmunity in humans. AB - Human exposure to certain compounds or therapeutic drugs can result in the development of an autoimmune syndrome. Mercury (Hg) induced autoimmunity is one of the few animal models in which administration of a chemical induces a specific loss of tolerance to self-antigens. After receiving subtoxic doses of Hg or other heavy metals, susceptible mouse strains rapidly develop highly specific antibodies to nucleolar antigens. In addition, these animals display a general activation of the immune system, especially pronounced for the Th2 subset and a transient glomerulonephritis with immunoglobulin deposits. Like many human autoimmune diseases, this syndrome is associated with the expression of susceptible major histocompatibility complex (MHC) class II genes. In this article, we review the essential features of this model, and we discuss the putative mechanisms by which Hg creates such a severe immune dysfunction. PMID- 10467987 TI - Novel strategies for preventing diarrheal diseases. PMID- 10467988 TI - Role of transjugular intrahepatic portosystemic shunt in the treatment of portal hypertension in pediatric patients. PMID- 10467989 TI - The leptin era: new insight into the mechanisms of body weight homeostasis. PMID- 10467990 TI - Shwachman's syndrome: pathomorphosis and long-term outcome. AB - BACKGROUND: Shwachman's syndrome is the second most common cause of inherited/congenital pancreatic insufficiency after cystic fibrosis. The main associated features are usually cyclic neutropenia, metaphyseal dysostosis, and growth retardation. Other organs or functions may be involved in this syndrome, showing a wide range of abnormalities and symptoms. There are reports of Shwachman's syndrome in childhood, but little is known about the long-term clinical course of these patients. This article reports on the pathomorphosis and long-term follow-up of 13 patients with Shwachman's syndrome diagnosed in infancy focusing, in particular, on modifications of the exocrine pancreatic function over time. METHODS: Exocrine pancreatic function was evaluated by duodenal intubation followed by a pancreatic stimulation test. Nutritional, biochemical, hematologic, radiologic, and psychological evaluations were performed at various intervals. Six patients were included in long-term follow-up evaluation. RESULTS: At diagnosis, growth retardation was present in all patients, and all subjects showed pancreatic insufficiency. Hematologic features (intermittent neutropenia, anemia and thrombocytopenia), respiratory infection during the first years of life, and skeletal abnormalities were also frequently observed. Other associated features at diagnosis included hepatic involvement and occasional renal dysfunction. In the six patients followed up, a significant growth improvement was observed. In five of them the pancreatic stimulation test showed values of lipase within reference range outputs, whereas fat balance or fecal fat losses were normal in all but one subject. Of seven subjects assessed by psychological evaluation, IQ test results were markedly abnormal in one and bordered on abnormality in the others. CONCLUSIONS: The present data on Shwachman's syndrome diagnosed in infancy underline the possibility of improvement or normalization of exocrine pancreatic function with age, suggesting the need for periodic checks on pancreatic activity in these subjects. It also indicates the possibility of diagnosis of this syndrome in the absence of pancreatic insufficiency; decreasing frequency of infections over time; and the usefulness of early neuropsychological evaluation. PMID- 10467991 TI - Macronutrient and energy intakes in young children fed milk products containing different quantities and qualities of fat and protein. AB - BACKGROUND: Milk is the major source of protein and saturated fats in the diet after infancy. In the present study, the effects of different fat and protein quantity as well as fat quality in milk and dairy products on nutrient intake and growth in young children were determined. METHODS: Thirty-eight healthy children were randomly assigned to one of four feeding groups at 12 months of age: 1) Low fat milk (1.0 g fat/dl, 3.3 g protein/dl); 2) standard-fat milk (3.5 g fat/dl, 3.3 g protein/dl); 3) partially vegetable fat and protein-reduced milk (3.5 g fat/dl, 50% vegetable; 2.2 g protein/dl); and 4) full-vegetable-fat milk (3.5 g fat/dl, 100% vegetable; 3.0 g protein/dl). Nutrient intake and growth were measured at 12, 15, and 18 months. RESULTS: The protein intake was significantly reduced with the protein-reduced milks. The intake of saturated fat was significantly lower with low-fat milk (11% of energy) or milks containing vegetable fat (13%) than in standard-fat milk (19.6%). However, the total fat intake was below 30% of energy in low-fat milk, whereas the total fat intake remained more than 30% in the other groups. Energy intake and growth were similar in all groups. CONCLUSIONS: A modified milk with reduced protein content for young children results in protein intakes closer to recommendations. A modified milk with either 50% or 100% vegetable fat and dairy products with low milk fat and high vegetable fat content results in intakes of total and saturated fats closer to international recommendations, compared with standard or low-fat milk and dairy products. PMID- 10467992 TI - Gluten-free diet decreases urinary peptide levels in children with celiac disease. AB - BACKGROUND: Increased urine secretion of peptides has been found in celiac disease, probably resulting from increased intestinal uptake of peptides caused by damage to the small gut mucosa. METHODS: High-performance liquid chromatography of low-molecular-weight peptides in the urine was performed over 6 months, before and after a gluten-free diet was instituted in children who clinically improved while consuming the diet. RESULTS: A significant decrease of peptide levels was observed in children consuming the gluten-free diet. Certain peptide peaks thought to be gluten related decreased the most after the patients began the diet. CONCLUSIONS: Because the peptides decrease in patients consuming a gluten-free diet, it is reasonable to conclude that such peptides have a mostly dietary origin. PMID- 10467993 TI - Celiac disease in relation to immunologic serum markers, trace elements, and HLA DR and DQ antigens in Swedish children with Down syndrome. AB - BACKGROUND: An association between Down syndrome and celiac disease has been reported. This study was conducted to determine the association between childhood celiac disease and Down syndrome in the county of Uppsala, Sweden. METHODS: All 76 children with Down syndrome (1-18 years) were screened for the occurrence of anti-gliadin antibodies (AGA) and anti-endomysium antibodies (EMA). Twelve children with suspected celiac disease were investigated further. RESULTS: Increased levels of both IgA and IgG AGA were found in 26% of the children and of EMA in and 5 of 76. Celiac disease was diagnosed in at least three of the children (3.9%; 95% confidence interval 0%-8.3%), and it could have been present in as many as eight. Three of the five EMA-positive children with suspected celiac disease had the HLA phenotype DR3, DQ2. CONCLUSIONS: The results show that determination of EMA is more useful as a screening test for celiac disease and for follow-up than is AGA in children with Down syndrome. The present study also confirms that celiac disease is overrepresented among Swedish children with Down syndrome and that celiac disease should be considered in all persons with Down syndrome. PMID- 10467994 TI - Effect of sequential erythromycin and octreotide on antroduodenal manometry. AB - BACKGROUND: In earlier studies, erythromycin stimulated but octreotide inhibited gastric antral contractions, as each drug induced phase 3-like episodes. METHODS: To assess the effect of erythromycin pretreatment on octreotide-induced changes in antroduodenal motility, 16 patients were studied (mean age, 8.7 +/- 1.5 years, 8 male): 6 with severe gastroesophageal reflux, 4 with cyclic vomiting, 3 with gastroparesis, 2 with chronic intestinal pseudo-obstruction, and 1 with Crohn's disease and unexplained nausea and vomiting. After recording fasting antroduodenal motility for 3 hours, 1 mg/kg intravenous erythromycin was administered over 30 minutes. Sixty minutes after the erythromycin infusion, 0.5 microg/kg subcutaneous octreotide was administered, followed 1 hour later by a meal. RESULTS: Phase 3 occurred spontaneously in 10 patients and after erythromycin in 12 patients. When administered after erythromycin, octreotide immediately induced phase 3s contractions in 15 patients, beginning in the antrum. In 7 children, some of the octreotide-induced phase 3s did not propagate. After the meal, antral contractions continued in all patients. The fed pattern was replaced in 14 patients by alternating phase 3 and phase 1 activities. CONCLUSIONS: Pretreatment with erythromycin prevented octreotide-induced inhibition of antral contractions. Inhibition of antral contractions by octreotide may be mediated through either a direct or indirect suppression of motilin release, because antral contractions persist after pretreatment with the motilin receptor agonist erythromycin. PMID- 10467995 TI - Normalizing results of 13C-urea breath testing for CO2 production rates in children. AB - BACKGROUND: The 13C-urea breath test detects the presence of Helicobacter pylori from an enrichment of breath 13CO2, which, in turn, is critically dependent on the amount of dilution by endogenous CO2 production. The production of CO2 differs according to age (adults > children), sex (male > female) weight, and height. The cutoff value of 2.4 delta%(delta over baseline, DOB) for the 13C-urea breath test, defined in adults, does not take into account actual CO2 production. Therefore, this cutoff value (2.4 delta%) may or may not be appropriate for children. The purpose of this study was to determine a cutoff value that would provide accurate results in pediatric patients, independent of their differences in anthropometric parameters. METHODS: Estimates of CO2 production were combined with DOB values to calculate the host-dependent urea hydrolysis rate. RESULTS: Calculated as urea hydrolysis rate, the cutoff range for adults was 10.4 to 10.9 microg/min. Individual ranges were concentric (men, 9.6-10.9 microg/min; women, 8.5-12.2 microg/min). Results in studies of 312 children show that a urea hydrolysis rate of more than 10 m microg/min may also be appropriate to predict H. pylori infection. CONCLUSION: Calculating 13C-urea breath test values as urea hydrolysis rate removes the effect of individual anthropometric differences on test outcome and provides a single cutoff value for pediatric patients of all ages. PMID- 10467996 TI - Testing for serum IgG antibodies to Helicobacter pylori cytotoxin-associated protein detects children with higher grades of gastric inflammation. AB - BACKGROUND: Little information is available about the relationships between Helicobacter pylori cytotoxin-associated protein (CagA) and clinicopathologic features in children. The purpose of this study was to test whether determining serum IgG antibodies to CagA is a useful tool for detecting more severe disease. METHODS: One hundred twenty-seven consecutive children (age range, 0.75-17.8 years; median, 9.4 years) referred for gastroscopy were included in the study. Antral and corpus biopsies were taken for gastric histology and H. pylori detection. Major symptoms and endoscopic findings were recorded. A serum sample was drawn from each child and assayed for IgG antibodies CagA by a commercial enzyme-linked immunosorbent assay. RESULTS: Sixty-three (50%) children had no evidence of H. pylori infection, 28 (22%) were H. pylori positive/CagA positive, and 36 (28%) were H. pylori positive/CagA negative. There were no differences in clinical diagnosis and occurrence of any predominant symptom according to H. pylori and CagA status. Findings of antral nodularity were more frequent (p = 0.003) in H. pylori-positive/CagA-positive children than in H. pylori positive/CagA-negative children. The gastritis score was significantly higher in H. pylori-positive/CagA-positive children than in H. pylori-positive/CagA negative children (5.7 +/- 1.9 vs. 3.8 +/- 1.6, respectively; p = 0.0003), either in the antral (p = 0.0002) or in the corpus (p = 0.001) mucosa. Inflammation (p = 0.0001) and activity (p = 0.0001) scores were both higher in H. pylori positive/CagA-positive children than in H. pylori-positive/CagA-negative children, but the H. pylori density score was not significantly different (p = NS). In no case was normal gastric mucosa found in H. pylori-positive/ CagA positive children. Lymphocytic gastritis (p = 0.0008) and lymphoid follicles (p = 0.000003) were a more frequent finding in H. pylori-positive children than in H. pylori negative children, irrespective of CagA status. CONCLUSION: Testing for serum IgG to CagA detects higher grades of gastric inflammation among children with H. pylori infection. It may be useful in targeting H. pylori-positive/ CagA positive children for antimicrobial therapy while reducing the need for endoscopy and gastric biopsy. PMID- 10467997 TI - Differing patterns of transforming growth factor-beta expression in normal intestinal mucosa and in active celiac disease. AB - BACKGROUND: Growth-inhibitory autocrine polypeptides such as transforming growth factor (TGF)-beta may play a role in the control of normal epithelial cell proliferation and differentiation. In addition, TGF-beta has a central role in extracellular matrix homeostasis and regulates the immune response at the local level. In this study immunohistochemistry was used to examine the pattern of TGF beta protein distribution and quantitative reverse transcription-polymerase chain reaction (RT-PCR) to determine levels of TGF-beta messenger RNA expression in normal intestinal mucosa and in the flat mucosa of children with celiac disease. METHODS: Small intestinal biopsies were performed in children with active celiac disease and in histologically normal control subjects. Frozen sections were single stained using an anti-TGF-beta monoclonal antibody and were double stained for TGF-beta and T cell, macrophages, and the activation marker CD25. Total RNA was extracted from frozen specimens and competitive quantitative RT-PCR performed for TGF-beta mRNA using internal synthetic standard RNA. RESULTS: In normal intestinal mucosa, by immunohistochemistry, TGF-beta expression was most prominent in the villous tip epithelium, whereas in the lamina propria, weak immunoreactivity was present. The celiac mucosa showed weak and patchy epithelial TGF-beta immunoreactivity. In contrast, an intense staining positivity was present in the lamina propria localized mostly in the subepithelial region where T cells, macrophages, and CD25+ cells were detected by double staining. By quantitative RT-PCR, levels of TGF-beta mRNA transcripts appeared to be increased in celiac intestinal mucosa compared with that in control subjects, although the difference did not reach statistical significance. CONCLUSIONS: These observations suggest that TGF-beta expression is associated with differentiated enterocyte function. In celiac disease the lower TGF-beta epithelial cell expression could be a consequence of the preponderance of a less differentiated epithelial cell phenotype also present in the surface epithelium. In contrast, the prominent TGF-beta positivity of the subepithelial lamina propria suggests an association with the local immune and inflammatory response, as well as a potential role of these peptides in mesenchymal-epithelial cell interaction. PMID- 10467998 TI - Serum levels of hepatitis C virus RNA in infants and children with chronic hepatitis C. AB - BACKGROUND: The role of serum hepatitis C virus (HCV) load in infectivity, disease activity, and response to interferon treatment has been investigated in adults, and controversial results have been obtained. Little is known about HCV load in infants and children with HCV infection. PURPOSE: To investigate the relation between HCV load in serum and features of associated liver disease in infants and children with HCV infection. METHODS: Hepatitis C viral load was investigated in serial samples in 43 children with chronic HCV infection, including 32 patients aged 4 to 16 years infected by different routes and 11 vertically infected infants observed prospectively since birth. RESULTS: Overall viremia ranged between 2.7 and 6.9 log copies/ml (median, 5.56 log/ml) and fluctuated slightly during the follow-up. Median HCV RNA levels did not significantly differ among infants, children, and adolescents. Viral load was also independent of sex, route of infection, clinical manifestation, alanine aminotransferase levels, and liver histology. All 11 perinatally infected children became chronic HCV carriers, whatever their initial viral load; retrospective testing of sera taken in the first day of life in three infants showed high viremia levels. CONCLUSIONS: Viremia levels observed in children were similar to those reported in adults, were independent of age, biochemical activity of liver disease, and chronicity of infection. They were also relatively stable, suggesting that serial measurement of viral load is useless in untreated infants and children. The detection of viremia at birth in children in whom chronic hepatitis developed later suggests the possibility of in utero infection. PMID- 10467999 TI - Evaluation of a long-chain polyunsaturated fatty acid supplemented formula on growth, tolerance, and plasma lipids in preterm infants up to 48 weeks postconceptional age. AB - BACKGROUND: The last trimester of pregnancy is a period of rapid accretion of long-chain polyunsaturated fatty acids, both in the central nervous system and the body as a whole. Human milk contains these fatty acids, whereas some preterm infant formulas do not. Infants fed formulas without these fatty acids have lower plasma and erythrocyte concentrations than infants fed human milk. Preclinical and clinical studies have demonstrated that single-cell sources (algal and fungal) of long-chain polyunsaturated fatty acids are bioavailable. A balanced addition of fatty acids from these oils to preterm formula results in blood fatty acid concentrations in low birth weight infants comparable to those of infants fed human milk. METHODS: In the present study the growth, acceptance (overall incidence of discontinuation, reasons for discontinuation, overall incidence and type of individual adverse events), and plasma fatty acid concentrations were compared in three groups of infants fed a long-chain polyunsaturated fatty acid supplemented preterm infant formula, an unsupplemented control formula, or human milk. The study was prospective, double-blind (formula groups only), and randomized (formula groups only). Two hundred eighty-eight infants were enrolled (supplemented formula group, n = 77; control formula group, n = 78; human milk group, n = 133). RESULTS: Anthropometric measurements at enrollment, at first day of full oral feeding, and at both 40 and 48 weeks postconceptional age did not differ between the formula groups, whereas the human milk-fed group initially grew at a lower rate. The incidence of severe adverse events was rare and not significantly different between formula groups. The groups fed either human milk or supplemented formula had long-chain polyunsaturated fatty acid concentrations higher than those in the control formula group. CONCLUSIONS: The results of this study demonstrate the safety and efficacy of a preterm formula supplemented with long-chain polyunsaturated fatty acids from single-cell oils. PMID- 10468000 TI - 13C and H2 breath tests to study extent and site of starch digestion in children with cystic fibrosis. AB - BACKGROUND: Starch is an important source of energy for children with cystic fibrosis, but little is known about their capacity to digest it. METHODS: A 13C breath test was used to measure starch digestion and oxidation in 16 children with cystic fibrosis (median [range] age, 7.9 [4-15] years; 7 girls, 9 boys) and 5 normal healthy control subjects (median age, 8.3 [7-13] years; 3 girls, 2 boys). A test meal of 13C flour and lactulose was consumed and breath samples were obtained half-hourly thereafter for 6 hours to measure 13C enrichment by isotope ratio mass spectrometry and H2 by electrochemistry. The test was repeated on 10 children with cystic fibrosis when they were taking pancreatic supplements. RESULTS: The median (range) cumulative percentage 13C dose recovery (cPDR), was 35% (18-52%) in control subjects, 18% (9-33%) in children with cystic fibrosis without enzymes, and 29% (22-51%) in those with pancreatic supplements. cPDR differed significantly between healthy control subjects and children with cystic fibrosis without enzymes (p = 0.01) and between children with cystic fibrosis with and without enzymes (p < 0.0001), but there was no difference between control subjects and children with cystic fibrosis taking enzymes (p = 0.5). Eight children with cystic fibrosis had a cPDR within control range, and in six there was a second peak in 13CO2 enrichment coincident with an increase in H2. CONCLUSIONS: Starch digestion and oxidation are diminished in children with cystic fibrosis, but pancreatic enzymes restored them to near normal levels. A second peak in 13CO2 enrichment, suggestive of colonic starch fermentation was absent in healthy children, but present in some children with cystic fibrosis and abolished by pancreatic enzymes. PMID- 10468001 TI - Individualized protein fortification of human milk for preterm infants: comparison of ultrafiltrated human milk protein and a bovine whey fortifier. AB - BACKGROUND: To improve the nutritional management of pre-term infants, a new individualized human milk fortification system based on presupplementation milk protein analyses was evaluated. METHODS: In an open, prospective, randomized multicenter study, 32 healthy preterm infants (birth weights, 920-1750 g) were enrolled at a mean of 21 days of age (range, 9-36 days) when tolerating exclusive enteral feedings of 150 ml/kg per day. All infants were fed human milk and were randomly allocated to fortification with a bovine whey protein fortifier (n = 16) or ultrafiltrated human milk protein (n = 16). All human milk was analyzed for protein content before fortification with the goal of a daily protein intake of 3.5 g/kg. During the study period (mean, 24 days) daily aliquots of the fortified milk were obtained for subsequent analyses of the protein content. RESULTS: Both fortifiers were well tolerated, and growth gain in weight, length, and head circumference, as well as final preprandial concentrations of serum urea, transthyretin, transferrin, and albumin were similar in both groups. The ultimate estimated protein intake was equivalent in both groups (mean 3.1+/-0.1 g/kg per day). Serum amino acid profiles were similar in both feeding groups, except for threonine (significantly higher in the bovine fortifier group) and proline and ornithine (significantly higher in the human milk protein group). CONCLUSIONS: Protein analyses of the milk before individual fortification provides a new tool for an individualized feeding system of the preterm infant. The bovine whey protein fortifier attained biochemical and growth results similar to those found in infants fed human milk protein exclusively with the corresponding protein intakes. PMID- 10468002 TI - Linear growth and skeletal maturation in subjects with treated celiac disease. AB - BACKGROUND: The aim of this study was to report on the long-term growth and development in a group of treated patients with celiac disease. METHODS: The study includes 26 patients (11 boys and 15 girls) with typical celiac disease who were younger than 2.5 at diagnosis and were followed by means of a growth longitudinal monitoring from the introduction of a gluten-free diet (mean age, 1.7 +/- 0.5 years) until adulthood, over a median period of 15.3 years. Growth indicators used were: height, skeletal age, weight and BMI. RESULTS: At the time of admission, the patients had a general tendency to short stature, underweight and retarded skeletal maturation. They did not catch up completely in height and skeletal age after a dietary treatment period of 3 years. Most of them were seen to be slightly below average height for age during childhood and adolescence with skeletal maturity retardation, even if a fairly large interindividual variation of height profiles was evident. CONCLUSIONS: Notwithstanding the early treatment, the careful follow-up, and the good adhesion to the dietary rules of the patients under study, slight negative effects of the disease on growth were not avoided. PMID- 10468003 TI - Fruit juice consumption and the prevalence of obesity and short stature in german preschool children: results of the DONALD Study. Dortmund Nutritional and Anthropometrical Longitudinally Designed. AB - BACKGROUND: In recent years, a possible association between excessive consumption of fruit juice (> or =12 fl oz per day) and short stature and/or obesity has been discussed. The association among the consumption of fruit juice, anthropometric indices, and the overall diet was examined during a 3-year period in a sample of healthy preschool children participating in the Dortmund Nutritional and Anthropometrical Longitudinally Designed (DONALD) Study. METHODS: Two hundred five children were examined annually at the ages of 3, 4, and 5 years. Dietary intake was calculated from 3-day weighed diet records. Height was measured using a stadiometer. Weight was measured using an electronic scale. RESULTS: Five children consumed excessive fruit juice continually in all three records, 10 children in two records, and 23 children in one record. None of the five children with repeatedly excessive fruit juice consumption was obese or short. Growth velocity, body mass index, and height standard deviation score were not correlated with fruit juice consumption. Consumption of fruit juice was inversely correlated with the consumption of all other beverages and the total consumption of all other food. The intake of protein, fat, and carbohydrates of children consuming excessive fruit juice was closer to the international dietary preventive guidelines than the intake of children consuming low amounts of fruit juice. CONCLUSIONS: In the study sample, even repeatedly excessive fruit juice consumption had no influence on anthropometric indices. The results do not justify a general warning or a general promotion regarding high fruit juice consumption in preschool children's diets. PMID- 10468004 TI - Clinical quiz. 5-ASA hypersensitivity lung disease. PMID- 10468005 TI - Endoscopic retrograde cholangiopancreatography and endobiliary stenting for the treatment of a bile leak in a child. PMID- 10468006 TI - Gastric leiomyosarcoma presenting as severe iron-deficiency anaemia. PMID- 10468007 TI - Hemorrhagic ascites in a child with Henoch-Schonlein purpura. PMID- 10468008 TI - Downhill esophageal varices. PMID- 10468009 TI - Pancreatic ascites with pancreatic stone formation in a child. PMID- 10468010 TI - Effect of rhesus rotavirus vaccine on enteric adenovirus--associated diarrhea in children. PMID- 10468012 TI - News from the National Institute of General Medical Sciences (NIGMS). Communicating with the public--it's in your best interest. PMID- 10468011 TI - Flunitrazepam and cisapride in premedication for peroral small intestinal biopsy in children and adolescents. PMID- 10468013 TI - An interview with a distinguished pharmaceutical scientist. PMID- 10468014 TI - Fluorescence recovery after photobleaching: a versatile tool for mobility and interaction measurements in pharmaceutical research. AB - This review introduces the basics of fluorescence recovery after photobleaching (FRAP) from a theoretical and an instrumentational approach. The most interesting and innovative applications with a pharmaceutical point of view are briefly discussed and possible future applications are suggested. These future applications include research on the mobility of macromolecular drugs in macro- or microscopic pharmaceutical dosage forms, mobility, and binding of antitumor drugs in tumor tissue, intracellular trafficking of gene complexes and mobility of drugs in membranes prior to transmembrane penetration. The paper is also intended to be an introductory guideline to those who would like to get involved in FRAP related experimental techniques. Therefore, comprehensive details on different setups and data analysis are given, as well as a brief outline of the problems that may be encountered when performing FRAP. Overall, this review shows the great potential of FRAP in pharmaceutical research. This is complemented by our own results illustrating the possibility of performing FRAP in microscopic dosage forms (microspheres) using a high resolution variant of FRAP. PMID- 10468015 TI - Design of potent phosphorothioate antisense oligonucleotides directed to human interleukin 10 gene product and their evaluation of antisense activity in U937 cells. AB - PURPOSE: The two objectives of this study were to design potent phosphorothioate antisense oligonucleotides (AS-S-oligos) directed against the human interleukin 10 (IL-10) gene product and to reveal the DNA sequence which best activates antisense effects. METHODS: The design of potent AS-S-oligo was performed by using melting temperature (Tm) value of a DNA/RNA hybrid calculated by the nearest neighbor method and a secondary structure of human IL-10 mRNA suggested by RNA folding algorithms. U937 cells were used to estimate the antisense effect of the AS-S-oligos. RESULTS: Of the eight candidates selected as potent AS-S oligos on the basis of having higher Tm values and favorable secondary structures of the IL-10 mRNA, AS-S-oligos directed against the translated (AS367-S-oligo) and 3'-untranslated (AS637-S-oligo) region of IL-10 mRNA showed the strongest inhibitory effects on IL-10 production and this inhibition was dose- and time dependent. Reverse transcription-polymerase chain reaction (RT-PCR) revealed that the antisense effects of AS-S-oligos originated from a specific reduction of target IL-10 mRNA by hybridization with AS367- and AS637-S-oligos. In addition, these AS-S-oligos did not affect human tumor necrosis factor-alpha (TNF-alpha) production in the cells stimulated by lipopolysaccharide (LPS). Strong positive correlations between the inhibitory effect of AS-S-oligos on the IL-10 production and not only Tm values calculated by nearest neighbor method but also Tm values determined by absorbance versus temperature profiles were demonstrated except for AS25-S-oligo and AS1249-S-oligo. CONCLUSIONS: These findings suggest AS367- and AS637-S-oligos powerfully inhibit IL-10 production in U937 cells via an antisense mechanism. In addition, it is suggested efficiency of AS-S-oligo directed against the sequence of the target gene product can be explained by these Tm values and the proposed secondary structures of the target gene product. PMID- 10468016 TI - Synthesis and in vitro evaluation of aminoacyloxyalkyl esters of 2-(6-methoxy-2 naphthyl)propionic acid as novel naproxen prodrugs for dermal drug delivery. AB - PURPOSE: To synthesize and evaluate various novel aminoacyloxyalkyl esters of naproxen (3a-i) and naproxenoxyalkyl diesters of glutamic and aspartic acids (3j m) as potential dermal prodrugs of naproxen. METHODS: The prodrugs 3a-m were synthesized, and their aqueous solubilities, lipophilicities and hydrolysis rates were determined in a buffered solution and in human serum. The permeation of selected prodrugs across excised postmortem human skin was studied in vitro. RESULTS: The aminoacyloxyalkyl prodrugs showed higher aqueous solubilities and similar lipid solubilities, in terms of octanol-buffer partition coefficients (log Papp) at pH 5.0, when compared with naproxen. At pH 7.4 the prodrugs were significantly more lipophilic than naproxen. Prodrugs 3a-i showed moderate chemical stability in aqueous solutions at pH 5.0 and were rapidly converted to naproxen in human serum (t1/2 = 4-19 min). The selected aminoacyloxyalkyl prodrugs possessed a higher flux across the skin than naproxen, with a maximum enhancement of 3-fold compared to naproxen. Prodrugs 3j-m showed poor aqueous solubility and permeation across the skin. CONCLUSIONS: Combinations of adequate aqueous solubility and lipophilicity of naproxen aminoacyloxyalkyl prodrugs having fast rates of enzymatic hydrolysis resulted in improved dermal delivery of naproxen. PMID- 10468017 TI - Synthesis of a lipophilic prodrug of 9-(2-phosphonylmethoxyethyl)adenine (PMEA) and its incorporation into a hepatocyte-specific lipidic carrier. AB - PURPOSE: 9-(2-Phosphonylmethoxyethyl)adenine (PMEA), a potent inhibitor of Hepatitis B virus replication, is in vivo hardly taken up by parenchymal liver cells (the site of infection). Our aim is to examine whether lactosylated reconstituted HDL (LacNeoHDL), a lipidic particle that is specifically internalized by parenchymal liver cells, is a suitable carrier for the selective delivery of PMEA to this cell type. METHODS: To incorporate PMEA into LacNeoHDL, we synthesized a lipophilic prodrug (PMEA-LO) by coupling PMEA via an acid-labile phosphonamidate bond to lithocholic acid-3alpha-oleate. RESULTS: The yield of the synthesis was 52% ([3H]PMEA-LO: 24%). [3H]PMEA-LO readily incorporated into LacNeoHDL (13 molecules/particle) without affecting the size and net negative charge of the carrier. Further, incubation studies at lysosomal pH showed [3H]PMEA was completely released from the carrier whereas, at neutral pH or in plasma, appreciable release was not observed. CONCLUSIONS: The conjugation of PMEA with lithocholic acid-3alpha-oleate results in a lipophilic prodrug that readily associates with Lac-NeoHDL. The association of the prodrug does not affect the physicochemical properties of the particle, and PMEA is released from the carrier at lysosomal pH. These findings indicate that by using the prodrug approach, LacNeoHDL is a suitable carrier to deliver PMEA to parenchymal liver cells. PMID- 10468018 TI - Solid-state characteristics of amorphous sodium indomethacin relative to its free acid. AB - PURPOSE: Having previously studied the amorphous properties of indomethacin (IN) as a model compound for drugs rendered amorphous during processing, we report on the formation and characterization of its sodium salt in the amorphous state and a comparison between the two systems. METHODS: Sodium indomethacin (SI) was subjected to lyophilization from aqueous solution, rapid precipitation from methanol solution, and dehydration followed by grinding to produce, in each case, a completely amorphous form. The amorphous form of SI was analyzed using DSC, XRD, thermomicroscopy and FTIR. The method of scanning rate dependence of the glass transition temperature, Tg, was used to estimate the fragility of the SI system. Enthalpy relaxation experiments were carried out to probe the molecular mobility of the SI system below Tg. RESULTS: The amorphous form of SI formed by different methods had a Tg equal to 121 degrees C at a scanning rate of 20 degrees C/min. This compares with a Tg for indomethacin of 45 degrees C. Estimation of fragility by the scanning rate dependence of Tg indicates no significant differences in fragility between ionized and unionized forms. Enthalpy relaxation measurements reveal very similar relaxation patterns between the two systems at the same degree of supercooling relative to their respective Tg values. CONCLUSIONS: The amorphous form of SI made by various methods has a Tg that is about 75 degrees C greater than that of IN, most likely because of the greater density and hence lower free volume of SI. Yet, the change of molecular mobility as a function of temperature relative to Tg is not very different between the ionized and unionized systems. PMID- 10468019 TI - Compaction properties of composite particles consisting of lactose with sodium alginate prepared by spray-drying. AB - PURPOSE: Composite particles of lactose with a small amount of sodium alginate were prepared by spray-drying (SD) in an effort to improve the compactibility of the polymer for direct compression. The compaction behavior of the SD composite particles with a range of polymer contents was investigated. METHODS: Composite particles were prepared by spray-drying an aqueous solution of lactose and sodium alginate at various formulating ratios. Improvement in the compactibility of the composite particles was evaluated by measuring the tablet tensile strength, porosity-applied pressure profiles, stress relaxation, elastic recovery of the compressed powder, and surface properties of the tablets by scanning electron microscopy. RESULTS: The tensile strength of compacts formed from the SD composite particles containing sodium alginate (< or = 10 wt%) was as high as that of spray-dried amorphous lactose. The improved compaction was attributed to the higher relaxation pressure and lower elastic recovery of the composite particles compared with alpha-lactose monohydrate. However, increasing the sodium alginate content of the SD composite particles above 10 wt% led to a marked reduction in the tensile strength of the resultant tablets. Scanning electron micrographs revealed that composite particles with a good compactibility fused totally in the tablets while composite particles containing 15% or more sodium alginate retained their shape, even after compression. The presence of sodium alginate layered uniformly on the surface of the particles and the increase in the glass transition temperature of the particles, possibly due to interpolation of sodium alginate are responsible for the reduction in the fusion property of the composite particles on compression. CONCLUSIONS: Although increasing the sodium alginate content of SD composite particles led to an increase in their plastic deformation, fusion on compression was prevented by the presence of sodium alginate. The reduced compactibility of SD composite particles with an excess amount of sodium alginate was attributed to reduced cohesion and fusion of the particles during compression. PMID- 10468020 TI - Comparison of CYP2D6 content and metoprolol oxidation between microsomes isolated from human livers and small intestines. AB - PURPOSE: To assess the role of intestinal CYP2D6 in oral first-pass drug clearance by comparing the enzyme content and catalytic activity of a prototype CYP2D6 substrate, metoprolol, between microsomes prepared from human intestinal mucosa and from human livers. METHODS: Microsomes were prepared from a panel of 31 human livers and 19 human intestinal jejunal mucosa. Microsomes were also obtained from the jejunum, duodenum and ileum of four other human intestines to assess regional distribution of intestinal CYP2D6. CYP2D6 content (pmole/mg microsomal protein) was determined by Western blot. CYP2D6 activity was measured by alpha-hydroxylation and O-demethylation of metoprolol. RESULTS: Kinetic studies with microsomes from select livers (n = 6) and jejunal mucosa (n = 5) yielded K(M) estimates of 26 +/- 9 microM and 44 +/- 17 microM, respectively. The mean Vmax (per mg protein) for total formation of alpha-OH-M and ODM was 14-fold higher for the liver microsomes compared to the jejunal microsomes. Comparisons across intestinal regions showed that CYP2D6 protein content and catalytic activity were in the order ofjejunum > duodenum > ileum. Excluding the poor metabolizer genotype donors, CYP2D6 content varied 13- and 100-fold across the panels of human livers (n = 31) and jejunal mucosa (n = 19), respectively. Metoprolol alpha-hydroxylation activity and CYP2D6 content were highly correlated in the liver microsomes (r = 0.84, p < 0.001) and jejunal microsomes (r = 0.75, p < 0.05). Using the well-stirred model, the mean microsomal intrinsic clearance (i.e., Vmax/K(M)) for the livers and jejunum were scaled to predict their respective in vivo organ intrinsic clearance and first-pass extraction ratio. Hepatic and intestinal first-pass extractions of metoprolol were predicted to be 48% and 0.85%, respectively. CONCLUSIONS: A much lower abundance and activity of CYP2D6 are present in human intestinal mucosa than in human liver. Intestinal mucosal metabolism contributes minimally to the first-pass effect of orally administered CYP2D6 substrates, unless they have exceptionally high microsomal intrinsic clearances and/or long residence time in the intestinal epithelium. PMID- 10468021 TI - Role of P-glycoprotein on the CNS disposition of amprenavir (141W94), an HIV protease inhibitor. AB - PURPOSE: To determine the role of P-glycoprotein (Pgp) on the CNS penetration of the HIV protease inhibitor (PI) amprenavir (141W94) and to test the hypothesis that co-administration of a second HIV PI (ritonavir) could enhance amprenavir's brain penetration in vivo. METHODS: Pgp-mediated efflux was investigated in vitro with Caco-2 cells and in vivo by whole-body autoradiography (WBA). "Genetic" mdr1a/1b double knockout mice, "chemical" Pgp knockout mice generated by administration of the Pgp inhibitor GF120918, and mice pretreated with ritonavir were used in WBA studies to investigate the effects of Pgp modulation on the CNS penetration of amprenavir. RESULTS: Amprenavir, indinavir, ritonavir, and saquinavir had 2- to 23-fold higher transport rates from the basolateral to apical direction than from the apical to basolateral direction across Caco-2 monolayers. Incubation with GF120918 negated this difference, suggesting that the efflux was Pgp-mediated. WBA studies demonstrated a 13- and 27-fold increase in the brain and a 3.3-fold increase in the CSF concentrations of amprenavir in mice pretreated with GF120918 and in mdr1a/1b double knockout mice. In contrast, pretreatment with ritonavir did not alter the CNS exposure of amprenavir. CONCLUSIONS: These results provide evidence that amprenavir and other HIV PIs are Pgp substrates and that co-administration of a specific Pgp inhibitor will enhance amprenavir's CNS penetration in vivo. These results will have an important therapeutic impact in the treatment of AIDS dementia. PMID- 10468022 TI - P-glycoprotein-dependent disposition kinetics of tacrolimus: studies in mdr1a knockout mice. AB - PURPOSE: This study was performed to evaluate the involvement of P-glycoprotein in disposition kinetics of tacrolimus (FK506), a substrate of P-glycoprotein, in the body. METHODS: The blood and tissue concentrations of FK506 after i.v. or p.o. administration (2 mg/kg) to normal and mdr1a knockout mice were measured by competitive enzyme immunoassay. RESULTS: The blood concentrations in knockout mice were significantly higher than those in normal mice. The value of the total clearance (CLtot) for knockout mice (19.3 mL/min/kg) was about 1/3 of that for normal mice (55.8 mL/min/kg)(P < 0.001), although there was no significant difference in the distribution volume at the steady-state (Vd(ss)) (about 4.6 L/kg) between both types of mice. FK506 rapidly penetrated the blood-brain barrier and the brain concentration reached a maximum, which was about 10 times higher in knockout mice than in normal mice, 1 hr after administration. The brain concentration in normal mice thereafter decreased slowly, whereas in knockout mice, an extremely high concentration was maintained for 24 hr. CONCLUSIONS: The pharmacokinetic behavior of FK506 in the tissue distribution is related with the function of P-glycoprotein encoded by the mdrla gene. The brain distribution of FK506 is dominated by the P-glycoprotein-mediated drug efflux and presumably also by the binding to FK-binding proteins (immunophilins) in the brain. PMID- 10468023 TI - Pharmacokinetics of methotrexate in the extracellular fluid of brain C6-glioma after intravenous infusion in rats. AB - PURPOSE: Establishment of the pharmacokinetic profile of methotrexate (MTX) in the extracellular fluid (ECF) of a brain C6-glioma in rats. METHODS: Serial collection of plasma samples and ECF dialysates after i.v. infusion of MTX (50 or 100 mg/kg) for 4 h. HPLC assay. RESULTS: Histological studies revealed the presence of inflammation, edema, necrosis, and hemorrhage in most animals. In vivo recovery (reverse dialysis) was 10.8 +/- 5.3%. MTX concentrations in tumor ECF represented about 1-2% of the plasma concentrations. Rapid equilibration between MTX levels in brain tumor ECF and plasma. ECF concentrations almost reached steady-state by the end of the infusion (4 h), then decayed in parallel with those in plasma. Doubling of the dose did not modify MTX pharmacokinetic parameters (t1/2alpha, t1/2beta, MRT, fb, Vd, and CL(T)), except for a 1.7-fold increase of AUC(Plasma) and a 3.8-fold increase in AUC(ECF), which resulted in a 2.3-fold increase in penetration (AUC(ECF)/AUC(Plasma)). In spite of an important interindividual variability, a relationship between MTX concentrations in plasma and tumor ECF could be established from mean pharmacokinetic parameters. CONCLUSIONS: High plasma concentrations promote the penetration of MTX into brain tissue. However, free MTX concentrations in tumor ECF remain difficult to predict consistently. PMID- 10468024 TI - Evaluation of the pharmacokinetic features and tissue distribution of the potent nonnucleoside inhibitor of HIV-1 reverse transcriptase, N-[2-(2-fluorophenethyl)] N'-[2-(5-bromopyridyl)]-thiourea (HI-240) with an analytical HPLC method. AB - PURPOSE: The purpose of the present study was to examine the pharmacokinetic features and tissue distribution of N-[2-(2-fluorophenethyl)]-N'-[2-(5 bromopyridyl)]-thiourea (HI-240), a novel non-nucleoside inhibitor of HIV reverse transcriptase with potent anti-viral activity against AZT-sensitive as well as multidrug-resistant HIV-1 strains. METHODS: A sensitive and accurate high performance liquid chromatography (HPLC)-based quantitative detection method was established to measure concentrations of HI-240 in pharmacokinetic studies. The plasma concentration-time data were modeled by using the WinNonlin program to estimate the pharmacokinetic parameter values. RESULTS: HI-240 had an elimination half-life of 78.3 +/- 2.0 min after i.v. administration and 196.8 +/- 3.1 min after i.p. administration. The systemic clearance of HI-240 was 2194 +/- 61 ml/h/kg after i.v. administration and 9339 +/- 1160 ml/h/kg after i.p. administration. Following i.v. injection, HI-240 rapidly distributed to and accumulated in multiple tissues with particularly high accumulation in adipose tissue, adrenal gland, and uterus+ovary. The concentration of HI-240 in brain tissue was comparable to that in the plasma, indicating that HI-240 easily crosses the blood-brain-barrier. Following i.p. injection, HI-240 was rapidly absorbed with a t1/2ka and a tmax values of less than 10 min. Following oral administration, HI-240 was absorbed with a t1/2ka of 4.2 +/- 1.1 min and a tmax of 95.1 +/- 25.1 min. The intraperitoneal bioavailability was estimated at 23.5%, while the oral bioavailability was only 1%. CONCLUSIONS: The HPLC-based accurate and precise analytical detection method and pilot pharmacokinetic studies described herein provide the basis for advanced preclinical pharmacodynamic studies of HI-240. The ability of HI-240 to distribute rapidly and extensively into extravascular compartments and easily cross the blood-brain barrier represent significant pharmacokinetic advantages over AZT. PMID- 10468025 TI - Impact of regional intestinal pH modulation on absorption of peptide drugs: oral absorption studies of salmon calcitonin in beagle dogs. AB - PURPOSE: To investigate the relationship between the modulation of intestinal pH and the oral absorption properties of a model peptide drug, salmon calcitonin (sCT), in conscious beagle dogs. METHODS: Studies were performed to characterize the disintegration of the formulation, intestinal pH changes, and the appearance of the peptide in the blood. Enteric-coated formulations containing sCT and various amounts of citric acid (CA) were tethered to a Heidelberg capsule (HC) and given orally to normal beagle dogs. Blood samples were collected and analyzed by radioimmunoassay (RIA). Intestinal pH was continuously monitored using the Heidelberg pH capsule (HC) system. The integrity of the HC-delivery system tether was verified by fluoroscopy. RESULTS: The intra-individual variation in gastric emptying (GE) of the delivery system was large. There were also large inter individual differences in the disintegration and absorption properties of the various formulations. However, the peak plasma concentrations of sCT were always observed when the intestinal pH declined. The average baseline intestinal pH was 6.1 +/- 0.2 (mean +/- SEM, n = 12). The intestinal pH reduction was 2.6 +/- 0.4 (mean +/- SEM, n = 12, ranged from 0.5 to 4.0 units from baseline). There was a good correlation between the time to reach the trough intestinal pH (t(pH,min)) and time to reach the peak plasma concentration (tconc,max)) of sCT (t(conc,max) = 0.95 x t(pH,min) + 14.1, n = 11, r2 = 0.91). Plasma Cmax and area under the curve (AUC) increased with increasing amounts of CA in the formulations. CONCLUSIONS: The results of these studies demonstrate that the oral absorption properties of a model peptide drug, sCT, can be modulated by changing intestinal pH. sCT is a substrate for the pancreatic serine protease trypsin which has maximal activity at pH 5 to 6. Reducing intestinal pH presumably stabilizes sCT in the GI tract enabling greater absorption of the intact peptide. PMID- 10468027 TI - Membrane-interaction QSAR analysis: application to the estimation of eye irritation by organic compounds. AB - PURPOSE: The purpose of this study was to explore a potential mechanism of eye irritation, and to construct a corresponding general quantitative structure activity relationship (QSAR) model, in terms of diversity of irritant chemical structure, based on the Draize eye irritation ECETOC data set. METHODS: Molecular dynamic simulation (MDS) was used to generate intermolecular membrane-solute interaction properties. These intermolecular properties were combined with intramolecular physicochemical properties and features of the solute (irritant) to construct QSAR models using multi-dimensional linear regression and the Genetic Function Approximation (GFA) algorithm. RESULTS: Significant QSAR models for estimating eye irritation potential were constructed in which solute aqueous solvation free energy and solute-membrane interaction energies are the principle correlation descriptors. These physicochemical descriptors were selected from a trial set of 95 descriptors for 18 structurally diverse compounds fully representative of the ECETOC set of 38 compounds. CONCLUSIONS: Combining intermolecular solute-membrane interaction descriptors with intramolecular solute descriptors yields statistically significant eye irritation QSAR models. The resultant QSAR models support an eye irritation mechanism of the action in which increased aqueous solubility of the irritant and its strength of binding to the membrane both increase eye irritation. PMID- 10468026 TI - Gastropods as an evaluation tool for screening the irritating potency of absorption enhancers and drugs. AB - PURPOSE: The objective of this study was to develop a simple alternative test using naked snails (slugs) for screening the irritating potency of chemicals on mucosal surfaces. METHODS: The effect of various absorption enhancers and two beta-blocking agents on the mucosal tissue was determined from the total protein and lactate dehydrogenase released from the foot mucosa after treatment. Additionally, mucus production and reduction in body weight of the slugs caused by the treatment were measured. RESULTS: According to the effects on the mucosal epithelium of the slugs the following rank order of increasing toxicity was established: PBS, HP-beta-CD (5%), beta-CD (1.8%) and oxprenolol hydrochloride (1%) < DDPC (1%) < STDHF (1%) < BAC (1%), SDC (1%) and propranolol hydrochloride (1%). The results of the present study are in agreement with other studies using the same compounds on other models. CONCLUSIONS: The results of this study indicated the mucosa of slugs can serve as a primary screening tool for the evaluation of chemicals on mucosal surfaces. By simply measuring mucus production and weight loss reliable toxicity information can be obtained. This demonstrates rapid screening tests can be carried out using simple toxicity endpoints. PMID- 10468029 TI - Likelihood-based diagnostics for influential individuals in non-linear mixed effects model selection. AB - PURPOSE: Data from single individuals, or a small group of subjects may influence non-linear mixed effects model selection. Diagnostics routinely applied in model building may identify such individuals, but these methods are not specifically designed for that purpose and are, therefore, not optimal. We describe two likelihood-based diagnostics for identifying individuals that can influence the choice between two competing models. METHODS: One method is based on a jackknife of the raw data on the individual level and refitting the model to each new data set. The second method is a calculation which utilises the contribution each individual make to the objective function values under each of the two models. The two methods were applied to model selection during analysis of a real data set. RESULTS: The agreement between the methods was high. Individuals for whom there was a discrepancy between the methods tended to be those for which neither of the contending models described the data appropriately. Both methods identified individuals that influenced the model selection. CONCLUSIONS: Two objective, specific and quantitative methods for identifying influential individuals in nonlinear mixed effects model selection have been presented. One of the methods doesn't require additional model fitting and is therefore particularly attractive. PMID- 10468028 TI - Stool water content and colonic drug absorption: contrasting effects of lactulose and codeine. AB - PURPOSE: By varying stool water content using lactulose and codeine, we investigated the influence of luminal water content on the absorption of quinine, a transcellular probe, and 51Cr-EDTA, a paracellular probe, from the distal gut. METHODS: Sixteen volunteers entered a three-way cross-over trial in which absorption of probe markers from a timed-release delivery system was determined following treatment with lactulose 20 mls tds (increasing water content), or codeine 30 gms qds (decreasing water content), and compared with control untreated values. Stool water content was assessed by freeze drying stool samples. Site of release was determined by gamma scintigraphy, and absorption was measured by plasma levels and urinary recovery of the marker probes. RESULTS: Lactulose accelerated ascending colon transit (3.7 +/- 0.8 vs 4.5 +/- 1.4 hrs, p < 0.05), increased stool water content (75 +/- 2 vs 71 +/- 2%, p < 0.01), caused greater dispersion of released material (dispersion score 3.4 +/- 0.3 vs 1.8 +/- 0.2, p < 0.01), and enhanced absorption of the transcellular probe quinine (4.66 +/- 0.78 vs 3.02 +/- 0.63%, p < 0.05) compared to control. Conversely codeine slowed ascending colon transit (8.9 +/- 1.8 hrs), reduced stool water content (61 +/- 2 vs 71.2%, p < 0.05), and tended to diminish absorption (2.60 +/- 0.77 vs 3.02 +/- 0.63%, p = 0.20). Within the ascending colon specifically, there was a significant trend for treatments increasing luminal water content to enhance quinine absorption (medians: codeine = 1.2%, [n = 81 < control = 2.3%, [n = 5] < lactulose = 3.2%, [n = 71, p < 0.01). Delivery site also had an important influence on absorption, with more distal release resulting in less absorption in the control arm (medians: small intestine = 4.4% [n = 5] > ascending colon = 2.3% [n = 5] > transverse colon = 1.5% [n = 6], p < 0.005). CONCLUSIONS: Lactulose accelerates transit, increases stool water content, and enhances drug absorption from the distal gut whilst codeine slows transit, decreases stool water content, and tends to diminish absorption, compared to controls. We conclude that water content may be an important determinant in colonic drug absorption. PMID- 10468030 TI - Transcellular and lipophilic complex-enhanced intestinal absorption of human growth hormone. AB - PURPOSE: To evaluate the transcellular mechanism of novel enhancers absorption enhancement of human growth hormone (hGH), by examining the involvement of a P glycoprotein-like efflux system, changes in membrane fluidity, and membrane damage. METHODS: Caco-2 cell monolayers were grown on Snapwell filter supports and placed in a side-by-side diffusion apparatus. Transport in both the apical to basolateral (AP to BL) and basolateral to apical (BL to AP) direction was measured at different temperatures and in the presence of potential inhibitors. Fluorescence anisotropy measurement was used to measure membrane fluidity. The fluorescence anisotropy of DPH- and TMA-DPH-labeled cell suspensions was measured at room temperature. LDH (a measure of cytosolic lactate dehydrogenase) leakage assay was used to evaluate cytotoxicity. RESULTS: The bi-directional transepithelial fluxes of hGH in the presence of these novel enhancers across Caco-2 cells showed marked asymmetry. Average permeability coefficient values obtained in the apical to basolateral (AP to BL) direction were lower than those of the reverse (BL to AP) direction. On the other hand, the fluxes for hGH alone were symmetric. When P-gp-like efflux inhibitors were included in the transport medium, the permeability coefficient value of BL to AP direction was significantly decreased while the transport was increased in the reverse direction in the presence of novel enhancers. In addition, lowering the temperature to 25 degrees C completely eliminated the asymmetry of hGH transport in the presence of novel enhancers. It was also shown by fluorescence anisotropy that these novel enhancers alone only slightly increased membrane fluidity. On the other hand, upon addition of hGH to the novel enhancers, the cell membrane showed a dramatic change as compared to treatment with novel enhancers alone. The results from the LDH assay showed that the novel enhancers and/or hGH did not cause cell damage, at least up to 1 hour, and the damage seen at the 2 hour point is also much lower than other known enhancers. CONCLUSIONS: This study shows that human growth hormone alone cannot be transported across Caco-2 cells, except in small quantities, by passive diffusion, but in the presence of novel enhancers, human growth hormone permeation is substantial. In addition, the asymmetry of transport of the complexed hGH appears to be due to a P-gp-like efflux system. Assuming that the present substrate specificity of the P-gp-like efflux system shows the same preference for hydrophobic molecules as p-gp, the present work also indirectly shows that human growth hormone has become more lipophilic in the presence of these novel enhancers. Furthermore, membrane fluidity data also supports the premise that these novel enhancers interact and stabilize hGH, to make them more hydrophobic and easier to be transported through cell membranes. PMID- 10468031 TI - A novel non-viral vector for DNA delivery based on low molecular weight, branched polyethylenimine: effect of molecular weight on transfection efficiency and cytotoxicity. AB - PURPOSE: Low molecular weight branched polyethylenimine (LMW-PEI) was synthesized and studied as a DNA carrier for gene delivery with regard to physico-chemical properties, cytotoxicity, and transfection efficiency. METHODS: The architecture of LMW-PEI, synthesized by acid catalyzed ring-opening polymerization of aziridine was characterized by size exclusion chromatography in combination with laser light scattering and 13C-NMR-spectroscopy. In vitro cytotoxic effects were quantified by LDH and MTT assay and visualized by transmission electron microscopy. The potential for transgene expression was monitored in ECV304 cells using luciferase driven by a SV40 promotor as reporter gene system. RESULTS: LMW PEI (Mw 11'900 D) with a low degree of branching was synthesized as a DNA carrier for gene delivery. In contrast to high molecular weight polyethylenimines (HMW PEI; Mw 1'616'000 D), the polymer described here showed a different degree of branching and was less cytotoxic in a broad range of concentrations. As demonstrated by transmission electron microscopy the LMW-PEI formed only small aggregates which were efficiently taken up by different cells in the presence of serum, most likely by an endocytic pathway. LMW-PEI yielded transfection efficiencies measured via expression of the reporter gene luciferase which were up to two orders of magnitude higher than those obtained with HMW-PEI. The reporter gene expression was concentration dependent, but in contrast to lipofection independent of serum addition. CONCLUSIONS: The LMW-PEI described here is a new, highly efficient, and non-cytotoxic vector with a favorable efficiency/toxicity profile for gene therapeutic applications. PMID- 10468032 TI - Organic cation transport in rabbit alveolar epithelial cell monolayers. AB - PURPOSE: To characterize organic cation (OC) transport in primary cultured rabbit alveolar epithelial cell monolayers, using [14C]-guanidine as a model substrate. METHODS: Type II alveolar epithelial cells from the rabbit lung were isolated by elastase digestion and cultured on permeable filters precoated with fibronectin and collagen. Uptake and transport studies of [14C]-guanidine were conducted in cell monolayers of 5 to 6 days in culture. RESULTS: The cultured alveolar epithelial cell monolayers exhibited the characteristics of a tight barrier. [14C]-Guanidine uptake was temperature dependent, saturable, and inhibited by OC compounds such as amiloride, cimetidine, clonidine, procainamide, propranolol, tetraethylammonium, and verapamil. Apical guanidine uptake (Km = 129 +/- 41 microM, Vmax = 718 +/- 72 pmol/mg protein/5 min) was kinetically different from basolateral uptake (Km = 580 +/- 125 microM, Vmax = 1,600 +/- 160 pmol/mg protein/5 min). [14C]-Guanidine transport across the alveolar epithelial cell monolayer in the apical to basolateral direction revealed a permeability coefficient (Papp) of (7.3 +/- 0.4) x 10(-7) cm/sec, about seven times higher than that for the paracellular marker [14C]-mannitol. CONCLUSIONS: Our findings are consistent with the existence of carrier-mediated OC transport in cultured rabbit alveolar epithelial cells. PMID- 10468033 TI - Chemical uptake into human stratum corneum in vivo from volatile and non-volatile solvents. AB - PURPOSE: Simple, safe and quick in vivo methods for estimating chemical uptake into the stratum corneum (SC) from volatile and non-volatile solvents are invaluable to health risk assessors. This study compares the human in vivo SC uptake of a model compound (4-cyanophenol) from water and acetone using quantitative attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy. METHODS: Small areas on the ventral forearms of human volunteers were treated with 4-cyanophenol (CP) dissolved either in water or acetone. After the skin was cleansed of remaining surface CP, SC samples were taken by a standard tape-stripping method. CP concentration profiles across the SC were quantitated by direct measurement of the permeant on the individual tape-strips using ATR-FTIR. RESULTS: Increasing the duration of exposure to CP aqueous solutions resulted in increasing CP uptake into the SC; the kinetics of uptake correlated well with predictive diffusion equations. Increasing the 'dose' of CP in acetone also resulted in increasing uptake into the SC, but uptake eventually plateaued at a maximum level. The amount of CP taken up into the SC from acetone was 2 to 8-fold greater than that from water following similar short-time exposures. CONCLUSIONS: These safe, simple experimental methods provide practical and predictive assessments of chemical uptake into human SC in vivo. PMID- 10468034 TI - Why does PEG 400 co-encapsulation improve NGF stability and release from PLGA biodegradable microspheres? AB - PURPOSE: The aim of this work was to understand the mechanism by which co encapsulated PEG 400 improved the stability of NGF and allowed a continuous release from PLGA 37.5/25 microspheres. METHODS: Microparticles were prepared according to the double emulsion method. PEG 400 was added with NGF in the internal aqueous phase (PEG/PLGA ratio 1/1 and 1.8/1). Its effect was investigated through interfacial tension studies. Protein stability was assessed by ELISA. RESULTS: A novel application of PEG in protein stabilization during encapsulation was evidenced by adsorption kinetics studies. PEG 400 limited the penetration of NGF in the interfacial film of the primary emulsion. Consequently, it stabilized the NGF by reducing the contact with the organic phase. In addition, it avoided the NGF release profile to level off by limiting the irreversible NGF anchorage in the polymer layers. On the other hand, the amount of active NGF released in the early stages was increased. During microparticle preparation, NaCl could be added in the external aqueous phase to modify the structure of microparticles. This allowed to reduce the initial release rate without affecting the protein stability always encountered in the absence of PEG. CONCLUSIONS: PEG 400 appeared of major interest to achieve a continuous delivery of NGF over seven weeks from biodegradable microparticles prepared by the double emulsion technique. PMID- 10468035 TI - Controlled DNA delivery systems. AB - PURPOSE: Genes are of increasing interest as pharmaceuticals, but current methods for long-term gene delivery are inadequate. Controlled release systems using biocompatible and/or biodegradable polymers offer many advantages over conventional gene delivery approaches. We have characterized systems for controlled delivery of DNA from implantable polymer matrices (EVAc: poly (ethylene-co-vinyl acetate)) and injectable microspheres (PLGA and PLA: poly (D, L-lactide-co-glycolide) copolymer and poly (L-lactide), respectively). METHODS: Herring sperm DNA and bacteria phage lambda DNA were encapsulated as a model system. Released DNA concentration was determined by fluoroassays. Agarose electrophoresis was used to determine the dependence of release rate on DNA size. The Green Fluorescent Protein (GFP) gene was used to determine the integrity and functionality of released DNA. RESULTS: Both small and large DNA molecules (herring sperm DNA, 0.1-0.6 kb; GFP, 1.9 kb; lambda DNA, 48.5 kb) were successfully encapsulated and released from EVAc matrices, and PLGA or PLA microspheres. The release from DNA-EVAc systems was diffusion-controlled. When co encapsulated in the same matrix, the larger lambda DNA was released more slowly than herring sperm; the rate of release scaled with the DNA diffusion coefficient in water. The chemical and biological integrity of released DNA was not changed. CONCLUSIONS: These low cost, and adjustable, controlled DNA delivery systems, using FDA-approved biocompatible/biodegradable and implantable/injectable materials, could be useful for in vivo gene delivery, such as DNA vaccination and gene therapy. PMID- 10468037 TI - Determination of air content in protein microspheres. PMID- 10468036 TI - Pharmacokinetics and tissue disposition in monkeys of an antisense oligonucleotide inhibitor of Ha-ras encapsulated in stealth liposomes. AB - PURPOSE: This study examined the pharmacokinetics and tissue distribution of an antisense oligonucleotide ISIS 2503, formulated in stealth (pegylated) liposomes (encapsulated) or in phosphate-buffered saline (unencapsulated). METHODS: Encapsulated or unencapsulated ISIS 2503 was administered to rhesus monkeys by intravenous infusion. The concentrations of ISIS 2503 and metabolites in blood, plasma, and tissue samples were determined by capillary gel electrophoresis. RESULTS: Plasma concentrations of encapsulated ISIS 2503 decreased mono exponentially after infusion with a mean half-life of 57.8 hours. In contrast, the concentration of unencapsulated ISIS 2503 in plasma decreased rapidly with a mean half-life of 1.07 hours. Both encapsulated and unencapsulated ISIS 2503 distributed widely into tissues. Encapsulated ISIS 2503 distributed primarily to the reticulo-endothelial system and there were few metabolites observed. In contrast, unencapsulated ISIS 2503 distributed rapidly to tissue with highest concentration seen in kidney and liver. Nuclease-mediated metabolism was extensive for unencapsulated oligonucleotide in plasma and tissues. CONCLUSIONS: The data suggest that stealth liposomes protect ISIS 2503 from nucleases in blood and tissues, slow tissue uptake, and slow the rate of clearance from the systemic circulation. These attributes may make these formulations attractive for delivering oligonucleotides to sites with increased vasculature permeability such as tumors or sites of inflammation. PMID- 10468038 TI - New clinical observations in hemangiomas. AB - Much research in endothelial biology is aimed at developing methods to stimulate productive angiogenesis or inhibit unwanted angiogeneseis. Hemangiomas provide a model for endothelial proliferation and involution. This article is intended to update the reader with new information regarding hemangiomas of infancy, the most common tumor of childhood. Topics such as possible origin, management issues, and psychosocial stresses are addressed. This field is constantly changing, but an effort has been made to include most of the recently reported articles. Our hope is that this information will enable physicians caring for patients with hemangiomas to better address the concerns of their patients and families. PMID- 10468039 TI - Update on juvenile xanthogranuloma: unusual cutaneous and systemic variants. AB - Juvenile xanthogranuloma (JXG) is a well-recognized benign disorder of infancy and early childhood characterized by yellowish cutaneous nodules that spontaneously regress over months to years. In the vast majority of children, JXG is limited to the skin and requires no treatment. Over the past two decades, unusual cutaneous and systemic forms of JXG have been increasingly reported. JXGs have been discovered, usually unexpectedly, in every organ system of the body. Correct diagnosis is crucial to prevent unnecessary invasive diagnostic and therapeutic procedures. Unusual clinical and histological variants of JXG often require immunohistochemical studies and/or electron microscopy to establish the diagnosis. Nonlipidized, giant, intramuscular, subcutaneous, and clustered JXG are but some of the variants that are discussed in this article. The immunohistochemistry of JXG, current nosology, and hypotheses regarding the origins of JXG are also reviewed. PMID- 10468040 TI - Perioral dermatitis in children. AB - Perioral dermatitis is a unique skin disorder of childhood. Its exact origin is unknown; it is probably an idiosyncratic response to exogenous factors such as the use of a topical fluorinated corticosteroid or other substances on the face. It is uncommon but not rare. The age of affected children has ranged from 7 months to 13 years, with the median being in the prepubertal period. Boys and girls, blacks and whites are equally affected. Clinical features include the following: (1) absence of systemic symptoms; (2) periorificial distribution (perioral, perinasal, periorbital); (3) skin lesions that consist of flesh colored or erythematous inflammed papules, micronodules, and rare pustules; and (4) variable pruritus. Laboratory tests are negative. Histologically, it is indistinguishable from rosacea; there is a superficial perifollicular granuloma consisting of epitheliod cells, and lymphohistiocytic infiltrate, with occasional giant cells. The disease waxes and wanes for weeks and months. Treatment consists of discontinuing topical fluorinated corticosteroid use if any, and using topical metronidazole alone or in combination with either oral tetracycline or erythromycin depending on the child's age. A low-potency topical steroid may also be used to suppress the inflammation and to wean off the strong steroid. Perioral dermatitis in childhood is probably a juvenile form of rosacea. PMID- 10468041 TI - Morphea and localized scleroderma in children. AB - Localized scleroderma refers to a diverse spectrum of disorders that involve fibrosis of the skin. Children are more likely than adults to develop localized forms of scleroderma. This condition may have devastating effects on growth and development such as limb asymmetry, flexion contractures, and psychological disability. The pathogenesis of localized scleroderma is unknown but its possible relation to Borrelial infection is discussed. This article reviews associated laboratory and radiologic abnormalities, and discusses implications for monitoring disease activity. There is no universally effective therapy for this idiopathic condition and therapy is limited. A rationale for treatment based on disease subtype and severity is provided. PMID- 10468042 TI - Cutaneous T cell lymphoma in children. AB - Cutaneous T cell lymphoma (CTCL) is a term used to describe a heterogeneous group of lymphoproliferative disorders that involve the skin. Mycosis fungoides and Sezary syndrome are two forms of CTCL that arise in adults and are uncommon in children. Dermatologists frequently employ the term CTCL as a synonym for mycosis fungoides and Sezary syndrome. There has been increased recognition that the mycosis fungoides and Sezary syndrome forms of CTCL may arise in children and adolescents. Moreover, younger adults with these types of CTCL report onset of symptoms during adolescence. The literature regarding CTCL in childhood is limited. The purpose of this article is the review the literature regarding CTCL arising in children. The clinical presentation including morphologic variants, diagnosis, prognosis, and management are summarized. PMID- 10468043 TI - Laser treatment of pigmented lesions in children. AB - A variety of lasers can be used to effectively treat cutaneous pigmentation in children. Furthermore, using the principle of selective photothermolysis, several pigment-specific lasers can be used to achieve significant clinical improvement if not complete removal of pigmented lesions with greatly reduced risks of scarring, textural or pigmentary changes. A summary of laser treatments for pigmented lesions that commonly present in children including cafe-au-lait macules, nevus of Ota and other dermal melanocytosis, congenital melanocytic nevus, nevus spilus, lentigines, epidermal nevus, Becker's nevus, and tattoos is presented. Overall, laser treatment is a very safe, effective treatment option that could offer distinct advantages compared with other conventional modalities. PMID- 10468044 TI - Cosmetics, skin care, and appearance in teenagers. AB - Adolescence is a period of tremendous transformation in the appearance of the body and the evolution of the mind that will eventually lead to adulthood. "Yesterday's child" will need to assume and exert control over these changes. Therefore his/her appearance becomes a means of communication, a language to express his/her pursuit of self-identity. The cosmetic industry has identified teenagers as "powerful" consumers, and offers them various toiletry and skin care products that should fulfill their needs, such as cleansing, hydrating, and photoprotective agents. Certain decorative cosmetics, especially for hair and nails, are attractive to them also. For some teenagers, the expression of individualism is through body art such as tattooing and body piercing. Areas of concern are the lack of motivation for sun protection and the risky behavior associated with body piercing and tattooing. PMID- 10468045 TI - Novel roles of nitric oxide in hemorrhagic shock. AB - The aim of the current article is to overview the recent developments in the field of hemorrhagic shock research, as it relates to the roles of nitric oxide (NO) in the pathogenesis of this condition. The first part of the review focuses on the roles of peroxynitrite, a reactive oxidant produced from the reaction of NO and superoxide. The second part of the review deals with the novel findings related to the recently identified regulatory roles of the inducible isoform of nitric oxide synthase (iNOS) in the expression of pro-inflammatory mediators in hemorrhagic shock. (1) The role of peroxynitrite: Immunohistochemical and biochemical evidence demonstrate the production of peroxynitrite in hemorrhagic shock. Peroxynitrite can initiate a wide range of toxic oxidative reactions. These include initiation of tyrosine nitration, lipid peroxidation, direct inhibition of mitochondrial respiratory chain enzymes, inactivation of glyceraldehyde-3-phosphate dehydrogenase, inhibition of membrane sodium/potassium ATP-ase activity, inactivation of membrane sodium channels, and other oxidative modifications of proteins. All these toxicities are likely to play a role in the pathophysiology of hemorrhagic shock. A combined anti-inflammatory agent, mercaptoethylguanidine, which selectively inhibits iNOS and scavenges peroxynitrite, prevents the delayed vascular decompensation and the cellular energetic failure associated with late hemorrhagic shock. Peroxynitrite is a potent trigger of DNA single strand breakage, with subsequent activation of the nuclear enzyme poly (ADP ribose) synthetase (PARS), leading to eventual severe energy depletion of the cells, and necrotic-type cell death. Pharmacological inhibition of PARS, with 3-aminobenzamide or 5-iodo-6-amino-1,2-benzopyrone, improves hemodynamic status and prolongs survival time in rodent and porcine models of severe hemorrhagic shock. (2) Novel signaling roles of induced NO in hemorrhagic shock. Although the severity and duration of shock may dictate the timing and extent of iNOS expression, it is now evident that the up-regulation of iNOS can take place during sustained shock. Accumulated data indicate that iNOS expressed during shock contributes to vascular decompensation, as classically described by Wiggers. In addition, the presence of even low levels of iNOS at the time of resuscitation enhances the inflammatory response that follows the reperfusion state. Pharmacological inhibition of iNOS with N6-(iminoethyl)-L lysine or genetic inactivation of iNOS (iNOS knockout mice) attenuates the activation of the transcription factors nuclear factor kappa B (NFkappaB) and Signal Transducer and Activator of Transcription 3 (STAT3), and ameliorates the increases in interleukin-6 and G-CSF messenger RNA levels in the lungs and liver. Inhibition of iNOS results in a marked reduction of lung and liver injury produced by hemorrhagic shock. Thus, induced nitric oxide, in addition to being a "final common mediator" of hemorrhagic shock, is essential for the up-regulation of the inflammatory response in resuscitated hemorrhagic shock. Furthermore, a picture of a pathway is evolving that contributes to tissue damage both directly via the formation of peroxynitrite, with its associated toxicities, and indirectly through the amplification of the inflammatory response. PMID- 10468046 TI - Transient monocyte release of interleukin-10 in response to traumatic brain injury. AB - A significant component of the immune response to trauma results in the systemic presence of cytokines which have the potential to suppress the patient's immune response to infection and contribute to post-injury complications. We assayed peripheral blood leukocytes obtained from 10 patients with head trauma to determine their production of interleukin (IL). Serum was assayed for the presence of IL-10, TGFbeta1, and IFNgamma by ELISA. Peripheral blood leukocytes were screened for intracellular IL-10 and IFNgamma by fluorescence-activated flow cytometry, and cytokine-specific mRNA was detected by the polymerase chain reaction. We detected an immediate, but transient, presence of IL-10 in the sera of all 10 patients who suffered head trauma. IL-10-specific intracytoplasmic immunofluorescence was also detected immediately after injury in peripheral blood monocytes, but not in lymphocytes or granulocytes. IL-10-specific mRNA was detected in peripheral blood leukocytes in only 50% of patients immediately after injury, when the highest serum levels of IL-10 were observed. Our data indicates that release of pre-formed IL-10 by monocytes contributes to the presence of IL 10 found in patient peripheral blood immediately after head injury. PMID- 10468047 TI - Influence of granulocyte-macrophage colony-stimulating factor (GM-CSF) on whole blood endotoxin responsiveness following trauma, cardiopulmonary bypass, and severe sepsis. AB - Major surgery, multiple injury, and severe sepsis lead to an impaired immune response. The suppressed status of the immune system is reflected by a reduced TNFalpha production of whole blood after stimulation with endotoxin in vitro and by a decreased HLA-DR expression on monocytes. In the present study, the effect of the immunostimulating hematopoetic growth factor GM-CSF on whole blood cultures of multiple injury, cardiac surgery, and severe sepsis patients was investigated. Endotoxin-induced TNFalpha production and HLA-DR expression was reduced in blood cultures of these patients compared to healthy donors. Preincubation with GM-CSF in vitro increased cytokine production in volunteers' and all patients' blood specimens in a dose-dependent manner. The elevation of cytokine response in cardiopulmonary bypass patients' blood, caused by in vitro preincubation with GM-CSF, equaled that of normal patients, whereas GM-CSF caused a lower rise of TNFalpha-producing capacity in blood of multiple-injury and sepsis patients. Further, GM-CSF treatment in vitro increased the down-regulated HLA-DR expression on monocytes prepared after cardiac surgery to a degree comparable to preoperative levels. Finally, GM-CSF incubation in vitro elevated TNFalpha synthesis in normal monocytes and in cells treated with a combination of the anti-inflammatory mediators IL-10, TGFbeta, and PGE2. These experiments show that hyporesponsiveness of whole blood induced by trauma, sepsis, or cardiac surgery is not irreversible but can be, at least in vitro, overridden by the immunostimulating compound GM-CSF. PMID- 10468048 TI - Reduction of infarct size in the rat heart by LPS preconditioning is associated with expression of angiogenic growth factors and increased capillary density. AB - Inflammation induces the expression of angiogenic growth factors in tissues, which leads to microvascular growth. Bacterial lipopolysaccharide (LPS) provokes a transient inflammatory response in the heart and induces delayed cardiac resistance to post-ischemic contractile dysfunction. In this study, we examined: 1) the effects of LPS on myocardial expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), 2) whether an increase in the density of myocardial microvessels follows the expression of angiogenic growth factors, and 3) the effect of LPS on myocardial resistance to infarction and its relationship with microvascular growth. Rats were treated with LPS (from Salmonella typhimurium, 0.5 mg/kg i.p.). The expression of bFGF and VEGF in the myocardium was examined at 6 and 12 h after LPS treatment by immunofluorescent staining. Myocardial capillary and arteriole densities were determined 3 days after LPS treatment by morphometry, using immunofluorescent staining of von Willebrand factor (a marker protein of endothelial cells) and alpha-smooth muscle actin (a marker protein of smooth muscle cells). To examine cardiac resistance to infarction, hearts were subjected to 40 min of regional ischemia and 2 h of reperfusion by reversible occlusion of left coronary artery at 3 days after LPS treatment. LPS induced cardiac bFGF and VEGF at 6 and 12 h after treatment. The expression of these growth factors was followed by an increase in myocardial capillary density (2032 +/- 78/mm2 vs. 1617 +/- 47/mm2 in saline control, P < 0.05), but not arteriole density, at 3 days. Meanwhile, infarct size was significantly reduced by LPS preconditioning (infarct/left ventricle 12.3 +/- 1.04% vs. 21.7 +/- 1.65% in saline control, 43% reduction, P < 0.05). These results suggest that LPS preconditioning induces cardiac bFGF and VEGF, and an increase in myocardial capillary density. This increased myocardial capillary density is associated with a reduced infarct size after in vivo regional ischemia reperfusion. PMID- 10468049 TI - Neutrophil heat shock protein expression and activation correlate with increased apoptosis following transmigration through the endothelial barrier. AB - Polymorphonuclear leukocytes (PMN) undergo endothelial transmigration upon activation or in response to a chemoattractant. Such cells are stressed and have an increased capacity to incite tissue injury. Little is known about the effect of transmigration on PMN stress gene responses, PMN activation, and ultimately programmed cell death (apoptosis). Human endothelial cells (ECV-304) were plated onto transwell membranes to form an endothelial monolayer and PMN transendothelial migration through this endothelial barrier was examined. Chemotaxis was induced by formyl-methionyl-leucyl-phenylalanine (fMLP). Flow cytometry was used to determine PMN receptor expression (CD11b, CD14, CD16, CD18, CD54), phagocytosis, and apoptosis. Heat shock protein (Hsp) expression was evaluated by Western blotting. fMLP-induced PMN transendothelial migration resulted in increased adhesion receptor expression and phagocytosis. Migrated PMN also had an increased rate of apoptosis as evaluated by uptake of propidium iodide and decreased FcgammaR III (CD16) expression. Increased PMN apoptosis coincided with induction of Hsp72 following transmigration. Thus, naive PMN that migrate through endothelium in response to a chemoattractant undergo activation as represented by increased phagocytosis and expression of adhesion receptors. PMID- 10468050 TI - Severe injury triggers antigen-specific T-helper cell dysfunction. AB - Although it is established that post-injury immune dysfunction involves alterations in T-cell function, the effects of injury on T-cell function in vivo are poorly understood. This study uses a mouse injury model and an antigen immunization approach to investigate the influence of injury on antigen-specific T-helper cell function. We report here that injury triggered a significant reduction in antigen-specific T-helper-1 (Th1)-dependent IgG2a antibody formation, while IgM, IgG1, and IgE production was unchanged. In addition, injury caused a reduction in cytokine production (IL-2, IFNgamma and IL-10) by antigen stimulated T-cells. We also demonstrate that interleukin 12 (IL-12), a cytokine that promotes Th1 cell differentiation, restored IgG2a antibody formation and corrected the injury-induced reduction in antigen-stimulated cytokine production. Taken together, these findings indicate that severe injury induces a dramatic reduction in Th1 cell function in vivo and suggest that therapies designed to restore Th1 cell function may be beneficial to the injured host. PMID- 10468051 TI - An isoenzyme-selective inhibitor of phosphodiesterase 4 and 1, KF19514, may be useful in the treatment of systemic anaphylaxis: an in vivo study in rabbits. AB - The present study was conducted to determine whether an inhibitor of phosphodiesterases 4 and 1, KF19514, is useful in the treatment of systemic anaphylaxis in fentanyl-anesthetized rabbits. Eighty-two rabbits were randomly allocated to 7 groups. Groups I-1 (0.01 mg x kg(-1)), I-2 (0.1 mg x kg(-1)), and I-3 (1 mg x kg(-1)) received KF19514 10 minutes before antigen challenge, with Group II serving as control. Group IV and Group V received KF19514 and aminophylline 1 minutes after antigen challenge, respectively, with Group III serving as control. The survival rate was higher in Groups I than in Group II, rates were similar in Groups I-1, I-2, and I-3. The survival rate was also higher in Group IV than in Group III. Pulmonary resistance (R(L)) was significantly lower in Groups I-2 and I-3 than in Group II. Dynamic pulmonary compliance (C(dyn)) was significantly higher in Group I-3 than in Group II. Heart rate and central venous pressure were similar in Groups I and II. In Groups III, IV, and V, heart rate and mean arterial pressure were comparable, but central venous pressure in Group IV was significantly lower than in Group V. In conclusion, the administration of KF19514, an inhibitor of PDEs 4 and 1, to rabbits either before or after antigen challenge improved bronchoconstriction provoked by systemic anaphylaxis with minimal concomitant cardiovascular side effects compared with aminophylline, suggesting that this agent may be useful in the treatment of systemic anaphylaxis. PMID- 10468052 TI - Glycine improves survival after hemorrhagic shock in the rat. AB - This study investigated the effect of glycine on hemorrhagic shock in the rat. Rats were bled to maintain mean arterial pressure at 30-35 mm Hg for 1 h and subsequently resuscitated with 60% shed blood and lactated Ringer's solution. Only 20% of rats receiving saline just prior to resuscitation survived 72 h after shock. Survival was increased by glycine (11.2-90.0 mg/kg, i.v.) in a dose dependent manner (half-maximal effect = 25 mg/kg) and reached maximal values of 78% at 45 mg/kg. Eighteen hours after resuscitation, creatinine phosphokinase increased 23-fold, transaminases increased 33-fold, and creatinine was elevated 2.4-fold, indicating injury to the heart, liver, and kidney, respectively. Pulmonary edema, leukocyte infiltration, and hemorrhage were also observed. In the kidney, proximal tubular necrosis, leukocyte infiltration, and severe hemorrhage in the outer medullary area occurred in rats receiving saline. Glycine reduced these pathological alterations significantly. It has been reported that oxidative stress and tumor necrosis factor(TNF)-alpha-production are involved in the pathophysiology of multiple-organ injury after shock. In this study, free radical production was increased 4-fold during shock, an effect blocked largely by glycine. Increases in intracellular calcium and production of TNF-alpha by isolated Kupffer cells stimulated by endotoxin were elevated significantly by hemorrhagic shock, alterations which were totally prevented by glycine. Taken together, it is concluded that glycine reduces organ injury and mortality caused by hemorrhagic shock by preventing free radical production and TNF-alpha formation. PMID- 10468053 TI - Evidence of multi-step regulation of HSP72 expression in experimental sepsis. AB - Heat shock proteins (Hsps) are a family of highly conserved proteins induced in response to various stresses. Hsps protect cells against subsequent lethal circumstances. Previous work from our laboratory has indicated that Hsp72 is not induced during experimental sepsis in rats, but the regulation of the induction of Hsp72 synthesis in this disease cascade has not been investigated. In the present study, we evaluated the expression of the hsp72gene, focusing on the activation and DNA-binding ability of heat shock factor 1 (HSF1), hsp mRNA accumulation, and Hsp72 synthesis in animal sepsis models induced by cecal ligation and puncture procedure. The results were compared with those of sham treated and heat-shocked rats. It was shown that the expression of the hsp72 gene in sepsis was a multi-step process, as previously documented in in vitro studies. Hsp72 synthesis was not induced during sepsis, whereas DNA binding of HSF was detectable, suggesting that the induction of Hsp72 is blocked downstream to HSF DNA complex formation by the metabolic alteration occurring during sepsis. The dissociation failure of the constitutive heat shock element binding factor (CHBF) from the heat shock element may play an important role in this negative regulation. PMID- 10468054 TI - Effects of N(omega)-nitro-L-arginine (L-NOARG) on blood flow and vasomotion in rat diaphragm microcirculation during hemorrhagic hypotension. AB - The role of N(omega)-nitro-L-arginine (L-NOARG), a nitric oxide (NO) synthase inhibitor, in the control of blood flow and vasomotion in rat diaphragm microcirculation during hemorrhagic hypotension was investigated by means of laser Doppler flowmetry (LDF). Fifty-six Sprague-Dawley rats were divided into seven groups. Ten minutes after one-stage hemorrhage to 40-60% of initial blood pressure, the rats received 15 min topical superfusion of saline (group 1, time control), 0.1 mM L-NOARG (group 2), 10 mM L-arginine (group 3), or vehicle (0.1% DMSO and 0.9 mN NaOH, group 4). For groups 5 and 6, L-NOARG or its vehicle was superfused for 15 min without hemorrhage. In group 7, the vasodilator responses to the endothelium-dependent vasorelaxant acetylcholine (ACH) and the endothelium independent vasorelaxant sodium nitroprusside (SNP) were assessed at rest and after 25 min of hemorrhagic hypotension. The results showed no significant differences in blood flow, fundamental frequency, or relative amplitude of the rat diaphragm microcirculation before or after administration of the test agents among the first four groups during hemorrhagic hypotension or in groups 5 and 6 during sham operation without hypoperfusion. Hemorrhagic hypotension significantly decreased the vasodilator response to ACH (p = 0.003), but not to SNP. We conclude that NO did not play an important role in the regulation of blood flow or vasomotion in rat diaphragm microcirculation during acute hemorrhagic hypotension. PMID- 10468055 TI - Vascular permeability increase and plasma volume loss induced by endotoxin was attenuated by hypertonic saline with or without dextran. AB - Endotoxin given intravenously is known to cause plasma leakage and subsequent loss of circulating plasma volume. Hypertonic saline resuscitation has been successfully applied in hemorrhagic and traumatic shock, but its application for the treatment or prevention of septic or endotoxin shock is less well studied. Our aim was to investigate the effects of endotoxin on plasma leakage in hamsters when administered in two different ways: applied locally to the hamster cheek pouch microcirculation or systemically by i.v. injection. The cheek pouch was studied by intravital microscopy using FITC-labeled dextran as a tracer of plasma leakage. Escherichia coli lipopolysaccharide (LPS) was continuously added into the superfusion buffer of the cheek pouch preparation during 120 min in two control groups (each n = 6) and two further groups (each n = 6) treated with either hypertonic saline (HS) or hypertonic saline and dextran (HSD). Treatment was given as an i.v. injection 0.35 mL NaCl 7.5%/100 g b.w. during 4 min starting 15 min prior to the start of endotoxin application. Endotoxin caused a reversible increase in the number of postcapillary venular leaks with a maximal response at 70 min after start of endotoxin application. The maximal responses were reduced to 36% in the HS-treated and to 37% in the HSD-treated group in comparison to what was seen in the control groups. In the second part of the study endotoxin was given i.v. 0.3 mg/kg to anesthetized hamsters (n = 41) and arterial blood samples were collected at 0, 60, 120, and 180 min after endotoxin injection for measurement of hematocrit and plasma FITC-dextran concentration. Hamsters were divided into seven groups: untreated control group (n = 6); HSC control group given only an i.v. injection of hypertonic saline (n = 6); LPS group given endotoxin 0.3 mg/kg during 1 min (n = 9); HSp group given hypertonic saline (NaCl 7.5%) 10 min prior to i.v. endotoxin (n = 6); HSa group given hypertonic saline 10 min after i.v. endotoxin (n = 6); HSD group given hypertonic saline with dextran 40, 10 min prior to i.v. endotoxin (n = 6); HSD control group given only i.v. hypertonic saline + dextran and no endotoxin (n = 2). Injection of endotoxin caused a significant increase in hematocrit, which was counteracted by hypertonic saline treatment, with or without dextran, probably due to reduced extravasation of plasma in postcapillary venules. PMID- 10468056 TI - Incarceration of the retroverted gravid uterus--a review. PMID- 10468057 TI - Antioxidant role of endogenous coenzyme Q against the ischemia and reperfusion induced lipid peroxidation in fetal rat brain. AB - BACKGROUND: Ischemia and subsequent reperfusion induce lipid peroxidation in the cerebrum of the fetal rat. The present study evaluated the antioxidant activity of endogenous coenzyme Q in protecting against the lipid peroxidation induced in the fetal rat brain by ischemia/reperfusion. METHODS: We used wistar rats at day 19 of pregnancy. Fetal ischemia was induced by bilateral occlusion of the utero ovarian artery for 20 minutes. For reperfusion, the occlusion was released and the circulation was restored for 30 minutes. Control rats underwent sham operation. We determined the levels of thiobarbituric acid-reactive substances, the concentrations of coenzyme Q9, coenzyme Q10, and the mitochondrial respiratory control index in fetal brains. RESULTS: Occlusion for 20 minutes significantly reduced the respiratory control index (p < 0.01), but did not alter the levels of thiobarbituric acid-reactive substances, coenzyme Q9 or coenzyme Q10. Subsequent reperfusion, however, significantly increased the level of thiobarbituric acid-reactive substances (from 6.53+/-1.54 to 11.46+/-3.31 nM/mg of protein, p < 0.01) and significantly decreased the level of coenzyme Q9 (from 291.73+/-108.94 to 162.44+/-56.83 pM/mg of protein, p < 0.05) and that of coenzyme Q10 (from 153.10+/-75.24 to 79.84+/-30.40 pM/mg of protein, p < 0.05). The respiratory control index was still significantly lower following reperfusion than in controls (p < 0.01). Significant negative correlations were observed between the level of thiobarbituric acid-reactive substances and the concentrations of either coenzyme Q9 (r = -0.68, p < 0.001) or coenzyme Q10 (r = 0.70, p < 0.001). CONCLUSION: Endogenous coenzyme Q may protect the fetal rat brain against the lipid peroxidation induced by ischemia/reperfusion. PMID- 10468058 TI - Differential levels of T helper cytokines in preeclampsia: pregnancy, labor and puerperium. AB - BACKGROUND: There are suggestions that T helper 1 cytokines may be detrimental to early pregnancy and T helper 2 cytokines protective of the pregnancy. Their role in preeclamptic pregnancy, labor and puerperium, is not clear. MATERIALS AND METHODS: Twenty-eight preeclamptic women and their matched controls were evaluated, at the Departments of Obstetrics and Gynecology, Faculty of Medicine, Kuwait University and Maternity Hospital, Kuwait. Outcome measures evaluated were serum levels of TNF alpha and interleukin-4 at 12, 24, 36 weeks of gestation, advanced labor and at 1 hour and daily postpartum until they were undetectable, using ELISA technique. RESULTS: T helper cytokines showed higher serum levels in preeclampsia than normotensive pregnancy (p < 0.01, 0.01), in established labor (p < 0.05) and at 1 hour postpartum (p < 0.01 for IL-4) and p < 0.02 for TNF alpha. There was significant increase of IL-4 between 12 to 24 weeks in normal pregnancy compared to preeclampsia (p < 0.001) but not for TNF alpha. By 24 hours postpartum, IL-4 was still detectable in eight parturients compared to one patient with detectable TNF alpha (p < 0.04). Detectable IL-4 levels after 24 hours postpartum were associated with intrauterine growth retardation (p < 0.03). CONCLUSION: IL-4 has a dichotomous role in pregnancy. Normotensive pregnancy is associated with high increase in IL-4 in the first half of the pregnancy, but in the second half of pregnancy and puerperium, high levels of IL-4 are associated with preeclampsia. PMID- 10468059 TI - Pregnancy-related back and pelvic pain and changes in bone density. AB - OBJECTIVE: To elucidate whether there is an association between pregnancy-related back and pelvic pain and changes in bone density. METHODS: In this prospective cohort study, bone density was measured in the distal and ultra-distal forearm at 12 and 35 weeks of pregnancy and at 5 months post partum. The location and degree of any back or pelvic pain was registered. The patients were classified into four subgroups on the basis of presence or absence of disabling pain in late pregnancy and presence or absence of pain at 5 months' follow-up. Forty-nine women participated. RESULTS: Bone density decreases during pregnancy and lactation. Trabecular bone is mainly lost during pregnancy and cortical bone during lactation. No association between back or pelvic pain during pregnancy and bone loss was found. Between 35 weeks of pregnancy and 5 months post partum, bone loss in all the women was estimated as 1.1% of cortical bone (p < 0.001) and 0.6% of trabecular bone (n.s.). During the same period five women with mild pain during pregnancy and pain at follow-up lost 3.9% of cortical bone (p=0.043) and 5.3% of trabecular bone (p=0.043). Although this bone loss was significant compared to the other subgroups, the small study size does not permit general conclusions to be drawn from this finding. CONCLUSION: The results indicate that bone density decreases during pregnancy and lactation. The decrease in bone density was not associated with back or pelvic pain during pregnancy. It remains unclear whether bone loss is associated with back and pelvic pain during lactation. PMID- 10468060 TI - Prescribing during pregnancy and lactation with reference to the Swedish classification system. A population-based study among Danish women. The Euromap Group. AB - AIM: To assess the current prescribing pattern for 15,756 primiparae before, during, and after their pregnancies with reference to fetal and neonatal risk. METHOD: A prescription database study with linkage to The Danish Medical Birth Registry from 1991 to 1996. The drug subsidy system in Danish retail pharmacies, made it possible to identify prescriptions by individual use. All 34,334 prescriptions were set against the Swedish classification of risk of drug use in pregnancy and lactation. RESULTS: During pregnancy, safe (group A), potentially harmful (group B3, C, and D), and non-classifiable drugs accounted for 40.9%, 26.6% and 28.7% respectively. The proportion of women who redeemed drugs was 29.2%, 8.6%, 18.7% and 0.9% from drug groups A, B, C and D respectively. The proportion of prescriptions from high risk groups declined during the course of pregnancy. Postpartum, safe drugs (group I and II), drugs with possible harmful neonatal effects (group III), and non-classifiable drugs accounted for 43.5%, 4.8%, and 35.8% of the prescriptions, respectively. CONCLUSION: According to the Swedish classification system, we found that during pregnancy and lactation a high proportion of Danish women were exposed to one or more drugs in high risk groups; furthermore, knowledge regarding their safety for the fetus and neonate was limited for a large proportion of the prescriptions. Current evidence about long-term effects of prenatal exposure stresses the need for long-term follow-up of health and development among exposed children. PMID- 10468061 TI - Moderate smoking during pregnancy is associated with a reduced risk of preeclampsia. AB - BACKGROUND: To investigate the effects of maternal smoking during pregnancy on the development of preeclampsia (PE). METHODS: The study comprised two populations with singleton pregnancies, one collected from 1990 to 1994 at the Malmo University Hospital (Malmo series, n=14,510) and the other from the National Birth Registry of Sweden 1993 (National series, n=113,211). Women with PE (n=281 and n=2,865, respectively) were compared to those without PE, delivered in 1993 (n=2,811 and n=110,346, respectively). The subgroups of women who had PE associated with preterm birth (n=58 and n=693, respectively) were compared with the same control groups in both series. RESULTS: Multiple logistic regression analysis showed that, in comparison with non-smokers, moderate smokers (1-9 cigarettes per day) were characterized by a lower incidence of PE (odds ratio (OR) 0.4; 95% confidence interval (CI) 0.22-0.60), and PE associated with preterm birth (OR 0.1; 95% CI 0.01-0.67). The corresponding figures in the validating National series were also significantly lower for moderate smokers (OR 0.6; 95% CI 0.5-0.7 and OR 0.6; 95% CI 0.5-0.8, respectively). CONCLUSION: Moderate smoking during pregnancy seems to protect against the development of PE and PE associated with preterm birth. Nicotine might be the agent responsible for this protective effect. PMID- 10468062 TI - Selective arterial embolization of the uterine arteries in the management of intractable post-partum hemorrhage. AB - BACKGROUND: To evaluate the efficacy and safety of selective arterial embolization in the management of intractable post-partum hemorrhage. METHODS: Thirty-five consecutive women with severe post-partum hemorrhage (primary, n=25; secondary, n=10) were treated by selective embolization of the uterine arteries. The main cause of immediate post-partum hemorrhage was atonic uterus. Retained placental fragments with endometritis was the main cause of delayed hemorrhage. In all cases, hemostatic embolization was performed because of intractable hemorrhage. Hysterectomy had been performed in two cases before embolization but it had also failed to stop the bleeding. RESULTS: Angiography revealed extravasation in ten cases, spasm of the internal iliac artery in four cases, false aneurysm in two cases and arteriovenous fistula in one case. After embolization, immediate cessation or dramatic diminution of bleeding was observed in all cases. Two patients required repeated embolization the following day. No major complication related to embolization was found. In one patient with placenta accreta, delayed hysterectomy was necessary. Normal menstruation resumed in all women but two who had hysterectomy. One woman became pregnant after embolization. CONCLUSION: Selective emergency arterial embolization is an effective means of controlling severe post-partum hemorrhage. This procedure avoids high risk surgery and maintains reproductive ability. PMID- 10468063 TI - Women's anticipations of and experiences with induction of labor. AB - BACKGROUND: Although induction of labor is a common procedure, there are limited data on its psychoemotional effects. We studied women's expectations of and experiences with labor induction in a large university hospital. PATIENTS AND METHODS: A total of 296 parturients were recruited, with 270 (91%) taken into the final analysis. Of these, 135 underwent elective induction of labor (44 with amniotomy and 91 with cervical ripening by the use of vaginal prostaglandin gel, followed by amniotomy), whereas the remaining 135 women gave birth spontaneously and served as controls. Each woman was interviewed as to her knowledge, expectations, and feelings about labor before induction or at start of labor (controls) as well as after delivery with the help of questionnaires containing yes/no or multiple-choice questions and visual analog scales. RESULTS: The induction and control groups, in regard to baseline clinical characteristics, did not differ. Labor ended in cesarean section for 24 women in the induction group (18%) and for 12 women in the control group (9%) (p=0.04); these women were included in the data analysis. In the induction group, 67%, and in the control group, 48% of women (p=0.002) reported having received sufficient information on labor induction from medical personnel at prenatal or obstetric clinics. Attitudes towards induction of labor were antenatally positive in 78% of women in the induction group and in 69% in the control group. The induction group reported fear of pain less often (45%) than the control group (57%) (p=0.03). In the induction group, 76 women (56%), and in the control group 94 women (70%) (p=0.02), wanted to participate in the decision on the method of induction, and 74% and 83%, respectively, wished to influence the timing of induction. When interviewed post partum, the labor experience corresponded with the patients' expectations better in the control than in the induction group (p=0.03). Labor induction was a positive experience in 90% of women who underwent immediate amniotomy and in 69% of those who received prostaglandin ripening first. CONCLUSION: Labor induction was a positive experience only seldom eliciting negative feelings. A third of the parturients undergoing induction need more information on the procedure and most want to participate in decision-making concerning the method and timing of induction. PMID- 10468064 TI - Delayed interval delivery in multiple pregnancies. Is optimism justified? AB - OBJECTIVE: Evaluation of the success of delayed interval delivery in multiple pregnancies. METHOD: Retrospective analyses of all multiple pregnancies with the birth of one child at a gestational age of 16-31 weeks (n=80). Three groups were defined: group I, unsuccessful attempt to temporize delivery, group II, interval between the first child and the other child or children 3 hours or more, with delivery of the remaining children before 28 weeks and group III, prolongation of pregnancy beyond 28 weeks. RESULTS: In 15 multiple pregnancies (ten sets of twins, five sets of triplets), the aim was to postpone the delivery of the second (and third) child. The mean gestational age at admittance was 25 weeks. Delay of the delivery of the second child was achieved in ten cases, five in group II and five in group III. In groups II and III, the mean interval of delivery was 12 days; the gestational age at the delivery of the remaining children in these ten patients was 27 5/7 weeks. No relation could be found between the reason for admittance and the interval in delivery between the first and the other children. The only severe maternal complication was blood loss exceeding 2 liters. In the neonatal outcome of first and second children (with a gestational age of 28 weeks or more) no striking differences were observed. CONCLUSION: Delayed delivery in multiple pregnancies after the birth of the first child is only successful in a minority of patients and should be considered prior to the birth of the first child. PMID- 10468065 TI - Health effects of long-term use of the intrauterine levonorgestrel-releasing system. A follow-up study over 12 years of continuous use. AB - BACKGROUND: This report comprises women from one center, who all participated in a larger European multi-center study, which included 2758 healthy women, of whom 1821 were randomly allocated to the levonorgestrel-releasing intrauterine system (Lng-IUS) and 937 to a copper-IUD (Nova-T). At the end of the 5 year study period, 109 of the 300 women, who originally were allocated to the Lng-IUS at this center, were still using the Lng-IUS. The women were offered the opportunity to continue the Lng-IUS and 100 women chose to do so. After a total period of almost 7 years of continuous use, a second Lng-IUS was offered and 82 of the women chose to start a second segment of Lng-IUS use. After another 5 years, 69 of the women were fitted with their third Lng-IUS. METHODS: These 82 women were examined yearly throughout the study period and bleeding pattern, hemoglobin, weight, blood-pressure, general well-being and intercurrent disease were analyzed to assess long-term effects. RESULTS: No pregnancy occurred during the follow-up. One case of partial expulsion occurred and one case of pelvic inflammatory disease. Seventy-seven percent of the women did not report any health problems at all during the entire follow-up period of 13 years. At the end of the second period with the Lng-IUS, 60% reported amenorrhea, 12% infrequent, scanty bleeds and 28% regular, scanty bleeds. As a consequence, the hemoglobin values increased, with a mean increase, after 12-13 years, being 1.35 g/dl. Blood pressure increased slightly and body weight increased with a mean increase of 0.49 kg/year. Seven women became postmenopausal during the follow-up and started estrogen replacement therapy with the Lng-IUS in situ. CONCLUSIONS: This follow up study suggests that the Lng-IUS remains a safe and effective method of contraception, allowing women prolonged relief of menstrual problems, and for women in their late reproductive years, offering a convenient and bleeding-free transition into the menopause. PMID- 10468066 TI - The bacteriological changes after re-using a vaginal continence guard. AB - BACKGROUND: Involuntary loss of urine is a major health care problem, especially in women. The Conveen Continence Guard is a non-surgical treatment of stress incontinence. A possible risk of vaginal devices is introduction of changes in bacteriological flora of the vagina, especially after re-use of the guard. Moreover, vaginal devices can induce magnesium ion deficiency and thereby facilitate growth of Staphylococcus Aureus. This can lead to the production of toxin-1, eventually resulting in toxic shock syndrome. METHODS: The first aim of this study was to investigate the capacity to absorp magnesium ions in an in vitro study. Secondly, 14 patients with stress incontinence were examined for bacteriological changes in the vagina after re-using the continence guard for four days. After completing this study the patients were asked whether re-using the continence guard was as convenient as single use. RESULTS: The continence guard had no absorption capacity for magnesium ions. The vaginal bacteriological changes after re-use showed in 11% changes in bacterial growth and in 20% increased bacterial growth. Only one patient developed a vaginal colonization with Staphylococcus Aureus but had no symptoms of disease. The continence guard showed itself to be less effective in preventing stress incontinence after re use. CONCLUSIONS: Although re-use of the continence guard is feasable from a bacteriological point of view, re-use renders the device less effective. PMID- 10468067 TI - Co-expression of vascular endothelial growth factor and thymidine phosphorylase in endometrial cancer. AB - OBJECTIVE: To examine expression of vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP) together with microvessel count in endometrial cancer, and to investigate the relationship with clinicopathological and biological factors. METHODS: VEGF expression, TP expression, the microvessel count (factor VIII-related antigen positive cells), bcl-2 expression, proliferating cell nuclear antigen (PCNA) index, and p53 expression were determined in 50 resected endometrial cancer specimens, using immunohistochemistry. The relationship between VEGF and TP expression and correlation between expression and microvessel count were also given attention. These 3 factors were analyzed with regard to clinicopathological factors, bcl-2 expression, PCNA index, and p53 expression. RESULTS: Staining status of VEGF and TP was identical in 37 (74%) of 50 tumors, the correlation being statistically significant (p < 0.01). Combined analysis of VEGF and TP status showed that tumors which were VEGF-positive and/or TP-positive had a significantly higher microvessel count than did tumors which were both VEGF-negative and TP-negative (p < 0.001 and p < 0.05, respectively). TP expression correlated with bcl-2 expression, and VEGF expression inversely correlated with the PCNA index. Although clinical stage (p < 0.01), PCNA index (p < 0.01), and p53 expression (p < 0.01) significantly correlated with disease-free survival, neither VEGF/TP expression nor microvessel count contributed to prognostic estimates. CONCLUSIONS: VEGF and TP may cooperatively promote angiogenesis in endometrial cancer, but these expressions may have limited additional prognostic value. PMID- 10468068 TI - Labor abnormalities as a risk factor for shoulder dystocia. PMID- 10468069 TI - Recurrent incarceration of the retroverted gravid uterus--a case report. PMID- 10468070 TI - Successful management of malignant hyperthermia susceptibility during cesarean hysterectomy for postpartum hemorrhage. PMID- 10468071 TI - Anti-shock trousers (MAST) and transcatheter embolization in the management of massive obstetrics hemorrhage. A report of two cases. PMID- 10468072 TI - Four-year survival of patients with acute coronary syndromes without ST-segment elevation and prognostic significance of 0.5-mm ST-segment depression. AB - We prospectively evaluated all patients admitted to our coronary care unit during 1993 with ischemic chest pain but without ST-segment elevation on the presenting electrocardiogram, and determined the influence of the extent of ST-segment depression, measured using calipers and blinded to the outcome, on 4-year survival. The presenting symptoms of 367 patients (mean age 64 years) were coded according to the Braunwald classification, 86% being in class IIIB (primary unstable angina with rest angina within 48 hours) and 7.4% in class IIIC (postinfarction angina). Thirty-two patients (8.6%) had myocardial infarction at presentation (defined as a creatine kinase level exceeding twice the reference range within 18 hours). During hospitalization 97% of patients received aspirin, 67% received intravenous heparin, 37% underwent angiography, and 35% underwent revascularization. The vital status of 99% of the patients was determined after a median of 52 months (interquartile range 48 to 55). At follow-up, 88% of patients were taking aspirin, 45% were taking beta blockers, and 50% had undergone revascularization. The survival rate was 70% in patients with > or = 0.5-mm ST segment depression (53%, 77%, and 82% survival for > or = 2-, 1-, and 0.5-mm ST segment depression, respectively; p <0.0001). Patients with a normal electrocardiogram had a greater survival rate (94%) than that of patients with 0.5-mm ST-segment depression (82%, p = 0.020), but not significantly different from that of patients with T-wave inversion (84%, p = NS). Independent predictors of mortality (odds ratio [95% confidence interval]) were: age in yearly increments (1.05 [1.03 to 1.06], p = 0.003), revascularization during follow-up (0.40 [0.29 to 0.56], p = 0.006), pulmonary edema (3.45 [2.19 to 5.45], p = 0.007), and ST-segment depression (1.37 [1.20 to 1.55], p = 0.015). Thus, ST segment depression of > or = 0.5 mm predicts 4-year survival in patients with acute ischemic syndromes. PMID- 10468073 TI - Stent or angioplasty after recanalization of chronic coronary occlusions? (The SARECCO Trial). AB - This study tests whether stent implantation without anticoagulation after catheter recanalization of coronary occlusions can improve outcome compared with balloon angioplasty alone. One hundred ten patients were randomly assigned to angioplasty alone (no stent group) or stent implantation (stent group) after successful recanalization and balloon angioplasty. The type of stent and angioplasty technique utilized were decided by the operator. The acute procedural success in both groups was 100%. The acute minimal lumen diameter (MLD) was 1.85 +/- 0.44 mm in the no stent group versus 2.54 +/- 0.53 mm in the stent group (p <0.01). The diameter stenosis was 21 +/- 13% versus 3 +/- 14% (p <0.01). This was achieved not only by the stent implantation itself but primarily by a larger maximum balloon diameter in the stent group after stent implantation (3.32 +/- 0.55 mm vs 2.86 +/- 0.4 mm, p <0.05). After 4 months, the MLD was 1.15 +/- 0.73 mm in the no stent group versus 1.81 +/- 0.9 mm in the stent group (p <0.01). The diameter stenosis was 56 +/- 29% versus 34 +/- 28% (p <0.01). After 2 years, event-free survival was 26% in the no stent group and 52% in the stent group (p <0.05). Thus, acute and long-term procedural and angiographic success of stent implantation without anticoagulation after recanalization of total coronary occlusions is superior to that of balloon angioplasty alone. This beneficial effect is mainly the result of the larger balloon diameters, which may be used after stent implantation. PMID- 10468074 TI - Time course, magnitude, and consistency of platelet inhibition by abciximab, tirofiban, or eptifibatide in patients with unstable angina pectoris undergoing percutaneous coronary intervention. AB - Adjunctive platelet glycoprotein IIb/IIIa blockade during percutaneous coronary intervention (PCI) reduces platelet-mediated adverse ischemic outcomes. Although abciximab, eptifibatide, and tirofiban have received U.S. Food and Drug Administration approval for use, these agents differ in their pharmacodynamic profiles. Each of these agents has been compared in randomized trials with placebo for patients undergoing PCI, but no randomized comparative studies of these agents have been performed. We compared ex vivo platelet function by both standard light transmission aggregometry and rapid platelet function assay during and after administration of abciximab, eptifibatide, or tirofiban in approved dose regimens on a randomized basis at the time of PCI in patients with unstable angina pectoris. A reduced intensity of platelet inhibition by light transmission aggregometry was observed for tirofiban compared with either eptifibatide or abciximab. In addition, the 30-minute bolus strategy used for tirofiban was associated with delayed onset of maximal platelet inhibition relative to the initiation of bolus infusion. Whether the trends in platelet function observed in this study will be translated into differences in clinical outcomes awaits definition by larger scale randomized clinical trials comparing these platelet glycoprotein IIb/IIIa inhibitors. PMID- 10468075 TI - Gender differences in symptom presentation associated with coronary heart disease. AB - This study explores gender differences in symptom presentation associated with coronary heart disease (CHD). In this prospective study, nurse data collectors directly observed 550 patients as they presented to the Emergency Department (ED) of Yale-New Haven Hospital. The final sample included 217 patients (41% women) diagnosed with CHD (acute coronary ischemia or myocardial infarction). Chest pain was the most frequently reported symptom in women (70%) and men (71%). Unadjusted analyses revealed that women were more likely than men to present with midback pain (odds ratio [OR] 9.61, 95% confidence interval [CI] 2.10 to 44.11, p = 0.001), nausea and/or vomiting (OR 2.29, 95% CI 1.19 to 4.42, p = 0.012), dyspnea (OR 1.82, 95% CI 1.05 to 3.16, p = 0.032), palpitations (OR 3.42, 95% CI 1.02 to 11.47, p = 0.036), and indigestion (OR 2.13, 95% CI 1.03 to 4.44, p = 0.040). After adjustment for age and diabetes, women were more likely to present with nausea and/or vomiting (OR 2.43, 95% CI 1.23 to 4.79, p = 0.011) and indigestion (OR 2.13, 95% CI 1.10 to 4.53, p = 0.048). Women (30%) and men (29%) were equally likely to present without chest pain, and dyspnea was the most common non-chest pain symptom. In the subgroup of patients without chest pain, unadjusted analyses revealed that women were more likely to report nausea and/or vomiting compared with men (OR 4.40, 95% CI 1.30 to 14.84, p = 0.013). Although we found some significant gender differences in non-chest pain symptoms, we conclude that there were more similarities than differences in symptoms in women and men presenting to the ED with symptoms suggestive of CHD who were later diagnosed with CHD. PMID- 10468076 TI - Impact of limited treadmill exercise on adenosine Tc-99m sestamibi single-photon emission computed tomographic myocardial perfusion imaging in coronary artery disease. AB - Limited exercise combined with dipyridamole increases myocardial perfusion defect severity compared with dipyridamole alone. The impact of limited exercise combined with adenosine on myocardial perfusion defect severity is unknown. This study compares myocardial perfusion defect severity with adenosine alone and adenosine combined with limited exercise. Thirty-two patients with coronary artery disease underwent on separate days and in randomized order technetium-99m sestamibi (25 to 30 mCi) single-photon emission computed tomographic imaging at rest, after adenosine (140 microg/kg/min x 6 minutes), and after adenosine (140 microg/kg/min x 4 minutes) during 6 minutes of modified Bruce treadmill exercise (adenosine-exercise). Radiopharmaceutical was injected at 3 and 5 minutes during adenosine and adenosine-exercise, respectively. Images were interpreted by a consensus agreement of 3 nuclear cardiologists without knowledge of patient identity, stress protocol, or clinical data using a 17-segment model and 5-point scoring system. A summed stress score (SSS), summed rest score (SRS), and summed difference (SSS-SRS) score (SDS) were calculated for each image. Peak stress heart rate and rate-pressure product were higher for adenosine-exercise than adenosine (102 +/- 19 vs 81 +/- 11 beats/min and 13,972 +/- 4,265 vs 10,623 +/- 2,131, respectively; both p <0.001). Sensitivity for detection of > or = 50% coronary stenosis was 75% and 72% for adenosine-exercise and adenosine, respectively (p = NS). There were no differences in SSS and SDS between adenosine exercise and adenosine (8.2 +/- 5.9 vs 8.1 +/- 6.3 and 4.9 +/- 4.1 vs 5.2 +/- 4.6, respectively; both p = NS). Thus, in patients with coronary artery disease, limited treadmill exercise combined with adenosine does not increase myocardial perfusion defect severity compared with standard adenosine technetium-99m sestamibi single-photon emission computed tomographic myocardial perfusion imaging. PMID- 10468077 TI - Aspirin absorption rates and platelet inhibition times with 325-mg buffered aspirin tablets (chewed or swallowed intact) and with buffered aspirin solution. AB - Large clinical trials such as the second International Study of Infarct Survival routinely gave patients with myocardial infarction a chewed aspirin, yet there are no data to show whether chewing of aspirin is better, or worse, than swallowing a whole tablet. We performed a randomized, placebo-controlled study to determine whether chewing aspirin or administering it in solution accelerates its absorption and antiplatelet activity. On separate days, 12 fasting volunteers ingested 325 mg of buffered aspirin, either by chewing a tablet for 30 seconds before swallowing it with 4 ounces of water, swallowing a whole tablet with 4 ounces of water, or drinking 4 ounces of Alka Seltzer. Frequent blood samples were obtained for serum aspirin, salicylate, and thromboxane B2 (TxB2) concentrations. With all formulations of aspirin, serum TxB2 decreased 50% when the plasma aspirin concentration reached approximately 1,000 ng/ml. A 50% and 90% decrease in serum TxB2 occurred more quickly after chewing a tablet than after a tablet was swallowed whole. For example, the t 50% for serum TxB2 inhibition was 5.0 +/- 0.6 minutes with the chewed tablet versus 12.0 +/- 2.3 minutes when the tablet was swallowed (p = 0.01). A 50% decrease in serum TxB2 occurred 7.6 +/- 1.2 minutes after Alka Seltzer solution (p = 0.04 vs chewing a tablet; p = 0.13 vs swallowing a whole tablet). Chewing an aspirin tablet is the most effective way of accelerating absorption of aspirin into the blood and shortening the time required for an antiplatelet effect. PMID- 10468078 TI - Quantitative angiographic analysis of stent restenosis in the Scripps Coronary Radiation to Inhibit Intimal Proliferation Post Stenting (SCRIPPS) Trial. AB - To identify luminal dimension changes occurring within the stent alone and within the stent + margin segment, we reviewed the quantitative angiographic results obtained from the Scripps Coronary Radiation to Inhibit Proliferation Post Stenting (SCRIPPS) trial, a prospective randomized trial assessing the effect of iridium-192 (Ir-192) on the prevention of stent restenosis. Fifty-five patients were randomly assigned to receive Ir-192 or placebo sources after successful intervention. Procedural and 6-month follow-up cineangiograms were quantitatively reviewed in 52 patients to identify changes within the stent and the stent + margin segment. The percent diameter stenosis was lower within the stent than within the stent + margin segment after the procedure (6 +/- 22% vs 21+/- 15%, p <0.0001) and at follow-up (28 +/- 29% vs 42 +/- 21%, p <0.0001). As a result, a lower restenosis rate was found within the stent than within the stent + margin (25% vs 37%, p <0.0001); isolated stent margin restenosis occurred in 11.5% of lesions. Treatment with Ir-192 reduced restenosis within the stent (8% vs 39%; p = 0.010) and within the stent + margin segment (17% vs 54%; p = 0.010); the reduction in restenosis at the margin only (8.3% vs 14.3%, p = 0.503) was not significant. The lowest relative risk for restenosis resulting from Ir-192 occurred within the stent (0.21; 95% confidence interval [CI] 0.05 to 0.86) compared with the stent + margin segment (0.31; 95% CI 0.12 to 0.81) or the stent margin (0.58; 95% CI 0.12 to 2.91). In the SCRIPPS trial, 32% of restenosis occurred at the stent margins. Treatment with Ir-192 reduced restenosis primarily within the stent rather than the margin. Whether extending the treatment length to fully include the stent margins will further reduce restenosis requires further study. PMID- 10468079 TI - Early temporal changes in coronary flow velocity patterns in patients with acute myocardial infarction demonstrating the "no-reflow" phenomenon. AB - Coronary flow velocity pattern in patients with acute myocardial infarction demonstrating no-reflow phenomenon is characterized with early systolic retrograde flow and rapid deceleration of diastolic flow velocity. In this study, we investigated the early temporal changes in microvascular function in patients with the no-reflow phenomenon. Among 144 patients with a first acute myocardial infarction, 33 exhibited sizable no-reflow phenomenon after coronary reperfusion with myocardial contrast echocardiography. We assessed temporal changes in coronary flow velocity patterns with the Doppler guidewire. The early systolic retrograde flow was observed < or = 10 seconds after reperfusion in 16 patients (group A) or later in 17 patients (331 +/- 327 seconds, group B). Diastolic deceleration rate was higher in group A than in group B at 1 minute after reperfusion. It gradually increased in group B and showed comparable value to group A 10 minutes later. Group A had longer elapsed time from symptom onset to reperfusion and a greater number of infarct Q waves before reperfusion than group B (14 +/- 13 vs 5 +/- 2 hours, p <0.01; and 3 +/- 2 vs 2 +/- 1, p <0.02). In contrast, the incidence of transient ST reelevation shortly after reperfusion was higher in group B (76% vs 25%, p <0.01). Thus, the characteristic coronary flow velocity pattern is either established at the moment of coronary reperfusion or progresses thereafter in patients with no-reflow phenomenon. This suggests different mechanisms of developing ischemic microvascular injury. PMID- 10468080 TI - Effect of radiofrequency ablation on atrial mechanical function in patients with atrial flutter. AB - Atrial stunning, as assessed by left atrial appendage emptying and increased spontaneous echo contrast, is known to occur following direct-current cardioversion of atrial fibrillation (AF) and atrial flutter (AFI). Little is known on atrial mechanical function and the time course of atrial recovery following radiofrequency ablation of AFI. Fourteen patients undergoing radiofrequency ablation of persistent typical counterclockwise AFI were enrolled. Two-dimensional and pulse Doppler transesophageal echocardiography (TEE) were performed before ablation and immediately following restoration of sinus rhythm. Left atrial spontaneous echo contrast grades, left atrial appendage emptying fractions, and peak left atrial appendage emptying velocities were measured. Transthoracic echocardiography (TTE) was performed immediately after ablation, then repeated after 1 day, 1 week, and 6 weeks to measure peak transmitral velocities and percent atrial contribution to ventricular filling. Left atrial appendage emptying velocities decreased significantly following AFI termination (44 +/- 23 cm/s before ablation vs 25 +/- 14 cm/s after ablation, p = 0.01). Left atrial appendage emptying fractions also decreased significantly (0.48 +/- 0.1 preablation vs 0.34 +/- 0.17 postablation, p = 0.02). New spontaneous echo contrast developed in 4 patients (29%) after ablation. Four patients had complete atrial standstill after ablation, and 1 patient developed a new left atrial appendage thrombus. The percent atrial contribution to ventricular filling recovered progressively over 6 weeks with significant improvement in peak transmitral velocities at day 7. Thus, atrial stunning occurs after catheter ablation of AFI and may lead to rapid formation of thrombus in the left atrial appendage. Significant improvement in left atrial function occurs in 7 days. PMID- 10468081 TI - Evaluation of biatrial pacing, right atrial pacing, and no pacing in patients with drug refractory atrial fibrillation. AB - It has been suggested biatrial pacing may prevent the recurrence of atrial fibrillation (AF). To further evaluate this hypothesis, we performed a randomized, single-blinded study in 19 patients with drug refractory AF. The study compared biatrial pacing with conventional right atrial (RA) pacing and a control period of inhibited pacing. The pacing modes utilized were DDD with a base rate of 70 beats/min for biatrial and RA pace (with and without biatrial resynchronization, respectively) and 40 beats/min for the control period. The duration of each pacing mode was 3 months. The number of AF episodes and their duration were obtained from pacemaker Holter memory (Chorus RM ELA Medical). Comparison of the control period (n = 11) with either pacing strategy showed a significant decrease in the total duration of AF (control 27 +/- 35 days, biatrial 8 +/- 15 days p = 0.02, RA 11 +/- 27 days p = 0.04). However, there was no effect on the number of AF episodes (control 79 +/- 108, biatrial 36 +/- 75 p = 0.32, RA 41 +/- 80 p = 0.11). The total percentage of atrial pacing also significantly increased when the control period (6 +/- 9%) was compared with both RA pace (62 +/- 33%, p = 0.008) and biatrial pace (63 +/- 31, p = 0.003). When biatrial pacing was compared with RA pace (n = 19), there was no significant difference in either the duration of AF (biatrial 16 +/- 26 days vs RA 19 +/- 31 days, p = 0.7) or the number of AF episodes (biatrial 56 +/- 91 vs RA 87 +/- 106, p = 0.34). In conclusion, pacing (either type) at a base rate of 70 beats/min has an antifibrillatory effect when compared with inhibited pacing at 40 beats/min. No additional benefit of biatrial pacing over right atrial pacing was demonstrated in this study. PMID- 10468082 TI - Evaluation of growth hormone administration in patients with chronic heart failure secondary to coronary artery disease. AB - We have examined the effects of 6 months of treatment with growth hormone (GH) (0.02 U/kg/day) in 10 patients with chronic postischemic cardiac failure. Ten patients matched for age, body mass index, functional class, and ejection fraction served as a control group. In the GH group, 1 patient died and 2 were withdrawn from the study because of arrhythmia or worsening of heart failure. In the control group, 1 patient died and 1 patient was withdrawn from the study because of progressive heart failure. Among GH patients, those with an unfavorable outcome had a greater left ventricular end-diastolic diameter (79, 82, and 88 mm) on entry to the study than patients without adverse events (range 62 to 72 mm). At the end of the study, the seven GH patients reported a feeling of well-being and had a significant increase in their exercise test duration (462 +/- 121 vs 591 +/- 105 seconds, p <0.05). Low baseline insulin-like growth factor I values were increased with GH treatment (189 +/- 52 vs 100 +/- 22 ng/ml, p <0.01). GH did not change left ventricular diameters or wall thickness. A trend toward decreased serum triglyceride levels and adipose body tissue associated with an increase in high-density lipoproteins was observed in the GH group. In conclusion, our present data support previous suggestions that GH treatment exerts some beneficial effects in patients with chronic, stabilized, moderately severe heart failure, but may have deleterious effects in patients with more severe heart failure. PMID- 10468083 TI - Prognostic utility of myocardial blood flow assessed by N-13 ammonia positron emission tomography in patients with idiopathic dilated cardiomyopathy. AB - Previous studies in patients with idiopathic dilated cardiomyopathy (IDC) have suggested that myocardial perfusion is impaired and spatially heterogeneous in such cases. Our objective was to identify any association between an abnormality in myocardial perfusion and the prognosis of patients with IDC. We collected data on N-13 ammonia positron emission tomography (PET) studies performed in 26 patients with IDC (9 nonsurvivors, 17 survivors) and in 8 normal control subjects. Regional myocardial blood flow (rMBF) was quantified using N-13 ammonia positron emission tomography and the Simple flow model. The spatial heterogeneity of myocardial perfusion was assessed by calculating the coefficient of variance of rMBF. Mean rMBF of the survivors was significantly lower (0.54 +/- 0.13 ml/min/g) than that of control subjects (0.66 +/- 0.06 ml/min/g) (p = 0.03 vs control), but did not differ significantly between nonsurvivors (0.58 +/- 0.15 ml/min/g) and control subjects. The coefficient of variance of rMBF was significantly higher in nonsurvivors than in either survivors or control subjects (0.24 +/- 0.08 vs 0.15 +/- 0.08, p = 0.007, and 0.16 +/- 0.05, p = 0.03, respectively). The probability of 3-year survival (Kaplan-Meier method) was 33.0% in subjects whose coefficient of variance of rMBF was above the median compared with 90.0% in subjects whose coefficient of variance of rMBF was below the median (p = 0.01). The probability of 3-year survival did not differ among subjects whose mean rMBF was above versus below the median (61.5% vs 62.9%, respectively). The results suggest that the prognosis of patients with IDC is associated with the spatial heterogeneity of myocardial perfusion, not with initial absolute rMBF. PMID- 10468084 TI - Effects of age and physiologic variables on right ventricular filling dynamics in normal subjects. AB - The reference values for right ventricular (RV) filling of normal persons and the effects of physiologic variables in a large series have not been described. The objective of this study was to characterize superior vena cava, hepatic vein, and RV inflow Doppler measurements in a large normal reference group to reflect the aging process, gender, heart rate, and effects of respiration. We prospectively performed pulsed-wave Doppler echocardiography of the superior vena cava, hepatic vein, and RV inflow during inspiration, expiration, and apnea in 115 healthy volunteers (62 women and 53 men) ranging in age from 21 to 84 years (mean +/- SEM 48 +/- 17). For analysis, the study subjects were classified by age into 2 groups: those < 50 years of age (group 1; n = 60) and those > or = 50 years of age (group 2; n = 55). Multiregression models were used to assess the influence of age, gender, and heart rate on Doppler variables. There were important differences in superior vena cava and RV inflow between the 2 groups. Group 2 had a greater superior vena cava peak atrial flow velocity (16 +/- 3 vs 13 +/- 3 cm/s), flow integrals (1.5 +/- 0.4 vs 1.1 +/- 0.3 cm), and reverse flow as a percentage of forward flow (17 +/- 6% vs 14 +/- 6%) than group 1. In group 2, peak RV inflow early filling velocity (41 +/- 8 vs 51 +/- 7 cm/s) and ratio of early filling-to-atrial filling (1.3 +/- 0.4 vs 2 +/- 0.5) were lower than that of group 1. Likewise, peak atrial filling velocity was higher (33 +/- 8 vs 27 +/- 8 cm/s) and deceleration time was longer (198 +/- 23 vs 188 +/- 22 ms) in group 2. The superior vena cava and hepatic vein peak forward flow velocities were significantly higher during inspiration than during expiration and apnea. Similarly, RV inflow velocities were significantly higher during inspiration than in expiration and apnea. Multiregression analysis showed that age, gender, and heart rate had important effects on Doppler variables. Thus, this study demonstrates the effects of aging and normal physiologic variable flow velocities in the superior vena cava, hepatic veins, and RV inflow in a large series of normal subjects. PMID- 10468085 TI - Circadian patterns of heart rate variability, fibrinolytic activity, and hemostatic factors in type I diabetes mellitus with cardiac autonomic neuropathy. AB - Diabetes mellitus is associated with a marked increase in the risk of coronary events but with an altered circadian distribution that demonstrates an absent morning peak and higher infarction rate during the evening hours. To elucidate the mechanism of this phenomenon, the circadian pattern of heart rate variability was evaluated in 22 type I diabetic patients with diabetic autonomic neuropathy in conjunction with circadian changes of fibrinolytic and hemostatic factors. The circadian pattern (6 A.M. to 10 P.M. vs 10 P.M. to 6 A.M.) of 3 indexes of parasympathetic tone was evaluated using 24-hour heart rate variability analysis. The high-frequency power (3.0 +/- 0.2 vs 3.3 +/- 0.2 ms2, p = 0.08) and the percentage of RR intervals with >50 ms variation (0.47 +/- 0.18 vs 0.69 +/- 0.33 ms, p = 0.52) demonstrated no significant circadian variation. The square root of mean squared differences of successive RR intervals showed a small but significant increase during nighttime (8.5 +/- 0.7 vs 9.7 +/- 1.1 ms, p = 0.02). Fibrinolytic activity was significantly lower at 8 A.M. than at 4 P.M. (166.4 +/- 12.5 to 200.2 +/- 9.3 mm2, p = 0.0003), but with a low amplitude. Plasminogen activator inhibitor 1 showed no circadian variation. Factor VII and fibrinogen demonstrated a significant reduction from 8 A.M. to 4 P.M., but both peak and nadir values were elevated. The von Willebrand factor was markedly elevated with no circadian variation. Thus, diabetic autonomic neuropathy is associated with a loss of both the nocturnal predominance of parasympathetic activity and a prothrombotic state that persists throughout the day. These abnormalities may attenuate the relative protection from coronary events during the afternoon and nighttime. PMID- 10468086 TI - Future cardiovascular end point studies: where will the research take us? AB - Coronary artery disease is the most common cause of death in the world. Emerging concepts of atherosclerosis imply that atherosclerosis is a diffuse disease, and cannot be definitively treated with local, anatomic interventions, such as coronary artery bypass graft surgery or angioplasty. Cholesterol lowering, on the other hand, has been shown to dramatically lower the rate of both morbid and mortal coronary events. In trials with new statin drugs, coronary risk has been lowered by approximately 30%. Additional risk reduction will require other approaches, including (1) intervention for other risk factors, (2) more aggressive cholesterol lowering, or (3) increased attention to primary prevention. The last requires a combination of public health measures to change harmful diet and life-style patterns as well as case findings to identify and treat at-risk subjects. For all these approaches, measures that will increase compliance by both physicians and patients to regimens with proven benefits are required. PMID- 10468087 TI - Elevated levels of C-reactive protein before coronary artery bypass grafting predict recurrence of ischemic events. AB - C-reactive protein was measured in 86 patients undergoing coronary artery bypass graft surgery. Patients were followed up for 3.2 years (range 1 to 6). Patients with C-reactive protein > or = 3 mg/L had significantly increased risk of recurrent ischemia at 1 to 6 years after intervention. PMID- 10468088 TI - Histopathologic comparison of human coronary in-stent and post-balloon angioplasty restenotic tissue. AB - Histomorphometric studies for smooth muscle cells, macrophages, tissue factor, collagen, and cell proliferation were performed on 20 atherectomy specimens from patients with in-stent restenosis and 20 specimens from patients with post balloon PTCA restenosis. Smooth muscle cell content was larger and proliferation index was higher in in-stent restenotic tissue; macrophage, tissue factor, and collagen content were larger in post-balloon percutaneous transluminal coronary angioplasty restenotic tissue, suggesting a more cellular and proliferative response with less thrombogenic potential in human tissue from in-stent restenosis than tissue from post-balloon percutaneous transluminal coronary angioplasty restenosis. PMID- 10468089 TI - Coronary arterial involvement and QT dispersion in Kawasaki disease. AB - For the early detection of myocardial ischemia in patients with severe involvement of the coronary arteries after Kawasaki disease, a method with high sensitivity and low cost is desirable because these patients require frequent follow-up and diagnostic tests. For this purpose, electrocardiographic, echocardiographic, Holter, and stress testing or angiography are repeated. However, these tests have some limitations due to cost, convenience, or sensitivity. It is uncertain that increased QT dispersion would exactly indicate progression of myocardial ischemia after Kawasaki disease, but this is the first study to present that QT dispersion of > or = 60 ms had higher sensitivity for detection of severe involvement of coronary artery after Kawasaki disease. This study is limited due to the small number of patients; larger prospective studies are required to clarify the usefulness of QT dispersion analysis in detecting the progression of myocardial ischemia after Kawasaki disease. PMID- 10468090 TI - Association of left ventricular hypertrophy and chronic atrial fibrillation with the incidence of new thromboembolic stroke in 2,384 older persons. AB - In a prospective study of 2,384 persons, mean age 81 years, at 44-month follow up, new thromboembolic stroke developed in 510 of 2,384 persons (21%). The Cox regression model showed that significant independent risk factors for new thromboembolic stroke were atrial fibrillation (risk ratio 3.2), left ventricular hypertrophy (risk ratio 2.8), prior stroke (risk ratio 2.2), and male gender (risk ratio 1.2). PMID- 10468091 TI - Cardiac troponin I and cardiac enzymes after electrophysiologic studies, ablations, and defibrillator implantations. AB - Diagnostic electrophysiologic studies and defibrillator implantations with subsequent test discharges from the defibrillator do not cause an elevation in cardiac troponin I levels. All patients with an ablation had an elevation in the cardiac troponin I levels. PMID- 10468092 TI - Quality of life before and after radiofrequency catheter ablation in patients with drug refractory atrioventricular nodal reentrant tachycardia. AB - In a retrospective survey of 161 highly symptomatic patients, we found significant improvements in symptoms, patient utility, and use of medical care services after radiofrequency ablation for atrioventricular nodal reentrant tachycardia. PMID- 10468093 TI - Transiliac permanent pacemaker implantation after extraction of infected pectoral pacemaker systems. AB - Infected pectoral pacemaker systems were extracted in 5 patients and new pacemakers were reimplanted in a pelvic pocket through the iliac veins. In patients who have infected pectoral pacemaker systems, this easy and safe technique provides an alternate route for reimplantation of permanent pacemakers. PMID- 10468094 TI - Effect of vasoactive therapy on heart rate variability in patients with acutely decompensated congestive heart failure. AB - Heart rate variability is markedly reduced in patients with acutely decompensated heart failure. Improvement in heart rate variability is closely associated with improvement in heart failure, which may represent a decrease in sympathetic overactivity. PMID- 10468095 TI - Use of enoxaparin for the chronically anticoagulated patient before and after procedures. AB - Patients who require chronic anticoagulation and a procedure have been traditionally managed either by stopping warfarin and starting intravenous standard heparin or by adjusted dose subcutaneous standard heparin or taken off all anticoagulation for a week before the procedure. Enoxaparin may be useful as an alternative method of anticoagulation, avoiding hospitalization and the need for frequent monitoring. PMID- 10468096 TI - Frequency and prognostic significance of pericardial effusion in primary pulmonary hypertension. PPH Study Group. Primary pulmonary hypertension. AB - Pericardial effusion was noted in 43 of 79 patients (54%) with severe primary pulmonary hypertension. Larger effusion was associated with hemodynamic and echocardiographic evidence of right heart failure, impaired exercise tolerance, and a poor 1-year prognosis. PMID- 10468097 TI - Simultaneous hypertrophic cardiomyopathy and ventricular septal defect in children. AB - Our findings are based on clinical observations and not on a designed study. However, in our University Hospital area, all children with suspected or verified HC are sent to the same pediatric cardiologist (TT). Therefore, our patients are probably representative of children with HC, at least in East Finland. Although the number of patients was small, the prevalence of VSD in children with HC was so high, that it is improbable that the association between the 2 diseases is due to chance alone. Larger studies are needed to confirm our results and to show if muscular VSDs are typical of all HC cases or of specific subgroups only. We suggest that children with HC should undergo a thorough echocardiographic examination to detect possible muscular VSDs, which are usually readily detected by modern color flow mapping techniques. Importantly, in children with a muscular VSD, the possibility of HC as an underlying pathology should be kept in mind. PMID- 10468098 TI - Two versus six hours of bed rest following left-sided cardiac catheterization and a meta-analysis of early ambulation trials. AB - The present study and associated meta-analysis shows that 6 hours of bed rest after outpatient cardiac catheterization offers no advantage over 2 hours of bed rest with respect to groin bleeding complications. The occasional bleeding observed occurred shortly after ambulation rather than after discharge. PMID- 10468099 TI - Cardiac observations late after operative transmyocardial laser "revascularization". AB - This case study suggests that transmyocardial laser revascularization does not produce lasting benefit. PMID- 10468100 TI - Congenital aneurysm of the descending aorta in a neonate. AB - A large aneurysm of the descending aorta of unknown etiology was diagnosed in utero. Medical treatment with propranolol was continued after surgery which was performed at the age of 2 months. PMID- 10468101 TI - Heart rate variability after coronary artery bypass grafting. PMID- 10468102 TI - Dual hepatic metabolism of cerivastatin--clarifications. PMID- 10468103 TI - Provocative tests for coronary artery spasm. PMID- 10468104 TI - Effects of thirty-four adolescent tobacco use cessation and prevention trials on regular users of tobacco products. AB - Since 1991, adolescent tobacco use rates have increased while adult use has steadily decreased. The failure of adolescent tobacco use cessation and prevention programs to reduce this overall smoking rate indicates that research must be advanced in this area. As a start, the current status of cessation and prevention research that targets adolescent regular tobacco users should be stated. This paper contributes to that goal by reviewing the last two and a half decades of research in this area. A total of 34 programs, equally divided between cessation and prevention (targeting regular tobacco users), are presented and relevant data are provided for each. Among the cessation studies, an emphasis of programming on immediate consequences of use, and instruction in coping strategies, may have led to relatively successful programs. Prevention studies arguably may have achieved lower success rates but were applied to a larger sample with a longer follow-up period. Despite showing some success, it is apparent that the scientific status of cessation research is less refined than prevention research. More research is needed to define the most successful approaches for cessation of adolescent tobacco use. PMID- 10468105 TI - Addiction careers and careers in addiction. AB - Research into resocialization of criminals or into recovery from substance dependence has stressed the need to give up the deviant identity and lifestyle. However, addiction careers, crime, and other types of deviant behavior require a wide range of skills. Such competence may facilitate the alternative or simultaneous pursuit of "respectable" careers in treatment, prevention, research, or policy matters. Former alcoholics fill important positions in the service structures of Alcoholics Anonymous, current or former users are appreciated as partners in AIDS prevention programs, and drug addicts can become privileged access interviewers and be recruited by research institutes. The paper discusses "market conditions" that favor or impede career shifts. How society and professionals perceive individuals with a history of stigma varies from complete rejection to admiration or recognition of usefulness. This assessment depends on such factors as degree of political and scientific interest in controlling, changing, and detecting hidden deviant populations, as well as public fascination with "authentic" deviants, combined with increasing scepticism about conventional expert knowledge. PMID- 10468106 TI - Cocaine--profiles, drug histories, and patterns of use of patients from Brazil. AB - We studied profiles and drug histories of 294 cocaine users from 15 treatment services in Sao Paulo, Brazil during 1996-1997. Mean age of subjects was 27 years, and 90% were male. Over 50% had used five different substances apart from cocaine, usually tobacco, alcohol, cannabis, tranquilizers, and solvents. Mean age at first cocaine use was 18.9 years by which time 87% had snorted the drug. Thirty-two percent had injected cocaine, 82% had smoked crack, and 74% reported a full route transition. Sixty-three percent reported daily cocaine use. Median duration of cocaine use was 6.3 years. Acts of acquisitive crime were common, and 56% had been arrested. Our finding are discussed in terms of implications for prevention and treatment. PMID- 10468107 TI - The Helping Alliance Questionnaire: psychometric properties in patients with substance dependence. AB - Psychometric properties of the Helping Alliance Questionnaire (HAQ) are analyzed in a population of 340 substance-dependent patients of an addiction clinic in the Netherlands. Factor analysis yields a two-factor structure: Cooperation and Helpfulness. The scales show fair correlations with three out of seven scales from the Barrett-Lennard Relationship Inventory. Length of Stay in Detox is predicted by scores on the Helpfulness scale and noncompliance by scores on the Cooperation scale. Besides this, intermediate outcome measures are correlated with HAQ scores. The HAQ seems to be a "quick scan" instrument to give a quick and global impression of the patients' perception of the quality of the working alliance with the therapist. PMID- 10468108 TI - A preliminary evaluation of the effectiveness of a standardized Student Assistance Program. AB - We examined the effectiveness of Student Assistance Program group counseling. One hundred and forty-four (144) substance using high school seniors completed a questionnaire in 1995 concerning their substance use before and after counseling, their experience in counseling, and the impact of counseling on their substance use. Participants reported significant decreases in substance use and attributed these changes to counseling. Most participants liked counseling, and higher frequency drinkers did not differ from lower frequency drinkers in liking counseling and reporting decreased substance use. These results offer support for the effectiveness of such interventions and highlight the need for additional research in the area. PMID- 10468109 TI - Agreement between urinalysis and self-reported drug use. AB - One hundred fifty-eight drug users received an interview that included self reported drug use. Opiate/cocaine use in the prior 48 hours was assessed by urinalysis. Fifty-five subjects received a urine test after the interview and 103 were tested before. Chi-square tests were performed to determine if agreement between self-reported drug use and urine test results was associated with timing of urine testing. The rate of agreement was 58% when the urine test was performed after the interview and 93% when performed before the interview (chi2 = 28.6, p < .001). Conducting urine tests before an interview can increase the accuracy of self-reports. PMID- 10468110 TI - Cognitive psychopathology of problem gambling. AB - Drawing on the available research literature on the cognitive distortions present during gambling, a typology of gambling-relevant cognitive distortions is presented. These include the magnification of gambling skills, minimization of other gambler's skills, superstitious beliefs (including talismanic, behavioral, and cognitive superstitions), interpretive biases (including internal attributions, external attributions, gambler's fallacy, chasing, anthropomorphism, reframed losses, hindsight bias), temporal telescoping, selective memory, predictive skill, illusion of control over luck (including luck as an uncontrollable variable, luck as a controllable variable, luck as a trait variable, luck as a contagion), and illusory correlation. In addition, gambling relevant beliefs about the self are also discussed, including entitlement, omnipotence, cognitive reasoning, and magical thinking. PMID- 10468111 TI - Selenium deficiency and chronic pancreatitis: disease mechanism and potential for therapy. AB - BACKGROUND: It has been suggested that antioxidant deficiency may play a role in the pathogenesis of chronic pancreatitis. The aim of this review was to analyse the evidence for this relationship and to consider the role of antioxidant supplementation in the treatment of chronic pancreatitis. METHODS: Medline review of all English language publications for the years 1966-1998. RESULTS AND CONCLUSIONS: There is evidence that patients with chronic pancreatitis have enhanced levels of free radical production, cytochrome P450 induction and antioxidant deficiencies, in particular selenium. The limited published literature in this field suggests that dietary antioxidant supplementation may ameliorate the pain associated with chronic pancreatitis, diminish the frequency of acute exacerbations and reduce the requirement for pancreatic surgery. These findings await confirmation by a large prospective placebo-controlled study. PMID- 10468112 TI - What causes cancer gallbladder?: a review. AB - Gallbladder cancer is a common malignancy of the biliary tract. It is the fifth common malignancy of the gastrointestinal tract in United States and third in Northern India. Despite such high prevalence, there is scanty published literature about this disease in indexed journals. Therefore, this article is intended to provide a brief overview of gallbladder cancer risk factors, based mainly on published evidence from analytical epidemiology and recent research findings of biologists and practising oncologists. Furthermore, an attempt has been made to establish an association between different causative factors and the occurrence of the disease. PMID- 10468113 TI - The need to handicap the recipient's native liver in the rat model of heterotopic auxiliary liver transplantation. AB - In the rat model of heterotopic auxiliary liver transplantation (HALTx), the opinion varies on whether and how the recipient's native liver should be handicapped. To avoid atrophy of the transplanted organ, in this study, two different handicaps were evaluated and their effects on post-operative animal survival and liver biology are described. With a sole portacaval shunt (group 1) all rats survived longer than 3 months. An additional handicap of the liver with either a 68% partial hepatectomy (68% PH) (group 2), or both a 68% PH and a common bile duct ligation (CBDL) (group 3) led to a 100% mortality within 2 days after surgery. When an auxiliary liver was transplanted to the rats handicapped with a 68% PH (group 4), serum Bilirubin and ALAT values were significantly lower than those handicapped with both a 68% PH and a CBDL (group 5). Autopsy and histology of the long-term survivors revealed the atrophy of the engrafted livers and the regeneration of the native livers in group 4, whereas it showed the opposite in group 5. Thus the various manipulations of the native liver do influence differently the post-transplant animal survival, serum liver biochemistry and the outcome of the engrafted liver in this rat model of HALTx. PMID- 10468114 TI - Results of a modified sugiura's devascularisation in the management of "unshuntable" portal hypertension. AB - The results of a modified Sugiura devascularisation procedure were assessed in 14 patients with thrombosis of the portal and splenic vein requiring surgery for variceal hemorrhage, with no vein suitable for orthodox shunt surgery. The venous anatomy was determined by ultrasonography with Doppler studies and portovenography. Liver biochemistry as well as liver architecture on histopathology was normal in all. The surgery was elective in 9 cases for documented bleed from diffuse fundal gastric varices (FGV) and emergency in 5 cases, 3 having bleeding FGV and 2 for failure of emergency esophageal variceal sclerotherapy. All were subjected to a transabdominal extensive devascularisation of the upper two third of the stomach and lower 7-10cm of the esophagus. Stapled esophageal transection (n = 11) or esophageal variceal underrunning (n = 1) was performed in all with esophageal varices. FGV were underrun. Follow up endoscopies were done six monthly. There were 9 males and 5 females with a mean age of 17.2 years (SD 12.8). There was no operative mortality. Acute variceal bleeding was controlled in all patients. Over a mean follow up of 38 months, all but one remain free of recurrent bleeding. We conclude that a modified Sugiura devascularisation procedure is effective in the immediate and medium term control of variceal bleeding in patients with "unshuntable" portal hypertension. PMID- 10468115 TI - Bilirubin as an anti-oxidant for surgical stress: a preliminary report of bilirubin oxidative metabolites. AB - BACKGROUND: Bilirubin has been recognized as an anti-oxidant. The purpose of this study was to examine whether bilirubin would act as an antioxidant for surgical stress in humans. MATERIALS AND METHODS: Serum bilirubin and urinary bilirubin oxidative metabolites (BOM) were measured in 96 patients who underwent surgery. The antioxidant activity of bilirubin was assessed using BOM measured by enzyme linked immunosorbent assay with an anti-bilirubin monoclonal antibody. RESULTS: Serum bilirubin levels increased after surgery in all 96 patients (p < 0.01), but did not correlate with operation time or blood loss (p = 0.53 and p = 0.28, respectively). BOM increased only in patients with major surgeries (p = 0.048). Significant correlations between BOM and operation time and blood loss were found (p < 0.01). CONCLUSIONS: Bilirubin appears to act as an antioxidant for invasive surgery in humans. Urinary BOM could be a reliable marker for the degree of surgical stress. PMID- 10468116 TI - The real incidence of extracapsular (satellite) cysts of liver echinococcus. AB - BACKGROUND: The presence of extracapsular (Satellite) cysts in liver echinococcus granulosus is known for many years. In one of our previous studies of radiological (CT) material they were found to be present in 16% of cases. METHODS: In the present study the operative findings, in cases of total capsectomy (cystopericystectomy) or partial lobectomy are presented. RESULTS: The real incidence of these cysts in operative specimens was as high as 29,5%. They were present in 15 out of 51 totally excised cysts. CONCLUSIONS: We conclude that satellite cysts are present more often than they are radiologically detected. As they can be incriminated as a cause of recurrence of the disease they must be excised en block with the main parasitic cysts, by means of more radical procedures such as cystopericystectomy or partial hepatectomy, whenever it is feasible. PMID- 10468117 TI - Surgical management of calcified hydatid cysts of the liver. AB - Hydatid disease of the liver is still a major cause of morbidity in Greece. Beside the common complications of rupture and suppuration, calcification of the hepatic cysts represent a not well studied, less frequent and sometimes difficult surgical problem. In the present study 75 cases with calcified symptomatic liver echinococcosis were operated on in the 1st Propedeutic Surgical Clinic between 1964 to 1996. Twenty-eight patients were male and 47 female with ages from 23 to 78 years. The diagnosis was based mainly on the clinical picture and radiological studies. In 5 cases the operative method was cystopericystectomy. We performed evacuation of the cystic cavity and partial pericystectomy and primary closure of the residual cavity in 6 cases, omentoplasty or filling of the residual cavity with a piece of muscle of the diaphragm in 4 cases and external drainage by closed tube, in 60 cases. In 12 of those with drainage, after a period of time, a second operation with easy, removal of most of the calcareous wall plaques was performed. The mortality rate was 2%. Our results could be considered satisfactory. In the calcified parasitic cysts of the liver the proposed technique is cystopericystectomy. An alternative procedure is pericystectomy and drainage with a "planned" reoperation with a bloodless, due to intervening inflammation, chiseling of the calcification. PMID- 10468118 TI - Liver resections for metastases from intraabdominal leiomyosarcoma. AB - This paper discusses liver resection for intraabdominal leiomyosarcoma metastases as a therapy for carefully selected patients. Of the 83 hepatectomies performed from 1992 to 1996, five were resections for liver metastases due to intraabdominal leiomyosarcoma, in 3 patients. The surgical indication was single liver metastases, without any evidence of extrahepatic disease. No mortality occurred during surgery and the longest survival was 38 months. We concluded that liver resection for leiomyosarcoma metastases can be performed, allowing a long term survival in an occasional patient. PMID- 10468119 TI - Left extended hemihepatectomy with preservation of large inferior right hepatic vein: a case report. AB - For hepatic function to be preserved after an extended hemihepatectomy adequate venous drainage of the remaining liver is required. Most metastases close to the confluence of the superior hepatic veins are considered unresectable because hepatic venous outflow after resection would be compromised. In 10-25% of people, the inferior right hepatic vein is of large calibre. Thus the superior hepatic veins may be sacrificed and hepatic function preserved if a large inferior right hepatic vein is present. A patient with involvement of segments 2, 4 and 8 by metastatic colorectal cancer is presented. This patient had a large inferior right hepatic vein, and so was able to undergo an extended left hemihepatectomy with ligation of all superior hepatic veins. Subsequent quality of life was maintained. This case illustrates that an 'unresectable' hepatic lesion can be actually resectable if an alternative venous drainage is present. A pre-operative search for a prominent inferior right hepatic vein by ultrasound, computerised tomography, or even magnetic resonance imaging should be considered in these cases. PMID- 10468120 TI - Monolobar Caroli's Disease and cholangiocarcinoma. AB - Caroli's Disease (CD) is a rare congenital disorder characterized by cystic dilatation of the intrahepatic bile ducts. This report describes a patient with cholangiocarcinoma arising in the setting of monolobar CD. In spite of detailed investigations including biliary enteric bypass and endoscopic retrograde cholangiography, the diagnosis of mucinous cholangiocarcinoma (CCA) was not made for almost one year. The presentation, diagnosis and treatment of monolobar CD and the association between monolobar CD and biliary tract cancer are discussed. Hepatic resection is the treatment of choice for monolobar CD. PMID- 10468121 TI - Arterialisation of the portal vein with an aortoportal jump graft for portal vein thrombosis following liver resection for malignancy. AB - Fibrolamellar hepatocellular carcinoma (FHCC) is a variant of hepatocellular carcinoma, which mainly affects a young age group and carries a relatively good prognosis. It is widely accepted that aggressive curative resection is still the best option for FHCC. We report here a case of successful arterialisation of the portal vein with an aortoportal jump graft for portal vein thrombosis, which developed postoperatively in an already comprised portal vein with tumour invasion following an extensive liver resection for FHCC. PMID- 10468122 TI - Consensus statement on diagnosis, investigation, treatment and prevention of acute bacterial meningitis in immunocompetent adults. British Infection Society Working Party. PMID- 10468123 TI - Lipid amphotericin B combinations; 'la creme de la creme'? PMID- 10468124 TI - Nucleic acid amplification for mycobacterial diagnosis. PMID- 10468125 TI - Gram-positive endocarditis. PMID- 10468126 TI - AIDS-associated cryptococcal meningitis in Rwanda (1983-1992): epidemiologic and diagnostic features. AB - OBJECTIVES: to document the trend of AIDS-associated Cryptococcus neoformans meningitis (CM) in Kigali, Rwanda, during 1983-1992, and to highlight some diagnostic and epidemiological features of the disease. METHODS: during the study period, 3476 cerebrospinal fluid (CSF) specimens from 2824 adults (1578 men, 1246 women) were analysed in the Laboratory of Microbiology at the Centre Hospitalier de Kigali, Rwanda, Central Africa, using direct examination, culture and detection of the cryptococcal antigen (CrAg) in the CSF. RESULTS: CM was diagnosed among 549 (19%) patients (347 men, 202 women) and was by far the leading cause of meningitis before Neisseria meningitidis (n=115), Streptococcus pneumoniae (n=68), Mycobacterium tuberculosis (n=26). E. coli, Klebsiella pneumoniae, non-typhoid Salmonella (n=l5) and streptococci (n=4). The number of CM increased from one case in 1983 to 130 new cases in 1992. All 293 tested CM patients had HIV-1 antibodies. The male/female ratio declined from 3.31 during 1983-1987 to 1.58 during 1988-1992. CM showed a seasonal fluctuation, the highest number of infections being observed during the long rainy season. The sensitivity and specificity of the latex test for diagnosing CM was 98% and 99%, respectively. Cryptococcus neoformans var. gattii was cultured from eight (1.6%) of the 499 culture positive patients. CONCLUSION: CM is an important opportunistic infection among AIDS patients in Central Africa. It remains a problematic diagnosis in areas with limited diagnostic facilities. PMID- 10468127 TI - Immune responses to administration of a vaccine against Haemophilus influenzae type B in splenectomized and non-splenectomized patients. AB - OBJECTIVES: we investigated the cause of hypo-responsiveness to vaccines in splenectomized subjects. METHODS: we evaluated the immune responses to a Haemophilus influenzae type b vaccine and the sizes of lymphocyte subpopulations in 25 splenectomized and 45 non-splenectomized thalassaemic patients, in 12 individuals who had been splenectomized after trauma and in 20 controls. RESULTS: the immune response in the controls was significantly higher (P < 0.001) than in splenectomized patients after trauma and in both, the response was higher (P < 0.001) than in thalassaemic patients. In asplenic subjects after trauma, percentages of CD3 and CD4 cells were lower (P < 0.001) than in patients in the other groups; the controls had higher percentages of CD8 cells (P < 0.001) than patients in the other groups. The natural logarithm of the mean percentage of (CD19 showed a quadratic trend from thalassaemic patients through asplenic subjects to controls (P < 0.001). Levels of CD16+ natural killer (NK) cells were higher (P < 0.001) only in asplenic subjects after trauma. CONCLUSIONS: the significant decrease in the immune response of the splenectomized thalassaemic patients vs. non-splenectomized thalassaemic patients may, in part, be due to their basic immunological condition. Thus, the best strategy for protecting these subjects is to vaccinate them before the splenectomy. PMID- 10468128 TI - Meningococcal carriage in relation to an outbreak of invasive disease due to Neisseria meningitidis serogroup C in the Netherlands. AB - BACKGROUND: a cross-sectional study on meningococcal carriage was performed in Putten, a small rural town in the Netherlands where an unusual high incidence of invasive meningococcal disease (IMD) due to Neisseria meningitidis C:2a:P1.5 occurred. The outbreak was controlled by mass vaccination of all inhabitants aged 2 to 20 years. METHODS: meningococcal carriage was studied in three groups: (1) a systematic age-specific sample of 2-20 year olds who visited the immunization clinic in Putten (January 1998: n=411); (2) children and adolescents in the same age range recruited through a kindergarten and schools in Venlo, a town where the causative strain of IMD had not been encountered (February 1998; n=374); (3) all initial carriers in Putten and a sample of non-carriers in that town (March 1998: n=145). Oropharyngeal swabs were taken for the purpose of isolating N. menigitidis. RESULTS: the prevalence of carriage was 12.4% in Putten and 18.2%, in Venlo, but the prevalence of group C meningococci was higher in Putten (1.7%) than Venlo (0.5%). N. meningitidis C:2a:P1.5 was isolated twice in Putten and not at all in Venlo. A second examination in Putten showed that 18 of the 22 repeatedly tested carriers were still carriers, and six new carriers were found among the 55 initial non-carriers. Of the two known carriers of C:2a:P1.5, one was still carrying the same strain, and the other did not participate in the second investigation. Carriage was associated with increasing family size, discotheque visits and visits to youth clubs and sports clubs. In contrast, visits to the swimming pool appeared to be related to a lower risk, as was recent antibiotic use. CONCLUSION: the prevalence of carriage with the invasive strain C:2a:P1.5 was low in the population that experienced a community-wide outbreak recently: the specific strain was not found in the reference population. This indicates a relatively high risk of developing the invasive disease for those who become infected with such strains. PMID- 10468129 TI - Maternal infectious morbidity following multiple courses of betamethasone. AB - OBJECTIVE: the beneficial effects of antepartum corticosteroids on the reduction of morbidity and mortality in the premature neonate have been amply demonstrated. The NIH consensus statement has, therefore, endorsed their use in women at risk from pre-term delivery between 24 and 34 weeks gestation. Patients at persistent risk of pre-term delivery nay receive multiple weekly courses. However, increased susceptibility to infection is a well-recognized complication of prolonged high dose steroid therapy. We examined infectious morbidity among women exposed to three or more courses of betamethasone. METHODS AND OUTCOMES MEASURES: thirty seven patients at risk of pre-term delivery who received three or more courses of betamethasone (median = 6: range 3-10) and 74 normal controls, matched for maternal age, route of delivery, and year of delivery were included in the study. Data on medical care provided to study and control patients between 24 weeks gestation and 6 weeks postpartum were retrieved from centralized medical records. Incidences and types of infections were compared by Chi-square and Fisher's exact test, as appropriate. Only infections diagnosed at least 1 week after betamethasone therapy was initiated were included. Patients with pre-existing conditions predisposing to infectious morbidity were excluded. RESULTS: twenty four of 37 patients (64.8%) exposed to betamethasone vs. 13 of 74 (17.5%) controls developed infectious diseases (P < 0.001). Symptomatic lower urinary tract infections occurred in 13 of 37 (35.1%) and two of 74 (2.7%) in the study and control groups, respectively (P < 0.001). Serious bacterial infections were found in nine of 37 (24.3%) vs. none of 74 (0%) patients, respectively (P < 0.001). These included sepsis (n=2), pneumonia (n=4), pyelonephritis (n=2), and cholangitis (n=1). Eight of nine serious infections occurred in patients exposed to five or more weekly courses of betamethasone. Postpartum endometritis related to Caesarean delivery was found in five of 37 patients (13.5%) vs. seven of 74 (9.4%), respectively (P=NS). CONCLUSIONS: Three or more courses of antepartum betamethasone in women at risk of pre-term delivery are associated with substantial infectious maternal morbidity. The excess morbidity consisted mainly of bacterial infections, some of which were associated with systemic and potentially life-threatening infections. PMID- 10468130 TI - Cerebrospinal fluid cytokine levels and dexamethasone therapy in bacterial meningitis. AB - OBJECTIVES: cerebrospinal fluid (CSF) levels of interleukin (IL)-1 beta and tumor necrosis factor (TNF) alpha were measured to assess the effect and application of dexamethasone (Dex) therapy for bacterial meningitis. METHODS: associations between clinical findings and CSF parameters were first investigated, and prognosis was compared between 25 patients with Dex and 12 without Dex therapy. RESULTS: patients with the presence of disturbed consciousness showed higher CSF levels of TNF alpha (mean: 3015 pg/ml) or protein (mean: 215 mg/dl) than those without it (both, P < 0.O5). Simultaneous increase of TNF alpha (> 1000 pg/ml) and protein (> 100 g/dl) was observed in 80%, of patients with profoundly disturbed consciousness. Patients with Dex therapy presented higher TNF alpha/protein levels at diagnosis than those without Dex therapy (P < 0.05). Despite worse conditions at diagnosis, only one of 14 Dex-treated patients whose initial CSF TNF alpha levels exceeded 1000 pg/ml developed deafness. On the other hand, two of four patients without Dex therapy who had the same TNF alpha level suffered from psychomotor retardation. The differences in the frequency of sequelae between those with and without Dex therapy were significant in patients showing high TNF alpha level (P < O.05), but not in those showing high CSF levels of IL-1 beta or protein. The logistic regression analysis indicated that high CSF protein level (P < O.0001), or no Dex therapy (P=0.0001) was the independent risk factor for sequelae. CONCLUSIONS: although the study number was small, our observations suggested that CSF TNF alpha/protein levels reflected the neurologic severity, and implied that early Dex therapy might be beneficial for patients with prominently high TNF alpha levels. PMID- 10468131 TI - High rates of disseminated infection due to non-tuberculous mycobacteria among AIDS patients in Finland. AB - OBJECTIVE: to determine the rate of disseminated infection due to non-tuberculous mycobacteria (NTM) among Finnish AIDS patients, and to analyse the epidemiology of these infections. METHODS: in a prospective cohort study HIV-infected patients with CD4 counts < 200 x 10(6)/l were interviewed, and had mycobacterial blood cultures performed at baseline and at 6 months, then subsequently for clinical indications; autopsies were performed on patients who died. The cohort was followed at least for 24 months or to death. Water samples were collected from the homes of patients and from the environment and cultured for organisms of the Myobacterium avium complex (MAC). Environmental and clinical isolates were compared using pulsed field gel electrophoresis (PFGE). RESULTS: NTM infection occurred in 22 (43%) of 51, 19 isolates were Mycobacterium avium, two M. genavense and one M. intracellulare. Multivariate analysis identified urban residence (P=0.04) and eating raw fish (P=0.04) as independent risk factors. Molecular analysis revealed two clusters of related isolates (three M. avium, two M. genavense) among urban residents. CONCLUSION: AIDS patients in Finland have high rates of disseminated infection due to NTM. Clusters of identical organisms and association with urban residence suggests that these are newly acquired infections in advanced AIDS. PMID- 10468132 TI - Parvovirus B19 infection induces apoptosis of erythroid cells in vitro and in vivo. AB - OBJECTIVE: intrauterine parvovirus B19 infection is related to non-immune hydrops fetalis, the pathogenesis of which is based on the strict tropism of B19 for erythroid precursor cells and the massive destruction of the infected erythroid cells, although the mechanism of beta19-induced cytotoxicity has not been studied in detail. The purpose of this study is to provide empirical evidence that beta19 induces apoptosis of erythroid cells both in vitro and ill vivo. METHODS: we analysed culture cells infected in vitro by B19 and tissues of nine cases of hydrops fetalis caused by B19 intrauterine infection by histological and biological methods. RESULTS: cells infected iil vitro by B19 showed nuclear changes characteristic of apoptosis by light microscopic examination and DNA extracted from the infected cells was fragmented. Electron microscopic examination showed the nuclei of infected cells contained crescent-shaped clumps of heterochromatin with increased density and double staining with anti-B1 9 antibody and terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate-digoxigenin nick-end labeling (TUNEL) confirmed apoptosis of individual cells. Tissues of cases of hydrops fetalis caused by B19 contained erythroid cells with nuclear inclusions and characteristic nuclear changes of apoptosis by light microscopy. The double-staining confirmed apoptosis of erythroid cells in the tissues. Immunohistochemical analysis with antibodies against cellular factors involved in apoptosis showed that caspase3, p53 and p21 were positive in infected cells. PMID- 10468133 TI - Diagnosis of Pneumocystis carinii pneumonia: immunofluorescence staining, simple PCR or nPCR. AB - OBJECTIVES: to compare immunofluorescence (IF) test, routinely used in the department for the detection of Pnemocystis carinni with simple polymerase chain reaction (PCR) and nested PCR (nPCR) METHODS: Bronchoalveolar lavage (BAL) and induced sputum (IS) specimens from HIV-positive (39), lung transplant ssart transplant (2), and one each from non-Hodgkin's lymphoma, drug addict and a premature baby were screened by IF test, simple PCR and nPCR for the presence of P.carinii. RESULTS: of the 46 specimens tested, two (4.3%) were positive by IF, 11 (23.9%) by simple PCR and 21 (45.6%) by nPCR. Both simple and nPCR amplified those found positive by IF test. Analysis of the clinical data revealed both IF positive, 10 of the simple PCR and 15 of the nPCR group were strongly suspected of P. carinii pneumonia (PCP). Two specimens, one from a patient where chest X ray was suggestive of PCP and the other where post-mortem histology revealed the presence of PCP, were negative by IF test. CONCLUSION: simple PCR detection may be considered for patients where PCP is suggestive clinically and the specimen is negative by IF test. PMID- 10468134 TI - Anti-neutrophil cytoplasmic autoantibodies (ANCA) to bactericidal/permeability increasing (BPI) protein recognize the carboxyl terminal domain. AB - OBJECTIVES: to identify the region of bactericidal/permeability-increasing protein (BPI) recognized by anti-BPI ANCA. METHODS: sera from 140 patients with a variety of clinical diagnoses (20 systemic vasculitis, 12 cystic fibrosis, 22 bronchiectasis/chronic obstructive airways disease, three diabetes mellitus, 13 chronic renal failure, 12 primary sclerosing cholangitis, eight ulcerative colitis, three Crohn's disease, seven cancer, and 40 other or unknown diagnoses) known to be reactive against native (nBPI), were screened by solid phase enzyme linked immunosorbent assay (ELISA) against a panel of recombinant fusion proteins; holo BPI (rBPI), recombinant lipopolysaccharide binding protein (rLBP), an N-terminal fragment of rBPI (rBPI21 ) and 'fusion' proteins containing the C- or N-terminal ends of BPI spliced with N-or C-ends of LBP, respectively. RESULTS: a strong correlation was seen between the degree of reactivity to rBPI and the BPI C-terminal fusion protein, r=0.69, P < 0.001, as well as between nBPI and rBPI protein, r=0.55, P < 0.001, but not between nBPI and the N-terminal region of BPI (rBPI21), or proteins containing only the N-terminal fragment. Binding to proteins containing the BPI C-terminus was confirmed to be specific by fluid phase inhibition ELISA and Western blot analyses. CONCLUSIONS: together these data suggest that circulating autoantibodies to BPI from patients with different diseases recognize the C-terminal region of BPI. PMID- 10468135 TI - Acute disseminated histoplasmosis complicated with hypercalcaemia. AB - A case of acute progressive disseminated histoplasmosis complicated with hypercalcemia is reported and the literature is reviewed. This and the previously reported cases imply that physicians should have a higher index of suspicion for this infection and the probable underlying diseases resulting from impaired cellular-mediated immunity when encountering patients with hypercalcaemia. PMID- 10468136 TI - Mycoplasma hominis bacteraemia not associated with genital infections. AB - We describe two cases of systemic infection with M. hominis without prior genital infection and review the previous literature on this condition. PMID- 10468137 TI - Clostridium difficile infection of a prosthetic joint presenting 12 months after antibiotic-associated diarrhoea. AB - Clostridium difficile is a common cause of antibiotic-associated diarrhoea, but it rarely causes extra-colonic disease. An 83-year-old woman who developed culture-positive C. difficile-associated diarrhoea following pneumonia and a total hip replacement was treated successfully with oral metronidazole therapy. She was readmitted 12 months later for revision of the hip prosthesis because of increasing pain. At surgery an abscess associated with the prosthesis was discovered. Culture of the pus grew C. difficile. When the surgical isolate and the cryopreserved faecal isolates were compared using Pulsed Field Gel Electrophoresis (PFGE), an identical banding pattern was observed. This case extends the clinical spectrum of C. difficile infection by documenting a prolonged interval between the initial infection, and diagnosis of the prosthetic joint infection. The importance of molecular typing to show strain relatedness is demonstrated. PMID- 10468138 TI - Shiga toxin 2 promotes the stem cell differentiation into granulocytes in the bone marrow causing marked granulocytosis in the peripheral blood. PMID- 10468139 TI - An unusual case of mixed bacterial meningitis in an immunocompetent adult. PMID- 10468140 TI - Recent clinical trials in the diagnosis of deep-vein thrombosis. AB - Contrast venography remains the gold standard for the diagnosis of deep-vein thrombosis (DVT). However, an ongoing interest exists in developing safe diagnostic strategies that are less invasive, less expensive, widely and readily available, and more acceptable to patients than venography. For patients with symptoms suggestive of DVT, ultrasound is now the most frequently used test. Recent clinical outcome studies have evaluated management strategies that use ultrasound testing, either alone or with adjunct D-dimer testing or clinical risk stratification. Based on current evidence, a noninvasive strategy using compression ultrasound has been shown to be a safe alternative for patients suspected of having DVT, and an approach using D-dimer also appears promising. Neither ultrasound nor other noninvasive tests are adequately sensitive when used with patients who do not have leg symptoms but who are at high risk for DVT (eg, recent total knee or hip arthroplasty). Thus, no currently available noninvasive testing strategy can reliably exclude DVT in asymptomatic patients. PMID- 10468141 TI - Diagnosis of pulmonary embolism. AB - The patient with suspected pulmonary embolism presents a challenging diagnostic problem. The symptoms and signs are nonspecific, and objective testing is required to establish or exclude the presence of pulmonary embolism. Lung scanning continues to be a first-line test, but in 40% to 70% of all patients, the results do not definitively provide indication for either giving or withholding anticoagulant treatment even when combined with the clinical assessment. Pulmonary angiography is the reference standard, but it is invasive and may not be available in all clinical settings. Pulmonary embolism is strongly associated with proximal deep-vein thrombosis of the legs (popliteal, femoral, or iliac vein thrombosis). Objective testing for proximal deep-vein thrombosis is useful in patients with suspected pulmonary embolism. A positive result from such testing provides an indication for anticoagulant treatment. Serial testing for proximal deep-vein thrombosis is a safe and effective alternative to pulmonary angiography in patients with adequate cardiorespiratory reserve. The assay for plasma D-dimer using either a rapid enzyme-linked immunospecific assay technique or a bedside whole-blood agglutination technique is promising as a test for excluding venous thromboembolism. A positive result by spiral CT imaging is useful for ruling in a diagnosis of pulmonary embolism, but the safety of withholding treatment in patients with negative spiral CT results remains uncertain. Pulmonary angiography continues to have an important role in selected patients in whom it is critical to definitively confirm or exclude the presence of pulmonary embolism. PMID- 10468142 TI - Prevention of venous thromboembolism. AB - Venous thromboembolism is a preventable cause of morbidity and death in hospitalized patients. In addition to administering correct and effective prophylaxis to the various treatment groups, therapy should be tailored to the individual based on the identification of baseline risk factors. No patient at significant risk for venous thrombosis should be left unprotected. PMID- 10468143 TI - Thrombophilia. AB - Thrombophilia is now considered a multicausal disease, with an interplay of acquired and genetic risk factors. Recent studies have shown that patients with the 20210 A prothrombin mutation display remarkably similar characteristics compared with patients with Factor V Leiden mutation. It is evident that neither the Factor V Leiden mutation nor the 20210 A prothrombin mutation is a major risk factor for myocardial infarction or stroke, unless accompanied by other classical risk factors, including diabetes mellitus, hypertension and smoking. Finally, the homozygous form of the thermolabile methylenetetrahydrofolate reductase gene, although leading to elevated homocysteine levels, seems not to represent a genetic risk factor for venous thrombosis. PMID- 10468144 TI - Thrombin inhibitors as anticoagulant agents. AB - Hirudin and its analogues and the synthetic antithrombin agents are interesting new antithrombotic agents that have been studied in a number of well-designed randomized clinical trials and further studies are underway. These agents offer certain advantages over heparin and low-molecular-weight heparin, and at least one agent is orally bioavailable. Studies have shown that the specific thrombin inhibitors can significantly decrease the incidence of composite cardiac endpoints in acute ischemic syndromes (following thrombolysis for myocardial infarction, unstable angina, and non-Q wave myocardial infarction and coronary angioplasty), but it is disappointing that the benefits obtained during short term treatment are not sustained in the long term. Recent data are reviewed here from clinical trials supporting the use of the specific antithrombin agents in the treatment of acute cardiac ischemic syndromes, the prevention and treatment of venous thromboembolism, and the management of heparin-induced thrombocytopenia. PMID- 10468145 TI - Thrombolytic therapy for venous thromboembolism. AB - The role of thrombolytic therapy for venous thromboembolism remains uncertain. To date, no well-designed randomized clinical trial has shown that the benefits of thrombolysis exceed the risks for a well-defined subgroup of patients with acute pulmonary embolism. Recent reports have underscored the risks of thrombolytic therapy. The largest multinational registry of patients in whom acute pulmonary embolism had been diagnosed found that 3% of 311 patients who underwent thrombolysis for acute pulmonary embolism suffered an intracranial hemorrhage. Because of the serious risk associated with systemic thrombolysis, investigators have continued to study methods that rapidly remove venous thromboemboli while minimizing the risk. Reports of catheter fragmentation combined with thrombolysis as well as catheter-directed thrombolysis are promising, but well-designed clinical trials are needed to clarify the utility of these techniques. PMID- 10468146 TI - Thrombolytic therapy for arterial thrombosis. AB - The major development in the field of intra-arterial thrombolytic therapy over the past year was the publication of the phase II results of the Thrombolysis or Peripheral Arterial Surgery study, which compared the safety and efficacy of catheter-directed thrombolysis and surgery as the initial treatment of acute arterial occlusion. The results are consistent with those of the prior two studies, showing little or no difference between surgery and thrombolysis in the most important endpoints of survival and amputation rate. Patients receiving thrombolysis needed fewer interventions, but this benefit was balanced by increased bleeding complications. Additional studies have, therefore, been aimed at identifying subsets of patients with acute arterial occlusion who are most likely to benefit from thrombolysis. These studies have refined the selection criteria for use of thrombolytic therapy over the past year. In addition, studies have been published evaluating new drug doses and regimens aimed at broadening the scope of thrombolytic therapy in patients with acute arterial occlusion. PMID- 10468147 TI - Autoimmune factor VIII inhibitors. AB - Dramatic hemorrhage can follow the rare, spontaneous development of inhibitory autoantibodies to factor VIII (FVIII). Diagnosis, which is often delayed, relies on complex, interpretive testing for presence and titer of the inhibitor antibody. Low cross reactivity of the inhibitor to porcine FVIII supports consideration of its therapeutic use. Recombinant activated factor VII has expanded available therapeutic options beyond prothrombin complex concentrates and their activated forms. Use of genetically engineered FVIII molecules has further defined immunodominant epitopes on FVIII and may provide a therapeutic alternative. The optimal region of immunosuppressive therapy remains to be defined. Future laboratory and clinical studies are necessary for advancement of pathophysiologic knowledge and therapeutic options for patients with this uncommon but clinically important disorder. PMID- 10468148 TI - Acquired autoantibodies to coagulation factors. AB - Acquired coagulation factor antibodies are either alloantibodies or autoantibodies. Alloantibodies are formed when the body reacts to an external antigen. Commonly, alloantibodies arise when a patient who has a congenital clotting factor deficiency is infused with a blood product. Alternately, patients exposed to a coagulation protein from a different species may develop alloantibodies to the animal protein that cross-reacts with their own protein. On the other hand, autoantibodies develop spontaneously in people without pre existing factor deficiencies and without exposure to an external antigen. These antibodies may neutralize function of a clotting factor, promote rapid clearance of a clotting factor from the blood, or alter the clotting factor in such a way that the protein-antibody complex acquires a unique function. This review focuses on recent reports of autoantibodies directed against fibrinogen, prothrombin, factor V, factor VII, factor X, and von Willebrand factor, in which these various activities of autoantibodies are illustrated. PMID- 10468149 TI - Management of bleeding with uremia and liver disease. AB - Bleeding can complicate the clinical course of both liver disease and uremia. The pathogenesis of bleeding in both syndromes is complex. A variety of options are available for treatment of uremic bleeding including desmopressin, erythropoietin, and estrogens. Bleeding complications of liver disease reflect the importance of this organ for the production of both coagulation factors and thrombopoietin. Careful assessment of both the patient's clinical situation and laboratory findings is important for tailoring therapy of the bleeding patient. PMID- 10468150 TI - Bleeding complications with platelet glycoprotein IIb/IIIa receptor antagonists. AB - Platelet glycoprotein (GP) IIb/IIIa receptor antagonists are being used with increasing frequency in the settings of percutaneous coronary interventions and acute ischemic syndromes. The development of bleeding complications following GPIIb/IIIa blockade represents a significant limitation to its effectiveness. Baseline characteristics predictive of future bleeding events in patients receiving platelet GPIIb/IIIa receptor antagonist include older age, low body weight, evolving myocardial infarction, and female sex. In patients undergoing percutaneous coronary interventions with adjunctive GPIIb/IIIa inhibition, the risk of bleeding, particularly from the femoral vascular access site, may be reduced through the use of low-dose, weight-adjusted heparin (70 U/kg), avoidance of postprocedural heparin, and early vascular sheath removal. Strategies to reduce the incidence of bleeding complications in patients receiving GPIIb/IIIa inhibitors are proposed in this article. PMID- 10468151 TI - Laboratory markers of platelet activation and their clinical significance. AB - Whole blood flow cytometry is a powerful new laboratory technique for assessment of platelet activation and function. Flow cytometry can be used to measure platelet hyperreactivity, circulating activated platelets, leukocyte-platelet aggregates, and procoagulant platelet-derived microparticles in a number of clinical settings, including acute coronary syndromes, angioplasty, cardiopulmonary bypass, acute cerebrovascular ischemia, peripheral vascular disease, diabetes mellitus, preeclampsia, and Alzheimer's disease. Clinical applications of whole blood flow cytometric assays of platelet function in these diseases may include identification of patients who would benefit from additional antiplatelet therapy and prediction of ischemic events. Circulating monocyte platelet aggregates appear to be a more sensitive marker of in vivo platelet activation than circulating P-selectin-positive platelets. Flow cytometry can also be used in the following clinical settings: monitoring of glycoprotein IIb IIIa antagonist therapy, diagnosis of inherited deficiencies of platelet surface glycoproteins, diagnosis of storage pool disease, diagnosis of heparin-induced thrombocytopenia, and measurement of the rate of thrombopoiesis. PMID- 10468152 TI - Drug-induced thrombocytopenia. AB - Many drugs can induce thrombocytopenia mediated by drug-dependent antiplatelet antibodies. Recent studies have documented specific epitopes for drug-dependent antibody binding on glycoprotein Ib-IX, glycoprotein IIb-IIIa, and platelet endothelial cell adhesion molecule-1. Molecular identification of antibody binding sites may help to identify susceptible individuals. Management of patients with unexpected thrombocytopenia who are taking multiple drugs remains a difficult clinical problem. A recent systematic review of all published case reports of drug-induced thrombocytopenia ranks drugs according to the strength of clinical evidence for a causal relation to thrombocytopenia. This database is available online at http://moon.ouhsc.edu/jgeorge. PMID- 10468153 TI - Megakaryocyte structure and function. AB - Recent advances in the understanding of megakaryocyte (MK) function largely have been made through the careful observation of the morphological and structural events underlying MK development. Ultrastructural localization of enzymatic activities has facilitated the specific recognition of their committed diploid precursors. Observation of the sequential features of endomitosis demonstrates that although similar to normal mitosis, cell division aborts at the anaphase stage. The ability of thrombopoietin to induce the full maturation MKs in vitro not only facilitates platelet release but has increased our knowledge of various subcellular aspects of the phenomenon and eventually will improve the in vivo detection of the site of platelet formation and shedding. Finally, the structural and functional consequences of MK molecular dysfunction leading to thrombocytopenia or myelofibrosis can now be investigated because of the development of transgenic animal models. This review aims to incorporate these new findings within the classical knowledge of MK structure related to its function. PMID- 10468154 TI - Bibliography. Current world literature. Hemostasis and thrombosis. PMID- 10468155 TI - Zinc protects against ultraviolet A1-induced DNA damage and apoptosis in cultured human fibroblasts. AB - Ultraviolet A1 (UVA1) radiation generates reactive oxygen species and the oxidative stress is known as a mediator of DNA damage and of apoptosis. We exposed cultured human cutaneous fibroblasts to UVA1 radiation (wavelengths in the 340-450-nm range with emission peak at 365 nm) and, using the alkaline unwinding method, we showed an immediate significant increase of DNA strand breaks in exposed cells. Apoptosis was determined by detecting cytoplasmic nucleosomes (enzyme-linked immunosorbent assay method) at different time points in fibroblasts exposed to different irradiation doses. In our conditions, UVA1 radiation induced an early (8 h) and a delayed (18 h) apoptosis. Delayed apoptosis increased in a UVA dose-dependent manner. Zinc is an important metal for DNA protection and has been shown to have inhibitory effects on apoptosis. The addition of zinc (6.5 mg/L) as zinc chloride to the culture medium significantly decreased immediate DNA strand breaks in human skin fibroblasts. Moreover, zinc chloride significantly decreased UVA1-induced early and delayed apoptosis. Thus, these data show for the first time in normal cutaneous cultured cells that UVA1 radiation induces apoptosis. This apoptosis is biphasic and appears higher 18 h after the stress. Zinc supplementation can prevent both immediate DNA strand breakage and early and delayed apoptosis, suggesting that this metal could be of interest for skin cell protection against UVA1 irradiation. PMID- 10468156 TI - Ferrous sulfate toxicity: a review of autopsy findings. AB - Ferrous sulfate is the leading cause of accidental pediatric poisonings. Despite the requirement for child-resistant packaging for any oral iron product with 250 mg or more per container, the incidence has continued to increase. Although the clinical presentation of iron toxicity has been well described, pathologic findings in human tissue and correlation with clinical data are scant. We reviewed autopsies from the Armed Forces Institute of Pathology of 11 children who died from ferrous sulfate toxicity. Clinical data, morphologic changes, and iron levels in tissue were evaluated. The children's ages ranged from 11 to 36 mo. Prominent iron deposition in gastric and small intestinal mucosa was associated with necrosis, with some cases demonstrating prominent vascular iron deposition. The clinical courses were rapid and progressed from Stage I to Stage III. These observations were correlated with increased levels of iron in various tissues, as determined by analytical atomic absorption spectrophotometry. The morphologic and chemical analysis data provide information on the pathogenesis of ferrous sulfate poisoning; the vascular iron deposition may be related to subsequent hemorrhage. In the liver the periportal necrosis is probably a direct cytopathic effect of the highest levels of iron carried to these cells by the portal blood flow. PMID- 10468157 TI - Protective role of intraperitoneally administered vitamin E and selenium in rats anesthetized with enflurane. AB - The aim of this investigation was to determine levels of liver vitamins A and E and blood biochemical and hematological parameters in the enflurane anesthesia of rats. Fifty adult male Wistar rats were used in this study. All rats were randomly divided into five groups. The first and second groups were used as the control and anesthesia control groups, respectively, and only the placebo was intraperitoneally injected. The third group was intraperitoneally administered with vitamin E (dl-alpha-tocopheryl acetate, 100 mg/kg body weight), the fourth group with Se (Na2SeO3 1.5 mg/kg body weight), and the fifth group with vitamin E and Se (dl-alpha-tocopheryl acetate, 100 mg/kg body weight + Na2SeO3 1.5 mg/kg body weight). This administration was done for three times with overday intervals and the second, third, forth, and fifth group rats were taken to enflurane anesthetise for 2 h. The liver vitamin E level was slightly lower in the anesthesia control group than in control group. However, the liver vitamin E content was significantly (p < 0.05 and p < 0.01) increased in vitamin E, Se, and combination groups, whereas the vitamin A level in liver was not statistically different. In general, plasma levels of alanine aminotransferase, creatin kinase, total bilirubin, urea, red blood cell counts, packet cell volume, and hemoglobulin values were significantly (p < 0.05 and p < 0.001) increased during the anesthesia and returned to near control values after the vitamin E plus selenium injection. However, administration of vitamin E had less effect on the hematological and biochemical parameters compared to that of selenium and their combination with vitamin E. However, the white blood cell count and levels of alkaline phosphatase, aspartate aminotransferase, total cholesterol, triglycerides, total protein, and creatinine were not statistically influenced by the anesthesia. In conclusion, we observed that plasma levels of some enzymes and metabolites were significantly increased in the enflurane anesthesia of rats, whereas the liver vitamin E levels were slightly decreased. Therefore, we observed that vitamin E and selenium have a protective effect against anesthesia complication, but the effect of selenium appears to be much greater than the vitamin E. PMID- 10468158 TI - Iodine and zinc, but not selenium and copper, deficiency exists in a male Turkish population with endemic goiter. AB - Although endemic goiter has been shown to have a high prevalence in Turkey, little is known about the concentration of urinary iodine, plasma selenium (Se), copper (Cu), and zinc (Zn) in these patients. We studied on 140 male patient with endemic goiter (mean age: 22.2 +/- 0.19 yr) and 140 healthy male subjects (mean age: 21.8 +/- 0.28 yr). Daily urinary iodine excretion was determined by the ionometric method. Plasma Se, Zn, and Cu were determined by using atomic absorption spectrometry. Daily urinary iodine excretion was found to be significantly lower in the patient group (38.7 +/- 2.26 microg/d) than that of controls (50.73 +/- 2.56 microg/day, p = 0.001). Plasma Zn concentrations were also found to be significantly lower in the patient group (1.04 +/- 0.03 microg/mL) than that of controls (1.16 +/- 0.02 microg/mL, p = 0.001). No significant difference was determined in Se and Cu concentrations between the patient and control groups. Our study shows that a moderate iodine deficiency exists in both patients with endemic goiter and control subjects, which indicates the important role of iodine deficiency in the etiopathogenesis of endemic goiter in Turkey. Zinc deficiency may also contribute to the pathogenesis of endemic goiter. However, Se and Cu do not seem to have any role in the etiopathogenesis of endemic goiter in Turkey. A community-based iodine fortification program throughout the country may be proposed to take over the problem, which also can prevent the contributing effects of other element deficiencies that occur when iodine deficiency is the prevailing factor. PMID- 10468159 TI - Accelerated volatilization rates of selenium from different soils. AB - Selenium (Se), an element found naturally in a variety of soils, can accumulate in drainage water of lands under intensive irrigation, even reaching levels that are toxic to mammals and birds. Volatilization of Se by soil microorganisms into dimethylselenide (DMSe) can be enhanced by certain soil amendments and, thus, be used as a soil remediation process. In an 8-wk laboratory study, five soils from California and one from Germany were spiked with 75SeO3(2-) (22.3 mg/kg Se). Two amino acids (DL-homocysteine and L-methionine), a carbohydrate (pectin), and a protein (zein) were tested as soil amendments. Gaseous 75Se emissions were trapped with activated carbon and measured in a gamma counter. Depending on soil type, the cumulative volatilization from the control flasks varied between 1.2% and 9.0% of applied 75Se. Both zein and L-methionine strongly increased volatilization (max. 43% of 75Se applied), whereas DL-homocysteine had a much smaller stimulating effect. Pectin showed a moderate effect, but enhanced Se volatilization rates were sustained much longer when compared to the zein amendment. Volatilization rates of Se followed a simple first-order reaction. Gaseous Se emission in the soils treated with L-methionine yielded an S-shaped curve, which fit a growth-modified first-order rate model. Although zein and L methionine were the most favorable treatments enhancing Se volatilization, all six soils responded differently to the soil amendments. PMID- 10468160 TI - Element content of human umbilical artery and vein in umbilical cord. AB - To elucidate the element content of newborn blood vessels, umbilical arteries and veins in human umbilical cords, which had the advantage of easy sampling, were examined by ICP-AES. Umbilical cords were removed after birth. Mothers' ages ranged from 26 to 35 yr. It was found that the content of sulfur was the highest in both umbilical arteries and veins, being higher than the content of calcium and phosphorus. With respect of the content of sulfur, calcium, and magnesium, there were significant differences between the arteries and veins. PMID- 10468161 TI - A possible balance of phosphorus accumulations among bone, cartilage, artery, and vein in single human individuals. AB - To elucidate relationships between the decrease of mineral contents in human bones and the accumulation of minerals in the other human tissues, the contents of phosphorus in human bones, arteries, veins, and cartilages in 27 subjects (17 men and 10 women) were analyzed by inductively coupled plasma-atomic emission spectrometry. These were resected from subjects who died in the age range 40-98 yr. Calcanei were chosen for analysis of mineral contents in contrast to arteries such as the femoral, popliteal, and common carotid arteries, veins such as superior and inferior venae cavae, internal jugular, and femoral veins, and pubic symphyses. It was found that the content of phosphorus in calcanei was in agreement with that in both the pubic symphysis and the arteries such as femoral, popliteal, and common carotid arteries, but it was not in agreement with that in the veins such as superior and inferior venae cavae, internal jugular, and femoral veins. This suggests that phosphorus released from bones is accompanied by accumulations of phosphorus in the artery and cartilage. PMID- 10468162 TI - Prevention by sodium 4,5-dihydroxybenzene-1,3-disulfonate (Tiron) of vanadium induced behavioral toxicity in rats. AB - Recent studies have shown that oral vanadate (V5+) administration results in behavioral toxicity in rats. The chelating agent Tiron (sodium 4,5 dihydroxybenzene-1,3-disulfonate) is an effective antidote in the removal of vanadium from vanadium-loaded rats. In this study, the protective activity of Tiron on vanadate-induced behavioral toxicity was evaluated in adult rats. Intraperitoneal treatment with Tiron at 235 or 470 mg/kg was initiated after 6 wk of oral sodium metavanadate administration (16 mg/kg/d) and continued for 2 wk. Although vanadate exposure did not result in a significant reduction in the general activity of the animals in an open field, a lower active avoidance acquisition could be observed. However, the vanadate-induced behavioral deficit was reverted by Tiron administration at 470 mg/kg. The present results suggest that Tiron may protect, at least in part, against metavanadate-induced behavioral toxicity. PMID- 10468163 TI - Zinc coadministration attenuates melatonin's effect on nitric oxide production in mice. AB - Both melatonin (MEL) and zinc (Zn) are considered beneficial for anti immunosenescence. MEL's effects on immune functions are partly attributed to an interaction with Zn. However, the augmentation of or interference with MEL's effects by coadministration of Zn remains unclear. In this study, adult older mice received either MEL (10 microg/mL), Zn (22 microg/mL), MEL+Zn, or null supplementation from drinking water for 3 mo. The results showed that treated mice, irrespective of the type of added chemicals, had higher body-weight gain and body-fat content than control mice. MEL- and Zn-treated mice also had increased serum free fatty acid levels. In addition, the MEL group had decreased serum NOx (nitrite+nitrate) values. Serum tumor necrosis factor-alpha levels were increased, although nonsignificantly, in mice that received either MEL or Zn supplementation. However, the differences described were not retained in the mice that received MEL+Zn treatment. We conclude that a high-dose Zn coadministration might exert negative influences on MEL's regulatory effects, at least on nitric oxide production. PMID- 10468164 TI - Hair concentrations of calcium, iron, and zinc in pregnant women and effects of supplementation. AB - In this investigation, the concentration levels of hair elements of calcium, iron, and zinc were measured in pregnant women from Tianjin metropolis, China. The subjects were 93 cases of pregnant women who had been suffering from calcium, iron, or zinc deficiency judged by blood tests at the mid-term of the second trimester or early in the third trimester. Of these 93 cases, 82 subjects had their hair element levels measured when the blood tests were conducted. Then, they were supplied with mineral element nutrients of gluconic acidic zinc (noted as Zn-nutrient), gluconic acidic calcium (Ca-nutrient), or/and ferrous sulfate (Fe-nutrient) which were correspondent to the deficient element(s) for more than 2 mo before 84 subjects returned to hospital for further diagnoses and had their hair element levels measured for the second time. Finally, in the third trimester or near-parturient phase, 13 subjects had their hair element levels measured again. Except for the deficiencies of calcium, iron, or/and zinc, these subjects were all healthy without symptoms of any diseases. The concentrations of hair Ca, Fe, and Zn were measured by X-ray fluorescence (XRF) spectrometry. These concentrations of the three hair elements measured at three different times were statistically analyzed. From the analyses, it was clear that hair concentrations of Ca, Fe, and Zn could reflect the effects of supplementation. Also, the mutual resistant effects among Ca-, Fe-, and Zn-nutrients were revealed. However, by appropriate combination, the mutual resistant effects could be depressed and mutual promotional effects might be enhanced. Finally, it could be concluded that mineral element deficiencies might be convalesced by adequate compensations of mineral element nutrients. PMID- 10468165 TI - Nabumetone induces less gastrointestinal mucosal changes than diclofenac retard. AB - The aim of the study was to compare the efficacy and the effects on the mucosa of the gastrointestinal tract (GIT) of nabumetone and diclofenac retard in patients with osteoarthritis (OA). An open, multicentre, randomised, comparative, endoscopy-blind parallel group study included 201 patients with nabumetone and 193 patients with diclofenac retard suffering from moderate to severe OA of the knee or hip joint. Twelve clinical efficacy variables were assessed and a portion of the population underwent gastroduodenoscopy. All patients exhibited significant improvement in pain severity and pain relief (p < 0.001 and p < 0.0001, respectively) but there were no differences between the groups for all the efficacy variables. Eleven per cent of patients on nabumetone and 19% on diclofenac experienced GIT side-effects. Sixty-nine patients with nabumetone and 61 with diclofenac underwent gastroduodenoscopy. The differences in the mucosal grade for the oesophagus, stomach and duodenum at baseline were not significant. In the oesophagus there were significantly less changes after treatment with nabumetone (p = 0.007) than with diclofenac; there were similar findings in the stomach (p < 0.001) but the difference in the duodenum was not significant. This study indicates that nabumetone and diclofenac retard have similar efficacy in the treatment of OA, but nabumetone has significantly fewer GIT side-effects. PMID- 10468166 TI - Clinical significance of antiproteinase 3 antibody positivity in cANCA-positive patients. AB - We addressed the clinical significance of antiproteinase 3 (anti-PR3) antibody (Ab) positivity by reviewing the files of 79 patients whose serum contained antineutrophil cytoplasmic antibodies with a cytoplasmic staining pattern (cANCA) and had been tested for anti-PR3 reactivity. Vasculitis was present in most (22/35) cANCA+ PR3+ patients but in only a few (5/44) cANCA+ PR3- patients, thereby suggesting that anti-PR3 Ab positivity in cANCA+ patients is more indicative of vasculitis than cANCA positivity alone. Noteworthy, one-third of cANCA+ PR3+ patients -- those with anti-PR3 Ab titres lower than 100 U/ml -- did not suffer from vasculitis. Anti-PR3 reactivity in vasculitis patients was only weakly associated with Wegener's granulomatosis (WG), as nine out of 22 cANCA+ PR3+ vasculitis patients (41%) did not fulfil the ACR classification criteria for WG. There was no correlation between anti-PR3 Ab titres and disease activity at diagnosis. However, titres measured when patients were in remission were much lower than initial values. Taken together, our results indicate that anti-PR3 Ab positivity should be interpreted in its clinical context. PMID- 10468167 TI - The program for rheumatic independent self-management: a pilot evaluation. AB - The 'Program for Rheumatic Independent Self-Management' (PRISM) is an interdisciplinary programme that integrates group education and individualised treatment using the principles of self-management, adult learning, case management and self-efficacy enhancement. This study is a before-after evaluation of 57 individuals who attended PRISM. Outcome measures were selected to measure self-efficacy, disability, pain and ability to cope. The mean self-efficacy score increased immediately following the programme and this improvement was maintained at 6-month follow-up. Disability decreased from baseline to 6-month follow-up. There was a decrease in the mean level of pain from post-class to 6-month follow up. All of these changes were statistically significant. These preliminary findings suggest that PRISM may be effective in enhancing self-efficacy, and reducing disability and pain. PMID- 10468168 TI - Autologous blood transfusion with recombinant erythropoietin treatment in anaemic patients with rheumatoid arthritis. AB - The aim of this study was to determine the conditions under which a sufficient preoperative amount of autologous blood could be obtained with administration of rHuEPO (recombinant human erythropoietin) in anaemic patients with rheumatoid arthritis (RA). Thirty-one patients (29 female, two male) with RA who were unable to donate any autologous blood owing to a haemoglobin level of less than 11 g/dl were recruited for this study. Their mean age at the time of operation was 59.3 years. The study protocol for preoperative autologous blood donations started 2.7 weeks before surgery. All patients received 6000 IU rHuEPO intravenously three times a week, supplemented with 40 mg intravenous saccharated ferric oxide at each rHuEPO administration. The protocol also included the provision that 200 g of blood at the first and third donations and 400 g of blood at the second donation were collected. The patients who were able or unable to donate 800 g of blood by this protocol were regarded as having a good or poor response, respectively, to rHuEPO. Patients with a poor response to rHuEPO showed greater clinical symptoms (morning stiffness, the number of swollen joints, Ritchie index) and higher laboratory inflammation parameters (ESR, CRP, platelets, IL-6, TNFalpha, IL-1beta) than patients with a good response to rHuEPO. The poor response group showed a significant decrease in the progression of inflammation compared with the good-response group. Before treatment with rHuEPO, anaemia in the poor-response group was the same as that in the good-response group, except for impairment of UIBC (unsaturated iron-binding capacity). The poor-response group had a higher blood loss than the good-response group. In conclusion, anaemic RA patients should be considered as candidates for aggressive blood conservation interventions that depend on erythropoietin-modulated erythropoiesis. However, it is important to determine this approach under good control of inflammation. PMID- 10468169 TI - Neurological involvement in primary Sjogren's syndrome: clinical and instrumental evaluation in a cohort of Italian patients. AB - To evaluate nervous system involvement in a cohort of Italian patients with primary Sjogren's syndrome (pSS), 87 unselected patients (83 female, and four male) observed consecutively at our institution over a period of 5 years were screened by clinical and instrumental (MRI, SPECT, electrophysiological testing, CSF analysis) investigations for peripheral and central neurological abnormalities. Seroimmunological parameters and extraglandular features other than neurological manifestations were also evaluated. Seven patients had central nervous system (CNS) disease (8%), mostly non-focal dysfunction, and 12 had peripheral nervous system (PNS) disease (13.8%), mostly mild or severe sensory or sensory-motor polyneuropathies. One patient had concomitant CNS and PNS involvement. Compared with CNS disease, PNS involvement occurred in older patients (> 50 years), independent of the disease duration. Patients with and without neurological abnormalities did not differ for seroimmunological parameters (including antiphospholipid antibodies) or extraglandular manifestations. From a statistical point of view, the only relevant finding was the detection of a slight increase in serum IgA and IgM levels (p < 0.05) in patients with an intact nervous system. Neurological involvement in pSS, be it central or peripheral, is not a rare finding. A careful clinical neurological evaluation, combined with a multiplicity of instrumental investigations, is recommended in the global assessment of pSS patients. PMID- 10468170 TI - Antineutrophil cytoplasmic antibodies in juvenile chronic arthritis. AB - We present the results of antineutrophil cytoplasmic antibody (ANCA) staining in patients with juvenile chronic arthritis (JCA). Thirty-one patients with an age range of 1-16 years were included in the study: 13, 15 and three patients, respectively, were classified having oligoarticular, polyarticular and systemic onset disease. Indirect immunofluorescence analysis revealed ANCA staining in 45% of the patients. All, except one, revealed atypical pANCA staining. ELISA studies for anti-myeloperoxidase were positive in only one patient with typical pANCA staining. PR-3 ANCA tested negative in all patients. There were no significant correlations between ANCA staining and the clinical parameters of the patients. We conclude that, although the specificity of ANCA in JCA remains to be elucidated, it may be effective in the pathogenesis of the disease. PMID- 10468171 TI - Low back pain and posture. AB - This study was designed to analyse postural aberrations of the back and lower back region in patients with acute and chronic low back pain and to investigate the accordance of clinical and radiological assessments. Fifty patients with acute and 50 with chronic low back pain and 50 controls were studied and a detailed spinal physical examination was performed. In addition, the angles of thoracic kyphosis, lumbar lordosis and sacral inclination were assessed radiologically. Differences among the three groups, correlations of radiological parameters with each other and with clinical parameters such as age, gender and body mass index, and the accordance of clinical and radiological postural assessments were investigated. It was concluded that there were no statistically significant differences among the groups for angles of thoracic kyphosis, lumbar lordosis and sacral inclination; however, we found significant correlations among all radiological parameters, especially between lumbar lordosis and sacral inclination. Both lumbar lordosis and sacral inclination were increased with body mass index, and lumbar lordosis and thoracic kyphosis were increased with age. Clinical assessments of thoracic kyphosis and lumbar lordosis were not in accordance with radiological assessments. PMID- 10468172 TI - Surgical management of calcific tendinitis of the shoulder: an analysis of 26 cases. AB - Chronic calcific tendinitis of the shoulder has good results after medical treatment in most cases. Around 10% of the patients resistant to medical treatment require surgery. We report 22 patients operated on using an open procedure. The operation included removal of the calcific deposit and an acromioplasty. The patients were rated preoperatively and postoperatively for pain, limitations in activities of daily living, range of movement and power according to a questionnaire and the Constant assessment. Globally the average score increased from 52.2 points out of 100 at the preoperative examination to 89.3 postoperatively with a mean follow up of 23 months. The most favourable results are obtained in patients with the longest interval between onset of disease and intervention (over 1 year) and with a progressive course of disease. Different techniques are discussed. Arthroscopically assisted procedures and open techniques are compared. PMID- 10468173 TI - Adhesion and lymphocyte costimulatory molecules in systemic rheumatic diseases. AB - Adhesion molecules expressed on the surface of immune cells transduce a variety of cell-activating signals and mediate important interactions by binding to multiple specific counter-receptors expressed on other cells or on extracellular matrix components. A large number of aberrations in the expression of cell-bound molecules at the mRNA and protein level in vivo have been described in patients with autoimmune connective tissue diseases. In vitro studies suggest the presence of functional abnormalities of adhesive pathways, at least at some points of the disease. Increased circulating levels of isoforms of several adhesion molecules have also been demonstrated in these patients. The possible involvement in the pathogenesis of rheumatoid arthritis, systemic lupus erythematosus, Sjogren's syndrome and systemic sclerosis of E-, P- and L-selectins, of some integrins and of several adhesion molecules of the immunoglobulin superfamily that in addition participate in lymphocyte costimulation will be discussed in this review. Further studies on migration and recruitment patterns of immune cells into inflamed tissues, as well as on possible defects of lymphocyte activation in these patients, are expected to expand our knowledge on systemic autoimmune responses and identify targets for specific immunotherapy. PMID- 10468174 TI - Panniculitis induced by collagen and plexiglas microsphere injections used for aesthetic treatment. AB - The authors report an unusual case characterised by the prompt appearance of panniculitis after injections of collagen and plexiglas microsphere for aesthetic aim and review the literature about aesthetic treatments and rheumatic disorders. PMID- 10468175 TI - Giant cell arteritis can be associated with T4-lymphocytic alveolitis. AB - We describe three patients with histologically proven giant cell arteritis who presented with respiratory complaints. In one patient, dry cough and dyspnoea dominated the clinical picture. In the other two patients, a diagnosis of giant cell arteritis was readily suspected by the presence of typical complaints, although both patients spontaneously mentioned a persistent cough and dyspnoea, respectively. Radiographs of the chest were normal. Lung function tests, including a carbon monoxide (CO)-diffusion capacity measurement, were always normal. Broncho-alveolar lavage fluid examination showed a normal cell count but an increased number of lymphocytes (16-61%) with a predominance of T4-lymphocytes (65.5-84.5%). We conclude that respiratory complaints and T4-lymphocytic alveolitis can be associated with giant cell arteritis. PMID- 10468176 TI - Idiopathic hypoparathyroidism associated with adult rheumatoid arthritis. AB - A 48-year-old female patient with adult onset idiopathic hypoparathyroidism diagnosed at the age of 28 years developed a typical seropositive rheumatoid arthritis (RA) at 46 years of age after several years of evolution of a palindromic rheumatism. Only one case of an association between idiopathic hypoparathyroidism and RA has been described in the medical literature. Autoimmunity seems to play a pivotal role in the aetiopathogenesis of both diseases, and could explain the nature of this association; nevertheless, a chance association could not be excluded. PMID- 10468177 TI - Reactive arthritis induced by Clostridium difficile enteritis as a complication of Helicobacter pylori eradication. AB - Clostridium difficile has recently been established as a cause of reactive arthritis (ReA). We present a case of Clostridium difficile-induced ReA as a complication of Helicobacter pylori eradication, which, to the best of our knowledge, is the first such case reported. PMID- 10468178 TI - Non-steroidal anti-inflammatory drugs with adverse psychiatric reactions: five case reports. AB - Adverse drug reactions of non-steroidal anti-inflammatory drugs (NSAIDs) are quite prevalent, but there are few reports about possible adverse psychiatric reactions, which may be ignored or underestimated. We describe here five psychiatric outpatients, two with major depressive disorders, one bipolar disorder, one schizophrenic disorder and one anxiety disorder, who were treated with NSAIDs for pain due to rheumatoid arthritis, osteoarthritis or other painful neuromuscular conditions. All five patients developed a moderate to severe depressive state, three patients became obviously paranoid, and four had either thoughts of suicide or an attempt while undergoing co-administration of NSAIDs. The psychiatric symptoms remitted when the NSAIDs were stopped. The depressive and paranoid symptoms returned on seven occasions of re-use or re-challenge with the same or a different type of NSAID in all five patients. When the NSAIDs were stopped again, the patients had another remission of the adverse psychiatric reactions, and eventually recovered to their baseline mental states in clear temporal relationships. The cases presented suggest that NSAIDs can induce or exacerbate idiosyncratic reproducible adverse psychiatric symptoms in certain vulnerable patients, including those with a variety of psychotic or neurotic disorders, and also in elderly persons, but these undesirable side-effects were generally transient and disappeared on withdrawal of the NSAIDs. PMID- 10468180 TI - Severe oral ulcerations induced by alendronate. AB - The authors report the case of a 54-year-old man suffering from steroid-induced osteoporosis, who presented with severe oral ulcerations due to sucking alendronate tablets instead of swallowing them. The misuse of alendronate in this case, underlies the risk of direct mucosal injury with this drug. PMID- 10468179 TI - High anti-golgi autoantibody levels: an early sign of autoimmune disease? AB - IgG anti-Golgi complex antibodies were detected by means of indirect immunofluorescence in the sera of five patients during routine investigation for suspected systemic autoimmune disease. The typical picture of paranuclear fluorescence was observed on the HEp-2 cell line and in tissue sections; anti Golgi specificity was confirmed on the HEp-2 cells using the immunoperoxidase method. The phenomenon was transient in three patients with a probable viral infection and whose sera had low titre antibodies; however, it was persistent and in high concentration in the other two who, five years later, developed an autoimmune disease. Only in the sera of these last two patients were specific bands of 123, 72, 46, 37 and 26 kilodaltons found by the immunoblotting technique on cytoplasmic extracts. Although the detection of anti-Golgi autoantibodies is rare, and may represent a transitory epiphenomenon in patients with a viral infection, their presence in high titre in the absence of a clear clinical picture may constitute an early sign of systemic autoimmune disease. PMID- 10468181 TI - Serum prolactin levels in Behcet's Syndrome. AB - Since prolactin (PRL) has been implicated as playing a role in the pathogenesis of certain autoimmune diseases and since Behcet's Syndrome (BS) is a unique systemic vasculitis, we investigated serum PRL levels in patients with BS. We found that mean PRL levels in patients with clinically active BS, were not significantly higher than patients with clinically inactive BS and healthy controls. This finding may be regarded as evidence that a contribution of hyperprolactinemia to the aetiopathogenesis of BS seems unlikely. PMID- 10468182 TI - DNA viruses and oncogenesis. AB - This review addresses the biology and role of DNA viruses in oncogenesis. PMID- 10468183 TI - Post-liver transplantation lymphoproliferative disorders with and without infusions of donor bone marrow cells. AB - Donor bone marrow cells (DBMC) infusions have been used in an attempt to decrease the untoward effects of immunosuppressive treatment and to improve immunocompetence in the post-liver transplantation (PLT) period. Between March 1987 and July 1996, 558 orthotopic liver transplantations were performed at Jackson Memorial Hospital/University of Miami. Of these, 164 patients (29%) received 10 x 10(8) DBMC/Kg using various schedules. All patients received similar immunosuppressive therapy. After a minimum follow up of 1 year, five cases of Posttransplant Lymphoproliferative Disorder (PTLD) were diagnosed in patients without DMBC (1.3%, 5/394) when compared with none (0/164) in patients who received DBMC (p = 0.15, Fisher). Four patients had malignant lymphoma and one a diffuse atypical lymphoproliferative disorder. All lymphomas were non Hodgkin's B-cell type, three diffuse large cell lymphoma, and one mixed cell lymphoma. All PTLD tested positive for EBV by in situ hybridization. Lymphomas occurred at 2, 4, 6 months and 4 years PLT. The outcome was poor with one patient diagnosed at autopsy while two patients died a few days after diagnosis. An 8 year-old girl is the only long-term survivor (> 5 years) after a partial response to combination chemotherapy and radiation therapy. The patient with diffuse atypical lymphoproliferative disorder died 3 months later. All patients with PTLD had histologic evidence of liver rejection. Although there is no statistical significant difference between the two groups, a larger cohort of patients will determine the significance of DBMC in preventing PTLD. We believe that the infusion of cytotoxic donor T cells found in the DBMC might suppress EBV-related lymphomagenesis. PMID- 10468184 TI - The immune system as anti-tumor sentinel: molecular requirements for an anti tumor immune response. AB - The concept behind immune surveillance against cancer is that tumor cells continuously develop, but that there may not be clinical evidence of their presence because the immune system recognizes the cells as foreign and destroys them. A clear role for the immune system in preventing and/or eliminating tumors is emerging as insights into the molecular requirements for the induction and effector function of cytolytic T lymphocytes (CTL) have been gained. Using murine tumor rejection models, the role of particular molecular components of the immune system in controlling tumor growth has been defined. However, tumor rejection does not always occur spontaneously in vivo, indicating that defects in the generation or execution of an anti-tumor immune response may be common. Understanding defects when they arise should allow for development of new therapeutic approaches in tumor-bearing individuals. Many clinical studies are underway to test strategies to induce or heighten an antitumor immune response in cancer patients. PMID- 10468185 TI - Interleukin-5 involvement in ovalbumin-induced eosinophil infiltration in mouse food-allergy model. AB - A number of studies demonstrating the important role of interleukin-5 (IL-5) in eosinophil infiltration were reported. Antigen-induced eosinophil infiltrations to the trachea and skin were inhibited by pretreatment with monoclonal anti-IL-5 antibody. In this study, the role of IL-5 in eosinophil infiltration to the gut by oral challenge in mice is investigated. A marked eosinophil infiltration to the lamina propria was induced by oral challenge with ovalubumin (OVA) in Balb/c mice intraperitoneally sensitized with OVA, and peaked at 6 h after the oral challenge. Intraperitoneal preadministration of monoclonal anti-IL-5 antibody significantly decreased the eosinophil infiltration to the lamina propria. Furthermore, analysis by reverse transcription polymerase chain reaction (RT-PCR) demonstrated that IL-5 mRNA expression was induced in the lamina propria in an antigen-specific manner and the expression peaked at 6 h and declined thereafter. In-situ hybridization (ISH) revealed the presence of IL-5 mRNA positive cells at lesion site. As in bronchial mucosa and skin, IL-5 may play an important role in eosinophil recruitment to the lesion site in IgE mediated gut late phase reaction. PMID- 10468187 TI - Expression of TGF-beta 1, -beta 2 and -beta 3 in localized and systemic scleroderma. AB - Scleroderma is a generalized or localized disorder which leads to fibrosis of the affected organs. TGF-beta has been implicated as a causal agent in its pathogenesis. In mammals, TGF-beta comprises a family of three members, beta 1, beta 2 and beta 3. Since cutaneous wound healing is thought to result either in formation of a scar or in scar-free tissue regeneration, depending on the relative amounts of the beta 3 isoform, the expression of all three isoforms was studied in skin biopsies of patients with either localized or systemic scleroderma. mRNA for all three isoforms was detected in inflammatory skin areas of both disease forms, but never in sclerotic or healthy skin. Immunohistochemical analysis confirmed expression of beta1 and beta 2 proteins in inflammatory skin of patients, whereas beta 3 protein appeared to be present in the subepidermal area and also found throughout the dermis of patients and healthy dermis as well. PMID- 10468186 TI - Expression of vitamin D receptor in cultured human keratinocytes and fibroblasts is not altered by corticosteroids. AB - Topical vitamin D3 therapy is one of the mainstays of psoriasis treatment. However, the effectiveness of combination therapy with topical vitamin D3 and corticosteroids is still controversial. It has been reported that topical vitamin D3 treatment following topical corticosteroids is less effective than that without preceding corticosteroid treatment. We hypothesized that vitamin D receptor (VDR) in the skin is down-regulated by topical corticosteroids. To obtain support for this hypothesis, we determined VDR protein levels in cultured keratinocytes and fibroblasts after corticosteroid treatment. VDR levels were quantified by Western blot analysis with a Fluorolmager. Keratinocytes and fibroblasts were obtained from four psoriasis patients and four normal controls. VDR levels were altered in neither normal nor psoriatic keratinocytes by 2-day incubation with dexamethasone (1x10(-9)-1x10(-6) M) or clobetasol propionate (1x10(-9)-1x10(-6) M). Similarly, VDR levels in normal and psoriatic fibroblasts were not affected by 2-day incubation with dexamethasone (1x10(-6) M). These findings suggest that down-regulation of VDR by topical corticosteroids in keratinocytes and fibroblasts of psoriasis is unlikely. PMID- 10468188 TI - High frequency of DNA aneuploidy detected by DNA flow cytometry in Bowen's disease. AB - To detect DNA aneuploidy in Bowen's disease, we investigated DNA flow cytometric analysis. Single cell suspensions were prepared from 18 fresh samples histopathologically diagnosed as solitary Bowen's disease and analyzed by DNA flow cytometry. In 16 (89%) of 18 lesions, DNA aneuploidy was demonstrated with a single aneuploid peak. DNA indices ranged from 1.29 to 1.74. The incidence of DNA aneuploidy in Bowen's disease is higher than those of cutaneous squamous cell carcinoma, which was 25-80% in the previous reports. Therefore, in Bowen's disease. DNA aneuploidy may not imply a good marker for characteristics of non melanoma skin cancer. A single aneuploid peak commonly observed in Bowen's disease suggests that this disease consists of the monoclonal proliferation of keratinocytes containing abnormal DNA content. PMID- 10468189 TI - Melanocyte melanin augments sparfloxacin-induced phototoxicity. AB - Quinolone has become one of the common causative agents of drug-induced photosensitive dermatitis, and its phototoxicity has now become a clinical issue. The mechanisms of long-lasting phototoxicity induced by sparfloxacin (SPFX) were investigated using melanotic and amelanotic melanoma cell lines. The sensitivities to SPFX and UVA irradiation up to 6 J/cm2 of both cells were not significantly different. However, the melanotic melanoma cell cultured with 1-150 microg/ml of SPFX showed significantly higher sensitivity to UVA irradiation compared with that of the amelanotic melanoma cell. By pulse culture with SPFX, the bound SPFX level of melanotic cell line was 10-20 times higher than that of the amelanotic cell line. These data strongly suggest that the melanin in melanocytes and keratinocytes accumulates the antibacterial quinolone and increases its tissue concentration in the basal epidermis and contributes to the long-lasting photo-injury and liquefaction degeneration of the basal cells in antibiotic quinolone phototoxic dermatitis. PMID- 10468190 TI - Lymphocyte stimulation test with drug-photomodified cells in patients with quinolone photosensitivity. AB - Quinolone antibacterial agents, known to elicit photosensitive dermatitis as an adverse effect, have both phototoxicity and photoallergenicity. The latter potency is mainly derived from their photohaptenic moiety; quinolones covalently bind to protein and cells upon exposure to ultraviolet A (UVA) light. Our previous study has shown the in vivo and in vitro antigenicity of quinolone photomodified cells in mice. Here, we examined the presence of sensitized lymphocytes that react with quinolone-photomodified autologous cells in patients with photosensitivity to quinolones. A flow cytometric analysis using a monoclonal antibody specific to quinolone photoadducts demonstrated that peripheral blood mononuclear cells (PBMC) were successfully photomodified with quinolones upon exposure to UVA. PBMC from quinolone-photosensitive patients were cocultured with autologous PBMC photomodified with the causative drug. Modest but significant proliferative responses of responder lymphocytes were found in patients photosensitive to lomefloxacin, fleroxacin, and enoxacin, indicating photoallergic mechanism in these patients. On the other hand, sparfloxacin photosensitive patients exhibited negative lymphocyte stimulation test, suggesting that its photosensitivity is mainly phototoxic. When UVA-preirradiated quinolones were used as stimulators, only fleroxacin exceptionally stimulated patients' PBMC, indicating its prohaptenic as well as photohaptenic properties. These findings suggest the presence of circulating sensitized T cells in patients with photosensitivity to certain quinolones. PMID- 10468191 TI - Human desmocollin 1a transiently expressed in COS-7 cells and NIH 3T3-3 cells is reacted by IgG4 antibodies in a pemphigus foliaceus serum. AB - Human desmocollin (Dsc) 1 is an autoantigen in the subcorneal pustular dermatosis type of IgA pemphigus. Moreover, Dscs, particularly bovine Dscs, are recognized by IgG antibodies in certain sera of various types of pemphigus by immunoblotting. In this study, 10 pemphigus sera were examined by immunofluorescence for IgG antibodies to human Dsc1a, Dsc2a and Dsc3a transiently expressed in COS-7 and NIH 3T3-3 cells using two different transfection methods. IgG antibodies in a number of sera showed a strong background staining with untransfected COS-7 and NIH 3T3-3 cells. Using COS-7 cells, a pemphigus foliaceus serum, which did not stain untransfected COS-7 cells, was found to contain IgG antibodies reactive exclusively with Dsc1a. This serum showed the same reactivity in studies using NIH 3T3-3 cells. Moreover, this pemphigus foliaceus serum contained IgG4, but not IgG1 antibodies, to Dscla expressed in the NIH 3T3-3 cells. These results indicate that autoantibody response in pemphigus foliaceus might be more heterogenous than hitherto supposed. PMID- 10468192 TI - Intradermal transfer of caspase-1 (CASP1) DNA into mouse dissects: role of CASP1 in interleukin-1beta associated skin inflammation and apoptotic cell death. AB - Caspase-1 (CASP1) interleukin-1beta (IL-1beta) converting enzyme (ICE) has been cloned as a specific enzyme which activates the biologically inactive pro-form of IL-1beta into biological active IL-1beta. Based on the significant homology to Ced-3, Caenorhabditis elegans apoptotic gene and, proof of apoptotic activity of ICE in rat fibroblasts, ICE was renamed as CASP1. In contrast to in vitro functions, the in vivo significance of high expression of CASP1 in skin remains to be elucidated. We transferred plasmid DNA encoding murine CASP1 with beta actin promoter into mouse skin. The CASP1 DNA-injected skin, but not skin injected with control plasmid without CASP1, developed localized erythema with subcutaneous nodules. The nodules were associated with marked inflammatory infiltrates. The apoptotic cells detected by the TUNEL method were distributed in and around the inflammatory foci. The plasma IL-1beta level of CASP1 DNA-injected mouse was elevated compared with that of the control DNA-injected mouse. These inflammatory reactions of CASP1 DNA-injected skin were suppressed by treatment with neutralizing anti-murine IL-1beta antibodies, but the TUNEL positive apoptotic cells were still detected. This study clearly demonstrate dual roles of CASP1 in causing IL-1beta associated granulomatous skin infiltration and inducing apoptotic cell death in vivo. PMID- 10468193 TI - Levels of soluble interleukin-2 receptor in the sera and skin tissue fluids of patients with vitiligo. AB - In order to study the possible role of soluble interleukin-2 receptor (sIL-2R) in the pathogenesis of vitiligo, we measured the levels of sIL-2R in the sera and the tissue fluids from skin lesions and uninvolved skin from 41 patients with vitiligo, using the method of sandwich ELISA with monoclonal and polyclonal antibodies. The results showed that the overall level of serum sIL-2R from patients with vitiligo was significantly higher than that from normal control group (P<0.01). The serum level of sIL-2R in both generalized and focal types was significantly higher than that in normal control group (P<0.01), but there was no significant difference between the segmental type and normal control group (P>0.05). The level of serum sIL-2R from patients with vitiligo in progressive stage was significantly higher than that in stable stage (P<0.01). The level of sIL-2R in the tissue fluid from the skin lesion of the patient was significantly higher than uninvolved skin of the same patient (P<0.05). The above data indicate that there is an association between the expression of sIL-2R and the pathogenesis of vitiligo. The level of serum sIL-2R in vitiligo is related to the activity of the disease, therefore the measurement of it may be significant in estimating the severity and the prognosis of the disease. PMID- 10468195 TI - Effect of the in vivo catalase inhibition on aminonucleoside nephrosis. AB - Reactive oxygen species have been involved in the pathophysiology of puromycin aminonucleoside (PAN)-nephrosis. The role of H2O2 in these rats may be studied modulating the amount or activity of catalase, which breakdowns H2O2 to water and oxygen. To explore the role of H2O2 in this experimental model, we studied the effect of the in vivo catalase inhibiton with 3-amino-1,2,4-triazole (ATZ) on the course of PAN-nephrosis. Four groups of rats were studied: control rats (CT group), PAN-injected rats (PAN group), ATZ-injected rats (ATZ group), and ATZ- and PAN-injected rats (ATZPAN group). Rats were placed in metabolic cages to collect 24 h urine along the study, ATZ (1 g/kg) was given 24 h before PAN injection (75 mg/kg), and the proteinuria was measured on days 0, 2, 4, 6, 8, and 10. Proteinuria started before (day 4) and was significantly higher on days 6, 8, and 10 in the ATZPAN group than in the PAN group. On day 10, hypercholesterolemia was significantly higher in the ATZPAN group than in the PAN group. These data indicate that the in vivo catalase inhibition magnifies PAN-nephrosis, suggesting that H2O2 is produced in vivo and involved in the renal damage in this experimental disease. PMID- 10468194 TI - Overexpression of monocyte-derived cytokines in active psoriasis: a relation to coexistent arthropathy. AB - An overexpression of inflammatory cytokines has been found in the lesional skin as well as peripheral blood in patients with psoriasis, although its etiological significance is not yet understood. In order to evaluate the cell type responsible for the elevated cytokines in the peripheral blood, we investigated cytokine profiles of the fractionated peripheral blood mononuclear cells (PBMCs) in 30 patients with psoriasis and 27 healthy controls. Without stimulation, higher levels of interleukin (IL)-1beta, IL-6, and IL-8 were produced by freshly isolated PBMCs from the patients than those from the controls. In the fractionated PBMCs, the monocyte-rich fractions were mainly responsible for the production of these cytokines and mRNA. The elevated levels of monocyte-derived cytokine mRNAs decreased following successful treatment with cyclosporin A. Although no correlation was found between the cytokine levels and the psoriasis area and severity index (PASI) scores, patients with arthropathy showed significantly high production levels of IL-1beta, IL-6, and IL-8. These findings suggest that monocytes are the major cell source producing inflammatory cytokines in the peripheral blood of psoriasis, and the increased cytokine levels are related to the coexistent arthropathy rather than the severity of cutaneous lesions. PMID- 10468196 TI - A covalent thymine-tyrosine adduct involved in DNA-protein crosslinks: synthesis, characterization, and quantification. AB - A thymine-tyrosine adduct, (3-[(1,3-dihydro-2,4-dioxopyrimidin-5-yl)methyl]-L tyrosine), was synthesized using a simple, single-step condensation between 5 (hydroxymethyl)uracil and L-tyrosine. This approach provides access to useful quantities (mg-g) of analytically pure reference material, and with minor modification, to stable isotope-labeled analogues (isotopomers). With reference material and a suitable internal standard available, isotope-dilution liquid chromatography-electrospray ionization-tandem mass spectrometry (LC/MS/MS) was used to assay the adduct in a model system purged of oxygen, i.e., a gamma irradiated N2O-saturated aqueous solution of thymine and tyrosine. The convenient synthetic route to standards and the method for quantification reported here will prove useful in assessing the significance of the adduct in biological systems. These studies also highlight the potential for artefactual adduct formation if the appropriate substrates are present under acidic conditions. PMID- 10468197 TI - Protein carbonyl formation on mucosal proteins in vitro and in dextran sulfate induced colitis. AB - Reactive oxygen and nitrogen species have been implicated as mediators of mucosal injury in inflammatory bowel disease, but few studies have investigated protein oxidation in the inflamed mucosa. In this study, protein carbonyl formation on colonic mucosal proteins from mice was investigated following in vitro exposure of homogenates to iron/ascorbate, hydrogen peroxide, hypochloric acid (HOCl), or nitric oxide (*NO). Total carbonyl content was measured spectrophotometrically by derivatization with dinitrophenylhydrazine (DNPH), and oxidation of component proteins within the tissue was examined by Western blotting for DNPH-derivatized proteins using anti-dinitrophenyl DNP antibodies. These results were compared with protein carbonyl formation found in the acutely inflamed mucosa from mice with colitis induced by dextran sulfate sodium (DSS) administered at 5% w/v in the drinking water for 7 d. In vitro, carbonyl formation was observed after exposure to iron/ascorbate, HOCl and *NO. Iron/ascorbate (20 microM/20 mM) exposure for 5 h increased carbonyl groups by 80%, particularly on proteins of 48, 75-100, 116, 131, and 142 kDa. Oxidation by 0.1 and 0.5 mM HOCl did not increase total carbonyl levels, but Western blotting revealed carbonyl formation on many proteins, particularly in the 49-95 kDa region. After exposure to 1-10 mM HOCl, total carbonyl levels were increased by 0.5 to 12 times control levels with extensive cross-linking and fragmentation of proteins rich in carbonyl groups observed by Western blotting. In mice with acute colitis induced by DSS, protein carbonyl content of the inflamed mucosa was not significantly different from control mucosa, (7.80 +/- 1.05 vs. 8.43 +/- 0.59 nmo/mg protein respectively, p = .16 n = 8, 10); however, Western blotting analysis indicated several proteins of molecular weight 48, 79, 95, and 131 kDa that exhibited increased carbonyl content in the inflamed mucosa. These proteins corresponded to those observed after in vitro oxidation of normal intestinal mucosa with iron/ ascorbate and HOCl, suggesting that both HOCl and metal ions may be involved in protein oxidation in DSS-induced colitis. Identification and further analysis of the mucosal proteins susceptible to carbonyl modification may lead to a better understanding of the contribution of oxidants to the colonic mucosa tissue injury in inflammatory bowel disease. PMID- 10468198 TI - Endogenous catechol thioethers may be pro-oxidant or antioxidant. AB - Increased catechol thioether formation is associated with Parkinson's disease. In this study, we examined whether catechol thioethers, having a lower oxidation potential than their parent catechols, would cause greater oxidative damage than their parent catechols. We synthesized 5'-S-glutathionyl, cysteinyl, and N acetylcysteinyl derivatives of dopamine and dopac, encompassing the known catechol thioethers of the mercapturate pathway. Cyclic voltametry studies showed that catechol thioethers had higher reduction potentials than their parent catechols. A higher reduction potential did not correlate with an increase in oxidative damage, measured by metal-catalyzed DNA strand breakage. 5'-S Glutathionyldopamine and the cysteinyl adducts of dopamine and dopac mediated less oxidative damage than their parent catechols. In contrast, both N acetylcysteinyl analogs were equipotent to dopamine. Oxygen consumption corresponded to DNA damage except for 5'-S-glutathionyldopamine. The glutathionyl and cysteinyl adducts of dopamine inhibited dopamine-mediated DNA damage indicating that these adducts may have antioxidant properties. 5'-S Glutathionyldopamine potentiated H2O2-mediated damage whereas 5-S cysteinyldopamine was inhibitory. Our results show that the ability of catechol thioethers to cause oxidative damage in vitro is not based simply upon the reduction potential but rather, reflects a complex relationship among structures of the parent catechol and thiol adduct, metal catalyst, and oxidant. PMID- 10468199 TI - Urinary excretion of hydroxycinnamates and flavonoids after oral and intravenous administration. AB - The urinary recoveries of the hydroxycinnamates, ferulic acid (3-methoxy, 4 hydroxy cinnamic acid), and chlorogenic acid (the quinic acid ester of 3,4 dihydroxycinnamic acid), and three structurally related flavonoids were studied in the rat. For the latter, the aglycone quercetin was compared with its 3 glucoside (isoquercitrin) and 3-rhamnoglucoside (rutin). Doses of 50 mg/kg were administered via the oral and intravenous routes and urine collected over the subsequent 24-h period. Reverse phase HPLC with photo-diode array detection was used to analyze the unchanged compound and their metabolites excreted in the urine. Ferulic acid and isoquercitrin were orally absorbed (5.4 and 0.48% of administered dose, respectively) and are therefore bioavailable. In contrast, neither unchanged chlorogenic acid, rutin, quercetin, nor the conjugated metabolites in the form of glucuronide or sulphate were detected in the urine after oral dosing. All the flavonoids studied produced low total urinary recoveries after intravenous administration, 9.2% for quercetin-3 rhamnoglucoside, 6.7% for the 3-glucoside, and 2.4% for the aglycone, indicating that extensive metabolism to low molecular weight compounds or excretion via other routes may be occurring. Overall it can be stated that renal excretion is not a major pathway of elimination for intact flavonoids and hydroxycinnamates in the rat. PMID- 10468200 TI - Effect of age and caloric restriction on bleomycin-chelatable and nonheme iron in different tissues of C57BL/6 mice. AB - The objective of this study was to test the hypothesis that the widely observed age-associated increase in the amounts of macromolecular oxidative damage is due to an elevation in the availability of redox-active iron, that is believed to catalyze the scission of H2O2 to generate the highly reactive hydroxyl radical. Concentrations of bleomycin-chelatable iron and nonheme iron were measured in various tissues and different regions of the brain of mice fed on ad libitum (AL) or a calorically restricted (to 60% of AL) diet at different ages. The concentrations of these two pools of iron varied markedly as a function of tissue, age, and caloric intake. There was no consistent ratio between the amounts of nonheme and the bleomycin-chelatable iron pools across these conditions. Nonheme iron concentration increased with age in the liver, kidney, heart, striatum, hippocampus, midbrain and cerebellum of AL animals, whereas bleomycin-chelatable iron increased significantly with age only in the liver. Amounts of both nonheme and bleomycin-chelatable iron remained unaltered during aging in the cerebral cortex and hindbrain of AL mice. Caloric restriction had no effect on iron concentration in the brain or heart, but caused a marked increase in the concentration of both bleomycin-chelatable and nonheme iron in the liver and the kidney. The results do not support the hypothesis that accumulation of oxidative damage with age, or its attenuation by CR, are associated with corresponding variations in redox-active iron. PMID- 10468201 TI - Irradiation of titanium dioxide generates both singlet oxygen and superoxide anion. AB - Although photoexcited TiO2 has been known to initiate various chemical reactions, such as the generation of reactive oxygen species, precise mechanism and chemical nature of the generated species remain to be elucidated. The present work demonstrates the generation of singlet oxygen by irradiated TiO2 in ethanol as measured by ESR spectroscopy using 2,2,6,6-tetramethyl-4-piperidone (4-oxo-TMP) as a 1O2-sensitive trapping agent. Under identical conditions, the superoxide ion was also detected by spin trapping agent 5,5-dimethyl-pyrroline-N-oxide (DMPO). Kinetic analysis in the presence of both 4-oxo-TMP and DMPO revealed that singlet oxygen is produced directly at the irradiated TiO2 surface but not by a successive reaction involving superoxide anion. The basis for this view is the fact that DMPO added in the mixture increased the signals responsible for 4-oxo 2,2,6,6-tetramethyl-1-piperidinyloxy (4-oxo-TEMPO), a reaction product of 4-oxo TMP and 1O2. The detailed mechanism for the generation of 1O2 and superoxide ion by irradiated TiO2 and reactions between these species and DMPO are discussed. PMID- 10468202 TI - Photogeneration of 3beta-hydroxy-5alpha-cholest-6-ene-5-hydroperoxide in rat skin: evidence for occurrence of singlet oxygen in vivo. AB - We identified singlet oxygen adduct of cholesterol, 3beta-hydroxy-5alpha-cholest 6-ene-5-hydroperoxide (5alpha-OOH), in skin of rats pretreated with oral doses of pheophorbide a and subsequent visible irradiation, that have been known to induce photosensitive diseases in animals and humans. In a living animal body, this is the first demonstration of presence of 5alpha-OOH, that is known to be formed exclusively by reaction in vitro between singlet oxygen and cholesterol. By the quantitative determination with high performance liquid chromatography equipped with a chemiluminescence detector, we observed time-dependent increase in concentrations of 5alpha-OOH in skin of rats pretreated with oral doses of pheophorbide a and subsequent visible irradiation, suggesting the occurrence of a labile activated oxygen species, singlet oxygen, in this system. PMID- 10468203 TI - Oral administration of RAC-alpha-lipoic acid modulates insulin sensitivity in patients with type-2 diabetes mellitus: a placebo-controlled pilot trial. AB - Alpha-lipoic acid (ALA), a naturally occuring compound and a radical scavenger was shown to enhance glucose transport and utilization in different experimental and animal models. Clinical studies described an increase of insulin sensitivity after acute and short-term (10 d) parenteral administration of ALA. The effects of a 4-week oral treatment with alpha-lipoic acid were evaluated in a placebo controlled, multicenter pilot study to determine see whether oral treatment also improves insulin sensitivity. Seventy-four patients with type-2 diabetes were randomized to either placebo (n = 19); or active treatment in various doses of 600 mg once daily (n = 19), twice daily (1200 mg; n = 18), or thrice daily (1800 mg; n = 18) alpha-lipoic acid. An isoglycemic glucose-clamp was done on days 0 (pre) and 29 (post). In this explorative study, analysis was done according to the number of subjects showing an improvement of insulin sensitivity after treatment. Furthermore, the effects of active vs. placebo treatment on insulin sensitivity was compared. All four groups were comparable and had a similar degree of hyperglycemia and insulin sensitivity at baseline. When compared to placebo, significantly more subjects had an increase in insulin-stimulated glucose disposal (MCR) after ALA treatment in each group. As there was no dose effect seen in the three different alpha-lipoic acid groups, all subjects receiving ALA were combined in the "active" group and then compared to placebo. This revealed significantly different changes in MCR after treatment (+27% vs. placebo; p < .01). This placebo-controlled explorative study confirms previous observations of an increase of insulin sensitivity in type-2 diabetes after acute and chronic intravenous administration of ALA. The results suggest that oral administration of alpha-lipoic acid can improve insulin sensitivity in patients with type-2 diabetes. The encouraging findings of this pilot trial need to be substantiated by further investigations. PMID- 10468204 TI - Oxidation of extracellular matrix modulates susceptibility to degradation by the mesangial matrix metalloproteinase-2. AB - Protein oxidation occurs in aging and in various inflammatory conditions. Glomerulosclerosis is characterized by an accumulation of extracellular matrix (ECM) proteins and a paucity of glomerular mesangial cells and can be seen as an end-result of glomerular injury and in aging. ECM accumulation is the net result of the balance between synthesis and degradation. ECM may become oxidized as a part of inflammatory renal injury and with aging. We evaluated the hypothesis that oxidation of mesangial ECM could alter its susceptibility to the action of ECM degrading enzymes. Radiolabeled mesangial ECM was generated by growing cells on tissue culture plastic and incubating with [3H]proline. After removal of cells, leaving behind ECM, selected wells were oxidized using a FeCl3/EDTA/ascorbate system or treated under control conditions. The control and oxidized matrices were then incubated with concentrated supernatants from mesangial cells containing the major mesangial ECM degrading enzyme, the matrix metalloproteinase-2, whose activity was confirmed by gelatin substrate zymography. Counts released corresponding with ECM degraded were measured. ECM oxidized with this system was significantly less susceptible to degradation compared to control ECM. To confirm that this effect was specifically due to oxidative modification of the ECM rather than changes unrelated to oxidation we coincubated ECM with the oxidizing system plus the radical spin trap N-tert-butyl alpha-phenylnitrone (PBN). PBN treatment was able to prevent the impaired susceptibility to degradation induced by exposure to the oxidizing system. Exposure of ECM to milder oxidative stress, however, modestly enhanced susceptibility to degradation. These data suggest that oxidation of mesangial ECM can modulate its susceptibility to degradation. This may account for the development of ECM accumulation and glomerulosclerosis in inflammatory renal injury and in aging. PMID- 10468205 TI - Reactivity of biologically important thiol compounds with superoxide and hydrogen peroxide. AB - The reactivities of glutathione, cysteine, cysteamine, penicillamine, N acetylcysteine, dithiothreitol and captopril with superoxide generated from xanthine oxidase and hypoxanthine, and with reagent hydrogen peroxide, have been investigated. Rates of thiol loss on adding hydrogen peroxide, and superoxide dependent thiol loss and oxygen uptake were measured. The relative reactivities of the different thiols with both oxidants were inversely related to the pK of the thiol group, such that at pH 7.4, penicillamine was the most reactive. N acetylcysteine weakly reactive and no reaction was seen with captopril. For hydrogen peroxide, the calculated rate constants for the reaction with the thiolate anion all fell within the range 18-26 M(-1) s(-1). With superoxide, our results are consistent with each thiol reacting via a short chain that consumes oxygen and regenerates superoxide. Only with some of the thiols, was the consumed oxygen recovered as hydrogen peroxide. Reported values for the rate constant for the reaction of thiols with superoxide vary over four orders of magnitude, with the highest being > 10(5) M(-1) s(-1). Due to the complexity of the chain reaction, no study so far has been able to obtain accurate values and we consider the best estimates to be in the 30 to 1000 M(-1) s(-1) range. PMID- 10468206 TI - Trapping of free radicals with direct in vivo EPR detection: a comparison of 5,5 dimethyl-1-pyrroline-N-oxide and 5-diethoxyphosphoryl-5-methyl-1-pyrroline-N oxide as spin traps for HO* and SO4*-. AB - To spin trap hydroxyl radical (HO*) with in vivo detection of the resultant radical adducts, the use of two spin traps, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and 5-diethoxyphosphoryl-5-methyl-1-pyrroline-N-oxide (DEPMPO) (10 mmol/kg) has been compared. In mice treatment with 5-aminolevulinic acid and Fe3+ resulted in detection of adducts of hydroxyl radicals (HO*), but only with use of DEPMPO. Similarly, 'HO* adducts' generated via nucleophilic substitution of SO4*- adducts formed in vivo could be observed only when using DEPMPO as the spin trap. The reasons for the differences observed between DEPMPO and DMPO are likely due to different in vivo lifetimes of their hydroxyl radical adducts. These results seem to be the first direct in vivo EPR detection of hydroxyl radical adducts. PMID- 10468207 TI - Comparative study on dynamics of antioxidative action of alpha-tocopheryl hydroquinone, ubiquinol, and alpha-tocopherol against lipid peroxidation. AB - Alpha-tocopheryl quinone is a metabolite of alpha-tocopherol (TOH) in vivo. The antioxidant action of its reduced form, alpha-tocopheryl hydroquinone (TQH2), has received much attention recently. In the present study, the antioxidative activity of TQH2 was studied in various systems in vitro and compared with that of ubiquinol-10 (UQH2) or TOH to obtain the basic information on the dynamics of the antioxidant action of TQH2. First, their hydrogen-donating abilities were investigated in the reaction with galvinoxyl, a stable phenoxyl radical, and TQH2 was found to possess greater second-order rate constant (1.0 x 10(4) M(-1) s(-1)) than UQH2 (6.0 x 10(3) M(-1) s(-1)) and TOH (2.4 x 10(3) M(-1) s(-1)) at 25 degrees C in ethanol. The stoichiometric numbers were obtained as 1.9, 2.0, and 1.0 for TQH2, UQH2, and TOH, respectively, in reducing galvinoxyl. Second, their relative reactivities toward peroxyl radicals were assessed in competition with N,N'-diphenyl-p-phenylenediamine (DPPD) and found to be 6.0 (TQH2), 1.9 (UQH2), and 1.0 (TOH). Third, their antioxidant efficacies were evaluated in the oxidation of methyl linoleate in organic solvents and in aqueous dispersions. The antioxidant potency decreased in the order TOH > UQH2 > TQH2, as assessed by either the extent of the reduction in the rate of oxidation or the duration of inhibition period. The reverse order of their reactivities toward radicals and their antioxidant efficacies was interpreted by the rapid autoxidation of TQH2 and UQH2, carried out by hydroperoxyl radicals. Although neither TQH2 nor UQH2 acted as a potent antioxidant by itself, they acted as potent antioxidants in combination with TOH. TQH2 and UQH2 reduced alpha-tocopheroxyl radical to spare TOH, whereas TOH suppressed the autoxidation of TQH2 and UQH2. In the micelle oxidation, the antioxidant activities of TQH2, UQH2, and TOH were similar, whereas 2,2,5,7,8-pentamethyl-6-chromanol exerted much more potent efficacy than TQH2, UQH2, or TOH. These results clearly show that the antioxidant potencies against lipid peroxidation are determined not only by their chemical reactivities toward radicals, but also by the fate of an antioxidant-derived radical and the mobility of the antioxidant at the microenvironment. PMID- 10468208 TI - Nitric oxide-forming reactions of the water-soluble nitric oxide spin-trapping agent, MGD. AB - The objective of this study was to elucidate the nitric oxide-forming reactions of the iron-N-methyl-D-glucamine dithiocarbamate (Fe-MGD) complex from the nitrogen-containing compound hydroxyurea. The Fe2+(MGD)2 complex is commonly used in electron paramagnetic resonance (EPR) spectroscopic detection of NO both in vivo and in vitro. The reaction of Fe2+(MGD)2 with NO yields the resultant NO Fe2+(DETC)2 complex, which has a characteristic triplet EPR signal. It is widely believed that only NO reacts with Fe2+(MGD)2 to form the NO-Fe2+(MGD)2 complex. In this report, the mechanism leading to the formation of NO-Fe2+(MGD)2 was investigated using oxygen-uptake studies in conjunction with the EPR spin trapping technique. We found that the air oxidation of Fe2+(MGD)2 complex results in the formation of the Fe3+(MGD)3 complex, presumably concomitantly with superoxide (O3*-). Dismutation of superoxide forms hydrogen peroxide, which can subsequently reduce Fe3+(MGD)3 back to Fe2+(MGD)2. The addition of NO to the Fe3+(MGD)3 complex resulted in the formation of the NO-Fe2+(MGD)2 complex. Hydroxyurea is not considered to be a spontaneous NO donor, but has to be oxidized in order to form NO. We present data showing that in the presence of oxygen, Fe2+(MGD)2 can oxidize hydroxyurea to yield the stable NO-Fe2+(MGD)2 complex. These results imply that hydroxyurea can be oxidized by reactive oxygen species that are formed from the air oxidation of the Fe2+(MGD)2 complex. Formation of the NO-Fe2+(MGD)2 complex in this case could erroneously be interpreted as spontaneous formation of NO from hydroxyurea. The chemistry of the Fe2+(MGD)2 complexes in aerobic conditions must be taken into account in order to avoid erroneous conclusions. In addition, the use of these complexes may contribute to the overall oxidative stress of the system under investigation. PMID- 10468209 TI - A novel 19F-NMR method for the investigation of the antioxidant capacity of biomolecules and biofluids. AB - A new assay for the measurement of the antioxidant capacity of biomolecules by high resolution 19F-NMR spectroscopy is presented here. This method is based on the use of trifluoroacetanilidic detectors, namely trifluoroacetanilide, N-(4 hydroxyphenyl)-trifluoroacetamide and 2-hydroxy-4-trifluoroacetamidobenzoic acid. Upon hydroxyl radical attack, such fluorinated detectors yield trifluoroacetamide and trifluoroacetic acid that can be quantitatively determined by 19F-NMR spectroscopy. Trifluoroacetamide was found to be a reliable reporter of hydroxyl radical attack on the fluorinated detectors, whereas N-(4-hydroxyphenyl) trifluoroacetamide was found to be the most sensitive detector amongst the ones considered. Therefore, N-(4-hydroxyphenyl)-trifluoroacetamide has been used in competition experiments to assess the antioxidant capacity of a number of low and high molecular weight antioxidants. The antioxidant capacity of a given compound has been scaled in terms of an adimensional parameter, kF, that represents the ratio between the scavenger abilities of the fluorinated detector and the competitor. kF values obtained for low-molecular-mass compounds fall in the range 0.17 < kF < 1.5 and are in good agreement with second order rate constants (k2OH) for the reaction of the antioxidant with hydroxyl radicals. The kF value for serum albumin is much larger (46.9) than that predicted from the reported k2OH value. This finding supports the view that the protein can very effectively scavenge hydroxyl radicals as well as secondary radicals. Human blood serum showed that its antioxidant capacity is even higher than that shown by aqueous solutions of albumin at physiologic concentration suggesting a further contribution from other macromolecular serum components. PMID- 10468210 TI - Population doubling time, phosphatase activity, and hydrogen peroxide generation in Jurkat cells. AB - Differences in growth characteristics, phosphatase activity, and hydrogen peroxide generation in two clones of a T-cell leukemic line are described in this communication. Wurzburg cells had significantly shorter population doubling times compared with the parental Jurkat cells (16.6 +/- 2.0 h and 20.7 +/- 2.2 h, respectively; mean +/- SD, p < .0001, n = 20). In addition, total phosphatase activity was significantly decreased (p < .006) and hydrogen peroxide production was significantly increased (p < .002) in Wurzburg cells compared to Jurkat cells. That the cell line with the faster growth rate should have these latter two properties is entirely consistent with the positive effects of increased kinase activity and hydrogen peroxide on proliferative cellular responses in T cells. As originally described, Wurzburg cells were distinguished from Jurkat cells by their lack of CD45, a membrane protein tyrosine phosphatase, and their positive response to hydrogen peroxide-stimulation of NF-kappaB activation. We propose that these two clones, with their distinguishing characteristics, can be used to advantage in experiments designed to study the effects of antioxidants on signaling pathways that control cell life and death. PMID- 10468211 TI - The effect of experimental conditions on the levels of oxidatively modified bases in DNA as measured by gas chromatography-mass spectrometry: how many modified bases are involved? Prepurification or not? AB - Recently, an artifactual formation of a number of modified DNA bases has been alleged during derivatization of DNA hydrolysates to be analyzed by gas chromatography-mass spectrometry (GC-MS). These modified bases were 8 hydroxyguanine (8-OH-Gua), 5-hydroxycytosine (5-OH-Cyt), 8-hydroxyadenine (8-OH Ade), 5-hydroxymethyluracil (5-OHMeUra), and 5-formyluracil, which represent only a small percentage of more than 20 modified DNA bases that can be analyzed by GC MS. However, relevant papers reporting the levels of these modified bases in DNA of various sources have not been cited, and differences in experimental procedures have not been discussed. We investigated the levels of modified bases in calf thymus DNA by GC-MS using derivatization at three different temperatures. The results obtained with GC/isotope-dilution MS showed that the levels of 5-OH Cyt, 8-OH-Ade, 5-OH-Ura, and 5-OHMeUra were not affected by increasing the derivatization temperature from 23 degrees C to 120 degrees C. The level of 8-OH Gua was found to be higher at 120 degrees C. However, this level was much lower than those reported previously. Formamidopyrimidines were readily analyzed in contrast to some recent claims. The addition of trifluoroacetic acid (TFA) adversely affected the levels of pyrimidine-derived lesions, suggesting that TFA is not suitable for simultaneous measurement of both pyrimidine- and purine derived lesions. The data obtained were also compared with those previously published. Our data and this comparison indicate that no artifactual formation of 5-OH-Cyt, 8-OH-Ade, and 5-OHMeUra occurred under our experimental conditions in contrast to recent claims, and no prepurification of DNA hydrolysates by a tedious procedure is necessary for accurate quantification of these compounds. The artifactual formation of 8-OH-Gua can be eliminated by derivatization at room temperature for at least 2 h, without the use of TFA. The results in this article and their comparison with published data indicate that different results may be obtained in different laboratories using different experimental conditions. The data obtained in various laboratories should be compared by discussing all relevant published data and scientific facts, including differences between experimental conditions used in different laboratories. PMID- 10468212 TI - Black tea increases the resistance of human plasma to lipid peroxidation in vitro, but not ex vivo. AB - A number of in vitro studies have shown that polyphenols and flavonoids in tea exert significant antioxidant activity. However, epidemiologic and experimental studies have produced conflicting results. The purpose of the present study was to compare the antioxidant activity of black tea in vitro with that ex vivo. Black tea polyphenols (BTP), black tea extract (BTE), or their major polyphenolic antioxidant constituent, epigallocatechin gallate (EGCG), were added to human plasma and lipid peroxidation was induced by the water-soluble radical generator 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH). Following a lag phase, lipid peroxidation was initiated and occurred at a rate that was lowered in a dose-dependent manner by BTP. Similarly, EGCG and BTE added to plasma in vitro strongly inhibited AAPH-induced lipid peroxidation. The lag phase preceding detectable lipid peroxidation was due to the antioxidant activity of endogenous ascorbate, which was more effective at inhibiting lipid peroxidation than the tea polyphenols and was not spared by these compounds. In contrast, when eight healthy volunteers consumed the equivalent of six cups of black tea, the resistance of their plasma to lipid peroxidation ex vivo did not increase over the next 3 h. These data suggest that, despite antioxidant efficacy in vitro, black tea does not protect plasma from lipid peroxidation in vivo. The striking discrepancy between the in vitro and ex vivo data is most likely explained by the insufficient bioavailability of tea polyphenols in humans. PMID- 10468213 TI - Efficacy of the antioxidant ebselen in experimental uveitis. AB - Inflammation results in the production of free radicals. In a model of experimental uveitis upon subcutaneous injection of endotoxin to Lewis rats, i.e., endotoxin-induced experimental uveitis (EIU), we have evaluated the status of the antioxidant capacity of ocular tissues. EIU results in a decrease of glutathione (GSH) content and glutathione peroxidase (GPx) activity in whole eye homogenates 24-h after endotoxin administration. Furthermore, an increase in malondialdehyde (MDA) content was observed in these same samples, thus confirming the involvement of oxidative stress in the pathophysiology of the process. In view of the ability of the antioxidant ebselen as GPx enzyme mimic, we tested the effect of the oral treatment with two doses of 100 mg/kg body weight of ebselen (first dose administered at the same time of endotoxin, and the second after 12 h). Ebselen administration normalized the GSH and MDA contents and protected the GPx activity of the EIU rat eyes. The GPx activity in the eye homogenate of the treated rats could be completely acounted for by the ebselen-dependent GPx-like activity, i.e., GPx activity measured in the acidic supernatant of the homogenate after neutralization. Unmodified ebselen was detected in whole eye homogenates, thus it shows for the first time the penetration of ebselen through the blood aqueous and blood-retina barrier. The results herein may allow the proposal of ebselen as a suitable antiinflammatory agent in ocular tissues. PMID- 10468214 TI - Contraction of human airways by oxidative stress protection by N-acetylcysteine. AB - We examined the in vitro effects of tert-butylhydroperoxide (tBu-OOH) in human bronchial muscle. tert-Butylhydroperoxide produced concentration-dependent contractions of bronchial rings (maximum effect was 56.5 +/- 9.6% of contraction by 1 mM acetylcholine; effective concentration 50% was approximately 100 microM). tert-Butylhydroperoxide (0.5 mM)-induced contraction was enhanced by epithelial removal but abolished by indomethacin (cyclooxygenase inhibitor) and zileuton (lipoxygenase inhibitor). tert-Butylhydroperoxide produced a transient rise in intracellular calcium in human cultured airway smooth muscle cells (HCASMC). The bronchial reactivity to acetylcholine and histamine was not altered by tBu-OOH. In HCASMC, tBu-OOH (0.5 mM, 30 min) increased malondialdehyde levels (MDA; from 7.80 +/- 0.83 to 26.82 +/- 1.49 nmol mg(-1) protein), accompanied by a decrease of reduced glutathione (GSH; from 16.7 +/- 2.6 to 6.9 +/- 1.9 nmol mg(-1) protein) and an increase of oxidized glutathione (from 0.09 +/- 0.03 to 0.18 +/- 0.03 nmol mg(-1) protein). N-acetylcysteine (0.3 mM) inhibited by approximately 60% the bronchial contraction resulting from tBu-OOH (0.5 mM) and protected cultured cells exposed to tBu-OOH (MDA was lowered to 19.51 +/- 1.19 nmol mg(-1) protein, and GSH content was replenished). In summary, tBu-OOH caused contraction of human bronchial muscle mediated by release of cyclo-oxygenase and lipoxygenase products without producing airways hyperreactivity. N-acetylcysteine decreases tBu-OOH-induced contraction and protects human cultured airway smooth muscle cells exposed to tBu-OOH. PMID- 10468215 TI - Persistent oxidative stress in human colorectal carcinoma, but not in adenoma. AB - Few studies have been conducted focusing on a potential role of reactive oxygen species in tumor cell metabolism. Here we studied human colorectal adenocarcinomas and adenomas to determine whether oxidative stress is imposed on cancer cells in vivo and used specific antibodies against 8-hydroxy-2' deoxyguanosine (8-OHdG), 4-hydroxy-2-nonenal (HNE)-modified proteins, and 3-nitro L-tyrosine (3-NT) to determine whether there is an association between oxidative stress and cellular proliferation. Higher levels of oxidative modifications in DNA and proteins were observed in carcinoma cells, but not in adenoma cells, than in the corresponding nontumorous epithelial cells by immunohistochemistry as well as high-performance liquid chromatography (HPLC)-based 8-OHdG determination. The fraction of proliferating cell nuclear antigen-positive cells was proportionally associated in adenocarcinomas with the staining intensities of 8-OHdG and 3-NT. Furthermore, Western blot analysis of the proteins extracted from carcinoma cells revealed several specific proteins modified by HNE or peroxynitrite. Thus we concluded that colorectal carcinoma, but not adenoma cells, are exposed to more oxidative stress than their corresponding nontumorous epithelial cells, regardless of clinical stage and histology, and further that the oxidative stress in carcinoma cells might stimulate cellular proliferation. PMID- 10468216 TI - Activation of the superoxide-generating NADPH oxidase of intestinal lymphocytes produces highly reactive free radicals from sulfite. AB - The objective of this study was to investigate the ability of immune cells of the small intestine to produce highly reactive free radicals from the food additive sulfites. These free radicals were characterized with a spin-trapping technique using the spin traps 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide (DEPMPO) and 5,5-dimethyl-1-pyrroline N-oxide (DMPO). In the presence of glucose, purified lymphocytes from intestinal Peyer's patches (PP) and mesenteric lymph nodes (MLN) were stimulated with phorbol 12-myristate 13-acetate (PMA) to produce superoxide and hydroxyl DEPMPO radical adducts. The formation of these adducts was inhibited by superoxide dismutase or diphenyleneiodonium chloride, indicating that these cells produced superoxide radical during reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation. With the treatment of sodium sulfite, PMA-stimulated PP lymphocytes produced a DEPMPO-sulfite radical adduct and an unknown radical adduct. When DEPMPO was replaced with DMPO, DMPO-sulfite and hydroxyl radical adducts were detected. The latter adduct resulted from DMPO oxidation by sulfate radical, which was capable of oxidizing formate or ethanol. Oxygen consumption rates were further increased after the addition of sulfite to PMA-stimulated lymphocytes, suggesting the presence of sulfiteperoxyl radical. Taken together, oxidants generated by stimulated lymphocytes oxidized sulfite to sulfite radical, which subsequently formed sulfiteperoxyl and sulfate radicals. The latter two radicals are highly reactive, contributing to increased oxidative stress, which may lead to sulfite toxicity, altered functions in intestinal lymphocytes, or both. PMID- 10468217 TI - Antioxidant protection from solar-simulated radiation-induced suppression of contact hypersensitivity to the recall antigen nickel sulfate in human skin. AB - Solar radiation induces suppression of the local effector mechanisms involved in immune responses to recall antigens. By using a low-dose solar-simulated radiation protocol, we investigated whether oral supplementation of the antioxidants RRR-alpha-tocopherol combined with L-ascorbic acid prevented radiation-induced suppression of the contact hypersensitivity response to nickel sulfate. In a prospective, randomized study, nickel-sensitive individuals were given RRR-alpha-tocopherol 2 g/d oral supplements combined with L-ascorbic acid 3 g/d for 50 d (group 1). Individuals in the control group were given a placebo (group 2). The reaction to a standardized patch test with serial dilutions of nickel sulfate and the irritant skin reaction to sodium lauryl sulfate were assessed by visual grading and by reflectance spectrophotometry in radiation exposed and nonexposed skin 50 days after supplementation. Results showed that the contact hypersensitivity response to the recall antigen nickel sulfate was significantly suppressed in the radiation-exposed skin of those who took the placebo. Supplementation with RRR-alpha-tocopherol combined with L-ascorbic acid significantly protected against the radiation-induced suppression of the contact hypersensitivity response to nickel sulfate. The irritant reaction to sodium lauryl sulfate was not suppressed by radiation, and antioxidant supplementation did not modulate this response. In conclusion, a combination therapy of systemic high-dose RRR-alpha-tocopherol combined with L-ascorbic acid prevented solar simulated radiation-induced suppression of the local immune response to the recall antigen nickel sulfate in human skin. This immunoprotective effect of combined RRR-alpha-tocopherol and L-ascorbic acid could be exploited for the prevention of solar radiation-induced skin cancer in an antioxidant intervention study. PMID- 10468218 TI - Peripheral markers of oxidative stress in Parkinson's disease. The role of L DOPA. AB - Oxidative stress plays a central role in the pathogenesis of Parkinson's disease (PD). L-DOPA, the gold standard in PD therapy, may paradoxically contribute to the progression of the disease because of its pro-oxidant properties. The issue, however, is controversial. In this study, we evaluated peripheral markers of oxidative stress in normal subjects, untreated PD patients and PD patients treated only with L-DOPA. We also measured platelet and plasma levels of L-DOPA, 3-O-methyldopa (the long-lasting metabolite of the drug), and dopamine. We found that isolated platelets of treated PD patients form higher amounts of 2,3 dihydroxybenzoate, an index of hydroxyl radical generation, than platelets of controls or untreated patients. In treated patients, platelet levels of 2,3 dihydroxybenzoate were positively correlated with platelet levels of L-DOPA, 3-O methyldopa, and with the score of disease severity. Disease severity was correlated with platelet and plasma levels of L-DOPA, as well as with the daily intake of the drug. No significant differences in platelet levels of cytosolic and mitochondrial isoforms of the antioxidant enzyme superoxide dismutase were found between PD patients, either treated or untreated, and controls. Our findings lend further support to the hypothesis that L-DOPA might promote free radical formation in PD patients. PMID- 10468219 TI - Low concentrations of 17beta-estradiol reduce oxidative modification of low density lipoproteins in the presence of vitamin C and vitamin E. AB - Micromolar concentrations of estradiol are required to inhibit the oxidation of low-density lipoproteins (LDL) in vitro. Recent evidence suggests that estradiol must be modified before it can become an effective antioxidant at physiological levels. Our aim was to determine other possible conditions under which low concentrations of 17beta-estradiol can reduce LDL oxidation. LDL susceptibility to oxidation was monitored by measurements of conjugated diene formation. High levels of 17beta-estradiol reduced oxidative modification of LDL. Vitamin C and vitamin E also increased LDL resistance to Cu2+-mediated oxidation. More importantly, 10 nM 17beta-estradiol, which on its own had no effect, exhibited significant antioxidant actions in the presence of either vitamins C or E. In conclusion, supraphysiological concentrations of 17beta-estradiol are required to exert antioxidant effects directly in vitro. However, in the presence of vitamins C and E, concentrations of 17beta-estradiol close to physiological levels can also protect LDL from oxidation. PMID- 10468220 TI - EPR imaging for in vivo analysis of the half-life of a nitroxide radical in the hippocampus and cerebral cortex of rats after epileptic seizures. AB - Recently, we developed an in vivo temporal electron paramagnetic resonance (EPR) imaging technique to be applied to the brain of a rat, into which a blood-brain barrier (BBB)-permeable nitroxide radical, 3-methoxycarbonyl-2,2,5,5 tetramethylpyrrolidine-1-oxyl (PCAM) was injected intraperitoneally. This imaging technique made it possible to measure decay rates of a nitroxide radical in multiple regions of the brain simultaneously. Using this technique, the half-life of PCAM was estimated from the exponential decay of the signal intensity derived from the temporal EPR images in the hippocampus and cerebral cortex of rats after a kainic acid (KA)-induced seizure. The hippocampal half-life of PCAM after KA induced seizures was significantly prolonged (p < .01), whereas the prolongation of the cortical half-life was not significant. These findings suggest that following a KA-seizure, the intrahippocampal ability to reduce the nitroxide radical is impaired, but the ability is intact in the cerebral cortex. This is the first in vivo quantitative EPR imaging study that has a bearing on the pathogenesis of KA-induced seizures in the brain. PMID- 10468221 TI - Constitutive differences in antioxidant defense status distinguish alloxan resistant and alloxan-susceptible mice. AB - Alloxan (AL), a potent generator of superoxide and hydroxyl radicals, selectively destroys rodent pancreatic beta-cells. Alloxan-susceptible (ALS/Lt) and AL resistant (ALR/Lt) are inbred mouse strains derived in Japan by inbreeding CD-1 (ICR) mice with concomitant selection for high or low sensitivity to a relatively low AL dose. The present study was undertaken to examine whether resistance was mediated by differences in either systemic or beta-cell antioxidant defense status. Superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPX) activities were determined in tissues of AL untreated ALR/Lt and ALS/Lt male mice at 7 weeks of age. Specific activities of pancreatic SOD1, GR, and GPX were significantly increased in ALR/Lt mice compared with ALS/Lt mice. ALR/Lt mice further exhibited higher levels of glutathione in plasma, blood, pancreas, and liver combined with lower constitutive lipid peroxides in serum, liver, and pancreas. These results support the hypothesis that the selection process leading to the development of an AL-resistant mouse strain entailed accumulation of a gene or genes contributing to upregulated antioxidant status. PMID- 10468222 TI - Measurement of oxidatively induced base lesions in liver from Wistar rats of different ages. AB - Mitochondrial and nuclear DNA were isolated from the livers of young (6-7 month) and old (23-24 month) Wistar rats and the levels of 10 different oxidatively induced lesions were analyzed by gas chromatography/mass spectrometry. This is the first study to measure several different oxidatively induced base lesions in both mitochondrial and nuclear DNA as a function of age. No significant age effects were observed for any lesion. Furthermore, contrary to expectations, we did not observe elevated levels of oxidatively induced base lesions in mitochondrial DNA. This contrasts with 50-fold differences reported for several lesions between mitochondrial and nuclear DNA from porcine liver (Zastawny et al., Free Radic. Biol. Med. 24:722-725, 1998). The fact that different lesion levels are observed even when similar techniques are employed emphasizes that the role of oxidative mitochondrial DNA damage and its repair in aging must continue to be the subject of intense investigation. Questions concerning endogenous levels of damage should be revisited as existing methods are improved and new methods become available. PMID- 10468223 TI - Plasma levels of 8-epiPGF2alpha, an in vivo marker of oxidative stress, are not affected by aging or Alzheimer's disease. AB - Free radicals are likely involved in the aging process and there is a growing body of evidence that free radical damage to cellular function is associated with a number of age-related diseases such as atherosclerosis, cancer, and neurologic disorders. The present study was designed to evaluate in a healthy population the evolution with age of 8-epiPGF2alpha plasma levels, a recently proposed marker of in vivo lipid peroxidation. Moreover we investigated this marker of oxidative stress in patients with Alzheimer's disease (AD), an age-related neurodegenerative disorder in the development of which free radicals have been involved. Our results show that in the healthy population studied, despite decreased antioxidant defenses with increasing age as monitored by antioxidant capacity measurement, plasma 8-epiPGF2alpha levels were not correlated with age. Moreover, we have demonstrated that AD patients presented no modification of plasma 8-epiPGF2alpha level and no major alteration of the antioxidant status. In conclusion, the measurement of plasma 8-epiPGF2alpha did not allow us to detect alterations in oxidative stress with aging or in AD. PMID- 10468224 TI - Humic acid-mediated oxidative damages to human erythrocytes: a possible mechanism leading to anemia in Blackfoot disease. AB - Humic acid (HA) has been proposed as a factor that causes Blackfoot disease, an endemic peripheral vascular disease prevailing in the southwest coast of Taiwan. However, the relationship between HA and anemia associated with Blackfoot disease remains unclear. In this study, we showed that HA imposed damages on human red blood cells (RBCs), which were manifested as reduction in deformability of RBCs and hemolysis. At concentrations ranging from 10 to 100 microg/ml, HA caused lipid peroxidation in a dose-dependent manner. Such changes were accompanied by a depletion of glutathione and a reduction in activities of the antioxidant enzymes including catalase, superoxide dismutase, and glucose-6-phosphate dehydrogenase. These results indicate that HA initiates oxidative stress on RBCs and results in their dysfunction. Consistent with our previous findings, the present study supports the notion that HA plays an important role in the pathogenesis of Blackfoot disease. PMID- 10468225 TI - Elevation of hydrogen peroxide after spinal cord injury detected by using the Fenton reaction. AB - To reveal whether reactive oxygen species (ROS) play a role after spinal cord injury, we developed a unique method for assaying hydrogen peroxide (H2O2) and determined the time course of its concentration changes following impact injury to the rat spinal cord. Microdialysis was used to sample H2O2 in the extracellular space and the dialysates were collected into a vial containing salicylate and ferrous chloride (FeCl2). H2O2 collected in the vial was converted to hydroxyl radicals (*OH) by FeCl2 catalysis. 2,3- and 2,5-dihydroxybenzoic acid produced by reaction of *OH with salicylate in the collecting vial were measured by HPLC and calibrated to H2O2 concentrations. The postinjury levels of H2O2 were significantly increased (p = 0.02) for over 11 h. FeCl2 administered through the dialysis fiber catalyzes H2O2 conversion in the cord to *OH. This *OH does not reach the collecting vial due to its extremely short lifetime (nanoseconds). The reduced H2O2 levels in the vials validate the measurement of H2O2. The relatively long-lasting formation of H2O2 and superoxide reported herein and previously suggests that ROS may be important in secondary spinal cord damage and that removal of ROS may be a realistic treatment strategy for reducing injury caused by free radicals. PMID- 10468226 TI - Shear stress induction of C-type natriuretic peptide (CNP) in endothelial cells is independent of NO autocrine signaling. AB - C-type natriuretic peptide (CNP) is secreted by endothelial cells and has vasodilatory and antiproliferative activity against smooth muscle cells. Using defined laminar shear stress exposures of cultured bovine aortic endothelial cells, we investigated the regulation of CNP gene by PhosphorImaging the ratio of CNP mRNA to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA. A 6 h exposure to arterial shear stress of 25 dyn/cm2 caused a marked elevation (10.5 +/- 6.2 fold: n=10, p<0.001) of CNP/GAPDH mRNA ratio compared to stationary controls. Arterial shear stress was 2.6 times more potent than a venous level of shear stress of 4 dyn/cm2 in elevating the CNP/GAPDH mRNA ratio. After 6 h, CNP secretion by shear stressed BAEC was elevated over stationary controls by 3.1 fold (n=5, p<0.001) to a level of 34 +/- 7.5 pg/cm2 BAEC. Shear stress elevated CNP mRNA in the presence of L-NAME (400 microM) indicating that autocrine signaling through shear-induced NO production or guanylate cyclase activation was not involved. Similarly, the tyrosine kinase inhibitor genistein (10 microM), which can also block shear-induced NO production, had no effect on CNP mRNA induction by shear stress in BAEC. The intracellular calcium chelator BAPTA/AM (5 microM) attenuated the shear stress-induced CNP mRNA expression by 71%. Interestingly, dexamethasone (1 microM) potentiated by 2-fold the shear stress enhancement of CNP mRNA. Shear stress was a more potent inducer of CNP than either phorbol myristrate acetate or lipopolysaccharide. Hemodynamic shear stress may be an important physiological regulator of CNP expression with consequent effects on vasodilation and regulation of intimal hyperplasia. PMID- 10468227 TI - Electrical and morphological changes of human erythrocytes under high hydrostatic pressure followed by dielectric spectroscopy. AB - The electrical and morphological properties of human erythrocytes under high hydrostatic pressure up to 500 MPa have been studied by dielectric spectroscopy. The pressure-induced changes in the dielectric behavior of erythrocyte suspensions indicate that hydrostatic pressure causes the change in cell shape from discoidal to spherical and hemolysis at 200-300 MPa, the formation of buds and spicular processes followed by vesiculation at 300-400 MPa, and the increase in the membrane capacitance at 400-500 MPa. PMID- 10468228 TI - Computational simulation of platelet deposition and activation: I. Model development and properties. AB - To better understand the mechanisms leading to the formation and growth of mural thrombi on biomaterials, we have developed a two-dimensional computational model of platelet deposition and activation in flowing blood. The basic formulation is derived from prior work by others, with additional levels of complexity added where appropriate. It is comprised of a series of convection-diffusion-reaction equations which simulate platelet-surface and platelet-platelet adhesion, platelet activation by a weighted linear combination of agonist concentrations, agonist release and synthesis by activated platelets, platelet-phospholipid dependent thrombin generation, and thrombin inhibition by heparin. The model requires estimation of four parameters to fit it to experimental data: shear dependent platelet diffusivity and resting and activated platelet-surface and platelet-platelet reaction rate constants. The model is formulated to simulate a wide range of biomaterials and complex flows. In this article we present the basic model and its properties; in Part II (Sorensen et al., Ann. Biomed. Eng. 27:449-458, 1999) we apply the model to experimental results for platelet deposition onto collagen. PMID- 10468229 TI - Computational simulation of platelet deposition and activation: II. Results for Poiseuille flow over collagen. AB - We have previously described the development of a two-dimensional computational model of platelet deposition onto biomaterials from flowing blood (Sorensen et al., Ann. Biomed. Eng. 27:436-448, 1999). The model requires estimation of four parameters to fit it to experimental data: shear-dependent platelet diffusivity and three platelet-deposition-related reaction rate constants. These parameters are estimated for platelet deposition onto a collagen substrate for simple parallel-plate flow of whole blood in both the presence and absence of thrombin. One set of experimental results is used as a benchmark for model-fitting purposes. The "trained" model is then validated by applying it to additional test cases from the literature for parallel-plate Poiseuille flow over collagen at both higher and lower wall shear rates, and in the presence of various anticoagulants. The predicted values agree very well with the experimental results for the training cases, and good reproduction of deposition trends and magnitudes is obtained for the heparin, but not the citrate, validation cases. The model is formulated to be easily extended to synthetic biomaterials, as well as to more complex flows. PMID- 10468230 TI - Theoretical study of the effects of vascular smooth muscle contraction on strain and stress distributions in arteries. AB - To study the effects of smooth muscle contraction and relaxation on the strain and stress distribution in the vascular wall, a mathematical model was proposed. The artery was assumed to be a thick-walled orthotropic tube made of nonlinear, incompressible elastic material. Considering that the contraction of smooth muscle generates an active circumferential stress in the wall, a numerical study was performed using data available in the literature. The results obtained showed that smooth muscle contraction affects the residual strains which exist in a ring segment cut out from the artery and exposed to no external load. When the ring specimen is cut radially, it springs open with an opening angle. The predicted monotonic increase of the opening angle with increasing muscular tone was in agreement with recent experimental results reported in the literature. It was shown that basal muscular tone, which exists under physiological conditions, reduces the strain gradient in the arterial wall and yields a near uniform stress distribution. During temporary changes in blood pressure, the increase in muscular tone induced by elevated pressure tends to restore the distribution of circumferential strain in the arterial wall, and to maintain the flow-induced wall shear stress to normal level. PMID- 10468231 TI - In vivo three-dimensional surface geometry of abdominal aortic aneurysms. AB - Abdominal aortic aneurysm (AAA) is a local, progressive dilation of the distal aorta that risks rupture until treated. Using the law of Laplace, in vivo assessment of AAA surface geometry could identify regions of high wall tensions as well as provide critical dimensional and shape data for customized endoluminal stent grafts. In this study, six patients with AAA underwent spiral computed tomography imaging and the inner wall of each AAA was identified, digitized, and reconstructed. A biquadric surface patch technique was used to compute the local principal curvatures, which required no assumptions regarding axisymmetry or other shape characteristics of the AAA surface. The spatial distribution of AAA principal curvatures demonstrated substantial axial asymmetry, and included adjacent elliptical and hyperbolic regions. To determine how much the curvature spatial distributions were dependent on tortuosity versus bulging, the effects of AAA tortuosity were removed from the three-dimensional (3D) reconstructions by aligning the centroids of each digitized contour to the z axis. The spatial distribution of principal curvatures of the modified 3D reconstructions were found to be largely axisymmetric, suggesting that much of the surface geometric asymmetry is due to AAA bending. On average, AAA surface area increased by 56% and abdominal aortic length increased by 27% over those for the normal aorta. Our results indicate that AAA surface geometry is highly complex and cannot be simulated by simple axisymmetric models, and suggests an equally complex wall stress distribution. PMID- 10468232 TI - Pulse pressure method and the area method for the estimation of total arterial compliance in dogs: sensitivity to wave reflection intensity. AB - We estimated total arterial compliance (C) in eight anesthetized mongrel dogs with (i) the area method (AM), (ii) the pulse pressure method (PPM), and (iii) the stroke volume-to-pulse pressure ratio (SV/PP). Average compliance was C(AM)=1.1+/-0.73 ml mm Hg(-1) using AM; C(PPM)=0.60+/-0.31 ml mm Hg(-1) using PPM and C(SV/PP)=0.87+/-0.49 ml mm Hg(-1) using SV/PP. Mean aortic pressure was 64+/ 23 mm Hg. The overall agreement between C(AM) and C(PPM) was relatively poor (C(AM)=0.15+/-1.61 C(PPM); r2=0.48), with a consistent overestimation of the area method with respect to the pulse pressure method. There was a significant correlation (r=-0.78) between the relative difference between PPM and AM, and the modulus of the first harmonic of the wave reflection coefficient [gamma] which was low in our dog population (0.37+/-0.18). SV/PP overestimated PPM, but both methods were highly correlated (C(SV/PP)=0.06+/-1.60C(PPM); r2=0.97). C(SV/PP) and C(AM) were similar only for [gamma]>0.4. The effect of isolated changes of [gamma] on PPM, AM, and SV/PP was studied using the linear wave separation technique. The area method appeared very sensitive to the wave reflection intensity. For low reflection coefficients, the diastolic wave profile was flattened and compliance was overestimated. PPM and SV/PP were relatively independent of [gamma] and remained even applicable for [gamma]=0. We believe that the pulse pressure method is the most consistent method for the estimation of total arterial compliance in hemodynamic conditions characterized by a low wave reflection intensity. PMID- 10468233 TI - Fast estimation of arterial vascular parameters for transient and steady beats with application to hemodynamic state under variant gravitational conditions. AB - Numerous parameter estimation techniques exist for characterizing the arterial system using electrical circuit analogs. These techniques are often limited by requiring steady-state beat conditions and can be computationally expensive. Therefore, a new method was developed to estimate arterial parameters during steady and transient beat conditions. A four-element electrical analog circuit was used to model the arterial system. The input impedance equations for this model were derived and reduced to their real and imaginary components. Next, the physiological input impedance was calculated by computing fast Fourier transforms of physiological aortic pressure (AoP) and aortic flow. The approach was to reduce the error between the calculated model impedance and the physiological arterial impedance using a Jacobian matrix technique which iteratively adjusted arterial parameter values. This technique also included algorithms for estimating physiological arterial parameters for nonsteady physiological AoP beats. The method was insensitive to initial parameter estimates and to small errors in the physiological impedance coefficients. When the estimation technique was applied to in vivo data containing steady and transient beats it reliably estimated Windkessel arterial parameters under a wide range of physiological conditions. Further, this method appears to be more computationally efficient compared to time-domain approaches. PMID- 10468234 TI - Direct inference of the spectra of pericardial potentials using the boundary element method. AB - New methods, based on Tikhonov regularization, were developed to infer the magnitude and phase of pericardial potentials directly. These methods were tested in an adult-male torso model using measured human epicardial potentials. With 1% noise added to body-surface potentials, regularization with an optimal parameter at each frequency from 1 to 100 Hz gave an average relative error (RE) in inferred spectral magnitudes of 0.44. Regularization with the composite-residual smoothing-operator (CRESO) parameter increased the RE slightly to 0.47. With 10% additive noise, 10 mm overestimation of heart radius, and a 10 mm error in heart position, the average CRESO parameter from 1 to 100 Hz gave an average RE of 0.71. Performance was frequency dependent. The smallest REs occurred at low frequencies. With 1% noise, optimal regularization gave average REs of 0.20, 0.40, and 0.53 in the 1-15, 15-46, and 46-100 Hz bands, respectively. Direct inference of spectral magnitudes was more accurate than Fourier transformation of inferred time-domain waveforms. Results suggest that when heart size and location are not known, minimum REs in spectral estimates are found using an overestimated heart size and a regularization parameter which is the average value over the frequency band of interest. PMID- 10468235 TI - Model for intravital microscopic evaluation of the effects of arterial occlusion caused ischemia in bone. AB - An in vivo model has been developed for chronic observation of the effects of ischemia on cortical bone remodeling and perfused vascularity. Diaphragm occluders were implanted around the right common iliac artery of four rabbits and inflated to produce 10 h of ischemia to the limb. Microcirculation was monitored with intravital microscopy of injected fluorescent microspheres and FITC-Dextran 70 through a bone window, the tibial bone chamber implant (BCI). Bone resorption and apposition in the BCI were indicated with mineralization dyes. Between 2 and 12 h following release of the occluder, secondary ischemia/no-reflow and other evidence of reperfusion injury were observed. Vessel damage was suggested by abnormally high leakage of FITC-D70 from the few vessels perfused during secondary ischemia. In the weeks following occluder release perfused vasculature increased beyond pre-occlusion levels. Net bone resorption reached a maximum when vascularity passed normal levels. In order to further validate the arterial occlusion model for osteonecrosis, techniques for (1) confirming bone death and (2) detecting increased leukocyte adherence to endothelial cells were added. The dead cell stain Ethidium homodimer-1 was used to tag dead osteocytes immediately after occlusion and produced a measure designated "osteonecrosis index." To detect leukocytes adhering to vessel walls, carboxyfluorescein diacetate, succinimidyl ester was injected at occluder release. An increase in the number of adherent leukocytes was detected. PMID- 10468236 TI - Dependence of intertrabecular permeability on flow direction and anatomic site. AB - The structure-function relationships for the permeability of trabecular bone may have relevance for tissue engineering, total joint replacements, and whole bone mechanics. To investigate such relationships, we used a constant flow rate permeameter to determine the intrinsic permeability of trabecular bone specimens, oriented longitudinally or transversely to the principal trabecular orientation, from the human vertebral body (n=20), human proximal femur (n=12), and bovine proximal tibia (n=24). Overall, the intertrabecular permeability ranged from 2.68 x 10(-11) to 2.00 x 10(-8) m2. Significant negative nonlinear relations between intertrabecular permeability and volume fraction were found for each group except the longitudinal bovine proximal tibial specimens (r2=0.34-0.80). The average permeability ratio, a measure of the anisotropy, was 2.05, 6.60, and 23.3 for the human vertebral body, bovine tibia, and human femur, respectively. The permeability depended strongly on flow direction relative to the principal trabecular orientation (p<0.0001) and anatomic site (p <0.0001). In addition to providing a comprehensive description of intertrabecular permeability as a function of anatomic site and flow direction, these data provide substantial insight into the underlying structure-function relationships. PMID- 10468237 TI - Inference of blood glucose concentrations from subcutaneous glucose concentrations: applications to glucose biosensors. AB - An approach for inference of blood glucose concentrations in real time is considered. First, a model that predicts the subcutaneous glucose concentration from the blood glucose concentration is presented. The model includes diffusive transport from the blood to the subcutaneous tissue and reactive-like cellular uptake of glucose. Next, the Phillips-Tikhonov regularization method is considered to solve the real-time input estimation problem that determines the blood glucose concentration given the subcutaneous glucose concentration. The inverse problem is regularized by imposing a smoothing condition to obtain a stable solution. Three different penalization functionals were considered in evaluating the regularization method using a synthetic function that approximates the subcutaneous glucose response to an oral glucose tolerance test in a human subject. Various levels of either white noise or time-correlated noise were superimposed onto the synthetic response to evaluate the sensitivity of the inverse to measurement error. For inversion assuming only diffusive transport, the optimal time interval of integration of previous subcutaneous measurements was found to be about 1.5/alpha, where alpha(-1) is the dominant time constant for the exchange of glucose between the blood and subcutaneous tissue. The optimal sampling rate was found to be 54alpha. Linear regularizations based on minimization of first or second derivatives of the blood glucose concentration were found to be satisfactory, each yielding a minimum error that was about 50% greater than the measurement error. Including nonlinear, reactive-like uptake of glucose was found to decrease the error magnification factor slightly. Both the model and the inverse method relating blood and subcutaneous glucose concentrations are successfully applied to experimental measurements using glucose biosensors reported by Schmidtke et al. (Proc. Natl. Acad. Sci. USA 95:294-299, 1998). PMID- 10468238 TI - Robust nonlinear autoregressive moving average model parameter estimation using stochastic recurrent artificial neural networks. AB - In this study, we introduce a new approach for estimating linear and nonlinear stochastic autoregressive moving average (ARMA) model parameters, given a corrupt signal, using artificial recurrent neural networks. This new approach is a two step approach in which the parameters of the deterministic part of the stochastic ARMA model are first estimated via a three-layer artificial neural network (deterministic estimation step) and then reestimated using the prediction error as one of the inputs to the artificial neural networks in an iterative algorithm (stochastic estimation step). The prediction error is obtained by subtracting the corrupt signal of the estimated ARMA model obtained via the deterministic estimation step from the system output response. We present computer simulation examples to show the efficacy of the proposed stochastic recurrent neural network approach in obtaining accurate model predictions. Furthermore, we compare the performance of the new approach to that of the deterministic recurrent neural network approach. Using this simple two-step procedure, we obtain more robust model predictions than with the deterministic recurrent neural network approach despite the presence of significant amounts of either dynamic or measurement noise in the output signal. The comparison between the deterministic and stochastic recurrent neural network approaches is furthered by applying both approaches to experimentally obtained renal blood pressure and flow signals. PMID- 10468239 TI - Parametric and nonparametric nonlinear system identification of lung tissue strip mechanics. AB - Lung parenchyma is a soft biological material composed of many interacting elements such as the interstitial cells, extracellular collagen-elastin fiber network, and proteoglycan ground substance. The mechanical behavior of this delicate structure is complex showing several mild but distinct types of nonlinearities and a fractal-like long memory stress relaxation characterized by a power-law function. To characterize tissue nonlinearity in the presence of such long memory, we investigated the robustness and predictive ability of several nonlinear system identification techniques on stress-strain data obtained from lung tissue strips with various input wave forms. We found that in general, for a mildly nonlinear system with long memory, a nonparametric nonlinear system identification in the frequency domain is preferred over time-domain techniques. More importantly, if a suitable parametric nonlinear model is available that captures the long memory of the system with only a few parameters, high predictive ability with substantially increased robustness can be achieved. The results provide evidence that the first-order kernel of the stress-strain relationship is consistent with a fractal-type long memory stress relaxation and the nonlinearity can be described as a Wiener-type nonlinear structure for displacements mimicking tidal breathing. PMID- 10468240 TI - Errors caused by combination of Di-4 ANEPPS and Fluo3/4 for simultaneous measurements of transmembrane potentials and intracellular calcium. AB - Intracellular calcium concentration and transmembrane potentials are two important measurements used to study the mechanisms of cardiac activity. Fluorescent dyes have been used to measure these separately but not simultaneously in cardiac tissue. Fluo-3 and Fluo-4 (a recently improved version of Fluo-3) have been used to measure changes in intracellular calcium concentration and Di-4 ANEPPS has been used to measure transmembrane potentials. This paper addresses the feasibility of using these fluorescent dyes together in order to measure transmembrane potentials and intracellular calcium concentration simultaneously. For the dyes to be used simultaneously, their respective fluorescence spectra must be sufficiently separated in wavelength in order to allow them to be separated by optical filters or spectrographs. An apparatus was constructed to measure the dyes' spectra in a fluorescence imaging chamber as well as in an isolated perfused rabbit heart. The measured spectra were mathematically modeled in order to assess the spectral overlap error under different conditions. Error graphs were constructed which may help researchers select optical filters and dye concentrations that will result in an acceptable error. PMID- 10468241 TI - Estimation of the shear stress on the surface of an aortic valve leaflet. AB - The limited durability of xenograft heart valves and the limited supply of allografts have sparked interest in tissue engineered replacement valves. A bioreactor for tissue engineered valves must operate at conditions that optimize the biosynthetic abilities of seeded cells while promoting their adherence to the leaflet matrix. An important parameter is shear stress, which is known to influence cellular behavior and may thus be crucial in bioreactor optimization. Therefore, an accurate estimate of the shear stress on the leaflet surface would not only improve our understanding of the mechanical environment of aortic valve leaflets, but it would also aid in bioreactor design. To estimate the shear stress on the leaflet surface, two-component laser-Doppler velocimetry measurements have been conducted inside a transparent polyurethane valve with a trileaflet structure similar to the native aortic valve. Steady flow rates of 7.5, 15.0, and 22.5 L/min were examined to cover the complete range possible during the cardiac cycle. The laminar shear stresses were calculated by linear regression of four axial velocity measurements near the surface of the leaflet. The maximum shear stress recorded was 79 dyne/cm2, in agreement with boundary layer theory and previous experimental and computational studies. This study has provided a range of shear stresses to be explored in bioreactor design and has defined a maximum shear stress at which cells must remain adherent upon a tissue engineered construct. PMID- 10468242 TI - Maintenance of homeostasis in coagulation during cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) represents a massive stimulus that overwhelms the body's homeostatic ability to buffer. Contact activation, the extrinsic cascade, and ongoing cellular interactions all must be considered to understand the coagulation dysfunction that occurs in CPB. Various aspects and the specific effects of these factors as well as their interrelationships are still poorly understood. Since we are dealing with a complex system, the way we approach our investigations must become more rather than less complex. Simply measuring peripheral blood levels of various proteins throughout CPB levels does not fit the important physiologic processes governing homeostasis, as production, clearance, redistribution, and dilution are all constantly at play. A recent study of prothrombin fragment F1.2 levels during CPB is provided as an illustration. Our increasing understanding of aprotinin's mechanisms of action has furthered our basic understanding of coagulation dysfunction in CPB. Further progress requires new thinking, more research, and increasingly complex approaches to the understanding of what drives homeostasis. PMID- 10468243 TI - Hemostatic agents and their safety. AB - The pharmacologic management of hemostasis in patients undergoing cardiopulmonary bypass may be accompanied by adverse responses. Evaluating the safety profile of hemostatic agents (eg, lysine analogs, aprotinin, protamine, or even donor blood) should be done objectively. Subsequent to early anecdotal reports, the safety profile of aprotinin, a broad-spectrum serine protease inhibitor, has been thoroughly evaluated in multiple double-blind, placebo-controlled, multicenter studies. Although associated with decreased fibrinolysis, aprotinin has not been associated with an increased risk of post-cardiopulmonary bypass myocardial infarction, graft closure, stroke, or increased risk of renal dysfunction from US studies. As with any polypeptide, there is a risk of anaphylaxis, which is influenced not only by prior exposure but also by time since prior exposure. In a similar fashion, after early anecdotal reports, evaluations involving large numbers of patients have helped define adverse reactions to protamine. Adverse reactions to blood products also must be considered in any safety comparisons involving hemostatic agents. PMID- 10468244 TI - Etiology and incidence of brain dysfunction after cardiac surgery. AB - The frequency and severity of central nervous system complications in patients undergoing cardiopulmonary bypass (CPB) may be greater than previously thought, particularly in the older population. The risks of embolic neurologic complications and stroke in the population older than 70 years from a severely atherosclerotic ascending aorta are well documented. Moreover, while the majority of CPB patients do not experience perioperative stroke, a high incidence of more subtle central nervous system dysfunction has been demonstrated to persist for up to 1 year after surgery. This report reviews the incidence and severity of cerebral injury during CPB and the effects of both age and the severely atherosclerotic ascending aorta on adverse neurologic outcomes. It discusses perioperative diagnostic methods, including transesophageal echocardiography, periaortic echocardiography, transcranial Doppler, and retinal fluorescein angiography, and the benefit of pH management. Ischemic brain injury resulting from activation of injury-related enzymes as part of the systemic inflammatory response is briefly reviewed. Age has been shown to be the strongest predictor of neurologic sequelae in patients undergoing CPB. The risk of embolic complications in the brain also increases in proportion to the degree of atherosclerosis in the ascending aorta, which is age-related. Transesophageal echocardiography has been found to be only partly useful in diagnosing these lesions or in guiding surgical manipulations in comparison with epiaortic imaging, which is more discreet. Transcranial Doppler and retinal fluorescein angiography have provided further evidence of microemboli during surgical manipulations. In a 316-patient prospective study, we found no differences in outcome between pH-stat and alpha stat strategies during moderate hypothermic CPB, except in patients who were on bypass for more than 90 minutes. Approximately 90% of these had a significant reduction in cognitive impairment with the alpha-stat method. Aprotinin, a serine protease inhibitor, has been found in two separate, randomized, placebo controlled trials to significantly lower incidences of perioperative stroke. Further study to develop therapeutic and preemptive strategies for prevention of brain injury is required, especially in the elderly. Aprotinin and other modalities aimed at suppressing the inflammatory response to CPB may offer hope because they act to suppress injury-provoking enzymes and leukocyte activation that are, in part, responsible for organ system dysfunction following CPB. PMID- 10468245 TI - Anticoagulation and anticoagulation reversal with cardiac surgery involving cardiopulmonary bypass: an update. AB - Accelerated thrombin generation is central to the development of hemostatic abnormalities during cardiopulmonary bypass (CPB) that are associated with both thromboembolic complications and serious, abnormal bleeding. Thrombin not only converts fibrinogen to fibrin, but also activates platelets and coagulation factors V, VIII, and XI and causes release of von Willebrand factor from vascular endothelium. Thrombin can also downregulate the hemostatic system by inducing formation of platelet inhibitory agents, such as nitric oxide and prostacyclin, and release of tissue plasminogen activator, facilitating activation of protein C, and releasing tissue factor pathway inhibitor. Excessive thrombin activity may also result in substantial consumption of platelets, fibrinogen, and labile coagulation factors and abnormal bleeding. Elevated tissue plasminogen activator levels secondary to activation of the contact system and surgery catalyze the formation of plasmin, which also consumes or internalizes platelet glycoprotein receptors and coagulation factors V, VIII, and fibrinogen. Heparin can reduce the generation of and mediate neutralization of excessive and CPB-associated thrombin activity. Heparin anticoagulation is commonly monitored with the activated clotting time (ACT). However, the ACT may be prolonged by factors other than heparin during CPB, such as hemodilution and hypothermia, and therefore may not accurately reflect the extent of anticoagulation by heparin. Aprotinin, a nonspecific serine protease inhibitor used with CPB, can also prolong celite based ACT values, rendering it less reliable for monitoring heparin anticoagulation. Therefore, several alternative anticoagulation strategies have been recommended when aprotinin is used, such as a higher celite ACT trigger (>750 seconds), monitoring of whole blood heparin concentrations (eg, >2.7 U/mL), or administration of heparin based on a CPB duration-dependent, fixed-dose regimen. Administration of heparin doses higher than those generally recommended, as guided by predetermined, patient-specific whole blood heparin concentration measurements during bypass, can reduce excessive thrombin-mediated consumption of platelets and coagulation factors as well as post-CPB blood loss and blood component transfusions. New modalities of improving suppression of excess thrombin generation during CPB include use of heparin-bonded CPB circuits, heparin cofactor II or related analogs, supplemental antithrombin III, direct thrombin inhibitors (eg, hirudin, argatroban), and inhibitors of the contact and tissue factor pathways. The safety and efficacy of these approaches remains to be established by additional, appropriately powered, prospective studies. PMID- 10468246 TI - Microvascular responses to cardiopulmonary bypass. AB - Cardiopulmonary bypass can result in proinflammatory and procoagulant changes that can contribute to morbidity and mortality in heart surgery patients. These responses, many of which are mediated by activation of endothelial cells, normally serve to repair damaged tissue or as defenses against infection. Once activated in the setting of surgery and trauma, these responses may cause unwarranted tissue destruction if they occur inappropriately or too diffusely. The proinflammatory response results in the release of cytokines and subsequent localization of neutrophils, which can disrupt the endothelial barrier and damage underlying tissue. The procoagulant response is characterized by the transcriptional activation of tissue factor, subsequent thrombin generation with subsequent microvascular thrombosis. Techniques to inhibit endothelial cell activation while attempting to preserve the body's anti-infectious and repair mechanisms are being investigated. These include hypothermia, blockade of adhesion molecules, blocking of chemotactic factors such as interleukin-8, and prevention of transcriptional activation by inhibiting the action of nuclear factor kappa-B, which activates genes involved in this process. PMID- 10468247 TI - Aprotinin has direct platelet protective properties: fact or fiction? PMID- 10468248 TI - Platelet function during cardiac surgery and cardiopulmonary bypass with low-dose aprotinin. AB - OBJECTIVE: To determine whether two low-dose regimens of aprotinin influence platelet function. DESIGN: Prospective, randomized, single-blinded trial. SETTING: University teaching hospital performing 600 cardiac operations per year. PARTICIPANTS: Fifty-nine patients scheduled for cardiac surgery undergoing cardiopulmonary bypass (CPB) of expected duration of 60 minutes or more. INTERVENTIONS: Patients were randomized into three groups. Group C (control) included 21 patients who did not receive aprotinin. In group A2, 17 patients received 14,286 kallikrein inhibitor units (KIU)/kg (2 mg/kg) of aprotinin before surgery, followed by a continuous infusion of 7,143 KIU/kg/h (1 mg/kg/h) until the end of surgery. In group A4, 19 patients received 28,572 KIU/kg (4 mg/kg) of aprotinin before surgery, followed by the same infusion. MEASUREMENTS AND MAIN RESULTS: Postoperative bleeding and transfusion requirements were significantly less in group A4. Changes in platelet number and function were similar in the three groups. Platelet aggregation was assessed in four periods: before CPB (T1), post-CPB (T2), and 2 hours (T3) and 4 hours (T4) after CPB. Platelet aggregation induced by adenosine diphosphate, 1 and 2 micromol/L; ristocetin, 1 mg/mL; and arachadonic acid (AA), 1.4 mmol/L, decreased at T2 (p < 0.001) in all groups, and for the ristocetin and AA groups, remained at less than baseline values at T3 and T4. In five patients from each group, platelet receptors for glycoprotein IIb IIIa (GPIIb-IIIa) and expression of platelet activation markers, guanosine monophosphate 140 (GMP-140) and lysosomal protein, were measured by flow cytometry before and after CPB. Modifications in the expression of GPIIb-IIIa were always modest and without statistical significance. Platelet activation markers, GMP-140 or lysosomal protein, nearly doubled from baseline to post-CPB only in the A4 group, whereas they remained stable in both other groups (statistically not significant). CONCLUSION: The two regimens of aprotinin, both considered low dosage, did not exert a protective effect on platelet function. Neither dose produced changes in platelet GPIIb-IIIa or platelet activation markers. However, bleeding and transfusion needs were decreased. PMID- 10468249 TI - Platelet protection in coronary artery surgery: benefits of heparin-coated circuits and high-dose aprotinin therapy. AB - OBJECTIVE: To examine the extent of platelet activation during extracorporeal circulation by using the combination of heparin-coated oxygenation systems and high-dose aprotinin therapy, and to examine the affinity and thereby the protective capacity of aprotinin to the glycoprotein (GP) receptors of the platelet membrane. DESIGN: Experimental in vitro study. SETTING: Research laboratory of a university hospital. PARTICIPANTS: Thirty-two volunteers (blood donors). MEASUREMENTS AND MAIN RESULTS: Thirty-two oxygenation circuits of the same construction series (16 heparin-coated and 16 noncoated) were investigated in a closed system of a heart-lung machine model with fresh human whole blood. In each of these two groups, eight circuits with and eight without a high-dose aprotinin application (250 kallikrein inhibitory units [KIU]/mL) were investigated. In all four groups, the number of platelets declined continuously during the 90-minute recirculation period. Group I (no heparin coating, no aprotinin) showed the greatest reduction; group IV (heparin coating, aprotinin) had a significantly smaller decrease in platelet number (p < 0.01). Platelet factor 4 (PF-4) levels, released from the alpha-granule, were in inverse proportion to the platelet loss. After 90 minutes of recirculation, the PF-4 values increased to 615.8% +/- 559.5% and 237.2% +/- 179.0% of the initial value for groups I and IV, respectively (p < 0.01). Affinity chromatography and immunoblotting techniques were used to evaluate the affinity of aprotinin for the GP receptors of the platelet membrane. The affinity appeared in the following order: GPIIb < GPIIIa < GPIb. CONCLUSION: Heparin-coated oxygenation systems and additional aprotinin caused significantly less platelet damage in an in vitro cardiopulmonary bypass model. Chromatographic and immunologic methods could prove aprotinin's affinity for the platelet receptor proteins GPIb and GPIIb-IIIa and therefore its probable role in diminishing the triggering of the platelet activation cascade. PMID- 10468250 TI - Sonoclot coagulation analysis: a study of test variability. AB - OBJECTIVE: To test the reproducibility of Sonoclot coagulation analysis (SCA; Sienco Inc, Morrison, CO). The authors wished to determine if the mix/release of the preloaded celite activator in standard Sono-cuvettes could be responsible for the high variation coefficients for SCA parameters with citrated whole blood and if citrated whole blood is optimal for SCA. DESIGN: A prospective trial. SETTING: A large academic teaching medical center. PARTICIPANTS: Eight healthy volunteers. INTERVENTIONS: Repeated blood sampling was performed through indwelling radial artery catheters. Seven different Sonoclot analyzers were used to test seven different types of analysis procedures in the volunteers, involving activators of different types and amounts and in different forms, and the use of citrated or native whole blood. MEASUREMENTS AND MAIN RESULTS: Two-way and one-way ANOVA, variance, variance analysis, and Tukey's test were used to evaluate differences in SCA methods and volunteer influence. A high variance, with SDs up to 200% of the median values of the SCA parameters with recalcified citrated blood and the standard Sono-cuvette, was confirmed. SCA with native blood and/or the use of other types of preloaded activators, ie, kaolin, significantly (p < 0.05) reduced this variance. Repeated SCAs further reduced the variance to 10% to 35% of the variance for a single analysis (standard cuvette and native blood). CONCLUSION: Improvement of the activator in the Sonocuvette, use of native whole blood, and repeated Sonoclot analyses reduced the previously reported high variability of this instrument. PMID- 10468251 TI - Coagulation tests during cardiopulmonary bypass correlate with blood loss in children undergoing cardiac surgery. AB - OBJECTIVES: To examine whether coagulation tests, sampled before and during cardiopulmonary bypass (CPB), are related to blood loss and blood product transfusion requirements, and to determine what test value(s) provide the best sensitivity and specificity for prediction of excessive hemorrhage. DESIGN: Prospective. SETTING: University-affiliated, pediatric medical center. PARTICIPANTS: Four hundred ninety-four children. INTERVENTIONS: Coagulation tests. MEASUREMENTS AND MAIN RESULTS: Demographic, coagulation test, blood loss, and transfusion data were noted in consecutive children undergoing cardiac surgery. Laboratory tests included hematocrit (Hct), prothrombin time, partial thromboplastin time (PTT), platelet count, fibrinogen concentration, and thromboelastography. Stepwise linear regression analysis indicated that platelet count during CPB was the variable most significantly associated with intraoperative blood loss (in milliliters per kilogram) and 12-hour chest tube output (in milliliters per kilogram). Other independent variables associated with blood loss were thromboelastography maximum amplitude (MA) during CPB, preoperative PTT, preoperative Hct, and preoperative thromboelastography angle and shear modulus values. Thromboelastography MA during CPB was the only variable associated with total products transfused (in milliliters per kilogram). Of all tests studied, platelet count during CPB (< or = 108,000/microL) provided the maximum sensitivity (83%) and specificity (58%) for prediction of excessive blood loss (receiver operating characteristic analysis). Blood loss was inversely related to patient age; neonates received the most donor units (median, 8 units; range, 6 to 10 units). CONCLUSIONS: During cardiac surgery, coagulation tests (including thromboelastography) drawn pre-CPB and during CPB are useful to identify children at risk for excessive bleeding. Platelet count during CPB was the variable most significantly associated with blood loss. PMID- 10468253 TI - Safety and efficacy of shed mediastinal blood transfusion after cardiac surgery: a multicenter observational study. Multicenter Study of Perioperative Ischemia Research Group. AB - OBJECTIVE: To examine the efficacy and safety of shed mediastinal blood (SMB) transfusion in preventing allogenic red blood cell (RBC) transfusion. DESIGN: An observational clinical study. SETTING: Twelve US academic medical centers. PARTICIPANTS: Six hundred seventeen patients undergoing elective primary coronary artery bypass grafting. INTERVENTIONS: Patients were administered SMB transfusion or not, according to institutional and individual practice, without random assignment. MEASUREMENTS AND RESULTS: The independent effect of SMB transfusion on postoperative RBC transfusion was examined by multivariable modeling. Potential complications of SMB transfusion, such as bleeding and infection, were examined. Three hundred twelve of the study patients (51%) received postoperative SMB transfusion (mean volume, 554 +/- 359 mL). Patients transfused with SMB had significantly lower volumes of RBC transfusion than those not receiving SMB (0.86 +/- 1.50 v 1.08 +/- 1.65 units; p < 0.05). However, multivariable analysis showed that SMB transfusion was not predictive of postoperative RBC transfusion. Demographic factors (older age, female sex), institution, and postoperative events (greater chest tube drainage, lower hemoglobin level on arrival to the intensive care unit, and use of inotropes) were significant predictors of RBC transfusion. The volume of chest tube drainage on the operative day (707 +/- 392 v 673 +/- 460 mL; p = 0.30), reoperation for hemorrhage (3.1% v2.5%; p = 0.68), and overall frequency of infection (5.8% v 6.6%; p = 0.81) were similar between patients receiving and not receiving SMB, respectively. However, in patients who did not receive allogenic RBC transfusion, there was a significantly greater frequency of wound infection in the SMB group (3.6% v0%; p = 0.02). CONCLUSION: These data suggest that SMB is ineffective as a blood conservation method and may be associated with a greater frequency of wound infection. PMID- 10468252 TI - Do anticoagulation with heparin and protamine reversal alter thrombogenicity of coated and noncoated pulmonary artery catheters? AB - OBJECTIVE: To clarify the role of anticoagulation with heparin and protamine reversal on the effectiveness of heparin bonding or coating of pulmonary artery catheters in preventing thrombus formation in the Rhesus monkey. DESIGN: A controlled, unblinded, open-labeled study. SETTING: A research laboratory at Tulane School of Medicine (New Orleans, LA). PARTICIPANTS: Twenty-four anesthetized Rhesus monkeys. INTERVENTIONS: The monkeys were assigned to one of four groups. In the first group (group A), non-heparin-coated catheters were inserted into a femoral vein through an incision, advanced proximally for a distance of 30 cm, and left in place for 1 hour. In the second group (group B), heparin-coated catheters were placed and evaluated in the same manner as in group A. In the third group (group C), the primates received 3 mg/kg of heparin intravenously (i.v.) before insertion of a non-heparin-coated pulmonary artery catheter. After the catheter had been in place for 1 hour, protamine, 3 mg/kg i.v., was administered, and the catheter was left in place for an additional hour. In the final group (group D), the primates received 3 mg/kg of heparin i.v. before insertion of a heparin-coated pulmonary artery catheter. After the catheter had been in place for 1 hour, protamine, 3 mg/kg i.v., was administered, and the catheter was left in place for an additional hour. In each group, the catheter was withdrawn with the balloon inflated to minimize any stripping of thrombus from the surface of the catheter. Thrombus was removed from the catheter through a femoral vein cutdown and weighed. Laboratory values were determined for each animal, and clot formation was evaluated in each group. After completion of the study, the animals were returned to the primate breeding colony. MEASUREMENTS AND MAIN RESULTS: There were no significant differences in hematocrit, prothrombin time, partial thromboplastin time, or platelet levels among the four groups; therefore, the data were pooled. Clots were observed on five of six catheters in both groups A and C; however, clot formation was seen in one of six catheters in group B and three of six catheters in group D. There was a statistically significant difference (p < 0.01) in mean clot weight in group A (265 +/- 68 mg; range, 0 to 447 mg) compared with 13 +/- 13 mg in group B (range, 0 to 80 mg). There was no significant difference in mean clot weight in group C (104 +/- 35 mg; range, 0 to 202 mg) compared with group D (24 +/- 16 mg, range, 0 to 98 mg). Additionally, in group C, the mean clot weight was significantly less than in group A. CONCLUSION: Anticoagulation of primates with heparin before catheter insertion imparts some protection to non-coated catheters, and protamine reversal of anticoagulation with heparin may partially negate the protective effect seen with heparin-coated pulmonary artery catheters. PMID- 10468254 TI - Minimally invasive direct coronary artery bypass grafting: changes in anesthetic management and surgical procedure. AB - OBJECTIVE: The authors hypothesized that changes in surgical procedures for minimally invasive direct coronary artery bypass grafting (MIDCAB) have led to changes in anesthetic management with a resultant decrease in the complexity of care. DESIGN: Retrospective observational study. SETTING: University teaching hospital. PARTICIPANTS: Review of the records of 60 patients who underwent MIDCAB surgery. MEASUREMENTS AND MAIN RESULTS: Data included preoperative demographics, perioperative anesthetic management, and postoperative cardiac and noncardiac issues and complications. Two groups were formed: in group I, a coronary stabilizer (CS) was not used, and in group II, it was. With the exception of a greater incidence of those with no preoperative comorbidities in group II (CS), there were no differences between the two groups with respect to demographics or preoperative variables. A surgical design called H-graft was used in a greater number of group II (CS) patients, whereas a direct anastomosis was performed in the majority of group I patients. Use of pharmacologically induced bradycardia/asystole has not been performed after the introduction of the CS. The use of central venous catheters (instead of pulmonary artery catheters) and single-lumen (v double-lumen) endotracheal tubes was greater in group II (CS) patients. Despite changes in intraoperative management, there was no significant change in the incidence of postoperative complications, intensive care unit stay, and hospital stay between groups I and II. New-onset atrial fibrillation was the most common postoperative complication (13 of 56 patients; 23%). Three of 24 patients (12.5%) who received intraoperative magnesium experienced atrial fibrillation compared with 10 of 32 patients (31%) who did not receive magnesium. CONCLUSIONS: The complexity of anesthetic technique has decreased since the onset of MIDCAB surgery. The decrease in complexity may be related to changes in surgical design and technology. PMID- 10468255 TI - Influence of fast-track anesthetic technique on cardiovascular infusions and weight gain. AB - OBJECTIVE: To evaluate whether cardiac surgical patients receiving conventional versus fast-track anesthetic management are statistically significantly different with regard to cardiovascular drug infusions, weight gain, cardiac and pulmonary morbidity, length of intubation, and length of stay. DESIGN: Retrospective, (partially) sequential, cohort design. SETTING: Surgical suite and intensive care unit (ICU) at a community hospital. PARTICIPANTS: Two hundred seven patients who presented for coronary artery bypass graft and/or cardiac valve replacement. INTERVENTIONS: None MEASUREMENTS AND MAIN RESULTS: Group comparisons of the seven individual cardiovascular drug infusions showed less frequent use in the fast track patients for lidocaine (9% v 28%; p = 0.00046) only. However, the fast track group received fewer combinations of cardiovascular drug infusions overall for the first 24-hour postoperative period (p < 0.0005). Hourly comparisons of inotropes showed significantly fewer combinations of dobutamine, norepinephrine, and epinephrine for the first postoperative hour and for postoperative hours 7 through 12 (p < 0.01 for each hour). Fast-track patients had less postoperative weight gain for days 1 through 4 (p < 0.01 for each day), shorter length of ICU stay (p < 0.00005), and shorter total length of postoperative hospital stay (p = 0.0004). No differences were found with respect to myocardial infarction, death, pulmonary complications, rate of reintubation, or length of hospital stay once discharged from the ICU. CONCLUSIONS: Fast-track anesthetic management may be associated with decreased need for inotropic and antiarrhythmic drug infusions and decreased weight gain. PMID- 10468256 TI - Anesthetic management of patients undergoing coronary artery bypass grafting with the use of an axial flow pump and a short-acting beta-blocker. AB - OBJECTIVES: To describe the clinical protocol regarding monitoring, pharmacologic interventions, and postoperative care during and after coronary artery bypass grafting (CABG) on the beating heart with an axial flow pump and a short-acting beta-blocker. DESIGN: A retrospective study. SETTING: A university hospital. PARTICIPANTS: Seventeen patients scheduled for elective CABG. INTERVENTIONS: Invasive monitoring was performed with either a standard pulmonary artery catheter (PAC) or a surgically placed PAC. An axial flow pump was inserted through a graft sutured to the ascending aorta. A short-acting beta-blocker was administered to decrease the motion of the heart and make conditions for CABG adequate and safe. MEASUREMENTS AND MAIN RESULTS: Compared with baseline measurements, there were significant decreases in mean arterial blood pressure, mixed venous oxygen saturation, and right ventricular ejection fraction during maximal axial flow pump support and beta-blockade. No significant change in heart rate was observed at this time. Hemodynamic variables were normalized in the intensive care unit. All patients were separated from the Hemopump without inotropic support, and values of troponin-T, aspartate aminotransferase, and alanine aminotransferase were low postoperatively. All patients survived and were discharged from the hospital. CONCLUSION: The anesthetic protocol for patients undergoing surgery with a beating heart and the combined use of an axial flow pump and a short-acting beta-blocker is outlined. Multiple-vessel CABG on the beating heart was performed with maintenance of an acceptable hemodynamic situation. PMID- 10468257 TI - Comparison of cardiac output assessed by pulse-contour analysis and thermodilution in patients undergoing minimally invasive direct coronary artery bypass grafting. AB - OBJECTIVE: To investigate the precision and accuracy of continuous pulse contour cardiac output (PCCO) compared with intermittent transcardiopulmonary (TCPCO) and pulmonary artery thermodilution (TDCO) measurements in patients undergoing minimally invasive coronary bypass surgery (MIDCAB). DESIGN: Prospective, controlled, clinical study. SETTING: University hospital. PARTICIPANTS: Twelve patients undergoing MIDCAB. INTERVENTIONS: Thirty-six measurements of PCCO and thermodilution cardiac output (CO) were simultaneously performed after the start of surgery, during bypass grafting, and at the end of surgery. TCPCO and TDCO were simultaneously assessed by three injections of ice-cold saline randomly spread over the respiratory cycle. The pulse contour device was initially calibrated with an additional set of aortic thermodilution measurements. MEASUREMENTS AND MAIN RESULTS: Absolute values of CO ranged between 1.6 and 9.2 L/min. A close agreement among the three techniques was observed at all measurements. Mean bias between PCCO and TDCO and TCPCO was 0.003 L/min (2 SD of differences between methods = 1.26 L/min) and 0.27 L/min (2 SD of differences between methods = 1.16 L/min), respectively. The correlation coefficients were r2 = 0.90 for TCPCO versus PCCO and r2 = 0.88 for TDCO versus PCCO. CONCLUSION: The results of the present study show that compared with thermodilution CO, pulse contour analysis enables accurate measurement of continuous CO in patients undergoing MIDCAB. PMID- 10468258 TI - Support of mean arterial pressure during tepid cardiopulmonary bypass: effects of phenylephrine and pump flow on systemic oxygen supply and demand. AB - OBJECTIVE: To examine the effects of phenylephrine infusion and increases in pump flow on systemic oxygen supply and demand when they are used to support mean arterial pressure (MAP) during cardiopulmonary bypass (CPB). DESIGN: Prospective, unblinded study. SETTING: The animal cardiopulmonary laboratory at the Mayo Foundation (Rochester, MN). PARTICIPANTS: Twelve pigs. INTERVENTIONS: Twelve pigs had systemic oxygen delivery (DO2) and consumption (VO2) measured before CPB and then underwent CPB at 35 degrees C. During CPB, measurements of DO2 and VO2 were obtained at an MAP of approximately 50 mmHg and a pump flow of 2.2 L/min/m2. Thereafter, MAP was elevated to 70 mmHg either by increases in pump flow or by a phenylephrine infusion, and the balance between systemic oxygen supply and demand was reassessed. MEASUREMENTS AND MAIN RESULTS: Before CPB, DO2 was 375 +/- 83 mL/min/m2 and decreased with the onset of CPB mainly because of the effects of hemodilution. During CPB, with a pump flow of 2.2 L/min/m2 and an MAP of 53 mmHg, DO2 was 218 +/- 40 mL/min/m2. Increasing perfusion pressure to an MAP of 72 mmHg with phenylephrine and maintaining pump flow constant (2.2 L/min/m2) did not change DO2 (222 +/- 37 mL/min/m2), and the oxygen extraction ratio (OER) was increased relative to pre-CPB levels. In contrast, increasing MAP to 71 mmHg by increasing pump flow to 3.2 L/min/m2 resulted in a significantly greater DO2, and the OER normalized to the pre-CPB value. CONCLUSIONS: During CPB with conventional flow rates, DO2 is decreased. Supporting MAP with increases in pump flow better maintains DO2 than the administration of an alpha-agonist. PMID- 10468259 TI - Angiotensin II concentrations and gut mucosal perfusion in infants undergoing cardiopulmonary bypass. AB - OBJECTIVES: To determine whether hypothermic cardiopulmonary bypass (CPB) per se causes an increase in angiotensin II (A-II) concentration in infants and to investigate the relationship between A-II concentration and gut mucosal perfusion. DESIGN: Prospective, open, nonrandomized, observational study. SETTING: Children's teaching hospital. PARTICIPANTS: Thirty acyanotic infants requiring CPB. INTERVENTIONS: A-II concentrations were measured on six occasions before, during, and after CPB. An orogastric tonometer allowed intermittent calculations of gastric intramucosal pH (pHi). Gastric mucosal blood flow (flux) was monitored using a laser Doppler flowmeter. Ten infants acted as controls (group 1); 10 infants received captopril, 0.9 mg/kg orally, 45 minutes before induction of anesthesia (group 2), and 10 infants received enalaprilat, 0.06 mg/kg intravenously, just before CPB (group 3). MEASUREMENTS AND MAIN RESULTS: A II concentrations were abnormally high in 28 of 30 patients before CPB (median, 450 pg/mL (range, 83 to 5,787 pg/mL). A-II concentrations in groups 1 and 2 decreased during CPB, but values remained at twice normal levels throughout surgery (median, 171 to 198 pg/mL post-CPB). A-II concentrations remained normal (range, 52 to 120 pg/mL) during and after CPB in patients receiving enalaprilat (group 3). The authors found no significant correlation between A-II concentration and pHi or flux before, during, or after surgery. CONCLUSIONS: Acyanotic infants requiring cardiac surgery may have high perioperative concentrations of A-II. Hypothermic CPB is associated with a decrease in A-II concentration. Reductions in gut mucosal perfusion seen in some infants during hypothermic CPB are not related to increases in A-II concentrations. PMID- 10468260 TI - Differences among heart rate variability measures after anesthesia and cardiac surgery. AB - OBJECTIVE: To determine whether there are differences among measures of heart rate variability (HRV; traditional and nonlinear) after anesthesia and cardiac surgery. DESIGN: Prospective. SETTING: University hospital. PARTICIPANTS: Patients scheduled for cardiac surgery. INTERVENTIONS: None. Medical management was not varied as part of this study. MEASUREMENTS AND MAIN RESULTS: HRV was measured in 13 patients from electrocardiograms (ECGs) recorded before anesthesia, during anesthesia but before cardiac surgery, and on the first postoperative day. Anesthesia was induced with moderate-dose fentanyl. For each ECG, HRV was measured from series of 400 heartbeat intervals using standard deviation (SD), approximate entropy (ApEn), and point correlation dimension (PD2). Multivariate repeated-measures analyses of variance on ranks and Spearman correlations were performed. All HRV measures decreased significantly with anesthesia. Postoperatively, ApEn recovered to original values. PD2 and SD did not recover with consciousness and were significantly less than original values. Correlations among ApEn, PD2, and SD were weak. CONCLUSIONS: Nonlinear measures of HRV differ among themselves after anesthesia and cardiac surgery. The use of multiple nonlinear and traditional measures may improve the effectiveness of using HRV to assess the cardiovascular system. PMID- 10468261 TI - Monitoring of beta-receptor sensitivity in cardiac surgery. AB - OBJECTIVE: To determine the repeatability of the hemodynamic response to repeated isoproterenol challenge doses to validate the standardized isoproterenol sensitivity test as an index of cardiovascular beta-receptor function. DESIGN: Prospective, single-blind, nonrandomized clinical trial. SETTING: University department of cardiothoracic anesthesia. PARTICIPANTS: Twenty middle-aged men scheduled for primary elective coronary artery bypass surgery, 10 of whom had been treated with cardioselective beta1-antagonists for more than 3 months. INTERVENTIONS: After induction of anesthesia and baseline hemodynamic evaluation, cardiac beta-receptor sensitivity was estimated from the chronotropic/inotropic responses to four intravenous 4-microg isoproterenol bolus doses. MEASUREMENTS AND MAIN RESULTS: Baseline cardiovascular function and pharmacodynamic response to the four isoproterenol challenge doses were monitored with catheters in the radial and pulmonary arteries (thermodilution). Heart rate was continuously recorded and calculated from the electrocardiogram. Baseline hemodynamic status and response to the first 4 microg of isoproterenol were similar in the 10 patients treated with beta1-antagonists and the rest of the patients. In all 20 patients, heart rate response to the three subsequent isoproterenol challenge doses decreased progressively by 28%. CONCLUSION: The standardized isoproterenol sensitivity test is unreliable for clinical monitoring of cardiac beta adrenoceptor function. PMID- 10468262 TI - Port-access minimally invasive cardiac surgery in a patient without arms. PMID- 10468263 TI - Intraoperative rupture of the endoaortic clamp balloon in a patient undergoing port-access mitral valve repair. PMID- 10468264 TI - Management of an unusual presentation of aneurysm of the ductus arteriosus. PMID- 10468265 TI - Isolated left ventricular diverticulum in an adult. PMID- 10468266 TI - Pericardial mesothelioma: an incidental intraoperative diagnosis. PMID- 10468267 TI - Implantable cardioverter-defibrillators. PMID- 10468268 TI - Case 4--1999. Myocardial function improvement after hemofiltration in a septic patient. PMID- 10468269 TI - Pro: beta-blockers should be used in all patients undergoing vascular surgery. PMID- 10468270 TI - Con: beta-blockers should not be used in all patients undergoing vascular surgery. PMID- 10468271 TI - An abnormal transesophageal echocardiography finding after retrograde cardioplegia. PMID- 10468272 TI - A peril of minimally invasive surgery. PMID- 10468273 TI - Heparin-coated circuits: a new perspective--the boundary layer. PMID- 10468274 TI - Nicardipine infusion for radial artery spasm--a cautionary note. PMID- 10468275 TI - Nicardipine protocol for coronary artery bypass grafting using the radial artery: clinical and angiographic data. PMID- 10468276 TI - Paraplegia after coronary artery bypass graft surgery. PMID- 10468278 TI - Port-access minimally invasive cardiac surgery. PMID- 10468279 TI - Cardiac output measurements. PMID- 10468277 TI - Microbubble intravenous contrast during transesophageal echocardiography: agitated 0.9% saline versus agitated 0.9% saline with benzyl alcohol. PMID- 10468280 TI - Importance of monitoring both pressure waveforms during pulmonary artery catheterization. PMID- 10468281 TI - Adjuvant interferon in the treatment of melanoma. PMID- 10468282 TI - The leucocyte nadir, a predictor of chemotherapy efficacy? PMID- 10468283 TI - Prepubertal exposure to zearalenone or genistein reduces mammary tumorigenesis. AB - Prepubertal exposure to a pharmacological dose (500 mg kg(-1)) of the phyto oestrogen genistein can reduce the incidence and multiplicity of carcinogen induced mammary tumours in rats. However, such an exposure also disrupts the function of the hypothalamic-pituitary-gonadal axis, making it unsuitable for breast cancer prevention. We studied whether prepubertal exposure to genistein at a total body dose broadly comparable to the level typical of Oriental countries, approximately 1 mg kg(-1) body weight, affects mammary tumorigenesis. We also studied whether prepubertal exposure to zearalenone, a major source for phyto oestrogens in the USA, influences breast cancer risk. Prepubertal rats were treated between postnatal days 7 and 20, with 20 microg (approximately 1 mg kg( 1) body weight) of either genistein or zearalenone. Zearalenone exposure significantly reduced both the incidence and multiplicity of mammary tumours induced by 7,12-dimethylbenz(a)anthracene (DMBA). Genistein exposure significantly reduced tumour multiplicity, but not tumour incidence, when compared with vehicle-treated animals. Furthermore, 60% of the tumours in the genistein group were not malignant, while all the tumours analysed for histopathology in the vehicle and zearalenone groups were adenocarcinomas. A higher number of differentiated alveolar buds, and lower number of terminal ducts, were present in the DMBA-treated mammary glands of the phyto-oestrogen exposed rats. The concentration of oestrogen receptor (ER) binding sites after the DMBA treatment was low in the mammary glands of all groups but a significantly higher proportion of the glands in the zearalenone exposed rats were ER-positive (i.e. ER levels > or = 5 fmol mg(-1) protein) than the glands of the vehicle controls. Our data suggest that a prepubertal exposure to a low dose of either zearalenone or genistein may protect the mammary gland from carcinogen induced malignant transformation, possibly by increasing differentiation of the mammary epithelial tree. PMID- 10468284 TI - Fractionated gamma-irradiation renders tumour cells more responsive to apoptotic signals through CD95. AB - Signals through the CD95 surface receptor can specifically induce apoptosis. Some tumour cell lines are sensitive to CD95 signals, and insensitive cells can be converted to a sensitive phenotype if given appropriate treatment. To determine whether the apoptotic response of tumour cells to signalling through CD95 might be enhanced by ionizing irradiation, carcinoma cells were treated with either single-dose or fractionated gamma-irradiation. The response to treatment with an agonist anti-CD95 antibody was enhanced by pretreatment with either a single large dose or daily fractionated radiation. Fractionated irradiation induced cumulative and prolonged up-regulation of CD95 expression in cell lines bearing functional p53. Since two of four cell lines exhibiting heightened responsiveness to CD95-mediated signals following fractionated irradiation express mutant p53 and displayed little or no up-regulation of CD95, enhanced responsiveness did not correlate with p53 status and CD95 up-regulation. Continuous inhibition of CD95/CD95-ligand interactions during fractionated irradiation provided no protective effect to cells, arguing that autologous CD95/CD95-ligand interactions did not contribute to the direct lethal effect of irradiation. We conclude that fractionated gamma-irradiation provides an extended period of time when carcinoma cells are more responsive to CD95-mediated signals in vitro. PMID- 10468285 TI - Hypoxia-induced metastasis of human melanoma cells: involvement of vascular endothelial growth factor-mediated angiogenesis. AB - Tumour cells exposed to hypoxia have been shown to up-regulate the expression of vascular endothelial growth factor (VEGF). The purpose of the present work was to investigate whether hypoxia-induced VEGF up-regulation can result in increased metastatic efficiency of human melanoma cells. Two melanoma lines, one showing high (A-07) and the other showing low (D-12) VEGF secretion under aerobic conditions, were included in the study. Cell cultures were exposed to hypoxia (oxygen concentrations < 10 ppm) in vitro and metastatic efficiency, i.e. lung colonization efficiency, as well as transplantability and angiogenic potential were assessed in BALB/c-nu/nu mice Both cell lines showed significantly increased VEGF secretion under hypoxic conditions as measured by enzyme-linked immunosorbent assay The D-12 cells showed increased metastatic efficiency, transplantability and angiogenic potential following exposure to hypoxia. The metastatic efficiency increased with the duration of the hypoxia treatment and decreased with the time after reoxygenation. The A-07 cells on the other hand showed unchanged metastatic efficiency, transplantability and angiogenic potential following exposure to hypoxia. Both cell lines showed significantly decreased metastatic efficiency and angiogenic potential in mice treated with neutralizing antibody against VEGF. These results suggest that (a) VEGF is a limiting factor for the rate of angiogenesis in low but not in high VEGF expressing melanomas under normoxic conditions and (b) transient hypoxia might promote the development of metastases in low VEGF-expressing melanomas by upregulating the expression of VEGF and hence enhancing the angiogenic potential of the tumour cells. PMID- 10468286 TI - Autocrine and paracrine motility factors and their involvement in invasiveness in a human oral carcinoma cell line. AB - Invasive potentials of malignant cancer cells are regulated by cell motility factors. To examine the regulation of motility and invasiveness in oral squamous carcinoma, we investigated autocrine- and/or paracrine-acting cell motility factors, using a newly established human cell line (IF cells) from oral squamous cell carcinoma, which has highly invasive and metastatic characteristics. Conditioned medium derived from IF cells stimulated cell scattering and migration of GB-d1 gallbladder carcinoma cells, indicating that IF cells secreted cell motility factors. Using antibodies, IF-derived cell motility factors proved to be transforming growth factor (TGF)-alpha and TGF-beta1. Antibodies against TGF alpha and TGF-beta1 inhibited autonomous migration of the IF cells. On the other hand, in vitro invasion of IF cells was strongly enhanced by hepatocyte growth factor (HGF) but only slightly by TGF-alpha and TGF-beta1. The conditioned medium from fibroblasts enhanced in vitro invasion of IF cells, an event abrogated by anti-HGF antibody, but not by antibodies against TGF-alpha and TGF-beta1. Importantly, IF cells secreted a factor inducing HGF production in fibroblasts and the factor was identified as interleukin-1, which means that a mutual interaction exists between tumour cells and fibroblasts, as mediated by the HGF/HGF-inducer loop. These results indicate that IF cells utilize TGF-alpha and TGF-beta1 as autocrine-acting motility factors and HGF as a paracrine-acting motility factor, and that invasiveness of IF cells is particularly stimulated by HGF derived from stromal fibroblasts. Utilization of multiple cell motility/invasion factors that act in distinct pathways may confer highly invasive and metastatic potentials in IF oral squamous carcinoma cells. PMID- 10468287 TI - Antiproliferative effect of immunoliposomes containing 5-fluorodeoxyuridine dipalmitate on colon cancer cells. AB - We have investigated the antiproliferative action towards CC531 colon adenocarcinoma cells of target cell-specific immunoliposomes containing the amphiphilic dipalmitoyl derivative of 5-fluorodeoxyuridine (FUdR-dP). FUdR-dP incorporated in immunoliposomes caused a 13-fold stronger inhibition of CC531 cell growth in vitro, during a 72-h treatment, than FUdR-dP in liposomes without antibody, demonstrating that the prodrug is efficiently hydrolysed to yield the active drug, FUdR, intracellularly. The intracellular release of active FUdR was confirmed by determining the fate of 3H-labelled immunoliposomal FUdR-dP. Treatments shorter than 72 h with FUdR-dP in immunoliposomes resulted in anti tumour activities comparable to, or even higher than, that of free FUdR. The shorter treatments reflect more closely the in vivo situation and illustrate the potential advantage of the use of immunoliposomes over non-targeted liposomal FUdR-dP or free FUdR. Association of tumour cell-specific immunoliposomes with CC531 cells was up to tenfold higher than that of liposomes without antibody or with irrelevant IgG coupled, demonstrating a specific interaction between liposomes and target cells which causes an efficient intracellular delivery of the drug. Since biochemical evidence indicates a lack of internalization or degradation of the liposomes as such, we postulate that entry of the drug most likely involves the direct transfer of the prodrug from the immunoliposome to the cell membrane during its antigen-specific interaction with the cells, followed by hydrolysis of FUdR-dP leading to relatively high intracellular FUdR-levels. In conclusion, we describe a targeted liposomal formulation for the anticancer drug FUdR, which is able to deliver the active drug to colon carcinoma cells with high efficiency, without the need for the cells to internalize the liposomes as such. PMID- 10468288 TI - Increased cytotoxicity and bystander effect of 5-fluorouracil and 5-deoxy-5 fluorouridine in human colorectal cancer cells transfected with thymidine phosphorylase. AB - 5-Fluorouracil (5-FU) and 5'-deoxy-5-fluorouridine (5'-DFUR), a prodrug of 5-FU, are anticancer agents activated by thymidine phosphorylase (TP). Transfecting the human TP cDNA into cancer cells in order to sensitize them to these pyrimidine antimetabolites may be an important approach in human cancer gene therapy research. In this study, an expression vector containing the human TP cDNA (pcTP5) was transfected into LS174T human colon carcinoma cells. Eight stable transfectants were randomly selected and analysed. The cytotoxic effects of 5-FU and 5'-DFUR were higher in TP-transfected cells as compared to wild-type cells. The maximal decreases in the IC50 were 80-fold for 5-FU and 40-fold for 5'-DFUR. The increase in sensitivity to these pyrimidines of TP-transfected cells significantly correlated with the increase in both TP activity and TP expression. Transfected clone LS174T-c2 but not wild-type cells exhibited formation of [3H]FdUMP from [3H]5-FU. In addition the LS174T-c2 clone enhanced the cytotoxic effect of 5'-DFUR, but also that of 5-FU, towards co-cultured parental cells. For both anti-cancer agents, this bystander effect did not require cell-cell contact. These results show that both 5-FU or 5'-DFUR could be used together with a TP suicide vector in cancer gene therapy. PMID- 10468289 TI - Role of a proteolysis-inducing factor (PIF) in cachexia induced by a human melanoma (G361). AB - Human melanoma, G361, which induces cachexia in nude mice, has been shown to produce a proteolysis-inducing factor (PIF) of Mr 24000, which is immunologically identical to that isolated from a cachexia-inducing murine tumour (MAC16). Biosynthetic labelling of G361 cells using a combination of [35S]sulphate and [6 3H]glucosamine gave a single component of Mr 24000 after affinity chromatography employing a murine monoclonal antibody. The material contained both radiolabels and, after digestion with peptide N-glycosidase F, two fragments were produced of Mr 14000 and 10000 also containing both radiolabels. Digestion with O-glycosidase produced three fragments of Mr 14000, 6000 and 4000, the first two of which contained both radiolabels, while the third only contained 3H. This digestion pattern is the same as that previously observed with PIF from the MAC16 tumour and is commensurate with one N-linked sulphated oligosaccharide chain of Mr 10000, one O-linked sulphated oligosaccharide chain of Mr 6000 and a central polypeptide chain of Mr 4000 with some residual carbohydrate. When PIF from G361 cells was administered to female NMRI mice (20 g) a pronounced depression of body weight (1.36+/-0.36 g; P < 0.0001 from control) was observed over a 24 h period without a decrease in either food or water consumption. Body composition analysis showed a significant decrease in the non-fat carcass mass without a change in carcass fat or body water. This result suggests that depletion of lean body mass in mice bearing G361 melanoma arises from the production of PIF. PMID- 10468290 TI - Potentiation of the cytotoxicity of thymidylate synthase (TS) inhibitors by dipyridamole analogues with reduced alpha1-acid glycoprotein binding. AB - Dipyridamole has been shown to enhance the in vitro activity of antimetabolite anticancer drugs through the inhibition of nucleoside transport. However, the clinical potential of dipyridamole has not been realized because of the avid binding of the drug to the plasma protein alpha1-acid glycoprotein (AGP). Dipyridamole analogues that retain potent nucleoside transport inhibitory activity in the presence of AGP are described and their ability to enhance the growth inhibitory and cytotoxic effects of thymidylate synthase (TS) inhibitors has been evaluated. Three dipyridamole analogues (NU3026, NU3059 and NU3060) were shown to enhance the growth inhibitory activity of the TS inhibitor CB3717 and block thymidine rescue in L1210 cells. The extent of potentiation at a fixed analogue concentration (10 microM) was related to the potency of inhibition of thymidine uptake. A further analogue, NU3076, was identified, which was more potent than dipyridamole with a Ki value for inhibition of thymidine uptake of 0.1 microM compared to 0.28 microM for dipyridamole. In marked contrast to dipyridamole, inhibition of thymidine uptake by NU3076 was not significantly affected by the presence of AGP (5 mg ml(-1)). NU3076 and dipyridamole produced equivalent potentiation of the cytotoxicity of the non-classical antifolate TS inhibitor, nolatrexed, in L1210 cells with both compounds significantly reducing the LC90, by > threefold in the absence of salvageable thymidine. Thymidine rescue of L1210 cells from nolatrexed cytotoxicity was partially blocked by both 1 microM NU3076 and 1 microM dipyridamole. NU3076 also caused a significant potentiation of FU cytotoxicity in L1210 cells. These studies demonstrate that nucleoside transport inhibition can be maintained in the absence of AGP binding with the dipyridamole pharmacophore and that such analogues can enhance the cytotoxicity of TS inhibitors. PMID- 10468291 TI - Mathematical and experimental analysis of localization of anti-tumour antibody enzyme conjugates. AB - Considerable research has been aimed at improving the efficacy of chemotherapeutic agents for cancer therapy. A promising two-step approach that is designed to minimize systemic drug toxicity while maximizing activity in tumours employs monoclonal antibody (mAb)-enzyme conjugates for the activation of anticancer prodrugs. We present, analyse and numerically simulate a mathematical model based on the biology of the system to study the biodistribution, pharmacokinetics and localization properties of mAb-enzyme conjugates in tumour tissue. The model predictions were compared with experimental observations and an excellent correlation was found to exist. In addition, the critical parameters affecting conjugate half-life were determined to be the inter-capillary half distance and the antibody-antigen binding affinity. An approximation is presented relating the per cent injected dose per gram to inter-capillary half-distance and time. Finally, the model was used to examine various dosing strategies in an attempt to determine which regimen would provide the best biodistribution results. We compared the results of administering a uniform dose of fusion protein via bolus injection, multiple injections and continuous infusion. The model predicts that dosing strategy has little effect on the amount of conjugate that localizes in the tumour. PMID- 10468292 TI - Wheatgerm agglutinin-mediated toxicity in pancreatic cancer cells. AB - Lectin binding specificities for carbohydrate allow phenotypic and functional characterization of membrane-associated glycoproteins expressed on cancer cells. This analysis examined wheatgerm agglutinin binding to pancreatic cancer cells in vitro and the resulting toxicity. Membrane preparations of nine human pancreatic carcinoma cell lines were studied for lectin binding using wheatgerm agglutinin (WGA), concanavalin A (ConA) and phytohaemagglutinin-L (PHA-L) in a lectin blot analysis. Cell proliferation in vitro was measured by thymidine incorporation in the absence or presence of lectins at various concentrations. Sialic acid binding lectins or succinyl-WGA (succWGA) served as controls. WGA toxicity was tested after swainsonine or neuraminidase pretreatment. Binding and uptake of fluorescein-labelled lectins was studied under fluorescence microscopy. All pancreatic cell lines displayed high WGA membrane binding, primarily to sialic acid residues. Other lectins were bound with weak to moderate intensity only. Lectin toxicity corresponded to membrane binding intensity, and was profound in case of WGA (ID50 at 2.5-5 microg ml(-1)). WGA exposure induced chromatin condensation, nuclear fragmentation and DNA release consistent with apoptosis. Important steps for WGA toxicity included binding to sialic acid on swainsonine sensitive carbohydrate and lectin internalization. There was rapid cellular uptake and subsequent nuclear relocalization of WGA. In contradistinction to the other lectins studied, WGA proved highly toxic to human pancreatic carcinoma cells in vitro. WGA binding to sialic acid residues of N-linked carbohydrate, cellular uptake and subsequent affinity to N-acetyl glucosamine appear to be necessary steps. Further analysis of this mechanism of profound toxicity may provide insight relevant to the treatment of pancreatic cancer. PMID- 10468293 TI - Leucocyte nadir as a marker for chemotherapy efficacy in node-positive breast cancer treated with adjuvant CMF. AB - The purpose of this study was to examine the association between the leucocyte nadir and prognosis in breast cancer patients receiving adjuvant chemotherapy consisting of cyclophosphamide, methotrexate and fluorouracil (CMF). Three hundred and sixty-eight patients with node-positive breast cancer without distant metastases were treated with six cycles of adjuvant CMF. Some patients (n = 60) also received tamoxifen. All patients underwent surgery and received radiotherapy to the axillary and supraclavicular lymph nodes and the chest wall. The effect of leucopenia caused by CMF on distant disease-free survival (DDFS) and overall survival (OS) was assessed. A low leucocyte nadir during the chemotherapy was associated with a long DDFS in univariate analysis when tested as a continuous variable (the relative risk (RR) 1.3, 95% confidence interval (CI) 1.04-1.06, P = 0.02). Similarly, when the leucocyte nadir count was divided into tertiles, the patients who had the highest nadir values during the six-cycle treatment had worst outcome (RR 1.6, 95% CI 1.07-2.5, P = 0.02). However, in a multivariate analysis only the number of affected lymph nodes, tumour size, progesterone receptor status, surgical procedure, age and adjuvant tamoxifen therapy retained prognostic significance, whereas the leucocyte nadir count did not. A low leucocyte nadir during the adjuvant CMF chemotherapy is associated with favourable DDFS and it may be a useful biological marker for chemotherapy efficacy. PMID- 10468294 TI - Toxicity and feasibility of adjuvant high-dose interferon alpha-2b in patients with melanoma in clinical oncologic practice. AB - To assess the feasibility and toxicity profile of high-dose interferon alpha-2b (IFN-alpha-2b) in the adjuvant treatment of patients with cutaneous malignant melanoma outside the reference ECOG 1684 clinical trial, we conducted a prospective follow-up in an identical population of patients (cutaneous melanoma, T4 and/or N1) treated by intravenous IFN-alpha-2b:20 MIU m(-2), 5 days a week for 4 weeks; and subcutaneous:10 MIU m(-2), 3 times a week for 11 months. Thirty-six consecutive patients were considered in four different institutions. The frequency and severity of side-effects related to IFN-alpha, as well as the percentage of the planned dose given to patients, were identical to those reported in the initial report by ECOG. Fifty per cent and 47% of patients had a grade 3/4 WHO toxicity in the induction and consolidation phase respectively. A dose modification was necessary for 47.2% and 55.8% of the patients in the induction and consolidation phase respectively. The schedule and dose of high dose IFN-alpha-2b in the adjuvant treatment of cutaneous malignant melanoma, as reported by ECOG 1684, is feasible. The significant toxicity reported in ECOG 1684 was also seen in our patients. Nevertheless, this protocol will not be a 'standard' treatment until the publication of the ECOG 1690 trial. PMID- 10468295 TI - The effect of psychological interventions on anxiety and depression in cancer patients: results of two meta-analyses. AB - The findings of two meta-analyses of trials of psychological interventions in patients with cancer are presented: the first using anxiety and the second depression, as a main outcome measure. The majority of the trials were preventative, selecting subjects on the basis of a cancer diagnosis rather than on psychological criteria. For anxiety, 25 trials were identified and six were excluded because of missing data. The remaining 19 trials (including five unpublished) had a combined effect size of 0.42 standard deviations in favour of treatment against no-treatment controls (95% confidence interval (CI) 0.08-0.74, total sample size 1023). A most robust estimate is 0.36 which is based on a subset of trials which were randomized, scored well on a rating of study quality, had a sample size > 40 and in which the effect of trials with very large effects were cancelled out. For depression, 30 trials were identified, but ten were excluded because of missing data. The remaining 20 trials (including six unpublished) had a combined effect size of 0.36 standard deviations in favour of treatment against no-treatment controls (95% CI 0.06-0.66, sample size 1101). This estimate was robust for publication bias, but not study quality, and was inflated by three trials with very large effects. A more robust estimate of mean effect is the clinically weak to negligible value of 0.19. Group therapy is at least as effective as individual. Only four trials targeted interventions at those identified as at risk of, or suffering significant psychological distress, these were associated with clinically powerful effects (trend) relative to unscreened subjects. The findings suggest that preventative psychological interventions in cancer patients may have a moderate clinical effect upon anxiety but not depression. There are indications that interventions targeted at those at risk of or suffering significant psychological distress have strong clinical effects. Evidence on the effectiveness of such targeted interventions and of the feasibility and effects of group therapy in a European context is required. PMID- 10468296 TI - High-dose methotrexate in combination with interferons in the treatment of malignant pleural mesothelioma. AB - Twenty six patients with pleural mesothelioma of UICC stage I-IV excluding M1 disease (46% of whom had stage I disease and 38% stage III disease) were treated intravenously with high dose MTX (3 g) and calcium folinate rescue three times at intervals of 2 weeks and three times at intervals of 3 weeks. Natural interferon (IFN)-alpha (3 MIU days 2-10) and recombinant IFN-gamma1b (50 microg m(-2) on days 2, 6 and 10) were injected subcutaneously after each MTX dose. At the end of MTX treatment the IFNs were continued as maintenance therapy until disease progression. Seven partial responses were observed among 24 patients evaluable for response (response rate 29%, 95% confidence interval 13-51%). Median duration of response was 10 months (range 3-24 months). Median survival was 17 months and 1-year and 2-year survival rates 62% and 31% respectively. The toxicity of the chemo-immunotherapy was acceptable. Treatment was stopped in one patient who developed grade IV neurological toxicity. MTX dose reductions were rare (two patients with grade 1-2 renal toxicity). The combination of high dose MTX and IFN alpha and IFN-gamma is active against malignant pleural mesothelioma and well tolerated. The survival rates are encouraging. PMID- 10468297 TI - Phase I and pharmacokinetic study of DACA (XR5000): a novel inhibitor of topoisomerase I and II. CRC Phase I/II Committee. AB - DACA, also known as XR5000, is an acridine derivative active against both topoisomerase I and II. In this phase I study, DACA was given as a 3-h intravenous infusion on 3 successive days, repeated every 3 weeks. A total of 41 patients were treated at 11 dose levels between 9 mg m(-2) d(-1) and the maximum tolerated dose of 800 mg m(-2) day(-1). The commonest, and dose-limiting, toxicity was pain in the infusion arm. One patient given DACA through a central venous catheter experienced chest pain with transient electrocardiogram changes, but no evidence of myocardial infarction. At the highest dose levels, several patients also experienced flushing, pain and paraesthesia around the mouth, eyes and nose and a feeling of agitation. Other side-effects, such as nausea and vomiting, myelosuppression, stomatitis and alopecia, were uncommon. There was one minor response but no objective responses. DACA pharmacokinetics were linear and did not differ between days 1 and 3. The pattern of toxicity seen with DACA is unusual and appears related to the mode of delivery. It is possible that higher doses of DACA could be administered using a different schedule of administration. PMID- 10468298 TI - Concurrent conventionally factionated radiotherapy and weekly docetaxel in the treatment of stage IIIb non-small-cell lung carcinoma. AB - Docetaxel has shown remarkable radiosensitizing in vitro properties. In a previous phase I/II dose escalation study in non-small-cell lung cancer (NSCLC) we observed a high response rate after concomitant boost radiotherapy and weekly docetaxel. The maximum tolerated dose was 30 mg m(-2) week(-1). In the present phase II study we evaluated whether weekly docetaxel and conventionally fractionated radiotherapy could be better tolerated and equally effective in the treatment of locally advanced NSCLC. Thirty-five patients with T3, T4/N2, T3/M0 staged disease were recruited. Docetaxel (30 mg m(-2)) was given as a 30 min infusion once a week. Asthenia and radiation-induced oesophagitis were the main side-effects of the regimen enforcing 2-week treatment delay in 6/35 (17%) patients and minor delay (3-7 days) in another 11/35 (31%) patients. Neutrophil, platelet and haemoglobin toxicity was minimal, but pronounced lymphocytopenia was observed. Complete response (CR) of the chest disease was observed in 12/35 (34%) patients and partial response in 16/35 (46%). Although not statistically significant (P=0.19), a higher CR rate (8/18; 44%) was observed in patients who accomplished their therapy within the scheduled treatment time (44-47 days) as compared to patients that interrupted their treatment for several days due to treatment-related toxicity (CR 4/17; 23%). The overall survival and the local progression-free survival at 1 year was 48% and 60% respectively. We conclude that docetaxel combination with radiotherapy is a promising approach for the management of locally advanced NSCLC that results in high CR rate. Further trials with docetaxel-based radiochemotherapy should integrate accelerated radiotherapy together with cytoprotection. PMID- 10468299 TI - Phase II trial of gemcitabine, epirubicin and granulocyte colony-stimulating factor in patients with advanced pancreatic adenocarcinoma. AB - Although the novel cytidin analogue gemcitabine has shown superior anti-tumour activity than 5-fluorouracil in advanced pancreatic cancer, further improvements of therapeutic results are warranted. This goal might be achieved by combining gemcitabine with other active drugs. This trial evaluated the efficacy and tolerance of such a combination regimen with epirubicin and granulocyte colony stimulating factor (G-CSF) in patients with metastatic disease. Seventy patients with metastatic pancreatic adenocarcinoma were enrolled in this multicentre trial. Patients received 4-weekly courses of a combination regimen consisting of epirubicin 60 mg m(-2) given as intravenous bolus injection on day 1, gemcitabine 1000 mg m(-2) infused over 30 min on days 1, 8 and 15, and G-CSF administered at 5 microg kg(-1) day(-1) subcutaneously from days 2-6 during each cycle. The efficacy of treatment was assessed by conventional measures, i.e. objective response, progression-free and overall survival, as well as by analysis of clinical benefit response (defined as > or = 50% reduction in pain intensity, > or = 50% reduction in daily analgesic consumption, and/or > or = 20-point improvement in Karnofsky performance status that was sustained for > or = 4 consecutive weeks). Of 66 patients evaluable for objective response, one achieved complete and 13 partial remissions, for an overall response rate of 21% (95% confidence interval (CI), 12-33%); 27 additional patients (41%) had stable and 25 (38%) increasing disease. The median time to progression was 3.8 months. Median survival was 7.8 months, and the probability of surviving beyond 12 months was 21.2%. Out of 60 patients with tumour-related symptoms, who were considered evaluable for clinical benefit response, 26 (43%) experienced significant palliation. The median time to achieve a clinical benefit response was 7 weeks, and its median duration was 22 weeks. Chemotherapy was well-tolerated with leukopenia/granulocytopenia representing the most common and dose-limiting side effect. Gastrointestinal and other subjective toxicities were infrequent and generally rated minor. We conclude that the combination of gemcitabine, epirubicin and G-CSF seems to be an effective palliative treatment with only moderate toxic effects in patients with metastatic pancreatic adenocarcinoma. Our results in terms of objective and clinical benefit response, as well as survival seem to suggest an advantage over gemcitabine-monotherapy, though this remains to be confirmed in a randomized trial. PMID- 10468300 TI - Molecular detection of tumour DNA in serum and peritoneal fluid from ovarian cancer patients. AB - We have analysed DNA extracted from the serum and peritoneal fluid of 20 ovarian cancer patients for the presence of tumour-specific genetic alterations. The 20 patients included six with stage Ia disease. Using six polymorphic microsatellite loci we were able to detect novel alleles or loss of heterozygosity in 17/20 serum samples and 12/19 peritoneal fluid samples. Tumour-specific abnormalities were detected in the serum of all but one of the stage Ia cases. Half of the occurrences of loss of heterozygosity identified in primary tumour material were detectable in the serum samples. Novel alleles indicative of microsatellite instability were found in 3/6 patients with stage Ia disease but in only 1/14 of patients with more advanced disease. One of the eight patients in the control group displayed abnormalities in her serum DNA. The ease with which tumour specific alterations were detected in serum and peritoneal samples from ovarian cancer patients, using a panel of only six polymorphic microsatellite markers on four chromosomes, suggests that molecular detection methods could prove useful in the staging, monitoring and screening of this disease. PMID- 10468301 TI - Ki-67 is strongly prognostic in synovial sarcoma: analysis based on 86 patients from the Scandinavian Sarcoma group register. AB - In a study based on formalin-fixed paraffin-embedded material from 86 patients with primary synovial sarcoma located in the extremities or on the trunk wall, the prognostic importance of MIB-1 index, p53-expession and tumour size was analysed. Multivariate analysis identified two metastatic risk factors: increasing tumour size and MIB-1 > 9%. The 5-year metastasis-free survival-rate for patients with tumour size < or = 5 cm + MIB-1 < 10% was 0.83 (95% confidence interval (CI) 0.64-0.92) compared to 0.31 (95% CI 0.11-0.53) in cases with tumour size > 5 cm + MIB-1 > or = 10%. Our study shows that metastatic disease in synovial sarcoma is closely related to MIB-1 index. Using our model based on tumour size and MIB-1 index, cases with good and poor prognosis can easily be discriminated. Therefore our model can be used to identify patients who should be considered for adjuvant chemotherapy. PMID- 10468302 TI - Adrenal enlargement and failure of suppression of circulating cortisol by dexamethasone in patients with malignancy. AB - The aim of this study was to further elucidate the activity of the hypothalamo pituitary-adrenal (HPA) axis in patients with malignancy and to correlate this with the size of the adrenal glands. Fourteen patients with a variety of malignancies were studied prior to receiving cytotoxic chemotherapy. During routine staging computerized tomographic (CT) scans, the size of the body, medial and lateral limbs of the adrenal glands were measured and compared with those of a normal group of patients studied previously. Measurements of 09:00 h serum cortisol and plasma adrenocorticotropic hormone (ACTH) levels were made before and after the administration of dexamethasone (0.5 mg 6-hourly for 48 h) in addition to the peak cortisol response to i.v corticotropin releasing hormone (CRH). Overall, patients with malignancy had significantly larger adrenal glands than patients without malignancy; those with non-haematological malignancies had larger glands than patients with haematological malignancies. Following dexamethasone to suppress circulating cortisol levels, nine patients (64%) demonstrated abnormal resistance with cortisol levels > 50 nmol l(-1): mean value 294 nmol l(-1) (range 67-1147). Those patients who failed to suppress after dexamethasone had significantly larger adrenal glands than those that did suppress and tended to have non-haematological malignancies. ACTH levels were undetectable or low in three patients in whom it was measured and who did not suppress with dexamethasone. Following CRH, the cortisol levels were highest (823 and 853 nmol l(-1)) in two of these patients. Malignancy is associated with diffuse enlargement of the adrenal glands and resistance to dexamethasone-induced suppression of the HPA axis, which is not due to ectopic ACTH secretion. This disturbance of the normal control of the HPA axis is unexplained and its functional significance remains uncertain. PMID- 10468303 TI - Expression of membrane cofactor protein (MCP, CD46) in human liver diseases. AB - Membrane cofactor protein (MCP, CD46) is one of the complement regulatory proteins, and is widely distributed in human organs and protects cells from complement-mediated cytotoxicity. We analysed the distribution and the intensities of MCP in liver diseases and evaluated the role of MCP during hepatocarcinogenesis. Western blot analysis revealed that relative densities (density of the sample/density of the standard sample) of MCP in 27 HCC, 18 liver cirrhosis, nine chronic hepatitis and 12 normal liver were 0.63+/-0.23, 0.21+/ 0.07, 0.25+/-0.10 and 0.11+/-0.03 (mean+/-s.d.) respectively. MCP expression in hepatocellular carcinoma (HCC) was significantly higher than that in both liver cirrhosis and chronic hepatitis (P < 0.01). The difference in the tumour sizes, the grades of differentiation and viral marker status did not affect the expression. Immunohistological analysis revealed that MCP was distributed mainly in the basolateral membrane of the hepatic cord in non-cancerous liver, along with endothelial cells and bile duct cells. In HCC, the protein was observed on the membrane in a non-polarized fashion. These data suggest that HCC cells acquire the increased MCP expression in a development of HCC and may escape from tumour-specific complement-mediated cytotoxicity. PMID- 10468304 TI - A comparison of national cancer registry and direct follow-up in the ascertainment of ovarian cancer. AB - The National Health Service Central Register (NHSCR) and direct follow-up were used to document ovarian and fallopian tube cancers in 22000 women from 1986 to 1993. Direct follow-up identified 47/49 cases (96%) and the NHSCR 38/49 (78%). NHSCR ascertainment was incomplete and direct follow-up provided additional information. These findings have implications for interpretation of national cancer statistics and for use of the NHSCR in research trials. PMID- 10468305 TI - Risk of cervical cancer is not increased in Chinese carrying homozygous arginine at codon 72 of p53. AB - Homozygous arginine at codon 72 (HA72) of p53 was found in 22% of normal cervices and 30.0% of cervical cancers and no significant difference was detected between normal and cervical cancer with or without HPV 16/18. There was no correlation between HA72 and risk of cervical cancer in Chinese. PMID- 10468306 TI - Diabetes mellitus, other medical conditions and familial history of cancer as risk factors for pancreatic cancer. AB - In a population-based case-control study of pancreatic cancer conducted in three areas of the USA, 484 cases and 2099 controls were interviewed to evaluate the aetiologic role of several medical conditions/interventions, including diabetes mellitus, cholecystectomy, ulcer/gastrectomy and allergic states. We also evaluated risk associated with family history of cancer. Our findings support previous studies indicating that diabetes is a risk factor for pancreatic cancer, as well as a possible complication of the tumour. A significant positive trend in risk with increasing years prior to diagnosis of pancreatic cancer was apparent (P-value for test of trend = 0.016), with diabetics diagnosed at least 10 years prior to diagnosis having a significant 50% increased risk. Those treated with insulin had risks similar to those not treated with insulin (odds ratio (OR) = 1.6 and 1.5 respectively), and no trend in risk was associated with increasing duration of insulin treatment. Cholecystectomy also appeared to be a risk factor, as well as a consequence of the malignancy. Subjects with a cholecystectomy at least 20 years prior to the diagnosis of pancreatic cancer experienced a 70% increased risk, which was marginally significant. In contrast, subjects with a history of duodenal or gastric ulcer had little or no elevated risk (OR = 1.2; confidence interval = 0.9-1.6). Those treated by gastrectomy had the same risk as those not receiving surgery, providing little support for the hypothesis that gastrectomy is a risk factor for pancreatic cancer. A significant 40% reduced risk was associated with hay fever, a non-significant 50% decreased risk with allergies to animals, and a non-significant 40% reduced risk with allergies to dust/moulds. These associations, however, may be due to chance since no risk reductions were apparent for asthma or several other types of allergies. In addition, we observed significantly increased risks for subjects reporting a first-degree relative with cancers of the pancreas (OR = 3.2), colon (OR = 1.7) or ovary (OR = 5.3) and non-significantly increased risks for cancers of the endometrium (OR = 1.5) or breast (OR = 1.3). The pattern is consistent with the familial predisposition reported for pancreatic cancer and with the array of tumours associated with hereditary non-polyposis colon cancer. PMID- 10468307 TI - Cytochrome P4501A1 polymorphism as a susceptibility factor for breast cancer in postmenopausal Chinese women in Taiwan. AB - The incidence of breast cancer has been greatly increasing in Taiwan over the past two decades. Since cytochrome P4501A1 (CYP1A1) is involved in the metabolism of environmental carcinogens or oestrogen, we hypothesized that CYP1A1 genetic polymorphism may be a susceptibility factor for breast cancer. This hypothesis was evaluated in this case control study of 150 breast cancer patients and 150 healthy controls among Chinese women. Two CYP1A1 polymorphisms were studied, one containing a new Msp1 site and the other located in axon 7 and resulting in the replacement of an isoleucine (Ile) residue by a valine (Val). After simultaneously considering the known or significant risk factors for breast cancer, including the age of study participants, positive family history of breast cancer, early menarche (< or = 13 years), nulliparity and late first full term pregnancy (> or = 30 years), hormone replacement therapy and smoking, the CYP1A1 Msp1 polymorphism was found to be a significant factor in determining the risk of breast cancer. The homozygous variant was the most susceptible genotype with an adjusted odds ratio of 1.98 (95% confidence interval (CI) = 1.01-3.99) compared with the non-homozygous variants (the homozygous wild-type and the heterozygous variant). In contrast, the CYP1A1 Ile/Val polymorphism was not significantly associated with breast cancer development (adjusted OR = 1.07, 95% CI = 0.64-1.78). Interestingly, the Msp1 polymorphism was especially significant in postmenopausal women, but not in premenopausal women. Further stratification analysis in postmenopausal women who were non-smokers and with no history of hormone replacement therapy showed the cancer risk due to the Msp1 variant to be more significant in women with early menarche. We conclude that CYP1A1 polymorphism is a susceptibility factor for breast cancer in postmenopausal Chinese women in Taiwan. Further study with a large sample size should be considered to address issues of interactions between CYP1A1 and other risk factors. PMID- 10468308 TI - Pre- and perinatal risk factors for childhood leukaemia and other malignancies: a Scottish case control study. AB - A case control study of Scottish children aimed to identify risk factors for leukaemia and other cancers operating in the prenatal environment, during delivery and neonatally. Cases (0-14 years) were age-and sex- matched to two population-based controls and details abstracted from the mother's hospital obstetric notes. Analyses of 144 leukaemias (124 acute lymphoblastic leukaemias (ALL)), 45 lymphomas, 75 central nervous system (CNS) tumours and 126 'other solid tumours' were conducted using conditional logistic regression. The presence of a neonatal infection significantly reduced the risk of ALL (odds ratio (OR) 0.49, 95% confidence interval (CI) 0.26-0.95), particularly in 0- to 4-year-olds. Positive swab tests confirmed 47% of ALL cases with any infection and 46% of controls. This is consistent with the hypothesis that early exposure to infections may reduce the risk of childhood ALL. Asphyxia at birth significantly increased the risk of leukaemia, which was accounted for by ALL. For the 'other solid tumours' higher levels of maternal education were inversely associated with risk (OR 0.59, 95% CI 0.37-0.94) but positively associated with antibiotics (OR 2.16 95% CI 1.10-4.25) and respiratory tract infections (OR 14.1, 95% CI 1.76 113.7) in pregnancy. No obvious plausible patterns of risk were detected either within or across disease subgroups. PMID- 10468309 TI - Lifetime exercise activity and breast cancer risk among post-menopausal women. AB - Lifetime exercise activity has been linked to breast cancer risk among young women. However, no study has specifically evaluated whether lifetime exercise activity is related to the breast cancer risk of post-menopausal women. We conducted a population-based case-control study of post-menopausal white women (1123 newly diagnosed cases and 904 healthy controls) aged 55-64 who lived in Los Angeles County, California, USA to evaluate this relationship. Although neither exercise activity from menarche to age 40 years, nor exercise after age 40 separately predicted breast cancer risk, risk was lower among women who had exercised each week for at least 17.6 MET-hours (metabolic equivalent of energy expenditure multiplied by hours of activity) since menarche than among inactive women (odds ratio (OR) = 0.55; 95% confidence interval (CI) 0.37-0.83). Exercise activity was not protective for women who gained considerable (> 17%) weight during adulthood. However, among women with more stable weight, breast cancer risk was substantially reduced for those who consistently exercised at high levels throughout their lifetime (OR = 0.42; 95% CI 0.24-0.75), those who exercised more than 4 h per week for at least 12 years (OR = 0.59; 95% CI 0.40 0.88), and those who exercised vigorously (24.5 MET-hours per week) during the most recent 10 years (OR = 0.52; 95% CI 0.32-0.85). Strenuous exercise appears to reduce breast cancer risk among post-menopausal women who do not gain sizable amounts of weight during adulthood. PMID- 10468310 TI - Risk factors for testicular germ cell tumours by histological tumour type. United Kingdom Testicular Cancer Study Group. AB - There are two main histological groups of testicular germ cell tumours, which may have different risk factors. Some authors have analysed potential risk factors by histological group but few consistent differences have been identified. In this paper we examine risk factors for pure seminoma and other tumours using data from the United Kingdom case control study of testicular cancer. Seven hundred and ninety-four cases were included in the study, each with a matched control; 400 cases had pure seminoma tumours, and 394 had other testicular tumours. The risk of seminoma associated with undescended testis was slightly higher than that for other tumours (odds ratio of 5.3 compared with 3.0). When split at the median age at diagnosis, this difference was greater in men aged 32 and over (odds ratio of 11.9 compared with 5.1) than in the younger men (3.0 compared with 2.5). Risks associated with testicular or groin injuries were higher in the non-seminoma group, as was the risk for a history of sexually transmitted disease. The protective effect of a late puberty was more marked for tumours of other histologies. Some differences were also detected for participation in sports. Whilst some of the differences detected may have arisen by chance, the stronger association between undescended testis and pure seminoma has been identified by a number of other studies and may reflect a genuine difference in aetiology. PMID- 10468311 TI - Quantification and prognostic relevance of angiogenic parameters in invasive cervical cancer. PMID- 10468312 TI - Effects of tiapride on electroencephalograms and cognitive functions in the elderly. AB - Tiapride is a substituted benzamide with selective dopamine D2 and D3-antagonist properties which appears to have preferential affinity for extra-striatal dopamine receptors. Tiapride is used in the treatment of agitation, aggressiveness and anxiety in the elderly. To define the effects of a single dose of tiapride 100 mg on psychomotor performance and cognitive functions and electroencephalogram (EEG), a randomized, double-blind, three-way crossover, placebo-controlled study using lorazepam 1 mg as a positive control was carried out in 12 elderly individuals (six women and six men, mean age +/- SD: 69 +/- 3 years). A 1-week wash-out interval was allowed between each administration. Psychomotor and cognitive functions were assessed using both objective [EEG, critical flicker fusion, simple reaction time, tapping, body sway, continuous performance task (CPT), digit symbol substitution test, Sternberg memory scanning and a learning memory test using word lists] and subjective (visual analogue scales) measures before and up to 6 h after dosing. Tiapride was devoid of any detrimental or sedative effects on EEG and all of the performance tasks used and did not impair memory compared with-placebo. In contrast, a single dose of lorazepam produced significant deleterious effects on psychomotor performance (decrease in tapping and in sustained attention (CPT) and an increase in reaction time and body sway), and sedative effects on EEG (significant increase in delta and decrease in alpha waves) as well as significant impairment in working memory (Sternberg) and anterograde amnesia (decrease in immediate and delayed free recall) up to 6 h after dosing compared with placebo and tiapride. In conclusion, the present study showed that in contrast to lorazepam 1 mg there is no evidence to suggest that a single dose of tiapride 100 mg has any sedative and amnestic effects in the elderly which may interfere with everyday life activities. PMID- 10468313 TI - Safety of amisulpride (Solian): a review of 11 clinical studies. AB - We assessed the overall safety profile of amisulpride based on the results from 11 clinical studies performed in patients suffering from schizophrenia with predominance of positive or negative symptoms. A total of 1933 patients were randomly assigned to treatment with amisulpride (n = 1247) or haloperidol (n = 309), risperidone (n = 113), flupentixol (n = 62) and placebo (n = 202). Safety data collection was performed using open reporting, UKU scales or specific extrapyramidal side-effect scales; electrocardiogram recording and vital signs examination; laboratory data collection. Amisulpride demonstrated a satisfactory global safety profile in the range of doses usually prescribed. The number of patients having at least one extrapyramidal side-effect was higher in haloperidol patients compared with both amisulpride and risperidone patients (50% versus 30% in the two latter groups). For endocrine events, a similar rate was observed between amisulpride and risperidone groups (4% versus 6%, respectively) versus 1% in the haloperidol group. Electrocardiogram results were satisfactory, confirmed by the absence of cardiovascular events. The overall laboratory safety profile of amisulpride did not show clinically relevant abnormalities in liver function tests nor haematological abnormalities. Our extensive clinical data confirm the satisfactory safety profile of amisulpride which is superior to standard reference compounds. PMID- 10468314 TI - Reduced haloperidol does not interfere with the antipsychotic activity of haloperidol in the treatment of acute schizophrenia. AB - The aim of this study was to investigate the effect of reduced haloperidol, the main metabolite of the antipsychotic drug haloperidol, on psychopathology improvement and extrapyramidal adverse effects in acute schizophrenia. The steady state pharmacokinetics of reduced haloperidol was studied. Serum concentrations of reduced haloperidol (C(RH)) and haloperidol (C(H)) were measured in an open clinical trial over 6 weeks of treatment in 57 acutely schizophrenic patients. Psychopathology was measured by the Brief Psychiatric Rating Scale and several subscales. The assay of extrapyramidal adverse effects was conducted by means of the Extrapyramidal Symptom Rating Scale. A significant serum concentration therapeutic effect relationship (SCTER) of haloperidol of the same data has been demonstrated. In our study, the influence of the metabolite reduced haloperidol on the antipsychotic activity of haloperidol was analysed by means of regression analysis of the residuals of the SCTER of haloperidol with C(RH). In addition, the steady-state pharmacokinetics of reduced haloperidol and direct relationships between C(RH) and the metabolite ratio C(RH)/C(H) with psychopathology improvement and extrapyramidal adverse effects were investigated. Reduced haloperidol was not found to interfere with the antipsychotic action of the parent drug. Patients with elevated C(RH) or elevated metabolite ratio C(RH)/C(H) did not show consistently lower clinical improvements compared with the fitting curve of the SCTER of haloperidol and therefore no significant relationship between C(RH) and the residuals of the SCTER of haloperidol was detected. Furthermore, no significant result was found in the analysis of the direct relationships of C(RH) and C(RH)/C(H) with clinical variables which, for example, indicate decreased outcome with increased C(RH). In contrast, because of the pharmacokinetic dependence of C(RH) and C(H), a trend for a bisigmoidal relationship with C(RH) emerged for some outcome variables which was traced as an epiphenomenon from the bisigmoidal SCTER of the parent drug (e.g. change of hostility after 3 weeks). No relationship of reduced haloperidol with extrapyramidal adverse effects could be detected. It is concluded that serum concentrations of reduced haloperidol are of minor value for the interpretation of data of therapeutic drug monitoring of haloperidol in patients with acute schizophrenia. Reduced haloperidol does not act as a 'false neuroleptic'. PMID- 10468315 TI - The effect of long-term antipsychotic treatment on the body weight of patients suffering from chronic schizophrenia: clozapine versus classical antipsychotic agents. AB - The aim of this study was to evaluate the effect of long-term clozapine treatment on body weight changes in neuroleptic-resistant chronic schizophrenic patients and to compare it with that of classical antipsychotic agents. The body mass index (BMI) of 96 neuroleptic-resistant chronic schizophrenic patients was calculated before the beginning and after long-term (mean +/- SD 1.7 +/- 1.3 years) clozapine treatment. These data were compared to the BMI of 98 chronic schizophrenic patients maintained on classical antipsychotic agents for a similar duration (mean +/- SD 1.9 +/- 1.6 years). A significant elevation in BMI was detected in both groups during these periods (P < 0.0001 versus baseline, for both groups). The change in BMI (delta BMI) was similar in both groups (P < 0.9). We conclude that the increase in body weight caused by long-term (> 6 months) clozapine treatment is comparable to that obtained following long-term classical antipsychotic agents treatment. PMID- 10468316 TI - Adjunctive nefazodone in neuroleptic-treated schizophrenic patients with predominantly negative symptoms: an open prospective pilot study. AB - A combination of nefazodone with a conventional neuroleptic would lead to a serotonin (5-HT)2 and D2 receptor blockade resembling that of an atypical neuroleptic, with an additional increase of 5-HT (and noradrenaline) turnover. This may be of benefit in some cases of schizophrenia. In this study, eight patients with schizophrenia with predominantly negative and/or depressive symptoms underwent an open prospective 26-week trial with nefazodone, added to conventional neuroleptics. The total Positive and Negative Syndrome Scale (PANSS) and the Montgomery-Asberg Depression Rating Scale (MADRS) scores (the last observations carried forward, LOCF) significantly (P < 0.05) decreased in these eight patients by a mean of 31% and 63%, respectively, mainly within the first 6 weeks. Positive symptoms, observed in three patients and panic attacks in two patients disappeared entirely. The doses of neuroleptics, stable during the first 6 weeks of the trial, subsequently were able to be decreased by 28%. Extrapyramidal symptoms noticeably improved during the phase of stable neuroleptic dose regimen. Of the three patients who discontinued the trial prematurely (after 14 weeks or more), only one evidenced a nefazodone-related adverse event. Adjunctive nefazodone may be a useful treatment option in this patient population, but additional studies are recommended. PMID- 10468317 TI - Venlafaxine in social phobia: a study in selective serotonin reuptake inhibitor non-responders. AB - The study aimed to evaluate the clinical response to venlafaxine in social phobia in 12 patients who were non-responders to selective serotonin reuptake inhibitors, and to assess how the response could be influenced by the comorbidity in Axis II with avoidant personality disorder (APD). The duration of the study was of 15 weeks using open flexible doses regimen in individuals with or without concomitant APD. The venlafaxine dose ranged from 112.5 mg/day to 187.5 mg/day. Venlafaxine improves social phobia and/or APD symptomatology, as demonstrated by decreasing Liebowitz Social Anxiety Scale total scores (P < 0.05). In fact, venlafaxine significantly reduced the avoidant behaviour and specific sociophobic aspects, while notably improving the depression dimension and the basic anxiety symptoms. With regard to tolerability, the profile of venlafaxine was satisfactory with the main side-effects being nausea, headache and anxiety. PMID- 10468318 TI - Prevalence of headache syndromes in panic disorder. AB - We investigated the prevalence of headache in a group of patients attending a psychiatric clinic because suffering from panic disorder, according to DSM-IV criteria. The psychopathological assessment was performed with the 'Panic Disorder/Agoraphobia Questionnaire' and the presence of headache was evaluated according to the criteria of the International Headache Society. The results showed that two-thirds of patients met the criteria for a diagnosis of headache, with migraine without aura being the most frequent form, followed by tension headache, while two patients only were affected by migraine with aura. When we compared panic patients with and without headache, those with headache had a longer duration of panic disorder, a higher number of attacks and a heavier family loading for panic disorder and headache. This suggests that the comorbidity of headache with panic disorder renders this condition more severe and possibly responsive to different treatments compared to panic disorder alone. PMID- 10468319 TI - The use of mirtazapine in a group of 11 patients following poor compliance to selective serotonin reuptake inhibitor treatment due to sexual dysfunction. AB - Mirtazapine, a noradrenergic and specific serotonergic antidepressant, displays strong serotonin (5-HT)2 blocking properties, which may be related to lack of sexual dysfunction. In our open-label study, after a wash-out period of 4-14 days, mirtazapine (30-45 mg/day) was administered for 6 weeks to six male and five female patients who discontinued treatment with selective serotonin reuptake inhibitors (SSRIs) because of sexual dysfunction. The patients were moderately depressed, with baseline Hamilton Depression Rating Scale (17-item HAMD) scores between 19 and 24, and none of them experienced any sexual dysfunction prior to SSRI treatment. Efficacy was assessed weekly by 17-item HAMD, and adverse events were registered at the same time points. All patients completed the study. After 6 weeks of treatment, the individual 17-item HAMD scores were between 5 and 9, indicating significant improvement in depressive symptoms. None of the patients reported any sexual dysfunction symptoms. Other adverse events, mild and transient in nature, were reported only by three patients (somnolence in two, and weight gain in one patient). In conclusion, treatment with mirtazapine was effective in patients who are unable to tolerate SSRIs because of sexual dysfunction and demonstrated no effect on sexual function. PMID- 10468320 TI - Nefazodone-induced clitoral priapism. AB - A case is presented of painful priapism of the clitoris lasting 3 days, with a strong temporal association with the administration of nefazodone hydrochloride. Priapism has been described as a rare side-effect of drugs with high alpha1 adrenergic blocking potential. However, the alpha1-adrenergic blocking potential of nefazodone is moderate. Drug-induced clitoral priapism has been described for citalopram, bromocriptine, fluoxetine and trazodone combined with fluoxetine. To our knowledge, this is the first report in the literature about priapism of the clitoris associated with nefazodone. PMID- 10468321 TI - Noradrenaline and serotonin in depression: consensus and controversies. PMID- 10468322 TI - Physiology to functionality: the brain and neurotransmitter activity. AB - The monoamine hypothesis of depression proposes that the biological basis for depression is a deficiency in the neurotransmitters serotonin or noradrenaline, or both. Over the past 30 years this hypothesis has been refined as more experimental and clinical evidence has emerged. The selective serotonin reuptake inhibitors in particular have made a significant contribution to our understanding of the role of serotonin in depression. Our knowledge of the noradrenergic system is less complete, mainly because, until recently, no equivalent agents were available for this system. With the advent of reboxetine, the first selective noradrenaline reuptake inhibitor, attention is again focusing on the role of noradrenaline in depression. To an extent, the action of the selective inhibitors can be predicted through knowledge of the neuroanatomy of the central and peripheral nervous systems. With regard to depression, the most important pathways are those of the serotonergic and noradrenergic neurones projecting to the prefrontal cortex, from the raphe nucleus and locus coeruleus, respectively. However, increasing the levels of the monoamines in the central nervous system affects many other pathways and a wide range of serotonin and adrenergic receptors, leading to a multiplicity of potential beneficial and adverse events. In addition, the complex intracellular responses are beginning to be examined, again with the aid of the selective antidepressants. PMID- 10468323 TI - Antidepressants and the brain. AB - The pathophysiology and effects of antidepressants in the brain are still poorly understood. While it is generally accepted that increasing the levels of monoamine in the brain is an effective way to alleviate depression, the precise neurobiological mechanisms are unclear. The evidence that monoamine function is impaired in individuals with depression is largely indirect. However, the neurotransmitter depletion model allows a more direct investigation of the role of the monoamines. In this model, tryptophan depletion is used to lower levels of serotonin and alpha-methylparatyrosine is used to induce catecholamine depletion in the brain. Studies have shown that such depletion transiently reverses antidepressant responses in the majority of patients, the response being dependent on the type of antidepressant used. However, depletion in unmedicated patients with depression did not worsen the depressive symptoms, neither did it cause depression in healthy subjects with no history of mental illness. The cause(s) of depression therefore appears to be more complex than simply a reduction in levels of monoamine or diminished function in these systems. The pathophysiology of depression may relate to dysfunction in brain areas modulated by monoamine systems. Antidepressant drugs may mediate their effects by causing adaptive changes in neurones localised in these brain areas. PMID- 10468324 TI - Predicting response: serotonin reuptake inhibition. AB - The introduction of selective serotonin reuptake inhibitors (SSRIs) has helped clarify our understanding of the actions of serotonin in depression. The serotonergic network of the central and peripheral nervous systems comprises many different pathways and at least 14 distinct serotonin receptors. By using our knowledge of neuroanatomy and the pharmacology of SSRIs, it is possible to predict many of the therapeutic and non-therapeutic effects of modulating serotonin levels. The efficacy of SSRIs in mild to moderate depression is well established but their efficacy in severe depression varies, compared with tricyclic antidepressants. The SSRIs also have predictable effects in a range of other affective disorders including obsessive-compulsive disorder, anxiety and bulimia, the efficacy varying between the different SSRIs. The adverse event profile is different from that of the tricyclic antidepressants and again is predictable through an understanding of the different serotonergic pathways. PMID- 10468325 TI - Predicting response: noradrenaline reuptake inhibition. AB - For the past decade, the role of noradrenaline in depression has been somewhat neglected in favour of serotonin. This is largely because of the advent of the selective serotonin reuptake inhibitors, which have facilitated clinical and experimental observation of the roles of serotonin. Until now, no such tools have been available to study the noradrenergic system. However, the recent development of reboxetine, the first selective noradrenaline reuptake inhibitor, has allowed clinical investigation of the role of the noradrenergic system in different aspects of depressive disorders. In clinical trials, the use of reboxetine has shown that selective noradrenaline reuptake inhibition is an effective approach to alleviating depression. It is more effective than placebo and at least as effective as desipramine, imipramine and fluoxetine in the short term. In addition, its efficacy is maintained in patients with severe depression and in those receiving long-term maintenance treatment. Reboxetine is very well tolerated, as predicted from its pharmacological profile, having fewer anticholinergic side-effects than imipramine or desipramine. Compared with fluoxetine, patients treated with reboxetine experienced less nausea and sexual dysfunction, adverse events that are common among those taking selective serotonin reuptake inhibitors. Adverse events predicted by the neuroanatomy of the noradrenergic system, such as tremor and cardiovascular effects, occurred less frequently than expected. Clinical experience with reboxetine challenges our current knowledge of the role of noradrenaline in depression and questions existing evidence based on studies with noradrenergic tricyclic antidepressants. Selective noradrenaline reuptake inhibition, as exemplified by reboxetine, therefore offers a significant improvement in antidepressant pharmacotherapy, and an opportunity to increase our understanding of the role of noradrenaline in depression. PMID- 10468326 TI - From symptoms to social functioning: differential effects of antidepressant therapy. AB - Significant impairments in social functioning frequently occur simultaneously with depressive symptoms. The implications of such impairments extend beyond the depressed individual to their family, friends and society at large. Classical rating scales such as the Hamilton rating scale for depression primarily assess the core symptoms of depression. A range of rating scales are available, both self-reporting and administered by clinician; however, many have been criticised for their unspecified conceptual background and for being complex and time consuming. While antidepressants in general appear to improve social functioning, no clear advantage for any single class of agent has been reported. Recently, a new self-report rating scale, the Social Adaptation Self-evaluation Scale, has been developed and used to compare the novel selective noradrenaline reuptake inhibitor, reboxetine, with the selective serotonin re-uptake inhibitor, fluoxetine. The noradrenergic agent, reboxetine, was shown to be significantly more effective in improving social functioning than the serotonergic agent, fluoxetine. These findings are consistent with previous observations that noradrenaline may preferentially improve vigilance, motivation and self perception. PMID- 10468327 TI - The use of public health approaches in mental health programmes. AB - This paper underlines the usefulness of public health approaches to programme planning and evaluation in the field of psychiatry. The use of these approaches is illustrated by an assessment of the public health importance of depressive disorders and by a review of public health criteria for the evaluation of treatment interventions in psychiatry. The author stresses that the use of public health approaches in psychiatry is necessary because it will help in national planning and in increasing the priority which the authorities and the general public give to mental health programmes. PMID- 10468328 TI - Sharing the responsibilities: the roles of the government. AB - Developments in psychiatry have been steered by a number of factors influencing decisions taken by the politicians. Some of the more typical factors that could be mentioned are: political attitudes; trend phenomena in the population; economic aspects; and evidence-based knowledge. Bearing in mind the results of developments in psychiatry during the last decades, psychiatrists are encouraged to live up to their responsibility by focusing on scientific results, when giving their advice to politicians. Hopefully political initiatives will, in the future, primarily be based on evidence about psychiatric treatment. PMID- 10468329 TI - Results of treatment of schizophrenia: is the glass half full or half empty? AB - On the basis of data from naturalistic studies it is shown that the actual outcome in schizophrenia is by far not as good as it could be. Thus, for example, the relapse rates of schizophrenic patients lie 200% above the value that could actually be achieved according to today's state of knowledge. This results in extra costs of several billion dollars each year. The causes of this great discrepancy between possible and actual outcome are hardly known, since psychiatric researchers have previously not been very interested in the outcome of normal care patients. But patients, carers and fund holders expect that psychiatrists should rapidly close the gap between the actual and the possible outcome. They will succeed only if psychiatric researchers become more interested in the normal care situation, perform more naturalistic studies, more compliance research, and more studies into medical decision making. PMID- 10468330 TI - Theory and practice in the management of depressive disorders. AB - Scientific, professional, and governmental bodies have in recent years intensified efforts to formulate guidelines for the treatment of depression with the goal that medical practice should comply with these standards and thus improve patient care. 'Evidence-based medicine' demands that medical practice should explicitly refer to the best available scientific knowledge. Empirical data on the epidemiology of care suggest that there are major differences between guidelines and routine practice. Failure to detect mental disorders range from 45 to 90%. Only 1 in 10 of those suffering from depression receive adequate treatment. Treatment is insufficient as prescribed dosages are routinely too low, as up to 50% of treatments are ended prematurely and because patient non compliance interferes with proper treatment application. Psychological treatment is provided only in 60% of cases in need and is mostly unspecific, i.e. listening or giving advice. Furthermore, there are large regional variations in prescribing patterns which show that treatment is influenced by many non-medical factors. Empirical studies show that giving information to physicians or even measures to improve recognition alone are not sufficient to make changes in treatment behaviour. Only if measures are taken to improve treatment interventions directly then, at least for more severe cases, could better illness outcome be shown. If one wants to understand therapist non-compliance with treatment guidelines then one first has to understand the process of medical decision making. Action theory provides a model which brings together theoretical and experimental knowledge, goal setting, medical and emotional assessment, and operational procedures. On the basis of this theory, future guidelines should specify the setting for which they claim to be valid, they should be empirically tested for their feasibility and effects on patient outcome, and they should give proper attention to non medical factors which have major impact on medical practice. PMID- 10468331 TI - Improvement of care for people suffering from depression: the need for comprehensive education. AB - In the years 1983-1984, the Swedish Committee for Prevention and Treatment of Depressions (PTD) offered an educational programme to all general practioners (GPs) on the Swedish island of Gotland. This education has led to a significant decrease in inpatient care, morbidity, mortality and costs incurred by depressive illness on the island. Unspecific medication decreased and specific antidepressive medication increased. Recent scrutiny of all suicides on Gotland during the 1980s showed that the overall decrease due to the educational programme was mainly in those committed by females with recognized major depression who had been in contact with GPs. This was expected. However, the number of male suicides was almost unaffected by the educational programme nor by the GPs' improved ability to diagnose and treat depressions. We believe that male depressive suicidants may not be reached by the medical health care system. This might be due to mens' alexithymic incapacity to ask for help and/or their atypical depressive symptoms, manifested by aggressive or abusive behaviour that leads to rejection or mis-diagnosis in the health care system. Consequently, there is underdiagnosis and undertreatment of male depressions which may explain the paradoxical fact that men in Sweden and elsewhere are only half as often depressed but commit suicides up to five times more often than females in Sweden. New sex-specific diagnostical and therapeutical tools as well as sex specific research and comprehensive educational programs concerning depression and suicidality are needed. PMID- 10468332 TI - On what evidence should education concerning depression be based? AB - Disease depresses. As a consequence depressed mood is almost ubiquitous in medicine. This is as true for physical disorders as it is for mental disorders. Depressed mood, moreover, seldom comes alone but is generally accompanied by other troublesome phenomena. In other words, the transition from distress towards depression is a gradual one, at least from the clinical point of view. One could therefore rightfully assume, that most physicians will be well versed in diagnosing and treating depression. Regrettably that is not the case. Depression is an under-diagnosed and under-treated disorder. Hence sophisticated educational programs to enhance the diagnostic and therapeutic skills of those frequently encountering depressive and depressed patients are much in need. The Dep Relief program is a CD-ROM program well suited for this purpose. It is wide-ranging, evidence-based, easily adaptable to various audiences and scrutinized by a panel of international experts. In short, it is an important tool to disseminate up-to date information about mood disorders. PMID- 10468333 TI - Sharing the responsibility for improved care: the example of the Lundbeck International Psychiatric Institute. AB - The Lundbeck International Psychiatric Institute was founded as an independent forum for discussions by international psychiatrists on the treatment of mood disorders and schizophrenia with the aim to improve the treatment of patients suffering from CNS-diseases. The Institute has presented 18 schizophrenia seminars until the end of 1998. These seminars address various aspects of the treatment plan of schizophrenia, including psychopharmacology, psychoeducation and quality management. All the discussions are focused on evidenced-based psychiatry and international treatment guidelines. In this paper the data collected from participants of the schizophrenia seminars are presented. These data are comparisons of the psychiatrists' treatment habits and knowledge about schizophrenia before the seminars and 2 weeks and 6 months after attending the seminars. The results presented include changes in the participants' treatment habits in relation to optimal doses for antipsychotics during the acute treatment phase, minimum doses during the maintenance phase, duration of treatment in the maintenance phase, and use of classic versus new generation antipsychotics. The Lundbeck Institute hopes to improve the treatment outcome for patients with CNS diseases. The schizophrenia seminars seem to be changing psychiatrists' treatment habits and their knowledge about the disease. PMID- 10468334 TI - Serial plasma fibronectin levels in pre-eclamptic and normotensive women. AB - OBJECTIVE: Endothelial cell damage has been put forward as an underlying factor for development of pre-eclampsia. This study was carried out to see if fibronectin, which is a marker of endothelial damage, could be used as a marker of pre-eclampsia. METHODS: A longitudinal study was conducted on 100 normotensive primigravidae registered before 20 weeks of gestation. These subjects were followed until delivery and three blood samples were collected, first at registration, i.e. before 20 weeks, second around 28 weeks and third at 36 weeks or later till delivery. Fibronectin levels were assayed by ELISA and women observed for any signs of pre-eclampsia. RESULTS: Fourteen subjects developed pre eclampsia. Fibronectin levels were observed to rise as pregnancy advanced but the rise was significantly higher in subjects who developed pre-eclampsia. The fibronectin levels were also significantly higher in these 14 subjects even in the first sample, i.e. before 20 weeks of gestation when compared with normotensive subjects (P < 0.01). CONCLUSIONS: Fibronectin levels could be used as an early valuable biomarker for the development of pre-eclampsia. PMID- 10468335 TI - Customised gravidogram and fetal growth chart in a South African population. AB - OBJECTIVE: To investigate whether gravidograms and fetal growth curves should be customized, i.e. tailor made for a specific ethnic group. METHODS: A cross sectional study of 800 pregnant Xhosa women attending antenatal care in the Eastern Cape Province (South Africa). The data measured was used to compare the correlation between the symphysis-fundus height measurement (SFHM) and the sonographic estimate of gestational age (SEGA), as well as the intra-uterine fetal growth curve with existing gravidograms and fetal growth charts. RESULTS: There was a good correlation between the SFHM and the SEGA: r = 0.91 (P < 0.0001). The correlation between the SFHM and the sonographic estimate of fetal weight (SEFW) yielded a correlation coefficient (r) of 0.40 (P < 0.0001). The correlation between the SEFW and SEGA resulted in a r = 0.97 (P < 0.0001). The customized gravidogram and fetal growth chart followed the general trend seen in their Caucasian equivalents. CONCLUSION: The comparison of our customized gravidogram and intra-uterine fetal growth curve with similar charts established in Caucasians does not show any significant difference. PMID- 10468336 TI - Preoperative ultrasound assessment of adnexal masses in premenopausal women. AB - OBJECTIVES: This study was undertaken to determine the accuracy of preoperative transvaginal ultrasound in predicting ovarian histopathology in premenopausal women. METHODS: A retrospective cohort study was performed from reproductive-age women that underwent removal of an ovarian cyst between May 1990 and April 1998. RESULTS: Of the 120 consecutive patients identified, 19 did not meet inclusion criteria. Pathologic diagnosis confirmed endometriomas (n = 55), mature teratoma (n = 25), serous cystadenoma (n = 18), persistent corpus luteum (n = 7) and other (n = 5). The primary preoperative transvaginal ultrasound impression was accurate in most cases (endometrioma: 85%, mature teratoma: 86%, serous cystadenoma: 80%). CONCLUSION: Endometriomas and mature cystic teratomas can be predicted with a moderate degree of accuracy by preoperative ultrasound. PMID- 10468337 TI - Vaginal fluid pH as a screening test for vaginitis. AB - OBJECTIVE: To assess how effective the pH test can detect infectious vaginitis. METHODS: Ambulatory gynecological patients attending the gynecological out patient department of Srinagarind Hospital from May 1 to July 31, 1997 were assessed for vaginitis by history, overall physical examination and vaginal examination. Specimens were collected for microbiological examination and measurement of pH level. RESULTS: Among 422 women recruited, a vaginal fluid pH level greater than 4.5 was found in 149 (35.3%) cases. The vaginal fluid pH as a screening test for infectious vaginitis showed a sensitivity of 49.7% (95% C.I.: 42.6-56.9). When using vaginal fluid pH combined with clinical symptoms and signs to screen for vaginitis the sensitivity was 67.5% (95% C.I.: 60.4-73.9). If a pH test was used to screen for BV, its sensitivity was 73.4% (95% C.I.; 60.7-83.3). Using a pH test in combination with clinical symptoms and signs of vaginitis to screen for BV, its sensitivity was 81.3% (95% C.I.: 69.2-89.5). CONCLUSION: Vaginal fluid pH combined with clinical symptoms and signs had a considerably high sensitivity to screen for BV. PMID- 10468338 TI - Genetic analysis of familial and multiple malignancies of endometrial cancer. AB - The presence of the positive replication errors (RER) phenotype in familial and multiple primary malignancies of endometrial cancer, and its association with a poor prognosis was examined. We analyzed 40 endometrial cancers for RER. Eight endometrial cancers with the RER(+) phenotype at multiple microsatellite loci were detected. The presence of the RER(+) phenotype was higher than in non familial malignancies. None of the eight cases with the RER(+) phenotype involved multiple primary malignancies; however these patients had shorter survival times. In this study, we suggest that RER examination in endometrial cancer may be useful for establishing a diagnosis of a familial malignancy, and for predicting a poor prognosis. PMID- 10468339 TI - Prognostic factors relating to survival in uterine endometrioid carcinoma. AB - OBJECTIVE: Epidemiologic and clinicohistopathologic prognostic factors of uterine endometrioid carcinomas were analyzed. The association of estrogen related factors, focused on adenomyosis in the prognosis of endometrioid carcinomas was also examined. METHODS: Risk factors of surgically treated 286 patients with endometrioid carcinoma (Stage I-III) were statistically analyzed. RESULTS: Overall a recurrence-free 5-year survival rate was 81% (Stage I, 94%, Stage II, 71% and Stage III, 40%). Significant prognostic factors were lymph node metastases (P = 0.0035) and serosal/parametrial invasion (P = 0.014) by multivariate analysis. Endometrioid carcinomas with co-existing adenomyosis tend to be associated with endometrial hyperplasia (P = 0.04, Fisher's exact test), diagnosed in less invasive status (myometrial invasion, P = 0.004 and serosal/parametrial invasion, P = 0.006) and therefore have a favorable prognosis (P = 0.01, log rank test). CONCLUSIONS: A favorable prognosis of endometrioid carcinomas with co-existing estrogen related factors (adenomyosis and endometrial hyperplasia) was suggested. PMID- 10468340 TI - The effect of maternal hypothermia on the fetal heart rate. AB - Fetal bradycardia is a recognized response to maternal hypothermia but has not previously been reported in conjunction with diabetes. A 30-year-old insulin dependent diabetic was admitted at 35 weeks gestation for control of her diabetes. She developed maternal hypothermia and hypoglycemia and the fetal heart rate fell to 100 beats per minute (b.p.m.). However, the fetal heart rate gradually returned to normal after rewarming the patient. PMID- 10468341 TI - First trimester sonographic diagnosis of holoprosencephaly. AB - OBJECTIVE: To describe our experience with sonographic diagnosis of fetal holoprosencephaly in first trimester. SUBJECTS: A total of three fetuses with early prenatal diagnosis of holoprosencephaly were sonographically evaluated and followed up. RESULTS: The study revealed that all showed monoventricular cavity, fused thalami, no falx and cavum septum pellucidum. All of them were correctly diagnosed sonographically in the first trimester. Extracranial anomalies had also been identified in all three fetuses and all of them had facial abnormalities. Cytogenetic studies were successfully carried out in only one case. No polyhydramnios was demonstrated in all cases. CONCLUSION: This small series indicates that holoprosencephaly can be diagnosed in the first trimester. The most valuable clue to the diagnosis is the demonstration of the single ventricle. PMID- 10468342 TI - Seroprevalence of syphilis, human immunodeficiency virus type-1, and hepatitis B virus infections among pregnant women in Turkey. PMID- 10468343 TI - Major postpartum hemorrhage as an initial presentation of acute myeloid leukemia. PMID- 10468344 TI - Epidemiology of molar pregnancies in Northern Ireland. PMID- 10468345 TI - Ectopic pregnancy in Ilorin, Nigeria. PMID- 10468346 TI - Sheehan's syndrome in Saudi Arabia. PMID- 10468347 TI - Human immunodeficiency virus infection in pregnant women in Nigeria. PMID- 10468348 TI - Endometrial hyperplasia co-existing with cervical polyps. PMID- 10468349 TI - Urethral prolapse in a west African hospital. PMID- 10468350 TI - Gonadotropin-releasing hormone-agonist as a neoadjuvant therapy for Sertoli Leydig cell tumors of the ovary. PMID- 10468351 TI - Estrogen and lipid profile in patients with endometriosis treated by GnRH agonist. PMID- 10468352 TI - Bone mineral density in natural and surgically-induced menopause. PMID- 10468354 TI - ACOG practice bulletin. Vaginal birth after previous cesarean delivery. Number 5, July 1999 (replaces practice bulletin number 2, October 1998). Clinical management guidelines for obstetrician-gynecologists. American College of Obstetricians and Gynecologists. PMID- 10468353 TI - ACOG committee opinion. Delineation of obstetric-gynecologic primary care practice. Number 218, June 1999. Committee on Primary Care. American College of Obstetricians and Gynecologists. PMID- 10468355 TI - A steady flow analysis on the stented and non-stented sidewall aneurysm models. AB - As part of a general investigation on the effects of blood flow patterns in sidewall aneurysm, in vitro steady flow studies on rigid aneurysm models have been conducted using Particle Image Velocimetry over a range of Reynolds number from 200 to 1600. Above Reynolds number 700, one large recirculating vortex would be formed, occupying the entire aneurysmal pouch. The centre of the vortex is located at region near to the distal neck. A pair of counter rotating vortices would however be formed at Reynolds numbers below 700. For all the aneurysm models considered, the vortex strength, in general, is stronger at higher Reynolds numbers but lower at larger aneurysm size. Maximum strength of the vortex is about 15% of the bulk mean velocity in the upstream parent tube. Estimates of the wall shear stresses are derived from the near wall velocity measurements. Highest level of wall shear stresses always appears at the distal neck of the aneurysmal pouch. Stents and springs of different porosity have been used to dampen the flow movement inside the aneurysm so as to induce the possible formation of thrombosis. It is found that the flow movement inside the aneurysmal pouch can be suppressed to less than 5% of the bulk mean velocity by both devices. Furthermore, regions of high wall shear stresses at the distal neck could also be suppressed by almost 90%. The present results would be useful for further improvements in stent (or spring) technology. PMID- 10468356 TI - Effects of local skull inhomogeneities on EEG source estimation. AB - The accuracy of the head model affects the solutions of the EEG inverse problems. If a simple three-sphere model and standard conductivity values for brain, skull and scalp regions are used, significant errors may occur in the dipole localisation. One of the most sensitive head model parameters is the conductivity of the skull. A realistic three-dimensional finite-element model provides a method to study the effect of inhomogeneities of the skull on the solutions of EEG inverse problems. In this paper the effect of a local skull conductivity inhomogeneity on source estimation accuracy is analyzed by computer simulations for different numbers of electrodes. It is shown that if the inhomogeneity of the skull conductivity is not taken into account, localisation errors of approximately 1 cm can be encountered in the equivalent current dipole estimation. This modelling error introduces a bias to the solution which cannot be compensated by increasing the number of electrodes. PMID- 10468357 TI - Pulse wave attenuation measurement by linear and nonlinear methods in nonlinearly elastic tubes. AB - Reasons for the continuing difficulty in making definitive measurements of pulse wave attenuation in elastic tubes and arteries in the presence of reflections are sought. The measurement techniques available were re-examined in elastic tubes mimicking the arterial compliance nonlinearity, under conditions of strong reflection. The pulse was of physiological shape, and two different pulse amplitudes in the physiological range were used. Measurements of pressure, flow rate and diameter pulsation allowed the deployment of four of the classical linear methods of analysis. In addition, a method of separating the forward- and backward-travelling waves that does not require linearising assumptions was used, and the attenuation in the forward and reverse directions was calculated from the resulting waveforms. Overall, the results obtained here suggest that a fully satisfactory way of measuring arterial attenuation has yet to be devised. The classical linear methods all provided comparable attenuation estimates in terms of average value and degree of scatter across frequency. Increased scatter was generally found at the higher pulse amplitude. When the forward waveforms from the separation were similarly compared in terms of frequency components, the average value at energetic harmonics was similar to both the value indicated by the linear methods and the values predicted from linear theory on the basis of estimated viscous and viscoelastic parameter data. The backward waveforms indicated a physically unreasonable result, attributed as the expression for this technique of the same difficulties that normally manifest in scatter. Data in the literature suggesting that one of the classical methods, the three-point, systematically over-estimates attenuation were not supported, but it was confirmed that this method becomes prone to negative attenuation estimates at low harmonics as pulse amplitude increases. Although the goal of definitive attenuation measurement remains elusive, the task provides a sensitive tool for the examination of the effect of nonlinearities in the arterial system. PMID- 10468358 TI - Biorthogonal wavelet transforms for ECG parameters estimation. AB - The parameters of various morphologies of ECG waveform are basic in characterizing them as normal or otherwise. The use of multiscale analysis, through biorthogonal wavelets presented in this paper, appears very promising for such a characterization. This is on account of the fact that various morphologies are excited better at different scales. From these different scales, amplitudes, durations and various segments, widths can be determined more accurately. Simulation studies, with real ECG data, have shown that even when the signal-to noise ratios are poor, the proposed technique can be used to accurately estimate the said parameters. PMID- 10468359 TI - Viscoelasticity modulates resonance in the terminal aortic circulation. AB - We used an inertance-viscoelastic windkessel model (IVW) to interpret aortic impedance patterns as seen in the terminal aortic circulation of the dog, and to explain evident oscillatory phenomena in flow measurements. This IVW model consists of an inertance, L, connected in series with a viscoelastic windkessel (VW) where the peripheral resistance, Rp, is connected in parallel with a Voigt cell (a resistor, Rd, in series with a capacitor, C) to account for viscoelasticity. Pressure and flow measurements were taken from the terminal aorta, just downstream of the origin of renal arteries, in three anaesthetised open-chest dogs, under a variety of haemodynamic conditions induced by administering a vasoconstrictor agent (methoxamine) and a vasodilator (sodium nitroprusside). Mean pressure ranged from 40 to 140 mm Hg. The resistance Rp was calculated as the ratio of mean pressure to mean flow. Parameters L, C and Rd were estimated by fitting measured to model predicted flow waves. We found that prominent oscillations observed in flow waves, from midsystole to diastole, are related to resonance that occurs at a frequency, f(o), where reactance of inertance of blood motion matches the reactance of arterial compliance. Estimates of f(o) increased from 2.4 to 10 Hz with increasing pressure and showed a correlation with values of static elastic moduli plotted against mean pressure of dogs' peripheral arteries previously reported by others. Viscous losses, Rd, of arterial wall motion limited the amplitude of resonance peak. We conclude that viscoelasticity, rather than pure elasticity, is a key issue to interpret terminal aortic impedance as it relates to resonance. PMID- 10468360 TI - Computational fluid dynamic simulations of cavopulmonary connections with an extracardiac lateral conduit. AB - Complex congenital heart defects due to the absence of a ventricular chamber can often be treated by the Fontan surgical procedure. The objective of this work was to quantify the haemodynamics in the Fontan operation (cavopulmonary connection) with extracardiac lateral conduit. Four different models based on the finite element method were constructed with different lengths of inferior anastomosis (range 18-25 mm) and inclinations of the conduit (33 and 47.5 degrees). Mass conservation and Navier-Stokes equations were solved by means of the FIDAP code, based on the finite element method. The left-to-right pulmonary flow ratio and percentage inferior caval blood to the left lung were the highest with the smallest anastomosis and highest inclination: 1.35 and 83.26%, respectively. Dissipated power percentage was higher with the largest anastomosis than with the smallest (19.4 vs 15.8%). It was concluded that, when performing a total cavopulmonary connection, an extracardiac lateral conduit: (i) diverts more flow to the left lung, and (ii) shows higher energy losses when compared with a connection with intra-atrial tunnel. This study could be useful to evaluate the incidence of pulmonary arteriovenous malformations. PMID- 10468361 TI - Procedure for the semi-automatic detection of gastro-oesophageal reflux patterns in intraluminal impedance measurements in infants. AB - The diagnosis of gastro-oesophageal reflux (GOR) is of great interest for paediatric gastroenterologists. pH monitoring is the commonly used procedure for GOR diagnosis but a major amount of postprandial GOR is missed due to the mostly non-acidic gastric contents in infants. The multiple intraluminal impedance technique is based on the recording of the impedance changes during bolus transport inside the oesophagus. It is the first method which allows the pH independent, long-term registration of GOR. The use of the impedance technology in clinical practice has been limited so far by the time-consuming, visual evaluation of the impedance traces. The new approach of a semi-automatic analysis of the impedance measurements allows the automated detection of reflux patterns. It is based on event marking and an optimised feature description of the impedance traces combined with a fuzzy system for pattern recognition. The classifier is developed and tested on 50 investigations in infants. Compared to the comprehensive, multiple visual evaluation the achieved precision is 75% sensitivity and 48% positive prediction. In comparison to a single visual evaluation the analysis of the automatically proposed patterns corresponds to a 96% reduction of the evaluation time with no loss of precision. Thus the applicability of the impedance technology is enhanced significantly. A combined measurement of pH and impedance gives evidence about the occurrence of GOR, its pH and the acidic exposure of the oesophagus. PMID- 10468362 TI - Hyper-elastic model analysis of anterior cruciate ligament. Shunji Hirokawa and Reiji Tsuruno, Medical Engineering & Physics 1997;19:637-51. PMID- 10468363 TI - Common effects of emotional valence, arousal and attention on neural activation during visual processing of pictures. AB - Emotion and attention heighten sensitivity to visual cues. How neural activation patterns associated with emotion change as a function of the availability of attentional resources is unknown. We used positron emission tomography (PET) and 15O-water to measure brain activity in male volunteers while they viewed emotional picture sets that could be classified according to valence or arousal. Subjects simultaneously performed a distraction task that manipulated the availability of attentional resources. Twelve scan conditions were generated in a 3 x 2 x 2 factorial design involving three levels of valence (pleasant, unpleasant and neutral), two levels of arousal and two levels of attention (low and high distraction). Extrastriate visual cortical and anterior temporal areas were independently activated by emotional valence, arousal and attention. Common areas of activation derived from a conjunction analysis of these separate activations revealed extensive areas of activation in extrastriate visual cortex with a focus in right BA18 (12, -88, -2) (Z=5.73, P < 0.001 corrected) and right anterior temporal cortex BA38 (42, 14, -30) (Z=4.03, P < 0.05 corrected). These findings support an hypothesis that emotion and attention modulate both early and late stages of visual processing. PMID- 10468364 TI - Does parietal cortex contribute to feature binding? AB - We studied the involvement of the right parietal cortex in visual conjunction search, where two features are present in the array and spatial attention and feature binding is required, and in subset search, where two features are also present but only one of them is needed in order to group stimuli together (the subset) and allow parallel processing without the need for feature binding. Six patients with right parietal lobe lesions, six age-matched controls, and three control patients with left parietal lesions were tested on these two tasks. Patients with right parietal lesions were significantly slower than normal controls in the conjunction task, especially for target-absent trials. In the subset condition, neither normal control subjects nor patients with left parietal damage showed a difference between target-present and target-absent trials whereas right parietal patients showed a significant difference between target present and target-absent responses. The results suggest a role for the right parietal cortex in shifting attention to the next stimulus once binding of features has taken place or selecting spatial areas containing the desired feature in a subset search, but that parietal cortex is not required for binding the features of the object. PMID- 10468365 TI - Dissociation of attentional processes in patients with focal frontal and posterior lesions. AB - A location-based ('select-what, respond-where') priming task was used to examine three measures of selective attention (interference (INT), negative priming (NP), and inhibition of return (IOR)) as a function of focal brain pathology and the complexity of target selection. Control subjects showed different patterns of performance for the three attentional measures as a function of complexity, suggesting some independence among INT, NP, and IOR. Brain-damaged subjects showed significant response slowing, as well as a number of lesion-specific attentional abnormalities. Right frontal (including bifrontal) damage resulted in proportionally increased interference related to task complexity. Left posterior damage increased IOR in the most complex task, while left frontal damage reversed the control pattern of IOR as a function of complexity. Right hemisphere (right posterior and right frontal damage) pathology resulted in a virtual loss of negative priming at all levels of task complexity; left and bifrontal damage resulted in diminished NP only related to increases in the complexity of selection. INT, NP, and IOR are mediated by different brain regions and their expression can be modulated by the complexity of the selection task. PMID- 10468366 TI - Imagined transformations of bodies: an fMRI investigation. AB - A number of spatial reasoning problems can be solved by performing an imagined transformation of one's egocentric perspective. A series of experiments were carried out to characterize this process behaviorally and in terms of its brain basis, using functional magnetic resonance imaging (tMRI). In a task contrast designed to isolate egocentric perspective transformations, participants were slower to make left-right judgments about a human figure from the figure's perspective than from their own. This transformation led to increased cortical activity around the left parietal-temporal-occipital junction, as well as in other areas including left frontal cortex. In a second task contrast comparing judgments about inverted figures to judgments about upright figures (always from the figure's perspective), participants were slower to make left-right judgments about inverted figures than upright ones. This transformation led to activation in posterior areas near those active in the first experiment, but weaker in the left hemisphere and stronger in the right, and also to substantial left frontal activation. Together, the data support the specialization of areas near the parietal-temporal-occipital junction for egocentric perspective transformations. These results are also suggestive of a dissociation between egocentric perspective transformations and object-based spatial transformations such as mental rotation. PMID- 10468367 TI - Hemispheric dominance and gender in the perception of an illusion. AB - Perception of geometric illusions is a visuo-spatial process. As such processes often have been found to be predominantly the domain of the right hemisphere, this hemisphere may be expected to perceive such illusions more readilly than the left hemisphere. Using the herringbone illusion in a reaction-test paradigm, we found that in right-handed males the right hemisphere was significantly more often deceived than the left, whereas no significant hemispheric difference was observed in females. This is the first demonstration of gender differences in the lateralized perception of an illusion. PMID- 10468368 TI - Lexical and conceptual components of stem completion priming in patients with Alzheimer's disease. AB - This study evaluated the hypothesis of dissociation between normal lexical but deficient conceptual repetition priming in patients with Alzheimer's disease (AD). For this purpose, we administered to patients with AD and age-matched normal controls the Stem Completion task. In Experiment 1, the level of word processing during study was manipulated by requiring subjects to count vowels (graphemic condition) or generate meanings (semantic condition) of target words. In Experiment 2, the presentation modality was varied during the study to obtain an intramodal and crossmodal repetition priming. Probably due to a floor effect of performance in the graphemic condition, in Experiment 1, AD patients exhibited lower priming than normal controls for the semantically processed words but comparable priming for the graphemically processed ones. In contrast, in Experiment 2, AD patients were poorly primed both in the intra- and crossmodal conditions. Results question the hypothesis of a lexical/conceptual dissociation in the repetition priming exhibited by AD patients and call for other explicative hypotheses of the dissociation between normal and deficient forms of repetition priming in degenerative dementia. PMID- 10468369 TI - Interval timing performance in temporal lobe epilepsy: differences between patients with left and right hemisphere foci. AB - This experiment examined interval timing performance on a temporal reproduction procedure and a temporal discrimination (interval bisection) procedure in 19 patients with temporal lobe epilepsy (ten with a left-hemisphere focus [LTE group] and nine with a right- hemisphere focus [RTE group]), and 14 normal control subjects. In the temporal reproduction task, subjects were required to reproduce the durations of visual stimuli (0.5, 1, 2, 4, 8 s). In the temporal discrimination task, subjects were required to classify the visual stimuli as either 'short' or 'long'. Following exposure to the two standard durations (1 and 2 s), 'probe' trials were introduced in which the stimulus was presented for durations intermediate between the two standard durations. Psychophysical functions were derived from both timing tasks for each individual subject, as well as for the group mean data. The results showed that, compared to the normal subjects, the RTE group's timing ability was significantly compromized, as reflected by larger Weber fractions in both timing tasks. The LTE group's Weber fractions did not differ significantly from those of the control group; however they showed a leftward shift (i.e. a shorter bisection point) of the psychophysical function under the temporal discrimination task. The results suggest that the right and left hemispheres may play different roles in regulating interval timing performance. PMID- 10468370 TI - Differences in semantic category priming in the left and right cerebral hemispheres under automatic and controlled processing conditions. AB - The contribution of each cerebral hemisphere to the generation of semantic category meanings at automatic and strategic levels of processing was investigated in a priming experiment where prime and target words were independently projected to the left or right visual fields (LVF or RVF). Non associated category exemplars were employed as related pairs in a lexical decision task and presented in two experimental conditions. The first condition was designed to elicit automatic processing, so related pairs comprised 20% of the positive set, stimulus pairs were temporally separated by a stimulus onset asynchrony (SOA) of 250 ms, and there was no allusion to the presence of related pairs in the instructions to subjects. The second condition, designed to invoke controlled processing, incorporated a relatedness proportion of 50%, stimulus pairs separated by an SOA of 750 ms, and instructions which informed subjects of the presence and use of category exemplar pairs in the stimulus set. In the first condition, a prime directed to either visual field facilitated responses to categorically related targets subsequently projected to the RVF, while in the second condition a prime directed to either visual field facilitated responses to related targets projected to the LVF. The facilitation effects obtained in both conditions appeared to reflect automatic processes, while strategic processes were invoked in the left, but not the right hemisphere in the second condition. The results suggest that both hemispheres have automatic access to semantic category meanings, although the timecourse of activation of semantic category meanings is slower in the right hemisphere than in the left. PMID- 10468371 TI - Tactile morphagnosia secondary to spatial deficits. AB - A 73-year old man showed visual and tactile agnosia following bilateral haemorrhagic stroke. Tactile agnosia was present in both hands, as shown by his impaired recognition of objects, geometrical shapes, letters and nonsense shapes. Basic somatosensory functions and the appreciation of substance qualities (hylognosis) were preserved. The patient's inability to identify the stimulus shape (morphagnosia) was associated with a striking impairment in detecting the orientation of a line or a rod in two- and three-dimensional space. This spatial deficit was thought to underlie morphagnosia, since in the tactile modality form recognition is built upon the integration of the successive changes of orientation in space made by the hand as it explores the stimulus. Indirect support for this hypothesis was provided by the location of the lesions, which could not account for the severe impairment of both hands. Only those located in the right hemisphere encroached upon the posterior parietal cortex, which is the region assumed to be specialised in shape recognition. The left hemisphere damage spared the corresponding area and could not, therefore, be held responsible for the right hand tactile agnosia. We submit that tactile agnosia can result from the disruption of two discrete mechanisms and has different features. It may arise from a parietal lesion damaging the high level processing of somatosensory information that culminates in the structured description of the object. In this case, tactile recognition is impaired in the hand contralateral to the side of the lesion. Alternatively, it may be caused by a profound derangement of spatial skills, particularly those involved in detecting the orientation in space of lines, segments and complex patterns. This deficit results in morphagnosia, which affects both hands to the same degree. PMID- 10468372 TI - Introduction: Alexander Luria's continuing influence on worldwide neuropsychology. AB - As an introduction to this special issue, this article provides an overview of the worldwide influence of the work of Alexander R. Luria, a noted Russian neuropsychologist. Major themes and issues that he studied are reviewed, and the reasons for his strong worldwide influence are discussed. An overview of subsequent articles in this issue is provided. PMID- 10468373 TI - Outline for the neuropsychological examination of patients with local brain lesions. PMID- 10468374 TI - Quantitative and qualitative integration of Lurian procedures. AB - This article reviews contemporary Russian research aimed at integrating and combining qualitative and quantitative evaluation approaches using Luria's procedures of neuropsychological assessment. A scoring system for rating and evaluating cognitive disturbances in different functional areas is described. The advantages of application of this system are discussed, using as examples the neuropsychological follow-up of neurosurgical patients, as well as neuropsychological research into the cognitive disturbances in patients with Parkinsonism and in children who are mentally retarded. The effectiveness of these updates to Luria's approach is demonstrated. PMID- 10468375 TI - Russian neuropsychology after Luria. AB - It is now more than 20 years after Luria's death in 1977. His collaborators, disciples, and followers both in Russia and abroad continue to further develop his work. The development of Russian neuropsychology reflects the universal tendency to replace static neuropsychology, which relates individuals' behavior to fixed cerebral lesions, with dynamic neuropsychology, which analyzes the dynamics of brain-behavior interaction. Three types of Russian studies illustrate the latter approach: (1) neuropsychological follow-up of different nosological groups of patients in the process of medical or psychological treatment, (2) studies of cognitive evolution (developmental neuropsychology), and (3) studies of cognitive involution (neurogeriatrics). All studies focus on cortico subcortical and interhemispheric relationships. Another change in modern Russian neuropsychology consists of combining the qualitative approach with the quantitative one, but the system of rating is based, following Luria's tradition, on the psychological evaluation of each task's structure and the qualitative analysis of the patients' performance and possibilities for its correction. Hence, Luria's creative and comprehensive approach stimulates the further development of neuropsychology in Russia. PMID- 10468376 TI - Luria in Uzbekistan: the vicissitudes of cross-cultural neuropsychology. AB - If the material conditions of culture shape cognitive structures, as Luria and Vygotsky argued, the "extraordinarily deep and rapid restructuring of historical forms" (Luria, 1971, 265) in the Soviet Republics that followed the Bolshevik revolution of 1917 provided a natural laboratory to determine whether processes of modernization changed traditional ways of thinking. This was the purpose of Luria's 1931 expedition to the Soviet Republic of Uzbekistan in central Asia. Luria's initial reports attracted vitriolic criticism because he had allegedly belittled "primitive" Uzbeki culture. The lasting importance of the Uzbek expedition is its emphasis on culture as a determinant of cognitive processes that remains valid to the present: in 1984, Gilbert replicated Luria's field studies in South Africa with near-identical results. Yet current neuropsychology has been slow to recognize the need for culturally sensitive assessment. PMID- 10468377 TI - Some facts from the biography of A. R. Luria. AB - The biographical sketches from Luria's life reveal the capricious political changes and their effect on the development of Soviet neuropsychology. The picture that emerges is of an individual who survived several ideological reprisals. He introduced Vygotsky into the mainstream of psychology, and continued to advance the study of cognitive functions within the context of neuroanatomy of the brain. And yet, he had an open mind judged by his interest in Freud, and welcoming to his clinic pioneers of psychology such as Piaget, Bruner and Pribram. Also included are intimate impressions of a daughter of her father, who like his young mentor, Vygotsky was a near-genius. He was unvanquished by adversities, doggedly working away to extend the frontiers of the science of human mind. PMID- 10468378 TI - Why should we report adverse incidents? PMID- 10468379 TI - An evaluation of adverse incident reporting. AB - To examine the reliability of adverse incident-reporting systems we carried out a retrospective review of the mother and baby case notes from a series of 250 deliveries in each of two London obstetric units. Notes were screened for the presence of adverse incidents defined by lists of incidents to be reported in accordance with unit protocols. We assessed the percentage of adverse incidents reported by staff to the maternity risk manager at each unit; the percentage of incidents detected by each risk manager, but not reported; and the percentage of incidents identified only by retrospective case note review. A total of 196 adverse incidents was identified from the 500 deliveries. Staff reported 23% of these and the risk managers identified a further 22%. The remaining 55% of incidents were identified only by retrospective case-note review and not known to the risk manager. Staff reported about half the serious incidents (48%), but comparatively few of the moderately serious (24%) or minor ones (15%). The risk managers identified an additional 16% of serious incidents that staff did not report. Drug errors were analysed separately; only two were known to the risk managers and a further 44 were found by case-note review. Incident-reporting systems may produce much potentially valuable information, but seriously underestimate the true level of reportable incidents. Where one risk manager covers an entire trust, rather than a single unit, reporting rates are likely to be very much lower than in the present study. Greater clarity is needed regarding the definition of reportable incidents (including drug errors). Staff should receive continuing education about the purposes and aims of clinical risk management and incident reporting and consideration should be given to designating specific members of staff with responsibility for reporting. PMID- 10468380 TI - Reasons for not reporting adverse incidents: an empirical study. AB - A previous study (Stanhope et al. 1998) established that staff in two obstetric units reported less than a quarter of designated incidents to the units' risk managers. A questionnaire was administered to 42 obstetricians and 156 midwives at the same two obstetric units, exploring the reasons for low rates of reporting. Questions concerned their knowledge of their unit's incident reporting system; whether they would report a series of 10 designated adverse obstetric incidents to the risk manager; and their views on 12 potential reasons for not reporting incidents. Most staff knew about the incident-reporting system in their unit, but almost 30% did not know how to find a list of reportable incidents. Views on the necessity of reporting the 10 designated obstetric incidents varied considerably. For example, 96% of staff stated they would always report a maternal death, whereas less than 40% would report a baby's unexpected admission to the Special Care Baby Unit. Midwives said they were more likely to report incidents than doctors, and junior staff were more likely to report than senior staff. The main reasons for not reporting were fears that junior staff would be blamed, high workload and the belief (even though the incident was designated as reportable) that the circumstances or outcome of a particular case did not warrant a report. Junior doctors felt less supported by their colleagues than senior doctors. Current systems of incident reporting, while providing some valuable information, do not provide a reliable index of the rate of adverse incidents. Recommended measures to increase reliability include clearer definitions of incidents, simplified methods of reporting, designated staff to record incidents and education, feedback and reassurance to staff about the nature and purpose of such systems. PMID- 10468381 TI - Screening for adverse events. AB - Adverse events (AEs) in medical patients are common, costly, and often preventable. Development of quality improvement programs to decrease the number and impact of AEs demands effective methods for screening for AEs on a routine basis. Here we describe the impact, types, and potential causes of AEs and review various techniques for identifying AEs. We evaluate the use of generic screening criteria in detail and describe a recent study of the sensitivity and specificity of individual generic screening criteria and combinations of these criteria. In general, the most sensitive screens were the least specific and no small sub-set of screens identified a large percentage of adverse events. Combinations of screens that were limited to administrative data were the least expensive, but none were particularly sensitive, although in practice they might be effective since routine screening is currently rarely done. As computer systems increase in sophistication sensitivity will improve. We also discuss recent studies that suggest that programs that screen for and identify AEs can be useful in reducing AE rates. While tools for identifying AEs have strengths and weaknesses, they can play an important role in organizations' quality improvement portfolios. PMID- 10468382 TI - A new learning environment: combining clinical research with quality improvement. AB - The emphasis provided by quality improvement strategies on performance measurement and evaluation often results in our understanding of processes of care and, perhaps, better outcomes. There are different references for process evaluation: external peers, regional profiles of performance or a trending of one's own performance patterns. This paper proposes a methodology that enables learning from the daily practice of medicine by comparing alternative care processes and outcomes. Since it is estimated that 15-20% of medical practices are based on rigorous scientific data establishing their effectiveness, we have much to learn. We propose to learn from our daily practice by combining clinical research methods with quality improvement tools. The products comprise modified clinical trial and case-control studies. In a modified clinical trial, we would use a practice guideline as a control group and modify the guideline to create an experimental group. This method would maintain the internal validity of efficacy research while maintaining the external validity of effectiveness research. In the case-control method, it is possible to quantitate risk for a given outcome and focus improvement effort on factors associated with that outcome. We believe physicians will accept this learning approach because it is a more valid learning method than traditional quality improvement and, unlike randomized clinical trials, learning will occur in the daily practice of medicine. PMID- 10468383 TI - Finding the ethical standard of medical science in the age of the sciences. PMID- 10468384 TI - Assessing the outcomes of total knee replacement. AB - The past few years have seen a growth of interest in outcome measurement in a variety of settings including audit, health care management and commissioning - besides the traditional applications in research work. This paper reports on a study of the outcomes of total knee replacement in an acute hospital where the outcomes were studied as part of an audit process. The outcome measures used included clinical and symptomatic measures as well as generic health status scales. The initial study in one hospital was expanded to include a number of others in the same region and a comparative database of outcomes developed. Examples of the results are shown. The technical measures using knee scores and general health status measure show significant improvement from pre-operatively to 3 months later. This improvement was maintained up to the 1-year follow-up on both measures. Although the information systems for collecting and measuring outcomes has been successful, the ability of such measures to lead to behavioural change has been limited. The problems in using outcome measures are discussed in particular in the context of an audit within hospitals, and for purchasing agencies. PMID- 10468385 TI - Evaluating changes in life expectancy and survival in the elderly. PMID- 10468386 TI - Evaluating the presentation and management of upper respiratory tract infection in primary care clinics in Saudi Arabia: biomedical factors do not govern clinical decision making. PMID- 10468387 TI - Uptake of research findings into clinical practice: a controlled study of the impact of a brief external intervention on the use of corticosteroids in preterm delivery. PMID- 10468388 TI - Screening for diabetic retinopathy: a survey of health authorities during a period of transition. PMID- 10468390 TI - Effective care of acute asthma in the accident and emergency department. PMID- 10468389 TI - Pre-hospital care in Valparaiso--an integrated emergency network within the San Antonio Regional Health Service in Chile. PMID- 10468391 TI - Informal economic activities of public health workers in Uganda: implications for quality and accessibility of care. AB - This paper reports the results of a study in Uganda of the 'informal' economic activities of health workers, defined as those which earn incomes but fall outside official duties and earnings. The study was carried out in 10 sub hospital health facilities of varying size and intended role and used a variety of quantitative and qualitative methods. The paper focuses on those activities which are carried out inside public health facilities and which directly affect quality and accessibility of care. The main strategies in this category were the leakage of drug supply, the informal charging of patients and the mismanagement of revenues raised from the formal charging of patients. Few of the drugs supplied to health units were prescribed and issued in those sites. Most health workers who have the opportunity to do so, levy informal charges. Where formal charges are collected, high levels of leakage occur both at the point of collection and at higher levels of the system. The implications of this situation for the quality and accessibility of services in public health facilities were assessed. Utilisation levels are less than those expected of the smallest rural units and this workload is managed by a handful of the expected staff complement who are available for a fraction of the working week. Even given these few patients, drugs available after leakage were sufficient to cover less than half of those attending in most facilities. Evidence on staff motivation was mixed and better motivation was associated with better performance only in a minority of units. Informal charging was associated with better performance regarding hours worked by health workers and utilisation rates. Drug leakage was associated with worse performance with respect to both of these and unsurprisingly, with drug availability. Short term strategies to effect marginal performance improvements may focus on the substitution of strategies based inside health units (such as informal charging) for those based outside (facilitated by drug leakage). In the long term, only substantially higher funding of the sector can be expected to facilitate major change, but alone will be insufficient. Investment strategies supported by appropriate policy development has to be informed by understanding and monitoring of the 'informal' dimension of health sector activity. PMID- 10468392 TI - Do measures of self-reported morbidity bias the estimation of the determinants of health care utilisation? AB - Most national surveys of health care utilisation capture only self-reported measures of morbidity. If self-reported morbidity is measured with error, then the results of applied work may be misleading. In this paper we propose a model of the relationship between morbidity and health service utilisation which allows for reporting errors and simultaneity. Errors in self-reported morbidity are expressed as a function of person-specific reporting thresholds and recent contact with health services, arising because of better self-evaluation of current health status or a desire to justify consumption of a publicly-provided good. We demonstrate the bias in ignoring the potential problems of reporting errors and simultaneity for a variety of special cases, but in the general case the biases are of ambiguous sign. The empirical nature of these biases is investigated using limiting long-standing illness (LLI) and recent contact with a General Practitioner (GP) in two waves of The UK Health and Lifestyle Survey. Biomedical measures of functioning are used as objective indicators of health status. We find evidence of substantial and significant differences between individuals in reporting thresholds and some evidence that the reporting of LLI may depend on recent visits to a GP. Adjustments for these biases significantly increase the estimated effect of morbidity on utilisation. PMID- 10468393 TI - Does the work environment contribute to excess male mortality? AB - It is sometimes assumed that male jobs are on average more unhealthy than female jobs. The aim of the present study is to examine whether work-related factors contribute to excess male mortality. All Swedish deaths during 1970-80 and 1980 86 were analysed with Poisson regressions--for all individuals and for labour force participants--in order to estimate gender mortality rate ratios for all causes, circulatory diseases and external causes. Results for all men and women, as well as results restricted to those in full-time employment, revealed that no work-related factors contribute to excess male mortality. For the period 1980-86, a more detailed analysis was performed, and work environment exposures were aggregated from a secondary data source. In accordance with previous studies, it was found that men experience unhealthier physical work environments than women and that women experience unhealthier psychosocial work environments than men. Among labour force participants, men's greater exposure to hazardous work contributed to men's excess of external causes of death. This finding however, applied only to part-time workers. In conclusion, when factors such as women's greater likelihood of having a low socioeconomic status, their poor psychosocial work environment (low levels of work control and workplace social support), and the fact that women working full-time are more likely than men to be unmarried, were taken into account, the work environment did not appear to contribute to excess male mortality. It is suggested that the lack of reduction in excess male mortality during the 1970 s could be a consequence of the following: women commonly took jobs with low mortality rates; the rapid increase in the numbers of Swedish women in paid employment probably lowered their mortality; other social, behavioural and bio-genetical factors are more important than work in determining excess male mortality. Causality considerations are also discussed. PMID- 10468394 TI - How stable are people's preferences for giving priority to severely ill patients? AB - BACKGROUND: Previous studies have suggested that people favor allocating resources to severely ill patients even when they benefit less from treatment than do less severely ill patients. This study explores the stability of people's preferences for treating severely ill patients. METHODS: This study surveyed prospective jurors in Philadelphia and asked them to decide how they would allocate scarce health care resources between a severely ill group of patients who would improve a little with treatment and moderately ill patients who would improve considerably with treatment. Subjects were randomized to receive one of six questionnaire versions, which altered the wording of the scenarios and altered whether subjects were given an explicit option of dividing resources evenly between the two groups of patients. RESULTS: Four hundred and seventy nine subjects completed surveys. The preference subjects placed on allocating resources to severely ill patients depended on relatively minor wording changes in the scenarios. In addition, when given the explicit option of dividing resources evenly between the two groups of patients, the majority of subjects chose to do so. CONCLUSION: People's preferences for allocating resources to severely ill patients can be significantly decreased by subtle wording changes in scenarios. However, this study adds to evidence suggesting that many people place priority on allocating resources to severely ill patients, even when they would benefit less from treatment than others. PMID- 10468395 TI - Policy analysis for end-stage renal disease in Jamaica. AB - We developed and applied methods for policy analysis for end-stage renal disease (ESRD) in Jamaica. Our emphasis was on methods useful for situations often found in developing countries, where both resources and data may be limited. Many countries are experimenting with ESRD treatment options, but little analysis has been done regarding how developing countries should approach policy decisions for ESRD. Methods for policy analysis in high-income countries often rely on large data sets that may be unavailable or only partially available in developing countries. We conducted technical analysis applicable in these circumstances and emphasized a process for including a wide range of policymakers and other stakeholders in both quantitative and qualitative aspects of the analysis. Our methods may also be applicable in other developing countries and for other chronic diseases. Our analysis included eight issues: (1) a review of currently available clinical and scientific understanding regarding ESRD; (2) a review of country-specific socioeconomic and clinical issues relevant to ESRD in Jamaica; (3) estimates of the magnitude of the need for treatment in the Jamaican population; (4) comparison of the need with available treatment capacity; (5) cost analysis related to options for expansion of treatment capacity; (6) comparison of costs to government budget resources and other potential sources of financing; (7) development of policy options; and (8) sensitivity testing of policy scenarios and trade-offs with competing priorities. We also identified several key decisions most developing country governments will face in setting health policy for ESRD. These include allocating funds for ESRD, identifying and selecting cost-saving clinical strategies, rationing available treatment capacity, and identifying the appropriate role for public education. PMID- 10468396 TI - Immunization promotion activities: are they effective in encouraging mothers to immunize their children? AB - Mass media communication is an important strategy for increasing parental uptake and to promote community participation when large-scale immunization activities are carried out. In Mexico, the National Vaccination Council (CONAVA) launches three immunization campaigns every year accompanied by three vaccination promotion campaigns. This study was conducted to assess whether communication activities to promote CONAVA's Second National Health Week (SNHW) were effective in providing information to mothers about the importance of immunizing their children under five years of age and in prompting them to seek immunization services. A probability sample of mothers living in the metropolitan area of Mexico City and having at least one child under five years old was selected for the study. Four outcome variables were defined as measuring the impact of the campaign: (1) mothers' knowledge about the SNHW; (2) mothers' comprehension indicating how well they understood the campaign messages (aware, partly aware and unaware); (3) mothers' motivation, i.e. whether or not they sought out immunizations for their children under the age of five and (4) mothers' opinion of how well they liked the messages. A total of 935 mothers were interviewed; 88.2% knew about the SNHW, 64.3% were aware that the campaign aimed to provide immunizations, and most held a favorable opinion about the messages. Among aware mothers, 87.5% of their children received immunizations. In this group 72.1% were prompted by the information in the campaign to seek immunizations for their children while 27.9% had to be personally invited to participate in the campaign. The latter occurred either when health workers or volunteers visited mothers in their homes or by soliciting mothers' participation as they visited or passed by immunization health posts. In the unaware mothers group, 72.7% of their children received immunizations; 62.5% of the mothers took their children because of information they received through the campaign while 37.5% had to be personally invited to immunize their children. Mothers with better socioeconomic status were more aware of the campaign, but a high percentage of them did not seek immunizations, while mothers with middle and lower socioeconomic status were motivated to immunize their children through the campaign. Promotion activities and messages communicated through the mass media were appropriate to inform and motivate mothers to seek immunization services for their children. PMID- 10468397 TI - Risk factors of pain in mammographic screening. AB - Mammography is an important tool in the secondary prevention of breast cancer. However, earlier research has pointed out that an unpleasant experience during a previous mammography can deter women from returning for mammography screening. It is known that mammography can be a painful examination for some women. The research presented in this article focuses on the experience of pain during and after mammography (247 patients). Firstly, the study sets out to determine the extent to which women actually experience pain as a result of mammographic examination. Secondly, it focuses on identifying the factors that determine the risk of a painful experience during the screening procedure. A pain model was developed that takes into account a broad range of potential pain factors, including other than woman-related factors and factors associated with the actual examination. Special attention was paid to the examination context, the mammographic procedure and the screening staff. The research results show that the majority of women experience pain during mammography. With a logistic regression a number of women-related factors, staff-related and procedural factors were found significant in assessing the pain risk during mammography. PMID- 10468398 TI - Inconsistent responses in three preference-elicitation methods for health states. AB - Values for health states obtained from the general population can be used in the development of cost-utility algorithms such as Quality Adjusted Life Years (QALYs) to aid in resource allocation decisions. These values are obtained using methods such as the visual analogue scale, the time trade-off or the standard gamble. However, all of these methods suffer, to some extent, from the problem of inconsistent responses. In this paper, we examine the degree to which three preference-elicitation methods-ranking, VAS and TTO--produce inconsistent responses, the effect of sociodemographic and health variables on the numbers of inconsistencies produced, and the effect of including inconsistent responses on the ordering of health states in EQ-5D VAS and TTO tariffs. Health states valued were a sub-set of 43 health states generated by the EuroQol-5D instrument, which were valued as part of a larger study to obtain a tariff of VAS and TTO values for use with the Spanish version of the EQ-5D. Two types of inconsistency- internal and criterion inconsistency were tested, and both the numbers of inconsistencies produced by each method, and the 'size' of those inconsistencies, i.e. the distance in terms of severity between health states rated inconsistently, were tested. The study showed that although the TTO produces higher numbers of inconsistent responses, the numbers of inconsistent responses in all methods were low, and did not affect rankings in the final tariff of values. Older respondents and those with lower levels of education produced significantly higher numbers of inconsistencies, though health characteristics were not significant. On the ranking and VAS methods, the number of inconsistencies doubled between the two interviewers. Efforts should be made to reduce inconsistencies, particularly among older and lesser educated respondents. Future research should concentrate on determining how inconsistencies might affect the values assigned to health states in the final tariff, and not simply the order of those states. PMID- 10468399 TI - Problems Tamil asylum seekers encounter in accessing health and welfare services in Australia. AB - Over the last decade, western countries have reduced their intake of refugees, even though a substantial number of persons continue to be displaced by war and persecution. At the same time, there has been a substantial increase in the number of asylum seekers who apply for refugee status after entering western countries without resettlement documents. Evidence is accruing that asylum seekers are at high risk to trauma-related psychiatric and physical disorders. Increasing concerns have been raised, therefore, about the difficulties that asylum seekers face in accessing health and welfare services. The present Australian-based volunteer study compared Tamil asylum seekers (n = 62) from Sri Lanka with compatriots (30 refugees; 62 immigrants) on a number of indices relating to difficulties accessing medical, counselling and welfare services. The majority of asylum seekers (>60%) reported serious difficulties accessing medical and dental services and a sizeable minority reported problems obtaining assistance with welfare (40%), counselling (34%), and charity (23%). Difficulties accessing medical and dental services consistently exceeded those reported by refugees and immigrants. In spite of the inevitable sampling limitations, the data support past research as well as clinical impressions in suggesting that asylum seekers are particularly disadvantaged in accessing health care services. PMID- 10468400 TI - Role conflict and rapid socio-economic change: breastfeeding among employed women in Thailand. AB - Conflicts between women's productive and reproductive roles are intensified by rapid development and social change. Women have a right to offer optimum nutrition to their babies through breastfeeding; they also are entitled to seek gainful employment. For many, furthermore, employment is essential to the economic survival of their families. This article derives from a combined qualitative and quantitative study conducted in Chiang Mai, Thailand. Interviews were carried out with 313 women to investigate the experiences of those who resumed employment within six months after delivery. The findings demonstrate that urban women in the modern workplace face many obstacles in their efforts to maintain lactation while simultaneously undertaking paid work. Current public policies do not address these obstacles effectively, which is of particular concern in today's volatile economic climate. PMID- 10468401 TI - Subjective life expectancy in the US: correspondence to actuarial estimates by age, sex and race. AB - This study maps the relationship between subjective and actuarial life expectancy in a 1995 national sample of 2037 Americans of ages 18-95. Subjective estimates parallel age-specific actuarial ones based on current age-specific mortality rates. However males expect to live about 3 years longer than the actuarial estimate and blacks expect to live about 6 years longer. The apparent optimism remains after adjusting for socioeconomic status and the signs and symptoms of good health. Contrary to economists' rational-expectations hypothesis, young adults do not adjust their life expectancies upward to account for the favorable trends in mortality rates. PMID- 10468402 TI - Mortality and migration in Britain, first results from the British Household Panel Survey. AB - This study investigates the extent to which current geographical variations in mortality are influenced by patterns of migration since birth. It is based on a longitudinal study of migrants which consists of a representative sample of 10264 British residents born after 1890 and enumerated as part of the British Household Panel Study in 1991. Between 1991 and 1996, 527 of the study members died and these deaths were analysed by area of residence at birth and in 1991 at both the regional and local district geographical scales. These were compared with findings from the Office for National Statistics Longitudinal Study. The British Household Panel Survey sample replicates the results of work conducted on the Longitudinal Study which finds that geographical variations in age-sex standardised mortality ratios at the regional scale cannot be attributed to selective migration. However, for the British Household Panel Survey sample, the major geographical variations at district level could be attributed to selective migration. Geographical variations in mortality are not well understood. Restrictions on what it is possible to analyse in the Office for National Statistics Longitudinal Study may have resulted in the underestimation of the importance of local lifetime selective migration in producing the contemporary map of mortality variation across Britain. The British Household Panel Survey is a small, recent, but very flexible study, which can be used to investigate the effects of lifetime migration on mortality patterns for all of Britain. This first report of its results on mortality shows that it produces findings which accord with the much larger Longitudinal Study, but which can be taken further to show that selective migration over the whole life-course at the local level does appear to have significantly altered the geographical pattern of mortality seen in Britain today. PMID- 10468403 TI - DNA aneuploidy in ulcerative colitis and in colorectal carcinoma--a comparative study. PMID- 10468404 TI - Origin of microcells in the human sarcoma cell line HT-1080. AB - The aim of this study was to investigate the development of microcells in the human sarcoma cell line HT-1080 after interference with thiophosphamidum. We found that damaged interphase macrocells located at the projection of the nucleolus may form one or several microcells. The micronuclei of the microcells intensively incorporate the thymidine analogue 5-bromo-2'-deoxyuridine and strongly express argyrophilic nucleolar organiser region proteins. At an early phase of the development, the micronuclei contain fragmented DNA, but in subsequent phases, the micronuclei accumulate polymeric DNA, simultaneously with an increase in their size. After desintegration of the damaged macrocell, the microcells appear in the intercellular space. The microcells can enter mitosis and they strongly express the lung resistance protein. Electron microscopic observations suggest that coiled bodies are involved in the development of the microcells. Since the observed path of microcell formation differs from apoptotic cell fragmentation into apoptotic bodies, we propose a new term for this microcell development: sporosis. We suggest that self-renewal of the tumour stem cells is likely based on sporosis. PMID- 10468405 TI - Human breast carcinomal tissues display distinctive FTIR spectra: implication for the histological characterization of carcinomas. AB - Fourier transform infrared spectroscopic study of human breast normal and carcinomal tissues has been carried out. Some distinctive spectral differences which are mainly due to nucleic acids and proteins are observed between normal and carcinomal tissues. This method of analysis results in nearly 100% diagnostic accuracy of carcinomal tissues from normal tissues. The spectral patterns of well differentiated carcinomal tissues exhibit marked heterogeneity, however that of poorly differentiated carcinomas demonstrate significant similarity. Apocrine, tubular, intraductal and mucinous carcinomas and invasive infiltrating ductal carcinomal tissues can be discriminated based on their characteristic spectra. The spectral differences confirm the possibility of using FTIR as an accurate and rapid technique to distinguish between normal and malignant breast tissues and classify breast carcinomas in different subtypes. PMID- 10468406 TI - Influence of sampling practices on the appearance of DNA image histograms of prostate cells in FNAB samples. AB - Twenty-one fine needle aspiration biopsies (FNAB) of the prostate, diagnostically classified as definitely malignant, were studied. The Papanicolaou or H&E stained samples were destained and then stained for DNA with the Feulgen reaction. DNA cytometry was applied after different sampling rules. The histograms varied according to the sampling rule applied. Because free cells between cell groups were easier to measure than cells in the cell groups, two sampling rules were tested in all samples: (i) cells in the cell groups were measured, and (ii) free cells between cell groups were measured. Abnormal histograms were more common after the sampling rule based on free cells, suggesting that abnormal patterns are best revealed through the free cells in these samples. The conclusions were independent of the applied histogram interpretation method. PMID- 10468407 TI - Heterogeneity of DNA distribution in diploid cells: a new predictive discriminant factor for solid tumour behaviour. AB - Spatial nuclear DNA heterogeneity distribution of Feulgen-stained DNA diploid cells was studied by image cytometry in voided urine of 119 patients without bladder tumour (n = 20) and with initial (n = 23) or previous (n = 76) diagnosed bladder tumour. For each patient, repetitive DNA measurements were performed during 1-4 years of follow up. Only cells of diploid DNA histograms and diploid subpopulations of aneuploid DNA histograms were used for analysis. DNA heterogeneity distribution of these diploid cells was quantified by statistical parameters of each nuclear optical density distribution. Discriminant analysis was performed on three groups of DNA histograms. Group A (n = 44): aneuploid DNA histograms of patients with bladder tumour. Group D (n = 55): 38 diploid DNA histograms of the 20 patients without bladder tumour (subgroup D1) and 17 diploid DNA histograms of patients with a non-recurrent bladder tumour (subgroup D2). Group R (n = 27): diploid DNA histograms of patients with bladder tumour recurrence. No statistically significant discriminant function was found to separate D1 and D2. However, the first canonical discriminant function C1 differentiated diploid cells of diploid DNA histograms (group D and group R) from diploid cell subpopulations of aneuploid DNA histograms (group A). Mean C1 values were 1.06, 0.84 and -1.45 for groups R, D and A, respectively. The second canonical discriminant function C2 differentiated diploid DNA histograms of patients with bladder tumour recurrence (group R) from diploid DNA histograms of patients without bladder tumour or without bladder tumour recurrence (group D). Mean C2 values were 1.78 and -0.76 for groups R and D, respectively. In 95% confidence limit, the rate of rediscrimination using the two first canonical discriminant functions C1 and C2 were 86.4, 74.5 and 74.1% for groups A, D and R, respectively. Percent of "grouped" cases correctly classified was 78.6%. Thus spatial DNA heterogeneity distribution of diploid cells seems to quantitate probable genetic instability as a function of clinical evolution such as tumour recurrence, and suggests the possible presence of aneuploid stemlines in a heterogeneous tumour, even if a diploid DNA histogram is observed in a single sample. From standardized C1 and C2 canonical discriminant function coefficients, a DNA heterogeneity index (2c-HI) is proposed to characterize diploid cells providing a descriptive and predictive discriminant factor for solid tumour behaviour. PMID- 10468408 TI - Diagnosis and prognosis of neuroendocrine tumours of the lung by means of high resolution image analysis. AB - Neuroendocrine tumours (NET) of the lung are divided in subtypes with different malignant potential. The first is the benign or low-grade malignant tumours, well differentiated, called typical carcinoids (TC) and the second is the high-grade malignant tumours, poorly differentiated of small (SCLC) or large cell type (LCLC). Between these tumour types lies the well-differentiated carcinoma with a lower grade of malignancy (WDNEC). In clinical routine it is very important with regard to prognosis to distinguish patients with low malignant potential from those with higher ones. In this study 32 cases of SCLC, 13 of WDNEC and 14 of TC with a follow-up time up to 7 years were collected. Sections 4 microm thick from paraffin embedded tissue were Feulgen stained. By means of high resolution image analysis 100 nuclei per case were randomly gathered to extract morphometric, densitometric and textural quantitative features. To investigate the ploidy status of the tumour the corrected DNA distribution was calculated. Stepwise linear discriminant analysis to differentiate the classes and Cox regression analysis for the survival time analysis were applied. Using chromatin textural and morphometric features in two two-class discriminations, 11 of the 14 TC cases and 8 of the 13 WDNEC cases were correctly classified and 11/13 WDNEC cases and 28/32 SCLC cases, respectively. The WDNEC cases are more similar in chromatin structure to TC than to SCLC. For the survival analysis, only chromatin features were selected to differentiate patients with better and worse prognosis independent of staging and tumour type. PMID- 10468409 TI - Sjogren's syndrome: too many associations, too limited evidence. The enigmatic example of CNS involvement. PMID- 10468410 TI - Primary Sjogren's syndrome with severe central nervous system disease. AB - OBJECTIVE: Central nervous system (CNS) involvement in primary Sjogren's syndrome (pSS) is controversial with regard to frequency, significance, and etiology. METHODS: We describe a young woman with pSS and severe CNS disease and review the literature on the pathophysiology, clinical significance, symptoms, diagnostic examinations, and treatment of CNS disease with concomitant pSS (CNS-SS). RESULTS: Our patient with pSS had a 5-month history of benign lymphadenopathy and myositis, after which she developed severe CNS disease, vasculitic lesions on her hands, and a neurogenic bladder attributable to spinal cord involvement. The diagnosis was based on the clinical picture and the results of a brain magnetic resonance imaging (MRI) scan, electroencephalography (EEG), and cerebrospinal fluid (CSF) analysis. The disease did not respond to corticosteroids, but the administration of cyclophosphamide resulted in recovery. In the literature, the incidence of CNS-SS varies widely, from rare to incidence rates of 20% to 25%. The clinical picture is diverse, ranging from mild cognitive symptoms to fatal cerebrovascular accidents. The pathophysiology of CNS-SS is unclear, specific diagnostic methods are not available, and diagnosis is based on the clinical picture and a combination of examinations. MRI is the most sensitive test and cerebral angiography the most specific. CSF reflects involvement of the leptomeninges, and EEG is nonspecific. There are no controlled studies of the treatment of CNS-SS. Regimens for vasculitis are commonly used. CONCLUSIONS: CNS SS is uncommonly recognized and difficult to diagnose. Increasingly accurate and available diagnostic examinations will yield more information about the association of CNS disease with pSS. PMID- 10468411 TI - Methotrexate and leflunomide: biochemical basis for combination therapy in the treatment of rheumatoid arthritis. AB - OBJECTIVES: Methotrexate is currently one of the most widely prescribed disease modifying antirheumatic drugs (DMARDs) for the treatment of rheumatoid arthritis (RA). Combination therapy of methotrexate with other DMARDs increases the clinical success of low-dose methotrexate treatment. Leflunomide is a new DMARD that may have a high potential for success in combination therapy with methotrexate. This review compares the mode of action of methotrexate and leflunomide and speculates on how this contributes to therapeutic efficacy in RA when these agents are used singly or in combination. METHODS: A literature review of the biochemical mechanisms considered to be the basis for the therapeutic efficacy of methotrexate and leflunomide in treating RA is presented. RESULTS: Low-dose methotrexate inhibits cytokine production, purine biosynthesis, and, in an animal model, causes the release of adenosine, a potent antiinflammatory agent. Leflunomide, through inhibition of de novo pyrimidine biosynthesis, can regulate lymphocyte proliferation. CONCLUSIONS: The biochemical mechanisms underlying the therapeutic efficacy of low-dose methotrexate and leflunomide in the treatment of RA are quite different. The potentially complementary mechanisms of action of these two effective DMARDs should provide a rationale for their use in combination therapy for patients whose condition no longer responds to methotrexate alone. PMID- 10468412 TI - T cells: pathogenic cells and therapeutic targets in rheumatoid arthritis. AB - OBJECTIVES: To provide: 1) a brief review of current thought on the role of T cells in the pathogenesis of rheumatoid arthritis (RA); and 2) To provide an overview of RA therapies directed against T cells. METHODS: The following papers in relevant American and European medical journals were reviewed. Those related to: the role of T cells in the pathogenesis of RA; to biological therapy directed against cell surface markers specific to T cell populations implicated in RA; and to treatment of RA with cyclosporin A and leflunomide, pharmacological agents known to interfere with the T cell response to antigens. RESULTS: Although a variety of cell types are now recognized as contributors to the progressive joint destruction that is a hallmark of RA, T cell activation is still thought to be a central event in the initiation and progression of this disease. As a result, various therapeutic options directed against T cells have been developed. These include biological agents directed against specific populations of activated T cells and pharmacological agents that have specific T cell-modulatory actions. CONCLUSIONS: Use of T cell-directed biological therapies for RA has been disappointing, as a result of both lack of efficacy and serious toxicity. Treatment of RA with pharmacological agents that interfere with antigen-driven T cell proliferation has been more successful. PMID- 10468413 TI - Nail fold capillaroscopy: normal findings in children and adolescents. AB - OBJECTIVES: Capillaroscopy is a simple diagnostic method that permits noninvasive in vivo study of the capillary network. Studies designed to standardize capillary normality in children are limited. This article presents the capillaroscopic findings in healthy children and adolescents, thus making the application of this methodology viable for patients in this age range. MATERIAL AND METHODS: Healthy children were recruited from a private elementary school and junior high school. Nail fold capillaroscopy was performed using a stereomicroscope at 16 times magnification, addressing the following parameters: capillary morphology, capillary enlargement, devascularization, microhemorrhage, and subpapillary venous plexus visibility (PVS). These parameters were related to age, sex, ethnicity, and local periungal conditions. RESULTS: The sample comprised 329 individuals with mean age of 8.2 years. We observed atypical capillary morphology in 118 of the studied cases (36%), mainly bizarre capillaries in 90 (27%), meandering capillaries in 32 (10%), and bushy capillaries in 20 (6%). The enlarged capillary phenomenon was uncommon, being observed in 30 cases (9%). The number of capillaries per millimeter varied from five to nine. Deletion areas were detected in only seven individuals (2%). The subpapillary venous plexus was not visualized in 13 (4%) cases. Younger children presented higher PVS scores and fewer capillaries/mm as compared with older children. PVS scores were lower in males and in nonwhite children. Other variables were not associated with sex or ethnicity. CONCLUSIONS: The normal nail fold capillary network in children resembles that observed in adults with some differences, such as a lower number of loops per millimeter, a higher PVS score, and a higher frequency of atypical loops. This information is important for the diagnostic evaluation of children in the context of autoimmune rheumatic diseases. PMID- 10468414 TI - Rheumatic syndromes: clues to occult neoplasia. AB - OBJECTIVES: Rheumatic disorders associated with cancer include a variety of conditions, most of which have no features distinguishing them from idiopathic rheumatic disorders. It is generally held that an extensive search for occult malignancy in most rheumatic syndromes is not recommended unless accompanied by specific findings suggestive of malignancy. The objective of this review are to identify rheumatic syndromes associated with cancer, to call attention to features that may suggest the presence of a hidden cancer, and to examine the role to additional clinical and laboratory data as clues to the possible neoplastic cause of those syndromes. METHODS: A MEDLINE search of the literature dealing with cancer-associated rheumatic syndromes was conducted. RESULTS: Review of the literature identified significant progress in this area. First, the association of malignancy with certain rheumatic syndromes was convincingly established, such as asymmetric polyarthritis presenting in the elderly with an explosive onset, rheumatoid arthritis with monoclonal gammopathy, Sjogren's syndrome with monoclonality, hypertrophic osteoarthropathy, dermatomyositis, polymyalgia rheumatica with atypical features, Lambert-Eaton myasthenic syndrome, palmar fasciitis and arthritis, eosinophilic fasciitis poorly responsive to corticosteroid therapy, erythema nodosum lasting more than 6 months, and onset of Raynaud's phenomenon or cutaneous leukocytoclastic vasculitis after age 50 years. Second, the list of cancer-associated rheumatic syndromes was extended by including additional entities such as benign edematous polysynovitis, sacroiliitis, adult-onset Still's disease, dermatomyositis sine myositis, systemic sclerosis, Sweet's syndrome, osteomalacia, skeletal hyperostosis, antiphospholipid syndrome, and essential mixed cryoglobulinemia. Third, evidence was provided substantiating that certain long-standing rheumatic syndromes, in particular rheumatoid arthritis, Felty's syndrome, Sjogren's syndrome, dermatomyositis, systemic sclerosis, systemic lupus erythematosus, and temporal arteritis behave like "premalignant conditions." Fourth, it was shown that the recognized tumor markers alpha-fetoprotein, prostate-specific antigen, CA-125, CA 19-9, and CA-3 have low sensitivity and specificity in screening for occult cancer in a population of rheumatic patients, whereas the presence of a monoclonal gammopathy in rheumatoid arthritis and the monoclonal antibody 17-109 in Sjogren's syndrome are reliable signs of malignant transformation. CONCLUSIONS: The presence of specific rheumatic syndromes and certain clinical and laboratory findings may justify a workup for hidden cancer. Studies of the epidemiology of the cancer-associated rheumatic syndromes and evaluation of the validity of aforementioned clues in prospective studies are goals for future investigations. PMID- 10468415 TI - Chronic tophaceous gouty arthritis mimicking rheumatoid arthritis. AB - OBJECTIVES: To analyze the factors which differentiate chronic tophaceous arthritis from rheumatoid arthritis. METHODS: We describe two cases of chronic gouty arthritis masquerading as rheumatoid arthritis. The characteristic features of each of these two conditions and the diagnostic approach are discussed in light of relevant literature. RESULTS: The correct diagnosis was reached by the combination of accurate history taking (family history of gout, alcoholism, previous diuretic therapy and renal stones), guiding clinical features (subcutaneous tophaceous deposits) and specific radiological (assymetrical erosions with sclerotic margins and overlying edges) and laboratory findings (hyperuricemia and hyperuricosuria). It was confirmed by the identification of monosodium urate (MSU) crystals in the synovial and subcutaneous tissues. CONCLUSIONS: Gout and rheumatoid arthritis rarely coexist. Chronic gouty arthritis may mimic rheumatoid arthritis, and vice-versa. Clinical suspicion supplemented by characteristic laboratory, radiological and histologic findings help at reaching an accurate diagnosis. PMID- 10468416 TI - Hormone replacement therapy--with or without added progestin--and which progestin? PMID- 10468417 TI - Childhood leukemia and pesticides. PMID- 10468418 TI - Postmenopausal hormone therapy and risk of cardiovascular disease and hip fracture in a cohort of Swedish women. AB - Postmenopausal estrogen use is associated with a reduced risk of heart disease and hip fracture; in observational studies, different behaviors among hormone users and nonusers may partially explain these results. We examined risk of cardiovascular disease and hip fracture with medium-potency compared with low potency or short-term estrogen use, and the effect of added progestin, among 9,236 women in Uppsala, Sweden, who responded to a mailed questionnaire in 1987 1988. Using population registries, we identified 213 cases of myocardial infarction, 289 strokes, and 114 hip fractures from 1987-1995. We found a reduced risk of myocardial infarction for medium-potency compared with low-potency or short-term estrogen use (relative risk = 0.75, 95% confidence interval (CI) = 0.56-0.99), with a similar decrease in the subgroup that took estrogens with progestin (RR = 0.69, 95% CI = 0.45-0.90). There was no relation of medium potency estrogen to stroke (RR = 0.91, 95% CI = 0.71-1.17, and RR = 0.81, 95% CI = 0.61-1.10 for the subgroup taking progestin), and no effect of duration on either heart disease or stroke. We observed a reduction in hip fractures for medium-potency use (RR = 0.65, 95% CI = 0.45-0.95), and for use of combined estrogen-progestin therapy (RR = 0.64, 95% CI = 0.41-1.00). These data support a decreased risk of heart disease and hip fracture for medium-potency estrogen use alone or with progestin; self-selection to hormone use cannot explain these reductions. PMID- 10468419 TI - Risk of childhood leukemia associated with exposure to pesticides and with gene polymorphisms. AB - We conducted a population-based case-control study of childhood acute lymphoblastic leukemia (ALL) to evaluate the risk posed by reported exposure to pesticides used in and around the home. We compared 491 cases 0-9 years of age to as many controls. We also conducted a case-only study on a subsample of 123 cases to evaluate gene-environment interaction between child genotype and maternal exposure during pregnancy as well as child exposure after birth. We used the polymerase chain reaction (PCR) approach to analyze polymorphisms in CYP1A1, CYP2D6, GSTT1, and GSTM1 genes, which encode enzymes involved in carcinogen metabolism. Indoor use of some insecticides by the owners and pesticide use in the garden and on interior plants, in particular frequent prenatal use, was associated with increased risks up to severalfold in magnitude. Interaction odds ratios were increased among carriers of the CYP1A1m1 and CYP1a1m2 mutations when mother during pregnancy or the child had been exposed to certain indoor insecticides. No such effects were observed in the presence of other tested polymorphisms. PMID- 10468420 TI - Lung cancer and indoor air pollution arising from Chinese-style cooking among nonsmoking women living in Shanghai, China. AB - Associations between indoor air pollution from Chinese-style cooking and lung cancer have been found in several investigations. To provide more detailed estimates of the associations while accounting for key confounding factors, we conducted a population-based, case-control study of lung cancer among nonsmoking women living in Shanghai, the People's Republic of China. Five hundred four incident, primary lung cancer cases diagnosed from February 1992 through January 1994 were identified through the population-based Shanghai Cancer Registry. A control group of 601 nonsmoking women was selected randomly from the Shanghai Residential Registry, and they were frequency-matched to the expected age distribution of the cases. Exposure to indoor air pollutants from Chinese-style cooking was ascertained through in-person interviews. We estimated adjusted odds ratios (OR) and 95% confidence intervals (CI) by unconditional logistic regression. There were similar patterns of excess risk for exposure to indoor air pollutants from Chinese-style cooking across different histological types of lung cancer. Women who did not have a separate kitchen experienced a 28% increased risk of lung cancer (OR = 1.28; 95% CI = 0.98-1.68). We found little association with area of the windows of the apartment where subjects had lived for the longest period of time. Heating cooking oils to high temperatures was associated with a 1.64-fold increased risk of lung cancer (95% CI = 1.24-2.17). An 84% excess risk was found among women who most often cooked with rapeseed oil (OR = 1.84; 95% CI = 1.12-3.02). Lung cancer risks were also related to "considerable" smokiness of the kitchen during cooking (OR = 2.38; 95% CI = 1.58-3.57), frequent eye irritation during cooking (OR = 1.68; 95% CI = 1.02-2.78), to a more than weekly use of frying (OR = 2.09; 95% CI = 1.14-3.84) and deep-frying (OR = 1.88; 95% CI = 1.06-3.32). This population-based case-control study confirmed that exposure to indoor air pollution from Chinese-style cooking, especially cooking unrefined rapeseed oil at high temperatures in woks, may increase the risk of lung cancer. PMID- 10468421 TI - Health deficiencies in Cape Breton County, Nova Scotia, Canada, 1950-1995. AB - Cape Breton County contains one of the most polluted areas in North America and is socioeconomically depressed. We evaluated mortality patterns in this area over the past 5 decades, focusing on life expectancy and life loss. Life loss refers to the difference in life expectancy of Cape Breton County residents and all Canadians, and was further broken down into disease-specific components using cause-eliminated life table methods. We observed lags in health of 20 to 25 years for residents of Cape Breton County. Life expectancy in some municipalities of Cape Breton County is reduced by more than 5 years. Life loss for these residents is greater than that of any single cause of death for Canadians. Life loss among Cape Breton County women is primarily attributable to cancer, and, among men, to cardiovascular diseases. Life loss from cancer is higher in the steel-producing communities; whereas life loss from respiratory diseases and lung cancer is higher in the coal mining communities. These (and other) decompositions of life loss disclose patterns in health deficiencies that give rise to etiologic hypotheses and provide clues and directions for prevention and interventions. PMID- 10468422 TI - Smoking cessation and mortality trends among 118,000 Californians, 1960-1997. AB - We assessed the impact of smoking cessation on subsequent death rates among a cohort of 51,343 men and 66,751 women in California enrolled in late 1959 in the original American Cancer Society (ACS) Cancer Prevention Study (CPS I) and followed for 38 years. We compared the age-adjusted death rate, expressed as deaths per 1,000 person-years, among all subjects who smoked cigarettes in 1959 but who had largely quit as of 1997 with the death rate among never smokers over a 38-year period. The all causes death rate for males decreased from 20.67 during 1960-1969 to 18.68 during 1960-1997 for smokers and decreased from 10.51 to 9.46 for never smokers. The lung cancer death rate for males increased from 1.558 to 1.728 for smokers and increased from 0.127 to 0.133 for never smokers. The all causes death rate for females increased from 9.54 to 10.14 for smokers and decreased from 6.95 to 6.44 for never smokers. The lung cancer death rate for females increased greatly from 0.208 to 0.806 for smokers and increased from 0.094 to 0.116 for never smokers. These results indicate there has been no important decline in either the absolute or relative death rates from all causes and lung cancer for cigarette smokers as a whole compared with never smokers in this large cohort, in spite of a substantial degree of smoking cessation. While cessation clearly reduces the mortality risk among long-term former smokers, the population impact of cessation appears to be less than currently believed. PMID- 10468423 TI - Water chlorination and birth defects. AB - Chlorination of drinking water that contains organic compounds leads to the formation of by-products, some of which have been shown to have mutagenic or carcinogenic effects. As yet, too little is known about the possible teratogenic effects on the human fetus. We linked the Norwegian waterwork registry, containing 1994 data on chlorination practice and color (an indicator for natural organic matter), with the Medical Birth Registry for 1993-1995. The proportion of the population exposed to chlorination and a weighted mean color number in drinking water was computed for each municipality. Among 141,077 births, 2,608 (1.8%) had birth defects. In a comparison between exposed (high color; chlorination) and reference groups (low color; no chlorination), the adjusted odds ratio was 1.14 (0.99-1.31) for any malformation, 1.26 (0.61-2.62) for neural tube defects, and 1.99 (1.10-3.57) for urinary tract defects. This study provides further evidence of the role of chlorination of humic water as a potential cause of birth defects, in a country with relatively low levels of chlorination byproducts. PMID- 10468424 TI - Familial occurrence of preeclampsia. AB - We conducted a cohort study on whether preeclampsia during the pregnancy of a mother is a risk factor for preeclampsia during the pregnancy of her daughter. Data from the Medical Birth Registry were combined with data from a local registry of births from 1955 to 1990. We could identify the births of 22,768 elder daughters and 2,959 younger daughters. These daughters had also experienced at least one delivery. If the mother had preeclampsia during her pregnancy with an elder daughter, then the elder daughter had an increased risk for preeclampsia in her first pregnancy (relative risk (RR) = 1.7; 95% confidence interval (CI) = 1.3-2.2). This increased risk persisted into the elder daughter's second pregnancy (RR = 1.7; 95% CI = 1.1-2.6). The risks for the daughters were also increased to a similar level if the mother had experienced preeclampsia in any other pregnancy. Furthermore, the risks were similarly elevated if only mothers with firstborn children were included in the analyses. Hence, preeclampsia during the pregnancy of a mother was a risk factor for development of preeclampsia during the pregnancy of her daughters; however, owing to a relatively small population attributable proportion, genetic predisposition explained only a minor part of the occurrence of preeclampsia in this population. PMID- 10468425 TI - Adult glioma in relation to residential power frequency electromagnetic field exposures in the San Francisco Bay area. AB - In a population-based study, we examined residential power frequency electromagnetic field exposures for 492 adults newly diagnosed with histologically confirmed glioma between August 1, 1991 and April 30, 1994, in the San Francisco Bay area and 462 controls, obtained through random-digit dialing frequency, matched to cases for age, gender, and race. Residential exposure assessment consisted of spot measures with EMDEX (Enertech Consultants, Campbell, CA) meters and wire codes based on characterization and location of nearby power lines. We considered the index residence at the time of the case's diagnosis or the control's interview and all other California residences of each subject for 7 years before study entry. We obtained wire codes for eligible residences of 76% and for index residences of 99% of subjects. Using the Kaune-Savitz wire code classification, the relative risk for longest held residences coded as "high" compared with "low" was 0.9 [95% confidence interval (CI) = 0.7-1.3], while relative risk and 95% CIs for front door spot measures of 1.01-2 milligauss, 2.01 3 milligauss, and higher than 3 milligauss compared with < or =1 milligauss were 1.0 (0.7-1.4), 0.6 (0.3-1.1), and 1.7 (0.8-3.6). Adjustment for age, gender, race, and whether the subject owned the residence did not meaningfully alter these findings, nor did comparisons using index or highest coded residence. Because of potential exposure misclassification and the unknown pertinent exposure period, these data cannot provide strong support against, but clearly do not support an association between, adult glioma and residential power frequency electromagnetic field exposures. PMID- 10468426 TI - Menstrual and reproductive factors for salivary gland cancer risk in women. AB - Several observations suggest that salivary gland cancer may be, in part, a hormonally dependent disease. We examined associations between hormonally mediated life events and salivary gland cancer risk in a population-based case control study. Of 76 women diagnosed between 1989 and 1993, 63 (83%) were interviewed. Of 111 population controls, 83 (75%) were interviewed. Early menarche (OR = 4.1, 95% CI = 1.4-12.1) and nulliparity (OR = 2.6, 95% CI = 1.0 6.7) were associated with increased risk whereas late age at first full-term pregnancy (OR = 0.19, 95% CI = 0.3-1.2) and longer duration of oral contraceptive use (OR = 0.31, 95% CI = 0.10-1.0) were associated with diminished risk. These findings are consistent with a hormonal component in salivary gland cancer risk. PMID- 10468427 TI - Radiation exposure and cancer mortality in uranium processing workers. AB - Data from the Comprehensive Epidemiology Data Resource (CEDR) allowed me to study patterns of cancer mortality in a cohort of 4,014 uranium-processing workers. Employing risk-set analysis for cohort data, I estimated the effects of external (gamma) and internal (alpha) radiation on cancer mortality. My results indicate that Fernald workers exposed to ionizing radiation experienced an increase in mortality from total cancer (per 100 mSv external dose rate ratio (RR) = 1.92; 95% confidence interval (CI) = 1.11-3.32), radiosensitive solid cancer (RR = 2.00; 95% CI = 1.02-3.94), and lung cancer (RR = 2.77; 95% CI = 1.29-5.95). Effects were strongest when exposure had occurred at older ages (>40 years). In addition, I observed an increase in lung-cancer mortality for workers exposed to > or =200 mSv of internal (alpha) radiation (RR = 1.92; 95% CI = 0.53-6.96). Furthermore, my results demonstrate the importance of a long follow-up time when studying solid cancers, the potential for bias due to worker selection associated with concomitant chemical exposures, problems of exposure measurement, confounding, and effect modification due to age at exposure. Owing to lack of data, a previous pooled analysis of uranium-processing workers could only partially address these issues. PMID- 10468428 TI - Control for seasonal variation and time trend in case-crossover studies of acute effects of environmental exposures. AB - The case-crossover study design is used to study the triggers of acute outcomes in populations. It controls for all measured and unmeasured time-invariant confounders by design. Studies of environmental triggers of morbidity are potentially confounded by temporal trends in the outcome owing to omitted covariates. We conducted a simulation study of the case-crossover design's ability to control for temporal confounding patterns by design rather than through modeling. We compared five case-crossover control sampling strategies including the matched pair, a symmetric bi-directional, a total history approach, and two approaches proposed by Navidi (Biometrics 1998;54:596-605). We simulated true relative risks (RR) of 1.10 and 2.00 and induced confounding by seasonal patterns as well as linear and nonlinear long-term trends to yield estimated RR values as high as 3.18. The symmetric bi-directional approach was compared across four lag times and controlled for temporal confounding best when the lag was shortest. With a 1-week lag, it estimated the RR values as 1.10 and 2.01. The four other approaches failed to control for the temporal trends. Our simulations show that the symmetric bi-directional case-crossover design can substantially control for temporal confounding by design although it is not as efficient (66%) as Poisson regression analysis. PMID- 10468429 TI - Exposure-dependent misclassification of exposure in interaction analyses. AB - The objectives of this paper are to analyze the consequences of exposure misclassification on effect estimates in interaction analysis, and to develop a mathematical equation for the potentially biased estimate. The main point is to identify situations in which misclassification of the first exposure, dependent on the second exposure but independent on outcome status, leads to overestimation or underestimation of the interaction effect. We show that misclassification theoretically can cause overestimation of the interaction effect. Nevertheless, because the categories that yield overestimation due to misclassification are fewer than the categories that yield underestimation, and misclassification in reality mostly is multidimensional (more than one category are biased simultaneously), it is more likely that the effect of misclassification is underestimation rather than overestimation. Misclassification in the categories that lead to overestimation is compensated by misclassification in the categories that lead to underestimation. The magnitude of the biased estimate depends on the prevalences of the misclassified exposure, stratified for the second exposure and the outcome-the lower the prevalence, the smaller the bias. PMID- 10468431 TI - Long-term follow-up of cognitive outcome after breech presentation at birth. AB - Studies of long-term consequences of birth in breech presentation are sparse. Therefore, we conducted a cohort study linking birth registry data with data collected during evaluation for military service in 4,298 conscripts born between 1973 and 1976. The cognitive functions were measured with the Boerge Prien IQ test. A total of 164 conscripts were born in breech presentation and 70 (42.7%) of these were delivered after Caesarean section. The mean Boerge Prien test score was 43.2 among men born in cephalic presentation and 39.9 among those born in breech presentation for a difference of 3.3 (95% confidence interval = 1.8-4.7). The negative association between breech presentation and cognitive outcome persisted after stratifying by Caesarean section and after adjustment for confounders. It also persisted when we restricted the analyses to term singleton pregnancies. PMID- 10468430 TI - Fine particles and peak flow in children: acidity versus mass. AB - We assessed the impact of summertime haze episodes on twice daily peak flow measurements of children in Philadelphia, Pennsylvania. Height-adjusted peak flows were regressed on weather and air pollution concentrations. Lower morning peak flows were associated with exposure to inhalable particulate matter (-2.94 liters/minute/18 microg/m3, 95% confidence limits (CL) = -0.56, -5.33), and fine sulfate particles (-2.44 liters/minute/8 microg/m3, 95% CL = -0.36, -4.51). Particle-strong acidity and the coarse particle mass were weakly associated with lower peak flow. Acutely lower peak flows in children were associated with fine sulfate particles, but only weakly with the acidity of the fine particles. PMID- 10468432 TI - Nuclear weapons testing and research efforts to evaluate health effects on exposed populations in the United States. PMID- 10468433 TI - Capture-recapture estimation using statistical software. PMID- 10468434 TI - Parental smoking and Helicobacter pylori in children. PMID- 10468435 TI - Estimation of relative and absolute test accuracy. PMID- 10468436 TI - Improved decision-making in environmental and occupational health. PMID- 10468437 TI - How much global ill health is attributable to environmental factors? AB - Over the years, estimates have been made of the portions of human mortality and morbidity that can be attributed to environmental factors. Frustratingly, however, even for a single category of disease such as cancer, these estimates have often varied widely. Here we attempt to explain why such efforts have come to such different results in the past and to provide guidance for doing such estimates more consistently in the future to avoid the most important pitfalls. We do so by carefully defining what we mean by the terms "environmental," "ill health," and "attributable." Finally, based on these recommendations, we attempt our own estimate, appropriately qualified according to the many remaining uncertainties. Our estimate is that 25-33% of the global burden of disease can be attributed to environmental risk factors. Children under 5 years of age seem to bear the largest environmental burden, and the portion of disease due to environmental risks seems to decrease with economic development. A summary of these estimates first appeared in the 1997 report, "Health and Environment in Sustainable Development," which was the World Health Organization's contribution to the 5-year anniversary of the Rio Earth Summit. A full explanation of how these estimates were made is first presented here. We end with a call for a program of "strategic epidemiology," which would be designed to fill important gaps in the understanding of major environmental health risks in important population groups worldwide. PMID- 10468438 TI - Methodologic issues in epidemiologic risk assessment. AB - This paper reviews methodologic issues pertinent to the application of epidemiology in risk assessment and discusses concerns in the presentation of results from such an activity. Assessment of the health risks associated with occupational and environmental exposures involves four phases: hazard identification, i.e., the detection of the potential for agents to cause adverse health effects in exposed populations; exposure assessment, i.e., the quantification of exposures and the estimation of the characteristics and sizes of the exposed populations; dose-response assessment, i.e., the modeling for risk realization; and risk characterization, i.e., the evaluation of the impact of a change in exposure levels on public health effects. The risk-assessment process involves limitations of exposure data, many assumptions, and subjective choices that need to be considered when using this approach to provide guidance for health policy or action. In view of these uncertainties, we suggest that the provision of estimates of individual risk and disease burden in a population must be accompanied by the corresponding estimates of precision; risks should be presented in a sufficiently disaggregated form so that population heterogeneities are not lost in the data aggregation; and different scenarios and risk models should be applied. The methods are illustrated by an assessment on the health impacts of exposure to silica. PMID- 10468439 TI - On the comparable quantification of health risks: lessons from the Global Burden of Disease Study. AB - Extensive discussion and comments on the Global Burden of Disease Study findings have suggested the need to examine more carefully the basis for comparing the magnitude of different health risks. Attributable burden can be defined as the difference between burden currently observed and burden that would have been observed under an alternative population distribution of exposure. Population distributions of exposure may be defined over many different levels and intensities of exposure (such as systolic or diastolic blood pressure on a continuous scale), and the comparison distribution of exposure need not be zero. Avoidable burden is defined as the reduction in the future burden of disease if the current levels of exposure to a risk factor were reduced to those specified by the counterfactual distribution of exposure. Choosing the alternative population distribution for a variable, the counterfactual distribution of exposure, is the critical step in developing a more general and standardized concept of comparable, attributable, or avoidable burden. We have identified four types of distributions of exposure that could be used as the counterfactual distributions: theoretical minimum risk, plausible minimum risk, feasible minimum risk, and cost-effective minimum risk. Using tobacco and alcohol as examples, we explore the implications of using these different types of counterfactual distributions to define attributable and avoidable burden. The ten risk factor assessments included in the Global Burden of Disease Study reflect a range of methods and counterfactual distributions. We recommend that future assessments should focus on avoidable and attributable burden based on the plausible minimum risk counterfactual distribution of exposure. PMID- 10468440 TI - An aggregate public health indicator to represent the impact of multiple environmental exposures. AB - We present a framework to aggregate divergent health impacts associated with different types of environmental exposures, such as air pollution, residential noise, and large technologic risks. From the policy maker's point of view, there are at least three good reasons for this type of aggregation: comparative risk evaluation (for example, setting priorities), evaluation of the efficiency of environmental policies in terms of health gain, and characterizing health risk associated with geographical accumulation of multiple environmental exposures. The proposed impact measure integrates three important dimensions of public health: life expectancy, quality of life, and number of people affected. Time is the unit of measurement. "Healthy life years" are either lost by premature death or by loss of quality of life, measured as discounted life years within a population. Severity weights (0 for perfect health, 1 for death) are assigned to discount the time spent with conditions associated with environmental exposures. We combined information on population exposure distribution, exposure response relations, incidence, and prevalence rates to estimate annual numbers of people affected and the duration of the condition, including premature death. Using data from the fourth Dutch National Environmental Outlook, we estimated that the long term effects of particulate air pollution account for almost 60% of the total environment-related health loss in the Netherlands as modeled here. Environmental noise accounts for 24%, indoor air pollution (environmental tobacco smoke, radon, and dampness, as well as lead in drinking water) for around 6%, and food poisoning (or infection) for more than 3%. The contribution of this set of environmental exposures to the total annual burden of disease in the Netherlands is less than 5%. PMID- 10468441 TI - Environmental health impact assessment: evaluation of a ten-step model. AB - "Environmental impact assessment" denotes the attempt to predict and assess the impact of development projects on the environment. A component dealing specifically with human health is often called an "environmental health impact assessment." It is widely held that such impact assessment offers unique opportunities for the protection and promotion of human health. The following components were identified as key elements of an integrated environmental health impact assessment model: project analysis, analysis of status quo (including regional analysis, population analysis, and background situation), prediction of impact (including prognosis of future pollution and prognosis of health impact), assessment of impact, recommendations, communication of results, and evaluation of the overall procedure. The concept was applied to a project of extending a waste disposal facility and to a city bypass highway project. Currently, the coverage of human health aspects in environmental impact assessment still tends to be incomplete, and public health departments often do not participate. Environmental health impact assessment as a tool for health protection and promotion is underutilized. It would be useful to achieve consensus on a comprehensive generic concept. An international initiative to improve the situation seems worth some consideration. PMID- 10468442 TI - Global burden of disease and injury due to occupational factors. AB - We made estimates of absolute morbidity and mortality due to occupational factors for the world using all available published data as of 1994, and, where no data were available, applying the most appropriate (in terms of similar economy, race, and environment) age-/sex-/diagnosis-specific incidence and mortality rates to known working population distributions. We report results according to economic groupings determined by the World Bank (World Development Report, 1993) and disease and injury groupings according to The Global Burden of Disease project (1997). This was part of a larger study that estimated the total global disease burden. We present aggregate results and analyses by region and disease. We estimate that approximately 100,000,000 occupational injuries (100,000 deaths) and 11,000,000 occupational diseases (700,000 deaths) occur in the world each year. We regard these as very conservative estimates which, although unavoidably crude, can nevertheless provide a basis for health priority planning at global level. PMID- 10468443 TI - Assessment of the health impact of occupational risk in Africa: current situation and methodological issues. AB - This paper presents information from the current monitoring systems in Africa, mainly Southern Africa, for occupational illness and injury and discusses the quality of the reported data in estimating the health impact of occupational risk. The paper presents and discusses the current profile of reported injury and fatalities for those countries for which data are available, in particular for the countries of the Southern African Development Community. These data indicate that the reported annual injury rates for wage workers in the Southern African Development Community region range widely from 0.35 to 49.42 injuries per 1,000 workers, and reported occupational fatality in the region ranges from 0.85 to 21.6 fatalities per 100,000 workers. Despite wide variability in reported rates (probably caused by variability in coverage and accuracy of reporting systems), transport, agriculture, mining and, to a lesser extent, construction consistently make up about three-quarters of all fatalities, with vehicle- or transport related causes accounting for high proportions of fatal accidents. The paper identifies and discusses major sources and direction of bias and error in the reported data and suggests approaches for a better assessment of the health impact of occupational illness, injury, and mortality in African countries. PMID- 10468444 TI - Global estimates of fatal occupational accidents. AB - Data on occupational accidents are not available from all countries in the world. Furthermore, underreporting, limited coverage by reporting and compensation schemes, and non-harmonized accident recording and notification systems undermine efforts to obtain worldwide information on occupational accidents. This paper presents a method and new estimated global figures of fatal accidents at work by region. The fatal occupational accident rates reported to the International Labour Office are extended to the total employed workforce in countries and regions. For areas not covered by the reported information, rates from other countries that have similar or comparable conditions are applied. In 1994, an average estimated fatal occupational accident rate in the whole world was 14.0 per 100,000 workers, and the total estimated number of fatal occupational accidents was 335,000. The rates are different for individual countries and regions and for separate branches of economic activity. In conclusion, fatal occupational accident figures are higher than previously estimated. The new estimates can be gradually improved by obtaining and adding data from countries where information is not yet available. Sectoral estimates for at least key economic branches in individual countries would further increase the accuracy. PMID- 10468445 TI - Public health and the Air Management Information System (AMIS). AB - This paper discusses the importance of public health studies with respect to risk assessment and risk management in the framework of air quality management. This is performed with respect to the Air Management Information System (AMIS), which was set up recently by the World Health Organization. The Air Management Information System is an information-exchange system in the scheme of the Global Air Quality Partnership providing information on all issues of air quality management between its participants: municipalities, countries' environmental protection agencies, international organizations, World Bank and international development banks, and nongovernmental organizations. Public health studies of air pollution-induced health effects are an important ingredient for decisions with respect to the management of air quality. First, they are to be used to derive air quality standards from air quality guidelines. Secondly, they serve to assess the causal link between observed health effects in the population and the causative agents in the air. Thirdly, they can be used to estimate ideal (in the sense of not being expressed in monetary terms) or economic damage functions that are necessary to assess the magnitude of the ideal or economic damages to human health. The latter are necessary for a sensible cost-benefit analysis in which the costs of control measures to reduce air pollution are compared with the costs of health effects. PMID- 10468446 TI - Health, environment and sustainable development: identifying links and indicators to promote action. AB - This paper discusses the links among health, environment, and sustainable development and presents a framework that extends from the epidemiological domain to the policy domain and includes the driving forces that generate environmental pressures, creating changes in the state of the environment and eventually contributing to human exposures. Health effects are the end result of this complex net of events. Environmental health interventions should not be limited to treatment of cases and directly reducing human exposures. The paper discusses the need for integrated action at all levels and, in particular, on the need to focus on long-term action directed at reducing the driving forces that generate the environmental health threats. Only this approach can achieve sustained health benefits and environmental protection in accord with the principles of sustainable development. PMID- 10468447 TI - Osseointegration? PMID- 10468448 TI - Reduction of HIV-associated oral lesions after highly active antiretroviral therapy. PMID- 10468449 TI - TMD symptoms in healthy Greek subjects. PMID- 10468450 TI - Polypoid mass of the gingiva. PMID- 10468451 TI - Oral cancer: complications of therapy. PMID- 10468452 TI - A technique to improve predictability of condylar position with modified condylotomy. PMID- 10468453 TI - Association between arthroscopic diagnosis of temporomandibular joint osteoarthritis and synovial fluid nitric oxide levels. AB - OBJECTIVE: The purpose of this study was to determine whether there is a relationship between synovial fluid levels of nitric oxide and clinical and arthroscopic findings of synovitis or cartilaginous degeneration. STUDY DESIGN: Arthroscopic surgery was performed on 20 joints in 15 female patients with internal derangement and osteoarthritis of the temporomandibular joint. Synovial fluid aspirates were obtained immediately before arthroscopy. Synovial fluid was also obtained from 14 joints of 11 female asymptomatic volunteers. The concentration of nitrite in the fluid recovered from each temporomandibular joint was measured through use of a highly sensitive and specific chemiluminescence detection method, calibrated per 1 mg of synovial fluid protein and expressed as nitric oxide; the result was then compared with clinical and arthroscopic findings of synovitis and cartilaginous degeneration. RESULTS: Significantly higher levels of nitric oxide (median, 0.331 micromol/mg) were seen in the patients with internal derangement and osteoarthritis than in the control group (median, 0.001 micromol/mg; P<.0001). Synovial fluid from joints with pain in the joint area had significantly higher levels of nitric oxide than did fluid from joints without such pain. Synovial fluid from joints with degenerative changes (median, 0.467 micromol/mg) had significantly higher levels of nitric oxide than did fluid from joints without osteoarthritis (median, 0.057 micromol/mg; P<.05). Although the levels of nitric oxide in synovial fluid aspirates were markedly elevated in some joints with synovitis, there was no correlation between the levels of nitric oxide and the presence of synovitis. CONCLUSIONS: The findings indicate that increased levels of nitric oxide are involved in the pathogenesis of cartilaginous degeneration of the temporomandibular joint. PMID- 10468454 TI - Dental extractions in patients maintained on continued oral anticoagulant: comparison of local hemostatic modalities. AB - OBJECTIVE: The purpose of this study was to evaluate postoperative bleeding in patients treated with oral anticoagulant drugs who underwent dental extractions without interruption of the treatment and to compare the effect of 3 different hemostatic modalities. STUDY DESIGN: A total of 150 patients who underwent dental extractions were divided into 3 groups. Local hemostasis was carried out as follows: group 1 (119 extractions), with gelatin sponge and sutures; group 2 (117 extractions), with gelatin sponge, sutures, and mouthwash with tranexamic acid; group 3 (123 extractions), with fibrin glue, gelatin sponge, and sutures. RESULTS: Of 150 patients, 13 (8.6%) presented with postoperative bleeding: 3 patients from group 1, 6 patients from group 2, and 4 patients from group 3. CONCLUSIONS: Dental extractions can be performed without interruption in patients treated with oral anticoagulant. Local hemostasis with gelatin sponge and sutures is sufficient. PMID- 10468455 TI - Relationship of tumor thickness in punch biopsy and subsequent surgical specimens in stage I squamous cell carcinoma of the lower lip. AB - OBJECTIVE: Among the histologic features of squamous cell carcinoma of the lower lip, maximum tumor thickness in particular is a predictor of regional nodal metastatic spread and thus an important parameter in treatment planning. The purpose of this study was to investigate the relationship between maximum tumor thickness in punch biopsies and maximum tumor thickness in subsequent surgical specimens. STUDY DESIGN: This retrospective study examined the relationship between maximum tumor thickness in punch biopsies with that in subsequent surgical specimens obtained in 72 patients with clinical stage I squamous cell carcinoma of the lower lip. RESULTS: A correlation between maximum tumor thickness in punch biopies and in subsequent surgical specimens was found only for tumors with a thickness less than 3 mm. CONCLUSIONS: Reliable predictive information could be obtained from punch biopsies with a maximum tumor thickness less than 3 mm. When the maximum tumor thickness exceeds 3 mm, better information may be obtained from either a large incisional biopsy or the surgical specimen. PMID- 10468456 TI - The effects of operator technique and bur design on temperature during osseous preparation for osteosynthesis self-tapping screws. AB - OBJECTIVE: Although the importance of atraumatic preparation of bone before the placement of osseointegrated implants has long been emphasized, the situation during placement of self-tapping screws has received little attention. Because the production of excessively high temperatures during osseous drilling is known to impair bony regeneration, the strict thermal criteria that are set for implant placement should theoretically be extended to trauma and orthognathic internal fixation. STUDY DESIGN: Temperatures achieved during the drilling of thawed human cadaveric bone before the insertion of osteosynthesis self-tapping screws were measured in vivo with thermocouples. Combinations of 2 drilling systems and 2 bur designs were studied. Variations in surgical technique were incorporated by altering both the force applied by the operator and the application of irrigant. Statistical significance was examined through use of a Student t test. RESULTS: One of the 2 bur designs and careful operator technique were associated with significantly smaller increases in temperature. Irrigation had the greatest effect on temperature recorded, the absence of irrigant resulting in temperatures in excess of 70 degrees C. CONCLUSIONS: These findings emphasize the importance of careful surgical technique and constant irrigation during osseous preparation if the potential for bone regeneration around osteosynthesis self-tapping screws is to be maximized. PMID- 10468457 TI - The role of the disk in sheep temporomandibular joint ankylosis. AB - OBJECTIVE: The purpose of this study was to determine the role of the disk in intraarticular ankylosis of the temporomandibular joint. STUDY DESIGN: Twelve adult sheep were divided into 2 groups. In group 1, removal of the temporal and condylar articular surfaces was performed on the right temporomandibular joint and the disk was maintained; in group 2, removal of the articular surfaces and diskectomy were performed on the right temporomandibular joint. One sheep from each group was killed just after surgery and 5 sheep from each group were killed at 3 months. The joints were examined radiologically, macroscopically, and histologically. The range of jaw movements was recorded preoperatively and at sacrifice. RESULTS: Each of 2 sheep in group 2 had lost 4% of their body weight by 3 months; all of the other sheep maintained or increased their weight. The range of jaw motion to the right was significantly lower in group 2 than in group 1 (P<.01). In group 1, fibrous repair of the articular surface and regeneration of the condylar head was seen. In group 2, each of the joints showed a total fibrous ankylosis with some calcification. There was a statistically significant difference in radiologic score between the groups (P<.0001). Histologic scores for group 1 demonstrated significantly lower scores on the degree of ankylosis scale and degree of calcification scale (P<.0001). CONCLUSIONS: This study showed that the presence of the disk prevented the development of fibrous intraarticular ankylosis of the temporomandibular joint. PMID- 10468458 TI - A retrospective study of the management of oral mucous membrane pemphigoid with dapsone. AB - OBJECTIVE: The purpose of this retrospective study was to investigate the efficacy of dapsone therapy in the management of mucous membrane pemphigoid. STUDY DESIGN: The charts of 29 patients who had been diagnosed with mucous membrane pemphigoid by means of routine histologic analysis and direct immunofluorescence were reviewed. The oral features were graded according to severity of disease from 1 to 3. Each patient was assigned to one of 4 groups according to his or her response to therapy. RESULTS: Nine patients were treated successfully with topical corticosteroids alone. Eleven patients with moderate to severe disease who were treated initially with topical steroids showed minimal improvement; after dapsone was added, 7 of the 11 had total resolution of their lesions and 4 had greater than 75% improvement. Two patients had to discontinue dapsone because of side effects. CONCLUSIONS: In this group of 20 patients with moderate to severe mucous membrane pemphigoid, the use of dapsone in combination with topical corticosteroids caused greater than 75% resolution of oral lesions in all patients studied. PMID- 10468459 TI - A dentoalveolar abscess in a pediatric patient with ketoacidosis caused by occult diabetes mellitus: a case report. AB - Oral health professionals are frequently asked to evaluate patients with routine odontogenic infections. These patients can sometimes present with systemic signs and symptoms, including fever, malaise, tachycardia, and dehydration. It is important for the astute clinician to understand the possible associated systemic diseases that may be contributing to odontogenic infections. We present here an interesting case of a pediatric patient with a routine canine space infection who exhibited classic clinical signs and symptoms of diabetic ketoacidosis. PMID- 10468460 TI - Lip adhesion: an unusual complication of erythema multiforme. AB - The purpose of this article is to report a case of lip adhesion as a complication of erythema multiforme in an 8-year-old child. This is the second reported case of this complication of oral ulceration associated with erythema multiforme. PMID- 10468461 TI - Pulmonary metastasis of ameloblastoma: case report and review of the literature. AB - Ameloblastomas are benign tumors of odontogenic epithelial origin. There is a high incidence of local recurrence associated with these tumors, and distant metastasis is rare. A review of the English literature shows that there have been 41 prior reports of pulmonary metastases from ameloblastomas of the oral cavity. We present another case of ameloblastoma metastatic to the lung and review the histopathology and mechanism of metastatic spread. PMID- 10468462 TI - Dedifferentiation occurring in adenoid cystic carcinoma of the tongue. AB - A 61-year-old man came to the University of Granada School of Dentistry complaining of a mass involving his ventral tongue. Histopathologic examination of the excised specimen showed adenoid cystic carcinoma in which cribriform and tubular patterns were observed, juxtaposed with an undifferentiated carcinoma, large-cell type. No cervical lymph node metastasis was present, and the patient is alive and free of disease 5 years after treatment. To our knowledge, no similar cases have been reported thus far, though other salivary gland malignancies have been described in association with undifferentiated carcinoma, especially in the parotid gland. These neoplasms have been highly aggressive, and the adequacy of the primary surgical resection may be critical in determining the ultimate prognosis and survival. PMID- 10468463 TI - Actinic cheilitis: a review of 152 cases. AB - OBJECTIVE: The purpose of this study was to determine whether any clinical or histopathologic variables are associated with the severity of epithelial change in lesions of actinic cheilitis. STUDY DESIGN: A total of 152 acceptable cases of actinic cheilitis were identified from 66,067 cases accessioned from February 1989 to June 1998. For each case, the clinical information supplied by the submitting practitioner at the time of the biopsy and 8 histopathologic variables were evaluated. RESULTS: The following 5 histopathologic variables were positively correlated with an increased degree of epithelial change: acanthosis, basophilic change within the connective tissue, the presence of inflammation within the connective tissue, perivascular inflammation, and thickness of the keratin layer. None of the clinical variables was associated with an increased degree of epithelial change. CONCLUSIONS: The presence of any of the aforementioned histopathologic changes should prompt a close evaluation of the lesion for the presence of either epithelial dysplasia or carcinoma. PMID- 10468464 TI - Quantitative assessment of apoptosis in oral lichen planus. AB - OBJECTIVE: The aims of this study were to examine the frequency of apoptoses in oral lichen planus by in situ end labeling, to ascertain whether this technique is as sensitive as conventional histologic analysis, and to examine the effect of lymphocytic infiltration. STUDY DESIGN: Numbers of apoptoses in hematoxylin-eosin stained sections were compared with numbers of apoptotic nuclei identified by in situ end labeling in oral lichen planus (n = 26) and normal buccal epithelium (n = 8). Immunohistochemical staining with MIB-1 and for Bcl-2 and Bax enabled possible regulatory pathways to be investigated. RESULTS: In oral lichen planus, approximately 1 apoptotic cell was detected per millimeter of basal layer, cell death increasing with lymphocytic infiltration. Epithelial cell proliferation did not correlate with apoptosis. Bcl-2 expression was weak or absent in basal cells, and Bax was localized to upper prickle cells. CONCLUSIONS: Increased numbers of apoptoses were detected in oral lichen planus, especially in association with lymphocytic infiltration, higher numbers being seen with hematoxylin-eosin staining than with in situ end labeling. PMID- 10468465 TI - Linear IgA disease histopathologically and clinically masquerading as lichen planus. AB - In each of 2 cases reported, the patient presented with features of erosive lichen planus or lichenoid drug eruptions and an incisional biopsy taken from the patient was diagnosed histologically as lichen planus. Subsequent recurrences or exacerbations were associated with vesiculobullous lesions. Simultaneous or subsequent direct immunofluorescence studies--from the same tissue sample in one case and from a similar site in the other case--demonstrated classic features of linear IgA disease. Both patients were originally treated for lichen planus with systemic and/or topical corticosteroids with limited success. One patient was treated with sulfapyridine with minimal improvement. Both patients were subsequently treated with dapsone and demonstrated significant clinical improvement. We propose that linear IgA disease may be more common than reported in the oral cavity, inasmuch as many cases of recalcitrant lichen planus, erosive lichen planus, and lichenoid drug eruptions, especially those with a vesiculobullous component, may in reality represent linear IgA disease. We recommend that direct immunofluorescence be done in any case in which bullous lichen planus is suspected. PMID- 10468466 TI - Increased interleukin-8 expression in inflamed human dental pulps. AB - OBJECTIVE: Elevated levels of interleukin-8, a potent chemoattractant and activator of neutrophils, are associated with infectious and inflammatory diseases. However, little is known about interleukin-8 expression in human dental pulp. The purpose of this study was to determine whether tissue levels of interleukin-8 are elevated in irreversibly inflamed human pulps. STUDY DESIGN: Experimental samples were from teeth clinically diagnosed with irreversible pulpitis (diseased pulps). Controls were from freshly extracted, caries-free third molars (normal pulps). Samples were subjected to enzyme-linked immunosorbent assay and/or immunohistochemical analysis with specific antibodies to interleukin-8. RESULTS: The enzyme-linked immunosorbent assay studies showed elevated levels of interleukin-8 in diseased pulps (mean, 1.82+/-0.79 pg/mL/microg protein), as compared to detectable interleukin-8 levels in samples from normal pulps (mean, 0.08+/-0.04 pg/mL/microg protein; P<.05). Immunohistochemical analyses demonstrated that diseased samples exhibited a higher density of localized interleukin-8 staining in areas with heavy infiltration of inflammatory cells. In contrast, normal pulps showed negative or weak interleukin-8 staining. CONCLUSIONS: Interleukin-8 concentration was higher in pulps diagnosed with irreversible pulpitis; only negligible amounts of interleukin-8 were present in normal pulps. PMID- 10468467 TI - Release of formaldehyde by 4 endodontic sealers. AB - OBJECTIVE: The purpose of this study was to evaluate the release of formaldehyde by some root canal filling materials. STUDY DESIGN: Two older endodontic sealers, AH 26 and Endomethasone, and 2 recently available sealers, AH Plus and Top Seal, were analyzed. Infrared and electronic spectroscopy were used to determine formaldehyde content after set of the materials. RESULTS: Analysis showed that the AH 26 and Endomethasone sealers released formaldehyde. Although the AH Plus and Top Seal sealers have similar chemical composition, they released formaldehyde in a minimal concentration. CONCLUSIONS: The AH 26 and Endomethasone sealers released formaldehyde after setting; however, a minimum release was observed for the AH Plus and Top Seal sealers. PMID- 10468468 TI - Statistical study for sonographic differential diagnosis of tumorous lesions in the parotid gland. AB - OBJECTIVE: The purpose of this study was to clarify characteristic sonomorphologic features of parotid lesions statistically and to propose new criteria for the differential diagnosis. STUDY DESIGN: Eighty-six tumorous lesions were analyzed with regard to the following sonomorphologic features: boundary, shape, echo intensity level, distribution of internal echoes, and acoustic enhancement. Stepwise polychotomous logistic regression analysis was performed to assess characteristic sonographic features. As dependent variables, we used "pleomorphic adenoma," "Warthin tumor," "malignant tumors" and "other benign lesions"; as predictor variables, we used the aforementioned sonomorphologic features. Proportion of the occurrence of each dependent variable was calculated. RESULTS: Lobular shape and homogeneous internal echoes predicted pleomorphic adenoma. A lesion with multiple anechoic areas would be Warthin tumor with very high sensitivity. Malignant tumors showed either heterogeneous internal echoes without characteristic structures or polygonal shape. CONCLUSIONS: These sonomorphologic features should be observed to make more exact differential diagnoses for operation and therapy planning. PMID- 10468469 TI - Detection of caries with conventional digital imaging and tuned aperture computed tomography using CRT monitor and laptop displays. AB - OBJECTIVE: The purpose of this study was to compare the diagnostic performance of conventional digital images and tuned aperture computed tomography (TACT) slices in caries detection through use of cathode-ray tube monitor and laptop displays. STUDY DESIGN: Forty-two extracted posterior teeth were mounted and imaged with a direct digital radiography system. Conventional digital bitewing projections and TACT slices were acquired. Images were viewed on a high-resolution cathode-ray tube monitor and on an active-matrix laptop display. Eight observers assessed caries status of occlusal and proximal surfaces of the teeth using all combinations of image and display modality. Observers' assessments were compared with the results of histologic examination of tooth sections. Possible differences in receiver operating characteristic curve areas among displays, image modalities, observers, and surfaces were analyzed by means of analysis of variance. RESULTS: There was no statistically significant difference between the diagnostic performances provided by the cathode-ray tube monitor and laptop displays in caries detection (P = .588). In addition, the performances of digital images and TACT slices were not significantly different (P = .843). CONCLUSIONS: Modern active-matrix laptop displays provide diagnostic quality for caries detection comparable to that obtainable with cathode-ray tube monitors. PMID- 10468470 TI - An in vivo comparison of diagnostic information obtained from tuned-aperture computed tomography and conventional dental radiographic imaging modalities. AB - OBJECTIVE: The purpose of this study was to compare diagnostic information obtained by means of 3-dimensional tuned-aperture computed tomography (TACT) and by means of conventional radiography of patients requiring surgery. STUDY DESIGN: TACT produced digital images that yielded a series of tomographic slices viewed interactively. Controls were conventional periapical and/or panoramic radiographs. Each of 4 independent dentists performed 2 tasks, one requiring an estimation of confidence in their clinical assessments of the patient and the other requiring an estimation of the resulting diagnostic potential for altering associated treatment options. Data were analyzed through use of the nonparametric Mann-Whitney U Wilcoxon rank sum W test. RESULTS: A statistically significant difference for both tasks was observed (2-sided; P<.001). CONCLUSIONS: TACT displays were more diagnostically informative and had more impact on potential treatment options than did conventional radiographs. PMID- 10468471 TI - Ultrasonic determination of thickness of masticatory mucosa: a methodologic study. AB - OBJECTIVE: The aim of the present study was to assess the reliability and validity of ultrasonic measurements of thickness of the masticatory mucosa. STUDY DESIGN: Eleven periodontally healthy subjects took part. Thickness of the buccal gingiva was assessed midbuccally and interdentally. In the maxilla, the palatal mucosa was measured at every tooth midpalatally, between the first and second molars, and between the second and third molars at each of 3 defined locations 4 mm apart. In the mandible, thickness of the lingual gingiva was measured midlingually as well as interdentally between the first and second molars and between the second and third molars. Thickness of the retromolar mucosa was also assessed. All 1,293 measurements were repeated after 24 hours. Validity of measurements was tested in a porcine model. RESULTS: Mean (+/- standard deviation) thickness of the midbuccal and midlingual gingiva was 0.99+/-0.52 mm; the measurement error was 0.26 mm. Measurements were well correlated (Pearson's r = .74, P<.001). Mean thickness of the papillary gingiva was 1.19+/-0.53 mm; the measurement error was 0.37 mm (r = .58, P<.001). A lower reliability was observed for palatal measurements of mucosal thickness; mean thickness was 2.36+/-0.87 mm, and measurement error was 0.54 mm (r = .64, P<.001). The validity of measurements was excellent in the 0.5-4.5 mm range. CONCLUSIONS: Differences in reliability of ultrasonic assessments of mucosal thickness in different parts of the oral cavity may depend on the difficulties of repeatedly measuring at the same location, on varying thickness of the tissues, and on expression of palatal rugae. These problems might be resolved by averaging multiple measurements. PMID- 10468472 TI - [Individualized backache. Insurance system plus over-treatment support epidemics in the 90's]. AB - Chronic back pain leading to invalidity has become a social medical problem of enormous proportions. After several months off work, efforts at rehabilitation often fail because secondary effects prevent the return to normal daily activities. For this reason, early detection and prevention of chronicity is of considerable importance. The background to the development of chronicity is complex. In addition to the wide range of somatic and psychosocial factors, the roles played by the physicians and social insurances are also examined. PMID- 10468473 TI - [Back pain--how to prevent disability. Planning for graduated therapy]. AB - Early detection and prevention of chronicity is of prime importance. In the acute phase of the condition, this involves the use of a range of simple diagnostic measures, the aim of which is to search for possible warning signs of a serious disorder with a specific cause. If the patient's complaints are unspecific, a rapid return to his/her usual daily activities should be the aim, supported by patient instruction and management of pain. When the patient's complaints persist for more than 4 weeks, a more comprehensive diagnostic work-up is indicated, with the aim of establishing the underlying cause of persisting problems. In addition to a renewed search for specific causes, this includes the identification of psychosocial factors that might be impediments to the return of the patient to his/her usual activities. For further diagnostic and therapeutic considerations, these points require particular attention. PMID- 10468474 TI - [The patient with back pain and the family physician. Problem solving with simple methods]. PMID- 10468475 TI - [CNS symptoms caused by hypoglycemia: frequent misdiagnosis: "stroke". Pitfall of the neuroglycopenia syndrome]. AB - Hypoglycemia may occur without classic symptoms, especially in elderly patients hypoglycemia may imitate nearly every neurological symptom. The neuroglycopenic syndrome, which is often due to sulfonylurea-induced hypoglycemia, is frequently overlooked or misinterpreted as cerebral ischemia. Therefore in every case of disturbance of consciousness, acute neurologic deficits and psychiatric abnormalities an immediate blood glucose test should be performed to exclude hypoglycemia. PMID- 10468476 TI - [Diagnosis and therapy of posttraumatic headache. Careful early mobilization prevents chronic condition]. PMID- 10468477 TI - [Pediculosis]. PMID- 10468478 TI - [The "Eternal President of Physicians" takes leave. Vilmar's critical last words]. PMID- 10468479 TI - [The physician's image 1999--inspite of scolding by the media. The spotless white coat]. PMID- 10468480 TI - [Transsexuality--a multidisciplinary problem. Which therapeutic methods proved to be successful?]. AB - Transsexualism as a phenomenon clearly contrasts the common understandings of sexuality. This can lead to conflicts for the transsexual himself as well as for those being confronted. Medical care and legislative offer some assistance for patients, which has been proofed as suitable and helpful. These strategies have not been developed by a theoretical approach but pragmatically and can still be further improved. To be able to receive a higher acceptance of this phenomenon in society and to achieve concrete improvement of the patient's personal situation the challenge still exists to understand mechanisms underlying the transsexual experience. PMID- 10468481 TI - [Transsexualism--when is gender transformation operation indicated?]. AB - The indications for transformation surgery for transsexuals cannot be defined at one single examination; they gradually become apparent over a period of long-term care lasting a minimum of 1.5 years, beginning with the first contact with the therapist and ending with surgery. Patients who have been well prepared for the sex change over this period usually become socially well integrated after the operation. Positive results do not depend on sexually corrective surgery alone, but on the overall therapeutic package. Although the operation represents the last step in the course of treatment for most transsexuals, surgical correction per se is not the solution for every patient with problems of sexual identity. PMID- 10468482 TI - [Sex-adjusting surgery in transsexualism]. AB - Surgery for sex reassignment is an established procedure in both female-to-male and male-to-female transsexuals. While good cosmetic and functional results can be achieved in most patients of the former group in 2 sessions, in the latter the best possible results necessitate numerous interventions and scarring is inevitable. Our own preferred surgical techniques are described in detail and illustrated. Such complex interventions should be done only in specialized centres in cooperation between urologists, gynecologists and plastic surgeons. PMID- 10468483 TI - [Advanced education by the Diabetes Society. Diabetic cheiropathy and related diseases]. PMID- 10468484 TI - [Differential diagnosis of anemia. Clinical symptoms have little diagnostic value; MMW-Seminar on anemias, 1]. PMID- 10468485 TI - [Combined hepatitis A/B immunization in children and adolescents. Economic evaluation of different immunization methods]. PMID- 10468486 TI - [Scabies]. PMID- 10468487 TI - [Diagnostic quiz: Orange-colored urine]. PMID- 10468489 TI - [Your bill will hold its ground in court]. PMID- 10468488 TI - [Health reform 2000. Hard measures against contract physicians]. PMID- 10468490 TI - [The usefulness of today's hearing aids--possibilities and limitations]. AB - Some ten to twelve million people in Germany are estimated to need hearing aids. Many of the hard of hearing suffer from inner ear deafness with hair cell dysfunction, and not only hear sounds at too low a level of intensity, but also distorted and unintelligible. Modern special digital hearing aids are able, by appropriate amplification of certain frequencies or frequency bands or, e.g. through the use of directional microphones, to improve discrimination of speech from surrounding noise. In patients with mild hearing losses in particular, the use of bilateral hearing aids is recommended to improve speech recognition. Attention is drawn to the importance of individual counselling and the testing of the hearing aids by the patient. PMID- 10468491 TI - [Hearing in old age. Hearing loss is never physiologic]. AB - The common form of hardness of hearing seen in old age is not, in the first instance, due to physiological age-related changes as is, for example, presbyopia, but rather to a complex sensorineural pattern of injury. The underlying causes are not yet fully understood. Typical medical risk factors such as hypertension, diabetes mellitus, etc. apparently play a smaller role than increasing environmental noise and modern life styles. The various forms of presbyacusis and their presentation in the pure tone audiogram are discussed. PMID- 10468492 TI - [Hearing loss made clear by the statement of a third person. Suggestions for the practice team]. PMID- 10468493 TI - [Psychosocial stress as trigger for hypertension]. PMID- 10468494 TI - [The Hypertension Ligue on criticism of its recommendations. Emphasis on Ca antagonists!]. PMID- 10468495 TI - [Diagnosis and therapy of renal anemia. Iron levels should be watched carefully: MMW Seminar on Anemia. 2]. PMID- 10468496 TI - [Single daily dose of calcium antagonist is enough. Advantage for compliance and blood pressure lowering]. PMID- 10468497 TI - [Heart failure. 1: acute heart failure]. PMID- 10468498 TI - [Diagnostic quiz. Unusual cause of ulcus vnetriculi]. PMID- 10468499 TI - [Diagnostic quiz: facial ulceration and Horner syndrome]. PMID- 10468500 TI - [Transsexualism--medical intervention can lead to correct gender]. PMID- 10468501 TI - Chronic tension-type headache: where are we? PMID- 10468502 TI - Topographical disorientation: a synthesis and taxonomy. AB - Over the last century, several dozen case reports have presented 'topographically disoriented' patients who, in some cases, appear to have selectively lost their ability to find their way within large-scale, locomotor environments. A review is offered here that has as its aim the creation of a taxonomy that accurately reflects the behavioural impairments and neuroanatomical findings of this literature. This effort is guided by an appreciation of the models of normative way-finding offered by environmental psychology and recent neuroscience research. It is proposed that several varieties of topographical disorientation exist, resulting from damage to distinct neuroanatomical areas. The particular pattern of impairments that patients evidence is argued to be consonant with the known functions of these cortical regions and with recent neuroimaging results. The conflicting claims of previous reviews of this area are also considered and addressed. PMID- 10468503 TI - Possible mechanisms of action of nitric oxide synthase inhibitors in chronic tension-type headache. AB - It has been demonstrated recently that nitric oxide synthase (NOS) inhibition has an analgesic effect in patients with chronic tension-type headache. The aim of the present study was to investigate the influence of the NOS inhibitor, L-N(G) methyl arginine hydrochloride (L-NMMA), on two of the most prominent features of chronic tension-type headache, i.e. increased muscle hardness and increased myofascial tenderness. In a double blind, crossover designed trial, 16 patients with chronic tension-type headache were randomized to receive intravenous infusion of 6 mg/kg L-NMMA or placebo on 2 days separated by at least 1 week. Muscle hardness of the trapezius muscle was measured with a hardness meter. Myofascial tenderness in the pericranial region was evaluated by manual palpation with standardized and validated methodology. All parameters were recorded at baseline and at 60 and 120 min after start of infusion. Compared with baseline, muscle hardness, 107 +/- 17 kPa/cm and tenderness, 18 +/- 11 were significantly reduced at 60 and 120 min to: hardness, 101 +/- 17 kPa/cm and 101 +/- 17 kPa/cm, respectively; tenderness, 15 +/- 11 and 14 +/- 11, respectively, after treatment with L-NMMA (P < 0.05 and P < 0.01, respectively), while there was no significant reduction at any time after treatment with the placebo. Compared with the placebo, the summary score of muscle hardness was significantly reduced (P = 0.04), while tenderness showed a non-significant reduction (P = 0.11) following treatment with L-NMMA. Since increased muscle hardness in patients with chronic tension-type headache may reflect sensitization of second order neurons due to prolonged nociceptive input from myofascial tissues, we suggest that the decrease in muscle hardness following treatment with L-NMMA may be caused by reduction of central sensitization. PMID- 10468504 TI - Frontal, midbrain and striatal dopaminergic function in early and advanced Parkinson's disease A 3D [(18)F]dopa-PET study. AB - We have studied focal changes in dopaminergic function throughout the brain volume in early and advanced Parkinson's disease by applying statistical parametric mapping (SPM) to 3D [(18)F]dopa-PET. Data from seven early hemi Parkinson's disease and seven advanced bilateral Parkinson's disease patients were compared with that from 12 normal controls. Parametric images of [(18)F]dopa influx rate constant (K(i)(o)) were generated for each subject from dynamic 3D [(18)F]dopa datasets and transformed into standard stereotactic space. Significant changes in mean voxel [(18)F]dopa K(i)(o) values between the normal control group and each Parkinson's disease group were localized with SPM. Conventional region of interest analysis was also applied to comparable regions on the untransformed image datasets. In early left hemi-Parkinson's disease, significant extrastriatal increases in [(18)F]dopa K(i)(o) were observed in the left anterior cingulate gyrus and the dorsal midbrain region (P < 0.05, corrected) along with decreases in striatal [(18)F]dopa K(i)(o). In advanced Parkinson's disease, significant extrastriatal decreases in [(18)F]dopa K(i)(o) were observed in the ventral and dorsal midbrain regions (P < 0.05, corrected). No significant changes in [(18)F]dopa K(i)(o) were observed in the anterior cingulate region. In a direct comparison between the early and late Parkinson's disease groups, we observed relative [(18)F]dopa K(i)(o) reductions in ventral and dorsal midbrain, and dorsal pontine regions along with striatal [(18)F]dopa K(i)(o) reductions. Similiar results were found with a region of interest approach, on non-transformed data, except for the focal midbrain [(18)F]dopa K(i)(o) increase seen in early Parkinson's disease. In conclusion, using SPM with [(18)F]dopa-PET, we have objectively localized changes in extrastriatal, pre synaptic dopaminergic function in Parkinson's disease. The significance of the increased dopaminergic activity of anterior cingulate in early Parkinson's disease remains unclear, but may be compensatory. The [(18)F]dopa signal in dorsal midbrain and pontine regions suggests that [(18)F]dopa is taken up by serotonergic and noradrenergic neurons which also degenerate in advanced Parkinson's disease. This suggests, therefore, that Parkinson's disease is a monoaminergic neurodegenerative disorder. PMID- 10468505 TI - Sensory processing in Parkinson's and Huntington's disease: investigations with 3D H(2)(15)O-PET. AB - There is conjoining experimental and clinical evidence supporting a fundamental role of the basal ganglia as a sensory analyser engaged in central somatosensory control. This study was aimed at investigating the functional anatomy of sensory processing in two clinical conditions characterized by basal ganglia dysfunction, i.e. Parkinson's and Huntington's disease. Based on previously recorded data of somatosensory evoked potentials, we expected deficient sensory-evoked activation in cortical areas that receive modulatory somatosensory input via the basal ganglia. Eight Parkinson's disease patients, eight Huntington's disease patients and eight healthy controls underwent repetitive H(2)(15)O-PET activation scans during two experimental conditions in random order: (i) continuous unilateral high-frequency vibratory stimulation applied to the immobilized metacarpal joint of the index finger and (ii) rest (no vibratory stimulus). In the control cohort, the activation pattern was lateralized to the side opposite to stimulus presentation, including cortical [primary sensory cortex (S1); secondary sensory cortex (S2)] and subcortical (globus pallidus, ventrolateral thalamus) regional cerebral blood flow (rCBF) increases (P < 0.001). Between-group comparisons (P < 0.01) of vibration-induced rCBF changes between patients and controls revealed differences in central sensory processing: (i) in Parkinson's disease, decreased activation of contralateral sensorimotor (S1/M1) and lateral premotor cortex, contralateral S2, contralateral posterior cingulate, bilateral prefrontal cortex (Brodmann area 10) and contralateral basal ganglia; (ii) in Huntington's disease, decreased activation of contralateral S2, parietal areas 39 and 40, and lingual gyrus, bilateral prefrontal cortex (Brodmann areas 8, 9, 10 and 44), S1 (trend only) and contralateral basal ganglia; (iii) in both clinical conditions relative enhanced activation of ipsilateral sensory cortical areas, notably caudal S1, S2 and insular cortex. Our data show that Parkinson's disease and Huntington's disease, beyond well-established deficits in central motor control, are characterized by abnormal cortical and subcortical activation on passive sensory stimulation. Furthermore, the finding that activation increases in ipsilateral sensory cortical areas may be interpreted as an indication of either altered central focusing and gating of sensory impulses, or enhanced compensatory recruitment of associative sensory areas in the presence of basal ganglia dysfunction. Altered sensory processing is thought to contribute to pertinent motor deficits in both conditions. PMID- 10468506 TI - Reduced basal ganglia blood flow and volume in pre-symptomatic, gene-tested persons at-risk for Huntington's disease. AB - The aim of this study was to examine basal ganglia volumes and regional cerebral blood flow in asymptomatic subjects at-risk for Huntington's disease who had undergone genetic testing. We determined which measures were the best 'markers' for the presence of the mutation and for the onset of symptoms. Twenty subjects who were Huntington's disease gene mutation-positive and 24 Huntington's disease gene mutation-negative participants, all of whom had a parent with genetically confirmed Huntington's disease, and were therefore 50% at-risk for inheriting the Huntington's disease gene mutation, were included in the study. To evaluate basal ganglia structure and function, MRI and single photon emission computed tomography (SPECT) were used. Quantitative measures of regional volumes and relative measures of regional perfusion were calculated. SPECT and MRI scans were co-registered so that MRI anatomy could be used accurately to place SPECT regions. Estimated years-to-onset in the mutation-positive subjects was calculated based on a regression formula that included gene (CAG)(n) repeat length and parental age of onset. Changes in imaging measures in relation to estimated years-to-onset were assessed. The imaging measure that was most affected in mutation-positive subjects was putamen volume. This was also the measure that correlated most strongly with approaching onset. In subjects >/=7 years from estimated onset age, the putamen volume measures were similar to those of the mutation-negative subjects. However, in subjects 90% discrimination from both the far-from-onset and the mutation-negative subjects. Caudate volume and bicaudate ratio also showed a significant decline in the close to-onset subjects, although to a lesser degree than putamen volume reductions. Furthermore, SPECT basal ganglia perfusion deficits were observed in mutation positive subjects. Imaging markers of neuropathological decline preceding clinical onset are important for assessing the effects of treatments aimed at slowing the course of Huntington's disease. The current study suggests that quantitative assessment of basal ganglia may provide a means to track early signs of decline in individuals with the Huntington's disease gene mutation prior to clinical onset. PMID- 10468507 TI - EEG-triggered functional MRI of interictal epileptiform activity in patients with partial seizures. AB - EEG-triggered functional MRI (fMRI) offers the potential to localize the generators of scalp EEG events, such as interictal epileptiform discharges, using a biological measurement as opposed to relying solely on modelling techniques. Although recent studies have demonstrated these possibilities in a small number of patients, wider application has been limited by concerns about patient safety, severe problems due to pulse-related artefact obscuring the EEG trace, and lack of reproducibility data. We have systematically studied and resolved the issues of patient safety and pulse artefact and now report the application of the technique in 24 experiments in 10 consecutive patients with localization-related epilepsy and frequent interictal epileptiform discharges (spikes or spike wave). At least two experiments were performed for each patient. In each experiment, 10- or 20-slice snapshot gradient-echo planar images were acquired approximately 3.5 s after a single typical epileptiform discharge (activation image) and in the absence of discharges (control image). Between 21 and 50 epileptiform discharges were sampled in each experiment. The significance of functional activation was tested using the t test at 95% confidence on a pixel-by-pixel basis. Six of the 10 patients showed reproducible focal changes of the blood oxygen level-dependent (BOLD) signal, which occurred in close spatial relationship to the maximum of the epileptiform discharges in the concurrent EEG. No reproducible focal BOLD signal changes were observed in the remaining four patients. In conclusion, EEG triggered fMRI is now a sufficiently developed technique to be more widely used in clinical studies, demonstrating that it can reproducibly localize the brain areas involved in the generation of spikes and spike wave in epilepsy patients with frequent interictal discharges. PMID- 10468508 TI - HLA-DPB1*0501-associated opticospinal multiple sclerosis: clinical, neuroimaging and immunogenetic studies. AB - In order to clarify the relationship between the clinical phenotype and the human leucocyte antigen (HLA) in multiple sclerosis in Asians, 93 Japanese patients with clinically definite multiple sclerosis underwent clinical MRI and HLA-DPB1 gene typing studies. According to a neurological examination, 29 patients were classified as opticospinal multiple sclerosis, 17 as spinal multiple sclerosis and 47 as Western type multiple sclerosis showing the involvement of multiple sites in the CNS including either the cerebrum, cerebellum or brainstem. The opticospinal multiple sclerosis showed a significantly higher age of onset, higher expanded disability status scale scores and higher CSF cell counts and protein content than the Western type multiple sclerosis. On brain and spinal cord MRI, the opticospinal multiple sclerosis showed a significantly lower number of brain lesions, but a higher frequency of gadolinium-enhancement of the optic nerve and a higher frequency of spinal cord atrophy than in Western type multiple sclerosis. The frequency of the HLA-DPB1*0501 allele was found to be significantly greater in opticospinal multiple sclerosis (93%) than in healthy controls (63%, corrected P value = 0.0091 and relative risk = 7.9), but not in Western type multiple sclerosis (66%) or spinal multiple sclerosis (82%). The marked differences in the clinical and MRI findings as well as in the immunogenetic backgrounds between the opticospinal multiple sclerosis and Western type multiple sclerosis together suggest that HLA-DPB1*0501-associated opticospinal multiple sclerosis is a distinct subtype of multiple sclerosis. PMID- 10468509 TI - The interleukin-1 type I receptor is expressed in human hypothalamus. AB - Several lines of evidence suggest that interleukin-1 (IL-1) acts directly on the central nervous system, probably within the hypothalamus, causing effects such as fever, activation of the immune response and sickness behaviour. IL-1 has also been shown to be involved in the aetiology of several neuronal diseases, including neurodegeneration, stroke and Alzheimer's disease. However, the question as to whether the full-length type I IL-1 receptor (IL-1RI) is expressed in the human hypothalamus has yet to be addressed. Using the polymerase chain reaction, we cloned a full-length cDNA encoding the human hypothalamic IL-1RI from human hypothalamic cDNA. The DNA sequence of the human hypothalamic receptor was identical to that of the human fibroblast IL-1RI. The IL-1RI receptor protein was detected in astrocytes of normal human hypothalamic brain sections using immunocytochemical techniques. To ascertain that the cloned receptor was functional, Chinese hamster ovary (CHO) cells were transfected with a plasmid vector containing the IL-1RI coding region. IL-1RI-mediated-signal transduction was assessed by microphysiometry and activation of p38 MAP (mitogen-activated protein) kinase. We report the first demonstration that both the type I IL-1 transcript and the protein are expressed in the human hypothalamus. The receptor was expressed in a stable CHO cell line, providing a tool with which to embark on a thorough analysis of the signalling mechanisms mediated by IL-1 via this receptor. PMID- 10468510 TI - The impact of different presenilin 1 andpresenilin 2 mutations on amyloid deposition, neurofibrillary changes and neuronal loss in the familial Alzheimer's disease brain: evidence for other phenotype-modifying factors. AB - To assess the influence of the presenilin 1 (PS1) and 2 (PS2) mutations on amyloid deposition, neurofibrillary tangle (NFT) formation and neuronal loss, we performed stereologically based counts in a high-order association cortex, the superior temporal sulcus, of 30 familial Alzheimer's disease cases carrying 10 different PS1 and PS2 mutations, 51 sporadic Alzheimer's disease cases and 33 non demented control subjects. All the PS1 and PS2 mutations assessed in this series led to enhanced deposition of total Abeta and Abeta(x-42/43) but not Abeta(x-40) senile plaques in the superior temporal sulcus when compared with brains from sporadic Alzheimer's disease patients. Some of the PS1 mutations studied (M139V, I143F, G209V, R269H, E280A), but not others, were also associated with faster rates of NFT formation and accelerated neuronal loss in the majority of the patients who harboured them when compared with sporadic Alzheimer's disease patients. In addition, our analysis showed that dramatic quantitative differences in clinical and neuropathological features can exist even among family members with the identical PS mutation. This suggests that further individual or pedigree genetic or epigenetic factors are likely to modulate PS phenotypes strongly. PMID- 10468511 TI - Interhemispheric asymmetries in the perception of unimanual and bimanual cutaneous stimuli. A study using transcranial magnetic stimulation. AB - Previous studies have shown that transcranial magnetic stimulation (TMS) of the sensorimotor cortex can induce a suppression of cutaneous perception from the fingers of the contralateral hand. In this work, 17 normal subjects were submitted to focal TMS of frontal and parietal scalp sites of each hemisphere. TMS was delivered at two interstimulus intervals (20 and 40 ms) following a cutaneous electrical stimulation of the first, third and fifth digits of either hand or both hands near the subjective threshold of perception. The aim of our study was to investigate whether TMS could detect an asymmetrical hemispheric specialization in the sensory perception of unimanual and bimanual, ipsilateral and contralateral sensory stimuli. At each interpulse interval, the right parietal cortex was significantly more sensitive to TMS interference with stimulus detection for both contralateral and ipsilateral stimuli compared with the left parietal cortex. These effects were mainly evident during bimanual discrimination tasks. Our results are indicative of an interhemispheric difference in the detection of cutaneous sensation, showing right hemispheric prevalence in the perception of contralateral as well as of ipsilateral stimuli. They provide neurophysiological support in normal humans to the clinical evidence which indicates that right hemisphere lesions can indeed produce deficits in the perception of ipsilateral sensory stimuli. PMID- 10468512 TI - Left frontal transcranial magnetic stimulation reduces contralesional extinction in patients with unilateral right brain damage. AB - It has been demonstrated previously that transcranial magnetic stimulation (TMS) of the sensorimotor cortex can induce transient suppression of the perception of cutaneous near-threshold stimuli from fingers of the contralateral hand in normal individuals. One explanation accounting for deficits in the exploration of contralateral space following a unilateral hemispheric lesion refers to a loss of the normal interhemispheric balance, with a resultant hyperactivation of the unaffected hemisphere due to the release of reciprocal inhibition by the affected one. In order to verify this hypothesis, we investigated the effects of a TMS induced transient dysfunction of the normal hemisphere upon contralateral tactile extinctions in two groups: (i) 14 right brain-damaged patients and (ii) 14 left brain-damaged control patients. Single-pulse TMS was delivered to frontal and parietal scalp sites of the unaffected hemisphere after an interval of 40 ms from an electrical unimanual or bimanual digit stimulation. In right brain-damaged patients, left frontal TMS significantly reduced the rate of contralateral extinctions compared with controls. After left parietal TMS, the number of extinctions was comparable to the baseline. This pattern of increased sensitivity to cutaneous stimulation ipsilateral to TMS was not observed in left brain damaged control patients. In this group, right hemisphere TMS did not significantly alter the recognition of bimanual stimuli delivered to the space contralateral to the lesion. The suggestion is made that extinctions produced by right brain damage may be dependent on a breakdown in the balance of hemispheric rivalry in directing spatial attention to contralateral hemispace, so that the unaffected hemisphere generates an unopposed orienting response to the side of the lesion. The mechanisms whereby the left frontal TMS transiently ameliorates these deficits may involve stimulus-induced removal of a left frontal-right parietal transcallosal inhibitory flow, although interactions at subcortical levels cannot be excluded. PMID- 10468513 TI - Locus-phenotype correlations in autosomal dominant pure hereditary spastic paraplegia. A clinical and molecular genetic study of 28 United Kingdom families. AB - This study aimed to describe the clinical phenotype of a large collection of families with autosomal dominant pure hereditary spastic paraplegia (ADPHSP), to examine the relative frequency of each of the three known ADPHSP genes within this population, to assess locus-phenotype correlation in ADPHSP and to ascertain whether there are clinical subgroups within genetically defined populations of ADPHSP families. We examined 306 family members, 144 affected, from 28 families with ADPHSP. Linkage analysis at the three known ADPHSP loci allowed us to categorize the families into three groups: (i) those families showing linkage to the chromosome 2 ADPHSP locus (seven families); (ii) those in which linkage to all known loci was excluded (five families); and (iii) those in which linkage results were inconclusive. There was a correlation between linkage group and clinical features, with chromosome 2-linked families having a later age at onset of symptoms (P = 0.001) and later age before commencing walking stick use (P = 0.007) than those families in which linkage to all known ADPHSP loci was excluded. There were no clinical differences between the families showing linkage to the chromosome 2 locus, but there were clinical differences between the families in which linkage to all of the known loci had been excluded (P < 0.0001). We conclude that the chromosome 2 ADPHSP gene is a frequent cause of ADPHSP in UK families, that the responsible gene has not yet been mapped in a significant proportion of families and that certain clinical features of ADPHSP, including age at onset, are at least in part determined by genetic locus. PMID- 10468514 TI - Changes in transmission in the pathway of heteronymous spinal recurrent inhibition from soleus to quadriceps motor neurons during movement in man. AB - H reflexes were induced in the human quadriceps muscle by electrical stimulation of the femoral nerve. The reflexes were conditioned by prior stimulation of the inferior soleus nerve. The conditioning stimulus produced an inhibition of long duration (>20 ms). The threshold of this inhibition was at zero soleus motor discharge and the inhibition scaled with soleus motor discharge. It was concluded that the inhibition was a heteronymous recurrent inhibition of quadriceps motor neurons mediated by Renshaw cells which had been activated by soleus motor neuron discharge. This recurrent inhibition declined during voluntary tonic contraction of the quadriceps, falling to zero at around one-third of maximum voluntary contraction. Antagonist contraction and weak co-contraction of the quadriceps and its antagonists did not lead to any significant change in recurrent inhibition. It is concluded that motor commands descending from the brain reduce heteronymous recurrent inhibition during isolated quadriceps muscle contraction, but to a much lesser extent during co-contraction. No evidence was obtained for any descending facilitation of heteronymous recurrent inhibition. PMID- 10468515 TI - Haemodynamic brain responses to acute pain in humans: sensory and attentional networks. AB - Turning attention towards or away from a painful heat stimulus is known to modify both the subjective intensity of pain and the cortical evoked potentials to noxious stimuli. Using PET, we investigated in 12 volunteers whether pain-related regional cerebral blood flow (rCBF) changes were also modulated by attention. High (mean 46.6 degrees C) or low (mean 39 degrees C) intensity thermal stimuli were applied to the hand under three attentional conditions: (i) attention directed towards the stimuli, (ii) attention diverted from the stimuli, and (iii) no task. Only the insular/second somatosensory cortices were found to respond whatever the attentional context and might, therefore, subserve the sensory discriminative dimension of pain (intensity coding). In parallel, other rCBF changes previously described as 'pain-related' appeared to depend essentially on the attentional context. Attention to the thermal stimulus involved a large network which was primarily right-sided, including prefrontal, posterior parietal, anterior cingulate cortices and thalamus. Anterior cingulate activity was not found to pertain to the intensity coding network but rather to the attentional neural activity triggered by pain. The attentional network disclosed in this study could be further subdivided into a non-specific arousal component, involving thalamic and upper brainstem regions, and a selective attention and orientating component including prefrontal, posterior parietal and cingulate cortices. A further effect observed in response to high intensity stimuli was a rCBF decrease within the somatosensory cortex ipsilateral to stimulation, which was considered to reflect contrast enhancing and/or anticipation processes. Attentional processes could possibly explain part of the variability observed in previous PET reports and should therefore be considered in further studies on pain in both normal subjects and patients with chronic pain. PMID- 10468516 TI - Training-induced brain plasticity in aphasia. AB - It has long been a matter of debate whether recovery from aphasia after left perisylvian lesions is mediated by the preserved left hemispheric language zones or by the homologous right hemisphere regions. Using PET, we investigated the short-term changes in the cortical network involved in language comprehension during recovery from aphasia. In 12 consecutive measurements of regional cerebral blood flow (rCBF), four patients with Wernicke's aphasia, caused by a posterior left middle cerebral artery infarction, were tested with a language comprehension task. Comprehension was estimated directly after each scan with a modified version of the Token Test. In the interval between the scans, the patients participated in brief, intense language comprehension training. A significant improvement in performance was observed in all patients. We correlated changes in blood flow measured during the language comprehension task with the scores achieved in the Token Test. The regions which best correlated with the training induced improvement in verbal comprehension were the posterior part of the right superior temporal gyrus and the left precuneus. This study supports the role of the right hemisphere in recovery from aphasia and demonstrates that the improvement in auditory comprehension induced by specific training is associated with functional brain reorganization. PMID- 10468517 TI - Stent-versus-stent equivalency trials : are some stents more equal than others? PMID- 10468518 TI - Angiotensin II induces LOX-1, the human endothelial receptor for oxidized low density lipoprotein. AB - BACKGROUND: Oxidatively modified LDL (oxLDL) plays an important role in the development of atherosclerosis. OxLDL effects, eg, foam cell formation, are mediated in part by the classic scavenger receptor, whereas other effects may involve the recently cloned endothelial oxLDL receptor, LOX-1 (lectinlike oxLDL receptor-1), which is distinct from macrophage scavenger receptors. Because the regulation of LOX-1 must still be defined, we investigated whether LOX-1 is regulated by the potentially proatherosclerotic stimulant angiotensin II (Ang II). METHODS AND RESULTS: Using competitive reverse transcription-polymerase chain reaction (RT-PCR), we quantified mRNA expression of LOX-1 in primary cultures of human umbilical vein endothelial cells (HUVECs). After treatment with Ang II for 3 hours (1 nmol/L to 1 micromol/L), LOX-1 mRNA was concentration dependently induced (from 6.9+/-1.4 to 23.1+/-5.5 relative units [RU] by 1 micromol/L Ang II; P<0.05). The angiotensin II type 1 (AT(1)) receptor antagonist losartan prevented this induction. Incubation of HUVECs with Ang II (100 nmol/L, 3 hours) induced LOX-1 protein expression (212+/-21% of control level; P<0. 01) and uptake of 1,1'-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate (DiI)-labeled oxLDL (209+/-17% of control level; P<0.05) by an AT(1)-dependent pathway, reaching its maximum after 24 hours (680+/-89%; P<0.05). In internal mammary artery biopsy samples from patients with or without ACE inhibitor treatment before coronary artery bypass surgery, LOX-1 mRNA was downregulated by ACE inhibition (6.4+/-2.0 versus 19.3+/-5. 9 RU; n=12 each; P<0.05). CONCLUSIONS: We conclude that LOX-1 is regulated by Ang II in vitro and in vivo, that induction of LOX-1 is mediated by the AT(1) receptor, and that repression of LOX 1 by long-term ACE inhibitor treatment may contribute to the antiatherosclerotic potential of this therapy. PMID- 10468519 TI - Process of progression of coronary artery lesions from mild or moderate stenosis to moderate or severe stenosis: A study based on four serial coronary arteriograms per year. AB - BACKGROUND: The process of progression in coronary artery disease is unknown. METHODS AND RESULTS: The subjects were 36 patients with 36 objective vessels with clinically significant progression of coronary artery disease (>/=15% per year) in whom 4 serial coronary arteriograms (CAGs) were performed at intervals of approximately 4 months in a 1-year period. The degree of progression of percent stenosis between each of 2 serial CAGs was classified as marked (M: >/=15%), slight (S: 5% to 14%), and no progression (N: <5%). From the pattern of progression, the 36 vessels were classified as 14 type 1 vessels with marked progression (N-->N-->M in 13 vessels and S-->S-->M in 1 vessel) and 22 type 2 vessels without marked progression (S-->S-->S in 18 vessels, N-->S-->S in 4). Percent stenosis at the first, second, third, and final CAGs was 44+/-14%, 46+/ 13%, 46+/-13%, and 88+/-10% (P<0.05 versus first CAG) in type 1 vessels and 44+/ 11%, 50+/-9%, 59+/-9%, and 67+/-9% in type 2 vessels (P<0.05 for second, third, and final CAGs versus first CAG). Type 1 vessels featured the sudden appearance of severe stenosis due to marked progression, angina pectoris, or myocardial infarction (71%) and Ambrose type II eccentric lesions indicating plaque rupture or thrombi (57%). Type 2 vessels featured continuous slight progression of stenosis with smooth vessel walls; angina pectoris (14%) occurred when the percent stenosis reached a severe level. An increase in serum C-reactive protein was observed only in the type 2 vessel group, which suggests a relation between continuous slight progression and inflammatory change. CONCLUSIONS: Two types of stenosis progression provide a new insight into the mechanism of coronary artery disease. PMID- 10468520 TI - Influence of pre-PTCA strategy and initial PTCA result in patients with multivessel disease: the Bypass Angioplasty Revascularization Investigation (BARI). AB - BACKGROUND: In PTCA patients with multivessel coronary artery disease, incomplete revascularization (IR) is the result of both pre-PTCA strategy and initial lesion outcome. The unique contribution of these components on long-term patient outcome is uncertain. METHODS AND RESULTS: From the Bypass Angioplasty Revascularization Investigation (BARI), 2047 patients who underwent first-time PTCA were evaluated. Before enrollment, all significant lesions were assessed by the PTCA operator for clinical importance and intention to dilate. Complete revascularization (CR) was defined as successful dilatation of all clinically relevant lesions. Planned CR was indicated in 65% of all patients. More lesions were intended for PTCA in these patients compared with those with planned IR (2.8 versus 2.1). Successful dilatation of all intended lesions occurred in 45% of patients with planned CR versus 56% with planned IR (P<0. 001). In multivariable analysis, planned IR (versus planned CR), initial lesions attempted (not all versus all intended lesions attempted), and initial lesion outcome (not all versus all attempted lesions successful) were unrelated to 5-year risk of cardiac death or death/myocardial infarction but were all independently related to risk of CABG. CONCLUSIONS: Overall, a pre-PTCA strategy of IR in BARI-like patients appears comparable to a strategy of CR except for a higher need for CABG. Whether the use of new devices may attenuate the elevated risk of CABG in patients with multivessel disease and planned IR remains to be determined. PMID- 10468521 TI - Influence of balloon pressure during stent placement in native coronary arteries on early and late angiographic and clinical outcome: A randomized evaluation of high-pressure inflation. AB - BACKGROUND: High-pressure dilatation is considered a better stent placement strategy, but this has not yet been proved by appropriately designed studies. The objective of this randomized trial was to assess the role of high-pressure dilatation in the early and late outcome of patients undergoing coronary stent placement. METHODS AND RESULTS: Consecutive patients with coronary stent placement were randomly assigned to high- (15 to 20 atm, 468 patients) or low- (8 to 13 atm, 466 patients) balloon-pressure dilatation. The primary end point of the study was the event-free survival at 1 year. Secondary end points were the incidence of stent thrombosis at 30 days and angiographic restenosis (>/=50% diameter stenosis) at 6 months. The incidence of stent thrombosis was 1.7% in the high-pressure and 1.9% in the low-pressure group (relative risk 0.89; 95% CI 0.30 to 2.56). During the first 30 days, although there was no significant difference in the incidence of Q-wave myocardial infarction, the incidence of non-Q-wave infarction was 6.4% in the high-pressure and 3.4% in the low-pressure group (relative risk 1. 87; 95% CI 1.02 to 3.42). The restenosis rate was 30.4% in the high-pressure and 31.4% in the low-pressure group (relative risk 0. 97; 95% CI 0.75 to 1.26). Event-free survival at 1 year was not significantly different between the groups, with 78.8% in high-pressure patients and 75.5% in patients assigned to low-pressure dilatation (hazard ratio 0.85; 95% CI 0.65 to 1.11). CONCLUSIONS: The systematic use of high-balloon-pressure inflation (15 to 20 atm) during coronary stent placement is not associated with any significant influence on the 1-year outcome of patients undergoing this intervention. PMID- 10468522 TI - Prognostication in 3-vessel coronary artery disease based on left ventricular ejection fraction during exercise : influence of coronary artery bypass grafting. AB - BACKGROUND: Previous data indicate that left ventricular ejection fraction (LVEF) provides prognostic information among patients with coronary artery disease (CAD), but the value of such testing specifically for defining benefits of coronary artery bypass grafting (CABG) may relate to severity of exercise inducible ischemia measured noninvasively before surgery. METHODS AND RESULTS: To determine the independent prognostic importance of preoperative ischemia severity for predicting outcomes of CABG among patients with extensive CAD, we monitored 167 stable patients with angiographically documented 3-vessel CAD (average follow up of 9 years in event-free patients) who previously had undergone rest and exercise radionuclide cineangiography. Their course was correlated with data obtained during initial radionuclide testing, coronary arteriography, and clinical evaluation at study entry. Fifty-two patients received medical treatment only, and 115 underwent CABG (44 early [1. CONCLUSIONS: Cytokine activation seems to play a significant pathogenetic role in both HIV-DCM and IDCM. In HIV-DCM patients, the state of immunodeficiency may favor the selection of viral variants of increased pathogenicity, influencing the clinical course of cardiomyopathy by enhancement of the inflammatory process. PMID- 10468525 TI - Moderate alcohol consumption and the risk of sudden cardiac death among US male physicians. AB - BACKGROUND: Individuals who consume high amounts of alcohol (>5 drinks/d) have increased risks of ventricular arrhythmia and sudden cardiac death (SCD). However, the relationship is less clear for drinkers of light-to-moderate amounts. METHODS AND RESULTS: We prospectively assessed whether light-to-moderate alcohol drinkers have a decreased risk of SCD among 21 537 male participants in the Physicians Health Study who were free of self-reported cardiovascular disease and provided complete information on alcohol intake at study entry. Over 12 years of follow-up, 141 SCDs were confirmed. After control for multiple confounders, men who consumed 2 to 4 drinks/wk (RR=0.40; 95% CI, 0.22 to 0.75; P=0.004) or 5 to 6 drinks/wk (RR=0.21; 95% CI, 0.08 to 0.56; P=0.002) at baseline had significantly reduced risks of SCD compared with those who rarely or never consumed alcohol. The relationship for SCD was U-shaped (P=0. 002), with the risk approaching unity at >/=2 drinks/d. In contrast, the relationship of alcohol intake and nonsudden CHD death was L-shaped or linear (P for trend=0.02). CONCLUSIONS: In these prospective data, men who consumed light-to-moderate amounts of alcohol (2 to 6 drinks/wk) had a significantly reduced risk of SCD compared with those who rarely or never consumed alcohol. PMID- 10468526 TI - Common carotid intima-media thickness and arterial stiffness: indicators of cardiovascular risk in high-risk patients. The SMART Study (Second Manifestations of ARTerial disease). AB - BACKGROUND: Common carotid intima-media thickness (IMT) and distensibility are markers of structural and functional vessel wall properties. Both parameters have been found in population-based studies to be associated with cardiovascular risk factors and prevalent cardiovascular disease. We investigated cross-sectionally whether IMT and distensibility are associated with cardiovascular risk in patients who already have vascular disease or atherosclerotic risk factors and evaluated the diagnostic ability of IMT and distensibility to discriminate between low- and high-risk patients. METHODS AND RESULTS: IMT and distensibility (change of diameter) of the left and right common carotid arteries were measured in the first 570 patients (537 for distensibility) enrolled in the Second Manifestations of ARTerial disease (SMART) study, a cohort study among patients with a manifestation of vascular disease or cardiovascular risk factors. Three risk scores were used to classify each patient's vascular risk. Areas under the curve (AUCs) of receiver-operating characteristic curves were calculated for IMT and distensibility after the patients were dichotomized on the median of the risk scores as the outcome. Risk scores increased nearly linearly with increasing IMT and decreasing distensibility. The AUCs for IMT predicting high-risk patients were 0.77, 0.73, and 0.77 based on the 3 risk scores. The AUCs for distensibility were 0. 65, 0.62, and 0.66. CONCLUSIONS: Common carotid IMT and distensibility are clear markers of cardiovascular risk in patients who already have vascular disease or atherosclerotic risk factors. IMT appears to discriminate between low- and high-risk patients better than distensibility. PMID- 10468527 TI - Ischemic preconditioning and the beta-adrenergic signal transduction pathway. AB - BACKGROUND: Previous studies from our laboratory showed cyclic increases in tissue cAMP during a multiple-cycle preconditioning (PC) protocol, followed by attenuated cAMP accumulation during sustained ischemia. The aim of this study was to determine whether ischemia-induced activation of the beta-adrenergic signaling pathway could act as a trigger in eliciting protection. METHODS AND RESULTS: Isolated perfused rat hearts were preconditioned by 3x5 minutes of global ischemia, interspersed by 5 minutes of reperfusion. beta-Adrenergic responsivity was assessed by measurement of tissue cAMP generation after beta-adrenergic agonist administration at the end of the PC protocol. Tissue cAMP, adenylyl cyclase, and protein kinase A (PKA) activities and beta-adrenergic receptor characteristics were assessed at different times. The role of cAMP generation in eliciting PC was studied by investigation of functional recovery during reperfusion after 25 minutes of global ischemia after (1) cAMP increases in the trigger period were prevented with the beta-adrenergic blocker alprenolol 7.5x10( 5) mol/L and (2) increases in cAMP were elicited by administration of forskolin 10(-7) and 10(-6) mol/L or isoproterenol 10(-8), 10(-7), and 10(-6) mol/L. Intermittent ischemia resulted in reduced beta-adrenergic responsivity at the end of the protocol, although B(max) and K(d) values of the beta-adrenergic receptor population and adenylyl cyclase and PKA activities were increased. Abolishment of cyclic increases in cAMP before sustained ischemia attenuated myocardial protection against ischemia, whereas agonists elicited protection. No clear correlation between protection and beta-adrenergic desensitization was observed. CONCLUSIONS: Ischemia-induced activation of the beta-adrenergic signaling pathway during preconditioning should also be considered a trigger in eliciting preconditioning. PMID- 10468528 TI - Requirement for protein kinase C in reactive oxygen species-induced apoptosis of vascular smooth muscle cells. AB - BACKGROUND: Vascular smooth muscle cell (VSMC) apoptosis is a component of a variety of cardiovascular diseases and may be related to reactive oxygen species (ROS). This study was designed to determine the role of protein kinase C (PKC) in ROS-induced VSMC apoptosis. METHODS AND RESULTS: Rat aortic VSMCs were exposed to H(2)O(2), and the nature of cell death was characterized in the absence or presence of different PKC inhibitors. The results demonstrate that exposure of VSMCs to H(2)O(2) led to a dose-dependent (25 to 100 micromol/L) and time dependent (peak at 2 minutes) activation of PKC. Among the PKC isoforms alpha, beta, delta, epsilon, and zeta, only PKC-alpha and PKC-epsilon were found to change their intracellular distribution on H(2)O(2) treatment. Apoptosis was the predominant form of cell death when PKC had been activated by H(2)O(2) alone or by H(2)O(2) in the presence of 50 nmol/L phorbol 12-myristate 13-acetate. In contrast, necrosis became the predominant form of cell death when PKC had been downregulated by prolonged exposure to 200 nmol/L phorbol 12,13-dibutyrate or inhibited by 50 nmol/L staurosporine, 100 nmol/L calphostin C, or 30 micromol/L H 7. In addition, caspase-3 was activated in H(2)O(2)-induced VSMC apoptosis but not when PKC was downregulated or inhibited. Inhibition of caspase-3 by 50 micromol/L Ac-DEVD-CHO could significantly attenuate H(2)O(2)-induced apoptosis and was not associated with the induction of necrosis. CONCLUSIONS: We conclude that in VSMCs, PKC converts the ROS-induced signals from necrotic cell death to the activation of an apoptotic cell death program. These data imply a novel and important role of PKC for the pathogenesis of such vascular diseases as atherosclerosis, restenosis, and hypertension. PMID- 10468529 TI - Effects of mutant and antisense RNA of phospholamban on SR Ca(2+)-ATPase activity and cardiac myocyte contractility. AB - BACKGROUND: The delayed cardiac relaxation in failing hearts has been attributed to a reduced activity of sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2). Phospholamban (PLB) inhibits SERCA2 activity and is therefore a potential target to improve the cardiac performance in heart failure. METHODS AND RESULTS: Mutants of PLB (Adv/mPLB) or antisense RNA of PLB (Adv/asPLB) was expressed in cardiac myocytes by recombinant adenovirus, and their effects on SERCA2 activity and myocyte contractility were studied. One mPLB, K3E/R14E, pentamerized with endogenous PLB in neonatal myocytes and resulted in a 45% increase in the affinity of SERCA2 for Ca(2+) and 27% faster diastolic Ca(2+) decline as determined by SR (45)Ca uptake assays and by indo 1-facilitated Ca(2+) transient measurement, respectively. Edge-detection analysis of adult myocyte contractility showed a 74% increase in fractional shortening, accompanied by 115% increase in velocity of relengthening and 25% decrease in time to half-maximal relengthening. In parallel, infection of neonatal cardiac myocytes by Adv/asPLB decreased the endogenous PLB level by 54%, which was associated with a 35% increase in Ca(2+) affinity of SERCA2 and 21% faster diastolic Ca(2+) decline. However, in adult cardiac myocytes, Adv/asPLB failed to significantly alter the endogenous PLB level, the SERCA2 activity, or most of the contractile parameters. CONCLUSIONS: K3E/R14E is a dominant negative mutant of PLB that disrupts the structural integrity and function of the endogenous PLB and consequently enhances SERCA2 activity and myocyte contractility. In neonatal myocytes, the decrease in steady state abundance of PLB by asPLB also leads to increased SERCA2 activity. PMID- 10468530 TI - Effects of chronic heparin administration on coronary vascular adaptation to hypertension and ventricular hypertrophy in sheep. AB - BACKGROUND: Hypertension decreases myocardial perfusion capacity in adults for several reasons, including insufficient coronary angiogenesis with left ventricular (LV) hypertrophy, arteriolar hypertrophy, and altered vasomotion. Heparin influences growth factors that promote angiogenesis and vasodilation and inhibit arteriolar wall thickening. METHODS AND RESULTS: Adult sheep were given heparin 200 U/kg body wt SC twice daily throughout 6 weeks of LV and coronary hypertension from a progressively constricted ascending aortic band (n=14). They were compared with untreated sheep with (n=13) and without (n=13) aortic stenosis. After 6 weeks, maximum myocardial perfusion was measured during adenosine infusion in the conscious state by the microsphere method. Sheep with aortic stenosis had less maximum coronary flow per gram, less conductance reserve, and thicker arteriolar walls in the LV and nonhypertrophied right ventricle. Capillary density decreased in the LV endomyocardium and remained unchanged in the right ventricle. Heparin-treated sheep had significant partial normalization of coronary conductance reserve and maximum perfusion in both ventricles and capillary density in the LV endomyocardium. Arteriolar wall thickness was unchanged. Compared with untreated sheep with aortic stenosis, in heparin-treated sheep LV FGF-2 protein increased 2-fold, whereas FGF-2 mRNA remained unchanged. VEGF mRNA and protein increased 3-fold and 1.4-fold, respectively, whereas TGF-beta(1) mRNA declined 3-fold. CONCLUSIONS: Heparin administration during LV hypertension increases heparin-binding angiogenic factors FGF-2 and VEGF in the LV and ameliorates decreases in LV perfusion capacity and capillary density. PMID- 10468531 TI - Primary prevention of coronary heart disease: integrating risk assessment with intervention. PMID- 10468532 TI - Mechanisms and models in heart failure: A combinatorial approach. PMID- 10468533 TI - Images in cardiovascular medicine. Cerebral arterial embolism from a protruding atheroma of the aortic arch after a nonpenetrating chest trauma. PMID- 10468534 TI - Amlodipine releases nitric oxide from canine coronary microvessels: An unexpected mechanism of action of a calcium channel-blocking agent. PMID- 10468535 TI - 1999 update: ACC/AHA Guidelines for the Management of Patients With Acute Myocardial Infarction: Executive Summary and Recommendations: A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee on Management of Acute Myocardial Infarction). PMID- 10468536 TI - Pleural effusion on transthoracic echocardiogram. PMID- 10468537 TI - Physiological reactions of nitric oxide and hemoglobin: a radical rethink. PMID- 10468538 TI - Folding and binding cascades: shifts in energy landscapes. PMID- 10468539 TI - Gene therapy for the hemophilias. PMID- 10468541 TI - SINEs of the perfect character. PMID- 10468540 TI - The education of a brain transplant. PMID- 10468542 TI - Introduction to Frontiers of Science. PMID- 10468543 TI - Quantum conductors in a plane. PMID- 10468544 TI - Location, location, location: the many addresses of memory formation. AB - Memory formation, like real estate, can be summarized succinctly-location, location, location. It is an emergent property involving different anatomical regions in the brain, sets of neuronal circuits, and cellular and molecular interactions between and within those neurons. At each of these levels of description, location continues to be a major organizing principle guiding researchers. The difficulty in the field is the integration of information between the various levels of analyses, and it is proposed that molecular reporters may help to fill that void. PMID- 10468545 TI - Global climate change. AB - Most of the last 100,000 years or longer has been characterized by large, abrupt, regional-to-global climate changes. Agriculture and industry have developed during anomalously stable climatic conditions. New, high-resolution analyses of sediment cores using multiproxy and physically based transfer functions allow increasingly confident interpretation of these past changes as having been caused by "band jumps" between modes of operation of the climate system. Recurrence of such band jumps is possible and might be affected by human activities. PMID- 10468547 TI - Frontiers of finance: evolution and efficient markets. AB - In this review article, we explore several recent advances in the quantitative modeling of financial markets. We begin with the Efficient Markets Hypothesis and describe how this controversial idea has stimulated a number of new directions of research, some focusing on more elaborate mathematical models that are capable of rationalizing the empirical facts, others taking a completely different tack in rejecting rationality altogether. One of the most promising directions is to view financial markets from a biological perspective and, specifically, within an evolutionary framework in which markets, instruments, institutions, and investors interact and evolve dynamically according to the "law" of economic selection. Under this view, financial agents compete and adapt, but they do not necessarily do so in an optimal fashion. Evolutionary and ecological models of financial markets is truly a new frontier whose exploration has just begun. PMID- 10468546 TI - Amyloid diseases: abnormal protein aggregation in neurodegeneration. PMID- 10468548 TI - Black holes in the milky way galaxy. AB - Extremely strong observational evidence has recently been found for the presence of black holes orbiting a few relatively normal stars in our Milky Way Galaxy and also at the centers of some galaxies. The former generally have masses of 4-16 times the mass of the sun, whereas the latter are "supermassive black holes" with millions to billions of solar masses. The evidence for a supermassive black hole in the center of our galaxy is especially strong. PMID- 10468549 TI - Morphological innovation and developmental genetics. AB - How do the actions of individual genes contribute to the complex morphologies of animals and plants? How widespread are these genes taxonomically? How many genes are involved in the morphological differences observed between species, and can we identify them? To what extent can empirical data and theory be reconciled? We provide an overview of some recent attempts to answer these questions, answers that have taken us to the threshold of understanding the mechanistic basis and evolutionary factors that underlie morphological innovation. PMID- 10468550 TI - The maximal affinity of ligands. AB - We explore the question of what are the best ligands for macromolecular targets. A survey of experimental data on a large number of the strongest-binding ligands indicates that the free energy of binding increases with the number of nonhydrogen atoms with an initial slope of approximately -1.5 kcal/mol (1 cal = 4.18 J) per atom. For ligands that contain more than 15 nonhydrogen atoms, the free energy of binding increases very little with relative molecular mass. This nonlinearity is largely ascribed to nonthermodynamic factors. An analysis of the dominant interactions suggests that van der Waals interactions and hydrophobic effects provide a reasonable basis for understanding binding affinities across the entire set of ligands. Interesting outliers that bind unusually strongly on a per atom basis include metal ions, covalently attached ligands, and a few well known complexes such as biotin-avidin. PMID- 10468551 TI - Electrochemical preparation of tris(tert-butyldimethylsilyl)cyclopropene and its hydride abstraction to tris(tert-butyldimethylsilyl)cyclopropenium tetrafluoroborate. AB - Electrochemical reductive tert-butyldimethylsilylation of tetrachlorocyclopropene to 1,2,3-tris(tert-butyldimethylsilyl)cyclopropene, a potential strained precursor for Diels-Alder and related cycloaddition reactions, is described. By hydride abstraction with nitrosonium tetrafluoroborate, 1,2,3-tris(tert butyldimethylsilyl)cyclopropene is ionized quantitatively to Huckeloid 2pi aromatic tris(tert-butyldimethylsilyl)cyclopropenium tetrafluoroborate. PMID- 10468552 TI - An isotopic approach for understanding the CH(3)Br budget of the atmosphere. AB - The atmospheric budget of methyl bromide (CH(3)Br), an ozone-depleting gas, is highly uncertain, because it has complex sources and sinks. Although oceans, biomass burning, and industrial production are identified as the major sources, the fraction of CH(3)Br that is contributed by each source is not well known. A mass-balance approach that exploits differences in the carbon isotopic signature (delta(13)C) of CH(3)Br sources and sinks may provide a means of reducing uncertainties in the atmospheric budget. This approach depends on the distinctiveness of industrially produced methyl bromide. Our delta(13)C measurements of industrial CH(3)Br from the three largest manufacturers worldwide yield a weighted average of -54.4 per thousand relative to the Peedee Belemnite standard. This result suggests that industrial CH(3)Br is isotopically distinct and that the carbon isotopic composition of atmospheric CH(3)Br may indicate what fraction of atmospheric CH(3)Br is anthropogenic. PMID- 10468553 TI - Highly efficient endonucleolytic cleavage of RNA by a Cys(2)His(2) zinc-finger peptide. AB - We have identified a 30-aa peptide that efficiently cleaves single-stranded RNA. The peptide sequence corresponds to a single zinc finger of the human male associated ZFY protein; a transcription factor belonging to the Cys(2)His(2) family of zinc-finger proteins. RNA cleavage was observed only in the absence of zinc. Coordination with zinc resulted in complete loss of ribonuclease activity. The ribonuclease active structure was determined to be a homodimeric form of the peptide. Dimerization of the peptide occurred through a single intermolecular disulfide between two of the four cystines. The observed hydrolytic activity was single-stranded RNA-specific. Single-stranded DNA, double-stranded RNA and DNA, and 2'-methoxy-modified sequences were not degraded by the peptide. The peptide specifically cleaved pyrimidines within single-stranded RNA and the dinucleotide sequence 5'-pyr-A-3' was preferred. The RNA cleavage products consisted of a 3' phosphate and 5' hydroxyl. The initial rates of cleavage (V(0)) observed for the finger peptide were comparable to rates observed for human ribonucleases, and the catalytic rate (K(cat)) was comparable to rates observed for the group II intron rybozymes. The pH profile exhibited by the peptide is characteristic of general acid-base catalytic mechanisms observed with other ribonucleases. These observations raise interesting questions about the potential biological roles of zinc-finger proteins. PMID- 10468554 TI - Chip-based genotyping by mass spectrometry. AB - Silicon chips with immobilized target DNAs were used for accurate genotyping by mass spectrometry. Genomic DNAs were amplified with PCR, and the amplified products were covalently attached to chip wells via N-succinimidyl (4 iodoacetyl)aminobenzoate (SIAB) chemistry. Primer annealing, extension, and termination were performed on a 1-microl scale directly in the chip wells in parallel. Diagnostic products thus generated were detected in situ by using matrix-assisted laser desorption ionization mass spectrometry. This miniaturized method has the potential for accurate, high-throughput, low-cost identification of genetic variations. PMID- 10468555 TI - Pathways for proton release during ubihydroquinone oxidation by the bc(1) complex. AB - Quinol oxidation by the bc(1) complex of Rhodobacter sphaeroides occurs from an enzyme-substrate complex formed between quinol bound at the Q(o) site and the iron-sulfur protein (ISP) docked at an interface on cytochrome b. From the structure of the stigmatellin-containing mitochondrial complex, we suggest that hydrogen bonds to the two quinol hydroxyl groups, from Glu-272 of cytochrome b and His-161 of the ISP, help to stabilize the enzyme-substrate complex and aid proton release. Reduction of the oxidized ISP involves H transfer from quinol. Release of the proton occurs when the acceptor chain reoxidizes the reduced ISP, after domain movement to an interface on cytochrome c(1). Effects of mutations to the ISP that change the redox potential and/or the pK on the oxidized form support this mechanism. Structures for the complex in the presence of inhibitors show two different orientations of Glu-272. In stigmatellin-containing crystals, the side chain points into the site, to hydrogen bond with a ring hydroxyl, while His-161 hydrogen bonds to the carbonyl group. In the native structure, or crystals containing myxothiazol or beta-methoxyacrylate-type inhibitors, the Glu 272 side chain is rotated to point out of the site, to the surface of an external aqueous channel. Effects of mutation at this residue suggest that this group is involved in ligation of stigmatellin and quinol, but not quinone, and that the carboxylate function is essential for rapid turnover. H(+) transfer from semiquinone to the carboxylate side chain and rotation to the position found in the myxothiazol structure provide a pathway for release of the second proton. PMID- 10468557 TI - RNA aptamers as effective protein antagonists in a multicellular organism. AB - RNA aptamers selected against proteins can be used to modulate specific protein function. Expression of such reagents in cells and whole organisms could provide a means of dissecting and controlling molecular mechanisms in vivo. We demonstrate that Drosophila B52 protein can be specifically inhibited in vitro and in vivo by a multivalent RNA aptamer. This inhibitory aptamer RNA binds B52 avidly and inhibits B52-stimulated pre-mRNA splicing. It can be expressed in cultured cells and whole animals in a stable form that accumulates up to 10% of total mRNA. It binds B52 in vivo and suppresses all phenotypes caused by B52 overexpression. The strategies presented here should prove general in design and expression of functional and therapeutic RNAs. PMID- 10468556 TI - Lamivudine (3TC) resistance in HIV-1 reverse transcriptase involves steric hindrance with beta-branched amino acids. AB - An important component of triple-drug anti-AIDS therapy is 2', 3'-dideoxy-3' thiacytidine (3TC, lamivudine). Single mutations at residue 184 of the reverse transcriptase (RT) in HIV cause high-level resistance to 3TC and contribute to the failure of anti-AIDS combination therapy. We have determined crystal structures of the 3TC-resistant mutant HIV-1 RT (M184I) in both the presence and absence of a DNA/DNA template-primer. In the absence of a DNA substrate, the wild type and mutant structures are very similar. However, comparison of crystal structures of M184I mutant and wild-type HIV-1 RT with and without DNA reveals repositioning of the template-primer in the M184I/DNA binary complex and other smaller changes in residues in the dNTP-binding site. On the basis of these structural results, we developed a model that explains the ability of the 3TC resistant mutant M184I to incorporate dNTPs but not the nucleotide analog 3TCTP. In this model, steric hindrance is expected for NRTIs with beta- or L- ring configurations, as with the enantiomer of 3TC that is used in therapy. Steric conflict between the oxathiolane ring of 3TCTP and the side chain of beta branched amino acids (Val, Ile, Thr) at position 184 perturbs inhibitor binding, leading to a reduction in incorporation of the analog. The model can also explain the 3TC resistance of analogous hepatitis B polymerase mutants. Repositioning of the template-primer as observed in the binary complex (M184I/DNA) may also occur in the catalytic ternary complex (M184I/DNA/3TCTP) and contribute to 3TC resistance by interfering with the formation of a catalytically competent closed complex. PMID- 10468558 TI - Purification, molecular cloning, and expression of 2-hydroxyphytanoyl-CoA lyase, a peroxisomal thiamine pyrophosphate-dependent enzyme that catalyzes the carbon carbon bond cleavage during alpha-oxidation of 3-methyl-branched fatty acids. AB - In the third step of the alpha-oxidation of 3-methyl-branched fatty acids such as phytanic acid, a 2-hydroxy-3-methylacyl-CoA is cleaved into formyl-CoA and a 2 methyl-branched fatty aldehyde. The cleavage enzyme was purified from the matrix protein fraction of rat liver peroxisomes and identified as a protein made up of four identical subunits of 63 kDa. Its activity proved to depend on Mg(2+) and thiamine pyrophosphate, a hitherto unrecognized cofactor of alpha-oxidation. Formyl-CoA and 2-methylpentadecanal were identified as reaction products when the purified enzyme was incubated with 2-hydroxy-3-methylhexadecanoyl-CoA as the substrate. Hence the enzyme catalyzes a carbon-carbon cleavage, and we propose calling it 2-hydroxyphytanoyl-CoA lyase. Sequences derived from tryptic peptides of the purified rat protein were used as queries to recover human expressed sequence tags from the databases. The composite cDNA sequence of the human lyase contained an ORF of 1,734 bases that encodes a polypeptide with a calculated molecular mass of 63,732 Da. Recombinant human protein, expressed in mammalian cells, exhibited lyase activity. The lyase displayed homology to a putative Caenorhabditis elegans protein that resembles bacterial oxalyl-CoA decarboxylases. Similarly to the decarboxylases, a thiamine pyrophosphate-binding consensus domain was present in the C-terminal part of the lyase. Although no peroxisome targeting signal, neither 1 nor 2, was apparent, transfection experiments with constructs encoding green fluorescent protein fused to the full length lyase or its C-terminal pentapeptide indicated that the C terminus of the lyase represents a peroxisome targeting signal 1 variant. PMID- 10468559 TI - New routes for lignin biosynthesis defined by biochemical characterization of recombinant ferulate 5-hydroxylase, a multifunctional cytochrome P450-dependent monooxygenase. AB - The enzymes and genes of the lignin biosynthetic pathway have been studied for several decades, but the gene encoding ferulate 5-hydroxylase (F5H) was cloned only 3 years ago by T-DNA tagging in Arabidopsis. To characterize the enzyme in detail, we have expressed F5H in yeast. According to current models of the phenylpropanoid pathway, F5H catalyzes the hydroxylation of ferulate to 5 hydroxyferulate; however, our studies indicate that the enzyme also uses coniferaldehyde and coniferyl alcohol as substrates. Unexpectedly, the K(m) values measured for the latter two substrates are three orders of magnitude lower than that measured for ferulic acid, suggesting that in lignifying tissues, syringyl monomers may be derived from their guaiacyl counterparts by hydroxylation and subsequent methylation. Thus, F5H may function later in the lignin biosynthetic pathway than was originally proposed. To further test this model, recombinant F5H was incubated together with ferulic acid, coniferaldehyde, or coniferyl alcohol in the presence of native or recombinant Arabidopsis caffeic acid/5-hydroxyferulic acid O-methyltransferase and [(14)C]S-adenosylmethionine. In all cases, the corresponding radiolabeled sinapyl derivatives were synthesized, indicating that the necessary enzymes required for this pathway are present in Arabidopsis. Taken together, these data suggest that the previously accepted pathway for lignin biosynthesis is likely to be incorrect. PMID- 10468560 TI - The beta-globin promoter is important for recruitment of erythroid Kruppel-like factor to the locus control region in erythroid cells. AB - Erythroid Kruppel-like factor (EKLF), which binds to the CACCC box in the beta globin promoter, is required for the expression of the beta-globin gene in adult erythroid cells. It was recently demonstrated that EKLF is also required for the activity of the beta-globin locus control region (LCR) 5'HS3. Some evidence suggests that the LCR and the beta-globin promoter interact in adult erythroid cells, and the network of protein-protein interactions that exists between these two elements may regulate how EKLF is recruited to the LCR. In this report, we use the PIN*POINT assay to study the role of the promoter on the recruitment of EKLF to 5'HS2 and 5'HS3 of the LCR. We find that recruitment of EKLF to 5'HS2 requires the TATA box, but recruitment to 5'HS3 depends on the CACCC and TATA boxes of the beta-globin promoter. Furthermore, recruitment of EKLF to 5'HS3 only occurred in beta-globin-expressing murine erythroid leukemia cells, whereas recruitment of EKLF to 5'HS2 occurred in both gamma-globin-expressing K562 cells and murine erythroid leukemia cells. Unlike EKLF, Sp1, which also binds to CACCC boxes, is not recruited to 5'HS3. We have also examined how one 5'HS affects the recruitment of EKLF to another 5'HS. We have found that the recruitment of EKLF to 5'HS3 depends on the presence of 5'HS2 in cis, but the recruitment to 5'HS2 does not depend on 5'HS3. Based on these results, we present a model that illustrates how EKLF may be recruited to the beta-globin locus. PMID- 10468561 TI - Mutational analysis of a higher plant antenna protein provides identification of chromophores bound into multiple sites. AB - The chromophore-binding properties of the higher plant light-harvesting protein CP29 have been studied by using site-directed mutagenesis of pigment-binding residues. Overexpression of the apoproteins in bacteria was followed by reconstitution in vitro with purified pigments, thus obtaining a family of mutant CP29 proteins lacking individual chromophore-binding sites. Biochemical characterization allowed identification of the eight porphyrins and two xanthophyll-binding sites. It is shown that the four porphyrin-binding sites (A1, A2, A4, and A5) situated in the central, twofold-symmetrical domain of the protein are selective for Chl-a, whereas the four peripheral sites (A3, B3, B5, and B6) have mixed Chl-a-Chl-b specificity. Within a site, porphyrin coordination by glutamine increases affinity for Chl-b as compared with glutamate. Xanthophyll site L1 is occupied by lutein, whereas site L2 can bind violaxanthin or neoxanthin. The protein is relatively stable when site L2 site is empty, suggesting that xanthophylls can be exchanged during operation of xanthophyll cycle-dependent photoprotection mechanism. Differential absorption spectroscopy allowed determination of transition energy levels for individual chromophores, thus opening the way to calculation of energy-transfer rates between Chl in higher plant antenna proteins. PMID- 10468562 TI - Crystal structure of triosephosphate isomerase from Trypanosoma cruzi in hexane. AB - To gain insight into the mechanisms of enzyme catalysis in organic solvents, the x-ray structure of some monomeric enzymes in organic solvents was determined. However, it remained to be explored whether the structure of oligomeric proteins is also amenable to such analysis. The field acquired new perspectives when it was proposed that the x-ray structure of enzymes in nonaqueous media could reveal binding sites for organic solvents that in principle could represent the starting point for drug design. Here, a crystal of the dimeric enzyme triosephosphate isomerase from the pathogenic parasite Trypanosoma cruzi was soaked and diffracted in hexane and its structure solved at 2-A resolution. Its overall structure and the dimer interface were not altered by hexane. However, there were differences in the orientation of the side chains of several amino acids, including that of the catalytic Glu-168 in one of the monomers. No hexane molecules were detected in the active site or in the dimer interface. However, three hexane molecules were identified on the surface of the protein at sites, which in the native crystal did not have water molecules. The number of water molecules in the hexane structure was higher than in the native crystal. Two hexanes localized at <4 A from residues that form the dimer interface; they were in close proximity to a site that has been considered a potential target for drug design. PMID- 10468563 TI - Protein synthesis by native chemical ligation: expanded scope by using straightforward methodology. AB - The total chemical synthesis of proteins has great potential for increasing our understanding of the molecular basis of protein function. The introduction of native chemical ligation techniques to join unprotected peptides next to a cysteine residue has greatly facilitated the synthesis of proteins of moderate size. Here, we describe a straightforward methodology that has enabled us to rapidly analyze the compatibility of the native chemical ligation strategy for X Cys ligation sites, where X is any of the 20 naturally occurring amino acids. The simplified methodology avoids the necessity of specific amino acid thioester linkers or alkylation of C-terminal thioacid peptides. Experiments using matrix assisted laser-desorption ionization MS analysis of combinatorial ligations of LYRAX-C-terminal thioester peptides to the peptide CRANK show that all 20 amino acids are suitable for ligation, with Val, Ile, and Pro representing less favorable choices because of slow ligation rates. To illustrate the method's utility, two 124-aa proteins were manually synthesized by using a three-step, four-piece ligation to yield a fully active human secretory phospholipase A(2) and a catalytically inactive analog. The combination of flexibility in design with general access because of simplified methodology broadens the applicability and versatility of chemical protein synthesis. PMID- 10468564 TI - Protein prenylation in spinach chloroplasts. AB - Protein prenylation in plants was studied by in vivo metabolic (3)H-mevalonate labeling in combination with a range of protein synthesis inhibitors. In spinach cotyledons, this posttranslational protein modification was found to be divided into two categories, one representing the conventional prenylation involving farnesyl and geranylgeranyl groups bound to cysteine residues via thioether linkages. This category revealed a similar pattern of prenylated proteins to that observed in mammalian cells and depends on nuclear gene expression. The other category was shown to represent a type of prenylation confined to chloroplasts. It depends on plastid gene expression and does not involve a thioether bond. The modifying isoprenoid could be released from the chloroplastic polypeptides by alkaline treatment and was identified as phytol upon GC-MS analysis. The phytol could readily be derived from all-trans-[(3)H]farnesol, which, like all-trans [(3)H]geranylgeraniol, was taken up by the cotyledons, resulting in incorporation of radiolabel into proteins. PMID- 10468565 TI - Induction of apoptosis by adenovirus E4orf4 protein is specific to transformed cells and requires an interaction with protein phosphatase 2A. AB - We previously have shown that adenovirus type 5 E4orf4 protein associates with protein phosphatase 2A (PP2A) and induces apoptosis in transformed cells in a p53 independent manner. Here we show that the interaction between E4orf4 and PP2A is required for induction of apoptosis by the viral protein. This conclusion is supported by a mutation analysis of E4orf4 protein, showing a correlation between the ability to bind PP2A and to induce apoptosis, and by the observation that transfection of an antisense construct of the PP2A-B55 subunit reduces expression of the PP2A-B55 subunit and inhibits induction of apoptosis by E4orf4, but not by p53. The mutant analysis also indicates that even a low level of interaction with PP2A is sufficient to initiate the E4orf4 apoptotic pathway. In addition, E4orf4 inhibits cellular transformation by various oncogenes, and this function is coupled to its ability to induce apoptosis. Furthermore, expression of oncogenes in primary cell cultures sensitizes these cells to induction of apoptosis by E4orf4. Our results suggest that E4orf4 is a potentially useful tool for cancer gene therapy. PMID- 10468566 TI - Horseshoe crab acetyl group-recognizing lectins involved in innate immunity are structurally related to fibrinogen. AB - We have characterized and cloned newly isolated lectins from hemolymph plasma of the horseshoe crab Tachypleus tridentatus, which we named tachylectins 5A and 5B (TLs-5). TLs-5 agglutinated all types of human erythrocytes and Gram-positive and Gram-negative bacteria. TLs-5 specifically recognize acetyl group-containing substances including noncarbohydrates; the acetyl group is required and is sufficient for recognition. TLs-5 enhanced the antimicrobial activity of a horseshoe crab-derived big defensin. cDNA sequences of TLs-5 indicated that they consist of a short N-terminal Cys-containing segment and a C-terminal fibrinogen like domain with the highest sequence identity (51%) to that of mammalian ficolins. TLs-5, however, lack the collagenous domain found in a kind of "bouquet arrangement" of ficolins and collectins. Electron microscopy revealed that TLs-5 form two- to four-bladed propeller structures. The horseshoe crab is equipped with a unique functional homologue of vertebrate fibrinogen, coagulogen, as the target protein of the clotting cascade. Our observations clearly show that the horseshoe crab has fibrinogen-related molecules in hemolymph plasma and that they function as nonself-recognizing lectins. An ancestor of fibrinogen may have functioned as a nonself-recognizing protein. PMID- 10468567 TI - In vivo transcription factor recruitment during thyroid hormone receptor-mediated activation. AB - Thyroid hormone receptor (TR) can act as both a transcriptional activator and a silencer. Optimal activation by TR requires synergism with activator(s) bound to the promoter (promoter proximal activator). It is thought that liganded TR either helps to recruit preinitiation complexes (PIC) to the promoter or activates the PIC already recruited. However, the studies analyzing the TR action on the PIC formation were done in vitro and, therefore, it is not clear how relevant they are to the in vivo TR action. For example, in vivo, the TR can act from distances equal to or greater than a kilobase from the promoter, but such distant effect is not reproducible in vitro. In this study, we used the PIN*POINT (ProteIN POsition Identification with Nuclease Tail) assay to define the molecular mechanism of TR action on transcription from the thymidine kinase promoter in the cellular context. We demonstrate that the recruitment of promoter-proximal activator Sp1, and the components of the basal transcription factors such as TBP, TFIIB, and Cdk7, is enhanced with thyroid hormone activation. Our results suggest that DNA forms a loop with TR-mediated activation to accommodate interactions between the liganded TR complex and the complex formed on the promoter. We also show that Sp1 bound to the promoter is essential for the DNA looping and recruitment of basal transcription factors such as TFIIB and Cdk7 but not for recruitment of TBP. On the basis of these findings, we present a model that illustrates the molecular mechanism of TR-mediated activation in vivo. PMID- 10468568 TI - Contribution of glucose transport to the control of the glycolytic flux in Trypanosoma brucei. AB - The rate of glucose transport across the plasma membrane of the bloodstream form of Trypanosoma brucei was modulated by titration of the hexose transporter with the inhibitor phloretin, and the effect on the glycolytic flux was measured. A rapid glucose uptake assay was developed to measure the transport activity independently of the glycolytic flux. Phloretin proved a competitive inhibitor. When the effect of the intracellular glucose concentration on the inhibition was taken into account, the flux control coefficient of the glucose transporter was between 0.3 and 0.5 at 5 mM glucose. Because the flux control coefficients of all steps in a metabolic pathway sum to 1, this result proves that glucose transport is not the rate-limiting step of trypanosome glycolysis. Under physiological conditions, transport shares the control with other steps. At glucose concentrations much lower than physiological, the glucose carrier assumed all control, in close agreement with model predictions. PMID- 10468569 TI - Heightened sensitivity of a lattice of membrane receptors. AB - Receptor proteins in both eukaryotic and prokaryotic cells have been found to form two-dimensional clusters in the plasma membrane. In this study, we examine the proposition that such clusters might show coordinated responses because of the spread of conformational states from one receptor to its neighbors. A Monte Carlo simulation was developed in which receptors flipped in probabilistic fashion between an active and an inactive state. Conformational energies depended on (i) ligand binding, (ii) a chemical modification of the receptor conferring adaptation, and (iii) the activity of neighboring receptors. Rate constants were based on data from known biological receptors, especially the bacterial Tar receptor, and on theoretical constraints derived from an analogous Ising model. The simulated system showed a greatly enhanced sensitivity to external signals compared with a corresponding set of uncoupled receptors and was operational over a much wider range of ambient concentrations. These and other properties should make a lattice of conformationally coupled receptors ideally suited to act as a "nose" by which a cell can detect and respond to extracellular stimuli. PMID- 10468570 TI - Polymerization and mechanical properties of single RecA-DNA filaments. AB - The polymerization of individual RecA-DNA filaments, containing either single stranded or double-stranded DNA, was followed in real time, and their mechanical properties were characterized with force-measuring laser tweezers. It was found that the stretch modulus of a filament is dominated by its (central) DNA component, while its bending rigidity is controlled by its (eccentric) protein component. The longitudinal stiffness of DNA increases 6- to 12-fold when the DNA is contained in the protein helix. Both the stretch modulus and the bending rigidity of a fiber change in the presence of various nucleotide cofactors-e.g., [gamma-thio]ATP, ATP, and ADP-indicating a substantial re-arrangement of spatial relationships between the nucleic acid and the protein scaffold. In particular, when complexed with ATP, a fiber becomes twice as extensible as a [gamma-thio]ATP fiber, suggesting that 32% of the DNA-binding sites have been released in its core. Such release may enable easy rotation of the DNA within the protein helix or slippage of the DNA through the center of the protein helix. PMID- 10468571 TI - Compactness of the denatured state of a fast-folding protein measured by submillisecond small-angle x-ray scattering. AB - Time-resolved small-angle x-ray scattering was used to measure the radius of gyration of cytochrome c after initiation of folding by a pH jump. Submillisecond time resolution was obtained with a microfabricated diffusional mixer and synchrotron radiation. The results show that the protein first collapses to compact denatured structures before folding very fast to the native state. PMID- 10468572 TI - The propagation of binding interactions to remote sites in proteins: analysis of the binding of the monoclonal antibody D1.3 to lysozyme. AB - The interaction of a ligand with a protein occurs at a local site (the binding site) and involves only a few residues; however, the effects of that interaction are often propagated to remote locations. The chain of events initiated by binding provides the basis for fundamental biological phenomena such as allosterism, signal transduction, and structural-stability modification. In this paper, a structure-based statistical thermodynamic approach is presented and used to predict the propagation of the stabilization effects triggered by the binding of the monoclonal antibody D1.3 to hen egg white lysozyme. Previously, Williams et al. [Williams, D. C., Benjamin, D. C., Poljak, R. J. & Rule, G. S. (1996) J. Mol. Biol. 257, 866-876] showed that the binding of this antibody affects the stability of hen egg white lysozyme and that the binding effects propagate to a selected number of residues at remote locations from the binding epitope. In this paper, we show that this phenomenon can be predicted from structure. The formalism presented here permits the identification of the structural path followed by cooperative interactions that originate at the binding site. It is shown that an important condition for the propagation of binding effects to distal regions is the presence of a significant fraction of residues with low structural stability in the uncomplexed binding site. A survey of protein structures indicates that many binding sites have a dual character and are defined by regions of high and low structural stabilities. The low-stability regions might be involved in the transmission of binding information to other regions in the protein. PMID- 10468573 TI - Rapid characterization of green fluorescent protein fusion proteins on the molecular and cellular level by fluorescence correlation microscopy. AB - Fluorescence correlation microscopy (FCM) was applied to characterize fusion proteins of the green fluorescent protein (GFP) on the cellular as well as molecular level within seconds in an integrated instrument. FCM combines the inherent sensitivity and high spatial resolution of fluorescence correlation spectroscopy with fluorescence imaging and micropositioning, thereby providing a spectrum of molecular information in the cellular context. Signatures of characteristic parameters derived from the autocorrelation functions served to distinguish a GFP fusion protein of the epidermal growth factor receptor from GFP fluorescence in the endoplasmic reticulum and cytoplasm. Diffusion constants measured for free transiently expressed GFP reproduced values reported previously with other techniques. The accessible concentration range extends from millions to only a few thousand molecules per cell, with single molecule detectability in the femtoliter detection volume. The detailed molecular characterization offered by FCM is fully compatible with automation in sample identification and detection, offering new possibilities for highly integrated high-throughput screening. PMID- 10468574 TI - Determinants of aminoglycoside-binding specificity for rRNA by using mass spectrometry. AB - We have developed methods for studying the interactions between small molecules and RNA and have applied them to characterize the binding of three classes of aminoglycoside antibiotics to ribosomal RNA subdomains. High-resolution MS was used to quantitatively identify the noncovalent binding interactions between mixtures of aminoglycosides and multiple RNA targets simultaneously. Signal overlap among RNA targets was avoided by the addition of neutral mass tags that direct each RNA target to a unique region of the spectrum. In addition to determining binding affinities, the locations of the binding sites on the RNAs were identified from a protection pattern generated by fragmenting the aminoglycoside/RNA complex. Specific complexes were observed for the prokaryotic rRNA A-site subdomain with ribostamycin, paromomycin, and lividomycin, whereas apramycin preferentially formed a complex with the eukaryotic subdomain. We show that differences in binding between paromomycin and ribostamycin can be probed by using an MS-MS protection assay. We have introduced specific base substitutions in the RNA models and have measured their impact on binding affinity and selectivity. The binding of apramycin to the prokaryotic subdomain strongly depends on the identity of position 1408, as evidenced by the selective increase in affinity for an A1408G mutant. An A1409-G1491 mismatch pair in the prokaryotic subdomain enhanced the binding of tobramycin and bekanamycin. These observations demonstrate the power of MS-based methods to provide molecular insights into small molecule/RNA interactions useful in the design of selective new antimicrobial drugs. PMID- 10468575 TI - Rotational symmetry of the C ring and a mechanism for the flagellar rotary motor. AB - FliG, FliM, and FliN, key proteins for torque generation, are located in two rings. The first protein is in the M ring and the last two are in the C ring. The rotational symmetries of the C and M rings have been determined to be about 34 (this paper) and 26 (previous work), respectively. The mechanism proposed here depends on the symmetry mismatch between the rings: the C ring extends 34 levers, of which 26 can bind to the 26 equivalent sites on the M ring. The remaining 8 levers bind to proton-pore complexes (studs) to form 8 torque generators. Movement results from the swapping of stud-bound levers with M ring-bound levers. The model predicts that both the M and C rings rotate in the same direction but at different speeds. PMID- 10468576 TI - Pearling in cells: a clue to understanding cell shape. AB - Gradual disruption of the actin cytoskeleton induces a series of structural shape changes in cells leading to a transformation of cylindrical cell extensions into a periodic chain of "pearls." Quantitative measurements of the pearling instability give a square-root behavior for the wavelength as a function of drug concentration. We present a theory that explains these observations in terms of the interplay between rigidity of the submembranous actin shell and tension that is induced by boundary conditions set by adhesion points. The theory allows estimation of the rigidity and thickness of this supporting shell. The same theoretical considerations explain the shape of nonadherent edges in the general case of untreated cells. PMID- 10468577 TI - A tubular endosomal fraction from rat liver: biochemical evidence of receptor sorting by default. AB - We previously have isolated an endosomal fraction from rat liver, termed receptor recycling compartment (RRC), which is highly enriched in recycling receptors and in the transcytotic polymeric Ig receptor (pIgR). We now have analyzed the RRC fraction by immunoisolation and found that no uniquely transcytotic elements were present, because recycling receptors and the pIgR were coisolated on the same elements. In addition, RRC was very rich in proteins previously shown to be associated with recycling endosomes, such as rab 11, cellubrevin, and endobrevin, but relatively poor in early endosome antigen 1. As RRC contains mainly tubules and small vesicles, our results indicate that it is enriched in elements of a tubular endosomal compartment involved in receptor sorting. Biochemical analysis showed that the density of recycling receptors and transcytotic pIgR in RRC membranes was similar to that in early endosome membranes. This observation supports the idea that increasing membrane surface area by endosome tubulation is the main mechanism to ensure efficient receptor sorting and, at the same time, locates RRC in a common step of the endocytotic system before final receptor segregation into distinct recycling and transcytotic pathways. PMID- 10468578 TI - Impaired glucose homeostasis and neonatal mortality in hepatocyte nuclear factor 3alpha-deficient mice. AB - Hepatocyte nuclear factors 3 (HNF-3) belong to an evolutionarily conserved family of transcription factors that are critical for diverse biological processes such as development, differentiation, and metabolism. To study the physiological role of HNF-3alpha, we generated mice that lack HNF-3alpha by homologous recombination in embryonic stem cells. Mice homozygous for a null mutation in the HNF-3alpha gene develop a complex phenotype that is characterized by abnormal feeding behavior, progressive starvation, persistent hypoglycemia, hypotriglyceridemia, wasting, and neonatal mortality between days 2 and 14. Hypoglycemia in HNF-3alpha null mice leads to physiological counter-regulatory responses in glucocorticoid and growth hormone production and an inhibition of insulin secretion but fails to stimulate glucagon secretion. Glucagon-producing pancreatic alpha cells develop normally in HNF-3alpha-/- mice, but proglucagon mRNA levels are reduced 50%. Furthermore, the transcriptional levels of neuropeptide Y are also significantly reduced shortly after birth, implying a direct role of HNF-3alpha in the expression of these genes. In contrast, mRNA levels were increased in HNF-3 target genes phosphofructo-2-kinase/fructose-2,6-bisphophatase, insulin growth factor binding protein-1, and hexokinase I of HNF-3alpha-null mice. Mice lacking one or both HNF-3alpha alleles also show impaired insulin secretion and glucose intolerance after an intraperitoneal glucose challenge, indicating that pancreatic beta-cell function is also compromised. Our results indicate that HNF 3alpha plays a critical role in the regulation of glucose homeostasis and in pancreatic islet function. PMID- 10468579 TI - Cubilin, the endocytic receptor for intrinsic factor-vitamin B(12) complex, mediates high-density lipoprotein holoparticle endocytosis. AB - Receptors that endocytose high-density lipoproteins (HDL) have been elusive. Here yolk-sac endoderm-like cells were used to identify an endocytic receptor for HDL. The receptor was isolated by HDL affinity chromatography and identified as cubilin, the recently described endocytic receptor for intrinsic factor-vitamin B(12). Cubilin antibodies inhibit HDL endocytosis by the endoderm-like cells and in mouse embryo yolk-sac endoderm, a prominent site of cubilin expression. Cubilin-mediated HDL endocytosis is inhibitable by HDL(2), HDL(3), apolipoprotein (apo)A-I, apoA-II, apoE, and RAP, but not by low-density lipoprotein (LDL), oxidized LDL, VLDL, apoC-I, apoC-III, or heparin. These findings, coupled with the fact that cubilin is expressed in kidney proximal tubules, suggest a role for this receptor in embryonic acquisition of maternal HDL and renal catabolism of filterable forms of HDL. PMID- 10468580 TI - Syntaxin 1 interacts with the L(D) subtype of voltage-gated Ca(2+) channels in pancreatic beta cells. AB - Interaction of syntaxin 1 with the alpha(1D) subunit of the voltage-gated L type Ca(2+) channel was investigated in the pancreatic beta cell. Coexpression of the enhanced green fluorescent protein-linked alpha(1D) subunit with the enhanced blue fluorescent protein-linked syntaxin 1 and Western blot analysis together with subcellular fractionation demonstrated that the alpha(1D) subunit and syntaxin 1 were colocalized in the plasma membrane. Furthermore, the alpha(1D) subunit was coimmunoprecipitated efficiently by a polyclonal antibody against syntaxin 1. Syntaxin 1 also played a central role in the modulation of L type Ca(2+) channel activity because there was a faster Ca(2+) current run-down in cells incubated with antisyntaxin 1 compared with controls. In parallel, antisyntaxin 1 markedly reduced insulin release in both intact and permeabilized cells, subsequent to depolarization with K(+) or exposure to high Ca(2+). Exchanging Ca(2+) for Ba(2+) abolished the effect of antisyntaxin 1 on both Ca(2+) channel activity and insulin exocytosis. Moreover, antisyntaxin 1 had no significant effects on Ca(2+)-independent insulin release trigged by hypertonic stimulation. This suggests that there is a structure-function relationship between the alpha(1D) subunit of the L type Ca(2+) channel and the exocytotic machinery in the pancreatic beta cell. PMID- 10468581 TI - Role for yeast inhibitor of apoptosis (IAP)-like proteins in cell division. AB - Inhibitors of apoptosis (IAPs) are a family of proteins that bear baculoviral IAP repeats (BIRs) and regulate apoptosis in vertebrates and Drosophila melanogaster. The yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe both encode a single IAP, designated BIR1 and bir1, respectively, each of which bears two BIRs. In rich medium, BIR1 mutant S. cerevisiae underwent normal vegetative growth and mitosis. Under starvation conditions, however, BIR1 mutant diploids formed spores inefficiently, instead undergoing pseudohyphal differentiation. Most spores that did form failed to survive beyond two divisions after germination. bir1 mutant S. pombe spores also died in the early divisions after spore germination and became blocked at the metaphase/anaphase transition because of an inability to elongate their mitotic spindle. Rather than inhibiting caspase-mediated cell death, yeast IAP proteins have roles in cell division and appear to act in a similar way to the IAPs from Caenorhabditis elegans and the mammalian IAP Survivin. PMID- 10468582 TI - An invasion-associated Salmonella protein modulates the actin-bundling activity of plastin. AB - The entry of Salmonella typhimurium into nonphagocytic cells requires a panel of bacterial effector proteins that are delivered to the host cell via a type III secretion system. These proteins modulate host-cell signal-transduction pathways and the actin cytoskeleton to induce membrane ruffling and bacterial internalization. One of these bacterial effectors, termed SipA, is an actin binding protein that is required for efficient Salmonella entry into host cells. We report here that SipA forms a complex with T-plastin on bacterial infection. Formation of such a complex, which requires the presence of F-actin, results in a marked increase in the actin-bundling activity of T-plastin. We also report that T-plastin is recruited to S. typhimurium-induced membrane ruffles by a CDC42 dependent signaling process and is required for bacterial entry. We propose that modulation of the actin-bundling activity of T-plastin by SipA results in the stabilization of the actin filaments at the point of bacterial-host cell contact, which leads to more efficient Salmonella internalization. PMID- 10468583 TI - The tumor-suppressor activity of PTEN is regulated by its carboxyl-terminal region. AB - PTEN is a recently identified tumor suppressor inactivated in a variety of cancers such as glioblastoma and endometrial and prostate carcinoma. It contains an amino-terminal phosphatase domain and acts as a phosphatidylinositol 3,4,5 trisphosphate phosphatase antagonizing the activity of the phosphatidylinositol 3 OH kinase. PTEN also contains a carboxyl-terminal domain, and we addressed the role of this region that, analogous to the amino-terminal phosphatase domain, is the target of many mutations identified in tumors. Expression of carboxyl terminal mutants in PTEN-deficient glioblastoma cells permitted the anchorage independent growth of the cells that otherwise was suppressed by wild-type PTEN. The stability of these mutants in cells was reduced because of rapid degradation. Although the carboxyl-terminal region contains regulatory PEST sequences and a PDZ-binding motif, these specific elements were dispensable for the tumor suppressor function. The study of carboxyl-terminal point mutations affecting the stability of PTEN revealed that these were located in strongly predicted beta strands. Surprisingly, the phosphatase activity of these mutants was affected in correlation with the degree of disruption of these structural elements. We conclude that the carboxyl-terminal region is essential for regulating PTEN stability and enzymatic activity and that mutations in this region are responsible for the reversion of the tumor-suppressor phenotype. We also propose that the molecular conformational changes induced by these mutations constitute the mechanism for PTEN inactivation. PMID- 10468584 TI - A nuclear protein tyrosine phosphatase is required for the inactivation of Stat1. AB - The Stat1 activation-inactivation cycle involves phosphorylation of Stat1 in the cytoplasm, translocation to the nucleus, and then a return of the protein to the cytoplasm in a dephosphorylated state. However, the intracellular site of Stat1 dephosphorylation has not been determined. As receptor signaling declines, the flow of activated Stat1 molecules should be to the site of their dephosphorylation. We found that upon receptor-Janus kinase inactivation, either gradual or abruptly induced by staurosporine treatment, the flow of Stat1 was from cytoplasm to the nucleus and the nucleus was the final compartment in which phosphorylated Stat1 was detected. N-terminal mutants of Stat1, previously shown to remain phosphorylated for a longer time than wild-type Stat1, were able to enter the nucleus and were not inactivated in the presence of staurosporine, directly demonstrating that these mutations affect phosphatase access and/or activity during the normal dephosphorylation of Stat1. In the presence of sodium vanadate, a phosphatase inhibitor, phosphorylated Stat1 accumulated in the nucleus as the total amount of Stat1 in the cytoplasm declined to low levels. We conclude that the nucleus is the site of Stat1 inactivation and that dephosphorylation is required for the rapid nuclear export of Stat1. PMID- 10468585 TI - The hsp70-associating protein Hap46 binds to DNA and stimulates transcription. AB - We investigated the ubiquitously expressed hsp70-associating protein Hap46, which is also called RAP46 and is homologous to BAG-1, for activities independent of hsp70 interactions. We observed in vitro binding to various DNA fragments but detected no apparent sequence specificity. Deletion of the amino-terminal decapeptide, which contains two clusters of three basic amino acids each, abolished the DNA-binding ability of Hap46. Similarly, exchange of either of these positively charged clusters for three alanines resulted in loss of DNA binding. Using a fusion of Hap46 and green fluorescent protein, we found preferential accumulation in cell nuclei on heat stress as compared with unstressed cells. The repressive effect of heat shock on overall transcriptional activity in human DU145 carcinoma cells was largely prevented when Hap46 was overexpressed by transfection. Such overproduction of Hap46 also resulted in enhanced expression of specific reporter gene constructs and in increased levels of mRNAs specific for hsp70 and hsp40 after temperature stress. In vitro transcription with nuclear extracts was stimulated greatly by Hap46. Like DNA binding, transcriptional enhancement required amino-terminally located basic amino acid residues but not the carboxyl-terminal portion of Hap46 known to participate in hsp70 interaction. Our results show that Hap46 is a bifunctional protein that can interact with both hsp70s and DNA, employing different portions of the molecule. They also suggest that Hap46 is involved in temperature sensitive regulation of transcription, acting as a general transcriptional activator. PMID- 10468586 TI - Altered dosage of the Saccharomyces cerevisiae spindle pole body duplication gene, NDC1, leads to aneuploidy and polyploidy. AB - Saccharomyces cerevisiae cells are exquisitely sensitive to altered dosage of the spindle pole body duplication gene, NDC1. We show that the NDC1 locus is haploinsufficient because diploid yeast cells cannot survive with a single chromosomal copy of the NDC1 gene. Diploid cells with a single copy of NDC1 can survive by gaining an extra copy of the NDC1-containing chromosome. NDC1 haploinsufficiency is a dominant loss-of-function phenotype that leads to aneuploidy. Furthermore, we report that overexpression of NDC1 leads to spindle pole body duplication defects indistinguishable from those observed in ndc1-1 mutant cells. Cells overexpressing NDC1 arrest with monopolar spindles and exhibit increase-in-ploidy phenotypes. Thus, both increased and decreased NDC1 dosage can lead to aneuploidy. The striking sensitivity of yeast cells to changes in NDC1 gene dosage suggests a model for the behavior of some tumor suppressor genes and oncogenes in which loss-of-function mutations and overexpression, respectively, lead to increased genetic instability. PMID- 10468587 TI - Evidence for a conserved system for iron metabolism in the mitochondria of Saccharomyces cerevisiae. AB - nifU of nitrogen-fixing bacteria is involved in the synthesis of the Fe-S cluster of nitrogenase. In a synthetic lethal screen with the mitochondrial heat shock protein (HSP)70, SSQ1, we identified a gene of Saccharomyces cerevisiae, NFU1, which encodes a protein with sequence identity to the C-terminal domain of NifU. Two other yeast genes were found to encode proteins related to the N-terminal domain of bacterial NifU. They have been designated ISU1 and ISU2. Isu1, Isu2, and Nfu1 are located in the mitochondrial matrix. ISU genes of yeast carry out an essential function, because a Deltaisu1Deltaisu2 strain is inviable. Growth of Deltanfu1Delta isu1 cells is significantly compromised, allowing assessment of the physiological roles of Nfu and Isu proteins. Mitochondria from Deltanfu1Deltaisu1 cells have decreased activity of several respiratory enzymes that contain Fe-S clusters. As a result, Deltanfu1Deltaisu1 cells grow poorly on carbon sources requiring respiration. Deltanfu1Deltaisu1 cells also accumulate abnormally high levels of iron in their mitochondria, similar to Deltassq1 cells, indicating a role for these proteins in iron metabolism. We suggest that NFU1 and ISU1 gene products play a role in iron homeostasis, perhaps in assembly, insertion, and/or repair of mitochondrial Fe-S clusters. The conservation of these protein domains in many organisms suggests that this role has been conserved throughout evolution. PMID- 10468589 TI - Lepidopteran DALP, and its mammalian ortholog HIC-5, function as negative regulators of muscle differentiation. AB - During myogenesis, reductions in trophic factor availability signal most myoblasts to fuse, up-regulate the expression of muscle-specific genes, and form myotubes. Those cells failing to differentiate into myotubes initiate apoptosis and rapidly die. At present, the signal-transduction molecules that determine whether myoblasts should differentiate or die are largely unknown. In this report, we describe the cloning and characterization of DALP, a small LIM-only type zinc-finger protein that is induced when the intersegmental muscles (ISMs) of the moth Manduca sexta become committed to die at the end of metamorphosis. Forced expression of death-associated LIM-only protein (DALP) in Drosophila results in skeletal muscle atrophy. Ectopic expression of DALP, or its mammalian ortholog Hic-5, blocks differentiation and induces apoptosis in mouse C(2)C(12) myoblasts. Both of these effects can be overcome by contact with normal myoblasts or by ectopic expression of the muscle-specific transcription factor MyoD. Hic-5 expression is specifically and dramatically induced in normal myoblasts that die after removal of trophic support. Taken together, these data suggest that DALP and Hic-5 act upstream of MyoD and function as phylogenetically conserved "switches" to block muscle differentiation and induce death. PMID- 10468590 TI - Homeotic genes and the arthropod head: expression patterns of the labial, proboscipedia, and Deformed genes in crustaceans and insects. AB - cDNA fragments of the homologues of the Drosophila head homeotic genes labial (lab), proboscipedia (pb), and Deformed (Dfd) have been isolated from the crustacean Porcellio scaber. Because the accumulation domains of the head homeotic complex (Hox) genes had not been previously reported for crustaceans, we studied the expression patterns of these genes in P. scaber embryos by using in situ hybridization. The P. scaber lab homologue is expressed in the developing second antennal segment and its appendages. This expression domain in crustaceans and in the homologous intercalary segment of insects suggests that the lab gene specified this metamere in the last common ancestor of these two groups. The expression domain of the P. scaber pb gene is in the posterior part of the second antennal segment. This domain, in contrast to that in insects, is colinear with the domains of other head genes in P. scaber, and it differs from the insect pb gene expression domain in the posterior mouthparts, suggesting that the insect and crustacean patterns evolved independently from a broader ancestral domain similar to that found in modern chelicerates. P. scaber Dfd is expressed in the mandibular segment and paragnaths (a pair of ventral mouthpart structures associated with the stomodeum) and differs from insects, where expression is in the mandibular and maxillary segments. Thus, like pb, Dfd shows a divergent Hox gene deployment. We conclude that homologous structures of the mandibulate head display striking differences in their underlying developmental programs related to Hox gene expression. PMID- 10468591 TI - Replicated evolution of trophic specializations in an endemic cichlid fish lineage from Lake Tanganyika. AB - The current phylogenetic hypothesis for the endemic Lake Tanganyika cichlid fishes of the tribe Eretmodini is based solely on morphology and suggests that more complex trophic morphologies derived only once from a less specialized ancestral condition. A molecular phylogeny of eretmodine cichlids based on partial mitochondrial DNA cytochrome b and control-region sequences was used to reconstruct the evolutionary sequence of trophic adaptations and to test alternative models of morphological divergence. The six mitochondrial lineages found disagree with the current taxonomy and the morphology-based phylogeny. Mitochondrial lineages with similar trophic morphologies are not grouped monophyletically but are typically more closely related to lineages with different trophic phenotypes currently assigned to other genera. Our results indicate multiple independent origins of similar trophic specializations in these cichlids. A pattern of repeated divergent morphological evolution becomes apparent when the phylogeography of the mitochondrial haplotypes is analyzed in the context of the geological and paleoclimatological history of Lake Tanganyika. In more than one instance within Lake Tanganyika, similar morphological divergence of dentitional traits occurred in sympatric species pairs. Possibly, resource-based divergent selective regimes led to resource partitioning and brought about similar trophic morphologies independently and repeatedly. PMID- 10468588 TI - Transcription repression by Xenopus ET and its human ortholog TBX3, a gene involved in ulnar-mammary syndrome. AB - T box (Tbx) genes are a family of developmental regulators with more than 20 members recently identified in invertebrates and vertebrates. Mutations in Tbx genes have been found to cause several human diseases. Our understanding of functional mechanisms of Tbx products has come mainly from the prototypical T/Brachyury, which is a transcription activator. We previously discovered ET, a Tbx gene expressed in Xenopus embryos. We report here that ET is an ortholog of the human Tbx3 and that ET is a repressor of basal and activated transcription. Functional dissection of the ET protein reveals a novel transcription-repression domain highly conserved among ET, human TBX3, and TBX2. These results reveal a new transcription repressor domain, show the existence of a subfamily of transcription repressors in the Tbx superfamily, and provide a basis for understanding etiology of diseases caused by Tbx3 mutations. PMID- 10468592 TI - Genetic benefits enhance the reproductive success of polyandrous females. AB - Although it is generally accepted that females can gain material benefits by mating with more than one male, the proposal that polyandry provides genetic benefits remains controversial, largely because direct experimental support is lacking. Here, we report the results of a study testing for genetic benefits to polyandry in the pseudoscorpion Cordylochernes scorpioides. In an experiment that controlled for male mating experience and the number of spermatophores accepted by a female, twice-mated females received either one sperm-packet from each of two different males (the "DM" treatment) or two sperm-packets from a single male (the same male or "SM" treatment). Over their lifetime, DM females gave birth to 32% more offspring than did SM females, primarily because of a significantly reduced rate of spontaneous abortion. This result could not be attributed to male infertility nor to lack of sexual receptivity in males paired with previous mates. Spermatophore and sperm numbers did not differ between males presented with a previous mate and males paired with a new female. Because SM and DM females received the same quantity of ejaculate, it was possible to eliminate material benefits as a contributor to the enhanced reproductive success of DM females. The reduction in embryo failure rate achieved by DM females is most consistent with the genetic incompatibility avoidance hypothesis, i.e., that polyandry enables females to exploit postcopulatory mechanisms for reducing the risk and/or cost of fertilization by genetically incompatible sperm. This study, which rigorously controlled for material benefits and excluded inbreeding effects, demonstrates that polyandry provides genetic benefits that significantly enhance female lifetime reproductive success. PMID- 10468593 TI - Evolutionary consequences of niche construction and their implications for ecology. AB - Organisms regularly modify local resource distributions, influencing both their ecosystems and the evolution of traits whose fitness depends on such alterable sources of natural selection in environments. We call these processes niche construction. We explore the evolutionary consequences of niche construction using a two-locus population genetic model, which extends earlier analyses by allowing resource distributions to be influenced both by niche construction and by independent processes of renewal and depletion. The analysis confirms that niche construction can be a potent evolutionary agent by generating selection that leads to the fixation of otherwise deleterious alleles, supporting stable polymorphisms where none are expected, eliminating what would otherwise be stable polymorphisms, and generating unusual evolutionary dynamics. Even small amounts of niche construction, or niche construction that only weakly affects resource dynamics, can significantly alter both ecological and evolutionary patterns. PMID- 10468594 TI - The complete chloroplast DNA sequence of the green alga Nephroselmis olivacea: insights into the architecture of ancestral chloroplast genomes. AB - Green plants seem to form two sister lineages: Chlorophyta, comprising the green algal classes Prasinophyceae, Ulvophyceae, Trebouxiophyceae, and Chlorophyceae, and Streptophyta, comprising the Charophyceae and land plants. We have determined the complete chloroplast DNA (cpDNA) sequence (200,799 bp) of Nephroselmis olivacea, a member of the class (Prasinophyceae) thought to include descendants of the earliest-diverging green algae. The 127 genes identified in this genome represent the largest gene repertoire among the green algal and land plant cpDNAs completely sequenced to date. Of the Nephroselmis genes, 2 (ycf81 and ftsI, a gene involved in peptidoglycan synthesis) have not been identified in any previously investigated cpDNA; 5 genes [ftsW, rnE, ycf62, rnpB, and trnS(cga)] have been found only in cpDNAs of nongreen algae; and 10 others (ndh genes) have been described only in land plant cpDNAs. Nephroselmis and land plant cpDNAs share the same quadripartite structure-which is characterized by the presence of a large rRNA-encoding inverted repeat and two unequal single-copy regions-and very similar sets of genes in corresponding genomic regions. Given that our phylogenetic analyses place Nephroselmis within the Chlorophyta, these structural characteristics were most likely present in the cpDNA of the common ancestor of chlorophytes and streptophytes. Comparative analyses of chloroplast genomes indicate that the typical quadripartite architecture and gene-partitioning pattern of land plant cpDNAs are ancient features that may have been derived from the genome of the cyanobacterial progenitor of chloroplasts. Our phylogenetic data also offer insight into the chlorophyte ancestor of euglenophyte chloroplasts. PMID- 10468595 TI - Constructing primate phylogenies from ancient retrovirus sequences. AB - The genomes of modern humans are riddled with thousands of endogenous retroviruses (HERVs), the proviral remnants of ancient viral infections of the primate lineage. Most HERVs are nonfunctional, selectively neutral loci. This fact, coupled with their sheer abundance in primate genomes, makes HERVs ideal for exploitation as phylogenetic markers. Endogenous retroviruses (ERVs) provide phylogenetic information in two ways: (i) by comparison of integration site polymorphism and (ii) by orthologous comparison of evolving, proviral, nucleotide sequence. In this study, trees are constructed with the noncoding long terminal repeats (LTRs) of several ERV loci. Because the two LTRs of an ERV are identical at the time of integration but evolve independently, each ERV locus can provide two estimates of species phylogeny based on molecular evolution of the same ancestral sequence. Moreover, tree topology is highly sensitive to conversion events, allowing for easy detection of sequences involved in recombination as well as correction for such events. Although other animal species are rich in ERV sequences, the specific use of HERVs in this study allows comparison of trees to a well established phylogenetic standard, that of the Old World primates. HERVs, and by extension the ERVs of other species, constitute a unique and plentiful resource for studying the evolutionary history of the Retroviridae and their animal hosts. PMID- 10468596 TI - Phylogenetic relationships among cetartiodactyls based on insertions of short and long interpersed elements: hippopotamuses are the closest extant relatives of whales. AB - Insertion analysis of short and long interspersed elements is a powerful method for phylogenetic inference. In a previous study of short interspersed element data, it was found that cetaceans, hippopotamuses, and ruminants form a monophyletic group. To further resolve the relationships among these taxa, we now have isolated and characterized 10 additional loci. A phylogenetic analysis of these data was able to resolve relationships among the major cetartiodactyl groups, thereby shedding light on the origin of whales. The results indicated (i) that cetaceans are deeply nested within Artiodactyla, (ii) that cetaceans and hippopotamuses form a monophyletic group, (iii) that pigs and peccaries form a monophyletic group to the exclusion of hippopotamuses, (iv) that chevrotains diverged first among ruminants, and (v) that camels diverged first among cetartiodactyls. These findings lead us to conclude that cetaceans evolved from an immediate artiodactyl, not mesonychian, ancestor. PMID- 10468597 TI - Late changes in spliceosomal introns define clades in vertebrate evolution. AB - The evolutionary origin of spliceosomal introns has been the subject of much controversy. Introns are proposed to have been both lost and gained during evolution. If the gain or loss of introns are unique events in evolution, they can serve as markers for phylogenetic analysis. We have made an extensive survey of the phylogenetic distribution of seven spliceosomal introns that are present in Fugu genes, but not in their mammalian homologues; we show that these introns were acquired by actinopterygian (ray-finned) fishes at various stages of evolution. We have also investigated the intron pattern of the rhodopsin gene in fishes, and show that the four introns found in the ancestral chordate rhodopsin gene were simultaneously lost in a common ancestor of ray-finned fishes. These changes in introns serve as excellent markers for phylogenetic analysis because they reliably define clades. Our intron-based cladogram establishes the difficult to-ascertain phylogenetic relationships of some ray-finned fishes. For example, it shows that bichirs (Polypterus) are the sister group of all other extant ray finned fishes. PMID- 10468598 TI - Developmental shifts and species selection in gastropods. AB - The fossil record of marine gastropods has been used as evidence to support the operation of species selection; namely, that species with limited dispersal differentially increase in numbers because they are more likely to speciate than widely dispersing species. This conclusion is based on a tacit phylogenetic assumption that increases in species with limited dispersal are solely the result of speciation within monophyletic groups with low dispersal. To test this assumption, we reconstructed a phylogeny from nuclear sequence data for 70 species of the marine gastropod genus Conus and used it to map the evolution of developmental mode. All eight species without planktonic life history phases recently and independently evolved this characteristic from ancestors with planktonic larval phases, showing that transitions in developmental mode are common in this group. A simple model of species diversification shows that such shifts can control the relative numbers of species with and without dispersing larval stages, leading to apparent species selection. Such results challenge the conclusion that increases in the number of nonplanktonic species relative to species with planktonic larvae over geologic time is necessarily a result of higher rates of speciation of nonplanktonic lineages and show that demonstration of species selection requires a phylogenetic framework. PMID- 10468599 TI - Three new allelic mouse mutations that cause skeletal overgrowth involve the natriuretic peptide receptor C gene (Npr3). AB - In 1979, a BALB/cJ mouse was identified with an exceptionally long body. This phenotype was found to be caused by a recessive mutation, designated longjohn (lgj), that mapped to the proximal region of chromosome 15. Several years later, a mouse with a similarly elongated body was identified in an outbred stock after chemical mutagenesis with ethylnitrosourea. This phenotype also was caused by a recessive mutation, designated strigosus (stri). The two mutations were found to be allelic. A third allele was identified in a DBA/2J mouse and was designated longjohn-2J (lgj(2J)). Analysis of skeletal preparations of stri/stri mice indicated that the endochondral ossification process was slightly delayed, resulting in an extended proliferation zone. A recent study reported that mice overexpressing brain natriuretic peptide, one of the members of the natriuretic peptide family, exhibit a skeletal-overgrowth syndrome with endochondral ossification defects. The Npr3 gene coding for type C receptor for natriuretic peptides (NPR-C), which is mainly involved in the clearance of the natriuretic peptides, mapped in the vicinity of our mouse mutations and thus was a candidate gene. The present study reports that all three mutations involve the Npr3 gene and provides evidence in vivo that there is a natriuretic-related bone pathway, underscoring the importance of natriuretic peptide clearance by natriuretic peptide type C receptor. PMID- 10468600 TI - Rice gibberellin-insensitive dwarf mutant gene Dwarf 1 encodes the alpha-subunit of GTP-binding protein. AB - A rice Dwarf 1 gene was identified by using a map-based cloning strategy. Its recessive mutant allele confers a dwarf phenotype. Linkage analysis revealed that a cDNA encoding the alpha-subunit of GTP-binding protein cosegregated with d1 in 3,185 d1 segregants. Southern hybridization analysis with this cDNA as a probe showed different band patterns in several d1 mutant lines. In at least four independent d1 mutants, no gene transcript was observed by Northern hybridization analysis. Sequencing analysis revealed that an 833-bp deletion had occurred in one of the mutant alleles, which resulted in an inability to express GTP-binding protein. A transgenic d1 mutant with GTP-binding protein gene restored the normal phenotype. We conclude that the rice Dwarf 1 gene encodes GTP-binding protein and that the protein plays an important role in plant growth and development. Because the d1 mutant is classified as gibberellin-insensitive, we suggest that the GTP binding protein might be associated with gibberellin signal transduction. PMID- 10468601 TI - An enhancer element located downstream of the major glutamate dehydrogenase gene of Bacillus subtilis. AB - The rocG gene of Bacillus subtilis, encoding a catabolic glutamate dehydrogenase, is transcribed by SigL (sigma(54))-containing RNA polymerase and requires for its expression RocR, a member of the NtrC/NifA family of proteins that bind to enhancer-like elements, called upstream activating sequences (UAS). Unlike the case for other sigma(54)-dependent genes, rocG has no UAS; instead, its expression depends on a sequence located 1.5 kilobases downstream of the rocG promoter, beyond the end of the rocG coding region. The same sequence also serves as the UAS for the downstream rocABC operon and can activate rocG if moved upstream of its promoter. Furthermore, the activating sequence can be moved as far as 15 kilobases downstream of the rocG promoter and still retain partial activity. PMID- 10468602 TI - Mechanisms of inactivation of mismatch repair genes in human colorectal cancer cell lines: the predominant role of hMLH1. AB - Fifteen to twenty-five percent of sporadic colorectal carcinomas are replication error (RER) positive. Because the frequency of mutations in the mismatch repair genes (hMLH1 and hMSH2) is low in these tumors, we have investigated the role of mutational inactivation, methylation of the promoter region, and loss of heterozygosity (LOH) as a possible explanation for the mutator phenotype of RER+ colorectal cancer cell lines. Genomic DNA was extracted from a panel of 49 human colorectal cancer cell lines. The RER status was determined by amplification of BAT-26. All exons of hMLH1 and hMSH2 were amplified with the PCR and screened by using single-strand conformational polymorphism and direct sequencing. The methylation status was ascertained by methylation-specific PCR after bisulfite modification of DNA. Western blotting for hMLH1 was performed on methylated cell lines before and after the addition of the demethylating agent 5-azacytidine. LOH was sought by GENESCAN analysis of amplified CA repeat markers and indirectly by determining the number of homozygotes in the cell lines and human random controls. Twelve cell lines from ten tumors (24%) were RER+. Hypermethylation of the hMLH1 promoter occurred in five of ten (50%) RER+ tumors, whereas three of thirty-two (6%) RER tumors showed partial methylation. None of the fully methylated cell lines expressed hMLH1, although all reexpressed hMLH1 after treatment with 5-azacytidine. There was no LOH in the RER+ tumors in either hMLH1 or hMSH2. Our results suggest that mutations of hMLH1 together with hypermethylation of the promoter region, but not LOH, are the cause of the mutator phenotype in the majority (70%) of RER+ tumors. PMID- 10468603 TI - Rare germinal unequal crossing-over leading to recombinant gene formation and gene duplication in Arabidopsis thaliana. AB - Small, multigene families organized in a tandem array can facilitate the rapid evolution of the gene cluster by a process of meiotic unequal crossing-over. To study this process in a multicellular organism, we created a synthetic RBCSB gene cluster in Arabidopsis thaliana and used this to measure directly the frequency of meiotic, intergenic unequal crossing-over between sister chromatids. The synthetic RBCSB gene cluster was composed of a silent DeltaRBCS1B::LUC chimeric gene fusion, lacking all 5' transcription and translation signals, followed by RBCS2B and RBC3B genomic DNA. Expression of luciferase activity (luc(+)) required a homologous recombination event between the DeltaRBCS1B::LUC and the RBCS3B genes, yielding a novel recombinant RBCS3B/ 1B::LUC chimeric gene whose expression was driven by RBCS3B 5' transcription and translation signals. Using sensitive, single-photon-imaging equipment, three luc(+) seedlings were identified in more than 1 million F2 seedlings derived from self-fertilized F1 plants hemizygous for the synthetic RBCSB gene cluster. The F2 luc(+) seedlings were isolated, and molecular and genetic analysis indicated that the luc(+) trait was caused by the formation of a recombinant chimeric RBCS3B/1B::LUC gene. A predicted duplication of the RBCS2B gene also was present. The recombination resolution break points mapped adjacent to a region of intron I at which a disjunction in sequence similarity between RBCS1B and RBCS3B occurs; this provided evidence supporting models of gene cluster evolution by exon-shuffling processes. In contrast to most measures of meiotic unequal crossing-over that require the deletion of a gene in a gene cluster, these results directly measured the frequency of meiotic unequal crossing-over (approximately 3 x 10(-6)), leading to the expansion of the gene cluster and the formation of a novel recombinant gene. PMID- 10468604 TI - The HNF-4/HNF-1alpha transactivation cascade regulates gene activity and chromatin structure of the human serine protease inhibitor gene cluster at 14q32.1. AB - Hepatocyte-specific expression of the alpha1-antitrypsin (alpha1AT) gene requires the activities of two liver-enriched transactivators, hepatocyte nuclear factors 1alpha and 4 (HNF-1alpha and HNF-4). The alpha1AT gene maps to a region of human chromosome 14q32.1 that includes a related serine protease inhibitor (serpin) gene encoding corticosteroid-binding globulin (CBG), and the chromatin organization of this approximately 130-kb region, as defined by DNase I hypersensitive sites, has been described. Microcell transfer of human chromosome 14 from fibroblasts to rat hepatoma cells results in activation of alpha1AT and CBG transcription and chromatin reorganization of the entire locus. To assess the roles of HNF-1alpha and HNF-4 in gene activation and chromatin remodeling, we transferred human chromosome 14 from fibroblasts to rat hepatoma cell variants that are deficient in expression of HNF-1alpha and HNF-4. The variant cells failed to activate either alpha1AT or CBG transcription, and chromatin remodeling failed to occur. However, alpha1AT and CBG transcription could be rescued by transfecting the cells with expression plasmids encoding HNF-1alpha or HNF-4. In these transfectants, the chromatin structure of the entire alpha1AT/CBG locus was reorganized to an expressing cell-typical state. Thus, HNF-1alpha and HNF-4 control both chromatin structure and gene activity of two cell-specific genes within the serpin gene cluster at 14q32.1. PMID- 10468605 TI - Chimerism in humans after intragenic recombination at the haptoglobin locus during early embryogenesis. AB - The human haptoglobin (HP) HP*2 allele contains a 1.7-kilobase (kb) intragenic duplication that arose after a unique nonhomologous recombination between the prototype HP*1 alleles. During a genetic screening of 13,000 children of survivors exposed to atomic-bomb radiation and 10,000 children of unexposed persons, two children suspected of carrying de novo mutations at the haptoglobin locus were identified (one in each group). DNA analyses of single-cell-derived colonies of Epstein-Barr virus-transformed B cells revealed that the two children were mosaics comprising HP*2/HP*2 and HP*2/HP*1 cells at a ratio of approximately 3:1. We infer that the latter cells are caused by reversion of one HP*2 allele to HP*1 through an intramolecular homologous recombination between the duplicated segments of the Hp*2 allele that excised one of the segments. Because the mosaicism is substantial (approximately 25%), this recombination must have occurred in early embryogenesis. The frequency of finding these children and the extent of their mosaicisms corresponds to an HP*2 to HP*1 reversion rate of 8 x 10(-6) per cell during development. This leads to the prediction that the HP*1 allele also will be represented, although usually at a very low frequency, in any HP2-2 person. We tested this prediction by using PCR for a single individual and found the HP*1 allele at frequencies of 4 x 10(-6) and 3 x 10(-6) in somatic and sperm cells. The HP*1 allele was detected by PCR in all four other HP2-2 individuals, which supports the regular but rare occurrence somatically of homologous recombination within duplicated regions in humans, in agreement with previous observations in mouse and Drosophila. PMID- 10468606 TI - A physical complex of the Fanconi anemia proteins FANCG/XRCC9 and FANCA. AB - Fanconi anemia (FA) is a recessively inherited disease characterized at the cellular level by spontaneous chromosomal instability and specific hypersensitivity to cross-linking agents. FA is genetically heterogeneous, comprising at least eight complementation groups (A-H). We report that the protein encoded by the gene mutated in complementation group G (FANCG) localizes to the cytoplasm and nucleus of the cell and assembles in a molecular complex with the FANCA protein, both in vivo and in vitro. Endogenous FANCA/FANCG complex was detected in both non-FA cells and in FA cells from groups D and E. By contrast, no complex was detected in specific cell lines belonging to groups A and G, whereas reduced levels were found in cells from groups B, C, F, and H. Wild-type levels of FANCA/FANCG complex were restored upon correction of the cellular phenotype by transfection or cell fusion experiments, suggesting that this complex is of functional significance in the FA pathway. These results indicate that the cellular FA phenotype can be connected to three biochemical subtypes based on the levels of FANCA/FANCG complex. Disruption of the complex may provide an experimental strategy for chemosensitization of neoplastic cells. PMID- 10468607 TI - Recycling MHC class I molecules and endosomal peptide loading. AB - MHC class I molecules usually present peptides derived from endogenous antigens that are bound in the endoplasmic reticulum. Loading of exogenous antigens on class I molecules, e.g., in cross-priming, sometimes occurs, but the intracellular location where interaction between the antigenic fragment and class I takes place is unclear. Here we show that measles virus F protein can be presented by class I in transporters associated with antigen processing independent, NH(4)Cl-sensitive manner, suggesting that class I molecules are able to interact and bind antigen in acidic compartments, like class II molecules. Studies on intracellular transport of green fluorescent protein-tagged class I molecules in living cells confirmed that a small fraction of class I molecules indeed enters classical MHC class II compartments (MIICs) and is transported in MIICs back to the plasma membrane. Fractionation studies show that class I complexes in MIICs contain peptides. The pH in MIIC (around 5.0) is such that efficient peptide exchange can occur. We thus present evidence for a pathway for class I loading that is shared with class II molecules. PMID- 10468608 TI - Calcium signaling induces acquisition of dendritic cell characteristics in chronic myelogenous leukemia myeloid progenitor cells. AB - Effective host T lymphocyte sensitization to malignant cells depends on successful antigen presentation. In this study, we examined the capacity of malignant myeloid progenitor cells of patients in the chronic phase of chronic myelogenous leukemia (CML) to acquire characteristics of activated dendritic cells (DCs) after intracellular calcium mobilization, thereby bypassing a need for third-party antigen-presenting cells. Treatment of purified CD33(+) CML cells from 15 patients with calcium ionophore (CI) consistently resulted in de novo expression of the costimulatory molecules CD80 (B7.1) and CD86 (B7.2), CD40 and the DC-specific activation marker CD83, as well as marked up-regulation of MHC class I and II molecules and the adhesion molecule CD54. Most of these changes occurred within 24 hr of treatment. Morphologically, CI-treated CML cells developed long dendritic projections similar to those seen in mature DCs. Functionally, CI-treated CML cells provided stimulation of allogeneic T lymphocytes 10- to 20-fold that of untreated CML cells or untreated monocytes. Fluorescent in situ hybridization of CI-activated CML cells confirmed their leukemic origin by displaying the typical bcr/abl fusion signal. No difference in bcr/abl translocation percentages between untreated and CI-treated CML nuclei was observed. These observations indicate that calcium mobilization may constitute a valuable approach for rapidly and reliably generating CML-derived DCs for immunotherapy of CML. PMID- 10468609 TI - Mice lacking all conventional MHC class II genes. AB - MHC class II (MHC-II) molecules play a central role in the selection of the T cell repertoire, in the establishment and regulation of the adaptive immune response, and in autoimmune deviation. We have generated knockout mice lacking all four of the classical murine MHC-II genes (MHCII(Delta/Delta) mice), via a large (80-kilobase) deletion of the entire class II region that was engineered by homologous recombination and Cre recombinase-mediated excision. These mice feature immune system perturbations like those of Aalpha and Abeta knockout animals, notably a dearth of CD4(+) lymphocytes in the thymus and spleen. No new anatomical or physiological abnormalities were observed in MHCII(Delta/Delta) mice. Because these animals are devoid of all classical MHC-II chains, even unpaired chains, they make excellent recipients for MHC-II transgenes from other species, avoiding the problem of interspecies cross-pairing of MHC-II chains. Therefore, they should be invaluable for engineering "humanized" mouse models of human MHC-II-associated autoimmune disorders. PMID- 10468610 TI - Bacterial inactivation by using near- and supercritical carbon dioxide. AB - The three most common methods of sterilization in use today are ethylene oxide exposure, gamma-irradiation, and steam sterilization. Each of these methods has serious limitations for the sterilization of some materials used in medicine, especially thermally and hydrolytically sensitive polymers by themselves and in combination with proteins. In this work, we demonstrate a potential new method of sterilization by using supercritical fluid carbon dioxide. Using this method we achieve complete inactivation of a wide variety of bacterial organisms at moderate temperatures and in the absence of organic solvents or irradiation. Sterilization is a function of both the proximity to the fluid's critical point and the chemical nature of the fluid itself. When biodegradable polymers poly(lactic-co-glycolic) acid and polylactic acid were included in the sterilization process, there was no effect on the inactivation efficiency, yet no physical or chemical damage to these thermally and hydrolytically labile materials was observed. PMID- 10468611 TI - Correction of the UDP-glucuronosyltransferase gene defect in the gunn rat model of crigler-najjar syndrome type I with a chimeric oligonucleotide. AB - Crigler-Najjar syndrome type I is characterized by unconjugated hyperbilirubinemia resulting from an autosomal recessive inherited deficiency of hepatic UDP-glucuronosyltransferase (UGT) 1A1 activity. The enzyme is essential for glucuronidation and biliary excretion of bilirubin, and its absence can be fatal. The Gunn rat is an excellent animal model of this disease, exhibiting a single guanosine (G) base deletion within the UGT1A1 gene. The defect results in a frameshift and a premature stop codon, absence of enzyme activity, and hyperbilirubinemia. Here, we show permanent correction of the UGT1A1 genetic defect in Gunn rat liver with site-specific replacement of the absent G residue at nucleotide 1206 by using an RNA/DNA oligonucleotide designed to promote endogenous repair of genomic DNA. The chimeric oligonucleotide was either complexed with polyethylenimine or encapsulated in anionic liposomes, administered i.v., and targeted to the hepatocyte via the asialoglycoprotein receptor. G insertion was determined by PCR amplification, colony lift hybridizations, restriction endonuclease digestion, and DNA sequencing, and confirmed by genomic Southern blot analysis. DNA repair was specific, efficient, stable throughout the 6-month observation period, and associated with reduction of serum bilirubin levels. Our results indicate that correction of the UGT1A1 genetic lesion in the Gunn rat restores enzyme expression and bilirubin conjugating activity, with consequent improvement in the metabolic abnormality. PMID- 10468612 TI - The interaction between p53 and DNA topoisomerase I is regulated differently in cells with wild-type and mutant p53. AB - DNA topoisomerase I is a nuclear enzyme involved in transcription, recombination, and DNA damage recognition. Previous studies have shown that topoisomerase I interacts directly with the tumor-suppressor protein p53. p53 is a transcription factor that activates certain genes through binding to specific DNA sequences. We now report that topoisomerase I can be stimulated by both latent and activated wild-type p53 as well as by several mutant and truncated p53 proteins in vitro, indicating that sequence-specific DNA-binding and stimulation of topoisomerase I are distinct properties of p53. These assays also suggest that the binding site for topoisomerase I on p53 is between amino acids 302 and 321. In living cells, the interaction between p53 and topoisomerase I is strongly dependent on p53 status. In MCF-7 cells, which have wild-type p53, the association between the two proteins is tightly regulated in a spatial and temporal manner and takes place only during brief periods of genotoxic stress. In marked contrast, the two proteins are constitutively associated in HT-29 cells, which have mutant p53. These findings have important implications for both cellular stress response and genomic stability, given the ability of topoisomerase I to recognize DNA lesions as well as to cause illegitimate recombination. PMID- 10468613 TI - Expression of oligodendrocyte progenitor cell antigens by gliomas: implications for the histogenesis of brain tumors. AB - The early events in neoplastic transformation can be understood only by comparison of the neoplastic cell with its nontransformed counterpart. The most common central nervous system gliomas traditionally are thought to arise from mature astrocytes and oligodendrocytes. We examined the possibility that gliomas arise from a population of glia that has properties of oligodendrocyte progenitors. These glial cells express the NG2 chondroitin sulfate proteoglycan and the alpha receptor of platelet-derived growth factor in vivo. We identified NG2 and the alpha receptor of platelet-derived growth factor expression in tissue from seven of seven oligodendrogliomas, three of three pilocytic astrocytomas, and one of five glioblastoma multiforme. These data provide evidence that glial tumors arise from glial progenitor cells. Molecules expressed by these progenitor cells should be considered as targets for novel therapeutics. PMID- 10468614 TI - Postexposure immunoprophylaxis of primary isolates by an antibody to HIV receptor complex. AB - mAb B4 is a monoclonal antibody directed against HIV receptor complex. The antibody had broad neutralizing activity against HIV and provided postexposure prophylaxis to hu-peripheral blood leukocyte (PBL)-severe combined immunodeficient mice and chimpanzees. B4 recognized a complex receptor site for HIV on the T cell surface that includes CD4 and also may be influenced by interaction with HIV coreceptors. mAb B4 preferentially neutralized primary HIV-1 isolates compared with T cell line-adapted strains, including syncytium-inducing and non-syncytium-inducing phenotypes, representatives from HIV-1 subtypes A-G, as well as HIV-2, simian immunodeficiency virus, and chimeric simian/human immunodeficiency virus (SHIV). Neutralization was demonstrated in both pre- and postinfection models. The administration of mAb B4 after infectious challenge totally interrupted the infection of hu-PBL-severe combined immunodeficient mice by PBL-grown HIV-1 and the infection of chimpanzees by chimp-adapted HIV-1. This mode of protection suggested that the anti-HIV receptor antibody is efficacious for prophylaxis after exposure to HIV and for prevention of maternal transmission and may be an effective antiretroviral agent for treatment. PMID- 10468615 TI - Comparing the hypothalamic and extrahypothalamic actions of endogenous hyperleptinemia. AB - To determine whether the depletion of body fat caused by adenovirus-induced hyperleptinemia is mediated via the hypothalamus, we used as a "bioassay" for hypothalamic leptin activity the hypothalamic expression of a leptin-regulated peptide, cocaine- and amphetamine-regulated transcript (CART). The validation of this strategy was supported by the demonstration that CART mRNA was profoundly reduced in obese rats with impaired leptin action, whether because of ablation of the ventromedial hypothalamus (VMH) or a loss-of-function mutation in the leptin receptor, as in Zucker diabetic fatty rats. We compared leptin activity in normal rats made hyperleptinemic by adenovirus-leptin treatment (43 +/- 9 ng/ml, cerebrospinal fluid leptin 100 pg/ml) with normal rats made hyperleptinemic by a 60% fat intake (19 +/- 4 ng/ml, cerebrospinal fluid leptin 69 +/- 22 pg/ml). CART was increased 5-fold in the former and 2-fold in the latter, yet in adenovirus induced hyperleptinemia, body fat had disappeared, whereas in high-fat-fed rats, body fat was abundant. Treatment of the high-fat-fed rats with adenovirus-leptin further increased their hyperleptinemia to 56 +/- 6 ng/ml without changing CART mRNA or food intake, indicating that leptin action on hypothalamus had not been increased. Nevertheless, their body fat declined 36%, suggesting that an extrahypothalamic mechanism was responsible. We conclude that in diet-induced obesity body-fat depletion by leptin requires supraphysiologic plasma concentrations that exceed the leptin-transport capacity across the blood-brain barrier. PMID- 10468616 TI - Long-term expression of human coagulation factor VIII and correction of hemophilia A after in vivo retroviral gene transfer in factor VIII-deficient mice. AB - Hemophilia A is caused by a deficiency in coagulation factor VIII (FVIII) and predisposes to spontaneous bleeding that can be life-threatening or lead to chronic disabilities. It is well suited for gene therapy because a moderate increase in plasma FVIII concentration has therapeutic effects. Improved retroviral vectors expressing high levels of human FVIII were pseudotyped with the vesicular stomatitis virus G glycoprotein, were concentrated to high-titers (10(9)-10(10) colony-forming units/ml), and were injected intravenously into newborn, FVIII-deficient mice. High-levels (>/=200 milliunits/ml) of functional human FVIII production could be detected in 6 of the 13 animals, 4 of which expressed physiologic or higher levels (500-12,500 milliunits/ml). Five of the six expressers produced FVIII and survived an otherwise lethal tail-clipping, demonstrating phenotypic correction of the bleeding disorder. FVIII expression was sustained for >14 months. Gene transfer occurred into liver, spleen, and lungs with predominant FVIII mRNA expression in the liver. Six of the seven animals with transient or no detectable human FVIII developed FVIII inhibitors (7 350 Bethesda units/ml). These findings indicate that a genetic disease can be corrected by in vivo gene therapy using retroviral vectors. PMID- 10468618 TI - Activation and adoptive transfer of Epstein-Barr virus-specific cytotoxic T cells in solid organ transplant patients with posttransplant lymphoproliferative disease. AB - The treatment of Epstein-Barr virus (EBV)-associated lymphoproliferative disease (PTLD) in EBV seronegative solid organ transplant recipients who acquire their EBV infection after engraftment poses a considerable challenge because of underlying immunosuppression that inhibits the virus-specific cytotoxic T cell (CTL) response in vivo. We have developed a protocol for activating autologous EBV-specific CTL lines from these patients and show their potential use for immunotherapy against PTLD in solid organ transplant patients. Peripheral blood mononuclear cells from a panel of solid organ transplant recipients with and without active PTLD were used to assess EBV-specific memory CTL responses. The activation protocol involved cocultivation of peripheral blood mononuclear cells with an autologous lymphoblastoid cell line under conditions that favored expansion of virus-specific CTL and hindered the proliferation of allospecific T cells. These CTL consistently showed (i) strong EBV-specificity, including reactivity through defined epitopes in spite of concurrent immunosuppressive therapy, and (ii) no alloreactivity toward donor alloantigens. More importantly, adoptive transfer of these autologous CTLs into a single patient with active PTLD was coincident with a very significant regression of the PTLD. These results demonstrate that a potent EBV-specific memory response can be expanded from solid organ recipients who have acquired their primary EBV infection under high levels of immunosuppressive therapy and that these T cells may have therapeutic potential against PTLD. PMID- 10468617 TI - Genetic alteration of phospholipase C beta3 expression modulates behavioral and cellular responses to mu opioids. AB - Morphine and other micro opioids regulate a number of intracellular signaling pathways, including the one mediated by phospholipase C (PLC). By studying PLC beta3-deficient mice, we have established a strong link between PLC and mu opioid mediated responses at both the behavioral and cellular levels. Mice lacking PLC beta3, when compared with the wild type, exhibited up to a 10-fold decrease in the ED(50) value for morphine in producing antinociception. The reduced ED(50) value was unlikely a result of changes in opioid receptor number or affinity because no differences were found in whole-brain B(max) and K(d) values for mu, kappa, and delta opioid receptors between wild-type and PLC beta3-null mice. We also found that opioid regulation of voltage-sensitive Ca(2+) channels in primary sensory neurons (dorsal root ganglion) was different between the two genotypes. Consistent with the behavioral findings, the specific mu agonist [D Ala(2),(Me)Phe(4),Gly(ol)(5)]enkephalin (DAMGO) induced a greater whole-cell current reduction in a greater proportion of neurons isolated from the PLC beta3 null mice than from the wild type. In addition, reconstitution of recombinant PLC protein back into PLC beta3-deficient dorsal root ganglion neurons reduced DAMGO responses to those of wild-type neurons. In neurons of both genotypes, activation of protein kinase C with phorbol esters markedly reduced DAMGO-mediated Ca(2+) current reduction. These data demonstrate that PLC beta3 constitutes a significant pathway involved in negative modulation of mu opioid responses, perhaps via protein kinase C, and suggests the possibility that differences in opioid sensitivity among individuals could be, in part, because of genetic factors. PMID- 10468619 TI - Vasopressin contributes to hyperfiltration, albuminuria, and renal hypertrophy in diabetes mellitus: study in vasopressin-deficient Brattleboro rats. AB - Diabetic nephropathy represents a major complication of diabetes mellitus (DM), and the origin of this complication is poorly understood. Vasopressin (VP), which is elevated in type I and type II DM, has been shown to increase glomerular filtration rate in normal rats and to contribute to progression of chronic renal failure in 5/6 nephrectomized rats. The present study was thus designed to evaluate whether VP contributes to the renal disorders of DM. Renal function was compared in Brattleboro rats with diabetes insipidus (DI) lacking VP and in normal Long-Evans (LE) rats, with or without streptozotocin-induced DM. Blood and urine were collected after 2 and 4 weeks of DM, and creatinine clearance, urinary glucose and albumin excretion, and kidney weight were measured. Plasma glucose increased 3-fold in DM rats of both strains, but glucose excretion was approximately 40% lower in DI-DM than in LE-DM, suggesting less intense metabolic disorders. Creatinine clearance increased significantly in LE-DM (P < 0.01) but failed to increase in DI-DM. Urinary albumin excretion more than doubled in LE-DM but rose by only 34% in DI-DM rats (P < 0.05). Kidney hypertrophy was also less intense in DI-DM than in LE-DM (P < 0.001). These results suggest that VP plays a critical role in diabetic hyperfiltration and albuminuria induced by DM. This hormone thus seems to be an additional risk factor for diabetic nephropathy and, thus, a potential target for prevention and/or therapeutic intervention. PMID- 10468620 TI - Epoxomicin, a potent and selective proteasome inhibitor, exhibits in vivo antiinflammatory activity. AB - The proteasome regulates cellular processes as diverse as cell cycle progression and NF-kappaB activation. In this study, we show that the potent antitumor natural product epoxomicin specifically targets the proteasome. Utilizing biotinylated-epoxomicin as a molecular probe, we demonstrate that epoxomicin covalently binds to the LMP7, X, MECL1, and Z catalytic subunits of the proteasome. Enzymatic analyses with purified bovine erythrocyte proteasome reveal that epoxomicin potently inhibits primarily the chymotrypsin-like activity. The trypsin-like and peptidyl-glutamyl peptide hydrolyzing catalytic activities also are inhibited at 100- and 1,000-fold slower rates, respectively. In contrast to peptide aldehyde proteasome inhibitors, epoxomicin does not inhibit nonproteasomal proteases such trypsin, chymotrypsin, papain, calpain, and cathepsin B at concentrations of up to 50 microM. In addition, epoxomicin is a more potent inhibitor of the chymotrypsin-like activity than lactacystin and the peptide vinyl sulfone NLVS. Epoxomicin also effectively inhibits NF-kappaB activation in vitro and potently blocks in vivo inflammation in the murine ear edema assay. These results thus define epoxomicin as a novel proteasome inhibitor that likely will prove useful in exploring the role of the proteasome in various in vivo and in vitro systems. PMID- 10468621 TI - Role of the type 5 adenovirus gene encoding the early region 1B 55-kDa protein in pulmonary pathogenesis. AB - Comparison of the inflammatory response of Sigmodon hispidus cotton rats to pulmonary infection with wild-type 5 adenovirus (Ad5) or with a viral mutant, in which the early region 1B gene encoding a 55-kDa protein, Ad5dl110 (dl110), was deleted, indicated that the inflammation in animals infected with dl110 was markedly reduced compared with the inflammation in animals infected with wild type Ad5, although both viruses replicated to the same extent. Comparable experiments done with C57BL/6 mice yielded identical results, even though only the early phase of gene expression essential for viral replication occurs in mice. Cytokine analysis of infected mouse lungs indicated that tumor necrosis factor-alpha and IL-6 were produced in relatively large quantities in wild-type Ad5-infected mice and at significantly lower levels in dl110-infected mice during the early stages of infection. PMID- 10468622 TI - Bacillus subtilis aconitase is an RNA-binding protein. AB - The aconitase protein of Bacillus subtilis was able to bind specifically to sequences resembling the iron response elements (IREs) found in eukaryotic mRNAs. The sequences bound include the rabbit ferritin IRE and IRE-like sequences in the B. subtilis operons that encode the major cytochrome oxidase and an iron uptake system. IRE binding activity was affected by the availability of iron both in vivo and in vitro. In eukaryotic cells, aconitase-like proteins regulate translation and stability of iron metabolism mRNAs in response to iron availability. A mutant strain of B. subtilis that produces an enzymatically inactive aconitase that was still able to bind RNA sporulated 40x more efficiently than did an aconitase null mutant, suggesting that a nonenzymatic activity of aconitase is important for sporulation. The results support the idea that bacterial aconitases, like their eukaryotic homologs, are bifunctional proteins, showing aconitase activity in the presence of iron and RNA binding activity when cells are iron-deprived. PMID- 10468623 TI - Where is my arm? The relative role of vision and proprioception in the neuronal representation of limb position. AB - A central problem in motor control, in the representation of space, and in the perception of body schema is how the brain encodes the relative positions of body parts. According to psychophysical studies, this sense of limb position depends heavily on vision. However, almost nothing is currently known about how the brain uses vision to determine or represent the location of the arm or any other body part. The present experiment shows that the position of the arm is represented in the premotor cortex of the monkey (Macaca fascicularis) brain by means of a convergence of visual cues and proprioceptive cues onto the same neurons. These neurons respond to the felt position of the arm when the arm is covered from view. They also respond in a similar fashion to the seen position of a false arm. PMID- 10468624 TI - Tracking neuronal fiber pathways in the living human brain. AB - Functional imaging with positron emission tomography and functional MRI has revolutionized studies of the human brain. Understanding the organization of brain systems, especially those used for cognition, remains limited, however, because no methods currently exist for noninvasive tracking of neuronal connections between functional regions [Crick, F. & Jones, E. (1993) Nature (London) 361, 109-110]. Detailed connectivities have been studied in animals through invasive tracer techniques, but these invasive studies cannot be done in humans, and animal results cannot always be extrapolated to human systems. We have developed noninvasive neuronal fiber tracking for use in living humans, utilizing the unique ability of MRI to characterize water diffusion. We reconstructed fiber trajectories throughout the brain by tracking the direction of fastest diffusion (the fiber direction) from a grid of seed points, and then selected tracks that join anatomically or functionally (functional MRI) defined regions. We demonstrate diffusion tracking of fiber bundles in a variety of white matter classes with examples in the corpus callosum, geniculo-calcarine, and subcortical association pathways. Tracks covered long distances, navigated through divergences and tight curves, and manifested topological separations in the geniculo-calcarine tract consistent with tracer studies in animals and retinotopy studies in humans. Additionally, previously undescribed topologies were revealed in the other pathways. This approach enhances the power of modern imaging by enabling study of fiber connections among anatomically and functionally defined brain regions in individual human subjects. PMID- 10468625 TI - Widespread accumulation of [(3)H]testosterone in the spinal cord of a wild bird with an elaborate courtship display. AB - Elaborate courtship displays are relatively common features of the masculine reproductive behavior in birds. However, little is known about their neural and hormonal control. One bird that performs such a display is the golden-collared manakin (Manacus vitellinus) of Panamanian forests. Adult males, but not females, perform a physically intense display requiring substantial neuromuscular control of the wings and legs. We tested the hypothesis that steroid sensitivity is a property of neurons in the manakin spinal cord. Males and females were captured from active courtship leks, treated with drugs to block steroidogenesis, injected with (3)H-labeled testosterone, and the spinal cords were removed and processed for autoradiography. Sex steroid-accumulating cells were widely distributed in the spinal cords in each of six males and in one of five females. Cells, including presumptive motoneurons, reached their highest density in the ventral horns of the cervical and lumbosacral enlargements, regions associated with motor control of the wings and legs. These results suggest that neurons in the adult manakin spinal cord can express sex-steroid receptors, but do so less in females than in males. This evidence for androgen sensitivity and sexual dimorphism in the adult avian spinal cord suggests that sex steroids may control diverse behaviors in male birds in part by acting directly on the spinal neural circuits. PMID- 10468626 TI - Volatile anesthetics block actin-based motility in dendritic spines. AB - Dendritic spines form the postsynaptic contact sites for most excitatory synapses in the brain. Spines occur in a wide range of different shapes that can vary depending on an animal's experience or behavioral status. Recently we showed that spines on living neurons can change shape within seconds in a process that depends on actin polymerization. We have now found that this morphological plasticity is blocked by inhalational anesthetics at concentrations at which they are clinically effective. These volatile compounds also block actin-based motility in fibroblasts, indicating that their action is independent of neuron specific components and thus identifying the actin cytoskeleton as a general cellular target of anesthetic action. These observations imply that inhibition of actin dynamics at brain synapses occurs during general anesthesia and that inhalational anesthetics are capable of influencing the morphological plasticity of excitatory synapses in the brain. PMID- 10468627 TI - Dynamic visualization of nervous system in live Drosophila. AB - We have constructed transgenic Drosophila melanogaster lines that express green fluorescent protein (GFP) exclusively in the nervous system. Expression is controlled with transcriptional regulatory elements present in the 5' flanking DNA of the Drosophila Na(+), K(+)-ATPase beta-subunit gene Nervana2 (Nrv2). This regulatory DNA is fused to the yeast transcriptional activator GAL4, which binds specifically to a sequence motif termed the UAS (upstream activating sequence). Drosophila lines carrying Nrv2-GAL4 transgenes have been genetically recombined with UAS-GFP (S65T) transgenes (Nrv2-GAL4+UAS-GFP) inserted on the same chromosomes. We observe strong nervous system-specific fluorescence in embryos, larvae, pupae, and adults. The GFP fluorescence is sufficiently bright to allow dynamic imaging of the nervous system at all of these developmental stages directly through the cuticle of live Drosophila. These lines provide an unprecedented view of the nervous system in living animals and will be valuable tools for investigating a number of developmental, physiological, and genetic neurobiological problems. PMID- 10468628 TI - Targeted disruption of the orphanin FQ/nociceptin gene increases stress susceptibility and impairs stress adaptation in mice. AB - The neuropeptide orphanin FQ (also known as nociceptin; OFQ/N) has been implicated in modulating stress-related behavior. OFQ/N was demonstrated to reverse stress-induced analgesia and possess anxiolytic-like activity after central administration. To further study physiological functions of OFQ/N, we have generated OFQ/N-deficient mice by targeted disruption of the OFQ/N gene. Homozygous mice display increased anxiety-like behavior when exposed to a novel and threatening environment. OFQ/N-null mice show elevated basal pain threshold but develop normal stress-induced analgesia. Interestingly, these mice show impaired adaptation to repeated stress when compared with wild-type mice, whereas their performance in spatial learning remained unaffected. Basal and poststress plasma corticosterone levels were found to be elevated in OFQ/N-deficient animals. Thus, OFQ/N appears to be crucially involved in the neurobiological regulation of stress-coping behavior and fear. PMID- 10468629 TI - Noise shaping in populations of coupled model neurons. AB - Biological information-processing systems, such as populations of sensory and motor neurons, may use correlations between the firings of individual elements to obtain lower noise levels and a systemwide performance improvement in the dynamic range or the signal-to-noise ratio. Here, we implement such correlations in networks of coupled integrate-and-fire neurons using inhibitory coupling and demonstrate that this can improve the system dynamic range and the signal-to noise ratio in a population rate code. The improvement can surpass that expected for simple averaging of uncorrelated elements. A theory that predicts the resulting power spectrum is developed in terms of a stochastic point-process model in which the instantaneous population firing rate is modulated by the coupling between elements. PMID- 10468630 TI - Linguistic threat activates the human amygdala. AB - Studies in animals demonstrate a crucial role for the amygdala in emotional and social behavior, especially as related to fear and aggression. Whereas lesion and functional-imaging studies in humans indicate the amygdala's participation in assessing the significance of nonverbal as well as paralinguistic cues, direct evidence for its role in the emotional processing of linguistic cues is lacking. In this study, we use a modified Stroop task along with a high-sensitivity neuroimaging technique to target the neural substrate engaged specifically when processing linguistic threat. Healthy volunteer subjects were instructed to name the color of words of either threat or neutral valence, presented in different color fonts, while neural activity was measured by using H(2)(15)O positron emission tomography. Bilateral amygdalar activation was significantly greater during color naming of threat words than during color naming of neutral words. Associated activations were also noted in sensory-evaluative and motor-planning areas of the brain. Thus, our results demonstrate the amygdala's role in the processing of danger elicited by language. In addition, the results reinforce the amygdala's role in the modulation of the perception of, and response to, emotionally salient stimuli. The current study further suggests conservation of phylogenetically older mechanisms of emotional evaluation in the context of more recently evolved linguistic function. PMID- 10468631 TI - Native language, gender, and functional organization of the auditory cortex. AB - Whole-head magnetoencephalography was employed in 40 normal subjects to investigate whether the basic functional organization of the auditory cortex varies with linguistic environment. Robust activations of the bilateral supratemporal auditory cortices to 1-kHz pure tones, maximum at about 100 ms after stimulus onset, were studied in Finnish and German female and male subject groups with monolingual background. Activations elicited by the tones were mutually indistinguishable in German and Finnish women. In contrast, German men showed significantly stronger auditory responses to pure tones in the left, language-dominant hemisphere than Finnish men. We discuss the possibility that the prominent left-hemisphere activation in German males reflects higher frequency resolution required for distinguishing between German than Finnish vowels and that the clear effect of native language in male but not in female auditory cortex derives from more pronounced functional lateralization in men. The present data suggest that the influence of native language can extend to auditory cortical processing of pure-tone stimuli with no linguistic content and that this effect is conspicuous in the male brain. PMID- 10468632 TI - Serum choline activates mutant acetylcholine receptors that cause slow channel congenital myasthenic syndromes. AB - We have found that mutant acetylcholine receptor channels (AChRs) that cause slow channel congenital myasthenic syndromes are activated by serum and that the high frequency of openings in serum is reduced by treatment with choline oxidase. Thus, slow-channel congenital myasthenic syndrome AChRs at the neuromuscular junction are likely to be activated both by steady exposure to serum choline and by transient exposure to synaptically released transmitter. Single-channel kinetic analyses indicate that the increased response to choline is caused by a reduced intrinsic stability of the closed channel. The results suggest that a mutation that destabilizes the inactive conformation of the AChR, together with the sustained exposure of endplates to serum choline, results in continuous channel activity that contributes to the pathophysiology of the disease. PMID- 10468633 TI - Distinct human NUMB isoforms regulate differentiation vs. proliferation in the neuronal lineage. AB - Neuronal cell fate decisions are directed in Drosophila by NUMB, a signaling adapter protein with two protein-protein interaction domains: a phosphotyrosine binding domain and a proline-rich region (PRR) that functions as an SH3-binding domain. Here we show that there are at least four human NUMB isoforms and that these serve two distinct developmental functions in the neuronal lineage: differentiation (but not proliferation) is promoted by human NUMB protein isoforms with a type I (short) PRR. In contrast, proliferation (but not differentiation) is directed by isoforms that have a type II (long) PRR. The two types of PRR may promote distinct intracellular signaling pathways downstream of the NOTCH receptor during mammalian neurogenesis. PMID- 10468634 TI - Thermal avoidance in Caenorhabditis elegans: an approach to the study of nociception. AB - Upon perception of a noxious stimulus, an organism executes defense mechanisms, such as escape responses. The molecular basis of these mechanisms is poorly understood. In this paper we show that upon exposure to noxious temperature, Caenorhabditis elegans reacts by a withdrawal reflex. To analyze this thermal avoidance behavior, we developed a laser-based assay to quantify the response. The escape reflex can be observed in 98% of the adult animals, but is not executed in animals in diapause. The thermal avoidance response differs significantly from the thermotaxis behavior that is based on the perception of physiological temperature. It involves different neurons and is influenced by mutations in distinct genes. As in mammals, the strength of the thermal avoidance response is increased by application of capsaicin, the pungent ingredient in chili peppers. We find that thermal avoidance is strongly reduced in mutants affecting the neural transmission modulated by glutamate and neuropeptides as well as in mutants affecting the structure and function of sensory neurons. We suggest that the study of this nociceptive behavior in C. elegans can be used to understand the genetic and molecular basis of thermal nociception. PMID- 10468635 TI - Enhancement of D1 dopamine receptor-mediated locomotor stimulation in M(4) muscarinic acetylcholine receptor knockout mice. AB - Muscarinic acetylcholine receptors (M(1)-M(5)) regulate many key functions of the central and peripheral nervous system. Primarily because of the lack of receptor subtype-selective ligands, the precise physiological roles of the individual muscarinic receptor subtypes remain to be elucidated. Interestingly, the M(4) receptor subtype is expressed abundantly in the striatum and various other forebrain regions. To study its potential role in the regulation of locomotor activity and other central functions, we used gene-targeting technology to create mice that lack functional M(4) receptors. Pharmacologic analysis of M(4) receptor deficient mice indicated that M(4) receptors are not required for muscarinic receptor-mediated analgesia, tremor, hypothermia, and salivation. Strikingly, M(4) receptor-deficient mice showed an increase in basal locomotor activity and greatly enhanced locomotor responses (as compared with their wild-type littermates) after activation of D1 dopamine receptors. These results indicate that M(4) receptors exert inhibitory control on D1 receptor-mediated locomotor stimulation, probably at the level of striatal projection neurons where the two receptors are coexpressed at high levels. Our findings offer new perspectives for the treatment of Parkinson's disease and other movement disorders that are characterized by an imbalance between muscarinic cholinergic and dopaminergic neurotransmission. PMID- 10468636 TI - Mitomycin resistance in mammalian cells expressing the bacterial mitomycin C resistance protein MCRA. AB - The mitomycin C-resistance gene, mcrA, of Streptomyces lavendulae produces MCRA, a protein that protects this microorganism from its own antibiotic, the antitumor drug mitomycin C. Expression of the bacterial mcrA gene in mammalian Chinese hamster ovary cells causes profound resistance to mitomycin C and to its structurally related analog porfiromycin under aerobic conditions but produces little change in drug sensitivity under hypoxia. The mitomycins are prodrugs that are enzymatically reduced and activated intracellularly, producing cytotoxic semiquinone anion radical and hydroquinone reduction intermediates. In vitro, MCRA protects DNA from cross-linking by the hydroquinone reduction intermediate of these mitomycins by oxidizing the hydroquinone back to the parent molecule; thus, MCRA acts as a hydroquinone oxidase. These findings suggest potential therapeutic applications for MCRA in the treatment of cancer with the mitomycins and imply that intrinsic or selected mitomycin C resistance in mammalian cells may not be due solely to decreased bioactivation, as has been hypothesized previously, but instead could involve an MCRA-like mechanism. PMID- 10468637 TI - Disruption of myoglobin in mice induces multiple compensatory mechanisms. AB - Myoglobin may serve a variety of functions in muscular oxygen supply, such as O(2) storage, facilitated O(2) diffusion, and myoglobin-mediated oxidative phosphorylation. We studied the functional consequences of a myoglobin deficiency on cardiac function by producing myoglobin-knockout (myo(-/-)) mice. To genetically inactivate the myoglobin gene, exon 2 encoding the heme binding site was deleted in embryonic stem cells via homologous recombination. Myo(-/-) mice are viable, fertile, and without any obvious signs of functional limitations. Hemoglobin concentrations were significantly elevated in myo(-/-) mice. Cardiac function and energetics were analyzed in isolated perfused hearts under resting conditions and during beta-adrenergic stimulation with dobutamine. Myo(-/-) hearts showed no alteration in contractile parameters either under basal conditions or after maximal beta-adrenergic stimulation (200 nM dobutamine). Tissue levels of ATP, phosphocreatine ((31)P-NMR), and myocardial O(2) consumption were not altered. However, coronary flow [6.4 +/- 1.3 ml.min(-1).g( 1) [wild-type (WT)] vs. 8.5 +/- 2.4 ml.min(-1).g(-1) [myo(-/-)] [and coronary reserve [17.1 +/- 2.1 (WT) vs. 20.8 +/- 1.1 (myo(-/-) ml. min(-1).g(-1) were significantly elevated in myo(-/-) hearts. Histological examination revealed that capillary density also was increased in myo(-/-) hearts [3,111 +/- 400 mm(-2) (WT) vs. 4,140 +/- 140 mm(-2) (Myo(-/-)]. These data demonstrate that disruption of myoglobin results in the activation of multiple compensatory mechanisms that steepen the pO(2) gradient and reduce the diffusion path length for O(2) between capillary and the mitochondria; this suggests that myoglobin normally is important for the delivery of oxygen. PMID- 10468638 TI - Abortive expansion of the cumulus and impaired fertility in mice lacking the prostaglandin E receptor subtype EP(2). AB - Female mice lacking the gene encoding the prostaglandin (PG) E(2) receptor subtype EP(2) (EP(2)(-/-)) become pregnant and deliver their pups at term, but with a much reduced litter size. A decrease in ovulation number and a much reduced fertilization rate were observed in EP(2)(-/-) females without difference of the uterus to support implantation of wild-type embryos. Treatment with gonadotropins induced EP(2) mRNA expression in the cumulus cells of ovarian follicles of wild-type mice. The immature cumuli oophori from wild-type mice expanded in vitro in response to both follicle-stimulating hormone and PGE(2), but the response to PGE(2) was absent in those from EP(2)(-/-) mice. Cumulus expansion proceeded normally in preovulatory follicles but became abortive in a number of ovulated complexes in EP(2)(-/-) mice, indicating that EP(2) is involved in cumulus expansion in the oviduct in vivo. No difference in the fertilization rate between wild-type and EP(2)(-/-) mice was found in in vitro studies using cumulus-free oocytes. These results indicate that PGE(2) cooperates with gonadotropin to complete cumulus expansion for successful fertilization. PMID- 10468639 TI - The AtCAO gene, encoding chlorophyll a oxygenase, is required for chlorophyll b synthesis in Arabidopsis thaliana. AB - Chlorophyll b is synthesized from chlorophyll a and is found in the light harvesting complexes of prochlorophytes, green algae, and both nonvascular and vascular plants. We have used conserved motifs from the chlorophyll a oxygenase (CAO) gene from Chlamydomonas reinhardtii to isolate a homologue from Arabidopsis thaliana. This gene, AtCAO, is mutated in both leaky and null chlorina1 alleles, and DNA sequence changes cosegregate with the mutant phenotype. AtCAO mRNA levels are higher in three different mutants that have reduced levels of chlorophyll b, suggesting that plants that do not have sufficient chlorophyll b up-regulate AtCAO gene expression. Additionally, AtCAO mRNA levels decrease in plants that are grown under dim-light conditions. We have also found that the six major Lhcb proteins do not accumulate in the null ch1-3 allele. PMID- 10468640 TI - Sugars modulate an unusual mode of control of the cell-wall invertase gene (Incw1) through its 3' untranslated region in a cell suspension culture of maize. AB - We show here that a cell-wall invertase encoded by the Incw1 gene is regulated at both the transcriptional and posttranscriptional levels by sugars in a heterotrophic cell suspension culture of maize. The Incw1 gene encoded two transcripts: Incw1-S (small) and Incw1-L (large); the size variation was attributable to different lengths in the 3' untranslated region. Both metabolizable and nonmetabolizable sugars induced Incw1-L RNA apparently by default. However, only the metabolizable sugars, sucrose and D-glucose, were associated with the increased steady-state abundance of Incw1-S RNA, the concomitant increased levels of INCW1 protein and enzyme activity, and the downstream metabolic repression of the sucrose synthase gene, Sh1. Conversely, nonmetabolizable sugars, including the two glucose analogs 3-O-methylglucose and 2-deoxyglucose, induced greater steady-state levels of the Incw1-L RNA, but this increase did not lead to either an increase in the levels of the INCW1 protein/enzyme activity or the repression of the Sh1 gene. We conclude that sugar sensing and the induction of the Incw1 gene is independent of the hexokinase pathway. More importantly, our results also suggest that the 3' untranslated region of the Incw1 gene acts as a regulatory sensor of carbon starvation and may constitute a link between sink metabolism and cellular translation in plants. PMID- 10468641 TI - Analysis of population structure of the chestnut blight fungus based on vegetative incompatibility genotypes. AB - Vegetative incompatibility is a self/nonself-recognition system in fungi that has often been used for describing phenotypic diversity in fungal populations. A common hypothesis is that vegetative incompatibility polymorphisms are maintained by balancing selection. However, understanding the evolutionary significance of vegetative incompatibility and the factors that maintain these polymorphisms has been limited by a lack of knowledge of the underlying genetics of vegetative compatibility (vc) types. Genotypes of 64 vc types, controlled by six unlinked vegetative incompatibility (vic) loci, have been identified in the chestnut blight fungus, Cryphonectria parasitica. By interpreting vc type survey data in terms of vic genotypes, we estimated vic-allele frequencies and analyzed the multilocus genetic structure of 13 populations in Europe and 3 populations in the U.S. European populations have less vc type diversity than the US populations because of a combination of lower vic-allele diversity and limited recombination. Genotypic diversity of 10 populations in Italy correlated to the abundance of sexual structures; however, significant deviations from random mating suggest that either sexual reproduction may not contribute many offspring in these populations or that vic genes (or vic genotypes) are under selection. Most vic allele frequencies deviated from 0.5, the equilibrium frequency predicted under frequency-dependent selection, providing no evidence for selection acting on these loci. PMID- 10468642 TI - Associative plasticity in striatal transplants. AB - Striatal lesions disrupt both motor and cognitive performance in rats, many aspects of which can be restored by striatal transplants. Because the normal striatum is involved in the formation and maintenance of motor habits, it has been hypothesized that grafted animals may require explicit retraining to relearn previously established habits that have been disrupted by the lesions. We have used a lateralized-discrimination task to reproduce this "learning to use the transplant" effect, combined with a transfer-of-training paradigm to demonstrate that recovery requires relearning specific lateralized stimulus-response associations and cannot be explained simply by a generalized training-dependent improvement in motor skill. These results have clear implications for developing appropriate strategies for the rehabilitation of Huntington's disease patients participating in clinical transplantation programs. PMID- 10468643 TI - Contextual influences in V1 as a basis for pop out and asymmetry in visual search. AB - I use a model to show how simple, bottom-up, neural mechanisms in primary visual cortex can qualitatively explain the preattentive component of complex psychophysical phenomena of visual search for a target among distracters. Depending on the image features, the speed of search ranges from fast, when a target pops-out or is instantaneously detectable, to very slow, and it can be asymmetric with respect to switches between the target and distracter objects. It has been unclear which neural mechanisms or even cortical areas control the ease of search, and no physiological correlate has been found for search asymmetry. My model suggests that contextual influences in V1 play a significant role. PMID- 10468644 TI - Natural experimental models: the global search for biomedical paradigms among traditional, modernizing, and modern populations. AB - During the past four decades, biomedical scientists have slowly begun to recognize the unique opportunities for studying biomedical processes, disease etiology, and mechanisms of pathogenesis in populations with unusual genetic structures, physiological characteristics, focal endemic disease, or special circumstances. Such populations greatly extend our research capabilities and provide a natural laboratory for studying relationships among biobehavioral, genetic, and ecological processes that are involved in the development of disease. The models presented illustrate three different types of natural experiments: those occurring in traditionally living, modernizing, and modern populations. The examples are drawn from current research that involves population mechanisms of adaptation among East African Turkana pastoralists; a search for etiology and mechanisms of pathogenesis of an emerging disease among the Yakut people of Siberia; and psychosocial stress, hypertension, and cardiovascular disease in women working outside the home in New York City and among subpopulations in Hawaii. The models in general, and the examples in specific, represent natural laboratories in which relatively small intrapopulation differences and large interpopulation differences can be used to evaluate health and disease outcomes. PMID- 10468645 TI - Evolutionary demographic models for mortality plateaus. AB - Plateaus in the age pattern of hazard functions at extreme ages have been discovered in large populations of medflies, Drosophila, nematodes, and people. Mueller and Rose [(1996) Proc. Natl. Acad. Sci. USA 93, 15249-15253] have proposed several age-structured demographic models to represent effects of mutation accumulation and antagonistic pleiotropy on randomly evolving schedules of demographic rates. They assert that "evolutionary theory [as embodied in their models] predicts late-life mortality plateaus." This paper defines a class of Markovian models that includes those of Mueller and Rose and obtains a characterization of the possible limiting states. For the basic model, the result implies that schedules with late-life mortality plateaus above a minimal threshold are not limiting states. The models fail, but not for reasons previously conjectured. Transient states, visited early by the process, do display mortality plateaus. Other models from this class may have a role to play in reconciling observed plateaus with evolutionary theory. PMID- 10468646 TI - The treatment of hepatitis C: emerging evidence of the need for change. PMID- 10468647 TI - Hepatitis C: epidemiology and review of complementary/alternative medicine treatments. AB - Hepatitis C is emerging as a serious worldwide problem. In the United States the current mortality figures may triple in the next ten years, rivaling HIV. The disease has a latency of 10-30 years and symptoms or signs may not appear until cirrhosis is evident. Adequate diagnosis, including liver biopsy, is essential in assessing the current stage of the viral infection and the need for treatment. Hepatitis C may manifest as hepatic fibrosis, cirrhosis, hepatocellular carcinoma, lichen planus, glomerulonephritis, mixed cryoglobulinemia, or porphyria. The hepatic damage is due both to the cytopathic effect of the virus and the inflammatory changes secondary to immune activation. The use of the botanical components glycyrrhizin, catechin, silymarin and phytosterols, and the antioxidants N-acetylcysteine and vitamin E are reviewed for their efficacy in treating chronic hepatitis and affecting liver damage. PMID- 10468648 TI - Therapeutic considerations of L-glutamine: a review of the literature. AB - The most abundant amino acid in the bloodstream, L-glutamine fulfills a number of biochemical needs. It operates as a nitrogen shuttle, taking up excess ammonia and forming urea. It can contribute to the production of other amino acids, glucose, nucleotides, protein, and glutathione. Glutamine is primarily formed and stored in skeletal muscle and lungs, and is the principal metabolic fuel for small intestine enterocytes, lymphocytes, macrophages, and fibroblasts. Supplemental use of glutamine, either in oral, enteral, or parenteral form, increases intestinal villous height, stimulates gut mucosal cellular proliferation, and maintains mucosal integrity. It also prevents intestinal hyperpermeability and bacterial translocation, which may be involved in sepsis and the development of multiple organ failure. L-glutamine use has been found to be of great importance in the treatment of trauma and surgery patients, and has been shown to decrease the incidence of infection in these patients. Cancer patients often develop muscle glutamine depletion, due to uptake by tumors and chronic protein catabolism. Glutamine may be helpful in offsetting this depletion; however, it may also stimulate the growth of some tumors. The use of glutamine with cancer chemotherapy and radiotherapy seems to prevent gut and oral toxic side-effects, and may even increase the effectiveness of some chemotherapy drugs. PMID- 10468649 TI - Nutritional and botanical interventions to assist with the adaptation to stress. AB - Prolonged stress, whether a result of mental/emotional upset or due to physical factors such as malnutrition, surgery, chemical exposure, excessive exercise, sleep deprivation, or a host of other environmental causes, results in predictable systemic effects. The systemic effects of stress include increased levels of stress hormones such as cortisol, a decline in certain aspects of immune system function such as natural killer cell cytotoxicity or secretory-IgA levels, and a disruption of gastrointestinal microflora balance. These systemic changes might be a substantial contributor to many of the stress-associated declines in health. Based on human and animal research, it appears a variety of nutritional and botanical substances - such as adaptogenic herbs, specific vitamins including ascorbic acid, vitamins B1 and B6, the coenzyme forms of vitamin B5 (pantethine) and B12 (methylcobalamin), the amino acid tyrosine, and other nutrients such as lipoic acid, phosphatidylserine, and plant sterol/sterolin combinations - may allow individuals to sustain an adaptive response and minimize some of the systemic effects of stress. PMID- 10468650 TI - Comparative measurements of serum estriol, estradiol, and estrone in non pregnant, premenopausal women; a preliminary investigation. AB - Little to no data exists in the literature for serum estriol values in non pregnant, premenopausal women. The current medical community opinion holds that estriol has no significant role in non-pregnant women relative to the other estrogens. It is a possibility that estriol's primary function has yet to be discovered. Accordingly, the first step is to understand cycle-dependent serum estriol concentrations. We have made a preliminary investigation for serum estriol concentration of 26 women during the known cycle peaks of estrone and estradiol. Five of the women were also tested for serum estriol on various days throughout the cycle in order to develop a cycle-dependent concentration profile. The result of these experiments show that serum estriol was always significantly higher than the sum of estrone and estradiol and less fluctuating. We conclude that estriol is probably a significant estrogen component. PMID- 10468651 TI - Silybum marianum (milk thistle). PMID- 10468652 TI - Mechanisms for activation of aortic baroreceptor C-fibres in rabbits and rats. AB - In an earlier study, we examined the pressure-response characteristics of rat aortic baroreceptors with C-fibre (non-medullated) afferents. Compared with aortic baroreceptor fibres with A-fibre (medullated) afferents, the C-fibres were activated at higher pressures and discharged more irregularly when stimulated with a steady level of pressure. Here we examine the relationship between discharge and the aortic diameter in these two types of afferents in rats and rabbits. An in vitro aortic arch/aortic nerve preparation was used to record single-fibre activity simultaneously with aortic arch pressure and diameter. Diameter was measured using a highly sensitive non-contact photoelectric device. Baroreceptor discharge was characterized by stimulating the nerve endings with either slow pressure ramps from subthreshold to 200-250 mmHg, at a rate of rise of 2 mmHg s-1, or pressure steps from subthreshold to suprathreshold levels, at amplitudes of 110-180 mmHg. In response to these inputs, C-fibres in rabbits (conduction velocities= 0.8-2.2 m s-1) behaved much like those in rats. The C fibres had significantly higher pressure thresholds (95 +/- 3 mmHg vs. 53 +/- 2 mmHg; mean +/- SEM), lower threshold frequencies (2.4 +/- 0.5 vs. 27.7 +/- 1.8 spikes s-1), lower maximum discharge frequencies (22.7 +/- 2.3 vs. 65 +/- 5.8 spikes s-1) and more irregular discharge in response to a pressure step when compared with A-fibres (conduction velocities of 8-16 m s-1). When plotted against diameter, C-fibre ramp-evoked discharge increased gradually at first, and then rose steeply at increasingly higher ramp pressures where aortic diameter became relatively constant. In contrast, A-fibre discharge was linearly related to diameter over a wide range of pressure. These results suggest two interpretations: (1) The relation between stretch and C-fibre discharge is highly non-linear, with a marked increase in sensitivity at large diameters. (2) C fibres are stimulated by changes in intramural stress rather than stretch. PMID- 10468653 TI - Calcium response to adenosine and ATP in rabbit afferent arterioles. AB - The effects of purine compounds in the renal vasculature are almost exclusively restricted to pre-glomerular vessels. Although their physiological role as extracellular messengers is not clear, there are extensive data indicating the importance of adenosine and ATP in the regulation of renal haemodynamics. This study was undertaken to characterize the calcium response of rabbit afferent arteriole to adenosine, ATP and other nucleotides. Experiments were performed in isolated afferent arterioles, microdissected from rabbit kidneys and loaded with fura-2. Intracellular calcium concentration ([Ca2+]i) was measured by video in proximal and distal parts of the afferent arteriole. Application of 100 microM adenosine or ATP increased [Ca2+]i in both arteriolar regions. In all cases the response had two well distinguishable phases: a quick peak increase and a plateau phase that equilibrated at a [Ca2+]i significantly higher than the basal level. UTP (100 microM) had no effect on the arteriole. Removal of extracellular calcium (2.5 mM EGTA) abolished only the plateau phase in response to adenosine, without significantly changing the peak increase. In contrast, the response to ATP was completely abolished in both arteriolar regions, where [Ca2+]i decreased upon application of the agonist and rapidly increased after restoration of calcium concentration to plasma level. We conclude that P1 and P2X receptors are present along the rabbit afferent arteriole and mediate calcium mobilization, with the same distribution in the proximal and distal segments. PMID- 10468654 TI - Nitric oxide and prostaglandin pathways interact in the regulation of hypercapnic cerebral vasodilatation. AB - To test whether nitric oxide and prostaglandin pathways interact in hypercapnic cerebral vasodilatation, cerebral blood flow (CBF) was measured in enflurane anaesthetized Sprague-Dawley rats using the hydrogen clearance method. Isometric tension was measured in rat middle cerebral arteries in vitro. The neuronal NO synthase inhibitor 7-nitroindazole (7-NI 60 mg kg-1 i.p.) reduced the hypercapnic CBF response by 62 +/- 7% (but not the hypoxic response) and indomethacin (IMC 6 mg kg-1 i.v.) reduced the hypercapnic CBF response by 60 +/- 5%. Combined application caused only an 80 +/- 1% reduction. The attenuation of hypercapnic CBF by IMC was diminished by 7-NI and similarly 7-NI had less effect in the presence of IMC. Spermine-NO (50 microM 0.5 microL min-1 intracortically) increased eucapnic and hypercapnic CBF in the presence of IMC. In isolated middle cerebral arteries, combined application of sodium nitroprusside (SNP 3 nM) and prostacyclin (30 nM) had a synergistic vasodilatory effect. Milrinone (PDE-III inhibitor) also potentiated prostacyclin-mediated vasodilatation. Our results suggest that the NO- and IMC-sensitive pathways involved in the hypercapnic response are distinct, however, both may interact synergistically. A similar synergism was observed between the effects of SNP and prostacyclin. PMID- 10468655 TI - Effects of [Na+]o, [Ca2+]o and cyclopiazonic acid on decline of post extrasystolic potentiation and twitch kinetics in guinea-pig and human myocardial preparations. AB - The decline of post-extrasystolic potentiation was investigated in atrial and ventricular muscle of guinea-pigs and in human atrial muscle. This decline is described in terms of the recirculation fraction (RF) for calcium in myocardial cells. Under control conditions, the mean values for RF were as follows: 0.61 +/- 0.03 in the guinea-pig atrium, 0.32 +/- 0.05 in the guinea-pig ventricle and 0.51 +/- 0.10 in the human atrium. Upon increasing calcium concentrations within the range of 0.5-10 mM or decreasing the sodium to 70%, the steady-state force and recirculation fraction increased concomitantly in all three types of muscles. Cyclopiazonic acid (CPA), in a 5-20 microM concentration, decreased force in the guinea-pig preparations and decreased RF. The effects of CPA were great in atrial muscle, 20 microM CPA decreasing RF from 0.61 to 0. 39. In ventricular muscle, this effect was small and not statistically significant. When changing calcium or sodium concentrations, increased force was accompanied by slower relaxation in atrial muscle, but an unchanged or slightly faster relaxation in ventricular muscle. In guinea-pig myocardium, CPA (5-20 microM) prolonged the time to peak force (TPF), but it slowed relaxation only in the ventricle. Reducing the calcium outflow by Na/Ca exchange increased RF as expected, and reducing the SR calcium re-uptake decreased RF. These interventions were also expected to reduce the rate of decline of intracellular calcium, but relaxation was not consistently prolonged. This indicates that factors other than the SR calcium pump and sarcolemmal Na/Ca exchange determine, or at least influence, the rate of relaxation. PMID- 10468656 TI - Ageing alters aortic antioxidant enzyme activities in Fischer-344 rats. AB - Oxidative stress imposed by reactive oxygen species is now believed to contribute to hypertension, atherosclerosis and ageing of the vasculature all involving a loss of relaxation. The antioxidant enzymes glutathione peroxidase, superoxide dismutase and catalase play a crucial role in defending against the ravages of oxidative stress. Our purpose was to characterize age-related changes in glutathione peroxidase, superoxide dismutase and catalase in the rat aorta. Aortas were extracted from seven young (4 months), seven middle aged (18 months) and seven old (24 months) animals. Analysis of variance was used with Fisher-LSD post hoc to determine mean differences among glutathione peroxidase, superoxide dismutase and catalase. Aortic glutathione peroxidase activities rose steadily with age expressed in micromol mg protein-1 min-1 +/- SEM (young: 141 +/- 22; middle aged: 198 +/- 18; old: 229 +/- 26) reaching significance between young and old. Superoxide dismutase activities significantly decreased in middle aged when compared with young (young: 22 +/- 2 vs. middle aged: 15 +/- 2 U mg protein-1) before trending upward again in old age (19 +/- 2). Catalase activities dropped significantly between young and old when expressed in mU mg protein-1 (young: 230 +/- 30; middle aged: 173 +/- 18; old: 144 +/- 23). Ratios for the various enzymes indicate a shrinking contribution of catalase with ageing, with an enhanced role for glutathione peroxidase in the antioxidant defence. These data in aortas of ageing rats show a complex alteration of the antioxidant profile. PMID- 10468657 TI - Changes in voltage activation of contraction in frog skeletal muscle fibres as a result of sarcoplasmic reticulum Ca2+-ATPase activity. AB - The effects of cyclopiazonic acid, a specific sarcoplasmic reticulum Ca2+-ATPase inhibitor, on isometric tension were studied in response to prolonged steady state depolarization induced by a rapid change in extracellular potassium concentration (potassium contractures) in frog semitendinosus muscle fibres. Cyclopiazonic acid (1-10 microM) enhanced the amplitude and time-course of relaxation of 146 mM potassium contracture. In the presence of cyclopiazonic acid 0.5 microM, the relationship between the amplitude of potassium contractures and the membrane potential shifted to more negative potentials, whereas the steady state inactivation curve was unchanged. These observations suggest that cyclopiazonic acid has no effect on voltage sensors. The difference between potassium contractures in the absence and presence of cyclopiazonic acid in skeletal muscle fibres implies that the amplitude and slow relaxation of tension during prolonged steady-state depolarization may be expected to depend not only on inactivation of the process regulating calcium release from the sarcoplasmic reticulum but also on the ability of the sarcoplasmic reticulum to pump calcium. PMID- 10468658 TI - Stimulation frequency-dependent reductions in skeletal muscle force and speed in creatine kinase-deficient mice. AB - Force and speed parameters were obtained from isometric contractions at different stimulation frequencies of creatine kinase-deficient and wildtype in situ mouse medial gastrocnemius muscles. The absence of creatine kinase did not affect force production at higher stimulation frequencies. However, at frequencies below 140 Hz, forces were lower than the controls (P < 0.05); at the lowest frequency applied (80 Hz) the force was reduced to approximately 60% compared with the wildtype muscles. In contrast, twitch force was not affected. When the contractions were preceded by a brief tetanus (50 ms), the effects of lacking creatine kinase on force production were more pronounced; at 80 Hz stimulation isometric force was further reduced to 66.5 +/- 6.2% (mean +/- SD; n=5) of the single contractions of the deficient muscles and to approximately 42% of the wildtype muscles. Twitch force was now also reduced (by approximately 50%) after the tetanus. The speed of the muscles was not affected in the single contractions. However, after a preceding tetanus, the rate of force rise was reduced by approximately 14% at high frequencies of stimulation. With decreasing frequencies (below 250 Hz), the reduction in speed became more pronounced; at 80 Hz the rate in the creatine kinase-deficient muscles was only 55.2 +/- 3.9% (mean +/- SD; n=5) of the wildtype muscles. No effects of the deficiency were found for the half relaxation times. The data suggest that an impaired creatine kinase system leads to lower activation levels at submaximal stimulation frequencies, possibly by a reduction in Ca2+-release during repetitive stimulation. Similar effects may be expected in normal fatigued muscle when phosphocreatine is depleted. PMID- 10468659 TI - Effects of oxytocin on the IGF-axis and some gastrointestinal hormones in ad libitum fed and food-restricted female rats. AB - The aims of this study were to investigate if administration of oxytocin to ad libitum fed and food-restricted female rats affects weight gain, body fatness, the IGF-axis, and some vagally mediated gastrointestinal hormones, such as gastrin, cholecystokinin (CCK) and somatostatin. Ad libitum fed and food restricted (receiving 70% of the food intake of the ad libitum fed group) female rats were injected subcutaneously, once a day, for 10 days, with saline (control) or oxytocin (1 mg kg-1 bodyweight). The animals were killed 5 days after the last injection. Oxytocin-treated food-restricted females had more body fat and lower plasma levels of IGF-I, IGFBP-1 and IGFBP-3 compared with saline-treated counterparts. Oxytocin-treated ad libitum fed rats also had lower plasma levels of IGFBP-1 but contained less body fat, compared with saline-treated counterparts. There was no effect of oxytocin treatment on body weight or weight gain in either of the feeding groups. Except for gastrin, which was lower, there was no effect of oxytocin on the gastrointestinal hormones studied. The results indicate that oxytocin treatment influences fat deposition and the IGF-axis in female rats, but that the results are dependent on the nutritional status of the animal. PMID- 10468660 TI - Physiological and morphological effects of perfusing isolated rat kidneys with hyperosmolal mannitol solutions. AB - Recently, we were able to modify the glomerular charge barrier using perfusates with low and normal ionic strengths keeping the osmolality unchanged. The concentration of fixed charges was reversibly reduced from 35 to 12 mEq L-1 as the solution with low content of NaCl was introduced with no apparent effect on the size selectivity. It can be argued however, that the mannitol used for maintenance of osmolality may induce changes in glomerular permeability per se. To explore this possibility, isolated kidneys were perfused at 8 degrees with hyperosmolal mannitol solutions (560 mOsm) and compared with those perfused with standard albumin solutions (295 mOsm). The vascular resistance (PRU100) fell from 0. 14 +/- 0.01 to 0.11 +/- 0.01 mmHg min 100 g mL-1 as the mannitol solution was introduced (P < 0.001). As the blood pressure should remain unchanged, the flow was increased from 8 to 11 mL min-1. The glomerular filtration rate (GFR) increased by 50% from 320 +/- 40 to 490 +/- 20 microL min-1 g-1 (P < 0.001). Despite these changes in haemodynamical parameters, there was no significant change in the fractional clearance for albumin. Kidneys perfused with the mannitol solution showed well-preserved histology, while there was a conspicuous collapse of the cortical tissue and signs of tubular epithelial swelling with the standard perfusate. Moreover, all glomeruli were perfused in the mannitol group, as revealed by fluorescence of FITC dextran, while the distribution was uneven in the control kidneys. We conclude that perfusion of isolated kidneys with a hyperosmolal mannitol solution increased GFR by increasing the number of functionally active nephrons with no apparent effect on the glomerular barrier, a pattern differing from alteration of ionic strength. PMID- 10468661 TI - Oxygen cost of dynamic leg exercise on a cycle ergometer: effects of gravity acceleration. AB - A model of the metabolic internal power (Eint) during cycling, which includes the gravity acceleration (ag) as a variable, is presented. This model predicts that Eint is minimal in microgravity (0 g; g=9. 81 m s-2), and increases linearly with ag, whence the hypothesis that the oxygen uptake (VO2) during cycling depends on ag. Repeated VO2 measurements during steady-state exercise at 50, 75 and 100 W on the cycle ergometer, performed in space (0 g) and on Earth (1 g) on two subjects, validated the model. VO2 was determined from the time course of decreasing O2 fraction during rebreathing. The gas volume during rebreathing was determined by the dilution principle, using an insoluble inert gas (SF6). Average VO2 for subject 1 at each power was 0.99, 1.21 and 1.52 L min-1 at 1 g (n=3) and 0.91, 1.13 and 1.32 L min-1 at 0 g (n=5). For subject 2 it was 0.90, 1.12 and 1. 42 L min-1 at 1 g, and 0.76, 0.98 and 1.21 L min-1 at 0 g. These values corresponded to those predicted from the model. Although resting VO2 was lower at 0 g than at 1 g, the net (total minus resting) exercise VO2 was still smaller at 0 g than at 1 g. This difference reflects the lower Eint at 0 g. PMID- 10468662 TI - Effects of recombinant erythropoietin (r-HuEPO) on plasma glucose concentration in endurance-trained rats. PMID- 10468663 TI - The impact of biochemical methods for single muscle fibre analysis. AB - Biochemical methods for single muscle fibre analysis provide sensitive measures for elucidating muscle fibre heterogeneity. The understanding of the complexity of skeletal muscle fibres, initially based on qualitative histochemistry and immunohistochemistry, has been greatly expanded by quantitative micromethods, such as microphotometry and microbiochemical assays. Assessment of metabolic enzyme activity levels has revealed pronounced scattering within and between different fibre types and has highlighted the use of specific enzyme activity ratios as discriminative measures. With the exception of type I fibres, metabolic properties are loosely coupled with molecular properties of the myofibrillar apparatus. As such, myosin heavy chain (MHC) isoforms appear to be the best choice for fibre type delineation. Among the two available methods for MHC-based fibre type distinction, single fibre electrophoresis appears to be superior to immunohistochemistry. The electrophoretic separation of MHC isoforms in single fibres is quantitative and, as opposed to immunohistochemistry, yields important information on MHC isoform proportions in hybrid fibres. Histochemical staining for myofibrillar ATPase activity can, thus, be correlated in most cases with specific MHC isoform profiles. Single fibre studies have demonstrated a relationship between ATP phosphorylation potential and MHC isoform complement. This relationship corresponds to different tension costs and provides an additional rationale for the MHC-based fibre type diversity and transitions. The combination of reverse transcriptase (RT) with polymerase chain reaction (PCR) has proved to be a highly sensitive tool and has extended single fibre analysis to the level of MHC mRNA isoforms. Application of RT-PCR techniques to single fibre fragments identified by their MHC protein isoform profile, provides insights at two levels of expression and, thus, has extended our knowledge on the plasticity of muscle and the dynamical state of muscle fibres. PMID- 10468664 TI - Cholinergic induced mesenteric vasorelaxation in response to head-up tilt. AB - Central hypovolaemia induced by head-up tilt evokes a reduction in superior mesenteric artery resistance resulting in maintenance of regional blood flow. Mechanisms of importance for this response are not known, but a parasympathetic contribution could be expected. To evaluate this hypothesis, superior mesenteric artery blood flow and resistance were evaluated by duplex ultrasound in eight healthy volunteers during postprandial head-up tilt with and without cholinergic blockade. During supine rest, cholinergic blockade did not influence the postprandial reduction in peripheral mesenteric artery resistance as expressed by analogous elevations in the diastolic blood velocity (to 62 +/- 9 vs. 56 +/- 7 cm s-1 with placebo). Throughout the normotensive and hypotensive phases of head-up tilt, cholinergic blockade reduced mesenteric artery mean blood velocity by 39 and 42%, respectively, corresponding to volume flow reductions by 35 and 41% (0.62 +/- 0.10 vs. 0.96 +/- 0.13 L min-1 and 0.52 +/- 0.07 vs. 0.88 +/- 0.16 L min-1; P < 0.05). Also, during both phases of head-up tilt, cholinergic blockade increased mesenteric artery resistance as reflected in a reduction in the diastolic blood velocity by 41 and 56%, respectively (44 +/- 4 vs. 74 +/- 13 cm s 1 and 24 +/- 6 vs. 54 +/- 8 cm s-1). These results support a cholinergic contribution to the mesenteric artery vasorelaxing response to central hypovolaemia induced by head-up tilt. PMID- 10468665 TI - Decreased responsiveness of vascular postjunctional alpha1-, alpha2-adrenoceptors and neuropeptide Y1 receptors in rats with heart failure. AB - Heart failure is associated with increased sympathetic nerve activity. We hypothesized that chronic sympathetic stimulation in heart failure resulted in decreased vascular sympathetic responsiveness. A pithed rat model was employed to evaluate peripheral vascular alpha-adrenoceptor and neuropeptide Y (NPY) receptor responsiveness. Heart failure was induced in Sprague-Dawley rats by coronary artery ligation. Sham operated rats (Sham) served as controls. Two months after this surgical procedure, both heart failure (n = 30) and Sham (n = 30) rats underwent standard pithing procedure. Pressor responses to preganglionic sympathetic nerve stimulation (PNS) and activation of postjunctional alpha1- and alpha2-adrenoceptors as well as Y1 receptors were studied. In response to PNS, cardiac index was similar between heart failure and sham rats (P = n.s.). Mean arterial pressure (MAP) increased in a frequency-dependent fashion after PNS in heart failure rats as well as in control rats. All the agonists used, i.e. the alpha1-adrenoceptor agonist phenylephrine, the alpha2-adrenoceptor agonists clonidine and BHT933 as well as NPY, induced dose-dependent increases in MAP in heart failure and in sham rats. However, in rats with heart failure, the response to all the agonists studied was significantly decreased and the dose response curves were shifted to the right (P < 0.01). We conclude that in vivo vascular response to postjunctional alpha1- and alpha2-adrenoceptors as well as Y1 receptors are decreased in rats with heart failure. PMID- 10468666 TI - Differentiation of the peptidergic vasoregulatory response to standardized splanchnic hypoperfusion by acute hypovolaemia or sepsis in anaesthetized pigs. AB - This study was performed to integratively investigate the vasoregulatory response during standardized splanchnic hypoperfusion in pigs. Splanchnic perfusion was reduced to 50% of baseline by: haemorrhage by 20 and 40% of the estimated total blood volume; femoral venous infusion of live E. coli to establish sepsis of systemic origin; portal venous infusion of live E. coli to establish sepsis of splanchnic origin. Invasive haemodynamic monitoring and radioimmunoassay analyses of arterial plasma concentrations of angiotensin II, endothelin-1 and atrial natriuretic peptide were carried out. Acute hypovolaemia reduced systemic and splanchnic vascular resistances following transient increases and increased angiotensin II levels (+587%), whereas endothelin-1 and atrial natriuretic peptide levels did not change significantly. Systemic sepsis following femoral venous infusion of E. coli resulted in increased splanchnic vascular resistance and increased levels of angiotensin II (+274%), endothelin-1 (+134%) and atrial natriuretic peptide (+185%). Infusion of E. coli via the portal venous route induced an increase in splanchnic vascular resistance associated with particularly elevated levels of angiotensin II (+1770%) as well as increased endothelin-1 (+201%) and atrial natriuretic peptide (+229%) concentrations. Hypovolaemia and sepsis, although standardized with a predefined level of splanchnic hypoperfusion, elicited differentiated cardiovascular and vasopeptidergic responses. Sepsis, particularly of portal origin, notably increased splanchnic vascular resistance related to increased production of the vasoconstrictors angiotensin II and endothelin-1. The role of atrial natriuretic peptide as a vasodilator seems to be of subordinate importance in hypovolaemia and sepsis. PMID- 10468667 TI - Increased metabolism of infused 1-methylxanthine by working muscle. AB - Exogenous substrates for capillary endothelial enzymes have potential as markers for changes in capillary recruitment (albeit nutritive flow). The metabolism of infused 1-methylxanthine (1-MX) to 1-methylurate (1-MU) by capillary endothelial xanthine oxidase of the constant-flow perfused rat hindlimb was shown previously to decrease with oxygen uptake (VO2) when nutritive flow was decreased. In the present study, the metabolism of 1-MX was investigated under conditions when VO2 and nutritive flow are known to increase during muscle contraction. The constant flow red blood cell-perfused rat hindlimb at 37 degrees C was used with sciatic nerve stimulation, and perfusate samples from whole hindlimb and working muscles taken for analysis of oxygen, lactate, 1-MX and 1-MU. Flow to muscle was assessed separately using fluorescent microspheres and was found to increase 2.3-fold to the working muscles while flow to the non-working leg muscles decreased to compensate. The activity of xanthine oxidase of whole muscle extracts was not altered by contraction. Samples from the vein draining the working muscles, and microsphere measurements of flow, indicated increased VO2 (5.5-fold to 249.2 +/- 43.1 micromol h-1 g-1, P < 0.001), and 1-MX conversion (2.5-fold to 1.87 +/- 0.25 micromol h-1 g-1, P < 0.01) (SEM are shown). It is concluded that as 1-MX metabolism parallels VO2, this substrate may be a useful indicator of changes in capillary (nutritive) surface area in muscle. PMID- 10468668 TI - Hyperoxia does not increase peak muscle oxygen uptake in small muscle group exercise. AB - We examined the influence of hyperoxia on peak oxygen uptake (VO2peak) and peripheral gas exchange during exercise with the quadriceps femoris muscle. Young, trained men (n=5) and women (n=3) performed single-leg knee-extension exercise at 70% and 100% of maximum while inspiring normal air (NOX) or 60% O2 (HiOX). Blood was sampled from the femoral vein of the exercising limb and from the contralateral artery. In comparison with NOX, hyperoxic arterial O2 tension (PaO2) increased from 13.5 +/- 0.3 (x +/- SE) to 41.6 +/- 0. 3 kPa, O2 saturation (SaO2) from 98 +/- 0.1 to 100 +/- 0.1%, and O2 concentration (CaO2) from 177 +/- 4 to 186 +/- 4 mL L-1 (all P < 0. 01). Peak exercise femoral venous PO2 (PvO2) was also higher in HiOX (3.68 +/- 0.06 vs. 3.39 +/- 0.7 kPa; P < 0.05), indicating a higher O2 diffusion driving pressure. HiOX femoral venous O2 saturation averaged 36.8 +/- 2.0% as opposed to 33.4 +/- 1.5% in NOX (P < 0.05) and O2 concentration 63 +/- 6 vs. 55 +/- 4 mL L-1 (P < 0.05). Peak exercise quadriceps blood flow (Qleg), measured by the thermo-dilution technique, was lower in HiOX than in NOX, 6.4 +/- 0. 5 vs. 7.3 +/- 0.9 L min-1 (P < 0.05); mean arterial blood pressure at inguinal height was similar in NOX and HiOX at 144 and 142 mmHg, respectively. O2 delivery to the limb (Qleq times CaO2) was not significantly different in HiOX and NOX. VO2peak of the exercising limb averaged 890 mL min-1 in NOX and 801 mL min-1 in HiOX (n.s.) corresponding to 365 and 330 mL min-1 per kg active muscle, respectively. The VO2peak-to-PvO2 ratio was lower (P < 0.05) in HiOX than in NOX suggesting a lower O2 conductance. We conclude that the similar VO2peak values despite higher O2 driving pressure in HiOX indicates a peripheral limitation for VO2peak. This may relate to saturation of the rate of O2 turnover in the mitochondria during exercise with a small muscle group but can also be caused by tissue diffusion limitation related to lower O2 conductance. PMID- 10468669 TI - Neuromuscular fatigue during repeated exhaustive submaximal static contractions of knee extensor muscles in endurance-trained, power-trained and untrained men. AB - The neural and muscular changes during fatigue produced in repeated submaximal static contractions of knee extensors were measured. Three groups of differently adapted male subjects (power-trained, endurance-trained and untrained, 15 in each) performed the exercise that consisted of 10 trials of submaximal static contractions at the level of 40% of maximal voluntary contraction (MVC) force till exhaustion with the inter-trial rest intervals of 1 min. MVC force, reaction time and patellar reflex time components before and after the fatiguing exercise and following 5, 10 and 15 min of recovery were recorded. Endurance-trained athletes had a significantly longer holding times for all the 10 trials compared with power-trained athletes and untrained subjects. However, no significant differences in static endurance between power-trained athletes and untrained subjects were noted. The fatigue test significantly prolonged the time between onset of electrical and mechanical activity (electromechanical delay) in voluntary and reflex contractions. The electromechanical delay in voluntary contraction condition for power-trained and untrained subjects and in reflex condition for endurance-trained subjects had not recovered 15 min after cessation of exercise. No significant changes in the central component of visual reaction time (premotor time of MVC) and latency of patellar reflex were noted after fatiguing static exercise. It is concluded, that in this type of exercise the fatigue development may be largely owing to muscle contractile failure. PMID- 10468670 TI - Total blood volume in endurance-trained postmenopausal females: relation to exercise mode and maximal aerobic capacity. AB - We have recently shown that postmenopausal female distance runners demonstrate elevated levels of blood volume compared with sedentary healthy peers. We also found a strong positive relation between blood volume and maximal oxygen consumption. In young adult males, endurance exercise training increases blood volume when performed in the upright, but not in the supine body position. Based on these observations, we hypothesized that among postmenopausal females, the elevation in blood volume would be absent or attenuated in women who train in the horizontal vs. upright body position, and that the lower blood volume in the former would be associated with lower maximal aerobic capacity. Thus, we measured supine resting plasma and total blood volumes (Evans blue dye) and maximal oxygen consumption in postmenopausal women: 10 sedentary controls, 10 swimmers and 10 runners matched for age (60 +/- 2; 59 +/- 2; 58 +/- 2 years, mean +/- SE) and hormone replacement use (5 per group). The swimmers and runners were further matched for training volume (4.5 +/- 0.2 vs. 4.8 +/- 0.6 h week-1), relative performance (78 +/- 5 vs. 75 +/- 3% of age-group world record) and fat-free mass (45.5 +/- 0. 8 vs. 44.9 +/- 1.5 kg). Total blood volume and maximal oxygen consumption were highest in the runners (81.2 +/- 4; 52.4 +/- 3 mL kg-1, respectively) and progressively lower in the swimmers (68.8 +/- 3; 44.2 +/- 2) and controls (59.2 +/- 2; 37.9 +/- 2; all P < 0. 05). In the pooled population, blood volume was positively related to maximal oxygen consumption (r = 0.72, P < 0.0001). We conclude that in endurance-trained postmenopausal females matched for training volume and competitive performance: (1) blood volume is lower in those who train in the horizontal (swimmers) compared with the upright position (runners); (2) the lower blood volume is associated with a lower maximal aerobic capacity. Nevertheless, blood volume and maximal oxygen consumption are higher in postmenopausal women who train in the horizontal position than in sedentary controls. PMID- 10468671 TI - Tachykinins stimulate acid and pepsinogen secretion in the isolated porcine stomach. AB - The precise role of tachykinins in regulation of acid and pepsinogen secretion has not been established. Tachykininergic effects on acid and pepsinogen secretion could be mediated either directly in the proximal stomach or through other indirect mechanisms, i.e. gastrin secretion. We studied the effects of the two tachykinins, substance P and neurokinin A, and of capsaicin, on acid and pepsinogen output, in isolated porcine non-antral stomach preparation. The release of substance P and neurokinin A was studied during electrical stimulation of the vagal nerves, and during capsaicin infusion. Substance P infusion (10-8 M) increased acid secretion from 30 +/- 8 to 68 +/- 17 fmol min-1 (n=6, P < 0.05) and pepsinogen output from 46 +/- 12 to 160 +/- 47 units of pepsin min-1 (n=9, P < 0.05). Neurokinin A also stimulated both acid and pepsinogen secretion, while capsaicin had no effect on either parameter. Electrical stimulation of the vagal nerves increased the release of both peptides. We conclude that tachykinins may be involved in regulation of acid and pepsinogen secretion. PMID- 10468672 TI - Review article: mechanisms of action and therapeutic applications of ursodeoxycholic acid in chronic liver diseases. AB - Ursodeoxycholic acid (ursodiol) is a non-toxic, hydrophilic bile acid used to treat predominantly cholestatic liver disorders. Better understanding of the cellular and molecular mechanisms of action of ursodeoxycholic acid has helped to elucidate its cytoprotective, anti-apoptotic, immunomodulatory and choleretic effects. Ursodeoxycholic acid prolongs survival in primary biliary cirrhosis and it improves biochemical parameters of cholestasis in various other cholestatic disorders including primary sclerosing cholangitis, intrahepatic cholestasis of pregnancy, cystic fibrosis and total parenteral nutrition-induced cholestasis. However, a positive effect on survival remains to be established in these diseases. Ursodeoxycholic acid is of unproven efficacy in non-cholestatic disorders such as acute rejection after liver transplantation, non-alcoholic steatohepatitis, alcoholic liver disease and chronic viral hepatitis. This review outlines the present knowledge of the modes of action of ursodeoxycholic acid, and presents data from clinical trials on its use in chronic liver diseases. PMID- 10468673 TI - Review article: malnutrition and maltreatment--a comment on orlistat for the treatment of obesity. AB - The prevalence of obesity has doubled in the last 10 years and is now reaching epidemic proportions. There is a significant comorbidity and financial cost associated with this disorder. Orlistat is an intestinal lipase inhibitor that is approved for the treatment of obesity. Recent randomized, double-blind, placebo controlled trials have demonstrated the benefit of orlistat used in conjunction with a hypocaloric (low-fat) diet in facilitating weight reduction and the long term maintenance of this weight loss. Patients treated with orlistat lost a greater amount of initial body weight compared to those who received placebo. After 24 months of treatment, weight loss of more than 5% was maintained in a greater number of those treated with orlistat. This was associated with significant reductions in cardiovascular risk factors (cholesterol, LDL cholesterol, LDL:HDL cholesterol ratio). The main adverse events are related to fat malabsorption, with potential losses of fat-soluble vitamins and other compounds. Orlistat as a treatment for obesity, when prescribed within present guidelines, can aid modest weight loss in about one-third of patients. More importantly, it can assist in the maintenance of weight loss with major medical benefits for these patients. PMID- 10468674 TI - Economic analysis of step-wise treatment of gastro-oesophageal reflux disease. AB - BACKGROUND: To expose patients with gastro-oesophageal reflux disease (GERD) to the least amount of medication and to reduce health expenditures, it is recommended that their treatment is started with a small dose of an antisecretory or prokinetic medication. If patients fail to respond, the dose is increased in several consecutive steps or the initial regimen is changed to a more potent medication until the patients become asymptomatic. Although such treatment strategy is widely recommended, its impact on health expenditures has not been evaluated. METHODS: The economic analysis compares the medication costs of competing medical treatment strategies, using two different sets of cost data. Medication costs are estimated from the average wholesale prices (AWP) and from the lowest discount prices charged to governmental health institutions. A decision tree is used to model the step-wise treatment of GERD. In a Monte Carlo simulation, all transition probabilities built into the model are varied over a wide range. A threshold analysis evaluates the relationship between the cost of an individual medication and its therapeutic success rate. RESULTS: In a governmental health care system, a step-wise strategy saves on average $916 per patient every 5 years (range: $443-$1628) in comparison with a strategy utilizing only the most potent medication. In a cost environment relying on AWP, the average savings amount to $256 (-$206 to +$1561). The smaller the cost difference between two consecutive treatment steps, the longer one needs to follow the patients to reap the benefit of the small cost difference. However, even a small cost difference can turn into tangible cost savings, if a large enough fraction of GERD patients responds to the initial step of a less potent but also less expensive medication. CONCLUSIONS: The economic analysis suggests that a step wise utilization of increasingly more potent and more expensive medications to treat GERD would result in appreciable cost savings. PMID- 10468675 TI - The effect of omeprazole on gastro-oesophageal reflux and symptoms during strenuous exercise. AB - BACKGROUND: Strenuous exercise exacerbates gastro-oesophageal reflux and symptoms and this may be diminished by antisecretory medication with omeprazole. METHODS: Fourteen well-trained athletes (13 men, one woman), who indicated suffering from either heartburn, regurgitation or chest pain during competition running, performed two experimental trials at 2-week intervals using a randomized, double blind, placebo-controlled crossover design. During the 6 days preceding the trial and on the trial day itself either 20 mg of omeprazole or a placebo was administered. Two hours after a low-fat breakfast and 1 h after the last study dose, the trial started with five successive 50-min periods: rest, three running periods on a treadmill, and recovery. Reflux (percentage time and number of periods oesophageal pH <4) was measured with an ambulant pH system during these periods. RESULTS: Compared to rest, reflux lasted significantly longer and occurred more frequently during the first running period, irrespective of the intervention, whereas during the second running period this effect was only observed with the placebo. Reflux occurred for longer and more frequently with the placebo than with omeprazole, but this was significant during the first two running periods only. Seven subjects reported heartburn, regurgitation and/or chest pain during exercise, irrespective of the intervention. Only a minority of the symptom periods was actually associated with acid reflux and in all cases this concerned periods with heartburn. CONCLUSIONS: Running-induced acid reflux, but not symptoms, were decreased by omeprazole, probably because most symptoms were not related to acid reflux. PMID- 10468676 TI - Safety and efficacy of pantoprazole 40 mg daily as relapse prophylaxis in patients with healed reflux oesophagitis-a 2-year follow-up. AB - BACKGROUND: Pantoprazole is a benzimidazole derivative which selectively inhibits the proton pump H+, K+-ATPase, necessary for the final step in gastric acid secretion. AIM: To assess safety and efficacy of oral pantoprazole (40 mg o.d.) used as a prophylaxis against relapse in patients with healed reflux oesophagitis during an open-label, 2-year study. METHODS: Outpatients (n=157) with healed stage II or III reflux oesophagitis (Savary-Miller classification) were enrolled into a long-term, multicentre maintenance study. Endoscopy was performed at entry into the study, after 12 and 24 months, or when disease-specific symptoms occurred on more than three consecutive days. Symptoms were assessed at 3-monthly intervals. Endoscopically confirmed relapses (at least stage I) were evaluated as treatment failures. RESULTS: Of the 178 adverse events, experienced by 88 (56%) patients (intention-to-treat population), 12 (7%) were assessed by the investigators as possibly related to the study medication. Median serum gastrin levels increased from a baseline of 46 ng/L to 90 ng/L, reaching a plateau after 9 months. For the intention-to-treat population the endoscopic remission rates after 12 and 24 months were 87% and 76%, respectively (Life-Table survival analysis, Kaplan-Meier). CONCLUSION: Pantoprazole 40 mg proved to be safe and efficacious during a 2-year prophylaxis treatment in patients with healed reflux oesophagitis. PMID- 10468677 TI - Patterns of consultation and treatment for heartburn: findings from a Singaporean community survey. AB - BACKGROUND: The prevalence of reflux-type symptoms among community individuals in Singapore is low. AIM: To describe the healthcare-seeking behaviour of those subjects with heartburn. METHODS: A cross-sectional survey, using a reliable and valid questionnaire, was carried out on a race-stratified random sample of residents, aged 21-95 years, in a Singaporean town; 93% responded (n = 696). RESULTS: The ethnic-adjusted prevalence of heartburn for the past 1-year was 4.6%. Sixteen (30%) of the 53 heartburn sufferers had sought consultation for their symptoms. They were more likely to have severe heartburn (P<0.001), and to have complained of nocturnal awakening due to heartburn (P<0.05) than those who did not present to medical attention. Ethnic origin did not influence the consultation pattern. Only 18 (34%) of the 53 heartburn sufferers received pharmacological therapy for their symptoms. The most commonly used medication was antacid. Medication use was associated with symptom severity (P<0.05), but not ethnicity. CONCLUSIONS: Heartburn is uncommon in the general population of Singapore. Few heartburn sufferers seek medical attention, and most do not receive medications for symptomatic control. The decision to seek medical advice and/or to medicate was generally linked to symptom severity, but not to ethnicity. PMID- 10468678 TI - On demand therapy of reflux oesophagitis--a prospective study of symptoms, patient satisfaction and quality of life. AB - BACKGROUND: In patients with low-grade reflux oesophagitis adequate symptom control is the aim of treatment. Effervescent tablets alleviate heartburn more rapidly than ordinary tablets. AIM: To investigate symptom control, patient satisfaction, health-related quality of life and disease progress when ranitidine 150 mg effervescent tablets were offered as on demand treatment. We also wanted to investigate whether any biological or psycho-social factor could predict patient satisfaction. METHOD: Consecutive patients with endoscopically verified reflux oesophagitis grade I-II were followed up for 12 months. 24 h pH-metry, disease history, symptoms and several psycho-social factors were registered at baseline and 12 months follow-up. RESULTS: Eighty-one patients were included. Mean age was 50.7 years (range 21-82), 63% were men. Mean tablet consumption was 1.21 per day (range 7-1016 tablets/year). At the 1-year follow-up discomfort resulting from reflux symptoms was significantly reduced (P<0.001), and the patients' social and vocational life improved. Eighty-four percentage of the patients were satisfied with the treatment. 24 h pH-metry or number of reflux episodes did not change. We did not find any factors able to predict patient satisfaction. CONCLUSIONS: On demand therapy with ranitidine effervescent tablets was well accepted by the majority of patients with reflux oesophagitis grade I. Even though the number of reflux episodes did not change, the patients experienced less discomfort due to reflux symptoms, and their social and vocational life was better. There was no significant progression of the disorder during the 1-year follow-up. No predictive factor for patient satisfaction was found. PMID- 10468679 TI - Omeprazole therapy decreases the need for dilatation of peptic oesophageal strictures. AB - BACKGROUND: Better control of gastric acid secretion with omeprazole appeared to decrease the need for dilatation of oesophageal strictures complicating gastro oesophageal reflux disease in our hospital-based endoscopy service. AIM: To investigate whether the perceived decrease in the need for oesophageal dilatation could be documented from endoscopy records, and, if confirmed, whether this could be related to the treatment used. PATIENTS AND METHODS: Retrospective study of the records of 69 patients who had peptic oesophageal strictures dilated, followed by treatment with acid inhibition for at least 6 months. Mean duration of follow-up was 3.9 years during treatment with H2-receptor antagonists and 2.1 years while on omeprazole (258 and 78 patient-years, respectively). Re-dilatation rates were compared between those treated with H2-receptor antagonists or omeprazole. RESULTS: There has been a significant decrease in dilatations performed for gastro-oesophageal reflux induced strictures (P<0.001), while dilatation rates for other indications remained constant. Treatment with omeprazole not only decreased the need for further dilatations, but also prolonged the mean time between any further dilatations to 26.3 months compared to 9.3 months for those on an H2-receptor antagonist (P<0.0001). CONCLUSIONS: Following dilatation of peptic oesophageal strictures, treatment with omeprazole in place of an H2-blocker significantly decreases the need for repeat dilatation. PMID- 10468680 TI - A systematic review of Helicobacter pylori eradication therapy--the impact of antimicrobial resistance on eradication rates. AB - BACKGROUND: We systematically reviewed all available data in the literature to determine the overall eradication rates of currently advised Helicobacter pylori eradication regimens and to resolve conflicting evidence on the impact of antimicrobial resistance on the eradication rates. METHODS: A comprehensive search of all published trials on H. pylori eradication therapy was carried out via an electronic database search, hand-searching and checking reference lists of pharmaceutical companies and other reviews. Full papers and abstracts in the English language which study currently advised eradication regimes were included. RESULTS: 770 study-arms were analysed. Mean eradication rates for bismuth based triple, proton pump inhibitor triple, quadruple and ranitidine bismuth citrate combination therapies vary from 65 to 92%. In case of nitroimidazole resistance, a drop in efficacy of up to 50% was found for bismuth-based triple and proton pump inhibitor-based triple therapies. For quadruple therapy, a significant difference in efficacy was found in the equal-effects analysis; however, this could not be confirmed in the random-effects analysis. In case of clarithromycin resistance, a mean drop in efficacy of 56% was found for one- and two-week clarithromycin containing proton pump inhibitor-triple therapies and of 58% for two-week ranitidine bismuth citrate combined with clarithromycin therapies. For ranitidine bismuth citrate combined with clarithromycin and nitroimidazole, no difference in efficacy was found in case of nitroimidazole or clarithromycin resistance, but data are still scarce. CONCLUSIONS: The cure rate with most regimens dropped significantly, in case of nitroimidazole-resistant strains, compared to nitroimidazole-susceptible strains. In case of clarithromycin resistance, the efficacy of most regimens is also decreased; however, data are still scarce. These data should allow physicians to make a better choice of an appropriate therapy for their patients. PMID- 10468681 TI - Citric acid or orange juice for the 13C-urea breath test: the impact of pH and gastric emptying. AB - BACKGROUND: There is an ongoing debate about the optimal test drink to be used in the 13C-urea breath test (13C-UBT). We recently reported that a citric acid solution is the optimal test drink in the 13C-UBT, because it provides a high 13CO2 recovery and the excellent accuracy of the test appears optimal compared to other test meals. Orange juice, because of a better taste, is also propagated as a test drink in the 13C-UBT. AIM: To compare the diagnostic accuracy of the 13C UBT with either orange juice or citric acid solution as a test drink. Furthermore, the effect of these test drinks on the gastric emptying rate was determined. METHODS: H. pylori status was assessed by histology, rapid urease test and culture in 50 consecutive dyspeptic patients. A 13C-UBT was performed on two consecutive days by giving 75 mg of 13C-urea randomly dissolved in 200 mL 0.1 M citric acid solution or 200 mL orange juice. The 13CO2/12CO2 ratio was measured in breath samples taken before and 15, 30, 45 and 60 min after administration of the test drink. The gastric emptying rate of orange juice and citric acid solution was compared to that of water in 10 healthy subjects on three consecutive days by means of a 13C-sodium acetate breath test; 50 mg of 13C sodium acetate was dissolved in 200 mL of each solution and breath samples were collected before and every 10 min for 90 min after administration of the test drink. RESULTS: Twenty-six out of 50 patients (52%) were infected with H. pylori. Significantly higher values over baseline (35.7+/-5.2 per thousand vs. 23.2+/-3.4 per thousand, P<0.001) and higher area under the curve (1507+/-198 vs. 927+/-128, P<0.001) were observed in H. pylori-positive patients when citric acid solution was administered compared with orange juice. Sensitivity of the 13C-UBT was 100% when citric acid was used as a test drink and 88% with orange juice. Specificity was 100% with both test drinks. Gastric emptying of citric acid solution (t1/2 = 60.9+/-3.5 min) was significantly slower than that of orange juice (t1/2 = 49.7+/ 3.1 min, P<0.001). CONCLUSION: 13C-UBT loses diagnostic accuracy when orange juice instead of citric acid is used as a test drink. The faster gastric emptying of orange juice might be responsible for the lower diagnostic accuracy of the 13C UBT. PMID- 10468682 TI - Famotidine versus omeprazole in combination with clarithromycin and metronidazole for eradication of Helicobacter pylori--a randomized, controlled trial. AB - BACKGROUND: One-week low-dose triple therapy is currently considered the gold standard regimen for treatment of Helicobacter pylori infection. However, the mechanisms involved in the synergy between antibiotics and proton pump inhibitors are controversial. AIMS: To test the hypothesis that acid suppression represents the crucial mechanism by which the antibacterial activity of antibiotics can be enhanced, and to assess the impact of primary resistance on treatment outcome. METHODS: One hundred and twenty patients with H. pylori infection and duodenal ulcer, gastric ulcer or non-ulcer dyspepsia were randomly assigned to a 1 week course of either famotidine 80 mg b.d., clarithromycin 250 mg b.d. and metronidazole 500 mg b.d. (FCM group; n = 60) or omeprazole 20 mg o.d., clarithromycin 250 mg b.d. and metronidazole 500 mg b.d. (OCM group; n = 60). Gastroscopy was performed at baseline and 5 weeks after completion of treatment. H. pylori status was assessed by biopsy urease test, histology and culture. RESULTS: In the intention-to-treat analysis, eradication of H. pylori was achieved in 47 of 60 patients (78%; 95% CI: 66-88%) in the FCM group, compared to 44 of 60 patients (73%; 95% CI: 60-84%) in the OCM group (N.S.). Using per protocol analysis, eradication therapy was successful in 47 of 52 patients (90%; 95% CI: 79-97%) treated with FCM and 44 of 57 patients (77%; 95% CI: 64-87%) treated with OCM (N.S.). Primary metronidazole resistance was present in 27% and primary clarithromycin resistance in 8% of strains. Overall per protocol eradication rates in strains susceptible to both antibiotics and strains with isolated metronidazole resistance were 93% and 84%, respectively. No patient with clarithromycin resistance responded to treatment. CONCLUSIONS: High-dose famotidine and omeprazole, combined with clarithromycin and metronidazole, are equally effective for eradication of H. pylori. In 1-week low-dose triple therapy, metronidazole resistance has no major impact on eradication rates whereas clarithromycin resistance is associated with a poor treatment outcome. PMID- 10468683 TI - Comparison of ranitidine bismuth citrate plus clarithromycin with omeprazole plus clarithromycin for the eradication of Helicobacter pylori. AB - BACKGROUND: Many dual and triple therapy treatment regimens have been proposed for the eradication of Helicobacter pylori. However, assessing the relative efficacy of these regimens is complicated by differences in study design, and few well-controlled comparative studies have been reported. METHODS: This multicentre, randomized, double-blind study involved 530 duodenal ulcer patients, of whom 520 had confirmed H. pylori infection. Patients received 14 days b.d. dual therapy of either ranitidine bismuth citrate (RBC) 400 mg or omeprazole 20 mg, both with clarithromycin 500 mg to eradicate H. pylori, followed by a further 14 days of treatment with RBC 400 mg b. d. or omeprazole 20 mg o.d. to facilitate ulcer healing. H. pylori eradication and ulcer healing were assessed at least 26 days after the end of treatment. Adverse events were recorded throughout the study. RESULTS: H. pylori was eradicated in 90% of patients who received RBC with clarithromycin and in 66% of patients who received omeprazole with clarithromycin (per protocol; P<0.001). intention-to-treat eradication rates were 77% and 60%, respectively (P<0.001). Ulcer healing rates were 97% in the RBC treatment group and 95% in the omeprazole treatment group. Only 3% and 1% of patients in the RBC and omeprazole treatment groups, respectively, were withdrawn due to adverse events. CONCLUSIONS: RBC with clarithromycin is a simple and highly effective dual therapy regimen for the eradication of H. pylori, and is significantly more effective than omeprazole with clarithromycin. Both treatment regimens are well tolerated and effectively heal duodenal ulcers. PMID- 10468684 TI - One-week ranitidine bismuth citrate in combinations with metronidazole, amoxycillin and clarithromycin in the treatment of Helicobacter pylori infection: the RBC-MACH study. AB - BACKGROUND: We have previously shown that ranitidine bismuth citrate (RBC)-based triple therapy is comparable to proton pump inhibitor-based triple therapy in eradicating Helicobacter pylori infection. AIM: To test the efficacy of different combinations of antimicrobials with RBC in the treatment of H. pylori infection. METHODS: Dyspeptic patients with H. pylori infection were prospectively randomized to receive one of the following regimens: (i) RBC 400 mg, amoxycillin 1 g, clarithromycin 500 mg [RAC]; (ii) RBC 400 mg, metronidazole 400 mg, clarithromycin 500 mg [RMC]; (iii) RBC 400 mg, metronidazole 400 mg, tetracycline 1 g [RMT] (all given twice daily for 1 week); or (iv) RBC 400 mg plus clarithromycin 500 mg twice daily for 2 weeks [RC-2]. Endoscopy (rapid urease test and culture) and 13C-urea breath test (UBT) were performed before randomization. Four weeks after finishing medication, the 13C-UBT was repeated in all cases and endoscopy was offered to patients with peptic ulcers. RESULTS: Four hundred patients were randomized but in two (one in the RAC group and one in the RMC group) H. pylori infection was not confirmed. Successful eradication of H. pylori (intention-to-treat analysis and 95% CI) of RAC (86% [79-93%]), RMC (90% [84-96%]), RMT (79% [71-87%]) and RC-2 (82% [75-90%]) were comparable, with a trend favouring clarithromycin-containing triple therapy regimens. Among 276 isolates tested for antibiotic sensitivity, primary resistance to metronidazole, clarithromycin and amoxycillin was found in 56%, 2% and 0.4%, respectively. When given RMC or RMT, patients infected by metronidazole-resistant H. pylori had success in eradicating H. pylori similar to patients infected by metronidazole sensitive H. pylori. CONCLUSION: One-week RBC triple therapy is effective in curing H. pylori infection. PMID- 10468685 TI - The safety of proton pump inhibitors in pregnancy. AB - AIM: To assess the safety of proton pump inhibitors during pregnancy. METHODS: Fifty-one pregnant women exposed to proton pump inhibitors around the time of conception or during pregnancy were compared with 13 327 controls without exposure to any prescribed drug in a population-based study based on The Pharmaco Epidemiological Prescription Database of North Jutland and the Danish Hospital Discharge Registry. RESULTS: Three babies with malformations were found among 38 women exposed to proton pump inhibitors from 30 days before conception to the end of the first trimester. No cases of stillbirth were recorded. Crude relative risks of malformation, low birth weight and preterm delivery were 1.6 (95% CI: 0.5-5.1), 1.8 (95% CI: 0.2-13.0) and 2.3 (95% CI: 0.9-6.0), respectively. CONCLUSIONS: In this population-based follow-up study, we found no substantially elevated risk in terms of malformations, low birth weight or number of preterm deliveries in pregnancies exposed to proton pump inhibitors. However, further monitoring is warranted in order to establish or rule out a potential association between the use of proton pump inhibitors and increased risk of either cardiac malformations or preterm birth. PMID- 10468686 TI - The effects on intragastric acidity of per-gastrostomy administration of an alkaline suspension of omeprazole. AB - BACKGROUND: It may be difficult to administer proton pump inhibitors via gastrostomy. Previous studies have examined the effect of intact proton pump inhibitor granules in orange juice. This study examined the effect of an alkaline suspension of omeprazole (simplified omeprazole suspension (SOS)) on 24-h intragastric acidity. METHODS: Six men with an established gastrostomy had a baseline 24-h intragastric pH study using methodology we have previously described. They then received 20 mg SOS o.d. for 7 days and had a repeat pH study at the end of this period. Four of the patients then received 20 mg SOS with 30 cc of liquid antacid (Mylanta) per gastrostomy o.d. for a further 7 days and then underwent a third pH study. RESULTS: SOS raised mean pH from 2.2 to 4.1. Intragastric pH was above 3, 4 and 5 for 35, 28 and 17% of the 24-h period at baseline, respectively; corresponding values after SOS were 63, 51 and 39%, respectively. Addition of liquid antacid to SOS did not further increase its pH controlling effect. CONCLUSIONS: We found a statistically significant effect of o.d. SOS on intragastric pH when administered via gastrostomy. We found no additional benefit of administering SOS with liquid antacid. PMID- 10468687 TI - One-year follow-up of newly diagnosed irritable bowel syndrome patients. AB - BACKGROUND: Irritable bowel syndrome is the most common functional gastrointestinal disorder seen by general physicians. METHODS: We followed up a population-based cohort of newly diagnosed irritable bowel syndrome patients aged 20-79 years, to examine patterns of treatment, comorbidity and healthcare utilization. We used the UK General Practice Research Database as the source population. Individuals with other gastrointestinal diseases, cancer and pregnant women were not included. There were 2956 patients in our final cohort. RESULTS: Irritable bowel syndrome patients were mainly young and middle-aged; only 12% were 60 years or older. The majority of patients were women (74%). There were no marked differences in terms of use of healthcare services or comorbidity status in the year before irritable bowel syndrome diagnosis as compared to the year after. Fourteen per cent of irritable bowel syndrome patients received no drug treatment at all. Among those treated, the first choice was an antispasmodic. Elderly patients (>60 years old) were more likely to receive drug treatment. Females had a slightly higher probability of being treated than men, except for the category of anti-diarrhoeal drugs. CONCLUSION: This study has shown that irritable bowel syndrome patients are mainly young and female. We also found that treatment pattern varied according to age and gender. Elderly patients and females were at a higher risk of receiving drug treatment. PMID- 10468688 TI - Impact on the composition of the faecal flora by a new probiotic preparation: preliminary data on maintenance treatment of patients with ulcerative colitis. AB - BACKGROUND: Although 5-aminosalicylic acid (5-ASA) oral compounds are the standard maintenance treatment for ulcerative colitis in remission, some patients cannot use them because of side-effects. Clinical and experimental observations have suggested the potential role of probiotics in inflammatory bowel disease therapy. AIM: To evaluate the effects on intestinal microflora and the clinical efficacy of a new probiotic preparation in patients with ulcerative colitis in remission. PATIENTS AND METHODS: Twenty patients with ulcerative colitis, intolerant or allergic to 5-ASA, have been treated with a new probiotic preparation (VSL#3, CSL, Milan, Italy) containing 5x10(11) cells/g of 3 strains of bifidobacteria, 4 strains of lactobacilli and 1 strain of Streptococcus salivarius ssp. thermophilus. Two doses of 3 g were administered o.d. for 12 months. Faecal samples for stool culture were obtained from the patients at the beginning of the trial and after 10, 20, 40, 60, 75, 90 days, 12 months and at 15 days after the end of the treatment. The following bacterial groups have been evaluated in the faeces: total aerobic and anaerobic bacteria, enterococci, Streptococcus thermophilus, lactobacilli, bifidobacteria, Bacteroides, clostridia, coliforms. Patients were assessed clinically every two months, and assessed endoscopically at 6 and 12 months or in relapse. RESULTS: Faecal concentrations of Streptococcus salivarius ssp. thermophilus, lactobacilli and bifidobacteria increased significantly in all patients, compared to their basal level, from the 20th day of treatment (P<0.05) and remained stable throughout the study. Concentrations of Bacteroides, clostridia, coliforms, total aerobic and anaerobic bacteria did not change significantly during treatment (P = N.S.). Fifteen of 20 treated patients remained in remission during the study, one patient was lost to follow up, while the remaining relapsed. No significant side effects have been reported. CONCLUSIONS: These results show that this probiotic preparation is able to colonize the intestine, and suggest that it may be useful in maintaining the remission in ulcerative colitis patients intolerant or allergic to 5-ASA. Controlled trials are warranted to confirm these preliminary results. PMID- 10468689 TI - Enteric coating of aspirin significantly decreases gastroduodenal mucosal lesions. AB - BACKGROUND: Low-dose aspirin (acetylsalicylic acid, ASA) increases the risk of developing peptic ulceration. AIM: To investigate the gastroduodenal mucosal tolerability of enteric-coated ASA (EC-ASA) 100 mg/day compared to either placebo (study 1) or plain ASA 100 mg/day (study 2) in healthy volunteers. METHODS: Study 1: In this double-blind study 18 volunteers received randomized dosing with either EC-ASA 100 mg or placebo for 15 days. Study 2: 41 volunteers underwent randomized 7-day dosing of either EC-ASA 100 mg or plain ASA 100 mg in this double-blind, parallel-group, comparison study. In both studies acute gastroduodenal mucosal lesions were assessed endoscopically before treatment, on the morning of day 1 after the first dose (only in study 2), and on the morning after the last dose of the test medication. RESULTS: Study 1 did not reveal any significant differences between the lesion scores of EC-ASA and placebo. In contrast, in study 2 significantly higher total gastroduodenal mucosal lesion scores were observed on day 1 after the first dose and after 7 days of dosing with plain ASA (mean sum of the lesion scores in the gastric fundus, body, antrum and in the duodenal bulb: day 1: plain ASA 3.95+/-3.38 vs. EC-ASA 1.43+/-1.91, P = 0.03; day 7: plain ASA 6.35+/-4.10 vs. EC-ASA 2.00+/-2.02, P = 0.0004). Tolerance of the test drugs was good, and no other adverse events were observed. CONCLUSIONS: Enteric-coated aspirin 100 mg/day causes significantly less gastroduodenal damage over 7 days than the same dose of plain aspirin, when given to healthy subjects. There was little gastric injury and no significant differences between EC-ASA and placebo in this respect. PMID- 10468690 TI - Selective COX-2 inhibitors and human inflammatory bowel disease. AB - BACKGROUND: Much recent effort has been made to produce selective inhibitors of cyclo-oxygenase-2 (COX-2) in the belief that these will lack the gastrointestinal damaging effects of traditional non-steroidal anti-inflammatory drugs (NSAIDs). Inflammatory bowel disease is associated with increased local production of prostanoids. These prostanoids, particularly PGE2 and PGI2, may well be protective as inflammatory bowel disease is aggravated by NSAID use. AIM: To examine the effects of a traditional NSAID and a highly selective COX-2 inhibitor on the production of these prostanoids in human inflammatory bowel disease. METHODS: Colonic mucosal biopsies were obtained from patients undergoing routine colonoscopy and biopsy for diagnostic or surveillance purposes. Biopsies were incubated in culture medium containing 10% foetal calf serum and antibiotics, plus test drugs or vehicle for 24 h, after which time the medium was removed and the content of PGE2, PGI2 (measured as 6 keto-PGF1alpha) and thromboxane (Tx) A2 (measured as TxB2) determined. RESULTS: Biopsies obtained from diseased colonic mucosa produced significantly more PGE2, PGI2 and thromboxane A2 than did controls (for example, PGE2: ulcerative colitis, 4.17+/-1.06; Crohn's disease, 3.97+/-1.66; control, 0.12 +/-0.13 ng/mL, n = 8-12). These increases were inhibited to a similar extent by either a highly selective COX-2 inhibitor (L 745,337) or a traditional non-selective NSAID (indomethacin). CONCLUSIONS: Until selective COX-2 inhibitors have been assessed adequately in human inflammatory bowel disease, these compounds should not be assumed to be safe for the gastrointestinal tract in inflammatory bowel disease. PMID- 10468691 TI - Long-term haemodynamic effects of octreotide on postprandial splanchnic hyperemia in humans: a placebo-controlled echo-doppler study. AB - BACKGROUND: Octreotide is a potent splanchnic hypotensive somatostatin analogue effective in the treatment of acute variceal bleeding. AIM: To study the effects of octreotide on basal and postprandial splanchnic and systemic haemodynamics, and hormonal changes in humans. METHODS: Twenty-four healthy volunteers were randomized to receive a liquid meal and either octreotide (OCT, 100 microg bolus) or placebo repeatedly every 4 h for 48 h. Splanchnic (Doppler ultrasound) and systemic haemodynamics (non-invasive cardiac monitoring) were assessed for 2 h on four consecutive days: one control day and after doses 1 (0 h), 7 (24 h) and 13 (48 h). RESULTS: The maximum postprandial increases in mean blood velocity of the superior mesenteric artery (SMA-Vmean +72%), portal (PBF +52%) and total hepatic blood flow (HBF +50%) observed in the placebo group, were abolished after the first dose of octreotide (SMA-Vmean -23%, P<0.01; PBF -22%, P<0.01; HBF -21%, P<0.01). Postprandial hyperemia was restored at the end of the 48-h study period, but baseline SMA-Vmean (placebo 40+/-12, OCT 29+/-11 cm/s, P<0.05) and PBF (placebo 1200+/-971, OCT 743+/-449 mL/min, P<0.05) remained significantly lower in the octreotide group. The postprandial decrease of systemic vascular resistance and increase of cardiac index were prevented by octreotide for 48 h. CONCLUSIONS: Repeated 4-hourly bolus injections of octreotide reduce splanchnic blood flow for at least 48 h, but the prevention of food-induced splanchnic hyperemia is short-lasting. PMID- 10468692 TI - Neutrophils in NSAID-induced injury. PMID- 10468695 TI - Management of uninvestigated and functional dyspepsia: a Working Party report for the World Congresses of Gastroenterology 1998. AB - BACKGROUND: The management of dyspepsia is controversial. METHODS: An international Working Party was convened in 1998 to review management strategies for dyspepsia and functional dyspepsia, based on a review of the literature and best clinical practice. RESULTS: Dyspepsia, defined as pain or discomfort centred in the upper abdomen, can be managed with reassurance and over-the-counter therapy if its duration is less than 4 weeks on initial presentation. For patients with chronic symptoms, clinical evaluation depends on alarm features including patient age. The age cut off selected should depend on the age specific incidence when gastric cancer begins to increase, but in Western nations 50 years is generally an acceptable age threshold. In younger patients without alarm features, Helicobacter pylori test and treatment is the approach recommended because of its value in eliminating the peptic ulcer disease diathesis. If, after eradication of H. pylori, symptoms either are not relieved or rapidly recur, then an empirical trial of therapy is recommended. Similarly, in H. pylori-negative patients without alarm features, an empirical trial (with antisecretory or prokinetic therapy depending on the predominant symptom) for up to 8 weeks is recommended. If drugs fail, endoscopy should be considered because of its reassurance value although the yield will be low. In older patients or those with alarm features, prompt endoscopy is recommended. If endoscopy is non-diagnostic, gastric biopsies are recommended to document H. pylori status unless already known. While treatment of H. pylori is unlikely to relieve the symptoms of functional dyspepsia, the long-term benefits probably outweigh the risks and treatment can be considered on a case-by-case basis. In H. pylori-negative patients with documented functional dyspepsia, antisecretory or prokinetic therapy, depending on the predominant symptom, is reasonable, assuming reassurance and explanation are insufficient, unless patients have already failed this approach. Other treatment options include antidepressants, antispasmodics, visceral analgesics such as serotonin type 3 receptor antagonists, and behavioural or psychotherapy although these are all of uncertain efficacy. Long term drug treatment in functional dyspepsia should be avoided; intermittent short courses of treatment as needed is preferred. CONCLUSION: The management of dyspepsia recommended is based on current best evidence but must be tailored to local factors such as practice setting, the background prevalence of H. pylori and structural disease, and costs. PMID- 10468696 TI - Improvement in pain and bowel function in female irritable bowel patients with alosetron, a 5-HT3 receptor antagonist. AB - BACKGROUND: No currently available treatment provides consistent relief of irritable bowel syndrome. Colonic sensory and motor function are modulated partly through 5HT3-receptors. AIM: To evaluate effects of the 5HT3-receptor antagonist, alosetron, in irritable bowel syndrome. METHODS: Randomized, double-blind, placebo-controlled, dose-ranging (1, 2, 4, 8 mg b.d. alosetron), 12-week trial in 370 patients with diarrhoea-predominant or alternating constipation and diarrhoea irritable bowel syndrome. Weekly measurement of adequate relief was the key end point; other irritable bowel syndrome symptoms were collected daily using an electronic phone system. RESULTS: Alosetron (1 mg or 2 mg b.d.) significantly (P < 0.05 vs. placebo) increased the proportion of females, but not males, reporting adequate relief. Stool consistency, frequency and percentage days with urgency improved over placebo (P < 0.05) within the first month with all doses of alosetron, and persisted throughout the trial with all doses in female patients. With 1 mg b.d. alosetron, females had improved stool consistency and urgency within the first week, and adequate relief and improved stool frequency within the first 2 weeks. There was no consistent improvement in bowel function among male patients. CONCLUSION: In female irritable bowel syndrome patients with predominant diarrhoea or alternating constipation and diarrhoea, alosetron is effective in treatment of abdominal pain and discomfort and bowel-related symptoms. PMID- 10468697 TI - The effect of omeprazole dosing on the isolation of Helicobacter pylori from gastric aspirates. AB - BACKGROUND: Animal experiments suggest that omeprazole dosing increases shedding of Helicobacter into the gastric lumen, and hence into gastric juice. AIM: To assess the effect of omeprazole dosing on the yield of H. pylori from gastric aspirates of infected volunteers. METHODS: Six serial nasogastric aspirates, three before and three during dosing with omeprazole 40 mg b.d., were obtained for culture from 10 H. pylori infected volunteers and one uninfected volunteer. To reduce contamination, samples were diluted 1:10 with Maximum Recovery Diluent (MRD; pH 7.0) or HCl-KCl buffer (pH 2.2) prior to culture on Columbia and Dent's agar. RESULTS: Undiluted gastric juice cultures were rapidly overgrown by upper respiratory tract flora. HCl-KCl dilution resulted in isolation of H. pylori from 77% of infected subject aspirates before, and 67% of aspirates during dosing with omeprazole. The yields were significantly lower with MRD dilution, 47% and 10%, respectively. Omeprazole dosing significantly decreased the yield after MRD dilution, but not after HCl-KCl dilution. CONCLUSIONS: Decreasing intragastric acidity, by dosing with omeprazole, decreases the isolation of H. pylori from routinely processed gastric aspirates. In vitro acidification of gastric aspirates, by HCl-KCl dilution, increases the isolation of H. pylori both before and during omeprazole dosing. PMID- 10468698 TI - Laser assisted ratio analyser 13C-urea breath testing, for the detection of H. pylori: A prospective diagnostic European multicentre study. AB - BACKGROUND: Novel technology based on laser optogalvanic spectroscopy called the LARA (Laser Assisted Ratio Analyser) system was developed to measure 12C/13C ratios in breath samples using stable 13C isotopes, to detect Helicobacter pylori infection. AIM: To determine the sensitivity and specificity of the 13C-LARA-urea breath test in the detection of H. pylori infection in a prospective European multicentre trial; FDA-and EMEA-approved. METHODS: Consecutive dyspeptic patients underwent diagnostic gastroscopy with biopsies for culture and histopathology, to detect H. pylori infection (gold standard). Subsequently, the LARA-urea breath test was performed using either a system without a cold trap (part I) or a system with a cold trap (part II). In both instances baseline, 30-min and 60-min breath samples were collected. The optimum cut-off level for 12C/13C ratios was determined by Receiver Operator Characteristics analysis. RESULTS: In part I, 544 out of 604 patients were evaluable (low CO2: 47; withdrawn: 13). 284 out of 544 patients (52%) were H. pylori-positive according to the gold standard. The sensitivity of the LARA-urea breath test was 95% and the specificity 94%. In part II, 257 out of 272 were evaluable (low CO2: 14; withdrawn: 1). Sensitivity and specificity were 93% and 96%, respectively. CONCLUSION: The LARA-technology represents an accurate and non-invasive testing system for the detection of H. pylori infection. Its major advantages are the use of stable 13C isotope, the high throughput of samples and the easy means of collecting, storing and transporting the samples, thus making the system convenient to both patient and clinician. PMID- 10468699 TI - Sustained response to interferon-alpha or to interferon-alpha plus ribavirin in hepatitis C virus-associated symptomatic mixed cryoglobulinaemia. AB - BACKGROUND: Hepatitis C virus (HCV) infection has been associated with mixed cryoglobulinaemia. AIM: To investigate the efficacy of anti-viral therapy on the eradication of HCV and its clinical manifestations in patients with HCV associated symptomatic mixed cryoglobulinaemia. PATIENTS AND METHODS: 18 out of 32 patients with symptomatic mixed cryoglobulinaemia (MC group) received a 12 month course of interferon (3 MU three times a week, subcutaneously). Nonresponders or relapsers to this therapy were treated with interferon plus ribavirin (1200 mg/day, orally) for 12-months. 226 patients with HCV infection and without cryoglobulins were studied in comparison (Hepatitis C group). Serial quantification of serum HCV-RNA and cryoglobulins were performed. RESULTS: In the MC group, 10 out of 18 patients (55%) receiving interferon showed an end of treatment response, but at the end of follow-up, only five (28%) patients had a sustained response. In the hepatitis C group, 91 patients (47%) showed an end of treatment response but only 42 (20%) a sustained response. In the MC group alanine transaminase, cryocrit and rheumatoid factor decreased significantly in responders, with an improvement or disappearance of the MC-associated clinical manifestations. Alanine transaminase, cryocrit and rheumatoid factor increased in the relapsers and the clinical manifestations reappeared. Nonresponders and relapsers to interferon in the MC group were retreated with interferon plus ribavirin. Five out of eight nonresponders showed a end of treatment response but it was sustained in three of them. In the relapsers, treatment with combined therapy achieved a sustained response in four out of the five patients (80%). CONCLUSIONS: Interferon as monotherapy or combined with ribavirin is a safe and effective treatment in patients with HCV-associated MC. The presence of cryoglobulins does not affect the response to anti-viral treatment in patients with HCV infection. The eradication of HCV is associated with an improvement or disappearance of MC-associated clinical manifestations. PMID- 10468700 TI - Effect of ursodeoxycholic acid administration in patients with acute viral hepatitis: a pilot study. AB - BACKGROUND: Ursodeoxycholic acid (UDCA) is able to improve biochemical markers of cholestasis, with a parallel decrease in transaminases, in various cholestatic liver diseases. AIM: To evaluate the effects of UDCA administration on acute viral hepatitis-related cholestasis and the course of acute viral hepatitis. METHODS: Seventy-nine consecutive patients with acute viral hepatitis (HBV: 43, HCV: 11, HAV: 15, HEV: 3, Non A-E: 7) were randomized to receive either UDCA for 3 weeks or no treatment. Liver biochemistry and serum bile acid determinations were run at weekly intervals. RESULTS: No significant differences were observed in mean percentage decreases in transaminases between treated and untreated patients. By contrast, cholestatic indexes decreased significantly more quickly in patients treated with UDCA than in controls, and this effect was more evident in patients with increasing alanine transaminase levels at admission. After a peak at the end of the first week of therapy, serum levels of conjugated ursodeoxycholic acid (CUDCA) showed a gradual decrease. Conjugated cholic acid (CCA) and chenodeoxycholic acid (CCDCA) showed a progressive decrease with the resolution of viral hepatitis, but no influence of UDCA administration was observed. CONCLUSIONS: Our study demonstrates that UDCA significantly improves cholestatic indices in patients with acute viral hepatitis, but this effect does not seem to affect the course of the illness. PMID- 10468701 TI - The effects of oral rabeprazole on endocrine and gastric secretory function in healthy volunteers. AB - AIM: To evaluate the short-term effects of rabeprazole 20 mg on endocrine parameters, in particular serum testosterone and cortisol, and on 24 h intragastric pH, H+ activity and nocturnal gastric acid secretion. METHODS: In this double-blind, two-period crossover study, 12 healthy young male volunteers were randomly given oral rabeprazole 20 mg o.m. or placebo for 14 days. There was a washout period of at least 1 week between the two studies. The effects of rabeprazole and placebo on cortisol and testosterone (primary criteria), and on tri-iodothyronine, thyroxine, 17beta-oestradiol, thyroid-stimulating hormone, thyroxine-binding protein, parathyroid hormone, insulin, glucagon, rennin, aldosterone, follicle-stimulating hormone, luteotrophic hormone, prolactin, somatotrophic hormone, dehydroepiandrosterone, cortisol-binding globulin and urinary 6-beta hydroxycortisol were compared. Intragastric 24 h pH, 24 h H+ activity and nocturnal gastric acid secretion were determined by pH probe and gastric aspiration. RESULTS: Rabeprazole produced no clinically relevant effects on endocrine function as assessed by measurement of serum testosterone, circadian serum cortisol levels, ACTH-stimulated serum cortisol levels and 17 other endocrine function tests. Rabeprazole significantly increased the 24 h median pH values compared to placebo (on Days 7 and 14 median values ranged from 3.92 to 6.88 with rabeprazole and from 1.48 and 4.22 with placebo, P < 0.001) and significantly decreased the integrated 24 h H+ activity (AUC08--08) from 343 mmol/L/h with placebo to 44 mmol/L/h with rabeprazole (P < 0.001). Following cessation of dosing, intragastric pH levels decreased and H+ activity increased, but acid secretion did not recover completely during the next 72 h. The mean value for nocturnal gastric acid secretion on Days 7 and 8 was 36 mmol/6 h with placebo and 5.6 mmol/6 h with rabeprazole (P < 0.001). Rabeprazole was well tolerated. CONCLUSION: Rabeprazole did not influence endocrine function in healthy young male volunteers during short-term dosing. Rabeprazole substantially increased intragastric pH over a 24 h period and significantly decreased intragastric acidity and nocturnal gastric acid secretion. PMID- 10468702 TI - Regression of columnar-lined (Barrett's) oesophagus with omeprazole 40 mg daily: results of 5 years of continuous therapy. AB - BACKGROUND: We have previously reported the effect of 2 years of omeprazole 40 mg daily on columnar-lined (Barrett's) oesophagus (CLO). AIMS: In the present study, follow-up has been extended to 5 years to assess the macroscopic and microscopic effects of continuing therapy. PATIENTS AND METHODS: The 23 patients have been followed for up to a further 3 years. Endoscopy with multiple biopsies was performed at the end of years 3, 4 and 5. RESULTS: Although there had been a statistically significant regression in the length of CLO after 2 years, there was no overall further measurable change after 5 years. However, one patient showed complete macroscopic and microscopic regression. The number and size of macroscopic squamous islands within the CLO continued to increase, and there was a further increase in microscopic squamous re-epithelialization of surface mucosa, gland ducts and Barrett's gland tissue. Low-grade dysplasia was found consistently in one patient in biopsies taken up to the end of year 3 but it could not be detected thereafter. CONCLUSIONS: Omeprazole 40 mg daily appears to have beneficial effects on CLO, although it rarely induces a complete regression. Whether the benefits will reduce the risk of malignant transformation is unknown. PMID- 10468703 TI - Post-prandial ranitidine is superior to post-prandial omeprazole in control of gastric acidity in healthy volunteers. AB - BACKGROUND: Episodic heartburn is a common problem, affecting over 40 million Americans. Although omeprazole provides excellent acid suppression when used daily, the use of omeprazole as on-demand therapy for episodic symptoms has not been extensively studied. AIM: To compare the onset and duration of post-prandial gastric acid suppression by omeprazole (10 and 20 mg) or ranitidine (75 and 150 mg) taken as single doses in healthy volunteers. METHODS: Twenty-four healthy volunteers (14 male, 10 female, mean age 33.4 years, range 18-56 years) were given ranitidine (RAN) 75 mg or 150 mg vs. omeprazole (OME) 10 mg or 20 mg when pH returned to below 2 after breakfast, in a randomized open label crossover design, with a washout of at least 2 days between medications. Intragastric pH was monitored for 6 h. The time between drug ingestion and rise of gastric pH > 3 and 4, and total time pH remained > 3 and 4 during the 6 h post drug, was compared between groups using two way ANOVA and Wilcoxon matched pairs test. RESULTS: The median time needed to pH > 4 was 204.5 min for RAN 75 mg, 186 min for RAN 150 mg and > 360 min for both OME 10 and 20 mg (P < 0.001 between the four groups). The median time that pH remained > 4 was 93 min for RAN 75 mg, 143.5 min for RAN 150 mg and 0 min for both OME 10 and 20 mg (P < 0.001 between the four groups). Both doses of RAN were significantly superior to both doses of OME, although no significant difference was found between the high and the low doses of either drugs. Similar results were found for pH > 3. CONCLUSION: Ranitidine 75 or 150 mg provides more rapid increase in gastric pH to > 3 and > 4 compared to omeprazole 10 or 20 mg when taken at the end of the post-prandial period. These data suggest that ranitidine may be more effective for episodic post-prandial heartburn than omeprazole. PMID- 10468704 TI - Lansoprazole pharmacokinetics differ in patients with oesophagitis compared to healthy volunteers. AB - AIM: To compare the pharmacokinetics of lansoprazole in patients with reflux oesophagitis and in healthy volunteers, after a single dose and at steady-state. PATIENTS AND METHODS: A 30 mg dose of lansoprazole was administered orally daily for 7 days in eight healthy male volunteers aged 21-24 years, and in 16 patients aged 29-65 years with grade 2 or 3 reflux oesophagitis. The pharmacokinetics were assessed over the 24 h dose interval following the first dose and again after the 7th dose. RESULTS: Within both the patient and volunteers groups, there were no significant differences between day 1 and day 7 in any of the pharmacokinetic parameters including maximum concentration (Cmax), area under the concentration time curve (AUC), and terminal half-life of elimination (t(1/2)). However, on both days 1 and 7, values were significantly higher in the patients than in the healthy volunteers. On day 7, Cmax was 1343 ng/mL in patients compared with 765 ng/mL in healthy volunteers, AUC was 3458 ng.h/mL vs. 1350 ng.h/mL and t(1/2) was 1.62 h vs. 0.90 h. CONCLUSION: The differences in results for the pharmacokinetics reflect reduced lansoprazole clearance in the patient group. Other research has not found a difference in pharmacokinetics when comparing healthy volunteers with patients with acid-related disorders. The difference in lansoprazole clearance in this study may be related to a variety of factors that are different in patients compared with young normal volunteers, such as age, gender, other drugs, and reduced general well-being. PMID- 10468705 TI - Early experience with intrasphincteric botulinum toxin in the treatment of achalasia. AB - BACKGROUND: Recent reports have suggested that intrasphincteric injection of botulinum toxin is effective and long-lasting in the treatment of achalasia. AIM: To report our experience of botulinum toxin injection in a prospective series of consecutive patients with achalasia. METHODS: Eleven consecutive patients with achalasia (eight male, mean age 55 years, range 20-87) were treated with 60 units of botulinum toxin (Dysport; Speywood Pharmaceuticals Ltd, UK) into each of four quadrants at the lower oesophageal sphincter. Patients were assessed pre treatment and 1 month after treatment using a symptom score and oesophageal manometry. Median follow-up was 12 months (range 6-28). RESULTS: The injection procedure was simple to perform and free of adverse effects. Although treatment had a beneficial effect on dysphagia (median pre-treatment score 3 [interquartile range 3-3]; post-treatment score 2 [0-3]: P=0.03) 1 month following therapy, there was no significant improvement in chest pain or regurgitation scores. Similarly, no significant reduction in median lower oesophageal sphincter pressure was observed (29.5 mmHg [21-42] pre-treatment, 28.5 [17.5-55.5] post treatment P=0.67). Four patients (36%) required further therapy within 3 months and the overall relapse rate was 73% (eight of 11) within 2 years. CONCLUSION: Although botulinum toxin injection was well tolerated, these results using Dysport at a dose of 240 mouse units question its efficacy as a treatment for achalasia. PMID- 10468706 TI - Ibuprofen augments gastrointestinal symptoms in lactose maldigesters during a lactose tolerance test. AB - BACKGROUND: Clinical symptoms during lactose tolerance test mimic those seen after therapeutic administration of prostaglandins, and resemble inflammatory processes. AIM: To investigate the possibility that lactose-induced gastrointestinal symptoms are associated with prostaglandins and/or nitric oxide. METHODS: After an overnight fast, nine maldigesters ingested lactose or sucrose with or without an inhibitor of prostaglandin synthesis (ibuprofen), in a randomised double-blind crossover trial. Gastrointestinal symptoms, concentrations of PGE2-M in blood and urine, and urinary 6-keto PGF1alpha (as indicators of prostaglandin synthesis), and urinary nitrate and nitrite as well as cyclic GMP excretions (as indicators of nitric oxide formation), were measured. RESULTS: Ibuprofen increased the first 3-h symptom scores (flatulence + borborygmi + abdominal bloating + pain) caused by lactose (P=0.008) but not sucrose. The concentrations of PGE2-M in the plasma and in the urine were unaffected. Lactose increased the urinary excretion of 6-keto PGF1alpha by about 30% (P=0.17), which was inhibited by ibuprofen (P=0.02). The production of nitric oxide was unaffected by lactose or ibuprofen. CONCLUSION: The inhibition of prostaglandin synthesis intensified gastrointestinal symptoms in lactose maldigesters, suggesting a negligible role for prostanoids in lactose-induced symptoms. PMID- 10468707 TI - Reduction of the nociceptive response to gastric distension by nitrate ingestion in rats. AB - BACKGROUND: Dietary nitrates are known to produce nitric oxide in the stomach, which may influence gastric function. AIM: To investigate whether nitrate ingestion modifies gastric sensitivity to distension through a mechanism involving nitric oxide production. METHODS: Nociception, associated with gastric distension ranging from 10 to 40 mmHg, was assessed in anaesthetized rats by the amplitude of cardiovascular depressor responses. Gastric volume corresponding to each distension was recorded. The following intragastric administrations (1 mL) were performed before distension: water (control), KNO3, NaNO3, KCl, NaCl (all at 0.1 mmol/kg), standard food (0.5 g), sodium nitroprusside, a nitric oxide donor (5 mg/kg), and haemoglobin, a nitric oxide scavenger (150 mg/kg) given either with water or KNO3. RESULTS: In controls, the fall in blood pressure increased from 7.8 +/- 2.0 to 31.6 +/- 2. 7 mmHg at distending pressures from 10 to 40 mmHg, respectively. KNO3 significantly reduced the amplitude of blood pressure response for the highest distending pressures (35 and 40 mmHg), while KCl induced a reduction in blood pressure response at all gastric pressures. NaNO3 and NaCl did not induce significant changes in distension-induced depressor responses. Administration of 0.5 g of standard food or sodium nitroprusside reproduced the effect of KNO3, which was reversed by haemoglobin. None of the compounds modified the gastric pressure-volume relationship, except KNO3, which increased gastric volume for the lowest distending pressures, and haemoglobin, which reduced the volume for the highest pressure. CONCLUSIONS: Ingestion of potassium nitrate reduces the sensitivity to gastric distension, through a mechanism involving nitric oxide. PMID- 10468708 TI - Dilatation and constriction of rat gastric mucosal microvessels through prostaglandin EP2 and EP3 receptors. AB - BACKGROUND: Prostaglandin (PG)E2 has both a vasodilating action and a protective function in the gastric mucosa. There are four subtypes of PGE2-sensitive, or EP, receptors. AIM: To identify the subtype of EP receptors in the microvessels of the rat gastric mucosa using EP2 and EP3 receptor agonists. METHODS: The posterior wall of the anaesthetized rat stomach was secured in a chamber and superfused with Tyrode's solution, and the gastric microcirculation of the mucosal base was observed through a window with transillumination. PGE2 and its derivatives (20 microL) were applied topically in the window. RESULTS: PGE2 (0.001-10 micromol/L), misoprostol (EP2/EP3 receptor agonist; 0.01-100 micromol/L) and butaprost (EP2 receptor agonist; 1-1000 micromol/L) dilated the arterioles dose-dependently, but M&B 28 767 (EP3 receptor agonist; 0.001-10 micromol/L) did not alter their diameters. M&B 28 767 constricted the venules and collecting venules dose-dependently whereas butaprost dilated them. PGE2 and misoprostol had bell-shaped dose-response curves: constriction by low doses of PGE2 and misoprostol (0.001-0.1 micromol/L and 0.01-1 micromol/L) and dilation by high doses of PGE2 and misoprostol (0.1-100 micromol/L and 1-100 micromol/L). CONCLUSIONS: These results suggest that PGE2 dilated both arterioles and venules in the rat gastric mucosa through the EP2 receptors and constricted the venules through the EP3 receptors. PMID- 10468709 TI - Efficacy of 1-week ranitidine-bismuth-citrate (RBC)-based triple therapy for eradication of Helicobacter pylori infection. PMID- 10468710 TI - 5-day H. pylori eradication. PMID- 10468713 TI - Urothelial function. PMID- 10468712 TI - The role of obesity and diet in urological carcinogenesis. PMID- 10468714 TI - Lithogenic risk factors in normal black volunteers, and black and white recurrent stone formers. AB - OBJECTIVE: To compare lithogenic risk factors in normal black volunteer men (BN), male black stone formers (BSF) and male white recurrent stone formers (WSF); in addition, the differential diagnoses in the stone formers were compared to determine if the causes of renal stones differed in the two groups. SUBJECTS AND METHODS: The study included 22 BN, 22 consecutive BSF and 122 consecutive WSF seen over a 10-year period. Each subject was assessed by a thorough medical history, dietary analysis and a full serum and urinary biochemical evaluation. RESULTS: Although the WSF were significantly more overweight (P<0.001) and obese (P<0.001) than the BSF, neither group had values significantly different from those of their respective normal populations. There was no significant difference in serum levels of Ca, phosphorus, ionised Ca, calcitriol and alkaline phosphatase in BSF and WSF, but levels of parathyroid hormone tended to be (P<0.1) and calcitriol was significantly higher in BSF than WSF (P<0.03). The BSF urinary excretion levels tended to be more like that of the WSF, with the following pattern present; BN/=0.70 and >/=0.06, respectively. The sensitivity, specificity and overall accuracy of RI and DeltaRI values before and after giving diclofenac were calculated using IVU as the reference standard. RESULTS: IVU showed that both kidneys were normal in 20 patients that 40 had unilateral obstruction. At baseline, the mean (sd) RI was 0.72 (0.06) in the 40 obstructed kidneys, significantly higher than the RI of 0.60 (0.06) for the 60 normal kidneys (P<0. 001). Six hours after giving diclofenac, the mean RI of normal kidneys remained stable and that of the obstructed kidneys decreased significantly to 0.67 (0.06) (P<0.001) and was almost stable at 12 h. Nevertheless, the mean RI of the obstructed kidneys remained significantly higher than that of the normal kidneys at 6 and 12 h after diclofenac. The mean DeltaRI decreased significantly from 0.11 (0.04) at baseline to 0.06 (0.04) at 6 h after diclofenac (P<0.001) and remained almost stable at 12 h. In the 10 patients undergoing DU after the relief of obstruction, the mean RI returned to normal. At baseline, RI was sensitive in 70% and specific in 98%, while DeltaRI was sensitive in 88% and specific in all cases. Six hours after diclofenac, the specificity of RI and DeltaRI remained stable while the sensitivity decreased significantly to 50% and 68%, respectively. CONCLUSION: NSAIDs significantly decrease the RI of acutely obstructed kidneys, causing a significant reduction in the sensitivity of DU values identifying acute renal obstruction. To evaluate acute loin pain, DU should be carried out while the patient is in pain and before giving NSAIDs. PMID- 10468716 TI - Endourological renal salvage in patients with calculus nephropathy and advanced uraemia. AB - OBJECTIVE: To document our experience with percutaneous nephrolithotripsy (PCNL) for the treatment of calculus nephropathy and advanced renal failure. PATIENTS AND METHODS: Between October 1989 and April 1996, 78 patients with calculus nephropathy (51 males and 27 females, mean age 42 years, range 16-67; mean serum creatinine level 663.8 micromol/L, range 282.8-1449.7) were treated in our department. Forty-six of the patients had bilateral upper urinary calculi, while the remaining 32 had a solitary functioning kidney obstructed by stone. Thirty two patients had a normal urine output, 26 were oliguric and 20 presented with anuria. A preliminary percutaneous nephrostomy (PCN) was performed under local anaesthesia in 64 patients, and stone retrieved subsequently through the same nephrostomy track after improvement in their uraemic status. In the remaining 14 patients, the stones were removed at the first session. Of the 46 patients with bilateral stones, 21 underwent stone retrieval by PCNL on the contralateral side after an interval of 2-3 weeks, while 25 patients underwent simultaneous JJ stenting on the contralateral side, followed, after improvement in uraemic status, by delayed PCNL or extracorporeal shock wave lithotripsy on that side. RESULTS: There were three deaths, caused by overwhelming sepsis; 64 of the remaining 75 patients recovered well from uraemia, while 11 showed no improvement, suggesting irreversible renal damage. Complications included five patients with nonfatal sepsis and five with secondary haemorrhage. At the last follow-up 2.5-9 years after presentation, the mean serum creatinine level (271.3 micromol/L) was significantly better than the mean preoperative level. Six patients are on maintenance haemodialysis. CONCLUSIONS: It is not uncommon in developing countries for patients with urolithiasis to present with advanced uraemia caused by long-standing obstruction, almost invariably complicated by infection. A percutaneous endourological approach offers the best hope for these patients, and in our experience a significant percentage of these cases can be salvaged. PMID- 10468717 TI - Flexible ureterorenoscopy for the treatment of refractory upper urinary tract stones. AB - OBJECTIVE: To report our experience of using flexible ureterorenoscopy for upper urinary tract stones. PATIENTS AND METHODS: Thirty-seven patients (13 females, 24 males, mean age 48 years, range 10-76) with upper urinary tract stones not responding to other treatments had their residual symptomatic stones treated through a 9.3 F flexible ureterorenoscope. Electrohydraulic lithotripsy (EHL) was used to fragment the stones and a basket or a grasper used to retrieve the fragments. RESULTS: Twenty-three patients were completely freed of stone and a further five had asymptomatic residual fragments of < 5 mm. Of 15 patients with ureteric stones, 14 were rendered stone-free at the end of the procedure and 14 of the 22 patients with kidney stones were treated successfully; 80% of the lower calyceal stones were treated successfully and a firm decision was possible on the further management of the remaining stones. There were no major complications. CONCLUSION: Flexible ureterorenoscopy should be considered in patients with symptomatic upper ureteric or renal stones refractory to other treatments, but the instrumentation is expensive and delicate, and its use should be reserved for such cases. PMID- 10468718 TI - Electrokinetic lithotripsy: safety, efficacy and limitations of a new form of ballistic lithotripsy. AB - OBJECTIVE: To investigate the safety and efficacy of electrokinetic lithotripsy (EKL), a ballistic lithotripter which uses high-energy magnetic fields to propel an impactor to fragment calculi. PATIENTS AND METHODS: The records and radiographs of 121 patients who underwent ureteroscopy using the EKL for stones in the upper (26), mid (28) or lower (67) ureter were reviewed retrospectively. Ureteroscopy was performed with an 8.5 F semi-rigid ureteroscope, through which a 3 F EKL probe was passed. RESULTS: A total of 148 stones (mean stone size 11.5 mm, range 6-40) in 121 patients were treated using the EKL. One patient was lost to follow-up. Of 148 stones, 147 (99.3%) were fragmented, including five that had resisted fragmentation with either pulsed-dye laser or electrohydraulic lithotripsy. Despite this, only 45 of 56 patients (80%) with a single stone in the lower ureter were rendered stone-free after a single ureteroscopic procedure. Seven patients in this group (12%) required shock-wave lithotripsy for fragments that had been propelled into the kidney, while four patients (7%) required repeat ureteroscopy for retained ureteric fragments. Complications were limited to minor ureteric perforations in two patients, both of which were treated with a stent. CONCLUSION: EKL is an inexpensive and reliable endoscopic method which fragments nearly all urinary calculi. Its limitations include the propulsion of fragments and the need to use an offset, semi-rigid ureteroscope. We recommend the use of a basket or graspers to remove fragments of >/=4 mm after EKL. PMID- 10468719 TI - A comparative analysis of nephrostomy, JJ stent and urgent in situ extracorporeal shock wave lithotripsy for obstructing ureteric stones. AB - OBJECTIVES: To determine the optimal method of treatment for ureteric stones causing complete obstruction, treated by insertion of a JJ stent or a nephrostomy tube, followed by extracorporeal shock wave lithotripsy (ESWL) or by urgent in situ ESWL if readily available. PATIENTS AND METHODS: The study comprised a retrospective analysis of 82 consecutive patients who presented with ureteric stones causing complete obstruction. Twenty-six had a percutaneous nephrostomy (PCN, group 1) and 40 had a JJ stent (group 2) placed to relieve the obstruction, and the stones were subsequently treated by ESWL. Sixteen patients underwent urgent in situ ESWL without recourse to either a JJ stent or a PCN (group 3). The choice of the procedure was not determined by stone size, site or other factors, but mainly by the attending surgeon's preference or the availability of urgent ESWL. The success rate was measured by the disintegration of the stone and spontaneous passage after ESWL; failure was defined as the need for additional procedure(s) for stone extraction. RESULTS: Urgent in situ ESWL (group 3) had a median (95% confidence interval) success rate of 81 (54-96)%, compared with 70 (53-83)% in group 2 and 54 (33-73)% in group 1. CONCLUSION: If facilities are available, urgent in situ ESWL appears to be the choice of treatment for obstructing ureteric stones. If such facilities are not available, a JJ stent may offer better success than a PCN. A prospective controlled trial is necessary to confirm these findings. PMID- 10468720 TI - The conservative management of corynebacterium group D2 encrusted pyelitis. AB - OBJECTIVE: To evaluate the conservative management of encrusted pyelitis (EP), an infectious disease caused by urea-splitting micro-organisms (Corynebacterium group D2 being the most common), and characterized by stone encrustation of the collecting system. PATIENTS AND METHODS: Four patients (aged 17-62 years) with EP caused by Corynebacterium group D2 were treated conservatively with intravenous vancomycin and percutaneous acidification of the renal collecting system. Percutaneous nephrostomy tubes were placed to allow irrigation with Thomas' acid solution, the outflow being ensured by ureteric catheters. Patients were monitored and the results of and tolerance to treatment evaluated clinically, by biological assays, and by imaging techniques (ultrasonography, computed tomography and pyelography). RESULTS: No kidneys were lost and in all patients with initial renal failure, creatinine levels recovered to previous values. The adverse effects were moderate, consisting of flank pain or discomfort, and low grade fever. Three patients developed asymptomatic fungal urinary tract infections requiring antifungal agents. The encrustations were treated effectively and almost disappeared in two patients, while the other two had no residual calcification. One transplanted-kidney recipient developed a ureteric stricture requiring surgical repair. Within a mean follow-up of 16 months, there was no recurrence of EP and the residual encrustation remained small. CONCLUSION: The conservative treatment of EP can avoid surgery in many cases, although there are some adverse effects of irrigation. The long-term follow-up and prevention of recurrent EP are nevertheless required. PMID- 10468721 TI - Symptom characteristics and the development of tolerance with time in patients with indwelling double-pigtail ureteric stents. AB - OBJECTIVE: To determine the symptoms of and factors predicting the tolerance to double-pigtail ureteric stents, and the development of tolerance with time. PATIENTS AND METHODS: The study included 39 patients (median age 49.0 years, range 26-74; 24 men and 15 women) who were treated in our department for ureteric obstruction caused by benign conditions, excluding pregnancy and associated open surgery. All patients received a polyether-urethane double-pigtail ureteric stent (7 F, 28 cm long). Tolerance to the procedure was assessed using a questionnaire and a 10-cm linear visual analogue scale (VAS) at 24 h and again one week after placement, and on the day before stent removal. RESULTS: From the end of the first week to before removal of the stent, fewer patients reported dysuria, haematuria and having recourse to painkillers than during the first week. However, analysis of variance for repeated measures showed no significant difference between the VAS scores for first day, the first week and the day before removal, either overall or stratified by gender (P=0.15). A factorial analysis of variance analysing the VAS score for the first week as a function of gender, age and type of occupation (sedentary or mobile occupation) showed a significant effect only for gender (P=0.005) and gender-age interaction (P=0. 02): VAS scores were higher in men and particularly in younger men. CONCLUSION: Almost all patients with short-term placement of indwelling double-pigtail ureteric stents have untoward symptoms. Although some symptoms, e.g. dysuria and haematuria, significantly improve with time, the general tolerance remains unchanged. Tolerance in men, and particular in younger men, was significantly poorer. PMID- 10468722 TI - The impact of contemporary bladder management techniques on struvite calculi associated with spinal cord injury. AB - OBJECTIVE: To evaluate the incidence, risk factors and complications of upper tract struvite calculi, often associated with spinal cord injury (SCI), as such patients have a high incidence of urinary infection complicating their neurogenic voiding dysfunction, by reviewing a large population of patients with SCI in whom modern techniques of bladder management were used. PATIENTS AND METHODS: Between 1982 and 1996, 1669 patients with SCI were admitted to our institution; 1359 of these patients sustained their injuries during the study period. During this time, their bladder management was based on urodynamic and imaging criteria, using techniques such as early intermittent catheterization, sphincterotomy and bladder augmentation where possible to create a catheter-free, low-pressure reservoir. All instances of upper tract struvite calculi in this population were documented. Risk factors for stone development, presentation and complications, management and recurrence rates were assessed. RESULTS: Over the 15 years, 58 patients (3.5% of the SCI population) were treated for a total of 144 episodes of struvite calculi. The incidence of stones in those injured since 1982 was 1. 5%; 67% of these patients had complete spinal cord lesions, 54% had lesions of the cervical cord and 53% developed their first stone >10 years after injury. Only 22% presented within 2 years of injury. The group of patients developing stones had a significantly higher incidence of indwelling catheters (49%), bladder stones (52%) and vesico-ureteric reflux (28%) than those who were stone-free. The development of recurrent urinary tract infections was the most common mode of presentation. The stone-free rate after treatment was 87%. Normal renal function was preserved in 72% of patients. CONCLUSIONS: In a large population of patients with SCI managed using contemporary bladder techniques the incidence of upper tract calculi was 3.5%; 30% of these stones were complete or partial staghorns. Those patients with complete cord lesions, permanent indwelling catheters and vesico-ureteric reflux were at the highest risk. Stone clearance was 87% and recurrent stones occurred in 69% of patients. Struvite renal calculi continue to be a significant problem in the spinal cord injury population. PMID- 10468723 TI - The epidemiology of benign prostatic hyperplasia: a study in Greece. AB - OBJECTIVE: To assess the epidemiology of benign prostatic hyperplasia (BPH) in a case-control study in greater Athens, Greece. PATIENTS AND METHODS: The study comprised 184 patients surgically treated for BPH within one year of its diagnosis (cases) and 246 patients with no symptoms of BPH who were treated in the same hospitals for minor diseases or conditions (controls). All cases and controls were permanent residents of the greater Athens area, Greece. The data were assessed using unconditional logistic regression. RESULTS: After controlling for age and education, cases and controls had similar distributions for height, body mass index, sibship size and birth order in the parental family, marital status, number of offspring and a series of previous medical diagnoses or surgical operations. The sole exception was surgery for haemorrhoids, that appeared to be related to the incidence of BPH, possibly by chance. There was no evidence that vertex baldness, tobacco smoking, alcohol consumption or coffee drinking increased the risk for BPH. Men who had spent most of their lives in a rural rather than an urban environment appeared to be at reduced risk for BPH. CONCLUSION: The lifestyle factors assessed here have no major effect on the aetiology of BPH. PMID- 10468724 TI - Sham treatment compared with 30 or 60 min of thermotherapy for benign prostatic hyperplasia: a randomized study. AB - OBJECTIVE: To study the differences in subjective and objective results after 30 or 60 min of transurethral thermotherapy (TUMT) or sham treatment in men with benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: Forty-four patients with lower urinary tract symptoms caused by BPH were randomized to undergo 30 or 60 min of TUMT or sham treatment (14, 16 and 14 patients, respectively). The patients were evaluated using symptom scores, timed micturition, free flow rates and urodynamics before and after treatment. They were followed for 1 year, at the end of which they rated the treatment results on a self-administered visual analogue scale. RESULTS: Forty-two patients completed the study. The treatment failed in some patients in all groups, but the improvement in maximum and median flow rates, timed micturition and micturition frequency (day and night) was greater after TUMT than in the sham-treated group. Symptom scores also improved more after TUMT, although not significantly. There was a good correlation between the decrease in urinary frequency, especially nocturnal, and the patients' perception of a successful treatment, whereas the increase in maximum urinary flow rate was apparently of little importance. CONCLUSIONS: TUMT had a better effect than the placebo treatment, having a greater impact on irritative than on obstructive symptoms. More patients were satisfied after the 30-min than after the 60-min treatment. PMID- 10468725 TI - Mortality in the elderly in relation to nocturnal micturition. AB - OBJECTIVE: To study the relationship between nocturnal micturition and mortality in an elderly population. SUBJECTS AND METHODS: All 10 216 members of the Swedish pensioners' association (SPF) in two Swedish counties were asked to participate in a questionnaire survey. The questions concerned the general state of health, occurrence of somatic diseases and symptoms, number of voiding episodes per night, everyday habits and behaviour, and the use of drugs. After 6 years, data on deaths were extracted from the National Register of Deaths at the National Central Bureau of Statistics in Stockholm. RESULTS: There were 6143 evaluable questionnaires, of which 39.5% were from men. The mean (sd) ages of the men and women participating were 73.0 (6.0) and 72.6 (6.7) years, respectively. During a 54-month period after the questionnaire was completed, 444 of the men and 384 of the women died. The men with three or more nocturnal voiding episodes had a higher death rate, at 1.9 (1.4-2.6) times more than the whole group of men (3.4% vs 1.9% per 6 months; P<0.001), and the corresponding women a death rate 1.3 (0.9 2.0) times higher than all women (1.4% vs 1.1% per 6 months, not significant). In a multiple logistic regression analysis, significant independent correlates of death during the study period were: being 70-79 years vs <70 years (odds ratio, OR, 1.92, 95% CI 1.52-2.43), being >/=80 years vs <70 years (5.14, 3.96-6.68), female gender (0.44, 0.37-0.52), health (1. 81, 1.46-2.25), negative health development (1.70, 1.40-2.06), spasmodic chest pain (1.38, 1.09-1.74), diabetes (1.59, 1.18-2.15), stroke (1.82, 1.12-2.93), and three or more vs two or fewer voiding episodes (1.34, 1.01-1.79). CONCLUSION: Elderly people who need to void three or more times per night had a greater mortality rate over a 54-month observation period than those voiding less often. PMID- 10468726 TI - The effect of colonoscopy on serum prostate specific antigen levels. AB - OBJECTIVE: To evaluate, in a prospective trial, the effect of colonoscopy on serum prostate specific antigen (PSA) levels (which are known to increase after cystoscopy and prostatic needle biopsy) in men aged 50-70 years, as they commonly undergo colonoscopy and PSA screening together. SUBJECTS AND METHODS: Twenty-four consecutive men (mean age 61 years, range 37-78) were enrolled in the study. All subjects had their serum PSA measured 24-72 h before undergoing colonoscopy. Further values were obtained 24 h, 7 and 30 days after the procedure. The values before and after colonoscopy were compared to evaluate the effect of colonoscopy on serum PSA levels. RESULTS: Of 24 patients, seven (35%) had insignificant increases at 24 h after colonoscopy; 12 (50%) patients had an insignificant increase in PSA level 30 days after the procedure. None of the 24 patients had a significant increase after colonoscopy when compared with baseline values. CONCLUSION: Flexible colonoscopy does not adversely affect serum PSA levels. Although individuals may have a 30% daily fluctuation in serum values, this study detected no significant change in serum PSA levels after prolonged transrectal prostatic manipulation with a colonoscope. PMID- 10468727 TI - Intraoperative electrical stimulation of cavernosal nerves with monitoring of intracorporeal pressure in patients undergoing nerve sparing radical prostatectomy. AB - OBJECTIVE: To explore the utility of intraoperative cavernosal nerve stimulation in facilitating atraumatic nerve dissection during radical prostatectomy, and thus help predict postoperative erectile function. PATIENTS AND METHODS: Fourteen patients (aged 51-72 years) underwent nerve-sparing radical retropubic prostatectomy (NSRRP); 10 were potent before surgery (group 1), and four had erectile dysfunction (group 2). A multi-acquisition system (MacLab/8e) with a Macintosh computer was used for real-time display and recording of intracavernosal pressure (ICP) during surgery. Nerves were stimulated with a bipolar probe (monophasic rectangular pulses, 10 mA, 20 Hz, 0.22 s) before and after removal of the gland. The follow-up consisted of interviews with patients and their partners' 12-18 months after treatment. RESULTS: The mean (sem) basal ICP of 8. 0 (2.0) cmH2O remained unchanged during nerve dissection. The mean increase in ICP during electrical stimulation was >50 cmH2O in seven potent patients (group 1) and was sustained as long as the nerve was stimulated. Postoperatively, these seven patients reported erections sufficient for sexual intercourse. However, the three remaining patients in group 1 had pressure rises of <30 cmH2O, of whom two reported partial erections and one reported total impotence postoperatively. The recovery time for erectile function was 6-12 months after surgery. Two patients from group 2 had transient increases in ICP to <40 cmH2O; one had an increase to 20 cmH2O and one had no response at all. All four patients remained totally impotent postoperatively. There were no complications. CONCLUSIONS: Intraoperative electrical stimulation of the cavernosal nerves with ICP monitoring before and after NSRRP is a safe and reliable method for documenting nerve continuity and its functional status. Patients who have normal preoperative erectile function and show an adequate rise in ICP upon electrical nerve stimulation during NSRRP will almost certainly be potent after surgery. This tool may be used to facilitate atraumatic nerve dissection during NSRRP. PMID- 10468728 TI - A population-based study of clinical and pathological prognostic characteristics of men with familial and sporadic prostate cancer. AB - OBJECTIVES: To compare traditional prognostic characteristics of familial vs sporadic prostate cancers and to investigate potential detection biases arising from differences in the use of screening and investigative procedures. PATIENTS AND METHODS: Familial and sporadic cancers were identified in a population-based sample of incident prostate cancers (total 318) in Auckland, New Zealand. To examine the potential for detection biases in these comparisons, the sociodemographic and clinical characteristics were determined according to family history status for a sample of 959 patients newly referred to Auckland urology clinics by general practitioners for the investigation of prostate-related conditions. RESULTS: Compared with sporadic prostate cancers, familial cancers were more likely to be diagnosed in patients at a younger age (P=0.05), after asymptomatic serum prostate-specific antigen (PSA) screening (P=0. 02), and to include a lower proportion with extraprostatic disease (P=0.009) and serum PSA levels before diagnosis of >20 ng/mL (P=0. 04). This was consistent with the observed trend for patients referred to urology clinics with a positive family history to be of higher socio-economic and educational status and to more frequently undergo screening and biopsy investigation. CONCLUSION: Familial prostate cancers appeared to be diagnosed at an earlier stage of disease progression in this study population, possibly as the result of the higher socio economic status and greater use of screening and investigative procedures amongst patients reporting a positive family history. These features reduce the validity of cross-sectional comparisons of prognostic variables for familial vs sporadic prostate cancer and emphasize the need for further longitudinal prognostic studies. PMID- 10468729 TI - Prediction of prostatic cancer progression after radical prostatectomy using artificial neural networks: a feasibility study. AB - OBJECTIVE: To report a methodological feasibility study in a small series of patients with node-negative organ-confined prostatic cancer, using artificial neural networks to predict tumour progression after radical prostatectomy and thus help to identify high-risk patients who would benefit from adjuvant treatment. PATIENTS AND METHODS: A group of 20 patients with pT2N0 prostatic cancer and postoperative tumour progression was compared with a control group of 20 patients with no progression, matched for age, duration of follow-up and preoperative serum prostate-specific antigen level. Histopathological data were obtained from the radical prostatectomy specimens, i.e. the Gleason score, World Health Organisation (WHO) grade and maximum diameter of the tumour transects. The volume and surface area of the epithelial tumour component and of the lumina of the neoplastic glands per unit tissue volume were estimated by morphometric methods. To predict recurrence, multilayer feedforward networks with backpropagation (MLFF-BP), two implementations of learning vector quantization (LVQ), and linear discriminant analysis (LDA) were applied. The ability of these models to correctly classify new cases was tested using the 'leave-one-out' technique. RESULTS: Progression was predicted correctly in 85% of newly presented cases from the three routine histopathological variables alone. On the basis of the four morphometric variables alone progression was predicted correctly in 93% of cases. The use of all seven variables as input data only slightly improved the quality of prediction. The best results were obtained by the LVQ networks and LDA, followed by MLFF-BP networks. CONCLUSIONS: In this methodological feasibility study, the progression of pT2N0 prostatic cancer after radical prostatectomy could be predicted with good accuracy, sensitivity and specificity from routine variables or morphometric texture variables using artificial neural networks. These results suggest that this approach should be assessed in a prospective study with more cases. PMID- 10468730 TI - Laparoscopy as the investigation and treatment of choice for urinary incontinence caused by small 'invisible' dysplastic kidneys with infrasphincteric ureteric ectopia. AB - OBJECTIVE: To report our experience of using laparoscopy for the accurate location and simultaneous removal of small dysplastic kidneys with ectopic ureteric insertion causing urinary incontinence and that were not detected by conventional imaging modalities. PATIENTS AND METHODS: Seven girls (mean age 7.9 years, range 3.5-13) presented with urinary leakage occurring between normal voids. Imaging studies including ultrasonography, renal scintigraphy, intravenous urography, computed tomography and/or magnetic resonance imaging in six of the seven patients revealed a single normal functioning kidney, but failed to detect the contralateral nonfunctioning dysplastic kidney. All patients were examined under anaesthesia, followed by transperitoneal laparoscopy for the simultaneous localization and removal of the dysplastic kidneys under the same setting. RESULTS: Laparoscopy in all seven patients revealed a small dysplastic kidney that could always be easily located by first finding the draining ureter over the iliac vessels and then following it upwards. Four dysplastic kidneys were found in the renal fossa (two left, two right). One kidney was found at the left iliac fossa just above the pelvic brim, one at the left lumbar region, and the other at the right iliac fossa. Laparoscopic nephroureterectomy was successful in all seven girls and the patients were discharged 48 h after surgery. The follow-up (mean 2.7 years, range 3 months-5.4 years) showed excellent cosmetic results and all the patients have remained completely dry. CONCLUSIONS: In patients with a classical picture of urinary incontinence caused by infrasphincteric ureteric ectopia associated with a small nonfunctioning kidney, video-laparoscopy, with its magnifying effect, can reliably confirm the diagnosis, locate the dysplastic kidney and allow its removal in the same setting. We propose that laparoscopy should be considered the investigation and treatment of choice in such patients, and should be undertaken without delay even if the dysplastic kidney or the ectopic ureteric orifice cannot be identified with all other conventional means. PMID- 10468731 TI - Low-dose desmopressin in the treatment of nocturnal urinary incontinence in the exstrophy-epispadias complex. AB - OBJECTIVE: To report our experience of the use of desmopressin to improve nocturnal dryness in patients who have undergone a staged reconstruction of the exstrophy-epispadias complex (EEC), who although continent by day, have nocturnal incontinence because their nocturnal urinary output exceeds their bladder capacity. PATIENTS AND METHODS: Seven children (aged 8-12 years) who had undergone a staged reconstruction for EEC (six with classical bladder exstrophy, one with incontinent epispadias) were treated with intranasal desmopressin for persistent nocturnal incontinence despite daytime dryness. Previous additional procedures for continence had been self-augmentation in one and periurethral collagen injection in three others. The criteria for inclusion in the study were: normal renal function, no upper tract deterioration, no urinary tract infections, spontaneous voiding during the day with dry intervals between micturitions, a postvoid residual volume of <10% of bladder capacity and night-time incontinence for 7 nights/week. Desmopressin was administered at bedtime at increasing dosages from 10 to 30 microg until effective. Body weight, arterial blood pressure, and serum electrolytes were measured, and all patients assessed using renal ultrasonography, a voiding diary and a nocturnal pad-test. RESULTS: Desmopressin at doses of 10-30 microg was successful in keeping all the patients dry. The nocturnal urinary output was decreased so that it did not exceed bladder capacity. There was only one minor side-effect (nose bleeding). CONCLUSIONS: In selected patients with EEC, desmopressin is effective in improving nocturnal dryness, with no significant side-effects. PMID- 10468732 TI - Antenatally detected pelvi-ureteric junction obstruction: concerns about conservative management. AB - OBJECTIVE: To review the conservative management of antenatally detected pelvi ureteric junction (PUJ) obstruction in children. PATIENTS AND METHODS: The study included 121 children (142 kidneys) with antenatally diagnosed PUJ obstruction managed over an 8-year period, including 21 who were affected bilaterally; the main focus of the study was on the 100 children affected unilaterally. The intent was to manage these patients conservatively, with immediate surgery (early pyeloplasty) only if renal function was initially compromised (renographic function < 40%). The indications for surgery subsequently (delayed pyeloplasty) included either a deterioration in renal function, the onset of symptoms or increasing hydronephrosis. RESULTS: The anteroposterior diameter and renal function were weakly and negatively correlated. The kidneys in the delayed pyeloplasty group had a greater initial mean anteroposterior diameter than had those in the early or the conservative groups (P<0. 05). The improvement in renal function was the lower in the delayed pyeloplasty group than in the early pyeloplasty group; the improvement was inversely related to the anteroposterior diameter. CONCLUSIONS: The conservative management of some antenatally detected PUJ obstructions probably results in greater loss of function, which is not recovered after surgery. PMID- 10468733 TI - Minimally invasive treatment of urinary tract calculi in children. AB - OBJECTIVE: To report experience of a broad multimodality approach to the treatment of calculi in children using extracorporeal shock wave lithotripsy (ESWL), ureteroscopy/laser lithotripsy, lithoclast and percutaneous nephrolithotomy (PCNL). PATIENTS AND METHODS: The treatment and outcome were reviewed in 43 children managed by a range of minimally invasive modalities, either singly or in combination, between 1990 and 1997. These patients represent a selected group deemed suitable for minimally invasive management during a period of developing experience with these techniques. Of this cohort, six children had previously undergone open stone surgery and contributory metabolic abnormalities were identified in seven. ESWL was the sole treatment modality in 24 children (56%). In five children (12%) ureteroscopy/laser lithotripsy was combined with ESWL, eight (18%) underwent ureteroscopy/laser lithotripsy alone, whilst three with bladder stones were treated with the lithoclast. Combined therapy including PCNL was required in three patients. RESULTS: Of the 43 children treated, 38 (88%) were rendered stone-free. Metabolic disorders accounted for three of the five cases of residual calculi. Complications requiring intervention occurred in two children (7%) and three subsequently underwent open pyelolithotomy or ureterolithotomy after unsuccessful minimally invasive treatment. CONCLUSIONS: Used selectively, the range of minimally invasive procedures available for adults, including ureteroscopy and PCNL, can be safely and effectively extended to the treatment of urinary tract calculi in children. The role of open surgery will diminish further with the availability of specialized instruments for paediatric PCNL. PMID- 10468734 TI - Carbachol-induced sustained tonic contraction of rat detrusor muscle. AB - OBJECTIVE: To investigate the underlying contractile mechanism of the sustained tonic contraction (SuTC) induced by repetitive carbachol application in rat detrusor muscles. MATERIALS AND METHODS: Longitudinal muscle strips with no mucosa were obtained from the anterior wall of the urinary bladder in 12-week-old Sprague-Dawley rats. Carbachol (5 micromol/L) was applied repetitively to induce SuTC. The carbachol-induced SuTC was assessed in the presence of various Ca2+ channel blockers and drugs affecting intracellular Ca2+ concentration. RESULTS: The first application of carbachol elicited a large phasic contraction followed by a tonic contraction (TC); the carbachol-induced contraction was completely reversed by washing out the solution. However, the initial phasic contraction was not reproduced after a second or further application of carbachol. There was consistently only a SuTC with no phasic contraction. The amplitude of the SuTC was 85% of the TC induced by the first carbachol application. The application of atropine (1 micromol/L) to the bath completely blocked SuTC. The carbachol induced SuTC was insensitive to nicardipine (5 micromol/L) and extracellular polyvalent cations (1 mmol/L, La3+, Co2+, Cd2+, Ni2+ ). Moreover, a similar SuTC was induced even after the complete elimination of extracellular Ca2+ by adding 2 mmol/L EGTA to the Ca2+-free Tyrode solution. To exclude intracellular Ca2+ sources related to the sarcoplasmic reticulum (SR), the effects of SR Ca2+ pump inhibitors, cyclopiazonic acid (CPA, 10 micromol/L) and thapsigargin (0.5 micromol/L) were tested. The carbachol-induced SuTC was insensitive to pretreatment with CPA and/or thapsigargin. To deplete the ryanodine-sensitive Ca2+ pool, muscle strips were repetitively stimulated with caffeine (10 mmol/L) in the presence of 10 micromol/L ryanodine, which did not affect the carbachol induced SuTC. CONCLUSIONS: Although the characteristics of the carbachol-induced SuTC have not been defined, these results show that a significant proportion of the carbachol-induced contraction in rats is contributed by the SuTC, which is present even in the complete absence of external Ca2+. The SuTC was not affected by limiting the contributions of internal Ca2+ sources. This suggests that the SuTC in rat bladders is unrelated to known Ca2+ mobilization mechanisms. PMID- 10468735 TI - MUC1 mucin as a tumour marker in bladder cancer. AB - OBJECTIVE: To evaluate serum MUC1 levels (a high molecular weight glycoprotein which is upregulated and abnormally glycosylated in bladder cancer and other carcinomas) in patients with a variety of stages and grades of transitional cell carcinoma (TCC) of the bladder, to assess its potential as a tumour marker. PATIENTS AND METHODS: Blood samples were taken before treatment in 87 patients with TCC of the bladder and in 31 controls undergoing cystoscopy for benign conditions. Serum MUC1 levels were estimated with an enzyme-linked immunosorbent assay using the C595 monoclonal antibody. RESULTS: Of patients with T4 tumours, 47% had MUC1 levels above the normal range (P<0.001); patients with T3 tumours also had significantly higher MUC1 levels than controls. The overall sensitivity was only 24% for all tumours when the upper limit of normal was defined as 4.8 U/mL; the specificity was 97%. CONCLUSION: Serum MUC1 is not as useful tumour marker for screening, as it has a low sensitivity. However, MUC1 levels are high in advanced disease and serum MUC1 levels may be useful for disease monitoring. PMID- 10468736 TI - Does maternal stress induce abnormal descent of the testis in prepubertal rats? AB - OBJECTIVE: To investigate whether prenatal maternal stress, considered to alter plasma testosterone concentration and induce a lack of testosterone surge in male rat fetuses, has any effect on the growth of the processus vaginalis and testicular descent in male offspring. MATERIALS AND METHODS: Pregnant rats were divided into two groups. In group 1, the rats were placed three times daily for 60 min each session in Plexiglas rat holders (13x6x8 cm) illuminated by two 150-W flood lights from day 14 to day 18 of gestation during the dark phase of the animals' light (12 h) and dark (12 h) cycle. In group 2, pregnant females were not handled and acted as controls. After birth, the anogenital distance of both the male and female rats was measured, and the length of the processus vaginalis was measured at 7 days of age in some rats. Thereafter, at 21, 24, 27 and 30 days of age, testicular descent was assessed in the remaining male rats. RESULTS: In group 1, at 21, 24, 27 days of age, testicular descent was delayed in the stressed rats compared with the control rats. At 30 days of age, five of 36 stressed male offspring showed abnormal testicular descent; three testes were in the superficial inguinal position and two were above the scrotum. Both the anogenital distance and the length of the processus vaginalis were significantly less in the stressed male rats than in the controls. CONCLUSIONS: Maternal stress induced a female-like anogenital distance in male rat fetuses, and caused delayed and abnormal descent of the testis, by inhibiting the growth of the processus vaginalis after birth. Prenatal stress may therefore be involved in the delay of testicular descent in neonatal rats by inducing a mild anti-androgenic effect in the fetus. PMID- 10468737 TI - The evaluation of sympathetic system-related contractile activity of the rat vas deferens after ligation and intra-abdominal placement of the testis. AB - OBJECTIVE: To evaluate the contractile response of the vas deferens in a model of stress, to determine any changes in sympathetic activity as a result of stress in the ipsilateral testis, which decreases blood flow to the contralateral testis. MATERIALS AND METHODS: The study comprised two groups of six rats each; group 1 underwent a sham operation, and in group 2 the right testis was placed into the abdominal cavity and the vas deferens ligated. After 30 days, the vasa deferentia were resected bilaterally and their isometric contractions recorded. Electrical field stimulation (EFS) was applied through a pair of platinum electrodes and concentration-response curves constructed for noradrenaline at 37 degrees C and to a solution containing 80 mmol/L K+. RESULTS: The vasa deferentia in both groups showed similar contractile responses to EFS, which were frequency dependent and maximal at 80 Hz. Noradrenaline-induced contractile activity was lower in amplitude in the vasa deferentia of group 2 than in the contralateral and ipsilateral vasa deferentia of group 1, which were not significantly different from each other. All groups responded similarly to high K+. CONCLUSION: Intra-abdominal placement of the testes with vas deferens ligation decreased the contractile response to noradrenaline in the ipsilateral vas deferens without altering the contractile response to EFS and high K+. This difference could be caused by a reduction in the number of postjunctional alpha-adrenergic receptors or decreased receptor sensitivity. Both possibilities suggest that the vas deferens may initiate sympathetic activity, which may be responsible for contralateral testicular deterioration. PMID- 10468738 TI - Point of technique: Management of penile and scrotal elephantiasis. PMID- 10468739 TI - Point of technique: The retrograde Add-a-Cath technique: an aid to changing the difficult suprapubic catheter. PMID- 10468740 TI - Point of technique: Ureterocalycostomy in the management of pelvi-ureteric junction obstruction in the horseshoe kidney. PMID- 10468741 TI - Rupture of the ureter: an unexpected complication of microdiskectomy. PMID- 10468742 TI - Ureteric obstruction after lumbar diskectomy. PMID- 10468743 TI - Coccidioidomycosis prostatitis associated with prostate cancer. PMID- 10468744 TI - Paternity in a patient with seminoma and carcinoma in situ in a solitary testis treated by partial orchidectomy. PMID- 10468745 TI - Moderate preputial dysplasia in a young child. PMID- 10468746 TI - Body piercing: an unusual cause of priapism. PMID- 10468747 TI - Transitional cell carcinoma of the bladder in a patient on clean intermittent catheterization. PMID- 10468748 TI - Renal adenomatosis and unilateral hydronephrosis. PMID- 10468749 TI - Sertoli tumour presenting with stress incontinence in a patient with testicular feminization. PMID- 10468750 TI - Vaginal bleeding as a presentation of metastatic renal cell carcinoma. PMID- 10468751 TI - A prospective study of nonmedical prophylaxis after a first kidney stone. AB - OBJECTIVE: To assess, in a multicentre prospective randomized study, the effectiveness of specific nonmedical prophylaxis and nonspecific dietary prophylaxis in patients treated after experiencing their first idiopathic calcium containing kidney stone. PATIENTS AND METHODS: From 1991 to 1994, 242 patients with idiopathic calcium urinary stones from three urological centres were randomly assigned into two groups. In group 1 (intervention, 113 patients), a specific dietary regimen was recommended and thereafter corrected according to a comprehensive metabolic evaluation. Group 2 (control, 94 patients) underwent only minimal metabolic screening and used general dietary measures, with no regular follow-up. An increased fluid intake was recommended in both groups. After 3 years, the effectiveness of the prophylactic and dietary regimens was evaluated in 207 patients. RESULTS: At entry, the clinical characteristics were comparable in the two groups, with only extracorporeal lithotripsy being more frequent in group 2. Almost 80% of patients reported a high intake of meat and a low intake of dairy products. In group 1 and 2, a recurrent stone was encountered in seven (6%) and 18 (19%) patients, respectively; recurrent and growing stones were found in eight (8%) and 22 (23%) patients, respectively (P<0. 01). There was a higher incidence of bilateral residual (or untreated) stones, and Weddellite stones (P<0.001) among patients with a recurrent or growing stone. An analysis of the dynamics of the metabolic variables indicated the significance of repeated dietary counselling. CONCLUSION: Specific dietary therapy, adjusted according to a metabolic evaluation, is more effective than nonspecific general dietary recommendations in preventing the formation of a second urinary stone. PMID- 10468752 TI - Preliminary results of the alternating administration of natural interferon-alpha and recombinant interferon-gamma for metastatic renal cell carcinoma. AB - OBJECTIVE: To evaluate the efficacy and toxicity of the alternating administration of natural (n) interferon (IFN)-alpha and recombinant (r) IFN gamma for metastatic RCC. PATIENTS AND METHODS: The study comprised 24 patients (median age 60 years, range 42-77), 20 of whom were evaluable for response and all 24 evaluable for toxicity. Initially, nIFN-alpha was administered subcutaneously on days 1 and 3, and rIFN-gamma on day 2, for 1-2 weeks in the evening or at night, both at doses of 3 MU. If this regimen was tolerated, nIFN alpha and rIFN-gamma were administered at the same doses on days 1, 3 and 5, and on days 2 and 4, respectively. RESULTS: There were three complete remissions and two partial remissions, giving a total response rate of 25%. All responders (complete plus partial remission) had undergone nephrectomy. Multiple lung metastases completely disappeared from four responders. The median and maximum time to remission in the responders were 2 and 7 months, respectively. The survival time of the responders was significantly longer than that of those not responding (stable and progressive disease, P=0.0202). Toxicities were mostly limited to WHO grades 1 and 2, with grade 3 leucopenia and grade 4 hepatic dysfunction in only one patient each. These toxicities were transient and there were no treatment-related deaths. CONCLUSION: The alternating administration of nIFN-alpha and rIFN-gamma is an effective treatment for metastatic RCC. This treatment is particularly suitable for patients who have undergone nephrectomy and have lung metastases. PMID- 10468753 TI - Follow-up guidelines for nonmetastatic renal cell carcinoma based on the occurrence of metastases after radical nephrectomy. AB - OBJECTIVE: To define guidelines for the follow-up management of nonmetastatic renal cell carcinoma (RCC), by assessing tumour recurrences and the clinical course in patients who had undergone radical nephrectomy. PATIENTS AND METHODS: The records of 187 patients with pT1-3, N0-X, M0 RCC who underwent radical nephrectomy between 1982 and 1997 were reviewed prospectively. Clinicopathological variables were compared with the time of first recurrence, site of metastasis and reason for diagnosis. RESULTS: Metastases were diagnosed in 98 sites in 56 of the 187 patients (30%). The risk for developing metastases increased with stage; 80% of the patients had their metastases diagnosed within 3 years (median 14.5 months) after nephrectomy. The time to first diagnosis was longer for patients with pT1 tumours and for those with skeletal metastases. The cause-specific 5-year survival rate for pT1 tumours was 95%, for pT2 87% and for pT3 tumours 37%. All patients with diploid pT1-2 RCC survived, having a survival advantage over those with aneuploid pT1-2 tumours (P=0.018). Also, pT1-2 tumours of < 5 cm were associated with better survival rates. Among 74 patients with pT3 tumours, 45 got metastases; DNA ploidy in these tumours did not influence survival. Of 30 patients with lung metastases, 28 were diagnosed during follow up, while 25 of 26 other metastatic sites were diagnosed because of symptoms. CONCLUSIONS: The risk for tumour progression depends mainly on stage; these results indicate no need for follow-up in patients with diploid pT1-2 tumours or with aneuploid pT1 tumours of < 5 cm. For patients with aneuploid pT1-2 tumours of > 5 cm and pT3 tumours, follow-up is indicated. PMID- 10468754 TI - The genetics of transitional cell carcinoma: progress and potential clinical application. PMID- 10468755 TI - Elevated urinary fibronectin levels after transurethral resection of bladder tumour: a possible role in patients failing therapy with bacillus Calmette Guerin. AB - OBJECTIVE: To investigate fibronectin levels in urine samples from patients with noninvasive transitional cell carcinoma (TCC) of the bladder immediately and for 4 weeks after transurethral resection of bladder tumour (TURBT), to determine whether soluble fibronectin within the bladder, which blocks the attachment of bacillus Calmette-Guerin (BCG), might lower the efficacy of BCG therapy over this period. PATIENTS AND METHODS: Urinary fibronectin was measured using an enzyme linked immunosorbent assay in 25 patients with superficial bladder TCC who underwent TURBT for complete resection. Eight samples were collected for each patient, one before and seven during the 4 weeks after TURBT. RESULTS: High levels of urinary fibronectin were detected in 18 patients (72%) after TURBT. In 16 patients the fibronectin level returned to normal within 2 weeks of surgery. The other two patients showed elevated levels of fibronectin for > 4 weeks. CONCLUSIONS: These results show that urinary fibronectin concentration is significantly increased in most patients after TURBT and this should be considered in patients who receive BCG therapy. Treatment within the first 2 weeks after TURBT may be associated with a high failure rate, as urinary fibronectin levels were increased significantly in about three-quarters of these patients during that period. Indeed, the persistent elevation of fibronectin, occurring in two of the present patients, may be responsible for some of the failures of BCG therapy when it is administered 2-5 weeks after surgery. PMID- 10468756 TI - Does the position of the patient affect the presence and/or degree of reflux in an ileal conduit urinary diversion? AB - OBJECTIVE: To assess the effect of patient position (supine, sitting or standing) on ileo-ureteric reflux in patients with an ileal conduit urinary diversion, in whom such reflux is normally detected when they are supine during a retrograde loopogram. PATIENTS AND METHODS: The study included 10 patients with an ileal conduit as a primary urinary diversion; a loopogram was obtained with the patient upright or supine and a further film taken with the patient supine but at 45 degrees to the ground. RESULTS: When supine, free ileo-ureteric reflux occurred into both ureterorenal units in eight patients. The remaining two patients, who had previously undergone unilateral nephrectomy, also had reflux into their existing renal units. Of the 18 units, 15 had grade III and three had grade IV reflux. In the upright and 45 degrees position, reflux still occurred in al ureterorenal units. The patient's position did not affect the degree of reflux in 16 units, but in one unit with grade IV reflux and another with grade III reflux, the reflux was one grade less severe. CONCLUSIONS: Ileo-ureteric reflux is common after ileal conduit diversion and may contribute to the likelihood of renal deterioration. The presence and/or degree of reflux is generally not affected by the position of the patient. PMID- 10468757 TI - The Mitrofanoff urinary diversion for complex vesicovaginal fistulae: experience from Uganda. AB - OBJECTIVE: To evaluate the results of Mitrofanoff continent urinary diversion in a group of women with persistent severe incontinence after vesicovaginal fistula (VVF) secondary to obstructed labour. PATIENTS AND METHODS: Seven women with severe incontinence following a VVF were offered the Mitrofanoff procedure after all other attempts had failed to restore continence. In three patients a caecocystoplasty formed the urinary reservoir and in four the bladder was used. In all seven patients the appendix was used as the conduit for self catheterization. The mean (range) follow-up was 10 (3-14) months. RESULTS: One patient died postoperatively from hepatic failure, which could not be attributed to the particular procedure. One patient required re-operation at 10 days to adjust the conduit, but of the six patients who recovered, all are fully continent and self-catheterizing with no difficulty. CONCLUSION: The Mitrofanoff procedure appears to be a valuable technique to restore continence in this difficult group of patients. PMID- 10468759 TI - A comparative study of the effects of a colorectal valve on the oro-anal transit time in patients with an augmented and valved rectum for urinary diversion. AB - OBJECTIVES: To evaluate the effect of the interposed colorectal valve on the flow of the proximal colonic contents to the rectum in patients with a valved urinary diversion to the rectum. PATIENTS AND METHODS: The oro-anal transit time (OATT) and segmental colonic transit time (SCTT) were estimated in two groups of 15 patients each (10 women and five men). In group 1, patients had a colorectal valve and in group 2 (control) they had no valve in the rectosigmoid area. The OATT and SCTT were evaluated using the 7-day method, whereby each patient swallowed 10 ring-shaped radio-opaque markers daily for 6 days, and a single abdominal radiograph was taken on day 7. The OATT and SCTT were then estimated as the number of retained markers divided by the daily dose of markers. RESULTS: The mean (sd) OATT was not significantly different (P=0.185), at 1.85 (1.21) days in group 1 and 2.49 (1.38) days in group 2. There were no sex differences in the values between the groups. The SCTT in the four areas of the colon again showed no significant differences within or between groups (P>0.05). CONCLUSION: Using a colorectal valve to functionally isolate the urinary rectal reservoir and thus reduce the high incidence of metabolic acidosis has no significant effect on intestinal transit times. PMID- 10468758 TI - A prospective randomized study comparing absorbable and nonabsorbable staples in constructing antireflux valves of urethral hemi-Kock pouches. AB - OBJECTIVE: To determine the long-term complications associated with constructing an antireflux nipple valve using absorbable or nonabsorbable staples in the urethral hemi-Kock pouch. PATIENTS AND METHODS: Fifty patients (all men, median age 47 years, range 28-73) undergoing construction of a urethral hemi-Kock pouch were randomized prospectively into two equal groups. In group 1, absorbable (polyglyconate) staples were used for the construction, and in group 2 nonabsorbable (metal) staples were used but omitting the staples applied at the tip of nipple valve. Assessment during the follow-up of 6-88 months comprised urine analysis, culture and sensitivity, measurement of serum creatinine, electrolytes and blood gases, and plain abdominal radiography, intravenous urography and micturating pouchography. RESULTS: In group 1, 21 patients were evaluable; a pouch stone was detected in one, extussusception of the valve in one and valve stenosis in one. In group 2, 20 patients were evaluable; pouch stones were detected in six, extussusception of the valve in three and valve stenosis in one. CONCLUSION: Compared with nonabsorbable (metal) staples, the use of absorbable (polyglyconate) staples significantly decreased the incidence of pouch stone formation and improved valve stability in patients with a urethral hemi Kock pouch. PMID- 10468760 TI - The tolerability of urodynamic studies and flexible cysto-urethroscopy used in the assessment of men with lower urinary tract symptoms. AB - OBJECTIVE: To determine the overall tolerability of urodynamic studies used in the assessment of men with lower urinary tract symptoms (LUTS), by assessing the objective and subjective morbidity experienced during and after urodynamic studies, and to assess the voiding complaints caused by the combination of urodynamic studies with flexible cysto-urethroscopy. PATIENTS AND METHODS: A total of 103 men with LUTS, who underwent a urodynamic study combined with flexible cysto-urethroscopy, completed a questionnaire designed to assess objective and subjective symptoms and degree of bother, with emphasis on the urodynamic study. In addition, a urine specimen was analysed and cultured. In all, 78 patients who underwent a second urodynamic study completed the questionnaire twice. RESULTS: The results of the first questionnaire showed that more than half of the patients experienced some urge after the urodynamic study and cysto-urethroscopy (56%); 35% of the patients experienced little and 19% experienced severe voiding discomfort after the combined investigations, compared with 24% and 5%, respectively, after a urodynamic study alone. Three patients (3%) had a symptomatic urinary tract infection. Haematuria, increased voiding frequency and increased nocturia occurred occasionally. Most of the patients found the urodynamic study less bothersome than they had expected (64%) and only 9% found it worse than expected. The overall degree of discomfort, experienced during and after the urodynamic study combined with cysto-urethroscopy, was low, and after a second urodynamic study was even lower. CONCLUSION: In contrast with earlier results, this clinic-based urodynamic investigation was associated with a low proportion of urinary tract infection, and low objective and subjective morbidity. The combination of a urodynamic study with a flexible cysto urethroscopy does not cause significant additional voiding complaints. Most patients find urodynamic studies tolerable and not very bothersome. PMID- 10468761 TI - A new automated system for urine analysis: a simple, cost-effective and reliable method for distinguishing between glomerular and nonglomerular sources of haematuria. AB - OBJECTIVE: To determine the ability, reliability and accuracy of a new automated system of urine analysis in differentiating glomerular from nonglomerular bleeding in the initial investigation of haematuria, and compare its efficacy with conventional phase-contrast microscopy (PCM). PATIENTS AND METHODS: One hundred and six urine samples from patients in whom the final diagnosis was available were analysed using electrical flow impedance to detect, enumerate and size red blood cells in a conductive fluid (the cellfacts analyser, Microbial Systems Ltd, Coventry, UK). All the samples were also tested using a dipstick method and PCM was carried out for comparison on 45 of the 106 urine specimens. The results of cellfacts analysis were correlated with the final diagnoses to assess sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of this method; the costs were also analysed. RESULTS: Sixty-nine urine samples tested positive for blood on dipstick urine analysis and all these were confirmed to have red blood cells on cellfacts analysis. The remaining 37 samples were negative for blood on dipstick testing and cellfacts analysis, although seven patients had been referred with previously detected microscopic haematuria, none of whom were found to have any detectable pathology in the urinary tract on clinical examination and investigations. The remaining 30 patients were diagnosed to have urological or nephrological conditions with no haematuria. In the positive group, 20 (29%) patients were from the glomerular group, with a mean (range) red blood cell size of 4.25 (4-5.1) micrometer, and 49 (71%) from the nonglomerular group, with red blood cells of 5.47 (4.67-5.70) micrometer. These ranges overlapped at 4.67-5.1 micrometer at the decision threshold of 4.75 micrometer, the distribution of dysmorphic and eumorphic red blood cells for the glomerular group was 18 (90%) and two (10%), respectively, and for the nonglomerular group was 2 (4%) and 47 (96%), respectively. The sensitivity, specificity, PPV and NPV were 90%, 96%, 90% and 96%, respectively. Consumable and labour costs were very low. CONCLUSIONS: Cellfacts analysis is a simple, rapid, objective and cost-effective method for differentiating glomerular from nonglomerular urinary red blood cells, especially when few such cells are present. PMID- 10468762 TI - The urodynamic profile of prostatodynia. AB - OBJECTIVE: To evaluate urodynamically patients with prostatodynia, and thus define a specific urodynamic pattern that might add to the pathophysiology of the syndrome and possibly aid definitive treatment. PATIENTS AND METHODS: Forty-three patients (mean age 38.1 years, sd 9.25, range 24-59) with symptoms suggestive of chronic prostatitis, e.g. dysuria, frequency of micturition and a burning sensation in the perineum, were classified as having prostatodynia after excluding prostatic infection by standard bacteriological methods. Thereafter, the patients were evaluated urodynamically, including the measurement of free flow-rate, filling (water) cystometry, a pressure/flow study of micturition, with fluoroscopy and electromyography of the external urethral sphincter. RESULTS: Twenty-eight patients (65%) had a low maximum free flow rate (Qmax ), with a mean (sd) of 10.91 (1.26) mL/s. Of the 42 patients who underwent filling cystometry, 26 (62%) had a first sensation of filling, and 28 (67%) a first desire to void, at low volumes (<150 mL and <300 mL, respectively). Of the 25 patients who underwent a pressure/flow study, 16 (64%) had an obstructive pattern of micturition, as defined by a low Qmax of 10.04 (1.38) mL/s and a high intravesical pressure at Qmax of 83.3 (5.3) cmH2O. The site of obstruction was at the level of the bladder neck, as confirmed by fluoroscopy. CONCLUSION: A significant proportion of these patients had a particular urodynamic pattern of functional infravesical obstruction at the level of the bladder neck and sensory disturbances. PMID- 10468763 TI - Malacoplakia: von Hansemann's disease. PMID- 10468764 TI - Prostatic involvement in men with febrile urinary tract infection as measured by serum prostate-specific antigen and transrectal ultrasonography. AB - OBJECTIVE: To determine the frequency of prostatic involvement in men with community-acquired febrile urinary tract infection. PATIENTS AND METHODS: This prospective study included 70 men (18-85 years old) who had a fever of >/=38.0 degrees C, symptoms or signs of urinary tract infection and a positive urine culture. Serum prostate-specific antigen (PSA) was measured and transrectal ultrasonography of the prostate and seminal vesicles performed during the acute phase of the disease and during a 1-year follow-up. RESULTS: Although only six patients had a tender prostate on digital rectal examination, the initial serum PSA level was elevated in 58 (83%) patients (median 14 ng/mL, range 0.54-140). There was no correlation between PSA levels, patient age, inflammatory response to infection or presence of positive blood cultures. Despite a rapid decline in PSA level after one month, there was a protracted decrease in some patients. After 3 months the median prostate volume was reduced by 31% (range 11-54; P<0.001) in 46 of 55 patients examined, and the width of the right and left seminal vesicle was reduced by 14% and 22%, respectively. The reductions in PSA and prostate volume were significantly correlated (r=0.36, 95% confidence interval 0.09-0.58; P=0.01). CONCLUSION: These results show that the prostate and seminal vesicles are frequently involved in men with febrile urinary tract infection and that PSA may be a useful marker of prostatic infection. The slow decline of PSA levels in some patients after appropriate antibiotic treatment indicates a protracted healing process and should be considered when PSA is used to detect prostate cancer. PMID- 10468765 TI - On the chequered history of vasectomy. PMID- 10468766 TI - Is the measurement of serum testosterone routinely indicated in men with erectile dysfunction? AB - OBJECTIVE: To investigate the usefulness of serum testosterone levels as a relevant or useful indicator of sexual potency in men attending an erectile dysfunction clinic. PATIENTS AND METHODS: Ninety consecutive men attending the erectile dysfunction clinic completed a sexual-activity questionnaire, and underwent a focused physical examination and questioning about their medical history. The serum testosterone level was measured in all patients and the results analysed in relation to the patient's age. Patients with low serum testosterone levels commenced replacement therapy comprising three intramuscular injections of testosterone (Sustanontrade mark, Organon, The Netherlands) 250 mg every third week. Potency status and serum testosterone were reassessed after 3 months' treatment. RESULTS: Of the 90 men, 28 (31%) were aged < 50 years whilst 62 (69%) were >/=50 years old. Nineteen (21%) patients overall had low testosterone levels; four of these were < 50 and 15 were >/=50 years old. Five of 90 patients had a decreased libido; two of these also had low testosterone levels and all were < 50 years old. Testosterone levels returned to normal in all patients who received replacement therapy but potency returned in only two (10%); both were in the older group. CONCLUSION: Measuring testosterone was not helpful in assessing potency or libido and low serum levels were not related to age. Correcting low testosterone did not improve either impotence or libido. PMID- 10468767 TI - The role of fine-needle aspiration cytology of the testis in the diagnostic evaluation of infertility. AB - OBJECTIVE: To compare the diagnostic value of cytological smears of fine-needle aspirate with that of histological sections obtained by testicular biopsy in the evaluation of infertility. PATIENTS AND METHODS: The study included 60 patients with azoospermia or severe oligospermia (sperm counts of < 5 million/mL). A detailed history was taken and the external genitalia examined. Testicular samples were obtained by fine-needle aspiration using a 10 mL syringe and 21 G needle; the air-dried smears were stained using the May-Grunwald-Giemsa method. A testicular biopsy was taken after exploring the same tract and these sections were stained with haematoxylin and eosin. The cytological smears and histological sections were examined separately and the results compared. RESULTS: The cytological smears were divided into three groups: normal, with all spermatogenic cells present; Sertoli-cell only syndrome; and spermatogenic arrest (all spermatogenic cells present except spermatids and spermatozoa). There was a good correlation between the diagnosis on the cytological smears and histological sections in 58 of 60 patients (97%). In two patients there was scanty aspirate and few cells, giving an inaccurate diagnosis. Interstitial cells were not detected on cytological smears. In three patients, Sertoli-cell only was diagnosed on cytology, and histology revealed additional features of Leydig cell hyperplasia. CONCLUSION: Fine-needle aspiration of the testis is a simple and minimally invasive technique for diagnostic evaluation, especially in patients with obstructive azoospermia. Patients with abnormal findings on fine-needle aspiration cytology may be evaluated histologically by further biopsy. PMID- 10468768 TI - Ischaemic preconditioning protects the rat kidney from reperfusion injury. AB - OBJECTIVE: To examine the possible role of ischaemic preconditioning (IPC), an adaptive pathophysiological phenomenon that increases tolerance to ischaemia reperfusion (I-R) injury, in renal protection when rats are presented with an I-R challenge. MATERIALS AND METHODS: Female Wistar rats (n=36) were divided randomly into four groups: (A) sham-operated controls; (B) IPC only; (C) renal ischaemia (RI) only; and (D) IPC+RI. The left kidney in groups B and D was preconditioned with four cycles of renal artery occlusion lasting 4 min, each occlusion separated by 11 min of reperfusion. The ischaemic insult, applied in groups C and D, comprised 40 min of sustained left renal artery occlusion. In Group D, the IPC cycle was completed 5 min before the start of the ischaemic insult. Differential left renal function was calculated by 99mTc-labelled dimercaptosuccinic acid scintigraphy at 0, 2 and 9 days after treatment, and expressed as a percentage of the total renal uptake. RESULTS: The mean (sem) maximum decrease in left renal function, to 14.5 (4.3)% of the total, occurred on day 2 in Group C. The equivalent value in Group D showed relative preservation of function, at 36.0 (3.5)% (P=0.001 compared with Group C). The mean left renal function improved by day 9, to 39.6 (6.7)% (Group C) and 48.6 (1.5)% (Group D). The mean left renal function in Group B (50.5-53.9%) did not differ from that in controls (49.4 51.4%). CONCLUSION: An IPC regimen applied 5 min before RI in the rat significantly protects it from the functional impairment associated with ischaemia and reperfusion. PMID- 10468769 TI - Expression of p75(LNGFR) and Trk neurotrophin receptors in normal and neoplastic human prostate. AB - OBJECTIVE: To analyse the occurrence and cell distribution of p75(LNGFR) and Trk neurotrophin receptors in normal prostate, benign prostatic hypertrophy (BPH) and prostate carcinoma, and to determine the effect of androgen suppression on the expression of these proteins in prostate cancer samples. MATERIALS AND METHODS: The study comprised formalin-fixed and paraffin-embedded material, obtained during surgery and from cadavers during removal of organs for transplantation. Light microscopy immunohistochemistry was carried out using polyclonal antibodies against Trks, and a monoclonal antibody against p75(LNGFR). General markers for epithelial and endocrine cells were assessed in parallel. RESULTS: TrkA immunoreactivity (IR) was restricted to the basal epithelial cells in some acini (37%). This pattern remained unchanged or IR extended to the whole acini in BPH, and varied widely in prostate cancer. In normal tissue and BPH, TrkC IR was detected exclusively in the stroma. Nevertheless, it progressively increased in the epithelial cells of well-differentiated to moderately differentiated prostate carcinoma, whereas in stromal cells there were no substantial changes. TrkB IR was absent in all the samples. There was weak p75(LNGFR) IR in normal epithelial cells, which increased in prostate cancer and to a lesser extent in BPH. Androgen suppression was ineffective in reversing TrkA modifications, whereas it caused a decrease in the expression of TrkC and p75(LNGFR). CONCLUSION: The abnormal growth of prostatic epithelium is accompanied by increased TrkA expression and the induction of TrkC expression in epithelial cells. These results suggest that neurotrophins could be involved in the abnormal growth of the human prostate, acting through specific Trk signal-transducing receptors whose expression is regulated by androgens. PMID- 10468770 TI - An in vivo evaluation of the therapeutic potential of sympatholytic agents on premature ejaculation. AB - OBJECTIVE: To evaluate the therapeutic potential of sympatholytic agents on premature ejaculation in an animal model, using monitoring of rat seminal vesicle pressure change in response to electrical stimulation of the lesser splanchnic nerve. MATERIALS AND METHODS: Male Wistar rats (aged 12-14 weeks) were injected intra-arterially with sympatholytic agents (phenoxybenzamine, prazosin, WB-4101, chloroethylclomidine, yohimbine and RX 821002) at various concentrations 10 min before electrical stimulation of the lesser splanchnic nerve. The change in phasic tension (triangle upmmHg) of the seminal vesicle induced by electrical nerve stimulation before and after the addition of sympatholytic agents was used for statistical analysis. The maximum inhibition and the concentration required to induce 50% inhibition of the maximal contractile response (IC50) were obtained from the concentration-response curves, and used to determine the potency of test agents. RESULTS: The seminal vesicle contractile response to electrical nerve stimulation was suppressed in a dose-dependent manner by all test drugs except RX 821002. The mean (sd) maximal inhibition was 78.4 (9. 3)% by 0.03 mg/kg of phenoxybenzamine, 77.1 (10.1)% by 0.03 mg/kg of WB-4101, 73.4 (6.0)% by 0.1 mg/kg of yohimbine, 67.9 (9.7)% by 0.1 mg/kg of prazosin, 75.5 (7.5)% by 3 mg/kg of chloroethylclomidine and 25.3 (4.8)% by 0.01 mg/kg of RX 821002. The potencies of WB-4101 (IC50 3 microgram/kg) and yohimbine (IC50 0.8 microgram/kg) were similar to that of phenoxybenzamine (IC50 0.5 microgram/kg) and much higher than that of prazosin (IC50 0.03 mg/kg) or chloroethylclomidine (IC50 0.3 mg/kg). CONCLUSIONS: Phenoxybenzamine, prazosin, WB-4101, chloroethylclomidine and yohimbine all inhibit the contractile response of the rat seminal vesicle to electrical nerve stimulation. As phenoxybenzamine is effective in treating premature ejaculation, the comparable in vivo potencies of WB-4101 and yohimbine strongly suggest that they have clinical therapeutic potential. PMID- 10468771 TI - The long-term culture of porcine urothelial cells and induction of urothelial stratification. AB - OBJECTIVE: To assess porcine urothelial cell cultures and the in vitro induction of urothelial stratification in long-term cultures, to study their morphological, functional and genetic behaviour, and thus provide potential autologous urothelium for tissue-engineered substitutes for demucosalized gastric or colonic tissue. MATERIALS AND METHODS: Primary cultures of porcine urothelium were established and the cells passaged thereafter. Cell specificity was confirmed by cytokeratin analysis, cell membrane stability assessed using lactate dehydrogenase leakage, cell de-differentiation by gamma-glutamyl transferase activity and genomic stability by karyotype investigations. Histology and scanning electron microscopy were performed to study the cultured cells and the stratified constructs. Furthermore, collagen matrices were tested as cell scaffolds. RESULTS: The cells were cultured for 180 days; 10 subcultures were established during this period. Stratification was induced in a culture flask and on a collagen matrix. Cytokeratins 7, 8, 17 and 18 were expressed in all cultures, and cell membranes were stable, with no evident de-differentiation. The cultures were stable in their genotype and no chromosomal aberrations were found. The histology and immunohistochemistry of the stratified porcine constructs, and cell membrane stability and cell de-differentiation, were compared with those in the human system. CONCLUSION: Pig and human urothelial cells can be cultured over a long period with no signs of senescence. Urothelial stratification can be induced in vitro. The collagen matrix seems to be an excellent scaffold, allowing cell adherence and growth. PMID- 10468772 TI - Contractile responses to alpha1-adrenoceptor agonists in isolated human male and female urethra. AB - OBJECTIVE: To investigate the contractile responses mediated through alpha1 adrenoceptors in human urethra and to evaluate the effectiveness of NS-49 [(R)-( )-3'-(2-amino-1-hydroxyethyl)-4'-fluoromethanesulphonanilide++ + hydrochloride], a novel alpha1-adrenoceptor agonist, against contraction of the human urethra. MATERIALS AND METHODS: The contractile responses were assessed in 10 male prostatic urethrae and six female urethrae. Antagonism was evaluated in the urethra using phenylephrine, a nonselective alpha1-adrenoceptor agonist, cumulatively applied > 20 min after applying 0.1 micromol/L prazosin or 0.1 micromol/L 5-methylurapidil, a selective alpha1A-adrenoceptor antagonist. Agonism was determined in both male and female urethrae to obtain the concentration response curve for the agonist. RESULTS: Phenylephrine caused both male and female urethrae to contract, and showed high potency and efficacy. Prazosin antagonized these contractions with low affinity (apparent pKB of 8.30 in male urethrae). 5-Methylurapidil, also antagonized the contractions with low affinity (apparent pKB of 7.88 in male urethrae). Noradrenaline and phenylephrine caused both male and female urethrae to contract, with high potency and efficacy. A novel and selective alpha1A-and alpha1L-adrenoceptor agonist, NS-49, induced contractile responses with high potency and moderate efficacy, whereas methoxamine induced contractions with low potency and moderate efficacy. Norephedrine was a very weak contractile agonist. CONCLUSION: In the human urethra, phenylephrine-induced contractions were mediated through alpha1L adrenoceptors and not through alpha1A-adrenoceptors. Contractions of the human urethra induced by NS-49 were also mediated mainly through alpha1L-adrenoceptors, with high potency and moderate efficacy. NS-49 may therefore be useful for the treatment of urinary stress incontinence, with minimal side-effects because it has subtype selectivity. PMID- 10468773 TI - Transrectal ultrasonography-guided echo-enhanced seminal vesiculography. PMID- 10468774 TI - Is there a place for the insertion of a JJ stent as a temporizing procedure for symptomatic partial congenital vesico-ureteric junction obstruction in infancy? PMID- 10468775 TI - Incisional hernia after suprapubic catheter insertion. PMID- 10468776 TI - Renal calculus: a unique presentation of coeliac disease. PMID- 10468777 TI - Hazards of aspirin withdrawal before transurethral prostatectomy. PMID- 10468778 TI - Wegener's granulomatosis: prostatic involvement and recurrent urinary tract infections. PMID- 10468779 TI - Benign cystic mesothelioma of the tunica vaginalis. PMID- 10468780 TI - Dual drainage to salvage giant hydronephrosis: an innovative approach. PMID- 10468781 TI - Atypical Peyronie's disease. PMID- 10468782 TI - An unusual (knotty) complication of clean intermittent self-catheterization in a patient with an augmented bladder. PMID- 10468783 TI - Transient renal segmental ischaemia. PMID- 10468785 TI - Editor's choice PMID- 10468784 TI - Transobturator vascular graft biopsy at cystoscopy. PMID- 10468786 TI - Clinical measures of disease severity and outcome in psoriasis: a critical appraisal of their quality. AB - In clinical trials, a wide range of outcome measures has been used to evaluate the severity of psoriasis and its response to treatment. Despite their widespread use, many measures have received little attention with regards to their reliability and validity. Selecting an appropriately developed measurement tool is therefore of critical importance. We conducted a literature survey to examine the status of clinical outcome measures used in psoriasis research. The measures most commonly used were individual sign scores, e.g. for erythema, plaque thickness or scaling, and pooled indices, e.g. the Psoriasis Area and Severity Index. None of these, however, systematically fulfilled all the requirements of a validated instrument for disease assessment. Ideally, a core set of reliable and validated outcome measures for use in all psoriasis clinical trials is needed. Objective instrumental methods should minimise observer variation, but unless a simple non-invasive method can be developed, the uptake of such technology will probably be limited by cost and lack of practicality. Moreover, the translation of instrumental readings into clinically relevant measures is always a major problem, and for none of the methods has there been a robust mapping of instrumental readings on to a clinically meaningful scale. Further research is needed to determine the most appropriate and sensitive parameters to use as surrogate measures for capturing the distress which psoriatic patients feel but which is not measured with sufficient sensitivity or precision with current quality of life or distress questionnaires. PMID- 10468787 TI - Tumour necrosis factor-alpha induces Langerhans cell migration in humans. AB - The role of tumour necrosis factor (TNF)-alpha in the mobilization and migration of human epidermal Langerhans cells (LC) has been investigated. Intradermal injection of normal human volunteers with homologous recombinant TNF-alpha was found to cause a dose-dependent reduction in the frequency of LC within epidermal sheets 2 h later. Equivalent results were obtained when epidermal LC were identified on the basis of either CD1a or HLA-DR expression. At the dose of TNF alpha used routinely (500 U), treatment resulted in an average reduction in LC density of approximately 24%. Treatment with TNF-alpha was associated with a perivascular polymorphonuclear infiltration at 2 h, but the epidermis appeared normal with neither fibrinoid necrosis nor vasculitis, and LC morphology was not affected significantly. These results demonstrate that TNF-alpha provides an important signal for LC migration in humans and is likely therefore to play a crucial part in the induction of cutaneous immune responses. PMID- 10468788 TI - Fas/Fas ligand-mediated elimination of antigen-bearing Langerhans cells in draining lymph nodes. AB - Epidermal Langerhans cells (LC) are potent antigen-presenting cells (APC) that play a crucial part in initiating cutaneous immune responses. Although functional roles of LC as APC in the draining lymph node have been well investigated, little is known about the fate of LC after the antigen presentation to T cells. In this report, we demonstrate that antigen-bearing LC that migrated into the draining lymph nodes and were identified as fluorescent cells after skin painting with fluorescein isothiocyanate also expressed the Fas antigen. Clearance of the antigen-bearing LC was significantly delayed and the ratio of dead cells was reduced in Fas-deficient lpr and Fas ligand (FasL)-deficient gld mice at 2 days after skin painting, suggesting the involvement of a Fas/FasL-mediated pathway in the elimination of antigen-bearing LC in draining lymph nodes. These results suggest that a substantial population of LC after antigen presentation may undergo Fas/FasL-mediated apoptosis and that the elimination of active APC may be important for preventing excess immune responses. PMID- 10468789 TI - The repopulation of murine Langerhans cells after depletion by mild heat injury. AB - We have developed a model of focal Langerhans cell depletion by mild heat injury and used it to investigate the mechanisms of Langerhans cell repopulation in the injured epidermis. The possibility whether repopulation occurred by recruitment of precursor cells from the circulation or dermis or, alternatively, by migration from the surrounding normal epidermis into the injured area was considered. Repopulation was studied by evaluating the pattern of Langerhans cell reappearance and calculating the rate of change in the density. Heat injury followed by whole-body irradiation with shielding of the injured skin was used to assess repopulation in the absence of bone marrow precursors. Using tritiated thymidine autoradiography, we also investigated whether the newly arrived Langerhans cells (be they from circulating precursors or surrounding normal epidermis) actually divide. The results showed that heat injury completely eliminated the Langerhans cells within the area delineated by the injury. Two hours after injury, the Langerhans cells were fragmented and 2 days later, they could not be detected. Regeneration of the epidermis occurred 2 days after injury and Langerhans cells reappeared scattered somewhat sparsely in the centre of the lesion on day 3. These cells were small and slender, bearing one or two short dendrites. As the dendrites increased in number and in length, the cells became similar morphologically and phenotypically to normal Langerhans cells. The rate of repopulation increased dramatically between days 5 and 7 and reached normal density on day 11. The pattern of Langerhans cell repopulation in the injured area and the lack of repopulation in the irradiated animals indicated that repopulation occurs by immigration of precursors from the circulation or dermis. There was no indication of migration of Langerhans cells from surrounding normal epidermis. Lastly, the newly arrived Langerhans cells failed to divide at the site of injury. PMID- 10468790 TI - Differentiation of normal human keratinocytes influences hexavalent chromium uptake and distribution and the ability of cells to withstand Cr(VI) cytotoxicity. AB - The degree of differentiation of normal human keratinocytes determines the biology of the cells to a large extent. We have previously documented that keratinocytes from different donors differ significantly in their ability to withstand hexavalent chromium [Cr(VI)]-induced cytotoxicity. Several factors may contribute to this differing donor sensitivity to Cr(VI). The aims of this study were to investigate to what extent keratinocyte differentiation might influence Cr(VI) uptake and the ability of cells to withstand Cr(VI)-induced cytotoxicity. Keratinocytes from different donors were cultured under identical conditions and exposed to Cr(VI) (as potassium dichromate) at different points during their maturation process. The degree of differentiation of the cells was assessed using a quantitative assay for involucrin and related to the Cr(VI) cytotoxicity experienced by the cells. Chromium content was measured in whole cell, cytosolic and particulate fractions. While proliferative keratinocytes exposed to Cr(VI) showed a high degree of cytotoxicity to dichromate exposure, the more differentiated cells showed significantly less cytotoxicity but a higher uptake of the metal ion into the cells. The relative percentage of cytosolic chromium was high in the proliferative cells and decreased as the cells matured, suggesting that differentiated cultures were binding most of the chromium to the particulate fraction. Total chromium also increased during differentiation. The use of the channel-blocking agent 4, 4'-diisothiocyanate-2-2'-stilbenedisulphonic acid confirmed the spatial differences of chromium accumulation in the phenotypically different cultures, in that it prevented Cr(VI) entry into the proliferative cells and attenuated dichromate cytotoxicity in these cultures, but had no effect on the Cr(VI) uptake in differentiated cells, nor did it reduce its cytotoxicity. These data support the hypothesis that the upper differentiated layers of the epidermis are able to offer considerable physical protection to the lower proliferative layers from chemical pro-oxidants. PMID- 10468791 TI - Fibroblast expression of collagen integrin receptors alpha1beta1 and alpha2beta1 is not changed in systemic scleroderma. AB - The skin of patients with systemic scleroderma (SSc) is characterized by excessive extracellular matrix deposition in the dermis. As collagens represent the major structural component, we used fluorescence-activated cell sorter analysis to study the levels of collagen receptors expressed at the surface of fibroblasts derived from involved skin areas. In contrast to previous reports, no differences in the expression of alpha1, alpha2 or beta1 integrin subunits, which constitute the major collagen receptors on fibroblasts, were detected on SSc fibroblasts as compared with normal control fibroblasts. Variation of cell culture conditions, e. g. passage number (from 2 to 10), seeding density, cell cycle or serum concentration, did not change this result. These observations indicate that any abnormal response of SSc fibroblasts to their matrix environment is not controlled at the level of receptor expression. PMID- 10468792 TI - Internalization of gap junctions in benign familial pemphigus (Hailey-Hailey disease) and keratosis follicularis (Darier's disease). AB - Hereditary skin disorders involving acantholysis, such as Hailey-Hailey disease and Darier's disease, have been genetically linked to distinct chromosomal parts which do not code for known structural proteins. Such evidence suggests that the genomic abnormalities underlying these dermatoses may concern functional/regulatory mechanisms of keratinocyte cohesion. Epidermal communication junctions (gap junctions) are responsible for direct coupling of cells and, thus, co-ordinate the behaviour of keratinocytes within the tissue. Consequently, they remain one of the potential, and poorly studied, elements in the pathogenesis of hereditary acantholytic diseases. We have investigated the distribution and fate of gap junctions during non-immune acantholysis, using fine immunolocalization methods at the light and electron microscopic levels. Our results demonstrate normal expression of epidermal gap junction proteins, connexins 26 and 43, in non-lesional skin of Hailey-Hailey and Darier's diseases. The gap junctions were not primarily dismantled during acantholysis, typical of both of the studied dermatoses, but underwent internalization and subsequent cytoplasmic dispersion in the portions of cells which were no longer attached to the rest of the tissue. In Darier's disease, perifollicular acantholysis did not specifically concern epithelium of appendages coexpressing connexin 26 in addition to connexin 43, further indicating that the observed changes in gap junction localization were secondary to the loss of cell-cell contact. We demonstrated that the sequence of changes was identical in both diseases and that the previously described putative differences were apparently related to the degree of acantholysis present in the studied biopsies. The fate of the junctional structures and proteins, documented in the present study, is most probably a form of recycling process also used by normal keratinocytes during organogenesis and tissue differentiation. PMID- 10468793 TI - Forms of epithelial differentiation of draining sinus in acne inversa (hidradenitis suppurativa). AB - The draining sinus is a late complication of several forms of severe acne, leading to extensive, periodically inflamed lesions that are undermined by a system of fistulas, supposed to be of follicular origin. We investigated the expression of various cytokeratins (CKs) and desmosomal proteins in the draining sinus of acne inversa (hidradenitis suppurativa) using monoclonal antibodies in immunohistochemistry on paraffin-embedded sections. We were able to define three different phenotypes of stratified squamous epithelia covering the sinus tracts. Type I epithelium was cornifying and characterized by the presence of CK 10, desmogleins 1-3 and desmocollins 1-3 in an epidermis-like pattern. Type II epithelium was non-cornifying, negative for CK 10 and positive for CK 13. It was negative for desmocollin 1 but strongly immunopositive for desmoglein 1 suprabasally and for desmoglein 2 in the basal cells. Type III epithelium was non cornifying and strongly inflamed. It was marked by the presence of CK 7, CK 19 and desmoglein 2 and the absence of CK 10, desmoglein 1 and desmocollin 1. In both type II and III epithelium, desmoglein 3, desmocollin 2 and desmocollin 3 showed an inverted staining pattern as compared with normal epidermis and type I epithelium. Desmoglein 2 and CK 5/14 always remained restricted to the basal cell layer. Antibodies against CK 6 and CK 13/15/16 were immunopositive in all three phenotypes and CK 17 was predominantly immunolocalized to suprabasal layers of type II and III epithelium. The three phenotypes are characterized as pathological stratified squamous epithelia reflecting the dynamic process of inflammation, proliferation and stratification taking place in acne inversa. PMID- 10468794 TI - Development of an oligonucleotide probe specific for Trichophyton rubrum. AB - A species-specific DNA probe was developed to detect the dermatophyte species Trichophyton rubrum. The selected oligonucleotide sequence is derived from the highly variable internal transcribed spacer 2 region of the ribosomal DNA operon. The specificity of the non-radioactive labelled oligonucleotide probe was tested against related dermatophytes, other eukaryotic microorganisms and against human DNA. No cross-hybridization was found, and hybridization signals were invariably detected in all T. rubrum strains investigated. In addition, no homologous sequences were found searching the EMBL database. Experiments to establish a method for isolating DNA directly from clinical specimens gave successful amplification and hybridization products in about 30% of the samples. PMID- 10468795 TI - Human papillomavirus type 5 is commonly present in immunosuppressed and immunocompetent individuals. AB - DNA of a wide spectrum of epidermodysplasia verruciformis-associated human papillomavirus (HPV) types (EV-HPV) has been detected in skin lesions and plucked hairs from both immunosuppressed and a considerable proportion of non immunosuppressed persons. Recently, the skin of psoriatic patients was claimed to be an important reservoir for a particular EV-HPV type, HPV 5, which is considered as a high-risk HPV type for skin carcinomas. In the present study, we analysed plucked hairs from immunosuppressed renal transplant patients and immunocompetent individuals, utilizing an HPV 5-specific nested polymerase chain reaction. HPV 5 was detected in hairs derived from 14 of 31 (45%) immunosuppressed patients and 21 of 135 (16%) immunocompetent individuals. Both the immunosuppressed and the immunocompetent groups consisted of individuals with and without non-melanoma skin cancer. HPV 5 DNA was detected in similar proportions of hair samples plucked from individuals with and without skin cancer in either group. Our results indicate that HPV 5 is commonly present in the population. The role of HPV 5 in the pathogenesis of skin carcinomas and psoriasis remains to be established. PMID- 10468796 TI - A mineral sunscreen affords genomic protection against ultraviolet (UV) B and UVA radiation: in vitro and in situ assays. AB - Ultraviolet (UV) radiation has been shown to be responsible for different biological effects on human skin, including the initiation of photocarcinogenesis. Both UVB and UVA have been described as mutagenic, but the processes by which they alter the DNA are different. Although cells can repair DNA damage, some deleterious mutations nevertheless appear and can promote cancer. The risk of photocarcinogenesis is acknowledged and the frequency of photogenodermatosis is increasing. In order to evaluate the protection efficacy of a high sun protection factor (SPF) mineral sunscreen against UVB- and UVA induced genomic alterations, we have followed two approaches. First, we have tested the sunscreen for its ability to decrease the unscheduled DNA synthesis response in vitro in human fibroblasts, as an indirect measure of UVB-induced lesions (0.005 and 0.01 J/cm2), and second, we have verified its ability to reduce the in situ end-labelling intensity in human skin as a direct measure of UVA-induced single-strand breaks (10 J/cm2). Microscopic analysis clearly demonstrated the protective effect of the sunscreen against UVB and UVA. A dose dependent effect of mineral sunscreens was observed. There was also a relationship between the SPF and genomic protection. By limiting the accumulation of UV-induced lesions on DNA, this mineral sunscreen could limit the mutation frequency. PMID- 10468797 TI - A prospective coagulation study including resistance to activated protein C and mutations in factors V and II in venous leg ulcers. AB - Hypercoagulable states have been reported to be associated with venous leg ulcers. In an attempt to investigate the prevalence of hypercoagulable states in patients with venous leg ulcers, we performed a prospective case-control study for the presence of coagulation defects in such patients, including resistance to activated protein C (APC), factor V Leiden mutation and a newly described mutation in factor II. Results were compared with those obtained in a control group. APC resistance was found in four of 33 patients tested, but only one was found to be heterozygous for the factor V Leiden mutation. Factor II mutation was found in two of 30 patients tested. Our findings show that the prevalence of coagulation abnormalities is not different in patients with venous leg ulcers from controls in our study, suggesting that only selected patients with venous ulcers and a history of recurrent deep venous thrombosis should be investigated for the presence of coagulation defects. PMID- 10468798 TI - A novel anti-inflammatory drug, SDZ ASM 981, for the treatment of skin diseases: in vitro pharmacology. AB - SDZ ASM 981, a novel ascomycin macrolactam derivative, has high anti-inflammatory activity in animal models of allergic contact dermatitis and shows clinical efficacy in atopic dermatitis, allergic contact dermatitis and psoriasis, after topical application. Here we report on the in vitro activities of this promising new drug. SDZ ASM 981 inhibits the proliferation of human T cells after antigen specific or non-specific stimulation. It downregulates the production of Th1 [interleukin (IL)-2, interferon-gamma] and Th2 (IL-4, IL-10) type cytokines after antigen-specific stimulation of a human T-helper cell clone isolated from the skin of an atopic dermatitis patient. SDZ ASM 981 inhibits the phorbol myristate acetate/phytohaemagglutinin-stimulated transcription of a reporter gene coupled to the human IL-2 promoter in the human T-cell line Jurkat and the IgE/antigen mediated transcription of a reporter gene coupled to the human tumour necrosis factor (TNF)-alpha promoter in the murine mast-cell line CPII. It does not, however, affect the human TNF-alpha promoter controlled transcription of a reporter gene in a murine dendritic cell line (DC18 RGA) after stimulation via the FcgammaRIII receptor. SDZ ASM 981 also prevents the release of preformed pro inflammatory mediators from mast cells, as shown in the murine cell line CPII after stimulation with IgE/antigen. In summary, these results demonstrate that SDZ ASM 981 is a specific inhibitor of the production of pro-inflammatory cytokines from T cells and mast cells in vitro. PMID- 10468799 TI - Topical maxacalcitol for the treatment of psoriasis vulgaris: a placebo controlled, double-blind, dose-finding study with active comparator. AB - 1alpha, 25-Dihydroxy-22-oxacalcitriol (maxacalcitol) is a vitamin D3 analogue which displays approximately 10 times greater efficacy at suppressing keratinocyte proliferation in vitro than calcipotriol and tacalcitol. To determine clinical efficacy, a phase II double-blind, randomized, left vs. right, concentration-response study was performed with once-daily topical maxacalcitol in patients with mild to moderate chronic plaque psoriasis. Primary efficacy parameters were psoriasis severity index (PSI) based on sum of scores for erythema, scaling and induration and investigators' overall assessment of patients' response to therapy at 8 weeks of treatment. One hundred and forty-four patients participated. All concentrations of maxacalcitol ointment (6, 12.5, 25 and 50 microg/g) were significantly more effective at reducing PSI than placebo (P < 0.01), with greatest effect noted for maxacalcitol 25 microg/g. Calcipotriol ointment 50 microg/g once daily as active comparator had a similar effect. Marked improvement or clearance of psoriasis was greatest for maxacalcitol 25 microg/g (55% of subjects) which compared favourably with calcipotriol (46%). Improvement continued throughout the study period, with no plateau at week 8. Investigators' and patients' side preference (secondary efficacy parameters) rated maxacalcitol more effective than placebo and 25 microg/g maxacalcitol better than calcipotriol (P < 0.05 for investigators' assessment). Twelve patients withdrew from the study due to adverse events, of which four were judged to be due to study medication. This study indicates that once-daily maxacalcitol ointment is effective in the management of plaque psoriasis, with greatest effect noted at 25 microg/g. As no response plateau was noted at 8 weeks, these data suggest that further benefit might be obtained if maxacalcitol ointment were applied for longer. Finally, investigators' overall assessment and side preference suggest that maxacalcitol 25 microg/g may be more effective than once-daily calcipotriol. PMID- 10468800 TI - Combination treatment with methotrexate and cyclosporin for severe recalcitrant psoriasis. AB - An increasingly important approach to the management of patients with severe psoriasis is the concurrent use of two systemic treatments. Previous guidelines have advised against the use of methotrexate and cyclosporin in combination. We report the successful use of a combination of methotrexate and cyclosporin in the treatment of 19 patients with severe, recalcitrant psoriasis, 15 of whom had psoriatic arthropathy. Most patients had previously received two or more systemic treatments. Before combination treatment was started nine of the patients were taking methotrexate and 10 were taking cyclosporin at the maximum tolerated doses. The duration of combination treatment was bimodally distributed, with seven patients having short-term treatment (mean +/- SD duration 18. 9 +/- 15.7 weeks) and 12 patients having long-term treatment (mean +/- SD duration 193.2 +/- 160.6 weeks). Those patients who received short-term treatment did not develop any evidence of toxicity from either agent. Of those patients on long-term treatment, three developed mild impairment of renal function that returned to normal following a reduction in dose of cyclosporin, and three had impairment of renal function (following long-term cyclosporin monotherapy) that improved, but did not normalize, following a reduction in dose of cyclosporin. In each case, combination treatment for psoriasis resulted in good control of both skin and joint problems using lower doses of each agent than would have been used for monotherapy. We conclude that the combination of methotrexate and cyclosporin is an effective treatment for this group of patients. PMID- 10468801 TI - Intermittent short courses of cyclosporin (Neoral(R)) for psoriasis unresponsive to topical therapy: a 1-year multicentre, randomized study. The PISCES Study Group. AB - We performed a 1-year study to determine whether intermittent short courses of the microemulsion formulation of cyclosporin (Neoral) could effectively control plaque psoriasis and whether tapering or abrupt cessation of cyclosporin therapy would influence time to relapse. Four hundred patients with plaque psoriasis were included in this open, multicentre, randomized study. All patients commenced cyclosporin at a dose of 2.5 mg/kg daily. Cyclosporin dosage could be increased to a maximum of 5 mg/kg daily. Treatment was continued until clearance of psoriasis or for a maximum of 12 weeks. Patients were then randomly assigned either to stop cyclosporin abruptly or to have the dose reduced by 1 mg/kg daily each week until cessation. On relapse, patients were given another course of cyclosporin. Patients were followed for at least 1 year, during which they could receive as many treatment courses as necessary. The number of patients who required one, two, three and four treatment courses was 400, 259, 117 and 26, respectively. The median time to relapse after the end of the first treatment period was 109 days in the group of patients randomized to stop cyclosporin abruptly and 113 days in patients randomized to taper off cyclosporin (P = 0.038). More than 30% of patients had not relapsed 6 months after having stopped treatment. After each treatment course, the Kaplan-Meier probability of achieving 75% or more reduction in disease area by day 84 of treatment was 83%, 76%, 73% and 66%, respectively. Mean serum creatinine concentration and blood pressure did not show any clinically significant changes over time. Our results show that intermittent short-course therapy with Neoral, when used in conjunction with topical therapy, is well tolerated and provides effective control of plaque psoriasis for 1 year. Tapering off cyclosporin on treatment cessation induces a slight delay in psoriasis relapse. PMID- 10468802 TI - Recalcitrant viral warts treated by diphencyprone immunotherapy. AB - Recalcitrant viral warts are a troublesome therapeutic problem. Immunotherapy with the universal allergic contact sensitizer diphencyprone (DCP) has been used successfully in such cases. We have reviewed our experience of the use of DCP in the treatment of resistant hand and foot warts during an 8-year period. Sixty patients were sensitized to DCP during this time; the median duration of warts was 3 years. Twelve patients defaulted from treatment. Of the remaining 48 individuals, 42 (88%) cleared of all warts. The median number of treatments to clear was five (range one to 22) and the median time to clear was 5 months (range 0.5-14). Adverse effects occurred in 27 of 48 patients (56%), most commonly painful local blistering (n = 11), blistering at the sensitization site (n = 9), pompholyx-like reactions (n = 7) and eczematous eruptions (n = 4). Three of those who defaulted did so due to side-effects, one became pregnant and eight dropped out for unknown reasons. Three of the 48 patients who cleared or had at least six treatments also discontinued DCP therapy due to side-effects, but most tolerated treatment well. Twenty-five patients were followed up for periods of 1 month to 8 years (median 2 years) and none had a recurrence. DCP immunotherapy is an effective option for the treatment of recalcitrant viral warts but patients must be motivated to attend for sequential applications and must be warned about potential uncomfortable side-effects. PMID- 10468803 TI - The familial risk of adult acne: a comparison between first-degree relatives of affected and unaffected individuals. AB - Acne is principally a disorder of adolescence but persists into middle age in a small minority of individuals. There is some evidence, primarily from twin studies, to suggest that acne may be an inherited disease. We have carried out an investigation of the familial risk of persistent adult acne by studying the occurrence of this condition in first-degree relatives of patients with adult persistent acne compared with the relatives of unaffected matched controls. Two hundred and four patients over the age of 25, referred to our department with facial acne which had persisted from adolescence, were included in the study. For each patient, a detailed pedigree which included all first-degree relatives was drawn up. For all relatives over the age of 25, demographic details and the presence or absence of facial acne was recorded. One hundred and forty-four unaffected controls were recruited. These were matched with the patient group in terms of age, social class and ethnic origin. The same questions were asked of controls as of the cases, and they provided the same information on their first degree relatives. In total, 204 acne cases and 144 non-acne control volunteers were studied, contributing 1203 and 856 first-degree relatives, respectively. Two hundred and three first-degree relatives of cases were affected with facial acne, compared with 42 first-degree relatives of volunteers. The risk of adult acne occurring in a relative of a patient with adult acne was significantly greater than for the relative of an unaffected individual (odds ratio 3.93, 95% confidence interval 2.79-5.51; P < 0.001). Our study suggests that familial factors are important in determining individual susceptibility to adult persistent facial acne. Genetic factors may determine the failure of acne-prone follicles to evolve into acne-resistant follicles in early adult life. PMID- 10468804 TI - Serum selenium levels and blood glutathione peroxidase activities in vitiligo. AB - It has recently been shown that patients with vitiligo can accumulate epidermal hydrogen peroxide (H2O2) in association with low catalase levels. This study examined serum selenium levels and blood glutathione peroxidase activities in 61 patients and controls. The results showed high serum selenium levels in 56% of the patients. As at least one isoform of glutathione peroxidase requires selenium for its activity, enzyme activities were also evaluated. The overall results were not significantly different compared with controls, but further age-related analysis of the data indicated significantly lower activities in patients up to 46 years. As glutathione peroxidase can also efficiently degrade H2O2, the results of this study could indicate an additional impaired H2O2 metabolism in vitiligo. PMID- 10468806 TI - The treatment of acne fulminans: a review of 25 cases. AB - The treatment of acne fulminans has been difficult. It is difficult to perform a controlled treatment trial due to the rarity of the complication. However, it is possible to compare four different therapeutic regimens which have evolved with time in the management of 25 patients over a period of 25 years. Oral antibiotics produced a slow response in the resolution of acne and systemic symptoms. The addition of a systemic steroid produced a quick resolution of systemic features, but the time until resolution of the acne was longer than when it was used in combination with oral isotretinoin. The protocols which used a combination of prednisolone and isotretinoin led to faster control of systemic features as well as clearance of acne when compared with other protocols. This was particularly so if the oral steroid was used sooner rather than later. We conclude that the preferred treatment of acne fulminans is oral prednisolone 0.5-1 mg/kg daily for 4-6 weeks (thereafter slowly reduced to zero) with oral isotretinoin being added to the regimen at the fourth week, initially at 0.5 mg/kg daily and gradually increased to achieve complete clearance. PMID- 10468805 TI - Intermittent short duration therapy with fluconazole is effective for tinea capitis. AB - We have demonstrated in an open multicentre investigation that oral fluconazole 6 mg/kg daily for 2 weeks, followed, if clinically indicated four weeks from the start of therapy, by an extra week of treatment at the same dosage, may be effective and safe in the treatment of tinea capitis. Of a total of 48 patients, there were 42 evaluable children < 18 years old (19 boys, 23 girls; mean age 6.2 years, range 1.5-16). The causative organisms were Trichophyton tonsurans (38 subjects) and T. violaceum (four). In the 42 evaluable patients, a 2-week course of fluconazole was administered in 21, with the remainder requiring 1 additional week of therapy. At follow-up 12 weeks from the start of therapy, mycological and clinical cure was recorded in 37 of the 42 evaluable patients (88.1%, 95% confidence interval 83.1-93.1%). The treatment was well tolerated, with no clinical adverse effects. This regimen appears to be effective and safe, and is associated with high compliance. The preliminary results of the investigation need to be evaluated in a larger sample of patients, and in tinea capitis caused by zoophilic species. PMID- 10468807 TI - Transepidermal elimination of urate-like crystals: a new perforating disorder? AB - We report two men who developed a transient perforating disorder characterized by transepidermal elimination of negatively birefringent needle-shaped crystals similar to monosodium urate. This striking clinical presentation has not previously been described and we propose that it be added to the group of diseases known as the primary perforating disorders. PMID- 10468808 TI - Follicular cutaneous T-cell lymphoma: a clinicopathological study of nine cases. AB - Nine patients with follicular cutaneous T-cell lymphoma (CTCL), a recently described variant of lymphoma, are presented. On the basis of clinical manifestations and disease course, three groups of patients were distinguished: (i) two patients with follicular CTCL not associated with conventional lesions of mycosis fungoides (MF) and showing no evolution towards MF in follow-up periods of 3 and 6 years; (ii) one patient with follicular CTCL that evolved into conventional MF within 3 years; (iii) six patients showing conventional MF lesions either before or concurrently with the follicular lesions and thus representing follicular CTCL of the true MF type. The follicular lesions included hair-devoid patches or plaques with spiky hyperkeratotic papules (four patients), keratosis pilaris-like lesions (four), comedo-like lesions (four), follicular papules with alopecia (three) and milia-like lesions (three). Histopathological examination showed perifollicular and intrafollicular lymphocytes, without mucin deposition and with minimal or no involvement of the overlying epidermis. Significant syringotropism was also observed in three cases. Immunohistochemical analysis showed the predominance of CD4 + T cells, deletion of CD7 in some cases, Ki-67 + lymphocytes confined mainly to the follicular epithelium, and expression of keratinocyte intercellular adhesion molecule-1 exclusively in the hair follicle. T-cell receptor gamma gene rearrangement was positive in the one case studied from each group. Different treatment modalities were employed, the most commonly used as monotherapy being phototherapy: psoralen ultraviolet A in four patients, two of whom showed a complete clinical and histopathological remission, and ultraviolet B in one patient, who showed a complete remission (both clinical and histopathological). This study indicates that follicular CTCL is more common than reflected in the literature, has heterogeneous clinical manifestations, and is either an expression of or closely related to MF. The influence of the follicular involvement on the therapeutic response remains to be clarified. However, our therapeutic experience clearly suggests that some patients with follicular CTCL can benefit from phototherapy. PMID- 10468809 TI - Microbiological and molecular diagnosis of deep localized cutaneous infection with Trichophyton mentagrophytes. AB - We describe a healthy young woman with a localized deep dermal infection on the right side of the chest wall. It was caused by the dermatophyte Trichophyton mentagrophytes, and resolved after two pulses of oral itraconazole 200 mg twice daily for 1 week. As cultural and microscopic features did not enable a precise identification of the fungus, molecular investigation was undertaken. Patterns of HaeIII restriction digests of genomic DNA from the culture matched those from Arthroderma incurvata and A. benhamiae, which is the teleomorph of T. mentagrophytes var. mentagrophytes. PMID- 10468810 TI - Lipoid proteinosis. A biochemical and ultrastructural investigation of two new cases. AB - Lipoid proteinosis is a rare autosomal recessive disease characterized by cutaneous and visceral lesions, in which large amounts of amorphous material are constantly found in stroma. Morphological and biochemical studies indicate abnormal collagen production, but little attention has been paid to the lipid component of lesions. Microscopic and ultrastructural studies of skin, with special emphasis on fibroblasts, vessels, nerve endings and eccrine sweat glands, were conducted in two patients with lipoid proteinosis. Biochemical studies were undertaken in cultured fibroblasts. Evidence of lysosomal storage in epithelial cells of eccrine sweat glands and in dermal histiocytes, very similar to that found in some metabolic disorders, particularly Farber disease, was found in both cases. Our findings suggest that two alterations might coexist in lipoid proteinosis, one characterized by impaired normal collagen production and the other related to a metabolic defect which may lead to accumulation of ceramide or more complex lipids. PMID- 10468811 TI - Congenital annular multiple fibrofolliculomas occurring with deformity of the ear and ventricular septal defect. AB - We describe a 5-year-old girl who had multiple fibrofolliculomas with an unusual annular configuration, present since birth, localized to the mid-back. She had no family history of similar skin lesions. Examination showed a depigmented patch on her left buttock and other congenital anomalies, i.e. deformity of the auricle of the ear and ventricular septal defect. There has been no previous report of congenital multiple fibrofolliculomas occurring with congenital malformations such as deformity of the auricle of the ear and ventricular septal defect. The congenital occurrence and unusual configuration of the lesions in our patient may suggest a naevoid origin for these tumours. PMID- 10468812 TI - Erythema elevatum diutinum and HIV infection: a report of five cases. AB - Erythema elevatum diutinum (EED) is emerging as a specific HIV-associated dermatosis, 11 cases having so far been reported in the medical literature and five patients with the disease having been seen by us during the last 4 years. As the disease is poorly known, it is easily confused with Kaposi's sarcoma or bacillary angiomatosis, but the histopathological features are diagnostic. EED is considered to be an immune complex-mediated vasculitis. A streptococcal infection seemed to be the trigger factor in four of our patients. Partial control of the cutaneous lesions was achieved by the use of antibiotics. PMID- 10468813 TI - Pyoderma gangrenosum associated with Takayasu's arteritis responding to cyclosporin. AB - A 33-year-old caucasian woman with pyoderma gangrenosum associated with Takayasu's arteritis responded to treatment with cyclosporin. This patient is unusual in that both ulcerative and vesiculopustular forms of pyoderma gangrenosum were present. This has not previously been reported with Takayasu's arteritis. PMID- 10468814 TI - The peripheral corneal melting syndrome and psoriasis: coincidence or association? AB - The peripheral corneal melting syndrome (PCMS) is a rare disease consisting of marginal corneal thinning that can progress to perforation. The PCMS carries a grave prognosis and it is of vital importance that dermatologists are aware that this may be responsible for a painful red eye in a patient with psoriasis. We highlight the features of the PCMS developing in an elderly woman with long standing psoriasis to increase awareness of its significance, and hypothesize that an association may exist between the two conditions. Only one previous report has been published in which the authors speculate on the possible association of this syndrome with psoriasis. That few other cases have been described is either a consequence of under-reporting by both ophthalmologists and dermatologists unaware of a link or because the relationship between the syndrome and psoriasis is genuinely coincidental. PMID- 10468815 TI - Photodynamic therapy for inoperable vulval Paget's disease using delta aminolaevulinic acid: successful management of a large skin lesion. AB - A 74-year-old woman with extensive inoperable vulval extramammary Paget's disease who had lymph node and pulmonary metastases, was treated with etoposide (VP16) 100 mg and 5000 cGy electron beam irradiation, which reduced the lesion by 60%. Following these interventions, the residual lesion was successfully treated with repeated photodynamic therapy using delta-aminolaevulinic acid, which achieved a nearly complete remission. PMID- 10468816 TI - Classification of primary cutaneous B-cell lymphomas: EORTC classification or REAL classification? PMID- 10468817 TI - Primary cutaneous B-cell lymphoma: how useful is the new European Organisation for Research and Treatment of Cancer Classification? PMID- 10468819 TI - Classification of primary cutaneous B-cell lymphomas PMID- 10468818 TI - Classification of primary cutaneous B-cell lymphomas. PMID- 10468820 TI - The application of microwave irradiation in immunofluorescence for the diagnosis of autoimmune bullous disease. PMID- 10468821 TI - Paraneoplastic pemphigus with cutaneous and serological features of pemphigus foliaceus. PMID- 10468822 TI - T-cell epitope mapping performed after patch testing in a patient with contact allergy to several azo dyes. PMID- 10468823 TI - Failure to detect human herpesvirus type 8-DNA sequences in systemic plasmacytosis from HIV-uninfected Japanese patients. PMID- 10468824 TI - Immunohistochemistry and sentinel node biopsy in melanoma: the use of tyrosinase and Melan-A. PMID- 10468825 TI - Cutaneous manifestations of Sjogren's syndrome associated with myasthenia gravis. PMID- 10468826 TI - Chondrodermatitis nodularis helicis in childhood dermatomyositis. PMID- 10468827 TI - Dermatomyositis sine myositis presenting with calcinosis universalis. PMID- 10468828 TI - Seasonally recurrent granuloma annulare on sun-exposed areas. PMID- 10468829 TI - Halo naevus with histological changes resembling epidermal erythema multiforme. PMID- 10468830 TI - Amelanotic acral lentiginous malignant melanoma. PMID- 10468831 TI - Leukaemia cutis and erythroleukaemia. PMID- 10468832 TI - Cutaneous metastasis from an adenoid cystic carcinoma of the lacrimal gland. PMID- 10468833 TI - Solitary giant vulval syringoma. PMID- 10468834 TI - Bullous keloid: a distinct entity? PMID- 10468835 TI - Familial naevus sebaceus may be explained by paradominant transmission. PMID- 10468836 TI - Primary cutaneous aspergillosis. PMID- 10468837 TI - Kaposi-like acroangiodermatitis in an amputee. PMID- 10468838 TI - Oral presentation of a novel variant of herpes simplex infection in a group of bone marrow transplant patients: a report of five cases. PMID- 10468839 TI - Basal cell carcinomas occurring after accidental exposure to ionizing radiation. PMID- 10468840 TI - Photodynamic therapy with meso-tetra(hydroxyphenyl) chlorin in the topical treatment of Bowen's disease and basal cell carcinoma. PMID- 10468841 TI - Seizures associated with interferon-alpha treatment of cutaneous malignancies. PMID- 10468842 TI - Successful immunotherapy of chronic nodular prurigo with topical dinitrochlorobenzene. PMID- 10468843 TI - Linear porokeratosis of mibelli: successful treatment with cryotherapy. PMID- 10468844 TI - Somatotrophin-induced acanthosis nigricans. PMID- 10468845 TI - Severe toxic contact dermatitis caused by garlic. PMID- 10468846 TI - Malaria in an African penguin. PMID- 10468847 TI - Detection of minimal residual disease in acute myeloid leukaemia: methodologies, clinical and biological significance. PMID- 10468848 TI - The pathogenesis of acute promyelocytic leukaemia: evaluation of the role of molecular diagnosis and monitoring in the management of the disease. PMID- 10468849 TI - Mll rearrangements in haematological malignancies: lessons from clinical and biological studies. PMID- 10468850 TI - High levels of lung resistance related protein mRNA in leukaemic cells from patients with acute myelogenous leukaemia are associated with inferior response to chemotherapy and prior treatment with mitoxantrone. AB - Expression of the mdr1 (multidrug resistance), mrp (multidrug resistance associated protein), and lrp (lung resistance related protein) genes is associated with transport related MDR (multidrug resistance). We quantified mRNA levels of these genes using competitive reverse transcription polymerase chain reaction (RT-PCR) in 128 samples of leukaemic cells from 92 patients with acute myelogenous leukaemia (AML). There was a wide variation between the samples in mRNA levels of all three genes. The mean mdr1 mRNA level was 1.3 transcripts per cell (range undetectable to 15.8), the mean mrp level was 7.9 (range 0.1-36.2) and mean lrp 3.9 (range 0.1-29). Lrp mRNA levels were higher in samples drawn at diagnosis from the 15 patients with resistant disease than from the 37 with chemosensitive disease (4.9 SD 3.1 v 2.9 SD 2.3, P = 0.016). Neither mdr1 nor mrp mRNA levels were predictive for response to chemotherapy. In samples from patients who had received chemotherapy, those that had received mitoxantrone (n = 24) had higher lrp mRNA levels (mean 4.8, SD 2.5) than those that had not (n = 20, mean 2.8, SD 2.4, P = 0.012). In conclusion, the results indicate that lrp expression is associated with inferior response to chemotherapy in AML and that lrp expression increases after exposure to mitoxantrone. PMID- 10468851 TI - Quantification of minimal residual disease in patients with BCR-ABL-positive acute lymphoblastic leukaemia using quantitative competitive polymerase chain reaction. AB - We analysed 20 patients with BCR-ABL-positive acute lymphoblastic leukaemia (ALL) by quantitative competitive polymerase chain reaction (QC-PCR) to study the kinetics of the leukaemic clone. Consecutive samples of 16 patients (minor-bcr, n = 10; major-bcr, n = 6) were analysed after conventional chemotherapy and/or bone marrow transplantation (BMT). DNA competitor templates co-amplifying with either p210 or p190 BCR-ABL cDNA were used for quantification of leukaemia-specific BCR ABL mRNA. In all samples, total ABL transcripts were measured as internal control, and the percentage of BCR-ABL/ABL molecules was calculated. Following induction chemotherapy the number of BCR-ABL transcripts was reduced by a maximum of 2-3 logs. In most patients, additional chemotherapy did not lead to further reduction of BCR-ABL mRNA. In two patients, conventional chemotherapy plus autologous BMT in complete haematological remission resulted in a total reduction of the transcript level of more than 3 logs. In two other patients, allogeneic BMT caused a transient reduction of the BCR-ABL transcripts below the detection level of our method (<1 blast cell in 105 normal cells) for a period of 7 and 11 months, respectively. The achievement of PCR negativity did not guarantee sustained remission. Both patients relapsed and BCR-ABL transcript levels rose by more than 1 log prior to frank relapse. Our data demonstrate that quantification of BCR-ABL mRNA allows the evaluation of the dynamics of the leukaemic clone and thus is valuable for the evaluation of minimal residual leukaemia following various therapies and the early detection of increasing BCR-ABL transcripts prior to relapse. PMID- 10468852 TI - The cyclin-dependent kinase inhibitors p18INK4c and p19INK4d are highly expressed in CD34+ progenitor and acute myeloid leukaemic cells but not in normal differentiated myeloid cells. AB - Cyclin-dependent kinase inhibitors (CKIs) are important for the differentiation of cells in various tissues. In acute myeloid leukaemia (AML) the cells accumulate at particular stages of myeloid maturation. We therefore analysed the expression pattern of different CKIs in fresh samples of AML patients and compared it with that in CD34+ progenitor and normal differentiated myeloid cells. Competitive RT-PCR and Western analysis revealed a significantly higher expression of p18INK4c and p19INK4d in leukaemic and CD34+ progenitor cells than in granulocytes and monocytes. A different pattern was seen for p27Kip1 and p57Kip2 expression being low in leukaemic cells but high in normal immature and differentiated cells. No marked differences were found in p15INK4b and p21Cip1 mRNA expression between leukaemic and CD34+ progenitor or mature myeloid cells. Our findings therefore indicate that high expression of p18INK4c and p19INK4d in haemopoietic progenitor and leukaemic blast cells may contribute to the premature differentiation block seen in AML. PMID- 10468853 TI - FADD expression and caspase activation in B-cell lymphomas resistant to Fas mediated apoptosis. AB - We have previously shown that malignant B cells from non-Hodgkin's lymphomas (NHL) are resistant to Fas-mediated apoptosis. To determine the mechanisms underlying this resistance, we analysed by Western blotting the expression of several apoptotic regulators, caspase 3, caspase 8, FADD and poly(ADP-ribose) polymerase (PARP) in fresh lymphoma cells, isolated from 16 B-NHL biopsy samples of different histological subtypes, and displaying variable levels of Fas expression. The profiles of expression of these apoptotic regulators were monitored in cell lysates at different times following Fas with or without CD40 stimulation. Expression of FADD and of the uncleaved forms of PARP, caspase 3 and caspase 8 were detected in all untreated NHL samples. Low levels of PARP cleavage were noted in three untreated samples. Fas stimulation alone induced neither significant apoptosis nor significant changes in the expression profiles of FADD, caspases 3 and 8 and PARP in the 16 samples, except for variations in FADD and caspase 8 expression levels in a minority of samples. Fas/CD40 co-stimulation induced apoptosis and cleavage of caspase 3, caspase 8 and PARP in the five NHLs tested; expression of FADD was not modified. Our results showed (1) that induction of apoptosis in B-NHLs by Fas/CD40 co-stimulation used the same caspase executioner machinery as the normal Fas pathway, and (2) that NHL cells which resisted Fas-mediated apoptosis displayed no defect in either expression or functionality of caspases 3 and 8, nor in FADD expression. The dysfunction underlying NHL resistance to apoptosis must therefore lie upstream of caspase 8, or could alternatively be influenced by anti-apoptotic regulators of the Bcl-2 family. PMID- 10468854 TI - Telomerase activity in chronic lymphoproliferative disorders of B-cell lineage. AB - The progressive shortening of telomeres at each cell division is a key mechanism in controlling cell proliferative capacity. The activation of telomerase, a reverse transcriptase that extends telomere length, potentially leads to unlimited cell proliferation, and is believed to play a critical role in the neoplastic process. High levels of telomerase activity have been demonstrated in almost all solid tumours; however, little data is available concerning its expression in chronic B-cell neoplasms. By using a quantitative polymerase chain reaction-based method we quantified telomerase activity in normal B lymphocytes, and in various B-cell malignancies, including chronic lymphocytic leukaemia (CLL), mantle cell lymphoma (MCL) and hairy cell leukaemia (HCL). Compared to normal B cells, which expressed very low levels of telomerase activity, malignant cells from most of the patients showed a significant increase in telomerase activity, with highest values observed in HCL samples. Moreover, among the CLL and HCL cases, significantly higher levels of telomerase activity were found in patients with progressive disease at 1 year follow-up versus patients with stable disease. These data suggest that telomerase activity might correlate with disease progression. PMID- 10468855 TI - Phenotypic and molecular analysis of six human cell lines derived from patients with plasma cell dyscrasia. AB - Cell lines RPMI 8226, JJN3, U266 B1, NCI-H929 (all EBV-) and ARH77 and HS-Sultan (both EBV+) have been extensively characterized in this study. EBV- lines expressed the phenotype (CD138-, CD19+, CD20+) whereas EBV+ were (CD138+, CD19-, CD20-). CD56 expression was restricted to EBV- cell lines, with the exception of U266 B1, whereas PCA-1 was strongly expressed on five of the six cell lines. Only EBV+ cell lines bound peanut-agglutinin (PNA). However, all cell lines bound the lectin Jacalin that binds the same receptor as PNA, irrespective of the receptors sialylation status. By RT-PCR and direct sequencing of their IgH V/D/J domains, ARH77 was demonstrated to use the germline sequence VH4-34/dm1/JH6b, whereas no arrangement was demonstrated for RPMI 8226, suggesting IgH gene deletion or mutation. HLA class I and II antigens were detected using HLA typing on all cell lines warranting their use as suitable targets for HLA-restricted cytotoxic T cells. By sensitive RT-PCR, mRNA for IL-6, IL-6R and TNFbeta was found expressed in all cell lines. IL-1 mRNA expression was predominantly associated with the EBV+ phenotype. Although mRNA for IL-3 and GM-CSF was never detected, transcripts for c-kit ligand and, more commonly, its receptor were. Likewise GM-CSF, M-CSF and erythropoietin mRNA transcripts were detected in the majority of cell lines. PMID- 10468856 TI - Bone marrow effects of anagrelide therapy in patients with myelofibrosis with myeloid metaplasia. AB - In a prospective study investigating the therapeutic role of anagrelide in myelofibrosis with myeloid metaplasia, 20 patients received anagrelide in daily oral doses of 0.5-3 mg. 17 patients were evaluable and received anagrelide for a median of 2 years (range 0.5-4 years). No patient had a clinically appreciable benefit. Bone marrow (BM) examinations at baseline and after 6 and 12 months of treatment were available for 17, 17 and 12 patients, respectively. In all evaluable cases, BM megakaryocyte number increased after 6 months of anagrelide treatment. Also, Ulex europaeus agglutinin-1 staining of megakaryocytes revealed a left-shifted maturation pattern in most patients with a platelet response to anagrelide. However, megakaryocyte staining intensity for transforming (TGF-beta) and platelet-derived (PDGF) growth factors was not affected consistently by treatment. No patient had a >/=2 grade change in either BM fibrosis or osteosclerosis. These in-vivo data support our previous in-vitro observations that anagrelide interferes with megakaryocyte maturation rather than proliferation. Lack of a positive treatment effect is consistent with the finding that anagrelide did not significantly alter megakaryocyte expression of TGF-beta and PDGF. PMID- 10468857 TI - Leucocyte transfusions from rhG-CSF or prednisolone stimulated donors for treatment of severe infections in immunocompromised neutropenic patients. AB - Sepsis in profound neutropenia after chemotherapy is associated with high mortality despite appropriate antibacterial or antifungal treatment. In a prospective phase I/II study we evaluated the feasability and efficacy of leucocyte transfusions (LT) in patients with malignancies or haematological disorders who were suffering from severe bacterial or fungal infection during therapy-related bone marrow aplasia. 30 patients with severe neutropenia and clinical signs of life-threatening sepsis not responding to adequate treatment, received LT from rhG-CSF-stimulated family donors or from prednisolone-primed volunteers. A total of 301 LT were administered. The median number of LT per patient was seven (range three to 65), the median duration of LT treatment was 8 d (range 2-35). The white cell count (WBC), absolute neutrophil count (ANC) and lymphocyte count of the concentrates from rhG-CSF-stimulated donors were significantly higher than those from prednisolone-primed volunteers (P = 0.0001). Despite the critical condition of the patients, LT were generally well tolerated. Only 39 (12.9%) LT were associated with adverse reactions. The transfusion of leucocytes collected by continuous flow leukapheresis from both rhG-CSF and prednisolone stimulated donors resulted in a measurable increment of the peripheral leucocyte and ANC counts in our patients. On day 100 after the first LT, 20/30 patients were alive with complete clearance of the infection. PMID- 10468858 TI - Hepatosplenic candidiasis in patients with acute leukaemia. AB - A retrospective study of 23 patients with acute leukaemia and hepatosplenic candidiasis (HSC) was conducted to evaluate clinical treatment characteristics in terms of amount and duration of antifungal agents and to assess treatment outcome. Patients were admitted to two major tertiary care centres between 1990 and 1998. The diagnosis of HSC was based on clinical, blood cultures, histologic and imaging studies. Patients were treated with amphotericin B without interruption of the planned chemotherapy regimens. Serial magnetic resonance imaging (MRI) studies were the main tool for following patients' response and activity of the fungal lesions in conjunction with clinical and laboratory parameters. Treatment with amphotericin B was continued until resolution of all clinical symptoms and signs attributable to HSC, obtaining negative blood cultures and the appearance of at least healed lesions on MRI. Amphotericin B was discontinued in four patients because of severe nephrotoxicity (two patients), or continuous fever and persistent fungal lesions on MRI (two patients). Amphotericin B lipid complex (ABELCET) was successfully used as salvage therapy for these refractory patients. Four patients died with evidence of HSC despite treatment and supportive measures. The response rate for treatment of HSC was 82%. The mean total dose of amphotericin B including empirical treatment was 4 g and the median duration of treatment for responding patients was 112 d. The median number of days of anti- fungal treatment before the disappearance of fever was 19 d. Our results confirmed the need for protracted courses of antifungal agents for the successful eradication of HSC. Chemotherapy for the underlying disorder should not be interrupted or delayed in order to treat HSC. PMID- 10468859 TI - Myelodysplastic syndrome in a patient with adult T-cell leukaemia. AB - A 53-year-old female who developed myelodysplastic syndrome (MDS) after chemotherapy for adult T-cell leukaemia (ATL) is described. The latent period of therapy-related MDS (t-MDS) from the time of diagnosis of ATL was approximately 35 months. Cytogenetic analysis of the bone marrow cells at the time of diagnosis of t-MDS revealed a clonal abnormality; 46,XX,add(7)(p13), der(17)t(3;17)(p11;p13). Although monoclonal integration of human T lymphotropic virus type I (HTLV-I) proviral DNA was detected in the peripheral blood lymphocytes at ATL diagnosis, bone marrow cells at t-MDS diagnosis did not show monoclonal integration of HTLV-I. To our knowledge, this is the first report of t MDS associated with ATL. PMID- 10468860 TI - Mutational analysis of the DNA mismatch repair gene hMLH1 in myeloid leukaemias. AB - Mutations of the DNA mismatch repair (MMR) gene hMLH1 have recently been linked to the development of some hereditary and sporadic cancers which frequently display widespread microsatellite instability (MSI). Conflicting results regarding the extent of MSI in myeloid leukaemias prompted us to perform mutational analysis of all 19 exons of the hMLH1 gene by polymerase chain reaction-single-stranded conformation polymorphism (PCR-SSCP) and sequence analysis in a total of 133 patients with acute and chronic myeloid leukaemia. Apart from one exonic and one intronic polymorphism, no mutations were detected in any of the samples indicating that the major MMR gene hMLH1 is not involved in the pathogenesis or progression of myeloid malignancies. PMID- 10468861 TI - Fas ligand-induced apoptosis of hepatocytes in natural killer cell leukaemia. AB - Neoplastic natural killer (NK) cells overexpress Fas ligand (FasL), which may cause damage of Fas-bearing tissues. We report a patient with NK cell leukaemia who developed liver injury after pharyngitis. The NK leukaemic cells expressed functional FasL. In addition to soluble FasL, serum levels of interleukin-6 and interferon-gamma were increased dramatically when liver injury was aggravated. Moreover, hepatocytes expressed Fas and apoptotic hepatocytes were detected in the portal areas. These findings are consistent with the notion that inflammatory cytokines enhance the sensitivity to FasL and trigger apoptosis of hepatocytes in NK cell malignancies. PMID- 10468862 TI - Monoclonal Epstein-Barr virus-related lymphoproliferative disorder following adult acute lymphoblastic leukaemia. AB - A 31-year-old patient in remission of acute lymphoblastic leukaemia (ALL), receiving oral maintenance chemotherapy (6-mercaptopurine, methotrexate (MTX), cyclophosphamide), developed a monoclonal, Epstein-Barr virus (EBV)-related lymphoproliferative disorder (LPD). Treatment consisted of excisional biopsy and the discontinuation of maintenance chemotherapy. To our knowledge, this is the first such report in an adult. The histological similarity to previous reports of 'lymphomatoid granulomatosis' following paediatric ALL suggests that they are the same disease. MTX may play a central role in the development of LPD in this setting. Although it is a rare complication of ALL, EBV-related LPD should be considered in patients who develop lymphadenopathy. PMID- 10468863 TI - P53 deletion is not a frequent event in multiple myeloma. AB - Recently a high incidence of interstitial deletion of the P53 locus has been reported in multiple myeloma (MM) patients. Considering the importance of such an event, we analysed 79 patients with advanced-stage disease using fluorescence in situ hybridization (FISH). Strikingly, we found only 7/79 patients with a P53 deletion. In order to rule out any differences in probe selection, we reanalysed all the patients with the same probe as that used in a previous study, and confirmed the low incidence of P53 deletion (7/79, 9%). The only explanation is a difference in hybridization efficiency. Since hybridization is far less efficient on malignant plasma cells than on other bone marrow cells we suggest that this poor hybridization efficiency may lead to a false P53 deletion. PMID- 10468865 TI - Incompatibility for CD31 and human platelet antigens and acute graft-versus-host disease after bone marrow transplantation. AB - Bone marrow transplantation (BMT) is often complicated by acute graft-versus-host disease (aGVHD). In patients transplanted with an HLA-matched donor the occurrence of this complication is believed to be favoured by disparities at the minor histocompatibility antigens (mHA). However, few of these polymorphic molecules have been identified. We sought to determine whether donor/recipient incompatibility for HPA-1, HPA-2, HPA-3, HPA-5 or CD31 (codon 125) antigens represented a risk factor for aGVHD and genotyped these antigens in 70 bone marrow donors and their HLA-identical recipients. All patients were children who received BMT for haematological malignancies at a single institution according to well-defined therapy protocols. Statistical analysis showed that incompatibility for CD31 (codon 125) was a risk factor for grade II-IV aGVHD in the overall patient population, whereas HPA-3 incompatibility predicted aGVHD occurrence in HLA-A2 patients only. The magnitude of the aGVHD risk was directly related to the number of HPA/CD31 incompatibilities. No correlation was found between non identity for HPA/CD31 and aGVHD. Since incompatibility but not non-identity for CD31 or HPA-3 was a risk factor for aGVHD, we suggest that allelic variants of these molecules can serve as mHA in BMT recipients from HLA-identical donors. PMID- 10468864 TI - Additional translocation (8;21)(q22;q22) in a patient with Philadelphia-positive chronic myelogenous leukaemia in the blastic phase. AB - We report a case of Philadelphia-positive chronic myelogenous leukaemia in blastic phase with the additional translocation (8;21)(q22;q22), which is frequent in acute myeloid leukaemia but not in chronic myelogenous leukaemia. The t(8;21) was not detected in the chronic phase, and was the only additional chromosomal anomaly in the blastic clone. Reverse transcription-polymerase chain reaction revealed the AML1/ETO fusion transcript in the cells of blastic phase but not in those of chronic phase. Regarding t(9;22), the breakpoint on chromosome 22 occurred in the mu-BCR region of the BCR gene, resulting in hybrid BCR/ABL mRNA with an e19a2 junction. Our findings provided molecular evidence that t(8;21) can occur as an additional genetic change in Philadelphia-positive chronic myelogenous leukaemia. PMID- 10468866 TI - Phase III efficacy study of interleukin-3 after autologous bone marrow transplantation in patients with malignant lymphoma. AB - We evaluated the efficacy of recombinant human interleukin-3 (rhIL-3) in reducing the number of platelet transfusions and major infections after autologous bone marrow transplantation (ABMT) in patients with malignant lymphoma. 198 patients with non-Hodgkin's lymphoma (NHL, n = 111) and Hodgkin's disease (HD, n = 87) were randomized to receive rhIL-3 10 microgram/kg/d (n = 130) or placebo (n = 68) for a maximum of 28 d after ABMT. Several well-known conditioning regimens were used. From day 1 after ABMT patients were treated with placebo or rhIL-3 at a dose of 10 microgram/kg/d by continuous i.v. infusion for 7 d and then by s.c. administration for 21 d or until platelet (50 x 109/l) and neutrophil (0.5 x 109/l) recovery had occurred. Treatment was completed in 54% of the patients in the rhIL-3 group versus 75% in the placebo group (P < 0.004). Adverse events were the main reason for premature discontinuation in the IL-3 group (23% IL-3 v 5% placebo). The median number of platelet transfusions was not significantly different between the IL-3 group and the placebo group (8.0 IL-3 v 6.0 placebo, P = 0.09). Platelet engraftment (>/= 20 x 109/l) was not significantly faster in the IL-3 group (28 d in the IL-3 and 27 d in the placebo group, P = 0.06) and the incidence of haemorrhagic complications was similar in both groups. In patients receiving the full intended dose of rhIL-3, platelet engraftment to >/= 20 x 109/l was delayed (P = 0.007). The median time to neutrophil engraftment was 23 d in the IL-3 and 25 d for the placebo group (P = 0.39). There was no difference in the incidence of major infections. We conclude that treatment with IL-3 has no clinical benefit in patients receiving ABMT for malignant lymphoma. PMID- 10468867 TI - The relationship between monoclonal myeloma precursor B cells in the peripheral blood stem cell harvests and the clinical response of multiple myeloma patients. AB - The aim of this study was to determine the presence of monoclonal myeloma precursor B cells in peripheral blood stem cell harvests and to investigate their role in the clinical outcome of multiple myeloma patients. A total of 39 multiple myeloma patients were treated with a sequential therapy including double high dose melphalan therapy followed by a double transplant procedure. The apheresis products for the second transplant were purged using a panel of four or five different mouse monoclonal antibodies against B-cell antigens (CD10, CD19, CD20, CD22 and CD37). In 19/39 patients a tumour-specific CDR III signal was identified in the diagnostic bone marrow. Gene scan analysis after CDR III PCR of the magnetic bead isolated B-cell fraction from the apheresis products in these 19 patients revealed three different patterns: 32% of patients had a predominantly monoclonal B-cell population; 63% of patients had an identifiable monoclonal signal within an oligoclonal B-cell population. In only 1/19 patients were no monoclonal B cells identified in the B-cell population of the apheresis product. A correlation between the clonal pattern and the clinical response after sequential chemotherapy was found. Patients with a predominance of monoclonal myeloma or myeloma precursor B cells had an early relapse or achieved a minimal response or a partial remission. Patients with an oligo- and/or polyclonal pattern achieved a high percentage of partial as well as complete remissions. PMID- 10468868 TI - Clonal immunoglobulin light chain variable region germline gene use in AL amyloidosis: association with dominant amyloid-related organ involvement and survival after stem cell transplantation. AB - AL (primary or immunoglobulin light chain) amyloidosis (AL) differs from myeloma per se in that tissue deposits of amyloid are found, typically in association with small numbers of clonal plasma cells producing lambda light chains, and also in that AL patients typically present with a predominantly dysfunctional organ system. This constellation of features - fibrillar deposits comprised of light chains, lambda light chain predominance, and organ-system tropism and dysfunction - remains unexplained. Select patients with AL respond to haemopoietic stem cell transplantation (SCT) with clinical improvement and extended survival, particularly those who do not have cardiac involvement. In order to ascertain whether the organ-system tropism of AL was associated with immunoglublin light chain variable region (Ig VL) germline gene utilization, we attempted to clone, sequence and assign germline donors to the clonal Ig VL genes of 62 AL patients, all of whom were treated with SCT. We succeeded in 39 cases, identifying clonal AL genes derived from donors of the lambdaI (n = 10), lambdaII (n = 5), lambdaIII (n = 6), lambdaVI (n = 11) and KI (n = 7) subtypes. The majority of the donors (IGLV6S1, DPL5, DPL2, DPL23 and LFVK431) were genes that appear in the expressed repertoire <5% of the time, suggesting an intrinsic propensity to form amyloid under certain conditions. Patients whose clones derived from the lambdaVI IGLV6S1 donor uniformly presented with dominant renal involvement while those with other Vlambda or unknown donors often had dominant cardiac or other organ involvement, particularly patients whose clones derived from the lambdaI DPL2 donor. In addition, both early (<3 months) and overall post-SCT survival were significantly better in lambdaVI IGLV6S1 patients compared to patients with other Vlambda donors. These findings indicate that there are important associations in AL amyloidosis among Ig VL gene utilization, organ-system tropism and post-SCT survival. PMID- 10468869 TI - Soluble transferrin receptor as a potential determinant of iron loading in congenital anaemias due to ineffective erythropoiesis. AB - Congenital anaemias due to ineffective erythropoiesis may be associated with excessive iron absorption and progressive iron loading. We investigated whether the soluble transferrin receptor (TfR) level was related to the degree of iron overload in 20 patients with thalassaemia intermedia, six patients with congenital dyserythropoietic anaemia type II (CDA II) and four patients with X linked congenital sideroblastic anaemia (XLSA). All but two patients had increased serum ferritin levels (median 601 microgram/l, range 105-2855 microgram/l). Multiple regression analysis showed that 62% (P < 0.0001) of the variation in serum ferritin was explained by age and by changes in soluble TfR. PMID- 10468870 TI - Defective functional activity and accelerated apoptosis in neutrophils from children with cancer are differentially corrected by granulocyte and granulocyte macrophage colony stimulating factors in vitro. AB - We have previously shown that polymorphonuclear leucocytes (PMN) harvested from children with cancer and exposed to chemotherapy exhibit defective bactericidal activities against both Gram-positive and Gram-negative microorganisms as well as accelerated apoptosis. In this study, PMN from children with cancer were evaluated to compare in vitro the corrective effects of the two myeloid colony stimulating factors G-CSF and GM-CSF on these defective pathways. Both G-CSF and GM-CSF were able to increase the defective bactericidal activities against S. aureus and E. coli. However, GM-CSF was consistently superior to G-CSF in correcting PMN microbicidal activity; this correction was incomplete since it did not reach the level observed in normal PMN exposed to GM-CSF. The accelerated apoptosis of PMN was not affected by G-CSF. In contrast, GM-CSF significantly prolonged the survival of the PMN although it did not reach the level of survival observed with normal PMN exposed to GM-CSF. These observations were consistent with other studies indicating that in PMN, microbicidal activities and apoptosis are differentially sensitive to the myeloid growth factors G-CSF and GM-CSF. PMID- 10468871 TI - Measurement of endogenous and exogenous alpha-granular platelet proteins in patients with immune and nonimmune thrombocytopenia. AB - Idiopathic thrombocytopenic purpura (ITP) is caused by antiplatelet antibodies and is characterized by increased platelet destruction and elevated levels of IgG (platelet-associated IgG, PAIgG). Nonimmune thrombocytopenic patients also have elevated levels of PAIgG. In this study we investigated two possible biological explanations for the increased levels of PAIgG in these patients. The first hypothesis suggests that a thrombocytopenic stress causes increased thrombocytopoiesis with increased numbers and content of the platelet alpha granules. The second hypothesis is that for uncertain reasons (immunological or cytokine) there is increased absorption of plasma proteins by either megakaryocytes or by the platelets themselves. To address this issue, we compared the level of megakaryocyte synthesized alpha granular proteins [platelet factor 4 (PF4) and beta-thromboglobulin (beta-TG)] to plasma-absorbed alpha granular proteins (albumin, IgG and fibrinogen) in patients with immune (n = 39) and nonimmune (n = 60) thrombocytopenias. Plasma-absorbed alpha-granular proteins were elevated in both immune and nonimmune thrombocytopenia with no increase in megakaryocyte synthesized alpha-granular proteins. These plasma-derived protein elevations were not attributable to elevated mean platelet volumes or elevated plasma concentrations of the respective protein. We hypothesize that the increased IgG in these platelets is not the result of production of larger platelets, but reflects a selective increase in the endocytosis of plasma absorbed alpha-granular proteins at the megakaryocyte and/or platelet level. PMID- 10468872 TI - The GPIa C807T dimorphism associated with platelet collagen receptor density is not a risk factor for myocardial infarction. AB - The platelet collagen receptor, GPIa/IIa, is an important mediator of platelet adhesion to fibrillar collagens at sites of vascular injury. Recently, a dimorphism at nucleotide 807 of the GPIa cDNA (TTC/TTT in codon 224) was shown to be associated with variation in GPIa/IIa receptor density on the platelet surface. We conducted a case-control study to determine if the 807T allele, linked with increased GPIa/IIa density, contributed to risk of myocardial infarction (MI). DNA from 546 acute MI cases and 507 controls, all aged <75 years, was genotyped for the C807T dimorphism using the TaqManTM system of allelic discrimination. The allelic odds ratio (OR) for MI in the complete cohort was 0.88 (95% CI 0.74-1.05, P = 0.17), indicating that the 807T allele was not associated with an increased risk of MI. There was also no increased risk of MI associated with the homozygous 807TT (P = 0.22) or heterozygous 807CT (P = 0.24) genotypes or for carriers of the 807T allele in any cohort subgroup analysed. We conclude that the GPIa 807T allele is not a risk factor for MI in our population either alone or in combination with other major cardiovascular risk factors. PMID- 10468873 TI - Therapeutic monitoring of von Willebrand disease: interest and limits of a platelet function analyser at high shear rates. AB - We have evaluated the position of the Platelet Function Analyzer PFA-100TM in the management of 41 patients with von Willebrand disease (VWD) receiving either desmopressin (23 patients with type 1, five with type 2M, three with type 2A and three with type 2B) or von Willebrand factor (VWF) concentrates (four patients with type 3, two with type 2M 'type B', two with type 2A and one type 1 'platelet low'). In all patients the following were studied before and 30 min after infusion of desmopressin and/or VWF concentrates: VWF ristocetin cofactor activity (VWFRCo), bleeding time (BT) and closure time with the PFA-100 using ADP (CT-ADP) as well as epinephrine (CT-Epi) cartridges. After the infusion of desmopressin, the CT was modified in the same way as the VWFRCo levels, being always normalized in patients with type 1 and not constantly corrected in those with type 2. Thus, our results indicated that the measurement of the CT enabled a quick and accurate evaluation of the response to desmopressin which, in fact, measured the releasable VWF cellular compartment containing the highly multimerized forms of VWF. For patients with type 2 or 3 VWD who were non responsive to desmopressin, VWF concentrates corrected the VWFRCo defect but not the CT as none of these patients had a normal platelet VWF content and the VWF concentrates did not contain the ultralarge VWF multimers. In conclusion, the very high shear conditions in the PFA-100 make it very sensitive to the contribution of platelet VWF and to the ultralarge VWF multimers, indicating that the evaluation of the CT is a very simple and rapid tool to discriminate between good and non-responders to desmopressin. PMID- 10468874 TI - Effect of factor VIII concentrate on leucocyte cytokine production: characterization of TGF-beta as an immunomodulatory component in plasma-derived factor VIII concentrate. AB - Clinical and subclinical immunological abnormalities have been reported in HIV seronegative haemophiliacs. The mechanisms by which these abnormalities arise remain unclear. As cytokines are important biological response modifiers, the effect of a FVIII concentrate on production of a range of cytokines, by a variety of cells, was investigated. A whole blood technique was used and the in vitro modulation of cytokine synthesis by an intermediate-purity plasma-derived factor VIII (pdFVIII) concentrate was analysed using multiparameter flow cytometry. In cell cultures exposed to pdFVIII, T cells showed reduced production of TNF-alpha, IL-2 and IFN-gamma; monocytes showed reduced production of TNF-alpha, IL-1alpha, IL-1beta, IL-6, IL-8 and IL-12 but an increase in IL-10 synthesis; IFN-gamma synthesis by NK cells was reduced. All changes in cytokine synthesis and the reduction in cell surface expression of CD69, a signal transduction molecule contributing to both cytokine and cytokine receptor synthesis, were in a dose dependent manner in cultures exposed to FVIII concentrate. These changes were characteristic of TGF-beta. Addition of anti-TGF-beta to FVIII reduced these changes in T-cell cytokine production, suggesting TGF-beta may be an important immunomodulatory agent in the pdFVIII concentrate. The Th2 cytokine bias shown in the presence of pdFVIII concentrate, in vitro, may explain the increase in rates of certain types of infections reported in these patients, which require Th1 cytokine production for an effective response. PMID- 10468875 TI - Substitution of Arg527 and Arg531 in factor VIII associated with mild haemophilia A: characterization in terms of subunit interaction and cofactor function. AB - The functional defect caused by substitution of Arg527 (--> Trp) and Arg531 (--> Gly, His) in factor VIII (FVIII), was explored by employing FVIII derived from patient plasma and recombinant FVIII variants. Mutation of these residues is associated with mild haemophilia A. For both FVIII-R527W and FVIII-R531H, activity was lower than antigen, indicating a functional defect for both variants. In contrast to FVIII-R527W, the amount of FVIII-R531H heterodimer present in plasma was reduced compared to heavy and light chain levels. Factor X (FX) activation experiments employing recombinant FVIII-R531G revealed that the activated FVIII-R531G heterotrimer was less stable than normal FVIIIa, apparently due to rapid dissociation of the A2 domain. These findings suggest that Arg531 is involved in maintaining the stability of both the heterodimer and the activated FVIII heterotrimer. Recombinant FVIII-R527W displayed reduced stimulation of FX activation, suggesting a defect in interaction with factor IXa (FIXa). The contribution of Arg527 in the interaction with FIXa was supported by the observation that FVIII-derived synthetic peptide Tyr511-Leu530 was able to inhibit FX activation and that this inhibition could be overcome by addition of increasing concentrations of FIXa. Furthermore, in the three-dimensional FVIII model residues Val517-Arg527 are located near the FIXa binding site Ser558 Gln565. Therefore we propose that Arg527 is part of an extended FIXa binding site, comprising residues Ser558-Gln565 and Val517-Arg527. PMID- 10468876 TI - Lipid composition of seven APTT reagents in relation to heparin sensitivity. AB - The phospholipid content of different activated partial thromboplastin time (APTT) reagents was determined and compared to heparin sensitivity. The seven reagents included were those most widely used amongst participants of the U.K. National External Quality Assessment Scheme (NEQAS) at the time of study. Heparin sensitivity was assessed using the APTT ratios obtained by more than 300 NEQAS participants on five plasmas prepared from patients receiving unfractionated heparin. The concentrations of three neutral lipids and six phospholipids present in the seven APTT reagents were determined by high-performance thin-layer chromatography (HPTLC) and densitometry. Both the concentrations and the relative percentages of individual phospholipid components varied markedly between reagents. The total phospholipid concentration included a 12-fold range from 16 to 205 microgram/ml. Phosphatidylserine (PS) was completely lacking from one reagent prepared from vegetable material and ranged from 3 to 22 microgram/ml in the other six reagents containing extracts from animal tissue. The concentration of phosphatidylcholine ranged from 3 to 109 microgram/ml. There was no demonstrable relationship between the concentration of any individual lipid components and heparin sensitivity. However, the relative percentage phospholipid composition was important since a lower % of PS or phosphatidylinositol (PI) correlated with increasing heparin sensitivity. PMID- 10468877 TI - A family with hereditary factor X deficiency with a point mutation Gla32 to Gln in the Gla domain (factor X Tokyo). AB - We report a new family with hereditary factor X deficiency. The propositus had a markedly prolonged prothrombin time, a mild prolongation of activated partial thromboplastin time and a clotting time activated by Russell's viper venom. Factor X activity in plasma was 3 u/dl (normal range 56-138 u/dl). Factor X antigen level was 61 u/dl. Molecular analysis revealed a homozygous mutation, Glu (GAG) to Gln (CAG) at residue 32 which normally undergoes gamma-carboxylation within the gamma-carboxyglutamic acid rich domain. The genotypes of family members completely correlated with their factor X activities. It is suggested that the Glu32 to Gln mutation is the molecular basis for the abnormal factor X in this family. PMID- 10468878 TI - Is there a correlation between degree of splenomegaly, symptoms and hypersplenism? A study of 218 patients with Gaucher disease. AB - Despite the prevalence of splenomegaly as a sign in many disorders, there have been no studies that correlate the degree of organomegaly with the symptoms generally ascribed to splenic enlargement. The degree of splenomegaly was compared with five overt symptoms of mechanical displacement, i.e. chronic abdominal pain, abdominal discomfort, early satiety, pain while lying on the side, or attacks of acute (colicky) left upper quadrant pains. We have also employed splenomegaly as seen in Gaucher disease as a paradigm to determine whether there is a correlation between the degree of splenomegaly and the parameters of hypersplenism. Although there was a statistically significant correlation between degree of splenomegaly and blood counts, this proved to be clinically negligible. Surprisingly, there was also no correlation between degree of splenomegaly and any of symptoms investigated. PMID- 10468879 TI - Production and characterization of anti-kell monoclonal antibodies using transfected cells as the immunogen. AB - Monoclonal antibodies (Mabs) to blood group antigens are valuable as diagnostic reagents for typing red blood cells (RBCs) in the clinical setting, and for structure-function studies of proteins. Here, we report a powerful system that enabled us to produce Mabs to blood group antigens. A murine erythroleukaemia (MEL) cell line expressing Kell protein, a transmembrane glycoprotein that carries a number of clinically relevant antigens, was used as a novel immunogen. Mabs with different specificities to the Kell protein were produced from a single mouse fusion: an anti-Jsb (MIMA-8), and two antibodies (MIMA-9 and MIMA-10) with novel specificities, that reacted with RBCs with the common Kell phenotype but not with RBCs with K+k- or Kp(a+b-) or K0 phenotypes. The non-reactivity with both K+k- or Kp(a+b-) RBCs implied that the epitope was influenced by the molecular changes associated with an absence of the k or Kpb antigens. MIMA-8 is the first example of a Mab anti-Jsb and was used in the clinical laboratory for screening donor RBCs for Js(b-) blood and for typing RBCs from patients even when the RBCs were coated with anti-IgG as is the case in autoimmune haemolytic anaemia. Heavy and light chain variable regions of MIMA-8 were cloned and the sequence is given. This study illustrates the potential of this novel immunization approach for making monoclonal antibodies to blood group antigens. PMID- 10468880 TI - A prospective study of routine antenatal enzyme antibody screening demonstrates lack of clinical value in predicting haemolytic disease of the newborn. AB - A prospective study of 7065 consecutive new pregnancies identified 230 with a positive screen, of which 27% (62/230) were 'enzyme-only' antibodies. 32 of these (52%) were potentially clinically important and were all of Rh specificity: 22 anti-E, seven anti-Cw, two anti-D and one anti-c. However, only three of these enzyme-only antibodies (one anti-D, one anti-c and one anti-E) became reactive by the indirect antiglobulin test (IAT) during the course of pregnancy, and all were detected in the routine 34-36-week maternal sample. No babies were affected, and we reaffirm that routine antibody screening by enzyme techniques is unnecessary. PMID- 10468882 TI - The management of 'low-risk' and 'intermediate-risk' patients with primary thrombocythaemia. MPD (UK) Study Group. PMID- 10468881 TI - Functional expression of TRAIL by lymphoid and myeloid tumour cells. AB - TRAIL is a potent death protein that favours the killing of various types of cancer cells to normal cells, but under the right conditions TRAIL can also kill activated human T cells. TRAIL mRNA is widely expressed by normal cells but its expression by primary tumour cells is not known. In this study, primary tumour cells of haemopoietic origin constitutively expressed TRAIL mRNA and protein and were capable of inducing the apoptosis of target Jurkat cells in a dose-dependent manner. This killing effect was reversed by anti-TRAIL antibody. The functional expression of TRAIL by lymphoid and myeloid malignant cells raises the possibility of its involvement in tumour cell evasion of immunosurveillance, and could be related to spontaneous tumour cell death and necrosis. PMID- 10468883 TI - THE MANAGEMENT OF 'LOW-RISK' AND 'INTERMEDIATE-RISK' PATIENTS WITH PRIMARY THROMBOCYTHAEMIA PMID- 10468884 TI - Does anti-D protect platelets from anti-HPA 1A mediated destruction? PMID- 10468885 TI - Anticipation in familial plasma cell disease. PMID- 10468886 TI - C-kit activating mutation in a neonate with in-utero presentation of systemic mastocytosis associated with myeloproliferative disorder. PMID- 10468887 TI - Helicobacter pylori infection on the risk of stomach cancer and chronic atrophic gastritis. AB - Helicobacter pylori infection is associated with gastric adenocarcinoma. However, the mechanisms of this interaction are still unclear. This study was conducted to explore the effects of H. pylori infection on early and late stage gastric carcinogenesis. This study included 134 patients with adenocarcinoma of the stomach (ACS), 67 patients with chronic atrophic gastritis (CAG), and 65 normal controls recruited at Memorial Sloan-Kettering Cancer Center (MSKCC) from November 1, 1992 to November 1, 1994. Epidemiologic data were collected by a modified National Cancer Institute Health Habits History Questionnaire. H. pylori infection was diagnosed by pathological evaluation. Risk factors were analyzed using logistic regression. The odds ratio (OR) associated with H. pylori infection was 10.4 [95% confidence interval (CI): 2.6-41.6] for CAG and 11.2 (95% CI: 2.5-50.3) for gastric cancer in comparison with normal controls, with adjustment for pack-years of smoking, alcohol drinking, body mass index, total caloric intake, dietary fat and fiber intake, and Barrett's esophagus. But H. pylori infection was not associated with risk of stomach cancer when patients with stomach cancer were compared with patients with CAG (OR = 0.6, 95% CI: 0.3 1.3) after controlling for potential confounding variables. This association was persistent when only patients with both gastric cancer and chronic gastritis were considered as cases and patients with CAG were considered as controls (OR = 0.7, 95% CI: 0.3-2.0) in the multivariate analysis. Our results suggest that H. pylori infection may be involved in the early stage of development of CAG, but not in the development of stomach cancer from CAG, and indicate that strategies for prevention of stomach cancer should target the early stage to eliminate H. pylori infection in high-risk populations. PMID- 10468888 TI - Secondary tumors in bone sarcomas after treatment with chemotherapy. AB - New oncologic treatments have improved survival in osteosarcoma and Ewing's sarcoma. However, these treatments may cause secondary malignancies after radiotherapy. This study evaluated the incidence of secondary malignancies after neoadjuvant chemotherapy. Between April 1972 and December 1990, 518 osteosarcoma and 299 Ewing's sarcoma patients entered neoadjuvant chemotherapy protocols. Follow-up records of all patients were analyzed and malignant tumors were reported. Nine patients developed another malignancy, including 5 leukemias, 1 astrocytoma, 1 liposarcoma, 1 parotid, and 1 breast carcinoma. Four leukemias were found in patients treated for osteosarcoma with chemotherapy, but not radiotherapy. Only one leukemia developed after Ewing's sarcoma treated with chemotherapy and radiotherapy. The incidence of leukemias is high, while the other tumors can be explained as unrelated cases. Incidence densities for leukemia were calculated for both groups of patients. Treated osteosarcoma patients seem to have a predisposition to develop leukemias, but whether this is chemotherapy induced needs to be investigated. PMID- 10468889 TI - Differential expression of and responsiveness to transforming growth factor-beta (TGF-beta) isoforms in hormone-dependent and independent lines of mouse mammary tumors. AB - Transforming growth factor-beta2 (TGF-beta2) and -beta3 mRNA expressions were studied in ductal hormone-dependent (HD) and -independent (HI) in vivo lines of the medroxyprogesterone acetate (MPA)-induced mammary tumor model in Balb/c mice. MPA treatment of HD tumors induced a significant decrease in TGF-beta2 and -beta3 mRNA levels. Progression to an HI phenotype of ductal tumors was associated with reduced TGF-beta2 and -beta3 expressions, as compared with their HD counterparts. Exogenously added TGF-beta1, -beta2, and -beta3 (1 ng/ml) inhibited the proliferation of primary cultures of epithelial cells from ductal HD and HI tumors. In addition, TGF-beta expression and effects were studied in the other type of MPA-induced mammary tumors, which are of lobular origin and lack steroid hormone receptors and evidence an HI behavior. These lobular HI lines showed TGF beta2 levels similar to those found in HD lines growing in MPA-treated mice. In contrast, TGF-beta3 mRNA levels were 12- to 20-fold higher than in HD tumors. Primary cultures of lobular HI epithelial cells required either TGF-beta concentrations of 10 ng/ml to show an inhibitory response, or were completely resistant to TGF-beta inhibition. Studies of the molecular mechanisms involved in reduction or loss of TGF-beta responsiveness in lobular HI tumors showed that cell surface type II TGF-beta receptor levels were lower in these tumors than those present in HD tumors. Our results support the hypothesis that TGF-beta could play a role as an autocrine growth inhibitor in HD and HI ductal tumors. Autonomous growth of lobular HI tumors could be favored by undetectable or low TGF-beta1 and -beta2 expressions and by reduced or lost sensitivity of epithelial cells to TGF-beta's antiproliferative effects. However, the extremely high levels of TGF-beta3 expression in lobular HI tumors, in spite of reduced sensitivity to TGF-beta3 inhibitory growth effect in tumor epithelial cells, suggest a net positive role for TGF-beta3 in these tumors. PMID- 10468890 TI - Development and characterization of melanoma cell lines established by fine needle aspiration biopsy: advances in the monitoring of patients with metastatic melanoma. AB - The establishment of melanoma cell lines from fine-needle aspiration biopsies (FNAB) has allowed for an enhanced understanding of the complex interactions that occur between T cells and tumor cells. The technique of FNAB offers the advantage of providing a sequential analysis of the same tumor nodules throughout treatment. The expression of melanoma antigens (MAs) was assessed in fresh melanoma FNAB samples and from tumor cell lines derived from these samples using several different approaches. Cytospin preparations of freshly isolated tumor cell explants were analyzed by immunocytochemistry (ICC), while the daughter cell line was analyzed by fluorescent activated cell sorting (FACS) analysis, and semiquantitative and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR, qRT-PCR). As assessed by these methods, the level of MA expression by the original tumor cell explants correlated with the expression in established in vitro cell lines. Molecular analysis of the established cell lines utilizing PCR technology improved the sensitivity of detection of MA expression. Thus FNAB of melanoma is an efficient and effective method of tissue procurement, capable of generating, sequentially and from the same lesion, fresh tumor cells, tumor infiltrating lymphocytes (TIL), and long-term melanoma cell lines. PMID- 10468891 TI - Role of serial tumor markers in the surveillance for recurrence in endometrial cancer. AB - The aim of this study is to evaluate the usefulness of tumor markers in the follow-up of patients with endometrial cancer. The sera of 23 patients with elevated pretreatment CA125, CA15.3, and CA19.9 were collected at each follow-up visit and analyzed. Eleven patients had normal posttreatment levels and none of them developed recurrence. Twelve patients had one or more increased tumor markers, 7 (58.3%) of them developed recurrence. Among these seven patients, six had CA125 elevated by at least 10-fold and one had elevated CA19.9. The median lead time between elevation of tumor markers and clinical evidence of recurrence was 6 months. Hence posttreatment elevation of CA125 is a useful predictor for recurrence in patients with elevated pretreatment levels. PMID- 10468892 TI - Image cytometric DNA analysis and proliferating cell nuclear antigen (PCNA) expression in transitional cell carcinoma of the bladder. AB - The proliferation rate as determined by Proliferating Cell Nuclear Antigen (PCNA) immunostaining and the DNA ploidy status as measured by static cytometry were studied in 70 transitional cell carcinomas of the bladder (TCCB) in relation to grade, stage, and recurrence. The follow-up period was 2 years. A significant difference was observed in PCNA expression among grades I, II, and III (P < 0.02), between superficial (pTa-pT1) and invasive (pT2-pT4) tumors (p < 0. 04), between recurring and non-recurring tumors (p < 0.001), and between tumors of the same grade with and without recurrence (p < 0. 05). A significant difference was also found in the ploidy pattern among grades I, II, and III (p = 0.002), and between superficial and invasive (p = 0.02) and recurring and non-recurring tumors (p < 0. 01). Finally, the recurrence status seems to be strongly influenced by the proliferation rate and ploidy of TCCB (p < 0.001). Our results suggest that the above-studied parameters may offer useful information on the biological behavior of TCCB. PMID- 10468893 TI - Responsiveness of human prostate carcinoma bone tumors to interleukin-2 therapy in a mouse xenograft tumor model. AB - We have tested an immunotherapy approach for the treatment of metastatic prostate carcinoma using a bone tumor model. Human PC-3 prostate carcinoma tumor cells were heterotransplanted into the femur cavity of athymic Balb/c nude mice. Tumor cells replaced marrow cells in the bone cavity, invaded adjacent bone and muscle tissues, and formed a palpable tumor at the hip joint. PC-3/IF cell lines, generated from bone tumors by serial in vivo passages, grew with faster kinetics in the femur and metastasized to inguinal lymph nodes. Established tumors were treated with systemic interleukin-2 (IL-2) injections. IL-2 significantly inhibited the formation of palpable tumors and prolonged mouse survival at nontoxic low doses. Histologically IL-2 caused vascular damage and infiltration of polymorphonuclear cells and lymphocytes in the tumor as well as necrotic areas with apoptotic cells. These findings suggest destruction of tumor cells by systemic IL-2 therapy and IL-2 responsiveness of prostate carcinoma bone tumors. PMID- 10468894 TI - Granulocyte apheresis as a possible new approach in cancer therapy: A pilot study involving two cases. AB - Patients with advanced cancer often develop immunodeficiency which may be associated with granulocytosis. The granulocytes have the potential to deplete cytotoxic T cells, resulting in accelerated tumor growth and metastasis. To study the elimination of excess granulocytes using granulocyte apheresis in patients with elevated granulocyte to lymphocyte ratios, 2 patients with recurrent metastatic tumors, were selected. Granulocyte apheresis was performed by extracorporeal vein-to-vein circulation with the G-1 granulocyte and monocyte/macrophage apheresis column filled with cellulose acetate beads, each 2 mm in diameter to adsorb granulocytes and monocytes/macrophages. The patients received 1 or 2 apheresis of 30 to 50 min duration per week, at a flow rate of 30 50 ml/min, with 15 sessions constituting one therapeutic course. Apheresis markedly reduced tumor size and prolonged patient survival time without causing any serious adverse events. The results of the present study suggest that granulocyte and monocyte/macrophage apheresis may be beneficial in patients with metastasizing tumors. PMID- 10468895 TI - Modulation of O6-methylguanine-DNA methyltransferase-mediated 1-(4-amino-2-methyl 5-Pyrimidinyl)- methyl-3-(2-Chloroethyl)-3-nitrosourea resistance by antisense RNA. AB - Mer+ HeLa S3 tumor cells with high levels of O6-methylguanine-DNA methyltransferase (MGMT) gene expression were transduced by retroviral-mediated MGMT antisense RNA. The MGMT mRNA, MGMT protein, and MGMT activity in transduced cells were only 28.7%, 32.7%, and 39. 1% of that in HeLa S3 cells, respectively. The transduced cells showed more sensitive to ACNU than HeLa S3 cells both in cell survival and in nude mice experiments. Pathologic examination confirmed that transduced grafts were killed by ACNU. Our results suggested that MGMT gene expression could be modulated by retroviral-mediated antisense RNA and that Mer+ tumor drug resistance to ACNU could be reversed by modulation of MGMT gene expression. PMID- 10468896 TI - Colon cancer treatment in rural North and South Carolina. AB - The purpose of this study was to determine the degree to which colon cancer treatment in rural North and South Carolina in 1991 and 1996 conformed to national treatment recommendations. Data came from medical records of colon cancer patients residing in rural North and South Carolina. The National Cancer Institute's Physician Data Query (PDQ) database was used to define state-of-the art care and to categorize receipt of primary and/or adjuvant treatment. Changes in treatment over time, location, and stage and bivariate relationships between treatment and selected covariates were assessed with chi-square and Fisher's exact tests. Regression was used to control for possible interactions between patient and/or disease characteristics and treatment. The majority of colon cancer cases received primary therapy as suggested by the PDQ which was not significantly related to other factors examined. There was variation in provision of adjuvant therapy. Stage III patients received adjuvant therapy significantly more often than did stage II patients (p 2500 ml) and oliguria/hyponatraemia (< 132 mmol/l) were surveyed as well as predictors of postoperative morbidity. RESULTS: 487 patients (31%) developed immediate postoperative hypotonic polyuria, 161 patients (10%) showed prolonged polyuria and 37 patients (2.4%) delayed hyponatraemia. A biphasic (polyuria-hyponatraemia) and triphasic (polyuria hyponatraemia-polyuria) pattern was seen in 53 (3.4%) and 18 (1.1%) patients, respectively. Forty-one patients (2.6%) displayed immediate postoperative (day 1) hyponatraemia. Altogether, 8.4% of patients developed hyponatraemia at some time up to the 10th day postoperative, with symptomatic hyponatraemia in 32 patients (2.1%). Risk analysis showed that patients with Cushing's disease had a fourfold higher risk of polyuria than patients with acromegaly and a 2.8-fold higher risk for postoperative hyponatraemia. Younger age, male sex, and intrasellar expansion were associated with a higher risk of hypotonic polyuria, but this was not considered clinically relevant. CONCLUSIONS: The analysis illustrates that disturbances in osmoregulation resulting in polyuria and pertubations of serum sodium concentration are of very high prevalence and need observation even after selective transsphenoidal surgery for pituitary adenomas, especially in patients with Cushing's disease. PMID- 10468902 TI - Evaluation of free insulin-like growth factor-I measurement on the diagnosis and follow-up treatment of growth hormone-deficient adult patients. AB - OBJECTIVE: Insulin-like growth factor (IGF-I) and IGF binding protein-3 (IGFBP-3) are GH-dependent and their concentrations have been used in the diagnosis of GH deficiency. Recently, the free fraction of IGF-I has received more attention. The aim of the study was to assess the role of free IGF-I in the diagnosis of GH deficiency in adults, and in follow-up during treatment with recombinant human GH (rhGH). DESIGN AND PATIENTS: We studied 24 adult patients with pituitary disease and GH deficiency and 25 matched controls. Nine patients were re-evaluated after 6 months of treatment with rhGH (0.25 U/kg/week). MEASUREMENTS: Serum levels of IGF-I, free IGF-I, IGFBP-3 and IGFBP-1 were measured by immunoradiometric assay. RESULTS: Serum free IGF-I levels were significantly lower in the GH deficient group than in the normal group (mean: 0.84 and 1.32 micrograms/l respectively, P = 0.0009). Furthermore, serum IGF-I levels were also lower (mean: 92.24 and 230.47 micrograms/l respectively, P < 0.0001). 63% of patients had serum IGF-I concentration below the normal range. For free IGF-I, 52% of the GH deficient patients showed levels below the lowest value obtained for the normal group. Seventy-five percent of the patients showed at least one of the two determinations below the normal range. The free-total IGF-I ratio was significantly higher (P = 0.025) in GH deficient group (range: 0.19-21.29, mean: 2.53) than in normal controls (range: 0.2-2.15, mean: 0.6). Regarding IGFBP-3 and IGFBP-1 no differences were observed between the two groups. During rhGH treatment the increase in serum total and free IGF-I and IGFBP-3 paralleled the beneficial effects on body composition. CONCLUSIONS: Free IGF-I may be another useful method for the diagnosis of GH deficiency, particularly if related to total IGF-I concentration. PMID- 10468903 TI - Discrepancy between serum leptin values and total body fat in response to the oral growth hormone secretagogue MK-677. AB - OBJECTIVE: Growth hormone (GH) treatment decreases total body fat while this effect has not yet been documented for the oral GH secretagogue MK-677. In the present study, the effects of MK-677 treatment on serum levels of leptin, thyroid hormones and testosterone were determined. DESIGN: This was a randomized, double blind, and parallel study. Twenty-four healthy obese males, 19-49 years of age, with body mass index (BMI) > 30 kg/m2 and a waist:hip ratio > 0.95, were treated with MK-677 (25 mg/day; n = 12) or placebo (n = 12) for 8 weeks. RESULTS: MK-677 treatment increased serum leptin levels and leptin/body fat ratio at 2 weeks of treatment (P < 0.05 vs. placebo) but no significant change was observed at 8 weeks. An increase in serum free 3, 5, 3'-triiodothyronine (free T3) was not detected until 8 weeks of MK-677 treatment (P < 0.05 vs. placebo). Peak serum thyroid stimulating hormone (TSH) concentration after MK-677 administration was similar to that after placebo administration at initiation of treatment and at 2 weeks. At 8 weeks of MK-677 treatment, mean peak serum TSH concentration was increased (P < 0.05 vs. placebo) although it remained within the normal range. Serum peak values of luteinizing hormone (LH) and follicle stimulating hormone (FSH) were similar after MK-677 and placebo administration. MK-677 treatment reduced serum total testosterone (P < 0.05 vs. placebo) although total testosterone/sex hormone-binding globulin (SHBG) ratio (an index of free testosterone) was not changed. CONCLUSION: Treatment with the oral GH secretagogue MK-677 transiently increased serum leptin levels and leptin/body fat ratio at 2 weeks of treatment, and increased serum free T3 after 8 weeks. These results indicate that MK-677 treatment is able to affect circulating factors of importance for adipose tissue mass and fuel metabolism. PMID- 10468904 TI - Endothelial dysfunction in hypopituitary adults with growth hormone deficiency. AB - OBJECTIVES: Adult hypopituitarism with growth hormone deficiency (GHD) results in reduced exercise capacity, detrimental changes in body composition and lipid profiles and may be associated with an excess cardiovascular mortality. Endothelial dysfunction is an early event in the pathogenesis of atherosclerosis and predisposes to the deposition of unstable atherosclerotic plaques. We have used a noninvasive method to assess endothelial function in the brachial arteries of a group of treated hypopituitary adults with GHD, and a group of healthy age- and sex-matched controls. PATIENTS: Seventeen hypopituitary adults with GHD (13 male, 4 female) aged 26-54 years were studied. Each patient was receiving standard replacement therapy for all other hormonal deficiencies such that all target hormones were maintained in the normal reference range. All observations obtained were compared with those made in age- and sex-matched control subjects. All study subjects had no identifiable risk factors for endothelial dysfunction. MEASUREMENTS: Using an ultrasound vessel wall tracking system, the diameter of the left brachial artery was measured at rest, in response to reactive hyperaemia (endothelium-dependent dilation) and following sublingual glyceryl trinitrate (GTN) (endothelium-independent vasodilatation). We also measured fasting lipids, insulin, plasma glucose, glycated haemoglobin (HbA1c) and IGF-1, and studied the relationship of these parameters to endothelial function. RESULTS: Flow mediated endothelium-dependent dilatation (FMD), expressed as a percentage change from resting base-line diameter, was significantly impaired in the GHD group (3.70 +/- 2.36% vs. 7.30 +/- 2.42%, P < 0.001). In contrast, GTN-mediated dilatation was similar in both groups. There were no differences in total cholesterol, HDL cholesterol, LDL cholesterol or plasma triglyceride between the groups. Both fasting insulin (27.1 +/- 18.1 vs. 15.89 +/- 6.65 mU/l, P < 0.05) and glycated haemoglobin (HbA1c) levels (5.29 +/- 0.43 vs. 4.91 +/- 0.43%, P < 0.05) were significantly higher in the GHD group. FMD in both groups showed an inverse relationship with total cholesterol (r = -0.58, P < 0.05, GHD and r = -0.55, P < 0.05 controls). However, in the GHD subjects, there was a strong inverse relationship between FMD and LDL-cholesterol (r = -0.81, P < 0.0001). No other relationships were noted between FMD and any other metabolic parameters, or characteristics of GHD. CONCLUSIONS: Endothelial dysfunction is present in GH deficient adults prior to the onset of overt atherosclerotic disease. The similar glucose yet elevated fasting insulin levels imply a state of relative insulin insensitivity. The strong inverse correlation between endothelial dysfunction and LDL-cholesterol suggests a possible aetiological role for LDL-cholesterol in the pathogenesis of any excess cardiovascular risk associated with adult hypopituitarism. PMID- 10468905 TI - Evaluation of adrenal function in patients with hypothalamic and pituitary disorders: comparison of serum cortisol, urinary free cortisol and the human corticotrophin releasing hormone test with the insulin tolerance test. AB - OBJECTIVE: This study aimed to evaluate the performance of screening tests (serum cortisol and 24-h urinary free cortisol) and the human-corticotrophin releasing hormone (h-CRH) test in the assessment of adrenal function in patients with hypothalamic-pituitary disorders. DESIGN: Summary receiver operating characteristics (SROC) curve analysis was applied with the insulin tolerance test (ITT) as reference test. A peak serum cortisol response to ITT > or = 500 nmol/l indicated adrenal sufficiency. The sensitivity at the intersect of the diagonal between sensitivity = 1 and (1-specificity) = 1 with the SROC curve, where sensitivity and specificity are equal, and the corresponding weighted kappa, an estimate of agreement with the ITT, served as parameters of test performance. The diagnostic yield, representing the proportion of tests obviating the need for an ITT, was also calculated. MEASUREMENTS: Serum cortisol at 0800 h (n = 122), at 1600 h (n = 116), 24-h urinary free cortisol (n = 115) and the peak serum cortisol to h-CRH (n = 129) were compared with the peak serum cortisol to ITT. PATIENTS: Eighty patients with hypothalamic-pituitary disorders in whom 75 ITT's were performed pre- and 57 post-operatively. RESULTS: Sensitivity at the intersect and weighted kappa were higher for 0800 h serum cortisol (0.873 and 0.763 respectively) than for 1600 h serum cortisol (0.769 and 0.561) and 24-h urinary free cortisol (0.777 and 0.576). These parameters were 0.868 and 0.756 for the h-CRH test. The diagnostic yield was 63.9% for 0800 h serum cortisol compared to 25.9% for 1600 h serum cortisol (P < 10(-8)), 23.5% for 24-h urinary free cortisol (P < 10(-8)) and 60.5% for the h-CRH test (NS). CONCLUSIONS: Serum cortisol measurement at 0800 h is better than 1600 h and 24-h urinary free cortisol to evaluate adrenal function in this patient category. The diagnostic applicability of the h-CRH test is not superior to 0800 h serum cortisol measurement. PMID- 10468906 TI - Circadian variation of plasma cortisol in prepubertal children with normal stature, short stature and growth hormone deficiency. AB - OBJECTIVES: When studying the relationship between spontaneous secretion of growth hormone (GH) and cortisol in children, most studies show no correlation in mean levels of these two hormones, while others found positive or even strongly negative correlations. These contradictory results could be partly due to the inability to properly compare hormones that are characterized by circadian and ultradian variations in their secretory profiles. We aim here to study possible differences in rhythm characteristics of plasma cortisol with stature and to compare the circadian secretory patients of cortisol and GH. PATIENTS: We analysed data from 135 prepubertal children: (1) 14 GH-deficient children; (2) 36 children with short stature (2-3 SD below their peer group mean); (3) 57 children with very short stature (3-4 SD below their peer group mean); and (4) a reference group of 28 children with normal stature (+/- 2 SD). Subjects were living in a hospital setting on a diurnal waking (07.30-22.30 h), nocturnal resting routine during sampling, consuming the usual hospital diet at fixed times. MEASUREMENTS: Cortisol and GH concentrations were determined by radioimmunoassay in plasma obtained at about 2-3 h intervals during most of the day and at half-hour intervals between 22.00 and 02.00 h. Circadian rhythm characteristics obtained by least-squares estimation were compared between groups divided according to gender and stature with a parameter test. RESULTS: Show a statistically significant circadian rhythm in cortisol secretion for all groups studied (P < 0.001 in all cases). A comparison of circadian parameters indicated similar characteristics between subjects of short, very short and normal stature. Despite a borderline statistically significant difference in rhythm-adjusted mean and amplitude of GH between nondeficient and GH-deficient children, there was no difference in the circadian pattern of cortisol secretion between these two groups. No correlation was found in circadian mean, amplitude, average, standard deviation, standard error, minimum or maximum between GH and cortisol for any of the groups of children. CONCLUSIONS: Any possible relation between GH and cortisol remains unclear. Moreover, GH-deficient children are not necessarily characterized by either hyper- or hypocortisolaemia. PMID- 10468907 TI - Kallmann's syndrome: is it always for life? AB - OBJECTIVE: Kallmann's syndrome (KS) is defined by the association of olfactory deficit with irreversible, congenital gonadotrophin deficiency (IHH). We present evidence for the existence of a variant form of KS, in which endogenous gonadotrophin secretion recovers spontaneously in later life. DESIGN: Longitudinal clinical study. PATIENTS: Five men with anosmia or severe hyposmia, who originally presented in their late teens or early twenties as a result of severe pubertal delay and were thus presumed to have KS. RESULTS: Spontaneous onset of endogenous gonadotrophin secretion, evidenced by progressive normalization of testicular volume and of serum testosterone concentration, occurred in these men over a period of years following the initial diagnosis. CONCLUSIONS: This variant form of Kallman's syndrome is not well recognized and may well be under-diagnosed. Once full virilization has been induced, males with congenital gonadotrophin deficiency whose testes have significantly increased in size should be reassessed, off androgen replacement therapy, to identify those who no longer require treatment. PMID- 10468908 TI - Effect of acute high dose dobutamine administration on serum thyrotrophin (TSH). AB - OBJECTIVE: The interpretation of thyroid function tests in the setting of severe illness is often complicated by concomitant drug administration which may independently produce changes in thyroid hormone concentrations or even secondary hypothyroidism. Although the effects of dopamine on TSH are well established, the effects of dobutamine, another drug commonly used in the setting of severe illness, on TSH are unknown. The aim of the study was to establish the effect(s) of acute high dose dobutamine on serum TSH concentration. DESIGN AND PATIENTS: Thirty subjects undergoing dobutamine stress echocardiogram were compared to twenty controls. Serum TSH was determined between the hours of 0800 and 1000 h at baseline, at maximum dobutamine infusion (20-50 micrograms/kg/min and 15 minutes after stopping dobutamine. MEASUREMENTS: Serum TSH concentration was measured using a third generation chemiluminescent assay. RESULTS: TSH concentration decreased with time in both dobutamine and control subjects and there was an additional statistically significant effect of dobutamine treatment to decrease TSH. TSH concentration remained within the normal range in all subjects who started with normal TSH concentration and remained above normal in the three dobutamine-treated subjects with elevated TSH at baseline. The dobutamine associated decrease in TSH was still present 15 minutes after discontinuing dobutamine. CONCLUSIONS: These results indicate that acute high dose dobutamine lowers TSH by an unknown mechanism. Additional study with prolonged dobutamine infusion is needed to establish the steady state level and physiological consequences of dobutamine-inhibited TSH. PMID- 10468909 TI - The HLA-DQ associations with Graves' disease in Chinese children. AB - OBJECTIVE: Graves' disease has been documented to be associated with different HLA genes in Caucasians and Chinese adults. The incidence of childhood Graves' disease has been reported to be high in Hong Kong Chinese. The aims of this study were to examine the HLA-DQA1 and DQB1 associations with Graves' disease in Chinese children. PATIENTS AND DESIGN: Sixty-seven Chinese children with Graves' disease (59 girls and 8 boys) and 51 racially matched healthy controls were recruited for the study. Genomic DNA was extracted from venous blood samples. HLA DQ typings were determined by sequence-specific oligonucleotide probing of the respective enzymatically amplified gene. Frequencies of HLA-DQ alleles at each locus were compared between patients and controls using the chi 2-test. RESULTS: The frequency of HLA-DQB1.0303 was increased in the combined male and female patient group [53.7%; relative risk (RR) = 4.22; Pc = 0.005] and female patients (50.8%; RR = 3.76; Pc = 0.018) compared with that in the entire control group (21.2%). HLA-DQB1*201 was protective for Graves' disease (10.4%; RR = 0.20; Pc = 0.006). In contrast to studies in Caucasians, DQA1*0501 was not associated with susceptibility for Graves' disease in Chinese children. CONCLUSIONS: This study confirms that DQB1*0303 is a race-specific susceptible allele for Graves' disease in Chinese. Both susceptible and protective HLA-DQ alleles for Graves' disease in Chinese children are different from those in Caucasians. PMID- 10468910 TI - Cardiac function at rest in hypothyroidism evaluated by equilibrium radionuclide angiography. AB - OBJECTIVE: Previous studies have suggested that hypothyroidism affects both systolic and diastolic cardiac function. We have applied equilibrium radionuclide angiography to the study of heart function at rest in hypothyroidism. DESIGN: A prospective study, evaluating cardiac function at rest in primary hypothyroidism. METHODS: Cardiac function was studied by means of equilibrium radionuclide angiography. Screening echo-Doppler examinations were performed on each patient. PATIENTS: Twenty-six consecutive untreated hypothyroid patients without clinical or echocardiographic cardiac disease and 20 healthy matched controls. RESULTS: Between patients and controls, the time to peak emptying rate (161 +/- 6 msec vs. 144 +/- 6 msec, P < 0.05) and the time to peak filling rate (188 +/- 6 msec vs. 170 +/- 5 msec, P < 0.05), were the only discriminatory parameters. In hypothyroid patients, a trend towards a decrease in diastolic cardiac function, expressed as peak filling rate, was observed: 2.6 +/- 0.1 End Diastolic Volume (EDV)/s vs. 3.0 +/- 0.1 EDV/s, P = 0.06. Within the hypothyroid patient group, the time to peak emptying rate was more prolonged in patients with lower free thyroxine levels (R = -0.60, F = 13.5, P < 0.001). Peak filling rate was decreased in patients with lower free thyroxine levels (R = 0.51, F = 8.4, P < 0.01) whereas the time to peak filling rate was more prolonged (R = -0.62, F = 15, P < 0.001). CONCLUSION: In a large group of consecutive patients presenting with primary hypothyroidism, even in the absence of clinical cardiac disease and echocardiographic abnormalities, clear changes in myocardial performance at rest were observed. The most obvious effect of thyroid hormone deficiency on the heart was a lengthening of both systolic and early diastolic time characteristics. Diastolic rather than systolic cardiac function was influenced by hypothyroidism. PMID- 10468911 TI - Prevalence and significance of thyroid autoantibodies in patients with chronic hepatitis C virus infection: a prospective controlled study. AB - OBJECTIVE: To clarify controversies on the prevalence and clinical significance of thyroid autoimmunity in hepatitis C virus (HCV) infection. DESIGN: A prospective controlled and follow-up study. PATIENTS AND MEASUREMENTS: Serum thyroid microsomal antibody (TMA) and thyroid stimulating hormone were assayed and compared in a consecutive, unselected series of 130 patients with chronic HCV infection, 130 sex/age (+/- 2 years)-matched patients with chronic hepatitis B virus (HBV) infection and 260 matched normal controls. RESULTS: The prevalence of thyroid autoantibodies in male patients with chronic HCV was < 2%. The prevalence of TMA (< 1:400) in female patients with chronic HCV infection was significantly higher than that of HBV controls (22.1 vs. 1.6%; P < 0.001), and higher but not significant compared with normal controls (13.5%). However, the trend of increasing prevalence with age in normal controls was not observed in HCV patients. TMA seropositive female HCV patients were not different from seronegative counterparts in age, duration of infection, HLA haplotype, associated autoantibodies and liver histology but had a significantly higher prevalence of genotype 1b/2b mixed infection (P < 0.01) and anti-GOR (P < 0.05). Of the 23 HCV patients seropositive for thyroid autoantibodies, seven had Hashimoto's thyroiditis, two had Graves' disease and three had received subtotal thyroidectomy. During follow-up, four of 15 female patients showed a 14-16-fold increase in TMA titre and one developed hyperthyroidism. Patients with thyroid autoantibodies did not show a propensity to develop thyroid dysfunction during interferon therapy. CONCLUSIONS: These results suggest a weak association between HCV and thyroid autoimmunity in females. As in the ordinary population with thyroid autoantibodies, they should be evaluated for thyroid status and be followed-up if thyroid autoimmunity is evident. However, seropositivity of thyroid autoantibodies is not a contraindication to interferon therapy. PMID- 10468912 TI - Female sex hormone replacement therapy increases serum free 1,25-dihydroxyvitamin D3: a 1-year prospective study. AB - OBJECTIVE: Currently, hormone replacement therapy is applied successfully to reduce post-menopausal bone resorption. However, the exact mechanism by which oestrogen exerts its effect has not yet been fully elucidated. In order to determine whether changes in the biologically active 1,25-dihydroxyvitamin D3 may be of importance in this process, the concentrations of both total and free 1,25 dihydroxyvitamin D3 in serum were assessed. DESIGN: In 36 post-menopausal women the effect of hormone replacement therapy, with a combination of 17 beta oestradiol and norethisterone acetate, on the serum levels of total and free 1,25 dihydroxyvitamin D3 was studied after 0, 3, 6 and 12 cycles. MEASUREMENTS: The total concentration of 1,25-dihydroxyvitamin D3 in serum was assessed using a radioreceptor assay after diethylether extraction of the samples followed by paper chromatography. The free fraction of 1,25-dihydroxyvitamin D3 was measured using symmetric dialysis. The free 1,25 dihydroxyvitamin D3 concentration was calculated by multiplying the total concentration by the free fraction. RESULTS: During therapy, mean serum total 1,25-dihydroxyvitamin D3 concentrations (+/- SD) were 106.4 pmol/l (+/- 27.5), 155.0 pmol/l (+/- 49.5), 176.7 pmol/l (+/- 70.0) and 161.1 pmol/l (+/- 55.3) at 0, 3, 6 and 12 cycles, respectively. Serum free 1,25-dihydroxyvitamin D3 concentrations were 68 fmol/l (+/- 22), 107 fmol/l (+/- 35), 120 fmol/l (+/- 43) and 108 fmol/l (+/- 37), respectively. Baseline values of both total and free 1,25-dihydroxyvitamin D3 were significantly lower than those during therapy at all time (P < or = 0.001). CONCLUSION: Both the serum total 1,25-dihydroxyvitamin D3 and the serum free 1,25-dihydroxyvitamin D3 concentrations are increased during combined 17 beta-oestradiol and norethisterone acetate therapy for a year. Assuming that the free concentration 1,25-dihydroxyvitamin D3 reflects the biologically active fraction, this rise may in part explain the preventive effect of hormone replacement therapy on osteoporosis. PMID- 10468913 TI - The natural history of the metabolic syndrome in young women with the polycystic ovary syndrome and the effect of long-term oestrogen-progestagen treatment. AB - OBJECTIVE: Little is known about the natural history of polycystic ovary syndrome (PCOS), although preliminary data indicate that affected women are more susceptible than the general population to diabetes and cardiovascular diseases at post-menopausal ages. The aim of this study was to follow-up all main features of the metabolic syndrome in a group of young women with PCOS and to investigate the long-term effects on metabolism and body composition of oestrogen-progestagen (OP) compounds, which are frequently used in these women to treat hyperandrogenism and related clinical features. DESIGN: Long-term follow-up study. SUBJECTS AND METHODS: Thirty-seven women with PCOS were re-evaluated 10.3 +/- 0.8 years (range 6-18 years) after their first assessments (age: before 19.8 +/- 4.9 years; after 29.9 +/- 4.4 years). When first examined, women were instructed to follow a hypocaloric diet if they were obese plus OP, if they agreed to such treatment. Main anthropometric parameters, basal sex hormones and lipids, fasting and glucose-stimulated glucose and insulin levels and several clinical data were recorded before and after follow-up. RESULTS: In the whole group of women with PCOS we found no changes in body weight and fat mass, whereas both the waist-to-hip ratio and the waist-to-thigh ratio were significantly reduced. No significant changes occurred in mean fasting and glucose-stimulated glucose and insulin concentrations, whereas a significant increase in high density lipoprotein-cholesterol was found. No significant changes occurred in testosterone levels. During the follow-up period 16 women took OP for an average of 97 +/- 18 months (range 12-180 months) (OP-users) whereas 21 women never took OP (non-OP-users). All OP-users were still taking OP when re-evaluated at the follow-up examination. With respect to baseline values, body mass index was higher in non-OP-users than in their counterparts. Waist circumference (P < 0.025), the waist-to-hip (P < 0.05) and the waist-to-thigh (P < 0.01) ratios decreased significantly only in the OP-users. In addition, percentage changes in waist circumference (P < 0.05) and waist-to-hip ratio (P < 0.05) during the follow-up period were significantly different between the groups. Glucose tolerance (as area under the curve (AUC)) improved (P < 0.05) in OP-users but not in non-OP-users. Moreover, compared to baseline values, basal insulin levels were significantly (P < 0.01) reduced in OP-users but not in non-OP-users. On the contrary, no significant change was found in insulinAUC in the former, whereas it significantly increased (P < 0.05) in the latter. Accordingly, fasting C-peptide decreased (P < 0.05) in OP-users, whereas both fasting (P < 0.01) and stimulated (P < 0.01) C-peptide significantly increased in non-OP-users. Changes in fasting or stimulated insulin and C-peptide in non-OP-users were not associated with parallel changes in testosterone levels. Total cholesterol and triglycerides did not change in either group, but HDL-cholesterol increased (P < 0.05) only in OP users. Sex hormone-binding globulin concentrations increased significantly (P < 0.01) in OP-users, without any significant change in non-OP-users. Testosterone concentrations did not change significantly in either group, but the testosterone: SHBG ratio significantly decreased in OP-users (P < 0.05) but not in the non-OP-users. Among the clinical features, acanthosis nigricans significantly (P < 0.01) worsened in non-OP-users but not in the OP-users, without any significant change in the hirsutism and acne scores. Pregnancy rates during the follow-up were similar in both groups. CONCLUSIONS: These data indicate that hyperinsulinaemia and insulin resistance tended to worsen spontaneously in women with PCOS, without any worsening of the hyperandrogenism. Long-term oestrogen-progestagen treatment countered this tendency, probably because it improved the pattern of body fat distribution, by reducing abdominal fat depots. PMID- 10468914 TI - Prevalence of Ras mutations in thyroid neoplasia. AB - OBJECTIVE: Mutations at codons 12, 13 or 61 of ras which result in constitutive activation occur frequently in human malignancies. There have been varied reports on their prevalence and hence their likely significance in the pathogenesis of primary thyroid neoplasia. To address this, we have examined a large series of benign and malignant thyroid tumours for ras mutations. DESIGN: Genomic DNA was analysed for the presence of mutations at codons 12, 13 and 61 of H-ras, K-ras and N-ras by allele-specific oligonucleotide hybridization. Direct DNA sequencing was used to confirm the mutations. PATIENTS: A total of 90 samples with benign (66) and malignant (24) thyroid disease were investigated. RESULTS: A total of 14/90 (15.5%) samples had a ras mutation. All mutations were at codon 61 of either N-ras or K-ras. The positive cases were 1/25 (4%) nodular goitre, 7/38 (18%) follicular adenoma, 4/9 (44%) follicular carcinoma, 1/1 anaplastic carcinoma, 1/1 follicular variant of papillary carcinoma, and 1 metastatic follicular carcinoma in which the primary tumour had the same mutation. CONCLUSIONS: Our data demonstrate a relatively low overall prevalence of ras mutations in thyroid neoplasia, with a predominance in follicular neoplasms. Their presence in follicular adenomas suggests that they may have an early aetiological role in the development of thyroid neoplasia. PMID- 10468915 TI - An adult patient with severe hypercalcaemia and hypocalciuria due to a novel homozygous inactivating mutation of calcium-sensing receptor. AB - Inactivating mutations in the calcium-sensing receptor (CaSR) cause familial hypocalciuric hypercalcaemia (FHH) and neonatal severe hyperparathyroidism (NSHPT). Earlier investigations showed patients with FHH are heterozygous, and NSHPT are homozygous for inactivating mutations. However, one adult patient with severe hypercalcaemia and hypocalciuria has been reported to have a homozygous inactivating mutation in CaSR (Pro39Ala). This suggested that mutant CaSR in this patient had some residual activity and hypercalcaemia was not so severe as to be fatal. However, the function of this mutant CaSR was not evaluated. In the present study, we describe a novel homozygous mutation in an adult patient with severe hypercalcaemia and hypocalciuria, and evaluate the function of the mutant CaSRs. The DNA sequence of CaSR gene was determined by direct sequencing of the polymerase chain reaction product. The function of mutant CaSR was analysed by creating mutant cDNAs by in vitro mutagenesis, transfection of mutant cDNAs into HEK293 cells and measuring intracellular ionized Ca in response to changes in extracellular Ca. A 26-year-old Japanese woman showed marked hypercalcaemia with an elevated parathyroid hormone (PTH) level. Her consanguineous parents had asymptomatic hypercalcaemia with relative hypocalciuria. The proband had a homozygous mutation at codon 27 of CaSR gene (CAA-->CGA, Gln27Arg). Her parents were heterozygous for this mutation. EC50 for Ca of this mutant CaSR (GIn27Arg) was 4.9 mM. EC50 of another mutant CaSR (Pro39Ala) whose homozygous mutation was discovered in an adult patient was 4.4 mM. These EC50s were significantly higher than that of wild-type CaSR (3.7} 0.1 mM), but were the lowest among the reported EC50s for inactivating mutations of CaSR. These results indicate that serum Ca and PTH levels are determined by residual function of mutant CaSR in patients with homozygous mutation in CaSR, and that patients having homozygous mutant CaSRs with mild dysfunction do not suffer from fatal hypercalcaemia in infancy and can survive into adulthood. PMID- 10468916 TI - Medullary thyroid carcinoma in Graves' disease. PMID- 10468917 TI - Glucocorticoid therapy for nonislet cell tumour hypoglycaemia (NICTH) PMID- 10468918 TI - Can GHRH or GH secretagogues re-initiate pituitary GH pulsatility? PMID- 10468919 TI - How many surgeons to operate on acromegalic patients? PMID- 10468920 TI - Outcome of transphenoidal surgery for acromegaly and its relationship to surgical experience. AB - BACKGROUND: A number of surgical series have been reported on the treatment of acromegaly and their results vary widely. The acceptable definition of remission has changed in recent years and it is known, though in a small series, that growth hormone levels of > 5 mU/l are still associated with an increased mortality from the condition. We have analysed data at this centre and examined the outcome of transphenoidal surgery for acromegaly, compared our results with recently published series from other centres and also assessed factors which might effect outcome including whether there is any demonstrable effect of the experience of the surgeon on outcome. PATIENTS AND METHODS: We have analysed data from all of our 139 patients in whom follow up data are available who have undergone initial transphenoidal surgery for acromegaly by one surgeon at this centre, between 1974 and 1995. Follow up was for a median of 5 years (range 1 month to 17 years). RESULTS: 67% of patients achieved the criterion for remission (mean GH < 5 mU/l). Success was related to tumour size and preoperative growth hormone values. Thus 91% of patients with microadenomas were in remission postoperatively compared to 46% of patients with macroadenomas. Analysis of the results according to the year of operation showed an improvement in success rates with time. More than 15 years ago, the success rate according to the growth hormone criteria set was 48% and the failure rate 52%. In contrast in the last 5 years analysed, the overall success rate was 74% with a failure rate of 26% (P < 0.04). The success rate for microadenomas was 50% initially, then remained 100%. The case mix was analysed and no change was found. We have also demonstrated an improvement in pituitary function (including normalization of preoperative elevated prolactin) with time so that pre 1981 16% of patients' pituitary function improved perioperatively but 10 years later this figure had risen to 34% (P < 0.03). There was no change over time in the development of pituitary hypofunction, complication rate or recurrence rate. CONCLUSION: Surgical treatment is a safe and effective treatment for acromegaly and remains the first choice of treatment for most acromegalic patients. The results of this centre compare favourably with series from other centres. We have demonstrated improved results, both in terms of post operative growth hormone values and pituitary function tests with time and increasing neurosurgical experience. We conclude that outcome for the surgical treatment for acromegaly is best achieved with one surgeon specialising in pituitary surgery. Improved operative outcome thus achieved has major cost implications and avoids the necessity for consideration of postoperative radiotherapy and the use of expensive growth hormone suppressing drugs in the postoperative period. PMID- 10468921 TI - Insulin regulation of leptin synthesis and secretion in humans: the model of myotonic dystrophy. AB - OBJECTIVE: Myotonic dystrophy (MyD) is a systemic disorder in which insulin resistance is well recognized. In the present study we have characterized plasma leptin levels in patients with MyD and in age, sex and body mass index (BMI) matched controls and assessed the influence of leptin on the clinical manifestations of MyD. DESIGN AND PATIENTS: Body composition, plasma leptin, fasting and post-oral glucose tolerance test insulin, IGF-I and IGFBP3 were studied in 34 MyD patients and 33 controls. MEASUREMENTS: Body composition was measured using a bioelectrical impedance analyzer, and circulating levels of insulin, leptin, IGF-I, IGFBP3 were measured by IRMA or RIA. Insulin sensitivity was modelled according to a homeostasis model assessment (HOMA) computer-solved model. RESULTS: Percentage body fat was higher in patients than in controls (25.6 +/- 2.28% vs 18.8 +/- 1.53%, P = 0.013). Insulin levels, both fasting and after oral glucose were higher in patients than in controls, and insulin sensitivity was lower in patients than in controls. Serum leptin was higher in patients than in controls (20.98 +/- 3.11 micrograms/l vs 10.4 +/- 1.31 micrograms/l, P = 0.004), and higher in women than in men, both in patients and in controls. In patients, leptin levels were correlated with age, BMI, fasting insulin, insulin area under curve and lower insulin sensitivity, whereas leptin levels were not correlated with body fat or other parameters of body composition. In controls, leptin levels were correlated with BMI and body fat. The results were evaluated using logistic regression models for each of the 2 populations. In the model of MyD, insulin resistance and age correctly identified higher leptin levels in relation to controls out of 87.88% of patients, and in the model of controls male sex with a negative correlation and BMI correctly identified their leptin levels out of 84.33% cases. CONCLUSIONS: These findings show that MyD provides a different model of leptin regulation in humans, and suggest that in MyD patients there are correlations between leptin and insulin resistance and age, irrespective of body fat. In contrast, leptin levels in controls, correlate with sex and BMI. The data on leptin in this population of patients can not be related aetiologically to the muscle disease itself. PMID- 10468922 TI - A trial of growth hormone therapy in well-controlled hypophosphataemic rickets. AB - OBJECTIVE: Conventional therapy of hypophosphataemic rickets (HR) with oral phosphate and calcitriol does not always result in normal linear growth. Recombinant human growth hormone (rhGH) offers theoretical advantages as an adjunctive therapy. We aimed to determine the effects of adjunctive rhGH therapy in children with well-controlled HR. PATIENTS: In this report, 5 prepubertal children (aged 3.5-10.9 years) with well-controlled HR on conventional therapy were given adjunctive standard dose rhGH therapy for one year. DESIGN AND MEASUREMENTS: Height, growth velocity, metabolic markers of calcium and phosphate metabolism, body composition, bone mineral density, wrist and knee X-rays, and renal sonography were assessed at regular intervals. Height and growth velocities were also calculated 12 months after ceasing rhGH therapy. RESULTS: After 12 months therapy with rhGH, no significant biochemical or radiological benefits were observed. A significant increase in height SD score was observed (P = 0.023), but this was not associated with any increase in the growth velocity SD score and appears to have been due to catch-up growth caused by conventional therapy alone. When rhGH therapy was ceased, no significant decreases in mean height SD or growth velocity SD scores were observed. CONCLUSIONS: In well controlled hypophosphataemic rickets patients receiving conventional therapy, adjunctive therapy with standard dose rhGH offers no benefits in linear growth or rachitic disease markers. PMID- 10468923 TI - Parathyroid hormone gene polymorphisms in primary hyperparathyroidism. AB - OBJECTIVE: Genetic contributions to bone mineral density (BMD) and bone turnover are well known. In the present study, we analysed the relationship between restriction fragment length polymorphisms of the PTH gene and the development of primary hyperparathyroidism (pHPT) as well as its severity. PATIENTS: Seventy nine pHPT patients and 104 age-matched healthy controls were analysed. DESIGN AND MEASUREMENTS: PTH genotypes were determined by polymerase chain reaction and BstB I or Dra II restriction fragment length polymorphisms. The presence and absence of BstB I or Dra II restriction sites of the PTH gene were indicated by B and b or D and d, respectively. BMD levels at the lumbar spine and at the radius were measured in all subjects. Serum levels of calcium, phosphorus, alkaline phosphatase and intact PTH were measured in pHPT patients. RESULTS: There were no differences in the frequencies of these PTH genotypes between pHPT patients and controls. In control subjects, lumbar BMD was significantly higher in BB genotype than in Bb/bb genotypes. In pHPT patients, there was no difference of BMD between BB and Bb/bb genotypes. In pHPT patients, serum calcium level was significantly higher in those with the BB genotype than Bb/bb genotypes. On the other hand, there was no association between Dra II polymorphism and BMD in both controls and pHPT patients, but serum intact PTH level was significantly higher in DD genotype than Dd/dd genotype in pHPT patients. Moreover, serum levels of ALP and intact PTH were significantly higher in the PTH BBDD haplotype, compared to those in haplotypes other than BBDD. CONCLUSIONS: The present study suggests that the BstB I polymorphism of PTH gene is closely related to bone mineral density and that PTH gene polymorphisms do not seem to affect the development of primary hyperparathyroidism but may relate to the severity of primary hyperparathyroidism. PMID- 10468924 TI - Familial non-medullary thyroid carcinoma: pathology review in 27 affected cases from 13 French families. AB - BACKGROUND AND OBJECTIVES: When familial non-medullary thyroid cancer (FNMTC) develops with no obvious associated pathogenetic factor, an inherited predisposition may underlie the process. The present study was conducted because detailed pathological findings are lacking in most series of FNMTC. PATIENTS AND METHODS: Thirteen families comprising 27 cases of FNMTC were included (1.8% of differentiated thyroid carcinoma). The family relationship (20 F, 7 M; age 46 +/- 16 years; mean +/- SD) was 'siblings' in eight families, 'parent and child' in four and 'aunt and niece' in one. Careful pathological review of the thyroid tumours (papillary/follicular: 25/2, size: 16 +/- 11 mm) was performed. RESULTS: Initial staging according to extension was as follows: grade I (n = 16), II (n = 2), III (n = 6), IV (n = 3). Fourteen tumours were papillary microcarcinomas (size: 8 +/- 2 mm). No tumour phenotype that may be considered specific for FNMTC was found when considering either age, pathological findings or tumour aggressiveness. Although rare events were found in both relatives of some families suggesting a putative 'familial' phenotype of FNMTC, this may be fortuitous. CONCLUSION: Micro familial non-medullary thyroid cancers are more common than previously reported and further studies are required to be able to distinguish this subgroup from sporadic papillary microcarcinomas. The careful pathological review of the familial non-medullary thyroid cancer in this study does not seem to point to a distinct subgroup of familial differentiated thyroid carcinoma although the data are intriguing. Genetic studies are now required to investigate this issue. PMID- 10468925 TI - Relationships between dehydroepiandrosterone-sulphate and anthropometric, metabolic and hormonal variables in a large cohort of obese women. AB - OBJECTIVE: The aim of the present study was to measure dehydroepiandrosterone sulphate (DHEA-S) levels in obesity and assess the relationships between DHEA-S and anthropometric, metabolic and hormonal variables. SUBJECTS AND METHODS: We evaluated the serum DHEA-S levels in 217 obese but otherwise normal female subjects (age (mean +/- SEM): 39.4 +/- 0.9, range 18-67 years, body mass index (BMI) = 36.1 +/- 0.4, range 27.1-57.1 kg/m2). RESULTS: DHEA-S levels showed an age-dependent fall similar to that observed in normal women (n = 156, age 46.2 +/ 1.2, range 22-69 years, BMI < 25 kg/m2). Adjusting for age, obese women had mean DHEA-S levels higher than the control group (P < 0.02). In obese patients, DHEA-S levels were directly associated with serum testosterone, androstendione, IGF-I, fT3 levels and 24 h-urinary cortisol. On the other hand, DHEA-S levels were negatively associated with age, total cholesterol, triglycerides levels and systolic blood pressure. No correlation was found with BMI, waist:hip ratio, basal and post-OGTT insulin and glucose, free fatty acids, GH, PRL, fT4, TSH, SHBG levels or diastolic blood pressure. Multiple regression analysis indicated that in obese women, DHEA-S levels were associated negatively to age and positively to testosterone, androstendione and IGF-I levels and daily urinary cortisol. In a subgroup of 20 obese women, DHEA-S levels significantly (P < 0.001) fell after OGTT without any correlation with the insulin response. CONCLUSIONS: The present results show that dehydroepiandrosterone-sulphate levels are not reduced in obesity, being slightly increased, particularly in young adulthood. Dehydroepiandrosterone-sulphate levels are positively and independently associated with androgen, 24-h urinary cortisol and IGF-I levels but do not seem associated with insulin levels or cardiovascular risk indices. PMID- 10468926 TI - Circulating levels of human insulin-like growth factor binding protein-6 (IGFBP 6) in health and disease as determined by radioimmunoassay. AB - OBJECTIVE: Insulin-like growth factor binding protein-6 (IGFBP-6) is a relatively unknown member of a family of six specific structurally related IGF binding proteins which are involved in the modulation of the biological effects of the IGFs. A distinctive property of IGFBP-6 is its preferential affinity for IGF-II relative to IGF-I. In order to obtain more insight into the clinical significance and regulation of circulating levels of IGFBP-6 we developed a specific radioimmunoassay (RIA) for this protein. DESIGN AND PATIENTS: Selected human biological fluids and plasma from 847 normal subjects were analysed. In addition, plasma samples from patients with different disorders (i.e. GH-deficiency, acromegaly, cancer, corticosteroid-treated children suffering from different kinds of severe illness and chronic renal failure) were investigated. MEASUREMENTS: The IGFBP-6 assay is competitive, utilizing a rabbit polyclonal antibody raised against a synthetic peptide comprising amino acids 90-118 of the hIGFBP-6 sequence and an additional tyrosine residue. It is calibrated against recombinant human (rh)IGFBP-6. The 125I tracer is prepared by iodination of the synthetic peptide. There is no significant cross-reactivity with other IGFBPs and no interference with the IGFs. RESULTS: Extensive normative range values for IGFBP-6 were determined using 847 plasma samples from normal males and females, ranging from 0 to 75 years of age. IGFBP-6 levels increased gradually (about two fold) with age. In childhood the plasma levels of IGFBP-6 in females tended to be slightly higher than those for males. For the adult population the reverse was observed. Overall, the mean +/- SD value for males was higher than that for females (149 +/- 57 vs. 139 +/- 45 micrograms/l, P < 0.004). GH status did not appear to influence IGFBP-6 level since normal levels were found for both untreated acromegalic patients and GH-deficient subjects. GH treatment of the latter group of patients did not alter IGFBP-6 in plasma. Pharmacological doses of glucocorticosteroids affected circulating IGFBP-6 levels only slightly. IGFBP 6 levels in plasma samples derived both from children with acute lymphoblastic leukaemia and from patients with various types of solid neoplasms were generally within the normal range. In contrast, plasma samples from four of six patients with non-islet cell tumour induced hypoglycaemia (NICTH) showed elevated concentrations of IGFBP-6 (SDS > 2.9). An excess of IGFBP-6 was also found in plasma of both dialysed and non-dialysed prepubertal growth retarded children with chronic renal failure (CRF) (mean SDS: 23.0 and 9.3, respectively). IGFBP-6 levels were inversely correlated with glomerular filtration rate. In a group of CRF patients who underwent renal transplantation circulating IGFBP-6 levels were markedly lower (mean SDS: 4.6). The presence of IGFBP-6 could also be demonstrated in several other human biological fluids. Low amounts were detected in saliva (3-12 micrograms/l) and breast milk (6-45 micrograms/l) while the levels in amniotic fluid and follicular fluid were comparable with those determined in normal plasma. The IGFBP-6 content of cerebrospinal fluid (CSF) ranged between 25 and 87 micrograms/l, which is rather high in relation to the relatively low concentration of total protein in this body fluid. CONCLUSIONS: Measurements of IGFBP-6 have been shown so far to be of relatively minor clinical relevance. The exceptions are chronic renal failure patients and subjects with large tumours and non-islet cell tumour induced hypoglycaemia who may exhibit elevated circulating levels of this IGFBP. The physiological significance of this observation remains to be elucidated. The possibility of quantifying IGFBP-6 by specific RIA will facilitate further in vitro and in vivo studies of its regulation and function in man. PMID- 10468927 TI - Relationship between serum and urinary insulin-like growth factor-I through childhood and adolescence: their use in the assessment of disordered growth. AB - OBJECTIVE: Serum insulin-like growth factor-I (sIGF-I) measurement as an index of growth hormone status has become a common test in the investigation of disordered growth. IGF-I may also be measured in the urine. The aims of this study were to investigate the correlation between serum and urinary IGF-I in normal children and compare their use in the evaluation of growth disorders. DESIGN: Normal ranges for serum and urinary IGF-I were devised from a cross-sectional study of normal schoolchildren. These were then used to assess the sensitivity and specificity of serum and urinary IGF-I in the diagnosis of childhood GH deficiency. PATIENTS: A cohort of 333 (M = 156, F = 177) healthy schoolchildren aged 5-19 years were recruited and data previously collected from 22 growth hormone deficient (GHD) and 47 short normal (SN) children were compared with those of the normal children. MEASUREMENTS: Height, weight and pubertal status were assessed in all children. Serum IGF-I (sIGF-I) (n = 305) and total amount of urinary IGF-I excreted overnight (TuIGF-I) (n = 205) were measured by RIA using excess IGF-II to block the interference of IGFBPs. RESULTS: Serum IGF-I was loge transformed and overall levels (geometric mean +/- 1 tolerance factor) were higher in females than males (F: 569 (329, 985) micrograms/l; M: 398 (227, 696) micrograms/l). LogeIGF-I correlated with age (F: r = +0.76, P < 0.001, M: r = +0.71, P < 0.001) and was significantly affected by both sex and Tanner stage of puberty (TS) (both P < 0.001). The distribution of TuIGF-I was normalized by performing a square root transformation (square root of TuIGF-I). square root of TuIGF-I was correlated with age (F: r = +0.36, P < 0.001; M: r = +0.5, P < 0.001) and was significantly affected by TS (P < 0.001). In both sexes there was a highly significant correlation between logeIGF-I and square root of TuIGF-I (F: r = +0.39, P < 0.001; M: r = +0.41, P < 0.001). Using the third centile of our normal ranges as a cut off to identify GHD, sIGF-I had a sensitivity of 82% and specificity of 62%, whereas TuIGF-I had a sensitivity of 18% and specificity of 79%. CONCLUSIONS: This study demonstrates that although urinary IGF-I has no place in the diagnosis of growth disorders, in normal children there is a highly significant relationship between serum and urinary IGF-I with levels of each changing in a similar manner through childhood and adolescence. Thus, TuIGF-I could be used as a valid surrogate for sIGF-I in the physiological assessment of the relationship between IGF-I status and the normal growth process. PMID- 10468928 TI - Acidic isoforms of chorionic gonadotrophin in European and Samoan women are associated with hyperemesis gravidarum and may be thyrotrophic. AB - OBJECTIVE: There is conflicting evidence concerning the role of human chorionic gonadotrophin (hCG) in the aetiology of hyperemesis gravidarum (HG); particular isoforms of hCG may be the critical factor. Ethnic differences in HG prevalence and putative thyrotrophic effects of hCG may also relate to differences in hCG isoform profiles. To address these issues we examined the relationship of hCG isoforms to HG and thyroid function tests in two groups of women from ethnic backgrounds with significantly different HG prevalence rates. PATIENTS AND DESIGN: We enrolled 10 European and 10 Samoan women with HG and an equally sized non-hyperemetic, gestational stage matched control group. MEASUREMENTS: We administered a questionnaire, generated serum hCG charge-isoform profiles by chromatofocusing and measured the serum concentrations of total hCG, oestradiol (E2), thyrotrophin (TSH) and free thyroxine (FT4). RESULTS: The mean serum total hCG levels were highest in the Samoan hyperemetics (176,268 IU/l), and overall higher in hyperemetics compared with controls (159,770 IU/l vs. 86,420 IU/l, P < 0.001). When compared with controls, hyperemetics displayed increased hCG concentrations in the more acidic half (pH < 4) of the chromatofocusing pH range (89,843 IU/l vs. 41,146 IU/l, P < 0.003). Serum E2 levels did not differ between the four groups, but correlated with the hCG concentration between pH 5.2 and 4.01. Mean serum TSH levels were significantly lower in hyperemetics than in controls (0.33 mIU/l vs. 1.19 mIU/l, P < 0.001) and correlated with the hCG concentration between pH 4.6 and 2.8, while serum FT4 correlated with the hCG concentration below pH 4.0. CONCLUSIONS: Acidic isoforms of hCG may play a role in the aetiology of HG and gestational thyrotoxicosis. Minor ethnic differences in hCG isoform profiles were observed, but the relationship of acidic hCG isoforms to HG and serum thyroid hormone levels was largely independent of the patients' ethnicity. The mechanisms by which acidic isoforms might provoke nausea remain uncertain, but do not seem to involve E2, while the longer half-life of acidic hCG isoforms may result in increased in vivo TSH receptor cross-talk with resultant thyrotrophic effects. PMID- 10468929 TI - Long-term substitution therapy of hypogonadal men with transscrotal testosterone over 7-10 years. AB - OBJECTIVE: Testosterone (T) substitution of hypogonadal men by conventional intramuscular injection of T esters is not considered optimal because it induces unphysiologically fluctuating serum T levels. In contrast, scrotal T patches produce normal serum (T) levels mimicking diurnal variations. In order to assess the quality of this new form of T substitution we followed hypogonadal men treated by transdermal T up to 10 years. PATIENTS: Eleven men aged 35.9 +/- 9.8 years (mean +/- SD) at the beginning of the study were treated with transscrotal T patches (Testoderm) because of primary (n = 4) or secondary (n = 7) hypogonadism. Clinical examinations were performed every 3 months during the first 5 years and every 6 months thereafter. All 11 patients were seen for 7 years and some for longer periods: eight for 8 years, six for 9 years and four for 10 years. MEASUREMENTS AND RESULTS: With daily application of one patch T levels rose from 5.3 +/- 1.3 nmol/l (mean +/- SE) to 16.7 +/- 2.6 nmol/l at month 3 and remained in the normal range throughout treatment. Serum 5 alpha dihydrotestosterone (DHT) rose from 1.3 +/- 0.4 nmol/l to 3.9 +/- 1.4 nmol/l and oestradiol from 52.3 +/- 9.3 to 71.3 +/- 9.6 pmol/l and remained stable without significant variations throughout the observation period. Patients reported absence of local side-effects except for occasional itching. No relevant changes occurred in clinical chemistry, including total cholesterol levels and triglycerides. Haemoglobin and erythrocyte counts remained normal. Bone density measured by QCT increased slightly from 113.6 +/- 5.4 to 129.7 +/- 9.3 mg/cm3 during the observation period (P = 0.028). In the nine patients aged < 50 years prostate volumes showed a small but insignificant increase from 16.8 +/- 1.5 to 18.8 +/- 2.1 ml during transscrotal T therapy. In the two older patients prostate volume remained constant or decreased slightly during T therapy after an initial increase in the previously untreated patient. Prostate specific antigen levels were constantly low in all patients. CONCLUSION: Transscrotal testosterone patches are well-accepted and safe in long-term testosterone substitution therapy for male hypogonadism. PMID- 10468930 TI - Expression of the androgen metabolizing enzyme UGT2B15 in adipose tissue and relative expression measurement using a competitive RT-PCR method. AB - OBJECTIVES: We have demonstrated previously that obesity in men was significantly associated with low plasma testosterone levels and higher concentrations of the androgen metabolite androstane-3 alpha, 17 beta-diol glucuronide, suggesting that androgen metabolism and elimination is increased in this condition. The objective of the present study was to investigate whether adipose tissue was a site of expression of the androgen metabolizing enzymes UDP-glucuronosyltransferases (UGT) 2B15 and 2B17. DESIGN AND PATIENTS: Subcutaneous and visceral adipose tissue was obtained from male patients subjected to various abdominal surgeries. MEASUREMENTS AND RESULTS: By performing reverse transcriptase-PCR (RT-PCR) amplification of mRNA extracted from adipose tissue samples, UGT2B15 transcript was detected in both subcutaneous and omental adipose tissue while UGT2B17 transcript expression was very low, or undetectable. A quantitative, competitive RT-PCR method was established and used to quantify UGT2B15 messenger RNA. The level of UGT2B15 expression was also measured in other human tissues. Although the major sites of expression were the liver and the lung, expression in adipose tissue was similar to levels found in the prostate, testis and mammary gland. CONCLUSIONS: These results demonstrate for the first time that both subcutaneous and visceral adipose tissue express androgen metabolizing enzyme UGT2B15 mRNA and further support the role of adipose tissue as a site of steroid metabolism. PMID- 10468931 TI - Differentiated thyroid cancer: a retrospective analysis of 832 cases from Greece. AB - OBJECTIVE: To analyse the initial manifestations, pathological findings, therapy, outcome and prognostic factors in patients with papillary and follicular carcinoma. PATIENTS: 832 patients with well differentiated thyroid carcinoma managed in our department during a period of 30 years (1965-1995). Follow-up data were available for 609 patients for a mean of 5.5 years (range 1-38 years), the remainder having been lost to follow-up. RESULTS: The patients were 677 (81%) with papillary and 155 with follicular carcinoma. They were predominantly female (75%), presenting mainly with a single nodule (53%), while at the time of diagnosis 72% had intrathyroidal carcinomas (class I), 17% had nodal metastases (class II), 7% soft tissue invasion (class III) and 4% distant metastases (class IV). Fifty-five percent of the patients had a complete thyroidectomy (36% had a near total or total thyroidectomy and 19% near total or total thyroidectomy plus block dissection), 2.6% received external radiotherapy and 94% had radioactive iodine as part of the treatment of the original disease. Kaplan-Meier survival analysis was used to calculate both cancer related mortality and disease free survival in the patients followed-up. Although mortality (21 cancer-related deaths) was slightly higher for follicular than papillary carcinoma (10% vs. 5% and 16% vs. 10% in 10- and 15-year survival, respectively) the difference was not statistically significant. Extent of disease at diagnosis, male sex, tumour size and age > 60 years affected probability of cancer death. Cox's proportional hazard regression analysis for disease free survival showed that adverse independent prognostic factors were, for papillary carcinoma, male sex, class II or higher, tumour size > 1 cm and age > 60 years, while for follicular, class III or higher, size > 4 cm and age > 40 years. CONCLUSIONS: We conclude that there is a higher prevalence of follicular carcinoma in our country probably due to a moderate degree of iodine deficiency still existing in Greece. Age and extent of disease at diagnosis were important prognostic factors affecting morbidity and mortality, whereas sex, tumour features and histological type were of minor importance. All these prognostic factors and their relative importance should be taken in consideration in the management of this disease. PMID- 10468932 TI - Disturbances of menstruation in hypothyroidism. AB - OBJECTIVES: Most thyroid textbooks describe hypothyroidism resulting in changes in the length of the menstrual cycle and in the amount of bleeding. Our experience at a busy thyroid clinic had given a picture which differs from that presented in the literature. The aim of this study was to determine the frequency and type of menstrual irregularities in premenopausal hypothyroid patients. DESIGN AND PATIENTS: We investigated the menstrual history, starting 6 months before the discovery of the disease, in 171 female premenopausal hypothyroid patients and 214 normal controls, with similar age and body mass index (BMI). MEASUREMENTS: TSH, TT4, TT3 and thyroid antibodies were measured by radioimmunoassay, while BMI was calculated from the ratio of body weight in kg to height in m2. RESULTS: Of the 171 hypothyroid patients, 131 (76.6%) had regular cycles and 40 (23.4%) irregular periods. Oligomenorrhoea and menorrhagia were the most common features in the latter group. No difference in BMI was found between the patients with or without menstrual abnormalities, or between patients and controls. TSH, TT4 and TT3 levels did not differ between patients with normal and abnormal menstruation. Forty-six (26.9%) out of 171 patients had subclinical hypothyroidism (group A), 42 (24.6%) mild hypothyroidism (group B) and 83 (48.5%) severe hypothyroidism (group C). Thirty-seven from group A (80.4%), 32 from group B (76.2%) and 62 from group C (74.7%) had normal periods. These differences were not statistically significant. Eighty-eight (88.9%) out of 99 patients in whom thyroid antibodies were measured had positive titres. Nineteen (21.6%) of the patients with positive thyroid antibodies had menstrual irregularities. This percentage did not differ from that found in the whole group of patients (23.4%). Out of 214 normal controls, 196 (91.6%) had normal menstruation and 19 (8.4%) irregular cycles. The latter group included mainly women with oligomenorrhoea. CONCLUSIONS: These data demonstrate that hypothyroidism in women is less frequently associated with menstrual disturbance than was previously described. Also, menstrual irregularities tend to be more frequent in sever hypothyroidism in comparison with mild cases, although this finding was not statistically significant. Oligomenorrhoea and menorrhagia are the most common menstrual disturbances. PMID- 10468933 TI - Adrenal and gonadal contributions to urinary excretion and plasma concentration of epitestosterone in men--effect of adrenal stimulation and implications for detection of testosterone abuse. AB - OBJECTIVE: The ratio of urinary testosterone (T) to epitestosterone (EpiT) is used to detect T abuse in sport. Also, plasma or urinary concentrations of EpiT have been measured to assess testicular steroidogenesis during hormonal male contraception. Further investigations are required to evaluate the relative contributions of the testis and adrenal to EpiT production. To this purpose, we have compared basal urinary EpiT glucuronide and plasma EpiT and the response to synthetic adrenocorticotrophic hormone (ACTH) stimulation between eugonadal and hypogonadal men. DESIGN AND SUBJECTS: The basal urinary excretion rate of EpiT glucuronide was determined in 34 eugonadal men. Six men, clinically diagnosed as hypogonadal, and 6 out of the 34 eugonadal men previously described, received an intramuscular injection of synthetic ACTH depot (1 mg) at 0800 h on two consecutive days. Blood samples were collected prior to and then at 1.5, 8, 24, 25.5, 32 and 48 h with respect to the first administration (0 h). 24-h urine specimens were collected from 0800 h on days 1 and 2 (baseline) and 3 and 4 (stimulation). MEASUREMENTS: Plasma EpiT, T and cortisol were measured by RIA and urinary EpiT and T, following glucuronide hydrolysis, by gas chromatography-mass spectrometry (extract combines aglycones with a minor amount of urinary free steroids). RESULTS: Basal excretion rates of EpiT glucuronide in eugonadal men (range: 62-751 nmol/24 h) were considerably greater than in hypogonadal men (range: 3-34 nmol/24 h). Mean basal plasma EpiT in eugonadal men (1.32 +/- 0.08 nmol/l) were greater than in hypogonadal men (0.68 +/- 0.04 nmol/l). In each group, synthetic ACTH stimulation increased plasma cortisol 4-fold. In eugonadal men, plasma and urinary EpiT were unchanged whereas plasma and urinary T glucuronide decreased in response to ACTH. In hypogonadal patients, ACTH increased plasma and urinary EpiT while plasma T remained unchanged. CONCLUSION: The testes are the major source of epitestosterone, the adrenal contribution being relatively modest. Following adrenal stimulation, urinary epitestosterone glucuronide increases considerably in hypogonadal men but this increase is masked in eugonadal men because testicular production is probably suppressed by the ACTH induced rise in cortisol. Activation of the adrenal cortex results in no change or only a small decrease in the urinary T/EpiT ratio in eugonadal men. PMID- 10468934 TI - Amniotic fluid levels of dimeric inhibins, pro-alpha C inhibin, activin A and follistatin in Down's syndrome. AB - OBJECTIVE: In the second trimester of pregnancy, inhibin A is significantly increased in maternal serum and decreased in amniotic fluid in Down's syndrome pregnancies compared to normal. We wished to further evaluate the levels of inhibin A, inhibin B, pro-alpha C inhibin, activin A and the binding protein follistatin in amniotic fluid in Down's syndrome and control pregnancies. DESIGN: Case-matched control study. PATIENTS: 29 Down's syndrome and 290 chromosomally normal control pregnancies were identified from records and amniotic fluid, collected at second trimester amniocentesis, retrieved from routine storage for analysis. MEASUREMENTS: Inhibin A, inhibin B, pro-alpha C inhibin, total activin A and follistatin were measured using sensitive and specific enzyme linked immunosorbent assays. RESULTS: The median (10th-90th percentiles) amniotic fluid inhibin A level in the control pregnancies increased from 334 (122-553) ng/l at 14 weeks' to 695 (316-1475) ng/l at 19 weeks' gestation. The corresponding figures for inhibin B and the alpha-subunit precursor inhibin pro-alpha C were 632 (185-1354) and 2062 (1237-3381) ng/l, respectively at 14 weeks' and 2439 (748 5307) and 3115 (2021-6567) ng/l, respectively at 19 weeks' gestation. Total activin A increased from 3795 (1554-5296) at 14 weeks' to 5086 (3059-8224) at 18 weeks' gestation. Expressed as multiples of the median (MoM) the median (95% CI) amniotic fluid levels of inhibin A, inhibin B, pro-alpha C inhibin and acitivin A in the Down's syndrome samples were 0.77 (0.59-0.85), 0.94 (0.63-1.23), 0.77 (0.49-0.84) and 0.77 (0.53-0.87), respectively. Compared to controls the levels of inhibin A, pro-alpha C inhibin and activin A were significantly lower in Down's syndrome pregnancies (P < 0.01, Mann-Whitney U test). Follistatin levels in the controls declined slightly from 2106 (1421-3538) ng/l at 14 weeks' to 1600 (1281-2543) ng/l at 18 weeks' gestation. Levels in the Downs' syndrome pregnancies were similar to controls. CONCLUSIONS: The data suggest that the production, secretion or metabolism of the inhibin alpha- and beta A-subunits is altered in Down's syndrome pregnancies in the second trimester. PMID- 10468935 TI - Cushing's syndrome secondary to ectopic ACTH secretion from metastatic breast carcinoma. AB - Breast carcinoma is a rare cause of ectopic ACTH syndrome. There are only two previously reported cases in which ACTH secretion is documented. We describe the case of a 56-year-old woman who presented with clinical and biochemical features of ectopic ACTH syndrome in the setting of metastatic breast carcinoma. Despite aggressive management of her ectopic ACTH syndrome, her course was complicated by opportunistic infection, respiratory failure and death. Immunostaining of the breast metastases for ACTH was positive and in situ hybridization revealed proopiomelanocortin gene expression. This is the first reported case of ectopic ACTH syndrome associated with metastatic breast cancer in which the technique of in situ hybridization has been used to confirm the breast cancer metastases as the source of ectopic ACTH secretion. PMID- 10468936 TI - Prevalence and clinical usefulness of thyroid antibodies in different disease states of the thyroid. PMID- 10468937 TI - An unusual cause of hyponatraemia. PMID- 10468938 TI - Growth hormone and exercise. AB - This review summarizes the interactions between GH and exercise. Not only does exercise have profound effects upon the GH/IGF-I axis per se, but there is increasing evidence that such physiological perturbations might be influential in the performance responses to repeated training. However, the effects of systemic administration of rGH in restoring exercise capability and muscle strength in GH deficient adults and the ergogenic benefits of GH doping amongst athletes remain unproven. What is certain is that these issues will be of increasing relevance to clinical endocrinologists. PMID- 10468939 TI - Casting new light on the clinical spectrum of neonatal severe hyperparathyroidism. PMID- 10468940 TI - Inhibin B levels in azoospermic subjects with cytologically characterized testicular pathology. AB - BACKGROUND AND OBJECTIVE: Inhibin B, a heterodimeric glycoprotein of gonadal origin, is the most important circulating form of inhibin in human males and an inverse relationship between inhibin B and FSH plasma levels was been recently observed. Azoospermia represents the end-point of different kinds of testicular damage, ranging from a normal spermatogenic pattern (obstructive forms) to the complete absence of germ cells (Sertoli Cell Only Syndrome, SCOS). Furthermore, azoospermia may be related to maturational disturbances at different levels (spermatogonial, spermatocytic, spermatidic). To better define the relationship between testicular damage and inhibin levels and to evaluate the diagnostic value of this hormone in the management of subjects with azoospermia, we performed specific inhibin B assays in a group of azoospermic subjects affected by different kinds of testicular pathology. PATIENTS: Eighty-nine azoospermic men were studied by testicular ultrasound examination, fine needle aspiration of the testes and hormonal parameters (FSH, LH and testosterone, inhibin B). Thirty normozoospermic subjects were considered as controls for seminal and hormonal parameters. DESIGN AND RESULTS: On the basis of cytological analysis five different testicular appearences were identified in azoospermic patients: (i) Sertoli cell only syndrome (SCOS); (ii) Severe hypospermatogenesis; (iii) Spermatogonial and/or spermatocytic arrest; (iv) Spermatidic arrest; (v) Normal germ line (obstructive forms). No difference in LH and testosterone levels was found among the different groups. A significant negative correlation was present between inhibin B and FSH both in azoospermic men (r = - 0.503, P < 0.0001) and normozoospermic controls (r = -0.361, P < 0.05). Groups characterized by obstructive azoospermia and spermatidic arrest showed inhibin B concentrations similar to normozoospermic subjects (130.7 +/- 73.5, 160.3 +/- 35.1 and 148.5 +/- 46.8 ng/l, respectively), while groups characterized by SCOS, severe hypospermatogenesis and spermatogonial and/or spermatocytic arrest showed mean inhibin B concentrations significantly lower than controls (56.5 +/- 56.0, 57.9 +/- 31.2; 48.9 +/- 16.7 ng/l, respectively). In the group of SCOS, 8 out of 42 subjects (19.0%) showed inhibin B concentrations within the normal range despite high FSH levels. CONCLUSIONS: This study demonstrated that, in humans, spermatids play an important role in the mechanism regulating inhibin B secretion by Sertoli cells. The significance of this hormone as a diagnostic parameter in azoospermic patients does not seem to be specific because it does not permit discrimination between obstructive forms or spermatidic arrest. Furthermore, despite an evident clinical, cytological and hormonal pattern for SCOS, inhibin B levels are normal in some of these patients. The significance of this latter result remains to be elucidated. PMID- 10468941 TI - GH replacement in 1034 growth hormone deficient hypopituitary adults: demographic and clinical characteristics, dosing and safety. AB - OBJECTIVE: Long-term experience of growth hormone (GH) replacement therapy in a large population of hypopituitary adults with GH deficiency (GHD) is limited, and safety surveillance is clearly essential. KIMS, the Pharmacia & Upjohn International Metabolic Database, is a long-term, open, outcomes research programme of hypopituitary adult patients with GHD who are treated in a conventional clinical setting. PATIENTS: The present analysis encompasses data from 1034 hypopituitary adult GHD patients treated with GH for a total of 818 patient years. RESULTS: Prior to GH therapy, the KIMS patient population exhibited an increased prevalence of obesity, diabetes mellitus (in females) and hyperlipidaemia, compared with normal populations described in published studies. Quality of life, assessed using a disease-specific questionnaire (QoL-AGHDA), was also reduced in KIMS patients. The maintenance dose of GH was significantly higher in patients who were receiving GH prior to enrolment into KIMS (non-naive patients) compared with patients who commenced GH at the time of enrolment (naive patients). In addition, dose of GH correlated significantly with body weight in the former group of patients. Analysis of serum levels of IGF-I indicated that overtreatment with GH was markedly more common in non-naive than in naive patients. The frequency of adverse events in KIMS patients was no higher than that reported in patients receiving placebo in previous clinical trials. Recurrence of pituitary or CNS tumours was reported in six patients, a rate consistent with data from control series. Three deaths were reported, none of which was obviously associated with GH treatment. CONCLUSIONS: Our data, drawn from a large population of hypopituitary adults treated with GH for a total of more than 800 patient years, confirm previous reports that untreated GHD in hypopituitary adults is associated with a number of important clinical problems. In addition, the results suggest that there has been a shift in recent years from determination of GH dose on the basis of body weight to dose titration of individual patients, and indicate that the latter technique has important advantages. The data provide further evidence that GH replacement therapy is well tolerated in adults. However, it is possible that some adverse events may not become evident over the time scale covered by the present analysis, and continued surveillance therefore remains mandatory. PMID- 10468942 TI - Correlation between urinary cortisol and 6-sulphatoxymelatonin rhythms in field studies of blind subjects. AB - OBJECTIVE: Blind individuals have different types of circadian rhythm disorders. In order to study these in the field reliable markers of circadian phase are required. The aim of the present study was to determine the usefulness of urinary cortisol as a marker rhythm in field studies. This was assessed by investigating the relationship between the cortisol rhythms and the previously determined melatonin rhythms from a large sample of blind people with different types of circadian rhythm disorders. DESIGN: Field study SUBJECT: Registered blind subjects (n = 49) classified as having light perception or better (n = 19, 12 men, 7 women, aged 23-61 years) or having no conscious perception of light (n = 30, 24 men, 6 women, aged 19-72 years) were studied in their normal environment. MEASUREMENT: Sequential 4-hourly urine samples (plus an overnight sample) were collected for 48 h each week for 3-5 weeks. Urinary cortisol and 6 sulphatoxymelatonin were measured by radioimmunoassay. RESULT: Irrespective of the type of circadian rhythm (entrained or free-running), there was a significant correlation between the characteristics of the 6-sulphatoxymelatonin and cortisol rhythms in the blind subjects. CONCLUSION: Urinary cortisol is recommended as a useful marker of circadian phase in field studies in addition to urinary 6 sulphatoxymelatonin measurements. PMID- 10468943 TI - Constitutional delay in growth and puberty (CDGP) is associated with hypoleptinaemia. AB - OBJECTIVE: Serum leptin concentrations are higher in early adolescence compared with childhood and may play a facilitatory role in pubertal development. Constitutional delay in growth and puberty (CDGP) is a disorder of the tempo of physical maturation and may be associated with relative hypoleptinaemia. We have therefore compared serum leptin concentrations in normal boys with those in boys exhibiting constitutional delay of growth, controlling for pubertal status, age and body mass index (BMI). PATIENTS: 23 boys with constitutional delay in growth (n = 17, prepubertal) and puberty (n = 6, early pubertal) and 88 normal boys (n = 64 prepubertal, n = 24 Tanner stage 2). MEASUREMENTS: Serum leptin was measured in a single, non-fasted morning serum sample by radioimmunoassay. Using the data from normal boys, leptin standard deviation scores (sds) were calculated, to account for the independent influences of age and body mass index (BMI) sds. Both chronological age and bone age were used in the calculation of leptin sds in those with delay. RESULTS: BMIsds was significantly lower in prepubertal delays compared with controls but was not different in pubertal subjects. Leptin concentrations were higher in early puberty compared with prepuberty (P < 0.001) in normal boys but were not significantly elevated in pubertal boys with delay compared with prepubertal delays. Although leptin sds, for both chronological age and bone age, was similar in prepubertal controls and delays, it was significantly lower in the pubertal delay group (P < 0.05). In controls leptin sds did not correlate with age. However, in delays leptin sds was negatively correlated with both chronological age (r = - 0.43, P < 0.05) and bone age (r = - 0.68, P < 0.01), indicating that in the older delays leptin levels were lower than expected given their age and BMIsds. CONCLUSIONS: These data indicate that significantly higher leptin levels in Tanner stage 2 compared with prepuberty are not a prerequisite for progression into puberty. However, the absence of elevated leptin concentrations may be associated with delayed puberty in boys. PMID- 10468944 TI - Increased MEN1 mRNA expression in sporadic pituitary tumours. AB - BACKGROUND: The MEN1 gene on chromosome 11q13 encodes a tumour suppressor gene, mutations in which cause multiple endocrine neoplasia (MEN) type 1 syndrome. Loss of heterozygosity (LOH) at this locus is a common finding amongst sporadic pituitary tumours. We have therefore screened the MEN1 gene for mutations in sporadic pituitary tumours and, as the gene is a putative tumour suppressor, have quantified mRNA expression in tumorous and normal pituitaries to assess the role of MEN1 in pituitary tumorigenesis. SUBJECTS AND DESIGN: Thirty-one nonfunctioning pituitary tumours, 8 GH secreting, 2 TSH-secreting tumours and 1 corticotrophinoma have been assessed for the presence of MEN1 mutations, to examine the hypothesis that MEN1 mutations may contribute to the pathogenesis of sporadic pituitary neoplasms. In addition, quantitative changes in the pretranslational expression of the tumour suppressor gene MEN1 have been determined in 42 pituitary tumours and 6 normal pituitaries using semiquantitative reverse transcriptase PCR. RESULTS: No novel or previously published mutations were apparent in the MEN1 coding regions of any tumours studied, although several polymorphisms were identified. Transcriptional changes of the gene, assessed by semiquantitative RT-PCR, indicated that nonfunctioning and GH-secreting pituitary tumours are associated with significantly increased pretranslational expression of the MEN1 gene. In addition, the single corticotrophinoma showed increased expression compared to normal, as did one of the two TSH-omas. CONCLUSION: Coding mutations of the putative tumour suppressor gene MEN1 are unlikely to contribute to pituitary tumorigenesis in sporadic nonfunctioning, GH-secreting and TSH-secreting adenomas. Changes in pretranslational expression of MEN1 were observed in pituitary tumours, suggesting that changes in the level of MEN1 expression, rather than coding changes, may be of functional importance in influencing sporadic pituitary tumorigenesis. PMID- 10468945 TI - Reduced foetal growth and growth hormone secretion in adult life. AB - OBJECTIVES: Recent studies suggest that growth restriction or other adverse influences acting in utero or during early infancy lead to permanent alterations in growth hormone (GH) secretion. As GH secretion is known to predict cardiovascular risk, alterations in GH may contribute to the association between reduced foetal growth and cardiovascular disease. We have therefore assessed the relationship between birth size and GH secretion in a prospective study of young adults whose birth size was recorded and who have had their current blood pressure and glucose tolerance measured. DESIGN: Prospective cohort study PATIENTS: 153 healthy men and women, aged 20-21 years. MEASUREMENTS: Subjects carried out a timed overnight urinary collection for analysis of GH excretion. Insulin sensitivity and insulin secretion were measured using the intravenous glucose tolerance test with minimal model analysis. Blood pressure, height, weight, usual level of exercise, smoking habits, alcohol consumption, and socio economic status were also recorded. RESULTS: GH excretion ranged from 0.01 to 41.8 microU per subject. It did not differ according to gender but was markedly reduced in obese subjects (P < 0.0001) Low birthweight was strongly associated with low GH excretion at age 20 years (P = 0.002). Low placental weight and short body length also predicted low GH (P = 0.02 and P = 0.04, respectively). These relationships were independent of other confounding factors including obesity. GH excretion was not independently related to current levels of blood pressure, insulin sensitivity or insulin secretion. CONCLUSIONS: Body size at birth predicts GH excretion in adult life. Low GH excretion in people who were small at birth may be one mechanism explaining their increased risk of cardiovascular disease. PMID- 10468946 TI - Hypothalamic derangement in traumatized patients: growth hormone (GH) and prolactin response to thyrotrophin-releasing hormone and GH-releasing hormone. AB - OBJECTIVE: To study the impact of severe head injury on both basal pituitary hormone secretion and the response to exogenous synthetic hypothalamic releasing factors (TRH and GHRH) in order to evaluate sequential changes in the central control of hypophyseal secretion in the days following head injury. DESIGN: Prospective clinical study PATIENTS: 21 comatose male patients with head injuries, each intubated and ventilated, intensively monitored and having no previous endocrine problems. MEASUREMENTS: AND RESULTS The GH and PRL responses to TRH (200 microg iv), and the GH and PRL responses to GHRH (50 microg iv) were evaluated, respectively, on the days 1 and 16 and on days 2, 7and 15 after admission. Daily blood samples were also collected for GH, PRL, TSH, T3 and T4 evaluation. In the basal samples taken on days 2, 7 and 15, IGF-I and cortisol were also determined. Nitrogen balance was assessed daily. On the day 1, TRH increased GH levels from 9.8 +/- 2.2 to 22.4 +/- 6.5 mU/l but failed to induce GH release on day 16. The PRL response to TRH was normal. The GH peak response to GHRH was normal on the day 2 (35.7 +/- 13.9 mU/l), but was increased on days 7 and 15 (68.3 +/- 10.7 mU/l on day 7; 73.8 +/- 9.2 mU/l on day 15, P < 0.01 vs. day 2). We found a significant PRL response to GHRH during the whole period of observation. In the daily evaluation, nitrogen balance was negative in all patients from the day 1 to 5. On average, all patients reached a positive nitrogen balance on the day 8. Compared to the day 2, a statistical increase in IGF-I concentration was observed on days 7 and 15. CONCLUSIONS: The evaluation of pituitary dynamics in the acute phase of a severe injury demonstrates an alteration of GH and PRL secretion, which correlate with the aminergic and/or peptidergic derangements. Taken together, our data suggest augmented tone of both GHRH and somatostatin in the very acute phase, while an imbalance of releasing factors is hypothesized in the following days. The metabolic consequences of this neuroendocrine pattern could be advantageous in the rapid recovery from the cascade of events produced by the trauma, as documented by the increase in IGF-1 levels and the positive nitrogen balance. PMID- 10468947 TI - Dose titration and patient selection increases the efficacy of GH replacement in severely GH deficient adults. AB - OBJECTIVE: Previous studies of GH replacement in adults have used unselected cohorts of GH deficient (GHD) adults and weight-based dosing regimens resulting in supraphysiological serum IGF-I levels and a high frequency of side-effects and withdrawal from these studies. By choosing patients with a high level of morbidity at baseline and using a low dose GH titration regimen we aimed to avoid over-replacement and increase the efficacy of treatment. DESIGN: An open study of GH replacement, initiating treatment with a dose of 0. 8 U/day and titrating the dose by 0.4 U increments to normalize the IGF-I SDS between - 2.0 and + 2.0 SD of the age-related normal range. PATIENTS: 65 severely GHD patients (peak GH < 9 mU/l to provocative testing), 25 males, of mixed adult and childhood-onset and mean age 38.7 (range 17-72) years. Inclusion criterion was that of subjectively poor quality of life on clinical interview. MEASUREMENTS: Height, weight, waist and hip circumference were measured to allow calculation of body mass index (BMI) and waist-hip ratio (WHR). Bone mineral density (BMD) was measured by dual X-ray absorptiometry (DEXA). Serum haemoglobin A1C (HbA1C), lipid profile and insulin like growth factor 1 (IGF-I) were measured. The Psychological General Well-Being Schedule (PGWB) and Adult Growth Hormone Deficiency Assessment (AGHDA) self rating questionnaires (SRQ) were used to assess quality of life. RESULTS: Baseline characteristics were consistent with those previously described in severely GHD adults; mean IGF-I SDS - 2.4 (+/- 2.7), BMI 28.8 (+/- 5. 4) kg/m2, total cholesterol 6.17 (+/- 1.2) mmol/l, reduced BMD z-scores at the lumbar spine (- 0.8 +/- 1.2) and femoral neck (- 0. 44 +/- 1.4), and SRQ scores considerably lower than reported in previous studies of GH deficient adults and normal controls. Following initiation of GH serum IGF-I SDS was increased significantly from baseline to a mean level of 0.15 +/- 2.7 (P < 0. 001) and 0.31 +/- 2.0 (P < 0.001) at three and eight months, respectively. The mean PGWB score increased from 59.7 +/- 19.9 to 75. 8 +/- 15.0 (P < 0.001) and 73.7 +/- 19.5 (P = 0.001) at three and eight months, respectively. An increase of 14 points represents the largest improvement in quality of life, using this index, that has been reported in GHD adults. The mean AGHDA score also demonstrated considerable improvement, falling from 15.3 +/- 6.0 to 10.4 +/- 6.2 (P < 0.001) and 9.8 +/- 6.5 (P < 0.001) at three and eight months, respectively. The changes observed in both the PGWB and AGHDA scores between baseline and at both three and eight months were shown to correlate significantly with the respective baseline score. A significantly greater improvement was observed in the PGWB following GH replacement in those with a baseline PGWB score of < 60 than in those with a score > 60. This observation was significant at both three (27.1 vs 6.7, P = 0.0001) and eight (25.6 vs 3.3, P = 0.0003) months. All PGWB subscales showed significant improvement though that of vitality was of greatest magnitude. A strong correlation was observed between the generic and disease-specific SRQ (r = - 0.73, P < 0.001). CONCLUSIONS: The observed improvement in quality of life in GH deficient adults is proportional to the degree of impairment before commencing therapy. The use of low-dose titration and selection of a population with greater morbidity reduces the occurrence of over-replacement and increases the efficacy of treatment. This allows direction of resources to those in greatest need. PMID- 10468948 TI - The influence of hydrocortisone substitution on the quality of life and parameters of bone metabolism in patients with secondary hypocortisolism. AB - OBJECTIVE: Hydrocortisone replacement regimes remain rather empirical and produce serum cortisol profiles very different from normal physiology. We have analysed the effects of different dosages of hydrocortisone (HC) replacement therapy on the health perception and general well-being of patients with secondary hypocortisolism. We also evaluated the effects of these regimens on bone metabolism. DESIGN: In a prospective randomized double-blind study, 3 groups of 3 patients were treated with 3 different dosages of HC (15, 20 and 30 mg/day), in different sequences, each sequence for two weeks. PATIENTS: Nine adult patients with complete secondary hypocortisolism. MEASUREMENTS: Serum cortisol, ACTH, aldosterone, renin, alkaline phosphatase, bone specific alkaline phosphatase, osteocalcin, PTH, C-telopeptides of type-I collagen, sodium, potassium, phosphate; urinary free cortisol, pyridinium cross-links, urine sodium, potassium and phosphate were measured at the beginning and after each week of the study. For quality of life assessment the patients completed three different questionnaires, the Basler Befindlichkeits-Skala (BBS), the Befindlichkeits-Skala (Bf-S), the Beschwerde-Liste (BL) each week. RESULTS: With increasing doses of 15, 20 and 30 mg hydrocortisone a significant increase of free urinary cortisol was achieved (298 +/- 26 nmol/day, 454 +/- 43, 819 +/- 59, respectively; P < 0.01). The mean scores of the psychological questionnaires did not change significantly during the whole study (BBS 81.8 +/- 3.9; 82.8 +/- 3.9, 83.6 +/- 3.9; Bf-S 15.9 +/- 3.4, 11.3 +/- 2.6, 12.5 +/- 2.8; BL 15.7 +/- 2.3, 14.4 +/- 2.5, 14.8 +/- 2.6, respectively). Osteocalcin decreased significantly (2. 3 +/- 0.49, 2.1 +/- 0.42, 1.8 +/- 0.38, P < 0.01) with increasing HC doses but remained within the normal range. The other investigated parameters were within or nearly within the normal range in all patients at the beginning and did not change during the study. CONCLUSION: Dosages of 15, 20 or 30 mg hydrocortisone/day have equivalent effects on quality of life in patients with secondary hypocortisolism. With 15 or 20 mg hydrocortisone/day the patients feel nearly as well and content as normal healthy individuals. Since long-term treatment with a high replacement dose of glucocorticoids (hydrocortisone 30 mg/day) induces bone loss, this risk can be avoided with a substitution dosage of 20 mg or even 15 mg hydrocortisone/day, without influencing the well-being of the patient. PMID- 10468950 TI - The impact of gender, puberty and body mass on reference values for urinary growth hormone (GH) excretion in normally growing non-obese and obese children. AB - OBJECTIVE: There is a lack of normal reference data on urinary GH (u-GH) excretion in children. We have investigated the impact of age, gender, pubertal development and body mass on reference values for u-GH excretion in normally growing non-obese and obese children. STUDY DESIGN: u-GH levels were evaluated in 1153 healthy normal children (aged 5-14) and in 684 obese children (body mass index, BMI: > 75th). u-GH levels (ng/8 h) were determined by ELISA as the mean value of three consecutive first morning voidings. RESULTS: Reference values (5 95th centile) for u-GH excretion in obese and non-obese children of both sexes are reported. In normal prepubertal children median u-GH levels were relatively stable and superimposable in the two sexes; subsequently, u-GH levels increased, reaching a peak value at 13 years in both sexes. Significant increments (P < 0.0001) in u-GH levels were shown at B2 for females and at G3 for males. A slight decline was evident at 14 years. In obese children, median u-GH concentrations were significantly lower than those recorded in normal children of prepubertal age and at all stages of puberty (except in females at B2), in spite of their comparable normal height. u-GH levels significantly increased at puberty also in obese children, although the pubertal rise was significantly (P < 0.001) lower (1.7-fold in both sexes) than that observed in normal children (2.5-fold in boys and 2.3-fold in girls). A multiple regression analysis showed that both chronological age (beta: 0.20), BMI (beta: - 0.11), gender (beta: - 0.04) and pubertal stage (beta: 0.25) contributed significantly to the physiological variation in u-GH levels (multiple R: 0.44, P < 0.00001). CONCLUSIONS: This study provides reference values for u-GH in normally growing non-obese and obese children, analysing the impact of gender, puberty and body mass on this parameter. In agreement with previous studies, which demonstrate blunted GH responses to provocative stimuli and reduced nocturnal GH concentration, obese children have significantly lower u-GH levels than age-matched normal weight children, both before and during puberty. PMID- 10468949 TI - Correlation between cortisol and insulin-like growth factor-binding proteins (IGFBPs) under physiological conditions in children. AB - OBJECTIVE: A positive correlation between 24-h spontaneous growth hormone (GH) and cortisol secretion was previously reported in children. This observation prompted us to examine the relationship between physiological diurnal cortisol variation and the levels of insulin-like growth factors (IGFs) and IGF-binding proteins (IGFBPs) under physiological conditions. DESIGN AND PATIENTS: Starting at 0800 h, blood was sampled every 20 minutes over 24 h for measurement of GH and cortisol concentration in nine non-GH- deficient boys as part of a protocol for the investigation of short stature. MEASUREMENTS: IGFBP-1 and insulin were measured in samples drawn every 4 h over the 24-h period while IGF-I, IGF-II, IGFBP-2 and IGFBP-3 were determined in samples collected at the end of the study. RESULTS: No correlation was observed between IGF-I or IGF-II and mean cortisol levels. IGFBP-1 concentrations showed a marked circadian variation that was superimposed on the circadian rhythm for cortisol while a significant positive correlation was found for single point measurements between IGFBP-1 concentrations and cortisol levels measured in the same sample (r = 0.53) or at the preceding 20 minutes (r = 0.43), 40 minutes (r = 0.47) and 2 h (r = 0.38), suggesting an interplay between cortisol and IGFBP-1. A negative correlation (r = - 0.54) was found between IGFBP-1 and insulin levels determined in the same sample. A negative correlation (r = - 0.93) was also found between IGFBP-2 levels and mean cortisol concentrations during the preceding 12 h. No correlation was observed between plasma IGFBP-3 measured by IRMA and mean cortisol levels. CONCLUSION: Our data indicate a clear correlation between cortisol and IGFBP-1 and IGFBP-2 levels. Thus, the interplay of spontaneous GH and cortisol secretion in children may involve changes in IGFBP-1 and IGFBP-2 levels. PMID- 10468951 TI - Determinants of longterm outcome of radioiodine therapy of sporadic non-toxic goitre. AB - OBJECTIVE: Radioiodine treatment is effective in reducing the size of sporadic nontoxic goitre, albeit at the expense of a high incidence of postradiation hypothyroidism. The decrease in goitre size, however, is not observed in all subjects, and little is known about recurrent goitre growth after 131I therapy. The aim of the present study was to evaluate which factors determine the longterm outcome of 131I treatment in patients with sporadic nontoxic nodular goitre, in terms of changes in both thyroid size and thyroid function. STUDY DESIGN: Retrospective follow-up study. PATIENTS: Fifty patients with sporadic nontoxic nodular goitre were evaluated who had been treated in our institution with 131I (mean dose 4.4 MBq/g thyroid) in the period 1988-95. Nine patients received a second dose of 131I and one a third. Median follow-up time was 41 months (range 24-115). MEASUREMENTS: Thyroid function was assesed by TSH and FT4 index, and thyroid volume by ultrasound in 46 patients, by scintiscan using the Himanka formula in three and by CT-scan in one. The response to treatment was defined as a decrease in thyroid volume of greater than 13% (i.e. the mean + 2SD of the coefficient of variation of volume measurements), and recurrent goitre as an increase in thyroid volume greater than 13% after an initial response. RESULTS: Goitre size decreased from a median value of 82 ml (range 17-325) to 37 ml (range 6-204) two years after 131I treatment, a median reduction of 49%. The decrease in goitre size was directly related to the dose of 131I (r = 0.50, P = 0.0003) and indirectly to baseline goitre size (r = - 0.35, P = 0.006). Seven patients (14%) were nonresponders, and four (8%) experienced recurrent goitre growth after 3-5 years. These 11 patients (22%) when compared to the remaining 39 responders (78%) had larger goitres with more often a dominant nodule, and had received a lower 131I dose. The efficacy of a second dose of 131I (median reduction in goitre size 37%) was comparable to the first dose. Hypothyroidism occurred in 24 patients (48%), mostly in the first two years after treatment; 11 had overt and 13 subclinical hypothyroidism. Kaplan Meier statistics indicated a probability of 58% for developing hypothyroidism after 8 years. Hypothyroid patients had a smaller initial goitre size and a higher prevalence of TPO antibodies and a family history of thyroid disease than the patients who remained euthyroid; the 131I dose did not differ between the two groups. CONCLUSIONS: The size of sporadic nontoxic goitres is reduced on average by 50% after a single dose of 4.77 MBq 131I/g thyroid. Independent determinants of the relative decrease in thyroid volume are administered 131I dose and initial goitre size. Nonresponders (14%) and those with late recurrence of goitre growth (8%) have larger goitres and more often dominant nodules than responders. Determinants of postradioiodine hypothyroidism (cumulative risk 58% after 8 years) are the presence of TPO antibodies, a family history of thyroid disease and a relatively small goitre. The implications of these findings are that the efficacy of a given 131I dose can be enhanced when administered at an earlier stage when the goitre is still smaller, albeit at the expense of an increased risk for developing hypothyroidism. PMID- 10468953 TI - Different immunohistochemical patterns of TGF-beta1 expression in benign and malignant adrenocortical tumours. AB - OBJECTIVE: Transforming growth-factor beta1 (TGF-beta1) influences a number of specific functions of adrenocortical cells in several animal species. The aim of our study was to evaluate by immunohistochemical analysis the presence and distribution of TGF-beta1 in normal adrenal tissue and in different adrenal tumours. PATIENTS: We analysed 8 functioning (5 adenomas and 3 carcinomas) and 15 non functioning (6 adenomas and 9 carcinomas) adrenal tumours and 6 normal adrenal glands. RESULTS: In normal adrenal glands, the glomerulosa and the reticularis zones displayed diffuse cytoplasmic staining, while the fasciculata zone was almost completely negative. Functioning adenomas displayed cytoplasmic staining restricted to compact cells while in nonfunctioning adenomas, prevalently composed by clear cells, no staining was observed. Overall, adrenal carcinomas were characterized by the lack of cytoplasmic positivity and by sporadic positive cells around vessels both in functioning and in nonfunctioning tumours. CONCLUSIONS: TGF-beta1 expression is associated with active steroid secretion in normal adrenal tissue, as well as in benign cortical adenomas, while this relationship is lost in primary adrenal malignancies. These data provide indirect evidence for a regulatory role played by TGF-beta1 on steroid secretory pathways. PMID- 10468952 TI - Reduced androgen levels in adult turner syndrome: influence of female sex steroids and growth hormone status. AB - BACKGROUND AND OBJECTIVES: In girls with Turner syndrome androgen levels are reduced. In order to assess androgen status in women with Turner syndrome, we compared untreated adult women with Turner syndrome with a group of normal women. In addition, the effects of female sex hormone replacement therapy and GH status on the levels of circulating androgens in Turner syndrome was examined. DESIGN: All patients were receiving female hormone replacement therapy (HRT), which was discontinued four months prior to the initial examination. Patients were studied before and during HRT. Following the initial evaluation, patients were given cyclical HRT for six months consisting of either oral substitution (17beta oestradiol with norethisterone from day 13-22), or transdermal oestrogen substitution (17beta-oestradiol) with 1 mg norethisterone administered orally from day 13-22. Control subjects were studied once in the early follicular stage of the menstrual cycle. SUBJECTS: The study group consisted of 27 (33.2 +/- 7.9 years) patients with Turner syndrome and an age matched control group of 24 (32.7 +/- 7.6 years) normal women. MEASUREMENTS: Body composition measures, SHBG, testosterone (T), free testosterone (FT), dihydrotestosterone (DHT), alpha-4 androstendione (A), dehydroepiandrosterone sulphate (DHEAS), 17beta-oestradiol (E2), oestrone (E1), oestrone sulphate (ES), 24 h integrated GH concentration (ICGH), insulin-like growth factor I (IGF-I), insulin-like growth factor binding protein (IGFBP-3) were determined at baseline and after six months in women with Turner syndrome, and at baseline in control women. RESULTS: Circulating levels of A, T, FT, DHT, and SHBG were reduced by 25-40% in comparison with age matched normal women. The level of DHEAS was normal. The level of E2 was undetectable and levels of E1 and ES were very low in untreated Turner women. Treatment with 17beta-oestradiol and norethisterone increased oestrogen to levels comparable to those of normal women, while further decreasing FT (P = 0.02), DHT (P = 0.04), and T (P = 0.1). In untreated women with Turner syndrome IGF-I correlated significantly with DHEAS (R = 0.503, P < 0.01), while in normal women IGF-I correlated with A (R = 0.637, P < 0.01), T (R = 0.536, P < 0.01), and FT (R = 0.700, P < 0.01). During hormonal replacement in women with Turner syndrome IGF-I correlated significantly with DHEAS (R = 0.547, P < 0.01). Employing multiple regression analysis IGFBP-3, ICGH, DHEAS and fat free mass explained 85% (adjusted R = 0.92, P < 0.0005) of the variation in the level of IGF-I in untreated Turner syndrome. In treated Turners IGFBP-3, ICGH, SHBG, T, and FT explained 78% (adjusted R = 0.88, P < 0.0005). In controls IGFBP-3, SHBG, BMI and age explained 74% (adjusted R = 0.86, P < 0.0005) of the variation in IGF-I, while GH status did not contribute at all. CONCLUSION: The present study shows that many adults with Turner syndrome have reduced levels of circulating androgens, compared with an age-matched group of normal women. Conditions associated with Turner syndrome such as increased prevalence of sexual problems, reduced bone mineral content, osteoporosis, and an increased incidence of fractures and alterations in body composition could perhaps be alleviated or abolished by substitution with a low dose of androgens. Treatment with female hormonal replacement therapy is associated with a decrease in testosterone, free testosterone and dihydrotestosterone, possibly mediated by the androgenic effect of norethisterone. Furthermore significant differences in sex steroid levels, GH status and indices of body composition can be compatible with comparable levels of IGF-I in two very different groups of individuals. PMID- 10468954 TI - Fetal pituitary negative feedback at early gestational age. AB - We describe an early prenatal diagnosis and the successful treatment of fetal Graves' disease from transplacental transfer of maternal thyroid stimulating autoantibodies (TSAb). The diagnosis of fetal thyrotoxicosis was made by umbilical cord sampling (UBS) at 20 weeks gestation, based on suppressed TSH with elevated FT4 levels. Therapy with propylthiouracil (PTU) improved fetal thyroid function tests as well as the clinical signs of fetal Graves' disease. Three more UBS were conducted before delivery indicating persisting mild fetal hyperthyroidism. Undetectable concentrations of thyrotrophin in fetal serum in the presence of markedly elevated FT4, suggests pituitary negative feedback at as early as 20 weeks gestation. Amniotic fluid thyrotrophin levels were measured at 20,24 and 26 weeks and were shown to correlate better with (elevated) maternal rather than (suppressed) fetal TSH values; therefore, we believe that amniotic fluid thyrotrophin measurement is unreliable for prediction of fetal thyroid status. Our observation is the first documentation of an intact feedback mechanism so early in fetal development and it suggests that pituitary maturation occurs earlier than previously believed. PMID- 10468955 TI - Optimization of glucocorticoid replacement therapy: the long-term effect on bone mineral density. PMID- 10468956 TI - The entero-insular axis in glucagonoma. A report of 2 cases. PMID- 10468957 TI - How do levels of (endogenous) glucocorticoids, interleukin-10 and interleukin-12 relate to multiple sclerosis relapse before, during and after pregnancy? PMID- 10468958 TI - Apoptosis in neuroendocrine tumours. AB - Acquired resistance to apoptosis in neuroendocrine tumours (NETs) may promote clonal expansion and enhance the likelihood that subsequent mutations lead to growth or persistence of the neoplastic clone. Recent studies have demonstrated that deregulation of programmed cell death may be a critical component in multistep tumourigenesis of NETs and that the frequent expression of the Bcl-2 oncoprotein in these tumours may contribute to their pathogenesis. The genetic complementation of simultaneously deregulated Bcl-2 and c-Myc may be implicated in the multistep tumourigenesis of human NETs. Furthermore, because the efficacy of cytotoxic chemotherapy relies on its ability to induce programmed cell death, resistance to apoptosis typically correlates with chemoresistance, a phenomenon that is typical in NETs. Consideration of how oncogenes affect rates of cell death, in addition to augmenting growth, has already provided valuable insights into the biology of cancer. Understanding the molecular and cellular features of this process may enable the development and application of more effective and potentially curative treatment strategies in which the induction of programmed cell death is an integral component. PMID- 10468959 TI - Practice guidelines for the detection and management of thyroid dysfunction. A comparative review of the recommendations. PMID- 10468960 TI - Mapping of a novel tumour suppressor gene with a role in neuroendocrine tumours. PMID- 10468961 TI - Growth hormone replacement therapy in adults with growth hormone deficiency improves vascular reactivity. AB - OBJECTIVE: Patients with adult growth hormone (GH) deficiency are thought to be of increased risk of cardiovascular disease. Impaired vascular reactivity to endothelium derived nitric oxid (NO) is an early event in the development of atherosclerosis. In order to detect a possible effect of GH on vascular endothelium we examined forearm vasodilator responses in 8 patients with adult GH deficiency before and after 3 months GH replacement therapy. METHODS: Forearm blood flow studies were performed using venous occlusion plethysmography. Blood flow was measured at baseline and during intra-arterial infusions of 3 cumulative doses (7.5, 15 and 30 microg/minutes) of acetylcholine chloride and of sodium nitroprusside (1, 3 and 10 microg/minutes). Fasting blood samples were collected for measurement of lipid profile, Haemoglobin A1C (HbA1C), glucose, IGF-I and insulin. RESULTS: GH replacement therapy significantly increased IGF-I concentrations and tended to increase fasting insulin concentrations (IGF-I: 72.7 +/- 12.4 vs. 130.8 +/- 18.5 microg/l, P < 0.001; fasting insulin: 14.3 +/- 3.4 vs. 32.9 +/- 18.6, mU/l, P = 0.06). Fasting lipid profile, glucose and HbA1C did not significantly change. Blood flow responses to acetylcholine were significantly greater after GH replacement therapy (10.3 +/- 1.0 vs. 17.6 +/- 2.5 ml/minutes/100 ml for the highest dose, P < 0.03). There was a strong tendency to increased blood flow response to nitroprusside after GH therapy (10.7 +/- 1.2 vs. 17.5 +/- 1.7 ml/minutes/100 ml for the highest dose, P = 0.06). CONCLUSION: These findings suggest that GH replacement therapy may have a beneficial effect on endothelium function which is independent of quantitative changes in fasting lipid profile. PMID- 10468962 TI - Frequent loss of heterozygosity at chromosome 3p14.2-3p21 in human pancreatic islet cell tumours. AB - OBJECTIVE: Pancreatic islet betacell tumours occur either sporadically or as part of inherited neoplastic syndromes, most commonly multiple endocrine neoplasia (MEN) type 1. Recently, a transgenic mouse model has been established in which the expression of the SV40 large T antigen was targeted to betacells by the rat insulin promoter, leading to the development of multiple pancreatic betacell tumours. In the advanced stages of tumour evolution, these tumours exhibited a high prevalence of loss of heterozygosity (LOH) on mouse chromosomes 9 and 16, at regions syntenic with regions 3q, 3p21, 6q12, 15q24 and 22q of the human genome. DESIGN: Loss of heterozygosity in human islet cell tumours was analysed in a PCR based approach at regions of the human genome syntenic with the mouse loci linked to pancreatic betacell tumours as well as the MEN1 gene on chromosome 11q13. These included 35 microsatellite markers in the human chromosomal regions 3q, 3p21, 6q12, 11q13, 15q24 and 22q. PATIENTS: 21 patients diagnosed with insulinoma were analysed. Histologically, 16 tumours were benign, while 5 were malignant insulinomas. RESULTS: Thirteen of 21 (62%) tumours were found to have loss of genetic material on chromosome 3. The shortest region of overlap implicated a deletion at 3p14.2-3p21 region, corresponding to the marker D3S1295. We did not detect a substantial frequency of LOH in the other syntenic regions, except for the region of MEN 1 gene on 11q13 found to be deleted in 6 (29%) cases, including 3 of 4 tumours from MEN 1 families. Deletions of 3p14. 2-3p21 were observed in 8 of 15 (53%) benign tumours, and in 5 of 6 (83%) malignant neoplasms. CONCLUSIONS: These results indicate the high frequency of 3p14.2-3p21 deletions in human pancreatic betacell neoplasms. These finding suggest the presence of a tumour suppressor gene in this region, that may be important in the microevolution of these tumours towards malignancy. PMID- 10468963 TI - Evaluation of bone metabolism after the use of an inhaled glucocorticoid (flunisolide) in patients with moderate asthma. AB - OBJECTIVE: We have investigated the effects of the inhaled corticosteroid flunisolide on bone metabolism and adrenal function in patients with moderate asthma. SUBJECTS AND DESIGN: Twenty ambulatory patients (13 females, 7 males, mean age +/- SD of 36.4 +/- 12.4 years) with moderate asthma were recruited. None had taken corticosteroids for at least 1 month. Flunisolide 500 microg was given twice a day for 10 weeks, without any other medication. Blood and urine were collected before and at the end of treatment course. Cortisol (basal and 1 h after ACTH 250 microg i.v.) was measured to evaluate adrenal function. A peak cortisol response of 496 nmol/l was considered an adequate response. Serum ionized calcium, intact PTH, plasma osteocalcin (OC) and urinary pyridinoline (Pyr) and deoxy-pyridinoline (D-Pyr) were measured to evaluate bone metabolism. Wilcoxon paired test was performed for statistical analysis. Results are expressed as mean +/- SD. RESULTS: In most patients (85%), there was no difference after treatment with flunisolide on basal and stimulated cortisol levels. We found a significant decrease of OC (3.55 +/- 1.42 to 2.97 +/- 1.05 nmol/l) and Pyr (66.4 +/- 20.0 to 59.5 +/- 24.9 pmol/micromol creatinine) levels after treatment (P < 0.05). We also observed a positive correlation between the variations seen in pre and post treatment values of OC and Pyr/D-Pyr. CONCLUSIONS: The use of inhaled flunisolide 1000 microg/day for 10 weeks had no suppressive effect on adrenal function in the majority of asthmatic patients studied. However, the effects seen on bone and mineral metabolism, evidenced by the significant fall in osteocalcin and pyridinoline levels, may indicate a possible systemic effect of this drug. Clinical consequences of long-term treatment with flunisolide need to be further evaluated. PMID- 10468964 TI - Short-term leptin infusion does not affect circulating levels of LH, testosterone or cortisol in food-restricted pubertal male rhesus macaques. AB - OBJECTIVES: Although the adipocyte protein leptin has been implicated in the control of reproductive function in rodents, its role in primate reproductive physiology is poorly understood. Because primates in puberty show nighttime LH secretion and there is considerable evidence that the fertile state requires adequate nutrition, we reasoned that animals on the verge of reproductive competence would respond to leptin infusions by secreting LH. Food restriction reduces circulating leptin levels and slows or stops the GnRH pulse generator. Therefore, we examined the endocrine effects of leptin infusions in food restricted male pubertal primates during the night when they normally secrete LH. In addition, we investigated the effect of leptin on in vitro testosterone production by Leydig cells. SUBJECTS: Four pubertal male rhesus macaques (Macaca mulatta), 4-5.5 kg in weight (2.5-4-year-old) were examined in this study. Leydig cells from adult male rats were to investigate in vitro effects of leptin. DESIGN: To document that animals had entered puberty, blood samples were collected from each of the four animals at 15-minute intervals for 15 h both during the day and at night. Since at this age animals secrete LH mainly at night, blood samples were collected at 15-minute intervals from each of the four animals on two separate occasions for 15 h between 1500 and 0600h. During the experiment, animals were feeding from 0800 to 0830h, cages were completely cleaned of food at 0900h and the afternoon meal was not given to individual animals on the day they were studied. One of the studies served as the control (food restricted group) and during the other, 2 mg (n = 4) or 0.3 mg (n = 3) of recombinant human leptin was administered intravenously during 2000-0100h (food restricted plus leptin group). Blood samples (1 ml) were collected through the indwelling catheter and immediately transferred from the plastic syringe into chilled glass tubes containing 10 microl 14% EDTA. The samples were centrifuged at 5-h intervals and the plasma withdrawn and stored frozen at - 20 degrees C in polypropylene vials until assays were performed. MEASUREMENTS: Bioactive LH was determined and testosterone, cortisol and leptin were measured by radioimmunoassay. RESULTS: During daytime experiments in these animals, LH pulses were sometimes observed late in the day and generally continued for 12-15 h. Food restricted pubertal males showed delayed or absent LH pulses. Short-term leptin administration to food-restricted male rhesus macaques had no effect on LH, testosterone, or cortisol levels either during or after the infusion. Leptin also had no direct effect on basal or LH-stimulated testosterone production in Leydig cells. CONCLUSIONS: Our data support the notion that leptin is not the missing signal for the acute suppression of reproductive hormones secretion in food restricted primates. PMID- 10468965 TI - Effects of growth hormone replacement on physical performance and body composition in GH deficient adults. AB - OBJECTIVES: Adults with GH deficiency complain frequently of low energy levels resulting in a low perceived quality of life. Body composition is altered, with increased fat mass and decreased lean body mass, and muscle strength is reduced. The aims of this study were to determine the effects of GH replacement on physical performance and body composition in GH deficient (GHD) adults. STUDY DESIGN: The study consisted of a 6-month randomised, double-blind, placebo controlled study of the administration of GH (0.25 IU/Kg/week (0.125 IU/kg/week for the first four weeks)) followed by a 6-month open phase of GH therapy. PATIENTS: Thirty-five GHD adults (17F), mean age 39.8 years (range 21.1-59.9), on conventional replacement therapy as required. METHODS: Maximum aerobic capacity was measured using an incremental walking test to volitional exhaustion on a motorized treadmill. Quadriceps muscle strength was assessed by measuring maximum voluntary contractions and body composition by dual energy X-ray absorptiometry (DEXA). RESULTS: There were no statistically significant changes in quadriceps muscle strength between the GH and placebo groups. In both groups, there was a significant increase in quadriceps muscle strength compared to baseline during the double-blind period (GH group: P = 0.016; placebo group: P = 0.048). Compared to baseline, muscle strength was further improved in the GH treatment group after 12 months of treatment (P = 0.007). No further improvement was noted in the placebo group after 6 months on open GH treatment. In the placebo group, maximum aerobic capacity decreased during the placebo period (P = 0.017). No significant change was observed in the GH group. During open GH treatment the previously placebo treated group had a significant increase of maximum aerobic capacity (P < 0.049) whereas no significant improvement could be seen in the GH group. In the GH group there was a significant increase in lean body mass (P = 0.001) and a significant decrease in fat mass (P < 0.001). No statistically significant changes were noted in the placebo group: the differences in these changes between treatment groups were statistically significant (lean body mass: P = 0.009; fat mass: P < 0.001). The changes in body composition in the GH group during the 6 month placebo-controlled period were maintained during continued open treatment. Similar changes in body composition to those observed in the GH group during the 6 month placebo-controlled period were also seen in the placebo group once the patients received GH treatment. CONCLUSIONS: Our data show that GH replacement in GH deficient adults is associated with favourable changes in body composition, which could be important in the long term health outcome and physical activity of GH deficient patients. Our data support the concept that GH therapy alone, in the absence of some form of exercise programme, may increase the amount of lean tissue but not the quality or functional capacity of this tissue and it may be that training, in addition to GH therapy, may be necessary to significantly increase physical performance in these patients. We suggest that future trials with GH therapy and general approaches to the treatment of GH deficiency should include a planned activity programme as an approach to health improvement in these patients. PMID- 10468966 TI - Growth hormone status following treatment for Cushing's syndrome. AB - OBJECTIVE: Both pituitary surgery and radiotherapy for Cushing's disease can lead to growth hormone (GH) deficiency. Studies to date have, however, described the incidence of impaired GH secretion and not the incidence of severe GH deficiency following treatment of Cushing's disease. Furthermore, following cure of Cushing's disease and resolution of hypercortisolaemia, recovery of GH secretory status is seen, thus creating uncertainty as to the persistence of any documented GH deficiency. This study has two aims; to determine the incidence of severe persistent GH deficiency following treatment of Cushing's disease and to assess the time scale of any recovery of GH secretory status following surgical cure of Cushing's disease. DESIGN AND PATIENTS: The case notes of 37 patients either cured or in clinical and biochemical remission following treatment for Cushing's syndrome were reviewed to determine the incidence of severe GH deficiency. Of 34 patients with Cushing's disease, 20 were treated by pituitary surgery, and 14 with radiotherapy. Three patients with adrenal adenomas underwent unilateral adrenalectomy. MEASUREMENTS: GH secretory status was assessed by provocative testing using an insulin tolerance test (ITT, 85% of all tests), glucagon stimulation test (GST) or arginine stimulation test (AST). RESULTS: Thirty-six percent (5/14) of radiotherapy treated patients demonstrated severe GH deficiency at a mean time of 99 months following remission. Fifty-nine percent (10/17) of surgically treated patients assessed in the two years following remission demonstrated severe GH deficiency, whilst only 22% (2/9) of patients assessed beyond two years following remission demonstrated severe GH deficiency. This latter cohort is biased, with patients in whom severe GH deficiency had been demonstrated on earlier tests being over-represented. It is more accurate to estimate the incidence of persistent severe GH deficiency following surgically induced remission of Cushing's disease by incorporating data from patients in whom original testing demonstrated adequate GH reserve. Collating such data, 13% (2/15) of patients had persistent severe GH deficiency. Across all time periods five surgically treated patients demonstrated recovery of GH secretory status over a median time course of 19 months. In the surgically treated cohort, seven (35%) patients had anterior pituitary hormone deficits other than GH deficiency: 14% (2/14) of patients with normal GH secretory status at the last assessment, 83% (5/6) of patients with severe GHD at the last assessment. Of the 5 patients who demonstrated recovery of GH secretory status 40% (2) had additional anterior pituitary hormone deficits. Within the radiotherapy treated cohort 14% (2/14) of patients demonstrated additional anterior pituitary hormone deficits: 11% (1/9) of patients with normal GH secretory status and 20% (1/5) of patients with severe GH deficiency. None of the patients with adrenal adenomas treated by unilateral adrenalectomy demonstrated any abnormality of GH secretory status CONCLUSIONS: The incidence of severe persistent GH deficiency following surgically induced or radiotherapy induced remission of Cushing's disease is lower than has been suggested by previous studies, although these latter studies have assessed GH insufficiency and not severe GH deficiency. In the presence of additional pituitary hormone deficits severe GHD is common and is likely to be persistent. Recovery of GH secretory status is seen in a high proportion of patients reassessed, at a median time of 19 months following surgically induced remission of Cushing's disease. Thus, we recommend that definitive assessment of GH secretory status is delayed for at least two years following surgical cure of Cushing's disease. This has important implications for patients in whom GH replacement therapy is being considered. PMID- 10468967 TI - Salivary cortisol for screening of Cushing's syndrome in children. AB - OBJECTIVE: Cushing's syndrome (CS) is characterized by changes in diurnal cortisol variation and total or partial resistance to cortisol suppression by dexamethasone (DEX). Diagnosing CS is a challenge especially in childhood and requires differentiation from primary obesity. The aim was verify the efficacy of salivary cortisol in differentiating primary obesity from CS in children. SUBJECTS AND METHODS: We studied 11 patients with CS confirmed by standard laboratory investigation and surgical findings aged 1-16 years, and 21 age matched primary obese controls. Salivary samples were collected at 0900 h, 2300 h, and after an overnight DEX suppression test (20 microg/kg up to 1 mg). Salivary cortisol was measured by RIA. RESULTS: Diurnal variation of salivary cortisol levels was observed in all controls, as opposed to only 5 of 11 patients with CS. Suppression of salivary cortisol was detected in all controls but in no CS patient after the overnight DEX test. Mean salivary cortisol levels were higher in the CS than in the control group both at 2300 hours and at 0900 hours after DEX but no difference was observed at 0900 hours of the first day. The sensitivity and specificity of salivary cortisol at 0900 h, 2300 hours and after DEX in diagnosing CS were 27% and 95.2%, 100% and 95. 2%, and 100% and 95.2%, respectively. Sensitivity and specificity of 100% were obtained by combining the data for 2300 hours and after-DEX. CONCLUSION: The combination of salivary cortisol determination at 2300 hours and after-Dexamethasone is an easily performed and noninvasive method with high specificity and sensitivity for diagnosing Cushing's syndrome in children. PMID- 10468968 TI - Differences in the levels of TSH-binding inhibitor immunoglobulins in goitrous and agoitrous autoimmune thyroiditis after twelve months of L-thyroxine therapy. AB - OBJECTIVE: The aims were to study the prevalence of TSH binding inhibitor immunoglobulins (TBII) in newly diagnosed patients with autoimmune hypo thyroidism, to determine if clinical and biochemical parameters in these patients differed, and to study the course of these antibodies after 12 months of L thyroxine (LT4) therapy. SUBJECTS AND DESIGN: In a prospective study, 111 consecutive patients with newly diagnosed auto-immune hypothyroidism were enrolled. Patients were divided into groups according to the presence or absence of a goitre and TBII levels. Clinical and biochemical differences in these patients were analysed. Patients were then treated with L-thyroxine (LT4) for 12 months and changes in their TBII monitored. MEASUREMENTS: Free T4 and TSH levels were measured at baseline and then 6-weekly during the titration of LT4 doses. Once TSH levels had normalized, these measurements were performed 3 monthly. TBII levels were measured in patients at baseline and 12 months. Thyroid stimulating blocking antibody (TSBAb) levels were measured 12-18 months after initiation of LT4 therapy. RESULTS: Twenty patients were TBII-positive, 10 goitrous and 10 agoitrous. Agoitrous TBII-positive patients were found to have similar characteristics: these included TBII levels > 100 U/l, potent thyroid stimulating blocking antibody (TSBAb) activity and a tendency for TBII levels to remain unchanged or to rise after LT4 therapy. The degree of hypothyroidism seen in these cases was significantly greater than in all other groups. In contrast, goitrous TBII-positive patients were heterogenous, most did not have significant TSBAb activity and TBII disappeared in 5 out of 10 cases after a year of LT4 treatment. While TBII disappeared in 6 of the 20 patients after a year, remission of hypothyroidism occurred in only 3 cases. CONCLUSIONS: The degree of hypothyroidism in agoitrous TSH-binding inhibitor immunoglobulins-positive patients appears to be more severe than that seen in other forms of AIT. The differences in thyroid stimulating blocking antiboby activity and response to LT4 therapy in agoitrous and goitrous autoimmune thyroiditis suggest that the TSH binding inhibitor immunoglobulins in these patients are fundamentally different. The disappearance of TSH-binding inhibitor immunoglobulins in hypothyroid patients was not accompanied by the reversal of hypothyroidism in 50% of patients. PMID- 10468969 TI - Serum leptin levels in healthy ageing men: are decreased serum testosterone and increased adiposity in elderly men the consequence of leptin deficiency? AB - OBJECTIVE: The limited information on serum leptin levels in elderly men suggests the occurrence of an age-related decrease, with disruption of the relationship between fat mass and leptin levels. A relative leptin deficiency might thus be implicated in the increase of fat mass and decrease of serum testosterone levels in elderly men. Therefore, we have reevaluated the age-related changes in serum leptin levels and their relationship with adiposity and androgen levels in a large group of community dwelling men. SUBJECTS AND MEASUREMENTS: Serum leptin and androgen levels were measured in 271 healthy, ambulatory elderly men (median age 74 years), as well as in 61 middle-aged (median 43 years) and 40 young (median 25.5 years) controls. Adiposity was assessed by anthropometrical measurements (body mass index; BMI) and by estimation of fat mass by the bio impedance method. RESULTS: Serum leptin levels, whether or not adjusted for BMI, were found to increase with age, the values tending to level off after the age of 45 years, and were strongly correlated to BMI (r = 0.77) and fat mass assessed by the bio-impedance method (r = 0.81). Linear regression analysis showed a similar slope for the relationship between BMI and serum leptin in the three age groups. Multiple linear regression analysis indicated BMI, age and serum insulin, but not serum testosterone, as significant independent correlates of serum leptin. Serum (free) testosterone levels were negatively correlated with age and serum leptin, also after partialization for BMI: rank correlation coefficients vs. age and serum leptin, respectively, were - 0.20 (P < 0.001) and - 0.16 (P < 0.01) for total testosterone and - 0.60 (P < 0.001) and - 0.23 (P < 0.001) for free testosterone. Dehydroepiandrosterone sulphate (DHEAS) and leptin levels emerged as significant independent correlates in a multiple linear regression model for total serum testosterone; BMI and serum insulin became highly significant correlates in the same model when leptin was omitted from the independent variables. CONCLUSION: Ageing in men is accompanied by a rise of serum leptin levels with a maintained strong association between serum leptin and adiposity in elderly men. Testosterone does not appear to be a major determinant of serum leptin in healthy men, while leptin does emerge as a negative correlate of serum testosterone. Increased fat mass and decreased testosterone production in elderly men cannot be attributed to a relative leptin deficiency. PMID- 10468970 TI - Reduced human growth hormone (hGH) bioactivity without a defect of the GH-1 gene in three patients with rhGH responsive growth failure. AB - BACKGROUND: A GH deficiency-like phenotype with normal or high hGH secretion, pathologically low IGF-I serum levels, and catch-up growth under treatment with recombinant hGH is suggestive of the presence of biologically inactive hGH syndrome, whose presumably heterogenous molecular basis is substantially unknown. DESIGN: Serum samples from patients who fulfilled the above criteria and from controls with idiopathic short stature were measured by polyclonal hGH-RIA, Nb2 rat lymphoma proliferation assay, and hGH immunofunctional assay (IFA). If assays were suggestive of the presence of bioinactive GH, mutational analysis of the hGH 1 gene was performed. PATIENTS: Three patients were selected because of clinical and biochemical evidence. At the time of diagnosis mean age was 3.4 (2.2, 3.5, 4.5) years, mean height -3.5 (-2.8, -3.6, -4.2) SD score (SDS) and mean growth rate -1.5 (-1.4, -1.5, -1.6) SDS. Mean IGF-I serum levels were -1.9 (-0.7, -2.4, 2.5) SDS and mean IGFBP-3 serum levels -1.2 (-1.1, -1.2, -1.2) SDS. Stimulated and spontaneous GH peaks measured by a polyclonal radioimmunoassay were all above 14 microg/l. GHBP serum levels were normal, and antihGH antibodies were not detected. Therapy with rhGH was effective in causing catch-up growth of the three children with an initial mean growth rate of + 2.9 (+ 1.7, + 2.1, + 5.0) SDS, and normalization of IGF-I (mean: -0.66 SDS: -1.8, - 1.2, + 1.1 SDS) and IGFBP-3 serum levels (mean: + 0.81 SDS: -0.2, + 0.8, + 1.8 SDS). RESULTS: In comparison to controls, the patients' serum hGH levels were much lower when measured by Nb2 rat lymphoma cell proliferation bioassay (mean: -2.3 SDS, range: -1.7- -4.1) and by the immunofunctional assay (IFA) (-1.5 SDS, range: -0.2- -3.1) than by RIA. Retesting of two of the three patients including an one year break of therapy confirmed the rhGH dependence of growth in spite of normal endogenous GH secretion. Radioactive direct sequencing of both strands of PCR-amplified genomic DNA and cDNA excluded a GH-1 gene mutation in all three children. CONCLUSION: Mutations of the GH-1 gene are probably not the main genetic defect in children with biologically inactive hGH syndrome. Posttranslational processing of hGH might reduce the biological activity of the normal translation product. PMID- 10468971 TI - Mutation screening and isoform prevalence of the follicle stimulating hormone receptor gene in women with premature ovarian failure, resistant ovary syndrome and polycystic ovary syndrome. AB - OBJECTIVE: To determine whether mutations in the FSH receptor gene are associated with premature ovarian failure (POF) or resistant ovary syndrome (ROS) in women in the UK. To determine whether an allelic variant of the FSH receptor gene affects fertility parameters in women with polycystic ovary syndrome (PCOS). DESIGN: A mutation screen using DNA from women with POF and ROS. Restriction digest of amplified DNA from women with POF, ROS, PCOS and controls to determine allelic variant status. Fertility parameters were compared between allelic variant subgroups of women with PCOS. PATIENTS: The study population comprised 49 women with POF, 5 with ROS, 93 with PCOS and 51 controls. MEASUREMENTS: In women with PCOS, fertility and menstrual status was recorded and serum FSH and ovarian volume were measured. RESULTS: No mutation of the FSH receptor gene was found in women with POF or ROS. The allelic variant Thr307/Ser680 was found to be similarly prevalent in all study groups. The Thr307/Ser680 variant was found to have no phenotype in terms of fertility parameters in women with PCOS. CONCLUSIONS: Mutations of the FSH receptor gene are rare in women with premature ovarian failure or resistant ovary syndrome in the UK. Polymorphisms of the FSH receptor gene do not appear to have pathophysiological significance with regard to ovarian function. PMID- 10468972 TI - Type 1 multiple endocrine neoplasia (MEN1): contribution of genetic analysis to the screening and follow-up of a large French kindred. AB - OBJECTIVE: Multiple endocrine neoplasia type 1 (MEN1) is an autosomal genetic disorder, the clinical phenotype of which includes tumours of the parathyroids and/or anterior pituitary and/or endocrine pancreas. The genetic defect has been mapped to the chromosome 11q13 and the MEN1 gene has been recently identified, thus allowing genetic screening in affected kindreds. The aim of this study was to establish the usefulness of genetic screening in the follow-up of a large MEN1 kindred. PATIENTS: We describe a large kindred of 91 members, of whom 56 had clinical, biochemical and genetic screening. Twenty eight of them have been tested annually for the last 5 years. RESULTS: Although the precise mutation is still undetermined in this kindred, genotypic determination confirmed linkage with the MEN1 gene in affected members and excluded 28 members from annual testing. By drawing our attention to susceptible subjects, genetic screening improved the evaluation of age-related penetrance of the disease in the 3 generations from this kindred. For instance, annual screening showed conversion from unaffected to affected phenotype in 4 subjects aged 14, 14, 15, and 17 years. Moreover, genetics helped us to evaluate the specificity of clinical or biochemical markers, and thus to discard useless investigations. So far however, the genetics have not helped to explain the phenotypic heterogeneity and particularly low incidence of pancreatic tumours in this kindred. CONCLUSION: Genetic screening is very useful in detecting high risk individuals for MEN 1, since it avoids time-consuming and expensive investigations in non-affected subjects. By providing better understanding of the age-related penetrance of this syndrome, it improves the efficiency of screening. Genetic studies allow differentiation between clinical and biochemical features that are useful in follow-up and other confusing or unhelpful parameters. PMID- 10468973 TI - Association of the large multifunctional proteasome (LMP2) gene with Graves' disease is a result of linkage disequilibrium with the HLA haplotype DRB1*0304 DQB1*02-DQA1*0501. AB - OBJECTIVE: The large multifunctional proteasome (LMP) molecules are over expressed in thyrocytes, the target cells of Graves' disease, and the LMP genes are found within the MHC class II region. The LMP genes may therefore play a role in susceptibility to Graves' disease. The aim of this this study was to determine whether polymorphisms of the LMP genes, LMP 2 and LMP 7 are in linkage disequilibrium with Graves' disease. DESIGN: Target DNA was amplified using the polymerase chain reaction. The distribution of an Arg-His polymorphism in the LMP 2 gene and a G/T polymorphism in the LMP 7 gene, both of which lead to the presence of an HhaI restriction site, were investigated in a population based case control and family based study in patients with Graves' disease. PATIENTS: We obtained DNA from 306 patients with Graves' disease and 364 control subjects for the population based case-control study. In an independent family based study, DNA was obtained from 129 families including both parents, an affected sibling with Graves' disease and an unaffected sibling. All families, patients and control subjects were white caucasians. MEASUREMENTS: Frequencies of the alleles and genotypes of the LMP 2 and LMP 7 genes were compared between patients and control subjects using the chi2 test. Transmission of alleles from heterozygous parents to affected and unaffected offspring was assessed using the transmission disequilibrium test (TDT). RESULTS: In the case control study, no difference in allele or genotype frequency was seen between patients and control subjects at the LMP7 locus. At the LMP2 locus the R allele and the RH genotype were increased in subjects with Graves' disease when compared with control subjects (R allele: 36.3% vs. 29.5%, pc = 0. 0164; RH genotype: 56.5% vs. 45%, pc = 0.0102). However, the R allele was in linkage disequilibrium with the associated HLA DRB1*0304-DQB1*02-DQA1*0501 haplotype, delta = 0.102. Within the family based study, no preferential allelic transmission was seen from heterozygote parents to offspring at either loci. CONCLUSION: These results show that association of the LMP 2 locus with Graves' disease is due to linkage disequilibrium with the associated HLA haplotype DRB1*0304-DQB1*02-DQA1*0501. PMID- 10468974 TI - Macroprolactinaemia: validation and application of the polyethylene glycol precipitation test and clinical characterization of the condition. AB - OBJECTIVE: The clinical characteristics and natural history of macroprolactinaemia are yet to be fully established and there is currently no widely accepted simple in vitro test for routine identification of the condition. This study was designed to further validate the polyethylene glycol (PEG) precipitation test with assessment of percentage prolactin recovery (%R), for the detection of macroprolactinaemia and clinically characterize cases identified using the test. DESIGN: Analytical validation and application of the PEG precipitation test for diagnosis of macroprolactinaemia and a retrospective case notes review of some of the patients identified with the condition. PATIENTS: Sera from 188 patients with elevated serum prolactin (PRL) concentrations were screened for macroprolactinaemia. Seventeen of the patients who were under follow up at the local teaching hospital and were identified to have macroprolactinaemia were systematically characterized clinically. MEASUREMENTS: Prolactin was measured in sera, fractions from gel filtration chromatography and supernatant obtained after PEG precipitation, by the DELFIA fluoroimmunoassay. CLINICAL ENDPOINTS: Initial serum PRL and symptoms and their response to dopamine agonist therapy; imaging of the pituitary gland; serum PRL and symptoms after cessation of dopamine agonist therapy; course of serum PRL and symptoms without dopamine agonist therapy. RESULTS: The within-batch coefficient of variation (CV) of the PEG precipitation test ranged between 2.8 and 7.3% while the between-batch CV ranged between 3.4 and 9.7%. Intra-individual variability up to five months was 9.9%. A prolactin recovery 50%. The prevalence of macroprolactinaemia was 15% without sex difference. Clinical characterization of 17 patients showed modest initial serum PRL concentrations and there was imaging evidence of a coexisting pituitary adenoma in three of them. Dopamine agonist therapy resulted in substantial falls in serum PRL and this was associated with improvement or resolution of symptoms in some patients. Stoppage of dopamine agonist resulted in rebound hyperprolactinaemia in all cases. Spontaneous improvement in symptoms occurred in three patients but hyperprolactinaemia persisted in all those who were not on treatment. CONCLUSIONS: The PEG precipitation test with assessment of 'free' prolactin has been shown to be reproducible and sensitive for the detection of macroprolactinaemia. Many of the patients who had macroprolactinaemia presented with at least one symptom of the hyperprolactinaemic syndrome. Dopaminergic therapy may influence the course of both serum PRL and symptoms in some of these patients. PMID- 10468976 TI - Bone, chronic renal failure and GH treatment. PMID- 10468975 TI - Ovarian responses to hCG stimulation: insulin resistance/hyperinsulinaemia vs. insulin deficiency. AB - Polycystic ovary syndrome is a heterogeneous disorder characterized by signs and symptoms of hyperandrogenism and insulin resistance. We present the clinical and hormonal features in an adolescent girl who had distinct intervals of insulin deficiency and insulin resistance/hyperinsulinaemia. This case report confirms that insulin resistance/hyperinsulinaemia exacerbates ovarian hyperandrogenism. PMID- 10468977 TI - Are myocardial 'pseudovascular muscle tubes' present in acromegalic hearts? PMID- 10468978 TI - Thyroid function and autoimmunity during treatment with G-CSF. PMID- 10468979 TI - Neoplasia in patients with pituitary adenomas. PMID- 10468980 TI - Role of FSH in the regulation of spermatogenesis: clinical aspects. PMID- 10468981 TI - Non-islet cell tumour hypoglycaemia. PMID- 10468982 TI - The role of bilateral inferior petrosal sinus sampling in the investigation of ACTH-dependent hypercortisolism. PMID- 10468983 TI - The short Synacthen test: is less best? PMID- 10468984 TI - Reproducibility of the cortisol response to stimulation with the low dose (1 microg) of ACTH. AB - OBJECTIVE: Previous studies have shown that the rapid ACTH stimulation test using a low dose of 1 microg is more sensitive than that using 250 microg ACTH for detecting subtle cases of adrenal insufficiency. However, there are controversies for the reproducibility of the 1 microg-test. To evaluate the reproducibility of the 1 microg-test, we assessed both day-to-day and diurnal variations of cortisol responses to 1 microg ACTH injection. In addition, optimum sampling time for the 1 microg-test was also determined. SUBJECTS: AND DESIGN Eight healthy volunteers and five patients with secondary adrenal insufficiency were recruited. Healthy subjects were given 1 microg ACTH 3 times in the morning (0800 h) and 2 times in the afternoon (1600 h). Patients with adrenal insufficiency had 2-tests in the morning and 2 in the afternoon. Serum cortisol levels were measured every 10 minutes for 1 h after the injection. RESULTS: In healthy subjects, basal and peak serum cortisol levels were significantly higher in the morning (P < 0.05), whereas maximum cortisol increments were higher in the afternoon (P < 0.001). In patients with adrenal insufficiency, basal and peak serum cortisol levels in the morning were not different from corresponding values in the afternoon. Intra individual coefficient of variation (CV) of peak serum cortisol response to 1 microg ACTH ranged from 3.0 to 16.4% in healthy subjects and 10.0-34. 4% in patients. Also, there was a significant correlation between peak morning or afternoon cortisol levels after 1 microg ACTH injection given in different days in both healthy subjects and patients. Twenty-six of the 40 studies in healthy subjects showed peak response at 20 minutes, while nine showed it at 30 minutes Using the data acquired at 20 and 30 minutes, all 40 studies in healthy subjects showed normal results while none of 20 studies in patients was normal. CONCLUSIONS: We conclude that the cortisol response to 1 microg ACTH stimulation was reproducible in both healthy subjects and patients with secondary adrenal insufficiency. In order to assess adrenal function more accurately with the 1 microg ACTH stimulation test, serum cortisol should be measured before and 20 and 30 minutes after ACTH injection. PMID- 10468985 TI - Comparison of six months therapy with octreotide versus lanreotide in acromegalic patients: a retrospective study. AB - OBJECTIVE: We analysed the effects of 6-months' treatment with octreotide s.c. and lanreotide-SR on circulating GH and IGF-I levels in acromegaly. DESIGN: Open retrospective study. PATIENTS: Thirty-eight patients with active acromegaly (plasma IGF-I levels greater than 2 standard deviations for age-matched controls and increased serum GH levels not suppressible by oral glucose load) were studied. All patients received s.c. octreotide at a dose of 150-600 microg/day for six months as first therapy and subsequently, lanreotide i.m., 30-60 mg either at 14 or 10 day intervals, for 6 months. A 3 months' washout was applied before starting lanreotide treatment. MEASUREMENTS: Mean serum GH levels (from three samples), IGF-I, and clinical examination were performed before and 30, 60, 90 and 180 days after octreotide and lanreotide treatments. Safety tests, HbA1c and, thyroid function were evaluated every three months. RESULTS: Circulating GH and IGF-I levels were significantly reduced (P < 0.001) after one, three and six months of both octreotide and lanreotide treatment. The absolute values were lower and the percent decrease in serum GH levels obtained after octreotide treatment was significantly greater, at all scheduled assessments, than after lanreotide (P < 0.01). Serum IGF-I levels during octreotide were significantly lower only after the first month of therapy (P < 0.01). CONCLUSIONS: Our study shows that octreotide s.c. is able to induce an earlier reduction in IGF-I levels and a more marked reduction in GH levels than lanreotide. However, after six months of therapy the number of patients with safe GH levels and normal IGF-I age matched levels, was similar with both drugs. Therefore we suggest that octreotide treatment be preferentially used in the short-term presurgical treatment, while lanreotide can be used in chronic therapy when better compliance is necessary. PMID- 10468986 TI - A novel mutation in the TPO gene in goitrous hypothyroid patients with iodide organification defect. AB - OBJECTIVE: To screen and subsequently sequence the TPO gene for mutations in patients with congenital goitre, hypothyroidism and evidence for an organification defect (positive perchlorate discharge test). PATIENTS: We have studied seven hypothyroid and congenitally goitrous patients from three unrelated families. DESIGN AND MEASUREMENTS: We have measured serum thyroid hormone levels, 131I uptake, serum TSH and serum Tg concentrations. Denaturing gradient gel electrophoresis (DGGE) of PCR amplified genomic DNA was used to screen for mutations in the TPO gene. RESULTS: DGGE identified the presence of two frameshift mutations: a GGCC duplication in exon 8 (homozygous in one family and heterozygous in the other family) and a heterozygous insertion of a single nucleotide (C) at position 2505-2511 in exon 14. In addition, we have detected an alteration in exon 11, not yet described in the literature, derived from a single nucleotide substitution of a C to G at position 2008, altering the well-conserved amino acid domain among the peroxidases superfamily. This mutation in exon 11 was present in two families that showed heterozygous mutation for exon 8 or for exon 14. CONCLUSIONS: These results could support the hypothesis for a putative compound heterozygosity pattern in the affected patients. The altered phenotype (goitre and hypothyroidism since birth) seems justifiable in view of the possible inactivating character of this novel mutation in exon 11. PMID- 10468987 TI - Growth hormone (GH) replacement therapy reduces serum sialic acid concentrations in adults with GH-deficiency: a double-blind placebo-controlled study. AB - OBJECTIVES: Patients with adult GH-deficiency are thought to have an increased risk of cardiovascular disease. Sialic acid (SA) concentrations have been proposed as a marker of atherosclerotic disease probably related to an inflammatory response of the arterial wall. SA as a marker of cardiovascular disease in adult GH-deficiency and its relation to changes in fasting lipid profile and hormone concentrations have not yet been investigated. PATIENTS: We performed a randomised, double-blind placebo-controlled study in 18 patients with adult GH-deficiency before and after 3 months GH replacement therapy (0.036 U/kg/d; GH-treated group: 6 females, 3 males; age: 47.3 +/- 5.4 years., mean +/- SEM; placebo-group: 5 females, 4 males; mean age 50.2 +/- 4.7). In addition, SA concentrations were measured in 18 sex and age matched healthy control subjects. METHODS: Blood samples were obtained after an overnight fast. Serum SA, triglycerides and cholesterol were measured using enzymatic methods. Lipoprotein classes were separated by ultracentrifugation. Insulin and IGF-I were determined by radioimmunoassay, HbA1C was measured by anion exchange liquid chromatography. RESULTS: SA concentrations of the patients with adult GH-deficiency were not significantly different compared to the control group (GH-deficient group: 2.29 +/- 0.02 mmol/l, mean +/- SEM vs. control group: 2.09 +/- 0.13 mmol/l, P = 0.25). Before GH replacement therapy SA concentrations correlated positively with the patients age (r = 0.45; P < 0.04) and fasting insulin concentrations (r = 0.5; P < 0.03) but not with fasting lipid profile. GH replacement therapy significantly increased IGF-I (GH: + 27 +/- 2.6 vs. placebo: + 1.0 +/- 0.8 nmol/l, P < 0.001) and fasting insulin concentrations (GH: + 71.9 +/- 8.0 vs. placebo: + 19.6 +/- 22.6 pmol/l, P < 0.04) compared to placebo therapy. SA concentrations (GH: - 0.41 +/- 0.15 vs. placebo: - 0.01 +/- 0.12 mmol/l, P < 0.05), total cholesterol (GH: - 0.71 +/- 0.16 vs. placebo: 0.23 +/- 0.21 mmol/l, P < 0.003) and LDL-cholesterol ( 0.71 +/- 0.14 vs. placebo: - 0.12 +/- 0.21 mmol/l P < 0.04) significantly decreased after GH replacement therapy compared to placebo therapy. No significant correlation between changes in SA concentrations and changes in lipid profile were observed following GH replacement therapy. CONCLUSION: These results suggest that, firstly, GH replacement therapy may have a beneficial effect on the pathogenesis of atherosclerosis despite the increase in insulin concentrations, a surrogate marker of insulin resistance, secondly, the proposed beneficial effect of GH on the atherosclerotic process is likely to be multifactorial and cannot only be explained by changes in lipid profile and finally, SA might be a useful marker for the process of atherosclerotic disease in interventional studies. PMID- 10468988 TI - Classical pituitary apoplexy: clinical features, management and outcome. AB - OBJECTIVE: The term classical pituitary apoplexy describes a clinical syndrome characterized by sudden headache, vomiting, visual impairment and meningismus caused by the rapid enlargement of a pituitary adenoma usually due to haemorrhagic infarction of the tumour. Most published reports looking at the clinical features and management of pituitary apoplexy have not differentiated between patients with clinical and subclinical apoplexy, the latter diagnosed at surgery. Furthermore, little is reported on the clinical outcome, in particular visual and endocrinological, and the role of radiotherapy. The purpose of this study was to observe not only the clinical presentation but also the possible predisposing events, investigations, management, clinical outcome as well as the role of radiotherapy in patients presenting with classical pituitary apoplexy. PATIENTS AND DESIGN: In a retrospective analysis 1985-96, the medical records of 21 male and 14 female patients (mean age 49.8 years, range 30-74) with classical pituitary apoplexy were reviewed. This represents all patients seen with this condition over the stated period. MEASUREMENTS: In all patients, pre- and post- operative measurements were made of FT4, FT3, TSH, PRL, LH, FSH, cortisol (0900 h), GH, oestradiol (females) and testosterone (males). Pituitary imaging was by computerized tomography (CT) scan, magnetic resonance imaging (MRI) or both. RESULTS: Patients were followed for up to 11 years (mean 6.3 years: range 0.5 11). Headache (97%) was the commonest presenting symptom, followed by nausea (80%) and a reduction of visual fields (71%). Hypertension, defined as a systolic > 160 mmHg and/or a diastolic > 90 mmHg, was seen in 26% of patients. MRI correctly identified pituitary haemorrhage in 88% (n = 7), but CT scanning identified haemorrhage in only 21% (n = 6). By immunostaining criteria, null-cell adenomas were the most common tumour type (61%). Transsphenoidal surgery resulted in improvement in visual acuity in 86%. Complete restoration of visual acuity occurred in all patients operated on within 8 days but only in 46% of patients operated on after this time (9-34 days). Long-term steroid or thyroid hormone replacement was necessary in 58% and 45% of patients, respectively. Of the male patients, 43% required testosterone replacement, and long-term desmopressin therapy was required in 6%. Only two patients (6%) with tumour recurrence after transsphenoidal surgery for the initial apoplectic event, subsequently required radiotherapy. CONCLUSIONS: In classical pituitary apoplexy, headache is the commonest presenting symptom and hypertension may be an important predisposing factor. MRI is the imaging method of choice. Transsphenoidal surgery is safe and effective. It is indicated if there are associated abnormalities of visual acuity or visual fields because, when performed within 8 days, it resulted in significantly greater improvement in visual acuity and fields than if surgery was performed after this time. Radiotherapy is not indicated immediately as the risk of tumour recurrence is small, but careful follow-up initially with annual imaging is indicated in this group. PMID- 10468989 TI - Ontogeny of serum leptin concentrations in the human. AB - OBJECTIVE: Serum leptin concentrations reflect the fat mass of an individual. Fetal growth is rapid and it might be expected that major changes in circulating leptin concentrations in the fetus and neonate take place. We have studied the ontogeny of serum leptin concentrations in cord blood samples obtained by cordocentesis between 14 and 32 weeks of gestation and in samples obtained at term. PATIENTS: Cordocentesis samples from 10 appropriately grown for gestational age (AGA) and 10 intrauterine growth restricted (IUGR) fetuses. The results were compared with cord serum leptin concentrations obtained in 39 term healthy pregnancies. RESULTS: In the AGA and term pregnancies serum leptin concentrations changed little up until 38 weeks of gestation when there was an increase in concentration (Pre 38 weeks 3.5 microg/l (SEM 0.3); Post 38 weeks 5. 6 microg/l (SEM 0.7): Mann Whitney P = 0.03). Serum leptin concentrations were similar in the AGA and IUGR fetuses (AGA 5.9 microg/l (SEM 3.0); IUGR 4.2 microg/l (SEM 1.5): Mann-Whitney P = ns). Serum leptin was strongly related to birth weight (r = 0.58; P < 0.001) and birth length (r = 0.32; P = 0.05) in the term babies but not in the AGA or IUGR groups. CONCLUSION: Serum leptin concentrations in the fetus appear to be independent of gestational age and only rise towards the end of gestation. This late change probably reflects the changes in the accretion of body fat. There appears to be little difference in serum leptin concentrations between AGA and IUGR fetuses. PMID- 10468990 TI - The continuous 7-hour intravenous dexamethasone suppression test in the differential diagnosis of ACTH-dependent Cushing's syndrome. AB - OBJECTIVE: A recent report showing disappointingly low sensitivity and specificity for the oral high dose dexamethasone test in the differential diagnosis of Cushing's syndrome prompted us to re-evaluate the results obtained in our centre using the continuous 7-hour intravenous dexamethasone suppression test for this purpose. PATIENTS: 105 patients with ACTH-dependent Cushing's syndrome were included in this study; 78 with Cushing's disease, 8 with ectopic ACTH-secreting tumours and 19 were classified as 'of unknown aetiology'. RESULTS: In 74/78 (94.9%) of patients with Cushing's disease and in 3/8 (37.5%) patients with the ectopic ACTH syndrome, a plasma cortisol decrease > 190 nmol/l at 7 h as compared to baseline values was achieved in the continuous 7-hour intravenous dexamethasone suppression test. Using a plasma cortisol decrease > 190 nmol/l at 7 h as compared to baseline values as the cut-off value, the sensitivity and specificity of the continuous 7-hour intravenous dexamethasone suppression test for the diagnosis of Cushing's disease in patients with ACTH-dependent Cushing's syndrome were 94.9% and 62.5%, respectively. CONCLUSIONS: In patients with ACTH dependent Cushing's syndrome with a plasma cortisol decrease > 190 nmol/l at 7 h in the continuous 7-hour intravenous dexamethasone suppression test, additional localizing investigations such as bilateral simultaneous inferior petrosal sinus sampling and/or pentetreotide scintigraphy should be performed when no clearly discernible pituitary adenoma is observed on MRI studies. Patients with ACTH dependent Cushing's syndrome with a plasma cortisol decrease < 190 nmol/l at 7 h in the continuous 7-hour intravenous dexamethasone suppression test should also undergo bilateral simultaneous inferior petrosal sinus sampling and/or pentetreotide scintigraphy to demonstrate the presence of a nonpituitary source of ACTH overproduction. PMID- 10468991 TI - Leptin serum concentrations in healthy neonates within the first week of life: relation to insulin and growth hormone levels, skinfold thickness, body mass index and weight. AB - BACKGROUND AND AIMS: Leptin, the ob gene product, plays a key role in the regulation of body fat mass and weight in adult life. The mechanisms by which maternal and fetal/neonatal weight are regulated during human pregnancy and in early postnatal life are poorly understood. High leptin levels are observed in women during gestation and in cord blood at term. We have hypothesized that high leptin levels at term could represent an important feed-back indicator of nutrient supply. Subsequently, leptin could signal adipose tissue status during late gestation and during early neonatal life. SUBJECTS AND METHODS: 51 healthy newborns were studied. Clinical and auxological data (birth length, weight, and iliac, subscapular, biceps and triceps skinfold thickness) were recorded using a standardized data sheet. Venous cord blood was obtained immediately after birth in all neonates. Subsequently, capillary blood was obtained from the heel from some of the newborns when blood had to be obtained because of signs or symptoms of particular problems such as hypoglycaemia or hyperbilirubinaemia, at the following time points: two to four hours after birth in 51 infants, 56-79 h after birth in 47 infants and 99-128 h after birth in 23 of the newborns. The ratio between the sexes (girls/boys) was similar at all time points. The infants that were included in the study were subsequently found to be normal and healthy after analysis of the clinical and biochemical data. A specific ultrasensitive radioimmunoassay was used to measure leptin, while growth hormone and insulin were measured using commercially available immunoassays. RESULTS: Gestational age was 38-42 weeks, maternal age was 21-42 years. Birth weights ranged from 2480 to 4400 g. All newborns and mothers were subsequently found to be healthy. Leptin levels in venous cord blood was 0.16-6.80 microg/l, median 3. 47 microg/l and in capillary blood shortly after birth 0.26-7.03 microg/l, median 3.89 microg/l. 56 79 h after birth leptin levels had fallen dramatically, range 0.02-1.69 microg/l, median 0.26 microg/l, while 99-128 h after birth, leptin concentrations in capillary blood (0.05-2.61 microg/l, median 0.59 microg/l) had significantly increased when compared to the levels at 56-79 h (P < 0.001). There was a significant correlation between leptin levels in umbilical vein and birth weight of the neonates (r = 0.57, P < 0.03). Multistep regression analysis revealed that weight and skinfold thickness accounted for approximately 35-70% of the variation of leptin levels. Insulin and growth hormone, and glucose and bilirubin however, had no major impact on leptin levels. CONCLUSION: High leptin levels are present in cord blood at birth and in capillary blood shortly after birth. Since leptin levels in cord blood correlate with birth weight it is tempting to speculate that in the fetus as in later life leptin is signalling expansion of fat stores. Most importantly, we now report that leptin levels are high in the fetus but decline rapidly and dramatically after birth in healthy neonates. This may be important for the stimulation of feeding behaviour and the acquisition of energy homeostasis in the neonate. PMID- 10468992 TI - Sleep deprivation effects on the activity of the hypothalamic-pituitary-adrenal and growth axes: potential clinical implications. AB - OBJECTIVES: Although several studies have shown that sleep deprivation is associated with increased slow wave sleep during the recovery night, the effects of sleep deprivation on cortisol and growth hormone (GH) secretion the next day and recovery night have not been assessed systematically. We hypothesized that increased slow wave sleep postsleep deprivation is associated with decreased cortisol levels and that the enhanced GH secretion is driven by the decreased activity of the HPA axis. DESIGN AND SUBJECTS: After four consecutive nights in the Sleep Laboratory, 10 healthy young men were totally deprived of sleep during the fifth night, and then allowed to sleep again on nights six and seven. Twenty four hour blood sampling was performed serially every 30 minutes on the fourth day, immediately following the previous night of sleep and on the sixth day, immediately after sleep deprivation. MEASUREMENT: Eight-hour sleep laboratory recording, including electroencephologram, electro-oculogram and electromyogram. Plasma cortisol and GH levels using specific immunoassay techniques. RESULTS: Mean plasma and time-integrated (AUC) cortisol levels were lower during the postdeprivation nighttime period than on the fourth night (P < 0.05). Pulsatile analysis showed significant reduction of both the 24 h and daytime peak area (P < 0.05) and of the pulse amplitude (P < 0.01), but not of the pulse frequency. Also, the amount of time-integrated GH was significantly higher for the first 4 h of the postdeprivation night compared to the predeprivation night (P < 0.05). Cross-correlation analyses between the absolute values of the time-series of each hormone value and percentage of each sleep stage per half hour revealed that slow wave sleep was negatively correlated with cortisol and positively correlated with GH with slow wave sleep preceding the secretion of these hormones. In contrast, indices of sleep disturbance, i.e. wake and stage 1 sleep, were positively correlated with cortisol and negatively correlated with GH. CONCLUSION: We conclude that sleep deprivation results in a significant reduction of cortisol secretion the next day and this reduction appears to be, to a large extent, driven by the increase of slow wave sleep during the recovery night. We propose that reduction of CRH and cortisol secretion may be the mechanism through which sleep deprivation relieves depression temporarily. Furthermore, deep sleep has an inhibitory effect on the HPA axis while it enhances the activity of the GH axis. In contrast, sleep disturbance has a stimulatory effect on the HPA axis and a suppressive effect on the GH axis. These results are consistent with the observed hypocortisolism in idiopathic hypersomnia and HPA axis relative activation in chronic insomnia. Finally, our findings support previous hypotheses about the restitution and immunoenhancement role of slow wave (deep) sleep. PMID- 10468993 TI - Vitamin D insufficiency increases bone turnover markers and enhances bone loss at the hip in patients with established vertebral osteoporosis. AB - AIM: The aim of this study was to determine whether the presence of vitamin D insufficiency increases bone turnover and enhances bone loss by examining the relationship between bone turnover markers and Bone mineral density (BMD) in vitamin D insufficient and vitamin D sufficient patients, with established vertebral osteoporosis. SUBJECTS: 119 consecutive, active, community dwelling, elderly women were assessed over a 7-month period between the months of March to October. RESULTS: There was a significant correlation between parathyroid hormone (PTH) and 25 hydroxyvitamin D (25(OH)D), r = - 0. 42 (P < 0.01). The prevalence of vitamin D insufficiency was 26.9% (defined by a 25(OH)D >/= 6.1 microg/l and 4 years) growth hormone (GH) therapy on insulin sensitivity, glucose tolerance and lipid profile in non-GH deficient adolescents at completion of their growth. SUBJECTS: Thirty non-GH deficient (15 'idiopathic' short stature, 8 intrauterine growth retardation, 7 partial GH deficiency in childhood but normal on retesting) were recruited, median (range) age 16.9 years (15-20.3) prior to ceasing their GH therapy. Their median (range) duration of GH treatment was 7.9 years (4-11). Insulin sensitivity was also recorded in 10 normal controls with a median (range) age of 20.5 years (18.4-22.3). METHODS: Insulin sensitivity was assessed by a short insulin tolerance test in 18 patients on GH therapy and controls. It was repeated in 14 patients six months after stopping their GH therapy. A 3-h standard oral glucose tolerance test (OGTT) was performed in 19 patients on GH therapy, and repeated after 6 months off GH in 10 patients. Fasting lipids were also measured. RESULTS: Insulin sensitivity index was significantly lower in the patients on GH therapy than in the controls, (median (range)) 3.7%/min (1.2-5.3) and 5.3%/min (3.8-6.2), respectively. Six months after termination of GH therapy, insulin sensitivity increased significantly from 3.6%/min (1.2-5) to 4. 8%/min (2.8-5.6). Fasting plasma insulin decreased significantly off GH therapy from 10.1 to 3.6 mU/l. The area under the insulin curve during the OGTT was also significantly higher on GH therapy. Apart from one patient with impaired glucose tolerance on GH treatment, plasma glucose concentrations remained within the normal range. No lipid abnormalities were recorded. CONCLUSIONS: These data suggest that long-term GH therapy may cause insulin resistance in non GH deficient adolescents, but usually with neither impaired glucose tolerance nor hyperlipidaemia. PMID- 10469016 TI - Mutations in intron 3 of GH-1 gene associated with isolated GH deficiency type II in three Japanese families. AB - OBJECTIVE: Isolated GH deficiency (IGHD) type II is a disorder inherited in an autosomal dominant manner. Three mutations at the donor splice site of intron 3 of the GH-I gene have been identified in five families. In this report, we describe a novel mutation also at the donor splice site of intron 3 in patients with IGHD type II. PATIENTS: Five individuals diagnosed as IGHD: two sporadic cases and one family with three affected individuals (two siblings and their father). MEASUREMENT: Genomic DNA was extracted from peripheral mononuclear cells. All the exons and introns of the GH-I gene were amplified by polymerase chain reaction (PCR) and subjected to sequence analysis. RESULTS: A guanine to adenine transition at the fifth base of intron 3 (mutE), which has not been reported, was identified in the familial case but not in unaffected members of the family including the paternal grandparents. In the other two families with sporadic cases, a guanine to adenine transition at the first base of intron 3 (mutA) was identified in the affected subjects but not in other members of the families. CONCLUSION: MutE has not been previously reported and is the fourth mutation associated with IGHD type II. The guanine residue mutated in mutA was the second nucleotide of a CpG dinucleotide, which is regarded as a hot spot for mutations by a methylation-deamination mechanism. Since mutA has previously been identified in three type II IGHD kindreds belonging to different ethnic backgrounds, this appears to be the most frequent GH-I gene mutation in IGHD with a dominant inheritance. Because de novo mutations appeared to have occurred in all three families analyzed in the present study and the presence or absence of these mutations can easily be tested by PCR and restriction enzyme digestion, not only the familial cases but also sporadic cases with IGHD should be examined for a possible mutation at the donor splice site of intron 3 in the GH-1 gene. PMID- 10469018 TI - Hexarelin as a test of pituitary reserve in patients with pituitary disease. AB - BACKGROUND: The insulin tolerance test (ITT) is the reference standard for the diagnosis of cortisol and growth hormone (GH) deficiency, but problems have occurred in small children in inexperienced hands and it is contraindicated in patients with cardiac disease and epilepsy. Hexarelin is a growth hormone releasing peptide with GH-, ACTH/cortisol- and prolactin-releasing effects which involve both hypothalamic and direct pituitary mechanisms. We therefore investigated whether it could be used to test GH and ACTH/cortisol reserve in patients with pituitary disease. METHODS AND SUBJECTS: The changes in GH and cortisol in response to insulin-induced hypoglycaemia (intravenous human Actrapid 0.15 IU/kg) and hexarelin (2 microg/kg) in 19 patients with possible pituitary disease (5 males, mean age 39 years, range 21-70) were compared. The patients' responses during the hexarelin test were also compared to normal ranges of GH and cortisol responses established in healthy volunteers following hexarelin administration. RESULTS AND DISCUSSION: GH peak levels were significantly higher after hexarelin than after hypoglycaemia (mean +/- SEM; 67.1 +/- 16 vs. 26.9 +/- 6.8 mU/l respectively; P < 0. 001), while cortisol levels were significantly lower (420 +/- 34 vs. 605 +/- 50 nmol/l; P < 0.001). The peak responses of both hormones correlated significantly between the hexarelin and insulin-induced hypoglycaemia tests (r = 0.80, P < 0.001 for cortisol). Peak GH levels after hexarelin and ITT showed a significant positive correlation with IGF-I levels (r = 0.84 and r = 0.77, P < 0.001 for both). All patients with a subnormal GH response to hexarelin (<41.4 mU/l) had a peak GH response to ITT of <9 mU/l, and only one patient had a normal (although borderline) response to hexarelin with a subnormal GH response to the ITT. Although 17 of the 19 patients had corresponding cortisol responses to hexarelin and the ITT test (either failing or passing both), two patients had normal cortisol responses to hexarelin but subnormal responses to the ITT. A peak serum cortisol level following hypoglycaemia of >580 nmol/l is indicative of normal cortisol reserve, as established in patients undergoing surgery; only five of the normal volunteers and one of the thirteen patients with a normal ACTH/cortisol reserve on ITT had a peak cortisol >580 nmol/l in response to hexarelin. CONCLUSION: Adult patients who have a subnormal peak GH response to hexarelin are likely to be GH deficient on an insulin tolerance test. However, our data suggest that the hexarelin test is not a useful test of ACTH/cortisol reserve. The hexarelin test could be a useful first/screening test to diagnose adult GH deficiency, particularly in patients in whom an insulin tolerance test is contraindicated or who are already ACTH deficient and in whom the GH reserve alone is of interest. PMID- 10469017 TI - The prevalence of thyroid dysfunction in a population with borderline iodine deficiency. AB - OBJECTIVE: We lack information on the influence of borderline iodine deficiency on the occurrence of thyroid dysfunction. Iodine deficiency has been reported to facilitate the development of toxic nodular goitre, whereas a high iodine intake may increase the prevalence of autoimmune hypothyroidism. SUBJECTS AND METHODS: In a cross-sectional study of a random sample of the general population in our region with borderline iodine deficiency 2656 (65%) of 4073 men and women aged 41 to 71 years participated. Records were made of previous thyroidal illness. Blood samples were drawn for thyroid parameters and TPO Ab values. Iodine and creatinine was assessed in casual urine samples. RESULTS: Previous or present hyperthyroidism was reported by 1.4% of the participants whereas 0.6% had unknown biochemical hyperthyroidism. All cases of undiagnosed hyperthyroidism were among women. Previously diagnosed and treated hypothyroidism was reported by 1.0% and undiagnosed hypothyroidism was found in 0.4%. Subclinical hyperthyroidism was found in 1.3% and subclinical hypothyroidism in 0.7%. TPO Ab titres >200 kU/l were found in 16.9% of the women and 6.6% of the men, and 83% of participants with TSH >5 mU/l had TPO Ab titres >200 kU/l. Participants with TPO Ab titres between 100 and 200 kU/l had no increased frequency of thyroid dysfunction. The median iodine excretion rate was estimated as 103 microg/day. Serum TSH values were higher in women than in men and showed higher dispersion in women as well as in old age. Serum free T3 was found to be higher in women than in men and increased with age. Serum free T4 showed no sex difference but values increased with increasing age. CONCLUSION: In our region with borderline iodine deficiency more than 5% of the general population has clinical or subclinical thyroid dysfunction. We found a relatively high prevalence of hyperthyroidism, especially previously undiagnosed disease, but a low prevalence of hypothyroidism as would be expected in an area of iodine deficiency. Hypothyroidism was related to TPO Ab titres of >200 kU/l. Thyroid hormone levels varied with age and sex. PMID- 10469019 TI - Pituitary apoplexy following metastasis of bronchogenic adenocarcinoma to a prolactinoma. AB - A 42-year-old house wife presented with worsening headaches over 6 months in the absence of visual symptoms or symptoms suggestive of focal neurology. She was a life-long smoker. Systems review was unremarkable apart from secondary amenorrhoea and galactorrhoea of 6 months duration. Her serum prolactin was found to be 620 mU/l (60-400), FT4 12.6 nmol/l (9.8-23.1), TSH 1.38 mU/l (0.35-5.5), oestradiol < 73 pmol/l, LH and FSH of 4.4 and 12.6 mIU/l, respectively. She was on bromocriptine. A presumptive diagnosis of pneumonia, based on pyrexia and CXR findings, was made and she was started on IV antibiotics. Two days later she developed meningism and deterioration of conscious level. (Lumbar puncture results: no organisms, 312 neutrophils and 164 lymphocytes). CT scan revealed a 2.5-cm pituitary adenoma, with suprasellar extension. A repeat hormonal profile revealed FSH 1.4, LH < 0.3 mU/l, oestradiol < 73 pmol/l, prolactin 488 mU/l (60 400), and low random cortisol at 29 nmol/l. T1-weighted MRI revealed a large pituitary mass with evidence of haemorrhage. The patient subsequently underwent a transsphenoidal exploration with resection of the pituitary lesion. Whilst awaiting the histopathology results, CT of chest revealed a 1. 5-cm diameter rounded well defined density in the right lower lobe associated with hilar, pre- and right para-tracheal lymphadenopathy. The histopathology of the pituitary lesion, obtained piecemeal, revealed fragments of fibrous tissue infiltrated by sheets of acidophilic prolactin-positive cells, in keeping with a prolactinoma. In addition, other fragments with blood clot included highly atypical epithelial cells with mitotic figures. These were negative for prolactin but showed HMFG-and CEA-positivity, excluding them from a pituitary lineage. Transbronchial biopsy revealed moderately differentiated adenocarcinoma, with evidence of lymphatic spread. The overall conclusion was of bronchogenic adenocarcinoma, metastasizing to a prolactinoma and complicated by apoplexy. PMID- 10469020 TI - Beta2-adrenoceptor polymorphisms and asthma. PMID- 10469021 TI - Cloning T cells from sites of allergic inflammation. PMID- 10469022 TI - Fruit allergy: plant defence proteins as novel potential panallergens. PMID- 10469023 TI - Anisakiasis: immunity, allergy or both? Lessons on the natural role of immunoglobulin E from a nematode infestation. PMID- 10469024 TI - Interleukin-10: an anti-inflammatory cytokine with therapeutic potential. PMID- 10469025 TI - Eosinophil cationic protein (ECP): molecular and biological properties and the use of ECP as a marker of eosinophil activation in disease. PMID- 10469026 TI - Mechanisms of particulate matter toxicity. PMID- 10469027 TI - Polymorphisms in the beta2-adrenoreceptor gene are associated with decreased airway responsiveness. AB - BACKGROUND: There are a number of candidate genes thought to play a role in the development of asthma. Polymorphisms at amino acid positions 16 (arginine to glycine) and 27 (glutamine to glutamic acid) of the beta2-adrenoreceptor (B2AR) gene are known to be functionally relevant and have been associated with more severe forms of asthma, nocturnal asthma and decreased airway responsiveness in asthmatic subjects. OBJECTIVE: To determine if these polymorphisms contribute to the development of asthma by investigating the associations between the polymorphisms at amino acid positions 16 and 27 of the B2AR gene and asthma related parameters in a large, phenotypically well-characterized population which was unselected for asthma. METHODS: Subjects (n = 332) were characterized using physiological assessments, immuno-logical data and information obtained from questionnaire. PCR was used to generate a 229 base pair fragment spanning the mutations of interest. Genotype was determined using allele-specific oligonucleotide hybridization and confirmed in 10% of the samples by direct sequencing. Multivariate analysis of the association between genotype and phenotype was then undertaken. RESULTS: Homozygotes for Glu27 were significantly less responsive to histamine than Gln27 homozygotes. In addition, Arg16 homozygotes were more likely to 'wheeze during a cold', in comparison with Gly16 homozygotes. However, there was no association between either polymorphism and physician-diagnosed asthma, eczema, skin reactivity to common allergens or total and specific serum IgE levels. The two polymorphisms were found to be in significant linkage disequilibrium. CONCLUSION: The polymorphism at position 27 was associated with decreased airway responsiveness in the study population and the polymorphism at position 16 was associated with increased wheeze during respiratory infection, but neither was associated with physician-diagnosed asthma or any of the other variables considered. PMID- 10469028 TI - Association between - 308 tumour necrosis factor promoter polymorphism and bronchial hyperreactivity in asthma. AB - BACKGROUND: Tumour necrosis factor (TNF) is a pivotal cytokine in the inflammation underlying asthma. The TNF gene is located in the polymorphic HLA class 3 region on chromosome 6p. Several polymorphisms in this region have been described and associated with alteration of TNF secretion in vitro. OBJECTIVE: In this study we tested the hypothesis that two such polymorphisms, lymphotoxin alpha NcoI B*1 and -308 TNF2 may be components of the genetic predisposition to asthma. METHODS: Five hundred and fifty-six random individuals were studied, comprising approximately equal numbers of asthmatic subjects, with or without atopy, and a nonatopic nonasthmatic control group. In addition, 355 subjects (172 asthmatics) from 60 multiplex families were typed at the LTalpha NcoI locus. RESULTS: There was an association between allele two of the -308 TNF polymorphism and bronchial hyperreactivity (OR 2.12, 95% CI 1.04-4.32, P = 0.036). However, there was no association with LTalpha NcoI alleles. To determine whether this was influenced by linkage disequilibrium within the MHC, 91 subjects with bronchial hyperreactivity and 85 control subjects were typed for class 2 and 3 alleles. Following identification of the extended TNF2 haplotype, we found no independent association of these alleles with BHR. CONCLUSIONS: We conclude that the -308 TNF2 promoter polymorphism may form a component of the genetic predisposition to BHR in asthma. PMID- 10469029 TI - Elevated levels of soluble TNF receptors in bronchoalveolar lavage fluid in extrinsic allergic alveolitis. AB - BACKGROUND: Soluble TNF receptors (sTNFR1 and sTNFR2) are inhibitors of TNF and can block TNF bioactivity. TNF plays an important role in the development of extrinsic allergic alveolitis (EAA). OBJECTIVE: To evaluate whether sTNFR1 and sTNFR2 are locally increased in EAA. METHODS: We measured sTNFR1 and sTNFR2 in bronchoalveolar lavage fluid (BALF) and serum from nine EAA patients and 11 control subjects using an ELISA method. RESULTS: BALF sTNFR1 and sTNFR2 levels were 0.24+/- 0.04 ng/mL and 0.59+/-0.16 ng/mL in EAA patients, and thus significantly elevated in comparison with the controls (0.13+/-0.02 ng/mL and 0.08+/-0.04 ng/mL, both P<0.05). Serum sTNFR levels were not significantly different between the two groups. Both sTNFR1 and sTNFR2 concentrations in BALF correlated significantly with the lymphocyte percentage of BALF (r = 0.57 and 0.81, respectively). CONCLUSION: The two alveolar sTNFRs, particularly sTNFR2, may be involved in the pathogenesis of EAA as counter-regulators of TNF. PMID- 10469030 TI - Cytokine production and mRNA expression by conjunctival T-cell lines in chronic allergic eye disease. AB - BACKGROUND: Activated CD4+ T cells, mast cells and eosinophils are the main cytokine-producing cell-types infiltrating the conjunctiva during chronic allergic eye diseases. Interactions between these cells are thought to play an important immunopathogenic role in these disorders (giant papillary conjunctivitis; vernal keratoconjunctivitis; atopic keratoconjunctivitis). OBJECTIVE: The objective was to compare the cytokine profiles of conjunctival T cell lines from patients with different forms of chronic allergic eye disease. METHODS: T cells were isolated from conjunctival biopsies and non-specifically expanded into lines. The lines were immunophenotyped by flow cytometry. Cytokine production was quantified by immunoassays and more sensitive molecular techniques were used to investigate cytokine mRNA expression to identify the presence of interleukin (IL) -2, IL-4 and interferon (IFN) -gamma transcripts. RESULTS: Following four to six rounds of stimulation, the conjunctival T-cell populations were CD3+ (> 93%), with variable levels of CD4 and CD8 expression. All were HLA DR+ (> 80%) with some HLA-DQ expression. Conjunctival T-cell lines from atopic keratoconjunctivitis produced selective increases in IFN-gamma, IL-10 and IL-13 (P<0.01), those from vernal keratoconjunctivitis produced increased IL-5 (P<0.01) whereas T-cell lines from giant papillary conjunctivitis produced only low levels of cytokines. IL-4 was only detected at the mRNA level and was expressed in four out of five T-cell lines in the vernal keratoconjunctivitis group. In contrast there was moderate to strong expression of IFN-gamma in five out of six T-cell lines in atopic keratoconjunctivitis. CONCLUSION: Different patterns of T-cell cytokine profiles were observed for each disease, with low-level, non-polarized cytokine production in giant papillary conjunctivitis, a TH2-like profile in vernal keratoconjunctivitis and a shift towards a TH1-like profile in atopic keratoconjunctivitis. PMID- 10469031 TI - Regulation of T-helper cell responses to inhalant allergen during early childhood. AB - BACKGROUND: Recent evidence suggests that preschool children manifest patterns of allergen-specific skin prick test (SPT) reactivity and in vitro T-cell cytokine production which are similar to that of either atopic or nonatopic adults. However, published studies on this age group involve small sample sizes and a restricted number of cytokines, usually in response to polyclonal stimuli. OBJECTIVE: To elucidate the relationship between in vivo and in vitro immune responses to a major inhalant allergen house dust mite (HDM) in preschoolers. METHODS: Peripheral blood mononuclear cells (PBMCs) from matched groups of HDM SPT+ and SPT- 6-year-olds (n = 30 and 29, respectively) tested for PBMC responses to HDM, and cytokine production measured at both the protein and mRNA levels. Immunoglobulin (Ig) E and IgG subclass antibody titres were determined in serum. Interrelationships between in vitro and in vivo HDM responses were examined via multivariate analyses. RESULTS: SPT reactivity to HDM was associated with in vitro production by putative T cells of interleukin (IL) -4, IL-5, IL-9, IL-10, IL-13 and low level IFNgamma, and with production in vivo of IgE and (all) IgG subclass antibodies; HDM responses in the SPT- group were restricted mainly to IL 10 and IFNgamma and very low levels of IL-4; IL-6 production from non-T-cell sources was common. The cytokine most associated with positive SPT responses was IL-9; SPT weal diameter correlated positively with IL-4, IL-5 and IL-13 and negatively with IL-10. CONCLUSION: Detailed analysis of cytokine responses in this very young age group have the potential to uncover subtle relationships between in vivo and in vitro allergen reactivity which may be less clear in adults, in whom T-cell response patterns are modified via chronic stimulation. The present findings which suggest potentially important roles for IL-9 and IL-10 in the early phase of allergic disease, may be one such example. PMID- 10469032 TI - Detection of inhaled Der p 1. AB - BACKGROUND: Measurement of personal exposure to Der p 1 aeroallergen has previously been limited by the low quantity of material collected by sampling systems and the assay sensitivity. This has meant that exposure could only be detected if long sampling periods were used or reservoir dust was artificially disturbed. We have developed a sampling method to sample true personal exposure and combined it with a novel method which is sensitive enough to measure allergen exposure over shorter time frames. OBJECTIVE: To describe normal domestic exposure to dust mite allergen during a range of activities in houses in Sydney, Australia. METHODS: Inhaled particles containing mite allergen Der p 1 were collected using a nasal air sampler which impacts particles (> approximately 5 microm) onto a protein-binding membrane coated with a thin, porous, adhesive film. The allergen is bound to the membrane in the immediate vicinity of the particle and detected by immunostaining with monoclonal antibodies specific for Der p 1. In addition, samples were collected using a standard Institute of Occupational Medicine (IOM) personal air sampler and the amount of eluted Der p 1 was assayed by ELISA. RESULTS: The median number (range) of inhaled particles containing Der p 1 collected in each 10-min sampling period was: dust raising 5 (2-10); lying in bed, 0 (0-2); sitting on the bed, 1 (0-2); walking around the bedroom, 0 (0-2). This represented 0-5.1% of all particles captured. The Der p 1 concentration of floor and bed dust was 19.4 and 55.1 microg/g, respectively. The standard IOM personal sampler and ELISA were unable to detect Der p 1 for any of the activities performed. CONCLUSIONS: We were able to count individual allergen carrying particles inhaled over short time periods, during different domestic exposure situations. This will offer new insight into several aspects of personal allergen exposure. PMID- 10469033 TI - Peels of Rosaceae fruits have a higher allergenicity than pulps. AB - BACKGROUND: It is not uncommon that patients allergic to fruits such as apple, pear, and peach, refer adverse reactions after the ingestion of the whole fruit, but subsequently tolerate the pulp. OBJECTIVE: This study aimed to compare the allergenicity of peels and pulps of apple, peach, and pear in 33 patients allergic to these fruits. METHODS: Clinical reactivity to the ingestion of whole fruit (peel + pulp) and pulp was established by medical history. Peels and pulps were tested separately in skin prick tests (SPTs), histamine release tests (HRTs) and RASTs. Cross-allergenicity between peel and pulp of apple and peach was studied by RAST inhibition. RESULTS: Adverse reactions appeared more frequently and were more severe when the whole fruit was eaten. More than 40% of patients allergic to apple and pear tolerated the ingestion of the pulp of these fruits, and reactions were only elicited by the intake of the whole fruit. Peels induced higher SPTs, HRTs and RASTs than pulps. An important cross-allergenicity was found between the peel and pulp of apple and peach, although the amount of the shared allergenic epitopes seemed to be higher in peels. CONCLUSION: Our results suggest that peels of Rosaceae fruits such as apple, peach, and pear, have a higher allergenicity than pulps, which is clinically relevant. This aspect should be considered in the evaluation of patients allergic to Rosaceae fruits, and in the production of diagnostic materials. PMID- 10469034 TI - Composition and stability of allergenic extracts made from gamma-irradiated rye grass (Lolium perenne) pollen. AB - BACKGROUND: Phenol is commonly added to allergenic extracts as a bacteriostatic agent, but it is poisonous and also detrimental to proteins, which accelerates extract degradation. Sterilization by gamma-irradiation of the source material could be an alternative to the use of phenol. OBJECTIVE: To analyse the potential effects of gamma-irradiation of pollen on the composition, potency, and stability of the resulting extract, and compare them with those of phenol. METHODS: Ryegrass (Lolium perenne) pollen was sterilized by gamma-irradiation at a dose of 25 kGy. Extracts prepared from the irradiated pollen were then compared by electrophoresis techniques and RAST inhibition to extracts, without or with 0.5% phenol, from nonirradiated pollen. In addition, proteolytic activity was compared in extracts from irradiated and nonirradiated pollen. To evaluate the stability of extracts on storage, they were analysed after forced degradation for up to 7 days at 37 degrees C. RESULTS: When fresh extracts were analysed, there were no noticeable differences between the three types, as judged by immunoblotting and RAST inhibition experiments. However, on storage, extracts from irradiated pollen appeared to be superior to extracts from nonirradiated pollen, as some proteins were more stable in the former. This could be related to the lower proteolytic activity we have also observed in extracts from irradiated pollen. In contrast, extracts containing phenol degraded much faster, as proven by all our methods of investigation. CONCLUSION: Gamma-irradiation of pollen did not influence the IgE binding capacity of the resulting extracts, but did yield extracts with somewhat improved stability, probably by reducing the proteolytic activity. It may be concluded that gamma-irradiation of the source material represents a good alternative to the use of phenol for the preparation of allergenic extracts. PMID- 10469035 TI - Specific immunoglobulin E to peanut, hazelnut and brazil nut in 731 patients: similar patterns found at all ages. AB - BACKGROUND: Previous studies have reported reactions to an increasing range of nuts as patients with nut allergy grow older. Most patients with symptoms suggesting nut allergy have specific IgE to more than one nut. Furthermore, fatal reactions have followed eating nuts different from any causing the deceased's previous reactions. OBJECTIVE: To explore the pattern of specific IgE to three distantly related nuts in patients of all ages with nut allergy. METHODS: This study includes all patients referred to our laboratory for nut allergy testing from January 1994 to August 1998 who were tested for peanut, hazelnut and brazil nut, and had specific IgE to at least one of these nuts. All tests were performed using the Pharmacia Unicap system. RESULTS: Seven hundred and thirty-one patients (age 7 months to 65 years, median 6.6 years) had specific IgE >0.35 kU(A)/L to at least one of these three nuts: 282 had IgE to one nut, 130 to two nuts, and 319 to all three nuts. When analysed by gender and age quartile, very similar patterns were found in all subgroups though significant age trends and age interactions were found for IgE to individual nuts and combinations of nuts. CONCLUSIONS: The probability of a patient with nut allergy having specific IgE to a particular combination of peanut, hazelnut and brazil nut is similar, whatever their age or sex. The apparent increase in multiple nut reactivity with increasing age may therefore be due to exposure of previously unchallenged sensitivity. The frequency of multiple-nut specificity is sufficiently high that patients should always be tested for allergy to a range on nuts if they have a history of reacting to any nut. PMID- 10469036 TI - Usefulness of early serial measurement of specific and total immunoglobulin E in the diagnosis of gastro-allergic anisakiasis. AB - BACKGROUND: Sensitization to Anisakis simplex (A. simplex) has been documented to produce severe allergic reactions following ingestion of mainly raw or under cooked parasitized fish. False positive skin prick tests (SPT) or specific IgE against this nematode and cross-reactivity restricts diagnosis. Gastric anisakiasis and gastro-allergic anisakiasis occur if fish is parasitized by live A. simplex OBJECTIVE: To investigate if serial serological analysis could be useful in the diagnosis of acute parasitation by this nematode. METHODS: We included 41 patients who experienced an allergic reaction and/or abdominal symptoms after ingestion of raw or undercooked fish and displayed specific IgE against A. simplex. Total and specific IgE were determined two times: in the 24-h period after onset of clinical symptoms and after 1 month. SPTs were performed against A. simplex and implicated fish. A fibre optic gastroscopy was performed in 22 patients. RESULTS: Median total IgE was 80.0 (Interquartile range [IQR] 41.5-186.5) kU/L in the first evaluation and 247.0 (IQR 96.5-649.5) kU/L after 1 month. Median specific IgE against A. simplex was 11.4 (IQR 7.1-33.5) kU/L in the first 24 h and 36.8 (IQR 19.5-79.5) kU/L after 1 month. A rise of total IgE was observed in 35 of 41 patients (P<0.00001) and a rise in specific IgE against A. simplex in 37 of 41 patients (P<0.00001). Mean percentage increment was 392% (215 571%; 95% confidence interval [CI]) for total IgE and 339% (177-502%; 95% CI) for specific IgE. In nine of 22 gastroscopic examinations at least one larva, identified as A. simplex, could be detected by our microbiology service. In this group (n = 9) a rise of total and specific IgE was detected in eight patients (89%) (P = 0.02). CONCLUSIONS: We consider a rise of total and specific IgE in the first month after an allergic reaction as a useful tool in the diagnosis of gastro-allergic anisakiasis (together with patient's history), even if the parasite cannot be seen with fibre optic gastroscopy. The important rise of total and specific IgE against A. simplex can be considered as a reaction induced by the live parasitizing larva in the context of a polyclonal immunological stimulation. PMID- 10469038 TI - Exhaled monoxides in asymptomatic atopic subjects. AB - BACKGROUND: Atopy is a genetically determined condition and some atopic people develop airway hyperresponsiveness and sometimes asthma later in life. Since airway inflammation may be present before the onset of clinical symptoms of asthma, early and noninvasive detection of inflammation would be useful in atopic subjects. Mediators produced by activated inflammatory cells may lead to induction of inducible nitric oxide synthase producing nitric oxide (NO) and inducible heme oxygenase releasing carbon monoxide (CO) in the airways. Both monoxides are present in exhaled air and their levels are elevated in asthma reflecting airway inflammation. OBJECTIVE: We have measured exhaled NO and CO levels in atopic and nonatopic healthy non-smoking subjects to determine whether inflammation is present in the airways. METHODS: Exhaled NO was measured by a chemiluminescence analyser and exhaled CO electrochemically and NO in asymptomatic atopic and age-matched nonatopic normal subjects. RESULTS: Exhaled NO and CO levels were both significantly elevated in 15 atopic subjects compared with 40 nonatopic individuals (means +/- SEM: 18.3+/-3.0 p.p.b. vs. 6.3+/-0.3 p.p.b., P< 0.0001 and 4.7+/-0.3 p.p.m. vs 2.8+/-0.2 p.p.m., P = 0.0005, respectively). CONCLUSION: Increase in exhaled monoxide levels may be an early and noninvasive marker of airway inflammation in asymptomatic atopic subjects. PMID- 10469037 TI - Pollinosis to Ricinus communis (castor bean): an aerobiological, clinical and immunochemical study. AB - BACKGROUND: Ricinus communis (castor bean) is a species included into the Euphorbiaceae family, common to all the warm regions of the world. Although the allergenicity of its seed is well known, references are scarce regarding the role played by its pollen as a pneumo-allergen. OBJECTIVES: To carry out an aerobiological study of this pollen in the Malaga area (southern Spain); describe the physicochemical characteristics of its most relevant allergens; and to demonstrate the existence of patients with respiratory allergy due to this pollen. METHODS: A Burkard spore trap was used for the aerobiological study from 1992 to 1996. Skin prick tests with castor bean pollen extract were performed to 1946 patients with rhinitis and/or asthma. Specific IgE levels were measured in castor bean-positive SPT patient sera. Immunochemical characterization of the most relevant allergens was performed using electrophoretic techniques. In vitro cross-reactivity studies using positive patient sera were carried out. Nasal challenge tests were done in 32 subjects randomly selected from the sensitized patient group. RESULTS: Castor bean is a perennial pollen with total annual pollen levels never exceeding 1%. One hundred and eighteen (7.7%) patients showed positive prick test (74 rhinitis, 36 rhinitis and asthma, eight asthma). Nine were monosensitized. Specific IgE levels were > or =0.35 PRU/mL in 39 (33%) of patient sera. Nasal challenge test: 10 subjects presented non-specific nasal hyperactivity, 15 were positive and seven negative. The molecular masses and isoelectric points of the main IgE-binding proteins, ranged from approximately 67 15.5/14.5 kDa and approximately 4.5-5.5, respectively. Profilin of the extract was purified by poli-L-proline-Sepharose chromatography and it appeared as one of the most frequent allergens. CONCLUSION: Castor bean pollen is an allergen which causes respiratory (mainly nasal) symptoms. PMID- 10469039 TI - Specific inhibition of TH2-type cytokine production from human peripheral T cells by terfenadine in vitro. AB - BACKGROUND: Cytokine imbalance is thought to be one of the causes for allergic diseases. The effect of anti-allergic drugs on cytokine production from T cells should be examined in a convenient way. OBJECTIVES: To study the in vitro effect of terfenadine, a prototype non-sedating H1 receptor antagonist, on cytokine production from activated T cells. METHODS: T cells were cultured in the presence of terfenadine on anti-CD3 mAb and anti-CD26 mAb-coated wells, anti-CD3 mAb and anti-CD28 mAb-coated wells, and anti-CD3 mAb wells with PMA. T-cell proliferation, along with the concentrations of interleukin (IL) -2, interferon (IFN) -gamma, IL-4, and IL-5 were measured. RESULTS: Terfenadine inhibited T-cell proliferation and IL-4 and IL-5 production under each costimulatory condition tested, whereas it had no effect on IL-2 and IFN-gamma production. CONCLUSIONS: These results indicate that terfenadine has a specific inhibitory effect on TH2 type cytokine production induced by several ways of costimulatory activation. PMID- 10469040 TI - Protective effect of oral terfenadine and not inhaled ipratropium on adenosine 5' monophosphate-induced bronchoconstriction in patients with COPD. AB - BACKGROUND: Inhalation of adenosine 5'-monophosphate (AMP) causes bronchoconstriction in patients with asthma and in many patients with chronic obstructive pulmonary disease (COPD). In asthma, AMP-induced bronchoconstriction has been shown to be determined mainly by release of mast cell mediators, and possibly by vagal nerve stimulation, since oral terfenadine (H1-receptor antagonist) and inhaled ipratropium bromide (muscarinic receptor antagonist) both increase PC20AMP. OBJECTIVE: To investigate the mechanism of AMP-induced bronchoconstriction in COPD. METHODS: We performed a randomized, double-blind, placebo-controlled, crossover trial. Forty-four nonatopic hyperresponsive smokers with COPD (mean age +/- SD: 60+/-7 years, FEV1 61+/-12% of predicted and FEV1/VC 51+/-8%, geometric mean [GM] PC20methacholine 0.62 mg/mL and GM PC20AMP 6.77 mg/mL) participated. PC20methacholine and PC20AMP were assessed on 3 days. Before the challenges they used either 180 mg of oral terfenadine, 120 microg of inhaled ipratropium bromide, or placebo. RESULTS: GM PC20AMP was 5.44 mg/mL after placebo, increasing with 0.9 doubling concentration (P<0.0001) after terfenadine and decreasing 0.3 doubling concentration after ipratropium bromide (NS). GM PC20methacholine was 0.75 mg/mL after placebo, increasing 0.4 doubling concentration after terfenadine (NS) and 3 doubling concentrations after ipratropium bromide (P<0.0001). CONCLUSION: These findings indicate that histamine release is important in the pathophysiology of AMP-induced bronchoconstriction in smokers with COPD, whereas vagal nerve stimulation does not play a role. Therefore, PC20AMP may be a valuable tool in evaluation of treatments which affect airway histamine release. PMID- 10469041 TI - Regeneration of autotransplanted splenic fragments: basic immunological and clinical relevance. PMID- 10469042 TI - Two polymorphisms of the tumour necrosis factor gene do not influence survival in pancreatic cancer. AB - Polymorphisms of the tumour necrosis factor (TNF) gene have been related to TNF production and outcome in a variety of inflammatory and malignant diseases. Proinflammatory cytokines and the inflammatory state appear to affect outcome in pancreatic cancer. Thus, the present study examined the TNFB and TNF-308 polymorphisms for their relationship to the inflammatory state and survival in pancreatic cancer. Sixty-four patients with advanced pancreatic cancer and 101 healthy subjects were genotyped for each polymorphism. Serum concentrations of the two TNF receptors and C-reactive protein (CRP) were measured in 45 of the cancer patients with no evidence of infection or jaundice, 1 month after surgical intervention. There was no difference in distribution of genotypes between the patient and control groups. There was no association between any genotype and concentrations of any of the measured inflammatory mediators. While those with an elevated CRP concentration had significantly poorer survival, there was no association between either TNF genotype and survival. This study found no association between TNF genotype and the inflammatory state or survival in advanced pancreatic cancer. Other cytokines may be more important than TNF in determining the inflammatory state and disease progress in pancreatic cancer. PMID- 10469043 TI - Effect of shear stress and a stable prostaglandin I2 analogue on adhesive interactions of colon cancer cells and endothelial cells. AB - In the process of cancer metastasis, adhesion between cancer cells and endothelial cells is an important early step. In the present study, the effects of shear stress and the adhesion molecules responsible for cancer cell interactions with endothelial cells were investigated in a system similar to in vivo microcirculation. The effect of prostaglandin I2 (PGI2) also was determined. Human colon cancer cell line Colo 201 and human umbilical vein endothelial cells (HUVEC) were used. After HUVEC on a glass slide were incubated with IL-1beta for 4 h, cancer cells in suspension were perfused on HUVEC at wall shear stresses of 5-40 microN/cm2. Experiments were videotaped, and the number of adherent cells were counted. Additionally, the effects of anti-sialyl Lewis a (SLea) MoAb, anti E-selectin MoAb, and a PGI2 analogue were investigated. Expression of adhesion molecules on cancer cells and HUVEC was assessed using flow cytometry and enzyme immunoassay, respectively. Few cancer cells adhered to HUVEC without IL-1beta; however, many cancer cells adhered to IL-1beta-stimulated HUVEC at low shear stress (5-20 microN/cm2). Cancer cells did not migrate beneath HUVEC. The increased adhesion was inhibited by anti-E-selectin MoAb, anti-SLea MoAb, and a PGI2 analogue. In addition, the PGI2 analogue decreased the surface expression of SLea on Colo 201 cells. These results suggest that Colo 201 cells adhere to IL 1beta-stimulated endothelial cells via SLea and E-selectin under low flow conditions; PGI2 analogues may protect against metastasis by inhibiting cancer cell-endothelial cell interactions. PMID- 10469044 TI - Enhancement of lectin pathway haemolysis by immunoglobulins. AB - We recently reported that indicator sheep erythrocytes (E) coated with mannan and sensitized with mannan-binding lectin (MBL) (E-M-MBL) are lysed by human serum in the absence of calcium via the lectin pathway of complement activation by a process which requires alternative pathway amplification and is associated with increased binding of and control by complement regulatory proteins C4 bp and factor H. In the present study, we investigated the effect of immunoglobulin (Ig) on this haemolysis. Co-sensitization of indicator E with anti-E haemolysin led to threefold enhancement of lectin pathway haemolysis in the absence of calcium, associated with increased binding of C3 and C5. Lysis was enhanced approximately twofold when E-M-MBL were chemically or immunologically coated with IgM or IgA, and fourfold when coated with IgG, prior to lysis in human serum-Mg ethyleneglycol tetraacetic acid. The presence of haemolysin did not reduce the binding or inhibitory activity of C4 bp, and the enhancing activity of haemolysin was retained in serum depleted of C4 bp. By contrast, binding of factor H was greatly reduced in the presence of haemolysin, which had no enhancing effect in serum depleted of factor H. These experiments demonstrate the ability of IgG, IgM and IgA to enhance lectin pathway cytolysis, and that this enhancement occurs by neutralization of the inhibitory activity of factor H. Immunoglobulin enhancement of lectin pathway cytolysis represents another interaction between the innate and adaptive systems of immunity. PMID- 10469045 TI - Mechanism of complement-dependent haemolysis via the lectin pathway: role of the complement regulatory proteins. AB - Mannan-binding lectin (MBL) is an acute phase protein which activates the classical complement pathway at the level of C4 and C2 via two novel serine proteases homologous to C1r and C1s. We recently reported that haemolysis via this lectin pathway requires alternative pathway amplification. The present experiments sought to establish the basis for this requirement, and hence focused on the activity and regulation of the C3 convertases. Complement activation was normalized between the lectin and classical pathways such that identical amounts of bound C4 and of haemolytically active C4,2 sites were present on the indicator cells. Under these conditions, there was markedly less haemolysis, associated with markedly less C3 and C5 deposited, via the lectin pathway than via the classical pathway, particularly when alternative pathway recruitment was blocked by depletion of factor D. Lectin pathway activation was associated with enhanced binding in the presence of MBL of complement control proteins C4bp and factor H to C4b and C3b, respectively, with decreased stability of the C3-converting enzyme C4b,2a attributable to C4bp. Immunodepletion of C4bp and/or factor H increased lectin pathway haemolysis and allowed lysis to occur in absence of the alternative pathway. Thus, the lectin pathway of humans is particularly susceptible to the regulatory effects of C4bp and factor H, due at least in part to MBL enhancement of C4bp binding to C4b and factor H binding to C3b. PMID- 10469046 TI - Galactosylation of serum IgG and autoantibodies in murine models of autoimmune haemolytic anaemia. AB - A number of systemic autoimmune diseases are associated with increased levels of the agalactosyl (G0) IgG isoforms that lack a terminal galactose from the CH2 domain oligosaccharide. The current aim was to determine whether the galactosylation of serum IgG is also reduced in a classic antibody-mediated, organ-specific autoimmune condition, and whether the pathogenic autoantibodies are preferentially G0. In two murine forms of autoimmune haemolytic anaemia (AIHA), sera and autoantibodies eluted from erythrocytes were obtained, and the levels of G0 measured using a lectin-binding assay. Serum IgG galactosylation was unaffected following the induction of AIHA in CBA/Igb mice by immunization with rat erythrocytes, but in all animals with the disease the IgG autoantibodies generated were more G0 than the sera. The anti-rat erythrocyte antibodies were similar to the autoantibodies in being preferentially G0, and when CBA/Igb mice were immunized with canine erythrocytes as a control foreign antigen, there was again a bias towards the production of G0 IgG antibodies. In NZB mice with chronic, spontaneous AIHA, the concentration and galactosylation of both serum IgG and autoantibodies were lower than in the induced model, and the ratio of G0 IgG in the serum and erythrocyte eluates varied markedly between different individuals. Our interpretation of these results is that changes in serum IgG or autoantibody galactosylation are not consistent in different models of AIHA, and that production of low galactosyl antibodies can be a feature of a normal immune response. PMID- 10469047 TI - Molecular analysis of resolving immune responses in uveitis. AB - To identify the cellular immune processes underlying intra-ocular inflammation, aqueous humour was obtained at cataract surgery from 22 patients with clinically inactive uveitis and 24 patients with age-related cataract. mRNA expression for the cytokines IL-1beta, IL-2, IL-4, IL-6, IL-10, IL-12, interferon-gamma (IFN gamma), transforming growth factor-beta (TGF-beta); T cell subsets CD3, CD4, CD8; monocytes and macrophages (CD14); and B cells (CD19) was measured using reverse transcriptase-polymerase chain reaction (RT-PCR) and radiometric analysis. The majority of uveitis patients demonstrated a T cell-mediated inflammatory response, predominately involving a Th1-like cytokine profile with expression of IL-2 and IFN-gamma in 16/22 and 18/22 samples, respectively. These cytokines were present in only a small number of patients with age-related cataract. This Th1 like polarization was supported by an increased expression of CD8 in a number of patients. IL-1beta was expressed in only six uveitic eyes. Only four patients expressed either IL-4 or IL-10 and no patient expressed both. TGF-beta mRNA could be detected in 18/22 uveitis patients and 15/24 controls. IL-12, the paradigmatic Th1-inducing cytokine, was absent in all samples but CD14 was expressed in the majority of patients and controls. CD19 could not be detected in any sample. The cellular infiltrate in the uveitic eyes showed clear evidence of low IL-1 and absent IL-12 expression despite a Th1-like profile and high expression of macrophages. This strongly suggests that the systemic immunosuppressive therapy used prior to surgery in some patients and/or the chronicity of the uveitis had actively suppressed/switched off macrophage function, leading to resolution of T cell activity. PMID- 10469048 TI - Inhibition of dextran sulphate sodium (DSS)-induced colitis in mice by intracolonically administered antibodies against adhesion molecules (endothelial leucocyte adhesion molecule-1 (ELAM-1) or intercellular adhesion molecule-1 (ICAM 1)). AB - We examined the effect of intracolonic administration of anti-adhesion molecule antibodies on DSS-induced colitis in mice. Immunohistochemical staining in mice with colitis showed increased expression of ELAM-1 and ICAM-1 on endothelial cells of vessels in the lamina propria and submucosa at sites of inflamed lesions. Intracolonic administration of anti-ELAM-1 or anti-ICAM-1 antibody decreased bloody stools, anaemia, and histologically evident damage, as well as myeloperoxidase activity and IL-1beta content. We concluded that adhesion molecule expression is important in the development of DSS-induced colitis in mice and that intracolonic administration of anti-adhesion molecule antibodies, especially anti-ELAM-1 antibody, effectively inhibits the colonic inflammation. Intracolonic administration of anti-adhesion molecule antibodies may show therapeutic promise in ulcerative colitis. PMID- 10469049 TI - Interferon-gamma (IFN-gamma) and prostaglandin E2 (PGE2) regulate differently IL 12 production in human intestinal lamina propria mononuclear cells (LPMC). AB - IL-12 modulates Th1 immune response during chronic colitis. Mechanisms regulating IL-12 synthesis in human intestine are poorly understood. The aim of this study was to investigate the effect of IFN-gamma and PGE2 on lipopolysaccharide (LPS) stimulated LPMC IL-12 production. Normal LPMC cultures were run in the presence or absence of IFN-gamma and/or PGE2 before LPS stimulation. To examine the role of endogenous PGE2 on LPS-stimulated IL-12 release, LPMC cultures were added of indomethacin before LPS stimulation. IL-12, IL-10 and IL-8 were measured by ELISA. No IL-12 was detected in either unstimulated or LPS-stimulated LPMC cultures. In contrast, LPMC released IL-8 (650 +/- 125 pg/ml) and IL-10 (75 +/- 25 pg/ml) in response to LPS. Treatment of LPMC with IFN-gamma facilitated LPS stimulated IL-12, whereas it completely abrogated IL-10 production. IL-12 release by LPMC stimulated with IFN-gamma and LPS was significantly inhibited by exogenous IL-10. The addition of PGE2 to IFN-gamma-treated LPMC cultures inhibited in a dose-dependent manner LPS-induced IL-12 secretion. Furthermore, IL 12 was detectable (85 +/- 25 pg/ml) in the supernatants of LPMC cultures treated with indomethacin and LPS. In contrast to the effect on IL-12, PGE2 significantly augmented LPS-stimulated LPMC IL-10 production. However, the inhibition of IL-12 by PGE2 was only partially reversed by anti-IL-10. In a simplified model of LPS tolerance, we finally showed that monocyte-derived macrophages exhibited reduced IL-12 production after repeat LPS stimulation. In these cell cultures, indomethacin abrogated the induction of LPS desensitization. IFN-gamma and PGE2 modulate differently the LPMC responsiveness to LPS in terms of IL-12 synthesis. PMID- 10469050 TI - The influence of age and gender on serum dehydroepiandrosterone sulphate (DHEA S), IL-6, IL-6 soluble receptor (IL-6 sR) and transforming growth factor beta 1 (TGF-beta1) levels in normal healthy blood donors. AB - Dysregulation of IL-6 synthesis is thought to play a role in the development of a number of age-related conditions, such as rheumatoid arthritis, osteoporosis, atherosclerosis, Alzheimer's disease and B cell malignancies. Recently it has been suggested that the production of IL-6 is influenced by the adrenal hormone dehydroepiandrosterone (DHEA) and its sulphated derivative DHEA-S. In humans we investigated the relationship between DHEA-S, IL-6, IL-6 sR and TGF-beta1 in the serum of normal healthy male and female blood donors. Using immunoassay techniques we found that the serum levels of DHEA-S significantly (P = 0.0001) decreased with age in both males and females. Furthermore, mean DHEA-S levels in all age groups were significantly (P = 0.0001) higher in males. Such correlations were not apparent for IL-6 using a standard assay, but a high sensitivity assay revealed that serum IL-6 was significantly (P = 0.0018) positively correlated with age in males only. In addition, serum levels of DHEA-S were significantly (P = 0.048) negatively correlated with serum IL-6, again in male subjects only. In contrast, serum IL-6 sR and TGF-beta1 levels were not correlated with age in either males or females and were not significantly different between the sexes. However, a significant (P = 0.024) negative correlation between DHEA-S and IL-6 sR was found in males. These studies clearly highlight the complex nature of the relationship between these molecules in the ageing process in normal healthy blood donors and demonstrate the need to use high sensitivity assays when measuring IL-6 in apparently healthy individuals under the age of 70 years. PMID- 10469051 TI - Lymphocyte proliferation modulated by glutamine: involved in the endogenous redox reaction. AB - Decreased glutamine concentrations are found during catabolic stress and are related to susceptibility to infections. However, little is known about the mechanism of glutamine modulation of lymphocyte functions. Glutamine is not only an important energy source in mitochondria, but is also a precursor of glutamate, which is used for cellular glutathione (GSH) biosynthesis in lymphocytes. In this study, we investigated the effects of glutamine on the redox reaction during lymphocyte proliferation. Peripheral blood mononuclear cells, obtained from healthy adult volunteers, were cultured and stimulated by phytohaemagglutinin (PHA) in the presence of different glutamine concentrations. Cells were harvested and prepared for analysis of lymphocyte proliferation, cell cycle propagation, intracellular glutathione levels and reactive oxygen species (ROS) production. We found that glutamine supplementation significantly enhanced PHA-stimulated lymphocyte proliferation and propagation of the cell cycle from the G1 to S and G2/M phases. Glutamine also enhanced production of both intracellular ROS and GSH levels in PHA-stimulated lymphocytes. Flow cytometric analysis by the mercury orange staining method showed that glutamine significantly enhanced intracellular non-protein thiols in PHA-stimulated CD4+, but not CD8+ lymphocyte subsets. Furthermore, intracellular GSH detected by monochlorobimane dye probe showed that glutamine enhanced GSH both in PHA-stimulated CD4+ and CD8+ lymphocyte subsets. Inadequate glutamine supplementation resulted in decreased lymphocyte proliferation in association with decreased levels of intracellular GSH. Addition of exogenous GSH significantly enhanced lymphocyte proliferation, whereas blockade of GSH synthesis enhanced ROS production and suppressed lymphocyte proliferation. These results suggest that the modulation of PHA-stimulated lymphocyte proliferation by glutamine is closely related to the maintenance of appropriate intracellular redox status. PMID- 10469052 TI - Infection of human endothelial cells with Staphylococcus aureus induces the production of monocyte chemotactic protein-1 (MCP-1) and monocyte chemotaxis. AB - Bacterial infection coincides with migration of leucocytes from the circulation into the bacterium-infected tissue. Recently, we have shown that endothelial cells, upon binding and ingestion of Staphylococcus aureus, exhibit proinflammatory properties including procoagulant activity and increased intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression on the cell surface, resulting in hyperadhesiveness, mainly for monocytes. The enhanced extravasation of monocytes to bacterium-infected sites is facilitated by the local production of chemotactic factors. From another study we concluded that the locally produced chemokine MCP-1 is important in the recruitment of monocytes to the peritoneal cavity in a model of bacterial peritonitis. In the present study we investigated whether cultured human endothelial cells after infection with bacteria produce and release MCP-1, which in turn stimulates monocyte chemotaxis. We observed that endothelial cells released significant amounts of MCP-1 within 48 h after ingestion of S. aureus. This was dependent on the number and the virulence of the bacteria used to infect the endothelial cells. The kinetics as well as the amount of MCP-1 released by S. aureus-infected endothelial cells differed markedly from that released by endothelial cells upon stimulation with IL-1beta. Supernatant from S. aureus infected or IL-1beta-stimulated cells promoted monocyte chemotaxis which was almost entirely abrogated in the presence of neutralizing anti-MCP-1 antibody, indicating that most of the chemotactic activity was due to the release of MCP-1 into the supernatant. Our findings support the notion that endothelial cells can actively initiate and sustain an inflammatory response after an encounter with pathogenic microorganisms, without the intervention of macrophage-derived proinflammatory cytokines. PMID- 10469053 TI - Use of reconstituted influenza virus virosomes as an immunopotentiating delivery system for a peptide-based vaccine. AB - Immunopotentiating reconstituted influenza virosomes (IRIV) were used as a delivery system for the synthetic peptide-based malaria vaccine SPf66. The reduced SPf66 peptide molecules containing terminal cysteine residues were covalently attached to phosphatidylethanolamine with the heterobifunctional crosslinker gamma-maleimidobutyric acid N-hydroxysuccinimide ester. The SPf66 phosphatidylethanolamine was incorporated into IRIV and BALB/c mice were immunized twice by intramuscular injection with peptide-loaded virosomes. Titres of elicited anti-SPf66 IgG were determined by ELISA. These titres were significantly higher and the required doses of antigen were lower, when mice had been preimmunized with a commercial whole virus influenza vaccine. After preimmunization with the influenza vaccine, SPf66-IRIV elicited far more consistently anti-SPf66 antibody responses than SPf(66)n adsorbed to alum. MoAb produced by four B cell hybridoma clones derived from a SPf66-IRIV-immunized mouse cross-reacted with Plasmodium falciparum blood stage parasites in immunofluorescence assays. All four MoAbs were specific for the merozoite surface protein-1 (MSP-1)-derived 83.1 portion of SPf66. Sequencing of their functionally rearranged kappa light chain variable region genes demonstrated that the four hybridomas were generated from clonally related splenic B cells. Biomolecular interaction analyses (BIA) together with these sequencing data provided evidence for the selection of somatically mutated affinity-matured B cells upon repeated immunization with SPf66-IRIV. The results indicate that IRIV are a suitable delivery system for synthetic peptide vaccines and thus have a great potential for the design of molecularly defined combined vaccines targeted against multiple antigens and development stages of one parasite, as well as against multiple pathogens. PMID- 10469054 TI - An African green monkey lacking peripheral CD4 lymphocytes that retains helper T cell activity and coexists with SIVagm. AB - Natural infection with simian immunodeficiency virus (SIV) is known to occur in the African green monkey (AGM). The actual onset of the disease has not been recognized in SIVagm infected AGM, and the precise reason for such apathogenicity in the AGM remains unclear. We reported previously that AGM peripheral CD4 lymphocytes underwent a peculiar differentiation from CD4+ to CD4- cells after in vitro activation, and we inferred that the AGM does not fall into a fatal immunodeficient state because of the generation of CD4- helper T cells in vivo. To evaluate this possibility, we examined the relationship between CD4 expression and helper T cell activity in the naturally infected AGM. We identified a healthy monkey almost lacking CD4 T cells in the periphery. This AGM showed no signs and symptoms of immunodeficiency and retained a helper T cell activity in antibody production comparable to those of CD4+ AGMs. In addition, SIVagm could be isolated from CD8+ lymphocytes in the CD4- AGM. These observations suggest that a unique host-virus adaptation has developed in the AGM, and may be helpful in explaining the fundamental reason for the apathogenicity occurring in this monkey. PMID- 10469055 TI - Viral load and CD69 molecule expression on freshly isolated and cultured mitogen stimulated lymphocytes of children with perinatal HIV-1 infection. AB - HIV-1 in adults changes the proportion of mitogen-stimulated lymphocytes expressing the CD69 activation molecule, but little is known about this molecule expression on lymphocytes of HIV-1-infected (HIV-1+) children. Freshly isolated CD3+, CD4+, CD8+ and CD19+ and phytohaemagglutinin (PHA)-stimulated CD3+, CD4+ and CD8+ lymphocytes co-expressing CD69 were investigated cross-sectionally (adopting a MoAb double-staining technique) in 24 HIV-1+ children with severe disease and given anti-retroviral therapy and in 24 age-matched healthy children. CD69 results in HIV-1+ children were correlated with plasma HIV-1 RNA load prospectively determined. HIV-1+ compared with healthy children had higher frequencies of freshly isolated CD3+CD69+ (2.4 +/- 2.2% versus 0.9 +/- 0.5%; P = 0.002) and CD8+CD69+ (1.5 +/- 1.1% versus 0. 5 +/- 0.2%; P < 0.0001) lymphocytes. The frequencies of CD4+CD69+ and CD19+CD69+ lymphocytes were similar. High viral load correlated with an elevated proportion of freshly isolated CD3+CD69+ and CD8+CD69+ lymphocytes. HIV-1+ children showed reduced frequencies of PHA stimulated CD3+CD69+ (60.7 +/- 7.6% versus 86.1 +/- 7.6%; P < 0. 001), CD4+CD69+ (73.6 +/- 18.2% versus 92.6 +/- 5.1%; P < 0.001), and CD8+CD69+ (51.0 +/- 19.1% versus 65.3 +/- 15.4%; P = 0.007) lymphocytes. Virologic worsening within 6 months correlated with a low proportion of PHA-stimulated CD3+CD69+ and CD8+CD69+ lymphocytes. CD69 molecule expression reflected the coexistence of immune activation and immune deficiency in HIV-1 infection. Changes partly differed from those observed in HIV-1+ adults. CD8+CD69+ (but not CD4+CD69+) lymphocyte proportion correlated with virologic course, and an impaired ability of CD8+ lymphocytes to express CD69 upon PHA stimulation preceded a virologic worsening. PMID- 10469056 TI - Onchocerciasis modulates the immune response to mycobacterial antigens. AB - Chronic helminth infection induces a type-2 cellular immune response. In contrast to this, mycobacterial infections commonly induce a type-1 immune response which is considered protective. Type-2 responses and diminished type-1 responses to mycobacteria have been previously correlated with active infection states such as pulmonary tuberculosis and lepromatous leprosy. The present study examines the immune responses of children exposed to both the helminth parasite Onchocerca volvulus and the mycobacterial infections, Mycobacterium tuberculosis and M. leprae. Proliferation of peripheral blood mononuclear cells (PBMC) and production of IL-4 in response to both helminth and mycobacterial antigen (PPD) decreased dramatically with increasing microfilarial (MF) density. Although interferon gamma (IFN-gamma) production strongly correlated with cellular proliferation, it was surprisingly not related to MF density for either antigen. IL-4 production in response to helminth antigen and PPD increased with ascending children's age. IFN gamma and cellular proliferation to PPD were not related to age, but in response to helminth antigen were significantly higher in children of age 9-12 years than children of either the younger age group (5-8 years) or the older group (13-16 years). Thus, there was a MF density-related down-regulation of cellular responsiveness and age-related skewing toward type 2 which was paralleled in response to both the helminth antigen and PPD. This parasite-induced immunomodulation of the response to mycobacteria correlates with a previous report of doubled incidence of lepromatous leprosy in onchocerciasis hyperendemic regions. Moreover, this demonstration that helminth infection in humans can modulate the immune response to a concurrent infection or immunological challenge is of critical importance to future vaccination strategies. PMID- 10469057 TI - A switch towards Th2 during serological rebound in children with congenital toxoplasmosis. AB - Serological rebounds occur frequently in patients with congenital toxoplasmosis, but remain poorly understood. A link between Th1 and Th2 cytokines and the pathophysiology of infectious diseases has been reported. Production of interferon-gamma (IFN-gamma) and IL-4 in supernatants of whole blood after in vitro specific Toxoplasma gondii stimulation and serum-specific IgE levels were studied in 31 congenitally infected children. IFN-gamma was produced at higher levels by lymphocytes from children with stable congenital toxoplasmosis (n = 18) than from children showing serological rebound (n = 13) (P < 0.04). Conversely, supernatants from children with serological rebound showed higher levels of IL-4 than those from children with stable congenital toxoplasmosis (P < 0.03). The polarized Th2 response was confirmed by a greater (IL-4:IFN-gamma) x 100 ratio (P < 0.0001) and production of T. gondii-specific IgE in six out of 13 children showing serological rebound. These results suggest a role of Th2 cytokines in destabilization of congenital toxoplasmosis and perhaps in local reactivation of the parasite. PMID- 10469058 TI - Natural killer cells in cross-regulation of IL-12 by IL-10 in Leishmania antigen stimulated blood donor cells. AB - We have previously shown that natural killer (NK) cells play a role in protection against leishmaniasis. Furthermore, we have shown that NK cells in mononuclear cells derived from unexposed donors are induced to proliferate in vitro in response to leishmanial antigens. Since interleukin (IL)-12, a strong inducer of NK cells, acts on the early events in NK cells and T-cells, and is considered as an adjuvant for use in a potential antileishmaniasis antigen, we wished to investigate how this cytokine influences the in vitro Leishmania induced proliferative and cytokine response in healthy donors. We demonstrate that in an innate response to Leishmania antigen involving NK cells, a critical level of IL 12 is required to induce interferon (IFN)-gamma secretion below which, IL-10 is released in amounts which apparently inhibit IFN-gamma secretion and cellular proliferation. However, at higher IL-12 levels, there is simultaneous secretion of IFN-gamma and IL-10 as well as proliferation of cells. In a similar vein, exogenous IL-10 in turn inhibited IFN-gamma secretion as well as proliferation when used at low/medium concentrations, but at high concentrations this effect was abolished and replaced by the simultaneous detection of IFN-gamma, IL-10 and proliferation. The contribution of NK cells in cross regulation of these two very important immuneregulatory cytokines and the effect of exogenous IL-12 in a Leishmania driven response are discussed. PMID- 10469059 TI - Anti-oxidative capacity in patients with ataxia telangiectasia. AB - Highly reactive oxygen species (ROS) are involved in T-cell activation and in the defense against environmental pathogens. An imbalance of ROS generation and detoxifying scavenger enzymes could contribute to the increased susceptibility to cancer and infections in ataxia telangiectasia. We studied oxidative status, i.e. plasma total antioxidant capacity (TEAC), retinol, alpha-tocopherol, ubiquinol, and the number of activated T cells in 10 patients with ataxia telangiectasia (AT) compared to age-matched healthy controls. As expected, patients showed significantly increased levels of activated human leukocyte antigen-DR and CD45RO expressing T cells. TEAC levels as well as the exogenous antioxidants retinol and alpha-tocopherol were significantly reduced in patients. In addition, patients showed slightly reduced plasma levels of the endogenous ROS scavenger enzyme ubiquinol (Q10). Although no correlation between number of activated T-cells and antioxidant capacity could be demonstrated, an increase in ROS and a diminished reactive oxygen scavenger capacity may be involved in the disease process of patients with AT. PMID- 10469060 TI - Colony-stimulating factor-1 (CSF-1) expression in the uteroplacental unit of mice with spontaneous and induced pregnancy loss. AB - CSF-1 plays an important role in female reproduction and normal embryo development. To understand further CSF-1 function in normal and, especially, in compromised pregnancy, we studied the pattern of its mRNA expression as well as expression of its receptor (c-fms) in the uteroplacental units of mice with induced (cyclophosphamide (CY)-treated) and spontaneous (CBA/J x DBA/2J mating combination) pregnancy loss. RNase protection analysis demonstrated the presence of two forms of CSF-1 mRNA in the uteroplacental unit corresponding to 1400- and 263-bp protective fragments. Densitometric analysis demonstrated that the level of 1400-bp mRNA form was decreased by 40% in the uteroplacental units of mice with CY-induced pregnancy loss compared with the control mice. About 20% decrease in 263-bp protective fragment was registered in resorbing versus non-resorbed placenta of CBA/J females mated to DBA/2J males. As judged by in situ hybridization assay, CSF-1 mRNA transcripts were localized in the uterine epithelium and stroma, while c-fms mRNA was found mainly in the trophoblast. The number of metrial gland cells as well as the number of uterine leucocytes expressing CSF-1 and c-fms mRNAs was substantially lower in the uteroplacental unit of mice with pregnancy loss than in control animals. Maternal immunostimulation, while significantly decreasing the resorption rate in mice with CY-induced pregnancy loss, also strengthened CSF-1 mRNA expression at the fetomaternal interface and resulted in reconstitution in the number of CSF-1+ uterine leucocytes and metrial gland cells. These data suggest a role for uterine CSF-1 in the physiology of normal and compromised pregnancy and demonstrate a possible involvement of CSF-1-associated signalling in mechanisms of placenta and endometrium repair following immunopotentiation. PMID- 10469061 TI - Quantitative analysis of peripheral blood Th0, Th1, Th2 and the Th1:Th2 cell ratio during normal human pregnancy and preeclampsia. AB - We calculated the percentage of Th1, Th2, Th0 cells and the Th1:Th2 cell ratio of peripheral blood from normal pregnant subjects and preeclampsia patients using flow cytometry which can analyse both the surface marker, CD4, and intracellular cytokines, interleukin (IL)-4 and interferon (IFN)-gamma. In normal pregnancy, the percentage of Th1 cells was significantly lower in the third trimester, and the ratios of Th1:Th2 were significantly lower in the second and third trimester than in nonpregnant subjects. In contrast, the percentage of Th1 cells and the ratios of Th1:Th2 in preeclampsia were significantly higher than in normal third trimester pregnant subjects. The percentage of Th2 cells in preeclampsia was significantly lower than in third trimester of normal pregnancy. Additionally, peripheral blood mononuclear cells from these subjects and patients were cultured with phytohemagglutinin stimulation, and IL-4 and IFN-gamma concentrations were determined in the supernatant by enzymed linked immunosorbent assays. The percentage of Th1 and Th2, and the ratios of Th1:Th2 were correlated with cytokine (IFN-gamma and IL-4) secretion level. These results demonstrated that Th2 cells were predominant in the second and third trimesters of normal pregnancy, but Th1 cells predominated in preeclamptic patients. PMID- 10469062 TI - Kinetics of serum soluble tumour necrosis factor receptor (TNF-R) type-I and type II after a single interferon-alpha (IFN-alpha) injection in chronic hepatitis C. AB - Circulating soluble TNF receptors, which act as TNF inhibitors, increase following the administration of IFN-alpha. Whether this is due to a direct IFN action or to indirect mechanisms involving the release of other cytokines is unclear. The kinetics of serum IFN, TNF, IL-6, IL-10, soluble TNF receptor type-I (sTNF-RI) and sTNF-RII were evaluated by enzyme immunoassays in 11 patients with chronic hepatitis C, following the first dose of recombinant human IFN-alpha2b (3 MU given subcutaneously). sTNF-RI concentrations paralleled IFN concentrations, rising from a mean +/- s.e.m. value of 3.5 +/- 0.3 ng/ml at baseline to a peak value of 5.5 +/- 0.5 ng/ml after 9 h, followed by a return to 4.1 +/- 0.4 ng/ml after 24 h (P = 0.0001). sTNF-RII concentrations, which were 7.6 +/- 0.5 ng/ml at baseline, fell initially to 6.9 +/- 0.5 ng/ml, to reach a peak at 24 h of 9.0 +/- 0.7 ng/ml (P < 0.0001). In contrast, the concentrations of TNF, IL-6 and IL-10 fluctuated with no significant changes at any time point. The area under the curve (AUC) of incremental IFN values had a strong positive correlation with the AUC of incremental sTNF-RI values (r = 0.75, P < 0.01). In patients with hepatitis C, IFN concentrations reached after a single dose of IFN were paralleled by correlationally increased concentrations of sTNF-RI, which are a much better marker of administered IFN than sTNF-RII, IL-6 or IL-10. PMID- 10469063 TI - Anti-neutrophil cytoplasmic antibodies (ANCA) against bactericidal/permeability increasing protein (BPI) and cystic fibrosis lung disease. AB - Persistent infection with Pseudomonas aeruginosa and inflammatory mechanisms play an important role in cystic fibrosis (CF) lung disease. ANCA against BPI, a potent host defence protein with anti-bacterial and anti-endotoxin properties, have been described in CF. We have assessed the relationship of anti-BPI antibodies to pulmonary disease severity in 148 CF subjects. IgA and IgG anti-BPI antibodies were found in 55.4% and 70.3% of CF patients, respectively, and higher levels were strongly associated with colonization with P. aeruginosa (P = 0.001 and 0.039 for IgA and IgG antibodies, respectively). IgA and IgG anti-BPI antibodies were independently associated with more severe lung disease as assessed by chest radiograph score (P = 0.023) and a significantly lower forced expiratory volume in 1 s (FEV1)% (P = 0.01). The pathophysiological relevance of the autoantibodies was investigated further by determining their epitope specificity and their effect on bacterial phagocytosis in vitro. Both isotypes of anti-BPI antibodies were specific for the C-terminus of BPI shown recently to be important for BPI-mediated opsonization, and in vitro affinity-purified anti-BPI antibodies significantly reduced BPI-induced phagocytosis of Escherichia coli compared with controls. These data indicate that anti-BPI autoantibodies are associated with colonization with P. aeruginosa and worse lung disease in CF. The inhibition of bacterial phagocytosis suggests that these autoantibodies may contribute to the persistence of P. aeruginosa in the CF lung and so play a role in perpetuating CF lung damage. PMID- 10469064 TI - Anti-lactoferrin antibodies and other types of anti-neutrophil cytoplasmic antibodies (ANCA) in reactive arthritis and ankylosing spondylitis. AB - Fifty-five serum samples from patients with reactive arthritis (ReA), 40 from patients with ankylosing spondylitis (AS) and three from patients with chronic sacroiliac joint arthritis were analysed for the presence of ANCA of IgG class by means of enzyme immunosorbent assay using lactoferrin (Lf), myeloperoxidase (MPO) and antigen extracted from azurophil granules ('alpha-antigen') containing proteinase 3 (PR3) as substrate. IgG-ANCA were found in 31 (56%) patients with ReA. Twenty-three (42%) had anti-Lf antibodies, nine (16%) had anti-MPO and eight (15%) had anti-alpha-antigen antibodies, none of which reacted with PR3. Only six (14%) AS or sacroiliac joint arthritis patients had ANCA (P < 0.001). Three (7%) had anti-Lf, two (5%) anti-MPO and two (5%) anti-alpha-antigen antibodies. Yersinia and Salmonella bacteria were separated by SDS-PAGE and blots were incubated with serum from rabbits immunized with human Lf. The hyperimmune serum recognized a band of 78 kD from both bacteria which was not seen when preimmune serum was used. The reaction to the 78-kD antigen could be completely inhibited when anti-Lf antibodies were absorbed on Lf coupled to cyanogen bromide-activated Sepharose, possibly indicating cross-reacting epitopes in Lf and enterobacterial antigen. PMID- 10469065 TI - Isotype distribution and clinical relevance of anti-beta2-glycoprotein I (beta2 GPI) antibodies: importance of IgA isotype. AB - The aim of this study was to evaluate the prevalence of IgG, IgA and IgM anti beta2-GPI antibodies in anti-phospholipid syndrome (APS), and to establish the clinical significance of IgA type antibodies compared with the other isotypes. Anti-beta2-GPI antibodies were measured in the sera of 70 patients by solid-phase enzyme immunoassay in gamma-irradiated polystyrene plates coated with human purified beta2-GPI. Thirty-three out of the 70 patients were classified as having APS: three of them had primary, and 30 had secondary APS related to systemic lupus erythematosus (SLE). The remaining 37 patients had SLE without APS. Anti beta2-GPI antibodies of IgG, IgA and IgM isotypes were present in 84.8%, 59.3% and 51.5% of patients with APS. Both the frequency and the level of each isotype were significantly higher in patients with APS. This association was very strong for IgA (P = 0.0004 for the antibody frequency and P < 0.0001 for the antibody level), as well as for IgG type antibodies (P < 0.0001 and P < 0.0001), whereas it was weaker for IgM (P = 0.01 and P = 0.04). A strong relationship was demonstrated between increased IgA anti-beta2-GPI antibody levels and a history of venous thrombosis, thrombocytopenia, heart valve disease, livedo reticularis and epilepsy. IgG anti-beta2-GPI antibodies were associated with the presence of lupus anticoagulant (LA) in addition to the main features of APS. However, antibodies of IgM isotype were related only to thrombocytopenia and heart valve disease. We recommend the evaluation of anti-beta2-GPI antibodies of IgA isotype in addition to IgG in patients with clinical suspicion of APS. PMID- 10469066 TI - Is an epitope on keratin 17 a major target for autoreactive T lymphocytes in psoriasis? AB - Psoriasis is a T cell-mediated inflammatory skin disease that has been associated with infections by group A beta-haemolytic streptococci. In a previous study of patients with active psoriasis we demonstrated an increased frequency of circulating Th1-like cells that responded to 20 amino acid (aa) streptococcal M peptides sharing sequences with human keratin. These cells disappeared after ultraviolet B (UVB)-induced clinical remission. Using T cells from the blood of 17 psoriatic patients and 17 healthy controls we have now compared the numbers of interferon-gamma (IFN-gamma)-producing cells induced by seven 18-20 aa keratin peptides and five corresponding M-peptides. The most frequent and strongest responses were observed to a peptide from keratin 17 that shares ALEEAN sequence with M-protein. The responses to this peptide were stronger than to the corresponding M-peptide containing the ALEEAN sequence. After UVB treatment T cell responses to all the M- and keratin peptides were abolished, while responses to the positive control antigen streptokinase/streptodornase (SK/SD) were not affected. These findings are consistent with the notion that aa sequences which keratin has in common with M-protein may be a major target for autoreactive T cells in psoriasis. PMID- 10469067 TI - Quick recovery in the generation of self-reactive CD4low natural killer (NK) T cells by an alternative intrathymic pathway when restored from acute thymic atrophy. AB - The thymus comprises the mainstream of T cell differentiation which produces conventional T cells and an alternative pathway which produces primordial T cells with intermediate density of T cell receptor (TCR)-CD3 complex on the surface (i.e. intermediate TCR cells or TCRint cells). We induced acute thymic atrophy in mice by an administration of hydrocortisone (10 mg) or irradiation (6.5 Gy). It was demonstrated that CD3intCD4lowNK1.1+ T cells were immediately generated by an alternative intrathymic pathway without passing through the double-positive CD4+8+ stage, when restored from thymic atrophy (days 3-14). These CD3intCD4lowNK1.1+ T cells mediated self-reactivity and appeared even in the periphery. mRNA of an invariant chain of TCR Valpha14Jalpha281 gene product was detected in these CD4low T cells, but not remaining CD4high T cells. The mainstream of T cell differentiation in the thymus was not restored up to day 14 and there was no leakage of self-reactive clones into the population generated through the mainstream. These results reveal that an alternative intrathymic pathway is associated with the generation of self-reactive T cells, in an early restoration phase after thymic atrophy. PMID- 10469068 TI - Spleen autotransplantation provides restoration of functional splenic lymphoid compartments and improves the humoral immune response to pneumococcal polysaccharide vaccine. AB - After splenectomy, patients have an increased risk of overwhelming post splenectomy infection (OPSI) or sepsis involving encapsulated bacteria such as pneumococcus. The value of spleen autotransplantation after splenectomy because of trauma has long been questioned. Much attention has been given to the restoration of mononuclear phagocyte system (MPS) function, which appeared to be similar to that of splenectomized individuals. The presence of specific anti pneumococcal antibodies may enhance phagocytosis of opsonized bacteria by other parts of the MPS, as present in the liver. Therefore, in the present study we have evaluated the restoration of the humoral immune response after spleen autotransplantation, especially to pneumococcal capsular polysaccharides (PPS). Wistar rats were divided into three groups which were operated as follows: splenectomy, splenectomy followed by autotransplantation, and sham operation. After 12 weeks the rats were vaccinated with 23-valent pneumococcal vaccine. Blood samples were taken after 3 days, 3 and 6 weeks for anti-PPS IgM and IgG ELISA against types 3, 4, 6, 9, 14 and 23. In addition, immunohistological studies were performed on the autotransplants. Significant antibody titre rises were found in a main proportion of the autotransplanted rats, comparable to sham operated rats. Splenectomized rats showed as well a significantly lower increase in immunoglobulin levels, as significant differences in the proportion of rats showing a minimum two-fold increase of antibody level, considered to represent an adequate response. The titres were highest 3 days after vaccination. Immunohistochemical studies demonstrated structurally functional autotransplants, including an intact marginal zone. Considering this significant anti- pneumococcal antibody response, spleen autotransplants can be expected to enable an improved humoral response to PPS, and to contribute to protection against OPSI after splenectomy. PMID- 10469069 TI - Does the location of the surgery or the specialty of the physician affect malpractice claims in liposuction? AB - BACKGROUND: There is increasing national dialogue on who should perform liposuction and where it should be performed. OBJECTIVE: To determine the effect of the location of liposuction surgery and the specialty of the physician on the incidence of malpractice claims. METHODS: Physicians Insurance Association of America malpractice data from 1995-1997 was analyzed. RESULTS: Hospital-based liposuction had more than 3 times the rate of malpractice settlements than office based liposuction. Dermatologists accounted for less than 1% of malpractice claim settlements in liposuction. CONCLUSION: Dermatologic liposuction education has emphasized small volume cases performed under local anesthesia using the tumescent technique. The safety of this approach appears to be validated in terms of decreased malpractice settlements. PMID- 10469070 TI - Multi-electrode radio frequency resurfacing of ex vivo human skin. AB - BACKGROUND: A novel radio frequency multi-electrode surgical (MES) device has produced minimal residual thermal collagen damage (RTCD) and rapid healing in treating articular cartilage. OBJECTIVE: To assess the immediate tissue effects of the MES device on human skin. METHODS: Specimens of ex vivo human skin were subjected to 1 to 3 passes of the device at 3 voltage settings, then processed for routine and polarized light microscopy. The degree of tissue ablation, amount of RTCD, and level of follicular injury were assessed. RESULTS: Epidermal ablation did not occur; epidermal removal correlated with manual debridement after the first passes. Dermal ablation was not readily discernable even after 2 or 3 passes. The thickness of RTCD tended to increase with increased pass number. The mean thickness of this zone of damaged collagen measured 24-53 micro after 1, 63-83 micro after 2, and 80-97 micro after 3 passes, respectively. For a given number of passes, the thickness of the zone of RTCD did not consistently change with increasing voltage applied (86, 108, and 139 V). The small number of samples did not enable meaningful statistical analysis of these observations. In samples with pilosebaceous units, the electrosurgical keratinocyte injury extended down the follicular epithelium, but never beyond the level of the infundibulum. CONCLUSIONS: These findings suggest the MES device produces immediate tissue effects favorable for effective cutaneous resurfacing and rapid postoperative wound healing. PMID- 10469071 TI - Resurfacing the acne-scarred face. AB - BACKGROUND: : Dermabrasion has been a useful method for the improvement of acne scars since 1953. However, the improvement is often limited. Adjuvant procedures are often necessary to improve results. OBJECTIVE: : To improve the results of resurfacing the acne scarred face. By combining the techniques of subcutaneous filling, laser shrinkage of collagen, dermabrasion, and excision, we hope to achieve better results. METHODS: A Jessner/TCA peel is performed on the neck and decollete area after the skin has been preconditioned with vitamin A conditioning lotions. The acne scars are subcised with a semi-blunt needle, and the developed pockets are filled with adipose tissue. Following this, the surface skin is vaporized with three passes of the CO2 laser, and the deeper acne scars in the mid-face region are sanded with a diamond fraise. Residual scars are excised and sutured. A semi-occlusive dressing is used for 5 days, then replaced with an ointment-based moisturizer. After 10 days, a moisturizer-sunscreen is used, followed with a bleaching cream at 15 days. Make-up may be applied after 14 days. CONCLUSION: : By combining these multiple modalities it is possible to produce a dramatic improvement of the acne-scarred complexion. PMID- 10469072 TI - Laser resurfacing: usual and unusual complications. AB - The use of the carbon dioxide laser for skin resurfacing was initially described in 1989. 1 Since that time, several reports have shown it to be highly effective in the treatment of photodamaged skin and acne scarring. 2,3,4,5 Advances in laser technology have simplified the procedure and minimized adverse sequelae. Laser skin resurfacing has become a very popular technique, and recently several patient series have been published on the use of different resurfacing lasers to treat photodamaged skin. 3,4 However, very little has been written about its complications. Adequate patient selection, sound medical judgement, proper training with experience and knowledge of skin physiology and wound care are important factors for successful outcomes. Interested physicians across a broad range of subspecialties have expressed concern about the rate of adverse outcomes and management of complications. We report seven representative cases of complications referred to our dermatology clinics from outside physicians, in the hope of educating clinicians regarding the usual and unusual side effects of this procedure. PMID- 10469073 TI - Comparison of the 585 nm pulse dye laser and the short pulsed CO2 laser in the treatment of striae distensae in skin types IV and VI. AB - BACKGROUND: Striae distensae, or stretch marks, are a very common cosmetic problem. The successful management of stretch marks has long been a source of frustration and curiosity for both the clinician and the researcher. Recent studies suggest lasers may have a role in their management. As yet, no study has reported on the effects of either of these lasers in the treatment of stretch marks on persons with skin types 4 to 6. OBJECTIVE: To assess the efficacy of short pulsed CO2 laser and pulsed dye laser for the treatment of stretch marks in skin types 4, 5, and 6. METHODS: Four patients were initially enrolled in the study. All were female with abdominal stretch marks, present for a range of 8 to 19 years. The patients had skin types ranging from 4 to 6. The test area consisted of a stretch mark long enough to be divided into three contiguous 2 cm sections, labeled A, B, and C. Section A served as the short pulsed CO2 test site, section B served as a control, while section C served as the 585 nm pulsed dye site. Patients were seen for evaluation after 1 week, then every 4 weeks for a total of 20 weeks. Patients were evaluated subjectively by the investigators, and the patients' own self-evaluation was reported as well. RESULTS: Following the 585 nm pulse dye laser, at 20 week follow-up patients with type 4 skin showed no improvement, while type 6 skin showed hyperpigmentation. The short pulsed CO2 test site showed persistent erythema in type 4 skin and marked hyperpigmentation in type 6 skin. CONCLUSIONS: For patients with types 4, 5, and 6 skin, laser treatment of striae should be avoided or used with great caution. PMID- 10469074 TI - Tumescent anesthesia in ambulatory phlebectomy: addition of epinephrine. AB - BACKGROUND: The advantages of using tumescent anesthesia for ambulatory phlebectomy have recently been described. Previously, tumescent solutions have avoided epinephrine for concerns of toxicity given the large volume of anesthetic sometimes used. OBJECTIVE: To evaluate the efficacy and safety of using epinephrine in the tumescent anesthesia solution during ambulatory phlebectomy. METHODS: Over the course of 1 year, epinephrine in the concentration of 1:100,000 was added to the tumescent solution of patients undergoing ambulatory phlebectomy. A retrospective review of 94 sequential patients was performed to determine the rate of complications associated with the procedure. RESULTS: The complication rate was considerably improved using tumescent lidocaine with epinephrine compared to a previously performed study of tumescent lidocaine without epinephrine. The rate of hematoma was decreased to nil while the rate of hyperpigmentation decreased from 3.6% to 0%. Overall, the rate of complications was improved when epinephrine was added to the tumescent lidocaine solution. Blood pressure measured every 5 minutes and heart rate measured continuously did not significantly change before, during or after infiltration of the anesthetic solution. CONCLUSIONS: Epinephrine in appropriate concentrations is clearly safe when used in the tumescent anesthetic solution during ambulatory phlebectomy and should be used to reduce the incidence of hematoma and hyperpigmentation. PMID- 10469075 TI - Raising eyebrows with botulinum toxin. AB - BACKGROUND: Brow elevation rejuvenates the facial appearance. OBJECTIVE: To determine if a significant degree of brow elevation could be achieved through selective botulinum toxin treatment of brow depressors. METHODS: Seven women aged 31-42 (mean 37) years old were treated. The distance from lowest eyebrow cilium of the eyebrow to the midpupillary point was measured before and 1 month posttreatment. Botulinum toxin was injected into the glabellar area (7-10U) and the supralateral eyebrow (0-2.5U each side), to a total dose of 10-14 U. RESULTS: Five individuals (71%) showed brow elevation of 1-3 mm with a mean elevation of 1 mm. Two individuals showed no change. Concurrent weakening of the frown response was noted in all patients. CONCLUSION: Botulinum toxin treatment of brow depressors produces a small degree of brow elevation in the majority of patients. PMID- 10469076 TI - Pulsed dye laser treatment of warts: an update. AB - BACKGROUND: Warts are a therapeutic challenge. New studies indicate that pulsed dye laser therapy may be effective, with clearance rates of 72 to 93%. OBJECTIVE: To determine clearance rate in pulsed dye laser treatment of warts and compare our rate to those of other published studies. METHODS: Thirty-three patients with 96 warts received pulsed dye laser treatment for recalcitrant plantar, digital, peri- and subungual, and body warts. RESULTS: Forty-eight percent of patients had complete wart clearance; 45% partially cleared. Sixty-nine percent of those who cleared remained wart-free for an average of 11 months. Mean fluence was 9.4 J/cm2, with an average of 3.4 treatments. Body and palmar warts responded best, digital and peri- and subungual next, and plantar lesions worst. No significant side effects were observed. CONCLUSION: Pulsed dye laser is an effective treatment option for recalcitrant warts with an excellent side effect profile. However, our response rates were not as high as those previously reported, and we feel that further studies would be useful. PMID- 10469077 TI - Sclerotherapy for leg telangiectasia--a blinded comparative trial of polidocanol and hypertonic saline. AB - BACKGROUND: Hypertonic saline (HS) and polidocanol (POL) have been in use around the world for sclerotherapy of telangiectasia for many years. However, despite numerous articles in the literature extolling the virtues of their individual use, few studies scientifically compare their relative efficacies. OBJECTIVE: To compare, in a statistically significant number of female patients, the relative efficacy of hypertonic saline and polidocanol as sclerosants of leg telangiectasia and reticular feeding veins, using each patient as her own control. METHODS: Eighty-one women with roughly matching leg telangiecasia were treated with sclerotherapy. One leg was injected with 20% saline/2% lignocaine, the other with 1% polidocanol, with the patients blinded as to the sclerosant used for each leg. Assessment of percent reduction of vessels, and the complications of matting and hemosiderin staining was conducted at 2 months by 3 methods: the patient's satisfaction, the treating physician's evaluation, and blinded assessment of before and after photographs. RESULTS: There was no statistically significant difference between HS and POL treated legs when assessed clinically or photographically. However, POL caused more staining and matting, and despite patients finding HS more painful at injection, patient satisfaction at follow-up was higher with the HS treated leg. CONCLUSION: 20% HS and 1% POL have equal efficacy in sclerosing leg telangiectasia and reticular feeding veins. POL causes more adverse sequelae, although these may be related to the solution concentration. PMID- 10469078 TI - Histological comparison of postoperative wound care regimens for laser resurfacing in a porcine model. AB - BACKGROUND: The use of short-pulsed CO2 lasers for skin resurfacing is routinely performed, but few studies have examined postsurgical care. OBJECTIVE: To determine which postoperative treatments are most beneficial in promoting optimal healing after laser resurfacing. METHODS: Four pigs received laser resurfacing. The laser sites were randomly left untreated or treated with petroleum-based ointment or dressed with 1 of the following occlusive dressings: hydrocolloid, hydrogel or foam. Biopsies were taken from each treatment group on Days 2, 3, 4, 5, 8, 12, and 19. All samples were stained with hematoxylin and eosin. Each histological slide was evaluated by a blinded investigator. RESULTS: Differences were observed between treatment groups in the amount of cellular infiltrate, presence of necrotic tissue, progression of the epidermal sheet, maturation of the epidermis, presence of rete ridges, and appearance of new collagen. CONCLUSION: Postoperative treatments after laser resurfacing vary in their ability to influence the quality of healing. PMID- 10469079 TI - Diameter reduction of the proximal long saphenous vein after ablation of a distal incompetent tributary. AB - In this study, we measured the diameter of the competent long saphenous vein proximally and distally to the anastomosis of an isolated incompetent tributary in 12 patients. Measurements were made both before and 30-45 days after surgical removal of the tributary. The preoperative diameter of the saphenous vein proximal to the anastomosis was greater than distally. The difference between proximal and distal to the tributary veins sections diminished from 1.59 mm to 0.39 mm (p = 0.0033) after surgical removal of the tributary alone. This phenomenon may be compared with other hemodynamic occurrences between the superficial and deep system. The incompetence of a tributary induces a proximal dilatation of the long saphenous vein. The isolated ablation of an insufficient tributary appears to be a useful measure to prevent degeneration of greater saphenous vein function as this procedure reduces the saphenous diameter. PMID- 10469080 TI - Early clinical results with a multiple synchronized pulse 1064 NM laser for leg telangiectasias and reticular veins. AB - BACKGROUND: The 1064 nm wavelength penetrates tissue and blood vessels with little absorption by melanin. OBJECTIVE: To perform a study examining the effects of 1064 nm laser used in pulses from 4 msec to 16 msec on leg telangiectasias ranging in size from 0.5 to 3 mm. METHOD: In this initial trial, 50 sites on 30 patients were enrolled and treated with a multiple synchronized pulse laser at 1064 nm. The primary parameter utilized was a single 10-16 msec pulse. Improvement was judged by comparison of digital images at 1 month, 2 months, and 3 months posttreatment. Improvement was judged, based on size and number of vessels remaining. Side effects were noted as present or absent at each visit. RESULTS: Immediate contraction or darkening followed by urtication and visible total vessel closure as indicated by absence of blanching and visual elimination of the vessel border occurred in most of the treated sites. Two 3 mm diameter vessels were confirmed to be closed without flow by Duplex ultrasound visualization, using a 10 MHz transducer. Bruising from vessel rupture was seen in approximately 50% of the cases. No epidermal injury was noted in any sites, even in Fitzpatrick skin Types IV. At 3 months follow-up, 75% improvement was noted at treatment sites. CONCLUSIONS: Initial clinical results with a new multiple synchronized pulsed 1064 nm laser indicate that this longer wavelength supplied at pulses of up to 16 msec appears to be a valuable modality for immediate closure and subsequent elimination of leg ectatic veins. Epidermal injury is unlikely, as the near infrared wavelength has minimal interaction with melanin. PMID- 10469081 TI - Clinical response of psoriasis to low-energy irradiance with the Nd:YAG laser at 1320 nm report of an observation in three cases. AB - BACKGROUND: Psoriasis is a perplexing disease for which we have no definitive therapy. many types of lesions and clinical forms exist, from the small, localized indolent papule, to the persistent intractable plaque, to the disabling palmo-plantar involvement to the erythrodermic form. This wide variety of clinical manifestations has made it difficult to apply therapies such as lasers, which can reportedly produce an improvement. Although they have been shown to work, the carbon dioxide laser needs local injection for anesthesia along with healing by second intention, lasting for 3 to 6 weeks, while the flashlamp-pumped dye laser needs to be delivered in individual pulses of 5 to 10 mm in diameter each and repetition rates of only 1 to 2 pulses per second (1-2 Hz). These factors have made both lasers impractical for widespread use given the extensive skin involvement, which is often present. OBJECTIVE: To report on the beneficial effects of this new apparatus in some psoriatic lesions as observed in 3 out of 3 individuals tested. METHODS: Three patients who had lesions ranging from the small indolent papule to the persistent plaque, resistant to conventional treatment, to plantar involvement, underwent focal treatment with low-energy continuous emission of laser light below the threshold of pain for a total of 4 treatments in a span of 2 weeks. RESULTS: A clinical response was observed in all 3 cases. Improvement was noticed at a 4-week follow-up visit and continued for 3 months. Early recurrence in 1 case was noted at the end of this period. Partial response in a case of plantar involvement was seen. CONCLUSIONS: These findings are not only encouraging to undertake a study in a larger number of patients, with several different fluences and treatment frequencies, but are also intriguing regarding the possible mechanism or mechanisms of action. PMID- 10469082 TI - Amelanotic lentigo maligna melanoma: a unique case presentation. AB - Amelanotic melanomas comprise only 2% of melanomas and are commonly a difficult clinical diagnosis, due to the lack of melanin pigment typically found in melanomas. Even rarer is the amelanotic lentigo maligna, which may have an unusual clinical presentation, such as erythema, pruritus, or edema. Biopsy is the key to diagnosis. Multiple therapies for amelanotic lentigo malignas have been tried, but excision, with margin control (Mohs micrographic surgery-frozen or paraffin sections), remains the treatment of choice. PMID- 10469083 TI - Hair grafting in postburn alopecia. AB - BACKGROUND: Hair micrografting has been widely used in male and female androgenetic alopecia but there is little experience with cicatricial or burn alopecia. OBJECTIVE: To evaluate the clinical outcome of a case of postburn alopecia with marked inelastic affected areas but with good elasticity and hair density over occipital donor site, treated with multiple sessions of hair micrografting. METHODS: Single-hair micrografts were employed to reconstruct the hair line but 2- and 3-hair micrografts were implanted over other alopecic areas. Grafts were placed into oblique slits made with N degrees 15 scalpel blade bevelled due to reduced dermis in the recipient area. Donor site was closed with vertical mattress and running suture with 3-0 silk. RESULTS: In 4 sessions 6500 micrografts were implanted into the affected areas, achieving a natural appearing hair line and a good cosmetic result. CONCLUSION: Hair micrografting is a promising restorative technique for inelastic plaques of postburn alopecia. PMID- 10469084 TI - Cutaneous electrosurgery in a patient with a deep brain stimulator. AB - BACKGROUND: Deep brain stimulators are implantable devices with electrical activity used to treat certain movement disorders such as essential tremor and Parkinson's disease. Similar to implantable cardiac devices, use of electrosurgery on patients with these devices may produce adverse effects. CASE REPORT: We describe the effects of electrosurgery on a patient with essential tremor and an implantable deep brain stimulator who required Mohs micrographic surgery to excise a basal cell carcinoma. The patient experienced immediate lancinating "electrical shock" using electrosurgery in the monopolar mode. The patient experienced no discomfort when a bipolar electrosurgical device was used or when his deep brain stimulator was "turned off." Appropriate positioning of the dispersive plate also reduced adverse effects. CONCLUSION: Dermatologic surgeons should be aware of patients with devices implanted in the CNS with electrical activity and proceed with caution when using electrosurgery. Different approaches can be utilized to help reduce adverse effects. PMID- 10469085 TI - Eccrine syringofibroadenoma treated with a dual pulse width flashlamp pumped pulsed dye laser. AB - BACKGROUND: Eccrine syringofibroadenoma is a rare benign eccrine ductal proliferation with a predilection for the extremities, most often found in middle aged and elderly patients. Reported treatments have included excision and conventional destructive modalities; however, recurrences may be common. OBJECTIVE: We describe a patient with a chronic ulcerated verrucoid eccrine syringofibroadenoma that persisted despite cryotherapy or curettage and electrodessication. RESULTS: Treatment with a dual pulse width flashlamp pumped pulsed dye laser (FPDL) produced an excellent clinical response. CONCLUSION: Treatment of eccrine syringofibroadenoma with a FPDL may provide both a vascular specific injury, analogous to the treatment of verruca vulgaris, and nonspecific thermal destruction at high fluences. PMID- 10469086 TI - Response to Dr. Korc's letter on Mohs surgery. PMID- 10469087 TI - Laser hair removal: a review and report on the use of the long-pulsed alexandrite laser for hair reduction of the upper lip, leg, back, and bikini region. AB - BACKGROUND: The mechanism and permanence of laser-assisted hair removal remains a formidable task in the medical community. OBJECTIVE: The purpose of this study was to determine the safety and long-term efficacy of the long-pulsed or normal mode alexandrite infrared laser for hair depilation. METHODS: Beginning in October 1996, a total of 31 anatomic sites on 22 patients ranging in age from 25 to 59 years (mean 42 years) were evaluated to assess hair removal. Treatment sites included 17 upper lips, 9 legs, 2 backs, and 3 bikini regions. Eligible patients were of Fitzpatrick skin types I-III. Patients were treated using the long-pulsed alexandrite infrared laser at 755 nm, single-pulse technique, 10 mm spot size, 10% overlap, pulse durations of 5, 10, and 20 msec, and a fluence of 20 J/cm2. Subjective patient improvement and objective, blinded graded improvement was assessed at 1, 2, 3, and 6 months. RESULTS: Objective blinded grading at 6 months revealed that hair reduction varied both with the pulse duration and anatomic location. Maximum reductions observed were 40%, 56%, 50%, and 15% for the lip, leg, back, and bikini areas, respectively. Upper lip hair reduction increased from 40% to 54% at 6 months when a second treatment was performed 8 weeks after the initial treatment. CONCLUSION: The long-pulsed alexandrite laser is safe and effective in reducing hair growth. Treatment efficacy varies with the anatomic location, pulse duration, and number of treatments. A single-pulse technique utilizing a 10 msec pulse duration at 20 J/cm2 produced the greatest hair reduction. No permanent adverse effects occurred on skin types I-III at the parameters tested. PMID- 10469088 TI - Unwanted body hair and its removal: a review. AB - Despite widespread demand for efficient, reliable methods of eliminating unwanted hair from the face and body, available options were limited until the recent development of laser-assisted hair removal systems. This is a review of the various types of hair removal methods available today with an emphasis on laser assisted hair removal. PMID- 10469089 TI - Treatment of facial rhytids with a nonablative laser: a clinical and histologic study. AB - BACKGROUND: Current facial resurfacing modalities (laser, chemical peels, and dermabrasion) remove the epidermis and thus cause open wounds which carry significant risks and extended recovery periods. The nonablative laser is a novel Nd:YAG system designed to alleviate facial rhytids without injuring the epidermis. This new modality may offer patients rhytid removal without the risk seen in currently used resurfacing techniques. OBJECTIVE: To describe a novel nonablative laser system and assess its safety profile and efficacy. METHODS: Ten patients received laser treatments of their periocular rhytids and postauricular skin. Clinical variables (rhytid severity, hyperpigmentation, hypopigmentation, scarring, and level of discomfort) were assessed at 1 and 3 months posttreatment. Postauricular skin biopsies were taken both pre- and posttreatment for histologic analysis. RESULTS: Patient discomfort was minimal. Three months posttreatment, 4 of 10 patients showed a one-point improvement in periocular rhytid severity when judged on a six-point scale. The results were not statistically significant. Biopsy analysis showed a small posttreatment increase in the amount of dermal collagen in three patients. A small decrease in collagen was noted in one patient. Three patients also showed a small posttreatment increase in the degree homogenization of dermal collagen. No change from baseline was noted in other assessed histologic parameters. Complications observed included hyperpigmentation in three patients and pitted scarring in three patients. CONCLUSIONS: The nonablative laser gave clinically subtle and statistically insignificant improvement in rhytid severity. Unfortunately its use was associated with complications that included hyperpigmentation and scarring. This technology may eventually offer patients a new resurfacing option, but its efficacy and complication rate must be improved first. PMID- 10469090 TI - Comparison of the 595 nm long-pulse (1.5 msec) and ultralong-pulse (4 msec) lasers in the treatment of leg veins. AB - BACKGROUND: Although several lasers and light sources are now available for vascular lesions, treatment of leg veins has not been very satisfactory. Lengthening the pulse width should theoretically result in improved response rates. OBJECTIVE: This study compared the efficacy and safety of 595 nm pulsed lasers at 1.5 msec and 4 msec in treating leg veins. METHODS: For group A, 27 healthy adult volunteers with leg veins measuring less than 1 mm in diameter were treated with a 2 mm x 7 mm elliptical handpiece. Each patient had three areas treated. The first two areas were treated with the 4 msec pulsed dye laser with fluences of 16 and 20 J/cm2, while the last area was treated with a 1.5 msec pulsed dye laser at fluences ranging from 14 to 16 J/cm2. For group B, 13 subjects were treated. Areas 1 and 2 were treated with the 4 msec pulsed dye laser using a 3 mm x 5 mm and 5 mm handpiece, respectively, while the third site was treated with a 1.5 msec laser using a 3 mm x 5 mm handpiece. Fluences ranging from 14 to 16 J/cm2 were used. Clinical evaluations were performed and photographs taken at 4-8 week intervals. RESULTS: Neither laser regularly induced satisfactory diminution or disappearance of these vessels after one treatment. In group A, more than 50% of patients had little to no improvement, while in group B little to no improvement was observed in 33% of patients following a single treatment. Although there was no significant difference in outcomes between the test sites, the 4 msec PDL with a 3 mm x 5 mm spot size appeared to be most effective. Transient hyperpigmentation was common following either treatment, while hypopigmentation was seen in group B subjects. No scarring was noted. CONCLUSION: Both the 4 and 1.5 msec flashlamp-pumped pulsed dye lasers were ineffective in treating leg telangiectasia following a single treatment. PMID- 10469091 TI - The use of chemical peelings in the treatment of different cutaneous hyperpigmentations. AB - BACKGROUND: Several chemical agents including hydroquinone, retinoic acid, and azelaic acid are currently used in the treatment of cutaneous hyperpigmentations. Recently chemical peelings with kojic acid, glycolic acid, and trichloroacetic acid, either alone or in combination, have been introduced for treatment of hyperpigmentations. OBJECTIVE: The purpose of our study was to evaluate the efficacy of trichloroacetic acid as well as glycolic acid associated with kojic acid in the treatment of cutaneous hyperpigmentations. METHODS: Twenty patients with diffuse melasma were treated with a solution composed of 50% glycolic acid and 10% kojic acid whereas 20 patients with localized hyperpigmentations (lentigo) were treated with 15%-25% trichloroacetic acid. RESULTS: Complete regression of diffuse melasma was observed in 6 of 20 patients (30%), a partial regression in 12 of 20 patients (60%), and no regression in 2 of 20 patients (10%) treated with 50% glycolic acid and 10% kojic acid. Complete regression of localized hyperpigmentations was observed in 8 of 20 patients (40%), a partial regression in 10 of 20 patients (50%), and no regression in 2 of 20 patients (10%) treated with 15-25% trichloroacetic acid. CONCLUSIONS: Based on our findings, both peelings can be considered effective in the treatment of cutaneous hyperpigmentations. PMID- 10469092 TI - Modification of subcutaneous adipose tissue by a methylxanthine formulation: a double-blind controlled study. AB - BACKGROUND: Excessive subcutaneous adipose tissue is typically treated by physically removing the fat through liposuction, but cost and accessibility have popularized alternative treatments for reducing adipose tissue thickness. OBJECTIVE: The purpose of this study was to test the absolute and relative effectiveness of a liposome-encapsulated caffeine-based cream in modifying subcutaneous adipose tissue. METHODS: Forty-one patients consented and completed the double-blind, single-center, placebo-controlled study. Caliper measurements, tape measurements, and photographs were taken over a 2-month period. RESULTS: Both concentrations of the cream were found to significantly reduce the thickness of the adipose tissue in all areas of the body. In addition, the more concentrated cream was significantly more effective than the less concentrated cream in the areas of the hips and the triceps. CONCLUSION: The caffeine-based liposome-encapsulated cream significantly reduced the thickness of the subcutaneous fat over a 2-month period. PMID- 10469093 TI - A comparison of four frequency-doubled Nd:YAG (532 nm) laser systems for treatment of facial telangiectases. AB - BACKGROUND: Telangiectases are cosmetically concerning for millions of individuals who develop them on the face as they mature. The causative factors are numerous. Lasers have recently become the focus for treating small facial telangiectases because of their ability to selectively target vessels. OBJECTIVE: The purpose of this study was to compare the efficacy of four different frequency doubled neodymium:yttrium-aluminum-garnet (FD Nd:YAG) laser (532 nm) systems for the treatment of facial arteriolar telangiectases. METHODS: Forty adult subjects with Fitzpatrick skin phenotypes I and II were randomly assigned to four treatment groups. Each group, consisting of 10 subjects, had telangiectases less than 1000 microm in diameter treated with one of four different FD Nd:YAG laser systems with a wavelength of 532 nm. Patients were treated once and clinical improvement and potential adverse effects were assessed at 1 and 8 weeks after treatment. RESULTS: All treated patients showed improvement after one laser session. No patients were seen to have scarring or pigmentary changes. There was no significant difference in the clinical response following treatment with each laser. CONCLUSION: The four different FD Nd:YAG lasers performed equally well in treating facial telangiectases. With a wavelength of 532 nm and pulse durations in the millisecond range, vessels can be selectively targeted with a minimal incidence of post-operative complications. PMID- 10469094 TI - Treatment of upper lip wrinkles: a comparison of the 950 microsec dwell time carbon dioxide laser to manual tumescent dermabrasion. AB - BACKGROUND: High-energy pulsed or computer-scanned continuous-wave carbon dioxide (CO2) laser resurfacing has gained popularity as a wrinkle treatment because of its minimal thermal injury and precise control of tissue vaporization depth. Manual tumescent dermabrasion has also been effective for treating facial wrinkles. This is, to our knowledge, the first study comparing the use of CO2 laser to manual tumescent dermabrasion for the treatment of wrinkles on the upper lip. OBJECTIVE: To compare prospectively the clinical efficacy of the 950 microsec dwell time CO2 laser to that of manual tumescent dermabrasion in the treatment of upper lip wrinkles. METHODS: Twenty female subjects with moderate to severe upper lip wrinkles were randomly treated with the 950 microsec dwell time CO2 laser on one side of the upper lip and manual tumescent dermabrasion on the other. RESULTS: The average upper lip laser-treated wrinkle score (0 = none to 5 = severe) decreased from 4.3 +/- 0.2 before treatment to 1.8 +/- 0.3 at 6 months after treatment. The average upper lip dermabrasion-treated wrinkle score decreased from 4.4 +/- 0.2 to 1.5 +/- 0.3. The degree to which the wrinkle score improved after laser treatment compared with that after dermabrasion was not statistically significant (P =.216). CONCLUSION: Manual tumescent dermabrasion and 950 microsec dwell time CO2 laser resurfacing are equally effective for the treatment of upper lip wrinkles. PMID- 10469095 TI - Hair restoration surgery in patients with hypotrichosis of the pubis: the reason and ideas for design. AB - BACKGROUND: Hypotrichosis of the pubis is not an uncommon condition, especially in oriental women. Besides the aesthetic problem, this condition may cause low self-esteem, social embarrassment, and psychologic problems to patients. There have been many efforts to correct this condition medically and surgically for decades. Among them hair restoration surgery is thought to be the only definitive therapeutic modality at present. OBJECTIVE: Our purpose was to show the importance of preoperative evaluation of what type of pubic hair patterns the patients may seek and thereafter to make a design based on the patients' desire and physiologic feature of pubic hair for the natural-appearing results and satisfaction of the patients. METHODS: Ten female patients were enrolled in this study aged between 23 and 48 years with pubic hair maturity index class I-III. For selection of a patients' favored pubic hair pattern, we provided photograph samples of pubic hair patterns that consisted of four types as previously documented: horizontal, sagittal, acuminate, and disperse. We restored hairless mons using a conventional one- to three-haired mini-micrograft technique. RESULTS: Five patients belonged to the pubic hair maturity index class I, four to class II, and one to class III. Seven of 10 patients wanted a horizontal (inverted triangular) type, which is most commonly seen in young females, 2 patients wanted acuminate, and 1 wanted sagittal. Eight patients underwent a single-session operation, while two others underwent operations twice. The two patients requiring two operation sessions belonged to a group of class I pubic hair and desired acuminate-type hair. Most patients were satisfied with the results of their operations. CONCLUSION: In designing a pubic hair graft, it is important to know the patients' desire and to make a design based on it for the satisfaction of the patients and for natural-appearing results. Before the procedure, dermatologic surgeons should have to consider a grafted hair line, the distribution, density, and directions of the hair shaft, and the angling of the hair to the skin. PMID- 10469096 TI - Sociodemographic profile and satisfaction with treatment of patients undergoing liposuction in tumescent local anesthesia. AB - OBJECTIVE: To assess patient satisfaction with liposuction in tumescent local anesthesia (TLA) and to define a patient profile regarding sociodemographic data. METHODS: Three hundred randomly selected patients were asked by mail to complete a standardized questionnaire on aesthetic dermatology and cosmetic surgery 3-25 months after liposuction. The 64 items addressed treatment satisfaction, treatment effects on body, social, and professional life as well as self confidence, body feeling, and social contacts. RESULTS: One hundred fifty-nine (53.0%) of 300 patients returned complete questionnaires. Ninety-two percent of the patients were female and the mean age was 45.3 +/- 11.8 years. The mean body mass index (BMI) was 24.5 +/- 3.7, and 61.7% of the patients had a normal BMI. Satisfaction with the liposuction was high (about 85% mostly or completely satisfied). The procedure was regarded by most patients as a nonstressful event (>80%). About 40% reported positive effects on social life, about 91% on body feeling, 57% on attractiveness, and 20% on their profession. CONCLUSION: Liposuction in TLA is a satisfying and well-tolerated treatment for most patients. PMID- 10469097 TI - Comparison of a silicone gel-filled cushion and silicon gel sheeting for the treatment of hypertrophic or keloid scars. AB - BACKGROUND: The exact mechanisms of action responsible for the effectiveness of silicone gel dressings are unknown, although it has been proposed that static electricity generated by friction could be the reason for their anti-scarring effects. OBJECTIVE: We compared the efficacy of a cushion of silicone filled with liquid silicone gel reported to induce greater negative static-electric charge with silicone gel sheeting in the treatment of hypertrophic and keloid scars. METHODS: The size, volume, symptoms (tenderness and itching), and signs (color and induration) of hypertrophic (10 patients) or keloid scars (22 patients) were measured at baseline at 16 weeks following use of either the silicone gel cushion or silicone gel sheeting, as determined by random assignment. RESULTS: Both the silicone gel cushion and the silicone gel sheeting treatments were effective in decreasing scar volume, 53.0% and 36.3%, respectively. The percentages of keloids and hypertrophic scars benefiting from the silicone cushion and the silicone sheeting were similar with respect to reduction in tenderness (36.3% vs 33.3%), itching (45.5% vs 33.3%), and redness (0.1% vs 0.1%), and in the degree of softening (45.5 vs 25.0%). CONCLUSIONS: Both the silicone gel cushion and the silicone gel sheeting treatments were effective in the treatment of keloids and hypertrophic scars, although no statistically significant differences were found between the two treatment modalities. PMID- 10469098 TI - Gabapentin for the treatment of dysesthetic pain after reconstructive surgery. AB - BACKGROUND: Debilitating, postoperative dysesthetic pain after reconstructive surgery is a rare but problematic complication. Conventional therapy is either marginally effective or fraught with side effects. Gabapentin is a new, novel antiepileptic drug helpful in the management of other pain states. OBJECTIVE: A case of disturbing, postoperative dysesthetic pain after reconstructive surgery is presented. METHODS: A trial of gabapentin in escalating dose was initiated and the results and side effects were recorded. RESULTS: Treatment with gabapentin was associated with a substantial decrease in pain, which relapsed after tapering the medication. Relief was obtained with dose escalation, and a successful taper was eventually accomplished. Side effects were minimal. CONCLUSION: This is the first reported use of gabapentin in the treatment of postoperative dysesthetic pain. Although this type of neuropathic pain is difficult to manage, gabapentin produced substantial relief with few side effects. Further investigation is warranted. PMID- 10469099 TI - Subcutaneous leiomyosarcoma on the face. AB - BACKGROUND: Cutaneous leiomyosarcoma is a rare soft tissue sarcoma with a predilection for the lower extremities. Leiomyosarcoma of the face is very rare. Subcutaneous leiomyosarcoma has a higher likelihood of recurrence and metastases than that of the superficial dermal type. OBJECTIVE: The dermatologic surgeon and pathologist should be familiar with the characters of subcutaneous leiomyosarcoma. METHODS: We report a case of subcutaneous leiomyosarcoma on the face with the results of histologic examination and immunohistochemical studies. RESULTS: Leiomyosarcoma of the face is exceedingly rare. The deep subcutaneous type is thought to arise from the smooth muscle of the vascular wall. The neoplasm we report here has deep tumor invasion, high malignancy grade (3B), and large tumor size. Wide excision and postoperative radiotherapy were performed. CONCLUSION: Subcutaneous leiomyosarcoma has a higher likelihood of recurrence and metastases than that of the superficial dermal type. The most effective treatment is wide excision with 3-5 cm lateral margins and a depth that includes subcutaneous tissue and fascia. PMID- 10469100 TI - Prefilled syringes: safe and effective. AB - BACKGROUND: Preloaded syringes are time savers, but questions have arisen anecdotally about the risk of infection from this procedure and the possible loss of potency, especially when performed with buffered syringes. OBJECTIVE: To show that preloaded syringes do not develop colonies of bacterial organisms and to confirm that anesthetic potency is maintained for at least 2 weeks. METHODS: Thirty-six syringes were stored for a period of 2 weeks on a shelf in our clinical procedure area with no protection from heat or light. The majority of these were then cultured for bacteria and fungi and one of them was used on one of the authors to determine the potency of the anesthetic. RESULTS: Preloaded syringes do not appear to be prone to the development of bacterial contamination for at least a 2-week period and potency of the anesthetic is maintained. CONCLUSION: Preloaded syringes are time savers and are a safe modality for use in the practicing dermatology office. PMID- 10469101 TI - Combination treatment of melasma with pulsed CO2 laser followed by Q-switched alexandrite laser: a pilot study. AB - BACKGROUND: Melasma is very difficult to treat and often refractory to treatment with topical creams and pigmented-lesion lasers. OBJECTIVE: Pulsed CO2 laser alone is compared with the combination of pulsed CO2 laser followed by Q-switched alexandrite laser in the treatment of dermal-type melasma. This combination is proposed to be effective by first destroying the abnormal melanocytes with the pulsed CO2 laser and then selectively eliminating the dermal melanin with the alexandrite laser. METHODS: Four patients were randomly chosen for each treatment arm. There were multiple follow-up visits for examination by an objective blinded investigator. RESULTS: All patients in the combination laser group showed complete resolution, and two patients in the CO2 laser only group had peripheral hyperpigmentation in the long-term follow-up evaluation. CONCLUSION: These laser therapies are safe, as there was no scarring and no infection. The combination laser therapy was highly effective in removing the hyperpigmentation and all patients in this group showed complete resolution without any peripheral hyperpigmentation. PMID- 10469102 TI - Primary carcinosarcoma of the skin: report of a case and review of the literature. AB - BACKGROUND: Carcinosarcoma is a biphasic tumor composed of intimately admixed epithelial and mesenchymal elements, both of which are malignant. Primary cutaneous involvement is extremely rare. Therefore it has been difficult to characterize the clinical course of carcinosarcoma of the skin. OBJECTIVE: To describe a further case of carcinosarcoma of the skin and to provide a review of the literature. METHODS: The clinical records of the patient and histopathology of the neoplasm are reviewed. In addition, the results of all previously reported cases of carcinosarcoma of the skin are compiled for reexamination. RESULTS: After complete excision of the carcinosarcoma, the patient shows no evidence of disease for 9 months. Long-term follow-up is needed because of the risk of recurrence or metastasis that may occur with this neoplasm. CONCLUSION: Carcinosarcoma of the skin appears to be associated with a good prognosis when compared with similar tumors located elsewhere. Metastasis (with or without documented mortality) has been reported in 3 of 16 patients (16.7%). Death from local disease has been reported in one additional patient (6.3%). However, further cases with long-term follow-up are needed in order to better understand the nature of this cutaneous neoplasm. PMID- 10469103 TI - Temporal artery biopsy in a dermatologic surgery practice. AB - BACKGROUND: A positive temporal artery biopsy (TAB) is essential to the diagnosis of temporal arteritis. Both this relatively common disease and its prolonged treatment with corticosteroids are associated with serious sequelae. Therefore, accurate and timely diagnosis is critical. The dermatologic surgery literature lacks a description of this straightforward surgical technique, as TABs are most often performed by ophthalmologists. OBJECTIVE: As a service to our rheumatology department we began performing TABs on a same-day on-call basis in July 1996. METHODS: We have performed 45 TABs in a 22-month period using a simple, safe, time-efficient technique. We review the surgical anatomy and danger zone of the temporal region and potential complications. We describe the biopsy technique which aims at safely obtaining a greater than 2 cm segment of a peripheral branch of the superficial temporal artery (STA), identified preoperatively by doppler ultrasonography. RESULTS: The procedure was performed on the day requested in all cases. Intraoperative time ranged from 20 to 40 minutes. TAB established the diagnosis of temporal arteritis in 8 of 44 biopsies (18%) and in 7 of 35 patients (20%), including 1 of 9 patients in whom we performed bilateral TAB. One patient was diagnosed with small-vessel polyarteritis nodosa by TAB. The mean formalin fixed length of the arterial specimen was 2.2 cm. The length did not vary between positive and negative specimens. There were no complications and the cosmetic results were excellent. CONCLUSION: TAB is a quick, safe, straightforward, and gratifying office procedure which dermatologic surgeons are very qualified to perform. PMID- 10469104 TI - Giant cell fibroblastoma: a variant of dermatofibrosarcoma protruberans treated with Mohs' micrographic surgery. PMID- 10469105 TI - A case report of laser resurfacing as a skin cancer prophylaxis. AB - BACKGROUND: Many patients suffer from multiple nonmelanoma skin cancers. We recently encountered two such patients who requested some form of prophylaxis against future skin cancers. OBJECTIVE: To highlight the potential utility of laser resurfacing as a skin cancer prophylaxis. METHODS: Two patients with histories of multiple facial skin cancers were treated with CO2 laser resurfacing for the purpose of skin cancer prophylaxis. RESULTS: During a follow-up period of 33 and 52 months, both patients have remained free of skin cancers in the main treatment field while developing new tumors outside of this area. CONCLUSION: Laser resurfacing should be considered as a potential method of achieving skin cancer prophylaxis in selected patients. PMID- 10469106 TI - Topical potent corticosteroids for excessive granulation tissue. PMID- 10469107 TI - Hair repair surgery. Corrective measures for improvement of older large-graft procedures and scalp scars. AB - BACKGROUND: Implanted hair follicles generally take and grow well. This can be desirable or undesirable, depending on the skill and aesthetic eye of the surgeon. Older methods employing larger grafts often produced pluggy, bristle brush, or "Barbie doll" hairlines that were unsightly and conspicuous. OBJECTIVE: Since patients possessing these outcomes often seek corrective measures, a search was undertaken to find effective, corrective repair methods. Unfortunately, relatively little has been published on this subject. METHODS AND RESULTS: Discussion will center on recipient site repair, donor site repair, and scar repair. Recipient sites can be improved with a combination of laser ablation, electrolysis, punch removal, and fill-in micrografting. Donor sites can be improved by reexcision, undermining, tissue expansion, and layered closure. Anterior scalp scars may be improved by punch excision and micrografting. CONCLUSIONS: Satisfactory improvement for untoward hair transplantation outcomes may be obtained with one or more corrective procedures in many patients. PMID- 10469108 TI - Superiorly based nasalis myocutaneous island pedicle flap with bilevel undermining for nasal tip and supratip reconstruction. AB - BACKGROUND: Superiorly based flaps with bilevel undermining for the reconstruction of nasal tip and supratip defects have not been utilized to this point for reconstruction of this cosmetic subunit. OBJECTIVE: The purpose of this presentation is to describe a new flap for the reconstruction of nasal tip and supratip defects and to introduce a new method of undermining-bilevel undermining in the raising of these flaps. METHODS: In this presentation we demonstrate (diagrammatically and with fresh cadaver dissection) the methods of preparing, raising, and preforming this new flap. We further introduce and describe the concept of bilevel undermining. RESULTS: We describe the first author's 4-year experience in the development of this flap. We show clinical representative cases with pre- and postoperative results. CONCLUSIONS: Superiorly based myocutaneous island pedicle flaps with bilevel undermining, a new method of nasal tip and supratip reconstruction, yields excellent functional and cosmetic results. Bilevel undermining is a new and valuable method of achieving greater tissue mobility in muscle-based flaps. PMID- 10469109 TI - The scalp biopsy: making it more efficient. AB - BACKGROUND: The scalp biopsy plays an important role in diagnosis and subsequent management of many scalp disorders. This simple diagnostic procedure can be time consuming and frustrating for most dermatologists. OBJECTIVE: To outline an efficient technique for performing a scalp biopsy. METHODS: The technique of performing a scalp biopsy is reviewed and suggestions are made to make this time consuming procedure more efficient. RESULTS: Sufficient specimen for histopathologic evaluation in a short perid of time is obtained. CONCLUSION: Attention to specific details in performing a scalp biopsy allows the physician to obtain a quick and histopathologically informative scalp biopsy. PMID- 10469110 TI - Estrogen and progesterone receptors: their role in postsclerotherapy angiogenesis telangiectatic matting (TM). AB - BACKGROUND: Postsclerotherapy neoangiogenesis telangiectatic matting (TM) occurs in up to 24% of individuals treated by sclerotherapy. Although the etiology is unknown, obstructive flow factors, angiogenic factors, estrogen, inflammatory, and endogenous factors have all been postulated to play a role in its pathogenesis. OBJECTIVE: The aim of the study was to ascertain the presence or absence of estrogen and progesterone receptors in postsclerotherapy TM lesions and thus substantiate their possible role in the pathogenesis of TM. METHODS: Ten women, median age 37.7 years, were included in the study population who developed TM following a single sclerotherapy treatment session employing Sotradecol 0.25% for class I-II telangiectasia/venulectasia. Four of 10 patients had a history of previous hormonal therapy or pregnancy in the previous 12 months prior to entering into the study. Three millimeter punch biopsies were taken at 12 weeks posttreatment and assayed for estrogen and progesterone receptors by the ERICA and PRICA (estrogen/progesterone immune cytochemical assay) techniques. RESULTS: Zero of 10 patients were positive for estrogen/progesterone receptors as assayed by the ERICA/PRICA technique in biopsied sites of postsclerotherapy TM. CONCLUSION: Although estrogen and progesterone may play an indirect role in the development of postsclerotherapy TM via vasodilatory or secondary angiogenic or cytokine release mechanisms, they do not appear to play a primary role in promoting postsclerotherapy neoangiogenesis as demonstrated in this study. PMID- 10469111 TI - Dermabrasion using CO2 dry ice. AB - BACKGROUND: The cosmetic dermatologic surgeon can improve facial scars by using a variety of techniques. Chemical peels, lasers, and dermabrasion are among the most common modalities used. In recent years, laser resurfacing has enjoyed great popularity; however, there is still a role for the time-honored and effective technique of dermabrasion. The recent withdrawal of Freon from the market has made dermabrasion more difficult. OBJECTIVE: To introduce a novel technique of using solid carbon dioxide (CO2) to freeze the skin before dermabrasion. METHOD: Twenty-five consecutive patients with facial acne scars underwent a combination of procedures including chemical peeling and CO2 laser resurfacing, followed by freeze-dermabrasion. Compressed CO2 was then used to make a solid ball of dry ice. The ice was used to freeze the skin prior to dermabrasion. After the application of the dry ice ball to the skin for about 4 seconds, the scares were dermabraded with the large mushroom wheel. As the tissue defrosted, the operator started at the outer edge of the freeze and planed into the center. This procedure was repeated until the desired improvement was achieved. RESULTS: With dry ice, a good skin turgor was achieved. This provided a good foundation for the sanding of the acne scars. The patients were pleased with the results and complications were minimal. CONCLUSION: "Home-made" dry ice works as well as Freon in providing skin turgor for dermabrasion. CO2 dry ice has the added benefits of being inexpensive and environmentally friendly. PMID- 10469112 TI - The complications of blepharoplasty: their identification and management. AB - BACKGROUND: Because of the increasing number of blepharoplasties being done by cosmetic dermatologists, it is necessary to have an increased awareness of sequella and complications. OBJECTIVE: To review the frequent complications following blepharoplasty and to develop methods for their prevention and treatment. METHOD: A thorough history and physical combined with accurate intraoperative markings and postoperative instructions can make the procedure more predictable. CONCLUSION: Armed with an awareness of possible blepharoplasty complications, an adequate history and physical can point to areas of concern. If these guidelines are observed, a blepharoplasty can be completed that keeps both the physician and patient satisfied. PMID- 10469113 TI - The combined use of propofol and fentanyl for outpatient intravenous conscious sedation. AB - BACKGROUND: Intravenous conscious sedation is currently being widely utilized for outpatient surgery including dermatologic surgery. Even though this type of anesthesia is typically administered by a trained, licensed anesthetist, it is important for dermatologists who either intend to or are currently utilizing this type of anesthesia to be familiar with some of the methods and agents that are commonly employed. OBJECTIVE: Propofol and fentanyl are two anesthetic agents that are in prevalent use for skin and soft tissue surgery of brief or limited duration. With the goal of familiarizing dermatologic surgeons with this form of anesthesia, a study was conducted to evaluate the safety and effectiveness of the combination of propofol and fentanyl when used for conscious sedation in an outpatient dermatology center. METHODS: Twenty patients, ages 25-65 years, who required conscious sedation were enrolled. Each patient received a standard dosage of fentanyl and propofol, as determined on a kilogram basis. Sedation time, total procedure time, recovery time, and total propofol dose, along with side effects, were determined. RESULTS: The mean onset to sedation was 52.5 seconds, the mean procedure time was 40 minutes 37 seconds, and the mean interval to recovery was 3 minutes 43 seconds, with a mean total dose of propofol of 5.83 mg/kg. Minimal side effects occurred. CONCLUSION: Propofol when used in conjunction with fentanyl appears to be a safe, quick, and effective method of providing conscious sedation. PMID- 10469114 TI - Recurrent Mondor's disease resolved after exeresis of abdominal lipoma. AB - BACKGROUND: Mondor's disease is a rare disorder characterized by superficial thrombophlebitis, classically of the thoracic-epigastric and/or lateral thoracic veins. The etiopathogenesis of the disease is not entirely clear. OBJECTIVE: We report the case of a patient who presented with recurrent Mondor's disease involving the left abdominal wall along the thoracoepigastric vein as far as the ipsilateral iliac fossa where a lipoma was observed. METHODS: Surgical removal of the lipoma showed a close anatomical relationship between the affected veins and the neoformation. RESULTS: Follow-up after 1 year did not reveal any signs of relapse of the thrombophlebitis. CONCLUSION: Since primary or secondary factors leading to hypercoagulability states were not detected in our patient, we speculate that the interference of venous flow by the abdominal lipoma was probably implicated in the etiopathogenesis of the disease. PMID- 10469115 TI - Bilateral primary mucinous carcinoma of the eyelid treated with Mohs surgery. AB - BACKGROUND: Primary mucinous carcinoma is an uncommon malignant cutaneous tumor which arises most commonly on the eyelid. While rarely causing death, recurrence following primary excision is common and widespread metastasis may occur. OBJECTIVE: We report the first case of bilateral primary mucinous carcinoma of the eyelid. METHODS: A lesion of the left lower eyelid had been resected three times previously with positive conventional margins. Both this lesion and a second primary lesion of the contralateral lower lid were removed with Mohs microscopically controlled excision without recurrence for more than 2 years. CONCLUSION: Multiple lesions of mucinous carcinoma of the eyelid do not necessarily connote metastasis. Mohs microscopically controlled excision may be a suitable form of therapy for primary mucinous carcinoma of the eyelid. PMID- 10469116 TI - Facial laser resurfacing with the propofol-ketamine technique: room air, spontaneous ventilation (RASV) anesthesia. AB - BACKGROUND: Multiple anesthetic approaches exist for full-face laser resurfacing. The propofol-ketamine technique is reviewed as a reasonable alternative to providing adequate anesthesia for full-face laser resurfacing in the office environment. OBJECTIVE: To report outcomes using propofol-ketamine opioid avoidance, room air, spontaneous ventilation monitored anesthesia care (MAC). METHOD: A retrospective chart review of 95 consenting adult patients receiving propofol-ketamine anesthesia in a private practice, office-based setting. RESULTS: An average of 6 (200 mg) ampules of propofol, including waste, were used per patient. All patients received adequate anesthesia as evidence by a lack of movement during surgery. There were no hallucinations, no postoperative nausea or vomiting (PONV), no cardiovascular instability or seizures (clinical signs of lidocaine toxicity), and no hospital admissions for either PONV or pain. CONCLUSION: The propofol-ketamine technique appears to be an excellent alternative anesthetic approach to EMLA cream, tranquilizer-opioid regimens, or general inhalational anesthesia for facial laser resurfacing. PMID- 10469117 TI - The phlebectomy probe: a new and useful instrument for ambulatory phlebectomy. AB - BACKGROUND: Tethering of varicose veins by periadventitial connective tissue can limit the length of vein segment that can be avulsed during ambulatory phlebectomy. A thin, malleable blunt probe is described which can dissect connective tissue adhesions surrounding varicose veins. This probe can be used as an endoluminal probe to localize elusive veins. OBJECTIVE: We describe the use of a new type of phlebectomy probe for periadventitial dissection and endoluminal probing during ambulatory phlebectomy. METHODS: The techniques of periadventitial dissection and endoluminal probing are reviewed. RESULTS: The phlebectomy probe seems to permit extraction of longer segments of varicose veins after releasing periadventitial adhesions. Endoluminal probing can assist in difficult localization of venous segments. CONCLUSION: The phlebectomy probe can be helpful for releasing periadventitial adhesions that tether varicose veins. Its application is simple and can result in increased length of avulsed varicose vein segments, thereby reducing the number of incisions needed. PMID- 10469118 TI - Multiple primary melanoma. AB - BACKGROUND: Incidence rates of cutaneous malignant melanoma (CMM) have been increasing for decades among Caucasian populations worldwide. Multiple factors identify persons at increased risk of CMM, including those with a family history of melanoma and those with atypical moles. Approximately 6-12% of melanomas are familial and approximately 12% of patients with familial melanoma have multiple primary melanomas. OBJECTIVE: To report a case of a patient with atypical moles and with 17 multiple primary melanomas. To review the literature on multiple primary melanomas as well as to review the genetics and treatment of melanoma. CONCLUSION: Patients with numerous atypical moles and a family or personal history of melanoma are at greatest risk for developing CMM. Patients from this population tend to develop CMM approximately 10 years earlier than the general population and have an increased risk for developing multiple primary melanomas. Since genetic tests capable of detecting individuals with an inherited susceptibility to CMM are not available, it is important to identify those patients with an increased risk and monitor them closely with regular total-body examinations. PMID- 10469119 TI - Electrosurgical skin resurfacing: a new bipolar instrument. AB - BACKGROUND: Numerous modalities may be used for skin resurfacing, including chemical peels, dermabrasion, and lasers. Each of these methods is associated with significant disadvantages. OBJECTIVE: The purpose of these initial studies was to determine the efficacy and safety of a new electrosurgical resurfacing system. Depth of cutaneous injury was also evaluated. METHODS: Postoperative scar resurfacing was performed on six patients in the initial feasibility study. Patients were evaluated with questionnaires, physician observations, and photographs. The histologic investigation evaluated depth of injury after resurfacing at various power settings and number of passes. RESULTS: Appearance of postoperative scars in all 6 patients was improved by electrosurgical resurfacing. The overall injury, residual thermal damage plus ablation, for all power levels and passes was 114.1 micrometer (mean) with a standard deviation of 60.7 micrometer. CONCLUSION: Electrosurgical resurfacing may become an effective and safe alternative to current resurfacing modalities. PMID- 10469120 TI - Invasive cutaneous squamous cell carcinoma associated with actinic keratosis: a case with orbital invasion and meningeal infiltration. AB - BACKGROUND: It is suggested that most squamous cell carcinomas in sun-exposed areas arise from preexisting solar keratosis. Actinic keratosis is thought of as being a precursor to squamous cell carcinoma. This form of squamous cell carcinoma has been considered to be a relatively benign lesion. We report a case of invasive squamous cell carcinoma associated with actinic keratosis leading to orbit destruction and meningeal infiltration. OBJECTIVE: To demonstrate that well differentiated tumors can act extremely aggressively with the potential toward infiltrative growth patterns. METHODS: Histologically controlled surgery along with multiple radiation therapy was performed. RESULTS: The tumor progressed inducing perineural invasion, orbit infiltration, osseous destruction, and meningeal invasion. CONCLUSION: The association of squamous cell carcinoma and actinic keratosis supports the concept of a causal relation. Excision with histologic examination of actinic keratosis seems to be useful for accurate diagnosis. Squamous cell carcinoma can represent an aggressive tumor with infiltrative growth pattern and should not be considered a benign lesion. PMID- 10469122 TI - Reticular veins and telangiectasia: a personal commentary. PMID- 10469121 TI - Chemical injury to the eye from EMLA cream during erbium laser resurfacing. AB - BACKGROUND: EMLA cream is commonly used as a topical anesthetic by physicians performing dermatologic surgery. OBJECTIVE: The purpose of this article is to inform physicians that EMLA cream is highly alkaline and can be particularly toxic to the eye. Symptoms of eye irritation may be initially masked because of local anesthetic effects. METHODS: Two patients are described who developed corneal abrasions from inadvertent exposure of the eye to EMLA cream prior to erbium laser resurfacing. RESULTS: The patients developed corneal abrasions and conjunctivitis, consistent with chemical alkaline burns. CONCLUSION: EMLA cream should be used with extreme caution near the eye and probably should be avoided when there is a need to insert laser eye shields. PMID- 10469123 TI - PTEN/MMAC1/TEP1 in signal transduction and tumorigenesis. AB - The level of phosphorylation within cells is tightly regulated by the concerted action of protein kinases and protein phosphatases [Hunter, T. (1995) Cell 80, 225-236]. Disregulation in the activity of either of these players can lead to cellular transformation. Many protein tyrosine kinases are proto-oncogenes and it has been postulated that some protein phosphatases may act as tumor suppressors. Herein we will review the recent findings addressing the roles the candidate tumor suppressor PTEN/MMAC1/TEP1 (PTEN, phosphatase and tensin homologue deleted from chromosome 10; MMAC 1, mutated in multiple advanced cancers 1; TEP1, TGF beta regulated and epithelial cell enriched phosphatase 1) plays in signal transduction and tumorigenesis. PTEN is a dual specificity protein phosphatase (towards phospho-Ser/Thr and phospho-Tyr) and, unexpectedly, also has a phosphoinositide 3-phosphatase activity. PTEN plays an important role in the modulation of the 1-phosphatidylinositol 3-kinase (PtdIns 3-kinase) pathway, by catalyzing the degradation of the PtdIns(3,4,5)P3 generated by PtdIns 3-kinase; this inhibits the downstream functions mediated by the PtdIns 3-kinase pathway, such as activation of protein kinase B (PKB, also known as Akt), cell survival and cell proliferation. Furthermore, PTEN modulates cell migration and invasion by negatively regulating the signals generated at the focal adhesions, through the direct dephosphorylation and inhibition of focal adhesion kinase (FAK). Growth factor receptor signaling is also negatively regulated by PTEN, through the inhibition of the adaptor protein Shc. While some of the functions of PTEN have been elucidated, it is clear that there is much more to discover about the roles of this unique protein. PMID- 10469124 TI - Evidence that phospholipase C-gamma2 interacts with SLP-76, Syk, Lyn, LAT and the Fc receptor gamma-chain after stimulation of the collagen receptor glycoprotein VI in human platelets. AB - Platelet activation by collagen is mediated by the sequential tyrosine phosphorylation of the Fc receptor gamma-chain (FcR gamma-chain), which is part of the collagen receptor glycoprotein VI, the tyrosine kinase Syk and phospholipase C-gamma2 (PLC-gamma2). In this study tyrosine-phosphorylated proteins that associate with PLC-gamma2 after stimulation by a collagen-related peptide (CRP) were characterized using glutathione S-transferase fusion proteins of PLC-gamma2 Src homology (SH) domains and by immunoprecipitation of endogenous PLC-gamma2. The majority of the tyrosine-phosphorylated proteins that associate with PLC-gamma2 bind to its C-terminal SH2 domain. These were found to include PLC-gamma2, Syk, SH2-domain-containing leucocyte protein of 76 kDa (SLP-76), Lyn, linker for activation of T cells (LAT) and the FcR gamma-chain. Direct association was detected between PLC-gamma2 and SLP-76, and between PLC-gamma2 and LAT upon CRP stimulation of platelets by far-Western blotting. FcR gamma chain and Lyn were found to co-immunoprecipitate with PLC-gamma2 as well as with unidentified 110-kDa and 75-kDa phosphoproteins. The absence of an in vivo association between Syk and PLC-gamma2 in platelets is in contrast with that for PLC-gamma1 and Syk in B cells. The in vivo function of PLC-gamma2 SH2 domains was examined through measurement of Ca2+ increases in mouse megakaryocytes that had been microinjected with recombinant proteins. This revealed that the C-terminal SH2 domain is involved in the regulation of PLC-gamma2. These data indicate that the C-terminal SH2 domain of PLC-gamma2 is important for PLC-gamma2 regulation through possible interactions with SLP-76, Syk, Lyn, LAT and the FcR gamma-chain. PMID- 10469125 TI - Presence of asparagine-linked N-acetylglucosamine and chitobiose in Pyrus pyrifolia S-RNases associated with gametophytic self-incompatibility. AB - S-RNases encoded by the S-locus of rosaceous and solanaceous plants discriminate between the S-alleles of pollen in gametophytic self-incompatibility reactions, but it is not clear how. We report the structures of N-glycans attached to each of the N-glycosylation sites of seven S-RNases in Pyrus pyrifolia of the Rosaceae. The structures were identified by chromatographic analysis of pyridylaminated sugar chains prepared from S4-RNase and by liquid chromatography/electrospray ionization-mass spectrometric analysis of the protease digests of reduced and S-carboxymethylated S-RNases. S4-RNase carries various types of sugar chains, including plant-specific ones with beta1-->2 linked xylose and alpha1-->3-linked fucose residues. More than 70% of the total N glycans of S4-RNase are, however, an N-acetylglucosamine or a chitobiose (GlcNAcbeta1-->4GlcNAc), which has not been found naturally. The N acetylglucosamine and chitobiose are mainly present at the N-glycosylation sites within the putative recognition sites of the S-RNase, suggesting that these sugar chains may interact with pollen S-product(s). PMID- 10469126 TI - Refinement and evaluation of a model of Mg2+ buffering in human red cells. AB - The total Mg2+ content of human red cells ([Mg]T,i) is partitioned between free and bound forms. The main cytoplasmic Mg2+ buffers are ATP and 2,3 bisphosphoglycerate. Haemoglobin binds free ATP and bisphosphoglycerate, preferentially in the deoxygenated state. Thus, the free ionized Mg2+ concentration ([Mg2+]i) oscillates with the oxy-deoxy condition of the cells. The binding reactions are also modulated by the pH changes that accompany the oxygenation-deoxygenation transitions. The complex interactions between Mg2+, its ligands and Hb can be encoded in a set of equilibrium equations representing all the known binding reactions of the system. To develop a comprehensive understanding of the Mg2+ homeostasis of intact red cells it is necessary to correct and refine the equations and parameters of the model by systematic comparisons between model predictions and measured cytoplasmic Mg2+ buffering curves under a variety of experimental conditions. Earlier models largely underestimated total Mg2+ binding in intact cells. We carried out experiments in which [Mg]T,i and [Mg2+]i were controlled over a wide range ([Mg]T,i between 0.1 and 23 mM) by the use of the ionophore A23187, under diverse metabolic conditions, and the results were used to interpret the adjustments required for good model fits. By the inclusion of low-affinity Mg2+ binding to ATP and bisphosphoglycerate, and also binding of Mg2+ to haemoglobin (four ions per tetramer) with an apparent dissociation constant of 45 mM we were able to realistically model, for the first time, all the experimentally observed changes in [Mg2+]i in human red cells under diverse metabolic conditions. PMID- 10469127 TI - Binding of AR-1-144, a tri-imidazole DNA minor groove binder, to CCGG sequence analyzed by NMR spectroscopy. AB - The interactions of N-[2-(dimethylamino)ethyl]-1-methyl-4-[1-methyl-4-[4 formamido-1-meth ylimidazole-2-carboxamido]imidazole-2-carboxamido]imidazole-2-c arboxa mide (AR-1-144), a tri-imidazole polyamide minor groove binder, with DNA have been investigated by NMR and CD spectroscopy. A series of DNA oligonucleotides with a C/G-containing four-bp core, i.e. CCGG, CGCG, GGCC, and GCGC, have been titrated with AR-1-144 at different ratios. AR-1-144 favors the CCGG sequence. The flanking sequence of the CCGG core also influences the binding preference, with a C or T being favored on the 3'-side of the CCGG core. The three-dimensional structure of the symmetric 2:1 side-by-side complex of AR-1-144 and GAACCGGTTC, determined by NOE-constrained NMR refinement, reveals that each AR-1-144 binds to four base pairs, i.e. at C5-G6-G7-T8, with every amide imidazole unit forming two potential hydrogen bonds with DNA. The same DNA binding preference of AR-1-144 was also confirmed by circular dichroism spectroscopy, indicating that the DNA binding preference of AR-1-144 is independent of concentration. The cooperative binding of an AR-1-144 homodimer to the (purine)CCGG(pyrimidine) core sequence appears to be weaker than that of the distamycin A homodimer to A/T sequences, most likely due to the diminished hydrophobic interactions between AR-1-144 and DNA. Our results are consistent with previous footprinting data and explain the binding pattern found in the crystal structure of a di-imidazole drug bound to CATGGCCATG. PMID- 10469128 TI - Structure determination of the O-antigenic polysaccharide from the enteroinvasive Escherichia coli O136. AB - The structure of the O-antigen polysaccharide of the lipopolysaccharide from the enteroinvasive Escherichia coli O136 has been elucidated. The composition of the repeating unit was established by sugar and methylation analysis together with 1H and 13C NMR spectroscopy. Two-dimensional nuclear Overhauser effect spectroscopy (NOESY) and heteronuclear multiple-bond correlation experiments were used to deduce the sequence. The absolute configuration for the nonulosonic acid (NonA) could be determined using spin-spin coupling constants, 13C chemical shifts and NOESY. The anomeric configuration of the NonA was determined via vicinal and geminal 13C,1H coupling constants. The structure of the repeating unit of the polysaccharide from E. coli O136 is as follows, in which beta-NonpA is 5,7 diacetamido-3,5,7, 9-tetradeoxy-Lglycero-beta-Lmanno-nonulosonic acid: -->4)-beta NonpA-(2-->4)-beta-D-Galp-(1-->4)-beta-D-GlcpNAc-(1--> PMID- 10469129 TI - Nitrile hydratase involved in aldoxime metabolism from Rhodococcus sp. strain YH3 3 purification and characterization. AB - Nitrile hydratase responsible for aldoxime metabolism from the E-pyridine-3 aldoxime degrading bacterium, Rhodococcus sp. strain YH3-3 was purified and characterized. Addition of cobalt ion was necessary for the formation of enzyme. The enzyme activity was highly induced not only by nitriles and amides but also by several aldoxime compounds. The enzyme was purified approximately 108-fold with a 16% yield from the cell-free extract of the strain. The native enzyme had a Mr of approximately 130 000 and consisted of two subunits (alpha-subunit, 27 100; beta-subunit, 34 500). The enzyme contained approximately 2 mol cobalt per mol enzyme; it showed a maximum activity at 60 degrees C and at 40 degrees C under the rate assay and end-point assay conditions, respectively, and was stable over a wide range of pH (pH 2.5-11.0). The enzyme had a wide substrate specificity: it acted on aliphatic saturated and unsaturated as well as aromatic nitriles. The N-terminus of the beta-subunit showed good sequence similarities with those of other nitrile hydratases. Nitrile hydratase is part of the metabolic pathway for aldoximes in microorganisms. PMID- 10469130 TI - Top-down control analysis of ATP turnover, glycolysis and oxidative phosphorylation in rat hepatocytes. AB - Control analysis was used to analyse the internal control of rat hepatocyte metabolism. The reactions of the cell were grouped into nine metabolic blocks linked by five key intermediates. The blocks were glycogen breakdown, glucose release, glycolysis, lactate production, NADH oxidation, pyruvate oxidation, mitochondrial proton leak, mitochondrial phosphorylation and ATP consumption. The linking intermediates were intracellular glucose-6-phosphate, pyruvate and ATP levels, cytoplasmic NADH/NAD ratio and mitochondrial membrane potential. The steady-state fluxes through the blocks and the levels of the intermediates were measured in the absence and presence of specific effectors of hepatocyte metabolism. Application of the multiple modulation approach gave the kinetic responses of each block to each intermediate (the elasticities). These were then used to calculate all of the control coefficients, which describe the degree of control each block had over the level of each intermediate, and over the rate of each process. Within this full description of control, many different interactions could be identified. One key finding was that the processes that consumed ATP had only 35% of the control over the rate of ATP consumption. Instead, the reactions that produced ATP exerted the most control over ATP consumption rate; particularly important were mitochondrial phosphorylation (30% of control) and glycolysis (19%). The rate of glycolysis was positively controlled by the glycolytic enzymes themselves (66% of control) and by ATP consumption (47%). Mitochondrial production of ATP, including oxidative, proton leak and phosphorylation processes, had negative control over glycolysis (-26%; the Pasteur effect). In contrast, glycolysis had little control over the rate of ATP production by the mitochondria (-10%; the Crabtree effect). Control over the flux through the mitochondrial phosphorylation block was shared between pyruvate oxidation (23%), ATP consumption (28%) and the mitochondrial phosphorylation block itself (64%). PMID- 10469131 TI - Identification and characterization of cDNA clones encoding hydroxycinnamoyl CoA:tyramine N-hydroxycinnamoyltransferase from tobacco. AB - The sequences of three cDNA clones that include the complete coding region of hydroxycinnamoyl-CoA:tyramine N-hydroxycinnamoyltransferase (THT) from tobacco are reported. The three cDNAs were isolated by antibody screening of a cDNA expression library produced from poly(A)+RNA purified from tobacco leaves (Nicotiana tabacum cv. Bottom Special), previously infiltrated with an incompatible strain of Ralstonia solanacearum. The identity of these clones was confirmed by the detection of THT activity in extracts of transformed Escherichia coli and by matching the translated polypeptides with tryptic enzyme sequences. cDNA clones tht4 and tht11 differ only by their 5' leader and 3' UTRs and therefore encode the same protein, whereas tht10 and tht11 exhibit 95 and 99% sequence identity at the DNA and deduced amino acid levels, respectively. The three clones encode proteins of 226 amino acids with calculated molecular masses of 26 kDa. The deduced amino acid sequences show no similarity with the sequence of anthranilate hydroxycinnamoyl/benzoyltransferase from Dianthus caryophyllus, the only enzyme exhibiting hydroxycinnamoyltransferase activity to be cloned so far in plants. In contrast, comparison of the THT amino acid sequence with protein sequence databases revealed substantial homology with mammalian diamine acetyltransferases. The THT clones hybridized to a 0.95-kb mRNA from elicited tobacco cell-suspension cultures and also to a mRNA of similar size from wound healing potato tubers. The messengers for THT were also found to be expressed at relatively high levels in tobacco root tissues. Southern hybridization of tobacco genomic DNA with THT cDNA suggests that several copies of the THT gene occur in the tobacco genome. Inhibition experiments using amino-acid-specific reagents demonstrated that both histidyl and cysteyl residues are required for THT activity. In the course of these experiments THT was also found to be inhibited by (2-hydroxyphenyl) amino sulfinyl acetic acid 1,1-dimethylethyl ester, an irreversible inhibitor of cinnamyl alcohol dehydrogenase. PMID- 10469132 TI - 31P and 13C nuclear magnetic resonance studies of metabolic pathways in Pasteurella multocida characterization of a new mannitol-producing metabolic pathway. AB - Glucose metabolism of Pasteurella multocida was examined in resting cells in vivo using 13C NMR spectroscopy, in cell-free extracts in vitro using 31P NMR spectroscopy and using enzyme assays. The NMR data indicate that glucose is converted by the Embden-Meyerhof and pentose phosphate pathways. The P. multocida fructose 6-phosphate phosphotransferase activity (the key enzyme of the Embden Meyerhof pathway) was similar to that of Escherichia coli. Nevertheless, and in contrast to that of E. coli, its activity was inhibited by alpha glycerophosphate. This inhibition is consistent with the very low fructose 6 phosphate phosphotransferase activity found in cell-free extracts of P. multocida using a spectrophotometric method. The dominant end products of glucose metabolism were mannitol, acetate and succinate. Under anaerobic conditions, P. multocida was able to constitutively produce mannitol from glucose, mannose, fructose, sucrose, glucose 6-phosphate and fructose 6-phosphate. We propose a new metabolic pathway in P. multocida where fructose 6-phosphate is reduced to mannitol 1-phosphate by fructose 6-phosphate reductase. Mannitol 1-phosphate produced is then converted to mannitol by mannitol 1-phosphatase. PMID- 10469133 TI - Cyclic AMP-dependent synthesis and release of adrenomedullin and proadrenomedullin N-terminal 20 peptide in cultured bovine adrenal chromaffin cells. AB - Adrenomedullin and proadrenomedullin N-terminal 20 peptide are peptides with multiple physiological functions and are most abundant in adrenal medulla. We studied whether the cAMP-dependent pathway is involved in the regulation of synthesis and release of adrenomedullin and proadrenomedullin N-terminal 20 peptide in cultured bovine adrenal chromaffin cells. Exposure of the cells to dibutyryl cAMP (dbcAMP) increased a progressive accumulation of immunoreactive adrenomedullin and immunoreactive-proadrenomedullin N-terminal 20 peptide in the extracellular medium, while reciprocally decreasing their cellular content in a time-dependent manner. The decrease of levels of both peptides in the cells was much greater in extent than the increase of the peptides in the medium. H89, an inhibitor of cAMP-dependent protein kinase attenuated these changes, induced by dbcAMP. The resulting changes by dbcAMP and H89 were similar to those of chromogranin B, a marker peptide of chromaffin granule. Northern blot analysis showed that the mRNA encoding these peptides, detected as a band of 1.6 kb, was decreased by the treatment with dbcAMP. The effect of dbcAMP on mRNA was attenuated by H89, and was reversible as the decreased mRNA level caused by dbcAMP could be returned to control levels by culturing cells after removal of dbcAMP. These results suggest that the cAMP-dependent protein kinase pathway stimulates the release of adrenomedullin and proadrenomedullin N-terminal 20 peptide, whereas it lowers synthesis of these peptides via the reduction of their transcript level. PMID- 10469134 TI - Selective targeting of an antioxidant to mitochondria. AB - Mitochondrial oxidative damage contributes significantly to a range of human disorders, including neurodegenerative diseases, ischaemia-reperfusion injury and ageing-associated dysfunction. To prevent this damage we have delivered a molecule containing the active antioxidant moiety of vitamin E to mitochondria. This was carried out by covalently coupling the antioxidant moiety to a lipophilic triphenylphosphonium cation. This mitochondrially targeted antioxidant, 2-[2-(triphenylphosphonio)ethyl]-3,4-dihydro-2, 5,7,8-tetramethyl-2H 1-benzopyran-6-ol bromide (TPPB), accumulated several-hundred fold within the mitochondrial matrix, driven by the organelle's large membrane potential. When cells were incubated with micromolar concentrations of TPPB, they accumulated millimolar concentrations within their mitochondria. The amount of TPPB taken up by mitochondria was approximately 80-fold greater than endogenous levels of vitamin E. Consequently the targeted derivative of vitamin E protected mitochondrial function from oxidative damage far more effectively than vitamin E itself. The mitochondrially targeted antioxidant TPPB has potential as an antioxidant therapy for disorders involving mitochondrial oxidative damage. It also suggests a new family of mitochondrially targeted antioxidants, redox-active and pharmacologically active molecules designed to prevent damage or manipulate mitochondrial function. PMID- 10469135 TI - The half-life of human procathepsin S. AB - Two processes, synthesis and degradation, contribute to the intracellular concentration of a protein. As most malignant tumors or tumor cell lines show elevated levels of proteinases, we studied the half-life of a cysteine proteinase, procathepsin S, in order to determine whether tumor cells can regulate their cathepsin concentration via changing the degradation rate of the enzyme. The following procathepsin S species were examined: wild-type procathepsin S in macrophages, recombinant procathepsin S in human embryonic kidney cells (HEK 293 cells), recombinant nonglycosylated procathepsin S in HEK 293 cells, wild-type procathepsin S in the established nonsmall cell lung carcinoma cell line 97TM1. The half-lives of both wild-type procathepsins S expressed in macrophages and in HEK 293 cells were 1 h, whereas that of procathepsin S in the tumor cell line was 2 h. Nonglycosylated procathepsin S was not processed. The degradation of mature cathepsin S proceeded with a half-life of 16-18 h. All cell lines studied secreted substantial amounts of procathepsin S into the culture medium. No further maturation of secreted procathepsin S has been observed in the culture medium. We suggest a disturbed sorting mechanism in tumor cells. PMID- 10469136 TI - Overexpression, purification and biochemical characterization of the wound induced leucine aminopeptidase of tomato. AB - Wounding of tomato leaves results in the accumulation of an exoprotease called leucine aminopeptidase (LAP-A). While the expression of LapA genes are well characterized, the specificity of the LAP-A enzyme has not been studied. The LAP A preprotein and mature polypeptide were overexpressed in Escherichia coli. PreLAP-A was not processed and was inactive accumulating in inclusion bodies. In contrast, 55-kDa mature LAP-A subunits assembled into an active, 357-kDa enzyme in E. coli. LAP-A from E. coli cultures was purified to apparent homogeneity and characterized relative to its animal (porcine LAP) and prokaryotic (E. coli PepA) homologues. Similar to the porcine and E. coli enzymes, the tomato LAP-A had high temperature and pH optima. Mn2+ was a strong activator for all three enzymes, while chelators, zinc ion, and the slow-binding aminopeptidase inhibitors (amastatin and bestatin) strongly inhibited activities of all three LAPs. The substrate specificities of porcine, E. coli and tomato LAPs were determined using amino-acid-p-nitroanilide and -beta-naphthylamide substrates. The tomato LAP-A preferentially hydrolyzed substrates with N-terminal Leu, Met and Arg residues. LAP-A had substantially lower levels of activity on other chromogenic substrates. Several differences in substrate specificities for the animal, plant and prokaryotic enzymes were noted. PMID- 10469137 TI - Alternative promoters direct tissue-specific expression of the mouse protein phosphatase 2Cbeta gene. AB - Type 2C protein phosphatases (PP2Cs), a class of ubiquitous and evolutionally conserved serine/threonine protein phosphatases, are encoded in at least four distinct genes and implicated in the regulation of various cellular functions. Of these four PP2C genes, the expression of the PP2Cbeta gene has been reported to be tissue-specific and development-dependent. To understand more precisely the regulatory mechanism of this expression, we have isolated and characterized overlapping mouse genomic lambda clones. A comparison of genomic sequences with PP2Cbeta cDNA sequences provided information on the structure and localization of intron/exon boundaries and indicated that PP2Cbeta isoforms with different 5' termini were generated by alternative splicing of its pre-mRNA. The 5'-flanking region of exon 1 had features characteristic of a housekeeping gene: it was GC rich, lacked TATA boxes and CAAT boxes in the standard positions, and contained potential binding sites for the transcription factor SP1. In the 5'-flanking region of exon 2, several consensus sequences were found, such as a TATA-like sequence and negative regulatory element box-1, -2 and -3. Subsequent analysis by transient transfection assay with a reporter gene showed that these regions act as distinct promoters. Analysis of PP2Cbeta transcripts by reverse transcriptase PCR showed that exon-1 transcripts were expressed ubiquitously in all of the tissues examined, whereas exon-2 transcripts were predominantly expressed in the testis, intestine and liver. These results suggest that the alternative usage of two promoters within the PP2Cbeta gene regulates tissue-specific expression of PP2Cbeta mRNA. PMID- 10469138 TI - Human chemokine receptors CCR5, CCR3 and CCR2B share common polarity motif in the first extracellular loop with other human G-protein coupled receptors implications for HIV-1 coreceptor function. AB - Chemokine receptors (CRs) are 7-helix membrane proteins from the family of G protein coupled receptors (GPCRs). A few human CRs act as cofactors for macrophage-tropic (M-tropic) human immunodeficiency virus type-1 (HIV-1) entry into cells, while others do not. In this study, we describe an application of molecular modeling techniques to delineate common molecular determinants that might be related to coreceptor activity, and the use of the data to identify other GPCRs as putative cofactors for M-tropic HIV-1 entry. Subsequently, the results were confirmed by an experimental approach. The sequences of extracellular domains (ECDs) of CRs were employed in a compatibility search against a database of environmental profiles derived for proteins with known spatial structure. The best-scoring sequence-profile alignments obtained for each ECD were compared in pairs to check for common patterns in residue environments, and consensus sequence-profile fits for ECDs were also derived. Similar hydrophobicity motifs were found in the first extracellular loops of the CRs CCR5, CCR3, and CCR2B, and are all used by M-tropic HIV-1 for cell entry. In contrast, other CRs did not reveal common motifs. However, the same environmental pattern was also delineated in the first extracellular loop of some human GPCRs showing either high (group 1) or low (group 2) degree of similarity of their polarity patterns with those in HIV-1 coreceptors. To address the question of whether the delineated molecular determinant plays a critical role in the receptor-virus binding, three of the identified GPCRs, bradykinin receptor (BRB2) and G-protein receptor (GPR)-CY6 from group 1, and GPR8 from group 2, were cloned and transfected into HeLa-CD4 cells, which are nonpermissive to M-tropic HIV-1 infection. We demonstrate that, similar to CCR5, the two selected GPCRs from group 1 were capable of mediating M-tropic HIV-1 entry, whereas GPR8 from group 2 did not serve as HIV-1 coreceptor. The potential biological significance of the identified structural motif shared by the human CCR5, CCR3, CCR2B and other GPCRs is discussed. PMID- 10469139 TI - Down-regulation of endothelial cell growth inhibitors by enhanced MYCN oncogene expression in human neuroblastoma cells. AB - Recent evidence indicates that the genetic alterations of the multistage process of malignant transformation appear to activate tumor neovascularization by altering the balance between stimulators and inhibitors of angiogenesis. In the present study, we have attempted to define the effect of enhanced MYCN oncogene expression on the profile of endothelial cell growth modulators in neuroblastoma cells. We report here that conditioned medium of human neuroblastoma cells with normal MYCN expression contains three inhibitors of endothelial cell proliferation, which appear to be novel proteins as judged by their physicochemical, immunological and biological properties. All three inhibitors are diminished or become undetectable upon experimental increase of MYCN expression. Our results suggest that enhanced MYCN expression in human neuroblastoma cells alters the angiogenic balance by down-regulating endothelial cell growth inhibitors but leaving the expression of the stimulators unaffected. These data shed light on the molecular mechanisms linking the genetic changes of malignant transformation with initiation of tumor angiogenesis. Moreover, our observations might explain the poor prognosis of human neuroblastomas following MYCN oncogene amplification through initiation of angiogenesis and subsequent tumor growth and spread. PMID- 10469140 TI - Plasminogen activator inhibitor type-2 (PAI-2) gene transcription requires a novel NF-kappaB-like transcriptional regulatory motif. AB - Induction of human plasminogen activator inhibitor type-2 (PAI-2) gene transcription is the response of macrophages to inflammatory stimuli, such as the pleiotropic cytokine, tumour necrosis factor-alpha (TNFalpha). Here we have examined whether PAI-2 gene transcription in response to TNFalpha may be mediated through a regulatory pathway involving the transcription factor, NF-kappaB. We have tested the function of two potential NF-kappaB-like sites present in the PAI 2 proximal promoter for responsiveness to TNFalpha using chloramphenicol acetyl transferase reporter gene deletion and mutation analyses. While no evidence was found for TNFalpha regulation of the PAI-2 gene through either of these two sites, one of the NF-kappaB-like motifs, transcriptional regulatory motif (TRM), present at position -400 was found to be essential for constitutive PAI-2 transcription, as mutation of this motif abolished basal PAI-2 promoter activity in both monocyte-like U937 cells and HT1080 fibrosarcoma cells. Competition electrophoretic mobility shift assays identified four TRM-binding proteins present in U937, HT1080 and HeLa cell extracts, which bound to this motif but were not components of the NF-kappaB regulatory complex. Expression screening of a HeLa cell cDNA library using the -400 TRM as a probe identified two cDNAs encoding partial peptides which specifically bound the TRM motif. DNA sequence analysis revealed that one cDNA was novel, and the second cDNA encoded exon 5 of the nephroblastoma overexpressed (novH) proto-oncogene, suggesting a new role for this peptide in gene regulation. Taken together, these findings identify a new regulatory element required for constitutive PAI-2 transcription, and identify potential DNA-binding proteins associated with this element that may play a role in PAI-2 gene regulation. PMID- 10469141 TI - Purified apolipoprotein B gene regulatory factor-3 is DNA topoisomerase I. AB - Hepatic cell-specific expression of the human apolipoprotein B (apoB) gene is controlled by at least four cis-acting elements located between positions -128 and +122 [Chuang, S. S., & Das, H. K. (1996) Biochem. Biophys. Res. Commun. 220, 553-562]. A negative cis-acting element (+20 to +40) is located in the first nontranslated exon of the human apoB gene, and apoB regulatory factor-3 (BRF-3) interacts with this. In this paper, we report the purification and characterization of BRF-3 from rat liver nuclear extracts. BRF-3 has been purified to apparent homogeneity by DEAE-cellulose, heparin-agarose, and DNA specific affinity chromatography. Purified BRF-3 produced two polypeptide bands with apparent molecular masses of 70 kDa and 67 kDa in SDS/PAGE as detected by silver staining. Both 70-kDa and 67-kDa proteins have been found to hybridize specifically with labeled double-stranded oligonucleotide containing BRF-3 binding site in a South-Western blot. Double-stranded oligonucleotide containing mutations in the BRF-3 binding site was found to abolish DNA binding by these two proteins. Amino acid sequences of tryptic peptides derived from affinity purified 70-kDa and 67-kDa rat BRF-3 proteins were found to have 100% sequence homologies with DNA topoisomerase I. These data suggest that the 70-kDa and 67-kDa forms of BRF-3 are derived by proteolytic cleavage of topoisomerase I, and therefore, topoisomerase I may play an important role in transcriptional regulation of apoB. PMID- 10469142 TI - Introduction of a C-terminal aromatic sequence from snake venom phospholipases A2 into the porcine pancreatic isozyme dramatically changes the interfacial kinetics. AB - Porcine pancreatic phospholipase A2 (PLA2) was modified by single and multiple site-directed mutations at sites thought to be involved in interfacial binding. Charged and polar residues in the C-terminal region were replaced by aromatic residues on the basis of an analogy with snake venom PLA2s, which display high affinity for a zwitterionic interface. The PLA2 variants constructed were N117W, N117W/D119Y and K116Y/N117W/D119Y. Titration with micelles of a zwitterionic substrate suggests that the variants N117W and K116Y/N117W/D119Y possess improved ability to bind to the micellar substrate interface, relative to the wild-type enzyme. Improved interfacial binding was confirmed by direct binding studies with micelles of a zwitterionic substrate analogue, indicating up to five times higher affinity for both variants. Interfacial binding is not improved for the variant N117W/D119Y. Maximal enzyme velocities (Vapp./max) with the zwitterionic substrate were between 25 and 75% of that of the wild-type enzyme. However, competitive inhibition and direct binding studies with a strong inhibitor revealed that the affinity for substrate present at the interface (Km*) is perturbed by the mutations made. For the variant N117W, the slight decrease observed in Vapp./max is most likely made up of a 24-fold reduction in catalytic turnover (kcat) and 18-fold improved substrate binding (Km*). PMID- 10469143 TI - Methylcobalamin:homocysteine methyltransferase from Methanobacterium thermoautotrophicum. Identification as the metE gene product. AB - Methanobacterium thermoautotrophicum is a methane-forming archaeon that grows on H2 and CO2 as sole carbon and energy source. Cell extracts of the methanogen were found to contain methylcobalamin: homocysteine methyltransferase activity which was purified 3000-fold to a specific activity of approximately 500 U.mg-1 protein. SDS/PAGE revealed the presence of a polypeptide with an apparent molecular mass of 34 kDa. Via its N-terminal amino acid sequence, the 34-kDa polypeptide was identified as the metE gene product. The metE gene was heterologously expressed in Escherichia coli. The overproduced protein was recovered in the inclusion body fraction and was found to be inactive. The protein could be partially solubilized by unfolding in 8 M urea and then refolding. The solubilized protein had a specific activity of 450 U.mg-1. It exhibited first-order kinetics with respect to methylcobalamin concentration and Michaelis-Menten kinetics with respect to L-homocysteine concentration (apparent Km 0.1 mM). The enzyme was specific for L-homocysteine as methyl acceptor. Methylcobalamin could be substituted with methylcobinamide as methyl donor. PMID- 10469144 TI - Developmental expression of the protein kinase C substrate/binding protein (clone 72/SSeCKS) in rat testis identification as a scaffolding protein containing an A kinase-anchoring domain which is expressed during late-stage spermatogenesis. AB - The coordinated interaction of kinases, phosphatases and other regulatory molecules with scaffolding proteins is emerging as a major theme in intracellular signaling networks. In this report we show that a cDNA isolated from a rat testis expression library by interactive cloning using the regulatory subunit (R) of a type-II protein kinase A (PKA) is identical with a previously characterized protein kinase C (PKC)-binding protein termed either clone 72 [Chapline, C., Mousseau, B., Ramsay, K., Duddy, S., Li, Y., Kiley, S. C. & Jaken, S. (1996) J. Biol. Chem. 271, 6417-6422] or SSeCKS [Lin, X., Tombler, E., B., Nelson, P.J., Ross, M. & Gelman, I.H. (1996) J. Biol. Chem. 271, 28430-28438]. Deletion mutagenesis demonstrated that amino acids 1495-1524 of clone 72/SSeCKS had the ability to interact with RII. Antibodies prepared against the recombinant protein recognized a 280/290-kDa doublet and a 240-kDa protein on Western blots of rat testis cytosolic and Triton X-100 extracts. Expression of clone 72/SSeCKS mRNA and protein levels was developmentally regulated in rat testis. Northern-blot analysis showed a dramatic increase in clone 72/SSeCKS-hybridizing mRNA starting 30 days after birth. Immunohistochemical examination showed high expression levels in elongating spermatids. Clone 72/SSeCKS was not detected in mature sperm. These studies suggest a role for clone 72/SSeCKS, a PKA/PKC scaffolding protein, during the process of spermiogenesis. PMID- 10469145 TI - p55-hGRF, a short natural form of the Ras-GDP exchange factor high yield production and characterization. AB - p55-hGRF, a natural short form of the guanine-nucleotide-releasing factor for p21 Ras from human brain, was expressed at high level in Escherichia coli as well as an engineered truncated form, p39-hGRF. A T7 polymerase expression system was used, resulting in the formation of insoluble cytoplasmic protein aggregates. The recombinant products were resolubilized, renatured and purified to homogeneity. The exchange activity of the refolded hGRF samples on H-Ras was comparable with that published for the soluble catalytic domain of the mouse counterpart, CDC25 Mm. Both p55-hGRF and p39-hGRF form dimers. We established a procedure to prepare and purify the complex with Ras. The results of the characterization study are consistent with a stoichiometry of 1:1 and an equilibrium between dimeric and monomeric forms of the complex. PMID- 10469146 TI - Stabilization of DNA triple helices by a series of mono- and disubstituted amidoanthraquinones. AB - We have used quantitative DNase I footprinting to measure the relative affinities of four disubstituted and two monosubstituted amidoanthraquinone compounds for intermolecular DNA triplexes, and have examined how the position of the attached base-functionalized substituents affects their ability to stabilize DNA triplexes. All four isomeric disubstituted derivatives examined stabilize DNA triplexes at micromolar or lower concentrations. Of the compounds studied the 2,7 disubstituted amidoanthraquinone displayed the greatest triplex affinity. The order of triplex affinity for the other disubstituted ligands decreases in the order 2,7 > 1,8 = 1,5 > 2,6, with the equivalent monosubstituted compounds being at least an order of magnitude less efficient. The 1,5-disubstituted derivative also shows some interaction with duplex DNA. These results have been confirmed by molecular modelling studies, which provide a rational basis for the structure activity relationships. These suggest that, although all of the compounds bind through an intercalative mode, the 2,6, 2,7 and 1,5 disubstituted isomers bind with their two side groups occupying adjacent triplex grooves, in contrast with the 1,8 isomer which is positioned with both side groups in the same triplex groove. PMID- 10469147 TI - Regulation of type V phospholipase A2 expression and function by proinflammatory stimuli. AB - Types IIA and V secretory phospholipase A2 (sPLA2) are structurally related to each other and their genes are tightly linked to the same chromosome locus. An emerging body of evidence suggests that sPLA2-IIA plays an augmentative role in long-term prostaglandin (PG) generation in cells activated by proinflammatory stimuli; however, the mechanism underlying the functional regulation of sPLA2-V remains largely unknown. Here we show that sPLA2-V is more widely expressed than sPLA2-IIA in the mouse, in which its expression is elevated by proinflammatory stimuli such as lipopolysaccharide. In contrast, proinflammatory stimuli induced sPLA2-IIA in marked preference to sPLA2-V in the rat. Cotransfection of sPLA2-V with cyclooxygenase (COX)-2, but not with COX-1, into human embryonic kidney 293 cells dramatically increased the interleukin-1-dependent PGE2 generation occurring over a 24 h of culture period. Rat mastocytoma RBL-2H3 cells overexpressing sPLA2-V exhibited increased IgE-dependent PGD2 generation and accelerated beta-hexosaminidase exocytosis. These results suggest that sPLA2-V acts as a regulator of inflammation-associated cellular responses. This possible compensation of sPLA2-V for sPLA2-IIA in many, if not all, tissues may also explain why some mouse strains with natural disruption of the sPLA2-IIA gene exhibit few abnormalities during their life-spans. PMID- 10469148 TI - p-Coumaroyltriacetic acid synthase, a new homologue of chalcone synthase, from Hydrangea macrophylla var. thunbergii. AB - Chalcone synthase and stilbene synthase are plant-specific polyketide synthases. They catalyze three common consecutive decarboxylative condensations and specific cyclization reactions. They are highly homologous to each other, and are likely to fall into a family of polyketide synthases along with acridone synthase and bibenzyl synthase. Two cDNA clones (named HmC and HmS), both of which show high homology to the known chalcone synthases, were obtained from leaves of Hydrangea macrophylla var. thunbergii. They were expressed in Escherichia coli in order to determine their enzyme functions. Detection of chalcone formation clearly indicated that HmC encoded chalcone synthase, while HmS protein catalyzed the formation of neither chalcone nor stilbene. However, a novel pyrone, a lactonization product of a linear tetraketide was detected in reaction products of HmS protein. This proves that HmS encodes a novel polyketide synthase that catalyzes only chain elongation without cyclization. PMID- 10469149 TI - Cell adhesion to tenascin-X mapping of cell adhesion sites and identification of integrin receptors. AB - Adhesive properties of tenascin-X (TN-X) were investigated using TN-X purified from bovine skin and recombinant proteins encompassing the RGD sequence located within the tenth fibronectin type-III domain, and the fibrinogen-like domain. Osteosarcoma (MG63) and bladder carcinoma cells (ECV304) cells were shown to adhere to purified TN-X, but did not spread and did not assemble actin stress fibers. Both cell types adhered to recombinant proteins harboring the contiguous fibronectin type-III domains 9 and 10 (FNX 9-10) but not to the FNX 10 domain alone. This adhesion to FNX 9-10 was shown to be mediated by alphavbeta3 integrin, was inhibited by RGD peptides and was strongly reduced in proteins mutated within the RGD site. As antibodies against alphavbeta3 integrin had no effects on cell adhesion to purified TN-X, we suggest that the RGD sequence is masked in intact TN-X. Cell attachment to the recombinant TN-X fibrinogen domain (FbgX) and to purified TN-X was greater for MG63 than for ECV304 cells. A beta1 containing integrin was shown to be involved in MG63 cell attachment to FbgX and to purified TN-X. Although the existence of other cell interaction sites is likely in this huge molecule, these similar patterns of adhesion and inhibition suggest that the fibrinogen domain might be a dominant site in the whole molecule. PMID- 10469150 TI - The carbohydrates of the isoforms of three avian riboflavin-binding proteins. AB - The carbohydrate chains of nine isoforms of chicken egg-white riboflavin-binding protein (RfBP) and six isoforms each of quail egg-white and yolk RfBP have been structurally characterized. The two N-glycosylation sites, Asn36 and Asn147, of the most abundant isoform of each of the three proteins were analyzed in further detail leading to the identification of different glycosylation patterns. In both chicken and quail egg-white RfBP the carbohydrates attached to position 36 had a lower degree of branching and, in the case of the quail protein, this site was only partially glycosylated. A very heterogeneous mixture of complex structures was characteristic of the other glycosylation site. Analysis of the two sites in quail yolk RfBP confirmed this result which agrees with what has been established for hen yolk RfBP. The presence in the three proteins of a highly heterogeneous mixture of differently branched glycans suggests that the differences in isoelectric points, which is a peculiarity of the different isoforms, are probably indeed due to differences in carbohydrate structure. PMID- 10469151 TI - Insulin stimulates triacylglycerol secretion by perfused livers from fed rats but inhibits it in livers from fasted or insulin-deficient rats implications for the relationship between hyperinsulinaemia and hypertriglyceridaemia. AB - We determined whether the direction of the acute effect of insulin on hepatic triacylglycerol secretion is dependent on the prior physiological state or on the in vitro experimental system used. The effect of insulin on triacylglycerol secretion was studied using perfused livers isolated from rats under three metabolic conditions: fed normo-insulinaemic, 24-h fasted and fed, streptozotocin diabetic (insulin-deficient). Insulin acutely activated triacylglycerol secretion (by 43%) in organs from fed, normo-insulinaemic animals, whereas it inhibited triacylglycerol secretion in livers isolated from fasted or insulin-deficient rats (by 30 and 33%, respectively). By contrast, in 24-h-cultured hepatocytes insulin invariably acutely inhibited triacylglycerol secretion irrespective of the metabolic state of the donor animals. It is concluded that the use of perfused livers enables the observation of a switch in the direction of insulin action on hepatic triacylglycerol secretion from stimulatory, in the normo insulinaemic state, to inhibitory in the fasting or insulin-deficient state. The possible implications of this switch for the relationship between hyperinsulinaemia, increased hepatic very-low-density lipoprotein-triacylglycerol secretion and hypertriglyceridaemia observed in vivo are discussed. PMID- 10469152 TI - Ectoplasmic insertion of a glycosylphosphatidylinositol-anchored protein in glycosphingolipid- and cholesterol-containing phosphatidylcholine vesicles. AB - Glycosylphosphatidylinositol (glycosyl-PtdIns)-anchored proteins are proposed to be clustered in membrane microdomains enriched in cholesterol and glycosphingolipids (GlySphs). We have prepared biomimetic membranes in order to study the possible phenomena of surface aggregation of these membrane components. Phosphatidylcholine liposomes were treated by octylglucoside to insert a glycosyl PtdIns-protein, alkaline phosphatase (ALP), some cholesterol, and a GlySph, the lactocerebroside. The association of these compounds was shown by centrifugation on a density gradient. The presence of ALP on the surface of the vesicles was shown by the action of a phospholipase, and the presence of the lactocerebroside was shown by the use of a galactose-specific tetravalent lectin. Our data show that total alkaline phosphatase and half to total lactocerebroside were ectoplasmically inserted in the vesicles membrane. In addition, we observed that the presence of small amounts of ALP in the liposomes led to significant changes in membrane stability with regard to detergent, as shown by the changes in the solubilization process monitored by turbidimetry. Furthermore, we have built an original method to study the cohesion of the vesicles membrane, in which some magnesium ions were trapped in the luminal space of the liposomes during several days. The ALP is magnesium-dependent for its catalytic activity and was inhibited after incubation of ALP-containing liposomes in a magnesium-free buffer. The ALP activity was restored by the addition of detergent to the liposomes, due to the release of the luminal magnesium ions. Surface aggregation phenomena will now be investigated by atomic force microscopy. PMID- 10469153 TI - Flavonol 2,4-dioxygenase from Aspergillus niger DSM 821, a type 2 CuII-containing glycoprotein. AB - Flavonol 2,4-dioxygenase, which catalyzes the cleavage of quercetin to carbon monoxide and 2-protocatechuoyl-phloroglucinol carboxylic acid, was purified from culture filtrate of Aspergillus niger DSM 821 grown on rutin. It is a glycoprotein (46-54% carbohydrate) with N-linked oligo-mannose type glycan chains. The enzyme was resolved in SDS polyacrylamide gels in a diffuse protein band that corresponded to a molecular mass of 130-170 kDa. When purified flavonol 2,4-dioxygenase was heated, it dissociated into three peptides with apparent molecular masses of 63-67 kDa (L), 53-57 kDa (M), and 31-35 kDa (S), which occurred in a molar ratio of 1:1:1, suggesting a LMS structure. Crosslinking led to a 90-97 kDa species, concomitant with the decrease of staining intensity of the 63-67 kDa (L) and the 31-35 kDa (S) peptides. Analysis by matrix-assisted laser desorption/ionization-time of flight-MS showed peaks at m/z approximately 69 600, m/z approximately 51 700, and m/z approximately 26 500 which are presumed to represent the three peptides of flavonol 2,4-dioxygenase, and a broad peak at m/z approximately 96 300, which might correspond to the LS heterodimer as formed in the crosslinking reaction. Based on the estimated molecular mass of 148 kDa, 1 mol of enzyme contained 1.0-1.6 mol of copper. Ethylxanthate, which specifically reduces CuII to CuI ethylxanthate, is a potent inhibitor of flavonol 2,4 dioxygenase. Metal chelating agents (such as diethyldithiocarbamate, diphenylthiocarbazone) strongly inhibited the enzymatic activity, but inactivation was not accompanied by loss of copper. The EPR spectrum of flavonol 2,4-dioxygenase (as isolated) showed the characteristic parameters of a nonblue type 2 CuII protein. The Cu2+ is assumed to interact with four nitrogen ligands, and the CuII complex has a (distorted) square planar geometry. PMID- 10469154 TI - High prevalence of 2-mono- and 2,6-di-substituted manol-terminating sequences among O-glycans released from brain glycopeptides by reductive alkaline hydrolysis. AB - Di- to heptasaccharides isolated from total nondialyzable brain glycopeptides after release by alkaline borohydride treatment have been subjected to mass spectrometric and nuclear magnetic resonance spectroscopic analyses supplemented by TLC-MS analyses of derived neoglycolipids. A family of Manol-terminating oligosaccharides has been revealed which includes novel sequences with a 2, 6 disubstituted Manol: In contrast to the Manol-terminating HNK-1 antigen-positive chains described previously that occur as a minor population [Yuen, C.-T., Chai, W., Loveless, R.W., Lawson, A.M., Margolis, R.U. & Feizi, T. (1997) J. Biol. Chem. 272, 8924-8931], the above oligosaccharides are abundant. The ratio of these compounds to the classical N-acetylgalactosaminitol-terminating oligosaccharides is about 1 : 3. Thus, there appears to be in higher eukaryotes a major alternative pathway related to the yeast-type protein O-mannosylation, the enzymatic basis and functional importance of which now require investigation. PMID- 10469155 TI - Hydrolase and transferase activities of the beta-1,3-exoglucanase of Candida albicans. AB - The exo-beta-1,3-glucanase of Candida albicans (Exg) has a marked specificity for beta-1,3-glucosidic linkages as judged by the kinetic constants for p-nitophenyl beta-glucoside, beta-linked disaccharides of glucose (laminaribiose, gentiobiose, and cellobiose), oligosaccharides of the laminari series, laminarin and pustulan. The kcat/Km ratios for a series of laminari oligosaccharides from -biose to heptaose showed that Exg has an extended substrate-binding site which contains at least five binding sites for sugar residues. Binding at position +2 (the third sugar residue) increases the kcat twofold while positions +3 and +4 lower the Km value further and thereby increase the catalytic efficiency. Exg catalyses an efficient transglucosylation reaction with high concentrations of laminari oligosaccharides which specifically form beta-1,3 linkages and with yields up to 50%. The rate of the transglucosylation is concentration-dependent and can be more than 10 times faster than the hydrolytic reaction with excess donor substrates such as laminaritriose and laminarihexaose. The kinetics of Exg and the predicted substrate-binding site for up to five sugar residues are consistent with a recent structural analysis of the enzyme-binding site. PMID- 10469156 TI - A prokaryotic-type cytidine deaminase from Arabidopsis thaliana gene expression and functional characterization. AB - The gene and cDNA of an Arabidopsis thaliana cytidine deaminase (CDA) were cloned and sequenced. The gene, At-cda1, is located on chromosome 2 and is expressed in all plant tissues tested, although with quantitative differences. Expression analysis suggest that At-cda1 probably codes for the housekeeping cytidine deaminase of Arabidopsis. The gene was functionally expressed in Escherichia coli and the protein, At-CDA1, shows similar enzymatic and substrate specificities as conventional cytidine deaminases: it deaminates cytidine and deoxycytidine and is competitively inhibited by cytosine-containing compounds. Because the protein shows no affinity to RNA, it is not likely to be involved in RNA-editing by C-to U deamination. When compared to cytidine deaminases from other organisms, it becomes clear that At-CDA1 is related, both in sequence and structure, to the CDA of E. coli and other gram-negative bacteria. The eubacterial nature of the Arabidopsis CDA suggests that it is an additional example of a plant gene of endosymbiotic origin. PMID- 10469157 TI - Fibrinolytic properties of activated FXII. AB - Activated factor XII (FXIIa), the initiator of the contact activation system, has been shown to activate plasminogen in a purified system. However, the quantitative role of FXIIa as a plasminogen activator in contact activation dependent fibrinolysis in plasma is still unclear. In this study, the plasminogen activator (PA) activity of FXIIa was examined both in a purified system and in a dextran sulfate euglobulin fraction of plasma by measuring fibrinolysis in a fibrin microtiter plate assay. FXIIa was found to have low PA activity in a purified system. Dextran sulfate potentiated the PA activity of FXIIa about sixfold, but had no effect on the PA activity of smaller fragments of FXIIa, missing the binding domain for negatively charged surfaces. The addition of small amounts of factor XII (FXII) to FXII-deficient plasma induced a large increase in contact activation-dependent PA activity, as measured in a dextran sulfate euglobulin fraction, which may be ascribed to FXII-dependent activation of plasminogen activators like prekallikrein. When more FXII was added, PA activity continued to increase but to a lesser extent. In normal plasma, the addition of FXII also resulted in an increase of contact activation-dependent PA activity. These findings suggested a significant contribution of FXIIa as a direct plasminogen activator. Indeed, at least 20% of contact activation-dependent PA activity could be extracted from a dextran sulfate euglobulin fraction prepared from normal plasma by immunodepletion of FXIIa and therefore be ascribed to direct PA activity of FXIIa. PA activity of endogenous FXIIa immunoadsorped from plasma could only be detected in the presence of dextran sulfate. From these results it is concluded that FXIIa can contribute significantly to fibrinolysis as a plasminogen activator in the presence of a potentiating surface. PMID- 10469158 TI - Transferrins, the mechanism of iron release by ovotransferrin. AB - Iron release from ovotransferrin in acidic media (3 < pH < 6) occurs in at least six kinetic steps. The first is a very fast (G (-3826) polymorphism in the promoter of the gene for uncoupling protein-1 with regard to metabolic parameters and adipocyte transcription factor binding in a large population-based Caucasian cohort. AB - BACKGROUND: The recently described A-->G (-3826) point mutation within the distal region of the UCP-1 promoter is possibly involved in the development of obesity, diabetes and related metabolic disorders. It was the aim of this study to examine the allelic frequency and the prevalence of the three UCP-1 genotypes in a broad caucasian cohort and to investigate the significance of this polymorphism for obesity and diabetes. METHODS: 1020 subjects were randomly chosen from 6450 participants in the Diabetomobile Study. The UCP-1 genotype was determined by genomic PCR and Bcl-I-RFLP analysis in 1020 subjects and tested for association with a variety of metabolic parameters. In addition, the influence of this mutation on adipocyte nuclear factor binding was investigated by electrophoretic mobility shift assays (EMSA). RESULTS: The genotype frequencies in 1020 subjects were: AA genotype, 57.0%; AG genotype, 35.4%; GG genotype, 7.6%; with allelic frequencies of 0.75 for allele A and 0.25 for allele G. No significant differences between the genotypes and age, gender, BMI, leptin, glucose, fasting insulin, C-peptide, HbA1c, diabetes manifestation, total cholesterol, and HDL cholesterol were found. Analysis of the Trp64Arg polymorphism of the beta3 adrenergic receptor in a subgroup of 343 subjects revealed no additive effect to the UCP-1 polymorphism. An yet unknown adipocyte-specific factor of nuclear extracts from 3T3-L1 adipocytes during differentiation is able to bind specifically to the distal UCP-1 promoter region and this binding ability can not be abolished by the mutation. CONCLUSIONS: We determined the genotype and allelic frequency of the UCP-1 promoter polymorphism in the largest known population based study. The results from genotyping demonstrate clearly that this polymorphism does not play a major role in the pathogenesis obesity and diabetes. A yet unknown adipocyte derived and differentiation-dependent regulated transcription factor is able to bind to the distal UCP-1 promoter surrounding 3826 bp. This binding is not affected by presence of the mutation. PMID- 10469166 TI - Serum hyaluronan as a predictor of hepatic regeneration after hepatectomy in humans. AB - BACKGROUND: The capacity for hepatic regeneration after hepatectomy is important for allowing surgeons to determine the appropriate extent of resection. However, conventional preoperative liver function tests are unsatisfactory for estimating the post-operative regenerative capacity of the remnant liver. The aim of this study was to evaluate the relationship between preoperative serum hyaluronan and hepatic regeneration. METHODS: Preoperative serum hyaluronan levels and the hepatic regeneration rate were estimated in 49 patients using computerized tomographic volumetry. The hepatic fibrotic rate was calculated with non-tumorous tissues stained with Azan-Mallory. Immunolocalization of factor VIII-related antigen (FVIIIAg) was examined as a marker for hepatic sinusoidal capillarization. RESULTS: The serum hyaluronan level was significantly correlated with the hepatic regeneration rate (P < 0. 001). Patients with serum hyaluronan levels below 200 ng mL-1 exhibited a significant correlation between the hepatic regeneration rate and the hepatic fibrotic rate. However, patients with serum hyaluronan levels above 200 ng mL-1 did not demonstrate a distinct correlation. The hepatic regeneration rate of patients with FVIIIAg in the liver and serum hyaluronan levels above 200 ng mL-1 were very low compared with those without FVIIIAg (P < 0.001). Multiple regression analysis revealed that serum hyaluronan was a significant predictor of post-operative hepatic regeneration among several clinical variables (r = 0.857, R2 = 0.735). CONCLUSION: It has been suggested that hepatic regeneration is closely related to both hepatic fibrosis and hepatic sinusoidal capillarization. The serum hyaluronan level is regarded as a useful predictor for hepatic regeneration after hepatectomy. PMID- 10469167 TI - Potent synergistic effect of sho-saiko-to, a herbal medicine, during vaccine therapy in a murine model of hepatitis B virus carrier. AB - BACKGROUND: Traditional herbal medicine, sho-saiko-to (TJ-9), improves subjective symptoms, and a recently developed vaccine therapy reduces the viral replication in some chronic hepatitis B virus (HBV)-carriers. The study presented here considers the impact of a combination of vaccine therapy and TJ-9 and the mechanism underlying the therapeutic effect of TJ-9. MATERIALS AND METHODS: HBV transgenic mice (HBV-Tg) expressing similar levels of HBV-related antigens and HBV DNA were used as an animal model of HBV-carrier state, and were assigned to receive either a TJ-9-enriched diet or a monthly injection of vaccine containing hepatitis B surface antigen (HBsAg), or both, for 12 consecutive months. RESULTS: Twelve months after starting the therapy, 9% (1 of 11), 61% (11 of 18), and 100% (10 of 10) of HBV-Tg receiving only the TJ-9-treatment, only the monthly vaccine, and both the TJ-9 and vaccine, respectively, responded to therapy and became completely negative for HBsAg. Spleen lymphocytes and antigen presenting cells (APC) from TJ-9-treated HBV-Tg produced significantly higher levels of IgM, IgG and antibodies to keyhole limpet hemocyanin (KLH) and showed significantly higher stimulatory capacity in allogenic mixed leukocyte reaction (MLR) compared with the spleen cells and APC from HBV-Tg receiving normal diet without TJ-9 (P < 0.05). CONCLUSION: These data confirm the therapeutic role of TJ-9 during HBV infection and inspire optimism of a widespread use of TJ-9 during immune therapies. PMID- 10469168 TI - Angiogenesis extent and expression of matrix metalloproteinase-2 and -9 correlate with upgrading and myometrial invasion in endometrial carcinoma. AB - BACKGROUND: Changes in angiogenesis and expression of extracellular matrix degrading enzymes have been substantiated during tumour changeover and progression. METHODS: Tissues from 64 biopsies of endometrial carcinoma (EC) and 15 biopsies of normal (control) endometrium were investigated immunohistochemically to determine their microvessel number, and by in situ hybridisation to determine the expression of mRNA of the matrix metalloproteinase 2 (MMP-2, gelatinase A) and metalloproteinase-9 (MMP-9 gelatinase B). EC were grouped according to both histological grade (G) G1 to G3 and depth of myometrial (M) invasion M1 to M3. RESULTS: EC as a whole gave significantly higher counts over control endometria. Counts of the G1 group overlapped those of the control group, increased significantly in the G2 and even more in the G3 group. G3 biopsies in particular also displayed most microvessels widely scattered in the tumour tissue, in close association with tumour cells, and as winding and arborized tubes, often dilated in microaneurysmatic segments. Counts also increased in M2 and M3. Expression of the MMP-2 and MMP-9 mRNA, evaluated as percentages of positive biopsies and intensity of expression, were upregulated with the transition from control and G1 groups to G2 and G3, and in relation to advancing depth of invasion. In EC, MMP-2 and MMP-9 mRNA were also expressed by host stromal cells, including microvascular endothelial cells, fibroblasts and macrophages. In the control biopsies, poor expression of MMP-2 mRNA in few endothelial cells and no expression of MMP-9 mRNA were detected. CONCLUSION: These in situ data suggest that angiogenesis and degradation of extracellular matrix occur simultaneously with EC upgrading and advancing depth of invasion, and that EC cells and some host stromal cell populations cooperate in tumour progression. PMID- 10469169 TI - New strategies in anti-vascular cancer therapy. AB - Angiogenesis is a critical step in the progression of tumours from dormancy to a clinical relevant cancer. Inhibition of this process is one of the most promising new anti-cancer strategies. To develop new drugs that interfere with the cascade of events required for the formation of new blood vessels, insight into this process is essential. Here, we discuss the molecular basis of angiogenesis and the concepts of vascular targeting. Furthermore new strategies will be discussed to discover surface markers on endothelial cells that confer sufficient specificity for targeted intervention in the tumour vasculature. PMID- 10469170 TI - The role of HSP90 in evolution. AB - A mechanism by which morphological mutations are stored without expressing phenotypes was unraveled by Rutherford & Lindquist (1998) through genetic studies of Hsp83 (HSP90) in Drosophila. Cryptic mutations are essentially neutral and therefore evolve in the absence of selective constraint. A shift from neutral mutations to selective mutations is induced when flies are exposed to environmental stress. This is a step toward understanding macroevolution in molecular terms. PMID- 10469171 TI - Competitive interaction of seven in absentia homolog-1A and Ca2+/calmodulin with the cytoplasmic tail of group 1 metabotropic glutamate receptors. AB - BACKGROUND: Group 1 metabotropic glutamate receptors (mGluR1 and mGluR5) are coupled to inositol trisphosphate/Ca2+ signaling via G proteins and play an important role in excitatory synaptic transmission. To explore the regulation of group 1 mGluR function, we applied the yeast two-hybrid system using the intracellular carboxy-terminal domain of group 1 mGluRs (group 1 ct-mGluRs) and attempted to identify novel protein-protein interactions of group 1 mGluRs. RESULTS: The two-hybrid screening revealed a specific interaction between group 1 ct-mGluRs and Siah-1A, the mammalian homolog of Drosophila seven in absentia which is involved in photoreceptor cell differentiation via the ubiquitin/proteasome-dependent mechanism. This interaction occurs within a homologous 27-28 amino acid stretch within group 1 ct-mGluRs and requires the latter two-thirds of Siah-1A. Following coexpression in COS-7 cells, myc-tagged Siah-1A was coimmunoprecipitated with the flag-tagged ct-mGluR1 by anti-flag antibody. Furthermore, in vitro binding revealed that Siah-1A and Ca2+/calmodulin (CaM) binding sites overlap, such that Siah-1A binding is competitively inhibited by CaM in a Ca2+-dependent manner. CONCLUSIONS: The results demonstrate a direct interaction between group 1 mGluRs and Siah-1A and suggest a novel modulatory mechanism mediated by a competitive interaction between Ca2+/CaM and Siah-1A. PMID- 10469172 TI - Negative regulation of the pts operon by Mlc: mechanism underlying glucose induction in Escherichia coli. AB - BACKGROUND: The pts operon of Escherichia coli consists of three genes ptsH, ptsI and crr, each encoding for central components of the phosphoenolpyruvate: carbohydrate phosphotransferase system, HPr, enzyme I and IIAGlc, respectively. Transcription of the pts operon is stimulated when glucose is present in the culture medium. One of the two major promoters, P0, is responsible for this glucose induction. However, no regulatory protein responsible for the glucose induction of the pts operon has been identified yet and molecular mechanism by which glucose stimulates the pts transcription is not known. RESULTS: We found by Northern blotting that the pts mRNA levels in cells lacking Mlc, a new global repressor of carbohydrate metabolism, were increased without external glucose and that the addition of glucose had no effect on the pts mRNA levels in the mutant cells. Western blotting revealed that the enzyme I level in the mlc- cells was also elevated without glucose and no further increase in the enzyme I level was observed in the presence of glucose. S1 analysis revealed that transcription of the glucose-sensitive promoter, P0, occurs constitutively in the mlc- cells independently from the external glucose. In vitro transcription studies indicated that Mlc strongly inhibited P0 transcription. DNase I footprinting experiment revealed that Mlc bound to P0 promoter region to prevent RNA polymerase binding at P0. CONCLUSION: We conclude that Mlc is a repressor for the pts transcription acting as a major regulatory protein involved in the glucose induction of pts operon. We propose that glucose induces the pts transcription by modulating the Mlc activity. The mechanism by which glucose modulates the Mlc action remains to be studied. PMID- 10469174 TI - Identification of the core domain and the secondary structure of the transcriptional coactivator MBF1. AB - BACKGROUND: Multiprotein bridging factor 1 (MBF1) is a transcriptional coactivator necessary for transcriptional activation caused by DNA binding activators, such as FTZ-F1 and GCN4. MBF1 bridges the DNA-binding regions of these activators and the TATA-box binding protein (TBP), suggesting that MBF1 functions by recruiting TBP to promoters where the activators are bound. In addition, MBF1 stimulates DNA binding activities of the activators to their recognition sites. To date, little is known about structures of coactivators that bind to TBP. RESULTS: The two-dimensional (2D) 1H-15N correlation spectrum of 15N labeled MBF1 indicated that MBF1 consists of both flexible and well structured parts. Limited digestion of MBF1 by alpha-chymotrypsin yielded a approximately 9 kDa fragment. N-terminal sequence analysis and NMR measurements revealed that this fragment originates from the C-terminal 80 residues of MBF1 and form a well structured C-terminal domain of MBF1, MBF1CTD. As previous deletion analyses have shown that MBF1CTD is capable of binding to TBP, it is suggested that MBF1CTD is the TBP binding domain of MBF1. Sequential assignments have been obtained by means of three-dimensional (3D) and four dimensional (4D) heteronuclear correlation spectroscopies, and then the secondary structure of MBF1CTD was determined. As a result, MBF1CTD was shown to contain four amphipathic helices and a conserved C-terminal region. Asp106 which is assumed to be responsible for the binding to TBP is located at the hydrophilic side of the third helix. CONCLUSIONS: Structural analyses revealed that MBF1 consists of two structurally different domains. A N-terminal region is indispensable for the binding to activators, and does not form a well defined structure. In contrast, the C terminal 80 residues, which is capable of binding to TBP by itself, form a well structured domain, MBF1CTD. MBF1CTD is made up of four amphipathic helices and a conserved C-terminal tail. A putative TBP binding residue is located on the hydrophilic surface of the third helix. PMID- 10469173 TI - Changes in nuclear morphology during apoptosis correlate with vimentin cleavage by different caspases located either upstream or downstream of Bcl-2 action. AB - BACKGROUND: Upon Fas stimulation, procaspase-8 is recruited to the death-inducing signalling complex where autoactivation of caspase-8 occurs. Active caspase-8 can directly activate downstream caspases (e.g. caspase-3, 6, and 7) for the execution of apoptosis (mitochondria-independent pathway), while caspase-8 can also lead to executioner caspase activation through mitochondrial damage (mitochondria-dependent pathway). Caspase activation results in the dismantling of intracellular structure through specific proteolysis. RESULTS: We have found that an intermediate filament protein, vimentin, is cleaved at multiple sites by caspases at an early stage of apoptosis in Jurkat cells. The sequences of the two major cleavage sites in vimentin (IDVD/V and DSVD/F) suggested that these sites are cleaved by caspase-8 and caspase-3, respectively, or by close homologues of these proteases. The IDVD/V site can be cleaved by caspase-8 in vitro, and its cleavage is less sensitive to DEVD-CHO and Bcl-2 over-expression than that of the DSVD/F site in Jurkat cells. Over-expression of a mutant vimentin which was insensitive to caspase cleavage at these sites delayed the appearance of apoptotic nuclei in Jurkat cells. CONCLUSION: The specific cleavage of vimentin can be used as an apoptotic marker of both apical- and mitochondria-dependent caspase activation. Apoptotic cleavage of vimentin most likely results in disruption of its filamentous structure, which may facilitate nuclear condensation and subsequent fragmentation through disruption of the cytoskeletal network. PMID- 10469175 TI - Desmopressin in the treatment of women's bleeding disorders. AB - Bleeding disorders in women are an underestimated problem that deserves increased attention. About 9%-14% of females have menorrhagia and, amongst them, there is a significant over-representation of von Willebrand disease (VWD), with a prevalence of 13% in this group as compared with about 1% in the general population. The bleeding disorder has not been diagnosed in most of these women and they may therefore be withheld from treatment with desmopressin, which is effective in most cases of VWD and also in platelet dysfunctions and mild factor VIII deficiency. This paper is a review of the haemostatic use of desmopressin with special reference to women's bleeding disorders, the mechanisms of action, modes of administration, clinical indications, dosage recommendations, and hospital or home treatment. Desmopressin stimulates endogenous release of FVIII and von Willebrand factor (VWF), it increases platelet adhesiveness and shortens bleeding time. It can be given as intravenous or subcutaneous injection, but the intranasal spray is probably the most practical mode of administration for females with bleeding disorders as it is simple to administer and suitable for home treatment. The spray has been used successfully in connection with menorrhagia and other bleeding symptoms. PMID- 10469176 TI - Haemophilia B carrier detection by factor IX:C analysis; no impact of the type of mutation or severity of disorder. AB - Haemophilia B, an X-linked recessive bleeding disorder characterized by lack or deficiency of factor IX, has been shown to be caused by any of a variety of DNA abnormalities (partial or total deletions, nonsense or missense mutations). Since in most countries carrier detection is based on factor IX coagulant activity (FIX:C) assay, this study was designed to determine whether carriers' FIX:C values are dependent on the severity of haemophilia (mild, moderate or severe) or on the genetic anomaly in the family. FIX:C concentrations were studied in 28 obligate carriers, 39 women known to carry the mutation and 33 verified noncarriers subgrouped by severity of disorder or genetic anomaly. No significant subgroup differences in FIX:C values were found, thus suggesting the level of FIX:C concentrations in carriers to be unaffected by the severity of haemophilia, or by its expression (i.e. deficient or dysfunctional factor IX). The specificity and sensitivity of FIX:C analysis for the purpose of carrier diagnosis was judged by receiver operating characteristic curve analysis, where an FIX:C cut-off level of 75 IU dL-1 was found to be optimal (sensitivity 93% and specificity 88%). PMID- 10469177 TI - Carrier detection in haemophilia A families: comparison of conventional coagulation parameters with DNA polymorphism analysis - first report from India. AB - Linear discriminants that include data on factor VIII:C and von Willebrand factor antigen levels are well-established tools in estimating the probability of carriership in haemophilia A families. A comparison between the conventional coagulation data, i.e. the ratio of factor VIII:C and von Willebrand factor antigen, and the DNA analysis techniques was made in 98 confirmed carriers (39 obligatory, 69 detected by gene tracking analysis) and 71 normal age matched females who did not have any history of bleeding and were not taking any drug. The lowest misclassification rate, i.e. 7% among the carriers, was seen when a cut-off value of 0.7 was chosen. In the case of normals, all were outside this cut-off value. Thus, it was considered as a workable reference value for classifying the carriers in haemophilia A families in our laboratory. We conclude that the optimal service for haemophilia A carrier diagnosis must include above coagulation test probabilities as well as DNA marker studies. However, it is recommended that the smaller laboratories in developing countries can benefit immensely by only establishing factor VIII:C and von Willebrand factor antigen estimation. PMID- 10469178 TI - The impact of inhibitors on the cost of clotting factor replacement therapy in Haemophilia A in Canada. AB - A retrospective case controlled study was performed to determine the comparative costs of clotting factor concentrate therapy for haemophilia A patients with and without inhibitors. We examined treatment records for a 3.5-year period for nine patients with factor VIII inhibitors and nine noninhibitor control patients matched for age and severity of disease. Inhibitor patients used FEIBA, porcine factor VIII, recombinant factor VIIa, and recombinant factor VIII over the study period. Controls used recombinant factor VIII and small amounts of monoclonal antibody purified plasma-derived factor VIII and DDAVP. The total and mean cost for treating the nine inhibitor patients was 2.25-fold greater than the cost for treating the controls. However, in six of the nine pairs the replacement product costs were actually less for the control patient than for the inhibitor patient, and the median cost of concentrates was comparable in the two groups (CDN$150 686 and $133 342 for inhibitor and control patients, respectively). This discrepancy was largely accounted for by a single inhibitor patient who required frequent hospitalizations for severe bleeding episodes. This individual, who did not receive an immune tolerance protocol, accounted for 62% of the total costs for the entire inhibitor group. In summary, over the study period the cost of haemostatic therapy for most inhibitor patients did not exceed that of control patients. However, due to the high per-unit cost of the products used to treat inhibitor patients, the effect of individual outliers greatly magnifies the overall costs of treating patients with this complication. PMID- 10469179 TI - Recombinant factor VIIa for patients with inhibitors to factor VIII or IX or factor VII deficiency. AB - Inhibitors to factor VIII (FVIII) or IX (FIX) in patients with haemophilia A or B create a challenging problem for the treatment of these patients. Recombinant FVIIa (rFVIIa; NovoSeven, Novo Nordisk A/S, Bagsvaerd, Denmark) is a realistic treatment option, owing to its specific mode of action and lack of immunogenicity. This was a multicentre, open-label, compassionate-use trial in patients with severe haemophilia A (FVIII:C < 1%) or B (FIX:C < 1%) with inhibitors, acquired antibodies to FVIII or FIX, or FVII deficiency (FVII:C < 5%), for whom alternative therapies had failed or were contraindicated. Patients received rFVIIa treatment for life- or limb-threatening bleeding episodes or for coverage during essential surgery. The mean rFVIIa dose was approximately 90 microg kg-1 for haemophilia A/B and acquired inhibitor patients, and 25 microg kg 1 for FVII-deficient patients. Efficacy data for 67 treatment episodes (45 bleeding episodes, 22 surgical procedures) are presented; seven patients were treated for a concurrent serious bleeding episode and surgical procedure. At the end of treatment, rFVIIa was effective or partially effective in 85% of serious bleeding episodes. During surgery, bleeding was assessed as none or less than or equivalent to normal in 91% of surgical procedures; postoperatively, 91% of procedures were associated with no or minimal oozing. During 60 separate treatment episodes, 26 adverse events (22 nonserious, four serious) were reported in 15 patients, during 17 bleeding episodes or surgical procedures. Only 10 were considered as having a possible, probable, or unknown relationship with rFVIIa; of these, fever (n=2) and thrombophlebitis (n=3) were the most common. There was no evidence of disseminated intravascular coagulation. In conclusion, rFVIIa is an effective, well-tolerated treatment for serious bleeding episodes and bleeding associated with surgical procedures in patients with severe haemophilia A/B with inhibitors, acquired inhibitors, or FVII deficiency. PMID- 10469180 TI - Pharmacokinetics and acute tolerance of a double virus inactivated plasma derived factor VIII concentrate. AB - To further reduce the risk of possible viral disease transmission, an additional virucidal step was performed in the manufacturing process of a solvent/detergent treated factor VIII concentrate, which consisted of heating the lyophilized preparation at 100 degrees C for 30 min (Emoclot DI; ISI, Italy). Because thermal treatment may modify factor VIII bioavailability, the pharmacokinetic parameters and the acute tolerance of the single viral inactivated concentrate (preparation A) were compared with that of the double viral inactivated one (preparation B). Fifteen patients with severe haemophilia A and positive for HAV Ab were enrolled in a double-blind cross-over study and injected with 32.5 IU kg-1 of preparation A and 27 IU kg-1 of the preparation B. No significant differences between terminal half-life, area under the curve/dose, clearance/kg, volume of distribution at the steady state, in vivo recovery and acute tolerance of the two preparations was observed. The only statistical difference was restricted to Cmax. PMID- 10469181 TI - Bleeding episodes in HIV-positive patients taking HIV protease inhibitors: a case series. AB - In July 1996 the Food and Drug Administration (FDA) alerted healthcare providers about 15 case reports of spontaneous bleeding episodes in HIV-positive haemophiliacs taking HIV protease inhibitors. In order to characterize the bleeding associated with HIV protease inhibitor therapy, the FDA's spontaneous adverse event reporting system was searched through 28 February 1997. The bleeding episode reporting rate for persons with haemophilia was compared for HIV protease inhibitors and zidovudine, and the characteristics of haemorrhagic events were compared between individuals with and without haemophilia. There was a substantial predominance of bleeding episodes for haemophiliacs taking HIV protease inhibitors (39 of 67; 58%) as compared with zidovudine (two of 63; 3.2%). A comparison of 39 reports of bleeding in haemophiliacs with 28 in non haemophiliacs revealed similarities in time to event and type of HIV protease inhibitor implicated, but differences were present concerning location of bleeding and outcome. A greater proportion of haemophiliacs had resolution of their bleeding following discontinuation of their HIV protease inhibitor and recurrence of bleeding following rechallenge, as compared with non-haemophiliacs. HIV-positive haemophiliacs appear to be at an elevated risk of bleeding while taking HIV protease inhibitors, but these medications may predispose all individuals to such complications. PMID- 10469182 TI - Acute renal failure in acquired haemophilia following the use of high dose intravenous immunoglobulin. AB - High dose intravenous immunoglobulin (IVIG) is a useful form of treatment in patients with acquired haemophilia, a rare bleeding disorder due to auto antibodies to factor VIII. We describe a patient with acquired haemophilia who developed acute renal failure rapidly after treatment with a high sucrose content IVIG preparation. We speculate that the sucrose content of the preparation was responsible for the renal failure and make recommendations on the use of IVIG in patients with acquired haemophilia. PMID- 10469183 TI - Use of recombinant factor VIIa (NovoSeven) in a haemophilia A patient with inhibitor in Kuwait. AB - Development of inhibitors is a known complication in some haemophiliacs receiving coagulation factor replacement therapy. We report on the successful management of a young boy with haemophilia A with inhibitor using recombinant factor VIIa. We had failed to control bleeding in this patient following his circumcision, despite infusion with high doses of factor VIII concentrate for 2 weeks. Recombinant factor VIIa is a useful 'factor VIII bypassing agent' for the control of bleeding in patients with haemophilia A and B who develop inhibitors. We suggest that severely affected haemophiliacs should be absolved of ritual circumcision as a protective measure against what might become a life-threatening haemorrhage - especially in those with inhibitors. PMID- 10469184 TI - Orthotopic liver transplantation in a patient with severe haemophilia A and with advanced liver cirrhosis. AB - A patient with severe haemophilia A underwent orthotopic liver transplantation because of changes correlated to end-stage liver cirrhosis due to hepatitis B, C and D infection. Replacement therapy was carried out for 4 days and the clinical course was uneventful. At the time of reporting the patient has a normal working life. FVIII plasma concentration is normal. The indirect hyperbilirubinaemia may be related to the Gilbert's anomaly of the donor. PMID- 10469185 TI - Safety and efficacy of three arthroscopic procedures using Holmium: Yag laser in two high-responder haemophiliacs. AB - We report here on the efficacy and safety of three arthroscopic procedures using a Holmium: Yag laser in two high-responder haemophiliacs. The two patients were treated with an activated prothrombin complex concentrate (FEIBA; Immuno, Vienna, Austria). Treatment was started just before surgery and continued for 4-8 days. On one occasion antifibrinolytics were concomitantly used without thromboembolic complication. Post-operative blood loss was slight, joint mobility was rapidly acceptable and full weight bearing without pain was possible on day 4. Such a procedure would appear to be superior to conventional arthroscopic synovectomy utilizing mechanical devices in haemophiliacs, because it might improve the quality of local haemostasis and the rapidity of post-operative recovery. In addition, it is also the technical procedure of choice in haemophilic patients with inhibitors who need synovectomy. PMID- 10469186 TI - Inter-laboratory variability of the measurement of low titer factor VIII:C inhibitor in haemophiliacs: improvement by the Nijmegen modification of the Bethesda assay and the use of common lyophilized plasmas. PMID- 10469187 TI - CagA and VacA: virulence factors of Helicobacter pylori in Thai patients with gastroduodenal diseases. AB - BACKGROUND: The aim of this study was to assess the seroprevalence of cytotoxin associated gene A (cagA) and vacuolating cytotoxin gene A (vacA) of Helicobacter pylori in selected Thai populations with specific gastroduodenal diseases. MATERIALS AND METHODS: The immunoblot assay was used to detect serum antibodies against CagA and VacA obtained from the following patients: 87 cases of nonulcer dyspepsia (NUD), 61 cases of duodenal ulcer (DU), 49 cases of gastric ulcer (GU), and 10 cases of gastric cancer (GC). RESULTS: Serum antibodies to CagA were detected in 75.4% of all patients (70. 1% of NUD, 78.7% of DU, 77.6% of GU, and 90% of GC). Although the prevalence of CagA seropositivity in GC patients was higher than in the other three groups, the difference was not statistically significant (p >.05). CONCLUSIONS: The high seroprevalence of the CagA-positive H. pylori strain in patients with peptic ulcer, GC, and NUD indicates that this strain is common in Thai patients with gastroduodenal diseases. Furthermore, phenotypic classification of H. pylori into type 1 (CagA-positive, VacA-positive) and type 2 (CagA-negative, VacA-negative) is not a useful marker for screening patients with severe forms of gastroduodenal diseases. PMID- 10469188 TI - Antigenic mimicry between Helicobacter pylori and gastric mucosa: failure to implicate heat-shock protein Hsp60 using immunoelectron microscopy. AB - BACKGROUND: Helicobacter pylori infection induces autoantibodies that cross-react with human gastric mucosa from infected individuals. Candidates for the antigens responsible for molecular mimicry causing autoreactivity include the heat-shock protein HspB (Hsp60, sometimes called Hsp54) or Lewis x and Lewis y carbohydrate antigens. OBJECTIVE: Our goal was to investigate the involvement of HspB (Hsp60) in autoreactivity between H. pylori and gastric biopsy tissue. MATERIALS AND METHODS: Immunoelectron microscopy was used to study cross-reactivity among biopsy tissues from a patient with gastritis, gastric ulcer, and duodenal ulcer and his own serum as well as reactivity with serum raised against HspB from H. pylori and monoclonal antibodies against Lewis antigens. RESULTS: The patient serum reacted with gastric mucosa, and the antibodies involved were predominantly IgG. Antibody raised to H. pylori HspB (Hsp60) reacted only with H. pylori cells but not with gastric mucosal tissue. In contrast, monoclonal antibodies specific for Lewis x and Lewis y antigens reacted with both H. pylori and human gastric epithelial tissue. CONCLUSIONS: Hsp60 (Hsp54) is unlikely to be involved in autoreactivity seen in individuals infected with H. pylori. In contrast, we could not rule out the role of Lewis x and Lewis y carbohydrate antigens, expressed as a component of H. pylori lipopolysaccharides, in molecular mimicry and autoantibody production. PMID- 10469189 TI - Different penicillin-binding protein profiles in amoxicillin-resistant Helicobacter pylori. AB - BACKGROUND: The beta-lactam group of antibiotics kills bacteria by inhibiting the terminal stages of peptidoglycan metabolism. We have recently identified amoxicillin-resistant Helicobacter pylori, none of which expressed beta lactamase. Penicillin-binding proteins (PBPs) represent a group of target enzymes for the beta-lactam antibiotic family, and alterations in PBPs have been described in other penicillin-resistant bacteria. The amoxicillin-resistant phenotype characteristically was lost after freezing but could be restored by consecutive transfers into gradient plates. MATERIALS AND METHODS: To determine whether amoxicillin resistance in H. pylori was related to alterations in any of the H. pylori PBPs, five H. pylori strains resistant to amoxicillin and three amoxicillin-sensitive strains were tested. PBPs were extracted from bacteria grown to logarithmic phase, labeled in vivo with 3H-benzylpenicillin, and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. Four main PBPs were separated from all amoxicillin-sensitive H. pylori strains. RESULTS: Only three of the four main PBPs were found in the amoxicillin-resistant H. pylori strains. The differentially detectable PBP (PBP D) had an apparent molecular weight of 30 to 32 kD. CONCLUSION: These results suggest that PBP D might play a role in the amoxicillin-resistant phenotype of H. pylori strains lacking beta-lactamase activity. PMID- 10469190 TI - Role of Hpn and NixA of Helicobacter pylori in susceptibility and resistance to bismuth and other metal ions. AB - BACKGROUND: Helicobacter pylori produces Hpn, a 60-amino acid, histidine-rich protein that avidly binds nickel and zinc ions, and NixA, a high-affinity nickel transporter in the cytoplasmic membrane. We tested the hypothesis that Hpn and NixA govern susceptibility to metal ions in H. pylori. MATERIALS AND METHODS: Hpn negative mutants of four H. pylori strains were constructed by standard allelic exchange techniques to yield isogenic Hpn+/Hpn-deficient pairs. A metal concentration that inhibited growth by 50% (IC50) was calculated for Ni2+, Zn2+, Cu2+, and Co2+ by comparing OD600 of cultures in metal-supplemented and control media. RESULTS: Among all four pairs of isogenic strains, the tolerance for Ni2+ was reduced significantly (p <.001) in the Hpn mutants; the mean IC50 value for wild-type strains was 1.9 mM; for the mutant, it was 0.8 mM. In contrast, growth inhibition by Zn2+ was identical within the fours pairs, as was Cu2+ and Co2+ tolerance in one pair tested. We also found that deletion of the hpn gene increases susceptibility to therapeutic forms of bismuth by testing a mutant and wild-type pair with ranitidine bismuth citrate, bismuth citrate, and four antibiotics. Minimal inhibitory concentrations of ranitidine bismuth citrate dropped from 9.2 to 2.3 microg/ml, and those of bismuth citrate dropped from 7.4 to 3.2 microg/ml (p <.05 for both comparisons), while susceptibility to the antibiotics was unaffected. Disruption of the nixA gene encoding the specific Ni2+ transport protein of H. pylori did not change susceptibility to bismuth. CONCLUSION: We concluded that bacteria lacking Hpn, cultured in vitro, are more susceptible than is the wild type to bismuth and Ni2+. PMID- 10469191 TI - Comparison of meal-stimulated serum gastrin response in Helicobacter pylori positive duodenal ulcer and asymptomatic volunteers with and without H. pylori infection. AB - BACKGROUND: Duodenal ulcer (DU) patients exhibit raised postprandial gastrin release as compared to that in healthy controls. It is believed that serum pepsinogen I (PG I) concentration reflects the chief cell mass and that hyperpepsinogenemia I plays an important role in the pathogenesis of DU. Currently, strong evidence suggests that Helicobacter pylori (H. pylori) infection plays an important role in the pathogenesis of DU. MATERIALS AND METHODS: Subjects consisted of 15 patients with H. pylori-positive DU, 10 H. pylori-positive volunteers, and 35 H. pylori-negative volunteers. Blood samples were taken before and at 15, 30, and 60 minutes after eating the test meal, which consisted of 100 gm rice, 130 gm chicken, and 1 egg. The 1-hour integrated gastrin response (IGR) was taken as the area under the serum gastrin time curve, calculated by the trapezoid method. Serum gastrin (SG) and fasting serum PG I concentrations were measured by radioimmunoassay. RESULTS: Meal-stimulated SG response and fasting PG I concentration were significantly higher in DU patients than in H. pylori-positive and -negative volunteers. The DU patients were divided into two groups in accordance with their IGR levels as follows: hyper-IGR and normo-IGR. Serum PG I concentration was significantly higher in the hyper-IGR than in the normo-IGR group. CONCLUSIONS: The DU patients differed in some way (other than H. pylori infection) from the H. pylori-positive healthy volunteers. The fact that hyper-IGR DU patients had higher serum PG I concentrations suggests that patients in this group may be acid hypersecretors. PMID- 10469192 TI - Comparative treatment of Helicobacter pylori-positive duodenal ulcer using pantoprazole at low and high doses versus omeprazole in triple therapy. AB - BACKGROUND: Helicobacter pylori eradication has become the standard treatment for peptic ulcer disease. H. pylori-eradicating triple therapy with omeprazole plus two antibiotics has been used until recently; however, the efficacy of pantoprazole and antibiotics for H. pylori eradication has not been researched thoroughly until now. The aim of this randomized clinical trial was to verify the efficacy of triple oral therapy comparing the effects of pantoprazole using two different doses versus omeprazole twice daily in H. pylori eradication, in ulcer healing and relapses, and in gastritis improvement. MATERIALS AND METHODS: We enrolled 243 patients with H. pylori-positive duodenal ulcer and randomized them into three treatment groups: 84 patients (group Ome40) were assigned to receive omeprazole, 20 mg twice daily, plus amoxicillin, 1 gm twice daily, and clarithromycin, 500 mg twice daily for 10 days; 79 patients (group Pan40) were treated with pantoprazole, 40 mg daily, plus amoxicillin and clarithromycin at the same doses as those of group Ome40; and 80 patients (group Pan80) were treated with pantoprazole, 40 mg twice daily, plus amoxicillin and clarithromycin at the same doses as those of group Ome40. RESULTS: Ulcer healing was observed in 81 of 84 patients (96.4%) in group Ome40; in 66 of 79 patients (83.5%) in group Pan40; and in 77 of 80 patients (96.2%) in group Pan80. H. pylori was eradicated in 79 of 84 patients (94%) in group Ome40; in 63 of 79 patients (79.7%) in group Pan40; and in 75 of 80 patients (93.7%) in group Pan80. CONCLUSIONS: We found that 10-day triple therapy with amoxicillin, clarithromycin, and either pantoprazole, 80 mg daily, or omeprazole, 40 mg daily, is highly effective in ulcer healing and is very well tolerated, achieving the 90% cure recommended for an ideal first-line anti-H. pylori positive duodenal ulcer treatment regimen. PMID- 10469193 TI - Host determinants of Helicobacter pylori infection and its clinical outcome. AB - Greater than one-half of the world's population harbors Helicobacter pylori. The majority of infected individuals, however, remain asymptomatic, with only 10% to 20% developing diseases, including peptic ulcer disease, gastric cancer, and gastric mucosa-associated lymphoid tissue lymphoma. This article reviews host factors that may predispose an individual to both the acquisition of H. pylori infection and subsequent clinical outcome. Individuals with specific blood group antigens and human leukocyte antigen genotypes may be more susceptible to H. pylori infection. Additional factors, such as the age of acquisition, the host immune response, the site of infection, acid secretion, and interactions with nonhost factors (including bacterial virulence factors and environmental influences) may play a role in determining clinical outcome. Further investigation is required to clarify the mechanisms by which these interactions occur and, more critically, to determine their relative importance. This knowledge will enable the identification of individuals at risk of developing clinical disease with H. pylori infection. PMID- 10469194 TI - Efficacy of conventional endoscopic disinfection and sterilization methods against Helicobacter pylori contamination. AB - BACKGROUND: Iatrogenic transmission of Helicobacter pylori via contaminated endoscopic devices is well documented. Despite the prevalence of this infectious agent, few controlled studies have investigated the major factors that impact on reprocessing of endoscopes contaminated with H. pylori. MATERIALS AND METHODS: An endoscope (Pentax) was contaminated with 108 cfu/ml of H. pylori in 5% bovine calf serum as standardized inoculum. The endoscope then was passed through one of eight arms (five repetitions per arm = 40 total runs), as follows: 1, recovery control (no cleaning or disinfection); 2, manual cleaning alone; 3-5, manual precleaning followed by either 10-, 20-, or 45-minute exposure to 2% glutaraldehyde and ethanol (ETOH) drying; 6, manual cleaning followed by automated reprocessing by STERIS System; 7 and 8, automated reprocessing by STERIS with and without active peracetic acid sterilant (wash-off control). Suction-biopsy channels and air-water channels were harvested for microbiological culture. RESULTS: Control runs recovered more than 1 x 106 cfu per site, confirming the viability of the test organism and the adequacy of the biological burden for challenge. When instruments underwent manual cleaning alone (without subsequent disinfection), test organisms remained in 40% of runs at the air-water site. Manual cleaning followed by 10-, 20-, or 45-minute glutaraldehyde exposure and ETOH drying removed all test organisms from all sites in all runs (i.e., 100% disinfection). The automated STERIS system with or without active peracetic acid sterilant also removed all test organisms from all sites in all runs, as did manual cleaning followed by STERIS use. CONCLUSION: Manual cleaning alone does not effectively remove H. pylori from an endoscope. Current joint association recommendations for minimal disinfection (manual cleaning followed by at least 20 minutes of immersion in glutaraldehyde and ETOH drying) are effective in preventing cross-transmission of H. pylori. Reprocessing using the automated STERIS system according to manufacturer's recommendations also is highly effective in sterilizing endoscopes contaminated with H. pylori. PMID- 10469195 TI - Effect of smoking and histological gastritis severity on the rate of H. pylori eradication with omeprazole, amoxicillin, and clarithromycin. AB - BACKGROUND: The combination of omeprazole, amoxicillin, and clarithromycin is a common regimen against Helicobacter pylori. Several recent studies have shown that smoking, high intragastric acidity, and the degree of histological gastritis are associated with H. pylori eradication failure. MATERIALS AND METHODS: One hundred and thirty-seven H. pylori-positive patients were treated with a 1-week regimen composed of omeprazole, 20 mg once daily; amoxicillin, 500 mg; and clarithromycin, 200 mg thrice daily. Success of the treatment was evaluated by histology and the 13C-urea breath test at least 4 weeks after completion of therapy. Data about age, gender, alcohol intake, smoking habits, and previous proton pump inhibitor intake were collected in patient interviews. We evaluated fasting gastric pH and the degree of histological gastritis before eradication of H. pylori. RESULTS: The overall eradication of H. pylori at 4 weeks was successful in 98 of 137 patients (72%). On the multivariate analysis, a low grade of inflammation in the antrum (p tgaag. The mutations were confirmed either by restriction endonuclease or sequence-specific oligonucleotide hybridization analysis. In addition, an analysis of 110 alleles from 74 unrelated normal individuals demonstrated that the DNA sequence changes were not common polymorphisms. All of the mutations predict truncated chloride channels that are likely to result in a functional loss. Thus, our findings expand the spectrum of CLCN5 mutations associated with Dent's disease and the results will help to elucidate further the functional domains of this novel chloride channel. PMID- 10469282 TI - 24,25-dihydroxyvitamin D3 suppresses the rapid actions of 1, 25-dihydroxyvitamin D3 and parathyroid hormone on calcium transport in chick intestine. AB - Studies were undertaken to determine whether 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) modulates the rapid effects of 1, 25-dihydroxyvitamin D3 (1,25(OH)2D3) and parathyroid hormone (PTH) on calcium transport in the perfused chick intestine. Perfusion with control media resulted in a transport ratio (treated/average basal) of 1.07 +/- 0.06 at t = 40 minutes, while perfusion with 65, 130, 300, or 650 pM 1,25(OH)2D3 yielded ratios of 1.92 +/- 0.23, 2.6 +/- 0.4, 2.8 +/- 0.08, and 3.34 +/- 0.37, respectively. Simultaneous perfusion with each of these doses and 6.5 nM 24,25(OH)2D3 reduced treated/average basal ratios to approximately 1.4 after 40 minutes of perfusion. Vascular perfusion with 65 pM bovine PTH [bPTH(1-34)] stimulated intestinal calcium transport ratios to 3.0 +/- 0.5 after 40 minutes, while the inclusion of 6.5 nM 24,25(OH)2D3 reduced ratios at this time point to 0.56 +/- 0.19. To investigate the effect of these agents on signal transduction, isolated intestinal cells were monitored for intracellular calcium changes using the indicator dye fura-2. After establishing a stable baseline, addition of 130 pM 1,25(OH)2D3 induced rapid calcium oscillations. Intestinal cells exposed to 6.5 nM 24,25(OH)2D3 also exhibited rapid oscillations in fluorescence, which were not further altered by subsequent addition of 1,25(OH)2D3. Incubation of isolated cells with 130 pM 1,25(OH)2D3 was found to increase protein kinase C (PKC) activity within 5 minutes, and protein kinase A (PKA) activity within 7 minutes. Exposure of cells to 65 pM bPTH(1-34) had minimal effect on PKC activity, but resulted in pronounced increases in PKA activity. Stimulation of protein kinases by either secosteroid or peptide hormone was inhibited in the presence of 6.5 nM 24,25(OH)2D3. It is concluded that 24,25(OH)2D3 may exert endocrine actions on intestine. PMID- 10469283 TI - Expression of thyroid hormone receptor isoforms in rat growth plate cartilage in vivo. AB - Although thyroid hormone has been known for many years to be a potent regulator of skeletal maturation in vivo, it has not definitively been determined whether this effect is a result of a direct or indirect action of the hormone. Previous in vivo studies have suggested that thyroid hormone may stimulate longitudinal bone growth by increasing the secretion of growth hormone; however, growth hormone alone is unable to stimulate cartilage maturation. There are also indications that thyroid hormone is able to act directly on growth plate chondrocytes through growth hormone-independent mechanisms. In this study, we demonstrate that rat growth plate chondrocytes in vivo express genes encoding three of the four isoforms of the thyroid hormone receptors described to date, but the corresponding protein can only be detected for the TRalpha1 and TRbeta1 isoforms of the receptor. As has been noted in other tissues, there is generally poor correlation between the mRNA levels for each isoform and the relative amount of corresponding protein as measured by immunoblotting, suggesting the possibility that receptor expression may be regulated by post-transcriptional mechanisms. PMID- 10469284 TI - A dose-finding study of zoledronate in hypercalcemic cancer patients. AB - Zoledronate is a new heterocyclic imidazole bisphosphonate that is the most potent bisphosphonate administered in humans because it is 100-850 times more potent than pamidronate, according to in vitro or animal models of bone resorption. We conducted an open-label, dose-finding, single-dose phase I study in tumor-induced hypercalcemia (TIH), which has been similarly used as a model to determine the active doses of other bisphosphonates. The primary objective was to determine, with a dose escalation schedule, two nontoxic dose levels of zoledronate able to induce normocalcemia in at least 80% of patients with TIH after rehydration (corrected Ca for albumin levels >/=2.75 mmol/l). Based on estimates of potency, the starting dose was 0.002 mg/kg, and further tested doses were 0. 005, 0.01, 0.02, and 0.04 mg/kg. To obtain a more precise estimate of the response rate, we treated 10 more patients at the highest of the two effective dose levels. The median infusion time of zoledronate was 30 minutes. Thirty out of the 33 treated patients were evaluable for efficacy. Thirty percent of the patients had breast cancer and 54% had metastatic bone involvement. For all groups combined, mean Ca levels at baseline was 3.0 mmol/l. The two effective dose levels were 0.02 mg/kg and 0.04 mg/kg. Five out of five patients became normocalcemic after 0.02 mg of zoledronate/kg and 14 out of 15 after 0.04 mg of zoledronate/kg. The success rate of the latter dose was thus 93% (95% confidence interval [CI] 68-100%). At this dose, the first day of normocalcemia was day 2 or 3 for all but one patient. The duration of normocalcemia for the two effective doses could be assessed in nine patients; seven patients remained normocalcemic throughout the trial (32-39 days). The fall in serum Ca was accompanied by a marked fall in fasting urinary Ca excretion. Zoledronate was well tolerated: 7 out of 33 patients developed transient hypophosphatemia, and 3 developed transient hypocalcemia. The only clinically detectable side effect was an increase in body temperature occurring in 10 (30%) patients. In summary, very low doses of zoledronate (0.02 mg/kg and 0.04 mg/kg, i. e., 1.2 mg and 2.4 mg for a 60-kg individual, respectively) administered by a short-time infusion effectively treated patients with TIH. The fall in serum Ca was rapid, and normocalcemia was often maintained for several weeks. Zoledronate was well tolerated. Future trials will determine whether prolonged treatment with this potent compound can have greater effects on the skeletal morbidity rate in patients with tumor bone disease than can be achieved with currently available bisphosphonates. PMID- 10469285 TI - Development and characterization of a human in vitro resorption assay: demonstration of utility using novel antiresorptive agents. AB - A human in vitro resorption assay has been developed using osteoclastoma-derived osteoclasts and used to evaluate novel antiresorptive agents including antagonists of the alphavbeta3 integrin, and inhibitors of cathepsin K and the osteoclast ATPase. The potency of novel compounds in the in vitro resorption assay correlates with functional assays for each class of inhibitor: the human alphavbeta3-mediated cell adhesion assay for the vitronectin receptor antagonists (r2 = 0.82), the chick osteoclast vacuolar ATPase enzyme assay for the H+-ATPase inhibitors (r2 = 0.77) and the recombinant human cathepsin K enzyme assay for the cathepsin K inhibitors (r2 = 0.80). Cell suspensions, rich in osteoclasts, are prepared by collagenase digestion of the tumor tissue. These cells can be stored long-term in liquid nitrogen and upon thawing maintain their bone-resorbing phenotype. The cryopreserved cells can be cultured on bovine cortical bone for 24 48 h and resorption can be measured by either confocal microscopy or biochemical assays. The resorptive activity of osteoclasts derived from a number of tumors can be inhibited reproducibly using a number of mechanistically unique antiresorptive compounds. In addition, the measurement of resorption pits by laser confocal microscopy correlates with the release of type I collagen C telopeptides or N-telopeptides, as measured by enzyme-linked immunosorbent assay. Resorption can be measured reproducibly using a 48-h incubation of osteoclasts on bone slices, or a 24-h incubation with bone particles. This in vitro human osteoclast resorption assay provides a robust system for the evaluation of inhibitors of osteoclastic function that may be developed for the treatment of metabolic bone diseases such as osteoporosis. PMID- 10469286 TI - Low cell motility induced by hsp27 overexpression decreases osteolytic bone metastases of human breast cancer cells in vivo. AB - The mechanisms controlling the formation of osteolytic bone metastases in patients with breast cancer are still poorly understood. To explore the role of motility in the establishment of osteolytic bone metastases, we have used a model of bone metastasis in which MDA-MB-231 breast cancer cells exhibiting low (hsp27 transfectants) and high (control-transfectant) endogenous cell motility were compared. We found that MDA-MB-231 cells exhibiting low cell motility were less capable of establishing osteolytic lesions. The number and the area of the osteolytic lesions in mice inoculated with low motility cells were both significantly smaller. Histomorphometry of bone lesions also demonstrated less tumor area in mice bearing hsp27 transfectants although there was no difference in the osteoclast number per square millimeter of tumor-bone interface. These data suggest that cell motility may be an important mechanism in the metastatic cascade of breast cancer cells to the bone and that controlling cell motility may be a useful target to prevent the establishment of osteolytic bone metastases. PMID- 10469287 TI - Lack of association between estrogen receptor genotypes and bone mineral density, fracture history, or muscle strength in elderly women. AB - The PvuII polymorphism of the estrogen receptor (ESR) gene and its relation to bone mineral density (BMD), fracture history, and muscle strength was studied in 313 postmenopausal (76 +/- 5 years) women of Caucasian origin, of whom 142 had suffered from a fragility fracture after the age of 50 years (14 with fracture of the hip, 38 of the spine, 45 of the wrist, and 85 of other bones). The ESR genotype distribution was similar in women with and without a history of fragility fracture (PP 21%, Pp 43%, pp 36% compared with PP 18%, Pp 47%, pp 35%). We did not find a correlation between the ESR genotypes and BMD at the lumbar spine, the femoral neck, or the proximal forearm. No association was found with grip or quadriceps strength. We further evaluated the relationship between the vitamin D receptor (VDR) and ESR haplotypes and BMD in a random subgroup of 270 elderly women. No differences were found in women with the BBpp versus the bbPP haplotype in the femoral neck (mean difference +/- SD, in Bbpp compared with bbPP groups: -0.05 +/- 0.15 g/cm2), the spine (0.01 +/- 0.13 g/cm2), or the forearm (0.04 +/- 0.08 g/cm2). The significant association of quadriceps strength with VDR genotypes (25% lower in BB compared with bb genotype, p < 0.05) was not influenced by ESR haplotypes. We conclude that in elderly Caucasian women the PvuII ESR polymorphism is not associated with osteoporosis, fracture history, nor muscle strength and does not influence the association of bone density and muscle strength with polymorphism of the VDR. PMID- 10469288 TI - The relationship of biochemical markers of bone turnover to bone density changes in postmenopausal women: results from the Postmenopausal Estrogen/Progestin Interventions (PEPI) trial. AB - We assessed the associations of eight bone turnover markers (BTMs) with baseline and 1-year percentage changes in lumbar spine and hip bone mineral density (BMD) of 293 postmenopausal women undergoing treatment with hormone replacement therapy (HRT) or placebo using squared correlation coefficients (R2). In 239 women assigned to treatment with estrogen alone or with with estrogen plus progestins (active treatment), mean percentage changes for all markers decreased significantly and remained below baseline values through 3 years of study, whereas mean percentage changes for 54 women assigned to the placebo group showed no significant change from baseline in any marker. At baseline, age and body mass index (BMI) together accounted for 16% and 25% of the variance in spine and hip BMD, respectively. The telopeptide resorption marker, cross-linked N-telopeptide of type I collagen (NTX), alone accounted for 12% and 8% of variance, respectively. Another telopeptide, carboxy-terminal telopeptide of type I collagen (Crosslaps), accounted for 8% and 7% of variance, respectively. A bone specific alkaline phosphatase (BALP-2) accounted for 8% of variance at the spine and 5% at the hip. No other marker accounted for more than 5% of total variance at either site; adding either baseline NTX, Crosslaps, or BAP-2 to regressions containing age and BMI increased R2 values at the spine and hip to about 22% and 28%, respectively. In the placebo group, baseline spine BMD accounted for 4% of the variance in 1-year spine BMD percentage change, whereas baseline values for age and BMI accounted for 1% and 0% of the variance, respectively; none of the three accounted for more than 0% of hip BMD percentage change; Crosslaps and NTX contributed 5% and 4% to the variance in 1-year spine BMD percentage change, but other markers accounted for < 2% of variance at the spine. At the hip, another BALP (BALP-1) accounted for 4% of variance, but no other baseline marker except NTX accounted for more than 1% of variance. In the active treatment group, baseline values for age, BMI, and spine BMD together accounted for 13% of the percentage change in spine BMD and for 4% of the BMD change at the hip. No individual or pair of baseline markers significantly enhanced these R2 values, but addition of 1-year percentage changes in some individual markers did significantly increase it. The largest R2 value was obtained by adding the percentage change in BALP-2, which increased the R2 in spine BMD percentage change to 20% and that at the hip to 8%. Adding baseline and change variables for all eight markers to the regression increased R2 to 28% at the spine and 12% at the hip. Restricting the set of analyses to individuals who suppressed marker activity beyond the precision error for the measurement did not improve R2s for the regressions. When baseline marker values were stratified into quartiles, only NTX and osteocalcin showed significant relationships between quartile and change in spine BMD, and these did not reach significance at the hip. When the 1-year change in markers was stratified into quartiles, significant relationships with percentage change in spine BMD were observed only for BALP phosphatases. We conclude that BTMs are not a surrogate for BMD to identify women with low bone mass and that they offer little useful information for predicting BMD changes for individual untreated or HRT-treated postmenopausal women. PMID- 10469289 TI - Trabecular bone turnover and bone marrow cell development in tail-suspended mice. AB - To clarify the relationship between the changes of trabecular bone turnover and bone marrow cell development during mechanical unloading and reloading, we performed experiments with tail-suspended mice. At 8 weeks of age, 150 male ddY mice were divided into three body weight-matched groups. Mice of group 1 were euthanized at the start of tail suspension (day 0) as a baseline control. The mice of group 2 were subjected to hindlimb unloading by tail suspension for 14 days and reloading for the subsequent 14 days. The mice of group 3 were normally loaded as age-matched controls. Mice of groups 2 and 3 were sacrificed at 7, 14, and 28 days after the start of the experiment. In the first experiment (histomorphometric study of tibiae), unloading for 7 and 14 days and reloading for the subsequent 14 days significantly decreased the bone volume compared with that in the age-matched controls, respectively. Unloading for 7 and 14 days also significantly reduced the bone formation rate (BFR/BS), respectively, but reloading for the subsequent 14 days restored BFR/BS to the control level. While the unloading for 7 and 14 days significantly increased both the osteoclast surface (Oc.S/BS) and the osteoclast number (Oc.N/BS), the reloading for the subsequent 14 days decreased Oc.S/BS and Oc. N/BS, respectively. In the second experiment (bone marrow cell culture study of tibiae), unloading for 7 and 14 days reduced the adherent stromal cell number, without significance. Unloading for 7 days significantly decreased the mineralized nodule formation. Reloading for the subsequent 14 days markedly increased the adherent stromal cell number and the mineralized nodule formation. Unloading for 7 days significantly increased the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells. These data clearly demonstrate that unloading reduces bone formation and increases bone resorption, and subsequent reloading restores reduced bone formation and suppresses increased bone resorption, closely associated with the changes in adherent stromal cell number, mineralized nodule formation, and the number of TRAP-positive multinucleated cells. PMID- 10469290 TI - Morphometric dual-energy X-ray absorptiometry of the spine: report of a large series and correlation with axial bone mineral density. AB - We studied vertebral morphometry and its relation to bone mineral density (BMD) in normal Brazilian women (n = 605). All women (age 22-97 years) were ambulatory and healthy. A lateral spine scan was done for morphometric X-ray absorptiometry using an imaging densitometer. In 429 of these women, BMD of the spine and proximal femur also were measured using dual-energy X-ray absorptiometry. All women were white with mean (+/- 1 SD) age of 53.7 (+/- 9.5) years. About 21% of the women over 50 years had a T score for spine BMD lower than -2.5 SD, and 7% had a femoral neck BMD below this osteoporosis threshold. Vertebral heights (anterior, HA; middle, HM; and posterior, HP) and ratios (HA/HP and HM/HP) were assessed. There was no systematic difference between younger (20-49 years) and older (50+ years) women in heights or ratios. The vertebral heights were normalized for those observed in each individual case for the L2-L4 sequence. This normalization was adequate for all vertebral heights; the Z score averaged about +0.1. The average Z score for HA/HP was +0.01, but that for the HM/HP was 0.72, indicating that the latter ratio might differ from the reference population used (white American and European women). We observed a small positive correlation between vertebral heights and spine or femur BMD, but this was due entirely to the influence of body size on BMD. On a group basis, the HM/HP showed a significant association with axial BMD; the 1 SD difference between the lowest and highest quartile was associated with a difference of 8-15% (0.5-1.0 SD) in axial BMD. PMID- 10469291 TI - Markers of bone turnover predict postmenopausal forearm bone loss over 4 years: the OFELY study. AB - The ability of biochemical markers to predict the rate of postmenopausal bone loss is still controversial. To investigate this issue further, baseline levels of a panel of specific and sensitive biochemical bone markers were correlated to the rate of change of forearm bone mineral density (BMD) assessed by four measurements over a 4-year period using dual-energy X-ray absorptiometry in a large population-based prospective cohort of 305 women aged 50-88 years (mean 64 years), 1-38 years postmenopausal. In the whole population, higher baseline levels of bone formation (serum osteocalcin and serum type I collagen N-terminal propeptide) and bone resorption markers (urinary N-telopeptides; urinary and serum C-telopeptides) were significantly associated with faster BMD loss (r = 0.19 to -0.30, p < 0.001), independently of age. In women within 5 years of menopause that have the highest rate of bone loss, the predictive value of bone markers was increased with correlation coefficients reaching 0.53. Women with an abnormally high bone turnover, i.e., with levels of bone markers at baseline 2 SD above the mean of premenopausal women, had a rate of bone loss that was 2- to 6 fold higher than women with a low turnover (p = 0.01-0.0001) according to the marker. When the population was categorized according to quartiles of bone markers at baseline, a similar relationship between increased levels of bone markers and faster rate of bone loss was found (p = 0.008-0.0001). In the logistic regression model, the odds-ratio of fast bone loss, defined as the rate of bone loss in the upper tertile of the population, was increased by 1.8- to 3.2 fold for levels of biochemical markers in the high turnover group compared with levels within the premenopausal range, with, however, a limited value for identifying individual fast bone losers. We conclude that increased levels of some of the new biochemical markers of bone turnover are associated with greater radial bone loss. Because increased bone loss is associated with an increased risk of fracture, bone turnover markers may be useful to improve the prediction of the risk of osteoporosis in postmenopausal women. PMID- 10469292 TI - Low body mass index is an important risk factor for low bone mass and increased bone loss in early postmenopausal women. Early Postmenopausal Intervention Cohort (EPIC) study group. AB - Thinness (low percentage of body fat, low body mass index [BMI], or low body weight) was evaluated as a risk factor for low bone mineral density (BMD) or increased bone loss in a randomized trial of alendronate for prevention of osteoporosis in recently postmenopausal women with normal bone mass (n = 1609). The 2-year data from the placebo group were used (n = 417). Percentage of body fat, BMI, and body weight were correlated with baseline BMD (r = -0. 13 to -0.43, p < 0.01) and 2-year bone loss (r = -0.14 to -0.19, p < 0.01). Women in the lowest tertiles of percentage of body fat or BMI had up to 12% lower BMD at baseline and a more than 2-fold higher 2-year bone loss as compared with women in the highest tertiles (p A polymorphism at position -238 of the tumor necrosis factor-alpha promoter (TNF alpha-238*A allele) was more common in type I psoriasis (27%) than in the controls [9.8%; odds ratio 3.4 (95% confidence interval 1.6-7.2); p = 0.0012; pcorr = 0.018]. Surprisingly, this overrepresentation of the tumor necrosis factor alpha-238*A allele was observed in male patients [4.1 (1.5-11.0); p = 0.0046; pcorr = 0.064] but not in female patients [1.8 (0.5-6.5); p = 0.5]. The G-->A polymorphism at position -308 of the tumor necrosis factor-alpha promoter was less frequent in type I psoriasis (23%) compared with controls (35.7%), although the negative association was weak [0.54 (0.3-0.97); p = 0.041; pcorr = not significant]. The distribution of the TNF alpha-238*A and TNF alpha-238*A alleles was similar in type II patients and controls. Our results suggest that male carriers of the G-->A polymorphism at position -238 of the tumor necrosis factor-alpha promoter are at an increased risk to develop early-onset psoriasis. PMID- 10469307 TI - Low-intensity laser therapy is an effective treatment for recurrent herpes simplex infection. Results from a randomized double-blind placebo-controlled study. AB - Recurrent infection with herpes simplex virus is a common disease. Recently, alternative therapies have been introduced. Among those, low-intensity laser therapy mainly used for the acceleration of wound healing and in pain therapy has previously been shown to be of benefit in herpes zoster infections. In this study we evaluated the influence of low-intensity laser therapy (wavelength 690 nm, intensity: 80 mW per cm2, dose: 48 J per cm2) in 50 patients with recurrent perioral herpes simplex infection (at least once per month for more than 6 mo) in a randomized, double-blind placebo-controlled trial design. Patients in the laser group received daily irradiations for 2 wk, whereas patients in the placebo group were sham-irradiated. After completion of the laser/sham treatment, patients were asked to return to the Department of Dermatology, University of Vienna Medical School at the time of recurrence. All except two patients completed the study and were monitored for 52 wk. The median recurrence-free interval in the laser treated group was 37.5 wk (range: 2-52 wk) and in the placebo group 3 wk (range: 1-20 wk). This difference was found to be statistically significant (p < 0.0001; Wilcoxon's Rank Sum Test). In conclusion, we demonstrated that a total of 10 irradiations with low-intensity laser therapy significantly lowers the incidence of local recurrence of herpes simplex infection. Since this athermic phototherapeutic modality represents a safe, noninvasive treatment, it might be considered as an alternative to established therapeutic regimens in this indication. PMID- 10469308 TI - Genetic mapping and physical cloning of UVB susceptibility region in mice. AB - One of the most important goals of cancer research is to identify environmental and host factors that contribute to the malignant state. Human skin cancers are among the few tumor types for which the predominant environmental carcinogen is known. Ultraviolet light, a component of sunlight, is an important cause of skin cancer in humans. In humans and mice, ultraviolet B radiation induces systematic and local immunosuppression. A consequence of that is inappropriate immune surveillance of somatic tissues for evidence of malignantly transformed cells. The impairment of contact hypersensitivity, as it develops early and correlates well with tumor frequency in various mouse strains, has been used for over 15 y as a model of immunologic events occurring in photocarcinogenesis. In mice, as well as in humans, ultraviolet B radiation induced impairment of contact hypersensitivity is not uniform in all individuals; some individuals are susceptible to the deleterious effects of ultraviolet B, whereas others are resistant to ultraviolet B. We have defined the genetic locus responsible for ultraviolet B susceptibility and resistance in mice within the Bat5 and H-2D segment of the mouse chromosome 17. PMID- 10469309 TI - Epidermal dysplasia and abnormal hair follicles in transgenic mice overexpressing homeobox gene MSX-2. AB - The homeobox gene Msx-2 is expressed specifically in sites of skin appendage formation. To explore its part in skin morphogenesis, we produced transgenic mice expressing Msx-2 under the control of the cytomegalovirus promoter. The skin of these transgenic mice was flaky, exhibiting desquamation and shorter hairs. Histologic analysis showed thickened epidermis with hyperproliferation, which was restricted to the basal layer. Hyperkeratosis was also evident. A wide zone of suprabasal cells were misaligned and coexpressed keratins 14 and 10. There was reduced expression of integrin beta 1 and DCC in the basal layer. Hair follicles were misaligned with a shrunken matrix region. The dermis showed increased cellularity and empty vacuoles. We suggest that Msx-2 is involved in the growth control of skin and skin appendages. PMID- 10469310 TI - Expression and role of IL-15 in post-burn hypertrophic scars. AB - Hypertrophic scarring is a skin disorder that occurs after wounding and thermal injury. There is accumulating evidence that immunologic processes such as infiltration of activated T lymphocytes and altered cytokine production may play a role in the formation of hypertrophic scars. Interleukin-15, a cytokine identified as a T cell growth factor, also acts as a chemoattractant for T cells and has pro-inflammatory properties. We investigated the expression and the role of this cytokine in hypertrophic scarring. IL-15 expression was compared in skin biopsies of hypertrophic scars (HS) both in active (AHS) and in remission (RHS) phases, in normotrophic scars (NTS) and in normal skin using reverse transcriptase-polymerase chain reaction and immunohistochemistry. IL-15 expression in HS was significantly higher than in NTS or normal skin. Furthermore, AHS expressed higher levels of IL-15 than RHS. Immunohistologic analysis of AHS samples showed strong IL-15 immunoreactivity in keratinocytes and Langerhans cells in the epidermis and in macrophages, fibroblasts, and dermal dendritic cells in the dermis. High levels of IL-15 expression in AHS correlated with abundant infiltration of activated CD3+ cells. Ex vivo experiments indicate that IL-15 can sustain the proliferative response of T cells derived from AHS but not from RHS and NTS. In addition, IL-15 prevents both cytokine deprivation and activation-induced apoptosis of T cells derived from AHS. Taken together, these results suggest that IL-15 can be involved in the recruitment, proliferation, and apoptosis inhibition of T cells in AHS. The findings that the evolution from an AHS to a RHS is associated with a decrease in IL15 expression, and with a loss of IL-15 responsiveness in ex vivo-cultured T cells, indicate that this cytokine plays an important role in the biology of pathologic scar formation. PMID- 10469311 TI - Expression of type XVI collagen in human skin fibroblasts: enhanced expression in fibrotic skin diseases. AB - Abundance of type XVI collagen mRNA in normal human dermal fibroblasts explanted from different horizontal layers was determined using RNase protection assays. Type XVI collagen mRNA level in the fibroblasts explanted from the upper dermis was greater than those of the middle and lower dermis. The antibody raised against the synthetic N-terminal noncollagenous region reacted with approximately 210 kDa collagenous polypeptide in the culture medium of fibroblasts. Immunohistochemical study of normal human skin demonstrated that the antibody reacted preferentially with the fibroblasts and the extracellular matrix in the upper dermis rather than those in the middle and lower dermis. Type XVI collagen mRNA level was elevated 2.3-fold in localized scleroderma and 3.6-fold in systemic scleroderma compared with keloid and normal controls. Immunofluorescent study revealed that an intense immunoreactivity with the antibody was observed in the upper to lower dermal matrix and fibroblasts in the skin of systemic scleroderma as compared with normal skin. The results suggest that expression of type XVI collagen, a member of fibril-associated collagens with interrupted triple helices, in human skin fibroblasts can be heterogeneous in the dermal layers and can be modulated by some fibrotic diseases. PMID- 10469312 TI - A newly identified patient with clinical xeroderma pigmentosum phenotype has a non-sense mutation in the DDB2 gene and incomplete repair in (6-4) photoproducts. AB - We report here a patient (Ops1) with clinical photosensitivity, including pigmented or depigmented macules and patches, and multiple skin neoplasias (malignant melanomas, basal cell carcinomas, and squamous cell carcinomas in situ) in sun-exposed areas. These clinical features are reminiscent of xeroderma pigmentosum. As cells from Ops1 showed normal levels in DNA repair synthesis in vivo (unscheduled DNA synthesis and recovery of RNA synthesis after ultraviolet irradiation), we performed a postreplication repair assay and recovery of replicative DNA synthesis after ultraviolet irradiation to investigate if Ops1 cells belonged to a xeroderma pigmentosum variant pattern. Ops1 cells were normal, but there was an incomplete pattern repair in (6-4) photoproducts in contrast to a normal pattern repair in cis-syn cyclobutane pyrimidine dimers by repair kinetics using the enzyme-linked immunosorbent assay. Moreover, Ops1 cells were defective in a damage-specific DNA binding protein and carried a non-sense mutation in the DDB2 gene. These results suggest that (i) the DDB2 gene is somewhat related to skin carcinogenesis, photoaging skin, and the removal of (6 4) photoproducts; (ii) although it is believed that cyclobutane pyrimidine dimers are the principal mutagenic lesion and (6-4) photoproducts are less likely to contribute to ultraviolet-induced mutations in mammals, Ops1 is one of the ultraviolet-induced mutagenic models induced by (6-4) photoproducts. PMID- 10469313 TI - erbB-2 overexpression but no activation of beta-Catenin gene in extramammary Paget's disease. AB - Our previous study in extramammary Paget's disease showed neither p53 mutations nor allelic loss at selected loci implicated in other cancers, suggesting a pathogenesis of this skin cancer different from other common epithelial malignancies. To examine further the genetic defects in extramammary Paget's disease, we carried out molecular genetic analyses in 31 tumor samples obtained from 27 cases of extramammary Paget's disease without underlying malignancies. Immunohistochemistry using CB-11 monoclonal antibody revealed either membrane or cytoplasmic erbB-2 oncoprotein overexpression in none of the 13 primary in situ tumors, but in one recurrent in situ tumor, 10 of 13 invasive primary tumors and two of four lymph node metastases. Sensitive dual color fluorescence in situ hybridization analysis using probes for erbB-2 gene locus and chromosome 17 pericentromere, however, revealed different erbB-2 gene status in the erbB-2 overexpressing tumors. One recurrent in situ tumor and one lymph node metastasis showed definite gene amplification characterized by multiple scattered signals or a few large clustered erbB-2 signals, whereas four tumors with predominantly cytoplasmic erbB-2 overexpression were thought to have low-grade gene amplification. The remaining six tumors overexpressing erbB-2 showed no increase of erbB-2 copy numbers. No evidence of abnormal activation of the beta-catenin gene, a critical mediator of Wnt signaling pathway, in any tumor by immunohistochemical staining and by direct sequencing and reverse transcription polymerase chain reaction analysis was found. Frequent overexpression of erbB-2 by either gene amplification or possible transcriptional activation in invasive primary tumors and metastases suggests an important part for this oncogene in the progression of extramammary Paget's disease. PMID- 10469314 TI - Monilethrix: a novel mutation (Glu402Lys) in the helix termination motif and the first causative mutation (Asn114Asp) in the helix initiation motif of the type II hair keratin hHb6. AB - Monilethrix, a rare human hair disorder with autosomal dominant transmission, can be caused by mutations in hair keratins. Up to now, causative mutations have only been found in two type II cortex keratins, hHb6 and hHb1. In these hair keratins, the helix termination motif, HTM, was the only site in which mutations were located. The most frequent mutation, which has been found in 22 cases, was a Glu413Lys substitution in hHb6, whereas other mutations, i.e., hHb6 Glu413Asp, hHb1 Glu413Lys, and hHb1 Glu402Lys, have been reported in a distinctly lower number of cases. In this study, we describe the equivalent of the hHb1 Glu402Lys mutation in the HTM of cortex keratin hHb6. The mutation occurred in an American family in which it could only be detected in one clinically affected individual. Thus the underlying G-->A transition represents a spontaneous germ-line mutation in the hHb6 gene. This new mutation indicates that both the hHb6/hHb1 Glu413Lys substitution and the hHb6/hHb1 Glu402Lys substitution, represent mutational hotspots in the HTM of type II cortex keratins. However, we also describe a monilethrix-causing mutation in the helix initiation motif, HIM, of the cortex keratin hHb6. The critical Asn114Asp substitution was only found in affected members of a large Swedish three-generation family. Considering that since childhood, half of the affected individuals suffer from complete baldness and follicular keratosis, the new HIM mutation seems to be associated with a rather severe disease phenotype. In conclusion, our data strongly suggest that monilethrix is a disease of the hair cortex, whose etiology is interesting in that causative mutations seem to be restricted to type II hair keratins. PMID- 10469315 TI - Identification of epitopes on tyrosinase which are recognized by autoantibodies from patients with vitiligo. AB - The identification of tyrosinase autoantibodies in some patients with vitiligo has previously been reported. In this study we have determined the B cell epitopes on tyrosinase which are recognized by these autoantibodies. Deletion derivatives of tyrosinase cDNA were constructed and then translated in vitro with the concomitant incorporation of [35S]methionine into the protein products. The 35S-labeled tyrosinase derivatives were subsequently used in radioimmunoassays to investigate the reactivity of sera from five vitiligo patients. The epitope regions identified were: three in a central region of tyrosinase (amino acids 240 255, 289-294, and 295-300) and two others towards the C-terminal end of the protein (amino acids 435-447 and 461-479). Computer analysis of the potential B cell epitopes on tyrosinase revealed that the epitope regions recognized by the vitiligo sera were located in areas predicted to be highly antigenic. In addition, the centrally located antigenic regions (amino acids 289-294 and 295 300) had amino acid sequence homology to both tyrosinase-related protein-1 and 2. Thus, the epitopes on tyrosinase recognized by vitiligo patient sera are heterogeneous and include a region with homology to two related proteins which may explain the cross-reactivity previously noted between these antigens. PMID- 10469317 TI - Serum androgens and genetic linkage analysis in early onset androgenetic alopecia. PMID- 10469316 TI - Von Willebrand factor receptor GPIb alpha is expressed by human factor XIIIa positive dermal dendrocytes and is upregulated by mast cell degranulation. AB - GPIb alpha, a glycoprotein component of the GPIb-IX-V complex, serves as a platelet membrane receptor that mediates adhesion to von Willebrand factor normally present in the vascular subendothelium. Recent data have demonstrated that GPIb alpha is not restricted to platelets, but is also expressed by endothelium in vitro. In this study, we describe the expression and distribution of GPIb alpha in normal adult and neonatal human skin. GPIb alpha is present, as detected by immunohistochemistry, on endothelial cells and on highly dendritic cells localized within the perivascular space, dermal-epidermal junction, and reticular dermis. By dual-labeling immunofluorescence and confocal microscopy, GPIb alpha-positive cells within the dermal interstitium are demonstrated to represent factor XIIIa-positive dermal dendrocytes. In organ cultures of neonatal human foreskin, mast cell degranulation induced by either substance P or compound 48/80 resulted in transiently increased GPIb alpha expression by dermal dendrocytes. Because the GPIb-IX-V complex plays a part in regulating hemostasis and may be important for cellular interactions with extracellular matrix molecules, these data provide additional insight into the potential function of FXIIIa-positive dermal dendrocytes in skin remodeling and repair. PMID- 10469318 TI - Basal cell carcinoma-like epidermal changes overlying dermatofibromas often reveal loss of heterozygosity in the PTCH gene. PMID- 10469319 TI - A homozygous nonsense mutation in the zinc-finger domain of the human hairless gene underlies congenital atrichia. PMID- 10469320 TI - UV induces p21WAF1/CIP1 protein in keratinocytes without p53. PMID- 10469321 TI - X-linked anhidrotic (hypohidrotic) ectodermal dysplasia caused by a novel mutation in EDA1 gene: 406T > G (Leu55Arg) PMID- 10469322 TI - What is the best way to define the antimelanocyte T cell repertoire? PMID- 10469323 TI - Human herpesvirus 7 and pityriasis rosea. PMID- 10469324 TI - In vivo confocal scanning laser microscopy of human skin II: advances in instrumentation and comparison with histology. AB - In 1995, we reported the construction of a video-rate scanning laser confocal microscope for imaging human skin in vivo. Since then, we have improved the resolution, contrast, depth of imaging, and field of view. Confocal images of human skin are shown with experimentally measured lateral resolution 0.5-1.0 microm and axial resolution (section thickness) 3-5 microm at near-infrared wavelengths of 830 nm and 1064 nm; this resolution compares well to that of histology which is based on typically 5 microm thin sections. Imaging is possible to maximum depth of 350 microm over field of view of 160-800 microm. A mechanical skin-contact device was developed to laterally stabilize the imaging site to within +/- 25 microm in the presence of subject motion. Based on these results, we built a small, portable, and robust confocal microscope that is capable of imaging normal and abnormal skin morphology and dynamic processes in vivo, in both laboratory and clinical settings. We report advances in confocal microscope instrumentation and methods, an optimum range of parameters, improved images of normal human skin, and comparison of confocal images with histology. PMID- 10469325 TI - Percutaneous penetration of local anesthetic bases: pharmacodynamic measurements. AB - Local anesthetics do not penetrate readily through human skin if applied in their salt form; however, if applied in their base form various effects may be observed, such as a decrease in pricking pain and a change in burning, itch, and thermal sensations. These effects occur after skin penetration and may be attributed to the action of the anesthetics on nociceptors and thermoreceptors, i.e., on C and Adelta nerve fiber respectively. As there is little known about the time course of the pharmacodynamic response of cutaneously applied local anesthetic bases, this study was conducted to characterize various local anesthetics pharmacodynamically by measuring thermal thresholds over time with a thermal sensory analyzer. The results show that the investigated local anesthetics affect thermal thresholds to a different extent, with tetracaine and lidocaine being most efficient. From the response versus time profiles of all eight study subjects various response parameters were obtained: only the cold sensation parameters proved suitable for characterization of the local anesthetics, possibly because cold receptors are located in the epidermis and can easily be reached. Lag times of onset are short and the maximum anesthetic effect is reached within 2-3 h. Cold sensation parameters correlate linearly with the solubility of the local anesthetic bases in medium chain triglycerides and with the drug flux of 50% saturation, indicating that medium chain triglycerides may have similar properties with regard to the local anesthetics solubility as the stratum corneum lipids. PMID- 10469326 TI - The mRNA for protease nexin-1 is expressed in human dermal papilla cells and its level is affected by androgen. AB - Protease nexin-1, an inhibitor of serine proteases, plays important parts in the regulation of the growth, differentiation, and death of cells by modulating proteolytic activity. The mRNA for protease nexin-1 accumulates in rat dermal papilla cells in a hair cycle-dependent fashion and its levels are well correlated with the ability of dermal papilla cells to support hair growth. In an attempt to characterize the potential role of protease nexin-1 as a modulator of hair growth in humans, we investigated the steady-state level of protease nexin-1 mRNA in cultured human dermal papilla cells using a semiquantitative technique that involved reverse transcription and polymerase chain reaction, as well as the localization of this mRNA in vivo using dissected hair follicles. Protease nexin 1 mRNA was expressed in all dermal papilla cells examined, and it was also identified in the lower part of the connective tissue sheath. Moreover, we found that levels of protease nexin-1 mRNA were depressed by dihydrotestosterone, the most potent androgen, in cultured dermal papilla cells obtained from balding scalp. Our results suggest that protease nexin-1 might be a key molecule in the control of hair growth in humans and, moreover, that the androgen-mediated downregulation of the synthesis of protease nexin-1 might be associated with the progression of male-pattern baldness. PMID- 10469327 TI - Moderation of phenotypic severity in dystrophic and junctional forms of epidermolysis bullosa through in-frame skipping of exons containing non-sense or frameshift mutations. AB - Non-sense mutations on both alleles of either the type VII collagen gene (COL7A1) or the genes encoding laminin 5 (LAMA3, LAMB3, or LAMC2) usually result in clinically severe forms of recessive dystrophic or junctional epidermolysis bullosa, respectively. In this study we assessed two unrelated families whose mutations in genomic DNA predicted severe recessive dystrophic epidermolysis bullosa or junctional epidermolysis bullosa phenotypes but in whom the manifestations were milder than expected. The recessive dystrophic epidermolysis bullosa patients had a homozygous single base-pair frameshift mutation in exon 19 of COL7A1 (2470insG). Clinically, there was generalized blistering but only mild scarring. Skin biopsy revealed positive type VII collagen immunoreactivity and recognizable anchoring fibrils. The junctional epidermolysis bullosa patients were compound heterozygotes for a frameshift/non-sense combination of mutations in exons 3 and 17 of LAMB3 (29insC/Q834X). These patients did not have the lethal form of junctional epidermolysis bullosa but, as adults, displayed the milder generalized atrophic benign epidermolysis bullosa variant. There was undetectable laminin 5 staining at the dermal-epidermal junction using an antibody to the beta3 chain, but faintly positive alpha3 and gamma2 chain labeling, and there was variable hypoplasia of hemidesmosomes. To explain the milder recessive dystrophic epidermolysis bullosa and junctional epidermolysis bullosa phenotypes in these families, reverse transcription-polymerase chain reaction, using RNA extracted from frozen skin, was able to provide evidence for some rescue of mutant mRNA transcripts with restoration of the open- reading frame. In the recessive dystrophic epidermolysis bullosa patients, transcripts containing in-frame skipping of exon 19 of COL7A1 in the cDNA were detected, and in the junctional epidermolysis bullosa patients transcripts with in-frame skipping of exon 17 of LAMB3 were identified. The truncated proteins encoded by these transcripts are expected to lack certain critical domains involved in cell-matrix attachment, but may still be able to contribute to adhesion thereby moderating the severity of the skin blistering. This study shows the limitations in predicting phenotype in epidermolysis bullosa solely based on mutation analysis of genomic DNA and emphasizes the importance of immunohistochemistry, electron microscopy, and mRNA assessment as parallel investigations. PMID- 10469328 TI - Characterization of the major susceptibility region for psoriasis at chromosome 6p21.3. AB - Psoriasis is a common inflammatory skin condition caused by genetic and environmental factors. Recent genome-wide linkage analyses have identified a locus encoding susceptibility to psoriasis and placed this gene in the 12 cM interval between markers D6S426 and D6S276 on chromosome 6p21.3. This is a broad region and encompasses the human major histocompatibility complex. We have sought to localize the susceptibility gene more precisely by exploiting the linkage, haplotype, and linkage disequilibrium information available through genotyping 118 affected sib pairs, their parents and other affected family members. A total of 14 highly polymorphic markers were genotyped, combining anonymous loci with the class I genes HLA-B and -C distributed across a genetic interval of approximately 14 cM including the entire major histocompatibility complex. Through the application of higher density mapping within the major histocompatibility complex, we identified those regions most commonly shared identical by descent in patients with psoriasis. Using the transmission disequilibrium test, we found significant evidence of linkage and allelic association across an interval defined by the markers tn62 (p = 1.0 x 10(-7)), HLA-B (p = 4.0 x 10(-7)), and HLA-C (p = 2.7 x 10(-9)), a region encompassed within a 285 kb genomic DNA fragment. Hence these studies contribute to the refinement of the localization of a major psoriasis susceptibility gene and place the critical region near to HLA-C. PMID- 10469329 TI - Normal ultrastructure, but altered stratum corneum lipid and protein composition in a mouse model for epidermolytic hyperkeratosis. AB - Recently, we established keratin 10-deficient mice, serving as a model for the hyperkeratotic skin disorder epidermolytic hyperkeratosis. The considerable ichthyosis in these mice suggested alterations in terminal differentiation and in the formation of a functional epidermal barrier. Here, we report on the ultrastructural organization and composition of the stratum corneum lipids and on the expression of two major cornified envelope proteins. Electron microscopy of ruthenium tetroxide postfixed skin samples demonstrated a normal extrusion and morphology of lamellar bodies as well as the formation of bona fide lamellar layers in neonatal keratin 10-deficient mice. When we studied the composition of the major stratum corneum lipids, however, we found significant changes. Most importantly, the analysis of ceramide subpopulations revealed that the total amount of ceramide 2 was elevated in keratin 10-deficient mice, whereas ceramides 1, 3, 4, and 5 were decreased among total stratum corneum lipids. The amount of the ceramide precursors sphingomyelin and glucosylceramide was reduced in the stratum corneum without accompanying changes in the mRNA coding for acid sphingomyelinase. Notably, we found an increased mRNA and protein content for involucrin in neonatal keratin 10-deficient mice, whereas the expression of loricrin was not changed. Our data demonstrate that, although the formation of lipid layers in the stratum corneum appeared to be normal, its lipid composition is significantly altered in keratin 10-deficient mice. PMID- 10469330 TI - Protein oxidation in human stratum corneum: susceptibility of keratins to oxidation in vitro and presence of a keratin oxidation gradient in vivo. AB - The stratum corneum is located at the interface between body and environment and thus is constantly exposed to a pro-oxidative environment. Previously, we have demonstrated that stratum corneum lipids are targets of oxidative stress induced by ozone and by ultraviolet A and B exposure. Here, we employed an immunoblotting technique to detect protein oxidation in human stratum corneum obtained by tape stripping. After lysis, protein carbonyl groups were measured by derivatization with dinitrophenylhydrazine, separation by sodium dodecylsulfate-polyacrylamide gel electrophoresis, and immunoblotting using antibodies against dinitrophenyl groups. Keratin 10, identified by use of specific antibodies and by microsequencing, was demonstrated in vitro to be oxidizable by ultraviolet A irradiation, hypochlorite, and benzoyl peroxide. In vivo, a keratin 10 oxidation gradient with low levels in the lower stratum corneum layers, and about 3-fold higher contents of carbonyl groups towards the outer layers was demonstrated in forehead stratum corneum of healthy volunteers (n = 6). As protein oxidation can be associated with an increased susceptibility to proteases, this finding may be important for better understanding the process of desquamation. PMID- 10469332 TI - The time-course of psoralen ultraviolet A (PUVA) erythema. AB - The time-course for the development of ultraviolet A-induced erythema in psoralen sensitized skin differs from that caused by ultraviolet B or ultraviolet A but objective data are not available. During psoralen ultraviolet A therapy, the minimal phototoxic dose is determined 72 h after exposure, when psoralen ultraviolet A erythema is assumed to be maximal. This measurement is of fundamental importance in optimizing the therapeutic regimen. We examined a detailed time-course for development of psoralen ultraviolet A erythema in 16 subjects. The erythemal responses to ultraviolet B, ultraviolet A and psoralen ultraviolet A were assessed visually and using a reflectance device. Ultraviolet B erythema was maximal 24 h after exposure compared with subsequent time-points. Psoralen ultraviolet A erythema was evident at 24 h, with reduction in the median ultraviolet A minimal erythema dose from 14 to 5 J per cm2 in the presence of psoralen (p < 0.01; n = 9). Peak psoralen ultraviolet A erythema, assessed by minimal phototoxic dose, did not occur until 96 h or later in 75% of subjects. Using individual dose- response curves, we determined that only 67% of mean maximum psoralen ultraviolet A erythemal intensity had developed by 72 h. Furthermore, at the time of maximal erythema, the slope of the psoralen ultraviolet A dose-response curve was approximately 2-fold shallower than that for ultraviolet B-induced erythema. If assessment of psoralen ultraviolet A erythemal sensitivity had been made at 96 h instead of the conventional 72 h time point, peak erythemal responses would not have been missed in any of the subjects. Based on these findings, it seems appropriate to consider whether psoralen ultraviolet A minimal phototoxic dose measurements should be performed 96 h after exposure. PMID- 10469331 TI - Effects of a calcium receptor activator on the cellular response to calcium in human keratinocytes. AB - Changes in the concentration of extracellular calcium affect the balance between proliferation and differentiation in epidermal keratinocytes. Undifferentiated keratinocytes respond to the acute increase in the concentration of extracellular calcium with an increase of intracellular calcium concentration and inositol trisphosphate production, and, subsequently, the expression of differentiation related genes. Our previous studies demonstrated the presence of a calcium sensing receptor in human keratinocytes, which is identical to the parathyroid calcium-sensing receptor. In this study we showed that the calcimimetic compound NPS R-467, a selective calcium-sensing receptor activator, augmented the calcium elicited inositol trisphosphate response of cloned human keratinocyte calcium sensing receptor expressed in human embryonic kidney cells 293. In order to define the role of the calcium-sensing receptor in calcium induced epidermal differentiation, we investigated the ability of NPS R-467 to raise intracellular Ca2+ and stimulate differentiation in normal human foreskin keratinocytes. In the presence of 0.03 mM Ca2+, NPS R-467 increased the intracellular calcium concentration response in a concentration-dependent fashion. Undifferentiated normal human foreskin keratinocyte cells responded to increased extracellular calcium concentration with increased intracellular calcium concentration. NPS R 467 potentiated this response by increasing the maximal response. Its stereoisomer, NPS S-467, was not active in raising intracellular calcium concentration. Increasing extracellular calcium concentration from 0.03 to 1.2 mM stimulated the promoter activity of the differentiation marker gene, involucrin. NPS R-467 potentiated the calcium-stimulated increase in involucrin promoter activity unlike NPS S-467 or vehicle. Northern analysis of the normal human foreskin keratinocyte cells treated with NPS R-467 demonstrated potentiation of the calcium-stimulated increases in involucrin and transglutaminase mRNA levels. These results support the hypothesis that the calcium-sensing receptor expressed in keratinocytes mediates at least part of the intracellular calcium response to extracellular calcium and calcium-induced differentiation. PMID- 10469333 TI - Interleukins 18 and 12 synergistically upregulate interferon-gamma production by murine dendritic epidermal T cells. AB - A unique subset of gammadelta T cells, termed dendritic epidermal T cells, reside in murine epidermis. It was previously reported that freshly isolated dendritic epidermal T cells and dendritic epidermal T cell lines expressed mRNA for interferon-gamma. Recent studies indicated that interleukin-18, a novel cytokine which strongly induces interferon-gamma production by T cells, was produced by murine keratinocytes and Langerhans cells. Interleukin-12, which is regarded as a key cytokine for Th1 type helper clone responses, has also been reported to be produced by these cells in murine skin. In this study, we demonstrated by enzyme linked immunosorbent assay that interleukin-18 and interleukin-12 synergistically upregulated interferon-gamma production by dendritic epidermal T cells in short term cultures. This was the case in both C57/BL6 mice and BALB/C mice, although the quantity of interferon-gamma produced was different in the two mouse strains. Interleukin-18 or interleukin-12 alone did not induce interferon-gamma production by dendritic epidermal T cells. Interferon-gamma mRNA was only weakly detected by the semiquantitative reverse transcriptase-polymerase chain reaction method in freshly isolated dendritic epidermal T cells, and the mRNA expression was much increased 12 h after stimulation with interleukin-18 and interleukin-12. We also confirmed biologic activity of interferon-gamma produced by dendritic epidermal T cells by showing upregulation of major histocompatibility complex class II expression on Pam 212, murine keratinocyte cell line. Thus, this study suggests that interleukin-18 and interleukin-12 produced by keratinocytes and Langerhans cells regulate interferon-gamma production by dendritic epidermal T cells and thus may play important parts in the regulation of immune responses in skin associated lymphoid tissues. PMID- 10469334 TI - T cell stimulating stratum corneum antigens: characterization by chromatography and electrophoresis indicates limited diversity. AB - As part of a search for T cell autoantigens in inflammatory skin diseases, we have demonstrated proteinase K sensitive, denaturation stable, T cell stimulatory material with antigenic properties in aqueous extracts of stratum corneum from normal human skin. Activity was also demonstrable in extracts of whole epidermis. A combination of preparative, analytical, and microbore reversed phase high performance liquid chromatography, chromatofocusing, and denaturing preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis indicated limited structural diversity. Five components were separated, with Mr values from 5 to 18 kDa and apparent PI values from 4.5 to 10. Three components were purified to near homogeneity and showed molecular weights of 5, 13.5, and 18 kDa. Their potency was shown by the ability to induce stimulation indices of 20-89 with peripheral blood mononuclear cells and >500 with T cell lines. Use of inhibitors indicated that the active materials were not generated by the in vitro actions of proteases during extraction. The five partially purified components induced a time course of peripheral blood mononuclear cell proliferation compatible with the effects of antigen rather than superantigen. The 5 kDa component was rigorously bulk purified to yield a fraction that induced potent T cell activation but contained minimal detectable protein, a further indication of its biologic potency. Normal stratum corneum thus contains previously undescribed T cell antigens of high potency but limited structural diversity. The present data form a basis for determining their structure, cellular origin, and pathogenic relevance. PMID- 10469335 TI - Thymosin beta4 accelerates wound healing. AB - Angiogenesis is an essential step in the repair process that occurs after injury. In this study, we investigated whether the angiogenic thymic peptide thymosin beta4 (Tbeta4) enhanced wound healing in a rat full thickness wound model. Addition of Tbeta4 topically or intraperitoneally increased reepithelialization by 42% over saline controls at 4 d and by as much as 61% at 7 d post-wounding. Treated wounds also contracted at least 11% more than controls by day 7. Increased collagen deposition and angiogenesis were observed in the treated wounds. We also found that Tbeta4 stimulated keratinocyte migration in the Boyden chamber assay. After 4-5 h, migration was stimulated 2-3-fold over migration with medium alone when as little as 10 pg of Tbeta4 was added to the assay. These results suggest that Tbeta4 is a potent wound healing factor with multiple activities that may be useful in the clinic. PMID- 10469336 TI - Counterregulation of interleukin-18 mRNA and protein expression during cutaneous wound repair in mice. AB - Recent work has suggested interleukin-18 to represent a proinflammatory cytokine that contributes to systemic and local inflammation. As the process of cutaneous wound healing crucially involves an inflammatory phase of repair, we investigated the regulation of interleukin-18 during the repair process. In non-wounded skin we observed high levels of interleukin-18 mRNA, whereas corresponding interleukin 18 protein was expressed only at low basal levels. Upon injury, we found a rapid and large induction of interleukin-18 protein expression, which is directly correlated with decreasing mRNA levels within the wound. Immunohistochemical analysis revealed different sites of expression in the wounded area, with keratinocytes as one major source of interleukin-18 production. The counterregulation of interleukin-18 mRNA and protein expression during wound repair in vivo might represent a general mechanism for interleukin-18 expressional regulation, as cytokine-stimulated keratinocytes exhibit a similar downregulation of interleukin-18 mRNA that is directly associated with increasing interleukin-18 protein levels in vitro. The rapid induction of interleukin-18 during wound healing suggests a role for interleukin-18 within the early phase of repair rather than a role in costimulation of interferon-gamma release from T cells, which are present in high numbers within the wounded area only during the late inflammatory phase of repair. PMID- 10469337 TI - Adenoviral-mediated overexpression of platelet-derived growth factor-B corrects ischemic impaired wound healing. AB - Chronic wounds represent a major clinical problem with significant morbidity and healthcare expenditures, but no effective therapies. Topical platelet-derived growth factor-BB trials have required large and repeated doses to achieve only a modest effect. We examined the ability of an adenovirus containing the platelet derived growth factor-B transgene to improve the rate of wound healing through induction of platelet-derived growth factor-B overexpression in cells participating in the wound healing response. We treated excisional wounds in the ischemic rabbit ear, which have a 60% delay in healing, with vehicle, 106, or 108 plaque-forming units of an adenovirus containing the platelet-derived growth factor-B per wound (n = 19). At 7 d this resulted in a decrease in the epithelial gap from 3.4 +/- 1 mm (mean +/- SD) in vehicle-treated wounds to 1.9 +/- 1.8 mm (mean +/- SD, p < 0.05) when treated with 106 plaque-forming units of an adenovirus containing the platelet-derived growth factor-B, and 0.7 +/- 1.1 mm (mean +/- SD, p < 0.001) when treated with 108 plaque-forming units of an adenovirus containing the platelet-derived growth factor-B. Ischemic excisional wounds treated with 108 plaque-forming units of an adenovirus containing the platelet-derived growth factor-B even healed more rapidly than non-ischemic excisional wounds treated with vehicle (p < 0.05). In contrast, 5 microg of platelet-derived growth factor-BB protein (n = 2) resulted in only modest granulation tissue at the margin, but no significant differences in epithelial gap (3 +/- 0.6 mm, mean +/- SD). Plaque-forming units (106 or 108) of an adenovirus containing the beta-galactosidase transgene (n = 4) impaired wound re epithelialization with an epithelial gap of 5.11 +/- 0.69 mm, mean +/- SD, p < 0.004, and 3.8 +/- 0.57 mm, mean +/- SD, p < 0.07, respectively. Adenoviral mediated gene transfer of platelet-derived growth factor-B overcame the ischemic defect in wound healing and offers promise in the treatment of chronic nonhealing wounds. The vulnerary effects of platelet-derived growth factor-B overexpression were sufficient to overcome the adverse effects of the adenovirus or transgene on wound healing. PMID- 10469338 TI - Cell interactions control the fate of malignant keratinocytes in an organotypic model of early neoplasia. AB - The role of cell interactions during early neoplastic progression in human skin is not well understood. We report that the fate and behavior of low-grade malignant cells in stratified epithelium is dependent on their interactions with neighboring cells and with extracellular matrix during the early events in neoplastic progression. We utilized an organotypic tissue model which mimics premalignancy to monitor malignant cells (II-4) genetically marked with beta-gal and grown in the context of either normal human keratinocytes or the immortalized cell line HaCaT. HaCaT cells were permissive for clonal expansion of II-4 cells at ratios of 4:1, 12:1, and 50:1 (HAC:II-4) when compared with coculture with normal human keratinocytes. This II-4 cell expansion was associated with the failure of neighboring HaCaT cells to induce differentiation and cell cycle withdrawal of II-4, as had been seen in the context of normal human keratinocytes. When 12:1 mixtures (NHK:II-4) were stripped of all suprabasal cells and regrown, all beta-gal cells were lost showing that these normal human keratinocyte-suppressed II-4 cells had been actively sorted to a suprabasal position where their clonal expansion was limited. These growth-suppressive effects of normal human keratinocytes were found to be conditional on direct cell cell contact, as II-4 formed colonies when trypsinized from 12:1 (NHK:II-4) mixtures and grown at clonal density in submerged culture. The distribution and behavior of low-grade malignant cells was therefore dependent on the state of transformation of adjacent keratinocytes and on cell-matrix interactions. These results demonstrate that alterations in the cellular microenvironment are central to the induction of clonal expansion and early neoplastic progression in stratified epithelium. PMID- 10469339 TI - The inflammatory and cytotoxic effects of a nitric oxide releasing cream on normal skin. AB - We describe the pro-inflammatory and cytotoxic effects of nitric oxide in vivo in human skin. Nitrite and ascorbic acid were mixed on the skin of 12 normal volunteers, three times daily, to release nitric oxide. Exposure to nitric oxide was varied by randomizing the concentration of nitrite and duration of application. Nitric oxide treated skin showed significant increases in cells expressing CD3, CD4, CD8, CD68, neutrophil elastase, ICAM-1, VCAM-1, nitrosotyrosine, p53, and apoptotic cells compared with skin treated with ascorbic acid alone. There was no significant increase in mast cells. Following application of nitric oxide there were significantly fewer CD1a positive Langerhans cells in the epidermis. These appeared to lose dendritic morphology and migrate from the epidermis. There was no significant difference in staining for Ki-67, a marker related to proliferating cell nuclear antigen, between active and control skin but staining was greater after exposure to higher dose nitric oxide than the low dose. Apoptosis, cytotoxicity, and p53 staining were relatively greater after 48 h exposure than after 24 h. These results suggest that nitric oxide is pro-inflammatory and is toxic to DNA, leading to the accumulation of p53 and subsequent apoptosis. High-dose nitric oxide paradoxically led to a smaller increase in macrophages and T cells than low dose suggesting an immunosuppressive effect of higher levels. PMID- 10469340 TI - Aberrant cutaneous expression of the angiogenic factor midkine is associated with neurofibromatosis type-1. AB - Neurofibromatosis type 1 is a common autosomal dominant disorder (incidence 1:3500) characterized by lesions that include neural crest derivatives such as Schwann cells and melanocytes. A critical event in the pathogenesis of neurofibromatosis type 1 is the heterozygous germ-line loss of the tumor suppressor gene NF1. Additional genetic and/or epigenetic events have been posited, including various alterations in growth factor expression. By in situ hybridization and immunohistochemistry, we demonstrate aberrant expression of the angiogenic and tumorigenic growth factor midkine in the skin of patients with neurofibromatosis type 1, but not normal individuals. We demonstrate that midkine expression is independent of the presence of neurofibromas, and thus appears to be associated with mutations in the NF1 gene. Furthermore, midkine-containing culture media is shown to stimulate the growth of human endothelial and neurofibroma-derived cells. In conclusion, we introduce the skin as a source of dysregulated growth factors in neurofibromatosis type 1, and suggest the further study of the angiogenic factor midkine in neurofibromatosis type 1 pathogenesis. PMID- 10469341 TI - Electron diffraction provides new information on human stratum corneum lipid organization studied in relation to depth and temperature. AB - The outermost layer of mammalian skin, the stratum corneum, provides the body with a barrier against transepidermal water loss and penetration of agents from outside. The lipid-rich extracellular matrix surrounding the corneocytes in the stratum corneum is mainly responsible for this barrier function. In this study (cryo-) electron diffraction was applied to obtain information about the local lateral lipid organization in the extracellular matrix in relation to depth in human stratum corneum. For this purpose, stratum corneum grid-strips were prepared from native skin in vivo and ex vivo. It was found that the lipid packing in samples prepared at room temperature is predominantly orthorhombic. In samples prepared at 32 degrees C the presence of a hexagonal packing is more pronounced in the outer layers of the stratum corneum. Gradually increasing the specimen temperature from 30 to 40 degrees C induced a further transition from an orthorhombic to a hexagonal sublattice. At 90 degrees C all lipids were present in a fluid phase. These results are in good agreement with previously reported wide angle X-ray diffraction and Fourier transformed infrared spectroscopy studies. We conclude that the lipids in human stratum corneum are highly ordered throughout the stratum corneum and that electron diffraction allows monitoring of the local lipid organization, which contributes to the understanding of stratum corneum barrier function. PMID- 10469342 TI - Peripheral lymph node addressins are expressed on skin endothelial cells. AB - The term "peripheral node addressins" describes a set of several endothelial adhesion molecules, which collectively bind to L-selectin and react with monoclonal antibody MECA-79. They regulate lymphocyte recirculation through peripheral nodes. Their expression is thought to be restricted to a specialized vascular segment within the node, called the high endothelial venule. In certain chronic skin diseases, however, postcapillary venules of the skin may also acquire a high endothelial venule-like morphology. Employing immunohistochemistry on cryostat sections, we found these skin endothelial cells - like peripheral node high endothelial venules - to be reactive with monoclonal antibody MECA-79. Tissue lysates from the same specimens were then analyzed by immunoprecipitation using recombinant human L-selectin Fc-chimeras followed by immunoblotting using monoclonal antibody MECA-79. In contrast to peripheral node endothelium, which mainly expressed peripheral node addressin moieties of molecular sizes 90-110 kDa and 160 kDa, endothelial cells in cutaneous T cell lymphoma skin lesions expressed an additional and not yet defined 220 kDa peripheral node addressin like molecule. Most surprisingly, even in normal skin specimens, we found a distinct subset of endothelial cells located around hair follicles constitutively expressing 90-110 kDa peripheral node addressin-like moieties. It is intriguing to speculate that - in analogy to the role of peripheral node addressins in peripheral nodes - the induced expression of peripheral node addressins in chronic T cell mediated skin diseases is responsible for a sustained lymphocyte recruitment. The constitutive expression of peripheral node addressins on perifollicular endothelium may serve for a continuous lymphocyte recirculation through normal skin. PMID- 10469343 TI - Communication: expression of the novel inhibitor of apoptosis survivin in normal and neoplastic skin. AB - Apoptosis plays a fundamental part in epidermal homeostasis, and apoptotic cells have been detected in normal and diseased skin. Little is known, however, on the inhibitory mechanisms of apoptosis at the skin level. In addition to bcl-2, a novel inhibitor of apoptosis designated survivin and structurally analogous to IAP apoptosis inhibitors has been recently identified. The expression of survivin in normal and pathologic skin was investigated. Immunohistochemical studies revealed that survivin is expressed in basal keratinocytes, but not in suprabasal epidermal layers, with a pattern similar to bcl-2. In western blots, the anti survivin antibody recognized a single band of 16.5 kDa in protein extracts from normal human keratinocytes in culture, in agreement with the predicted size of survivin. In addition, survivin immunoreactivity was detected in benign and malignant melanocytic lesions, with strong expression in invasive lesions of melanomas. Whereas survivin staining was undetectable in benign epithelial tumors, such as seborrheic keratoses, it was observed in all epidermal layers in Bowen's disease. Interestingly, at variance with bcl-2, survivin was markedly expressed in squamous cell carcinoma, but virtually lacking in basal cell carcinoma, suggesting that these two apoptosis inhibitors may act through different anti-apoptotic pathways. Deregulation of survivin may influence both epidermal homeostasis and the development of melanoma and nonmelanoma skin cancer. PMID- 10469344 TI - Compound heterozygosity for silent and dominant glycine substitution mutations in COL7A1 leads to a marked transient intracytoplasmic retention of procollagen VII and a moderately severe dystrophic epidermolysis bullosa phenotype. PMID- 10469345 TI - Plectin serves as an autoantigen in paraneoplastic pemphigus. PMID- 10469346 TI - Correlation between antioxidants and phototypes in melanocytes cultures. A possible link of physiologic and pathologic relevance. PMID- 10469347 TI - Retardation of hair follicle development by the deletion of TrkC, high-affinity neurotrophin-3 receptor. PMID- 10469348 TI - Animal models of psoriasis. PMID- 10469349 TI - Chronic allograft nephropathy: An update. AB - Chronic allograft nephropathy is the most prevalent cause of renal transplant failure in the first post-transplant decade, but its pathogenesis has remained elusive. Clinically, it is characterized by a slow but variable loss of function, often in combination with proteinuria and hypertension. The histopathology is also not specific, but transplant glomerulopathy and multilayering of the peritubular capillaries are highly characteristic. Several risk factors have been identified, such as advanced donor age, delayed graft function, repeated acute rejection episodes, vascular rejection episodes, and rejections that occur late after transplantation. A common feature of chronic allograft nephropathy is that it develops in grafts that have undergone previous damage, although the mechanism(s) responsible for the progressive fibrosis and tissue remodeling has not yet been defined. Hypotheses to explain chronic allograft nephropathy include the immunolymphatic theory, the cytokine excess theory, the loss of supporting architecture theory, and the premature senescence theory. The most effective option to prevent chronic allograft nephropathy is to avoid graft injury from both immune and nonimmune mechanisms. PMID- 10469350 TI - Role of vascular endothelial growth factor in the regulation of angiogenesis. AB - Compelling evidence indicates that vascular endothelial growth factor (VEGF) is a fundamental regulator of normal and abnormal angiogenesis. The loss of a single VEGF allele results in defective vascularization and early embryonic lethality. VEGF plays also a critical role in kidney development, and its inactivation during early postnatal life results in the suppression of glomerular development and kidney failure. Recent evidence indicates that VEGF is also essential for angiogenesis in the female reproductive tract and for morphogenesis of the epiphyseal growth plate and endochondral bone formation. Substantial experimental evidence also implicates VEGF in pathological angiogenesis. Anti-VEGF monoclonal antibodies or other VEGF inhibitors block the growth of several human tumor cell lines in nude mice. Furthermore, the concentrations of VEGF are elevated in the aqueous and vitreous humors of patients with proliferative retinopathies such as the diabetic retinopathy. In addition, VEGF-induced angiogenesis results in a therapeutic benefit in several animal models of myocardial or limb ischemia. Currently, both therapeutic angiogenesis using recombinant VEGF or VEGF gene transfer and inhibition of VEGF-mediated pathological angiogenesis are being pursued clinically. PMID- 10469351 TI - Syntaxin 3 and Munc-18-2 in epithelial cells during kidney development. AB - BACKGROUND: Differentiation of epithelial cells involves the assembly of polarized membrane transport machineries necessary for the generation and maintenance of the apical and basolateral membrane domains characteristic of this cell type. We have analyzed the expression patterns of vesicle-docking proteins of the syntaxin family in mouse kidney, focusing on syntaxin 3 and its interaction partner, the Sec1-related Munc-18-2. METHODS: Expression patterns were studied by in situ hybridization and immunocytochemistry and the complex formation of syntaxin 3 and Munc-18-2 by coimmunoprecipitation and Western blotting. RESULTS: We have previously shown by in situ hybridization that Munc-18 2 is present in the proximal tubules and collecting ducts of embryonic day 17 mouse kidney. We compared this with the expression patterns of syntaxin 1A, 2, 3, 4, and 5, and found that syntaxin 3 was enriched in the same epithelial structures in which Munc-18-2 was abundant. By immunocytochemistry, the two proteins colocalized at the apical plasma membrane of proximal tubule and collecting duct epithelial cells, and they were shown to form a physical complex in the kidney. The expression of both proteins was up-regulated during kidney development. The most prominent changes in expression levels coincided with the differentiation of proximal tubules, suggesting a role in the generation of the highly active reabsorption machinery characterizing this segment of the nephron. CONCLUSION: The results show that Munc-18-2 and syntaxin 3 form a complex in vivo and suggest that they participate in epithelial cell differentiation and targeted vesicle transport processes in the developing kidney. PMID- 10469352 TI - cDNA cloning and embryonic expression of mouse nuclear pore membrane glycoprotein 210 mRNA. AB - BACKGROUND: In embryonic kidneys, mesenchymal cells convert into epithelium in response to an induction by the tip of the ureter bud. Metanephric mesenchyme can also be induced to convert into epithelium in vitro. It is a model system to identify genes that could be important for epithelial development. METHODS: By differential screening of a cDNA library made from mesenchymes induced in transfilter cultures by embryonic spinal cord for 24 hours, we selected cDNA clones representing genes that were preferentially expressed in 24-hour-induced mesenchyme and not in uninduced mesenchyme. The sequence of one clone was determined and used to obtain the sequence of a complete open reading frame. By Northern blotting and in situ hybridization, the expression of the mRNA in embryonic kidneys was determined. RESULTS: We report the sequence and expression pattern of a marker for the 24-hour-induced state, mouse nuclear pore membrane glycoprotein 210 (mPOM210). The deduced 1886 amino acid sequence shows a 95% identity to the sequence of rat gp210. Northern blotting revealed a single 7.5 kb mRNA in 24-hour-induced mesenchyme, whereas message levels were fourfold to fivefold lower in uninduced mesenchyme. In situ hybridization of in vivo development confirmed the preferential expression of mPOM210 in epithelial cells. In the kidney, expression was seen in both the epithelium derived from the ureteric tree and the mesenchyme-derived epithelium. In other tissues of 13-day old embryos, expression was also confined to the epithelium. In nervous tissues, the olfactory epithelium and walls of the lateral ventricle were the most prominently stained. Weak expression was seen in the heart. CONCLUSIONS: mPOM210 mRNA is an early marker for developing epithelial cells. Furthermore, our results suggest that nuclear pore membrane proteins could be more cell-type specific than previously anticipated. PMID- 10469353 TI - Modulation of interleukin-6 by beta2-adrenoceptor in endotoxin-stimulated renal macrophage cells. AB - BACKGROUND: Activation of the cAMP signaling pathway by means of beta2 adrenoceptor agonists has been shown to up-regulate interleukin-6 (IL-6) gene expression and to stimulate IL-6 production in macrophage cells. However, whether beta2-adrenoceptor activation can also modify the rate of IL-6 production in macrophage cells activated by the bacterial endotoxins has not yet been determined. Using renal resident macrophage cells treated with endotoxin, lipopolysaccharide (LPS), and beta2-adrenoceptor agonist, terbutaline, we investigated the role of cAMP pathway, tumor necrosis factor (TNF)-alpha and mitogen-activated protein kinase (MAPK) pathway (p42/p44) in regulating IL-6 production. METHODS: IL-6 protein, mRNA, and promoter activity were measured in these cells exposed to LPS (1 microg/ml) and/or terbutaline (10(-9) to 10(-6) M). Furthermore, the time course effects of terbutaline on cAMP, MAPK (p42/p44), and TNF-alpha release were evaluated in the cells. RESULTS: Terbutaline at high concentrations (10(-6) M) significantly up-regulated IL-6 by approximately 25% (P<0.05), whereas at a lower concentration (10(-8) M), it down-regulated IL-6 production by 42% (P<0.05). Terbutaline (10(-8) and 10(-6) M) caused a concentration- and time-dependent stimulation of cAMP (P<0.05) and TNF production (P<0.05) and a time-dependent decrease in MAPK activity (P<0.05). Following the addition of a cAMP inhibitor, IL-6 promoter activity was correlated with TNF alpha levels and MAPK activity. CONCLUSIONS: A biphasic effect of beta2 adrenoceptor agonist on IL-6 production in renal resident macrophage cells became apparent when LPS was exposed to the cells. The terbutaline-induced down regulation of IL-6 gene production was mediated by an inhibitory effect of terbutaline on TNF-alpha, which was exerted through the MAPK and cAMP pathways, whereas the up-regulation appeared to be due to a direct action of intracellular cAMP. PMID- 10469354 TI - Aminopeptidase A activity and angiotensin III effects on [Ca2+]i along the rat nephron. AB - BACKGROUND: This study examined the specific effects of angiotensin III (Ang III) along the nephron. METHODS: We examined the distribution of aminopeptidase A (APA) activity by using a specific APA inhibitor and by immunostaining with an antirat kidney APA antibody, the Ang III-induced variations of [Ca2+]i by using fura-2 and the characterization of the receptor subtype involved in the response to Ang III in cortical thick ascending limb (CTAL). RESULTS: APA activity was found all along the nephron but was higher in the cortex than in the medulla. This was confirmed by immunostaining. Increases in [Ca2+]i elicited by 10(-7) mol/liter Ang III were observed all along the nephron. The characterization of the receptor subtype involved in the [Ca2+]i response to Ang III in CTAL indicated that EC50 values for Ang III and Ang II were similar (13.5 and 10.3 nmol/liter, respectively), and Ang III-induced responses were totally abolished by AT1 receptor but not by AT2 receptor antagonists. There was a cross desensitization of [Ca2+]i responses to 10(-7) mol/liter Ang III and Ang II, and the [Ca2+]i responses to 10(-7) mol/liter Ang II and Ang III were not additive. CONCLUSION: These results show that in CTAL, the [Ca2+]i responses to Ang II and Ang III occur through the same AT1a receptor because this subtype is predominant in this segment. Taken together, these data suggest that APA could be a key enzyme to generate Ang III from Ang II in the kidney. PMID- 10469355 TI - Leptin stimulates proliferation and TGF-beta expression in renal glomerular endothelial cells: potential role in glomerulosclerosis [seecomments]. AB - BACKGROUND: Leptin inhibits food intake and increases energy expenditure. Although the kidney expresses abundant transcripts of the short form of the leptin receptor (Ob-Ra), a role for this hormone in renal function remains unclear. Because individuals with massive obesity who may exhibit increased leptin serum concentrations develop renal glomerulosclerosis, we studied whether leptin can influence renal growth and profibrogenic processes. METHODS: The effects of recombinant leptin on proliferation and synthesis of transforming growth factor-beta1 (TGF-beta1) was investigated in cultured glomerular endothelial cells of the rat (GERs) and syngeneic mesangial cells. Furthermore, leptin receptor expression and potential signal transduction pathways were evaluated in GERs. In addition, leptin was also infused for different time periods (72 hr and 3 weeks) into naive rats. RESULTS: Recombinant mouse leptin induced proliferation of GERs, but not of syngeneic mesangial cells. Coincubation with angiotensin II and leptin exerts additive proliferative effects in GERs. An antileptin-receptor antibody totally abolished this proliferation but did not influence serum-induced proliferation. GER expressed high affinity receptors of the Ob-Ra type (Kd, 4 nM; Bmax, 9700 receptors/cell). Leptin also stimulated phosphorylation of STAT1alpha, and kinase inhibitors attenuated proliferation, suggesting a pivotal role of phosphorylation in this process. Incubation of GERs with leptin also induced mRNA expression of TGF-beta1 and enhanced secretion of this profibrogenic cytokine. Short-term leptin infusion (72 hr) into naive rats induced a significant proliferation, mainly restricted to glomerular endothelial cells, and enhanced glomerular TGF-beta1 mRNA levels. In rats continuously infused for three weeks with leptin, glomerular TGF-beta1 expression was still enhanced, and an additional increase in glomerular collagen type IV mRNA and protein expression was detected. These animals revealed an increase in proteinuria compared with control-infused rats. CONCLUSION: Our findings are the first in vitro and in vivo demonstration that leptin is a renal growth and profibrogenic factor. These results may be an important contribution to our understanding of how leptin can contribute to renal damage, characterized by endocapillary proliferation and subsequent development of glomerulosclerosis, in pathophysiological situations with high circulating levels such as in diabetics or obese individuals. Although the effects of leptin itself are moderate, growth promoting and profibrogenic effects may be enhanced in concert with other factors such as angiotensin II. PMID- 10469356 TI - c-Rel and p65 trans-activate the monocyte chemoattractant protein-1 gene in interleukin-1 stimulated mesangial cells. AB - BACKGROUND: The chemokine monocyte chemoattractant protein-1 (MCP-1) is secreted by human glomerular mesangial cells in response to interleukin-1 (IL-1) and has a central role in amplifying the inflammatory response during glomerulonephritis. However, the mechanism by which IL-1 regulates its transcription is not understood. Specific members of the nuclear factor kappaB/rel (NF-kappaB) proteins may regulate MCP-1 expression in a stimulus- and tissue-specific manner. METHODS: Electrophoretic mobility shift assays and Western blot analysis characterized the members of the NF-kappaB family that bound the two NF-kappaB sites of the MCP-1 enhancer (A1 and A2) in vitro. Trans-activation of the MCP-1 gene was investigated by transfer of the MCP-1 enhancer DNA to mesangial cells. RESULTS: Primary human mesangial cells contained in addition to p50 (NF-kappaB1) and p65 (Rel A) NF-kappaB proteins, the oncoprotein c-rel, and Rel B, but not p52 (NF-kappaB2). IL-1 induced c-rel to form a complex with p65, which bound the MCP 1 A2 site but not the A1 or IL-6 NF-kappaB sites in vitro. IL-1 up-regulated transfected MCP-1 enhancer activity. Cotransfer of the MCP-1 enhancer together with individual members of the NF-kappaB family showed that the heterodimer c relp65 or (p65)2 can selectively trans-activate the MCP-1 gene via its A1 and A2 sites in mesangial cells. CONCLUSIONS: This study demonstrates for the first time that the c-rel oncoprotein can enhance MCP-1 transcription in mesangial cells and suggests that it may have an important role in amplifying gene expression in the inflamed glomerulus. PMID- 10469357 TI - Localization of fibroblast growth factor-2 (basic FGF) and FGF receptor-1 in adult human kidney. AB - BACKGROUND: The expression pattern of fibroblast growth factor-2 (FGF-2; basic FGF), a pleiotrophic growth factor, as well as one of its receptors (FGFR1), in the kidney is highly controversial. METHODS: Using an approach that combines multiple antibodies for immunohistochemistry and correlative in situ hybridization, we assessed the intrarenal expression of both FGF-2 and FGFR1 in 13 specimens of adult kidney removed during tumor nephrectomy. RESULTS: The FGF-2 expression pattern in the kidneys as detected by immunohistochemistry was variable and depended on the antibody used. The most consistent expression of FGF 2 protein was demonstrated in glomerular parietal epithelial cells, tubular cells (mainly of the distal nephron), as well as arterial endothelial cells. These locations also corresponded to areas of FGF-2 mRNA expression. Additionally, by immunohistochemistry, FGF-2 protein was detected in arterial smooth muscle cells and occasional podocytes. The expression of FGFR1 protein and mRNA was most consistently present in tubular cells of the distal nephron and in vascular smooth muscle cells. In situ hybridization, but not immunohistochemistry, also suggested FGFR1 expression in cells that could not be precisely identified within the glomerular tuft as well as some interstitial cells. CONCLUSION: These data suggest potential autocrine and paracrine pathways within the FGF-2 system, particularly within the vascular walls and in the distal nephron, and thereby provide information for further mechanistic understanding of the role of the FGF 2 system in human renal disease. PMID- 10469358 TI - Effect of candesartan cilexetil (TCV-116) in rats with chronic renal failure. AB - BACKGROUND: Inhibition of the renin-angiotensin system by both angiotensin II type 1 receptor antagonists (AT1As) and angiotensin I-converting enzyme inhibitors (ACEIs) shows renoprotective effects in rats with chronic renal failure when treatment is started in the early phase of renal injury. In this study, we examined the renal protective effects of candesartan cilexetil (TCV 116), an AT1A, and enalapril, an ACEI, in the progressive phase of renal injury in 5/6 nephrectomized rats. METHODS: Candesartan cilexetil (1 mg/kg/day) and enalapril (10 mg/kg/day) were orally administered once a day for 4 weeks (the short-term experiment) or 16 weeks (the long-term experiment) to 5/6 nephrectomized rats beginning 15 weeks after the nephrectomy, that is, after they had already showed marked proteinuria. RESULTS: In vehicle-treated rats, proteinuria, glomerulosclerosis, and interstitial fibrosis developed. Moreover, enhanced expression of transforming growth factor-beta1 (TGF-beta1) in the injured glomeruli was observed. These adverse changes progressed with time, and in the short-term experiment, both drugs inhibited them. In the long-term experiment, the progressive proteinuria and the elevation of blood pressure were similarly attenuated by both drugs. However, candesartan cilexetil significantly inhibited the progression of glomerulosclerosis, the expression of TGF-beta1, and interstitial fibrosis, whereas enalapril did not. CONCLUSION: These results indicate that candesartan cilexetil shows potent and long-term preventive effects against the progression of previously developed renal injury. PMID- 10469359 TI - Dynamics of E-cadherin and gamma-catenin complexes during dedifferentiation of polarized MDCK cells. AB - BACKGROUND: E-cadherin mediated cell-cell adhesion and hepatocyte growth factor (HGF) are important for renal epithelial morphogenesis. We previously showed that HGF dedifferentiates previously well polarized Madin-Darby canine kidney (MDCK) cell monolayers grown on filters. The regulation of E-cadherin during epithelial dedifferentiation is not known. We hypothesized that E-cadherin mediated cell cell adhesion is modulated during HGF induced dedifferentiation of MDCK cell monolayers. METHODS: We analyzed E-cadherin/gamma-catenin interaction and distribution during epithelial dedifferentiation in vitro using a model of polarized MDCK cell monolayers treated with HGF. RESULTS: Surface immunoprecipitation experiments showed that HGF increased the amount of cell surface E-cadherin associated with gamma-catenin. Biochemical and morphological examination of the TX-100 solubility of junctional E-cadherin and gamma-catenin in control and HGF treated cells showed an increase in solubility of only E cadherin during loss of cell polarity. Metabolic labeling of control and HGF treated cells showed that HGF stimulated the synthetic rate of E-cadherin and gamma-catenin molecules. Inulin flux across MDCK cell monolayers increases with HGF treatment. CONCLUSION: These data provide evidence for both the dissociation of E-cadherin molecules from the actin cytoskeleton and an increase in the total number of E-cadherin/gamma-catenin complexes on the cell surface during HGF induced dedifferentiation of polarized renal epithelium. These data support the hypothesis that E-cadherin function is inhibited by a mechanism of detachment from the actin based cytoskeleton during HGF induced dedifferentiation of polarized renal epithelia. PMID- 10469360 TI - Protection from toxicant-mediated renal injury in the rat with anti-CD54 antibody. AB - BACKGROUND: The benefit of the potent chemotherapeutic agent cisplatin in treating neoplasms is limited by nephrotoxicity. We tested the hypothesis that CD54 [intercellular adhesion molecule-1 (ICAM-1)] is an important mediator in cisplatin-mediated renal failure. METHODS: The effect of a monoclonal anti-CD54 antibody was evaluated in a rat model of cisplatin toxicity. Renal function, histopathology, renal myeloperoxidase activity, and mortality were determined in the anti-CD54 and placebo groups. RESULTS: Renal CD54 mRNA expression was markedly increased by 24 hours after exposure to cisplatin in mice. An improvement in renal function, mortality, and histological abnormalities was evident in animals exposed to cisplatin and treated with anti-CD54 antibody (mAb). Seven days after the administration of cisplatin, the mean creatinine was 0.65+/-0.05 mg/dl in the rats that received anti-CD54 mAb and 4.76+/-1.42 in control animals (P<0.02). Mortality was lower in experimental animals (0 vs. 29% in control rats seven days following cisplatin, P<0.04). Histological evidence of cell injury was markedly attenuated (P<0.04) in the treated compared with the control rats. CONCLUSION: CD54 may be critical in the pathophysiology of renal injury following cisplatin, perhaps by its effects on leukocyte-endothelial interactions. PMID- 10469361 TI - Pentoxifylline attenuates experimental mesangial proliferative glomerulonephritis. AB - BACKGROUND: Accumulation of glomerular macrophages, proliferation of mesangial cells (MCs), and deposition of extracellular matrix proteins are pathobiological hallmarks of glomerulonephritis. We previously reported that a clinically available nonselective inhibitor of cyclic 3',5'-nucleotide phosphodiesterase, pentoxifylline (PTX), inhibits proliferation of cultured rat MCs, as well as collagen production by these cells. In this study, we investigated the in vivo effects of PTX on rat anti-Thy1 disease, a model of mesangial proliferative nephritis. METHODS: Anti-Thy1 nephritis was induced in Sprague-Dawley rats by injecting mouse anti-rat Thy1 antibodies intravenously. Nephritic rats were randomly assigned to receive PTX (0.1 g/kg/day) or vehicle (phosphate-buffered saline) and were sacrificed at various time points. Paraffin kidney sections were stained with hematoxylin and periodic acid-Schiff reagents for glomerular histology. Frozen kidney sections were stained by monoclonal antibodies against proliferating cell nuclear antigen, ED-1, and alpha-smooth muscle actin and were visualized by color development from a horseradish peroxidase reaction. Monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1), and various extracellular matrix mRNAs were analyzed by Northern blotting. Urine protein concentrations were determined by Lowry's method. RESULTS: Nephritic rats treated with PTX excreted less urinary protein on day 5 of nephritis than vehicle treated nephritic rats. In periodic acid-Schiff-stained kidneys from PTX-treated nephritic rats, there was attenuation of both glomerular cellularity and glomerular sclerosis compared with vehicle-treated nephritic rats. PTX decreased the augmented glomerular mRNA levels of MCP-1 and ICAM-1 at two hours and on day 1 of nephritis. Immunoreactive staining showed that PTX reduced the number of proliferating glomerular macrophages on days 1, 2, and 3, but not at two hours of nephritis, compared with vehicle-treated nephritic rats. On day 5, PTX decreased the number of activated proliferating MCs and attenuated the glomerular mRNA levels of type I (alpha1), type III (alpha1), and type IV (alpha1) collagen and fibronectin compared with vehicle-treated nephritic rats. CONCLUSION: The administration of PTX to rats with anti-Thy1 disease reduces accumulation and proliferation of glomerular macrophages, attenuates proteinuria, suppresses activation and proliferation of MCs, and ameliorates glomerular sclerosis. These results suggest that PTX may have a suppressive effect in acute phases or relapses of mesangial proliferative glomerulonephritis. PMID- 10469362 TI - Gender differences in renal growth and function after uninephrectomy in adult rats. AB - BACKGROUND: It is known that compensatory renal growth (CRG) following unilateral nephrectomy (UNX) increases both the size of the kidney and its functional capacity; however, few studies have investigated whether differences in CRG exist between the sexes. Our study examined the sex-related differences in remnant kidney growth and function two months following UNX. METHODS: Adult male and female Wistar rats underwent either left UNX or sham operation and recovered for 8 to 10 weeks. Another group of female rats underwent ovariectomy (Ox), with vehicle, estrogen, or testosterone replacement: two-weeks postsurgery animals underwent UNX and recovered for 8 to 10 weeks. Metabolic studies, acute renal function studies [response to acute saline volume expansion (2 to 4% of body wt) or phosphate (Pi) infusions in thyroparathyroidectomized rats (to determine the transport maximum (TmPi)], and renal morphology were assessed at the end of the experimental period. RESULTS: Two months post-UNX, male remnant kidneys grew 114+/-7% of their excised kidney weight (KW), whereas female remnant kidneys grew only 57+/-4% (P<0.05). There was a significant increase in the glomerular volume of male remnant kidneys (126.2+/-13.4%, P<0.001) compared with control kidney volume, whereas there was no change in glomerular volume in female remnant kidneys (20.2+/-16.1%, P = NS). There was also glomerular and tubular damage in the male remnant kidneys, whereas female remnant kidneys were intact. Studies in Ox female rats supplemented with gonadal steroids determined that testosterone is the driving force for the enhanced remnant kidney growth and glomerular hypertrophy. Renal function studies determined that UNX males had significantly higher glomerular filtration rates (GFRs) than UNX females, although the GFR/single KW was not different between the sexes, indicating a proportional increase in GFR. Basal urinary sodium excretion and urine flow rates were significantly higher in anesthetized UNX rats than their sham-operated controls, and urinary sodium excretion and urine flow rates in UNX males were significantly higher than in UNX females. Both male and female UNX rats responded to volume expansion with an exaggerated initial sodium and urine excretion compared with their controls. Phosphate handling was not altered in UNX male rats; however, UNX female rats had increases in fractional Pi excretion that were associated with significant reductions in the maximum capacity for Pi reabsorption (2.10+/-0.07 vs. 3.43+/-0.24 micromol/ml GFR in female controls, P<0.0001). This difference was also observed in Ox rats treated with estrogen and testosterone (2.31+/-0.07 vs. 3.12+/-0.11 micromol/ml GFR, P<0.0007). CONCLUSIONS: These findings indicate that sexual dimorphism exists in remnant kidney growth and function two months following UNX. Indeed, morphological abnormalities and impairment in renal phosphate handling are affected by gonadal steroids by two-months post-UNX. The fact that renal Pi transport was reduced in female but not male UNX rats may also have important implications during periods of high metabolic demand for phosphate in the female. PMID- 10469363 TI - Inhibition of glomerular cell apoptosis by heparin. AB - BACKGROUND: Heparin, the multifunctional glycosaminoglycan, has been considered a therapeutic agent for glomerular diseases. Although a number of biological properties are postulated to explain its therapeutic utility, it is unknown whether heparin affects cell survival in the glomerulus. In this report, we investigated the effect of heparin on apoptosis of glomerular cells. METHODS: Cultured rat mesangial cells were pretreated with heparin or heparan sulfate proteoglycan (HSPG) and were exposed to proapoptotic stimuli. To examine an effect of heparin on spontaneous apoptosis that occurs in explanted glomeruli, isolated rat glomeruli were incubated in the presence or absence of heparin. Apoptosis was evaluated by Hoechst 33258 staining, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling, and agarose gel electrophoresis to detect DNA fragmentation. The effect of heparin on activator protein 1 (AP-1), a crucial mediator for oxidant-induced apoptosis, was examined by Northern blot analysis and a reporter assay. RESULTS: Heparin and HSPG inhibited apoptosis of mesangial cells triggered by hydrogen peroxide. It was associated with blunted expression of c-fos/c-jun mRNAs and suppression of AP-1 activation. The cytoprotective effect of heparin was also observed in other cell types and in apoptosis triggered by different stimuli. That is, (a) heparin inhibited mesangial cell apoptosis induced by staurosporine, pyrrolidine dithiocarbamate, and ultraviolet light, and (b) heparin suppressed oxidant-induced apoptosis of NRK49F fibroblasts and Madin-Darby canine kidney epithelial cells. Furthermore, heparin attenuated spontaneous apoptosis of podocytes in explanted glomeruli. CONCLUSIONS: These results indicate the novel potential of heparin as an inhibitor of apoptosis in several cell types, including glomerular cells. PMID- 10469364 TI - Dissociation of mesangial cell migration and proliferation in experimental glomerulonephritis. AB - BACKGROUND: Recently, we documented that following in vivo mesangial cell (MC) ablation in the Thy1 model, reconstitution of the mesangium occurs by a coordinated proliferation and migration of Thy1 (OX-7)-positive cells originating from the hilus and extraglomerular mesangium. We investigated the role of basic fibroblast growth factor (bFGF) in the mediation of these events. METHODS: Rats were injected with antithymocyte serum and 48 hours later were pulsed with 3H thymidine to label proliferating cells. Ninety minutes later, a baseline renal biopsy was obtained, and rats were injected with neutralizing anti-bFGF antibodies or control IgG. Sacrificial biopsies were obtained at 96 hours of disease. Using computer image analysis, biopsies from both time points were quantitated for the number of radiolabeled MC (proliferation) and their mean distance from the hilus (migration). The effect of bFGF on the migration of MCs in culture was examined using a chemotactic assay. RESULTS: At sacrifice, autoradiographs of rats receiving anti-bFGF had significantly fewer radiolabeled MCs as compared with rats receiving control IgG (8.7+/-1.9 vs. 14.7+/-3.5, P = 0.0001), yielding an overall 40% reduction in proliferation. There was no difference, however, in the final distance of radiolabeled MCs from the glomerular hilus in the two groups, indicating that the administration of anti bFGF did not effect MC migration in this model. In an in vitro chemotactic assay, MCs migrated in response to platelet-derived growth factor (PDGF) BB (20 ng/ml), but did not migrate in response to bFGF at a wide range of concentrations (0.5 to 50 ng/ml). CONCLUSIONS: These studies demonstrate that bFGF is an important mediator of MC proliferation but that it does not significantly influence MC migration. This is the first demonstration showing that the mediators effecting proliferation can be dissociated from those mediating migration in renal injury. PMID- 10469365 TI - Inhibitors of poly (ADP-ribose) synthetase protect rat proximal tubular cells against oxidant stress. AB - BACKGROUND: The generation of reactive oxygen species (ROS) has been implicated in the pathogenesis of renal ischemia-reperfusion injury. ROS produce DNA strand breaks that lead to the activation of the DNA-repair enzyme poly (ADP-ribose) synthetase (PARS). Excessive PARS activation results in the depletion of its substrate, nicotinamide adenine dinucleotide (NAD) and subsequently of adenosine 5'-triphosphate (ATP), leading to cellular dysfunction and eventual cell death. The aim of this study was to investigate the effect of various PARS inhibitors on the cellular injury and death of rat renal proximal tubular (PT) cells exposed to hydrogen peroxide (H2O2). METHODS: Rat PT cell cultures were incubated with H2O2 (1 mM) either in the presence or absence of the PARS inhibitors 3-aminobenzamide (3-AB, 3 mM), 1,5-dihydroxyisoquinoline (0.3 mM) or nicotinamide (Nic, 3 mM), or increasing concentrations of desferrioxamine (0.03 to 3 mM) or catalase (0.03 to 3 U/ml). Cellular injury and death were determined using the MTT and lactate dehydrogenase (LDH) assays, respectively. H2O2-mediated PARS activation in rat PT cells and the effects of PARS inhibitors on PARS activity were determined by measurement of the incorporation of [3H]NAD into nuclear proteins. RESULTS: Incubation of rat PT cells with H2O2 significantly inhibited mitochondrial respiration and increased LDH release, respectively. Both desferrioxamine and catalase reduced H2O2-mediated cellular injury and death. All three PARS inhibitors significantly attenuated the H2O2-mediated decrease in mitochondrial respiration and the increase in LDH release. Incubation with H2O2 produced a significant increase in PARS activity that was significantly reduced by all PARS inhibitors. 3-Aminobenzoic acid (3 mM) and nicotinic acid (3 mM), structural analogs of 3-AB and Nic, respectively, which did not inhibit PARS activity, did not reduce the H2O2-mediated injury and necrosis in cultures of rat PT cells. CONCLUSION: We propose that PARS activation contributes to ROS-mediated injury of rat PT cells and, therefore, to the cellular injury and cell death associated with conditions of oxidant stress in the kidney. PMID- 10469366 TI - TGF-beta1 down-regulates induced expression of both class II MHC and B7-1 on primary murine renal tubular epithelial cells. AB - BACKGROUND: We examined the immunomodulatory effects of transforming growth factor-beta1 (TGF-beta1) on the regulation of class II MHC and costimulatory molecule expression in a primary renal tubular epithelial cell line, called F1K. METHODS: Class II major histocompatibility complex (MHC), class II transactivator, B7-1, intercellular adhesion molecule-1 (ICAM-1) and interferon gamma (IFN-gamma) receptor beta chain were evaluated in untreated and cytokine treated F1K by Northern hybridization analysis and flow cytometry. T cell activation studies were performed to assess TGF-beta1-mediated effects on antigen presenting cell function of F1K. RESULTS: Pretreatment of F1K with TGF-beta1 markedly inhibited IFN-gamma-induced class II MHC expression, by both FACS and Northern analysis. Total class II transactivator mRNA levels were also diminished by TGF-beta1, indicating that class II MHC modulation in F1K results from inhibition of this intermediate protein. As previous studies demonstrated that cotreatment of F1K cells with IFN-gamma + lipopolysaccharide (LPS) induces B7-1, we evaluated the potential regulatory effects of TGF-beta1 exposure on B7-1 expression. Our studies revealed that B7-1 mRNA and cell-surface expression in IFN-gamma + LPS-treated F1K were decreased by TGF-beta1 pretreatment. Functional studies evaluating TGF-beta1-mediated effects were performed with IFN-gamma + LPS treated F1K and MR1.3, a nephritogenic CD4+ Th2 clone derived from kidneys of animals with autoimmune glomerulonephritis. Interleukin (IL)-4 production assays demonstrated activation of MR1. 3 by IFN-gamma + LPS-treated cells, but not by IFN-gamma + LPS-treated cells previously exposed to TGF-beta1, indicating that TGF-beta1-mediated inhibition of class II MHC and B7-1 expression alters the antigen presenting cell function of F1K. CONCLUSIONS: These studies describe the proscriptive influence of TGF-beta1 on class II MHC and B7-1 expression in renal tubular epithelial cells. Such findings indicate that TGF-beta1 alters the antigen presenting cell function of renal tubular epithelial cells in vitro, and suggest a potential mechanism for immunosuppression of T cell-mediated renal immune responses in vivo. PMID- 10469368 TI - Increased oxidative stress in mouse kidneys with unilateral ureteral obstruction. AB - BACKGROUND: Unilateral ureteral obstruction (UUO) is a well-established experimental model of renal injury leading to interstitial fibrosis. The molecular and cellular mechanism(s) of interstitial fibrosis in UUO kidney is beginning to be elucidated. Oxidative stress has been implicated in the pathogenesis of various forms of renal injury; however, little is known about its involvement in the setting of ureteral obstruction. METHODS: To investigate the possible involvement of oxidative stress in the obstructive nephropathy, we studied the occurrence and distribution of Nepsilon-carboxymethyl-lysine (CML) in the kidneys after ureteral obstruction. CML is an integrative biomarker of the cumulative protein damage induced by glycoxidation. Heme oxygenase-1 (HO-1) mRNA and protein expression, which is a sensitive and reliable indicator of oxidative stress, were also examined. RESULTS: CML immunoreactivity was found in the interstitium of UUO kidneys 10 days after the onset ureteral obstruction. HO-1 mRNA was up-regulated as early as 12 hours after ureteral obstruction. HO-1 immunoreactivity was observed in the periglomerular and peritubular interstitium two days after ureteral obstruction. CONCLUSIONS: These results strongly suggested the presence of increased oxidative stress in the interstitium of UUO kidneys. The oxidative stress and the formation of various kind of biological active oxidative products in the interstitium are supposed to play significant roles in UUO kidney. PMID- 10469367 TI - MIP-1alpha and MCP-1 contribute to crescents and interstitial lesions in human crescentic glomerulonephritis. AB - BACKGROUND: The precise molecular mechanisms of macrophage (Mphi) recruitment and activation in crescentic glomerulonephritis remain to be investigated. We hypothesized that locally produced macrophage inflammatory protein (MIP)-1alpha and monocyte chemoattractant protein (MCP)-1 via the chemokine receptors participate in the pathophysiology of human crescentic glomerulonephritis by recruiting and activating Mphi. METHODS: We investigated the levels of MIP-1alpha and MCP-1 by enzyme-linked immunosorbent assay (ELISA) in 20 healthy subjects, 20 patients with crescentic glomerulonephritis, and 41 control patients with various other renal diseases. The presence of MIP-1alpha, MCP-1, and the cognate chemokine receptor for MIP-1alpha, CCR5, in the diseased kidneys was evaluated by immunohistochemical and in situ hybridization analyses. RESULTS: MIP-1alpha positive cells were mainly detected in crescentic lesions, whereas MCP-1 was mainly in the interstitium. In addition, we detected CCR5-positive cells in diseased glomeruli and interstitium. Urinary MIP-1alpha was detected in crescentic glomerulonephritis, even though it was below detectable levels in healthy subjects and in patients with other renal diseases without crescents. Urinary MIP-1alpha levels in the patients with crescentic glomerulonephritis were well correlated with the percentage of cellular crescents and the number of CD68 positive infiltrating cells and CCR5-positive cells in the glomeruli. However, urinary MCP-1 levels were well correlated with the percentage of both total crescents and fibrocellular/fibrous crescents and the number of CD68-positive infiltrating cells in the interstitium. Moreover, elevated urinary levels of both MIP-1alpha and MCP-1 dramatically decreased during glucocorticoid therapy-induced convalescence. CONCLUSIONS: These observations suggest that locally produced MIP 1alpha may be involved in the development of cellular crescents in the acute phase via CCR5 and that MCP-1 may be involved mainly in the development of interstitial lesions in the chronic phase when fibrocellular/fibrous crescents are present, possibly through Mphi recruitment and activation. PMID- 10469369 TI - Mechanism of FK 506/520 action on rat renal proximal tubular Na+, K+-ATPase activity. AB - BACKGROUND: The neurotransmitter in renal sympathetic nerves, norepinephrine (NE), regulates the activity of proximal tubule (PT) Na+,K+-ATPase in a bidirectional manner via stimulation of alpha- and beta-adrenoceptors. The stimulatory alpha-adrenergic pathway is mediated by calcineurin, the target molecule for FK 506 and related compounds. We examined whether the FK 506 analogue FK 520, by interrupting the calcineurin-mediated alpha-adrenergic signaling pathway, enhance the inhibitory beta-adrenergic effect of NE on PT Na+,K+-ATPase activity. METHODS: The effects of three days of treatment with FK 520 were examined on rat renal PT Na+,K+-ATPase activity, measured as ouabain sensitive ATP hydrolysis in single, microdissected PT segments. Renal function studies, including glomerular filtration rate (GFR) and urinary excretion of N acetyl-3-D-glucoseaminidase (NAG), were examined using conventional clearance techniques after three days of treatment with FK 506. RESULTS: FK 520 treatment induced a pronounced and dose-dependent decrease in PT Na+,K+-ATPase activity. This effect was completely reversed by the competitive FK 520 antagonist, L 685 818, indicating that the effect was dependent on inhibition of calcineurin. To test whether the FK 520-induced decrease in Na+, K+-ATPase activity was mediated by enhanced beta-adrenoceptor signaling, the FK 520 effect was examined in rats pretreated with a beta-adrenoceptor antagonist (propranolol) or rats subjected to renal denervation. Both of these procedures prevented the FK 520-induced decrease in Na+,K+-ATPase activity. Thus, during FK 520 treatment, renal sympathetic nerves mediate an inhibitory effect on PT Na+,K+-ATPase activity via beta adrenoceptors. Propranolol pretreatment also prevented FK 506-induced decreases in GFR and urinary excretion of NAG, an index of PT dysfunction. CONCLUSIONS: The results support the hypothesis that the net effect of the neurotransmitter NE on Na+,K+-ATPase activity is dependent on the balance between the alpha- and beta adrenergic signaling pathways and suggest that agents that interfere with these pathways may, by altering the activity of tubular Na+,K+-ATPase, also alter the function of the renal tubular epithelial cell. PMID- 10469370 TI - Presence of fructose transporter GLUT5 in the S3 proximal tubules in the rat kidney. AB - BACKGROUND: Fructose is a nutrient as well as a potent agent for the formation of advanced glycation end product in diabetes. GLUT5 is a facilitated-diffusion fructose transporter expressed in the small intestine and kidney. Previous reports on the localization of GLUT5 by in situ hybridization and immunohistochemistry were controversial. METHODS: The expression of GLUT5 was checked by reverse transcription-polymerase chain reaction and Southern blotting and immunoblotting analyses. Localization of GLUT5 was visualized by high resolution immunofluorescence and immunogold electron microscopy. RESULTS: We were able to confirm the expression of GLUT5 in the kidney. GLUT5 was predominantly present in the outer stripe of the outer medulla, where it was localized in the S3 proximal tubule cells. Double labeling with phalloidin showed that GLUT5 was localized in the brush border of the S3 proximal tubule cells. Ultrastructural examination revealed that GLUT5 was present along the plasma membrane of the apical microvilli. CONCLUSION: GLUT5 is present at the apical plasma membrane of S3 proximal tubule cells and may serve as the transporter of fructose. PMID- 10469371 TI - Intrarenal distribution of the colonic H,K-ATPase mRNA in rabbit. AB - BACKGROUND: Evidence suggests that the colonic H,K-ATPase isoform is expressed in the kidney and that a mRNA species highly homologous to the rat and guinea pig HKalpha2 is expressed in the cortical collecting duct (CCD) of the rabbit. The goals of this study were to determine if this mRNA is the rabbit homologue of HKalpha2 or a novel isoform and to determine intrarenal distribution of the HKalpha2 mRNA in rabbit. METHODS: 5'-RACE and Dye Deoxy Terminator chemistry were used to determine the full-length sequence of the rabbit HKalpha2 mRNA. The intrarenal distribution of HKalpha2 mRNA was determined in microdissected nephron segments, connecting tubule (CNT), and CCD cells isolated by immunodissection, as well as in the three cell types of the CCD. Principal cells and alpha- and beta intercalated cells were isolated by fluorescence-activated cell sorting. HKalpha2 mRNA levels were determined by quantitative reverse transcription-polymerase chain reaction (RT-PCR) or single-nephron RT-PCR (SN-RTPCR). RESULTS: The full length sequence of the rabbit kidney HKalpha2 mRNA was determined. This transcript is identical to the one expressed in rabbit distal colon. In microdissected nephron segments, strong HKalpha2 amplicons were present in the CNT, CCD, and outer medullary collecting duct (OMCD), whereas no signal was detected in the proximal tubule, distal convoluted tubule, think ascending limb, and inner medullary collecting duct. Roughly comparable levels of HKalpha2 mRNA were present in all three CCD cell types, and the highest levels were observed in a subpopulation most likely corresponding to CNT cells. CONCLUSIONS: These results suggest that the HKalpha2 mRNA is expressed in rabbit collecting duct is identical in size and sequence to the one expressed in rabbit distal colon. HKalpha2 mRNA in the rabbit kidney is selectively expressed in the CNT, CCD, and OMCD, and all three collecting duct subtypes express its mRNA. PMID- 10469373 TI - Lipoprotein(a) and apolipoprotein(a) isoforms and proteinuria in patients with moderate renal failure. AB - BACKGROUND: Atherosclerotic diseases are a major cause of death in patients with renal failure. Increased serum concentrations of lipoprotein(a) [Lp(a)] have been established as a genetically controlled risk factor for these diseases and have been demonstrated in patients with moderate renal failure, suggesting that this lipoprotein contributes to the increased cardiovascular risk seen in these patients. Variable alleles at the apolipoprotein(a) [apo(a)] gene locus are the main determinants of the serum Lp(a) level in the general population. The purpose of this study was to investigate apo(a) isoforms in patients with moderate renal failure and mild proteinuria (less than 1.0 g/day). METHODS: In 250 consecutive subjects recruited at a hypertension clinic, we assessed the renal function by 24 hour creatinine clearance, proteinuria, and microalbuminuria, as well as the prevalence of atherosclerotic disease, and we also measured apo(a) isoforms, serum albumin, and Lp(a) concentrations. RESULTS: Moderate impairment of renal function (creatinine clearance, 30 to 89 ml/min per 1.73 m2 of body surface area) was found in 97 patients. Lp(a) levels were significantly greater in patients with moderate renal failure (21.7+/-23.9 mg/dl) as compared with patients with normal renal function (15.6+/-16.4 mg/dl, P<0.001), and an inverse correlation was observed between log Lp(a) and creatinine clearance (r = -0.181, P <0.01). However, no difference was found in the frequency of low molecular weight apo(a) isoforms between patients with normal (25.5%) and impaired (27.8%) renal function. Only patients with the smallest size apo(a) isoforms exhibited significantly elevated levels of Lp(a), whereas the large-size isoforms had similar concentrations in patients with normal and impaired renal function. No significant relationship was found between serum Lp(a) and proteinuria. Clinical and laboratory evidence of one or more events attributed to atherosclerosis was found in 9.8% of patients with normal renal function and 25.8% of patients with moderate renal failure (P<0.001). CONCLUSIONS: These results indicate that renal failure per se or other genes beside the apo(a) gene locus are responsible for the elevation of serum Lp(a) levels in patients with moderate impairment of renal function. The elevation of Lp(a) levels occurs independently of the level of proteinuria and may contribute to the risk for atherosclerotic disease in these patients. PMID- 10469372 TI - Renin-angiotensin blockade lowers MCP-1 expression in diabetic rats. AB - BACKGROUND: Glomerular macrophage accumulation in diabetes implicates monocyte recruitment mechanisms in the pathogenesis of diabetic nephropathy. To test the hypothesis that overexpression of monocyte chemoattractant protein-1 (MCP-1), a monocyte chemoattractant, is attenuated by renin-angiotensin system (RAS) inhibition, we assessed expression of genes regulating monocyte transmigration in the glomeruli of diabetic rats. METHODS: Competitive reverse transcription polymerase chain reaction (RT-PCR) was used to semiquantitate mRNA expression in glomeruli harvested by sieving at serial intervals after the induction of diabetes by streptozotocin in Munich-Wistar rats. Although subject to limitations, competitive RT-PCR provides an objective measure suited to the minute quantities of RNA extractable from glomerular isolates. RESULTS: Time dependent elevation of MCP-1 expression was dramatically suppressed by treatment with the angiotensin-converting enzyme inhibitor enalapril or the AT1 receptor antagonist candesartan, and was closely associated with effects on proteinuria and glomerular macrophage number. By contrast, no sustained suppression of the cell adhesion molecules intercellular adhesion molecule-1 or vascular cell adhesion molecule-1 or the classic MCP-1 stimulators tumor necrosis factor-alpha or interleukin-1beta followed RAS inhibition, and suppression of transforming growth factor-beta1 expression was transient. CONCLUSION: These data suggest that glomerular macrophage recruitment in experimental diabetes is largely determined by angiotensin-stimulated MCP-1 expression. We conclude that the RAS is an important regulator of local MCP-1 expression, either directly or through glomerular hemodynamic effects, and that our data strongly implicate macrophage recruitment and activation in the pathogenesis of early diabetic glomerular injury. PMID- 10469374 TI - Role of hypoalbuminemia and hypocholesterolemia as copredictors of mortality in acute renal failure. AB - Role of hypoalbuminemia and hypocholesterolemia as co-predictors of mortality in acute renal failure. BACKGROUND: Hypoalbuminemia (LA) and hypocholesterolemia (LC) have been reported to portend high mortality in both older patients and in patients with end-stage renal disease. Even though low levels have been reported in critically ill patients, they have not been clearly defined as predictors of mortality in acute renal failure (ARF). The impact of LA and LC on mortality in ARF is evaluated in this study. METHODS: We conducted a computer-assisted three year retrospective review of all cases of de novo ARF seen at an inner city tertiary-care facility. One hundred cases met the criteria for inclusion in the study. We employed both univariate and multivariate logistic regression models to estimate the relative risks (RR) and 95% confidence intervals (CI) of mortality associated with several variables. RESULTS: Predictors associated with a high risk of death identified in this study include LC < or = 150 mg/dl (< or = 3.9 mmol/liter; RR, 7.4; CI, 2.7 to 20.3), LA < or =35 g/liter (RR, 5.0; CI, 1.9 to 13.2), sepsis (RR, 9.4; CI, 3.7 to 23.9), mechanical ventilation (RR, 10.8; CI, 2.8 to 41.0), oliguria (RR 17.0; CI, 6.2 to 46.6), and multisystem organ failure (RR 24.7; CI, 10.3 to 59.1). The overall gross mortality was 39%, but mortality among intensive care unit patients was 82%. Survival was 82% among patients with serum albumin >35 g/liter versus 48% among those with serum albumin < or =35 g/liter (chi2 = 11.9, P = 0.0006). Similarly, survival was higher among patients with cholesterol >150 mg/dl (>3.9 mmol/liter) than those whose levels were < or =150 mg/dl (< or =3.9 mmol/liter; 85 vs. 44%, ch 17.3, P<0.0001). Significant association between LA and LC was observed (R = 0.4, P<0.0001). Age, gender, level of plasma creatinine, and underlying chronic medical conditions were not predictive of mortality. CONCLUSION: Survival in ARF is significantly altered by the levels of albumin and cholesterol. Because both LC and LA can be cytokine mediated, their presence in ARF should be considered ominous. PMID- 10469375 TI - Homocysteine and methionine metabolism in ESRD: A stable isotope study. AB - BACKGROUND: Hyperhomocysteinemia has a high prevalence in the end-stage renal disease (ESRD) population, which may contribute to the high cardiovascular risk in these patients. The cause of hyperhomocysteinemia in renal failure is unknown, and therapies have not been able to normalize plasma homocysteine levels. Insight into methionine-homocysteine metabolism in ESRD is therefore necessary. METHODS: Using a primed, continuous infusion of [2H3-methyl-1-13C]methionine, we measured whole body rates of methionine and homocysteine metabolism in the fasting state in four hyperhomocysteinemic hemodialysis patients and six healthy control subjects. RESULTS: Remethylation of homocysteine was significantly decreased in the hemodialysis patients: 2.6+/-0.2 (SEM) vs. 3.8+/-0.3 micromol. kg(-1)x hr(-1) in the control subjects (P = 0.03), whereas transsulfuration was not 2.5+/-0.3 vs. 3.0+/-0.1 micromol. kg(-1) x hr(-1) (P = 0.11). The transmethylation rate was proportionally and significantly lower in the ESRD patients as compared with controls: 5.2+/-0.4 vs. 6.8+/-0.3 micromol. kg(-1) x hr(-1) (P = 0.02). Methionine fluxes to and from body protein were similar. CONCLUSIONS: The conversion of homocysteine to methionine is substantially (approximately 30%) decreased in hemodialysis patients, whereas transsulfuration is not. Decreased remethylation may explain hyperhomocysteinemia in ESRD. This stable isotope technique is applicable for developing new and effective homocysteine-lowering treatment regimens in ESRD based on pathophysiological mechanisms. PMID- 10469376 TI - A new dimension to the Barker hypothesis: low birthweight and susceptibility to renal disease. AB - BACKGROUND: There is an epidemic of renal failure among Aborigines in the Australia's Northern Territory. The incidence is more than 1000 per million, and is doubling every three to four years. We evaluated the relationship of birthweight to renal disease in adults in one high-risk community. METHODS: We screened more than 80% of people in the community for renal disease, using the urine albumin/creatinine ratio (ACR, g/mol) as the marker, and reviewed records for birthweights. RESULTS: Birthweights were available with increasing frequency for people born after 1956. In 317 adults aged 20 to 38 years at screening, the mean birthweight (SD) was 2.712+/-0.4 kg, and 35% had been low birthweight (LBW, less than 2.5 kg). Birthweight was positively correlated with body mass index (BMI), blood pressure, and diabetes rates, but was inversely correlated with ACR. The odds ratio for overt albuminuria in LBW persons compared with those of higher birthweights was 2.82 (CI, 1.26 to 6.31) after adjusting for other factors, and LBW contributed to an estimated 27% (CI, 3 to 45%) of the population-based prevalence of overt albuminuria. Multivariate models suggest that increasing BMI and blood pressure and decreasing birthweight act in concert to amplify the increases in ACR that accompany increasing age. CONCLUSIONS: LBW contributes to renal disease in this high-risk population. The association might be mediated through impaired nephrogenesis caused by intrauterine malnutrition. The renal disease epidemic in Aborigines may partly be the legacy of greatly improved survival of LBW babies over the last four decades. Disease rates should eventually plateau as birthweights continue to improve, if postnatal risk factors can also be contained. PMID- 10469377 TI - Serum malondialdehyde and prevalent cardiovascular disease in hemodialysis. AB - BACKGROUND: Oxidative stress has been proposed as a mechanism by which the accelerated rate of cardiovascular disease (CVD) observed in maintenance hemodialysis (HD) patients may be explained. This study examined the effects of HD and CVD on serum malondialdehyde (MDA) levels as a marker of oxidative stress in HD patients with and without prevalent CVD. Serum MDA levels and CVD prevalence in HD were modeled. METHODS: Serum MDA was determined using spectrophotometry in HD patients (N = 76, 53 men and 23 women, mean age 63.8 years) immediately prior to and at the conclusion of one midweek HD treatment. Traditional CVD risk factors, including serum lipids, lipoproteins, apolipoproteins, and fibrinogen, were also measured, as were serum chemistry and dialysis adequacy. RESULTS: Mean serum MDA levels were significantly elevated in HD patients with prevalent CVD compared with those without, whereas serum lipoprotein and plasma fibrinogen levels did not differ between the two groups. Patients in the highest compared with the lowest tertile of postdialysis MDA were nearly four times as likely to have prevalent CVD, and serum MDA was the single strongest predictor of prevalent CVD in this patient population. CONCLUSIONS: These findings indicate the presence of oxidative stress in HD patients, and are consistent with the theory of oxidative stress as a factor in accelerated CVD in this population. PMID- 10469378 TI - Bone mineral density and biochemical markers of bone turnover in patients with predialysis chronic renal failure. AB - BACKGROUND: Metabolic bone disease might commence early in the course of renal failure. This study therefore examined the frequency and severity of the skeletal changes in predialysis chronic renal failure by measurements of bone mineral density (BMD), biochemical markers of bone turnover (osteocalcin, bone-specific alkaline phosphatase, carboxy terminal propeptide of type I collagen, and carboxy terminal telopeptide of type I collagen), parathyroid hormone (PTH), ionized calcium (Ca++), phosphate (P), and vitamin D metabolites. METHODS: The study was performed in 113 patients (male/female: 82/31) with chronic renal diseases [mean glomerular filtration rate (GFR) of 37 ml/min] and in 89 matched, normal control subjects. RESULTS: The patients had significantly (P<0.05) reduced BMD in the spine (-6.3%), the femur (-12.1%), the forearm (-5.7%), and the total body ( 4.2%) as compared with the control subjects. Dividing the patients into quartiles according to GFR revealed that BMD decreased with the gradual decline in renal function at all the measured skeletal sites, but was most pronounced in the femur: 0.63+/-0.03, 0.74+/-0.02, 0.77+/-0.02, and 0.82+/-0.03 g/cm2 in each quartile from lowest to highest GFR compared with 0.82+/-0.02 g/cm2 in the control group (P<0.0001). All of the measured bone markers showed increasing plasma levels with the more advanced stages of renal failure. Serum PTH and serum P levels increased, whereas serum Ca++ and 1,25-dihydroxyvitamin D decreased. BMD Z-scores of the femur and of the forearm correlated to the biochemical markers and to PTH (P<0.05 to P<0.0001). The biochemical markers all showed strong correlations to PTH, also when corrected for the effect of the decline in GFR (r = 0.40 to 0.92, P<0.01 to P< 0.0001). CONCLUSION: Skeletal changes are initiated at an early stage of chronic renal failure, as estimated from reduced BMD and elevated levels of PTH and from the biochemical markers of both bone formation and bone resorption. PMID- 10469379 TI - Transplantation for primary hyperoxaluria in the United States. AB - BACKGROUND: Transplantation (TX) has become an acceptable treatment for renal failure in primary hyperoxaluria (PH). We have analyzed data from three U.S. sources to estimate the success or failure of different modes of management in PH patients. METHODS: The United States Renal Data System (USRDS) tapes provided coded medical record data, with PH assigned to 235 patients from 1974 to 1996. Another 45 patients were found from USRDS hospitalization records. We limited patients to those developing end-stage renal disease at <55 years of age after 1984 (95 PH patients). The North American Pediatric Renal Transplantation Cooperative Study (NAPRTCS) identified 34 (11 new) PH patients, and the United Network for Organ Sharing (UNOS) database identified PH in 34 (16 new, 5 more in both UNOS and NAPRTCS) patients. These secondary sources were used to correct some data from the USRDS and to add 32 more patients, with a total of 128 PH patients. Considering kidney TX (KTX) prior to combined kidney/liver TX (K/LTX) as a separate record for some calculations, the total "cases" were 138. RESULTS: By life table analysis, the 94 total TX patient survival was better than for the 34 NoTX patients (P<0.001). The 52 KTX patients' survival was better than either the 32 primary K/LTX (P<0.001) or the 10 K/LTX that following KTX (P<0.001). The 62 KTX cases' survival was better than the 42 K/LTX cases (P<0.005), which did not differ from the 34 NoTX (P<0.67). The overall survival of these 62 KTX patients was 76%. The survival of 42 K/LTX was 69%, and the survival of 34 NoTX patients was 44%. Kidney graft life table projected survival curves for TX patients did not differ between K/LTX (56% at 6 years) and isolated KTX (51% at 6 years, 35% at 10 years, P<0.91). CONCLUSION: KTX offers better patient survival in the United States then either K/LTX or NoTX. Graft survival does not differ between KTX and K/LTX. Because K/LTX can still follow a failed KTX, isolated living related donor KTX is still a reasonable first option for PH type 1 if a strictly managed protocol is followed. PMID- 10469380 TI - Peripheral donor leukocytes prolong survival of rat renal allografts. AB - BACKGROUND: The development of strategies to enhance the survival of transplanted organs and to potentially lower or even discontinue immunosuppressive therapy would represent a significant advancement in post-transplant patient care. METHODS: We studied the effect of pretransplant infusion of donor leukocytes alone or in combination with a short course of cyclosporine on the long-term outcome of a rat model of kidney allograft. RESULTS: A single intravenous infusion of donor peripheral blood leukocytes (100x10(6) cells) from Brown-Norway (BN) rats into major histocompatibility complex (MHC) incompatible Lewis recipients largely failed to prolong kidney allograft viability from the same donor transplanted 60, 40, or 30 days after cell infusion. A short course of cyclosporine (per se, unable to prolong graft survival) was started at the same day of donor leukocyte infusion, but instead was able to prolong the survival of the BN kidney transplant-performed 40 days later-but not of a Wistar Furth (WF) third party, with some animals even developing tolerance. A mixed lymphocyte reaction of host cells from long-term surviving rats to BN stimulator cells was significantly reduced as compared with controls. Donor BN DNA was detected in the peripheral blood of Lewis rats until day 40 after BN leukocyte infusion. Microchimerism persisted (60 to 70 days post-transplant) in most long-term graft recipients. Reducing the time interval between donor leukocyte infusion and subsequent kidney transplant to 10 days still prolonged graft survival. Donor peripheral blood mononuclear cells, but not polymorphonuclear cells, in the leukocyte preparation contributed to prolong kidney allograft survival. CONCLUSIONS: Pretransplant donor leukocyte infusion under the appropriate conditions can tip the immune balance toward improved graft acceptance. This result could be relevant to the achievement of donor-specific tolerance of the graft with the maintenance of an intact response to third-party antigens. PMID- 10469381 TI - Factors determining the percentage of hypochromic red blood cells in hemodialysis patients. AB - Factors determining the percentage of hypochromic red blood cells determines iron status in hemodialysis patients. BACKGROUND: Determination of the percentage of hypochromic red blood cells (RBC; %HYPO) has been advocated as a sensitive index of functional iron deficiency during erythropoietin (EPO) therapy in hemodialyzed patients. METHODS: The significance of %HYPO in chronic renal failure was evaluated in 64 chronically hemodialyzed patients. The linear correlation was determined between %HYPO and 13 variables, including age, sex, weight, C-reactive protein (CRP), ferritin, transferrin (Tf), Tf saturation, soluble Tf receptor (sTfR), serum iron (SI), urea, parathormone, dialysis dose (Kt/V), dose of EPO administered (EPO), and absolute reticulocyte count. Multiple regression analyses were then performed to select the parameters that jointly provide the best prediction of %HYPO. RESULTS: Univariate analysis showed significant correlations between %HYPO and iron parameters (sTfR, Tf saturation, SI, and ferritin, in decreasing order), EPO, reticulocyte count, and CRP. Multivariate analysis yielded an equation showing that the variation of %HYPO is essentially associated with the combined changes in sTfR, CRP, and EPO dosage. CONCLUSIONS: %HYPO is a meaningful and inexpensive parameter that reflects the integrated effects of iron stores, inflammation, and erythropoietic stimulation on iron availability in hemodialyzed patients. Among iron exchange parameters, sTfR is the best predictor of %HYPO, followed by Tf saturation, SI, and ferritin. PMID- 10469382 TI - GBV-C/HGV-RNA in serum and peripheral blood mononuclear cells in hemodialysis patients. AB - BACKGROUND: Hemodialysis patients are at high risk of hepatitis B, C, and G virus infection. The prevalence of GBV-C/HGV-RNA was analyzed in serum and peripheral blood mononuclear cells (PBMCs) from 52 hemodialysis patients. METHODS: GBV-C/HGV RNA detection was performed by reverse transcription-polymerase chain reaction (RT-PCR) with primers of 5'-noncoding (5'-NC) and NS3 regions of the GBV-C/HGV genome. To increase sensitivity, serum samples were ultracentrifuged prior to the RT-PCR to concentrate the viral particles. The amplified products from 20 serum and 5 peripheral blood mononuclear cells (PBMC) samples were sequenced. RESULTS: GBV-C/HGV-RNA was detected in sera of 9 (17%) and in PBMCs of 30 (58%) patients. After serum ultracentrifugation, GBV-C/HGV-RNA was positive in 20 (95%) of the patients, with GBV-C/HGV-RNA only in PBMCs. Thus, GBV-C/HGV-RNA was detected in serum and PBMCs from 29 (56%) patients, four of whom had antibodies against GBV C/HGV E2 protein (anti-HGE2); one patient (2%) had GBV-C/HGV-RNA only in PBMCs, but was anti-HGE2 positive. Seven (32%) patients who did not have GBV-C/HGV-RNA were anti-HGE2 positive. The nucleotide sequence homology between serum samples from the patients who were GBV-C/HGV-RNA positive after ultracentrifugation, and paired serum and PBMCs from five of them, ranged from 90 to 96% and from 92 to 98%, respectively. CONCLUSIONS: We found a high prevalence of GBV-C/HGV-RNA in serum and PBMC samples from hemodialysis patients. Whether or not this finding can be extended to other populations requires further study. PMID- 10469383 TI - Ultrafiltration method for measuring vascular access flow rates during hemodialysis. AB - BACKGROUND: The vascular access blood flow rate (QA) has been shown to be an important predictor of vascular access failure; therefore, the routine measurement of QA may prove to be a useful clinical method of vascular access assessment. METHODS: We have developed a new ultrafiltration (UF) method for determining QA during HD from changes in arterial hematocrit (H) after abrupt changes in the UF rate with the dialysis blood lines in the normal (DeltaHn) and reverse (DeltaHr) configurations. This method accounts for cardiopulmonary recirculation and requires neither intravenous saline injections nor accurate knowledge of the dialyzer blood flow rate. Clinical studies were conducted in 65 chronic HD patients from three different dialysis programs to compare QA determined by the UF method with that determined by saline dilution using an ultrasound flow sensor. RESULTS: Arterial H increased (P<0.0001) after abrupt increases in the UF rate when the lines were in the normal and reverse configurations. An increase in the UF rate from the minimum setting to 1.8 liter/hr resulted in a DeltaHn of 0.3+/-0.2 (mean +/- SD) H units and a DeltaHr of 1.6+/-1.0 H units. Q(A) values determined by the UF method (1050+/-460 ml/min) were 16+/-25% higher (P<0.001) than those determined by saline dilution (950+/ 440 ml/min); the calculated QA values by the UF and saline dilution methods correlated highly with each other (R = 0.92, P<0.0001). The average coefficient of variation for duplicate measurements of QA determined by the UF method in a subset of these patients (N = 21) was approximately 10% when assessed in either the same dialysis session or consecutive sessions. CONCLUSIONS: The results from this study show that changes in arterial H after abrupt changes in the UF rate can be used to assess Q(A). PMID- 10469384 TI - Body weight-for-height relationships predict mortality in maintenance hemodialysis patients. AB - BACKGROUND: Protein-energy malnutrition is a strong predictor of mortality in maintenance hemodialysis (MHD) patients. This association has generally been described for serum chemistry measures of protein-energy malnutrition. We hypothesized that body weight-for-height relationships also predict survival in MHD patients. METHODS: During the last three months of 1993, data were obtained on 12,965 men and women concerning clinical characteristics (height, postdialysis weight, age, gender, race, and presence or absence of diabetes mellitus) and laboratory measurements (predialysis serum albumin, creatinine and cholesterol, and the urea reduction ratio). Patient survival during the next 12 months was evaluated retrospectively. RESULTS: In comparison to values for normal Americans determined from the National Health and Nutrition Evaluation Survey II data, weight-for-height relationships tended to be slightly lower than normal in African American men and women and Caucasian men undergoing MHD and were normal or slightly greater in the taller Caucasian women. In both men and women, the mortality rate decreased progressively as the patients' weight-for-height increased. MHD patients who weighed more than normal had the lowest mortality rates. After adjustment for clinical characteristics and laboratory measurements, the inverse relationship between mortality rates and weight-for-height percentiles was still highly significant for patients within the lower 50th percentile of body weight-for-height. Serum albumin correlated directly with weight-for-height in patients in the lower 50th percentile of weight-for-height. Serum creatinine and cholesterol correlated directly with weight-for-height in the entire population of men and women. In contrast, the urea reduction ratio was inversely correlated with weight-for-height. CONCLUSIONS: These data indicate that weight-for-height is a strong predictor of 12-month mortality in male and female MHD patients. Multivariate analyses indicate that body weight-for-height is an independent predictor of higher mortality in those patients who are in the lower 50th percentile for this measurement. PMID- 10469385 TI - Quest for postdialysis urea rebound-equilibrated Kt/V with only intradialytic urea samples. AB - BACKGROUND: Postdialysis urea rebound (PDUR) is a cause of Kt/V overestimation when it is calculated from predialysis and the immediate postdialysis blood urea collections. Measuring PDUR requires a 30- or 60-minute postdialysis sampling, which is inconvenient. Several methods had been devised for a reasonable approach to determine PDUR-equilibrated Kt/V in short dialysis without the need for a delayed sample. The aim of our study was to compare these different Kt/V methods during the longer eight-hour hemodialysis sessions, and to determine the optimum intradialytic urea sample time that fits best with PDUR. METHODS: The study included 21 patients (mean age 71.9 years) who were hemodialyzed for 60+/-60 months at three times eight hours weekly, using bicarbonate dialysate and cellulosic membranes. Blood urea samples were obtained at onset, and then at 17, 33, 50, 66, 75, 80, 85, and 100% of the dialysis session times, after 30 seconds of low flow, and then at 60-minutes postdialysis. All patients had a meal during dialysis. We compared four different formulas of Kt/V [(a) Kt/V-Smye with a 33% dialysis time urea sample, (b) two-pool equilibrated eKt/V, (c) Kt/V-std (Daugirdas-2) obtained with an immediate postdialytic sample, and (d) the different intradialytic urea samples for Kt/V (50, 66, 75, 80, and 85% of dialysis time)] with the equilibrated 60-minute PDUR Kt/V (Kt/V-r-60) formula as the reference method. RESULTS: The mean PDUR was 17.2+/-9%, leading to an overestimation of Kt/V-std by 12.2%. Kt/V-r-60 was 1.68+/-0.34. Kt/V-std was 1.88+/-0.36 (Delta = 12.2+/-4.8%, r = 0.8). eKt/V was 1.77+/-0.3 (Delta = 5+/-5%, r = 0.96), and Kt/V-Smye was 1.79+/-0.47 (Delta = 5.2+/-14%, r = 0.9). The best time for the intradialytic sampling was 80% (that is, at 6 hr and 24 min). The Kt/V-80 was 1.64+/-0.3 and was best fitted with Kt/V-r-60 (Delta = -1.8+/-8%, r = 0.91). The mean intradialytic urea evolution showed a three-exponential rate, in discrepancy with the two-exponential rate theoretical model. CONCLUSIONS: These results confirm that a significant postdialysis rebound exists in an eight-hour dialysis. An intradialytic urea sample taken at 80% of the total session time permits an estimation of the 60-minute Kt/V-rebound without the necessity of taking a delayed sample, with better accuracy than eKt/V or especially Kt/V-Smye. This may be related to a particular urea kinetics curve on the longer dialysis duration, which needs to be studied further. PMID- 10469386 TI - A role for leptin in glomerulosclerosis? PMID- 10469387 TI - FGF-2: specific activity in kidney? PMID- 10469388 TI - Evidence of lectin complement pathway activation in poststreptococcal glomerulonephritis. PMID- 10469389 TI - HIV-associated nephropathy. PMID- 10469390 TI - Lobular involvement: prognostic indicator for invasive ductal carcinoma of the breast. AB - One of the striking microscopic appearances of breast carcinoma is malignant transformation of the epithelium of an entire lobule, although the lobular architecture is still preserved. We termed this pathological feature 'lobular involvement'. An investigation was carried out to determine whether lobular involvement would be a useful prognostic indicator for invasive ductal carcinoma. One hundred and fifty cases of invasive ductal carcinoma were included in this study. The lobular involvement, as well as the conventional prognostic factors, was evaluated to find out the correlation with the overall and relapse-free survivals by univariate and multivariate analyses. The lobular involvement was identified in 31 of 150 patients (20.6%). Of the patients with lobular involvement, two had experienced recurrence (6.5%) and one had died (3.2%). The lobular involvement and lymph node metastases were significantly and independently correlated with the overall and relapse-free survivals. The presence of lobular involvement was statistically correlated with the favorable outcome. Lobular involvement can be assumed as a new prognostic indicator for invasive ductal carcinoma. PMID- 10469391 TI - Thymidine phosphorylase expression in human colorectal mucosa, adenoma and carcinoma: role of p53 expression. AB - The enzyme, thymidine phosphorylase (dThdPase) is identical to platelet-derived endothelial cell growth factor (PD-ECGF), which acts as a potent angiogenic factor. The present study immunohistochemically examined the expression of dThdPase in human colorectal mucosa, adenomas and carcinomas, as well as six cultured colorectal carcinoma cell lines, in terms of intratumoral microvessel density (IMVD) and P53 expression. Thymidine phosphorylase was observed in lymphocytes, fibroblasts and macrophages, as well as smooth muscle cells and Schwann cells in the peripheral nerve fibers. The dThdPase-positive stromal cells apparently outnumbered the normal epithelial cells, adenoma and carcinoma cells with dThdPase. Weak but obvious cytoplasmic immunoreactivity was noted in a few normal colonic epithelia, predominantly the upper surface area, while a few adenoma cells showed weak nuclear immunostaining for dThdPase in six (24%) of the 25 colonic adenomas. Expression of dThdPase was noted in 33 (73.3%) of the 45 Dukes A and B, 14 (51.9%) of the 27 Dukes C and 14 (56.0%) of the 25 Dukes D carcinomas. The mean IMVD was 84.0 +/- 26.2 in the 36 dThdPase-negative carcinomas and 97.9 +/- 31.6 in the 61 dThdPase-positive carcinomas, the value being significantly higher in the latter group (P < 0.05). The frequency of dThdPase expression was significantly lower in the P53-negative carcinomas than in the positive carcinomas (P < 0.05). Western blot analysis showed the highest expression of dThdPase in LoVo carrying the wild-type p53 gene, followed by Colo201, Colo320, DLD-11 and WiDr carrying the mutated gene. These results indicate that: (i) the main source of dThdPase is stromal cells, including lymphocytes and macrophages in both colorectal normal and carcinoma tissues; (ii) dThdPase may take part in the induction of intratumoral microvessels, regardless of tumor stage; and (iii) expression might be modulated by not only P53 but also other molecules. PMID- 10469392 TI - Localization of prostate-specific antigen-like immunoreactivity in human salivary gland and salivary gland tumors. AB - Immunoreactivity of prostate-specific antigen (PSA), a kallikrein-like enzyme present in the seminal plasma, was demonstrated by indirect immunoperoxidase staining using a PSA antiserum in the apical cytoplasm along the luminal border of small-sized duct epithelial cells of the major salivary (parotid and submandibular) gland of both sexes (56/56, 100%). No PSA-like immunoreactivity was seen in large-sized duct epithelial cells and acinar cells. Minor salivary gland ducts were negative. When inflammatory and atrophic changes were observed, ductal expression of PSA-like immunoreactivity was decreased (12/37, 32%) and the site of intracellular localization often became diffusely cytoplasmic. The immunoreactivity was absorbed by human seminal plasma. Immunoreactivities of prostatic acid phosphatase and sex hormone receptors were undetectable in the salivary gland. Twenty-nine (34%) of 86 salivary gland tumors with ductal differentiation were immunoreactive for PSA mainly in the cytoplasm. A PSA monoclonal antibody ER-PR8 detected immunoreactivity in the prostate but not in the salivary glands or their tumors. Prostate-specific antigen-like immunoreactivity in small-sized (intercalated) duct epithelial cells of the major salivary gland and their tumors may be due to cross-reactivity of the antiserum with kallikrein-like substances. PMID- 10469393 TI - Expressions of cyclin E, A, and B1 in Hodgkin and Reed-Sternberg cells: not suppressed by cyclin-dependent kinase inhibitor p21 expression. AB - p21 Is involved in the control of the mammalian cell cycle through the binding and inhibition of cyclin-dependent kinases. The cyclins are dependent on the phases of the cell cycle, and divided into two classes: mitotic cyclins (A, B1, B2) and G1 cyclins (C, D1, D2, D3, E). The product of the p21 gene is a potent downstream effector of the p53 tumor-suppressor gene function. The Hodgkin and Reed- Sternberg (H & RS) cells in Hodgkin's disease are reported to frequently express p53, p21, and nuclear proliferative activity (Ki-67). To clarify the relationship of p21, p53 and cyclins, we performed the immunohistochemistry of p53, p21, Ki-67, cyclin D1, cyclin E, cyclin A and cyclin B1, using 11 cases with Hodgkin's disease. In addition, we performed p53 gene sequencing of exon 5-8, and in situ hybridization of Epstein-Barr virus (EBV) EBER-1 region, whose products have reported to induce the expression of cyclin D. In this study, in all cases, Ki-67 was expressed in almost all H & RS cells, and p53 and p21 were expressed in H & RS cells. No p53 gene mutations were detected in any case, and p53 protein overexpression did not correlate with p53 gene mutations. The number of p21 positive H & RS cells was significantly related with that of the p53-positive cells. The cyclins E, A, B1 and D1 were also expressed in H & RS cells. Unexpectedly, the expression of the cyclins was not suppressed by p21 and p53 expression. In addition, the existence of EBV was not related to the expression of cyclins. It is considered that H & RS cells are, indeed, in cell cycle and commonly express the cell cyclins, and that the cell cycle of H & RS cells may not be specifically fixed in the G1, S, G2 or M phases. PMID- 10469394 TI - Intracellular localization of HSP73 and HSP90 in rat kidneys with acute lysosomal thesaurismosis. AB - We previously reported that HSP73 and HSP90, major chaperone proteins, accumulated within lysosomes of proximal tubular epithelial cells in rat kidneys with acute gentamicin nephropathy. In this study, we observed serial localization of HSP73 and HSP90 in rat kidneys with acute lysosomal thesaurismosis. Sprague Dawley rats received poly-D-glutamic acid (PDGA) (250 mg/kg per day) for 3 days, and developed acute lysosomal thesaurismosis of proximal tubular epithelial cells. The intracellular localization of HSP73 and HSP90 was examined by electron microscopy. We also compared the results with those of a non-chaperone protein, a renal isoform of argininosuccinate synthetase, which is an abundant enzyme in proximal tubular epithelial cells. After the PDGA exposure, HSP73 and HSP90 accumulated within enlarged lysosomes of proximal tubular epithelial cells. These accumulations started to appear from day 4 after the first PDGA administration, enlarged in size until day 14, and continued until day 19. Argininosuccinate synthetase also accumulated within the lysosomes, but the magnitude of this lysosomal accumulation was less than those of HSP73 and HSP90. Our findings demonstrated that HSP73 and HSP90 chaperone proteins specifically accumulated within lysosomes of proximal tubular epithelial cells during the course of PDGA induced acute lysosomal thesaurismosis. PMID- 10469395 TI - The role of Kupffer cells and regulation of neutrophil migration into the liver by macrophage inflammatory protein-2 in primary listeriosis in mice. AB - Depletion of mouse Kupffer cells and splenic macrophages following intravenous administration of liposome-entrapped clodronate severely reduced host resistance to primary infection with Listeria monocytogenes. Infection of clodronate-treated mice with a sublethal dose of L. monocytogenes resulted in death of the mice within 3 days. The macrophage depletion resulted in marked increases in bacterial growth in the liver and spleen, but not in other tissues. The proliferation of L. monocytogenes was observed in a large number of hepatocytes that underwent apoptosis. Infiltration of neutrophils in the liver and rapid formation of microabscesses were observed in the control mice after L. monocytogenes infection. However, there was less accumulation of neutrophils in the liver of Kupffer cell-depleted mice than in the control mice. Expression of macrophage inflammatory protein-2 (MIP-2) was enhanced in the livers of both the control and Kupffer cell-depleted mice after L. monocytogenes infection. MIP-2 was also induced in a murine hepatocyte cell line following L. monocytogenes infection. The administration of neutralizing anti-interleukin-8 receptor homolog antibody severely abrogated neutrophil infiltration into the Listeria-infected mouse liver. Anti-MIP-2 antibody moderately reduced neutrophil infiltration and microabscess formation in the liver. These findings indicate that Kupffer cells protect hepatocytes from L. monocytogenes infection and the resultant apoptosis. Moreover, MIP-2 and its related molecules produced by the infected hepatocytes regulate neutrophil infiltration and microabscess formation in primary listeriosis. PMID- 10469396 TI - Clinicopathological findings of virus-associated hemophagocytic syndrome in bone marrow: association with Epstein-Barr virus and apoptosis. AB - Non-neoplastic hemophagocytic syndrome (HPS), also called virus-associated hemophagocytic syndrome (VAHS), has been thought to be a distinct clinical entity. A spontaneous recovery is common, but the prognosis of Epstein-Barr virus (EBV)-associated VAHS is poor. However, the role of EBV has yet to be clearly elucidated. A retrospective study of the bone marrow of 30 cases, in which the diagnosis of non-neoplastic VAHS was clinicopathologically confirmed, was performed. We were unable to histologically confirm the presence of neoplastic lesions, especially lymphoma cell infiltration. Ten of the patients were children (aged less than 15 years) and young adults (aged under 20 years; median age, 10 years). Twenty patients were adults (aged over 21 years; median age, 48 years). Twelve of these patients died, while 18 showed a spontaneous recovery. We performed immunological staining and in situ hybridization (ISH) for EBV. To clarify the presence of apoptosis, an in situ apoptosis detection (tunnel) method was used. In situ hydridization showed an EBV-presence in 16 of the 30 patients. In addition, the EBV-presence was confined in the lymphocytes, especially T lymphocytes in double stainings. The number of EBV-infected cells varied; however, the EBV presence was associated with ages. Nine of the 10 children and young adults showed an EBV-presence, while EBV was detected in seven of the 20 adults. Especially in 10 patients aged over 49 years, no EBV was detected. According to the in situ apoptosis detection, apoptotic cells were increased in number and considered to be lymphoid cells, but not myeloid or histiocytic cells. Some apoptotic cells were phagocyted with histiocytes. Histologically, apoptosis may be one of the factors that induced phagocytosis. PMID- 10469397 TI - Aeromonas sobria infection with severe soft tissue damage and segmental necrotizing gastroenteritis in a patient with alcoholic liver cirrhosis. AB - A 49-year-old man, who had a 3-year history of liver dysfunction but had not been treated, was admitted to the hospital with a sudden onset of fever and generalized muscle pain. He subsequently developed generalized purpura with scattered hemorrhagic bullae of the skin and massive bloody stools. Aeromonas sobria was proven by culture of both blood and bullous fluid. In spite of the extensive treatment with antibiotics and other medications in the intensive care unit (ICU), the patient went into septic shock and died 2 days after admission. Pathological examination on autopsy revealed segmental necrotizing gastroenteritis with bacterial colonies and alcoholic liver cirrhosis, in addition to extensive severe soft tissue damage involving cellulitis and rhabdomyolysis and epidermolysis. Although the prognosis for Vibrio vulnificus infection with severe soft tissue damage in patients with liver cirrhosis, malignancy, diabetes mellitus or other pre-existing diseases is poor, the unfavorable progression of Aeromonas species, especially A. sobria infection is rare. This is thought to be the first report of an autopsied case. PMID- 10469398 TI - Epithelioid hemangioendothelioma with marked liver deformity and secondary Budd Chiari syndrome: pathological and radiological correlation. AB - A case of malignant epithelioid hemangioendothelioma of the liver in a 48-year old woman with severe portal hypertension and marked deformity of the liver is presented. This woman had a history of mild liver dysfunction since the age of 30 years, and abdominal distention, esophageal varices, splenomegaly and ascites since October 1996. Imaging examinations revealed liver deformity with severe atrophy of the left lobe and the anterior segment of the right lobe. Celiac arteriography showed narrowing and upward deviation of the proper hepatic artery, and occlusion of the left and right anterior hepatic arteries. Since March 1997, hepatic venography showed stenosis in the right hepatic vein truncus. Budd-Chiari syndrome was clinically diagnosed. She died in June 1997. The autopsy disclosed massive tumor embolism in the left and right anterior portal branches, few in the hepatic artery, and occlusion of the left and right anterior hepatic arteries. The extensive tumor embolism resulted in portal hypertension, and atrophy of the left lobe. The anterior segment of the right lobe was probably caused by the occlusion of both the hepatic arteries and the portal veins. The posterior segment of the right lobe, without massive tumor embolism in its portal branch, appeared hypertrophic. PMID- 10469399 TI - Langerhans cell histiocytosis of the stomach with atypical morphological features. AB - We report the unique case of a 52-year-old patient with atypical morphological features of gastric Langerhans cell histiocytosis. The man was admitted because of increasing upper abdominal pain and weight loss. The upper gastrointestinal endoscopy showed a submucous, hemispherical tumor of the stomach wall along the lesser curvature. The tumor was completely removed and the patient was discharged 11 days later. Two months later, he died at home. The cause of death is unknown, because the autopsy was denied by the relatives. Macroscopically, the stomach showed a 4.5 x 2.5 cm large, spherical tumor of the lesser curvature of the corpus, which infiltrated the perigastric omenta and lymph nodes, the distal pancreas and the glissonian liver capsule. The vast majority of neoplastic cells were intensely positive for S-100 and CD1a. Typical Birbeck granules could be identified in almost all cells. Cell nuclei analyzed by flow cytometry showed an aneuploid peak, a feature typically associated with malignant disease. PMID- 10469401 TI - Rectal adenocarcinoid with lymph node metastasis. AB - We describe a case of a rare variant of a rectal carcinoid tumor that showed mucous gland differentiation accompanied by a lymph node metastasis with a histological appearance similar to that of the primary site. The tumor consisted of a typical argyrophilic carcinoid component and of goblet cell glands. The carcinoid component was positive for neuron-specific enolase, chromogranin A and synaptophysin. The goblet cells stained positively with periodic acid-Schiff (PAS) and alcian blue, and expressed carcinoembryonic antigen, but were negative for neuroendocrine markers. This case suggests that carcinoid tumor can differentiate towards mucus glands, which can also be found in the metastatic site. PMID- 10469400 TI - Yolk sac tumor of the stomach with an adenocarcinomatous component: a case report with immunohistochemical analysis. AB - A 56-year-old male treated for a gastric yolk sac tumor with an adenocarcinomatous component is described. A mixed area of reticular and glandular neoplastic components was morphologically identified in this tumor. Immunohistochemically, the yolk sac tumor expressed alpha-fetoprotein (AFP), placental alkaline phosphatase (PLAP), and cytokeratin, but was negative for carcinoembryonic antigen (CEA). The adenocarcinoma was positive for CEA and cytokeratin, partially positive for PLAP, and negative for AFP. In the mixed area, the tumor cells were positive for cytokeratin, weakly expressed AFP and PLAP, and sporadically stained for CEA both in the reticular and glandular components. This area was identified as a transitional area of the yolk sac tumor and adenocarcinoma. These findings demonstrate that the yolk sac and adenocarcinomatous components are closely related. It also suggests that the tumor arose from multipotential neoplastic mucosal epithelial cells with both yolk sac and gastric mucosal phenotypes. PMID- 10469402 TI - A supernumerary ovary of the omentum with cystic change: report of two cases and review of the literature. AB - A supernumerary ovary is a rare gynecological anomaly. Particularly rare is the presence of cystic changes within the supernumerary ovary. We report two cases of neonates found to have a supernumerary ovary resembling an omental cyst. To the best of our knowledge, this report describes the first antenatal diagnosis of an omental cyst with a supernumerary ovary. To explain this unusual occurrence, it is suggested that an omental cyst becomes detached from the ovarian tissue and implants itself in the greater omentum, and that these supernumerary ovaries are of true embryologic origin, and not due to post-surgical or post-inflammatory implantation. PMID- 10469403 TI - Myoepithelioma of soft tissue. AB - A myoepithelioma occurred in the subcutaneous tissue of the right shoulder of a 28-year-old man. The well-demarcated nodular tumor (3. 0 x 2.8 cm) was located in the subcutaneous tissue with no adhesion to the deltoid muscle. The tumor was composed of a fascicular proliferation of spindle cells with variable amounts of stroma and showed areas of sheets of epithelioid cells. In most areas, the tumor cells had uniform nuclei, but pleomorphic epithelioid cells were focally present. Mitotic activity was three per 10 high-power fields. No ductular structure was found throughout the tumor. Immunohistochemical and ultrastructural studies confirmed the myoepithelial origin of the tumor cells. The occurrence of myoepithelioma in the subcutaneous tissue has been rarely reported. Even though the tumor showed no aggressive behavior on the 2-year follow-up, it is still too early to comment definitely on the behav- ior of myoepithelioma of the subcutaneous tissue. This case provides further information about soft tissue myoepithelioma. PMID- 10469404 TI - An autopsy case of primary nodal plasmacytoma associated with Sjogren's syndrome. AB - An 81-year-old man with a 1 year history of Sjogren's syndrome and hypergammaglobulinemic purpura was admitted because of high fever and lymphadenopathy. Primary nodal plasmacytoma was suggested from the microscopic and immunohistochemical findings of an inguinal lymph node biopsy specimen. Although chemotherapy achieved a moderate response, the patient died 2 months later from respiratory and cardiac failure. We herein, is described a rare autopsy case of primary nodal plasmacytoma associated with Sjogren's syndrome, in which the clinical course was progressive and Epstein-Barr virus-encoded RNA 1 was positive in a small number of neoplastic plasma cells, showing some difference from previously reported cases. PMID- 10469405 TI - Aberrant pancreatic tissue accompanied by heterotopic gastric mucosa in the gall bladder. AB - A 5 mm pancreatic heterotopia was incidentally identified in the gall-bladder with small cholesterol polyps following cholecystectomy in an asymptomatic 49 year-old female. Microscopically, inflammatory changes in the wall were minimal. The aberrant tissue consisted of acini and ducts but did not have islet formation. Heterotopic gastric mucosal tissue with a mixture of foveolar cells and pyloric glands was seen at the orifice of the pancreatic heterotopia. Islet peptide-immunoreactive endocrine cells were scattered among the pancreatic parenchyma, and the gastric-type gland contained cells positive for gastrin, somatostatin and glucagon. A noteworthy finding was the immunohistochemical detection of a single insulin-containing cell in the pyloric gland. PMID- 10469406 TI - Tinea pedis in children presenting as unilateral inflammatory lesions of the sole. AB - Tinea pedis is uncommon in prepubescent children and therefore the diagnosis may be difficult to make. We report tinea pedis in five children presenting as unilateral inflammatory lesions of the sole which was not readily diagnosed. The pathogen in all of our cases was Trichophyton rubrum. PMID- 10469407 TI - The natural history of primary herpes simplex type 1 gingivostomatitis in children. AB - Herpetic gingivostomatitis is the most common specific clinical manifestation of primary herpes simplex infection in childhood. The aim of the present study was to describe the clinical signs, symptoms, viral shedding, serologic findings, and complications in community-acquired gingivostomatitis. We prospectively followed children with herpes simplex type 1 gingivostomatitis lasting less than 72 hours. Clinical examination and viral culture were repeated every 2 to 3 days as long as symptoms or signs persisted. Thirty-six children (ages 12-77 months) were included in the study. Mean duration of oral lesions was 12.0+/-3.4 days; extraoral lesions (in 26 children), 12.0 +/-3.9 days; fever, 4.4+/-2.4 days; and eating/drinking difficulties, 9.1+/-3.0 and 7.1+/-3.1 days, respectively. In all children, viral cultures of the oral lesions were positive for herpes simplex virus (HSV) type 1; viral shedding persisted for a mean of 7.1+/-2.5 days (range 2-12 days). The main complications were dehydration, with three children hospitalized for intravenous rehydration, and one case of secondary bacteremia. Herpetic gingivostomatitis is a relatively severe manifestation of primary HSV type 1 infection in young children. PMID- 10469408 TI - A study of cutaneous tuberculosis in children. AB - Sixty-three children out of a total of 199 patients seen with cutaneous tuberculosis during a 7-year period were included in this study. Culture was positive in only four, and the diagnosis was based on clinical examination, tuberculin reaction, histopathology, and response to antitubercular therapy. Forty had lupus vulgaris (LV) and 23 scrofuloderma (SD). The lower half of the body was predominantly affected in those with LV, and keratotic and hypertrophic forms were frequently encountered. LV planus mainly affected the face. Ulcerative and atrophic types of LV were infrequent. Extensive lesions in three children led to disfiguring scars and contractures. Scrofuloderma often involved the cervical group of lymph nodes followed by the inguinal, submandibular, and axillary groups. As compared to skin tuberculosis in adults, regional lymph node involvement in LV was more common, and a combination of both LV and SD was less frequent in children. No difference in clinical presentation could be detected between the BCG vaccinated and unvaccinated children. Tuberculous infection either in the lungs or the bones was present in eight children. An HIV test done in five patients with widespread lesions was negative. Irregular therapy or late diagnosis leading to serious complications, inadequate parental or community support, and lack of awareness among practitioners are the problems to be remedied. PMID- 10469409 TI - Panniculitis in juvenile dermatomyositis. AB - Panniculitis is a rarely reported clinical finding in dermatomyositis. We present a 14-year-old African American boy with a 4-year history of dermatomyositis referred for evaluation of tender, indurated plaques and nodules on the trunk and proximal extremities. A biopsy specimen revealed epidermal and dermal changes consistent with dermatomyositis. Although calcification was absent, a striking lobular panniculitis was observed. A total of seven cases of clinical panniculitis in association with dermatomyositis have been published. As in our patient, it typically presents as indurated, tender plaques and nodules on the arms, thighs, and buttocks. Although in the vast majority of cases panniculitis found in association with dermatomyositis is subclinical and represents an incidental histopathologic finding, the case we present further emphasizes the need to recognize panniculitis as a rare, but clinically relevant pathologic feature seen in certain lesions of juvenile dermatomyositis. PMID- 10469410 TI - Generalized lupus panniculitis and antiphospholipid syndrome in a patient without complement deficiency. AB - Generalized chronic cutaneous lupus including lupus panniculitis in childhood is rare and usually occurs in the setting of genetic complement deficiencies. The association with antiphospholipid syndrome is even more rare. We report a 13-year old girl with extensive lupus panniculitis since the age of 8 months and no evidence of complement deficiency. She recently developed antiphospholipid syndrome characterized by anticardiolipin antibodies and digital necrosis. PMID- 10469411 TI - Wegener granulomatosis in a child: cutaneous findings as the presenting signs. AB - Wegener granulomatosis (WG) is a systemic disease that is particularly unusual in children. A limited form has been described without renal involvement. We report a 14-year-old girl in whom the disease started with acneiform nodular and papular lesions on the forehead. Later necrotic ulcers developed on her forehead, arms, and buttocks. The cutaneous lesions were associated with upper and lower respiratory tract involvement, low-grade fever and arthralgias. Subsequently clinical and laboratory evaluations (increased ESR; leukocytosis and presence of serum IgG antibodies cANCA = 1:160), with chest roentgenograms revealing pulmonary densities and parenchymal infiltration, suggested the diagnosis of WG. The histologic findings of a cutaneous biopsy specimen were ulceration of the epidermis with diffuse neutrophilic inflammatory infiltrate and a late-stage small vessel vasculitis in the dermis. Histopathology of the nasal mucosa was characterized by a granulomatous process with a dense lymphohistiocytic infiltrate with few giant cells, a finding that confirmed the diagnosis of WG. No renal involvement was present. One month of cyclophosphamide (125 mg/day) and prednisone (70 mg/day) therapy markedly improved the patient's clinical condition. At present, 1 year later, she is free from any signs of the disease. According to the literature, the frequency of cutaneous lesions in WG ranges from 16% to 46%. They are the presenting sign only in 6% of patients. Cutaneous lesions are even more uncommon in children. In particular, an "acneiform" presentation is a rare finding in WG. PMID- 10469412 TI - Familial multiple basaloid follicular hamartomas: A report of two affected sisters. AB - Basaloid follicular hamartoma (BFH) is one of several benign skin tumors that may occur in multiple and solitary forms. While the histologic findings of BFH may be observed in a variety of clinical settings, familial multiple BFH represents a distinct clinical entity characterized by tiny flesh-colored papules scattered predominantly over the face. We present two sisters with this rare condition and review the clinical and histologic differential diagnosis of familial multiple BFH. PMID- 10469413 TI - Bronchogenic cyst presenting as a nodular lesion. AB - We report a subcutaneous bronchogenic cyst in a 7-year-old girl with unusual presentation as a nodular lesion. The cyst was excised and no connection with deeper structures was found. Histologic examination showed the typical features of a bronchogenic cyst. The characteristic clinical and histologic features of bronchogenic cysts are reviewed. The problems of differential diagnosis are also discussed. PMID- 10469414 TI - Urticarial eruption associated with rheumatic fever in a child. AB - The classic description of erythema marginatum is of a few asymptomatic erythematous patches or plaques with a characteristic central clearing which may coalesce and develop a polycyclic configuration. We report a boy with a urticarial eruption without the classic lesions of erythema marginatum in whom the diagnosis of rheumatic fever was subsequently made. Histology of the urticarial lesions revealed a mixed, superficial, perivascular infiltrate with prominent eosinophils. Rheumatic fever should be considered in children with fever and urticaria-like eruptions even in the absence of the classically described erythema marginatum. PMID- 10469415 TI - Prepubertal hidradenitis suppurativa: two case reports and review of the literature. AB - Hidradenitis suppurativa (HS) is a chronic suppurative scarring disease of apocrine sweat gland-bearing skin in the axillary, anogenital, and, rarely, the breast and scalp regions. Females are more commonly affected than males and it is usually seen at puberty or later. We report two girls with prepubertal hidradenitis suppurativa whose initial presentation predated any signs of puberty. This early onset is very rare and its etiology remains unknown. Severe disease can be seen in prepubertal children and surgical intervention is effective in these cases. PMID- 10469416 TI - Monilethrix--improvement by hormonal influences? AB - Monilethrix is a hereditary hair disorder that occurs monosymptomatically or as a monilethrix syndrome combined with other ectodermal anomalies. We report two siblings with the triple combination of monilethrix, keratosis pilaris, and koilonychia. In one of our patients, the hair disorder improved, with maximum hair growth up to 8 cm in length after her first menstrual period occurred, suggesting that hormonal influences may improve the hair disorders in monilethrix. PMID- 10469417 TI - A sporadic case of congenital hypotrichosis simplex of the scalp: difficulties in diagnosis and classification. AB - Hereditary hypotrichosis simplex of the scalp is a genotrichosis characterized by a hair defect limited to the scalp in the absence of other ectodermal or systemic abnormalities. Only large pedigrees consistent with autosomal dominant transmission have been described to date. In this article the clinical and scanning electron microscopy findings of a nonfamilial case of congenital scalp hypotrichosis simplex are reported. In some patients the diagnosis of sporadic hypotrichosis simplex of the scalp should be considered after ruling out all other possible causes of congenital and hereditary hypotrichosis. PMID- 10469418 TI - Alopecia universalis in a patient with common variable immunodeficiency. AB - A 12-year-old boy with common variable immunodeficiency (CVI) who developed severe alopecia is presented. His sister also had alopecia and recurrent infections and died of lung infection at the age of 7 years. The loss of hair in both children was total; the pathology of a scalp skin biopsy specimen was typical for alopecia areata. The boy was subjected to clinical and immunologic evaluation and the results were compatible with common variable immunodeficiency. PMID- 10469419 TI - Multiple granular cell tumors and growth hormone deficiency in a child. AB - Granular cell tumor is an uncommon benign tumor occurring on the skin as a single nodule. Multiple tumors are very rare, particularly in children. We describe a child with multiple granular cell tumors on the skin in association with growth hormone deficiency. The occurrence of multiple granular cell tumors in association with other clinical manifestations in childhood is discussed. PMID- 10469420 TI - Depilation in a 6-month-Old with hypertrichosis: A case report. AB - Hypertrichosis in the pediatric age group can be troubling to both patients and parents. There is no well-established method for managing this problem in young children. We describe the successful use of a cream depilatory agent for removal of excess hair from the face and body of a 6-month-old girl. Excellent cosmetic results were obtained. The risks and benefits of the use of depilatory cream in young patients are analyzed. Other options for hair removal in children are also reviewed. PMID- 10469421 TI - Effect of penicillin G on corium thickness in linear morphea of childhood: An analysis using ultrasound technique. AB - Linear morphea is the most common form of localized scleroderma in children. The pathogenesis of this disorder is unknown. We report a child with a 6-year history of linear morphea in whom intravenous administration of 5 MU aqueous penicillin G three times a day for 10 consecutive days caused reduction in the thickness of the corium as demonstrated by ultrasound measurements. PMID- 10469422 TI - Calcipotriol in vitiligo: a preliminary study. AB - A large variety of therapeutic agents have been tried for the treatment of vitiligo, but psoralens continue to be the main treatment. Twenty-one patients age 5 to 17 years with vitiligo were enrolled in this study. The children were advised to apply calcipotriol 50 microg/g in the evening and expose themselves to sunlight the next day for 10 to 15 minutes. The patients were followed at 3-week intervals. Initial repigmentation occurred in the majority of children after 6 to 12 weeks of treatment. Marked to complete repigmentation was seen in 10 of 18 patients. Four patients showed moderate improvement while the remaining four patients showed minimal or no improvement. No patient developed new lesions. The repigmentation was cosmetically excellent in the majority of children. All patients tolerated the calcipotriol well except for three patients who complained of mild irritation on application. All of the laboratory investigations, including serum calcium levels remained normal. The rationale for this study originated from recent advances in the understanding of vitiligo at the molecular level. Furthermore, development of hyperpigmentation in patients with psoriasis receiving treatment with PUVA and calcipotriol has been observed. Our results are encouraging and offer a new and potentially efficacious treatment for this pigmentation disorder in children. PMID- 10469423 TI - A pilot study to assess the safety and efficacy of topical calcipotriol treatment in childhood psoriasis. AB - Childhood psoriasis is more extensive and severe compared with that of adults. Therefore an effective and safe treatment modality is needed. Although a few studies of childhood psoriasis indicate that treatment with calcipotriol is safe and effective, short-term studies cannot reflect the exact effect of calcipotriol on systemic calcium homeostasis. Our purpose was to study the long-term efficacy and safety of calcipotriol for childhood psoriasis. An uncontrolled pilot study, with long-term follow-up for as long as 106 weeks, using open-label calcipotriol ointment was conducted in 12 psoriasis patients less than 15 years of age. Response to treatment was assessed by the psoriasis area and severity index (PASI) and serum 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 levels, as well as routine laboratory analyses including serum calcium and phosphate, which were measured before and after the course of treatment. At the end of the study, the patients showed significant improvement in PASI scores compared with the baseline level. No serious side effects, including those related to calcium homeostasis, were detected. The mean values of 1,25-dihydroxyvitamin D3, however, were decreased and half of the patients had levels below the normal range. In conclusion, it is thought that calcipotriol ointment is an effective treatment modality for long-term use in childhood psoriasis. However, although not lowering serum calcium and phosphate levels, the long-term use of calcipotriol in childhood may possibly decrease the serum values of endogenous vitamin D. Therefore monitoring of vitamin D metabolites may be necessary during calcipotriol therapy. More investigative studies are needed to resolve this issue. PMID- 10469424 TI - Infiltrated lesions on a child's face. PMID- 10469425 TI - What syndrome is this? Oro-facio-digital type 1 syndrome of Papillon-Leage and Psaume. PMID- 10469426 TI - Vascular lesion laser: practical techniques or some "light" suggestions. PMID- 10469427 TI - Successful treatment of eosinophilic pustulosis with oral cimetidine. PMID- 10469428 TI - Atrophy of myocardium and its myocytes by left ventricular assist device. PMID- 10469429 TI - Different cell types within the sinoatrial node. PMID- 10469430 TI - Epidemiology of neurofibromatosis type 1. AB - The prevalence of neurofibromatosis type 1 (NF1) is about 1/3,000. There are no known ethnic groups in which NF1 does not occur or is unusually common. The prevalence is somewhat higher in young children than in adults, a difference that probably results at least in part from the early death of some NF1 patients. NF1 is fully penetrant in adults, but many disease features increase in frequency or severity with age. The reproductive fitness of NF1 patients is reduced by about one-half. About half of all cases result from new mutations. The estimated rate of new NF1 mutations is unusually high, but the basis for this high mutation rate is not known. Am. J. Med. Genet. (Semin. Med. Genet.) 89:1-6, 1999. PMID- 10469431 TI - Neurofibromatosis type 1: A model condition for the study of the molecular basis of variable expressivity in human disorders. AB - Neurofibromatosis type 1 (NF1) is a pleiotropic autosomal dominant disorder with marked variability of clinical expression. As in other heritable disorders, the mapping and cloning of the gene responsible for NF1 have increased our understanding of the pathogenesis of the condition. In particular, the phenotypic variability and variable expressivity can be studied using molecular techniques. In this article we summarize the current knowledge of genotype/phenotype correlation in NF1 and examine the potential molecular basis for variable expressivity. Am. J. Med. Genet. (Semin. Med. Genet.) 89:7-13, 1999. PMID- 10469432 TI - Hyperactive Ras as a therapeutic target in neurofibromatosis type 1. AB - The NF1 gene encodes neurofibromin, a GTPase-activating protein (GAP) for members of the p21(ras) (Ras) family, which negatively regulates Ras output by accelerating the conversion of active Ras. GTP to inactive Ras.GDP. Analysis of tumors from patients with neurofibromatosis type 1 (NF1) has shown biochemical evidence of hyperactive Ras as well as frequent loss of the normal NF1 allele, consistent with its role as a tumor suppressor gene. Taken together, these data suggest that novel therapeutics directed against components of the Ras signaling cascade might provide effective treatments for certain pathological complications of NF1. Here we summarize data that support a role for hyperactive Ras in NF1 disease, including Ras processing, activation, and down-regulation. We review targets for rational drug design, provide preliminary results, and discuss implications for future studies. Am. J. Med. Genet. (Semin. Med. Genet.) 89:14 22, 1999. PMID- 10469433 TI - Pathology of tumors of the peripheral nerve sheath in type 1 neurofibromatosis. AB - The two main peripheral nerve sheath tumors found in patients with neurofibromatosis, type 1 (NF1), are neurofibroma, a benign tumor, and malignant peripheral nerve sheath tumor (MPNST). The tumors are related in that most MPNSTs are thought to arise by malignant transformation of neurofibromas. Such an event occurs in about 2% of NF1 patients. There are five forms of neurofibroma; three of them-localized cutaneous neurofibroma when multiple, plexiform neurofibroma, and massive soft-tissue neurofibroma-are highly specific for NF1. Only two forms of neurofibroma, plexiform and localized intraneural neurofibroma, are significant precursors of MPNST. Massive soft-tissue neurofibromas are worrisome in that they may mask MPNST arising from one of the mentioned neurofibromas. The vast majority of MPNSTs are high-grade malignant tumors with a high rate of distant metastasis. The overall 5-year survival rate for patients with MPNSTs ranges from 34% to 52%. MPNSTs generally are solitary, deep-seated globoid or fusiform tumors. They are firm, fleshy, tan, and often focally to extensively necrotic, and they invade surrounding soft tissue. On histological examination, MPNSTs are most often hypercellular, hyperchromatic, fasciculated, and mitotically active tumors. Low-grade tumors account for only about 10-15% of cases. Twenty percent of MPNSTs have unusual and potentially misleading histological features, such as epithelioid cells and divergent mesenchymal or glandular differentiation. Am. J. Med. Genet. (Semin. Med. Genet.) 89:23-30, 1999. PMID- 10469434 TI - Plexiform neurofibromas. AB - Plexiform neurofibromas are among the most common and debilitating complications of neurofibromatosis type 1 (NF1). They account for substantial morbidity, including disfigurement, functional impairment, and may even be life threatening. Plexiform neurofibromas are also subject to transformation into malignant peripheral nerve sheath tumor (MPNST), a complication that is refractory to treatment both because of a paucity of effective therapies for malignant soft tissue sarcomas in general, and because of the delay in diagnosis that results from change of a small portion of a large pre-existing tumor. The current mainstay of treatment of plexiform neurofibromas, and of MPNST for that matter, is surgical resection. The major variables are the timing and means of identification of plexiform neurofibromas, methods of follow-up, and indications for surgery. There is no established means of medical treatment, but research into the molecular pathogenesis of NF1, as well as advances in tumor therapy in general, are opening the way towards clinical trials for plexiform neurofibroma. Am. J. Med. Genet. (Semin. Med. Genet.) 89:31-37, 1999. PMID- 10469435 TI - Intracranial gliomas in neurofibromatosis type 1. AB - Optic pathway gliomas and brainstem gliomas are the predominant intracranial neoplasms associated with neurofibromatosis type 1 (NF1). Before the past 15 years, studies of optic pathway gliomas in NF1 were hampered by the inaccurate diagnosis of NF1, the unavailability of noninvasive neuroimaging techniques, and the frequent rendering of what would now be considered unnecessary, overly aggressive therapy. When studied systematically, these tumors behave in a much more benign fashion than their counterparts in children who do not have NF1. While they may cause symptoms in as many of 50% of cases, progression to the point where specific intervention is deemed necessary is unusual. Consequently, screening neuroimaging of asymptomatic patients is unwarranted. Because optic pathway tumors universally arise in children younger than 7 years of age, all such children should undergo yearly ophthalmologic evaluations and annual assessments of growth to monitor for signs of precocious puberty. Am. J. Med. Genet. (Semin. Med. Genet.) 89:38-44, 1999. PMID- 10469436 TI - Cognitive impairment in neurofibromatosis type 1. AB - Neurofibromatosis type 1 (NF1) is associated with a broad range of relatively nonspecific cognitive impairments, including low IQ, learning disabilities, and behavioral difficulties. While early studies indicated that the cognitive phenotype of NF1 resembles that of nonverbal learning disabilities (NLD), later research has found that the impairments are broader and do not fit the NLD profile well. Language-based deficits and executive dysfunction have also been found in empirical studies. There is some evidence that neuropsychological impairment may be correlated with the underlying central nervous system involvement of NF1, but this theory has not been consistently supported across studies. Further work clarifying the cognitive phenotype of NF1 is needed, especially investigations that employ appropriate comparison groups matched for intellectual level. Am. J. Med. Genet. (Semin. Med. Genet.) 89:45-52, 1999. PMID- 10469437 TI - The beginning of any great matter PMID- 10469438 TI - Neurofibromatosis 1 PMID- 10469439 TI - Neuronal survival using a resorbable synthetic conduit as an alternative to primary nerve repair. AB - Clinically optimal situations for primary nerve repair are rarely observed. Crushed nerve ends result in either suboptimal repair or a need for nerve grafting. Functional results after nerve surgery are relatively poor, including major sensory deficits, which may be due to the death of primary sensory neurons that follows the nerve injury. The aim of this study was to determine if using polyhydroxybutyrate (PHB), a resorbable nerve conduit, could be an alternative to primary nerve repair in reducing loss of neurons. The superficial radial nerves in 20 cats were sectioned bilaterally and primarily repaired microsurgically by the use of two different strategies; either wrapping the nerve ends in sheets of PHB or epineurally suturing of the nerve. After 6 or 12 months, the surviving neurons within the dorsal root ganglia [C5-T1] were counted. No statistically significant differences were found between the two methods. This may imply a future possibility of using PHB as a synthetic nerve graft in situations where suboptimal primary repair or nerve grafts are the alternatives. PMID- 10469440 TI - Local recombinant tissue plasminogen activator (rt-PA) thrombolytic therapy in microvascular surgery. AB - Vascular thrombosis remains a dreaded complication of any microvascular procedure, be it composite tissue transfer or replantation of amputated limbs or parts. Despite the tremendous advances in microvascular-related technologies and the accumulated surgical skills, failures caused by occlusion of anastomosed vessels remain a continuous source of frustration to all microsurgeons alike. Several anticoagulation and antiplatelet protocols have been proposed to be used in conjunction with microvascular surgery. More recently, thrombolytic drugs such as urokinase, streptokinase, and thrombolysin have been introduced, yet their systemic effect on hemostasis remains an undesirable side effect. We present our experience with local intra-arterial, intravenous, and soft-tissue injection of recombinant tissue plasminogen activator rt-PA in replantation surgery in three consecutive patients. Arterial thrombi are managed by intra-arterial rt-PA infusion with the catheter placed proximal to the arterial anastomosis. Venous thrombi are best lysed by infusing rt-PA in an engorged vein of the replanted limb. In replanted digits, direct intravenous infusion is not possible. In such situations, injection of rt-PA in the pulp soft tissues may result in successful salvage. We believe this agent also has a role in microvascular composite tissue transfer in preventing free flap failures as well as in salvaging failing flaps. PMID- 10469441 TI - Modified technique for kidney transplantation in mice. AB - This study describes a new method for joining the donor ureter to the recipient bladder during mouse kidney transplantation. The donor left kidney was harvested using methods previously published, except that bladder tissue was not harvested with the end of the ureter. The recipient left kidney was removed and the donor kidney was attached using end-to-side anastomosis. The recipient bladder was pierced with a 21-gauge needle allowing curved forceps to be inserted through the bladder, to pull through the ureter, and the periuretal tissue was stitched to the exterior wall of the bladder. The donor ureter was allowed to retract inside the bladder. Following a right nephrectomy, grafts were monitored by blood serum creatinine and urea. With a technical success rate of 83%, this technique reduced donor harvest time by 20 minutes and ureter attachment time by 15 minutes making it the best method available for mouse kidney transplantation. PMID- 10469442 TI - Local administration of neurotrophic growth factor in subcutaneous silicon chambers enhances the regeneration of the sensory component of the rat sciatic nerve. AB - An experimental model for local administration of neurotrophic growth factor (NGF) in peripheral nerve lesions is tested. The model consists of a subcutaneous reservoir connected to the sciatic nerve neurorrhaphy. The right sciatic nerves were exposed, severed, and repaired at a level 1.5 cm proximal to their trifurcation. Then, a dome-shaped silicone reservoir connected to the proximal end of a silicone tube was placed subcutaneously in the dorsum of the experimental animal. The distal end of the connecting tube was located in the nerve neurorrhaphy. Two experimental groups were made: Group A (n = 90) received daily doses of a solution containing NGF-7S during the first 4 weeks after surgery and a single weekly dose thereafter. Within this group, three subgroups of 30 rats each were made: A-4 sacrificed 4 weeks after surgery, A-8 sacrificed after 8 weeks, and A-12 after 12 weeks. Group B (n = 90) received the same vehicle solution without NGF under the same schedule and volume as in Group A. Three subgroups were also made as in Group A depending on the survival period. In order to locate the neurons in the dorsal root ganglia, the retrograde tracer horseradish peroxidase was administered at the proximal stump of the sciatic nerve (tibialis branch), which was severed 1 cm distal to the sciatic trifurcation. In respect of the nonoperated side, the percentage between the number of dorsal root ganglia neurons in the NGF-treated group was significantly higher than in the control group (P < 0.001). These results demonstrate that percutaneous administration of multiple doses of NGF in this model enhances sensory nerve regeneration after sciatic lesions evaluated by horseradish peroxidase labeling of dorsal root ganglia neurons. PMID- 10469443 TI - Evaluation of collateral sprouting after end-to-side nerve coaptation using a fluorescent double-labeling technique. AB - The mechanism of end-to-side neurorrhaphy is believed to be by collateral sprouting, although evidence for this is lacking. This study validates whether axonal sprouting originates from the donor intact nerve by collateral sprouting with the use of a fluorescent double-labeling technique. End-to-side neurorrhaphy was performed on adult female Sprague-Dawley rats. Eight and 12 months postoperatively, animals were injected with true blue and diamidino yellow into the tibialis anterior and/or gastrocnemius muscles and were transcardially perfused with fixative after 7 days of retrograde transport. The lumbar enlargement and the dorsal root ganglia from L3 to L6 were harvested and serial sectioning and fluorescent microscopy were performed. No double-labeling neurons were observed in control animals, but a group of neurons that were greenish or yellowish in color were seen with single labeling. Double-labeling neurons, however, were seen in animals treated with end-to-side neurorrhaphy whether or not perineurotomy was performed. These results demonstrate that one parent nerve fiber can emanate another axon by collateral sprouting following end-to-side neurorrhaphy. We hypothesize that the causes of collateral sprouting might result from "switching signals" and/or "switching factors." PMID- 10469444 TI - Sensory abnormalities after the seventh cervical nerve root transfer. AB - Twenty-four patients with root avulsion of brachial plexus were treated with the contralateral C7 root transfer. The area of sensory abnormalities was found on the thumb in 20 cases (83.33%), index finger in 14 cases (58.33%), middle finger in 8 cases (33.33%), ring finger in 3 cases (12.5%), little finger in 11 cases (45.83%), on the radial side of forearm in 9 cases (37.5%), thenar area in 12 cases (50%), and hypothenar area in 8 cases (33.33%). No patient had normal sensation after the seventh cervical root transfer. Sensory abnormalities disappeared within 3 months in all except one patient, who still had sensory abnormalities on the tip of index finger for more than 20 months after the operation. PMID- 10469445 TI - Neovascularized bone grafts: experimental investigation. AB - Vascularized bone grafts are standardized procedures in reconstructive surgery but there are some disadvantages: donor site morbidity, limited number of "natural" donor sites, and complex technique. In this study, we test the possibility of creating a "neovascularized" bone graft utilizing a vascular implantation procedure in a rabbit model. Sixteen New Zealand adult white rabbits were used. In each animal, two iliac crest bone grafts (7 x 7 x 10 mm) were harvested. Vascular implantation of the right superficial femoral vessels was performed in one of the two grafts, which was wrapped in a silicone envelope to avoid neovascularization from the surrounding tissues and positioned in a subcutaneous pocket in the right medial thigh. On the left side, the bone block, wrapped in the silicone envelope, was buried subcutaneously without vascular implantation. The operated animals were divided into two groups: Group I included eight rabbits explanted 4 weeks postoperatively and Group II included eight rabbits explanted 8 weeks postoperatively. Tetracycline injection was performed 72 hours preexplantation to evaluate new bone formation. Selective colloidal ink injection in the axial artery was performed to investigate the neovascularization before inclusion in poly-methyl-methacrylate (PMMA). Histological examination was performed in all explanted specimens comparatively. Histological examination 8 weeks after surgery showed a marked neovascularization, with normal bone cells. Tetracycline labeling showed new bone formation with a normal pattern. In all nonvascularized specimens, no viable cells or neovascularization and no bone formation were found. The vascular implantation procedure can induce a good neovascularization with new bone formation in a small bone graft. The possibility of neovascularization induction by the simple vascular implantation procedure has several clinical implications in reconstructive surgery. PMID- 10469446 TI - Revascularization of turnover sternum: A definitive treatment for intractable funnel chest. AB - Twelve intractable funnel chests in young adults were treated with revascularization of the turnover sternum. The sternum and costal composite tissue were resected at the outskirt of the depressed area. Special attention was paid to the dissection of the vascular pedicle at both sides of the internal mammary vessels. The recipient vessels at one side were left long, so were the donor vessels at the other side. A segment of the rib at the exit of the recipient internal mammary vessels had to be removed to accommodate the vessels and to facilitate vascular anastomosis. Vascular anastomosis was accomplished with loupes (Keeler, sixfold magnification) in five patients because the direction of the vessels is vertically oriented. In four cases, artery and vein grafts were taken from the other side of the internal mammary vessels not bound for vascular anastomosis for length discrepancy of the vessels, while the remaining cases had direct vascular anastomosis without vascular grafting. Revascularization of the turnover sternum was performed successfully without vascular compromise. The patients all recovered well with much improved physical condition. Only one patient sustained spontaneous pneumothorax 1 month after the operation. Postoperative three-dimensional computed tomographic (CT) scan revealed increment of thoracic cage volume for 9-17%. Follow-up CT scan 2 years later revealed even more improved thoracic cage expansion. PMID- 10469447 TI - Nonrandom fusion of L-plastin(LCP1) and LAZ3(BCL6) genes by t(3;13)(q27;q14) chromosome translocation in two cases of B-cell non-Hodgkin lymphoma. AB - The LAZ3(BCL6) gene on chromosome band 3q27 is nonrandomly disrupted in B-cell non-Hodgkin lymphoma (B-NHL) by chromosomal translocations clustered within a 3.3 kb MTC (major translocation cluster) located between the two first noncoding exons. These translocations generally result in the expression of a chimeric mRNA transcript between the LAZ3 gene and sequences derived from the partner chromosome. Using RACE RT-PCR, we previously demonstrated fusion of LAZ3 with the RhoH/TTF gene, a hemopoietic cell-specific small GTPase involved in cytoskeleton organization, and with the BOB1/OBF1 gene, a B-cell-specific coactivator of octamer-binding transcription factors, following translocations t(3;4)(q27;p13) and t(3;11)(q27;q23), respectively. Here we report the identification of the L Plastin(LCP1) gene as a novel LAZ3 partner in chimeric transcripts resulting from a t(3;13)(q27;q14) translocation, in two cases of B-cell lymphoma. As a consequence of the translocation, the 5' regulatory region of each gene was exchanged, creating both LCP1-LAZ3 and reciprocal LAZ3-LCP1 fusion transcripts in one case, and only a LCP1-LAZ3 fusion transcript in the other. The 13q14 chromosome region is frequently disrupted in various proliferative disorders, and the LCP1 gene defines a new breakpoint site in this region. This gene encodes an actin-binding protein and is the second LAZ3 partner gene, with the RhoH/TTF gene, involved in actin cytoskeleton organization. Genes Chromosomes Cancer 26:97 105, 1999. PMID- 10469448 TI - Regional reproducibility of microsatellite instability in sporadic colorectal cancer. AB - The reproducibility of microsatellite instability from different regions of the same sporadic colon cancer has not been addressed. We therefore microdissected and extracted DNA from three to nine separate regions of 13 highly unstable sporadic colon cancers. Each region was then evaluated by polymerase chain reaction amplification of 17 microsatellites: 10 tetranucleotide repeats, 2 noncoding mononucleotide repeats (BAT-26 and BAT-40), and 5 coding mononucleotide repeats (TGFBRII, BAX, MSH3, MSH6, IGFIIR). Microsatellite instability showed 100% regional reproducibility with respect to either the panel of 10 tetranucleotide repeats or BAT-26, and nearly 100% reproducibility with BAT-40, although regional variation in the percent instability and the size of unstable alleles was present. TGFBRII was more frequently mutated than any other coding mononucleotide repeat; frame shifts in this gene were identified in nearly every region of every tumor. Each of the five coding repeats showed regional variability in at least one tumor, and 10 of the 13 tumors showed variability with at least one coding repeat. This variability took the form of different mutant alleles (TGFBRII, BAX, MSH3) or mutations present in some but not all regions of a tumor (MSH6, IGFIIR, BAX, MSH3). We conclude that the regional reproducibility of generalized microsatellite instability as measured by noncoding repeats indicates that sampling is not a problem in these highly unstable tumors, and that the mismatch repair deficiency phenotype is acquired in the very late adenoma stage or early cancer stage of sporadic colonic tumorigenesis. The high frequency of TGFBRII mutations is consistent with acquisition of these mutations at a similar stage of tumorigenesis. The regional variability with respect to the presence or absence of a mutation in the other four coding mononucleotide repeats could lead to sampling error and is consistent with a somewhat later time of acquisition of these mutations. Genes Chromosomes Cancer 26:106-114, 1999. PMID- 10469449 TI - A novel chromosomal region of allelic loss, 4q32-q34, in human osteosarcomas revealed by representational difference analysis. AB - Representational difference analysis (RDA) of a human osteosarcoma xenograft resulted in the isolation of four tumor-associated homozygously deleted DNA fragments, all originating from chromosome 4, region q32-q34. Southern blot analysis using the RDA fragments and interphase FISH analysis using PACs corresponding to these RDA fragments revealed allelic loss of the 4q32-q34 region in 17 of 27 (63%) osteosarcomas tested. These results suggest the involvement of tumor suppressor gene(s) within this chromosomal region in osteosarcoma development. The RDA fragments and corresponding PAC clones will be instrumental in the isolation of such gene(s). Genes Chromosomes Cancer 26:115-124, 1999. PMID- 10469450 TI - A high frequency of tumors with rearrangements of genes of the HMGI(Y) family in a series of 191 pulmonary chondroid hamartomas. AB - Pulmonary chondroid hamartomas (PCHs) are benign mesenchymal tumors that often are characterized by specific chromosomal aberrations. Herein we report our cytogenetic and molecular cytogenetic (FISH) studies on 191 PCHs, including 48 previously published cases. In this series, 134/191 PCHs (70.2%) showed either abnormalities of chromosomal bands 6p21 (21 tumors), 12q14-15 (95 tumors), or had other abnormalities (18 tumors). Two tumors had a 6p21 aberration together with a 12q14-15 aberration. The most frequent translocations were t(12;14)(q15;q24) (19 cases) and t(6;14)(p21. 3;q24) (18 cases), both in either simple or complex form. By FISH with cosmids spanning the gene encoding the high-mobility-group protein HMGIC, we were able to show a rearrangement within or close to HMGIC in all tumors with 12q14-15 abnormalities tested, in 11 tumors with an apparently normal karyotype, and in 4 tumors with complex abnormalities without cytogenetically visible alterations of chromosomes 12. Rearrangements of HMGIY or its immediate surroundings were shown for 21 cases with 6p21 aberrations and three cases with other chromosomal abnormalities but without cytogenetically visible alterations of chromosomes 6. Genes Chromosomes Cancer 26:125-133, 1999. PMID- 10469451 TI - Allelic loss at 1p34, 13q12, 17p13.3, and 17q21.1 correlates with poor postoperative prognosis in breast cancer. AB - Allelic losses of tumor suppressor genes (TSGs), or the chromosomal regions harboring them, in tumor DNA may become useful postoperative prognostic indicators. To examine whether specific allelic losses might correlate with postoperative survival in a 5-year prospective follow-up, we tested tumors from a cohort of 264 breast cancer patients for allelic losses of 18 microsatellite markers representing either a known TSG or a region where genetic alterations are frequent in breast tumors. Patients whose tumors had lost an allele at 1p34, 13q12, 17p13.3, or 17q21.1 had significantly higher risks of postoperative mortality than those whose tumors retained both alleles at those loci (at 1p34, a 5-year mortality rate of 29% among patients with losses vs. 7% with retentions, P = 0. 0008; at 13q12, 31% vs. 10%, P = 0.0062; at 17p13.3, 24% vs. 13%, P = 0.026; and at 17q21.1, 31% vs. 13%, P = 0.0047). Furthermore, combined losses at 13q12 and 17p13.3 increased the predicted postoperative mortality risks by a factor of 9.6 (5-year mortality rate of 42% vs. 5% with retentions, P = 0.0001), and combined losses at 1p34 and 17p13.3 raised the predicted postoperative mortality risks by a factor of 8.6 (27% vs. 3%, P = 0.0064). We conclude that allelic losses at these loci can serve as negative prognostic indicators to guide postoperative management of patients. Genes Chromosomes Cancer 26:134-141, 1999. PMID- 10469452 TI - Characterization of topoisomerase II alpha gene amplification and deletion in breast cancer. AB - Topoisomerase IIalpha (TOP2A) is a key enzyme in DNA replication and a molecular target for many important anticancer drugs. TOP2A is amplified or deleted together with amplification of the closely located ERBB2/HER-2/neu oncogene in breast cancer. We characterized the copy number aberrations of TOP2A and ERBB2 in 136 primary breast tumors by FISH. Among the 70 primary tumors with ERBB2 amplification, amplification of TOP2A was found in 29 (41%); 30 tumors (43%) showed a physical deletion of TOP2A; and the copy number for TOP2A was not altered in 11 tumors with ERBB2 amplification (16%). No TOP2A gene aberrations were identified in 65 primary tumors without ERBB2 amplification. Fiber FISH revealed that simultaneously amplified ERBB2 and TOP2A were not present in the same amplicon, because repetitive tandem repeat-like signals of ERBB2 and TOP2A were in separate DNA fibers. The deletion of TOP2A (seen in the MDA-361 cell line and in 31 primary tumors) was interstitial, spanning less than two megabases of DNA. Mean copy numbers of TOP2A (2.4 +/- 0.6 for TOP2A vs. 4.9 +/- 1.1 for chromosome 17 centromere) suggest that the deletion of TOP2A occurs before polyploidization of the genome. Eight primary tumors with high-level ERBB2 amplification showed a new type of intratumoral heterogeneity; two different cell clones with either high-level amplification or deletion of TOP2A were found adjacent to each other in the same tumor. These results indicate that amplification of the ERBB2 oncogene is followed by complex secondary genetic aberrations, which lead to amplification or deletion of the TOP2A gene in a majority of tumors. Genes Chromosomes Cancer 26:142-150, 1999. PMID- 10469453 TI - Chromosome band 9p21 is frequently altered in malignant peripheral nerve sheath tumors: studies of CDKN2A and other genes of the pRB pathway. AB - Malignant peripheral nerve sheath tumors (MPNSTs) are frequently associated with the disease neurofibromatosis type 1. Only few recurrent cytogenetic changes have been reported, including rearrangements of the short arm of chromosome 9. By fluorescence in situ hybridization with a centromere 9 probe, and by allelic imbalance studies with seven 9p21-23 markers in nine familial and three sporadic MPNSTs, we found interstitial deletions that supported CDKN2A as a possible target gene. Nine MPNSTs showed aberrations of CDKN2A by Southern blot analyses, and in four of these, expression of CDKN2A could not be detected by Northern blot analysis. No mutations of CDKN2A were identified by sequencing of the coding region, and gene inactivation by promoter methylation was not found. In the 9p allelic imbalance studies, a novel allele was detected at one locus in one tumor. Analyses of additional markers (n = 8) excluded mismatch repair deficiency as an important mechanism in the genesis of these tumors. The tumors were analyzed further for alterations in other candidate cell cycle-associated genes. In total, 11/12 MPNSTs showed DNA changes in one or more of the genes CDKN2A, CDKN2B, RB1, CDK4, MDM2, and CCND2. The present study suggests that disruption of the pRB pathway is common in MPNST, and that dose reduction of CDKN2A is particularly frequent and contributes to MPNST development. Genes Chromosomes Cancer 26:151 160, 1999. PMID- 10469454 TI - A case of inv(8)(p11q24) associated with acute myeloid leukemia involves the MOZ and CBP genes in a masked t(8;16). AB - We report on a novel chromosomal aberration, inv(8)(p11q24), in an M5 acute myeloid leukemia. We show by fluorescence in situ hybridization and Southern blot analyses that a t(8;16)(p11;p13) is masked by this inversion. The translocation targets the MOZ gene from the 8p11 and the CBP gene from the 16p13 chromosomal regions. The breakpoints occur in the MOZ region encoding the acidic domain and in the 5' end of the CBP gene. These results provide further evidence for the multiple contribution of both MOZ and CBP genes in acute leukemias. Genes Chromosomes Cancer 26:161-165, 1999. PMID- 10469455 TI - Amplification of the 11q23 region in acute myeloid leukemia. AB - Cytogenetic abnormalities involving the 11q23 region are found in both acute lymphoblastic leukemia (ALL) and myeloid leukemia (AML). Molecular consequences of 11q23 translocations are the formation of chimeric genes, all of them involving the MLL (mixed-lineage leukemia) gene. To evaluate the usefulness of fluorescence in situ hybridization (FISH) in detecting MLL rearrangements in AML, we analyzed 181 patients with an MLL-specific probe. Among them, we detected three patients with multiple FISH signals, reflecting genomic amplification of this chromosomal region. Extra copies of MLL have been reported previously in four patients, but did not correspond to a true gene amplification. For the first time, we describe genomic amplification of the 11q23 region (up to more than 50 copies) in AML patients. This genomic amplification could affect MLL, but other genes in the vicinity could also be the primary target. Genes Chromosomes Cancer 26:166-170, 1999. PMID- 10469456 TI - Homozygous deletion at the 9q32-33 candidate tumor suppressor locus in primary human bladder cancer. AB - Loss of heterozygosity (LOH) on chromosome arm 9q is the most frequent genetic alteration found in superficial and invasive transitional cell carcinoma (TCC) in a previous microsatellite-based deletion mapping study of the bladder and upper urinary tract, indicating the presence of one or more important tumor suppressor genes (TSGs). One of the putative tumor suppressor loci on 9q (DBC1) was mapped to 9q32-33 and the candidate region was localized within a single YAC. We report here a case of superficial papillary TCC, which showed a homozygous deletion encompassing this candidate tumor suppressor region. The region of homozygous deletion spanned the interval between D9S275 and AFMA239XA9 at 9q32-33, and was estimated to be A (35), G-->T (35), G-->C (37), G-->T (38), C- >A (181), A-->T (182) and A-->G (182) in the low dose tumors were 5, 2, 11, 74, 0, 7 and 2%, respectively, and the distribution of mutations in tumors from animals treated with a high dose of BP were 3, 7, 13, 61, 15, 1 and 0%, respectively. Differences in the global mutation spectra (site and kind of all mutations) for the c-Ha-ras gene between the high and low dose group tumors were statistically significant (P < 0.004, Fisher's exact test) and the major difference between these two groups was C-->A (181) base substitutions. In summary, our data indicate that: (i) 79% of the BP/TPA skin tumors in CD-1 mice had c-Ha-ras mutations for the combined data for high dose and low dose tumors; (ii) the major mutations detected in BP/TPA skin tumors were G-->T transversions; (iii) the global mutation profile in the c-Ha-ras proto-oncogene in skin tumors obtained after initiation with a low dose of BP was different from that obtained after initiation with a high dose of BP. PMID- 10469613 TI - Reduced expression of the CDK inhibitor p27(KIP1) in rat two-stage bladder carcinogenesis and its association with expression profiles of p21(WAF1/Cip1) and p53. AB - The cyclin-dependent kinase (CDK) inhibitor p27(KIP1) exerts its growth suppressive effects by targeting the cyclin-CDK complexes. Reduced protein levels of p27(KIP1) have been reported in numerous human cancers and this has been attributed to increased degradation. However, few reports have addressed the significance of p27(KIP1) expression in chemical carcinogenesis of rodents. In a rat two-stage urinary bladder carcinogenesis model, with N-butyl-N-(4 hydroxybutyl)nitrosamine (BBN) initiation followed by promotion with sodium L ascorbate (Na-AsA), we evaluated the expression of p27(KIP1) protein using immunohistochemistry during various stages of urinary bladder carcinogenesis. In addition, we evaluated the mRNA expression profiles for p27(KIP1), p21(WAF1/Cip1) and p53 in tumors. Fisher 344 rats were initiated with 0.05% BBN in the drinking water for 4 weeks and then administered 5% Na-AsA in the diet. Immunohistochemical examination revealed p27(KIP1) protein to be constitutively expressed in normal urothelium, simple hyperplasia and in most papillary and nodular (PN) hyperplasias and small papillomas, but diminished or absent in large papillomas and in transitional cell carcinomas. An inverse correlation between expression of p27(KIP1) and cell proliferation was generally observed. Quantitation of mRNA by multiplex reverse transcription-PCR showed a significant downregulaton of p27(KIP1), p21(WAF1/Cip1) and p53 mRNA in tumors. More than 50% reduction in p27(KIP1) mRNA expression was observed in 42 and 47% of tumors at weeks 18 and 24, respectively; similar reduction in p21(WAF1/Cip1) mRNA expression was observed in 58 and 73% of tumors at weeks 18 and 24, and in p53 mRNA expression in 50 and 73% of tumors at weeks 18 and 24, respectively. None of the 25 tumors we examined by PCR-single-strand conformational polymorphism analysis had p53 mutations. These data imply that abnormal down-regulation of p27(KIP1), p21(WAF1/Cip1) and/or p53 in tumor cells may contribute to the malignant progression of tumors during rat two-stage bladder carcinogenesis. PMID- 10469614 TI - APC-dependent changes in expression of genes influencing polyamine metabolism, and consequences for gastrointestinal carcinogenesis, in the Min mouse. AB - The colorectal mucosa of pre-symptomatic individuals with familial adenomatous polyposis (FAP) contains elevated levels of the proliferation-associated polyamines. The Min mouse, like humans with FAP, expresses an abnormal genotype for the APC tumor suppressor gene. In order to determine how APC mutation influences intestinal tissue polyamine content, we measured steady-state RNA levels of ornithine decarboxylase (ODC), the first enzyme in polyamine synthesis, antizyme (AZ), a protein which negatively regulates ODC, and the spermidine/spermine N(1)-acetyltransferase (SSAT), the first enzyme in polyamine catabolism. RNA content was increased 6- to 8-fold in both the small intestine and colon for ODC, decreased significantly in the small intestine but not the colon for AZ and was not statistically different in either intestinal tissue for SSAT in Min mice compared with normal littermates. Consistent with the changes in ODC and AZ gene expression, small intestinal, but not colonic, polyamine content was elevated in Min mice compared with normal littermates. Treatment of Min mice with the specific ODC inhibitor difluoromethylornithine (DFMO) suppressed small intestinal, but not colonic, polyamine content and tumor number. These data indicate that small intestinal tissue polyamine content is elevated in Min mice by a mechanism involving APC-dependent changes in ODC and AZ RNA. Further, ODC enzyme activity, which is influenced by both ODC and AZ RNA levels and inhibited by DFMO, is consequential for small intestinal tumorigenesis in this model. In the FAP population, DFMO may be of value in the chemoprevention of small intestinal adenocarcinoma that remains a risk following colectomy. PMID- 10469615 TI - Interactions of apoptosis, proliferation and host age in the regression of the mouse mammary preneoplasia, TM3, carrying an unusual mutation in p53. AB - We have developed an in vivo model system of mouse mammary preneoplasias in order to examine the cell and molecular changes that occur during tumorigenesis. Most of these preneoplasias are characterized by an alveolar hyperplasia morphologically similar to that present in normal pregnant mammary gland, but have tumor forming capabilities ranging from very low to high. One of these hyperplasias, the TM3 HOG (transformed mammary hyperplastic outgrowth), forms tumors infrequently and has the unusual characteristic of spontaneous regression. We have observed that 7-8 months post-transplantation into the cleared mammary fat pad of a BALB/c mouse, the TM3 hyperplasia will begin to regress, leaving only a sparse ductal tree with remnant alveolar structures by 10-12 months post transplantation. We have sought to elucidate the mechanism of this regression by determining the apoptotic and proliferative rates of the alveolar cells during TM3 HOG development. Studies show that apoptotic rates in the TM3 HOG are consistently high (4-7%) at all times after transplantation. This apoptotic rate is higher than the rates found in other preneoplasias in our system and approach the rates observed in the normal involuting gland. An unusual p53 mutation, a serine insertion at codon 233, may be causally related to the high spontaneous apoptotic frequencies as well as elevated inducible apoptotic frequencies in TM3. In addition, a sudden decrease ( approximately 63%) in proliferation occurs around 8 months post-transplantation. Furthermore, transplantation experiments indicate that the ability of the 8-month-old host and/or mammary gland to support growth of preneoplastic mammary tissues is markedly diminished compared with 3- or 6-month-old hosts. The results presented here suggest that the persistent high apoptotic rates, concomitant with decreased proliferation rates, may be responsible for TM3's regression and implicate a unique mutant p53 as a causal factor. Additionally, the results suggest that host determinants can interact with specific molecular changes in the preneoplastic cells to influence growth and progression of the preneoplastic populations. PMID- 10469616 TI - Effect of energy restriction on tissue size regulation during chemically induced mammary carcinogenesis. AB - Energy restriction (ER) has documented beneficial effects on numerous diseases including cancer, yet the mechanism(s) that accounts for these effects is unknown. Experiments were designed to determine the effect of ER: (i) on the growth and development of the mammary gland; (ii) on the growth of carcinomas induced in the mammary gland by treatment with 1-methyl-1-nitrosourea (MNU); (iii) on rates of cell proliferation and apoptosis in pre-malignant and malignant mammary lesions. Mammary carcinogenesis was induced in female Sprague-Dawley rats by the i.p. administration of MNU (50 mg MNU/kg body wt) at 21 days of age. Rats were randomized to one of four dietary treatment groups: ad libitum fed or restriction of calorie intake to 90, 80 or 60% of ad libitum intake. ER reduced the ductal extension of the mammary gland into the fat pad in proportion to its effect on growth measured as body weight, however, the reduction in ductal branching, breast density and carcinoma volume by ER was greater than its effect on body weight. An animal's breast density was predictive of its carcinogenic response, irrespective of the level of ER imposed. While ER inhibited cell proliferation and induced apoptosis in pre-malignant and malignant mammary gland lesions, the magnitude of these effects make it unlikely that they fully account for the protective effects of ER against mammary carcinogenesis. PMID- 10469617 TI - Prostate cancer risk and polymorphism in 17 hydroxylase (CYP17) and steroid reductase (SRD5A2). AB - Prostate cancer is the most common malignancy in males and is the second most common cause of cancer mortality in American men. Polymorphisms have been identified in two genes, the 17-hydroxylase cytochrome P450 gene (CYP17) and the steroid 5-reductase type II gene (SRD5A2) that are involved with androgen biosynthesis and metabolism. The CYP17 A2 allele contains a T-->C transition in the 5' promoter region that creates an additional Sp1-type (CCACC box) promoter site. The SRD5A2 valine to leucine (V89L) polymorphism has been correlated with lower dihydroxytestosterone levels. We tested genotypes in 108 prostate cases and 167 controls along with samples (n = 340) from several different ethnic groups. The CYP17 A2 allele (combined A1/A2 and A2/A2 genotypes) occurred at a higher frequency in Caucasian patients with prostate cancer (70%) than in Caucasian clinical control urology patients (57%), suggesting that the A2 allele may convey increased risk for prostate cancer [odds ratio (OR) = 1.7, 95% confidence interval (CI) = 1.0-3.0]. Blacks and Caucasians had a similar frequency of the A2 genotype (16 and 17%, respectively) while Taiwanese had the highest frequency (27%). The SRD5A2 leucine genotype was most frequent in Taiwanese (28%), intermediate in Caucasians (8.5%) and lowest in Blacks (2.5%). Genotypes having a SRD5A2 leucine allele were somewhat more common in prostate cancer cases (56%) than in controls (49%) (OR = 1.4, 95% CI = 0.8-2.2) but this difference was not significant. These results support the hypothesis that some allelic variants of genes involved in androgen biosynthesis and metabolism may be associated with prostate cancer risk. PMID- 10469618 TI - Codon 72 polymorphism of p53 as a risk factor for patients with human papillomavirus-associated squamous intraepithelial lesions and invasive cancer of the uterine cervix. AB - Squamous intraepithelial lesions (SIL) and invasive cancer of the uterine cervix are thought to be a series of lesions derived from normal cervical squamous tissue. Infection by high risk human papillomavirus (HPV) and integration of viral DNA may initially lead normal cervical cells to become pre-malignant cells in SIL and result in cervical malignancies later on. High risk HPVs, including types 16 and 18, produce a viral protein, E6, which is required for viral replication in host cells. The E6 protein is able to bind to host p53 causing inactivation of its function through the mechanism of ubiquitin-dependent degradation. It has recently been reported that the extent of p53 dysfunction caused by HPVs depends on the status of a polymorphism at codon 72 of p53, Pro or Arg. In that study, it was demonstrated that a patient homozygous for the Arg allele had about a seven times higher risk of developing cervical cancer than a patient homozygous for Pro. In an attempt to confirm this result and elucidate whether this allelic deviation of the Arg genotype seen in invasive cervical cancer occurs in the pre-malignant lesion SIL, we analyzed 219 SIL and 101 invasive cancer samples from Japanese patients using a PCR-based assay. Samples from 88 SIL and 76 invasive cancers were identified as HPV-infected samples and used for further analyses. In these, the frequencies of Arg homozygotes were 31.8, 33.0 and 36.8% in controls, SIL and invasive cancer, respectively. The distributions of the different alleles of codon 72 (Pro/Pro, Pro/Arg and Arg/Arg) did not show significant differences between either control and SIL groups or control and invasive cancer groups. Also, no difference in the frequency of Arg/Arg genotype was detected even between the control and HSIL groups or control and invasive cancer infected with high risk HPVs groups. In conclusion, there was no obvious relationship between the Arg genotype at codon 72 of p53 and predisposition to HPV-associated cervical neoplasia. PMID- 10469619 TI - Anti-tumor-promoting activity of a polyphenolic fraction isolated from grape seeds in the mouse skin two-stage initiation-promotion protocol and identification of procyanidin B5-3'-gallate as the most effective antioxidant constituent. AB - Procyanidins present in grape seeds are known to exert anti-inflammatory, anti arthritic and anti-allergic activities, prevent skin aging, scavenge oxygen free radicals and inhibit UV radiation-induced peroxidation activity. Since most of these events are associated with the tumor promotion stage of carcinogenesis, these studies suggest that grape seed polyphenols and the procyanidins present therein could be anticarcinogenic and/or anti-tumor-promoting agents. Therefore, we assessed the anti-tumor-promoting effect of a polyphenolic fraction isolated from grape seeds (GSP) employing the 7,12-dimethylbenz[a]anthracene (DMBA) initiated and 12-O-tetradecanoylphorbol 13-acetate (TPA)-promoted SENCAR mouse skin two-stage carcinogenesis protocol as a model system. Following tumor initiation with DMBA, topical application of GSP at doses of 0.5 and 1.5 mg/mouse/application to the dorsal initiated mouse skin resulted in a highly significant inhibition of TPA tumor promotion. The observed anti-tumor-promoting effects of GSP were dose dependent and were evident in terms of a reduction in tumor incidence (35 and 60% inhibition), tumor multiplicity (61 and 83% inhibition) and tumor volume (67 and 87% inhibition) at both 0.5 and 1.5 mg GSP, respectively. Based on these results, we directed our efforts to separate and identify the individual polyphenols present in GSP and assess their antioxidant activity in terms of inhibition of epidermal lipid peroxidation. Employing HPLC followed by comparison with authentic standards for retention times in HPLC profiles, physiochemical properties and spectral analysis, nine individual polyphenols were identified as catechin, epicatechin, procyanidins B1-B5 and C1 and procyanidin B5-3'-gallate. Five of these individual polyphenols with evident structural differences, namely catechin, procyanidin B2, procyanidin B5, procyanidin C1 and procyanidin B5-3'-gallate, were assessed for antioxidant activity. All of them significantly inhibited epidermal lipid peroxidation, albeit to different levels. A structure-activity relationship study showed that with an increase in the degree of polymerization in polyphenol structure, the inhibitory potential towards lipid peroxidation increased. In addition, the position of linkage between inter-flavan units also influences lipid peroxidation activity; procyanidin isomers with a 4-6 linkage showed stronger inhibitory activity than isomers with a 4-8 linkage. A sharp increase in the inhibition of epidermal lipid peroxidation was also evident when a gallate group was linked at the 3'-hydroxy position of a procyanidin dimer. Procyanidin B5-3'-gallate showed the most potent antioxidant activity with an IC(50) of 20 microM in an epidermal lipid peroxidation assay. Taken together, for the first time these results show that grape seed polyphenols possess high anti-tumor-promoting activity due to the strong antioxidant effect of procyanidins present therein. In summary, grape seed polyphenols in general, and procyanidin B5-3'-gallate in particular, should be studied in more detail to be developed as cancer chemopreventive and/or anticarcinogenic agents. PMID- 10469620 TI - Induction of melanoma in TPras transgenic mice. AB - In order to study the oncogenesis of melanocytes, transgenic mouse lines were established that express a mutated human Ha-ras (TPras) gene in pigment producing cells. The ras transgenic mice exhibit an altered phenotype, including melanocytic hyperplasia and a muted agouti coat, indicative of hyperproliferative melanocytes. These mice and their wild-type littermates have been subjected to a variety of carcinogenesis protocols, including 7, 12-dimethylbenz-[a]anthracene (DMBA), 12-O-tetradecanoylphorbol-13-acetate (TPA) and UV radiation exposure. Topical DMBA treatment of TPras mice resulted in a high incidence of melanomas. Metastatic lesions were observed in skin, lungs and lymph nodes. TPA treatment of TPras mice induced a small number of papillomas but no nevi or melanomas. UV light exposures induced papillomas in negative littermate and melanomas in some albino TPras mice. These results show that melanocytes expressing an activated Ha ras in the TPras transgenic mice are susceptible to induction of melanoma by DMBA. PMID- 10469621 TI - Thapsigargin has similar effect on p53 protein response to benzo[a]pyrene-DNA adducts as TPA in mouse skin. AB - The level of p53 tumor suppressor protein increases in response to DNA damage caused by benzo[a]pyrene (B[a]P). The most used tumor promoter in the two step mouse skin carcinogenesis model, 12-O-tetradecanoylphorbol-13-acetate (TPA) decreases this response in mouse skin. In this study the effect of another promoter, thapsigargin was tested on B[a]P-induced p53 response using immunohistochemistry, western blotting and immunoelectron microscopy. We also studied the localization of p53 protein after treatments with BP and TPA or thapsigargin. Thapsigargin had a TPA-like effect on the acute induction of p53 protein related to benzo[a]pyrene-7, 8-diol-9,10-epoxide-DNA adducts in the skin of C57BL/6 mouse. After B[a]P treatment, there was slightly more putatively wild type p53 protein in nuclei than in cytoplasm of the cells. Neither TPA nor thapsigargin affected the localization of p53 protein. Since both compounds increase the level of intracellular calcium, the inhibition of the p53 response may depend on the level of intracellular calcium. Inhibition of the putatively genome-protecting increase in p53 protein may be one of the critical effects of tumor promoters. PMID- 10469622 TI - Induction of thioredoxin, thioredoxin reductase and glutaredoxin activity in mouse skin by TPA, a calcium ionophore and other tumor promoters. AB - We have measured the levels of thioredoxin, thioredoxin reductase and glutaredoxin enzyme activity in mouse skin following topical application of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C (PKC) activator and tumor promoter. The specific activity of thioredoxin and thioredoxin reductase in extracts from normal epidermis increased by 40 and 50%, respectively, after single or multiple application of TPA. Multiple applications (twice per week for 2 weeks) of TPA increased glutaredoxin activity by >300%. Induction of the proteins lasted several days. Other PKC activators, like 12-O retinoylphorbol 13-acetate, mezerein, 1-oleoyl-2-acetylglycerol and the calcium ionophore A23187, also induced all the enzyme activities. Phorbol and 4-O-methyl 12-O-tetradecanoylphorbol-13-acetate, weak activators of PKC, selectively induced the thioredoxin system only and did not influence glutaredoxin activity. Multiple applications of TPA to tumor initiated (7,12-dimethyl[a]benzanthracene-treated) skin resulted in elevated levels of both the thioredoxin and glutaredoxin systems when examined 6 days after the last phorbol ester treatment. Induction of thioredoxin, thioredoxin reductase and glutaredoxin activities by TPA and calcium ionophores may play a general role in the epigenetic mechanism of tumor promotion via thiol redox control mechanisms. PMID- 10469623 TI - Cigarette smoke induces direct DNA damage in the human B-lymphoid cell line Raji. AB - Human lymphoid cells (Raji) were exposed to water-soluble compounds from cigarette smoke (CS) generated in a smoking machine. DNA damage, as detected by alkaline single-cell microelectrophoresis (COMET assay), was induced in a time- and concentration-dependent manner in the cells. Most of the rapidly induced DNA damage was attributable to direct-acting compounds since cytochrome P450-related metabolic activities (ethoxy- and pentoxyresorufin-O-deethylases and coumarin-7 hydroxylase) were absent or very low. In addition, induction of DNA damage could be inhibited only slightly by beta-naphthoflavone and coumarin. Vitamin C enhanced DNA damage in Raji cells probably by redox cycling of catechol and hydroquinone present in CS implicating reactive oxygen intermediates as another source of DNA damage. N-acetylcysteine, a radical scavenger and glutathione precursor, reduced DNA damage in Raji cells when exposure to CS was followed by 2 h post-incubation in culture medium. Unrepaired DNA damage caused by CS persisted longer than gamma-irradiation-induced DNA damage. Among the CS constituents, acrolein, but not formaldehyde and acetaldehyde, induced DNA damage although less intensely than CS itself. At 50 and 100 microM concentrations, acrolein also inhibited repair of gamma- irradiation-induced DNA damage in the COMET assay. Inhibition of DNA synthesis by acrolein at 50 microM was demonstrated by an immunochemical assay for bromo-deoxyuridine incorporation; however, inhibition of a representative repair enzyme, 8-oxoguanosine hydrolase, by either CS or acrolein was not observed. The present results further confirm the presence of direct-acting genotoxic components and inhibitors of DNA repair in the gas phase of tobacco smoke, that may contribute to DNA damage and smoking-associated cancers of the upper aerodigestive tract. PMID- 10469624 TI - Benzylic hydroxylation of 1-methylpyrene and 1-ethylpyrene by human and rat cytochromes P450 individually expressed in V79 Chinese hamster cells. AB - Alkyl-substituted polycyclic aromatic hydrocarbons may be metabolized to highly reactive benzylic sulfuric acid esters via benzylic hydroxylation and subsequent sulfonation. We have studied the benzylic hydroxylation of 1-methylpyrene (MP), a hepatocarcinogen in rodents, and 1-ethylpyrene (EP), whose benzylic hydroxylation would produce a secondary alcohol (alpha-HEP), in contrast to the primary alcohol (alpha-HMP) formed from MP. The hydrocarbons were incubated with hepatic microsomal preparations from humans and rats, as well as with V79-derived cell lines engineered for the expression of individual cytochrome P450 (CYP) forms from human (1A1, 1A2, 1B1, 2A6, 2E1, 3A4) and rat (1A1, 1A2, 2B1). All microsomal systems and CYP-expressing cell lines used, but not CYP-deficient V79 cells, showed biotransformation of both hydrocarbons. Formation of the benzylic alcohol was detected in each case. alpha-HMP and its oxidation product, 1 pyrenylcarboxylic acid (COOH-P), accounted for a major part of the total amount of the metabolites formed from MP in the presence of human liver microsomes (38 64%) and cells expressing human 3A4, 2E1 or 1B1 (80-85%). Likewise, cells expressing human 1A1 showed a higher contribution of alpha-HMP and COOH-P to the total metabolites (45%) than cells expressing the orthologous enzyme of the rat (3%). EP was metabolized at a higher rate and with modified regioselectivity compared with MP, although omega-hydroxylation of the side chain was not detected with the cell lines and only accounted for a small percent of the biotransformation by the microsomal preparations. The highest contributions of alpha-HEP to the total metabolites from EP were detected with the cells expressing human 1A1, 1B1 and 3A4 (38-51%). alpha-HEP accounted for 16% of the metabolites formed in the presence of human hepatic microsomes. Thus, benzylic hydroxylation is a major initial step in the metabolism of MP and EP. This pathway appears to be even more important in humans than in rats. Previously, we had shown that the second step of the activation, the sulfonation of alpha-HMP and alpha-HEP, is also efficiently catalysed by various forms of human sulfotransferases. PMID- 10469625 TI - Molecular dosimetry of endogenous and ethylene oxide-induced N7-(2-hydroxyethyl) guanine formation in tissues of rodents. AB - The formation of N7-(2-hydroxyethyl)guanine (7-HEG) in DNA was investigated previously in target and non-target tissues of F-344 rats and B6C3F1 mice exposed to >/=ISOdia>/=10 p.p.m. concentrations of ethylene oxide (EO) using fluorescence linked high-performance liquid chromatography [V.E. Walker et al. (1992) Cancer Res., 52, 4238-4334]. In order to study the dose-responses for 7-HEG at lower exposures, a highly sensitive and specific gas chromatography coupled with high resolution mass spectrometry (GC-HRMS) assay was developed. DNA was extracted from liver, brain, lung and spleen of B6C3F1 mice and F-344 rats exposed to 0, 3, 10, 33 or 100 p.p.m. EO for 4 weeks (6 h/day, 5 days/week). Analysis of DNA from control rodents showed that endogenous 7-HEG varied from 0.2 +/- 0.1 to 0.3 +/- 0.2 pmol/micromol guanine in tissues of rats and mice. 7-HEG exhibited tissue- and species-specific dose-response relationships in EO-exposed animals. Linear dose-response relationships were evident in mouse liver, brain and spleen at exposures between 3 and 100 p.p.m. Mouse lung exhibited a slightly sublinear response between 33 and 100 p.p.m. EO. The relationships were linear in liver and spleen of rats between 3 and 100 p.p.m. EO, but were slightly sublinear in brain and lung between 33 and 100 p.p.m. EO. The number of 7-HEG adducts present in rats exposed to 3 p.p.m. EO was 5.3-12.5 times higher than endogenous 7-HEG in unexposed controls. In contrast, mice exposed to 3 p.p.m. EO only had 1.3- to 2.5 fold greater numbers of 7-HEG adducts. The factors driving the exposure-response relationships are also likely to affect carcinogenic and mutagenic responses of rodents to EO. Likewise, a better understanding of the relationships between 7 HEG derived from low exposures to EO and endogenously formed 7-HEG may be important for the accurate extrapolation of risk to humans. PMID- 10469626 TI - The promotion effect of anorectic drugs on aflatoxin B(1)-induced hepatic preneoplastic foci. AB - The ability of three extensively used anorectic drugs, namely fenfluramine (FN), fluoxetine (FX) and amphetamine (AM), to alter the development of aflatoxin B(1) (AFB(1))-induced gamma-glutamyl-positive (GGT(+)) preneoplastic liver foci was investigated in 135 male weanling F344 rats. Following AFB(1) administration, 15 rats were killed, while the rest were divided into four groups and fed diets containing either FN, FX, AM or control diet, with half of the animals in each group subsequently being killed at 4 weeks and half at 10 weeks. All three anorectic drugs as expected suppressed initial food intake, growth rate, body weight gain and food efficiency. They also tended to suppress body fat mass and to decrease plasma levels of T(3) and T(4). FN significantly (P < 0.05) increased GGT(+) foci number/cm(2) and number/cm(3), while FX significantly increased GGT(+) foci number/cm(2) and the volume fraction of foci. Histopathological staining also revealed that FN- and FX-treated animals had more serious morphological alterations in their liver tissue. In contrast, foci development was, if anything, suppressed by AM feeding. These results indicate that serotoninergic drugs (FN and FX), as opposed to dopaminergic drugs (AM), may have tumor promoter activity, at least for liver tissue. PMID- 10469627 TI - An esophagogastroduodenal anastomosis model for esophageal adenocarcinogenesis in rats and enhancement by iron overload. AB - The aim of this study is to establish a good animal model for esophageal adenocarcinoma (EAC) and to test the hypothesis that iron over-nutrition enhances EAC formation. With rats, esophagogastroduodenal anastomosis (EGDA) was accomplished by anastomosing the duodenum to the gastroesophageal junction. Iron supplementation was given by i.p. injection of iron dextran (4 mg Fe/kg/week). This model mimics the development of human EAC by introducing mixed reflux of gastric and duodenal contents. At 40 weeks after surgery, the body weight, food intake, hemoglobin, total serum iron, transferrin saturation, serum albumin, and plasma levels of alpha-tocopherol, gamma-tocopherol and retinol of the EGDA rats were not significantly different from those of the non-operated controls. The animals generally had only mild esophagitis, except that the area surrounding the anastomosis opening had more severe esophagitis. Columnar-lined esophagus (CLE), CLE with dysplasia, and EAC were diagnosed in 53.5, 34.9 and 25.6%, respectively, of the 43 rats. Intraperitoneal iron supplementation significantly enhanced esophageal lesions with CLE, CLE with dysplasia, and EAC to 78.0, 53. 7 and 53.7%, respectively, of the 41 rats. All the tumors were well-differentiated mucinous adenocarcinomas at the squamocolumnar junction area, where most iron deposition was observed. EGDA avoids nutritional problems seen in other animal models for EAC. We believe that direct anastomosis of squamous epithelium to columnar epithelium and mixed reflux of gastric and duodenal contents lead to the formation of CLE and EAC. With this model, we demonstrated that iron supplementation significantly enhanced EAC formation, suggesting that iron over nutrition could also be a risk factor for human EAC. PMID- 10469628 TI - Biotransformation of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK) in freshly isolated human lung cells. AB - Metabolism of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK) was characterized in human lung cells isolated from peripheral lung specimens obtained from 12 subjects during clinically indicated lobectomy. NNK biotransformation was assessed in preparations of isolated unseparated cells (cell digest), as well as in preparations enriched in alveolar type II cells, and alveolar macrophages. Metabolite formation was expressed as a percentage of the total recovered radioactivity from [5-(3)H]NNK and its metabolites per 10(6) cells per 24 h. 4-(Methylnitrosamino)-1-(3-pyridyl)-1 butanol (NNAL) was the major metabolite formed in all lung cell preparations examined, and its formation ranged from 0.50 to 13%/10(6) cells/24 h. Formation of alpha-carbon hydroxylation end-point metabolites (bioactivation) and pyridine N-oxidation metabolites (detoxification), ranged from non-detectable to 0.60% and from non-detectable to 1.5%/10(6) cells/24 h, respectively, reflecting a large degree of intercellular and inter-individual variability in NNK metabolism. Formation of the alpha-hydroxylation end-point metabolite 4-hydroxy-1-(3-pyridyl) 1-butanol (diol) was consistently higher in alveolar type II cells than in cell digest or alveolar macrophages (0.0146 +/- 0.0152, 0.0027 +/- 0.0037 and 0.0047 +/- 0.0063%/10(6) cells/24 h, respectively; n = 12; P < 0.05). SKF-525A was used to examine cytochrome P450 contributions to the biotransformation of NNK. SKF 525A inhibited keto reduction of NNK to NNAL by 85, 86 and 74% in cell digest, type II cells, and macrophages, respectively (means of 11 subjects, P < 0.05). Type II cell incubates treated with SKF-525A formed significantly lower amounts of total alpha-hydroxylation metabolites compared with type II cells without SKF 525A (0.0776 +/- 0.0841 versus 0.1694 +/- 0. 2148%/10(6) cells/24 h, respectively; n = 11; P < 0.05). The results of this first study examining NNK biotransformation in freshly isolated human lung cells indicate that NNK metabolism is subject to a large degree of inter-individual and intercellular variability, and suggest a role for P450s in human lung cell NNK metabolism. Both alveolar type II cells and alveolar macrophages may be potential target cells for NNK toxicity based on their alpha-carbon hydroxylation capabilities. In addition, carbonyl reduction of NNK to NNAL is SKF-525A sensitive in human lung cells. PMID- 10469629 TI - Nickel-induced transformation shifts the balance between HIF-1 and p53 transcription factors. AB - Nickel (Ni) compounds are potent carcinogens and can induce malignant transformation of rodent and human cells. In an attempt to unravel the molecular mechanisms of Ni-induced transformation we investigated transcriptional activity of hypoxia-inducible factor (HIF-1) and p53 tumor suppressor protein in Ni transformed cells. We demonstrated that the activity of HIF-1-responsive promoters was increased in Ni-transformed rodent cells resulting in the increased ratio between HIF-1- and p53-stimulated transcription. To further elucidate the roles of HIF-1 and p53 in Ni-induced transformation we used human osteosarcoma (HOS) cells and a Ni-transformed derivative, SA-8 cells. Since non-functional p53 was expressed in both HOS and SA-8 cells, acute Ni treatment induced HIF-1alpha protein and HIF-1-dependent transcription without affecting p53. In MCF-7 and A549, human cancer cells with the wild-type p53, both functional p53 and HIF 1alpha proteins accumulated following exposure to Ni. The induction of HIF-1alpha and wild-type p53 by Ni was detected after 6 h and was most pronounced by 24 h. These results suggest that acute Ni treatment causes accumulation of HIF-1alpha protein and simultaneous accumulation of wild-type, but not mutant, p53. We suggest that the induction of hypoxia-like conditions in Ni-treated cells with subsequent selection for increased HIF-1-dependent transcription is involved in Ni-induced carcinogenesis. PMID- 10469630 TI - CYP1A2 is not the primary enzyme responsible for 4-aminobiphenyl-induced hepatocarcinogenesis in mice. AB - 4-Aminobiphenyl (4-ABP), a potent carcinogen in rodents (liver cancer) and human (bladder cancer), is found as an environmental contaminant and in tobacco smoke. Hemoglobin adducts and lung DNA adducts of 4-ABP are found in tobacco smokers. In vitro metabolism studies with human and rat liver microsomes have shown that CYP1A2 is primarily responsible for catalyzing N-hydroxylation, the initial step in the metabolic activation of 4-ABP. To determine whether this P450 is a rate limiting pathway for hepatocarcinogenesis, CYP1A2-null mice were analyzed at 16 months of age and were compared with wild-type mice in their response to 4-ABP using the neonatal mouse bioassay and two different doses of the carcinogen. Overall differences in incidences of hepatocellular adenoma, carcinoma and preneoplastic foci were not significant between either genotypes or 4-ABP doses used, whereas small, but significant, differences were found for specific types of foci. These results suggest that while CYP1A2 levels may not be rate limiting for 4-ABP metabolism to produce tumors and foci, it may modulate the induction process of some types of liver foci in either a positive or negative manner. In vitro studies using CYP1A2-null and wild-type mouse liver microsomes revealed that CYP1A2 is not the sole P450 required for 4-ABP N-hydroxylation and that another, yet to be identified, P450 is likely to be involved. PMID- 10469631 TI - Exposure to flaxseed or its lignan component during different developmental stages influences rat mammary gland structures. AB - Reduction of the highly proliferative terminal end bud (TEB) structures in the developing mammary gland by differentiation to alveolar buds (ABs) and lobules has been suggested to be protective against mammary cancer. Flaxseed is high in alpha-linolenic acid (ALA) and secoisolariciresinol diglycoside (SDG). SDG is the precursor of mammalian lignans, which can affect mammary gland structures. Thus, the objective of this study was to determine the effect of lifetime, gestation and lactation or after-weaning exposure to 5 or 10% flaxseed or SDG and flaxseed oil components on the mammary gland structures of virgin female rat offspring at post-natal day 50. Lifetime or gestation and lactation exposure to flaxseed altered mammary gland structure development, whereas exposure to flaxseed after weaning had no effect. Lifetime or gestation and lactation exposure to 5% flaxseed caused endocrine changes, as suggested by delayed puberty onset and reduced number of estrous cycles. These changes reduced exposure to endogenous estrogens, leading to atrophy of mammary TEB structures. SDG, but not flaxseed oil, at the level found in 5% flaxseed produced similar effects as 5% flaxseed. This suggested that the lignans were the component in flaxseed responsible for the observed effects. Lifetime or gestation and lactation exposure to 10% flaxseed also caused endocrine changes, as suggested by early puberty onset and lengthened cycles due to prolonged estrus. This increased exposure to endogenous estrogens and stimulated mammary gland differentiation, as indicated by fewer TEBs and more ABs. Thus, lifetime or gestation and lactation exposure to 5 or 10% flaxseed induced structural changes in the mammary gland that may potentially reduce mammary cancer risk. PMID- 10469632 TI - A multihit, multistage model of chemical carcinogenesis. AB - Carcinogenesis involves the accumulation of genetic changes within a single cell. Tumor promotion functions in the initial clonal expansion of an initiated cell but is generally not considered to influence later stages. To investigate whether tumor promotion can influence later stages of carcinogenesis we developed a two hit 7, 12-dimethylbenz[a]anthracene (D) protocol designed to enrich for keratinocytes that contain at least two D-induced genetic alterations. FVB/N mice were initiated with D and promoted with 12-O-tetradecanoylphorbol-13-acetate (T) or treated with acetone (A) vehicle for 6 weeks. At 7 weeks after the start of promotion, but before visible papilloma development, groups of mice were treated with a second dose of D or A and 1 week later T promotion was resumed. D/T/A/T mice developed 2.8 papillomas/mouse and D/A/D/T mice demonstrated an additive tumor response and developed 5.8 papillomas/mouse. Importantly, D/T/D/T mice developed 12.4 papillomas/mouse, thereby demonstrating a synergistic tumor response compared with D/A/D/T and D/T/A/T mice. D/T/D/T papillomas exhibited increases in suprabasal S phase cells and keratin 13 expression when compared with D/T/A/T papillomas. D/T/D/T mice developed squamous cell carcinomas (SCCs) 10 weeks earlier than D/T/A/T mice and demonstrated a 96% malignancy incidence and 1.71 SCC/mouse compared with D/T/A/T mice, which demonstrated a 28% malignancy incidence and 0.32 SCC/mouse. Greater than 90% of D/T/A/T and D/T/D/T papillomas and SCCs contained mutant Ha-ras, while a normal Ha-ras allele persisted in all cases, indicating that a gene other than the remaining normal allele of Ha-ras was a target gene for the second D hit. These data demonstrate that: (i) promotion between the first and second hits has a profound outcome on carcinogenesis, presumably by increasing the probability that a second hit will occur in a previously initiated cell; (ii) continued promotion after the second hit is required for full expression of malignancy; (iii) the classic initiation promotion protocol can be extended to a multihit, multistage model. PMID- 10469633 TI - A deficiency in a 230 kDa DNA repair protein in fanconi anemia complementation group A cells is corrected by the FANCA cDNA. AB - Cells from individuals with the cancer-prone, inherited disorder Fanconi anemia (FA) are hypersensitive to DNA interstrand cross-linking agents and this hypersensitivity correlates with a defect in ability to repair this type of damage to their DNA. We have isolated a DNA endonuclease complex from the nuclei of normal human cells which is involved in repair of DNA interstrand cross-links and have shown that in FA complementation group A (FA-A) cells there is a defect in ability of this complex to incise DNA containing interstrand cross-links. In order to identify the specific protein(s) in this complex which is defective in FA-A cells, monoclonal antibodies (mAbs) were developed against proteins in the normal complex. One of these mAbs, which is against a protein with a molecular weight of approximately 230 kDa, completely inhibited the ability of the normal complex to incise cross-linked DNA. Western blot analysis has shown that there is a deficiency in this protein in FA-A cells. Electophoretic analysis has also indicated that there are reduced levels of this protein in FA-A compared with normal cells. Studies carried out utilizing FA-A cells which have been stably transduced with a retroviral vector expressing the FANCA cDNA have shown that the DNA repair defect in these cells has been corrected; levels of unscheduled DNA synthesis are at least as great as those of normal human cells. In addition, in the transduced cells the deficiency in the 230 kDa protein has been corrected, as determined by both western blot and electrophoretic analysis. These results indicate that the FANCA gene plays a role in the expression or stability of the 230 kDa protein. PMID- 10469634 TI - The cytotoxicity of DNA carboxymethylation and methylation by the model carboxymethylating agent azaserine in human cells. AB - Carboxymethylating agents are potential sources of endogenous DNA damage that have been proposed as possible contributors to gastrointestinal carcinogenesis. The cytotoxicity of the model DNA carboxymethylating agent azaserine was investigated in human cells. Expression of the DNA repair enzyme O(6) methylguanine-DNA methyltransferase (MGMT) did not affect sensitivity to the drug in two related Raji Burkitt's lymphoma cell lines. DNA mismatch repair-defective variants of Raji cells which display increased tolerance to DNA methylation damage were not selectively resistant to azaserine. Complementary results were obtained with a second carboxymethylating agent, potassium diazoacetate. In contrast, lymphoblastoid cell lines representative of each of the xeroderma pigmentosum complementation groups, including the variant, were all significantly more sensitive to azaserine than nucleotide excision repair-proficient cells. The hypersensitivity of XP cells was not due to systematic differences in the concentrations of intracellular thiol compounds or related thiol metabolizing enzymes. The data indicate that of the two types of potentially lethal DNA damage which azaserine introduces, carboxymethylated bases and O(6)-methylguanine, the former are repaired by nucleotide excision repair and are a more significant contributor to azaserine lethality in human cells. PMID- 10469635 TI - Overexpression of Ogg1 in mammalian cells: effects on induced and spontaneous oxidative DNA damage and mutagenesis. AB - Chinese hamster ovary cell lines (AA8 and AS52) were stably transfected to overexpress hOgg1 protein, the human DNA repair glycosylase for 7,8-dihydro-8 oxoguanine (8-oxoG). In the transfectants, the repair rate of 8-oxoG residues induced by either potassium bromate or the photosensitizer [R]-1-[(10-chloro-4 oxo-3-phenyl-4H-benzo[a]quinolizin-1-yl)-carbo nyl ]-2-pyrrolidinemethanolplus light was up to 3-fold more rapid than in the parental cells. However, the improved repair had little effect on the mutagenicity of potassium bromate in the guanine phosphoribosyl transferase (gpt) locus of the OGG1-transfected AS52 cells. The steady-state (background) levels of DNA base modifications sensitive to Fpg protein, which include 8-oxoG, in cells not exposed to a damaging agent were not reduced by the overexpression of Ogg1 protein. Moreover, the spontaneous mutation rates in the gpt locus were similar in OGG1-transformed and vector-only transformed cells. The results demonstrate the potential of Ogg1 protein to remove its substrate modifications from most of the chromosomal DNA. They indicate, on the other hand, that the Ogg1 protein alone may not be rate limiting for the repair of the residual substrate modifications observed in cells under normal growth conditions. PMID- 10469636 TI - Induction of cyclin-dependent kinase inhibitors and G(1) prolongation by the chemopreventive agent N-acetylcysteine. AB - Cyclin-dependent kinase (cdk) inhibitors, such as p16(INK4a) and p21(WAF1/CIP1), often inhibit G(1) cyclin kinases and result in G(1) arrest. It has been suggested that p21(WAF1/CIP1) may also play a role in other chemopreventive activities such as DNA repair, slowdown of DNA replication and induction of cellular differentiation. In this report we demonstrate that the antioxidant N acetylcysteine (NAC), a well-known chemopreventive agent, induces p16(INK4a) and p21(WAF1/CIP1) gene expression and prolongs cell-cycle transition through G(1) phase. A portion of the G(1) arrest by NAC is governed by p16(INK4a); it is independent of p53. NAC's usual mechanism of increasing intracellular glutathione level is not required for the G(1) arrest. An antioxidant whose action is limited to scavenging radicals, Trolox, does not induce G(1) arrest. Taken together, these results suggest a potential novel molecular basis for chemoprevention by NAC. PMID- 10469637 TI - Urinary bladder carcinogenicity of dimethylarsinic acid in male F344 rats. AB - The present study was conducted to determine the carcinogenicity of dimethylarsinic acid (DMA) administered to male F344 rats in a 2 year bioassay. A total of 144 rats (10 weeks old at the start) were divided into four groups of 36 rats each. Groups 1-4 received DMA (purity 100%) at concentrations of 200, 50, 12.5 and 0 p.p.m. in the drinking water, respectively, for 104 weeks. From weeks 97 to 104, urinary bladder tumors were observed in 12 of 31, eight of 31 and none of 33 in groups 1-3, respectively. No bladder tumors were observed in group 4. The present study demonstrated that long-term p. o. administration of DMA induced urinary bladder carcinomas in male F344 rats. Therefore, the results indicate that DMA is carcinogenic for the rat urinary bladder, which may be related to the human carcinogenicity of arsenicals. PMID- 10469638 TI - NAT2 slow acetylator genotype is associated with increased risk of lung cancer among non-smoking Chinese women in Singapore. AB - Among non-smokers, the factors resulting in lung carcinogenesis are poorly understood. We conducted a hospital-based case-control analysis of 294 Chinese women, of whom 217 were non-smokers, to evaluate the role of polymorphic N acetyltransferase (NAT2) as a susceptibility factor for the disease. The proportion of slow acetylator genotypes among non-smoking cases (n = 92) and controls (n = 125) was 38.0 and 24.0%, respectively [odds ratio (OR) 2.0, 95% confidence interval (CI) 1.1-3.7]. No effect of NAT2 genotype was seen among smokers. Among non-smokers, the effect was marked for adenocarcinomas (OR 2.1, 95% CI 1.1-4.0). As NAT2 activity is known to modify risk of arylamine-induced carcinogenesis, our results suggest that exposure to arylamines in the environment may play a role in risk of lung cancer among non-smokers. PMID- 10469639 TI - Aminoacyl-tRNA synthetases: a family of expanding functions. Mittelwihr, France, October 10-15, 1999. PMID- 10469640 TI - Crystal structure of a prokaryotic replication initiator protein bound to DNA at 2.6 A resolution. AB - The initiator protein (RepE) of F factor, a plasmid involved in sexual conjugation in Escherichia coli, has dual functions during the initiation of DNA replication which are determined by whether it exists as a dimer or as a monomer. A RepE monomer functions as a replication initiator, but a RepE dimer functions as an autogenous repressor. We have solved the crystal structure of the RepE monomer bound to an iteron DNA sequence of the replication origin of plasmid F. The RepE monomer consists of topologically similar N- and C-terminal domains related to each other by internal pseudo 2-fold symmetry, despite the lack of amino acid similarities between the domains. Both domains bind to the two major grooves of the iteron (19 bp) with different binding affinities. The C-terminal domain plays the leading role in this binding, while the N-terminal domain has an additional role in RepE dimerization. The structure also suggests that superhelical DNA induced at the origin of plasmid F by four RepEs and one HU dimer has an essential role in the initiation of DNA replication. PMID- 10469641 TI - Structure of the ligand-binding domain of oestrogen receptor beta in the presence of a partial agonist and a full antagonist. AB - Oestrogens exert their physiological effects through two receptor subtypes. Here we report the three-dimensional structure of the oestrogen receptor beta isoform (ERbeta) ligand-binding domain (LBD) in the presence of the phyto-oestrogen genistein and the antagonist raloxifene. The overall structure of ERbeta-LBD is very similar to that previously reported for ERalpha. Each ligand interacts with a unique set of residues within the hormone-binding cavity and induces a distinct orientation in the AF-2 helix (H12). The bulky side chain of raloxifene protrudes from the cavity and physically prevents the alignment of H12 over the bound ligand. In contrast, genistein is completely buried within the hydrophobic core of the protein and binds in a manner similar to that observed for ER's endogenous hormone, 17beta-oestradiol. However, in the ERbeta-genistein complex, H12 does not adopt the distinctive 'agonist' position but, instead, lies in a similar orientation to that induced by ER antagonists. Such a sub-optimal alignment of the transactivation helix is consistent with genistein's partial agonist character in ERbeta and demonstrates how ER's transcriptional response to certain bound ligands is attenuated. PMID- 10469642 TI - Phosphorylase recognition and phosphorolysis of its oligosaccharide substrate: answers to a long outstanding question. AB - Phosphorylases are key enzymes of carbohydrate metabolism. Structural studies have provided explanations for almost all features of control and substrate recognition of phosphorylase but one question remains unanswered. How does phosphorylase recognize and cleave an oligosaccharide substrate? To answer this question we turned to the Escherichia coli maltodextrin phosphorylase (MalP), a non-regulatory phosphorylase that shares similar kinetic and catalytic properties with the mammalian glycogen phosphorylase. The crystal structures of three MalP oligosaccharide complexes are reported: the binary complex of MalP with the natural substrate, maltopentaose (G5); the binary complex with the thio oligosaccharide, 4-S-alpha-D-glucopyranosyl-4-thiomaltotetraose (GSG4), both at 2.9 A resolution; and the 2.1 A resolution ternary complex of MalP with thio oligosaccharide and phosphate (GSG4-P). The results show a pentasaccharide bound across the catalytic site of MalP with sugars occupying sub-sites -1 to +4. Binding of GSG4 is identical to the natural pentasaccharide, indicating that the inactive thio compound is a close mimic of the natural substrate. The ternary MalP-GSG4-P complex shows the phosphate group poised to attack the glycosidic bond and promote phosphorolysis. In all three complexes the pentasaccharide exhibits an altered conformation across sub-sites -1 and +1, the site of catalysis, from the preferred conformation for alpha(1-4)-linked glucosyl polymers. PMID- 10469643 TI - Impaired retinal function and vitamin A availability in mice lacking retinol binding protein. AB - Retinol-binding protein (RBP) is the sole specific transport protein for retinol (vitamin A) in the circulation, and its single known function is to deliver retinol to tissues. Within tissues, retinol is activated to retinoic acid, which binds to nuclear receptors to regulate transcription of >300 diverse target genes. In the eye, retinol is also activated to 11-cis-retinal, the visual chromophore. We generated RBP knockout mice (RBP(-/-)) by gene targeting. These mice have several phenotypes. Although viable and fertile, they have reduced blood retinol levels and markedly impaired retinal function during the first months of life. The impairment is not due to developmental retinal defect. Given a vitamin A-sufficient diet, the RBP(-/-) mice acquire normal vision by 5 months of age even though blood retinol levels remain low. Deprived of dietary vitamin A, vision remains abnormal and blood retinol declines to undetectable levels. Another striking phenotype of the mutant mice is their abnormal retinol metabolism. The RBP(-/-) mice can acquire hepatic retinol stores, but these cannot be mobilized. Thus, their vitamin A status is extremely tenuous and dependent on a regular vitamin A intake. Unlike wild-type mice, serum retinol levels in adult RBP(-/-) animals become undetectable after only a week on a vitamin A-deficient diet and their retinal function rapidly deteriorates. Thus RBP is needed for normal vision in young animals and for retinol mobilization in times of insufficient dietary intake, but is otherwise dispensable for the delivery of retinol to tissues. PMID- 10469644 TI - Functional overlap between two classes of matrix-degrading proteases in wound healing. AB - Retarded wound healing was found in mice deficient in the serine protease precursor plasminogen, as well as in wild-type mice treated with the metalloprotease inhibitor galardin, but in both cases wound closure was ultimately completed in all mice within 60 days. The expression of several matrix metalloproteases in keratinocytes migrating to cover the wound was strongly enhanced by galardin treatment. However, when plasminogen-deficient mice were treated with galardin, healing was completely arrested and wound closure was not seen during an observation period of 100 days, demonstrating that protease activity is essential for skin wound healing. The requirement for both plasminogen deficiency and metalloprotease inhibition for complete inhibition of the healing process indicates that there is a functional overlap between the two classes of matrix-degrading proteases, probably in the dissection of the fibrin rich provisional matrix by migrating keratinocytes. Each class alone is capable of maintaining sufficient keratinocyte migration to regenerate the epidermal surface, although this function would normally be performed by both classes acting in parallel. Since there are strong similarities between the proteolytic mechanisms in wound healing and cancer invasion, these results predict that complete arrest of this latter process in therapeutic settings will require the use of inhibitors of both classes of proteases. PMID- 10469645 TI - Keratinocyte-specific ablation of Stat3 exhibits impaired skin remodeling, but does not affect skin morphogenesis. AB - To elucidate the biological role of Stat3 in the skin, conditional gene targeting using the Cre-loxP system was performed as germline Stat3 ablation leads to embryonic lethality. K5-Cre;Stat3(flox/-) transgenic mice, whose epidermal and follicular keratinocytes lack functional Stat3, were viable and the development of epidermis and hair follicles appeared normal. However, hair cycle and wound healing processes were severely compromised. Furthermore, mutant mice expressed sparse hair and developed spontaneously occurring ulcers with age. Growth factor dependent in vitro migration of Stat3-disrupted keratinocytes was impaired despite normal proliferative responses. We therefore conclude that Stat3 plays a crucial role in transducing a signal required for migration but not for proliferation of keratinocytes, and that Stat3 is essential for skin remodeling, including hair cycle and wound healing. PMID- 10469646 TI - The Drosophila STE20-like kinase misshapen is required downstream of the Frizzled receptor in planar polarity signaling. AB - The Drosophila misshapen (msn) gene is a member of the STE20 kinase family. We show that msn acts in the Frizzled (Fz) mediated epithelial planar polarity (EPP) signaling pathway in eyes and wings. Both msn loss- and gain-of-function result in defective ommatidial polarity and wing hair formation. Genetic and biochemical analyses indicate that msn acts downstream of fz and dishevelled (dsh) in the planar polarity pathway, and thus implicates an STE20-like kinase in Fz/Dsh mediated signaling. This demonstrates that seven-pass transmembrane receptors can signal via members of the STE20 kinase family in higher eukaryotes. We also show that Msn acts in EPP signaling through the JNK (Jun-N-terminal kinase) module as it does in dorsal closure. Although at the level of Fz/Dsh there is no apparent redundancy in this pathway, the downstream effector JNK/MAPK (mitogen-activated protein kinase) module is redundant in planar polarity generation. To address the nature of this redundancy, we provide evidence for an involvement of the related MAP kinases of the p38 subfamily in planar polarity signaling downstream of Msn. PMID- 10469647 TI - GIGANTEA: a circadian clock-controlled gene that regulates photoperiodic flowering in Arabidopsis and encodes a protein with several possible membrane spanning domains. AB - Flowering of Arabidopsis is promoted by long days and delayed by short days. Mutations in the GIGANTEA (GI) gene delay flowering under long days but have little or no effect under short days. We have now isolated the GI gene and show that it encodes a novel, putative membrane protein. By comparing the sequence of the Arabidopsis gene with that of a likely rice orthologue and by sequencing mutant alleles, we identify regions of the GI protein that are likely to be important for its function. We show that GI expression is regulated by the circadian clock with a peak in transcript levels 8-10 h after dawn. The timing, height and duration of this peak are influenced by daylength. We analysed the interactions between GI and the LHY, CCA1 and ELF3 genes, previously shown to affect daylength responses; we show that the rhythmic pattern of GI expression is altered in the elf3, CCA1-OX and lhy genotypes, and that CCA1 and LHY expression are reduced by gi mutations. Our results are consistent with the idea that GI plays an important role in regulating the expression of flowering time genes during the promotion of flowering by photoperiod. PMID- 10469648 TI - Early nuclear events in plant defence signalling: rapid gene activation by WRKY transcription factors. AB - Parsley WRKY proteins comprise a family of plant-specific zinc-finger-type factors implicated in the regulation of genes associated with pathogen defence. In vitro, these proteins bind specifically to functionally defined TGAC containing W box promoter elements within the Pathogenesis-Related Class10 (PR 10) genes. Here we present in vivo data demonstrating that WRKY1 is a transcriptional activator mediating fungal elicitor-induced gene expression by binding to W box elements. In situ RNA hybridization revealed that the WRKY1 gene is rapidly and locally activated in parsley leaf tissue around fungal infection sites. Transient expression studies in parsley protoplasts showed that a specific arrangement of W box elements in the WRKY1 promoter itself is necessary and sufficient for early activation and that WRKY1 binds to such elements. Our results demonstrate that WRKY transcription factors play an important role in the regulation of early defence-response genes including regulation of WRKY1. PMID- 10469649 TI - Mutations altering the moloney murine leukemia virus p12 Gag protein affect virion production and early events of the virus life cycle. AB - The p12 Gag protein of Moloney murine leukemia virus is a small polypeptide of unknown function, containing two proline-rich motifs. To determine its role in replication, we introduced a series of deletion and alanine-scanning substitution mutations throughout the p12 coding region of a proviral DNA, and characterized the phenotypes of the resulting mutant viruses. Complete deletion of p12 and mutations affecting the PPPY motif caused substantial reduction in the yield of virions and a modest reduction in Gag processing. Proteolytic cleavage of the R peptide from the cytoplasmic tail of the envelope protein TM was abolished in these mutants, suggesting that the PPPY motif is crucial for the viral protease to access the TM tail. The resulting virions were non-infectious, and unable to initiate DNA synthesis in infected cells. Mutants with alterations in both the N- and C-terminal portions of p12 exhibited a distinct phenotype. The production of virions and processing of Gag, Pol and Env precursors were normal. The viruses were able to direct synthesis of linear viral DNA, but there was almost no detectable circular DNAs or LTR-LTR junction. These data suggest that p12 plays a critical role in the early events of the virus life cycle. PMID- 10469650 TI - Molecular determinants of glycine receptor subunit assembly. AB - The inhibitory glycine receptor (GlyR) is a pentameric transmembrane protein composed of homologous alpha and beta subunits. Single expression of alpha subunits generates functional homo-oligomeric GlyRs, whereas the beta subunit requires a co-expressed alpha subunit to assemble into hetero-oligomeric channels of invariant stoichiometry (alpha(3)beta(2)). Here, we identified eight amino acid residues within the N-terminal region of the alpha1 subunit that are required for the formation of homo-oligomeric GlyR channels. We show that oligomerization and N-glycosylation of the alpha1 subunit are required for transit from the endoplasmic reticulum to the Golgi apparatus and later compartments, and that addition of simple carbohydrate side chains occurs prior to GlyR subunit assembly. Our data are consistent with both intersubunit surface and conformational differences determining the different assembly behaviour of GlyR alpha and beta subunits. PMID- 10469651 TI - PKA-mediated phosphorylation of the human K(ATP) channel: separate roles of Kir6.2 and SUR1 subunit phosphorylation. AB - ATP-sensitive potassium (K(ATP)) channels play important roles in many cellular functions such as hormone secretion and excitability of muscles and neurons. Classical ATP-sensitive potassium (K(ATP)) channels are heteromultimeric membrane proteins comprising the pore-forming Kir6.2 subunits and the sulfonylurea receptor subunits (SUR1 or SUR2). The molecular mechanism by which hormones and neurotransmitters modulate K(ATP) channels via protein kinase A (PKA) is poorly understood. We mutated the PKA consensus sequences of the human SUR1 and Kir6.2 subunits and tested their phosphorylation capacities in Xenopus oocyte homogenates and in intact cells. We identified the sites responsible for PKA phosphorylation in the C-terminus of Kir6.2 (S372) and SUR1 (S1571). Kir6.2 can be phosphorylated at its PKA phosphorylation site in intact cells after G-protein (Gs)-coupled receptor or direct PKA stimulation. While the phosphorylation of Kir6.2 increases channel activity, the phosphorylation of SUR1 contributes to the basal channel properties by decreasing burst duration, interburst interval and open probability, and also increasing the number of functional channels at the cell surface. Moreover, the effect of PKA could be mimicked by introducing negative charges in the PKA phosphorylation sites. These data demonstrate direct phosphorylation by PKA of the K(ATP) channel, and may explain the mechanism by which Gs-coupled receptors stimulate channel activity. Importantly, they also describe a model of heteromultimeric ion channels in which there are functionally distinct roles of the phosphorylation of the different subunits. PMID- 10469652 TI - Steady-state free Ca(2+) in the yeast endoplasmic reticulum reaches only 10 microM and is mainly controlled by the secretory pathway pump pmr1. AB - Over recent decades, diverse intracellular organelles have been recognized as key determinants of Ca(2+) signaling in eukaryotes. In yeast however, information on intra-organellar Ca(2+) concentrations is scarce, despite the demonstrated importance of Ca(2+) signals for this microorganism. Here, we directly monitored free Ca(2+) in the lumen of the endoplasmic reticulum (ER) of yeast cells, using a specifically targeted version of the Ca(2+)-sensitive photoprotein aequorin. Ca(2+) uptake into the yeast ER displayed characteristics distinctly different from the mammalian ER. At steady-state, the free Ca(2+) concentration in the ER lumen was limited to approximately 10 microM, and ER Ca(2+) sequestration was insensitive to thapsigargin, an inhibitor specific for mammalian ER Ca(2+) pumps. In pmr1 null mutants, free Ca(2+) in the ER was reduced by 50%. Our findings identify the secretory pathway pump Pmr1, predominantly localized in the Golgi, as a major component of ER Ca(2+) uptake activity in yeast. PMID- 10469653 TI - Pathological missense mutations of neural cell adhesion molecule L1 affect homophilic and heterophilic binding activities. AB - Mutations in the gene for neural cell adhesion molecule L1 (L1CAM) result in a debilitating X-linked congenital disorder of brain development. At the neuronal cell surface L1 may interact with a variety of different molecules including itself and two other CAMs of the immunoglobulin superfamily, axonin-1 and F11. However, whether all of these interactions are relevant to normal or abnormal development has not been determined. Over one-third of patient mutations are single amino acid changes distributed across 10 extracellular L1 domains. We have studied the effects of 12 missense mutations on binding to L1, axonin-1 and F11 and shown for the first time that whereas many mutations affect all three interactions, others affect homophilic or heterophilic binding alone. Patient pathology is therefore due to different types of L1 malfunction. The nature and functional consequence of mutation is also reflected in the severity of the resultant phenotype with structural mutations likely to affect more than one binding activity and result in early mortality. Moreover, the data indicate that several extracellular domains of L1 are required for homophilic and heterophilic interactions. PMID- 10469654 TI - STAT5 as a molecular regulator of proliferation, differentiation and apoptosis in hematopoietic cells. AB - Signal transducers and activators of transcription (STATs) play key roles in growth factor-mediated intracellular signal transduction. In the present study using a constitutively active STAT5 mutant, we show that STAT5 has pleiotropic functions regulating cell proliferation, differentiation and apoptosis in an IL-3 dependent Ba/F3 cell line. The mutant STAT5 possessed constitutive tyrosine phosphorylation and DNA binding activity, induced expression of bcl-xL and pim-1 in the absence of IL-3 in Ba/F3 cells, and rendered Ba/F3 cells factor independent. Unexpectedly, IL-3 treatment of the factor-independent Ba/F3 cells expressing the constitutively active STAT5 resulted in apoptosis within 24 h, or differentiation followed by cell death. In these cells, mRNA expression of growth inhibitory genes downstream of STAT5 such as CIS, JAB/SOCS-1/SSI-1, and p21(WAF1/Cip1) was highly induced, correlating with prolonged hyper phosphorylation of the mutant STAT5 after IL-3 stimulation. Of the STAT5 regulated genes, we found that constitutive expression of JAB/SOCS-1/SSI-1 was sufficient to induce apoptosis of Ba/F3 cells, while p21(WAF1/Cip1) could induce differentiation of these cells. In contrast, constitutive expression of pim-1 was sufficient to induce IL-3-independent growth of Ba/F3 cells. These findings suggest that a single transcription factor regulates cell fate by varying the intensity and duration of the expression of a set of target genes. PMID- 10469655 TI - NF-kappaB p105 is a target of IkappaB kinases and controls signal induction of Bcl-3-p50 complexes. AB - The NF-kappaB precursor p105 has dual functions: cytoplasmic retention of attached NF-kappaB proteins and generation of p50 by processing. It is poorly understood whether these activities of p105 are responsive to signalling processes that are known to activate NF-kappaB p50-p65. We propose a model that p105 is inducibly degraded, and that its degradation liberates sequestered NF kappaB subunits, including its processing product p50. p50 homodimers are specifically bound by the transcription activator Bcl-3. We show that TNFalpha, IL-1beta or phorbolester (PMA) trigger rapid formation of Bcl-3-p50 complexes with the same kinetics as activation of p50-p65 complexes. TNF-alpha-induced Bcl 3-p50 formation requires proteasome activity, but is independent of p50-p65 released from IkappaBalpha, indicating a pathway that involves p105 proteolysis. The IkappaB kinases IKKalpha and IKKbeta physically interact with p105 and inducibly phosphorylate three C-terminal serines. p105 is degraded upon TNF-alpha stimulation, but only when the IKK phospho-acceptor sites are intact. Furthermore, a p105 mutant, lacking the IKK phosphorylation sites, acts as a super-repressor of IKK-induced NF-kappaB transcriptional activity. Thus, the known NF-kappaB stimuli not only cause nuclear accumulation of p50-p65 heterodimers but also of Bcl-3-p50 and perhaps further transcription activator complexes which are formed upon IKK-mediated p105 degradation. PMID- 10469656 TI - The nucleosomal response associated with immediate-early gene induction is mediated via alternative MAP kinase cascades: MSK1 as a potential histone H3/HMG 14 kinase. AB - The nucleosomal response refers to the rapid phosphorylation of histone H3 on serine 10 and HMG-14 on serine 6 that occurs concomitantly with immediate-early (IE) gene induction in response to a wide variety of stimuli. Using antibodies against the phosphorylated residues, we show that H3 and HMG-14 phosphorylation is mediated via different MAP kinase (MAPK) cascades, depending on the stimulus. The nucleosomal response elicited by TPA is ERK-dependent, whereas that elicited by anisomycin is p38 MAPK-dependent. In intact cells, the nucleosomal response can be selectively inhibited using the protein kinase inhibitor H89. MAPK activation and phosphorylation of transcription factors are largely unaffected by H89, whereas induction of IE genes is inhibited and its characteristics markedly altered. MSK1 is considered the most likely kinase to mediate this response because (i) it is activated by both ERK and p38 MAPKs; (ii) it is an extremely efficient kinase for HMG-14 and H3, utilizing the physiologically relevant sites; and (iii) its activity towards H3/HMG-14 is uniquely sensitive to H89 inhibition. Thus, the nucleosomal response is an invariable consequence of ERK and p38 but not JNK/SAPK activation, and MSK1 potentially provides a link to complete the circuit between cell surface and nucleosome. PMID- 10469657 TI - The oxidative refolding of hen lysozyme and its catalysis by protein disulfide isomerase. AB - The oxidative refolding of hen lysozyme has been studied by a variety of time resolved biophysical methods in conjunction with analysis of folding intermediates using reverse-phase HPLC. In order to achieve this, refolding conditions were designed to reduce aggregation during the early stages of the folding reaction. A complex ensemble of relatively unstructured intermediates with on average two disulfide bonds is formed rapidly from the fully reduced protein after initiation of folding. Following structural collapse, the majority of molecules slowly form the four-disulfide-containing fully native protein via rearrangement of a highly native-like, kinetically trapped intermediate, des-[76 94], although a significant population (approximately 30%) appears to fold more quickly via other three-disulfide intermediates. The folding catalyst PDI increases dramatically both yields and rates of lysozyme refolding, largely by facilitating the conversion of des-[76-94] to the native state. This suggests that acceleration of the folding rate may be an important factor in avoiding aggregation in the intracellular environment. PMID- 10469658 TI - Control of glycosylation of MHC class II-associated invariant chain by translocon associated RAMP4. AB - Protein translocation across the membrane of the endoplasmic reticulum (ER) proceeds through a proteinaceous translocation machinery, the translocon. To identify components that may regulate translocation by interacting with nascent polypeptides in the translocon, we used site-specific photo-crosslinking. We found that a region C-terminal of the two N-glycosylation sites of the MHC class II-associated invariant chain (Ii) interacts specifically with the ribosome associated membrane protein 4 (RAMP4). RAMP4 is a small, tail-anchored protein of 66 amino acid residues that is homologous to the yeast YSY6 protein. YSY6 suppresses a secretion defect of a secY mutant in Escherichia coli. The interaction of RAMP4 with Ii occurred when nascent Ii chains reached a length of 170 amino acid residues and persisted until Ii chain completion, suggesting translocational pausing. Site-directed mutagenesis revealed that the region of Ii interacting with RAMP4 contains essential hydrophobic amino acid residues. Exchange of these residues for serines led to a reduced interaction with RAMP4 and inefficient N-glycosylation. We propose that RAMP4 controls modification of Ii and possibly also of other secretory and membrane proteins containing specific RAMP4-interacting sequences. Efficient or variable glycosylation of Ii may contribute to its capacity to modulate antigen presentation by MHC class II molecules. PMID- 10469659 TI - Different import pathways through the mitochondrial intermembrane space for inner membrane proteins. AB - Earlier work on the protein import system of yeast mitochondria has identified two soluble 70 kDa protein complexes in the intermembrane space. One complex contains the essential proteins Tim9p and Tim10p and mediates transport of cytosolically-made metabolite carrier proteins from the outer to the inner membrane. The other complex contains the non-essential proteins Tim8p and Tim13p as well as loosely associated Tim9p; its function was unclear, but it interacted structurally or functionally with the Tim9p-Tim10p complex. We now show that the two 70 kDa complexes each mediate the import of a different subset of integral inner membrane proteins and that they can transfer these proteins to one of three different membrane insertion sites: the TIM22 complex, the TIM23 complex or an as yet uncharacterized insertion site. Yeast mitochondria thus use multiple pathways for escorting hydrophobic inner membrane proteins across the aqueous intermembrane space. PMID- 10469660 TI - Mammalian TAF(II)30 is required for cell cycle progression and specific cellular differentiation programmes. AB - The two alleles of the 30 kDa TATA-binding protein associated factor (TAF(II)30) gene, have been targeted by homologous recombination in murine F9 embryonal carcinoma cells and subsequently disrupted using a Cre recombinase-loxP strategy. The TAF(II)30-null cells are not viable, but are rescued by the expression of human TAF(II)30. Cells lacking TAF(II)30 are blocked in G(1)/G(0) phase of the cell cycle and undergo apoptosis. In agreement with the G(1) arrest phenotype, the expression of cyclin E is impaired and the retinoblastoma protein is hypophosphorylated in the TAF(II)30-null cells. Interestingly, retinoic acid (RA) treatment prevented TAF(II)30-null cell death and induced primitive endodermal differentiation. In contrast, the RA- and cAMP-induced parietal endodermal differentiation was impaired in the TAF(II)30-null cells. Thus, TAF(II)30 is not indispensable for class II gene transcription in general, but seems to be required for the expression of a subset of genes. PMID- 10469661 TI - DNA binding site selection by RNA polymerase II TAFs: a TAF(II)250-TAF(II)150 complex recognizes the initiator. AB - Basal transcription factor TFIID comprises the TATA-box-binding protein, TBP, and associated factors, the TAF(II)s. Previous studies have implicated TAF(II)250 and TAF(II)150 in core promoter selectivity of RNA polymerase II. Here, we have used a random DNA binding site selection procedure to identify target sequences for these TAFs. Individually, neither TAF(II)250 nor TAF(II)150 singles out a clearly constrained DNA sequence. However, a TAF(II)250-TAF(II)150 complex selects sequences that match the Initiator (Inr) consensus. When in a trimeric complex with TBP, these TAFs select Inr sequences at the appropriate distance from the TATA-box. Point mutations that inhibit binding of the TAF(II)250-TAF(II)150 complex also impair Inr function in reconstituted basal transcription reactions, underscoring the functional relevance of Inr recognition by TAFs. Surprisingly, the precise DNA sequence at the start site of transcription influences transcriptional regulation by the upstream activator Sp1. Finally, we found that TAF(II)150 specifically binds to four-way junction DNA, suggesting that promoter binding by TFIID may involve recognition of DNA structure as well as primary sequence. Taken together, our results establish that TAF(II)250 and TAF(II)150 bind the Inr directly and that Inr recognition can determine the responsiveness of a promoter to an activator. PMID- 10469662 TI - Control of variant surface glycoprotein gene-expression sites in Trypanosoma brucei. AB - Trypanosoma brucei has 20 similar telomeric-expression sites for variant surface glycoprotein genes. Expression sites appear to be controlled at the level of transcription initiation, resulting in only one site being active at any time. Switching between expression sites occurs at a low rate. To analyse the switching mechanism, we used trypanosomes with two expression sites tagged with two different drug-resistance genes and selected these on agarose plates containing both drugs. Double-resistant clones arose at a low frequency of 10(-7) per cell, but these behaved as if they rapidly switched between the two tagged expression sites and lost double resistance in the absence of selection. Using in situ hybridization we found that only 10% of the double-resistant cells had two fluorescent spots corresponding to transcribed expression sites. Our results suggest that: (i) a double expressor is not a stable intermediate in expression site switching; (ii) expression sites are not independently switched on and off; and (iii) expression sites can be in a 'pre-active' silent state from which they can be readily activated. PMID- 10469663 TI - A conformational switch at the 3' end of a plant virus RNA regulates viral replication. AB - 3' untranslated regions of alfamo- and ilar-virus RNAs fold into a series of stem loop structures to which the coat protein binds with high affinity. This binding plays a role in initiation of infection ('genome activation') and has been thought to substitute for a tRNA-like structure that is found at the 3' termini of related plant viruses. We propose the existence of an alternative conformation of the 3' ends of alfamo- and ilar-virus RNAs, including a pseudoknot. Based on (i) phylogenetic comparisons, (ii) in vivo and in vitro functional analyses of mutants in which the pseudoknot has been disrupted or restored by compensatory mutations, (iii) competition experiments between coat protein and viral replicase, and (iv) investigation of the effect of magnesium, we demonstrate that this pseudoknot is required for replication of alfalfa mosaic virus. This conformation resembles the tRNA-like structure of the related bromo- and cucumo viruses. A low but specific interaction with yeast CCA-adding enzyme was found. The existence of two mutually exclusive conformations for the 3' termini of alfamo- and ilar-virus RNAs could enable the virus to switch from translation to replication and vice versa. The role of coat protein in this modulation and in genome activation is discussed. PMID- 10469664 TI - Translation driven by an eIF4G core domain in vivo. AB - Most eukaryotic mRNAs possess a 5' cap structure (m(7)GpppN) and a 3' poly(A) tail which promote translation initiation by binding the eukaryotic translation initiation factor (eIF)4E and the poly(A) binding protein (PABP), respectively. eIF4G can bridge between eIF4E and PABP, and-through eIF3-is thought to establish a link to the small ribosomal subunit. We fused the C-terminal region of human eIF4GI lacking both the eIF4E- and PABP-binding sites, to the IRE binding protein IRP-1. This chimeric protein suffices to direct the translation of the downstream cistron of bicistronic mRNAs bearing IREs in their intercistronic space in vivo. This function is preserved even when translation via the 5' end is inhibited. Deletion analysis defined the conserved central domain (amino acids 642-1091) of eIF4G as an autonomous 'ribosome recruitment core' and implicated eIF4A as a critical binding partner. Our data reveal the sufficiency of the conserved eIF4G ribosome recruitment core to drive productive mRNA translation in living cells. The C-terminal third of eIF4G is dispensable, and may serve as a regulatory domain. PMID- 10469665 TI - Natural synthesis of a DNA-binding protein from the C-terminal domain of DNA gyrase A in Borrelia burgdorferi. AB - We have identified a 34 kDa DNA-binding protein with an HU-like activity in the Lyme disease spirochete Borrelia burgdorferi. The 34 kDa protein is translated from an abundant transcript initiated within the gene encoding the A subunit of DNA gyrase. Translation of the 34 kDa protein starts at residue 499 of GyrA and proceeds in the same reading frame as full-length GyrA, resulting in an N terminal-truncated protein. The 34 kDa GyrA C-terminal domain, although not homologous, substitutes for HU in the formation of the Type 1 complex in Mu transposition, and complements an HU-deficient strain of Escherichia coli. This is the first example of constitutive expression of two gene products in the same open reading frame from a single gene in a prokaryotic cellular system. PMID- 10469666 TI - Escherichia coli SeqA protein affects DNA topology and inhibits open complex formation at oriC. AB - Chromosome replication in Escherichia coli is initiated by the DnaA protein. Binding of DnaA to the origin, oriC, followed by formation of an open complex are the first steps in the initiation process. Based on in vivo studies the SeqA protein has been suggested to function negatively in the initiation of replication, possibly by inhibiting open complex formation. In vitro studies have shown that SeqA inhibits oriC-dependent replication. Here we show by KMnO(4) probing that SeqA inhibits open complex formation. The inhibition was not caused by prevention of DnaA binding to the oriC plasmids, indicating that SeqA prevented strand separation in oriC either directly, by interacting with the AT rich region, or indirectly, by changing the topology of the oriC plasmids. SeqA was found to restrain the negative supercoils of the oriC plasmid. In comparison with the effect of HU on plasmid topology, SeqA seemed to act more cooperatively. It is likely that the inhibition of open complex formation is caused by the effect of SeqA on the topology of the plasmids. SeqA also restrained the negative supercoils of unmethylated oriC plasmids, which do not bind SeqA specifically, suggesting that the effect on topology is not dependent on binding of SeqA to a specific sequence in oriC. PMID- 10469667 TI - Strand opening by the UvrA(2)B complex allows dynamic recognition of DNA damage. AB - Repair proteins alter the local DNA structure during nucleotide excision repair (NER). However, the precise role of DNA melting remains unknown. A series of DNA substrates containing a unique site-specific BPDE-guanine adduct in a region of non-complementary bases were examined for incision by the Escherichia coli UvrBC endonuclease in the presence or absence of UvrA. UvrBC formed a pre-incision intermediate with a DNA substrate containing a 6-base bubble structure with 2 unpaired bases 5' and 3 unpaired bases 3' to the adduct. Formation of this bubble served as a dynamic recognition step in damage processing. UvrB or UvrBC may form one of three stable repair intermediates with DNA substrates, depending upon the state of the DNA surrounding the modified base. The dual incisions were strongly determined by the distance between the adduct and the double-stranded-single stranded DNA junction of the bubble, and required homologous double-stranded DNA at both incision sites. Remarkably, in the absence of UvrA, UvrBC nuclease can make both 3' and 5' incisions on substrates with bubbles of 3-6 nucleotides, and an uncoupled 5' incision on bubbles of >/=>/=10 nucleotides. These data support the hypothesis that the E.coli and human NER systems recognize and process DNA damage in a highly conserved manner. PMID- 10469668 TI - Advances in unstable angina: the role of low molecular weight heparins. Introduction and Chairmen's summary. PMID- 10469669 TI - Pathogenic mechanisms in unstable angina. PMID- 10469670 TI - Management of unstable angina based on considerations of aetiology. PMID- 10469671 TI - New antithrombotic and antiplatelet treatment. PMID- 10469672 TI - Low molecular weight heparin (enoxaparin) in the management of unstable angina: the ESSENCE study. Efficacy and Safety of Subcutaneous Enoxaparin in Non-Q wave Coronary Events. PMID- 10469673 TI - Low molecular weight heparins (enoxaparin) in the management of unstable angina: the TIMI studies. Thrombolysis in Myocardial Infarction. PMID- 10469674 TI - Optimising outcomes: socioeconomic perspective. PMID- 10469676 TI - Severe OHSS: An 'epidemic' of severe OHSS: a price we have to pay? PMID- 10469677 TI - Severe OHSS: patients should be allowed to weigh the morbidity of OHSS against the benefits of parenthood. PMID- 10469678 TI - Spermatid injection: reconsidering spermatid injection. PMID- 10469679 TI - What are the clinical benefits of recombinant gonadotrophins?: the development of recombinant FSH (Puregon((R))): a scientific business. PMID- 10469680 TI - Positive outcome after preimplantation diagnosis of aneuploidy in human embryos. AB - usromosomal abnormalities are responsible for a great deal of embryo wastage, which is reflected, at least partially, in decreased implantation and increased miscarriage in older women. To address this problem the transfer of only chromosomally normal embryos previously selected by preimplantation genetic diagnosis (PGD) has been proposed. We designed a multi-centre in-vitro fertilization (IVF) study to compare controls and a test group that underwent embryo biopsy and PGD for aneuploidy. Patients were matched retrospectively, but blindly, for average maternal age, number of previous IVF cycles, duration of stimulation, oestradiol concentrations on day +1, and average mature follicles. All these parameters were similar in test and control groups. Only embryos classified as normal for those chromosomes were transferred after PGD. The results showed that the rates of fetal heart beat (FHB)/embryo transferred between the control and test groups were similar. However, spontaneous abortions, measured as FHB aborted/FHB detected, decreased after PGD (P < 0.05), and ongoing pregnancies and delivered babies increased (P < 0.05) in the PGD group of patients. Two conclusions were obtained: (i) PGD of aneuploidy reduced embryo loss after implantation; (ii) implantation rates were not significantly improved, but the proportion of ongoing and delivered babies was increased. PMID- 10469681 TI - Gonadotrophin and testosterone suppression by 7alpha-methyl-19-nortestosterone acetate administered by subdermal implant to healthy men. AB - The synthetic androgen 7alpha-methyl-19-nortestosterone (MENT) is a potent suppressor of gonadotrophin that has several advantages for long term administration to normal or hypoandrogenic men. The aim of this study was to examine MENT serum concentrations following subdermal insertion of MENT acetate (MENT Ac) implants and their effects on gonadotrophins, testosterone, dihydrotestosterone (DHT), sex hormone-binding globulin, prostate specific antigen and insulin-like growth factor-1 serum concentrations in normal men. A total of 45 healthy men were recruited at three clinics. Each subject received one, two or four implants for 28 days. Serum samples were obtained before insertion and on days 8, 15, 22, 29, 36 and 43 after implant insertion. The average daily dose delivered in vivo by one implant was approximately 500 microg. One, two or four MENT Ac implants produced dose dependent and sustained serum MENT concentrations for the entire duration of treatment of 0.7 +/- 0.1, 1.2 +/- 0.1 and 2.0 +/- 0.1 nmol/l respectively. This treatment induced a dose dependent decrease in gonadotrophin and androgen serum levels. Two and four implants induced maximal suppression that was maintained throughout treatment and was completely reversed after removal of the implants. The mean decreases were 93 +/- 1% for testosterone, 80 +/- 3% for DHT, 97 +/- 1% for luteinizing hormone and 95 +/- 1% for follicle stimulating hormone. No serious adverse reactions were reported by the volunteers and no consistent changes in clinical chemistry and haematology were found. These results indicate that MENT Ac implants are an efficient way of MENT administration and confirm the potent gonadotrophin and androgen suppressive effect of this drug. PMID- 10469682 TI - Recombinant versus urinary follicle stimulating hormone for ovarian stimulation in assisted reproduction. AB - The recent availability of recombinant follicle stimulating hormone (rFSH), with its high level of purity and batch-to-batch consistency has made it an attractive alternative to urinary FSH (uFSH) for ovarian stimulation. Several trials have compared the two preparations, but none had sufficient power to detect a clinically meaningful difference in pregnancy rates. The purpose of this study was to determine the clinical pregnancy rates per started cycle by pooling data from randomized trials which compared the use of rFSH and uFSH in treatment cycles using in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). A thorough search of the literature identified 12 trials which met the inclusion criteria. In four trials, both IVF and ICSI were performed, in seven trials only IVF was performed and in one trial only ICSI was performed. Data were extracted and pooled using the principles of meta-analysis. There was no significant heterogeneity of treatment effect across the trials. The common odds ratio and the risk difference (and their 95% confidence intervals), obtained by pooling the data using a fixed effects model, were 1.20 (1.02-1.42) and 3.7% (0.5 6.9%) respectively, in favour of rFSH. The pregnancy rate with the alpha preparation of rFSH was statistically significantly higher than with uFSH in IVF cycles. The overall conclusion from this meta-analysis is that the use of rFSH in assisted reproduction is preferred over uFSH. PMID- 10469683 TI - Adipocyte insulin action following ovulation in polycystic ovarian syndrome. AB - The role of anovulation and insulin resistance in the pathogenesis of polycystic ovarian syndrome (PCOS) remains to be determined. The aim of this study was to investigate whether the metabolic abnormality of insulin resistance in PCOS reflects, rather than causes, the ovarian dysfunction. Eight subjects with classical PCOS were studied on two occasions. Adipocyte insulin sensitivity together with hormonal and metabolic changes were investigated in patients with PCOS following prolonged amenorrhoea and then again in the early follicular phase after ovulation. Insulin receptor binding in amenorrhoeic subjects with PCOS was low at 0.78 +/- 0.08% and this increased to 1.18 +/- 0.19% after an ovulatory cycle (P < 0.05). Maximal insulin stimulated 3-O-methylglucose uptake was 0.70 +/ 0. 14 during amenorrhoea and increased to 1.08 +/- 0.25 pmol/10 cm(2) cell membrane (P < 0.05). Plasma testosterone fell (4.0 +/- 0.4 to 2. 3 +/- 0.2 nmol/l; P < 0.001), luteinizing hormone fell (17.6 +/- 2.3 to 6.7 +/- 0.8 IU/l; P < 0.001) but plasma insulin concentrations remained unchanged following ovulation (14.6 +/- 1.9 and 15.7 +/- 3. 8 pmol/l during amenorrhoea and after ovulation respectively). The results of this study suggest that chronic anovulation per se appears to modify the factors contributing to cellular insulin resistance seen in PCOS. PMID- 10469684 TI - Endocrine features of polycystic ovary syndrome in a random population sample of 14-16 year old adolescents. AB - Hospital based studies have shown that oligomenorrhoeic adolescents have high luteinizing hormone (LH) and androgen concentrations, endocrine signs of polycystic ovary syndrome (PCOS). The prevalence of these abnormalities in an unselected population of adolescents is not known. We determined LH, follicle stimulating hormone (FSH), androstenedione, testosterone, dehydroepiandrosterone sulphate (DHEAS), oestradiol and prolactin concentrations in unselected population samples of adolescents with oligomenorrhoea, secondary amenorrhoea and regular menstrual cycles. A total of 2248 white, west European adolescents, aged 15.3 +/- 0.6 (mean +/- SD) years, participated. Blood was taken from 107 adolescents with regular menstrual cycles, 52 with oligomenorrhoea and four with secondary amenorrhoea. Oligomenorrhoeic adolescents had higher mean LH, androstenedione, testosterone, DHEAS and oestradiol concentrations compared with girls with regular menstrual cycles; 57% of the oligomenorrhoeic girls had LH or androgen concentrations above the 95th centile of adolescents with regular menstrual cycles. None of the 52 oligomenorrhoeic girls and only one of four girls with secondary amenorrhoea had a hypogonadotrophic endocrine pattern. The present study and available literature support the view that oligomenorrhoea in adolescents is not a stage in the physiological maturation of the hypothalamic pituitary-ovarian axis but an early sign of PCOS associated with subfertility. Physicians should consider endocrine evaluation before reassuring oligomenorrhoeic girls or prescribing oral contraceptives to these girls. PMID- 10469685 TI - A prospective, randomized comparison of ovulation induction using highly purified follicle-stimulating hormone alone and with recombinant human luteinizing hormone in in-vitro fertilization. AB - The commercial availability of highly purified, s.c. administered urinary follicle stimulating hormone (FSH) preparations for ovarian stimulation marked the beginning of a new era in the treatment of infertility. As these new formulations contain essentially no luteinizing hormone (LH), supplemental LH may be needed for optimal folliculogenesis. It was the aim of this pilot study to compare fertilization rates, embryo morphology, implantation rates and pregnancy outcomes prospectively in two age-matched patient groups: women who received highly purified FSH (FSH-HP) (n = 17), and women who received FSH-HP plus recombinant human LH (rhLH, n = 14) throughout ovarian stimulation. All patients received mid-luteal pituitary down-regulation with s.c. gonadotrophin-releasing hormone agonist (GnRHa) (leuprolide). Mean implantation rates were 26.9 and 11.9% in the FSH-HP only and FSH-HP + rhLH groups respectively. The mean clinical pregnancy/initiated cycle rate was 64.7 and 35.7% for the FSH-HP only and FSH-HP + rhLH patients respectively. FSH-HP patients and FSH-HP + rhLH patients achieved clinical pregnancy/transfer rates of 68.8 and 45.5% respectively. One patient in the FSH-HP + rhLH group had a spontaneous abortion; no pregnancy losses occurred in the FSH-HP only group. There were more cancellations for poor ovarian response among FSH-HP + rhLH patients (n = 3) than among FSH-HP patients (n = 1). The trend toward better pregnancy outcomes among patients who received FSH-HP without supplemental rhLH did not reach statistical significance. It is postulated that appropriate endogenous LH concentrations exist despite luteal GnRHa pituitary suppression, thereby obviating the need for supplemental LH administration. PMID- 10469686 TI - Recombinant human follicle stimulating hormone versus human menopausal gonadotrophin induction: effects in mature follicle endocrinology. AB - To investigate follicular effects of recombinant human follicle stimulating hormone (rhFSH) induction on women with polycystic ovary syndrome (PCOS), steroid content was compared in mature follicles obtained using a long luteinizing hormone-releasing hormone agonist plus rhFSH or human menopausal gonadotrophin (HMG) in PCOS women and controls participating in an in-vitro fertilization programme. Follicular fluids (144 samples) were collected at oocyte retrieval by individual selective aspiration. Oocyte maturity and fecundability were assessed. Plasma and intrafollicular 17beta-oestradiol, progesterone, testosterone concentrations were assayed individually. No significant difference was seen in oocyte maturity and fecundability between PCOS and controls following rhFSH, or between PCOS rhFSH and HMG group. 17beta-oestradiol, testosterone and progesterone concentrations were lower in PCOS follicular fluid following rhFSH than HMG but the difference was not significant. Progesterone concentration, 17beta-oestradiol/progesterone, 17beta-oestradiol/testosterone were significantly different between the two induction groups, for PCOS fertilized oocyte follicles (P = 0.01, P < 0.05 and P < 0.05 respectively). Steroidogenic enzymatic activity seems to be regulated in healthy follicular cells in PCOS as well as in normal patients upon ovarian induction. Following rhFSH, higher PCOS follicular progesterone concentrations leading to a theoretically increased fecundability could suggest that recombinant FSH is a better inducer which needs to be confirmed. PMID- 10469687 TI - High doses of gonadotrophin-releasing hormone antagonist in in-vitro fertilization cycles do not adversely affect the outcome of subsequent freeze thaw cycles. AB - The clinical application of gonadotrophin-releasing hormone (GnRH) antagonists instead of GnRH agonists, to prevent spontaneous premature luteinizing hormone surge during ovarian stimulation for assisted reproduction treatment has been advocated. A recent, double-blind, dose-finding study, including six dosages of the GnRH antagonist ganirelix, in women undergoing ovarian stimulation with recombinant follicle stimulating hormone (FSH), has indicated that high doses of GnRH antagonist (1 or 2 mg once daily) are associated with a low implantation rate. This follow-up study reports on the pregnancy rate after replacement of cryopreserved embryos obtained in stimulation cycles of the above-mentioned trial. Ovarian stimulation was initiated on day 2 of the cycle, with daily injections of 150 IU recombinant FSH. Ganirelix (0.0625, 0.125, 0.25, 0.5, 1.0 or 2.0 mg) was administered once daily from stimulation day 6 onwards, up to and including the day of human chorionic gonadotrophin. Retrieved oocytes were fertilized by in-vitro fertilization (IVF) or intracytoplasmic sperm injection and a maximum of three fresh embryos was transferred. Excess embryos were frozen, and subsequently used in either natural or programmed cycles. Until June 1998, 11 ongoing pregnancies (12-16 weeks after embryo transfer) were achieved from 46 cycles in which embryos had been first frozen (23.9% per transfer). Six of these 11 patients had been treated with a high dose of ganirelix (1.0 or 2.0 mg) during the IVF cycles in which the embryos were obtained. In conclusion, our data suggest that high dosages of ganirelix do not adversely affect the potential of embryos to establish clinical pregnancy in freeze-thaw cycles. PMID- 10469688 TI - Spontaneous ovarian hyperstimulation syndrome and deep vein thrombosis in pregnancy: case report. AB - This report describes a case of spontaneous ovarian hyperstimulation syndrome (OHSS) occurring in a pregnant woman carrying the factor V Leiden mutation. Even though prophylactic treatment for thrombo-embolism was adopted by administering low molecular weight heparin, the pregnancy was complicated by thromboses of the left subclavian, axillary, humeral and internal jugular veins during the second trimester of gestation. The pregnancy was managed conservatively and a healthy newborn was delivered at term. In order to avoid unnecessary laparotomy, we emphasize the importance of careful diagnosis in order to differentiate spontaneous OHSS from ovarian carcinoma, as well as the necessity to look for the presence of coagulation disorders in women affected by OHSS. PMID- 10469689 TI - Recurrent cholestasis following ovarian hyperstimulation syndrome: case report. AB - This is a case report illustrating a patient who developed recurrent cholestasis during a twin pregnancy following in-vitro fertilization (IVF) treatment. On the first occasion cholestasis developed unusually in the first trimester, and on the second occasion, it presented in the way that obstetric cholestasis (OC) is commonly seen in the third trimester. PMID- 10469690 TI - Immunoregulatory activity of decidua in spontaneous early pregnancy loss. AB - The present study aimed to address whether the immunoregulatory properties of the molecules secreted within decidua were altered in women suffering spontaneous miscarriage, compared with apparently normal fertile women. Unfractionated decidual cells from 22 women undergoing therapeutic pregnancy terminations and 25 women experiencing a sporadic spontaneous early pregnancy loss were isolated, cultured for 24 h and 72 h, and supernatants were collected. The effect of decidual supernatants on phytohaemagglutinin (PHA)-induced peripheral blood lymphocyte proliferation was investigated. Immunosuppressive activity was detected in 24 h cell culture supernatants from 91% of therapeutic abortion cases compared with only 64% of spontaneous abortion samples; 72 h supernatants from all of therapeutic abortion samples and 90% of spontaneous abortion cases suppressed lymphoproliferation. The remaining spontaneous abortion samples (36% of 24 h supernatants; 10% of 72 h supernatants) enhanced or had no effect on lymphocyte proliferation. Enhancement of lymphocyte proliferation was not observed in therapeutic abortion samples, and the association between stimulation of cell proliferation and spontaneous abortion was significant for 24 h decidual cell supernatants at 50% concentration (P = 0.02). These findings suggest that in a subgroup of women experiencing spontaneous early pregnancy loss, soluble factors within decidua display altered immune responses that may be implicated in the complex process of fetal rejection. PMID- 10469691 TI - Type and frequency of chromosome aberrations in 781 couples undergoing intracytoplasmic sperm injection. AB - Cytogenetic investigations were performed in 781 couples prior to intracytoplasmic sperm injection (ICSI) because of severe male infertility or fertilization failures in previous in-vitro fertilization attempts. Out of these 1562 patients, 1012 had a normal karyotype without any aberrations (64.8%), 204 patients had an abnormal karyotypes (13.1%). These chromosome aberrations included constitutional aberrations (4.4%), fragile sites of autosomes (3.0%), low level mosaicism of sex chromosomes (4.0%) and secondary structural chromosome aberrations (4.2%). Combinations of different types of abnormalities were stated. Another 346 patients (22.1%) showed single cell aberrations; the significance of these is unclear at the moment. Constitutional chromosome aberrations were detected in 69 patients. The following chromosome aberrations were observed: 35 sex chromosomal aberrations (comprising hyperploidies of X or Y chromosomes, mosaicisms and derivative X and Y chromosomes), 34 autosomal aberrations including 14 reciprocal translocations, five Robertsonian translocations, six inversions, one marker chromosome, one trisomy 18 mosaicism and seven other structural aberrations. Three autosomal regions showed fragile sites: 6q13 in 2.9% of the patients, 17p12 and 10q24 in 0.05% each. In conclusion, our data show that a high number of infertile couples in an ICSI programme are affected by chromosome aberrations which occur in both sexes. It is suggested that a chromosomal analysis should be performed on both partners before ICSI treatment is initiated. PMID- 10469692 TI - Factors that increase the risk of leakage during surgical removal of benign cystic teratomas. AB - The contents of mature cystic teratomas can be a potent irritant resulting in chemical peritonitis. Using a retrospective cohort, we examined the various risk factors for leakage of benign cystic teratomas during laparoscopy and laparotomy. Cyst leakage of the benign cystic teratoma contents was the primary endpoint. In all, 158 women underwent surgery for a total of 178 ovarian benign cystic teratomas. Statistical analysis was performed using chi(2), Mann-Whitney U and multivariate logistic regression analysis. A total of 115 benign cystic teratomas was successfully removed without intra-operative leakage and 63 underwent intra operative leakage either at laparoscopy or laparotomy. The likelihood of success of removing the benign cystic teratoma intact was unrelated to age, pre-operative size or surgical technique. There was no difference among cystectomies performed by laparotomy in surgeon experience or the presence of adhesions. However, surgeons with more laparoscopic experience (>35 laparoscopies/year) were less likely to have intra-operative leakage (relative risk: 0.5, 95% confidence interval: 0.2, 1.2) compared to surgeons with less experience (<20/year) at cystectomy (26.1 versus 51.2% respectively). Oophorectomy significantly reduced the frequency of intra-operative leakage at both laparoscopy and laparotomy (14.7%). These findings suggest that laparoscopic experience can reduce the risk of leakage at cystectomy. At laparotomy, lack of surgeon postgraduate years of experience was not a risk factor for leakage. PMID- 10469693 TI - The influence of donated gametes on the incidence of hypertensive disorders of pregnancy. AB - Pregnancies achieved from oocyte, sperm or embryo donation are unique, since they have resulted from donor gametes that are immunologically foreign to the mother. Thus, studying the obstetric outcome of such pregnancies may shed some light on the pathophysiology of preeclampsia, particularly in women conceiving with donated embryos, since the entire fetal genome is allogenic in these pregnancies. In this retrospective cohort study, a total of 144 women were studied. Of these, 72 were infertility patients who had conceived as a result of sperm, ovum or embryo donation and the other 72 women were age- and parity-matched control patients who became pregnant with their own gametes, either spontaneously, or following intrauterine insemination with their partner's spermatozoa. Study patients were divided into three groups depending on the origin of the donated gametes. Group 1 consisted of pregnancies achieved by intrauterine insemination with washed donor spermatozoa (n = 33). Group 2 included women who conceived using donated oocytes (n = 27) and group 3 consisted of women who conceived as a result of embryo donation (n = 12). The incidence of pregnancy-induced hypertension in the donated gametes study group was 12.5% (9/72) compared with 2.8% (2/72) in the control group. In addition, pre-eclampsia was diagnosed in 18.1% (13/72) of the donated gametes study group compared to 1.4% (1/72) in the age- and parity-matched controls. The increased incidence of gestational hypertension in pregnancies resulting from donated gametes gives evidence for a maternal genetic component, with an equally strong fetal influence, in the complicated aetiology of gestational hypertension, and pre-eclampsia in particular. PMID- 10469694 TI - Absence of human herpes virus 8 in semen from healthy Danish donors. AB - Epidemiological data indicate a sexual route of transmission of acquired immune deficiency syndrome (AIDS) associated Kaposi's sarcoma. Recently human herpes virus 8 (HHV-8) has been proposed as the aetiological agent for development of Kaposi's sarcoma. Further the virus has been reported in semen obtained from healthy men. In Denmark strict biochemical and microbiological criteria are used in combination with an intensive interview to select semen donors. Despite these strict criteria, HHV-8 may be transmitted to a recipient and even the child by the use of donor semen. We used four different polymerase chain reaction (PCR) and one nested PCR to test semen from 100 Danish donors for the presence of HHV-8 DNA. All 100 samples were consistently negative for HHV-8 DNA, while only one sample (1%) was positive for cytomegalovirus DNA. As HHV-8 was not demonstrated in any of the semen samples, we conclude that the frequency of HHV-8 in semen from Danish donors is very low. PMID- 10469695 TI - Transfundal insertion of a Veress needle in laparoscopy of obese subjects: a practical alternative. AB - Because induction of artificial pneumoperitoneum through the infra-umbilical route is associated with complications in laparoscopic procedures, especially in obese patients, we performed a prospective randomized study comparing the conventional infra-umbilical route with a transfundal route, in which the Veress needle is inserted into the peritoneal cavity through the uterine fundus. One hundred obese subjects (body mass index >/=25 kg/m(2)) scheduled for laparoscopic sterilization were randomized into two groups. In the infra-umbilical group pneumoperitoneum was achieved at a ratio (punctures/pneumoperitoneum) of 56/49 (1.14). There was one failure in this group. In the transfundal group the ratio was 53/51 (1.04). There was no clinically significant bleeding in either of the groups; nor were there any major complications. One subject in whom the infra umbilical route failed was moved to the transfundal group. This subject also underwent dilatation and curettage at the time of laparoscopy. Postoperatively she contracted chlamydial pelvic inflammatory disease. No other infections were detected postoperatively in either of the groups. In conclusion, the transfundal route of inducing artificial pneumoperitoneum proved to be easy, safe and effective. PMID- 10469696 TI - Developmental capacity of damaged spermatozoa. AB - We report the first detailed and systematic study in a mammalian system to unravel the mystery of the beginnings of life. The fertilizing ability of damaged spermatozoa at various levels of disintegration (cellular and molecular) has been investigated in homologous (mouse) and heterologous (human spermatozoon, hamster oocyte) models. Live pups were produced after destruction of spermatozoa at various cellular and molecular levels followed by injection into oocytes. We demonstrate that with damaged spermatozoa, the key point in the fertilization process is the activation of the oocyte by injection of cytosolic sperm factor. A similar fertilization rate as that using live intact spermatozoa can be achieved following activation. However, the integrity of the genetic material influenced in-vitro development of the embryos and live fetuses. This study contributes to a better understanding of the fertilizing ability of damaged spermatozoa. These findings can be applied clinically to patients with necrozoospermia or very severe oligozoospermia and in wildlife research where damaged spermatozoa from rare species can be used to regenerate young, and hence propagate the species. Also implied is the possible contribution of sperm DNA strand breakage to early pregnancy loss. PMID- 10469697 TI - Absence of testicular DAZ gene expression in idiopathic severe testiculopathies. AB - Deletions of the DAZ (deleted in azoospermia) gene family are frequently responsible for male infertility and are generally assessed by analyses of genomic DNA extracted from peripheral leukocytes. The multicopy nature of this gene prevents the distinction of intragenic deletions or deletions not involving the whole DAZ gene cluster. Thus it is still unclear whether each DAZ copy is effectively expressed in the testis. We analysed, by reverse transcription polymerase chain reaction (RT-PCR), the expression of DAZ, RBM and SRY genes, in testicular cells from infertile men affected by idiopathic severe hypospermatogenesis, obstructive azoospermia and Sertoli cell-only syndrome. Normal mRNA for DAZ, RBM and SRY were observed in obstructive azoospermia, whereas only SRY transcripts were detected when only Sertoli cells were present. Nine out of 10 patients affected by idiopathic severe hypospermatogenesis had normal expression of SRY, RBM and DAZ, while in one patient no DAZ transcript was detected, suggesting that his testiculopathy was related to the absence of DAZ expression. The lack of DAZ mRNA in testicular cells with an apparently normal DAZ gene constitution on DNA extracted from leukocytes may be explained by different hypotheses: (i) not all the copies of the DAZ gene cluster are transcribed in the germ cells and the reported patient had a small deletion involving only the active ones; (ii) the patient may be mosaic for the DAZ gene having a normal constitution in leukocytes and be deleted for DAZ gene in the testis; (iii) abnormalities of DAZ transcription may exist. These findings highlight the intrinsic interpretative difficulties of normal PCR analysis for DAZ and RBM on leukocytes and suggest caution in the use of germ cells for assisted reproductive techniques in these cases to avoid transmission of genetic abnormalities to male offspring. PMID- 10469698 TI - The outcome of intracytoplasmic sperm injection in patients with retrograde ejaculation. AB - The objective of this retrospective clinical study was to assess the benefit of assisted fertilization in cases of anejaculatory infertility due to retrograde ejaculation. We report the outcome of intracytoplasmic sperm injection (ICSI) treatment. In 16 couples in which the men suffered from retrograde ejaculation. We performed 35 cycles of ICSI with spermatozoa retrieved from post-ejaculatory urine. The patients had been instructed to alkalinize the urine by ingesting sodium bicarbonate before the procedure. The fertilization rate averaged 51.2%. Seven clinical pregnancies were achieved. Three spontaneous first trimester abortions occurred, but three live offspring were delivered and one pregnancy is ongoing. In conclusion, the use of ICSI may be feasible for patients with retrograde ejaculation who are resistant to medical treatment and whose sperm quality is so low or unpredictable that intrauterine insemination or conventional methods of in-vitro fertilization are not possible. PMID- 10469699 TI - Transurethral deroofing of midline prostatic cyst for subfertile men. AB - We evaluated the efficacy of transurethral deroofing of a midline prostatic cyst in subfertile men with one or more of the following semen abnormalities: decreased ejaculatory volume, decreased sperm motility and oligo- or azoospermia. Results from treatment of a series of 11 subfertile men with a midline prostatic cyst by transurethral deroofing of the cyst are presented. Five patients showed an improvement of seminal volume. Only one patient demonstrated an improvement of sperm count. Sperm motility was not influenced. No relationship was found between positive outcome following operation and either size of the cyst or dilatation of the seminal vesicles. Spontaneous pregnancies did not occur after transurethral deroofing of the midline prostatic cyst. In conclusion, our study suggests a poor efficacy of transurethral deroofing of a midline prostatic cyst in subfertile men with the above mentioned semen abnormalities. PMID- 10469700 TI - On-stage selection of single round spermatids using a vital, mitochondrion specific fluorescent probe MitoTracker(TM) and high resolution differential interference contrast microscopy. AB - The selection of individual round spermatids for round spermatid injection (ROSI), a prerequisite for the successful application of this infertility treatment, has been hampered by the ambiguous definition of a round spermatid and the lack of specific vital and non-vital markers. Using cells from rhesus monkey and bull, we describe a non-invasive method for the on-stage selection of individual round spermatids for ROSI, based on the polarized patterns of mitochondria, visualized in live round spermatid cells by epifluorescence microscopy after incubation with MitoTracker(TM), a vital, mitochondrion-specific fluorescent probe. The correct identification of live round spermatid was confirmed by the presence of the acrosomal granule or acrosomal cap in parallel observations by Nomarski differential interference contrast microscopy. The existence of mitochondrial polarization was first established by the labelling of MitoTracker-tagged round spermatids with spermatid-specific antibodies against proteins of nascent sperm accessory structures combined with antibodies against a nuclear pore complex component, known to disappear at the round spermatid stage. Using an inverted microscope equipped with epifluorescence, the round spermatids can be individually selected from a heterogeneous population of testicular cells labelled with MitoTracker dyes. A major advantage of this approach is that the dyes are incorporated into the paternal mitochondria, destined for rapid elimination after fertilization. In addition, the relatively high excitation and emission wavelengths of MitoTracker dyes are less harmful to DNA after their photon excitation. Before the appropriate clinical testing is conducted, the MitoTracker-based round spermatid selection may be instrumental in the training of clinical staff. PMID- 10469701 TI - Controlled comparison of conventional in-vitro fertilization and intracytoplasmic sperm injection in patients with asthenozoospermia. AB - A controlled comparison between conventional in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) has been carried out for patients with 2 embryos, 11/31 (35%) were multiple. We applied our top quality criteria to the 221 double transfers: 106 transfers with two top embryos resulted in 65 (63%) ongoing pregnancies with 37 (57%) twins, 65 transfers with one top embryo in 38 (58%) ongoing pregnancies with eight (21%) twins. In the group without top embryos, 12/52 (23%) ongoing singletons occurred, with no twins. The corresponding ongoing implantation rates were 49, 35 and 12%. This analysis suggests that single embryo transfer with an acceptable pregnancy rate might be considered if a top quality embryo is available. PMID- 10469709 TI - Effects of taurine on human embryo development in vitro. AB - Glutamine and taurine are reported to be beneficial for mouse embryo development in vitro, and we have recently shown that glutamine improves human blastocyst formation in vitro. This randomized study compared the development of supernumerary human embryos in the presence of 1 mmol/l glutamine and/or 5 mmol/l taurine from the 2-4-cell stage to the blastocyst stage. Blastocyst development and cell numbers were similar in the presence of glutamine or taurine: 52.6% and 58.3% of the embryos reached the blastocyst stage, respectively. Pyruvate uptake was similar in the presence of glutamine or taurine throughout development, as was lactate production after the 8-cell stage. Before this stage, lactate production was 4-fold higher in the presence of taurine (P < 0.001). The proportion of embryos reaching the blastocyst stage was similar with glutamine alone or with glutamine and taurine (62.5% and 47.2% respectively), as were the blastocyst cell numbers (63.0 +/- 4.6 and 61.0 +/- 5.1 respectively). In conclusion, taurine supports development of 2-4-cell human embryos to the blastocyst stage, although it does not further augment the beneficial effects of glutamine. PMID- 10469710 TI - Reconstruction of mouse oocytes by germinal vesicle transfer: maturity of host oocyte cytoplasm determines meiosis. AB - We evaluated the maturational competence of mouse oocytes reconstructed by the transfer and electrofusion of germinal vesicles (GV) into anuclear cytoplasts of GV stage oocytes (both auto- and hetero-transfers), metaphase II stage oocytes or zygotes. Following in-vitro culture, the maturation rates of the reconstructed oocytes to metaphase II did not significantly differ between auto- and hetero transfers (40/70 versus 95/144 respectively); these rates also did not differ from those of control oocytes (57/97) which were matured in vitro without micromanipulation and electrofusion. In contrast, when a GV was transferred into an enucleated metaphase II oocyte or a zygote, only a few reconstructed oocytes underwent germinal vesicle breakdown (5/30 and 2/21 respectively); moreover, none reached metaphase II stage. Cytogenetic and immunofluorescence analyses were conducted on hetero-GV oocytes that extruded a first polar body. Each oocyte showed two groups of chromosomes, one in the cytoplast and one in the polar body, as well as a bipolar spindle with twenty univalent chromosomes. Our findings suggest that oocytes reconstructed by GV transfer into a cytoplast of the same developmental stage mature normally in vitro through metaphase II. Such oocytes may be a useful research model to elucidate the cytoplasmic and nuclear mechanisms regulating meiosis and the relationships between meiotic errors and age-related changes in the oocyte. PMID- 10469711 TI - Clinical experience with ultrarapid cryopreservation of human embryos resulting from intracytoplasmic sperm injection: brief communication. AB - A total of 41 patients requested thawing of supernumerary embryos in an intracytoplasmic sperm injection (ICSI) programme. Mean patient age was 30.8 +/- 3.8 years. Embryo freezing by the ultrarapid method was performed at room temperature in 3 mol/l DMSO and 0.25 mol/l sucrose. Total freezing time was 2.5 min including filling of the straw. In the thawing process, the embryos were removed from liquid N(2), left at room temperature for 30 s, immersed for 40 s at 30 degrees C, and then successively transferred at room temperature for 10 min to each of three sucrose solutions of decreasing concentration. The embryos were kept in culture and only those that presented cleavage after 24 h were transferred. Embryos from 42 cycles were thawed and a total of 24 transfers was performed. The mean number of thawed embryos was 5.0 +/- 3.2 per cycle and the mean number of transferred embryos was 2.83 +/- 1.3. The clinical pregnancy rate per cycle obtained after the thawing process was 16. 6%. The clinical pregnancy rate per transfer was 29.2% and the implantation rate was 13.2%. The abortion rate was 14.3%. Six deliveries have been performed, with the birth of seven infants. PMID- 10469712 TI - Junctional zone contractions and embryo transfer: is it safe to use a tenaculum? AB - Applying a tenaculum to the cervix is a common practice when the correction of uterine position prior to embryo transfer is required. Our study was designed to assess junctional zone contractility before and after this procedure in 20 patients at the time of mock embryo transfer (mid-luteal phase, at commencement of down-regulation). Real-time transvaginal ultrasound and computer technology was used to evaluate the contraction pattern and frequency. When a tenaculum was applied, the total number of contractions, the number of cervico-fundal, random and opposing contractions all increased significantly (P values 0.0003, 0.005, 0. 001 and 0.007 respectively). Eleven women displayed cervico-fundal contractions, prominent opposing and random contractions were observed in all 20 patients and four patients generated fundo-cervical waves not seen in any case before stimulation with the instrument. In conclusion, manipulation with a tenaculum in the cervical area stimulates junctional zone contractions and is best avoided at the time of embryo transfer. PMID- 10469713 TI - Low-dose danazol after combined surgical and medical therapy reduces the incidence of pelvic pain in women with moderate and severe endometriosis. AB - The most effective therapy for endometriosis is a matter for debate. The aim of the present randomized study was to evaluate the efficacy of low doses of danazol on recurrence of pelvic pain in patients with moderate or severe endometriosis, who had undergone laparoscopic surgery and 6 months of gonadotrophin-releasing hormone analogue (GnRHa) therapy. After surgery, 28 patients with moderate or severe endometriosis underwent therapy for 6 months with GnRHa i. m. every 4 weeks. They were then randomized into two groups: group A (14 subjects) was treated with 100 mg/day danazol for 6 months; group B (14 subjects, control) did not receive any type of therapy. After 12 months of treatment, group A had a significantly (P < 0.01) lower pain score than group B. There was no significant difference between the groups in oestrogen concentrations, bone mineral density or side-effects. The results suggest that low-dose danazol therapy reduces recurrence of pelvic pain in patients with moderate or severe endometriosis, treated surgically, and has few or no metabolic side-effects. PMID- 10469714 TI - Preliminary results on the role of embryonic human chorionic gonadotrophin in corpus luteum rescue during early pregnancy and the relationship to abortion and ectopic pregnancy. AB - The precise mechanisms by which corpus luteum (CL) function is modulated during early pregnancy are not known. Evidence in failed pregnancies (ectopic, abortions), shows that factors other than human chorionic gonadotrophin (HCG) could be involved in its regulation. The objective of this study was to investigate the dynamics of beta-HCG, progesterone and oestradiol production in early pregnancy and its relation to embryonic quality and topographic localization. Plasma concentrations of progesterone, oestradiol and beta-HCG were studied between days +12 and +21 after an in-vitro fertilization (IVF) embryo transfer in 11 intrauterine pregnancies, 10 intrauterine abortions and seven tubal pregnancies. Tubal pregnancies and abortions were grouped according to doubling time (DT) of HCG. Results showed that oestradiol concentrations were apparently reduced in both ectopic pregnancies and abortions compared with normal pregnancies. The fall in oestradiol concentrations was seen in ectopic pregnancies with an abnormal DT for HCG and in all abortions. When the ectopic pregnancy had a normal DT, oestradiol and progesterone concentrations were normal. In abortions, the fall in oestradiol and progesterone concentrations was less influenced by the DT of HCG. These findings suggest that corpus luteum function depends on an adequate DT of HCG more than an absolute value, and with normal trophoblastic tissue the site of implantation does not affect CL function. PMID- 10469715 TI - Mannan binding lectin concentration and risk of miscarriage. AB - Recurrent miscarriage is associated with low concentrations of mannan-binding lectin (MBL), but it is not known below which value relative MBL deficiency becomes a significant risk factor. The sera of 397 patients (male and female) suffering from recurrent miscarriage and 376 controls were assayed for MBL and the data analysed. It was found that the lower the cut-off value, the greater the statistical strength of the association. It was concluded that only MBL concentrations /=28 days of oxygen in the NICU, and who were discharged from the NICU from September through November. In this subgroup, palivizumab was predicted to cost $12,000 per hospitalization averted (after taking into account savings from prevention of RSV admissions) or $33,000 per life-year saved, and the number needed to treat to avoid one hospitalization was estimated at 7.4. However, for all other subgroups, ratios ranged from $39,000 to $420,000 per hospitalization averted or $110,000 to $1,200,000 per life-year saved, and the number needed to treat extended from 15 to 152. The results were sensitive to varying assumptions about the cost and efficacy of prophylaxis, as well as the probability of hospitalization, but were less sensitive to the cost of hospitalization. CONCLUSION: In our model, the cost of prophylaxis against RSV for most subgroups of preterm infants was high relative to the benefits realized. Lower costs might permit the benefits of prophylaxis to be extended to additional groups of preterm infants. PMID- 10469765 TI - Cardiopulmonary resuscitation in the very low birth weight infant: the Vermont Oxford Network experience. AB - OBJECTIVE: The limited literature available to date suggests that the use of delivery room cardiopulmonary resuscitation (DR-CPR) is associated with very poor outcomes, especially for extremely low birth weight infants. We reviewed the cumulative experience of the Vermont Oxford Network to determine the actual utilization of DR-CPR and the neonatal outcomes of such infants. METHODS: A retrospective review of information available in the Vermont Oxford Network Database for the years 1994 to 1996. The data set was collected from 196 neonatal units who participate in the Network (data for infants 401 to 500 g were from 1996 only). Infants were eligible for study if they received DR-CPR defined as the administration of chest compressions and/or epinephrine in the delivery room as noted on the Vermont Oxford Network Database record. RESULTS: Information regarding survival was available for 27 707 newborns with birth weights from 501 to 1500 g, and 497 infants with birth weights from 401 to 500 g. There were 24 001 (86.6%) survivors. Overall DR-CPR was given to 9.3% of infants from 401 to 500 g and 6% of infants from 501 to 1500 g, 82.1% receiving chest compressions, and 66.7% receiving epinephrine. Survival of infants receiving DR-CPR was 23. 9% for infants of 401 to 500 g, and 63.3% for infants of 501 to 1500 g, compared with 16.7% and 87.9% for infants in these weight groups not receiving DR-CPR. Survival was greater for infants of 501 g or greater without DR-CPR compared with those who received this intervention within each 250-g birth weight subgroup. For infants of <1000 g, survival was 53.8% with DR-CPR compared with 74.9% without. Head ultrasounds were available for 95.5% of all surviving infants and 96.7% of infants who received DR-CPR. Overall, any grade of intraventricular hemorrhage (IVH) occurred more frequently in infants who received DR-CPR (38%) than in those who did not (21%). Grade 3 or 4 (severe) IVH was seen in 15.3% of infants who received DR-CPR compared with 4.9% of the infants who did not. Overall, survival without severe IVH occurred in 52.2% of DR-CPR infants compared with 81.3% of infants who did not require this intervention. CONCLUSION: The majority of very low birth weight and extremely low birth weight infants who receive DR-CPR survive, and at least half of such infants who survive do not have evidence of severe IVH. Further follow-up studies are required to determine the long-term neurodevelopmental outcome of such infants. The current study does not support the previously noted poor outcome in extremely low birth weight infants who receive DR-CPR. PMID- 10469766 TI - The changing pattern of hypernatremia in hospitalized children. AB - OBJECTIVES: Past studies have revealed that hypernatremia occurs primarily in infants with diarrheal dehydration. With improved infant feeding practices and the advent of pediatric critical care medicine, the pattern of hypernatremia in children has likely changed. The purpose of this study was to evaluate the current pattern of hypernatremia in hospitalized children. METHODS: Medical records were reviewed for 68 patients admitted to a large urban children's hospital during a 3-year period, all with a serum sodium greater than 150 mEq/L. The etiologies, predisposing factors, and morbidity and mortality associated with hypernatremia were evaluated. RESULTS: The average patient age was 3.9 years (range, 1 day to 19. 7 years), and the peak serum sodium concentration was 159 mEq/L (range, 151-184 mEq/L). Hypernatremia was hospital acquired in 60% of children. The majority of children (71%) were admitted for reasons other than hypernatremia. In 76% of the patients, inadequate fluid intake was the main cause of hypernatremia. Gastroenteritis contributed to the hypernatremia in only 20% (14 out of 68) of children. Eleven of these were infants <1 year of age with hypernatremia on admission. Eighty-eight percent of patients (60 out of 68) suffered from neurologic impairment, critical illness, chronic disease, or prematurity before developing hypernatremia. The overall mortality was 16%. Patients in whom hypernatremia was not corrected had a significantly higher mortality than those in whom hypernatremia was corrected (4 out of 8 [50%] vs 7 out of 60 [12%]). Peak serum sodium was no different for survivors than nonsurvivors. No deaths were attributable to cerebral edema caused by correction of hypernatremia. Neurologic complications related to hypernatremia occurred in 15% of patients. CONCLUSIONS: Hypernatremia occurs in children of all ages, with the vast majority having significant underlying medical problems. Hypernatremia caused by gastroenteritis in infants has become much less common than previously reported. Hypernatremia is primarily a hospital-acquired disease, produced by the failure to administer sufficient free water to patients unable to care for themselves. Failure to correct hypernatremia may result in a high mortality rate. PMID- 10469767 TI - Effect of computed tomography on patient management and costs in children with suspected appendicitis. AB - OBJECTIVE: Children evaluated in the emergency department for possible appendicitis are often admitted for observation, despite the widespread availability of accurate diagnostic studies, particularly computed tomography (CT). We sought to establish effective and efficient strategies for using CT to diagnose and manage children with possible appendicitis. DESIGN: Retrospective chart review and decision analysis. Setting. Emergency department of a large, urban tertiary care pediatric teaching hospital. PATIENTS: All patients admitted from January 1996 to August 1997 for suspected appendicitis. METHOD OF ANALYSIS: Three modeled strategies were empirically applied to the retrospective cohort of patients admitted for observation. Outcomes and costs under the modeled strategies were compared with those under current practice. The three strategies were: 1) to obtain CT scans on all patients and discharge those with normal findings; 2) to obtain CT scans and admit all patients; 3) to selectively obtain CT scans on those patients with a peripheral white blood cell count >10 000/mm(3) (10 x 10(9)/L) and admit all. The sensitivity and specificity of CT for diagnosing appendicitis were determined empirically from the data. A sensitivity analysis was performed. MAIN OUTCOME MEASURES: The number of preoperative inpatient observation days, total hospital costs, and the rates of both missed appendicitis and negative laparotomies. RESULTS: Of 609 patients hospitalized for possible appendicitis, 287 went directly to the operating room and 14 patients had known perforation and abscess. Three hundred eight children were observed and comprised the study cohort. Of the cohort, 112 (36.4%) underwent appendectomy and 26 (23.2%) of these had a normal appendix at pathology. Three patients were discharged from the hospital after observation and were subsequently readmitted with appendicitis (missed appendicitis). Among the 75 patients who had CT performed, the sensitivity and specificity of CT were both 97%. Under the current practice strategy, the cohort collectively accumulated 487 inpatient observation days and incurred a per patient cost of $5831. All three CT strategies would have reduced the total number of inpatient observation days, operations, negative laparotomies, as well as the per patient cost. The strategy of obtaining CT scans on all patients and then admitting them had the lowest rate of missed appendicitis. The additional cost of preventing each case of missed appendicitis under this strategy compared with the strategy of obtaining CT scans and sending home those with negative findings was $150,304. Even at the lowest reported sensitivity and specificity of CT in the literature, the ordering of the three strategies remained constant and continued to reduce total cost per patient. CONCLUSION: Compared with current practice, diagnostic strategies using CT could reduce costs and improve diagnosis, management, and outcomes for children with appendicitis. PMID- 10469768 TI - Reduction of unnecessary antibiotic therapy in newborn infants using interleukin 8 and C-reactive protein as markers of bacterial infections. AB - OBJECTIVE: To examine whether the determination of interleukin 8 (IL-8) and C reactive protein (CRP) in neonates with suspected nosocomial bacterial infection (NBI) is feasible and cost-effective in reducing antibiotic therapy. METHODS: Between April 1996 and May 1997, IL-8 was measured 260 times along with blood cultures, CRP, and immature-to-total-neutrophil (IT) ratio for suspected NBI in term and preterm neonates. All infants were retrospectively analyzed for NBI. Sensitivity, specificity, positive and negative predictive values, and 95% confidence intervals were calculated for IL-8, CRP, and IT ratio. Receiver operating characteristic curves were analyzed to determine optimal thresholds. Between June 1997 and June 1998, IL-8 was measured 215 times in newborn infants with suspected NBI and the decision to start antibiotic therapy was based on increased IL-8 and/or CRP values. A cost-effectiveness analysis was performed and sensitivity, specificity, and receiver-operating characteristic curves were reevaluated. RESULTS: At the first suspicion of NBI, the combination of IL-8 >/= 53 pg/mL and/or CRP >10 mg/L detected culture-proven NBI with 96% sensitivity. The combined culture-proven and clinical NBI were detected with 93% sensitivity and 80% specificity. The use of IL-8 reduced unnecessary antibiotic therapy for suspected NBI by 73% and was cost-effective when compared with initiating antibiotic therapy based on clinical signs alone or based on clinical signs and an increased IT ratio and/or CRP. CONCLUSIONS: The combination of IL-8 and/or CRP is a reliable and early test for the diagnosis of NBI in newborn infants. Using the combination of IL-8 and/or CRP to restrict antibiotic therapy to truly infected infants reduces unnecessary antibiotic therapy and is cost-effective. PMID- 10469769 TI - Increase in admission threshold explains stable asthma hospitalization rates. AB - BACKGROUND: Asthma morbidity and mortality has increased substantially in recent years, but asthma hospitalization rates among many geographic and sociodemographic groups have remained stable. Observations on asthma hospitalization rates and severity of acute episodes might provide valuable insight into the functioning of the health care system during this period of health care reform. OBJECTIVE: To analyze changes between 1991 and 1995 in childhood asthma hospitalization rates and severity of acute episodes. DESIGN AND METHODS: All 29 329 hospitalizations, including 2028 for asthma, for the 198 893 children (<19 years of age) in Monroe County (Rochester), New York, were studied during this 5-year period. Severity was determined by hospital record review on a 22% random sample. Using the worst oxygen saturation (SaO(2)) during the first 24 hours of hospitalization as the primary index of severity, episodes were categorized as mild (0 to >/=95), moderate (90 to 94), or severe (<90). RESULTS: Hospitalization rates are expressed as hospitalizations per 1000 child-years. The overall asthma hospitalization rate was 2.04 (95% confidence interval, 1.95 2.13). The overall annual asthma hospitalization rate remained relatively stable from 1991 (1.90) to 1995 (2.31), whereas the hospitalization rates for severe asthma rose 270%-from 0.57 to 1.55-during this period. Simultaneously, the hospitalization rates for mild asthma decreased from 0.26 to 0.12. As a proportion of all asthma hospitalizations between 1991 and 1995, severe episodes increased from 31.5% to 60.4%; conversely, mild episodes decreased from 14.1% to 4.7%. CONCLUSIONS: Severity increased significantly among children hospitalized for asthma while the overall asthma hospitalization rate remained stable. It seems that the health care system in this community has responded to an increase in severity of asthma by raising the severity threshold for admission. PMID- 10469770 TI - Predicting deterioration in previously healthy infants hospitalized with respiratory syncytial virus infection. AB - OBJECTIVE: To estimate the incidence of clinical deterioration leading to intensive care unit transfer in previously healthy infants with respiratory syncytial virus (RSV) infection hospitalized on a general pediatric unit and, to assess the hypothesis that history, physical examination, oximetry, and chest radiographic findings at time of presentation can accurately identify these infants. STUDY DESIGN: A virology database was used to identify and determine the disposition of all children 80 and an O(2) saturation <85% at time of presentation each had a specificity >97% for predicting subsequent deterioration. Each parameter, however, had a sensitivity 150 mmol/L compared with 36% of the NSG infants. PNS infants had a higher cumulative Na excretion at day 2 of life (10 +/- 2 mmol/kg vs 6 +/- 1 mmol/kg) but a less negative cumulative Na balance at 1 week (-10 mmol/kg vs -14 mmol/kg). CONCLUSION: PNS treatment was associated with lower estimated insensible water loss, a decreased incidence of hypernatremia, and an earlier diuresis and natriuresis in extremely low birth weight neonates. We speculate that PNS effects these changes through enhancement of epithelial cell maturation improving skin barrier function. PNS treatment may also enhance lung Na, K-ATPase activity leading to an earlier postnatal reabsorption of fetal lung fluid increasing extracellular volume expansion to help prevent hypernatremia. PMID- 10469774 TI - Use of state hospital discharge data to assess the morbidity from rotavirus diarrhea and to monitor the impact of a rotavirus immunization program: A pilot study in Connecticut. AB - OBJECTIVES: Now that rotavirus vaccines have been licensed and recommended for routine immunization of US infants, there is an urgent need for data to assess the morbidity from rotavirus diarrhea and to monitor the impact of a rotavirus immunization program. In a pilot study, we have assessed the usefulness of state hospital discharge data on diarrhea in children to provide this information by examining data from Connecticut. DESIGN: Retrospective analysis of discharge records from acute care, nongovernmental hospitals in Connecticut. Patients. Children 1 month through 4 years of age with a diarrhea-associated diagnosis listed on the discharge record. Setting. Connecticut, 1987 through 1996. RESULTS: During the 10-year study period, a total of 11 324 diarrhea-associated hospitalizations (49.4 hospitalizations per 10,000 children) were reported. Diarrhea-associated hospitalizations peaked during February through April, especially among children 4 to 35 months of age. The seasonality and age distribution of diarrhea-associated hospitalizations of presumed noninfectious and viral etiologies resembled those of rotavirus-associated hospitalizations. During 1993 to 1996, rotavirus was coded for 10.4% of diarrhea-associated hospitalizations increasing from 8.6% in 1993 to 14.7% in 1996. The unadjusted median cost of a diarrhea-associated hospitalization during 1987 to 1996 and 1993 to 1996 was $1,941 and $2,428, respectively. CONCLUSIONS: Diarrhea causes substantial morbidity in children from Connecticut. The winter seasonal peak of diarrhea-associated hospitalizations in children 4 to 35 months of age coinciding with the peak of rotavirus-specific hospitalizations suggests that rotavirus is an important contributor to the overall morbidity. Although our findings suggest incomplete coding of rotavirus cases, state hospital discharge data should provide sensitive and timely information to monitor the impact of a rotavirus immunization program in Connecticut. PMID- 10469775 TI - Day care attendance in the first year of life and illnesses of the upper and lower respiratory tract in children with a familial history of atopy. AB - OBJECTIVE: To examine the relationship between day care attendance and illnesses of the upper and lower respiratory tract in the first year of life. STUDY DESIGN: Prospective birth cohort study. METHODS: Children (N = 498) who had at least 1 parent with a history of allergy or asthma were enrolled at birth and followed prospectively for the first year of life. A home visit at 2 to 3 months of age and bimonthly telephone questionnaires were used to obtain information on day care arrangements, home characteristics, respiratory symptoms, and physician diagnosed illnesses of the upper and lower respiratory tract. RESULTS: Day care attendance in the first year of life was associated with two or more doctor diagnosed ear infections (OR: 2.4; 95% CI: 1.7-3.6), three or more parental reports of runny or stuffed nose (OR: 3.2; 95% CI: 1.9-5.5), a doctor's diagnosis of sinusitis (OR: 2.2; 95% CI: 1.1-4.2), and doctor-diagnosed lower respiratory illnesses (croup, bronchitis, bronchiolitis, and pneumonia; OR: 1.6; 95% CI: 1.0 2.4). For children attending day care, exposure to pets in day care, the presence of a rug or carpet in the area where the child slept in day care, and a nonresidential setting for day care all were independent predictors of two or more doctor-diagnosed ear infections. CONCLUSIONS: The results suggest that day care increases the risk of illnesses of the upper and lower respiratory tract in the first year of life for children with a familial history of atopy. Specific environmental exposures within day care, such as the presence of pets or having a rug or carpet in the area where children sleep, may increase the risk of recurrent ear infections in the first year of life among children with familial history of atopy who attend day care. PMID- 10469776 TI - Prevention of exercise-induced bronchospasm in pediatric asthma patients: A comparison of two salmeterol powder delivery devices. AB - BACKGROUND: A powder formulation of salmeterol has been shown to prevent exercise induced bronchospasm (EIB) in asthmatic children and adults; however, the delivery device (Diskhaler; Glaxo Wellcome Inc, Research Triangle Park, NC) must be reloaded after 4 doses. A new multidose powder inhaler (Diskus) provides 60 doses of salmeterol in a blister pack presentation with a dose counter. OBJECTIVE: To evaluate the safety and efficacy of 50-microg salmeterol powder via two different delivery systems (Diskhaler and Diskus) in preventing EIB in asthmatic children. STUDY DESIGN: A randomized, double-blind, double-dummy, single-dose, placebo-controlled, three-way crossover study was conducted in 24 children 4 to 11 years of age demonstrating EIB and mild to moderate asthma. Serial forced expiratory volume in 1 second (FEV(1)) was measured before and after treadmill exercise challenges conducted at 1, 6, and 12 hours after study drug administration. Adverse events were also assessed. RESULTS: During all exercise challenges, EIB-mediated reductions in FEV(1) were minimized or prevented in patients receiving single doses of salmeterol powder compared with placebo. Single doses of salmeterol powder delivered via either system were equally effective in preventing EIB. There were no drug-related adverse events, cardiovascular, or other clinically relevant safety concerns. CONCLUSIONS: Single doses of salmeterol powder delivered by either delivery system are safe and effective in preventing EIB for >/=12 hours in asthmatic children. PMID- 10469777 TI - Crowd out: evidence from the Florida Healthy Kids Program. AB - OBJECTIVE: To determine the percentage of children who had insurance coverage in the 12 months preceding enrollment in a state-subsidized program; the percentage of parents who had access to employer-based family coverage; and the cost of the families' share of the premium per month. METHODS: We randomly selected 930 families whose children were enrolled in the Florida Healthy Kids Program for a period of between 1 and 3 months and conducted telephone interviews with them in 1998 about their children's insurance coverage before program entry and their access to employer-based family coverage. There were 653 families in the final sample. RESULTS: Only 5% of the children had employer-based coverage before program enrollment. However, 26% had access to family coverage through their employers with the family share of the premiums representing on average 13% of their incomes. Access to employer-based coverage varied significantly by family income. CONCLUSIONS: Throughout the development of the State Children's Health Insurance Program legislation, policy analysts expressed concern that families may crowd out or substitute a subsidized state plan for employer-based coverage. This substitution could result in fewer improvements in access to care and health status than were anticipated, because families are simply moving to a different form of health insurance. There is some degree of crowd out in the Healthy Kids Program. The economic burden to near-poor families to purchase employer-based coverage is significant. Some degree of substitution may need to be tolerated to ensure that children receive needed health insurance. PMID- 10469778 TI - The effect of blood transfusion protocol on retinopathy of prematurity: A prospective, randomized study. AB - OBJECTIVE: Controversy exists regarding the potential influence of anemia and blood transfusions on the rate of retinopathy of prematurity (ROP) in premature infants. A prospective, randomized, masked trial was performed to determine the influence of red blood cell transfusion protocol on ROP incidence and severity in a population of high-risk infants. METHODS: A total of 50 infants with birth weights <1251 g were divided randomly into two groups beginning on day of life 29. Group 1 (n = 24) received red cell transfusions during the 6-week study period, only if certain symptom-based guidelines were met. Group 2 (n = 26) received red cell transfusions to maintain the hematocrit level above 40% for the entire 6 weeks. Infants were monitored for ROP, growth, and associated morbidity. Serial measurements of serum glucose, lactate, ferritin, total iron-binding capacity, and iron were performed. RESULTS: ROP occurred in 83% of infants in group 1, and 73% of infants in group 2. There were no statistically significant differences in ROP severity, intraventricular hemorrhage, bronchopulmonary dysplasia, necrotizing enterocolitis, or any of the laboratory values except hemoglobin (10.8 vs 13.2 g/dL) and hematocrit (33.9% vs 41.8%) between the groups. Combining data from both groups, there was no association between hemoglobin or hematocrit ratios and ROP incidence or severity. CONCLUSIONS: A transfusion policy aimed at limiting the amount of blood given to premature infants (symptom-based) during the neonatal period does not impart a significantly different risk for ROP or other associated conditions, than does a policy in which transfusions are given more liberally for replacement purposes. PMID- 10469779 TI - For-profit versus not-for-profit dialysis care for children with end stage renal disease. AB - OBJECTIVE: Over the last 2 decades, for-profit dialysis units have become the most common providers of renal replacement therapy for adults with end stage renal disease (ESRD) and have had an increasing role in the dialysis of children. We undertook a study to determine whether dialysis facility profit status influences the choice of dialysis therapy in the pediatric population. DESIGN: Cross-sectional study of national data from the Health Care Financing Administration. SETTING: Free-standing and hospital-based outpatient dialysis facilities in the United States. PATIENTS: A total of 1568 children and adolescents (0% (1%-100%) to be a positive prediction and using parent reports as the gold standard, physician estimates of gun ownership were only 65% sensitive. Approximately half (55%) of the participating pediatricians believed that pediatricians should discuss gun safety with all families, and 98% believed that pediatricians should discuss gun safety with all gun-owning families. CONCLUSIONS: Pediatricians believe that all families with guns should receive firearm safety counseling. However, pediatricians significantly underestimate the likelihood of gun ownership by specific families. Parents who own guns indicate that they would acknowledge gun ownership if their pediatrician asked about guns in the home. Therefore, rather than relying on assumptions about whether particular patients seem likely to be gun owners, pediatricians should ask all families whether they own guns. PMID- 10469782 TI - Adherence to AAP guidelines for well-child care under managed care. American Academy of Pediatrics. AB - OBJECTIVES: To determine adherence to American Academy of Pediatrics (AAP) Recommendations for Preventive Pediatric Health Care in Monroe County, New York by individual patients and individual pediatricians under managed care practice and to compare adherence-to-recommendations rates for privately insured and publicly funded managed care patients. STUDY DESIGNS AND METHODS: Using claims data for children 0 through 18 years of age cared for by pediatricians, we compared adherence to well-child care (WCC) visit recommendations for 130 572 children enrolled in a privately insured managed care system during 1992, 1993, and 1994 to 17 586 children insured by a publicly funded, Medicaid-managed care system during 1994 and 1995. Criteria for WCC visit adherence were based on 1991 AAP guidelines of 19 office visits from birth through 18 years of age. Adherence to-recommendations rates by individual pediatricians also were determined. RESULTS: Despite complete financial coverage of WCC visits (with no co-payment or deductible charges) by both insurance systems, strict adherence to AAP guidelines for WCC visits was low. Only 46% of privately insured and 35% of publicly funded children received all the recommended visits during the study period. During the same period, 17% of privately insured and 35% of publicly funded managed care patients received no WCC. There was little difference in the rate of full WCC visit adherence by age in either system with the rates ranging in privately insured patients from 49% in infants (<2 years of age) to 47% in adolescents (12 through 18 years of age) and ranging in publicly funded patients from 36% to 34% in these two age groups, respectively. Only 2% of privately insured infants had no record of WCC compared with 29% of adolescents. This contrasted with 12% of infants and 54% of adolescents who were publicly funded. Of pediatricians, <5% achieved 100% adherence to AAP guidelines for their patients (privately insured or publicly funded). Pediatricians completed an average of 52% of the recommended visits with their publicly funded patients and 68% of the recommended visits with their privately insured patients. CONCLUSIONS: WCC visits were underutilized for children in both managed care systems. Children of parents who have low incomes presumably could benefit greatest by preventive visits, but these children were less likely to receive the recommended number of WCC visits. Finding ways to increase the number of WCC visits that all children make is a major challenge, as is conducting studies that prove their worth. PMID- 10469783 TI - Brain damage in preterm newborns: might enhancement of developmentally regulated endogenous protection open a door for prevention? AB - We present a two-component model of brain white matter damage in preterm neonates. The insult component comprises infection and hypoxia-ischemia, which are both associated with inflammation-related abnormalities in the white matter. The developmental component comprises at least three factors, ie, immaturity of the ependymal/endothelial, oligodendroglial, and endogenous protection systems. All three factors are likely contributors to an increased vulnerability of the preterm newborn's white matter. In this article, we focus on recent developments in oligodendrocyte biology that support the view of certain cytokines and growth factors as oligotrophins based on their capability to enhance oligodendrocyte development or survival. We suggest that research into networks of developmentally regulated endogenous protectors (such as oligotrophins) is necessary to broaden our perspectives in brain injury prevention in preterm newborns. PMID- 10469784 TI - Magnetoencephalographic patterns of epileptiform activity in children with regressive autism spectrum disorders. PMID- 10469785 TI - RSV immune globulin prophylaxis: is an ounce of prevention worth a pound of cure? PMID- 10469786 TI - Varicella outbreaks after vaccine licensure: should they make you chicken? AB - In 1998, 3 years after vaccine licensure, child care centers (CCC) in Los Angeles County continued to report varicella outbreaks. We investigated outbreaks at 2 CCCs to determine the cause for them, such as low vaccination coverage levels or unexpected low vaccine effectiveness. We collected information on past history of varicella, illness during the outbreak, and prior varicella vaccination among CCC attendees. We found that CCC "H" had a vaccination coverage of 87% (34/39) compared with 30% (6/20) in CCC "L." The overall attack rate was lower in CCC "H" (31%) than in "L" (61%; P value =.03). Vaccine effectiveness for any varicella was 71% in "H" and 100% in "L." Vaccinated children with varicella had milder disease than unvaccinated. In conclusion, we found varicella outbreaks in CCCs with both high and low vaccination coverage. Vaccine effectiveness was within the range predicted by the literature. Vaccination led to a lower attack rate in the highly vaccinated CCC and appeared to protect from severe disease. PMID- 10469787 TI - Committee on Native American Child Health and Committee on Infectious Diseases. Immunizations for Native American children. American Academy of Pediatrics. PMID- 10469788 TI - Joint statement of the American Academy of Pediatrics (AAP) and the United States Public Health Service (USPHS). PMID- 10469789 TI - Thimerosal in vaccines--An interim report to clinicians. American Academy of Pediatrics. Committee on Infectious Diseases and Committee on Environmental Health. PMID- 10469790 TI - Possible association of intussusception with rotavirus vaccination. American Academy of Pediatrics. Committee on Infectious Diseases. PMID- 10469792 TI - Introduction PMID- 10469791 TI - Foreword PMID- 10469793 TI - Therapeutic orphans. PMID- 10469794 TI - An update on the therapeutic orphan. PMID- 10469795 TI - Therapeutic orphans: European perspective. PMID- 10469796 TI - Is pediatric labeling really necessary? PMID- 10469797 TI - Off-label uses of drugs in children. PMID- 10469798 TI - Essential drugs for infants and children: North American perspective. PMID- 10469799 TI - Essential drugs for infants and children: European perspective. PMID- 10469800 TI - Lack of pediatric drug formulations. AB - Many drugs frequently used in infants and young children are not available in suitable dosage forms. Liquid dosage forms must be prepared extemporaneously, while using appropriate excipients. However, it is critical to determine the stability of various drugs at clinically important concentrations and practical storage conditions. It is of concern that few funding agencies are willing to support research on the development of stable liquid dosage forms for pediatric patients. The need for such data will continue, because it is unlikely that all drugs approved for adults will also be labeled simultaneously for potential use in infants and children. Presentations and publications on stable drug formulations will offer the opportunities for pediatric patients to receive the desired drugs and doses most effectively and safely. PMID- 10469801 TI - A look at the future of pediatric therapeutics: an investigator's perspective of the new pediatric rule. PMID- 10469802 TI - European regulatory authorities and pediatric labeling. PMID- 10469803 TI - Patient recruitment: US perspective. AB - There are many ethical, legal, economic, scientific, and practical problems associated with conducting drug trials in children. The single most difficult is subject identification and enrollment (ie, recruitment). This article reviews various aspects of the recruitment process and proposes potential solutions to recruitment problems. PMID- 10469804 TI - Patient recruitment--European perspective. AB - A considerable number of patients have to be recruited in a clinical trial to obtain solid results. In pediatric studies, patient recruitment is frequently problematic. In the simple common childhood illnesses, the number of recruitable patients is certainly large, but they may be hard to reach, and the imbalance between potential benefit and inconvenience of participation may reduce motivation to enroll. In severe diseases, the balance may be right, but the available number of patients may be small. Good communication with the child and family, as well as the motivation of colleagues to admit, is another key element in success. Proper study design, including realistically identified sources of patients, reasonable inclusion, and exclusion criteria, also are required. PMID- 10469805 TI - Optimal sampling theory: An overview of its application to pharmacokinetic studies in infants and children. PMID- 10469806 TI - Stable isotopes labeling of drugs in pediatric clinical pharmacology. PMID- 10469807 TI - Phenotyping of drug metabolism in infants and children: potentials and problems. PMID- 10469808 TI - The Pediatric Pharmacology Research Unit (PPRU) Network and its role in meeting pediatric labeling needs. PMID- 10469809 TI - Current incidence of retinopathy of prematurity, 1989-1997. AB - OBJECTIVE: To report the current incidence and the need for surgery for retinopathy of prematurity (ROP) in neonates (22-36 weeks' gestational age [GA], July 1, 1989 through June 30, 1997). STUDY DESIGN: Retrospective analyses using computerized perinatal database kept on all admissions, a review of patient charts, and eye examination log books. SETTING: Level 3 regional referral NICU. PATIENTS: A total of 2528 infants <37 weeks' GA were admitted during this time. Of these infants, 950 met the criteria for eye examination beginning at 4 to 6 weeks of age and repeated every 2 weeks until complete vascularization of the retina or death or discharge. RESULTS: The incidence of ROP was (202/950) 21.3% for any stage and 4.6% (44/950) for stage 3 ROP or greater. No ROP was noted in infants born at >32 weeks' GA. No infant born at >28 weeks needed retinal surgery. Using birth weight (BW) criteria, stage 3 ROP was not noted in infants with BWs >1500 g; retinal surgery was not needed in infants with BWs >1000 g. A number of perinatal factors were associated with ROP on univariate analysis. However, using multiple logistic regression analyses of these factors, only GA and days on supplemental oxygen therapy were associated significantly with the development of ROP. Despite increased survival of extremely low BW infants, we found a considerable reduction in incidence and severity of ROP compared with reports from an earlier chronological period. However, infants <28 weeks' GA or with BWs <1000 g were still at considerable risk for retinal surgical treatment for ROP. CONCLUSION: We conclude that the incidence and severity of ROP have decreased significantly in the present era of surfactant therapy. PMID- 10469810 TI - Television-viewing habits and sleep disturbance in school children. AB - OBJECTIVE: To investigate the relationship between specific television-viewing habits and both sleep habits and sleep disturbances in school children. METHODS: The parents of 495 children in grades kindergarten through fourth grade in three public elementary schools completed two retrospective survey questionnaires, one assessing their children's sleep behaviors and the other examining television viewing habits of both the child and the family. Sleep domains assessed included bedtime resistance, sleep onset delay, sleep duration, anxiety around sleep, parasomnias, night wakings, and daytime sleepiness. Teachers from all three schools also completed daytime sleepiness questionnaires (N = 402) for the sample. RESULTS: Most of the television-viewing practices examined in this study were associated with at least one type of sleep disturbance. Despite overall close monitoring of television-viewing habits, one quarter of the parents reported the presence of a television set in the child's bedroom. The television viewing habits associated most significantly with sleep disturbance were increased daily television viewing amounts and increased television viewing at bedtime, especially in the context of having a television set in the child's bedroom. The sleep domains that appeared to be affected most consistently by television were bedtime resistance, sleep onset delay, and anxiety around sleep, followed by shortened sleep duration. The parent's threshold for defining "problem sleep behavior" in their child was also important in determining the significance of the association between sleep disturbance and television-viewing habits. CONCLUSION: Health care practitioners should be aware of the potential negative impact of television viewing at bedtime. Parents should be questioned about their children's television-viewing habits as part of general screening for sleep disturbances and as part of anticipatory guidance in regards to healthy sleep habits in children. In particular, the presence of a television set in the child's bedroom may be a relatively underrecognized, but important, contributor to sleep problems in school children. PMID- 10469811 TI - Health system factors contributing to breastfeeding success. AB - OBJECTIVE: To identify and characterize health care system factors that contribute to successful breastfeeding in the early postpartum period. STUDY DESIGN: A prospective 8-week cohort study of 522 women at five area hospitals who had a vaginal delivery of a healthy, full-term single child and who intended to breastfeed. Mothers and infants had free access to each other for breastfeeding during the hospital stay. Data were obtained through chart review and surveys. In person postpartum interviews in the hospital and 4- and 8-week telephone interviews were used to determine participants' perceptions of breastfeeding support by hospital personnel, home visit nurses, and family and friends. The hospital in-person interview with each mother was conducted before discharge to confirm maternal interest and intent to breastfeed. Questions were asked regarding breastfeeding information and support provided by medical and nursing personnel. Mothers were asked to rate the quality of information, as well as the degree of support they received for breastfeeding. Mothers also were asked to rate their hospital breastfeeding experience. A second interview was conducted by telephone 4 weeks after birth. The focus of this interview was to ascertain the rating of their breastfeeding experience, the quality of their interactions with health care professionals, and whether supplemental formula was being provided to the infant. If supplemental formula was being provided, the mothers were asked to quantify the volume and frequency of supplementation. A final telephone interview was conducted when the infants were 8 weeks of age. This interview determined the continuance or cessation of breastfeeding and information about formula supplementation, as in the 4-week interview. Mothers were given a journal and asked to note all telephone calls, clinic visits, and home nurse visits that related to breastfeeding issues and concerns. Demographic data examined included maternal age, marital status, highest level of education reached, race, employment, insurance coverage, and length of stay in the hospital. Pregnancy characteristics included prenatal care, parity, and gravity. Infant characteristics included gestational age and birth weight. Other factors examined included maternal rating of the support received from the infant's father for the decision to breastfeed, the time the infant spent in the mother's hospital room, and whether the infant was breastfed in the delivery room. RESULTS: The women were mostly white (90%), educated (82% had some college education), married, older (mean maternal age of 29.3 years), and insured (92% commercial). The primary outcome of interest was success at breastfeeding. Success was determined based on each mother's initial estimate of the planned duration of breastfeeding. Of the participants, 76% breastfed successfully for at least as long as they had initially planned. Seventeen percent of the mothers had stopped breastfeeding at the time of the 4-week interview, and 29% had stopped by the 8-week interview. Of the infants' fathers, 97% were reported by the mothers to be supportive of the decision to breastfeed. Once discharged, 98% of mothers expected to have help with the household chores. Eighty percent rated their hospital breastfeeding experience as good or very good. However, only 56% rated hospital breastfeeding support as good or very good, and only 44% spoke with a lactation consultant while in the hospital. Of those who spoke with the lactation consultant, 85% felt more confident afterward. Hospital nurses talked with 82% of women, and 97% of these found this helpful. Seventy-four percent reported receiving a home nursing visit after discharge, and of these, 82% found it helpful. Successful mothers were significantly more likely to report that the visiting nurse watched them breastfeed and asked how it was going. Mothers were more likely to call or visit family and friends with concerns about breastfeeding than PMID- 10469813 TI - Measurement of blood pressure in children: recommendations and perceptions on cuff selection. AB - OBJECTIVE: . To review some of the recent recommendations on blood pressure (BP) cuff selection, including those by the Task Force and its update, and to survey the perceptions on cuff selection among practitioners. METHODS: The study was conducted in two parts. In the first, we selected three brands of commonly used infant, child/pediatric, small adult, adult, and large adult BP cuffs. From the median width of each, we derived the required upper arm length (UAL) for the cuff to cover three quarters or two thirds of the UAL, and with the help of a published normal UAL at various ages, we matched the derived UAL with the corresponding age at the 50th percentile. Similarly, we derived the midupper arm circumference (UAC) so that the available cuffs would cover 40% of the UAC, and by using the published normal UAC at various ages, we matched the derived UAC with the corresponding age at the 50th percentile. The second part of the study involved a survey by multiple choice questionnaire mailed to 400 hospital- and office-based pediatricians, residents, and nurses at the Children's Hospital of Michigan. Included in the survey were questions about the age at which practitioners would choose the cuffs described above; the minimum age they would consider using an adult cuff in a pediatric patient with an average height and weight; selecting a cuff using UAL as a criterion; selecting a large versus a small cuff when the appropriate cuff size is not available; and the Task Force definition of hypertension. RESULTS: Using three quarters of the UAL as a criterion, it seems that a large adult cuff should be appropriate for an average size 6-year-old child and that using two thirds of UAL as a criterion, the cuff should be appropriate for an average-size 7- to 8-year-old child. Similarly, by using 40% of the mid-UAC as a criterion, an adult cuff would be of no use in an average pediatric patient at any age. Our survey revealed that 57% of practitioners would consider using a neonatal cuff for patients up to 1 month of age, 65% would use an infant cuff for those 1 year of age, 49% would use a child/pediatric cuff for those 5 years of age, and 84% would use a small adult cuff for those 10 years of age and older. Most (83%) of the practitioners would consider using an adult cuff in children 11 years of age and older. Practitioners are likely to use a smaller cuff than is appropriate by two thirds or three quarters of UAL criteria, and a larger cuff than is appropriate, particularly in older children, by 40% of UAC criteria. Using UAL as a criterion, a majority (59%) of practitioners use cuffs that cover two thirds of the distance between the axilla and the cubital fossa. Ninety-two percent of practitioners believe that a smaller cuff causes a moderate to significant increase in the BP reading, and 55% believe that a larger cuff causes a similar decrease in the BP reading. A significant number of practitioners (44%) did not know the Task Force definition of hypertension, including 42% of attendings, 44% of residents, and 50% of nurses. CONCLUSIONS: The Task Force and the Working Group recommendations on BP cuff selection need to be reviewed. A new multicenter study, using uniform criteria for cuff selection, may be necessary to establish the accuracy of the published nomogram on normal BP in children. More awareness is required on part of practitioners of the current recommendations on BP measurement and the definition of hypertension. Finally, the labeling of BP cuffs as infant, pediatric, small adult, adult, and large adult is misleading, and such designations should be eliminated. Cuff sizes should be standardized, indicate bladder size, and be uniformly color-coded for convenience. PMID- 10469812 TI - Oral rehydration solution for acute diarrhea prevents subsequent unscheduled follow-up visits. AB - BACKGROUND: Oral rehydration solutions (ORS) for the treatment of acute diarrhea remain an underutilized therapy in the United States, despite multiple clinical trials confirming their efficacy and safety. Economic barriers to their use have been identified. OBJECTIVE: To determine whether providing ORS to patients at the time of their office visit for acute diarrhea can increase ORS utilization and reduce unscheduled follow-up visits. DESIGN: Randomized, controlled clinical trial. SETTING: Seven health centers of a large health maintenance organization. PARTICIPANTS: Children (N = 479) 0 to 60 months of age with acute diarrhea (at least three watery or loose stools in the previous 24 hours for /=90th percentile) with those who were born appropriate for-gestational age (10th-89th percentile) to determine whether there are differences in growth and fatness in early childhood associated with birth weight status. DESIGN AND METHODS: National sample of 3192 US-born non-Hispanic white, non-Hispanic black, and Mexican-American children 3 to 6 years of age (36-83 months) examined in the third National Health and Nutrition Examination Survey and for whom birth certificates were obtained. On the birth certificates, length of gestation from the mother's last menstrual period was examined for completeness, validity, and whether the pattern of missing (n = 141) and invalid data (n = 147) on gestation was random. Gestation was considered invalid when >44 weeks, or when at gestations of 3' delineation of transcription units (and promoters) still requires additional experiments. Similarly, functional predictions made with reference to previously characterized homologues are leaving >50% of human genes unannotated or classified in uninformative categories ('kinase', 'ATP-binding', etc.). In the context of functional genomics, large-scale gene expression studies using massive cDNA tag sequencing, two-dimensional gel proteome analysis or microarray technologies are the only approaches providing genome-scale experimental information at a pace consistent with the progress of sequencing. Given the difficulty and cost of characterizing genes one by one, academic and industrial researchers are increasingly relying on those methods to prioritize their studies and choose their targets. The study of expression patterns can also provide some insight into the function, reveal regulatory pathways, indicate side effects of drugs or serve as a diagnostic tool. In this article, I review the theoretical and computational approaches used to: (i) identify genes differentially expressed (across cell types, developmental stages, pathological conditions, etc.); (ii) identify genes expressed in a coordinated manner across a set of conditions; and (iii) delineate clusters of genes sharing coherent expression features, eventually defining global biological pathways. PMID- 10469834 TI - Breakthroughs in molecular and cellular mechanisms underlying X-linked mental retardation. AB - Although genetic causes of X-linked mental retardation (XLMR) are heterogeneous and complex, recent concerted actions between physicians and biologists have allowed some major difficulties to be overcome and led to the identification of an increasing number of genes involved in these conditions. Indeed, over the past 2 years significant progress has been made in understanding the molecular basis underlying not only XLMR, where there are distinguishing phenotypic or genetic markers (syndromal forms of XLMR), but also non-specific (or idiopathic) mental retardation (MRX). Recent breakthroughs have shown that genes responsible for these latter conditions encode for proteins involved in signalling pathways which regulate cytoskeleton organization, synaptic vesicle transport and, maybe, other cellular functions. Also, they suggest a provacative picture that conceptualizes MRX as disorders resulting from a dysfunctioning of genes required for processes such as the remodelling, establishment and stabilization of connections between neuronal cells. Such processes are crucial for the development of intellectual and cognitive functions. As these functions begin to evolve mainly in post-natal stages through contact with diverse stimuli and environments, a potential therapeutic approach would be the development of drugs that target cellular signalling pathways shown to be implicated in MRX. PMID- 10469836 TI - Heart to heart: from nuclear proteins to Emery-Dreifuss muscular dystrophy. AB - Emery-Dreifuss muscular dystrophy has some remarkably specific features, with only cardiac and skeletal tissues being affected. Equally remarkably, the disease is caused by mutations in widely expressed genes for the nuclear membrane/lamina proteins, emerin and lamin A/C. How do mutations in proteins at the heart of the cell lead to stiff joints and sudden heart failure? This and related questions are the subject of this review. PMID- 10469835 TI - Osteopetrosis and osteoporosis: two sides of the same coin. AB - Together, osteoporosis and osteopetrosis comprise a substantial proportion of the bone diseases that severely affect humans. In order to understand and effectively treat these disorders, an understanding of the mechanisms controlling bone remodelling is essential. While numerous animal models of bone disease have been generated, the lack of correlation between these animal models and human disease has limited their utility in terms of defining therapeutic strategies. The generation and analysis of cathepsin K knockout mice has resulted in a model for pycnodysostosis, a rare human osteopetrotic disease, and is now providing considerable insights into both osteoclast function and potential therapeutic strategies for the treatment of bone disease. This review highlights the importance of genes such as cathepsin K in understanding bone remodelling and illustrates a new trend towards understanding bone disease as a complete entity rather than as a series of unrelated disorders. PMID- 10469837 TI - The many faces and factors of orofacial clefts. AB - Orofacial clefts are congenital structural anomalies of the lip and/or palate that affect approximately 1/1000 live births. Their frequent occurrence as well as their extensive psychological, surgical, speech and dental involvement emphasize the importance of understanding the underlying causes. The etiology of orofacial clefts is complex, including multiple genetic and environmental factors. Rare forms, where they occur as one component of multiple congenital anomaly syndromes, have Mendelian or teratogenic origins; the non-syndromic forms of orofacial clefts are more common and are likely due to secondary gene environment interactions. Recent advances in both molecular and quantitative approaches have begun to identify the genes responsible for the rare syndromic forms of cleft and have also identified both candidate genes and loci for the more common and complex non-syndromic variants. Animal models, in particular the mouse, have also contributed greatly to an understanding of these disorders. This review describes genes that are involved in orofacial clefts in humans and animal models and explores genetic approaches to identifying additional genes and gene environment interactions that constitute the many factors of orofacial clefts. PMID- 10469838 TI - Autosomal dominant polycystic kidney disease: clues to pathogenesis. AB - Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutation of one of two genes: PKD1 (16p13.3) or PKD2 (4q13-23). PKD1 accounts for approximately 85% of pedigrees and is associated with significantly more severe cystic disease. The ADPKD genes encode proteins, polycystin-1 and polycystin-2, which are very different in size and structure, but which have a region of homology and may interact as part of the same complex. Polycystin-1 is a large, integral membrane protein ( approximately 460 kDa) predicted to be involved in cell-cell and/or cell-matrix interactions. Polycystin-2 ( approximately 110 kDa) is related to polycystin-1 and voltage-activated and transient receptor potential channel subunits, suggesting that the polycystins may also be associated with ion transport. A polycystin complex could regulate cellular events (that are abnormal in ADPKD) in response to specific extracellular cues, mediated by controlling cellular Ca(2+)levels and/or other signalling pathways. Recently, two further polycystin-like molecules have been identified, indicating roles for this novel protein family beyond the kidney. A wide range of different mutations to the PKD1 or PKD2 gene have been detected, most predicted to truncate and inactivate the proteins. A somatic second hit may be required for focal cyst development, although there is widespread immunohistochemical evidence of polycystin expression in cystic epithelia. Disruption of the mouse Pkd1 gene leads to death in the perinatal period with massive cystic expansion in homozygotes and age related cyst development in heterozygotes. Normal renal development in Pkd1(del34/del34)mice up to embryonic day approximately 15.5 suggests a role for polycystin-1 in developing and maintaining the tubular architecture, consistent with the localization of the protein, rather than nephron induction. Renal cystic disease in homo- and heterozygotes of a Pkd2 mouse model with a disrupted exon 1 inserted in tandem with the normal exon (and prone to somatic recombination, which inactivates the gene) supports a role for somatic events in cystogenesis. PMID- 10469839 TI - Towards a molecular understanding of Prader-Willi and Angelman syndromes. AB - Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are two distinct neurological disorders that map to human chromosome 15q11-q13 and involve perturbations of imprinted gene expression. PWS is caused by a deficiency of paternal gene expression and AS is caused by a deficiency of maternal gene expression. Experiments in the last year have focused on molecular analysis of the human chromosomal region as well as the homologous region on central mouse chromosome 7. New transcripts and exons have been identified and the epigenetic status of the PWS/AS region in mice and humans has been examined. The imprinting center that is hypothesized to control the switch between the maternal and paternal epigenotypes has also been characterized in greater detail and a mouse model that deletes the homologous element demonstrates a conservation in imprinting center function between mice and humans. In addition, analysis of non deletion AS patients has revealed that UBE3A intragenic mutations are found in a significant number of cases. However, both human patients and mouse model systems indicate that other genes may also contribute to the AS phenotype. Thus, although much has been learned in the last year, considerable information is still required before these complex syndromes are fully understood. PMID- 10469840 TI - The limb-girdle muscular dystrophies-multiple genes, multiple mechanisms. AB - In the field of muscular dystrophy, advances in understanding the molecular basis of the various disorders in this group have been rapidly translated into readily applicable diagnostic tests, allowing the provision of more accurate prognostic and genetic counselling. The limb-girdle muscular dystrophies (LGMD) have recently undergone a major reclassification according to their genetic basis. Currently 13 different types can be recognized. Amongst this group, increasing diversity of the mechanisms involved in producing a muscular dystrophy phenotype is emerging. Recent insights into the involvement of the dystrophin glycoprotein complex in muscular dystrophy suggests that its members may play distinct or even multiple roles in the maintenance of muscle fibre integrity. In other forms of LGMD, proteins have been implicated which may be important in intracellular signalling, vesicle trafficking or the control of transcription. As these various mechanisms are more fully elucidated, further insights will be gained into the pathophysiology of muscular dystrophy. At a practical level, despite the marked heterogeneity of this group real progress can at last be made in determining a precise diagnosis. PMID- 10469841 TI - A family of mammalian anion transporters and their involvement in human genetic diseases. AB - Tremendous advances in human genetics have been made in recent years, as the fruits of the Human Genome Project are facilitating the identification of genes associated with myriad genetic diseases. Among the many triumphs in positional (and positional candidate) cloning are a number of cases where apparently unrelated diseases have been found to share common genetic origins. A vivid example of this has unfolded in the past few years with the identification of the genes causing diastrophic dysplasia, congenital chloride diarrhoea and Pendred syndrome. While these three disorders are clinically distinct, the associated genes ( DTDST, CLD and PDS, respectively) emanate from a well conserved family of genes that all encode anion transporters. Our current knowledge of these diseases coupled with new insights about the implicated genes and proteins illustrates the complex nature of mammalian genomes, especially with respect to the evolutionary subtleties of protein families and tissue-specific gene expression. PMID- 10469842 TI - Nonsense-mediated mRNA decay in health and disease. AB - All eukaryotes possess the ability to detect and degrade transcripts harboring premature signals for the termination of translation. Despite the ubiquitous nature of nonsense-mediated mRNA decay (NMD) and its demonstrated role in the modulation of phenotypes resulting from selected nonsense alleles, very little is known regarding its basic mechanism or the selective pressure for complete evolutionary conservation of this function. This review will present the current models of NMD that have been generated during the study of model organisms and mammalian cells. The physiological burden of nonsense transcripts and the emerging view that NMD plays a broad and critical role in the regulation of gene expression will also be discussed. Such issues are relevant to the proposal that pharmacological manipulation of NMD will find therapeutic application. PMID- 10469843 TI - The genetics of disorders with synuclein pathology and parkinsonism. AB - Despite being considered the archetypal non-genetic neurological disorder, genetic analysis of Parkinson's disease has shown that there are at least three genetic loci. Furthermore, these analyses have suggested that the phenotype of the pathogenic loci is wider than simple Parkinson's disease and may include Lewy body dementia and some forms of essential tremor. Identification of alpha synuclein as the first of the loci involved in Parkinson's disease and the identification of this protein in pathological deposits in other disorders has led to the suggestion that it may share pathogenic mechanisms with multiple system atrophy, Alzheimer's disease and prion disease and that these mechanisms are related to a synuclein pathway to cell death. Finally, genetic analysis of the synuclein diseases and the tau diseases may indicate that this synuclein pathway is an alternative to the tau pathway to cell death. PMID- 10469844 TI - Ataxic mouse mutants and molecular mechanisms of absence epilepsy. AB - Mouse genetic models for common human diseases have been studied for most of the 20th century. Although many polygenic strain differences and spontaneous single gene mutants have been extensively characterized over the years, knowing their innermost secrets ultimately requires the identity of the mutated genes. One group of neurological mutants, detected initially due to cerebellar dysfunction, was identified as models for epilepsy when they were unexpectedly found to have spike-wave seizures associated with behavioral arrest, a central feature of absence or petit-mal epilepsy. A further surprise was that recently identified defective genes encode different subunits of voltage-gated Ca(2+)channels (VGCCs), implying common seizure mechanisms. In this review we first consider these spontaneous mutants with VGCC defects in the context of other mouse models for epilepsy. Then, from the new wave of genetic and functional studies of these mutants we discuss their prospects for yielding insight into the molecular mechanisms of epilepsy. PMID- 10469845 TI - Molecular genetics of the Finnish disease heritage. AB - Finland, located at the edge of the inhabitable world, is one of the best-studied genetic isolates. The characteristic features of population isolates-founder effect, genetic drift and isolation-have, over the centuries, shaped the gene pool of the Finns. Finnish diseases have been a target of extensive genetic research and the majority of some 35 disease genes enriched in this population have been identified; the molecular and cellular consequences of disease mutations are currently being characterized. Special strategies taking advantage of linkage disequilibrium have been efficiently used in the initial mapping and restriction of Finnish disease loci and this has stimulated development of novel statistical approaches in the disease gene hunt. Identification of mutated genes has provided tools for detailed analyses of molecular pathogenesis in Finnish diseases, many of which reveal a distinct tissue specificity of clinical phenotype. Often these studies have not only clarified the molecular detail of Finnish diseases, but also provided novel information on biological processes and metabolic pathways essential for normal development and function of human cells and tissues. PMID- 10469846 TI - Forward genetics in mammalian cells: functional approaches to gene discovery. AB - Definitive proof of function in biological systems requires genetic analysis. Only when the loss of a particular protein corresponds to the loss of a specific function can one be sure that the protein truly affects the function. Changing the pattern of gene expression through random mutagenesis or by introducing expression libraries, followed by selection of mutant or variant cells and identification of a missing or overexpressed protein, has the power to reveal or confirm the roles of specific components of signaling pathways and to provide mutant cell lines and cDNA reagents to be used in defining detailed mechanisms through structure-function analyses. These examples of forward genetics contrast with reverse genetic approaches, where the function of a known gene product is explored by knockout or replacement. Here we review a broad range of techniques that have been used to alter gene expression randomly in mammalian cells, with examples of specific discoveries that have resulted from these applications of forward genetics. PMID- 10469847 TI - Genetics of HIV-1 infection: chemokine receptor CCR5 polymorphism and its consequences. AB - The chemokine receptor gene, CCR5, has become a central theme in studies of host genetic effects on HIV-1 pathogenesis ever since the discovery that the CCR5 molecule serves as a major cell surface co-receptor for the virus. A growing number of genetic variants within the coding and 5' regulatory region of CCR5 have been identified, several of which have functional consequences for HIV-1 pathogenesis. Here we review the CCR5 literature describing CCR5 polymorphism and the functional ramifications that several of these variants have on HIV-1 infection and progression to AIDS. The multiplicity of CCR5 genetic effects on HIV-1 disease underscores the critical importance of this gene in controlling AIDS pathogenesis and provides the logic for develop-ment of therapeutic strategies that target the interaction of HIV-1 envelope and CCR5 in HIV-1 associated disease. PMID- 10469848 TI - Williams-Beuren syndrome: genes and mechanisms. AB - Williams-Beuren syndrome (WBS; OMIM 194050) is caused by heterozygous deletions of approximately 1.6 Mb of chromosomal sub-band 7q11.23. The deletions are rather uniform in size as they arise spontaneously by inter- or intrachromosomal crossover events within misaligned duplicated regions of high sequence identity that flank the typical deletion. This review will discuss the status of the molecular characterization of the deletion and flanking regions, the genes identified in the deletion region and their possible roles in generating the complex multi-system clinical phenotype. PMID- 10469849 TI - Mouse ENU mutagenesis. AB - The progress of human genome sequencing is driving genetic approaches to define gene function. Strategies such as gene traps and chemical mutagenesis will soon generate a large mutant mouse resource. Point mutations induced by N -ethyl- N nitrosourea (ENU) provide a unique mutant resource because they: (i) reflect the consequences of single gene change independent of position effects; (ii) provide a fine-structure dissection of protein function; (iii) display a range of mutant effects from complete or partial loss of function to exaggerated function; and (iv) discover gene functions in an unbiased manner. Phenotype-driven ENU screens in the mouse are emphasizing relevance to human clinical disease by targeting cardiology, physiology, neurology, immunity, hematopoiesis and mammalian development. Such approaches are extremely powerful in understanding complex human diseases and traits: the base-pair changes may accurately model base changes found in human diseases, and subtle mutant alleles in a standard genetic background provide the ability to analyze the consequences of compound genotypes. Ongoing mouse ENU mutagenesis experiments are generating a treasure trove of new mutations to allow an in-depth study of a single gene, a chromosomal region or a biological system. PMID- 10469850 TI - Idiopathic edema. AB - Idiopathic edema is a syndrome of real or perceived excessive weight gain. This article reviews what is known about the possible causes, evaluation, and treatment. Although the cause is unknown but often thought to be due to secondary hyperaldosteronism, primary abnormalities of the hypothalamus, thyroid, dopaminergic release or renal dopaminergic metabolism, vascular basement membrane, or capillary sphincter control could perhaps contribute in some patients. The diagnosis requires careful attention to possible abnormalities of the liver, heart, kidneys, gastrointestinal tract, thyroid, and pancreas. The history must include an evaluation for risks of bulimia and purging; diuretic and laxative screening should be performed. Specific records of daily weights, urinary outputs, and menstral cycle dates are useful. Treatment may include dietary counseling to provide weight control and a constant carbohydrate intake, treatments for depression, compression stockings, spironolactone, amiloride, angiotensin II inhibitors, or sympathomimetic agents, depending on the severity and timing of the patient's symptoms. Unfortunately, idiopathic edema may be a multifactorial disorder that has not been completely delineated. Further research into possible causative mechanisms is required before a more useful algorithm for evaluation and treatment is available. PMID- 10469851 TI - Comparing continuous hemofiltration with hemodialysis in patients with severe acute renal failure. AB - Continuous venovenous hemofiltration (CVVH) or CVVH with additional diffusive dialysis (CVVH-D) has theoretical advantages in treating severe acute renal failure (ARF), but no prospective clinical trials or restrospective comparison studies have clearly shown its superiority over intermittent hemodialysis (HD). To evaluate this question, all 349 adult patients with ARF receiving renal replacement therapy (RRT) at our medical center during 1995 and 1996 were analyzed using multivariate Cox proportional hazards methods. Initial univariate analysis showed the odds of death when receiving initial CVVH to be more than twice those when receiving initial HD (risk for death, 2.03; P < 0.01). Progressive exclusion of patients in whom the RRT modality might not be open to choice and the risk for death was very high (systolic blood pressure < 90 mm Hg; total bilirubin level > 15 mg/dL; or total RRT < 48 hours) for total RRT left 227 patients in whom the risk for death was 1.09 (95% confidence interval [CI], 0.67 to 1.80; P = 0.72) for initial CVVH, virtually equivalent to the risk for initial HD. Comorbid indicators significantly associated with death or failure to recover renal function included: older age; medical rather than surgical diagnosis; preexisting infection or trauma and liver disease as primary diagnoses; and abnormal bilirubin level or vital signs at initiation of RRT. These results show that the high crude mortality rate of patients undergoing CVVH was related to severity of illness and not the treatment choice itself. With the addition of more inclusive comorbidity data and a broader spectrum of interim outcomes, this type of analysis is a practical alternative to what would almost assuredly be a cumbersome and costly prospective, controlled trial comparing traditional HD with CVVH. PMID- 10469852 TI - Increased genomic damage in lymphocytes of patients before and after long-term maintenance hemodialysis therapy. AB - This study investigates spontaneous genomic damage in peripheral lymphocytes of 19 patients with severe end-stage renal disease not enrolled onto a maintenance hemodialysis (MHD) program (creatinine level, 5.4 to 10.5 mg/dL) and 16 long-term MHD patients (111 to 282 months on MHD) and the possible association of genomic damage with the degree of renal insufficiency and duration of MHD. Genomic damage was assessed by evaluating the numbers of micronuclei (MN), which are cytoplasmic DNA-containing structures. The average number of MN in the control group of 23 healthy subjects was 15.3 +/- 4.7 MN/1,000 binucleate (BN) cells. The MN frequencies in the long-term MHD group were significantly greater (44.3 +/- 13.7 MN/1,000 BN) than the control frequencies. A significant increase in MN frequencies (28.2 +/- 9.4 MN/1,000 BN) was also seen in patients with advanced renal failure. The major findings of the study, high MN frequencies in long-term hemodialysis and advanced chronic renal failure patients, may result from decreased DNA repair previously shown and may contribute to the increased cancer incidence in these patients. PMID- 10469853 TI - Comparison of hemostatic factors and serum malondialdehyde as predictive factors for cardiovascular disease in hemodialysis patients. AB - Hemodialysis (HD) patients have accelerated cardiovascular morbidity and mortality rates compared with the general population. Identifying the factors that predict major coronary events in this population can direct the focus on prevention. This cross-sectional study compares known and suspected cardiovascular risk factors in HD patients with and without prevalent cardiovascular disease (CVD). In 76 HD patients (prevalent CVD, 44 of 76 patients), serum lipid, lipoprotein, apolipoprotein (Apo), plasma fibrinogen, tissue plasminogen activator (TPA), plasminogen activator inhibitor (PAI-1), and factor VII levels were measured using standard kits. Serum malondialdehyde (MDA; a marker of oxidative stress) was measured using spectrophotometry. Predictor variables were compared using analysis of variance and chi-squared tests, as appropriate. CVD prevalence was modeled using multiple logistic regression analysis, and odds ratios (OR) were calculated. Serum lipid, lipoprotein, Apo, plasma TPA, PAI-1, and factor VII values did not differ significantly from laboratory norms or discriminate for prevalent CVD in HD patients. Plasma fibrinogen levels were significantly elevated in HD patients compared with laboratory norms (369.4 +/- 130.02 v 276.7 +/- 77.7 mg/dL; P < 0.0001) but were not significantly different in HD patients with and without prevalent CVD. Serum MDA levels, both before and after the midweek HD treatment, were significantly elevated in all HD patients compared with laboratory norms (pretreatment, 2.6 +/- 0.8 nmol/mL; posttreatment, 2.1 +/- 0.3 v 0.91 +/- 0.09 nmol/mL; P < 0.01) and were significantly elevated in HD patients with prevalent CVD versus those without (pretreatment, 2.8 +/- 0.6 v 2.4 +/- 0.4 nmol/mL; P < 0.01; posttreatment, 2.3 +/- 0.4 v 1.94 +/- 0.2 nmol/mL; P < 0.01). Only serum MDA levels, both before and after the midweek treatment, contributed to the explanation of variation in CVD prevalence. OR for CVD in the highest versus lowest tertile of pretreatment MDA level was 2.71 (95% confidence interval [CI], 1.42 to 5.19). ORs for CVD in the highest versus lowest tertile of posttreatment MDA level was 3.65 (95% CI, 1.6 to 8.32). PMID- 10469854 TI - Higher levels of antioxidant defenses in enalapril-treated versus non-enalapril treated hemodialysis patients. AB - We previously reported chronic treatment with angiotensin-converting enzyme inhibitors (ACEis) increases antioxidant defenses in mice. In the present study, however, we examined various antioxidant defenses in chronic hemodialysis (HD) patients either treated with enalapril (10 mg/d) for at least 6 months (+ACEi; n = 11) or untreated (-ACEi; n = 11). The relationship between antioxidant status and HD was investigated by determining oxidative stress markers and antioxidant defenses in a group of chronic HD patients (n = 33) and a group of age-matched controls (n = 29). The effect of a single HD session on those parameters was also evaluated. Before an HD session (pre-HD), HD patients had significantly lower levels of red blood cell (RBC) glutathione (GSH), selenium-dependent glutathione peroxidase activity (RBC-Se-GPx), plasma ubiquinol-10, and alpha-tocopherol than controls. In a randomly selected group of patients (n = 19), a single HD session caused an additional decrease in RBC-GSH and plasma ubiquinol-10 levels. Plasma thiobarbituric acid reactive substance (TBARS) levels were significantly greater in pre-HD patients than controls. Post-HD plasma TBARS levels were similar to control values. The cohort of +ACEi HD patients had greater pre-HD RBC-GSH content, RBC-Se-GPx activity, and plasma beta-carotene concentrations than -ACEi patients (RBC-GSH: +ACEi, 3.1 +/- 0.9 micromol/mL packed RBCs [PRBCs]; -ACEi, 1.2 +/- 0.3 micromol/mL PRBCs [P < 0.05 v +ACEi]; RBC-Se-GPx: +ACEi, 5.8 +/- 0.7 U/mL PRBCs; -ACEi, 4.3 +/- 0.2 U/mL PRBCs [P < 0.05 v +ACEi]; plasma beta-carotene: +ACEi, 0.54 +/- 0.16 micromol/L plasma; -ACEi, 0.19 +/- 0.05 micromol/L plasma [P < 0.05 v +ACEi]). Results show profound alterations in the circulating antioxidant systems of chronic HD patients and that additional oxidative stress occurs during the HD procedure. In addition, in +ACEi HD patients, the levels of several antioxidant defenses are greater than in those in -ACEi HD patients. PMID- 10469855 TI - Effect of calcitriol and age on recovery from hypocalcemia in hemodialysis patients. AB - Calcitriol is used to treat hyperparathyroidism in hemodialysis patients. Calcitriol treatment, either through a reduction in parathyroid hormone (PTH) levels or direct effect on bone, decreases the osteoblast and osteoclast surface and bone formation rate. Our study of 13 hemodialysis patients was designed to evaluate whether calcitriol treatment changed the rate of spontaneous recovery from hypocalcemia induced by a low-calcium dialysis. Calcitriol treatment decreased basal PTH levels from 614 +/- 84 to 327 +/- 102 pg/mL (P < 0.001) and maximal PTH levels from 1,282 +/- 157 to 789 +/- 161 pg/mL (P < 0.001), but the rate of serum ionized calcium recovery from hypocalcemia did not change. When the 13 patients were separated based on the median age of 64 years, the predialysis serum ionized calcium level was less in the younger (group I, 44 +/- 6 years; n = 6) than older (group II, 68 +/- 1 years; n = 7) patients (1.05 +/- 0.03 v 1.22 +/ 0.03 mmol/L, respectively; P < 0.01) despite similar basal (group I, 595 +/- 122 pg/mL v group II, 629 +/- 96 pg/mL) and maximal (group I, 1,114 +/- 299 pg/mL v group II, 1,425 +/- 141 pg/mL) PTH levels. Before calcitriol treatment, the rate of serum ionized calcium recovery from induced hypocalcemia was greater (P < 0.05) for similar PTH levels in the older than younger patients. After calcitriol treatment, despite a similar reduction in PTH levels, the rate of calcium recovery increased (P < 0.05) in the younger patients but did not change in the older patients. We also observed that toward the end of the low-calcium hemodialysis, PTH values decreased even though serum ionized calcium level continued to decline when the rate of calcium reduction slowed. In addition, hysteresis, defined as a lower PTH value during the recovery from hypocalcemia than during the induction of hypocalcemia for the same serum calcium concentration, was present during the spontaneous recovery from hypocalcemia. In conclusion, in the hemodialysis patient: (1) age appeared to affect the bone response to PTH and calcitriol treatment, (2) the PTH response to hypocalcemia was affected by a deceleration in the rate of calcium decrease, and (3) hysteresis of the PTH response to hypocalcemia occurred during the spontaneous recovery from hypocalcemia. PMID- 10469856 TI - Identification of factors responsible for postdialysis fatigue. AB - Ultrafiltration, diffusion, osmotic shifts, blood-membrane interactions, and psychological factors have all been implicated in the pathogenesis of postdialysis fatigue (PDF). To identify responsible factors, we performed a prospective, randomized, crossover analysis of fatigue scores (scale, 0 to 4) in 12 maintenance hemodialysis subjects with PDF. Fatigue scores were evaluated on nondialysis days (baseline) and after the following procedures on midweek treatment days: standard dialysis using either a 135- to 140-mEq/L sodium bath (routine hemodialysis) or a 150- to 155-mEq/L sodium bath (hypernatric hemodialysis); isolated ultrafiltration; isolated diffusion; and sham procedures with (isolated membrane) or without (recirculation) exposure to a dialysis membrane. Maximal fatigue scores are expressed as mean and 95% confidence intervals (CIs). The highest scores were recorded by patients who had just undergone routine hemodialysis (mean score, 2. 6; 95% CI, 1.7 to 3.4), isolated ultrafiltration (mean score, 2.1; 95% CI, 1.3 to 2.9), and isolated diffusion (mean score, 2.4; 95% CI, 1.5 to 3.2). There were no significant differences in fatigue scores between baseline periods and isolated membrane and recirculation procedures (mean score, 1.3; 95% CI, 0.4 to 2.2). Fatigue scores after hypernatric hemodialysis occupied an intermediate position (mean score, 1.7; 95% CI, 0.8 to 2.6). These results suggest that rapid hydraulic and molecular flux have a greater role in the pathogenesis of PDF than psychological stress and blood-membrane interactions. Use of a high-sodium bath may ameliorate PDF. We conclude that appropriate adjustments in both ultrafiltration and sodium profiling remain the most important means for controlling PDF in patients on short-duration hemodialysis. PMID- 10469857 TI - Intradialytic serial vascular access flow measurements. AB - Hemodialysis vascular access failure represents a major source of morbidity and mortality in chronic hemodialysis (CHD) patients. Serial vascular access blood flow (VABF) measurements are being used as a screening method at an increasing rate. There are limited data on the changes in VABF throughout the hemodialysis session, which may potentially affect the validity of VABF measurement. This study is performed to evaluate the trend in VABF during a given hemodialysis session by serial VABF measurements, along with potential factors that may affect VABF. Thirty-two CHD patients had serial VABF measurements performed during a hemodialysis session. Each patient had three serial VABF measurements during a hemodiaysis treatment (within 30, 90, and 150 minutes from the start of hemodialysis). Mean arterial blood pressure (MAP), ultrafiltration rate, and patient symptoms were recorded simultaneously. The mean VABF was 1,344 +/- 486 mL/min within 30 minutes of hemodialysis and decreased to 1,308 +/- 532 and 1,250 +/- 552 mL/min after 90 and 150 minutes, respectively. This trend was statistically significant (P = 0.03). There was a strong correlation between VABF measurements and MAP, which was more pronounced after 90 minutes of initiation of hemodialysis (r = 0.68; P < 0.001). Using multivariate analysis, it can be predicted that after 90 minutes of hemodialysis, each 10% decrease in MAP would result in an expected decrease of 8% in VABF. There was no effect of type of vascular access, baseline VABF, or amount of ultrafiltration on VABF changes. In conclusion, VABF measurements can be performed up to 2 to 2(1/2) hours from the start of hemodialysis in the majority of patients. The major determinant of VABF changes is MAP. In a subset of patients with a decrease MAP greater than 15%, it is advisable to perform VABF measurement either at the first 90 minutes of hemodialysis or postpone it to another treatment session, when MAP is more stable. PMID- 10469858 TI - Does blood flow accurately predict thrombosis or failure of hemodialysis synthetic grafts? A meta-analysis. AB - A number of studies have reported that a single low blood flow (Qa) measurement in synthetic hemodialysis grafts predicts thrombosis or failure. In a meta analysis of these studies, we computed receiver operating characteristic (ROC) curves that evaluated the predictive accuracy of a Qa measurement. The ROC curves plotted sensitivity versus false-positive rate for predicting thrombosis or failure at different Qa thresholds. A perfect predictor has an area under the curve (AUC) of 1.0, whereas a predictor with no discriminative ability has an AUC of 0.5. We identified studies through a literature search and included our own unpublished data. A random-effects model was used to combine the ROC curves from different studies. Of 19 identified studies, 12 were suitable for computing binormal ROC curves (6 predicted thrombosis; 6 predicted failure). The studies measured Qa and then observed outcome during periods of 1.5 to more than 6 months. The combined AUCs from these studies indicate Qa was a relatively poor predictor, with 0.70 +/- 0. 04 (range, 0.61 to 0.84) for thrombosis and 0.76 +/- 0.07 (range, 0. 62 to 0.90) for failure. The wide range of AUCs also shows there was much heterogeneity between studies. We conclude that a single Qa measurement does not appear to have enough accuracy to be a clinically useful predictor of graft thrombosis or failure. Serial Qa measurements and identification of factors that caused heterogeneity between studies may be needed to achieve sufficient accuracy. PMID- 10469859 TI - Diffusive and convective solute clearances during continuous renal replacement therapy at various dialysate and ultrafiltration flow rates. AB - Clearances of several solutes (urea, creatinine, phosphate, urates, beta(2) microglobulin [beta(2)-M]) were measured during venovenous continuous renal replacement therapy (CRRT) at various ultrafiltration (Q(UF); 0 to 2 L/h) and dialysate flow rates (Q(D); 0 to 2.5 L/h). Preset Multiflow-60 and Multiflow-100 hollow-fiber dialysers (M-60 and M-100; Hospal-Gambro, St-Leonard, Canada) were compared (five patients for each type). First, we evaluated the impact of predilution on convective clearances: a progressive decrease in patient clearances, similar for both filters, was observed, reaching a maximum of 15%, 18%, and 19% for urea, urates, and creatinine, respectively, with predilution at a Q(UF) of 2 L/h. Second, we compared convective and diffusive clearances. Because effluent to plasma ratio (E/P) remained at 1 for small solutes (urea, creatinine, phosphate, urates) during convection, clearances were equal to the effluent rate for both dialyzers. However, we observed greater diffusive clearances for small molecules with M-100 than with M-60 at a Q(D) of 1.5 to 2.5 L/h, the difference being more significant as molecular weight increased. For beta(2)-M, diffusive clearance was very low and rapidly reached a plateau of 8 and 12 mL/min for M-60 and M-100, respectively, at a Q(D) greater than 1.5 L/h. Convective clearances for beta(2)-M increased nonlinearly up to 20 +/- 2 mL/min at a progressively greater Q(UF) (from 0.5 to 2 L/h) for both M-60 and M-100. This nonlinear increase was attributed to an increase of almost 40% in E/P for beta(2)-M from a Q(UF) of 0.5 to 2 L/h. Third, the interaction between convection and diffusion was assessed by measuring solute clearances at a fixed Q(UF) (1 and 2 L/h) and variable Q(D) (0.5 to 2.5 L/h). For small molecules, no significant interaction between convection and diffusion was noticed with M-100, whereas only a small interaction was noticed with M-60. However, for beta(2)-M, the addition of diffusion (Q(D), 0.5 to 2.5 L/h) did not result in any significant increase in total clearances over convective clearances for M-60 and M-100. This observation suggests that the diffusive clearances for beta(2)-M observed with M-60 and M-100 at a Q(UF) of 0 L/h and at various Q(D) probably occurs by convective fluxes across the membrane. These results demonstrate that convection is more efficient than diffusion in removing mixed-molecular-weight solutes during CRRT. PMID- 10469860 TI - Association of acidosis and nutritional parameters in hemodialysis patients. AB - There is extensive literature supporting an important role for acidosis in inducing net protein breakdown, both in experimental animals and humans. However, the clinical importance of the moderate intermittent metabolic acidosis frequently observed in hemodialysis patients has not been determined. We performed a cross-sectional analysis of the baseline laboratory data in the first 1,000 patients recruited to the Hemodialysis Study, looking for correlations between predialysis serum total carbon dioxide levels and parameters related to dietary intake and nutritional status. We found the mean predialysis serum total carbon dioxide level was moderately low (21.6 +/- 3.4 mmol/L; mean +/- SD) despite the use of bicarbonate dialysate and an average single-pool Kt/V of 1.54. Predialysis serum total carbon dioxide level correlated negatively with normalized protein catabolic rate (P < 0.001), suggesting patients with lower serum total carbon dioxide levels have a greater protein intake. The degree of acidosis observed in our patients does not seem to have a deleterious effect on the nutritional status of these patients because correlation of serum total carbon dioxide level with nutritional parameters, such as serum creatinine and serum albumin levels, was either negative or not statistically significant. Further investigation of the effect of modifying serum bicarbonate concentration on nutritional markers is needed to test these hypotheses. PMID- 10469861 TI - Outcome of nutritional status and body composition of uremic patients on a very low protein diet. AB - Concern has been raised about the nutritional adequacy of a very low protein diet (VLPD). Monthly clinical evaluation by a physician and dietitian and quarterly dietary records, anthropometric measurements, blood testing, and dual energy X ray absorptiometry (DEXA) were used to assess the course of nutritional status for 1 year in 10 clinically stable patients (six men, four women; age, 57.1 +/- 9.3 years) with advanced chronic renal failure (mean glomerular filtration rate, 13.2 +/- 4.8 mL/min/1.73 m(2)). These patients received a VLPD providing 0.3 g/kg/d of protein and were supplemented with amino acids and ketoanalogues. Conventional nutritional markers remained unchanged after 1 year of the VLPD. However, during the same period, whole-body DEXA showed a significant decrease in lean tissue from 46.2 +/- 10.2 to 45.0 +/- 9. 8 kg (P < 0.02); limb-trunk lean tissue ratio was reduced from 0.86 +/- 0.12 to 0.82 +/- 0.12 (P < 0.02), total body fat increased from 20.0 +/- 6.9 to 21.4 +/- 7.0 kg (P < 0.05), and the percentage of total-body fat increased from 29.2% +/- 8.7% to 31.7% +/- 8.8% (P < 0.03). These different modifications occurred abruptly during the first 3 months, then stabilized or slightly improved thereafter. These mild changes do not appear to be deleterious given the favorable long-term outcome of these patients, even after they began treatment by dialysis or after renal transplantation. PMID- 10469862 TI - Diagnosis of iron deficiency in chronic renal failure. AB - The cause of anemia in chronic renal failure is multifactorial. Decreased erythropoietin (EPO) production is the main pathogenetic factor, but iron deficiency is the primary cause of unresponsiveness to EPO therapy. The diagnosis of iron deficiency in patients with chronic renal failure is difficult. We assessed the sensitivity and specificity of serum ferritin, total iron-binding capacity, transferrin saturation index, erythrocyte ferritin, and serum transferrin receptor in 63 patients with chronic renal failure undergoing dialysis (47 men, 16 women) with iron deficiency anemia. They were selected on the basis of clinical stability and absence of factors that may interfere with iron metabolism. None of the patients had received intravenous iron therapy or recombinant human erythropoietin (rHuEPO). Bone marrow biopsy with iron staining was the reference standard for iron stores. The receiver operating characteristic (ROC) curve and the area under the curve were calculated to assess the sensitivity and specificity of iron metabolism parameters. The parameter with the largest area under the ROC curve was serum ferritin (0.83). A cut point of 121 microgram/L showed a sensitivity and a specificity of 75%. The areas under the ROC curves of serum transferrin receptor and erythrocyte ferritin were 0.69 and 0.68, respectively. The remaining parameters showed areas under the ROC curve less than 0.65. Although serum transferrin receptor and erythrocyte ferritin may be acceptable markers for iron deficiency in stable chronic renal failure patients, serum ferritin level continues to be the most reliable diagnostic parameter. Transferrin saturation index is not a reliable parameter for the diagnosis of iron deficiency in stable patients not treated with rHuEPO. PMID- 10469863 TI - Clinical and biochemical characteristics of type 2 diabetic patients on continuous ambulatory peritoneal dialysis: relationships with insulin requirement. AB - Although glycemic control has an important impact on the clinical outcomes of patients with diabetes undergoing dialysis, there is a paucity of data on the relationship between glucose metabolism and clinical parameters in these patients. In this study, we compared a cohort of 48 patients with type II diabetes undergoing continuous ambulatory peritoneal dialysis (CAPD) with 84 age- and sex-matched patients with type II diabetes with similar disease duration but normal renal function. Compared with those with normal renal function, patients with type 2 diabetes undergoing CAPD had greater serum angiotensin-converting enzyme activity (median, 57.4 U/L; range, 33.5 to 100.0 U/L v 46.9 U/L; range, 11.6 to 111.2 U/L; P < 0.005), fasting C-peptide (median, 9.1 ng/mL; range, 0.9 to 30.0 ng/mL v 2.2 ng/mL; range, 0.2 to 20.3 ng/mL; P < 0.0001) and triglyceride levels, and lower serum albumin concentrations. Among the patients undergoing CAPD, there was a preponderance of men in the insulin-treated group. Insulin treated patients also had greater plasma albumin levels and body weights and lower fasting serum C-peptide levels (2.81 +/- 1.77 v 3.12 +/- 2.04 ng/mL; analysis of variance, P = 0.007 adjusted for fasting glucose concentration). Multivariate analysis showed duration of diabetes, hemoglobin A(1c) (HbA(1c)) level, and body weight were independent determinants of insulin requirement in patients undergoing CAPD. The daily insulin dosage required was related to the duration of diabetes (r = 0.5; P = 0.007). In summary, among patients with end stage renal failure, insulin-treated patients had greater body weights and plasma albumin levels but lower cholesterol levels. Plasma C-peptide concentration and duration of diabetes were the main determinants of insulin requirement, reflecting a decrease in beta-cell reserve, whereas the daily insulin dose correlated mainly with body weight, HbA(1c) level, and duration of diabetes. Kt/V had no effect on insulin resistance or insulin requirement of the patients. PMID- 10469864 TI - Does immunosuppression with prednisolone and azathioprine alter the progression of idiopathic membranous nephropathy? AB - The role of immunosuppressive drugs in the treatment of idiopathic membranous nephropathy (IMN) remains controversial. The effect of treatment with prednisolone and azathioprine in patients with nephrotic-range proteinuria and biopsy-proven IMN from a single center (Sheffield Kidney Institute, Sheffield, UK) is described. In this retrospective study, 58 patients with IMN and nephrotic range proteinuria were followed up for 4 years. Thirty-eight patients were treated with prednisolone (1 mg/kg body weight/d) and azathioprine (2 mg/kg body weight/d) orally for a median period of 26 months (range, 6 to 48 months). Twenty patients received no specific treatment for IMN and served as a control group. Clinical, biochemical, and histopathologic features at presentation were similar between the groups. Renal function (RF), measured by serum creatinine (Scr) level, deteriorated in both treated and control groups during the follow-up period. The median initial and final Scr levels (at the end of follow-up) in the treated group were 1.6 and 2. 1 mg/dL, respectively, and in the control group were 1.3 and 1.7 m/dL, respectively (P = not significant). Neither the rate of RF decline (measured by the slope of reciprocal of Scr against time) nor the proportion of patients with deteriorating RF differed significantly between the groups (37%, treated group; 30%, control group). A significant reduction in proteinuria was observed in both groups (P < 0.01, either group). Also, the rate of remission of nephrotic-range proteinuria was not significantly different between groups (55%, treated group; 65%, control group). The only prognostic factor that correlated with RF outcome (expressed by final Scr level) in a given patient was the mean proteinuria during follow-up in either group (r = 0.493; P < 0.01, treated group; r = 0.651; P < 0.01, control group). Adverse effects of immunosuppressive treatment were observed in nine patients (24%). These were serious in four patients (10%) and included squamous cell carcinoma (two patients), bacterial meningitis (one patient), and septicemia (one patient). In conclusion, treatment with prednisolone and azathioprine for patients with IMN did not show significant beneficial effects on the progression of disease. Furthermore, this treatment was associated with frequent and serious adverse effects. PMID- 10469865 TI - Clinical and prognostic value of serial renal biopsies in lupus nephritis. AB - Little information is available about the role of repeated renal biopsies in lupus nephritis. We analyzed retrospectively the prognostic significance of serial renal biopsies in patients with lupus nephritis. Thirty-one patients with lupus nephritis underwent two or more renal biopsies during follow-up. The indications for repeated biopsy were as follows: improvement of renal disease but persistence of nonnephrotic proteinuria (group A, 7 patients); persistent or relapsing nephrotic syndrome (group B, 12 patients); and worsening of renal function (group C, 19 patients). After a median follow-up of 10.5 years, 17 patients reached the end point (persistent doubling of plasma creatinine level). At repeated renal biopsy, there was a correlation between improved clinical and histological features for group A. In these patients, treatment was reduced or stopped successfully. Histological features remained almost unchanged in group B. All patients showed an improvement of proteinuria after reinforcement of therapy. In group C, the worsening of renal function was associated with a variable and clinically unpredictable combination of active and chronic lesions. Only the few patients with an elevated activity index and moderate chronicity index showed a favorable and persistent improvement of renal disease after reinforcement of therapy. At multivariate analysis of clinical and histological data at presentation, only male sex was predictive of an adverse outcome (P = 0.015). At repeated renal biopsy, crescents in more than 30% of glomeruli (P = 0.0009) and chronicity index of 5 or greater (P = 0.00006) were associated with the probability of reaching the end point at multivariate analysis. Repeated renal biopsy may be helpful for establishing the prognosis in patients with lupus nephritis, particularly in the presence of worsening of renal function. PMID- 10469866 TI - Drug-induced acute interstitial nephritis in renal allografts: histopathologic features and clinical course in six patients. AB - Drug-induced acute interstitial nephritis is a common cause of dysfunction in native kidneys, but is rarely reported in renal allografts. This report describes six renal transplant recipients with acute renal allograft dysfunction or delayed allograft function in whom a renal transplant biopsy showed histopathologic features of drug-induced interstitial nephritis with no diagnostic evidence of acute rejection, cyclosporine or tacrolimus nephrotoxicity, or other lesion that could account for the graft dysfunction. In five of the six patients, interstitial nephritis occurred within 4 weeks of transplantation. All the patients were receiving trimethaprim-sulfamethoxazole and/or other drugs associated with interstitial nephritis. After discontinuation of these drugs and short-term corticosteroid treatment, all patients showed improvement in renal function, although the time course of this improvement varied considerably, with three patients showing a return to baseline serum creatinine level within 2 weeks and two patients showing a gradual improvement over 8 weeks. Four of the five patients followed up for more than 1 year (range, 14 to 33 months) after the episode of interstitial nephritis had good allograft function (serum creatinine level NNP > Control) for several measures of gait variability, most of these trends were not statistically significant. We hypothesize that motorized treadmill walking may be inherently less variable than overground walking and that statistical measures of variability may not be sufficient to fully characterize stride-to-stride variability in human locomotion. PMID- 10469939 TI - The coordinated movement of the lumbo-pelvic-hip complex during running: a literature review. AB - The purpose of this review article is to summarise the literature to date regarding the movement of the lumbar spine, pelvis and hips during running gait. Both two-dimensional and three-dimensional studies are analysed to illustrate the apparent coordination in the angular kinematics of each of these segments during running. Knowledge of this coordination is essential in order to facilitate the successful rehabilitation of running injuries to the back, pelvis, hip and thigh. PMID- 10469940 TI - [Factors related to treatment adherence in schizophrenic patients]. AB - BACKGROUND: The percentage of lack of adherence in schizophrenia is estimated in 50%. Non-compliance cause higher rates of relapses, rehospitalizations, suicide, homicide and violent behaviour among schizophrenic patients. The aim of this study was to find several factors related to adherence in schizophrenia. MATERIAL AND METHODS: 40 schizophrenic patients were assessed with a protocol which examined sociodemographic, clinical and therapeutical variables, scales in order to evaluate insight (Birchwood), drug attitude (Drug Attitude Inventory), psychopathology (BPRS) and side effects (UKU). Variables of outcome and overall functioning (GAS) in the previous year were also included. RESULTS: Multiple regression analysis revealed that BPRS thought disorder subscale and UKU neurologic side effects subscale predicted 24% of the variance of adherence. Adherence is related to insight and attitude to medication too. An adequate adherence was related to better overall functioning, less psychopathology and fewer hospitalizations during the year prior to the study. PMID- 10469941 TI - [A study on suicide in Asturias: increase of the frequency in the last two decades]. AB - INTRODUCTION: The suicide is actually a significant public health issue. MATERIALS AND METHODS: The present work studies the mortality rates by suicide registered in Asturias between the years 1975 and 1994, being based on the data published by National Statistics Institute. RESULTS: The average annual suicide rate of the period was of 10.75 per 100,000 inhabitants, with a proportion man/woman of 2.3/1. The suicide rate increases with the age, so that the suicides of people older than 70 years meant 27% of the total of suicides in man and 36% of the total in women. A gradual increase of the phenomenon was evidenced in the temporary analysis, the standardized average annual rate went from 7.51 in 1974 to 12.91 in 1994. The annual rates increased in all the groups of age, but especially in young men and women in the decade of 40. The most used method of suicide was hanging, followed by throwing and immersion. It was evidenced a temporary trend to the increase of suicides by poisoning, shot by gun and throwing from a high place. DISCUSSION AND CONCLUSIONS: The raise of suicide rates could be influenced by sociological factors, desinstitutionalization in mental health and accessibility to suicide methods. It is important to design prevention programs adapted to local characteristics. PMID- 10469942 TI - [Subsyndromal depressive semiology in severe alcoholism]. AB - INTRODUCTION: Several investigations have communicated frequent association between alcohol dependence and depression. METHOD: 21 subjects with DSM-IV criteria for alcohol dependence were included in an open label trial for alcohol withdrawal. At inclusion and along the follow-up none of the probands met DSM-IV criteria for mood disorder. Follow-up included a 15-day detoxification period and 195 days of withdrawal program, including treatment with 20 mg/d of fluoxetine. Occurrence of depressive semiology was measured using the Beck Depression Inventory (BDI) on day 15 (after detoxification), day 75 and day 210 (after withdrawal from ethanol of 195 days). RESULTS: 67% of the sample showed a positive basal BDI (after-detoxification-BDI> 9). Global retention rate after 210 days of follow-up was 57%. All patients who dropped out the investigation before completing the protocol showed a basal BDI in the depressive rank (BDI= 10-63), and maintained depressive scores in this instrument until their abandonment. CONCLUSIONS: Prevalence of <> in this population appears to be high. This clinical feature is frequently ignored because most of the patients do not meet standardized diagnostic criteria for mood disorders. Post-detoxification BDI could be used as a predictive factor of therapeutic result in long-term alcohol withdrawal programs. In addition, in our study fluoxetine showed efficacy in maintaining long-term alcohol abstinence. PMID- 10469943 TI - [Quality of life and disability in chronic schizophrenics treated with with risperidone and previously treated with depot neuroleptics]. AB - OBJECTIVES: 1. Assess the evolution of disability and quality of life in schizophrenic patients treated with risperidone and who had received depot neuroleptics; 2. Evaluate risperidone efficiency; 3. Evaluate safety of this drug. METHOD: Post-marketing multicentric observational (8 months) surveillance study was carried out. PATIENTS: 109 schizophrenic patients (ICD-10 criteria). ASSESSMENTS: baseline and months 2, 4 and 8. INSTRUMENTS: SF-36, WHO/DDS-S, BPRS and CGI. Safety was evaluated by the UKU subscale for neurological side effects and spontaneous reports. RESULTS: statistically significant improvement of the mean scores of BPRS, CGI, WHO/DDS-S and SF-36 at 2, 4 and 8 months. There was a significant reduction in the total UKU subscale for neurological side effects scores from visit 1 (month 2) onwards. Risperidone was generally well tolerated by the study patients. From a total of 104 patients included, only 4 (3.8%) discontinued treatment due to adverse reactions. During the 8 months of study period, 87.6% of the patients did not suffer any adverse event; the resting 12.4 suffered one or more side effects. The most frequently reported adverse events according spontaneous reports were: anxiety and restlessness (n= 4; 3.8%), weight increase (n= 4; 3.8%), sexual disturbances (n= 4; 3.8%) and amenorrhea (n= 2; 1.9%) among others. CONCLUSION: the long-term treatment with risperidone has improved the disability and quality of life levels of a large group of schizophrenic patients previously treated with depot neuroleptics. PMID- 10469944 TI - [Alcohol dependent subjects with psychiatric comorbidity: a heterogeneous group]. AB - METHODOLOGY: In a sample of 161 alcohol dependent outpatients, we carry out three comparative analysis between alcohol dependent subjects with and without associated affective disorders (n= 72); with and without associated personality disorders (n= 60) and with and without associated substance abuse disorders (n= 26) based on sociodemographic, clinical characteristics, treatment response and long-term evolution variables. FINDINGS: As for associated affective disorders, the only difference is that there are more female. As for associated personality disorders, we find that they are younger, there are more female, they have previous treatment background and worse short-term response. As for the group with associated substance abuse disorders, they are younger, have lower educational and social economic level, alcohol dependence onset earlier, more legal complications and worse half-term treatment response. CONCLUSIONS: The alcohol dependent patients with comorbidity are a heterogeneous group which makes the use of it difficult to categorize alcohol dependence. PMID- 10469945 TI - [Styles and types of quality of life in the Seville Questionnaire]. AB - The Sevilla Scale of Quality of Life is composed by two scales: the Scale F or scale of favourable aspects and the Scale D or scale of unfavourable aspects. This work elaborates some conceptual procedures and some operative strategies to interpret in a complex way the quality of life. We build a unique and global punctuation for the CSCV that we denominate PCV, and that it is the result of other partial scores; we also outline the analysis of the dispersion matrix of the PCV like a system to differentiate styles different from quality of life. We use the factor structure of the CSCV to represent each subject in a three dimensional space by means of two points, the favourable aspect and the unfavourable one; the geometric distance between both points represent another conceptual shade to interpret of the types of quality of life. The necessity is underlined of guaranteeing the utility of these procedures by means of future empiric analysis, with samples of subject so much normal as pathological. PMID- 10469946 TI - [Causes of failure in psychopharmacological treatment of anxiety disorder]. AB - OBJECTIVE AND METHODS: A comprehensive review of literature on the causes of failure in pharmacologic treatment of panic disorder (PD). RESULTS: PD has high rates of response to psychopharmacological treatment, reaching up to 80-90%. However, in a small percentage of cases (10-20%), the disorder is refractory to the combination of drugs and psychotherapy, at correct dosages and during adequate time periods. Refractoriness in PD is not a well-known issue as few studies have addressed this question. Intolerance to psychotropic drugs is usually reported to be the commonest cause, especially to antidepressants. Other causes are: comorbidity (especially with depressive disorders, personality disorders, and alcohol abuse, as well as with medical conditions); severe panic attacks; severe agoraphobia; and long illness duration. <> PD is to be ruled out, since incorrect diagnosis, incorrect treatment, or bad compliance are common causes of a poor response. We review treatment approaches for PD when no response is achieved with antidepressants (TCA, SSRI, MAOI), alprazolam and cognitive-behaviour therapy. Two recommended strategies are drug potentiation and use of clonazepam. PMID- 10469947 TI - [Applications of the magnetoencephalography in the study of schizophrenia]. AB - In this paper we present a revision about magnetoencephalography (MEG) and schizophrenia. We focus on Auditive Magnetic Fields, principally in M100 and in his hemispheric asymmetries. Finally, it was discussed the advantages of the MEG in comparison with other techniques with low temporal (RMf, PET) or spatial (EEG) resolution. PMID- 10469948 TI - [Neurodevelopmental hypothesis in functional psychosis]. AB - Evidence from several studies supports the neurodevelopmental hypothesis of schizophrenia. Others suggest a substantial body of evidence attesting to a possible neurodevelopmental origin of affective disorders. However, there is little evidence that any risk factor or neurodevelopmental marker is specific to any diagnostic category within functional psychoses. Some authors have proposed a new model which implies a continuity within the category of psychosis based on a psychopathological continuum. This research strategy has important and useful advantages which are discussed in this review. PMID- 10469949 TI - [Clomipramine in the treatment of catatonia]. AB - The use of intravenous clomipramine gave rise to the remission of catatonia on the third or fourth day of treatment in two patients diagnosed as suffering from affective disorders. Therapeutic effect was maintained on switching to oral clomipramine. Clomipramine could be a therapeutic alternative in the management of catatonia associated with affective disorders. PMID- 10469950 TI - Honored Guest's Address. A practical affair. PMID- 10469951 TI - Time-dependent cellular population of textured-surface left ventricular assist devices contributes to the development of a biphasic systemic procoagulant response. AB - OBJECTIVE: Textured-surface left ventricular assist devices (LVAD) have been shown to enhance ventricular function and survival in patients with end-stage heart failure. Furthermore, we have described a procoagulant physiology in our LVAD population with sustained thrombin generation (elevated thrombin antithrombin III complex and prothrombin fragment 1+2) and fibrinolysis (D dimers), even up to 335 days after LVAD placement. To explain such sustained activation of coagulation, we speculated that the LVAD surface selectively adsorbed and promoted activation of circulating blood cells. METHODS: In a prospective study of 20 patients with LVADs, we examined samples of peripheral blood as well as cells harvested from the surface of the LVADs at the time of their explantation for procoagulant proinflammatory markers. RESULTS: Analysis of the cells populating the LVAD surface revealed the presence of pluripotent hematopoietic CD34(+) cells, as well as cells bearing monocyte (CD14)/macrophage (CD68) markers, which also expressed procoagulant tissue factor. Reverse transcriptase-polymerase chain reaction confirmed cellular activation on the LVAD surface, revealing transcripts for interleukin 1alpha, interleukin 2, and tumor necrosis factor alpha, in addition to vascular cell adhesion molecule-1 consistent with their capacity to continually recruit and activate circulating cells, thereby propagating their response. In the periphery, elevated levels of tissue factor were found in the plasma of patients with LVADs, along with enhanced procoagulant activity. CONCLUSION: These observations suggest that the LVAD surface selectively absorbs and activates circulating hematopoietic precursor and monocytic cells, thereby creating a sustained prothrombotic and potentially proinflammatory systemic environment. PMID- 10469953 TI - Commentary PMID- 10469952 TI - Is cardiopulmonary bypass still the cause of cognitive dysfunction after cardiac operations? AB - OBJECTIVE: The purpose of this study was to determine whether cognitive impairment is related to cardiopulmonary bypass. METHODS: Twenty-five patients undergoing coronary artery bypass grafting without cardiopulmonary bypass were matched with 50 patients undergoing coronary artery bypass grafting with cardiopulmonary bypass. All patients received the same anesthetic regimen, and one surgeon performed all the operations. A battery of 10 standard tests of neuropsychologic function were performed before, at discharge, and 3 months after the operation. A comprehensive multidimensional measure of subjective health status was used as the primary clinical measure of functional outcome. RESULTS: The groups were similar with respect to age, sex, and ventricular function and differed only in the need for a circumflex artery graft. Both groups showed significant improvement in the comprehensive multidimensional measure of subjective health status at 3 months. At discharge most neuropsychologic tests had deteriorated in both groups (the same 4 tests had deteriorated significantly in both groups, and an additional test had deteriorated significantly in the cardiopulmonary bypass group). At 3 months all but one test in the cardiopulmonary bypass group had returned to or exceeded baseline performance. The same 2 tests had improved significantly in both groups, and a further test had improved significantly in the group without cardiopulmonary bypass. At no specific time point was there a significant difference between the absolute or change scores between the groups on any of the tests. CONCLUSIONS: The similar pattern of early decline and late recovery of cognitive function in patients undergoing coronary artery bypass grafting with and without cardiopulmonary bypass suggests that cardiopulmonary bypass is not the major cause of postoperative cognitive impairment. This merits consideration in deciding optimal treatment strategies in coronary revascularization. PMID- 10469955 TI - Commentary PMID- 10469954 TI - Cardiopulmonary bypass and activation of antithrombotic plasma protein C. AB - OBJECTIVE: We hypothesized that antithrombotic plasma-activated protein C plays a defensive antithrombotic role during coronary ischemia and postischemic reperfusion. METHODS AND RESULTS: We evaluated protein C activation during cardiopulmonary bypass and coronary reperfusion in 20 patients undergoing coronary bypass surgery. During cardiopulmonary bypass and during the 10 minutes after aortic unclamping, the plasma levels of protein C (mean +/- standard error of the mean) decreased from 123% +/- 7% to 74% +/- 5% of normal mean. In contrast, the levels of activated protein C in plasma increased from 122% +/- 8% to 159% +/- 21%, and the activated protein C/protein C ratio increased from 1.04 +/- 0.08 to 2.29 +/- 0. 31 (P =.006, 2-tailed Wilcoxon signed rank test). Patients were stratified on the basis of the increase in activated protein C in the coronary sinus plasma at 10 minutes after reperfusion by means of the arbitrary value of 1.5 for the activated protein C/protein C ratio. Within 24 hours, the patients with low increases in activated protein C (ratio < 1.5, n = 8) had a significantly (P <.05) lower cardiac output and mean pulmonary artery pressure, as well as a higher systemic vascular resistance, than patients (n = 11) with high increases in activated protein C (ratio > 1.5). The rapid increase in activated protein C during the first 10 minutes after aortic unclamping indicated protein C activation in the reperfused vascular beds. CONCLUSIONS: The antithrombotic protein C pathway was significantly activated during cardiopulmonary bypass mainly during the minutes after aortic unclamping in the ischemic vascular beds. Suboptimal protein C activation during ischemia may impair the postischemic recovery of human heart and circulation. PMID- 10469956 TI - Superior extension of intraoperative brain damage in case of normothermic systemic perfusion during coronary artery bypass operations. AB - OBJECTIVE: Despite the controversies on the potential detrimental effects of normothermic cardiopulmonary bypass on neurologic outcome, to date no correlation between the severity of intraoperative brain lesions and the cardiopulmonary bypass temperature used at operation has been reported. This study compares the prevalence and the severity of brain lesions in patients who underwent operation in condition of normothermic versus hypothermic systemic perfusion. METHODS: Data are derived from the analysis of 2987 consecutive primary isolated myocardial revascularizations performed at our institution between April 1990 and January 1997. Of these cases, 1385 procedures were hypothermic and 1602 procedures were normothermic systemic perfusion. In all cases the neurologic outcome and extent of ischemic areas were prospectively recorded. RESULTS: Overall, 31 patients had a perioperative stroke (1.0%). The prevalence of neurologic events was similar in the 2 groups (15 cases in the hypothermic group and 16 cases in the normothermic perfusion group; P, not significant). However, the mean Glasgow Outcome Scale score and computed tomography-demonstrated extent of brain lesions were significantly worse in the normothermic group. CONCLUSIONS: Although the prevalence of intraoperative stroke was similar with hypothermic or normothermic cardiopulmonary bypass, the use of normothermic systemic perfusion was associated with more extended brain damage at computed tomographic scan and with a worse neurologic outcome. These results demand caution in the use of normothermic cardiopulmonary bypass and claim further investigation on the neurologic safety of normothermia. PMID- 10469957 TI - Preconditioning improves cardioplegia-related coronary microvascular smooth muscle hypercontractility: role of KATP channels. AB - OBJECTIVES: The effect of preconditioning before hyperkalemic cardioplegia on the coronary smooth muscle remains to be elucidated. We tested the hypothesis that hypoxic preconditioning could protect coronary smooth muscle against subsequent hyperkalemic cardioplegia-induced coronary vasospasm and that this preconditioning effect could be mediated by K(ATP) channels. METHODS: Rat coronary arterioles (endothelium-denuded) were studied in a pressurized, no-flow, normothermic state. Simultaneous monitoring of luminal diameter and intracellular calcium concentration of vascular smooth muscle loaded with fura-2 was made with microscopic image analysis. All vessels were subjected to 60 minutes of hypoxic hyperkalemic cardioplegia (K(+) = 25.0 mmol/L) and were then reperfused. Six groups were studied: (1) controls, no precardioplegic intervention; (2) preconditioning, achieved with 10 minutes of hypoxia (PO2 < 30 mm Hg) and 10 minutes of reoxygenation; (3) preconditioning plus glibenclamide (10 micromol/L), achieved with 10 minutes of preconditioning in the presence of K(ATP) channel blocker glibenclamide; (4) pretreatment with K(ATP) channel opener pinacidil (100 micromol/L); (5) pretreatment with pinacidil (100 micromol/L) plus glibenclamide (10 micromol/L); and (6) pretreatment with glibenclamide (10 micromol/L) alone. RESULTS: Hypoxic preconditioning significantly (P <.01) reduced hyperkalemic cardioplegia-induced intracellular calcium concentration accumulation and prevented the hypercontractility during and after hyperkalemic cardioplegia compared with control vessels. Pinacidil provided effective microvascular protection similar to hypoxic preconditioning. These vasoprotective effects of preconditioning were significantly antagonized in glibenclamide-treated vessels. CONCLUSIONS: Hypoxic preconditioning can prevent coronary microvascular hypercontractility during and after subsequent cardioplegia by a K(ATP ) channel mechanism that regulates intracellular calcium concentration of the vascular smooth muscle. PMID- 10469958 TI - Gene transfection of beta 2-adrenergic receptor into the normal rat heart enhances cardiac response to beta-adrenergic agonist. AB - BACKGROUND: Beta-adrenergic receptor system has a major role in cardiac contraction. If the receptor can be increased by gene transfection by means of intracoronary infusion of beta 2-adrenergic receptor to the hearts in which the receptor is down-regulated, this maneuver may improve the cardiac function and may be applied as one therapeutic approach during cardiopulmonary bypass or percutaneous cardiopulmonary support. METHODS AND RESULTS: The beta 2-adrenergic receptor complementary DNA was transfected in vivo to the normal rat heart by intracoronary infusion by means of a hemagglutinating virus of Japan liposome method, and the transfected heart was transplanted into the abdomen of another rat. Four days after transfection, the sarcolemma of the cardiomyocytes was well labeled by immunohistochemical labeling. Expression of beta-adrenergic receptor in the heart was approximately 4 times greater than that in control hearts (134 +/- 42 vs 33 +/- 4 fmol/mg protein) according to a ligand binding assay. The cardiac response of the transfected heart to isoproterenol was shown to be enhanced in a Langendorff perfusion system: after isoproterenol, developed pressure and maximal derivative of the left ventricle were greater than in the control heart (200 +/- 12 vs 174 +/- 6 mm Hg and 4110 +/- 130 vs 3491 +/- 255 mm Hg/sec), and the minimal derivative of the left ventricle was markedly smaller ( 3040 +/- 267 vs -2528 +/- 131 mm Hg/sec). CONCLUSIONS: These results indicate that expression of beta 2-adrenergic receptor was approximately 4 times greater than in normal rat hearts by gene transfection using a hemagglutinating virus of Japan liposome method, and the transfected hearts demonstrated marked enhancements in cardiac response to beta-agonist, suggesting that transfer of this gene by intracoronary infusion has potential as a novel approach to enhance cardiac function. PMID- 10469959 TI - Release of cardiac troponin I in antegrade crystalloid versus cold blood cardioplegia. AB - OBJECTIVE: The purpose of this study was to assess the efficacy of myocardial protection, comparing antegrade crystalloid cardioplegia with cold blood cardioplegia, in patients with preserved left ventricular function who were undergoing elective first coronary artery bypass grafting. Release of cardiac troponin I was used as a marker for the effectiveness of myocardial protection. METHODS: A consecutive series of 62 patients were randomly assigned to receive crystalloid or blood cardioplegia. Cardiac troponin I concentrations were determined in venous blood samples before the operation, immediately after unclamping, at 6, 9, 12, and 24 hours, and daily thereafter for 5 days. RESULTS: Rising levels of troponin I were found in all patients. The time course and peak release were similar in the crystalloid cardioplegia and the blood cardioplegia groups. No patients in either group had electrocardiographic evidence of perioperative myocardial infarction. Cardiac troponin I was able to detect small areas of myocardial damage, not revealed by electrocardiography or creatine kinase MB release. Aprotinin administration was associated with lower cardiac troponin I release in both groups. Cardiac troponin I was lower in patients whose conditions did not require electrical defibrillation after aortic unclamping, irrespective of cardioplegia type. The presence of a main stem lesion was associated with higher cardiac troponin I release only in the crystalloid cardioplegia group. CONCLUSIONS: Antegrade cold blood cardioplegia is equally effective as antegrade crystalloid cardioplegia in a randomized group of patients with preserved left ventricular function who were undergoing elective first coronary artery bypass grafting. Aprotinin administration resulted in lower cardiac troponin I release, whereas electrical defibrillation was related to a higher release irrespective of cardioplegia type. The presence of a main stem lesion resulted in higher cardiac troponin I release in the crystalloid cardioplegia group. PMID- 10469960 TI - Selective blockade of membrane attack complex formation during simulated extracorporeal circulation inhibits platelet but not leukocyte activation. AB - OBJECTIVE: Complement activation is induced by cardiopulmonary bypass, and previous work found that late complement components (C5a, C5b-9) contribute to neutrophil and platelet activation during bypass. In the present study, we blocked C5b-9 formation during extracorporeal recirculation of whole blood to assess whether the membrane attack complex was responsible for both platelet and leukocyte activation. METHODS: In a simulated extracorporeal model that activates complement (C3a and sC5b-9), platelets (CD62P expression, leukocyte-platelet conjugate formation), and leukocytes (increased CD11b expression and neutrophil elastase), we examined an anti-human C8 monoclonal antibody that inhibits C5b-9 generation for its effects on cellular activation. RESULTS: Anti-C8 significantly inhibited sC5b-9 formation but did not block C3a generation. Anti-C8 also significantly inhibited the increase in platelet CD62P and monocyte-platelet conjugate formation seen with control circulation. Moreover, compared with control circulation, in which the number of circulating platelets fell by 45%, addition of anti-C8 completely preserved platelet counts. In contrast to blockade of both C5a and sC5b-9 during simulated extracorporeal circulation, neutrophil activation was not inhibited by anti-C8. However, circulating neutrophil and monocyte counts were preserved by addition of anti-C8 to the extracorporeal circuit. CONCLUSIONS: The membrane attack complex, C5b-9, is the major complement determinant of platelet activation during extracorporeal circulation, whereas C5b 9 blockade has little effect on neutrophil activation. These data also suggest a role for platelet activation or C5b-9 (or both) in the loss of monocytes and neutrophils to the extracorporeal circuit. PMID- 10469961 TI - Age-related effects of St Thomas' Hospital cardioplegic solution on isolated cardiomyocyte cell volume. AB - OBJECTIVES: We tested the hypothesis that neonatal cells are more sensitive to cardioplegia-induced cell swelling than more mature cells and spontaneous swelling in the absence of ischemia can be prevented by cardioplegia with a physiologic KCl product. METHODS: Cell volumes of isolated ventricular myocytes from neonatal (3-5 days), intermediate (10-13 days), and adult (>6 weeks) rabbits were measured by digital video microscopy. After equilibration in 37 degrees C physiologic solution, cells were suprafused with 37 degrees C or 9 degrees C St Thomas' Hospital solution (standard or low Cl(-)) or 9 degrees C physiologic solution followed by reperfusion with 37 degrees C physiologic solution. RESULTS: Neonatal cells swelled 16.2% +/- 1.8% (P <.01) in 37 degrees C St Thomas' Hospital solution and recovered during reperfusion, whereas more mature cells maintained constant volume. In contrast, 9 degrees C St Thomas' Hospital solution caused significant age-dependent swelling (neonatal, 16.8% +/- 1.5%; intermediate, 8.6% +/- 2.1%; adult, 5.6% +/- 1.1%). In contrast to more mature cells, neonatal cells remained significantly edematous throughout reperfusion (8.1% +/- 1.5%). Swelling was not due to hypothermia because 9 degrees C physiologic solution did not affect volume. Lowering the KCl product of St Thomas' Hospital solution by partially replacing Cl(-) with an impermeant anion prevented cellular edema in all groups. CONCLUSION: In the absence of ischemia, neonatal cells were more sensitive to cardioplegia-induced cellular edema than more mature cells, and edema observed in all groups was avoided by decreasing the KCl product of St Thomas' Hospital solution to the physiologic range. Differences in cell volume regulation may explain the sensitivity of neonatal hearts to hyperkalemic cardioplegic arrest and suggest novel approaches to improving myocardial protection. PMID- 10469963 TI - Commentary PMID- 10469962 TI - Emergency thoracoabdominal aortic aneurysm repair: clinical outcome. AB - OBJECTIVE: Emergency repair of thoracoabdominal aortic aneurysm remains a formidable operation with high morbidity and mortality. Although advanced surgical and perioperative care techniques have reduced the risks in elective repair of these aneurysms, the mortality rate has remained high when emergency surgery is performed. We have evaluated the outcome of patients undergoing emergency repair of thoracoabdominal aortic aneurysm. METHODS: Of 47 consecutive patients with thoracoabdominal aortic aneurysm observed from January 1993 to September 1998, 19 required an emergency operation. Twelve had a ruptured aneurysm and 7 an acute dissection. Twelve type I, 3 type II, 1 type III, and 3 type IV thoracoabdominal aortic aneurysms (Crawford's classification) were diagnosed. All patients were operated on via a thoracolaparotomy with partial femoral-femoral extracorporeal circulation. The cerebrospinal fluid pressure was monitored, and the aorta was replaced with a vascular graft. Patent intercostal arteries were reimplanted when feasible. RESULTS: The early (30-day) mortality was 42.1%; there were 2 late deaths. Complications in the 11 surviving patients are summarized as follows: paraplegia/paraparesis, 3 cases; renal, 4 cases; pulmonary, 4 cases; cardiac, 1 case; cerebrovascular, 1 case; and reexploration for bleeding, 1 case. Hemodialysis and aortic dissection were predictive factors of hospital mortality. CONCLUSIONS: Our surgical experience in emergency repair of thoracoabdominal aortic aneurysm must be considered encouraging in terms of late outcome despite the in-hospital mortality of 42.1% and serious postoperative complications in the surviving patients. Moreover, our results show that acute aortic dissection and the need for hemodialysis are predictive factors for mortality. PMID- 10469965 TI - Commentary PMID- 10469964 TI - Pulmonary autograft: should it be used in young patients with rheumatic disease? AB - BACKGROUND: Although pulmonary autograft is being increasingly used to replace the diseased aortic valve with excellent long-term results, its use in the population with rheumatic disease still needs careful evaluation. PATIENTS AND METHODS: From October 1993 through March 1998, 102 patients underwent aortic valve replacement with a pulmonary autograft (Ross procedure). The mean age was 27.9 +/- 4.2 years (range, 0.8-56 years). The cause was rheumatic disease in 75 patients (73%), bicuspid aortic valve in 26 patients (26%), and myxomatous aortoarteritis in 1 patient (1%). The root replacement technique was used in all. In addition, 31 patients had 33 associated procedures: mitral valve repair (n = 15 patients), open mitral commissurotomy (n = 15 patients), tricuspid repair (n = 2 patients), and homograft mitral valve replacement (n = 1 patient). RESULTS: Operative mortality was 6.9% (7 patients). Late mortality was 7.8% (8 patients). Follow-up ranged from 1 to 60 months (mean, 25.3 +/- 15.4 months) and was 98% complete. Two patients required reoperation for failed mitral valve repair, and 2 other patients underwent reoperation for failure of both the autograft and mitral valve repair. Echocardiographic assessment showed moderate to severe aortic regurgitation in 13 patients, along with thickening of the autograft. All of these patients had rheumatic disease and were young (<30 years). Ten of these patients had undergone associated mitral valve procedure. Morphologic and histopathologic examination of explanted autografts showed features compatible with rheumatic valvulitis. CONCLUSION: Pulmonary autograft is susceptible to rheumatic involvement. Young age (<30 years) and associated mitral valve disease are significant risk factors for autograft failure in patients with rheumatic disease. Use of pulmonary autograft in this subgroup of patients requires a cautious approach. PMID- 10469966 TI - Effect of posterior pericardiotomy on postoperative supraventricular arrhythmias and late pericardial effusion (posterior pericardiotomy). AB - OBJECTIVE: The aim of this prospective study was to evaluate the effectiveness of posterior pericardiotomy from the point of pericardial effusion related with supraventricular tachycardia and development of delayed posterior cardiac effusions. MATERIALS AND METHODS: This prospective randomized study was carried out in 200 patients undergoing coronary artery bypass surgery in Gulhane Medical Academy Department of Cardiovascular Surgery between June 1996 and June 1997. Patients were divided into 2 groups; each group included 100 patients. Longitudinal incision was made parallel and posterior to the left phrenic nerve, extending from the left inferior pulmonary vein to the diaphragm in group I patients. Posterior pericardiotomy was not done in group II. RESULTS: Atrial fibrillation was developed in 6 patients (6%) in group I and in 34 patients (34%) in group II (P =.0000007). Atrial flutter and other supraventricular arrhythmia prevalence was not statistically significant. Early and late pericardial effusion were developed 54% and 21%, respectively, in group II, but neither early nor late pericardial effusion were developed in group I (P =.00001). Delayed pericardial tamponade was also significantly lower in group I (0% vs 10%; P =.001). CONCLUSION: Posterior pericardiotomy is technically easy to perform and a safe and effective technique that reduces not only the prevalence of early pericardial effusion and related atrial fibrillation but also delayed posterior pericardial effusion and tamponade. PMID- 10469967 TI - Assessment of sternal vascularity with single photon emission computed tomography after harvesting of the internal thoracic artery. AB - OBJECTIVE: This study prospectively evaluates the effect on sternal vascularity of harvesting the left internal thoracic artery. METHODS: Twenty-four consecutive patients undergoing primary coronary artery bypass grafting were studied. One patient's procedure was altered during the operation, and he was eliminated from the study. The patients were prospectively randomized to receive a skeletonized internal thoracic artery (group I, n = 11) or a pedicled internal thoracic artery (group II, n = 12) graft. Each patient underwent a preoperative technetium 99 methylene diphosphonate bone scan using single photon emission computed tomography. The ratio of the mean counts per pixel on the left side of the sternum was compared with the mean counts per pixel on the right side. Postoperatively, all patients had a second scan, and sternal uptake was compared with the preoperative uptake. RESULTS: No significant differences in preoperative and operative variables were observed between the groups. A statistically significant reduction in blood flow to the left side of the sternum was shown postoperatively in group II compared with group I (0.61 +/- 0.11 vs 0.85 +/- 0.09; P <.001). Multivariable logistic regression analysis of preoperative and operative variables revealed only a pedicled left internal thoracic artery to be associated with a 20% or more reduction in left-to-right sternal activity ratio (odds ratio, 100; 70% confidence limits, 22-465; P =.002). CONCLUSION: A pedicled left internal thoracic artery graft to the left anterior descending artery reduces blood flow to the left side of the sternum during the acute postoperative period. This does not occur when the left internal thoracic artery is skeletonized. PMID- 10469968 TI - Extended aortic root replacement with aortic allografts or pulmonary autografts in children. AB - OBJECTIVES: To evaluate the early results and effectiveness of left ventricular outflow tract enlargement with aortic allograft or pulmonary autograft in children with complex left ventricular outflow tract obstruction. METHOD: The records of 30 children who underwent aortic root enlargement and replacement with either an aortic allograft (22 patients) or pulmonary autograft (8 patients) between January 1987 and June 1997 were reviewed. The predominant diagnosis was complex left ventricular outflow tract obstruction (n = 19), associated with aortic incompetence in 11 children. Before root enlargement, 27 children underwent surgical valvotomy (14 patients), balloon dilatation (10 patients), or both interventions (3 patients). Mean age at root enlargement was 5.4 +/- 3.5 years (range, 2 days-16 years). Most of the children (27 patients) underwent a Konno aortoventriculoplasty. Concomitant septal myectomy was performed in 4 children, mitral valve procedure in 5 children, and endocardial fibroelastosis resection in 1 child. RESULTS: Five children (17%) died in hospital. Four of these were infants less than 2 months old. All had acute aortic incompetence as the result of recent intervention necessitating urgent operation. The fifth child, aged 10 years, died of myocardial failure 2 weeks after the operation. During the follow-up period (mean length, 4.1 +/- 2.8 years), sudden death occurred in 1 child 3 months after the operation. Follow-up echocardiograms (obtained for 23 of the surviving 24 children within 3 +/- 2.3 years) showed a left ventricular outflow tract gradient reduced from a mean of 65 to 11 mm Hg (P =.001); Z value increased from a mean of -0.5 to 4.1 (P <. 001), and aortic incompetence was trivial or mild except in 2 children. CONCLUSION: Urgent aortic root enlargement in decompensating neonates carries higher mortality rates. In older children, the early results of root enlargement and implantation of allograft or autograft are good. PMID- 10469969 TI - Impact of diaphragmatic paralysis after cardiothoracic surgery in children. AB - OBJECTIVES: We sought to determine the prevalence and clinical impact of diaphragmatic paralysis caused by phrenic nerve injury after cardiothoracic surgery in children. METHODS: A search of cardiology, radiology, and hospital databases identified 170 episodes of diaphragmatic paralysis after cardiothoracic surgery in 168 children operated on from 1985 to 1997. Medical records were reviewed to determine demographics, details of the operation and postoperative course, diagnostic features and management of diaphragmatic paralysis, and follow up status. RESULTS: The prevalence of diaphragmatic paralysis was 1.6% (95% confidence interval 1.4%-1.8%). Median age at operation was 6 months (range <1 day-14.4 years). Median time from the operation to the initial investigation was 5 days (range <1 day-61 days), with 57% of patients receiving mechanical ventilation at diagnosis. Diaphragmatic plication was performed in 40% of the patients at a median interval from the initial investigation of 15 days (range 3 days-11.1 months). Significant independent factors associated with increased postoperative hospital stay were lower patient weight at operation, previous cardiothoracic operations, bilateral diaphragmatic paralysis, increased interval from operation to investigation, mechanical ventilation at the time of investigation, and diaphragmatic plication. Confirmed recovery of diaphragmatic function was noted before hospital discharge in only 15 episodes. CONCLUSIONS: Diaphragmatic paralysis complicating cardiothoracic surgery continues to occur in the current era, with a significant impact on morbidity. Smaller patients with bilateral hemidiaphragmatic paralysis, requiring mechanical ventilation, may represent a higher risk subgroup to target for increased diagnostic suspicion and more aggressive management; early spontaneous recovery is rare. PMID- 10469970 TI - Rationale and design of the National Emphysema Treatment Trial (NETT): A prospective randomized trial of lung volume reduction surgery. PMID- 10469971 TI - Independent prognostic role of p16 expression in lung cancer. AB - OBJECTIVES: The cyclin-dependent kinase p16 (also known as Ink4A, Mts1, Cdkn2, and Cdkn4i) has been proposed as a tumor suppressor gene mapped on chromosome segment 9p21. This study evaluated p16 protein expression in 135 lung cancer specimens and investigated potential genetic alterations occurring in this gene. RESULTS: We found altered p16 immunohistochemical expression to be a frequent event in lung cancer and to be independent of either the histologic type or any other clinical-pathologic feature. Western blot analyses performed on about one third of the specimens correlated highly with these results. In addition, we found p16 immunohistochemical expression to be a favorable prognostic factor in lung cancer in that its reduction or loss correlated with a worse outcome for the patients. Polymerase chain reaction amplification and direct sequencing of p16 exons 1 and 2 revealed no mutations, indicating that p16-altered expression in lung cancer is not necessarily linked to mutational events of these genes. CONCLUSIONS: We conclude that p16-altered expression is both an independent and frequent event in lung cancer and may have an important role in tumorigenesis and in malignant progression of a significant proportion of these cancers. However, the actual incidence and relevance of p16 mutations in this neoplasm continues to be debated, and its analysis seems inconclusive. Our results suggest a prognostic role for the immunodetection of this protein on formalin-fixed and paraffin embedded specimens. They further suggest its routine use in the evaluation of the frequently unpredictable behavior of lung cancer. PMID- 10469972 TI - Pulmonary function after segmentectomy for small peripheral carcinoma of the lung. AB - OBJECTIVE: The aim of this study is to compare the pulmonary function after a segmentectomy with that after a lobectomy for small peripheral carcinoma of the lung. PATIENTS AND METHODS: Between 1993 and 1996, segmentectomy and lobectomy were performed on 48 and 133 good-risk patients, respectively. Lymph node metastases were detected after the operation in 6 and 24 patients of the segmentectomy and lobectomy groups, respectively. For bias reduction in comparison with a nonrandomized control group, we paired 40 segmentectomy patients with 40 lobectomy patients using nearest available matching method on the estimated propensity score. RESULTS: Twelve months after the operation, the segmentectomy and lobectomy groups had forced vital capacities of 2.67 +/- 0.73 L (mean +/- standard deviation) and 2.57 +/- 0.59 L, which were calculated to be 94.9% +/- 10.6% and 91.0% +/- 13.2% of the preoperative values (P =.14), respectively. The segmentectomy and lobectomy groups had postoperative 1-second forced expiratory volumes of 1.99 +/- 0.63 L and 1.95 +/- 0.49 L, which were calculated to be 93.3% +/- 10.3% and 87.3% +/- 14.0% of the preoperative values, respectively (P =.03). The multiple linear regression analysis showed that the alternative of segmentectomy or lobectomy was not a determinant for postoperative forced vital capacity but did affect postoperative 1-second forced expiratory volume. CONCLUSION: Pulmonary function after a segmentectomy for a good-risk patient is slightly better than that after a lobectomy. However, segmentectomy should be still the surgical procedure for only poor-risk patients because of the difficulty in excluding patients with metastatic lymph nodes from the candidates for the procedure. PMID- 10469973 TI - Dobutamine stress echocardiography for the preoperative evaluation of patients undergoing lung volume reduction surgery. AB - BACKGROUND: Lung volume reduction surgery has been proposed as a bridge to lung transplantation and as definitive therapy for advanced chronic obstructive lung disease. However, patient selection criteria and optimal preoperative assessment have not been clearly defined. OBJECTIVE: We investigated the feasibility, safety, and value of dobutamine stress echocardiography as a predictor of major early cardiac events in patients who underwent lung volume reduction surgery. METHODS: The study population consisted of 46 patients (21 men and 25 women, mean age 59 +/- 9 years) who underwent dobutamine stress echocardiography (maximum dose 40 microg. kg(-1). min(-1) plus atropine if needed) 180 days or less before lung volume reduction surgery. Adverse cardiac events were prospectively defined and tabulated during hospitalization after the operation and at subsequent outpatient visits. RESULTS: Dobutamine stress echocardiography was interpretable in 45 of 46 (98%) patients. There were no adverse events during testing. The studies revealed normal left ventricular systolic function at rest in all patients and normal right ventricular function in all patients but one. Thirteen patients had right ventricular enlargement. Estimated right ventricular systolic pressure was mildly elevated (>40 mm Hg) in 5 patients. Four patients (9%) had stress tests positive for ischemia. There were no perioperative deaths. Follow-up was available for 44 of 45 patients at a duration of 20.0 +/- 7.0 months. Two major adverse cardiac events occurred in the same patient in whom the results of dobutamine stress echocardiography were positive for ischemia (positive predictive value 25%, 95% confidence interval 0% to 83%; negative predictive value 100%, 95% confidence interval 90 to 100%). CONCLUSION: Despite end-stage chronic obstructive lung disease and poor ultrasound windows, dobutamine stress echocardiography is feasible and safe in patients undergoing evaluation for lung volume reduction surgery. It yields important information on right and left ventricular function and has an excellent negative predictive value for early and late adverse cardiac events. PMID- 10469974 TI - Nitroglycerin as a nitric oxide donor accelerates lipid peroxidation but preserves ventricular function in a canine model of orthotopic heart transplantation. AB - BACKGROUND: Nitric oxide has cardioprotective effects through several mechanisms. However, nitric oxide may have deleterious effects in the presence of superoxide because it is converted to peroxynitrite, which then initiates lipid peroxidation. Using a canine model of orthotopic heart transplantation, we examined whether adding an organic nitric oxide donor, nitroglycerin, to preservation solution elicits lipid peroxidation after reperfusion and causes deleterious effects on coronary endothelial function and left ventricular function. METHODS AND RESULTS: The donor heart was preserved for 24 hours in cold University of Wisconsin solution with nitroglycerin (0.1 mg/mL) supplementation (group NTG, n = 8) or in standard University of Wisconsin solution (group C, n = 8). After reperfusion, changes of coronary resistance were measured during the infusion of acetylcholine (0.1 mg/min) and of sodium nitroprusside (1 mg/min), and percent coronary relaxation was calculated. Left ventricular function was evaluated by pressure-volume relations with the use of a conductance catheter, thereby deriving the slopes of end-systolic pressure-volume relation, stroke work end-diastolic volume relation, and maximum rate of change of left ventricular pressure-end-diastolic volume relation. Serum lipid peroxide level was measured. Percent coronary relaxation was similar for the 2 groups. The slopes of end systolic pressure-volume relation, stroke work-end-diastolic volume relation, and maximum rate of change of left ventricular pressure-end-diastolic volume relation in group NTG were significantly higher than those in group C. On the other side, serum lipid peroxide level in group NTG was significantly higher than that in group C. CONCLUSIONS: Nitroglycerin may have detrimental effects evidenced by the increase in lipid peroxidation, which implied peroxynitrite formation. However, the overall effect of nitroglycerin was cardioprotective. Although the exact mechanism is yet to be clarified, the superb cardioprotective effect of nitroglycerin overwhelms the exaggeration of lipid peroxidation. PMID- 10469975 TI - Carbon dioxide embolism during endoscopic saphenectomy for coronary artery bypass surgery. PMID- 10469976 TI - Total body retrograde perfusion during operations on the descending thoracic aorta. PMID- 10469977 TI - Thymoma encasing last patent vein graft to the heart. PMID- 10469978 TI - Externally supported radial artery graft for myocardial revascularization: A new technique to avoid vasospasm. PMID- 10469979 TI - Symptomatic coronary-subclavian steal syndrome: report of a case with complete occlusion of proximal left subclavian artery and anomalous origin of left vertebral artery from the aortic arch. PMID- 10469980 TI - Fourier analysis of the intra-aortic balloon pump. PMID- 10469981 TI - Is alpha-stat management still justified for deep hypothermic circulatory arrest in adults? PMID- 10469982 TI - Is alpha-stat management still justified for deep hypothermic circulatory arrest in adults? PMID- 10469983 TI - Introduction to the proceedings of the first symposium on elasmobranch endocrinology, held at the thirteenth annual meeting of the american elasmobranch society (AES&rpar PMID- 10469984 TI - Unique evolution of neurohypophysial hormones in cartilaginous fishes: possible implications for urea-based osmoregulation. AB - Most bony vertebrate species display a great evolutionary stability of their two neurohypophysial hormones, so that two molecular lineages, isotocin-mesotocin oxytocin and vasotocin-vasopressin, have been traced from bony fishes to mammals. Chondrichthyes, in contrast, show a striking diversity of their oxytocin-like hormones, yet show a substantial decrease in vasotocin stored in neurohypophysis when compared to nonmammalian bony vertebrates. In the rays, glumitocin ([Ser(4),Gln(8)]-oxytocin) has been identified. In the spiny dogfish, aspargtocin ([Asn4]-oxytocin) and valitocin ([Val(8)]-oxytocin) have been characterized whereas in the spotted dogfish, asvatocin ([Asn(4),Val(8)]-oxytocin) and phasvatocin ([Phe(3),Asn(4),Val(8)]-oxytocin) have been found. Finally, in the holocephalian Pacific ratfish, oxytocin, the typical peptide of placental mammals, has been discovered. The duplication of the oxytocin-like hormone gene found in dogfishes has been observed only in some Australian and American marsupials. Cartilaginous fishes have developed an original urea-based osmoregulation involving a glutamine-dependent urea synthesis and blood urea retention through renal urea transporters. Furthermore, marine species use a rectal salt gland for sodium chloride excretion. Although vasopressin, in mammals, and vasotocin, in nonmammalian tetrapods, are clearly implied in water and salt homeostasis, the hormones involved in the blood osmotic pressure regulation of elasmobranchs are still largely unknown. It is suggested that the great diversity of oxytocin-like hormones in elasmobranchs expresses a release from an evolutionary receptor-binding constraint, so that amino-acid substitutions reflect neutral evolution. In contrast, the preservation of vasotocin suggests a selective pressure, which may be related to the regulation of renal urea transporter-recruitment mechanisms, as it has been shown for vasopressin in mammals. J. Exp. Zool. 284:475-484, 1999. PMID- 10469985 TI - Elasmobranch color change: A short review and novel data on hormone regulation. AB - Skins of Potamotrygon reticulatus are light in color in vitro, exhibiting punctate melanophores. Alpha-Melanocyte stimulating hormone (EC(50) = 4.58 x 10( 9) M) and prolactin (EC(50) = 1.44 x 10(-9) M) darken the skins in a dose dependent manner. The endothelins ET-1, ET-2 and ET-3, and the purines, ATP, and uracil triphosphate (UTP) were not able to induce either skin lightening or darkening. Forskolin and the calcium ionophore A23187 promoted a dose-dependent darkening response, whereas N(2), 2'-O-dibutyryl guanosine 3'-5'-cyclic monophosphate (db cyclic GMP), phorbol-12-myristate-13-acetate (TPA), and 1 oleoyl-2-acetyl-sn-glycerol (OAG) were ineffective. The maximal response obtained with the calcium ionophore A23187 was only 76% of maximal darkening. These results indicate that the cyclic adenosine 3'-5'-monophosphate (cAMP) pathway is probably involved in the pigment dispersion of P. reticulatus melanophores. Other experiments should be done to further investigate how cytosolic calcium may be physiologically increased, and the existence of a putative cross-talk between calcium and cAMP signals. In conclusion, the only hormones effective on P. reticulatus melanophores were prolactin and alpha-MSH. No aggregating agent has been shown to antagonize these actions. Prolactin effect on elasmobranch melanophores adds a novel physiological role to this ancient hormone. J. Exp. Zool. 284:485-491, 1999. PMID- 10469986 TI - Evaluation of thyroid hormone economy in elasmobranch fishes, with measurements of hepatic 5'-monodeiodinase activity in wild dogfish. AB - This paper reviews the current understanding of thyroid hormone economy and homeostasis in elasmobranch fishes and considers those measures of the activity of the hypothalamic-pituitary gland-thyroid gland-peripheral tissue axis that are necessary for adequate assessment of thyroid hormone physiology. In particular, we focus on the value of measuring hepatic 5'-monodeiodinase (5'-MDA) activity as an indicator of the animal's cellular production rate of the active thyroid hormone, triiodo-L-thyronine (T(3)). We also examine the characteristics of hepatic 5'-MDA activity, in vitro, in adult female dogfish (Squalus acanthias) collected from Passamaquoddy Bay, New Brunswick, Canada, and in the embryos that they were carrying. T(3) production from T(4) by hepatic homogenates in vitro was time- and temperature-dependent, and was enhanced by the presence of a thiol donor. Michaelis constant (K(m)) and maximum reaction velocity (V(max)) values were 3.8 x 10(-7) M and 0.29 nM T(3)/mg protein/hr, respectively. The inclusion of trimethylamine-N-oxide (TMAO) or a mixture of urea, TMAO, betaine and sarcosine significantly enhanced T(3) production. Hepatic 5'-MDA activity was depressed in fish fasted for 7 days. J. Exp. Zool. 284:492-499, 1999. PMID- 10469987 TI - Serum T(4) and serum T(3) concentrations in immature captive whitetip reef sharks, Triaenodon obesus. AB - Serum T(3) (3,5,3' triiodothyronine) and serum T(4) (thyroxine) concentrations were repetitively assayed by radioimmunoassay over a three-year period in two male and two female immature captive whitetip reef sharks, Triaenodon obesus. These sharks were maintained at the Waikiki Aquarium, Honolulu, Hawaii, in an open system holding pool receiving 568 liters per minute of water from a saltwater well with an iodide concentration of 0.076 mg/liter. No significant male-female difference was observed for either serum T(3) or serum T(4). No seasonal pattern of serum T(3) was detected (P = 0.07). Serum T(3) concentrations ranged (mean +/- SEM) from 0. 52 to 0.83 ng/mL (0.67 +/- 0.01; n = 64). A significant seasonal difference was observed for serum T(4) (P < 0.001). Serum T(4) concentration was higher in winter (October-January) with a mean (range +/- SEM) of 6.58 ng/mL (1.48-8.77 +/- 0.35; n = 24) and lower in summer (May-August) with a mean of 3.62 ng/mL (1.34-5.71 +/- 0. 22; n = 24). The thyroid hormone T(4) has a seasonal rhythm even in immature sharks and may have an important role in physiology. J. Exp. Zool. 284:500-504, 1999. PMID- 10469988 TI - Structure of the thyroid gland, serum thyroid hormones, and the reproductive cycle of the Atlantic stingray, Dasyatis sabina. AB - This study examines the role of the thyroid gland in the control of reproduction in the viviparous Atlantic stingray, Dasyatis sabina. Thyroid activity in individuals in different reproductive stages was assessed both by microscopic examination of the gland, and by analysis of circulating levels of thyroid hormones from the same individuals. The thyroid gland is a cylindric organ, embedded in a connective tissue capsule, and composed of follicles, i.e., monolayer spheres of thyroid epithelial cells. Stingray follicular cells possess several characteristic features, namely apical cilia and a well-developed endoplasmic reticulum. Cells vary in size and shape, according to the activity of the gland. No structural differences were observed between the thyroid glands of the two sexes. Both thyroid hormones, triiodothyronine, [T(3)], and thyroxine, [T(4)], were detected in the serum of all animals examined. Levels ranged from 1.3-2.6 microg/100 ml for total T(4), and from 1.2-2.6 ng/ml for total T(3). The T(4) levels did not vary significantly in any group. Immature individuals and females undergoing oogenesis had the lowest levels of circulating T(3) and mature females from ovulation throughout gestation had high thyroid gland activity and high levels of circulating T(3). J. Exp. Zool. 284:505-516, 1999. PMID- 10469989 TI - Regulation of interrenal gland steroidogenesis in the Atlantic stingray (Dasyatis sabina). AB - The interrenal gland (the homologue of the mammalian adrenal cortex) of elasmobranchs (sharks, skates, and rays) produces 1alpha-hydroxycorticosterone (1alpha-B), which has been reported to function both as a gluco- and mineralocorticosteroid. In vitro synthesis of 1alpha-B by Atlantic stingray (Dasyatis sabina) interrenal glands was stimulated by short-term (2 hr) and long term (24 hr) treatment with porcine adrenocorticotropic hormone (pACTH). Cycloheximide blocked the pACTH-induced effect on 1alpha-B synthesis, thus demonstrating that the mechanism for the short-term induction of steroidogenesis involved protein synthesis. However, gene transcription did not play a role in the short-term induction of 1alpha-B synthesis, as indicated by the lack of an effect with actinomycin D treatment. Long-term in vitro exposure to pACTH (but not short-term exposure) stimulated the synthesis of another steroid, 11 dehydrocorticosterone (A). This induction was partially blocked by cycloheximide and actinomycin D, which suggests enhanced expression of the 11beta hydroxysteroid dehydrogenase gene. In addition, the 24-hr treatment with pACTH enhanced the activity of cytochrome P450 side chain cleavage several fold and doubled the activity of 3beta-hydroxysteroid dehydrogenase and cytochrome P450 21 hydroxylase in D. sabina interrenals, again suggesting the induction of steroidogenic genes. In contrast to other elasmobranch species, the salmon and human forms of angiotensin II had no effect on D. sabina interrenal steroidogenesis. J. Exp. Zool. 284:517-525, 1999. PMID- 10469990 TI - Renin-angiotensin system in elasmobranch fish: A review. AB - The renin-angiotensin system (RAS) has been identified recently in elasmobranch fish, and the structure of angiotensin II (ANG II) is unusual ([Asp(1),Pro(3),Ile(5)]-ANG II) compared to other vertebrates. Receptors for ANG II have been identified in blood vessels and in a variety of osmoregulatory tissues including the gill, kidney and rectal gland. In addition, there is considerable binding to the interrenal gland and the stimulation of 1alpha hydroxycorticosterone production in vitro suggests a physiological role in corticosteroidogenesis. ANG II is a potent vasoconstrictor and this effect does not appear to be mediated by sympathetic activation or catecholamine release. Although the RAS may not be involved in maintaining basal blood pressure, it may be important in situations in which blood pressure is reduced. Understanding of the role of ANG II as an osmoregulatory hormone is only just emerging with putative roles in the control of gill, rectal gland and perhaps, drinking. In addition, the stimulation of corticosteroid secretion may provide another means of controlling osmoregulation. J. Exp. Zool. 284:526-534, 1999. PMID- 10469991 TI - Molecular diversity, localization, and biological actions of elasmobranch tachykinins. PMID- 10469992 TI - Parathyroid hormone-related protein (PTHrP) in cartilaginous and bony fish tissues. AB - Tissues from a range of fish were examined for the presence of parathyroid hormone-related protein (PTHrP) to investigate PTHrP protein distribution and PTHrP gene expression in jawless fish, cartilaginous fish, and bony fish. Immunoreactive PTHrP was localized using antisera to N-terminal and mid-molecule regions of human PTHrP and PTHrP gene expression examined using a digoxigenin labeled riboprobe to a conserved region of the mammalian PTHrP gene. In all of the fish studied, PTHrP protein and messenger RNA (mRNA) were localized to the skin, kidney, and skeletal muscle, following the pattern seen in higher vertebrates. Additional sites of localization for both protein and mRNA included gill, nerve cord, and pituitary, as well as developing dermal denticles and rectal gland in the elasmobranch species. The sites of PTHrP distribution indicate that PTHrP may have roles in ionoregulation as well as growth and differentiation in fish, as has been suggested in higher vertebrates. The results imply that the distribution of PTHrP is widespread in fish and that there is homology between the PTHrP molecules found in humans and fish. The conservation of localization and possible similarity of the PTHrP molecules between tetrapods and fish suggests that PTHrP has a number of fundamental roles in vertebrates. J. Exp. Zool. 284:541-548, 1999. PMID- 10469993 TI - Effects of insulin-like growth factor-I, corticosterone, and 3,3', 5-tri-iodo-L thyronine on glycosaminoglycan synthesis in vertebral cartilage of the clearnose skate, Raja eglanteria. AB - Effects of insulin-like growth factor-I (IGF-I), corticosterone, and triiodothyronine (T(3)) on in vitro growth of vertebral cartilage of the clearnose skate, Raja eglanteria, were investigated. Uptake of [(35)S]sulfate in cultured vertebrae was used to characterize glycosaminoglycan (GAG) synthesis and cartilage growth. IGF-I significantly enhanced cartilage growth when concentrations of 1.28 and 12.8 nM were present in the culture system. Corticosterone significantly inhibited vertebral GAG synthesis at concentrations of 1, 10, and 100 nM. This effect was markedly pronounced in cartilage exposed to 1 and 10 nM corticosterone, in which GAG synthesis was virtually ceased. In contrast, T(3) (0.75, 7.5, and 75.0 nM) had no significant effect on sulfate uptake. These data suggest that IGF-I and corticosteroids may play important roles in regulating skeletal growth of elasmobranchs, as they appear to do in other vertebrates. While T(3) does not appear to exert an immediate, direct effect on vertebral growth, it may still influence elasmobranch chondrogenesis over longer culture periods or indirectly through other regulatory pathways. Thus, further information is necessary to characterize the role of thyroid hormones in the skeletal growth of these fishes. The present study is the first in vitro investigation on the hormonal regulation of elasmobranch cartilage growth. As such, the methods described herein provide a useful technique for examining these physiological processes. J. Exp. Zool. 284:549-556, 1999. PMID- 10469994 TI - Reproductive endocrinology of female elasmobranchs: lessons from the little skate (Raja erinacea) and spiny dogfish (Squalus acanthias). AB - Conventional classification of reproductive modes in female elasmobranchs fails to account for the diversity in ovarian dynamics that operate during oviparous and viviparous cycles. Delineating this diversity is crucial for understanding the endocrine regulation of the manifold physiological mechanisms utilized to retain and protect eggs and developing embryos, to fuel embryogenesis, and to manage the intrauterine milieu. Oocyte development and follicular steroidogenesis overlap with egg retention and pregnancy in some species, whereas in others the follicular phase of the cycle is temporally separated from the gravid period. A luteal phase predominates the post-ovulatory period in viviparous species. In oviparous species, the luteal phase overlaps with the follicular cycle. This heterogeneity in ovulatory cycles suggests that the endocrine system evolved a transmutable system for regulating steroidogenesis and the control of the reproductive events. The reproductive biology and endocrinology of the oviparous little skate and lecithotrophic viviparous spiny dogfish are reviewed in order to derive a working hypothesis that explains the complex nature of endocrine patterns observed in species utilizing disparate reproductive modes. An understanding of the adaptations in ovarian dynamics to particular ovulatory cycles is key to developing theories about the evolution of reproductive strategies in female elasmobranchs. J. Exp. Zool. 284:557-574, 1999. PMID- 10469995 TI - Alterations in serum steroid concentrations in the clearnose skate, Raja eglanteria: correlations with season and reproductive status. AB - Serum steroid hormones in the peripheral circulation of the clearnose skate, Raja eglanteria, were measured at the time of capture and at various times throughout the year while the animals were maintained as a captive breeding population. These analyses demonstrate interesting correlations between changes in hormone concentrations and annual reproductive events. Animals were sampled once (78 females, 20 males) or multiple times (15 females). For both groups of females, 17beta-estradiol was detected throughout the year with significant elevations occurring during October and November when ovarian follicles begin to mature (as determined through necropsy examinations), and January and February when maximum mating activity is observed and egg laying begins. Testosterone and dihydrotestosterone concentrations were significantly elevated in females only during January and February. Testosterone elevations were synchronous with longer term elevations in 17beta-estradiol in females sampled either once or repetitively. Testosterone concentrations in males were significantly elevated during times of maximum breeding activity compared to periods of sexual inactivity. Data from females sampled during five stages of the egg laying process, as defined by the position of palpable egg capsules within the reproductive tract, revealed that 17beta-estradiol was highest when egg capsules were forming in the nidamental gland (stage 2) or uterus (stage 3); testosterone and dihydrotestosterone were maximal when eggs were in the uterus (stage 3) or cloaca (stage 4); and progesterone was significantly elevated immediately after oviposition (stage 5), suggesting a possible role for progesterone in the regulation of sequential laying of egg pairs. J. Exp. Zool. 284:575-585, 1999. PMID- 10469996 TI - Plasma steroid hormone profiles and reproductive biology of the epaulette shark, Hemiscyllium ocellatum. AB - Examination of the reproductive biology of the oviparous epaulette shark, Hemiscyllium ocellatum, was conducted on a wild population. Male sharks were found to reach maturity at between 55-60 cm total length (TL) and female sharks mature around 55 cm TL. Blood samples collected from mature male and female sharks were analyzed for sex steroid hormones to examine seasonal hormone patterns. Plasma samples were analyzed via radioimmunoassay techniques with female samples measured for estradiol, progesterone, and androgen concentrations, and male samples measured for androgen concentrations. Male androgen concentrations showed a single broad peak from July to October with maximum hormone concentrations (60 ng/ml) occurring in August. Male androgen concentrations were lowest in December-February (<20 ng/ml), and appeared to correlate with reproductive activity and water temperature. Female androgen concentrations were an order of magnitude lower than those for males and showed peaks in June (6 ng/ml) and December (8 ng/ml). Estradiol concentrations in females peaked during the months of September-November (0.5 ng/ml) coinciding with the egg laying period. Progesterone concentrations ranged up to 0.5 ng/ml prior to the mating season. Observations of ova size and egg production showed eggs develop in pairs and ova are ovulated at a size of 25-27 mm. Females lay eggs from August to January. Males were observed with swollen claspers from July through December, with the highest amount of sperm storage in the epididymis occurring between August through November. Our observations indicate that epaulette sharks in the waters near Heron Island mate from July through December. J. Exp. Zool. 284:586-594, 1999. PMID- 10469997 TI - Serum steroid hormones including 11-ketotestosterone, 11-ketoandrostenedione, and dihydroprogesterone in juvenile and adult bonnethead sharks, Sphyrna tiburo. AB - Previous studies in the placental viviparous bonnethead shark, Sphyrna tiburo, have correlated 17 beta-estradiol, progesterone, testosterone, and dihydrotestosterone with reproductive events in both males and females. However, several key reproductive events, including implantation, maintenance of pregnancy, and parturition, did not correlate with these four steroid hormones. Therefore, the present study investigated three steroid hormones, 11 ketotestosterone, 11-ketoandrostenedione, and dihydroprogesterone, which have demonstrably important roles in the reproductive cycles of teleosts. It was hypothesized that one or more of these three hormones would correlate with specific reproductive events in S. tiburo. Concurrently, developmental (growth and/or maturation) analyses of these three steroids plus 17 beta-estradiol, progesterone, testosterone, and dihydrotestosterone were investigated in juvenile bonnethead sharks. Serum dihydroprogesterone concentrations were highest in mature females and 11-ketotestosterone concentrations were highest in mature males. In mature females, 11-ketoandrostenedione levels were elevated from the time of mating, through six months of sperm storage and another four months of gestation. At parturition concentrations became significantly lower and remained lower until mating occurred again in another two to three months. Serum 11 ketotestosterone concentrations were the highest at implantation though not significant. In mature males, significantly elevated serum levels of dihydroprogesterone occurred in April and May, near the start of annual testicular development. During growth in males, testosterone and dihydrotestosterone increased progressively and in females, testosterone increased progressively. At maturity in males, significant increases occurred in testosterone and 11-ketotestosterone concentrations while, in females, dihydroprogesterone, 11-ketotestosterone, 17 beta-estradiol, progesterone, testosterone, and dihydrotestosterone concentrations increased. This study shows that although testosterone may be the primary androgen in the bonnethead shark, other derived androgens may have important functions in growth, maturation, and reproduction. J. Exp. Zool. 284:595-603, 1999. PMID- 10469998 TI - A study of lung cancer mortality in asbestos workers: Doll, 1955. AB - Between 1935 and 1953, a series of publications appeared in England, Germany and America reporting cases of lung cancer amongst asbestos workers. As early as 1943, the German scientific consensus was that the evidence was strong enough to deem the association to be causal. On reviewing a more extensive bibliography, this view was shared by an American cancer expert. The results of industry sponsored experiments, in which lung tumors had been induced in mice by asbestos, were circulated in confidence to its scientists, but being unpublished were unknown to the general scientific community. There were also cancer mortality data recorded for populations of exposed asbestos workers, but these were confidential and remained to be analyzed. To deal with the persistent allegations of a lung cancer hazard, in 1953 Dr. Richard Doll was asked by Turner Brothers Asbestos ("the Company"), whose in-house analyses had been reassuring, to study the mortality data of a group of its workers. Despite the limitations of the data, Doll convincingly demonstrated so substantial an excess of lung-cancer in heavily exposed long-term asbestos workers as to overcome honest doubt. Despite determined attempts made to dissuade them, Doll and the editor of the journal to which he submitted his paper, courageously went ahead and published the paper. Industry overestimated the harm that publication of the paper would do to their immediate interests. It produced so profound a lack of sense of urgency, that legislation addressed to the control of lung cancer specifically had to wait 20 years, and asbestos workers contracting it were to wait 25 years, before they might be considered for compensation, and even then, only under extremely limiting conditions. PMID- 10469999 TI - Estimating occupational radiation doses when individual dosimetry information is not available: a job exposure matrix. AB - BACKGROUND: A job exposure matrix (JEM) was developed for a population based case control study to assess the possible association between occupational radiation and non-Hodgkin's lymphoma. METHODS: Using published radiation monitoring data, we developed a radiation JEM composed of estimated annual dose distributions, categorized by time period, for a broad range of occupational and industrial groups. RESULTS: When information is available to correctly assess an individual's exposure status, the annual dose distributions in the JEM can be used in conjunction with job histories to estimate the distribution of possible cumulative doses for individuals. The median of the cumulative dose distribution can then be used in standard epidemiologic analysis. In addition, methods can be applied that incorporate the uncertainty about each individual's true dose into risk estimates and associated confidence intervals. CONCLUSIONS: The JEM can be useful in estimating occupational radiation exposures in other studies, particularly population based case control studies which include detailed occupational histories. PMID- 10470000 TI - Asbestos exposure in lung carcinoma: a necropsy-based study of 414 cases. AB - BACKGROUND: The prerequisites necessary for attributing lung carcinoma to asbestos, represent a controversial issue. METHODS: Three parameters (occupational history, pleural plaques, and lung asbestos bodies) were investigated in 414 consecutive cases of lung carcinoma, examined at necropsy at the Hospital of Monfalcone, Italy. Occupational data were obtained from the patients' relatives by personal or telephone interviews. Pleural plaques were classified into three classes (small, moderate, large). Routine lung sections were examined for asbestos bodies in all cases; isolation and counting were performed in 408 cases. RESULTS: The series included 353 men, and 61 women, aged between 38 and 97 years. The male patients had worked in industries in 74% of cases (60% in shipbuilding). Men showed pleural plaques in 82% of cases (moderate or large plaques in 58.7%). Asbestos bodies were observed in routine lung sections in 34.8%, and in 31% exceeded the value of 5,000 bodies per gram of dried tissue. Among women the principal features were: history or domestic exposure to asbestos in 36% of the cases, prevalence of pleural plaques 34% (moderate or large plaques 15%), asbestos bodies in routine lung sections in 3.3% and there was no case with an asbestos body burden over 5,000/g. The fraction of asbestos-related carcinomas among male patients varied between 24.7 and 61%, depending on the criteria used for attribution. CONCLUSIONS: Different criteria indicated about 60% of the present lung carcinomas among men as plausibly attributable to asbestos. PMID- 10470001 TI - Gambling, drinking, smoking and other health risk activities among casino employees. AB - BACKGROUND: This study examined the prevalence of level 3 (pathological) gambling and a variety of other health risks among casino employees. METHODS: A sample of 3841 full-time casino employees representing four geographic sites was surveyed about gambling, drinking, smoking, and other health risk behaviors. In addition, respondents were asked about their use of the employee assistance program (EAP) and perceived obstacles towards using the EAP. RESULTS: This study found that casino employees have a higher prevalence of past-year level 3 (pathological) gambling behavior than the general adult population, but a lower prevalence of past-year level 2 (problem) gambling than the general adult population. In addition, casino employees have higher prevalence of smoking, alcohol problems, and depression than the general adult population. Furthermore, these risk behaviors tend to cluster. The majority of non-smoking respondents in this sample were exposed to second-hand smoke. Employees reported low participation in the company's EAP. CONCLUSIONS: The results of this study suggest that casino management should consider (1) improving problem gambling screening for employees who visit EAPs, even if employees present other problems (e.g., alcohol problems) as their primary concern, (2) increasing employees' awareness of EAPs, (3) increasing health promotion and education through channels other than company EAPs, and (4) creating smoke-free working areas. PMID- 10470002 TI - Paraoxonase polymorphism and its effect on male reproductive outcomes among Chinese pesticide factory workers. AB - BACKGROUND: Serum paraoxonase has been associated with the metabolism of organophosphate pesticides in humans. Molecular analysis of the human paraoxonase gene (PON1) has revealed that Arg192 homozygotes have a greater detoxifying capability than Gln192 homozygotes. We examined the effects of PON1 genotypes on male reproductive outcomes and its interaction with exposure to organophosphate pesticides. METHODS: We studied 60 Chinese pesticide-factory workers and 89 textile-factory workers who were unexposed to pesticides. The respective allele frequencies of Arg192 and Gln192 were 0.62 and 0.38. Pesticide exposure among 36 exposed subjects and 12 unexposed subjects, regardless of gender, was assessed by personal measurement of pesticide residues over an entire 8-hr shift and measurement of urinary p-nitrophenol level over a 24-hr period. We analyzed semen and hormone data collected from male subjects. RESULTS: When the three PON1 genotypes were analyzed separately, a gene dose effect was not detected. We used the unexposed Arg192 homo/heterozygotes as the reference group, and re-analyzed the data. Exposed Arg192 homo/heterozygotes had significantly lower sperm count (chi 2 = 9.01, P < 0.01) and lower percentage of sperm with normal morphology (chi 2 = 4.18, P < 0.05) than the reference group. Both unexposed Gln192 homozygotes (chi 2 = 4.90, P < 0.05) and exposed Arg192 homo/heterozygotes (chi 2 = 10.00, P < 0.01) showed significantly lower sperm concentrations than the reference group. In addition, exposed Arg192 homo/heterozygotes had significantly higher serum LH levels (chi 2 = 7.94, P < 0.01) than the reference group. CONCLUSIONS: Because of a small sample size, our findings are highly preliminary. Nevertheless, it calls for further investigation of the interaction between the PON1 genotype and organophosphate pesticide exposure on male reproductive outcomes. PMID- 10470003 TI - Physical capacity in relation to present and past physical load at work: a study of 484 men and women aged 41 to 58 years. AB - BACKGROUND: A negative association has previously been reported between long lasting physically heavy work and some measures of physical capacity. This relationship was further investigated in a 24-year follow-up study of 484 middle aged men and women from the general population. METHODS: A questionnaire was administered in 1993 concerning retrospective recall of physical work loads and physical training in the time span between 1970 and 1993. Laboratory tests performed in 1993 included tests of muscle function (maximal isometric strength and dynamic endurance) and aerobic power (submaximal ergometer test). RESULTS: Consistent with the hypothesis, but mainly among the women, associations between long-lasting physically heavy demands and low trunk flexion strength, squatting endurance, and aerobic power were observed. In contrast, low isometric hand grip strength and low weight lifting endurance were seldom seen among those with high physical work loads, indicating a possible maintaining or training effect on the hand/arm/shoulder muscle groups. CONCLUSIONS: Physically heavy work seems to have a different impact on different parts of the musculoskeletal system, an effect that is also different between men and women. PMID- 10470005 TI - Lung cancer risk in male workers occupationally exposed to diesel motor emissions in Germany. AB - BACKGROUND: Although in several epidemiological studies exposure to diesel motor emissions (DME) shows an elevated lung cancer risk, it is still controversial whether DME is a human carcinogen. METHODS: In a pooled analysis of two case control studies on lung cancer in Germany a total of 3498 male cases with histologically or cytologically ascertained lung cancer and 3541 male population controls were included. Information about lifelong occupational and smoking history was obtained by questionnaire. Drivers of lorries, buses, taxies, diesel locomotives and forklift trucks, bulldozers, graders, excavators, and tractors, were considered as exposed to DME and their cumulative exposure was estimated. All odds ratios were adjusted for smoking and asbestos exposure. RESULTS: The evaluation of lung cancer risk for all jobs with DME-exposure combined showed an odds ratio of OR=1.43 (95%-CI: 1.23-1.67). Most pronounced was the increase in lung cancer risk in heavy equipment operators (OR=2. 31 95%-CI: 1.44-3.70). The risk of tractor drivers increased with length of employment and reached statistical significance for exposures longer than 30 years (OR=6.81, 95%-CI: 1.17-39.51). The group of professional drivers (e.g., trucks, buses, and taxies), showed an increased risk only in West Germany (OR=1.44, 95%-CI: 1. 18-1.76), but not in East Germany (OR=0.83, 95%-CI: 0.60-1.14). DME-exposure in other traffic related jobs (e.g., diesel engine locomotive drivers, switchmen, forklift operators) was associated with an odds ratio of OR=1.53 (95%-CI: 1.04-2.24). CONCLUSIONS: The study provides further evidence that occupational exposure to diesel motor emissions is associated with an increased lung cancer risk. PMID- 10470006 TI - Diesel exhaust and lung cancer mortality in potash mining. AB - BACKGROUND: Findings from experimental studies on rodents and from epidemiological studies suggest that diesel exhaust may cause lung cancer. There is evidence that in several occupations, e.g., truck drivers and railway workers, the risk of lung cancer increases with duration of employment, and exposure to diesel exhaust provides the most likely explanation for these elevations of risk. METHODS: We investigated the association between lung cancer mortality and exposure to diesel exhaust in a cohort study. The cohort comprised 5, 536 male potash miners who were followed from 1970 to 1994. Exposure was assessed from concentration measurements of the total carbon (i. e., elemental and organic carbon in total) in personal dust samples. The concentration values were multiplied by years of exposure to give a quantitative exposure measure. The concentration levels ranged from 0.12 to 0.39 mg/m(3) total carbon in fine dust. Work histories and smoking habit data were obtained from medical company records. Causes of death were ascertained from death certificates. RESULTS: During the follow-up period, 424 deaths were recorded, including 133 of cancer, 38 of lung cancer. The relative risk of lung cancer between two groups with high and low exposure was 2.2 (95% confidence interval 0.8-6.0). With Cox regression, we found a lung cancer relative risk 1.7 (0.5-5.8) after twenty years of exposure. Extensive scrutiny proved smoking not to be a confounder in this study. CONCLUSIONS: The principal finding of the study is a doubling of relative lung cancer risk after twenty years of exposure in the workplaces with highest exposure. However, the observed elevation is nonsignificant even at a 90% level. Further follow-up is intended to enhance the study power. PMID- 10470007 TI - Mortality among chemical plant workers exposed to acrylonitrile and other substances. AB - OBJECTIVES: To examine the association between exposure to acrylonitrile (AN) and cancer mortality by performing an independent and extended historical cohort study of workers from a chemical plant in Lima, Ohio included in a recent NCI NIOSH study. METHODS: Subjects were 992 white males who were employed for three or more months between 1960 and 1996. We identified 110 deaths and cause of death for 108. Worker exposures were estimated quantitatively for AN and qualitatively for nitrogen products. Statistical analyses included U.S. and local county-based SMRs and internal relative risk regression of internal cohort rates. RESULTS: No statistically significant excess mortality risks were observed among the total cohort for the cancer sites implicated in previous studies: stomach, lung, breast, prostate, brain, and hematopoietic system. We observed a statistically significant bladder cancer excess based on four deaths (SMR=7.01, 95% CI=1.91 17.96) among workers not exposed to AN. Among 518 AN-exposed workers, we observed a not statistically significant excess of lung cancer based on external (SMR=1.32, 95% CI=.60-2.51) and internal (RR=1.98, 95% CI=.60-6.90) comparisons. Although the trends were not statistically significant, exposure-response analyses of internal cohort rates showed monotonically increasing lung cancer rate ratios with increasing AN exposure, with RRs exceeding 2.0 in the highest exposure categories. CONCLUSIONS: With the possible exception of lung cancer, this study provides little evidence that exposure to AN at levels experienced by Lima plant workers is associated with an increased risk of death from any cause including the implicated cancer sites. PMID- 10470008 TI - Occupation and histopathology of lung cancer: A case-control study in Rosario, Argentina. AB - BACKGROUND: Male age-adjusted lung cancer mortality rate is high in Rosario City, Argentina (62.7 per 100,000). METHODS: Case-control study involving interviews with 367 male lung cancer patients and 576 controls. RESULTS: The histological distribution detected was: squamous cell 39%, adenocarcinoma 34%, small cell carcinoma 13%, and other or no specified cell type 14%. When comparing with the group unlikely to be exposed to occupational carcinogens, a 60% increase in risk was observed for the remaining occupations (P < 0.008). Risks were high for drivers (OR=1.9, CI: 1.1-4.0), construction (OR=2.5, CI: 1.0-5.9), and agricultural workers also (OR=1.8, CI: 1. 1-3.1). In regard to squamous cell carcinoma, increased risks were observed in the metal industry, particularly in welders (OR=2.9, CI: 1.0-10.1) and mechanics (OR=1.8, CI: 0.9-4.2). Smoking was not a substantial confounding effect. CONCLUSIONS: Occupational exposures partly account for the high lung cancer mortality rate among male residents of Rosario City. PMID- 10470009 TI - Work-related respiratory disorders and farming characteristics among cattle farmers in Northern Germany. AB - BACKGROUND: The aim of this cross-sectional study was to investigate the prevalence of respiratory symptoms among Northern German farmers in relation to cattle farming characteristics. METHODS: 1735 farmers were visited on their farms and were interviewed using a standardized questionnaire on work-related respiratory symptoms and farming details. RESULTS: 84.6% of the farmers were cattle farmers. The prevalence of work-related respiratory symptoms was 40.3%. In a multiple logistic regression model adjusting for response rate, age, gender, and smoking habits, work-related respiratory symptoms were shown to be significantly associated with the ventilation of the cattle house (OR (ventilation via the wall): 0.57), feeding management (OR (feeding once daily): 0.53), and plant crop (OR: 0. 75). Farmers, living inland, showed a significant higher prevalence of work-related respiratory symptoms (OR: 1.34). CONCLUSIONS: The use of ventilation via the wall might be recommended for new cattle houses in regions with warm winters. PMID- 10470010 TI - Case-series investigation of intracranial neoplasms at a petrochemical research facility. AB - BACKGROUND: From 1970 through 1997, 17 intracranial neoplasms were identified among 6,800 employees of a petrochemical research facility. This investigation describes the case-series. METHODS: The intracranial neoplasms were identified by self reports and record linkages, and were confirmed by medical records and a pathology review. Standardized incidence ratios (SIRs) compare observed and expected numbers of cases according to certain work characteristics. RESULTS: Overall, there were 17 observed and 10.5 expected intracranial neoplasms, including 11/4.7 benign intracranial tumors and 6/5.9 brain cancers. All brain cancers occurred among male research scientists or technicians. Four had worked at some time on the same floor of one building (SIR=12.6, 95% CI=3.4-32.1), and several had worked on a research project with at least one other brain cancer case. The benign intracranial neoplasm cases did not have common building assignments or work activities. CONCLUSIONS: The occurrence pattern of brain cancers, but not that of benign tumors, suggests a possible occupational etiology. PMID- 10470011 TI - Current diagnostic methods for diisocyanate induced occupational asthma. AB - BACKGROUND: Diisocyanates are among the most frequent causes of occupational asthma in industrialized countries. Early diagnosis of diisocyanate asthma followed by prompt termination of chemical exposure can prevent chronic morbidity due to persistent asthma. The accurate diagnosis of diisocyanate asthma requires a systematic approach that combines information obtained from the occupational history, immunologic tests and physiologic studies. METHODS: The advantages, limitations and validity of various methods and diagnostic guidelines utilized in the evaluation of diisocyanate asthma are reviewed. RESULTS: Recommended methods for evaluation of diisocyanates asthma are similar to approaches for other causative agents. Serologic assays of specific IgE are specific but insensitive diagnostic markers of diisocyanate asthma. If possible, workers should be evaluated, while at work, in order to demonstrate work-related changes in lung function associated with diisocyanate exposures. Specific bronchoprovocation challenge testing with diisocyanates, is reserved for situations where the diagnosis cannot be confirmed at work. Such tests can be performed safely but should be conducted exclusively at specialized centers by experienced personnel. CONCLUSIONS: Published diagnostic guidelines for occupational asthma are directly applicable to the evaluation of diisocyanate asthma. PMID- 10470012 TI - Possible discrepancies between dicentric chromosome frequencies and recorded ionizing radiation doses: in vivo study. AB - BACKGROUND: The available data on the effects of radiation on humans are limited to the reports of accidental exposure or studies of patients under diagnostic and therapeutic treatment; few reports refer to a occupationally exposed population groups. METHODS: The research was conducted on 66 subjects employed in the Department of Nuclear Medicine and 41 nonemployed controls. For each of them chromosomal analysis and gamma-spectrometer analysis of 24-hour urine were carried out. Exposure doses were measured using film-badge dosimetry. RESULTS: The comparison of dosimetric data obtained by film-badge measurements and the frequency of dicentric chromosomes in each subject revealed no correlation between the two observed parameters. CONCLUSIONS: In view of the above results, the subjects should be studied within job/task groups as they are far more likely to receive comparable doses. PMID- 10470013 TI - Blood lead levels among children of lead-exposed workers: A meta-analysis. AB - BACKGROUND: To further assess the utility of targeted blood lead screening for children from households with members having occupational lead exposures, we conducted a meta-analysis of all available reports of take-home lead exposures. Our objective was to estimate the blood lead levels among U.S. children (ages 1 5) from households with lead-exposed workers. METHODS: Reports considered for inclusion were cited in Medline, Toxline, Excerpta Medica, and Bio-Med plus all unpublished reports available at the National Institute for Occupational Safety and Health through 1994. The a priori criteria for inclusion of U.S. reports required their having data on: (1) venous blood lead levels for children, (2) children's ages, (3) data for at least five children, (4) workers' occupations, (5) workers' blood lead levels, and (6) data collection methods. RESULTS: Based on a meta-analysis of 10 reports from 1987 through 1994, the children (n=139) of lead-exposed workers (n=222) had a geometric mean blood lead level of 9.3 microg/dL compared to a U.S. population geometric mean of 3.6 microg/dL (P=0.0006). Also in this group, 52% of the children had blood lead levels (BLLs) >/= 10 microg/dL compared to 8.9% in the U.S. (P=.0010), and 21% of the children had BLLs >/= 20 microg/dL compared to 1.1% in the U.S. (P=. 0258). CONCLUSIONS: We estimate, based on 1981-83 survey data, that there are about 48,000 families with children under six living with household members occupationally exposed to lead. If the findings from this meta-analysis (admittedly limited by small numbers) are generalizable, about half of the young children in these families may have BLLs >/= 10 microg/dL. Data were too sparse to determine if children of workers with elevated blood leads were at greater risk than children whose parents were only known to be lead exposed. Our findings support the position that children of lead-exposed workers should be targeted for blood lead screening. Am. J. Ind. Med. 36:475-481, 1999. Published 1999 Wiley-Liss, Inc. PMID- 10470014 TI - Concerns and assumptions of labor and management in the dry-cleaning industry. PMID- 10470015 TI - Functional dissection of nebulette demonstrates actin binding of nebulin-like repeats and Z-line targeting of SH3 and linker domains. AB - Nebulette, a 107 kDa protein associated with the I-Z-I complex of cardiac myofibrils, may play an important role in the assembly of the Z-line. Determination of the complete primary structure of 1011 residue human fetal nebulette reveals a four-domain layout similar to skeletal muscle nebulin: a short N-terminal domain, followed by 22 nebulin-like repeats that are linked to a C-terminal Src homology 3 (SH3) domain via a short linker domain. To elucidate the mechanisms of assembly for nebulette in the Z-line, the complete coding sequence or fusions of nebulette domains with green fluorescent protein (GFP) were expressed in cardiomyocytes and fibroblasts. The complete protein localized to Z-lines in cardiac cells and to dense bodies in nonmuscle cells. The GFP repeat domain forms bundles that are associated with actin filaments in both cell types and disrupts the microfilament network. In contrast, the GFP-repeat plus linker shows limited interaction with dense bodies in nonmuscle cells and the Z lines of cardiomyocytes. Interestingly, the tagged linker or SH3 is diffusely distributed in nonmuscle cells, but localizes to the Z-lines in cardiomyocytes. Supporting the cellular localization work, recombinant nebulette fragments bind to actin, tropomyosin, and alpha-actinin in in vitro binding assays. These results suggest the repeat domain contains actin binding functions and that the linker domain may target this interaction to Z-lines and dense bodies. Our data also indicate that the linker and SH3 domains can distinguish between dense bodies and Z-lines, suggesting that the ligands for their interactions are specific to these muscular substructures. PMID- 10470016 TI - Diatom gliding is the result of an actin-myosin motility system. AB - Diatoms are a group of unicellular microalgae that are encased in a highly ornamented siliceous cell wall, or frustule. Pennate diatoms have bilateral symmetry and many genera possess an elongated slit in the frustule called the raphe, a feature synonymous with their ability to adhere and glide over a substratum, a process little understood. We have used cytoskeleton-disrupting drugs to investigate the roles of actin, myosin, and microtubules in diatom gliding or motility. No effect on diatom gliding was observed using the cytochalasins, known actin inhibitors, or the microtubule-inhibitors oryzalin and nocodazole. The latrunculins are a new group of anti-actin drugs, and we show here that they are potent inhibitors of diatom gliding, resulting in the complete disassociation of the raphe-associated actin cables. The recovery of actin staining and motility following latrunculin treatment was extremely fast. Cells exposed to latrunculin for 12 h recovered full function and actin staining within 5 sec of the drug being removed, demonstrating that the molecular components required for this motility system are immediately available. Butanedione monoxime (BDM), a known myosin inhibitor, also reversibly inhibited diatom gliding in a manner similar to the latrunculins. This work provides evidence that diatom gliding is based on an actin/myosin motility system. PMID- 10470017 TI - Microtubules in Xenopus oocytes are oriented with their minus-ends towards the cortex. AB - Despite lacking centrosomes, stage VI Xenopus oocytes contain extensive networks of cytoplasmic microtubules (MTs). To gain additional insight into the factors regulating MT organization during oogenesis, we have used electron microscopy and "hook decoration" to examine the distribution and orientation of MTs in Xenopus oocytes. A limited survey of two "undecorated" stage VI oocytes revealed 218 MTs in images covering approximately 2,500 microm(2), and indicated that the MT number density of the animal cytoplasm was greater than that of the vegetal cytoplasm. Examination of five "decorated" stage VI oocytes (three animal and five vegetal hemispheres) revealed 653 MTs. Of these, 76% could be scored as having exclusively counterclockwise (CCW) or clockwise (CW) hooks. In the animal hemispheres, 93% of the scored MTs exhibited CCW hooks when viewed from the direction of the cortex, indicating that most MTs were oriented with their minus ends out. MT orientation appeared relatively uniform throughout the animal cytoplasm: more than 90% of the scored MTs in the cortical (90%), subcortical (96%), or perinuclear (98%) cytoplasm were oriented with their minus-ends out. In the vegetal hemispheres, approximately 80% of the scored MTs exhibited CCW hooks, and thus were oriented with their minus-ends out; 96% of the scored MTs in stage III oocytes were oriented minus-end out. These observations support a model in which the cortex plays a significant role in MT nucleation and organization in Xenopus oocytes, and have significant implications for the MT-dependent transport and localization of cytoplasmic organelles and RNAs during oogenesis. PMID- 10470018 TI - Redistribution of surface-bound con A is quantitatively related to the movement of cells developing polarity. AB - Capping in cells developing polarity has been reinterpreted on the basis of a quantitative analysis of Concanavalin A (Con A) redistribution and cell movement in Walker carcinosarcoma cells. Several new features emerged. Based on the developing asymmetry in the distribution of surface-bound Con A, the direction of cell movement and the prospective front-tail polarity can already be predicted when the cell is spherical. Development of polarity by an initially spherical cell is associated with formation of two parts. The concentrically contracting part (prospective uropod) characterized by surface-associated Con A decreases in size, while the other part is cleared from Con A and grows into formerly unoccupied space. Surface-bound Con A shows isotropic centripetal movement towards the initial position of the centroid of the spherical cell rather than rearward movement. Therefore, the centroid of fluorescence intensity remains either stationary or moves marginally forward with respect to the initial position of the spherical cell. The amount and direction of cell movement measured correlates closely with values predicted by a theoretical model that assumes a unidirectional transfer of volume from a stationary contracting compartment into a protruding compartment. The results suggest that isotropic (cortical) contraction of the initially spherical cells and one-sided relaxation rather than unidirectional retrograde movement of ligand-receptor complexes produces movement in cells developing polarity. Reversible accumulation of surface-bound Con A at the uropod occurring to a similar extent in untreated and colchicine-treated cells is partly due to membrane folding and partly to movement in the plane of the membrane. PMID- 10470019 TI - Characterization of myosin-A and myosin-C: two class XIV unconventional myosins from Toxoplasma gondii. AB - Two class XIV unconventional myosins from Toxoplasma gondii, Myosin-A (TgM-A) and Myosin-C (TgM-C), were characterized in terms of their biochemical properties and their expression in quiescent and motile stages of the parasite life cycle. In cell fractionation studies, both myosins partitioned with the major organelle/cell membrane fraction, and extraction studies indicated that both were tightly associated with membrane domains as detergent was necessary for their solubilization. In addition, both TgM-A and TgM-C demonstrated a hallmark feature of myosins in their ability to bind actin in the absence but not the presence of ATP. In parasites residing within the host cell parasitophorous vacuole, TgM-A was detected by immunofluorescence microscopy as a bright spot near the apical pole of the parasite. This pattern underwent a subtle change as the parasites became motile, with TgM-A then localizing more intimately with the parasite cell membrane domain in apically disposed spots or patches, consistent with the role of this myosin in gliding motility. TgM-C showed a distinct localization to the juxtanuclear region towards the apical pole of the parasite, consistent with an association with the Golgi apparatus. PMID- 10470020 TI - C-terminal domain of the mitotic apparatus protein p62 targets the protein to the nucleolus during interphase. AB - Mitotic apparatuses from sea urchin embryos contain a protein (p62), previously shown to be required for mitotic progression. This protein localizes to the mitotic apparatus during cell division in urchin embryos and mammalian tissue culture cells. We show here by immunofluorescence that p62 is localized to the nucleus of mammalian cells during interphase and is highly concentrated in nucleoli. In addition, a fusion protein composed of full-length p62 and green fluorescent protein also localizes to nucleoli when expressed in COS-7 cells in culture. Analysis of the primary sequence of p62 reveals three distinct domains of the protein based on amino acid charge distribution: the acidic N-terminal domain, the basic C-terminal domain, and the central, M-domain, which contains alternating subdomains of clusters of acidic and basic residues. To identify the domain important for nucleolar localization during interphase, specific domains of p62 alone, or in combination with each other or with beta-galactosidase were fused to green fluorescent protein. Following confirmation of the fusion constructs by sequence analysis, the constructs were expressed in mammalian cells, expression was confirmed by immunoblotting, and the fusion proteins were localized via fluorescence microscopy. The data demonstrate that the C-terminal domain of p62 is both necessary and sufficient for the nuclear localization and nucleolar binding of p62 that is observed during interphase. PMID- 10470022 TI - Novel vaccine strategy demonstrates potential for Alzheimer's disease. PMID- 10470021 TI - Information overflow from discovery to development. PMID- 10470023 TI - Riboflavin binding proteins as chiral selectors in HPLC and CE. AB - The term riboflavin binding proteins (RfBPs) is applied to several molecular species that play the important role of vitamin delivery to the developing embryo, thus becoming essential for the survival of the fetus. In addition to this physiological significance, these proteins have recently been found to be successful chiral selectors. In this review, the authors address the use of such proteins, both as columns for high performance liquid chromatography (HPLC) and as additives in capillary electrophoresis (CE), for the enantioseparation of several racemic drugs. PMID- 10470024 TI - Taking polycation gene delivery systems from in vitro to in vivo. AB - It is widely believed that non-viral gene delivery systems may improve safety and overcome tissue-tropism limitations associated with viral-based gene therapies. Cationic liposome-based gene delivery currently accounts for 9-12% of ongoing gene therapy clinical trials in the United States and Europe. Polycation-based gene delivery is at an earlier development stage. However, both in vitro and in vivo studies have demonstrated that this is an area of much promise. Complexes of polycations with DNA result in major improvements in the control of size, charge, and the hydrophilic-lipophilic characteristics of the transfecting species, when compared with other non-viral systems. This review serves as an introduction to the current status of this field. PMID- 10470025 TI - Absorption prediction from physicochemical parameters. AB - Modern drug design not only focuses on the pharmacological activity of a compound but also considers its ability to be absorbed and to reach its site of action. The prediction of in vivo barrier permeabilities, particularly intestinal absorption and blood-brain barrier passage, are substantial concerns in the development of new drug compounds. For several decades, n-octanol-water partition coefficients dominated absorption prediction. In recent years, the basic physicochemical parameters describing both membrane permeability and lipophilicity have been established. This review provides an outline of some selected absorption-prediction models with emphasis on intestinal absorption. PMID- 10470027 TI - Automated knowledge refinement for rule-based formulation expert system. PMID- 10470026 TI - Monitor: progress and profiles. AB - Monitor provides an insight into the latest developments in pharmaceutical science and technology through brief synopses of recent presentations, publications and patents, and expert commentaries on the latest technologies. There are two sections: Progress summarizes the latest developments in pharmaceutical process technology, formulation, analytical technology, sterilization, controlled drug delivery systems and regulatory issues; Profiles offers expert commentary on emerging technologies, novel processes and strategic, organizational and logistic issues underlying pharmaceutical R&D. PMID- 10470028 TI - Protein misfolding, evolution and disease. PMID- 10470029 TI - Serpentine receptors and STAT activation: more than one way to twin a STAT. PMID- 10470030 TI - The TBP-like factor: an alternative transcription factor in metazoa? AB - Protein sequence analysis has revealed a family of TATA-binding-protein (TBP) like factors (TLFs) in metazoan organisms. Modelling of the three-dimensional structure of these TLFs suggests that they form an asymmetric saddle-like structure and that, unlike TBP, TLFs might bind to DNA sequences other than classical TATA boxes. Thus, the existence of TLFs presents a challenge to the doctrine that TBP is a universal regulator of transcription in metazoans. PMID- 10470031 TI - The GoLoco motif: a Galphai/o binding motif and potential guanine-nucleotide exchange factor. PMID- 10470032 TI - G-patch: a new conserved domain in eukaryotic RNA-processing proteins and type D retroviral polyproteins. PMID- 10470033 TI - Towards the three-dimensional structure of voltage-gated potassium channels. AB - Electrical excitability is a fundamental property of the neuromuscular systems of metazoans. The varied response of neurons to electrical excitation is largely accounted for by a diverse set of voltage-gated potassium (KV) channels in the excitable membrane. The complete structure of a KV channel is not yet available. However, recent structural biological experiments have begun to provide new insight into how specific KV channels are formed and regulated, and how they function and interact with other proteins. In particular, the selectivity of KV channels for K+ and suggestions as to how these structural elements might assemble into a functional KV channel are discussed. PMID- 10470034 TI - p21-activated protein kinase: a crucial component of morphological signaling? AB - The mechanisms by which Rho family GTPases (Rho, Rac and Cdc42) regulate coordinated changes to the actin cytoskeleton are being elucidated. This review will focus on the current evidence that the p21-activated kinases (PAKs) are involved in regulating some of the diverse cytoskeletal changes induced by Rac and Cdc42. PAKs have been shown to be required for processes including neurite formation and axonal guidance, development of cell polarity and motile responses. Signaling molecules interacting with PAKs that might contribute to the regulation of such processes have recently been identified. PMID- 10470035 TI - Tripeptidyl peptidases: enzymes that count. AB - Protein degradation is essential for the life and death of every cell. Proteins are broken down to their constitutive amino acids by a succession of peptidases, both in lysosomes and in the cytosol. Tripeptidyl-peptidases I and II are enzymes that can 'count to three' and release N-terminal tripeptides from oligopeptides generated by different endopeptidases. The tripeptides are then degraded by other exopeptidases to release amino acids and dipeptides. Mutations in tripeptidyl peptidase I have recently been associated with a lysosomal storage disease, late infantile neuronal ceroid lipofuscinosis. PMID- 10470036 TI - The enzymatic biotinylation of proteins: a post-translational modification of exceptional specificity. AB - Biotin is a coenzyme essential to all life forms. The vitamin has biological activity only when covalently attached to certain key metabolic enzymes. Most organisms have only one enzyme for attachment of biotin to other proteins and the sequences of these proteins and their substrate proteins are strongly conserved throughout nature. Structures of both the biotin ligase and the biotin carrier protein domain from Escherichia coli have been determined. These, together with mutational analyses of biotinylated proteins, are beginning to elucidate the exceptional specificity of this protein modification. PMID- 10470037 TI - Protein modelling for all. PMID- 10470039 TI - The tip of a molecular syringe: response PMID- 10470038 TI - The tip of a molecular syringe. PMID- 10470040 TI - Extracellular sensors and inducible protective mechanisms. PMID- 10470041 TI - Defence mechanisms against DNA-damaging agents in Escherichia coli. PMID- 10470042 TI - The pathogenic potential of endogenous retroviruses: facts and fantasies. AB - Endogenous retroviruses are descendants of viruses that became cellular genes by integration into their host's genome. They still contribute to pathogenicity as a partner in recombination events, by de novo insertion after mobilization followed by activation of downstream proto-oncogenes, or by gene disruption. Re-expression of viral proteins accompanied by loss of immune tolerance could induce immune disturbances. PMID- 10470043 TI - The interaction between RTX toxins and target cells. AB - RTX toxins are important virulence factors produced by a wide range of Gram negative bacteria. They fall into two categories: the hemolysins, which affect a variety of cell types, and the leukotoxins, which are cell-type- and species specific. These toxins offer interesting models for targeting, insertion and translocation of aqueous proteins into lipid membranes. PMID- 10470044 TI - Why is Deinococcus radiodurans so resistant to ionizing radiation? AB - When exponential-phase cultures of Deinococcus radiodurans are exposed to a 5000 Gray dose of gamma radiation, individual cells suffer massive DNA damage. Despite this insult to their genetic integrity, these cells survive without loss of viability or evidence of mutation, repairing the damage by as-yet-poorly understood mechanisms. PMID- 10470045 TI - Defining the requirements for immunological control of mycobacterial infections. AB - Tuberculosis remains a major public health problem throughout the world. Research into the immunological basis of the host-pathogen relationship has recently benefited from the fascinating convergence of genetic data from mouse models and from humans. Latency - the seemingly quiescent phase of bacterial persistence - is the central problem in controlling tuberculosis and will be the next frontier of research. PMID- 10470046 TI - Perspective: human contact patterns and the spread of airborne infectious diseases. AB - Networks of social contacts channel the transmission of airborne infections. Emerging insights from fields of science as diverse as mathematics, population biology and the social sciences are beginning to reveal how the contact pattern of the hosts determines the spread and evolution of airborne infectious agents. PMID- 10470047 TI - The influence of dsRNA viruses on the biology of plant pathogenic fungi. AB - Double-stranded RNA viruses are ubiquitous in fungi. They are non-infective and, like most prokaryotic plasmids, are only transmitted to compatible strains via cell fusion. Most are cryptic, but some with an established phenotype, such as the hypoviruses of the chestnut-blight fungus, have been studied for their potential as biological control agents of fungi. PMID- 10470048 TI - Cause and consequence of becoming persistentPersistent viral infections edited by R. Ahmed and I.S.Y. chen PMID- 10470050 TI - Uganda gives lessons in stemming the AIDS epidemic. PMID- 10470051 TI - No increase in demyelinating diseases after hepatitis B vaccination. PMID- 10470052 TI - The dangers of extended PCR in the clinic. PMID- 10470053 TI - Lack of apoptosis in mice with ALS. PMID- 10470054 TI - Italian changes are not surprising. PMID- 10470055 TI - Grants will determine European malaria vaccine future. PMID- 10470056 TI - India fast-tracks vaccine candidates. PMID- 10470057 TI - German cutbacks hurt science. PMID- 10470059 TI - Universities offer animal rights courses. PMID- 10470058 TI - Advocates push for new NIH office. PMID- 10470060 TI - New rules for lazy professors. PMID- 10470061 TI - Human therapeutic cloning. AB - Somatic cell nuclear 'reprogramming' in livestock species is now routine in many laboratories. Here, Robert Lanza, Jose Cibelli and Michael West discuss how these techniques may soon be used to clone genetically matched cells and tissues for transplantation into patients suffering from a wide range of disorders that result from tissue loss or dysfunction. PMID- 10470062 TI - Dendritic Cells: Biology and Clinical Applications. PMID- 10470063 TI - The Sceptical Witness. PMID- 10470064 TI - Feeling sleepy? It's in your genes! PMID- 10470065 TI - Does our DNA determine when we sleep? AB - The discovery and study of three kindreds with advanced sleep phase disorder shed light on how we can inherit tendencies to be early morning or late night kinds of people (pages 1062-1065). PMID- 10470066 TI - From Agouti to Pomc--100 years of fat blonde mice. PMID- 10470067 TI - HIV-1 Nef protein: An invitation to a kill. PMID- 10470068 TI - Nucleotides and cd39: principal modulatory players in hemostasis and thrombosis. PMID- 10470070 TI - Another tetracycline - induced transformation. PMID- 10470069 TI - Turning on tumors to study cancer progression. PMID- 10470071 TI - Induction of an oncogenic fusion protein by a viral gene--a new chapter in an old story. PMID- 10470072 TI - Spotting a killer's target in autoimmune diabetes. PMID- 10470073 TI - Cocaine addiction therapy--are we partially there? PMID- 10470074 TI - In stroke, complement will get you nowhere. PMID- 10470075 TI - HIV-1 Nef mediates lymphocyte chemotaxis and activation by infected macrophages. AB - Infection of macrophage lineage cells is a feature of primate lentivirus replication, and several properties of primate lentiviruses seem to have evolved to promote the infection of macrophages. Here we demonstrate that the accessory gene product Nef induces the production of two CC-chemokines, macrophage inflammatory proteins 1alpha and 1beta, by HIV-1-infected macrophages. Adenovirus mediated expression of Nef in primary macrophages was sufficient for chemokine induction. Supernatants from Nef-expressing macrophages induced both the chemotaxis and activation of resting T lymphocytes, permitting productive HIV-1 infection. These results indicate a role for Nef in lymphocyte recruitment and activation at sites of virus replication. PMID- 10470076 TI - Allo-immunization elicits CD8+ T cell-derived chemokines, HIV suppressor factors and resistance to HIV infection in women. AB - We assessed the potential for an allogeneic-based vaccine against HIV infection in women who were allo-immunized with their partners' mononuclear leucocytes to prevent spontaneous recurrent abortion. Within 1 month of allo-immunization, there was significant upregulation in the concentrations of CD8 cell-derived suppressor factor activity, RANTES, and macrophage inflammatory proteins 1alpha and 1beta. Allo-immunization also downregulated the proportion of cells with CCR5 and CXCR4 receptors. We also found a dose-dependent decrease in HIV infectivity of CD4+ cells in vitro after allo-immunization with both primary and T-cell line adapted HIV-1. This study provides a rational basis for an alternative or complementary strategy of allo-immunization against HIV infection. PMID- 10470077 TI - Targeted disruption of cd39/ATP diphosphohydrolase results in disordered hemostasis and thromboregulation. AB - CD39, or vascular adenosine triphosphate diphosphohydrolase, has been considered an important inhibitor of platelet activation. Unexpectedly, cd39-deficient mice had prolonged bleeding times with minimally perturbed coagulation parameters. Platelet interactions with injured mesenteric vasculature were considerably reduced in vivo and purified mutant platelets failed to aggregate to standard agonists in vitro. This platelet hypofunction was reversible and associated with purinergic type P2Y1 receptor desensitization. In keeping with deficient vascular protective mechanisms, fibrin deposition was found at multiple organ sites in cd39-deficient mice and in transplanted cardiac grafts. Our data indicate a dual role for adenosine triphosphate diphosphohydrolase in modulating hemostasis and thrombotic reactions. PMID- 10470078 TI - Uteroglobin is essential in preventing immunoglobulin A nephropathy in mice. AB - The molecular mechanism(s) of immunoglobulin A (IgA) nephropathy, the most common primary renal glomerular disease worldwide, is unknown. Its pathologic features include hematuria, high levels of circulating IgA-fibronectin (Fn) complexes, and glomerular deposition of IgA, complement C3, Fn and collagen. We report here that two independent mouse models (gene knockout and antisense transgenic), both manifesting deficiency of an anti-inflammatory protein, uteroglobin (UG), develop almost all of the pathologic features of human IgA nephropathy. We further demonstrate that Fn-UG heteromerization, reported to prevent abnormal glomerular deposition of Fn and collagen, also abrogates both the formation of IgA-Fn complexes and their binding to glomerular cells. Moreover, UG prevents glomerular accumulation of exogenous IgA in UG-null mice. These results define an essential role for UG in preventing mouse IgA nephropathy and warrant further studies to determine if a similar mechanism(s) underlies the human disease. PMID- 10470079 TI - Identification of an MHC class I-restricted autoantigen in type 1 diabetes by screening an organ-specific cDNA library. AB - Type 1 diabetes is an autoimmune disease in which the insulin-producing pancreatic beta cells are destroyed at an early age by an immune process that involves both CD4 and CD8 T lymphocytes. The identification of autoantigens in diabetes is very important for the design of antigen-specific immunotherapy. By screening a pancreatic islet cDNA library, we have identified the autoantigen recognized by highly pathogenic CD8 T cells in the non-obese diabetic mouse, one of the best animal models for human diabetes. This is the first identification, to our knowledge, of a CD8 T-cell epitope in an autoimmune disease. The peptide recognized by the cells is in the same region of the insulin B chain as the epitope recognized by previously isolated pathogenic CD4 T cells. This has very important implications for the potential use of insulin in preventative therapy. PMID- 10470080 TI - Anti-cancer activity of targeted pro-apoptotic peptides. AB - We have designed short peptides composed of two functional domains, one a tumor blood vessel 'homing' motif and the other a programmed cell death-inducing sequence, and synthesized them by simple peptide chemistry. The 'homing' domain was designed to guide the peptide to targeted cells and allow its internalization. The pro-apoptotic domain was designed to be nontoxic outside cells, but toxic when internalized into targeted cells by the disruption of mitochondrial membranes. Although our prototypes contain only 21 and 26 residues, they were selectively toxic to angiogenic endothelial cells and showed anti cancer activity in mice. This approach may yield new therapeutic agents. PMID- 10470081 TI - Cytomegalovirus US2 destroys two components of the MHC class II pathway, preventing recognition by CD4+ T cells. AB - Human cytomegalovirus (HCMV) is a ubiquitous herpesvirus that causes life threatening disease in patients who are immunosuppressed for bone marrow or tissue transplantation or who have AIDS (ref. 1). HCMV establishes lifelong latent infections and, after periodic reactivation from latency, uses a panel of immune evasion proteins to survive and replicate in the face of robust, fully primed host immunity. Monocyte/macrophages are important host cells for HCMV, serving as a latent reservoir and as a means of dissemination throughout the body. Macrophages and other HCMV-permissive cells, such as endothelial and glial cells, can express MHC class II proteins and present antigens to CD4+ T lymphocytes. Here, we show that the HCMV protein US2 causes degradation of two essential proteins in the MHC class II antigen presentation pathway: HLA-DR-alpha and DM-alpha. This was unexpected, as US2 has been shown to cause degradation of MHC class I (refs. 5,6), which has only limited homology with class II proteins. Expression of US2 in cells reduced or abolished their ability to present antigen to CD4+ T lymphocytes. Thus, US2 may allow HCMV-infected macrophages to remain relatively 'invisible' to CD4+ T cells, a property that would be important after virus reactivation. PMID- 10470082 TI - Catalytic activity of antibodies against factor VIII in patients with hemophilia A. AB - Hemophilia A is an X chromosome-linked recessive disorder resulting in defective or deficient factor VIII (FVIII) molecules, which, in its severe form, is a life threatening and crippling hemorrhagic disease. Infusion of homologous FVIII to patients with severe hemophilia A results, in 25% of patients, in the emergence of alloantibodies against FVIII (inhibitors)( ref. 1) that inhibit FVIII procoagulant activity by steric hindrance of the interaction of FVIII either with stabilizing molecules, with molecules essential for its activity or with activating molecules. Here, we report on the proteolysis of FVIII by alloantibodies of two patients with severe hemophilia A, demonstrating a previously unknown mechanism by which FVIII inhibitors may prevent the pro coagulant function of FVIII. The kinetic parameters of FVIII hydrolysis indicate a functional role for the catalytic immune response in the inactivation of FVIII in vivo. The characterization of alloantibodies against FVIII as site-specific proteases may provide new approaches to the treatment of FVIII inhibitors. PMID- 10470083 TI - MRP4: A previously unidentified factor in resistance to nucleoside-based antiviral drugs. AB - Dideoxynucleosides, which are potent inhibitors of HIV reverse transcriptase and other viral DNA polymerases, are a common component of highly active anti retroviral therapy (HAART) (ref. 1). Six reverse transcriptase inhibitors have been approved for human use: azidothymidine; 2'3'-dideoxycytidine; 2'3' dideoxyinosine; 2', 3'-didehydro-3'deoxythymidine; 2',3'-dideoxy-3'-thiacytidine; and 4-[2-amino-6-(cyclopropylamino)-9H-purin-9-yl]-2-cyclopentene-1-++ +metha nol. Although drug-resistant HIV strains resulting from genetic mutation have emerged in patients treated with HAART (ref. 1), some patients show signs of drug resistance in the absence of drug-resistant viruses. In our study of alternative or additional mechanisms of resistance operating during antiviral therapy, overexpression and amplification of the MRP4 gene correlated with ATP-dependent efflux of PMEA (9-(2-phosphonylmethoxyethyl)adenine) and azidothymidine monophosphate from cells and, thus, with resistance to these drugs. Overexpression of MRP4 mRNA and MRP4 protein severely impaired the antiviral efficacy of PMEA, azidothymidine and other nucleoside analogs. Increased resistance to PMEA and amplification of the MRP4 gene correlated with enhanced drug efflux; transfer of chromosome 13 containing the amplified MRP4 gene conferred resistance to PMEA. MRP4 is the first transporter, to our knowledge, directly linked to the efflux of nucleoside monophosphate analogs from mammalian cells. PMID- 10470084 TI - Genetic capsid modifications allow efficient re-targeting of adeno-associated virus type 2. AB - The human parvovirus adeno-associated virus type 2 (AAV2) has many features that make it attractive as a vector for gene therapy. However, the broad host range of AAV2 might represent a limitation for some applications in vivo, because recombinant AAV vector (rAAV)-mediated gene transfer would not be specific for the tissue of interest. This host range is determined by the binding of the AAV2 capsid to specific cellular receptors and/or co-receptors. The tropism of AAV2 might be changed by genetically introducing a ligand peptide into the viral capsid, thereby redirecting the binding of AAV2 to other cellular receptors. We generated six AAV2 capsid mutants by inserting a 14-amino-acid targeting peptide, L14, into six different putative loops of the AAV2 capsid protein identified by comparison with the known three-dimensional structure of canine parvovirus. All mutants were efficiently packaged. Three mutants expressed L14 on the capsid surface, and one efficiently infected wild-type AAV2-resistant cell lines that expressed the integrin receptor recognized by L14. The results demonstrate that the AAV2 capsid tolerates the insertion of a nonviral ligand sequence. This might open new perspectives for the design of targeted AAV2 vectors for human somatic gene therapy. PMID- 10470085 TI - Painful stimulation suppresses joint inflammation by inducing shedding of L selectin from neutrophils. AB - Although the inflammatory response is essential for protecting tissues from injury and infection, unrestrained inflammation can cause chronic inflammatory diseases such as arthritis, colitis and asthma. Physiological mechanisms that downregulate inflammation are poorly understood. Potent control might be achieved by regulating early stages in the inflammatory response, such as accumulation of neutrophils at the site of injury, where these cells release chemical mediators that promote inflammatory processes including plasma extravasation, bacteriocide and proteolysis. To access an inflammatory site, neutrophils must first adhere to the vascular endothelium in a process mediated in part by the leukocyte adhesion molecule L-selectin. This adhesion is prevented when L-selectin is shed from the neutrophil membrane. Although shedding of L-selectin is recognized as a potentially important mechanism for regulating neutrophils, its physiological function has not been demonstrated. Shedding of L-selectin may mediate endogenous downregulation of inflammation by limiting neutrophil accumulation at inflammatory sites. Here we show that activation of nociceptive neurons induces shedding of L-selectin from circulating neutrophils in vivo and that this shedding suppresses an ongoing inflammatory response by inhibiting neutrophil accumulation. These findings indicate a previously unknown mechanism for endogenous feedback control of inflammation. Failure of this mechanism could contribute to the etiology of chronic inflammatory disease. PMID- 10470086 TI - Familial advanced sleep-phase syndrome: A short-period circadian rhythm variant in humans. AB - Biological circadian clocks oscillate with an approximately 24-hour period, are ubiquitous, and presumably confer a selective advantage by anticipating the transitions between day and night. The circadian rhythms of sleep, melatonin secretion and body core temperature are thought to be generated by the suprachiasmatic nucleus of the hypothalamus, the anatomic locus of the mammalian circadian clock. Autosomal semi-dominant mutations in rodents with fast or slow biological clocks (that is, short or long endogenous period lengths; tau) are associated with phase-advanced or delayed sleep-wake rhythms, respectively. These models predict the existence of familial human circadian rhythm variants but none of the human circadian rhythm disorders are known to have a familial tendency. Although a slight 'morning lark' tendency is common, individuals with a large and disabling sleep phase-advance are rare. This disorder, advanced sleep-phase syndrome, is characterized by very early sleep onset and offset; only two cases are reported in young adults. Here we describe three kindreds with a profound phase advance of the sleep-wake, melatonin and temperature rhythms associated with a very short tau. The trait segregates as an autosomal dominant with high penetrance. These kindreds represent a well-characterized familial circadian rhythm variant in humans and provide a unique opportunity for genetic analysis of human circadian physiology. PMID- 10470087 TI - Obesity in the mouse model of pro-opiomelanocortin deficiency responds to peripheral melanocortin. AB - Pro-opiomelanocortin (POMC)-derived peptides (the melanocortins adrenocorticotropin, alpha-, beta- and gamma-melanocyte stimulating hormone; and the endogenous opioid beta-endorphin) have a diverse array of biological activities, including roles in pigmentation, adrenocortical function and regulation of energy stores, and in the immune system and the central and peripheral nervous systems. We show here that mice lacking the POMC-derived peptides have obesity, defective adrenal development and altered pigmentation. This phenotype is similar to that of the recently identified human POMC-deficient patients. When treated with a stable alpha-melanocyte-stimulating hormone agonist, mutant mice lost more than 40% of their excess weight after 2 weeks. Our results identify the POMC-null mutant mouse as a model for studying the human POMC-null syndrome, and indicate the therapeutic use of peripheral melanocortin in the treatment of obesity. PMID- 10470088 TI - The type of somatic mutation at APC in familial adenomatous polyposis is determined by the site of the germline mutation: a new facet to Knudson's 'two hit' hypothesis. AB - APC is often cited as a prime example of a tumor suppressor gene. Truncating germline and somatic mutations (or, infrequently, allelic loss) occur in tumors in FAP (familial adenomatous polyposis). Most sporadic colorectal cancers also have two APC mutations. Clues from attenuated polyposis, missense germline variants with mild disease and the somatic mutation cluster region (codons 1,250 1,450) indicate, however, that APC mutations might not result in simple loss of protein function. We have found that FAP patients with germline APC mutations within a small region (codons 1,194-1,392 at most) mainly show allelic loss in their colorectal adenomas, in contrast to other FAP patients, whose 'second hits' tend to occur by truncating mutations in the mutation cluster region. Our results indicate that different APC mutations provide cells with different selective advantages, with mutations close to codon 1,300 providing the greatest advantage. Allelic loss is selected strongly in cells with one mutation near codon 1,300. A different germline-somatic APC mutation association exists in FAP desmoids. APC is not, therefore, a classical tumor suppressor. Our findings also indicate a new mechanism for disease severity: if a broader spectrum of mutations is selected in tumors, the somatic mutation rate is effectively higher and more tumors grow. PMID- 10470091 TI - ON THE MARKET. PMID- 10470089 TI - An association between viral genes and human oncogenic alterations: the adenovirus E1A induces the Ewing tumor fusion transcript EWS-FLI1. AB - Malignant transformation of human cells requires the accumulation of multiple genetic alterations, such as the activation of oncogenes and loss of function of tumor suppressor genes or those related to genomic instability. Among the genetic alterations most frequently found in human tumors are chromosomal translocations that may result in the expression of chimeric products with transforming capability or are able to change the expression of oncogenes. We show here that the adenovirus early region 1A (E1A) gene can induce a specific human fusion transcript (EWS-FLI1) that is characteristic of Ewing tumors. This fusion transcript was detected by RT-PCR in normal human fibroblasts and keratinocytes after expression of the adenovirus E1A gene, as well as in human cell lines immortalized by adenoviruses. Cloning and sequencing of the RT-PCR product showed fusion points between EWS and FLI1 cDNA identical to those detected in Ewing tumors. In addition, we detected a chimeric protein by western blot analysis and immunoprecipitation and a t(11,22) by fluorescent in situ hybridization. This association between a single viral gene and a specific human fusion transcript indicates a direct link between viral genes and chromosome translocations, one of the hallmarks of many human tumors. PMID- 10470090 TI - Equivalent amplification of intrinsically variable nucleic acid sequences by multiple-primer-induced overlapping amplification assay: applications for universal detection and quantitation. PMID- 10470093 TI - Inhibition of ribonucleotide reductase by a new class of isoindole derivatives: drug synergism with cytarabine (Ara-C) and induction of cellular apoptosis. AB - The hydroxyisoindole dione derivatives ISID and MISID are new compounds with structures resembling purines and possessing a hydroxamic acid moiety which is the pharmacophore of hydroxyurea (HU), an inhibitor of ribonucleotide reductase (RR). ISID and MISID exhibited 100- to 500-fold higher cytotoxicity as compared to HU against cell lines sensitive (CEM/0) or resistant to ara-C (CEM/ara-C/7A; CEM/dCk[-]). Both MISID and ISID showed significant inhibitory activity of ribonucleotide reductase (RR). Treatment of CEM/0 cells with 10 microM ISID showed a linear decrease in all the dNTPs leading to a complete depletion by 4 hours with no recovery of enzymatic activity of RR up to 48 hours in the presence of the drug, suggesting an irreversible inhibition of this enzyme. However, 10 microM MISID caused a significant time dependent, but reversible inhibition of RR in a whole cell assay in CEM/0 cells. Pretreatment of CEM/0 cells with 10 microM MISID for 1 hour increased cellular ara-CTP concentrations approximately 2-fold as compared to untreated controls. However, a reduction in intracellular ara-CTP concentration was observed following a commensurate depletion of ATP in these cells after 4 hrs of ISID pretreatment. Similarly, the ara-CTP concentration was augmented by 1.6-fold following pretreatment of CEM/0 cells with 10 microM MISID for 4 hours. Significant apoptotic cell death was detected in CEM/0 cells treated with ara-C, ISID or MISID alone or in combination. Ara-C treatment induced HMW (high molecular weight) DNA fragmentation at earlier times which subsequently led to oligonucleosomal DNA fragmentation by 48 hrs. The sequential treatment of CEM/0 cells with MISID followed by ara-C resulted in increased DNA fragmentation in the 2.0 to 4.0 Kb range in comparison to those cells treated with either ara-C or MISID alone. The increased apoptotic cell death explained the synergistic cytotoxicity of the combination of ara-C and MISID against CEM/0 cells observed earlier. We conclude that the inhibition of RR by these agents induces leukemic cell apoptosis, a mechanism which is further potentiated when these RR inhibitors are combined with ara-C. Since new compounds do not require activation, as do other clinically useful RR inhibitors, further studies for their potential use against leukemias and solid tumors are warranted. PMID- 10470095 TI - Proto-oncogene N-myc promoter is down regulated by the Wilms' tumor suppressor gene WT1. AB - The Wilms' tumor 1 (WT1) gene is a tumor suppressor gene that encodes a zinc finger transcription factor. WT1 represses transcription of several growth factors and growth factor receptors. The N-myc proto-oncogene encodes a transcription factor which regulates cell growth and differentiation. N-myc is coexpressed with WT1 in the developing kidney and is overexpressed in many Wilms' tumors. Here, we show that the proto-oncogene N-myc promoter was down-regulated by WT1 in transient transfection assays. However, mutant WT1 (R394W) which has a mutation in the DNA binding domain could not repress the N-myc promoter. Electrophoretic mobility shift assays showed that the oligonucleotides containing the WT1 motifs could bind recombinant WT1 proteins. This suggests that the repression of the N-myc promoter is mediated through the WT1 binding sites. This finding may help to elucidate the relationship of WT1 and N-myc in tumorigenesis and renal development. PMID- 10470094 TI - Hepatocyte growth factor (HGF) and receptor (c-met) in normal and malignant astrocytic cells. AB - BACKGROUND: Hepatocyte growth factor (HGF) is a multifunctional peptide that binds to a specific receptor, c-met. Both HGF and c-met have been identified in normal brain and on glial tumors. The purpose of this study is to further define the biologic importance of HGF and c-met on normal and malignant glial cells grown in vitro. MATERIALS AND METHODS: Nine human malignant glioma-derived tumor cell cultures and cultures of astrocytes derived from normal brain were examined for c-met and HGF transcripts using Northern blot or RT-PCR analysis. Cellular invasiveness was quantitated by mechanical assay and mitogenesis was determined by cell count. RESULTS: C-met was expressed in five of seven malignant glioma derived tumor cell cultures and in both normal astrocyte cultures. HGF transcript was not detected in any of the cell cultures. HGF supplementation enhanced invasiveness in c-met positive cell lines and did not alter cellular mitogenesis in the assayed cultures. CONCLUSIONS: These findings suggest that HGF is a potent stimulator of invasiveness in c-met positive malignant glioma-derived tumor cells and is not an active cytokine with regards to in vitro glial cell proliferation. HGF may therefore stimulate glioma cellular invasion in vivo through binding to its receptor and by activating tyrosine kinase secondary messengers. PMID- 10470096 TI - Growth of human lung adenocarcinoma in nude mice is influenced by various types of dietary fat and vitamin E. AB - Studies have shown effects of dietary lipids on carcinogenesis and tumour progression. Different mechanisms for the inhibitory effect of n-3 fatty acids (FA) have been proposed. The inhibition of the growth of subcutaneously transplanted A427 lung adenocarcinoma cells in athymic nude mice may occur due to an increased level of lipid peroxidation products and is the object of this study. The nude mice were fed diets supplemented with corn oil (CO), olive oil (OO) or K85, a mixture of ethyl esters of n-3 FAs, mainly eicosapentaenoic acid (EPA, 20:5, n-3) and docosahexaenoic acid (DHA, 22:6, n-3). Tumours of the n-3 FA group showed reduced growth. Peroxidation products measured by the thiobarbituric acid reactive substances (TBARS) test showed higher levels in tumours from n-3 FA fed mice than in the other diet groups. The growth inhibitory effects and the elevated level of TBARS in the n-3 FA diet group were counteracted by vitamin E supplement in the diet. Cu/Zn-superoxide dismutase (SOD) activity in liver did not differ greatly among the diet groups. The Ki-67 labelling index (LI), indicating cell proliferation rate was significantly lower in the K85 diet group compared to the other diet groups. PMID- 10470097 TI - Tamoxifen and genistein synergistically down-regulate signal transduction and proliferation in estrogen receptor-negative human breast carcinoma MDA-MB-435 cells. AB - PURPOSE: Tamoxifen and genistein were tested for synergism in estrogen receptor- negative human breast carcinoma MDA-MB-435 cells because the two compounds decrease signal transduction activity through different biochemical mechanisms and arrest the cell cycle at different phases. MATERIALS AND METHODS: The combination effect of tamoxifen and genistein on signal transduction was determined by measuring IP3 concentrations and on cell proliferation and colony formation by growth inhibition assay and clonogenic assay. RESULTS: In growth inhibition assays, for tamoxifen and genistein in the carcinoma cells the IC50s were (mean +/- SE) 17 +/- 0.9 and 27 +/- 1.6 microM; in clonogenic assays the LC50s were 0.9 +/- 0.4 and 12.5 +/- 1.1 microM, respectively. When tamoxifen and genistein were simultaneously added to the cells, synergism was observed in growth inhibition, in cytotoxicity and in the reduction of inositol 1,4,5 trisphosphate concentration. CONCLUSION: The synergistic down-regulation of signal transduction by tamoxifen and genistein may explain, in part at least, the synergistic antiproliferative and cytotoxic actions of the two compounds. The synergism of tamoxifen and genistein may be of interest in the clinical treatment of breast carcinoma. PMID- 10470098 TI - Enhanced anti-metastatic efficacy of IL-2 activated NK (A-NK) cells with novel benzothiazoles. AB - We have previously shown that A-NK cells when locoregionally administered accumulate within established cancer metastases and establish direct contact with both tumor cells and microvascular endothelial cells. Nevertheless, the accumulation of adoptively transferred A-NK cells into established cancer metastases is not sufficient for therapeutic efficacy in the B16 melanoma model. We have therefore attempted to enhance the anti-metastatic therapeutic efficacy of adoptively transferred A-NK cells with standard anticancer chemotherapeutic agents. We have found that chemoimmunotherapy with A-NK cells plus cyclophosphamide to be more effective than A-NK cell adoptive immunotherapy alone. We have now built on these findings, by examining the ability of novel biologic response modifiers (low molecular weight benzothiazole compounds) to augment adoptive immunotherapy with A-NK cells. Two compounds KB-R4107 (4-methoxy 2-(4-t-butylphenyl)benzothiazole) and KB-R4250 (4-methoxy-2-(4 trifluoromethylphenyl)benzothiazole) enhanced reduction of B16 melanoma pulmonary metastases mediated by A-NK cell adoptive immunotherapy. Both compounds were administered for 5 days prior to administration of A-NK cells at 100 mg/kg p.o. All experimental groups initially contained at least 7 animals and were examined for tumor burden on day 10. With B16 melanoma cells administered on day 0 and A NK cells administered on Day 4, KB-R4107 and KB-R4250 yielded on average a 64% and 52% reduction in metastatic burden, respectively compared to an average 17% reduction using A-NK cells alone. In contrast these compounds did not diminish metastatic burden when administered alone. KB-R4107 and KB-R4250 are therefore low molecular weight, heterocyclic, biological response modifiers which can augment the anti-metastatic therapeutic effect of adoptively transferred A-NK cells. PMID- 10470099 TI - Inverse correlation between apoptotic (Fas ligand, caspase-3) and angiogenic factors (VEGF, microvessel density) in squamous cell lung carcinomas. AB - In order to explore whether apoptosis is associated with angiogenesis in lung cancer, immunohistochemistry was employed to determine the pro-apoptotic factors Fas ligand (FasL) and caspase-3 (Cas-3) in 70 squamous cell lung carcinomas. Furthermore, the vascular endothelial growth factor (VEGF) and the microvessel density (MVD) were analyzed. The comparison between MVD and the pro-apoptotic factors demonstrated that the apoptotic factors are inversely related to MVD (Cas 3: p = 0.011, FasL: not significant). In order to confirm this result, FasL and Cas-3 were also compared with the expression of VEGF. Again, an inverse correlation between VEGF and the pro-apoptotic factors was found (Cas-3: p = 0.019, FasL: p = 0.008). The inverse correlation between angiogenesis and apoptosis may be explained by the activation of pro-apoptotic and anti-angiogenic factors caused by hypoxia. PMID- 10470101 TI - Cell and tissue-type specific expression of Ras-related GTPase RhoB. AB - Ras-homologous (Rho) GTPases are involved in the regulation of a variety of cellular processes such as the organization of the actin cytoskeleton, malignant transformation and genotoxic stress-induced signaling. Here we show that, among the family of Rho GTPases, specifically rhoB mRNA expression is rapidly induced upon UV-irradiation, whereas the level of rac 1 and cdc42 mRNA is not affected. Increase in rhoB mRNA was accompanied by a approximately 4-fold increase in the amount of membrane-bound RhoB protein. Basal expression of rhoB mRNA appears to be cell-type specific with low amounts in rodent NIH 3T3, V79, H4IIE, CHO and human HaCat cells and comparably high amounts in monkey COS, human HeLa and HepG2 cells. In rabbit tissues, exceptionally high levels of rhoB mRNA and RhoB protein were found in lung whereas its expression was quite low in heart, liver, spleen and kidney. Variations in rhoB mRNA expression level are not due to cell-type specific differences in rhoB mRNA stability as shown by inhibitor experiments. However, transiently transfected rhoB promoter CAT construct was expressed at significantly higher level in HeLa and HepG2 as compared to NIH 3T3 and CHO cells. Thus, cell-type specific differences in the level of rhoB mRNA are likely to be due to variations in the transcriptional activity of the rhoB gene. The data indicate that, among the family of Rho GTPases, only the expression of rhoB is rapidly stimulated by genotoxic stress. Furthermore basal rhoB expression appears to be regulated in a cell and tissue-type specific manner. This may be related to yet unknown tissue-specific physiological function of RhoB. PMID- 10470100 TI - Anti-estrogenic activities of indole-3-carbinol in cervical cells: implication for prevention of cervical cancer. AB - BACKGROUND: Cervical cancer constitutes the second most common cancer in women. Estrogen promotes development of cervical cancer in cells infected with high risk human papillomaviruses (HPVs). We asked whether the phytochemical indole-3 carbinol (I3C) has anti-estrogenic activities in cervical cells with the goal of preventing cancer in HPV infected cells. MATERIALS AND METHODS: Using the cervical cancer cell line CaSki, we evaluated expression of HPV and cytochrome p450 (CYP) enzymes by Northern, RNase protection or quantitative RT-PCR. I3C binding to estrogen receptor was measured by competition with estradiol. Estrogen metabolites were measured by gas chromarography-mass spectrometry (GC-MS). RESULTS: Estradiol increased expression of HPV oncogenes whereas I3C and the estrogen metabolite 2-hydroxyestrone (2-OHE) abrogated the estrogen-increased expression of HPV oncogenes. Both I3C and 2-OHE competed with estradiol for estrogen receptor binding. I3C enhanced gene expression of CYP enzymes responsible for 2-hydroxylation of estrogen, and induced the formation of 2-OHE. CONCLUSION: I3C has anti-estrogenic activities which should prevent cancer in cervical cells. PMID- 10470102 TI - 20-Cyclopropyl-cholecalciferol vitamin D3 analogs: a unique class of potent inhibitors of proliferation of human prostate, breast and myeloid leukemia cell lines. AB - We have synthesized and studied the ability of a series of nine novel 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] analogs to inhibit clonal growth of myeloid leukemic cells (HL,60), prostate (LNCaP, PC-3 and DU-145) and breast (MCF-7) cancers cells. DU-145 cells were actively resistant to compounds (cmpd) with all of these modifications, but when we removed C-19 (E, 1,25-Dihydroxy-23E-ene-26,27 hexafluoro-19-nor-20-cyclopropy l- cholecalciferol) an analog resulted that was inhibitory against all three prostate cell lines, breast and HL-60 cell lines. Further analysis showed that pulse exposure (3 days, 10(-7) M) to this analog was enough to inhibit clonal growth of PC-3 cell by 50%. Furthermore, cmpd E increased the number of PC-3 cells in G1 and decreased the number in S phase. 1,25(OH)2D3 mediates its biological activities through specific binding to the vitamin D3 receptor (VDR) and subsequent association with vitamin D3 response elements (VDRE) in genes modulated by 1,25(OH)2D3. Several novel vitamin D3 cmpds have recently been identified which have 5- to 1000-fold greater abilities to induce differentiation and to inhibit proliferation of prostate cancer, breast cancer and HL-60 leukemic blast cells as compared to the parental 1,25(OH)2D3. To clarify the mechanism by which nine of these vitamin D3 analogs mediate their remarkably potent biological activities, we have investigated their abilities in PC-3 prostate cancer cells to transactivate a chroramphenicol acetyl transferase (CAT) reporter gene containing a VDRE from the human osteocalcin gene attached to a thymidine kinase minimal promoter. Dose-response studies of Cmpd E showed that in serumless culture conditions, transactivation of the VDRE-CAT was stronger than cmpd J [1,25(OH)2D3]. Then, we investigated the effects of vitamin D3 cmpd J in mice. Our data showed the growth inhibitory action of the vitamin D3 cmpd E in prostate cancer cell line (PC-3) was stastically superior to the non-treatment group in terms of tumor size and tumor weight in mice. In summary, this is the first report of a potent series of 20-cyclopropyl-cholecalciferol vitamin D3 analogs with the ability to inhibit proliferation of LNCaP, PC-3, DU-145, MCF-7 and HL-60 cell lines. These cmpds may mediate their potent anti-proliferative activities through a cell cycle arrest pathway. PMID- 10470103 TI - 1H magnetic resonance spectroscopy evidence that aberrant crypt foci are preneoplastic lesions in the colon. AB - BACKGROUND: Histopathological and genetic studies support the hypothesis that aberrant crypt foci (ACF) represent one of the earliest events in colon carcinogenesis. The purpose of this study is to make use of 1H MRS in conjunction with multivariate methods of analysis to ascertain the validity of the above mentioned hypothesis. MATERIALS AND METHODS: ACF, colonic mucosa and tumor samples taken from thirty-two carcinogen (azoxymethane)-treated Sprague Dawley rats, and of colon mucosa taken from ten healthy animals, were investigated ex vivo by 1H MRS and analyzed using multivariate methods of analysis. RESULTS: The 1H magnetic resonance peak intensities and areas of ACF lie between those from normal and carcinogen- treated mucosa samples and tumors. Multivariate analysis classification of the spectra suggests that the ACF exhibit biochemical characteristics intermediate between the control and AOM-mucosa samples and the tumor groups. CONCLUSION: The use of sophisticated methods of data classification has enabled us to support the hypothesis that ACF represent preneoplastic lesions of the colon. PMID- 10470104 TI - Wortmannin, a phosphoinositide 3-kinase inhibitor, selectively enhances cytotoxicity of receptor-directed-toxin chimeras in vitro and in vivo. AB - BACKGROUND: Generalized resistance of some neoplastic cell lines to treatment with ligand-toxin chimeras has been attributed to an increased rate of lysosomal uptake and degradation following endocytosis of the chimera-receptor complex. Because phosphoinositide 3-kinase (Pl 3-kinase) activity is known to play a role in intracellular trafficking, particularly from endosomes to lysosomes, we hypothesized that co-exposing cells to the Pl 3-kinase inhibitor, wortmannin, might enhance cytotoxicity of ligand-toxin chimeras. METHODS: In vitro, cytotoxicity of five receptor directed-toxin chimeras (bFGF-SAP, bFGF-PE, aFGF PE, HBEGF-SAP, bFGF-gelonin) and an immunotoxin (11A8-SAP) was examined in the presence or absence of this Pl 3-kinase inhibitor against a panel of human neoplastic cell lines: SK-MEL-5 (melanoma), PA-1 (ovarian teratocarcinoma), DU145 (prostatic carcinoma) and MCF-7 (breast carcinoma). In vivo, antitumor activity of a treatment regimen combining wortmannin (1 or 2 mg/kg i.p.) and bFGF-SAP (10 micrograms/kg i.v.) once a week for 4 weeks was evaluated compared to administration of each agent alone in C3H/HeN mice implanted with the FSallC murine fibrosarcoma. RESULTS: At concentrations greater than the reported Ki for Pl 3-kinase inhibition (1-10 microM), wortmannin enhanced cytotoxicity when combined with saporin or gelonin chimeras, but produced subadditive cytotoxicity when combined with Pseudomonas exotoxin chimeras. When low nanomolar concentrations selective for Pl 3-kinase inhibition (5-100 nM) were examined for effects on one receptor directed-toxin chimera, wortmannin dramatically enhanced bFGF-SAP cytotoxicity in three of the four cell lines. A different Pl 3-kinase inhibitor, LY294002 (Ki approximately 1 microM), however, failed to potentiate bFGF-SAP. When administered to mice, wortmannin combined with bFGF-SAP resulted in a significant decrease in tumor volumes compared to vehicle-treated controls that was not observed in mice treated with either agent alone. CONCLUSIONS: Taken together, these results suggest that although wortmannin increases the cytotoxic efficacy of some receptor-directed chimeras, potentiation may occur through an alternative pathway not involving Pl 3-kinase inhibition. PMID- 10470105 TI - Caspase-activation and induction of inducible nitric oxide-synthase during TNF alpha-triggered apoptosis. AB - Activation of the intracellular "death domain" (DD) of the 55kD-TNF alpha receptor by TNF alpha initiates signal and effector cascades with pro- and anti apoptotic function. Co-activation of the adjacent "NO-domain" is followed by induction of inducible nitric oxide-synthase (iNOS) and generation of nitric oxide radicals (NO.). Recently, we have shown NO.-generation to be essential for TNF alpha-induced apoptosis of various tumor cell lines. However, the impact of iNOS activation in relation to other promoters of apoptosis, such as the caspases, is still unclear. Caspase activation, iNOS induction and death rate were therefore investigated in TNF alpha-treated MCF-7 cells. Incubation with TNF alpha (+/- cycloheximide) led to activation of the caspase cascade and was followed by apoptosis. Simultaneously, TNF alpha stimulated induction of iNOS and generation of NO.. Caspase inhibitors DEVD-CHO, YVAD-cmk and YVAD-CHO effectively inhibited caspase activation and prevented apoptosis. Apoptotic cell death was decreased to a similar degree following inhibition of iNOS by L-nitro-arginine methyl-ester (L-NAME). Cell death suppression by caspase inhibition did not result in reduced iNOS activity, as well as L-NAME-dependent prevention of apoptosis was not associated with caspase inactivation. Taken together, TNF alpha induces apoptosis in MCF-7 cells by initiating a two-sided effector pathway including iNOS-induction and activation of caspase 1- and 3-like proteases. Both mechanisms seem to be equally essential for the execution of the death program. The exact nature of their cooperation needs further clarification. PMID- 10470106 TI - Increased neopterin concentrations in patients with cancer: indicator of oxidative stress? AB - In vitro, large amounts of neopterin are produced by human monocytes/macrophages upon stimulation with interferon-gamma. In vivo increased neopterin concentrations in human serum and urine indicate activation of cell-mediated (Th1 type) immune response, e.g., during virus infections, autoimmune diseases, allograft rejection and in certain types of malignancy. In various groups of patients with malignant diseases neopterin concentrations correlate to the stage of disease, and higher neopterin concentrations in serum, urine or ascitic fluid were shown to significantly predict worse prognosis regarding relapse and survival. The amounts of neopterin produced by activated monocytes/macrophages correlate with their capacity to release reactive oxygen species (ROS). With this background, neopterin concentrations in body fluids can be regarded as an indirect estimate of the degree of oxidative stress emerging during cell-mediated immune response. Moreover, recently neopterin was found itself to be capable of enhancing toxic effects induced by ROS. In vitro, neopterin derivatives were able to interfere with intracellular signal transduction pathways involved in, e.g., programmed cell death and the induction of proto-oncogene c-fos or nuclear factor chi B. The data support the view that increased production of ROS--indicated by increased neopterin concentrations--could modulate the development, the proliferation and the survival of malignant cells. PMID- 10470107 TI - Overexpression of cDNA encoding FANCC, SPHAR, MPG, SNM1 or HA 3611 does not render CHO cells more resistant to DNA crosslinking agents. AB - DNA crosslinking agents (DCA) are commonly used cytostatic drugs, whose efficiency in tumor therapy is limited due to the appearance of drug resistant tumor cells. In an effort to modulate the resistance of cells to DCA, we transfected into Chinese hamster cells various cDNAs whose loss of function was previously shown to render cells more sensitive to crosslinking agents. We show that overexpression of FANCC, SPHAR, MPG, SNM1 or HA 3611 (a human homologue of the yeast crosslink DNA repair gene SNM1) does not alter the level of resistance of CHO cells to clinically relevant DCA, such as mafosfamide, melphalan and mitomycin C. Therefore, DCA resistance frequently observed in tumor cells is not likely to be the result of up-regulation of either one of these genes, but a more complex phenomenon. Also, the data suggest that protection of normal cells from toxic side effects of DCA cannot easily be accomplished by transfer of either one of these genes. PMID- 10470108 TI - Metabolite composition in breast tumors examined by proton nuclear magnetic resonance spectroscopy. AB - BACKGROUND: In vivo characterisation of breast tumors using protein (1H) MR spectroscopy relies upon in vitro interpretation of tissue samples. The present study has investigated metabolite composition in extracts from breast tumors and non-involved breast tissue. Multivariate data analysis was used to determinate combinations of metabolites important for differentiation. MATERIALS AND METHODS: Tumor and non-involved breast tissue were obtained from 16 patients undergoing surgical treatment. 1H NMR spectra of perchloric acid tissue extracts were obtained at a BRUKER Avance DRX600 spectrometer. The data was analysed using principal component analysis and probabilistic neural networks. RESULTS: Low levels of glucose and high content of choline compounds were dominant findings in the tumor spectra. Principal component loadings demonstrated this strong association. The spectra were correctly classified using neural network analysis. CONCLUSIONS: Large differences in the metabolite composition of breast tumors and surrounding breast tissues have been documented. PMID- 10470109 TI - Expression of CD 73 (ecto-5'-nucleotidase) in 165 glioblastomas by immunohistochemistry and electronmicroscopic histochemistry. AB - BACKGROUND: CD 73 (5'-nucleotidase) is an ectoenzyme, which is expressed on normal and neoplastic glial plasma membranes. The enzyme binds to intracellular filamentous actin and the extracellular matrix proteins laminin and fibronectin. CD 73 is a signalling pathway metabolite in the immune response of lymphocytes. The ectoenzyme catalyzes the conversion of purine and pyrimidine ribo- and deoxyribo-nucleoside monophosphates (AMP, GMP, IMP) and leads to elevation of the corresponding nucleosides (adenosine) in the extracellular space and might therefore modulate neuronal signalling and vascular perfusion. CD 73 has also been called a cellular motility factor. There is an increasing amount of evidence for the modulatory role of PKC-mediated CD 73 activity in ischemia, regeneration and repair, glioma cell proliferation and a possible invasion promoting feature of the ectoenzyme. The aim of the present study was to investigate the expression patterns of CD 73 together with the labelling of PKC and EGFR. The latter is known as a marker for primary glioblastomas. PATIENTS AND METHODS: We investigated the expression of CD 73 in 165 glioblastoma specimens together with the expression patterns of PKC and EGFR by immunocytochemistry on cryosections with a 4-step grading evaluation by two independent observers. CD 73 was further investigated morphologically by electron-microscopic histochemistry in cell cultures of glioblastoma specimens. RESULTS: With these methods it was possible to demonstrate a dense labelling pattern of glioblastoma specimens with anti-CD 73. 95.7% of the glioblastomas were identified with staining products, 63% with labelling grades 2 and 3. The dense staining of the endoplasmatic reticulum, vesicles, caveolar structures and glial membranes was demonstrated by electron microscopic histochemistry. Some free enzymatic activity was located bound to the ECM components. We observed a significant coexpressions of CD 73 with PKC (p = 0.001) and CD 73 with EGFR (p = 0.022), which is a prospective marker for a high rate of early recurrency. CONCLUSIONS: The CD 73 activity was densely distributed on the membranes of glioblastoma cells in vivo and in cell cultures. The electron microscopic histochemical studies could demonstrate enzymatic activity at the cell membranes and in vesicular structures and caveolae. Free staining deposits located on ECM components may result in a migration- and infiltration-promoting activity. The CD 73 expression could be correlated with the expression grades of PKC and EGFR. The latter has been identified as a prognostic factor which is expressed mainly on primary glioblastomas. PKC is a known tumour metabolite in several proliferation promoting pathways of EGF receptor signalling. PMID- 10470110 TI - Conventional cytogenetics alone is not sufficient for identifying interspecies cell line contamination. AB - Evidence from our laboratory suggests that conventional chromosome banding analysis alone is not sufficient for detecting interspecies contamination of cell lines. To differentiate noncycling interphase cells of murine or other species origin that may be contaminating a human cell line, molecular cytogenetic techniques such as fluorescence in situ hybridization (FISH) with a total human DNA probe must be used. PMID- 10470111 TI - Tumor liberated protein (TLP): its potential for diagnosis and therapy. AB - Our earlier studies led to the purification of a similar antigen from several types of human tumours which had the following properties: It provoked a delayed reaction in certain cancer patients which could be demonstrated both in vivo and in vitro (1). In contrast, healthy individuals did not react to this antigen. Subsequent studies led to a higher degree of purity of tumour antigen which enabled a partial sequence (2). The eight amino acid sequence obtained led to the synthesis of the corresponding peptide. This synthetic peptide was then used to raise specific antibodies in rabbits. The availability of such antibodies enabled to undertake a range of studies. Those investigations revealed that the antibodies were tumour specific because they reacted only with certain types of tumours but not with normal cells nor with other types of tumour tissues. We decided to name this tumour antigen as TLP for tumour liberated protein. PMID- 10470112 TI - Preparation and in vitro evaluation of B-lipiodol as a boron delivery agent for neutron capture therapy of hepatoma. AB - BACKGROUND: We prepared boron containing lipiodol (B-lipiodol), elucidated the retention of B-lipiodol in hepatoma cells and evaluated the in vitro cellular toxicity of B-lipiodol for neutron capture therapy. MATERIALS AND METHODS: Human hepatoma HepG2 cells were used to examine the uptake and retention of B-lipiodol. Light microscopes were used to examine the interaction and retention of B lipiodol globules in individual hepatoma cells. Boron and lipiodol concentrations were determined by inductively coupled plasma-atomic emission spectroscopy and neutron activation analysis, respectively. RESULTS: The boron concentration in B lipiodol drug could reach 2500 ppm. B-lipiodol could be stably retained in serum and culture medium. HepG2 cells appeared proficiently at internalization and persistent retention of B-lipiodol. The boron concentration reached 3.5 micrograms/10(6) cells without approaching saturation at 48 h treatment. CONCLUSION: Hepatoma cells could actively uptake B-lipiodol and a sufficient amount of boron was retained inside the HepG2 cells which could be used for neutron capture therapy. PMID- 10470113 TI - Analysis of microsatellite instability, TGF-beta type II receptor gene mutations and hMSH2 and hMLH1 allele losses in pancreaticobiliary maljunction-associated biliary tract tumors. AB - While pancreaticobiliary maljunctions (PBM) are clearly associated with biliary tract tumor development, little is known about their molecular mechanisms. This study was conducted to assess the contributions of microsatellite instability (MSI), mutations of transforming growth factor type II receptor (TGF-beta RII) and insulin-like growth factor type II receptor (IGF RII) genes and loss of heterozygosity (LOH) of hMSH2 and hMLH1 in 23 biliary tract tumors using PCR methods. MSI was detected by 13 markers in 16/23 samples (69.6%). TGF-beta RII mutations were detected in eight of these (50%), and of the IGF IIR gene in two (12.5%). LOH was detected in 4/16 (25%) at the hMSH2 locus, and 2/16 (12.5%) at the hMLH1 locus. No TGF-beta RII mutations or LOH of hMSH2 and hMLH1 were detected in MSI-negative samples. These findings suggest that MSI plays an important role in carcinogenesis of the biliary tract epithelium with PBM cases. PMID- 10470114 TI - Differential expression of adhesion molecules (CD44, ICAM-1 and LFA-3) in cancer cells grown in monolayer or as multicellular spheroids. AB - Multicellular tumor spheroids have been used to examine numerous aspects of tumor biology since they often recreate the in vivo tumor environment much more closely than other models. Since the three-dimensional organization of cancer cells into spheroids is based upon cell-cell interactions which appear dramatically different in spheroids with respect to monolayer cultures, it can be hypothesized that a modulation in the expression of the molecules which are directly responsible for cell-cell and cell-matrix interactions, particularly the cell adhesion molecules (CAMs), may be involved. In order to test this postulate, the expression of three important CAMs involved in tumor processes (CD44, ICAM-1 and LFA-3) in the human cancer cell lines HT29 (colon adenocarcinoma), A431 (squamous epidermal carcinoma) and A2780 (ovarian carcinoma) grown in monolayer or as multicellular spheroids was compared. The results demonstrate that only two of the lines (HT29 and A431) formed spheroids after six days of gyratory culture while A2780 cells did not form such structures after up to 8 days of culture. In the two cell lines which did form early phase multicellular spheroids, flow cytometric analysis revealed that important differences exist between the same cells grown in monolayer and as spheroids in the quantity of expression of CAMs. PMID- 10470115 TI - Introduction of gadd153 gene into gastric cancer cells can modulate sensitivity to anticancer agents in association with apoptosis. AB - Gadd 153 gene is known as one of the growth arrest and DNA damage inducible genes that may play an important role in signal transduction pathway(s) in response to DNA damage. We have investigated whether the introduction of gadd153 gene into gastric cancer cells could modulate the sensitivity to anticancer agents in association with apoptosis. The transfection of gadd153 gene into MKN45 gastric cancer cells (MKN45gadd153) increased the sensitivity to a variety of anticancer agents, compared to that of neo gene-transfected cells (MKN45neo). The sensitivity to CDDP and VP-16 was increased to a greater extent, whereas the sensitivity to 5-FU and taxotere was increased to a lesser extent. The increase of sensitivity to these drugs was associated with the increase of the formation of internucleosomal DNA ladders in apoptosis. The basal level of gadd153 mRNA was overexpressed in MKN45gadd153 cells, and its induction following the treatment of VP-16 and taxotere was found to a greater extent than that of MKN45neo cells. The analysis of mRNA expression in drug resistance-related genes including mdr1, mrp, topoisomerase II alpha showed that the increase of drug-sensitivity in MKN45gadd153 cells was not due to the changes in expression of drug resistance genes. These results suggest that the introduction of gadd153 gene into gastric cancer cells may modulate the sensitivity to certain anticancer agents by activating AP-1-associated signal transduction pathway(s) leading to apoptosis. PMID- 10470116 TI - Induction of Fas ligand (CD95L) by the toxic mistletoe lectins in human lymphocytes. AB - BACKGROUND: Fas ligand (FasL, CD95L) predominantly expressed on activated cytotoxic T cells and NK cells triggers apoptosis in Fas receptor (Apo-1, CD95) positive target cells. We investigated the expression of FasL, Fas and tumor necrosis factor (TNF) receptor 1 (TNF-R1, CD120a) on cultured human lymphocytes and leukemic T and B cells. MATERIALS AND METHODS: Lymphocytes from six healthy individuals, from four patients with chronic lymphocytic T or B cell leukaemia, and leukemic Molt-4 cells were incubated with the apoptosis- inducing mistletoe lectins (ML I and ML III). RESULTS: Incubation of differentiated lymphocytes with the ML resulted in a significant upregulation of FasL in the surviving CD4+ T helper cells, CD8+ cells and CD19+ B cells. Similarly, the TNF receptor expression increased, while the Fas molecule decreased. In contrast, FasL was not induced in leukemic cells. CONCLUSIONS: Apart from a direct induction of apoptosis in response to an inhibition of protein synthesis by the enzymic ML A chain, ML treatment may indirectly induce apoptosis in Fas+ tumour cells through activated FasL+ lymphocytes. PMID- 10470117 TI - Antitumor efficacy of combination chemotherapy with UFT and cyclophosphamide against human breast cancer xenografts in nude mice. AB - The combination of cyclophosphamide (CPA) and 5-fluorouracil (5-FU) is currently regarded as the most effective therapy for the treatment of patients with advanced and recurrent breast cancer. We evaluated the augmentation of antitumor activity and toxicity by coadministration of CPA and UFT (1M tegafur--4M uracil) instead of intravenous 5-FU on H-31 human breast cancer xenografts in nude mice. The maximum tolerable dose (MTD) of UFT alone (24 mg/kg) and CPA alone (85 mg/kg) had a significant effect on H-31 tumors in mice with 86.6% and 83.0% inhibition rates of tumor growth, respectively, and without loss of body weight, diarrhea or myelosuppression. The combined administration with full and 83.3% MTD of UFT and CPA augmented the antitumor activity compared to that of UFT alone and CPA alone. The relative tumor volume of the UFT plus CPA-treated group to the UFT- and CPA treated groups was 0.28 and 0.36 for the full MTD, and 0.51 and 0.67 for 83.3% MTD, respectively. When CPA was consecutively administered to the tumor-bearing mice for 14 days, there were no decreases in the activities of enzymes related to 5-FU metabolism, but there was an significant increase in the activity of ribonucleotide reductase, suggesting that anabolism of 5-FU derived from tegafur is accelerated to some extent by coadministration of CPA. In conclusion, these results suggest that combination therapy with oral UFT and CPA may be useful for the long-term treatment of cancer patients with advanced and recurrent breast cancers. PMID- 10470118 TI - Evidence that irradiation of far-infrared rays inhibits mammary tumour growth in SHN mice. AB - To evaluate the effect of irradiation of far infrared rays (FIR), the growth of spontaneous mammary tumours of SHN mice was compared among 3 groups: the control was kept until the end of experiment on the normal rack in the absence of FIR and Experimental group I was constantly exposed to FIR. Experimental group Il was raised as the control followed by movement to the FIR rack after mammary tumour appearance. While there was little difference between the control and Experimental group I in mammary tumour growth for 16 days, Experimental group II was significantly lower than the control in this parameter. Furthermore, the percentage of rapidly growing tumours showing greater than 200% of growth rate was apparently lower in Experimental group II. Associated with this, epidermal growth factor receptor expression in mammary tumours, anterior pituitary weight and serum leptin level were significantly decreased in Experimental group II. The findings suggest that whole-body FIR irradiation at ambient temperature could be a possible way of a hyperthermic therapy for tumours. PMID- 10470120 TI - Characterization of dendritic cells generated from peripheral blood of patients with malignant melanoma. AB - Dendritic cells (DCs) are professional antigen presenting cells. Two x 10(7) peripheral blood mononuclear cells (PBMCs) from each of 6 melanoma patients and 4 normal donors were incubated in 6-well plates. The adherent cells were cultured in GM-CSF (800 U/ml) and IL-4 (1000 U/ml) for 10 days. Anti-CD80, anti-CD86, anti HLA-DR, anti-HLA-class I monoclonal antibodies and supernatant from two melanoma cell lines were incubated with DCs before responder T cells were added in mixed lymphocyte reaction (MLR). The mean yields of DCs were 3.6 x 10(5) and 9.3 x 10(5) for donor and the melanoma patients respectively. MLR was inhibited by anti CD80, anti-CD86, anti-HLA-DR, anti-HLA-Class I and supernatant of melanoma cultures. The characteristics of DCs generated from melanoma patients were comparable to those from donors. DC function was inhibited by soluble factors in melanoma cultures. Further studies are warranted to characterize these inhibitory factors. PMID- 10470119 TI - New genes potentially involved in breast cancer metastasis. AB - Identification of new genes involved in the pathogenesis of breast cancer opens new avenues for improved diagnostic markers and new molecular targets for improved treatment of this malignancy. In the following we review genes with proved involvement in invasion and metastasis of breast cancer as well as genes which exhibit an expression pattern that correlates with invasion and metastasis. PMID- 10470122 TI - G2/M checkpoint is p53-dependent and independent after irradiation in five human sarcoma cell lines. AB - To determine the role of p53 in G2/M arrest, G2/M transition and apoptosis, we investigated five human sarcoma cell lines with different p53 gene status in their response to X-rays. The p53 status of the cell lines was mutant (US 8-93, LMS 6-93 and RD), null (SAOS-2) and wildtype (A-204). Clonogenic survival of the cell lines varied as the survival fraction at 2 Gy (SF2) ranged from 0.28 to 0.79. Compared with the mutated p53 cell lines (SF2 with a range from 0.46 to 0.79) the clonogenic survival of the wildtype p53 (wt-p53) cell line A-204 (SF2 = 0.34) was lower. The p53 null cell line (SAOS-2) was also sensitive to X-rays (SF2 = 0.28). We detected, in all cell lines a similar behavior in their response to irradiation with G2/M arrest and apoptosis. However, the maximal rate of apoptosis with a range from 7.0 to 18.0% was rather small. The decrease of G2/M cells was coupled with an increased percentage of apoptotic cells. However, a different delay in G2/M did not result in a change of radiation sensitivity. Western analyses showed an increased P53 level only for the cell line A-204 (wt p53) after irradiation. Our results point out that there is not always a simple relationship between p53 gene status and radiation sensitivity. We suggest, that wt-p53 plays an active role in G2/M arrest and in decreasing the number of G2/M cells as a response to apoptosis. Therefore, p53-dependent regulation in G2/M may be as important as p53-independent mechanisms. PMID- 10470121 TI - Replication error in colorectal carcinoma: association with loss of heterozygosity at mismatch repair loci and clinicopathological variables. AB - Instability of microsatellite DNA or replication error (RER) is characteristic of tumours caused by mismatch repair (MMR) deficiency. Germline mutations in MMR genes are associated with Hereditary non-polyposis colorectal carcinoma (HNPCC) and somatic mutations in these genes are also found in a substantial fraction of colorectal cancers (CRC). In this study we concurrently screened colorectal tumours for the RER phenotype and loss of heterozygosity (LOH) at MMR gene loci. The RER phenotype was evident in 47/197 (24%) tumours. RER was more commonly detected in young patients (< 50 years) and in tumours located in the proximal colon. RER was positively associated with LOH at the hMSH2/hMSH6 loci on chromosome 2p, where LOH was observed in 46% of the RER+ tumours. LOH at hMLH1 and hPMS1 loci was more frequent in the younger patients (< 50 years). RER was not associated with clinicopathological parameters, such as Duke's stage and tumour differentiation (grade). The RER phenotype was associated with better overall survival, but there was a trend towards significance when multivariate analysis was used. This indicates that loss of MMR genes generate a less aggressive phenotype, and raises the question about RER being a useful indicator of prognosis for CRC patients. PMID- 10470123 TI - Cytokeratin 8 and 18 fragments measured in serum and their relation to survival in patients with non-small cell lung cancer. AB - BACKGROUND: In this study we investigated if the newly developed monoclonal antibodies against Cytokeratin 8 and 18 fragments (Cyk 8/18) have prognostic information in patients with non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Serum from 69 patients with NSCLC was investigated using a sandwich ELISA, Cyk 8/18, provided by IDL Biotech, Sollentuna Sweden. RESULTS: Cyk 8/18 levels varied between 0.34-14.2 ng/mL, compared with a cut-off value of 1.0 ng/mL for healthy individuals (95% specificity). Using that cut-off value, 80% of NSCLC patients had elevated levels. A statistically significant diminished survival was found for Cyk 8/18 values of 8.0 ng/mL or higher (p = 0.0001). When survival data and Cyk 8/18 levels were analysed according to continuous Cox regression analysis, increased levels of Cyk 8/18 were significantly related to decreased survival (p = 0.016). CONCLUSIONS: The Cyk 8/18 monoclonal antibody had in this study prognostic information regarding survival in patients with NSCLC. PMID- 10470124 TI - Antitumor activity of benzo[a]phenothiazines. AB - We have previously reported on the diverse biological activities of benzo[a]phenothiazines, such as the induction of antitumor and antimutagenic activity in vivo, and differentiation and apoptosis in vitro. The relationship of radical generation and pi-spin density or dipole moment was investigated, using quantum-chemical calculation with UHF/PM3. These data suggest that the origin of radical generation by active benzo[a]phenothiazines, which affect such biological activities might be N-atom at position 12. PMID- 10470125 TI - Cell cycle inhibitory protein p27 in esophageal squamous cell carcinoma. AB - BACKGROUND: p27 protein is one of the cdk inhibitors which regulates the progression from G1 to S phase of the cell cycle. Reduced expression of p27 protein has been reported to be correlated with poor clinical outcome in patients with various cancers. MATERIALS AND METHODS: We performed immunohistochemistry of both p27 and Ki67 in 136 cases of resected human esophageal squamous cell carcinoma, and evaluated the association between p27 immunoreactivity and clinicopathological features including clinical outcome in these patients. We also examined the correlation between labeling index or the percentage of positive tumor cells for p27 and Ki67 in serial tissue sections in these cases. RESULTS: Cases with invasion of the muscularis propria or adventitia had significantly (p < 0.05) higher p27 LI (65.0 +/- 23.7) than those with invasion limited to mucosa or submucosa and those with carcinoma in situ (58.9 +/- 18.3). There were no significant correlations between p27 and other clinicopathological factors such as sex, age, tumor size, differentiation type, nodal status and histological stage. The cases with p27 LI below 40% tended to have a worse prognosis than those with p27 LI above 40%. There was no significant correlation between Ki67 and p27 LIs. CONCLUSIONS: Reduced expression of p27 may be correlated with the biological behavior of esophageal squamous cell carcinoma and may play an important role in the early stages of cancer. PMID- 10470126 TI - A comparison of the anticancer activities of dietary beta-carotene, canthaxanthin and astaxanthin in mice in vivo. AB - The anticancer activities of beta-carotene, astaxanthin and canthaxanthin against the growth of mammary tumors were studied in female eight-wk-old BALB/c mice. The mice were fed a synthetic diet containing 0, 0.1 or 0.4% beta-carotene, astaxanthin or canthaxanthin. After 3 weeks, all mice were inoculated with 1 x 10(6) WAZ-2T tumor cells into the mammary fat pad. All animals were killed on 45 d after inoculation with the tumor cells. No carotenoids were detectable in the plasma or tumor tissues of unsupplemented mice. Concentrations of plasma astaxanthin (20 to 28 mumol/L) were greater (P < 0.05) than that of beta-carotene (0.1 to 0.2 mumol/L) and canthaxanthin (3 to 6 mmol/L). However, in tumor tissues, the concentration of canthaxanthin (4.9 to 6.0 nmol/g) was higher than that of beta-carotene (0.2 to 0.5 nmol/g) and astaxanthin (1.2 to 2.7 nmol/g). In general, all three carotenoids decreased mammary tumor volume. Mammary tumor growth inhibition by astaxanthin was dose-dependent and was higher than that of canthaxanthin and beta-carotene. Mice fed 0.4% beta-carotene or canthaxanthin did not show further increases in tumor growth inhibition compared to those fed 0.1% of each carotenoid. Lipid peroxidation activity in tumors was lower (P < 0.05) in mice fed 0.4% astaxanthin, but not in those fed beta-carotene and canthaxanthin. Therefore, beta-carotene, canthaxanthin and especially astaxanthin inhibit the growth of mammary tumors in mice; their anti-tumor activity is also influenced by the supplemental dose. PMID- 10470127 TI - Effects of docetaxel on the in vitro growth of human myeloid progenitors. AB - Haemotoxicity is usually the primary and dose-limiting side-effect of docetaxel (TXT) a semysyntetic analogue of paclitaxel which has acquired an important role in anticancer treatment. This research presents the results of an in vitro toxicity study of TXT on myeloid progenitors obtained from healthy volunteers and assayed as CFU-GM. Peripheral blood mononucleated non-adherent cells (MNAC) were incubated for 24 h at standard conditions with increasing concentrations of TXT and then cultured for CFU-GM assay. At every concentration severe CFU-GM growth inhibition was observed. In a second set of experiments MNAC were sequentially exposed to TXT and then to doxorubicin or cisplatin or vinorelbine or etoposide at appropriate concentrations. In a third set the sequence of exposure was reversed. No difference of CFU-GM growth inhibition was observed between the alternate sequences. These findings suggest that the toxicity on CFU-GM in vitro growth of TXT combinations with other anticancer drugs is sequence-independent. PMID- 10470128 TI - Chemical structure and tumor type specificity of "half-mustard type" phenothiazines. AB - The antiproliferative activity of six "half-mustard type" phenothiazines against a total of 54 tumor cell lines: 4 leukemia, 9 non-small-cell lung, 7 colon-, 5 CNS-, 8 melanoma, 6 ovarian-, 8 renal-, 1 prostate and 6 breast cancer was determined by NCI-Information Intensive-Approach. The C-2 position of phenothiazines were substituted with H, Cl and CF3 groups. The half-mustard and ring system was linked either by a propylene or a butylene bridge. Colon-cancer cell showed the highest sensitivity against "half-mustard type" phenothiazines, followed by leukemia, melanoma, prostate-, CNS-, breast-, lung-, renal and ovarian cancer cells. These data suggest the "cancer-type-specific" antitumor action of "half-mustard type" phenothiazines. PMID- 10470129 TI - Antiproliferative and apoptosis-inducing effects of an antitumor glycoprotein from Streptococcus pyogenes. AB - An acidic glycoprotein (SAGP) purified from an extract of Streptococcus pyogenes has been shown to inhibit the growth of methylcholanthrene-induced fibrosarcoma A (Meth A) cells via pertussis toxin-sensitive GTP-binding protein. The present study revealed that SAGP has activity to induce apoptosis in Meth A cells as assessed by DNA fragmentation and cell morphology with chromatin staining. The SAGP-induced DNA fragmentation in Meth A cells was augmented by herbimycin A, an inhibitor of protein tyrosine kinase, and prevented by orthovanadate, an inhibitor of protein tyrosine phosphatase. The growth inhibitory effect of SAGP on Meth A cells was reduced by orthovanadate, whereas the effect tended to be increased by herbimycin A. Western blotting analysis using antiphosphotyrosine antibody demonstrated that tyrosine phosphorylation of 170 kDa cellular protein was diminished in the cells incubated with SAGP. The inhibition of protein tyrosine phosphorylation was neither observed in the cells incubated with SAGP and orthovanadate nor in the cells incubated with heat-inactivated SAGP. These findings indicate that inhibition of tyrosine phosphorylation by protein tyrosine phosphatase(s) may be responsible for the SAGP-induced apoptosis and inhibition of cell growth. PMID- 10470130 TI - Topoisomerase II alpha expression in esophageal squamous cell carcinoma. AB - BACKGROUND: DNA topoisomerase II alpha (topo II alpha) is associated with active cell proliferation of mammalian cells. Topo II alpha overexpression has been reported in a number of human malignancies and is considered to be related to their biological behaviors. MATERIALS AND METHODS: We examined the expression of topo II alpha immunohistochemically in 136 cases of human esophageal squamous cell carcinoma, 10 foci of squamous dysplasia and 10 non-pathologic squamous epithelium. We calculated the labeling index (LI) or the percentage of immunopositive cells for Topo II alpha and Ki67, and Topo II alpha LI/Ki67 LI (T/K ratio). These findings were then correlated with clinicopathological features of the patients including their clinical outcome. RESULTS: Both topo II alpha and Ki67 immunoreactivity were detected in the nuclei. A significant positive correlation was obtained between Topo II alpha and Ki67 LIs in all the specimens examined. Topo II alpha LI and T/K ratio were 24.5 +/- 8.0% and 1.04 +/ 0.64 for carcinoma, 19.1 +/- 15.2% and 0.68 +/- 0.29 for dysplasia and 14.0 +/- 14.1% and 0.55 +/- 0.17 for non-pathologic epithelium, respectively. Topo II alpha LI and T/K ratio in carcinoma cases were significantly higher than those of normal epithelium. Topo II alpha LI alone did not correlate with any of clinicopathological parameters examined but among carcinoma cases, cases with lymph nodes metastasis or higher histological stages had significantly higher T/K ratio than those without lymph node metastasis or lower histological stages. In addition, carcinoma cases with T/K ratio of greater than 0.8 demonstrated significantly worse prognosis than those with T/K ratio of smaller than 0.8. CONCLUSIONS: The relative overexpression of topo II alpha as compared with Ki67, i.e., increased T/K ratio was detected in esophageal squamous cell carcinoma and is considered to represent a dysregulation or qualitative alteration in topo II alpha, possibly associated with malignances, as reported in other human cancers. In addition, topo II alpha overexpression may also be correlated with the aggressive biological behavior of the patients with esophageal squamous cell carcinoma. PMID- 10470131 TI - Antiproliferative effects of a product isolated from the gorgonian Rumphella aggregata. AB - A fraction isolated from the gorgonian Rumphella aggregata (Plexauridae) was studied vitro on asynchronous cells of a human non-small-cell-bronchopulmonary carcinoma line (NSCLC-N6). Cell growth appeared to be inhibited in the Gl phase of the cell cycle, and kinetic studies in pretreated cells showed that this growth arrest was irreversible. These events seem to show a terminal maturation induced by this new product. PMID- 10470132 TI - Inhibitory potential of Chlorella vulgaris (E-25) on mouse skin papillomagenesis and xenobiotic detoxication system. AB - The present study assesses the modulatory potential of Chlorella vulgaris (E-25) on murine skin papillomagenesis, and the role of xenobiotic detoxication system in modulating the papillomagenesis pattern. Topical application of E-25 (500 mg/kg b.w./day) during peri-, post- or peri- and post-initiational stages of 7,12 dimethylbenz [a] anthracene (DMBA)-induced papillomagenesis, significantly modulated the a) tumor burden to 5.00, 4.33 and 3.94 (positive control value: 5.88 b) cumulative number of papillomas to 90, 78 and 67 (positive control value: 106); and c) percent incidence of mice bearing papillomas to 94, 90 and 89 respectively (positive control value: 100). E-25 treatment alone or during peri-, post- or peri- and post-initiational stages significantly elevated the sulfhydryl (-SH) and glutathlone S-transferase (GST) levels in the liver and skin tissues. However, the levels of microsomal cytochrome b5 (Cyt. b5) and cytochrome P-450 (Cyt. P-450) were not appreciably modulated by the topical treatment of E-25. The results suggest the chemopreventive potential of E-25 during peri-, post- or peri and post-initiational stages of murine skin papillomagenesis. The possible significance of xenobiotic detoxication system in modulating the papillomagenesis pattern is discussed. PMID- 10470133 TI - Aberrant p16 expression is correlated with hemizygous deletions at the 9p21-22 chromosome region in non-small cell lung carcinomas. AB - The p16 protein is encoded by the CDKN2 gene, and functions as an inhibitor of cyclin-dependent kinase 4 and 6 (CDK4/6). Phosphorylation of the retinoblastoma protein (pRb) by CDK4/6 represents a vital step in cell cycle progression. Alterations of p16INK4A are frequent events in human malignancies. In non-small cell lung carcinoma (NSCLC) the data concerning the mechanisms of p16INK4A inactivation suggest that point mutations and aberrant methylation of its promoter can only account for a proportion of the cases with abnormal p16 immunoexpression. The role of deletions in this procedure is not yet clarified. In order to gain more insight into the role of deletions in p16INK4A deregulated expression, we investigated the state of the chromosomal region 9p21-22 in a series of 57 NSCLCs, by performing a detailed mapping analysis, using a tight cluster of highly polymorphic microsatellite markers, and correlating the findings with p16 immunostaining. Abnormal p16 expression was observed in 46% of the NSCLCs examined. No relationship was observed between p16 abnormal staining and various clinicopathological parameters. Abnormal p16 protein staining was strongly associated with hemizygous deletions at the IFNA and D9S171 microsatellite loci, which demarcate the region encoding the p16INK4A gene (P = 0.002). These findings suggest that deregulated expression of p16 is involved in the multistage process of NSCL carcinogenesis and that deletions may represent a predominant mechanism of p16INK4A inactivation. A significant percentage also of LOH was noticed at the D9S162 (35%) and D9S126 (38%) loci which lie 6cM and 4cM, respectively, far from the area which encodes p16INK4A, implying that other tumor suppressor genes (TSGs) may reside in this region. Although the overall incidence of LOH at the examined region was high (58%), we did not observe any correlation with smoking habits, histology and lymph node status. Another noteworthy finding was the existence of microsatellite instability (MI) in 11% of the patients. MI provides a marker for replication error phenotype (RER+), a recently defined manifestation of genetic instability observed in a wide range of tumors. In conclusion, alterations (LOH + MI) at the 9p21-22 chromosome region are frequent events in NSCLCs and may affect directly or indirectly the expression of p16. PMID- 10470134 TI - A novel human malignant fibrous histiocytoma cell line of the heart (MFH-H) with secretion of hematopoietic growth factors. AB - Primary malignant fibrous histiocytoma (MFH) exhibits an extremely adverse prognosis. Investigations into the principles determining the biological aggressiveness of this cardiac tumor would be facilitated by an appropriate in vitro model. Therefore, we report on the first permanent cell line (MFH-H), derived from a human cardiac MFH. The original tumor had shown coexpression of cytoskeletal filaments typical of mesenchymal (vimentin), epithelial (cytokeratins) and neurogenic (neurofilaments) differentiation. This potential for multidirectional differentiation was observed in the MFH-H cell line as well and indicated marked plasticity of gene activation acquired during the process of neoplastic transformation. Pronounced genetic alterations also became evident from cytogenetic analysis, which revealed a highly variant karyotype with multiple numeric and structural chromosomal aberrations. Secretion of G-CSF, GM CSF and M-CSF was shown to be another feature of deregulated gene expression in MFH-H cells. Direct autocrine effects of their hematopoietic growth factors, however, were precluded by the lack of the corresponding receptors. In conclusion, the cell line MFH-H will provide an appropriate in vitro model to analyze the biological properties of this cardiac malignancy in more detail, especially with regard to a possible immunomodulating capacity of MFH-derived hematopoietic growth factors. PMID- 10470135 TI - Induction of apoptosis in neoplastic cells by anthracycline antitumor drugs: nuclear and cytoplasmic triggering? AB - In spite of extensive investigation, the mechanism for cell cytotoxicity of the anthracycline antitumor drug adriamycin (ADR) has not yet been completely understood but the nature of the cytotoxic effects of this drug is generally related to its interaction with nuclear components, such as DNA and topoisomerase II. In a previous paper, we studied, using Confocal Laser Scanning Microscopy (CLSM), the localization of ADR and its glutathione (GSH)-conjugate (ADRIGLU), obtained by the anaerobic reaction of the parent anthracycline with reduced GSH, in drug-sensitive and in multidrug resistant (MDR) cells. In all drug-sensitive lines used, ADR was mostly located in the nuclei, while its GSH-conjugate was found only in the cytoplasm, predominantly in the Golgi region. In this study we examined the morphological changes induced by ADR or its GSH-conjugated adduct (ADRIGLU) treatments in TVM-A12 (clone 2) melanoma and K562 erythroleukemia human cell lines, correlated to programmed cell death (apoptosis). We observed that ADR induced apoptosis in both cell lines tested after 5 h treatment: CLSM and Scanning Electron Microscopy (SEM) showed cell shrinkage, fragmentation and condensation of nuclear chromatin, cell surface blebbing and cytoplasmic vacuolization. On the contrary, ADRIGLU-induced fragmentation and condensation of nuclear chromatin, typical of apoptosis, only after 48-72 h treatment. Cytoflourimetric assay by propidium iodide staining confirmed the data obtained by CLSM and SEM. Our data suggest that apoptosis activation by anthracycline antitumor drugs is induced not only by direct interaction with nuclear components but also with cytoplasmic compartments. PMID- 10470136 TI - Up-regulation of growth hormone receptor immunoreactivity in human melanoma. AB - Growth hormone (GH) exerts its regulatory functions in controlling metabolism, balanced growth and differentiated cell expression by acting on specific receptors, which trigger a phosphorylation cascade resulting in the modulation of numerous signalling pathways, and dictate gene expression. Immunohistochemical techniques were used to demonstrate the presence of growth hormone receptors in 126 formalin-fixed, paraffin-embedded melanocytic tumours comprising melanocytic naevi, superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma and metastatic melanomas. The relative proportion of positive cells and intensity of staining was higher in neoplastic cells, compared to normal cutaneous cells. Of the 76 cases of common melanocytic naevi (CMN) studies, 46 were weakly reactive with MAb 263. Heterogeneity of immunoreactivity was found in primary melanoma lesions with a variable range of positive cells. Of 37 cases studied, 34 were moderately to strongly positive. Immunoreactivity showed subcellular localization of the GH-receptor in cell membranes, was predominantly cytoplasmic, but strong nuclear immunoreaction was also apparent in many instances. The nuclear localization of immunoreactivity is the result of nuclear GH receptor/binding protein, identically to the cytosolic and plasma growth hormone binding protein. Intense immuno-reactivity was also observed in the cellular Golgi area of established cell lines and cultured tissue-derived cells in exponential growth phase, indicating cells are capable of GH-receptor synthesis. In the primary lesions, dermal tumour cells tended to be more immunoreactive relative to those seen in the dermal region. Metastatic lesions in various organs also expressed growth hormone receptors in secondary tumour cells and all of the metastatic cases were positive. The expression of GH-receptors in human melanoma cells means that these cells are directly responsive to GH action and that GH may stimulate local production of IGF-I, which then acts in an autocrine mechanism. PMID- 10470137 TI - Establishment and characterization of human immature teratoma cell line (TES-1). AB - A new human immature teratoma cell line, TES-1, was established from a surgical specimen from a 12-year old male with third ventricular immature teratoma. TES-1 shows polygonal morphology rich in neurites, and proliferated as adherent monolayer, with an approximate population doubling time of 48 hours. Electron microscopic analysis revealed the presence of swollen rough endoplasmic reticulum, and prominent lipid droplets, lysosomes and microfilaments. The chromosome numbers were between 41 and 160 (mode 78), including abnormal karyotypes 1p-, 5q-, 12p+ and 17p+ (G-band analysis). Hetero-transplantation of TES-1 into BALB/c nude mice produced no visible tumors. Multipotential differentiation was not induced in TES-1 monolayer culture, but significant neuron specific enolase activity was expressed in both extracellular (by RIA method) and intracellular fractions (by immunohistochemical method), suggesting the differentiation toward neurocytes. This cell line provides a useful in vitro model for the pathophysiological analysis of immature teratoma. PMID- 10470138 TI - Xenotransplantability of human squamous cell lung cancer in nude mice is not affected by angiogenic factors. AB - Sixty-two human squamous cell lung carcinomas were analyzed for expression of various angiogenic growth factors and their receptors using immunohistochemistry. The data were correlated with xenotransplantability of these tumors in nude mice. None of the factors investigated did show an association with xenotransplantability. However, there was a trend that specimens lacking VEGF165 were established as xenografts at a higher incidence (52%) than those expressing VEGF165 (39%). PMID- 10470139 TI - Spectrum of extracellular matrix degrading enzymes in normal and malignant T lymphocytes. AB - Human T cells produce and release fibronectin degrading neutral serine proteases with a molecular weight of 50 kD, 70-80 kD (doublet) and 95 kD and have a cell associated 400 kD fibronectin degrading enzyme. In addition, human T cells produce proteases with m.w. 50, 70-80 kD and 400 kD which degrade laminin. CD 4+ T lymphocytes from a non-malignant cloned human T cell line produce a 92 kD gelatinase (MMP 9) and malignant T cell lines release, in addition to the 92 kD gelatinase, low amounts of a 72 kD gelatinase (MMP 2). Purification of the enzymatic activities using benzamidine sepharose yields a 50 kD and a 70 kD band of which the 50 kD band has fibronectin degrading capacity. The purified enzymes do not react with monoclonal antibodies to various previously characterized proteolytic enzymes present in T cells. T lymphocytes from a non-malignant cloned human T cell line produce high amounts of the 50 and 70-80 kD proteases directly after stimulation with anti-CD 3 monoclonal antibodies whereafter the production of these enzymes declines with time. The expression of the 400 kD fibronectin degrading protease is downregulated by crosslinking of alpha 4 beta 1-integrin receptors on T cells using monoclonal antibodies. Thus, T lymphocytes produce several matrix degrading enzymes with multiple substrate specificities. The expression of these enzymes is controlled partly by lymphocyte activation signals or by direct signalling via beta 1-integrins. PMID- 10470140 TI - Antioxidant enzyme defence responses of human MCF-7 and SW480 cancer cells to conjugated linoleic acid. AB - The relationship between antioxidant-enzyme defence responses and cellular growth suppression in human MCF-7 (breast) and SW480 (colon) cancer cells, exposed to CLA in culture was studied. MCF-7 and SW480 cells (1 x 10(6)/flask) were cultured in appropriate medium for 4, 8 and 12 days with varying levels of CLA (0-30 ppm). A dose-dependent decrease in cell numbers and increase in lipid peroxidation, as determined by thiobarbituric acid reactive substances (TBARS) was observed in both cell lines following incubation with CLA. Exposure of both cell lines to 20 ppm CLA for 2-6 days produced a reduction (83-91%) in 3H-leucine incorporation into protein while 3H-uridine and 3H-thymidine incorporation into RNA and DNA were reduced by 49-91% and 86-98%, respectively, compared with untreated control cells. The activities of superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) were induced in both cell lines exposed to CLA (20 ppm) over a period of 12 days, although to a greater extent in MCF-7 cells than in SW480 cells. The data indicate that CLA-induced cytotoxicity against MCF-7 and SW480 cancer cell lines is related to the extent of lipid peroxidation of CLA treated cells and affirm that the CLA- induced antioxidant enzymes failed to protect these cells from cytotoxic lipid peroxidation products. PMID- 10470141 TI - Alterations of tumour cells, stroma and apoptosis in rat prostatic adenocarcinoma following treatment with histamine, interleukin-2 and irradiation. AB - The present study was designed to determine whether IL-2 and histamine alone, or in combination could modulate the effects of irradiation on Dunning (R3327) rat adenocarcinomas at the cellular level. Copenhagen x Fisher rats carrying bilateral tumours in the flank were used. When the tumours had a median volume of 150 mm3, one group of rats was treated with histamine alone (4 mgkg-1 subcutaneously on week days), another group with interleukin-2 (IL-2) alone (425 IU kg-1 continuous infusion) and a third group with both histamine and IL-2 during 6 weeks. Irradiation was given one week after the onset of treatment with histamine and/or IL-2, with a linear accelerator 6 MV, in a dose of 6 Gy/day for 3 days to the tumour of one side, while the other side served as control. Morphometric analyses of the amount of cystic structures, volume density for tumour epithelium, stroma and acinar lumina, the number of activated macrophages and natural killer cells (NK-cells) and in situ detection of apoptotic cells was carried out 5 weeks after the irradiation, when the experiment was ceased. The combination of IL-2 with histamine and irradiation significantly augmented the reduction of tumour cells (p < 0.002) and increased the number of apoptotic cells (p < 0.007) compared to irradiation alone. The number of cystic structures within tumour tissue increased in all tumours that received histamine, but was most pronounced in the three combination group. A strong negative correlation between the epithelial cells and the apoptotic index (rs = -0.61, p < 0.0001) and a strong positive correlation between the stroma and the apoptotic index (rs = 0.59, p < 0.001) was found. A prominent infiltration of activated macrophages was observed in the irradiated group. This infiltration was impaired by the drugs. The results suggested that the used three modality treatment could be of value in increasing the efficacy of local radiotherapy concomitantly with a most plausible effect on micrometastatic spread. The results also propose that volume measurements alone are not an optimal parameter when evaluating effects of new treatment modalities. PMID- 10470142 TI - Induction of histone acetylation and growth regulation in eryrthroleukemia cells by 4-phenylbutyrate and structural analogs. AB - The objective of this investigation was to study the relationship between histone acetylation and growth inhibition by 4-phenylbutyrate and structural analogs. Inhibition of growth of DS19 mouse erythroleukemia cells and K562 human leukemic cells by 4-phenylbutyrate did not appear to be mediated by glutamine depletion. Vanadate blocked differentiation of DS19 cells but did not affect the hyperacetylation of histones. 2-phenylbutyrate was a more effective inhibitor of cell proliferation than 3-phenylbutyrate but was less effective as an inducer of histone acetylation. 4-Phenylbutyrate was a more effective inhibitor of histone deacetylase and inducer of histone acetylation than the structural analogs examined including 2- and 3-phenylbutyrate, cinnamate, methoxycinnamate, 2 phenoxybutyrate and phenoxyacetate. PMID- 10470143 TI - Expression of CD44 isoforms in human breast carcinoma xenografts is not influenced by the treatment of mice with cytostatics or (anti-)hormones. AB - CD44 standard (s) and variant (v) isoforms have been discussed to be implicated in progression and metastasis of different malignomas. For breast carcinomas, the results of different studies are contradictory. These apparent discrepancies suggest that CD44 isoforms are not available on the tumour cell surface, but could be regulated by different endogenous and exogenous factors. Here we report the regulation of CD44 isoforms in xenografted breast cancer cell lines by cytostatics, hormones and antihormones. The human breast cancer models MDA-MB 435, MCF-7, NCI/ADR, 4296, 4151 and 4134 were transplanted into the mammary fat pad of nude mice. When tumours reached a palpable size, animals were treated with farmorubicine, cyclophosphamide, estradiol, tamoxifen or progesterone, respectively. At different times after treatment, serum and tumours were taken. The expression of CD44 and its isoforms was determined by immunohistochemistry and RT-PCR, serum levels were measured by human specific ELISA kits. Serum levels of CD44s and v6 varied among the tumours. For 3/6 tumours we found differences between control groups and treated animals. Immunohistochemical results remained unchanged: each tumour showed a specific pattern of CD44 expression, but this pattern did not change when the animals received cytostatics, hormones or antihormones. The same held true for RT-PCR-results. Also, the time of tumour collection had no influence on CD44 expression. Therefore, it can be concluded, that in the xenografted breast cancer cell lines a regulation of CD44 isoforms by farmorubicine, cyclophosphamide, estradiol, progesterone or tamoxifen could not be found, while serum levels were influenced in some cases probably due to tumour cell kill and shedding of surface proteins into blood stream. PMID- 10470144 TI - Application of high energy shock waves to cancer treatment in combination with cisplatin and ATX-70. AB - BACKGROUND: The anti-tumor effects of high-energy shock waves (HESW) in combination with cisplatin (CDDP) and ATX-70, as a photosensitizer, were investigated in vitro and in vivo. METHODS: Active oxygen generation by HESW was studied by means of electron spin resonance (ESR) system. The anti-tumor effects of HESW in combination with cisplatin (CDDP) and ATX-70 was investigated in vitro and in vivo. RESULTS: HESW exposure induced singlet oxygen or hydroxyl radical. Adding ATX-70 enhanced the cell damage induced by HESW dose-dependently. Ascorbic acid inhibited the combined effect of HESW and ATX-70. Adding CDDP did not enhanced cell damage by HESW. The combined therapy of HESW, CDDP and ATX-70 showed further inhibition of tumor growth in vivo. CONCLUSIONS: These results indicate that HESW activated ATX-70, and that the anti-tumor effect of HESW and ATX-70 was caused by active oxygen generation. HESW is able to be utilized with any other cancer treatment. PMID- 10470145 TI - P-glycoprotein (Pgp) does not affect the cytotoxicity of flavonoids from Sophora flavescens, which also have no effects on Pgp action. AB - Sophoraflavanone, kurarinone (GS08), norkurarinol (GS11), kurarinol (GS12) and kushenol K are cytotoxic flavonoids isolated from Sophora flavescens. In this study, we tested the cytotoxicity of those flavonoids to human cancer cells including P-glycoprotein (Pgp)-expressing HCT15 cells and its multidrug resistant subline, HCT15/CL02 cells. HCT15/CL02 cells revealed resistance to GS08, GS11 and GS12 about 2 fold in comparison with HCT15 cells. Nonetheless, verapamil, a Pgp inhibitor, could not increase the cytotoxicity of all the flavonoids tested. We also investigated that the flavonoids could modulate the Pgp action. At nontoxic concentrations, the flavonoids could not effect on the cytotoxicity of paclitaxel, a well-known Pgp-substrate. The flavonoids also had no effects on the accumulation of rhodamine 123 in all the cells tested at 10 microM. From the results, we concluded that Pgp had no effect on the cytotoxicity of the flavonoids, and the flavonoids also had no effect on the action of Pgp. Our results also suggested that HCT15/CL02 cells had additional mechanisms for drug resistance distinct from Pgp overexpression. PMID- 10470146 TI - Tissue distribution and metabolism of gamma-linolenoyl-3-eicosapentaenoyl propane diol enterally or intravenously administered to mice bearing human pancreatic carcinomas. AB - Synthetic propane diol lipids have been proposed as novel compounds to deliver cytocidal polyunsaturated fatty acids (PUFA) such as gamma-linolenic (GLA) and eicosapentaenoic (EPA) acids. To assess the biodistribution and metabolism of these PUFA in immunodeficient mice bearing human pancreatic carcinomas (AsPC-1), gamma-linolenoyl-3-eicosapentaenoyl propane diol (GE diol) was provided in a fat free diet (5% w:w) for 6 weeks or parentally administered as 14C-GE diol (1 or 3 consecutive doses of 1.66 g/kg/day) in an innovative non-ionic digalactosyldiacylglycerol emulsion. In tumor, liver, brain, kidney, plasma and fat tissue of mice fed GE diol, PUFA were increased over 25-fold, except for arachidonic acid (AA) levels, which were reduced or remained constant when compared to mice fed control corn oil diet. GLA and EPA were mainly stored in fat tissue. The recovery of radioactivity from the i.v. infected 14C-GE diol was dose and time dependent. Ten days after the i.v. infusion, GLA was only detected in substantial concentrations in tumor and in fat tissue (21 and 202 micrograms/g, respectively). Overall, these studies showed that: GE diol emulsions provide 640 fold higher doses of both GLA and EPA without causing hemolysis or adverse effects in the host mouse when compared to free PUFA infusions; GE diol is metabolized after oral or i.v. administration; tumor concentrations of GLA and EPA from the enterally administered diol were 4 to 13-fold higher than the in vitro cytotoxic levels; EPA, competes with AA and probably inhibits the activity of delta 5 desaturase without affecting the elongation of GLA in the host and tumor tissue; the change in PUFA profile modifies the substrates for eicosanoid synthesis. In short, a potentially desirable cytotoxic PUFA pattern can be achieved in host tissues and, in particular, in a human pancreatic tumor by providing GLA and EPA in the form GE-diol. These findings guarantee further investigations in oncology with this neutral diol lipid. PMID- 10470147 TI - Induction of aromatase expression by aminoglutethimide, an aromatase inhibitor that is used to treat breast cancer in postmenopausal women. AB - Aromatase, a cytochrome P450, catalyzes three consecutive hydroxylation reactions converting C19 androgens to aromatic C18 estrogenic steroids. Aminoglutethimide (AG) is an aromatase inhibitor used to treat estrogen-dependent breast cancer. While AG is effective in inhibiting aromatase, it was found that aromatase activity in tumors of some breast cancer patients elevated after AG treatment (Miller and O'Neill, Steroids, 50: 245-252, 1987). These results may explain why some patients failed therapy after extensive AG treatment. Recently, we found that AG treatment increased aromatase activity in SK-BR-3, JAR, and HepG2 cell lines in a dose- and incubation time-dependent manner. AG induction is thought to occur at the transcriptional level because the aromatase mRNA level elevated after AG treatment in SK-BR-3 and HepG2 cells, as demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) analysis, and AG treatment did not increase aromatase activity in aromatase cDNA transfected cell lines (driven by the beta-actin promoter). Our primer-specific RT-PCR analysis revealed that in SK-BR-3 cells, AG enhanced the action of a promoter which is different from promoter I.1, I.3, or II. Furthermore, since the AG induction was found to be suppressed by SQ 22536, an adenylate cyclase inhibitor, a cAMP-dependent mechanism might be involved. Our study provides an insight as to why some patients fail therapy after extensive AG treatment. PMID- 10470148 TI - T-cell receptor V beta gene usage of human cytotoxic T-cell clones obtained from gastric cancer patients. AB - Nine T-cell clones have been established from tumor-infiltrating lymphocytes (TIL) isolated from ascitic fluid of a gastric cancer patient. Five of them retained cytotoxicity against autologous tumor cells (AuTu), and were all CD4+. Each clone had different usage of T-cell receptor (TCR) V beta gene as assessed by Southern blot analysis. Using AuTu and two allogeneic gastric cancer cell lines as targets, we selected three clones with unique cytotoxic properties. Two of these clones (Clone 1 and 2) preferentially lysed AuTu, but showed no or marginal cytotoxicity against allogeneic gastric cancer cells, and one clone (Clone 7) showed appreciable cytotoxicity against AuTu and allogeneic gastric cancer cells. In the detailed analysis of TCRV beta gene usage, Clone 1, 2, and 7 expressed V beta 13.1/D beta 1/J beta 1.5/C beta 1, V beta 3/D beta 2/J beta 2.4/C beta 2, and V beta 9/D beta 1/J beta 1.4/C beta 1, respectively, and the primary structures of the three TCRVb genes did not share any common features, neither in the sizes of their complementarity determining region 3 (CDR3) nor in their amino acid compositions. Interestingly, PBL of the same patient expressed CDR3 identical to that of Clone 2 and 7, but not that of Clone 1. CDR3 identical to that of Clone 2 and 7 were also detected in TIL of other gastric cancer patients. These results show that some AuTu-specific CTL included in TIL are circulating in peripheral blood, and that the CDR3 identical to that of the CTL is expressed extensively in TIL among different gastric cancer patients. Screening of the expression of the CDR3 in other gastric cancer patients is recommended to develop an immuno-therapy of gastric cancer based on antigenic peptide. PMID- 10470149 TI - In vitro activation of irinotecan to SN-38 by human liver and intestine. AB - BACKGROUND: Irinotecan (CPT-11) is hydrolyzed by carboxyl esterase to the active metabolite SN-38 and oral irinotecan could undergo intestinal and hepatic activation. MATERNALS AND METHODS: Irinotecan was incubated with S9 fractions of human liver and intestinal tissues and the specific activity was determined based on the formation rate of SN-38. RESULTS: Irinotecan was hydrolyzed to SN-38 by hepatic and intestinal S9 fractions with mean (+/- SD) specific activities (pmoles/min/mg) of: liver (8.57 +/- 10.4, n = 8), duodenum (5.06 +/- 3.7, n = 4), jejunum (6.44 +/- 2.8, n = 5), ileum (4.81 +/- 2.4, n = 5), colon (1.93 +/- 1.5, n = 6) and rectum (0.82, n = 1). When incubated with S9 fractions obtained from tumor tissues, there appeared to be a decrease in SN-38 formation compared to matched normal liver and colon tissues. CONCLUSION: Irinotecan undergoes conversion to its active metabolite in human intestinal S9 fractions and there is variability in the extent of SN-38 formation. The localized intestinal activation of irinotecan to SN-38 may provide a rationale for the development of oral irinotecan for gastrointestinal malignancies but could also cause mucosal damage leading to toxicity. PMID- 10470150 TI - CK20 gene expression: technical limits for the detection of circulating tumor cells. AB - The suitability of CK20 mRNA expression as a marker for the detection of minimum residual disease in patients with cancer of epithelial origin was evaluated. A sensitive nested RT-PCR assay with multiple replicates was optimised to detect a minimum number of circulating tumor cells expressing cytokeratin 20 (CK20) mRNA. Using this optimal procedure, we examined CK20 mRNA expression in ten epithelial and seven leukemic cell lines, in eight bladder tumors, in peripheral blood samples from 18 tumor patients and from 29 healthy controls and in 8 bone marrow samples from healthy donors. CK20 mRNA was found in 13 of 18 (72%) blood samples from patients with cancer of epithelial origin and in all the epithelial tumor cells tested. However, CK20 mRNA was also detected in 21 of 29 (72%) bloods, in 8 of 8 bone marrow samples from healthy donors and in 4 of 7 leukemic cell lines. These results highlight a requirement for either determination of threshold levels of CK20 normal expression or the development of quantitative techniques to distinguish between a tumor-specific CK20 gene expression and a low level background transcription of this marker. These results would also advise caution in using CK20 as a tumor specific marker in clinical investigations. PMID- 10470151 TI - Phosphorylation by DNAPK inhibits the DNA-binding function of p53/T antigen complex in vitro. AB - Interaction of p53 with Mdm2 is hindered if either protein is phosphorylated by DNA-dependent protein kinase (DNAPK), which may account for the activation of p53 in response to double-stranded DNA breaks. This finding raises the question of whether phosphorylation of p53 by DNAPK may have a general effect on its interaction with other proteins. Here we report that unlike the p53/Mdm2 complex, p53/T antigen complex remains intact following phosphorylation by DNAPK, indicating that the effect of phosphorylation upon p53 interaction is dependent on the protein partner. We have previously shown that a mouse p53/T antigen complex can bind DNA in vitro. This complex, however, was significantly reduced in its ability to bind DNA following treatment with DNAPK. This indicates that although phosphorylation did not disrupt the p53/T antigen complex, it did result in a conformational change leading to an alteration of p53' s ability to bind DNA as a protein complex. PMID- 10470152 TI - Cytotoxicity against human leukemic cell lines, and the activity on the expression of resistance genes of flavonoids from Platanus orientalis. AB - The cytotoxic activity of three flavonoids, belonging to the kaempherol series, was evaluated against 15 human leukemic cell lines. Flavonoids bearing acyl substituants, 2 and 3, were found to be the most active compounds. A further compound, 1, was examined for its ability to modulate the expression of MDR-1 and GST-pi resistance genes and compounds 2 and 3 for their effect on the uptake of [3H]-thymidine as a marker of DNA synthesis. PMID- 10470153 TI - Non-steroidal antiestrogens induce apoptosis in HL60 and MOLT3 leukemic cells; involvement of reactive oxygen radicals and protein kinase C. AB - The antitumoral activity of non-steroidal antiestrogens on promyelocytic leukemia HL60 and T lymphoblastic MOLT3 cell lines was studied. Tamoxifen and its derivatives, clomiphene and nafoxidine, caused reduction of cell viability in a dose-dependent manner. These drugs showed differences in their potency following four days incubation, with nafoxidine being the most efficient inhibitor and tamoxifen the least active. Apoptosis was induced as assessed by the DNA ladder pattern and formation of pre G0/G1 population as detected by flow cytometry analysis of DNA. The effect of these drugs was abrogated by antioxidants: alpha tocopherol was most effective in antagonizing the drugs' effect. N-acetyl L cysteine reversed mainly the decrease in cell viability caused by the drugs, but was less active on induction of apoptosis. GF109203X, a protein kinase inhibitor, attenuated apoptosis induced by clomiphene in MOLT3 cells. The results suggest that the antileukemic activity of the antiestrogens is mediated by oxidative stress and protein kinase C (PKC) activation. Triphenylethylene antiestrogens and their derivatives may be used as antileukemic drugs which kill cells by apoptosis mediated by oxidative stress and activation of PKC. PMID- 10470154 TI - Effects of intragastrically administered Pycnogenol on NNK metabolism in F344 rats. AB - NNK is a tobacco-specific nitrosamine that requires metabolic activation by cytochrome P450 enzymes. NNK may be metabolized via carbonyl reduction, N oxidation, and alpha-carbon hydroxylation. Pycnogenol is a mixture of flavonoid compounds extracted from pine tree bark and is available as a dietary supplement. We have previously shown that Pycnogenol inhibits the in vitro metabolism of NNK in lung and liver microsomes of F344 rats in a concentration-dependent manner. In this report, intragastrically administered Pycnogenol in saline affected NNK metabolism in lung microsomes differently than in liver microsomes of F344 rats. The administered Pycnogenol was inhibitory toward NNK activation in lung microsomes but not in liver microsomes suggesting that Pycnogenol may afford chemoprotection toward NNK-induced lung tumorigenesis when administered orally but not toward NNK-induced liver tumorigenesis. The effects of intragastrically administered Pycnogenol on NNK metabolism in lung and liver microsomes were similar in 6 mo and 20 mo old rats although the level of NNK metabolism was less in the 20 mo old animals. PMID- 10470155 TI - Effects of specific humoral immunity on DNA-adduct formation in Swiss mice treated with benzo[alpha]pyrene. AB - In order to study the effect of possible modulating factors on DNA-binding carcinogens, we investigated the role of specific immune response on racemic 7,t 8-dihydroxy-t-9,10-epoxy-7,8,9, 10-tetrahydro-benzo[a]pyrene ((+/-)-anti BPDE) DNA (BPDE-DNA) adduct formation. Anti BPDE Immunoglobulin G (IgG) were elicited in Swiss mice before subsequent carcinogen administration. The immunization schedule consisted of four weekly subcutaneous injections of both (+/-)-anti BPDE gelatin (BPDE-Gel) and DNA (BPDE-DNA) conjugate, followed by a final immunogen injection 14 days later. The immunization procedure resulted in the production of specific anti-BPDE antibodies in all treated animals. One week after the end of the immunization procedure, both groups of immunized and non immunized mice were treated with different doses of B[a]P (25-50-100-200 mg B[a]P/Kg body weight) by intraperitoneal injection. Seven days after treatment, the mice were sacrificed. Adduct levels were detected by competitive ELISA by using optimal conditions established in our laboratory and highly specific and sensitive IgG anti BPDE-DNA induced in rabbit. The determination of DNA adducts in liver revealed significantly lower B[a]P adduct levels in liver of immunized mice with respect to non-immunized animals. This result confirms those obtained for 2 acetylaminofluorene (2-AAF) in a previous work: the specific humoral immunity elicited by repeated carcinogen exposure may be able to modulate the genotoxic effect induced by subsequent carcinogen administration. PMID- 10470156 TI - Loss of function of the tissue specific transcription factor HNF1 alpha in renal cell carcinoma and clinical prognosis. AB - BACKGROUND: Human renal cell carcinogenesis is associated with loss of expression of tissue-specific genes and loss of function of tissue-specific transcription factors such as HNF(hepatic nuclear factor)1 alpha. MATERIALS AND METHODS: In this study HNF1 alpha DNA-binding activities and protein amounts were determined by gel retardation assay and Western blot analysis, respectively, in 42 non metastasized renal cell carcinomas and paired normal tissues. RESULTS: 36 tumors out of 42 (86%) showed diminished binding activity of HNF1 alpha. In most cases (26 out of 42) this appeared to be due to decreased amounts of HNF1 alpha protein, but 10 tumors contained equal or even higher amounts of HNF1 alpha, in spite of reduced binding to DNA. Only 6 tumors out of 42 had unaltered HNF1 alpha binding activity. A clinical follow-up was obtained for 40 patients. Over an average follow-up period of 39 months no significant differences in the survival rate were observed between patients having lost or retained HNF1 alpha function. However, since most of the patients with retained function are still alive, long term follow-up might be warranted. CONCLUSIONS: The very high incidence of loss of HNF1 alpha function indicates the important biological role of this change in renal cell carcinoma. PMID- 10470157 TI - Lack of prognostic significance of ploidy and S-phase measurements in advanced ovarian cancer. AB - DNA ploidy and S-phase measurements were made in fresh tumour samples obtained from 27 patients with either untreated or previously treated ovarian cancer. In addition, an assessment of in vitro chemosensitivity to post-operative chemotherapy was carried out. Whilst we found that patients with aneuploid tumours tended to survive longer (median survival 441 days versus 268 days for diploid tumours; p = 0.308) and this was associated with clinical response (p = 0.137) this was not statistically significant. Tumours with a high S-phase fraction (i.e. > 9% median) showed a median survival of 362 days compared with 484 days for those with a low S-phase fraction (< 9%; p = 0.79). Whilst a high level of predictability for chemosensitive patients (81%) was achieved, chemosensitivity per se was not related to the respective tumour DNA ploidy or S phase fraction. In addition, when considering disease stage i.e. FIGO stage III or IV, or extent of residual disease between diploid and aneuploid groups, there were no statistically significant differences. However, there was a tendency for stage III tumours to be aneuploid (p = 0.189). Aneuploid tumours also showed the highest S-phase fractions (p = 0.03), indicative of a high proliferation rate. In conclusion, we have shown that DNA content and ploidy status do not appear to be major prognostic indicators for the management of ovarian cancer. PMID- 10470158 TI - Synthesis and antiproliferative activity of N-acylaspartic acid dimethyl esters. AB - Farnesyl residues are found as a lipophilic modification of a number of important proteins. In addition, synthetic farnesyl derivatives display a range of biological effects. We have prepared a series of N-acylaspartates as structural analogs of farnesylpyrophosphate in which the farnesyl residue has been replaced by a number of different aliphatic and aromatic carboxylic acids and the aspartate is used as a pyrophosphate surrogate. The corresponding dimethyl esters of these aspartates were assayed against different tumor cell lines. Several N acylaspartic acid dimethyl esters carrying an aromatic acyl residue displayed a selective antiproliferative effect against THP-1 cells with GI50 values ranging from 7.6 to 1.3 microM. PMID- 10470159 TI - Sodium phenylacetate (NaPa) induces modifications of the proliferation, the adhesion and the cell cycle of tumoral epithelial breast cells. AB - Sodium phenylacetate (NaPa), a physiological product of phenylalanine metabolism, present in micromolar concentrations in human plasma, has been shown to induce in vivo and in vitro cytostatic antiproliferative effects at millimolar concentrations. Cadherin molecules are powerful invasion suppressor molecules and the reduction of E-cadherin expression plays an important role in the invasion and metastasis of human breast cancer. In this study, we demonstrated, on one hand, that NaPa stimulated aggregation by increasing the expression of E-cadherin at the surface of breast cancer MCF-7ras cells transformed by Ha-ras oncogene and inhibited its expression in MCF-7 cells. We demonstrated that NaPa increased the formation of MCF-7ras cell aggregates and did not alter the formation of MCF-7 cell aggregates. By Northern blot, we demonstrated that the E-cadherin expression was not regulated at the transcriptional level. On the other hand, we analyzed the cell cycle of these 2 cell lines after NaPa treatment and showed that NaPa induced arrest at the G1/S phase in both MCF-7 and MCF-7ras cells. bFGF increased the growth of MCF-7 cells, but inhibited MCF-7ras cell proliferation. NaPa treatment suppressed the stimulation of MCF-7 cell proliferation and increased MCF-7ras cell growth inhibition. We have demonstrated a new target of NaPa action in blocking the cell cycle of tumor cells in G0/G1. We suggest that the anti proliferative effect of NaPa associated to the restoration of the cadherin function in human mammary carcinoma cells indicates that NaPa could be a novel therapeutic agent in breast cancer. PMID- 10470160 TI - Synthesis and antiproliferative activity of 2-triazenothiophenes. AB - A series of 2-triazenothiophene derivatives was prepared and tested to evaluate their biological activity. Two compounds inhibited the proliferation of leukemia, lymphoma and solid tumor-derived cell lines at micromolar concentrations, whereas none of the compounds were active against HIV-1. Compound 3c inhibited DNA, RNA and protein synthesis, and was also effective against KB cells resistant to etoposide and vincristine. The compounds were inactive against fungi and bacteria. PMID- 10470162 TI - Immunohistochemical detection of HSP60-expression in human ovarian cancer. Correlation with survival in a series of 247 patients. AB - PURPOSE: In a previous pilot study, HSP60 expression at the transcriptional (mRNA) level was shown to be a negative prognostic factor in ovarian cancer. The aim of this study was to determine HSP60-expression by means of immunohistochemistry and to correlate the results with survival in a large series of ovarian carcinoma patients with a closed follow-up. MATERIALS AND METHODS: Slides from routinely processed, paraffin-embedded tumor blocks belonging to 247 patients with epithelial ovarian carcinoma were studied for the overexpression of HSP60 using the Lk2 monoclonal antibody and the strepatvidin-biotin-peroxidase technique. HSP60-expression was correlated with overall survival by means of life table analysis and the Kaplan-Meier method. RESULTS: 47 tumors (19%) expressed HSP60. Of them, 12/29 (41.4% positivity) were stage I tumors, whereas only 35 out of the remaining 218 tumors in more advanced surgical stage (16.1%) showed HSP60 staining. This difference was statistically significant (Fisher s exact test; p = 0.004). Even when stratifying stage for stage, the difference between groups still remained statistically significant (Chi square test; p = 0.0095). The survival curve analysis showed a significant difference in favor of those tumors expressing HSP60 (median survival 28 vs. 37 months; log-rank test, p = 0.02). CONCLUSION: Immunohistochemical detection of HSP60-expression in human epithelial ovarian carcinoma is significantly more frequent in tumors from patients with initial stages of the disease. Therefore, HSP60-expression determined by this method is associated with a significantly better prognosis. PMID- 10470161 TI - Immunohistochemical localization of metallothionein in human breast cancer in comparison with cathepsin D, stromelysin-1, CD44, extracellular matrix components, P53, Rb, C-erbB-2, EGFR, steroid receptor content and proliferation. AB - Metallothionein (MT) is a low molecular weight, cysteine-rich, zinc-binding protein that may have a function in cellular repair processes, growth and differentiation. Using a monoclonal antibody (E9) to metallothionein, we investigated the immunohistochemical expression of MT in routinely fixed and paraffin-embedded tissue from 98 cases of female breast carcinomas. The MT expression was studied in comparison with the expression of the basement membrane (BM) antigens (type IV collagen, laminin), fibronectin, cathepsin D, adhesion molecule CD44, p53 protein, the pRb, c-erbB-2 oncoprotein, EGFR, stromelysin-1, proliferation indices (Ki-67, PCNA), steroid receptor content as well as with other conventional clinicopathological parameters of breast cancer. Strong MT expression was observed in the majority of tumour cells in 18.4% of tumours, focal MT positivity in 13.3% and almost complete lack of MT expression in 68.4% of cases (mean value 33.36 +/- 26.36). The MT expression in carcinoma cells was strongly associated with the DCIS component of the tumour (p < 0.0001). High values of MT were correlated with low steroid receptor status (p = 0.08 for ER receptor and p = 0.019 for PgR receptor content). MT positive cases were correlated with stromelysin-1 expression (p = 0.059) and cathepsin D (p = 0.058). These findings suggest that MT expression is characteristic of the early phase of breast carcinogenesis, possibly regulated by hormones, and could be a new potential prognostic marker in breast cancer. PMID- 10470164 TI - Tumors of the gastrointestinal tract and the pancreas: histopathology, staging and prognosis. AB - At present, the pathologist is an important member of the treatment team who provides information and consultation for selecting treatment, estimating prognosis and evaluating outcome. The pathologist's functions include pretreatment microscopic diagnosis, intraoperative consultation and histopathologic examination of tumor resection specimens. Tumor classification follows international rules published by the UICC and the WHO. The strongest predictors of outcome are the residual tumor (R) classification and the TNM/pTNM classification of anatomic extent before treatment. The new 5th edition of TNM (1997) contains some changes in the classification of gastric, colorectal and exocrine pancreatic carcinoma. There are some pathological features which independently influence prognosis in addition to R and TNM. For the most biological and molecular markers the prognostic significance remains to be proven. In future, the main objective of prognostic factor research will be the development of prognostic systems. PMID- 10470163 TI - Immunohistochemical evaluation of cathepsin D expression in colorectal tumours: a correlation with extracellular matrix components, p53, pRb, bcl-2, c-erbB-2, EGFR and proliferation indices. AB - The immunohistochemical Cathepsin D (CD) expression of tumour and stromal cells was investigated in a series of 93 human colorectal adenocarcinomas and 22 adenomas with the intention to evaluate its prognostic significance and its contribution in the metastatic potential of colorectal cancer. CD expression was correlated with the expression of extracellular matrix components (collagen type IV, laminin and fibronectin), p53 protein, pRb, bcl-2, c-erbB-2, EGFR, proliferation indices (Ki-67, PCNA) as well as with other conventional clinicopathological features. CD expression (> 10% of positive tumour cells) was observed in 60.2% of carcinomas and in 72.7% of adenomas. Stromal CD expression was detected in all cases. A statistically significant positive correlation between neoplastic cells CD and stromal cells CD (SCCD) was observed in both carcinomas and adenomas. Cancer cells CD (CCCD) was positively correlated with collagen type IV and pRb expression as well as with PCNA score. In carcinomas, SCCD expression was statistically correlated with p53 protein and pRb expression and a trend for correlation with PCNA score was found. These data suggest that Cathepsin D of cancer and stromal cells, especially in combination with other markers, may provide more information about the biological behaviour of colorectal cancer. PMID- 10470165 TI - TNM staging, histopathological grading, and tumor-associated antigens in patients with a history of mucoepidermoid carcinoma of the salivary glands. AB - BACKGROUND: The aim of this study was to evaluate the TNM categories and histopathological grading as prognostic factors in mucoepidermoid carcinoma (MEC). In addition, the study was designed to provide baseline data on levels of tumor-associated antigens (TAA) in sera of MEC patients. MATERIALS AND METHODS: Fifty-nine patients with MEC of the salivary glands were evaluated. RESULTS: Reclassified TNM stage, at the time of initial diagnosis, varied considerably. In disease-free patients, none of the tested sera were TAA elevated above the cut off levels. Our patients who died of tumor metastasis had all been classified as stage III or IV at the time of initial diagnosis. Distant metastases are rarely found even decades after surgical therapy. CONCLUSIONS: Long-term follow-up is recommended for patients with MEC of the salivary glands. TAA are not elevated in disease-free patients. The value of TAA in the monitoring of MEC patients remains to be evaluated in further studies. PMID- 10470166 TI - Immunohistochemical detection of cytokeratins in the irradiated rat mandibular gland. AB - BACKGROUND/MATERIAL AND METHODS: The present study is a prerequisite for the exploration of the cytokeratin (CK) profile in rat salivary gland tumors. In 59 rat mandibular glands we investigated the distribution of CK by immunohistochemical analysis. The animals differed in age and pretreatment status (irradiation versus no irradiation). RESULTS: The monoclonal antibodies (CKE3, Ks13.1, NCL5D3, K8.12; against CK 17, CK 13, CK 8 and CK 13/15/16, respectively) identified different epithelial structures in rat salivary gland tissue, including intercalated duct cells (ICD), striated duct cells (SD), granular convoluted tubules (GTC), excretory duct cells (ECD) and myoepithelial cells (MC). As typical results, CKE3 usually stained the ICD, SD and ECD moderately to strongly and stained the GTC slightly. K8.12 staining was restricted to ECD and MC. Differences in immunoreactivity were seen between irradiated and non irradiated glands, predominantly with stronger staining in the irradiated group. CONCLUSIONS: Previous radiation has to be considered when interpreting immunohistochemical stainings of salivary gland tissue, especially in tumor differentiation studies following irradiation. PMID- 10470167 TI - NuMA: evaluation of a new biomarker for the detection of low stage colorectal cancer. AB - An enzyme immunoassay for NuMA was evaluated in a retrospective clinical study for its potential utility in the detection of colorectal cancer. The concentrations of NuMA and CEA (Abbott IMx) were measured in sera from 86 patients (presurgical) with colorectal cancer, 72 subjects with benign gastrointestinal diseases, 80 subjects with risk factors for colorectal cancer, and 141 age-matched healthy subjects. Reference values for NuMA and CEA were calculated by two methods: 95% cumulative distribution and ROC analyses versus healthy subjects. By the first method, NuMA and CEA both had approximately 20% sensitivity for colorectal cancer. By the second method (which generated lower reference values), NuMA was more sensitive than CEA for colorectal cancer. This improved sensitivity was most evident in Dukes B subjects. By either analysis method, NuMA was more sensitive than CEA for subjects at risk for developing colorectal cancer, whereas CEA was more specific for benign gastrointestinal diseases. PMID- 10470169 TI - Value of human chorionic gonadotropin compared to CEA in discriminating benign from malignant effusions. AB - Human chorionic gonadotropin (HCG) is expressed in germ cell tumors and urothelial, breast, lung and colon cancers. The aim of the study was to investigate if the determination of HCG in comparison with CEA is able to discriminate between malignant and benign effusions. Effusion and partially serum samples of 61 patients with benign (g.i., heart/kidney isnuff.) and 116 patients with malignant diseases (g.i., gynec., lung, misc., CUP) were investigated. HCG was specifically determined by an IRMA using 2 monoclonal antibodies, CEA by a conventional double Ab RIA. Cytological staining was preformed using the Pappenheim-method on cytospin preparations. Significant differences (p < 0.001) were found for HCG between benign and malignant ascitic effusions with the best discrimination at 5 IU/l (ROC) and an overall sensitivity of 31.3% (spec. vs benign eff. 93.4%) increasing in subgroups from hematol. (5.8%) < misc. (31.3%) < gynec. (32.1%) < g.i. (36%) < lung (38.1%) to CUP (50%). CEA also showed significant differences between benign and malignant total and ascitic effusions, and weaker for the pleural subgroup (cutoff 9 ng/ml) with a total sensitivity of 44.6% (sp = 100%) increasing from misc. (30.8%) < lung (47.1%) < CUP (50%) < gynec. (60%) < g.i. (60.9%). Comparative cytology and TM determinations increased the positiverate of cytology (45.2%) to 58.3% for either cytology or HCG positive cases, or to 61.6% for either cytology or CEA positive cases. For the combined determination of cytologoy and HCG and CEA, the overall TM positive rate for 33 cytology-pos. cases was 78.8%, but in 40 cytology-negative cases 37.5% for TM positive cases. In conclusion HCG is useful in ascitic > pleural effusions with high specificity (90% at 5 IU/l) but low sensitivity of 31% increasing in g.i., lung and gynecologic cases, CEA a more general TM with higher sensitivity of 45% increasing in g.i., gynecologic and lung cases (sp. 100% at 9 ng/ml) both adding significantly to cytology-negative effusions. PMID- 10470168 TI - Nuclear mitotic apparatus protein (NuMA) in benign and malignant diseases. AB - Nuclear mitotic apparatus protein (NuMA) is a 239 kDa internal nuclear matrix protein described to be elevated in cancer patients, especially in colorectal carcinoma and early colorectal cancers. We tested the significance of NuMA as tumour marker in colorectal cancer and also the sensitivity/specificity profile in general. Therefore, we investigated in a retrospective clinical study 507 sera from patients suffering from solid tumours, with the main emphasis on colorectal carcinoma, and 418 sera from patients with benign diseases and healthy individuals. Testing was done with a double monoclonal enzyme immunoassay detecting head and rod domain of NuMA and results were compared to the established tumour associated antigens. Based on a specificity of 95% versus the benign reference group of gastrointestinal diseases, we found--at the time of primary diagnosis--a sensitivity for colorectal cancer of 8% for NuMA, 36% for CEA and 17% for CA 19-9. Regarding T-stages of colorectal cancer no marker detected T1 when regarding 95% specificity-cut-off value but NuMA showed little more sensitivity when based on a 95% specificity cut off value versus healthy. This could not be shown in Dukes' stages. Regarding all other solid tumours tested--all based on a specificity of 95% for the corresponding benign reference groups--no advantage of NuMA in sensitivity for all other solid tumours investigated was found. No additional sensitivity could be observed. Based on our results, the NuMA-assay in its present form has no clinical relevance. PMID- 10470170 TI - Radioimmunotherapy of colorectal cancer in small volume disease and in an adjuvant setting: preclinical evaluation in comparison to equitoxic chemotherapy and initial results of an ongoing phase-I/II clinical trial. AB - The 5-year survival of colorectal cancer patients with distant metastases is below 30%, despite the development and use of a variety of chemotherapeutic regimens. Therefore, new therapeutic strategies are warranted. Whereas radioimmunotherapy (RIT) has shown disappointing results in bulky disease, it may be a promising therapeutic alternative in limited and small volume disease. The aim of this study was, therefore, to compare, in a preclinical study, the therapeutic efficacy of RIT in colorectal cancer to equitoxic chemotherapy, as well as to evaluate, in a pilot clinical trial, its efficacy in small volume disease. Nude mice, bearing subcutaneous or metastatic human colon cancer xenografts, were injected either with the unlabeled or 131I-labeled monoclonal antibodies (MAbs), CO17-1A (which is a murine IgG2a directed against a 41-kD membrane glycoprotein) or F023C5 (which is an anti-CEA MAb of murine IgG1 subtype), or were administered 5-fluorouracil/folinic acid (5-FU/LV) at equitoxic doses. In a pilot clinical study, 10 colorectal cancer patients with small volume metastatic disease (all lesions < or = 3 cm) have been entered so far in an ongoing mCi/m2-based dose escalation study with the 131I-labeled F023C5. In the animals, the maximum tolerated activities (MTD) of 131I-labeled CO17-1A and F023C5 were 300 microCi and 600 microCi, respectively, corresponding to blood doses of approximately 15 Gy each. Accordingly, myelotoxicity was dose-limiting. The MTD in the chemotherapy group was 0.6 mg 5-FU/1.8 mg LV, given as intravenous bolus 1 h apart for 5 subsequent days. Whereas no significant therapeutic effects were seen with both unlabeled MAbs or 5-FU/LV chemotherapy, tumor growth was retarded significantly with both radiolabeled antibodies. In the metastatic model, chemotherapy prolonged life for only a few weeks, whereas RIT led to cures in 35-55% of the animals. As was the case in the animals, myelotoxicity seems to be dose-limiting in patients as well. Encouraging anti-tumor effects were observed, lasting for up to more than 12 months. These data suggest that radioimmunotherapy may be a viable therapeutic option in colorectal cancer patients with limited disease. Myelotoxicity is the only dose-limiting organ toxicity. Although most patients were treated below the MTD, anti-tumor effects are encouraging. Further studies are ongoing. PMID- 10470171 TI - CA19-9: a pedictor of response in pancreatic cancer treated with gemcitabine and cisplatin. AB - The question was asked whether kinetics of CA19-9 would serve as a predictor of chemotherapeutic outcome in advanced pancreatic cancer treated with gemcitabine and cisplatin. Twenty one patients, 5 females and 16 males (median age 56 yrs, range 36-71 yrs) suffering from adenocarcinoma of the exocrine pancreas were analysed. Chemotherapy was applied for a median of 6 courses (range 2-21). Four patients achieved a complete remission, four a partial remission (OR = 38%), while stable disease was documented in 8 and progressive disease in 5 patients. Among 4 CR patients, all demonstrated a significant decline of CA 19-9 levels during the initial three treatment courses with apparent half-lifes of 15, 18, 24, and 33 days. At a cut-off level of 37 U/mL, all CR patients reached normal values in the course of treatment. All patients achieving PR showed a decrease of CA 19-9 values at apparent half-lifes of 9, 16, 88 and 89 days. Among patients with stable disease, CA19-9 transiently decreased in 7/8 patients and remained stable in 1 patient. However, values increased later in all patients after a median of 3 treatment courses (range 2-9). In patients with disease progression, CA 19-9 initially increased in 4/5 patients, while a further patient did so only beyound 100 days of treatment. In conclusion, kinetics of CA19-9 serum concentration may serve as an early indicator of response to gemcitabine/cisplatin chemotherapy in advanced pancreatic cancer. PMID- 10470172 TI - 18-FDG positron emission tomography of the pancreas: diagnostic benefit in the follow-up of pancreatic carcinoma. AB - The aim of this prospective study was to determine the role of FDG-PET in the follow up care of patients with pancreatic carcinoma in comparison with sonography, computed tomography and tumour marker levels. 19 patients were investigated, 9 patients with non-resectable tumour disease and 10 patients after resection of primary tumor. We found corresponding results in 15/19 patients, but in 2 patients additional metastases were detected. In another 5 patients of this group, PET results decisively influenced the further therapeutic procedure. In 2/19 patients only PET showed viable tumour tissue, in spite of negative findings in CT and sonography. In 2/19 patients we found false negative results with PET. One patient had increased blood glucose levels at the time of investigation, the other one was investigated shortly after locoregional chemotherapy. Compared to CA 19-9, PET investigation may also result in positive findings, in spite of non elevated CA 19-9 serum levels. CONCLUSION: PET has been shown as an effective tool also in the follow up care of patients with pancreatic cancer. In more than half of the patients PET offered important additional information to clinicians, resulting in a change of the therapeutic procedure. Especially before locoregional chemotherapy or in case of increased tumor marker levels without morphological substrate in US/CT PET with FDG seems to represent a valuable investigation. PMID- 10470173 TI - CEA, TPS, CA 19-9 and CA 72-4 and the fecal occult blood test in the preoperative diagnosis and follow-up after resective surgery of colorectal cancer. AB - In a prospective clinical study we examined the diagnostic procedures used in the preoperative diagnosis of colorectal cancer patients (n = 176) and the value of supplementation of standard diagnostic methods (clinical investigation, colonoscopy, barium enema, ultrasound, computer tomography) with a test for occult fecal blood (FOBT) and an expanded tumor marker panel (CA 19-9, TPS and CA 72-4 in addition to CEA) in the postoperative follow-up (n = 116, mean follow-up 21 months). Preoperative diagnosis based on colonoscopy/barium enema, followed by histology and the imaging methods, in most cases US and/or CT. Patients with postoperative stage Dukes D and after palliative surgery were excluded from the follow-up study (n = 43). The patients were seen every three months (clinical investigation, US, CT, tumor markers, FOBT) within the first two postoperative years and every half a year thereafter. 83 of the 116 patients (16%) developed a recurrent disease and 5 of them could be reoperated with curative intention. In addition to the 19 patients 14 simultaneously admitted patients with recurrence of colorectal cancer (total n = 33) were studied. The results of our study analyzing the sensitivity and specificity of colonoscopy, tumor, markers and the FOBT in the preoperative and postoperative phases as well as in the diagnosis of recurrent disease of colorectal cancer confirms the view that FOBT and tumor markers cannot replace endoscopic and imaging methods. However they support the concept, that diagnosis and follow-up of colorectal cancer should be based on a combination of clinical investigation and imaging methods (US, CT etc. and endoscopic and/or x-ray examination) with supplementation by FOBT and determination of tumor markers, mainly CEA. In the case a patient is asking for prognostic and recurrence information as early and as valid as possible we currently recommend the following procedure for the first two years after surgery: every three months the determination of tumor markers, FOBT as well as ultrasound of the upper abdomen and CT of the lower abdomen and every half year a total colonoscopy. The question of whether this program will also have a therapeutical relevance depends on several factors like the personal experience and concepts of the consulting surgeons and oncologists and also on the tasks and duties which are considered worthy by the patient for his further life if confronted with the diagnosis of recurrent colorectal cancer. PMID- 10470174 TI - MR-cholangiopancreaticography (MRCP) and MR-angiography: morphologic changes with magnetic resonance imaging. AB - Since 1997 we have prospectively tried to assess/confirm the diagnostic value of MR-Cholangiopancreaticography (MRCP) and MR-Angiography in patients suffering from pancreatic carcinoma. Till today we have studied 116 adult patients with two 1,5 Tesla MRT scanners using a body phased-array coil: 27 patients with benign diseases of the pancreas, 58 with carcinoma of the pancreas, 15 with no disorder of the pancreatobiliary system, 14 with hepatic abnormalities and 2 with other tumors. MR examinations included routinely T1-weighted (TSE, SE), T1-weighted fat suppressed (TSE, SE), T2 weighted fast turbo spin-echo (UTSE, Haste), T2-weighted fat-suppressed fast turbo spin-echo (SPIR), 2D-cholangiopancreaticography (images were obtained with breath-held) or 3D-cholangiopancreaticography (images were obtained in coronal and axial plane with respiratory triggering). In cases with pancreatic cancer we added MR-angiography. MR images were retrospectively compared with CT, ERCP amd sonography data. Summarizing the results confirm that MRCP is a safe and non-invasive technique for studying pancreatic and biliary diseases which yields information in many cases complemontory to ERCP and PTC. MRCP images can also be used as a guide for subsequent interventional procedures. MRCP will, therefore, allow the restriction of ERCP and PTC to more therapeutical indications and cases offering special problems. Considering staging and follow up of pancreatic cancer, MRCP cross-sectional images and MR-angiography should be performed, allowing the visualization of the extraductal anatomy/pathology. These techniques may provide the clinicians with complementary information compared to other conventional imaging methods like US and CT. Clinicians engaged in pancreatic oncology, therefore, should have MRCP, MR-angiography and MR imaging available in addition to the conventional diagnostic tools. PMID- 10470175 TI - Locoregional/systemic chemotherapy of locally advanced/metastasized pancreatic cancer with a combination of mitomycin-C and gemcitabine and simultaneous follow up by imaging methods and tumor markers. AB - Stimulated by surprising results in the first 7 patients treated with intraarterial/systemic chemotherapy with a combination of Mitomycin-C + Gemcitabine we now report on the data of 28 pancreatic cancer patients in comparison to 15 patients treated with gemcitabine alone(1000 mg/m2). Tumor response was evaluated on the basis of imaging methods, tumor markers and life quality parameters like body weight, pains and Karnofsky index etc. Tumor markers were monthly determined, imaging methods every 1-3 months. The locoregional/systemic approach included 3 week cycles with i.a. application of mitomycin-C + gemcitabine on day 1 and i.v. application of gemcitabine on days 8 and 15. The alltogether 125 cycles (mean 4 cycles per patient) resulted in 43% PR (n = 12), 1 CR (3%), 255 MR, 11% SD and 18% PD in the imaging methods and 20% CR, 60% PR, 4% MR and 12% SD in the course of the relevant tumor markers. Progression free survival amounted to a median of 7.5 (6) months defined by imaging methods (tumor markers). Second line treatments following new progress after effective locoregional approach with gemcitabine or a combination of gemcitabine + oxaliplatin did not result in a new tumor regression. However, third line therapy trials in 3 patients with high dose 5-FU or CPT 11 induced new antitumoral efficacy (survival of these tumors > 15, > 17 and > 28 months, n = 2 M1, n = 1 T3M0). About 75% of patients reported on a relevant benefit of life quality parameters. Side effects are on principle comparable to those of gemcitabine monotherapy, except for a tendency to a higher rate of pulmonary complications and 1 HUS observed. Even if not compared in a randomized study locoregional/systemic combined treatment modality seems to result in a higher rate of abjective tumor response defined by imaging methods as well as tumormarkers. Comparing tumor marker and imaging response to therapy CA 19-9 often showed a more rapid and subtle answer to therapy and an earlier new increase suggesting tumor markers as an essential part in the follow-up of these patients in order to optimize the patient's palliative treatment. Our results should stimulate the clinicians to rediscuss the chemosensitivity of exocrine pancreatic cancer and to perform prospective randomized studies focusing on combined gemcitabine approaches. PMID- 10470176 TI - Pancreatic cancer associated ascites-derived CTL recognize a nine-amino-acid peptide GP2 derived from HER2/neu. AB - BACKGROUND: The proto-oncogene HER2/neu encodes a 185 kDa transmembrane protein with extensive homology to the epidermal growth factor receptor. It is overexpressed in several human cancers of epithelial origin, such as pancreatic cancer. Previously, we demonstrated that CTL derived from breast, ovarian, and non-small cell lung cancer recognized a peptide derived from HER2/neu. The aim of this study was to evaluate whether this HLA-A2-binding peptide is a TAA in pancreatic cancer and if pancreatic cancer associated T-lymphocytes (TAL) are useful to generate tumor- and peptide-specific CTL. MATERIALS AND METHODS: TAL from malignant ascites of a HLA-A2+ pancreatic cancer patient whose tumor overexpressed HER2/neu were stimulated on solid-phase anti-CD3 and cultured in low-dose IL-2. Using repetitive autologous tumor cell stimulation, CTL were generated. RESULTS: CTL recognized autologous and allogeneic HER2/neu+ tumor cells in an HLA-A2 restricted fashion significantly. Furthermore, all CTL recognized p654-662 (GP2) derived from HER2/neu, but not the control peptide. CONCLUSIONS: These results demonstrate that this HER2/neu derived peptide is a TAA in pancreatic carcinoma. The identification of the HER2/neu derived peptide GP2 as a TAA in pancreatic cancer provides an opportunity for the design of novel immunotherapy and vaccine strategies. The possibility of generating peptide specific CTL from malignant ascites enables future studies to identify more antigens in this disease. PMID- 10470177 TI - CA 242 in comparison with established tumour markers in colorectal, pancreatic and lung cancer. AB - In a prospective study (N = 566) we investigated the tumour associated carbohydrate-protein CA 242, focusing on the question whether CA 242 (CaNAG, Sweden) expression in carcinoma patients is distinctly higher than in benign disorders, especially when compared to CA 19-9 (EIA Roche Germany). A second point of interest was if CA 242 is expressed to a higher extent in early stages of colorectal cancer than CEA (MEIA Abbott, USA) and CA 19-9 are, and third its behavior in pancreatic and lung cancer. We found CA 242 values comparable in healthy individuals and benign gastrointestinal disorders, thus CA 19-9 remains the marker of first choice for pancreatic cancer and CEA for colorectal cancer. CA 242 shows no advantage in lung cancer as compared to the established markers (CEA, CYFRA 21-1 (EIA Roche Germany) and NSE (EIA Hoffmann LaRoche, Switzerland) and no clearly higher expression in early colorectal cancer. Overall, the combination of CEA and CA 242 shows the best sensitivity in colorectal cancer. PMID- 10470178 TI - Analysis of K-ras mutations in pancreatic tissue after fine needle aspirates. AB - The majority of pancreatic carcinomas contain a mutation at codon 12 of the K-ras oncogene. We have analysed 87 samples from 76 patients who underwent surgery because of different pancreatic diseases to evaluate whether the detection of K ras mutations may be helpful to discriminate between chronic inflammation and neoplastic growth. Mutation analysis was performed using a semi-nested PCR followed by a selective restriction enzyme digestion. The correlation of clinical follow ups with the results of the molecular analysis was performed from 47 patients. K-ras mutations were detected in 50% of adenocarcinomas and no point mutation was found in normal pancreatic tissue and in tumor tissue from entities other than pancreas. Otherwise, K-ras mutations were detected in tissue samples from two patients with chronic pancreatitis, and one patient was found to have an adenocarcinoma after additional clinical investigation. Further studies especially follow ups will be helpful to get a better insight into the pathogenesis of pancreatic tumors and may be useful as an early diagnostic test. PMID- 10470179 TI - Trofosfamide in patients with pancreatic cancer. AB - BACKGROUND: In a previous phase II trial of ifosfamide (IFO) several patients with pancreatic cancer responded to therapy (1). Trofosfamide (TROFO) is an orally taken oxazaphosphorine prodrug. Metabolic products of TROFO include ifosfamide (IFO) and cyclophosphamide (CYCLO). In clinical pilot studies with TROFO against refractory malignancies mild toxicity was reported. PATIENTS AND METHODS: Since virtually all patients with advanced pancreatic cancer are symtomatic, we designed and conducted a prospective phase II trial to evaluate the activity of single agent therapy with TROFO given orally continuously (three times 50 mg daily), and to assess the toxicity, duration of remission, and overall survival of the treated patients. RESULTS: 16 patients were enrolled onto the study. In one patient a PR has been observed; the mayor response rate was 6%. The mayor toxicity was bone marrow toxicity with granulocytopenia (WHO grade II) and anemia (WHO grade II) in virtually all patients. CONCLUSION: Single agent therapy with TROFO in patients with pancreatic cancer is safe and well tolerated; however, it shows low clinical activity. PMID- 10470180 TI - Use of serum PIVKA-II (DCP) determination for differentiation between benign and malignant liver diseases. AB - BACKGROUND: The concentration of Des-gamma-carboxy-prothrombin (DCP) or PIVKA-II has been described to be increased in patients with hepatocellular cancer, along with its elevation in vitamin K deficient states by warfarin or dicoumarol treatment. The aim of the study was to investigate its clinical value in HCC in comparison with alpha-fetoprotein. PATIENTS AND METHODS: Measurements were performed in duplicate in serum of 87 patients with benign (acute/chronic hepatitis B/C/autoimmune, liver cirrhosis B/C/alcoholic) and 154 patients with highly probable (CT, MRT imaging) or histologically proven HCC. Two commercial or research ELISA tests (1: Eitest MonoP-II, Eisai, Tokyo, Japan; 2: Asserachrom PIVKA-II, Stago, France) using murine monoclonal anti-PIVKA-II antibodies were used comparatively and compared with a laboratory-developed conventional AFP-RIA. RESULTS: By ROC analysis, a highly significant discrimination (p < 0.0001) was found at cutoffs of 0.09 AU/ml (Eisai) or 0.8 ng/ml (Stago) at a specificity of about 90% (Eisai: s = 78.6%, ppv = 0.92, npv = 0.70; Stago: s = 77.9%, ppv = 0.93, npv = 0.70) compared with the AFP-test at a cutoff of 45 ng/ml (sp = 91%, s = 58.4%, ppv = 0.92, npv = 0.55). A higher significant correlation was seen between both DCP tests in malignant (rS = 0.89, p < 0.0001) than benign groups (rS = 0.41, p < 0.001) and a lower correlation between the AFP and Eisai (rS = 0.27/0.36, p < 0.01) and Stago test for the malignant group (0.16; p < 0.05). CONCLUSION: DCP determination in serum/plasma adds significantly in the discrimination between benign and malignant liver diseases. PMID- 10470181 TI - Differential tumor markers and hepatitis markers profile in liver tumors. AB - The differential diagnostic utility of AFP, CEA, CA19.9 and TPA was evaluated in liver tumors. They were determined in the sera of 61 patients with primary hepatocellular carcinoma (HCC), 18 with secondary liver metastasis, 61 of benign liver cirrhosis in comparison to 20 normal healthy control subjects. The association of either HBV or HCV infection and HCC was also studied through the assay of HbSAg, HbSAb, and HCV-Ab. The optimal cut-off values were determined using the diagnostic accuracy measurements and the receiver operating characteristic (ROC) curves. AFP at an optimal cut-off value of 100 ng/ml and TPA at 160 U/L showed the highest sensitivity and specificity in detecting liver metastasis (100% and 87% for AFP; 100% and 54% for TPA respectively). The obtained data indicated that the combined assay of AFP and TPA resulted in a better discrimination of HCC among patients with hepatic focal lesions. HCV-Ab was detected in a higher ratio of HCC patients (83.6%) compared to HbsAg (68.9%), and both were detected in (34%) of HCC patients. This high incidence of HCV-Ab may suggest the implication of HCV in the molecular events leading to hepatic carcinogenesis. PMID- 10470182 TI - Determination of the oncogenes p53 and C-erb B2 in the tumour cytosols of advanced hepatocellular carcinoma (HCC) and correlation to survival time. AB - Mutations in the p53 gene are the most frequently reported somatic gene alteration in human cancer. Overexpression of the oncogene c-erb B2 takes place also in many human carcinomas, esp. in cancers of the breast, colon and ovary. High values of both oncogenes correlate with poor prognosis. We analysed the concentration of p53 in tumor cytosols of 34 primary advanced HCC and with the clinical outcome and survival time. p53 was analysed in the tumor cytosol by luminometric immunoassay (LIA) using monoclonal antibodies detecting wild type and mutant p53 protein. C-erb B2 protein is analysed by a monoclonal ELISA. p53 values vary from 0 to 8.1 ng/mg protein (p) with a median = 0.1 ng/mg p. C-erb B2 values range from 0.02 to 4.53 U/microgram p with a median = 0.43 U/microgram p. Patients with p53 values > 0.1 ng/mg p. had shorter 1 year (20%) and 2 year (13%) -survival times compared to 60% and 20% in patients with p53 values < 0.1 ng/mgp. Patients with cerbB2 > 0.43 U/microgram p. had significantly shorter 1 year (20%) and 2 year survival times (10%) than patients with values < 0.43 U/microgram p (1 year survival = 56% and 2 year survival = 22%; p < 0.05). CONCLUSION: p53 and c erb B2 seem to be valuable prognostic factors in HCC. PMID- 10470183 TI - p53 determination in gynecological carcinomas: immunohistochemistry compared to luminometric immunoassay (LIA). AB - In gynecological carcinomas p53 overexpression has been associated with an aggressive tumor type and shorter overall survival. We compared p53 values analysed by a new quantitative luminometric immunoassay (LIA) applicable on tumor cytosol immunohistochemistry (IH). In 83 breast and 43 ovarian cancers p53 was analysed by LIA and IH (Byk-Sangtec, FRG) and by IH using monoclonal antibodies (MOAB/BP53-Bio Genex, FRG). In breast cancer 32 cases (39%) were immunohistochemically (IH) positive. LIA values ranged from 0.01 to 101 ng/mg protein (p), median (IH negative cases) = 0.208 and median (IH positives) = 0.857 ng/mg p. Using a cut off < 0.2 ng/mg p for a very low expression only 11% (3/27) were IH positive and > 0.8 for very high expression only 16% (3/20) were IH negative. In ovarian cancer values ranged from from 0.01 to 66.1 mg/mg p. 48.8% (21/43) were IH positive, median (IH negatives) = 0.234 and median (IH positives) = 1.43 ng/mg p. At a cut off < 0.2 ng/mg p 25% (3/12) were IH positive and at a cut off > 0.8 ng/mg p only 14% (2/14) were IH negative. CONCLUSION: The quantitative LIA determination of p53 in cytosols of gynecological carcinomas shows a good correlation to immunohistochemistry in cases of very high and very low expression. PMID- 10470184 TI - Prognostic relevance of soluble interleukin-2 receptors in patients with ovarian tumors. AB - Soluble interleukin-2 receptors (sIL-2R) are measurable in the sera of patients with ovarian cancer and several other benign and malignant diseases. However, the function of these sIL-2R is still unclear. Since high levels of sIL-2R are thought to be an indicator of an activated immune system we investigated the correlation of sIL-2R concentration and prognosis of ovarian cancer patients. sIL 2R measurement was performed on the preoperative sera of 130 patients with benign, and 119 patients with malignant ovarian tumors. The IMMULITE sIL-2R assay by DPC Biermann, Bad Nauheim, Germany was used. In ovarian cancer patients sIL-2R concentrations were significantly higher than in those with benign tumors. By defining the 95th percentile of the sIL-2R concentration distribution in patients with benign diseases as the cut-off (1200 U/ml) 35% of the ovarian cancer patients had elevated concentrations. Concentrations of sIL-2R increased with FIGO stage. FIGO-III patients with highly elevated sIL-2R concentrations tended to have better prognosis than those with sIL-2R levels within normal range in contrast to FIGO IV patients. Since sIL-2R concentrations indicate an immunological activation in ovarian cancer patients our data give hints of the possible role of sIL-2R in the assessment of the risk of recurrence in ovarian cancer patients. PMID- 10470185 TI - CASA and Ca 125 in diagnosis and follow-up of advanced ovarian cancer. AB - BACKGROUND: CA 125 is the most important tumor marker in ovarian cancer. Due to its low specificity and the fact that some ovarian malignancies do not produce considerable amounts of CA 125 a combination with the Cancer Associated Serum Antigen (CASA) may reflect more accurately the clinical situation. MATERIALS AND METHODS: CA 125 and CASA determination was performed in sera of 78 patients with advanced ovarian cancer pre- and postoperatively, monthly during chemotherapy and during follow-up care. The cut-off values for CASA were 4 U/ml, for CA 125 35 U/ml and 65 U/ml, respectively. RESULTS: In the detection of advanced ovarian cancer a combination of both tumor markers was superior to the use of either CASA or CA 125 alone. In the follow-up situation CA 125 with the 35 U/ml cut-off showed the highest sensitivity. Both markers had similar prognostic relevance when marker levels three months after surgery were used. CONCLUSION: CA 125 and CASA have similar characteristics in preoperative diagnosis and postoperative follow-up. In clinical situations with inconclusive or negative CA 125 serum values CASA is helpful to improve management of patients with advanced ovarian cancer. PMID- 10470186 TI - Diagnostic value of serum VEGF in women with ovarian tumors. AB - BACKGROUND: Angiogenesis is necessary for growth and invasiveness of malignant tumors. Vascular endothelial growth factor (VEGF) is considered to play a key role in tumor angiogenesis. Few data are available with regard to serum levels of VEGF in patients with ovarian tumors. We investigated the diagnostic value of serum VEGF in patients with ovarian neoplasms. MATERIALS AND METHODS: 61 patients with ovarian neoplasms (41 ovarian carcinomas, 20 cystadenomas) and 20 healthy women were included into the study. VEGF serum concentrations were determined by a commercially available ELISA. RESULTS: Statistical analysis revealed significant differences in VEGF serum values of ovarian cancer patients vs. healthy controls or patients with cystadenomas. No difference could be seen between serum levels of healthy controls and women with benign ovarian tumors. For ovarian cancer patients vs. normal controls a sensitivity of 71% and a specificity of 65% resulted. The sensitivity and specificity of cancer patients vs. patients with benign neoplasms were 71% and 65%, respectively. CONCLUSION: Our results suggest that VEGF has potential as a serum marker with diagnostic relevance in ovarian neoplasms. PMID- 10470188 TI - Prognostic relevance of the endothelial marker CD 34 in ovarian cancer. AB - BACKGROUND: Tumour angiogenesis and microvessel density are of prognostic significance in several human neoplasia. To investigate how tumour vascularity correlates with disease-free survival microvessel density was assessed in 38 patients with ovarian cancer using the highly specific endothelial marker CD 34. METHODS: Representative specimens were obtained and stained using monoclonal CD 34 antibodies. The microvessels were quantified at 200x and 400x magnification in the most active areas of neovascularisation. Degree of angiogenesis was correlated with histologic tumour type, grading, and tumour stage. Furthermore, survival was calculated using Kaplan-Meier analysis and results again correlated with the microvessel count. RESULTS: No correlation was found between microvessel density, histologic type, grading, and FIGO stage. Patients with a vessel count more than 40 (200x magnification) had a statistically significant lower overall survival (p = 0.022). CONCLUSION: Our results indicate that CD 34 is a useful marker in determining tumour neovascularisation which might be of prognostic relevance in patients with ovarian cancer. PMID- 10470187 TI - Prognostic significance of CA 125 and TPS levels after chemotherapy in ovarian cancer patients. AB - The analysis of survival data of patients with epithelial ovarian cancer proved that both CA 125 and TPS were good markers for clinical outcome prediction. Patients receiving chemotherapy were analyzed for 2-year overall survival (OS). Kaplan-Meier survival analysis showed highly significant differences in OS between patients with stage I+II (survival for 2 years 68%) and stage III+IV (survival for 2 years 33%; p = 0.0008). CA 125 levels above or below 35 kU/I and TPS levels above or below 80 U/l after 3 chemotherapy courses were not significantly correlated with OS in stage I+II patients (p = 0.06 respectively 0.07). However, in the subgroup of patients with stage III+IV the cut-off levels of CA 125 and TPS were excellent discriminators of OS: With CA 125 levels below the cut-off 52% of the patients survived, while with CA 125 levels above the cut off only 13% survived (p < 0.0001). With TPS levels below the cut-off 49% of the patients survived, while with levels above the cut-off only 19% of the patients survived (p < 0.0001). In the subset of patients with CA 125 levels less than 35 kU/I after 3 chemotherapy courses (n = 50) analysis of their TPS levels allowed further discrimination of the prognostic significance. With TPS levels below the cut-off 63% of the patients survived, while 35% of the patients survived with TPS levels above the cut-off. The sum value of CA 125 and TPS cut-off values (115) as discriminator correlated even better with survival rate: With levels below this sum value 63% of the patients survived, while this was only 17% with sum values above the summed cut-off level (p = 0.0004). The extent to which the tumor was removed at operation also correlated with the 2 years survival rate. None of the patients with a staging laparotomy (n = 10) showed a 2-years survival. The difference in OS between patients with complete debulking and partial debulking was significant: OS 51% versus 23% (p = 0.027). Prognosis was not significantly correlated with histological type. PMID- 10470189 TI - Fluctuations of tumor markers in heart failure patients pre and post heart transplantation. AB - BACKGROUND: Elevated plasma levels of tumor markers may be caused by diseases other than malignancy, i.e. kidney, liver or circulatory disturbances. These conditions are not well defined, especially since there are only sparse reports on fluctuations of tumor markers related to cardiac function. PATIENTS AND METHODS: During our routine pre- and postoperative follow-up tumor marker determinations in heart failure patients were made in order to screen for possible occult neoplasm's which may either be a contraindication or a sequela of heart transplantation. The markers CA 12-5, CEA, CA 19-9, CA 72-4, TPA, TPS and CYFRA 21-1 were determined at three month intervals, besides clinical examination and hemodynamic measurements in a total of n = 118 patients pre- and n = 74 patients post heart transplantation. RESULTS: The results were grouped according the clinical status (NYHA-stage 1-4): CA12-5 (29.4 +/- 40.63 omega 151, 174 +/- 345 and 491 +/- 633 U/ml, p < 0.001 between all groups) and TPS (64 +/- 32, 118 +/- 153, 163 +/- 311 and 181 +/- 232 U/ml, p = 0.06 between all groups) were increasingly elevated in NYHA stages 1, 2, 3 or 4 respectively. A direct correlation to right atrial pressure (r = 0.41, p < 0.0001) and pulmonary capillary wedge pressure (r = 0.27, p < 0.001) was only found for CA 12-5. After heart transplantation a normalization of elevated pre-OP levels could be found. Comparable to heart failure patients poor graft function was also associated with elevated levels of CA 12-5 (113 +/- 99 vs 21.6 +/- 31 U/ml, p < 0.0001), CA 72-4 (8.4 +/- 3 vs 3.6 +/- 4, U/ml p = 0.03) and TPS (154 +/- 133 vs 66 +/- 28 U/ml, p < 0.001). The individual time course of the markers, especially of CA 12-5, correlated nicely to clinical events and hemodynamic measurements in some patients. Another finding was that CYFRA 21-1 levels were correlated to renal function. CEA, CA 19-9 and CYFRA 21-1 serum levels were not influenced by circulatory disturbances. CONCLUSION: We concluded that the tumor markers CA 12-5 and TPS (but not CEA, CA 19-9 and CYFRA 21-1) are associated with congestion and the clinical course of heart failure and HTx patients. These "nonspecific" changes have to be considered when tumor markers are determined in cancer patients with heart failure. Whether CA 12-5 blood levels may yield additional prognostic information in the management of cardiovascular patients has to be determined in further studies. PMID- 10470190 TI - IMMULITE OM-MA assay: a useful diagnostic tool in patients with benign and malignant ovarian tumors. AB - Measurement of CA125 is well established in the diagnosis and follow-up of ovarian cancer. CA125 determination is usually performed with the monoclonal antibodies OC125 and M11. For the IMMULITE OM-MA chemoluminsence assay (DPC Biermann, Bad Nauheim, Germany) the monoclonal antibody OV185 is used. We analysed CA125 serum concentrations of patients with benign and malignant ovarian tumors with the IMMULITE OM-MA assay by defining a cut-off at 90% specificity level. Sera of 130 patients with benign and 119 patients with malignant ovarian tumors at different FIGO stages were analysed. By calculating a cut-off of 46 U/ml, in 87 ovarian cancer patients, elevated CA125 concentrations were detected (78% sensitivity). There were no significant differences between CA125 concentrations of tumors of different histological type. The measurement of CA125 by IMMULITE OM-MA is a simple and sensitive procedure. PMID- 10470191 TI - The course of the urinary DPD-crosslink secretion in metastatic breast cancer--a possibility for response assessment. AB - BACKGROUND: Urinary deoxypyridinoline (DPD)-crosslinks have been shown to be a highly specific parameter for type I collagen metabolism. MATERIALS AND METHODS: In a prospective breast cancer study, urine samples were collected in after-care patients and in patients with bone metastases. DPD-crosslinks were measured every three weeks using a fully automated chemiluminescence immunoassay. Bone metastases were confirmed by bone scan and/or x-ray, and were followed-up over six months. To validate the test, a receiver operating characteristics (ROC) curve was set up to find the DPD cut-off concentration which separates patients with no evidence of disease (NED) from patients with bone metastases. RESULTS: 73 breast cancer patients (41 with NED, 32 with bone metastases) were included into the ROC analysis. At a DPD cut-off value of 8 nmol/mmol creatinine, we found the best sensitivity (84.4%) for the detection of bone metastases with a specificity of 70.7%. Patients with stable bone disease under intravenous pamidronate treatment (90 mg q3w) and specific therapy had a significant (p = 0.007) fall of the DPD-crosslinks in comparison to the progressive subset with 72.7% falling below 8 nmol/mmol. CONCLUSIONS: We conclude that the net bone turnover is not increased at a DPD-crosslinks elimination < 8 nmol/mmol. PMID- 10470192 TI - Tumour markers CEA and CA 15-3 as Prognostic factors in breast cancer--univariate and multivariate analysis. AB - Tumour markers are putative prognostic indicators for patients with breast cancer, but have not been elevated independently by multivariate analysis in a large patient number. In 550 patients with breast cancer without known metastases the levels of the serum tumour markers CEA und CA 15-3 were determined preoperatively and during follow-up. The prognostic relevance of these markers for recurrence (n = 128/487) and death of disease (n = 55/550) was evaluated in relation to established prognostic factors. In univariate analysis tumour size, lymph nodes, histological grading, age, hormone receptors, preoperative value of CEA (cut-off 2 ng/mL) and CA 15-3 (cut-off 25 U/mL) and their decrease of more than 33% within seven months after operation were significant for relapse. The results for death of disease were similar except for age. In multivariate analysis tumour size, lymph nodes and decrease of CEA > 33% (p < 0.001) were independent prognostic factors for recurrence. For overall survival tumour size, lymph nodes, histological grading and preoperative levels of CEA > or = 2 ng/mL (p = 0.038) and of CA 15-3 > or = 25 U/mL (p = 0.007) were independent prognostic factors. Pre- and postoperative values of the tumour markers CEA und CA 15-3 are strong independent prognostic factors for relapse and survival in breast cancer patients. PMID- 10470193 TI - C-erbB-2, CEA and CA 15.3 serum levels in the early diagnosis of recurrence of breast cancer patients. AB - C-erbB-2, CEA and CA 15.3 serial serum determinations were performed in 250 patients (follow-up: 1-4 years, mean 2.5 years) with primary breast cancer and no evidence of residual disease (NED) after radical treatment (radical mastectomy or simple mastectomy and radiotherapy). Ninety-five patients developed metastases during follow-up. RESULTS: Abnormal c-erbB-2, CEA and CA 15.3 serum levels (> 20 U/ml, > 10 ng/ml or > 60 U/ml, respectively) prior to diagnosis were found in 28.4%, 31.6% and 46.3% of the 95 patients with recurrence, with a lead time of 4.2 +/- 2.4, 5.0 +/- 2.5 and 4.6 +/- 2.7 months, respectively. One of the tumor markers was the first sign of recurrence in 69.5% of the patients. Tumor marker specificity was 100% with levels lower than the cut-point in all 155 patients without recurrence. Tumor marker sensitivity was clearly related to the site of recurrence, with the lowest sensitivity found in locoregional relapse and the highest in patients with liver or bone metastases. C-erbB-2 sensitivity in early diagnosis was significantly higher in patients with c-erbB-2 overexpression in tissue (10/12, 83.3%) than in those without overexpression (1/34, 2.9%) (p = 0.0001). Likewise, higher levels of both, c-erbB-2 and CA 15.3 at diagnosis of recurrence, higher sensitivity in early diagnosis of relapse and a higher lead time were found in PgR+ patients (CA 15.3) or in PgR- patients (C-erbB-2) (p < 0.015). In conclusion, tumor markers are useful tools for the early diagnosis of metastases, being the first sign of recurrence in 69.5% of patients with relapse (76.3% in patients with metastases). PMID- 10470195 TI - Immunohistochemical analysis of prostate-specific antigen does not correlate to other prognostic factors in breast cancer. AB - BACKGROUND: Prostate-specific antigen (PSA) is found in 30-40% of female breast tumors, as recently described. Diamandis and co-workers could demonstrate that PSA correlate significantly to a relapse-free survival and lower tumor stages in patients with breast cancer using a time-resolved immunofluorometric analysis. The presence of PSA in these tumors seems to reflect a favourable prognostic marker for that disease. The aim of our study was to evaluate the role of PSA as a prognostic factor in breast cancer using an immunohistochemical technique. MATERIALS AND METHODS: The PSA immunoreactivity of tissues from one-hundred women with malignant breast tumors was correlated to tumor staging, histomorphological tumor type, and biochemical estrogen and progesterone receptor content. Additionally, survival analysis was performed according to Kaplan and Meier. RESULTS: 49% of the tumors revealed positive staining for PSA. No significant correlation between PSA and the other parameters, or the mean survival time (PSA pos.: 5.3 years, PSA neg.: 5.4 years) could be demonstrated. CONCLUSIONS: As there were no significant correlations between PSA and other prognostic markers, PSA detected by immunohistochemistry seems not to be helpful in prognostic evaluation of breast cancer. PMID- 10470194 TI - Carcinoembryonic antigen in tissue and serum from breast cancer patients relationship with steroid receptors and clinical applications in the prognosis and early diagnosis of relapse. AB - From July 1982 to August 1994, CEA levels were determined in 298 mammary tissue samples (30 benign, 242 primary breast cancer and 26 metastatic breast cancer). CEA serum levels were also evaluated in 30 patients with benign diseases, 153 patients with primary breast cancer and 26 patients with metastases. CEA tissue levels in both pellet and cytosol were significantly higher in samples from cancerous than from non malignant tissues (p < 0.0001), and higher in the pellet than in the cytosol (p < 0.0001). CEA in the pellet and cytosol were related to steroid receptors, with the highest levels being observed in ER+/PR+ tumors (p < 0.001). They were, however, not related to other pathological parameters such as tumor size or nodes. There was a correlation between CEA pellet and CEA serum in both patients with primary or metastatic tumors, with significantly higher CEA serum levels in patients with CEA pellet positivity than in those with CEA pellet negativity. CEA serum levels were a prognostic factor (DFS and OS) in the whole group as well as in node-positive and node-negative breast cancer patients. This prognostic value was only found in patients with CEA pellet positivity. In the follow-up of 143 NED patients, abnormal CEA serum levels rose prior to the diagnosis of relapse in 73% (29/40) of CEA pellet+ patients with distant recurrences but in only 9% (2/22) of CEA pellet- cases (p < 0.0001). In summary, CEA evaluation in tissue improves the CEA clinical application, selecting those patients whose serum CEA will be useful in the prognosis and early diagnosis of recurrence. PMID- 10470196 TI - Predictive value of CEA and CA 15-3 in the follow up of invasive breast cancer. AB - BACKGROUND: The clinical usefulness of tumor markers in the follow-up care of invasive breast cancer is controversial. METHODS: In 1228 serum samples of 664 women with history of breast cancer, the diagnostic accuracy and predictive power of CEA and CA 15-3 for the detection of disease relapse was determined prospectively by analyzing the clinical course for at least 6 months after the measurement of the tumor markers in 1994. RESULTS: A total of 76 patients relapsed during the period of study. The diagnostic accuracy was 83% for CEA and 88% for CA 15-3. CEA and CA 15-3 had a positive predictive value of 27% and 47% as well as a negative prediction of 91% and 93%, respectively. CONCLUSIONS: The low positive predictive value and sensitivity of these tumor markers clearly limit their clinical utility. Therefore, the effectiveness of routine determinations during the follow-up seems questionable. PMID- 10470197 TI - Comparison of the tumor associated proteases cathepsin D (CATH D) and urokinase type plasminogen activator (uPA) in cytosols of human breast cancer patients. AB - As a matter of routine, prepared cytosols of human primary breast cancer specimens (n = 230) are analysed for both CATH D and hormone receptor status (ER,PR). Retrospectively, uPA was determined in the samples. The selection criterion for the retrospective analysis was the possibility of a longitudinal follow-up of the patients. All tumor stages were included, but the main emphasis was on lower tumor stages (T1, and T2) as well as nodal negative stages (N0). The results of the hormone receptor status (ER,PR) were: ER+PR+ 66.37%; ER+PR- 10.18%, ER-PR+ 10.18%, ER-PR- 13.27%. The CATH D results ranged from 5 to 246 pmol/mg protein. 30.97% of these results rose above 50 pmol/mg protein (positive), 42.48% were below 35 pmol/mg protein (negative). The uPA results ranged from 0.05 to 3.74 ng/mg protein. 27.43% of the uPA results rose above 0.71 ng/mg protein (positive), 58.85% were below 0.53 ng/mg protein (negative). Raised results of uPA are distinct in the higher tumor stages (T1N0 > T2N0 > T2N1 > T4Nx). Although the CATH D and uPA measurements showed similar results (positive/negative distribution) in the general survey, the confirmity of both factors is rather limited if it is focused to single cases. Between CATH D and uPA there was a confirmity of 50% (T1N0) of 45.5% (T2N0) respectively in the range of positive results, and there was a confirmity of 60.3% (T1N0) and 65.8% (T2N0), respectively, in the range of negative results. Differences were seen in 19.2% (T1N0) and 22.7% (T2N0), respectively, in the range of positive results, and in 20.6% (T1N0) in 1508% (T2N0), respectively, in the range of negative results. The prospectively diagnostic value of these is still under observation. PMID- 10470198 TI - Control of hepatic parameters in renal cell carcinoma (RCC) by interleukin-6 (IL 6)? AB - RCC can go hand-in-hand with an elevation of various hepatic proteins. An interrelationship between the IL-6 titer, C-reactive protein (CRP) and the blood sedimentation rate (BSR) has already been proven. The aim of the present study was to study 1) the possibility of differentiating between healthy and RCC patients via IL-6 in the serum and 2) the relationship of IL-6 to hepatic parameters {alkaline phosphatase (AP), gamma-glutamyltransferase (gGT), serum proteins (E'p)} and the usual clinical prognostic parameters (tumor grading, staging). Serum analysis of 38 healthy patients via ELISA (DPC-Biermann, Germany) showed normal values of 1.2 ng/ml for IL-6, with a standards deviation of +/- 1.7 and a peak concentration of 3 ng/ml (specificity: 95%). In 20 RCCs there were IL 6 titers of 10.7 ng/ml +/- 6.56 in the pre-operative serum. The sensitivity of IL 6 was about 90%. The difference was statistically significant (p < 0.0001, Wilcoxon test). For IL-6 there was a positive correlation with the BSR (1-hour value: r = 0.7; 2-hour value: r = 0.6), CRP (r = 0.85), E'p (r = 0.6), and gGT (r = 0.6). No correlation was found between AP, the Robson stage, grading, and IL-6. IL-6 is potentially suitable for differentiating between healthy and RCC patients but is not tumor specific. IL-6 has a strong correlation with all laboratory values which were analyzed except AP thus there is considerable evidence for a cytokine (IL-6) control of the hepatic changes. Since some of the above-named laboratory parameters have prognostic relevance, IL-6 can be regarded as a cumulative prognostic parameter. PMID- 10470199 TI - Marker for renal cell carcinoma (RCC): the dimeric form of pyruvate kinase type M2 (Tu M2-PK). AB - OBJECTIVE: The evaluate a potential tumor marker for RCC. Tumor formation is generally linked to an expansion of glycolytic phosphometabolite pools and aerobic glycolyticflux rates. To achieve this, tumor cells generally overexpress a special glycolytic isoenzyme, termed pyruvate kinase type M2. To establish the expansion of phosphometabolite pools pyruvate kinase switches between a tetrameric form with high phosphoenol-pyruvate (PEP) affinity and a dimeric form with a lower PEP affinity. The dimeric form is predominant in all tumors that have been investigated and has been termed TuM2Pk. MATERIALS AND METHODS: We studied: a) the expression of TuM2Pk in RCC by immunohistochemistry using a monoclonal antibody recognizing only the mono- or dimeric form of pyruvate kinase, b) the stability of TuM2Pk in serum by measuring TuM2Pk in 3 patients at different times after taking blood with a two-site immunometric assay, c) the a circadiane rhythm of TuM2Pk in blood by measuring levels every 4 hours in 5 patients, d) TuM2Pk- expression in serum (see 2.) in 5 patients by taking blood from tumor-side vena renalis compared to peripherally blood, e) TuM2Pk (see point 2.) in 40 RCC-patients comparing the results with 39 healthy persons and clinical data of RCC, f) the influence of wound healing to TuM2Pk by measuring serum levels during a period of more than 12 weeks in 6 patients, g) the individual follow up of 4 patients with RCC stage Robson III for more than 2 years. Comparing TuM2Pk-levels to findings of staging by computed tomography. RESULTS: The isoenzyme TuM2Pk could be demonstrated in RCC and their metastases by immunohistochemistry with a monoclonal antibody specific for pyruvate kinase type M2. In normal kidney cells pyruvate kinase type M2 is not detectable. The stability of TuM2Pk was studied in the serum within 30 minutes. No circadian rhythm was found. Most serum TuM2Pk comes from tumor. Serum evaluation in 39 healthy persons was used to determine normal values, with an upper concentration of 28 U/ml of TuM2Pk (95% percentile of normal healthy persons). Serum evaluation in 40 RCC showed a significant difference to healthy persons and a positive correlation with Robson stage and grading No correlation of TuM2Pk was found with histopathological cell type of tumor diameter. After radical nephrectomy normalization of TuM2Pk level was found within 11 weeks in all localized RCC. Continuously elevated serum levels were seen in metastatic RCC. Individual follow up seems to be possible. CONCLUSION: Initial discrimination is not possible between localized and metastasized RCC using TuM2PK; however, it is possible to differentiate between benign and malignant renal processes; the specificity under these circumstances is 75%. After successful surgery of localized RCC, an elevated TuM2Pk will be normalized within 11 weeks, and will be remain elevated or will increase again in case of RCC-relapse or metastasis. Thus TuM2Pk would appear to be a useful marker for RCC detection and follow-up. PMID- 10470200 TI - Immunohistological investigations of carcinoembryonic antigen (CEA) in urothelial carcinomas. AB - CEA is discussed as a potential tumour marker for carcinomas of the urothelium. However, investigations by means of polyclonal antibodies have shown various cross reactions which are not to be expected with monoclonal antibodies (MAb). Immunohistochemistry was done with the MAb anti-CEA BW 431/26 (Behring), N 1522 (Dako), and C 7292 (Sigma) on 37 carcinomas of the urothelium (grades 1-3). The MAb BW 431/26 showed the best results concerning specificity and sensitivity. In general, immunohistological investigations demonstrated negative or moderate staining reactions. Positive reactions were seen in 32% (12/37) of the carcinomas. Staining intensity for CEA correlated with differentiation grade. On the whole, a maximum of 5% of the tumour cells were CEA positive. Our results indicate that monoclonal anti-CEA antibodies are not usefull as a tumour marker for urothelial carcinomas. For the suitability of CEA for in vitro and for in vivo diagnosis a threshold of CEA positive tumour cells has to be defined. PMID- 10470201 TI - The pyruvate kinase isoenzyme tumor M2 (Tu M2-PK) as a tumor marker for renal carcinoma. AB - Various isoforms of the glycolytic enzyme pyruvate kinase are expressed in different cell-types. One of these isoforms, the type Tu M2-PK, is strongly overexpressed by tumor cells and released into body fluids. The concentration of Tu M2-PK in body fluids can be quantitatively determined by a commercially available enzyme-linked immunosorbent assay (ELISA)-kit. Using this kit, the Tu M2-PK concentration was measured in EDTA-plasma of 64 patients with renal carcinoma and 10 patients suffering from nephritis. The ranges of the Tu M2-PK concentrations of the two groups did not overlap, indicating a highly significant discrimination of renal carcinoma and benign renal diseases. Furthermore, the Tu M2-PK-concentration in EDTA-plasma correlates strongly with the Robson tumor stage of the 64 patients. The present results indicate that the Tu M2-PK might be the first tumor marker which could be an excellent complementation of the diagnostic program for renal carcinoma. PMID- 10470203 TI - Serum tissue polypeptide antigen (TPA): monoclonal or polyclonal radio immunometric assay for the follow-up of bladder cancer. AB - Tissue polypeptide antigen (TPA) is a serological tumor marker, measuring cytokeratin 8, 18 and 19, used in the follow-up of non squamous epithelium- and derived neoplasms. It has been demonstrated that TPA is reliable in the monitoring of the efficacy of a curative or palliative treatment of bladder cancer. Recently, a monoclonal antibody-based assay for TPA (TPA-M) has been developed, which seems to be equivalent to the polyclonal-based assay. The aim of the study was to assess the superiority of the monoclonal to the polyclonal test in patients with bladder carcinoma. The value of tissue polypeptide antigen was therefore measured both with TPA and TPA-M IRMA. A correlation coefficient of 0.96 was obtained. Precision testing showed a lower overall variability of TPA-M. Since both tests correlate well and TPA-M testing is more precise, faster and easy to perform, we conclude a superiority of TPA-M and advise the monoclonal test as best suited for clinical use in the follow-up of bladder cancer patients with poorly differentiated superficial, locally advanced or systemic disease after curative or palliative therapy. PMID- 10470204 TI - BTA-TRAK--a useful diagnostic tool in urinary bladder cancer? AB - During recent years the BTA-TRAK-assay (Bard Diagnostics, Redmont, USA) has been described in several investigations to be of clinical utility for patients suffering from bladder cancer. In a prospective study we investigated over four months the voided urine samples of all consecutive patients undergoing cystoscopy independent of their clinical background (n = 244) with the BTA-TRAK-assay. With a specificity of 95% for benign urological diseases (cut off: 1300 U/mL) we found a sensitivity of 13% for active bladder tumours. Using healthy individuals as a reference group (cut off: 40 U/mL) we found a sensitivity of 56% (specificity 67%). Using the cut off value recommended by the manufacturer (14 U/mL) a specificity of 54% and a sensitivity of 62% was found. For patients without relapse (NED) versus patients with active bladder tumours we got a specificity of 55% and a sensitivity of 62%. Due to an insufficient specificity and sensitivity the BTA-TRAK-test is not able to replace cystoscopy nor to improve existing diagnostic strategies in bladder cancer. PMID- 10470202 TI - The clinical value of CYFRA21-1 in bladder cancer patients: Egyptian experience. AB - There are a wide variety of tumor markers now available that proved to be of value in the management of cancer patients. Of these markers, tissue polypeptide antigen (TPA) and tissue polypeptide specific antigen (TPS) are well known in the field of bladder cancer. TPA was found to be a mixture of cytokeratins 8, 18 and 19 and recent investigations proved that TPS is keratin 18. The aim of the present study was to assess the clinical value of urinary cytokeratin 19 (CYFRA21 1) in the differential diagnosis between bladder cancer and benign urinary tract diseases represented by bilharziasis. Two hundreds and seventy individuals were included in the present study: 186 with bladder cancer representing the different stages and grades, 44 with urinary tract bilharziasis and 40 normal healthy controls. CYFRA21-1 was evaluated in 24-hour urine samples by ELISA using the automatic set supplied by Boehringer Manheim, Manheim, Germany (ES 300). Results of this study revealed significant elevation of CYFRA21-1 in bladder cancer followed by bilharziasis. 82.3% (153/186) of bladder cancer patients and 11.4% (5/44) of bilharzial patients exhibited CYFRA21-1 levels above the upper limit of the control group (3 micrograms/24-hr). CYFRA21-1 was more sensitive in advanced than early stages of bladder cancer and in patients with positive than those with negative lymph nodes, but association of tumor with bilharziasis did not markedly affect its level. PMID- 10470205 TI - NMP22: a sensitive, cost-effective test in patients at risk for bladder cancer. AB - BACKGROUND: This study was designed to determine the clinical utility of NMP22 as a urinary marker for the early detection of transitional cell carcinoma (TCC) of the bladder in patients with hematuria or other indications for risk of malignancy. Its utility will be measured by sensitivity and specificity estimates as compared to cystoscopy. Since urine cytology is normally collected in this population of patients, it will also be analyzed and compared to cystoscopy. MATERIALS AND METHODS: Each patient submitted a single voided urine which was stabilized with the NMP22 urine collection kit or preserved in the appropriate cytology medium for cytopathologic testing. All patients provided the urine samples before cystoscopic exam. Of the 146 patients, there were 43 patients with microscopic hematuria and 13 with gross hematuria. Other indications for cystoscopy included unexplained or medically refractory voiding. There were 8 patients with biopsy confirmed bladder cancer and 138 patients with benign conditions of the bladder. RESULTS: The median NMP22 value for the bladder cancer malignancies was 27.8 U/mL (95% Confidence interval: 10.5-32.1 U/mL). The median NMP22 value for the benign conditions of the bladder was 3.25 U/mL (95% Confidence interval: 2.5-3.8 U/mL). The urinary NMP22 values from the bladder cancer group was statistically different (p < .000001 Mann-Whitney U test) than the NMP22 values in the benign conditions group. Using a reference value of 10.0 U/mL, the sensitivity of NMP22 was 100% with a specificity of 90%, while cytology had a sensitivity of 25% and a specificity of 100%. Due to its high negative predictive value, using NMP22 alone could have eliminated 124 cystoscopies with total savings ranging from $24,824 to $63,264 depending on the type of insurance carrier. CONCLUSIONS: This study indicates that urinary NMP22 is a useful, cost effective marker for the early detection of bladder cancer. PMID- 10470206 TI - The biologic lower detection limit of six ultrasensitive PSA assays. AB - BACKGROUND: The benefit of monitoring patients with prostate cancer (PCA) by ultrasensitive measurement of prostate specific antigen (PSA) is frequently discussed. Usually, the analytic lower detection limit of an ultrasensitive assay is determined by the manufacturer. As the analytic lower detection limit does not take into account interfering factors of human serum, the biologic lower detection limit, which is defined as PSA concentration detected in PSA-free human serum, plus 3 standard deviations, is of greater interest. MATERIALS AND METHODS: We investigated the biologic lower detection limit of six ultrasensitive PSA assays. Sera from 15 men with bladder cancer after radical cystoprostatectomy and from 30 healthy women were applied. Hence, we expected no PSA of prostatic origin. RESULTS: The biologic lower detection limit obtained using these sera was up to 30 fold higher (men, 0.29-0.63 ng/ml; women, 0.03-0.69 ng/ml) than the analytic lower detection limit (0.01-0.09 ng/ml). CONCLUSIONS: PSA measurement in sera obtained from men without prostate and women results in PSA values above the ultrasensitive range. Therefore, advantages provided by ultrasensitive PSA measurement in monitoring PCA patients after radical prostatectomy are limited. PMID- 10470207 TI - Age related changes of free and total prostate specific antigen in serum. AB - PSA in serum exists in several molecular forms. The amount of free and total PSA and ratio are discussed to be useful to increase the ability of PSA to distinguish prostate cancer from benign prostatic hyperplasia. Therefore, we tried to characterize the age related changes of free and total PSA in a German rural population. METHODS: Serum samples were obtained from 697 men. All study participants were between 20 and 79 years old and had no clinical evidence of prostate cancer by serum PSA, digital rectal examination and transrectal ultrasonography. The sera were kept frozen at -20 degrees C until analysis and were assayed within 3 months after sampling. Free and total PSA values were determined employing a chemiluminescent enzyme immunoassay (Immulite PSA and Freies PSA, DPC Biermann). RESULTS: Free PSA, total PSA and the free-to-total PSA ratio (f-/t-ratio) demonstrated a correlation with patient age (r = 0.855, p = 0.0141; r = 0.857, p = 0.0137; r = -0.788, p = 0.0352). Employing the median and the 95% percentile the age-specific reference ranges for free and total PSA were calculated. For the f-/t-ratio the median and the interquartile range (i.e. 25th 75th percentile) were calculated. CONCLUSION: In confirmation of a recent report, we found a direct correlation of free PSA, total PSA and the f-/t-ratio with age, whereas free and total PSA increases, the f-/t-ratio decreased with advancing age. PMID- 10470208 TI - Critical aspects related to the interpretation of the free-to-total PSA-ratio. AB - INTRODUCTION: The number of assays available for the measurement of total and free PSA is increasing. As different methods can determine different PSA concentrations as well as different free-to-total PSA ratios in identical serum samples, the cut-offvalue for the ratio still needs to be determined. METHODS: 114 sera from patients with histologically confirmed benign prostatic hyperplasia (BPH; n = 58) and cancer of the prostate (CaP; n = 56) were analyzed with two different assays. Free PSA (free), total PSA (total) and the free-to-total- PSA ratio (ratio) were determined employing Enzym-Test PSA und freies PSA (Boehringer Mannheim, Germany) and Immulite PSA und freies PSA (DPC Biermann, Bad Nauheim, Germany) RESULTS: The statistical results are tabulated below: [table: see text] CONCLUSION: Direct comparison of the two assays revealed a high statistical correlation (r = 0.94-0.99) for free and total PSA. In contrast, the ratio of the two assays was not as reproducible (r = 0.81-0.83). This result indicates that the reference range for the ratio is dependent on the assay employed and an that uncritical use of an applied reference range can be counter-productive. PMID- 10470209 TI - Estimation of total PSA with a supersensitive PSA-assay during neo-adjuvant chemotherapy of prostate cancer before radical resection of prostate. AB - 43 patients with untreated and clinically localised prostate carcinoma (cT1 3NoM0) were submitted to neo-adjuvant complete androgen deprivation treatment and radical prostatectomy. Hormonal treatment was given until PSA (supersensitive Immulite 3rd generation assay) reached a value of < 0.1 ng/ml or the nadir value. The resultant mean duration of treatment was 6 months (3-22 months). 93% of our patients reached a PSA value of 0.1 ng/ml or below. The nadir of 6 patients was between 0.1-0.3 ng/ml. In one case it remained above 0.3 ng/ml. 82% had a measurable hypoechoic lesion on initial transrectal ultrasound. 84% of these became smaller, 7.5% remained unchanged and 8.5% increased. Of the patients with cT1-2 tumours 2% had a positive margin and in 28% of the specimens no tumour tissue could be detected at the time of prostatectomy. Of the initial patients with epithelial cells in bone marrow only 14% remained positive after controlled induction and all of them had fewer cells than before. Five patients relapsed showing raising PSA values > 0.1 ng/ml in the follow-up. In this group the mean PSA and the time of nadir was significantly higher (0.15 ng/ml) than the mean PSA at the time of nadir of the rest of the group (0.06 ng/ml). Supersensitive PSA assays like Immulite 3rd generation offer a valuable tool to collect data for clinical evaluation to optimise therapy of prostate carcinomas. PMID- 10470210 TI - Supersensitive PSA-analysis after radical prostatectomy: a powerful tool to reduce the time gap between surgery and evidence of biochemical failure. AB - BACKGROUND: Using three commercially available, unmodified PSA assays, 917 sera of 355 patients after radical prostatectomy underwent native and lyophilisation concentrated PSA-detection to evaluate the benefit of serum concentration as a tool to increase sensitivity for earlier detection of recurrent prostate cancer after RP. MATERIALS AND METHODS: We evaluated Abbott IMX, Tosoh ALA-600 and DPC Immulite Assay in the follow-up of 355 patients. Mean follow-up time is 374 days (43-2057). All sera underwent native analysis and 4 fold- lyoconcentration and subsequent triplicate standard analysis on each assay. Evaluation of native and concentrated sera: A PSA-value of > or = 0.10 ng/ml was positive. Sera reading < 0.10 ng/ml were considered negative. In 4-fold concentrated sera this means a calculated increase of sensitivity to > or = 0.025 ng/ml. The average day of detection of a first positive signal either in native or concentrated sera for each assay was calculated as well as the time showing the average day difference of earlier detection in the lyoconcentrated sera for each assay. 20 Female sera were run and consistently read zero values in native and concentrated sera. RESULTS: In 355 patients, the number of PSA-positive patients were 58, 65 and 62 on Abbott, Tosoh and Immulite, respectively. 17/58 (29.3%), 19/65 (29.2%) and 20/62 (31.7%) patients could be identified earlier in lyoconcentrated than in native sera. A mean time advantage of 308-336 days was found. No patient who was positive according to supersensitive criteria had a negative supersensitive result, in his later follow-up. 17/19 (89.4%) patients in the Tosoh, 17/20 (85%) in the Immulite and 10/17 (58.8%) in the Abbott were identified as PSA-positive within one year after RP. CONCLUSION: PSA was positive in lyoconcentrated sera a mean of one year earlier than in native sera The maximum time advantage of lyoconcentration was 862 days, indicating PSA-recurrence 2.3 years earlier than standard analysis. 58.8-89.4% of patients with biochemical evidence of PSA recurrence in supersensitive analysis were found within one year after RP. Lyoconcentration increases PSA-signals reliably in different assays. It can be performed as a quick routine procedure. PMID- 10470211 TI - Prostate-specific antigen (PSA) and radiotherapy in locally confined prostate cancer. AB - BACKGROUND: During the last years, prostate-specific antigen (PSA) has been established as the most important tumor marker for prostate cancer. The aim of our study was to evaluate the response of PSA during and after radiotherapy of prostate cancer. PATIENTS AND METHODS: From 1/95 to 6/97, 26 patients were irradiated for locally confined prostate cancer (11/26 patients primarily and 15/26 patients after radical prostatectomy). Radiation therapy was carried out with a linear accelerator with a total dose of 60 Gy to the prostate bed. PSA values were measured immediately before, weekly during radiation therapy and every three months after in the usual follow-up pattern. RESULTS: During radiotherapy, median PSA values fell and continued to fall after radiotherapy. The median PSA half-life was reached 3 months after irradiation, the PSA nadir after 15 months. 5 patients with increasing PSA levels relapsed. DISCUSSION: During and after radiotherapy, PSA kinetics are low with a median half-life of 3 months in our patient population. Rising PSA profile indicates progression of tumor. PMID- 10470212 TI - Cytokeratin markers in patients with prostatic diseases. AB - OBJECTIVES: Recently tissue polypeptide specific antigen (TPS), a cytokeratin 18 marker, was described to be discriminative between cancer of the prostate (CaP) and benign prostatic hyperplasia (BPH). Cyfra 8/18, a marker which recognizes both cytokeratin 8 and 18 fragments, is thought to improve sensitivity and specificity of TPS. In our study we investigated the ability of the TPS and cyfra 8/18 serum concentration to discriminate between patients with clinically localized CaP and BPH. METHODS: Serum levels of TPS and Cyfra 8/18 were determined in patients with untreated CaP (pT1-3pNoMo: n = 11) and BPH (n = 22). The TPS and the Cyfra 8/18 concentrations were correlated to the prostate specific antigen (PSA) serum concentration. RESULTS: Median TPS concentration was 45.3 U/L in CaP-patients and 54.8 U/L in BPH-patients. This difference is statistically not significant (p = 0.2). Median Cyfra 8/18 level was 0.64 ng/mL in CaP-patients and 0.57 ng/mL in BPH-patients. This difference is statistically not significant (p = 0.91). Furthermore no correlation with PSA levels could be established (TPS: r = -0.13; Cyfra 8/18: r = 0.17). CONCLUSION: In contrast to recent reports we found both cytokeratin markers, TPS as well as Cyfra 8/18, to be non-discriminative parameters in CaP and BPH. PMID- 10470213 TI - Metastatic workup of patients with prostate cancer employing alkaline phosphatase and skeletal alkaline phosphatase. AB - PURPOSE: To compare the efficacy of two tests, alkaline phosphatase (AP) and skeletal alkaline phosphatase (SAP) as staging markers to discriminate patients with cancer of the prostate (CaP) with bone metastases (M+) from those without bone metastases (Mo). MATERIALS AND METHODS: Patients with previously untreated CaP were entered in the retrospective analysis. Serum concentrations of AP (n = 215) and SAP (n = 73) were available. After staging the patients could be divided into 2 groups: Group I: patients with CaP and bone metastases (cT2-4 NxMoss AP: n = 40; SAP: n = 21) Group II: patients with CaP without bone metastases (cT3-4 Nx Mo; pT1-3 No Mo; AP: n = 175; SAP: n = 52). RESULTS: None of the Mo patients but 71% of the M+ patients exhibited a SAP value above the reference range (< 19 ng/ml). This difference is statistically significant (p < 0.001) and resulted in a sensitivity and specificity of 71% and 100%, respectively. The Youden-index is 0.7. In contrast 7% of the Mo patients and only 13% of the M+ patients exhibited a AP value above the reference range (< 170 U/l). This difference is statistically not significant (p = 0.71) and resulted in a sensitivity and specificity of 13% and 93%, respectively. The Youden-index is 0.06. CONCLUSION: SAP could become a useful marker in the evaluation of patients with newly diagnosed CaP as it provides more information than AP concerning the skeletal status of these patients. PMID- 10470214 TI - Serum concentrations of transforming growth factor-beta 1 in patients with benign and malignant prostatic diseases. AB - INTRODUCTION: Transforming growth factor-beta 1 (TGF-beta 1) is found to be most markedly elevated in prostate cancer tissue. We determined the concentration of serum TGF-beta 1 in patients with cancer of the prostate (CaP), and correlated them to tumour stage and serum PSA. METHODS: Serum TFG-beta 1 levels and serum PSA levels were determined in 80 patients with untreated CaP (Group I: pT1 3pNoMo: n = 58; Group II: T1-3pN + Mo: n = 6; Group III: T1-4NxM+: n = 16). Patients with histologically confirmed BPH (n = 50) served as controls. RESULTS: Median TGF-beta 1 levels were not different in CaP- and in BPH-patients (32.86 ng/ml and 35.15 ng/ml respectively; p > 0.05). Furthermore there was no increase in TGF-beta 1 concentrations with advancing tumour stage (Group I: 32.86 ng/ml; Group II: 34.3 ng/ml; Group III: 33.44 (ng/ml; p > 0.05). No correlation with PSA levels could be established (Spearman r = -0.19; p = 0.05). CONCLUSION: TGF-beta 1 was found to be non-discriminative between BPH and CaP. Furthermore no TGF-beta 1 increase could be observed with advancing tumour stage in patients with CaP. PMID- 10470215 TI - Local intratumor immunotherapy of prostate cancer with interleukin-2 reduces tumor growth. AB - BACKGROUND: This study was designed to determine the effectiveness and toxicity of local continuous immunotherapy for prostatic cancer. METHODS: 60 juvenile male Copenhagen rats with Dunning adenocarcinoma of the prostate, implanted subcutaneously into both flanks after proven tumor growth, were treated with either human interleukin-2 (IL-2) depot preparations (n = 30) or albumin (placebo) depot preparations (n = 30) implanted directly next to tumor site. IL-2 depots released IL-2 reliably for more than 24 days. Rat serum was tested during treatment for human IL-2, possibly absorbed from depots, and for rat interferon gamma. RESULTS: IL-2 treatment reduced tumor growth significantly (p < 0.001) compared with albumin treated sites or untreated contralateral sites. No toxicity was observed during treatment. That neither human IL-2 nor rat interferon gamma was detected in serum indicates an exclusively local IL-2 effect. CONCLUSIONS: IL 2 depot preparations reduce tumor growth in Dunning adenocarcinoma of the prostate significantly without toxicity. PMID- 10470216 TI - CYFRA 21-1 in the early diagnosis of recurrent disease in non small cell lung carcinomas (NSCLC). AB - Due to its high specificity and sensitivity CYFRA 21-1 was found to be the leading marker in NSCLC. We focused our interest on the diagnostic value of CYFRA 21-1 in the detection of recurrent disease of 86 patients suffering from NSCLC following R0-resection (median follow up: 22.7 months). Preoperatively, CYFRA 21 1 was positive (cut off 3.3 ng/mL) in 38 of the 86 patients (45%). 48 hours after surgery all 38 patients had CYFRA 21-1-concentrations within the reference range corresponding to a R0-resection. During further follow up 22 of these patients developed local recurrence and/or distant metastases. All 22 patients showed elevated CYFRA 21-1-values at time of detection of relapse, in 8 patients the CYFRA 21-1-increase preceded the detection of recurrence by 2 to 15 months. 16 patients remained disease free and had stable low CYFRA 21-1-values all the time. Out of the 48 preoperatively CYFRA 21-1-negative patients 15 developed recurrent disease. 7 of the 15 patients proved to express cytokeratin 19-fragments at this time. CONCLUSION: CYFRA 21-1 possesses a high specificity and sensitivity in the detection of recurrent disease of patients suffering from NSCLC and with elevated values at time of primary diagnosis. Thus CYFRA 21-1 could contribute to an economical follow up care. Even if there is not the possibility of curative therapy at time of relapse the early use of systemic therapy could be considered. PMID- 10470217 TI - CYFRA 21-1 in the follow-up of inoperable non-small cell lung cancer patients treated with chemotherapy. AB - The introduction of new regimens in the chemotherapy of inoperable non-small cell lung cancer (NSCLC) patients provides a useful extension of survival probability that may now justify the application of tumor markers for the disease monitoring. In a prospective study of 48 consecutive NSCLC patients with TNM stages IIIB/IV we compared changes in the serum levels of the cytokeratin 19 fragment CYFRA 21-1 with the clinical evaluations of response to therapy. CYFRA 21-1 levels were measured using the enzyme immunoassay of Boehringer, Mannheim (Germany). Clinical response to therapy was evaluated according to standard criteria of the WHO. For the assessment of response to therapy by changes in the marker levels the difference between two consecutive levels must exceed 30%. This value is based on the formula: Difference = 2 square root of 2 x CV (CV: inter-assay coefficient of variation of the marker test). CYFRA 21-1 was found to be elevated in 29/48 (60.4%) patients prior to therapy and in 10/48 (20.8%) patients at tumor progression. 91 evaluations have been recorded in these 39 patients. The overall concordance between changes in the marker levels and the clinical assessment was 59.3%. The decrease of CYFRA 21-1 levels at remission was rather low resulting in a concordance of only 42.9%, i.e. marker assays cannot replace the clinical restaging by imaging modalities. In contrast, changes in the marker levels at progression did exceed the required 30% in the majority of cases (64.7%). Most of discordant results (40.7%) could be explained by insufficient decrease or increase of CYFRA 21-1 levels or by extended lead-time. The most striking result was the detection of progressive disease by rising marker levels. Except one case, there was no false-positive elevation of CYFRA 21-1 levels. It is concluded that the detection of progressive disease by rising CYFRA 21-1 levels may avoid continuation of ineffective treatment. PMID- 10470218 TI - Pro-gastrin-releasing peptide (ProGRP)--a useful marker in small cell lung carcinomas. AB - Gastrin-releasing-peptide (GRP), the mammalian counterpart of amphibian bombesin, has been reported to be produced by cells of SCLC. Using recombinant ProGRP Yamaguchi et al developed an enzyme immunoassay for the measurement of this more stable precursor of GRP. We focused our interest on the comparability of ProGRP to neuron specific enolase (NSE), CYFRA 21-1 and CEA. For this purpose we investigated the sera of 272 patients with histologically proven carcinomas of the lung (87 SCLC, 185 NSCLC). The sera of 74 patients with benign diseases of the lung and smokers served as a reference group. At a specificity of 95% ProGRP and NSE possessed comparable sensitivities (47% versus 45%) in small cell lung carcinomas. ProGRP showed only a few more positive test results than NSE, but reached much higher value levels than NSE. ProGRP and NSE showed a clear additive sensitivity of about 20%. In NSCLC CYFRA 21-1 was the leading marker with 63% sensitivity, whereas ProGRP seldom showed a "false positive" test result. ProGRP proved a very high specificity and good sensitivity for small cell lung carcinomas and therefore enables diagnosis of small cell lung carcinoma in patients with lung tumours of unknown origin as well as good control of efficiency of therapy. PMID- 10470219 TI - Treatment of pulmonary metastatic renal-cell carcinoma in 116 patients using inhaled interleukin-2 (IL-2). AB - BACKGROUND: We report 6 years of experience in 116 patients who used inhaled interleukin-2 (IL-2) and were treated in different protocols with natural, recombinant glycosylated and recombinant nonglycosylated IL-2. MATERIALS AND METHODS: All protocols had in common high-dose inhalation of IL-2, either exclusively (11%), with low-dose systemic IL-2 (33%), or with low-dose systemic IL-2 and interferon-alpha (56%). Maximal toxicity per total treatment time (median treatment time, 7.2 months) was mild and at a low incidence (16%) of WHO grade 3 toxicity. Treatment was allowed in patients for whom systemic IL-2 was not suitable. RESULTS: Progressive pulmonary metastases responded in 15% of patients for a median of 15.5 months (range 4.1-33) and were stabilized in 55% for a median of 6.6 months (range, 3-51.7). Overall response rate was 16%, 49%, and 35%, respectively. Median overall response duration was 9.6 mo. Median achieved survival was 11.8 months (range 1.7-68.8). CONCLUSIONS: Inhaled IL-2 prevents progress of pulmonary metastases effectively in 70% of patients. Local use of IL-2 allows the use of the full potential of cytokines with little or no toxicity. PMID- 10470220 TI - Diagnostic accuracy and predictive value of the tumor-associated antigen S100 in malignant melanomas: validation by whole body FDG-PET and conventional diagnostics. AB - We investigated the predictive value of the protein S100 as a tumor-marker in the post-surgical follow-up staging of patients with High Risk melanomas (Clark Levels IV/V, Thickness > 0.75 mm). METHODS: In 51 follow-up studies, performed in 50 patients, serum concentrations of the protein S100 were measured and evaluated with respect to their diagnostic value based on the findings of whole body F-18 FDG-PET studies. The findings of FDG-PET were correlated with the findings of CT, MRI, lymph node sonography, bone scintigraphy, and with histological findings, or a follow-up staging after at least 6 months, respectively. Thus, FDG-PET exhibited a sensitivity of 100% and a specificity of 95%. S100 concentrations were measured by an established RIA (Ria-mat Sangtec 100) and by a novel LIA (Lia mat Sangtec 100 beta). Subsequently, S100 was compared with thymidine kinase for the predictive values of both tumor-markers in the follow-up of malignant melanoma. RESULTS: Intraindividual comparison of S100 concentrations measured by LIA with those obtained by RIA showed a calculated correlation coefficient of 0.97 (Cutoff-levels were 0.1 microgram/l [RIA] and 0.2 microgram/l [LIA]). The arrangement of these cutoff-levels leads to a sensitivity of 85% at a specificity of 55% for Protein S100 when measured by RIA, and to a sensitivity of 77% at a specificity of 61% when measured by LIA. The negative predictive values are 91% (RIA) and 88% (LIA), in association with positive predictive values of 39% (RIA) and 40% (LIA). At a cutoff-level of 4.0 micrograms/l, thymidine kinase showed a sensitivity of 70% and a specificity of 41%. CONCLUSIONS: a) The results obtained by LIA are in a very good correlation with the results measured by the well established RIA. Thus, the LIA is a suitable method for the clinical routine. b) The protein S100 as a tumor-marker of malignant High Risk melanomas has no clinically important sensitivity and specificity as could be proved by a comparison with the predictive value of FDG-PET. c) Whether or not during therapy information concerning progression/remission of the disease can be received out of repetitive measurements of the S100, has to be shown by the results of validated long term follow-up studies. PMID- 10470221 TI - MIA, a novel serum marker for progression of malignant melanoma. AB - MIA was isolated previously as a small soluble protein secreted from malignant melanoma cell lines in vitro. Highly restricted expression patterns in melanocytic tumors were identified in vivo. We therefore quantitated serum levels of MIA protein by means of a non-radioactive ELISA and investigated whether MIA provides clinically relevant parameters in patients with malignant melanomas. Here we report enhanced MIA serum levels in 13% and 23% of patients with stage I and II disease, respectively, and in 100% with stage III or IV disease. Response to therapy in stage IV disease correlated with changes in MIA serum levels and surgical removal of metastases led to normalization of serum values. Repeated measuring of sera from 350 patients with a history of stage I or II melanoma during follow-up, we detected 32 patients developing positive MIA values. At the time of serum analysis 15 of them had developed metastases and one presented with metastatic disease 6 months later. In conclusion, MIA represents a novel serum marker for systemic malignant melanoma revealing the highest sensitivity and specificity among currently available markers. PMID- 10470222 TI - Interstitial high dose rate brachytherapy in locally progressive or recurrent head and neck cancer. AB - In a retrospective study, 19 patients with progressive or recurrent head and neck cancer which had been treated with interstitial high dose rate brachytherapy were analysed. All of them had been previously treated with external radiation. Initial therapy further included surgery in 9 cases and chemotherapy in 3 patients. Staging according to the TNM system revealed advanced stage tumors in the majority of patients. Interstitial brachytherapy was carried out with the isotope Iridium-192. The applied total dose at the reference isodose varied between 10 and 30 Gy. Application was fractionated once a week. A complete tumor remission was achieved in 5 patients and partial remission in 10 patients. In 4 patients the tumor continued to grow despite brachytherapy. The mean follow-up in our collective was 21 months. The calculated local control rate was 34% at 24 months. The survival rate was 49% at 12 months and 35% at 24 months. Interstitial brachytherapy is recommended as a palliative treatment in preirradiated squamous cell carcinoma with local recurrence or progression. PMID- 10470223 TI - CYFRA 8/18 in head and neck cancer. AB - BACKGROUND: Cytokeratins (Ck) 8, 18 and 19 that are normally expressed in head and neck tissue in only small amounts, become overexpressed in head and neck squamous cell cancerous tissue. It was questioned whether Ck 8 and 18, which occur together, were also detectable and of value as a tumor markers for this cancer entity. MATERIALS AND METHODS: One hundred forty-nine sera from patients with squamous cell carcinoma of the head and neck (SCCHN) were investigated with a novel ELISA kit for the detection of fragments of Ck 8 and 18. Twenty-five sera from healthy volunteers and 39 patients with benign diseases of the head and neck region served as controls. RESULTS: It was found that CYFRA 8/18 had a sensitivity of 7%. CYFRA 8/18 values did not show a correlation with clinical parameters. CONCLUSIONS: Because of a low sensitivity CYFRA 8/18 seems not to be of value as a tumor marker for SCCHN. PMID- 10470224 TI - A new prognostic indicator for head and neck cancer--p53 serum antibodies? AB - BACKGROUND: p53 antibodies are a new serological parameter of unknown potential in patients with malignancies. Their occurrence has been described in various types of cancer patients and several studies in head and neck cancer patients and other cancer patient groups have indicated its prognostic value. MATERIAL AND METHODS: We investigated the incidence of p53 serum antibodies in 271 head and neck cancer patients with an ELISA and investigated a possible correlation with clinical parameters like tumor staging and grading. RESULTS: Sixty-seven head and neck cancer patients (24.7%) were seropositive for p53 antibodies. No correlation between p53 antibody status, tumor staging and grading was found. CONCLUSIONS: These results indicate that the occurrence of p53 antibodies does not correlate with the most relevant prognostic factors, rate of regional metastasis and primary tumor size, in patients with head and neck cancer. PMID- 10470225 TI - The influence of differing dissection techniques on p53 as a possible indicator for local recurrences in carcinomas of the upper aerodigestive tract. AB - BACKGROUND: A correlation between the occurrence of p53 protein in margins after tumor resection has been proposed as indicator for recurrence. This study should clarify whether differences occur in p53 protein detection after laser surgery as opposed to surgery with the scalpel. MATERIAL AND METHODS: Healthy mucosa of the oropharynx (n = 28) and squamous cell carcinoma of the head and neck (SCCHN) (n = 19) were excised with scalpel, CO2 and Nd:YAG laser and then examined for the presence of p53 protein with immunohistochemistry and ELISA. RESULTS: The occurrence of p53 protein in SCCHN was dependent on the applied resection technique. In lasersurgically excised tissue only a small zone adjacent to the cutting edge was found to be negative for p53. This zone of necrosis was smaller in margins after CO2 than after Nd:YAG laser resection. CONCLUSIONS: Lasersurgical excision does only inhibit the detection of p53 in the zone of necrosis. PMID- 10470226 TI - Thyroid peroxidase (TPO) as a tumor marker in the follow-up of differentiated thyroid carcinomas with surgical and ablative radioiodine therapy. An assessment after evaluation. AB - The clinical significance of serum thyroid peroxidase (TPO) for differentiated thyroid carcinomas(DTA) is estimated differently. In our preliminary studies it was found that TPO presented information extending those that from hTG. For further clarification a prospective follow-up study was performed including 66 female and 14 male total thyroidectomized patients with DTA for the time course of TPO and human thyroglobulin (hTg) in relation to the ablative radioidine therapy (ART). In 34/50 evaluable cases TPO levels showed an approximately analogous time course with hTg. In relation to the extension of residues, some cases presented increasing of TPO and hTG after radioiodine treatment. 6/7 patients suffering from extended postoperative residues with high anti hTg levels but without elevated TPO concentrations showed distinctly elevated TPO values. Therefore, TPO seems to be an indicator for the destruction of normal thyroid cells or thyroid tumor cells. The clinical value of TPO seems to be in the time being limited to DTA due to false negative hTg values. However, it should be possible that TPO can did the evaluation of thyroid specific therapy. PMID- 10470227 TI - Persistent elevation of CA 19-9 levels in a patient with an extended retroperitoneal dermoid. AB - CA 19-9 has been established as sensitive tumor marker in a variety of malignant diseases, especially in carcinomas of the exocrine pancreas. The specificity of CA 19-9 is limited by many benign diseases causing abnormal values. A case of a patient with a retroperitoneal dermoid showing high serum levels of CA 19-9 will be described. The patient suffered from a large retroperitoneal mass (7 x 6 x 9 cm). A CT-guided fine needle biopsy of the tumor was performed and the histology was compatible with a benign dermoid. The CT-scans of the abdomen showed also the typical features of a dermoid. Tumor markers and lab counts stayed in the normal range except CA 19-9, which was constantly elevated with value between 131 and 329 U/ml. A benign or malignant disorder was excluded. An immunoscintigraphy and a SPECT with a I-131-labeled monoclonal antibody against CA 19-9 revealed the retroperitoneal mass as the source of the CA 19-9 elevation. This is the first case described in English scientific literature of a retroperitoneal dermoid (benign teratoma) as a source of a persistently elevated level of CA 19-9. PMID- 10470228 TI - Apoptosis in serum of patients with solid tumours. AB - Apoptosis, which occurs in highly proliferating tumours spontaneously or during anticancer therapy, many lead to an elevated concentration of circulating nucleosomes in blood. In order to quantify the concentration of nucleosomes, we used the Cell Death Detectionplus-ELISA (CDDE) (Boehringer Mannheim, Germany) based on antibodies against histone and DNA, adapted it to the demands of liquid materials and standardized test performance and handling of serum. Furthermore, we investigated serum samples of 185 patients with solid tumours (additionally 24, treated with radio- or chemotherapy), 30 with acute inflammations and 50 healthy persons. Many patients with tumours (78%) and inflammations (77%) showed higher concentrations of serum-nucleosomes (> 100 AU), whereas 96% of all healthy persons had low values (< 100 AU). Follow up-studies revealed an early peak after initiation of therapy and correlated to the clinical outcome. The concentration of nucleosomes is a sensitive marker of cell death and could be used for monitoring the efficacy of antitumour therapy. PMID- 10470229 TI - Rat tumors following fractionated irradiation. AB - BACKGROUND: Rats are susceptible to irradiation and can develop benign and malignant tumors either spontaneously or in the field of irradiation. In the head and neck region, there are no reports available on the type of tumor after fractionated irradiation using a human therapy protocol. MATERIAL AND METHODS: We analyzed 19 tumors, in 19 rats, which developed after external X-irradiation of the left neck area in Wistar rats (2 Gy/day, monofractions, 5 days/week, total dosage 60 Gy) and compared the findings with tumors in untreated rats of the same strain. RESULTS: Tumors in the irradiation field proved to be squamous cell carcinoma or adenoid cystic carcinoma (ACC), not sarcoma. These entities were sporadically found in non-irradiated rats at a higher age. CONCLUSIONS: ACC has rarely been reported in the literature on laboratory rats. The development of this highly aggressive malignant tumor can be expected 3 months to 1 year after completion of irradiation. PMID- 10470230 TI - Multicenter evaluation of the Elecsys CA 125 II assay. AB - The technical performance and clinical usefulness of the newly developed Elecsys CA 125 II assay (Boehringer Mannheim) was evaluated in a multicenter study. Imprecision studies were carried out using control sera and human pool sera with CA 125 concentrations from 11 to 1026 U/ml. Within-run CVs between 0.7 to 4.8% (median 1.7%) and between-day CVs between 2.4 to 10.9% (median 5.7%) were found. Method comparison studies with Enzymun-Test CA 125 II carried out in four laboratories yielded slopes between 0.94 to 1.07 and intercepts < 3 U/ml. A good comparability of the Elecsys CA 125 II assay was also found with one MEIA and the Centocor" IRMA. For a second MEIA and a second IRMA the slopes were 1.23 and 1.42, and the corresponding correlation coefficients were 0.987 and 0.977, respectively. The Elecys CA 125 II concentrations are clearly related to the tumor stage of ovarian carcinoma patients. The maximum of diagnostic efficiency of ovarian carcinoma patients compared with patients of benign gynecological diseases is reached at 150 U/ml with a specificity of 93% and a sensitivity of 69%. Follow-up studies of ovarian carcinoma patients reflect the status of the disease and the effect of various therapeutic applications. The technical and clinical evaluation of the Elecsys CA 125 II assay show a superior analytical performance with a broad measuring range up to 5000 U/ml and a short measuring time of 18 minutes. PMID- 10470231 TI - LIAISON hCG--an automated chemiluminescent immunoassay for the determination of human chorionic gonadotropin (hCG). AB - An immunoassay utilizing chemiluminescence and paramagnetic particles has been developed for the new fully automated, random access LIAISON immunoanalyzer. The LIAISON hCG assay is a two-site immunolouminometric one-step assay using two highly specific monoclonal antibodies. Total incubation time is only 10 min. A specially designed unique Reagent Integral contains all specific reagents; the on board stability of these reagents is given over a very long period (> 4 weeks). The assay works with a 2-point calibrated master curve. 30 microliters sample is added to 200 microliters tracer and 20 microliters antibody-coated magnetic particles. After 10 min incubation the particles are separated, washed and the chemiluminescent signal is generated. The time to first result is only 15 min. The assay with a unique extended standard range up to 5,000 ng/ml shows no high dose hook effect up to 500,000 mlU/ml (spiked sera). The assay detects both the intact molecule and the free beta subunit. The cross-reactivity to FSH, LH and TSH is less than 0.1%. Precision (within-run < 3%; between-run < 5%), linearity, recovery and sensitivity (< 0.5 mlU/ml) are excellent. The assay shows a very good correlation to the LIA-mat hCG (r = 0.996). In summary the LIAISON hCG assay together with the new LIAISON immunoanalyzer is a very rapid and accurate method for the quantitative determination of hCG/beta-hCG in serum. PMID- 10470232 TI - LIAISON Ferritin--an automated chemiluminescent immunoassay for the determination of Ferritin. AB - An immunoassay utilizing chemiluminescence and paramagnetic particles has been developed for the new fully automated random access LIAISON immunoanalyzer. The LIAISON Ferritin assay is a two-site immunoluminometric one-step assay using two highly specific monoclonal antibodies. A specially designed unique Reagent Integral contains all specific reagents. The cooled reagent compartment guarantees an on-board stability of at least 4 weeks. The assay works with a 2 point calibrated master curve. To perform, the assay, 10 microliters sample is added to 300 microliters tracer and 20 microliters antibody-coated magnetic particles. After 10 min incubation the particles are separated, washed and the chemiluminescent signal is generated. The time to first result is only 15 min. The assay with a unique extended standard range up to 3,000 ng/ml shows no high dose hook effect up to 150,000 ng/ml (spiked sera). The cross-reactivity to human liver ferritin is 100%, to human spleen ferritin 66% and to human heart ferritin 8%. Precision (within-run < 3%; between-run < 5%), linearity, recovery and sensitivity (< 0.2 ng/ml) are excellent. The assay shows a very good correlation to LlA-mat Ferritin (r = 0.994). In summary the LIAISON Ferritin run on the new LIAISON immunoanalyzer and is a very rapid and accurate method rapidly providing reliable results. PMID- 10470233 TI - Performance characteristics of IMMULITE TPS: a comparison with TPS IRMA. AB - This paper describes an automated immunoassay for the measurement of TPS developed for the IMMULITE. The IMMULITE system is a fully automated continous random access analyzer, which uses enzyme-amplified chemiluminiscent as the detection system. The IMMULITE TPS assay is a sequential, two-site chemiluminiscent enzyme immunometric assay designed for the quantitative measurement of TPS in serum. The IMMULITE TPS assay covers a clinical concentration range of 20-2400 U/L, with a lower detection limit of 6 U/L. A serum method comparison (n = 340) to the TPS IRMA assay demonstrates a correlation coefficient of 0.961 and a slope of 1.04. Samples run on IMMULITE TPS assay exhibit linearity upon dilution where recovered values fall within 15% of the theoretical sample value. The intra-assay precision of the IMMULITE TPS assay ranged from 3.3-4.7%, while the interassay precision ranged from 3.9-6.0%. By using different methods (immunoradiometric assay vs chemiluminiscent enzyme labeled immunometric assay, one-step procedure vs sequential two-step procedure, manual vs fully automated procedure) the two TPS tumor marker tests used in this study gave quite comparable determinations for the sera from the cancer patients. PMID- 10470234 TI - Multicentre tumour marker reference range study. AB - BACKGROUND: The array of antibodies and assay formats utilised by kit manufacturers contribute to different values being quoted as clinically "normal". Kit-specific reference range limits will maximise the clinical utility of tumour marker assays. MATERIALS AND METHODS: Serum samples (approximately 800) were obtained from volunteers, age 20-70 years, in France, Germany, The Netherlands and Portugal. Each sample was assayed with a number of DPC tumour markers kits. IMMULITE assays were carried out in The Netherlands; Coat-A-Count IRMA, IRMA Count, Double Antibody and Milenia assays at EURO/DPC Ltd. Analytes included, BR MA, OM-MA, GI-MA (for the determination of CA 15-3, CA 125 and CA 19-9, respectively), CEA, PSA, PAP and HCG. RESULTS: The median and 95th percentiles for each analyte in each assay format were estimated; where appropriate, data subsets were considered. CONCLUSION: Kit-specific reference range data generated for important tumour marker analytes will help clinicians interprete their assay results. PMID- 10470235 TI - Quantitative detection of tumor M2-PK in serum and plasma. AB - The majority of tumors express an isoform of the glycolytic enzyme pyruvate kinase, the type tumor M2. The isoenzyme exists in an active tetrameric and a less active dimeric form. The dimeric form is strongly overexpressed in tumor cells and this new tumor marker is thus called tumor M2-PK. This isoenzyme is released from tumor cells and is quantitatively detectable in body fluids by a sensitive enzyme-linked immunosorbent assay (ELISA). To establish the ELISA for the routine diagnostic in a clinical laboratory, the sample stability was evaluated. Therefore, blood samples were collected in different ways from healthy donors. Reproducibility of tumor M2-PK detection in EDTA-plasma was not affected by the day to day 'stress' in a clinical routine (e.g. shaking, leaving the samples at room temperature for several hours without prior centrifugation). Similar results were obtained with citrate-plasma, whereas detection in serum and heparin-plasma was only reproducible when the blood samples were centrifuged within 2 hrs after collection. It appears that lymphocytes contain small amounts of the tumor M2-PK isoenzyme. They might release tumor M2-PK in heparin-plasma and serum samples, but not in EDTA-plasma samples. The results indicated that EDTA-plasma appears to be most appropriate for the routine diagnostic of tumor M2 PK as a tumor marker. PMID- 10470236 TI - Development of a new automated enzyme immunoassay for the determination of neuron specific enolase. AB - Neuron-specific enolase (NSE) represents the gamma gamma- and alpha gamma- isoforms of the dimeric glycolytic enzyme enolase. NSE is predominantly found in neurons and neuroendocrine cells and has proven to be a marker for tumors derived from these cells. It is widely accepted in the monitoring of patients with small cell lung cancer and is also of value as an aid in diagnosis. Recently it has become of interest in the monitoring of brain damage. Monoclonal antibodies against gamma-enolase were raised in mice and selected for optimal performance on the Cobas Core enzyme immunoassay system. The antibody combination of choice was MAb 18E5 for capturing and MAb 84B10 for detection which is accomplished by using a horseradish peroxidase conjugate and the substrate 3,3',5,5' tetramethylbenzidine. The resulting assay is a one-step enzyme immunoassay of the sandwich type. It is performed on the fully automated Cobas Core immunoassay analyzer with a total assay time of 45 min. The sample volume is 10 microliters. Calibration is done by a 1-point recalibration using a lot-specific master calibration curve provided with the kit. The dynamic range is 0-200 ng/ml. The analytical detection limit (standard 0 + 2SD) of the Cobas Core NSE EIA II was 0.1 ng/ml. Intra- and interassay coefficients of variation were < 5% and < 6%, respectively. A Hook Effect was not observed up to a concentration of 20'000 ng/ml. Test results correlated closely with the well established polyclonal Cobas Core NSE EIA (r = 0.99). In summary, the Cobas Core NSE EIA II is a rapid, reliable and convenient test for measuring NSE in human serum. PMID- 10470237 TI - Liaison free PSA--an automated chemiluminescent immunoassay for the determination of free prostate specific antigen (fPSA). AB - The LIAISON fPSA offers a rapid, reliable and precise method for the fully automated determination of free PSA. PMID- 10470238 TI - Automated Cobas Core PSA assays: sensitive, precise, and specific measurement of PSA-total and PSA-total/PSA-free ratio. AB - We describe the analytical performance of the newly developed Roche Cobas Core PSA Total EIA and the Cobas Core PSA Free EIA. The assays are designed for the random access immunochemistry analyzer Cobas Core. Both assays are highly precise (CV's < 4%) and highly sensitive (PSA Total: analytical sensitivity < 0.02 ng/ml; PSA Free: < 0.01 ng/ml). The PSA Total EIA measures PSA-ACT and PSA free equimolarly. The assays are standardized against the reference material of the Stanford group and correlate very well with the designated golden standard Hybritech Tandem-R assays. The high-dose-hook is beyond 20,000 ng/ml and 4500 ng/ml for the PSA Total and PSA Free assay, respectively. In summary, the data indicate clearly the reliability of the measuring system and the achievement of the demands of PSA diagnostics. PMID- 10470239 TI - Disseminated tumor cells versus micrometastasis: definitions and problems. AB - The finding of single tumor cells or small tumor cell groups in the lymph and blood (including bone marrow aspirations) should be designated as isolated or disseminated tumor cells. The UICC (1992) recommended to identify such findings by the addition of "(i)" (for isolated tumor cells). In contrast, micrometastasis (2 mm or less) should be indicated by the addition of "(mi)". Before introduction of the examination of lymph nodes or bone marrow aspirations regarding isolated (disseminated) tumor cells the methods have to be standardized and the clinical relevance (independent prognostic significance, assistance in selection of patients for adjuvant treatment) has to be evaluated in prospective large scale studies. PMID- 10470240 TI - In vitro toxicity assessment of mouthwashes in the pollen tube growth test: first results. AB - Mouthwashes are widespread used to treat halitosis or minor oral infections. Adverse effects of mouthwashes were recognized, e.g. desquamation of the oral mucosa and stomatitis. The acute toxicity of mouthwashes was tested in an in vitro system, the pollen tube growth test (PTG-test). In the PTG-test the acute toxicity of mouthwashes exceeds the toxic effect of alcohol. Consumers should be informed about the number and amounts of mouthwash ingredients, and their toxic potential. These declarations would provide the basis for testing the carcinogenic effects of mouthwashes. PMID- 10470241 TI - Immunohistochemical detection of extracellular matrix proteins in the irradiated rat mandibular gland. AB - BACKGROUND/MATERIAL AND METHODS: In 59 rat mandibular glands we investigated the distribution of extracellular matrix proteins (EMP) in rat salivary gland tumors. The animals differed in age and pretreatment status (irradiation versus no irradiation). For immunohistochemical analysis we used polyclonal EMP antibodies [anti-laminin (AL), anti-fibronectin (AF), anti-collagen-III (AC-III), anti collagen-IV (AC-IV)]. RESULTS: The antibodies identified the different structures of the extracellular matrix: components of basement membranes [(BM); laminin, collagen IV] and components of BM-associated structures (fibronectin, collagen III). Immunoreactivity was found in glandular tissue, in vascular walls, in nerve tissue and the interstitial connective tissue. Various EMP showed different patterns. Significant differences were seen between irradiated and non-irradiated glands, often with generally stronger and more extended staining in the irradiated group. Thus, both nerve tissue and excretory ducts (ECD) were in part strongly stained for AL, while ECD were intensely positive for AF. CONCLUSIONS: Preceding radiation has to be considered when interpreting salivary gland tissue, especially in tumor differentiation studies following irradiation. PMID- 10470242 TI - Quality requirements and control: EGTM recommendations. European Group on Tumour Markers. PMID- 10470243 TI - Tumour markers in germ cell cancer: EGTM recommendations. European Group on Tumour Markers. PMID- 10470244 TI - Tumour markers in prostate cancer: EGTM recommendations. European Group on Tumour Markers. PMID- 10470245 TI - Tumour markers in breast cancer--EGTM recommendations. European Group on Tumor Markers. PMID- 10470246 TI - Tumour markers in gynaecological cancers--EGTM recommendations. European Group on Tumor Markers. PMID- 10470247 TI - Tumour markers in gastrointestinal cancers--EGTM recommendations. European Group on Tumour Markers. PMID- 10470248 TI - Tumour markers in lung cancer: EGTM recommendations. European Group on Tumour Markers. PMID- 10470249 TI - Internal initiation of translation efficiency in different hepatitis C genotypes isolated from interferon treated patients. AB - Initiation of translation of hepatitis C viral RNA occurs internally and it is mediated by a segment of about 330 nucleotides termed Internal Ribosome Entry Site (IRES) located in the 5' end region. While being the most conserved part of the genome, this region also accumulates nucleotide substitutions which are often covariant. In this study we have examined the activity and sequence variation of IRES elements belonging to genotypes 1b, 2a/2c and 3a in patients that responded or not to interferon therapy. The substitutions found in the IRES region analyzed were predicted to maintain the secondary structure of the RNA. Comparison of their efficiency to promote internal initiation of translation in bicistronic constructs supported the conclusion that for both 1b and 3a genotypes, response to interferon therapy and IRES activity are unrelated, although sequence homology was not always found among isolates from patients with different type of response. IRES activity of the studied genotypes varied about 4-fold under the conditions used in our in vivo assays depending on the cell line used for transfection. Such differences were not evidenced in vitro suggesting that the differences observed depend on trans-acting factors present in the transfected cell. PMID- 10470250 TI - Complete nucleotide sequence of the genomic RNA of a Japanese yam mosaic virus, a new potyvirus in Japan. AB - We determined the complete nucleotide sequence of a potyvirus purified from a Japanese yam plant. The genomic RNA of this virus is 9,757 nucleotides (nts) in length, excluding the 3'-terminal poly(A) tail. It contains a single open reading frame (ORF) encoding a polyprotein of 3130 amino acids (aa) with a calculated Mr of 356,793. The genomic organization of this potyvirus is similar to that of other members of the genus Potyvirus and nine potential cleavage sites for the viral proteinase were found by comparison of its sequence with those available for other potyviruses. The nucleotide sequence and genome characteristics show that this isolate is a new potyvirus species. Its polyprotein differs substantially from Yam mosaic virus (YMV) (50% amino acid sequence identity) and fourteen other potyvirus species examined (44-59% identity). Although this potyvirus has been classified as YMV, our results suggest that the potyvirus infectious to the Japanese yam plant in Japan is distinct from YMV. Therefore, we propose that the Japanese yam potyvirus should be designated as Japanese yam mosaic virus (JYMV). PMID- 10470251 TI - Tumorigenicity of mouse BALB/c 3T3 fibroblast cells which express Epstein-Barr virus-encoded LMP1 and show normal growth phenotypes in vitro is correlated with loss of transforming growth factor-beta 1-mediated growth inhibition. AB - Latent membrane protein 1 (LMP1) encoded by the Epstein-Barr virus (EBV) genome is known to induce loss of contact inhibition and the anchorage-independent growth in rodent fibroblasts and increased expression of cell-surface activation markers and cell adhesion molecules in human B lymphocytes. To analyze the role of LMP1 in tumorigenicity, we prepared BALB/c 3T3 clones (B3LP) expressing LMP1. These B3LP cells showed non-transformed phenotypes in vitro which were characterized by normal cell morphology, contact inhibition in growth and anchorage-dependent growth. The activity of NF-kappa B induced generally in several cell lines after transfer of the LMP1 gene was not detected in B3LP cells. However, B3LP expressing LMP1 at moderate levels lost sensitivity to growth arrest by transforming growth factor-beta 1 (TGF-beta 1) and formed tumors in severe combined immune deficiency mice. Cells expressing the truncated form of LMP1 and expressing LMP1 at low level were sensitive to TGF-beta 1-mediated growth arrest and did not form tumors in mice. Therefore, cells expressing LMP1 at moderate but not at low levels formed tumors in mice and lost sensitivity to TGF-beta 1. Our results suggest that loss of TGF-beta 1-mediated growth inhibition is an important event for the tumorigenicity of LMP1-expressing cells. PMID- 10470252 TI - In situ localization of cacao swollen shoot virus in agroinfected Theobroma cacao. AB - Cacao swollen shoot virus (CSSV) is a small non-enveloped bacilliform virus with a double-stranded DNA genome. A very restricted host range and difficulties in transmitting the virus, either mechanically or via its natural vector, have hindered the study of cacao swollen shoot disease. As an alternative to the particle-bombardment method previously reported, we investigated another approach to infect Theobroma cacao. A greater-than-unit length copy (1.2) of the CSSV DNA genome was cloned into the Agrobacterium binary vector pBin 19 and was transferred into young plants via Agrobacterium tumefaciens. Typical leaf symptoms and stem swelling were observed seven and eleven weeks post inoculation, respectively. Viral DNA, CSSV coat protein and virions were detected in leaves with symptoms. Agroinfected plants were used to study the in situ localization of CSSV and its histopathologic effects in planta. In both leaves and petioles, virions were only seen in the cytoplasm of phloem companion cells and of a few xylem parenchyma cells. Light microscopy showed that stem swelling results from a proliferation of the xylem, phloem and cortex cells. PMID- 10470253 TI - Feline peripheral blood mononuclear cells express message for both CXC and CC type chemokine receptors. AB - The feline immunodeficiency virus (FIV) causes a disease in cats similar in clinical presentation and disease progression to that of HIV and AIDS. It is now known that, for HIV infection, as well as primary binding of virus to the CD4 receptor, entry and infection of cells requires coreceptors which are members of the chemokine group of G-protein coupled receptors. Because of the similarity of HIV and FIV, we hypothesised that coreceptors are required for the entry and infection of cells by FIV. Using a feline cDNA library derived from a feline IL-2 sensitive lymphocyte cell line, we identified the presence of message for both CC and CXC chemokine receptors. The feline CXCR4 has been shown to facilitate fusion by FIV [44] and we suggest that the feline CCR5 receptor mediates infection of feline cells by M-tropic strains of FIV. PMID- 10470254 TI - Identification, sequence analysis and characterisation of equine herpesvirus 5 glycoprotein B. AB - The complete nucleotide sequence of the gammaherpesvirus equine herpesvirus 5 (EHV5) glycoprotein B (gB) was determined and the deduced amino acid sequence compared with that of the second equine gammaherpesvirus EHV2. EHV5 gB is an 870 amino acid protein and is 79% similar and 66% identical with EHV2 gB at the amino acid level. EHV5 gB like EHV2 gB is a disulphide linked heterodimer with subunits of 92 and 68 kDa. EHV5 gB is an integral membrane glycoprotein containing only N linked oligosaccharides and contains a putative endoproteolytic cleavages site at amino acids 422-485. The EHV5 gB amino acid sequence showed greatest homology with other members of the Rhadinovirus genus of the subfamily Gammaherpesvirinae. Alignment of EHV5 gB sequence with the gB sequence of seven other gammaherpesviruses showed conservation of 10 cysteine residues as well as conservation of three predicted sites of N-linked glycosylation; the highest degree of conservation of the predicted sites of N-linked glycosylation was observed between EHV5 and the other members of the Rhadinovirus genus. Phylogenetic analysis confirmed EHV2 and EHV5 were most closely related to each other and equally distant from other members of the Rhadinovirus genus included in the analysis. PMID- 10470255 TI - Australian isolates of ryegrass mosaic rymovirus and their relationships. AB - The sequences of the 3'-terminal 1.8 kb of the genomes of three Australian and three Welsh isolates of ryegrass mosaic rymovirus (RGMV) were determined, as too were the virion protein genes of two New Zealand isolates of RGMV. They were compared with each other and with the published sequences of a Danish and a South African isolate by distance and maximum likelihood methods, and found to be very closely related (mean nucleotide difference 5.5%). All three Australian isolates and one from North Island of New Zealand formed one consistent cluster, and the Danish and South African isolates formed another. However the relationships between these two clusters and the other isolates were not consistent; they depended on the method of comparison used, and on the protein, gene or codon position compared. Nonetheless the European (Welsh and Danish) sequences were 2-4 times more different from one another than those from the Antipodes, suggesting that the European RGMV population may be older than the Antipodean. The Danish isolate has 39 nucleotides of the 5'-terminal region of its virion protein gene frameshifted -1 relative to the 'common' sequence. Interestingly the South African isolate has a similar frameshift, but sequence comparisons indicate that this frameshift must have occurred independently; a possible example of 'convergent frameshifting'. PMID- 10470257 TI - Subcellular localization of hepatitis C viral proteins in mammalian cells. AB - We determined the subcellular localization of hepatitis C viral (HCV) proteins as a first step towards the understanding of the functions of these proteins in the mammalian cell (CHO-K1). We used fluorescence emitted from green fluorescent protein (GFP)-fused to the viral proteins to determine the subcellular localization of the viral proteins. We found that most of the viral proteins were excluded from the nucleus. Core exhibited a globular pattern near the nucleus. NS2 was concentrated in the perinuclear space. NS4A accumulated in the ER and the Golgi regions. NS3 was detected in the nucleus as well as the cytoplasm, when it was expressed by itself. However, NS3 became restricted to the cytoplasm, when it was produced together with NS4A. NS4B showed a spot-like pattern throughout the cytoplasm. NS5A and NS5B were distributed throughout the cytoplasm in a mesh-like pattern. These results can provide a basis for further investigations into the functions of the HCV proteins. PMID- 10470256 TI - Alteration of the cellular response to interleukin-1 beta by SV40 large T antigen in rheumatoid synovial fibroblasts. AB - The large T antigen of SV40 (LT) has been widely used to immortalize primary cells for various studies. In this study, synovial fibroblasts of a patient from rheumatoid arthritis (RA) were transformed with LT gene to analyze the effect of SV40-mediated transformation on the production of cytokines, such as IL-6, IL-8, and GM-CSF, that are under the control of interleukin-1 beta (IL-1 beta), a physiological inducer of nuclear factor kappa B (NF-kappa B). It was noted that the basal levels of GM-CSF and IL-8 were upregulated, whereas that of IL-6 was downregulated. Moreover, the extents of induction of these cytokines in response to IL-1 beta were markedly downregulated in synovial fibroblasts transformed by LT as compared from parental cells. Although IL-1 beta could translocate NF-kappa B to the nucleus in all cells, some of the transformed cells exhibited nuclear translocation of NF-kappa B even before the stimulation with IL-1 beta, suggesting that transformation of LT resulted in the constitutive activation of NF-kappa B, either directly or indirectly. In order to examine whether LT downregulate the kappa B-dependent gene expression, we performed the transient luciferase gene expression assay. We found that cotransfection of LT did not downregulate the kappa B-dependent gene expression that was stimulated with L-1 beta. These observations suggest that the apparent inhibitory effect of LT on the IL-1-induced expression of cytokines may not be through its direct action on the NF-kappa B transactivation. PMID- 10470258 TI - Analysis on reassortment of rotavirus NSP1 genes lacking coding region for cysteine-rich zinc finger motif. AB - Rotavirus clones A5-10 and A5-16 isolated from a bovine rotavirus strain A5 possess NSP1 gene which has a point mutation generating a nonsense codon and a 500 base-deletion, respectively. As a result, the two A5 clones encode truncated NSP1 product which lacks cysteine-rich region forming zinc finger motif. In order to analyze reassortment of these mutated NSP1 gene with RNA segments from heterologous strains, we investigated a number of reassortant clones derived from coinfection with either A5-10, A5-16 or a reference strain A5-13 (possessing intact NSP1 gene) and either simian rotavirus SA11 or human rotavirus KU. In coinfection with SA11 and A5-13, selection rates of A5-13 segments in reassortants ranged approximately from 20 to 70% (46% for NSP1 gene). In contrast, in the reassortment between SA11 and A5-10 or between SA11 and A5-16, selection rates of NSP1 gene from A5-10 and A5-16 were only 1% (one clone) and 0%, respectively. In reassortants from crosses KU x A5-clones, selection rate of A5-13 NSP1 gene decreased to 15%, while 11 reassortants with A5-10 NSP1 gene (31%) and one reassortant with A5-16 NSP1 gene (2%) were isolated. Reassortants with A5-10 NSP1 possessed a single gene (segment 9 or 11) from KU in the genetic background of A5-10. One reassortant clone (cl-55) with A5-16 NSP1 gene possessed KU gene segments 3, 4, and 8-11. When single-step growth curves were compared, the reassortant cl-55 showed almost identical growth curve to that of KU, while KU showed a better replication than A5-16. These results indicated that although A5-10 or A5-16 NSP1 gene encoding the truncated NSP1 is selected into reassortants much less efficiently than normal NSP1 gene, the reassortants with the mutated NSP1 gene and RNA segments from heterologous strains normally replicated in cultured cells. Thus, cysteine-rich region of NSP1 was not considered essential for genome segment reassortment with heterologous virus. PMID- 10470259 TI - Follow-up of hepatitis C virus RNA in peripheral blood mononuclear cells during interferon therapy. AB - Mononuclear cells can be infected in vitro by hepatitis C virus and the viral RNA can be detected in mononuclear cells of chronically infected patients. It was suggested that the virus could persist in the mononuclear cells of some patients treated by interferon. The aim of this study was to follow the presence of viral RNA in the plasma and peripheral blood mononuclear cells of 16 chronically infected patients treated by alpha 2b interferon for 1 year. The RNA was detected by reverse transcription followed by nested PCR and quantified using the branched DNA method at regular intervals for at least one year. Before PCR, the mononuclear cells were treated by RNase and trypsin in order to eliminate the viral particles that could be stuck at the cell surface. Six patients were non responders and had persistent plasmatic viral RNA during the treatment. Two patients were good responders and had persistently negative PCR in both plasma and mononuclear cells. Eight patients had initial negativation of plasmatic hepatitis C virus RNA but showed a relapse characterized by positive plasmatic PCR. Positive PCR in mononuclear cells despite negativity of plasmatic PCR was noted 18 times in 8 patients. Persistently positive PCR in mononuclear cells in absence of detectable viraemia was followed by a virological relapse in 5 of these patients. This study confirms that hepatitis C virus RNA can be detected in mononuclear cells despite negative plasmatic PCR in patients treated by interferon. Moreover, the persistence of viral RNA in peripheral mononuclear cells could be a predictive factor of treatment failure. Our data also suggest that detection of viral RNA in mononuclear cells is probably not only due to passive virus adsorption from plasma. PMID- 10470260 TI - Recombinants of cucumber mosaic virus (CMV): determinants of host range and symptomatology. AB - 117F and R are subgroup I and II cucumber mosaic virus (CMV) strains, respectively. Whereas I17F induces severe symptoms on all hosts so far tested, R induces generally mild symptoms, except on Nicotiana glutinosa, on which it causes leaf blistering and severe stunting. Pseudorecombinants and recombinants, based on RNAs 1 and 2 from R-CMV, were created by adding either I17F RNA 3 or one of two chimeric RNAs created by exchanging approximate halves of RNA 3 of the two strains. The viruses created were tested on different hosts of the virus. On maize, local necrotic lesions were induced by all strains with the 5' part of RNA 3 from R-CMV, whereas only I17F-CMV induced a systemic infection. The seven solanaceous hosts tested could be classified into two main groups. In the first, RNA 3 was not directly involved in the symptoms that were systemically induced, and the extreme disease severity induced by I17F was correlated with high virus accumulation. In the same hosts, the lesser virulence of R-CMV could reflect a deficiency in long-distance movement, involving RNA 3. The second group included Nicotiana glutinosa where the symptoms induced by R-CMV were determined by the 3' part of RNA 3. PMID- 10470261 TI - Analysis of the latency-associated transcript/UL1-3.5 gene cluster promoter complex of pseudorabies virus. AB - During latency, pseudorabies virus (PRV) DNA is preferentially retained in the neurons of the trigeminal ganglion and a spliced 8.5-kilobase poly-A RNA, designated large latency transcript (LLT), is synthesized. Because LLT is the only transcript made during the latent phase, the LLT promoter may be unique among all other PRV promoters that are active in productive infections. Organization of the PRV LLT promoter is quite complex because it coincides with the UL1-3.5 gene cluster promoter, but in the opposite orientation. By conventional designation, LLT is transcribed in the rightward direction while the UL1-3.5 gene cluster is transcribed in the leftward orientation. In this work, activities of the LLT promoter and the UL1-3.5 gene cluster promoter were investigated by transient reporter gene expression assay in cells of neuronal and non-neuronal origins. There are two TATA boxes in this region. We examined the promoter activities of the first TATA box with its 5' sequence (LAP1) and the second TATA box with its 5' sequence (LAP2). The UL1-3.5 promoter driven constructs gave no reporter gene activity in any of the experiments. Reporter gene activity was detected with LAP2 gene constructs, but not with LAP1 constructs, in both neuronal and non-neuronal cells. This is surprising because transcription of PRV LLT in vivo has been attributed to LAP1 and the initiation site was mapped downstream of the LAP1 TATA box and upstream of the LAP2 TATA box. Although LAP1 was not active in these experiments, there was a 3- to 10-fold enhancement of activity when LAP1 and LAP2 were placed in tandem. PMID- 10470262 TI - The adenovirus that causes hemorrhagic disease of black-tailed deer is closely related to bovine adenovirus-3. AB - DNA sequence data was obtained from an adenovirus previously shown to be the cause of a distinctive, fatal hemorrhagic disease of black-tailed deer in California. A 256 base fragment of the viral hexon gene was amplified by PCR from purified adenovirus preparations. The amplicon then was cloned and sequenced. Phylogenetic relationships with other mammalian adenoviruses were also determined. Although sequence analysis of this portion of the hexon gene indicates that the black-tailed deer adenovirus is closely related to bovine adenovirus-3, the biologic properties of the two viruses are clearly distinct. PMID- 10470263 TI - Analysis of the thymidine kinase genes of macropodid herpesviruses 1 and 2. AB - The nucleotide sequences of the entire protein coding regions of the thymidine kinase (TK) genes of macropodid herpesvirus type 1 (MaHV-1) and type 2 (MaHV-2) were determined. The coding region of the MaHV-1 TK gene was 984 bp long and was predicted to encode a polypeptide of 327 amino acids. The coding region of the MaHV-2 TK gene was 1020 bp long and encoded a polypeptide of 340 amino acids. Comparisons of their deduced amino acid sequences with those of fifteen other herpesviruses revealed close homology to those of other alphaherpesviruses, particularly to human herpesvirus type 1 (HHV-1) and type 2 (HHV-2). PMID- 10470264 TI - Structure of the immediate early gene of canine herpesvirus. AB - The nucleotide sequence of the immediate early (IE) gene of canine herpesvirus was determined. This gene was located in the inverted repeat regions, encoding a polypeptide of 1,383 amino acids. The predicted amino acid sequence was most closely related to that of the feline herpesvirus 1 IE protein among those of other alphaherpesviruses. DNA binding and transcriptional activation domains were found in the IE protein. A spliced region of the IE gene transcript was determined in its 5' non-coding region. PMID- 10470265 TI - Virus taxonomy--1999. The universal system of virus taxonomy, updated to include the new proposals ratified by the International Committee on Taxonomy of Viruses during 1998. PMID- 10470266 TI - Beijerinck's contribution to the virus concept--an introduction. AB - The existence of viruses was first recognized when certain pathogens were found to pass through filters that otherwise stop bacteria. Pasteur made such observations in 1887 with the pathogen of rabies, but he thought that the pathogen was a very subtle microbe. In 1886 Adolf Mayer studied the mosaic disease of tobacco plants. He was unable to observe the least trace of a microbe, but still assumed that the pathogen was a bacterium. In 1892 Iwanovsky demonstrated that tobacco mosaic was caused by an agent that passed through bacteria-proof filters but he insisted till the end of his life that the tobacco mosaic virus was a small bacterium. Similar observations were made by Loeffler and Frosch in 1898 on foot-and-mouth disease of cattle. Beijcrinck confirmed the filterability of tobacco mosaic virus but confirmed its properties in more detail and then, in 1898, firmly concluded that tobacco mosaic virus is not a microbe but a contagium vivum fluidum. His idea that a pathogen can be a soluble molecule that proliferates when it is part of the protoplasm of a living cell was revolutionary and new. This new concept has laid the foundation of virus research and directed further studies on the nature of viruses. PMID- 10470267 TI - The Prussian state and microbiological research--Friedrich Loeffler and his approach to the "invisible" virus. AB - When Loeffler took his first steps in the newly-emerging field of virology, the aim and the methods of his research activities were influenced by two different issues: 1) Loeffler was rooted in the scientific paradigm of bacteriology, but during the progress of his research on foot-and-mouth-disease (FMD) he recognized that the classical techniques derived from bacteriology were useless in identifying the agent of this disease. Thus he focussed on the properties of the pathogen and--though he could not find a method in order to visualize the 'virus' -he tried to develop a vaccine against the disease. 2) The Prussian Government was highly interested in effectively combatting FMD. In 1897 Loeffler was appointed by the Ministry of Cultural Affairs to the newly-established commission for exploring that disease. The agricultural lobbies and the public pursued the activities of the commission with a mixture of hope and serious scepticism and demanded convincing results. These circumstances caused a considerable degree of political pressure on Loeffler, pressure which determined that his research activities would take a pragmatic approach, that he would avoid sophisticated reflections and trials on the nature of the 'virus', and that his research strategies would have as a goal the development of an effective immunization. PMID- 10470268 TI - The legacy of Friedrich Loeffler--the Institute on the Isle of Riems. AB - When starting the experiments on foot and mouth disease on the Isle of Riems in October 1910, Friedrich Loeffler could continue investigations that had been interrupted in 1907 by ministerial order. Loeffler's appointment to Berlin in 1913 and his sudden death in 1915 lead to the temporary cessation of work on the Riems. With high personal creativity and many years of seminal influence, Otto Waldmann carried Loeffler's ideas further, in the selection of themes and research strategy, making the Riems a world famous place of research. Some essential elements have determined life and research on the island for decades: the development of measures against epidemics, the conception of their application, the extension of research to new diseases of economic significance, the close contacts with the veterinary practice at all times, the presentation of results to experts and the stimulating discussions in the laboratory. I will try to briefly draw a bow covering the decades of different social conditions to the present and to suggest that Loeffler's ideas, which have been improved with the years, can affect many a current decision, even though differentiated individually. PMID- 10470269 TI - The post-Loeffler-Frosch era: contribution of German virologists. AB - This presentation dealt with the contributions of German virologists in the rapid development of virology following the Loeffler-Frosch era. Thereby, only research was included which was undertaken within German institutions, even though guest scientists from other countries or international cooperative efforts have in some cases contributed to the work. Contributions to the field of veterinary virology were not considered here, since this topic was treated separately during this centennial symposium. The overview includes contributions of the very early period when interest was focussed mainly on the determination of the physicochemical properties of the fast growing number of newly detected viruses, and of the pioneering period when fundamental discoveries of the nature of viruses were made. The concepts that derived from those studies made the development of modern virology possible. Some highlights of the present period were presented describing the findings of selected virus families. This part was followed by a description of the results which were relevant to problems of how viruses become pathogens, and the role of the immune response to virus infections. Finally, attention was drawn to the contributions of molecular studies which became important not only for the field of virology but also for life sciences in general. PMID- 10470270 TI - Importance and impact of veterinary virology in Germany. AB - The causative agent of tobacco mosaic and of foot and mouth disease (FMD) were recognized in 1898 as "filterable" or "invisible"--and eventually termed "virus". Four years later the viral aetiology of yellow fever was established, and the new discipline took off. Thus animal virology started with a veterinary problem, and Germany's contribution during the following decades came mainly from the chairs of veterinary teaching and research establishments in Giessen, Munich and Hanover, the Riems Institute, and the Federal Research Institute for Animal Virus Diseases in Tubingen. From a superficial bibliometric analysis, a wide divergence in impact figures is noted, with excellent contributions in international virology journals and lesser papers in German veterinary journals. The publications in the observed time frame reveal a fascination by virion structure, physical characteristics and structure-function relationships with little work published in journals dedicated to immunology and pathogenesis. PMID- 10470271 TI - The evolution of viruses, the emergence of viral diseases: a synthesis that Martinus Beijerinck might enjoy. AB - The relentless production of viral variants and their selection for improved "fit" are seen from the perspective of the infectious disease sciences as ever changing viral phenotypes and emerging disease risks. In the Darwinian cause:effect equation, we can characterize very well the effects of mutation and selection--these are catalogued as new viral phenotypes or pathotypes. However, the selective forces themselves driving such changes remain rather mysterious. Many selective forces must be at work, acting on the virus, the host, the host population and the environment. In some instances the virus seems to test new unoccupied niches in the absence of any apparent environmental change, but usually it is clear that changes are driven by human activity. Most important must be the ever increasing density of human, domestic animal and crop plant populations and the consequent increased opportunities for transmission of viral variants. Also important must be the great changes affecting all ecosystems- these especially favor the emergence of new zoonotic viruses and viral "species jumpers." The great increase in human travel and transport carries exotic viruses, vectors and hosts around the world, again favoring viral occupation of new niches. The rise of bioterrorism adds yet another threat. Increasing numbers of emerging viral disease episodes seem to be linked to a decline in global resources for proven public health programs, agricultural extension programs, and the like, programs that have stood in the way of the spread and evolution of viral pathogens. If the relationship between viral evolution and the emergence of new viral diseases is rooted firstly in the host and the host population, then more research and resources must be directed to intervention at these levels rather than at the level of the viruses themselves. PMID- 10470272 TI - Life beyond eradication: veterinary viruses in basic science. AB - To some, the focus of research in virology entails the search for solutions of practical problems. By definition then, attention is limited to those viruses that cause disease or to exploitation of some aspect of virology to a practical end (e.g., antiviral drugs or vaccines). Once a disease is cured, or the agent eradicated, it is time to move on to something else. To others, virology offers the opportunity to study fundamental problems in biology. Work on these problems may offer no obvious practical justification; it is an affliction of the terminally curious, perhaps with the outside hope that something "useful" will come of it. To do this so-called "basic science", one must find the most tractable system to solve the problem, not the system that has "relevance" to disease. I have found that veterinary viruses offer a variety of opportunities to study relevant problems at the fundamental level. To illustrate this point, I describe some recent experiments in my laboratory using pseudorabies virus (PRV), a swine herpesvirus. PMID- 10470273 TI - Immune modulation by proteins secreted from cells infected by vaccinia virus. AB - Vaccinia virus comprises the live vaccine that was used for vaccination against smallpox. Following the eradication of smallpox, vaccinia virus was developed as an expression vector that is now used widely in biological research and vaccine development. In recent years vaccinia virus and other poxviruses have been found to express a collection of proteins that block parts of the host response to infection. Some of these proteins are secreted from the infected cell where they bind and neutralise host cytokines, chemokines and interferons (IFN). In this paper three such proteins that bind interleukin (IL)-1 beta, type I IFNs and CC chemokines are described. The study of these immunomodulatory molecules is enhancing our understanding of virus pathogenesis, yielding fundamental information about the immune system, and providing new molecules that have potential application for the treatment of immunological disorders or infectious diseases. PMID- 10470274 TI - Learning from our foes: a novel vaccine concept for influenza virus. AB - Concerted efforts to study the molecular biology of influenza viruses and the ability to genetically engineer them have dramatically advanced our understanding of the functions of influenza viral genes and gene products. The only nonstructural protein (NS1) coded for by the influenza virus was shown to possess interferon antagonist activity and thus to play an important role in countering the interferon (antiviral) response of the host following infection. Influenza A and B virus mutants with "weak" anti-interferon activity are highly attenuated because the host is able to mount an effective interferon response. It is suggested that these NS1-modified attenuated influenza viruses can induce a protective immune response and that they are ideal live virus vaccine candidates against influenza. PMID- 10470275 TI - Pathogenic aspects of measles virus infections. AB - Measles virus (MV) infections normally cause an acute self limiting disease which is resumed by a virus-specific immune response and leads to the establishment of a lifelong immunity. Complications associated with acute measles can, on rare occasions, involve the central nervous system (CNS). These are postinfectious measles encephalitis which develops soon after infection, and, months to years after the acute disease, measles inclusion body encephalitis (MIBE) and subacute sclerosing panencephalitis (SSPE) which are based on a persistent MV infection of brain cells. Before the advent of HIV, SSPE was the best studied slow viral infection of the CNS, and particular restrictions of MV gene expression as well as MV interactions with neural cells have revealed important insights into the pathogenesis of persistent viral CNS infections. MV CNS complication do, however, not large contribute to the high rate of mortality seen in association with acute measles worldwide. The latter is due to a virus-induced suppression of immune functions which favors the establishment of opportunistic infections. Mechanisms underlying MV-mediated immunosuppression are not well understood. Recent studies have indicated that MV-induced disruption of immune functions may be multifactorial including the interference with cytokine synthesis, the induction of soluble inhibitory factors or apoptosis and negative signalling to T cells by the viral glycoproteins expressed on the surface of infected cells, particularly dendritic cells. PMID- 10470276 TI - The glycoproteins of Marburg and Ebola virus and their potential roles in pathogenesis. AB - Filoviruses cause systemic infections that can lead to severe hemorrhagic fever in human and non-human primates. The primary target of the virus appears to be the mononuclear phagocytic system. As the virus spreads through the organism, the spectrum of target cells increases to include endothelial cells, fibroblasts, hepatocytes, and many other cells. There is evidence that the filovirus glycoprotein plays an important role in cell tropism, spread of infection, and pathogenicity. Biosynthesis of the glycoprotein forming the spikes on the virion surface involves cleavage by the host cell protease furin into two disulfide linked subunits GP1 and GP2. GP1 is also shed in soluble form from infected cells. Different strains of Ebola virus show variations in the cleavability of the glycoprotein, that may account for differences in pathogenicity, as has been observed with influenza viruses and paramyxoviruses. Expression of the spike glycoprotein of Ebola virus, but not of Marburg virus, requires transcriptional editing. Unedited GP mRNA yields the nonstructural glycoprotein sGP, which is secreted extensively from infected cells. Whether the soluble glycoproteins GP1 and sGP interfere with the humoral immune response and other defense mechanisms remains to be determined. PMID- 10470277 TI - Retroviruses: ancient and modern. AB - Retroviruses are transmitted in two distinct ways: as infectious virions and as 'endogenous' proviral DNA integrated in the germ line of their hosts. Modern infectious viruses such as HIV recently infected mankind from simian hosts, whereas human endogenous retroviral genomes have been present throughout old world primate evolution. Recently we have characterised novel retroviruses in humans and pigs. Human retrovirus 5 (HRV-5) is detected as an exogenous genome in association with arthritis and systemic lupus erythematosus. Porcine endogenous retroviruses (PERV) are carried in swine DNA but can be activated to produce virions that are infectious for human cells, which has implications for xenotransplantation. A brief account of HRV-5 and PERV is given here. PMID- 10470278 TI - Foot-and-mouth disease and beyond: vaccine design, past, present and future. AB - The first experimental vaccines against foot-and-mouth disease were made in 1925 by Vallee, Carre and Rinjard using formaldehyde inactivation of tongue tissue from cattle infected with the virus. This method was essentially unaltered until the late 1940s when the important experiments by Frenkel in Holland showed that the quantities of virus required for vaccine production could be obtained from fragments of tongue epithelium incubated in vitro following infection with the virus. This major step made possible the comprehensive vaccination programmes which followed in Western Europe and which, in turn, resulted in the elimination of the disease from that part of the world by 1989. This spectacular success has led many to question whether other kinds of vaccine are required to control the disease worldwide. Such reservations ignore the danger to the environment associated with the growth of large amounts of virus. This can never be a zero risk situation. Consequently, a vaccine which is not based on infectious virus as starting material has many attractions from safety considerations alone. In addition, a vaccine based on more fundamental considerations would not only be more aesthetically satisfying but could possibly provide an understanding at the molecular level of antigenic variation, still a problem in the control of the disease. The advances in our knowledge of the structure of the virus and the fragments which elicit a protective immune response now allow us to envisage a vaccine which does not require infectious virus and which protects against the multiple serotypes of the agent. Since antigenic variation is still a major problem in the control of the disease by vaccination, such a product would have important advantages over the current vaccines. PMID- 10470279 TI - Viruses and gene silencing in plants. AB - Genetic engineering of virus resistance in plants may be conferred by transgenes based on sequences from the viral genome. In many instances the underlying mechanism involves the transgenically expressed proteins. However there are other examples in which the mechanism is based on RNA. It appears that this mechanism is related to post transcriptional gene silencing in transgenic plants. This gene silencing is likely to involve antisense RNA produced by the action of a host encoded RNA dependent RNA polymerase. The natural role of this mechanism is as a genetic immune system conferring protection against viruses. There may also be a genomic role of the process reflected in RNA directed methylation of transgenes. Further understanding of this mechanism has obvious implications for virus resistance in plants. In addition the gene silencing can be used as a component of a new technology with application in functional genomics. PMID- 10470280 TI - Viroids and the nature of viroid diseases. AB - In its methodology, the unexpected discovery of the viroid in 1971 resembles that of the virus by Beijerinck some 70 years earlier. In either case, a novel type of plant pathogen was recognized by its ability to penetrate through a medium with pores small enough to exclude even the smallest previously known pathogen: bacteria as compared with the tobacco mosaic agent; viruses as compared with the potato spindle tuber agent. Interestingly, one of the two methods used by Beijerinck, diffusion of the tobacco mosaic agent into agar gels, is conceptually similar to one method used to establish the size of the potato spindle tuber agent, namely polyacrylamide gel electrophoresis. Further work demonstrated that neither agent is an unusually small conventional pathogen (a microbe in the case of the tobacco mosaic agent; a virus in the case of the potato spindle tuber agent), but that either agent represents the prototype of a fundamentally distinct class of pathogen, the viruses and the viroids, respectively. With the viroids, this distinction became evident once their unique molecular structure, lack of mRNA activity, and autonomous replication had become elucidated. Functionally, viroids rely to a far greater extent than viruses on their host's biosynthetic systems: Whereas translation of viral genetic information is essential for virus replication, viroids are totally dependent on their hosts' transcriptional system and, in contrast to viruses, no viroid-coded proteins are involved. Because of the viroids' simplicity and extremely small size they approach more closely even than viruses Beijerinck's concept of a contagium vivum fluidum. PMID- 10470281 TI - Improvement of survival over time for colon cancer patients by anatomical sub sites. AB - The aims of this study are to describe and to evaluate improvement of survival over time for colon cancer patients by anatomical sub-sites. Data on 661 patients newly diagnosed as having colon cancer at Osaka Medical Center for Cancer and Cardiovascular Diseases from 1978 to 1991 were examined in this study. Corrected survival was calculated with the Kaplan-Meier method according to the period of diagnosis: early period (1978-84) and later period (1985-91). Factors concerning the difference in survival between the two periods were examined with the Cox proportional hazards regression model according to sub-site. Five-year corrected survival of the patients with left colon cancer improved significantly (60 to 72%; P < 0.01), probably due to advances in treatment, while that of patients with transverse colon cancer also improved significantly (39 to 67%; P < 0.01), mainly because of progress in diagnosis. The five-year corrected survival of those with right colon cancer did not increase (57 to 46%; P = 0.14), owing to lack of improvement in stage at diagnosis. Among the three sub-sites, the right showed the worst five-year survival in the later period. We concluded that survival of patients with right colon cancer, differing from the other anatomical sub-sites, did not improve, possibly because of lack of symptoms. The screening programs for colon cancer introduced in Japan in 1992 may be expected to improve the survival of patients with colon cancer, including that of the right colon. PMID- 10470282 TI - Genotype difference of aldehyde dehydrogenase 2 gene in alcohol drinkers influences the incidence of Japanese colorectal cancer patients. AB - A case-control study was conducted to explore the possible etiologic role of alcohol and aldehyde dehydrogenase 2 (ALDH2) gene among Japanese colorectal cancer patients. Information on their drinking, smoking and dietary habits was collected from 265 colon and 164 rectum cancer patients, and 794 non-cancer patients as a control group. Genotypes of the ALDH2 gene at codon 487, glutamic acid (ALDH2*1) as a wild-type or lysine (ALDH2*2) as a mutated type with reduced enzyme activity, were analyzed by polymerase chain reaction in 160 colon and 110 rectum cancer patients and 121 control persons. Univariate analysis with the chi 2 statistical test showed that heavy alcohol drinking (P < 0.01), frequent meat intake (P < 0.001), and irregular (P < 0.01), hasty (P < 0.01) and excessive (P < 0.001) eating habits were associated with the incidence of both colon and rectum cancers, whereas heavier smoking (P < 0.05) and infrequent fish (P < 0.03) and fruit (P < 0.01) intake were solely associated with incidence of rectum cancer. Infrequent green vegetable intake was not correlated with the incidence of colorectal cancer. Multivariate unconditional logistic regression analysis confirmed the association of alcohol consumption (P < 0.01) and meat intake (P < 0.05). Homozygous and heterozygous carriers of ALDH2*2 allele tended to be found in colon (trend P = 0.04) but not in rectum cancer patients compared to controls. Risk elevation for colon cancer due to alcohol consumption was pronounced among the heterozygotes and it was statistically significant especially for distal colon cancer (trend P = 0.02). We conclude that alcohol consumption is a risk factor for colorectal cancer and the risk can be enhanced in ALDH2 heterozygotes. PMID- 10470283 TI - Development and distribution of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP)-induced aberrant crypt foci in the rat large intestine. AB - Aberrant crypt foci (ACF) are generally considered to be preneoplastic lesions for colon cancer. To assess their induction by 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP), a colon carcinogen, we performed a sequential study of ACF morphology and localization. F344 male rats were given PhIP, and methylene blue-stained colon epithelium and isolated crypts were analyzed at weeks 12, 25, 50, and 75. Each crypt was classified into 2 groups, "single" with round bottoms and "bifurcating" displaying V-shaped clefts (indicating proliferation). In combination with the number of crypts in an ACF, this classification was a good indicator for the generation of ACF in line with the fission mechanism of growth. Increasing numbers of crypts in ACF through weeks 12 to 75 and decreased percentages of ACF with bifurcating crypts at the late time points indicated that proliferation of crypts occurs predominantly during the early stages. The distribution pattern showed a significant shift (P < 0.000005) from the distal to the proximal part of the large intestine between weeks 25 and 50. Adenocarcinomas were first found to develop at week 50 in the ascending colon and cecum where bifurcating crypts were generally lacking at weeks 12 and 25. These data suggest the existence of (1) proliferating ACF which contains bifurcating crypt(s) and (2) quiescent or senescent ACF which consists of only single crypts. PMID- 10470284 TI - Preventive effects of Glycyrrhizae radix extract on estrogen-related endometrial carcinogenesis in mice. AB - Short- and long-term experiments were conducted to examine the effects of Glycyrrhizae radix (Gl radix) extract on mouse endometrial carcinogenesis. Gl radix treatment (2 weeks) decreased the levels of c-fos/jun mRNA and the corresponding oncoproteins induced by estradiol-17 beta (E2) in castrated mice uteri, as determined by reverse transcription-polymerase chain reaction and Southern blot analysis, and immunohistochemical methods, respectively. For the long-term assays, 98 female ICR mice were given N-methyl-N-nitrosourea (MNU) solution (1 mg/100 g body wt.) and normal saline (as controls) into their left and right uterine corpora, respectively. They were divided into four groups as follows: group 1 was given 0.625% Gl radix- and 5 ppm E2-containing diet; group 2, 5 ppm E2-containing diet; group 3, 0.625% Gl radix-containing diet; and group 4, the basal diet alone. Gl radix treatment significantly decreased uterine weights and the incidences of uterine endometrial atypical hyperplastic and malignant lesions. It is suggested that Gl radix has inhibitory effects on E2 related endometrial carcinogenesis in mice, through suppression of estrogen induced c-fos/jun-expressions. PMID- 10470285 TI - Mechanistic aspects of green tea as a cancer preventive: effect of components on human stomach cancer cell lines. AB - It is now well accepted that (-)-epigallocatechin gallate (EGCG) inhibits carcinogenesis in the digestive tract in rodents. To understand the mechanisms of anticarcinogenesis, we first studied growth inhibition by EGCG in human stomach cancer cell lines established at Seoul National University (SNU cell lines). Inhibition by EGCG of [3H]thymidine incorporation into eight SNU cell lines was examined, in relation to transforming growth factor-beta (TGF-beta) responsiveness. Various tea polyphenols derived from green tea and black tea induced growth inhibition and apoptosis of human stomach cancer cell line KATO III, and inhibition of tumor necrosis factor-alpha (TNF-alpha) release from the cells, in the order of (-)-epicatechin gallate (ECG), EGCG, (-)-epigallocatechin (EGC), teaflavins (TF) and (-)-epicatechin (EC). In addition, we demonstrated that EGCG inhibited TNF-alpha gene expression in KATO III cells, as well as okadaic acid-induced AP-1 and NF-kappa B activation. The inhibitory potencies of EGCG for AP-1 and NF-kappa B binding to DNA were different between KATO III cells and mouse fibroblast cell line BALB/3T3. Thus, EGCG and other tea polyphenols may interact with various transcription factors, in addition to AP-1 and NF-kappa B, in nuclei of various cells, resulting in inhibition of TNF-alpha gene expression and TNF-alpha release. PMID- 10470286 TI - Mxi1 mutations in human neurofibrosarcomas. AB - Mxi1 is thought to negatively regulate Myc function and may therefore be a potential tumor suppressor gene. Little effort has yet been made to find alterations involving this gene in human solid tumors. We screened 31 human gastric cancers, 7 esophageal cancers, 85 bone and soft tissue tumors of various types, including 4 neurofibrosarcomas. We also examined 29 human tumor cell lines consisting of 12 esophageal cancers, 7 glioma/glioblastomas and 10 others for Mxi1 mutations in exons 1, 2, 4 (HLH domain), 5 and 6. Polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and subsequent sequencing revealed three distinct polymorphisms in the intron-exon boundary upstream from exon 6. We discovered a missense mutation, GCA to GTA (Ala 54 Val), in exon 2 in a neurofibrosarcoma patient (case 1), two missense mutations, AAA to CAA (Lys 118 Gln) and GAA to GGA (Glu 154 Gly) in exon 5 of another neurofibrosarcoma patient (case 2), and 3 amino acid substitutions, GTG to GCG (Val 179 Ala), GTT to GCT (Val 181 Ala) and TTC to CTC (Phe 186 Leu), in a third neurofibrosarcoma patient (case 3). In case 3, loss of heterozygosity was also demonstrated by informative (TTC)3/(TTC)2 polymorphism. Our data demonstrate that mutations occur in the Mxi1 gene in neurofibrosarcoma. Missense mutations in the functional domain of Mxi1 in these cases may be involved in the pathogenesis of neurofibrosarcoma. PMID- 10470287 TI - Inhibitory role of plasminogen activator inhibitor-1 in invasion and proliferation of HLE hepatocellular carcinoma cells. AB - Plasminogen activator inhibitor (PAI)-1, a serine protease inhibitor, inactivates urokinase-type plasminogen activator (uPA) and regulates degradation of the extracellular matrix; whether it functions for or against tumor progression, however, has been the subject of controversy. To assess the role of PAI-1 in invasion and proliferation of hepatocellular carcinoma (HCC) cells, HLE cells were transfected with a vector capable of expressing an antisense PAI-1 transcript. Analysis of seven stably transfected clones (PAI-1-) showed reductions of 81% in PAI-1 mRNA by northern blot analysis and 63% in the cellular PAI-1 antigen level by enzyme-linked immunosorbent assay (ELISA). There was no change in the levels of secreted PAI-1 or PAI-2. The activity of cellular uPA increased by 54%, without change in the protein level or the secreted uPA activity evaluated by ELISA. Morphologically, PAI-1 antisense induced a spindle shape with narrower cytoplasmic processes in HLE cells. The forced inhibition of PAI-1 increased the invasion and the growth of PAI-1- cells by 75% and 82%, respectively. These results suggest that PAI-1 plays a role in inhibiting invasion and proliferation, and the balance between uPA and PAI-1 expression is important to assess the invasiveness of HCC cells. PMID- 10470288 TI - Detection of disseminated urothelial cancer cells in peripheral venous blood by a cytokeratin 20-specific nested reverse transcriptase-polymerase chain reaction. AB - More than half of all patients with invasive urothelial cancer subsequently develop metastatic disease even after radical resection of the primary cancer. In these patients, neoplastic cells may be disseminated prior to or during the operation. A nested reverse transcriptase-polymerase chain reaction (nested RT PCR) assay which amplifies cytokeratin (CK) 20 transcripts was used to detect cancer cells in the peripheral blood of urothelial patients. This assay was able to detect 10 bladder cancer cell line cells in a sample of ten million peripheral blood mononuclear (PBMN) cells. CK 20-specific signals were detected in 9 (22.5%) of 40 PBMN cell samples prepared from 40 urothelial cancer patients in relation to the tumor stage, including 0/13 patients with a superficial tumor, 4/21 (19%) with a regionally invasive tumor and 5/6 (83%) with a metastatic tumor (P = 0.0002 in chi 2 test). No signals were detected in any of 25 healthy donor PBMN cell samples. The present results indicate that the CK 20 RT-PCR assay is applicable for detection of urothelial cancer cells in the peripheral blood. The assay also confirms that hematogenic dissemination occurs in invasive urothelial cancers but rarely in superficial ones. PMID- 10470289 TI - Indomethacin suppresses the growth of colon 26, Meth-A and FM3A tumors in mice by reducing the prostaglandin E2 content and telomerase activity in tumor tissues. AB - The antitumor effect of indomethacin on Colon 26, Meth-A and FM3A tumors was investigated in mice. The prostaglandin E2 content in tumor tissues was assayed to find out if indomethacin acts on tumors, and the telomerase activity in tumors and somatic tissues (testis, liver, spleen and colon) was also monitored during indomethacin treatment. Growth of Colon 26, Meth-A and FM3A tumors was significantly (P < 0.001-0.05) suppressed by indomethacin compared to the untreated controls. The prostaglandin E2 content in the three tumors was markedly (P < 0.001) reduced by indomethacin. Telomerase activity in Colon 26 and FM3A tumors was significantly (P < 0.001) lower than that of untreated tumors (80% and 45% decrease versus the controls, respectively), and the activity in Meth-A tumor was slightly decreased (10% decrease versus the control) by indomethacin. Telomerase activity in the somatic tissues was not significantly affected by indomethacin. In summary, this study shows the effectiveness of indomethacin as an antitumor agent against three types of tumors, and suggests that indomethacin affects telomerase activity in tumors in vivo. PMID- 10470290 TI - Prevention by chitosan of myelotoxicity, gastrointestinal toxicity and immunocompetent organic toxicity induced by 5-fluorouracil without loss of antitumor activity in mice. AB - We examined the antitumor activity and side effects (myelotoxicity, immunocompetent organic toxicity and gastrointestinal toxicity) of combined treatment with the cancer chemotherapy drug 5-fluorouracil (5-FU) and dietary fiber chitosan in sarcoma 180-bearing mice. 5-FU (12.5 mg/kg x 2/day) plus chitosan (150, 375 and 750 mg/kg x 2/day) inhibited the tumor growth as well as 5 FU alone. Chitosan (150 and 750 mg/kg x 2/day) blocked the reduction of blood leukocyte number caused by 5-FU administration, and it prevented the injury of the small intestinal mucosa membrane and delayed the onset of diarrhea induced by 5-FU. Furthermore, chitosan (750 mg/kg x 2/day) prevented the reduction of spleen weight induced by 5-FU in sarcoma 180-bearing mice, and the reduction of lymphocyte and CD8+ T cell numbers induced by 5-FU was also prevented by the oral administration of chitosan (750 mg/kg x 2/day) in C57BL/6 mice. Chitosan (150 and/or 750 mg/kg x 2/day) reduced the 5-FU incorporation into RNA fractions of small intestine and spleen without affecting the 5-FU incorporation into the tumor in sarcoma 180-bearing mice. These findings suggest that prevention of the 5-FU side effects by chitosan might be partly due to the selective inhibition of 5-FU uptake into the small intestine and spleen, resulting in the reduction of immune function toxicity, myelotoxicity and gastrointestinal toxicity of 5-FU. Therefore, it is concluded that the combination of chitosan and 5-FU might be useful for the prevention of side effects such as gastrointestinal toxicity, immunotoxicity and myelotoxicity caused by 5-FU. PMID- 10470291 TI - Membrane transport and antitumor activity of pirarubicin, and comparison with those of doxorubicin. AB - We have compared the membrane transport and antitumor activity of pirarubicin with those of doxorubicin in M5076 ovarian sarcoma, which exhibits low sensitivity to doxorubicin. Pirarubicin was rapidly taken up by M5076 cells and the intracellular concentration of pirarubicin reached more than 2.5-fold that of doxorubicin. In terms of the 50% cell growth-inhibitory concentration in vitro, pirarubicin was more effective than doxorubicin. Thus, the intracellular concentration influenced the cytotoxicity of these anthracycline agents. On comparison of the nuclear uptake of pirarubicin and doxorubicin, the nucleus/cell ratio of pirarubicin was found to be about 40%, whereas that of doxorubicin reached more than 80%. As the intranuclear concentration of pirarubicin is dependent on nuclear transport, the increases in not only cell membrane transport, but also nuclear membrane transport contributed to the enhancement of the efficacy of pirarubicin. In M5076 solid tumor-bearing mice, pirarubicin reduced the tumor weight to 60% of the control level, although doxorubicin had no effect. These results were supported by the intracellular uptake of pirarubicin. Moreover, theanine, which inhibited the pirarubicin efflux from M5076 cells, increased by 1.3-fold the pirarubicin concentration in the tumor and enhanced the therapeutic efficacy of pirarubicin 1.7-fold. In conclusion, our results suggest that an increase in the concentration of an anthracycline derivative in tumor cells due to alteration of cell membrane transport results in enhancement of the antitumor activity. PMID- 10470292 TI - Contribution of the nucleoside transport system to doxorubicin transport in HL60 cells but not in mononuclear cells. AB - Previously, we reported that pirarubicin (THP), an anthracycline, was transported, at least in part, via a nucleoside transport system in human leukemic HL60 cells, but not in mononuclear cells (MNCs). In this study, the contribution of the nucleoside transport system to the transport of other anthracyclines, doxorubicin (DOX), daunorubicin (DNR) and idarubicin (IDA), in HL60 cells and MNCs was investigated. The experiments were performed after both types of cells had been pretreated with a metabolic inhibitor, 2,4-dinitrophenol, to deplete cellular ATP. The DOX uptake by HL60 cells was partially inhibited by inhibitors of equilibrative nucleoside transporters. In HL60 cells, moreover, the uptake of DOX depended on an inwardly directed Na(+)-gradient, and was inhibited by concentrative nucleoside transporters, but there was no change in the DNR or IDA uptake under any of these conditions. On the other hand, the uptake of the three drugs by MNCs was not affected by any inhibitors of the nucleoside transporters, and there was no dependence of the uptake on an Na(+)-gradient. These results suggested that DOX, but not DNR or IDA, was partially transported in HL60 cells via the nucleoside transport system, whereas in MNCs the system did not contribute to the uptake of any of these three drugs. Thus, nucleoside transport systems contributing to the transport of anthracyclines may be different among different derivatives and cell types. PMID- 10470293 TI - Multidrug resistance phenotype in the RMS-GR human rhabdomyosarcoma cell line obtained after polychemotherapy. AB - Classical cytotoxic treatment of rhabdomyosarcoma (RMS), the most common soft tissue malignacy in children, is often accompanied by significant morbidity and poor response. Chemotherapy may induce multidrug resistance (MDR) associated with the expression of P-glycoprotein, a drug efflux pump which modifies the sensitivity of tumoral cells to drugs. To analyze MDR in RMS we used the RMS-GR cell line, obtained from an embryonal rhabdomyosarcoma treated in vivo with polychemotherapy. The RMS-GR cells showed cross-resistance to vincristine, doxorubicin and actinomycin D, the drugs of choice in the conventional treatment of RMS. Polymerase chain reaction (PCR) analysis showed that these RMS cells overexpressed mdr1/P-glycoprotein. The pattern of resistance and the level of P glycoprotein expression were similar to those found in the resistant RMS TE.32.7.DAC cell line obtained in vitro. Southern blot analysis showed that mdr1 overexpression was not due to amplification of the gene. Our results showed that the in vivo treatment of embryonal RMS may induce an MDR phenotype mediated by mdr1/P-glycoprotein in RMS cells. PMID- 10470294 TI - Antitumor effect on human gastric cancer and induction of apoptosis by vascular endothelial growth factor neutralizing antibody. AB - Induction of apoptosis by antiangiogenic therapy has been suggested as a new anticancer strategy. To clarify the mechanism of the antitumor effect achieved by inhibition of vascular endothelial growth factor (VEGF), which is a major mediator of angiogenesis, we used an orthotopic transplantation model of human gastric carcinoma line (MT2) treated with a monoclonal VEGF neutralizing antibody (VEGF Ab). We histologically examined the microvessel density (MVD) and the apoptotic index (AI) in this model. Transplanted tumor growth was significantly inhibited by the VEGF Ab (P = 0.03), and there was a significant decrease in the number of mice with liver metastasis (P = 0.004). The MVD detected by immunohistochemical staining with ER-MP12 antibody was 33.6 +/- 8.0 in the control group and 21.1 +/- 5.4 in the treated group, and the difference was significant (P < 0.0001). The AI values of the control and treated groups were 4.73 +/- 1.11 and 7.26 +/- 1.62, respectively, and this difference is also significant (P < 0.0001). However, the expression of VEGF mRNA in transplanted tumors did not show a significant difference between the control and treated groups. These results suggest that the antitumor effect of the VEGF Ab on human gastric carcinoma is exerted by inducing mild hypoxia followed by apoptosis, which does not influence VEGF mRNA expression in the carcinoma. PMID- 10470296 TI - South Africa will not meet WHO TB targets. PMID- 10470295 TI - Use of antiviral agents in pregnant women with HIV infection. PMID- 10470297 TI - No to psychologists' right to prescribe. PMID- 10470298 TI - Stenting--protocols needed! PMID- 10470299 TI - Affordable psychiatric medication. PMID- 10470300 TI - Making a difference. PMID- 10470301 TI - Viagra soft on sales. PMID- 10470304 TI - Haemophilus influenzae type B vaccine. PMID- 10470305 TI - Antimicrobial therapy for serious bacterial infections in children in a primary care setting--a poll of experts. PMID- 10470306 TI - Rheumatoid factor--does the patient have arthritis? PMID- 10470307 TI - Gastro-intestinal scleroderma. PMID- 10470308 TI - A mood of mild self-congratulation. PMID- 10470309 TI - Emerging and re-emerging diseases--epidemic enterohaemorrhagic infections 100 years after Shiga. PMID- 10470310 TI - Beyond the haze of the tobacco bill debate. PMID- 10470311 TI - Infant mortality in the rural Hlabisa health district--estimation using the previous birth technique. PMID- 10470312 TI - D8/17 in obsessive-compulsive disorder and trichotillomania. PMID- 10470313 TI - An essential hospital package for South Africa--selection criteria, costs and affordability. AB - BACKGROUND: In 1995 the Committee of Enquiry into National Health Insurance (NHI) recommended that formally employed individuals and their employers be required to fund at least a minimum package of hospital cover for workers and their dependents. This has recently been echoed in a Department of Health policy paper on social health insurance. This research aims to define and cost a minimum package of essential hospital care for competing (public and private) health insurers in South Africa. CRITERIA FOR PACKAGE DEFINITION: Based on the objectives implict in the NHI Committee report, the following criteria were used to define the essential package: (i) the extent to which there was another appropriate responsible party who should pay for treatment; (ii) the degree of discretion in deciding whether or not to provide treatment (roughly equivalent to 'urgency'); and (iii) the cost and effectiveness of treatment. RESULTS: On the basis of the above criteria, 396 out of 598 possible interventions were included in the package. Using local mine hospital and private sector utilisation rates and mine hospital cost data, it was estimated that the essential inpatient package would cost around R502 per enrollee per year, using 1998 prices, for a working age population and their dependents. Age-sex standardised outpatient care costs in the mine hospital population studied were estimated at R183 per person per year. It was therefore estimated that the total inpatient and outpatient hospital package would cost around R685 per person per year. CONCLUSIONS: The results presented in this paper are intended to inform the process of defining a national essential hospital benefit package. Assuming that contributions were proportionally related to income, and that costs should not exceed 6% of wages, the package should be affordable to all of those earning above R20,000 per year. Significant additional work is required, firstly at a technical level to assess the appropriateness of the prioritization approach used here, and secondly to take the debate around essential hospital benefits to broader political and public forums. PMID- 10470314 TI - The McCord-Christian medical Fellowship vocational training programme. AB - The first 3 years' experience of a vocational training programme for doctors based at McCord Hospital in Durban is described and reviewed. The stated aim of the programme is to equip doctors to 'serve the under-served according to the example of Christ' and to produce caring, competent and committed doctors for areas of need. PMID- 10470315 TI - The procedural skills of rural hospital doctors. AB - OBJECTIVES: To describe the spectrum of procedures performed by general medical officers in South African rural hospitals, and the attitudinal issues associated with these tasks. DESIGN: A descriptive study combining quantitative and qualitative methods: a quantitative audit of operating theatre records of 15 rural hospitals in KwaZulu-Natal and 4 rural hospitals in Northern Province, and a qualitative analysis of 8 focus group discussions in which rural hospital doctors discussed their procedural work. OUTCOME MEASURES: Quantitative results included patient age, type of anaesthetic, type of procedure, and whether a separate anaesthetist was present or not for each recorded procedure performed in the operating theatre. Qualitative results comprised major and minor themes that arose from the focus group discussions. RESULTS: Quantitative results revealed a high proportion of obstetric and general surgical procedures, often performed by a single doctor acting as anaesthetist as well as surgeon. The range of surgical procedures undertaken varied widely between hospitals. Anaesthetic usage patterns revealed extensive use of ketamine and spinal anaesthetics, with relatively more general anaesthetics being administered in the Northern Province. Qualitative results revealed two scenarios in the rural hospital situation: one where doctors felt that they were coping and learning from the work under the supervision of peers or senior colleagues, and the other where they felt stressed by being alone and having to deal with emergencies, especially when short-staffed. CONCLUSIONS: The broad range of skills demanded in rural hospital practice requires specific preparation and ongoing support. Without it, a vicious cycle is established leading to poor output and morale. The phenomenon of the single-handed anaesthetist-surgeon that emerged from this study deserves specific attention, particularly with regard to staff shortages in rural areas. Under circumstances of adequate staffing and support, the rural hospital is an ideal learning site for generalists. This study has significant implications for the proposed introduction of obligatory vocational training for all medical graduates. PMID- 10470316 TI - Utilisation of predictive, prenatal and diagnostic testing for Huntington's disease in Johannesburg. AB - CONTEXT: Huntington's disease (HD) is a dominantly inherited condition in which the gene defect is known. As such individuals in at-risk families can be tested before symptoms occur, prenatally, or after symptoms appear to confirm the diagnosis. OBJECTIVES: To investigate the utilisation and sequelae of the predictive, prenatal and diagnostic services offered to families with suspected Huntington's disease. DESIGN: A retrospective design was used. The 1975-1997 records of the Department of Human Genetics for all families with a history of HD presenting for genetic counselling and DNA analysis were studied. SETTING: Department of Human Genetics, South African Institute for Medical Research and University of the Witwatersrand, Johannesburg. SUBJECTS: There were 30 at-risk (50% risk) subjects for predictive testing, 7 women (10 pregnancies) for prenatal testing, and 52 subjects for diagnostic testing. OUTCOME MEASURES: These were provided by the results from molecular studies and by the action taken by subjects after a predictive or prenatal result was given. RESULTS: Altogether 15 (50%) subjects for predictive testing had a positive result, but none had serious psychiatric sequelae. Two women were found to be carrying an affected fetus and both requested pregnancy termination. Of 52 diagnostic tests, 33 (63%) were positive. CONCLUSION: The service was used appropriately, and there were no traumatic incidents following positive results. There was no genotypic or sex bias in subjects presenting for testing. Black and white patients were equally likely to be positive for HD on diagnostic testing. The families appreciated the service and found it useful in the detection and prevention of HD. PMID- 10470318 TI - The cost-effectiveness of isotretinoin in the treatment of acne. Part 2. A chronic medication plan profiling study. AB - OBJECTIVES: Aspects of current clinical practice, needed for a pharmaco-economic model of isotretinoin, were determined from an acne-profiling study on chronic medication plan data. The patient sample was analysed as a whole and as two subgroups, representing isotretinoin and oral antibiotic patients. The study focused on the prevalence of the condition, patient age and gender distributions, and pharmacotherapeutic patterns and costs. METHODOLOGY: Anonymous patient records from the Pharmaceutical Benefit Management (Pty) Ltd, Cape Town, chronic medication plan were screened and analysed using descriptive and inferential statistical methods. For all hypothesis tests, the significance level (alpha) was set a priori at 5% (0.05). In order to determine the distribution of costs among patient groups, cost-concentration curves were constructed. RESULTS: A total of 5,198 patient records were analysed, representing 3,546 acne patients (2.7% of applicants to the plan). The majority of patients were female (72.2%, median age 23 years), with male patients treated at a younger age (median age 19 years). The cyproterone/oestrogen combination was found to be the most frequently prescribed agent (25.2% of script items), followed by isotretinoin (15.6%). The oral antibiotics as a group accounted for 24.9% of script items, the leading agents being minocycline (10.1%) and doxycycline (7.1%). Isotretinoin (44.1%) represented the greatest expense, followed by hormonal therapies (24.7%) and oral antibiotics (16.7%). Female isotretinoin patients (55.1%) were older (median age 25 years), while the male patients were of an equivalent age to the collective sample. Patients treated with oral antibiotics represented the youngest group of patients (median age 20 years), with 57.8% being female. The annual pharmacotherapy expense per patient on oral antibiotics was R1 971.81 compared with R2 670.20 for all patients, and R6 140.81 for the isotretinoin group. For each of these groups there was an uneven cost distribution among patients. CONCLUSION: The treatment of acne represents a significant burden to funders, estimated at R7.2 million per 100,000 beneficiaries. The distinct age and gender distribution patterns, together with the uneven distribution of costs among patients, provide valuable information for the treatment and management of the condition. This information indicates that the introduction of clinically based, focused management principles supported by sound pharmaco-economic arguments will be required to manage acne effectively. PMID- 10470317 TI - The cost-effectiveness of isotretinoin in the treatment of acne. Part 1. A meta analysis of effectiveness literature. AB - OBJECTIVES: This paper reports the results of a meta-analysis of isotretinoin treatment in moderate to severe acne. It forms part of a comprehensive investigation into the cost-effective treatment of acne in South Africa and as such establishes the clinical foundation for an economic model of acne management. This foundation includes an evaluation of the daily dosages, treatment durations, success rates, clinical effectiveness and relapse rates reported in published trials since 1981. METHODOLOGY: A predetermined protocol for the study established the scope, appropriate inclusion and exclusion criteria for peer-reviewed data, and the statistical rigour that would be applied to the selected data. Following an extensive literature search, data reflecting the effectiveness of isotretinoin were extracted, statistically assessed, described and reported. The combinability of the data was confirmed using analyses of variance and chi-square tests, as applicable. RESULTS: Isotretinoin consistently proved to be a highly effective agent in the treatment of moderate to severe acne vulgaris. The response rate determined by the meta-analysis indicated a clinical cure in 84.22% to 86.71% of patients treated. From the data considered, the average treatment duration was calculated to be 17.9 weeks (4 months). The relapse rate was low (21.45%) and dose-dependent. Optimal results were achieved by treating patients with a daily dose of 1 mg/kg and treating to a target cumulative dose of 120 mg/kg over the treatment duration. CONCLUSION: The results of this meta-analysis support the continued use of isotretinoin in the treatment of acne. The results are important in the field of pharmaceutical benefit management where they will assist in the optimal management of this health condition. The results will be used to develop a pharmaco-economic model to evaluate the various treatment regimens used for acne in South Africa. PMID- 10470319 TI - The cost-effectiveness of isotretinoin in the treatment of acne. Part 3. A cost minimisation pharmaco-economic model. AB - OBJECTIVES: The cost-effectiveness of systemic isotretinoin therapy in the treatment of moderate to severe acne was assessed in a comparative cost minimisation analysis. Systemic isotretinoin at the recommended daily dose of 1 mg/kg (cumulative dose of 120 mg/kg) was compared with: (i) oral antibiotics taken as chronic medication; (ii) a combination of chronic oral antibiotics and anti-androgen therapy; and (iii) isotretinoin prescribed only after two failed courses of oral antibiotics, as per South African guidelines. The perspective taken was that of the funder of health care, and the resources used were funder charges as a proxy for costs. METHODOLOGY: Statistical and epidemiological data as well as relevant costs from the previously reported meta-analysis and profiling study for acne therapy were used as the clinical basis for the construction of a cost-minimisation model. Additional costs were sourced from published pharmaceutical retail prices and professional rates. The South African treatment guidelines were used to define the frequencies associated with physician visits and pathology testing. Standard statistical methods were applied, as appropriate. From the above, a modified Markov process was used to model the costs associated with the four comparator treatment regimens over a period of 120 months. Suitable clinical and economic endpoints were defined so that comparison could be made between regimens. RESULTS: Assuming a relapse rate of 21.45%, it was found that isotretinoin therapy compares favourably with the other regimens. After 50 and 35 months, systemic isotretinoin cumulative costs were less than those incurred in oral antibiotic and oral antibiotic/anti androgen therapy, respectively. For the stepped therapy of oral antibiotics followed by systemic isotretinoin, these break-even periods were 56 and 39 months, respectively. The cost per successfully treated patient receiving isotretinoin was R8941. This compares well with the cost for those patients receiving chronic oral antibiotics, which after 5 years amounted to R10 428 per patient. Sensitivity analyses proved these findings to be robust to variations in the isotretinoin relapse rate, and the cost of oral antibiotic therapy and the concomitant use of topical therapies. CONCLUSION: From the cost-minimisation model it is clear that where systemic isotretinoin is clinically indicated, the sooner such therapy is initiated the more cost-effective the outcome will be. If isotretinoin is prescribed on diagnosis of moderate to severe acne, then the cost of treatment is significantly reduced in the long term when compared with standard chronic oral antibiotic therapy. PMID- 10470320 TI - Etiopathogenesis of acute pancreatitis. AB - Acute pancreatitis is a disease that has many causes. Each cause seems to affect the acinar cell in some way that results in the premature activation and retention of potent proteolytic enzymes. These activated enzymes then injure the acinar cell and cause the immediate release of cytokines and activate the complement system. Together, these molecules attract and sequester inflammatory cells, in particular neutrophils, which causes further secretion of cytokines, free radicals, and other vasoactive molecules, such as nitric oxide. We propose that the released inflammatory molecules induce local effects, such as pancreatic edema and necrosis, and systemic complications, such as hypotension, tachycardia, fever, capillary leak syndrome, and hypoxia. The cytokines released in the pancreas also stimulate apoptosis, further enhancing the cell death response in pancreatitis. Much of the current research is aimed at understanding the links between these series of events and finding agents that can modulate the cascade of events involved in pancreatitis. What is promising in this endeavor is that the response produced with pancreatitis is nearly identical with all etiologies, suggesting that therapy may not have to be specific to a particular cause. The mechanistic models of AP presented herein are supported by preliminary clinical studies that suggest that protease and cytokine inhibitors may improve the course of AP in specific clinical settings. PMID- 10470321 TI - Hereditary pancreatitis. Gene defects and their implications. AB - Hereditary pancreatitis is a rare condition characterized by acute and chronic pancreatitis transmitted in an autosomal dominant fashion. There also is an epidemiologic link to pancreatic cancer in some affected families. Failure of a secondary brake mechanism responsible for inactivation of prematurely activated cationic trypsin in acinar cells seems to be the fundamental defect in type I hereditary pancreatitis (R117H cationic trypsin), and also may explain the pathogenesis of type II hereditary pancreatitis (N211 cationic trypsin). The diagnosis is made based on clinical history and, in certain cases, by molecular diagnostic testing for these gene defects. Medical management of acute and chronic hereditary pancreatitis currently does not differ from that of nonhereditary AP. As in nonhereditary pancreatitis, the surgical approach must be tailored to the individual problem, with an understanding that disease restricted to the head of the gland is atypical and that residual acinar tissue continues to drive the disease state. Although diagnosis and management of pancreatic adenocarcinoma are similar in this cohort, the increased age-accumulated risk suggests that thoughtful screening protocols eventually may be clinically and cost-effective. PMID- 10470322 TI - Are mutations in the cystic fibrosis gene important in chronic pancreatitis? AB - The leading causes of chronic pancreatitis are alcohol and idiopathic pancreatitis. The importance of genetic factors in chronic pancreatitis has been uncertain. Recently, however, it was learned that many patients with idiopathic chronic pancreatitis have mutations of the cystic fibrosis gene. This article reviews the evidence that links mutations of this gene to unexplained pancreatitis, and discusses the implications of this for the evaluation, pathogenesis, classification, and possible prevention of pancreatitis. PMID- 10470323 TI - Staging of acute pancreatitis. Is it important? AB - Staging of acute pancreatitis is important for selection of patients for clinical trials, comparison of results between centers, and the early identification of patients who may benefit from therapeutic intervention or transfer to a specialist unit. The APACHE-II score and other multiple-factor scoring systems are widely used for the first two indications, and of these, the APACHE-II score provides the best accuracy at an early stage in the course of the illness. Presently, however, no system provides sufficient predictive power to facilitate clinical decision making. At a time of increasing pressure to involve specialist units at an early stage in the management of these complex patients, a pressing need to identify a system for accurate early staging of acute pancreatitis remains. PMID- 10470324 TI - Interventional management of peripancreatic fluid collections. AB - This article discusses the interventional management of peripancreatic fluid collections, focusing on endotherapy and surgical management. Anatomy and principles to decide management also are reviewed. PMID- 10470325 TI - Nonsurgical management of acute pancreatitis. Use of antibiotics. AB - In severe AP, infected necrosis is the leading cause of death. Prevention of pancreatic infection is the major goal in the treatment of patients with necrotizing pancreatitis. Adequate early antibiotic therapy seems to be promising in these patients. Their role and the optimal timing of the antibiotic therapy (e.g., benefit of prophylactic application) are discussed. Preliminary results of a study in patients with infected pancreatic necrosis and exclusively or primarily conservative treatment also are presented. PMID- 10470326 TI - The potential role of therapeutic cytokine manipulation in acute pancreatitis. AB - The central, detrimental role of the inflammatory cytokines IL-1 and TNF and the biologically active phospholipid PAF in the pathogenesis of AP has been established over the past 8 years. A number of antagonists to these mediators have been used successfully in the laboratory setting and are currently being examined in prospective randomized trials. The effectiveness of any antagonist depends not only on its ability to block the effects of the inflammatory mediators but also on its administration early enough in the course of the pancreatitis before pancreatic necrosis or organ dysfunction. PMID- 10470327 TI - Surgical management of necrotizing pancreatitis. AB - In necrotizing pancreatitis, surgical treatment is indicated in patients with infected necrosis. Conservative management should be favored if necrosis remains sterile and the patient responds to intensive care therapy. Different surgical techniques have been established during the past years, including conventional drainage, open and semiopen drainage, and closed management with postoperative continuous lavage of the lesser sac. For experienced physicians, these techniques provide comparable results, and none has been proved to be superior to the others. PMID- 10470328 TI - Progression from acute to chronic pancreatitis. A pathologist's view. AB - The available morphologic data on ACP are consistent with the view that ACP evolves from acute pancreatitis. How alcohol abuse triggers pancreatic injury and which factors are responsible for the progression to chronic pancreatitis remain to be clarified, however. PMID- 10470329 TI - Progression from acute to chronic pancreatitis: a physician's view. AB - Whether or not AP may progress to the chronic form is controversial. Equally debatable is whether AP caused by alcohol abuse develops in a chronically diseased gland or in a normal pancreas. As for the state of the gland, several postmortem studies have shown that AP may occur after acute alcohol abuse in the normal pancreas. As for progression from acute to chronic pancreatitis, many experimental studies have demonstrated signs of the chronic from of the disease in animals, but these signs were reversible. Some clinical studies have shown that alcohol-induced pancreatitis may progress to chronic pancreatitis. There are, however, presently no predictive parameters indicating when such a progression does or does not occur. PMID- 10470330 TI - Diagnosis of chronic pancreatitis. Is a gold standard necessary? AB - The question that serves as this article's title is rhetorical. Clinicians have diagnosed and managed chronic pancreatitis without a gold standard for decades and must continue to do so in the foreseeable future. Although clinicians have a much wider array of diagnostic tools available for the diagnosis of chronic pancreatitis, a single readily applied gold standard remains elusive. Diagnostic studies are rarely compared with a true gold standard--histopathology. Furthermore, even if a safe biopsy technique were available, it might fall short of a gold standard, given the patchy nature of early-stage chronic pancreatitis. Indeed, different stages of chronic pancreatitis require not only recognition of the different clinical presentations but also different levels of intensity of diagnostic testing to establish the diagnosis confidently. The diagnosis in most patients with chronic pancreatitis can be made confidently with a good clinical history and a limited number of currently available structural and functional tests. No single diagnostic study, functional or structural, suffices for all patients. It is also axiomatic that patients with intractable abdominal pain in whom early-stage chronic pancreatitis is suspected represent a challenge for clinicians partly because of this lack of a single, dependable gold standard. Perhaps we have reached the point at which further refinements of current tests of structure or function are not beneficial because increased sensitivity is countered by loss of specificity. We suggest that a new approach to developing a gold standard for the diagnosis of chronic pancreatitis is necessary. With advances in the understanding of the mediators of the inflammatory process, it may be possible to devise a test to assess the earliest events in this disease. PMID- 10470331 TI - Early and late onset in idiopathic and alcoholic chronic pancreatitis. Different clinical courses. AB - In summary, a prerequisite for the development of alcoholic pancreatitis would be the specific individual predisposition present in patients with late-onset idiopathic chronic pancreatitis. Furthermore, because the reported prevalence of chronic pancreatitis in patients with heavy alcohol consumption is markedly higher than the prevalence of late-onset idiopathic pancreatitis in the general population, the authors conclude that, in predisposed patients, alcohol consumption promotes the development of pancreatitis and accelerates the manifestation of symptoms and complications. This concept explains the observation that only a minority of severe alcoholics develop chronic pancreatitis. Conversely, in postmortem studies, a substantial proportion of older individuals without premortem evidence of pancreatic disease and no excessive alcohol history have pancreatic morphologic alterations resembling chronic pancreatitis. Thus, in the general population, a considerable number of asymptomatic "carriers," together with an undetected high prevalence of late onset idiopathic chronic pancreatitis, may exist. In these persons, alcohol consumption might amplify and accelerate preexisting asymptomatic idiopathic pancreatic damage. As a consequence, in a dose-dependent manner, alcohol may lead to an earlier onset of or induce clinically apparent pancreatitis in persons who otherwise might never have had symptoms during their lives. PMID- 10470332 TI - Palliation of pain in chronic pancreatitis. Use of enzymes. AB - According to the concept of negative feedback regulation of pancreatic enzyme secretion by proteases, treatment with pancreatic extracts has been proposed to lower pain in chronic pancreatitis by decreasing pancreatic duct pressure. The author, however, has demonstrated in healthy volunteers that intraduodenal application of porcine pancreatic extracts does not inhibit but rather stimulates pancreatic enzyme secretion. This is probably because of the high-protein content of porcine pancreatic extracts that may overwhelm a potential inhibitory effect of proteases. In a prospective placebo-controlled, double-blind multicenter study to investigate the effect of acid-protected porcine pancreatic extracts on pain in 43 patients with chronic pancreatitis, pain improved in most patients regardless of whether they started with placebo or verum. There was no significant difference between both treatment arms. In a meta-analysis, which included the author's study, six randomized, double-blind, placebo-controlled studies were evaluated. Statistical analysis demonstrated no benefit of the application of porcine pancreatic extracts to relieve pain in chronic pancreatitis. The author concluded that pancreatic extracts neither inhibit pancreatic enzyme secretion nor are they efficient in lowering pain in chronic pancreatitis. PMID- 10470333 TI - Palliation of pain in chronic pancreatitis. Use of neural blocks and neurotomy. AB - Effective management of the pain of chronic pancreatitis may require a multidisciplinary approach involving gastroenterologists, anesthesiologists, psychologists or counselors for chemical addiction (alcohol, narcotics), and surgeons. Viable approaches use pharmacologic analgesics with selected psychotropic medications, celiac plexus blocks, and possibly thoracoscopic splanchnic nerve transections. If these management techniques that preserve pancreatic parenchyma and function, fail, resective surgical therapy may be indicated. For most of these patients, all attempts at nonresective therapy should be exhausted before operative intervention. PMID- 10470334 TI - Interventional treatment of acute and chronic pancreatitis. Endoscopic procedures. AB - The role of therapeutic endoscopy in the treatment of acute and chronic pancreatitis has expanded dramatically over the past 10 years. Drainage of pseudocysts and even organized pancreatic necrosis when localized are becoming commonplace. Other areas in which therapeutic endoscopy has been shown to be efficacious include severe biliary pancreatitis, pancreatic duct disruptions, strictures, and obstructive calculi. Its role in the management of acute recurrent pancreatitis with presumed Oddi's sphincter dysfunction or pancreas divisum continues to be defined. The cost-effectiveness and minimally invasive nature of endoscopic therapy compared with surgery should ensure the continued development of these techniques. More controlled, prospective data are required. PMID- 10470336 TI - Antimalarial drug resistance and mortality in falciparum malaria. PMID- 10470335 TI - Surgical treatment of chronic pancreatitis and quality of life after operation. AB - In conclusion, surgical therapy in patients with chronic pancreatitis may be characterized as follows: 1. Independently, several investigators have found intraductal and intraparenchymatous hypertension in patients with chronic pancreatitis. Decompression of the ductal system as the main principle of surgical therapy achieves clinical pain relief in most patients with chronic pancreatitis. The precondition is a consequent drainage of the main pancreatic duct and tributary ducts of second and third order up to the prepapillary region. The presence of an inflammatory tumor in the head of the pancreas or ductal abnormalities in the prepapillary region or a pancreas divisum requires performance of an extended drainage operation (LPJ-LPHE) to achieve pain relief and an improved quality of life. An extended drainage operation effectively manages complications arising from adjacent organs, such as distal common bile duct stenosis, segmental duodenal stenosis, and internal pancreatic fistulas. The extent of decompression has to be tailored to the anatomic and morphologic situation of the patient. 2. In patients with chronic pancreatitis, the main pancreatic duct is usually dilated. A small duct (3-5 mm) is only small for the surgeon. For the sclerosing entity of chronic pancreatitis with a truly small duct, that is, less than 3 mm in diameter ("small duct disease"), a longitudinal V-shaped excision of the ventral pancreas, as opposed to left resection, provides a new perspective for a sufficient drainage. 3. In the presence of segmental portal hypertension, a simple or extended drainage operation does not result in a normalization of the portal venous blood flow; however, how often relevant upper gastrointestinal hemorrhage develops from segmental portal hypertension is unclear. Therefore, the clinical relevance of this special problem needs further evaluation. 4. Postoperative morbidity of LPJ-LPHE is significantly lower in comparison to resectional procedures, such as PD, PPPD, and DPRHP. A lower perioperative mortality rate is not justified anymore as a relevant criterion in favor of drainage procedures because resectional procedures are burdened by a minimal or no mortality in experienced centers; however, PD and PPPD are greatly hampered by a significantly decreased postoperative global quality of life as opposed to the LPJ-LPHE. This is reflected by a significantly lower rate of social and professional rehabilitation. 5. The incidence of exocrine and endocrine organ dysfunction is lower after LPJ-LPHE compared with PD or PPPD, but not compared with DPRHP. Preservation of the gastroduodenal passage and the continuity of the bile duct with its associated feedback mechanisms of exocrine pancreatic secretion and glucose metabolism seem to be responsible for this phenomenon. 6. An early surgical or endoscopic interventional drainage of the hypertensive pancreatic duct system possibly offers the chance to favorably manipulate the natural course of chronic pancreatitis with regard to a delayed onset of exocrine or endocrine insufficiency. 7. Late mortality reflects continued alcohol abuse rather than the effect of an operative procedure. PMID- 10470337 TI - The eosinophilic response and haematological recovery after treatment for Plasmodium falciparum malaria. AB - To examine a possible relationship between the immune response and haematological recovery after acute falciparum malaria, we followed peripheral blood eosinophil counts and haemoglobin concentrations for 4 weeks after starting effective treatment in 70 adult Thai patients. Eosinophils are induced by Th-2 cytokines as well as other stimuli. Eosinophil counts were elevated in only 8 (11%) of the subjects at presentation, but were increased in 65 (93%) by day 7. Eosinophil counts then decreased markedly by day 14, followed by a second increase until day 28. A significant positive correlation was found between peak eosinophil counts on day 7 and the haemoglobin concentration on day 28, both in 16 subjects without stool parasites (r = 0.65, P = 0.006) and in 54 patients with stool parasites (r = 0.32; P = 0.0019). These results suggest that a robust eosinophilic response shortly after completing antimalarial therapy predicts a good recovery from malaria-associated anaemia. PMID- 10470338 TI - Population structure of recrudescent Plasmodium falciparum isolates from western Uganda. AB - It has been proposed that polymorphisms of the Merozoite Surface Protein 1 and 2 (MSP1 and MSP2) and the Glutamate Rich Protein (GLURP) genes can be considered as genetic markers for the genotyping of field populations of Plasmodium falciparum. During a field study on in vivo drug resistance against chloroquine, sulphadoxine/pyrimethamine (S/P) and cotrimoxazole in West Uganda, sensitive and resistant isolates were collected from patients by fingerprick for genotyping. 59 (72.8%) of the 81 P. falciparum samples isolated at day 0 showed multiclonal infection with 2-7 clones. Among the isolates we investigated, presence of the allelic family MAD20 of MSP1 at day 0 was significantly (P = 0.0041) associated with decreased resistance to antimalarials. Use of this method in a field study on in vivo drug resistance demonstrates another potential application of genotyping as a tool for epidemiological investigations. PMID- 10470339 TI - Control of zoophilic malaria vectors by applying pyrethroid insecticides to cattle. AB - The principal method of malaria control in South and West Asia--indoor residual spraying--is extravagant with insecticide. A more efficient way of deploying insecticide might be to apply it on livestock since the malaria vectors in the region are highly zoophilic. A series of investigations assessed the potential of cattle treated with pyrethroid to control zoophilic mosquitoes. Permethrin, deltamethrin, and lambdacyhalothrin were applied to cattle using a sponge at concentrations of 0.5 g/m2, 0.025 g/m2, and 0.025 g/m2, respectively. 'Whole animal' insecticide bioassays that simulated natural mosquito-host contact were used to measure insecticidal and behavioural effects on wild host-seeking mosquitoes. The bioassays were performed on both sheltered and outdoor grazing animals for up to 45 days post-treatment. The possibility that treatments might cause diversion of host-seeking vectors from cattle to people nearby was also investigated. At the doses tested deltamethrin had the most pronounced and longest-lasting effect, reducing the proportion of bloodfed survivors by over 50% for the first two weeks. The majority of mosquitoes affected by the insecticide were killed before feeding. Insecticidal impact was generally lower on grazing than on sheltered animals. There was no diversion of host-seeking mosquitoes from treated cattle to nearby humans at any stage after treatment. The application of insecticide to livestock using a campaign approach may prove an effective means of controlling malaria in the region. At the doses tested deltamethrin is the most appropriate insecticide for this purpose. PMID- 10470340 TI - Retinal manifestations of HIV-1 and HIV-2 infections among hospital patients in The Gambia, west Africa. AB - BACKGROUND: In developed countries, 50-75% of AIDS patients develop retinal complications and about 20-40% acquire cytomegalavirus (CMV) retinitis. We conducted a cross-sectional survey to determine prevalence of these in The Gambia where both HIV-1 and HIV-2 infection are present and the prevalence of HIV-1 is rising. METHOD: All patients attending hospital whose percentage CD4+ cells (CD4%) was below 14, the level associated typically with an AIDS diagnosis, and one half of those whose CD4% was 14 or above were asked to join the study. Fifty six HIV-1, 52 HIV-2 and 12 dually infected patients were recruited. Photographs of the fundi were taken and interpreted independently. The findings were related to the patients' percentage CD4+ cells. RESULTS: The CD4% was < 14 in 40 patients and < 7 in 17 patients. Thirty-six patients were male. No cases of CMV retinitis were found. Four patients whose CD4% were 4, 5, 11 and 23 had cotton wool spots ranging in number from 1 to 14 for any one patient. The prevalence of cotton wool spots was 8% (95% CI, 0-16%) among patients with CD4% below 14 and 12% (95% CI, 0 27) among patients with CD4% below 7. One of the 4 patients had associated microaneurysm and blot haemorrhages typical of more advanced HIV microvasculopathy. CONCLUSION: CMV retinitis is less common in The Gambia than in developed countries. Non-infectious retinopathy may also be less common. PMID- 10470341 TI - Cortisol metabolism, cortisol sensitivity and the pathogenesis of leprosy reactions. AB - The concentration of cortisol in a tissue is regulated by a reversible enzyme 'shuttle' that can deactivate cortisol by converting it to cortisone, or activate cortisone by converting it to cortisol. The activity of this shuttle, and the direction in which it operates, is regulated by numerous factors including cytokines. This results in large swings in the effective cortisol concentration in sites of inflammation at different phases of an inflammatory response. Thus changes in local cortisol concentration can be largely independent of circulating cortisol levels. The relevant shuttle enzymes are present in skin, blood vessels and nervous tissue, and inhibition of the enzymes in skin enhances the local anti inflammatory effect of cortisol. We therefore suggest that changes in the activity or direction of action of the shuttle in leprosy lesions may predispose to reactions, requiring exogenous steroid supplements to regain control of the inflammation. PMID- 10470342 TI - Screening of army soldiers for Wuchereria bancrofti infection in the metropolitan Recife region, Brazil: implications for epidemiological surveillance. AB - Between 1989 and 1995, blood surveys were performed for Wuchereria bancrofti infection in several barracks of the Brazilian army in the metropolitan Recife region. For initial screening, 60 microliters of capillary blood were examined for microfilaria. All men who tested positive had microfilaria quantified by filtration of venous blood through a polycarbonate membrane. Of 23,773 men screened, 585 (2.5%) had microfilaria (mf). Microfilarial density ranged from < 1 8706 mf/ml of blood. Thirteen individuals had ultra-low microfilarial densities (1 mf/11 ml of blood). Characterization of 174 autochthonous cases made it possible to map 8 new districts in 4 cities within metropolitan Recife region where transmission of W. bancrofti was previously unknown. Routine screening of soldiers in the military may provide important surveillance data for national programmes to eliminate transmission of W. bancrofti. PMID- 10470343 TI - Is there a role for glycosuria testing in sub-Saharan Africa? AB - BACKGROUND: With increasing urbanization and westernization, rates of diabetes in sub-Saharan Africa (sSA) are likely to rise. Early detection and intervention plays an important role in delaying development of complications. In sSA in particular there is need for an affordable, reliable, safe, feasible test to avert human suffering and exhausting already stressed health facilities. METHODS: Data from two large community-based studies were used to assess the value of glycosuria testing in the detection of diabetes in adults in a sub-Saharan country. A first study (A) tested participants for glycosuria by dipstick; if positive, fasting capillary glucose was measured. A later study (B) measured glucose concentration in venous blood 2 h after a 75-g glucose load; if glycaemia was > or = 10 mmol/l, urine was tested for glycosuria. RESULTS: The positive predictive value of glycosuria for a diagnosis of diabetes (fasting glucose > or = 6.7 mmol/l) was 48%. Sensitivity was 64% (57% if a 2-h-value > or = 10 mmol/l was used as gold standard). Sensitivity was higher among overweight and/or hypertensive subjects, among elderly people in the urban area, and among subjects with higher blood glucose levels. Extrapolated specificity was 99.7%, and the likelihood ratio 190. CONCLUSIONS: Glycosuria testing can identify a considerable number of undiagnosed diabetic patients when specially targeted at high-risk groups (obese, hypertensive, or elderly people). Dipstick glycosuria testing is an appropriate, safe, feasible test for sSA, where the prevalence of diabetes is expected to increase considerably in the near future. PMID- 10470344 TI - Use of hospital data for Safe Motherhood programmes in south Kalimantan, Indonesia. AB - The evaluation of Safe Motherhood programmes has been hampered by difficulties in measuring the preferred outcomes of maternal mortality and morbidity. The need for adequate indicators has led researchers and programme managers alike to resort to indicators of utilization and quality of health services. In this study we assess the magnitude of four indicators of use of essential obstetric care (EOC) and one indicator of quality of care in health facilities in three districts in South Kalimantan, Indonesia. The general picture which emerges for South Kalimantan is that the use of obstetric services is low. Even in the more urban district of Banjar where facility-based coverage is highest, fewer than 14% of all deliveries take place in an EOC facility, 2% of expected births are admitted to such a facility with a major obstetric intervention (MOI), and 1% of expected births have an MOI for an absolute maternal indication. The use of facility-based EOC is consistently lower in Barito Kuala compared to the other districts, and the differences persist regardless of the indicators used. In this setting with low utilization rates, general rates of utilization of EOC facilities seem to be as satisfactory an indicator of relative access to EOC as more elaborate indicators specifying the reasons for admission. The inequalities in access to care revealed by the various indicators of use of EOC services may prove to be a more powerful stimulus for change than the widely reported and highly inaccurate accounts of the high levels of maternal mortality. PMID- 10470345 TI - Viewpoint: the neglect of stool microscopy for intestinal parasites and possible solutions. PMID- 10470346 TI - 'Unexplained death' from malaria tropica mistaken for viral haemorrhagic fever. PMID- 10470347 TI - The prehistory of psychiatry in Ethiopia. AB - Ethiopian psychiatry has changed considerably during the last few years with an increasing number of Ethiopian doctors and nurses trained in psychiatry. In the paper is given an outline of the history of psychiatry in Ethiopia from 1965 onwards. Important improvements in the Amanuel Mental Hospital in Addis Ababa which for long was the only psychiatric facility in the country, the establishment of the Department of Psychiatry at the Medical Faculty of the University of Addis Ababa and the introduction of psychiatry in the curriculum of the medical faculty are important steps. Recently training of nurses and doctors in psychiatry has led to the establishment of psychiatric clinics in 26 district hospitals throughout the country staffed with psychiatric nurses supervised by psychiatrists from Amanuel Hospital in Addis Ababa. The development of psychiatric research in Ethiopia is also outlined. PMID- 10470348 TI - The prevalence and socio-demographic correlates of mental distress in Addis Ababa, Ethiopia. AB - The study was conducted in Addis Ababa, the capital city of Ethiopia, between January and March of 1994, to estimate the prevalence of mental distress using the Self Reporting Questionnaire (SRQ). The study population of 10,203 was selected from the entire city by a cluster sampling method proportionate to size. Five percent of the study population reported at least 2 of the 4 SRQ items for probable psychosis. By use of the 20-item SRQ for assessing mental distress, and using a cut-off level of at least 6 out of 20 items, 11.7% of the study population was categorized as having mental distress. Multivariate logistic analysis showed that women had a statistically significant 62% increased risk of having mental distress than men. There was a statistically significant trend of increasing risk with increasing age. There was also a significant trend of reduced risk with increasing educational attainment and family size. Employment was also inversely associated with risk: adjusted Odds Ratio and 95% confidence interval, OR (95% CI) = 0.81 (0.69, 0.95). Family history of mental illnesses was positively associated with risk: OR (95% CI) = 3.98 (3.18, 4.98). Marital status, ethnicity and religion were not significantly associated with risk of mental distress. PMID- 10470349 TI - Major mental disorders in Addis Ababa, Ethiopia. I. Schizophrenia, schizoaffective and cognitive disorders. AB - We report here on a survey conducted in Addis Ababa between September and December of 1994 to estimate the prevalence and socio-demographic correlates of schizophrenia, schizoaffective, and severe cognitive disorders. An Amharic version of the CIDI was used to collect data from a random community sample of 1420 individuals aged 15 and above. Weighted lifetime and one-month prevalence were 0.4% and 0.3%, for schizophrenia, and 0.5% and 0.4% for schizoaffective disorders, respectively. The prevalence of severe cognitive disorders was found to be 2.6%. Multivariate logistic analysis showed that the risk of lifetime psychosis (schizophrenia, schizoaffective, bipolar disorders, and depressive episodes with psychotic symptoms) was significantly higher in the age group 25-44 years compared to the reference age group of 15-24 years: adjusted Odds Ratio and 95% confidence interval, OR (95% CI) = 2.41 (1.01, 5.74); it was also significantly higher in the never married group compared to those married: OR (95% CI) = 3.01 (1.08, 8.42). The risk was significantly lower in the employed compared to the unemployed: OR (95% CI) = 0.42 (0.18, 1.00). There was a statistically significant trend of increasing risk of severe cognitive disorders with increasing age. The trend was in the reverse direction for educational status, the risk decreasing significantly with increasing educational attainment. PMID- 10470350 TI - Major mental disorders in Addis Ababa, Ethiopia. II. Affective disorders. AB - This report examines the prevalence and socio-demographic correlates of affective disorders based on a survey conducted in Addis Ababa between September and December of 1994. An Amharic version of the CIDI was used to collect data from a random community sample of 1420 individuals aged 15 and above. The lifetime prevalence for specific affective disorders was as follows: bipolar disorders 0.3%, depressive episodes 2.7%, recurrent depressive episodes 0.2%, and persistent mood disorders 1.6%. The weighted lifetime prevalence of affective disorders was 5.0% (women 7.7% and men 3.2%). One-month prevalence was 3.8% (women 5.9% and men 2.3%). After adjusting for several potential confounders, the risk of affective disorders was only 29% higher in women compared to men. This difference in risk was not statistically significant. Age was also not associated with risk of affective disorders. On the other hand, education was associated with the risk of disorder, the risk decreasing with increasing educational attainment. This inverse trend was statistically significant (P for trend = 0.02). The risk was also 37% lower in the employed than the unemployed: Odds Ratio (OR), 95% confidence interval (95% CI) = 0.63 (0.39, 1.01). There were no statistically significant associations between affective disorders and marital status or ethnicity. PMID- 10470351 TI - Major mental disorders in Addis Ababa, Ethiopia. III. Neurotic and somatoform disorders. AB - This report presents estimates of the lifetime and point (one-month) prevalence of phobic anxiety, dissociative, other anxiety and somatoform disorders based on a survey conducted in Addis Ababa between September and December of 1994. An Amharic version of the CIDI was used to collect data from a random community sample of 1420 individuals aged 15 and above. Phobic anxiety disorders were found to be the most common of all anxiety disorders, with lifetime prevalence of 4.8%. The lifetime prevalence for dissociative disorders was 0.8% and for other anxiety disorders it was 2.7%. The prevalence of somatoform disorders was 3.1%. The lifetime prevalence for all the above neurotic and somatoform disorders was 10.8%. Point prevalence estimates (percent) for the disorders were as follows: phobic anxiety disorders, 4.4; other anxiety disorders, 1.2; dissociative disorders, 0.4; and somatoform disorders, 2.5. After adjusting for several potential confounders, women had over a two-fold risk of having neurotic and somatoform disorders compared to men, odds ratio (OR), 95% confidence interval (CI) = 2.38 (1.78, 3.12). Age was also inversely associated with the disorders. Those aged 60 and above had a 60% lower risk compared to those aged 15-24 years, OR (95% CI): 0.41 (0.20, 0.81). The 25% decreased risk for the employed, compared to the unemployed, was also not statistically significant. Education, marital status, and ethnicity were not significantly associated with neurotic and somatoform disorders. PMID- 10470352 TI - The epidemiology of alcohol dependence and problem drinking in Addis Ababa, Ethiopia. AB - We describe here a prevalence study of alcohol dependence and problem drinking in a representative sample of 10,203 adults in Addis Ababa. At the first stage the study employed a 4-scale screening instrument (CAGE) and at the second stage the Composite International Diagnostic Interview (CIDI) was used to identify cases of alcohol dependence. Of the total population, 2.7% responded positively to at least 2 of the 4 CAGE items, fulfilling the definition of problem drinking. By use of the CIDI, the weighted lifetime and one-month prevalence of alcohol dependence was 1.0% and 0.8%, respectively. It occurred almost exclusively among men. The prevalence of problem drinking increased with increasing age. The trend was statistically significant (P for trend = 0.03). On the other hand, there was a statistically significant negative trend in the association with educational level, use decreasing with increasing educational attainment (P for trend = 0.0006). There was also a statistically significant 39% increased risk of alcohol use with employment. The association with ethnicity was not statistically significant. Only sex was significantly associated with alcohol dependence. Women had a 84% less risk of becoming dependent compared to men. PMID- 10470353 TI - Suicide attempts and ideation among adults in Addis Ababa, Ethiopia. AB - Attempted suicide is one of the major risk factors for completed suicide, is associated with psychiatric disorders, and is also a potentially fatal event. We describe here a study of suicide attempts and ideation in a representative sample of 10,203 adults in Addis Ababa. The study was conducted between January and March 1994. The prevalence of current suicidal ideation was 2.7%. Lifetime prevalence of suicidal attempt was 0.9%. Most of the attempts (66%) occurred when subjects were under 25 years of age. Hanging was the preferred method for men and poisonings for women. There were no significant associations between suicide attempt and sex, ethnicity or religion. Current suicidal ideation was more common in men than women, odds ratio (95% confidence interval), OR (95% CI): 0.67 (0.48, 0.93). There was a statistically significant trend of decreasing risk of ideation with increasing age and educational attainment. There was a 68% decrease in the risk of ideation in the 60 years of age and above group compared to the 15-24 year group: OR (95% CI): 0.32 (0.16, 0.62). Marital status, ethnicity, and religion were not associated with suicidal ideation. PMID- 10470354 TI - How are mental disorders seen and where is help sought in a rural Ethiopian community? A key informant study in Butajira, Ethiopia. AB - One hundred key informants were interviewed about their awareness, attitudes and practices regarding mental illness using the Key Informant Questionnaire developed by WHO. Case vignettes of seven common neuropsychiatric disorders were presented to the key informants. Informants' awareness about these disorders and help-seeking practices for mental and physical symptoms or conditions were assessed. An additional question on the prototype symptoms of mental disorders was also posed. Among the presented seven conditions, epilepsy was perceived as the most common condition and major depression was regarded as the least common one. Schizophrenia was judged as the most severe problem, and mental retardation was considered the second most severe condition. Talkativeness, aggression and strange behaviour were the most frequently perceived prototype symptoms of mental illness. Traditional treatment methods were preferred more often for treating symptoms of mental disorders and modern medicine was preferred more often for treating physical diseases or symptoms. Findings of this study are similar to other studies conducted in socio-culturally different communities. Working in close connection with traditional healers would give the primary health care worker a better opportunity to gain acceptance from the community and modify certain harmful practices. PMID- 10470355 TI - The prevalence and socio-demographic correlates of mental distress in Butajira, Ethiopia. AB - A cross-sectional survey was conducted on 10,468 rural and semi-urban adults in an Ethiopian district using the Self Reporting Questionnaire (SRQ) to detect the prevalence of mental distress and its association with socio-demographic risk factors. Fifty-eight per cent of the study population were women, 74% were Muslim, 79% were illiterate. Those experiencing 11 or more symptoms out of the 20 SRQ items were considered as having mental distress. Accordingly, the prevalence of mental distress was 17%, which is comparable with the previous hospital-based studies in Ethiopia and elsewhere. However, it was higher than the previous community-based studies in Ethiopia. Mental distress was more prevalent among women. Part of the explanation was that women in the study population were older and that they were more often widowed or divorced, which were factors associated with mental distress. Illiteracy, which was more common among women and older individuals, was also independently associated with mental distress. PMID- 10470357 TI - Awareness and attitudes of a rural Ethiopian community toward suicidal behaviour. A key informant study in Butajira, Ethiopia. AB - One hundred key informants were interviewed regarding their awareness and attitudes toward suicidal behaviour. Eighty-eight informants were male, 58 were Muslim and 42 were Christian. Informants on average, claimed to know more persons who had completed suicide than those who had attempted suicide. Almost all informants mentioned more than one cause for suicide. Of these, frustration was the most frequently mentioned cause. Most informants believed that suicide attempters are cruel, feared and not trustworthy. Their attitude toward suicide completers was expressed as condemned sinners, do not deserve funeral ceremony, and should be buried separately from others. Christians gave importance to the funeral issue more than did the Muslims. Generally, the attitudes of informants were punitive and disapproving. PMID- 10470356 TI - Major mental disorders in Butajira, southern Ethiopia. AB - Previous studies conducted in Ethiopia lack information on the prevalence of specific mental disorders in rural communities. The lifetime and one-month prevalence of specific ICD-10 mental disorders and their associated socio demographic factors were determined using the translated Amharic version of the Composite International Diagnostic Interview (CIDI) in a rural population. A total of 501 community subjects selected from a predominantly rural district by stratified random sampling were interviewed by non-clinician interviewers. The weighted aggregate lifetime prevalence of psychiatric morbidity was 31.8% (26.7% when substance dependence was not included). The most frequent specific diagnoses were: dissociative disorders (6.3%), mood disorders (6.2%), somatoform disorders (5.9%), and anxiety disorders (5.7%). After adjustment in a multivariate logistic model, female sex was shown to have a statistically significant association with mood disorders (Odds Ratio, OR = 3.84, 95% CI: 1.90, 7.73) and somatoform disorders (OR = 2.30, 95% CI: 1.13, 4.60). Severe cognitive and mood disorders were significantly associated with being elderly, i.e. 60 or more years of age (OR = 7.71, 95% CI: 1.58, 7.53; and OR = 3.68, 95% CI = 1.36, 9.95, respectively). Khat dependence was associated with being Muslim and with earning a low income. (OR = 3.5, 95% CI: 1.02, 11.98; and OR = 0.32, 95% CI: 0.10, 0.96, respectively). It is concluded that psychiatric morbidity is a major public health problem in the rural community. PMID- 10470358 TI - Suicide attempts among adults in Butajira, Ethiopia. AB - In a cross-sectional survey, 10,468 adults of a rural and semi-urban community were interviewed to determine lifetime suicide attempts. Among the study population, 58% were female, 74.4% were Muslim and 79.3% had had no formal education. The majority of the population were in the age group 25-59 years. Lifetime suicide attempt was reported by 3.2% (n = 332) of the study population. Of these, 63% (n = 208) were women. The most frequent age of attempt was between 15 and 24 years and the frequency of attempt decreased with increasing age. Hanging and poisoning were the most frequently reported methods of attempting suicide. Marital or family conflict was the most frequently reported cause for attempting suicide and most of those who reported this cause were women (Chi square = 17.42; P < 0.001). Men were significantly more likely to use hanging to attempt suicide than women (Chi-square = 8.21; P < 0.001). Among Christians 3.9% had a lifetime suicide attempt compared to 2.9% among Muslims (Chi-square = 6.15; P < 0.05). People who currently had mental distress and problem drinking reported lifetime suicide attempt more often than others. PMID- 10470359 TI - The epidemiology of problem drinking in Butajira, Ethiopia. AB - In order to determine the prevalence and socio-demographic correlates of problem drinking, a total of 10,468 persons aged 15 and above, most residing in a rural district, were interviewed using the CAGE questionnaire as an important element of a general mental health survey. Twenty-three per cent of the respondents admitted that they currently drank alcohol. The prevalence of alcohol drinking was 15% for women and 36% for men. Among those who drank, 16% met the criterion for problem drinking as defined by two or more positive responses to the CAGE. The overall prevalence for problem drinking was found to be 3.7%. Stratified analysis for sex showed that Christian religion, male sex, being ethnically non Gurage, and smoking were strongly associated with problem drinking in both sexes. Marital status, mental distress and income were found to be associated factors with problem drinking only in men. PMID- 10470360 TI - The prevalence and socio-demographic correlates of khat chewing in Butajira, Ethiopia. AB - A house-to-house survey was carried out in a rural Ethiopian community to determine the prevalence and socio-demographic correlates of khat use. A total of 10,468 adults were interviewed. Of these, 58% were female, and 74% were Muslim. More than half of the study population (55.7%) reported lifetime khat chewing experience and the prevalence of current use was 50%. Among current chewers, 17.4% reported taking khat on a daily basis; 16.1% of these were male and 3.4% were female. Various reasons were given for chewing khat; 80% of the chewers used it to gain a good level of concentration for prayer. Muslim religion, smoking and high educational level showed strong association with daily khat chewing. PMID- 10470361 TI - Childhood behavioural disorders in Ambo district, western Ethiopia. I. Prevalence estimates. AB - The study was conducted between September 1994 and May 1995 in Ambo district, western Ethiopia. The prevalence of childhood behavioural disorder in children was found to be 17.7%. Behavioural disorder was found to be more common in boys than in girls. The prevalence increased with age. The most frequent symptoms reported were headache and nervousness. The least prevalent symptom was stealing things from home. As age increased, the risk of behavioural disorder increased. The increase in risk was statistically significant in the 15-year-old group when compared to the age group 5-7 years (adjusted Odds Ratio, OR = 1.89, 95% confidence interval, CI: 1.08-2.85). Childhood mental disorder was statistically significantly associated with parental age and with parental marital status. Children whose parents were < or = 24 years old had a higher risk of having mental disorders (OR: 2.03, 95% CI: 1.30-3.16) compared to those children whose parents were in the 45+ age group. Children whose parents were categorized as unmarried, divorced, separated, or widowed had a higher risk of having behavioural disorders (OR: 2.22, 95% CI: 1.70-2.91) than children whose parents were married. There was a statistically significant association between parental psychoneurosis and children's behavioural disorders; children whose mothers had psychoneuroses were at a higher risk of having behavioural disorders as compared to those whose mothers had no psychoneurosis (OR: 1.78, 95% CI: 1.34-2.35). PMID- 10470362 TI - Childhood behavioural disorders in Ambo district, western Ethiopia. II. Validation of the RQC. AB - We report here on a study conducted to measure the validity of an Amharic version of the Reporting Questionnaire for Children (RQC), that was used in a survey of childhood behavioural disorders in a predominantly rural district in western Ethiopia. Mothers of 196 children aged 5-15 years, who were initially interviewed by the RQC were re-interviewed by a psychiatrist who was unaware of the RQC status of these children. The re-interview was conducted using a DSM IV checklist. The study showed that a cut-off point of one or more positive responses to any of the 10 questions on the RQC maximized sensitivity (87.5%) and specificity (65%). The discriminatory power of each item was also computed, and the item dealing with wetting/soiling oneself was found to have the highest ability to identify cases from non-cases. The item on abnormal speech was found to have the least discriminating power. PMID- 10470363 TI - Triapine (3-aminopyridine-2-carboxaldehyde thiosemicarbazone; 3-AP): an inhibitor of ribonucleotide reductase with antineoplastic activity. AB - The enzyme RR catalyzes the conversion of ribonucleoside diphosphates to their deoxyribonucleotide counterparts. RR is critical for the generation of the cytosine, adenine, and guanine deoxyribonucleotide 5'-triphosphate building blocks of DNA, which are present in cells as exceedingly small intracellular pools. Therefore, interference with the function of RR might well result in an agent with significant antineoplastic activity, particularly against rapidly proliferating tumor cells. HUr is the only inhibitor of RR in clinical usage; this agent, however, is a relatively poor inhibitor of the enzyme and has a short serum half-life. Consequently, HUr is a relatively weak anticancer agent. In an effort to develop a more potent inhibitor of RR with utility as an anticancer agent, we have synthesized 3-AP and demonstrated (a) potent inhibition of L1210 leukemia cells in vitro, (b) curative capacity for mice bearing the L1210 leukemia, (c) marked inhibition of RR, and (d) sensitivity of HUr-resistant cells to 3-AP. These findings collectively demonstrate the clinical potential of 3-AP as an antineoplastic agent. PMID- 10470364 TI - Causes and consequences of acidic pH in tumors: a magnetic resonance study. AB - A consequence of metabolism in any tissue is the formation of hydrogen ions, which are actively transported out of the cell. However, although most solid tumors maintain their intracellular pH (pHi) within a narrow range to provide a favorable environment for various intracellular activities, their extracellular pH (pHe) is on average about 0.2 pH units more acid. It is important to understand the relationship between tumor metabolism and pH, and how it differs from that of normal tissue and to ask the question: How does an understanding of pH and tumor metabolism affect strategies for therapeutic approaches? Although, in vitro, isolated cell experiments have shown positive correlations between pHi and pHe the relationship is complex and somewhat dependent on experimental conditions. Measurement of pHi in solid tumors by non-invasive Magnetic Resonance Spectroscopy (MRS) has been possible for some time now and recently several specific markers for measuring pHe have become available. As a result we have been able to study the relationship between pHi and pHe in vivo in several different solid tumor types. In only one tumor type (HT29 xenografts) was there a significant correlation between pHi and pHe; in 3 other tumor types (RIF-1 in mice, GH3 prolactinomas and H9618a in rats) there was no correlation. A significant correlation between pHi and NTP/Pi ratios was seen across all tumor types. Theoretical considerations of causes of tumor acidity, hypotheses to explain extracellular acidity and the possibility that low pHe might be an intrinsic feature of the tumor phenotype and not merely the consequence of metabolic activity have been discussed. In addition the consquences for concepts of treatment based on pH are considered. PMID- 10470365 TI - Phosphatidylinositol 3-kinase translocation to the nucleus is an early event in the interleukin-1 signalling mechanism in human osteosarcoma Saos-2 cells. AB - Interleukin 1 (IL-1) is a proinflammatory cytokine which can elicit proliferative, differentiative, or metabolic responses. The molecular mechanisms by which IL-1 signals are transduced from the plasma membrane to the nucleus, although extensively studied, have not been completely elucidated. We previously demonstrated that human osteosarcoma Saos-2 cells incubated with IL-1 presented a rapid and transient increase of phospholipase C activity exclusively at the nuclear level. Moreover, we presented evidence that not only the canonical inositol lipid signalling pathway was involved, but also the D3-phosphorylated lipids generated by phosphatidylinositol 3-kinase (PI 3-kinase) were affected. The results of this study indicate that in Saos-2 cells PI 3-kinase is recruited and activated by IL-1 receptor I (IL-1RI) through binding of the SH2 domains to the consensus sequence on the C-terminal tail of the receptor, and that Tyr-479 is essential for PI 3-kinase activation. Moreover, IL-1 treatment triggers PI 3 kinase translocation to the nucleus; this event is rapid and transient in cells expressing high levels of IL-1RI (Saos-2/IL-1R) as well as in untransfected cells, although to a lesser extent. The data, based on immunochemical and immunocytochemical quantitative methods, indicate that PI 3-kinase translocation to the nucleus depends on PI 3-kinase activation. In fact, inactivation by two independent mechanisms, addition of specific PI 3-kinase inhibitors, or overexpression of a mutant form of IL-1RI, resulted in a substantial inhibition of PI 3-kinase translocation to the nucleus. These data suggest that PI 3-kinase recruitment by the activated receptor is a limiting step in PI 3-kinase activation and nuclear translocation. This early event in the IL-1 signalling mechanisms confirms that D3 inositides, as well as canonical inositides produced by nuclear phospholipase C isoforms, are involved in this pathway of activation of transcription factors. PMID- 10470366 TI - Amplification of signal transduction capacity and down-regulation by drugs. AB - Recent work in this Laboratory showed increased activity of PI 4-kinase, PIP kinase and PLC in various cancer cells, indicating a stepped-up capacity for signal transduction. This elevated potential was paralleled with increased concentration of the end product of signal transduction, IP3. Current investigations showed that in normal cells the activities of the specific phosphatases (which degrade PIP2 and PIP and oppose those of the synthetic enzymes) were 4 to 5 orders of magnitude higher than those of the synthetic kinases. In hepatoma cells the specific phosphatase activities markedly decreased. Thus, in cancer cells the marked elevations in activities of the synthetic enzymes were opposed by a reduction in the activities of the degradative specific phosphatases. This enzymic imbalance is responsible, in part at least, for the elevated capacity of signal transduction and IP3 concentration. Since the enzymic activities measured were proportionate with time elapsed and amount of enzyme added, the alterations in activities should reflect changes in enzyme amounts. These alterations indicate a reprogramming of gene expression which should confer selective advantages to the cancer cells, marking out the elevated synthetic enzyme activities as potentially sensitive targets for drug treatment. We showed earlier that tiazofurin, which curtailed the biosynthesis of enzymes with short half-lives such as PI and PIP kinases, down-regulated signal transduction and brought down IP3 concentration. Quercetin and genistein chiefly inhibited PI-4 kinase and PIP kinase, respectively, and as a result reduced IP3 concentration in cancer cells. Current studies reveal that tiazofurin with quercetin, tiazofurin with genistein, and quercetin with genistein were synergistic in killing human cancer cells and in reducing signal transduction activity. In estrogen receptor-negative MDA-MB-435 human breast carcinoma cells which have elevated signal transduction activity, tamoxifen caused IC50S for growth inhibition and cytotoxicity of 12 and 0.7 microM, respectively. When tiazofurin was added to breast carcinoma cells, followed 12 hr later by tamoxifen, synergism was observed in growth inhibition, in clonogenic assays and in the reduction of IP3 concentration. The synergistic action of tiazofurin and tamoxifen and the other synergistic drug interactions outlined above may have implications in the clinical treatment of neoplasias. PMID- 10470367 TI - Regulation of homocysteine metabolism. AB - We have used a combination of in vivo and in vitro techniques to measure factors regulating homocysteine metabolism and the plasma concentration of this atherogenic amino acid. The germane findings include: 1. Homocysteine metabolism in rat kidney proceeds predominantly through the transsulfuration pathway, whose enzymes are enriched within the proximal cells of kidney tubules. Furthermore, the rat kidney possesses significant reserve capacity to handle both acute and chronic elevations in plasma homocysteine concentrations. 2. Plasma homocysteine concentrations are lower in diabetic rats. Insulin administration corrects this perturbation. Therefore, insulin and/or one of its counter-regulatory hormones affects homocysteine metabolism, possibly through an increased flux in the hepatic transsulfuration pathway. In support of these data, glucagon administration to rats produced similar results. Further support was provided by studies with isolated rat hepatocytes, from which homocysteine export was reduced when incubated in the presence of glucagon. PMID- 10470368 TI - Homocysteine metabolism in cardiovascular cells and tissues: implications for hyperhomocysteinemia and cardiovascular disease. AB - We have determined the activity and protein levels of CBS in a number of cardiovascular cells and tissues by direct enzyme assay and Western blot analysis, respectively. We have also determined the activity of BHMT in these same tissues and cells and have come to the conclusion that neither enzyme is expressed. This results suggests that in the human cardiovascular system homocysteine metabolism is limited to the remethylation pathway catalyzed by MS. Thus, hyperhomocysteinemia in conjunction with a limited metabolic capacity for homocysteine in the cardiovascular system could result in cellular dysfunction. PMID- 10470369 TI - Lack of competition of substrates for P-glycoprotein in MCF-7 breast cancer cells overexpressing MDR1. AB - The MCF-7/Adr cells overexpress MDR-1 which contributes to the drug-resistant phenotype. Our studies show: 1. The retention of daunomycin in the MCF-7/Adr cells relates to a temperature-dependent and energy-dependent process. 2. The MCF 7/Adr cells retain less rhodamine-123 than the parental MCF-7 cells. 3. The MCF 7/Adr cells retain less daunomycin than the parental MCF-7 cells as measured by mean fluorescence or radioactive daunomycin. 4. Cyclosporin A and verapamil effectively block the effluxes of rhodamine-123 and daunomycin from the MCF-7/Adr cells. 5. On short-term incubation, 2-deoxyglucose lowers the NTP levels to a greater extent than sodium azide, showing the importance of glycolysis in the MCF 7 cell lines. 6. Although the MCF-7/Adr cells show cross-resistance to VP-16, actinomycin D and vinblastine, these drugs do not compete with daunomycin for the efflux pump. 7. These data suggest that either there must be multiple MDR-1 pumps that differ in substrate specificity or that there are distinct substrate sites on MDR-1. PMID- 10470370 TI - Genomic amplification of the human DHFR/MSH3 locus remodels mismatch recognition and repair activities. AB - Mismatch recognition in human cells is mediated by two heterodimers, MutS alpha and MutS beta. MutS alpha appears to shoulder primary responsibility for mismatch correction during replication, based on its relative abundance and ability to recognize a broad spectrum of base-base and base-insertion mismatches. Because MutS alpha and MutS beta share a common component, MSH2, conditions that influence the expression or degradation of MSH3 or MSH6 can redistribute the profile of mismatch recognition and repair. MSH3 is linked by a shared promoter with DHFR, connecting two pathways with key roles in DNA metabolism. In a classic example of gene amplification, the DHFR (and MSH3) locus can become amplified to several hundred copies in the presence of methotrexate. Under these conditions, MutS beta forms at the expense of MutS alpha, and the mutation rate in these tumor cells rises more than 100-fold. The implications for cancer chemotherapy include a potential increase in mutability when tumors are treated with methotrexate, which could increase the frequency of subsequent mutations that influence the tumor's drug sensitivity or aggressiveness. Because processing certain types of DNA damage by the mismatch repair pathway has also been implicated in tumor sensitivity to agents such as cisplatin, changes in expression at the DHFR/MSH3 locus may have further relevance to the outcome of multi-drug treatment regimens. PMID- 10470371 TI - Screening for inducers of apoptosis in apoptosis-resistant human carcinoma cells. AB - Many anticancer agents are known to induce apoptosis in cultured cells, but human solid tumor cells are often resistant to apoptosis induction. Human pancreatic adenocarcinoma AsPC-1 cells were found to be resistant to apoptosis. So, we screened microbial culture filtrates and synthetic chemicals for their ability to induce apoptosis in AsPC-1 cells. Polyoxypeptin A was isolated from a Streptomyces culture broth as a potent inducer of apoptosis. Its cyclic hexadepsipeptide structure contains a simple but hitherto unreported amino acid, (2S,3R)-3-hydroxy-3-methylproline. Polyoxypeptin A induced early cell death, apoptotic morphology, and internucleosomal DNA fragmentation in AsPC-1 cells at low concentrations. Polyoxypeptin A can induce caspase 3 activation in human T cell leukemia Jurkat cells but not in AsPC-1 cells. The mechanism of apoptosis in AsPC-1 cells has not been elucidated yet. Less toxic derivatives of polyoxypeptin A are being prepared. A macrocyclic lactam called BE-14106 was also isolated from Streptomyces as an inducer of apoptosis in AsPC-1 cells. Histidine-pyridine histidine-3 (HPH-3) is an oxygen-activating ligand derived from the structure of bleomycin, and it also induced apoptosis in AsPC-1 cells. Induction of apoptosis by HPH-3 was inhibited by zinc and copper ions, indicating that chelation with ferrous ion is responsible for induction of apoptosis, as is chelation by bleomycin to cleave DNA. Bleomycin A2 and its portion having no DNA-binding region, glycopeptide-3(GP-3), did not induce apoptosis in AsPC-1 cells. Thus, among the actions of bleomycin-related compounds, the induction of apoptosis is a unique characteristic of HPH-3. PMID- 10470372 TI - Potassium is a critical regulator of apoptotic enzymes in vitro and in vivo. AB - Apoptosis, or programmed cell death, is a fundamental biological process involved in many physiological and pathological phenomena. This process is predominantly catabolic in which cellular macromolecules are broken down by distinct enzymes to be later recycled in healthy cells. These enzymes are arranged in an elaborate cascade that serves to both propagate and amplify a death signal as well as process bulk macromolecules and inhibit repair systems. One of the best characterized enzyme systems involved in apoptosis is the activation of a nuclease(s) that degrades the genome into discrete oligonucleosomal fragments, clearly committing a cell to death. Using two different in vitro models, we have shown that K+ directly inhibits this/these nuclease(s) with complete inhibition observed at cellular concentrations of K+ found in non-dying cells (150 mM). These data suggest that K+ concentrations in living cells suppress apoptotic nuclease activity. One of the nucleases involved in apoptotic DNA degradation (NUC18/Cyclophilin) has been purified in our laboratory and found to be inhibited by similar concentrations of K+. Upstream of DNA fragmentation a specific class of proteases, termed caspases, are activated which propagate an apoptotic signal and lead to downstream events such as DNA fragmentation. Using an in vitro model of caspase activation, we also observed that activation of this enzyme is also completely inhibited by normal physiological K+ levels, suggesting that K+ levels in non-dying cells suppress multiple portions of the apoptotic enzyme cascade. These results suggest that K+ concentrations may decrease in cells undergoing apoptosis and both physical and fluorescence techniques document an intracellular K+ concentration of 35 mM in apoptotic cells. This loss of K+ also accounts for changes in cell volume (cell shrinkage) that are universally associated with apoptosis. Measurements of K+ concentration in shrunken and non-shrunken cells demonstrated that only shrunken cells contain a reduced concentration of intracellular K+. Importantly, both the nuclease and caspase activity were found exclusively in this shrunken population providing an absolute correlation between the activity of these enzymes and low K+ levels within a cell. Finally, suppressing K+ efflux in whole cells prevents the activation of these enzymes whereas enhancing the efflux of this ion facilitates enzymatic activity. These results suggest a direct cause and effect relationship between the level of K+ and the activity of apoptotic enzymes. Taken together we have shown a critical and novel mechanism that regulates apoptosis by a direct effect on the apoptotic enzymes. PMID- 10470373 TI - Expression of enzymes of covalent protein modification during regulated and dysregulated proliferation of mammary epithelial cells: PKA, PKC and NMT. AB - Three proteins are functionally interlinked in the targeting of protein phosphorylation catalyzed by the C-subunit of PKA: PKA itself, AKAPs and NMT. Furthermore, in a variety of biological contexts, mechanisms exist whereby PKA and PKC are able to modulate the activity of one another. We have investigated the expression and subcellular distribution of these proteins in two models of mammary cell proliferation and differentiation--the normal rat mammary gland during pregnancy and lactation and human breast tissue before and after malignant transformation. Modulation of PKA does not acutely affect activity or sub cellular distribution of PKC in mammary acini, nor does modulation of PKC acutely affect PKA activity or subcellular distribution. Therefore, the co-ordinate expression of these two protein kinases in normal and cancerous mammary epithelial cells and the greater basal activation level of them both accompanying increased mitogenic activity, which we have reported, does not result from short term cross-talk between them. Although basal and total levels of PKA diminish in rodent mammary epithelial cells during the transition from proliferative to secretory functional mode, the level of expression of AKAPs increases. The expression of two apparently mammary-specific and mostly membrane-associated AKAPs is tightly linked to the onset and maintenance of differentiated function in rat mammary tissue. Paradoxically, the probable analogues of these two AKAPs in human mammary tissue are hyperexpressed when normal epithelial cells transform to a cancer phenotype--conventionally regarded as a process involving a degree of dedifferentiation. Mammary AKAP hyperexpression in breast cancers is accompanied by increases in the levels of total and basal PKA. One mechanism whereby NMT is targeted to membranes, via interaction with ribosomal proteins, has recently been elucidated. Our data support the contention that the localization of NMT is an important variable in the regulation of cellular proliferation, but they do not characterize the mechanisms whereby the differential targeting of NMT is achieved. As yet we lack a full tool-kit with which to examine NMT either to draw firm conclusions regarding the identity of particular isoforms found in particular sub-cellular locations or to define the relationships between these different molecular variants. However, it is technically possible to transfect cells with inducible NMT expression constructs engineered in such a way that the recombinant, catalytically competent, NMT that they encode is targeted either to membranes or to cytosol: an exploration of the effects of such transfections on cellular proliferation would afford a critical test of the mechanistic involvement of NMT in the control of mitogenesis. PMID- 10470374 TI - The regulation of acetyl-CoA carboxylase--a potential target for the action of hypolipidemic agents. AB - ACC exists as two major isoforms ACC1 or ACC alpha, and ACC2 or ACC beta, and there is evidence that they play separate roles in the production of malonyl-CoA for fatty acid synthesis and the control of mitochondrial beta-oxidation, respectively. ACC alpha can be regulated at the level of gene expression, allosteric regulation of the enzyme, and reversible phosphorylation by AMP-PK. Emerging lines of research suggest that similar mechanisms of regulation exist for ACC beta. Its inactivation in heart and skeletal muscle through phosphorylation by AMP-PK is becoming well-established. ACC is an important target of certain hypolipidemic drugs such as the fibrates. This is not simply because ACC alpha inhibition decreases the synthesis of a lipid component of VLDL because fatty acids synthesized de novo in liver are not always major contributors to VLDL lipid (158); it is also because ACC beta inhibition leads to a decrease in malonyl-CoA levels and the disinhibition of fatty acid oxidation. Partitioning fatty acids towards oxidation and away from esterification is an important aspect of the lipid-lowering effects of fibrates. Fibrates could use any of the mechanisms of ACC regulation to decrease activity. They could repress ACC gene expression through the activation of PPAR alpha, and fibroyl-CoA esters could inhibit ACC allosterically just as TOFyl-CoA does. However, we have demonstrated a rapid inactivation of ACC in cultured rat hepatocytes by gemfibrozil that is mediated by activation of AMP-PK and the subsequent phosphorylation of ACC. The end result is the inhibition of hepatic fatty acid synthesis and a possible activation of beta-oxidation as evidenced by the increased production of ketone bodies. The mechanism through which fibrates activate the AMP-PK cascade, the role of PPAR alpha, the physiological responses of biosynthesis and oxidation and the use of these mechanisms by other hypolipidemic agents are areas of ongoing investigation. PMID- 10470375 TI - Export pumps for anionic conjugates encoded by MRP genes. AB - Several members of the multidrug resistance protein (MRP) family mediate the ATP dependent transport of amphiphilic anions across membranes. The substrate specificity of recombinant human MRP1 has been most extensively defined by use of inside-out membrane vesicles. Substrates include the glutathione S-conjugate leukotriene C4, 17 beta-glucuronosyl estradiol, glucuronosyl bilirubin, glutathione disulfide, in addition to the fluorescent lipophilic anion Fluo-3. These substances are also substrates for the apical isoform MRP2, also termed canalicular multispecific organic anion transporter, cMOAT, which shares only 49% amino acid sequence identity with MRP1. The K(m) of leukotriene C4 for MRP2 is 10 fold higher than for MRP1, and the K(m) of 17 beta-glucuronosyl estradiol is 4.8 fold higher for MRP2 than for recombinant human MRP1. Human as well as rat MRP2 confer multidrug resistance to polarized MDCKII cells permanently expressing the recombinant glycoprotein in their apical plasma membrane. Resistance of cells transfected with human and rat MRP2 to etoposide was enhanced 5-fold and 3.8 fold, and resistance to vincristine was enhanced 2.3-fold and 6.0-fold, respectively. Conjugate-transporting members of the MRP family with a related sequence and a similar function have been detected recently. In addition to several MRP isoforms (MRP1-6) and orthologs in mammals (human, rat, rabbit, mouse), MRP family members have been identified in the nematode Caenorhabditis elegans, in the yeast Saccharomyces cerevisiae, and in the plant Arabidopsis thaliana. These conjugate export pumps of the MRP family play a widespread role in detoxification, drug resistance, and, because of the role in the export of glutathione disulfide by MRP1 and MRP2, in the defense against oxidative stress. PMID- 10470376 TI - Study of the functional share of lysosomal cathepsins by the development of specific inhibitors. AB - To analyze the functional share of individual cathepsins, we developed powerful and specific inhibitors for individual cathepsins using computer graphics of substrate binding pockets based on X-ray crystallography. These new inhibitors were named CLIK group. Epoxy succinate peptide derivatives, CLIK-066, 088, 112, 121, 148, 181, 185 and 187, are typical specific inhibitors for cathepsin L. Aldehyde derivatives CLIK-060 and CLIK-164 showed specific inhibition against cathepsin S and cathepsin K, respectively. We found that pyridoxal phosphate (PLP), a coenzyme form of vitamin B6, inhibits all cathepsins and also new artificially synthesized pyridoxal derivatives, CLIK-071 and -072, in which the phosphate esters of PLP were replaced by propionic acid, exhibited strong inhibition for cathepsins. Furthermore, CLIK-071 was easy to incorporate into cells and showed powerful inhibition for intracellular cathepsins. Using these selective inhibitors, the allotment of individual cathepsin functions in cells has been studied as follows. Cathepsin L and/or K participate in bone resorption based on bone type-1 collagen degradation and the L-type protease inhibitors suppressed the bone resorption. Cathepsins B and S participate in antigen presentations based on antigen processing and invariant chain degradation, respectively. Also cathepsin L participates in cell apoptosis mediated by caspase III activation. PMID- 10470377 TI - Folylpoly-gamma-glutamate synthetase: generation of isozymes and the role in one carbon metabolism and antifolate cytotoxicity. AB - A single human gene encodes both mitochondrial and cytosolic isoforms of the enzyme. The major mRNA species in human cells encodes the mitochondrial isoform but alternate translation initiation at a downstream in-frame ATG also generates the cytosolic isoform. Cytosolic FPGS may also be generated by use of alternate transcription initiation start sites 3' to the start ATG of the mitochondrial FPGS. Three additional human FPGS mRNAs differing in exon 1 have been identified. One of these is a major species in HEP-G2 cells and other tissue culture cells, and can encode a protein lacking the first 8 amino acids of cytosolic FPGS. A protein of the predicted size is observed in coupled transcription/translation systems. However, expression of this protein in E. coli does not generate an active enzyme. Mutagenesis studies indicate that Tyr-3 of the missing N terminal residues is required for enzyme activity. The major cellular folate pools are in the cytosol and mitochondria and FPGS activity is normally distributed in both compartments. Mitochondrial FPGS activity is required for mitochondrial folate accumulation, and cells lacking this isozyme are auxotrophic for glycine. Overexpression of cytosolic FPGS does not complement the lack of mitochondrial activity. Cells expressing FPGS activity solely in the mitochondria are glycine prototrophs, but also possess cytosolic folylpolyglutamates and are prototrophic for thymidine and purines, products of cytosolic one carbon metabolism. Although cytosolic folylpolyglutamates cannot enter the mitochondrion, mitochondrial folylpolyglutamates are released intact into the cytosolic compartment. Cellular accumulation of some antifolates and their cytotoxic efficacy is highly responsive to the level of FPGS activity. Polyglutamylation of methotrexate (MTX) has little affect on its affinity for dihydrofolate reductase, its target enzyme, but does affect the cellular accumulation of the drug. The sensitivity of model cells, expressing a range of FPGS activities similar to that observed in leukemia blasts, to MTX varied over four orders of magnitude. MTX toxicity was dependent on cytosolic FPGS activity as this drug does not enter the mitochondria, and cells expressing very high levels of FPGS solely in the mitochondria were resistant to MTX. The cytotoxic efficacy of other folate antagonists that are transported into the mitochondria was enhanced by mitochondrial FPGS activity, even when their loci of inhibition was a cytosolic enzyme. Mitochondrial metabolism of these drugs increased cytosolic drug levels. Compartmentalization of antifolate metabolism has to be considered in evaluating mechanisms for increased drug cytotoxicity and for the development of acquired resistance to these agents. PMID- 10470378 TI - Membrane sidedness of biosynthetic pathways involved in the production of lysophosphatidic acid. AB - Lysophosphatidic acid (LPA) is a novel phospholipid mediator with diverse biological activities such as smooth muscle contraction, and proliferative effects or modifications of cytoskeleton. Activated blood platelets are the best identified source, explaining accumulation of LPA in serum upon blood coagulation. However, the metabolic pathways responsible for LPA synthesis are still poorly known. Using a model of human erythrocytes treated with the calcium ionophore A23187, we have shown that type II secretory phospholipase A2 (sPLA2) is able to produce LPA by hydrolyzing phosphatidic acid exposed on the cell surface after phospholipid scrambling. A similar mechanism does not appear to occur in platelets, where inhibitors of sPLA2 or genetic lack of the enzyme do not modify LPA production. However, this does not definitely eliminate the possibility that LPA is also produced in platelets in the external leaflet of the membrane by other phospholipases, which have to be better characterized. PMID- 10470379 TI - Inositides in the nucleus: further developments on phospholipase C beta 1 signalling during erythroid differentiation and IGF-I induced mitogenesis. AB - Inositol lipids originally shown to be metabolized in the cytosol have been detected also in the nucleus, where they are both synthesized and hydrolyzed. In the case of erythroid differentiation of murine erythroleukemia cells (Friend cells) it has been previously shown that PLC beta 1, which is the major nuclear PLC, undergoes down-regulation upon treatment with DMSO or tiazofurin which act as differentiative agents. On the contrary, i.e., during IGF-I induced mitogenesis, it has been shown that PLC beta 1 is rapidly activated and this event is essential for the onset of DNA synthesis. Even though its key role in cell growth has been shown, both the mechanism by which nuclear PLC beta 1 is activated and the direct relationship with erythroid differentiation are still unknown. We have addressed the question if PLC beta 1 expression and activity in the nucleus are directly related or not to the establishment of the differentiated state and we have checked the two main ways of activation, i.e., via G-protein or via phosphorylation, in order to establish whether nuclear PLC beta 1 is regulated the same way as the one at the plasma membrane or not. The data reported here show that nuclear PLC beta 1 is responsible for a continuous recycling of Friend cells, acting as a negative regulator of differentiation and that its activation is dependent on the phosphorylation state. PMID- 10470380 TI - Lipid kinases are novel effectors of the GTPase Rac1. AB - We have found that a complex consisting of a type I PIPK and a DGK associates with the GTPase Rac1. Binding of the lipid kinase complex is through the C terminus of Rac. Complex formation is augmented in the presence of specific phospholipids. The complex also associates with Rho GDI, through Rac. Based on the role of PtdIns-4,5-P2 in regulating proteins that influence actin structures we propose that the Rac-associated lipid kinase complex functions to generate locally high concentrations of PtdIns-4,5-P2 in a Rac-dependent manner. There are many possible roles PtdIns-4,5-P2 might play. A likely role is binding to barbed end actin capping proteins. This would release the capping protein, providing free barbed ends for actin polymerization. Uncapping would occur at the membrane so that additional actin polymerization would result in membrane protrusions and lamellapodia, in a Brownian ratchet model. It is also possible that PtdIns-4,5-P2 has other roles, such as promoting the release of G actin from profilin or promoting the cross-linking of actin or its anchorage to the plasma membrane. Studies are currently underway to determine the role of this lipid kinase complex in Rac signaling and actin regulation in vivo. PMID- 10470381 TI - Enzymatic rationale and preclinical support for a potent protein kinase C beta inhibitor in cancer therapy. AB - The macrocyclic bisindolylmaleimide, LY333531, selectively inhibits protein kinase C beta 1 and beta 2 isoforms with an approximate IC50 of 5 nanomolar. The efficacy of LY333531 administered alone and in combination with cytotoxic cancer therapies in models of non-small cell lung carcinoma and brain tumors was determined in vivo. In the Lewis lung carcinoma, administration of LY333531 enhanced the activity of paclitaxel and fractionated radiation and, to a lesser degree, carboplatin and gemcitabine. In the human T98G glioblastoma multiforme xenograft, the addition of LY333531 to treatment with carmustine (BCNU) resulted in enhanced tumor response in a nodule grown subcutaneously and increased life span in animals bearing an intracranial tumor from 37 days in the control animals to 64 days in the BCNU treated animals, and to 104 days in the LY333531 plus BCNU treated animals with 4 out of 5 animals being long-term survivors. PMID- 10470382 TI - Expression of human herpesvirus-8 (HHV-8) encoded pathogenic genes in Kaposi's sarcoma (KS) primary lesions. AB - Transcription of six different HHV-8 specific mRNAs was examined in early- and late-stage KS primary lesions. Expression of the latency-associated T0.7 mRNA and of VP23 mRNA which is a specific marker of lytic/productive infection suggested that HHV-8 is secondarily recruited into the KS lesions by productively infected monocytes, macrophages. From these cells HHV-8 is transmitted to the KS spindle cells, which are latently infected. v-BCL-2, v-MCP-1 and v-IL-6 were not expressed in latently infected KS spindle cells, therefore the impact of these factors in KS pathogenesis appears to be low. By contrast, v-Cyclin D was highly expressed in almost all latently infected spindle cells and may therefore be an important factor triggering progression of late-stage KS lesions. PMID- 10470383 TI - Identification and cloning of a novel isoform of mouse secretory leukocyte protease inhibitor, mSLPI-beta, overexpressed in murine leukemias and a highly liver metastatic tumor, IMC-HA1 cells. AB - Several genes showing transcriptional alteration in a highly liver metastatic murine carcinoma cell line, IMC-HA1, were identified by mRNA differential display system. Among them, a gene identical to mSLPI was isolated as mSLPI-alpha and beta. They were produced through an alternative splicing. Their full-length cDNA sequences were determined, and their expression in various murine tumors and normal tissues was analysed. The deduced translation product of mSLPI-alpha showed 59% identity to hSLPI. Although mSLPI-beta had the same 103-amino-acid sequence from the carboxyl terminus, the amino terminus showed hydrophilicity opposite mSLPI-alpha or hSLPI. The mSLPI-alpha was expressed ubiquitously in various tumor cell lines. Interestingly, however, mSLPI-beta expression was only observed in P388 and L1210 leukemias and IMC-HA1 cells, and in lower amounts in three normal tissues (thymus, lung and spleen), suggesting that mSLPI, and in particular the unusual splicing product, mSLPI-beta, plays a specific role in these cells, including malignant processes of tumor cells. PMID- 10470384 TI - Cell signaling by protein tyrosine phosphorylation. AB - The above data, and others not described herein, indicate the following: First, that phosphatases are not scavenger enzymes, simply there to remove the phosphate groups introduced by the kinases. They cannot be viewed simply as providing an 'off' switch in an 'on/off' kinase/phosphatase system. Kinases and phosphatases do not carry out one-way and opposing reactions. The same enzyme, depending on where it localizes within the cell, or the molecule with which it might interact, can serve either as a positive or negative determinant in defining cell behavior. In many instances, it can act synergistically with the kinases to enhance the phosphorylation reaction. Second, the factors that determine whether a phosphatase would enhance or oppose a kinase reaction would seem to depend less on its state of activity than on its subcellular localization. This would suggest that if one wanted to call upon it to control transformation, one should try to tamper with its localization segments or whatever binding proteins it might be attached to--rather than with its catalytic domains. Displacement of these enzymes from where they are meant to bind would seem a more promising approach than trying to modulate their catalytic activity. Finally, their architectural features are so basically different from those of the kinases, with receptor tyrosine phosphatases displaying all the structural characteristics of cell adhesion molecules, that they must also have a mission of their own in cell development, survival and death, quite apart from that of the kinases. PMID- 10470385 TI - Maximillian Neu and the first anaesthetic rotameter. PMID- 10470386 TI - Effect of positive end-expiratory pressure on left and right ventricular diastolic filling assessed by transoesophageal Doppler echocardiography. AB - The effect of positive end-expiratory pressure (PEEP) on left and right ventricular diastolic filling dynamics was assessed by transmitral and transtricuspid flow patterns. Using transoesophageal Doppler echocardiography in fourteen ASA physical status 1 female patients, the following measurements were performed at baseline (0 cm H2O PEEP) and at 5, 10, 15, and 20 cm H2O PEEP: 1. peak velocity of early filling (peak E velocity), 2. peak velocity of atrial contraction (peak A velocity), 3. the ratio of the peak E to A velocity (peak E/A velocity ratio), 4. isovolumic relaxation time (IRT), 5. acceleration half-time (AHT), 6. deceleration half-time (DHT) of early filling, and 7. end-diastolic and end-systolic areas of both ventricles. Increasing PEEP progressively deceased peak E velocity of both ventricles. In contrast, peak A velocity did not change and the peak E/A velocity ratio decreased significantly with PEEP. IRT and AHTs remained unchanged, but DHTs of both ventricles increased following PEEP. End diastolic and end-systolic areas of both ventricles decreased gradually and significantly with PEEP. It is concluded that PEEP was associated with decreased preload as well as reduced compliance of both ventricles, which was considered to contribute to the changes in diastolic ventricular filling. PMID- 10470387 TI - Propofol-thiopentone admixture-hypnotic dose, pain on injection and effect on blood pressure. AB - This study examined some pharmacodynamic characteristics of two admixtures of propofol and thiopentone. Ninety unpremedicated ASA 1 or 2 patients were group randomized to receive, in a double-blinded manner, one of the following mixtures for induction of anaesthesia: Group P50: propofol 1% 10 ml/thiopentone 2.5% 10 ml; Group P75: propofol 1% 15 ml/thiopentone 2.5% 5 ml; Group P100: propofol 1% 20 ml/lignocaine 1% 4 ml. An additional 30 randomized but unblinded patients from the same patient cohort received thiopentone 2.5% to provide predictive dose data for groups P50 and P75. Haemodynamic data were collected pre- and post-induction. The required induction dose of both mixtures of propofol and thiopentone found an additive rather than a synergistic interaction with no significant difference between predicted and observed dose. Thiopentone resulted in significantly more rapid induction of anaesthesia than propofol/lignocaine or propofol/thiopentone. The addition of thiopentone to propofol was found to be as efficacious as the mixing of lignocaine with propofol in reducing pain on injection. The fall in systolic blood pressure was significantly less in group P50 compared with groups P75 or P100. Admixture of thiopentone with propofol results in an additive hypnotic effect, a reduction in pain of injection (comparable with addition of lignocaine) and a reduced hypotensive response compared to propofol injection alone during induction. PMID- 10470388 TI - Extravascular lung water and acute respiratory distress syndrome--oxygenation and outcome. AB - We studied eleven consecutive patients to assess the influence of extravascular lung water on clinical outcome. All patients were mechanically ventilated using a standardized protocol. Inspired oxygen concentration was adjusted to an initial target PaO2 of greater than 8.0 kPa (60.8 mmHg). All patients received inhaled nitric oxide (NO) at a concentration of 20 ppm. Extravascular lung water index (EVLWI) was measured by a dual indicator technique (COLD Z-021 monitoring system, Pulsion, Munich, Germany). Patients were managed with fluids and inotropes according to a standard protocol. Median age was 45 (range 27-60) years, mean APACHE II score on admission 31 (range 17-36), duration of mechanical ventilation 15 (range 6-28) days, mean admission Murray lung injury score 2.5 (range 2-3) and admission EVLWI 20.8 (range 8.7 to 54.7) ml.kg-1. The only variables independently predictive of PaO2/FiO2 ratio were serum albumin (B = 1.7 +/- 1.61) and EVLWI (B = -2.1 +/- 0.47), r2 = 0.33, P < 0.0001. In severe ARDS, (PaO2/FiO2 < 150 mmHg), mean EVLWI was 24.4 (22.4 to 26.4, 95% confidence intervals) ml.kg-1 compared with 15.1 (12.2 to 18.0) ml.kg-1 during moderate ARDS (P < 0.001). Serum albumin likewise differed, 29.4 (27.6 to 31.2) vs 35.1 (31.8 to 38.4) g.l-1, P < 0.005. PAOP was higher during periods of poor oxygenation, 12.7 (11.9 to 13.5) vs 9.3 (7.9 to 10.7) mmHg, P < 0.001. The four survivors had greater initial EVLWI than non-survivors, 31 (24.1 to 37.9) vs 20.7 (16.0 to 25.4) ml.kg-1, P = 0.034 and showed a greater reduction in lung water, 15.2 (9.3 to 21.1) vs 5.4 (2.1 to 8.7) ml.kg-1, P = 0.013. PMID- 10470389 TI - Induction of anaesthesia in patients with coronary artery disease: a comparison between sevoflurane-remifentanil and fentanyl-etomidate. AB - In a prospective, randomized study, sevoflurane-remifentanil (Group SR) was compared with fentanyl-etomidate (Group FE) for induction of anaesthesia in patients with ischaemic heart disease. Cardiovascular stability, heart rate, mean arterial pressure, rate pressure product, rescue medications and associated myocardial ischaemia were measured. For Group SR (n = 20), anaesthesia was induced with vital capacity breaths of sevoflurane 5% in oxygen. After loss of consciousness, the inspired sevoflurane was reduced to 3% and remifentanil was administered as a 0.5 microgram.kg-1 bolus over 90 seconds (0.33 microgram.kg 1.min-1) followed by a 0.025 microgram.kg-1.min-1 infusion. After intubation, the inspired sevoflurane was reduced to 2%. For Group FE (n = 20), anaesthesia was induced with fentanyl 10.5 micrograms.kg-1 and etomidate 0.2 mg.kg-1 given 60 seconds later. Isoflurane 1% in oxygen was administered after loss of consciousness. Both groups received rocuronium and the trachea was intubated two minutes later. Sevoflurane gaseous induction was smooth, with cardiovascular stability comparable to fentanyl-etomidate. Significantly more patients in Group SR (P < 0.05) were on beta-blocking medication, and, overall, the HR and RPP was lower pre-intubation in Group SR. Remifentanil administration was associated with severe bradycardia in three patients and asystole in a fourth. All four patients were on beta-blocking medication and three of the four were on diltiazem. The study was terminated due to the high incidence of bradycardic/asystolic complications in Group SR. PMID- 10470390 TI - Cisatracurium and atracurium as antigens. AB - Seventy-five consecutive patients referred to an anaesthetic allergy clinic were intradermally tested with atracurium and cisatracurium. With the exception of one patient the results were identical, suggesting that allergy to either drug is associated with allergy to the stereoisomer. For skin testing for allergy to neuromuscular blocking drugs it is only necessary to use either atracurium or cisatracurium and cisatracurium is the preferred drug. PMID- 10470391 TI - The antiemetic and dysphoric effects of droperidol in the day surgery patient. AB - The incidence of side-effects of two doses of droperidol used as a prophylactic antiemetic were compared (10 vs 20 micrograms/kg). Two hundred and twenty-eight women for day case laparoscopy were recruited. Pain and nausea scores were collected in the recovery area and in the Day Surgery Unit prior to discharge. A telephone follow-up questionnaire was administered. Incidences of anxiety, restlessness and dysphoric reactions were similar in both groups (29.2% vs 29.0%). The incidence of postoperative vomiting varied from 4.4 to 7.7%. There were no significant differences between the two groups when nausea scores, incidences of vomiting and pain after discharge were compared. An unexpected finding was the significantly higher incidence of pain in the 10 micrograms/kg group. We conclude that there is no advantage in lowering the dose of droperidol below 20 micrograms/kg in this group of patients. PMID- 10470392 TI - The impact of an acute pain service on postoperative pain management. AB - An audit of postoperative pain management was conducted before and after the introduction of an Acute Pain Service (APS) run entirely by medical staff. The ability of patients to complete two pain-scoring systems, a verbal rating score (VRS) and a numerical rating score (NRS, where 0 = no pain, 10 = worst pain) was compared. We surveyed 605 adults 24 hours postoperatively. For major operations, the incidence of "severe" or "unbearable" pain at rest (VRS) over the first 24 hours decreased from 18.1% before to 3.5% after the APS (P = 0.0002) and severe/unbearable pain with movement decreased from 50% to 31% (P = 0.0037). The average NRS pain scores fell from 4.65 to 3.37 at rest (P < 0.0001) and from 6.77 to 6.19 with movement (P = 0.046). The incidence of severe/unbearable pain at rest with patient-controlled analgesia (PCA) decreased from 19.7% to 3.2% after the APS (P = 0.0012) and with movement from 51.3% to 35.1% (P = 0.049). For epidural analgesia, severe/unbearable pain at rest was 18.8% prior to the APS and 4.4% after (P = 0.14), and with movement was 43.8% before and 19.1% after (P = 0.079). The NRS pain-scoring system was unsuitable for Aboriginal or Torres Strait Islander patients. Patient satisfaction was high both before and after the introduction of the APS, and was an unreliable indicator of effective pain relief. We conclude that an APS can improve postoperative pain control with PCA and epidural analgesia. PMID- 10470393 TI - Undernutrition in children--effect on vecuronium induced neuromuscular blockade. AB - Sixty children aged one to 12 years requiring anaesthesia including a muscle relaxant were assessed for their nutritional status based on simple anthropometric and biochemical parameters. They were allocated to one of four groups: normal nutrition, mild, moderate or severe malnutrition. The neuromuscular effects of vecuronium bromide 0.1 mg/kg were studied by recording evoked responses to train of four (TOF) nerve stimulation using an accelerograph. In the above nutritional groups, time to onset of 25% depression of T1 was 0.8, 1.4, 1.3 and 2.1 minutes respectively. Maximal depression of TOF response was seen at 2.2, 3.2, 3.7 and 8.4 minutes. The duration of action of the initial dose was 26.5, 24.0, 17.7 and 13.3 minutes and the mean duration of action of top-up doses was 16.2, 14.9, 11.2 and 8.9 minutes respectively. Reversal time with neostigmine 0.05 mg/kg was not significantly different in the four groups. These results demonstrate a statistically significant delay in onset and shortening of the duration of action of vecuronium in the undernourished groups compared with the normal nutrition group when vecuronium is administered to children on a milligram per kilogram basis. PMID- 10470394 TI - Patient referral and transportation to a regional tertiary ICU: patient demographics, severity of illness and outcome comparison with non-transported patients. AB - A descriptive analysis and comparison of critically ill transported patients with non-transported patients will assist in selecting the appropriate referral and transportation process and subsequent incorporation into the critical care services of receiving hospitals. A retrospective review of transported and non transported patients admitted to the same Intensive Care Unit was conducted. Patient demographics, disease categories, source of admission to ICU, APACHE II scores, predicted and actual hospital mortality, hospital and ICU length of stay were examined. Of all ICU admissions, 16% were transported. Transported patients had a different case mix, significantly higher severity of illness measures, mortality and length of ICU stay. Observed mortality of transported patients with sepsis, gastrointestinal disease or bleeding, intracranial haemorrhage and post respiratory arrest was less than predicted whilst those with neurological disease, post cardiac arrest and overdose had a higher than predicted mortality. PMID- 10470395 TI - A survey of Australasian obstetric anaesthesia audit. AB - In order to develop a minimal obstetric anaesthesia dataset based on current Australasian clinical audit best practice, we carried out a postal survey of 69 Australasian anaesthetic departments covering an obstetric service. We asked about data being collected, specifically concerning the high risk obstetric patient, epidural analgesia and postoperative anaesthetic review. Examples of any data collection forms were requested. Of the 66 responses, 35 departments (53%) were not collecting any audit data. Twenty-six of the 31 departments (84%) performing obstetric anaesthesia audit responded to our follow-up telephone survey. Eighteen departments believed that there had been an improvement in patient care as a result of their audit and 13 felt that the benefits outweighed the costs involved. However, only six departments (9%) had performed an audit cycle. The importance of feedback to patients or hospital staff and the incidence of post dural puncture headache (PDPH) were cited by some as priorities for obstetric anaesthesia audit. There was however no consistency as to what data should be collected. Many responses suggested a perceived need to collect clinical data without knowing what to do with it. Our survey has highlighted confusion between three distinct objectives; a dataset for obstetric anaesthesia record keeping, data required for continuing patient management in hospital and, a specific minimal dataset for clinical audit purposes. We conclude that current Australasian obstetric anaesthesia audit strategies are inadequate to develop a minimal dataset for cost-effective clinical audit. PMID- 10470396 TI - Vancomycin resistance in gram-positive cocci--old bugs, new tricks, confusing terminology. PMID- 10470397 TI - Failure of exhalation during ventilation with the Drager Oxylog 2000 ventilator. AB - We describe three cases in which there was failure of exhalation during mechanical ventilation with the Drager Oxylog 2000 ventilator. Two potential mechanisms for this failure were identified, and were then experimentally reproduced on a test lung. First, a manufacturing fault in the silicone diaphragm in the ventilation valve was discovered. When a small hole in the diaphragm was incompletely perforated, a possible flap valve was formed, limiting exhalation. Second, failure of the one-way rubber disc valve to seat properly in its housing also prevented exhalation. These two previously unreported modes of ventilator failure have been identified. Careful checking of the ventilator circuit before each use is required. PMID- 10470398 TI - Extracorporeal circulatory support in near-fatal flecainide overdose. AB - Flecainide, a class 1c antiarrhythmic, has a high mortality associated with significant overdose. We report the case of a 20-year-old female who took approximately 4 grams of flecainide and a small amount of paracetamol as an impulsive gesture. Circulatory failure unresponsive to pacing, inotropes and sodium bicarbonate was successfully treated with cardiopulmonary bypass (CPB). Resolution of her myocardial failure occurred over 24 hours and she was weaned from CPB 30 hours after its initiation. Coagulopathy and intravascular haemolysis were apparent during bypass and necessitated substantial use of blood products. Ischaemic renal dysfunction manifested early in her admission and required haemodiafiltration. Despite a prolonged period of unresponsiveness and pupillary dilatation during resuscitation and CPB she made a full recovery. We believe this is the first reported case of flecainide overdose, requiring extracorporeal circulatory support, not resulting in neurological deficit. PMID- 10470399 TI - Failed intubation in the intensive care unit managed with laryngeal mask airway and percutaneous tracheostomy. AB - We report the management of failed intubation in a critically ill, hypoxic and catabolic patient with sepsis and acute lung injury. Insertion of a laryngeal mask airway restored ventilation and corrected hypoxia. As the laryngeal mask provides only a temporary airway, it was essential to secure the airway by percutaneous tracheostomy to initiate mechanical ventilation. PMID- 10470400 TI - Use of inhaled nitric oxide in pulmonary embolism. AB - Acute massive pulmonary embolism carries a high mortality with the majority of deaths occurring during the early phase. We describe a case of massive pulmonary embolism resulting in severe cardiovascular collapse and cardiac arrest which was treated successfully with inhaled nitric oxide. PMID- 10470401 TI - Management of a pregnant patient with airway obstruction secondary to goitre. AB - A case of airway obstruction in advanced pregnancy is presented. The patient was successfully managed with an awake fibreoptic intubation performed orally followed by a caesarean section and thyroidectomy as a combined procedure. On resection, a thyroid gland weighing 370 g was removed. The patient made an uneventful recovery. PMID- 10470402 TI - Gas embolism produced by hydrogen peroxide abscess irrigation in an infant. AB - The production of gaseous oxygen when hydrogen peroxide interacts with tissue is a well-known phenomenon that has been reported as a rare cause of gas embolism. We present the case of an 11-month-old infant who sustained an immediate cardiorespiratory arrest following the use of this agent during a minor surgical procedure. Clinical features, radiological findings and the rapid response to resuscitation were strongly suggestive of major gas embolism. Adverse effects of inappropriate use of hydrogen peroxide, and the diagnosis and management of these problems are discussed. PMID- 10470403 TI - Anaesthesia in the prone position for impalement injury. AB - A 22-year-old man was brought to the Emergency Department in the prone position, having been impaled in the right buttock with a large pitchfork. He was in excruciating pain and nauseated; attempts to move the patient or the pitchfork caused severe pain. Caudal blockade was carried out in the prone position, without moving the patient, and proved to be very efficacious. The pitchfork was then removed painlessly, allowing us to turn the patient supine. A conventional induction of general anaesthesia was then made possible. PMID- 10470404 TI - Prone ventilation. PMID- 10470405 TI - Inadvertent cannulation of an aberrant radial artery and intra-arterial injection of midazolam. PMID- 10470406 TI - Central venous catheter related infection. PMID- 10470407 TI - Application of the upgraded image superimposition system (SIAS) to the assessment of sperm kinematics. AB - Dedicated software for semiautomatic objective motility assessment, based on image superimposition (SIAS), was upgraded in order to allow automatic reading of sperm tracks whilst keeping the advantages of visual check instead of the digital images of classic computer-aided sperm analysis (CASA) systems. The new system is very fast and accurate and allows an excellent reading of sperm motility in standard semen analysis. Moreover, using this system, an attempt was made to define numerically straight-line velocity (VSL), curvilinear velocity (VCL) and linearity (LIN) in each WHO sperm motility class. At 21 frames sec-1, ROC curves analysis determined the following: Class 1 (typical of WHO grade 'a'): VSL > or = 23 microns sec-1 and LIN > or = 0.58; class 2 (typical of WHO grade 'b'): VSL > 10 and < 23 microns sec-1 and LIN > or = 0.58; class 3: VSL > 10 microns sec-1 and LIN < 0.58 (this additional class was added to differentiate nonstraight progressive motility from classes 1 and 2); class 4 (typical of WHO grade 'c'): VSL < or = 10 microns sec-1. The numerical definition of sperm motility classes may contribute towards standardization in the objective evaluation of sperm kinematics. PMID- 10470408 TI - In vitro effects of peroxynitrite on human spermatozoa. AB - In the present study, the in vitro effects of peroxynitrite on sperm motility, lipid peroxidation and sulfhydryl content were examined. Sperm percentage motility and movement characteristics were assessed by a computer-assisted system. Lipid peroxidation was measured by determining malondialdehyde levels using the thiobarbituric acid (TBA) method. Sperm sulfhydryl content was measured by a spectrophotometric method based on reduction of 5,5'-dithiobis-(2 nitrobenzoic acid) by sulfhydryl groups. Percentage motility, movement characteristics and sulfhydryl content decreased significantly in peroxynitrite treated samples compared to decomposed peroxynitrite-treated samples. Lipid peroxidation in peroxynitrite-treated samples was significantly higher than in decomposed peroxynitrite-treated samples. These results indicate that peroxynitrite anion may cause sperm dysfunction through lipid peroxidation stimulation and total SH depletion. PMID- 10470409 TI - A new point mutation of the androgen receptor gene in a patient with partial androgen resistance and severe oligozoospermia. AB - Mutations of the androgen receptor gene in genetic males cause a variety of androgen insensitivity syndromes varying from female phenotype through intersexuality to male phenotype with infertility. The identification of a missense mutation in the steroid-binding domain in an infertile male with mild features of androgen insensitivity is reported here. PMID- 10470410 TI - Mast cells in testicular biopsies of infertile men with 'mixed atrophy' of seminiferous tubules. AB - Mast cells in the bilateral testicular biopsies of 30 patients with a 'mixed atrophy' of seminiferous tubules were analysed. Seven biopsies from vasectomized patients served as controls. With regard to their characteristic location within testicular tissue, two groups of mast cells could be distinguished, in both control and infertile patients: 'interstitial' mast cells (located between Leydig and other interstitial cells as well as in the vicinity of blood vessels) and 'peritubular' mast cells (located in the close proximity of the tubular lamina propria or incorporated in the lamina propria itself). Morphometric data indicated a significant increase in the number and volume of mast cells in infertile patients when compared with controls. In the biopsies of infertile patients that were analysed both 'interstitial' and 'peritubular' mast cells showed a significant increase in their number and volume, although it appeared that 'peritubular' mast cells increased at a higher rate than 'interstitial' mast cells. A significant negative correlation was found between the following variables: volume and number of mast cells, testis volume and the status of spermatogenesis evaluated by Johnsen's scoring. It was concluded that the increased presence of mast cells is closely associated with an impairment of spermatogenesis. PMID- 10470411 TI - Effects of extracellular ions on the reactivation of human spermatozoa preserved in electrolyte-free solution. AB - It has previously been reported that human spermatozoa preserved in an electrolyte-free solution can survive for several weeks at 4 degrees C. However, the motility of spermatozoa after preservation cannot be restored when incubated at 37 degrees C, unless reactivated by extracellular alkalisation. Under weak acidic conditions, the reactivation is induced by > or = 10 mmol l-1 Na+ and inhibited by a Na(+)-H+ antiporter inhibitor. The addition of > or = 0.1 mmol l-1 K+ also induces the reactivation. In the present study, the reactivation was induced by > or = 0.1 mmol l-1 Rb+ or > or = 1 mmol l-1 Cs+ at an acidic pH. The maximum motility rate with K+, Rb+ or Cs+ was obtained at 10-20 mmol l-1 and inhibited by 10(-5)-10(-2) mol l-1 ouabain in a dose-dependent manner, while ouabain had no effect on the Na(+)-induced reactivation. The addition of K+ further increased sperm motility reactivated by Na+, which was also inhibited by ouabain. The addition of Ca2+ did not induce the reactivation or increase sperm motility reactivated by Na+ or K+. It was concluded that activation of the ouabain-sensitive Na(+)-K(+)-ATPase and Na(+)-H+ exchange mechanism has an important role in sperm motility. PMID- 10470412 TI - Inhibition of human sperm-zona-free hamster oocyte binding and penetration by protein C inhibitor. AB - Protein C inhibitor is a heparin-dependent serine protease inhibitor present in plasma at about 0.08 mumol l-1. Protein C inhibitor inhibits activated protein C and other coagulation factors. Previously, we described the presence of high protein C inhibitor levels in human semen (3.1 mumol l-1) and showed potential roles of the inhibitor in human reproduction. Here, we show that protein C inhibitor is present in an active form in follicular fluid at about 0.1 mumol l-1 and that purified, functionally active human plasma-derived and inactive, semen derived protein C inhibitor and a synthetic peptide derived from its sequence inhibited both binding and penetration of zona-free hamster oocytes by human sperm. The binding inhibition by protein C inhibitor was dose dependent, with 50% inhibition at 0.037 mumol l-1 inhibitor (45 +/- 17 sperm per egg versus 90 +/- 23 in control experiments). The inhibitor also blocked in a dose-dependent manner the penetration of zona-free hamster eggs by human sperm (20 +/- 7% fertilized eggs at 0.1 mumol l-1 protein C inhibitor versus 55 +/- 10% in control experiments). Polyclonal antiprotein C inhibitor or antipeptide antibodies partially abolished the effect of protein C inhibitor and peptide on the inhibition of the binding and penetration of zona-free hamster oocytes by human sperm. The effect of the protein C inhibitor was not dependent on its antiprotease activity since purified semen-derived protein C inhibitor which did not have antiprotease activity gave comparable results. We conclude that protein C inhibitor may be involved in human reproduction at several steps, including the fertilization process. PMID- 10470413 TI - Morphometric studies on the testes of rats treated neonatally with oestrogen and subsequently with gonadotrophins and testosterone. AB - The experiments involved male rats, which were given a single subcutaneous dose of 1 mg stilboestrol on the first day of life. Beginning on day 28, subgroups of the rats received either gonadotrophins or testosterone for 39 days. The weight of the testes, serum luteinizing hormone and testosterone levels were determined while sections of the testes were subjected to morphological analysis and morphometric measurements, based on computerized techniques. The results demonstrated that a single dose of oestrogen caused a reduction in the cross sectional area of the seminiferous tubules and a reduction in the thickness of the seminiferous epithelium, accompanied by inhibition of spermatogenesis. The number of and area occupied by Leydig cells, as well as the size of their cell nuclei, were also diminished, and the levels of serum testosterone decreased by 73%. All the experimental animals manifested significantly increased serum luteinizing hormone levels. Stimulation with gonadotrophins markedly increased the number of Leydig cells, their size and the size of their cell nuclei. This was associated with significantly increased levels of serum testosterone. Under these conditions, the cross-sectional area of the seminiferous tubules and the thickness of seminiferous epithelium remained less than those in the untreated controls. Following stimulation with testosterone the pattern of the seminiferous tubules resembled that noted after stimulation with gonadotrophins; the number of Leydig cells was markedly reduced but the size of both the cells themselves and of their nuclei approached normal values. PMID- 10470414 TI - Human sperm penetration in different media. AB - Sperm penetration into bovine cervical mucus and hen egg white using capillary tube penetration was investigated to verify the suitability of the capillary tube penetration test with hen egg white as a test of human sperm function. Semen samples from 50 consecutive patients were used for penetration tests and spermatozoa of a further 10 semen samples were penetrated into bovine cervical mucus and hen egg white for special motility assessment by computer-assisted motility analysis. Penetration tests revealed the well-known different ability of spermatozoa to penetrate into cervical mucus and a different penetration of spermatozoa into egg white for two nearly equal groups (n = 24 and n = 26, respectively). One group showed penetration comparable with cervical mucus and one group a very fast penetration up to the limit of the scale of measurement. Motility assessment of spermatozoa that penetrated into cervical mucus and egg white revealed significant differences in straight-line velocity, linearity and lateral head displacement. The number of spermatozoa selected actively during the penetration procedure was significantly higher in cervical mucus than in hen egg white. Spermatozoa selected by bovine cervical mucus and hen egg white exhibited a different motility pattern. There was significantly better linearity and less lateral head displacement in egg white than in cervical mucus. Sperm penetration into hen egg white appeared to be influenced by different sources of egg white. PMID- 10470415 TI - NIH orders Duke Medical Center to stop studies. PMID- 10470416 TI - When bombs put a stop to cancer treatments. PMID- 10470417 TI - NIH fails to respect deadline for prostate cancer plan. PMID- 10470418 TI - The challenge of response shift for quality-of-life-based clinical oncology research. PMID- 10470419 TI - Microsatellite instability (MSI) in non-colonic, non-HNPCC tumors: 'instable' evidence? PMID- 10470420 TI - Chemotherapy for brain metastases of lung cancer: a review. AB - In lung cancer patients brain metastases develop with a high frequency. For years radiotherapy has been the standard treatment for these patients. Here we review the experience with chemotherapy for brain metastases in lung cancer patients. The concept of the brain as pharmacological sanctuary site when brain metastases are present is challenged and it is argued that chemotherapy does play a role in this situation. Recent clinical trials indicate that the combination of chemotherapy and radiotherapy may become the standard treatment for lung cancer patients with brain metastases. It is unclear whether for micrometastatic disease to the brain, blood brain barrier function is of importance for the outcome of chemotherapy in lung cancer patients with respect to the development of overt brain metastases. Areas of improvement of delivery of cytotoxic agents to the brain when brain metastases have not yet developed are discussed. PMID- 10470421 TI - Using evaluations of diagnostic tests: understanding their limitations and making the most of available evidence. PMID- 10470422 TI - START: a European state-of-the-art on-line instrument for clinical oncologists. AB - START ('State-of-the-Art Oncology in Europe'), a freely available resource on the Internet, is a European 'information base' of current clinical approaches to human tumours. Its aim is to help clinical oncologists make appropriate clinical decisions by providing them with updated information reflecting state-of-the-art cancer treatment as perceived by the European oncology community. It is based upon contributions from authors and internal reviewers from all over Europe as selected by START's editorial board under the supervision of an advisory board and a scientific committee. Close collaborations with the main European cancer societies are ongoing. An external feedback process augments the mechanisms for rendering START a truly European instrument. START is concerned with evidence based cancer medicine, and the main clinical options are thus codified and their bases indicated in accord with a scale worked out from the perspective of clinical decision-making. Therapeutic options may be 'standard', 'investigational', or 'suitable for individual clinical use' (within the context of a decision made jointly by the patient and the physician). The goal of instruments such as START is to improve the quality of patient care. In addition, START hopes to make contributions to the methodology by which medical research is transformed into clinical decisions. PMID- 10470423 TI - Quality of life as subjective experience: reframing of perception in patients with colon cancer undergoing radical resection with or without adjuvant chemotherapy. Swiss Group for Clinical Cancer Research (SAKK) AB - PURPOSE AND BACKGROUND: We examined whether patients with colon cancer undergoing surgery with or without adjuvant chemotherapy change the internal standards on which they base their quality-of-life (QL) estimation, and, if they do so, whether this reframing alters interpretation of QL findings. These questions were addressed within a randomized clinical trial of the Swiss Group for Clinical Cancer Research (SAKK 40/93). PATIENTS AND METHODS: After radical resection of adenocarcinoma of the colon (pT1-4pN > 0M0 and pT3-4pN0M0) and perioperative chemotherapy, patients were randomized to three treatment arms: observation only (A), 5-FU 450 mg/m2 plus Levamisol (B), or 5-FU 600 mg/m2 (C). QL was measured by linear analogue self-assessment indicators. Patients estimated their pre-surgery QL both before surgery and retrospectively thereafter, and their pre-adjuvant QL both at the beginning of randomly assigned chemotherapy or observation and retrospectively about two months later. Thereafter, current QL was assessed. Paired t-tests were used to test the hypotheses of no change. RESULTS: Overall, 187 patients with at least one pair of corresponding questionnaires were analyzed. Patients estimated their pre-surgery QL after surgery significantly lower than before and their pre-adjuvant QL under treatment or observation also lower than at the beginning. In the adjuvant phase, in contradiction to our hypothesis, chemotherapy had almost no impact on these changes attributed to reframing. Conventionally assessed changes indicated an improvement in QL. Patients with treatment C reported less improvement in functional performance than those with B or those under observation (P = 0.04). Patients with treatment B indicated a greater worsening in nausea/vomiting than those with C, whereas patients with observation only showed an improvement (P = 0.0009). After adjustment of current QL scores under treatment or observation to patients' retrospective estimation, the treatment effects were diluted but the overall improvement was substantially amplified in most QL indicators. CONCLUSIONS: Patients with colon cancer substantially reframe their perception in estimating QL both under radical resection and under adjuvant chemotherapy or observation. This effect is an integral part of patients' adaptation to disease and treatment. An understanding of this phenomenon is of particular relevance for patient care. Its role in evaluating QL endpoints in clinical trials needs further investigation. PMID- 10470424 TI - Microsatellite instability in gastric MALT lymphomas and other associated neoplasms. AB - BACKGROUND: Microsatellite instability (MSI), caused by a reduced efficacy of the DNA mismatch repair (MMR) machinery, represents a type of genomic instability frequently detected in HNPCC spectrum cancers and in a subset of sporadic carcinomas. The involvement of MSI in the pathogenesis of gastric lymphoma of mucosa-associated lymphoid tissue (MALT) has never been conclusively investigated. In this study, we tested the presence of MSI in tumor samples of patients harboring both MALT lymphomas and other types of malignancies. MATERIALS AND METHODS: We examined 10 microsatellite loci (D3S11, D3S1261, D3S1265, D6S262, D6S193, BAT-26, BAT-25, D17S250, APC, D2S123) out of a total of 34 primary tumors from 14 patients with MALT lymphomas and one or more additional neoplasms. The patients' MSI results were also tested for an association with a positive family history of cancer. RESULTS: MSI, defined by the presence of microsatellite alterations in more than 40% of the examined loci, was scored negative in all tumors studied, and pedigree analysis failed to identify any condition of familial cancer among the patients examined. CONCLUSIONS: The present study suggests that defects in DNA mismatch repair do not contribute significantly to the molecular pathogenesis of MALT lymphomas and associated neoplasms. PMID- 10470425 TI - Is adjuvant tamoxifen used optimally in the treatment of breast cancer? Results of an Italian survey. AB - BACKGROUND: Institutional and physician-related factors can influence the way in which physicians interpret research results. The aim of this study was to determine what physicians know about, and their opinions of, hormone treatment in breast cancer patients, and the factors comprising their medical decision-making. MATERIALS AND METHODS: A questionnaire was mailed to a random sample of physicians inquiring as to their preferences with respect to adjuvant tamoxifen, and the usual duration of the treatment applied in various clinical scenarios (according to a woman's menopausal status, the oestrogen receptor status and the stage of disease). RESULTS: Of 500 physicians identified, 38% returned the questionnaire. Of the non-responders, a random sample of 60 physicians was interviewed by phone. The total number of available questionnaires was 250 (50%). About 3/4 of the doctors would prescribe tamoxifen in older ER+ women and 30%-40% in post-menopausal ER-patients, but only 2/5 would do so in younger ER+ women. The vast majority of physicians considered five years as standard for ER+ patients. Nevertheless, about 1/4 of the doctors chose a shorter treatment duration for node-negative, pre-menopausal patients. A minority of physicians used tamoxifen for longer than five years. Older clinicians were less likely to prescribe tamoxifen, particularly for low-risk patients. CONCLUSIONS: According to the data of the recent EBCTG overview, an additional 20,000 lives could be saved worldwide each year if tamoxifen were given to all early breast cancer patients with hormone-sensitive disease, irrespective of age and disease stage, and for a minimum of five years. Our study, involving a representative sample of physicians practicing in Italy, shows that tamoxifen is not used optimally, with a substantial under-use in younger women and women with node-negative disease. PMID- 10470426 TI - High-dose epirubicin and cyclophosphamide every two weeks as first-line chemotherapy for relapsing metastatic breast cancer patients. AB - BACKGROUND: Metastatic breast cancer remains incurable with conventional chemotherapy. For any specific chemotherapy, higher dose intensity may be achieved with either increased doses per cycle, or shortened intervals between courses, or both. We demonstrate here the feasibility and encouraging results of a high-dose combination regimen administered every two weeks. PATIENTS AND METHODS: Women with metastatic breast cancer were treated every 14 days for 6 courses with 75 mg/m2 epirubicin and 1200 mg/m2 cyclophosphamide, followed by conventionally-delivered (q 3-4 weeks) chemotherapy. The treatment was to be resumed regardless of the neutrophil count, except in instances of febrile neutropenia. Prophylactic oral antibiotherapy was given, while hematopoietic growth factors and stem cell support were not employed. RESULTS: Eighty-six patients were treated between May 1986 and June 1995. Their median age was 43 years (26-69). Grade 3-4 neutrophil toxicity was observed after 86% of the courses, resulting in febrile neutropenia in 5%-18% of the patients, and the rehospitalization of 5%-10%. The median given/planned dose intensity was 97% (79 106). The objective response rate in 84 evaluable patients was 54% (95% confidence interval (95% CI): 43-65), with a complete response rate of 11%, and a 14% rate of outright progression. Median progression-free survival was 16 months and median overall survival 32 months. Multivariate analysis retained previous adjuvant chemotherapy as a negative survival prognostic factor. CONCLUSIONS: This dose-intensive anthracycline-based regimen is feasible with manageable morbidity despite pronounced myelotoxicity, and yields encouraging survival rates. PMID- 10470427 TI - Concurrent carboplatin/5-fluorouracil and radiotherapy for recurrent cervical carcinoma. AB - BACKGROUND: Results of salvage therapy in patients with carcinoma of the uterine cervix recurrent after surgery have been dismal even when the disease was apparently confined to the pelvis. Concurrent chemoradiation is one of the several avenues being investigated to improve these results. PATIENTS AND METHODS: Thirty-five women with recurrent cervical carcinoma were enrolled in the trial. Twenty-eight patients (80%) had disease limited to the central pelvis (ten), lateral pelvis (fourteen) and vagina (four) and seven had paraortic metastases. Patients were treated with a combination of external radiotherapy (50 70 Gy) along with three cycles of 5-fluorouracil (1000 mg/m2/24-hour continuous infusion days 1-4) and carboplatin (75 mg/m2 in bolus days 1-4). RESULTS: Treatment was well tolerated, with 30 patients (86%) completing the protocol as planned. Acute toxicity was severe but manageable; 11 patients (31%) experienced grades 3-4 acute toxicity. Late morbidity occurred in five patients (14%). Overall response rate was 74% (11 partial responses and 15 complete). After a median follow-up of 27 months (range 18-90), 13 patients (37%) are alive without disease, 4 (11%) are alive with persistent disease and 18 (52%) are dead of their disease. Actuarial two-year survival rate for the series as a whole is 44% and three-year survival is 25%. Stage of the primary disease, site of recurrence, interval from the primary therapy to recurrence, lymph node involvement, ureterohydronephrosis at the time of recurrence and diameter of the relapse are the most significant factors for survival, while complete response is related to diameter and site of relapse and lymph node status at the time of relapse. CONCLUSION: The acceptable toxicity, high response rate and satisfying survival would suggest that concomitant carboplatin/5-fluorouracil and radiotherapy is a safe and tolerable treatment for recurrent cervical carcinoma. Further studies are needed to demonstrate an eventual survival benefit of this type of chemoradiation over standard radiotherapy alone and to identify the subsets of patients who in particular might benefit from this. PMID- 10470428 TI - Small-cell lung cancer: patients included in clinical trials are not representative of the patient population as a whole. AB - PURPOSE: To determine the clinical characteristics of the subpopulation of patients not included in clinical trials, their outcome, and the reasons for their ineligibility and non-participation. PATIENTS AND METHODS: We studied 57 patients (out of 178 consecutive patients with SCLC), who were not included in any of the three successive clinical trials completed at our center during the study period. We also compared 37 patients excluded from the largest clinical trial to their 73 included counterparts. RESULTS: Reasons for ineligibility (n = 53) included low Karnofsky index (n = 17), advanced age (n = 12), non-feasible long-term follow-up (n = 12), previous history of cancer (n = 8), contraindication for anthracyclines (n = 5), and other medical reasons (n = 11). Only four eligible patients were not included in the trials. As compared to patients included in the studies, non-included patients had a significantly lower Karnofsky index, were older, presented more frequently with metastatic disease, and had a lower response rate to treatment and a shorter survival. However, exclusion from the trial was not an independent prognostic factor by multivariate analysis. CONCLUSIONS: Selection biases were unlikely in the three trials, based on the high ratio of included/eligible patients. However, the subgroup of patients included in the trials was not representative of the patient population as a whole because of restrictive eligibility criteria. Results from published clinical trials to the overall population should be extrapolated only with caution. We suggest that the proportion and major characteristics of ineligible and non-participating patients be mentioned in any publication of a clinical trial. PMID- 10470429 TI - Possible improved survival of patients with stage IV AJCC melanoma receiving SRL 172 immunotherapy: correlation with induction of increased levels of intracellular interleukin-2 in peripheral blood lymphocytes. AB - PURPOSE: This phase I-II study was designed to assess safety and clinical efficacy of SRL 172 vaccine in patients with advanced stage IV (AJCC) malignant melanoma. Induction of intracellular cytokines (IL-2 and INF-gamma) in peripheral blood lymphocytes (PBLCs) from these patients was assayed and correlated to clinical outcome. PATIENTS AND METHODS: SRL 172 was administered intradermally to 24 patients with stage IV malignant melanoma, initially at 15-day intervals for three vaccinations and then at monthly intervals. Lymphocyte activation for cytokines in PBLCs was assayed prior to each vaccine administration using a FACS based intracellular cytokine assay. Survival was compared to historical controls. RESULTS: The vaccination schedule resulted in sustained intracellular IL-2 induction in PBLCs in 9 of 23 patients (39%) who received at least three doses. Cytokine induction became apparent within the first three administrations of vaccine and was maximal at 8-12 weeks. Induction of intracellular IL-2 production (group 1) was associated with improved survival (P < 0.036). The median survival of the nine patients demonstrating IL-2 induction was 59 (95% confidence interval (95% CI): 47-71) weeks compared to 31 (95% CI: 18-44) weeks for the non-inducers. Induction of INF-gamma (group 2) was found in 10 patients and 6 patients had IL-2 and INF-gamma induction (group 3). There was no survival advantage for these patient groups. Although no objective responses were documented the group as a whole had a median survival of 44 (95% CI: 31-59) weeks which is better than that of historical controls. SRL 172 was safe and well tolerated. CONCLUSION: SRL 172 is effective in inducing intracellular IL-2 responses in PBLCs of a significant number of patients with stage IV (AJCC) melanoma. This is correlated with improved survival. The survival analysis is sufficiently encouraging to suggest that further prospective trials are justified. The methodology we present in this study may help in developing surrogate markers that will allow rational immunotherapeutic strategies to be designed for cancer patients. PMID- 10470430 TI - Application of the International Prognostic Scoring System for myelodysplastic syndromes. AB - BACKGROUND: In March 1997 an international workshop introduced a new International Prognostic Scoring System (IPSS) for MDS. The goal of the present study was to apply the IPSS to a large group of MDS patients from one centre and to compare it to the FAB-classification. PATIENTS: One hundred eighty-four MDS patients were included on the basis of similar criteria as used by the workshop but some of them (30) received AML-type therapy. RESULTS: The IPSS separated our patients into distinctive prognostic subgroups (P = 0.0001). Median survival was respectively 6.5, 2.6, 1.3 and 0.75 years for the low-risk (22% of patients), the intermediate-1-risk (INT-1) (46%), the intermediate-2-risk (INT-2) (25%) and the high-risk group (7%). The IPSS also discriminated within each of the FAB categories: RA patients (58 patients) were present in low-risk, INT-1-risk and INT-2-risk subgroups, RARS patients (23) were separated into low-risk and INT-1 risk subgroups. RAEB patients (53) were distributed predominantly between INT-1 risk and INT-2-risk groups, RAEB-t patients (23) between INT-2-risk and high-risk subgroups. CMML patients (27) were present in the low-risk, the INT-1-risk and the INT-2-risk group. CONCLUSIONS: Our results confirm the effectiveness of the IPSS in predicting clinical outcome in MDS patients and indicate that it is an improved method compared to the FAB-classification. PMID- 10470431 TI - Sequential administration of temozolomide and fotemustine: depletion of O6-alkyl guanine-DNA transferase in blood lymphocytes and in tumours. AB - BACKGROUND: The DNA repair protein O6-alkylguanine-DNA alkyl transferase (AT) mediates resistance to chloroethylnitrosoureas. Agents depleting AT such as DTIC and its new analogue temozolomide (TMZ) can reverse resistance to chloroethylnitrosoureas. We report the results of a dose finding study of TMZ in association with fotemustine. PATIENTS AND METHODS: Twenty-four patients with metastatic melanoma or recurrent glioma were treated with escalating dose of oral or intravenous TMZ ranging from 300 to 700 mg/m2, divided over two days. Fotemustine 100 mg/m2 was given intravenously on day 2, 4 hours after TMZ. AT depletion was measured in peripheral blood mononuclear cells (PBMCs) and in selected cases in melanoma metastases and was compared to TMZ pharmacokinetics. RESULTS: The maximum tolerated dose (MTD) of TMZ was 400 mg/m2 (200 mg/m2/d) when associated with fotemustine the 2nd day with myelosuppression as dose limiting toxicity. The decrease of AT level in PBMCs was progressive and reached 34% of pretreatment values on day 2. There was however wide interindividual variability. AT reduction was neither dose nor route dependent and did not appear to be related to TMZ systemic exposure (AUC). In the same patients, AT depletion in tumour did not correlate with the decrease of AT observed in PBMCs. CONCLUSIONS: PBMCs may not be used as a surrogate of tumour for AT depletion. Further study should concentrate on the pharmacokinetic pharmacodynamic relationship in tumour to provide the basis for individually tailored therapy. PMID- 10470432 TI - A comprehensive assessment of satisfaction with care: preliminary psychometric analysis in an oncology institute in Italy. AB - BACKGROUND: Little is known about patients' perception of the quality of the care they receive in oncology hospitals. We developed a 61-item comprehensive assessment of satisfaction with care (CASC) to evaluate the competence of hospital physicians and nurses, as well as aspects of care organisation and hospital environment. The aims of this study were to define the structure of the CASC and assess the internal consistency and convergent and discriminant validity of its scales. PATIENTS AND METHODS: Three hundred ninety-five consecutive cancer patients discharged from an oncology institute in Italy were asked to complete the CASC at home and return it in a self-addressed envelope. RESULTS: Two percent of the patients refused to participate and 25% failed to return the questionnaire. Separate factor analyses of the CASC sub-scales disclosed the perceived extent of doctors' and nurses' availability, coordination, human quality, technical competence, provision of psychosocial care and information, as well as the patients' general satisfaction, perception of the organisation of their care, access and comfort. Multi-trait scaling analysis was carried out on item-grouping resulting from factor analyses. High levels of internal consistency and convergent validity were obtained but discriminant validity could be improved. CONCLUSIONS: Results of present psychometric testing of the CASC forecast adequate properties. This will be confirmed by repeating these analyses in a cross-cultural setting. PMID- 10470433 TI - The management of non-small-cell lung cancer: a case history. AB - Accurate assessment and treatment of the patient with lung cancer requires a team approach involving respiratory physicians, cardiothoracic surgeons, oncologists and the palliative care team. Adequate staging and assessment of prognostic factors are essential before deciding what treatment is appropriate for an individual patient. Surgery is the mainstay of treatment for early disease. Patients with medically inoperable stage 1 (T1, T2, N0) tumours should be considered for radical radiotherapy; additional chemotherapy in early stage disease may offer an additional survival advantage, but its overall role can only be assessed by further clinical trials. In more locally advanced tumours radical radiotherapy has never been formally tested. It is however, often used in patients where the tumour can be encompassed safely within a radiation field. This will depend on total dose and fractionation schedule as well as the volume of tissue irradiated. Neo-adjuvant chemotherapy prolongs survival in these patients. As only a few patients are cured, symptom control and quality of life are usually the most important goals of management and can be achieved by a variety of interventions. It is disappointing that in such a common disease less than 5% of patients are entered into clinical trials. Without such evidence the therapeutic outcomes in NSCLC cannot be improved. PMID- 10470434 TI - Phase II study of gemcitabine in ovarian cancer. AB - This phase II study evaluated the response rate and toxicity of single-agent gemcitabine in 40 women with epithelial ovarian cancer, previously treated with platinum-based chemotherapy. Patients had stage III or IV disease and progressive disease 1-12 months after the last treatment. Gemcitabine 1250 mg/m2 was administered on days 1, 8 and 15 of each 28-day cycle as a 30-minute infusion. The overall response rate to gemcitabine was 22% (95% confidence intervals: 10 39%). Responses to gemcitabine were observed in patients with platinum-refractory disease, which suggests no cross resistance to platinum. Gemcitabine was well tolerated and no grade 4 toxicity was seen. This study confirms that gemcitabine is active and well tolerated in pre-treated women with ovarian cancer. PMID- 10470435 TI - Thalidomide in patients with cachexia due to terminal cancer: preliminary report. PMID- 10470436 TI - A phase I study of sequential vinorelbine followed by paclitaxel. AB - BACKGROUND: In vitro experiments suggest that administration of vinorelbine preceding paclitaxel results in synergistic cytotoxic effects. A phase I dose escalation trial of vinorelbine daily x 3 with paclitaxel on day 3 repeated every 28 days in metastatic breast cancer patients was completed. PATIENTS AND METHODS: Female patients, PS 0-2, without evidence of CNS disease or prior neuropathies were treated with vinorelbine at dose levels 7, 10, 13 mg/m2 per day and paclitaxel over three hours at dose levels of 135, 175, and 200 mg/m2. RESULTS: Twenty-eight patients with six dose levels were studied. At dose level 1, patients developed intolerable but reversible neutropenia. Subsequent dose levels required filgrastim. Dose limiting toxicities were myalgia and fatigue at vinorelbine 13 mg/m2/day and paclitaxel 200 mg/m2. Neuropathy was minor. Twelve of twenty-five patients with measurable disease had a rapid response which did not correlate with dose level. CONCLUSIONS: Sequential administration of these two agents demonstrates activity in breast cancer patients. Phase II dosing on this schedule should be vinorelbine 13 mg/m2/day x 3 and paclitaxel 175 mg/m2. With proper selection of patients, concern about neurologic toxicity should not impede future trials of vinorelbine with paclitaxel. PMID- 10470437 TI - Does pain at tumor site during vinorelbine infusion affect treatment of recurrent head and neck cancer patients? PMID- 10470438 TI - Involvement of Fas/FasL system in the pathogenesis of autoimmune diseases and Wilson's disease. AB - The interaction of Fas with FasL has been demonstrated to be implicated in the pathogenesis of several autoimmune and liver diseases. Recently, attention has been focused on the hypothesis that thyrocytes and beta cells undergo massive Fas/FasL-mediated apoptosis during autoimmune response. Similarly, hepatocyte cell death occurring following copper accumulation points towards the same mechanism. PMID- 10470439 TI - Virokines: mediators of virus-host interaction and future immunomodulators in medicine. AB - A decade ago, after the discovery of the major secretory protein of vaccinia virus with structural similarity to complement control, the term virokine was coined. The term virokine, simply refers to a virally encoded secretory protein. During the past decade several virokines were discovered and most of them have been found to have immunomodulatory effects. A subset of virokines which resemble cytokine/chemokine receptors have been termed viroceptors. Examples of viroceptors include the TNF receptor homolog and the IL-1 beta receptor homolog. During the past several years animal models have been developed to try to understand the in vivo role of the virokines. It has become evident from at least two studies that quite a few of the virokines down-regulate the inflammatory response elicited during infection. This down-regulation at least in one model system seems to indicate that it ensures the preservation of the viral habitat (site of infection). One obvious spinoff of the research on virokines is a new class of immunomodulators has become available as therapeutics in alleviating the symptoms of inflammatory disease conditions. PMID- 10470440 TI - How does the major histocompatibility complex influence behavior? AB - The major histocompatibility complex (MHC) encodes highly polymorphic cell surface glycoproteins that form the basis of immunological individuality. It has been postulated that MHC besides its role in immune defence also determines behavior through formation of specific odor cues. In addition several studies have implicated MHC disassortative odor and mating preferences in mice, rats, and humans. Further leading studies have suggested that mice and humans prefer to mate with MHC dissimilar individuals. Although the latter remains controversial, MHC dependent mating preferences provide a potentially important selective factor driving the polymorphisms of the MHC genes. Here we review some of the available data and hypotheses on the influence of the MHC on behavior and discuss its molecular and chemical basis. PMID- 10470441 TI - Cyclolinopeptides and their analogs--a new family of peptide immunosuppressants affecting the calcineurin system. AB - The results of the investigation of immunosuppressive activity of cyclolinopeptide A (CLA--cyclic hydrophobic nonapeptide present in the linseeds) and its analogs are discussed. The results obtained for other natural cyclic peptides showing structural similarities with CLA (antamanide, cycloamanides, hymenistatin, hymenamides) are also reviewed. It results from these investigations that the molecular mechanism of the CLA action is the same as that of cyclosporin A and FK-506 compound, i.e. it consists in formation of the complex with cyclophilin and inhibition--in this form--of the phosphatase activity of calcineurin. The results also suggest that the immunosuppressive activity of these compounds resides in their--Pro-Xxx-Phe- fragment, where Xxx is a hydrophobic (e.g. Leu, Val) or aromatic amino acid residue. PMID- 10470442 TI - Nitric oxide, heparin and procaine treatment in experimental ceruleine-induced acute pancreatitis in rats. AB - The aim of the study was to investigate the impact of L-arginine (nitric oxide donor), L-NNA (NO synthase inhibitor), heparin and procaine on the pancreas' microcirculation, serum interleukin 6 (IL-6) level, and microscopic alterations of the pancreatic gland in acute pancreatitis (AP) in rats. AP was induced by 4 i.p. injections of cerulein (15 micrograms/kg/h). Microcirculatory values of the pancreas were measured by means of laser Doppler flowmetry 5 h after the first cerulein injection. Remarkable morphologic changes in the pancreas, including parenchymal necrosis, an elevation of serum IL-6 activity, and significant drop of pancreatic capillary perfusion was observed in rats with NO synthase inhibition. L-arginine improved the pancreatic microcirculation but worsened the microscopic alterations within the pancreas. Heparin had a beneficial effect on the microcirculatory values, serum IL-6 activity, and morphologic changes. Procaine had no effect on the course of AP. Authors conclude that heparin, improving the pancreatic capillary blood perfusion, may be considered as a promising therapeutic agent in acute pancreatitis. PMID- 10470443 TI - Tumor infiltrating lymphocytes in HLA+ and HLA- laryngeal cancer--quantitative approach. AB - In search of factors governing the accumulation of tumor infiltrating lymphocytes (TIL), frozen sections from fresh surgical specimens of laryngeal carcinoma (n = 36) were tested by alkaline phosphatase-anti-alkaline phosphatase (APAAP) immunohistochemistry for monomorphic determinants of HLA class I and class II expression on tumor cells and for the distribution of lymphoid cells bearing CD differentiation antigens. Cell subsets were quantitated in two tumor compartments, tumor mass and tumor stroma, by computer-assisted image analysis. In a portion of examined samples lymphoid cell suspension was isolated from cancerous tissues and assessed by flow cytometry. It has been found that T cells, localized mostly in tumor stroma, were predominant cell population in the tumor microenvironment. Their ability to penetrate tumor mass but not tumor stroma, by CD8+ T cells in particular, but also by natural killer (NK) cells, was associated with HLA class I antigen expression on tumor cells. In flow cytometric analysis activated T lymphocytes (CD3+DR+) were abundant in HLA+ tumors as compared to HLA ones. In 4 year follow up of 20 patients the mortality was higher in HLA- group but the data were not statistically significant. These results show that HLA class I expression on tumor cells favor penetration of cytotoxic lymphoid cells into tumor mass, at least in the laryngeal cancer. PMID- 10470444 TI - Adhesion molecule expression stimulated by Bacteroides thetaiotaomicron cell surface antigens. AB - Bacteroides thetaiotaomicron, a Gram-negative anaerobic rod belonging to the Bacteroides fragilis group (BFG), is involved in many systemic and local, most frequently suppurative infections in man. The cell envelope of these rods is composed of two carbohydrate-containing antigens: lipopolysaccharide (LPS) and capsular polysaccharide (CPS). Adhesion molecules ICAM-1, VCAM-1 and E-selectin (ELAM-1) are induced on the endothelial cells by mediators of inflammation. The aim of this study was to assay the ability of B. thetaiotaomicron surface antigens to induce adhesion molecule expression on the endothelial cells. The influence of LPS and CPS on the expression of adhesion molecules on HMEC-1 cell line was examined in an ELISA test. ELISA was performed with monoclonal mouse anti-human: ICAM-1, VCAM-1 and E-selectin antibodies of the IgG class. B. thetaiotaomicron lipopolysaccharides revealed the ability to induce ICAM-1, VCAM 1 and E-selectin expression on the endothelial cells. Their activities were similar, but lower than the activity of Eschericha coli LPS. ICAM-1 was the most stimulated adhesion molecule. The strongest activation by LPS was achieved at the concentrations of 10.0 and 1.0 micrograms/ml. The ability of capsular polysaccharide to induce the expression of adhesion molecules was considerably weaker. PMID- 10470445 TI - Isolation of human lutropin from woman urine and comparison of its properties with pituitary hormone. AB - Lutropin was isolated from woman preovulatory urine by immunoaffinity chromatography using a column with monoclonal antibody (mAb) anti-beta hLH(37) coupled to Sepharose CL 4B. As it was demonstrated the isolated hormone, as well as pituitary lutropin, were separated in SDS-PAGE into several well visible fractions with 30-94 kDa molecular mass and scarcely visible fractions with 14-20 kDa. All fractions reacted only with mAb anti-alpha hCG(99)-HRP but not with mAb anti-beta hLH-HRP. Pretreatment of pituitary lutropin with PN-Gase F did not affect its electrophoretic pattern. After boiling the hormone with SDS and beta ME no fractions in SDS-PAGE were observed. No substantial differences in affinity chromatography on Con A-Sepharose between pituitary and urinary lutropin were noted. Some differences between these two hormone preparations were observed in assays performed with several ELISA variants. Using two pairs of mAbs anti-beta hLH for ELISA technique no hormone was assayed in urine samples from women, collected between 12 and 16 days of menstrual cycles. PMID- 10470446 TI - Antifungal activity of 2-(4-chlorophenyl)-1,2-benzisoselenazol-3(2H)-one, the analog of Ebselen. AB - The antimicrobial activity of 8 selenoorganic compounds (3-benzisoselenazoles, 1 benzisoselenazolone oxide and 4-disaryl diselenides) was investigated. It was found that selenoorganic compounds from benzisoselenazolone group suppress growth of some fungi and bacteria. The growth of Saccharomyces cerevisiae sigma 127-8b and Candida albicans 258 strains was strongly inhibited by the 2-(4-chlorophenyl) 1,2-benzisoselenazol-3(2H)-one. Also Ebselen and 2-acetyl-1,2-benzisoselenazol 3(2H)-one caused strong inhibition of Saccharomyces cerevisiae sigma 127-8b growth but a lower effect was observed in assays with Candida sp. strains. Benzisoselenazolones were also found to have antibacterial activities. They significantly reduced the growth of Gram-negative Escherichia coli K-12 ROW and Gram-positive Staphylococcus aureus 209P (Oxford) bacteria strains. PMID- 10470447 TI - Investigation of influence of Proteus mirabilis LPS polysaccharide part on the murine immune cells activation. AB - The biological activities were investigated of Proteus mirabilis lipopolysaccharides (LPSs) and their fragments, namely, the induction of nitric oxide (NO) and interleukin 1 (IL-1) synthesis by murine macrophages and the proliferation of murine spleen cells. The O-specific polysaccharide (F1), core oligosaccharide (F2) and lipid A were as effective as intact LPS in stimulating murine macrophages to produce NO. IL-1 synthesis was also induced by all studied types of endotoxins (S, Ra, Re) and partial structures, however F1, F2 and lipid A fractions required the presence of serum. In contrast to LPS, the O-specific polysaccharide, core oligosaccharide and lipid A were not able to induce the blast response of murine non-adherent splenocytes. PMID- 10470448 TI - Gaining weight: the scientific basis of increasing skeletal muscle mass. AB - Most athletes today tend to have a larger muscle mass than their predecessors. Better training and nutrition practices are responsible for much of this difference, but whatever the mechanism, the balance between muscle protein synthesis and breakdown must be in favor of increased muscle protein. Applying new techniques for measuring whole body and muscle protein synthesis to resistance exercise has led to some interesting results. In the recovery period following resistance exercise, both muscle protein synthesis and breakdown are accelerated in the fasted state. Ingestion of carbohydrate or carbohydrate and protein during recovery further increases muscle protein synthesis, due in part to an improved anabolic hormone environment. In addition, the anabolic effect of a resistance training bout may last well beyond 48 hours. Using information obtained from research studies, better training and dietary practices can optimize the benefits from resistance training. PMID- 10470449 TI - Leptin and reproduction: is it a critical link between adipose tissue, nutrition, and reproduction? AB - Exercise-associated reproductive disorders are frequently reported among recreationally active and elite female athletes. Although an association between exercise and menstrual disorders has been established, the mechanism by which exercise disrupts reproductive function remains unknown. Recent findings suggest that low energy availability rather than inadequate body fatness or exercise stress is likely the mechanism by which exercise impinges negatively on the hypothalamic-pituitary-ovarian axis in female athletes. The peripheral signal that informs the neural network of energy availability remains unknown. The identification of the adipocyte-derived ob gene product, leptin, and subsequent findings of its association with reproduction in both rodents and humans, led to speculations that it may be involved in the interactions between nutrition and reproduction. This review article focuses on leptin's role in modulating reproduction, and in particular, as a peripheral signal of nutritional status that integrates adipose tissue, nutrition, and reproduction in female athletes. PMID- 10470450 TI - Reproducibility of resting heart rate variability with short sampling periods. AB - The purpose of this study was to determine whether resting heart rate variability (HRV) is reproducible with short sampling measurement periods using an office based personal computer measurement system. Eight healthy active women participated in ECG analyses on 2 days within 1 week under controlled environmental and physiological conditions. After they rested for 10 minutes, a 10-min ECG was recorded. HRV was determined from a 2.5- and 5-min sample period using both time domain variables (meanRR and SDNN) and frequency domain variables (LF, HF, LF:HF). Repeated measures ANOVA found no significant differences between Day 1 and Day 2 for either sampling period (p > or = 0.23). For both the 2.5- and 5-min sampling periods, the intraclass correlations between days for the time domain variables showed good reproducibility (R = 0.86-0.90). The reproducibility of the frequency domain variable was only average (R = 0.67-0.96), with the LF:HF ratio yielding the higher R values. PMID- 10470451 TI - Effects of dehydration, hypohydration, and hyperhydration on tolerance during uncompensable heat stress. AB - The present study examined the effects of dehydration from prior exercise on subsequent exercise tolerance time (TT) that involved wearing nuclear, biological, and chemical (NBC) protective clothing. It was hypothesised that TT would be reduced in the dehydrated state. Ten men undertook continuous treadmill walking at 4.8 km.h-1 at 35 degrees C and 50% relative humidity, wearing NBC clothing while euhydrated (EU) or dehydrated (D) by 2.3% of body weight. Hydration status had no impact on thermoregulatory or cardiovascular responses during exercise. Also rectal temperature at exhaustion did not differ between EU (38.52 +/- 0.39 degrees C) and D (38.43 +/- 0.45 degrees C). Exercise TT during this uncompensable heat stress was reduced significantly for D (47.7 +/- 15.3 min) compared with EU (59.0 +/- 13.6 min). It was concluded that prior exercise leading to levels of dehydration to 2.3% of body weight, together with subsequent fluid restriction during exposure to uncompensable heat stress, impaired TT while wearing the NBC protective clothing. The integration of these findings together with other comparable studies that have examined the influence of hypo- and hyperhydration on TT while wearing NBC protective clothing revealed that hydration status has less effect on TT as the severity of uncompensable heat stress increases. PMID- 10470452 TI - Fetal responses to maternal strength conditioning exercises in late gestation. AB - Cardiovascular responses to strength conditioning exercises were examined in 12 healthy pregnant women and their unborn fetuses during the third trimester. A group of 12 healthy nonpregnant women of similar ages, parity, body height, and pre-pregnant body mass was also studied. Maternal heart rate and blood pressure and fetal heart rate (FHR) responses were measured in both the supine (30 degrees tilt) and seated postures during handgrip (HG), single-leg extension (SL), and double-leg extension (DL) exercise. Subjects performed 3 sets of 10 reps at 50, 70, and 90% of their 10-repetition maximum (10-RM) for each exercise in both postures. Pregnant subjects exhibited higher heart rates but similar blood pressure responses to control subjects under all experimental conditions. Significant increases were observed for the frequency of FHR accelerations (0.10 to 0.27/min) from rest to DL in the sitting posture at 90% RM. Moderate fetal bradycardia was observed occasionally in the tilted supine posture at rest and both during (SL, DL) and following (HG, SL, DL) exercise, suggesting that this posture should be avoided in late gestation. The results support the safety of moderate strength conditioning exercises in healthy pregnancy. PMID- 10470453 TI - Increasing exercise duration does not affect the postexercise elevation in esophageal temperature. AB - It has previously been observed that (a) following 15 min of intense exercise, esophageal temperature (Tes) remains elevated at a plateau value equal to that at which active vasodilation had occurred during exercise (i.e., esophageal temperature threshold for cutaneous vasodilation [ThVD]); and (b) exercise/recovery cycles of identical intensity and duration, when sequential, result in progressively higher Tes at the beginning and end of exercise. In the latter case, parallel increases in both the exercise ThVD and postexercise plateau of Tes were noted. This study was conducted to determine if the elevated postexercise Tes is related to increases in whole-body heat content. On separate occasions, 9 subjects completed 3 bouts of treadmill exercise at 70% VO2 max, 29 degrees C ambient temperature. Each exercise bout lasted either 15, 30, or 45 min and was followed by 60 min of inactive recovery. Esophageal temperatures were similar at the start of each exercise bout, but the rise in Tes during exercise nearly doubled from 1.0 degree C after 15 min of exercise to 1.9 degrees C after 45 min of exercise. There were no intercondition differences among the exercise ThVD (approximately 0.36 degree C above baseline) or postexercise plateau values for Tes (approximately 0.40 degree C above baseline). Thus the relationship between the ThVD during exercise and the postexercise Tes did not appear to be dependent on changes in whole-body heat content as produced by endogenous heating during exercise of different duration. PMID- 10470454 TI - Cancer staging. PMID- 10470455 TI - History and international developments in cancer staging. AB - OBJECTIVE: To inform the reader of the objectives of staging classification, and review history of the development of modern staging classifications in cancer. DESIGN: Review of the literature documenting the development of modern cancer staging systems with the emphasis on the history of the development of the TNM classification by the UICC and the history of the Canadian Committee on Cancer Staging. The underlying principles of the TNM system have been reviewed in the context of modern cancer practice. CONCLUSION: In the era of the multidisciplinary approach to cancer management, staging allows precision in documenting disease extent, thereby enhancing the quality of patient care. The recording of cancer stage at diagnosis is necessary to optimise patient care and provides a valuable means for recording patterns of disease presentation and monitoring advances in diagnosis and therapy. The objectives of staging described in the TNM staging system are as valid today as when implemented almost 50 years ago. PMID- 10470456 TI - The role of cancer staging in evidence-based medicine. AB - The classification of disease is a fundamental aspect of scientific medicine. Only when a specific condition has been recognized and characterized, can we start to acquire knowledge about it. Anatomic staging is a key element in the classification of malignant disease. The effectiveness of cancer treatment is almost always studied in groups of cases defined by stage. Clinical trials, therefore, create knowledge which is largely specific to particular stages of particular cancers. That knowledge can only be used to guide the practice of oncology if patients are staged. Staging the individual case permits the knowledge derived from previous experience to be used to guide the choice of treatment. The routine use of staging in a population also permits knowledge to be used to optimize the effectiveness of cancer control programs. PMID- 10470457 TI - Survey of Canadian doctors' attitudes to cancer staging. AB - OBJECTIVE: To collect opinions on the value of cancer staging, the impediments to realizing comprehensive staging and the support that may exist for providing stage information to cancer registries. DESIGN: The National Cancer Institute of Canada and the Canadian Committee on Cancer Staging have conducted an opinion survey. Opinions were sought about who should be responsible for allocating the stage, what criteria should be used to alter staging classification and whether population-based registries should contain comprehensive staging data. SETTING: The survey involved 1748 practising cancer specialists representing a cross section of Canadian physicians involved in the treatment and assessment of cancer patients. RESULTS: The overall response rate was 44.4%. Respondents gave broad support to the universal recording of cancer stage in Canada with 75.2% to 78.6% stating that staging is extremely useful for research, cancer statistics and caring for patients. Most respondents (98%) considered that all new cancer patients should have a specific stage assigned at the time of their first definitive treatment, and 78% believed that the clinician responsible for treating patients should determine the stage of cancer. A high proportion (73% to 81%) of respondents considered staging to be extremely important to define disease extent, make treatment decisions, estimate prognosis, facilitate consistent management and to compare results of treatment. The respondents consider lack of adequate information from pathology and imaging reports as barriers to staging. Only a small proportion of respondents were familiar with the organizations responsible for promoting and developing staging classifications. CONCLUSIONS: The Canadian survey showed strong support for universal staging and for the development of a central coordinated effort to promote cancer staging in Canada. PMID- 10470458 TI - The consultation to develop a national strategy for cancer staging in Canada. AB - The lack of uniform recording of stage is a continued barrier to the success of cancer control activities. To review the role of staging, in 1997 the Canadian Committee on Cancer Staging (CCCS) of the National Cancer Institute of Canada (NCIC) formalised a national Consultation initiative with the input of Canadian oncology leaders and international experts on staging. A document was created from prior background work by the Committee and pre-circulated for comment. The document described the importance of cancer staging to patient care, epidemiological and clinical research, and cancer control programs. The document was refined through an iterative process over much of a year which included a formal workshop of the participants and other meetings. The Consultation reaffirmed, with evidence, the central role of staging in all aspects of cancer management. The report of the Consultation, endorsed by the Board of Directors of NCIC, included the following recommendations: 1) the recording of TNM stage by the treating physician should be a standard of care for every cancer patient in Canada where TNM applies, 2) the Canadian Council on Health Services Accreditation should include the TNM stage for every cancer patient as a requirement for hospital accreditation. In conclusion, this process represents an effective method for raising awareness about important medical issues such as staging which are fundamental to the management and control of cancer. PMID- 10470459 TI - Canadian Council on Health Services Accreditation: client-centred cancer staging standards. AB - The Canadian Council on Health Services Accreditation has a mission to promote excellence in the provision of quality health care and the efficient use of resources in health organizations throughout Canada. The products and services of the CCHSA include national standards, onsite surveys and field education. The present standards of this voluntary, nonprofit national organization recognize the importance of cancer staging as an activity to evaluate the extent of disease in cancer patients and require cancer stage to be recorded. The CCHSA is implementing the next generation of accreditation, the Achieving Improved Measurement (AIM) project, which will lead to improved measurement of quality of health care. These quality-improvement efforts will emphasize the process of recording and reporting the TNM stage in all appropriate new cases of cancer. PMID- 10470460 TI - Issues in the implementation of cancer staging in Canada. AB - All sectors of the cancer control community in Canada agree that cancer staging is useful, and that stage should be assigned to every new case. At present, however, staging is not always recorded in the patient's records and treatment patterns and outcomes are rarely reported in terms of stage. This paper discusses what needs to be done to promote the use of staging in Canada. It is concluded that multifaceted programs of interventional continuing education (CE), tailored to meet the needs of the particular institution, offer the best prospect of success but the necessary organizational structure and information systems have to be put into place in advance. Implementation programs should be based on a thorough evaluation of the particular needs of the institution or community, and should be evaluated carefully in a few institutions before an attempt is made to disseminate them more widely. We recommend a phased approach to implementation which will first target institutions that already have the necessary infrastructure, i.e., provincial cancer centres. Demonstration of the feasibility and value of staging in that setting is seen as a means of promoting the adoption of staging in other institutions. PMID- 10470461 TI - Capturing tumour stage in a cancer information database. AB - OBJECTIVE: 1. To present the steps taken and lessons learned from one cancer centre's efforts to capture tumour stage information in a cancer database. 2. To determine the accuracy of the stage data through a chart audit. 3. To describe the potential uses of stage information in a cancer centre. DESIGN AND SETTING: This is a retrospective review of an initiative to capture tumour stage information at a regional cancer centre in Ontario. DATA SOURCES: The minutes of the centre's Health Records and Medical Advisory Committees related to staging were reviewed. Data on stage by tumour type was extracted from the centre's Oncology Patient Information System (OPIS). Three hundred and ninety charts were analysed to assess the accuracy of stage information and identify staging errors. Health Information Services workload statistics were reviewed to determine the types and frequency of projects undertaken using stage-related data. RESULTS: In January 1994, the Ottawa Regional Cancer Centre introduced policies and procedures to capture stage-related information. Standardized staging forms and a physician reminder system encouraged the centre's physicians to record tumour stage within 3 months of new patient registration. Of all qualifying cases in 1994, 92% were staged. A medical audit in 1998 of 390 charts from the 3 previous years of staging data revealed that 71.5% of the charts reviewed had been staged completely. Of the incompletely staged cases, 19% to 57% had TNM recorded, but the stage grouping was not recorded, or the "stage" was the extent of disease at the time of disease progression rather than at initial diagnosis (35% to 71%). Physician-related staging errors occurred in 2% to 5% of cases; data-entry errors occurred in 3% to 6% of cases. CONCLUSIONS: Stage information has enabled the centre to better describe its patient clientele for accreditation purposes and to assist researchers in estimating the number of patients potentially available for prospective and retrospective studies. It is being used to guide targeted educational initiatives to selected populations in the region's catchment area and assists administrators in estimating resource needs. Resistance to the capture of stage information can be overcome with persistence, the development of procedures that facilitate physician compliance, including a reminder system, the development of institutional policies and procedures and by feedback on the uses and availability of stage information. PMID- 10470462 TI - An international perspective on Canada's role in cancer staging. AB - Canadians have been involved in the issues of cancer staging since the inception of the international effort led by the UICC. The seminal roles of Sellers and Bush in furthering the development of the concept of staging and how it relates to a patient's prognosis have served as an example to others in Canada and elsewhere. Canadians have shown a pattern of multidisciplinary representation on staging bodies with epidemiologists, radiation oncologists and surgeons involved in cancer staging over the years. The scope of involvement has ranged from the NCIC Canadian Committee on Cancer Staging to the UICC, FIGO and the AJCC with its expert task forces. The recent consultation initiative to coordinate comprehensive nationwide staging by TNM is considered by the UICC as a potential model for other countries to follow. PMID- 10470463 TI - Management of ductal carcinoma in situ of the breast. Provincial Breast Cancer Disease Site Group. PMID- 10470464 TI - Clinical, angiographic, and procedural correlates of abrupt vascular closure during coronary intervention: a 10-year experience at Mayo Clinic. AB - A large matched-cohort study was carried out to determine correlates of in hospital abrupt vascular closure (AC). Univariate analysis identified current cigarette smoking (P = 0.021), myocardial infarction within 24 hr prior to procedure (P = 0.0035), emergency procedure (P = 0.02), lesion thrombus (P = 0.0001), and lesion angulation (P = 0.021) as significant clinical and angiographic variables. Relative to balloon angioplasty (PTCA), use of atherectomy (P = 0.015) and laser devices (P = 0.018) but not elective stent placement (P = 0.97) were associated with increased risk of AC. In the multivariate model, current cigarette smoking (P = 0.0474), lesion thrombus (P = 0.0001), lesion angulation (P = 0.0124), use of atherectomy devices (P = 0.001), and laser devices (P = 0.0037) remained as significant correlates of increased AC events. In conclusion, the risk of AC appears associated primarily with lesion characteristics and use of nonballoon devices other than stents. Elective stent placement did not appear to reduce AC risk over conventional PTCA; the small number of patients studied may have prevented any benefit from being observed. PMID- 10470465 TI - Abrupt postprocedural vascular closure: a new look at an old enemy. PMID- 10470466 TI - Novel distal occluder washout method for prevention of no-reflow during stenting of saphenous vein grafts. AB - This study assessed safety of the distal occlusion washout (DOW) method for prevention of no-reflow during stenting of degenerated saphenous vein grafts (SVGs). The DOW method involves protection of distal native coronary circulation with an occlusive balloon during the pretreatment and washout steps prior to stenting. Outcomes of stenting of 23 grafts in 21 patients after pretreatment with the DOW method were evaluated. The mean graft age was 7.4 +/- 4.3 years. The mean treated lesion length was 53 +/- 28 mm. Total occlusions were treated in 6 grafts and thrombotic lesions in 10 nontotally occluded grafts. One non-Q-wave MI and one acute stent thrombosis were observed. No deaths, Q-wave MIs, or subacute thromboses occurred. Follow-up in 18/21 (85.7%) patients at 28 +/- 8 weeks demonstrated target graft revascularization in 1 (5%) patient. The DOW method prevented clinically significant no-reflow in all 23 degenerated SVGs stented. PMID- 10470467 TI - Yes reflow. PMID- 10470468 TI - Predictors of diffuse-type in-stent restenosis after coronary stent implantation. AB - Diffuse-type in-stent restenosis (ISR) is associated with higher rate of restenosis after balloon angioplasty, requiring new therapeutic modalities; therefore, it is clinically important to identify the determinants of diffuse type ISR. We evaluate the clinical and angiographic variables to predict diffuse type ISR after coronary stent placement. Two hundred and ten ISR lesions in 196 patients (diffuse ISR, 114 lesions; focal ISR, 96 lesions) were reviewed in this study. Clinical, procedural and quantitative coronary angiographic parameters were analyzed. Diffuse-type ISR was defined as a > or = 50% lumen narrowing and > or = 10-mm length. Univariate analysis revealed that initial lesion length, smaller vessel size, diabetes, multivessel disease, multiple stents, and long stent were significantly associated with diffuse-type ISR. However, diabetes was the only independent predictor of diffuse-type ISR by stepwise multiple regression analysis (OR, 3.3; 95% CI, 1.4-7.4, P = 0.001). Diabetes was associated with diffuse-type ISR after coronary stent placement. It may reflect enhanced rate of neointimal hyperplasia within the stent in diabetic patients. PMID- 10470469 TI - Contribution of stenting to the results of rescue PTCA. AB - Failed thrombolysis in acute myocardial infarction (AMI) is associated with increased mortality. Controversial benefit of rescue percutaneous transluminal coronary angioplasty (PTCA) in these setting has been published. The feasibility, safety, and contribution of stenting to the outcome of AMI patients treated with this strategy is unknown. We studied the angiographic result and clinical outcome of 33 patients with failed thrombolysis referred for rescue angioplasty. Twenty three patients had stenting and 10 patients did not have stenting. Both groups had similar clinical and angiographic characteristics. Stent indications were nonoptimal result, 40%; bailout, 40%; elective, 20%. Angiographic success was 100% with stent vs. 91% with balloon alone (P < 0.8). Postprocedure residual stenosis was 1.5% (0%-10%) with stent vs. 18.05% (0%-30%) with balloon alone (P < 0.01). Thirty-day outcome with and without stent was mortality, 0% vs. 13% (P < 1.0); reinfarction, 10% vs. 0% (P < 0.30); target vessel revascularization, 0% vs. 21% (P < 0.21). The 6-month mortality was 0% with stent vs. 14% (P < 0.5). We conclude that stenting during rescue angioplasty is feasible, safe, and is associated with better immediate angiographic results. Although no obvious clinical benefit was found, a potential decrease in the revascularization rate was suggested. PMID- 10470470 TI - Predictors of stent thrombosis after primary stenting for acute myocardial infarction. AB - The aim of the study was to determine if a hypercoagulable state that may persist for several months after an acute myocardial infarction may contribute to an increased incidence of stent thrombosis. Primary stenting was performed in 104 consecutive patients with acute myocardial infarction using 147 coronary stents. Twenty-eight patients (27%) were diabetic and 55 patients (53%) were smokers. A single stent was placed in 63%, two stents in 33%, and more than two stents in 4% of the patients. Procedural success was obtained in 97% of the patients. All stents were deployed using high-pressure balloon inflation. The reference vessel diameter and minimal lumen diameter after stent deployment were 3.30 +/- 0.42 and 3.23 +/- 0.42 mm, respectively. Six patients (5.7%) developed stent thrombosis within 1 month after the procedure complicated by reinfarction in five of the six patients. At 1-month follow-up, all patients remained alive. On multivariate analysis, independent predictors of stent thrombosis were diabetes mellitus (relative risk [RR] 5.2; 95% confidence interval [CI] 1.8, 25.1), tobacco use (RR 4.5; 95% CI 1.3, 24.5), number of stents: 1 vs. > 1 (RR 3.7; 95% CI 1.1, 15.9), minimal lumen diameter poststent placement (RR 0.03; 95% CI 0.0002, 0.74), and duration of chest pain before intervention (RR 1.1; 95% CI 1.01, 1.25). Stent thrombosis had not been associated with diabetes mellitus and tobacco use previously but is in agreement with the enhanced platelet aggregability, coagulation factor abnormalities, and impaired fibrinolysis characteristic of these patients. PMID- 10470471 TI - Results of coronary stenting after delayed angioplasty of the culprit vessel in patients with recent myocardial infarction. AB - Little information is available concerning the effect of late coronary stenting in patients with recent myocardial infarction, especially long-term results. We retrospectively reviewed our results of 57 stent placements in 52 consecutive patients who received stents at an infarct-related lesion 24 hr to 30 days after an acute myocardial infarctions (median, 14 days). The average age was 67 years; 90% were male. Two patients who suffered from acute stent thrombosis received revascularization again and two early deaths were due to refractory cardiogenic shock before discharge. Mean patient clinical follow-up was 18.3 +/- 6.5 months. There were 1 subacute stent thrombosis, 1 cardiogenic death, and 10 patients (20.8%) in total suffering from angina class II to IV. Angiographic follow-up was performed in 36 patients (80%) at a mean of 7.5 +/- 3.1 months. Of these 36 patients, only 1 (3% of the total population undergoing follow-up angiography) had reocclusion at follow-up, but restenosis existed in 18 patients (50%). We conclude that there is still relatively high incidence of angiographic recurrence that is often silent in long-term follow-up, though the long-term result of late stenting in recent MI is low incidence of reocclusion. PMID- 10470472 TI - Short- and long-term outcomes after percutaneous transluminal coronary angioplasty in chronic hemodialysis patients. AB - The aim of this study was to obtain data on the outcomes of chronic hemodialysis patients who underwent percutaneous transluminal coronary angioplasty (PTCA). A retrospective chart analysis identified 31 such patients between August 1992 and October 1996. The mean follow-up period was 12.4 +/- 11.7 months. Angiographic success was achieved in 39 of 41 (95.1%) stenoses attempted. There were three in hospital deaths. Clinical success was achieved in 28 of 31 patients (90%). Two of the 28 survivors were lost to follow-up. Recurrent angina developed within 6 months in 14 of 26 patients (53.8%). Eleven and 17 of the 26 patients (42.3% and 65.4%) died within 6 and 14 months, respectively, after the PTCA procedure. Ten of the 17 deaths (58.8%) were due to cardiovascular events. Our study suggests that PTCA is technically feasible with high angiographic success rate in chronic hemodialysis patients. In-hospital mortality rate and rate of recurrent angina are high. Long-term prognosis is poor. PMID- 10470473 TI - Quantitative changes in reference segments during IVUS-guided stent implantation: impact on the criteria for optimal stent expansion. AB - Intravascular ultrasound is an established method to optimize stent implantation. Stent expansion is estimated from the relation between minimal in-stent cross sectional area and reference lumen area. We analyzed the periprocedural lumen increment in the reference segments and its impact on intravascular ultrasound (IVUS) criteria for optimized stenting. Seventy-five consecutive patients were studied with a 2.9 Fr, 30-MHz system and motorized pullback (0.5 mm/sec). Lumen area was measured by planimetry; absolute and relative differences in area (delta area) were calculated. Lumen area increment for reference segments proximal and distal to the stent was 6.4% +/- 10.3% and 6.1% +/- 10.8%; 49/75 patients fulfilled all IVUS criteria for optimal stent expansion at the final IVUS assessment, and 10/75 patients met all the IVUS criteria in relation to the first measurement of reference lumen area, but not in relation to the final measurement of reference lumen area. During high-pressure dilatation within the stent, reference lumen increment is visible. If reference lumen planimetry is not repeated after additional high-pressure balloon inflation, the final relative stent expansion may be overestimated. PMID- 10470474 TI - Emergency coronary artery bypass surgery following coronary angioplasty and stenting: results of a French multicenter registry. AB - This study investigates the influence of coronary stenting on the risk of emergency bypass surgery performed within 24 hr of percutaneous transluminal coronary angioplasty (PTCA) with particular concern for incidence and indication. Since 1995, coronary stenting has been increasingly performed in France during angioplasty procedures, altering significantly the role of emergency bypass surgery. The outcome of elective stenting and widespread use of coronary stenting and its influence on emergency surgery have not been evaluated so far. Through a retrospective (1995) and prospective (1996) registry, we analyzed the incidence, indication and results of emergency bypass surgery performed within 24 hr of PTCA in 68 and 57 centers, respectively, accounting for nearly half of all angioplasty procedures in France. Data were collected through questionnaires consisting of separate forms for every case report that were sent to every center. Over the two years, 26,885 and 27,497 procedures were investigated with a stenting rate of 46% and 64%, respectively. The observed need for emergency surgery was constantly low throughout this period (0.38% and 0.32%, respectively). Indications for surgery included complications directly due to stent in 37% of cases in the 2-year period. Outcome remained poor, with in-hospital mortality in 10% and 17% and myocardial infarction in 27% and 25% of cases, respectively. A comparison of the results in centers with and without surgical facilities showed no differences in outcome, despite a longer time to surgery (359 min +/- 406 min vs. 170 min +/- 205 min, P = 0.0001) and a lower incidence of emergency surgery (0.25% vs. 0.44%, P = 0.0001) in centers without on-site surgery backup. The French multicenter registry reveals an increase in the use of stents together with a dramatic decrease in the incidence of emergency bypass surgery (below 0.5%) following PTCA. There has been a significant evolution in the indication, and stent implantation now accounts for a third of the indications for emergency bypass surgery. PMID- 10470475 TI - Radiation exposure to children during coil occlusion of the patent ductus arteriosus. AB - The risks of excessive exposure to ionizing radiation are well described and measures are routinely taken to limit such exposure to both patient and personnel in the catheterization laboratory. Coll occlusion of the patent ductus arteriosus (PDA) as well as other more complex pediatric interventions has raised concern regarding radiation exposure, particularly as minimally invasive surgical techniques are being developed which lack such exposure risk. In eight consecutive patients, aged 0.7-7 years (median, 2.3 years), coil occlusion of a PDA was performed and surface entrance radiation dose determined by thermoluminescent dosimetry (TD). Total cumulative doses (PA + lateral dose) were also calculated for each patient. Entrance and cumulative dose was likewise measured in 12 patients undergoing standard diagnostic catheterization (DC) and in 5 consecutive patients undergoing pulmonary balloon valvuloplasty (PBV). The groups were comparable in age, weight, and body surface area (BSA). Total cumulative dose in the PDA patients was 97 +/- 25 mGy (mean +/- SE). There was no significant difference between the three groups in entrance dose absorbed at each location or in total cumulative dose. The mean total fluoroscopy time in the PDA occlusion group was significantly less than that of the PBV group (10.1 +/- 1.81 min vs. 19.3 +/- 2.29 min, P < 0.05) but was comparable to the DC group (13.2 +/- 1.5 min, P = NS). When the subjects were analyzed collectively, no correlation between fluoroscopy time and measured entrance dose was observed. The strongest correlates of total cumulative dose were patient weight (r = 0.67, P < 0.001) and BSA (r = 0.62, P = 0.001). Patients undergoing coil occlusion of a PDA are not exposed to increased radiation entrance dose compared to those undergoing standard DC and PBV. Furthermore, surface entrance radiation dose as determined by TD varies according to patient size for a given fluoroscopy time. PMID- 10470476 TI - Radiation risks to the patients and interventionalists: risk reduction. PMID- 10470477 TI - Long-term outcome of transcatheter coil closure of small to large patent ductus arteriosus. AB - Coil closure of patent ductus arteriosus (PDA) has become an accepted alternative to surgical closure in most pediatric cardiac centers. However, little is known about the mid-to long-term outcome of this procedure. Therefore, we evaluated the immediate, short-, and long-term outcome of transcatheter coil closure (TCC) of PDA using single or multiple Gianturco coils or the Gianturco-Grifka Vascular Occlusive Device (GGVOD). One hundred forty-nine patients underwent an attempt at TCC of their PDAs at a median age of 2.4 years (2 weeks to 55 years) and median weight of 13.5 kg (2.3-87 kg). There were 33 patients < 1 year of age. The median PDA minimal diameter was 2 mm (0.4-7 mm) with 26 patients whose PDA minimal diameter was > 4 mm. A 4 Fr catheter was used for coil deployment in 136 patients, a 3 Fr in 4, and an 8 Fr in 4 patients who received the GGVOD. A single coil was used in 77 patients and multiple coils (2-6) were used in 66 patients. One hundred forty-six patients had successful closure (142 had immediate complete closure and 4 had residual shunt), 3 patients failed the initial attempt (2 underwent surgical ligation and 1 had a successful second attempt a year later). Of the four patients with residual shunt, three underwent a second procedure with implantation of 1-3 coils resulting in complete closure in all and one patient had spontaneous resolution of the residual shunt. Complications were encountered in nine patients: six had coil migration with successful retrieval in four; two had left pulmonary artery stenosis (2.4 kg and 6.3 kg infants), and one patient had loss of femoral arterial pulse. The median fluoroscopy time was 16 min (2-152 min). One hundred forty-two patients had the procedure as an outpatient, five patients stayed greater than 24 hr, and two of these patients were in hospital for 1 month for noncardiac reasons. At a median follow-up interval of 3.0 years (1 month to 5.1 years), there were no episodes of delayed coil migration, delayed recanalization, thromboembolic episodes, or bacterial endocarditis. Lung perfusion scans performed at a median follow-up interval of 1.6 years in 31 patients who received multiple coils revealed 45% +/- 5% blood flow to the left lung. Long-term follow-up of coil closure of PDA indicates that the technique is safe and effective for most patients with PDA up to a diameter of 7 mm. PMID- 10470478 TI - Intraprocedural myocardial contrast echocardiography as a routine procedure in percutaneous transluminal septal myocardial ablation: detection of threatening myocardial necrosis distant from the septal target area. AB - Percutaneous transluminal septal myocardial ablation (PTSMA) has been introduced as an alternative to surgery for symptomatic hypertrophic obstructive cardiomyopathy (HOCM). Visualization of the ablation area prior to induction of the chemical necrosis is possible by intraprocedural myocardial contrast echocardiography (MCE). We report on two patients in whom MCE showed opacification of the medial papillary muscle or the left ventricular posterolateral free wall. In both patients the correct ablation area could be identified by MCE after a change of the target vessel, thus avoiding potentially fatal complications due to induction of a necrosis of myocardium distant from the septal target area. PMID- 10470479 TI - Upper posterior mediastinal tumor supplied by an atrial branch of the left circumflex artery. AB - We report a case of a 39-year-old woman with an upper posterior mediastinal tumor. The tumor was demonstrated by echocardiography and further defined by computerized tomography and magnetic resonance imaging. The tumor was fed by a large atrial branch of the left circumflex artery. Because of its location (adjacent to large vessels), it could not be resected by surgery. PMID- 10470480 TI - An overview of radiation safety regulatory recommendations and requirements. AB - This article reviews the formal basis of radiation protection programs. It also reviews the application of these programs in the cardiac catheterization laboratory. Radiation protection rules are based on epidemiology, radiobiology, and radiation physics. Applicable scientific results are periodically reviewed by committees reporting to the United States National Academy of Science and to the United Nations. National and international expert committees convert science to recommendations. The legislative process enacts enforceable regulations. These regulations need to be interpreted in relation to the actual risks experienced by the fluoroscopist. PMID- 10470481 TI - Ultrasound logic: the value of intracoronary imaging for the interventionist. PMID- 10470482 TI - Transluminal repair of an infrarenal aortoiliac aneurysm by a combination of bifurcated and branched stent grafts. AB - Transfemoral endovascular repair with a combination of bifurcated and branched stent grafts enables aortoiliac reconstruction with internal iliac perfusion preserved. We report a case of successful endovascular repair of aortoiliac aneurysm with use of a bifurcated and branched stent-graft. PMID- 10470483 TI - Of baling wire and bubble gum. PMID- 10470484 TI - Early clinical experience with the implantation of a novel synthetic coronary stent graft. AB - Coating stents with autologous venous grafts has been suggested to prevent problems associated with conventional stenting, but the need for surgical vessel harvest hampered broad application. A novel synthetic coronary stent graft (CSG) overcomes this limitation by a synthetic membrane, fixed between two thin metallic stents. We successfully implanted 21 CSGs in 18 patients for treatment of acute coronary rupture, thrombus-containing lesions, and lesions with plaque rupture or adjacent pseudoaneurysm. Substantial residual angiographic diameter stenoses were seen in seven CSGs (25% +/- 10% vs. 8% +/- 6%; P < 0.01), which were implanted with relatively small balloon catheters (balloon-to-artery ratio 1.00 +/- 0.09 vs. 1.24 +/- 0.18; P = 0.01) and required postdilatation. Overall, the largest balloon catheter applied measured 4.0 +/- 0.7 mm (balloon-to-artery ratio 1.21 +/- 0.20) and the inflation pressure was 16 +/- 3 atm. Final intravascular ultrasound imaging demonstrated adequate and symmetrical expansion of the CSG (> or = 85% +/- 15% of the reference lumen). Elective implantation was associated with two small non-Q-wave myocardial infarctions, resulting from unavoidable occlusions of side branches. Thus, implantation of CSG is feasible and safe. Adequate expansion can be achieved by the use of relatively large low compliant balloon catheters inflated with high pressure. PMID- 10470485 TI - Preliminary experience with the V-flex plus coronary stent: immediate and one month clinical outcome. AB - Procedural and 1-month outcome data following implantation of the V-Flex Plus stent in our first 54 consecutive patients (35 male; mean age, 62 years) are described. Sixty-four stents were implanted for 60 mainly complex lesions; 48% were left anterior descending; 20%, circumflex; 27%, right coronary artery; and 5%, saphenous vein graft. The Indication for stenting was elective in half of the patients and for a suboptimal result or as a bailout procedure in the other half. The stents were deployed at a mean of 12 atm (range, 6-18) and postdilatated to a mean of 15 atm (range, 8-20). Pre- and postdilatation balloon sizes were 2.96 +/- 0.57 mm and 3.16 +/- 0.34 mm, respectively. The procedural success rate was 98%. There were no deaths or Q-wave myocardial infarctions. One patient suffered a non Q-wave myocardial infarction and another developed a femoral false aneurysm. At 1 month follow-up, there were no additional events, in particular no revascularization procedures. Eighty-nine percent of patients were free of angina. Implantation of the V-Flex Plus stent is safe and effective with an excellent early success rate comparable to that of published randomized trials and registries of carefully selected patients. PMID- 10470486 TI - Safety and efficacy of AVE gfx stent implantation via 6 Fr guiding catheters. AB - This prospective study was designed to determine the feasibility of AVE gfx premounted stent systems in combination with 6 Fr guides. Between 1 April and 12 August 1997, 230 patients underwent AVE gfx coronary stent implantation via 6 Fr guides. The radial approach was used in 146 patients (63.5%). In 230 procedures (293 lesions), 237 guiding catheters were used. A total of 331 AVE gfx stents were implanted, 1.4 per patient. Backup, opacification, and friction were considered good in 85.8%, 96.4%, and 76.7%, respectively. Slight and severe friction was felt during combined use of long (> or = 18 mm), large-sized (3.5 mm) stents and small-sized guiding catheters (inner diameter, 0.061-0.062"). The presence of a second protecting guidewire impaired passage of the AVE gfx stent, also in large 6 Fr guides (ID 0.064"). Of 331, 320 (96.7%) stents were successfully deployed at the initial attempt. Ten stents (3%) had to be retrieved. Six of these were successfully placed at a second attempt and three slipped off the balloon, all successfully retrieved from the patient's circulation. At 1 month, 217 patients (94.4%) were free of events. The AVE gfx stent is compatible with 6 Fr guiding catheters. Use of new-generation, large bore 6 Fr gc (> or = 0.064") is recommended. PMID- 10470487 TI - Integrating bits and pieces: synapse structure and formation in Drosophila embryos. AB - During the development of the nervous system, numerous neurons connect to form complex networks. In order to build a functional network each neuron has to establish contacts with appropriate target cells, and at these contacts synapses of the right quality and strength have to be formed. Gaining insight into the mechanisms underlying this complex development is an important step towards a better understanding of how the nervous system is formed and behaviour generated. One model system in which synapse formation can be studied at the morphological, physiological and molecular level is that of the fruitfly Drosophila, and insights gained from Drosophila embryos are reviewed here. The first part of this review deals with the neuromuscular junction as the best-known synaptic contact in Drosophila. It describes: (1) its structure, (2) mechanisms underlying the formation of the neuromuscular cell junction and the arborisation of the presynaptic terminal, and (3) our present understanding of signal-dependent and independent processes during synapse formation at the neuromuscular junction. The last part of this review deals with the question of how particular neurons can adopt specific synaptic properties, stating as an example the development of the neural lineage of NB7-3, which gives rise to two serotonergic neurons. PMID- 10470488 TI - Development of auditory brainstem circuitry. Activity-dependent and activity independent processes. AB - Despite its complexity, the neural circuitry in the auditory brainstem of vertebrates displays a fascinating amount of order. How is this order established in such a precise manner during ontogeny? In this review, we will summarize evidence for both activity-independent and activity-dependent processes involved in the generation of the auditory brainstem circuitry of birds and mammals. An example of activity-independent processes is the emergence of crude topography, which, most probably, is determined by molecular markers whose expression is genetically controlled. On the other hand, neuronal activity supports cell survival, affects dendritic and axonal growth, and influences fine tuning of maps. It appears that various types of neuronal activity, namely spontaneous versus acoustically evoked, bilateral versus unilateral, uncoordinated versus patterned, play a role during different aspects of development and cooperate with the activity-independent processes to ensure the proper formation of neuronal circuitry. PMID- 10470489 TI - Neuropeptide precursor processing detected by triple immunolabeling. AB - Peptides that play critical physiological roles are often encoded in precursors that contain several gene products. Differential processing of a polypeptide precursor by cell-specific proteolytic enzymes can yield multiple messengers with diverse distributions and functions. We have isolated SDNFMRFamide, DPKQDFMRFamide, and TPAEDFMRFamide from Drosophila melanogaster. The peptides are encoded in the FMRFamide gene and have a common C-terminal FMRFamide but different N-terminal extensions. In order to investigate the regulation of expression of FMRFamide peptides, we generated antisera to distinguish between the structurally related neuropeptides. We established a triple-label immunofluorescence protocol using antisera raised in the same host species and mapped the neural distribution of SDNFMRFamide, DPKQDFMRFamide, and TPAEDFMRFamide. Each peptide has a unique, nonoverlapping cellular expression pattern, suggesting that the precursor is differentially processed. Thus, our data indicate that D. melanogaster contains cell-specific proteolytic enzymes to cleave a polypeptide protein precursor, resulting in unique expression patterns of neuropeptides. PMID- 10470490 TI - Immunocytochemical evidence of plasticity in the nervous structures of the rat lower lip. AB - In this immunocytochemical study we investigated the distribution of nervous structures in the lower lip of adult rats. The region is characterized by a rich cutaneous and mucosal sensory innervation originating from terminal branches of the trigeminal system. Lower lip innervation was investigated by detection of the general neuronal marker protein gene product 9.5 (PGP 9.5) and the growth associated protein 43 (GAP-43), a neurochemical marker of neuronal plasticity. The entire neural network of both cutaneous and mucosal aspects was stained by the antibody to PGP 9.5. In particular, nerve fibers were observed in the submucosal and the subepithelial plexuses. Thin immunoreactive fibers were observed within the epithelial layers ending as free fibers or as fibers associated with immunopositive Merkel cells. Well-identified anatomical structures receiving sensory or autonomic innervation were also surrounded by PGP 9.5-ir nerve fibers, in particular, hair follicles, vibrissae, glands, and blood vessels. GAP-43-immunostained nerve fibers were observed in all these structures; however, they were generally less numerous than the PGP 9.5-immunoreactive elements. An equal amount of PGP 9.5 and GAP-43 immunoreactivity occurred, in contrast, in the subepidermal and the submucosal plexuses, or in the epidermis and the mucosal epithelium. The present results show that GAP-43 is normally expressed in the mature trigeminal sensory system of the rat. Skin and oral mucosa are characterized by continuous remodeling that may also involve the sensory nervous apparatus. Continuous neural remodeling, regeneration and sprouting may be the reason for the observed expression of GAP-43. PMID- 10470491 TI - Immunocytochemical localization of substance P neurokinin-1 receptors in rat gingival tissue. AB - The distributions of substance P (SP) and the neurokinin-1 receptor (NK1-R), the receptor preferentially activated by SP, were examined in rat gingiva by immunocytochemical methods with light and electron microscopy. SP-immunoreactive nerve fibers were located preferentially in the junctional epithelium (JE) but few in the other oral and oral sulcular epithelia. NK1-R immunoreactivity was found in the endothelial cells (capillaries and postcapillary venules underlying the JE). NK1-R-labeled and -unlabeled unmyelinated nerve fibers were located close to the blood vessels and partially or completely covered by a Schwann cell sheath. In the JE, labeled naked axons without Schwann cell sheaths were observed. Neutrophils and macrophages in the connective tissue underlying the JE and in the JE were also labeled with NK1-R. Furthermore, NK1-R was found in the JE cells. Basically, immunoreaction products for NK1-R were found throughout various cells (endothelial cells, neutrophils, and JE cells) at invaginations of the plasma membrane and in vesicular and granular structures that are probably endosomes and are found close to both the plasma membrane and the nucleus. This is a first report, demonstrating the presence of NK1-R in the gingival tissue in the normal nonstimulated condition. Furthermore, it is thought that SP may modulate the permeability of blood vessels beneath the JE, the production of antimicrobial agents in neutrophils, and the proliferation and endocytotic ability of JE cells through NK1-R. PMID- 10470492 TI - Corticotropin-releasing hormone-immunopositive nerve elements in apposition to chicken gonadotropin-releasing hormone I-containing perikarya in Japanese quail (Coturnix coturnix japonica) brain. AB - The neuroanatomic basis of how stress inhibits reproduction in birds is not understood. To address this question we used double-label immunofluorescence histochemistry to determine whether corticotropin-releasing hormone (CRH) immunoreactive (ir) neuronal elements contact chicken gonadotropin-releasing hormone I (cGnRH I)-ir somata in brains of Japanese quail. The double-label system used a sheep anti-cGnRH I primary antibody with a secondary antibody conjugated to dichlorotriazinylaminofluorescein dihydrochloride for green fluorescence and a rabbit anti-CRH antibody with a secondary conjugated to Texas Red for red fluorescence. Immunohistochemical (IHC) distribution of both peptides resembled that in previous reports using single-label IHC. In four areas of the quail brain in which CRH nerve fibers and cGnRH I somata co-occurred (bed nucleus commissural pallii, nucleus preopticus medialis, nucleus septalis lateralis and nucleus accumbens), numerous instances were found of CRH-ir nerve fibers or terminals in apposition to cGnRH I cell bodies. These interactions provide a potential neuroanatomic route by which CRH may directly inhibit the activity of cGnRH-I-containing neurons, thereby inhibiting gonadotropin output and halting or slowing the progression of reproductive cycles. It remains to be demonstrated by electron microscopy whether these interactions, which appear abundant by IHC, represent instances of synaptic contact, as has been demonstrated to occur in analogous areas in mammalian species. PMID- 10470493 TI - c-fos expression in the forebrain and brainstem of White Leghorn hens following osmotic and cardiovascular challenges. AB - In chickens, hyperosmolality and hemorrhage increase hypothalamic vasotocin (AVT) gene expression and stimulate the secretion of AVT from the posterior pituitary gland. In this study, c-fos expression was used to identify areas in the forebrain and brainstem of the domestic chicken that are activated following acute osmotic stress and hemorrhage-induced hypotension. Conscious hens were osmotically stimulated by administering a single intraperitoneal injection of 3 M NaCl (5 ml/kg). Urethane-anesthetized hens were bled to a mean systemic arterial pressure of 80-90 mm Hg and maintained at this blood pressure for 1 h with additional bleedings as required. In both studies, the expression of c-fos was determined in control and experimental birds by using Northern blot analysis and in situ hybridization analysis. Osmotic stress and hemorrhage-induced hypotension increased c-fos expression in the same brain regions. Prominent structures in the forebrain that expressed c-fos mRNA following acute osmotic stress and hemorrhage induced hypotension included the supraoptic nucleus and paraventricular nucleus and nuclei within the hypothalamus that are anterior and ventral to the third ventricle. In the chicken, this region includes the organum subseptale, the o. vasculosum laminae terminalis, and the nucleus septalis medialis. In the brainstem, following either injection of 3 M NaCl or hemorrhage-induced hypotension, increased c-fos expression was observed in the nucleus of the solitary tract, parabrachial nucleus, area postrema, and locus ceruleus. Thus, the chicken central nervous system appears to use shared neuronal circuitry to stimulate hypothalamic AVT release in response to disturbances in body fluid composition and decreases in either systemic blood pressure or volume. PMID- 10470494 TI - Choline acetyltransferase immunoreactivity of putative intrinsic primary afferent neurons in the rat ileum. AB - The colocalisation of choline acetyltransferase (ChAT) with markers of putative intrinsic primary afferent neurons was determined in whole-mount preparations of the myenteric and submucosal plexuses of the rat ileum. In the myenteric plexus, prepared for the simultaneous localisation of ChAT and nitric oxide synthase (NOS), all nerve cells were immunoreactive (IR) for ChAT or NOS, but seldom for both; only 1.6 +/- 1.8% of ChAT-IR neurons displayed NOS-IR and, conversely, 2.8 +/- 3.3% of NOS-IR neurons were ChAT-IR. In preparations double labelled for NOS IR and the general nerve cell marker, neuron-specific enolase, 24% of all nerve cells were immunoreactive for NOS, indicating that about 75% of all nerve cells have ChAT-IR. All putative intrinsic primary afferent neurons in the myenteric plexus, identified by immunoreactivity for the neurokinin 1 (NK1) receptor and the neurokinin 3 (NK3) receptor, were ChAT-IR. Conversely, of the ChAT-IR nerve cells, about 45% were putative intrinsic primary afferent neurons (this represents 34% of all nerve cells). The cell bodies of putative intrinsic primary afferent neurons had Dogiel type II morphology and were also immunoreactive for calbindin. All, or nearly all, nerve cells in the submucosal plexus were immunoreactive for ChAT. About 46% of all submucosal nerve cells were immunoreactive for both neuropeptide Y (NPY) and calbindin; 91.8 +/- 10.5% of NPY/calbindin cells were also ChAT-IR and 99.1 +/- 0.7% were NK3 receptor-IR. Of the nerve cells with immunoreactivity for ChAT, 44.3 +/- 3.8% were NPY-IR, indicating that about 55% of submucosal nerve cells had ChAT but not NPY-IR. Only small proportions of the ChAT-IR, non-NPY, nerve cells had NK3 receptor or calbindin-IR. It is concluded that about 45% of submucosal nerve cells are ChAT/calbindin/NPY/VIP/NK3 receptor-IR and are likely to be secretomotor neurons. Most of the remaining submucosal nerve cells are immunoreactive for ChAT, but their functions were not deduced. They may include the cell bodies of intrinsic primary afferent neurons. PMID- 10470495 TI - Expression of mRNAs for type-I collagen, bone sialoprotein, osteocalcin, and osteopontin at different stages of osteoblastic differentiation and their regulation by 1,25 dihydroxyvitamin D3. AB - We have used in situ hybridization to evaluate the effects of 1,25 dihydroxyvitamin D3 (1,25 (OH)2 D3) on the expression of mRNA for bone-matrix proteins and to determine whether mature osteoblasts respond differently to 1,25 (OH)2 D3 than younger, newly differentiated osteoblasts. Rat calvaria cells were cultured for 7, 12, 15, and 19 days to obtain a range of nodules from very young to very mature. At each time point, some cultures were treated with 10 nM 1,25 (OH)2 D3 for 24 h prior to fixation. In control cultures, type-I collagen mRNA was detectable in osteoblastic cells in very young nodules and increased with increasing maturity of the nodules and the osteoblasts lining them. The bone sialoprotein mRNA signal was weak in young osteoblasts, increased in older osteoblasts, and decreased in mature osteoblasts. Weak osteocalcin and osteopontin signals were seen only in osteoblasts of intermediate and mature nodules. 1,25 (OH)2 D3 treatment markedly upregulated osteocalcin and osteopontin mRNAs and downregulated mRNA levels of bone sialoprotein and, to a lesser extent, type-I collagen in both young and mature osteoblasts. However, a marked diversity of signal levels for bone sialoprotein, osteocalcin, and osteopontin existed between neighboring mature osteoblasts, particularly after 1,25 (OH)2 D3 treatment, which may therefore selectively affect mature osteoblasts, depending on their differentiation status or functional stage of activity. PMID- 10470496 TI - Differential expression of CCAAT enhancer binding protein family in rat alveolar epithelial cell proliferation and in acute lung injury. AB - Although alveolar reorganization after acute lung injury depends on regeneration of alveolar epithelial cells, there is little knowledge of regulation of pulmonary healing process. Transcription factors may play key roles in this regulation. To investigate whether the CCAAT enhancer binding protein (C/EBP) family, alpha, beta, and delta, were involved in alveolar reorganization after injury, we examined expression of C/EBP proteins and mRNAs in lung injuries induced by lipopolysaccharide (LPS) or bleomycin (Bleo) and in cell proliferation by keratinocyte growth factor (KGF). By immunohistochemistry, we demonstrated that C/EBP alpha and C/EBP beta were expressed in alveolar type II cells and alveolar macrophages, but C/EBP delta was expressed restrictedly in some of alveolar type II cells in a spatial pattern in the control lungs. Further, these three C/EBP family members were differentially expressed in alveolar cell proliferation and in acute lung injury, in which, interestingly, C/EBP alpha and C/EBP delta were reciprocally expressed in alveolar type II cell proliferation and in pulmonary fibrosis. However, expressions of their mRNAs by in situ hybridization were dramatically increased in the affected lesions of the lungs by LPS and Bleo, and Northern blot analysis showed an increased abundance of the mRNA for C/EBP beta in LPS-treated lungs and for C/EBP delta in Bleo-treated lungs, compared with those in the control lungs. Thus, differential expression of the C/EBP family may be required to maintain and reorganize the basic integrity of alveolar structure during pathological states, which suggests an important role for the C/EBP family in maintaining normal alveolar architecture and function and in repairing the damaged epithelium after injury. PMID- 10470497 TI - Developmental regulation and ultrastructure of glycogen deposits during murine tooth morphogenesis. AB - The distribution and ultrastructure of glycogen deposits were investigated in the murine tooth germ by histochemical periodic acid-Schiff (PAS) staining and transmission electron microscopy. Lower and upper first molars were examined in mouse embryos at embryonic days 11.5-17 (E11.5-E17) and in 2-day-old postnatal (P2) mice. The oral and dental epithelia and the mesenchymal cells were generally PAS-positive during tooth morphogenesis. PAS-negative cells were present at E13 in the distal tip of the tooth bud epithelium and in the contacting mesenchyme, and this complete lack of PAS reactivity continued in the dental papilla mesenchyme and inner enamel epithelium during the cap and bell stages. The lack of glycogen deposits in the interacting epithelium and mesenchyme during early morphogenesis may be associated with their demonstrated high signaling activities. Mesenchymal cells in the dental follicle consistently possessed small clusters or large pools of glycogen, which disappeared by P2. Since an intense PAS reaction was seen in mesenchymal cells at future bone sites, the glycogen in the dental follicle cells may be associated with their development into hard tissue-forming cells. Ultrastructural observation of the enamel organ cells from the cap to early bell stages (E14-E15) revealed the occurrence of glycogen pools, which were associated with the Golgi apparatus and with vesicles having amorphous contents. Glycogen particles were also occasionally present inside vesicles or in the extracellular matrix. These may be associated with the exocytosis of glycosaminoglycan components into extracellular spaces and the formation of the stellate reticulum. PMID- 10470498 TI - Expression of axolotl RNA-binding protein during development of the Mexican axolotl. AB - Amphibians occupy a central position in phylogeny between aquatic and terrestrial vertebrates and are widely used as model systems for studying vertebrate development. We have undertaken a comprehensive molecular approach to understand the early events related to embryonic development in the Mexican axolotl, Ambystoma mexicanum, which is an exquisite animal model for such explorations. Axolotl RBP is a RNA-binding protein which was isolated from the embryonic Mexican axolotl by subtraction hybridization and was found to show highest similarity with human, mouse, and Xenopus cold-inducible RNA-binding protein (CIRP). The reverse transcriptase polymerase chain reaction (RT-PCR) analysis suggests that it is expressed in most of the axolotl tissues except liver; the expression level appears to be highest in adult brain. We have also determined the temporal and spatial pattern of its expression at various stages of development. RT-PCR and in situ hybridization analyses indicate that expression of the AxRBP gene starts at stage 10-12 (gastrula), reaches a maxima around stage 15-20 (early tailbud), and then gradually declines through stage 40 (hatching). In situ hybridization suggests that the expression is at a maximum in neural plate and neural fold at stage 15 (neurula) of embryonic development. PMID- 10470500 TI - Primary culture of antennal mechanoreceptor neurons of Manduca sexta. AB - We have developed a primary cell culture system of antennal mechanoreceptor neurons from early-stage pupal sphinx moth Manduca sexta. Dissociated neurons from the moth antennae differentiated, grew and survived for several weeks in a conditioned culture medium. Bipolar neurons with soma diameters of 10-25 microns from the basal portion of the antennae could be positively identified as mechanoreceptor neurons, presumably derived from Johnston's organ, using a monoclonal antibody that recognizes neurofilaments in these neurons. The immunoreactivity was clear and specific from the first day after dissociation and became stronger during several days in culture. These neurons appeared healthy and showed normal whole-cell properties only a few days after plating. We found numerous mechanosensitive ion channels responding to both negative and positive pressures on the somata and neurites of differentiated neurons. This new culture system provides access to mechanoreceptor neurons that has never been possible before, allowing the use of both mechanical and electrical stimuli on neurons that are free from the accessory structures surrounding them in intact preparations. PMID- 10470499 TI - Androgen-induced changes in Leydig cell ultrastructure and steroidogenesis in juvenile African catfish, Clarias gariepinus. AB - The present report focuses on the mechanism(s) involved in the steroid-induced decrease of androgen production in immature African catfish testes that was observed in previous studies. Juvenile animals were implanted with Silastic pellets containing different 11-oxygenated androgens (11-ketotestosterone, KT; 11 beta-hydroxyandrostenedione, OHA; 11-ketoandrostenedione, KA), testosterone (T) or estradiol-17 beta (E2). Control groups received steroid-free pellets. Two weeks later, testis tissue fragments were either incubated with increasing concentrations of catfish luteinizing hormone (LH), or incubated with [3H] pregnenolone ([3H]-P5) or [3H]-androstenedione ([3H]-A). Tissue fragments were also prepared for the quantitative assessment of Leydig cell morphology. Most of the parameters studied were not affected significantly by implantation of E2. Implantation of all androgens inhibited both the basal and the LH-stimulated androgen secretory capacity in vitro. This was associated with a reduced size of the Leydig cells and loss of half of their mitochondria. The studies on the metabolism of tritiated steroid hormones indicated that steroidogenic steps prior to 11 beta-hydroxylation, probably C17-20 lyase activity, were affected by all androgens. Although the effects of 11-oxygenated androgens and T on Leydig cells were mostly similar, previous work showed that only the 11-oxygenated androgens stimulated spermatogenesis, suggesting that distinct mechanisms of action are used by 11-oxygenated androgens and T. These mechanisms, however, seem to merge on the same target(s) to impair Leydig cell androgen production. Such a negative feedback mechanism may be of relevance in the context of the decline in androgen secretion per milligram testis tissue that accompanies the first wave of spermatogenesis in pubertal African catfish. PMID- 10470501 TI - Afferent projections of infrared-sensitive sensilla in the beetle Melanophila acuminata (Coleoptera: Buprestidae). AB - Beetles of the genus Melanophila are able to detect infrared radiation by using specialized sensilla in their metathoracic pit organs. We describe the afferent projections of the infrared-sensitive neurons in the central nervous system. The axons primarily terminate in the central neuropil of the fused second thoracic ganglia where they establish putative contacts with ascending interneurons. Only a few collaterals appear to be involved in local (uniganglionic) circuits. About half of the neurons send their axons further anterior to the prothoracic ganglion. A subset of these ascend to the subesophageal ganglion, and about 10% project to the brain. Anatomical similarities suggest that the infrared-sensitive neurons are derived from neurons supplying mechanosensory sensilla. The arborization pattern of the infrared afferents suggests that infrared information is processed and integrated upstream from the thoracic ganglia. PMID- 10470502 TI - An in vitro analysis of ecdysteroid-elicited cell death in the prothoracic gland of Manduca sexta. AB - The prothoracic glands of the tobacco hornworm, Manduca sexta, secrete the precursor of the insect molting hormone and normally undergo programmed cell death (PCD) during pupal-adult metamorphosis, between days 5 and 6 after pupation. This phenomenon can be elicited prematurely in vitro by the addition of 20-hydroxyecdysone (20E) to the gland cultures. To induce nuclear condensation in vitro in the glands from day-1 pupae, the effective dose range of 20E is 0.7-7 micrograms/ml and the minimum exposure period is 24 h. Prothoracic glands from different stages of pupal-adult development express different responsiveness to exogenous ecdysteroids. By utilizing terminal deoxynucleotidyl-transferase mediated dUTP nick-end-labeling (TUNEL) and the apoptotic DNA laddering method together with transmission electron microscopy, it has been demonstrated that the ecdysteroid-induced cell death of the prothoracic glands occurs via not only apoptosis but also autophagy, i.e., the induced dying cells show both severe nuclear fragmentation and autophagic vacuole formation, characteristics typical of apoptotic and autophagic cell death. The composite data indicate that ecdysteroids regulate directly both apoptotic and autophagic mechanisms of PCD of the prothoracic glands. PMID- 10470503 TI - Biosynthesis of major plasma proteins in the primary culture of fat body cells from the silkworm, Bombyx mori. AB - Plasma proteins termed "SP1" and "30K proteins" are synthesized by the fat body cells of the silkworm, Bombyx mori, in a sex- and stage-specific manner during larval development. We successfully established a primary culture of the fat body cells in order to investigate the regulatory mechanisms of plasma protein gene expression. The primary cultures of fat body cells contained at least two cell types: small oval cells, and large spherical cells. The cells adhered to and migrated on the cultured dish after plating. By the 7th day of cultivation, the cells clustered to form fat body-like structures, which were maintained for at least 3 months. Plasma proteins were actively synthesized in the primary cultures of the fat body cells isolated from the final instar larvae only when the cells tightly adhered to and clustered on the cultured dish. Immunocytochemical analysis revealed that only 10-15% of the clustered cells synthesized plasma proteins in our culture system, indicating that the primary culture comprises heterogeneous cells that are morphologically and functionally distinct. The patterns of SP1 syntheses in primary cultures faithfully reproduced their sex dependency in vivo. PMID- 10470504 TI - Anatomy, function, and pathophysiology of the posterior tibial tendon. AB - The posterior tibial tendon is vital for the structure and function of the foot and ankle. Dysfunction of the tendon can be debilitating and devastating. In recent years, much attention had been directed toward the diagnosis and treatment of PTTD. To properly diagnose and devise an appropriate treatment regimen, the anatomy, function, and pathophysiology associated with PTTD need to be thoroughly understood. PMID- 10470505 TI - Planar dominance. A major determinant in flatfoot stabilization. AB - For treatment of a symptomatic flat foot in a child or an acquired flatfoot from posterior tibial dysfunction in the adult, an understanding of the biomechanics and function of the foot and leg is essential to achieve an excellent outcome. Evaluation of supramalleolar deformity directly influences the procedure choice and eventual success of the procedure. An understanding of the level of deformity also indicates the procedure to be used. Subtalar and midtarsal joint function, although related, have their own primary source of deformity. Correction through the subtalar and midtarsal joints must be individualized. Augmentation of the primary repair must be performed when overall instability is present, such as an elongated or deformed posterior tibial tendon or medial ligamentous structure. Evaluation of the gross deforming force of the triceps suri also must be evaluated to provide and ensure predicted outcomes. PMID- 10470507 TI - Conservative management of posterior tibial tendon dysfunction, subtalar joint complex, and pes planus deformity. AB - Conservative management of PTTD can present from fairly simple to quite complex based on the wide range of clinical presentation that is inherent to this pathology. Treatment for PTTD ranges from the use of orthopedic footwear to the use of PTB AFOs, which certainly substantiates the prevalence of this disorder and the high rate of conservative management. With the increase in the population number and the fitness and health awareness that pervades our society, PTTD will more than likely continue to be a large part of the conservative footcare practitioner's practice in the future. The footcare team should include the pedorthist and orthotist to care for all the stages of PTTD. Perhaps with the increase in popularity of comfort footwear and increased use of orthoses (over the counter and custom-made), physicians can deter and alter the established pathway of the progression of this disorder. Preventive measures may, in some way, affect the foot health of the aging population. Conservative management of PTTD certainly has a place in today's healthcare climate. PMID- 10470508 TI - Treatment of posterior tibial tendon dysfunction with tendon procedures from the posterior muscle group. AB - The use of tendons from the posterior muscle group, specifically the FDL, as a means to repair PTTD is useful for the early stages of the deformity. Once the patient has reached the later stages and the foot becomes rigidly deformed with loss of the medial longitudinal arch, however, any attempt to reconstruct the area with tendon work alone fails. Tendon repair, tenodesis, and tendon transfer are attractive treatment options for PTTD, but care should be taken in choosing the correct patient for these procedures. Some authors note that side-to-side tenodesis does not address arch realignment. Other procedures combined with tendon work perhaps can help to reduce the shortcomings of isolated tendon procedures. Subtalar joint arthroeresis in combination with the tendon work seems to solve this problem. The authors have begun to explore this option and have performed this procedure on some patients. It is premature to address the effectiveness of this combined procedure. Similarly, tendon procedures augmented with other soft-tissue-type procedures also remains an option and is mostly ignored in the medical literature. Deland et al experimented with reconstruction of the spring ligament in a cadaver study, and believed that it should be considered in any reconstructive flatfoot surgery. Likewise, Myerson used some capsular reefing of the talonavicular joint in his tendon reconstruction to aid the correction of the forefoot-to-rearfoot relationship. The treatment of the patient with PTTD remains driven by the surgeon's preference, with little scientific research to guide him or her. There is much controversy regarding the efficacy of tendon procedures and the specific surgical technique of each procedure. Some variations may prove inconsequential, whereas others may prove revolutionary. The authors believe that the use of the tendon work as a means of treatment for PTTD is viable alone or in combination with other procedures. Much research is still needed to identify the best technique for each stage of the deformity. Wiekland has attempted to do this, but unfortunately has not offered any long-term follow-up to justify his treatment algorithms. Foot and ankle specialists should strive for clinical research, which allows better understanding of the appropriate treatment options for each progressive stage of PTTD. PMID- 10470509 TI - The Cobb procedure for stage II posterior tibial tendon dysfunction. AB - Flexor digitorum longus transfer or augmentation is currently the most popular adjunctive procedure for the repair of an attenuated or ruptured tibialis posterior tendon. Although the procedure is efficacious, an important functional muscle is sacrificed. Results show that similar results can be achieved with a tenodesis procedure by way of a split anterior tibial tendon repair. The authors have modified the Cobb procedure, and do not create a hole through the medial cuneiform or navicular. The thick, fibrous periosteal tissue at the medial aspect of the cuneiform is a sufficient tunnel for securing and positioning the tibialis anterior tendon. An additional site of healing and potential complications are avoided. The Cobb procedure is a useful and successful treatment option for PTTD, provides strong autograft augmentation to the posterior tibial tendon without sacrificing function of other tendons, and offers the surgeon and patient predictable outcomes with long-term satisfaction. PMID- 10470510 TI - Use of the Maxwell-Brancheau arthroereisis implant for the correction of posterior tibial tendon dysfunction. AB - Successful management of the flexible flatfoot deformity relies not only on correcting causal factors, but also on addressing the adaptive changes that have occurred and any coexisting deformities present. The three primary goals of any therapy--relief of pain, biomechanical control of excessive pronation, and prevention of the progression of the deformity--should be considered. With thorough assessment of causal factors and planal dominance of the deforming and compensative forces, one can expect to increase the predictability and success of the surgical management of PTTD in the flexible adult flatfoot deformity. PMID- 10470511 TI - Calcaneal osteotomy for the treatment of posterior tibial tendon dysfunction. AB - The surgical treatment of PTTD has been well-discussed in the literature. The calcaneal displacement osteotomy is a powerful, yet relatively easy, procedure that accomplishes a significant amount of frontal plane correction in the surgical treatment of PTTD. It can be performed as a straight medialization osteotomy for moderate hindfoot valgus, and must be performed with a resectional wedge in the treatment of large valgus angles. To achieve optimal correction, it must be coupled with procedures that address the triplanar nature of the deformity. When used in combination with other reconstructive procedures, it provides a valuable alternative to other more joint-destructive procedures. PMID- 10470512 TI - Talonavicular arthrodesis for correction of posterior tibial tendon dysfunction. AB - The awareness of PTTD has increased because of the efforts of McGlamry and Mueller. The treatment for PTTD depends on the patient's age and weight, systemic factors, length of time of the disease course, and the extent of foot collapse. The period of time from injury to diagnosis often is delayed because of the gradual progression of the condition. The patient that presents with an acute injury often responds well to a soft-tissue procedure. The delay in treatment usually necessitates the performance of an osseous procedure to correct the deformity and align the foot. The talonavicular arthrodesis is indicated in the flexible flatfoot deformity when degenerative changes of the subtalar joint are not present. The talonavicular arthrodesis is effective for correcting the flexible flatfoot deformity because it reduces the forefoot abduction, increases the height of the arch, stabilizes the medial column, and prevents excessive subtalar joint pronation. The primary complications associated with the talonavicular arthrodesis are nonunions and development of arthritis in adjacent joints. The incidence of nonunion can be directly attributed to poor surgical technique and early weight bearing during the postoperative period. The degenerative changes that occur in adjacent joints are often present preoperatively because of the long-standing valgus deformity. The procedure effectively maintains the correction of the flatfoot over a long period of time, and allows the patient to return to a pain-free lifestyle. The talonavicular arthrodesis is the procedure of choice in the flexible flatfoot deformity because the procedure corrects the malalignment of the subtalar and midtarsal joints and prevents excessive subtalar joint pronation. PMID- 10470513 TI - Triple arthrodesis and subtalar joint arthrodesis. For the treatment of end-stage posterior tibial tendon dysfunction. AB - The current literature clearly supports the use of subtalar and triple arthrodeses for the treatment of end-stage PTTD. There is debate, however, regarding whether or not an isolated fusion is preferable to the triple arthrodesis. Complete evaluation of the patient's deformity and symptoms is imperative before choosing to perform a rearfoot fusion. If the deformity can be isolated to the STJ, then perhaps a limited fusion is appropriate. With the close interrelationship of the subtalar and midtarsal joints, however, it is the authors' opinion that chronic dysfunction of the posterior tibial tendon infrequently causes isolated STJ pathology. Perhaps earlier intervention in the process of tendon degeneration, before multiple joint adaptations, would warrant an isolated fusion. We anticipate further research into the advantages of STJ and double arthrodeses over the triple arthrodesis. Clearer identification of the patients in whom these limited fusions are warranted is necessary, especially with respect to adult flatfoot secondary to PTTD. Currently, isolated and combined hindfoot fusions continue to be valuable salvage procedures in the treatment of end-stage arthritic deformities. PMID- 10470515 TI - Attention-deficit hyperactivity disorder on the frontlines: management in the primary care office. AB - Attention-deficit hyperactivity disorder (ADHD) is a common, disabling pediatric psychiatric disorder. Diagnosis of this disorder requires systematic data collection from multiple sources. The preferred therapy for ADHD is a combination of behavioral modification, educational assistance, and stimulant medications. PMID- 10470516 TI - Causes, diagnosis and treatment of pharyngitis. AB - Pharyngitis is the only respiratory tract infection in which microbiologic diagnosis has immediate influence on the treatment decision. Microbiologic diagnosis should always precede antibiotic treatment of pharyngitis to ensure optimal treatment. Penicillin administered for 10 days is still the drug of choice. PMID- 10470517 TI - The etiology and treatment of traumatic and septic shock. AB - Disseminated intravascular coagulation can cause multiple organ failure including adult respiratory distress syndrome by obstruction of visceral microcirculation by microclots. It was reasoned that if a clot causes vascular obstruction, lysing the clot by a plasminogen activator would be of value. PMID- 10470518 TI - Vulvovaginitis in healthy women. AB - Vulvovaginal symptoms are extremely common and result in millions of visits to practitioners' offices, STD clinics and emergency rooms. Vaginal infections or infectious vaginitis is responsible for only a minority of symptoms and is readily diagnosed. Epidemiology, diagnosis and therapy of vaginitis is reviewed. PMID- 10470519 TI - Recognition and treatment of arthritis in children. AB - Characteristics and differential diagnosis of juvenile rheumatoid arthritis (JRA) and diagnostic challenge of distinguishing it from juvenile spondyloarthropathy are reviewed. Although most children with limb pain do not have JRA, a reasonable approach is offered for those afflicted. PMID- 10470520 TI - The continuing challenge of exocrine pancreatic cancer. AB - Carcinoma of the exocrine pancreas continues to have one of the poorest outlooks of any cancer. Although worthwhile gains in survival may be achieved by early diagnosis and referral to a specialist surgeon, further improvements are unlikely without reliable screening techniques and improved adjuvant therapy. PMID- 10470521 TI - Risk factors for nephropathy in type 2 diabetes mellitus. AB - The main risk factors for diabetic nephropathy are hypertension, hyperlipidemia, and hyperglycemia. Nephropathy heralds the downhill course of arteriosclerosis. Early and intensive blood pressure and glucose control will attenuate the course of nephropathy and significantly reduce cardiovascular events and stroke. PMID- 10470523 TI - [Liver transplantation from a living donor]. PMID- 10470522 TI - Appendicitis in childhood. AB - Ultrasonography, CT scanning, and prolonged observation may improve diagnostic accuracy of appendicitis in children with atypical presentations. This article describes diagnostic pitfalls, including early presentation and abnormal appendiceal location. PMID- 10470524 TI - [The role of virologic tests in the treatment of chronic hepatitis C]. PMID- 10470525 TI - [Liver transplantation with a living related donor in the child]. AB - OBJECTIVES: Liver transplantation with living related donor has been recently developed to compensate for the insufficient number of liver grafts for children. The major problem is ethical because it implies voluntary mutilation of a healthy person. This paper report results in 37 living related donors. PATIENTS: Recipients were followed in Enfants-Malades Hospital. Investigations and donor surgery were performed at the Digestive Surgery Unit of Beaujon Hospital. RESULTS: One donor was re-operated for bleeding, and another one a biliary fistula treated with percutaneous drainage for one week. The post-operative course was uneventful in the other donors, with a follow-up of between 2 and 50 months. Thirty-three children are alive (90%), one of them underwent a second transplant for arterial thrombosis. Vascular and infectious complications, and the number of rejection episodes were the same as in transplantations with a deceased donor. Biliary complications were frequent (15 patients out of 37) and significantly increased morbidity. A teenage boy who received a small graft (0.9% of his weight) presented initially with hepatic insufficiency without encephalopathy. CONCLUSION: This technique has been shown to have a good balance between benefits and risks. Our experience confirms this, especially in very young children. Each case should be discussed individually and parental consent should be obtained without external pressure. Experience with this technique should be continued and at the same time the use of cadaveric grafts should be optimized. PMID- 10470526 TI - [Anatomic abnormalities in the papillary region and pancreaticobiliary pathology]. PMID- 10470527 TI - [International consensus conference on hepatitis C. Conclusions. The European Association for the Study of the Liver (EASL) Paris, February 26-28, 1999]. PMID- 10470528 TI - [Helicobacter pylori, gastritis and stomach tumors]. PMID- 10470529 TI - [Diagnostic value of high resolution sonography in Crohn's disease and ulcerative colitis]. AB - OBJECTIVE: To evaluate the diagnostic accuracy of high resolution sonography in patients with inflammatory bowel disease (MICI). PATIENTS AND METHODS: In patients with Crohn's disease (n = 48), ulcerative colitis (n = 23), indeterminate colitis (n = 3), inflammatory (n = 21) and non-inflammatory (n = 23) controls, high resolution sonography was performed and compared to colonoscopy (+/- retrograde ileoscopy) and/or baryum studies of the small bowel and the colon. RESULTS: Diagnosis of intestinal inflammation or not was correct in 69/74 MICI patients (sensitivity: 94.4%, specificity: 66.7%, global accuracy: 93.2%). Segment location was accurate in 58/74 (sensitivity: 80.3%, specificity: 66.7%, global accuracy: 79.7), more frequently in Crohn's disease, than in ulcerative colitis. Five out of six complications of Crohn's disease were diagnosed. In Crohn's disease, the method was more accurate in case of colonic or ileocolonic involvement. CONCLUSION: High resolution sonography is a reliable diagnostic tool for the detection of intestinal inflammation and related complications in MICI. In can be of value in the follow-up and seems particularly interesting in the case of temporary contraindication of invasive methods. PMID- 10470530 TI - Different gastritis features are linked to different gastric neoplasms. AB - OBJECTIVES: Helicobacter pylori infection induces gastritis, which may evolve to carcinoma or lymphoma. Whether duration of infection and inflammation pattern determine the outcome of the neoplastic process is not known. The aim of this study was to investigate the features of the gastritis associated with neoplasia. METHODS: Gastritis found in association with carcinoma (100 cases) and lymphoma (45 cases) were graded using the Sydney system. RESULTS: In particular in the antrum, gastric carcinomas, in particular of the intestinal type, were associated with a chronic (94%, n = 34/36) atrophic (92%, n = 33/36) gastritis and intestinal metaplasia (81%, n = 29/36). In diffuse type carcinomas inflammation was either absent or mild. An active (64%, n = 16/25), chronic gastritis (100%, n = 25/25) with lymphoid hyperplasia (72%, n = 18/25) was found in marginal zone cell lymphoma. CONCLUSIONS: Our study shows that the (pre)atrophic phases of inflammation are associated with gastric carcinomas. In contrast the active phase of inflammation, characterized by severe activity as well as severe chronicity, is found next to marginal zone cell lymphoma. PMID- 10470531 TI - Is there a link between the variation in gastric cancer mortality and differences in Helicobacter pylori prevalence in different regions of France? AB - An ecological study was performed to correlate the cumulative gastric cancer mortality rate to the prevalence of Helicobacter pylori infection in France. National data on mortality, standardized for age and gender, and the results of a nationwide prevalence study on Helicobacter pylori infection among 1,586 patients consulting for symptoms other than upper digestive tract symptoms, in seven defined French regions were used. The correlation was described by linear regression with the standardized data and then evaluated in a linear regression model including age and gender as co-variables. The Southwest region was the least affected by the infection (15.2%) while prevalence varied from 20.5 to 25.3% for the other regions. The cumulative gastric cancer mortality rate varied from 34.4 to 51.8/100,000. The prevalence of Helicobacter pylori infection in the model explained 5% of the variability in the gastric cancer mortality. A number of biases which were difficult to control could explain the lack of association between these variables. PMID- 10470532 TI - [Short chain fatty acids: effects on gastrointestinal function and therapeutic potential in gastroenterology]. PMID- 10470533 TI - [Hepatitis B virus reactivation after allogeneic bone marrow transplantation in a patient previously cured of hepatitis B]. AB - The presence of antibodies to HBs and HBc antigens indicates previous infection with hepatitis B virus but does not necessarily reflect viral clearance. Immunosuppression such as that observed in patients with bone marrow transplantation may be responsible for viral reactivation followed by acute exacerbation after withdrawal of immunosuppressive therapy. We report a case in a patient with natural immunity to hepatitis B who had undergone allogenic bone marrow transplantation with an identical sibling donor one year before for the chronic myelogenous leukemia in the first chronic phase. Ganciclovir treatment resulted in control of hepatitis virus B replication and in biochemical remission. We suggest that prevention relies on serological evaluation and therapy with active or passive immunisation or antiviral drugs in case of a rapid decline of anti-HBs Ab titers to undetectable levels. PMID- 10470534 TI - [Septic shock during the immediate post-partum period revealing a liver abscess in a woman with Crohn's disease]. AB - Liver abscess is a rare complication of Crohn's disease. We report one case in a 27 year-old woman, presenting as a sepsis after delivery. Crohn's disease was quiescent at presentation. Pregnancy was the only predisposing condition. An intestinal fistula was diagnosed by ultrasonography. The patient was treated by antibiotics, bowel resection and surgical drainage of the abscess. PMID- 10470535 TI - [Gastric granulocytic sarcoma revealed by a massive digestive hemorrhage]. AB - We report the case of a 37-year-old-man having a chronic myelogenous leukemia, who presented, one month after a splenic acutization, massive gastrointestinal bleeding from ulcerated nodules of the gastric fundus. The histologic examination of one of these nodules showed granulocytic sarcoma. In spite of an endoscopic treatment by sclerotherapy with adrenalined serum, the death occurred during a hemorrhagic recurrence. This observation, which is the third case reported of gastric granulocytic sarcoma during the acutization of a myelogenous chronic leukemia, and the first revealed by fatal gastrointestinal bleeding, shows the particular gravity of gastrointestinal bleeding complicating granulocytic sarcoma. PMID- 10470536 TI - [Primary chronic ulcerative jejunoileitis complicated by hemophagocytic syndrome]. AB - An observation of chronic ulcerative ileo-jejunitis associated with a hemophagocytic syndrome leading to death is described. It was not associated with coeliac disease. The hemophagocytic syndrome had no other etiology than chronic ulcerative ileo-jejunitis. The relations between these two disorders are discussed. PMID- 10470537 TI - [Growth of stellar angiomas during treatment with calcium inhibitors]. PMID- 10470538 TI - [Alithiasic cholecystitis and hemorrhagic dengue]. PMID- 10470539 TI - [Digestive system hemorrhage from duodenal invasion of a hepatocellular carcinoma: 2 cases]. PMID- 10470540 TI - [Esophageal cancer and Bazex' acrokeratosis: cutaneous development and parallel neoplastic development]. PMID- 10470541 TI - [Recurrent acute pancreatitis from the rupture of a solitary pancreatic hydatid cyst into Wirsung's canal]. PMID- 10470542 TI - [Gastrointestinal mucormycosis: a rare cause of digestive system hemorrhage]. PMID- 10470543 TI - [Microscopic colitis and Ticlid]. PMID- 10470544 TI - Preferences for equity in health behind a veil of ignorance. AB - Individual attitudes to distributions of life years between two groups in a society are explored by means of an experiment. Subjects are asked to place themselves behind a veil of ignorance which is specified in terms of risk (known probabilities) for some subjects and in terms of uncertainty (unknown probabilities) for some subjects. The latter is argued to be the appropriate interpretation of Rawls' notion. It is found that subjects exhibit convex preferences over life years for the two groups, and that preferences do not differ between the risk and the uncertainty specifications. PMID- 10470545 TI - Gender, heroin consumption and economic behaviour. AB - The study examines differences in consumption and economic behaviour among male and female heroin users. A sample of heroin injectors (n = 1834) was interviewed near the needle-exchange service in Oslo and information on consumption, prices, and income was recorded. The consumption pattern of the heroin addicts varies by gender, with females consuming relatively less alcohol and cannabis but significantly more heroin than their male counterparts. The finding of greater heroin consumption among women is surprising. By means of a switching regression model, price and income elasticities for heroin are estimated. Women appear to be more responsive than men to changes in prices. However, the elasticities also differ substantially by dealing status, and non-dealers are more price-responsive than dealers. Non-dealing males emerge with a higher income elasticity compared with their female counterparts, whereas female dealers seem to respond more to changes in income than do male dealers. PMID- 10470546 TI - A model for the Dutch pharmaceutical market. AB - This paper develops a general-equilibrium model for the Dutch pharmaceutical market. The model derives the behaviour of patients, pharmacists, drug producers and parallel-importers from microeconomic principles and recognizes the interactions between the various actors. The latter is important since the general-equilibrium effects are sometimes at odds with intuition from a partial equilibrium approach. A calibrated version of the model is used to evaluate the effects of various policy experiments on the bill for pharmaceutical care in the Netherlands. These experiments include an incentive policy to encourage cost conscious drug distribution, an increase of copayments for pharmaceutical care, a reduction of trade barriers for parallel-importers, and market entry by producers of generic substitutes. PMID- 10470547 TI - HMO selection and Medicare costs: Bayesian MCMC estimation of a robust panel data tobit model with survival. AB - The fraction of US Medicare recipients enrolled in health maintenance organizations (HMOs) has increased substantially over the past 10 years. However, the impact of HMOs on health care costs is still hotly debated. In particular, it is argued that HMOs achieve cost reduction through 'cream-skimming' and enrolling relatively healthy patients. This paper develops a Bayesian panel data tobit model of HMO selection and Medicare expenditures for recent US retirees that accounts for mortality over the course of the panel. The model is estimated using Markov Chain Monte Carlo (MCMC) simulation methods, and is novel in that a multivariate t-link is used in place of normality to allow for the heavy-tailed distributions often found in health care expenditure data. The findings indicate that HMOs select individuals who are less likely to have positive health care expenditures prior to enrollment. However, there is no evidence that HMOs disenrol high cost patients. The results also indicate the importance of accounting for survival over the panel, since high mortality probabilities are associated with higher health care expenditures in the last year of life. PMID- 10470548 TI - Women's participation in rural credit programmes in Bangladesh and their demand for formal health care: is there a positive impact? AB - Within the overall aim of poverty alleviation, development efforts have included credit and self-employment programmes. In Bangladesh, the major beneficiaries of such group-based credit programmes are rural women who use the loans to initiate small informal income-generating activities. This paper explores the benefits of women's participation in credit programmes on their own health seeking. Using data from a sample of 1798 households from rural Bangladesh, conducted in 1991 1992 through repeated random sampling of 87 districts covered by Grameen Bank, Bangladesh Rural Advancement Committee (BRAC) and Bangladesh Rural Development Board (BRDB), this paper addresses the question: does women's participation in credit programmes significantly affect their use of formal health care? A non unitary household preference model is suggested to test the hypothesis that women's empowerment through participation in these programmes results in greater control of resources for their own demand for formal health care. The analysis controls for endogeneity due to self-selection and other unobserved village level factors through the use of a weighted two stage instrumental variable approach with village level fixed effects. The findings indicate a positive impact of women's participation in credit programmes on their demand for formal health care. The policy simulations on the results of this study highlight the importance of credit programmes as a health intervention in addition to being a mechanism for women's economic empowerment. PMID- 10470549 TI - Non- and semi-parametric estimation of age and time heterogeneity in repeated cross-sections: an application to self-reported morbidity and general practitioner utilization. AB - Patterns of self-reported morbidity and general practitioner (GP) utilization exhibit complex age, sex and time heterogeneity. Underlying patterns are often obscured by data which are overly 'rough' because of noise associated with adjacent year fluctuations. In this paper we describe methods to obtain smoothed estimates of age, time and birth-cohort effects using data from the General Household Survey (GHS), covering the period 1984-1995/6 inclusive. The methods outlined offer powerful analytic tools to research complex profiles or trends, particularly over age or time. The relationships of the morbidity and GP utilization measures with age, sex and survey year characteristics are estimated non-parametrically using roughness penalized least squares (RPLS). A semi parametric extension of this model is used to estimate the effect of the morbidity variables on GP utilization. Tests are employed for various forms of age and time heterogeneity including birth-cohort effects. Linear age specifications are rejected for all variables and evidence is found of time heterogeneity in one of the morbidity measures--limiting long-standing illness (LS)--and GP utilization. The advantages of employing non- and semi-parametric estimations in the presence of complex relationships such as those observed for age and time profiles are discussed. Adoption of these techniques by applied econometricians working in health economics is encouraged. PMID- 10470550 TI - Health and mortality of the elderly: the grade of membership method, classification and determination. AB - With the aging of society, issues concerning the reform of the Dutch health care system are ranked high on the political agenda. Sensible reforms of the health care system for the elderly require a thorough understanding of the health status of the old and of its dynamics preceding death. The health status of the elderly is intrinsically a multidimensional and dynamic concept and a rich set of indicators is needed to capture this concept in its full extent. This feature of health requires techniques to reduce dimensionality as, in general, it is difficult to simultaneously handle all indicators in any economic analysis. In the first part of this paper we focus on methods that comprise these multidimensional measures into a limited number of indices. The Grade of Membership (GoM) approach introduced by Manton and Woodbury (Methods of Information in Medicine 1982; 21) is specifically designed to characterize the complex concept of health. The method simultaneously identifies all dimensions of the concept of interest and the degrees to which an individual belongs to each of these types (i.e. grades of membership). We apply the method to a set of 21 indicators from a rich database of the Longitudinal Aging Study Amsterdam (LASA). The individual degrees of involvement in the different health dimensions obtained from this method are used in subsequent analyses of health and mortality. PMID- 10470551 TI - Sensitivity of elasticity estimates for OECD health care spending: analysis of a dynamic heterogeneous data field. AB - Current econometric research on the determinants of health care spending is characterized by three serious shortcomings: a lack of attention to dynamics, a failure to deal with heterogeneity and a neglect of sensitivity testing. The latter shortcoming is a particular problem in an area where there is little theoretical guidance with which to judge the validity of estimates. This paper deals with these three problems by adopting recently developed techniques for analysing dynamic heterogeneous data fields containing non-stationary variables. This method allows for parameter heterogeneity across countries and is preferred to pooling with country specific effects, since the latter does not yield consistent estimates in this context, despite its common use in studies of health care spending. Long-run and short-run elasticity estimates are found to be sensitive to both model specification and sample composition. An attempt is made to compensate for the weak theoretical base by increasing the emphasis on statistical and econometric criteria for model evaluation, and by subjecting the results to rigorous sensitivity testing. PMID- 10470552 TI - Measuring the impact of health insurance with a correction for selection bias--a case study of Ecuador. AB - This article develops and uses methodologies to evaluate the impact of publicly financed health insurance programmes on the use of health care. Using univariate and bivariate probit estimation techniques, the study tests and corrects for endogeneity resulting from selection bias. Potential endogeneity arises from the choice to be insured, eligibility for insurance, and differences in individuals' health status. The setting for the study is the country of Ecuador. The General Health Insurance (GHI) programme, which primarily covers workers in the formal sector of the economy, is found to have a strong positive association with the use of curative health care after correcting for selection bias, but no significant effect on the use of preventive care. Individuals with severe illnesses who are eligible for GHI have a preference for private health care, and self-select out of the GHI programme. The Seguro Campesino Social (SSC) programme, directed at farming populations, has positive but insignificant associations with both curative and preventive care. PMID- 10470553 TI - Epidemiology of bacterial meningitis. AB - Bacterial meningitis caused by Haemophilus influenzae type B (Hib) has almost disappeared from the United States, Canada, and other countries that have implemented routine vaccination with Hib conjugate vaccines. The overall incidence of meningitis in these countries has declined by more than 50%, and the age distribution of susceptibility has shifted, so that the disease is now more common in adults than in children. Another new feature of the epidemiology of bacterial meningitis has been the occurrence of clusters of meningococcal disease. Such clusters have been school related, mainly in adolescents, and most clusters have been associated with a clone of group C, serotype 2a. The role of cigarette smoking as a risk factor for bacterial meningitis has been confirmed and adds urgency to the efforts to control smoking in adolescents and young adults. PMID- 10470554 TI - Pathogenesis of bacterial meningitis. AB - Bacterial meningitis is fatal in 5% to 40% of patients and causes neurologic sequelae in up to 30% of survivors. Much has been learned recently about the mechanisms that lead to brain injury during meningitis. Once bacteria have gained access to the central nervous system, their multiplication triggers a complex host response consisting of humoral and cellular immune mediators, reactive oxygen intermediates, matrix-metalloproteinases, and other host-derived factors. Alterations of the cerebral vasculature, with disruption of the blood brain barrier and global and focal ischemia, ultimately lead to functional and structural brain damage. This article reviews current concepts of the pathophysiology of bacterial meningitis and emphasizes possible therapeutic strategies to prevent its harmful consequences. PMID- 10470555 TI - Models of experimental bacterial meningitis. Role and limitations. AB - The seriousness of bacterial meningitis has encouraged the development of animal models that characterize complex pathogenetic and pathophysiologic mechanisms, provide evaluation of pharmacokinetic and antimicrobial effects of antibiotics (especially since the worldwide emergence of multiresistant bacteria), and establish new adjuvant treatment strategies (e.g., use of anti-inflammatory agents). The information obtained from an animal model depends on the site of inoculation. For example, using intranasal, intravenous, subcutaneous, or intraperitoneal inoculation, it is the bacterial and host factors that determine the development of bacteremia and the potential for a pathogen to invade the central nervous system that primarily are studied. In contrast, experimental models using direct inoculation into the cerebrospinal fluid can reliably produce lethal infections over a predictable time course. Furthermore, because adult animals will not reliably develop meningitis after intranasal or intraperitoneal challenge, infant animals are used. Because these models bypass the natural dissemination of bacteria from the intravascular compartment to the central nervous system, the pathogenesis is artificial. These models, however, are extremely useful for the study of pathogen and host factors leading to meningeal inflammation and resulting complications, and for evaluating potentially useful agents for treatment therapy. During the past decade, the design of clinical studies has been stimulated by findings obtained from these animal models. PMID- 10470556 TI - Clinical presentations, diagnosis, and prognostic factors of bacterial meningitis. AB - The clinical presentations of children and adults with bacterial meningitis have not changed over the past several decades, and a high index of suspicion remains critical for timely identification of infected patients. With the virtual disappearance of H. influenzae type B meningitis (Hib) in areas of the world where Hib conjugate vaccine is administered routinely, the utility of commercially available tests for rapid detection of bacterial polysaccharides has diminished. Detection of gene products of meningeal pathogens in cerebrospinal fluid or blood is still experimental. The prognostic findings of recent studies are not different from those previously described, despite advances in the supportive care of critically ill patients. PMID- 10470557 TI - Pharmacokinetics and pharmacodynamics of antibiotics in meningitis. AB - The penetration of antimicrobials into the CSF is dependent on lipid solubility, molecular size, capillary and choroid plexus efflux pumps, protein binding, and the degree of inflammation. Penicillins, certain cephalosporins, carbapenems, fluoroquinolones, vancomycin, and rifampin provide the highest ratios of CSF levels to the MBC for common infecting organisms. For beta-lactam antibiotics, it is the duration of time that CSF concentrations exceed the MBC that determines the rate of bactericidal activity. It appears that levels should exceed the MBC for more than 50% of the dosing interval. The peak/MBC and AUC/MBC ratios are important determinants of efficacy for aminoglycosides and fluoroquinolones. Once daily dosing of aminoglycosides is as effective as multiple-daily dosing regimens in experimental meningitis, probably because of drug-induced prolonged persistent effects. Fluoroquinolones do not produce as prolonged persistent effects and are slightly less effective when administered once daily. Although steroid use can reduce the penetration and decrease the bactericidal activity of some antimicrobials, such as vancomycin, in experimental meningitis, the clinical impact of steroid use in human meningitis is still unclear. PMID- 10470558 TI - Antimicrobial and anti-inflammatory treatment of bacterial meningitis. AB - Mortality and morbidity rates of bacterial meningitis are still unacceptably high, and thus, new, potent antimicrobial agents and adjuvant anti-inflammatory strategies are being evaluated to improve patient outcome. With the declining rates of Haemophilus influenzae type B infections, after the introduction of conjugated vaccines, research to find preventive measures for Streptococcus pneumoniae and Neisseria meningitidis infections is underway. In the meantime, scientific effort is being directed optimally to treat disease caused by multiresistant pneumococcal strains. PMID- 10470559 TI - Emergence of drug resistance. Impact on bacterial meningitis. AB - Antimicrobial resistance has emerged among the three major bacterial pathogens causing meningitis. Chloramphenicol resistance in the meningococcus recently has been described, and although intermediate penicillin resistance is common in some countries, the clinical importance of penicillin resistance in the meningococcus has yet to be established. Beta-lactamase-producing Haemophilus influenzae are relatively common, and chloramphenicol resistance is emerging. Third-generation cephalosporins are required to treat meningitis caused by these resistant strains. Pneumococcus resistance to penicillin and to chloramphenicol is widespread, and resistance to third-generation cephalosporins is found in many parts of the world. Correct management of these strains includes the addition of vancomycin or rifampin to therapy with third-generation cephalosporins. PMID- 10470560 TI - Supportive management in bacterial meningitis. AB - The central nervous system and systemic complications of bacterial meningitis cause significant morbidity and mortality. This article offers insight into the clinical features, pathogenesis, and management of these complications. In many instances, the improved outcome of intervention is based on clinical suspicion and early recognition. The management of complications is evolving and is presently based mainly on supportive care. PMID- 10470561 TI - Early management of meningococcal disease. AB - Meningococcal disease is increasing in incidence in many countries, and effective vaccines for serogroup B strains will not be available for at least 5 to 10 years. In the interim, it is attention to principles of good clinical practice, particularly in the early management of the disease, that have the potential to reduce by half the current case fatality rate of approximately 10%. As discussed in this article, those principles include increased awareness, understanding of the disease and its early symptoms by parents and healthcare professionals, and careful attention to the patient before admission and during the hospital stay. PMID- 10470562 TI - Prophylaxis of bacterial meningitis. AB - A comprehensive review of all major agents causing bacterial meningitis- meningococcus of the groups A, B, C, W135, and Y, pneumococcus, and Haemophilus influenzae type B (Hib)--is done in terms of preventing them by chemoprophylaxis or vaccination. Some evidence suggests that the group B meningococcal disease may also be very likely preventable by a vaccine that is already available. Excellent Hib conjugates use a technique that is expected to revolutionize immunoprophylaxis against most meningococcal and pneumococcal diseases in the near future. Unfortunately, the high cost of conjugate vaccines restricts their use in many poor countries. PMID- 10470563 TI - Bacterial meningitis and the newborn infant. AB - Bacterial meningitis in the neonate differs from meningitis in the older infant and child in a number of ways. Bacterial pathogens primarily are associated with the maternal genitourinary tract. Symptoms and physical findings may be nonspecific, and a high index of suspicion is needed. Management may vary depending on the maturity of the infant and the bacterial pathogen that is isolated. PMID- 10470564 TI - Nosocomial bacterial meningitis, including central nervous system shunt infections. AB - Nosocomial bacterial meningitis and CSF shunt infections result in considerable morbidity and mortality, necessitating an organized and thoughtful approach to prevention, diagnosis, and management. Prophylactic antibiotics appear to reduce the rate of postcraniotomy meningitis often caused by S. aureus. On the other hand, prophylactic antibiotics do not appear to reduce the risk of developing a CSF shunt infection. CSF shunt infections usually require shunt removal and antimicrobial chemotherapy to effect a successful outcome. PMID- 10470566 TI - Fever in the intensive care unit. PMID- 10470565 TI - NIPPV: patient-ventilator synchrony, the difference between success and failure? PMID- 10470567 TI - Molecular intensive care medicine. AB - The development of methods for the analysis of gene structure and function, referred to as recombinant DNA (deoxyribonucleic acid) technology, has created unprecedented opportunities for significantly improving the prevention or treatment of human diseases. Both practitioners working in this field and interested observers cannot fail to recognize that the remarkable progress in understanding disease pathogenesis has placed us on the threshold of a new, revolutionary era of clinical practice. In this context, molecular medicine--that is, the application of molecular biology to elucidating the causes and potential cures of disease, has become a major thrust of research at virtually all medical schools. Incorporating the techniques of molecular biology into the research arsenal of the physician should provide new opportunities to dissect out and define the reversible and irreversible intracellular processes giving rise to acute respiratory distress syndrome, sepsis, septic shock, or multiple system organ failure, the major causes of mortality in most intensive care units. PMID- 10470568 TI - Patient-ventilator asynchrony during noninvasive ventilation: the role of expiratory trigger. AB - OBJECTIVE: Air leaks around the mask are very likely to occur during noninvasive ventilation, in particular when prolonged ventilatory treatment is required. It has been suggested that leaks from the mask may impair the expiratory trigger cycling mechanism when inspiratory pressure support ventilation (PSV) is used. The aim of this study was to compare the short-term effect of two different expiratory cycling mechanisms (time-cycled vs flow-cycled) during noninvasive inspiratory pressure support ventilation (NIPSV) on patient-ventilator synchronisation in severe hypoxemic respiratory failure. STUDY POPULATION: Six patients with acute lung injury (ALI) due to acquired immunodeficiency syndrome (AIDS)-related opportunistic pneumonia were enrolled in the protocol. INTERVENTION: Each subject was first studied during spontaneous breathing with a Venturi oxygen mask (SB) and successively submitted to a randomly assigned 20' conventional flow-cycling (NIPSVfc) or time-cycling inspiratory pressure support ventilation (NIPSVtc). The pre-set parameters were: inspiratory pressure of 10 cm H2O, PEEP of 5 cm H2O for the same inspired oxygen fraction as during SB. A tight fit of the mask was avoided in order to facilitate air leaks around the mask. The esophageal pressure time product (PTPes) and tidal swings (delta Pes) were measured to evaluate the patient's respiratory effort. A subjective "comfort score" and the difference between patient and machine respiratory rate [delta RR(p-v)], calculated on esophageal and airway pressure curves, were used as indices of patient-machine interaction. RESULTS: Air leaks through the mask occurred in five out of six patients. The values of PEEPi (< 1.9 cm H2O) excluded significant expiratory muscle activity. NIPSVtc significantly reduced PTPes, delta Pes, and delta RR(p-v) when compared to NIPS-Vfc [230 +/- 41 (SE) vs 376 +/ 72 cm H2O.s.min-1; 8 +/- 2 vs 13 +/- 2 cm H2O; 1 +/- 1 vs 9 +/- 2 br.min-1; respectively] with a concomitant significant improvement of the "comfort score". CONCLUSIONS: In the presence of air leaks a time-cycled expiratory trigger provides a better patient-machine interaction than a flow-cycled expiratory trigger during NIPSV. PMID- 10470569 TI - A prospective study of fever in the intensive care unit. AB - OBJECTIVE: To determine the epidemiology of fever on the intensive care unit (ICU). DESIGN: Prospective, observational study. SETTING: Nine-bed general ICU in a 500-bed tertiary care inner city institution. PATIENTS: 100 consecutive admissions of 93 patients over a 4-month period between July and October 1996. INTERVENTIONS: All patients were seen and examined by one investigator within 24 h of ICU admission. Patients were followed up on a daily basis throughout their ICU stay, and all clinical and laboratory data were recorded during the admission. MEASUREMENTS AND RESULTS: Fever (core temperature > or = 38.4 degrees C) was present in 70% of admissions, and it was caused by infective and non infective processes in approximately equal number. Most fevers occurred early in the course of the admission, within the first 1-2 days, and most lasted less than 5 days. The median Acute Physiology and Chronic Health Evaluation (APACHE) II score was 15 (+/- 0.6). The 70 episodes associated with fever at any time were associated with a significantly higher APACHE II score on admission than the afebrile episodes (15.8 +/- 6.1 vs 12.1 +/- 6.7, p = 0.04). The most common cause of non-infective fever was in the group designated post-operative fever (n = 34). All the patients in the post-operative fever group were febrile on day 0 or day 1; their mean admission APACHE score was 12.4 (+/- 4.4) compared to 15.9 (+/- 7.1) for the remaining patients (p = 0.01). Fever alone was not associated with a higher mortality: 26/70 (37%) of febrile patients died, compared to 8/30 (27%) of afebrile patients, (chi 2 = 1.23, p = 0.38). Prolonged fever (> 5 days) occurred in 16 patients. In 13 cases, fever was due to infection, and in the remaining 3 both infective and non-infective processes occurred concurrently. The mortality in the group with prolonged fever was 62.5% (10/16) compared to 29.6% (16/54) in patients with fever of less than 5 days' duration, a highly significant difference (p < 0.0001). CONCLUSIONS: Fever is a common event on the intensive care unit. It usually occurs early in the course, is frequently non-infective and is often benign. Prolonged fever is associated with a poor outcome. Post operative fever is a well-recognised but poorly defined syndrome which requires further study. PMID- 10470570 TI - Aspiration of dead space allows normocapnic ventilation at low tidal volumes in man. AB - OBJECTIVE: Aspiration of dead space (ASPIDS) improves carbon dioxide (CO2) elimination by replacing dead space air rich in CO2 with fresh gas during expiration. The hypothesis was that ASPIDS allows normocapnia to be maintained at low tidal volumes (VT). DESIGN: Prospective study. SETTING: Adult intensive care unit in a university hospital. PATIENTS: Seven patients ventilated for neurological reasons were studied. All patients were clinically and haemodynamically stable and monitored according to clinical needs. INTERVENTIONS: ASPIDS implies that, during expiration, gas is aspirated through a catheter inserted in the tracheal tube. Simultaneously, a compensatory flow of fresh gas is injected into the inspiratory line. ASPIDS was achieved with a computer/ventilator system controlling two solenoid valves for aspiration and injection. RESULTS: At the basal respiratory rate of 12.6 breaths min-1, with ASPIDS VT decreased from 602 to 456 ml, as did the airway pressures to a corresponding degree. PaCO2 and PaO2 remained stable. At a frequency of 20 breaths min-1, with ASPIDS VT was further reduced to 305 ml with preserved normocapnia. ASPIDS did not interfere with the positive end-expiratory pressure (PEEP) level. No intrinsic PEEP developed. All patients remained stable. No haemodynamic or other side effects of ASPIDS were noticed. CONCLUSION: The results of this study suggest that ASPIDS may be a useful and safe modality of mechanical ventilation that limits alveolar pressure and minute ventilation requirements while keeping PaCO2 constant. PMID- 10470571 TI - The aetiology and pathogenesis of cardiopulmonary bypass-associated metabolic acidosis using polygeline pump prime. AB - OBJECTIVE: The pathogenesis of the metabolic acidosis of cardiopulmonary bypass (CPB) is not fully understood. New quantitative methods of acid-base balance now make it possible to describe it more clearly. Accordingly, we studied acid-base changes during CPB with polygeline pump prime and defined and quantified the factors which contribute to metabolic acidosis. DESIGN: Prospective cohort study. SETTING: Tertiary institution. PARTICIPANTS: 10 cardiac bypass graft surgery patients. INTERVENTIONS: Sampling of arterial blood at four time intervals: post induction, on CPB during cooling and rewarming, and at skin closure. Measurement of serum Na+, K+, Mg++, Ca++, Cl-, bicarbonate, and phosphate concentrations, arterial blood gases, and serum albumin, lactate, and pyruvate concentrations at each collection point. Analysis of findings according to quantitative physicochemical principles, including calculation of the strong ion difference apparent, the strong ion difference effective, and the strong ion gap (SIG). MEASUREMENTS AND MAIN RESULTS: All patients developed a mild metabolic acidosis. The median serum standard bicarbonate concentration decreased from 25.0 mEq/l post-induction to 22.3 mEq/l at cooling and 22.2 mEq/l at rewarming (p < 0.05). The standard base excess decreased from a median of 1.55 mEq/l prior to CPB, to 2.50 mEq/l at cooling, -1.65 mEq/l at rewarming and, -0.85 mEq/l at skin closure (p < 0.001). This mild metabolic acidosis occurred despite a decrease in the median serum lactate concentration from 3.20 mEq/l post-induction to 1.83, 1.80, and 1.58 mEq/l at the three other time points. The increase in the median serum chloride concentration from 104.9 mEq/l post induction to 111.0, 111.1, and 110.0 mEq/l at the subsequent time points (p < 0.0001) was the main cause of the acidosis. There was also a significant increase in the SIG of 3.8 mEq/l at cooling and rewarming (p < 0.0001), suggesting a role for other unmeasured anions (polygeline) in the genesis of this acidosis. CONCLUSIONS: Using quantitative biophysical methods, it can be demonstrated that, in patients receiving a pump prime rich in chloride and polygeline, the metabolic acidosis of CPB is mostly due to iatrogenic increases in serum chloride concentration and unmeasured strong anions (SIG). Its development is partially attenuated by iatrogenic hypoalbuminaemia. Changes in lactate concentrations did not play a role in the development of metabolic acidosis in our patients. PMID- 10470572 TI - The use of maximum SOFA score to quantify organ dysfunction/failure in intensive care. Results of a prospective, multicentre study. Working Group on Sepsis related Problems of the ESICM. AB - OBJECTIVE: To evaluate the performance of total maximum sequential organ failure assessment (SOFA) score and a derived measure, delta SOFA (total maximum SOFA score minus admission total SOFA) as a descriptor of multiple organ dysfunction/failure in intensive care. DESIGN: Prospective, multicentre and multinational study. SETTING: Forty intensive care units (ICUs) from Australia, Europe, North and South America. PATIENTS: Data on 1,449 patients, evaluated at admission and then consecutively every 24 h until ICU discharge (11,417 records) during May 1995. Excluded from data collection were all patients with a length of stay in the ICU less than 2 days following uncomplicated scheduled surgery. MAIN OUTCOME MEASURE: Survival status at ICU discharge. INTERVENTIONS: The collection of raw data necessary for the computation of a SOFA score on admission and then every 24 h, and basic demographic and clinical statistics. MEASUREMENTS AND MAIN RESULTS: Mean total maximum SOFA score presented a very good correlation to ICU outcome, with mortality rates ranging from 3.2% in patients without organ failure to 91.3% in patients with failure of all the six organs analysed. A maximum score was reached 1.1 +/- 0.2 days after admission for all the organ systems analysed. The total maximum SOFA score presented an area under the ROC curve of 0.847 (SE 0.012), which was significantly higher than any of its individual components. The cardiovascular score (odds ratio 1.68) was associated with the highest relative contribution to outcome. No independent contribution could be demonstrated for the hepatic score. No significant interactions were found. Principal components analysis demonstrated the existence of a two-factor structure that became clearer when analysis was limited to the presence or absence of organ failure (SOFA score > or = 3 points) during the ICU stay. The first factor comprises respiratory, cardiovascular and neurological systems and the second coagulation, hepatic and renal systems. Delta SOFA also presented a good correlation to outcome. The area under the receiver operating characteristic (ROC) curve was 0.742 (SE 0.017) for delta SOFA, lower than the total maximum SOFA score or admission total SOFA score. The impact of delta SOFA on prognosis remained significant after correction for admission total SOFA. CONCLUSIONS: The results show that total maximum SOFA score and delta SOFA can be used to quantify the degree of dysfunction/failure already present on ICU admission, the degree of dysfunction/failure that appears during the ICU stay and the cumulative insult suffered by the patient. These properties make it a good instrument to be used in the evaluation of organ dysfunction/failure. PMID- 10470573 TI - Sensitivity and specificity of a screening test to document traumatic experiences and to diagnose post-traumatic stress disorder in ARDS patients after intensive care treatment. AB - OBJECTIVE: Many survivors of critical illness and intensive care unit (ICU) treatment have traumatic memories such as nightmares, panic or pain which can be associated with the development of posttraumatic stress disorder (PTSD). In order to simplify the rapid and early detection of PTSD in such patients, we modified an existing questionnaire for diagnosis of PTSD and validated the instrument in a cohort of ARDS patients after long-term ICU therapy. DESIGN: Follow-up cohort study. SETTING: The 20-bed ICU of a university teaching hospital. PATIENTS: A cohort of 52 long-term survivors of the acute respiratory distress syndrome (ARDS). INTERVENTIONS AND MEASUREMENTS: The questionnaire was administered to the study cohort at two time points 2 years apart. At the second evaluation, the patients underwent a structured interview with two trained psychiatrists to diagnose PTSD according to Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) criteria. The reliability and validity of the questionnaire was then estimated and its specificity, sensitivity and optimal decision threshold determined using receiver operating characteristic (ROC) curve analyses. RESULTS: The questionnaire showed a high internal consistency (Crohnbach's alpha = 0.93) and a high test-retest reliability (intraclass correlation coefficient alpha = 0.89). There was evidence of construct validity by a linear relationship between scores and the number of traumatic memories from the ICU the patients described (Spearman's rho = 0.48, p < 0.01). Criterion validity was demonstrated by ROC curve analyses resulting in a sensitivity of 77.0% and a specificity of 97.5% for the diagnosis of PTSD. CONCLUSIONS: The questionnaire was found to be a responsive, valid and reliable instrument to screen survivors of intensive care for PTSD. PMID- 10470574 TI - Comparison of different methods for dead space measurements in ventilated newborns using CO2-volume plot. AB - OBJECTIVE: The aim of the study was to test the applicability of Ventrak 1550/Capnogard 1265 (V-C) for respiratory dead space (VD) measurement and to determine anatomic (VDana), physiologic (VDphys), and alveolar dead spaces (VDalv) in ventilated neonates. DESIGN: Prospective study. SETTING: Neonatal intensive care unit. PATIENTS: 33 investigations in 22 ventilated neonates; median gestational age 34.5 weeks (range 27-41), median birthweight 2658 g (range 790-3940). METHOD: The single-breath CO2 test (SBT-CO2) and transcutaneous partial pressure of carbon dioxide (PCO2) were recorded simultaneously and VD was determined (1) automatically (V-C software), (2) by interactive analysis of the PCO2 volume plot, and (3) manually by Bohr/Enghoff equations using data obtained by V-C. RESULTS: VD measurements were possible in all cases by method 3 but not possible by methods 1 and 2 in 22 of 33 investigations (67%), especially in preterm neonates, because of disturbed signals. V.Dana/kg (1.6 +/- 0.6 ml/kg, mean +/- SD), VDana/tidal volume (VT) (0.36 +/- 0.09) were lower compared to published data in spontaneously breathing infants, whereas VDphys/kg (2.3 +/- 0.9 ml/kg) and VDphys/VT (0.50 +/- 0.12) are comparable to data obtained from the literature. Five minutes after insertion of the sensor (dead space 2.6 ml) into the ventilatory circuit, the transcutaneous PCO2 rose above baseline for 3.2% (patients > 2500 g) and 5.7% (patients < 2500 g). The time necessary for one analysis was 50-60 min. CONCLUSION: In ventilated newborns, dead space measurements were possible only in one-third by SBT-CO2, but in all cases by Bohr/Enghoff equations. Improved software could further reduce the time needed for one analysis. PMID- 10470575 TI - The use of the laryngeal mask airway for inter-hospital transport of infants with type 3 laryngotracheo-oesophageal clefts. AB - Type 3 laryngotracheo-oesophageal clefts are rare congenital anomalies with a high mortality. In the past, transport of such infants to tertiary centres for surgical correction has proved extremely difficult, with the child's ventilatory status often deteriorating to such an extent that ultimate surgical intervention has not proved possible. We describe two cases of successful inter-hospital transfer of infants with type 3 laryngotracheo-oesophageal clefts using the laryngeal mask airway. PMID- 10470576 TI - Early postnatal dexamethasone for the prevention of chronic lung disease in high risk preterm infants. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of early administration of dexamethasone on the incidence of chronic lung disease (CLD) in high risk preterm infants and to evaluate the side effects of the early steroid administration. DESIGN: Randomised clinical trial. SETTING: Neonatal intensive care unit. PATIENTS: 50 infants at high risk of CLD were randomly assigned after 72 h of life to the dexamethasone group (n = 25) or to the control group (n = 25). The treated infants received dexamethasone intravenously from the 4th day of life for 7 days (0.5 mg/kg per day for the first 3 days, 0.25 mg/kg per day for the next 3 days and 0.125 mg/kg per day on the 7th day). The control group received no steroid treatment. RESULTS: The incidence of CLD at 28 days of life and at 36 weeks' postconceptional age was significantly lower in the dexamethasone group than in the control group (p < 0.001). Moreover, infants in the dexamethasone group remained intubated and required oxygen therapy for a shorter period than those in the control group (p < 0.001). Hyperglycaemia, hypertension, growth failure and mainly hypertrophy of the left ventricle were the transient side effects associated with early steroid administration. CONCLUSIONS: Early dexamethasone administration may be useful in preventing CLD, but its use should prudently be restricted to preterm infants at high risk of CLD. PMID- 10470577 TI - Severe brain injury in children: long-term outcome and its prediction using somatosensory evoked potentials (SEPs). AB - OBJECTIVE: To evaluate the outcome of children 1 and 5 years after severe brain injury (Glasgow Coma Score < 8) using a functional measure [Glasgow Outcome Scale (GOS)] and a health status measure (the Torrance Health State (HUI:1)) and to determine the ability of somatosensory evoked potentials (SEPs) to predict these long-term outcomes. DESIGN: Prospective study. SETTING: A 16-bed paediatric intensive care unit in a tertiary children's hospital. PATIENTS AND PARTICIPANTS: 105 children with severe brain injury. INTERVENTIONS: SEPs were recorded once in the first week after admission. Outcome was assessed 1 and 5 years after injury using the GOS and at 5 years after injury using HUI:1. MEASUREMENTS AND RESULTS: At 5 years, using the GOS, 46 (43.8%) children had a good outcome, 10 (9.5%) were moderately disabled, 2 (1.9%) severely disabled, 3 (2.9%) vegetative and 44 (41.9%) had died. At 5 years, 17 of 40 (42.5%) survivors from 1 year had changed outcomes: 12 had improved, 3 had worsened and 2 had died. For a normal SEP, positive predictive power was 85.4%, sensitivity 62.5%, specificity 87.8%, negative predictive power 67.2% and the positive likelihood ratio was 5.1. For bilaterally absent responses, positive predictive power was 90.9%, sensitivity 61.2%, specificity 94.6%; negative predictive power 73.6% and the positive likelihood ratio was 11.4. Outcomes using HUI:1 were: 30 (28.6%) had a good quality of life, 21 (20.0%) had a moderate quality of life, 7 (6.7%) a poor quality, 44 died (41.9%) and 3 (2.9%) survived in a state deemed worse than death. For a normal SEP, positive predictive power was 85.4%, sensitivity 68.6%, specificity 88.9%, negative predictive power 75.0% and the positive likelihood ratio was 6.2. For bilaterally absent responses, positive predictive power was 93.9%, sensitivity 57.4%, specificity 96.1%, negative predictive power 68.1% and the positive likelihood ratio was 14.6. CONCLUSION: The outcome for children with severe brain injury should be assessed 5 years after injury because important changes occur between 1 year and 5 years. Differences exist between outcomes assessed using the GOS and HUI:1 as they measure slightly different aspects of function. Consideration should therefore be given to using both measures. SEPs are excellent predictors of long-term outcome measured by either the GOS or the HUI:1. PMID- 10470578 TI - Bronchodilator responsiveness in a ventilator-dependent infant with severe tracheobronchomalacia. AB - A neonatal case of severe, ventilator-dependent tracheobronchomalacia (TBM) is described. The extent of the malacic segment was determined by endoscopy and tracheobronchography. Additionally, relevant and ever increasing reversible peripheral airway obstruction was documented by measuring the mechanical properties of the respiratory system before and after salbutamol. With the combination of endoscopically guided aortopexy and salbutamol infusion, the infant was eventually weaned from mechanical ventilation at the age of 86 days. We speculate that in ventilator-dependent infants with severe TBM the determination of bronchodilator responsiveness may have clinical consequences. PMID- 10470579 TI - Early but not delayed continuous arteriovenous hemofiltration improves cardiovascular function in sepsis in dogs. AB - OBJECTIVE: Continuous arteriovenous hemofiltration (CAVH) has been advocated as treatment to remove inflammatory mediators and thereby to improve hemodynamic parameters in sepsis. However, the results obtained with CAVH have been inconsistent. In a canine model of bacteremic Pseudomonas aeruginosa pneumonia, we tested the hypothesis that the time course of the institution of CAVH may be important in obtaining a beneficial treatment effect. METHODS: Two protocols were performed in phenobarbital-anesthetized dogs. In the early hemofiltration study (EHS), CAVH for 3 h was initiated 2 h post-pneumonia before mean arterial pressure (MAP) fell. In the late hemofiltration study (LHS), CAVH for 3 h was initiated at 5 h post-pneumonia when a decrease in MAP had already occurred. Hemodynamic measurements included cardiac output (CO), stroke volume (SV), and stroke work (SW). Myocardial depressant activity [filterable cardiodepressant substance (FCS)] found in plasma was assessed by bioassay at each measurement interval. RESULTS: In EHS, after 5 h of sepsis, SW, CO, and SV in the hemofiltered pneumonia group were higher as compared with the nonhemofiltered pneumonia group. In contrast, in LHS, no differences in hemodynamic parameters were found between the two pneumonia groups. In both EHS and LHS, plasma FCS activity was decreased to similar extents by CAVH. CONCLUSION: These results suggest the time course of institution of CAVH may be important in obtaining a beneficial treatment effect in sepsis. PMID- 10470580 TI - Management of severe and complicated malaria in the intensive care unit. AB - Malaria remains today one of the major health problems in the tropics with increased morbidity and mortality. The most serious complications are caused by Plasmodium falciparum, which, in contrast to the benign malarias, may progress to a life-threatening multi-system disease. Our case concerns a young woman in the 14th week of pregnancy, admitted to the ICU in a coma, with pulmonary oedema, haemolytic anaemia, renal failure and thrombocytopenia as complications of P. falciparum malaria. The case is discussed and possible explanations for the clinical picture and complications of P. falciparum malaria are given in the light of experiences from the literature. PMID- 10470581 TI - Pathophysiologic correlates of hypocholesterolemia in critically ill surgical patients. AB - OBJECTIVE: To assess correlates of hypocholesterolemia in moderate to critical surgical illness. DESIGN: Prospective analysis of laboratory and clinical data. SETTING: Department of surgery in a university hospital. PATIENTS: 135 patients undergoing uncomplicated abdominal surgery or with sepsis, liver failure, hemorrhage, severe cholestasis, or multiple organ dysfunction syndrome (MODS). INTERVENTIONS: Surgical and/or medical therapy according to clinical status. MEASUREMENTS AND MAIN RESULTS: Determinations of total cholesterol, additional variables, and clinical data. Cholesterol decreased after surgery, in sepsis, liver failure, acute hemorrhage, and MODS and increased in cholestasis. Hypocholesterolemia correlated with decreases in plasma proteins and indices of hepatic protein synthetic adequacy, with hemodilution from blood loss, and was moderated or prevented by cholestasis. CONCLUSIONS: These results help to explain the dynamics of the development, clinical relevance, and negative prognostic value of hypocholesterolemia in critical illness. PMID- 10470582 TI - Combination of inhaled nitric oxide and intravenous prostacyclin for successful treatment of severe pulmonary hypertension in a patient with acute respiratory distress syndrome. AB - OBJECTIVE: To investigate the combination of inhaled nitric oxide (iNO) and intravenously administered prostacyclin (i.v. PGI2) in a patient with severe pulmonary hypertension and acute respiratory distress syndrome (ARDS). DESIGN: Single case study. SETTING: Intensive care unit of a university hospital. METHODS: In an ARDS patient with severe pulmonary hypertension, gas exchange and hemodynamics were measured during combined treatment with iNO and i.v. PGI2. On two subsequent days, a protocol consisting of four 20-min periods was performed: baseline, 10 ppm iNO, 10 ppm iNO plus 4 ng kg-1 min-1, and 4 ng kg-1 min-1 PGI2 alone. At the end of each period hemodynamic and gas exchange data were obtained. RESULTS: The combination of iNO and i.v. PGI2 resulted in a marked decrease in pulmonary artery pressure and a concomitant increase in cardiac output which was more pronounced than the effect of either drug alone. During iNO, as well as during the combination of iNO and i.v. PGI2, oxygenation was improved, whereas during i.v. PGI2 alone oxygenation was worse than baseline. CONCLUSION: We conclude that the combination of iNO and i.v. PGI2 might be more useful than either drug alone when severe pulmonary hypertension leading to impaired right ventricular function is present in ARDS. A systematic study of this observation is warranted. PMID- 10470583 TI - ARDS after double extrinsic exposure hypersensitivity pneumonitis. AB - Hypersensitivity pneumonitis or extrinsic allergic alveolitis is a lung disease due to T cell and macrophage activation with IgA, IgG or IgE immunocomplex tissue lesions following extrinsic exposure to organic or inorganic agents. We report a case of hypersensitivity pneumonitis (pigeon protein sensitized) with a second nosocomial exposure to Aspergillus fumigatus proteins from a contaminated oxygen water humidifier: the second extrinsic exposure induced significant acute respiratory failure with ARDS. A pre-existing COPD syndrome requiring prolonged oxygen therapy (7 days) involved lung disease with delayed clinical diagnosis and therapy. Microbiological and mycological analysis of oxygen water humidifiers should be considered, especially for hypersensitivity pneumonitis patients, when a new inexplicable clinical impairment occurs. PMID- 10470585 TI - The Laryngeal Mask Airway: routine, risk, or rescue? PMID- 10470584 TI - A case of traumatic shock complicated by methamphetamine intoxication. AB - A case of a 38-year-old male with traumatic shock complicated by methamphetamine intoxication is presented. The patient was involved in an assault which resulted in cardiac tamponade and right ventricular outflow laceration. Pericardiocentesis was immediately performed. However, profound metabolic acidosis greatly in excess of that expected from the short duration of the shock was revealed by arterial blood gas analysis. Another cause of the metabolic acidosis was suspected. The patient subsequently admitted to intravenous use of methamphetamine. Following hemodynamic and metabolic stabilization by continuous pericardial drainage and intravenous administration of sodium bicarbonate, the patient underwent cardiac surgery. His postoperative course was uneventful. There is a substantial association between methamphetamine users and traumatic accidents. In such cases, early identification of drug use is important. Marked metabolic acidosis, which conflicts with the diagnosed cause of shock, may be a clinical clue to methamphetamine intoxication. PMID- 10470586 TI - Cranial nerve palsies and descending paralysis in a drug abuser resulting from wound botulism. PMID- 10470587 TI - Intracranial hypertension in head injury: management and results. PMID- 10470588 TI - Anaphylactoid reaction to pentastarch. PMID- 10470589 TI - Can dynamic PEEPi be greater than static PEEPi? PMID- 10470590 TI - Gastroenterology and internal medicine in Europe: united we stand. PMID- 10470591 TI - Early gastric cancer revisited. PMID- 10470592 TI - Correlation between endoscopic features of gastric antrum, histology and Helicobacter pylori infection in adults. AB - BACKGROUND: Helicobacter pylori induces histologic inflammation of mucosa variably correlated to different macroscopic features. Recent studies highlight that the presence of Helicobacter pylori could be assessed on the basis of the macroscopic pattern only, in particular nodularity. The present prospective study has correlated this and other endoscopic features, defined by Sydney classification of gastritis, both to Helicobacter pylori presence and histological patterns. RESULTS: Out of 532 patients, 364 were evaluable. The prevalence of Helicobacter pylori was 51.1% with a different distribution between the endoscopic features. Nodularity showed the highest positive predictive value in detecting the Helicobacter pylori presence (69.9%). The histological findings were: normal 26.9%, non atrophic gastritis 55.2%, atrophic gastritis 17.9%. There was a significant difference between abnormal endoscopic features in detecting the histologic gastritis, with endoscopic atrophy and nodularity showing the highest positive predictive value which reaches 96.7% and 91.8%, respectively. Helicobacter pylori infection and histologic gastritis were also present in 30.9% and 41.8%, respectively, of endoscopically normal subjects. Multivariate analysis has strictly correlated age with all abnormal endoscopic features, metaplasia with endoscopic atrophy, and chronic inflammation (gastritis) with nodularity. CONCLUSIONS: The single endoscopic features are poorly correlated with histologic changes and Helicobacter pylori status. Biopsies are mandatory in all cases. The causes of the different macroscopic findings are not yet fully understood. PMID- 10470593 TI - Quantitative antigliadin antibody measurement in clinical practice: an Italian multicentre study. SIGEP Working Group on Quantitative AGA Standardization. AB - BACKGROUND: The determination of class G and A serum antigliadin antibodies remains one of the most widely used screening tests for coeliac disease. The results from different laboratories are not always comparable, on account of changes in the technique and the different ways of expressing the results. AIMS: To: a) evaluate the physiological variation of serum antigliadin antibodies expressed in ng/ml, and b) establish the cut-off of quantitative antigliadin antibodies. PATIENTS: Patients were 127 individuals with active coeliac disease. Controls were 395 non-coeliac subjects (198 females and 197 males) aged 6 months to 45 years (median age: 4.9 years). METHODS: Antigliadin antibody enriched samples were obtained by affinity chromatography. The concentration of the eluted antigliadin antibodies was evaluated by nephelometry and enzyme-linked immunosorbent assay to establish a primary standard. An enzyme-linked immunosorbent assay for antigliadin antibody determination was run according to standard procedures. RESULTS: In controls, IgG-antigliadin antibody showed high variability in the 50th-90th centile range that peaked during the second year of life while IgA-antigliadin antibodies showed a lower variability and a less pronounced trend to decreasing values with age. A certain degree of overlapping between controls and coeliac patients was seen for both IgA- and IgG-antigliadin antibodies. The receiver operating characteristic analysis showed that the best discrimination was achieved by a cut-off of 8-10 ng/ml for IgA-antigliadin antibodies and 150-200 ng/ml for IgG-antigliadin antibodies. CONCLUSIONS: Antigliadin antibody concentration is not normally distributed and changes with age in non-coeliac subjects. The receiver operating characteristic analysis is a valuable tool for fixing the antigliadin antibody cut-offs between control and diseased individuals. The diagnosis of coeliac disease should always be confirmed by intestinal biopsy. PMID- 10470594 TI - Prevalence of gastro-oesophageal reflux in asthmatics: an Italian study. AB - BACKGROUND: Gastro-oesophageal reflux is regarded as an asthma trigger, its prevalence, in asthma, still being uncertain. AIM: Purpose of the study was to assess the prevalence of gastro-oesophageal reflux in adult asthma patients. PATIENTS: A series of 37 asthmatics were recruited consecutively over 6 months. Subjects were: 19 atopic (5 males; 38.0 years +/- 4.1 SE), and 18 non-atopic asthmatics (5 males; 53.2 years +/- 4.3 SE). They were well matched for sex and basal lung function, whilst mean age was appreciably higher (p < 0.007) in non atopic asthmatics. METHODS: Gastro-oesophageal reflux was detected by X-barium oesophagram, endoscopy, and 24 h oesophago-gastric pH-monitoring. RESULTS: Gastro oesophageal reflux was assessed in 78.9% of atopic, and in 83.3% of non-atopic asthmatics; the 24 h pH-monitoring was confirmed as the most sensitive method for gastro-oesophageal reflux detection. Hiatal hernia was found in 15.8% of atopic, and in 38.9% of non-atopic subjects. Respiratory symptoms always started long before gastro-oesophageal signs in atopic patients, while digestive symptoms came first (about 6 years) in 44.4% of non-atopic asthmatics. Oral theophylline was taken daily by 10.5% of atopic, and by 22.2% of non-atopic asthmatics. CONCLUSIONS: When non-atopic asthma starts at a later age without any family history of asthma, and/or after a long history of digestive symptoms, the role of gastro-oesophageal reflux can be strongly suspected. PMID- 10470595 TI - Asthma: reflux-induced or reflux-associated? PMID- 10470596 TI - Clinical and gastroscopic evaluation of amtolmetin guacyl versus diclofenac in patients with rheumatoid arthritis. AB - AIM: Amtolmetin guacyl (2-[2[1-methyl-5-(4-methylbenzoyl) pyrrol-2-yl] acetamido] acetic acid 2-methoxyphenyl ester) is a recently developed drug which, in preliminary studies, has shown effective anti-inflammatory properties with improved gastrointestinal safety. Our study was designed to investigate the efficacy and tolerability of amtolmetin guacyl 600 mg bid when compared to diclofenac 50 mg tid for 4 weeks. PATIENTS AND METHODS: A total of 64 patients aged 18-80 years, suffering from rheumatoid arthritis for more than 6 months and American Rheumatism Association functional class I, II or III were randomized in a double blind manner to amtolmetin guacyl or diclofenac for 4 weeks. Clinical and endoscopic evaluation were performed at baseline and at the end of the treatment. The mucosa was graded by means of a rating system emphasizing mucosal erosions. Only patients with endoscopy grade 0-1 entered the trial. RESULTS: The median post-treatment endoscopy injury scores were 0 (range 0-4) in the amtolmetin guacyl-treated patients and 2 (range 0-4) in the diclofenac-treated patients (p = 0.005). There were nine gastric ulcers: 1/32 (3%) in the amtolmetin guacyl group and 8/32 (25%) in the diclofenac group (p < 0.05; 95% confidence interval, -30-5%). 16/32 (50%) patients in amtolmetin guacyl group and 8/32 (25%) in diclofenac group had normal gastroduodenal findings (score = 0) (p < 0.05; 95% confidence interval, 5-50%). In patients with a history of peptic ulcer, a recurrence of gastric damage (score 3-4) was observed in 18% in the amtolmetin guacyl and in 53% in the diclofenac group (p < 0.05). The incidence of gastrointestinal symptoms did not differ in the two groups. Amtolmetin guacyl significantly reduced the number of swollen and painful joints, and the functional disability index; diclofenac significantly reduced the number of painful joints and the functional disability index score (p = ns). CONCLUSIONS: Amtolmetin guacyl effectively controlled the symptoms of rheumatoid arthritis, with very limited gastric toxicity. If these findings are confirmed on a wider scale, the drug might become a valid alternative to current therapies, especially for patients at risk, such as those with rheumatoid arthritis simultaneously requiring steroids and second-line drugs, or those with a history of peptic ulcer. PMID- 10470597 TI - A safe non-steroidal anti-inflammatory drug--the search continues. PMID- 10470598 TI - Multiple sclerosis and vaccination against hepatitis B: analysis of risk benefit profile. AB - BACKGROUND: Since 1994, the hypothesis of a potential causal relationship between vaccination against hepatitis B and multiple sclerosis (and other demyelinating diseases) was brought to the attention of the French health authority, and to public debate. In Italy, since 1991, vaccination against hepatitis B has been mandatory for newborns and 12-year-old children, and also recommended for high risk groups. AIM: To re-evaluate the risk/benefit profile of the Italian strategy of hepatitis B vaccination. SUBJECTS: The study population is a hypothetical cohort of 100,000 newborns. METHODS: We present a simulation of the hepatitis B cases that could be prevented with the vaccination and of the potential excess of multiple sclerosis cases which would occur, assuming different odds ratios of multiple sclerosis among vaccinees, and by effecting the vaccination at different ages. RESULTS: In the cohort, we would expect 1,099 hepatitis B cases, that would be prevented with vaccination. Assuming that the highest odds ratio of 1.7 reported is true, the excess of "life-time" multiple sclerosis incidence would be 0.3% for 12-year-old subjects, and 2.9% for adults. CONCLUSIONS: On the basis of these data, our opinion is that the hepatitis B vaccination strategy presently adopted in Italy for newborns, teen-agers and high risk groups should not be modified. PMID- 10470599 TI - The benefit-risk politics of vaccination against hepatitis B virus. PMID- 10470601 TI - The role of glutathione in the gastrointestinal tract: a review. AB - The primary role of glutathione is to protect cells from oxidative stress. It is abundantly distributed in the mucosal cells of gastrointestinal tract both in animals and man. The highest concentration is found in the duodenum. The amount of glutathione ingested with foods, age and drug or ethanol consumption affect glutathione concentration. The detoxifying capability of glutathione is directly related to its thiol group and to its function as a substrate for enzymatic activity; in fact, glutathione regulates the action of glutathione-peroxidases and glutathione-transferases. It has been documented that a direct relation between glutathione concentration and mucosal damage or between glutathione related enzymes and cancer occurrence is present in various pathological conditions of the gastrointestinal tract (from oesophagus to rectum). The present review underlines: a) the role of oxidative stress in numerous physiological and pathological conditions in experimental animals and man; b) the need to maintain a normal antioxidant potential in the mucosal cells of the gastrointestinal tract; and c) the possibility to evaluate, through clinical studies, how glutathione concentration, food intake, and gastrointestinal diseases are associated. PMID- 10470600 TI - Semiquantitative and qualitative assessment of hepatic iron in patients with chronic viral hepatitis: relation with grading, staging and haemochromatosis mutations. AB - BACKGROUND: Hepatic iron overload is a common but still poorly characterized finding in patients with chronic viral hepatitis. AIM: To evaluate lobular and cellular distribution of iron in patients with chronic viral hepatitis, the relation between hepatic iron distribution, grading and staging, and the frequency of haemochromatosis gene mutations. PATIENTS: Thirty-four patients with chronic viral hepatitis and iron overload; 34 matched chronic viral hepatitis controls without iron overload; 139 healthy controls. METHODS: Hepatic iron was assessed by hepatic iron concentration and Deugnier's score, histological grading and staging by Ishak's score, and frequency of haemochromatosis gene mutations by polymerase chain reaction-restriction assays. RESULTS AND CONCLUSIONS: Iron deposits were found in hepatocytes (94% of the patients), sinusoidal tracts (88%) and portal cells (59%). In 41%, iron deposits were homogeneously distributed in the hepatic specimen. Hepatocytic iron showed a decreasing gradient from Rappaport's zone 1 to 3. Heavy alcohol intake influenced the distribution rather than the amount of hepatic iron by increasing sinusoidal iron. Haemochromatosis gene mutations were more frequent in chronic viral hepatitis patients with iron overload than in those without iron overload and in healthy controls suggesting they contribute to pathogenesis of hepatic iron accumulation. The correlation between hepatic fibrosis and portal iron supports the fibrogenetic role of iron in chronic viral hepatitis. PMID- 10470602 TI - Review article: non-invasive methods for the diagnosis of Helicobacter pylori infection. AB - Non-invasive tests for the diagnosis of Helicobacter pylori infection are widely used in clinical practice and the management of patients with gastrointestinal disease. The serology and 13C-urea breath test are widely available, whereas salivary and faecal assay are still being developed. The non-invasive tests represent notable savings for the patient both in terms of money and discomfort. Whilst serology is the most widespread test its use is not advised in post treatment follow-up. The 13C-urea breath test is a simple, safe and highly accurate method ideal to evaluate, in the short-term follow-up, Helicobacter pylori eradication after treatment. PMID- 10470603 TI - Wilson's disease. AB - Wilson's disease is an autosomal recessive inherited disorder of copper metabolism resulting in pathological accumulation of copper in many organs and tissues. The Wilson disease gene is localized on human chromosome 13 and codes for a copper transporting P-type ATPase, -ATP7B. About one hundred mutations occurring throughout the whole gene have been documented so far. The most common is the His1069Gln point mutation. Wilson's disease may present under a variety of clinical conditions, the most common being liver disease (ranging from acute hepatitis, fulminant hepatic failure, chronic hepatitis, and cirrhosis), haemolytic anaemia, and neuropsychiatric disturbances. The diagnosis of Wilson's disease is usually made on the basis of clinical findings (Kayser-Fleischer rings, typical neurologic symptoms) and laboratory abnormalities (low serum caeruloplasmin, increased hepatic copper content). Molecular genetic testing is now the standard for testing asymptomatic siblings. Diagnosis in patients presenting with liver diseases is difficult and requires a combination of various laboratory parameters. Lifelong treatment with chelating agents (d-penicillamine, trientine) or with zinc is usually sufficient to stabilize the patient and to achieve clinical remission in most. Patients with advanced liver disease benefit from orthotopic liver transplantation. PMID- 10470604 TI - PAIR or PAI as percutaneous therapy of hepatic cystic hydatid disease: report of ten cases. Puncture, aspiration, injection, re-aspiration. PMID- 10470605 TI - Effect of tauroursodeoxycholic acid on ALT, GGT and viral load in patients with hepatitis C virus-related chronic hepatitis with reference to the genotype. PMID- 10470606 TI - An unusual cause of early TIPS dysfunction. PMID- 10470607 TI - Prevalence of duodenal bulb gastric metaplasia and Helicobacter pylori in patients with duodenal ulcer and normal duodenum. PMID- 10470608 TI - "Hansel and Gretel" technique for oesophageal intubation in patient with Zencker's diverticulum during endoscopic retrograde cholangio-pancreatography. PMID- 10470609 TI - [Head injuries in children: how to proceed without radiographs?]. PMID- 10470610 TI - [Evaluation, certification, accreditation: progress in radiology]. PMID- 10470611 TI - [A quality reference manual for radiology]. AB - The purpose of this article is to introduce a quality manual for radiology departments. A radiology department has implemented a quality improvement program since 1996. This manual was developed as a tool for quality improvement program. This manuscript, was based on foreign accreditation manuals as well as from French experiences and summarized. In addition, new criteria were added, especially in the field of interventional radiology. This reference book is first dedicated to the self-assessment of radiology departments. It can also be used for an external audit. PMID- 10470612 TI - [Quantitative angiography: update for peripheral vascular applications]. AB - Visual estimation of vessel diameter is unreliable and observer-dependent. With digital angiography, software for accurate quantification of arterial diameter are available. They allow: (1) selection of equipment of appropriate size (balloons, stents, etc) during revascularization procedures, (2) objective evaluation of the results of such procedures, (3) comparison between follow-up measurements and initial measurements and (4) provide reproducible data in the case of multicenter studies. The widespread availability of digital systems has been accompanied by a dramatic increase in the number of software packages for quantification. The present article reviews some of the currently available methods for quantification, presents the advantages and pitfalls of each system, and outlines their limits. PMID- 10470613 TI - [Cerebral angiography: a study of complications in 450 consecutive procedures]. AB - PURPOSE: To evaluate all types of complications, both minor and major, associated with modern cerebral angiography. MATERIALS AND METHODS: A prospective study of 450 consecutive cerebral angiographic procedures is reported. RESULTS: One patient (0.2%) died from a cholesterol embolus. In seven patients (1.6%), thromboembolic events occurred within 24 hours after the procedure, leading to transient ischemic symptoms in six and permanent hemiplegia in one. Two patients suffered from acute renal failure (0.4%). Transient cardiac arrhythmias were observed in three patients without consequence on the clinical outcome. Most complications of angiography occurred in patients referred from the neurology department for work-up of stroke syndrome. CONCLUSION: Our results show that morbidity and mortality rates related to the angiographic procedure did not decrease in spite of major improvement of angiographic materiel. Atherosclerosis is the main risk factor for complication. Most of the complications could be avoided by appropriate selection of indications and by using non-invasive techniques such as magnetic resonance angiography or helical CT angiography. PMID- 10470614 TI - [MRI symptomology in reflex sympathetic dystrophy of the foot]. AB - PURPOSE: To describe the MRI findings of reflex sympathetic dystrophy of the foot and ankle. METHODS: Retrospective study of 50 patients with reflex sympathetic dystrophy of the foot (5 with the cold form, and 45 with the warm form) diagnosed based on clinical and scintigraphic findings. All patients underwent MR imaging. The MRI findings were correlated with the clinical and scintigraphic findings. RESULTS: Patients with the cold form of reflex sympathetic dystrophy had no abnormality of signal at MR imaging. All patients with the warm from of reflex sympathetic dystrophy showed periarticular marrow edema at MR, typically involving more than one bone (mean of 4). Other findings were inconstant: soft tissue edema, joint effusion, and rarely, subchondral band of low T1W signal intensity of unclear etiology. CONCLUSION: MR imaging, including fat-suppressed T2W or STIR images and noncontrast T1W images, is helpful in patients with the warm or acute form of reflex sympathetic dystrophy of the foot. In patients with the cold form, MR imaging is helpful to exclude another underlying etiology for the symptoms and identify patients with the warm form of the process. PMID- 10470615 TI - [The exam request seen by the radiologist, the report seen by the clinician]. AB - PURPOSE: To analyze the impact of radiology requests and reports on the quality of the radiology consultation. METHOD: Over a 6 month period, the authors have collected radiology requests and reports deemed incorrect. These documents have been classified. A review of the literature has been conducted, then an attempt has been made to summarize the main elements to be included in radiology requests and reports to improve quality. RESULTS: Twelve categories of improper requests and 8 categories of improper reports have been identified. CONCLUSION: Requests and reports represent two essential steps of the consultation process in radiology. These two aspects are not sufficiently emphasized at medical school and during residency training in France. The efficiency of both steps mainly depends on the interaction between the referring physician and the radiology consultant. PMID- 10470616 TI - [Imaging of solitary maxillo-mandibular plasmacytoma]. AB - CT was performed in three patients with plasmacytoma of the maxilla and mandible. CT allows improved depiction of the tumor and associated local invasion and bony destruction but remains non-specific. Histology is mandatory for diagnosis and work-up necessary to confirm the solitary nature of the lesion. Intraosseous intramedullary lesions often evolve to multiple myeloma after a few years whereas extramedullary lesions, commonly involving sinonasal structures, typically remain solitary. PMID- 10470617 TI - [The persistence of a spheno-occipital synchondrosis in an adult]. AB - The postnatal development of the central skull base is a complex process: at least 25 separate ossification centers are assimilated in the maturing sphenoid and occipital bones. Some synchondroses may only be partially fused and persist during adulthood. We report the case of a 30-year-old man with a penetrating trauma of the central skull base. Skull computed tomography demonstrated a rare anatomic variant: incomplete fusion of the spheno-occipital synchondrosis. Knowledge of the normal skull base development and of its variants may prove helpful in differentiating a post-traumatic injury from a normal variant. PMID- 10470618 TI - [Massive benign pleural schwannoma]. AB - Schwannoma is a neoplasm of the peripheral nerve sheath that develops from schwann's cells. Pleural involvement is uncommon, and typically originates from an intercostal nerve of the posterior-superior mediastinum to then extend along the parietal and mediastinal pleura. We report a case of benign schwannoma of the right hemithorax, which we believe to be the largest ever reported at this time. The clinical presentation, acute respiratory failure, was unusual. The imaging findings of this tumor are described. PMID- 10470619 TI - [What is it? Acute small intestine occlusion due to primary malignant MALT-type lymphoma]. PMID- 10470620 TI - [Arterial lipiodol chemoembolization of a left liver metastasis through the right internal thoracic artery]. AB - The purpose of this article is to illustrate the efficacy of the chemoembolization in patients with hypervascular metastases and to describe the post-embolization change in vascularization pattern. Unusual collaterals may develop following embolization. A 59-year-old woman, followed for unresectable small bowel carcinoid tumor since 1991, underwent successful chemoembolization of several liver metastases. Only one liver lesion, located in segment IV, showed interval increase in size. This lesion was supplied by the right internal mammary artery. A branch of the right internal mammary artery was catheterized using a microcatheter and embolization was performed using doxorubicine-Lipiodol (Adriblastine, Lipiodol) and gelfoam (Spongel). No complications occurred after the procedure. The right internal mammary artery should be considered as a possible source of collateral arterial supply to the liver and should be evaluated in patients with local progression of disease. PMID- 10470621 TI - [The Bone Club. Meeting of 5 May 1999--Cochin Hospital, Paris]. PMID- 10470623 TI - Soluble interleukin-2R alpha test kits measure different epitopes: sIL-2R alpha test kits comparison with a newly developed inhouse sIL-2R alpha sandwich ELISA. AB - Cytokines are a group of low molecular weight glycoproteins important in cellular signaling for various responses such as activation and proliferation. They separate the cellular responses into Th1 and Th2 pathways, where each pathway releases its own cytokine group. Interleukins are an important section of the cytokine group, and consist of a number of members with their respective receptors such as interleukin-2 and interleukin-2 receptor (IL-2 and IL-2R, respectively). In this study, a soluble interleukin-2 receptor (sIL-2R alpha) sandwich ELISA was developed and tested with different samples. This method was compared with commercial test kits that measure sIL-2R alpha molecule. The results showed that different kits measure different epitopes on this molecule. PMID- 10470622 TI - Crossroads of the effects of cyclophosphamide pulse therapy for lupus nephritis- experience of 11 cases. AB - In this study time for initial assessment of monthly intravenous cyclophosphamide (CP) pulse therapy is discussed for a better outcome with less complications. Eleven patients with lupus nephritis (LN) resistant to conventional therapy (serum creatinine level < or = 2.7 mg/dl) were given 500 mg/m2 of CP 7-9 times with an interval of one month. Urinary protein (Up) decreased in all patients after 3 courses of CP pulse therapy and kept similar levels thereafter. In one group of patients (n = 7), Up decreased to < 2 g/day after 3 courses, while in the other group (n = 4), it did not decrease to < 4 g/day. Creatinine clearance increased by 0-100% in the former group, while it decreased by 5-20% in the latter group after 6-9 courses. Renal function of the patients with insufficient response after 3 courses tended to show no further improvement or worsened thereafter, although Up decreased during CP pulse therapy. A relatively small dose of CP (500 mg/m2) pulse therapy was useful in most LN patients regardless of the renal histology and it was thought important to assess its effects after 3 courses for a prediction of the clinical course. Modification of the protocol at that time might be necessary in regard to dose or interval of CP administration especially for patients with insufficient outcome. PMID- 10470624 TI - Comparison of lymphocyte subpopulations in the peripheral blood of patients with infectious mononucleosis and human immunodeficiency virus infection: a preliminary report. AB - The aim of this study was to analyse the lymphocyte subpopulations in the peripheral blood of HIV-1-infected adults to compare them with four patients with acute EBV infection. Lymphocyte subsets in 15 healthy controls, 40 HIV-1-infected adults and 4 EBV-infected patients with infectious mononucleosis were analysed by flow cytometry. The immunophenotyping of HIV-1-infected patients in different stages of disease showed a significant reduction in the percentage and absolute count of CD4+ T-lymphocytes, significantly increased percentage of CD8+ T lymphocytes, inverted CD4/CD8 ratio and an increase in the expression of activation marker HLA-DR compared to controls. The immuno-phenotyping profiles of HIV and EBV infection share some similarities as they both result in the decreased percentage of CD4+ T-lymphocytes, increased CD8+ T-lymphocytes and an inverted CD4/CD8 ratio. Patients with HIV infection could be distinguished from patients with EBV infection by the absolute lymphocytosis and increased expression of HLA-DR seen in the patients with infectious mononucleosis. In conclusion, both HIV-1 and EBV profoundly change the distribution of lymphocyte subpopulations in the peripheral blood. It is our opinion that flow cytometry could be an aid in the rapid distinguishing of patients with suspected primary HIV-1 infection from those with infectious mononucleosis (before serology data are available). PMID- 10470625 TI - Medical malpractice reform--the long view. PMID- 10470626 TI - Clinical assessment of malpractice case scenarios in an anesthesiology department. AB - STUDY OBJECTIVE: To determine whether anesthesiologists agree with each other when assessing actual malpractice clinical scenarios, whether their assessments comport with the actual malpractice verdicts, and whether they can accurately guess jury verdicts. DESIGN: Survey study requesting anesthesiologists to review 12 malpractice case scenarios, administered twice. SETTING: Academic medical center department of anesthesiology. MEASUREMENTS AND MAIN RESULTS: Mean interphysician agreement, mean Likert values assessing relative negligence, mean agreement with jury verdicts, mean success at predicting actual jury verdicts for 12 actual jury verdict case scenarios. Respondent anesthesiologists appeared homogeneous by training and years of experience. They showed high (> 80%) agreement among themselves in their assessments of the malpractice case scenarios over the two administrations (p = 0.13). In addition, mean Likert values as to relative negligence assessments by respondent anesthesiologists were not significantly different between administrations by case (p = 0.09 to 1.00). However, of the eight cases with complete or virtually complete agreement between respondent anesthesiologists, three (37.5%) disagreed with the verdict rendered by the actual juries. In addition, anesthesiologists showed significant disagreement (> 30%) among themselves in four of the case scenarios, indicating there may not be agreement regarding the standard of care in these clinical circumstances. Finally, anesthesiologists predicted jury verdicts poorly, with success rates of 50% or less in seven of the 12 case scenarios. CONCLUSIONS: For this sample of homogeneous anesthesiologists who demonstrated high clinical agreement, it appears that the malpractice system may not be able to function on its own terms in adjudicating malpractice claims. Although there was agreement among respondent anesthesiologists, these assessments were in direct opposition to actual verdicts, a significant percentage of cases resulted in disagreements as to the appropriate standard of care, and anesthesiologists could not successfully predict jury verdicts. The malpractice system appears to be operating far from its theoretical ideal if these results could be applied more generally. Thus, in practice, the legal system, which is to provide an optimal level of injury deterrence, may be a poor method to limit patient injury, improve patient safety, and provide compensation to negligently injured patients in the health delivery system. PMID- 10470628 TI - Preoperative pulse wave velocity fails to predict hemodynamic responses to anesthesia and to surgical stimulation. AB - STUDY OBJECTIVE: To determine whether pulse wave velocity (PWV), which is a useful indicator of cardiovascular disease, can predict the hemodynamic responses to anesthesia and surgical stimulation in surgical patients. DESIGN: Prospective, randomized study. SETTING: Operating rooms at a university hospital. PATIENTS: 30 ASA physical status I and II female patients presenting for elective gynecologic surgery. INTERVENTION: Patients who received one of the following anesthesia techniques were studied; isoflurane, sevoflurane and sevoflurane with nitrous oxide (N2O). MEASUREMENTS AND MAIN RESULTS: Patients underwent preoperative PWV measurements using an automatic PWV measurement device. Systolic blood pressure and heart rate levels were recorded at baseline, during anesthesia, and at skin incision. PWV had no significant correlation with percent changes of systolic blood pressure or heart rate, even after controlling the choice of anesthetic drugs. CONCLUSIONS: Preoperative PWV fails to predict hemodynamic responses to anesthesia and to surgical stimulation. PMID- 10470627 TI - Insertion of the cuffed oropharyngeal airway (COPA) with propofol or sevoflurane in adults. AB - STUDY OBJECTIVES: To compare the respiratory depressant effects of propofol and sevoflurane used to facilitate the placement of the cuffed oropharyngeal airway (COPA), and to evaluate the effectiveness of the COPA in supporting positive pressure ventilation during anesthetic-induced apnea. DESIGN: Randomized, single blinded study. SETTING: University hospital. PATIENTS: 60 ASA physical status I and II adult patients scheduled for elective surgery with general anesthesia. INTERVENTIONS: Patients were induced either with spontaneous inhalation of 5% sevoflurane or with propofol 2.0 mg/kg intravenously (i.v.) followed by a continuous infusion of 170 micrograms kg-1 min-1. If the propofol patient had a tight jaw in 90 seconds, additional propofol (0.5 mg/kg) was administered and the infusion rate was increased to 200 micrograms kg-1 min-1. The COPA was placed as soon as the jaw was sufficiently relaxed to allow its insertion into the mouth. MEASUREMENTS AND MAIN RESULTS: The median (range) time to the COPA placement were 90 seconds (30 to 150 sec) and 120 seconds (60 to 210 sec) with propofol and sevoflurane, respectively (p = 0.07, Mann-Whitney U-test). Unacceptable responses to the placement (3 or more coughs, vigorous or persistent [> 30 sec] movements) occurred in 23% and 17% of those who received propofol and sevoflurane, respectively, (p = 0.35. Chi-square test). All these responses were easily suppressed by additional doses of the assigned induction drug. After placement of the COPA, 53% (16/30) of the propofol patients had apnea lasting at least 30 seconds. Notably, the positive airway pressure at which a leak occurred in the mouth (pharyngeal leak pressure) was lower during this propofol-induced apnea than after the return of spontaneous breathing [9 (5 to 20) cmH2O vs. 15 (5 to 20) cmH2O, p < 0.01, Wilcoxon's signed-rank test]. In contrast, at no time were the sevoflurane patients apneic, and their pharyngeal leak pressure immediately following the placement was 12 (10 to 20) cmH2O. CONCLUSIONS: Propofol and sevoflurane are equally effective in facilitating the placement of the COPA. However, propofol often induces apnea, which is complicated by a less effective seal of the airway by the COPA against positive pressures. Because sevoflurane induction allows spontaneous respiration to continue and provides an adequate pharyngeal seal immediately following the placement of the COPA, it may be advantageous when apnea is not desired. PMID- 10470629 TI - Cerebral blood flow velocity during repeatedly induced ventricular fibrillation. AB - STUDY OBJECTIVE: To investigate the effect of induced ventricular fibrillation and defibrillation on cerebral blood flow (CBF) was investigated using a transcranial Doppler. DESIGN: Prospective clinical study. SETTING: University hospital. PATIENTS: 12 ASA physical status III and IV patients who underwent implantable cardioverter defibrillator placement during general anesthesia. INTERVENTIONS: Cerebral blood flow velocity was measured repeatedly during induced ventricular fibrillation and subsequent defibrillation. MEASUREMENTS AND MAIN RESULTS: The mean flow velocity in the middle cerebral artery was measured using a transcranial Doppler. The mean flow velocities decreased significantly immediately after ventricular fibrillation was induced, but they returned to preventricular fibrillation levels immediately after successful defibrillation. Repeatedly induced ventricular fibrillations have no cumulative detrimental effect on the CBF velocity. CONCLUSIONS: Repetitively induced ventricular fibrillation and defibrillation during the insertion of implantable cardioverter defibrillator did not show any detrimental changes in CBF. Transcranial Doppler may be a more sensitive device than other currently available cerebral monitors to detect changes in cerebral circulation during a brief episode of ventricular fibrillation and defibrillation. PMID- 10470630 TI - The optimal effective concentration of lidocaine to reduce pain on injection of propofol. AB - STUDY OBJECTIVE: To determine the optimal concentration of lidocaine that reduces pain on injection of a propofol-lidocaine mixture. DESIGN: Prospective, randomized, double-blinded, clinical investigation. SETTING: Medical center, university teaching hospital. PATIENTS: 240 ASA physical status I and II female outpatients, aged 21 to 65 years, undergoing dilation and curettage with propofol for anesthesia induction. INTERVENTIONS: Patients were randomized to one of four groups in double-blinded fashion. In Group A (control), patients were given propofol containing normal saline; in Group B, Group C, and Group D, patients received propofol containing 0.05% lidocaine (Group B), propofol containing 0.1% lidocaine (Group C), and propofol containing 0.2% lidocaine (Group D) for induction. MEASUREMENTS AND MAIN RESULTS: The incidence of pain on injection of propofol was significantly decreased in Group C and Group D (8.3% and 10.0%, respectively) in comparison to the control group (91.7%) (p < 0.001). Although the result in Group B (76.7%) was better than that in the control group, the difference was not statistically significant. No significant difference was seen between Group C and Group D. CONCLUSIONS: The optimal effective concentration of lidocaine, which decreased the incidence of pain caused by propofol injection, was 0.1% in the currently studied population. PMID- 10470631 TI - A collaborative regional ocular anesthesia training program: successes and failures. AB - STUDY OBJECTIVE: To describe a collaborative effort of the departments of ophthalmology and anesthesiology to teach anesthesiology residents regional ocular anesthesia; to detect any differences in positive or negative outcomes after blocks performed by anesthesiology residents versus blocks performed by ophthalmology residents. DESIGN: Prospective descriptive, study. SETTING: Outpatient surgery in a university-affiliated veterans affairs hospital. PATIENTS: 614 patients requiring elective ocular surgery. INTERVENTIONS: Outcomes from patients who underwent regional anesthesia performed by ophthalmology residents were compared to outcomes from patients who received anesthesia from anesthesiology residents. MEASUREMENTS AND MAIN RESULTS: A detailed description of the collaborative teaching program in ocular anesthesia is presented. Ophthalmology residents performed the majority of regional ocular blocks (87% vs. 13%). There was no statistical difference in the incidence of negative outcomes, such as retrobulbar hemorrhage, between ophthalmology residents and anesthesiology residents (3/534 vs. 1/80) or in the incidence of successful blocks (90% for ophthalmology residents vs. 88% for anesthesiology residents). CONCLUSIONS: Regional ocular anesthetic techniques can be safely and successfully taught to residents in anesthesiology. PMID- 10470632 TI - Xenon suppresses the hypnotic arousal in response to surgical stimulation. AB - STUDY OBJECTIVE: To evaluate the suppressive effects of xenon (Xe) on hypnotic arousal at skin incision. DESIGN: Prospective, randomized study. SETTING: Operating rooms at a university hospital. PATIENTS: 35 ASA physical status I and II patients presenting for elective lower abdominal surgery. INTERVENTIONS: Patients were randomly assigned to receive one of the following regimens: 1.3 minimum alveolar concentration (MAC) isoflurane, 1.3 MAC sevoflurane, 0.7 MAC Xe with 0.6 MAC sevoflurane, 1 MAC Xe with 0.3 MAC sevoflurane, or 0.7 MAC nitrous oxide (N2O) with 0.6 MAC sevoflurane (n = 7 each group). MEASUREMENTS AND MAIN RESULTS: The bispectral index (BIS) was measured at baseline, during anesthesia, and after skin incision. BIS increased significantly at skin incision from the values noted during anesthesia in the sevoflurane and N2O groups, whereas it remained stable at incision in the other three groups (mean change in BIS: 0 +/- 9 for isoflurane, 15 +/- 8 for sevoflurane, 5 +/- 6 for 0.7 MAC Xe, 4 +/- 11 for 1 MAC Xe, and 9 +/- 5 for N2O). CONCLUSIONS: Unlike N2O, Xe was able to suppress hypnotic arousal in response to surgical stimulation when administered with sevoflurane. PMID- 10470633 TI - Effects of tramadol and meperidine on respiration, plasma catecholamine concentrations, and hemodynamics. AB - STUDY OBJECTIVE: To evaluate the effects of high analgesic doses of tramadol and meperidine on respiration, plasma catecholamine concentrations, and hemodynamic parameters. STUDY DESIGN: Randomized, double-blind, cross-over, controlled volunteer study. SETTING: Laboratory at a university hospital. SUBJECTS: 8 healthy male volunteers. INTERVENTIONS: Tramadol was given as a 150 mg bolus plus a succeeding 3-hour steady infusion of 250 mg (83.3 mg/hr). Meperidine was given in a similar manner as a bolus dose of 112.5 mg plus 187.5 mg in a 3-hour steady infusion (62.5 mg/hr). Experimental pain was induced using a tourniquet. MEASUREMENTS AND MAIN RESULTS: Respiration was studied noninvasively with respiratory inductive plethysmography and pulse oximetry. Arterial line was used for measurement of hemodynamics and blood sampling. Tramadol did not have any clinically significant effects on respiration, breathing pattern, or hemodynamics, but an increase in plasma epinephrine levels was noted. Meperidine bolus decreased tidal volume (p < 0.05, difference from baseline) and pulse oxygen saturation (from 97% to 94%, p < 0.05), but during the succeeding infusion, the respiratory drive, measured as mean inspiratory flow, was enhanced (p < 0.05 difference from baseline), and the respiratory parameters returned to baseline level. No change in hemodynamics was noted, but a significant increase in plasma norepinephrine and epinephrine levels (from 0.9 to 1.6 nmol/L and from 0.3 to 0.8 nmol/L, respectively; p < 0.05) was observed after meperidine administration. Tramadol caused nausea more often than meperidine (p < 0.05, between treatments). CONCLUSIONS: Tramadol exhibited a minimal effect on respiration and breathing pattern in healthy volunteers. The respiratory effects of meperidine bolus were predictable with decreasing tidal volume and pulse oxygen saturation. In contrast, during meperidine infusion, adequate respiration was preserved despite the large amount of meperidine infused. PMID- 10470634 TI - Effect of peripheral perfusion on accuracy of pulse oximetry in children. AB - STUDY OBJECTIVE: To examine the effect of perfusion on accuracy of two pulse oximeters in children and to determine thresholds of perfusion below which these pulse oximeters become inaccurate or cease to function. DESIGN: Prospective, observational clinical study. SETTING: Operating room of a large university hospital. PATIENTS: 19 children 10 years of age or less, who were scheduled for general anesthesia with placement of an intraarterial catheter. INTERVENTIONS: A radial artery catheter, laser Doppler probe, skin temperature sensor, and band probes of two oximeters, Ohmeda 3700 (Boulder, CO) and Nellcor N200 (Hayward, CA), were attached to the same hand. Baseline pulse oximeter and Doppler readings were obtained with simultaneous hemoximetry (AVL Model 912 CO-Oxylite, Roswell, GA), skin and esophageal temperatures, total hemoglobin, and transduced arterial pressure. Readings of all parameters (n = 94) were obtained during periods of low perfusion or by occluding the upper arm to 70% to 100% of systolic pressure. MEASUREMENTS AND MAIN RESULTS: Bias (SpO2-SaO2) of each oximeter is compared to each perfusion variable (age, weight, core and skin temperature, hemoglobin concentration, pulse pressure, and percent flow by laser Doppler) to determine effect on accuracy. Data were analyzed using backward multivariate linear regression, Pearson correlation coefficients, and independent paired t-test. p < 0.05 was considered significant. Less than 2% bias is seen with either oximeter (Nellcor 1.55 +/- 2.33, Ohmeda 0.78 +/- 2.25). Independent predictors of bias for each machine include weight (r = -0.376; p < 0.001) and pulse pressure (r = 0.250; p = 0.021) for the Nellcor, and weight (r = -0.390; p < 0.001), percent flow by Doppler (r = 0.220; p = 0.035), and core temperature (r = 0.307; p = 0.003) for the Ohmeda. However, using predetermined thresholds for each variable, only skin temperature below 30 degrees C is identified as a significant predictor of oximeter inaccuracy. CONCLUSIONS: At the parameters explored in this study, the selected seven perfusion variables (age, weight, core and skin temperature, hemoglobin concentration, pulse pressure, and percent flow by laser Doppler) have little effect on accuracy of pulse oximetry in children. PMID- 10470635 TI - Effect of probe design on accuracy and reliability of pulse oximetry in pediatric patients. AB - STUDY OBJECTIVE: To determine if the traditional band-wrap disposable pulse oximeter probe is more accurate or reliable for oxygen monitoring in children than the reusable clip-type probe. DESIGN: Prospective, observational clinical study. SETTING: Operating room of a large university hospital. PATIENTS: 18 children 11 years of age or younger, who were scheduled for general anesthesia with placement of an intraarterial catheter. INTERVENTIONS: Pulse oximetry values were obtained using both band and clip probes of three pulse oximeters (Nellcor, Hayward CA; Novametrix, Wallingford, CT; Ohmeda, Boulder, CO) and compared with simultaneous hemoximetry values. Dropout rate (percent of down time) also was recorded for each probe-machine combination. MEASUREMENTS AND MAIN RESULTS: Data were analyzed using bias and precision and t-test. p < 0.05 is considered significant. Bias (mean SpO2-SaO2) is less than 2% for all probe-machine combinations. The range of error observed between SpO2 and SaO2 for all data points was greatest using the Nellcor band (27.8%) and least using the Ohmeda band (11.4%). In cyanotic children, the greatest bias and precision were observed with Nellcor band (N = 11, -5.12 +/- 9.74) and the best agreement with Novametrix band (N = 17, 0.08 +/- 4.21). The difference in bias for any test units above or below the median weight of 13.75 kg did not vary by more than 1%. Dropout rate was minimal for all units in nonbypass situations. After cardiopulmonary bypass, no data dropout was observed with Ohmeda band; observed down time with the other units varied between 34% and 55%. CONCLUSIONS: The type of probe selected has little effect on accuracy of pulse oximetry in children. After cardiopulmonary bypass, using the Ohmeda band combination may improve the likelihood of obtaining consistent readings and decreasing down time. PMID- 10470637 TI - Optimal length and angle of a new lightwand device (Trachlight). AB - STUDY OBJECTIVE: To investigate the effects of angle and length of the Trachlight lightwand and the effect of obesity on transillumination of the neck and difficulty of intubation. DESIGN: Prospective, randomized study. SETTING: Operating room of a university hospital. PATIENTS: 270 patients, aged 30 to 70 years, requiring anesthesia. INTERVENTION: Three different studies were performed. 1) One hundred twenty adults for general anesthesia to test the effect of the angle, 2) 120 patients for the length of the extrusion, and 3) 30 patients with body weight > or = 120% of the standard (Broca's formula) versus 30 with body weight < 120% of the standard to test the effect of the obesity. 1) The lightwand was bent at 20 degrees, 40 degrees, 60 degrees, or 80 degrees with 0 cm extruded from the endotracheal tube; 2) the lightwand was exposed from the endotracheal tube with -0.5, 0, 1, 2, 3, or 4 cm with 40 degrees angle; and 3) the effect of obesity was tested with 40 degrees angle and 0-cm extrusion. Endotracheal intubation was performed using the lightwand. Transillumination of the light at neck and the difficulty of intubation were assessed. MEASUREMENTS AND MAIN RESULTS: 1) A 20 degrees angle decreased the transillumination and an 80 degrees angle increased the number of difficult intubation. 2) Transillumination decreased with -0.5 cm and 0 cm extrusion. Difficult intubation increased with the extrusion > or = 3 cm. 3) Transillumination decreased and difficult intubation increased in the patients with body weight > or = 120% of the standard. CONCLUSIONS: To increase the success rate of tracheal intubation using the lightwand, the lightwand should be bent in 40 degrees to 60 degrees, with the extrusion of 1 to 2 cm from the tracheal tube. For obese patients with body weight > or = 120% of the standard, the lightwand was not useful. PMID- 10470636 TI - Effect of spinal versus general anesthesia on bladder compliance and intraabdominal pressure during transurethral procedures. AB - STUDY OBJECTIVE: To evaluate the influence of spinal versus general anesthesia on bladder compliance and intraabdominal pressure in elderly males undergoing elective transurethral resection of the prostate. DESIGN: Prospective, randomized, open-label study. SETTING: Teaching hospital. PATIENTS: 21 ASA physical status I, II, and III patients at least 18 years of age, undergoing transurethral surgery. INTERVENTIONS: According to a computer-generated randomization schedule, patients were allocated to one of two groups. In Group Spinal (S), 10 mg of hyperbaric tetracaine was administered intrathecally. In Group General Anesthesia (GA), patients received, fentanyl intravenous (i.v. 1 to 2 micrograms/kg and propofol i.v. 1.0 to 2.0 mg/kg for induction of anesthesia. Thereafter, a laryngeal mask airway was inserted and, with spontaneous ventilation, anesthesia was maintained by administering isoflurane (end-tidal 0.7% to 1.2%) and 70% nitrous oxide (N2O) in oxygen. Intraabdominal pressure and bladder compliance were recorded prior to the induction of anesthesia and immediately before the onset of the surgical procedure. MEASUREMENTS AND MAIN RESULTS: The two groups were demographically comparable. In Group S, mean bladder compliance was significantly (p = 0.003) higher and mean intraabdominal pressure significantly lower (p = 0.007) when compared to baseline preanesthetic values. In Group GA, mean intraabdominal pressure significantly (p = 0.006) decreased when compared to baseline preanesthetic recordings. Following the induction of general anesthesia, a small change in bladder compliance was noted. However, statistical significance was not reached. Data were analyzed and compared using Student's t-test (p < 0.05 was considered statistically significant). CONCLUSION: Both spinal and general anesthesia induced a significant decrease in intraabdominal pressure. While both techniques were associated with an increase in bladder compliance, statistical significance was demonstrated only in the spinal anesthesia treatment group. PMID- 10470638 TI - Intraoperative use of automated external defibrillator. AB - We report on the anesthetic management of a patient with peripartum cardiomyopathy and frequent episodes of ventricular tachycardia, who underwent surgery for tracheal stenosis. Prior to this surgery, the patient had been implanted with an automatic implantable cardioverter-defibrillator (AICD), placed abdominally. In the operating room, the AICD was deactivated, and an automated external defibrillator (AED) was placed. Intraoperatively, the AED identified and treated the patient's ventricular tachycardia. Advantages of the AED in this hospital setting included rapid response to the cardiomyopathy, safe, hands-free operation, and minimal disruption of the surgical procedure. Safety concerns when using the AED are also detailed. PMID- 10470639 TI - Anesthetic management of a patient with laryngeal amyloidosis. AB - A 73-year-old woman who suffered from progressive hoarseness for 6 years and dysphagia without pain for 1 year presented with a soft tissue deposition on the posterior region of the vocal cords and narrowing in the subglottic area. Biopsy of this soft tissue and histological examination revealed laryngeal amyloidosis. A tracheostomy and partial removal of the amyloid were performed with general anesthesia. The airway was secured with a smaller diameter endotracheal tube, which was inserted atraumatically with Magill's forceps. The larynx is a rare site for amyloidosis. Laryngeal amyloidosis is fragile and hemorrhagic. Therefore, massive bleeding may occur during intubation. Anesthetists should take care in intubating the tracheas of these patients and be aware of other systemic diseases in laryngeal amyloidosis. PMID- 10470640 TI - Anesthetic management of high-risk cardiac patients undergoing noncardiac surgery under the support of intraaortic balloon pump. AB - Patients with severely impaired left ventricular function, an uncorrectable coronary artery disease, and a recent myocardial infarction are at high risk of cardiac complications after major noncardiac surgery. We present two patients with extensive three-vessel coronary artery disease who underwent intraperitoneal surgery under the support of intraaortic balloon pump (IABP). In one patient, the IABP was inserted urgently because of the development of chest pain with significant ST depression on arrival in the operating room, and the other patient was managed with prophylactic IABP. There were no intraoperative or postoperative cardiac events in either patient. Thus, IABP should be considered in the perioperative management of patients with severe cardiac diseases. PMID- 10470641 TI - Use of the intubating laryngeal mask airway to facilitate awake orotracheal intubation in patients with cervical spine disorders. AB - Airway management in patients with unstable cervical spines remains a challenge for anesthesia providers. Because neurologic evaluations may be required following tracheal intubation and positioning for the surgical procedure, an awake intubation technique is desirable in this patient population. In this report, we describe the use of an intubating laryngeal mask airway (ILMA) to facilitate awake tracheal intubation in two patients with cervical spine disorders. After topical local analgesia, the ILMA was inserted easily, and a tracheal tube was passed through the glottic opening without complications. Thus, the ILMA may be an acceptable alternative to the fiberoptic bronchoscope for awake tracheal intubation. PMID- 10470642 TI - Efforts at intubation: cervical injury in an emergency circumstance? PMID- 10470643 TI - Pulse oximetry during and after angiography in infants and children. PMID- 10470644 TI - Asymmetry of cilia and of mice and men. AB - Evidence is given for the opinion that cilia in the early embryo, by their work, determine the laterality of the body; without ciliary work body laterality would be randomized. More exactly, monocilia in the primitive node are responsible for this determination. They have been described as being of the 9+0 type, but with dynein arms and with a gyrating movement. The orientation of the monocilia on the epithelium is of no importance but the direction of their gyration is, as may also be the shape of the node. The chirality of the cilia is thus reflected directly in the asymmetry of the body. The dynein arms go clockwise as seen from the base to tip and the ciliary rotation is in the same direction. The resulting waterflow is towards the left and so is the movement of the forming heart. In most subgroups of the immotile-cilia syndrome this mechanism does not work and equally many individuals will be born with situs inversus as with situs solitus. An exception is the immotile-cilia subgroup, named 'microtubule transposition', which is characterized by all cilia having a 9+0 structure throughout most of their length. PMID- 10470645 TI - Altered Hox expression and increased cell death distinguish Hypodactyly from Hoxa13 null mice. AB - Hypodactyly (Hoxa13Hd) mice have a small deletion within the coding sequence of Hoxa13 and a limb phenotype that is more severe than that of mice with an engineered null allele of Hoxa13. We used whole-mount in situ hybridization, Nile blue sulfate staining and genetic crosses to determine the basis for the phenotypic differences between these two mutants. Expression of Hoxd13 was unaffected in Hoxa13-/- mice, but its domain was reduced at the anterior and posterior margins of the autopod in Hoxa13Hd/Hd limb buds. The maturation of Hoxd11 expression was delayed and expression of Hoxa11 failed to become restricted to the autopod/zeugopod junction in both Hoxa13Hd/Hd and Hoxa13-/- limb buds compared to wild-type mice. Fgf8 expression was normal in both Hoxa13Hd/Hd and Hoxa13-/- mice throughout limb development. A dramatic increase in cell death was observed in limb bud mesenchyme of Hoxa13Hd/Hd mice as early as E11.5 but not in mice homozygous for the null allele. Genetic background was excluded as the basisforthe phenotypic differences. Compound heterozygotes (Hoxa13-/Hd) displayed an intermediate phenotype relative to both homozygotes suggesting that Hoxa13Hd has an effect on the development of the autopod beyond that which may result from a loss of HOXA13 protein. These results showthat Hoxa13Hd has a negative effect on the survival of the mesenchyme in the autopod, unlike the Hoxa13 null mutation, that cannot be explained by a failure of the AER to express Fgfs. In addition, at least one target of HOXA13 may be Hoxa11. PMID- 10470646 TI - Misexpression of the RNA-binding protein ELRB in Xenopus presumptive neurectoderm induces proliferation arrest and programmed cell death. AB - Proteins of the ELAV/Hu family share the presence of three RNA binding domains. In Xenopus, three nervous system-specific elav/Hu related genes, elrB, elrC and elrD, have been identified so far. The temporally regulated expression patterns of elrB, elrC and elrD suggest their involvement at different steps of neural differentiation. In the present study we misexpressed elrB by RNA injection in early Xenopus embryos and analyzed morphologically and molecularly its effects on neural development. We showed that heterochronous expression of elrB in presumptive neurectoderm down-regulates the expression of neural markers, such as N-tubulin, as well as that of other Xenopus elav-like genes, elrC and elrD, whereas ectopic expression of elrB in presumptive mesoderm has no effect on MyoD. Misexpression of elrB also induces severe defects in neural tube development, associated with massive cell loss resulting from early cell cycle arrest and programmed cell death. Our results are discussed in the context of early neural differentiation. PMID- 10470647 TI - The CNS midline cells and spitz class genes are required for proper patterning of Drosophila ventral neuroectoderm. AB - The Drosophila embryonic central nervous system (CNS) develops from sets of neuroblasts (NBs) which segregate from the ventral neuroectoderm during early embryogenesis. It is not well established how each individual NB in the neuroectoderm acquires its characteristic identity along the dorsal-ventral axis. Since it is known that CNS midline cells and spitz class genes (pointed, rhomboid, single-minded, spitz and Star) are required for the proper patterning of ventral CNS and epidermis originated from the ventral neuroectoderm, this study was carried out to determine the functional roles of the CNS midline cells and spitz class genes in the fate determination of ventral NBs and formation of mature neurons and their axon pathways. Several molecular markers for the identified NBs, neurons, and axon pathways were employed to examine marker gene expression profile, cell lineage and axon pathway formation in the spitz class mutants. This analysis showed that the CNS midline cells specified by single minded gene as well as spitz class genes are required for identity determination of a subset of ventral NBs and for formation of mature neurons and their axon pathways. This study suggests that the CNS midline cells and spitz class genes are necessary for proper patterning of the ventral neuroectoderm along the dorsal ventral axis. PMID- 10470648 TI - Cell lineage analysis of pattern formation in the Tubifex embryo. I. Segmentation in the mesoderm. AB - Annelids are strongly segmented animals that display a high degree of metamerism in their body plan. The embryonic origin of metameric segmentation was examined in an oligochaete annelid Tubifex using lineage tracers. Segmental organization arises sequentially in the anterior-to-posterior direction along the longitudinal axis of the mesodermal germ band, a coherent column of primary blast cells that are produced from the mesodermal teloblast. Shortly after its birth, each primary blast cell undergoes a spatiotemporally stereotyped sequence of cell divisions to generate three classes of cells (in terms of cell size), which together give rise to a distinct cell cluster. Each cluster is composed of descendants of a single primary blast cell; there is no intermingling of cells between adjacent clusters. Relatively small-sized cells in each cluster become localized at its periphery, and they form coelomic walls including an intersegmental septum to establish individuality of segments. A set of cell ablation experiments showed that these features of mesodermal segmentation are not affected by the absence of the overlying ectodermal germ band. These results suggest that each primary blast cell serves as a founder cell of each mesodermal segment and that the boundary between segments is determined autonomously. It is concluded that the metameric body plan of Tubifex arises from an initially simple organization (i.e., a linear series) of segmental founder cells. PMID- 10470649 TI - Evidence that platelet derived growth factor (PDGF) action is required for mesoderm patterning in early amphibian (Xenopus laevis) embryogenesis. AB - Mesoderm induction is one of the major events of early vertebrate embryonic patterning. It appears to be controlled by sequential and combinatorial actions of several kinds of peptide growth factors. These include activin, fibroblast growth factor (FGF), and transforming growth factor-beta (TGF-beta), among others. In the present study, the function of platelet-derived growth factor (PDGF) in early Xenopus laevis embryogenesis was investigated. In the animal-cap assay, PDGF caused pre-ectodermal tissue to develop a mesoderm specific morphology (elongation) and to express the mesoderm marker genes, MyoD family and alpha-cardiac actin. In addition, two other genes were expressed -related serum response factor SL1 (a dorsal mesodermal marker) and myosin light chain (MLC2 heart marker). A role for PDGF in normal (in vivo) mesoderm induction is implicated because injection of PDGF receptor alpha antisense RNA into 2-cell embryos erased the animal cap's mesoderm marker expression. Those injected embryos also exhibited morphological abnormalities including incomplete gastrulation, failure of neural fold closing, and abnormal somitogenesis. PMID- 10470650 TI - N-CAM is not required for initiation of secondary chondrogenesis: the role of N CAM in skeletal condensation and differentiation. AB - Condensation precedes chondrogenic differentiation during development of primary cartilage. While neural cell adhesion molecule (N-CAM) enhances condensation, it is unclear whether N-CAM is also required for initiation of chondrogenic differentiation. In this study, the role of N-CAM in secondary chondrogenesis from periosteal cells of the quadratojugal (QJ) from embryonic chicks was studied using several in vitro approaches. The QJ is a membrane bone and so is not preceded by cartilage formation during development. However, QJ periosteal cells can differentiate into chondrocytes to form secondary cartilage in vivo. When QJ periosteal cells were enzymatically released and plated in low density monolayer, clonal or agarose cultures, chondrogenesis was initiated in the absence of N-CAM expression. Furthermore, overexpression of the N-CAM gene in periosteal cells in monolayer culture significantly reduced the number of chondrocyte colonies, suggesting that N-CAM inhibits secondary chondrogenesis. In contrast, and consistent with expression in vivo, N-CAM is expressed during osteogenesis from QJ periosteal cells and mandibular mesenchyme in vitro. These results are discussed in relation to the role of N-CAM in osteogenesis and in primary and secondary condensation. PMID- 10470651 TI - IGF-I, IGF-II and insulin promote differentiation of spermatogonia to primary spermatocytes in organ culture of newt testes. AB - Recombinant human insulin-like growth factors (rhIGF-I and rhIGF-II) and human insulin promoted the differentiation of spermatogonia into primary spermatocytes in newt testes fragments cultured in a chemically defined medium. The biological potency for promoting differentiation was dose-dependent for all the ligands with the highest potency displayed by IGF-I, followed by IGF-II, and the least by insulin. The difference in potency was larger between IGF-II and insulin than that between IGF-I and IGF-II. This order of biological potency was in good accordance with the order of affinity in binding specificity of [125I]IGF-I to the testicular membrane fractions: IGF-II and insulin competed the binding of [125I]IGF-I only at concentrations 20-fold and 100-fold higher, respectively, than IGF-I. Specific binding was observed in both somatic cells (mostly Sertoli cells) and germ cells (spermatogonia and primary spermatocytes), though the binding to somatic cells was about 2.7 times higher than that to germ cells. These results indicate that (1) specific binding sites for IGF-I are present in the newt testes, (2) IGF-II and insulin also bind to these receptors but to a lesser degree, and (3) IGF-II and insulin as well as IGF-I promote spermatogonial differentiation into primary spermatocytes by binding to the IGF-I receptor. PMID- 10470652 TI - Characterization of stage-specific embryonic antigen-1 (SSEA-1) expression during early development of the turkey embryo. AB - SSEA-1 is a carbohydrate epitope associated with cell adhesion, migration and differentiation. In the present study, SSEA-1 expression was characterized during turkey embryogenesis with an emphasis on its role in primordial germ cell development. During hypoblast formation, SSEA-1 positive cells were identified in the blastocoel and hypoblast and later in the germinal crescent. Based on location and morphology, these cells were identified, as PGCs. Germ cells circulating through embryonic blood vessels were also SSEA-1 positive. During the active phase of migration, PGCs in the dorsal mesentery and gonad could no longer be identified using the SSEA-1 antibody. The presence of PGCs at corresponding stages was verified using periodic acid Schiff stain. Pretreatment of PGCs with trypsin, alpha-galactosidase and neuraminidase did not restore immunoreactivity to SSEA-1. In general, expression was not limited to the germ cell lineage. SSEA 1 was also detected on the ectoderm, yolk sac endoderm, gut and mesonephric tubules. During neural tube closure, SSEA-1 was expressed by the neural epithelium of the fusing neural folds. Later SSEA-1 was detected in regions of the developing spinal cord. Enzyme pretreatment unmasked the epitope on some neural crest cells and cells in the sympathetic ganglion. The temporal and spatial distribution of SSEA-1 in the turkey embryo suggests a role in early germ cell and neural cell development. The absence of SSEA-1 on turkey gonadal germ cells was different from that observed for the chick. Therefore, while features of avian germ cell development appear to be conserved, expression of SSEA-1 can vary with the species. PMID- 10470653 TI - Hoxb-5 is expressed in gill arch 5 during pharyngeal arch development of flounder Paralichthys olivaceus embryos. AB - Hox genes are expressed in domains with clear anterior borders exhibiting 3'-->5' hierarchy in hindbrain and in the pharyngeal area commonly in vertebrate embryos. Teleost embryos form seven pharyngeal arches, the mandibular arch, hyoid arch and the gill arches 1-5. We previously reported that, in Japanese flounder (Paralichthys olivaceus) embryos, Hoxd-4 is expressed from rhombomere 7 to the spinal cord in the central nervous system and at gill arches 2-5. At present, the hierarchy of Hox genes at gill arches 3-5 of teleost fish is unclear. Here, we investigated the expression domains of Hoxb-5 in the flounder embryo by whole mount in situ hybridization to gain insight into the Hox code at gill arches. The initial signal indicating Hoxb-5 expression was identified in the spinal cord at hatching, corresponding with the prim-5 stage of zebrafish. Then, intense signals were detected from the anterior part of the spinal cord and from the posterior part of the pharyngeal area at 36 h after hatching. By serially sectioning the hybridized embryos, it was found that signal in the pharyngeal area came from the most posterior gill arch 5. Therefore, it is speculated that Hoxb-5 functions in regional identification of gill arch 5 in this teleost. PMID- 10470654 TI - Amphibian embryos as a model system for organ engineering: in vitro induction and rescue of the heart anlage. AB - Beating hearts can be induced under in vitro conditions when the dorsal blastopore lip (including the zone of Spemann organizer) is treated with Suramin. In contrast, untreated organizer forms dorsal mesodermal derivatives as notochord and somites. When those in vitro produced heart precursor tissues are transplanted ectopically in the posterior trunk area of early larvae, secondary beating heart structures will be formed. Furthermore, the replacement of the heart primordium of the host embryo by heart tissue induced under in vitro conditions will result in the rescue of the heart anlage. This model could be a valuable tool for the study of the multi-step molecular mechanisms of heart structure induction under in vitro conditions and vasculogenesis after transplantation into the host embryo. PMID- 10470655 TI - Function of estrogen receptor (ER) in gene expression. PMID- 10470656 TI - Drug-induced apoptosis and p53, BCL-2 and BAX expression in breast cancer tissues in vivo and in fibroblast cells in vitro. AB - BACKGROUND: Chemotherapeutic management of breast cancers is a difficult task as they show significant differences in chemosensitivity. The present study was undertaken to determine the usefulness of the apoptosis-related factors as indicators of tumor sensitivity to 5'-deoxyfluorouridine (5'-DFUR) in breast cancers. METHODS: (1) Forty-six breast cancer patients were randomly assigned to a group in which oral 5'-DFUR (1200 mg/day) was administered for more than 5 days before operation (24 patients) and a control group who received no preoperative chemotherapy (22 patients). Surgical specimens were examined for the frequency of apoptotic cells [apoptotic index (AI)] by a terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling method and for the expression of p53, BCL-2 and BAX by immunohistochemical staining. (2) Normal human diploid fetal lung fibroblast, IMR90 and SV40 transformed IMR90 were exposed to 5-FU. Apoptotic cells were detected by flow cytometry and BCL-2 and BAX mRNAs by real-time quantitative RT-PCR analysis. RESULTS: (1) No significant difference in the AIs or in BCL-2 and BAX scores was observed between the 5'-DFUR treated and control groups. However, in the p53 negative subgroup (n = 36), AI and BAX scores were higher and BCL-2 scores lower in the 5'-DFUR group than in the control group (P = 0.006, 0.008 and 0.050, respectively). (2) The sensitivity of IMR90 was significantly decreased by SV40 transformation and the 5-FU-induced cytotoxicity was mainly due to induction of apoptosis. The BCL-2/BAX mRNA ratio was decreased in response to 5-FU in IMR90. These results correlated with our clinical data. CONCLUSIONS: Preoperative treatment with 5'-DFUR induced apoptosis and changes in BCL-2 and BAX expression in p53 negative breast cancers. p53 status, AI and the BCL-2/BAX ratio may be useful information for the choice of postoperative chemotherapy for breast cancer. PMID- 10470657 TI - Genetic instability did not lead to p53 mutations in an extremely early-onset breast cancer in a cancer-prone family. AB - BACKGROUND: A positive family history is a major contributor to risk of development of breast cancer. METHODS: Somatic and germ line mutations of p53 and genetic instability were evaluated for an extremely early-onset breast cancer case in a cancer-prone family. RESULTS: The mode of inheritance in this case was clearly autosomal dominant. DNA replication error was recognized by detecting microsatellite allelic alterations. However, mutations of p53 were not found at either somatic or germ-line level. CONCLUSION: These genetic studies suggest that an increased genetic instability did not lead to p53 gene mutations in this breast cancer patient. PMID- 10470658 TI - Serum c-erB-2 levels in monitoring of operable breast cancer patients. AB - BACKGROUND: Various methods and criteria are used to determine protein overexpression of c-erbB-2 and the clinical utility of c-erbB-2 is under investigation. We have reported previously that the levels of cytosol c-erbB-2 in breast cancer were significantly different between the clinical stages. METHODS: The levels of c-erbB-2 protein were determined in sera from 210 breast cancer patients using a sandwich enzyme immunoassay between November 1996 and March 1998. The cut-off level was set at 5.4 ng/ml for healthy female blood donors. RESULTS: First, serum c-erbB-2 levels were analyzed in 73 preoperative breast cancer patients with stage I-IIIB disease. The range and median values were 2.3 32.3 and 4.8 ng/ml, respectively. The positive rate was 38%. Overexpression of serum c-erbB-2 was significantly associated with tumor size, clinical stage, histological grade, lymphatic invasion, nodal status and overexpression of cytosol c-erbB-2, but not with hormonal receptor status and other clinico pathological factors. Second, c-erbB-2, CEA and CA15-3 in sera were examined in 157 postoperative breast cancer patients. In the 137 disease-free patients, specificities of c-erbB-2, CEA and CA15-3 were 72, 93 and 99%, respectively, but in the 20 first recurrent patients, these sensitivities were 80, 25 and 25%, respectively. CONCLUSIONS: These results suggest that serum c-erbB-2 protein is a useful marker for predicting aggressive behavior and first recurrence of breast cancer. PMID- 10470659 TI - Long-term follow-up results of adult patients with acute lymphocytic leukemia or lymphoblastic lymphoma treated with short-term, alternating non-cross-resistant chemotherapy: Japan Clinical Oncology Group Study 8702. Lymphoma Study Group. AB - BACKGROUND: Patients with acute lymphocytic leukemia (ALL) and those with lymphoblastic lymphoma (LBL) have overlapping clinical and immunophenotypic features and they have been treated with the same or very similar chemotherapy regimens. The goal of this multi-institutional phase II trial was to evaluate the therapeutic efficacy of a short-term, six-drug chemotherapy regimen for adult patients with untreated ALL or LBL. METHODS: Forty-six eligible patients, 41 with ALL and five with LBL, were treated with a short-term (planned total therapy duration; 36-38 weeks), simplified chemotherapy program; two courses of VEPA-L (vincristine, cyclophosphamide, prednisolone, doxorubicin, I-asparaginase plus intrathecal methotrexate and prednisolone) followed by four courses of M-VEPA (methotrexate plus VEPA), without the traditional maintenance therapy using daily 6-mercaptopurine and weekly methotrexate. RESULTS: Thirty-six (78%; 95% confidence interval 64-89%) of the 46 eligible patients achieved complete remission (CR). Among the 36 patients who achieved CR, four (11%) died of treatment complications, 26 (72%) relapsed and six (17%) remain alive in continuous CR. The median survival for all 46 eligible patients is 14 months and the median disease-free survival (DFS) for the 36 patients who achieved CR is 11 months. The estimate of the proportion of survival at 7 years of all 46 eligible patients is 15% at a median follow-up time of 96 months and that of DFS of the 36 patients achieving CR is 17% at a median follow-up time of 93 months. Subgroup analysis showed that an elevated serum C-reactive protein (CRP) level, age of 30 years or older, the presence of B-symptom and T-cell phenotype were likely to be associated with shortened survival. Although the observed CR rate (78%) is within the range of satisfaction, the long-term survival rate (15%) is inferior to those of published programs incorporating maintenance therapy. CONCLUSIONS: A fraction of adult patients with ALL or LBL are curable with a short-term, six-drug chemotherapy regimen. However, this simplified therapy of shorter duration cannot be recommended. PMID- 10470660 TI - Multiple primary cancers of the esophagus and thyroid gland. AB - The occurrence of multiple primary cancers in the aerodigestive tract is a well known phenomenon that has been explained by the concept of 'field carcinogenesis'. Metachronous or synchronous esophageal cancer has usually been identified in patients with head and neck cancer, gastric cancer or colon cancer. The incidence of multiple primary cancers of the esophagus and thyroid gland is very low. We treated four patients with synchronous cancers of the cervical esophagus and the thyroid gland. Histologically, all of the esophageal cancers were squamous cell carcinomas. Thyroid cancers were evaluated as papillary carcinoma or follicular carcinoma. Both the esophageal cancer and the thyroid cancer frequently metastasized to lymph nodes. All patients had multiple lymph nodes metastasis from the esophageal or the thyroid cancer. In one patient, both the esophageal and the thyroid cancers were detected in the same lymph node. Three of four patients died from recurrence of esophageal cancer. The prognosis of these patients was poor. In the treatment of esophageal carcinoma, cancers of other organs including the thyroid gland should be carefully investigated. PMID- 10470661 TI - Metachronous bile duct cancer in a patient surviving for a decade and undergoing curative surgery twice. AB - We report a 75-year-old woman with metachronous bile duct cancer who underwent curative resection twice and has survived for a decade. In 1989, she was admitted because her serum alkaline phosphatase level was elevated. Computed tomography (CT) showed a low-density mass, 2 cm in diameter, at the left hepatic duct and intrahepatic bile duct dilatation in the left lobe. We diagnosed the lesion as an intrahepatic bile duct cancer and performed extended left hepatic lobectomy with systematic lymph node dissection. The histological diagnosis was a well differentiated cholangiocellular carcinoma with hepatic hilar and celiac lymph node metastases (T1N2M0, Stage IVB). In 1996, she was re-admitted with obstructive jaundice. CT showed a slightly enhanced mass, 4 cm in diameter, in the pancreatic head. After reducing the jaundice by percutaneous transhepatic biliary drainage, pancreatoduodenectomy was performed. The histological diagnosis of this lesion was a moderately differentiated adenocarcinoma originating from the intrapancreatic bile duct. Ten years after the first operation, she is leading a normal daily life with no cancer recurrence. These findings suggest that repeated curative surgery can result in a long-term survival of patients with metachronous bile duct cancer. PMID- 10470662 TI - Large bursa formation associated with osteochondroma of the scapula: a case report and review of the literature. AB - Bursitis or large bursa formation associated with osteochondroma has rarely been reported. A 33-year-old male presented with upper back pain, a rapidly developing mass beside the lateral border of his right scapula and snapping elicited by movement of the scapula. Plain radiograms and CT revealed osteochondroma on the ventral surface of the scapula without any unmineralized component and a huge cystic lesion around the osteochondroma. Aspiration of the cystic lesion showed the presence of sero-sanguineous fluid. MRI following the aspiration showed a thin cartilaginous cap with distinct outer margin and no soft tissue mass around the cap. Pathological examinations confirmed the diagnosis of osteochondroma with the large bursa formation. Clinical examination 19 months postoperatively showed an uneventful clinical course. PMID- 10470663 TI - Cancer incidence and incidence rates in Japan in 1994: estimates based on data from seven population-based cancer registries. The Research Group for Population based Cancer Registration in Japan. PMID- 10470664 TI - Serum prostate specific antigen levels are not elevated in most of the patients with advanced pancreatic adenocarcinoma. PMID- 10470665 TI - Knowing your patients. PMID- 10470667 TI - The thrill of victory, or the agony of defeat? PMID- 10470666 TI - Childhood rhabdomyosarcoma of the trunk and extremities. AB - Since 1979, 15 children with rhabdomyosarcoma have been treated at our institution. Included in this group are six children who presented with rhabdomyosarcoma of an extremity or trunk, requiring the use of combined multimodality therapy. The patients were clinically grouped and treated in accordance with the Intergroup Rhabdomyosarcoma Study protocol. All patients received combination chemotherapy based on their respective stage of disease at diagnosis. When feasible, the primary tumor was resected en bloc before chemotherapy was begun. After surgery, patients with unclear postoperative surgical margins and an initial good response to chemotherapy received radiotherapy to the primary site and at the regional lymphatics. Three of six patients developed or maintained a complete tumor response to induction chemotherapy. Radiotherapy maintained control of local disease in both groups. Overall, four patients, including one with disseminated disease at diagnosis, are alive, with a median survival time from diagnosis of 39 months. In children, treatment must be individualized, but complete local excision of the tumor with a tumor-free margin should be the goal. Major ablative amputation surgery was not performed. PMID- 10470669 TI - Fixing intertrochanteric fractures. PMID- 10470668 TI - The effects of single-axis balance board intervention on balance parameters in the elderly. AB - Falls among the elderly contribute in large part to the morbidity and mortality statistics of this age group. To date, few institutionalized programs exist to enhance balance among the elderly as a fall prevention adjunct. A 30-day balance program was instituted at independent living facilities in our community, and the effects were assessed for subjective and objective improvement in balance parameters. The results indicated that 97% of the participants enjoyed the program and found it to be nonstressful. Subjective improvement in confidence level and a diminished sense of balance loss was seen in 95% of the subjects. Significant improvement was noted in all tests measuring balance parameters. Objective improvement over baseline levels was seen in 90% of the study group. PMID- 10470670 TI - Orthopedic patients' satisfaction with their health care plan: a study performed by the Research and Education Committee of the Texas Orthopaedic Association. AB - To determine the satisfaction of orthopedic patients in regards to their health care plan, a four-page questionnaire was designed, pilot tested, and sent to 200 orthopedic surgeons randomly selected from members of the American Academy of Orthopaedic Surgeons in Texas. Fifty-seven orthopedic surgeons were not eligible to participate; our eligible cohort was 143 orthopedic surgeons. These surgeons were asked to administer the questionnaire to 20 randomly selected patients. Fifty-six of the 143 orthopedic surgeons participated, and surveys were completed by 939 patients. Patients were asked to rate their satisfaction with their health insurance, access to primary care physicians, and access to orthopedic surgeons. Although satisfaction was generally high, the patients with a gate-keeper (no direct access to specialists) were significantly less satisfied with their access to primary care physicians and orthopedic surgeons than those without a gatekeeper. PMID- 10470671 TI - Ganglions of the proximal interphalangeal joint. AB - Ganglion cysts-the most common hand tumors-usually affect women in their twenties and thirties. The cause of these cysts is unknown, although trauma has been postulated as an inciting factor. Ganglions occur most commonly at the dorsal and palmar wrist. However, ganglions of the proximal interphalangeal (PIP) joint are rare. Four patients with PIP joint ganglions were recently treated at our institution. Three received aspiration and one received operative therapy, all with good results. All four patients were older than 65 years. PMID- 10470672 TI - Superior mesenteric artery syndrome after segmental instrumentation: a biomechanical analysis. AB - Superior mesenteric artery (SMA) syndrome is a rare complication that occurs after correction of scoliosis. A review of the literature suggests that the incidence of this syndrome may be decreasing, as methods for the correction of scoliosis have improved. We present the first report of SMA syndrome that occurred after segmental 'derotational' instrumentation. Computerized axial tomography was helpful in suggesting the diagnosis of this condition. The risk factors and pathophysiology of SMA syndrome are reviewed with respect to the biomechanical effects of casting and Harrington and segmental instrumentation systems on the spine. Attention to the three-dimensional nature of the scoliotic deformity, particularly in the sagittal plane, may help to predict those at risk for the development of this potentially fatal syndrome. PMID- 10470673 TI - Low-energy scapular body fracture: a case report. AB - Scapula fractures are relatively rare and most often the result of high-energy trauma. However, they should always be included in a physician's differential diagnosis when a patient has a complaint of shoulder pain after trauma or violent muscular contraction about the shoulder. Because the vast majority are the result of a high-energy mechanism of injury, the physician should, as always, completely evaluate the patient for associated injuries. Most scapula fractures can be diagnosed on physical examination with localized tenderness, swelling, and hematoma formation over the fracture site. Radiographic confirmation and evaluation is routinely made using the three-view trauma series of the shoulder; additional views are rarely indicated. Treatment, consisting of a sling or sling and swath for comfort, mild narcotic medication, and early range-of-motion exercises virtually always leads to union and good glenohumeral function. Operative treatment is rarely indicated. A case of an low-energy isolated scapular body fracture sustained by a 41-year-old man is presented. PMID- 10470674 TI - A 33-year-old woman with an abdominal mass and multiple bone lesions. PMID- 10470675 TI - Scyphostatin, a neutral sphingomyelinase inhibitor from a discomycete, Trichopeziza mollissima: taxonomy of the producing organism, fermentation, isolation, and physico-chemical properties. AB - We performed experiments to screen for neutral sphingomyelinase inhibitors using rat brain microsomes as an enzyme source. Among more than 10,000 microbial extracts tested, a mycelial extract of Trichopeziza mollissima SANK 13892 exhibited potent inhibitory activity. The active compound, scyphostatin, was purified by a series of chromatographies. Scyphostatin inhibited the enzyme with an IC50 value of 1.0 microM. PMID- 10470676 TI - Biological activities of scyphostatin, a neutral sphingomyelinase inhibitor from a discomycete, Trichopeziza mollissima. AB - Scyphostatin is a specific inhibitor for mammalian neutral magnesium-dependent sphingomyelinase with a fifty percent inhibition concentration (IC50) value of 1.0 microM. When used to inhibit lysosomal acid sphingomyelinase, an approximately 50-fold greater concentration is required. In human peripheral monocytes, the compound inhibits bacterial lipopolysaccharide (LPS)-induced prostaglandin E2 production and LPS-induced interleukin-1beta production with IC50 values of 0.8 microM and 0.1 microM, respectively. In rat, p.o. administration of the compound has also been shown to inhibit carrageenin-induced paw edema. Thus, it is hoped that utility of scyphostatin as a pharmacological tool will contribute to our understanding of the role of ceramide in the cellular inflammation process. PMID- 10470677 TI - WF14861, a new cathepsins B and L inhibitor produced by Colletotrichum sp. I. Taxonomy, production, purification and structure elucidation. AB - WF14861, a novel cathepsins B and L inhibitor, was obtained from the culture mycelium of a fungus strain Colletotrichum sp. No. 14861. Spectroscopic analysis showed that WF14861 consisted of trans-epoxysuccinic acid, L-tyrosine and spermidine, WF14861 inhibited cathepsins B and L selectively. PMID- 10470678 TI - WF14861, a new cathepsins B and L inhibitor produced by Colletotrichum sp. II. Biological properties. AB - WF14861, 3-(N-(1-(N-(4-aminobutyl)-N-(3-aminopropyl)carbamoyl)-2-(4-hydroxyphenyl )ethyl)carbamoyl)oxirane-2-carboxylic acid, was obtained from the culture mycelium of Colletotrichum sp. as a novel cathepsins B and L inhibitor. WF14861 also showed inhibitory activities against bone derived crude protease and other cysteine proteases in vitro. The compound ameliorated the tissue damage and the bone destruction models of low-calcium-diet-fed mouse and adjuvant arthritis rat model. PMID- 10470679 TI - Conagenin derived from Streptomyces roseosporus enhances macrophage functions. AB - In contrast to the studies that describes the effects of conagenin (CNG) on the cellular immunity of lymphocytes (references), we investigated the in vitro effect of CNG on macrophage function. Phagocytosis of alveolar macrophages (AM) against opsonized-sheep red blood cells (SRBC) was significantly enhanced following in vitro incubation with CNG for 12 hours at 37 degrees C, which was closely associated with increased expression of Fc-receptor in AM membranes. Macrophage-activating factor (MAF), prepared from splenic lymphocytes in vitro stimulated with concanavarin A (Con A) for 48 hours at 37 degrees C, had also the enhancing effect on phagocytosis of AM against opsonized-SRBC. Preincubation with CNG (0.1 microg/ml) and MAF (1/100 dilution) induced the additional effect on phagocytosis of AM, which was associated with the increased expression of Fc receptor in AM membranes. These results suggest that CNG enhances AM phagocytosis by increasing the expression of Fc-receptor on their membranes via either effecting different sub-populations of AM cells or by activating independent mechanism on the same AM cell population. PMID- 10470680 TI - Efficacy of antibacterial drugs in mice with complex infection by Candida albicans and Escherichia coli. AB - We investigated the effect of seven antibacterial antibiotics: kanamycin, gentamicin, tetracycline, minocycline, ampicillin, piperacillin and cefotaxime, on survival of mice infected sequentially with a lethal dose of Candida albicans and a sublethal dose of Escherichia coli. The mortality of C. albicans-infected mice was facilitated by the superinfection with E. coli. When administered to mice with C. albicans/E. coli complex infection, aminoglycosides and tetracyclines significantly prolonged the survival period as compared with the infected and untreated controls. The recovery of viable counts of E. coli from the renal tissues was rapidly reduced by the treatment with gentamicin or minocycline, compared to the untreated control. Thus it was concluded that nullification by the treatment with aminoglycosides or tetracyclines of the enhancing effect of E. coli superinfection on the lethality of C. albicans infected mice is due to early elimination of E. coli from the kidney. PMID- 10470681 TI - Molecular cloning of the gene for the key carbocycle-forming enzyme in the biosynthesis of 2-deoxystreptamine-containing aminocyclitol antibiotics and its comparison with dehydroquinate synthase. AB - The 2-deoxystreptamine aglycon is a common structural feature found in aminocyclitol antibiotics including neomycin, kanamycin, tobramycin, gentamicin, sisomicin, butirosin and ribostamycin. A key enzyme involved in the biosynthesis of the 2-deoxystreptamine moiety is 2-deoxy-scyllo-inosose (DOI) synthase which catalyses the carbocycle formation from D-glucose-6-phosphate to 2-deoxy-scyllo inosose. The recent success of isolating the 2-deoxy-scyllo-inosose synthase from Bacillus circulans prompted us to clone the gene responsible for this important enzyme by the use of reverse genetics approach. With the aid of DNA probes constructed on the basis of the amino-terminal sequence of the purified 42 kDa subunit of the enzyme, the responsible gene btrC was successfully cloned. Subsequently the btrC gene was heterologously expressed in Escherichia coli, and the 2-deoxy-scyllo-inosose synthase activity of the recombinant polypeptide was confirmed by chemical analysis. The btrC gene encodes a protein composed of 368 amino acids with a molecular mass of 40.7 kDa. Our previous proposal for the similarity of 2-deoxy-scyllo-inosose synthase to dehydroquinate synthase has been confirmed on the basis of their amino acid sequences. Significant differences in the sequences can also be observed however, particularly in the crucial substrate recognition regions. Comparison of the BtrC sequence with those of biosynthetic enzymes for other related microbial products is also discussed. PMID- 10470682 TI - Alutenusin, a specific neutral sphingomyelinase inhibitor, produced by Penicillium sp. FO-7436. PMID- 10470683 TI - TMC-169, a new antibiotic of the aspochalasin group produced by Aspergillus flavipes. PMID- 10470684 TI - Cinnatriacetins A and B, new antibacterial triacetylene derivatives from the fruiting bodies of Fistulina hepatica. PMID- 10470685 TI - Novel nikkomycins Lx and Lz produced by genetically engineered Streptomyces tendae Tu901. PMID- 10470686 TI - Roselipins, novel fungal metabolites having a highly methylated fatty acid modified with a mannose and an arabinitol. PMID- 10470687 TI - Mer-f3, 12-hydroxy-ovalicin, produced by Metarrhizium sp. f3. PMID- 10470688 TI - Guns, lies, and videotape. PMID- 10470689 TI - Is neoadjuvant chemotherapy the answer for bladder cancer? PMID- 10470690 TI - What link between early respiratory viral infections and atopic asthma? PMID- 10470691 TI - Clinical value of aortic pulse-wave velocity measurement. PMID- 10470692 TI - Maternal age-standardisation of prevalence of Down's syndrome. PMID- 10470693 TI - Cause of adjacent-segment disease after spinal fusion. PMID- 10470694 TI - Sleep-disordered breathing in chronic stable heart failure. PMID- 10470695 TI - Population density and risk of childhood acute lymphoblastic leukaemia. PMID- 10470696 TI - Neoadjuvant cisplatin, methotrexate, and vinblastine chemotherapy for muscle invasive bladder cancer: a randomised controlled trial. International collaboration of trialists. AB - BACKGROUND: Several non-randomised trials have shown that transitional-cell carcinoma of the bladder is a moderately chemosensitive tumour. We investigated whether the addition of neoadjuvant cisplatin-based chemotherapy to radical surgery or radiotherapy would improve survival. METHODS: Patients with T2 G3, T3, T4a, N0-NX, or M0 transitional-cell carcinoma of the bladder undergoing curative cystectomy or full-dose external-beam radiotherapy were randomly assigned three cycles of neoadjuvant chemotherapy (cisplatin, methotrexate, and vinblastine, with folinic acid rescue, n=491) or no chemotherapy (n=485). When possible, clinical tumour response was assessed cytoscopically after completion of chemotherapy but before cystectomy or radiotherapy; histopathologically assessed response was on cystectomy samples. We recorded every 6 months locoregional persistence or relapse of tumour, appearance of distant metastases, survival, and cause of death. FINDINGS: Median follow-up of patients still alive was 4.0 years. 485 patients died, and 78.6% of deaths were due to transitional-cell carcinoma. Chemotherapy mortality was 1% and operative (cystectomy) mortality was 3.7%. Kaplan-Meier curves compared by means of the log-rank test gave a calculated absolute difference between groups in 3-year survival of 5.5% (95% CI -0.5 to 11.0, p=0.075; 55.5% for chemotherapy, 50.0% for no chemotherapy). Median survival in the chemotherapy group was 44 months compared with 37.5 months for the no-chemotherapy group. 32.5% of cystectomy samples contained no tumour after neoadjuvant chemotherapy. INTERPRETATION: Three cycles of neoadjuvant chemotherapy before cystectomy or radiotherapy did not give the 10% improvement in 3-year survival that was judged to be necessary for introduction into routine use. The chemotherapy regimen was associated with a higher pathological complete response rate in primary tumours, but there was no clear evidence that it would increase survival. PMID- 10470697 TI - Respiratory syncytial virus in early life and risk of wheeze and allergy by age 13 years. AB - BACKGROUND: The relation between lower respiratory tract illnesses in early life caused by the respiratory syncytial virus (RSV) and the subsequent development of wheezing and atopy in childhood is not well understood. We studied this relation in children who had lower respiratory tract illnesses that occurred before 3 years of age. METHODS: Children were enrolled at birth and cases of lower respiratory tract illness were ascertained by a physician. Viral tests were done for specimens collected at the time of the illness. Children were classified into five groups according to type and cause of lower respiratory tract illness. Children were then followed prospectively up to age 13, and we measured frequency of wheezing, pulmonary function, and atopic status (allergy skin-prick tests, serum IgE concentrations). FINDINGS: RSV lower respiratory tract illnesses were associated with an increased risk of infrequent wheeze (odds ratio 3.2 [95% CI 2.0-5.0], p < 0.001), and an increased risk of frequent wheeze (4.3 [2.2-8.7], p < or = 0.001) by age 6. Risk decreased markedly with age and was not significant by age 13. There was no association between RSV lower respiratory tract illnesses and subsequent atopic status. RSV lower respiratory tract illnesses were associated with significantly lower measurements of forced expiratory volume (2.11 [2.05-2.15], p < or = 0.001) when compared with those of children with no lower respiratory tract illnesses, but there was no difference in forced expiratory volume after inhalation of salbutamol. INTERPRETATION: RSV lower respiratory tract illnesses in early childhood are an independent risk factor for the subsequent development of wheezing up to age 11 years but not at age 13. This association is not caused by an increased risk of allergic sensitisation. PMID- 10470698 TI - Effects of Plasmodium vivax malaria in pregnancy. AB - BACKGROUND: Plasmodium vivax is more common than P. falciparum as a cause of malaria in many parts of the tropics outside Africa. P. falciparum infection has harmful effects in pregnancy, but the effects of P. vivax have not been characterised. We investigated the effects of P. vivax infection during pregnancy. METHODS: Since 1986, pregnant Karen women living in camps for displaced people on the western border of Thailand have been encouraged to attend antenatal clinics. Karen women were screened for malaria and anaemia at each week of pregnancy until delivery, and pregnancy outcome recorded. We compared the effects of P. vivax infection on anaemia and pregnancy outcome with those of P. falciparum and no malaria infection in the first pregnancy recorded at the antenatal clinics. FINDINGS: There were 634 first episodes of pure P. vivax malaria in 9956 women. P. vivax malaria was more common in primigravidae than in multigravidae and was associated with mild anaemia and an increased risk of low birthweight (odds ratio 1.64 [95% CI 1.29-2.08], p<0.001). The birthweight was a mean of 107 g (95% CI 61-154) lower in women with P. vivax infection than in uninfected women. By contrast with P. falciparum malaria, the decrease in birthweight was greater in multigravidae. P. vivax malaria was not associated with miscarriage, stillbirth, or with a shortened duration of pregnancy. INTERPRETATION: P. vivax malaria during pregnancy is associated with maternal anaemia and low birthweight. The effects of P. vivax infection are less striking than those of P. falciparum infection, but antimalarial prophylaxis against P. vivax in pregnancy may be justified. PMID- 10470699 TI - Activation-induced T-cell death and immune dysfunction after implantation of left ventricular assist device. AB - BACKGROUND: Cardiac transplantation is a limited option for end-stage heart failure because of the shortage of donor organs. Left-ventricular assist devices (LVADs) are currently under investigation as permanent therapy for end-stage heart failure, but long-term successful device implantation is limited because of a high rate of serious infections. To examine the relation between LVAD-related infection and host immunity, we investigated immune responses in LVAD recipients. METHODS: We compared the rate of candidal infection in 78 patients with New York Heart Association class IV heart failure who received either an LVAD (n=40) or medical management (controls, n=38). Fluorochrome-labelled monoclonal antibodies were used in analyses of T-cell phenotype. Analysis of T-cell function included intradermal responses to recall antigens and proliferative responses after stimulation by phytohaemagglutinin, monoclonal antibodies to CD3, and mixed lymphocyte culture. We measured T-cell apoptosis in vivo by annexin V binding, and confirmed the result by assessment of DNA fragmentation. Activation-induced T cell death was measured after T-cell stimulation with antibodies to CD3. All immunological tests were done at least 1 month after LVAD implantation. Between group comparisons were by Kaplan-Meier actuarial analysis and Student's t test. FINDINGS: By 3 months after implantation of LVAD, the risk of developing candidal infection was 28% in LVAD recipients, compared with 3% in controls (p=0.003). LVAD recipients had cutaneous anergy to recall antigens and lower (<70%) T-cell proliferative responses than controls after activation via the T-cell receptor complex (p<0.001). T cells from LVAD recipients had higher surface expression of CD95 (Fas) (p<0.001) and a higher rate of spontaneous apoptosis (p<0.001) than controls. Moreover, after stimulation with antibodies to CD3, CD4 T-cell death increased by 3.2-fold in LVAD recipients compared with only 1.2-fold in controls (p<0.05). INTERPRETATION: LVAD implantation results in an aberrant state of T cell activation, heightened susceptibility of CD4 T cells to activation-induced cell death, progressive defects in cellular immunity, and increased risk of opportunistic infection. PMID- 10470700 TI - 4G/5G promoter polymorphism in the plasminogen-activator-inhibitor-1 gene and outcome of meningococcal disease. Meningococcal Research Group. AB - BACKGROUND: Intravascular coagulation with infarction of skin, digits, and limbs is a characteristic feature of meningococcal sepsis. Children with meningococcal sepsis have higher than normal concentrations of plasminogen activator inhibitor 1 (PAI-1) in plasma. Combined with the widespread venous thrombosis, this finding suggests an impairment of fibrinolysis. A common functional insertion/deletion (4G/5G) polymorphism exists in the promoter region of the PAI-1 gene. We tested the hypothesis that children with the 4G/4G genotype produce higher concentrations of PAI-1, develop more severe coagulopathy, and are at greater risk of death during meningococcal sepsis. METHODS: The relation between meningococcal disease outcome, PAI-1 concentration, and PAI-1 genotype was investigated in 175 children with meningococcal disease (37 from Rotterdam, the Netherlands, and 138 from London, UK) and 226 controls (137 from Rotterdam, 89 from London). PAI-1 concentrations in plasma were measured by ELISA, and the 4G/5G PAI-1 polymorphism was detected by PCR and hybridisation. FINDINGS: Concentrations of PAI-1 on admission correlated with presentation (sepsis or meningitis) and outcome. The median PAI-1 concentration in children who died was substantially higher than that in survivors (2448 [IQR 1115-3191] vs 370 [146 914] ng/mL; p<0.0001). Patients with the 4G/4G genotype had significantly higher PAI-1 concentrations than those with the 4G/5G or 5G/5G genotype (1051 [550-2440] vs 436 [198-1225] ng/mL; p=0.03), and had an increased risk of death (relative risk 2.0 [1.0-3.8] for the two cohorts combined, and 4.8 [1.8-13] for the London cohort). INTERPRETATION: A genetic predisposition to produce high concentrations of PAI-1 is associated with poor outcome of meningococcal sepsis. This finding suggests that impaired fibrinolysis is an important factor in the pathophysiology of meningococcal sepsis. PMID- 10470701 TI - Variation in plasminogen-activator-inhibitor-1 gene and risk of meningococcal septic shock. AB - BACKGROUND: Some patients infected with Neisseria meningitidis develop septic shock accompanied by fibrin deposition and microthrombus formation in various organs, whereas others develop bacteraemia or meningitis without septic shock. We investigated whether genetic differences in the fibrinolytic system influence the development of meningococcal septic shock. METHODS: We investigated 50 patients who survived meningococcal infection, and 131 controls from the same geographical region. Because we had no information on genotypes of patients who died, we also genotyped 183 first-degree relatives of a consecutive series of patients with meningococcal infection for the 4G/5G deletion/insertion polymorphism in the promoter region of the plasminogen-activator-inhibitor-1 gene (PAI-1). The 4G allele is associated with increased gene transcription in cell lines in vitro and with increased PAI-1 concentrations in carriers in vivo. FINDINGS: The allele frequencies of 4G and 5G were similar between patients and controls. However, the 4G/4G genotype was present in only 9% of relatives of patients with meningitis compared with 36% of relatives of patients with septic shock. The 5G/5G genotype was more common among relatives of patients with meningitis (31 vs 11%, p=0.001). Patients whose relatives were carriers of the 4G/4G genotype had a six-fold higher risk of developing septic shock than meningitis (odds ratio 5.9 [95% CI 1.9-18.0]) compared with all other genotypes. INTERPRETATION: Variation in the PAI-1 gene does not affect the probability of contracting meningococcal infection, but does influence the development of septic shock. PMID- 10470702 TI - Stopping the pills. PMID- 10470703 TI - Tetanus and pregnancy-related mortality in Bangladesh. AB - A national survey of health-care providers in Bangladesh identified 298 women who died from pregnancy-related tetanus. Immunising all girls with tetanus toxoid and providing safe menstrual regulation services would prevent such deaths. PMID- 10470704 TI - Checklist for ruling out pregnancy among family-planning clients in primary care. AB - Where pregnancy tests are unavailable, health providers, fearing possible harm to fetuses, often deny contraception to nonmenstruating clients. In Kenya, a trial of a simple checklist to exclude pregnancy showed a good negative predictive value, which could improve access to service and reduce unwanted pregnancies and their sequelae. PMID- 10470705 TI - Olanzapine-induced agranulocytosis. AB - The antipsychotic drug olanzapine was developed to reduce the risk of haematotoxicity. We report a case of agranulocytosis induced by olanzapine. PMID- 10470706 TI - Olanzapine-induced neutropenia after clozapine-induced neutropenia. AB - Olanzapine is a new antipsychotic drug, pharmacodynamically similar to clozapine, but putatively devoid of haematological iatrogenicity. We report a case of relapse of previously clozapine-induced neutropenia after olanzapine treatment. PMID- 10470707 TI - Tryptophan hydroxylase autoantibodies and intestinal disease in autoimmune polyendocrine syndrome type 1. AB - Tryptophan hydroxylase antibodies, associated with gastrointestinal disease in autoimmune polyendocrine syndrome type 1, are specific for this disease and not present in patients with other bowel disorders or healthy controls. PMID- 10470708 TI - Intravenous immunoglobulin compared with cyclophosphamide for proliferative lupus nephritis. AB - Among 14 randomised patients with proliferative lupus nephritis, monthly intravenous immunoglobulin maintained remission over 18 months, similar to standard intravenous cyclophosphamide treatment. Pulsed immunoglobulin may be a useful alternative therapy in lupus nephritis. PMID- 10470709 TI - Therapeutic implications of the sentinel lymph node in breast cancer. AB - Immunohistochemistry of histologically negative axillary lymph nodes in breast cancer patients resulted in upstaging of the sentinel lymph node in eight (14%) of 52 patients. The resulting information altered clinical management in six of these patients. Thus, this technique may affect clinical decision-making in breast-cancer patients. PMID- 10470710 TI - Pad urine collection for early childhood urinary-tract infection. AB - Urine collected into absorbent pads is commonly contaminated, making this method unreliable for ruling out urinary-tract infection in young children. Bag samples are equally unhelpful, whereas clean-catch samples are least likely to need repeating. PMID- 10470711 TI - Patients' perceptions of intensive care. AB - Pain, noise, sleep deprivation, thirst, hunger, heat, cold, fear, anxiety, isolation, physical restraint, want of information, and absence of daylight were common memories of patients surviving intensive care. PMID- 10470712 TI - A human bite. AB - We report the transmission of group A streptococci by a human bite leading to severe necrotising fasciitis. Rapid surgical and antibiotic treatment led to healing without fractional loss of the patient's infected leg. PMID- 10470713 TI - Artificial implants: making the marriage work. PMID- 10470714 TI - Another step towards reshaping peer review at the NIH. PMID- 10470715 TI - Patients recalled after endoscope contamination. PMID- 10470716 TI - Epidemiological features of chronic low-back pain. AB - Although the literature is filled with information about the prevalence and incidence of back pain in general, there is less information about chronic back pain, partly because of a lack of agreement about definition. Chronic back pain is sometimes defined as back pain that lasts for longer than 7-12 weeks. Others define it as pain that lasts beyond the expected period of healing, and acknowledge that chronic pain may not have well-defined underlying pathological causes. Others classify frequently recurring back pain as chronic pain since it intermittently affects an individual over a long period. Most national insurance and industrial sources of data include only those individuals in whom symptoms result in loss of days at work or other disability. Thus, even less is known about the epidemiology of chronic low-back pain with no associated work disability or compensation. Chronic low-back pain has also become a diagnosis of convenience for many people who are actually disabled for socioeconomic, work related, or psychological reasons. In fact, some people argue that chronic disability in back pain is primarily related to a psychosocial dysfunction. Because the validity and reliability of some of the existing data are uncertain, caution is needed in an assessment of the information on this type of pain. PMID- 10470717 TI - The burden of disease among the global poor. AB - BACKGROUND: Global and regional estimates show that non-communicable diseases in old age are rising in importance relative to other causes of ill health as populations age, and as progress continues against communicable diseases among infants and children. However, these estimates, which cover population groups at all income levels, do not accurately reflect conditions that prevail among the poor. We estimated the burden of disease among the 20% of the global population living in countries with the lowest per capita incomes, compared with the 20% of the world's people living in the richest countries. METHODS: Estimates for the global poorest and richest 20% were prepared for 1990 for deaths and disability adjusted life years (DALYs), by a procedure used in a prominent recent study of the global disease burden. Projected mortality rates in the year 2020 were established for the world's poorest and richest 20% under various assumptions about the future rate of decline in communicable and non-communicable diseases. FINDINGS: In 1990, communicable diseases caused 59% of death and disability among the world's poorest 20%. Among the world's richest 20%, on the other hand, non communicable diseases caused 85% of death and disability. A raised baseline rate of communicable disease decline between 1990 and 2020 would increase life expectancy among the world's poorest 20% around ten times as much as it would the richest 20% (4.1 vs 0.4 years). However, the poorest 20% would gain only around a quarter to a third as much as the richest 20% from a similar increase in non communicable diseases (1.4 vs 5.3 years). As a result, a faster decline in communicable diseases would decease the poor-rich gap in 2020, but under an accelerated rate of overall decline in non-communicable diseases, the poor-rich gap would widen. INTERPRETATION: Our estimates are crude, but despite their limitations, they give a more accurate picture of changes in attributable mortality among the world's poor than do the global averages in current use. PMID- 10470718 TI - Magnitude of the problem of childhood pneumonia. PMID- 10470719 TI - What is hypertension? PMID- 10470720 TI - What is hypertension? PMID- 10470721 TI - Laparoscopically assisted abdominal aortic aneurysm repair. PMID- 10470722 TI - Laparoscopically assisted abdominal aortic aneurysm repair. PMID- 10470723 TI - Episiotomy: a form of genital mutilation. PMID- 10470724 TI - Estimation of mean aortic pressure. PMID- 10470725 TI - Iodine deficiency in Italy. Italian Study Group on Urinary Iodine of SIEDP. PMID- 10470726 TI - Corneodesmosin (MHC S) gene in guttate psoriasis. PMID- 10470727 TI - From effect size into number needed to treat. PMID- 10470728 TI - From effect size into number needed to treat. PMID- 10470729 TI - Transit-time analysis in hepatic cirrhosis. PMID- 10470730 TI - Endotoxins in chronic heart failure. PMID- 10470731 TI - Sleep apnoea. PMID- 10470732 TI - UKPDS: what was the question? UK Prospective Diabetes Study. PMID- 10470733 TI - Medical achievements under civil war conditions. PMID- 10470734 TI - Scientific dishonesty. PMID- 10470735 TI - Slow drip of progress. PMID- 10470736 TI - Sociological models and health care. PMID- 10470737 TI - Funding of clinical research. PMID- 10470738 TI - Mikhail Bulgakov's myth about medicine, literature, and fiction. PMID- 10470739 TI - Centenarian scientists. PMID- 10470740 TI - A tangled history of America's relationship with illegal drugs. PMID- 10470741 TI - The Nobel Chronicles. 1967: George Wald (1906-97); Ragnar A Granit (1900-91); and Haldan Keffer Hartline (1903-83). PMID- 10470742 TI - "I'm off tomorrow. What do I need to take?". PMID- 10470743 TI - Effect of enteral feeding with eicosapentaenoic acid, gamma-linolenic acid, and antioxidants in patients with acute respiratory distress syndrome. Enteral Nutrition in ARDS Study Group. AB - OBJECTIVES: Recent studies in animal models of sepsis-induced acute respiratory distress syndrome (ARDS) have shown that a low-carbohydrate, high-fat diet combining the anti-inflammatory and vasodilatory properties of eicosapentaenoic acid (EPA; fish oil), gamma-linolenic acid (GLA; borage oil) (EPA+GLA), and antioxidants improves lung microvascular permeability, oxygenation, and cardiopulmonary function and reduces proinflammatory eicosanoid synthesis and lung inflammation. These findings suggest that enteral nutrition with EPA+GLA and antioxidants may reduce pulmonary inflammation and may improve oxygenation and clinical outcomes in patients with ARDS. DESIGN: Prospective, multicentered, double-blind, randomized controlled trial. SETTING: Intensive care units of five academic and teaching hospitals in the United States. PATIENTS: We enrolled 146 patients with ARDS (as defined by the American-European Consensus Conference) caused by sepsis/pneumonia, trauma, or aspiration injury in the study. INTERVENTIONS: Patients meeting entry criteria were randomized and continuously tube-fed either EPA+GLA or an isonitrogenous, isocaloric standard diet at a minimum caloric delivery of 75% of basal energy expenditure x 1.3 for at least 4 7 days. MEASUREMENTS AND MAIN RESULTS: Arterial blood gases were measured, and ventilator settings were recorded at baseline and study days 4 and 7 to enable calculation of PaO2/FIO2, a measure of gas exchange. Pulmonary neutrophil recruitment was assessed by measuring the number of neutrophils and the total cell count in bronchoalveolar lavage fluid at the same time points. Clinical outcomes were recorded. Baseline characteristics of 98 evaluable patients revealed that key demographic, physiologic, and ventilatory variables were similar at entry between both groups. Multiple bronchoalveolar lavages revealed significant decreases (approximately 2.5-fold) in the number of total cells and neutrophils per mL of recovered lavage fluid during the study with EPA+GLA compared with patients fed the control diet. Significant improvements in oxygenation (PaO2/FIO2) from baseline to study days 4 and 7 with lower ventilation variables (FIO2, positive end-expiratory pressure, and minute ventilation) occurred in patients fed EPA+GLA compared with controls. Patients fed EPA+GLA required significantly fewer days of ventilatory support (11 vs. 16.3 days; p = .011), and had a decreased length of stay in the intensive care unit (12.8 vs. 17.5 days; p = .016) compared with controls. Only four of 51 (8%) patients fed EPA+GLA vs. 13 of 47 (28%) control patients developed a new organ failure during the study (p = .015). CONCLUSIONS: The beneficial effects of the EPA+GLA diet on pulmonary neutrophil recruitment, gas exchange, requirement for mechanical ventilation, length of intensive care unit stay, and the reduction of new organ failures suggest that this enteral nutrition formula would be a useful adjuvant therapy in the clinical management of patients with or at risk of developing ARDS. PMID- 10470744 TI - A 25-year study of nosocomial bacteremia in an adult intensive care unit. AB - OBJECTIVE: To identify the organisms, their antibiotic susceptibility, and the associated focus on infection causing nosocomial bacteremia in patients in an adult intensive care unit (ICU) between 1971 and 1995. DESIGN: Prospective observational study. SETTING: A 12-bed general adult ICU in a 1,000-bed tertiary referral teaching hospital. PATIENTS: Four hundred eighty-six episodes of bacteremia involving 570 organisms in 425 patients. MEASUREMENTS AND MAIN RESULTS: Blood cultures taken from patients with suspected nosocomial infection were analyzed. Isolated organisms were identified, and their susceptibility to commonly used antibiotics was determined. Clinical details, including antibiotic treatment, were recorded for all patients. From 1986 to 1995, culture results of samples obtained from other sites were used to help identify the focus of infection causing bacteremia. All results were collected prospectively by clinical microbiologists. Between 1971 and 1990, the number of bacteremias and the relative frequency of isolation of individual organisms changed little, with Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella species predominating. During 1991 to 1995, the number of bacteremias increased two-fold, largely attributable to increased isolation of Enterococcus species, coagulase-negative staphylococci, intrinsically antibiotic-resistant gram negative organisms (particularly P. aeruginosa), and Candida species. The most commonly used antibiotics for the treatment of bacteremic patients throughout the 1970s were amoxicillin and gentamicin. After the introduction of cephalosporins in the early 1980s, their use increased progressively to equal that of gentamicin in the 1990s, whereas amoxicillin use decreased. Since the introduction of cephalosporins, increases in the antibiotic resistance of gram-negative organisms have been largely confined to an outbreak of gentamicin- and ceftazidime resistant organisms caused by contaminated arterial pressure monitors during 1992 and 1993 and a two-fold increase in ceftazidime resistance of the Pseudomonas species. Gentamicin resistance of gram-negative aerobes remained unchanged (excluding the arterial pressure monitor outbreak), despite gentamicin being one of the most frequently prescribed antibiotics throughout the 25-yr period. Between 1986 and 1995, two thirds of all bacteremic organisms were cultured from intravascular catheters, which were designated as the focus of infection, 7% were secondary to gastrointestinal pathology, but only approximately 3% were secondary to wound, respiratory tract, or urinary tract infections. CONCLUSIONS: Bacteremias have become more frequent in the ICU, probably because of the increased use of intravascular catheters, which are the most frequent foci for bacteremic infection. The spectrum of organisms has changed, and this can be temporally related to the changes in the antibiotics prescribed. Gentamicin resistance of gram-negative organisms has not increased during a 25-yr period, despite being one of the most frequently prescribed antibiotics in the ICU. PMID- 10470745 TI - Acidic conditions exacerbate interferon-gamma-induced intestinal epithelial hyperpermeability: role of peroxynitrous acid. AB - OBJECTIVE: Nitric oxide (NO*) derived from exogenous donors has been shown to increase the permeability of cultured intestinal epithelial monolayers, an effect that is augmented by mildly acidic conditions. Because interferon-gamma (IFN gamma) also increases intestinal epithelial permeability, at least partly by an NO*-dependent mechanism, we sought to determine whether IFN-gamma-induced hyperpermeability is increased under acidic conditions. METHODS: Human intestinal epithelial (Caco-2BBe) cells were grown as monolayers on permeable supports in bicameral chambers. Permeability was assessed by measuring transepithelial electrical resistance (TER) or the transepithelial passage of fluorescein disulfonic acid. Inducible nitric oxide synthase (iNOS) messenger RNA expression was determined by northern blot analysis. Concentrations of nitrite and nitrate (NO2-/NO3-), stable oxidation products of NO*, were determined using the Greiss reaction. Cellular adenosine triphosphate (ATP) levels were determined using the luciferin/luciferase method. MEASUREMENTS AND MAIN RESULTS: Incubation of Caco 2BBe monolayers with INF-gamma (1000 units/mL) at an extracellular pH (pHo) of 7.4 increased permeability to fluorescein disulfonic acid and decreased TER. However, incubation of monolayers with IFN-gamma under mildly acidic conditions (i.e., pHo 7.0-6.6) accelerated the decrease in TER and augmented the increase in permeability induced by the cytokine. IFN-gamma-induced iNOS messenger RNA expression and NO2-/NO3- accumulation in medium were unaffected by acidic conditions. At pHo 7.4, incubation of Caco-2BBe monolayers with IFN-gamma (1000 units/mL) for 72 hrs had no effect on intracellular ATP content compared with monolayers simultaneously incubated under the same conditions but in the absence of the cytokine. However, when the cells were incubated for 72 hrs with the same concentration of IFN-gamma under mildly acidic conditions (i.e., pHo 7.0 or 6.6), ATP levels were significantly decreased. At pHo 7.0, IFN-gamma-induced increases in permeability were ameliorated by addition of the following agents: 2-phenyl 4,4,5,5- tetramethylimidazoline-1-oxyl-3-oxide (a NO* scavenger), N(G)-monomethyl L-arginine (a iNOS inhibitor), dimethyl sulfoxide (a hydroxyl radical scavenger), and ascorbate (a peroxynitrous acid scavenger). CONCLUSION: Mild acidosis augments IFN-gamma-induced intestinal epithelial hyperpermeability and ATP depletion, possibly by fostering the formation of peroxynitrous acid and/or hydroxyl radical. PMID- 10470746 TI - Kerosene-induced severe acute respiratory failure in near drowning: reports on four cases and review of the literature. AB - OBJECTIVE: The purpose of this study is to present an unusual respiratory and cardiovascular course after intoxication and near drowning in a river contaminated with kerosene. DESIGN: Case reports and review of the literature. SETTING: Intensive care unit of a university-affiliated hospital. PATIENTS: Four patients after near drowning. INTERVENTION: Supportive only. RESULTS: The four patients developed acute respiratory failure. Cardiomyopathy was present in three patients and a persistent hypokalemia in two patients. The onset of the symptoms was delayed, which led to underestimation of the severity of their illness. Two of the four patients died. The diagnosis of hydrocarbon intoxication was based on bronchoalveolar lavage results, neutrophilic alveolitis with the presence of lipid-laden macrophages, and evidence of lipoid pneumonia from the autopsy performed on one victim. One patient who clinically deteriorated and another who developed a severe restrictive pulmonary disorder were treated with corticosteroids, which were effective only in the latter patient. CONCLUSIONS: Acute kerosene intoxication in a near-drowning event often results in severe respiratory and cardiac failure, with a high fatality rate. Treatment with corticosteroids may lead to a rapid improvement in lung function. PMID- 10470747 TI - Local effect of lung contusion on lung surfactant composition in multiple trauma patients. AB - OBJECTIVE: The aim of this study was to investigate the direct influence of lung contusion on pulmonary surfactant in multiple trauma patients. DESIGN: Prospective, nonrandomized study. SETTING: University hospital, trauma intensive care unit. PATIENTS: Eighteen multiple trauma patients with unilateral lung contusions and Injury Severity Scores >19 were studied prospectively. INTERVENTIONS: Bronchoalveolar lavage was performed daily until either day 7 or extubation. Samples from the side of lung contusion (n = 62) and the contralateral, uninjured side (n = 62) were obtained at the same time in 14 patients. Total phospholipids, total phospholipid classes, and surfactant apoprotein A were quantified. Additionally, surfactant function was measured with a pulsating bubble surfactometer in four patients. All data are presented as mean +/- SEM. Statistical analyses were performed using programs of SPSS for Windows 6.1.3 (SPSS Inc., Chicago, IL) (Student's t-test; p < .05). MEASUREMENTS AND MAIN RESULTS: Total phospholipids were significantly increased on the side of lung contusion (contusion side, 40+/-7 microg/mL; contralateral side, 21+/-3 microg/mL; p = .004). The percentage contents of phosphatidylcholine (contusion side, 87.1%+/-1.0%; contralateral side, 84.3%+/-1.0%; p = .04) and sphingomyelin (contusion side, 2.9%+/-0.3%; contralateral side, 1.9%+/-0.2%; p = .004) were significantly higher. In contrast, the percentage content of phosphatidylglycerol was significantly decreased (contusion side, 4.1%+/-0.1%; contralateral side, 6.9%+/-0.6%; p = .001). No alterations were found for the relative contents of phosphatidylethanolamine (contusion side, 2.4%+/-0.2%; contralateral side, 2.2%+/ 0.2%; p = .47), phosphatidylinositol (contusion side, 3.5%+/-0.4%; contralateral side, 4.6%+/-0.5%; p = .06), and surfactant apoprotein A (contusion side, 7177+/ 1404 ng/mL; contralateral side, 4513+/-787 ng/mL, p = .10). There was no statistical difference for minimal surface tension measured with the pulsating bubble surfactometer after 5 mins of oscillation (contusion side, 29.5+/-2.3 mN/m; contralateral side, 23.7+/-2.1 mN/m; p = .08). CONCLUSIONS: Direct damage of lung parenchyma by lung contusion alters the composition of surfactant. No additional changes in surfactant function were observed that would argue in favor of functional compensation. PMID- 10470748 TI - Enteral nutrition-related gastrointestinal complications in critically ill patients: a multicenter study. The Nutritional and Metabolic Working Group of the Spanish Society of Intensive Care Medicine and Coronary Units. AB - OBJECTIVE: To evaluate the frequency of gastrointestinal complications (GICs) in a prospective cohort of critically ill patients receiving enteral nutrition and to assess its effect on nutrient administration and its relationship to the patient's outcome. DESIGN: Multicenter, prospective cohort study. SETTING: Thirty seven multidisciplinary intensive care units (ICUs) in Spain. PATIENTS: Prospective cohort of 400 consecutive patients admitted to the ICU and receiving enteral nutrition. INTERVENTIONS: Noninterventional, follow-up study. MEASUREMENTS AND MAIN RESULTS: Enteral nutrition-related GICs and their management were defined by consensus before data collection. A set of variables related to enteral nutrition administration and the presence of GICs was recorded. During the 1-month study period, 400 patients were enrolled, and a total of 3,778 enteral feeding days were analyzed. The mean time of enteral nutrition was 9.6+/-0.4 days. Mean elapsed time from ICU admission to the start of enteral feeding was 3.1+/-0.2 days. A total of 265 patients (66.2%) received a standard polymeric formula, and 132 (33.8%) received a disease-specific one. Enteral feeds were administered mainly through a nasogastric tube (91%). One or more GICs were presented by 251 patients (62.8%) during the feeding course. The frequency of each particular GIC was as follows: high gastric residuals, 39%; constipation, 15.7%; diarrhea, 14.7%; abdominal distention, 13.2%; vomiting, 12.2%; and regurgitation, 5.5%. Enteral nutrition withdrawal as a consequence of noncontrollable GICs occurred in 15.2% of patients. The volume ratio (expressed as the ratio between administered and prescribed volumes) was calculated daily and was used as an index of diet administration efficacy. Patients with GICs had a lower volume ratio than did patients without GICs (63.1+/-1.20% vs. 93.3+/ 0.3%) (p < .001), a longer length of stay (20.6+/-1.2 vs. 15.2+/-1.3 days) (p < .01), and higher mortality (31% vs. 16.1%) (p < .001). CONCLUSIONS: The frequency of enteral nutrition-related GICs in critically ill patients is high. High gastric residuals is the most frequent GIC. These complications decreased nutrient intake and, if persistent, could expose the patients to undernutrition. Enteral feeding gastrointestinal intolerance seems to have an evolutive effect in prolonging the ICU stay and increasing patient mortality. PMID- 10470749 TI - Time-course of impairment of respiratory mechanics after cardiac surgery and cardiopulmonary bypass. AB - OBJECTIVE: Cardiopulmonary bypass (CPB) is associated with abnormalities of lung function characterized by an increase in static elastance of the respiratory system. We examined the following: a) the effects of CPB on the total inspiratory volume-pressure (V-P) relationship of the respiratory system; b) the relative contribution of the chest wall and lung to the impairment of respiratory system mechanics; and c) the time-course of such impairment. DESIGN: Prospective, interventional study. SETTING: Surgical and medical intensive care units in a teaching hospital. PATIENTS: Eight adult patients scheduled for elective open heart surgery to correct valvular dysfunction. INTERVENTIONS: V-P curves (interrupter technique) of the respiratory system were partitioned between the chest wall and lung by measurements of esophageal pressure. Measurements were obtained before sternotomy (control), immediately after, 4 hrs after, and 7 hrs after separation from CPB. MEASUREMENTS AND MAIN RESULTS: Control V-P relationships of the respiratory system and lung showed lower inflection points (Pflex) at pressure values of 5.9+/-2.3 and 4.3+/-2.5 cm H2O, respectively. Immediately after and 4 hrs after separation from CPB, both curves had sigmoid shapes because of lower Pflex and formation of upper inflection (UIP) points. The pressures corresponding to the Pflex increased significantly (p < .001) by 56%+/ 3% and 78%+/-4%, whereas the UIP corresponded to a pressure value of 42.34+/-8.5 and 35.6+/-7.8 cm H2O in the respiratory system and lung, respectively. A linear V-P relationship of the chest wall was observed during the control condition and after separation from CPB. Four hours later, no further increases in the pressure values corresponding to Pflex were observed on the inspiratory V-P curves of the respiratory system and lung, whereas the UIP occurred at a pressure of 35.6+/-9.1 and 29.7+/-8.4 cm H2O, respectively. A UIP was present on the V-P curve of the chest wall at a volume of 0.77+/-0.02 L. Seven hours after separation from CPB, the inspiratory V-P curves of the respiratory system, chest wall, and lung returned to normal. CONCLUSIONS: Sternotomy and CPB produced immediate changes in lung mechanics. Chest wall mechanics were affected only 4 hrs after sternotomy. Seven hours after disconnection from CPB, all mechanics had returned to normal. PMID- 10470751 TI - Evaluation of patient-perceived health status using the Medical Outcomes Survey Short-Form 36 in an intensive care unit population. AB - OBJECTIVE: Baseline patient functional status as assessed by providers is correlated with mortality after intensive care unit (ICU) admission. We wanted to see if patient self-perception of health status before admission to an ICU correlated with functional outcome. DESIGN: Prospective survey on a convenience sample. SETTING: Single urban university-affiliated Veterans Affairs Medial Center. PATIENTS: One hundred ninety-nine patients in surgical and medical/coronary ICUs. INTERVENTIONS: None. MEASUREMENTS: Patient-assessed baseline health status was monitored with the Medical Outcome Survey Short-Form 36 (SF-36), consisting of 36 questions that evaluate eight health status concepts. In addition, baseline functional status (Zubrod scale) was determined and severity of illness (Acute Physiology and Chronic Health Evaluation [APACHE] II) data were collected. Zubrod functional status, which includes mortality, was determined 6 wks and 6 months after ICU admission, and correlation coefficients were calculated. MAIN RESULTS: We found it feasible to collect SF-36 health status data on a 9% sample in this setting. Less than 1% of responses were completed by proxy. The SF-36 data were internally consistent, and several of its scales including general health perception and physical functioning correlated with patient Zubrod functional status (r2 = .08, p < .001; r2 = .14, p < .001) at 6 wks as did vitality (r2 = .04, p < .01), social function (r2 = .03, p < .05), and physical role function (r2 = .02, p = .053), although to a lesser extent. Similar correlations were also found with 6-month functional status. CONCLUSIONS: We conclude that use of the SF-36 is time efficient in an ICU setting and correlates with 6-wk and 6-month functional outcome. It correlates as well with functional outcome as either the baseline Zubrod functional status or the APACHE II severity of illness measurement. The five-question general health evaluation portion correlated almost as well with outcome as the more extensive 36-item questionnaire. Use of the SF-36 may define patient populations for comparison across hospitals. It may also target individuals with needs for additional posthospitalization care, including rehabilitation services or nursing home placement. PMID- 10470750 TI - Continuous infusion of lorazepam versus medazolam in patients in the intensive care unit: sedation with lorazepam is easier to manage and is more cost effective. AB - OBJECTIVE: To evaluate the effectiveness of lorazepam and midazolam for long-term sedation of critically ill, mechanically ventilated patients. DESIGN: Double blind, randomized, controlled study. SETTING: Medical intensive care unit in a university teaching hospital. PATIENTS: Sixty-four evaluable adult patients admitted to the intensive care department requiring mechanical ventilation for >3 days. INTERVENTIONS: Patients were randomized to receive blinded solutions of either lorazepam or midazolam by continuous infusion. The lowest dose that achieved an adequate sedation was infused. The maximum dose allowed for each drug was 60 mg/hr for midazolam and 4 mg/hr for lorazepam. Sedation was assessed initially and at least every 8 hrs thereafter on a seven-point scale if the sedation was adequate and every 2 hrs if it was not. MEASUREMENTS AND MAIN RESULTS: Measurements included the score on the sedation scale, the time between determination of the desired level of sedation and the achievement of that level, and plasma concentrations. It is significantly easier to reach a desired level of sedation with lorazepam than with midazolam. No difference in recovery was found in the 24 hrs after discontinuation of therapy. The fact that there are many factors influencing midazolam pharmacokinetics may explain the more difficult management of desired sedation levels. The equipotent dose of 10 mg of midazolam proved to be 0.7 mg of lorazepam in long-term sedation. The average cost for therapy with midazolam was approximately ten times more than that with lorazepam. CONCLUSIONS: Lorazepam is a useful alternative to midazolam for the long-term sedation of patients in the medical intensive care unit and provides easier management of the sedation level. Sedation with lorazepam offers a significant cost-savings. PMID- 10470752 TI - Use of single-frequency bioimpedance at 50 kHz to estimate total body water in patients with multiple organ failure and fluid overload. AB - OBJECTIVE: To investigate the relationship between single-frequency bioimpedance at 50 kHz (both total body impedance and segmental impedance) and total body water, measured using tritiated water in the presence of the severe fluid retention seen in multiple organ failure. DESIGN: Prospective, controlled study. SETTING: General intensive care unit. SUBJECTS: Twenty patients with multiple organ failure and 30 normal volunteers, of whom a subgroup of ten had total body water measured. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Total body water and total and segmental bioimpedance values at 50 kHz were measured using tritiated water and a Holtain body composition analyzer in 20 patients with multiple organ failure and in ten normal volunteers. An additional 20 normal volunteers also had segmental and total body impedance measured. There was no difference in the linear regression lines constructed for the patients and the volunteers, but the SEM in the patients (7.6 L) was four times that seen in the normal subjects (1.9 L). In a further seven patients, the impedance technique overestimated the change in total body water, deduced from acute changes in weight, by between 0% and 46% (median, 12%). In the patients, who were supine, the knee-ankle segment contributed significantly more to total body impedance than it did in the normal volunteers (42.5% vs. 34.4%; p < .001), who were upright and mobile immediately before the measurement. CONCLUSIONS: Although single-frequency bioimpedance does give an indication of total body water and change in total body water, it is neither precise nor accurate enough to be the sole guide to fluid therapy. The proportion of total impedance contributed by the knee-ankle segment, which contains relatively little water, was significantly greater in the patients than in the controls, probably reflecting better drainage of fluid from the lower limb in the supine position. PMID- 10470753 TI - Liposomal prostaglandin E1 (TLC C-53) in acute respiratory distress syndrome: a controlled, randomized, double-blind, multicenter clinical trial. TLC C-53 ARDS Study Group. AB - OBJECTIVE: To evaluate the safety and efficacy of an intravenous liposomal dispersion of prostaglandin E1 as TLC C-53 in the treatment of patients with acute respiratory distress syndrome (ARDS). DESIGN: Randomized, prospective, multicenter, double-blind, placebo-controlled, phase III clinical trial. SETTING: Forty-seven community and university-affiliated hospitals in the United States. PATIENTS: A total of 350 patients with ARDS were enrolled in this clinical trial. INTERVENTION: Patients were prospectively randomized in a 1:1 ratio to receive either liposomal prostaglandin E1 or placebo. The study drug was infused intravenously for 60 mins every 6 hrs for 7 days starting with a dosage of 0.15 microg/kg/hr. The dose was increased every 12 hrs until the maximal dose (3.6 microg/kg/hr) was attained or intolerance to further increases developed. Patients received standard aggressive medical/surgical care during the infusion period. OUTCOME MEASURES: The primary outcome measure was the time it took to wean the patient from the ventilator. Secondary end points included time to improvement of the PaO2/FIO2 ratio (defined as first PaO2/FIO2 > 300 mm Hg), day 28 mortality, ventilator dependence at day 8, changes in PaO2/FIO2, incidence of and time to development/resolution of organ failure other than ARDS. RESULTS: A total of 348 patients could be evaluated for efficacy. The distribution of variables at baseline describing gender, lung injury scores, Acute Physiology and Chronic Health Evaluation II scores, PaO2/FIO2, pulmonary compliance, and time from onset of ARDS or from institution of mechanical ventilation to the first dose of study drug was similar among patients in the liposomal prostaglandin E1 (n = 177) and the placebo (n = 171) treatment arms. There was no significant difference in the number of days to the discontinuation of ventilation in the liposomal prostaglandin E1 group compared with the placebo group (median number of days to off mechanical ventilation, 16.9 in patients receiving liposomal prostaglandin E1 and 19.6 in those administered placebo; p = .94). Similarly, mortality at day 28 was not significantly different in the two groups (day 28 mortality, 57 of 176 (32%) in the liposomal prostaglandin E1 group and 50 of 170 (29%) in patients receiving placebo; p = .55). In contrast, treatment with liposomal prostaglandin E1 was associated with a significantly shorter time to reach a PaO2/FIO2 ratio of >300 mm Hg (median number of days to reaching a PaO2/FIO2 ratio >300 mm Hg: 9.8 days in the liposomal prostaglandin E1 group and 13.7 days in patients receiving the placebo; p = .02). Among the subgroups examined, time to off mechanical ventilation was significantly reduced in patients who received at least 85% of a full dose (i.e., > 45.9 microg/kg) of liposomal prostaglandin E1 (median number of days to discontinuation of ventilation, 10.3 in the liposomal prostaglandin E1 group and 16.3 days in patients receiving placebo; p = .05). The overall incidence of serious adverse events was not significantly different in the liposomal prostaglandin E1 (40%) or placebo-treated (37%) groups. Drug-related adverse events of all kinds were reported in 69% of the patients receiving liposomal prostaglandin E1 compared with 33% of the placebo group, with hypotension and hypoxia (occurring in 52% and 24% of the liposomal prostaglandin E1-treated patients, respectively, and 17% and 5% of the placebo-treated patients, respectively) being noted most frequently. CONCLUSIONS: In the intent-to-treat population of patients with ARDS, treatment with liposomal prostaglandin E1 accelerated improvement in indexes of oxygenation but did not decrease the duration of mechanical ventilation and did not improve day 28 survival. PMID- 10470754 TI - Evaluation of an interdisciplinary data set for national intensive care unit assessment. AB - OBJECTIVES: To evaluate the ability of an interdisciplinary data set (recently defined by the Austrian Working Group for the Standardization of a Documentation System for Intensive Care [ASDI]) to assess intensive care units (ICUs) by means of the Simplified Acute Physiology Score II (SAPS II) for the severity of illness and the simplified Therapeutic Intervention Scoring System (TISS-28) for the level of provided care. DESIGN: A prospective, multicentric study. SETTING: Nine adult medical, surgical, and mixed ICUs in Austria. PATIENTS: A total of 1234 patients consecutively admitted to the ICUs. INTERVENTIONS: Collection of data for the ASDI data set. MEASUREMENTS AND MAIN RESULTS: The overall mean SAPS II score was 33.1+/-2.1 points. SAPS II overestimated hospital mortality by predicting mortality of 22.2%+/-2.9%, whereas observed mortality was only 16.8%+/ 2.2%. The Hosmer-Lemeshow goodness-of-fit test for SAPS II scores showed lacking uniformity of fit (H = 53.78, 8 degrees of freedom; p < .0001). TISS-28 scores were recorded on 8616 days (30.6+/-1.5 points). TISS-28 scores were higher in nonsurvivors than in survivors (30.4+/-0.9 vs. 25.7+/-0.4, respectively; p < .05). No significant correlation between mean TISS-28 per patient per unit on the day of admission and mean predicted hospital mortality (r2 = .23; p < .54) or standardized mortality ratio per unit (r2 = -.22; p < .56) was found. CONCLUSIONS: Implementation of an interdisciplinary data set for ICUs provided data with which to evaluate performance in terms of severity of illness and provided care. The SAPS II did not accurately predict outcomes in Austrian ICUs and must, therefore, be customized for this population. A combination of indicators for both severity of illness and amount of provided care is necessary to evaluate ICU performance. Further data acquisition is needed to customize the SAPS II and to validate the TISS-28. PMID- 10470755 TI - Prospective, randomized, controlled clinical trial comparing traditional versus reduced tidal volume ventilation in acute respiratory distress syndrome patients. AB - OBJECTIVE: To assess the safety and potential efficacy of a mechanical ventilation strategy designed to reduce stretch-induced lung injury in acute respiratory distress syndrome. DESIGN: Prospective, randomized, controlled clinical trial. SETTING: Eight intensive care units in four teaching hospitals. PATIENTS: Fifty-two patients with acute respiratory distress syndrome. INTERVENTIONS: Traditional tidal volume patients: tidal volume 10-12 mL/kg ideal body weight, reduced if inspiratory plateau pressure was > 55 cm H2O (7.3 kPa). Small tidal volume patients: tidal volume 5-8 mL/kg ideal body weight, to keep plateau pressure < 30 cm H2O (4.0 kPa). MEASUREMENTS AND MAIN RESULTS: Mean tidal volumes during the first 5 days in traditional and small tidal volume patients were 10.2 and 7.3 mL/kg, respectively (p < .001), with mean plateau pressure = 30.6 and 24.9 cm H2O (3.3 kPa), respectively (p < .001). There were no significant differences in requirements for positive end-expiratory pressure or FIO2, fluid intakes/outputs, requirements for vasopressors, sedatives, or neuromuscular blocking agents, percentage of patients that achieved unassisted breathing, ventilator days, or mortality. CONCLUSIONS: The reduced tidal volume strategy used in this study was safe. Failure to observe beneficial effects of small tidal volume ventilation treatment in important clinical outcome variables may have occurred because a) the sample size was too small to discern small treatment effects; b) the differences in tidal volumes and plateau pressures were modest; or c) reduced tidal volume ventilation is not beneficial. PMID- 10470756 TI - Assessing sedation during intensive care unit mechanical ventilation with the Bispectral Index and the Sedation-Agitation Scale. AB - OBJECTIVE: To describe the level of sedation for a cohort of mechanically ventilated adult intensive care unit (ICU) patients using validated subjective and objective tools. DESIGN: Prospective convenience sample. SETTING: Multidisciplinary 34-bed ICU at Maine Medical Center, a 599-bed nonuniversity, academic medical center. PATIENTS: Sixty-three adult ICU patients were monitored during 64 episodes of ventilatory support. MEASUREMENTS AND MAIN RESULTS: Patients were prospectively evaluated by one trained investigator using the revised Sedation-Agitation Scale (SAS) and were simultaneously monitored for 1 to 5 hrs using the Bispectral Index (BIS), a numeric scale from 0 to 100 derived from the electroencephalogram. BIS values were assigned to baseline, stimulated, and average conditions for each patient by a separate investigator blinded to SAS scores. Ventilator settings, medications, and the lung injury severity (LIS) score were also recorded. Sedation levels varied from very deep sedation (SAS score = 1, BIS score = 43) to mild agitation (SAS score = 5, BIS score = 100). Heavily sedated patients (SAS score = 1-2, n = 20) had higher FIO2 (0.52 vs. 0.42, p = .008), oxygenation index (9.4 vs. 5.4, p = .03), and LIS scores (1.3 vs. 0.7, p = .004) and lower baseline (66 vs. 78, p = .01), average (66 vs. 81, p < .001), and stimulated (89 vs. 96, p = .016) BIS scores compared with more awake patients. Patients with intermittent neuromuscular blockade use (n = 4) had higher FIO2 (0.65 vs. 0.44, p = .006), minute ventilation (14.6 vs. 9.9 L/min, p = .005), positive end-expiratory pressure (7.5 vs. 4.8 cm H2O, p = .05), oxygenation index (15.7 vs. 6.0, p < .001), and LIS scores (3.3 vs. 1.0, p = .036) and were more sedated, with higher suppression ratios (3.5 vs. 0.6, p = .05) and lower SAS scores (1.5 vs. 4, p = .035). The average BIS values correlated well with SAS (r2 = .21, p < .001). CONCLUSIONS: SAS and BIS work well to describe the depth of sedation for ventilated ICU patients. Deeper sedation and intermittent neuromuscular blockade were used for patients with greater ventilatory requirements and more severe lung disease. The correlation between subjective and objective scales varied in medical, surgical, and trauma patients. Further research with SAS and BIS may facilitate the development of quantitative sedation guidelines for the ICU. PMID- 10470757 TI - Semicontinuous cardiac output monitoring using a neural network. AB - OBJECTIVES: This study compared 2-mL bolus thermodilution cardiac output measurements with standard 10-mL bolus measurements. DESIGN: Cardiac output was measured with the new 2-mL bolus technique and the 10-mL standard thermodilution technique in a perspective series. We describe a system that automatically cools and injects 2-mL boluses of saline into a standard pulmonary artery catheter. It uses a Peltier effect solid-state cooler and pneumatically driven syringe injector to measure cardiac output once per minute. SETTING: Animal laboratory. ANIMALS: Eight adult Duroc swine weighing between 38.0 and 57.5 kg. INTERVENTIONS: Once each minute, 2 mL of cooled 5% dextrose was injected through the pulmonary catheter. Once every 8 mins, four sequential measurements of cardiac output were made using 10-mL injections. MEASUREMENTS AND MAIN RESULTS: A total of 1249 paired waveforms were processed with both a conventional algorithm and with a neural network. For the conventional algorithm, the correlation coefficient was r2 = .92 and the SD of the difference was 1.30 L/min. For the neural network, the correlation coefficient was r2 = .94 and the SD of the difference was 0.88 L/min. Output filtering improved the results in both cases. CONCLUSION: Neural networks accurately derive cardiac output from 2-mL bolus thermodilution injections, allowing cardiac output to be monitored automatically once per minute in many patients. The technique is convenient and uses standard low-cost catheters. PMID- 10470758 TI - Bacterial dissemination, rather than translocation, mediates hypermetabolic response in endotoxemic rats. AB - OBJECTIVE: To study the pathogenesis of the host response during bacterial translocation, a rat model was designed for prolonged follow-up after injury. DESIGN: A prospective, controlled animal study. SETTING: Animal laboratory. SUBJECTS: Young male Wistar rats. INTERVENTIONS: Antibiotic decontamination of rats was performed 4 days before intragastric inoculation with a selected Escherichia coli strain (10(10) bacteria/kg of body weight). Two days later, the rats received a lipopolysaccharide injection or not (control group) and were observed for 3 days. They were then killed. A reference group (pair-fed healthy animals) was studied in parallel. MEASUREMENTS AND MAIN RESULTS: During observations, urinary total nitrogen loss and 3-methylhistidine excretion were determined daily. When the rats were killed, mesenteric lymph nodes (MLNs), spleen, and liver were aseptically removed and cultured. Colonies identified as translocated E. coli were counted in each organ. Intracellular amino acid free pools were measured in extensor digitorum longus and anterior tibialis. Endotoxin induces bacterial translocation of bacteria from gut lumen to MLNs (100% vs. 59% in the lipopolysaccharide-untreated control group; p < .05) and dissemination to spleen and liver (65% and 45% of positive cultures after endotoxemia, respectively, vs. 6% and 12% in the control groups). No translocation occurred in the reference group. Evidence for the hypermetabolic response was seen in lipopolysaccharide-treated and infected rats, but protein catabolism was more closely related to the occurrence of bacterial dissemination to spleen and liver than to translocation alone (e.g., the cumulative 3-methylhistidine excretion during the observation period was 4.07+/-0.18 micromol in uninfected rats, 4.48+/ 0.29 in rats with positive MLN cultures alone and 6.17+/-0.30 in MLN, spleen, or liver infected rats; 1 vs. 2, NS; 3 vs. 1, and 3 vs. 2, p < .05). CONCLUSIONS: Gut barrier failure is associated with a deep excessive catabolic response in the host. The mechanism by which the metabolic state affects the resistance to infection apparently involves amino acid metabolism. PMID- 10470759 TI - Protective effect of poly(ADP-ribose) synthetase inhibition on multiple organ failure after zymosan-induced peritonitis in the rat. AB - BACKGROUND AND METHODS: In the present study, we tested the hypothesis that peroxynitrite and subsequent activation of the nuclear enzyme poly(ADP-ribose) synthetase (PARS) play a role in the pathogenesis of multiple organ failure induced by peritoneal injection of zymosan in the rat. Animals were randomly divided into six groups (ten rats for each group). The first group was treated with ip administration of saline solution (0.9% NaCl) and served as the sham group. The second group was treated with ip administration of zymosan (500 mg/kg suspended in saline solution). In the third and fourth groups, rats received ip administration of 3-aminobenzamide (10 mg/kg) 1 and 6 hrs after zymosan or saline administration, respectively. In the fifth and sixth groups, rats received ip administration of nicotinamide (50 mg/kg) 1 and 6 hrs after zymosan or saline administration, respectively. After zymosan or saline injection, animals were monitored for 72 hrs to evaluate systemic toxicity (conjunctivitis, ruffled fur, diarrhea, and lethargy), loss of body weight, and mortality. RESULTS: A severe inflammatory response, characterized by peritoneal exudation, high plasma and peritoneal levels of nitrate/nitrite (the breakdown products of nitric oxide), and leukocyte infiltration into peritoneal exudate, was induced by zymosan administration. This inflammatory process coincided with the damage of lung, small intestine, and liver as assessed by histologic examination and by an increase of myeloperoxidase activity, which is indicative of neutrophil infiltration. Zymosan-treated rats showed signs of systemic illness, significant loss of body weight, and high mortality rates. Peritoneal administration of zymosan in the rat also induced a significant increase in the plasma levels of peroxynitrite as measured by the oxidation of the fluorescent dihydrorhodamine 123. Immunohistochemical examination demonstrated a marked increase in the immunoreactivity to nitrotyrosine, a specific "footprint" of peroxynitrite, in the lung of zymosan-shocked rats. In vivo treatment with ip administration of 3 aminobenzamide (10 mg/kg, 1 and 6 hrs after zymosan injection) or nicotinamide (50 mg/kg, 1 and 6 hrs after zymosan injection) significantly decreased mortality, inhibited the development of peritonitis, and reduced peroxynitrite formation. In addition, PARS inhibitors were effective in preventing the development of organ failure because tissue injury and neutrophil infiltration, by myeloperoxidase evaluation, were reduced in the lung, small intestine, and liver. CONCLUSIONS: In conclusion, the major findings of our study are that peroxynitrite and the consequent PARS activation exert a role in the development of multiple organ failure and that PARS inhibition is an effective anti inflammatory therapeutic tool. PMID- 10470760 TI - Protective effect of N-acetylcysteine on multiple organ failure induced by zymosan in the rat. AB - BACKGROUND AND METHODS: In the present study, we evaluated the effect of N acetylcysteine treatment in a nonseptic shock model induced by zymosan in the rat. Animals were randomly divided into eight groups (ten animals in each group). The first group was treated with ip administration of saline solution (0.90% NaCl) and served as the sham group. The second group was treated with ip administration of zymosan (500 mg/kg suspended in saline solution). In the third and fourth groups, rats received ip administration of N-acetylcysteine (40 mg/kg; 1 and 6 hrs after administration of zymosan or saline). In the fifth and sixth groups, rats received ip administration of N-acetylcysteine (20 mg/kg; 1 and 6 hrs after zymosan or saline administration). In the seventh and eighth groups, rats received ip administration of N-acetylcysteine (10 mg/kg; 1 and 6 hrs after zymosan or saline administration). After zymosan or saline injection, animals were monitored for the evaluation of systemic toxicity (conjunctivitis, ruffled fur, diarrhea, and lethargy), loss of body weight, and mortality for 72 hrs. Exudate formation, leukocyte infiltration, nitrate/nitrite production, lung and intestine myeloperoxidase activity and lipid peroxidation, and histologic examination were evaluated at 18 hrs after zymosan administration. RESULTS: Administration of zymosan in the rat induced acute peritonitis, as assessed by a marked increase in the leukocyte count in the exudate, as well as by an increase in the exudate nitrate/nitrite concentration. Lung and intestine myeloperoxidase activity and lipid peroxidation was significantly increased in zymosan-treated rats. This inflammatory process coincided with the damage of lung and small intestine. Peritoneal administration of zymosan in the rat also induced a significant increase in the plasma levels of nitrite and nitrate and stable metabolites of nitric oxide and in levels of peroxynitrite, as measured by the oxidation of the fluorescent dihydrorhodamine 123 at 18 hrs after zymosan challenge. Immunohistochemical examination demonstrated a marked increase in the immunoreactivity to nitrotyrosine, a specific "footprint" of peroxynitrite, in the lung of zymosan-shocked rats. Pretreatment of zymosan-shocked rats with ip administration of N-acetylcysteine (40, 20, and 10 mg/kg, 1 and 6 hrs after zymosan) prevented the development of peritonitis and reduced peroxynitrite formation in a dose-dependent manner. In addition, ip administration of N acetylcysteine (40 mg/kg, 1 and 6 hrs after zymosan) was effective in preventing the development of lung and intestine injury and neutrophil infiltration, as determined by myeloperoxidase evaluation. CONCLUSIONS: Taken together, the present results demonstrate that N-acetylcysteine exerts potent anti-inflammatory effects. PMID- 10470761 TI - Effects of mean airway pressure and tidal excursion on lung injury induced by mechanical ventilation in an isolated perfused rabbit lung model. AB - OBJECTIVE: To study the relative contributions of mean airway pressure (mPaw) and tidal excursion (V(T)) to ventilator-induced lung injury under constant perfusion conditions. DESIGN: Prospective, randomized study. SETTING: Experimental animal laboratory. SUBJECTS: Fifteen sets of isolated rabbit lungs. INTERVENTIONS: Rabbit lungs were perfused (constant flow, 500 mL/min; capillary pressure, 10 mm Hg) and randomized to be ventilated at identical peak transpulmonary pressure (pressure control ventilation [30 cm H2O and frequency of 20/min]) with three different ventilatory patterns that differed from each other by either mPaw or V(T): group A (low mPaw [13.4+/-0.2 cm H2O]/large V(T) [55+/-8 mL], n = 5); group B (high mPaw [21.2+/-0.2 cm H2O]/small V(T) [18+/-1 mL], n = 5); and group C (high mPaw [21.8+/-0.5 cm H2O]/large V(T) [53+/-5 mL], n = 5). MEASUREMENTS AND MAIN RESULTS: Continuous weight gain (edema formation), change in ultrafiltration coefficient (deltaKf, vascular permeability index), and histology (lung hemorrhage) were examined. In group A, deltaKf (0.08+/-0.08 g/min/cm H2O/100 g) was less than in group B (0.28+/-0.19 g/min/cm H2O/100 g) or group C (0.41+/-0.29 g/min/cm H2O/100 g) (p = .05). Group A experienced significantly less hemorrhage (histologic score, 5.4+/-2.2) than groups B (10.3+/-2.1) and C (11.1+/-3.0) (p < .05). A similar trend was observed for weight gain. In contrast to tidal excursion, mPaw was found to be a significant factor for lung hemorrhage and increased Kf (two-way analysis of variance; p < .05). Weight gain (r2 = .54, p = .04) and lung hemorrhage (r2 = .65, p = .01) correlated with the mean pulmonary artery pressure changes that resulted from the implementation of the ventilatory strategies. The difference between the changes in mPaw and mean pulmonary artery pressure linearly predicted deltaKf (p = .005 and .05, respectively, r2 = 0.73). CONCLUSIONS: Under these experimental conditions, mPaw contributes more than tidal excursion to lung hemorrhage and permeability alterations induced by mechanical ventilation. PMID- 10470762 TI - Myocardial effects of ventricular fibrillation in the isolated rat heart. AB - OBJECTIVE: Ventricular fibrillation (VF) is known to increase myocardial oxygen requirements and to alter coronary vascular physiology. However, the significance of these effects during cardiac arrest and resuscitation is not well understood. A model was developed in the isolated rat heart to investigate the myocardial effects of VF during a simulated episode of cardiac arrest and resuscitation. We hypothesized that VF would intensify the severity of myocardial ischemia and consequently accentuate postischemic myocardial dysfunction. DESIGN: Prospective and randomized. SETTING: Research laboratory. SUBJECTS: Twenty Sprague-Dawley rats. INTERVENTIONS: Hearts were harvested and perfused at a constant flow rate of 10 mL/min using a modified Krebs-Henseleit solution equilibrated with 95% oxygen and 5% CO2. In five hearts, VF was induced by a 0.05-mA current delivered to the right ventricular endocardium. The perfusate flow was then stopped for a 10-min interval and resumed at 20% of baseline flow for another 10 mins. After 20 mins of VF, the perfusate flow was returned to baseline and a sinus rhythm reestablished by epicardial electrical shocks. The studies were randomized and included three additional groups to control for the effects of ischemia without VF (n = 5), the effects of VF without ischemia (n = 5), and the stability of the preparation (n = 5). MEASUREMENTS AND MAIN RESULTS: Isovolumic indices of left ventricular function were obtained using a latex balloon advanced through the mitral valve and distended to an end-diastolic pressure of 10 mm Hg. The coronary effluent was collected from the right ventricular cavity. VF during myocardial ischemia was associated with a higher coronary effluent PCO2, increased coronary vascular resistance, and development of ischemic contracture as indicated by increases in left ventricular pressure from 9+/-3 to 33+/-6 mm Hg (p < .05). After defibrillation, contractility and relaxation rapidly returned to baseline values, whereas the isovolumic end-diastolic pressure remained elevated for 20 mins. These changes were much less prominent when ischemia was not accompanied by VF. CONCLUSIONS: These findings indicate that VF may adversely affect myocardial ischemia by hastening the development of ischemic contracture, increasing coronary vascular resistance, and favoring the development of diastolic pump failure early after resuscitation from cardiac arrest. PMID- 10470763 TI - Differential bronchodilatory effects of terbutaline, diltiazem, and aminophylline in canine intraparenchymal airways. AB - OBJECTIVES: Intraparenchymal airways are involved in air flow regulation. Relaxation of intraparenchymal airways to volatile anesthetics varied by topographic location. This study was conducted to determine whether other bronchodilators (terbutaline, diltiazem, and aminophylline) relax bronchiolus to a greater degree than bronchus, as seen with volatile anesthetics. DESIGN: In vitro, controlled, randomized study. SETTING: Animal research laboratory. SUBJECTS: Adult dogs (n = 9). INTERVENTIONS: Proximal (outer diameter, 4-6 mm) and distal (outer diameter, 0.8-1.5 mm) airway rings of dogs were contracted in tissue baths with the effective concentration of acetylcholine that produces half the maximum response. Airway relaxant dose-response curves were constructed to measure isometric tension after administration of terbutaline (concentration range, 10(-8) to 10(-4) M), diltiazem (concentration range, 3 x 10(-7) to 1 x 10( 4) M), and aminophylline (concentration range, 10(-7) to 10(-4) M). MEASUREMENTS AND MAIN RESULTS: All three bronchodilators caused relaxation of the proximal and distal airways. At the maximum dose, diltiazem (maximum relaxation, 95%+/-2% [proximal], 94%+/-6% [distal]; p > .05) was the most efficacious, followed by terbutaline (maximum relaxation, 72%+/-13% [proximal], 55%+/-9% [distal]; p < .05) and aminophylline (maximum relaxation, 32%+/-10% [proximal], 35%+/-18% [distal]; p > .05. At the concentrations tested, they were equally efficacious. No significant differences in relaxation between proximal and distal airways were noted with diltiazem or aminophylline in the entire dose range. However, terbutaline relaxed the distal airway more than the proximal airway in the entire dose range. CONCLUSIONS: The results demonstrate that only terbutaline showed a differential airway relaxant effect between proximal and distal airways, as seen with volatile anesthetics. PMID- 10470764 TI - Mild or moderate hypothermia but not increased oxygen breathing prolongs survival during lethal uncontrolled hemorrhagic shock in rats, with monitoring of visceral dysoxia. AB - OBJECTIVE: To test the hypotheses that during lethal uncontrolled hemorrhagic shock (UHS) in rats compared with normothermia and room air breathing: a) mild hypothermia would prolong survival time as well as moderate hypothermia; b) oxygen breathing would prolong survival further; and c) hypothermia and oxygen would mitigate visceral ischemia (dysoxia) during UHS. DESIGN: Prospective, randomized, controlled laboratory animal study. SETTING: Animal research facility. SUBJECTS: Male Sprague-Dawley rats. INTERVENTION: Fifty-four rats were lightly anesthetized with halothane during spontaneous breathing. UHS was induced by blood withdrawal of 3 mL/100 g over 15 mins, followed by 75% tail amputation with topical application of heparin. Five minutes after tail cut, rats were randomly divided into nine groups (6 rats each) with three rectal temperature levels (38 degrees C [100.4 degrees F; normothermia] vs. 34 degrees C [93.2 degrees F; mild hypothermia] vs. 30 degrees C [86 degrees F; moderate hypothermia]) by surface cooling; each with 3 FIO2 levels (0.25 vs. 0.5 vs. 1.0). Rats were observed without fluid resuscitation until death (apnea and pulselessness). Visceral ischemia was monitored by observing liver and gut surface PCO2. MEASUREMENTS AND MAIN RESULTS: Mean survival time, which was 51 mins in the control group with normothermia and FIO2 of 0.25, was more than doubled with hypothermia, to 119 mins in the combined mild hypothermia groups (p < .05) and to 132 mins in the combined moderate hypothermia groups (p < .05; NS for moderate vs. mild hypothermia). FIO2 had no statistically significant effect on survival time. Increases in visceral surface PCO2 correlated with hypotension (r2 = .22 for intestine and .40 for liver). Transiently, increased FIO2, not hypothermia, mitigated visceral ischemia. CONCLUSIONS: Both mild and moderate hypothermia prolonged survival time during untreated, lethal UHS in rats. Increased FIO2 had no effect on survival. The effects of hypothermia and increased FIO2 during UHS on viscera, the ability to be resuscitated, and outcome should be explored further. PMID- 10470765 TI - Intraosseous vasopressin improves coronary perfusion pressure rapidly during cardiopulmonary resuscitation in pigs. AB - OBJECTIVE: Intravenous administration of vasopressin during cardiopulmonary resuscitation (CPR) may be more effective than optimal doses of epinephrine. The main purpose of this study was to determine whether intraosseous vasopressin achieves serum drug levels comparable with intravenous doses during CPR and, additionally, to evaluate the effects of intraosseous vasopressin during CPR. DESIGN: Prospective, randomized laboratory investigation using an established porcine model with instrumentation for measurement of hemodynamic variables, blood gases, and return of spontaneous circulation. SETTING: University hospital laboratory. SUBJECTS: Twelve domestic pigs. INTERVENTIONS: After 4 mins of untreated ventricular fibrillation and 3 mins of CPR, 12 pigs were randomized to be treated with intravenous administration of vasopressin (0.8 unit/kg vasopressin; n = 6) or intraosseous vasopressin (0.8 unit/kg vasopressin; n = 6). Defibrillation was performed 5 mins after drug administration to attempt the return of spontaneous circulation. MEASUREMENTS AND MAIN RESULTS: At both 90 secs and 5 mins after drug administration, intravenous and intraosseous administration of vasopressin resulted in comparable mean (+/-SEM) coronary perfusion pressure (43+/-4 vs. 44+/-3 and 30+/-2 vs. 37+/-2 mm Hg, respectively) and vasopressin plasma concentrations (13,706+/-1,857 vs. 16,166+/-3,114 pg/mL and 10,372+/-883 vs. 8246+/-2211 pg/mL, respectively). All animals in both groups were successfully resuscitated; pigs that received intraosseous vasopressin had a significantly higher (p < .05) mean arterial (92+/-6 vs. 129+/-12 mm Hg) and coronary perfusion pressure (84+/-11 vs. 119+/-11 mm Hg) at 5 mins of return of spontaneous circulation. CONCLUSIONS: Intraosseous vasopressin resulted in comparable vasopressin plasma levels, hemodynamic variables, and return of spontaneous circulation rates as did intravenous vasopressin. Intraosseous vasopressin may be an alternative for vasopressor administration during CPR, when intravenous access is delayed or not available. PMID- 10470766 TI - Elimination of methohexitone after long-term, high-dose infusion in patients with critically elevated intracranial pressure. AB - OBJECTIVE: To determine the plasma elimination of methohexitone in patients with critically elevated intracranial pressure (ICP) who received the drug in high doses for several days. DESIGN: Drug-monitoring study. SETTING: Intensive care unit at a university hospital. PATIENTS: Twelve intensive care unit patients with brain injuries who received methohexitone as a final therapeutic approach after routine therapy had proved to be insufficient in controlling critically elevated ICP. MEASUREMENTS AND MAIN RESULTS: Plasma samples were taken during methohexitone infusion, before cessation, and in distinct, short increments after discontinuation of the infusion. Methohexitone was determined in plasma by reverse-phase high-pressure liquid chromatography and photometric detection. The median duration of infusion of methohexitone was 137 hrs (minimum, 27 hrs; maximum, 445 hrs), with a median infusion rate of 62.5 microg/kg/min (minimum, 22.5 microg/kg/min; maximum, 116.2 microg/kg/min). Plasma concentrations of methohexitone at burst suppression under concomitant analgesic sedation ranged from 1.6 to 17.3 microg/mL (median, 4.7 microg/mL). After cessation of methohexitone infusion, the decline of plasma concentrations followed a biexponential function. Clearance rates, volume of distribution at steady state, context-sensitive half-time, and initial and terminal elimination half-times were calculated. Pharmacokinetic data showed remarkable interindividual variability that could not be correlated to the infusion rate, to the duration of the infusion, or to obvious differences in physiology or the disease states of these patients. Even in patients with high plasma concentrations who received the drug for a considerable length of time, the initial decline in plasma concentration was exponential, indicating redistribution. CONCLUSIONS: We conclude that the elimination kinetics of methohexitone after long-term, high-dose infusion in critically ill patients with brain injuries may favor the use of methohexitone over thiopentone for controlling critically elevated ICP by allowing for a more timely neurologic examination after cessation. PMID- 10470767 TI - Unmeasured anions identified by the Fencl-Stewart method predict mortality better than base excess, anion gap, and lactate in patients in the pediatric intensive care unit. AB - OBJECTIVES: This study was undertaken to compare three methods for the identification of unmeasured anions in pediatric patients with critical illness. We compared the base excess (BE) and anion gap (AG) methods with the less commonly used Fencl-Stewart strong ion method of calculating BE caused by unmeasured anions (BEua). We measured the relationship of unmeasured anions identified by the three methods to serum lactate concentrations and to mortality. DESIGN: Retrospective cohort study. SETTING: Tertiary care pediatric intensive care unit in an academic pediatric hospital. PATIENTS: The study population included 255 patients in the pediatric intensive care unit who had simultaneous measurements of arterial blood gases, electrolytes, and albumin during the period of July 1995 to December 1996. Sixty-six of the 255 patients had a simultaneous measurement of serum lactate. MEASUREMENTS AND MAIN RESULTS: The BEua was calculated using the Fencl-Stewart method. The AG was defined as (sodium plus potassium) - (chloride plus total carbon dioxide). BE was calculated from the standard bicarbonate, which is derived from the Henderson-Hasselbalch equation and reported on the blood gas analysis. A BE or BEua value of < or =-5 mEq/L or an AG > or =17 mEq/L was defined as a clinically significant presence of unmeasured anions. A lactate level of > or =45 mg/dL was defined as being abnormally elevated for this study. The presence of unmeasured anions identified by significantly abnormal BEua was poorly identified by BE or AG. Of the 255 patients included in the study, 67 (26%) had a different interpretation of acid base balance when the Fencl method was used compared with when BE and AG were used. Plasma lactate concentration correlated better with BEua (r2 = .55; p = .0001) than with AG (r2 = .41; p = .0005) or BE (r2 = .27; p = .025). Mortality was more strongly related to BEua < or =-5 mEq/L (relative risk of death = 10.25; p = .002) than to lactate > or =45 mg/dL (relative risk of death = 2.35; p = .04). In logistic regression analysis, mortality was more strongly associated with BEua (area under the receiver operating characteristic curve = 0.79; p = .0002) than lactate (receiver operating characteristic curve area = 0.63; p = .05), BE (receiver operating characteristic curve area = 0.53; p = .32), or AG (receiver operating characteristic curve area = 0.64; p = .08) in this patient sample. CONCLUSIONS: Critically ill patients with normal BE and normal AG frequently have elevated unmeasured anions detectable by BEua. The Fencl-Stewart method is better than BE and similar to AG in identifying patients with high lactate levels. Elevated unmeasured anions identified by the Fencl-Stewart method were more strongly associated with mortality than with BE, AG, or lactate in this patient sample. PMID- 10470768 TI - Whatever happened to Reye's syndrome? Did it ever really exist? AB - OBJECTIVE: Reye's syndrome (RS) appeared suddenly in the 1950s and disappeared almost as quickly in the late 1980s. A number of metabolic disorders were discovered in the 1980s that could completely mimic RS. This study was undertaken to reassess the original diagnosis of RS in light of newly described metabolic disorders. INTERVENTION: The medical records of 26 patients who had survived RS and were originally studied in Australia in the 1980s were reexamined 10 yrs later, and families were interviewed to ascertain if the diagnosis had changed. The 49 original patients with RS from Australia were also reanalyzed using more precise diagnostic criteria for RS to ascertain how many of the patients would continue to fit the more precise diagnosis of RS. MEASUREMENTS AND MAIN RESULTS: Of 26 original patients with RS who had survived, 18 (69%) were subsequently diagnosed as having other diseases, most commonly inborn errors of metabolism. The most commonly diagnosed metabolic disorder was medium-chain acyl-coenzyme-A dehydrogenase deficiency. Of the 18 patients rediagnosed with diseases other than RS, 15 (83%) are now known to have metabolic disorders. By using more precise diagnostic criteria for RS, none of the original 49 patients with RS could be diagnosed as having certain RS. Only six patients had probable RS, two patients had possible RS, 23 patients had unlikely RS, and 18 patients were excluded as RS cases. CONCLUSION: With better diagnostic techniques and criteria, most patients originally diagnosed with RS are now known to have metabolic disorders. The disappearance of RS was probably related to the discovery and ability to diagnose inborn errors of metabolism that mimicked RS clinically, biochemically, and pathologically. PMID- 10470769 TI - Financial impact of elimination of routine chest radiographs in a pediatric intensive care unit. AB - OBJECTIVE: To determine the change in chest radiograph use if each chest radiograph requires a separate order and clinical indication. DESIGN: Prospective, nonrandomized, controlled design with an intervention. SETTING: The pediatric intensive care unit (PICU) at Primary Children's Medical Center, Salt Lake City, UT. PATIENTS: The study comprised 3,727 PICU patients treated between 1992 and 1996. INTERVENTIONS: A change in ordering practice: There will be no standing orders for routine daily morning chest radiographs. Each radiograph requires a written order and a clinical indication. MEASUREMENTS AND MAIN RESULTS: During a 29-month control phase when routine daily chest radiographs were obtained for all intubated patients, 1.026 chest radiographs per patient day were performed. After the intervention, the ratio dropped to 0.653 chest radiographs per patient day, a decrease of 36.4%. This resulted in a (projected) variable cost savings of $45,476. Data were also collected for quality assurance purposes. CONCLUSIONS: These results demonstrate the impact of an evaluation and subsequent change in radiology ordering practice in our PICU. The change resulted in decreased variability in ordering practice, fewer chest radiographs per patient, and an accompanying cost savings to our patients and payors. PMID- 10470770 TI - Distribution and pathophysiology of acute lobar collapse in the pediatric intensive care unit. AB - OBJECTIVE: The high incidence of lower lobe collapse in adult intensive care patients is well described. We aimed to document the incidence and distribution of acute lobar collapse in the pediatric intensive care setting. The influence of anatomical and pathophysiological differences between the adult and pediatric respiratory tract will be considered. DESIGN: Retrospective review of chest radiograph series. SETTING: Tertiary referral center for pediatric intensive care and the Department of Diagnostic Radiology in a large teaching hospital in England. PATIENTS: Cohort of 160 patients receiving intensive care during a 2-yr period (age range, 6 days-18 yrs; median, 23 months). INTERVENTIONS: None MEASUREMENTS AND MAIN RESULTS: Twenty-four of 160 children (15%) developed acute lobar collapse during their intensive care unit admission. Isolated right upper lobe collapse occurred in 14 patients, right upper lobe in association with one or more other lobes in five patients, and lobar collapse other than the right upper lobe in five patients. The development of lobar collapse and, in particular, right upper lobe collapse was associated with a lower median age (no collapse, 26 months; lobar collapse, 8 months; right upper lobe collapse, 4 months). Lobar collapse was significantly associated with the requirement for mechanical ventilation during admission (chi-square, 12.18; p = .005). It was observed in association with both high and low endotracheal tube positions. CONCLUSION: The predominance of upper lobe and, in particular, right upper lobe collapse observed in pediatric intensive care patients contrasts with the high incidence of lower lobe collapse in their adult counterparts. Multiple interrelated factors are likely to be contributory and include the following: a) anatomical and physiological differences between adults and children; b) the pathophysiology of childhood respiratory disease; c) more critical positioning of endotracheal tubes in younger patients and their movement with patient positioning. PMID- 10470771 TI - Development of a percutaneous fiberoptic hepatic venous localization catheter. AB - OBJECTIVE: To develop a liver-specific biosensor system/catheter assembly that can be used to localize and cannulate the hepatic venous system without the need for fluoroscopic imaging. This would permit the bedside placement of a hepatic venous catheter for monitoring purposes without radiographic guidance. DESIGN: Experimental, in vitro. STUDY SETTING: Experimental laboratory at a university center. SUBJECT: This was a simulation study to evaluate the ability of a cardiovascular monitoring catheter mounted with a liver-specific biosensor to anatomically identify a side arm tributary. The experimental system used for this study mimics the hepatic vein draining into the inferior vena cava and allows its localization without the need for assisted imaging. The biosensor design and catheter/sensor assembly function were studied in this in vitro model. INTERVENTIONS: A liver-specific biosensor was developed by housing a homogeneous affinity fluorescence assay system sensitive to galactose in a microdialysis hollow fiber receptacle. A polyvinyl chloride tube containing a side arm was constructed to mimic the confluence of a venous tributary (i.e., the hepatic vein) with a major vascular channel (i.e., the vena cava). In this simulation, the side arm was continuously perfused with a liver-sensitive analyte (galactose) and the main channel was perfused with galactose-free buffer. A cardiovascular catheter containing a fiberoptic waveguide mounted with a galactose-sensitive fluorescent probe was advanced along the main conduit to assess its ability to identify the location of the galactose side arm infusion site. MEASUREMENTS AND MAIN RESULTS: The response of the fiberoptic sensor to different galactose concentrations was assessed and found to be almost linear over the concentration range of 0 to 2 mM, which encompasses the expected utilization range of this system. The variability in identifying the galactose infusion point (simulated hepatic vein) in a 15-cm conduit was 1.7 to 2.8 mm, or 1.1% to 1.9%. CONCLUSIONS: The construction of a catheter/sensor system with the ability to provide accurate spatial/anatomical localization data for the hepatic venous system is feasible. This assembly will eliminate the need for ancillary imaging systems for catheter/sensor delivery to an individual organ system and potentially can be positioned at the bedside in a fashion similar to the pulmonary artery flotation catheter. PMID- 10470772 TI - Effect of nitric oxide, perfluorocarbon, and heliox on minute volume measurement and ventilator volumes delivered. AB - OBJECTIVE: To determine the effect of heliox, nitric oxide (NO), and perfluorocarbon on differential pressure pneumotachometer characteristics and to determine the effect of heliox on volumes delivered by the Siemens S900C (S900C), and Servo Ventilator 300 (SV300) ventilators. DESIGN: Prospective, laboratory study. SETTING: Pulmonary laboratory of a tertiary care, nonprofit children's hospital. APPARATUS: SV300, S900C ventilator, differential pressure pneumotachometer. INTERVENTIONS: Dual pneumotachometers were connected in series to a 0.5-L calibration syringe and a 1-L anesthesia bag creating a closed system. Calibration of the pneumotachometers was done in room air at ambient temperature with 100 strokes. Accepted accuracy of measured volumes is within 0.5%. Flow conductance curves were constructed using 100 strokes each for heliox (70:30 mixture), NO, and perfluorocarbon. Expired gases of room air and a 70:30 mixture of heliox from the above ventilators were collected into a nondiffusing gas collection bag, and the volume was measured in a chain-compensated gasometer. Ten sets of 500-mL breaths (20 breaths each set) and 100-mL breaths (40 breaths each set) were collected. The paired Student's t-test was used to detect significant differences in measured volumes, with significance defined as p < .01. MEASUREMENTS AND MAIN RESULTS: Volumes measured with the pneumotachometer using 25 ppm of NO, 50 ppm of NO, and perfluorocarbon were within +0.25%, -0.7%, and +0.4%, respectively (p = .155, p = .001, p = .06). Heliox decreased the conductance of the pneumotachometer, thereby increasing the measured volume by 15% (p < .001). However, heliox did not affect its linearity. Heliox had no affect on volumes delivered by the S900C. However, the SV300 delivered 7.9% less volume of heliox at a set tidal volume of 500 mL and 10.8% less at a set tidal volume of 100 mL. CONCLUSIONS: A 70:30 mixture of heliox caused a significantly overestimated gas volume measured and, therefore, an underestimated gas volume delivered by SV300. NO at 25 ppm and perfluorocarbon did not interfere with the accuracy of a differential pressure pneumotachometer. However, at 50 ppm, NO caused a difference in measured gas volume that was statistically, but not clinically, significant. Application of pneumotachometers in critically ill children receiving heliox requires recalibration. Heliox did not affect volumes delivered with the S900C ventilator. Although volumes delivered with the SV300 were significantly reduced by heliox, the difference can be corrected easily by increasing minute ventilation until expired tidal volume equals desired tidal volume. PMID- 10470773 TI - Clinical gram-positive sepsis: does it fundamentally differ from gram-negative bacterial sepsis? AB - OBJECTIVE: To review the basic differences between gram-positive and gram negative sepsis and to assess the effect of these differences on current and future therapeutic strategies for sepsis. DESIGN: Literature review of the past 30 yrs of laboratory and clinical reports that analyze the microbial aspects of sepsis and the immunologic response to systemic infection. RESULTS: The increasing prevalence of sepsis from gram-positive bacterial pathogens necessitates reevaluation of many of the basic assumptions about the molecular pathogenesis of septic shock. It has been assumed that the initiation of the systemic inflammatory response with activation of the proinflammatory cytokine networks and other mediators results in a similar pathophysiologic process, regardless of the causative microbic pathogen. Yet, there is increasing experimental evidence that fundamental differences exist in the host response to gram-positive bacterial pathogens compared with the host response to gram negative organisms. Systemic immune activation during sepsis may promote the clearance of the microbic pathogen; however, generalized inflammation also contributes to the pathogenesis of septic shock. The balance between these beneficial and deleterious effects may differ between gram-positive and gram negative pathogens. CONCLUSIONS: Results of antimediator therapies in clinical trials in septic shock are inconclusive but suggest that the response may differ, depending on the type of microbic pathogen. The immune-mediated pathophysiologic mechanisms that underlie gram-positive sepsis and the potential interactions between the infecting microorganism and efficacy of anticytokine therapies require further investigation. Treatment strategies that explain the causative organism may be necessary for optimal use of immunoadjuvants in the future. PMID- 10470774 TI - Percutaneous or surgical tracheostomy: a meta-analysis. AB - OBJECTIVE: To compare percutaneous with surgical tracheostomy using a meta analysis of studies published from 1960 to 1996. DATA SOURCES: Publications obtained through a MEDLINE database search with a Boolean combination (tracheostomy or tracheotomy) and complications, with constraints for human studies and English language. STUDY SELECTION: Publications addressing all peri- and postoperative complications. Studies limited to specific tracheostomy complications or containing insufficient details were excluded. Two authors independently selected the publications. DATA EXTRACTION: A list of relevant surgical variables and complications was compiled. Complications were divided into peri- and postoperative groups and further subclassified into severe, intermediate, and minor groups. Because most studies of percutaneous tracheostomy were published after 1985, surgical tracheostomy studies were divided into two periods: 1960 to 1984 and 1985 to 1996. The articles were analyzed independently by three investigators, and rare discrepancies were resolved through discussion and data reexamination. DATA SYNTHESIS: Earlier surgical tracheostomy studies (n = 17; patients, 4185) have the highest rates of both peri- (8.5%) and postoperative (33%) complications. Comparison of recent surgical (n = 21; patients, 3512) and percutaneous (n = 27; patients, 1817) tracheostomy trials shows that perioperative complications are more frequent with the percutaneous technique (10% vs. 3%), whereas postoperative complications occur more often with surgical tracheotomy (10% vs. 7%). The bulk of the differences is in minor complications, except perioperative death (0.44% vs. 0.03%) and serious cardiorespiratory events (0.33% vs. 0.06%), which were higher with the percutaneous technique. Heterogeneity analysis of complication rates shows higher heterogeneity in older and surgical trials. CONCLUSIONS: Percutaneous tracheostomy is associated with a higher prevalence of perioperative complications and, especially, perioperative deaths and cardiorespiratory arrests. Postoperative complication rates are higher with surgical tracheostomy. PMID- 10470775 TI - Forgoing life support in western European intensive care units: the results of an ethical questionnaire. AB - OBJECTIVE: To determine current views of European intensive care physicians regarding end-of-life decisions. DESIGN: A questionnaire was sent to all physician members of the European Society of Intensive Care Medicine. All questionnaires were anonymous. RESULTS: A total of 504 completed questionnaires from 16 western European countries were analyzed. Eighty-seven percent of the respondents were male. Forty-six percent of respondents said that intensive care unit admissions were generally or commonly affected by bed shortages, particularly in the south. Nevertheless, 73% of units frequently admit patients with no hope of survival, although only 33% of respondents felt that such patients should be admitted. Eighty percent of respondents felt that written do not-resuscitate orders should be applied, but only 58% did so, with a wide variation according to country (from 8% in Italy to 91% in The Netherlands). Ninety-three percent of physicians sometimes withhold treatment from patients with no hope of a meaningful life, but withdrawal of treatment is less common. Forty percent of respondents said that they would deliberately administer large doses of drugs to such patients until death ensued. Forty-nine percent of respondents involved staff, patients, and family in end-of-life decisions. Forty five percent of respondents felt that an ethics consultation was useful in such situations. Physicians in the countries of southern Europe were less likely than those in the north to apply do-not-resuscitate orders, withhold treatment, and discuss such issues with the patients. However, they were more likely to value the opinion of an ethics consultant. CONCLUSIONS: Intensive care unit admissions are frequently limited by the availability of beds across Europe, particularly in the south and in the United Kingdom, yet 73% of intensivists still admit patients with no hope of survival. When treating patients with no hope of survival, 40% of intensivists will deliberately administer large doses of drugs until death ensues. There are interesting differences between what a physician actually does and what he or she believes should be done with regard to various ethical questions. Important differences in attitudes also exist between European countries. PMID- 10470776 TI - Recombinant human growth hormone for reconditioning of respiratory muscle after lung volume reduction surgery. AB - OBJECTIVE: To investigate the effects of recombinant human growth hormone (rHGH) as a "rescue treatment" in an end-stage chronic obstructive pulmonary disease patient after prolonged weaning failure. DESIGN: Descriptive case report. SETTING: Fifteen-bed intensive care unit in a university hospital. PATIENT: A 62 year-old man with end-stage chronic obstructive pulmonary disease and pulmonary emphysema after lung reduction surgery and prolonged weaning failure after long term mechanical ventilation. INTERVENTIONS: After 42 days of unsuccessful weaning from the respirator, rHGH (27 IU/day, 0.3 IU/kg body weight/day) was administered for 20 days through a subcutaneous injection in addition to standard intensive care. MEASUREMENTS AND MAIN RESULTS: In addition to daily routine laboratory studies, the visceral proteins prealbumin, retinol-binding protein, and transferrin, and nitrogen balance were measured twice a week, as were the thyroid hormones triiodothyronine, thyroxine, and thyroid-stimulating hormone, plasma insulin levels, and the insulin-like growth factor (IGF)-1 binding proteins IGF BP1 and IGF-BP3. IGF-1 was measured from day 1 to day 4 of rHGH administration. Nutritional support was guided by indirect calorimetry. Additionally, weaning variables such as peak expiratory flow rate and expiratory tidal volume were measured noninvasively. T-piece weaning trials were carried out daily until respiratory muscle fatigue occurred. IGF-1 increased in response to rHGH stimulation, from 103 to 230 microg/mL, within 4 days. The carrier protein IGF BP3 increased from 126 to 283 mg/L at the end of the study period, and the inhibiting IGF-BP1 decreased initially from 19 to 14 mg/L and then increased until the end of the study to 31 mg/L. Nitrogen balance increased initially from 4.6 to 13.6 g/24 hrs and thereafter decreased until the end of rHGH treatment to 8.3 g/24 hrs. Resting energy expenditure increased from 1800 to 2300 kcal/24 hrs. Peak expiratory flow rate increased from 0.69 to 0.88 L/sec. The expiratory tidal volume showed a slight increase during the study period during the daily decrease of pressure support on the ventilator setting. Respiratory muscular strength increased beginning 10 days after rHGH therapy was started. From this point, T piece weaning trials could be prolonged almost daily. The patient was extubated successfully on postoperative day 75. CONCLUSIONS: This case report shows that after a prolonged catabolic state and long-term mechanical ventilation, administration of rHGH not only enhances the response of protein metabolism but improves respiratory muscular strength. Therefore, it may reduce the duration of mechanical ventilation in selected patients. PMID- 10470777 TI - Treatment of cachexia with recombinant growth hormone in a patient before lung transplantation: a case report. AB - OBJECTIVES: To describe the effects of recombinant human growth hormone (rhGH) in a severely malnourished patient before lung transplantation. DESIGN: Case study. SETTING: Intensive care unit. PATIENTS: A 38-yr-old severely malnourished (body mass index, 15.1 kg/m2) woman (receiving prednisone) with bronchiolitis obliterans evolving during 10 yrs presented with end-stage lung disease and required continuous noninvasive mechanical ventilation. INTERVENTIONS: Two courses of 35 days of 16 IU/day (0.42 IU/kg/day) rhGH administered subcutaneously, with an interruption of 5 wks between the two courses of rhGH. MEASUREMENTS AND MAIN RESULTS: Weight gain of 14.7% and 12.8% fat-free mass, as measured by 50-kHz bioelectrical impedance analysis, during treatment during a 3.5-month period. Nitrogen excretion decreased from 23.7 g/day before treatment to 8.0 g/day while receiving rhGH. Improvement of pulmonary function was also noted and allowed discharge of the patient from the hospital after the second course of rhGH. She underwent successful lung transplantation 2 months later and reached 48.8 kg of body weight 6 months later. CONCLUSIONS: rhGH treatment is a possible strategy that could be used with malnourished patients who are awaiting lung transplantation to improve the nutritional status and respiratory muscle function to prevent recurring respiratory infection and postoperative complications favored by malnutrition and possibly to decrease the length of hospital stay. PMID- 10470779 TI - Dietary lipids: ancestral ligands and regulators of cell signaling pathways. PMID- 10470778 TI - Traumatic macroglossia: a life-threatening complication. AB - OBJECTIVE: To describe the use of muscle relaxants and a bite raiser to avoid continued tongue trauma. DESIGN: Case report. SETTING: A tertiary general intensive care unit (ICU). INTERVENTIONS: Muscle relaxation and bite raiser. MAIN RESULTS: Muscle relaxation and a bite raiser were used in a 17-yr-old male with traumatic macroglossia, which allowed for rapid resolution of edema and prevented additional trauma to the tongue. CONCLUSION: Early use of a bite raiser together with muscle relaxants allows for more rapid solution of edema and prevention of additional trauma to the tongue in patients with traumatic macroglossia. PMID- 10470780 TI - Bacteremia in the intensive care unit: a predicament propagated by progress. PMID- 10470781 TI - Intestinal mucosal permeability: mechanisms and implications for treatment. PMID- 10470782 TI - Enteral nutrition: should we feed the stomach? PMID- 10470783 TI - Chest wall elastance and acute respiratory failure. PMID- 10470784 TI - Bioimpedance and the estimation of net fluid balance in critical care patients: problems and possibilities. PMID- 10470785 TI - Clinical trials in acute respiratory distress syndrome: what is ARDS? PMID- 10470786 TI - National intensive care unit datasets: lost at sea without a compass? PMID- 10470787 TI - Tidal volume, positive end-expiratory pressure, and mortality in acute respiratory distress syndrome. PMID- 10470788 TI - Bispectral Index: better information for sedation in the intensive care unit? PMID- 10470789 TI - Cardiac output minute to minute with a standard pulmonary artery catheter, without fluid overload? PMID- 10470790 TI - Deciphering the oxyradical inflammation Rosetta stone: O2*-NO*, OONO-, polymorphonuclear neutrophils, poly(ADP-ribose) synthetase, systemic inflammatory response syndrome, and multiple organ dysfunction syndrome. PMID- 10470791 TI - Ventilator-induced lung injury: more studies, more questions. PMID- 10470792 TI - Survival from ventricular fibrillation. PMID- 10470793 TI - Unmeasured anions: deja vu all over again? PMID- 10470794 TI - Reye's syndrome: hold the obituary. PMID- 10470795 TI - To chest x-rays and beyond!! PMID- 10470796 TI - Acute lobar collapse: adults and infants differ in important ways. PMID- 10470797 TI - Why, when, and how to insert a hepatic vein catheter in critically ill patients. PMID- 10470798 TI - Therapy of sepsis: why have we made such little progress? PMID- 10470799 TI - Percutaneous versus surgical tracheostomy: procedure of choice or choice of procedure. PMID- 10470800 TI - European life-support questionnaire. PMID- 10470801 TI - Assessing new technology, another chapter: lung volume reduction surgery/recombinant human growth hormone. PMID- 10470802 TI - Traumatic macroglossia: a new approach. PMID- 10470803 TI - In vitro activity of rifampin-minocyclin coating to Candida albicans. PMID- 10470804 TI - Another alternative for cardiac output measurement--pulse dye-densitometry. PMID- 10470805 TI - Gastrointestinal PiCO2 monitoring. PMID- 10470806 TI - Is heat shock protein-72 responsible for nitric oxide's ability to block stress induced apoptosis? PMID- 10470807 TI - Outcome after complex neurosurgery: the caregiver's burden is forgotten. AB - OBJECT: The goals of this study were twofold: 1) to determine outcome, including quality of life, in patients who have undergone surgery for petroclival meningioma in which a standard skull base approach was used; and 2) to assess the impact of the patients' surgical treatment on their caregivers. METHODS: Seventeen patients (13 women and four men ranging in age from 29 to 63 years) who underwent a transpetrosal approach for a petroclival meningioma during a 5-year period were prospectively included in this study. Pre- and postoperative data including adverse events were noted. The patients were assessed at 3, 6, and 12 months postoperatively, and annually thereafter, and they completed a postoperative SF-36 questionnaire. In addition, each patient's caregiver was interviewed to determine the effect of the patient's illness on the caregiver's life and responsibilities. Twenty-two operations were performed. A new permanent neurological deficit developed in five patients and in eight a temporary deficit or exacerbation of existing deficits occurred. Two patients underwent surgery to create a facial-hypoglossal nerve communication; five required a temporary percutaneous gastrostomy and/or tracheostomy; three required a shunt; and one underwent successful squint surgery. At 1 year postoperatively 13 patients had made a good or moderate recovery, three were severely disabled, and one had died- outcomes in keeping with other studies. By contrast, responses to the SF-36 questionnaire showed that, in all eight of its categories, between 43% and 75% of surviving patients were functioning below accepted norms. Fifty-six percent of caregivers experienced a major change in lifestyle and 38% experienced a major change with respect to their work. CONCLUSIONS: After transpetrosal excision of a petroclival meningioma, the quality of life for the patient is worse than that indicated in surgeons' reported results. The impact on the patient's caregiver is profound-a burden perhaps not fully appreciated by the surgeon. PMID- 10470808 TI - Natural history of asymptomatic colloid cysts of the third ventricle. AB - OBJECT: To determine the natural history of colloid cysts of the third ventricle in patients in whom the cysts were incidentally discovered, the authors retrospectively reviewed cases observed during the modern neuroimaging era (1974 1998). METHODS: During this 25-year interval, 162 patients with colloid cysts were examined and cared for at our center. Sixty-eight patients (42%) were thought to be asymptomatic with regard to their colloid cyst and observation with serial neuroimaging was recommended. The mean patient age was 57 years at the time of diagnosis (range 7-88 years) and the mean cyst size was 8 mm (range 4-18 mm). Computerized tomography scanning revealed a hyperdense cyst in 49 (84%) of 58 patients. Three patients were excluded from the study because they died of unrelated causes within 6 months of scanning and seven patients were lost to follow-up review. Clinical follow-up evaluation was available at a mean of 79 months (range 7-268 months) in the remaining 58 patients. The numbers of patients who participated in follow-up review at 2, 5, and 10 years after diagnosis were 40, 28, and 14, respectively. The incidences of symptomatic progression related to the cyst were 0%, 0%, and 8% at 2, 5, and 10 years, respectively. No patient died suddenly during the follow-up interval. Two (6%) of 34 patients in whom follow-up imaging was performed either exhibited cyst growth (one patient) or experienced hydrocephalus (one patient) at a mean of 41 months after diagnosis (range 4-160 months). CONCLUSIONS: Patients in whom asymptomatic colloid cysts are diagnosed can be cared for safely with observation and serial neuroimaging. If a patient becomes symptomatic, the cyst enlarges, or hydrocephalus develops, prompt neurosurgical intervention is necessary to prevent the occurrence of neurological decline from these benign tumors. PMID- 10470809 TI - Pineal epidermoid cysts: diagnosis and management. AB - OBJECT: The results of surgical treatment of epidermoid cysts of the pineal region in six cases are presented. METHODS: Six patients with pineal epidermoid cysts underwent surgery at the Institute of Neurosurgery "N. N. Burdenko," in Moscow, during the period 1976 to 1995. The duration of the patients' preadmission clinical history varied from 6 months to 2 years (average 1.4 years). Headache, diplopia, and vertigo were the most frequently occurring symptoms. Neurological examination demonstrated papilledema, impaired pupillary reaction, ataxia, and long-pathways deficit; Parinaud's syndrome was found in only one case. Computerized tomography and magnetic resonance imaging constituted the primary diagnostic tools. Surgery was performed using either an infratentorial-supracerebellar approach (two cases) or an occipital transtentorial approach (four cases). CONCLUSIONS: Surgical results can be excellent if a removal, as extensive as possible, is performed using either the supracerebellar or occipital-transtentorial approach. PMID- 10470810 TI - Predicting the probability of meningioma recurrence based on the quantity of peritumoral brain edema on computerized tomography scanning. AB - OBJECT: The goal of this study was to determine whether the quantity of peritumoral brain edema displayed on computerized tomography (CT) scanning could be correlated with brain invasion and subsequent recurrence of meningiomas. METHODS: One hundred thirty-five patients who underwent resection of intracranial meningiomas at the Ottawa Civic Hospital were followed during the period 1980 to 1998. A complete resection was defined as one in which tumor, invaded bone, and involved dura were removed. Tumors were examined microscopically for evidence of brain invasion. The mean follow-up period was 9 +/- 4 years (standard deviation [SD]) and the mean time to recurrence was 5 +/- 4 years (SD). The authors used a simple grading system based on the average thickness (in centimeters) of edema seen on an axial CT slice showing the most tumor. Edema grade was linearly related to edema volume determined by digitizing the scans (r = 0.96; 29 cases). The chance of brain invasion increased by 20% for each centimeter of edema (r(s) = 1, p < 0.0001; 124 cases). The presence of brain invasion was predictive of recurrence after complete resection with an accuracy of 83%, a sensitivity of 89%, and a specificity of 82%. The chance of recurrence within 10 years after complete resection was given by the equation: percentage chance of recurrence = (centimeter of edema)3 x 0.7, which can be used to predict the chance of recurrence based on findings on CT scans (r(s) = 1, p < 0.0001; 86 patients). Statistical significance was confirmed using Kaplan-Meier and univariate and multivariate analyses. Completeness of resection was the most powerful predictor of recurrence (p < 0.00001, r = 0.6), followed by edema grade and brain invasion (both p = 0.02, r = 0.1). Patient age and gender and tumor location, size, and histological subtype were nonsignificant factors. CONCLUSIONS: Brain invasion causes peritumoral edema. Invaded brain tissue is also the source of residual cells in cases of tumor recurrence after gross-total resection. PMID- 10470811 TI - Correlation of the relationships of brain-tumor interfaces, magnetic resonance imaging, and angiographic findings to predict cleavage of meningiomas. AB - OBJECT: The authors examined the relationships of brain-tumor interfaces, specific magnetic resonance (MR) imaging features, and angiographic findings in meningiomas to predict tumor cleavage and difficulty of resection. METHODS: Magnetic resonance imaging studies, angiographic data, operative reports, clinical data, and histopathological findings were examined retrospectively in this series, which included 126 patients with intracranial meningiomas who underwent operations in which microsurgical techniques were used. The authors have identified three kinds of brain-tumor interfaces characterized by various difficulties in microsurgical dissection: smooth type, intermediate type, and invasive type. The signal intensity on T1-weighted MR images was very similar regardless of the type of brain-tumor interface (p > 0.1). However, on T2 weighted images the different interfaces seemed to correlate very precisely with the signal intensity and the amount of peritumoral edema (p < 0.01), allowing the prediction of microsurgical effort required during surgery. On angiographic studies, the pial-cortical arterial supply was seen to participate almost equally with the meningeal-dural arterial supply in vascularizing the tumor in 57.9% of patients. Meningiomas demonstrating hypervascularization on angiography, particularly those fed by the pial-cortical arteries, exhibited significantly more severe edema compared with those supplied only from meningeal arteries (p < 0.01). Indeed, a positive correlation was found between the vascular supply from pial-cortical arteries and the type of cleavage (p < 0.05). CONCLUSIONS: In this analysis the authors proved that there is a strong correlation between the amount of peritumoral edema, hyperintensity of the tumor on T2-weighted images, cortical penetration, vascular supply from pial-cortical arteries, and cleavage of the meningioma. Therefore, the consequent difficulty of microsurgical dissection can be predicted preoperatively by analyzing MR imaging and angiographic studies. PMID- 10470812 TI - Family history of subarachnoid hemorrhage and the incidence of asymptomatic, unruptured cerebral aneurysms. AB - OBJECT: Previously the authors reported a significant correlation between a family history of subarachnoid hemorrhage (SAH) and the discovery of an unruptured aneurysm in a group of healthy volunteers. This study corroborates and extends previous findings regarding the relationship between genetic and acquired factors in the formation of cerebral aneurysms. METHODS: The incidence of asymptomatic, unruptured cerebral aneurysms was studied among patients with a family history of SAH within the second degree of consanguinity. Forty-one unruptured cerebral aneurysms were found in 34 (13.9%) of 244 patients. This incidence was significantly higher than that found in a control group of healthy volunteers (6%). Furthermore, patients who had a family history of SAH combined with multiple systemic risk factors were found to have the highest incidence of unruptured aneurysms (32%; odds ratio 3.49, 95% confidence interval 1.37-8.9). CONCLUSIONS: These findings suggest that patients with a family history of SAH with or without the presence of more than one systemic risk factor are at significantly higher risk of harboring cerebral aneurysms. This high-risk group should be periodically screened and treated with appropriate surgical or other forms of therapy when necessary. PMID- 10470813 TI - Late angiographic follow-up review of surgically treated aneurysms. AB - OBJECT: This study was undertaken to evaluate the long-term angiographic outcome of surgically treated aneurysms, which is unknown. Specifically, the incidence of recurrent aneurysms, the fate of residual necks, and the de novo formation of aneurysms were evaluated. METHODS: One hundred two patients (80 females and 22 males; mean age 49 years; range 12-78 years) harboring a total of 167 aneurysms underwent late follow-up angiography; 160 aneurysms were surgically treated. Late angiographic follow-up review was obtained at a mean of 4.4 +/- 1.6 years postsurgery (range 2.6-9.7 years). Late follow-up angiography revealed two recurrent aneurysms (1.5%) of 135 clipped aneurysms without residua. Of 12 aneurysms with known residua, there were eight "dog-ear" residua, of which two (25%) enlarged. One hemorrhage was noted, yielding a hemorrhage risk of 1.9% per year. A second subgroup with broad-based residua revealed dramatic regrowth in three of four cases. Eight de novo aneurysms were found in six patients, for an annual risk of 1.8% per year. A history of multiple aneurysms was associated with de novo aneurysm formation (p = 0.049, chi-square analysis). CONCLUSIONS: This study confirms the long-term efficacy of aneurysm clip ligation. In addition, the authors found there is a small but significant risk of de novo aneurysm formation, particularly in patients with multiple aneurysms. Most residual aneurysm rests appear to remain stable, although a subset may enlarge or rupture. These findings support the rationale for late angiographic follow-up review in patients with aneurysms. PMID- 10470814 TI - Cognition following subarachnoid hemorrhage from anterior communicating artery aneurysm: relation to timing of surgery. AB - OBJECT: This study was conducted to define neuropsychological changes following operation for subarachnoid hemorrhage (SAH) caused by rupture of an anterior communicating artery (ACoA) aneurysm and to assess the influence of the timing of surgery to clip the aneurysm. METHODS: Cognitive outcome was evaluated using the Cambridge Neuropsychological Test Automated Battery in patients with an ACoA aneurysm that had caused an SAH. Adult patients younger than 70 years of age who had achieved a favorable neurological outcome (Glasgow Outcome Scale scores of 4 or 5) were studied 6 to 24 months postsurgery. Patients were divided into early (Days 0-3) and late surgery groups (after Day 3) according to the timing of surgery after the ictus. Neuropsychological analysis was performed by reviewers who were blinded to the timing of surgery. Forty-seven patients whose mean age was 51.5 years were tested. They were compared with age- and intelligence quotient (IQ)matched controls by using premorbid IQ as estimated on the National Adult Reading Test. Patients showed deficiencies in several tasks of verbal fluency, pattern recognition, and spatial working memory; this profile of deficits was similar to that seen in patients who underwent temporal lobe excisions. However, there was no significant difference in cognitive performance between the early and late surgery groups. CONCLUSIONS: After open surgery for ruptured ACoA aneurysms, patients who have achieved a favorable neurological outcome still exhibit significant cognitive deficits, primarily in tests sensitive to temporal lobe dysfunction. However, early surgery does not carry a higher risk of neuropsychological disability. PMID- 10470815 TI - Results of early surgical evacuation of packed intraventricular hemorrhage from aneurysm rupture in patients with poor-grade subarachnoid hemorrhage. AB - OBJECT: The goal of this study was to evaluate the results of early surgical evacuation of "packed" intraventricular hemorrhage (IVH) in patients with poor grade subarachnoid hemorrhage (SAH). METHODS: The authors performed surgery within 24 hours after onset of SAH, identified on neuroimaging as a cast distending the ventricular system, in 74 patients with poor-grade SAH (World Federation of Neurosurgical Societies Grades IV and V) without intracerebral hemorrhage. Eighteen of these patients had packed IVH; in these patients the intraventricular clots were extensively evacuated via frontal corticotomy performed under microscopic view. CONCLUSIONS: Overall, 42% of the 74 patients undergoing craniotomy in the acute stage had favorable outcomes, whereas 30% died. Using multivariate analysis, variables significantly associated with favorable outcome in patients with poor-grade SAH included absence of a packed intraventricular clot on computerized tomography scanning; absence of a history of cardiac disease; and a Glasgow Coma Scale score of 11 or 12. None of the 18 patients who had packed IVH had favorable outcomes and seven of these died. In six recently treated patients with packed IVH, which was examined using fluid attenuated inversion recovery imaging, extensive periventricular brain damage was found both immediately after surgery and during the chronic stage. Accordingly, the authors believe that irreversible periventricular brain damage is already complete immediately after packed IVH occurs. PMID- 10470816 TI - Percutaneous transluminal angioplasty for intracranial atherosclerotic lesions: evolution of technique and short-term results. AB - OBJECT: A retrospective analysis of a 9-year experience with balloon angioplasty for intracranial atherosclerotic stenosis was undertaken with the goals of illustrating development of a safe technique for treatment of intracranial atherosclerotic disease and reporting the immediate results in this series of patients. METHODS: Three distinct periods are defined, based on the technique used. In the early period, in which eight patients were treated, the angioplasty balloon size approximated the vessel size, but was always smaller. Angioplasty was moderately rapid and brief (15-30 seconds). Clinical improvement occurred in seven (87.5%) of eight patients, dissection without consequence occurred in four (50%) of eight, and residual stenosis greater than 50% was found in three (37.5%) of eight. No neurological complications occurred. In the middle period, in which 12 patients were treated, the balloon size approximated the vessel size, but oversizing by up to 0.25 mm was permitted. Angioplasty was extremely rapid and brief. Angiographically visible dissection occurred in nine (75%) of 12 patients, necessitating urokinase infusion in five (41.7%) of 12 and producing abrupt occlusion in one (8.3%) of 12, resulting in death. Occlusion secondary to the recrossing of the lesion occurred in one (8.3%) of 12, resulting in stroke. Good outcome was eventually achieved in 10 (83.3%) of 12. In the current period, in which 50 patients have been treated, the balloon is always undersized and inflation is extremely slow (several minutes). Dissection occurred in seven (14%) of 50 patients, necessitating fibrinolysis in two of 50 (4%, both uneventful) and producing no abrupt occlusion or stroke. Residual stenosis greater than 50% occurred in eight (16%) of 50, with no stenosis greater than 70%. Late restenosis occurred in four (9%) of 44 and successful repeated angioplasty was performed in all four. One guidewire vessel perforation occurred (2%), resulting in the patient's death. Good angiographic and short-term clinical outcome was achieved in the other 49 patients (98%). CONCLUSIONS: Extremely slow balloon inflation combined with balloon undersizing results in decreased intimal damage, decreased acute platelet/thrombus deposition, and decreased acute closure. This technique sometimes yields suboptimal angiographic results but achieves the clinical goal safely. Intracranial angioplasty can be safely performed using this technique and modern equipment. PMID- 10470817 TI - Three-dimensional computerized tomography angiography in patients with hyperacute intracerebral hemorrhage. AB - OBJECT: The authors confirm the usefulness of extravasation detected on three dimensional computerized tomography (3D-CT) angiography in the diagnosis of continued hemorrhage and establishment of its cause in patients with acute intracerebral hemorrhage (ICH). METHODS: Thirty-one patients with acute ICH in whom noncontrast and 3D-CT angiography had been performed within 12 hours of the onset of hemorrhage and in whom conventional cerebral angiographic studies were obtained during the chronic stage were prospectively studied. Noncontrast CT scanning was repeated within 24 hours of the onset of ICH to evaluate hematoma enlargement. Findings indicating extravasation on 3D-CT angiography, including any abnormal area of high density on helical CT scanning, were observed in five patients; three of these demonstrated hematoma enlargement on follow-up CT studies. Thus, specificity was 60% (three correct predictions among five positives) and sensitivity was 100% (19 correct predictions among 19 negatives). Evidence of extravasation on 3D-CT angiography indicates that there is persistent hemorrhage and correlates with enlargement of the hematoma. Regarding the cause of hemorrhage, five cerebral aneurysms were visualized in four patients, and two diagnoses of moyamoya disease and one of unilateral moyamoya phenomenon were made with the aid of 3D-CT angiography. Emergency surgery was performed without conventional angiography in one patient who had an aneurysm, and it was clipped successfully. CONCLUSIONS: Overall, 3D-CT angiography was found to be valuable in the diagnosis of the cause of hemorrhage and in the detection of persistent hemorrhage in patients with acute ICH. PMID- 10470818 TI - Proton radiation therapy for chordomas and chondrosarcomas of the skull base. AB - OBJECT: Local tumor control, patient survival, and treatment failure outcomes were analyzed to assess treatment efficacy in 58 patients in whom fractionated proton radiation therapy (RT) was administered for skull base chordomas and chondrosarcomas. METHODS: Between March 1992 and January 1998, a total of 58 patients who could be evaluated were treated for skull base tumors, 33 for chordoma and 25 for chondrosarcoma. Following various surgical procedures, residual tumor was detected in 91% of patients; 59% demonstrated brainstem involvement. Target dosages ranged from 64.8 and 79.2 (mean 70.7) Co Gy equivalent. The range of follow up was 7 to 75 months (mean 33 months). In 10 patients (17%) the treatment failed locally, resulting in local control rates of 92% (23 of 25 patients) for chondrosarcomas and 76% (25 of 33 patients) for chordomas. Tumor volume and brainstem involvement influenced control rates. All tumors with volumes of 25 ml or less remained locally controlled, compared with 56% of tumors larger than 25 ml (p = 0.02); 94% of patients without brainstem involvement did not experience recurrence; in patients with brainstem involvement (and dose reduction because of brainstem tolerance constraints) the authors achieved a tumor control rate of 53% (p = 0.04). Three patients died of their disease, and one died of intercurrent disease. Actuarial 5-year survival rates were 100% for patients with chondrosarcoma and 79% for patients with chordoma. Grade 3 and 4 late toxicities were observed in four patients (7%) and were symptomatic in three (5%). CONCLUSIONS: High-dose proton RT offers excellent chances of lasting tumor control and survival, with acceptable risks. In this series all small- and medium-sized tumors with no demonstrable brainstem involvement have been controlled; all such patients are alive. Surgical debulking enhanced delivery of full tumoricidal doses, but even patients with large tumors and disease abutting crucial normal structures benefited. PMID- 10470819 TI - Expression of the JAK and STAT superfamilies in human meningiomas. AB - OBJECT: The goal of this study was to investigate whether the janus kinase/signal transducer and activator of transcription (JAK/STAT) signal transduction pathway is present and active in meningiomas. The results of these investigations are important for all meningioma therapies that, similar to interferon-alpha-2B (IFNalpha-2B), depend on activation of this pathway for their effect. The authors were interested in evaluating the importance, if any, of the JAK/STAT pathway in the biology and therapy for these tumors. METHODS: Total proteins were extracted from 17 meningioma samples and the levels of JAKs and STATs were determined by using Western blot analysis. Levels of these proteins in meningiomas were compared with those found in normal dura. The JAKs and STATs (with the exception of Jak3 and Tyk2) were present both in the dura and in the meningiomas studied. In tumors JAK and STAT levels were always significantly higher than those found in normal dura. Differences in relative levels were found when meningiomas were subdivided according to the current neuropathological criteria and the highest levels were found in transitional meningiomas. The authors also investigated, using tyrosine-phosphorylated Statl and Stat3 antibodies, whether STATs were activated in meningiomas and normal dura in vivo. Their results indicate that both Statl and Stat3 are phosphorylated in vivo in meningiomas and in the dura. Furthermore, in vitro experiments in which two independent short-term cultures obtained from freshly dissected meningioma samples were used indicated that Statl and Stat3 are phosphorylated in response to treatment with IFNalpha-2B. Exposure of meningioma cells to IFNalpha-2B leads to nuclear translocation of tyrosine phosphorylated Statl and Stat3, as demonstrated by immunocytochemical analysis. CONCLUSIONS: The results of this study indicate that the JAK and STAT families of proteins are important effectors in brain tumors and support the idea that the effects of IFNalpha in vivo are direct and not mediated by the immune system. This suggests a role for modulation of STAT transcription factors in inhibiting meningioma cell proliferation. PMID- 10470820 TI - Intraventricular infusion of nerve growth factor as the cause of sympathetic fiber sprouting in sensory ganglia. AB - OBJECT: The results of previous clinical trials have indicated that intraventricular infusion of nerve growth factor (NGF) in patients with Alzheimer's disease is frustrated by the appearance of weight loss and diffuse back pain. The present study tested whether NGF induces sympathetic sprouting in sensory ganglia. Such sprouting has been implicated in previous studies as a possible mechanism of sympathetically maintained pain in neuropathic animals. METHODS: Nineteen Long-Evans rats underwent intraventricular infusion of either artificial cerebrospinal fluid (ACSF; seven animals) or NGF (12 animals). After 14 days of infusion, the sensory ganglia of the trigeminal nerve and the C-2, C 8, T-1, L-4, and L-5 dorsal roots were examined for sympathetic sprouting by using tyrosine hydroxylase immunohistochemical analysis. CONCLUSIONS: In the animals receiving NGF, 52 of 144 ganglia showed sympathetic fiber sprouting. In the control animals receiving ACSF, only two of 72 ganglia showed minor sympathetic fiber sprouting. A preferential sprouting of sympathetic fibers was demonstrated at lower lumbar ganglia compared with the cervical and thoracic ganglia. The data presented here demonstrate that in the rat intraventricular NGF infusion caused sympathetic sprouting in dorsal root ganglia (p < 0.01). These findings may have importance both for the treatment of Alzheimer's disease and the understanding of neuropathic pain. PMID- 10470821 TI - Use of cytological preparations for the intraoperative diagnosis of stereotactically obtained brain biopsies: a 19-year experience and survey of neuropathologists. AB - OBJECT: The goals of this study were to analyze the accuracy of cytological techniques, consisting of touch and smear preparations, for the intraoperative diagnosis of stereotactically obtained brain biopsy samples, and to determine the prevalence of the use of these methods among neuropathologists. METHODS: A survey regarding preferred methods for intraoperative diagnosis of stereotactically obtained brain biopsy samples was completed by 92 (62%) of 148 neuropathologists. Twenty-three percent of respondents chose frozen-section examination alone; 13% chose one or more cytological methods alone; and the remainder (64%) chose a combination of frozen-section examination and cytology. At the University of Pittsburgh, the neuropathology records for all stereotactic brain biopsies performed from May 1979 through May 1998 were retrospectively reviewed. Of the 946 stereotactic brain biopsies, 316 cases were excluded because the intraoperative neuropathological consultation was not recorded. Thirty-five cases were excluded because frozen-section examinations were performed. Therefore, a total of 595 cases were suitable for analysis. Intraoperative cytological investigation correlated with the final diagnosis in 90% of cases (52% complete correlation and 38% partial correlation). In 11% of cases there was no correlation between the intraoperative and final diagnoses. Intraoperative diagnoses were most accurate in cases of abscess, germinoma, lymphoma, metastasis, and malignant glioma. Overall, 91% of biopsy specimens were diagnostic when examined using the paraffin-embedded section technique. The sensitivity of cytological preparations in detecting a diagnostic specimen was 96% and the specificity in detecting a nondiagnostic specimen was 75%. CONCLUSIONS: Intraoperative cytological preparations correlated with the final diagnoses in 90% of stereotactic biopsies and had a 96% sensitivity in detecting diagnostic specimens. The highest rate of correlation was noted in cases of abscess, germinoma, lymphoma, metastasis, and malignant glial tumor. PMID- 10470822 TI - Oxyhemoglobin-induced apoptosis in cultured endothelial cells. AB - OBJECT: Oxyhemoglobin (OxyHb) is one of the most important spasmogens for cerebral vasospasm that follows aneurysmal subarachnoid hemorrhage. The cytotoxic effect of OxyHb has been documented in endothelial and smooth-muscle cells; however, the pattern of cell death--necrosis or apoptosis--as the final stage of cell damage has not been demonstrated. This study was undertaken to determine if OxyHb induces apoptotic changes in cultured bovine aortic endothelial cells. METHODS: Confluent bovine aortic endothelial cells were treated with OxyHb in a concentration- and time-dependent manner. Cell density was assayed by counting the number of cells that attached to culture dishes after exposure to OxyHb. To identify apoptotic changes, the investigators used three specific methods: DNA fragmentation (electrophoreses), the apoptotic body (transmission electron microscopy), and cleavage of poly (adenosine diphosphate ribose) polymerase (PARP [Western blotting]). CONCLUSIONS: Oxyhemoglobin decreased cell density in a concentration- and time-dependent manner. Analysis of DNA showed a pattern of internucleosomal cleavage characteristic of apoptosis (DNA ladder). Transmission electron microscopy demonstrated condensation of nuclei and apoptotic bodies in OxyHb-treated endothelial cells. Western blotting with the PARP antibody revealed that the 116-kD PARP was cleaved to the 85-kD apoptosis-related fragment. These results for the first time demonstrated that the OxyHb induces apoptosis in cultured endothelial cells. PMID- 10470823 TI - Topographic anatomy of the cochlear nuclear region at the floor of the fourth ventricle in humans. AB - OBJECT: The development of appropriate methods to stimulate the dorsal and ventral cochlear nucleus by means of an auditory brainstem implant in patients with acquired bilateral anacusis requires a detailed topoanatomical knowledge both of the location and extension of the nuclear surface in the fourth ventricle and lateral recess and of its variability. The goal of this study was to provide that information. Anatomically, it is possible to use a midline surgical approach to the fourth ventricle rather than the translabyrinthine and suboccipital routes of access used hitherto. This is especially useful if severe scarring, which occurs as a result of tumor removal in the cerebellopontine angle, make the orientation and placement of an auditory brainstem implant via a lateral surgical approach difficult. There have been only a few studies, involving single cases and small series of patients, in which the focus was the exact extension of the cochlear nuclei, whose microsurgically relevant position in relation to the surface structures is not known in detail. METHODS: Landmarks that are important for the placement of an auditory brainstem implant through the fourth ventricle were examined and measured in a large series of 28 formalin-fixed human brainstems. In all cases, these examinations were supplemented by addition of a histological section series. For the first time values of unfixed fresh brainstem tissue were determined. Anatomical features are discussed with regard to their possible neurosurgical relevance, taking into account inter- and intraindividual variability. CONCLUSIONS: The midline approach would provide an opportunity to stimulate the whole area of the dorsal as well as the ventral cochlear nucleus with an auditory brainstem implant. PMID- 10470824 TI - Immunohistochemical staining of DNA topoisomerase IIalpha in human gliomas. AB - OBJECT: The enzyme DNA topoisomerase IIalpha (Topo IIalpha) was tested as a measure of cell proliferation in gliomas. METHODS: Immunostaining for the Topo IIalpha and for the Ki-67 antigen (MIB-1 antibody) was performed in paraffin embedded tissue sections obtained from 25 resected human gliomas. Additionally, cultured human glioma cells were subjected to simultaneous flow cytometry to determine Topo IIalpha and DNA content. Using flow cytometric analysis, the authors found that the Topo IIalpha antibody labeled cells in the S, G2, and M phases of the cell cycle and also those in some parts of the G0 and G1 phases. In histological sections, Topo IIalpha showed more distinct staining than MIB-1, particularly in older archival cases. The proliferative indices (PIs) based on cells staining for MIB-1 and Topo IIalpha correlated highly with one another (r = 0.96). The Topo IIalpha PI immunopositivity was seen in 4.07% of cells in the low grade astrocytoma group, 11.97% in the anaplastic astrocytoma group, and 13.84% in the glioblastoma multiforme group, representing significant differences between low-grade astrocytoma and both anaplastic astrocytoma and glioblastoma. A Topo IIalpha PI less than 5% predicted longer patient survival (p = 0.003). CONCLUSIONS: Immunostaining for Topo IIalpha represents a useful alternative to MIB-1 as a proliferative index in human gliomas. PMID- 10470825 TI - Overexpression of bax in human glioma cell lines. AB - OBJECT: Cells that lose their ability to undergo apoptosis may promote the development of neoplasms and result in resistance to clinical treatment with DNA damaging modalities such as radio- and chemotherapy. Four established human glioma cell lines that are resistant to apoptosis were transfected with the proapoptotic gene bax and assessed for their sensitivity to a proapoptotic stimulus. METHODS: Two cell lines had a wild-type p53 genotype (U87 and D247MG) and two had mutant p53 genotypes (U138 and U373). Constitutive overexpression of murine bax was achieved in U138 and U373 only, which resulted in an increased sensitivity of these lines to the apoptosis-inducing effect of cytosine arabinoside (ara-C). Multiple attempts to produce constitutive overexpression of bax in U87 and D247MG cells resulted in spontaneous, near-complete cell loss. Vector-only control transfections were successful in all four cell lines. Inducible overexpression of bax was achieved in the U87 cells and elevated levels of BAX were observed as early as 6 hours after gene induction. This overexpression of BAX resulted in the spontaneous induction of apoptosis in these cells. CONCLUSIONS: Overexpression of BAX in four human glioma cell lines resulted in increased sensitivity to apoptosis. In the two lines that had a wild type p53 genotype, overexpression of BAX produced spontaneous apoptosis. In contrast, the lines that had mutant, nonfunctional P53 did not undergo spontaneous apoptosis, but they were rendered more sensitive to the apoptosis inducing effect of ara-C. Modulation of BAX expression may be a useful therapeutic modality for gliomas, regardless of p53 genotype. PMID- 10470826 TI - Gangliocytoma masquerading as a prolactinoma. Case report. AB - The authors describe the case of a 36-year-old man who presented with bitemporal hemianopsia and a serum prolactin concentration of 1440 ng/ml. Magnetic resonance imaging of the pituitary revealed a presumed macroadenoma with suprasellar and temporal lobe extension. Although the patient's prolactin level was lowered to 55 ng/ml by bromocriptine therapy, no tumor shrinkage occurred. Fourteen months later, progression of visual field defects necessitated transsphenoidal resection, which was incomplete. Immunocytochemical analysis of the biopsy tissue was positive for prolactin and, in view of the clinical picture, more detailed analysis was not performed. External-beam radiotherapy was given 2 years later because of enlargement of residual tumor. Subsequently, despite a fall in the serum prolactin concentration to less than 20 ng/ml in response to the course of bromocriptine, the mass displayed further extension into the temporal lobe. Nine years after the patient's initial presentation, he underwent transfrontal craniotomy for sudden deterioration in visual acuity caused by hemorrhage into the mass. No adenohypophyseal tissue was identified in the resected tissue. The mass was composed of dysplastic neurons that were strongly immunoreactive for synaptophysin and neurofilament (indicating neural differentiation) and prolactin. Review of the original biopsy specimen indicated that the prolactin positive cells had striking neuronal morphological characteristics. The final diagnosis in this case is prolactin-secreting gangliocytoma. Although exceedingly rare, this disease must be added to the differential diagnosis in cases of "prolactinoma" when bromocriptine therapy is followed by a marked decline in serum prolactin that is not accompanied by significant tumor shrinkage. Furthermore, in such instances, consideration should be given to "obtaining a biopsy sample prior to electing for radiotherapy. PMID- 10470827 TI - Meningioma recurrence at a different site masquerading as a subdural hematoma. Case report. AB - It is unusual for a meningioma to recur at a distant site. It is more unusual for a meningioma to present as a subdural hematoma. The authors report a unique case in which both of these events occurred in the same patient during a 4-year period. The authors discuss the pathogenesis of these rare events with regard to meningiomas. PMID- 10470828 TI - Infectious meningitis mimicking recurrent medulloblastoma on magnetic resonance imaging. Case report. AB - This report and the accompanying review of the literature address the challenges, when using surveillance magnetic resonance (MR) imaging, of establishing the origin of newly detected central nervous system lesions. Routine surveillance MR imaging in a 16-year-old boy, whose medulloblastoma had been successfully treated, demonstrated asymptomatic nodular leptomeningeal enhancement of the brain and spinal cord, which was consistent with recurrent disease. Examination of the cerebrospinal fluid, however, led to the diagnosis of bacterial meningitis. Two weeks after completion of antibiotic therapy, the original MR imaging findings were seen to have resolved. This case illustrates the importance of considering clinical and laboratory data, including results from a complete examination of the cerebrospinal fluid, when interpreting the origin of new lesions revealed by MR imaging. PMID- 10470829 TI - Suboccipital meningocele presenting as a huge retropharyngeal mass in a patient with neurofibromatosis Type 1. Case report. AB - The authors report an extremely rare case of neurofibromatosis Type 1 (NF1) with a suboccipital meningocele presenting as a huge retropharyngeal mass. A 73-year old woman with typical cutaneous manifestations of NF1 presented with nasal obstruction and dysphagia due to a retropharyngeal mass. Magnetic resonance imaging revealed a huge mass lesion extending from the right occipital bone defect to the retropharynx through the right paravertebral region. Computerized tomography scanning after intrathecal administration of contrast material confirmed that the mass was a meningocele protruding through a right occipital bone defect. The authors attempted to ligate this meningocele, most of which was excised via a suboccipital approach, but a second transcervical operation was required. Finally, the meningocele resolved and the patient was discharged without symptoms. PMID- 10470830 TI - Central neurocytoma of the fourth ventricle. Case report. AB - The authors report on a 17-year-old boy who suffered from slowly progressive and long-standing symptoms of ataxia, neck pain, and headache. Computerized tomography (CT) and magnetic resonance (MR) imaging revealed a tumor arising from the floor of the fourth ventricle that resulted in a moderate hydrocephalus. A partial resection was performed. Histological and immunohistological findings led to the diagnosis of an atypical central neurocytoma of the fourth ventricle. The imaging features on CT scanning, MR imaging, and proton MR spectroscopy studies, the clinical picture, and the prognosis of this very unusual tumor are discussed. Three cases of neurocytomas in the posterior fossa have been described to date; however, in all three cases some atypical aspects were present. In the present case, with the exception of the very unusual location, both imaging findings and clinical history perfectly met the definition of this rare tumor. PMID- 10470832 TI - Temporal glioblastoma causing trigeminal neuralgia. Case illustration. PMID- 10470831 TI - Endovascular occlusion of a carotid pseudoaneurysm complicating deep neck space infection in a child. Case report. AB - Pseudoaneurysm formation of the cervical internal carotid artery (ICA) is a rare, potentially lethal complication of deep neck space infection. This entity typically occurs following otolaryngological or upper respiratory tract infection. The pseudoaneurysm is heralded by a pulsatile neck mass, Homer's syndrome, lower cranial neuropathies, and/or hemorrhage that may be massive. The recommended treatment includes prompt arterial ligation. The authors present a case of pseudoaneurysm of the cervical ICA complicating a deep neck space infection. A parapharyngeal Staphylococcus aureus abscess developed in a previously healthy 6-year-old girl after she experienced pharyngitis. The abscess was drained via an intraoral approach. On postoperative Day 3, the patient developed a pulsatile neck mass, lethargy, ipsilateral Horner's syndrome, and hemoptysis, which resulted in hemorrhagic shock. Treatment included emergency endovascular occlusion of the cervical ICA and postembolization antibiotic treatment for 6 weeks. The patient has made an uneventful recovery as of her 18 month follow-up evaluation. Conclusions drawn.from this experience and a review of the literature include the following: 1) mycotic pseudoaneurysms of the carotid arteries have a typical clinical presentation that should enable timely recognition; 2) these lesions occur more commonly in children than in adults; 3) angiography with a view to performing endovascular occlusion should be undertaken promptly; and 4) endovascular occlusion of the pseudoaneurysm is a viable treatment option. PMID- 10470833 TI - Hemorrhagic colloid cyst in a patient with leukemia. Case illustration. PMID- 10470834 TI - Beating aneurysm sign: angiographic evidence of ruptured aneurysm tamponade by intracranial hemorrhage. Case illustration. PMID- 10470835 TI - Middle meningeal artery aneurysm. Case illustration. PMID- 10470836 TI - Nimodipine in subarachnoid hemorrhage. PMID- 10470837 TI - Intratumoral ethanol injection. PMID- 10470838 TI - Brainstem lesions after head injury. PMID- 10470839 TI - Brainstem lesions after head injury. PMID- 10470840 TI - Gemcitabine--a new agent in the treatment of non-small cell lung cancer. A commentary. PMID- 10470841 TI - Evaluation of a clinic-based screening program for lung cancer with a case control design in Kanagawa, Japan. AB - A case-control study was performed in Kanagawa Prefecture, Japan, which introduced lung cancer screening, by family physicians, to determine if this program was effective. The subjects were persons who died from lung cancer, restricted to those between 40 and 74 years old at death, and National Health Insurance (NHI) holders. A total of 193 lung cancer deaths meeting the criteria were selected for the case group. Three controls for each case were selected at random from living NHI holders matched by residence, gender and year of birth (+1 year). Smoking adjusted odds ratios were calculated using a multiple logistic regression model. The odds ratios of dying from lung cancer for screening participants within 12 months compared with non-participants was 0.535 which was statistically significant (95% CI, 0.337-0.850). The odds ratio in the 12-24 month period before diagnosis was 0.638 (0.302-0.967), which was also significant. The results demonstrated that screening for lung cancer by family physicians can reduce the lung cancer mortality, and that a clinic-based screening program by family physicians is effective. PMID- 10470842 TI - Alteration of the PTEN/MMAC1 gene locus in primary lung cancer with distant metastasis. AB - The PTEN/MMAC1 gene located at 10q23, has been proposed to be a tumor suppressor gene. To determine the involvement of alteration of the PTEN/MMAC1 gene in carcinogenesis and the progression of primary lung cancers, we analyzed tumor samples of primary and distant metastatic sites and normal lung tissue samples of 30 patients with advanced lung cancer with distant metastasis. The tissues were analyzed for allelic deletion and mutational inactivation of PTEN/MMAC1 by loss of heterozygosity (LOH) analysis, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), and direct sequence analysis. LOH of the PTEN/MMAC1 locus was common in each histologic type of primary lung cancer. In this study, the overall allelic deletion rate was 33.3% (7/21). Allelic loss at the primary site and that at the metastatic site of each patient, were identical; in most cases, it seemed that the allelic loss had occurred before metastasis. Sequence analysis of the PTEN/MMAC1 gene revealed a G to C substitution located 8 bp upstream of the coding region of exon 1 and which seems to be a polymorphism, in 4 of the 30 cases. Somatic mutations of the PTEN/MMAC1 gene were not identified in any of the tumors at the primary and metastatic sites. These data indicate that point mutations in the PTEN/MMAC1 gene are probably not an important factor in tumorigenesis and the progression of a major subset of lung cancers. Due to frequent allelic loss at the PTEN/MMAC1 locus occurring at a stage earlier than the metastatic process, alternative mechanisms in which the remaining allele is inactivated such as methylation or homozygous deletion of a small region of the gene that can not be detected by the usual analysis, or alteration of other important tumor suppressor genes lying close to the PTEN/MMAC1 gene on 10q23, may be involved in the tumorigenesis of lung cancers of all histologic subtypes. PMID- 10470843 TI - Human monoclonal antibody 28K29 highly reactive with lung adenocarcinomas of all grades of differentiation and with large cell carcinomas. AB - Human monoclonal antibody (mAb) 28K29 was previously established by fusing regional lymphocytes from a lung adenocarcinoma patient with human-mouse heteromyeloma. This mAb was shown to have an antigen localized in the membrane and cytoplasm of lung cancer cells. This mAb was investigated for reaction with tissue sections from formalin-fixed paraffin-embedded blocks from 100 patients with lung cancer. The mAb 28K29 reacted with 83% (5/6) of the well differentiated, 79% (22/28) of the moderately-differentiated, and 67% (4/6) of the poorly-differentiated lung adenocarcinoma sections. The mAb also reacted with 35% (14/40) of the squamous cell carcinoma sections, 70% (7/10) of the large cell carcinoma sections, and 20% (2/10) of the small cell carcinoma sections. In Western blot analyses, a broad band at a molecular weight of approximately 600 kDa was detected in extracts from the lung cancer tissues positive for immunohistostaining with the mAb 28K29. The results of the study suggest that the expression of the 28K29 antigen is independent of the histological differentiation grade in lung adenocarcinoma and that this antigen might be a useful marker for detection of both large cell carcinoma and adenocarcinoma of the lung as well as for investigation of the putative transition of large cell carcinoma to adenocarcinoma proposed by Yesner. PMID- 10470844 TI - Standard combination versus alternating chemotherapy in small cell lung cancer: a randomised clinical trial including 394 patients. 'Petites Cellules' Group. AB - PURPOSE: to compare standard and alternating administration of chemotherapy combinations in small cell lung cancer (SCLC) patients. MATERIAL AND METHODS: in a multicenter clinical trial, 394 previously untreated SCLC patients were randomised to receive, every 4 weeks, eight courses of either a standard regimen with CCNU, cyclophosphamide, adriamycin (CCA) and VP16 or an alternating regimen (CCA regimen alternating with cisplatin-vindesine-VP16). RESULTS: overall response rate was higher in the standard group (78%) than in the alternating group (64%) (P = 0.0001). Complete response rate was also higher in the standard group (32%) than in the alternating group (18%) (P = 0.004). The median survival in the overall SCLC population was 306 days in the standard group and 272 days in the alternating group (P = 0.08). In limited SCLC patients, median survival was higher in the standard group (421 days) than in the alternating group (328 days) (P = 0.01). Grade III/IV haematological toxicity was lower in patients in the alternating group (25 versus 47%) (P < 0.001). CONCLUSION: the standard regimen was better than the alternating regimen for patients with limited forms of SCLC. The alternating regimen, associated with better haematological safety and ensuring a fairly similar survival, may be considered in patients with extensive SCLC. Pleiomorphic resistance mechanisms to chemotherapy make it difficult to define a non-cross-resistant chemotherapy regimen. PMID- 10470845 TI - Clinical investigation of atypical adenomatous hyperplasia of the lung. AB - The clinicopathologic characteristics of atypical adenomatous hyperplasia (AAH) remain unclear. A total of 137 patients underwent resection for adenocarcinoma of the lung at our institution. Examination of resected lung tissue showed that in addition to adenocarcinoma AAH was present in 26 cases and was not present in 111 cases. All nonsmokers with AAH (n = 13) had earlier-stage disease (stage IA, IB, IIA, and IIB) and no history of respiratory disease. Among patients with stage IA disease, the relapse-free and overall survival curves for those with AAH (n = 14) tended to be better than for those without AAH (n = 40), but the difference was not statistically significant (P = 0.056 and 0.087, respectively). Concurrent presence of AAH may be a favorable prognostic indicator in patients with stage IA adenocarcinoma. PMID- 10470846 TI - Method revealing latent periodicity of the nucleotide sequences modified for a case of small samples. AB - An earlier reported method for revealing latent periodicity of the nucleotide sequences has been considerably modified in a case of small samples, by applying a Monte Carlo method. This improved method has been used to search for the latent periodicity of some nucleotide sequences of the EMBL data bank. The existence of the nucleotide sequences' latent periodicity has been shown for some genes. The results obtained have implied that periodicity of gene structure is projected onto the periodicity of primary amino acid sequences and, further, onto spatial protein conformation. Even though the periodic structure of gene sequences has been eroded, it is still retained in primary and/or spatial structures of corresponding proteins. Furthermore, in a few cases the study of genes' periodicity has suggested their possible evolutionary origin by multifold duplications of some gene's fragments. PMID- 10470847 TI - Mouse A9 cells containing single human chromosomes for analysis of genomic imprinting. AB - To develop an systematic in vitro approach for the study of genomic imprinting, we generated a new library of human/mouse A9 monochromosomal hybrids. We used whole cell fusion and microcell-mediated chromosome transfer to generate A9 hybrids containing a single, intact, bsr-tagged human chromosome derived from primary fibroblasts. A9 hybrids were identified that contained either human chromosome 1, 2, 4, 5, 7, 8, 10, 11, 15, 18, 20, or X. The parental origin of these chromosomes was determined by polymorphic analysis using microsatellite markers, and matched hybrids containing maternal and paternal chromosomes were identified for chromosomes 5, 10, 11 and 15. The imprinted gene KVLQT1 on human chromosome 11p15.5 was expressed exclusively from the maternal chromosome in A9 hybrids, and the parental-origin-specific expression patterns of several other imprinted genes were also maintained. This library of human monochromosomal hybrids is a valuable resource for the mapping and cloning of human genes and is a novel in vitro system for the screening of imprinted genes and for their functional analysis. PMID- 10470848 TI - Nucleotide sequence of psbQ gene for 16-kDa protein of oxygen-evolving complex from Arabidopsis thaliana and regulation of its expression. AB - The psbQ gene encoding a 16-kDa polypeptide of the oxygen-evolving complex of photosystem II has been isolated from Arabidopsis thaliana and characterized. The gene consists of a 28 nucleotide long leader sequence, two introns and three exons encoding a 223-amino-acid precursor polypeptide. The first 75 amino acids act as a transit peptide for the translocation of the polypeptide into the thylakoid lumen. Expression studies show that the gene is light-inducible and expresses only in green tissues with high steady-state mRNA levels in leaves. Using this gene as a probe, restriction fragment length polymorphism between two ecotypes, Columbia and Estland, has also been detected. PMID- 10470849 TI - A novel calmodulin-like protein gene in rice which has an unusual prolonged C terminal sequence carrying a putative prenylation site. AB - A rice cDNA encoding a novel calmodulin-like protein was identified. It has 38 additional amino acids at the C-terminus of a complete, typical calmodulin (CaM) sequence of 149 amino acids. The four C-terminal amino acid residues form a CAAL motif which could be a site for protein prenylation and may subsequently cause the protein to become membrane associated. RT-PCR analysis confirmed that such a combined protein gene truly exists in rice. Sequence analysis of its genomic counterpart showed that there is an intron located at junction of the normal CaM sequence and the 38 C-terminal amino acids. This introduces a potential stop codon for normal CaM if an alternative splicing mechanism is involved. Southern blot analysis of rice genomic DNA revealed that there is only one locus for this gene. The northern blot analysis showed that this gene is highly expressed in rice roots, shoots and flowers. The distribution of this protein demonstrates the functional importance of this novel CaM-like protein in rice. PMID- 10470850 TI - Structural analysis of Arabidopsis thaliana chromosome 5. IX. Sequence features of the regions of 1,011,550 bp covered by seventeen P1 and TAC clones. AB - In this series of projects sequencing the entire genome of Arabidopsis thaliana chromosome 5, non-redundant P1 and TAC clones have been sequenced according to the fine physical map, and as of May 7, 1999, the sequences of 16.2 Mb representing approximately 60% of chromosome 5 have been accumulated and released at our web site. In parallel, structural features of the sequenced regions have been analyzed by applying a variety of computer programs, and to date we have predicted a total of 2380 potential protein-coding genes in the 10,154,580 bp regions, which are covered by 142 P1 and TAC clones. In this paper, we newly analyzed the structural features of the 1,011,550 bp regions covered by additional 17 P1 and TAC clones, and predicted 298 protein-coding genes. The average density of the genes identified was 1 gene per 3394 bp. Introns were observed in 67% of the genes, and the average number per gene and the average length of the introns were 3.2 and 159 bp, respectively. The gene density became higher than the value estimated in the previously analyzed regions (1 gene per 4,267 bp), as the data in this paper were compiled based on a new standard of gene assignment including the computer-predicted hypothetical genes. The regions also contained 8 tRNA genes when searched by similarity to reported tRNA genes and the tRNA scan-SE program. The sequence data and information on the potential genes are available on the database KAOS (Kazusa Arabidopsis data Opening Site) at http://www.kazusa.or.jp/arabi/. PMID- 10470851 TI - Prediction of the coding sequences of unidentified human genes. XIV. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. AB - To extend our cDNA project for accumulating basic information on unidentified human genes, we newly determined the sequences of 100 cDNA clones from a set of size-fractionated human adult and fetal brain cDNA libraries, and predicted the coding sequences of the corresponding genes, named KIAA1019 to KIAA1118. The sequencing of these clones revealed that the average size of the inserts and corresponding open reading frames were 5.0 kb and 2.6 kb (880 amino acid residues), respectively. Database search of the predicted amino acid sequences classified 58 predicted gene products into the five functional categories, such as cell signaling/communication, cell structure/motility, nucleic acid management, protein management and cell division. It was also found that, for 34 gene products, homologues were detected in the databases, which were similar in sequence through almost the entire regions. The chromosomal locations of the genes were determined by using human-rodent hybrid panels unless their mapping data were already available in the public databases. The expression profiles of all the genes among 10 human tissues, 8 brain regions (amygdala, corpus callosum, cerebellum, caudate nucleus, hippocampus, substania nigra, subthalamic nucleus, and thalamus), spinal cord, fetal brain and fetal liver were also examined by reverse transcription-coupled polymerase chain reaction, products of which were quantified by enzyme-linked immunosorbent assay. PMID- 10470852 TI - Structure of rat gamma-tubulin and its binding to HP33. AB - Gamma-tubulin is localized at the microtubule organizing center and is thought to participate in the organizing of the microtubule network. In this study, we isolated a cDNA of rat gamma-tubulin. The rat gamma-tubulin cDNA encoded 451 amino acids, the same number as that of its counterpart in other vertebrates, and its structure was found to be highly conserved in vertebrates. In a previous work, we identified HP33 (hepatocarcinogenesis- and hepatocellular proliferation related 33-kDa protein) that was localized at the centrosome of hepatic cells and that exhibited MAP-like activity. In vitro GST pull-down assay using highly purified recombinant HP33 and bacterially expressed gamma-tubulin demonstrated that HP33 bound to gamma-tubulin directly. These results suggest that HP33 is localized at the centrosome via association with both the microtubule and its minus end-specific component, gamma-tubulin. PMID- 10470853 TI - The inhibitory effects of transforming growth factor beta1 on breast cancer cell proliferation are mediated through regulation of aberrant nuclear factor kappaB/Rel expression. AB - Nuclear factor (NF)-kappaB/Rel transcription factors normally exist in non-B cells, such as epithelial cells, in inactive forms sequestered in the cytoplasm with specific inhibitory proteins, termed IkappaBs. Recently, however, we discovered that breast cancer is typified by aberrant constitutive expression of NF-kappaB/Rel factors. Because these factors control genes that regulate cell proliferation, here we analyzed the potential role of NF-kappaB/Rel in the ability of transforming growth factor (TGF)-beta1 to inhibit the growth of breast cancer cells. The decreased growth of Hs578T and MCF7 breast cancer cell lines on TGF-beta1 treatment correlated with a drop in NF-kappaB/Rel binding. This decrease was due to the stabilization of the inhibitory protein IKB-alpha. Ectopic expression of c-Rel in Hs578T cells led to the maintenance of NF kappaB/Rel binding and resistance to TGF-beta1-mediated inhibition of proliferation. Similarly, expression of the p65 subunit ablated the inhibition of Hs578T cell growth mediated by TGF-beta1. Thus, the inhibition of the aberrantly activated, constitutive NF-kappaB/Rel plays an important role in the arrest of the proliferation of breast cancer cells, which suggests that NF-kappaB/Rel may be a useful target in the treatment of breast cancer. PMID- 10470854 TI - Inhibition of the Jun kinase pathway blocks DNA repair, enhances p53-mediated apoptosis and promotes gene amplification. AB - We have previously shown, by expression of a nonphosphorylatable dominant inhibitor mutant of c-Jun [cJun(S63A,S73A)], that activation of the NH2-terminal Jun kinase/stress-activated protein kinase by genotoxic damage is required for DNA repair. Here, we examine the consequences of inhibition of DNA repair on p53 induced apoptosis in T98G cells, which are devoid of endogenous wild-type p53. Relative to parental or wild-type c-Jun-expressing control cells, mutant Jun expressing T98G clones show similar growth rates and plating efficiencies. However, these cells are unable to repair DNA (PCR-stop assays) and exhibit up to an 80-fold increased methotrexate-induced colony formation due to amplification of the dihydrofolate reductase gene. Moreover, the mutant c-Jun clones exhibit increased apoptosis and elevated bax:bcl2 ratios on expression of wild-type p53. These results indicate that inhibition of DNA repair leads to accumulation of DNA damage in tumor cells with unstable genomes and this, in turn, enhances p53mediated apoptosis. PMID- 10470855 TI - The ras suppressor, RSU-1, enhances nerve growth factor-induced differentiation of PC12 cells and induces p21CIP expression. AB - The Rsu-1 Ras suppressor gene was isolated based on its ability to inhibit v-Ras transformation. Using Rsu-1 transfectants of the pheochromocytoma cell line PC12, we demonstrated previously that Rsu-1 expression inhibited Jun kinase activation but enhanced Erk2 activation in response to epidermal growth factor. In the present study, the Rsu-1 PC12 transfectants were used to investigate the role of Rsu-1 in nerve growth factor (NGF)- and v-Ki-ras-mediated neuronal differentiation. NGF-induced neurite extension was enhanced, not inhibited, by the expression of Rsu-1 in PC12 cells. The activation of Erk kinase activity in response to NGF was sustained longer in the Rsu-1 transfectants compared with the vector control cells. During NGF-mediated differentiation, an increase in the expression of specific mRNAs for the early response genes Fos, cJun, and NGF1a was detected in both the vector control and Rsu-1 transfectants. The expression of the differentiation-specific genes VGF8 and SCG10 was similar in Rsu-1 transfectants compared with the vector control cells. The induction of Rsu-1 expression in these cell lines did not inhibit v-Ki-ras-induced differentiation, as measured by neurite extension. These data suggest that although Rsu-1 blocked some Ras-dependent response(s), these responses were not required for differentiation. Moreover, the induction of Rsu-1 expression in the PC12 clones resulted in growth inhibition and p21(WAF/CIP) expression. Hence, Rsu-1 expression enhances NGF-induced differentiation while inhibiting the growth of cells. PMID- 10470856 TI - Ecotropic viral integration site-1 is activated during, and is sufficient for, neuroectodermal P19 cell differentiation. AB - Expression of the ecotropic viral integration site-1 (Evi1) proto-oncogene during murine embryonal development is observed by in situ hybridization in primary head folds and neural crest-derived cells associated with the peripheral nervous system and embryonic mesoderm. To elucidate whether expression of Evi1 is involved in early neuroectodermal or mesodermal differentiation, we used murine embryonal carcinoma P19 cells as a model for the study of early embryonic differentiation. After retinoic acid (RA) treatment with aggregation, expression of Evi1 was detected during neural differentiation in P19 cells. However, Evi1 was not expressed in P19 cells during mesodermal differentiation after DMSO treatment with aggregation. Enforced expression of Evi1 in P19 cells induced neuron-specific microtubule-associated protein-2 microtubule-associated protein-2 and TrkA expression in the absence of RA under monolayer culture. After incubation with RA with aggregation, the Evi1 clones expressed microtubule associated protein-2 continuously but did not express glial fibrillary acidic protein as an astrocyte marker protein until 12 days of culture. Thus, the overexpression of Evi1 leads to neural differentiation of P19 cells and blocks further differentiation into astrocytes by RA treatment, suggesting that Evi1 might be an important transcription factor for regulation of early neuroectodermal differentiation. PMID- 10470857 TI - Interferon-alpha inhibits proliferation in human T lymphocytes by abrogation of interleukin 2-induced changes in cell cycle-regulatory proteins. AB - IFN-alpha exerts prominent regulatory functions on the immune system. One such effect is the inhibition of proliferation of in vitro stimulated T lymphocytes. The exact physiological function of this activity is not known, but it has been implicated in the antiviral effects of IFN, its antitumor action in T-cell malignancies, and the regulation of the in vivo T-cell response. Here, we have investigated the mechanism underlying the IFN-alpha-mediated growth inhibition of normal human PHA- and IL-2-stimulated T lymphocytes by an analysis of how IFN alpha treatment influences known molecular events that normally accompany the transition from quiescence to proliferation in these cells. IFN-alpha treatment was found to profoundly block S-phase entry of stimulated T lymphocytes. This correlated with a strong inhibition of IL-2-induced changes in G1-regulatory proteins, including the prevented up-regulation of G1 cyclins and cyclin dependent kinases as well as an abrogation of mitogen-induced reduction of p27Kip1 levels. This latter effect was due to a maintained stability of the p27Kip1 protein in the IFN-alpha-treated cells. In line with these findings, phosphorylation of the pocket proteins was abrogated in IFN-alpha-treated cells. Furthermore, our data indicate that IFN-alpha has selective effects on the pathways that emerge from the IL-2 receptor because IFN-alpha treatment does not block IL-2-induced up-regulation of c-myc or Cdc25A. PMID- 10470858 TI - Influence of prolactin on the differentiation of mouse B-lymphoid precursors. AB - Development and activation of immune cells are submitted to hormonal influences, as illustrated by the roles of corticosteroids in thymus, pregnancy-related estrogens in B-cell development, or prolactin (PRL) on T-cell generation and function. We have analyzed the putative role of PRL in B lymphopoiesis and differentiation. We chose as an experimental model the interleukin (IL)-3 dependent BaF-3 pro-B cell line, which was transfected with the rat long form of the PRL receptor (PRL-R) and transferred from IL-3- to PRL-enriched media. When stimulated with PRL, the PRL-R transfectants underwent some changes characteristic of B-cell differentiation: (a) IL-2R alpha chain became positively controlled by PRL; (b) antiapoptotic Bcl-2 protein was induced by PRL in a dose dependent manner; and (c) transcription of the pre-B cell receptor encoding the lambda5 gene was strongly up-regulated. We attempted to evaluate the differentiation-promoting activity of PRL in more physiological conditions, and the presence of PRL-R in bone marrow B-cell precursors was revealed. Furthermore, PRL promoted significant expansions of defined B-lineage cell populations in short-term bone marrow cell cultures. These findings suggest that PRL, in collaboration with other cytokines and hormonal influences, modulates B-cell development. PMID- 10470859 TI - Mannose 6-phosphate/insulin-like growth factor II receptor mediates the growth inhibitory effects of retinoids. AB - Both retinoids and the mannose 6-phosphate/insulin-like growth factor-II receptor (M6P/IGF2R) have been shown to play an important role in controlling cell growth during embryonic development and oncogenesis. Our recent work (Kang et al., Proc. Natl. Acad. Sci. USA, 94: 13671-13676, 1997; Kang et al., Proc. Natl. Acad. Sci. USA, 95: 13687-13691, 1998) revealed a direct biochemical interaction between retinoic acid (RA) and the M6P/IGF2R, thereby leading us to hypothesize that the M6P/IGF2R may mediate a growth-inhibiting effect of RA. To test this hypothesis, cell growth and apoptosis in response to RA and various receptor-selective retinoids were examined in cells that lack or overexpress the M6P/IGF2R. RA and those retinoids capable of binding to the M6P/IGF2R induced a remarkable morphological change with characteristics of round shape and reduced spreading, apoptosis, and growth inhibition in stably transfected mouse P388D1 cells overexpressing the M6P/IGF2R but not in the M6P/IGF2R-deficient P388D1 cells. These effects of RA were neither blocked by a potent RA nuclear receptor (RAR) antagonist (AGN193109), nor mimicked by a selective RAR agonist (TTNPB), suggesting that the observed effects of RA are independent of RARs. Similar effects of the retinoids were observed in cultured neonatal rat cardiac myocytes that have high levels of the M6P/IGF2R. Furthermore, overexpression of the M6P/IGF2R in a RA-resistant cancer cell line (HL-60R) that lacked functional RARs gave the cells a susceptibility to RA-induced apoptosis. These data suggest that the M6P/ IGF2R may play an important role in mediating retinoid-induced apoptosis/growth-inhibition and provide insight into the similar biological effects of RA and the M6P/IGF2R on fetal development and carcinogenesis. PMID- 10470860 TI - Heart failure and angiotensin converting enzyme inhibition: problems and perspectives. AB - Heart failure has become the most widely studied syndrome in cardiology over the recent years. Despite the encouraging achievements by angiotensin converting enzyme (ACE) inhibitors, the mortality of patients with chronic heart failure remains high. There are several factors which can potentially be responsible for the fact that about 80% of patients with a failing heart defy protection by ACE inhibitors: different activation of tissue and systemic renin-angiotensin system (RAS) in a particular heart disease and the distinct ability of various ACE inhibitors to block cardiac ACE, alternative pathways for angiotensin II formation (chymase), genetic polymorphism of the RAS system and the complexity of neuroendocrine activation. Moreover, chronic heart failure can provoke disturbances in the reactivity of peripheral vessels and metabolism of striated muscles. These factors may then potentiate the vicious circle of heart failure. New therapeutic approaches, which could further reduce the mortality in patients with heart failure involve angiotensin II type 1 receptor antagonists, beta blockers, aldosterone antagonists and blockers of the endothelin receptor. A number of questions associated with functions of the RAS still remain open and their solution could be of substantial benefit for patients with a failing heart. PMID- 10470861 TI - Detrimental subtype-specific endothelin-1 (ET-1) signaling in myocardial cells: the ET(A) mediated proliferation and ET(B)receptor down-regulation. AB - Many physiological and pathological processes in the cardiac tissue have been shown to be associated with a release of endothelin (ET) peptides and with induction of specific ET-receptors and G-protein-coupled ion channels. However, the exact mechanism regulating ET-receptors in the myocardium is controversial. The response to ET-1, the most important member of the ET family, is rapidly attenuated by down-regulation of ET-receptors. The internalization of ET-1 bound to two subclasses of specific receptors (ET(A) and ET(B)) that are abundant in the myocardium has been hypothesized to activate and/or inhibit a variety of intracellular signal transducing systems. The [125I]ET-1, BQ-3020 and selective ET-antagonists were used to study the subtype-selective component of regulation of ET-1 receptors in myocardial membranes. We determined the characteristics of [125I]ET-1 binding and [3H]thymidine incorporation in whole cell saturation studies and measured Ca2+ channel induction and the total number of inactive Ca2+ channels in photoaffinity studies with [3H]azidopine. Here we demonstrate four important components of the complex ET-1 response in human, porcine and rat myocardium, leading to aberrant responses of cells. After ET-1 induction, adaptive subtype-ETB selective down-regulation predominated in human embryonic fibroblasts, in porcine membrane vesicles and in microsomal membranes of renal hypertensive rats, with preferential high affinity ET-1 binding to ETA receptors and with the resultant ETA mediated proliferative and mitogenic activation of human fibroblasts. The ET-1 induction was also accompanied by profound inactivation of Ca2+ channels in myocardial membranes. PMID- 10470862 TI - Control of breathing and brain activation in human subjects seen by functional magnetic resonance imaging. AB - Functional magnetic resonance imaging (fMRI) was used to demonstrate the brain activation during transition from unconscious to conscious breathing in seven healthy human subjects. In right-handed volunteers, the activated areas were found in both hemispheres. The medial part of the precentral gyrus (area 4) was constantly activated in the left hemisphere. Additional activated areas were demonstrated in the premotor cortex and in the posterior parietal cortex. The activated cortical sites exhibited analogous distribution in the right hemisphere. In two out of the seven subjects. activated sites were also observed in the cerebellar hemispheres, and in the lentiform and caudate nuclei. PMID- 10470863 TI - Creatine kinase reaction in skinned rat psoas muscle fibers and their myofibrils. AB - The aim of this study was to evaluate myofibrillar creatine kinase (EC 2.7.3.2) activity on the background of the effect of substrate channeling by myosin ATPase and to compare it with creatine kinase (CK) activity of whole skinned fibers. In order to assess CK activity, skinned fibers were prepared from the rat psoas major muscles defined by light microscopy. The activity in permeabilized fibers after treatment with saponin, Triton X-100 and Ca(2+)-free medium reached 2.80, 6.97 and 3.32 micromol ATP min(-1) mg(-1) protein, respectively, when a coupled enzyme assay system with external hexokinase and glucose-6-phosphate dehydrogenase was used. Transmission electron microscopy (TEM) revealed a possible interference among activities of sarcolemmal, sarcoplasmic, myofibrillar and mitochondrial CK from persisting structures. For evaluation of the myofibrillar CK itself, a pure myofibrillar fraction was prepared. Fraction purity was confirmed by TEM and by enzymatic assays for marker enzymes. Two procedures, i.e. the coupled enzyme assay and the evaluation of phosphocreatine (PCr) concentration before and after the CK reaction, were used for measurement of CK activity in this fraction. The procedures resulted in 3.2 nmol ATP min(-1) mg(-1) protein and 7.6 nmol PCr min(-1) mg(-1) protein, respectively. These alternative approaches revealed a discrepancy between the reacting portions of PCr by more than 50 %, which provides information about the size of the effect, generally described as substrate channeling. PMID- 10470864 TI - Granulocyte colony-stimulating factor and drugs elevating extracellular adenosine act additively to enhance the hemopoietic spleen colony formation in irradiated mice. AB - The effects of combined administration of two drugs elevating extracellular adenosine, namely dipyridamole (DP) and adenosine monophosphate (AMP), and granulocyte colony-stimulating factor (G-CSF) on hemopoietic stem cells in vivo were investigated. The experiments were performed on mice using the endogenous spleen colony formation in gamma-irradiated animals as an endpoint. The results have shown that DP and AMP act additively with G-CSF to enhance spleen colony formation and thus the erythroid repopulation of the spleen. These findings indicate that the signaling pathways of G-CSF and drugs elevating extracellular adenosine can interact at the level of primitive hemopoietic stem cells. The enhancement of hemopoiesis-stimulating effects of G-CSF by DP and AMP, which are low-priced and clinically available drugs, could improve the cost-effectiveness of the therapy with G-CSF. PMID- 10470865 TI - Chronic effects of somatotropin treatment in vivo and in vitro on lipogenic activity of goat adipose tissue in a glucose-free buffer during acute incubation. AB - Young castrated male goats (n = 8) were used to investigate the effect of long term treatment with recombinant methionyl bovine somatotropin in a sustained release vehicle (bST; 100 mg at seven-day intervals in a 147-day experiment) and chronic culture (24 h) of omental adipose tissue in the presence of various hormones on lipogenic responses to catecholamines during acute incubation (2 h) in a sodium acetate supplemented glucose-free buffer. The rate of fatty acid synthesis in freshly-prepared adipose explants was low and did not differ from those cultured in the absence of hormones for 24 h. Hormonal combination of insulin (17 nmol.l(-1)) plus cortisol (138 nmol.l(-1)) or insulin plus recombinant enterokinase linker bST (4.5 nmol.l(-1) increased lipogenesis (P<0.05). Further addition of bST or cortisol decreased lipogenesis significantly (P<0.05) in the controls but not significantly in bST-treated animals. Cultured explants from either control or bST-treated animals showed significant inhibition of lipogenesis by both norepinephrine (10 micromol.l(-1)) and isoprenaline (10 micromol.l(-1)). BST treatment in vivo did not increase the responsiveness of cultured explants to norepinephrine in vitro, however, the responsiveness to isoprenaline(inhibition of lipogenesis) was greater in bST-treated animals than in the controls. PMID- 10470866 TI - Changes of insulin binding in rat tissues after exposure to stress. AB - The effects of various stressors on insulin receptors in adipose, liver and skeletal muscle tissues were studied in rats exposed to acute or repeated stress. Adult male rats were exposed to immobilization (IMO) for 2.5 hours daily for 1, 7 and 42 days, or to hypokinesia (HK) for 1, 7 and 21 days. We determined the values of specific insulin binding (SIB) and insulin receptor binding capacity (IR) of plasma cell membranes from adipose, liver and muscle tissue (IMO groups), or insulin binding to isolated adipocytes and hepatocytes (HK groups). A significant decrease of SIB and IR was observed in rats exposed to acute stress (1x IMO) in muscle, adipose and liver tissues. However, in animals exposed to repeated stress (7x and 42x IMO), SIB and IR were diminished in the muscle tissue, whereas no significant changes were noted in the liver and adipose tissue. When tissue samples were collected 3-24 hours after exposure to IMO stress, no changes of SIB and IR were found in liver and adipose tissue, but insulin binding was lowered in skeletal muscles. In animals exposed to HK for one day, a decrease of SIB and IR was found in isolated adipocytes, but no changes in insulin binding were noted in the liver tissue. In rats exposed to HK for 7 and 21 days, values of IR were similar as in control group. Our results indicate a) the different changes of IR in the liver, fat and muscle tissues after exposure to stress situations, b) a long-term decrease of insulin binding in muscles of rats exposed to repeated IMO stress, and c) the return of reduced SIB and IR (induced by acute stress) to control values in the liver and adipose tissue after a short recovery period. PMID- 10470867 TI - The effect of branched chain amino acids on protein synthesis in two skeletal muscles of Japanese quail. AB - Two different isolated skeletal muscles of Japanese quail were used. The influence of branched chain amino acids on the fractional rate of protein synthesis (FSR) was evaluated using 14C-tyrosine. The addition of 0.5 mM valine, leucine or isoleucine to the incubation medium significantly decreased (P<0.05) the value of FSR in extensor metacarpalis radialis. In the ambiens muscle only the application of leucine increased the FSR significantly while valine and isoleucine were without any effect. PMID- 10470868 TI - Effects of histamine H1 antagonist dithiaden on acetic acid-induced colitis in rats. AB - To assess the possible involvement of mast cells and/or their mediators in inflammatory bowel diseases, the effect of the histamine H1 antagonist Dithiaden was studied on a model of acetic acid-induced colitis in rats. Dithiaden pretreatment by intracolonic administration was found to reduce the extent of acute inflammatory colonic injury. This was manifested by a decrease in the score of gross mucosal injury, by lowered colonic wet weight and by diminished myeloperoxidase activity reflecting reduced leukocyte infiltration. Vascular permeability and gamma-glutamyl transpeptidase activity, elevated by acetic acid exposure, were decreased after Dithiaden pretreatment. The results indicate that locally administered Dithiaden may protect the colonic mucosa against an acute inflammatory attack by interfering with the action of the major mast cell mediator histamine. PMID- 10470869 TI - The role of membrane fluidity changes and thiobarbituric acid-reactive substances production in the inhibition of cerebral cortex Na+/K+-ATPase activity. AB - Lipid peroxidation of rat cerebral cortex membranes was induced by Fe2+/ADP and ascorbate. The rate of Na+/K(+)-ATPase inhibition was correlated with the increase of thiobarbituric acid-reactive substances (TBARS) and conjugated dienes (CD) and with membrane fluidity changes. Our data showed that membrane fluidity changes (evaluated by fluorescence steady-state anisotropy measurements) can participate in Na+/K(+)-ATPase inhibition during the initial period of lipid peroxidation process, whereas during the following period the enzyme inhibition correlates only with TBARS and CD production. PMID- 10470870 TI - The effect of goldthioglucose on peroxidative processes in mice. AB - Experiments were conducted to study the effect of goldthioglucose (GTG) upon the processes associated with lipid peroxidation. The glucose-6-phosphate dehydrogenase activity (G6PD; E.C. 1. 1.1.49) in red blood cells (RBC) and the amount of malonaldehyde precursors (MDA) per gram of brain, liver and kidney were determined. Adult mice received i.p. injections for three consecutive days of either saline (controls) or GTG dissolved in saline, in a dose of 0.10 mg.g(-1) or 0. 15 mg.g(-1) b.w. In mice receiving higher dose of GTG the G6PD activity was significantly increased (349.38+/-17.46 mU.10(-9) RBC compared to 258.2+/-14.46 mU.10(-9) RBC in control animals). The content of MDA precursors rose significantly from 4.8+/-0.81 micromol.g(-1) of the liver in controls to 8.12+/ 1.41 micromol.g(-1) and 7.88+/-0.51 micromol.g(-1) and from 18.71+/-1.01 micromol.g(-1) of the kidneys in controls to 24.25+/-1.25 micromol.g(-1) and 24.88+/-1.7 micromol.g(-1) respectively. The GTG-induced higher levels of MDA precursors and increased G6PD activity in RBC corresponds to the rise in lipid peroxidation and its participation in producing the lesions after experimental and therapeutic use of gold-containing substances seems possible. PMID- 10470871 TI - Tumor necrosis factor alpha in various tissues of insulin-resistant obese Koletsky rats: relations to insulin receptor characteristics. AB - Tumor necrosis factor alpha (TNFalpha) was found to be significantly increased in skeletal muscles and retroperitoneal fat of obese insulin-resistant Koletsky rats as compared to control Wistar rats. This increase was accompanied by a depression of insulin receptor protein tyrosine kinase (PTK) activity. Neither the insulin binding capacity nor insulin receptor affinity were related to this TNFalpha increase in these tissues. In the liver, no significant changes of TNFalpha content and only a lowering of insulin-binding capacity were found. It is concluded that an increased TNFalpha content in muscles and fat (but not in the liver) contributes to insulin resistance by lowering insulin receptor protein tyrosine kinase activity, while other insulin receptor characteristics (insulin binding capacity and affinity of insulin receptors to the hormone) do not seem to be influenced by this factor. PMID- 10470872 TI - Kurt Amplatz: a radiology giant steps aside. PMID- 10470873 TI - A comprehensive approach using MR imaging to diagnose acute segmental mesenteric ischemia in a porcine model. AB - OBJECTIVE: Acute mesenteric ischemia is a lethal disease that lacks a noninvasive diagnostic test. We evaluated the abilities of contrast-enhanced MR angiography, MR oximetry, and real-time interactive MR imaging to diagnose segmental mesenteric ischemia in a porcine model. MATERIALS AND METHODS: Segmental mesenteric ischemia was created by subselective Gelfoam embolization of the mesenteric circulation in eight pigs. Conventional digital subtraction angiography (DSA), MR oximetry, and real-time interactive MR imaging of the small bowel were performed before and after embolization. Changes in the perfusion pattern seen on DSA established the regions of true ischemia. Postembolization DSA and MR angiography were compared with this gold standard. RESULTS: Both MR angiography and DSA had high sensitivity (91% and 100%, respectively) for detecting ischemic regions. The difference was not statistically significant (p > .2). MR angiography yielded lower specificity than DSA (80% and 90%, respectively; p < .01). After embolization, the oxygen saturation in the superior mesenteric vein (SMV) dropped significantly (p < .005). After embolization, the SMV also showed oxygen saturation significantly lower than that in the inferior vena cava (p < .005). In two of the animals, segmental hypomotility of the small bowel was observed. CONCLUSION: MR oximetry is capable of detecting oxygen desaturation caused by segmental ischemia. A loss of oxygen saturation in the SMV relative to that in the inferior vena cava provides a convenient marker of mesenteric ischemia. Contrast-enhanced MR angiography has sensitivity and specificity approaching those of DSA. Both MR techniques hold promise for the detection of acute mesenteric ischemia. PMID- 10470874 TI - Characteristics of women radiologists in the United States. AB - OBJECTIVE: Although women radiologists are increasingly prevalent, ways in which they differ from other women physicians are poorly described. We compared women radiologists with women physicians in general to see if and how they differed in personal and professional traits. MATERIALS AND METHODS: We analyzed data from the Women Physicians' Health Study, a nationally representative random sample (n = 4501 respondents) of personal and professional characteristics of women physicians in the United States. RESULTS: Radiologists (n = 163; 3.3% [weighted] of respondents) were more likely than were other women physicians to be conservative but were similar to other physicians in age, ethnicity, marital status, and personal health habits. Women radiologists were more likely to be board-certified and hospital-based, were not avid prevention counselors, and worked more hours than did other women physicians. Women radiologists had far higher incomes than other women physicians, but had less career satisfaction and less work control, and were more likely to feel overworked. Radiologists were also more likely than other physicians to report gender-based harassment while in medical school and postgraduate training, and more likely to report sexual harassment while in practice. CONCLUSION: Women radiologists are similar to other women physicians in some personal and professional characteristics but differ in some important and disturbing ways regarding professional experiences and satisfaction. PMID- 10470875 TI - Radiology 1998: Are today's residents ready for (tomorrow's) practice? PMID- 10470876 TI - Off-label use of interventional medical devices. PMID- 10470877 TI - Metallic stents: evaluation of MR imaging safety. AB - OBJECTIVE: The objective of our investigation was to evaluate safety during MR imaging (i.e., magnetic field interactions, heating, and artifacts) for metallic stents. MATERIALS AND METHODS: Different types of metallic stents were tested for magnetic field interactions, heating, and artifacts using a 1.5-T MR system. Magnetic field-related translational attraction and torque were assessed using previously described techniques. Heating was evaluated using an infrared thermometer to record temperatures immediately before and after performing MR imaging using a whole-body-averaged specific absorption rate of 1.3 W/kg. Artifacts were assessed by placing the stents inside a fluid-filled phantom and performing MR imaging using fast spoiled gradient-echo and T1-weighted spin-echo pulse sequences. RESULTS: For the 10 different stents evaluated, we found no magnetic field interactions. the highest temperature change was < or = +0.3 degrees C, and the artifacts involved signal voids that would not create diagnostic problems as long as the area of interest was not positioned exactly where a particular stent was located. CONCLUSION: The findings of the safety tests indicated that the 10 different metallic stents would be safe for patients undergoing MR imaging procedures using MR systems with static magnetic fields of 1.5 T or less. PMID- 10470878 TI - Helical CT colonography (virtual colonoscopy): the challenge that exists between advancing technology and generalizability. PMID- 10470879 TI - Usefulness of CT colonography in patients with incomplete colonoscopy. AB - OBJECTIVE: Our objective was to investigate the use of CT colonography in patients who have undergone incomplete colonoscopy. CONCLUSION: CT colonography is effective in evaluating portions of the colon not seen during colonoscopy and may have an adjunctive role. PMID- 10470880 TI - First International Symposium on Virtual Colonoscopy. PMID- 10470881 TI - The northern exposure sign: a newly described finding in sigmoid volvulus. AB - OBJECTIVE: Previous studies of sigmoid volvulus have focused on the migration and dilatation of the sigmoid with respect to both fixed and mobile landmarks in the abdomen. None has specifically referred to the relationship of this colonic segment to the more proximal large intestine. We analyzed findings on abdominal radiographs, all of which had been obtained with the patient supine, of sigmoid volvulus, with particular attention to the juxtaposition of the sigmoid colon with the transverse colon. MATERIALS AND METHODS: The abdominal radiographs of 30 patients with clinically confirmed sigmoid volvulus were obtained from the teaching files of four hospitals and were retrospectively reviewed. These radiographs were compared with abdominal radiographs of 28 individuals, each of whom had a dilated colon but not sigmoid volvulus. All radiographs had been obtained with the patient in the supine position. RESULTS: The transverse colon was identified in 26 of the 30 patients with sigmoid volvulus. In each of these 26 patients, the sigmoid colon was cephalad to the transverse colon. Of the patients in the control group, the transverse colon was identified in 24 of the 28 patients. In none of these control group patients did the sigmoid colon extend rostral to the transverse colon. Thus, this sign had a sensitivity of 86% and a specificity of 100%. CONCLUSION: A dilated sigmoid colon that ascends cephalad to the transverse colon is a newly described and accurate finding of sigmoid volvulus on abdominal radiographs obtained with the patient supine. PMID- 10470882 TI - The abdominal compartment syndrome: CT findings. AB - OBJECTIVE: The abdominal compartment syndrome is a potentially fatal condition resulting from pathologic elevation of intraabdominal pressure. We evaluated preoperative abdominal CT scans of four patients with proven abdominal compartment syndrome to identify signs of increased intraabdominal pressure. CONCLUSION: CT findings common to all four patients included tense infiltration of the retroperitoneum out of proportion to peritoneal disease, extrinsic compression of the inferior vena cava by retroperitoneal hemorrhage or exudate, and massive abdominal distention with an increased ratio of anteroposterior-to transverse abdominal diameter (positive round belly sign; ratio > .80; p < .001). Direct renal compression or displacement, bowel wall thickening with enhancement, and bilateral inguinal herniation were each present in two of the four patients. Radiologists should be aware of this life-threatening syndrome. In the appropriate clinical setting, CT findings of increased intraabdominal pressure should be swiftly communicated to other physicians involved in treating the patient because the abdominal compartment syndrome requires emergent surgical decompression. PMID- 10470883 TI - MR imaging of the small bowel using water as a contrast agent in a preliminary study with healthy volunteers. PMID- 10470884 TI - Dynamic contrast-enhanced MR imaging and dual-phase helical CT in the preoperative assessment of suspected pancreatic cancer: a comparative study with receiver operating characteristic analysis. AB - OBJECTIVE: The purpose of this study was to compare dynamic contrast-enhanced MR imaging and dual-phase helical CT in the preoperative assessment of patients with suspected pancreatic carcinoma. SUBJECTS AND METHODS: Thirty-three consecutive surgical candidates (20 men, 13 women; 39-81 years old) were included. MR imaging comprised fast spin-echo (TR/TE 4000/91), fat-suppressed T1-weighted spin-echo (500/15), and T1-weighted breath-hold gradient-echo fast low-angle shot (100/4; flip angle, 80 degrees) images before and after the administration of gadopentetate dimeglumine. Helical CT used 5-mm collimation with a pitch of 1:1.5 1.7; images were obtained 20 and 65-70 sec after injection of 150 ml of contrast material. Two pairs of interpreters who were unaware of the results of the other imaging method independently scored each examination for the presence of a lesion and for surgical resectability using a five-point scale. Results were correlated with surgery (n = 25) or consensus review (n = 8). Receiver operating characteristic methodology was used to analyze the results for resectability, and positive predictive values were calculated. RESULTS: Both MR imaging and helical CT revealed 29 of 31 lesions. In determining lesion resectability, the mean areas under the receiver operating characteristic curves were 0.96 and 0.81 (p = .01) and the positive predictive values were 86.5% and 76% (p = .02) for MR imaging and helical CT, respectively. CONCLUSION: MR imaging and helical CT performed equally well in lesion detection. MR imaging was significantly better in the assessment of resectability of pancreatic tumors. PMID- 10470886 TI - Contrast enhancement of small hepatocellular carcinoma: usefulness of three successive early image acquisitions during multiphase dynamic MR imaging. AB - OBJECTIVE: The purpose of this study was to document the usefulness of early triphasic image acquisition during gadolinium-enhanced multiphase dynamic MR imaging for the detection of contrast enhancement in small hepatocellular carcinoma. SUBJECTS AND METHODS: Dynamic MR imaging was performed in 332 patients at 1.5 T using a T1-weighted fast low-angle shot sequence (TR range/TE, 113 130/4.1; flip angle, 80 degrees). With the start of bolus injection of gadolinium dimeglumine (0.1 mmol/kg) into the antecubital vein, three sets of contrast enhanced images of the entire liver were obtained serially at 10, 35, and 60 sec. Contrast enhancement of the tumor detected on each phase was evaluated in conjunction with the unenhanced T1-weighted images for 52 patients with small hepatocellular carcinomas up to 2 cm in diameter (n = 79). RESULTS: The first phase images depicted a larger number (n = 57) of contrast enhancement of small hepatocellular carcinoma tumors than the second (n = 44) or third phase images (n = 14). Twenty-six lesions were enhanced only during the first phase, and nine, during the second phase. The detection rate of contrast enhancement in the combination of first and second phases was the same as the overall detection rate for all three phases (70/79, 89%) and significantly higher than those of each phase. CONCLUSION: Early multiple image acquisition is helpful for the assessment of small hepatocellular carcinoma and is enabled by the high detection rate of contrast enhancement regardless of individual variations in the timing of enhancement. PMID- 10470885 TI - Viral-induced cirrhosis: grading of severity using MR imaging. AB - OBJECTIVE: The purpose of this study was to determine whether MR imaging can be used to grade the severity of cirrhosis. MATERIALS AND METHODS: The MR examinations of 46 patients with cirrhosis were retrospectively reviewed independently by two radiologists and correlated with clinical severity assessed by Child-Pugh classification. MR imaging analysis by reviewers who were unaware of clinical status included comparison of volume indexes (computed as the product of three axis measurements) of the spleen and each segment of the liver, and changes in hepatic contour, iron or fat deposition, and presence of varices and collaterals. RESULTS: Volume index of the spleen and the presence of ascites and varices were significantly and positively correlated (p = .008, .002, .0001, respectively) with the clinical severity of cirrhosis (Child-Pugh classifications), and volume indexes of the posterior, medial, and lateral segments of the liver were significantly and inversely correlated (p = .001, .049, .041, respectively). On an MR scoring system based on four items (volume index of the spleen; volume index of posterior + medial + lateral segments; presence of ascites; and presence of varices and collaterals), averaged total MR scores were 2.5 +/- 0.3, 4.9 +/- 0.6, and 7.9 +/- 0.8 for Child-Pugh grades A, B, and C, respectively (p < .0001). The accuracy of MR scoring in distinguishing between clinical Child-Pugh grade A cirrhosis and further grades was 89%, the sensitivity was 93%, and the specificity was 82%. CONCLUSION: An MR scoring system can be used to grade the severity of cirrhosis. PMID- 10470887 TI - Hyperenhancing focal liver lesions: differential diagnosis with helical CT. PMID- 10470888 TI - Helical CT cholangiography for the detection and localization of bile duct leakage. AB - OBJECTIVE: We describe the use of helical CT cholangiography or helical CT after administration of biliary i.v. contrast material to verify and localize bile duct leakage. CONCLUSION: Helical CT cholangiography revealed bile leaks in seven patients after penetrating trauma or liver or gallbladder surgery and excluded bile leaks in two patients with blunt liver trauma. Endoscopic retrograde cholangiography was performed in only one of seven patients with bile leakage and was avoided in an additional two patients in whom CT cholangiography excluded leakage. The method is a feasible, noninvasive tool for the detection and localization of bile leaks and may help avoid endoscopic retrograde cholangiography. PMID- 10470889 TI - Choledochal cyst: an unusual type of cystic dilatation of the cystic duct. PMID- 10470890 TI - A large peribiliary cyst in the extrahepatic bile duct. PMID- 10470891 TI - Fine-needle transjugular intrahepatic portosystemic shunt procedure with CO2. PMID- 10470892 TI - Transjugular intrahepatic portosystemic shunt placement in a patient with absent right superior vena cava and persistent left superior vena cava. PMID- 10470893 TI - Failure of right portal vein ligation to induce left lobe hypertrophy due to intrahepatic portoportal collaterals: successful treatment with portal vein embolization. PMID- 10470894 TI - Percutaneous abdominal and pelvic interventional procedures using CT fluoroscopy guidance. AB - OBJECTIVE: The purpose of our study was to assess the use of low-milliamperage CT fluoroscopy guidance for percutaneous abdominopelvic biopsy and therapeutic procedures. MATERIALS AND METHODS: We reviewed the clinical records and relevant imaging studies of 97 patients who underwent 119 percutaneous CT fluoroscopy guided abdominal or pelvic procedures: fluid collection aspiration or drainage catheter insertion (n = 59), biopsy (n = 49), hepatocellular carcinoma ethanol ablation (n = 6), chemoneurolysis (n = 4), and brachytherapy catheter insertion (n = 1). These procedures were guided using a helical CT scanner providing real time fluoroscopy reconstruction at six frames per second. A control panel and video monitor beside the gantry allowed direct operator control during all interventional procedures. RESULTS: One hundred twelve (94.1%) procedures were successfully performed using either a stand-off needle holder and continuous real time CT fluoroscopy guidance or incremental manual insertion and intermittent CT fluoroscopy to confirm position. Image quality using low milliamperage was adequate for needle or drainage tube placement in all but two low-contrast liver lesions. Two hematomas were accessed but yielded no fluid on aspiration; one drainage procedure was abandoned after the patient developed endotoxic shock. Imaging of ethanol distribution during injection facilitated tumor ablation and neurolytic procedures. CT fluoroscopy allowed rapid assessment of needle, guidewire, dilator, and catheter placement, especially in nonaxial planes. Average CT fluoroscopy time for biopsy and therapeutic procedures was 133 sec (range, 35-336 sec) and 186 sec (range, 20-660 sec), respectively. CONCLUSION: CT fluoroscopy is a practical clinical tool that facilitates effective performance of percutaneous abdominal and pelvic interventional procedures. PMID- 10470895 TI - MR-guided RF thermal ablation of the kidney in a porcine model. AB - OBJECTIVE: The purpose of the study was to evaluate the ability of MR imaging to reveal RF interstitial thermotherapy in the porcine kidney, as a model for future human trials, and to provide guidance for RF probe insertion. SUBJECTS AND METHODS: Ten MR-guided RF ablations were performed in the kidneys of three pigs. A 17-gauge monopolar RF probe electrode was inserted into the renal cortex using MR guidance, and RF was applied for 10 min. After postprocedure imaging (T2 weighted, turbo short inversion time inversion recovery [STIR], and T1-weighted sequences), the kidneys were harvested for pathologic examination. RESULTS: Successful RF probe placement was accomplished in all cases; the interventional procedure time for probe insertion ranged from 4 to 15 min. The thermal lesion size varied from 7 to 14 mm perpendicular to the probe track and was best seen on turbo STIR images. Turbo STIR had the highest renal cortex-to-RF thermal lesion contrast-to-noise ratio with an average of 4.4 +/- 3.5. Turbo STIR imaging never overestimated pathologic lesion diameter with a mean difference of 1.5 +/- 1.4 mm. Three subcapsular hematomas occurred. which could be detected immediately on postprocedure imaging. CONCLUSION: MR-guided RF thermal ablation in the porcine kidney was found to be feasible, but minor complications occurred. Induced thermal lesion size was best monitored using turbo STIR images. In the future, RF ablation may offer an alternative treatment option for renal cancer. PMID- 10470896 TI - A novel application for the end-fire sonographic probe: guidance during cryoablation of renal masses. PMID- 10470897 TI - Combined sonographic and fluoroscopic guidance: a modified technique for percutaneous sclerosis of low-flow vascular malformations. PMID- 10470898 TI - Assessment of deep venous thrombosis using routine pelvic CT. AB - OBJECTIVE: The purpose of this study was to assess routine pelvic CT both for accuracy in diagnosis of deep venous thrombosis and for frequency of detection of clinically unsuspected pelvic thrombi. MATERIALS AND METHODS: We reviewed the CT records of patients who had undergone pelvic CT during a 6-month period and cross referenced these records to reports on lower extremity venous sonography to identify patients who had undergone this test within 1 week of pelvic CT. We compared the frequency and location of venous thrombosis revealed through these two tests. To identify unsuspected deep venous thrombosis detected on pelvic CT, we reviewed the charts of patients for whom a clot was reported to determine if it had been clinically suspected before CT. RESULTS: Among the 52 patients who underwent both CT and lower extremity venous sonography, the findings were in agreement for 49 (94%). The techniques disagreed for three patients; two patients had deep venous thrombosis detected on sonography but not on CT and the other had a clot detected on CT but not on sonography. The prevalence of unsuspected deep venous thrombosis detected by CT was 1.1%, and 73% of these patients underwent anticoagulation therapy without further confirmatory tests. CONCLUSION: Unsuspected deep venous thrombosis is commonly seen on routine pelvic CT and should be carefully sought during such examinations. CT appears to be relatively accurate in the detection of deep venous thrombosis. Thus, CT venography combined with pulmonary CT angiography may significantly increase the percentage of patients who are appropriately treated for thromboembolic disease because both deep venous thrombosis and pulmonary emboli can be identified by this combined test. PMID- 10470899 TI - Intravascular sonography in the assessment of traumatic injury of the thoracic aorta. AB - OBJECTIVE: The objective of this study is to describe the use of intravascular sonography in the evaluation of suspected injury of the thoracic aorta as an adjunctive tool to digital subtraction arteriography (DSA). SUBJECTS AND METHODS: Images of the thoracic aorta were obtained using DSA and intravascular sonography in 20 consecutive patients who underwent arteriography after chest trauma. A 6 French, 12.5-MHz sonography catheter was used for the intravascular sonography study. Diagnoses based on the review of both studies were compared. RESULTS: Five of 20 patients had aortic or great vessel injuries confirmed by surgery. Intravascular sonographic findings were positive (one false-positive) in seven patients, whereas DSA findings were positive in six patients (one false-positive and one false-negative). Surgery showed the lesion that was false-positive using both techniques to be a ductus diverticulum. Intravascular sonographic findings included intimal flaps, intramural lesions (hematomas), pseudoaneurysms, and perivascular hematomas. CONCLUSION: In our initial limited experience, intravascular sonography of the thoracic aorta and great vessels effectively identified traumatic injuries. Intramural injury without pseudoaneurysm formation or obvious intraluminal flap was visualized by intravascular sonography in one patient but was not detected by DSA. On the other hand, a ductus diverticulum was erroneously interpreted as trauma using both techniques. Intravascular sonography is a relatively new procedure with an obvious learning curve in the interpretation of the findings. An atypical ductus diverticulum may still be mistakenly interpreted as a sign of traumatic injury of the aorta. Familiarity with intravascular sonography in the setting of aortic trauma is necessary for correct interpretation of the images. The sonographic findings offer views of the aorta that are complementary to those of aortography. PMID- 10470900 TI - Sonographic assessment of an inaccessible chemotherapy infusion port. PMID- 10470901 TI - MR imaging of fibrocartilaginous masses arising on the margins of spondylolysis defects. AB - OBJECTIVE: Spondylolysis is reported in up to 7% of the population. An uncommon process that can accompany a pars interarticularis defect is a fibrocartilaginous mass, which can cause impression on the nerve roots and thecal sac. Recognition is important so that the presence may be addressed at the time of surgery. This report describes the MR imaging appearance and clinical significance of a fibrocartilaginous mass in association with spondylolysis. MATERIALS AND METHODS: We reviewed data regarding 336 patients who had lumbar spine imaging at our institution during a 12-month period. Contiguous axial and sagittal MR imaging using T1-weighted and fast spin-echo T2-weighted sequences was used. Images were evaluated for a mass of tissue surrounding the pars defect with MR characteristics of cartilaginous and fibrous low signal intensity on T1-weighted images and low to intermediate signal intensity on T2-weighted images. The position of the fibrocartilaginous mass and its relation to the thecal sac were noted. Surgical correlation between those patients with a fibrocartilaginous mass and those without was examined. RESULTS: Twenty-nine (8.6%) of 336 patients were identified as having a pars interarticularis defect. A fibrocartilaginous mass was present in 26 (90%) of 29 patients. Six (21%) of the 29 patients had a mass effect on the thecal sac. Of the remaining 23 patients, 20 had a fibrocartilaginous mass external to the thecal sac, and three did not show any fibrocartilaginous mass. Histologic analysis in one patient confirmed the fibrous and cartilaginous nature of the lesion at the pars defect. CONCLUSION: A fibrocartilaginous mass was present in 90% of the patients with spondylolysis. Mass effect on the thecal sac that required surgery was identified in 21% of the patients with spondylolysis. Awareness of this mass and proper imaging protocols will enable the radiologist to preoperatively alert the surgeon to its presence. PMID- 10470902 TI - Duplex and color Doppler sonographic findings in active sacroiliitis. AB - OBJECTIVE: The aim of this study was to describe the duplex and color Doppler sonographic findings in active sacroiliitis. SUBJECTS AND METHODS: Forty-one joints in 21 patients with active sacroiliitis, 20 sacroiliac joints in 10 patients with osteoarthritis, and 30 sacroiliac joints of 15 asymptomatic volunteers were investigated on duplex and color Doppler sonography. We investigated whether a vessel was present around the posterior portions of sacroiliac joints with color Doppler sonography. When an artery was detected, the resistive index (RI) was measured using duplex Doppler sonography in all groups and also after treatment in the patients with active sacroiliitis. RESULTS: Vascularization around the posterior portions of sacroiliac joints was seen in 41 joints of the 21 patients with active sacroiliitis, nine joints of six patients with osteoarthritis, and 13 joints of eight volunteers. The mean RI values were 0.62 +/- 0.13, 0.91 +/- 0.09, and 0.97 +/- 0.03, respectively. In the patients with active sacroiliitis, the mean RI value was 0.91 +/- 0.07 after therapy. The RI values for the patients with active sacroiliitis were significantly different from those of the patients with osteoarthritis (p < .001) and of the volunteers (p < .001). In addition, the RI values were significantly different before and after treatment in the patients with active sacroiliitis (p < .001). CONCLUSION: Vascularization around the posterior portions of sacroiliac joints increased and RI values decreased in patients with active sacroiliitis. Color and duplex Doppler sonography were able to reveal these changes and can be used in the diagnosis of active sacroiliitis and follow-up after treatment. Thus, RI values may be a quantitative indicator for clinical symptoms in patients with active sacroiliitis. PMID- 10470903 TI - Sonography in the diagnosis of carpal tunnel syndrome. AB - OBJECTIVE: The few papers published on the use of sonography in carpal tunnel syndrome suggest it may be a useful diagnostic test. This study aims to prospectively evaluate the use of sonographic measurements of the median nerve in the diagnosis of carpal tunnel syndrome. SUBJECTS AND METHODS: Patients with documented carpal tunnel syndrome and a group of asymptomatic control subjects were enrolled and underwent high-resolution sonography of the carpal tunnel. A small-footprint linear array transducer was used to scan and measure the median nerve cross-sectional area and the maximum transverse and anteroposterior diameters. Data from the patient group and the control group were compared to establish optimal diagnostic criteria for carpal tunnel syndrome. RESULTS: Sixty eight carpal tunnel syndrome patients (50 women, 18 men) with 102 affected nerves and 68 nerves in 36 asymptomatic controls (23 women, 13 men) were examined. Qualitative assessment alone was found to be unreliable. All measurements showed significant differences between patients and controls. The most predictive measurement was swelling of the median nerve, which was significantly greater in carpal tunnel syndrome patients compared with controls (mean, 0.13 cm2 versus 0.07 cm2). Thus, quantitative assessment of the median nerve provides an accurate diagnostic test (sensitivity, 82%; specificity, 97%), with an area larger than 0.09 cm2 being highly predictive of carpal tunnel syndrome. CONCLUSION: We confirm that median nerve cross-sectional area measurement correlates well with the presence of carpal tunnel syndrome and is both sensitive and specific for the diagnosis. PMID- 10470904 TI - Anterolateral compartment of the ankle in the lateral impingement syndrome: appearance on CT arthrography. AB - OBJECTIVE: This article describes the appearance of the anterolateral recess of the posttraumatic ankle on CT arthrography and clearly shows the benefit of this technique in the diagnosis of soft-tissue impingement. MATERIALS AND METHODS: The study was carried out on 44 patients examined by CT arthrography and arthroscopy for chronic, posttraumatic ankle pain. The analysis principally concerned the lateral compartment--in particular, the synovial regularity and the cartilage of the talar dome. RESULTS: Four CT patterns were identified: type 0, uniform filling of the recess with clear limits; type I, intraarticular linear structure outlined by contrast agent; type II, nodular formation in the lateral groove; and type III, irregular appearance of the edges of the lateral groove. Ninety-one percent (10/11) of type II patterns were characterized as meniscoid lesions by arthroscopic examination, 100% (14/14) of type III patterns corresponded to an abundant fibrous reaction, and arthroscopic findings were normal for 100% (2/2) of type 0 patterns and 88% (15/17) of type I patterns. The latter was thus considered as a normal variant outlining the accessory anterior inferior tibiofibular ligament. Type II and III lesions were statistically associated (p = .001) with a chondropathy when time from initial trauma was greater than 22 months. CONCLUSION: CT arthrography provides evidence of anterolateral soft tissue impingement--in particular, in type II or III patterns. These lesions are statistically associated with a chondropathy. PMID- 10470905 TI - Quantification of intact quadriceps tendon, quadriceps tendon insertion, and suprapatellar fat pad: MR arthrography, anatomy, and cryosections in the sagittal plane. AB - OBJECTIVE: The purpose of this study was to quantify quadriceps tendon length, thickness, and insertion in relation to the suprapatellar fat pad. SUBJECTS AND METHODS: We used three methods to analyze the anatomy of intact quadriceps tendons and insertions into the patellar base: MR arthrography (53 knees with intact extensor mechanisms), gross anatomy (16 cadaveric knees), and cryosections (four cadaveric knees). With an electronic cursor, two observers independently quantified the extensor mechanism on midline sagittal T1-weighted spin-echo sequences acquired on a low-field-strength (0.23 T) scanner. RESULTS: On MR arthrograms, quadriceps tendon length, determined from the superior patellar pole to the most superior part of the suprapatellar recess, measured 49 +/- 7 mm in women and 50 +/- 9 mm in men. Thickness of quadriceps tendon at three sites (suprapatellar recess, center, and superior patellar pole) measured 7 +/- 1 mm in women and 8 +/- 1 mm in men. Thickness was significantly larger in men at all measurement locations. Quadriceps tendon insertion and the suprapatellar fat pad along the patellar base measured 16 +/- 2 and 6 +/- 2 mm, respectively, in women, and 18 +/- 3 and 7 +/- 2 mm, respectively, in men. CONCLUSION: On midline MR images, sagittal thickness of the quadriceps tendon and its insertion was significantly larger in men than in women. The prevalence of the suprapatellar fat pad was 100%. PMID- 10470906 TI - MR imaging of plantar fasciitis: edema, tears, and occult marrow abnormalities correlated with outcome. AB - OBJECTIVE: We sought to evaluate various MR imaging signs of plantar fasciitis and to determine if a difference in these findings exists between clinically typical and atypical patients with chronic symptoms resistant to conservative treatment. CONCLUSION: We found signs on MR imaging that, to our knowledge, have not been described in the scientific literature for patients with plantar fasciitis. These signs included occult marrow edema and fascial tears. Patients with these manifestations seemed to respond to treatment in a manner similar to that of patients in whom MR imaging revealed more benign findings. PMID- 10470907 TI - Maisonneuve fracture of the fibula: implications for imaging ankle injury. PMID- 10470908 TI - MR imaging of myotendinous strain. AB - Myotendinous strain predominantly exhibits pathognomonic MR features. Whenever a patient's musculoskeletal symptoms lack temporal correlation with exercise, MR imaging may be valuable in distinguishing strain injury from other disorders of the myotendinous unit. Compared with myotendinous strain, myositis ossificans has variable and nonspecific MR appearances that may simulate neoplasm and require biopsy for diagnosis. PMID- 10470909 TI - Phalangeal microgeodic syndrome: findings on MR imaging. PMID- 10470910 TI - Pediatric diffuse lung disease: diagnosis and classification using high resolution CT. AB - OBJECTIVE: Our purpose was to categorize high-resolution CT findings in children with diffuse lung disease and to evaluate the accuracy of diagnoses made using CT. MATERIALS AND METHODS: The chest radiographs and high-resolution CT scans of 20 children (1-16 years old; median, 9 years old) with biopsy-proven chronic diffuse lung diseases were reviewed separately by two independent chest radiologists. Thirteen types of diffuse lung disease were included in the study. Radiographic and CT features were noted, and three choices of diagnosis were recorded, with the confidence level. RESULTS: Diagnoses were made with a high degree of confidence (definite or probable) in 25 of 40 interpretations of CT scans, compared with only five of 40 interpretations of chest radiographs (p < .001). Fourteen (56%) of the 25 confident first-choice diagnoses on CT scans were correct, compared with two (40%) of the five interpretations on chest radiographs. Diseases were classified as belonging to one of five distinct groups on the basis of dominant CT features. Airway disease (n = 5) (bronchiolitis obliterans or bronchocentric granulomatosis) showed geographic hyperlucency on CT. Septal disease (n = 4) (lymphangiomatosis, hemangiomatosis, or microlithiasis) showed septal thickening. Infiltrative lung disease (n = 7) (desquamative interstitial pneumonitis, hypersensitivity pneumonitis, or lymphoid interstitial pneumonitis) showed ground-glass opacity. Air-space disease (n = 3) (aspiration, vasculitis, or bronchiolitis obliterans organizing pneumonia) showed lung consolidation. Langerhans' histiocytosis (n = 1) showed cysts and nodules. Surprisingly little overlap was seen among these groups. CONCLUSION: CT increases the level of diagnostic confidence for pediatric infiltrative lung disease, improves diagnostic accuracy, and provides a useful classification system. PMID- 10470911 TI - Femoral growth lines: bony birthmarks in infants. AB - OBJECTIVE: During a study of infants who were being examined for antenatal renal dilatation, we noted that many had growth lines in their proximal femur on postnatal radiographs. We decided to determine the prevalence of growth lines in healthy infants. MATERIALS AND METHODS: Voiding cystourethrograms of 791 neonates and infants, 0-6 months old, were reviewed. All who had documented severe illness or no reasonable view of the proximal femur were excluded, resulting in a cohort of 633 healthy infants. Each study was coded for presence or absence of a growth line in the proximal femur, and the distance of the growth line from the metaphyseal edge was measured to the nearest 0.5 mm. Radiographs were obtained on a unit with fixed tube-film distance. Data on the mode of delivery were collected for 136 infants. RESULTS: Of 633 eligible infants, 247 (39%) of 633 had a discernible growth line. The distance of the growth line from the metaphysis, in millimeters, correlated significantly with age in days (r = .81, p < .01). Infants delivered vaginally were more likely to have a growth line than were those born by cesarean delivery (p = .049). CONCLUSION: A growth line in the proximal femur is common in healthy infants. The rate of longitudinal growth of the proximal femoral metaphysis, on radiographs, is approximately 1 mm per 11 days (1 mm per 13 days when corrected for magnification). Approximation of the timing of prenatal and postnatal stressful events that result in a growth disturbance line may be possible. PMID- 10470912 TI - Transanal MR imaging after repair of anorectal anomalies in children: appearances in pull-through versus posterior sagittal reconstructions. AB - OBJECTIVE: Our objective was to use transanal MR imaging to compare the anatomic appearance of the components of the anal sphincter and the pattern of scarring after a pull-through perineoplasty versus a posterior sagittal anorectoplasty. SUBJECTS AND METHODS: Sixteen children ranging in age from 10 months to 15 years (mean, 10 years) were imaged using transanal receiver coils appropriate to the size of the child. Five had low, four intermediate, and seven high anomalies. Seven had undergone a transanal pull-through procedure, and nine had undergone posterior sagittal reconstruction. The integrity of the muscles was assessed on T1-weighted and short inversion time inversion recovery transverse and coronal images using a qualitative MR imaging score. The pattern of scarring was also assessed. RESULTS: In the transanal pull-through group, four of seven patients showed external sphincter deficiency. A circumferential low-signal-intensity band was seen inferior to the sphincter in six patients. All posterior sagittal reconstructions had a long posterior midline scar. Five of nine patients showed external sphincter deficiency, whereas a further two had internal sphincter deficiency. No differences were seen in MR imaging scores for each operative procedure for all grades of severity of anorectal anomaly. However, a comparison between high and intermediate anomalies showed a small improvement in MR imaging score using the transanal pull-through procedure (Mann-Whitney U test = 3, p < .03). Manometric pressures obtained in 13 patients were poor. CONCLUSION: Transanal MR imaging identifies focal defects and patterns of scarring of the anal sphincter complex in infants and children and provides valuable information about individual muscle components. PMID- 10470913 TI - Imaging pediatric oncologic emergencies of the abdomen. PMID- 10470914 TI - Contrast-enhanced sonography of vesicoureterorenal reflux in children: preliminary results. AB - OBJECTIVE: The aim of our study was to evaluate contrast-enhanced sonography as an alternative to radiographic voiding cystourethrography in the detection of vesicoureteral reflux. SUBJECTS AND METHODS: A total of 46 children, ranging in age from 3 weeks to 14 years (median: 4 years, 6 months) with 92 ureterorenal units were investigated for reflux using radiographic voiding cystourethrography and contrast-enhanced sonography in one session. After sonography of the urinary tract, the bladder was filled with saline solution via a catheter. Later, a contrast-enhancing agent was instilled and sonography was repeated. Documentation was done using S-VHS video and a laser camera. Reflux was diagnosed when microbubbles were observed in the ureter or in the renal pelvis. In addition, conventional voiding cystourethrography was performed. Patients without micturition during either sonography or radiographic examination were excluded (eight ureterorenal units). RESULTS: The findings obtained by contrast-enhanced sonography and voiding cystourethrography were concordant in 78 ureterorenal units (92.9%). No reflux was detected in 67 units (79.8%) by either method. With voiding cystourethrography as the standard of reference, the sensitivity of contrast-enhanced sonography was 91.7%; the specificity, 93.1%; and the accuracy, 92.9%. The positive predictive value was 68.8%, and the negative predictive value was 98.5%. CONCLUSION: Contrast-enhanced sonography is highly sensitive for the detection of vesicoureteral reflux. Therefore, it may reduce the number of radiographic investigations. PMID- 10470915 TI - Sonographic appearances of extratesticular fluid and fluid-containing scrotal masses in infants and children: clues to diagnosis. PMID- 10470916 TI - Different phases of renal enhancement: role in detecting and characterizing renal masses during helical CT. AB - Although helical CT offers greater flexibility in data acquisition and reconstruction than does conventional axial CT, new opportunities for error must be understood so that optimal protocols are used that minimize the likelihood of misdiagnosis. Most nonrenal abdominal helical CT scans are obtained late in the corticomedullary phase of renal enhancement because of the preferable enhancement of other parenchymal organs. CT evaluation of the kidneys during the corticomedullary phase or at an intermediate phase between the corticomedullary and nephrographic phases has significant limitations. Therefore, dedicated renal CT performed for the detection of suspected renal masses or for the characterization of known renal masses must include images obtained during later phases of enhancement (i.e., nephrographic or excretory phase). Nephrographic or excretory phase images appear to be similar to one another but superior to corticomedullary phase images in the ability to both detect and characterize renal masses. Corticomedullary phase images should always be obtained when information about the renal vasculature is desired or when there is a possibility that a detected renal mass may represent an aneurysm or an arteriovenous malformation or fistula. Optimal technique of helical CT for staging renal cancers may require use of both corticomedullary and nephrographic or excretory phase images, although work in this area is still preliminary. PMID- 10470918 TI - Depiction of renal perfusion defects with contrast-enhanced harmonic sonography in a porcine model. AB - OBJECTIVE: Our purpose was to test the feasibility and optimization of contrast enhanced gray-scale harmonic sonography for the evaluation of focal renal perfusion defects in an animal model. SUBJECTS AND METHODS: We performed arteriography and contrast-enhanced harmonic sonography on six anesthetized piglets after embolization of each kidney with an autologous clot through the main renal artery. Harmonic images were obtained with continuous (30 Hz) and ECG triggered acquisition. The two harmonic sonography strategies were compared with respect to the number and conspicuity of devascularized areas, and correlation was made with arteriographic findings. RESULTS: Contrast-enhanced harmonic sonography showed focal areas of absent or diminished vascularity that corresponded closely with perfusion defects seen on angiography. Enhancement ratios to perfused cortex were significantly higher than to devascularized cortex in both continuous (mean +/- SD, 469 +/- 5% versus 102 +/- 8%, p < .0005 by t test) and triggered (673 +/- 7% versus 198 +/- 7%, p < .0001) modes. Triggered acquisition increased the conspicuity of perfusion defects over that obtained with continuous imaging (p < .002 by t test). CONCLUSION: Contrast-enhanced harmonic sonography is an effective method of depicting focal renal perfusion defects. Triggered acquisition further improves lesion conspicuity. PMID- 10470919 TI - Using Doppler sonography to reveal renal artery stenosis: an evaluation of optimal imaging parameters. AB - OBJECTIVE: The aim of this study was to determine the accuracy of direct and indirect parameters for the diagnosis of renal artery stenosis and to determine the most useful thresholds for these parameters. SUBJECTS AND METHODS: One hundred twenty-five arteries in 63 patients were examined with renal Doppler sonography and angiography for the presence or absence of renal artery stenosis. Arteries were considered stenosed on angiography if there was a diameter reduction of greater than 60%. Renal Doppler sonographic measures of peak systolic velocity, renal aortic ratio, acceleration time, and acceleration were recorded and compared with the angiographically determined presence or absence of disease. RESULTS: Doppler examination was technically successful in 87% of kidneys and 76% of patients. Receiver operating characteristic analysis showed the optimal peak systolic velocity threshold to be 180 cm/sec and the optimal renal aortic ratio threshold to be 3.0. An acceleration time greater than 70 msec and an acceleration less than 300 cm/sec2 yielded sensitivities of 41% and 56%, respectively, and specificities of 85% and 62%, respectively. Combining a renal aortic ratio of greater than 3.0 or peak systolic velocity greater than 180 cm/sec provided the best combination of parameters with a sensitivity and sensitivity at 85% and 76%, respectively. CONCLUSION: The most accurate use of parameters was found to be a combination of either peak systolic velocity greater than 180 cm/sec or renal aortic ratio greater than 3.0. Indirect parameters were not found to be useful in predicting the presence or absence of renal artery stenosis. PMID- 10470920 TI - MR imaging of endometrial stromal sarcoma: correlation with pathologic findings. AB - OBJECTIVE: This study describes MR imaging of eight patients with endometrial stromal sarcoma and correlates the imaging findings with histopathologic findings. To our knowledge, the radiologic findings of this entity have not been reported in the literature. CONCLUSION: Endometrial stromal sarcoma typically presents with extensive myometrial involvement, which is either sharply demarcated or diffusely infiltrative. Bands of low signal intensity are observed within the areas of myometrial involvement on T2-weighted images. These bands correspond to the preserved bundles of myometrium on pathologic examination. Tumor extension along the vessels or ligaments is another characteristic of MR imaging of endometrial stromal sarcoma. PMID- 10470921 TI - Sonographic evaluation for ectopic pregnancy: transabdominal scanning of patients with nondistended urinary bladders as a complement to transvaginal sonography. PMID- 10470922 TI - Wandering wires: frequency of sternal wire abnormalities in patients with sternal dehiscence. AB - OBJECTIVE: The purpose of this study is to assess the frequency of various sternal wire abnormalities on chest radiographs of patients with sternal dehiscence and to determine the role of radiography in detecting or confirming this complication. MATERIALS AND METHODS: We used our computerized hospital information system to identify all patients with a diagnosis of sternal dehiscence from January 1993 through June 1998. Clinical data were obtained by retrospective chart review. A chest radiograph from the date of diagnosis was compared with the first postoperative radiograph obtained after median sternotomy. Each radiograph was retrospectively reviewed by two radiologists who assessed three sternotomy wire abnormalities: displacement (offset of one or more wires in relation to others in the vertical row), rotation (alteration in the axis of a wire compared with its orientation on a baseline radiograph), and disruption (unraveling or fracture of a wire). We also reviewed a series of postoperative radiographs in a group of matched controls who had an uneventful postoperative course with no clinical evidence of dehiscence. RESULTS: The study cohort included 19 patients, 13 men and six women, who were 49-84 years old (mean, 66 years). The chest radiographs revealed sternal wire abnormalities in 17 (89%) of 19 patients with sternal dehiscence, including displacement in 16 (84%) of 19 patients, rotation in 10 (53%) of 19 patients, and disruption in four (21%) of 19 patients. The mean number of displaced wires per patient was 2.3 (range, 1 5). The mean distance of maximal displacement was 20 mm (range, 6-45 mm). Radiographic abnormalities preceded the clinical diagnosis in 13 (68%) of 19 patients. We observed no case of sternal wire displacement, rotation, or disruption in the control group. CONCLUSION: Sternal wire abnormalities, most notably displacement, are present in most patients with sternal dehiscence; radiographic abnormalities precede the clinical diagnosis in most cases. PMID- 10470923 TI - Using urokinase to treat malignant pleural effusions. PMID- 10470924 TI - Gastrointestinal/genitourinary case of the day. Lymphomatous infiltration of the right ureter. PMID- 10470925 TI - Gastrointestinal/genitourinary case of the day. Incarcerated inguinal hernia of the left fallopian tube and ovary. PMID- 10470926 TI - Gastrointestinal/genitourinary case of the day. Liver infarction in a patient with sickle cell anemia, splenic atrophy, and gallstones. PMID- 10470927 TI - Gastrointestinal/genitourinary case of the day. von Hippel-Lindau disease. PMID- 10470928 TI - Musculoskeletal case of the day. Clear cell chondrosarcoma. PMID- 10470929 TI - Musculoskeletal case of the day. Multiple myeloma. PMID- 10470930 TI - Musculoskeletal case of the day. Mazabraud's syndrome (intramuscular myxomas associated with fibrous dysplasia of bone). PMID- 10470931 TI - Musculoskeletal case of the day. Tuberculoid leprosy. PMID- 10470932 TI - Neuroradiology case of the day. Kaposi's sarcoma of the nasopharynx, oropharynx, and hypopharynx. PMID- 10470933 TI - Neuroradiology case of the day. Polymorphous low-grade adenocarcinoma of a minor salivary gland in the right buccal space. PMID- 10470934 TI - Neuroradiology case of the day. Ganglioglioma. PMID- 10470935 TI - Neuroradiology case of the day. Spinal cord compression caused by extramedullary hematopoiesis. PMID- 10470936 TI - Pediatric case of the day. Tuberculosis dactylitis and primary pulmonary tuberculosis. PMID- 10470937 TI - Pediatric case of the day. Sacrococcygeal teratoma, type IV. PMID- 10470938 TI - Pediatric case of the day. Congenital pulmonary arteriovenous malformation. PMID- 10470939 TI - Pediatric case of the day. Total colonic Hirschsprung's disease. PMID- 10470940 TI - Thoracic case of the day. Thoracic myelolipomatosis. PMID- 10470941 TI - Thoracic case of the day. Mounier-Kuhn syndrome (tracheobronchomegaly). PMID- 10470942 TI - Thoracic case of the day. Kartagener's syndrome. PMID- 10470943 TI - Thoracic case of the day. Primary diffuse tracheobronchial amyloidosis. PMID- 10470944 TI - Vascular and interventional case of the day. Peristomal varices. PMID- 10470945 TI - Vascular and interventional case of the day. Pancreatic head pseudoaneurysm treated with percutaneous thrombin injection. PMID- 10470946 TI - Vascular and interventional case of the day. Arteriovenous fistula in a transplanted kidney. PMID- 10470947 TI - Vascular and interventional case of the day. Infected vascular grafts from occult sternal osteomyelitis. PMID- 10470948 TI - Needle choice for sonographically guided core biopsy. PMID- 10470949 TI - Transvaginal sonography versus transabdominal pelvic sonography. PMID- 10470950 TI - Multiple stents in treatment of obstructive jaundice associated with Klatskin's tumor. PMID- 10470951 TI - Comparison of interpretation of digitized images and screen-film radiographs. PMID- 10470952 TI - Avulsion fracture of the sublime (coronoid) tubercle of the ulna in throwing athletes. PMID- 10470953 TI - What's in a name--revisited. PMID- 10470954 TI - The glenolabral articular disruption lesion: similarities to the glenoid osteochondral defect. PMID- 10470955 TI - Comparing single-shot RARE and multislice HASTE sequences. PMID- 10470957 TI - Keep the tradition of professional courtesy alive. PMID- 10470958 TI - Bone densitometry, estrogen, and breast imaging. PMID- 10470959 TI - Calcaneal avulsion fractures. PMID- 10470960 TI - Contingency plans for Y2K problems. PMID- 10470961 TI - Notice of duplicate publication, AJR 1997;168:207-212. PMID- 10470962 TI - Notice of duplicate publication, AJR 1998;171:1643-1644. PMID- 10470963 TI - Hepatic vein obstruction in a case of focal nodular hyperplasia. PMID- 10470964 TI - Hemangioblastoma of the filum terminale: MR imaging. PMID- 10470965 TI - Epidermoid inclusion cysts seen on CT of a patient with Gardner's syndrome. PMID- 10470966 TI - Gastric hemangioma associated with phleboliths: CT appearance. PMID- 10470967 TI - Simultaneous identification of medial and lateral bucket-handle tears: the Jack and Jill lesion. PMID- 10470968 TI - CT and MR appearance of Castleman's disease of the neck. PMID- 10470969 TI - Alcohol craving--a renaissance. PMID- 10470970 TI - Psychometric properties of the Penn Alcohol Craving Scale. AB - BACKGROUND: This study introduces the Penn Alcohol Craving Scale (PACS), which has been used in several clinical trials at the University of Pennsylvania's Treatment Research Center. The PACS is a five-item, self-report measure that includes questions about the frequency, intensity, and duration of craving, the ability to resist drinking, and asks for an overall rating of craving for alcohol for the previous week. Each question is scaled from 0 to 6. METHODS: To examine the questionnaire's psychometric properties, we sampled responses from 147 individuals participating in a 9-month combined natrexone (100 mg/day)/psychotherapy trial. The psychotherapy consisted of weekly sessions of nurse-administered medication compliance and supportive treatment. RESULTS: The PACS proved to have excellent internal consistency. Predictive validity was demonstrated via a logistic regression analysis of craving during the 2nd week of the study on alcohol relapse during weeks 3-12 of the trial. Construct validity of the PACS was demonstrated via its convergence with two commonly used measures for assessing craving, the Obsessive Compulsive Drinking Scale and the Alcohol Urge Questionnaire. Lack of correlation between PACS scores and several other noncraving, self-report measures indicates that the PACS also had good discriminant validity. Additional analyses revealed that there were significant differences in craving scores during the initial 3 weeks of the trial among those who did and those who did not relapse during weeks 3-12. CONCLUSION: The PACS is a reliable and valid measure of alcohol craving and can predict which individuals are at risk for subsequent relapse. PMID- 10470971 TI - Differences in the opioid system in selected brain regions of alcohol-preferring and alcohol-nonpreferring rats. AB - BACKGROUND: Previous studies have suggested that alcohol-reinforcing effects are mediated by the endogenous opioid system, which, in turn, stimulates mesolimbic dopaminergic neurotransmission. In addition, evidence obtained in both humans and rats indicates that genetic factors may influence alcohol-drinking behavior. In the present study, we examined several components of the opioid system in selected brain regions of rats bred selectively for their innate alcohol preference (Sardinian preferring = sP) or alcohol aversion (Sardinian nonpreferring = sNP). METHODS: To evaluate whether differences observed were consequent to alcohol intake, sP rats were divided into two subgroups, ethanol naive sP (sP) and ethanol-experienced sP (sPexp). Opioid receptors were labeled, using [3H]naloxone (mu, delta, and kappa receptors), [D-Ala2,N-Me Phe4,Gly,ol5]enkephalin ([3H]DAMGO; mu receptors), and [D-Ala2,D-Leu5]enkephalin ([3H]DADLE; delta receptors), by means of quantitative autoradiography. Enkephalin and dynorphin mRNA contents were measured by in situ hybridization by using 25- and 47-base oligonucleotide probes with sequences complementary to mRNA encoding rat enkephalin or dynorphin. RESULTS: Our results revealed a significant reduction of opioid receptors in caudate-putamen nucleus and in the shell portion of the nucleus accumbens in sP compared with sNP rats. Alcohol intake partially reversed this reduction in the caudate-putamen nucleus. In addition, enkephalin mRNA expression was found to be decreased in the ventral part of caudate-putamen nucleus and increased in the cerebral cortex of sP rats compared with sNP rats; no significant differences were found in dynorphin mRNA expression in any of the brain areas examined. CONCLUSIONS AND SIGNIFICANCE: Differences observed between the two lines of rats may implicate that genetic modifications in the opioid system are possibly responsible for the innate preference of sP rats toward alcohol intake. At the same time, it cannot be excluded that other functions might also be affected to some degree. PMID- 10470972 TI - Chronic alcoholism is associated with an imbalanced production of Th-1/Th-2 cytokines by peripheral blood T cells. AB - BACKGROUND: In the present study, we analyzed, at the intracellular level, the pattern of cytokine secretion by the major CD4+ and CD8strong+ peripheral blood (PB) T-cell subsets in patients with chronic alcoholism, and we correlated it both with the ethanol (EtOH) intake status and with the presence or not of alcoholic liver disease. METHODS: For that purpose, a total of 30 chronic alcoholic patients, 10 without liver disease (AWLD group) and 20 diagnosed with alcoholic liver cirrhosis (ALC) were studied. In all cases, flow cytometric measurement of intracellular expression of interferon-gamma (IFN-gamma), interleukin (IL)-2, and IL-4 was performed on PB CD4+ and CD8strong+ T lymphocytes. RESULTS: After studying AWLD patients, we found increased numbers of both CD4+ and CD8strong+ PB T cells with detectable cytoplasmic levels of the IL 2 and IFN-gamma T helper (Th)-1-associated cytokines, the greater increase being observed for this latter cytokine (p<0.001 for CD4+ and p<0.01 for CD8strong+ T cells). Regarding ALC patients, the pattern of expression of intracellular cytokines by PB T cells was different depending on the status of EtOH intake at the moment of entering this study. Accordingly, as in AWLD patients, ALC individuals who were actively drinking also displayed increased numbers of both CD4+ and CD8strong+ T cells expressing Th-1-associated cytokines. However, in these patients, expression of IFN-gamma, although being significantly greater than that observed in control individuals (p<0.05), was significantly lower than that in AWLD patients (p<0.01 and p<0.05, for CD4+ and CD8strong+ T cells, respectively). After a withdrawal period of > or =1 yr, ALC patients did not show significant changes in the cytoplasmic expression of Th-1-associated cytokines compared with the control group; in contrast, these patients showed a marked increase on the proportion of CD4+ and CD8strong+ T cells expressing IL-4, a Th-2 associated cytokine (p<0.01). After considering the ratio between the number of T cells expressing Th- (IFN-gamma)- and Th-2 (IL-4)-associated cytokines in each individual, we found that there was a significant imbalance in this ratio, with a predominance of IFN-gamma-producing T cells over IL-4+ T lymphocytes during EtOH intake. CONCLUSIONS: Our results showed that in patients with chronic alcoholism, active EtOH intake is associated with a Th-1 pattern of cytokine production by PB T cells. PMID- 10470973 TI - Association of low-voltage alpha EEG with a subtype of alcohol use disorders. AB - BACKGROUND: Neurophysiological traits may identify more homogeneous subgroups of alcoholics. Such discoveries could yield information regarding pathophysiological development, leading to more specific preventive measures and treatments. In an earlier study of 127 individuals, 59 of whom were unrelated, we found that a heritable resting Electroencephalographic (EEG) phenotype, i.e., the low-voltage alpha (LVA) trait, was associated with alcohol use disorders and anxiety disorders. METHODS: We evaluated these findings using an independent, similarly established, dataset of 120 subjects. We also extended the study to a larger set of 149 unrelated individuals from a total sample of 247 subjects for whom psychiatric diagnoses and resting EEG phenotypes were available. Blind-rated psychiatric diagnoses were formulated according to DSM-III-R criteria. RESULTS: In the replication sample, the LVA trait was again more common among subjects with anxiety disorders than among those without. In the total group of unrelated individuals, alcoholics were significantly (3 times) more likely to show the LVA trait than were nonalcoholics. Again, individuals with anxiety disorders were significantly (3 times) more likely to exhibit the LVA trait than were those without anxiety disorders. Of 11 unrelated alcoholics with anxiety disorders, seven showed the LVA trait. It was specifically the LVA trait and not low amplitude alpha activity that was associated with alcohol use disorders. CONCLUSIONS: The results of this replication study and the analysis of the total sample of unrelated individuals support an association between LVA EEG and the subtype of alcohol use disorders associated with anxiety disorders. The LVA phenotype may be a vulnerability factor for alcohol use disorders and anxiety disorders. PMID- 10470974 TI - A preliminary study of acute responses to clamped alcohol concentration and family history of alcoholism. AB - BACKGROUND: The clamping method of alcohol administration was combined with a battery of dependent measures of frontal lobe brain function, and a novel index of acute adaptation, in a preliminary study in order to explore the paradigm's sensitivity to a familial history of alcoholism (FHA). METHODS: Ten family history-positive (FHP) and 10 family history-negative (FHN) adult social drinkers of both genders underwent alcohol clamping. Twenty minutes after the start of an intravenous infusion of alcohol, the breath alcohol concentration was clamped at a target of 60+/-5 mg/dl for 150 min. Initial and adaptive responses to alcohol were assessed using scalar indices of change. One index assessed initial improvements or impairments in brain function after alcohol. The other index assessed acute adaptation (tolerance or sensitization) to alcohol while the brain's exposure to alcohol was held constant. The battery of dependent measures included subjective perceptions, neuropsychological tests, saccadic eye-movement tasks, and event-related potential (ERP) tasks. Effect sizes for FHA were estimated for 10 dependent variables that showed adequate baseline test-retest reliability (r>0.6). RESULTS: FHP subjects showed less intense initial responses to alcohol in subjective perceptions, but greater changes in the latency of volitional saccades and ERP P3 components than did the FHN controls. FHP subjects generally showed greater acute tolerance to alcohol than did controls, who showed more instances of acute sensitization at this moderate breath alcohol concentration. Effect sizes for FHA exceeded 0.4 in more than half of the indices. CONCLUSIONS: The BrAC clamping paradigm assesses initial and adaptive responses of a battery of behavioral and electrophysiological measures of frontal lobe function to ethanol that appear both reliable and sensitive to FHA. PMID- 10470975 TI - Blood ethanol concentrations in rats drinking sucrose/ethanol solutions. AB - BACKGROUND: The addition of sucrose to ethanol solutions results in a substantial increase in ethanol self-administration by rats that are deprived of neither food nor water. However, if sucrose alters ethanol absorption or metabolism, resulting in blood ethanol concentrations (BECs) not different from those resulting from lower intakes of ethanol/water solutions, then the usefulness of sucrose/ethanol mixtures in increasing ethanol consumption is questionable. The present study was conducted to determine whether the addition of sucrose to ethanol solutions altered BECs in an operant self-administration paradigm. METHODS: Tail blood (from male Long-Evans rats) was collected 30 min after the intake of four different solutions, i.e., 5% sucrose/20% ethanol, 5% sucrose/10% ethanol, 2% sucrose/10% ethanol, and 10% ethanol. RESULTS: Ethanol intakes (mean, 1.57+/-0.21 g/kg) and BECs (mean, 78.4+/-9.3 mg/100 ml) were highest when 5% sucrose was added to the ethanol solution. Moreover, the ratios between ethanol intakes and resulting BECs were approximately the same for all solutions. CONCLUSIONS: These findings indicate that, under the conditions of this procedure, the BEC reached is dependent on the amount of ethanol consumed and is not influenced by the addition of sucrose to the solution. PMID- 10470976 TI - CSF monoamine metabolite and beta endorphin levels in recently detoxified alcoholics and healthy controls: prediction of alcohol cue-induced craving? AB - BACKGROUND: Abnormalities in central neurotransmitter systems have been described in alcohol-dependent individuals and may contribute to alcohol craving. This study compared cerebrospinal fluid (CSF) levels of monoamine metabolites and beta endorphin levels in samples from early-onset alcohol-dependent patients (n = 20), late-onset alcohol-dependent patients (n = 14), and healthy controls (n = 23). It also evaluated whether these CSF measures levels predicted the degree of craving experienced in response to an alcohol cue. METHODS: Individuals meeting DSM-III and -IV R-criteria for alcohol dependence, 1 to 3 months postdetoxification, and healthy controls underwent a lumbar puncture. Patients also completed a cue exposure test day between 3 and 15 days later. RESULTS: Alcohol-dependent patients had lower CSF levels of the norepinephrine metabolite MHPG compared with the healthy subjects, but this difference disappeared when differences in age between the groups were accounted for. No other group comparisons between patients and healthy subjects reached significance. CSF levels of the dopamine metabolite HVA were significantly higher in the early-onset patients compared with the late-onset patients and controls. The CSF measures did not predict the precue levels of craving, or the increase in craving after alcohol cue exposure. CONCLUSIONS: These results are inconclusive about the role of monoaminergic dysregulation in recovering alcoholics. They also question the utility of these CSF measures to predict alcohol cue reactivity in patients who have been sober at least 1 month. PMID- 10470977 TI - Alcohol increases commission error rates for a continuous performance test. AB - BACKGROUND: Studying the effects of alcohol on Continuous Performance Test (CPT) performance was of interest for two reasons, i.e., (1) perhaps because of the ease of the task used in previous experiments, alcohol has not been found to impair performance, and (2) CPT commission errors (described below) have been related to impulsive behavior. METHODS: In this study, the CPT featured both an Immediate Memory Task (IMT) and a more difficult Delayed Memory Task (DMT). We compared the performance of 18 subjects under both alcohol and placebo conditions, using a within-subject design. Both the IMT (0.5-sec delay) and the DMT (3.5-sec delay, with distracter stimuli at 0.5-sec intervals) required the subject to respond if a briefly displayed number was identical to the one presented before it. Stimuli included target (identical match), catch (4 of 5 digits matched), and novel (random number) stimuli. On 2 separate days, subjects performed between administrations of three hourly placebo drinks or three hourly drinks containing 0.20 g/kg of alcohol (producing peak breath alcohol concentrations of approximately 0.035%). RESULTS: The main finding was that alcohol consumption increased responses to catch stimuli (i.e., commission errors) in the DMT. In contrast, performance in the IMT (the easier task) was unaffected by alcohol. Commission errors measured during peak breath alcohol concentrations were significantly correlated with scores on the Barratt Impulsivity Scale for both the IMT and DMT. Discriminability (A') between target and catch stimuli was reduced by alcohol for the DMT only. CONCLUSIONS: These data indicate that even small amounts of alcohol can produce measurable changes in CPT performance parameters if the task is of sufficient difficulty and that commission errors can be increased by alcohol consumption. PMID- 10470978 TI - Oral ethanol self-administration in rhesus monkeys: behavioral and neurochemical correlates. AB - BACKGROUND: Previous research has revealed that orally administered ethanol serves as a reinforcer in nonhuman primates. The purposes of the present study were to examine the relationship between ethanol preferences and intakes in two distinct self-administration contexts and to reveal some of the behavioral and neurochemical correlates of oral ethanol self-administration in monkeys. METHODS: Three cohorts of 13 to 29 rhesus monkeys (Macaca mulatta) were socially housed and given daily, 1-hr, one-spout access to an ethanol solution (8.4%, w/v) sweetened with aspartame. Twelve of these monkeys were subsequently selected, individually housed, and given daily, 2-hr, two-spout access to a range of ethanol concentrations (0.25-16%, w/v) concurrently with water. RESULTS: These monkeys (National Institute on Alcohol Abuse and Alcoholism group) showed a marked preference for ethanol (0.5-4%, w/v) over water, and ethanol preferences were 3-fold greater than those of a second group of 12 monkeys (University of Michigan group) purchased from a commercial vendor. Ethanol consumption was consistent across the self-administration paradigms. Monkeys that consumed large quantities of ethanol under the one-spout, social-housing conditions continued to drink large quantities of ethanol under the two-spout, individual-housing conditions (r = 0.86). An association between ethanol preferences and intakes was also demonstrated. Monkeys with the greatest preferences for ethanol over water under the two-spout choice conditions consumed the largest quantities of ethanol (r = 0.82). Finally, cerebrospinal fluid 5-hydroxyindoleacetic acid concentrations were inversely related to ethanol preference but not to ethanol intake. CONCLUSIONS: These results indicate that ethanol consumption is stable across contexts and is positively correlated with the preference for ethanol over water. PMID- 10470979 TI - Serotonergic neuronal activity in the dorsal raphe nucleus of selectively bred alcohol-preferring and alcohol-nonpreferring rats and unselected wistar rats. AB - BACKGROUND: Selectively-bred alcohol-preferring (P) rats have fewer serotonin (5 HT) neurons in the dorsal raphe nucleus (DRN) than do alcohol-nonpreferring (NP) rats. The present study was designed to test the hypothesis that the remaining 5 HT neurons in P rats compensated for their reduced number by increasing neuronal activity. METHODS: Spontaneous activity was recorded from single-spiking and bursting 5-HT neurons in the DRN of unanesthetized paralyzed, alcohol-naive P, NP, and Wistar rats. Firing frequencies, the percentages of action potentials in bursts, and the percentages of bursting neurons were evaluated. RESULTS: There were no significant differences among the three groups of rats in any of the parameters measured. Power analyses were performed on preliminary data to determine the sample sizes necessary for detection of significant differences. The mean firing frequencies of single-spiking 5-HT neurons averaged 1.8 (37 neurons), 1.7 (17 neurons), and 1.8 (41 neurons) spikes per second in P, NP, and Wistar rats, respectively. For bursting 5-HT neurons, the percentages of action potentials in bursts for P, NP, and Wistar rats were 55.0% (24 neurons), 49.7% (18 neurons), and 55.1% (21 neurons). The mean percentages of bursting 5-HT neurons encountered per electrode penetration were 44% for P rats (n = 28), 44% for NP rats (n = 14), and 34% for Wistar rats (n = 26). CONCLUSIONS: The results indicate that the sample of 5-HT neurons recorded in the DRN of P rats had not compensated for a reduced number by altering neuronal activity. PMID- 10470980 TI - A new model of ethanol self-administration in newborn rats: gender effects on ethanol ingestion through a surrogate nipple. AB - BACKGROUND: Ethanol intake in the context of suckling may have distinct and potentially long-lasting consequences for further responsiveness to and acceptance of ethanol, compared to other, more indirect and less natural ways of ethanol exposure early in ontogeny. METHOD: Our findings presented in this paper show that a surrogate nipple technique can be used for the study of early ethanol intake in cesarean-derived rat pups tested before any suckling experience. RESULTS: Neonatal rats attached to and voluntary ingested ethanol through the surrogate nipple as early as 4 hr after birth. Moderate concentrations of ethanol (2% and 5%, v/v) promoted substantial initial suckling behavior, including sustained attachment to the nipple. Higher concentrations (10% and 15%) were not effective in sustaining suckling. Females responded less positively to 10% ethanol than did males. High concentrations of ethanol were less effective in eliciting suckling behavior, probably due to the aversiveness of ethanol odor. However, when ethanol was presented in solution with milk, newborn pups attached to the nipple and ingested even 15% ethanol. Contamination of milk with 15% ethanol was more aversive for females than for males. Newborn rat pups demonstrated similar patterns of nipple attachment and ingestive behavior for 5% ethanol and milk. Initial experience with milk in the context of suckling did not prevent further voluntary ethanol ingestion from the same nipple; furthermore, initial exposure to 5% ethanol did not impair subsequent responsiveness to milk. CONCLUSIONS: Gender differences in responsiveness or sensitivity to ethanol can be detected in rat pups as early as a few hours after birth. The results suggest a leftward shift in the dose-response curve for females compared with males, indicating that female neonates are more sensitive or more responsive than males to ethanol. The similarity of suckling behaviors produced by moderate concentrations of ethanol and milk suggests a certain unity in their reinforcing mechanisms in the context of the first suckling episode. PMID- 10470981 TI - Clozapine's effects on ethanol's motivational properties. AB - BACKGROUND: Although dopamine D1 and D2 receptors have been implicated in ethanol's motivational effects, little is known about the contribution of dopamine D4 receptors. The present experiments examined the effects of clozapine, a dopamine D4 receptor antagonist, on ethanol's aversive, rewarding, stimulant, and reinforcing properties. METHODS: For taste conditioning, adult male Swiss Webster mice received five conditioning trials consisting of 1-hr access to 0.2 M NaCl. After NaCl access on trials 1-4, subjects received clozapine (0, 1, or 2 mg/kg) followed 30 min later by 0, 2, or 4 g/kg ethanol. For place conditioning, Swiss-Webster mice received six pairings of a tactile stimulus with ethanol (2 g/kg, intraperitoneally), clozapine (1 mg/kg, intraperitoneally) + ethanol, or clozapine alone. Locomotor activity in a 30-min test was determined in Swiss Webster mice receiving 0, 0.5, or 1.0 mg/kg clozapine and 0, 1, or 2 g/kg ethanol. In a drinking study, separate groups of adult male C57BL/6J mice were allowed 30-min access to either 10% v/v ethanol mixed in 10% w/v sucrose or 10% sucrose without ethanol. During testing, both groups were given 0 or 1 mg/kg clozapine 30 min before fluid access. RESULTS: Ethanol flavor pairings during taste conditioning reduced subsequent flavor intakes, indicating the development of conditioned taste aversion. Clozapine reduced the magnitude of 4 g/kg ethanol conditioned aversion only on trial 4 at the 2 mg/kg dose. Conditioned place preference for the ethanol-paired stimulus was not altered by clozapine. Clozapine alone did not produce either conditioned preference or aversion. Ethanol-stimulated activity was reduced by clozapine treatment. However, clozapine alone did not alter locomotor activity levels. Clozapine reduced sucrose consumption but did not alter ethanol/sucrose intake. CONCLUSIONS: These data suggest that clozapine influences a limited range of ethanol-motivated behaviors. Specifically, dopamine D4 receptors appear important for ethanol's stimulant effect and possibly ethanol aversion, but not ethanol reward and reinforcement. PMID- 10470983 TI - Individual differences in auditory and visual attention among fetal alcohol affected adults. AB - BACKGROUND: Deficits in attention are commonly identified among patients who have been prenatally exposed to alcohol, and they often affect the ability of the patients to function appropriately in society. METHODS: Eleven adult patients with fetal alcohol syndrome (FAS) or fetal alcohol effects (FAE) were compared with nine adult subjects with no history of prenatal alcohol exposure, using four tests of visually and auditorially mediated attention. RESULTS: In relation to the comparison group, patients with FAS/FAE exhibited substantial deficits in both auditory and visual attention; the auditory deficits were greater. We observed two predominant patterns of deficits among patients with FAS/FAE, one involving both auditory and visual attention problems and the other involving less severe auditory problems and even fewer visual problems. Most subjects with FAS or FAE had some manifestations of attention problems in at least one of the tests of attention used in this study. We present a new graphical representation of individual auditory Continuous Performance Test data across a 6-min period, which, compared with conventional scores, more clearly reveals the markedly disrupted and variable attention patterns displayed by some individuals with FAS or FAE. CONCLUSIONS: These findings suggest that detailed analysis of the pattern of individual performance for each subject is an important aspect of Continuous Performance Test assessment. Our findings further suggest that intellectual performance (intelligence quotient score) alone is not sufficient to account for the patterns of disrupted attention for individuals with FAS/FAE. Assessment of individuals with FAS or FAE should include measurement of attentional functioning in both the visual and auditory modalities. PMID- 10470982 TI - Naltrexone's effect on cue-elicited craving among alcoholics in treatment. AB - BACKGROUND: Advancing knowledge of biobehavioral effects of interventions can result in improved treatments. Thus, a standardized laboratory cue reactivity assessment has been developed and validated to assess the cognitive and psychophysiological responses to a simulated high-risk situation: alcohol cues. The present study investigates the effects of a pharmacotherapy (naltrexone) on a laboratory-based, cue-elicited urge to drink among abstinent alcoholics in treatment. METHODS: Alcohol-dependent subjects were randomized to 12 weeks of naltrexone or placebo after completing a partial hospital program. After approximately 1 week on medication, all received cue reactivity assessment. RESULTS: Significantly fewer patients taking naltrexone reported any urge to drink during alcohol exposure than did those on placebo. Those with any urges reported no decrement in level of the urges. Mean arterial pressure decreased significantly for those on placebo, but not for those on naltrexone, whereas cue elicited decreases in heart rate were not affected by the medication. CONCLUSIONS: The results have implications for models of relapse and naltrexone's effects. Cue reactivity methodology has utility for investigating hypothesized mediators of therapeutic effects of pharmacotherapies as well as behavioral treatments. PMID- 10470984 TI - Effects of ethanol on intestinal absorption of drugs: in situ studies with ciprofloxacin analogs in acute and chronic alcohol-fed rats. AB - BACKGROUND: Previous work from our laboratory on the effect, in rats, of chronic ethanol intake on the intestinal absorption of ciprofloxacin analogs suggested an increased polarity of the lipoidal membrane constituents without effects on the aqueous environment. The aim of the present study was to investigate the influence of acute ethanol intake on the absorption of the same series of compounds. METHODS: The effects of in situ ethanol exposure on intestinal absorption were determined in rats fed either a standard liquid diet or a 5% (w/v) ethanol-containing liquid diet. Acute intestinal exposure to 5% (w/v) ethanol was performed in situ in each feeding group. The biophysical absorption model was used to establish correlations between the actual absorption rate constants, and the lipophilicity indexes, for each group of rats. RESULTS: Acute exposure to ethanol produces an increase only in the absorption of hydrophilic homologs in both control and chronic ethanol fed groups. This suggests the absence of homeoviscous adaptation of the intestinal membrane. The biophysical model used allows us to discriminate between the effects of acute and chronic ethanol treatment on the intestinal membrane. CONCLUSIONS: These results suggest that in contrast to previous reports chronic ethanol treatment increases membrane polarity and acute alcohol intake appears to modify membrane fluidity. PMID- 10470985 TI - Role of yeasts in the salivary acetaldehyde production from ethanol among risk groups for ethanol-associated oral cavity cancer. AB - BACKGROUND: Acetaldehyde, the first metabolite of alcohol, has been proposed to be the carcinogenic substance behind ethanol-related oral cancers. High levels of acetaldehyde are formed from ethanol in saliva by the oral flora, but so far the role of certain microbial species responsible for this phenomenon is not known. Yeasts are common commensals of the oral cavity that have alcohol-oxidizing enzymes, thus providing a potential source of acetaldehyde from ethanol. The aim of this study was to examine the contribution of oral yeasts to the production of ethanol-derived acetaldehyde in the oral cavity. METHODS: Fifty-five saliva samples were divided into two groups, high and low, based on the in vitro salivary acetaldehyde production capacity from ethanol. Yeasts were isolated and identified from these samples, and their acetaldehyde production capacity was determined gas chromatographically by incubating intact cells with ethanol at the physiological pH of 7.4. RESULTS: Yeast colonization was found in 78% of the high acetaldehyde-producing salivas, compared with 47% in the low acetaldehyde producing salivas (p = 0.026). Among carriers, the density of yeasts was higher in the high than in low acetaldehyde producers (p = 0.025). Candida albicans was the main species isolated (88% of all oral isolates). Moreover, C. albicans strains isolated from the high acetaldehyde-producing salivas formed significantly higher acetaldehyde levels from ethanol than C. albicans strains from low-acetaldehyde-producing salivas (73.1 nmol ach/10e6 colony-forming units vs. 43.2 nmol ach/10e6 colony-forming units, p = 0.035). CONCLUSIONS: This study shows that some C. albicans strains have a marked capacity to produce toxic and carcinogenic acetaldehyde from ethanol in vitro. Because the in vitro production of salivary acetaldehyde has been previously shown to correlate with in vivo acetaldehyde production, our finding could be an important microbial pathogenetic factor underlying cancer of the oral cavity associated with ethanol drinking. PMID- 10470986 TI - Effect of alcohol consumption on adult bone mineral density and bone turnover markers. PMID- 10470987 TI - Effects of a DA2/alpha2 agonist and a beta1-blocker in combination with an ACE inhibitor on adrenergic activity and left ventricular remodeling in an experimental model of left ventricular dysfunction after coronary artery occlusion. AB - Renin-angiotensin-aldosterone and sympathetic nervous systems overactivity play a major role in worsening the extent of heart failure. Attenuation of neurohumoral activation with angiotensin-converting enzyme (ACE) inhibitors and beta-blockers has proven beneficial in congestive heart failure. Because ACE inhibition is a recommended treatment for heart failure, this study was designed to test the effects on neurohumoral activation, hemodynamics, and left ventricular (LV) volume of the combination of an ACE inhibitor (delapril) with a DA2-dopaminergic receptor/alpha2-adrenoceptor agonist (CHF-1024) or a beta1-adrenoceptor antagonist (metoprolol) after a moderate to large myocardial infarction (MI) in rats. MI was induced by left coronary artery ligation in 134 rats, and six were not operated on. After 2 months, the animals with ECG evidence of MI were treated for 1 more month with CHF- 1024, 0.33 mg/kg/day or with metoprolol (10 mg/kg/day), delivered through implanted osmotic minipumps, in addition to delapril (6 mg/kg/day) in the drinking water. Daily urinary excretion of norepinephrine (NE) and circulating concentration were measured. Hemodynamic variables were measured, and three-dimensional morphometric analysis was done on the diastole-arrested hearts to quantify infarct size and LV geometry. In conscious animals, delapril alone or with CHF-1024 or metropolol did not modify heart rate or systolic blood pressure. Both combination treatments, however, significantly reduced heart rate in anesthetized animals compared with the group receiving vehicle. Infarct size was not different between treatments, averaging 20-22% of LV volume. The threefold increase of LV chamber volume in infarcted rats was significantly attenuated by delapril alone or with CHF-1024 or metoprolol (-37 to -44%, p<0.05). Treatment with a combination of the ACEi and CHF-1024 tended to normalize the shape of the LV cavity. Urinary NE excretion was unaffected by delapril alone but was reduced by the addition of CHF-1024 or metoprolol. In conclusion, 1 month of treatment with doses of delapril having no hemodynamic effect, reduced LV volume in a model of chronic heart failure. When CHF-1024 or metoprolol was given with delapril, sympathetic activation decreased with no unwanted effects, such as excessive hypotension. PMID- 10470988 TI - Distribution of endothelin receptors in saphenous veins of African Americans: implications of racial differences. AB - The relative density of endothelin-receptor subtypes A and B (ET(A) and ET(B), respectively) on endothelial and smooth muscle cells is the major determinant of the contractile response to endothelin-1 (ET-1). To investigate the effects of race on the distribution of ET receptors, the endothelin-receptor subtypes on membrane fractions prepared from saphenous veins obtained from African-American patients undergoing coronary bypass surgery were analyzed. Similar studies were repeated with endothelium-denuded samples to study the role of endothelium- and smooth muscle-derived ET(B) receptors. Competitive-binding experiments on membrane fractions by using [125I]-ET-1 and unlabeled ligands, ET-1, ET-3, sarafotoxin-6-c, and BQ-123 yielded two classes of binding sites on endothelium intact vessels from both female and male subjects. In women, the maximal binding capacities were 91+/-6 and 67+/-13 fmol/mg protein for ET(A) and ET(B) receptors, respectively; the corresponding values in men were 178+/-19 and 127+/-13 fmol/mg protein. Similar experiments performed with endothelium-denuded saphenous veins indicated the presence of both receptor subtypes on vascular smooth muscle, in contrast to our earlier report on the presence of only ET(A) receptors on vessels obtained from white Americans. Our findings demonstrate that the ratio and the density of ET receptors are different in black and white Americans. PMID- 10470989 TI - Characterization of endothelin-1 receptor subtypes in isolated human cardiomyocytes. AB - On cardiac membranes and isolated cardiomyocytes from the human heart, cell-type distribution and functional activities of endothelin-1 (ET-1) receptor subtypes were investigated by using binding methods and messenger RNA (mRNA) in situ hybridization. The ET-receptor antagonist BMS-182874 selectively and competitively inhibits ET(A) receptors both on isolated myocytes and ventricular membranes with approximately 1,300 times greater affinity for ET(A) than ET(B) subtypes. The [125I]-ET-1 specific binding revealed 42.851+/-2,546 receptors/myocyte with a prevalent proportion of ET(A)-receptor subtypes on both myocytes (84+/-2%) and ventricular membranes (66+/-3%). In situ hybridization studies revealed that mRNA for ET(A) receptors was expressed on both myocytes and nonmyocyte cells, whereas mRNA for ET(B) receptors was almost exclusively expressed on fibroblasts and endothelial cells. Specific binding of [125I]-ET-1 to both myocytes and ventricular membranes in the presence of specific ET(A) (BMS 182874) and ET(B) (BQ-788)-receptor antagonists showed a displacement of [125I] ET-1 by unlabeled ET-1, which were significantly faster from ET(B) than from ET(A). This suggests a clearance function of ventricular ET(B) receptors. PMID- 10470990 TI - Comparative study on the in vitro and in vivo activities of heparinoids derivative investigated on the animal model. AB - In this study we compared the antithrombotic and anticoagulant properties of sodium and calcium derivatives of pentosan polysulfate (Na-PPS, Ca-PPS), unfractionated heparin (UFH), and low-molecular-weight heparin (Fraxiparin). The antithrombotic effects of these agents have been investigated in an experimental thrombosis model in which rat mesenteric venules with a diameter of 20-30 microm were injured by well-defined argon laser lesions. Furthermore, the in vivo and in vitro anticoagulant activities [activated partial thromboplastin time (aPTT), Heptest] of these agents have been studied. Thrombus formation was significantly inhibited after s.c. injection of Na-PPS and Ca-PPS in doses >10 mg/kg. The duration of the antithrombotic effect lasted 8 h for Na-PPS and 12 h for Ca-PPS. After oral administration of Na-PPS, an antithrombotic effect was not observed. Oral application of Ca-PPS in doses >20 mg/kg significantly inhibited thrombus formation. Na-PPS and Ca-PPS markedly prolonged clotting time in aPTT and Heptest in concentrations ranging from 0.01 to 0.2 mg/ml rat PTT. Two h after s.c. administration of these agents in a dose of 10 mg/kg, the aPTT increased threefold and the Heptest 2.5-fold compared with controls. After oral application of 50 mg/kg Na-PPS and Ca-PPS, no effect on the coagulation test could be measured. Intravenous injection of UFH prolonged the Heptest after 1 min and the aPTT after 30 min. In ex vivo studies of aPTT and Heptest performed in rat plasma between 2 and 24 h after s.c. injection of 0.2 mg/kg Fraxiparin, no inhibition of any coagulation test was measured. The antithrombotic effect of 0.2 mg/kg Fraxiparin after s.c. injection was significant. Intravenous injection of 20 U/kg UFH significantly inhibited thrombus formation. The smallest antithrombotic effect was after i.v. injection of UFH. PMID- 10470991 TI - Acute hypotensive, natriuretic, and hormonal effects of nifedipine in salt sensitive and salt-resistant black normotensive and hypertensive subjects. AB - In a randomized double-blind study, we compared the short-term effects of nifedipine (10 mg 3x daily for 1 day) versus placebo on 24-h blood pressure, diuresis, natriuresis, urinary excretion of dopamine and metabolites, and on plasma renin activity (PRA) and plasma aldosterone levels in 18 black hypertensive (HT) patients [eight salt-resistant (HT-SR) and 10 salt-sensitive (HT-SS)], and in 20 black normotensive (NT) subjects (12 NT-SR and eight NT-SS) who were studied randomly with both a high- (HS) and a low-salt (LS) diet. In comparison to placebo, nifedipine significantly decreased 24-h mean BP in all groups either with HS or LS diets (all p<0.05). With HS, greater hypotensive effects were achieved in NT-SS (-10+/-2 mm Hg) versus NT-SR (-3+/-1 mm Hg; p<0.05) and in HT-SS (-18+/-2 mm Hg) versus HT-SR (-12+/-2 mm Hg; p<0.05). In NT SS and HT-SS, nifedipine induced greater (p<0.05) BP decrease with HS (-10+/-2 and -18+/-2 mm Hg) than with LS (-4+/-1 and -9+/-1 mm Hg, respectively), whereas in NT-SR and HT-SR, the hypotensive effect did not differ between HS and LS. Nifedipine versus placebo significantly increased natriuresis and fractional excretion of sodium in all groups only with HS (p<0.05) but not with LS diets. Only in HT-SS were the hypotensive and natriuretic effects of nifedipine significantly correlated (r = -0.77; p<0.01). Nifedipine produced a similar increase of the urinary excretion of dopamine, L-DOPA, and of DOPAC in all subjects, which did not correlate with hypotensive and natriuretic effects. Nifedipine did not modify plasma levels of renin and of aldosterone except in NT SS with HS, in whom nifedipine increased PRA levels (p <0.05). We conclude that although nifedipine reduces BP in all groups of NT and HT with LS and HS diets, the effect is greater in salt-sensitive subjects with HS. Although in HT-SS with HS, the short-term natriuretic response to nifedipine may contribute to its hypotensive effects, the diuretic-natriuretic effect of nifedipine is not necessary for the expression of its hypotensive effect. Moreover, it is unlikely that any short-term effects of nifedipine either on the renal dopaminergic system or on the secretion of aldosterone explain nifedipine short-term hypotensive and diuretic-natriuretic effects. PMID- 10470992 TI - Prolonged blockade of endothelin ET(A) receptors decreases vascular reactivity in the aorta of spontaneously hypertensive rats in vitro. AB - We investigated the effect of prolonged endothelin-1 type A (ET(A)) receptors blockade on the constrictor response to phenylephrine and the dilator response to acetylcholine (ACh) in isolated aortic rings from normotensive [Wistar-Kyoto (WKY)] rats and spontaneously hypertensive rats (SHRs). Animals were treated for 2 weeks with the ET(A)-receptor blocker LU135252 (50 mg/kg/day; n = 8). LU135252 treatment did not affect blood pressure in both strains. In isolated aortic segments, dilation to ACh and contractions to phenylephrine were decreased only in SHRs. Nitric oxide (NO) synthesis blockade (L-NAME, 0.1 mM) inhibited 90+/-11% (WKY rats) and 76+/-8% (SHRs) of ACh-induced dilation. Cyclooxygenases blockade (indomethacin, 10 microM) had no effect in both strains. Endothelium-derived hyperpolarizing factor(s) (EDHF) blockade (KCl, 20 mM) suppressed the remaining ACh-induced dilation in both strains. Treatment with LU135252 significantly decreased NO-dependent dilation, as compared with controls [70+/-8% vs. 90+/-11% (WKY rats) and 54+/-6% vs. 76+/-8% (SHRs) of total dilation; p<0.05]. On the other hand, EDHF-dependent dilation was significantly higher in the LU135252 groups [29+/-5% vs. 10+/-3% (WKY rats) and 44+/-7% vs. 19+/-4% (SHRs) of total dilation; p<0.05]. Thus prolonged ET(A)-receptor blockade decreased the responsiveness to phenylephrine and ACh in SHR aortas and changed the proportion of dilator agents in ACh-induced dilation. PMID- 10470993 TI - Characterization of endothelin receptors in the anesthetized ferret: a novel model for investigating the functional ET(B) receptor subtypes. AB - The pharmacology of endothelin (ET)-1, big ET-1, ET-3, and S6c were characterized in the anesthetized ferret to assess whether this species would provide a new and suitable nonrodent model to be used in characterization of endothelin antagonists. Unlike other species such as dog, rabbit, and rat, the ferret exhibited a dose-dependent pressor response to both ET-1 and big ET-1 with no preceding vasodilatory response. The median effective concentration (ED50) values were 0.047+/-0.009 and 0.469+/-0.003 nmol/kg for ET-1 and big ET-1, respectively. ET-3 and S6c, however, were found to elicit a transient vasodilatory response preceding the pressor response, with ED50 values of 0.23+/-0.09 and 0.18+/-0.03 nmol/kg, respectively. The rank potency of the agonists for the pressor response was found to be ET-1 > S6c > big ET-1 > ET-3. The ET(A)-specific antagonist BQ 123 was shown to block only partially the ET-1 and big ET-1 pressor response with median antagonistic dose (AD50) of 0.24+/-0.11 and 0.015+/-0.005 mg/kg, i.v., respectively, and blockade of the ET(A) receptor did not uncover an ET(B)-induced vasodilation. The dual ET(A/B) antagonist L-754,142 completely antagonized the ET 1 and big ET-1 pressor responses with AD50 values of 0.195+/-0.063 and 0.019+/ 0.006 mg/kg, respectively. The ET(B) antagonist BQ-788 blocked the depressor response of S6c entirely but was unable to antagonize the pressor response completely. BQ-123 was shown to antagonize the S6c pressor response partially, suggesting a possible interaction between the ET(A) and ET(B) receptors in the ferret. The unexpected absence of an ET-1-mediated depressor response but the presence of ET-3 and S6c vasodilation in this species supports the theory that there may be subtypes of the ET(B) receptor. These studies demonstrate that the anesthetized ferret provides a suitable model for assessing the physiological potencies of the endothelins and may provide a tool for further understanding of the diversity of the ET(B) receptor. PMID- 10470994 TI - Improvement of bradykinin endothelium-mediated vasodilation of forearm resistance circulation by quinaprilat in patients with coronary artery disease with or without left ventricular dysfunction. AB - Angiotensin-converting enzyme (ACE) inhibition potentiates bradykinin and acetylcholine endothelium-mediated vasodilation. Three groups were studied. Group I (n = 10) was the reference group; group II was composed of nine patients with coronary artery disease; and group III of seven patients with coronary artery disease and left ventricular dysfunction. Forearm blood flow was measured with plethysmography. Acetylcholine and bradykinin were administered in a random order in the brachial artery at infusion rates of 40 and 80 microg/min and 10, 30, 100 pmol/min, respectively. Then quinaprilat was infused alone at the rate of 50 microg/min and then coinfused with acetylcholine and bradykinin. Five of the reference subjects were pretreated with acetylsalicylate. Acetylcholine and bradykinin increased forearm blood flow in a dose-dependent manner in the three groups. However, the vasodilator responses to both agents were significantly lower in the two groups of patients than in the reference group. Quinaprilat significantly enhanced the vasodilator response to acetylcholine only in subjects of the reference group, whereas it enhanced the vasodilator response to each dose of bradykinin, both in subjects of the reference group and in patients. Pretreatment with aspirin did not change the vasodilator responses in any group. In healthy persons, quinaprilat had no effect on its own on forearm blood flow but enhanced the response to bradykinin and even acetylcholine. In patients with coronary disease, short-term administration of quinaprilat was able to improve the impaired response to bradykinin. The response to acetylcholine, however, could not be significantly enhanced in contrast to that in healthy subjects. PMID- 10470995 TI - The role of adenosine in insulin-induced vasodilation. AB - It was previously shown that systemic hyperinsulinemia induces vasodilation in human skeletal muscle. The mechanism mediating this vasodilation is not yet completely clarified. Based on data from animal experiments, we hypothesized that stimulation of the adenosine receptor is involved in insulin-induced vasodilation. To test this hypothesis, a 105-min hyperinsulinemic euglycemic clamp was performed in three groups of eight healthy volunteers. In group 1, placebo was infused into the left brachial artery (experimental forearm). In the second and third group, respectively, draflazine (an adenosine-uptake blocker) and theophylline (an adenosine-receptor antagonist) were administered by intrabrachial infusion. Forearm blood flow (FBF) was measured by venous-occlusion plethysmography, both at the experimental and the control forearms. The percentage decrease in flow ratio (FBF experimental arm/control arm) in the draflazine group was significantly less pronounced than that in the placebo group, whereas the percentage decrease in flow ratio was larger in the theophylline group. These results demonstrate that the insulin-induced increase in blood flow in the experimental arm was more pronounced at the site of adenosine-uptake blockade by draflazine, whereas this was reduced during adenosine-receptor antagonism by theophylline. Our observations are compatible with the hypothesis that insulin-induced vasodilation is mediated by the release of adenosine. PMID- 10470996 TI - Important role of endothelium-derived hyperpolarizing factor in shear stress- induced endothelium-dependent relaxations in the rat mesenteric artery. AB - Shear stress is one of the most important stimulators for the release of endothelium-derived relaxing factors. Although shear stress-induced release of nitric oxide (NO) has been extensively investigated, it remains to be elucidated whether endothelium-derived hyperpolarizing factor (EDHF) contributes to the endothelium-dependent relaxations to shear stress. This study was designed to address this point in the isolated rat mesenteric artery. Large mesenteric arteries (400-500 microm) and resistance mesenteric arteries (150-250 microm) of the rat were precontracted with phenylephrine (at 80 mm Hg of perfusion pressure), and the changes in vessel diameter in response to variable flow (0-300 microl/min) were continuously examined. The relative contributions of vasodilator prostaglandins, NO, and EDHF were analyzed by the inhibitory effects of indomethacin (10(-5) M), N(G)-nitro-L-arginine (L-NNA, 10(-4) M), and KCl (40 mM), respectively. The shear stress-induced relaxations were totally endothelium dependent in both-sized blood vessels, and the contribution of NO was more prominent in large arteries than in resistance arteries, whereas that of EDHF was noted in both-sized blood vessels. Tetrabutylammonium (a nonselective inhibitor of K channels) almost abolished, whereas the combination of charybdotoxin (an inhibitor of both large- and intermediate-conductance Ca2+ -activated K channels) and apamin (an inhibitor of small-conductance Ca2+ -activated K channels) significantly inhibited the EDHF-mediated component of the shear stress-induced relaxations. These results indicate that EDHF plays an important role in shear stress-induced endothelium-dependent relaxations, where K channels, especially calcium-activated K channels, appear to be involved. PMID- 10470997 TI - Presence and mechanism of direct vascular effects of amiloride in humans. AB - Besides an evident diuretic effect, amiloride has been shown to exert direct vasoactivity in various animal experiments, whereas human data on this issue are lacking. Inhibition of Na+/H+ exchange, alpha-adrenergic blockade, and sodium and calcium channel antagonism have been proposed as possible mechanisms of this action. Although the role of Na+/H+ exchange in vascular-tone modulation is not completely clear, various vasoconstrictive agents (e.g., angiotensin II) enhance its activity. We examined the direct effects of amiloride on human arterial vasculature in vivo. Forearm vasodilator responses to the infusion of placebo and amiloride (n = 10; 0.1-100 microg/min/dl) into the brachial artery were recorded by venous occlusion strain-gauge plethysmography. Reduction of forearm blood flow after local administration of noradrenaline or angiotensin II was measured before and after local amiloride administration. Amiloride increased the ratio of the infused/ noninfused forearm blood flow at the highest dosages (10, 30, and 100 microg/min/dl with 14+/-9, 17+/-14, 58+/-23% (p = 0.002, repeated-measures analysis of variance). In contrast to noradrenaline-induced vasoconstriction, the vasoconstrictor response to angiotensin II was significantly attenuated by amiloride (p = 0.02). At high concentrations, amiloride exerts direct vasodilator activity in human arterial vasculature in vivo. This effect appears not to depend on alpha-adrenergic receptor blockade, but shows interaction with angiotensin II, an activator of Na+/ H+ exchange. PMID- 10470998 TI - Protection against ventricular fibrillation by the PAF antagonist, BN-50739, involves an ischaemia-selective mechanism. AB - The platelet-activating factor (PAF) antagonist BN-50739 can suppress certain cardiac arrhythmias. PAF is released from ischaemic myocardium and may contribute to initiation of ischaemia-induced ventricular fibrillation (VF). In this study we characterised the action of BN-50739 on left regional ischaemia-induced VF and examined whether effects are mediated within the ischaemic territory, or are nonspecific. In rat isolated Langendorff perfused hearts (n = 12/group), 10 microM BN-50739 reduced the incidence of ischaemia-induced VF from 75 to 17% (p<0.05). This was accompanied by QT widening and an increase in coronary flow. Heart rate and PR interval were not affected by the drug. In separate studies, isolated rat hearts were perfused by using a dual-lumen tube that allows independent delivery of solution to the left and right coronary beds. Successful regional localisation of drug delivery was confirmed by observing, before ischaemia, a regionally selective increase in coronary flow (p<0.05), measured by using two in-line flow meters. Protection against ischaemia-induced VF (p<0.05) was achieved by pretreatment with BN-50739, delivered selectively and entrapped within the involved region, but not when the drug was delivered to the uninvolved region. In conclusion, BN-50739 protects against ischaemia-induced VF by eliciting a pharmacologic action in the involved (ischaemic) myocardium. This supports the hypothesis that BN-50739 suppresses an arrhythmogenic effect of endogenous PAF released within the ischaemic tissue. PMID- 10470999 TI - Downregulation of cardiac AT1-receptor expression and angiotensin II concentrations after long-term blockade of the renin-angiotensin system in cardiomyopathic hamsters. AB - We monitored cardiac angiotensin II concentration and AT1-receptor density after long-term blockade of the renin-angiotensin system in inbred control hamsters treated with placebo or losartan (100 mg/kg/day) and cardiomyopathic hamsters treated with placebo, low-(30 mg/kg/day), or high-dose (100 mg/kg/day) losartan or quinapril (100 mg/kg/day). All treatments were started at age 50 days. Angiotensin II-receptor density and affinity were measured by radioligand-binding assays, and ventricular angiotensin II concentration was determined by radioimmunoassay. After 125 and 275 days of treatment, both doses of losartan significantly reduced AT1-receptor density, whereas quinapril had no effect. The administration of both drugs resulted in significant reductions in ventricular angiotensin II concentration. The prolonged administration of losartan was associated with an increase in cardiac hypertrophy, suggesting that angiotensin II signaling is not directly involved or at least does not play a major role in the remodeling process observed in cardiomyopathic hamsters. PMID- 10471000 TI - Activation of soluble guanylate cyclase and potassium channels contribute to relaxations to nitric oxide in smooth muscle derived from canine femoral veins. AB - Experiments were designed to examine mechanisms of relaxations to nitric oxide (NO) in venous smooth muscle. Rings of canine femoral veins without endothelium were suspended for measurement of isometric force in organ chambers. Concentration-response curves to NO and 8-Br-cyclic guanosine monophosphate (cGMP) were obtained in veins contracted with KCl (60 mM) or prostaglandin F2alpha (PGF2alpha; 2x10(-6) M) in the absence and presence of inhibitors of soluble or particulate guanylate cyclase or K+ channel antagonists. In rings contracted with PGF2alpha, relaxations to NO were reduced significantly by inhibition of soluble but not particulate guanylate cyclase. Relaxations to NO were reduced in rings contracted with KCI. Tetraethylammonium (10(-2) M) and glibenclamide (10(-7) M) + charybdotoxin (10(-7) M) significantly reduced relaxations to NO in rings contracted with PGF2alpha. Relaxations to 8-Br-cGMP were decreased significantly only by charybdotoxin. These results suggest that relaxations to NO in canine femoral veins involve at least two intracellular processes: activation of soluble guanylate cyclase and activation of adenosine triphosphate (ATP)-sensitive and large-conductance, Ca+2-activated K+ channels. The large-conductance, Ca+2-activated K+ channels seem to be activated by cGMP dependent mechanisms. Therefore relaxations to NO in venous smooth muscle involve intracellular processes similar to those in arterial smooth muscle. PMID- 10471001 TI - Effects of prolonged exposure to ethanol in vivo on functional parameters and sensitivity to nitrendipine in the isolated rat heart. AB - This study investigated the possibility that previously reported marked upregulation of binding sites for the dihydropyridine calcium channel antagonist nitrendipine in heart tissue during the development of ethanol dependence in the rat may represent functional L-type voltage-operated calcium channels (L-VOCCs). Isolated hearts obtained from adult Sprague-Dawley rats, which received intoxicating concentrations of ethanol for 6-10 days, by inhalation, were perfused with Krebs-Henseleit solution in the Langendorff mode. Basic measurements of cardiac function were compared with hearts from control rats not exposed to ethanol vapor. In another study, concentration-response curves were constructed for nitrendipine at concentrations in the range of 10(-10)-10(-6) M for hearts obtained from control and ethanol-exposed animals. Changes in measured cardiac parameters such as R-wave amplitude, heart rate, diastolic and systolic pressure, and (+)dP/dt(max) and coronary flow were recorded. All comparisons were made between preparations set to a similar left ventricular end-diastolic pressure. Under these conditions, there were no significant differences in R-wave amplitude, but isolated hearts from ethanol-dependent rats showed significantly greater indices of myocardial contraction than did controls. These included increased systolic and developed ventricular pressure and increased (+)dP/dt(max). Coronary flow also was significantly greater in hearts from ethanol-dependent rats compared with controls. Heart rate was higher in the alcohol-exposed group, but this difference did not achieve significance. When nitrendipine was added to the perfusate at concentrations between 10(-10) and 10( 6) M, hearts from ethanol-dependent animals showed a greater sensitivity to the effects of the drug on heart rate and systolic pressure. Effects on R-wave amplitude and (+)dP/dt(max) were less clear but also suggested a greater sensitivity to nitrendipine in hearts from ethanol-exposed rats. Effects on coronary flow were small and did not differ significantly between preparations from control and ethanol-dependent rats. The results suggest that the increase in Bmax of DHP binding previously observed in hearts from ethanol-dependent animals might represent an increase in L-VOCCs, which alters physiologic function, and pharmacologic responses in the isolated heart. These changes may represent the exposure of an adaptive mechanism designed to overcome the generally depressant effects of ethanol on cardiac function in vivo. PMID- 10471002 TI - Pulmonary natriuretic peptide system during rat development. AB - Maturational changes in the rat lung natriuretic peptide system were studied postnatally in 1-, 4-, and 22-day-old rats. Lung atrial natriuretic factor (ANF) content increased significantly from day 1 to day 4 (712+/-188 vs. 1905+/-520 pg/mg protein; p<0.01) but decreased to 532+/-41 pg/mg protein, on day 22. These changes paralleled ANF messenger RNA (mRNA) detected by reverse transcribed polymerase chain reaction (RT-PCR). Rat pulmonary development also was associated with quantitative and qualitative alterations in ANF receptors. Competitive binding radioreceptor assays of lung membranes with 125I-ANF and increasing concentrations of unlabeled ANF revealed that the natriuretic peptide receptor binding sites (Bmax) progressively increased with age from 112 +/-21 fmol/mg protein at day 1 to 211+/-16 (p<0.02) and 326+/-62 fmol/mg protein (p<0.04) in 4- and 22-day-old rats, respectively. Autoradiographic studies of 125I-tyr(0)CNP binding to lung sections revealed that the levels of the natriuretic peptide receptor B (NPR-B) were undetectable. On the other hand, binding of 125I-ANF increased with age, and the higher binding at 4 days was mainly due to increased density of the clearance receptor-C (NPR-C), and at 22 days due to increased natriuretic peptide receptor-A (NPR-A). The increase in natriuretic peptide binding was confirmed at the level of synthesis, where RT-PCR revealed that NPR-A mRNA significantly increased (p<0.01) in 22-day-old rats. In conclusion, these studies demonstrate that the rat pulmonary natriuretic peptide system is altered during development. The altered synthesis of lung natriuretic peptides and their receptors may play a role in the postnatal adaptation of pulmonary circulation. PMID- 10471003 TI - Renal effects of prolonged intrarenal infusions of angiotensin II and atrial natriuretic peptide in sheep. AB - Angiotensin II (AngII) and atrial natriuretic peptide (ANP) are two hormones that have antagonistic effects on volume and pressure regulation. Plasma levels of both hormones are elevated in sheep pregnancy. However, during pregnancy, volume expansion occurs despite elevated plasma ANP, implying an overriding role of AngII. In addition to counteracting the effects of ANP on the physiological level, AngII also may act on the receptor level. Therefore this study was designed to investigate the hemodynamic and renal effects of ANP and AngII separately and to define their selective effects on the renal natriuretic peptide receptor types in the various segments of the nephron. Eight unilaterally nephrectomized nonpregnant sheep received separately for 10 days, low doses of AngII (1 ng/kg/min) and ANP (0.5 ng/kg/min) directly infused into the renal arteries to avoid systemic effects. Intrarenal AngII infusion decreased sodium excretion (UNaV) from 111+/-11 to 36 +/-8 and 45+/-6 mmol/day (p<0.05) on days 3 and 8-10, respectively. Mean arterial pressure (MAP) increased from 94 +/-6 mm Hg to a maximum of 107+/-8 mm Hg on day 5 of infusion and stabilized at 101+/-7 mm Hg on days 8-10 (p<0.05). Intrarenal ANP infusion significantly increased UNaV on day 1 from 93+/-9 to 188+/-20 mmol/day (p<0.05), followed by sodium retention on days 4-6 (average, 60+/-13 mmol/day; p<0.05). UNaV again increased above control levels on days 8-10 to an average level of 111+/-15 mmol/day. MAP decreased from 99+/-4 to 90+/-5 mm Hg (p<0.05) on days 1-3, and remained lower than control throughout the infusion period. The kidneys were collected at control nephrectomy and at the end of infusion. The natriuretic peptide receptors were characterized by competitive-binding radioreceptor assays on glomerular, outer medullary, and inner medullary membranes. AngII infusion increased the dissociation constant (Kd) of inner medullary natriuretic peptide receptors from 186 +/-11 to 267+/-22 pM (p<0.05), and ANP infusion decreased maximal binding capacity (Bmax) of inner medullary receptors from 134+/-10 to 89+/-15 fmol/mg protein (p<0.05). Glomerular and outer medullary natriuretic peptide receptors were not affected by either AngII or ANP infusion. In conclusion, AngII stimulates antinatriuresis and counteracts the hemodynamic and renal effects of ANP in part by downregulating the renal inner medullary natriuretic peptide receptors. PMID- 10471004 TI - Lidocaine inhibition of the hyperpolarization-activated current (I(f)) in sinoatrial myocytes. AB - The aim of this study was to provide information on the dose dependence and biophysical details of lidocaine blockade of the hyperpolarization-activated current (I(f)) in the sinoatrial node. Isolated rabbit sinoatrial myocytes were patch-clamped in the whole-cell configuration at 36+/-0.5 degrees C, in the presence of 1 mM Ba2+ and 2 mM Mn2+ to minimize contamination by K+ and Ca2+ currents, respectively. Lidocaine inhibited I(f) dose-dependently with a maximal inhibition of 69.5% at 75 microM and a half-maximal effect at 38.2 microM. Lidocaine reduced the conductance of fully activated I(f), without affecting the current reversal potential; the blocking effect was independent of membrane potential. Voltage dependence of I(f) activation gating was not affected by lidocaine, whose effect was independent of use and rate. Lidocaine did not modify the time course of I(f) activation. At therapeutic concentrations, lidocaine significantly inhibited I(f) by reducing fully activated channel conductance. Lack of voltage and rate dependence of effect differentiates lidocaine from most of other blockers of this current. PMID- 10471005 TI - A comparison of the antifibrillatory effects of desethylamiodarone to amiodarone in a swine model. AB - We evaluated the effect of two different doses of desethylamiodarone (DEA) and amiodarone on the ventricular fibrillation threshold (VFT). We ascertained the VFT in 24 pigs randomized to intravenous DEA, amiodarone, or vehicle. Ventricular fibrillation was induced by pacing the right ventricle by using a primary drive train at a cycle length of 270 ms for eight beats of 2-ms duration each. A secondary train of 20 pulses of 4-ms duration (100 Hz) immediately followed this over a total duration of 200 ms synchronized to the primary drive train. The intensity of the secondary train stimuli current was initially 2 mA and was increased by 2-mA increments until sustained VF with hemodynamic collapse was induced. The minimal current strength needed to induce sustained VF was defined as the VFT measured in mA. DEA (10 mg/kg) increased the VFT significantly over baseline from 13.5+/-4.9 to 23.2+/-8.8 mA (p = 0.0076). Amiodarone, 10 mg/kg, increased the VFT significantly over baseline (mean +/- SD) from 14.4+/-3.6 to 23.8+/-6.1 mA (p = 0.0016). An additional dose of amiodarone (15 mg/kg) increased the VFT to 38.5+/-15.9 mA, which is significantly greater than the VFT derived from lower-dose amiodarone (p = 0.046). We showed that DEA (10 mg/kg) has a similar antifibrillatory effect as 10 mg/kg of amiodarone. We also demonstrated a dose-dependent effect on VFT for amiodarone. PMID- 10471006 TI - The role of ATP-sensitive potassium channels in the mechanism of ischemic preconditioning. AB - We clarified the role of K(ATP) channels in the mechanism of ischemic preconditioning by using K(ATP) channel opener, nicorandil, and K(ATP) channel inhibitor, glibenclamide. Forty anesthetized dogs were divided into five groups: (a) control (C), (b) ischemic preconditioning (PC), (c) intravenous infusion of nicorandil before PC (Ni), (d) glibenclamide pretreated with PC (Gl + PC), and (e) glibenclamide pretreated with Ni (Gl + Ni). All groups were followed by 60 min ischemia and 60-min reperfusion and analyzed by the biochemical procedures. At the end of 60-min reperfusion, percentage of segment shortening in C indicated paradoxic bulging. This value was significantly recovered in PC and Ni, but it was still negative in Gl + PC and Gl + Ni. Ca2+ -adenosine triphosphatase (ATPase) activity of sarcoplasmic reticulum (SR) was significantly decreased in C. In PC and Ni, this activity was significantly maintained; however, in Gl + PC and Gl + Ni, it was similar to that in C. State III respiration of mitochondria showed similarity to the changes in SR. These results indicated that the K(ATP) channel opener enhanced the effects of ischemic preconditioning, and its blockade abolished these phenomena. We conclude that the ATP-sensitive potassium channel may play one of key roles in the mechanisms of ischemic preconditioning in the dog model. PMID- 10471007 TI - Growth hormone and hexarelin prevent endothelial vasodilator dysfunction in aortic rings of the hypophysectomized rat. AB - The endothelial vasodilation mechanism(s) has been investigated in aortic rings of hypophysectomized male rats as well as hypophysectomized rats treated for 7 days with growth hormone (GH, 400 microg/kg, s.c.) or hexarelin (80 microg/kg, s.c.). Tissue preparations from intact animals were taken as controls. The results obtained indicate that the release of 6-keto-prostaglandin F1alpha (6 keto-PGF1alpha) from aortic rings of hypophysectomized rats was markedly reduced (51%; p<0.01) as compared with that of control preparations; the peak response to cumulative concentration of endothelin-1 (ET-1, from 10(-11) to 10(-5) M) was increased 2.4-fold (p<0.01) versus controls; the relaxant activity of acetylcholine (ACh, from 10(-10) to 10(-4) M) in norepinephrine-precontracted aortic rings was reduced by 39.5+/-4.4%. Pretreatment of hypophysectomized rats with GH or hexarelin markedly antagonized the hyperresponsiveness of the aortic tissue to ET-1 and allowed a consistent recovery of both the relaxant activity of ACh and the generation of 6-keto-PGF1alpha. Collectively these findings support the concept that dysfunction of vascular endothelial cells may be induced by a defective GH function. Because a replacement regimen of GH restored the somatotropic function and increased plasma insulin-like growth factor-I (IGF-I) concentrations in the hypophysectomized rats, it is suggested that IGF-I may have protected the vascular endothelium acting as a biologic mediator of GH action. In contrast to GH, hexarelin replacement neither increased body weight nor affected the plasma concentrations of IGF-I, indicating that its beneficial action on vascular endothelium was divorced from that on somatotropic function and was likely due to activation of specific endothelial receptors. PMID- 10471008 TI - EDHF-mediated relaxation is impaired in fructose-fed rats. AB - Insulin resistance (IR) is associated with endothelial dysfunction. A defect in endothelium-dependent relaxation via outward potassium conductance has been observed in mesenteric arteries from IR rats. The purpose of this study was to assess whether this defect in endothelium-dependent relaxation was due to impaired endothelium-derived hyperpolarizing factor (EDHF) and to determine which specific potassium channel(s) are involved in relaxation. This was accomplished by using specific potassium channel inhibitors in the presence of nitric oxide synthase and cyclooxygenase inhibition. In addition, we sought to assess the function of smooth muscle cell adenosine triphosphate (ATP)-dependent potassium (K(ATP)) channels. Sprague-Dawley rats were randomized to control or IR. To determine EDHF-mediated relaxation, acetylcholine (ACh)-induced (10(-9)-10(-5) M) relaxation was measured (in vitro) in mesenteric arteries in the presence of indomethacin (10(-5) M) and N-nitro-L-arginine (L-NNA) (10(-4) M). Subsequently the combination of charybdotoxin (CTX) (0.1 microM) and apamin (0.5 microM) or glibenclamide (Glib) (10 microM) was added to the bath to inhibit KCa or K(ATP), respectively. In separate experiments, relaxation to pinacidil (10(-13)-10(-5) M), a K(ATP) activator, was assessed in vessels with intact endothelium, endothelium denuded, or with L-NNA. Maximal relaxation to ACh in the presence of L-NNA and indomethacin was 68+/-6% for control and 12+/-3% for IR (p<0.01). The addition of CTX + apamin almost abolished EDHF-mediated relaxation in control (Emax, 8+/-5% vs. 68+/-6%; p<0.01), whereas Glib had little affect. Neither CTX + apamin nor Glib had any affect on IR. Additionally, IR arteries were less sensitive to pinacidil than were controls (EC50, 1.5+/-0.9 microM vs. 5x10(-4)+/ 3x10(-4) microM, respectively; p<0.01). Endothelial removal or L-NNA pretreatment of control arteries decreased the response to pinacidil similar to IR, whereas IR vessels were unaffected. EDHF-mediated relaxation is impaired in IR arteries. In addition, the K(Ca) channel appears to be imperative for activity of EDHF in rat small mesenteric arteries. Moreover, activation of K(ATP) channels by pinacidil is impaired in IR, and this appears to be a result of endothelial dysfunction. PMID- 10471009 TI - Pulmonary artery contractility in pneumonia: role of cyclooxygenase products and nitric oxide. AB - The purpose of this study was to determine the effects of aminoguanidine (AG) and meclofenamate (MEC) on depressed contractility of small pulmonary artery (PA) rings isolated from a rat model of acute Pseudomonas pneumonia. Contractility of PA rings from lungs of control or pneumonia rats was assessed in vitro by obtaining cumulative concentration-contraction curves to potassium chloride (KCl), phenylephrine (PE), and prostaglandin F2alpha (PGF2alpha) on rings treated with or without MEC (1.0 microM), AG (100 microM), or AG + MEC. Vessels from pneumonia rats exhibited significant attenuation of the contractile responses to KCl, PE, and PGF2alpha. MEC restored the KCl and PGF2alpha contractile responses to control values, but had no effect on the attenuated PE contractile response. In contrast, AG restored the PE contractile response, and only partially affected contractile responses to KCl and PGF2alpha, MEC + AG restored the contractile responses of all three agonists. We conclude that both excess nitric oxide (NO) and cyclooxygenase products contribute to the depressed pulmonary vascular contractility observed in rats with acute pneumonia. The relative importance of NO and cyclooxygenase products in this phenomenon depends on the contractile agonist studied. PMID- 10471010 TI - Magic and methodology: when paradigms clash. PMID- 10471011 TI - Experiments, evidence, and explanations. PMID- 10471012 TI - Chinese folk medicine: the ceremony of driving away the seventy-two malignant spirits. PMID- 10471013 TI - The use of DC electrodermal potential measurements and healer's felt sense to assess the energetic nature of qi. AB - Because DC electrodermal potential measurements do not involve introducing a current into the body, it was postulated that temporal fluctuations in DC potential values on acupoints would be a useful method for assessing the subtle energetic changes of qi. DC potential measurements from acupoints and nonacupoints were therefore compared when energy healing practitioners were in an external focus state, were healing at a distance (external qi), or were self healing (internal qi). The results show statistically significant differences between measurements obtained on and off acupoints and between external focus and healing states. Subjects' report of felt sense of the flow of qi moving in their body also correlated with DC potential readings, but only those readings taken on acupoints. The results support the hypothesis that pattern information from temporal fluctuations in DC potential electrodermal measurements reflects the movement and/or amount of a physical, electrical activity that corresponds to the traditional Eastern concept of qi circulating in the body. PMID- 10471014 TI - A pilot study of electroencephalographic changes associated with ki. AB - OBJECTIVE: To measure changes in the electroencephalogram (EEG) of a ki master and ki recipients during ki emission. DESIGN: A 54-year-old, male ki master and three recipients were recorded with 21-channel digital EEG technique using the 10 20 electrode placement system. Ki recipients were three Caucasian females with no prior experience with ki practice. The ki master has practiced ki healing for more than 20 years. EEG recordings were obtained from the ki master and each recipient during baseline, direct healing ki emission, transtelephonic healing ki emission, and emission of fighting ki through the study subject to a third individual. EEG recordings were reviewed in a blinded fashion by a neurologist with no ki experience. RESULTS: No detectable EEG changes occurred during ki emission, either in the ki master or in the subjects. CONCLUSIONS: EEG may not be an appropriate objective instrument to measure ki activity, at least for moving type ki. PMID- 10471015 TI - Effects of qigong walking on diabetic patients: a pilot study. AB - OBJECTIVES: The present study was designed to evaluate the advantages of qigong walking, a mild and slow exercise that uses all the muscles of the body, in comparison with conventional walking in patients with diabetes. INTERVENTIONS: Ten inpatients with diabetes mellitus and associated complications were studied on 3 different days. Either qigong walking (30-40-minute duration) or conventional walking was performed by the patients 30 minutes after lunch on 1 of the 3 study days. Plasma glucose levels and pulse rates were measured 30 minutes after lunch and again 20 minutes after exercising; that is, 90 minutes after lunch. These data were compared to those obtained on a day with no exercise after lunch. RESULTS: Plasma glucose levels decreased during both exercises (from 228 mg/dL before to 205 mg/dL after conventional walking) and (from 223 mg/dL before to 216 mg/dL after qigong walking). In both situations the results after exercise decreased more than those in the group with no exercise (229 mg/dL; p < 0.025). The pulse rates increased after conventional walking (from 77 to 95 beats per minute; p < 0.025) and were higher than those in the group with no exercise (70 beats per minute; p < 0.01) and those after qigong walking (79 beats per minute; p < 0.05). CONCLUSIONS: Qigong walking reduced plasma glucose after lunch without inducing a large increase in the pulse rate in patients with diabetes. PMID- 10471016 TI - A study of the effect of energy healing on in vitro tumor cell proliferation. AB - This study examined the effect of energy healing on in vitro tumor cell growth using the cell culture model similar to that embraced by oncologists to assess the effect of chemotherapeutic agents. After selecting an energy healer based on his ability to influence this model, we assessed the effects of energy treatment compared to cells left at ambient temperature and to a control treatment consisting of a medical student mimicking the healer. A chi-square test comparing a medical student's and the practitioner's ability to inhibit tumor cell growth by 15% associates our practitioner with inhibition of tumor cell proliferation (p = 0.02). We also found that the magnitude of change was too close to the assay's intrinsic margin of error, thus making our quantitative data difficult to interpret. Although energy healing appears to influence several indices of growth in in vitro tumor cell proliferation, these assays are limited in their ability to define and prove the existence of this phenomenon. More sensitive biological assays are needed for further study in this field. PMID- 10471017 TI - Effects of therapeutic touch on biochemical and mood indicators in women. AB - Previous research has shown therapeutic touch (TT) to be effective in reducing anxiety and discomfort and promoting relaxation. The present investigation experimentally evaluated the effects of TT on biochemical indicators and moods in a sample of 41 healthy female volunteers. Participants were randomly assigned to either an experimental group who received TT or to a control group who did not receive TT. Pretest and posttest urine samples were collected, and personality and mood inventories were administered across three consecutive monthly sessions. Results indicated that mood disturbance in the experimental group decreased significantly over the course of the three sessions, while the control group increased in mood disturbance over time. Specifically, experimental group participants showed significant reductions in tension, confusion, and anxiety and a significant increase in vigor across sessions. Analyses of the biochemical data indicated that TT produced a significant decrease in levels of nitric oxide in the experimental group by the third TT session. The results of the present investigation have important implications for reducing symptom distress in cancer patients undergoing chemotherapy. PMID- 10471018 TI - Qigong and hypertension: a critique of research. AB - Research studies have pointed to various health-related positive effects of qigong; however, problems in much of the current qigong research methodology have limited interpretation of the potential benefits of qigong. Examples of research on qigong and hypertension are used to bring to light some of these methodological issues. In a review of 30 representative studies, various areas of concern are addressed, including: the sources of studies, lack of random assignment, selection biases, treatment effects, placebo response issues, expectancy biases, blinded outcome assessment, adherence to treatment, reliability of blood pressure measurements, regression to the mean, publication biases, and lack of consistency of measurement. One of the longest term studies, conducted by Kuang et al. (1991) and updated by Wang et al. (1993) is examined in greater detail to illustrate these issues. This study took place over 20 years and reported significant differences between a group practicing qigong and a control group in a variety of measures including a reduction of total mortality rate. The weight of evidence suggests that practicing qigong may have a positive effect on hypertension. Whether qigong alone can affect hypertension is not necessarily the most important question. Further research will be required to better assess and understand the effect of adding qigong into an integrated, multifaceted program that selectively incorporates diet, moderate aerobic exercise, relaxation training, and social and psychological dimensions. PMID- 10471019 TI - Therapeutic benefits of qigong exercises in combination with drugs. AB - This article reviews clinical studies from the Qigong Bibliographic Database, developed by the Qigong Institute, a nonprofit organization. This database was started in 1994 and holds approximately 1300 references going back to 1986, covering medical applications, scientific, and experimental studies on qigong from China, the United States, and Europe. Records in English have been compiled from International Qigong conferences and seminars, scientific journals, magazines, dissertations, MEDLINE, and other databases. The therapeutic role of qigong exercises combined with drugs is reported for three medical conditions that require drug therapy for health maintenance: hypertension, respiratory disease, and cancer. In these studies, drugs were administered to all patients who were divided into two groups, a group that practiced qigong exercises and a control group that did not. Taken together, these studies suggest that practicing qigong exercises may favorably affect many functions of the body, permit reduction of the dosage of drugs required for health maintenance, and provide greater health benefits than the use of drug therapy alone. For hypertensive patients, combining qigong practice with drug therapy for hypertensive patients resulted in reduced incidence of stroke and mortality and reduced dosage of drugs required for blood pressure maintenance. For asthma patients, the combination therapy permitted reduction in drug dosage, the need for sick leave, duration of hospitalization, and costs of therapy. For cancer patients, the combination therapy reduced the side effects of cancer therapy. Also reported is a study showing that the practice of qigong helps to rehabilitate drug addicts. The reported studies do not necessarily measure up to the strict protocols required for randomized controlled clinical trials. PMID- 10471020 TI - A PCR based DNA hybridisation capture system for the detection of human cytomegalovirus. A comparative study with other identification methods. AB - A simple, sensitive and specific colourimetric hybridisation method for the detection of HCMV DNA in clinical specimens is described. This method combines a PCR assay with a sensitive sandwich hybridisation assay. It relies on the use of a specific capture probe linked covalently to polystyrene microplates and a specific polybiotinylated detection probe. Amplified DNA fragments, sandwiched between these two probes, are detected by an enzymatic colour reaction. This PCR based colourimetric hybridisation method was compared with other known HCMV detection methods. Clinical specimens (n = 145, corresponding to 106 patients) were tested by both a nested PCR assay and this colourimetric hybridisation method; and by either the culture method or the pp65 antigenaemia test depending on the type of sample used. The results showed that the PCR-based hybridisation method has a specificity similar to tissue culture, known as the conventional gold standard method, and could be used for the examination of the clinical specimens. PMID- 10471021 TI - A comparison of in situ hybridization and immunohistochemistry for the detection of a new porcine circovirus in formalin-fixed tissues from pigs with post-weaning multisystemic wasting syndrome (PMWS). AB - Post-weaning multisystemic wasting syndrome (PMWS) is a recently identified condition affecting pigs in North America and Europe. Porcine circovirus antigen and nucleic acid have been demonstrated associated with lesions, and a new porcine circovirus designated PCV2 has been recovered from tissues of these animals. In this study, in situ hybridisation and immunohistochemical protocols were developed, optimized and compared for their relative sensitivity in detecting PCV2 antigens and nucleic acid in tissues from cases of PMWS that had been fixed for up to 6 months in formalin. For both immunohistochemistry and in situ hybridization, an increase in specific signal was observed following increased exposure to both protease XIV and proteinase K. Maximum signal and minimal loss of tissue morphology was seen after 40 min treatment with protease XIV (0.5 mg/ml). After optimisation, a comparison of these techniques on sequential sections demonstrated that both techniques successfully detected antigen or nucleic acid in all of the tissues examined. More positive cells, with increased signal intensity, were detected following immunohistochemistry. PMID- 10471022 TI - An efficient in vivo recombination cloning procedure for modifying and combining HSV-1 cosmids. AB - A helper virus-free herpes simplex virus type-1 (HSV-1) plasmid vector system developed recently may have applications in gene therapy and basic physiological studies. This system might be improved by mutating specific HSV-1 genes in the packaging system and by creating large vectors. An in vivo recombination cloning procedure is reported that supports the routine manipulation of relatively large DNAs such as the five cosmids that comprise this helper virus-free HSV-1 packaging system. In vivo recombination cloning is carried out by transforming overlapping DNA fragments into a specific RecA+ Escherichia coli, BJ5183. The cloning efficiency was improved by using a modified version of the Hanahan transformation procedure, and the background was lowered by either using vectors with different combinations of ends (5' overhangs, 3' overhangs, blunt ends) or by treating the vector with calf intestinal phosphatase. The range of usable overlap sizes is from 251 bp to 18 kb with 500 bp to 5 kb preferred. This procedure supports the routine construction and mutation of HSV-1 cosmids, by use of up to six different DNA fragments, and the construction of plasmids up to 65 kb in size. This procedure may also have applications to other vector systems and to studies on large viruses. PMID- 10471023 TI - Dobrava virus infection: serological diagnosis and cross-reactions to other hantaviruses. AB - Recent data have shown that Dobrava (DOB) hantavirus is the cause of severe haemorrhagic fever with renal syndrome (HFRS) in central and eastern Europe. To determine whether serological assays need to be based on the homologous viral antigen rather than on closely related hantavirus antigens, acute and convalescent sera from patients with HFRS collected in former Yugoslavia were examined for IgM and IgG to three hantavirus antigens; DOB, Hantaan (HTN) and Puumala (PUU). Focus reduction neutralization test was included for comparison and confirmation of the enzyme-linked immunosorbent assay (ELISA) results. Although the results showed that the cross-reactivity was high between these three antigens during the acute phase of the disease, one of 155 patients serum samples reacted only in the DOB antigen-based IgM assay. The evaluation of IgG reactivities revealed that a DOB antigen-based IgG ELISA has to be used in sero epidemiological studies; 7.1% (11/155) of the acute phase/early convalescent sera and 12.5% (2/16) of the late convalescent sera, respectively, reacted only with the homologous DOB antigen. PMID- 10471025 TI - Detection of hepatitis B virus genome in hepatocellular carcinoma tissues with PCR-in situ hybridization. AB - The detection is described of hepatitis B virus (HBV)-DNA in preserved hepatocellular carcinoma tissues, which were derived from 14 HBV-seropositive patients. Detection was by polymerase chain reaction (PCR) amplification of the target sequence, followed by specific localization of the PCR product with in situ hybridization. PISH (PCR-in situ hybridization) yielded strong positive signals in most of the tumor tissues despite very low copy numbers of chromosome integrated HBV genome, whereas no signal was detected in control samples, indicating that the signals were specific for HBV. Positive signals were sometimes detected in cirrhotic nodules surrounding the tumor regions, indicating that HBV had infected non-transformed liver cells. HBV-DNA was detected in both nucleus and cytoplasm in some specimens, possibly representing HBV at different stages of the life cycle. In one case, a gradient of viral DNA was revealed, with the highest DNA signal centered at the site of viral antigen expression. Taken together, PISH is shown to be a highly sensitive molecular detection method that is capable of detecting the presence of a low copy number viral genome in situ. PMID- 10471024 TI - Sensitive detection of potato spindle tuber and temperate fruit tree viroids by reverse transcription-polymerase chain reaction-probe capture hybridization. AB - A rapid and sensitive assay for the specific detection of plant viroids using reverse transcription-polymerase chain reaction (RT-PCR) -probe capture hybridization (RT-PCR-enzyme-linked immunosorbent assay (ELISA)) was developed. The assay was applied successfully for the detection of potato spindle tuber viroid, peach latent mosaic viroid, or apple scar skin viroid from viroid infected leaf tissue. Clarified sap extract from infected leaf tissue was treated first with GeneReleaser polymeric matrix to remove inhibitors of RT-PCR reactions. Viroid cDNA was then synthesized and amplified using viroid specific primers in RT-PCR assays and the amplified viroid cDNA (amplicon) was digoxigenin (DIG) -labelled during the amplification process. The amplicon was then detected in a colorimetric hybridization system in a microtiter plate using a biotinylated cDNA capture probe. This system combines the specificity of molecular hybridization, the ease of the colorimetric protocol, and is at least 100-fold more sensitive than gel electrophoretic analysis in detecting the amplified product. Viroid cRNA may replace viroid cDNA as the capture probe. The cRNA probe was several fold more sensitive than the cDNA probe for viroid detection. Six to seven hours are needed to complete the RT-PCR-ELISA for viroid detection from infected leaf tissue. PMID- 10471026 TI - Identification of genetic variation among St. Louis encephalitis virus isolates, using single-strand conformation polymorphism analysis. AB - A single-strand conformation polymorphism (SSCP) technique was developed for identification of genetic variation among 26 isolates of St. Louis encephalitis (SLE) virus. A 750-bp portion of the envelope gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the products analyzed by SSCP. SSCP reliably identified genetic variation among the isolates from the US, Central and South America. Closely related isolates from a smaller geographic area (Panama) were also distinguishable by SSCP. The sensitivity of this technique was demonstrated by sequencing each of the isolates used; SSCP was capable of discriminating between isolates that had as few as 1-6 nucleotide differences. These results indicate that SSCP has excellent potential as a tool to screen rapidly SLE virus isolates for genetic variation and could be incorporated into molecular epidemiology studies. PMID- 10471027 TI - Rhinovirus identification by BglI digestion of picornavirus RT-PCR amplicons. AB - Rhinoviruses are the main cause of the common cold and precipitate the majority of asthma exacerbations. RT-PCR followed by internal probe hybridisation or Southern blotting, or nested PCRs are currently the most sensitive methods for their identification. However, none of the published techniques can differentiate satisfactorily rhinoviruses from other picornaviruses. Examination of the restriction maps of sequenced rhinoviruses, revealed a highly conserved BglI restriction site (GCCnnnnnGGC), located exactly in the middle of the 380-bp amplicon generated with the OL26-OL27 primer pair, which has been used extensively in the past to identify picornaviruses. Such a site was either not present, or positioned differently in other picornaviruses of known sequence. It was, therefore, considered that digestion of rhinovirus amplicons with this enzyme would result in two equal length fragments, generating a single 190-bp band in gel electrophoresis. In contrast, either one undigested 380-bp band or a double-band pattern would appear in amplicons from other picornaviruses. To test this hypothesis, Bgl digestions of OL26-OL27 amplicons from cultured and wild type rhinoviruses, whose identity was confirmed by acid lability, as well as from echo, polio and coxsackie viruses were carried out. All rhinovirus samples were digested successfully generating single bands. Among the other picornaviruses, only 6.6% presented a single band pattern, while the rest were as predicted from the model. With a sensitivity of 100% and a specificity over 90%, the method described, which is rapid and remarkably easy to perform, can be used to distinguish rhinoviruses from other picornaviruses to a considerable extent. PMID- 10471028 TI - A novel serological technique: polymerase chain reaction enhanced immunoassay. Application to enterovirus IgM diagnosis. AB - The polymerase chain reaction (PCR) method is a sensitive, specific and rapid technique for virus detection. The principles of a PCR enhanced immunoassay (PIA) are described. The method combines solid phase serological techniques with the PCR, providing a versatile and sensitive method for antibody detection. By linking the antigenicity of virus particles with their content of nucleic acid, the method provides new possibilities for virus serology: for example, antibody specificity can be coupled to viral sequence in patients with chronic infections caused by highly variable viruses such as HIV and HCV. An application of the PIA technique is described for the detection of anti-enterovirus IgM. IgM is captured to anti-human IgM-coated microwell plates. The anti-enterovirus IgM is allowed to bind crude enterovirus antigen. Bound virus is heat denatured and the released RNA is used as a template for reverse transcription PCR (RT-PCR) amplification. Amplicons are detected by hybridisation to an affinity labelled probe in a microwell colorimetric assay. In a pilot study, 18 serum specimens from patients with enterovirus infections were examined. Using a mixture of ten crude enterovirus antigens, the frequency of IgM positivity was 6/18 (33%). Titres between 1/500 and 1/100,000 were recorded. Predominantly type-specific antibodies were detected. The results were compared with a procapsid enterovirus radioimmunoassay (RIA). After further optimisation, the PIA has the potential to be a clinically useful assay for the detection of antiviral antibodies. PMID- 10471029 TI - Comparison of two molecular techniques for the detection of avian reoviruses in formalin-fixed, paraffin-embedded chicken tissues. AB - Reverse transcription (RT) in situ polymerase chain reaction (PCR) and in situ hybridization (ISH) techniques were used to detect the sigma c-encoded gene of avian reovirus (ARV) in chicken tissue sections. The advantage of using in situ methods is to make more rapid and accurate diagnosis of ARV infections. The sensitivity of these two techniques were compared. Of the two techniques, the RT in situ PCR test was found to be more sensitive than ISH and provided the rapid, sensitive, and specific detection of ARV infections. PMID- 10471030 TI - Differentiation of closely related but biologically distinct cherry isolates of Prunus necrotic ringspot virus by polymerase chain reaction. AB - Prunus necrotic ringspot ilarvirus (PNRSV) exists as a number of biologically distinct variants which differ in host specificity, serology, and pathology. Previous nucleotide sequence alignment and phylogenetic analysis of cloned reverse transcription-polymerase chain reaction (RT-PCR) products of several biologically distinct sweet cherry isolates revealed correlations between symptom type and the nucleotide and amino acid sequences of the 3a (putative movement protein) and 3b (coat protein) open reading frames. Based upon this analysis, RT PCR assays have been developed that can identify isolates displaying different symptoms and serotypes. The incorporation of primers in a multiplex PCR protocol permits rapid detection and discrimination among the strains. The results of PCR amplification using type-specific primers that amplify a portion of the coat protein gene demonstrate that the primer-selection procedure developed for PNRSV constitutes a reliable method of viral strain discrimination in cherry for disease control and will also be useful for examining biological diversity within the PNRSV virus group. PMID- 10471031 TI - An analytical method for R5 repeated structure in varicella-zoster virus DNA by polymerase chain reaction. AB - The tandem direct reiteration, R5, in varicella-zoster virus (VZV) genome consists of 88-bp and 24-bp elements. The R5 structure was examined by the polymerase chain reaction to differentiate between various strains. A sense primer was designed at the 88-bp element and an antisense primer at the site separated from R5, in which we expected to detect multiple bands corresponding to repeating numbers of 88-bp elements. Thirty-four wild strains and a vaccine strain were analyzed using this method. The 34 wild strains showed two to four bands corresponding to numbers of 88-bp elements, in which the strain showing three bands was found to be predominant. The vaccine strain also revealed three bands. PMID- 10471032 TI - Analysis of viral glycoproteins by glycosidic digestion inside a polyacrylamide gel. AB - We adapted the method described by Cleveland et al. (1977); (Peptide mapping by limited proteolysis in sodium dodecyl sulphate and analysis by gel electrophoresis. J. Biol. Chem. 252, 1102-1106) to study the glycosidic residues linked to the viral glycoproteins of two enveloped viruses: Junin virus (JV) and rubella virus (RV). Radioiodinated glycoproteins were obtained from purified virions, isolated from SDS-polyacrylamide gels and then hydrolysed by specific glycosidases inside a second gel. N-linked oligosaccharides, mannose and galactose were found as terminal residues in the JV-G1 glycoprotein. Mannose and N-glycans of complex hybrid type were present on RV glycoproteins. PMID- 10471033 TI - Different adhesion properties of highly and poorly metastatic HT-29 colon carcinoma cells with extracellular matrix components: role of integrin expression and cytoskeletal components. AB - Integrin-mediated tumour cell adhesion to extracellular matrix (ECM) components is an important step in the development of metastatic lesions. Thus, integrin expression and integrin-mediated adhesion of colon carcinoma cells to various ECM components was examined. Poorly (HT-29P) and highly (HT-29LMM) liver-metastatic colon carcinoma cells were used to study the rates of adhesion to collagen I (C I), collagen IV (C IV), laminin (LN), fibronectin (FN), or vitronectin (VN) in a static adhesion assay (10-120 min). Cells were untreated or treated with oligopeptides (RGD, GRGDS, YIGSR, RGES), anti-integrin antibodies, or colchicine, nocodazole, cycloheximide, acrylamide or cytochalasin D (to disrupt cytoskeletal structures). Both cell lines expressed similar patterns of integrin expression (alpha2, alpha3, ,alpha6, alphav, beta1, beta4, and beta5) by immunocytochemistry and immunoprecipitation. HT-29LMM cells showed significantly higher rates of adhesion to LN (P < 0.001) and FN (P < 0.001), but significantly poorer rates of adhesion to C I (P < 0.05) and C IV (P < 0.001) than HT-29P cells, respectively, adhesion to VN was insignificant. RGD and GRGDS inhibited HT-29LMM cell adhesion to FN only. Pretreatment with anti-beta, or anti-alpha2 integrin subunits suppressed adhesion to C I and C IV, and adhesion to LN was inhibited with anti beta1 or anti-alpha6 integrin. Anti-beta1 or anti-alphav blocked adhesion to FN. Pretreatment of cells with cytochalasin D, cycloheximide or acrylamide inhibited adhesive interactions of both cell lines to the ECM components. In contrast, colchicine and nocodazole had no effect. The results demonstrate that adhesion of HT-29 cells to ECM is mediated, in part, by different integrins, depending on the substrate. Poorly and highly metastatic HT-29 cells possessed different patterns of adhesion to the various ECM substrates, but these differences were not due to different expression of integrin subunits. The results also suggested that the initial adhesion of poorly or highly metastatic HT-29 cells to ECM components requires, in part, the presence of native action and intermediate filaments, but not of microtubules. Thus the adhesion of tumour cells to ECM components may be dependent on signal transduction and assembly of microfilaments. PMID- 10471034 TI - Molecular analysis of selected cell cycle regulatory proteins during aerobic and hypoxic maintenance of human ovarian carcinoma cells. AB - We have previously reported on the development of an in vitro model system for studying the effect of hypoxia on ovarian carcinoma cell proliferation and invasion (Krtolica and Ludlow, 1996). These data indicate that the cell division cycle is reversibly arrested during the G1 phase. Here, we have continued this study to include the proliferation properties of both aerobic and hypoxic human ovarian carcinoma cells at the molecular level. The growth suppressor product of the retinoblastoma susceptibility gene, pRB, appears to be functional in these cells as determined by SV40 T-antigen binding studies. Additional G1-to-S cell cycle regulatory proteins, cyclins D and E, cyclin-dependent kinases (cdks) 4 and 2, and cdk inhibitors p27 and p18, also appear to be intact based on their apparent molecular weights and cell cycle stage-specific abundance. During hypoxia, there is a decrease in abundance of cyclins D and E, with an increase in p27 abundance. cdk4 activity towards pRB and cdk2 activity towards histone H1 are also decreased. Co-precipitation studies revealed an increased amount of p27 complexing with cyclin E-cdk2 during hypoxia than during aerobic cell growth. In addition, pRB-directed phosphatase activity was found to be greater in hypoxic than aerobic cells. Taken together, a model is suggested to explain hypoxia induced cell cycle arrest in SKA human ovarian carcinoma cells. PMID- 10471035 TI - Targeted disruption of the K-ras oncogene in an invasive colon cancer cell line down-regulates urokinase receptor expression and plasminogen-dependent proteolysis. AB - The urokinase receptor, overexpressed in invasive colon cancer, promotes tumour cell invasion. Since K-Ras is activated in many colon cancers, we determined if urokinase receptor overexpression is a consequence of this activated oncogene. Accordingly, urokinase receptor expression was compared in HCT 116 colon cancer cells containing either a mutation-activated K-Ras or disrupted for this oncogene (by homologous recombination). HCT 116 cells containing the disrupted K-Ras oncogene expressed between 50 and 85% less urokinase receptor protein compared with the parental HCT 116 cells. Reduced urokinase receptor expression in cells containing the disrupted mutated K-Ras was not due to a physical impairment of the urokinase receptor gene since phorbol ester treatment was inductive for its expression. Constitutive urokinase receptor expression in HCT 116 cells required an intact AP-1 motif in the promoter (at -184) and electrophoretic mobility shifting assays indicated less c-Jun, JunD, c-Fos and Fra-1 bound to this motif in the K-Ras-disrupted cells. Since the urokinase receptor accelerates proteolysis, laminin degradation was compared in cells containing the mutation activated and disrupted K-Ras oncogene. The latter cells displaying fewer urokinase receptors, degraded 80% less laminin. This is the first study to demonstrate a role for K-Ras as a regulator of the constitutive expression of the urokinase receptor. PMID- 10471036 TI - Acidic environment causes apoptosis by increasing caspase activity. AB - An exposure of HL-60 human promyelocytic leukaemia cells to acidic media with pH 6.2-6.6 caused an up-regulation of Bax protein expression within 2 h, which lasted for longer than 6 h. On the other hand, the apoptosis, as judged from PARP cleavage, DNA fragmentation and flow cytometric determination of cell population with sub-G1 DNA content, occurred after the cells were incubated in the acidic media for longer than 4 h. The PARP cleavage and DNA fragmentation in the cells exposed to an acidic environment could be effectively suppressed by inhibitors specific for ICE or CPP32, indicating that activation of these caspases is an essential step in acidic stress-induced apoptosis. It has been known that Bax is involved in the activation of caspases. Taken together, it appears that acidic stress first up-regulates Bax protein thereby activating caspases followed by PARP cleavage and DNA fragmentation. The observation that inhibition of either ICE or CPP32 could suppress acidic stress-induced apoptosis suggested that ICE activates pro-CPP32, which then cleaves PARP. Flow cytometric analysis indicated that acidic stress-induced apoptosis occurs mainly in G1 cells. The finding in the present study demonstrated that acidic intra-tumour environment may markedly perturb the tumour cell proliferation and tumour growth. PMID- 10471037 TI - The laminin-derived peptide YIGSR (Tyr-Ile-Gly-Ser-Arg) inhibits human pre-B leukaemic cell growth and dissemination to organs in SCID mice. AB - The YIGSR (Tyr-Ile-Gly-Ser-Arg) laminin beta1 chain sequence has an inhibitory effect on tumour growth and the metastasis of melanoma and fibrosarcoma cells. In the present study, we investigated whether the multimeric YIGSR peptide (Ac-Y16) has an antiproliferative effect and/or prevents the metastasis of human pre-B acute lymphoblastic leukaemia cells (NALM6) in severe combined immune deficient (SCID) mice. In in vitro studies, Ac-Y16 significantly inhibited leukaemic cell colony formation and the invasion of NALM6 cells in a Matrigel-based assay. The tumour growth and leukaemic infiltration in peripheral tissues were also analysed in SCID mice 9 weeks after NALM6, Matrigel and Ac-Y16 were subcutaneously co injected. The weight of the subcutaneous tumours was significantly suppressed by Ac-Y16 in a dose-dependent manner. Flow cytometry analysis showed that the leukaemic infiltration was significantly inhibited in all organs with 1.5-2.0 mg of Ac-Y16. Leukaemic infiltrations in the brain were inhibited with 0.5 mg of Ac Y16, and those in brain and bone marrow were also inhibited with 1.0 mg of Ac Y16. With Ac-S16, a control-scrambled peptide, the only significant inhibition of the leukaemic infiltration was observed in bone marrow at a much higher dose. These data suggest that the multimeric YIGSR peptide can inhibit the tumour growth and metastasis of leukaemic cells and may be useful as a potential therapeutic reagent for leukaemic infiltrations. PMID- 10471038 TI - Flavone acetic acid induces a G2/M cell cycle arrest in mammary carcinoma cells. AB - Flavone acetic acid (FAA) is a synthetic flavonoid that demonstrated extraordinary anti-tumour properties in murine models but was not effective in clinical trials. In an effort to better understand the molecular mechanisms by which FAA asserts its tumouricidal activities, we have examined the effect of FAA on the cell cycle. We observed FAA-mediated G2/M cell cycle arrest in mammary carcinoma cells at a concentration previously demonstrated to have anti-tumour effects in rodent models. The cell cycle arrest was accompanied by an increase in the P34cdc2 (cdc2) cyclin-dependent kinase activity. Morphological cytogenetic analysis demonstrated a colcemid-like effect of FAA on cytokinesis by causing accumulation of condensed C-metaphases of a sustained mitotic block. The cell cycle effect was blocked by the antioxidants ADPC and ascorbate, the superoxide scavenger Tiron, and the sphingosine kinase inhibitor L-cycloserine, but not by inhibitors of nitric oxide synthase. Based on these data, we propose that FAA may induce cell cycle arrest by stimulating the activity of acidic sphingomyelinase leading to the generation of reactive oxygen species. PMID- 10471039 TI - A novel charged trinuclear platinum complex effective against cisplatin-resistant tumours: hypersensitivity of p53-mutant human tumour xenografts. AB - Multinuclear platinum compounds were rationally designed to bind to DNA in a different manner from that of cisplatin and its mononuclear analogues. A triplatinum compound of the series (BBR 3464) was selected for preclinical development, since, in spite of its charged nature, it was very potent as cytotoxic agent and effective against cisplatin-resistant tumour cells. Anti tumour efficacy studies were performed in a panel of human tumour xenografts refractory or poorly responsive to cisplatin. The novel platinum compound exhibited efficacy in all tested tumours and an impressive efficacy (including complete tumour regressions) was displayed in two lung carcinoma models, CaLu-3 and POCS. Surprisingly, BBR 3464 showed a superior activity against p53-mutant tumours as compared to those carrying the wild-type gene. The involvement of p53 in tumour response was investigated in an osteosarcoma cell line, SAOS, which is null for p53 and is highly sensitive to BBR 3464, and in the same cells following introduction of the wild-type p53 gene. Thus the pattern of cellular response was investigated in a panel of human tumour cells with a different p53 gene status. The results showed that the transfer of functional p53 resulted in a marked (tenfold) reduction of cellular chemosensitivity to the multinuclear platinum complex but in a moderate sensitization to cisplatin. In addition, in contrast to cisplatin, the triplatinum complex was very effective as an inducer of apoptosis in a lung carcinoma cell line carrying mutant p53. The peculiar pattern of anti tumour activity of the triplatinum complex and its ability to induce p53 independent cell death may have relevant pharmacological implications, since p53, a critical protein involved in DNA repair and induction of apoptosis by conventional DNA-damaging agents, is defective in several human tumours. We suggest that the peculiar DNA binding properties of the triplatinum complex may contribute to the striking profile of anti-tumour efficacy. Taken together, the available information supports that anti-tumour activity of the novel compound is mediated by a mechanism different from that of conventional platinum complexes, and compounds of this series could represent a new class of promising anti-tumour agents. PMID- 10471040 TI - CDKN2A gene inactivation in epithelial sporadic ovarian cancer. AB - The tumour suppressor gene CDKN2A, located on chromosome 9p21, encodes the cell cycle regulatory protein p16. Inactivation of the CDKN2A gene could lead to uncontrolled cell growth. In order to determine the role of CDKN2A in the development of sporadic ovarian cancer, loss of heterozygosity at 9p21-22, homozygous deletion, mutation and methylation status of the CDKN2A gene as well as CDKN2A expression were examined in a panel of serous papillary ovarian cancer. The frequency of loss of heterozygosity (LOH) for one or more informative markers at 9p21-22 was 65% (15/23). The most common deleted region was located between interferon (IFN)-alpha and D9S171. Homozygous deletions and mutations of the CDKN2A gene were not found. There was no evidence of methylation in exon 1, but methylation in exon 2 of CDKN2A gene was found in 26% (6/23). Absence of CDKN2A gene expression was shown in 27% (6/22) at mRNA level and 21% (4/19) at protein level. These data suggest that the CDKN2A gene is involved in the tumorigenesis of ovarian cancer, but the mechanisms of CDKN2A gene inactivation in serous papillary ovarian cancer remains unclear. PMID- 10471041 TI - Laminin mediates tethering and spreading of colon cancer cells in physiological shear flow. AB - Under the physiological shear condition, cultured colon cancer cells bound to laminin (LM), but not to fibronectin or vitronectin. Most of the tethered cells did not roll, but arrested immediately and spread within 10-30 min on LM under the continuous presence of shear flow. The tethering of Colo201 was partially inhibited by monoclonal antibodies (mAbs) to alpha6 integrin and a combination of mAbs to beta1 and beta4 integrins, but not by mAb to 67KD laminin receptor. Some Colo201 cells still tethered at 4 degrees C. This suggests that alpha6beta1 and alpha6beta4 integrins participate in Colo201 tethering on LM, although other non integrin molecules play roles. In contrast, the spread of Colo201 was effectively inhibited by the mAbs to integrin alpha2, alpha6 and beta1 chains. The effect of anti-alpha2 plus anti-alpha6 mAbs was almost equal to anti-beta1, suggesting that Colo201 cells mainly use alpha2beta1 and alpha6beta1 integrins for spreading on LM. When the cells were perfused on subconfluent endothelial cells (HUVEC) cultured on LM, they did not tether on HUVEC but did on coated LM exposed at intercellular gap area. Immunohistochemistry revealed that LM abundantly existed in the cytosol of human portal and hepatic vein endothelial cells. These data suggest that LM can mediate from tethering to spreading of colon cancer cells under the blood flow and plays an essential role in haematogeneous metastasis. PMID- 10471042 TI - Tumour-specific arginine vasopressin promoter activation in small-cell lung cancer. AB - Small-cell lung cancer (SCLC) can produce numerous mitogenic neuropeptides, which are not found in normal respiratory epithelium. Arginine vasopressin is detected in up to two-thirds of SCLC tumours whereas normal physiological expression is essentially restricted to the hypothalamus. This presents the opportunity to identify elements of the gene promoter which could be exploited for SCLC-specific targeting. A series of human vasopressin 5' promoter fragments (1048 bp, 468 bp and 199 bp) were isolated and cloned upstream of a reporter gene. These were transfected into a panel of ten cell lines, including SCLC with high or low endogenous vasopressin transcription, non-SCLC and bronchial epithelium. All these fragments directed reporter gene expression in the five SCLC cell lines, but had negligible activity in the control lines. The level of reporter gene expression reflected the level of endogenous vasopressin production, with up to 4.9-fold (s.d. 0.34) higher activity than an SV40 promoter. The elements required for this strong, restricted, SCLC-specific promoter activity are contained within the 199-bp fragment. Further analysis of this region indicated involvement of E box transcription factor binding sites, although tumour-specificity was retained by a 65-bp minimal promoter fragment. These data show that a short region of the vasopressin promoter will drive strong expression in SCLC in vitro and raise the possibility of targeting gene therapy to these tumours. PMID- 10471044 TI - Treatment of malignant pericardial effusion with 32P-colloid. AB - Malignant pericardial effusion is usually treated only when signs of cardiac tamponade develop. Several methods of treatment have been reported with an overall response rate of approximately 75%. Since our initial study using intrapericardial 32P-colloid instillation as a treatment modality for pericardial effusion demonstrated a significant higher response rate, this study was conducted to further evaluate the efficacy of intrapericardial 32P-colloid in terms of response rates and duration of remissions. Intrapericardial instillation of 185-370 MBq (5-10 mCi) 32P-colloid in 36 patients with malignant pericardial effusion resulted in a complete remission rate of 94.5% (34 patients) whereas two patients did not respond to treatment due to a foudroyant formation of pericardial fluid. The median duration time was 8 months. No side-effects were observed. These results suggest that intrapericardial instillation of 32P-colloid is a simple, reliable and safe treatment strategy for patients with malignant pericardial effusions. Therefore, since further evidence is provided that 32P colloid is significantly more effective than external radiation or non radioactive sclerosing agents, this treatment modality should be considered for the management of malignant pericardial effusion. PMID- 10471043 TI - Induction of haem oxygenase-1 nitric oxide and ischaemia in experimental solid tumours and implications for tumour growth. AB - Induction of haem oxygenase-1 (HO-1) as well as nitric oxide (NO) biosynthesis during tumour growth was investigated in an experimental solid tumour model (AH136B hepatoma) in rats. An immunohistochemical study showed that the inducible isoform of NO synthase (iNOS) was localized in monocyte-derived macrophages, which infiltrated interstitial spaces of solid tumour, but not in the tumour cells. Excessive production of NO in the tumour tissue was unequivocally verified by electron spin resonance spectroscopy. Tumour growth was moderately suppressed by treatment with either Nomega-nitro-L-arginine methyl ester (L-NAME) or S methylisothiourea sulphate (SMT). In contrast, HO-1 was found only in tumour cells, not in macrophages, by in situ hybridization for HO-1 mRNA. HO-1 expression in AH136B cells in culture was strongly enhanced by an NO (NO+) donor S-nitroso-N-acetyl penicillamine. HO-1 mRNA expression in the solid tumour in vivo decreased significantly after treatment with low doses of NOS inhibitors such as L-NAME and SMT (6-20 mg kg(-1)). However, the level of HO-1 mRNA in the solid tumour treated with higher doses of NOS inhibitor was similar to that of the solid tumour without NOS inhibitor treatment. Strong induction of HO-1 was also observed in solid tumours after occlusion or embolization of the tumour feeding artery, indicating that ischaemic stress which may involve oxidative stress triggers HO-1 induction in the solid tumour. Lastly, it is of great importance that an HO inhibitor, zinc protoporphyrin IX injected intra-arterially to the solid tumour suppressed the tumour growth to a great extent. In conclusion, HO-1 expression in the solid tumour may confer resistance of tumour cells to hypoxic stress as well as to NO-mediated cytotoxicity. PMID- 10471045 TI - A population-based survey of the management of women with cancer of the cervix. AB - The aim of this study was to investigate the influence of diagnostic throughput on survival outcome for women with cancer of the cervix. We conducted a case note review of 359 women in Lancashire and Greater Manchester diagnosed with cancer of the cervix during 1990, identified from records held by the North Western Regional Cancer Registry. Univariate and multivariate survival analyses were undertaken to investigate the influence on survival of woman, disease and treatment related factors. Following adjustment for woman- and disease-related factors there was no evidence of a statistically significant association between diagnostic throughput and survival. The findings of this study do not support the need for any change in the referral patterns to gynaecologists of women with symptoms suggestive of cancer of the cervix. PMID- 10471046 TI - Induction with mitomycin C, doxorubicin, cisplatin and maintenance with weekly 5 fluorouracil, leucovorin for treatment of metastatic nasopharyngeal carcinoma: a phase II study. AB - The combination of cisplatin and 5-fluorouracil (5-FU) (PF) is the most popular regimen for treating metastatic nasopharyngeal carcinoma (NPC) but it is limited by severe stomatitis and chronic cisplatin-related toxicity. A novel approach including induction with mitomycin C, doxorubicin and cisplatin (MAP) and subsequent maintenance with weekly 5-FU and leucovorin (FL) were designed with an aim to reduce acute and chronic toxicity of PF. Thirty-two patients of NPC with measurable metastatic lesions in the liver or lung were entered into this phase II trial. Mitomycin C 8 mg m(-2), doxorubicin 40 mg m(-2) and cisplatin 60 mg m( 2) were given on day 1 every 3 weeks as initial induction. After either four courses or remission was achieved, patients received weekly dose of 5-FU 450 mg m(-2) and leucovorin 30 mg m(-2) for maintenance until disease progression. With 105 courses of MAP given, 5% were accompanied by grade 3 and 0% were accompanied by grade 4 stomatitis. The dose-limiting toxicity of MAP was myelosuppression. Forty per cent of courses had grade 3 and 13% of courses had grade 4 leukopenia. No grade 3 or 4 cisplatin-related toxicity was observed. The overall response rate was 94% (95% confidence interval (CI) 84.9-100%) with a complete response rate (CR) of 6% (95% CI: 0-15.2%) and a good partial response (PR) rate of 28% (95% CI 11.7-44.6%), which was optionally defined as observance of only equivocal lesion identifiable under imaging study. Twenty-seven cases entered weekly FL maintenance phase. The median duration of maintenance with weekly FL was 38 weeks (8-91 weeks). There was no grade 3 or 4 toxicity noted during weekly FL. The median progression-free survival and overall survival were 11.6+/-0.4 and 18.1+/ 3.6 months respectively. Six patients with a median follow-up of 19.8 months (9.6 41.0 months) were still alive and five of them had disease under control with FL. Good responders (CR and good PR) had better survival than less satisfactory responders (PR and stable disease) (P = 0.05). From Cox's multivariate regression analysis, the only significant prognostic factor for survival was good response to MAP (P = 0.042). Liver metastasis was the only significant variable in the best subset regression model that predicted good response to MAP (CR and good PR) (P = 0.027). MAP was an effective combination for metastatic NPC with minimal stomatitis and cisplatin-related toxicity but had significant myelosuppression. Weekly FL was a maintenance therapy with minimal side-effects. The response rate and overall survival of MAP-FL were better than series previously reported even when a subset of patients with poor prognosis was selected. MAP-FL's role as neoadjuvant or adjuvant therapy is worthy of further study. PMID- 10471047 TI - Somatic mutations in the p53 gene and prognosis in breast cancer: a meta analysis. AB - Many studies have investigated the association between alterations in the p53 gene and clinical outcome of breast cancer, and most investigators have reported poorer overall and disease-free survival (as indicated by a relative hazard (RH) greater than one) in breast cancer cases with somatic mutations in p53. However, different studies have produced widely differing RH estimates, ranging from no risk (RH = 1) to a relative hazard of 23, and not all of these results have been statistically significant. We have therefore reviewed all the published studies that have investigated the association between somatic mutations in the p53 gene and breast cancer prognosis and used standard techniques of meta-analysis to combine the results of these studies to produce a more precise estimate of the prognostic significance of p53 mutations. Eleven studies investigated overall survival in a total of 2319 unselected cases. The RH estimates from these ranged from 1 to 23.4 with a combined RH estimate of 2.0 (confidence interval 1.7-2.5). Three studies investigated the role of p53 in node-negative patients and in these, the combined estimate of RH was 1.7 (1.2-2.3). For three studies of node positive breast cancer the combined risk estimate was 2.6 (1.7-3.9). The inclusion of p53 mutation screening in large breast cancer clinical trials seems warranted in the light of these results. Analysis of large numbers of cases matched for stage and therapy will allow definitive clarification of the value of p53 mutational status in prognostication, and possibly choice of therapy. PMID- 10471048 TI - Primary tumour characteristics and axillary lymph node status in breast cancer. AB - This paper examines the correlation between axillary lymph node status and primary tumour characteristics in breast cancer and whether this can be used to select patients for axillary lymphadenectomy. The results are based on a retrospective analysis of 909 patients who underwent axillary dissection in our unit. Axillary lymph nodes containing metastases were found in 406 patients (44.7%), all with invasive carcinomas, but in none of the 37 carcinomas-in-situ. Nodal status was negative in all T1a tumours, but lymph node metastases were present in 16.3% and 35.7% of T1b and T1c tumours respectively. When histological grade was taken into account, positivity for grade I T1b and T1c tumours fell to 13.6% and 26.7% respectively. Lymph node metastases were found in 85% of patients with lymphovascular invasion in their tumours as compared to only 15.4% of those without and in 45.5% of oestrogen and progesterone receptor-positive tumours. When one or both hormone receptors were absent this figure was much higher. It appears that for T1a breast cancers axillary dissection is not necessary, whereas for T1b, T1c and grade I T2 tumours other histopathological parameters should be taken into consideration in deciding who should undergo axillary lymphadenectomy. PMID- 10471049 TI - Absence of mutations in the ATM gene in forty-seven cases of sporadic breast cancer. AB - Epidemiological evidence points to an increased risk of breast cancer in ataxia telangiectasia (AT) heterozygote women. Previous attempts to screen early onset or familial breast cancer patients failed to confirm an association. The issue of AT and late onset sporadic breast cancer remained unresolved. We screened 47 women who developed later onset, sporadic breast cancer for ataxia telangiectasia mutated (ATM) mutations. No mutations were found. PMID- 10471050 TI - DNA methylation analysis at distal and proximal promoter regions of the oestrogen receptor gene in breast cancers. AB - Oestrogen receptor alpha (ER-alpha) gene has two specific promoters, distal (P0) and proximal (P1), which induce almost identical transcripts in size due to different splicing. We examined the methylation at both promoter regions of the ER-alpha gene using HpaII, a methylation-sensitive restriction enzyme, prior to polymerase chain reaction (PCR) amplification. To confirm the results of PCR based methylation analysis, Southern hybridization was also performed. Twenty of 29 patients with ER-alpha-positive tumours and five of 27 with ER-alpha-negative tumours were unmethylated at the P1 promoter region of the ER-alpha gene. The incidence of methylation was highly negatively correlated with ER-alpha expression (P = 0.0002). A similarly negative correlation was observed at the P0 promoter region of the ER-alpha gene (P = 0.0154). Additionally, the tumours with the ER-alpha gene hypermethylated at both promoter regions had definitely negative ER-alpha values. It was suggested that this epigenetic change might control ER-alpha expression, and might play an important role in the loss of hormone-dependence in breast cancer. PMID- 10471051 TI - p53 protein, EGF receptor, and anti-p53 antibodies in serum from patients with occupationally derived lung cancer. AB - The oncogene product epidermal growth factor receptor (EGF-R), the tumour suppressor gene product p53 and anti-p53 antibodies are detectable in the serum of certain cancer patients. Increased levels of some of these products were reported in lung cancer patients after occupational asbestos exposure and after exposure to polycyclic aromatic hydrocarbons or vinylchloride. In the first step, this study investigated the possible diagnostic value of serum EGF-R, p53-protein and anti-p53 antibodies, measured by an enzyme-linked immunosorbent assay, in lung tumour patients. In addition to being investigated on a molecular epidemiological basis, these parameters were examined as biomarkers of carcinogenesis, especially with regard to asbestos incorporation effects or of radon-induced lung cancers. Also, a possible effect of cigarette smoking and age dependence were studied. A total of 116 male patients with lung or pleural tumours were examined. The histological classification was four small-cell cancers, six large-cell cancers, 32 adenocarcinomas, 47 squamous carcinomas, 12 mixed lung carcinomas, five diffuse malignant mesotheliomas and ten lung metastasis of extrapulmonary tumours. Twenty-two lung cancers and all mesotheliomas were related to asbestos, 22 lung cancers were related to ionizing radiation and 61 patients had cigarette smoke-related lung cancer. Besides these patients 50 male patients with non-malignant lung or pleural diseases were included; of the latter eight subjects suffered from asbestosis. Controls were 129 male subjects without any lung disease. No significantly elevated or decreased serum values for p53 protein, EGF-R, or anti-p53 antibodies as a function of histological tumour type, age, or degree and type of exposure (asbestos, smoking, ionizing radiation) could be found. The utility of p53 protein, EGF-R and anti-p53 antibodies as routine biomarkers for screening occupationally derived lung cancers is limited. PMID- 10471052 TI - Soluble CD44: quantification and molecular repartition in plasma of patients with colorectal cancer. AB - Based on the important role of CD44 in tumour progression and metastasis, we evaluated, in a prospective study, plasma-soluble CD44 (sCD44) as a serum marker in colorectal cancer. Blood plasma specimens from 89 patients with colorectal neoplasm, 22 patients with a gastrointestinal disease and 23 healthy donors were analysed for quantitation (ELISA assay) and purification of sCD44. The concentration of sCD44, indicating the concentration of all isoforms, was significantly higher in patients with colorectal cancer and intestinal disease than in normal individuals, but no significant differences were found between the two groups. We found no association between plasma levels and staging of the colorectal cancer patients according to Astler and Coller. A two-step batch purification combining ion exchange and immunoaffinity chromatography, followed by Western blot analysis, revealed a complex pattern with a major band corresponding to the standard form of CD44 and minor bands that may correspond to larger variant forms. No particular sCD44 isoform was clearly associated with anatomopathological or biological information. PMID- 10471053 TI - Expression of cyclins A and D and p21(waf1/cip1) proteins in renal cell cancer and their relation to clinicopathological variables and patient survival. AB - We have studied 118 renal cell carcinomas to analyse the expressions of cyclins A and D1 and p21(waf1/cip1), and their relationship to clinical and histopathological parameters as well as to clinical outcome. Cyclins A and D1 and cyclin-dependent kinase inhibitor p21 (waf1/cip1) were not expressed in normal renal tissue. Staining signals of cyclin D1 and p21(waf1/cip1) were always nuclear but cyclin A was also expressed in the cytoplasm of the tumour cells. The mean (range) fractions of cyclin A, cyclin D1 and p21(waf1/cip1)-positive tumour cells were 2.2% (range 0-20%), 23.3% (range 0-90%) and 6.8% (range 0-70%) respectively. The expression of cyclin A was related to venous invasion, high nuclear grade, high mitotic rate, high Ki-67 and high PCNA expressions (P < or = 0.006 for all). The expression of cyclin D1 was linked with age over 65 years, low nuclear grade and high p53 expression (P < or = 0.05 for all). An inverse correlation was present between p21(waf1/cip1) and cyclin D1 (P = 0.011). Cyclin A predicted survival in the entire study group (P = 0.0014), in T1-4/N0-2/M0 (P = 0.0007) and in T1-2/N0/M0 tumours (P = 0.0007). Cyclin A was also a powerful predictor of disease-free survival in T1-4/N0/M0 (P = 0.0027) tumours (P = 0.0007). Cyclin D1 and p21(waf1/cip1) were not significantly related to survival or disease-free survival in any of the groups. In the entire material the independent prognostic factors were the presence of distant metastases (relative risk (RR) 5.16, P < 0.001), T category (RR 2.68, P < 0.001), Ki-67 expression (RR 1.02, P = 0.026) and cyclin A expression (RR 1.12, P = 0.001). The independent predictors in T1-4/N0/M0 tumours were T-category (RR 2.67, P = 0.001) and cyclin A (RR 1.21, P < 0.001), and in T1-2/N0/M0 tumours the only significant predictor was cyclin A (RR 1.19, P = 0.0002). In renal cell carcinoma, cyclin A is a powerful and independent prognostic factor in all clinical stages of the disease, whereas cyclin D1 and p21(waf1/cip1) have no prognostic value. PMID- 10471054 TI - The molecular genetics of cervical carcinoma. AB - In the pathogenesis of cervical carcinoma there are three major components, two of them related to the role of human papillomaviruses (HPV). First, the effect of viral E6 and E7 proteins. Second, the integration of viral DNA in chromosomal regions associated with well known tumour phenotypes. Some of these viral integrations occur recurrently at specific chromosomal locations, such as 8q24 and 12q15, both harbouring HPV18 and HPV16. And third, there are other recurrent genetic alterations not linked to HPV. Recurrent losses of heterozygosity (LOH) have been detected in chromosome regions 3p14-22, 4p16, 5p15, 6p21-22, 11q23, 17p13.3 without effect on p53, 18q12-22 and 19q13, all of them suggesting the alteration of putative tumour suppressor genes not yet identified. Recurrent amplification has been mapped to 3q+ arm, with the common region in 3q24-28 in 90% of invasive carcinomas. The mutator phenotype, microsatellite instability, plays a minor role and is detected in only 7% of cervical carcinomas. The development of cervical carcinoma requires the sequential occurrence and selection of several genetic alterations. The identification of the specific genes involved, and their correlation with specific tumour properties and stages could improve the understanding and perhaps the management of cervical carcinoma. PMID- 10471055 TI - Analysis of potential markers for detection of submicroscopic lymph node metastases in breast cancer. AB - We have developed sensitive assays for cytokeratin (K) 8, 16, 19, stromelysin 3 (ST3), MUC1 and maspin mRNAs using reverse transcription polymerase chain reaction (RT-PCR) and used these to assess lymph node status in patients undergoing surgery for breast cancer. In addition the RT-PCR assays were tested against lymph nodes from non-cancer patients to determine their specificity. Despite high sensitivity RT-PCR assays for K8, K16, K19, ST3 and maspin were not found to be useful as markers of submicroscopic disease as transcripts of these genes were detected in the great majority of control lymph nodes tested. Expression of MUC1 was also not found to be useful as it was both insensitive and non-specific. The importance of assessing potential markers against an adequately sized control population is demonstrated, as failure to do so can lead to erroneous conclusions. PMID- 10471056 TI - Investigation of female survival benefit in metastatic melanoma. AB - Epidemiological studies show female survival benefit in advanced metastatic melanoma. In investigating a possible mechanism for this female survival benefit, we have previously reported that the female steroid 17beta-oestradiol significantly reduces invasion of a human melanoma cell line (A375-SM cells) and ocular melanoma cells through fibronectin. Neither cell type was found to possess oestrogen receptor-alpha. The aim of the current study was to obtain further information on the extent to which progression of cutaneous melanoma might be sex steroid sensitive by (a) examining the relationship between circulating sex steroids, sex hormone binding globulin and disease progression; (b) examining the relationship between sex steroid structure and the ability of steroids to reduce invasion of a melanoma cell line in vitro; and (c) examining the effects of sex steroids on proliferation of these cells in vitro. We report a significant reduction in circulating oestrone with disease progression in male but not female patients. Examining steroids for their ability to inhibit invasion of A375-SM cells through fibronectin in vitro, oestrogenic compounds (17beta-oestradiol and oestrone) were found to inhibit invasion; in this respect, oestrone was approximately 50 times more potent than 17beta-oestradiol; steroids lacking the benzene ring structure did not inhibit invasion, indeed dehydroepiandrosterone (DHEA) which acts as a precursor to androgenic steroids significantly enhanced invasion. Proliferation of A375-SM cells was unaffected by 17beta-oestradiol, oestrone or dihydrotestosterone when cells were cultured on plastic; in contrast, all three steroids induced modest proliferation of cells when grown on fibronectin with dihydrotestosterone the most mitogenic of the three steroids. These data are consistent with sex steroids playing a role in melanoma progression. PMID- 10471058 TI - Cytochrome P450 CYP2D6 genotypes: association with hair colour, Breslow thickness and melanocyte stimulating hormone receptor alleles in patients with malignant melanoma. AB - We previously identified associations between polymorphism in the cytochrome P450 CYP2D6 gene and outcome in several cancers. We have now examined the hypothesis that homozygosity for the mutant alleles, CYP2D6*4 and CYP2D6*3, is associated with susceptibility and outcome in malignant melanoma. Outcome was assessed by Breslow thickness. We first confirmed previous reports that these mutant alleles are associated with increased susceptibility to malignant melanoma. For example, the frequency of homozygosity for CYP2D6*4 was significantly greater (P = 0.006, chi-squared 1 d.f. = 7.4, odds ratio 2.2, 95% confidence interval 1.2, 3.9) in cases (9.1%) than in control individuals (4.3%). The frequency of homozygosity for the mutant alleles was next examined in the malignant melanoma cases grouped on the basis of characteristics associated with malignant melanoma risk. Homozygosity was significantly more common (P = 0.038) in cases with red/blonde hair than in those with brown/black hair. We found no associations between the CYP2D6 genotype and sex, skin type or eye colour. The possible association of CYP2D6 with outcome was assessed by comparing genotype frequencies in patients with tumours of Breslow thickness < 1.5 mm with those whose tumours were > or = 1.5 mm. In patients with red/blonde, but not brown or black hair, homozygosity for CYP2D6*4 was significantly associated with thicker lesions in a multivariate model (P = 0.036). We further examined the association of CYP2D6*4 homozygosity with red/blonde hair by classifying patients on the basis of homo- or heterozygosity for wild-type or val92met, asp294his or asp84glu melanocyte stimulating hormone receptor (MC1R) alleles. None of the nine patients with brown/black hair with the asp294his allele were homozygotes for CYP2D6*4. By contrast, in the patients with red/blonde hair, three of five cases with asp294his were homozygotes for the mutant CYP2D6 allele. The difference in the frequency of CYP2D6*4 homozygotes in the red/blonde cases with wild-type MC1R alleles compared with those with asp294his was significant (exact P = 0.029). No associations between val92his or asp84glu and CYP2D6 alleles were identified. PMID- 10471057 TI - Chromosomal aberrations in human hepatocellular carcinomas associated with hepatitis C virus infection detected by comparative genomic hybridization. AB - Thirty-five hepatocellular carcinomas (HCCs) associated with hepatitis C virus (HCV) were analysed by comparative genomic hybridization (CGH), to screen for changes in copy-number of DNA sequences. Chromosomal losses were noted in 1p34-36 (37%), 4q12-21 (48%), 5q13-21 (35%), 6q13-16 (23%), 8p21-23 (28%), 13q (20%), 16q (33%) and 17p13 (37%). Gains were noted in 1q (46%), 6p (20%), 8q21-24 (31%) and 17q (43%). High level gains indicative of gene amplifications were found in 7q31 (3%), 11q13 (3%), 14q12 (6%) and 17q12 (3%); amplification at 14q12 may be characteristic for HCCs. No significant difference in chromosomal aberrations was noted between carcinomas associated with HCV-infection in our study and those reported earlier in HCCs infected with hepatitis B virus (HBV), indicating that both HBV- and HCV-related carcinomas may progress through a similar cascade of molecular events. PMID- 10471059 TI - In-vivo indices of enzyme activity: the effect of renal impairment on the assessment of CYP2D6 activity. AB - Urinary drug:metabolite ratios and urinary recoveries of metabolites, have been used to assess specific enzyme activity non-invasively in vivo. These indices are potentially confounded by the effect of renal function. A recent study of the effects of renal impairment has found discrepancies between different indices used to mark CYP2D6 activity based on sparteine and dextromethorphan urinary recoveries. We have re-examined these experimental data from a theoretical viewpoint. The results suggest that the dependence of fractional urinary recovery of metabolites on renal function varies with the importance of different elimination routes. Therefore, no consistent behaviour of this index is expected when markers with different pharmacokinetics are used. However, when collecting the urine until full recovery of drug and metabolite, drug:metabolite ratios show the same degree of dependence on renal function regardless of the marker. The application of the analysis to the experimental data indicates that CYP2D6 activity is compromised in parallel with deterioration of renal function. PMID- 10471060 TI - A new variant CYP2D6 allele (CYP2D6*21) with a single base insertion in exon 5 in a Japanese population associated with a poor metabolizer phenotype. AB - Two poor metabolizer individuals of debrisoquine were identified among 215 healthy Japanese by a phenotyping test. Analysis of the CYP2D6 gene from one of two poor metabolizers, who was not homozygous for the previously described CYP2D6 variant alleles (CYP2D6*3, CYP2D6*4, CYP2D6*5 and CYP2D6*18), showed that this individual was heterozygous for a new allele, CYP2D6/C8 (CYP2D6*21). CYP2D6*21 had a single cytosine insertion at position 2661 in exon 5. This cytosine insertion generated a stop codon at the 17 bp downstream of this insertion site. A method to detect this allele was established with an allele specific-polymerase chain reaction. This method showed that another one of two poor metabolizers also possessed CYP2D6*21 allele heterozygously. In 318 healthy Japanese, five individuals carried this allele, heterozygously (0.81%, 5/636 chromosomes). Based on the present and our previous data, the poor metabolizer frequency in Japanese was estimated to be 0.39%, which accounted for approximately 45% of the individuals phenotyped as poor metabolizers by in-vivo tests. PMID- 10471061 TI - Human CYP2B6: expression, inducibility and catalytic activities. AB - Human cytochrome (CYP)2B6 cDNA was cloned and expressed in bacteria and in yeast. Its expression in Saccharomyces cerevisiae enabled us to obtain, at a high level, an active yeast-expressed CYP2B6 protein, so as to assess its role in the metabolism of ethoxyresorufin, pentoxyresorufin, benzyloxyresorufin, ethoxycoumarin, testosterone and cyclophosphamide. Kinetic analysis showed that human CYP2B6 preferentially metabolized benzyloxyresorufin and pentoxyresorufin, although other CYPs also metabolized these substrates in human liver microsomes. CYP2B6 also manifested a strong 4-hydroxycyclophosphamide activity. Its expression in Escherichia coli enabled us to produce a very specific anti-human CYP2B6 antibody. No cross reactivity of this antibody was observed with CYPs1A1, 1A2, 3A4, 3A5, 2C8, 2C9, 2C18, 2C19, 2D6 or 2E1. This antibody enabled us to study the hepatic and extrahepatic expression of CYP2B6 in man, as well as its expression and inducibility in primary cultured human hepatocytes and in different human cell lines. Immunoblot analysis revealed that the CYP2B6 protein was expressed in 43 of the 48 human liver samples tested, with levels ranging from 0.4 to 8 pmol/mg of microsomal protein with a mean of 1.7 pmol/mg protein. CYP2B was also expressed in human brain, intestine and kidney, and at a lower level in the lung. CYP2B mRNA was detected in human liver, kidney, lung, trachea and intestine. We also found that CYP2B6 is induced at protein and mRNA levels by phenobarbital (2 mM) and cyclophosphamide (1 mM), an anticancer drug known to be metabolized by CYP2B6. No expression or inducibility of CYP2B6 was observed in any of the human cell lines tested. PMID- 10471062 TI - Human nicotinamide N-methyltransferase pharmacogenetics: gene sequence analysis and promoter characterization. AB - Nicotinamide N-methyltransferase (NNMT) catalyses the N-methylation of nicotinamide and structurally related pyridines. NNMT enzymatic activity in human liver varies over a five-fold range with a bimodal frequency distribution - raising the possibility of regulation by a genetic polymorphism. We set out to characterize molecular genetic mechanisms that might be involved in the regulation of individual variation in human liver NNMT activity. After Northern blot analysis confirmed that NNMT is highly expressed in the liver, eight human hepatic biopsy samples, four each with 'low' or 'high' levels of activity, were used to perform quantitative Western blot analysis. There was a highly significant correlation (r(s) = 0.96, P < 0.0001) between NNMT activity and immunoreactive protein in these samples. We next determined that a potent promoter was located within the initial 700 bp of the 5'-flanking region of the human NNMT gene. That gene consists of 3 exons, with an initial 1240 bp intron and a second intron that is approximately 14 kb in length. We subsequently isolated DNA from 27 human liver biopsy samples with low, intermediate or high levels of NNMT activity. The three exons, all 1240 bp of intron 1 and approximately 700 bp of the 5'-flanking region of the NNMT gene were amplified from each of these samples with the polymerase chain reaction, followed by DNA sequencing to identify genetic polymorphisms that might correlate with 'NNMT phenotype'. No single nucleotide polymorphisms (SNPs) or insertion/deletion events were detected within either the exons or 5'-flanking regions of NNMT for these 27 samples. Although there were eight SNPs within intron 1, none were systematically related to level of NNMT activity. These results indicate that the exons and 5'-flanking region of the NNMT gene display little or no sequence variation. Therefore, polymorphisms within these areas of the gene are unlikely to be related to wide individual variations in the level of this enzyme activity in the human liver. PMID- 10471063 TI - Proguanil disposition and toxicity in malaria patients from Vanuatu with high frequencies of CYP2C19 mutations. AB - The increasing resistance of falciparum malaria to common antimalarial drugs has renewed interest in the compound proguanil normally metabolized to cycloguanil, a strong dihydrofolate reductase inhibitor, via the cytochrome P450 isozyme CYP2C19. The relationship between CYP2C19 genotypes and proguanil metabolism was therefore studied in 100 uncomplicated malaria patients on Malakula island in Vanuatu, where a CYP2C19-related poor metabolizer genotype status was known to be frequent. The patients (median age, 7 years) with Plasmodium falciparum or P. vivax infections, received proguanil treatment for 3 days in daily doses corresponding to adult doses of 300-500 mg. Capillary blood samples were collected on filter paper for determining both human CYP2C19 mutations by polymerase chain reaction and mutation-specific restriction enzyme digestion and blood concentrations of proguanil and its metabolites by high-performance liquid chromatography. The frequencies of the defective alleles, CYP2C19*2 and CYP2C19*3, were 0.57 and 0.25, respectively. The patients were genotyped as 68 CYP2C19-related poor metabolizers and 32 extensive metabolizers. Proguanil concentrations were higher and cycloguanil and 4-chlorophenylbiguanide concentrations were lower in poor compared to extensive metabolizers. Among the extensive metabolizers, 27 were heterozygous and five were homozygous for unmutated alleles. The tendency of an intermediate degree of proguanil metabolism in heterozygous extensive metabolizers as compared to homozygous extensive metabolizers and poor metabolizers suggests the trend towards the existence of a gene dose effect. Mild adverse events (mainly gastro-intestinal symptoms) were often reported and positively correlated with proguanil concentrations. The incidence was, however, similar in poor and extensive metabolizers. In conclusion, our data demonstrate an association between CYP2C19 mutations and poor metabolism of proguanil. PMID- 10471064 TI - Low frequency of CYP2A6 gene polymorphism as revealed by a one-step polymerase chain reaction method. AB - Human cytochrome P450 2A6 (CYP2A6) has been shown to metabolically activate carcinogens and mutagens. Genetic polymorphisms for CYP2A6 have been reported previously in different ethnic groups using a two-step polymerase chain reaction (PCR) method to identify CYP2A6*1, CYP2A6*2 and CYP2A6*3. Moreover, a new truncated allele has been recently identified in a Japanese population. We report here a one-step PCR amplification of the CYP2A6 gene from human genomic DNA and the detection of intact CYP2A6 alleles by restriction enzyme digestion. The diagnostic exon (exon 3) of the CYP2A6 gene was amplified from human genomic DNA with a primer pair. The forward primer is unique to the CYP2A6 gene, which eliminates previous problems in amplifying two highly homologous CYP2A genes, CYP2A7 and CYP2A13, in humans. The resulting PCR products (214 bp) were digested with XcmI or DdeI to detect the presence of CYP2A6*2 or CYP2A6*3 alleles, respectively. The allelic frequencies for CYP2A6*2 were 2.3% (n = 320) in the Caucasian and 0.7% (n = 71) in the Chinese populations, respectively. CYP2A6*3 allelic frequency in the Chinese population was 0.7%; while no CYP2A6*3 allele was detected in the Caucasian population. The allelic frequencies are relatively low and the reason for this discrepancy between different methods is discussed. PMID- 10471065 TI - Polymorphisms in NAT2, CYP2D6, CYP2C19 and GSTP1 and their association with prostate cancer. AB - The development of prostate cancer is dependent on heredity, androgenic influences, and exposure to environmental agents. A high intake of dietary fat is associated with an increased risk of prostate cancer, either through influence on steroid hormone profiles or through production of carcinogenic compounds that require biotransformation by enzymes. The polymorphic glutathione S-transferase (GST), N-acetyltransferase (NAT), and cytochrome P450 (CYP) enzymes are of particular interest in prostate cancer susceptibility because of their ability to metabolize both endogenous and exogenous compounds, including dietary constituents. Association between different NAT2, CYP2D6, CYP2C19 and GSTP1 genotypes and prostate cancer was studied in a Swedish and Danish case-control study comprising 850 individuals. The combined Swedish and Danish study population was analysed by polymerase chain reaction for the NAT2 alleles *4, *5A, *5B, *5C, *6 and *7, and for the CYP2D6 alleles *l, *3 and *4. The Swedish subjects were also analysed for the CYP2C19 alleles *1 and *2, and the GSTP1 alleles *A, *B and *C. No association was found between prostate cancer and polymorphisms in NAT2, CYP2D6, CYP2C19 or GSTP1. An association between CYP2D6 poor metabolism and prostate cancer was seen among smoking Danes; odds ratio 3.10 (95% confidence interval 1.07; 8.93), P = 0.03, but not among smoking Swedes; odds ratio 1.19 (95% confidence interval 0.41; 3.42), P = 0.75. Smoking is not a known risk factor for prostate cancer, and the association between CYP2D6 poor metabolism and prostate cancer in Danish smokers may have arisen by chance. PMID- 10471066 TI - UDP-glucuronosyltransferase (UGT1A1*28 and UGT1A6*2) polymorphisms in Caucasians and Asians: relationships to serum bilirubin concentrations. AB - Polymorphisms that alter UDP-glucuronosyltransferase (UGT) activities have been identified. Mutations in the promoter of the UGT1A1 gene (UGT1A1*28), resulting in 5, 7 or 8, instead of 6 thymine-adenine (TA) repeats, alter bilirubin conjugation. Two missense mutations on one allele of UGT1A6 (UGT1A6*2) result in T181A and R184S amino acid substitutions and reduced activity against phenolics, such as 4-nitrophenol, 4-hydroxycoumarin and butylated hydroxy anisole. We determined the frequency of these polymorphisms in 245 healthy men and women, aged 20-40 years and examined the relationship between TA repeat number and serum bilirubin concentrations in a subset of 24 Asians and 169 Caucasians. The frequencies of the UGT1A1*28 genotypes were 0.537, 0.348, 0.098, 0.008 and 0.008 for promoter TA repeats 6/6, 6/7, 7/7, 5/6 and 6/8, respectively. Both allele and genotype frequencies varied by race (P < 0.02), with 11% of the Caucasians and none of the Asians having the 7/7 genotype. Within both ethnic groups, serum bilirubin increased with increased numbers of UGT1A1 promoter TA repeats (P = 0.0001). However, a strong ethnic group-by-UGT1A1 genotype interaction suggests that additional ethnic differences in bilirubin metabolism contribute to observed bilirubin concentrations. Genotype frequencies for UGT1A6*2 were 0.478, 0.392, 0.029, 0.090, 0.012 for wild-type (wt)/wt, wt/T181A + R184S, wt/R184S, T181A + R184S/T181A + R184S and T181A + R184S/R184S, respectively. The co-occurrence of polymorphisms in UGT1A1 and UGT1A6 differed from that expected (P < 0.0001): individuals homozygous wild-type for UGT1A1 and UGT1A6 were observed at twice the expected frequency; individuals homozygous variant for both genes were ten-fold more frequent and individuals homozygous wild-type for one gene and homozygous variant for the other were ten-fold less frequent than expected. Overall, 8% were homozygous variant for both UGT1 polymorphisms and 43% had at least one variant allele for both UGT1A1*28 and UGT1A6*2. These highly prevalent polymorphisms, which result in modified expression and activity of UGTs, may influence susceptibility to cancers associated with altered metabolism of endogenous and xenobiotic compounds. PMID- 10471067 TI - Association of PON2 variation with birth weight in Trinidadian neonates of South Asian ancestry. AB - Variation in the PON1 and PON2 genes has been shown to be associated with coronary heart disease risk in adults of South Asian origin. In this group, low birth weight is also associated with coronary heart disease risk. We therefore hypothesized that variation in PON1 and PON2 genes may be associated with variation in birth weight. This relationship was examined in 290 consecutive Trinidadian neonates of different ethnic origins. We found that variation in PON2 was significantly associated with variation in birth weight in Trinidadian neonates of South Asian origin. Among the neonates of South Asian origin, those who were homozygous for PON2 A148/A148 had significantly lower birth weight, by approximately 200 g, compared with those with the other two genotypes (P < 0.05). For neonates of South Asian origin, PON2 A148/A148 homozygotes were significantly more prevalent in those comprising the lowest tertile for birth weight than those comprising the highest tertile (0.41 versus 0.24, P < 0.05). There were no significant associations of PON2 variation with any phenotype in other ethnic groups. We conclude that among neonates of South Asian origin, homozygosity for PON2 A148/A148, is associated with significantly lower birth weight. This suggests that genetic factors in the fetus may be important determinants of neonatal birth weight and possibly of more distal adult phenotypes, such as coronary heart disease. PMID- 10471068 TI - Polymorphic debrisoquine 4-hydroxylase activity in the rat is due to differences in CYP2D2 expression. AB - The female Dark Agouti rat is widely used as an animal model for the CYP2D6 poor metabolizer phenotype, males of other strains such as Sprague Dawley or Wistar serving as models for the extensive metabolizer phenotype. To determine the relative level of expression of CYP2D enzymes in the liver of female and male Dark Agouti, Sprague Dawley and Wistar rats, anti-peptide antibodies were raised in rabbits against short synthetic peptides representing the C-termini of the rat P450 enzymes CYP2D1, CYP2D2, CYP2D3, CYP2D4 and CYP2D5. In immunoblotting studies, it was found that the hepatic expression of CYP2D1 was greater in Dark Agouti rats than Sprague Dawley or Wistar rats. In contrast, hepatic CYP2D2 was 30-40-fold less abundant in female Dark Agouti than female Sprague Dawley or Wistar rats and six- to eightfold less abundant in male Dark Agouti than male Sprague Dawley or Wistar rats. No hepatic CYP2D3 could be detected in either sex of any of the three strains. Hepatic CYP2D4 expression was generally greater in male than female rats, and higher in Dark Agouti compared with Sprague Dawley or Wistar strains. CYP2D5 was expressed in the livers of female and male Dark Agouti rats but not in female Sprague Dawley or Wistar rats. This form was variably expressed in livers of male Sprague Dawley and Wistar rats. Hepatic debrisoquine 4-hydroxylase activity was markedly reduced in female and male Dark Agouti rats as compared to Sprague Dawley or Wistar rats and correlated (r = 0.88; P < 0.001) with the hepatic CYP2D2 content. Recombinant CYP2D2 was 18-fold more active at catalysing the 4-hydroxylation of debrisoquine than CYP2D1. Furthermore, quinine markedly inhibited CYP2D2-mediated debrisoquine and metoprolol oxidation, while quinidine, its diastereoisomer, inhibited the reactions to a lesser extent. In conclusion, these results show that impaired debrisoquine 4-hydroxylase activity in the female Dark Agouti rat is due to low levels of CYP2D2. PMID- 10471069 TI - Detailed modelling of caffeine metabolism and examination of the CYP1A2 gene: lack of a polymorphism in CYP1A2 in Caucasians. AB - The cytochrome P450 CYP1A2 is important in the metabolism of both drugs and procarcinogens such as heterocyclic amines. We aimed to clarify the existence of a phenotypic polymorphism and explore the molecular basis of such a polymorphism. Ninety-two non-smoking individuals underwent caffeine phenotyping. The distribution of the 1,7-dimethylxanthine + 1,7-dimethyluracil/caffeine (17U + 17X/137X) ratio and log-transformed data were determined. Probit plots were constructed and the distribution fitted using maximum likelihood method. The CYP1A2 gene, including upstream regulatory regions, was examined for sequence polymorphisms using the single-strand conformation polymorphism technique in 19 individuals and by complete DNA sequencing in two individuals from the extremes of the distribution. We found a similar range (1.45-18.65) and median (6.7) for the 17U + 17X/137X ratio to that found in previous studies of non-smoking Caucasians and no effect of sex. The 17U + 17X/137X ratio gave a normal distribution when log-transformed. Maximum likelihood analysis showed that the log-normal and bimodal distributions had similar deviances but the log-normal distribution was favoured because it has fewer parameters. There was no evidence for significant DNA sequence differences between fast and slow metabolizers, although some differences from published sequences including a silent polymorhpism in exon 7 which were unlikely to be of functional significance were found. We therefore conclude that CYP1A2 does not show functionally significant polymorphism but that the wide interindividual variation in activity may be due to environmental factors. PMID- 10471070 TI - Chlorzoxazone, a selective probe for phenotyping CYP2E1 in humans. AB - The ability of human cytochromes P450 other than CYP2E1 to catalyse the 6 hydroxylation of chlorzoxazone (6-OH-CHZ) was examined in vitro using human liver microsomal preparations and in vivo using chlorzoxazone as a metabolic probe. Chlorzoxazone 6-hydroxylation activity was significantly correlated with 4 nitrophenol 2-hydroxylase activity and immunodetected CYP2E1 in 14 human liver samples (r = 0.92 and 0.81, P < 0.001, respectively). Conversely, this catalytic activity was not correlated with CYP 3A or CYP1A activities. Diethyldithiocarbamate (DEDTC), a specific CYP2E1 inhibitor, reduced chlorzoxazone 6-hydroxylase activity by 92.3 +/- 7.6% (n = 14 samples) while ketoconazole, a specific CYP3A inhibitor, reduced this activity by 8.6 +/- 6.3% (n = 14). The residual activity following preincubation with DEDTC was significantly correlated with nifedipine oxidation and tamoxifen N demethylations, both specific to CYP3A (r = 0.76 and 0.68, respectively). Genetically produced pure human CYP2E1 and 3A4 hydroxylated chlorzoxazone with turnover numbers of 19.7 and 0.14 min(-1), respectively. Furthermore, cytochrome b5 stimulated chlorzoxazone 6-hydroxylation. From examination of the relative liver content of CYP2E1 and 3A, it can be asserted that CYP2E1 is the major enzyme involved in chlorzoxazone 6-hydroxylation and that the contribution of CYP3A is very minor. CYP2E1 activity was evaluated by the plasmatic metabolic ratio 6-OH-CHZ/CHZ (CHZ-MR) measured 2 h after ingestion of 500 mg CHZ. Smoker status did not influence the rate of CHZ hydroxylation. The CHZ-MR was 0.30 +/- 0.13 (mean +/- SD) n = 39 non-smokers versus 0.32 +/- 0.15, n = 75 smokers. This result suggests that CYP1A, inducible by cigarette smoking, is not significantly involved in chlorzoxazone hydroxylation. Women exhibited a slightly lower CHZ-MR than men (0.29 +/- 0.15, n = 44 versus 0.34 +/- 0.15 n = 49, respectively). Obesity increased CHZ-MR, especially in non-insulin-dependent diabetic individuals (0.45 +/- 0.21, n = 13 versus 0.30 +/- 0.15, n = 42 control individuals, P = 0.007). Furthermore, exposure of workers to volatile organics in a shoe factory decreased CHZ-MR (0.19 +/- 0.09, n = 10 Mexican workers versus 0.34 +/- 0.12, n = 16 Mexican control individuals, P = 0.001). Concomitant administration of grapefruit juice (known to be an inhibitor of CYP3A4) with chlorzoxazone did not significantly modify the CHZ metabolic ratio: 0.29 +/- 0.1 versus 0.31 +/- 0.1, for nine control individuals without and with grapefruit juice, respectively. In conclusion, all these results demonstrate that chlorzoxazone is a very selective probe for phenotyping CYP2E1 in humans. PMID- 10471071 TI - Detection of CYP2C19 alleles *1, *2 and *3 by multiplex polymerase chain reaction. PMID- 10471072 TI - A rare G2061 insertion affecting the open reading frame of CYP2D6 and responsible for the poor metabolizer phenotype. PMID- 10471073 TI - Genetic contribution to pathological gambling: possible association between a functional DNA polymorphism at the serotonin transporter gene (5-HTT) and affected men. PMID- 10471074 TI - Arylamine N-acetyltransferase 1 (NAT1) and 2 (NAT2) genes and risk of urothelial transitional cell carcinoma among Japanese. PMID- 10471075 TI - Detection of cytochrome P450 1B1 Bfr I polymorphism: genotype distribution in healthy German individuals and in patients with colorectal carcinoma. PMID- 10471076 TI - The paraxanthine:caffeine ratio in serum or in saliva as a measure of CYP1A2 activity: when should the sample be obtained? PMID- 10471077 TI - Localization of human NQO1 gene to chromosome 16q22 and NQO2-6p25 and associated polymorphisms. PMID- 10471078 TI - Memantine restores long term potentiation impaired by tonic N-methyl-D-aspartate (NMDA) receptor activation following reduction of Mg2+ in hippocampal slices. AB - This study compared the ability of memantine and (+)MK-801 to counteract deficits in the induction of long term potentiation (LTP) following reduction of Mg2+ in hippocampal slices--a model of increased synaptic noise due to tonic N-methyl-D aspartate (NMDA) receptor activation. Decreasing Mg2+ from 1 mM to 10 microM for 60 min enhanced baseline field excitatory post-synaptic potential (fEPSP) slopes (87.2 +/- 10.6% above control) and impaired LTP (-4.1 +/- 9.8% compared to pre tetanic levels). Long pre-incubations with memantine (1 microM), a concentration achieved in the CSF of dementia patients, almost fully restored the induction of LTP (to 43.4 +/- 8.4%) without changing the enhancement of baseline fEPSP slopes (84.1 +/- 11.6%). Memantine (10 microM) fully restored the induction of LTP (61.5 +/- 5.3%) and also decreased the enhancement of baseline fEPSP slopes (30.1 +/- 4.9%). In contrast, although (+)MK-801 (0.01, 0.1 and 1 microM) caused a concentration-dependent reduction in the low Mg2+ -induced enhancement of baseline fEPSP slopes, it was not able to restore the induction of LTP (3.0 +/- 9.8%, 16.3 +/- 5.7% and 4.8 +/- 6.7% respectively). These data indicate that memantine could produce symptomatological improvement in learning under conditions of tonic NMDA receptor activation such as those occurring in chronic neurodegenerative diseases whereas (+)MK-801 is likely to have only negative effects. PMID- 10471079 TI - GABA(A) receptor-mediated inspiratory termination evoked by vagal stimulation in decerebrate cats. AB - To identify the GABAergic inhibitory mechanisms involved in inspiratory termination or off-switching (IOS), the effects of a specific enhancer of GABA(A) receptors, midazolam, and an antagonist, bicuculline, on vagally evoked inspiratory inhibitions and IOS were investigated in decerebrate cats. Stimulation of vagal afferents at late inspiration provoked either reversible inspiratory inhibition or IOS, depending on the stimulus intensity. Each response occurred at a constant latency (phase 1). The reversible response was triphasic, consisting of an early (phase 2) inhibition, a brief (phase 3) excitation and a late (phase 4) inhibition in the phrenic neurogram, and early (phase 2) IPSPs, brief (phase 3) EPSPs and late (phase 4) IPSPs in bulbar inspiratory (I) neurones. With an increasing stimulus intensity, phase 4 inhibitions were increased in amplitude and duration, leading to IOS. Midazolam (0.1 mg/kg i.v.) increased more selectively phase 4 IPSPs than phase 2 IPSPs in I neurones, and decreased the threshold for evoking IOS by producing an earlier and larger phase 4 IPSPs. Bicuculline (1.0 mg/kg i.v.) had an opposite effect. These results suggest that the late inhibitory response evoked by vagal stimulation in the I neuronal pool organizes an initial phase of IOS which is mediated by GABA(A) receptors. PMID- 10471080 TI - Enhanced benzodiazepine and ethanol actions on cerebellar GABA(A) receptors mediating glutamate release in an alcohol-sensitive rat line. AB - Granule cell axon terminals of rat cerebellum possess benzodiazepine-insensitive GABA(A) receptors mediating glutamate release. We have investigated the ability of benzodiazepines, ethanol and furosemide to modulate the function of these receptors in the cerebellum of alcohol-tolerant (AT) and alcohol-nontolerant (ANT) rats. AT and ANT synaptosomes, prelabeled with [3H]D-aspartate, were superfused with GABA and various drugs during the K+ -depolarization. GABA similarly enhanced [3H]D-aspartate overflow in AT (EC50 = 1.7 microM) and ANT (EC50 = 3.9 microM) rats in a bicuculline-sensitive manner. Diazepam or zolpidem, at 0.1 microM, potentiated GABA at the GABA(A) receptor of ANT rats, but were ineffective at the AT receptor. Zolpidem acted with great potency (EC50 = 13.6 nM). Ethanol, added at 50 mM, potentiated GABA in ANT rats, but it was inactive at the GABA(A) receptor of the AT cerebellum. Furosemide significantly inhibited the effect of GABA in ANT, but not in AT synaptosomes. Our results show that one GABA(A) receptor (the receptor sited on granule cell terminals which mediates glutamate release) exhibits functional responses to diazepam and ethanol that differ between AT and ANT rats. However, the data with zolpidem and furosemide differ from previous results obtained with membranes of the granule cell layer suggesting that distinct GABA(A) receptor subtypes may exist on axon terminals versus soma/dendrites of granule cells. PMID- 10471081 TI - Abecarnil enhances recovery from diazepam tolerance. AB - Treatment with diazepam (25 mg/kg; p.o., twice-daily for 17 days) induced tolerance to the anticonvulsant effect of diazepam against bicuculline-induced convulsions in mice. Cross-tolerance was observed to the anticonvulsant action of clonazepam, imidazenil but not abecarnil. While substitution of clonazepam (12 mg/kg; p.o., twice-daily for 15 days) for diazepam did not affect tolerance to diazepam, substitution of imidazenil (17 mg/kg; p.o., twice-daily for 15 days) for diazepam significantly increased sensitivity to the anticonvulsant effect of diazepam, although tolerance was not abolished. Tolerance to diazepam progressively decreased either after suspension of diazepam administration or replacement treatment with abecarnil (20 mg/kg; p.o., twice-daily). Complete recovery of diazepam efficacy was detected after 8 and 15 days of administration of abecarnil and vehicle, respectively. Binding experiments using [3H]-flumazenil showed that Kd values did not differ among treatment groups. A significant decrease in Bmax (-42%) was observed in the cortex of diazepam-tolerant mice whether or not also treated with imidazenil and clonazepam. Conversely, chronically diazepam-treated mice, that further received abecarnil for either 8 or 15 days or vehicle for 15 days showed Bmax values similar to those of vehicle treated mice never exposed to diazepam. Results suggest that repeated abecarnil administration to diazepam-tolerant mice can facilitate re-adaptation of receptors to the diazepam-free state. It is proposed that replacement therapy with abecarnil after long-term treatment with conventional benzodiazepines (BDZs) may provide a novel approach for reducing tolerance to their anticonvulsant effects. PMID- 10471082 TI - Cannabinoid inhibition of guinea-pig intestinal peristalsis via inhibition of excitatory and activation of inhibitory neural pathways. AB - Since activation of cannabinoid CB1 receptors inhibits gastrointestinal transit in the mouse, this study analyzed the action of the cannabinoid receptor agonist methanandamide on distension-induced propulsive motility. Peristalsis in luminally perfused segments of the guinea-pig isolated ileum was elicited by a rise of the intraluminal pressure. The pressure threshold at which peristaltic contractions were triggered was used to quantify drug effects. Methanandamide (0.1-3 microM) inhibited peristalsis as deduced from a concentration-related increase in the peristaltic pressure threshold, an action that was prevented by the CB1 receptor antagonist SR141716A (1 microM) per se, which had no effect on peristalsis. The distension-induced ascending reflex contraction of the circular muscle was likewise depressed by methanandamide in a SR141716A-sensitive manner, whereas indomethacin-induced phasic contractions of the circular muscle were left unchanged by methanandamide. The anti-peristaltic action of methanandamide was inhibited by apamin (0.5 microM), attenuated by N-nitro-L-arginine methyl ester (300 microM) and left unaltered by suramin (300 microM), pyridoxal-phosphate-6 azophenyl-2',4'-disulphonic acid (150 microM) and naloxone (0.5 microM). It is concluded that methanandamide depresses intestinal peristalsis via activation of CB1 receptors on enteric neurons, which results in blockade of excitatory motor pathways and facilitation of inhibitory pathways operating via apamin-sensitive K+ channels and nitric oxide. PMID- 10471083 TI - On the effect of neonatal nitric oxide synthase inhibition in rats: a potential neurodevelopmental model of schizophrenia. AB - NADPH-d (nicotinamide-adenine dinucleotide phosphate-diaphorase) neurons are thought to migrate improperly during development in the brains of schizophrenic patients. This enzyme is a nitric oxide synthase (NOS). Nitric oxide (NO) is known to affect neurodevelopmental processes in the CNS. Therefore, we hypothesized that interference of NO generation during development may produce some aspects of schizophrenia symptomatology in a rat model. In these experiments, neonatal rats were challenged with a NOS inhibitor (L-nitroarginine 1-100 mg/kg s.c.) daily on post-natal days 3-5. L-Nitroarginine (L-NoArg) treated male rats developed a hypersensitivity to amphetamine in adulthood versus vehicle treated controls, whereas female rats did not. However, L-NoArg treated female rats developed a hypersensitivity to phencyclidine (PCP) at juvenile and adult ages versus vehicle treated controls, whereas male animals did not. L-NoArg treated male rats also had deficits in pre-pulse inhibition of startle whereas adult female rats did not. The results are discussed in terms of a new neurodevelopmental model of schizophrenia and male/female differences inherent in this disease. PMID- 10471084 TI - Neuroprotective effects of DETA-NONOate, a nitric oxide donor, on hydrogen peroxide-induced neurotoxicity in cortical neurones. AB - Nitric oxide (NO) has been proposed to exert neuroprotective actions against oxidative damage acting directly as an antioxidant; in addition, it has also been suggested that NO might be cytoprotective by increasing cyclic GMP concentrations via activation of soluble guanylate cyclase. In this context, we have previously shown that cyclic GMP elevations confer cytoprotection against the neurotoxicity induced by SIN-1 in the presence of superoxide dismutase, conditions in which cell death seems to be a consequence of hydrogen peroxide (H2O2) formation. We have now found that H2O2 (20-100 microM) causes neurotoxicity in 1-week-old rat cortical neurones and that this effect is inhibited by the NO donor DETA-NONOate (1-10 microM). We have also found that 1H-[1,2,4]oxadiazolo[4,3,-alpha]quinoxalin 1-one (ODQ), a selective inhibitor of soluble guanylate cyclase, reverses the effect induced by DETA-NONOate, and that this action of ODQ is mimicked by 8-(4 chlorophenylthio)guanosine-3',5'-monophosphorothioate (Rp-8-pCPT-cGMPS), an inhibitor of cyclic GMP-dependent protein kinase, suggesting that the pathway affording protection involves activation of this kinase by cyclic GMP elevations. Simultaneously, ODQ inhibits the elevation of cyclic GMP concentrations induced by DETA-NONOate (1-3 microM) in cortical cells. Finally, we have also shown that the cyclic GMP mimetic, 8-bromoguanosine 3':5'-cyclic monophosphate (8-Br-cyclic GMP) inhibits the neurotoxicity induced by H2O2 (30-40 microM). Taken together, these data demonstrate that NO-induced cyclic GMP elevations confer cytoprotection against H2O2-induced neuronal cell death. PMID- 10471085 TI - The neuropeptide FF analogue, 1DME, enhances in vivo met-enkephalin release from the rat spinal cord. AB - Behavioural studies have suggested that endogenous opioids mediate the antinociceptive action of neuropeptide FF (FLFQPQRF-NH2) at the spinal level in the rat. This hypothesis was directly assessed by investigating the effects of a NPFF analogue, 1DMe ([D-Tyr1,(NMe)Phe3]NPFF), on the spinal outflow of met enkephalin-like material (MELM) in halothane-anaesthetised rats. Intrathecal infusion (0.1 ml/min) of 1DMe (0.1 microM-0.1 mM, for 45 min) produced a concentration-dependent increase in spinal MELM outflow which persisted for at least 90 min at the highest concentration tested. Intrathecal coadministration of the micro-opioid receptor antagonist CTOP (1 microM) did not significantly affect the spinal MELM overflow due to 0.1 mM 1DMe. In contrast, both naltrindole and nor-binaltorphimine, at concentrations (10 microM) that allow the selective blockade of alpha- and kappa-opioid receptors, respectively, significantly reduced the stimulatory effect of 1DMe on spinal MELM outflow. These data provide the first direct demonstration that met-enkephalin (among other opioid peptides) can mediate the antinociceptive action of NPFF at the spinal level in rats. In addition, they suggest that reciprocal excitatory interactions between opioids and opioid-modulatory factors (such as NPFF) participate in the physiological control of nociception. PMID- 10471086 TI - The non-steroidal anti-inflammatory drug tepoxalin inhibits interleukin-6 and alpha1-anti-chymotrypsin synthesis in astrocytes by preventing degradation of IkappaB-alpha. AB - Tepoxalin is a structurally and functionally novel non-steroidal anti inflammatory drug (NSAID) with potent anti-inflammatory and analgesic properties. Apart from its inhibitory effect on cyclooxygenase activity, tepoxalin is able to inhibit production of cytokines in peripheral cells outside the CNS. No data, however, are available concerning the effects of this drug in the CNS. Since cytokines such as interleukin-1 (IL-1) or interleukin-6 (IL-6) as well as acute phase proteins such as alpha1-anti-chymotrypsin (ACT) participate in the etiopathology of Alzheimer's disease (AD), we were interested whether tepoxalin is able to inhibit the synthesis of these immunomodulators in primary rat microglia and astrocytes as well as in the human astrocytoma cell line U373 MG. We found that tepoxalin markedly inhibits IL-1beta-induced IL-6 and ACT synthesis in astrocytes and the synthesis of IL-1beta and IL-6 in lipopolysaccharide (LPS) stimulated microglial cells. Electrophoretic mobility shift and reporter gene assays revealed that tepoxalin exerts its inhibitory effect through the inhibition of nuclear factor kappaB (NF-kappaB), a transcription factor involved in the induction of IL-1, IL-6 and ACT gene expression. We show that inhibition of NF-kappaB activation by tepoxalin is mediated by preventing IkappaB-alpha degradation. Based on this inhibitory effect of tepoxalin on cytokine and ACT synthesis and the documented therapeutic efficacy of NSAIDs in AD, we conclude that tepoxalin may be of therapeutic benefit for the treatment of AD patients and should therefore be tested in clinical trials. PMID- 10471087 TI - Neuroprotective effects of modulators of P2 receptors in primary culture of CNS neurones. AB - In previous studies (Volonte and Merlo, 1996. J. Neurosci. Res. 45, 183-193) basilen blue was shown to be a P2 receptor antagonist which abrogated glutamate mediated cytotoxicity in cerebellar neurones in primary culture. Our work has now been extended to evaluate the neuroprotective action of the compound in additional neuronal systems, as well as in a different paradigm of cell death. We show that basilen blue prevents L-glutamate-mediated neurotoxicity in rat cerebellar (90-100% inhibition), cortical (60-70%) and hippocampal (50%) neurones. Similarly, glutamate-dependent progressive darkening of cell bodies, loss of phase-brightness and rapid cellular swelling are inhibited. Basilen blue is significantly less toxic and more effective at blocking L-glutamate toxicity in mixed cortical/glial cultures, compared to its structural analogue cibacron blue. Moreover, its neuroprotective effect is correlated with the time of incubation with granule neurones. Other purinoceptor ligands, including 2,2' pyridylisatogen, but not pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid 4 sodium, are also effective in preventing glutamate toxicity. Furthermore, basilen blue prevents serum deprivation- and low potassium-induced apoptotic cell death in cerebellar granule neurones. In summary, our data extend and reinforce the possibility of a potential therapeutic use of P2 receptor modulators as neuroprotective agents for the central nervous system. PMID- 10471088 TI - Neuroprotective effects of early vs. late administration of dantrolene in experimental status epilepticus. AB - We examined the effects of dantrolene, an inhibitor of intracellular calcium release, on hippocampal neuronal damage associated with 140 min of limbic status epilepticus in the rat. Dantrolene (10 mg/kg i.p.) was administered after either 30 min or 140 min of status epilepticus. Early administration was associated with a significant reduction in the amount of neuronal injury in all hippocampal subregions, while late administration was associated with less neuronal injury in area CA3 only. These findings suggest that a substantial portion of seizure induced hippocampal injury is associated with release of calcium from intracellular stores, and that early administration of dantrolene may be a useful adjunct to anticonvulsant treatment of status epilepticus. PMID- 10471089 TI - Carbamazepine inhibits L-type Ca2+ channels in cultured rat hippocampal neurons stimulated with glutamate receptor agonists. AB - In order to better understand the mechanism(s) of action of carbamazepine (CBZ), we studied its effects on the increase in [Ca2+]i and [Na+]i stimulated by glutamate ionotropic receptor agonists, in cultured rat hippocampal neurons, as followed by indo- or SBFI fluorescence, respectively. CBZ inhibited the increase in [Ca2+]i stimulated either by glutamate, kainate, alpha-amino-3-hydroxy-5 methyl-isoxazole-4-propionate (AMPA), or N-methyl-D-aspartate (NMDA), in a concentration-dependent manner. In order to discriminate the effects of CBZ on the activation of glutamate receptors from possible effects on Ca2+ channels, we determined the inhibitory effects of Ca2+ channel blockers on [Ca2+]i changes in the absence or in the presence of CBZ. The presence of 1 microM nitrendipine, 0.5 microM omega-conotoxin GVIA (omega-CgTx GVIA), or of both blockers, inhibited the kainate-stimulated increase in [Ca2+]i by 51.6, 32.9 or 68.7%, respectively. In the presence of both 100 microM CBZ and nitrendipine, the inhibition was similar (54.1%) to that obtained with nitrendipine alone, but in the presence of both CBZ and omega-CgTx GVIA, the inhibition was greater (54%) than that caused by omega CgTx GVIA alone. However, CBZ did not inhibit the increase in [Na+]i stimulated by the glutamate receptor agonists, but inhibited the increase in [Na+]i due to veratridine. Tetrodotoxin, or MK-801, did not inhibit the influx of Na+ stimulated by kainate, indicating that Na+ influx occurs mainly through the glutamate ionotropic non-NMDA receptors. Moreover, LY 303070, a specific AMPA receptor antagonist, inhibited the [Na+]i response to kainate or AMPA by about 70 or 80%, respectively, suggesting that AMPA receptors are mainly involved. Taken together, the results suggest that CBZ inhibits L-type Ca2+ channels and Na+ channels, but does not inhibit activation of glutamate ionotropic receptors. PMID- 10471090 TI - Effects of nimodipine on extracellular dopamine levels in the rat nucleus accumbens in ethanol withdrawal. AB - Withdrawal from chronic ethanol intoxication is associated with a reduction of dopamine neurotransmission. However, the mechanisms of dopamine depletion, a putative neurochemical correlate of the dysphoric symptomatology, are not yet understood. To assess the role of L-type calcium channels in the inhibition of the dopaminergic system in the withdrawal state, the effects of the dihydropyridine calcium channel antagonist nimodipine on the extracellular levels of dopamine were studied in the nucleus accumbens shell of awake rats 10 h after withdrawal from chronic ethanol intoxication. In control, chronic sucrose withdrawn rats, nimodipine did not change extracellular dopamine levels. However, in ethanol-withdrawn rats nimodipine (5 or 10 mg/kg s.c.) increased extracellular dopamine to 136 +/- 16 and 305 +/- 19% of pre-administration values, respectively, the latter dose elevating levels above those of controls. The elevations of extracellular DA by nimodipine (10 mg/kg) were associated with a significant reduction (-17%) of the overall behavioural score of the withdrawal symptomatology, as evaluated for 11 behavioural items. Significant reductions of the score for convulsions (-47%) and, to a lesser extent, for catatonia (-30%) and tremors (-15%) contributed to the overall effect. It is suggested that overactivity of L-type calcium channels is involved in the mechanisms of dopamine depletion as well as in certain behavioural/neurological signs associated with ethanol withdrawal. By restoring depleted dopamine levels, dihydropyridines might ameliorate the dysphoric symptoms of ethanol abstinence. PMID- 10471091 TI - Interaction between L-DOPA and 3-O-methyl-L-DOPA for transport in immortalised rat capillary cerebral endothelial cells. AB - The present study aimed to determine the kinetics of L-3,4-dihydroxyphenylalanine (L-DOPA) uptake in an immortalised cell line of rat capillary cerebral endothelial cells (clones RBE 4 and RBE 4B), to define the type of interaction with 3-O-methyl-L-DOPA (3-OM-L-DOPA), sensitivity to 2-aminobicyclo(2,2,1) heptane-2-carboxylic acid (BHC), N-(methylamino)-isobutyric acid (MeAIB) and sodium. Non-linear analysis of the saturation curves for L-DOPA and 3-OM-L-DOPA revealed in RBE 4 cells Km values (in microM) of 72 (53, 91) and 40 (25, 57) and in RBE 4B cells Km values (in microM) of 60 (46, 74) and 44 (13, 75), respectively. IC50 values for 3-OM-L-DOPA (RBE 4, 642 [542, 759] microM; RBE 4B, 482 [475, 489] microM) obtained in the presence of a nearly saturating (250 microM) concentration of L-DOPA were greater than the corresponding Ki values (RBE 4, 143 [121, 170] microM; RBE 4B, 93 [92, 95] microM) obtained in the presence of a nearly saturating (250 microM) concentration of 3-OM-L-DOPA; this is compatible with a competitive type of interaction between L-DOPA and 3-OM-L DOPA. Uptake of both L-DOPA and 3-OM-L-DOPA in RBE 4 and RBE 4B cells was sensitive to BHC with similar IC50 values. MeAIB (up to 2.5 mM) was found not to interfere with the uptake of both L-DOPA and 3-OM-L-DOPA. Uptake of (250 microM) L-DOPA and 3-OM-L-DOPA in the absence of sodium in the incubation medium was similar to that observed in the presence of increasing concentrations of sodium (20-140 mM). Homogenates of both cell lines were endowed with considerable COMT activity. Incubation of RBE 4 and RBE 4B cells with L-DOPA (25 microM) in the presence of a methyl donor (S-adenosyl-L-methionine) resulted in the formation of 3-OM-L-DOPA; this was abolished by 1 microM tolcapone. The fractional outflow of intracellular L-DOPA through the luminal and abluminal cell side was not affected by the presence of intracellular 3-OM-L-DOPA. The fractional outflow of exogenous 3-OM-L-DOPA applied from the luminal cell border was similar to that observed for 3-OM-L-DOPA with origin in L-DOPA. It is concluded that RBE 4 and RBE 4B cells are endowed with the L-type amino acid transporter through which L-DOPA and 3-OM L-DOPA can be taken up, and 3-OM-L-DOPA behaves as a competitive inhibitor for the uptake of L-DOPA. This, however, only occurs for luminal cell inward movement but not for abluminal cell outward movement of the substrates. PMID- 10471092 TI - Pseudoephedrine, a sympathomimetic agent, induces Fos-like immunoreactivity in rat nucleus accumbens and striatum. AB - The pharmacological properties of the ephedrine derivative pseudoephedrine were investigated at the nuclear level. Following intraperitoneal injection of Sprague Dawley rats with pseudoephedrine, Fos induction was measured in various brain areas by Western blots and immunocytochemistry. Pseudoephedrine induced Fos-like immunoreactivity in the nucleus accumbens and striatum in a time and concentration-dependent manner with maximal effect at 60 mg/kg 2 h after injection. Immunocytochemical studies confirmed that the majority of the signal was detectable in the nucleus accumbens and striatum. Pre-injection with the D1 dopamine receptor antagonist SCH23390 partially and completely blocked pseudoephedrine-induced Fos-like immunoreactivity in the striatum and nucleus accumbens, respectively, suggesting that the action of pseudoephedrine is mediated via dopamine release and results in the activation of D1 dopamine receptors. With the exception of the higher doses required, the actions of pseudoephedrine were similar to those previously described for the psychostimulant amphetamine. PMID- 10471093 TI - Dopamine D2 receptor knock-out mice are insensitive to the hypolocomotor and hypothermic effects of dopamine D2/D3 receptor agonists. AB - The dopamine (DA) D2-like family of receptors is comprised of three subtypes, the D2, D3, and D4 receptors. It has been suggested that the potency of DA receptor agonists to produce hypothermia and hypolocomotion in rodents correlates more strongly with the in vitro affinity for, or potency (mitogenesis test) at the D3 than at the D2 subtype. However, it has recently been reported that when tested in DA D3 receptor knock-out mice, several DA D2/D3 receptor agonists (7-OH-DPAT, PD 128907 and quinelorane) induced levels of hypothermia and decreases of locomotor activity similar to those obtained in control (wild-type) mice. These results do not argue in favour of an implication of DA D3 receptors in these in vivo effects. In order to investigate whether the DA D2 receptor is the subtype that mediates hypothermia and hypolocomotion produced by DA D2/D3 receptor agonists, we tested the effects of ip administration of the DA D2/D3 receptor agonists 7-OH-DPAT and PD 128907, on core temperature and locomotor activity in DA D2 receptor knock-out mice (homozygotes: D2(-/-) and heterozygotes: D2(+/-)), and in wild-type (D2(+/+)) mice. 7-OH-DPAT (0.1-3 mg/kg) and PD 128907 (1-10 mg/kg) induced hypothermia and decreased locomotion in D2(+/+) mice, but had no effects in D2(-/-) mice; the magnitude of the hypothermic and locomotor-reducing effects of these two agonists in D2(+/+) mutants was approximately half that of D2(+/+) mice. During the first 10 min in the activity chambers, the level of spontaneous locomotor activity of D2(-/-) individuals was almost 50% below that of D2(+/+) mice; basal locomotor activity of D2(+/-) mice was between that of D2( /-) and D2(+/+) individuals. Neither type of mutant showed spontaneous catalepsy or deficits in forelimb muscle strength (grip-strength test). These results show that the presence of DA D2 receptors is necessary for the expression of the locomotor- and core temperature-decreasing effects of DA D2/D3 receptor agonists such as 7-OH-DPAT and PD 128907. PMID- 10471094 TI - Behavioural and neurochemical effects of cholinergic and dopaminergic agonists administered into the accumbal core and shell in rats. AB - The first goal of this study was to investigate whether turning behaviour elicited by unilateral injections of the cholinergic agonist carbachol into the shell of the nucleus accumbens differs from that elicited by similar injections into the core of this nucleus, and to compare the behavioural effects with the known effects of such injections of the mixture of the dopamine D1 and D2 receptor agonists SKF 38393 (5 microg) and quinpirole (10 microg). The second goal was to investigate whether these injections of carbachol produce neurochemical alterations in the ventrolateral striatum that differ from similar injections of the mixture of the dopamine D1 and D2 receptor agonists into these brain regions. Injections of carbachol into the shell produced predominantly (a) contralateral circling marked by normal stepping and running in wide circles during the initial 50 min and (b) postural asymmetry during the following 75 min; similar injections into the core produced (a) contralateral pivoting, namely pathological head-to-tail turning marked by abnormal hindlimb stepping during the initial 50 min and (b) postural asymmetry during the next 75 min. The postural asymmetry seen after the carbachol injections was closely associated with the drug-induced increase in the dopamine release measured by microdialysis in the ipsilateral striatum. Injections of the mixture of dopamine agonists into the shell, but not core, also produced pivoting. These shell injections increased the dopamine release in the ipsilateral striatum, and decreased it in the contralateral striatum. The relative increase in the ipsilateral striatum was closely associated with the drug-induced pivoting. The data show that stimulation of cholinergic and dopaminergic receptors in the shell and core elicit effects that vary according to the subregion of the nucleus accumbens. It is concluded that the accumbens-specific, cholinergic effects are mediated via substrates that differ from those involved in the shell-specific, dopaminergic effects. PMID- 10471095 TI - Enhancement of 5-HT1B and 5-HT1D receptor antagonist effects on extracellular 5 HT levels in the guinea-pig brain following concurrent 5-HT1A or 5-HT re-uptake site blockade. AB - The effects of selective serotonin re-uptake inhibitor (SSRI), paroxetine, and 5 HT1A, 5-HT1B and 5-HT1B/1D receptor antagonists on in vivo extracellular 5-HT levels in the guinea-pig frontal cortex and dorsal hippocampus were investigated using the technique of microdialysis. The aim of the study was to further investigate the autoreceptor roles of the 5-HT1A, 5-HT1B and 5-HT1D receptors in the median vs dorsal raphe nuclei. In the frontal cortex, 5-HT1A (WAY 100635, 1 mg/kg i.p.) or 5-HT1B (SB-224289, 4 mg/kg i.p.) receptor antagonists had no effect on extracellular levels of 5-HT, whilst the mixed 5-HT1B/1D receptor antagonist (GR 127935, 0.3 mg/kg i.p) produced a significant decrease in extracellular 5-HT levels. Paroxetine (10 microM) significantly increased extracellular 5-HT levels when perfused locally into the cortex. Administration of SB-224289, followed 120 min later by WAY 100635, had no effect on extracellular 5-HT levels. In contrast, sequential administration of either WAY 100635 and GR 127935, or SB-224289 and paroxetine significantly increased extracellular 5-HT levels. In the dorsal hippocampus, whilst 5-HT1A receptor antagonism elicited by administration of WAY 100635 had no effect, both 5-HT1B and mixed 5-HT1B/1D receptor blockade significantly increased extracellular 5-HT levels. Administration of SB-224289 followed 120 min later with WAY 100635, or WAY 100635 followed 30 min later with GR 127935, potentiated the effect of the three compounds alone, significantly increasing extracellular 5-HT levels. These data demonstrate that to simultaneously increase extracellular 5-HT in both frontal cortex and dorsal hippocampus of the guinea-pig brain concurrent 5-HTA1A, 5-HT1B and 5-HT1D receptor blockade is required. Whereas in the dorsal hippocampus, 5-HT1B receptor blockade is sufficient to elicit an increase in extracellular 5-HT levels. PMID- 10471096 TI - The effects of the peptide-coupling agent, EEDQ, on 5-HT2A receptor binding and function in rat frontal cortex. AB - This ex vivo study in rat frontal cortex determined the influence of 5-HT receptor agonists and antagonists on EEDQ-induced depletion of 5-HT2A binding sites and reduction in their functional coupling to phospholipid hydrolysis. Twenty-four hours after EEDQ (6 mg/kg) administration a marked reduction (66%) of cortical 5-HT2A binding sites with no change in binding affinity was observed. The 5HT2A antagonists ritanserin (1 mg/kg), ketanserin (1 and 5 mg/kg), metergoline (3 mg/kg) or the 5HT2A agonist, DOI (3 and 10 mg/kg) also significantly reduced (by 15-44%) these binding sites 24 h after injection. Thirty minute pretreatment with ritanserin, ketanserin, metergoline or DOI (at the doses above) afforded 49-65% protection against the loss of 5-HT2A binding sites induced by EEDQ (6 mg/kg). DOI (10 mg/kg) pretreatment (-24 h) decreased by 26% the accumulation of [3H]inositol phosphates (IPs) evoked by 5-HT (100 microM), but did not affect that produced by DOI (100 microM). Ketanserin (5 mg/kg, -24 h) decreased 5-HT- and DOI-induced IP formation by 65% and 53%, respectively. The EEDQ (6 mg/kg, -24 h)-evoked reductions (-50%) of 5-HT- and DOI induced IP formation were not altered by DOI (10 mg/kg) or ketanserin (5 mg/kg) given 30 min before EEDQ. G-protein-stimulated IP accumulation was unaffected by EEDQ (6 mg/kg). Overall, EEDQ reduces 5-HT2A binding sites and function in rat frontal cortex, whereas its effects on binding were attenuated by various 5-HT receptor antagonists and agonists, its effects on function was unaltered by these drugs. PMID- 10471097 TI - Surveillance of sexually transmitted infections: fit for purpose? PMID- 10471098 TI - Palliative care and AIDS: 2--Gastrointestinal symptoms. AB - With the use of more intensive antiretroviral therapies (highly-active antiretroviral therapy, HAART) particularly in first world countries, reductions in the mortality and morbidity of HIV infection are being seen. However, though the prevalence of symptoms may change, symptom control does continue to be a problem for many people with HIV, particularly as their disease progresses. This is the second of 2 CME articles about palliative care and HIV infection. The first gave a background to palliative care, and covered symptom control of pain. This article gives suggestions for the treatment of common gastrointestinal symptoms in HIV infection; nausea and vomiting, cachexia and anorexia and chronic diarrhoea. PMID- 10471099 TI - Universal prophylaxis for Chlamydia trachomatis and anaerobic vaginosis in women attending for suction termination of pregnancy: an audit of short-term health gains. AB - A previous study of infection and morbidity in 400 women attending for termination of pregnancy (TOP) had shown that 32 (8%) harboured cervical Chlamydia trachomatis and 112 (28%) had anaerobic (bacterial) vaginosis (AV). Fifty-three per cent of the women with preoperative C. trachomatis had AV. Thirty of the 32 women with chlamydial infection were followed up and 19 (63%) of these developed post-abortion upper genital tract infection, 7 of whom needed re admission. In view of the high morbidity in women with chlamydial infection attending for TOP, anti-bacterial prophylaxis with metronidazole suppositories and oral oxytetracycline was introduced for women attending for suction termination of pregnancy (STOP). An audit of the clinical and financial benefits and/or losses was carried out. The audit of 1951 consecutive patients attending for STOP revealed that 132 (6.8%) had chlamydial infection with equivocal results reported in a further 2 patients. One hundred and eight of the 134 women responded to recall. Full genital tract infection screening was carried out in 105 of the 108 recalled patients of whom 5 had repeat positive cervical swabs for C. trachomatis, one had Trichomonas vaginalis, 24 had candidiasis and 17 had anaerobic vaginosis, none had gonorrhoea. Thirteen (12%) of the 108 women had pelvic infection as previously defined, none of whom required re-admission. At least pound sterling 20,000 has been saved each year in our Trust following the introduction of pre-abortion chlamydial screening and universal antichlamydial and anti-anaerobe prophylaxis. The introduction of universal prophylaxis against C. trachomatis and AV has profoundly reduced morbidity in patients attending for TOP and has also resulted in substantial financial savings. PMID- 10471100 TI - Long-term immunotherapy in HIV infection, combined with short-term antiretroviral treatment. AB - Forty asymptomatic HIV-infected individuals with CD4+ lymphocyte levels above 400x10(6)/l were immunized over 5 years with recombinant envelope glycoprotein gp160 (rgp160). After 5 years there was a trend towards more non-progressors in the immunized group as compared to the matched controls. Since immunizations could activate HIV, the first 6 immunizations were followed by 2 weeks of zidovudine or placebo, double-blind. The viral load did not change during the first 6 months and was not different from that of the matched controls after 5 years. The best effect on CD4+ lymphocyte development was seen in individuals with a high viral load randomized to rgp160+zidovudine and in individuals with a low viral load randomized to rgp160+placebo. We conclude that rgp160 is safe and results in temporarily improved CD4+ development. Concomitant antiviral treatment might be of benefit, especially in patients with a more advanced disease and can today be given with more effective combinations. PMID- 10471101 TI - Seroprevalence of herpes simplex virus type 2 antibodies in an STD clinic in Paris. AB - Our objective was to evaluate the seroprevalence of herpes simplex virus (HSV)-2 and HSV-1 in a population of men and women attending the STD clinic of Hopital St Louis (Paris, France). Four hundred and eighty-seven patients (264 men and 223 women) were tested for HSV-2 and HSV-1 antibodies by specific enzyme immunoassay (EIA) (Smithkline-Beecham Biologicals). Univariate and multivariate analyses were carried out for correlations with clinical, socio-epidemiological and behavioural data. HSV-2 seroprevalence was 55% (44.7% in men, 67.3% in women). HSV-1 seroprevalence was 93% (94.7% in men, 91% in women). The predictive factors of HSV-2 seropositivity being female (OR: 3.37), age (OR: 1.04), country of origin (Central Africa OR: 3.52, North Africa OR: 1.36), history of genital herpes (OR: 10.97), hepatitis B virus (HBV) markers (OR: 1.92) and hepatitis C virus (HCV) markers (OR: 3.96). The only protective factor was HSV-1 seropositivity (OR: 0.25). The predictive factors of HSV-1 seropositivity were only the country of origin (Central Africa OR: 2.95, North Africa OR: 1.83) and the absence of genital herpes (OR: 11.01). Only 23 (8.6%) HSV-2 seropositive patients had a history of genital herpes. This study underlines the very high HSV-2 seroprevalence of patients with STDs, only a few of whom have a history of genital herpes. Detection and counselling is urgently needed for these patients. PMID- 10471102 TI - Diagnostic tests and specimens used to screen for Chlamydia trachomatis in genitourinary medicine clinics in the United Kingdom. AB - This questionnaire study looked at the diagnostics tests and specimens used to screen for Chlamydia trachomatis in UK genitourinary medicine (GUM) clinics. Replies were received from 70% (185/265) of clinics. Half used only one site to screen women. One-third took anal swabs from patients who had anal sex and 10% took oropharyngeal swabs from patients who had oral sex. Immunoassays were used to screen men for chlamydia in 86% of the clinics and women in 88%. Only 60% of male and 62% of female immunoassays were supplemented by a second test. Six per cent of clinics used molecular technique (MT) to screen men and 4% to screen women and 4% were trying to acquire it. Culture was not available to 58% of clinics. MT was not available to 81%, 89% of which was due to non provision locally and/or cost. Only 7% of clinicians thought that using MT for screening was unnecessary. There were significant differences in the availability of the technique between large academic and small clinics. A national review of GUM strategies to screen for C. trachomatis with adequate funding is urgently needed. PMID- 10471103 TI - Post-traumatic stress disorder in female survivors of rape attending a genitourinary medicine clinic: a pilot study. AB - This study describes psychological symptomatology including post-traumatic stress disorder (PTSD) in 19 women attending a specialist sexual assault service within a genitourinary medicine (GUM) clinic. Women were interviewed within one year post-rape (mean = 12 weeks) using standardized questionnaires for PTSD and other psychological symptomatology. Seventeen (89.5%) of 19 women met full criteria for a diagnosis of PTSD. Anxiety predominated amongst other psychological symptomatology. Suicidal ideation was reported by 8 women and one made a suicide attempt following the rape. Although it is acknowledged this is a small, select sample, the high level of psychological trauma found suggests that genitourinary medicine clinics providing for sexual assault require access to mental health professionals. PMID- 10471104 TI - Contact tracing for gonorrhoea in homosexual and heterosexual men. AB - We aimed to determine whether the success of partner notification for gonorrhoea in men was affected by sexual orientation. Analysis of standard clinic and health adviser records of all male patients found to be infected with gonorrhoea between October 1992 and September 1993 were carried out. Of the 278 cases of gonorrhoea in men, 9% (25) were acquired through homosexual intercourse and 91% (253) by heterosexual contact. Fifteen per cent (24) of Caucasians were homosexual but only 0.9% (1) of Afro-Caribbeans were. The mean number of contacts was 1.36 for homosexuals and 1.38 for heterosexuals. Contact information was given by 55% of heterosexuals and 48% of homosexuals. The proportion of acknowledged contacts attending was 38% for homosexuals and 56% for heterosexuals (P = 0.054). Fifty two per cent of homosexuals and 59% of heterosexuals had at least one contact attend. Data analysis on Caucasians only showed Caucasian gay men had a higher mean number of contacts (1.38) than Caucasian heterosexuals (1.28). Caucasian homosexuals had a lower proportion of contacts attending (40% vs. 77%) (P = 0.05), 54% of homosexual men and 60% of heterosexual men had at least one contact attending (P = 0.74). There is a trend for partner notification to be less successful in homosexual men when all ethnic groups are considered together. In Caucasian men with gonorrhoea, homosexuals have a greater number of partners than heterosexuals and have a lower proportion of total contacts attending but there is no difference in the proportion having at least one contact attending. Data on sexual orientation and ethnicity should be reported in studies assessing efficacy of contact tracing. PMID- 10471105 TI - Nursing in genitourinary medicine: 10 years on from the Monks Report. The membership of GUNA. Genito-Urinary Nurses Association. AB - Various reports and surveys since 1988 have highlighted differences in gradings and diversity of roles and responsibilities of nurses in genitourinary medicine (GUM). In addition there was no method of defining how many nurses work in the speciality. Therefore the Genito-Urinary Nurses Association (GUNA) undertook a survey last year to establish how many nurses, of what grade, undertaking what roles are working in GUM clinics. Over half the clinics in the UK responded, from full-time busy London clinics to small part-time rural clinics. The results suggest approximately 2000 nurses working in GUM nationally, with a wide diversity of grades, roles and responsibilities still apparent 10 years after the Monks Report. Further analysis of the data obtained by the survey is given in the report. PMID- 10471106 TI - HIV/STD: the women to blame? Knowledge and attitudes among STD clinic attendees in the second decade of HIV/AIDS. AB - We aimed to determine the knowledge and attitudes towards HIV/STDs among women attending an STD clinic by interviewing 520 randomly selected women. Nearly all had heard of HIV/AIDS/STDs, with posters, pamphlets and the radio being the main source of their information. The years of schooling was the only predictive factor of knowing a preventive measure of HIV. Two-thirds thought they were at risk of contracting HIV from their regular partner. Knowledge of the sexual habits of their male partners was low with 260 (50%) of the women distrusting their partner. Only 52 (10%) of respondents admitted to sex in exchange for gifts or money. In the event of a positive HIV test result, the perceived partner response would be to blame the woman for introducing the infection into the relationship. After a positive HIV test result, only 3.5% would resort to using condoms while another 3.7% would try to pass on the disease to other people. The quality of their knowledge of the transmission of HIV was low in spite of the fact that most respondents have heard of HIV/AIDS/STDs. Violence against women was expected in relation to a positive test result. There is a need for better educative effort on the modes of transmission and prevention of HIV, also in 'low risk' populations. PMID- 10471107 TI - Male polyurethane condoms do not enhance brief HIV-STD risk reduction interventions for heterosexually active men: results from a randomized test of concept. AB - The current study examined the effects of a brief HIV risk reduction intervention for men. Participants were recruited (n = 108) from an urban public health clinic and randomly assigned to one of 3 experimental intervention conditions: (1) 3 h HIV-STD risk reduction behavioural skills building latex condom intervention; (2) the same 3 h skills workshop but focused on male polyurethane condoms; or (3) a 3 h HIV education workshop. Results showed all 3 interventions increased AIDS knowledge and positively influenced attitudes, but men who received the polyurethane condom skills intervention were more likely to request condoms at follow ups. In addition, men who received either of the 2 skills interventions evidenced increased condom use at the one-month follow up, with no differences at 3-month follow up. This study is among the first to test a brief HIV risk reduction intervention for men and is the first to test whether polyurethane male condoms enhance HIV risk reduction efforts. PMID- 10471108 TI - An audit of diagnostic coding in genitourinary medicine clinics. Trent Region Genitourinary Medicine Audit Group. AB - The KC60 diagnostic code information provides an epidemiological monitor of sexually transmitted infection (STI) and a means of standardizing clinical workload in genitourinary medicine (GUM) departments. We aimed to assess the coding process and its uniformity within the Trent region by means of a confidential self-administered coding exercise with simulated case presentations. The correct coding for STIs ranged from 59.4% to 100% in different scenarios. Difficulty was identified in the coding of vulvitis and balanitis where no organism was isolated, as represented by a wide range of codes. HIV pre-test counselling without testing was coded by only 57.8% of respondents. Over 95% indicated correctly the first hepatitis B vaccine dose but 21% failed to code once only for the course. Coding practice also varied within individual sites with a 21-100% discordance. In parts diagnostic accuracy was good but there were areas of non-uniformity both within sites and cross-regionally. Regional discussions have resulted in increased training opportunities and guidelines have been developed to increase uniformity and achieve consensus in uncertain areas. PMID- 10471109 TI - Nandrolone decanoate; use in HIV-associated lipodystrophy syndrome: a pilot study. PMID- 10471111 TI - Transport of fatty acids across membranes by the diffusion mechanism. AB - In early research on fatty acid transport, passive diffusion seemed to provide an adequate explanation for movement of fatty acids through the membrane bilayer. This simple hypothesis was later challenged by the discovery of several proteins that appeared to be membrane-related fatty acid transporters. In addition, some biophysical studies suggested that fatty acids moved slowly through the simple model membranes (phospholipid bilayers), which would provide a rationale for protein-assisted transport. Furthermore, it was difficult to rationalize how fatty acids could diffuse passively across the bilayer as anions. Newer studies have shown that fatty acids are present in membranes in the un-ionized as well as the ionized form, and that the un-ionized form can cross a protein-free phospholipid bilayer quickly. This flip-flop mechanism has been validated in cells by intracellular pH measurements. The role of putative fatty acid transport proteins remains to be clarified. PMID- 10471110 TI - Molecular aspects of fatty acid transport: mutations in the IYTSGTTGXPK motif impair fatty acid transport protein function. AB - The murine fatty acid transport protein (FATP) facilitates uptake of long chain fatty acids (LCFAs) when expressed in mammalian cells. FATP's sequence contains a highly conserved motif, IYTSGTTGXPK, also found in a number of proteins known to interact with ATP. To explore the role of this motif, we independently mutated the central serine (serine 250) and threonine (threonine 252) residues in this motif and assessed the effects of these mutations on FATP function. When expressed in fibroblasts, the FATP mutants demonstrated impaired LCFA import and impaired binding of [alpha-32P]8-azido-ATP (azido-ATP) compared with wild-type FATP. These results suggest that serine 250 and threonine 252 are critical for FATP function and that the mechanism of action of FATP involves nucleotide binding which is dependent on these residues. PMID- 10471112 TI - Protein acylation in normoxic and ischemic/reperfused cardiac tissue. AB - In addition to a prominent role in tissue energy conversion, fatty acids are involved in signal transduction and modulation of cellular protein localization and function. The latter is accomplished by acylation of specific cellular proteins. In the present study the amount of fatty acyl moieties covalently bound to cardiac proteins and the effect of myocardial ischemia and reperfusion on the degree and relative fatty acyl composition of cardiac proteins have been investigated in isolated rat hearts. In the normoxic heart about 0.32% of the cellular fatty acyl pool is covalently bound to proteins. Approximately 90% of these fatty acyl chains are thio-esterified, whereas a relatively minor part is attached to cardiac proteins through amide linkage. Thio-esterified fatty acyl chains are derived from palmitic, stearic, oleic, linoleic, arachidonic and docosahexaenoic acid. In contrast, amide linked protein acylation shows a preference for myristic acyl chains. Acute ischemia and reperfusion inflicted upon the isolated rat heart did enhance significantly the content of (unesterified) fatty acids, but did neither affect the degree of protein acylation nor the relative fatty acyl composition of acylated proteins in cardiac tissue. PMID- 10471113 TI - Functional and electrophysiologic effects of polyunsaturated fatty acids on exictable tissues: heart and brain. AB - It has been shown in animals and probably in humans, that n-3 polyunsaturated fatty acids (PUFAs) are antiarrhythmic. The free PUFAs stabilize the electrical activity of isolated cardiac myocytes by inhibiting sarcolemmal ion channels, so that a stronger electrical stimulus is required to elicit an action potential and the relative refractory period is markedly prolonged. This appears at present to be the probable major antiarrhythmic mechanism of the PUFAs. They similarly inhibit the Na+ and Ca2+ currents in rat hippocampal neurons which results in an increase in the electrical threshold for generalized seizures using the cortical stimulation model in rats. PMID- 10471114 TI - Fatty acid ethyl esters: current facts and speculations. AB - Increasing evidence indicates that fatty acid ethyl esters (FAEE) play a role in ethanol-induced organ damage and may serve as long-term markers of ethanol intake. This report summarizes the current knowledge on the toxicity of FAEE, the enzymes associated with FAEE synthesis, FAEE as fatty acid supplements, the in vivo degradation of orally ingested FAEE and FAEE as markers of ethanol intake. A list of major unanswered questions in each of these categories is also included. PMID- 10471115 TI - Polyunsaturated fatty acid biosynthesis: a microsomal-peroxisomal process. AB - The synthesis of 22-carbon fatty acids, with their first double bond at position 4, requires the participation of enzymes in both peroxisomes and the endoplasmic reticulum as well as the controlled movement of fatty acids between these two cellular compartments. It has been observed that there is generally an inverse relationship between rates of peroxisomal beta-oxidation vs those for the microsomal esterification of fatty acids into 1-acyl-sn-glycero-3-phosphocholine. With a variety of different substrates it was found that when a fatty acid is produced in peroxisomes, with its first double bond at position 4, its preferred metabolic fate is to move to microsomes for esterification rather than to serve as a substrate for continued degradation. The required movement, and the associated reactions, in peroxisomes and microsomes is not restricted to the synthesis of 4,7,10,13,16-docosapentaenoic acid and 4,7,10,13,16,19 docosahexaenoic acid. When microsomes and peroxisomes were incubated with NAD, NADPH and malonyl-CoA it was found that 6,9,12-octadecatrienoic acid was metabolized to linoleate. Collectively our findings suggest that there may be considerably more recycling of fatty acids between peroxisomes and the endoplasmic reticulum than was previously recognized. PMID- 10471116 TI - Human very long-chain acyl-CoA synthetase and two human homologs: initial characterization and relationship to fatty acid transport protein. AB - Several human genes with a high degree of homology to rat very long-chain acyl CoA synthetase (rVLCS) and mouse fatty acid transport protein (mFATP) were identified. Full-length cDNA clones were obtained for three genes, and predicted amino acid sequences were generated. Initial characterization indicated that one gene was most likely hVLCS, the human ortholog of rVLCS. The other two (hVLCS-H1 and hVLCS-H2) were more closely related to rVLCS than to mFATP. Phylogenetic analysis of amino acid sequences confirmed that hVLCS-H1 and hVLCS-H2 were evolutionarily closer to VLCSs than FATPs. Alignment of predicted amino acid sequences of human, rat and mouse VLCSs and FATPs revealed the existence of two highly conserved motifs. While one motif is also present in long-chain acyl-CoA synthetases, the other serves to distinguish the VLCS/FATP family from the long chain synthetase family. Elucidation of the biochemical functions of all VLCS/FATP family members should provide new insights into cellular fatty acid metabolism. PMID- 10471117 TI - Omega-3 fatty acids and bipolar disorder: a review. AB - The important role of the omega-3 fatty acids in the pathophysiology and treatment of bipolar disorder is now supported by a substantial body of indirect and direct evidence. This paper will describe the clinical and pharmacological features of bipolar disorder, review the available data regarding omega-3 fatty acids in bipolar disorder and provide recommendations for future research. PMID- 10471119 TI - Multiple mechanisms for polyunsaturated fatty acid regulation of hepatic gene transcription. AB - Dietary polyunsaturated fatty acids (PUFA) have profound effects on hepatic gene transcription leading to significant changes in lipid metabolism. Highly unsaturated n-3 PUFA suppress the transcription of genes encoding specific lipogenic enzymes and induce the expression of genes encoding specific enzymes involved in peroxisomal and microsomal fatty acid oxidation. Our studies have shown that fatty acid effects on hepatic gene expression may involve at least three distinct pathways. One pathway involves peroxisome proliferator-activated receptor (PPARalpha), a fatty acid activated nuclear receptor. PPARalpha is required for the PUFA induction of mRNAs encoding enzymes involved in fatty acid oxidation. However, PPARalpha is not required for PUFA suppression of mRNAs encoding proteins involved in lipogenesis. A second pathway involves prostanoids. In cultured 3T3-L1 adipocytes, cyclooxygenase derived 20:4 n-6 metabolites, like PGE2, suppress mRNAs encoding proteins involved in lipogenesis. However, in hepatic parenchymal cells, 20:4 n-6 suppression of lipogenic gene expression does not require a cyclooxygenase. Nevertheless, PGE2 and PGF2alpha suppress hepatic lipogenic gene expression. 20:4 n-6 cyclooxygenase products can arise from non parenchymal cells and through a paracrine control process act on a G-protein linked receptor signaling cascade to suppress lipogenic gene expression. The fact that n-3 and n-6 PUFA suppression of lipogenic gene expression does not require PPARalpha or cyclooxygenase activity indicates the presence of a third pathway for the control of hepatic gene transcription. These studies indicate that the pleiotropic effects of PUFA on hepatic lipid metabolism cannot be attributed to a single regulatory mechanism. PMID- 10471118 TI - The role of the peroxisome proliferator-activated receptor alpha (PPAR alpha) in the control of cardiac lipid metabolism. AB - The postnatal mammalian heart uses mitochondrial fatty acid oxidation (FAO) as the chief source of energy to meet the high energy demands necessary for pump function. Flux through the cardiac FAO pathway is tightly controlled in accordance with energy demands dictated by diverse physiologic and dietary conditions. In this report, we demonstrate that the lipid-activated nuclear receptor, peroxisome proliferator-activated receptor alpha (PPARalpha), regulates the expression of several key enzymes involved in cardiac mitochondrial FAO. In response to the metabolic stress imposed by pharmacologic inhibition of mitochondrial long-chain fatty acid import with etomoxir, PPARa serves as a molecular 'lipostat' factor by inducing the expression of target genes involved in fatty acid utilization including enzymes involved in mitochondrial and peroxisomal beta-oxidation pathways. In mice lacking PPARalpha (PPARalpha-/- mice), etomoxir precipitates a cardiac phenotype characterized by myocyte lipid accumulation. Surprisingly, this metabolic regulatory response is influenced by gender as demonstrated by the observation that male PPARalpha-/- mice are more susceptible to the metabolic stress compared to female animals. These results identify an important role for PPARalpha in the control of cardiac lipid metabolism. PMID- 10471120 TI - Coordinate induction of peroxisomal acyl-CoA oxidase and UCP-3 by dietary fish oil: a mechanism for decreased body fat deposition. AB - Rats fed dietary fats rich in 20- and 22-carbon polyenoic fatty acids deposit less fat and expend more energy at rest than rats fed other types of fats. We hypothesized that this decrease in energetic efficiency was the product of: (a) enhanced peroxisomal fatty acid oxidation and/or (b) the up-regulation of genes encoding proteins that were involved with enhanced heat production, i.e. mitochondrial uncoupling proteins (UCP-2, UCP-3) and peroxisomal fatty acid oxidation proteins. Two groups of male Fisher 344 rats 3-4 week old (n=5 per group) were pair fed for 6 weeks a diet containing 40% of its energy fat derived from either fish oil or corn oil. Epididymal fat pads from rats fed the fish oil diet weighed 25% (P < 0.05) less than those found in rats fed corn oil. The decrease in fat deposition associated with fish oil ingestion was accompanied by a significant increase in the abundance of skeletal muscle UCP-3 mRNA. The level of UCP-2 mRNA skeletal muscle was unaffected by the type of dietary oil, but the abundance of UCP-2 mRNA in the liver and heart were significantly lower (P < 0.05) in rats fed fish oil than in rats fed corn oil. In addition to inducing UCP 3 expression, dietary fish oil induced peroxisomal acyl-CoA oxidase gene expression 2-3 fold in liver, skeletal muscle and heart. These data support the hypothesis that dietary fish oil reduces fat deposition by increasing the expression of mitochondrial uncoupling proteins and increasing fatty acid oxidation by the less efficient peroxisomal pathway. PMID- 10471121 TI - Regulation of alpha-foetoprotein gene expression by fatty acids and fibrates. AB - Alpha-foetoprotein (AFP), the major plasma protein in the foetus, is mainly synthesized by yolk sac and foetal liver. It binds polyunsaturated fatty acids and probably controls their metabolism and action. We investigated the effects of fatty acids and fibrates on expression of the AFP gene using two complementary approaches. Treatment with 5-8-11-14 eicosatetraynoic acid (ETYA), an analogue of arachidonic acid, specifically led to lower AFP mRNA levels in cultured rat yolk sac explants whereas treatment with palmitic or oleic acid did not. Clofibric acid and fenofibrate also gave lower AFP mRNA levels. Transient transfection experiments with HepG2 hepatoma cells showed that ETYA and clofibric acid decreased the transcriptional activity of the 7 kb regulatory region of the rat AFP gene. The 330 bp AFP promoter was identified as a target for these down regulating effects. PMID- 10471122 TI - Differential effects of long chain n-3 fatty acids on the expression of PGH synthase isoforms in bovine aortic endothelial cells. AB - Primary cultures of bovine aortic endothelial cells were used at confluency to evaluate the effect of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids on the expression of both the constitutive and inducible isoforms of PGH synthase (PGHS), PGHS-1 and PGHS-2, respectively. After a 22 h period enrichment of cell lipid stores with each fatty acid, the expression of PGH synthase isoforms was measured by western blotting. EPA and DHA, but not oleate, significantly decreased the immunoreactive PGHS-1 and to a similar extent the corresponding mRNA, as measured by northern blotting. Studies on mRNA stability failed to show any difference between DHA-enriched and control cells, indicating that the decreased expression observed was likely from transcriptional origin. Under the enrichment conditions, EPA and DHA, but not oleate, moderately but significantly induced an oxidative stress as judged by malondialdehyde formation. Interestingly, hydrogen peroxide was able to mimic the effect of EPA and DHA in decreasing the expression of PGHS-1. On the other hand, the PMA-induced PGHS-2 expression could be potentiated by cell pre-enrichment with DHA, whereas hydrogen peroxide alone could induce such an expression. We conclude that the long chain n 3 fatty acids EPA and DHA may differently affect the expression of PGH synthase isoforms, possibly via an oxidative stress. PMID- 10471123 TI - Arachidonic and docosahexaenoic acids differentially affect the expression of fatty acyl-CoA oxidase, protein kinase C and lipid peroxidation in HepG2 cells. AB - Arachidonic (AA) and docosahexaenoic (DHA) acids (5-20 microM), when supplemented to human hepatoma HepG2 cells, which are depleted in these long-chain polyunsaturated fatty acids in conventional culture conditions, enhance the expression of acyl-CoA oxidase (ACOX), the first enzyme in the peroxisomal beta oxidation cycle. DHA is effective at lower concentrations (at 5 microM) and to a greater extent (about 60% increment) than AA (about 40%) at 20 microM. Protein kinase C (PKC) appears to be involved in the activity of AA on ACOX, but not in that of DHA, since only the effect of AA is prevented by the PKC inhibitor Staurosporine, and since a remarkable elevation of the PKC activator diacylglycerol occurs only after AA supplementation. AA also induces elevation of lipoperoxides, favoured by the relative vitamin E deficiency occurring in cultured cells, and this effect, which is prevented by supplementation of the vitamin, may contribute to PKC activation. PMID- 10471124 TI - Human peripheral blood T lymphocyte proliferation after activation of the T cell receptor: effects of unsaturated fatty acids. AB - Oils enriched in certain polyunsaturated fatty acids suppress joint pain and swelling in rheumatoid arthritis patients with active synovitis. Because T lymphocyte activation is important for propagation of joint tissue injury in patients with rheumatoid arthritis, we examined the effects of fatty acids added in vitro on proliferation of human T lymphocytes stimulated with monoclonal antibodies to CD3 and CD4. Unsaturated fatty acids reduced T cell proliferation in a dose dependent manner (dihomogammalinolenic acid > gammalinolenic acid > eicosapentaenoic acid > arachidonic acid). Removal of fatty acids from cultures before cell stimulation did not change the effects, but addition of fatty acids after cell stimulation failed to reduce T cell responses. The saturated palmitic acid did not influence T cell growth. These studies indicate that small changes in cellular fatty acids can have profound effects on early events in T cell signaling and on T cell function. PMID- 10471126 TI - Prostacyclin as a critical prostanoid in adipogenesis. AB - Adipose cell differentiation from adipoblasts to preadipose and to adipose cells is a multistep process. Terminal differentiation of preadipose cells expressing early markers to adipose cells expressing late and very late markers and accumulating triacylglycerol requires a combination of circulating and locally produced hormones. Prostacyclin (PGI2), one of the major metabolites of arachidonic acid in adipose tissue, has been shown to exert autocrine and paracrine adipogenic effects in vitro. As discussed herein, multiple arguments support the proposition that PGI2 is a key prostanoid involved in adipogenesis. PMID- 10471125 TI - Role of peroxisomal oxidation in the conversion of arachidonic acid to eicosatrienoic acid in human skin fibroblasts. AB - Human skin fibroblasts converted [5,6,8,9,11,12,14,15-3H]arachidonic acid ([3H]20:4) to eicosatrienoic acid (20:3), but appreciable amounts of radiolabeled 20:3 were not detected in corresponding incubations with [1-(14)C]20:4. This indicates that the main pathway for synthesizing 20:3 from arachidonic acid in the fibroblast involves oxidative removal of the carboxyl group of arachidonic acid. Fibroblasts deficient in long-chain acyl coenzyme A dehydrogenase (LCAD) converted [3H]20:4 to [3H]20:3. However, Zellweger fibroblasts that are deficient in peroxisomal fatty acid oxidation did not, indicating that the oxidative removal of the carboxyl group occurs in the peroxisomes. [3H]Hexadecatrienoic acid (16:3) was the main product that accumulated when [3H]20:4 was incubated with normal, LCAD deficient, and very long-chain acyl coenzyme A dehydrogenase (VLCAD) deficient fibroblasts, but Zellweger fibroblasts did not form this product. Normal fibroblasts converted [3H]16:3 to radiolabeled 20:3 and arachidonic acid. These findings suggest that some of the 16:3 produced from arachidonic acid by peroxisomal beta-oxidation can be recycled and that this recycling process constitutes a novel pathway for the conversion of arachidonic acid to 20:3 in human fibroblasts. PMID- 10471127 TI - Carbon recycling into de novo lipogenesis is a major pathway in neonatal metabolism of linoleate and alpha-linolenate. AB - Recent reports indicate that recycling of the beta-oxidized carbon skeleton of linoleate and alpha-linolenate into newly synthesized cholesterol and fatty acids in the brain is quantitatively significant in both suckling rats and pre- and postnatally in rhesus monkeys. The recycling appears to occur via ketones which are not only readily produced from these 18 carbon polyunsaturates but are also the main lipogenic precursors for the developing mammalian brain. Since the neonatal rat brain appears not to acquire cholesterol or long chain saturated or monounsaturated fatty acids from the circulation, ketones and ketogenic precursors seem to be crucial for normal brain synthesis of these lipids. Cholesterol is plentiful in brain membranes and it has also been discovered to be the essential lipid adduct of the 'hedgehog' family of proteins, the appropriate expression of which determines normal embryonic tissue patterning and neurological development. Insufficient cholesterol or inappropriate expression of 'sonic hedgehog' has major adverse neurodevelopmental consequences typified in humans by Smith-Lemli-Optiz syndrome. Hence, we propose that the importance of alpha-linolenate and linoleate for normal neural development arises not only from being precursors to longer chain polyunsaturates incorporated into neuronal membranes but, perhaps equally importantly, by being ketogenic precursors needed for in situ brain lipid synthesis. PMID- 10471128 TI - Does high polyunsaturated free fatty acid level at the feto-maternal interface alter steroid hormone message during pregnancy? AB - Polyunsaturated fatty acids (PUFA) are important in pregnancy, fetal development and parturition. We measured free fatty acids (FFA), albumin and alpha fetoprotein (AFP) in the maternal and fetal circulations of women undergoing elective Caesarean section at term. We also studied the impact of PUFAs on estrogen (ER) and progesterone receptors (PR) binding properties in vitro in the myometria of pregnant women and ex vivo in human myometrial cells in culture. FFA in intervillous blood (I) (feto-maternal interface) and maternal peripheral blood (M) were similar, while those in the umbilical vein (V) and arteries (A) were 2-4 fold lower (P<0.001). PUFA levels were low in M and 3 fold higher in I, A and V (P< 0.001); consequently C20:4 and C22:6 were most abundant in intervillous space. Albumin was uniformly distributed throughout the maternal-fetal unit, but there was a transplacental gradient in AFP. The AFP in the intervillous space had a special conformation (less immuno-reactive, more anionic), suggesting loading with PUFA. Physiological concentrations of C20:4 stimulated estradiol binding, but inhibited progestin binding. C20:4 inhibited progesterone binding by decreasing the number of binding sites, with no change in apparent affinity, in vitro in myometrial tissue and ex vivo in myometrial cells. Thus PUFA may modulate the steroid hormone message, so that the high C20:4 concentration at the maternal-fetal interface at term may help amplify the estrogen signal and inhibit the progesterone signal. PMID- 10471129 TI - Plasma aldosterone, plasma lipoproteins, obesity and insulin resistance in humans. AB - Aldosterone production in vitro can be affected by many hormones, autacoids, ions, and lipids, but regulation in humans is incompletely understood. We measured plasma aldosterone in adult subjects with a wide range of obesity and insulin resistance. Aldosterone levels correlated with measures of visceral obesity in one predominantly male cohort and in the women of a second cohort. In the same subjects, aldosterone correlated with insulin resistance. Aldosterone also correlated with plasma cortisol in men and women, and with DHEA-S in women. The data suggested that visceral fat stimulates adrenal steroidogenesis. We found that certain fatty acids stimulated aldosterone production in vitro by rat adrenal cells incubated with rat hepatocytes, but not adrenal cells alone. The results suggested that fatty acids from visceral adipocytes induce hepatic formation of an adrenal secretagogue. This may explain the correlation of plasma steroids with visceral obesity. Aldosterone may contribute to vascular diseases that complicate obesity. PMID- 10471130 TI - In vivo conversion of linoleic acid to arachidonic acid in human adults. AB - Human adults are shown to be capable of conversion of linoleic acid (LA, 18:2 n 6) to arachidonic acid (AA, 20:4 n-6) in vivo. It is confirmed that they can also convert alpha-linolenic acid (LNA, 18:3 n-3) to eicosapentaenoic acid (EPA, 20:5 n-3) and to docosahexaenoic acid (DHA, 22:6 n-3) in vivo. The time course and the maximal response for these processes during the first week after a single dose of the 18-carbon precursor is described. A stable-isotope method in which the protons of the C17 and C18 carbons are substituted with deuterium atoms is used in order to provide for a safe method for the study of human metabolism. High sensitivity and selectivity of detection is assured with negative ion, gas chromatography/mass spectrometry analysis. It is clear that human adults on an ad lib diet carry out EFA metabolism in vivo. PMID- 10471131 TI - Vascular effects of non-esterified fatty acids: implications for the cardiovascular risk factor cluster. AB - Insulin resistance emerges as a central component of the risk factor cluster and is a likely contributor to vascular disease independently of traditional risk factors such as hypertension and diabetes mellitus. However, the intermediary mechanisms by which atherosclerosis is accelerated among patients with the insulin resistance syndrome remain inadequately defined. Most of the attention has centered on hyperinsulinemia and defects of insulin-mediated glucose disposal. However, we observed that obese hypertensive patients have elevated plasma concentrations of non-esterified fatty acids (NEFAs), including oleic acid, which are highly resistant to suppression by insulin. Resistance to insulin's fatty acid lowering action correlate with blood pressure in obese subjects independently of defects in glucose disposal. This observation raises the possibility that NEFAs have biologically significant effects on the cardiovascular system. In fact, oleic acid impairs nitric oxide synthase activity and endothelium-dependent vasorelaxation in vitro. Moreover, raising NEFAs in normal human volunteers to levels observed in obese hypertensive patients impairs lower extremity endothelium-dependent vasodilation and augments local and systemic vascular alpha1-adrenoceptor reactivity in normal volunteers. Thus, raising NEFAs replicates in healthy subjects important functional vascular changes implicated in the hypertension and atherosclerosis observed in patients with the risk factor cluster. At a molecular level, experiments in cultured vascular smooth muscle cells demonstrate that oleic acid activates a mitogenic signaling cascade which includes protein kinase C, reactive oxygen species and extracellular signal-regulated kinases. Each of these signaling events has been implicated in the structural and functional vascular changes which accompany the risk factor cluster. Collectively, these observations raise the possibility that fatty acids contribute to functional and structural vascular changes among insulin-resistant individuals. A better understanding of the signaling mechanisms by which NEFAs exert their vascular effects may facilitate novel and more effective therapeutic approaches to managing the cardiovascular risk factor cluster. PMID- 10471132 TI - Evolutionary aspects of omega-3 fatty acids in the food supply. AB - Information from archaeological findings and studies from modern day hunter gatherers suggest that the Paleolithic diet is the diet we evolved on and for which our genetic profile was programmed. The Paleolithic diet is characterized by lower fat and lower saturated fat intake than Western diets; a balanced intake of omega-6 and omega-3 essential fatty acids; small amounts of trans fatty acids, contributing less than 2% of dietary energy; more green leafy vegetables and fruits providing higher levels of vitamin E and vitamin C and other antioxidants than today's diet and higher amounts of calcium and potassium but lower sodium intake. Studies on the traditional Greek diet (diet of Crete) indicate an omega 6/omega-3 ratio of about 1/1. The importance of a balanced ratio of omega-6:omega 3, a lower saturated fatty acid and lower total fat intake (30-33%), along with higher intakes of fruits and vegetables leading to increases in vitamin E and C, was tested in the Lyon Heart study. The Lyon study, based on a modified diet of Crete, confirmed the importance of omega-3 fatty acids from marine and terrestrial sources, and vitamin E and vitamin C, in the secondary prevention of coronary heart disease, and cancer mortality. PMID- 10471133 TI - Lipid metabolism, human evolution and schizophrenia. AB - There are only small genetic differences between humans and the great apes. Yet these differences must be very important. Major known differences include the accumulation of subcutaneous fat, the expansion of breasts and buttocks, the growth of the brain and the connectivity of neurons. All these involve lipid metabolism yet, because fat leaves no fossils, lipids are rarely mentioned in discussions of human evolution. This paper attempts to identify some candidate areas of lipid metabolism which may be important in human evolution. It draws attention to abnormalities in phospholipid metabolism in schizophrenia and suggests that these may have proved important in enhancing brain connectivity in the later stages of evolution of modern humans. PMID- 10471134 TI - Ovarian tissue graft: from animal experiment to practice in the human. PMID- 10471135 TI - Criteria for selective screening of cervical Chlamydia trachomatis infection in women attending private gynecology practices. AB - OBJECTIVE: To develop a screening strategy for Chlamydia trachomatis in private gynecology practices. STUDY DESIGN: Forty-six gynecologists in the greater Paris area systematically screened all consecutive female attenders during one week. Endocervical swabs were tested by enzyme immunoassay, and, if positive, was further confirmed by direct fluorescent antibody test. RESULTS: Overall (out of 1893 women tested), the chlamydia prevalence was 0.8% (95% confidence interval: 0.32-1.2%). Among women younger than 21 years, the prevalence was 5.2% (95%, CI 0.8-9.6). Having had a new sexual partner in the last year was associated with chlamydial infection, independent of age. However, half of the infected women had not had a new partner. CONCLUSION: Even in this low risk population, the prevalence was high among young women. Most of these infections would probably not have been detected. We developed a strategy which would have detected 81% of chlamydia positive women by testing only one third of the patients. PMID- 10471136 TI - Prevalence of hydronephrosis in patients with genital prolapse. AB - OBJECTIVE: To determine the prevalence of hydronephrosis in patients with genital prolapse and evaluate contributing factors. METHODS: Retrospective study of the records of 189 patients with pelvic organ prolapse who underwent preoperative renal imaging studies. RESULTS: Of the 189 patients 31 (17.4%) had hydronephrosis; in 20 (10.6%) patients the hydronephrosis was mild, in 7 (3.7%) it was moderate and in 4 (2.7%) it was severe. The patients with hydronephrosis were older (mean age 68+/-9.5 SD vs. 60.5+/-10.8 SD, P<0.001) had a higher creatinine levels (0.84+/-0.4 SD vs. 0.78+/-0.3 SD P<0.005) and had a higher degree of uterine prolapse (mean 2.6+/-0.9 SD vs. 1.1+/-1.2 SD, P<0.005). After adjusting for age and type of prolapse, only patients with uterine prolapse remained significantly more likely to suffer from hydronephrosis (adjusted odds ratio 1.9, 95% CI 1.1, 3.2). CONCLUSION: The prevalence of hydronephrosis in patients with genital prolapse is appreciable and is related primarily to degree of uterine prolapse. PMID- 10471137 TI - Brain damage and hypoxia in an ovine fetal chronic cocaine model. AB - OBJECTIVE: To assess the development of brain damage in an ovine fetal chronic cocaine model. To evaluate the effect of isolated hypoxic tests on this model and to correlate hemodynamic findings (brain-sparing effect) following fetal hypoxia and the occurrence of brain damage. STUDY DESIGN: Fifteen ewes were divided into a control group (n=7) and a cocaine treated group (n=8). From day 65 to day 134 the cocaine treated animals received a daily (5 days per week) intramuscular injection (2 mg/kg cocaine) and the control animals a placebo injection (2 ml of isotonic solution). Both groups underwent hypoxic tests (cord compression (3 min) and aortic compression (1 min)) at 90 and 134 days. In addition, anesthesia for magnetic resonance imaging (MRI) examination was carried out at 125 days. Fetal blood samples were collected during both series of hypoxic tests and the cerebral and umbilical flows were monitored by Doppler. Samples from 25 brains (control n = 10; cocaine n= 15) were processed for light and electron microscopic examination. Quantification of brain damage was done on semithin sections from six areas of cortex and germinal matrix on each fetus. RESULTS: Similar forms of brain damage (selective neuronal loss limited to the parasaggital cortex, striatum, hippocampus and Purkinje cells) was present in both groups but lesions were more frequent in the cocaine treated group as shown by quantitative analysis for the proportion of abnormal capillaries (65% vs. 35%), capillary edema (61% vs. 34%) and abnormal neurons showing delayed neuronal degeneration (DND) (66% vs. 36%) in the cocaine and control group respectively. There was no significant difference in immunoreactivity for glial fibrillary acidic protein (GFAP) but it was more marked in the cerebellum of cocaine treated animals. Fetal blood samples showed a moderate sustained hypoxia and Doppler findings demonstrated the presence of a brain sparing effect associated with increased uterine and umbilical vascular resistance in the cocaine treated group. Nevertheless, the amplitude of the heart rate increase and cerebral dilatation was significantly lower in the cocaine treated animals. CONCLUSION: This ovine fetal chronic cocaine model showed the presence of brain damage. Cocaine treatment seems to potentiate the effect of the hypoxic tests. Independent of the cause, the brain damage developed in the presence of brain sparing effect, strongly suggesting that this phenomenon is a sign of a pathological fetal condition and no guarantee that it will prevent tissue damage. PMID- 10471138 TI - Perinatal mortality in twin pregnancies: a 3-year analysis in Seine Saint-Denis (France). AB - OBJECTIVES: Our purpose was to evaluate perinatal mortality in twins and to investigate factors associated with this mortality. STUDY DESIGN: A prospective study on the perinatal mortality was performed in the department of Seine-Saint Denis. Out of all the perinatal deaths, we have retrospectively isolated those arising from twin pregnancies. RESULTS: There were 54 twin pregnancies complicated with perinatal death. The perinatal mortality rate in twin pregnancy was 78.0 per 1000 twin babies delivered. Out of 86 twin deaths, 38 (44.2%) were born before 28 weeks gestation and out of 82 twin perinatal deaths, 37 (45.1%) weighed less than 1000 g. Chorionicity was recorded in 44 twin pairs: 21 (47.7%) were dichorionic and 23 (52.3%) monochorionic. Finally, out of 48 twin sets there were four (8.3%) monoamniotic pregnancies. CONCLUSIONS: The present data show that extreme prematurity represents nearly half of perinatal mortality in twins. This study indicates also a significant proportion of monochorionic placentation among twin pregnancies with poor outcome. PMID- 10471139 TI - Postpartum oral glucose tolerance tests in mothers of macarosomic infants: inadequacy of current antenatal test criteria in detecting prediabetic state. AB - OBJECTIVE: To assess the presence of subtle carbohydrate metabolism abnormalities in otherwise healthy mothers who have given macrosomic birth by utilizing postpartum oral glucose tolerance test (PPOGTT). STUDY DESIGN: Prospective controlled study enrolled gestational diabetic women (GDM, n=10), mothers with macrosomic infants (MwMIs, n=62) and controls (n=50). RESULTS: Receiver operating characteristic (ROC) curve analysis revealed that incremental 1-h+2-h PPOGTT value >111 mg/dl had a sensitivity of 80% and specificity of 78% in predicting antecedent diabetes. PPOGTT results were positive in 53.2% of MwMIs and 28% of controls (P<0.01). Maternal low-density lipoprotein and triglyceride levels, 50 gram glucose challenge test (50 g GCT) values and neonatal weight were the significant predictors of PPOGTT results. ROC analyses suggested that threshold of 50 g GCT should be lowered in order to better predict subjects with both macrosomia and positive PPOGTT. CONCLUSION: PPOGTT may identify a subset of women with macrosomic infants who have metabolic alterations of a prediabetic state. The discrepancies between antenatal and postpartum tests may reflect the need for redefinition of currently utilized criteria in screening and diagnosis of GDM. PMID- 10471140 TI - Elevated soluble adhesion molecules in women with pre-eclampsia. Do cytokines like tumour necrosis factor-alpha and interleukin-1beta cause endothelial activation. AB - OBJECTIVE: The purpose of the present study was to evaluate the clinical significance of tumour necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) for endothelial cell activation in pre-eclampsia. Therefore, we determined and compared the correlations between these cytokines and circulating adhesion molecules in the sera of pre-eclamptic pregnant women, normotensive pregnant women and nonpregnant women. METHODS: The soluble adhesion molecules VCAM-1, ICAM-1, E-selectin, and P-selectin were determined in the serum of 38 pre eclamptic pregnant women and 40 normotensive pregnant and nonpregnant controls using ELISA-techniques. We correlated these serum concentrations with the serum levels of TNF-alpha and IL-1beta, respectively, also determined by ELISA. RESULTS: Elevated serum levels of VCAM-1 and E-selectin could be detected in pre eclamptic patients, with and without HELLP-syndrome. In contrast, no increased serum concentration of ICAM-1, P-selectin, TNF-alpha and IL-1beta were found in these patients. While significant correlation between VCAM-1 and E-selectin could be determined (r=0.604; p<0.001) no unambiguous correlations, however, were found between TNF-alpha or between IL-1beta and the examined adhesion molecules or the selectins. CONCLUSIONS: In contrast to in vitro investigations on cultured umbilical vein endothelium, our experimental results indicate that the cytokines TNF-alpha and IL-1beta can not explain endothelial cell activation, and that their measurement in serum is not useful for the detection of pre-eclampsia. PMID- 10471141 TI - School performance and behaviour in extremely preterm growth-retarded infants. AB - OBJECTIVE: To describe school performance and behaviour of extremely preterm, growth-retarded infants. DESIGN: Cohort study at two tertiary care centres. Included were all surviving, singleton infants (N= 127) with fetal growth retardation due to placental insufficiency. All were delivered by caesarean section because of signs of fetal distress before the beginning of labour at a gestational age of 26 to 32 weeks during the years 1984-1989. Main outcome measures were special education, mainstream education below the appropriate age level and behaviour according to attention-deficit hyperactivity criteria at school age (4 1/2-10 1/2 yrs). The children were divided into two subgroups according to age at follow-up (> or =7 1/2 and <7 1/2 yr). A logistic regression analysis was performed with special school or repeating a grade and behavioural disturbance as dependent variables and gestational age, birth weight, sex of the infant, neonatal complications (intra cerebral haemorrhage, respiratory distress syndrome, bronchopulmonary dysplasia or sepsis), age category at follow-up and sociodemographic factors as independent variables. RESULTS: 114 (90%) had a complete follow-up. Special education was found in 14% of the assessed children. More children in the older age group than in the younger age group were placed in special school (20% versus 10%). Behavioural problems were scored in 39% of the assessed children attending mainstream education. Special education was related to neonatal complications (bronchopulmonary dysplasia), behavioural problems to the absence of either parent. CONCLUSION: This specific group of growth-retarded children is at serious disadvantage for adequate performance in school, although the incidence of special education and behavioural problems was comparable to other preterm infants. Both special education and behavioural problems were not related to obstetric variables as gestational age and/or birth weight. PMID- 10471142 TI - The monoamniotic twin: a riskful event. AB - The monoamniotic twin is at risk for high perinatal mortality because of cord accidents. There is no consensus in literature for the obstetric management in these cases. Contradictory results are reported according to fatal complications by cord accidents after 32 weeks gestation and safety of vaginal delivery. Recently two patients were admitted to our hospital with monoamniotic twin pregnancies and a fetal death in the third trimester. On the basis of these cases a review of the literature is presented and a case registry is recommended. PMID- 10471143 TI - Reflex sympathetic dystrophy in pregnancy: nine cases and a review of the literature. AB - OBJECTIVE: To better understand the diagnosis of reflex sympathetic dystrophy of the lower extremities in pregnant women. SUBJECT: Disease analysis using a retrospective series of nine cases and a review of the literature (57 patients and 159 sites of reflex sympathetic dystrophy). RESULTS: This disorder should be considered in any painful pelvic girdle syndrome or lower extremity pain. The hip is involved in 88% of cases. Symptoms develop in the third trimester of pregnancy, between the 26th and the 34th weeks. Magnetic resonance imaging (MRI) provides an early, accurate, and very specific diagnosis, although standard radiography continues to be the first-line diagnostic tool. Fracture occurs in 19% of patients. The etiology and pathophysiology remain unclear, although pregnancy itself appears to play a significant role in this disease. Although locoregional mechanical factors partly explain reflex sympathetic dystrophy. Hypertriglyceridemia appears to be a risk factor. This disorder develops independently, but the conclusion of pregnancy appears to be necessary for cure. Reflex sympathetic dystrophy does not appear to affect the course of the pregnancy. Indications for cesarean delivery remain obstetrical and should be discussed when a fracture is involved. Simple therapeutic management using gentle physical therapy provides rapid and complete recovery in 2-3 months. CONCLUSION: Reflex sympathetic dystrophy during pregnancy remains poorly understood and underestimated. Only joints of the inferior limbs are involved. MRI appears to be the best diagnostic tool. Pathogenesis remains unclear. Fractures are not rare. Treatment should be non-aggressive. PMID- 10471144 TI - Sigmoid colon carcinoma metastatic to the myometrium. AB - Metastases to the uterus from extragenital cancers are significantly rarer than metastases to the ovaries. Of the approximately 200 cases of metastases to the uterus from extragenital cancers that have been reported in the literature, more than half are cases of metastases from breast carcinoma and only 16 are cases of metastases from colorectal carcinoma. A case of isolated metastases restricted to the myometrium of the right uterine comu from sigmoid colon carcinoma, without involvement of the ovaries, is described. The patient who six months previously had surgery for sigmoid colon carcinoma presented with right lower abdominal pain and a palpable mass in the region of the right uterine cornu. The diagnosis of isolated metastases restricted to the myometrium of the right uterine cornu was confirmed by total abdominal hysterectomy and bilateral salpingo-oophorectomy. This case illustrates that a growing uterine mass in a patient with a history of primary extragenital cancer, regardless of whether abnormal uterine bleeding is present or absent, should alert the physician to consider the possibility of uterine metastases. PMID- 10471145 TI - Elevated serum alkaline phosphatase may enable early diagnosis of ovarian cancer. AB - A case of endometrioid ovarian carcinoma associated with elevated levels of serum placental-like alkaline phosphatase (PLAP) is presented. Two and a half years before a final diagnosis was made following explorative laparotomy, an incidental blood test revealed elevated alkaline phosphatase in the patient's serum. A thorough investigation for the source of this elevation was negative. Postoperative immunohistochemical staining of the tumor, showed diffuse stain with PLAP, along with gradual decline to normal values of serum total alkaline phosphatase. It is suggested, that whenever serum alkaline phosphatase is elevated due to unknown reason, an investigation including alkaline phosphatase isoenzymes, serum Ca-125, trans-vaginal pelvic sonogram and even diagnostic laparoscopy, should be considered in a search for early preclinical ovarian cancer. PMID- 10471146 TI - Accumulation and release of iron in polarly and non-polarly cultured trophoblast cells isolated from human term placentas. AB - We investigated the usefulness of membrane grown human term trophoblast cells in transferrin-mediated iron transfer studies. We showed that diferric transferrin is taken up both at the microvillous and at the basal membrane by means of receptor-mediated endocytosis. Uptake from the microvillous side is predominant. This corresponded with a much higher expression of transferrin receptors at the microvillous membrane as compared to the basal one. Iron appeared to accumulate in the cell. Accumulation was higher when transferrin was supplied at the microvillous side. Transfer of iron could not be assessed because uptake of transferrin by the cells was much less than passive diffusion of transferrin through the cell-free filter. The observation of iron accumulation was unexpected for a transfer epithelium. Could it be that part of the iron taken up by the cells is rapidly released whereas the remaining part accumulates? In this case the rate of iron uptake should be higher than the rate of iron accumulation. This question was assessed with non-polarly cultured trophoblast cells. We showed that like in polar cells iron accumulated in ferritin. A new experimental design enabled us to demonstrate that indeed the rate of transferrin-mediated iron is in excess over iron accumulation. We thus provide evidence for a mechanism that enables rapid transfer of iron across the syncytiotrophoblast cell layer. PMID- 10471147 TI - Placental transfer of the thromboxane synthetase inhibitor ridogrel in the late pregnant ewe. AB - OBJECTIVES: To assess the occurrence of placental transfer of the thromboxane synthetase inhibitor ridogrel in the pregnant ewe and to determine its effect on prostanoid levels in the ewe and fetal lamb, on uterine contractility and on maternal and fetal hemodynamics. STUDY DESIGN: Five chronically instrumented pregnant ewes at 122 days of gestation received intravenous infusions of 5 mg/kg/3 h ridogrel and solvent. Maternal and fetal arterial samples were obtained at predetermined intervals to determine concentrations of ridogrel and prostaglandin metabolites TXB2, 6-keto-PGF1alpha, PGF2alpha, and PGE2. Maternal and fetal responses of blood flow and pressures were determined. RESULTS: Fetal ridogrel levels were 25% of maternal concentrations. Ridogrel showed rapid and marked thromboxane synthetase inhibition and augmentation of levels of prostaglandin metabolites. There was no evidence of change in amniotic pressure, uterine blood flow, maternal and fetal blood pressure and heart rate. CONCLUSION: Ridogrel is a potent thromboxane synthetase inhibitor which passes the sheep placenta, does not influence maternal and fetal hemodynamics and uterine contractility, and shows similar antiplatelet activity in the ewe and the fetal lamb. PMID- 10471148 TI - Induction of preeclampsia like phenomena by stimulation of sympathetic nerve with cold and fasting stress. AB - The purpose of this study was to examine the effect of cold-stress, fasting stress and cold plus fasting stress on the sympathetic nerve activity. Pregnant and nonpregnant rats were kept in cold environment (0 degrees C), or fasting condition (12 h), and cold plus fasting condition for 2 weeks. Their plasma corticotrophin releasing factor (CRF), catecholamines, insulin levels, and platelets were measured, and histological examinations were performed. In cold plus fasting stress rats, a significant increased CRF, epinephrine (E), norepinephrine (NE), and insulin levels with decreased platelet count (P<0.0001) were observed compared with control. Histological study revealed that diffused enlarged glomeruli with fibrin deposition in the kidney, hemostasis, ischemic necrosis and fibrin deposition in liver and swelling along with hemorrhagic necrosis in adrenal gland of cold plus fasting stress rats. The biochemical and histological changes in cold plus fasting, cold-stressed or fasting rats were similar to human preeclampsia. The findings observed in cold plus fasting stress rats were more pronounced either than cold-stressed or fasting group. These results demonstrate that cold plus fasting stress is an intense stimulator of sympathetic nervous system than either cold stress or fasting. PMID- 10471149 TI - Bilateral tubal torsion treated by laparoscopy: a case report. AB - The history is described of a patient with bilateral torsion of the fallopian tubes successfully managed by laparoscopy. PMID- 10471150 TI - Villoglandular papillary adenocarcinoma of the uterine cervix. AB - Villoglandular papillary adenocarcinoma of the uterine cervix was recently (1989) described by three main histological features: exophytic proliferation, papillary architecture and mild to moderate cellular atypicality. The authors report a case of villoglandular papillary adenocarcinoma, clinical stage IB, which was peculiar because of its association with a co-existing and simultaneously discovered invasive squamous cell carcinoma. These two patterns were juxtaposed and not intermingled. The patient was treated with radical hysterectomy followed by vaginal radiation therapy. She remains without evidence of recurrence after 12 months of follow-up. Five main clinicopathological features of the villoglandular papillary adenocarcinoma could be stressed: rare histological variant (72 described cases), young age of patients (25-45 years old), superficial stromal invasion, usual association with other tumoral patterns (in situ or invasive adenocarcinoma as well as in situ or invasive squamous cell carcinoma) and excellent prognosis. For selected cases, a conservative surgical approach (cervical conization) was possible. PMID- 10471151 TI - Successful treatment of refractory supraventricular tachycardia by repeat intravascular injection of amiodarone in a fetus with hydrops. AB - We report the case of a fetus with supraventricular tachycardia complicated by congestive heart failure and ascites. After failure of initial transplacental treatment, the injection of amiodarone into the umbilical vein combined with evacuation of ascites achieved conversion to sinus rhythm and restored cardiac function thus allowing pregnancy to go to term. This report suggests that direct fetal therapy by umbilical vein puncture and evacuation of effusions constitutes an effective treatment for supraventricular tachycardias with massive fetal hydrops which do not respond to transplacental treatment. PMID- 10471152 TI - Fetus papyraceous: an unusual cause of obstructed labour. AB - Fetus papyraceous is a relatively rare complication in twin pregnancy. The occurrence of fetus papyraceous is frequently associated with perinatal morbidity in the other twin, making antenatal diagnosis of this condition desirable. Ultrasound detection is not always possible due to anatomical position and technical difficulties. A case of fetus papyraceous, found during Caesarean section for obstructed labour is reported and the implications of antenatal detection are discussed. PMID- 10471153 TI - Max Brodel (1870-1941) and Howard A. Kelly (1858-1943)--urogynecology and the birth of modern medical illustration. AB - In 1894 the German artist Max Brodel came to America and started work with the famous gynecologist Howard A. Kelly. By developing a profound medical knowledge especially in the field of gynecology, and creating new artistic techniques for medical illustration, Brodel revolutionized the appearance of medical literature and founded the profession of medical artist and illustrator. In 1911, Brodel became head of the first 'Department of Art as Applied to Medicine' in the world. The ingenious cooperation of the artist Brodel and the gynecologist Kelly and their influence on modem medical illustration is reflected in this historical article. PMID- 10471154 TI - Direct adenovirus-mediated gene delivery to the temporomandibular joint in guinea pigs. AB - Adenovirus vector system is expected to be useful for direct gene therapy for joint disease. This study first sought to confirm that foreign genes can be transferred to articular chondrocytes in primary culture. Next, recombinant adenovirus vectors harbouring beta-galactosidase gene (LacZ) was injected directly into the temporomandibular joints of Hartley guinea-pigs to clarify the in vivo transfer availability of the adenovirus vectors. Specifically, recombinant adenovirus harbouring LacZ gene (AxlCALacZ) was injected into the upper joint cavities of both mandibular joints of four male 6-week-old Hartley guinea-pigs. Either the same amount of recombinant adenovirus without LacZ gene (Axlw) suspension (placebo) or the same amount of phosphate-buffered saline solution (control) were injected into the upper joint cavities of both joints of another four male guinea-pigs. At 1, 2, 3 and 4 weeks after injection, the joints were dissected and the expression of delivered LacZ was examined by 5-bromo-4 chloro-3-indolyl-beta-D-galactopyranoside (X-gal) staining and reverse transcriptase-polymerase chain reaction (RT-PCR). To investigate the expression of transferred gene in other organs, total RNA was extracted from liver, kidney, heart and brain and the expression of LacZ mRNA and 18 S ribosomal RNA were analysed by RT-PCR. Clear expression of LacZ was observed in the articular surfaces of the temporal tubercle, articular disc and synovium of the temporomandibular joints even 4 weeks after injection in the AxlCALacZ-injected group, while no expression was detected in placebo and control groups. Histological examination confirmed that LacZ activity was clearly detected in a few cell layers of the articular surface tissues, which is much more efficient than in a previously study of the knee joint. In the other organs, expression of the delivered transgene was not observed. Based on these findings, direct gene delivery into the articular surface of the temporomandibular joint using the adenovirus vector is feasible as an effective in vivo method. PMID- 10471155 TI - Inhibition of experimental bone resorption and osteoclast formation and survival by 2-aminoethanesulphonic acid. AB - It is known that bone resorption is mediated by osteoclasts, and lipopolysaccharide (LPS) and inflammatory mediators such as interleukin-1 (IL-1) and prostaglandin E2 (PGE2) induce osteoclast differentiation from haemopoietic cells, 2-aminoethanesulphonic acid, which is known as taurine, is an important nutrient and is added to most synthetic human infant milk formulas. In this study, it was found that 2-aminoethanesulphonic acid inhibits the stimulation of bone resorption mediated by LPS of the periodontopathic microorganism Actinobacillus actinomycetemcomitans Y4 in organ cultures of newborn mouse calvaria. The effect of 2-aminoethanesulphonic acid on the development and survival of osteoclast-like multinucleated cells produced in a mouse bone-marrow culture system was also examined. 2-aminoethanesulphonic acid (100 microg/ml) suppressed the formation of these osteoclast-like cells in the presence of LPS of A. actinomycetemcomitans Y4, IL-1alpha or PGE2 in mouse marrow cultures. On the other hand, 2-aminoethanesulphonic acid did not inhibit 1alpha, 25 dihydroxyvitamin D3-mediated osteoclast differentiation. Although IL-1alpha elongated the survival of the osteoclast-like cells, 2-aminoethanesulphonic acid blocked the supportive effect of IL-1alpha on osteoclast survival. 2 aminoethanesulphonic acid showed no effect on the growth of mouse osteoblasts. Finally, it was found that 2-aminoethanesulphonic acid inhibited alveolar bone resorption in experimental periodontitis in hamsters. These results suggest that 2-aminoethanesulphonic acid is an effective agent in preventing inflammatory bone resorption in periodontal diseases. PMID- 10471156 TI - Tritiated thymidine and bromodeoxyuridine double-labelling studies on growth factors and oral epithelial proliferation in the mouse. AB - Mouse tongue epithelium is characterized by a circadian variation in the number of cells undergoing DNA synthesis. Groups of male BDF1 mice were followed over 48 h and a double-labelling method with tritiated thymidine and bromodeoxyuridine used to determine S-phase labelling indices, together with cell influx to and cell efflux from S, at 4-hourly time points. Control animals exhibited diurnal peaks in labelling index at 03:00 with trough activity 12 h later at 15:00. Cell influx peaked at 23:00 with troughs occurring between 11:00 to 15:00. Peak cell efflux occurred at 07:00 with trough activity at 19:00. Animals injected with epidermal growth factor at 05:00 demonstrated a significant fall in both influx and efflux throughout the 48-h period (P < 0.001), but with preservation of labelling indices, suggesting a slower transit of cells through S-phase, whereas epidermal growth factor injected at 15:00 only produced a significant rise in cell-efflux values. Adrenergic stimulation by intravenous phenylephrine/isoprenaline injection at both 05:00 and 15:00 resulted in a significant rise in cell efflux (P < 0.001), although there was also a rise in labelling index in the 15:00 group (P < 0.001). Animals injected with calmodulin at 05:00 demonstrated a significant reduction in labelling index throughout the 48-h period (P < 0.001), but maintained control values for cell influx and efflux, suggesting faster transit of cells through S. Calmodulin injection at 15:00 produced only a significant reduction in cell influx (P < 0.001). Administration of exogenous growth factors significantly alters the normal rhythmical proliferation of oral epithelial cells in a mouse model. These effects appear to be both growth factor- and time-dependent, and may have both physiological and pathological implications. PMID- 10471157 TI - Effects of forksolin, dibutyryl cAMP and H89 on Ca2+ mobilization in submandibular salivary cells of newborn rats. AB - The effects of substances which affect cAMP or the cAMP-dependent protein kinase (PKA) on the inositol 1,4,5-trisphosphate (IP3) and Ca2+ responses to acetylcholine or thapsigargin were investigated in submandibular gland cells of newborn rats. Exposure to forskolin, dibutyryl cAMP or the PKA inhibitor H89 did not affect the formation of IP3 or the release of Ca2+ from intracellular stores elicited by acetylcholine. However, the thapsigargin-induced Ca2+ release was reduced by dibutyryl cAMP and enhanced by H89 in immature cells. Ca2+ influx activated by acetylcholine and thapsigargin was additive in immature cells but not in mature cells, suggesting the presence of a separate Ca2+ entry pathway in immature cells. Moreover, the acetylcholine-stimulated Ca2+ influx was significantly potentiated by forskolin and dibutyrylcAMP, but not by H89 in immature cells. In contrast, the thapsigargin-activated Ca2+ influx was dramatically enhanced by H89, but not by forskolin and dibutyrylcAMP in these cells. This modulation of Ca2+ mobilization by the test substances is different from that observed in mature submandibular cells in which forskolin, dibutyrylcAMP and H89 affected both IP3 formation and Ca2+ release in response to acetylcholine. Therefore, these results suggest differences in the interaction between the cAMP-PKA and the phosphoinositide-Ca2+ signalling pathways of mature and immature salivary cells. The modulation of Ca2+ influx by the cAMP-PKA pathway in immature cells is likely to play a part in the maturation of salivary cells. PMID- 10471158 TI - Localization of neuronal-constitutive nitric oxide synthase and secretory regulation by nitric oxide in the rat submandibular and sublingual glands. AB - The distribution of neuronal-constitutive nitric oxide synthase (ncNOs)-positive nerve fibres was compared immunohistochemically, and the effect of NOs inhibitor and NO scavenger on the secretory response was compared functionally, in the two glands. Numerous ncNOs-positive fibres were distributed around acini in the submandibular gland but scarcely any around acini in the sublingual gland. Within the submandibular ganglion (parasympathetic), the nerve-cell bodies were strongly positive. Within the superior cervical ganglion (sympathetic), the nerve-cell bodies were negative, although some positive nerve fibres were observed. The secretory responses to the electrical stimulation of the chorda were significantly reduced by the NOs inhibitor, N(G)-nitro-L-arginine methyl ester (L NAME, 10(-9)-10(-3) M) in a dose-dependent manner. The NO scavenger, 2-(4 carboxyphenyl)4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO) also reduced the chorda-evoked secretion (10(-9)-10(-6) M). The submandibular secretions evoked by stimulation of the superior cervical ganglion were not affected by L-NAME or carboxy-PTIO. In the sublingual gland, neither L-NAME nor carboxy-PTIO affected chorda-evoked salivary secretion. The histochemical and functional results both suggest that NO plays an excitatory role in the regulation of parasympathetic nerve-induced salivary secretion in the rat submandibular gland, but not in the sublingual gland. PMID- 10471159 TI - Expression of cartilage-derived retinoic acid-sensitive protein during healing of the rat tooth-extraction socket. AB - Cartilage-derived retinoic acid sensitive protein (CD-RAP) is a recently described, cartilage-specific protein. During early healing of the tooth extraction socket, cells express both chondrogenic and osteogenic cell markers, but no cartilage is formed. Cartilaginous collagen type II protein, a major constituent of hyaline cartilage, has not been detected in the healing socket, although type IX collagen, which coats these fibres, has been detected transiently in early socket healing. This study investigated the spatial and temporal expression of CD-RAP and various osteoblast cell markers, i.e., alkaline phosphatase, osteopontin, osteonectin and osteocalcin, during healing. Immunolocalization of these proteins was determined in the rat tooth socket at 3, 4, 5, 6, 7, 8, 10 and 14 days after extraction. CD-RAP was expressed by preosteoblast cells maximally at 6, 7, and 8 days after extraction. Fully differentiated osteoblasts expressed osteocalcin, a specific osteoblast marker. Preosteoblasts and fibroblasts did not express osteocalcin. On double immunofluorescent staining, some preosteoblasts coexpressed CD-RAP (indicative of chondrogenic differentiation), and either alkaline phosphatase or osteopontin (markers of osteogenic stem-cell maturation). There was no colocalization between osteopontin and osteonectin. CD-RAP was unique amongst the cell markers used in that it was expressed by preosteoblasts, but not by osteoblasts lining the newly formed trabeculae. CD-RAP may have an important role in osteoblast cell differentiation during bone healing. PMID- 10471160 TI - Characterization of dental follicle cells in developing mouse molar. AB - Dental follicle has been implicated as the origin of alveolar bone, cementum and periodontal ligament, but there is no direct evidence of their cellular lineage. The present pilot study was designed to characterize the phenotype of cultured cells obtained from the dental follicle of neonatal mouse molars. Developing mandibular molars from 6-day-old CD-1 mice were subjected to 1% trypsin in Hank's balanced salt solution. After trypsinization, the dental follicle was enucleated from the tooth germ and separated from the associated epithelial root sheath. Pure dental follicle tissue was cultured in alpha-minimal essential medium containing 10% fetal bovine serum and antibiotics. The nature of the cultured follicle cells was determined in situ by immunocytochemical staining for type I and III collagen, fibronectin, and alkaline phosphatase expression. Earlier phenotypic markers for mineralization such as bone sialoprotein and osteopontin were also examined by in situ hybridization of matched molar tissues. The extracellular matrix proteins (such as type I collagen and fibronectin) were moderately expressed cytochemically. However, type III collagen was strongly stained. Gene expression of bone sialoprotein and osteopontin was detected in sections of mouse molars of similar age. The ALPase activity showed moderate to strong intensity in these primary cultured cells and responded to 1,25(OH)2 vitamin D3 treatment. Cytokeratin stains were not noted in these cells. In conclusion, the 6-day-old dental follicle cells exhibit partial characteristics of a mineralized tissue-forming phenotype even though the expression of osteopontin, type I collagen and fibronectin was low at this stage. PMID- 10471161 TI - The immunohistochemical localization of phospholipase Cgamma and the epidermal growth-factor, platelet-derived growth-factor and fibroblast growth-factor receptors in the cells of the rat molar enamel organ during early amelogenesis. AB - Findings on the localization and possible roles of the major growth factors, epidermal (EGF), platelet-derived (PDGF) and fibroblast (FGF) in early amelogenesis are contradictory and inconclusive. This study sought to localize immunohistochemically phospholipase (PLCgamma) and the EGF, PDGF and FGF receptors in the cells of the enamel organ during the events leading directly to early enamel formation in rat molars. PLCgamma is an immediate, downstream, signal-transduction pathway effector unique to the three receptors. A whole-head, freeze-dried sectioning method was used to reduce the possibilities of false negative staining. A modification of the avidin/biotin complex method of immunohistochemical localization was used. Anti-PLCgamma and antibodies to each of EGF, PDGF and FGF receptors colocalized in the preameloblasts of the cervical loop, adjacent to the undifferentiated mesenchymal cells of the dental pulp. This staining disappeared shortly after the beginning of dentine mineralization. Staining for all four antibodies appeared on the proximal ends of the differentiating presecretory ameloblasts at the level of the beginning of predentine matrix deposition and continued in the secretory ameloblasts. It appears that EGF, PDGF and FGF have roles in the differentiation of ameloblasts and in control of cellular functions in presecretory and secretory ameloblasts. Their roles may represent redundancy of the kind seen in highly conserved tissues. PMID- 10471162 TI - Analysis of oestrogen receptor mRNA by reverse transcriptase-polymerase chain reaction in human periodontal ligament cells. AB - Periodontal ligament (PDL) cells have osteoblast-like features and are capable of differentiating into osteogenic cells. As human osteoblasts express oestrogen receptor mRNA, it is possible that PDL cells do so also, but findings have been conflicting. To determine whether they do express oestrogen receptor mRNA, the reverse transcriptase-polymerase chain reaction was performed with two different primers. Cells were obtained from a healthy periodontal ligament of premolar extracted for orthodontic reasons. The human breast adenocarcinoma cell-line MCF7 was used as a positive control. Expression of oestrogen receptor mRNA was detected in PDL cells with one of the primers but with less intensity than in MCF7 cells. Southern hybridization confirmed these results. These findings suggest that PDL cells express oestrogen receptor mRNA at low levels. PMID- 10471163 TI - Stress, coping, and adjustment in children with a chronic disease: a review of the literature. AB - PURPOSE: To review the literature on the consequences of having a chronic disease on the child's functioning in daily life. METHOD: A wide search of the literature resulted in the location of around 200 empirical studies with a focus on chronically ill children. This body of literature is discussed in three parts: (1) the academic and psychosocial adjustment of chronically ill children, (2) the ways chronically ill children cope with disease-related stress and other stressors, and (3) the effectiveness of coping strategies. RESULTS: Children with a chronic disease do not show lower school performance despite higher absence rates (an exception is epilepsy). Their self-concept is similar to that of healthy children. However, they show more behaviour problems, especially internalizing problems such as depression and social withdrawal. The authors could not find evidence for the claim that children with a chronic disease are more frequently confronted with stress than their healthy peers. Children with a chronic disease use a variety of coping strategies to deal with various disease related and common stressors. The coping strategies they use in relation to common stressors appear to be similar to those of healthy children. CONCLUSION: Incidences of maladjustment vary across studies and different chronic diseases. Most studies on coping lack situational sensitivity, which makes it impossible to compare results. Findings on the effectiveness of these children's coping strategies are still scarce and inconclusive. Recommendations for future research on coping with chronic disease in childhood are given. PMID- 10471164 TI - Promoting group empowerment and self-reliance through participatory research: a case study of people with physical disability. AB - PURPOSE: Despite the growing popularity of the empowerment construct among social scientists, relatively few empowerment studies involving groupwork with people with physical disabilities exist. This article accordingly describes and analyses the organic development of the empowerment process within a spinal cord injury self-help group, set against the backdrop of policy imperatives for disability in post-apartheid South Africa. METHOD: The treatise on the group empowerment process is located within the context of a group evaluation conducted within a participatory research framework. Key variables informing the research approach included: quality of participation, control over resources and decision-making, shift in critical consciousness and understanding, malleability of roles within the group and role of the health professional. RESULTS: Group members assumed ownership of group management and decision-making and shifted from a professionally-led to a peer-led self-help group. Group objectives changed from providing mutual support to community education and outreach activities. The role of the health professional shifted from group facilitator to invited consultant. CONCLUSIONS: This case study demonstrates how group participation, promoted by a critically informed therapeutic and research praxis, can unlock the inherent potential for self-reliance and empowerment of socially marginalized collectives. It offers important insights with regard to group process, participatory research and the role of the health professional in creating opportunities for empowerment and self-reliance of people with disability. PMID- 10471165 TI - From handicap to disability: language use and cultural meaning in the United States. AB - The historical shift in discourse regarding disabled persons in the US is analysed by its core terms, handicap and disability. First, this shift in discourse is illustrated in the marketplace and in the context of the Americans with Disabilities Act. Then, discourse shifts are theoretically explicated through a semiotic and historical analysis. Last, the discouse shifts are theoretically explicated through a semiotic and historical analysis. Last, the discourses of handicap and disability are tested in their societal context, especially in the case of American youngsters who have directly experienced the shift in their own life course. Changes in terminology reflect a different logic of the understanding of disability as phenomenon. While 'handicap' reflects an era in which accessibility is central for understanding and practice, the concept of 'disability' shifted the attention to the individual's ability. The impact of such large discourse shifts is tangible in the lives of individuals with mental retardation in the experiences that shift from 'access' to 'ability'. Whereas in the handicap era, access is to be created, in the disability era this is replaced by ability. Professionals are reminded that the rehabilitation process takes on different meanings with shifting discourses. PMID- 10471166 TI - The epidemiology of comorbidity between depression, anxiety disorders and somatic diseases. AB - Depression, generalized anxiety disorder and somatoform disorder are the most common psychiatric disorders in primary care and in the general population. These disorders also show an excess comorbidity which is substantially more common than expected on the basis of their prevalence rates both in primary care as well as in general population. There is no unique mechanism accounting for each of these excess comorbidity constellations. Multiple consequences underline the clinical relevance of comorbidity between these disorders: more psychosocial disability, substantially elevated risk for completed suicides and suicide attempts, worse clinical outcome and treatment response. Recognition of comorbidity is essential for improvement of treatment. PMID- 10471167 TI - Light treatment of seasonal affective disorder in combination with citalopram or placebo with 1-year follow-up. AB - This placebo-controlled, double-blind, 1-year pilot study aimed at investigating possible clinical advantages of combining initial light therapy with the selective serotonin reuptake inhibitor (SSRI) citalopram as well as the effects of continuous long-term administration of this drug in patients with seasonal affective disorder (SAD). Eight physically healthy women who met the DSM-III-R criteria for SAD were included in the study. Four women were randomized to the citalopram group receiving 40 mg citalopram daily from the first of 10 light treatment days and throughout the 1-year study. The remaining four women were allocated to the placebo group using the same double-blind repeated measures design. The clinical outcome was measured by using three versions of the Comprehensive Psychopathological Rating Scale (CPRS) and Visual Analog Scales (VAS), respectively. Taking the initial rating scores into account in covariance analyses, no statistically significant group difference was found during the light treatment period. However, during the follow-up period the full version of the CPRS and the self-rating version of CPRS and the VAS-scales for global condition and depressed mood were statistically significantly lower in the citalopram group compared with the placebo group. Thus, in this small but carefully observed sample of SAD-patients combining initial light therapy and long-term citalopram treatment was clinically more effective over time than the placebo combination. Our findings support the notion that light therapy with concomitant and continued SSRI (citalopram) treatment is a useful strategy to achieve beneficial long-term effects in patients with the SAD syndrome. PMID- 10471168 TI - Antidepressants in panic disorder. AB - Panic disorder is a serious and common illness affecting 1% of the population at any one time. Comorbidity with depression may be as high as 40%. The illness has been recognized as a separate entity since the 1960s and treatment with tricyclics being used since that time. Tricyclics, monoamine oxidase inhibitors, benzodiazepines, beta-blockers and anticonvulsants have all been used with varying degrees of success. Until recently, tricyclics and benzodiazepines were the treatments of choice but selective serotonin reuptake inhibitors (SSRIs) have recently been studied intensively and, based on studies of citalopram and paroxetine, must now be considered first line therapy. Both SSRIs and tricyclics suffer from a long latency period, possibly as long as 12 weeks before maximal benefit is obtained, which is in contrast to the benzodiazepines that produce almost instant symptom relief. The dependency potential of the benzodiazepines, however, limits their usefulness. Paroxetine and citalopram have good efficacy data over both the short and long term and are effective at standard dosages, the most effective for citalopram being 20-30 mg. Both drugs performed better than the comparator tricyclic antidepressant (chlomipramine) and must now be considered current drugs of choice. Despite the superior efficacy of these drugs, however, many patients are poorly controlled and investigation of combination therapies for resistant panic disorder is needed. PMID- 10471169 TI - Beyond depression: citalopram for obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD) and serotonin reuptake inhibitors became inextricably joined with the discovery of their effectiveness for the treatment of OCD in 1969 and 1976. The specificity of response of OCD to serotonin reuptake inhibitors has provided an important platform for the exploration of the neurobiology of OCD. This knowledge has come from many different areas of investigation, including neuroimaging, neuroimmunology and pharmacologic challenge studies. As the research into the pharmacologic treatment of OCD has progressed, a number of serotonin reuptake inhibitors have been approved and marketed for the treatment of OCD. These agents include clomipramine, sertraline, paroxetine, fluoxetine, and fluvoxamine. Because these agents share the same mechanism of action (serotonin reuptake inhibition), choosing which agent to use must be based on other parameters. These include differences in efficacy, dosing, and side-effect profile. Citalopram, a newly marketed selective serotonin reuptake inhibitor could offer some advantages over those agents currently marketed. Theoretically, it should also be effective in the treatment of OCD. In treating depression, it has been shown to have minimal side-effects, low risk of withdrawal symptoms, and little possibility of interaction with other agents. PMID- 10471170 TI - Serotonin reuptake inhibitors for the treatment of premenstrual dysphoria. AB - Premenstrual dysphoria (PMD) is a severe form of premenstrual syndrome, afflicting approximately 5% of all women of fertile age. The cardinal symptoms are irritability and anger. In addition, sadness, tension and carbohydrate craving are common complaints. The symptoms surface regularly between ovulation and menstruation, and disappear completely within a few days after the onset of the bleeding; in patients with remaining symptoms during the follicular phase, alternative diagnoses should be considered. In a large number of recent trials, serotonin reuptake inhibitors (clomipramine, citalopram, fluoxetine, paroxetine, sertraline) have been shown to reduce the symptoms of PMD much more effectively than placebo; in contrast, non-serotonergic antidepressants (maprotiline, bupropion) appear to be ineffective. Interestingly, the onset of action of clomipramine and selective serotonin reuptake inhibitors (SSRIs) is much shorter when used for PMD than when used for depression, panic disorder, or obsessive compulsive disorder. Consequently, patients with PMD can restrict the medication to the luteal phase of the cycle. In a recent placebo-controlled trial, intermittent administration of the SSRI citalopram was shown to reduce the symptoms of PMD significantly better than placebo, but also better than continuous administration of the drug. A reasonable interpretation of the latter, unexpected finding is that continuous medication may be associated with a certain development of tolerance than can be avoided by intermittent drug administration. The observation that the symptoms of PMD may be effectively reduced by SSRIs is of considerable clinical importance since previously no effective treatment for this common condition - apart from those disrupting ovarian cyclicity - has been available. It is also of theoretical importance because it constitutes one of the first pharmacological observations supporting the concept that serotonin may dampen irritability and anger in humans. PMID- 10471171 TI - Metabolic interactions of central nervous system medications and selective serotonin reuptake inhibitors. AB - Selective serotonin reuptake inhibitors (SSRIs) are prescribed alone and in combination with other psychotropic medications in the treatment of a variety of psychiatric disorders. Such combinations create the potential for pharmacokinetic interactions by affecting the activity of the cytochromes P450 (CYP450), drug metabolizing oxidative enzymes. SSRIs are not equivalent in their potential for interactions when combined with other central nervous system (CNS) medication. Generally citalopram and sertraline are characterized by weaker inhibition of CYP450 enzymes and, therefore, hold less potential for interaction than the other SSRIs. Paroxetine potently inhibits CYP2D6, which can result in increased neuroleptic serum concentrations, accompanied by increased CNS side-effects. Similarly, as a potent inhibitor of CYP2D6, fluoxetine can increase serum concentrations of neuroleptics and antidepressants and numerous case reports have documented concomitant adverse events. Fluoxetine also inhibits CYP3A and CYP2C19, increasing serum concentrations of some benzodiazepines. Fluvoxamine is a potent inhibitor of CYP1A2, a moderate inhibitor of CYP3A and a mild inhibitor of CYP2D6. Therefore, interactions with clozapine and benzodiazepines are evident. PMID- 10471172 TI - 1st International Symposium on Adult Immunization in Asia: Prevention of Influenza and Pneumococcal Infections. Hanoi, Socialist Republic of Viet Nam, 20 22 April 1998. Proceedings. PMID- 10471173 TI - The global impact of influenza on morbidity and mortality. PMID- 10471174 TI - The burden of pneumococcal disease among adults in developed and developing countries: what is and is not known. AB - The burden of pneumococcal disease among adults in developed countries is neither widely known nor appreciated. The incidence of pneumococcal pneumonia is uncertain because a precise diagnosis cannot be obtained for most patients. Population-based data on invasive pneumococcal disease (e.g., bacteraemia and meningitis) suggest an annual incidence in all developed countries of > or =15-20 cases per 100,000 persons of all ages and > or =50 cases per 100,000 elderly adults (> or =65 years). In developing countries there are no population-based data on the burden of pneumococcal disease among adults. Studies of high risk groups, hospital-based studies, vaccine efficacy trials, extrapolations from surveillance of "native populations" in developed countries, and demographic studies in developing regions all suggest a high burden of disease. The broad variation in these estimates, however, indicates that better studies are needed. Increased use of pneumococcal vaccines among adults in all countries will depend on better scientific and public understanding of the burden of pneumococcal disease. In developing countries, intensive community-based studies of the impact of pneumococcal disease, or, alternatively, a "vaccine probe" approach, in which a population is vaccinated and the reduction in pneumonia is compared with that in a control population, could give more accurate estimates of the burden of disease and of the potential effectiveness of pneumococcal vaccination among adults. PMID- 10471175 TI - Epidemiological data on influenza in Asian countries. PMID- 10471176 TI - The importance of global surveillance of influenza. PMID- 10471177 TI - Poultry and the influenza H5N1 outbreak in Hong Kong, 1997: abridged chronology and virus isolation. PMID- 10471178 TI - Surveillance and impact of influenza in Europe. Groupe Regional d'Observation de la Grippe and European Influenza Surveillance Scheme. PMID- 10471179 TI - Surveillance and impact of influenza in the United States. PMID- 10471180 TI - Complications of influenza and benefits of vaccination. AB - Uncertainty regarding the benefits of influenza vaccination may contribute to the underutilization of this vaccine. We have conducted serial cohort studies using the administrative data bases of a Twin Cities based managed care organization to assess the impact of disease and benefits of vaccination among the elderly. For the 6 seasons 1990-1991 through 1995-1996, there were more than 20,000 elderly members of the health plan included in each cohort. Data collected included information on baseline demographic and health characteristics, vaccination status and outcomes (hospitalizations and death). Multivariate regression techniques were used to compare the risks of outcomes between vaccinated and unvaccinated persons while controlling for covariates and confounders. Results for data pooled over the 6 seasons demonstrated that influenza vaccination was associated with significant reductions in hospitalizations, outpatient visits, and death among the elderly. Similar findings were observed for low, intermediate, and high risk subgroups. Vaccination was also associated with cost savings. These findings are consistent with results from studies conducted in other countries and over other seasons and strongly support age-based recommendations for annual influenza vaccination for all persons ages 65 and over. PMID- 10471181 TI - Immunization against influenza in the elderly: the Argentinian experience, 1993 1997. PMID- 10471182 TI - Assessment of effectiveness of Vaxigrip. AB - In order to assess the effectiveness of Vaxigrip in 1996-1997, we chose 1356 people in the Chinese Army, who had not recently had influenza and who were divided into an injection group and a control group. The injection group consisted of 80 children aged 3-6 years, 363 adults aged 18-59 years and 235 people over 60 years of age. The control group consisted of 88 children aged 3-6, 372 adults aged 18-59 and 218 people over 60. They were observed from the 21st day to the 6th month after Vaxigrip administration. Each person was requested to report having a body temperature over 38.5 degrees C, headache, myalgia or arthalgia, cough, pharyngodynia or nasal obstruction. Fever due to other causes was not included in the influenza symptoms. We counted the symptoms of influenza and common cold only once during the observation period. Our final finding was that the incidence rates of the influenza and common cold symptoms were reduced by 84.8% in children, 74.0% in adults and 68.6% in elderly people. In comparison with the control group, the incidence rate of influenza-like symptoms was reduced by 71.1% in the injection group, and the incidence rate of common cold symptoms in the injection group was lower than that in the control group. There was no large difference by age. The incidence rate of symptoms of upper respiratory tract infection in the injection group was 47.5% lower than that in the control group. Our findings are basically the same as those in other countries. PMID- 10471183 TI - Immunization against influenza among working adults: the Philippines experience. PMID- 10471184 TI - Vaccination of the older adult: the Australian experience. PMID- 10471185 TI - Clinical effectiveness and cost effectiveness of influenza vaccination among healthy working adults. PMID- 10471186 TI - Epidemiological data on pneumococcal infections in Asian countries. PMID- 10471187 TI - Strategies for the control of pneumococcal diseases. AB - Streptococcus pneumoniae (pneumococcus) is a Gram-positive, encapsulated bacteria that is a major cause of human disease in people of all ages. It is the most important cause of bacterial pneumonia in infancy, childhood and adult life, and the most important cause of meningitis in all age groups except children of 3 months to 2 years in whom Haemophilus influenzae type b (Hib) predominates (in the absence of Hib vaccination). Antibodies to the pneumococcal polysaccharide capsule are protective, and at present 90 capsular serotypes are recognized. The global burden of pneumococcal disease is poorly understood. It is believed to be responsible for 1-2 million deaths among children under 5 years of age every year and probably a similar number among adults. Thus, the global burden of pneumonia in adults is probably significantly underestimated at present. Strategies for the control of pneumococcal disease include control of risk factors, treatment of established cases and vaccination. In children, improved nutrition, better housing and reduced indoor air pollution are difficult to address, but should eventually reduce pneumonia rates. In adults, the risk factors are even more difficult to address, although control of alcohol and tobacco consumption and reduced transmission of HIV should all affect pneumococcal disease rates. Penicillin-resistant pneumococci are now widespread throughout the world. Where penicillin resistance occurs, penicillin should not be used to treat pneumococcal meningitis; however, penicillin, at higher doses if necessary, remains the drug of choice for the treatment of pneumococcal pneumonia, even where penicillin resistance is prevalent. There are three approaches to pneumococcal vaccination: polysaccharide vaccines (covering 23 serotypes), polysaccharide-protein conjugate vaccines (covering 9-11 serotypes) and common protein vaccines (which are not serotype-specific). Only polysaccharide vaccines are available now, but conjugate vaccines will be available soon. Polysaccharide vaccines probably have a role in protecting the elderly from pneumococcal disease, especially those at high risk. The potential role of conjugate vaccines in infants is unclear. PMID- 10471188 TI - The clinical effectiveness of pneumococcal vaccination: a brief review. AB - Randomized controlled trials have shown that pneumococcal polysaccharide vaccine is efficacious in preventing pneumococcal bacteraemia and pneumococcal pneumonia in young adults. Clinical trials in older adults, however, have been inconclusive, usually because the studies have been too small. Retrospective studies have shown that pneumococcal vaccination is approximately 50-80% effective in preventing invasive pneumococcal disease among older persons. Vaccination in this age group is also very cost-effective. These findings are the basis for the recent expansion of immunisation policies and the growth in vaccine use in many developed countries. Serologic and clinical studies, however, suggest that vaccine-induced protection declines after 3-5 years, leading to widespread concern about the need for routine revaccination. Because pneumococcal polysaccharide vaccine does not induce immunologic memory, the benefits of revaccination can also be expected to be relatively short-lasting. Alternative strategies of immunological priming of adults with pneumococcal conjugate vaccine followed by boosting with polysaccharide vaccine, or perhaps vaccination with one of the newer protein vaccines, should be considered. Because these new generation pneumococcal vaccines could provide a foundation of life-long protection against pneumococcal infection, their widespread use among adults could have an immense impact on public health worldwide. PMID- 10471189 TI - The additive benefits of influenza and pneumococcal vaccinations during influenza seasons among elderly persons with chronic lung disease. AB - Uncertainty regarding the benefits of pneumococcal vaccination may contribute to the under use of this vaccine. The present study was conducted to clarify the benefits of influenza and pneumococcal vaccinations during 3 influenza seasons among elderly persons with chronic lung disease. All elderly members of a large managed care organization with a prior diagnosis of chronic lung disease were included in a cohort that was followed over three influenza seasons (1993-1994, 1994-1995, and 1995-1996). Data obtained from the administrative data bases of the health care organization included baseline demographic and health characteristics, influenza vaccination status for each season, date of pneumococcal vaccination, and outcomes for each season including hospitalization for pneumonia and death. Cox proportional hazards regression and Poisson regression with repeated measures were used to compare the risk of outcomes among vaccinated and unvaccinated persons while controlling for covariates and confounders. During the three influenza seasons, influenza vaccination alone was associated with a 52% reduction (95% CI 18-72) in hospitalizations for pneumonia and a 70% reduction (95% CI 57-89) in death. Pneumococcal vaccination alone during the three influenza seasons was associated with a 27% reduction (95% CI 13 52) in hospitalizations for pneumonia and a 34% reduction (95% CI 6-54) in death. Both vaccinations together demonstrated additive benefits. When both vaccinations had been received, there was a 63% reduction (95% CI 29-80) in hospitalizations for pneumonia and an 81% reduction (95% CI 68-88) in death versus when neither had been received. These findings suggest pneumococcal vaccination is associated with substantial benefits for elderly persons with chronic lung disease. PMID- 10471190 TI - Considerations for adding pneumonia and influenza vaccines to public health programmes. PMID- 10471191 TI - Private or national health insurance for adult vaccination in developed countries? PMID- 10471192 TI - Importance of information for influenza control: the French experience. PMID- 10471193 TI - Pneumococcal immunization and public health: the Canadian experience. AB - Pneumococcal vaccine has been poorly used in Canada, despite strong recommendations for its use by the Canadian National Advisory Committee on Immunization. In a recent survey of health officials, however, seven of the 12 Canadian provinces and territories were found to either have a programme for all persons > 65 years of age or were planning to implement one within the next year. Factors that have contributed to this increased interest include: better data on disease incidence and preventable illness from population-based surveillance; data on emerging resistance of Streptococcus pneumoniae in Canada to penicillin and other antimicrobials; implementation of vaccine programmes for the elderly by public health officials in Ontario, Nova Scotia and British Columbia; completion of a cost-benefit study of pneumococcal vaccine for Canada; and increased attention to pneumococcal vaccination at national immunization meetings and in the medical literature. Increased availability of vaccine and competitive pricing are also making programmes for the elderly more feasible and affordable. A national meeting entitled 'Preventing Pneumococcal Disease: A Canadian Consensus Conference' was held in February 1998 to further build on this growing interest. PMID- 10471194 TI - Adult immunization in Asia--summary and recommendations. PMID- 10471195 TI - Influenza surveillance in Australia. PMID- 10471196 TI - Influenza surveillance in New Zealand. PMID- 10471197 TI - Pneumococcal infections in Victoria, Australia. PMID- 10471198 TI - Pneumococcal infections in New Zealand. PMID- 10471199 TI - Circadian rhythms and light responsiveness of mammalian clock gene, Clock and BMAL1, transcripts in the rat retina. AB - Circadian expression and light-responsiveness of the mammalian clock genes, Clock and BMAL1, in the rat retina were examined by in situ hydbribization under constant darkness. A small but significant daily variation was detected in the Clock transcript level, but not in BMAL1. Light increased the Clock and BMAL1 expressions significantly when examined 60 min after exposure. The light-induced gene expression was phase-dependent for Clock and peaked at ZT2, while rather constant throughout the day for BMAL1. These findings suggest that Clock and BMAL1 play different roles in the generation of circadian rhytm in the retina from those in the suprachiasmatic nucleus. Different roles are also suggested between the two genes in the photic signal transduction in the retina. PMID- 10471200 TI - Chronic exposure of rat primary astrocyte cultures to manganese results in increased binding sites for the 'peripheral-type' benzodiazepine receptor ligand 3H-PK 11195. AB - Alterations of 'peripheral-type' benzodiazepine receptors (PTBRs) in brain are a feature of hepatic encephalopathy (HE). Although ammonia toxicity has been implicated in the disorder, recent findings suggest an accumulation of manganese in the brains of cirrhotic patients dying in hepatic coma. In this study, we examined the expression of PTBRs as well as the binding of the selective PTBR ligand 3H-PK 11195 in cultured astrocytes following chronic exposure to manganese. When astrocytes were exposed to 100 microM manganese for 1 week, a 57% increase in Bmax for 3H-PK 11195 binding was detected (P < 0.01) with no change in the Kd value. However, an examination by RT-PCR of the expression of the isoquinoline-binding moiety of the PTBR complex in these cells revealed no change in PTBR mRNA levels following manganese treatment. These findings suggest that manganese up-regulates 3H-PK 11195 binding sites by a process which does not involve a change in transcription. In view of the proposed role of astrocytic PTBRs in 'neurosteroid' synthesis, manganese-induced increases of PTBRs could contribute to the pathogenesis of HE. PMID- 10471201 TI - A comparative study of the beta-adrenoceptors in higher song nuclei of birds. AB - The present study analyses the presence of beta-adrenoceptors in the main telencephalic song nuclei of the goldfinch and parakeet, species belonging to the two most important groups of birds that reproduce learned songs: oscine songbirds and parrots, respectively. Brains of both species sectioned at appropriate levels were used to perform autoradiographic saturation studies using [3H]CGP 12177 as a radioligand. The results show similar K(D)values for both species (0.1-0.3 nM) and striking differences in Bmax. Thus, beta-adrenoceptors are abundant in the telencephalic vocal control nuclei of the parakeet but not of the goldfinch. The predominance of the beta2 subtype in the song nuclei of both species is also confirmed. We conclude that these receptors could be involved in functions unique to the parakeet and therefore may contribute to the greater flexibility of the vocal system of this species. Our findings also support the possible involvement of beta-adrenoceptors in the evolution of the avian brain. PMID- 10471202 TI - Brain water content, glucose transporter densities and glucose utilization after 3 days of water deprivation in the rat. AB - After 3 days of water deprivation, the following parameters were measured in rats: (i) brain water content (apparent diffusion coefficient); (ii) local cerebral glucose utilization (LCGU) ([14C]deoxyglucose method) and (iii) densities of glucose transporters Glut1 and Glut3 (immunoautoradiography). The results show that brain water content is maintained after water deprivation. Densities of glucose transporters Glut1 and Glut3 increased in parallel to increased LCGU in some of the osmoregulatory structures indicating a long-term local adaptation of glucose transporters to LCGU. PMID- 10471203 TI - Morphine applied to the thalamic nucleus submedius produces a naloxone reversible antinociceptive effect in the rat. AB - Our previous studies have indicated that the thalamic nucleus submedius (Sm) is involved in nociceptive modulation and plays an important role in an endogenous analgesic system (a feedback loop) consisting of spinal cord - Sm - ventrolateral orbital cortex (VLO) - periaqueductal gray (PAG) - spinal cord. To further investigate the neurotransmitter and receptor mechanisms in this nociceptive modulatory pathway, we tested the effects of microinjection of morphine and naloxone into the Sm on the rat tail flick (TF) reflex. A unilateral microinjection of morphine (8.0 mM, 0.5 microl) into the Sm significantly depressed the TF reflex, whereas a unilateral microinjection of naloxone (5.0 mM, 0.5 microl) into the Sm facilitated the TF reflex. Five minutes after morphine application into Sm, injection of naloxone in this nucleus markedly reversed the inhibition evoked by applying morphine in Sm. These findings suggest that the endogenous opioid peptides may be involved in the antinociceptive effects evoked by activation of the Sm-VLO-PAG pathway which depressed the nociceptive inputs at the spinal level via the brainstem descending inhibitory system, and exert a tonic descending influence. PMID- 10471204 TI - Kainic acid seizure suppression by neuropeptide Y is not correlated to immediate early gene mRNA levels in rats. AB - Kainic acid induces seizures and a rapid induction of immediate early genes and neuronal death. Neuropeptide Y (NPY) is implicated in seizure inhibiting activity. In order to investigate the mechanisms by which NPY inhibits seizure activity, this study was carried out to measure the levels of mRNAs encoding three different immediate early genes, in regions of the hippocampus and relate their induction to the behaviour in the same animals. NPY inhibited both the time spent in seizures, and the number of generalized seizures. However, NPY did not inhibit the induction of c-fos, FosB or junB mRNA in any hippocampal region examined in the same animals, showing lack of correlation between immediate early gene induction and seizure activity. PMID- 10471205 TI - Trial-to-trial fluctuations in H-reflexes and motor evoked potentials in human wrist flexor. AB - The H-reflexes and the motor potentials (MEPs) evoked by electromagnetic brain stimulation in the human wrist flexor were recorded over many trials. The responses from each stimulus at two steady levels of muscle activation were sorted into three groups, based on their amplitudes. The electromyogram (EMG) in each of these groups was rectified and averaged. The level of pre-response muscle activity was found to correlate with the amplitude of both the averaged H reflexes and the averaged MEPs. This suggests that much of the amplitude fluctuations of both H-reflexes and MEPs can be attributed to moment-to-moment changes in the level of activity of the motoneurone pool. Overall, however, the amplitude of MEPs increased more rapidly than the amplitude of H-reflexes as the pre-stimulus EMG activity increased. This is probably because, while the amplitude of H-reflexes depends primarily on the level of motoneurone pool excitability, the amplitude of an MEP depends not only on this, but also on the excitability of the motor cortex, and the former is to some extent also dependent on the latter. PMID- 10471206 TI - Tactile stimulation of the hand causes bilateral cortical activation: a functional magnetic resonance study in humans. AB - The purpose of the present study was to assess the somatotopy of the cortical sensory representation of the fingers using a natural tactile stimulation of the glabrous skin. Multislice echoplanar imaging techniques were utilized to investigate blood oxygen level dependent (BOLD) signal changes as a measure of cortical activation. Repetitive sensory stimulation of the glabrous skin of digit II-III and digit IV-V resulted in a multifocal signal increase in a restricted area near the central sulcus in the contralateral hemisphere with a considerable overlap between the activated areas of digit II-III and digit IV-V. In addition, in all subjects tactile stimulation resulted in ipsilateral signal increase near the central sulcus, which was 15-22% of the contralateral effect. Stimulation of digit II-III caused significantly (P < 0.05) more activated voxels than digit IV V in the contralateral hemisphere for both hands and for the left hand in the ipsilateral hemisphere. These findings suggest an ipsilateral activation of the primary somatosensory cortex during a natural tactile stimulation of the digits in humans. PMID- 10471207 TI - Cerebrospinal fluid levels of thiamine in patients with Parkinson's disease. AB - Thiamine is an essential cofactor for several important enzymes involved in brain oxidative metabolism, such as the alpha-ketoglutarate dehydrogenase complex (KGDHC), pyruvate-dehydrogenase complex, and transketolase. The activity of KGDHC is decreased in the substantia nigra or patients with Parkinson's disease (PD). We measured cerebrospinal (CSF) levels of thiamine-diphosphate, thiamine monophosphate, free thiamine, and total thiamine, using ion-pair reversed phase high performance liquid chromatography, in 24 PD patients and 40 matched controls. The mean CSF levels of thiamine-derivatives did not differ significantly from those of controls, with the exception of lower CSF free thiamine levels in the PD-patient group. PD patients under levodopa therapy had significantly higher CSF thiaminediphosphate and total thiamine than those not treated with this drug. CSF thiamine levels were not correlated with age, age at onset, duration of the disease, scores of the Unified Parkinson Disease Rating Scale of the Hoehn and Yahr staging in the PD group. These results suggest that low CSF free thiamine levels could be related with the risk for PD. PMID- 10471208 TI - Early changes in nitric oxide synthase activity in atrial intramural arteries following experimental spinal cord injury in rats. AB - Previously, we have shown that immediately after an experimental spinal cord injury (SCI) in anaesthetized rats, there is a large fall in mean arterial pressure (MAP) and heart rate (HR), followed by an abrupt increase in MAP. To evaluate the participation of nitric oxide (NO), we evaluated the activity of nitric oxide synthase (NOS) using Nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry in sections of atria at several post-injury time-intervals. Staining increased at 3 min, reached a maximum at 9 min and diminished 30 min after injury. Pretreatment with atropine prevented changes in MAP, HR and NADPH-d staining suggesting that such modifications result from an increased vagal stimulation. In conclusion, the NOS activity is transiently elevated in the atrial intramural arteries of rats subjected to an SCI. PMID- 10471209 TI - Externally guided control of static grip forces by visual feedback-age and task effects in 3-6-year old children and in adults. AB - This study has been devised to examine the visual feedback control of static grip force levels by pinch and by hand grip during pre-school age and in adults. 69 3 6-year old children and 17 adults were asked to establish and hold grip force levels defined by a visual target and feedback on the dominant and non-dominant hand by hand grip and by pinch grip. From 3 to 6 years of age, the time needed to establish requested grip force levels decreased by a third and the precision increased two-fold for hand grip but four-fold for pinch grip; in contrast to younger children, 5-6-year olds showed a marked superiority of 60% for the pinch grip compared to hand grip, decreasing to about 40% in adults. In the case of pinch grip, all individuals had worse results on higher force levels (children: 50%, adults: 32%). The young children generally tended to use too much force. Older children and adults were better by 75% under the condition of visual feedback vs. internal proprioceptive control (after withdrawing visual feedback). In contrast to previous findings in anticipatory grip force regulation, externally guided force regulation begins to develop during late nursery age. Specific developmental effects were found for grip style and for the ability to use visual feedback and to change from external to internal (proprioceptive) control, and to a lesser extent for force magnitude but not for hand laterality and gender. The findings are interpreted by different developmental velocities of motor areas which are responsible for force regulation mechanisms and for grip style. PMID- 10471210 TI - Cell cycle-related protein expression in vascular dementia and Alzheimer's disease. AB - Recent findings from our and other laboratories indicate that cell cycle-related phenomena may play a key role in the formation of Alzheimer-type pathology and neuronal cell death in both Alzheimer's and cerebro-vascular diseases. In this study we examine the expression patterns of cyclins A, B1, D1 and E in neuronal nuclei in the hippocampus in autopsied healthy elderly individuals, Alzheimer's disease patients and subjects suffering from cerebrovascular disease with and without co-existing Alzheimer's disease. Nuclear cyclin B1 and cyclin E expression was detected in hippocampal neurones in each subject category. However, cyclin B1 expression was significantly elevated in the CA1 of patients suffering from cerebro-vascular disease alone, while cyclin E expression was significantly higher in the CA4 subfield in patients suffering from mixed Alzheimer's and cerebro-vascular diseases compared to subjects in other categories. We hypothesize that cell cycle re-entry may occur in healthy elderly people leading to age-related cell death and mild Alzheimer-type pathology in the hippocampus. However, in pathological conditions, the cell cycle arrest may lead either to the development of severe Alzheimer-related pathology or to excess apoptotic cell death as in vascular dementia. PMID- 10471211 TI - Characterization of spontaneous excitatory synaptic currents in newt retinal bipolar cells. AB - The kinetics of glutamate concentration in the synaptic cleft is an important determinant of synaptic function. To elucidate peak concentration of glutamate released from a single vesicle in the cleft, spontaneous excitatory postsynaptic currents (sEPSCs) in Off-bipolar cells from the sliced newt retina were analyzed using whole-cell patch clamp recording and the computer simulation. The sEPSCs were blocked by an AMPA/kainate (KA) antagonist, 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX), and prolonged by cyclothiazide. However, an N-methyl-D-aspartate (NMDA) antagonist, D-2-amino-5-phosphonopentanoic acid (D-AP5), was ineffective. These suggest that sEPSCs in Off-bipolar cells are mediated exclusively by AMPA/KA receptors. sEPSCs simulated by a detailed kinetic model of AMPA receptor best approximated the data, when peak glutamate concentration was 10 microM. Therefore, it was concluded that peak concentration of glutamate released from a single vesicle would be elevated to approximately 10 microM at the newt Off bipolar dendrite. PMID- 10471212 TI - Cholinergic neurons with monoamine oxidase type B (MAOB)-activity in the laterodorsal tegmental nucleus of the mouse. AB - No neurons in the laterodorsal tegmental nucleus (LDTg) show monoamine oxidase (MAO) activity in the rat or monkey. However, in our recent study, many LDTg neurons with MAO type B (MAOB)-activity were found in MAOA-deficient mice that were derived from C3H mouse line. In the present study, LDTg neurons with MAOB activity were found not only in normal C3H mouse but also in BALB/C and C57BL/6 mouse lines: MAO histochemistry revealed LDTg neurons with MAO-activity even after pharmacological suppression of MAOA-activity with clorgyline, a specific MAOA inhibitor, but not after pharmacological suppression of MAOB-activity with deprenyl, a specific MAOB inhibitor. LDTg neurons with MAOB-activity also showed NADPH-diaphorase-activity, a marker of cholinergic neurons. PMID- 10471213 TI - Lack of effect of the novel anticonvulsant SB-204269 on voltage-dependent currents in neurones cultured from rat hippocampus. AB - The novel anticonvulsant SB-204269 inhibits epileptiform afterdischarges induced by high K+ in rat hippocampal slices. Its effects on voltage-gated Na+ currents, measured from cultured hippocampal neurones using whole cell patch clamp, were compared to the effects of existing anticonvulsants. SB-204269 produced no significant tonic block of Na+ currents nor any voltage-dependent and frequency dependent block at doses 50 to 500 fold higher than its anticonvulsant EC50 of 0.2 microM. In contrast, lamotrigine, phenytoin and carbamazepine at 50 microM, blocked Na+ currents in a voltage-dependent manner. SB-204269 also had no effect on action potential discharges evoked by elevating external K+. These data suggest that direct blockade of voltage-gated channels does not contribute to the anticonvulsant properties of SB-204269 and further support the hypothesis that this compound has a novel mechanism of action. PMID- 10471214 TI - 5-HT4 receptor antagonism potentiates inhibition of intestinal allodynia by 5-HT3 receptor antagonism in conscious rats. AB - Acute levels of distension were applied by balloon to the colo-rectal region in conscious rats and visceromotor responses observed as abdominal muscle contraction; the threshold was typically between 10-40 mmHg. In saline-pretreated rats, the selective 5-HT3 (granisetron) and 5-HT4 (SB-207266) receptor antagonists had no effects on the visceromotor thresholds. 5-Hydroxytryptophan 10 mg/kg, subcutaneously (s.c.) decreased the distension threshold, indicating mechanical allodynia. This increased sensitivity was dose-dependently inhibited by granisetron but was unaffected by SB-207266 100 microg/kg, s.c., a dose which maximally and selectively antagonizes at 5-HT4 receptors. However, this dose of SB-207266 potentiated the inhibitory activity of submaximally-effective doses of granisetron, reducing the ED50 from 0.83 to 0.02 microg/kg, s.c., but without changing the maximum response or the bell-shaped nature of the dose-response curve for granisetron. These data suggest that 5-HT4 receptor activation enhances the ability of 5-HT3 receptor activation to induce intestinal allodynia. PMID- 10471215 TI - C1qB and clusterin mRNA increase in association with neurodegeneration in sporadic amyotrophic lateral sclerosis. AB - We analyzed postmortem tissues of sporadic amyotrophic lateral sclerosis (SALS) for mRNA levels of two inflammatory proteins, complement C1qB and clusterin (apoJ). By Northern blot hybridization, SALS was associated with increased mRNA for C1qB and clusterin in the motor cortex (Brodmann area A4), but not in superior temporal cortex (A17), relative to neurologically normal controls. By in situ hybridization, SALS spinal cords showed increased C1qB and clusterin mRNA in areas undergoing neurodegeneration. This evidence implicates inflammatory mechanisms during neurodegenerative processes in SALS. PMID- 10471217 TI - Remembering the goals of medicine. PMID- 10471216 TI - Advancing the evidence-based healthcare debate. PMID- 10471218 TI - Ideas and ideals in medicine: fruits of reason or props of power? AB - Editors of medical journals play a central role in the promotion -- or suppression -- of ideas and ideals in medicine. Recently eminent among these have been the advocacies of the Evidence-based Medicine (EBM) movement and others concerned with evidence and guidelines for health care. With regard to these topics, it still remains for editors of journals either to advance or to retard even the consolidation of the associated core concepts, most notably those of evidence in medicine, scientific medicine, and rational medicine. I present, first, a case study on the conduct of the editors of three medical journals, specifically their assumption of the role of authority on the scholarly fundamentals of evidence in medicine and their responding to propositions on the topic with commentaries well below the intellectual standards that should prevail in the journals of a learned profession. Then, following a brief review of the Flexnerian and EBM ideas and ideals on the practice of medicine, supplemented by observations drawn from medical sociology and the precepts of the philosophy of science, I posit a way of understanding such behaviour by editors of medical journals. They can have a temptation, and apparently some propensity, to play a regressive role in the development of the fundamentals of medicine. This is prone to occur whenever reason constitutes a threat to power, whether solely to the editors' own or to that of the profession at large. A full realization of the dream of reason in medicine requires an immense integrity of its journal editors and of its other intellectual leaders. PMID- 10471219 TI - Random reflections on science, art and technique applied to medicine and its evaluation. AB - Science is aimed at universal knowledge. Art is aimed at action for solving concrete and local problems. Can medical practice therefore ever be a science even when employing important aspects of scientific practice such as precise concepts and vocabulary, and a demand for well-validated facts together with rational reasoning? Evaluation of medical and clinical practices in general has an ambiguous status. According to the domain to be evaluated, it can reach a level of science in providing results of universal value but more often it is closer to art, the results remaining of local interest. There is no strict correlation between the universal-local axis and the practical interest for public health. Epidemiologists should consider these matters very seriously and aim to contribute to, rather than discard, their significance for the advancement of clinical practice. PMID- 10471220 TI - Reductionism in medicine: some thoughts on medical education from the clinical front line. AB - In clinical medicine, a balance needs to be struck between reductionism (the view that a system can be fully understood in terms of its isolated parts) and holistic complexity appreciation (the view that the whole may not be discernible from knowledge of its parts). Reductionism, which currently is associated with renaming of old well-established ideas, minimizes the reality of complexity in patients with several diagnoses. Holistic complexity appreciation on the other hand may encourage unscientific assessments. A balance between the two needs to be maintained if the needs of students and patients are to coincide. PMID- 10471221 TI - Evidence-based medicine and the real world: understanding the controversy. AB - Controversy has surrounded the 'paradigm' of evidence-based medicine since its introduction in 1992 as a new approach to the teaching and practice of medicine. Here, we address two questions: (1) is evidence-based medicine a good thing?; and (2) why has so much controversy arisen? In addressing these questions, we propose that the discussion surrounding evidence-based medicine should no longer be about whether the application of evidence in clinical practice is a good thing, because it obviously is. Instead, the debate ought to focus on the more difficult question of how to enhance its acceptability among busy clinicians practising in the 'real world'. For the future, we optimistically anticipate an enhanced adoption of evidence-based medicine, as clinicians will become increasingly capable of efficiently accessing existing and forthcoming evidence resources. PMID- 10471222 TI - Examining the assumptions of evidence-based medicine. AB - Evidence-based medicine (EBM) has had enormous impact on clinical medicine, attracting both converts and detractors. The basic premise, that clinical decisions should be made on evidence which is as sound as possible, appears unassailable. Why, then, has the idea stimulated such controversy? In this article, I examine six assumptions of EBM, and point out how these assumptions are frequently open to challenge. Finally, I make specific recommendations for future activities to ensure that the strategy achieves an appropriate role in health care decision-making. These include: (1) conducting research to establish the effectiveness of EBM; (2) incorporating (as they apparently are) more holistic perspectives on the nature of evidence; (3) moving (as they are) toward the use of expert reviews and away from a focus on the skilled individual practitioner; (4) examining the literature on continuing education to devise more appropriate dissemination strategies, including, specifically, educational influentials; and (5) making specific attempts at conciliation and seeking common ground with other schools of thought. PMID- 10471223 TI - Evidence-based psychiatric practice: doctrine or trap? AB - Medical practice has rapidly shifted towards an 'evidence-based' approach. While there are acknowledged clear benefits to this, a number of pitfalls are frequently not appreciated. Perhaps the most important limitation is the extent to which the current body of data is inadequate for many common clinical decisions. Algorithms risk being developed, frequently by third parties, without acknowledgement of these limitations and with substantial implications for clinical independence and the quality of patient care. This paper discusses potential problems of the evidence-based approach and suggests possible guidelines for the management of clinical decisions given the limitations of data based guidelines. PMID- 10471224 TI - Practice standards: the need for a personal element. AB - With rare exceptions, guidelines for clinical practice lack a personal element that allows for physicians' varying goals and patients' differing preferences. The importance of this element is illustrated by means of four examples: 1, hormone replacement therapy for the menopause; 2, early detection of breast cancer; 3, the treatment of acute streptococcal pharyngitis and 4, the diagnosis of symptoms that do not implicate any specific organ system. The advantages and disadvantages of guidelines are pointed out. Among the advantages are their usefulness as standards for audit and the fact that, in the process of reaching consensus, personal experience is taken into consideration. Chief among the disadvantages is their possible misuse by people outside of the profession. It is concluded that guidelines can be improved if their recommendations consider individual goals and preferences. They can become more influential if attention is paid to their proper dissemination. PMID- 10471225 TI - On the scientific inference from clinical trials. AB - We have not been able to describe clearly how we generalize findings from a study to our own 'everyday patients'. This difficulty is not surprising, since generalization deals with how empirical observations are related to the growth of scientific knowledge, which is a major philosophical problem. An argument, sometimes used to discard evidence from a trial, is that the patient sample was too selected and therefore not 'representative' enough for the results to be meaningful for generalization. In this paper, we discuss issues of representativeness and generalizability. Other authors have shown that generalization cannot only depend on statistical inference. Then, how do randomized clinical trials contribute to the growth of knowledge? We discuss three aspects of the randomized clinical trial (Mant 1999), First, the trial is an empirical experiment set up to study the intervention on the question as specifically and as much in isolation from other -- biasing and confounding -- factors as possible (Rothman & Greenland 1998). Second, the trial is set up to challenge our prevailing hypotheses (or prejudices) and the trial is above all a help in error elimination (Popper 1992). Third, we need to learn to see new, unexpected and thought-provoking patterns in the data from a trial. Point one -- and partly point two -- refers to the paradigm of the controlled experiment in scientific method. How much a study contributes to our knowledge, with respect to points two and three, relates to its originality. In none of these respects is the representativeness of the patients, or the clinical situations, crucial for judging the study and its possible inferences. However, we also discuss that the biological domain of disease that was studied in a particular trial has to be taken into account. Thus, the inference drawn from a clinical study is not only a question of statistical generalization, but must include a jump from the world of experiences into the world of reason, assessment and theoretical judgement. PMID- 10471226 TI - Implementing and sustaining evidence-based clinical practice in Australia: the Evidence Based Clinical Practice Research Initiative. AB - Evidence-based medicine is a process by which clinicians translate clinical information needs into answerable questions, track down answers to those needs as efficiently and effectively as possible using the best evidence available, apply the information to patients and evaluate their performance. Implementing evidence based medicine is fragmented and variable. The National Health and Medical Research Council of Australia has implemented a research program to determine successful strategies for implementation and sustaining of evidence-based clinical practice. In addition this research program will help to answer whether evidence-based practice improves patient outcomes. This paper describes this program and other strategies that improve the application of evidence-based practice in clinical settings. PMID- 10471227 TI - A critical evaluation of clinical practice guidelines in neonatal medicine: does their use improve quality and lower costs? AB - Clinical practice guidelines and care pathways have become a focus of quality improvement efforts in Neonatology. Health care administrators believe that using clinical practice parameters reduces health care costs, improves quality of care, and limits malpractice liability. Practice guidelines and surveys of consumer satisfaction have grown in use partly because third-party payers, insurers, and health maintenance organizations, as well as hospital administrators bent on reducing variable costs of care and contracting for capitated care have championed their development, implementation, and monitoring. Overall there is minimal evidence-based medicine to support that neonatal outcomes have benefitted from their implementation, although some studies show affirmative effects in limited populations or in a limited number of centres. For highly autonomous physicians and nurses this standardization of medical decision making may represent a difficult transition into efforts to improve quality, based on evidence-based care, and in some instances into corporate medicine. By realigning the traditional values of patient relationships, including parent involvement, the implementation of guidelines has been fast-tracked in some institutions, without appropriate audit to determine their effectiveness in achieving their goals. However, because guidelines and clinical pathways are here to stay, neonatologists need to think critically about how their content and method of implementation, monitoring and modification may influence medical and nursing teaching and decision making in the future. If guidelines are introduced primarily as a cost savings or containment tool that ignores their impact on the quality of medical care and thereby restricts needed care, then neonatologists must be quick to challenge the potentially damaging and inappropriate use of guidelines and care pathways. Several international efforts are underway to study both the impact of evidence-based guidelines and to determine if they can be imported from one care system into another. Furthermore, there are many medico legal implications of guideline and clinical pathway implementation that may not favour physicians and other neonatal care practitioners, and leave to hospitals, insurers, and ultimately the courts, decisions regarding evidence-based care. Neonatologists and other practitioners in neonatal care centres should critically analyse the goals of guideline development, implementation and monitoring and should restrict themselves to guideline directed care only at those practices for which there is evidence supporting their implementation and continuous monitoring. PMID- 10471228 TI - The Clinical Oncology Information Network (COIN) Project: background, purpose and products. AB - The Clinical Oncology Information Network (COIN) Project of The Royal College of Radiologists is developing evidence-based practice guidelines, clinical core data sets to audit compliance and a dedicated clinical workstation with a client server architecture that will collect the data sets as an automatic by-product of the routine delivery of care. Guidelines for the treatment of breast, colorectal, lung and prostate cancer and for the delivery of chemotherapy and radiotherapy will be published in 1999. Version 2 of a demonstrator workstation is in use at the North Middlesex Hospital, London. PMID- 10471229 TI - A method for guideline development: assessing practical feasibility and adaptation of thyroid nodule guidelines. AB - The objective of this study was to assess adaptation and feasibility of practice guidelines for thyroid nodule management. For one month physicians completed a self-administered semi-structured questionnaire concerning a draft version of the guidelines for each consecutive patient. A sample group of 20 endocrinologists, 20 surgeons and 120 general practitioners from all parts of France took part in this study. The main outcome measures were whether the case description was found by the physicians, and whether recommendations were found and agreement with these recommendations were assessed. We found that two hundred and fifty-three patients (of whom 85% were women) were included in the study: 52% were seen for a surgical decision; 20% for diagnosis or screening and 28% for follow-up. Four percent of the women were pregnant; 2% of the patients had a large nodule; and 2%, a retrosternal nodule. These last three situations had not been well identified in the draft guidelines. The patient's case description was appropriately identified in 75% of the responses. Most cases reported as being not found were, however, covered by the guidelines. Physicians in private practice were less likely than the others to identify their patient's cases. Physicians agreed with the recommendation in 77% of cases. General practitioners were more likely and endocrinologists less likely than surgeons to disagree. Most disagreements and alternative proposals were not evidence-based. Our conclusion is that this study shows that the draft guidelines were applicable, and allowed minor improvements of the final version. Adapting guidelines to practice could improve their use and, consequently, the quality of health care. PMID- 10471230 TI - Using the Internet to teach melanoma management guidelines to primary care physicians. AB - The aim of this study was to determine whether a brief Internet-based education programme could improve physicians' abilities to manage pigmented skin lesions. A pre-test-post-test assessment was used of subjects' knowledge of skin cancer, confidence in their management abilities and actual ability to recommend appropriate treatment for 20 hypothetical patients with pigmented skin lesions. The setting was the general medicine service of an academic medical centre. Seventeen volunteer medical students, house officers and faculty members took part in the study. Following the pre-test, subjects completed a 1-hour computer based educational programme, distributed via the Internet, presenting a guideline for recognizing and managing potentially malignant pigmented skin lesions. The guideline was based on the ABCD rule and the Glasgow seven-point checklist. The educational programme had a positive effect on the subjects' overall skin cancer knowledge and had significantly positive effects on their confidence and ability to apply the management guideline. Based on the guideline criteria, the subjects made the correct management decision on the clinical scenarios 63.2% of the time before the programme and 74.1% of the time after the programme (P = 0.002). We were able to teach melanoma management guidelines to physicians and medical students using a brief, interactive computer programme distributed via the Internet. Such an approach is more cost-effective than classroom teaching and could be used to improve the clinical skills of practising physicians to recognize and manage early melanomas. This approach to distributed learning could also be used to teach other clinical guidelines to physicians. PMID- 10471231 TI - Compliance with US asthma management guidelines and specialty care: a regional variation or national concern? AB - The objective of this study was to examine the compliance with the National Asthma Education Program (NAEP) guidelines among asthmatic members of eight health plans (regions) in seven states, as well as the factors related to the compliance. Information was gathered by means of a cross-sectional survey in a managed care environment. The participants were 6703 respondents (ages 14-65) with moderate or severe asthma. The main outcome measures were compliance with the NAEP guidelines on the use of inhaled steroids, inhaled beta2-agonists, peak flow measurement, and allergy evaluations. Among the results of this survey we found that although these health plans are located from the West Coast to the East Coast and the socioeconomic status of their members varied greatly, compliance with the NAEP guidelines was low among asthmatic members across all geographical regions. The major areas of low compliance identified were inappropriate pharmacological therapy, lack of objective measurement of lung function through peak flow meter, and insufficient environmental trigger control. The regression analyses indicated that the effect of the health plan explained little of the variation in compliance across these regions (only 0.3% at maximum). Low compliance was associated with young age, smoking, moderate asthma, being asthmatic for a few years, currently working, and being treated by a generalist rather than a specialist. In conclusion, this study showed that the compliance with the national guidelines for asthma care was consistently low across different geographical regions in the nation. Improvement in care for asthmatics will require greater commitment and involvement by all stakeholders including physicians, patients, health plans, and employers. We suggest a need for a national strategy to disseminate clinical guidelines not only to the medical community but also to patients themselves. PMID- 10471232 TI - Implementing best practice guidelines: the influence of personal characteristics. AB - Clinical practice guidelines (CPGs) have not been widely adopted by general practitioners despite their obvious benefits of improving health care. Personal characteristics have been identified as one factor influencing doctors' attitudes towards guidelines. This study examined the impact of personal characteristics of Australian general practitioners on their attitudes towards guidelines. Favouring a fee-for-service remuneration system is highly associated with a negative view towards guidelines. This finding needs to be taken into account when developing strategies for the implementation of guidelines in the Australian context. PMID- 10471233 TI - Health policy directions for evidence-based decision making in Canada. AB - The topic, evidence-based decision making, may be seen from many perspectives and levels of policy consideration. Canada has been actively pursuing policy macro level directions for health information and evidence-based decision making through the National Forum on Health and the Advisory Council on Health Infostructure, together with changes in the federal health ministry, Health Canada, and other important national health organizations such as the Canadian Institute for Health Information. It remains to be determined how these initiatives will fundamentally influence a culture of evidence-based decision making and health information availability and quality for the average Canadian. Canada has, nonetheless, recognized that our health care delivery system has fallen behind in terms of using information management and technology to enable and facilitate the ever-growing information needs of the system. Policy direction provided by the National Forum on Health and the Advisory Council on Health Information offer an agenda for action and strategic direction drawn from two broadly based, highly iterative processes. PMID- 10471234 TI - The role of research evidence in pharmaceutical policy making: evidence when necessary but not necessarily evidence. AB - The use of research evidence in policy making at the legislative and administrative levels would appear to be very selective. Focusing on pharmaceutical policy, this paper argues that research evidence is only one ingredient leading to a policy decision and that any examination of research transfer into policy must take into account the many other factors which impact on decision making. The paper describes the policy making process, barriers to the uptake of research evidence into policy and ways of improving research uptake into policy making. Examples are given from drug licensing, remuneration policies, post-marketing surveillance and product withdrawal from the market. PMID- 10471235 TI - Clinical research methods for the new millenium. PMID- 10471236 TI - Health informatics in the Asia Pacific region. AB - Although health informatics has been an established science in some Asia Pacific countries since the 1970s, its current degree of awareness among countries in this region can largely be attributed to the efforts of the Asia Pacific Association for Medical Informatics. These efforts have included the spawning of national health informatics associations in some countries, creating opportunities for cross-country scientific interactions at national health informatics meetings and promoting regional health informatics activities and expertise through the APAMI web site. This presentation gives a review of the current status of health informatics activities among APAMI member countries as well as a cross-section of some of their health informatics projects. PMID- 10471237 TI - Information technology in primary health care. AB - Appropriate application of information technology in primary health care will extend traditional diagnosis and patient management beyond the doctor's clinic into the everyday living environment. We describe a model of information management in primary health care, and place special emphasis on the emerging areas of clinical decision support, computerised clinical measurements, patient education and network connectivity. Briefly discussed is the design of innovative home monitoring techniques and a telemedicine based communications infrastructure that should improve access to high quality primary health care for all citizens, irrespective of their distance from major urban centres. A preliminary design for a telemedicine-assisted primary health care network is presented, based on this model of information management. The premise is that improvements in health care services and reductions in health care costs can be effected by establishing a continuum of patient care from the patient's home, to the doctor's surgery, to speciality services in hospitals and to other service providers in the health care sector. While, the proposal focuses on new opportunities arising from the imminent introduction of broad band interactive fibre optic networks throughout Australia, the technology and projected data transfers could easily be handled in the short-term using modem access to the standard telephone network. A simple connectivity scheme for system integration is also presented. PMID- 10471238 TI - High quality image oriented telemedicine with multimedia technology. AB - Researchers at Osaka and Kyoto University hospital performed three experiments, beginning in 1995, which looked at high quality-oriented telemedicine. This paper describes the system design for the three projects. Experiment 1 applied high definition TV images and B-ISDN for distance learning and medical information exchange. Experiment 2 developed a super high-definition medical image filing system and the images were transmitted via B-ISDN for teleconferences and experiment 3 utilized digital, high-definition, TV images and communication satellites for teleconferences. Multimedia and communication technologies were considered to be fundamental components of telemedicine. The three projects were evaluated initially for quality of images, operability and utility. The experimental design and its implementation showed that it was possible to provide high quality image-oriented telemedicine in the health care environment. Obstacles to establishing practical telemedicine are also discussed. PMID- 10471239 TI - Lessons from a failed information systems initiative: issues for complex organisations. AB - Organisational issues are now recognised as key factors in the effective use of information technology (IT). This paper extends our knowledge of these issues for large complex distributed organisations. It is based on an analysis of the failure of a large IT system selected by a major state public health service for Statewide implementation. The system selected was well proven overseas, was implemented through a pilot process, yet was withdrawn because it did not meet requirements. The issues which are dealt with include the nature of the organisation and its interaction with the implementation strategy, the nature of decision-making, the contrast between the incentives and perspectives at different levels, ownership of the program, management of skills and the impact of instability and complexity. The option of decentralised IT management is raised and the advantages of common systems discussed. A proposal is ventured for managing such developments while maintaining essential commonality. The paper demonstrates that the environment of large complex distributed organisations introduces a complex of issues that are not normally considered in implementation strategy. PMID- 10471240 TI - The application of a computerized problem-oriented medical record system and its impact on patient care. AB - The present computer system is the first of its kind based on problem-oriented medical record (POMR) design developed and operated in a hospital in Hong Kong. It went live in May 1996 with two workstations installed in the medical record office (MRO). Doctors have no direct access to it. They dictate medical notes on tape using either structured or free dictation format, and the tape is brought to the MRO for processing. The principal aim of this study is to study the impact of the computer system on patient care. Retrospective review of medical records and in-depth interviews were conducted to study the quality of medical records and doctor's opinions. A total of 400 manual and 398 computerized patient records were randomly selected for review. The completeness of the manual notes and computerized notes using free dictation format were about the same. The computerized records using structured dictation format may be more complete than those using free dictation format. The in-depth interview shows that most doctors preferred structured medical records but some disagreed with too detailed a level of structuring. They were not familiar with POMR, and some even thought that breaking down the record by problem was not possible. All felt that the present system would not directly affect patient care, but some said that it would facilitate research. In conclusion, since the utility of the information mainly depends on the doctors' efforts, commitment to the agreed structure and subsequent routine audit of computerized medical records are essential to make sure that diagnoses are accurately coded and information is correctly structured. PMID- 10471241 TI - Detecting adverse drug reactions to improve patient outcomes. AB - Adverse drug reactions and inappropriate administration of medications account for poor outcomes for patients. They place patients in life-threatening situations, lead to increased health care costs, extend length of stay in hospitals, as well as increasing litigation. This paper will highlight the incidence of adverse drug events (ADE) in health care and show the low rate of detection within conventional medical records. I will also show how electronic medical records (EMR) improve detection of ADE, enhance clinician compliance to their management, improve patient outcomes, and reduce health care costs. PMID- 10471242 TI - The General Practice computer system project: a doctor's desktop for Australia. AB - The Australian Department of Health and Family Services engaged the IBM consulting practice to develop a functional specification and technical architecture for a General Practice computer system (GPCS) in January, 1997. The project was completed in September, 1997. This paper describes the rationale for development of the specification, the process that was undertaken and provides an overview of the completed specification and architecture. The paper also explores a number of issues related to computing in general practice which were raised during the consultative process, and considers factors which were found to be important in obtaining adoption and use of computer technology on the doctor's desk. PMID- 10471243 TI - The Internal Medicine Center--an integrated solution for information on internal medicine on the World Wide Web. AB - The World Wide Web (WWW) has grown from being a resource center for a select group of scientists to a large database of information that is available to both professionals and public. The amount of medical information on the Web has been increasing exponentially and thus, it has become increasingly difficult for anyone to be able to search for a specific quantum of information among this mass. Even in mid 1996, it was noted that the amount of information on internal medicine was growing rapidly. Hence, on 21 March 1997, the Internal Medicine Center (IMC) was created and launched. The IMC is an unique concept and it represents the first time that medical data on the web has been organized into a form that intimately parallels clinical medicine. The rationale behind the creation of the IMC can be summarized in three words: information, speed and convenience. The interface used by this center reflects these goals because overuse of large image files are avoided, hence decreasing the access time and yet keeping the information in an easily comprehensible manner. In conclusion, the IMC serves as a useful tool for the layman as well as the expert because the comprehensive information that it offers can be accessed rapidly and conveniently. PMID- 10471244 TI - The future of health informatics. AB - Whatever a future vision for health informatics entails, it must take into account the evolving nature of the field, a growing trend towards primary and preventive care and the explosive growth in global networking as exemplified by the Internet. While, historically, storage and retrieval of data has been the main target for information systems development, the need to capture knowledge itself is becoming the focus for development. In parallel, education in health informatics for tomorrow's healthcare professionals is now essential. The Asia Pacific Association for Medical Informatics (APAMI) is a regional group of the International Medical Informatics Association (IMIA). While the newest of the IMIA regional organizations, its growth and activities in the Asia Pacific region aim to advance health informatics. Its triennial conferences act as a means of promoting and monitoring the growth of our field in this region, APAMI itself is a part of the future of health informatics. PMID- 10471245 TI - Selective serotonin-reuptake inhibitors: an update. AB - Selective serotonin-reuptake inhibitors (SSRIs), including fluoxetine, sertraline, paroxetine, fluvoxamine, and citalopram, represent an important advance in the pharmacotherapy of mood and other disorders. They are chemically unrelated to tricyclic, heterocyclic, and other first-generation antidepressants. SSRIs are the treatment of choice for many indications, including major depression, dysthymia, panic disorder, obsessive-compulsive disorder, eating disorders, and premenstrual dysphoric disorder, because of their efficacy, good side-effect profile, tolerability, and safety in overdose, as well as patient compliance. A review of the literature was conducted using Medline and the terms "SSRIs," "fluoxetine," "sertraline," "paroxetine," "fluvoxamine," and "citalopram." Articles were limited to those published in English within the last 15 years. The search revealed that indications for antidepressants include unipolar depression, dysthymia, bipolar depression, treatment-resistant depression, depression in the medically ill, panic disorder, obsessive-compulsive disorder, eating disorders, social phobia, and premenstrual dysphoric disorder. One SSRI, fluoxetine, has demonstrated safety in pregnancy. Side effects of SSRIs include gastrointestinal disturbances, headache, sedation, insomnia, activation, weight gain, impaired memory, excessive perspiration, paresthesia, and sexual dysfunction. PMID- 10471246 TI - Treating depression in patients with cardiovascular disease. AB - Cardiovascular disease and depression are intimately related illnesses. Cardiovascular mortality is more common in persons with depression, and depression following a myocardial infarction is associated with significantly poorer cardiac outcome. Safe and effective simultaneous treatment of depression and cardiovascular illness can be difficult because of the interplay between these conditions. We examine the evidence for cardiovascular effects of depression, as well as the proposed mechanism for these effects. We also review the cardiovascular effects of antidepressant treatments and the mood-altering effects of common cardiovascular medications. Articles reviewed were derived from a Medline search of English-language articles published between 1970 and 1998 (search terms: cardiovascular disease, antidepressants, psychiatry, myocardial infarction, antihypertensive agents, depression). PMID- 10471247 TI - Misdiagnosis of Munchausen syndrome by proxy: a literature review and four new cases. AB - The term "Munchausen's syndrome by proxy" (MSBP) was first used in the 1970s to describe a potentially lethal variant of abuse. In representative cases a mother deceives physicians into treating her child for illness that she has fabricated or induced, her motivation being to accrue the intangible benefits of the "sick role." Increased efforts to identify and protect victims have sometimes resulted in misdiagnosis of MSBP, leading authorities to remove children from the home and/or bring criminal charges against an innocent parent. The authors believe that Improved child protection can be achieved by examining "what went wrong" in cases of erroneously diagnosed MSBP. A review of more than 200 articles and books yielded seven reports of misdiagnosed MSBP in the medical/surgical, psychiatric, nursing, social work, legal, and lay literature. These isolated reports are discussed, and four new cases are presented. The paper concludes with a discussion of lessons to be learned and a table showing situations in which the risk of an erroneous diagnosis of MSBP is increased. PMID- 10471248 TI - To change the patient or the patient's world: the suicide attempt of a teased 12 year-old girl. PMID- 10471249 TI - Small numbers, big impact. PMID- 10471250 TI - Evaluating and improving the appropriateness of treatment for schizophrenia. PMID- 10471252 TI - L'Association pour la recherche sur le cancer. PMID- 10471251 TI - Functional magnetic resonance imaging of personality switches in a woman with dissociative identity disorder. PMID- 10471253 TI - Severe OHSS: An 'epidemic' caused by doctors. PMID- 10471255 TI - P-238 the impact of 6 months GnRH agonist therapy for the treatment of endometriosis on bone marrow density: is there an age related effect? PMID- 10471254 TI - The effect of a difficult embryo transfer on the outcome of IVF. PMID- 10471259 TI - Fourier analysis of the intra-aortic balloon pump PMID- 10471260 TI - Luteovirus interactions with aphid vector cellular components. PMID- 10471261 TI - Congress examines childhood vaccine safety. PMID- 10471262 TI - Apathy in geriatric cancer research. PMID- 10471263 TI - Indecision over human pesticide data. PMID- 10471264 TI - Australia harmonizes bioethics guidelines. PMID- 10471265 TI - Cancer center combines public and private groups. PMID- 10471268 TI - Judging Science: Scientific Knowledge and the Federal Courts. PMID- 10471266 TI - British insurers continue with genetic tests. PMID- 10471269 TI - Rock around the clock. PMID- 10471270 TI - Correction to "Modulation of oncogenic potential by alternative gene use in human prostate cancer" PMID- 10471271 TI - Are presenilins intramembrane-cleaving proteases? Implications for the molecular mechanism of Alzheimer's disease. AB - The amyloid-beta protein (Abeta) is strongly implicated in the pathogenesis of Alzheimer's disease. The final step in the production of Abeta from the amyloid precursor protein (APP) is proteolysis by the unidentified gamma-secretases. This cleavage event is unusual in that it apparently occurs within the transmembrane region of the substrate. Studies with substrate-based inhibitors together with molecular modeling and mutagenesis of the gamma-secretase cleavage site of APP suggest that gamma-secretases are aspartyl proteases that catalyze a novel intramembranous proteolysis. This proteolysis requires the presenilins, proteins with eight transmembrane domains that are mutated in most cases of autosomal dominant familial Alzheimer's disease. Two conserved transmembrane aspartates in presenilins are essential for gamma-secretase activity, suggesting that presenilins themselves are gamma-secretases. Moreover, presenilins also mediate the apparently intramembranous cleavage of the Notch receptor, an event critical for Notch signaling and embryonic development. Thus, if presenilins are gamma secretases, then they are also likely the proteases that cleave Notch within its transmembrane domain. Another protease, S2P, involved in the processing of the sterol regulatory element binding protein, is also a multipass integral membrane protein which cleaves within or very close to the transmembrane region of its substrate. Thus, presenilins and S2P appear to be members of a new type of polytopic protease with an intramembranous active site. PMID- 10471272 TI - Lysine 183 is the general base in the 6-phosphogluconate dehydrogenase-catalyzed reaction. AB - Site-directed mutagenesis was used to change K183 of sheep liver 6 phosphogluconate dehydrogenase to A, E, H, C, Q, R, and M to probe its possible role as a general base catalyst. Each of the mutant proteins was characterized with respect to its kinetic parameters at pH 7 and the pH dependence of kinetic parameters for the K183R mutant enzyme. The only mutant enzyme that gives a significant amount of catalysis is the K183R mutant, and the extent of catalysis is decreased by about 3 orders of magnitude; the general base pK is perturbed to a pH value of >9. All other mutant enzymes exhibit rates that are decreased by about 4 orders of magnitude compared to that of the wild-type enzyme. Data are consistent with the general base function of K183. PMID- 10471273 TI - Farnesyl protein transferase: identification of K164 alpha and Y300 beta as catalytic residues by mutagenesis and kinetic studies. AB - Farnesyl protein transferase (FPT) is an alpha/beta heterodimeric zinc enzyme that catalyzes posttranslational farnesylation of many key cellular regulatory proteins, including oncogenic Ras. On the basis of the recently reported crystal structure of FPT complexed with a CVIM peptide and alpha hydroxyfarnesylphosphonic acid, site-directed mutagenesis of the FPT active site was performed so key residues that are responsible for substrate binding and catalysis could be identified. Eight single mutants, including K164N alpha, Y166F alpha, Y166A alpha, Y200F alpha, H201A alpha, H248A beta, Y300F beta, and Y361F beta, and a double mutant, H248A beta/Y300F beta, were prepared. Steady-state kinetic analysis along with structural evidence indicated that residues Y200 alpha, H201 alpha, H248 beta, and Y361 beta are mainly involved in substrate binding. In addition, biochemical results confirm structural observations which show that residue Y166 alpha plays a key role in stabilizing the active site conformation of several FPT residues through cation-pi interactions. Two mutants, K164N alpha and Y300F beta, have moderately decreased catalytic constants (kcat). Pre-steady-state kinetic analysis of these mutants from rapid quench experiments showed that the chemical step rate constant was reduced by 41- and 30-fold, respectively. The product-releasing rate for each dropped approximately 10-fold. In pH-dependent kinetic studies, Y300F beta was observed to have both acidic and basic pKa values shifted 1 log unit from those of the wild-type enzyme, consistent with a possible role for Y300 beta as an acid-base catalyst. K164N alpha had a pKa shift from 6.0 to 5.3, which suggests it may function as a general acid. On the basis of these results along with structural evidence, a possible FPT reaction mechanism is proposed with both Y300 beta and K164 alpha playing key catalytic roles in enhancing the reactivity of the farnesyl diphosphate leaving group. PMID- 10471274 TI - Conformational states of the nuclear GTP-binding protein Ran and its complexes with the exchange factor RCC1 and the effector protein RanBP1. AB - It has been shown before by (31)P NMR that Ras bound to the nonhydrolyzable GTP analogue guanosine 5'-O-(beta, gamma-imidotriphosphate) (GppNHp) exists in two conformations which are rapidly interconverting with a rate constant of 3200 s-1 at 30 degrees C [Geyer, M., et al. (1996) Biochemistry 35, 10308-10320]. Here we show that Ran complexed with GTP also exists in two conformational states, 1 and 2, which can be directly inferred from the occurrence of two (31)P NMR resonance lines for the gamma-phosphate group of bound GTP. The exchange between the two states is slow on the NMR time scale with a value of <200 s-1 at 5 degrees C for the corresponding first-order rate constants. In wild-type Ran, the equilibrium constant K' between the two states is 0.7 at 278 K, is different for various mutants, and is strongly dependent on the temperature. The standard enthalpy DeltaH degrees and the standard entropy DeltaS degrees for the conformational transitions determined from the NMR spectra are as follows: DeltaH degrees = 37 kJ mol-1 and DeltaS degrees = 130 J mol-1 K-1 for wild-type Ran.GTP. In complex with the Ran-binding protein RanBP1, one of the Ran.GTP conformations (state 2) is stabilized. The interaction of Ran with the guanine nucleotide exchange factor protein RCC1 was also studied by (31)P NMR spectroscopy. In the presence of nucleotide, the ternary complex of Ran.nucleotide.RCC1, an intermediate in the guanine nucleotide exchange reaction, could be observed. A model for the conformational transition of Ran.GTP is proposed where the two states observed are caused by the structural flexibility of the effector loop of Ran; in solution, state 2 resembles the GTP-bound form found in the crystal structure of the Ran-RanBP complex. PMID- 10471275 TI - Protein-protein interactions between the testis brain RNA-binding protein and the transitional endoplasmic reticulum ATPase, a cytoskeletal gamma actin and Trax in male germ cells and the brain. AB - Numerous functions have been proposed for the testis brain RNA-binding protein (TB-RBP) and its human homologue, Translin, ranging from mRNA transport and translational regulation to DNA rearrangement and repair. To gain insight into the likely functions of this 26 kDa protein, immunoprecipitation was used to identify proteins that interact with TB-RBP in mouse cytosolic extracts. Three proteins, the transitional endoplasmic reticulum ATPase, a cytoskeletal gamma actin, and Trax, were specifically immunoprecipitated with an affinity-purified antibody to recombinant mouse TB-RBP. In vitro binding assays with recombinant proteins and EM immunocytochemistry confirm that TB-RBP interacts with the TER ATPase in vitro and in vivo. Confocal microscopy has demonstrated that TB-RBP colocalizes with actin in the cytoplasm of male germ cells. The immunoprecipitation of Trax with TB-RBP confirms a published report demonstrating protein interactions between the two proteins in a yeast two-hybrid assay. These data support the hypothesis that TB-RBP serves as a link in attaching specific mRNAs to cytoskeletal structures and suggests an involvement for the ubiquitously expressed TER ATPase in intracellular and/or intercellular mRNA transport. PMID- 10471276 TI - The EH1 domain of Eps15 is structurally classified as a member of the S100 subclass of EF-hand-containing proteins. AB - The Eps15 homology (EH) domain is a protein-protein interaction module that binds to proteins containing the asparagine-proline-phenylalanine (NPF) or tryptophan/phenylalanine-tryptophan (W/FW) motif. EH domain-containing proteins serve important roles in signaling and processes connected to transport, protein sorting, and organization of subcellular structure. Here, we report the solution structure of the apo form of the EH1 domain of mouse Eps15, as determined by high resolution multidimensional heteronuclear NMR spectroscopy. The polypeptide folds into six alpha-helices and a short antiparallel beta-sheet. Additionally, it contains a long, structured, topologically unique C-terminal loop. Helices 2-5 form two EF-hand motifs. Structural similarity and Ca(2+) binding properties lead to classification of the EH1 domain as a member of the S100 subclass of EF-hand containing proteins, albeit with a unique set of interhelical angles. Binding studies using an eight-residue NPF-containing peptide derived from RAB, the cellular cofactor of the HIV Rev protein, show a hydrophobic peptide-binding pocket formed by conserved tryptophan and leucine residues. PMID- 10471277 TI - Probing the salmeterol binding site on the beta 2-adrenergic receptor using a novel photoaffinity ligand, [(125)I]iodoazidosalmeterol. AB - Salmeterol is a long-acting beta2-adrenergic receptor (beta 2AR) agonist used clinically to treat asthma. In addition to binding at the active agonist site, it has been proposed that salmeterol also binds with very high affinity at a second site, termed the "exosite", and that this exosite contributes to the long duration of action of salmeterol. To determine the position of the phenyl ring of the aralkyloxyalkyl side chain of salmeterol in the beta 2AR binding site, we designed and synthesized the agonist photoaffinity label [(125)I]iodoazidosalmeterol ([125I]IAS). In direct adenylyl cyclase activation, in effects on adenylyl cyclase after pretreatment of intact cells, and in guinea pig tracheal relaxation assays, IAS and the parent drug salmeterol behave essentially the same. Significantly, the photoreactive azide of IAS is positioned on the phenyl ring at the end of the molecule which is thought to be involved in exosite binding. Carrier-free radioiodinated [125I]IAS was used to photolabel epitope-tagged human beta 2AR in membranes prepared from stably transfected HEK 293 cells. Labeling with [(125)I]IAS was blocked by 10 microM (-)-alprenolol and inhibited by addition of GTP gamma S, and [125I]IAS migrated at the same position on an SDS-PAGE gel as the beta 2AR labeled by the antagonist photoaffinity label [125I]iodoazidobenzylpindolol ([125I]IABP). The labeled receptor was purified on a nickel affinity column and cleaved with factor Xa protease at a specific sequence in the large loop between transmembrane segments 5 and 6, yielding two peptides. While the control antagonist photoaffinity label [125I]IABP labeled both the large N-terminal fragment [containing transmembranes (TMs) 1-5] and the smaller C-terminal fragment (containing TMs 6 and 7), essentially all of the [125I]IAS labeling was on the smaller C-terminal peptide containing TMs 6 and 7. This direct biochemical evidence demonstrates that when salmeterol binds to the receptor, its hydrophobic aryloxyalkyl tail is positioned near TM 6 and/or TM 7. A model of IAS binding to the beta 2AR is proposed. PMID- 10471278 TI - Photochemistry of the primary event in short-wavelength visual opsins at low temperature. AB - Two short-wavelength cone opsins, frog (Xenopus laevis) violet and mouse UV, were expressed in mammalian COS1 cells, purified in delipidated form, and studied using cryogenic UV-vis spectrophotometry. At room temperature, the X. laevis violet opsin has an absorption maximum at 426 nm when generated with 11-cis retinal and an absorption maximum of 415 nm when generated with 9-cis-retinal. The frog short-wavelength opsin has two different batho intermediates, one stable at 30 K (lambda(max) approximately 446 nm) and the other at 70 K (lambda(max) approximately 475 nm). Chloride ions do not affect the absorption maximum of the violet opsin. At room temperature, mouse UV opsin has an absorption maximum of 357 nm, while at 70 K, the pigment exhibits a bathochromic shift to 403 nm with distinct vibronic structure and a strong secondary vibronic band at 380 nm. We have observed linear relationships when analyzing the energy difference between the initial and bathochromic intermediates and the normalized difference spectra of the batho-shifted intermediates of rod and cone opsins. We conclude that the binding sites of these pigments change from red to green to violet via systematic shifts in the position of the primary counterion relative to the protonated Schiff base. The mouse UV cone opsin does not fit this trend, and we conclude that wavelength selection in this pigment must operate via a different molecular mechanism. We discuss the possibility that the mouse UV chromophore is initially unprotonated. PMID- 10471279 TI - Saccharomyces cerevisiae Ntg1p and Ntg2p: broad specificity N-glycosylases for the repair of oxidative DNA damage in the nucleus and mitochondria. AB - Saccharomyces cerevisiae possesses two functional homologues (Ntg1p and Ntg2p) of the Escherichia coli endonuclease III protein, a DNA base excision repair N glycosylase with a broad substrate specificity directed primarily against oxidatively damaged pyrimidines. The substrate specificities of Ntg1p and Ntg2p are similar but not identical, and differences in their amino acid sequences as well as inducibility by DNA damaging agents suggest that the two proteins may have different biological roles and subcellular locations. Experiments performed on oligonucleotides containing a variety of oxidative base damages indicated that dihydrothymine, urea, and uracil glycol are substrates for Ntg1p and Ntg2p, although dihydrothymine was a poor substrate for Ntg2p. Vectors encoding Ntg1p green fluorescent protein (GFP) and Ntg2p-GFP fusions under the control of their respective endogenous promoters were utilized to observe the subcellular targeting of Ntg1p and Ntg2p in S. cerevisiae. Fluorescence microscopy of pNTG1 GFP and pNTG2-GFP transformants revealed that Ntg1p localizes primarily to the mitochondria with some nuclear localization, whereas Ntg2p localizes exclusively to the nucleus. In addition, the subcellular location of Ntg1p and Ntg2p confers differential sensitivities to the alkylating agent MMS. These results expand the known substrate specificities of Ntg1p and Ntg2p, indicating that their base damage recognition ranges show distinct differences and that these proteins mediate different roles in the repair of DNA base damage in the nucleus and mitochondria of yeast. PMID- 10471280 TI - Probing the diphosphoglycerate binding pocket of HbA and HbPresbyterian (beta 108Asn --> Lys). AB - HbPresbyterian (beta 108Asn --> Lys, HbP) contains an additional positive charge (per alpha beta dimer) in the middle of the central cavity and exhibits a lower oxygen affinity than wild-type HbA in the presence of chloride. However, very little is known about the molecular origins of its altered functional properties. In this study, we have focused on the beta beta cleft of the Hb tetramer. Recently, we developed an approach for quantifying the ligand binding affinity to the beta-end of the Hb central cavity using fluorescent analogues of the natural allosteric effector 2, 3-diphosphoglycerate (DPG) [Gottfried, D. S., et al. (1997) J. Biol. Chem. 272, 1571-1578]. Time-correlated single-photon counting fluorescence lifetime studies were used to assess the binding of pyrenetetrasulfonate to both HbA and HbP in the deoxy and CO ligation states under acidic and neutral pH conditions. Both the native and mutant proteins bind the probe at a weak binding site and a strong binding site; in all cases, the binding to HbP was stronger than to HbA. The most striking finding was that for HbA the binding affinity varies as follows: deoxy (pH 6.35) > deoxy (pH 7.20) > CO (pH 6.35); however, the binding to HbP is independent of ligation or pH. The mutant oxy protein also hydrolyzes p-nitrophenyl acetate, through a reversible acyl-imidazole pathway linked to the His residues of the beta beta cleft, at a considerably higher rate than does HbA. This implies a perturbation of the microenvironment of these residues at the DPG binding pocket. Structural consequences due to the presence of the new positive charge in the middle of the central cavity have been transmitted to the beta beta cleft of the protein, even in its liganded conformation. This is consistent with a newly described quaternary state (B) for liganded HbPresbyterian and an associated change in the allosteric control mechanism. PMID- 10471281 TI - Retinylidene ligand structure in bovine rhodopsin, metarhodopsin-I, and 10 methylrhodopsin from internuclear distance measurements using 13C-labeling and 1 D rotational resonance MAS NMR. AB - Rhodopsin is the G-protein coupled photoreceptor that initiates the rod phototransduction cascade in the vertebrate retina. Using specific isotope enrichment and magic angle spinning (MAS) NMR, we examine the spatial structure of the C10-C11=C12-C13-C20 motif in the native retinylidene chromophore, its 10 methyl analogue, and the predischarge photoproduct metarhodopsin-I. For the rhodopsin study 11-Z-[10,20-(13)C(2)]- and 11-Z-[11,20-(13)C(2)]-retinal were synthesized and incorporated into bovine opsin while maintaining a natural lipid environment. The ligand is covalently bound to Lys(296) in the photoreceptor. The C10-C20 and C11-C20 distances were measured using a novel 1-D CP/MAS NMR rotational resonance experimental procedure that was specifically developed for the purpose of these measurements [Verdegem, P. J. E., Helmle, M., Lugtenburg, J., and de Groot, H. J. M. (1997) J. Am. Chem. Soc. 119, 169]. We obtain r(10,20) = 0.304 +/- 0.015 nm and r(11,20) = 0.293 +/- 0.015 nm, which confirms that the retinylidene is 11-Z and shows that the C10-C13 unit is conformationally twisted. The corresponding torsional angle is about 44 degrees as indicated by Car Parrinello modeling studies. To increase the nonplanarity in the chromophore, 11 Z-[10,20-(13)C(2)]-10-methylretinal and 11-Z-[(10-CH(3)), 13-(13)C(2)]-10 methylretinal were prepared and incorporated in opsin. For the resulting analogue pigment r(10,20) = 0.347 +/- 0.015 nm and r((10)(-)(CH)()3())(,)(13) = 0.314 +/- 0.015 nm were obtained, consistent with a more distorted chromophore. The analogue data are in agreement with the induced fit principle for the interaction of opsin with modified retinal chromophores. Finally, we determined the intraligand distances r(10,20) and r(11,20) also for the photoproduct metarhodopsin-I, which has a relaxed all-E structure. The results (r(10,20) >/= 0.435 nm and r(11,20) = 0.283 +/- 0.015 nm) fully agree with such a relaxed all-E structure, which further validates the 1-D rotational resonance technique for measuring intraligand distances and probing ligand structure. As far as we are aware, these results represent the first highly precise distance determinations in a ligand at the active site of a membrane protein. Overall, the MAS NMR data indicate a tight binding pocket, well defined to bind specifically only one enantiomer out of four possibilities and providing a steric complement to the chromophore in an ultrafast ( approximately 200 fs) isomerization process. PMID- 10471282 TI - Kinetic description of structural changes linked to membrane import of the colicin E1 channel protein. AB - Upon binding to membranes, the 178-residue colicin E1 C-terminal channel protein forms a steady-state closed-channel intermediate that is a flexible extended two dimensional helical array [Zakharov et al. (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 4282-4287]. Analysis of the kinetics of binding-insertion to liposome membranes of the channel protein, P178, and of changes of spectral parameters associated with structure transitions allowed a correlation of the sequence of tertiary and secondary structure changes with binding-insertion. Binding and insertion were distinguished by use of lipids modified with quenchers of Trp fluorescence attached to lipid headgroups or acyl chains. Secondary and tertiary structure changes were inferred, respectively, from changes in far-UV circular dichroism and relative changes of interresidue distances by fluorescence resonance energy transfer (FRET). "Single Trp" mutants were used in FRET analysis, with the background Tyr contribution determined through use of a "zero Trp" mutant. The sequence of distinguishable events and the pseudo-first-order rate constants under "standard" conditions (large unilamellar vesicles, pH 4.0, I = 0.1 M) was binding (30 +/- 5 s(-)(1)) --> unfolding (12.6 +/- 0.5 s(-)(1)) --> helix elongation (9.0 +/- 1.0 s(-)(1)) --> insertion (6. 6 +/- 0.5 s(-)(1)). Thus, helix elongation on the surface of the membrane can occur after unfolding and does not require insertion. Binding-insertion and structural transitions of P178 occur significantly faster with small unilamellar vesicles. The relevance to general mechanisms of protein import of the structural changes associated with import of the colicin channel is discussed. PMID- 10471283 TI - Ultraviolet laser footprinting of histone H1(0)-four-way junction DNA complexes. AB - We have used a new light footprinting technique to study the interaction of histone H1(0) and a deletion mutant delta CH1(0) (lacking H1(0) COOH-terminal domain) with a synthetic four-way junction DNA. This technique is based on a single 5-ns UV laser pulse and has the ability to map protein-DNA interactions within unperturbed complexes at time scales far faster than molecular rearrangements. We found both H1(0) and delta CH1(0) to affect the photoreactivity of specific guanine residues located on the central part of four way junction DNA. These observations demonstrate specific recognition of H1(0) for the central domain of four-way junction DNA. In addition, histone H1(0) decreases the photoreactivity of selected guanines located some distance from the crossover, indicating specific involvement of the H1(0) COOH-terminal tail with this region. Immunofractionation of delta CH1(0)-four-way DNA junction complexes with monoclonal anti-H1 antibody combined with the UV laser footprinting method demonstrated the existence of two types of delta CH1(0)-four-way DNA junction complexes. PMID- 10471284 TI - Dimer dissociation and thermosensitivity kinetics of the Saccharomyces cerevisiae and human TATA binding proteins. AB - A kinetic analysis of dimer dissociation, TATA DNA binding, and thermal inactivation of the yeast Saccharomyces cerevisiae and human TATA binding proteins (TBP) was conducted. We find that yeast TBP dimers, like human TBP dimers, are slow to dissociate in vitro (t(1/2) approximately 20 min). Mild mutations in the crystallographic dimer interface accelerate the rate of dimer dissociation, whereas severe mutations prevent dimerization. In the presence of excess TATA DNA, which measures the entire active TBP population, dimer dissociation represents the rate-limiting step in DNA binding. These findings provide a biochemical extension to genetic studies demonstrating that TBP dimerization prevents unregulated gene expression in yeast [Jackson-Fisher, A. J., Chitikila, C., Mitra, M., and Pugh, B. F. (1999) Mol. Cell 3, 717-727]. In the presence of vast excesses of TBP over TATA DNA, which measures only a very small fraction of the total TBP, the monomer population in a monomer/dimer equilibrium binds DNA rapidly, which is consistent with a simultaneous binding and bending of the DNA. Under conditions where other studies failed to detect dimers, yeast TBP's DNA binding activity was extremely labile in the absence of TATA DNA, even at temperatures as low as 0 degrees C. Kinetic analyses of TBP instability in the absence of DNA at 30 degrees C revealed that even under fairly stabilizing solution conditions, TBP's DNA binding activity rapidly dissipated with t(1/2) values ranging from 6 to 26 min. TBP's stability appeared to vary with the square root of the TBP concentration, suggesting that TBP dimerization helps prevent TBP inactivation. PMID- 10471285 TI - Sequence and functional-group specificity for cleavage of DNA junctions by RuvC of Escherichia coli. AB - RuvC is the DNA junction-resolving enzyme of Escherichia coli. While the enzyme binds to DNA junctions independently of base sequence, it exhibits considerable sequence selectivity for the phosphodiester cleavage reaction. We have analyzed the sequence specificity using a panel of DNA junctions, measuring the rate of cleavage of each under single-turnover conditions. We have found that the optimal sequence for cleavage can be described by (A approximately T)TT downward arrow(C>G approximately A), where downward arrow denotes the position of backbone scission. Cleavage is fastest when the cleaved phosphodiester linkage is located at the point of strand exchange. However, cleavage is possible one nucleotide 3' of this position when directed by the sequence, with a rate that is 1 order of magnitude slower than the optimal. The maximum sequence discrimination occurs at the central TT in the tetranucleotide site, where any alteration of sequence results in a rate reduction of at least 100-fold and cleavage is undetectable for some changes. However, certain sequences in the outer nucleotides are strongly inhibitory to cleavage. Introduction of base analogues around the cleavage site reveals a number of important functional groups and suggests that major-groove contacts in the center of the tetranucleotide are important for the cleavage process. Since RuvC binds to all the variant junctions with very similar affinity, any contacts affecting the rate of cleavage must be primarily important in the transition state. Introduction of the optimal cleavage sequence into a three-way DNA junction led to relatively efficient cleavage by RuvC, at a rate only 3-fold slower than the optimal four-way junction. This is consistent with a protein-induced alteration in the conformation of the DNA. PMID- 10471286 TI - Effect of nonpolar substitutions of the conserved Phe11 in the fusion peptide of HIV-1 gp41 on its function, structure, and organization in membranes. AB - The fusion domain of the HIV-1 envelope glycoprotein (gp120-gp41) is a conserved hydrophobic region located at the N-terminus of the transmembrane subunit (gp41). A prominent feature of this domain is a conserved five-residue "FLGFL" sequence at positions 8-12. Mutation of the highly conserved Phe(11) to Val (F11V), presumed not to significantly affect the hydrophobicity and the structure of this region, has been shown to decrease the level of syncytium formation and virus infectivity. Here we show that the substitution of Gly for Phe(11) (F11G) reduces cell-cell fusion activity by 80-90%. To determine the effect of these mutations on the properties of the fusion peptide, a 33-residue peptide (WT) identical to the extended fusion domain and its F11V and F11G mutants were synthesized, fluorescently labeled, and studied with respect to their function, structure, and organization in phospholipid membranes. The WT peptide alone induced fusion of both zwitterionic (PC/Chol) and negatively charged (PS/PC/Chol and POPG) vesicles, in contrast to a 23-mer fusion peptide lacking the C-terminal domain which has been shown to be inactive with PC vesicles but able to induce fusion of POPG vesicles which had been preaggragated with Ca(2+) or Mg(2+). The F11V peptide preserved 50% activity, and the F11G peptide was virtually inactive. ATR FTIR spectroscopy indicated similar secondary structure of the peptides in multibilayers that was independent of membrane composition. Furthermore, all the peptides increased the extent of lipid disorder to a similar extent, but the kinetics of amide II H to D exchange was in the following order: F11G > F11V > WT. Fluorescence studies in the presence of membranes, as well as SDS-PAGE, revealed that the WT and F11V peptides self-associate to similar levels while F11G exhibited a decreased level of self-association. The data suggest that the FLGFL motif contributes to the functional organization of the HIV-1 fusion peptide and that the C-terminal domain following the fusion peptide contributes to the membrane fusion process. PMID- 10471287 TI - How fast does the gamma-aminobutyric acid receptor channel open? Kinetic investigations in the microsecond time region using a laser-pulse photolysis technique. AB - The gamma-aminobuytric acid(A) (GABA(A)) receptor is a membrane-bound protein that mediates signal transmission between neurons through formation of chloride ion channels. GABA is the activating ligand, which upon binding to the receptor triggers channel opening in the microsecond time domain and reversible desensitization of the receptor in the millisecond time region. We have investigated the channel-opening mechanism for this receptor in rat hippocampal neurons before the protein desensitizes by using a rapid flow method (cell-flow) with a 10 ms time resolution and a laser-pulse photolysis technique with a approximately 30 micros time resolution to determine the rate and equilibrium constants for channel opening and closing. Two different forms of the receptor, namely, a rapidly and a slowly desensitizing form, exist in the rat hippocampal cells and are characterized by their different rates for desensitization. At 250 microM GABA the rate constant for desensitization was 2.3 +/- 0.4 s(-)(1) for the rapidly desensitizing form and 0.4 +/- 0.1 s(-)(1) for the slowly desensitizing form. The dissociation constant of GABA from the site controlling channel opening was 100 +/- 40 microM for the rapidly desensitizing form and 120 +/- 60 microM for the slowly desensitizing form. The rate constants for channel closing did not differ significantly for the two forms, 85 +/- 20 s(-)(1) for the rapidly desensitizing and 100 +/- 60 s(-)(1) for the slowly desensitizing form. However, the channel-opening rate constant differed by a factor of 3, 1840 +/- 160 s(-)(1) for the rapidly desensitizing and 6700 +/- 330 s(-)(1) for the slowly desensitizing form. This difference in the rate constant for channel opening for the two forms, determined by the laser-pulse photolysis technique, is reflected as a shift in the channel-opening equilibrium constant, which is 7 +/- 5 and 20 +/- 15 for the rapidly and slowly desensitizing forms respectively, determined by the cell-flow method. These constants, together with the concentration of GABA and the concentration of receptor sites in the membrane, determine the number of channels that open as a function of GABA concentration, and the rate at which they open and close. These constants play an important role in determining the rate of the transmembrane ion flux and, therefore, the receptor-controlled changes in transmembrane voltage that trigger signal transmission. PMID- 10471288 TI - Membrane insertion scanning of the human ileal sodium/bile acid co-transporter. AB - Mammalian sodium-dependent bile acid transporters (SBATs) responsible for bile salt uptake across the liver sinusoidal or ileal/renal brush border membrane have been identified and share approximately 35% amino acid sequence identity. Programs for prediction of topology and localization of transmembrane helices identify eight or nine hydrophobic regions for the SBAT sequences as membrane spanning. Analysis of N-linked glycosylation has provided evidence for an exoplasmic N-terminus and a cytoplasmic C-terminus, indicative of an odd number of transmembrane segments. To determine the membrane topography of the human ileal SBAT (HISBAT), an in vitro translation/translocation protocol was employed using three different fusion protein constructs. Individual HISBAT segments were analyzed for signal anchor or stop translocation (stop transfer) activity by insertion between a cytoplasmic anchor (HK M0) or a signal anchor segment (HK M1) and a glycosylation flag (HK beta). To examine consecutive HISBAT sequences, sequential hydrophobic sequences were inserted into the HK M0 vector or fusion vectors were made that included the glycosylated N-terminus of HISBAT, sequential hydrophobic sequences, and the glycosylation flag. Individual signal anchor (SA) and stop transfer (ST) properties were found for seven out of the nine predicted hydrophobic segments (H1, H2, H4, H5, H6, H7, and H9), supporting a seven transmembrane segment model. However, the H3 region was membrane inserted when translated in the context of the native HISBAT flanking sequences. Furthermore, results from translations of sequential constructs ending after H7 provided support for integration of H8. These data provide support for a SBAT transmembrane domain model with nine integrated segments with an exoplasmic N terminus and a cytoplasmic C-terminus consistent with a recent predictive analysis of this transporter topology. PMID- 10471289 TI - Mutant Phe788 --> Leu of the Na+,K+-ATPase is inhibited by micromolar concentrations of potassium and exhibits high Na+-ATPase activity at low sodium concentrations. AB - Mutant Phe788 --> Leu of the rat kidney Na+,K(+)-ATPase was expressed in COS cells to active-site concentrations between 40 and 60 pmol/mg of membrane protein. Analysis of the functional properties showed that the discrimination between Na+ and K+ on the two sides of the system is severely impaired in the mutant. Micromolar concentrations of K+ inhibited ATP hydrolysis (K(0.5) for inhibition 107 microM for the mutant versus 76 mM for the wild-type at 20 mM Na+), and at 20 mM K+, the molecular turnover number for Na+,K(+)-ATPase activity was reduced to 11% that of the wild-type. This inhibition was counteracted by Na+ in high concentrations, and in the total absence of K+, the mutant catalyzed Na(+)-activated ATP hydrolysis ("Na(+)-ATPase activity") at an extraordinary high rate corresponding to 86% of the maximal Na+,K(+)-ATPase activity. The high Na(+) ATPase activity was accounted for by an increased rate of K(+)-independent dephosphorylation. Already at 2 mM Na+, the dephosphorylation rate of the mutant was 8-fold higher than that of the wild-type, and the maximal rate of Na(+) induced dephosphorylation amounted to 61% of the rate of K(+)-induced dephosphorylation. The cause of the inhibitory effect of K+ on ATP hydrolysis in the mutant was an unusual stability of the K(+)-occluded E2(K2) form. Hence, when E2(K2) was formed by K+ binding to unphosphorylated enzyme, the K(0.5) for K+ occlusion was close to 1 microM in the mutant versus 100 microM in the wild-type. In the presence of 100 mM Na+ to compete with K+ binding, the K(0.5) for K+ occlusion was still 100-fold lower in the mutant than in the wild-type. Moreover, relative to the wild-type, the mutant exhibited a 6-7-fold reduced rate of release of occluded K+, a 3-4-fold increased apparent K+ affinity in activation of the pNPPase reaction, a 10-11-fold lower apparent ATP affinity in the Na+,K(+) ATPase assay with 250 microM K+ present (increased K(+)-ATP antagonism), and an 8 fold reduced apparent ouabain affinity (increased K(+)-ouabain antagonism). PMID- 10471290 TI - Targeted cross-linking of a molten globule form of acetylcholinesterase by the virucidal agent hypericin. AB - The natural product hypericin is a photosensitive polycyclic aromatic dione compound, which has been widely investigated because of its virucidal and antitumor properties. Although it has been suggested that singlet oxygen or a radical species might be responsible for its biological action, its mechanism of action remains unknown. Due to its amphiphilic characteristics, we considered the possibility that it might interact preferentially with partially unfolded proteins which exhibit exposed hydrophobic surfaces. We here demonstrate that hypericin binds to a molten globule species generated from Torpedo acetylcholinesterase, but not to the corresponding native enzyme. Irradiation with visible light, under aerobic conditions, causes chemical cross-linking of the catalytic subunits, to dimers and heavier species, under conditions where no cross-linking is observed for the native enzyme. Both anaerobiosis and sodium azide greatly reduce the extent of cross-linking, suggesting that singlet oxygen is responsible for the phenomenon. This agrees with our observation, using spin traps, that mainly singlet oxygen is produced by the complex of hypericin with the molten globule of acetylcholinesterase. Cross-linking is enhanced in the presence of liposomes to which the molten globule of acetylcholinesterase is quantitatively adsorbed. This may be due to high local concentrations of both hypericin and the protein resulting in close proximity, and hence in a high yield of cross-linking. Molten globule species are believed to be intermediates in both protein folding and translocation through biological membranes. Thus, hypericin may serve as a valuable tool for trapping such intermediates. This might also explain its therapeutic effectiveness toward virus-infected or tumor cells. PMID- 10471291 TI - Inhibition of nicotinic acetylcholine receptor by philanthotoxin-343: kinetic investigations in the microsecond time region using a laser-pulse photolysis technique. AB - The mechanism of inhibition of a nicotinic acetylcholine receptor (nAChR) in BC(3)H1 muscle cells by philanthotoxin-343 (PhTX-343), a synthetic analogue of philanthotoxin-433, a wasp toxin, was investigated using a laser-pulse photolysis technique with microsecond time resolution and in a carbamoylcholine concentration range of 20-100 microM and PhTX-343 concentration range of 0-200 microM. The rate constant for nAChR channel opening determined by the chemical kinetic techniques decreased with increasing PhTX-343 concentration, whereas there was no significant effect on the rate constant for channel closing. The resulting decrease in the channel-opening equilibrium constant accounted quantitatively for the inhibition of the receptor by the toxin. Single-channel current measurements suggest an additional step in which the open channel:inhibitor complex isomerizes to a nonconducting receptor form. Cell-flow experiments with a time resolution of 10 ms indicate that this isomerization step is only important at very high inhibitor concentrations. The inhibitor binds to the open-channel receptor form, with an affinity that is at least 5 times smaller than that for the closed-channel form. This indicates that receptor inhibition mainly involves the binding of PhTX-343 to the closed-channel form of the receptor. PhTX-343, and an analogue of this polyamine, had no effect when applied to the inside of the cell membrane. However, there was significant inhibition of the nAChR when these compounds were applied to the outside of the cell membrane, indicating an extracellular site for inhibition. Furthermore, increasing the transmembrane potential results in a decrease in the ability of PhTX-343 to inhibit the receptor. This observation is related to the voltage dependence of the effect of PhTX-343 on the rate constant for nAChR channel opening with increasing transmembrane voltage (-60 to 50 mV). This suggests that the voltage dependence of inhibition mainly reflects the effect of transmembrane voltage on the rate constant of channel opening and, therefore, the channel-opening equilibrium constant. PhTX-343 competes with cocaine and procaine for its binding site. The finding that this toxin, which binds to a common inhibitory site with compounds such as cocaine, exerts its effect by decreasing the channel-opening equilibrium constant suggests an approach for the development of therapeutic agents. A compound that binds to this regulatory site but does not affect the channel-opening equilibrium constant may be developed. Such a compound can displace an abused drug such as cocaine and thereby alleviate the toxic effect of this compound on the organism. PMID- 10471292 TI - A complex family of highly heterogeneous and internally repetitive hyperactive antifreeze proteins from the beetle Tenebrio molitor. AB - We have previously identified a Thr- and Cys-rich thermal hysteresis (antifreeze) protein (THP) in the beetle Tenebrio molitor that has 10-100 times the freezing point depression activity of fish antifreeze proteins. Because this 8.4 kDa protein is significantly different in its properties from THP preparations previously reported from this insect, a thorough search was undertaken for other antifreeze types. Many active proteins were observed, but all appeared to be isoforms of the THP that differed in their number of 12-amino acid repeats (consensus sequence CTxSxxCxxAxT), amino acid substitutions, and N-linked glycosylation. Mass spectral analysis has matched most of these isoforms with cDNA sequences of 17 different clones from a larval fat body library that encode eight different mature THPs containing 84, 96, or 120 amino acids. Genomic Southern blots suggest there may be 30-50 tightly linked copies of the gene, which is a signature consistently seen with unrelated fish antifreeze protein genes, and one that has been associated with the need to rapidly increase gene product in response to climate change. A three-dimensional model is proposed for the fully disulfide-bonded structure of T. molitor THP, which can accommodate addition or deletion of 12-amino acid repeats. The structure is a beta-helix that places most of the Thr in a regular array on one side of the protein to form a putative ice-binding surface. PMID- 10471293 TI - Redesign of cytochrome c peroxidase into a manganese peroxidase: role of tryptophans in peroxidase activity. AB - Trp191Phe and Trp51Phe mutations have been introduced into an engineered cytochrome c peroxidase (CcP) containing a Mn(II)-binding site reported previously (MnCcP; see Yeung, B. K.-S., et al. (1997) Chem. Biol. 5, 215-221). The goal of the present study is to elucidate the role of tryptophans in peroxidase activity since CcP contains both Trp51 and Trp191 while manganese peroxidase (MnP) contains phenylalanine residues at the corresponding positions. The presence of Trp191 in CcP allows formation of a unique high-valent intermediate containing a ferryl oxo and tryptophan radical called compound I'. The absence of a tryptophan residue at this position in MnP is the main reason for the formation of an intermediate called compound I which contains a ferryl oxo and porphyrin pi-cation radical. In this study, we showed that introduction of the Trp191Phe mutation to MnCcP did not improve MnP activity (specific activity: MnCcP, 0.750 micromol min-1 mg-1; MnCcP(W191F), 0.560 micromol min-1 mg 1. k(cat)/K(m): MnCcP, 0.0517 s-1 mM-1; MnCcP(W191F), 0.0568 s-1 mM-1) despite the fact that introduction of the same mutation to WTCcP caused the formation of a transient compound I (decay rate, 60 s-1). However, introducing both the Trp191Phe and Trp51Phe mutations not only resulted in a longer lived compound I in WTCcP (decay rate, 18 s-1), but also significantly improved MnP activity in MnCcP (MnCcP(W51F, W191F): specific activity, 8.0 micromol min-1 mg-1; k(cat)/K(m), 0. 599 s-1 mM-1). The increase in activity can be attributed to the Trp51Phe mutation since MnCcP(W51F) showed significantly increased MnP activity relative to MnCcP (specific activity, 3.2 micromol min-1 mg-1; k(cat)/K(m), 0.325 s-1 mM-1). As with MnP, the activity of MnCcP(W51F, W191F) was found to increase with decreasing pH. Our results demonstrate that, while the Trp191Phe and Trp51Phe mutations both play important roles in stabilizing compound I, only the Trp51Phe mutation contributes significantly to increasing the MnP activity because this mutation increases the reactivity of compound II, whose oxidation of Mn(II) is the rate-determining step in the reaction mechanism. PMID- 10471294 TI - Inhibition of the aminopeptidase from Aeromonas proteolytica by aliphatic alcohols. Characterization of the hydrophobic substrate recognition site. AB - Seven aliphatic and two aromatic alcohols were tested as reporters of the substrate selectivity of the aminopeptidase from Aeromonas proteolytica (AAP). This series of alcohols was chosen to systematically probe the effect of carbon chain length, steric bulk, and inhibitor shape on the inhibition of AAP. Initially, however, the question of whether AAP is denatured in the presence of aliphatic alcohols was addressed. On the basis of circular dichroism (CD), electronic absorption, and fluorescence spectra, the secondary structure of AAP, with and without added aliphatic alcohols, was unchanged. These data clearly indicate that AAP is not denatured in aliphatic alcohols, even up to concentrations of 20% (v/v). All of the alcohols studied were competitive inhibitors of AAP with K(i) values between 860 and 0.98 mM. The clear trend in the data was that as the carbon chain length increases from one to four, the K(i) values increase. Branching of the carbon chains also increases the K(i) values, but large bulky groups, such as that found in tert-butyl alcohol, do not inhibit AAP as well as leucine analogues, such as 3-methyl-1-butanol. The competitive nature of the inhibition indicates that the substrate and each alcohol studied are mutually exclusive due to binding at the same site on the enzyme. On the basis of EPR and electronic absorption data for Co(II)-substituted AAP, none of the alcohols studied binds to the dinuclear metallo-active site of AAP. Thus, reaction of the inhibitory alcohols with the catalytic metal ions cannot constitute the mechanism of inhibition. Combination of these data suggests that each of these inhibitors bind only to the hydrophobic pocket of AAP and, consequently, block the binding of substrate. Thus, the first step in peptide hydrolysis is the recognition of the N-terminal amino acid side chain by the hydrophobic pocket adjacent to the dinuclear active site of AAP. PMID- 10471295 TI - Change of nucleotide specificity and enhancement of catalytic efficiency in single point mutants of Vibrio harveyi aldehyde dehydrogenase. AB - The fatty aldehyde dehydrogenase from the luminescent bacterium, Vibrio harveyi (Vh-ALDH), is unique with respect to its high specificity for NADP(+) over NAD(+). By mutation of a single threonine residue (Thr175) immediately downstream of the beta(B) strand in the Rossmann fold, the nucleotide specificity of Vh-ALDH has been changed from NADP(+) to NAD(+). Replacement of Thr175 by a negatively charged residue (Asp or Glu) resulted in an increase in k(cat)/K(m) for NAD(+) relative to that for NADP(+) of up to 5000-fold due to a decrease for NAD(+) and an increase for NADP(+) in their respective Michaelis constants (K(a)). Differential protection by NAD(+) and NADP(+) against thermal inactivation and comparison of the dissociation constants of NMN, 2'-AMP, 2'5'-ADP, and 5'-AMP for these mutants and the wild-type enzyme clearly support the change in nucleotide specificity. Moreover, replacement of Thr175 with polar residues (N, S, or Q) demonstrated that a more efficient NAD(+)-dependent enzyme T175Q could be created without loss of NADP(+)-dependent activity. Analysis of the three-dimensional structure of Vh-ALDH with bound NADP(+) showed that the hydroxyl group of Thr175 forms a hydrogen bond to the 2'-phosphate of NADP(+). Replacement with glutamic acid or glutamine strengthened interactions with NAD(+) and indicated why threonine would be the preferred polar residue at the nucleotide recognition site in NADP(+)-specific aldehyde dehydrogenases. These results have shown that the size and the structure of the residue at the nucleotide recognition site play the key roles in differentiating between NAD(+) and NADP(+) interactions while the presence of a negative charge is responsible for the decrease in interactions with NADP(+) in Vh-ALDH. PMID- 10471296 TI - Partial glycosylation at asparagine-2181 of the second C-type domain of human factor V modulates assembly of the prothrombinase complex. AB - Thrombin-activated factor Va exists as two isoforms, factor Va(1) and factor Va(2), which differ in the size of their light chains and their affinity for biological membranes. The heterogeneity of the light chain remained following incubation of factor Va with N-glycanase. However, we found that the factor V C2 domain, which contains a single potential glycosylation site at Asn-2181, was partially glycosylated when expressed in COS cells. To confirm the structural basis for factor Va(1) and factor Va(2), we mutated Asn-2181 to glutamine (N2181Q) and expressed this mutant using a B domain deletion construct (rHFV des B) in COS cells. Thrombin activation of N2181Q released a light chain with mobility identical to that of factor Va(2) on SDS-PAGE. The functional properties of purified N2181Q were similar to those of factor Va(2) in prothrombinase assays carried out in the presence of limiting concentrations of phosphatidylserine. The binding of human factor Va(1) and factor Va(2) to 75:25 POPC/POPS vesicles was also investigated in equilibrium binding assays using proteins containing a fluorescein-labeled heavy chain. The affinity of human factor Va(2) binding to POPC/POPS vesicles was approximately 3-fold higher than that of factor Va(1). These results indicate that partial glycosylation of factor V at asparagine-2181 is the structural basis of the light chain doublet and that the presence of this oligosaccharide reduces the affinity of factor Va for biological membranes. PMID- 10471297 TI - Elucidation of a novel polypeptide cross-link involving 3-hydroxykynurenine. AB - 3-Hydroxykynurenine, a metabolite of tryptophan, is a powerful antioxidant and neurotoxin. The neurotoxicity results from the oxidation of 3-hydroxykynurenine, and hydroxyl radicals, formed via H(2)O(2), may also be implicated [Okuda, S., Nishiyama, N., Saito, H. , and Katsuki, H. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 12553-12558]. Oxidation of o-aminophenols, such as 3-hydroxykynurenine, also results in the formation of highly reactive quinonimines. Thus, one possible consequence of 3-hydroxykynurenine oxidation may be covalent modification of cellular macromolecules. Such a process could contribute to the neurotoxicity and may potentially be important in other tissues, such as the human lens, where 3 hydroxykynurenine functions as a UV filter. In this work, we demonstrate that 3 hydroxykynurenine can bind to protein amino groups and, further, that under oxidative conditions, 3-hydroxykynurenine can function to cross-link polypeptide chains. The structure of the cross-linked moiety, using the peptide glycyllysine, has been elucidated. The cross-link, which is both colored and fluorescent, involves the peptide alpha-amino groups. Proteins modified by 3-hydroxykynurenine become colored and fluorescent as well as cross-linked. LC-MS studies indicate that the cross-link is also present in gamma-crystallin, following incubation of this lens protein in the presence of 3-hydroxykynurenine. Similar posttranslational modifications of lens proteins accompany cataract formation, and knowledge of the precise mode of reaction of 3-hydroxykynurenine with proteins will assist in determining if 3-hydroxykynurenine is involved in degenerative conditions in which oxidation of such aminophenols is implicated. PMID- 10471298 TI - Observation of the protonated semiquinone intermediate in isolated reaction centers from Rhodobacter sphaeroides: implications for the mechanism of electron and proton transfer in proteins. AB - A proton-activated electron transfer (PAET) mechanism, involving a protonated semiquinone intermediate state, had been proposed for the electron-transfer reaction k(2)AB [Q(A)(-)(*)Q(B)(-)(*) + H(+) <--> Q(A)(-)(*)(Q(B)H)(*) --> Q(A)(Q(B)H)(-)] in reaction centers (RCs) from Rhodobacter sphaeroides [Graige, M. S., Paddock, M. L., Bruce, M. L., Feher, G., and Okamura, M. Y. (1996) J. Am. Chem. Soc. 118, 9005-9016]. Confirmation of this mechanism by observing the protonated semiquinone (Q(B)H)(*) had not been possible, presumably because of its low pK(a). By replacing the native Q(10) in the Q(B) site with rhodoquinone (RQ), which has a higher pK(a), we were able to observe the (Q(B)H)(*) state. The pH dependence of the semiquinone optical spectrum gave a pK(a) = 7.3 +/- 0.2. At pH < pK(a), the observed rate for the reaction was constant and attributed to the intrinsic electron-transfer rate from Q(A)(-)(*) to the protonated semiquinone (i.e., k(2)AB = k(ET)(RQ) = 2 x 10(4) s(-)(1)). The rate decreased at pH > pK(a) as predicted by the PAET mechanism in which fast reversible proton transfer precedes rate-limiting electron transfer. Consequently, near pH 7, the proton transfer rate k(H) >> 10(4) s(-)(1). Applying the two step mechanism to RCs containing native Q(10) and taking into account the change in redox potential, we find reasonable values for the fraction of (Q(B)H)(*) congruent with 0.1% (consistent with a pK(a)(Q(10)) of approximately 4.5) and k(ET)(Q(10)) congruent with 10(6) s(-)(1). These results confirm the PAET mechanism in RCs with RQ and give strong support that this mechanism is active in RCs with Q(10) as well. PMID- 10471299 TI - Understanding protein lids: kinetic analysis of active hinge mutants in triosephosphate isomerase. AB - In previous work we tested what three amino acid sequences could serve as a protein hinge in triosephosphate isomerase [Sun, J., and Sampson, N. S. (1998) Protein Sci. 7, 1495-1505]. We generated a genetic library encoding all 8000 possible 3 amino acid combinations at the C-terminal hinge and selected for those combinations of amino acids that formed active mutants. These mutants were classified into six phylogenetic families. Two families resembled wild-type hinges, and four families represented new types of hinges. In this work, the kinetic characteristics and thermal stabilities of mutants representing each of these families were determined in order to understand what properties make an efficient protein hinge, and why all of the families are not observed in nature. From a steady-state kinetic analysis of our mutants, it is clear that the partitioning between protonation of intermediate to form product and intermediate release from the enzyme surface to form methylglyoxal (a decomposition product) is not affected. The two most impaired mutants undergo a change in rate-limiting step from enediol formation to dihydroxyacetone phosphate binding. Thus, it appears that k(cat)/K(m)'s are reduced relative to wild type as a result of slower Michaelis complex formation and dissociation, rather than increased loop opening speed. PMID- 10471300 TI - Arginine 177 is involved in Mn(II) binding by manganese peroxidase. AB - Site-directed mutations R177A and R177K in the gene encoding manganese peroxidase isozyme 1 (mnp1) from Phanerochaete chrysosporium were generated. The mutant enzymes were expressed in P. chrysosporium during primary metabolic growth under the control of the glyceraldehyde-3-phosphate dehydrogenase gene promoter, purified to homogeneity, and characterized by spectroscopic and kinetic methods. The UV-vis spectra of the ferric and oxidized states and resonance Raman spectra of the ferric state were similar to those of the wild-type enzyme, indicating that the heme environment was not significantly affected by the mutations at Arg177. Apparent K(m) values for Mn(II) were approximately 20-fold greater for the R177A and R177K MnPs than for wild-type MnP. However, the apparent K(m) values for the substrates, H(2)O(2) and ferrocyanide, and the k(cat) values for Mn(II) and ferrocyanide oxidation were similar to those of the wild-type enzyme. The second-order rate constants for compound I (MnPI) reduction of the mutant MnPs by Mn(II) were approximately 10-fold lower than for wild-type MnP. In addition, the K(D) values calculated from the first-order plots of MnP compound II (MnPII) reduction by Mn(II) for the mutant enzymes were approximately 22-fold greater than for wild-type MnP. In contrast, the first-order rate constants for MnPII reduction by Mn(II) were similar for the mutant and wild-type MnPs. Furthermore, second-order rate constants for the wild-type and mutant enzymes for MnPI formation, for MnPI reduction by bromide, and for MnPI and MnPII reduction by ferrocyanide were not significantly changed. These results indicate that both the R177A and R177K mutations specifically affect the binding of Mn, whereas the rate of electron transfer from Mn(II) to the oxidized heme apparently is not affected. PMID- 10471301 TI - Regulation of constitutive gene expression through interactions of Sp1 protein with the nuclear aryl hydrocarbon receptor complex. AB - The region of residues -145 to -119 (CD/L) of the cathepsin D gene promoter contains a GC-rich motif that binds Sp1 protein and an adjacent pentanucleotide (CACGC) that corresponds to the core sequence of a dioxin responsive element (DRE) and binds the aryl hydrocarbon receptor (AhR)-AhR nuclear translocator (Arnt) complex. This Sp1(N)(4)DRE(core) motif has been identified in promoters of several genes in which Sp1 plays an important role in basal gene expression. In transient transfection assays with MCF-7 human breast cancer cells using wild type pCD/L and constructs mutated in the core DRE (pCD/L(m1)) and Sp1 (pCD/L(m2)) sites, it was shown that both motifs were required for maximal basal activity. The requirements for AhR-Arnt interactions with Sp1 protein for maximal activity of pCD/L were confirmed in wild-type MCF-7 and Hepa 1c1c7 cells and Arnt deficient Hepa 1c1c7 cells using antisense Arnt and Arnt expression plasmids. The functional interactions of Sp1 with AhR-Arnt were paralleled by physical interactions showing that AhR-Arnt and Sp1 proteins were co-immunoprecipitated and AhR-Arnt enhanced Sp1-[(32)P]CD/L binding in electrophoretic mobility shift assays. The physical and functional interactions of Sp1 with AhR-Arnt proteins bound to the Sp1(N)(4)DRE(core) motif were also dependent on the proximity of these sites, and both the activity and the extent of Sp1-DNA binding decreased as the number of intervening nucleotides increased from 4 to 20. These studies show that regulation of basal expression of some genes by Sp1 may also require interactions with AhR-Arnt. PMID- 10471302 TI - Isothiazolone derivatives selectively inhibit telomerase from human and rat cancer cells in vitro. AB - The telomere hypothesis postulates stabilization of telomere length and telomerase activation as key events in cellular immortalization and carcinogeneses. Accordingly, telomerase has been suggested as a novel and highly selective target for design of antitumor drugs. Screening of a chemical library including 16 000 synthetic compounds yielded six that strongly inhibited telomerase activity in extracts of cultured human cells, including four isothiazolone derivatives and two unrelated compounds. The most potent inhibitor was 2-[3-(trifluoromethyl)phenyl]isothiazolin-3-one (TMPI), a concentration of 1.0 microM inhibited telomerase activity by 50% according to a telomere repeat amplification protocol (TRAP) assay. Analysis using partially purified telomerase from AH7974 rat hepatoma cells demonstrated noncompetitive inhibition with the telomere-repeat primer and mixed inhibition with the dNTPs; the inhibition constant was 2.5 microM. TMPI did not inhibit eukaryotic DNA polymerase alpha, beta, or human immunodeficiency virus reverse transcriptase (HIV RT). Thus, inhibition by TMPI was highly selective for telomerase. Inhibition by TMPI was quenched by 1 mM of dithiothreitol or glutathione, suggesting that TMPI inhibits telomerase by acting at a cysteine residue. TMPI inhibition of this enzyme may find application as an antineoplastic agent. PMID- 10471303 TI - Bizelesin, a bifunctional cyclopropylpyrroloindole alkylating agent, inhibits simian virus 40 replication in trans by induction of an inhibitor. AB - Bizelesin, a bifunctional DNA minor groove alkylating agent, inhibits both cellular and viral (SV40) DNA replication in whole cells. Bizelesin inhibition of SV40 DNA replication was analyzed in SV40-infected cells, using two-dimensional (2D) neutral agarose gel electrophoresis, and in a cell-free SV40 DNA replication assay. Within 1 h of bizelesin addition to infected cells, a similar rapid decrease in both the level of SV40 replication intermediates and replication activity was observed, indicating inhibition of initiation of SV40 DNA replication. However, prolonged bizelesin treatment (>/=2 h) was associated with a reduced extent of elongation of SV40 replicons, as well as the appearance on 2D gels of intense spots, suggestive of replication pause sites. Inhibition of elongation and induction of replication pause sites may result from the formation of bizelesin covalent bonds on replicating SV40 molecules. The level of in vitro replication of SV40 DNA also was reduced when extracts from bizelesin-treated HeLa cells were used. This effect was not dependent upon the formation of bizelesin covalent bonds with the template DNA. Mixing experiments, using extracts from control and bizelesin-treated cells, indicated that reduced DNA replication competence was due to the presence of a trans-acting DNA replication inhibitor, rather than to decreased levels or inactivation of essential replication factor(s). PMID- 10471304 TI - M-side electron transfer in reaction center mutants with a lysine near the nonphotoactive bacteriochlorophyll. AB - We report the primary charge separation events in a series of Rhodobacter capsulatus reaction centers (RCs) that have been genetically modified to contain a lysine near the bacteriochlorophyll molecule, BChl(M), on the nonphotoactive M side of the RC. Using wild type and previously constructed mutants as templates, we substituted Lys for the native Ser residue at position 178 on the L polypeptide to make the S(L178)K single mutant, the S(L178)K/G(M201)D and S(L178)K/L(M212)H double mutants, and the S(L178)K/G(M201)D/L(M212)H triple mutant. In the triple mutant, the decay of the photoexcited primary electron donor (P) occurs with a time constant of 15 ps and is accompanied by 15% return to the ground state, 62% electron transfer to the L-side bacteriopheophytin, BPh(L), and 23% electron transfer to the M-side analogue, BPh(M). The data supporting electron transfer to the M-side include bleaching of the Q(X) band of BPh(M) at 528 nm and a spectrally and kinetically resolved anion band with a maximum at 640 nm assigned to BPh(M)(-). The decay of these features and concomitant approximately 20% decay of bleaching of the 850 nm band of P give a P(+)BPh(M)(-) lifetime on the order of 1-2 ns that reflects deactivation to give the ground state. These data and additional findings are compared to those from parallel experiments on the G(M201)D/L(M212)H double mutant, in which 15% electron transfer to BPh(M) has been reported previously and is reproduced here. We also compare the above results with the primary electron-transfer processes in S(L178)K, S(L178)K/G(M201)D, and S(L178)K /L(M212)H RCs and with those for the L(M212)H and G(M201)D single mutants and wild-type RCs. The comparison of extensive results that track the primary events in these eight RCs helps to elucidate key factors underlying the directionality and high yield of charge separation in the bacterial photosynthetic RC. PMID- 10471305 TI - Plastocyanin binding to photosystem I as a function of the charge state of the metal ion: effect of metal site conformation. AB - The binding of Ag- and Cd-substituted plastocyanin to reduced photosystem 1 of spinach has been studied through the rotational correlation time of plastocyanin measured by the technique of perturbed angular correlation of gamma-rays (PAC). Ag and Cd are used as models for native Cu(I) and Cu(II), respectively. A dissociation constant of 5 microM was found for Ag-plastocyanin, whereas the dissociation constant was at least 24 times higher for Cd-plastocyanin. PAC was further used to characterize the structure of the metal site of Cd- and Ag plastocyanin. The Cd spectra are characteristic of a planar configuration of one cysteine and two histidines. However, the spectra show an unusual peak broadening and a high degree of internal motion, interpreted as motion of one of the histidines within the plane. (111)Ag decays to (111)Cd, followed by the emission of two gamma-rays used for the PAC experiment. The (111)Ag PAC spectra indicate that one of the coordinating histidines has a different position in the Ag protein than in the Cd protein but that the decay of Ag to Cd causes a relaxation of the position of this histidine to the position in the Cd protein within 20 ns. Binding of Ag-plastocyanin to photosystem I stabilized the Ag metal site structure so that no relaxation was observed on a time scale of 100 ns. This stabilization of the Ag structure upon binding indicates that the metal site structure is involved in regulating how the dissociation constant for plastocyanin depends on the charge of the metal ion. PMID- 10471306 TI - Conformational heterogeneity of the bacteriopheophytin electron acceptor HA in reaction centers from Rhodopseudomonas viridis revealed by Fourier transform infrared spectroscopy and site-directed mutagenesis. AB - The light-induced Fourier transform infrared (FTIR) difference spectra corresponding to the photoreduction of either the HA bacteriopheophytin electron acceptor (HA-/HA spectrum) or the QA primary quinone (QA-/QA spectrum) in photosynthetic reaction centers (RCs) of Rhodopseudomonas viridis are reported. These spectra have been compared for wild-type (WT) RCs and for two site-directed mutants in which the proposed interactions between the carbonyls on ring V of HA and the RC protein have been altered. In the mutant EQ(L104), the putative hydrogen bond between the protein and the 9-keto C=O of HA should be affected by changing Glu L104 to a Gln. In the mutant WF(M250), the van der Waals interactions between Trp M250 and the 10a-ester C=O of HA should be modified. The characteristic effects of both mutations on the FTIR spectra support the proposed interactions and allow the IR modes of the 9-keto and 10a-ester C=O of HA and HA- to be assigned. Comparison of the HA-/HA and QA-/QA spectra leads us to conclude that the QA-/QA IR signals in the spectral range above 1700 cm-1 are largely dominated by contributions from the electrostatic response of the 10a-ester C=O mode of HA upon QA photoreduction. A heterogeneity in the conformation of the 10a ester C=O mode of HA in WT RCs, leading to three distinct populations of HA, appears to be related to differences in the hydrogen-bonding interactions between the carbonyls of ring V of HA and the RC protein. The possibility that this structural heterogeneity is related to the observed multiexponential kinetics of electron transfer and the implications for primary processes are discussed. The effect of 1H/2H exchange on the QA-/QA spectra of the WT and mutant RCs shows that neither Glu L104 nor any other exchangeable carboxylic residue changes appreciably its protonation state upon QA reduction. PMID- 10471307 TI - Solution structure of contryphan-R, a naturally occurring disulfide-bridged octapeptide containing D-tryptophan: comparison with protein loops. AB - Contryphan-R is a disulfide-constrained octapeptide containing a D-tryptophan that was isolated recently from venom of the cone shell Conus radiatus. The polypeptide is present in two forms in solution due to cis-trans isomerization at hydroxyproline 3. The solution structure of the major form of this unusual polypeptide, determined from NMR data, consists of a well-defined fold containing a non-hydrogen-bonded chain reversal from Gly1 to Glu5, which includes a cis hydroxyproline and a D-Trp, and a type I beta-turn from Glu5 to Cys8. The presence of a putative salt bridge between the Glu5 carboxyl group and the N terminal ammonium group is investigated by using various solvation models during energy minimization and is compared with the results of a pH titration. A comparison of the structure of contryphan-R with other cyclic peptide structures highlights some of the key structural determinants of these peptides and suggests that the contryphan-R fold could be exploited as a scaffold onto which unrelated protein binding surfaces could be grafted. Comparison with small disulfide bridged loops in larger proteins shows that contryphan-R is similar to a commonly occurring loop structure found in proteins. PMID- 10471308 TI - Raman spectroscopic study on the copper(II) binding mode of prion octapeptide and its pH dependence. AB - The cellular form of prion protein is a precursor of the infectious isoform, which causes fatal neurodegenerative diseases through intermolecular association. One of the characteristics of the prion protein is a high affinity for Cu(II) ions. The site of Cu(II) binding is considered to be the N-terminal region, where the octapeptide sequence PHGGGWGQ repeats 4 times in tandem. We have examined the Cu(II) binding mode of the octapeptide motif and its pH dependence by Raman and absorption spectroscopy. At neutral and basic pH, the single octapeptide PHGGGWGQ forms a 1:1 complex with Cu(II) by coordinating via the imidazole N pi atom of histidine together with two deprotonated main-chain amide nitrogens in the triglycine segment. A similar 1:1 complex is formed by each octapeptide unit in (PHGGGWGQ)2 and (PHGGGWGQ)4. Under weakly acidic conditions (pH approximately 6), however, the Cu(II)-amide- linkages are broken and the metal binding site of histidine switches from N pi to N tau to share a Cu(II) ion between two histidine residues of different peptide chains. The drastic change of the Cu(II) binding mode on going from neutral to weakly acidic conditions suggests that the micro environmental pH in the brain cell regulates the Cu(II) affinity of the prion protein, which is supposed to undergo pH changes in the pathway from the cell surface to endosomes. The intermolecular His(N tau)-Cu(II)-His(N tau) bridge may be related to the aggregation of prion protein in the pathogenic form. PMID- 10471309 TI - Plasmin cleavage of the amyloid beta-protein: alteration of secondary structure and stimulation of tissue plasminogen activator activity. AB - Cerebrovascular amyloid beta-protein (A beta) deposition, a key pathological feature of Alzheimer's disease and hereditary cerebral hemorrhage with amyloidosis Dutch-type, can lead to intracerebral hemorrhage; however, the mechanism for this remains unclear. Assembled A beta is a potent stimulator of tissue-type plasminogen activator (tPA) in vitro. Herein, we investigated the stimulation of tPA by freshly solubilized A beta 1-40. The rate of tPA stimulation by A beta 1-40 increased dramatically over time, suggesting that A beta may be altered during the course of the reaction. SDS-PAGE analysis showed that A beta 1-40 was cleaved during the course of the reaction. Subsequent studies showed that it was plasmin, the product of tPA activation of plasminogen, that specifically cleaved A beta 1-40 in the amino terminal region between Arg5 and His6. Plasmin effectively cleaved a chromogenic substrate corresponding to this cleavage site in A beta. Circular dichroism spectral analysis showed that A beta 6-40 adopted a strong beta-sheet secondary structure. This truncated A beta 6-40 peptide was a potent stimulator of tPA in vitro. Our results indicate that beta-sheet secondary structure of A beta, which can be promoted by plasmin cleavage, stimulates tPA activity. These findings suggest that pathologic interactions between A beta, tPA, and plasmin in the cerebral vessel wall could result in excessive proteolysis contributing to intracerebral hemorrhages. PMID- 10471310 TI - Helix 8 and helix 10 are involved in substrate recognition in the rat monocarboxylate transporter MCT1. AB - Transport of lactate, pyruvate, and the ketone bodies, acetoacetate and beta hydroxybutyrate, is mediated in many mammalian cells by the monocarboxylate transporter MCT1. To be accepted as a substrate, a carboxyl group and an unpolar side chain are necessary. Site-directed mutagenesis of the rat MCT1 was used to identify residues which are involved in substrate recognition. Helices 8 and 10 but not helix 9 were found to contain critical residues for substrate recognition. Mutation of arginine 306 to threonine in helix 8 resulted in strongly reduced transport activity. Concomitantly, saturable transport was transformed into a nonsaturable dependence of transport activity on lactate concentration, suggesting that binding of the substrate was strongly impaired. Furthermore, proton translocation in the mutant was partially uncoupled from monocarboxylate transport. Mutation of phenylalanine 360 to cysteine in helix 10 resulted in an altered substrate side chain recognition. In contrast to the wild type transporter, monocarboxylates with more bulky and polar side chains were recognized by the mutated MCT1. Mutation of selected residues in helix 9 and helix 11 (C336A, H337Q, and E391Q) did not cause alterations of the transport properties of MCT1. It is suggested that substrate binding occurs in the carboxy terminal half of MCT1 and that helices 8 and 10 are involved in the recognition of different parts of the substrate. PMID- 10471311 TI - Uno, nessuno e centomila: searching for the identity of mesodermal progenitors. PMID- 10471312 TI - Different effects of bone morphogenetic proteins 2, 4, 12, and 13 on the expression of cartilage and bone markers in the MC615 chondrocyte cell line. AB - In order to study the lineage leading to chondrocyte and osteoblast phenotype in vertebrate development, we examined the effect of recombinant human bone morphogenetic protein (BMP)-2, BMP-4, BMP-12 [or growth and differentiation factor (GDF)-7], and BMP-13 (or GDF-6) on the phenotypic expression of the mouse chondrocyte cell line MC615, grown for 1 or 2 weeks in monolayer. Protein synthesis rates were monitored after incubation with [(14)C]proline. BMP-2 and BMP-4 increased protein synthesis, in agreement with our observation by phase contrast microscopy of a highly refractile matrix around MC615 cells treated with BMP-2 and -4. Markers of the chondrocytic and osteoblastic differentiation were analyzed at mRNA level. Expression of the type II collagen gene, a marker of the cartilage phenotype, was up-regulated in the presence of low concentration of BMP 2 or -4 (50 ng/ml) and down-regulated at higher concentrations (100-400 ng/ml). In parallel, this expression was stable in the presence of BMP-12 or -13 in the dose range tested (50-400 ng/ml). Expression of the matrix Gla protein (MGP) gene, another marker of cartilage, was also reduced in the presence of 100 ng/ml BMP-2 or -4, while it remained stable in the presence of BMP-12 or -13 at the same concentration. In contrast, expression of the bone Gla protein (BGP) gene, or osteocalcin, a marker of the bone phenotype, was induced when the cells were treated with BMP-2 or -4 but was not detected when the cells were treated with BMP-12 or -13. At the same time, BMP-2 or -4 markedly up-regulated expression of type X collagen mRNA, indicating that MC615 cells possess the ability to express traits associated with endochondral ossification, when exposed to specific BMPs. Furthermore, detailed analysis of type II collagen expression showed that the alternatively spliced transcript collagen IIB, specific for cartilage, is expressed concomitantly with BGP. Therefore, MC615 chondrocytes can simultaneously express chondrocytic and osteoblastic markers, in response to BMP 2 or -4, but show minimal response to BMP-12 (or GDF-7) or to BMP-13 (or GDF-6). These results raise the possibility that chondrocytes in vivo can express osteoblastic properties, provided they are induced by BMP-2 or -4. PMID- 10471313 TI - Skeletal unloading induces biphasic changes in insulin-like growth factor-I mRNA levels and osteoblast activity. AB - To determine the local mechanisms involved in the effects of skeletal unloading on bone formation, we studied the temporal pattern of mRNA levels for insulin like growth factor-I (IGF-I), IGF-I receptor type I (IGF-IR), and transforming growth factor beta receptor type II (TGF-betaRII) in relation to osteoblast phenotypic markers and osteoblast activity in hindlimb suspended rats. Skeletal unloading decreased bone volume and the mineralizing and osteoblastic surfaces at 4, 7, and 14 days in the tibial metaphysis, whereas the mineral appositional rate returned to normal at 14 days of suspension. RT-PCR analysis showed that skeletal unloading decreased type 1 collagen (Col 1) and osteocalcin (OC) mRNA levels in metaphyseal bone at days 4 and 7, and the levels returned to normal at 14 days of suspension. Unloading also decreased mRNA levels for IGF-I, IGF-IR, and TGF betaRII at 4-7 days in the metaphyseal bone. However, IGF-I and IGF-IR levels rose above normal at 14 days of suspension. The biphasic changes in IGF-I mRNA levels were strongly correlated with Col 1 and OC mRNA levels. The associated biphasic pattern of IGF-I/IGF-IR expression, osteoblast markers, and osteoblast activity strongly suggests an important role for IGF-I signaling in the local effect of skeletal unloading on metaphyseal bone formation. PMID- 10471314 TI - Stimulated phosphorylation of intracellular connexin43. AB - A monoclonal antibody, Zymed 13-8300, was previously reported to only detect nonphosphorylated connexin43 (Nagy et al., Exp. Cell Res. 236, 127-136, 1997). We show that 13-8300 can detect several phosphorylated species of connexin43 in Western blots after stimulation of two fibroblast cell systems with fresh growth medium, 12-O-tetradecanoyl phorbol-13-acetate, pervanadate, or permolybdate. In one of the cell systems, at least three forms of phosphorylated connexin43 could migrate at the same position during electrophoresis. The comigration of differentially phosphorylated species may complicate the molecular and functional analysis of phosphorylation sites in Cx43. Immunofluorescence experiments indicated that the newly generated phosphorylated Cx43 forms mainly had a perinuclear location. Also, in cells treated with brefeldin A for 8 h, in which the majority of connexin43 was intracellular, phosphorylation was induced by the agents. Phosphorylation of intracellular connexin43 can therefore be induced by several stimuli. PMID- 10471315 TI - Analysis of nuclear localization signals using a green fluorescent protein-fusion protein library. AB - We describe here an efficient method for identifying intracellular localization signals in proteins with stereospecific intracellular localizations in culture cells. The method involves rapid fluorescence screening of cells transfected with a cDNA library in which cDNAs are fused to the gene encoding the Aequorea victoria green fluorescent protein (GFP). We analyzed nuclear localization and nuclear localization signals (NLSs) in a model application of this method. As a result, we identified classical NLSs in 75% of nuclear localized proteins. We identified some novel NLS candidates among the classical NLS-negative sequences whose nuclear localization was also identified in another cell line and with other molecular tag sequences. This method will be useful for identifying intracellular localization signals and for more detailed analysis of intracellular architecture. PMID- 10471316 TI - Role of hyperpolarization attained by linoleic acid in chick myoblast fusion. AB - Our previous report has suggested that hyperpolarization generated by reciprocal activation of calcium-activated potassium (K(Ca)) channels and stretch-activated channels induces calcium influx that triggers myoblast fusion. Here we show that linoleic acid is involved in the process of generating hyperpolarization in cultured chick myoblasts and hence in promotion of the cell fusion. Linoleic acid dramatically hyperpolarized the membrane potential from -14 +/- 3 to -58 +/- 5 mV within 10 min. This effect was partially blocked by 1 mM tetraethylammonium (TEA) or 30 nM charybdotoxin, a selective K(Ca) channel inhibitor, and completely abolished by 10 mM TEA. Single-channel recordings revealed that linoleic acid activates TEA-resistant potassium channels as well as K(Ca) channels. Furthermore, linoleic acid induced calcium influx from extracellular solution, and this effect was partially blocked by 1 mM TEA and completely prevented at 10 mM, similar to the effect of TEA on linoleic acid-mediated hyperpolarization. Since the valinomycin-mediated hyperpolarization promoted calcium influx, hyperpolarization itself appears capable of inducing calcium influx. In addition, gadolinium prevented the valinomycin-mediated increase in intracellular calcium level under hypotonic conditions, revealing the involvement of stretch-activated channels in calcium influx. Furthermore, linoleic acid stimulated myoblast fusion, and this stimulatory effect could completely be prevented by 10 mM TEA. These results suggest that linoleic acid induces hyperpolarization of membrane potential by activation of potassium channels, which induces calcium influx through stretch-activated channels, and thereby triggers myoblast fusion. PMID- 10471317 TI - Engineered smooth muscle tissues: regulating cell phenotype with the scaffold. AB - Culturing cells on three-dimensional, biodegradable scaffolds may create tissues suitable either for reconstructive surgery applications or as novel in vitro model systems. In this study, we have tested the hypothesis that the phenotype of smooth muscle cells (SMCs) in three-dimensional, engineered tissues is regulated by the chemistry of the scaffold material. Specifically, we have directly compared cell growth and patterns of extracellular matrix (ECM) (e.g. , elastin and collagen) gene expression on two types of synthetic polymer scaffolds and type I collagen scaffolds. The growth rates of SMCs on the synthetic polymer scaffolds were significantly higher than on type I collagen sponges. The rate of elastin production by SMCs on polyglycolic acid (PGA) scaffolds was 3.5 +/- 1.1 fold higher than that on type I collagen sponges on Day 11 of culture. In contrast, the collagen production rate on type I collagen sponges was 3.3 +/- 1.1 fold higher than that on PGA scaffolds. This scaffold-dependent switching between elastin and collagen gene expression was confirmed by Northern blot analysis. The finding that the scaffold chemistry regulates the phenotype of SMCs independent of the scaffold physical form was confirmed by culturing SMCs on two-dimensional films of the scaffold materials. It is likely that cells adhere to these scaffolds via different ligands, as the major protein adsorbed from the serum onto synthetic polymers was vitronectin, whereas fibronectin and vitronectin were present at high density on type I collagen sponges. In summary, this study demonstrates that three-dimensional smooth muscle-like tissues can be created by culturing SMCs on three-dimensional scaffolds, and that the phenotype of the SMCs is strongly regulated by the scaffold chemistry. These engineered tissues provide novel, three-dimensional models to study cellular interaction with ECM in vitro. PMID- 10471318 TI - Sequence determinants of nuclear localization in the alpha and beta isoforms of human topoisomerase II. AB - The alpha and beta isoforms of DNA topoisomerase II (topo II) are targets for several widely used chemotherapeutic agents, and resistance to some of these drugs may be associated with reduced nuclear localization of the alpha isoform. Human topo IIalpha contains a strong bipartite nuclear localization signal (NLS) sequence between amino acids 1454 and 1497 (alphaNLS(1454-1497)). In the present study, we show that human topo IIalpha tagged with green fluorescence protein is still detectable in the nucleus when alphaNLS(1454-1497) has been disrupted. Seven additional regions in topo IIalpha containing overlapping potential bipartite NLSs were evaluated for their nuclear targeting abilities using a beta galactosidase reporter system. A moderately functional NLS was identified between amino acids 1259 and 1296. When human topo IIbeta was examined in a similar fashion, it was found to contain two strongly functional sequences betaNLS(1522 1548) and betaNLS(1538-1573) in the region of topo IIbeta comparable to the region in topo IIalpha that contains the strongly functional alphaNLS(1454-1497). The third, betaNLS(1294-1332), although weaker than the other two beta sequences, is significantly stronger than the analogous alphaNLS(1259-1296). Differences in the NLS sequences of human topo II isoforms may contribute to their differences in subnuclear localization. PMID- 10471319 TI - PI3-kinase is involved in mitogenic signaling by the oncogenic receptor tyrosine kinase Xiphophorus melanoma receptor kinase in fish melanoma. AB - Overexpression of the mutationally activated receptor tyrosine kinase Xiphophorus melanoma receptor kinase (Xmrk) initiates formation of hereditary malignant melanoma in the fish Xiphophorus. In melanoma as well as in a melanoma-derived cell line (PSM) this receptor is highly activated resulting in constitutive Xmrk mediated mitogenic signaling. In order to analyze mitogenic signaling triggered by Xmrk a possible involvement of phosphatidylinositol 3 (PI3)-kinase in Xmrk signal transduction was examined. Constitutive binding of the p85 adapter subunit of PI3-kinase to the Xmrk receptor was detected in PSM melanoma cells. Further analyses in BHK cells expressing a Xmrk chimera (HER-mrk) showed that p85 association with the intracellular part of Xmrk was dependent on autophosphorylation of the receptor. In vitro binding studies revealed that the interaction is mediated mainly through the N-terminal SH2 domain of p85 which directly binds to a sequence motif around phosphorylated Tyr-983 in the Xmrk carboxy-terminus. In accordance with recruitment of p85 by Xmrk in PSM cells, the PI3-kinase downstream target Akt was found to be highly phosphorylated on Ser 473, indicating efficient PI3-kinase signaling in melanoma cells. PI3-kinase activation was also detected in Xiphophorus melanoma. Moreover, malignant melanomas exhibited an increased level of PI3-kinase activity which was about three times higher than that in benign pigmented lesions. Inhibition of PI3 kinase activity in PSM melanoma cells by both Wortmannin and LY294002 blocked entry into S-phase. Together these data demonstrate that PI3-kinase is a substrate of the oncogenic Xmrk receptor and plays a significant role in mitogenic signaling of melanoma cells and the formation of malignant melanoma in Xiphophorus. PMID- 10471320 TI - Abnormal sperm parameters in humans are indicative of an abortive apoptotic mechanism linked to the Fas-mediated pathway. AB - The life cycle of many cell types can hinge on the presence of death factors that can control programmed cell death. The Fas-mediated apoptotic pathway has been implicated in controlling apoptosis during spermatogenesis in a number of mammalian species. In the human, the presence of nuclear DNA damage in ejaculated spermatozoa has pointed to a possible role for apoptosis during spermatogenesis. The presence of other molecular markers of apoptosis has, however, not been shown. More importantly, differences in these markers have not been investigated in men with normal and abnormal sperm parameters. In this study we examine for the presence of the cell surface protein Fas in ejaculated human spermatozoa. Ejaculated spermatozoa (55 samples) were labeled with anti-human Fas antibody and the number of spermatozoa displaying Fas were counted using a fluorescence activated cell sorter (FACS). In 30/31 (96.8%) normal males (>20 million sperm per milliliter), less than 10% of the spermatozoa were Fas positive. In contrast, 14/24 (58.3%) oligozoospermic samples (<20 million sperm per milliliter) contained more than 10% Fas-positive spermatozoa. Similar differences were observed in men whose spermatozoa had poor motility and morphology. These results indicate that apoptosis is a major mechanism in regulating spermatogenesis in the human and that there are clear differences in molecular markers of apoptosis between males with normal and abnormal sperm parameters. We propose that the presence of Fas-labeled spermatozoa in the ejaculate of these men is indicative of an "abortive apoptosis" having taken place, whereby the normal apoptotic mechanisms have misfunctioned, have been overridden, or have not been completed. PMID- 10471321 TI - The tetraspanin CD9 influences the adhesion, spreading, and pericellular fibronectin matrix assembly of Chinese hamster ovary cells on human plasma fibronectin. AB - The role of CD9 in cell adhesion and spreading on adhesive proteins was investigated using a transfected Chinese hamster ovary (CHO) cell system. CD9 cell surface expression resulted in reduced adhesion and increased spreading on fibronectin (Fn). Whereas mock-transfected (mock CHO) and naive CHO cells assumed a typical fibroblast spindle shape morphology, CD9-transfected (CD9-CHO) cells were polygonal with many filipodial projections and exhibited a twofold greater surface area. The spread morphology of CD9-CHO cells, but not mock CHO cells, was inhibited by PB1 mAb blockade of alpha(5)beta(1), suggesting that the coexpression of alpha(5)beta(1) and CD9 influenced cell activity on Fn. The second extracellular loop of CD9 was implicated in regulation of adhesion since reduced CD9-CHO cell adhesion on Fn was reversed by either anti-CD9 antibody ligation to the second extracellular loop or with cells expressing a CD9 mutant lacking the second extracellular loop domain. Using cell adhesion assays and ELISA, we demonstrated CD9 binding to the HEP2/IIICS region of Fn. Finally, CD9 expression resulted in a twofold reduction in Fn-rich pericellular matrix assembly. Our observations show that CD9 dramatically influences CHO cell interactions with Fn and suggest that CD9 has an important role in modulating cell-extracellular matrix interactions. PMID- 10471322 TI - Preferential perinuclear localization of poly(ADP-ribose) glycohydrolase. AB - The transient nature of poly(ADP-ribosyl)ation, a posttranslational modification of nuclear proteins, is achieved by the enzyme poly(ADP-ribose) glycohydrolase (PARG) which hydrolyzes the poly(ADP-ribose) polymer into free ADP-ribose residues. To investigate the molecular size and localization of PARG, we developed a specific polyclonal antibody directed against the bovine PARG carboxy terminal region. We found that PARG purified from bovine thymus was recognized as a 59-kDa protein, while Western blot analysis of total cell extracts revealed the presence of a unique 110-kDa protein. This 110-kDa PARG was mostly found in postnuclear extracts, whereas it was barely detectable in the nuclear fractions of COS7 cells. Further analysis by immunofluorescence revealed a cytoplasmic perinuclear distribution of PARG in COS7 cells overexpressing the bovine PARG cDNA. These results provide direct evidence that PARG is primarily a cytoplasmic enzyme and suggest that a very low amount of intranuclear PARG is required for poly(ADP-ribose) turnover. PMID- 10471323 TI - Effects of jarastatin, a novel snake venom disintegrin, on neutrophil migration and actin cytoskeleton dynamics. AB - A new disintegrin, an RGD-containing peptide of 6 kDa called jarastatin, was purified from Bothrops jararaca venom. It is a potent inhibitor of platelet aggregation induced by ADP, collagen, and thrombin. The effect of jarastatin on neutrophil migration in vivo and in vitro and on the actin cytoskeleton dynamics of these cells was investigated. Incubation in vitro with jarastatin significantly inhibited, in a concentration-dependent manner, the chemotaxis of human neutrophils toward fMLP, IL-8, and jarastatin itself. Despite this inhibitory effect, jarastatin induced neutrophil chemotaxis. A significant increase of F-actin content was observed in jarastatin-treated neutrophils. Furthermore, as demonstrated by confocal microscopy after FITC-phalloidin labeling, these cells accumulated F-actin at the plasmalemma, a distribution similar to that observed in fMLP-stimulated cells. Pretreatment of mice with jarastatin inhibited neutrophil migration into peritoneal cavities induced by carrageenan injection. The results suggest that binding of jarastatin to neutrophil integrins promotes cellular activation and triggers a dynamic alteration of the actin filament system and that this is one of the first event in integrin-mediated signaling. PMID- 10471324 TI - Identification of (CAG)(n) and (CGG)(n) repeat-binding proteins, CAGERs expressed in mature neurons of the mouse brain. AB - The trinucleotide repeats (CAG)(n) and (CGG)(n) have been shown to be expanded in responsible genes of several human hereditary neurological disorders. In studies of mice, we previously identified two homologous single-stranded (ss)(CAG) and ss(CGG) repeat-binding proteins, CAGER-1 (44 kDa) and CAGER-2 (40 kDa) (CAG element-recognizing proteins). The specific binding activities of these proteins were predominantly detected in the mouse brain. We have isolated the cDNAs encoding CAGER-1 and CAGER-2 and found that they were identical to previously reported cDNAs for Puralpha and Purbeta, respectively. Puralpha of 28 kDa was previously identified as a replication-origin-binding protein that is ubiquitously expressed in proliferating cells. We show that the transcripts of CAGERs increase after birth and are detected at high levels in the adult mouse brain but at very low or virtually undetectable levels in other mouse tissues. Biochemical properties and molecular weights are different between CAGERs and Puralpha/beta. Immunostaining with specific antibodies against CAGERs indicates that CAGERs in the mouse brain reside in nonproliferating neurons but not in proliferating glia. We conclude that CAGERs and Puralpha/beta are unrelated proteins, and CAGERs are neuronal single-stranded sequence-binding proteins in the mouse brain. Misassignment of cDNAs is described. PMID- 10471325 TI - Poly(ADP-ribose) polymerase inhibition in oxidant-stressed endothelial cells prevents oncosis and permits caspase activation and apoptosis. AB - Endothelial cells (EC) are subject to oxidative-induced cell death. Activation of poly(ADP-ribose) polymerase (PARP) occurs early in oxidant-induced EC injury and putatively mediates cell death by depleting its substrate, NAD(+). In this study, the role of PARP in H(2)O(2)-induced EC death was investigated. EC were exposed to oxidant stress and viability continuously monitored using fluorescent dye exclusion. Inhibition of PARP with 1, 5-dihydroxyisoquinoline (DIQ) delayed the time course of oxidant-induced EC death. Concurrent addition of the protein synthesis inhibitor, cycloheximide, or the endonuclease inhibitor, aurintricarboxylic acid, to PARP-inhibited cells further delayed the onset and attenuated the extent of H(2)O(2)-induced cell lysis, consistent with an active mode of cell death. Caspase-3-like activity, a hallmark of apoptosis, was negligible in oxidant-treated EC alone, however, inhibition of PARP by 3 aminobenzamide or DIQ dramatically increased caspase-3-like activity. Morphological assessment confirmed that the primary mode of death in oxidant stressed EC was oncosis. However, following PARP inhibition, the cells switched to apoptosis. Since inflammation is associated with oncosis and not apoptosis, the results presented here could explain the beneficial effects seen with PARP inhibition in various in vivo models of oxidant injury and provide a mechanism to manipulate this injury into a state of cell death that could ultimately be controlled. PMID- 10471326 TI - E1A oncogene induction of cellular susceptibility to killing by cytolytic lymphocytes through target cell sensitization to apoptotic injury. AB - E1A oncogene expression increases mammalian cell susceptibility to lysis by cytolytic lymphocytes (CLs) at a stage in this intercellular interaction that is independent of cell surface recognition events. Since CLs can induce either apoptotic or necrotic cell death, we asked whether E1A sensitization to injury induced apoptosis is sufficient to explain E1A-induced cytolytic susceptibility. Mouse, rat, hamster, and human cells that were rendered cytolytic susceptible by E1A were also sensitized to CL-induced and chemically induced apoptosis. In contrast, E1A-positive cells were no more susceptible to injury-induced necrosis than E1A-negative cells. Similar to induction of cytolytic susceptibility and in contrast to other E1A activities, cellular sensitization to chemically induced apoptosis depended on high-level E1A oncoprotein expression. Loss of both cytolytic susceptibility and sensitization to chemically induced apoptosis was coselected during in vivo selection of E1A-positive sarcoma cells for increased tumorigenicity. Furthermore, E1A mutant proteins that cannot bind the cellular transcriptional coactivator, p300, and that fail to induce cytolytic susceptibility also failed to sensitize cells to injury-induced apoptosis. These data indicate that E1A induces susceptibility to killer cell-induced lysis through sensitization of cells to injury-induced apoptosis. PMID- 10471327 TI - Coordinated regulation of radioadaptive response by protein kinase C and p38 mitogen-activated protein kinase. AB - Eukaryotic cells are known to have an inducible or adaptive response that enhances radioresistance after a low priming dose of radiation. This radioadaptive response seems to present a novel cellular defense mechanism. However, its molecular processing and signaling mechanisms are largely unknown. Here, we studied the role of protein kinase C (PKC) and mitogen-activated protein kinase (MAPK) in the expression of radioadaptive response in cultured mouse cells. Protein immunoblot analysis using isoform-specific antibodies showed an immediate activation of PKC-alpha upon X-irradiation as indicated by a translocation from cytosol to membrane. A low priming dose caused a prolonged translocation, while a nonadaptive high dose dramatically downregulated the total PKC level. Low-dose X-rays also activated the p38 MAPK. The activation of p38 MAPK and resistance to chromosome aberration formation were blocked by SB203580, an inhibitor of p38 MAPK, and Calphostin C, an inhibitor of PKC. Furthermore, it was demonstrated that p38 MAPK was physically associated with delta1 isoform of phospholipase C (PLC-delta1), which hydrolyzed phosphatidylinositol bisphosphate into diacylglycerol, an activator of PKC, and that SB203580 also blocked the activation of PKC-alpha. These results indicate the presence of a novel mechanism for coordinated regulation of adaptive response to low-dose X-rays by a nexus of PKC-alpha/p38 MAPK/PLC-delta1 circuitry feedback signaling pathway with its breakage operated by downregulation of labile PKC-alpha at high doses or excess stimuli. PMID- 10471328 TI - Characterization of the soluble form of the low density lipoprotein receptor related protein (LRP). AB - We report characterization of the soluble form of the low density lipoprotein receptor-related protein (sLRP) which circulates in human plasma. Amino acid sequence analysis confirmed that sLRP isolated from human plasma contains the alpha-chain of LRP1. In addition, Western blot analysis identified a truncated beta-chain noncovalently associated with the purified alpha-chain. The molecular size (M(r) 55K) of the peptide portion of the truncated beta-chain indicates that the subunit comprises the extracellular portion of the beta-chain and terminates in a membrane-proximal region. We investigated the mechanism by which sLRP may be generated using the trophoblast cell line, BeWo, which releases sLRP in culture. Cell surface labeling experiments indicate that LRP is released from BeWo cells following expression at the cell surface. Incubation of BeWo cells in the presence of a metalloproteinase inhibitor, INH-3855-PI, results in a dose dependent inhibition of LRP shedding. The metalloproteinase responsible for the shedding of LRP by BeWo cells is not up-regulated by phorbol ester and is not dependent on serine proteases, such as plasmin, for activity. The BeWo cell line is derived from a human gestational choriocarcinoma and preliminary studies suggest that LRP may be shed within the placenta during gestation. Increased levels of sLRP were detected in cord blood. In term placenta, LRP is expressed in the syncytium, which comprises the maternal-fetal interface. Increased levels of sLRP in cord blood may reflect cellular dysfunction and increased metalloproteinase activity at this important interface. PMID- 10471329 TI - Regulation and deregulation of E2F1 in postmitotic neurons differentiated from embryonal carcinoma P19 cells. AB - Neurons withdraw from the cell cycle immediately after differentiation from their proliferative precursors. E2F1, a principal transcription factor that promotes cell cycle progression, must be silenced in neurons. We investigated the E2F1 system in postmitotic neurons derived from murine embryonal carcinoma P19 cells. P19 cells highly expressed the E2F1 gene during neural differentiation, and enriched neurons contained a high abundance of E2F1 mRNA. In contrast, postmitotic neurons possessed extremely low levels of E2F1 protein as assessed by the electrophoretic mobility shift assay and Western blotting. A recombinant E2F1 fusion protein was ubiquitinated in vitro when incubated with neuronal lysates. In addition, treatment with the proteasome inhibitor MG132 increased the endogenous level of E2F1 protein in neurons. These results suggest that the ubiquitin-proteasome pathway contributes, at least in part, to the downregulation of E2F1 protein in postmitotic neurons. Adenovirus-mediated transfer of E2F1 cDNA into postmitotic neurons induced both bromodeoxyuridine incorporation and chromatin condensation, suggesting that deregulated E2F1 expression causes both aberrant S-phase entry and apoptosis of postmitotic neurons. Thus, downregulation of endogenous E2F1 protein in postmitotic neurons may be indispensable for the prevention of their reentry into the cell cycle. PMID- 10471330 TI - Effects of anti-PM-Scl 100 (Rrp6p exonuclease) antibodies on prenucleolar body dynamics at the end of mitosis. AB - Prenucleolar bodies (PNBs) are transitory structures which serve as building blocks for nucleoli at the transition mitosis/interphase. The assembly of PNBs and their pathway are not clearly understood. To better understand these events, the behavior of the PNB-containing PM-Scl 100 protein was compared with that of other PNB proteins. This nucleolar protein was chosen because its yeast homologue, Rrp6p exonuclease [1], is known to participate in late events in 5.8 S rRNA (ribosomal RNA) processing. There was a heterogeneous distribution of nucleolar proteins in different classes of PNBs. The PM-Scl 100 colocalized predominantly with protein B23. The PM-Scl-100-containing PNBs were translocated at later times to nucleoli as opposed to the fibrillarin-containing PNBs. Microinjections of antibodies directed against PM-Scl 100 during mitosis inhibited targeting of PM-Scl 100 to the nucleolus. However fibrillarin and protein B23 still participated in nucleolar assembly in early G1. We conclude that there are different kinds of PNBs whose translocation to the nucleoli follow ordered kinetics. Interestingly, proteins involved in late steps of processing as PM-Scl 100 are translocated late, suggesting that they are not cotranscriptionally associated with the rRNA precursors. PMID- 10471331 TI - Cytokine signal transduction in P19 embryonal carcinoma cells: regulation of Stat3-mediated transactivation occurs independently of p21ras-Erk signaling. AB - Ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF) are members of a subfamily of related cytokines that share gp130 as common signal transducing receptor component. CNTF has recently been demonstrated to induce increased survival and neuronal differentiation of P19 embryonal carcinoma (EC) cells; however, the molecular mechanisms underlying these effects are still elusive. Here we report that CNTF and LIF, but not interleukin-6, activated signal transducers and activators of transcription (STAT)-reporter constructs in P19 EC cells. Supershift analysis revealed that the STAT-element binding complex contained the transcription factor Stat3. Binding of Stat3 was inhibited by protein tyrosine kinase inhibitors, but not by the broad serine/threonine protein kinase inhibitor, H7. However, H7 inhibited CNTF-induced Stat3 transactivation. Using a dominant-negative p21ras construct and a specific inhibitor of mitogen activated protein kinase kinase (MEK; PD098059) we demonstrated that CNTF-induced Stat3 transactivation was independent of the p21ras-mitogen-activated protein kinase (MAPK) pathway, while CNTF-induced MAPK activation was p21ras- and MEK dependent. Taken together, our results demonstrate the activation of the p21ras MAPK and STAT signal transduction pathways in response to CNTF and LIF in P19 EC cells and reveal that there is no modulating crosstalk between these pathways. Furthermore, our data suggest that CNTF- and LIF-induced Stat3 activation in P19 EC cells involves an H7-sensitive p21ras/MAPK- and Ca(2+)-independent kinase. PMID- 10471333 TI - Normal human telomeres are not late replicating. AB - Telomeres in yeast are late replicating. Genes placed next to telomeres in yeast can be repressed (telomere positional effects), leading to the hypothesis that telomeres may be heterochromatic and may control the expression of subtelomeric genes. In addition, yeast telomeres are processed to have a transient long overhang at the end of S phase. The applicability of the yeast data to human biology was examined by determining the timing of telomere replication and processing in normal human diploid fibroblasts. Telomeres were purified from synchronized cells that had been labeled with 5-bromodeoxyuridine (BrdU) at hourly intervals, and the fraction of labeled telomeres was analyzed by retrieval with anti-BrdU antibodies. We determined that normal human telomeres replicate throughout S phase rather than being very late replicating. Furthermore, the overall timing of replication was unaffected by telomere length in young versus old cells or cells whose telomeres had been elongated following transfection with the catalytic subunit of telomerase. Finally, the asymmetry in the length of the G-rich overhang in daughter telomeres produced by leading versus lagging strand synthesis was shown to be established within 1 h of telomere replication, indicating there is no significant delay between synthesis and the processing events that contribute to the establishment of asymmetric overhangs. Therefore, the timings of replication and processing of human telomeres are very different from those of yeast. PMID- 10471332 TI - Extensive clear zone and defective ruffled border formation in osteoclasts of osteopetrotic (ia/ia) rats: implications for secretory function. AB - The cellular distribution of osteoclast integrin subunits alpha(v) and beta(3), the tissue distribution, and level of the apparent ligand osteopontin (OPN) as well as of the putative regulatory enzyme tartrate-resistant acid phosphatase (TRAP) were studied along with the intracellular distribution of the activation marker c-src in osteopetrotic ia/ia (incisors-absent) mutant rats and their normal littermates. In ia/ia rats, the osteoclasts are incapable of bone matrix resorption. Ultrastructurally the cells exhibit extended clear zones at the expense of ordinary ruffled borders. A secretory dysfunction in the mutant is strongly suggested by the absence of detectable extracellular TRAP, concomitant with an accumulation of the enzyme in abundant small cytoplasmic vesicles. Moreover, TRAP mRNA, protein content, as well as enzymatic activity were elevated. Furthermore, increased levels of integrin subunits alpha(v) and beta(3) were detected at the clear zone of mutant osteoclasts. OPN mRNA levels were elevated in long bones from mutants. In ia/ia rats, immunolabeling for OPN was homogeneously distributed at the surface facing osteoclasts, while in normal littermates it was concentrated at the clear zone area and barely detectable at ruffled borders. The absence of OPN labeling in the abundant, putative intracellular secretory vesicles in mutant osteoclasts suggests that these cells do not produce OPN. The osteoclasts of ia/ia rats appeared to produce and translocate the c-src protein to the cell membrane. In ia/ia a defect ruffled border-formation is observed along with extensive clear zone formation and decreased secretory function. The lesion may be due to a signaling defect, but in that case the defect seems to be located downstream to or not involving the c-src pathway. Our results illustrate the close relationship between secretory function and ruffled border formation in osteoclasts, a relationship that appears to be necessary for proper resorptive function. PMID- 10471334 TI - Stimulus-specific and cell type-specific cascades: emerging principles relating to control of apoptosis in the eye. AB - Apoptosis has a critical role in development, homeostasis, wound healing, and the pathophysiology of disease in the organs of multicellular organisms. It has been implicated in these processes in retina, lens, cornea, trabecular meshwork, optic nerve, and the central nervous system pathways that contribute to vision. Considerable interest has been focused on inhibiting apoptosis to control disease and wound healing processes in which programmed cell death is thought to have a critical role. A simplified view led to the search for effective inhibitors of 'the final common pathway for apoptosis'. Recent studies have provided important insights into the modulators that participate in and regulate the apoptosis cascades which are activated in response to cytokines, ionizing radiation, chemotherapeutic agents, growth factor deprivation, and other stimulators of cell death. These studies lead to the inescapable conclusion that the apoptosis pathways are not only stimulus-specific, but also cell-type specific. These observations have important implications related to development of pharmacological strategies for controlling apoptosis-associated disease and apoptosis-initiated wound healing. PMID- 10471335 TI - Fine structure of the developing avian corneal stroma as revealed by quick freeze, deep-etch electron microscopy. AB - Corneal transparency depends on the precise organization of the stromal extracellular matrix. The morphology of the extracellular matrix of the embryonic and adult avian secondary posterior cornea was studied in glutaraldehyde-fixed, quick-frozen, deep-etched replicas with the electron microscope. Although the collagen fibrils changed from a loose network to a more ordered parallel lamellar arrangement during development, their mean diameter remained constant between 30.3 and 31.2 nm. Besides collagen fibrils, other extracellular matrix components were observed: (i) straight or Y-shaped cross-bridging interfibrillar 8-10 nm filaments with 18-22 nm globules; (ii) relatively loose networks of 10-20 nm beaded filaments, with a mean periodicity of 107 nm, often running perpendicular to the collagen fibrils and adhering to the plasma membrane of stromal cells at early developmental stages; (iii) straight or curved 6-12 nm strands forming sheets within the stromal matrix that progressively disappeared, whereas similar structures persisted at the Descemet's membrane-stroma interface; (iv) dense networks of filaments with 6-8 nm filaments, sometimes polygonally arranged, and a substructure of 2-3 nm filaments with globular domains, which progressively disappeared with maturation but remained at the Descemet's membrane-stroma interface; (v) polygonal networks of 9-10 nm filaments with globular domains adherent to the surface of cell plasma membranes at early developmental stages. The temporal expression of deep-etched supramolecular structural assemblies is compatible with that of the so-called 'interstitial basement membrane components' previously described. The quick-freeze and deep-etching method can reveal important aspects of the in situ organization of the corneal extracellular matrix. PMID- 10471336 TI - Modelling cortical cataractogenesis 21: in diabetic rat lenses taurine supplementation partially reduces damage resulting from osmotic compensation leading to osmolyte loss and antioxidant depletion. AB - The concentration of taurine and the amino acids, glutathione, cysteine, ascorbate and ATP were determined in the lenses of rats made diabetic with streptozotocin. In the clear lenses, prior to vacuole formation after 1 or 2 weeks of diabetes, the increase in concentration of sorbitol and the total decrease of all these osmolytes were not significantly different. The major components of the osmolytes lost were taurine and amino acids, which together accounted for over 75% of the total osmolyte loss. Since glutathione, ascorbate, taurine and cysteine have been reported to have antioxidant activity, it appears that their loss may potentiate damage occurring as a result of free radicals generated by nonenzymic glycation by the Maillard reaction. Amino acids also lost as a result of the osmotic compensation, are estimated to be responsible for almost half of the antioxidant activity lost. To test this hypothesis, normal and streptozotocin diabetic female Wistar rats were given taurine at 0.05% or 0.10% (w/w) in the diet. This treatment resulted in small only marginally significant increases in serum taurine levels. At the end of 6 weeks the rats were examined for weight gain or loss and at the time of killing, blood was collected for measurement of serum glucose. gamma-Crystallin levels were determined in vitreous and aqueous humours using a radioimmunoassay. A lens from each rat was homogenized in 8 m guanidinium chloride for adenosine triphosphate (ATP) analysis. In normal rats, a small amount of gamma-crystallin was found in the vitreous humour, and an even smaller amount in the aqueous humour. Diabetes caused a 4- to 5-fold increase in the vitreous humour and a 4-fold increase in gamma-crystallin in the aqueous humour. Diabetes also led to a significant worsening in general body condition, loss of body weight, formation of cataracts, and decrease in lens ATP levels. Addition of taurine to the diet of diabetic animals resulted in a significant decrease of gamma-crystallin leakage into the vitreous but not the aqueous humour. Taurine had no effect on the lens ATP levels. Neither streptozotocin diabetes nor taurine in the diet appeared to affect the weight of the lenses. PMID- 10471337 TI - Modelling cortical cataractogenesis 22: is in vitro reduction of damage in model diabetic rat cataract by taurine due to its antioxidant activity? AB - The protective effect of taurine in model in vitro diabetic cataract and the mechanism of this effect were investigated in isolated rat lenses. Isolated rat lenses were incubated in medium 199 in elevated glucose (55.6 m m) with taurine (5 m m). Taurine concentrations in the lenses were determined by amino acid analysis. Accumulative leakage of the intracellular enzyme lactate dehydrogenase (LDH) was used to estimate damage to the lens, as previously reported. In the clear lenses, prior to vacuole formation, after 1 or 2 days of incubation, the taurine and amino acids in lenses decreased progressively in concentration. In lenses incubated with 5 m m taurine, the level of taurine was increased towards that of control lenses. In taurine-treated lenses LDH leakage was significantly decreased, and lens clarity was maintained, similarly to that found previously for vitamin C and lipoic acid. To test whether taurine has similar antioxidant activity, we tested its ability to decrease luminol luminescence generated by (1) superoxide from hypoxanthine/xanthine oxidase and (2) peroxide from diluted glucose/glucose oxidase. For either superoxide or peroxide, the luminescence was decreased to zero, as a function of increasing taurine concentration, at 30 m m, approximately the physiological concentration of taurine in the lens. Spin trapping confirmed that taurine scavenged superoxide. This is consistent with a role for taurine as an important antioxidant protecting the lens against oxidative insults. Amino acids also had antioxidant activity in this assay, and as a group, when all activities were summed, their loss also contributed significantly to the antioxidant loss. Taken in conjunction with Wolff and Crabbe's observation of increased free radical generation by glucose auto oxidation in diabetes, this suggests a push-pull mechanism for increased oxidative stress in diabetic cataract, involving both increased free radicals and decreased radical scavenging antioxidants. PMID- 10471338 TI - Effects of elevated intraocular pressure on haemoglobin oxygenation in the rabbit optic nerve head: a microendoscopical study. AB - Intraocular pressure dependent reactions of optic nerve head vasculature and intracapillary haemoglobin oxygenation (HbO(2); oxygen saturation) were studied in the center and at the rim of the rabbit optic nerve head (ONH) as well as in the choroid, by a new combination of microendoscopy and simultaneous haemoglobin spectrophotometry. In 13 anesthetized albino rabbits the vasculature and the intracapillary Hb-oxygenation were studied by a microendoscope which was introduced into the eye bulb. Photometric measurements were performed via a beam splitter with the Erlangen micro-lightguide spectrophotometer (EMPHO) from the center of the endoscopic picture. The haemoglobin oxygenation was calculated by real time analysis of the spectral curves. Intraocular pressure was elevated stepwise from 20-80 mmHg. At the rim of the optic nerve head the vascular diameters as well as the intracapillary HbO(2)-values were stable till an intraocular pressure of 60 mmHg and decrease after IOP elevation to 70 and 80 mmHg. In contrast, in the center of the optic nerve head and in the choroid these parameters decline already from 40-50 mmHg on. At an IOP of 60 mmHg (P<0.01) and 70 mmHg (P<0.05) HbO(2)is significantly lower in the ONH center than at the rim. In the center and the choroid HbO(2)is well maintained between 20 and 40 mmHg. After pressure release at the end of the experiment HbO(2)increased to 94.3+/ 4.6% (rim) and 98.8+/-1.5% (center) of the initial value at 20 mmHg (difference not significant).By the high spatial resolution of this new optical method we were able to demonstrate that the center of the optic nerve head is more sensitive to changes in intraocular pressure than the optic nerve head rim. Thus, tissue damage after critical haemodynamic and oxygenation parameters seems more probable in the relatively poor perfused center of the ONH than in the overperfused rim. PMID- 10471339 TI - Chondroitin sulfate proteoglycan core proteins in the interphotoreceptor matrix: a comparative study using biochemical and immunohistochemical analysis. AB - This study characterizes the core proteins of chondroitin sulfate-type glycosaminoglycans located in the interphotoreceptor matrix and establishes the tissue distribution of chondroitin immunoreactivity in human, bovine, mouse and rat retinas. Monoclonal antibodies specific to unsulfated (DeltaDiOS), 4-sulfated (DeltaDi4S) and 6-sulfated (DeltaDi6S) chondroitin were employed. Retinal sections and IPM samples were either (a) digested with chondroitinase ABC to expose antibody specific epitopes, (b) double digested with chondroitinase ABC and chondroitinase AC II to remove specific epitopes, or (c) left undigested to evaluate mimotope labeling. In tissue sections from each species studied, positive immunoreactivity to the DeltaDi6S antibody was present in the IPM surrounding both rods and cones. In human and bovine, DeltaDi6S labeling of the cone matrix compartments was more intense than labeling of the matrix surrounding rods. Intense DeltaDi6S immunoreactivity was present surrounding the foveal cones. In mouse and rat, no differences in labeling intensity of IPM surrounding rod and cone photoreceptors were evident, although labeling of the IPM near the apical surface of the retinal pigment epithelium and around the photoreceptor inner segments was more pronounced than that surrounding the outer segments. All DeltaDi6S antibody labeling was eliminated with chondroitinase AC II digestion. No IPM immunoreactivity in tissue sections was observed when the DeltaDi0S or DeltaDi4S antibodies were used. In Western blots of IPM extracts treated with chondroitinase ABC, prominent DeltaDi6S immunoreactive bands were present at approximately 230 kD and 150 kD in each species studied, with the exception of the human, where the 150 kD component is not a chondroitin proteoglycan. Each of the prominent DeltaDi6S immunoreactive bands showed minor immunoreactivity to the DeltaDi4S antibody. No DeltaDi0S immunoreactivity was noted in Western blots of IPM samples from any species. All immunoreactivity was lost following chondroitinase AC II digestion. These observations document similarities in the electrophoretic mobility of IPM proteoglycan core proteins released following chondroitinase ABC digestion in the four species studied, but reveal pronounced differences in the tissue distribution. Bovine and human IPM show greater concentrations of DeltaDi6S immunoreactivity surrounding cones than rods, whereas rodent tissues show higher concentrations near the retinal pigment epithelium and around the photoreceptor inner segments than around the outer segments. The pattern of distribution of these proteoglycan molecules is highly conserved in these species, suggesting a common role in IPM structure and function. PMID- 10471340 TI - Ca(2+)-ATPase activity and lens lipid composition in reconstituted systems. AB - Lens lipid composition and lipid hydrocarbon chain structure change with age, region and cataract. Since the lens Ca(2+)-ATPase pump is important to the maintenance of calcium homeostasis and lens clarity, muscle sarcoplasmic reticulum Ca(2+)-ATPase was reconstituted with bovine lens lipids and dihydrosphingomyelin, the rare and major phospholipid of the human lens. Ca(2+) ATPase activity was found to be about 5 times lower when the pump was reconstituted into dihydrosphingomyelin or lens lipids compared to native sarcoplasmic reticulum lipids. The addition of cholesterol to levels ranging from 13-53 mole%, had no affect on reconstituted Ca(2+)-ATPase activity. Ca(2+)-ATPase activity correlated with the degree of hydrocarbon chain saturation. The greater levels of saturation are a consequence of the high sphingolipid content in the reconstituted systems. These data support the hypothesis that changes in lens lipid composition or structure could affect Ca(2+)-ATPase activity in human lenses. Because the mechanisms governing Ca(2+)-ATPase activity in vivo are much more complex than in these simple reconstituted systems, this study represents an initial step in the elucidation of the relationships of endogenous membrane lipid composition-structure and function. PMID- 10471341 TI - Topically applied betaxolol attenuates NMDA-induced toxicity to ganglion cells and the effects of ischaemia to the retina. AB - The present results show that topically applied Betoptic(R)(0.5% betaxolol) to the rabbit or rat eye reaches the retina and can counteract the detrimental effects caused by ischaemia/reperfusion or N -methyl- d -aspartate (NMDA)-induced insults to the retina. Betaxolol is a beta(1)-adrenergic blocker but its neuroprotective action is generally thought to be due to its calcium channel blocking properties. Support for this view comes from studies on cultures of cortical neurones where it was found that betaxolol attenuated the NMDA-induced influx of(45)Ca(2+)while beta-adrenoreceptor agonists were ineffective. Topically applied Betoptic(R)to the rabbit eye was observed to reach the retina in maximal amounts within 60 min. Some of the substance was also found in the contralateral retina of the untreated eye suggesting that the agent reaches the retina by local systemic and retinal circulation. Concurrent treatment with Latanoprost(R)did not result in a greater amount of betaxolol reaching the retina. An ophthalmodynamometric procedure, which raises the intraocular pressure, was used to apply an ischaemic insult to the rabbit retina. After three days of reperfusion the b-wave of the electroretinogram was reduced by an average of 59% and the choline acetyltransferase immunoreactivity in the retina was almost obliterated. However, when experiments were carried out on animals which had been treated with one drop of Betoptic(R) twice daily for 4 weeks before ischaemia and also during the reperfusion phase, the reductions in both the b-wave of the electroretinogram and retinal choline acetyltransferase immunoreactivity due to ischaemia/reperfusion were greatly attenuated. Intravitreal injection of NMDA into the rat eye caused a decrease in the immunostaining for Thy-1 antigen which is associated with ganglion cells. The Thy-1 mRNA level was also reduced as was the mRNA for the common subunit of the NMDA receptor, the NR1 subunit. However, in animals subjected to a topical Betoptic(R)regime, before and after intravitreal injection of NMDA, the decreases in the mRNA levels of Thy-1 and NR1 were significantly attenuated. PMID- 10471342 TI - Organization of the rabbit vitreous body: lamellae, Cloquet's channel and a novel structure, the 'alae canalis Cloqueti'. AB - Even though the rabbit is a frequently used animal model for studies on vitreous function and pathobiology, data on the structural organization of the rabbit vitreous are scarce. The aim of the present study is to give a detailed description of rabbit vitreous structure in order to provide a basis for studies on changes in vitreous organization induced by pathophysiological processes. We studied the vitreous body of adult rabbit eyes by complementary anatomical evaluation methods, by light microscopy and by transmission electron microscopy. Regional and local variations in vitreous matrix organization were observed. Regionally, a cortex, an intermediate area, and a centre were distinguished. In addition, specific structures were locally observed. Lamellae run through the intermediate area in a funnel-like pattern, converging upon the asymmetrically positioned optic disc. A central channel (Cloquet's channel) was found in all eyes. We demonstrated a novel structure, attached to Cloquet's channel and to the medullary rays. Because of its wing-shaped sheet-like morphology, we named it the 'alae canalis Cloqueti'. PMID- 10471343 TI - Hypoxia inhibits the changes in action potentials and ion channels during primary culture of neonatal rat ventricular myocytes. AB - Action potentials of rat ventricular myocytes are progressively shortened after birth within several weeks mainly due to a progressive increase in transient outward potassium current (I(to)). On the supposition that an elevation in blood oxygen after birth may contribute to such developmental change, we studied effects of long-term exposure to hypoxia on changes in cardiac action potentials and I(to). Single ventricular myocytes isolated from day-old neonatal rat hearts were cultured in normoxic condition (21% O(2)) for 15 days and served as control. To test the influence of long-term exposure to hypoxia, O(2)tension was reduced to 7.5% at day 6 during culture. In 15-day cells cultured in normoxia, action potential duration (APD) was shortened by 44% (n=11) compared with 5-day cells (n=10); cell capacitance was increased to 2.0-fold. I(to)density was increased by 189-265% (n=11) at voltage levels from -20 to 50 mV without any changes in the kinetics of current inactivation. In 15-day cells cultured in hypoxia, APD was shortened only by 16% (n=6) from control; the increment of cell capacitance was 2.1-fold (n=6). The I(to)increment was limited to 53% (n=8); both inactivation and its recovery of the current was apparently slowed due to the amplification of the slower component. These results suggest that the developmental augmentation of I(to)expression during culture requires oxygen and the increase in I(to)and cell hypertrophy are likely regulated independently. PMID- 10471344 TI - Mechanism of SNAP potentiating antiproliferative effect of calcitonin gene related peptide in cultured vascular smooth muscle cells. AB - We previously showed that CGRP inhibits cell proliferation which correlates with an elevation of cAMP levels in rabbit aortic vascular smooth muscle cells (VSMCs). The present study determined the effects of S-nitroso-N acetylpenicillamine (SNAP, a nitric oxide donor) on CGRP-induced antiproliferative effects and cellular mechanism in cultured rabbit aortic VSMCs. The cells (in fifth-sixth passage) were exposed to 2.5% fetal bovine serum for 24 h in the presence or absence of SNAP, hCGRP or both.(3)H-thymidine incorporation was used to measure DNA synthesis. The results showed that SNAP (60-100 microm) significantly inhibited the proliferation and elevated cGMP levels in cultured rabbit aortic VSMCs. In combination, however, SNAP (30 microm) potentiated hCGRP (10-100 n m)-induced antiproliferation. SNAP (30 microm) and hCGRP (10-100 n m) or forskolin (10 microm), an activator of adenylate cyclase, caused more than additive cAMP elevations, but not cGMP elevations, in these cells. Quazinone, an inhibitor of cGMP-inhibited-phosphodiesterase (cGI-PDE, PDE3), or SNAP plus quazinone caused a similar potentiation as SNAP of the hCGRP-induced elevations of cAMP levels. The data indicate that SNAP-induced potentiation of CGRP's effects likely involves inhibition of cGI-PDE, thus allowing enhanced accumulation of cAMP that mediates the antiproliferative effects of hCGRP in cultured rabbit aortic VSMCs. PMID- 10471345 TI - Extracellular matrix regulation in the development of Syrian cardiomyopathic Bio 14.6 and Bio 53.58 hamsters. AB - The myocardium contains a collagen matrix composed primarily of collagen and fibronectin, which are major determinants of the myocardial architecture, structural integrity and mechanical properties. The present study was undertaken to determine the age-related changes of the accumulation and degradation of the collagen matrix in Syrian myopathic hamsters, of the Bio 14.6 and Bio 53.58 strains. Those hamsters were used as models for hypertrophic and dilated cardiomyopathy, respectively. The heart to body weight ratio in the Bio 14.6 strains was higher (P<0.05) than that in the age-matched F1b strains. In the Bio 53.58 strains, the heart to body weight ratio was higher at 8 and 42 weeks of age than that in the F1b strains. The collagen content increased from 22 weeks of age in both Bio hamsters compared with age-matched F1b hamsters (P<0.05). In both cardiomyopathic hamsters, the mRNA expressions for type I and type III collagen and fibronectin all increased with aging; however, the fibronectin expression in the Bio 14.6 strains increased more at 22 weeks of age than at 42 weeks of age. The left ventricular MMP-1, MMP-2 and MMP-9 activities in Bio 53.58 strains increased with aging. However, in the Bio 14.6 strains, although MMP-1 activities increased with aging, MMP-2 and MMP-9 activities decreased at 42 weeks of age in comparison to those at 22 weeks of age. Thus, the MMP activation differed between two cardiomyopathic models at the stage of heart failure, although the collagen synthesis was elevated in both models. In conclusion, it would seem that the relative balance between the synthesis and the removal of collagen may contribute to the changes in the left ventricular geometry in two different types of cardiomyopathy. PMID- 10471346 TI - Expression of T-type Ca(2+) channels in ventricular cells from hypertrophied rat hearts. AB - In this study we examined the existence of T-type Ca(2+) current in ventricular myocytes isolated from rats with pressure-overload hypertrophy. The whole-cell clamp technique was used to record Ca(2+) currents in enzymatically dissociated ventricular cells. T- and L-type Ca(2+) currents were separated by applying voltage steps to different test potentials from a holding potential of -80 mV and -50 mV. T-type Ca(2+) current was defined as the difference between the currents from the two holding potentials. Ventricular myocytes from sham-operated rats showed only L-type Ca(2+) current (maximal density -13.9+/-1.3 pA/pF n=17), whereas ventricular myocytes isolated from rats with aortic stenosis showed both L- and T-type Ca(2+) currents. The average values of T- and L-type Ca(2+) current density were -4.8+/-0.4 pA/pF and -12.4+/-0.9 pA/pF (n=32), respectively. T-type Ca(2+) current was distinguished from L-type Ca(2+) current by its voltage dependence, its kinetics and by its strong blockade by nickel 50 microM. In conclusion, we have demonstrated that hypertrophied ventricular rat cells express T-type Ca(2+) channels and this finding strongly supports a role for this channel in regulating growth processes in cardiac tissue. PMID- 10471347 TI - The expression of TGF-beta receptors in human atherosclerosis: evidence for acquired resistance to apoptosis due to receptor imbalance. AB - The degree of cellularity in vascular lesions is determined by the balance between the migration and proliferation of cells relative to their rate of egress and apoptosis. Transforming growth factor-beta(1) can act as a potent antiproliferative and apoptotic factor for proliferating vascular cells. Our laboratory has previously identified cells cultured from human vascular lesions that are resistant to the antiproliferative effect of TGF-beta(1) due to an acquired mutation in the Type II receptor for TGF-beta(1). In the present studies, the expression of the Type I and II receptors in coronary and carotid atherosclerotic lesions was analysed by immunostaining, RT-PCR, and in situ RT PCR. Levels of the Type I and Type II receptors varied widely within lesions, with the highest levels in the fibrous cap and at discrete foci within the lesion. Regions of smooth muscle-like cells (SMC) were commonly found that were Type I positive but Type II receptor negative. In 43 cell lines cultured from 126 human lesions, 84% of the lesion-derived cell (LDC) cultures exhibited functional resistance to the antiproliferative effect of TGF-beta(1). This resistance was conferred against TGF-beta(1), TGF-beta(2), and TGF- beta(3), but not interferon gamma or mimosine. While normal SMC exhibited a four-fold increase in the rate of apoptosis after TGF- beta(1) treatment, most LDC were resistant to apoptosis in response to TGF-beta(1). Resistant cells exhibited selective loss of Type II receptor expression, and retroviral transfection of Type II receptor cDNA partially corrected the functional deficit. Thus, resistance to apoptosis may lead to the slow proliferation of resistant cell subsets, thereby contributing to the progression of atherosclerotic and restenotic lesions. PMID- 10471348 TI - Multi-dose crystalloid cardioplegia preserves intracellular sodium homeostasis in myocardium. AB - The goal of this study was to assess the effect of multi-dose St Thomas' cardioplegia on intracellular sodium homeostasis in a rat heart model. A new magnetic resonance method was applied which enable us to detect intracellular Na changes without chemical shift reagents. Three groups of isolated rat hearts were subjected to 51 min of ischemia and 51 min of reperfusion at 37 degrees C: Group 1-three infusions of St Thomas' cardioplegia every 17 min for 2 min (n=7); Group 2-single-dose infusion of cardioplegia at the beginning of stop-flow ischemia (n=8); and Group 3-clamp ischemia (n=3) without cardioplegia administration. Performance of the heart was assessed by rate-pressure product relative to the pre-ischemic level (RPP). An NMR method was applied which continuously detects the Na(i) concentration in the heart, using the ability of bound sodium to exhibit triple-quantum transitions and the growth of the corresponding signal when sodium ions pass from extracellular to intracellular space. Clamp ischemia without cardioplegia and 50 min of reperfusion left the heart dysfunctional, with Na(i) growth from the pre-ischemic level of 13.9+/-1.2 mM to 34.9+/-1.3 mM and 73. 9+/-1.9 mM at the end of ischemia and reperfusion, respectively. During single-dose cardioplegia the corresponding values for Na(i) were 30.2+/-1 mM and 48.5+/-1.7 mM (RPP=29%). Multiple infusions of cardioplegic solution resulted in a remarkable preservation of the heart's intracellular Na concentration with a non-significant increase in Na(i) during ischemia and only 16.7+/-1 mM, (P=0.01), after subsequent reperfusion (RPP=85%). The time course of Na(i) changes in the rat heart model demonstrates a prominent potential of multi-dose St Thomas' cardioplegia in preserving intracellular sodium homeostasis at 37 degrees C. The growth of Na(i) concentration during ischemia, as an indicator of the viability of the myocytes, can have a prognostic value for the heart's performance during reperfusion. PMID- 10471349 TI - Trace amounts of albumin protect against ischemia and reperfusion injury in isolated rat hearts. AB - Albumin is used to provide colloid osmotic pressure in some resuscitation and organ preservation protocols. These solutions are expensive and carry the risks of using high concentrations of blood products. Used as a carrier of drugs and substrates, the concentration of albumin present in perfusates may be considerably lower in experimental ischemia. The present study examined if trace amounts of albumin (0.0004%) reduce injury from ischemia and reperfusion in isolated rat hearts. Hearts were perfused by the Langendorff technique (60 mmHg) with an intraventricular balloon. Zero-flow ischemia (20 min, 37 degrees C) was followed by reperfusion (35 min, 37 degrees C). Recovery of contractile function during reperfusion was significantly improved by the presence of fatty acid-free bovine serum albumin (BSA) (22 290+/-1280 mmHg/min, pressure-rate product) or rat serum albumin (RSA) (21 095+/-2836 mmHg/min) compared with Krebs-Henseleit buffer with no albumin (KHB) (9660+/-2324 mmHg/min). Release of lactate dehydrogenase activity, formation of tissue edema and accumulation of tissue malonyldialdehyde were significantly reduced in hearts receiving BSA or RSA compared with KHB alone. These parameters were not altered by the presence of albumin in non ischemic control hearts or in the pre-ischemic values of the hearts subjected to ischemia and reperfusion. Development of ischemic contracture with an extended period of ischemia (27 min) was not altered by the presence of BSA, suggesting that protection observed with albumin occurred during reperfusion, rather than during ischemia. Reperfusion following 45 min of ischemia with bovine serum albumin resulted in similar myocardial injury to hearts that were reperfused following 20 min of ischemia without bovine serum albumin. Thus, trace amounts of albumin provide significant reduction in myocardial injury from ischemia and reperfusion, probably via antioxidant mechanisms. PMID- 10471350 TI - Captopril treatment improves the sarcoplasmic reticular Ca(2+) transport in heart failure due to myocardial infarction. AB - Although captopril, an angiotensin-converting enzyme (ACE) inhibitor, has been shown to exert a beneficial effect on cardiac function in heart failure, its effect on the status of sarcoplasmic reticulum (SR) Ca(2+) transport in the failing heart has not been examined previously. In order to determine whether captopril has a protective action on cardiac function, as well as cardiac SR Ca(2+)-pump activity and gene expression, a rat model of heart failure due to myocardial infarction was employed in this study. Sham operated and infarcted rats were given captopril (2 g/l) in drinking water; this treatment was started at either 3 or 21 days and was carried out until 8 weeks after the surgery. The untreated animals with myocardial infarction showed increased heart weight and elevated left ventricular end diastolic pressure, reduced rates of pressure development and pressure fall, as well as depressed SR Ca(2+) uptake and Ca(2+) stimulated ATPase activities in comparison with the sham control group. These hemodynamic and biochemical changes in the failing hearts were prevented by treatment of the infarcted animals with captopril. Likewise, the observed reductions in the SR Ca(2+) pump and phospholamban protein contents, as well as in the mRNA levels for SR Ca(2+) pump ATPase and phospholamban, in the failing heart were attenuated by captopril treatment. These results suggest that heart failure is associated with a defect in the SR Ca(2+) handling and a depression in the gene expression of SR proteins; the beneficial effect of captopril in heart failure may be due to its ability to prevent remodeling of the cardiac SR membrane. PMID- 10471351 TI - Action potentials that mimic fibrillation activate sodium current. AB - Ventricular fibrillation (VF) has brief action potentials (50-70 ms) with short diastolic intervals (10-30 ms). Under these conditions ion channel activity may be grossly different to normal sinus rhythm (NSR). In particular, sodium channel activation may not contribute to the generation and propagation of action potentials during VF. This study determined if sodium channels can be activated when action potentials mimic VF. Isolated chick ventricular myocytes (n=7) were voltage-clamped to quantitate fast inward sodium current. The voltage clamp protocol simulated VF with a 10 pulse train at 10 Hz (100 ms cycle length (CL)) and depolarization interval (action potential duration) ranging from 90 to 20 ms. After each train a test pulse was delivered from holding (-80 mV) in 10-ms steps. The train preceded each step pulse. Peak sodium current for control and each VF protocol occurred at a membrane potential (V(m)) of -10 mV. Sodium current was evident during brief resting intervals as short as 20 ms, albeit 10-20% of baseline. Resting intervals less than 60 ms shifted the sodium conductance activation curve from Vm(0.5)-30 mV to -22 mV membrane potential. Similar findings occurred when resting potential was at -65 mV, although there was less sodium current with all tested protocols. There was significantly less inactivation of sodium current when the prepulse was shorter (100 v 1000 ms). There was approximately 20% greater sodium current when the test pulse followed a short v long depolarized (>-80 mV) prepulse. Although the longer depolarization pulses produce approximately 20% greater sodium current at membrane potentials more negative than -80 mV. Lastly the time for half recovery of sodium current from activation was significantly less when the inactivating prepulse was short v long (45.9+/-9 v 118+/-20 ms, P<0.05). In conclusion, sodium current is evident when the diastolic rest interval is as brief as 10-20 ms. Rest interval, length of membrane depolarization and membrane potential interact to affect sodium channel activation, inactivation and recovery from inactivation. These data demonstrate that the brief action potentials at more depolarized membrane potentials seen during VF allow for inward sodium current upon depolarization, less sodium channel inactivation, and a faster recovery from inactivation, thereby compensating for a short diastolic rest interval. Therefore, it is likely that the inward sodium channel contributes to wave front propagation during ventricular fibrillation. PMID- 10471352 TI - Bioenergetic, functional and morphological consequences of postinfarct cardiac remodeling in the rat. AB - Despite recent advances in the treatment, severe chronic heart failure (CHF) remains a syndrome associated with high mortality. Therefore, the search for new agents to improve both patient symptoms and survival, as well as the pursuit for detailed knowledge about pathophysiology of the failing heart, will continue to depend on relevant animal models. Large acute myocardial infarction (MI) initiates complex changes in the geometrical, structural, and biochemical architecture of both infarcted and non-infarcted regions of ventricular myocardium, which can profoundly affect left ventricular function and prognosis. In this paper we present a new model for non-invasive cardiac (31)P MRS in the rat. Volume-selective (31)P magnetic resonance spectroscopy and echocardiography were used for evaluation of myocardial energy metabolism, cardiac morphology and function in rats 3 days and 3 weeks after induction of large MI. The phosphocreatine:adenosine triphosphate (PCr:ATP) ratio was decreased in rats with MI comparing with controls both at 3 days (1.6+/-0.06 vs 2.7+/-0.04; mean+/ s.e.m. P<0.0001) and 3 weeks (1.6+/-0.07 v 2.7+/-0.02 P<0.0001) postinfarct. The results from the study demonstrate that postinfarct cardiac remodeling is a rapid process of changes not only in cardiac geometry, structure and function but also in myocardial energy metabolism after large transmural MI in the rat. PMID- 10471353 TI - Induction of cardiac nitric oxide synthase 2 in rats exposed to chronic hypoxia. AB - Induction of nitric oxide synthase (NOS2, also designated as iNOS) in the heart is known to occur in response to various stimuli. It is not known, however, whether in vivo hypoxia leads to cardiac NOS2 induction. We thus investigated the effects of normobaric hypoxia (10% O(2)for 8, 15 and 21 days) on NOS2 protein expression and enzyme activity in rat right ventricle (RV) and left ventricle (LV). Chronic hypoxia induced RV hypertrophy: the RV weight to body weight ratio was increased by 45% upon 15 days of exposure, with no change thereafter and no change in left ventricular (LV) weight. Treatment of hypoxic rats with l -NAME for 1 month decreased pulmonary artery pressure and RV hypertrophy compared to hypoxic non-treated rats. NOS2 activity detected by [(3)H]l -arginine to [(3)H]l citrulline conversion increased in RV during hypoxia, with a maximum at 15 days (+161% of control rats P<0.05), whereas it increased less (by 60%) in LV. In parallel, after 15 days of hypoxia there was a three-fold increase in NOS2 protein abundance detected by Western blotting using an isoform-specific antibody in the RVs (two-fold increase in the LV). Immunochemistry with the specific antibody demonstrated the expression in cardiomyocytes isolated from both ventricles of normoxic and hypoxic rats. Protein kinase C (PKC) content and activity was unchanged in LV of hypoxic rats, but increased in RV as compared with normoxic rats. These results clearly show that, in the heart, NOS2 is upregulated by hypoxia with an expression in cardiomyocytes of both ventricles. In addition, NOS2 is more inducible in the right hypertrophied ventricle than in the left non-hypertrophied hypoxic ventricle. PMID- 10471354 TI - Caspase inhibition reduces myocyte cell death induced by myocardial ischemia and reperfusion in vivo. AB - Myocardial ischemia and reperfusion lead to myocyte cell death, at least in part, by an apoptotic mechanism. Caspases are a conserved family of proteases that play an essential role in the execution of apoptosis; however, their potential contribution to ischemic myocardial cell death is largely unknown. To examine their role in this process, we subjected rabbits to 30 min of coronary artery occlusion followed by 3 h of reperfusion. Immunoblot analyses revealed that caspases-2, -3 and -7 were proteolytically activated during myocardial ischemia and reperfusion in vivo. In addition, the well-characterized caspase substrate poly(ADP-ribose) polymerase (PARP) was selectively cleaved into its signature apoptotic fragment in ischemic/reperfused myocardium. Systemic administration of the broad-spectrum caspase inhibitor acetyl-Tyr-Val-Ala-Asp chloromethylketone (YVAD-cmk, 4.8 mg/kg) partially blocked caspase activation and dramatically reduced the percentage of terminal dUTP deoyxynucleotidyl-transferase nick end labeling (TUNEL)-positive myocyte nuclei in the infarct region (3.9+/-0.8%v 13.0+/-2.2% in control animals, P=0.012). Moreover, YVAD-cmk reduced myocardial infarct size by approximately 31% (31.1+/-3.3%v 45.3+/-4.9% in control animals, P=0.032). These results indicate that caspases are critical mediators of myocardial injury induced by ischemia and reperfusion in vivo, and they suggest that caspase inhibition may be therapeutically beneficial in myocardial infarction. PMID- 10471355 TI - Developmental regulation of the translational repressor NAT1 during cardiac development. AB - The process of translation initiation has been postulated to play an important role in the regulation of cellular growth and proliferation. Here, we report the identification and differential expression of a fundamental translational repressor NAT1, during early postnatal cardiac development. Differential display analysis of RNA obtained from 3-day and 4-week-old rat hearts resulted in the cloning and identification of a 396 bp cDNA fragment (DRCF-6) which corresponded to the 3' terminal portion of NAT1. Northern blot analysis revealed that the mRNA expression of NAT1 was markedly elevated during the first 2 weeks of postnatal life, with an apparent peak level of expression occurring at 1 week. NAT1 mRNA levels then steadily decreased to 4 weeks of age. The NAT1 transcript has previously been shown to be extensively edited by the enzyme APOBEC-1, which deaminates specific cytidine bases to uridine; cytidine deamination at a glutamine codon (CAA) results in the formation of a stop codon (UAA) and consequently, premature termination of translation. Accordingly, Western blot analysis detected the presence of several smaller proteins in addition to the full length NAT1 protein (97 kDa), each exhibiting a distinct pattern of expression during cardiac development. APOBEC-1 editing of NAT1 during cardiac development was further supported by primer extension analysis of cytidine 1699, which was found to be predominantly edited to uridine. Immunohistochemical staining showed that NAT1 is expressed predominantly in atrial and ventricular myocytes, although staining was also detected in vascular smooth muscle cells and in the endocardium. These results suggest that NAT1 may play a role in the postnatal development of the heart and demonstrate that APOBEC-1 editing may possibly be a novel mechanism by which translation is regulated during cardiac development. PMID- 10471356 TI - The effect of isoproterenol on phospholamban-deficient mouse hearts with altered thyroid conditions. AB - The aim of the present study was to determine the effects of beta -adrenergic stimulation in wild-type and phospholamban-deficient mouse hearts with altered thyroid conditions. Hypothyroidism was associated with significant decreases in heart/body weight ratio in wild-type and phospholamban-deficient mice, whereas hyperthyroidism was associated with significant increases in heart/body weight ratio in both groups. Hypothyroid hearts of wild-type and phospholamban-deficient mice exhibited similar increases in beta -myosin heavy chain protein levels and decreases in alpha -myosin heavy chain protein levels. In hyperthyroidism, there were increases in the alpha -myosin heavy chain protein levels and these were similar in wild-type and phospholamban-deficient hearts. There were no detectable levels of beta -myosin heavy chain protein in the hyperthyroid hearts. The relative tissue level of phospholamban in wild-type hearts was increased (133%, P<0.01) in hypothyroidism, and decreased (69%, P<0.01) in hyperthyroidism, when compared to euthyroid controls (100%). Similar increases and decreases in SR Ca(2+)-ATPase protein levels were observed between phospholamban-deficient and wild-type hearts in hyperthyroidism and hypothyroidism, respectively. The basal contractile state of wild-type and phospholamban-deficient hearts was significantly depressed in hypothyroidism. On the other hand, the basal contractile state of wild-type and phospholamban-deficient hearts was significantly increased in hyperthyroidism. During beta -agonist stimulation of wild-type hearts, the responses in the rates of contraction and relaxation were highest in the hypothyroid group, followed by the euthyroid, and lastly by the hyperthyroid groups. There was a close linear correlation between the magnitude of the contractile parameter responses and the phospholamban/SERCA2 ratios in these hearts. However, the phospholamban-deficient hypothyroid, euthyroid, and hyperthyroid hearts did not exhibit any responses to isoproterenol, indicating that the alterations in the thyroid states of these hearts do not influence the effects of isoproterenol on cardiac function. These findings suggest that phospholamban is an important regulator of the heart's responses to beta adrenergic stimulation under various thyroid states. PMID- 10471357 TI - Role of slowed Ca(2+) transient decline in slowed relaxation during myocardial ischemia. AB - The goal of this study was to test the hypothesis that during myocardial ischemia, slowing of the Ca(2+) transient decline causes slowed relaxation. Our approach was to monitor pressure and Ca(2+) transients in isovolumic rat hearts during control and low flow ischemia conditions. In addition, we experimentally slowed the decline of the Ca(2+) transient using cyclopiazonic acid (CPA) to inhibit the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA, the most important pump for rapidly transporting Ca(2+) out of the cytosol). Using 9 microm CPA during normoxia, we were able to reproduce the slowed Ca(2+) transient decline and slowed relaxation found during low flow ischemia. The time constants of cytosolic [Ca(2+)] decline and pressure decline (tau(Ca) and tau(P) respectively) with CPA (78+/-5 ms and 64+/-3 ms) were similar to those found with ischemia (89+/-12 ms and 72+/-10 ms, mean+/-SEM, n=7) and were considerably greater than for controls (41+/-3 and 25+/-2 ms, mean+/-SEM, n=14, P<0.01). Furthermore, the relationship of tau(P) v tau(Ca) with CPA was similar to that found with ischemia. These findings are consistent with the hypothesis that the slowed Ca(2+) transient decline with both CPA and ischemia causes slowed relaxation. Consistent with this conclusion, a simple mathematical model to relate cytosolic [Ca(2+)] and pressure also suggests that slowed pressure relaxation can be explained by slowing of the Ca(2+) transient decline. This study suggests that impaired Ca(2+) uptake is a major injury causing slowed relaxation during ischemia. PMID- 10471358 TI - Chromosomal, in silico and in vitro expression analysis of cardiovascular-based genes encoding zinc finger proteins. AB - Three hundred and sixty expressed sequence tags (ESTs) from human heart cDNA libraries corresponding to one hundred and twenty six unique zinc finger proteins (ZFPs) were annotated and classified into seven types of ZFPs as reported previously. Among these 126 cvbZFPs (cardiovascular-based ZFPs), the C(2)H(2) type and the C(2)C(2)-type are the two major ZFP types which account for more than 80% of ZFP genes present in the cardiovascular system. The expression patterns of 11 randomly selected ZFP genes (at least one for each type) in normal fetal, adult and hypertrophic adult hearts, respectively, were determined using reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. The results suggest that ZFPs may be involved in the processes of either developmental control (downregulated or upregulated expression) or basic cellular functional regulation (constant expression). Interestingly, PAF-1 (peroxisome assembly factor-1), a C(3)HC(4)-type ZFP (RING domain-containing ZFP) showing a downregulated expression pattern in normal tissues was found to be upregulated in hypertrophic adult heart, suggesting a possible role for this fetal gene in the pathogenesis of cardiac hypertrophy. In silico Northern analysis of 15 tissues showed that over 90% of cvbZFPs demonstrate widespread tissue distribution, suggesting the vast majority of ZFPs are functionally shared among tissues. The potential importance of transcriptional repressors in cardiovascular development and disease, such as HFHZ, was supported by the observation that one-third (39 of 126) of cvbZFPs possess this function. Of these, 26 are C(2)H(2)-type and the remaining 13 included 8 C(2)C(2)-type, 1 C(3)HC(4)-type, 1 C(2)HC(4)C(HD)-type, 2 C(3)H-type and 1 combination type. Of particular interest was the observation that ZFPs which contain a KRAB domain are the major subtype present (51. 3% of the total repressors in cvbZFPs). Chromosomal distribution analysis showed that mapping loci of cvbZFP genes are concentrated on chromosomes 1, 3, 6, 8, 10, 11, 12, 19 and X. In particular, chromosome 19 appears to be enriched in ZFP genes with C(2)H(2)-type as the predominant type present. Overall, this report provides a fundamental initial step toward understanding the potential role of ZFPs in regulating cadiac development and disease. PMID- 10471359 TI - Galpha(i2), Galpha(i3)and Galpha(o) are all required for normal muscarinic inhibition of the cardiac calcium channels in nodal/atrial-like cultured cardiocytes. AB - The cardiac L-type calcium current (I(Ca,L)) is an important regulator of myocardial contractility. It is activated by sympathetic stimulation and inhibited by parasympathetic activity via muscarinic acetylcholine receptors. Muscarinic inhibition of I(Ca,L) occurs via activation of pertussis toxin (PTX) sensitive heterotrimeric G-proteins. Although recent studies have shown that expression of G(oalpha) is important for this effect in adult mouse ventricular cells, two other PTX-sensitive G-proteins (G(i2) and G(i3)) are also expressed in cardiocytes and are activated. Their role in the regulation of I(Ca,L) has not been examined. In addition, it is not known whether nodal/atrial cardiac cells use the same G-proteins. We show that gene inactivation of each of the three PTX sensitive Galpha-proteins (alpha(i2), alpha(i3), and alpha(o)) affects muscarinic inhibition of cardiac I(Ca,L) in embryonic stem (ES) cell-derived cardiocytes. Inactivation of either alpha(i2) or alpha(i3) markedly slows the time course of muscarinic inhibition of I(Ca,L), and in cells where both alpha(i2) and alpha(i3) are inactivated the effects are not additive. We also establish an essential role for alpha(o)in this atrial/nodal-like cardiocyte system and show that alpha(o)acts proximal to NO generation. NO generation plays a critical role in I(Ca,L) regulation since the nitric oxide synthase (NOS) antagonist, l -NMMA, blocked the inhibition of I(Ca,L) in WT and in alpha(i2)/alpha(i3)-null cells. In WT cells, the NO generating agent SIN-1 inhibited I(Ca,L) and the addition of carbachol resulted in faster inhibition, suggesting that pathways in addition to NO are also activated. This study shows that alpha(i2) and alpha(i3) play a critical role in the normal inhibition of cardiocyte I(Ca,L). Thus, all muscarinic receptor activated G-proteins (G(i2), G(i3) and G(o)) are necessary for normal inhibition and act through both NO and non-NO signaling pathways. PMID- 10471360 TI - Conflict Aversion: Preference for Ambiguity vs Conflict in Sources and Evidence. AB - This research investigates preferences and judgments under ambiguous vs conflicting information. Three studies provided evidence for two major hypotheses: (1) Conflicting messages from two equally believable sources are dispreferred in general to two informatively equivalent, ambiguous, but agreeing messages from the same sources (i.e., conflict aversion); and (2) conflicting sources are perceived as less credible than ambiguous sources. Studies 2 and 3 yielded evidence for two framing effects. First, when the outcome was negative, subjects' preferences were nearly evenly split between conflict and ambiguity, whereas a positive outcome produced marked conflict aversion. Second, a high probability of a negative outcome or a low probability of a positive one induced conflict preference. However, no framing effects were found for source credibility judgments. Study 3 also investigated whether subject identification with a source might affect preferences or credibility judgments, but found no evi dence for such an effect. The findings suggest cognitive and moti vational explanations for conflict aversion as distinct from ambi guity aversion. The cognitive heuristic is that conflict raises suspicions about whether the sources are trustworthy or credi ble. The motivational explanation stems from that idea that if sources disagree, then the judge not only becomes uncertain but also must disagree with at least one of the sources, whereas if the sources agree then the judge may agree with them and only has to bear the uncertainty. Copyright 1999 Academic Press. PMID- 10471361 TI - The Influence of Physical Attractiveness and Gender on Ultimatum Game Decisions. AB - Physical appearance influences behavior in a number of environments, yet surprisingly little is known about the influence of physical attractiveness on the bargaining process. We conducted an ultimatum game experiment to investigate the influence of physical attractiveness and gender on ultimatum game decisions. Results from this study revealed no significant differences in the offers or demands attractive and unattractive people made. However, attractive people and men were treated differently by others. Consistent with the notion of a "beauty premium," attractive people were offered more, but more was demanded of them. Men were also offered more, and less was demanded of them. We discuss implications of these results with respect to bargaining and the labor market. Copyright 1999 Academic Press. PMID- 10471362 TI - Overconfidence: It Depends on How, What, and Whom You Ask. AB - Many studies have reported that the confidence people have in their judgments exceeds their accuracy and that overconfidence increases with the difficulty of the task. However, some common analyses confound systematic psychological effects with statistical effects that are inevitable if judgments are imperfect. We present three experiments using new methods to separate systematic effects from the statistically inevitable. We still find systematic differences between confidence and accuracy, including an overall bias toward overconfidence. However, these effects vary greatly with the type of judgment. There is little general overconfidence with two-choice questions and pronounced overconfidence with subjective confidence intervals. Over- and underconfidence also vary systematically with the domain of questions asked, but not as a function of difficulty. We also find stable individual differences. Determining why some people, some domains, and some types of judgments are more prone to overconfidence will be important to understanding how confidence judgments are made. Copyright 1999 Academic Press. PMID- 10471363 TI - Culture and Negotiator Cognition: Judgment Accuracy and Negotiation Processes in Individualistic and Collectivistic Cultures. AB - In this paper, we argue that judgment biases in negotiation are perpetuated by underlying cultural values and ideals, and therefore, certain judgment biases will be more prevalent in certain cultural contexts. Based on theory in cultural psychology (Markus & Kitayama, 1991; Triandis, 1989), we considered the notion that fixed pie error, a judgment bias in which negotiators fail to accurately understand their counterparts' interests (Pruitt & Lewis, 1975; Thompson & Hastie, 1990), would be more prevalent at the end of negotiations in the United States, an individualistic culture, than Greece, a collectivistic culture. The results of a 2-week computer-mediated intercultural negotiation experiment, which took place between American students in Illinois and Greek students in Athens, supported this view. Theoretical implications of culture and cognition in negotiation are also discussed. Copyright 1999 Academic Press. PMID- 10471364 TI - A new role of ran GTPase. AB - Ran is a G protein similar to Ras, but it has no membrane binding site. RanGEF, RCC1, is on chromatin and RanGAP, RanGAP1/Rna1p is in cytoplasm. Ran, thus, shuttles between the nucleus and the cytoplasm to complete its GTPase cycle, carrying out nucleocytoplasmic transport of macromolecules. A majority of Ran binding proteins, thus far found, are required for this process. A recently found novel Ran-binding protein, RanBPM, however, is localized in the centrosome. Subsequently, four groups reported that RanGTP, but not RanGDP, can induce microtubule self-organization in Xenopus egg extracts where no nuclear membrane is present. Thus, Ran is suggested to have a new role beyond the nucleocytoplasmic transport of macromolecules. In both microtubule assembly and nucleocytoplasmic transport, chromosomal localization of RCC1 is important to carry out the functions of RanGTPase. In this regard, a future intriguing question is how RCC1 interacts with chromatin DNA. PMID- 10471365 TI - Identification of four new members of the rat prolactin/growth hormone gene family. AB - The rodent prolactin (PRL)/growth hormone (GH) gene family currently consists of at least 14 distinct genes that are expressed mainly in pituitary, uterus, and/or placenta. We report here the identification of novel four members from rat with significant homology to PRL. The encoding proteins are not homologs of other known members of this hormone family. The four new cDNAs were assigned to PRL family based on sequence homology and were referred to as PRL-like protein-I (PLP I), PLP-J, PLP-K, and PLP-L, following the current naming order of rodent PLP family, where PLP-H is the most recent gene. They encode amino acids with 211-228 amino acids, and 34-38% identity with PRL. All have one or two N-linked glycosylation sites. Among the examined rat tissues by Northern blot analysis, only PLP-I was expressed in testis. Our results indicate that the rodent PRL/GH gene family is large with at least 18 distinct genes. PMID- 10471366 TI - 15-Deoxy-Delta(12,14)-prostaglandin J(2), a ligand for peroxisome proliferator activated receptor-gamma, induces apoptosis in JEG3 choriocarcinoma cells. AB - Apoptosis has been described in placental (trophoblast) tissues during both normal and abnormal pregnancies. We have studied the effects of the cyclopentenone prostaglandins (PGs) on trophoblast cell death using JEG3 choriocarcinoma cells. PGJ(2), Delta(12)PGJ(2), and 15-deoxy-Delta(12,14)-PGJ(2) (15dPGJ(2)) (10 microM) significantly reduced mitochondrial activity (MTT assay) over 16 h by 17.4 +/- 4.7%, 28 +/- 9.3%, and 62.5 +/- 2.8%, respectively (mean +/ sem), while PGA(2) and PGD(2) had no effect. The synthetic PPAR-gamma ligand ciglitizone (12.5 microM) had a potency similar to 15dPGJ(2) (69 +/- 3% reduction). Morphological examination of cultures treated with PGJ(2) and its derivatives revealed the presence of numerous cells with dense, pyknotic nuclei, a hallmark of apoptosis. FACS analysis revealed an abundance (approximately 40%) of apoptotic cells after 16-h treatment with 15dPGJ(2) (10 microM). The caspase inhibitor ZVAD-fmk (5 microM) significantly diminished the apoptotic effects of Delta(12)PGJ(2) and 15dPGJ(2). JEG3 cells expressed PPAR-gamma mRNA by Northern analysis. These novel findings imply a role for PPAR-gamma ligands in various processes associated with pregnancy and parturition. PMID- 10471367 TI - Protein-induced fusion of phospholipid vesicles of heterogeneous sizes. AB - We have investigated the fusion of phospholipid vesicles induced by lysozyme and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Vesicles were composed of dimyristoylphosphatidylcholine/dioleoylphosphatidylethanolamine/ cholesterol (DMPC:DOPE:Chol, 2:1:1). Small unilamellar vesicles (SUV, diameter ca. 30 nm) obtained by extensive sonication or large unilamellar vesicles (LUV, diameters ranged from 100 to 400 nm) obtained by extrusion methods were used. Fusion of LUV induced by lysozyme and GAPDH was drastically decreased when the diameter of the vesicles increased over a value of 100 nm. Lysozyme effect was stopped at the aggregation step while GAPDH effect was stopped at the fusion (lipid mixing) step. Fusion of heterogeneous vesicle populations (SUV with LUV) was observed only with GAPDH and this happened only when the lipids were in the liquid crystalline state. PMID- 10471368 TI - Compounds that induce isoforms of glutathione S-transferase with properties of a critical enzyme in defense against oxidative stress. AB - Compounds that upregulate enzymes that play critical roles in protection against free radical damage might be useful in treating diseases in which free radicals are pathological. To identify critical enzymes and their upregulators, compounds that were not free radical scavengers were screened for the ability to increase the IC(50) of the human neuronal cell line IMR-32 for hydrogen peroxide. Subsequently, enzymes upregulated by compounds that increased the IC(50) were identified. All of the compounds identified that increased the IC(50) also increased the specific activity of glutathione S-transferase (GST). In addition, compound-caused increases in the specific activity of GST correlated with compound-caused increases in the IC(50), the expected behaviour if GST was a critical enzyme. The GST isoform composition changed on upregulation, suggesting the upregulation of isoforms with anti-free radical activities. Structural features of compounds concurrently increasing the IC(50) and upregulating GST were identified. PMID- 10471369 TI - CD39 as a caveolar-associated ectonucleotidase. AB - CD39 is a human lymphoid cell activation antigen, (also referred to E-ATPDase or apyrase) that hydrolyzes extracellular ATP and ADP. Although it has been widely studied, its physiological role, however, still remains unclear. This ectonucleotidase generally is said to be evenly distributed in the membrane of the cells. However, we observed that in cell types which possess caveolae, specialised membrane invaginations involved in signalling, CD39 is preferentially targeted to these membrane microdomains. Since all molecules involved in signalling (eNOS, G-proteins, receptors) which are targeted to the caveolae undergo posttranslational modifications (e.g., palmitoylation) we hypothesize the same to be the case for CD39. Furthermore, its presence in the caveolae supports its participation in signalling events. PMID- 10471370 TI - Diethanolamine inhibits choline uptake and phosphatidylcholine synthesis in Chinese hamster ovary cells. AB - Diethanolamine (DEA), an alkanolamine used widely in industry, is hepatocarcinogenic in mice. The goal of this work was to determine whether DEA altered choline homeostasis in cultured cells, so as to ascertain whether the liver tumor response may be related to choline deficiency. CHO cells were cultured in Ham's F-12 medium containing DEA (0-1000 microgram/ml) and [(33)P] phosphorus was used to label phospholipid pools. After 48 hours incubation, lipids were extracted and [(33)P]-labeled phospholipids were quantified by autoradiography after thin layer chromatographic separation. In control cells, phosphatidylcholine (PC) accounted for 51 +/- 0.7% of the total lipid (33)P incorporation. DEA had no effect on cell number or total phospholipid biosynthesis, but it significantly decreased the incorporation of (33)P into PC at concentrations >/=50 microgram/ml. DEA (>/=20 microgram/ml) also inhibited the uptake of [(3)H]-choline into CHO cells, with 95% inhibition observed at 250 microgram/ml. To determine whether supplemental choline prevented PC synthesis inhibition by DEA, CHO cells were cultured with or without excess choline (30 mM) and DEA (500 microgram/ml). DEA reduced PC synthesis to 27 +/- 3% of total phospholipids, but had no effect on PC synthesis in choline-supplemented cells. When [(14)C]-DEA was incubated with CHO cells, it was also incorporated into the phospholipid fraction. Collectively, these results indicate that DEA reversibly inhibits PC synthesis by blocking choline uptake and competing for utilization in the CDP-choline pathway in CHO cells. PMID- 10471371 TI - Hydrogen peroxide in human urine: implications for antioxidant defense and redox regulation. AB - The presence of hydrogen peroxide, at levels sometimes exceeding 100 microM, in human urine samples was established by three different assay methods: 2 oxoglutarate decarboxylation and the ferrous oxidation-xylenol orange (FOX) assay and an oxygen electrode. Detected levels of H(2)O(2) were decreased by addition of superoxide dismutase. We conclude that urine contains autooxidizable molecules that, upon exposure to 21% O(2), undergo rapid superoxide-dependent autooxidation reactions to generate H(2)O(2). The exposure of human tissues to hydrogen peroxide may be greater than is commonly supposed, which has implications in relation to the proposed role of this species in cell signaling. PMID- 10471372 TI - Migration of exogenous immature hematopoietic cells into adult mouse brain parenchyma under GFP-expressing bone marrow chimera. AB - Bone marrow transplantation with GFP-expressing cells from GFP-transgenic mice resulted in migration of GFP-positive cells into peripheral tissues and brain parenchyma. Most of these cells were observed as colony-like clusters. GFP positive clusters in the brain were stained by antibody for ER-MP12, but those in the peripheral tissues were not. Since ER-MP12 antigen has been reported as a marker for murine early-stage myeloid precursor, this might suggest that some parts of phagocytic cells in the brain parenchyma such as microglia are derived from undifferentiated pluripotent hematopoietic cells. PMID- 10471373 TI - Proteolytic fragments of anti-HIV and anti-tumor proteins MAP30 and GAP31 are biologically active. AB - We analyzed the structural and functional organization of anti-HIV and anti-tumor proteins MAP30 and GAP31 by limited proteolysis with endopeptidases Lys-C and Glu C (V8). MAP30 and GAP31 are resistant to proteolytic digestion under conditions of as much as 5% (w/w) proteases. In the presence of 10% (w/w) protease, the central regions of the proteins are still resistant to proteolysis, whereas the N and C-termini are accessible. Peptide fragments were purified by FPLC on Superdex 75 columns, characterized by gel electrophoresis, identified by amino acid sequencing, and analyzed for anti-HIV, anti-tumor, and other biochemical activities. We report here that limited proteolysis yields biologically active fragments of both MAP30 and GAP31. These fragments are active against HIV-1 and tumor cells with EC(50)s in the sub-nanomolar ranges, 0.2-0.4 nM. At the dose levels used in the assays, little cytotoxicity to normal cells was observed. In addition, these fragments remain fully active in HIV-integrase inhibition and HIV LTR topological inactivation, but not ribosome inactivation. These results demonstrate that the antiviral and anti-tumor activities of MAP30 and GAP31 are independent of ribosome inactivation activity. In addition, we demonstrate that portions of the N- and C-termini are not essential for antiviral and anti-tumor activities, but do appear to be required for ribosome inactivation. These results may provide novel strategies for rational design and targeted development of mimetic antiviral and anti-tumor therapeutics. PMID- 10471374 TI - A sensitive new method for rapid detection of abnormal methylation patterns in global DNA and within CpG islands. AB - To assess alterations in DNA methylation density in both global DNA and within CpG islands, we have developed a simple method based on the use of methylation sensitive restriction endonucleases that leave a 5' guanine overhang after DNA cleavage, with subsequent single nucleotide extension with radiolabeled [(3)H]dCTP. The methylation-sensitive restriction enzymes HpaII and AciI have relatively frequent recognition sequences at CpG sites that occur randomly throughout the genome. BssHII is a methylation sensitive enzyme that similarly leaves a guanine overhang, but the recognition sequence is nonrandom and occurs predominantly at unmethylated CpG sites within CpG islands. The selective use of these enzymes can be used to screen for alterations in genome-wide methylation and CpG island methylation status, respectively. The extent of [(3)H]dCTP incorporation opposite the exposed guanine after restriction enzyme treatment is directly proportional to the number of unmethylated (cleaved) CpG sites. The "cytosine-extension assay" has several advantages over existing methods because (a) radiolabel incorporation is independent of the integrity of the DNA, (b) methylation detection does not require PCR amplification or DNA methylase reactions, and (c) it is applicable to ng quantities of DNA. Using DNA extracted from normal human liver and from human hepatocellular carcinoma, the applicability of the assay is demonstrated by the detection of an increase in genome-wide hypomethylation and CpG island hypermethylation in the tumor DNA. PMID- 10471375 TI - Sequencing the gene encoding desulfovibrio desulfuricans ATCC 27774 nine-heme cytochrome c. AB - Contradicting early suggestions, the sequencing of the gene encoding the Desulfovibrio desulfuricans (ATCC 27774) nine-heme cytochrome c proves that this cytochrome is not the product of the degradation of the 16-heme containing cytochrome c [Coelho et al. (1996) Acta Cryst. D52, 1202-1208]. However, preliminary data indicate that the cytochrome gene is part of an operon similar to the DvH hmc operon, which contains the gene coding for the 16-heme cytochrome c [Rossi et al. (1993) J. Bacteriol. 175, 4699-4711]. Also, the amino acid sequence deduced from the DNA sequence shows four residues in the C-terminal not predicted in the amino acid sequence obtained by X-ray methods [Matias et al. (1999) Structure 7, 119-130]. PMID- 10471376 TI - Functional analysis of four CYP21 mutations from spanish patients with congenital adrenal hyperplasia. AB - Deleterious mutations in the CYP21 (steroid 21-hydroxylase) gene cause congenital adrenal hyperplasia (CAH). These mutations usually result from recombinations between CYP21 and an adjacent pseudogene, CYP21P, including deletions and transfers of deleterious mutations from CYP21P to CYP21 (gene conversions). Additional rare mutations that are not gene conversions account for 5-10% of 21 hydroxylase deficiency alleles. Recently, four novel CYP21 point mutations leading to amino acid changes were identified in a population of 57 Spanish families with CAH. A nonsense mutation, K74X, was also identified. The enzymatic activities of 21-hydroxylase mutants G90V, G178A, G291C, and R354H were examined in transiently transfected CHOP cells using progesterone and 17alpha hydroxyprogesterone as substrates. The G90V, G291C, and R354H mutations effectively eliminated 21-hydroxylase activity. However, the G178A mutant retained significant activity when 17alpha-hydroxyprogesterone was the substrate. These results correlate well with the identification of G90V, G291C, and R354H in patients with severe "salt-wasting" disease and G178A in a patient with the milder simple virilizing form. PMID- 10471377 TI - Targeting of green fluorescent protein expression to the cell surface. AB - We have previously reported on GPI-anchored fusion proteins that bind radioactive isotopes. We targeted their expression to the cell surface to obtain a marker protein detectable by nuclear and optical imaging (1, 2). Here we suggest a novel approach for targeting a model protein (GFP) to the exoplasmic surface of the plasma membrane. An expression vector (pcPEP-GFP) was constructed containing GFP cDNA fused with the fragment encoding the N-terminal cytoplasmic domain and signal peptide/membrane anchoring domain of the rabbit neutral endopeptidase (PEP GFP). Flow cytometry showed green fluorescence in 45% of cells transfected with GFP and in 34% of cells transfected with PEP-GFP (24 h after transfection). Fluorescence microscopy of fixed cells stained with rhodaminated anti-GFP antibodies showed positive reaction only in the case of PEP-GFP-transfected cells indicating cell-surface expression. The PEP-GFP fusion protein was identified as a component of the light microsomal and Golgi fractions by immunoblotting. PMID- 10471378 TI - 6-Methyl-3'-bromoflavone, a high-affinity ligand for the benzodiazepine binding site of the GABA(A) receptor with some antagonistic properties. AB - 6-Methyl-3'-bromoflavone inhibited [(3)H]flunitrazepam binding to the benzodiazepine binding site of the GABA(A) receptor (BDZ-bs) with Ki values between 10 and 50 nM in different brain regions. The GABA ratio of 1.03 for [(3)H]flunitrazepam binding to cerebral cortex, 0.76 for cerebellum, 0.7 for hippocampus, 0.7 for striatum, and 0.8 for spinal cord indicated an antagonistic or weak inverse agonistic profile of 6-methyl-3'-bromoflavone on BDZ-bs. Unlike classical benzodiazepines, it had no anticonvulsant, anxiolytic, myorelaxant, sedative, amnestic or motor incoordination effects. However, it antagonized the muscle relaxant, the sedative effect, and the changes in locomotor activity induced by diazepam. Taken together, these findings suggest that 6-methyl-3' bromoflavone has an antagonistic profile on the BDZ-bs. PMID- 10471379 TI - Singlet oxygen ((1)Delta(g)O(2)) as the principal oxidant in myeloperoxidase mediated bacterial killing in neutrophil phagosome. AB - Intraphagosomal viability of wild type E. coli and lycopene (a powerful (1)O(2) quencher)-producing transformant E. coli was investigated using human polymorphonuclear leukocytes as the cells for phagocytosis of opsonized viable bacteria. While the viability of both wild type and the transformant E. coli decreased very rapidly in the phagosome, but the viability of the lycopene transformant in phagosomes was about 1.7 times higher than that of wild type E. coli after 5 min of incubation. The results were very similar to the results obtained when E. coli strains were exposed to (1)O(2) generated in myeloperoxidase-H(2)O(2)-Br(-) system (a pure (1)O(2) generating system) at pH 4.5. The reason for HOCl, which may be generated in the myeloperoxidase-H(2)O(2) Cl(-) system under physiological conditions but does not become involved in bactericidal action, could be explained by the near neutral pH in phagosomes at which bacterial killing by chlorination is extensively attenuated. This is the first report which proved (1)O(2)-mediated bacterial killing in neutrophil bacterial phagosomal system. PMID- 10471380 TI - Novel inhibitors of advanced glycation endproducts. AB - Enhanced formation and accumulation of advanced glycation endproducts (AGE's) have been proposed to play a major role in the pathogenesis of diabetic complications, aging, atherosclerosis, and Alzheimer disease leading to progressive and irreversible intermolecular protein crosslinkings. This process is accelerated in diabetes and has been postulated to contribute to the development of a range of diabetic complications including nephropathy, retinopathy and neuropathy. Several potential drug candidates as AGE inhibitors have been reported recently. Aminoguanidine is the first drug extensively studied both in vitro and in vivo. We have developed a new class of compounds as potent inhibitors of glycation and AGE formation. The novel inhibitors reported here are aryl (and heterocyclic) ureido, and aryl (and heterocyclic) carboxamido phenoxy isobutyric acids and related molecules, which were found by in vitro assay methods to be potent inhibitors of multiple stage of glycation and AGE formation. PMID- 10471381 TI - Collateral resistance of a dideoxycytidine-resistant cell line to 5-fluoro-2' deoxyuridine. AB - Exposure of a human lymphocytic cell line, H9 cells, to 0.5 microM and 5.0 microM dideoxycytidine (ddC) resulted in isolation of ddC-resistant H9-ddC0.5w and H9 ddC5.0w cell lines. In addition, these cell lines were also resistant to azidothymidine and had reduced deoxycytidine kinase and thymidine kinase activities. We now show that these cell lines are 4-fold and 2000-fold collaterally resistant to 5-fluoro-2'-deoxyuridine (FdUR), respectively, but not to 5-fluorouracil (FU). Biochemical evaluations show that, compared to the parental cells, the FdUR phosphorylation was reduced to 36.3% and 9.2% and the FdUMP levels were decreased to 48.1% and 1.2% in these cell lines. Taken together, the data suggest that ddC, an antiviral agent, is capable of inducing resistance to FdUR-a drug that is not its analog and which has a different metabolism, target site, and mechanism of action. PMID- 10471382 TI - The hydroxyl free radical reactions of ascorbyl palmitate as measured in various in vitro models. AB - The OH(*) free radical scavenging properties of ascorbyl palmitate (AP), water solubilized in the presence of a surfactant (Brij 35), were tested in various systems: (1) The inhibition of polymerization of bovine serum albumin by OH(*) free radicals generated by the Fenton reaction indicated AP exerts a considerable protective effect against polymerization by scavenging the OH(*) free radicals. (2) ESR spin trapping comparisons of DMPO with AP were conducted. Using the Fenton reaction as a source of OH(*) free radicals, AP was 1 order of magnitude faster in scavenging these radicals than DMPO. (3) Oxidative modification of BSA by (60)Co-gamma irradiation of 80 krad, results in a strong increase in protein carbonyl content. AP inhibits carbonyl formation very efficiently, indicating that AP may be utilized as a biological OH(*) free radical scavenger in human therapy. PMID- 10471383 TI - Cytoskeletal reorganization leads to induction of the urokinase-type plasminogen activator gene by activating FAK and Src and subsequently the Ras/Erk signaling pathway. AB - Previously, we showed that cytoskeletal reorganization (CSR) induced by colchicine or cyochalasins leads to activation of the urokinase-type plasminogen activator (uPA) gene in LLC-PK(1) cells via the Ras/Erk signaling pathway [Irigoyen et al. (1997) J. Biol. Chem. 272, 1904]. It remained to be seen how CSR activates Ras/Erk signaling. Changes in cell morphology triggered by extracellular signals are often mediated by integrin-associated proteins, such as focal adhesion kinase (FAK) and Src. We found that CSR induced the activation of FAK and Src and the association of FAK and Shc, a signaling molecule linking growth factor receptor tyrosine kinase and Grb2. Furthermore, expression of either FRNK, a kinase-minus FAK-like molecule acting as a dominant negative FAK, or a dominant negative Src suppressed CSR-induced uPA gene promoter activation. These results suggest that cells respond to a morphology change, using the cytoskeleton as a sensor, by activating FAK and Src and subsequently the Ras/Erk signaling pathway. PMID- 10471384 TI - DNA methylation analysis using bisulfite treatment and PCR-single-strand conformation polymorphism in colorectal cancer showing microsatellite instability. AB - The combination of bisulfite treatment and PCR-single-strand DNA conformation polymorphism (SSCP) analysis is proposed for quantitative methylation assay. We applied this procedure to the methylation analysis of the hMLH1 promoter region in colorectal cancer. An analysis of mixtures of known amounts of methylated and unmethylated DNA revealed a linear relation. Using a calibration curve, proportions of methylated DNA were calculated. The hMLH1 promoter region was highly methylated in about 80% of microsatellite instability (MSI) (+) colorectal cancers, but in none of the MSI(-) colorectal cancers. A significant correlation existed between hypermethylation of the hMLH1 promoter and MSI, as in previous reports. In conclusion, bisulfite-PCR-SSCP (BiPS) analysis could be applied to the rapid identification of methylation status in multiple samples, quantification of methylation differences, and detection of methylation heterogeneity in amplified DNA fragments. PMID- 10471385 TI - Identification of a differentially expressed RNA helicase by gene trapping. AB - A mouse line was generated that expressed a gene trap reporter construct, betageo, in a dynamic pattern during embryonic development. Differential expression was seen within the developing eyes, limbs, heart, neural tube, and skeleton. Two transcripts were cloned that contained endogenous sequences fused to the gene trap vector sequence. Analysis of the endogenous sequences revealed that the reporter integrated within a gene belonging to a small group of eukaryotic superfamily I helicases. Unexpectedly, the majority of transcripts produced from the trapped locus were not affected by the insertion of the reporter. Although the function of the trapped helicase gene is unknown, its complex transcription patterns and widespread spatial-temporal distribution suggest that the gene product plays a role in RNA metabolism in multiple tissues and organs within the developing embryo. PMID- 10471386 TI - Effects of exogenous nitric oxide and hyperoxia on lung fibroblast viability and DNA fragmentation. AB - Effective lung repair after acute injury requires elimination of proliferating mesenchymal and inflammatory cells without inducing an acute inflammatory response or disturbing concomitant repair of lung microvasculature. Previous studies have shown that endogenous NO regulates programmed cell death in fibroblasts and can modulate wound fibroblast synthetic function. We hypothesized that exposure of human lung fibroblasts to NO gas would decrease viability and induce apoptotic cell death. Primary cultures of normal human lung fibroblasts were exposed for 4 h to room air (RA), 80% oxygen, NO (at either 20 or 50 ppm) blended with RA, or NO blended with 80% O(2), then incubated for 24 to 72 h. Cell viability was determined by fluorescence viability/cytotoxicity assay and DNA fragmentation by TUNEL assay. Peroxynitrite formation was assessed using immunoblotting for S-nitrosotyrosine. NO plus O(2) induced significant cell death at 20 and 50 ppm NO when compared to either RA or O(2) alone at both 24 and 72 h (p < 0.05). Incubation with superoxide dismutase (SOD), catalase (CAT) or SOD + CAT significantly decreased cell death in fibroblasts treated with NO(20)/O(2) and NO(50)/O(2) compared with controls (p < 0.05). NO(20)/O(2) and NO(50)/O(2) exposure significantly increased TUNEL mean fluorescence intensity (MFI), consistent with increased DNA fragmentation, compared to RA at 24 and 72 h (p < 0.05). Antioxidants decreased MFI in cells exposed to NO(20)/O(2) (CAT and SOD + CAT) compared to controls at 24 h (p < 0.05). Western blot analysis for S nitrosotyrosine showed increased signal intensity in fibroblasts exposed to NO at 20 and 50 ppm plus O(2) compared to RA or O(2) alone. Incubation with SOD + CAT reduced signal intensity for peroxynitrite in cells exposed to NO(20)/O(2). We conclude that NO in hyperoxic conditions induces fibroblast cell death and DNA fragmentation, which could be partially mediated by peroxynitrite synthesis. PMID- 10471387 TI - Nuclear membrane sphingomyelin-cholesterol changes in rat liver after hepatectomy. AB - Sphingomyelin and cholesterol play an important role in stabilising the plasma membranes architecture and in many physiological process such as cell growth and differentiation. Degradation of sphingomyelin by exogenous sphingomyelinase induces a decrease of cholesterol due either to an increase of esterification or to a reduced biosynthesis. Variations of sphingomyelin due to the presence of a neutral-sphingomyelinase and of sphingomyelin-synthase have been recently shown in rat liver nuclear membranes. The aim of this research is to study the relation between sphingomyelin and cholesterol in the nuclear membranes following sphingomyelinase activation and during cell proliferation. The nuclear membranes, isolated from liver nuclei, were analysed for their content in protein, nucleic acids, and lipids (sphingomyelin and cholesterol) before and after sphingomyelinase activation and during hepatic regeneration. The activities of nuclear membrane SM-syntase and sphingomyelinase were also determined. The results confirmed that also in the nuclear membranes sphingomyelinase, especially exogenous, causes a strong decrease in cholesterol. The increase observed of sphingomyelin during the first 18 h after hepatectomy followed by a decrease at 24 h, due to the different activity of the enzymes, is accompanied by similar behaviour of cholesterol. This confirms the effect of neutral-sphingomyelinase on cholesterol, due to an increase of esterification process. Changes in cholesterol content modify the nuclear membranes fluidity and, as consequence, mRNA transport as previously shown. It can therefore be concluded that the neutral sphingomyelinase, present in the nuclei, may, across this mechanism, regulate the cell function. PMID- 10471388 TI - Transcriptional regulation of C/EBPdelta in G(0) growth-arrested mouse mammary epithelial cells. AB - Little is known about the control of cell cycle exit/G(0) entry, or the regulation of genes that are expressed during G(0). In this report we used primer extension analysis to demonstrate the high level of C/EBPdelta mRNA expression in G(0) growth-arrested HC11 mouse mammary epithelial cells and to identify the C/EBPdelta transcription start site. The C/EBPdelta gene transcription rate and promoter activity are both highly induced in G(0) growth-arrested HC11 cells. The C/EBPdelta gene promoter also exhibits G(0)-specific autoregulation. In contrast, the C/EBPdelta promoter activity decreases in G(0) growth-arrested NIH 3T3 cells. These data indicate that C/EBPdelta is among a relatively small number of genes actively transcribed during G(0) growth arrest. C/EBPdelta may regulate the expression of genes implicated in the initiation or maintenance of mammary epithelial cell G(0) growth arrest. PMID- 10471389 TI - Alkaline protease inhibitor: a novel class of antifungal proteins against phytopathogenic fungi. AB - A Streptomyces sp., which produces an alkaline protease inhibitor (API) exhibiting antifungal activity has been isolated from soil. The protein has been purified to homogeneity. The molecular characterization has revealed that it is a dimer (M(r) 28 kDa) with five disulphide linkages and has a pI of 3.8. API is a competitive type of inhibitor with a K(i) value of 2.5 x 10(-9) M. The inhibitor is stable over a pH range of 6 to 12 and a temperature range of 40 to 95 degrees C. API exhibits antifungal activity (in vitro) against phytopathogenic fungi such as Fusarium, Alternaria, and Rhizoctonia and also against Trichoderma, a saprophytic fungus. The antifungal activity of API appears to be associated with its ability to inhibit the fungal serine alkaline protease(s), which is indispensable for its growth. Retardation of the rate of fungal spore germination, as well as hyphal extention, was observed in the presence of API. Both the protease inhibitory and the antifungal activity were abolished on treatment of API with DTT (5 mM), suggestive of a common site for both the activities. This is the first report on API as a potential biocontrol agent against phytopathogenic fungi. PMID- 10471390 TI - Silica-induced generation of reactive oxygen species in Rat2 fibroblast: role in activation of mitogen-activated protein kinase. AB - Exposure to silica has been associated with progressive pulmonary inflammation and fibrosis. While the fibroblasts play an important role in the pathogenesis of silicosis, the direct interaction between silica and fibroblasts is poorly understood. We observed that silica particles stimulated intracellular ROS generation in Rat2 fibroblast, evidenced by DCFH oxidation. Silica-induced DCFH oxidation was inhibited by catalase and DPI, a flavoenzyme inhibitor. Additionally, the time course of elevation of the intracellular ROS was paralleled by the increases of MEK and ERK phosphorylation. Silica-induced ERK phosphorylation was also effectively attenuated by catalase and DPI. However, SOD enhanced the silica-induced ERK phosphorylation, indicating a role for H(2)O(2) in ERK activation. Furthermore, ERK and MEK phosphorylation are reproduced by H(2)O(2) treatment. Taken together, these results demonstrate that silica stimulates ROS production via flavoenzyme-dependent mechanism in Rat2 fibroblasts and the H(2)O(2), in turn, serves as a signal transduction element in activating MEK-ERK pathway. PMID- 10471391 TI - Laminin stimulates protein tyrosine dephosphorylation in PC12 cells. AB - Laminin stimulates neurite outgrowth in rat pheochromocytoma cells (PC12 cells). Here, we investigated laminin signal transduction mechanisms by adding the tyrosine kinase/phosphatase modulators, genistein, quercetin, aurin tricarboxylic acid (ATA), and vanadate to PC12 cells. At 10 microM both genistein and quercetin enhanced laminin-mediated neurite outgrowth by 1.7- and 2.3-fold, respectively, while at 10 microM, ATA inhibited laminin-mediated neurite outgrowth by 92%. Vanadate inhibited neurite outgrowth by 63% at 10 microM. Immunoblot analysis revealed four proteins of approximately 240, 22, 110, and 35 kDa, which were dephosphorylated on tyrosine residues in laminin-treated PC12 cells, but not in NIH 3T3 cells. These results demonstrate that laminin-mediated neurite outgrowth involves protein tyrosine dephosphorylation and suggests that this mechanism may have specificity to neuronal cells. PMID- 10471392 TI - Malignant transformation of NIH3T3 cells by overexpression of early lymphocyte activation antigen CD98. AB - CD98, which forms a heterodimer of relative molecular mass (M(r)) 125, 000, was originally identified as an early T cell activation antigen. It consists of a heavy chain of M(r) 85,000 that bears the CD98 epitope and a light chain of M(r) 40, 000. CD98 is strongly expressed on the surface of activated lymphocytes and various tumor cells irrespective of tissue origins. To investigate the participation of CD98 in cellular proliferation and malignant transformation, we established and characterized human CD98-transfected NIH3T3 clones. Although the doubling times of the control cells and CD98-transfected clones were almost the same, CD98-transfected clones grew to a higher saturation density than control cells. Effciency of colony formation in soft agar was augmented in CD98 transfected clones, and this augmentation was significantly reduced by anti-human CD98 mAb. Furthermore, CD98-transfected clones developed tumors in athymic mice. These results indicated that overexpression of CD98 participates in the process of malignant transformation, suggesting that CD98 has oncogenic potential. PMID- 10471393 TI - Functional expression of the tandem-repeated homodimer of the mitochondrial ADP/ATP carrier in Saccharomyces cerevisiae. AB - The mitochondrial ADP/ATP carrier (AAC) is believed to function as a dimer. To characterize the oligomeric state of the yeast type 2 AAC (yAAC2), we tried to express its tandem-repeated homodimer, in which the C-terminus of the first repeat was fused to the N-terminus of the second repeat, in yeast mitochondria. The tandem dimer was expressed in the mitochondrial membrane at the same level as that of yAAC2, being inserted into the mitochondrial membrane as in yAAC2, and it showed very similar transport activity to that of yAAC2. It was suggested that the two carrier molecules in a dimeric form are located in the membrane facing each other in the same orientation. PMID- 10471394 TI - Characterization of the VEGF binding site on the Flt-1 receptor. AB - The angiogenic growth factor VEGF binds to the receptor tyrosine kinases Flt-1 and KDR/Flk-1. Immunoglobulin (Ig)-like loop-2 of Flt-1 is involved in binding VEGF, but the contribution of other Flt-1 Ig-loops to VEGF binding remains unclear. We tested the ability of membrane-bound chimeras between the extracellular domain of Flt-1 and the cell adhesion molecule embigin to bind VEGF. VEGF bound as well to receptors containing Flt-1 loops 1-2 or 2-3 as it did to the entire Flt-1 extracellular domain. Chimeras containing only loop-2 of Flt 1 bound VEGF with 22-fold lower affinity. We conclude that high-affinity VEGF binding requires Ig-like loop-2 plus either loop-1 or loop-3. In addition, Flt-1 amino acid residues Arg-224 and Asp-231 were not essential for high-affinity binding of VEGF to membrane-bound Flt-1. PMID- 10471395 TI - Expression of bone morphogenetic protein-2 via adenoviral vector in C2C12 myoblasts induces differentiation into the osteoblast lineage. AB - To examine the effectiveness of a gene transfer of bone morphogenetic protein (BMP)-2 into C2C12 myoblasts, we constructed a human BMP-2-expressing replication deficient adenoviral vector, AxCAOBMP-2. C2C12 cells were infected in vitro with either this viral vector or an Escherichia coli LacZ gene-expressing control adenovirus vector. An efficient gene transfer to the C2C12 cells was confirmed with the LacZ gene-expressing vector by X-gal staining. Abundant BMP-2 expression in C2C12 cells infected with this viral vector was confirmed by immunofluorescence and Western blot analysis. C2C12 cells transferred with the BMP-2 gene by this vector produced alkaline phosphatase in the cells and also produced and secreted osteocalcin in the culture medium, demonstrating that a gene transfer of BMP-2 into C2C12 cells in vitro could convert these cells from myoblast to osteoblast lineage. PMID- 10471396 TI - Caveolin-1 expression and caveolae biogenesis during cell transdifferentiation in lung alveolar epithelial primary cultures. AB - Caveolae are omega-shaped invaginations of the plasmalemma possessing a cytoplasmic membrane protein coat of caveolin. Caveolae are present in the in vivo alveolar epithelial type I (ATI) lung cell, but absent in its progenitor, the alveolar epithelial type II (ATII) cell. In primary culture ATII cells grown on a plastic substratum acquire with time an ATI-"like" phenotype. We demonstrate that freshly isolated rat ATII cells lack caveolae and expression of caveolin-1 (a critical caveolae structural protein). As the ATII cells acquire an ATI-like phenotype in primary culture caveolin-1 expression increases, with caveolin-1 signal at 192 h postseeding up to 50-fold greater than at 60 h; caveolae were morphologically evident only after 132 h. When maintaining the differentiated ATII phenotype with time, i.e., culture upon collagen with an apical interface of air, a temporal increase in caveolin-1 expression was not observed, with only very faint signals evident even at 192 h postseeding; at no time did these cultures display caveolae. In late primary ATII cultures caveolin-1 expression and caveolae biogenesis occur as a function of in vitro transformation from the ATII to the ATI-like phenotype. The results have broad implications for the in vitro study of the role of caveolae and caveolin in alveolar epithelial cell biology. PMID- 10471397 TI - Inhibition of glutathione S-transferase zeta and tyrosine metabolism by dichloroacetate: a potential unifying mechanism for its altered biotransformation and toxicity. AB - Dichloroacetate (DCA) inhibits its own metabolism and is converted to glyoxylate by glutathione S-transferase zeta (GSTz). GSTz is identical to maleylacetoacetate isomerase, an enzyme of tyrosine catabolism that converts maleylacetoacetate (MAA) to fumarylacetoacetate and maleylacetone (MA) to fumarylacetone. MAA and MA are alkylating agents. Rats treated with DCA for up to five days had markedly decreased hepatic GSTz activity and increased urinary excretion of MA. When dialyzed cytosol obtained from human liver was incubated with DCA, GSTz activity was unaffected. In contrast, DCA incubation inhibited enzyme activity in dialyzed hepatic cytosol from rats. Incubation of either rat or human hepatic cytosol with MA led to a dose dependent inhibition of GSTz. These data indicate that humans or rodents exposed to DCA may accumulate MA and/or MAA which inhibit(s) GSTz and, consequently, DCA biotransformation. Moreover, DCA-induced inhibition of tyrosine catabolism may account for the toxicity of this xenobiotic in humans and other species. PMID- 10471398 TI - Mutation "hot spot" in HLA class I-restricted T cell epitope on hepatitis B surface antigen in chronic carriers and hepatocellular carcinoma. AB - Mutations in the human hepatitis B virus (HBV) genome contribute to its escape from host immune surveillance and result in persistent infection. We report the identification of frequent mutations encompassing residues 29 to 53 of the HBV surface antigen in chronic HBV carriers as well as in patients with hepatocellular carcinoma. The location of these mutations, not found in patients with acute hepatitis and vaccinated infants, coincides with a human leukocyte antigen class I-restricted cytotoxic T lymphocyte epitope. Significantly, mutations occur at a higher frequency (83%) compared with those identified on the immunogenic "a" determinant (25%) of the corresponding patients. Our findings therefore suggest the potential importance of this novel mutation "hot spot" in the establishment of chronic HBV infection. PMID- 10471399 TI - Human placental sodium-dependent vitamin C transporter (SVCT2): molecular cloning and transport function. AB - We report here on the cloning and functional characterization of human SVCT2, a sodium-dependent vitamin C (ascorbate) transporter. The hSVCT2 cDNA obtained from a human placental choriocarcinoma cell cDNA library, codes for a protein of 650 amino acids with a predicted molecular mass of 70 kDa. At the level of amino acid sequence, the human SVCT2 exhibits 95% identity to its rat homolog. When functionally expressed in mammalian cells, hSVCT2 induces the transport of ascorbic acid. The transport process induced by hSVCT2 is Na(+)-dependent and is specific for ascorbate. The Michaelis-Menton constant (K(t)) for the transport of ascorbate in cDNA-transfected cells is 69 +/- 5 microM. The relationship between the cDNA-specific uptake rate of ascorbate and Na(+) concentration is sigmoidal with a Na(+):ascorbate stoichiometry of 2:1. Northern blot analysis shows that SVCT2-specific transcripts are present in heart, brain, placenta, and liver and is absent in lung and skeletal muscle. The size of the principal transcript is approximately 7.5 kb. PMID- 10471400 TI - Prevention of quinone-mediated DNA arylation by antioxidants. AB - High performance liquid chromatographic (HPLC) analysis showed that the prototype antioxidant ascorbate (vitamin C) inhibits the DNA adducts induced by synthetic estrogen diethylstilbestrol (DES) and the antiestrogen metabolite 4 hydroxytamoxifen (4-OHTam). Treatment of salmon testes DNA with 4-OHTam quinone or 4-OHTam in the presence of horseradish peroxidase and hydrogen peroxide (H(2)O(2)) generated the same DNA adduct profile. Vitamin C and N-acetylcysteine (NAC) inhibited the formation of 4-OHTam-dG adducts in a dose-dependent manner. To determine whether the same antioxidants also protect cellular DNA, HL-60 cells were used as cell culture model. Cells treated with 10 microM 4-OHTam in the presence of 1 microM H(2)O(2 )for 24 h gave 4-OHTam-dG adducts approximately 4 x 10(-7), n = 3. Treatment of the cells with 100 microM 4-OHTam, without H(2)O(2), produced the same level of adducts. Supplementation of the incubation media with vitamin C (2.5 mM) or NAC (5 mM) inhibited the formation of DNA adducts. Thus, antioxidants may protect susceptible cells from genotoxicity associated with 4 OHTam activation. PMID- 10471401 TI - Gene expressions of opioid receptors and G-proteins in pineal glands. AB - In our previous studies, the opioid receptors located on pinealocytes have been identified and characterized, and these receptors have been found to play a stimulatory role in melatonin synthesis by activating the rate limiting enzyme, N acetyltransferase (NAT). In the present study, by using reverse transcriptase polymerase chain reaction (RT-PCR) followed by nested-PCR, segments of delta and mu opioid receptors have been amplified from mRNA of rat pineal gland and cerebral cortex. In addition, segments of delta and mu opioid receptors have also been amplified from mRNA of human pineal gland. Furthermore, G(alphai/o)- and G(beta)-protein-coupled receptor mRNAs have been amplified and identified from rat pineal gland. The regulatory effects of morphine on G(alphai/o) and G(beta) mRNA levels have been semiquantitatively analyzed. Acute morphine administration caused significant increase in G(alphai/o), and G(beta), mRNA levels in rat pineal gland, but not in other brain regions. Further studies are needed in order to elaborate the mechanisms of these opioid receptors in regulating G-protein expression in pineal gland. PMID- 10471402 TI - Formation and loss of nitrated proteins in peroxynitrite-treated rat skin in vivo. AB - Peroxynitrite is a reactive cytotoxic species, capable of nitrating tyrosine residues to form 3-nitrotyrosine. Little is known about the formation and loss of nitrated proteins in vivo. We have measured nitrated proteins, by enzyme-linked immunosorbent assay, in rat skin after exposure to peroxynitrite. Peroxynitrite (100-200 nmol site(-1)) was injected into the skin of anesthetized rats. At the highest dose 78.6 +/- 9.5 pmol mg(-1) protein of nitrated BSA equivalents were measured at 4 h and a significant increase was observed for 24 h after administration in skin samples. The loss of nitrated proteins from skin appeared biphasic with an initial (t(1/2) = 2 h) and slower loss (t(1/2) = 22 h). A major nitrated protein was identified as albumin by Western blot analysis. The data demonstrate that a single exposure to peroxynitrite can lead to the presence of nitrated proteins in skin for at least 24 h. The sustained presence of nitrated proteins may influence the inflammatory process in skin disease. PMID- 10471403 TI - Capacitation induces tyrosine phosphorylation of proteins in the boar sperm plasma membrane. AB - Capacitation (activation) of mammalian spermatozoa is accompanied by protein phosphorylation, elevation of the intracellular calcium concentration and an increased plasma membrane fluidity. The subcellular localization of tyrosine phosphorylation during capacitation have not yet been elucidated. The aim of this study was to investigate whether boar sperm capacitation induces tyrosine phosphorylation of plasma membrane proteins. Capacitation induced tyrosine phosphorylation of 3 proteins (27, 37, and 40 kDa), which coincided with an increase in the plasma membrane fluidity. The importance of the induced tyrosine phosphorylation in sperm binding to the zona pellucida and the induction of the acrosome reaction is discussed. PMID- 10471404 TI - Identification of a gluconic acid derivative attached to the N-terminus of histidine-tagged proteins expressed in bacteria. AB - Our previous studies have shown that the His tag cleaved from fusion proteins contained two distinct components P1 and P2. P1 has been identified to be a His tagged peptide of G-H-H-H-H-H-H-H-H-H-H-S-S-G-H-I-E-G-R resulted from initiator methionine deletion, and P2 contains an unknown moiety at the second residue glycine of the tag (x-G-H-H-H-H-H-H-H-H-H-H-S-S-G-H-I-E-G-R, x = 178.0 Da). This study aimed to determine the structure of the modification by using a combination of protein isotope labeling and mass spectrometry. His-tagged FKBP was expressed in (15)N and (13)C labeling growth media respectively. Isotopic labeled His tagged proteins ((15)N-His-FKBP and (13)C-His-FKBP) were isolated by affinity chromatography and subjected to Xa digestions to release the labeled His tag. Subsequent analyses of the released His tag by MALDI-TOF-MS indicated a mass difference of 178.0 +/- 0.2 Da, between the two (15)N-labeled peptides P1 and P2, suggesting that the modification moiety contained no nitrogen. A mass difference of 184.0 +/- 0.2 Da was observed on MALDI between (13)C-labeled peptide P1 and P2, indicating six carbons in the modification group. Also, comparing the mass shift on MALDI spectra of P1 and P2 after hydrogen/deuterium exchange revealed that the modification moiety had five hydroxyl groups. It was concluded that the modification was a gluconic acid derivative attached to the N-terminus of His tagged proteins expressed in bacteria. The proposed structure was further confirmed by MALDI analysis of periodate oxidation products of His-tagged peptides. PMID- 10471405 TI - Detection of molecular interactions by using a new peptide-displaying bacteriophage biosensor. AB - Foreign peptides fused to the carboxy terminus of P22 tailspike protein are solvent-exposed and highly antigenic when displayed on the surface of infectious virus particles. Binding of an anti-peptide specific Fab antibody fragment enhances the infectivity of chimeric bacteriophage particles in a titre-dependent fashion. Although the precise molecular basis of this enhanced infectivity remains unclear, experimental data and modelling approaches suggest that the antibody binding might restore conformational impairments in the assembled tail protein affecting its activity and performance during infection. These results suggest that in addition to free enzymes, peptide-displaying bacteriophages could be engineered as new biosensors to detect molecular interactions by using natural viral enzymes critical for cell infection. PMID- 10471406 TI - Cloning and characterization of rat leukotriene B(4) receptor. AB - Leukotriene B(4) (LTB(4)) is a potent chemoattractant for neutrophils and eosinophils. cDNAs for LTB(4) receptor (BLT) have been cloned from human, mouse, and guinea pig. Here we report the isolation of BLT from rat genomic library. Rat BLT consists of 351 amino acids with homologies of 80.2, 93.2, and 71.6%, to human, mouse, and guinea pig BLT, respectively. When expressed in human embryonic kidney (HEK)-293 cells, rat BLT showed a specific and high-affinity binding to LTB(4) with a Kd value of 0.68 nM (mean, n = 3). Northern blot analysis showed that BLT is exclusively expressed in polymorphonuclear leukocytes. Furthermore, the expression of BLT was high in proteosepeptone-activated peritoneal macrophages, while the resident macrophages did not show significant expression. The present results suggest important roles of LTB(4) in macrophage recruitment and activation. PMID- 10471407 TI - Association of peptides with heat shock protein gp96 occurs in vivo and not after cell lysis. AB - Immunization of mice and rats with gp96 preparations isolated from syngeneic cancers has been shown to elicit protective immunity to a number of cancers. The specific immunogenicity of gp96 preparations derives from the antigenic peptides chaperoned by the gp96 molecule and not from gp96 molecules per se. Studies reported here demonstrate that the association of peptides with gp96 occurs in vivo and is not a procedural artifact which occurs in vitro after cell lysis. This demonstration has a bearing on the proposed functional role of HSP peptide association in antigen processing and presentation by MHC I molecules. PMID- 10471408 TI - Allosteric changes of thrombin catalytic site induced by interaction of bothrojaracin with anion-binding exosites I and II. AB - Bothrojaracin, a 27-kDa C-type lectin from Bothrops jararaca venom, is a selective and potent thrombin inhibitor (K(d) = 0.6 nM) which interacts with the two thrombin anion-binding exosites (I and II) but not with its catalytic site. In the present study, we analyzed the allosteric effects produced in the catalytic site by bothrojaracin binding to thrombin exosites. Opposite effects were observed with alpha-thrombin, which possesses both exosites I and II, and with gamma-thrombin, which lacks exosite I. On the one hand, bothrojaracin altered both kinetic parameters K(m) and k(cat) of alpha-thrombin for small synthetic substrates, resulting in an increased efficiency of alpha-thrombin catalytic activity. This effect was similar to that produced by hirugen, a peptide based on the C-terminal hirudin sequence (residues 54-65) which interacts exclusively with exosite I. On the other hand, bothrojaracin decreased the amidolytic activity of gamma-thrombin toward chromogenic substrates, although this effect was observed with higher concentrations of bothrojaracin than those used with alpha-thrombin. In agreement with these observaions, bothrojaracin produced opposite effects on the fluorescence intensity of alpha- and gamma thrombin derivatives labeled at the active site with fluorescein-Phe-Pro-Arg chloromethylketone. These observations support the conclusion that bothrojaracin binding to thrombin produces two different structural changes in its active site, depending on whether it interacts exclusively with exosite II, as seen with gamma thrombin, or with exosite I (or both I and II) as observed with alpha-thrombin. The ability of bothrojaracin to evoke distinct modifications in the thrombin catalytic site environment when interacting with exosites I and II make this molecule an interesting tool for the study of allosteric changes in the thrombin molecule. PMID- 10471409 TI - Phosphorylation and proteasome-dependent degradation of Bcl-2 in mitotic-arrested cells after microtubule damage. AB - Treatment of NIH-OVCAR-3 cells with paclitaxel, a microtubule-stabilizing agent, induces mitotic arrest and apoptosis, but also Bcl-2 phosphorylation. We report here that Bcl-2 phosphorylation precedes Bcl-2 down-regulation and that both events are closely associated with mitotic arrest, but are not sufficient for paclitaxel to trigger apoptosis. Indeed, when paclitaxel-treated cells were induced to exit mitosis in the presence of 2-aminopurine, Bcl-2 phosphorylation and Bcl-2 down-regulation were both inhibited. In contrast, when apoptosis was inhibited by a caspase inhibitor or Bcl-2 over-expression, Bcl-2 phosphorylation and down-regulation still occurred. Furthermore, we show that Bcl-2 is degraded in mitosis by the proteasome-dependent pathway since Bcl-2 down-regulation is inhibited by proteasome inhibitors such as MG132, Lactacystin and LLnL. Taken together these results indicate that mitotic spindle damage results in post translational modifications of Bcl-2 by phosphorylation and degradation. PMID- 10471410 TI - Tomato consumption modulates oxidative DNA damage in humans. AB - Consumption of a single serving of tomatoes by healthy human volunteers was sufficient to alter levels of oxidative DNA base damage in white cell DNA within 24 h. Levels of the mutagenic oxidized purine base 8-hydroxyguanine decreased, especially in those subjects whose initial levels of this base were higher than the mean. However, total DNA base damage remained unchanged since levels of 8 hydroxyadenine rose. The ability of tomato consumption to modulate oxidative DNA damage in the short term may indicate why daily consumption of fruits and vegetables is beneficial in decreasing cancer incidence. PMID- 10471411 TI - Alleic variants of human melatonin 1a receptor: function and prevalence in subjects with circadian rhythm sleep disorders. AB - The human melatonin 1a (hMella) receptor gene was screened for mutations using genomic DNA samples from patients with circadian rhythm sleep disorders and control subjects by single strand conformational polymorphism analysis (SSCP). We found seven mutations, two of which predict amino acid changes R54W and A157V, respectively. The prevalence of the R54W variant and that of the A157V variant were several times more common in non-24-h sleep-wake syndrome subjects than among control subjects, although the incidence was not significant in our study group. When expressed in COS-7 cells, the R54W mutant receptor exhibited significantly reduced B(max) and slightly enhanced affinity (reduced K(d)) compared to the wild type receptor, while the A157V variant receptor showed similar binding characteristics to the wild type. The identification of variants in the hMella receptor will provide a useful tool for analyzing genetic predisposition toward various diseases related to melatonin function and to clarify the physiological role of melatonin receptors in humans. PMID- 10471412 TI - Interferon-alpha-induced inhibition of B16 melanoma cell proliferation: interference with the bFGF autocrine growth circuit. AB - The molecular mechanisms underlying the growth inhibition induced by interferon alpha (IFN-alpha) in B16 murine melanoma cells were investigated. IFN-alpha did not induce cell apoptosis, but strongly interfered with the synthesis of basic fibroblast growth factor (bFGF), which acts as an autocrine growth factor in this system. Inhibition of bFGF synthesis was observed at the same concentrations (50 500 pM, 10-100 U/ml) of IFN-alpha able to induce growth arrest of B16 melanoma cells. Although the synthesis of acidic (a)FGF was only slightly affected by IFN alpha, the cytokine induced release of an aFGF-related low-molecular-weight peptide, which was able to interfere with bFGF binding to surface receptors. Thus, the molecular mechanisms of IFN-alpha activity on melanoma cells include a specific modulation of the bFGF autocrine circuit. PMID- 10471413 TI - Autocrine-paracrine endothelin system in the physiology and pathology of steroid secreting tissues. PMID- 10471414 TI - Neurotransmitter transporters in the central nervous system. PMID- 10471415 TI - Regulation of beta-adrenoceptor signaling in cardiac function and disease. PMID- 10471416 TI - The delta-opioid receptor: molecular pharmacology, signal transduction, and the determination of drug efficacy. PMID- 10471417 TI - Autoregulation of serotonin neurons: role in antidepressant drug action. PMID- 10471418 TI - Carotid endarterectomy : where do we draw the line? PMID- 10471419 TI - The North American Symptomatic Carotid Endarterectomy Trial : surgical results in 1415 patients. AB - BACKGROUND AND PURPOSE: This study reports the surgical results in those patients who underwent carotid endarterectomy in the North American Symptomatic Carotid Endarterectomy Trial (NASCET). METHODS: The rates of perioperative stroke and death at 30 days and the final assessment of stroke severity at 90 days were calculated. Regression modeling was used to identify variables that increased or decreased perioperative risk. Nonoutcome surgical complications were summarized. The durability of carotid endarterectomy was examined. RESULTS: In 1415 patients there were 92 perioperative outcome events, for an overall rate of 6.5%. At 30 days the results were as follows: death, 1.1%; disabling stroke, 1.8%; and nondisabling stroke, 3.7%. At 90 days, because of improvement in the neurological status of patients judged to have been disabled at 30 days, the results were as follows: death, 1.1%; disabling stroke, 0.9%; and nondisabling stroke, 4.5%. Thirty events occurred intraoperatively; 62 were delayed. Most strokes resulted from thromboembolism. Five baseline variables were predictive of increased surgical risk: hemispheric versus retinal transient ischemic attack as the qualifying event, left-sided procedure, contralateral carotid occlusion, ipsilateral ischemic lesion on CT scan, and irregular or ulcerated ipsilateral plaque. History of coronary artery disease with prior cardiac procedure was associated with reduced risk. The risk of perioperative wound complications was 9.3%, and that of cranial nerve injuries was 8.6%; most were of mild severity. At 8 years, the risk of disabling ipsilateral stroke was 5.7%, and that of any ipsilateral stroke was 17.1%. CONCLUSIONS: The overall rate of perioperative stroke and death was 6.5%, but the rate of permanently disabling stroke and death was only 2.0%. Other surgical complications were rarely clinically important. Carotid endarterectomy is a durable procedure. PMID- 10471420 TI - Medical complications associated with carotid endarterectomy. North American Symptomatic Carotid Endarterectomy Trial (NASCET) AB - BACKGROUND AND PURPOSE: Carotid endarterectomy (CE) has been shown to be beneficial in patients with symptomatic high-grade (70% to 99%) internal carotid artery stenosis. To achieve this benefit, complications must be kept to a minimum. Complications not associated with the procedure itself, but related to medical conditions, have received little attention. METHODS: Medical complications that occurred within 30 days after CE were recorded in 1415 patients with symptomatic stenosis (30% to 99%) of the internal carotid artery. They were compared with 1433 patients who received medical care alone. All patients were in the North American Symptomatic Carotid Endarterectomy Trial (NASCET). RESULTS: One hundred fifteen patients (8.1%) had 142 medical complications: 14 (1%) myocardial infarctions, 101 (7.1%) other cardiovascular disorders, 11 (0.8%) respiratory complications, 6 (0.4%) transient confusions, and 10 (0.7%) other complications. Of the 142 complications, 69.7% were of short duration, and only 26.8% prolonged hospitalization. Five patients died: 3 from myocardial infarction and 2 suddenly. Medically treated patients experienced similar complications with one third the frequency. Endarterectomy was approximately 1.5 times more likely to trigger medical complications in patients with a history of myocardial infarction, angina, or hypertension (P<0.05). CONCLUSIONS: Perioperative medical complications were observed in slightly fewer than 1 of every 10 patients who underwent CE. The majority of these complications completely resolved. Most complications were cardiovascular and occurred in patients with 1 or more cardiovascular risk factors. In this selected population, the occurrence of perioperative myocardial infarction was uncommon. PMID- 10471421 TI - Prospective study of aspirin use and risk of stroke in women. AB - BACKGROUND AND PURPOSE: In secondary prevention, aspirin reduces risk of ischemic stroke. In primary prevention of stroke, however, the role of aspirin is uncertain, especially in women. METHODS: In 1980, 79 319 women in the Nurses' Health Study cohort, 34 to 59 years of age and free of diagnosed cardiovascular disease, cancer, and rheumatoid arthritis, completed questionnaires that included information on aspirin use. Data on aspirin use were updated in 1982, 1984, and 1988. By 1994, after 994 231 person-years of follow-up, 503 incident strokes (295 ischemic strokes, 100 subarachnoid hemorrhages, 52 intraparenchymal hemorrhages, and 56 strokes of undetermined type) were documented. RESULTS: There was no clear relationship between aspirin use and risk of total stroke; risk was slightly reduced among women who took 1 to 6 aspirin per week and slightly increased among women who took 7 or more aspirin per week. Women who took 1 to 6 aspirin per week had a lower risk of large-artery occlusive infarction compared with women who reported no aspirin use; after simultaneous adjustment for other cardiovascular risk factors and selected nutrients, the multivariate relative risk was 0.50 (95% CI 0.29 to 0.85, P=0.01). Women who took 15 or more aspirin per week had an excess risk of subarachnoid hemorrhage; the multivariate relative risk was 2.02 (95% CI 1.04 to 3.91, P for trend=0.02). The reduction in large-artery occlusive infarction with aspirin was of greater magnitude for older, hypertensive, or smoking women than for younger, nonhypertensive, or nonsmoking women; the elevation in subarachnoid hemorrhage with aspirin was also more apparent for older or hypertensive women than for younger or nonhypertensive women. Aspirin use was not associated with risk of other subtypes of stroke. CONCLUSIONS: These prospective data indicate that women who take 1 to 6 aspirin per week have a reduced risk of large-artery occlusive infarction, but those who use 15 or more aspirin per week have an increased risk of subarachnoid hemorrhage. This observational study suggests benefits of aspirin for ischemic stroke with low frequency of use and hazards for hemorrhagic stroke with high frequency of use, particularly among older or hypertensive women. Thus, the effect on total stroke will depend on the dose of aspirin and the distribution of stroke subtypes and risk factors in the population. PMID- 10471422 TI - Prospective study of calcium, potassium, and magnesium intake and risk of stroke in women. AB - BACKGROUND AND PURPOSE: High intakes of calcium, potassium, and magnesium have been hypothesized to reduce risks of cardiovascular disease, but only a few prospective studies have examined intakes of these cations in relation to risk of stroke. METHODS: In 1980, 85 764 women in the Nurses' Health Study cohort, aged 34 to 59 years and free of diagnosed cardiovascular disease and cancer, completed dietary questionnaires from which we calculated intakes of calcium, potassium, and magnesium. By 1994, after 1.16 million person-years of follow-up, 690 incident strokes (129 subarachnoid hemorrhages, 74 intraparenchymal hemorrhages, 386 ischemic strokes, and 101 strokes of undetermined type) had been documented. RESULTS: Intakes of calcium, potassium, and magnesium were each inversely associated with age- and smoking-adjusted relative risks of ischemic stroke, excluding embolic infarction of nonatherogenic origin (n=347). Adjustment for other cardiovascular risk factors, including history of hypertension, attenuated these associations, particularly for magnesium intake. In a multivariate analysis, women in the highest quintile of calcium intake had an adjusted relative risk of ischemic stroke of 0.69 (95% CI, 0.50 to 0.95; P for trend=0.03) compared with those in the lowest quintile; for potassium intake the corresponding relative risk was 0.72 (95% CI, 0.51 to 1.01; P for trend=0.10). Further simultaneous adjustment for calcium and potassium intake suggested an independent association for calcium intake. The association of risk with calcium intake did not appear to be log linear; the increase in risk was limited to the lowest quintile of intake, and intakes > approximately 600 mg/d did not appear to reduce risk of stroke further. The inverse association with calcium intake was stronger for dairy than for nondairy calcium intake. Intakes of calcium, potassium, and magnesium were not related to risk of other stroke subtypes. CONCLUSIONS: Low calcium intake, and perhaps low potassium intake, may contribute to increased risk of ischemic stroke in middle-aged American women. It remains possible that women in the lowest quintile of calcium intake had unknown characteristics that made them susceptible to ischemic stroke. PMID- 10471423 TI - Nonfasting serum glucose and insulin concentrations and the risk of stroke. AB - BACKGROUND AND PURPOSE: Type 2 diabetes is an established risk factor for stroke, but the relations between asymptomatic hyperglycemia, hyperinsulinemia, and stroke incidence remain uncertain. We have examined the relationship between established diabetes, nonfasting serum glucose and serum insulin concentrations, and subsequent risk of stroke. METHODS: We performed a prospective study of 7735 men aged 40 to 59 years drawn from general practices in 24 British towns. Men with missing serum glucose values (n=50) and men on insulin injection (n=36) were excluded, leaving 7649 men available for analysis. Baseline nonfasting serum was analyzed for insulin with a specific enzyme-linked immunosorbent assay method in 18 of the 24 towns (n=5663 men). RESULTS: During the mean follow-up period of 16.8 years, there were 347 stroke cases (fatal and nonfatal) in the 7649 men. Men who developed diabetes during follow-up (n=320) and men with established type 2 diabetes at screening (n=98) both showed significantly increased risk of stroke, even after adjustment for cardiovascular risk factors, including blood pressure (adjusted relative risk [RR], 2.27; 95% CI, 1.23 to 4.20; RR, 2.07; 95% CI, 1. 44 to 2.98, respectively). In men with no diagnosed diabetes at screening (n=7551), risk of stroke was increased significantly only in the top 2.5% of the nonfasting glucose distribution (>/=8.2 mmol/L), and this persisted even after adjustment for cardiovascular risk factors, including hypertension (RR, 1.86; 95% CI, 1.11 to 3. 13). Exclusion of the 320 men who developed diabetes during follow-up attenuated this risk so that it was no longer significant (RR, 1.56; 95% CI, 0.83 to 2.91). In the 5567 men with insulin measurements and no diagnosis of diabetes at screening, a J-shaped relationship was seen between nonfasting insulin and risk of stroke. Risk was significantly raised in the first quintile and in the fourth quintile and above compared with the second quintile, with all findings of marginal significance. Part of the increased risk at higher levels of insulin was due to men who developed diabetes in the follow-up period. CONCLUSIONS: This study confirms the importance of established type 2 diabetes as an independent risk factor for stroke. The increased risk of stroke seen in hyperglycemic subjects and those with elevated serum insulin levels at screening reflected to some extent the high proportion of men who subsequently developed diabetes. PMID- 10471424 TI - Proteinuria is an independent risk factor for ischemic stroke in non-insulin dependent diabetes mellitus. AB - BACKGROUND AND PURPOSE: Proteinuria is an independent risk factor for cardiovascular disease in patients with NIDDM. The aim of this study was to assess the relationship between proteinuria and ischemic stroke in subjects with NIDDM, and to determine whether proteinuria is an independent risk factor for stroke. METHODS: We performed a case-control study of 59 diabetic patients with first-ever ischemic stroke due to thrombotic arterial occlusion, who were considered cases, and 180 diabetic patients without stroke, matched by gender, age, and diabetes duration, as a control group. WHO criteria for verified definite or possible stroke were used to ascertain the diagnosis of stroke. For the purpose of this study proteinuria was defined as a 24-hour urinary protein excretion rate of >/=20 and <200 microg/min. Risk factors included were smoking, blood pressure, body mass index, serum total cholesterol, hyperglycemia, and proteinuria. RESULTS: Subjects with stroke had higher proteinuria proportion and systolic and diastolic blood pressures. Both frequency of antihypertensive treatment and antihypertensive drugs used were similar among subjects with and without stroke. In multivariate logistic regression analysis, the ORs and 95% CIs for the variables identified as risk factors for stroke were as follows: systolic pressure (OR 3.10; 95% CI 3.01 to 4.21; P=0.03); diastolic pressure (OR 3.30; 95% CI 1.04 to 4.48; P<0.0001); fasting glucose >/=11.1 mmol (OR 1.82; 905% CI 1.4 to 3.8; P=0.04), HbA1c >/=9.5% (OR 1.7; 95% CI 1.3 to 5.1; P<0.01), and proteinuria (OR 3.23; 95% CI 1.06 to 4.36; P<0.0001). CONCLUSIONS: Our case-control study gives evidence that proteinuria is an independent risk factor for ischemic stroke in patients with NIDDM. PMID- 10471425 TI - Circadian variation in ischemic stroke subtypes. AB - BACKGROUND AND PURPOSE: While previous studies suggest that the peak time period for the occurrence of ischemic stroke is in the mid- to late-morning hours, detailed information pertaining to circadian variations among the various stroke subtypes has been limited. The purpose of our study was to define the circadian patterns of symptom onset in an acute stroke trial with an established system for stroke subtype classification. METHODS: An analysis was conducted on 1272 patients enrolled in the Trial of Org 10172 in Acute Stroke Treatment (TOAST) study. All patients had a documented time of stroke symptom onset, and all stroke subtype determinations were made by a single rater. RESULTS: The Greatest portion of atherothrombotic strokes (25.7%), cardioembolic strokes (30.5%), and strokes of other/unknown mechanism (27.1%) occurred between 6:01 AM and 12:00 noon. The greatest portion of lacunar strokes (31.6%) were present on awakening. More than one half of the infarcts in this series were either present on awakening or occurred in the mid- to late-morning hours. The correlation between stroke subtype and time of symptom onset did not reach statistical significance (P=0.07, Pearson's chi(2) method). CONCLUSIONS: Although there is a trend for clustering of ischemic stroke in the morning hours, there is insufficient specificity to predict with any reasonable likelihood the stroke subtype according to the circadian pattern of symptom onset. PMID- 10471426 TI - Remacemide hydrochloride: a double-blind, placebo-controlled, safety and tolerability study in patients with acute ischemic stroke. AB - BACKGROUND AND PURPOSE: Remacemide hydrochloride and its principal active desglycinyl metabolite are low-affinity noncompetitive N-methyl-D-aspartate (NMDA)-receptor channel blockers. Remacemide hydrochloride has demonstrated neuroprotection in animal models of hypoxia and ischemic stroke. This study assessed the safety, tolerability, and pharmacokinetics of ascending doses of remacemide hydrochloride in patients with recent onset (within 12 hours) ischemic stroke. METHODS: This was a placebo-controlled, dose escalating, parallel group study. Groups of 8 patients (6 active, 2 placebo) were planned to receive twice daily treatment, with l00 mg, 200 mg, 300 mg, 400 mg, 500 mg, or 600 mg remacemide hydrochloride given as 2 intravenous infusions followed by 6 days' oral treatment. Patients who were unable to swallow discontinued study medication but continued to be monitored for safety; these patients were replaced. A CT or MRI scan was performed within 48 hours of admission to establish the cause of focal neurological deficit. Patients with ischemic stroke continued in the study. Patients with other causes of focal neurological deficit were withdrawn and replaced. Because the frequency of dysphagia after stroke in the first dose group (100 mg BID) was higher than had been anticipated, the protocol was amended so that subsequent dose groups received 6 intravenous infusions (2 doses per day for 3 days). Neurological and functional outcome data were collected, but the study was not powered to demonstrate drug efficacy. Patient safety was assessed by clinical observation, laboratory tests, and ECGs, while tolerability was assessed by recording adverse events. Blood sampling was included to determine plasma concentrations of remacemide and the desglycinyl metabolite at fixed points during the dosing period. RESULTS: The most common adverse events considered by the investigator to be possibly treatment related were related to the central nervous system (CNS), and these events appeared to increase with dose. Four patients were withdrawn from the study because of CNS-related events: 1 in the placebo group, 1 in the 500 mg BID group, and 2 in the 600 mg BID group. Infusion site reactions and gastrointestinal upset were also reported and considered to be treatment related. One patient in the placebo group and 4 patients in the 600 mg BID dose group experienced vomiting, whereas this event was not reported by patients in the other dose groups. CONCLUSIONS: On the evidence of this study, the maximum well-tolerated dose for remacemide hydrochloride in acute stroke is 400 mg BID. Doses of 200 mg BID or higher attained the putative neuroprotective plasma concentrations of remacemide predicted from animal models (250 to 600 ng/mL). The expected gradual accumulation of active metabolite might suggest that optimal neuroprotective concentrations are unlikely to be achieved within the early hours of treatment at this dose. However, plasma concentrations do not directly reflect brain concentrations, because studies in rats show that remacemide and the desglycinyl metabolite rapidly reach comparable brain concentrations within 1 hour, despite a lower plasma concentration of the metabolite. PMID- 10471427 TI - Increased cerebral CO(2) reactivity after heparin-mediated extracorporal LDL precipitation (HELP) in patients with coronary heart disease and hyperlipidemia. AB - BACKGROUND AND PURPOSE: There is experimental and clinical evidence that hypercholesterolemia leads to an impairment of endothelial function in coronary and cerebral arteries. Using transcranial Doppler sonography, we examined CO(2) reactivity as a marker of cerebral vasoreactivity in patients with coronary heart disease and hyperlipidemia before and after drastic lowering of LDL cholesterol, lipoprotein(a) [Lp(a)], and fibrinogen levels by heparin-mediated extracorporal LDL precipitation (HELP). METHODS: CO(2) reactivity was determined in 13 patients with coronary artery disease and hyperlipidemia undergoing regular HELP therapy. Middle cerebral artery mean blood flow velocity (MFV) was detected by transcranial Doppler. CO(2) reactivity was calculated as the percent change of MFV during hypercapnia, induced by ventilation of carbogene (5% CO(2), 95% O(2)), to normocapnia. Patients with extracranial or intracranial stenoses were excluded. Other parameters such as blood viscosity, heart rate, and blood pressure were measured to control hemorheologic and systemic influences on CO(2) reactivity. RESULTS: A single HELP treatment reduced total cholesterol, LDL cholesterol, Lp(a), triglycerides, and fibrinogen levels by >50% (P<0.001). Blood viscosity significantly decreased from 1.24+/-0.04 to 1.07+/-0.02 mPa (P<0.001). Blood pressure, heart rate, and MFV did not change significantly. CO(2) reactivity increased from 22% +/- 21% to 36% +/- 18% (P<0.05). CONCLUSIONS: Fast and drastic removal of LDL cholesterol, Lp(a), and fibrinogen from plasma results in an improvement of cerebrovascular reactivity in patients with coronary heart disease and hyperlipidemia. The clinical use of HELP in patients with impaired cerebrovascular reactivity might be promising. PMID- 10471428 TI - Four-dimensional ultrasonographic characterization of plaque surface motion in patients with symptomatic and asymptomatic carotid artery stenosis. AB - BACKGROUND AND PURPOSE: In vitro studies of atherosclerotic plaque fracture mechanics suggest that analysis of local variations in surface deformability may provide information on relative vulnerability to plaque fissuring or rupture. We investigated plaque surface deformations in patients with symptomatic and asymptomatic carotid artery disease using 4-dimensional ultrasonography and techniques for measuring optical flow. METHODS: Four-dimensional ultrasound examinations of carotid artery plaques were performed in 23 asymptomatic and 22 symptomatic patients with 50% to 90% stenosis of the internal carotid artery. Plaque surface motion during 1 cardiac cycle was computed with a hierarchical model-based motion estimator. Results were compared with plaque echogenicity and surface structure. RESULTS: Of the 45 patients examined, plaque surface motion estimates were obtained for 18 asymptomatic and 13 symptomatic patients. There were no significant differences in echogenicity or surface structure of asymptomatic and symptomatic plaques (P>0.05). Results of motion estimation showed that asymptomatic plaques had surface motion vectors of equal orientation and magnitude to those of the internal carotid artery, whereas symptomatic plaques demonstrated evidence of inherent plaque movement. There was no significant difference in maximal plaque velocity between symptomatic and asymptomatic plaques (P<0.14). Maximal discrepant surface velocity (MDSV) in symptomatic plaques was 3.85+/-1.26 mm/s (mean+/-SD), which was significantly higher (P<0.001) than MDSV of asymptomatic plaques with 0.58+/-0.42 mm/s (mean+/ SD). CONCLUSIONS: ++MDSV of carotid artery plaques is significantly different in asymptomatic and symptomatic disease. Further studies are warranted to determine whether plaque surface motion patterns can identify vulnerable plaques in patients with carotid artery stenosis. PMID- 10471429 TI - Clinical application of asymptomatic embolic signal detection in acute stroke: a prospective study. AB - BACKGROUND AND PURPOSE: The detection of asymptomatic embolization with the use of Doppler ultrasound has a number of potential applications in patients with acute stroke. It may provide information on the stroke pathogenesis in individual cases, identify patients with continued embolization, and allow localization of the active embolic source. METHODS: We recruited 119 patients with acute anterior circulation infarction within 72 hours of stroke onset. Transcranial Doppler recordings were possible in 100 (84.0%). Bilateral 1-hour middle cerebral artery (MCA) recordings were made and saved on digital audiotape for blinded offline analysis. When embolic signals were detected during screening of the first recording, simultaneous recording was performed from the ipsilateral MCA and common carotid artery for an additional 30 minutes. In all patients with embolic signals at screening and in matched negative controls, recordings were repeated on days 4, 7, and 14. RESULTS: Embolic signals were detected in the symptomatic MCA in 16 patients (16%). They were more common in patients with carotid stenosis (P<0.0001), occurring in 50% of this group. They were rare in patients with cardioembolic stroke (4.5%) and were not detected in patients with lacunar stroke. In the 16 patients with embolic signals, the proportion with embolic signals fell over time (P=0. 0025), but they were still present in a third at 2 weeks. In 10 patients, localization of the embolic source was possible by simultaneous recording from the MCA and the ipsilateral common carotid artery. CONCLUSIONS: Continued asymptomatic embolization is common after stroke in patients with carotid artery disease and is still present in a significant proportion at 2 weeks. The technique may identify patients at risk of further stroke for more aggressive antiplatelet therapy; this needs to be tested in large prospective studies. The technique may also allow localization of the active embolic source. PMID- 10471430 TI - Contrast-enhanced transcranial color-coded sonography in acute hemispheric brain infarction. AB - PURPOSE: The aim of the present study was to investigate the diagnostic potential of contrast-enhanced transcranial color-coded real-time sonography (CE-TCCS) in otherwise ultrasound-refractory acute stroke patients with an ischemia in the territory of the middle cerebral artery (MCA). Furthermore, correlations of CE TCCS findings with clinical, angiographic, and CT results were investigated. METHODS: In 90 acute stroke patients with inadequate insonation conditions in unenhanced transcranial color-coded real-time sonography (TCCS) examinations, CE TCCS, clinical, angiographic, and CT examinations were performed within 12 hours, 36 hours (CE-TCCS only), and 1 week after onset of clinical symptoms. A CT angiography (CTA) as reference method was available in 39 individuals. After application of a galactose-based echo-enhancing agent, the portion of conclusive ultrasound examinations of the MCA, as manifested by an MCA occlusion, decreased or increased flow velocity (FV), and symmetrical MCA FV, was evaluated. CE-TCCS findings on admission and during follow-up were correlated with infarction size as demonstrated on follow-up CT, and clinical findings were assessed by use of the European Stroke Scale. RESULTS: Adequate diagnosis was achieved in 74 of 90 patients (82%) by the use of echo contrast agents. MCA occlusion or reduction of MCA FV was found in 20 and 27 patients, respectively. MCA occlusion was confirmed by CTA in 17 cases. In one individual, false-positive diagnosis of MCA occlusion was made according to ultrasound criteria. In 5 patients with MCA occlusion, vessel recanalization was observed during follow-up; 15 of 27 patients with decreased flow velocities showed normalization after the third examination that was associated with a significantly better clinical outcome (P<0.0001). Furthermore, MCA occlusion or decreased FV in the first 12 hours were associated with significantly larger infarctions in the MCA territory compared with normal CE-TCCS findings (P<0.0001). CONCLUSIONS: CE-TCCS enables adequate diagnosis in approximately 80% of acute hemispheric stroke patients with insufficient unenhanced TCCS examinations. It is a reliable diagnostic tool regarding MCA mainstem and branch occlusions. Because this method conveys useful information concerning cerebral tissue and clinical prognosis, it may be useful to identify those patients who benefit most from local or intra-arterial thrombolytic therapy. PMID- 10471431 TI - Contrast transcranial Doppler ultrasound in the detection of right-to-left shunts: comparison of different procedures and different contrast agents. AB - BACKGROUND AND PURPOSE: Cardiac right-to-left shunts can be identified by transesophageal echocardiography (TEE) and by transcranial Doppler ultrasound (TCD) with the use of different contrast agents and different provocation procedures. Currently, data on an appropriate time window for the appearance of contrast bubbles in the TCD recording after the injection of the contrast medium and the comparison of different provocation maneuvers to increase right-to-left shunting are insufficient. METHODS: Forty-six patients were investigated by both TEE and bilateral TCD of the middle cerebral artery. The following protocol with 6 injection modes was applied in a randomized way: (1) injection of 10 mL of agitated saline without Valsalva maneuver, (2) injection of 10 mL of agitated saline with Valsalva maneuver, (3) injection of 10 mL of a commercial galactose based contrast agent (Echovist) without Valsalva maneuver, (4) injection of 10 mL of Echovist with Valsalva maneuver, (5) injection of 10 mL of Echovist with standardized Valsalva maneuver, and (6) injection of 10 mL of Echovist with coughing. RESULTS: In 20 patients, a right-to-left shunt was demonstrated by TEE and contrast TCD (shunt-positive). Sixteen patients were negative in both investigations, no patient was positive on TEE and negative on TCD, and 10 patients were only positive on at least 1 TCD investigation but negative during TEE. The amount of microbubbles detected in the various tests decreased in the following order: Echovist and Valsalva maneuver, Echovist with coughing, Echovist and standardized Valsalva maneuver, saline with Valsalva maneuver, Echovist, and saline. With a time window of 20 to 25 seconds for the bubbles to appear in the TCD recording and with a sequence of first Echovist and Valsalva maneuver and then Echovist with coughing, all shunts were reliably identified with a specificity of 65% compared with TEE as the traditional gold standard. The time of first microbubble appearance was not helpful to distinguish between shunts detected on TEE and other shunts. CONCLUSIONS: TCD performed twice with 2 provocation maneuvers using Echovist is a sensitive method to identify cardiac right-to-left shunts also identified by TEE. PMID- 10471433 TI - Evaluation of the American Heart Association Stroke Outcome Classification. AB - BACKGROUND AND PURPOSE: The purpose of this study was to evaluate the concurrent validity of the American Heart Association Stroke Outcome Classification (AHA.SOC) and compare performance of its function classification with that of the Modified Rankin Scale. METHODS: The individuals in this study included the last 105 consecutive subjects who were part of a cohort of 459 stroke patients in the Kansas City Stroke Study. The patients were evaluated with a variety of standardized assessments at enrollment (within 14 days of stroke onset) and followed at 1, 3, and 6 months after stroke. Specifically, we examined validity of AHA.SOC by comparing its 3 domains (ie, Domain, Severe, and Function) with stroke severity. We correlated AHA.SOC-Function with scores of the Barthel Index, Lawton Instrumental Activities of Daily Living (IADL) Scale, and Medical Outcome Study 36-Item Short-Form Health Survey (SF-36) measures of physical function and mental health. Finally, we compared the discriminant ability of AHA.SOC-Function and the Modified Rankin Scale in assessing disability and handicap. These data were analyzed with the use of Spearman rank correlations and Kruskal-Wallis tests. RESULTS: All 3 domains of the AHA.SOC were significantly associated with stroke severity and scores of Barthel Index, Lawton IADL, and SF-36 physical function (all P<0.001). Both AHA.SOC-Function and the Modified Rankin Scale discriminated well the disabilities and handicap measured by Barthel Index, Lawton IADL, and SF-36 physical function (all P<0.001). CONCLUSIONS: The AHA.SOC was able to capture impairments, disabilities, and handicap after stroke. The AHA. SOC-Function performed equally as well as the Modified Rankin Scale in assessing disabilities related to basic activities of daily living but differentiated slightly better than the Modified Rankin Scale in assessing disabilities/handicap related to instrumental activities of daily living. Neither the AHA.SOC-Function nor the Modified Rankin Scale captured differences in mental health after stroke. PMID- 10471432 TI - Early surgical treatment for supratentorial intracerebral hemorrhage: a randomized feasibility study. AB - BACKGROUND AND PURPOSE: The safety and the effectiveness of the surgical treatment of spontaneous intracerebral hemorrhage (ICH) remain controversial. To investigate the feasibility of urgent surgical evacuation of ICH, we conducted a small, randomized feasibility study of early surgical treatment versus current nonoperative management in patients with spontaneous supratentorial ICH. METHODS: Patients with spontaneous supratentorial ICH who presented to 1 university and 2 community hospitals were randomized to surgical treatment or best medical treatment. Principal eligibility criteria were ICH volume >10 cm(3) on baseline CT scan with a focal neurological deficit, Glasgow Coma Scale score >4 at the time of enrollment, randomization and therapy within 24 hours of symptom onset, surgery within 3 hours of randomization, and no evidence for ruptured aneurysm or arteriovenous malformation. The primary end point was the 3-month Glasgow Outcome Scale (GOS). A good outcome was defined as a 3-month GOS score >3. RESULTS: Twenty patients were randomized over 24 months, 9 to surgical intervention and 11 to medical treatment. The median time from onset of symptoms to presentation at the treating hospitals was 3 hours and 17 minutes, the time from randomization to surgery was 1 hour and 20 minutes, and the time from onset of symptoms to surgery was 8 hours and 35 minutes. The likelihood of a good outcome (primary outcome measure: GOS score >3) for the surgical treatment group (56%) did not differ significantly from the medical treatment group (36%). There was no significant difference in mortality at 3 months. Analysis of the secondary 3-month outcome measures showed a nonsignificant trend toward a better outcome in the surgical treatment group versus the medical treatment group for the median GOS, Barthel Index, and Rankin Scale and a significant difference in the National Institutes of Health Stroke Scale score (4 versus 14; P=0.04). CONCLUSIONS: Very early surgical treatment for acute ICH is difficult to achieve but feasible at academic medical centers and community hospitals. The trend toward less 3-month morbidity with surgical intervention in patients with spontaneous supratentorial ICH warrants further investigation of very early clot removal in larger randomized clinical trials. PMID- 10471434 TI - The role of diaschisis in stroke recovery. AB - BACKGROUND AND PURPOSE: Recovery from hemiparesis after stroke has been shown to involve reorganization in motor and premotor cortical areas. However, whether poststroke recovery also depends on changes in remote brain structures, ie, diaschisis, is as yet unresolved. To address this question, we studied regional cerebral blood flow in 7 patients (mean+/-SD age, 54+/-8 years) after their first hemiparetic stroke. METHODS: We analyzed imaging data voxel by voxel using a principal component analysis by which coherent changes in functional networks could be disclosed. Performance was assessed by a motor score and by the finger movement rate during the regional cerebral blood flow measurements. RESULTS: The patients had recovered (P<0. 001) from severe hemiparesis after on average 6 months and were able to perform sequential finger movements with the recovered hand. Regional cerebral blood flow at rest differentiated patients and controls (P<0.05) by a network that was affected by the stroke lesion. During blindfolded performance of sequential finger movements, patients were differentiated from controls (P<0.05) by a recovery-related network and a movement-control network. These networks were spatially incongruent, involving motor, sensory, and visual cortex of both cerebral hemispheres, the basal ganglia, thalamus, and cerebellum. The lesion-affected and recovery-related networks overlapped in the contralesional thalamus and extrastriate occipital cortex. CONCLUSIONS: Motor recovery after hemiparetic brain infarction is subserved by brain structures in locations remote from the stroke lesion. The topographic overlap of the lesion affected and recovery-related networks suggests that diaschisis may play a critical role in stroke recovery. PMID- 10471435 TI - Attenuated stroke severity after prodromal TIA: a role for ischemic tolerance in the brain? AB - BACKGROUND AND PURPOSE: Ischemic tolerance has been extensively studied in experimental models of heart and brain ischemia. While there is some clinical evidence of ischemic tolerance in the heart, it is not known whether the same is true for the human brain. METHODS: We conducted a retrospective case-control study in 148 stroke patients with and without antecedent TIA. RESULTS: Despite no significant differences in baseline characteristics, independence (Rankin scale score of 0 to 1) and favorable outcome (Glasgow Coma Scale score of 5) were significantly associated with prior TIA in univariate analysis. After correction for other cardiovascular risk factors, TIA before stroke also was an independent predictor of mild stroke (Canadian Neurological Scale score of > or= 6.5) in multivariate models (absolute difference 21.6%; P=0.01). CONCLUSIONS: Assuming that a TIA represents an adequate stimulus to elicit ischemic tolerance, our results suggest that ischemic tolerance might occur in the human brain. PMID- 10471436 TI - Non-velocity-related effects of a rigid double-stopped ankle-foot orthosis on gait and lower limb muscle activity of hemiparetic subjects with an equinovarus deformity. AB - BACKGROUND AND PURPOSE: This study investigated the non-velocity-related effects of a 1-bar rigid ankle-foot orthosis on the gait of hemiparetic subjects, with particular emphasis on the muscle activity of the paretic lower limb. METHODS: Twenty-one hemiparetic subjects who had been using an ankle-foot orthosis for equinovarus deformity for <1 week participated. Patients walked cued by a metronome at a comparable speed with and without the orthosis. Dependent variables were basic, limb-dependent cycle parameters, gait symmetry, vertical ground reaction forces, sagittal ankle excursions, and kinesiological electromyogram of several lower limb muscles. RESULTS: The use of the caliper was associated with more dynamic and balanced gait, characterized by longer relative single-stance duration of the paretic lower limb, better swing symmetry, better pivoting over the stationary paretic foot, and better ankle excursions (P<0.05). The functional activity of the paretic quadriceps muscles increased, while the activity of the paretic tibialis anterior muscle decreased (P<0.05). CONCLUSIONS: The orthosis led to a more dynamic and balanced gait, with enhanced functional activation of the hemiparetic vastus lateralis muscle. The study further supports the functional benefits of a rigid ankle-foot orthosis in hemiparetic subjects as an integral part of a comprehensive rehabilitation approach. However, the reduced activity in the tibialis muscle may lead to disuse atrophy and hence long-term dependence on the orthosis. PMID- 10471437 TI - Validation of a standardized assessment of postural control in stroke patients: the Postural Assessment Scale for Stroke Patients (PASS). AB - BACKGROUND AND PURPOSE: Few clinical tools available for assessment of postural abilities are specifically designed for stroke patients. Most have major floor or ceiling effects, and their metrological properties are not always completely known. METHODS: The Postural Assessment Scale for Stroke patients (PASS), adapted from the BL Motor Assessment, was elaborated in concordance with 3 main ideas: (1) the ability to maintain a given posture and to ensure equilibrium in changing position both must be assessed; (2) the scale should be applicable for all patients, even those with very poor postural performance; and (3) it should contain items with increasing difficulty. This new scale has been validated in 70 patients tested on the 30th and 90th days after stroke onset. RESULTS: Normative data obtained in 30 age-matched healthy subjects are presented. The PASS meets the following requirements: (1) good construct validity: high correlation with concomitant Functional Independence Measure (FIM) scores (r=0.73, P=10(-6)), with lower-limb motricity scores (r=0.78, P<10(-6)), and with an instrumental measure of postural stabilization (r=0.48, P<10(-2)); (2) excellent predictive validity: high correlation between PASS scores on the 30th day and FIM scores on the 90th day (r=0.75, P<10(-6)); (3) high internal consistency (Cronbach alpha coefficient=0.95); and (4) high interrater and test-retest reliabilities (average kappa=0.88 and 0.72). CONCLUSIONS: Our results confirm that the PASS is one of the most valid and reliable clinical assessments of postural control in stroke patients during the first 3 months after stroke. PMID- 10471438 TI - Neuropsychological change and S-100 protein release in 130 unselected patients undergoing cardiac surgery. AB - BACKGROUND AND PURPOSE: S-100 protein promises to be a valuable surrogate end point for cerebral injury. This is of particular interest within the context of cardiac surgery. We sought to explore the relationship between change in neurospychological performance attributable to cardiopulmonary bypass and the release of brain-specific S-100 protein. METHODS: In an observational comparative study in a University Hospital Cardiac Surgical Unit, S-100 protein release during and 5 hours after the onset of cardiopulmonary bypass was compared with change (from preoperative to 6 to 8 weeks postoperative) in neuropsychological tests in 130 patients undergoing the full range of cardiac surgical procedures. RESULTS: Neuropsychological performance usually improved, being significantly so in 10 of 25 parameters. S-100 area under the curve (AUC) protein release correlated with age (r=0.24, P<0.008) and bypass time (r=0.17, P<0.02). S-100 Cmax correlated with bypass times (r=0.29, P<0.0001). Bypass times correlated with memory performance (Rey R5; r=-0.21, P<0.03). Less S-100 protein release was associated with better neuropsychological performance, as indexed by significant correlations with the Rey Auditory Verbal Learning memory test, descending Critical Flicker Fusion thresholds, and the Hospital Anxiety and Depression rating scales, typically around r=0.2. Multiple regression models showed that neuropsychological tests accounted for 23% of the variance associated with S-100 AUC release, after partialing out the effects of age and bypass time. CONCLUSIONS: The correlation between S-100 protein release and neuropsychological function supports the belief that it is a measure of brain injury, which may be useful in future studies of mechanisms and prevention. PMID- 10471439 TI - Poststroke depression correlates with cognitive impairment and neurological deficits. AB - BACKGROUND AND PURPOSE: The prevalence of poststroke depression is known to be high, but the knowledge of its neuropsychological correlates is limited. This 12 month prospective study was designed to evaluate the natural history of poststroke depression and to study its neuropsychological, clinical, and functional associates. METHODS: We studied a series of 106 consecutive patients (46 women and 60 men, mean age 65.8 years) with acute first-ever ischemic stroke. The patients underwent a neurological, psychiatric, and neuropsychological examination at 3 and 12 months after the stroke. The psychiatric diagnosis of depression was based on DSM-III-R-criteria. RESULTS: Depression was diagnosed in 53% of the patients at 3 months and in 42% of the patients at 12 months after the stroke. The prevalence of major depression was 9% at 3 months and 16% at 12 months. There was an association between poststroke depression and cognitive impairment; the domains most likely to be defective in stroke-related depression were memory (P=0.022), nonverbal problem solving (P=0.039), and attention and psychomotor speed (P=0.020). The presence of dysphasia increased the risk of major depression. The depressive patients were more dependent in ADL and had more severe impairment and handicap than the nondepressive patients. CONCLUSIONS: More than half of the patients suffer from depression after stroke, and the frequency of major depression seems to increase during the first year. In addition to dysphasia, poststroke depression is correlated with other cognitive deficits. We emphasize the importance of psychiatric evaluation of stroke patients. PMID- 10471440 TI - Evaluation of genetic risk factors for silent brain infarction. AB - BACKGROUND AND PURPOSE: Silent brain infarction (SBI) is often found with white matter hyperintensity. A recent genetic study on elderly twins indicated that the susceptibility to white matter hyperintensity was largely determined by genetic factors, implying the existence of genetic susceptibility for SBI as well. We therefore studied 3 genetic polymorphisms in SBI, the deletion/insertion polymorphism of angiotensin-converting enzyme (ACE) gene, the apolipoprotein(a) [apo(a)] size polymorphism, and the T677C polymorphism of methylenetetrahydrofolate reductase (MTHFR) gene, by a case-control study. METHODS: By MRI, 147 subjects with SBI and 214 without cerebral infarctions (control group) were selected from participants of a health examination of the brain. Seventy-four patients with symptomatic subcortical infarction (SSI) from the same area were also included in the study. In addition to the control group, 2 more reference populations were recruited. Typing of the apo(a) size polymorphism was done by Western blotting with the use of an anti-apo(a) antibody. Genotypes of ACE and MTHFR were determined by polymerase chain reaction amplification of the genomic DNA and subsequent restriction enzyme digestion. RESULTS: The ACE polymorphism was not associated with either SBI or SSI. In contrast, the small apo(a) was associated with both SSI and SBI. The MTHFR polymorphism was associated only with SSI. The association of MTHFR and apo(a) was greater in the younger subjects. CONCLUSIONS: Among the 3 genetic polymorphisms studied, only the apo(a) size polymorphism is a risk factor for SBI. PMID- 10471441 TI - Mutations in the COL5A1 coding sequence are not common in patients with spontaneous cervical artery dissections. AB - BACKGROUND AND PURPOSE: The dermal connective tissue of most patients with spontaneous cervical artery dissections (sCAD) contains abnormal collagen fibers. This suggests a predisposing connective tissue defect. The ultrastructural abnormalities in the skin of patients with sCAD have similarity with the morphological alterations in patients with Ehlers-Danlos syndrome type II, a dominant hereditary disorder that has been correlated in some patients to mutations within the genes encoding type V collagen. The aim of this study was to assess the alpha 1 chain of type V collagen (COL5A1) as a candidate gene for sCAD. METHODS: We searched for mutations in the COL5A1 gene in cDNA from cultured fibroblasts of 19 patients with sCAD using single-strand conformational polymorphism analysis and nucleotide sequence analysis of polymerase chain reaction-amplified fragments of the whole COL5A1 coding sequence. RESULTS: We detected 1 missense mutation leading to a predicted amino acid (192D/N) substitution within the N-terminal propeptide in 2 siblings. All other patients showed regular COL5A1 sequences with some silent polymorphisms. CONCLUSIONS: Mutations in the COL5A1 gene do not appear to be a major factor in the etiology of sCAD. PMID- 10471442 TI - Combination drug therapy and mild hypothermia: a promising treatment strategy for reversible, focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: Hypothermia has been suggested to be the most potent therapeutic approach to reduce experimental ischemic brain injury identified to date, and mild hypothermia is increasingly used for neuroprotection during neurovascular surgery. We have recently demonstrated that combined administration of tirilazad mesylate and magnesium provides for an overall enhanced neuroprotective effect. The present study was designed to determine whether the efficacy of mild hypothermia (33 degrees C) can be increased by combination pharmacotherapy with tirilazad and magnesium (MgCl(2)). METHODS: Forty Sprague Dawley rats were subjected to transient, middle cerebral artery occlusion and were randomly assigned to 1 of 4 treatment arms (n=10 each): (1) normothermia+vehicle, (2) normothermia+tirilazad+MgCl(2), (3) hypothermia+vehicle, or (4) hypothermia+tirilazad+MgCl(2). Cortical blood flow was monitored by a bilateral laser-Doppler flowmeter, and the electroencephalogram was continuously recorded. Functional deficits were quantified by daily neurological examinations. Infarct volume was assessed after 7 days. RESULTS: Tirilazad+MgCl(2), hypothermia, and hypothermia+tirilazad+MgCl(2) reduced total infarct volume by 56%, 63%, and 77%, respectively, relative to controls. In animals treated with both hypothermia and combination pharmacotherapy, cortical infarction was almost completely abolished (-99%), and infarct volume in the basal ganglia was significantly reduced by 55%. In addition, this treatment provided for the best electrophysiological recovery and functional outcome. CONCLUSIONS: The neuroprotective efficacy of hypothermia can be increased by pharmacological antagonism of excitatory amino acids and free radicals by using clinically available drugs. This treatment strategy could be of great benefit when applied during temporary artery occlusion in cerebrovascular surgery. PMID- 10471443 TI - Expression of cyclooxygenase-2 mRNA after global ischemia is regulated by AMPA receptors and glucocorticoids. AB - BACKGROUND AND PURPOSE: Cyclooxygenase-2 (COX-2) is implicated in ischemic neuronal death. In focal ischemia, its mRNA induction is mediated through N methyl-D-aspartic acid (NMDA) receptors and phospholipase A(2). Because mechanisms of neuronal death involving COX-2 in global ischemia are unclear, we studied the time course and regulation of COX-2 expression in rat brain global ischemia. METHODS: Global ischemia was induced by a 4-vessel occlusion method. COX-2 mRNA levels were demonstrated with in situ hybridization and COX-2 protein with immunocytochemistry. Several animals were pretreated with MK-801, an NMDA receptor antagonist; 2, 3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX), an alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist; and dexamethasone. RESULTS: In the cortex, the CA3 hippocampal region and dentate gyrus expression of COX-2 mRNA peaked at 4 to 8 hours, while in the CA1 region COX-2 mRNA levels were high at 4 to 24 hours. COX 2 protein was induced in the corresponding regions at 12 to 24 hours, but in the CA1 neurons the protein was still seen at 3 days. COX-2 mRNA induction in the cortex was inhibited by NBQX and dexamethasone and in CA1 neurons was inhibited by NBQX. MK-801 did not suppress COX-2 induction. CONCLUSIONS: COX-2 is differentially induced in the cortex and hippocampal structures after global ischemia. The prolonged COX-2 expression in the vulnerable CA1 neurons is regulated by AMPA receptors, suggesting that COX-2 expression is likely to be associated with AMPA receptor-mediated neuronal death in global ischemia. Glucocorticoids may not be efficiently used to inhibit ischemia-induced COX-2 expression in the hippocampus. PMID- 10471445 TI - Inflammatory responses in the cerebral cortex after ischemia in the P7 neonatal Rat. AB - BACKGROUND AND PURPOSE: The contribution of inflammatory response to the pathogenesis of ischemic lesions in the neonate is still uncertain. This study described the chronological sequence of inflammatory changes that follow cerebral ischemia with reperfusion in the neonatal P7 rat. METHODS: P7 rats underwent left middle cerebral artery electrocoagulation associated with 1-hour left common carotid artery occlusion. The spatiotemporal pattern of cellular responses was characterized immunocytochemically with the use of antibodies against rat endogenous immunoglobulins to visualize the area of the breakdown of the blood brain barrier. Infiltration of neutrophils and T lymphocytes was demonstrated by antibodies against myeloperoxidase and a pan-T cell marker, respectively. Antibodies ED1 and OX-42 were applied to identify microglial cells and macrophages. The response of astrocytes was shown with antibodies against glial fibrillary acidic protein. Cell survival was assessed by Bcl-2 expression. Cell death was demonstrated by DNA fragmentation with the use of the terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling (TUNEL) assay and Bax immunodetection. RESULTS: Endogenous immunoglobulin extravasation through the blood-brain barrier occurred at 2 hours of recirculation and persisted until 1 month after ischemia. Neutrophil infiltration began at 24 hours and peaked at 72 to 96 hours (30+/-3.4 neutrophils per 0.3 mm(2); P<0.0001), then disappeared at 14 days after ischemia. T cells were observed between 24 and 96 hours of reperfusion. Resident microglia-macrophages exhibited morphological remnants and expressed the cell death inhibitor Bcl-2 at 24 hours of recirculation. They became numerous within the next 48 hours and peaked at 7 days after ischemia. Phenotypic changes of resident astrocytes were apparent at 24 hours, and they proliferated between 48 hours and 7 days after ischemia. Progressively inflammatory cells showed DNA fragmentation and the cell death activator Bax expression. Cell elimination continued until there was a complete disappearance of the frontoparietal cortex. CONCLUSIONS: These data demonstrate that perinatal ischemia with reperfusion triggers acute inflammatory responses with granulocytic cell infiltration, which may be involved in accelerating the destructive processes. PMID- 10471444 TI - 1,4-Dihydropyridine calcium channel blockers inhibit plasma and LDL oxidation and formation of oxidation-specific epitopes in the arterial wall and prolong survival in stroke-prone spontaneously hypertensive rats. AB - BACKGROUND AND PURPOSE: Calcium-channel blockers (CCBs) reduce systolic blood pressure and stroke-related mortality in stroke-prone spontaneously hypertensive rats (SPSHR). Brain ischemia is associated with loss of intracellular antioxidants. Increased formation of oxygen radicals and oxidation of LDL may enhance arterial vasoconstriction by various mechanisms. CCBs that also exert antioxidative properties in vitro may therefore be particularly useful. To investigate such antioxidant effects in vivo, we determined several parameters of LDL oxidation in SPSHR treated with two 1,4-dihydropyridine-type (1,4-DHP) CCBs of different lipophilic properties and compared them with antioxidant-treated and untreated controls. We also tested whether these drugs decrease the formation of oxidation-specific epitopes in arteries. METHODS: Five groups of 9 to 14 SPSHR each (aged 8 weeks) were treated with 80 mg/kg body wt per day nifedipine, 1 mg or 0.3 mg/kg body wt per day lacidipine, vitamin E (100 IU/d), or carrier for 5 weeks. A group of Wistar-Kyoto rats was used as normotensive control. Plasma samples were taken, and LDL was isolated by ultracentrifugation. Then LDL was exposed to oxygen radicals generated by xanthine/xanthine oxidase reaction (2 mmol/L xanthine+100 mU/mL xanthine oxidase), and several parameters of oxidation were determined. The presence of native apolipoprotein B and oxidation-specific epitopes in the carotid and middle cerebral arteries was determined immunocytochemically. RESULTS: 1,4-DHP CCBs completely prevented mortality. Normotensive Wistar-Kyoto rats showed less oxidation than control SPSHR. Plasma lipoperoxide levels were 0.87+/-0.27 micromol/L in control SPSHR, 0.69+/-0.19 and 0.63+/-0.20 micromol/L in the groups treated with 0.3 and 1 mg lacidipine, respectively, and 0.68+/-0.23 micromol/L in nifedipine-treated animals (P<0.05 versus control SPSHR for all values). Both CCBs significantly decreased formation of conjugated dienes and prolonged the lag time in LDL exposed to oxygen radicals. Similarly, lipoperoxides and malondialdehyde were significantly reduced (P<0.05). Reduced relative electrophoretic mobility and increased trinitrobenzenesulfonic acid reactivity of LDL from treated rats (P<0.01) also indicated that fewer lysine residues of apolipoprotein B were oxidatively modified in the presence of 1,4-DHP CCBs. Finally, these drugs reduced the intimal presence of apolipoprotein B and oxidized LDL (oxidation-specific epitopes) in carotid and middle cerebral arteries. CONCLUSIONS: In the SPSHR model, 1,4-DHP CCBs reduce plasma and LDL oxidation and formation of oxidation specific epitopes and prolong survival independently of blood pressure modifications. Our results support the concept that the in vivo protective effect of these drugs on cerebral ischemia and stroke may in part result from inhibition of oxidative processes. PMID- 10471446 TI - Adenovirus-mediated kallikrein gene delivery reduces aortic thickening and stroke induced death rate in Dahl salt-sensitive rats. AB - BACKGROUND AND PURPOSE: Kallikrein gene delivery has been shown to attenuate hypertension, cardiac hypertrophy, and renal injury in hypertensive animal models. The aim of this study was to investigate the potential protective effects of kallikrein gene delivery in salt-induced stroke and cerebrovascular disorders. METHODS: Adenovirus harboring the human tissue kallikrein gene (AdCMV-cHK) was delivered intravenously into Dahl salt-sensitive (DS) rats after 4 weeks of high salt loading, and blood pressure was monitored weekly for 9 weeks. RESULTS: A single injection of AdCMV-cHK caused a significant reduction of systolic blood pressure compared with that in control rats, with or without an injection of adenovirus carrying the LacZ (control) gene (AdCMV-LacZ). A maximal blood pressure reduction of 21 mm Hg was observed 2 weeks after gene delivery. The stroke mortality rate of DS rats (AdCMV-LacZ group versus the AdCMV-cHK group) was significantly decreased: 38% versus 9% at 3 weeks and 54% versus 27% at 5 weeks after gene delivery. Kallikrein gene delivery significantly attenuated salt induced aortic hypertrophy, as evidenced by reduced thickness of the aortic wall. Recombinant human tissue kallikrein was detected in rat serum and urine after gene transfer. Kinin-releasing activities in the brain as well as urinary kinin and cGMP levels were significantly increased in rats receiving the kallikrein gene. CONCLUSIONS: This is the first study to demonstrate the protective effect of kallikrein gene delivery in reducing salt-induced stroke mortality and vascular dysfunction. PMID- 10471447 TI - Evidence for selective effects of chronic hypertension on cerebral artery vasodilatation to protease-activated receptor-2 activation. AB - BACKGROUND AND PURPOSE: Protease-activated receptor-2 (PAR-2) can be activated after proteolysis of the amino terminal of the receptor by trypsin or by synthetic peptides with a sequence corresponding to the endogenous tethered ligand exposed by trypsin (eg, SLIGRL-NH(2)). PAR-2 mediates nitric oxide (NO) dependent dilatation in cerebral arteries, but it is unknown whether PAR-2 function is altered in cardiovascular diseases. Since hypertension selectively impairs NO-mediated cerebral vasodilatation in response to acetylcholine and bradykinin, we sought to determine whether PAR-2-mediated vasodilatation is similarly adversely affected by this disease state. METHODS: We studied basilar artery responses in Wistar-Kyoto rats (WKY) (normotensive) and spontaneously hypertensive rats (SHR) in vivo (cranial window preparation) and in vitro (isolated arterial rings). The vasodilator effects of acetylcholine, sodium nitroprusside, and activators of PAR-2 and protease-activated receptor-1 (PAR-1) were compared in WKY versus SHR. Immunohistochemical localization of PAR-2 was also assessed in the basilar artery. RESULTS: Increases in basilar artery diameter in response to acetylcholine were 65% to 85% smaller in SHR versus WKY, whereas responses to sodium nitroprusside were not different. In contrast to acetylcholine, vasodilatation in vivo to SLIGRL-NH(2) was largely preserved in SHR, and SLIGRL-NH(2) was approximately 3-fold more potent in causing vasorelaxation in SHR versus WKY in vitro. In both strains, responses to SLIGRL NH(2) were abolished by N(G)-nitro-L-arginine, an inhibitor of NO synthesis. Activators of PAR-1 had little or no effect on the rat basilar artery. PAR-2-like immunoreactivity was observed in both the endothelial and smooth muscle cells of the basilar artery in both strains of rat. CONCLUSIONS: These data indicate that NO-mediated vasodilatation to PAR-2 activation is selectively preserved or augmented in SHR and may suggest protective roles for PAR-2 in the cerebral circulation during chronic hypertension. PMID- 10471448 TI - Cerebral blood flow during hemodilution and hypoxia in rats : role of ATP sensitive potassium channels. AB - BACKGROUND AND PURPOSE: Hypoxia and hemodilution both reduce arterial oxygen content (CaO(2)) and increase cerebral blood flow (CBF), but the mechanisms by which hemodilution increases CBF are largely unknown. ATP-sensitive potassium (K(ATP)) channels are activated by intravascular hypoxia, and contribute to hypoxia-mediated cerebrovasodilatation. Although CaO(2) can be reduced to equal levels by hypoxia or hemodilution, intravascular PO(2) is reduced only during hypoxia. We therefore tested the hypothesis that K(ATP) channels would be unlikely to contribute to cerebrovasodilatation during hemodilution. METHODS: Glibenclamide (19.8 microg) or vehicle was injected into the cisterna magna of barbiturate-anesthetized rats. The dose of glibenclamide was chosen to yield an estimated CSF concentration of 10(-4) M. Thirty minutes later, some animals underwent either progressive isovolumic hemodilution or hypoxia (over 30 minutes) to achieve a CaO(2) of approximately 7.5 mL O(2)/dL. Other animals did not undergo hypoxia or hemodilution and served as controls. Six groups of animals were studied: control/vehicle (n=4), control/glibenclamide (n=4), hemodilution/vehicle (n=10), hemodilution/glibenclamide (n=10), hypoxia/vehicle (n=10), and hypoxia/glibenclamide (n=10). CBF was then measured with (3)H nicotine in the forebrain, cerebellum, and brain stem. RESULTS: In control/vehicle rats, CBF ranged from 72 mL. 100 g(-1). min(-1) in forebrain to 88 mL. 100 g(-1) x min(-1) in the brain stem. Glibenclamide treatment of control animals did not influence CBF in any brain area. Hemodilution increased CBF in all brain areas, with flows ranging from 128 mL. 100 g(-1) x min(-1) in forebrain to 169 mL. 100 g(-1) x min(-1) in the brain stem. Glibenclamide treatment of hemodiluted animals did not affect CBF in any brain area. Hypoxia resulted in a greater CBF than did hemodilution, ranging from 172 mL. 100 g(-1) x min(-1) in forebrain to 259 mL. 100 g(-1) x min(-1) in the brain stem. Glibenclamide treatment of hypoxic animals significantly reduced CBF in all brain areas (P<0.05). CONCLUSIONS: Both hypoxia and hemodilution increased CBF. Glibenclamide treatment significantly attenuated the CBF increase during hypoxia but not after hemodilution. This finding supports our hypothesis that K(ATP) channels do not contribute to increasing CBF during hemodilution. Because intravascular PO(2) is normal during hemodilution, this finding supports the hypothesis that intravascular PO(2) is an important regulator of cerebral vascular tone and exerts its effect in part by activation of K(ATP) channels in the cerebral circulation. PMID- 10471449 TI - Fluoxetine dilates isolated small cerebral arteries of rats and attenuates constrictions to serotonin, norepinephrine, and a voltage-dependent Ca(2+) channel opener. AB - BACKGROUND AND PURPOSE: Recent clinical observations question that the antidepressant effect of fluoxetine (Prozac) can be explained solely with serotonin reuptake inhibition in the central nervous system. We hypothesized that fluoxetine affects the tone of vessels and thereby modulates cerebral blood flow. METHODS: A small branch of rat anterior cerebral artery (195+/-15 microm in diameter at 80 mm Hg perfusion pressure) was isolated, cannulated, and pressurized (at 80 mm Hg), and changes in diameter were measured by videomicroscopy. RESULTS: Fluoxetine dilated small cerebral arteries with an EC(50) of 7.7+/-1.0x10(-6) mol/L, a response that was not affected by removal of the endothelium or application of 4-aminopyridine (an inhibitor of aminopyridine sensitive K(+) channels), glibenclamide (an inhibitor of ATP-sensitive K(+) channels), or tetraethylammonium (a nonspecific inhibitor of K(+) channels). The presence of fluoxetine (10(-6) to 3x10(-5) mol/L) significantly attenuated constrictions to serotonin (10(-9) to 10(-5) mol/L) and norepinephrine (10(-9) to 10(-5) mol/L). Increasing concentrations of Bay K 8644 (a voltage-dependent Ca(2+) channel opener, 10(-10) to 10(-6) mol/L) elicited constrictions, which were markedly reduced by 2x10(-6) and 10(-5) mol/L fluoxetine, whereas 3x10(-5) mol/L fluoxetine practically abolished the responses. CONCLUSIONS: Fluoxetine elicits substantial dilation of isolated small cerebral arteries, a response that is not mediated by endothelium-derived dilator factors or activation of K(+) channels. The finding that fluoxetine inhibits constrictor responses to Ca(2+) channel opener, as well as serotonin and norepinephrine, suggests that fluoxetine interferes with the Ca(2+) signaling mechanisms in the vascular smooth muscle. We speculate that fluoxetine increases cerebral blood flow in vivo, which contributes to its previously described beneficial actions in the treatment of mental disorders. PMID- 10471450 TI - Effects of S-nitrosoglutathione on acute vasoconstriction and glutamate release after subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Subarachnoid hemorrhage (SAH) causes acute vasoconstriction that contributes to ischemic brain injury shortly after the initial bleed. It has been theorized that decreased availability of nitric oxide (NO) may contribute to acute vasoconstriction. Therefore we examined the effect of the NO donor N-nitroso glutathione (GSNO) on acute vasoconstriction and early ischemic glutamate release after experimental SAH. METHODS: SAH was induced by the endovascular suture method in anesthetized rats. GSNO (1 micromol/L/kg, n=31) or saline (n=21) was injected 5 minutes after SAH. Sham-operated rats received GSNO (1 micromol/L/kg, n=5) 5 minutes after sham surgery. Arterial and intracranial pressures, cerebral blood flow (CBF), and extracellular glutamate release were measured serially for 60 minutes after SAH. SAH size was determined, and vascular measurements were made histologically. RESULTS: GSNO had no effect on resting blood pressure, intracranial pressure, cerebral perfusion pressure, or CBF in sham-operated animals. However, administration of GSNO after SAH was associated with significantly increased CBF (161.6+/-26.6% versus saline 37.1+/ 5.5%, 60 minutes after SAH, P<0.05), increased blood vessel diameter (internal carotid artery [ICA] 285.0+/-16.5 microm versus saline 149.2+/-14.1 microm, P<0.01), decreased vessel wall thickness (ICA12.9+/-0.7 microm versus saline 25.1+/-1.6 microm, P<0.01), and decreased extracellular glutamate levels (3315.6+/-1048.3% versus saline469. 7+/-134.3%, P<0.05). Blood pressure decreased transiently, whereas intracranial pressure, cerebral perfusion pressure, and SAH size were not affected. CONCLUSIONS: These results suggest that GSNO can reverse acute vasoconstriction and prevent ischemic brain injury after SAH. This further implies that acute vasoconstriction contributes significantly to ischemic brain injury after SAH and is mediated in part by decreased availability of NO. PMID- 10471451 TI - Exacerbation of delayed cell injury after transient global ischemia in mutant mice with CuZn superoxide dismutase deficiency. AB - BACKGROUND AND PURPOSE: We have demonstrated that copper-zinc superoxide dismutase (CuZn-SOD), a cytosolic isoenzyme of SODs, has a protective role in the pathogenesis of superoxide radical-mediated brain injury. Using mice bearing a disruption of the CuZn-SOD gene (Sod1), the present study was designed to clarify the role of superoxide anion in the pathogenesis of selective vulnerability after transient global ischemia. METHODS: Sod1 knockout homozygous mutant mice (Sod1 -/ ) with a complete absence of endogenous CuZn-SOD activity, heterozygous mutant mice (Sod1 +/-) with a 50% decrease in the activity, and littermate wild-type mice (male, 35 to 45 g) were subjected to global ischemia. Since the plasticity of the posterior communicating artery (PcomA) has been reported to influence the outcome of hippocampal injury, we assessed the relation between the plasticity of PcomAs and the decrease of regional cerebral blood flow in global ischemia. RESULTS: The fluorescence intensity of hydroethidine oxidation, a measurement of ethidium fluorescence for superoxide radicals, was increased in mutant mice 1 day after both 5 and 10 minutes of global ischemia, compared with wild-type mice. Hippocampal injury in the PcomA hypoplastic brains showed significant exacerbation in mutant mice compared with wild-type littermates 3 days after 5 minutes of global ischemia, although a marked difference was not observed at 1 day. CONCLUSIONS: These data suggest that superoxide radicals play an important role in the pathogenesis of delayed injury in the vulnerable hippocampal CA1 subregion after transient global ischemia. PMID- 10471452 TI - Neuroprotective properties of statins in cerebral ischemia and stroke. AB - BACKGROUND: The atheroma-retarding properties of beta-hydroxy-beta-methylglutaryl coenzyme A reductase (HMG-CoA) inhibitors, or "statins," in both the coronary and carotid arterial beds are well established. However, a growing body of recent data suggests that statins possess important adjunctive properties that may confer additional benefit beyond the retardation of atherosclerosis. In this article, we review the emerging evidence that statins have beneficial effects within the cerebral circulation and brain parenchyma during ischemic stroke and reperfusion. SUMMARY OF REVIEW: Clinical studies show that statins reduce the incidence of ischemic stroke through probable effects on precerebral atherosclerotic plaque and through antithrombotic mechanisms. Additionally, statins have been shown to reduce infarct size in experimental animal models of stroke. Statins both upregulate endothelial nitric oxide synthase (eNOS) and inhibit inducible nitric oxide synthase (iNOS), effects that are potentially neuroprotective. The preservation of eNOS activity in cerebral vasculature, particularly in the ischemic penumbra, may be especially important in preserving blood flow and limiting neurological loss. Statins may also attenuate the inflammatory cytokine responses that accompany cerebral ischemia, and they possess antioxidant properties that likely ameliorate ischemic oxidative stress in the brain. CONCLUSIONS: In addition to reducing stroke, the statin class of drugs exhibits a number of important neuroprotective properties that likely attenuate the effects of ischemia on the brain vasculature and parenchyma. Further investigation of the role of statins in human neuroprotection by use of neuroimaging and cognitive studies is warranted to explore these preliminary observations. In addition to reducing ischemic stroke, early evidence indicates that statins may also be neuroprotective. PMID- 10471453 TI - A standardized MRI stroke protocol: comparison with CT in hyperacute intracerebral hemorrhage. PMID- 10471454 TI - Abstracts of literature PMID- 10471455 TI - Preventing ischemic stroke in patients with prior stroke and transient ischemic attack : a statement for healthcare professionals from the Stroke Council of the American Heart Association. PMID- 10471456 TI - The effect of spironolactone on morbidity and mortality in patients with severe heart failure. Randomized Aldactone Evaluation Study Investigators. AB - BACKGROUND AND METHODS: Aldosterone is important in the pathophysiology of heart failure. In a doubleblind study, we enrolled 1663 patients who had severe heart failure and a left ventricular ejection fraction of no more than 35 percent and who were being treated with an angiotensin-converting-enzyme inhibitor, a loop diuretic, and in most cases digoxin. A total of 822 patients were randomly assigned to receive 25 mg of spironolactone daily, and 841 to receive placebo. The primary end point was death from all causes. RESULTS: The trial was discontinued early, after a mean follow-up period of 24 months, because an interim analysis determined that spironolactone was efficacious. There were 386 deaths in the placebo group (46 percent) and 284 in the spironolactone group (35 percent; relative risk of death, 0.70; 95 percent confidence interval, 0.60 to 0.82; P<0.001). This 30 percent reduction in the risk of death among patients in the spironolactone group was attributed to a lower risk of both death from progressive heart failure and sudden death from cardiac causes. The frequency of hospitalization for worsening heart failure was 35 percent lower in the spironolactone group than in the placebo group (relative risk of hospitalization, 0.65; 95 percent confidence interval, 0.54 to 0.77; P<0.001). In addition, patients who received spironolactone had a significant improvement in the symptoms of heart failure, as assessed on the basis of the New York Heart Association functional class (P<0.001). Gynecomastia or breast pain was reported in 10 percent of men who were treated with spironolactone, as compared with 1 percent of men in the placebo group (P<0.001). The incidence of serious hyperkalemia was minimal in both groups of patients. CONCLUSIONS: Blockade of aldosterone receptors by spironolactone, in addition to standard therapy, substantially reduces the risk of both morbidity and death among patients with severe heart failure. PMID- 10471457 TI - A population-based study of the clinical expression of the hemochromatosis gene. AB - BACKGROUND AND METHODS: Hereditary hemochromatosis is associated with homozygosity for the C282Y mutation in the hemochromatosis (HFE) gene on chromosome 6, elevated serum transferrin saturation, and excess iron deposits throughout the body. To assess the prevalence and clinical expression of the HFE gene, we conducted a population-based study in Busselton, Australia. In 1994, we obtained blood samples for the determination of serum transferrin saturation and ferritin levels and the presence or absence of the C282Y mutation and the H63D mutation (which may contribute to increased hepatic iron levels) in 3011 unrelated white adults. We evaluated all subjects who had persistently elevated transferrin-saturation values (45 percent or higher) or were homozygous for the C282Y mutation. We recommended liver biopsy for subjects with serum ferritin levels of 300 ng per milliliter or higher. The subjects were followed for up to four years. RESULTS: Sixteen of the subjects (0.5 percent) were homozygous for the C282Y mutation, and 424 (14.1 percent) were heterozygous. The serum transferrin saturation was 45 percent or higher in 15 of the 16 who were homozygous; in 1 subject it was 43 percent. Four of the homozygous subjects had previously been given a diagnosis of hemochromatosis, and 12 had not. Seven of these 12 patients had elevated serum ferritin levels in 1994; 6 of the 7 had further increases in 1998, and 1 had a decrease, although the value remained elevated. The serum ferritin levels in the four other homozygous patients remained in the normal range. Eleven of the 16 homozygous subjects underwent liver biopsy; 3 had hepatic fibrosis, and 1, who had a history of excessive alcohol consumption, had cirrhosis and mild microvesicular steatosis. Eight of the 16 homozygous subjects had clinical findings that were consistent with the presence of hereditary hemochromatosis, such as hepatomegaly, skin pigmentation, and arthritis. CONCLUSIONS: In a population of white adults of northern European ancestry, 0.5 percent were homozygous for the C282Y mutation in the HFE gene. However, only half of those who were homozygous had clinical features of hemochromatosis, and one quarter had serum ferritin levels that remained normal over a four-year period. PMID- 10471459 TI - Hypoglycemia due to an insulin-secreting small-cell carcinoma of the cervix. PMID- 10471458 TI - Hereditary hemochromatosis in adults without pathogenic mutations in the hemochromatosis gene. AB - BACKGROUND AND METHODS: Hereditary hemochromatosis in adults is usually characterized by mutations in the HFE gene on the short arm of chromosome 6. Most patients have a substitution of tyrosine for cysteine at position 282 (C282Y). We studied a large family from Italy that includes persons who have a hereditary iron-overload condition indistinguishable from hemochromatosis but without apparent pathogenic mutations in the HFE gene. We performed biochemical, histologic, and genetic studies of 53 living members of the family, including microsatellite analysis of chromosome 6 and direct sequencing of the HFE gene. RESULTS: Of the 53 family members, 15 had abnormal serum ferritin levels, values for transferrin saturation that were higher than 50 percent, or both. Thirteen of the 15 had elevated body iron levels, diagnosed on the basis of the clinical evaluation and liver biopsy, and underwent iron-removal therapy. The other two, both children, did not undergo liver biopsy or iron-removal therapy. None of the 15 members had the C282Y mutation of the HFE gene; 5 of the 15 (as well as 5 healthy relatives) had another mutation of this gene, a substitution of aspartate for histidine at position 63, but none were homozygous for it. No other mutations were found after sequencing of the entire HFE gene for all family members. Microsatellite analysis showed no linkage of the hemochromatosis phenotype with the short arm of chromosome 6, the site of the HFE gene. CONCLUSIONS: Hereditary hemochromatosis can occur in adults who do not have pathogenic mutations in the hemochromatosis gene. PMID- 10471460 TI - Images in clinical medicine. Emphysematous pyelonephritis. PMID- 10471461 TI - Cutaneous wound healing. PMID- 10471462 TI - Clinical problem-solving. Back to the basics. PMID- 10471463 TI - The Journal and its owner--resolving the crisis. PMID- 10471464 TI - Aldosterone and spironolactone in heart failure. PMID- 10471465 TI - Clinical implications of the hemochromatosis gene. PMID- 10471466 TI - Tumor-induced hypoglycemia. PMID- 10471467 TI - Apoptosis and heart failure. PMID- 10471468 TI - Correction: Treatments for Wasting in Patients with the Acquired Immunodeficiency Syndrome. PMID- 10471470 TI - Receptor-mediated transport of foreign DNA into preimplantation mammalian embryos. AB - Mouse and rabbit preimplantation embryos with intact zona pellucida were incubated for 3 hr with DNA-carrying constructs containing insulin as an internalizable ligand: (insulin-polylysine)-DNA and (insulin-polylysine)-DNA (streptavidin-polylysine)-(biotinylated adenovirus). Video-intensified microscopy demonstrated that the constructs penetrated the zona pellucida and accumulated in the blastomere perinuclear space. The percentage of blastocysts formed was about 70% after incubation of zygotes and two-cell embryos with the constructs. Foreign DNA was detected after 51 hr in 80% of rabbit embryos and after 96 hr in 73% of mouse embryos. Inclusion of various adenoviruses into the construct improved foreign DNA preservation in early embryos. Blot hybridization revealed genome integrated foreign DNA in 12- and 15-day mouse embryos and in a newborn. Thus, the ligand-mediated mechanism can be employed for introducing foreign genetic material into early mammalian embryos; insulin provides for delivery inside the cell and to the nucleus, while adenoviruses ensure release from endosomes. PMID- 10471469 TI - Neuronal apoptosis inhibitory protein (NAIP) may enhance the survival of granulosa cells thus indirectly affecting oocyte survival. AB - In mammals, the postnatal loss of more than 99% of female germ cells is due mainly to the process of ovarian follicular atresia. Atresia is known to be mediated by apoptotic granulosa cell-death. Here we show the involvement of neuronal apoptosis inhibitory protein (NAIP) in ovarian folliculogenesis in which it prevents granulosa cell-death. NAIP has been isolated in association with a neurodegenerative disorder, spinal muscular atrophy (SMA), in which it has been shown to suppress mammalian cell-death induced by a variety of stimuli (Liston et al., 1996, Nature 379:349-353). In an in situ hybridization analysis with mouse ovaries, active expression of NAIP mRNA was localized in the granulosa cells of developing follicles from the primary stage to the Graafian stages, whereas the NAIP gene was not expressed or was weakly expressed in follicles that might be undergoing atresia. Gonadotropin, which is known to inhibit apoptosis in ovarian follicles, caused a 2.4-fold increase in NAIP gene expression in the ovary. To study the role of ovarian NAIP, antisense NAIP oligonucleotides were locally delivered into the ovarian bursa. Suppression of ovarian NAIP expression with antisense oligonucleotides evoked a decrease in the number of morphologically normal ovulated oocytes, implying an indirect involvement of NAIP in germ cell development by enhancing the survival of granulosa cells. These findings suggest that gonadotropin-inducible NAIP may indirectly affect oocyte survival as a result of the inhibition of apoptotic granulosa cell-death during folliculogenesis. PMID- 10471471 TI - Expression of milk protein genes is induced directly by exogenous STAT5 without prolactin-mediated signal transduction in transgenic mice. AB - STAT5 is a member of the family of STAT (signal transducer for activating transcription), which was isolated from nuclear extracts of the lactating gland of sheep. It was reported that, in vitro, STAT5 could be activated directly by prolactin (PRL) without PRL-mediated signal transduction. The present study was conducted to investigate above the possibility in vivo, using the transgenic mice overexpressing ovine STAT5 (oSTAT5) under the control of MMTV promoter. Three transgenic mouse lines that expressed the exogenously introduced oSTAT5 in their mammary glands were established. The expression levels of exogenous oSTAT5 were higher than those of endogenous STAT in the mammary glands of all of the three transgenic lines. Although the expression levels of endogenous milk protein genes, WAP, and beta-casein genes, were not correlated with oSTAT5 expression, the expression levels of WAP and beta-casein were induced by exogenous oSTAT5 in the transgenic mice. The present study demonstrated, at very least, that STAT5 expression can directly activate the expression of milk protein genes, and particularly the WAP gene without PRL-mediated signal transduction. PMID- 10471472 TI - Stage-specific effects of defined mixtures of polychlorinated biphenyls on in vitro development of rabbit preimplantation embryos. AB - PCBs adversely affect various reproductive functions. Little is known about the embryo- toxic effects during the preimplantation period in mammals. In the present study the effects of various mixtures of highly purified PCB-congeners on embryo morphology, blastocyst formation, embryo size and cell proliferation were investigated. For 24 hr, day 3 morulae and day 4 blastocysts were cultured in the presence/absence of coplanar congeners (PCB77, PCB126, and PCB169), non-coplanar congeners (PCB28, PCB52, PCB101, PCB118, PCB138, PCB153, PCB180) or both mixtures in concentrations ranging from 0.3 ng/mL to 60 microg/mL total PCB. The main effects were (1) degeneration of all embryos at 60 microg/mL, (2) reduction of cell proliferation in day 4 embryos only by coplanar PCB; in day 3 embryos, however, by all PCB-mixtures, and (3) reduction of cell proliferation in a non linear dose response with the strongest impairment caused by the lowest concentration. Cell proliferation was decreased by 0.3 ng/mL coplanar PCB to 50% of the level in control blastocysts. Our results show that purified PCB congeners in the range of 0.3 ng/mL to 30 microg/mL affect the development of preimplantation embryos in a stage-specific and congener-specific manner. This study provides first evidence for an embryotoxic potential of coplanar PCB congeners. PMID- 10471473 TI - Regulation of cell adhesion during embryonic compaction of mammalian embryos: roles for PKC and beta-catenin. AB - Beta-catenin has a number of roles in early development including involvement in cell adhesion, cell signaling, and developmental fate specification. This study investigates the mechanisms that regulate embryonic compaction, the first cell adhesion event in early mammalian development. Mammalian embryos can be induced to compact at an earlier developmental stage than normal by treatment with agonists that activate protein kinase C (PKC), and this treatment is used to identify and analyze the minimum essential changes required for embryonic compaction. It was predicted that: (1) since activation of PKC can induce compaction prematurely in mouse embryos, phosphorylation of the protein components of the adherens complex would occur during induced compaction and that these components would be required for the cell adhesive event; (2) these same proteins should be phosphorylated during compaction in normal development; (3) new, highly-specific inhibitors of PKC activity would inhibit compaction during normal development and induced compaction; and (4) some PKC isotypes would become localized to the junctional membranes during the process of compaction. In agreement with these predictionst, beta-catenin became phosphorylated on serine/threonine residues both during induced compaction and normal development. Inhibitors to PKC, but not inhibitors to other kinases, blocked compaction. Furthermore, the alpha isotype of PKC is recruited to the membranes of the apposing blastomeres both during induced compaction and during normal development immediately before compaction begins and before beta-catenin becomes part of the detergent-resistant cytoskeleton at the junction. PMID- 10471474 TI - Purification and characterization of fertility-associated antigen (FAA) in bovine seminal fluid. AB - Heparin-binding proteins (HBP) recognized by a monoclonal antibody (M1) are produced by male accessory sex glands and bind to distinct regions of ejaculated bull sperm. Immunoblots of sperm proteins probed with M1 identified HBP variants of approximately 31-, 24-, and 21.5-kDa that were associated with increased fertility of bulls. The purpose of this study was to identify the 31-kDa HBP known as fertility-associated antigen (FAA). FAA was isolated by heparin-affinity chromatography and reversed-phase high performance liquid chromatography near homogeneity. Biochemical characterization indicated that FAA was an unglycosylated, basic protein. FAA protein was detected in seminal vesicle and prostate gland homogenates, and FAA extracted from sperm membranes by treatment with hypertonic media was identical biochemically to seminal fluid-derived FAA. N terminal sequence analysis of purified FAA yielded a 26 amino acid sequence (L K I X S F N V R S F G E S K K A G F N A M R V I V) with 73% identity to a recently identified human deoxyribonuclease (DNase) I-like protein. Two internal amino acid sequences generated from lys-C digested FAA were 85% and 92% identical to the same DNase I-like protein. In conclusion, we have identified a bovine seminal heparin-binding protein that binds to sperm and is indicative of bull fertility as being similar to the family of DNase I-like proteins. These data demonstrate the presence of a novel DNase I-like protein in bull accessory sex glands and form the groundwork for the identification of a candidate genetic marker for fertility of bulls. PMID- 10471475 TI - Involvement of estrogens in the process of sex differentiation in two fish species: the rainbow trout (Oncorhynchus mykiss) and a tilapia (Oreochromis niloticus). AB - In order to study the physiological implication of sex steroid hormones in gonadal sex differentiation in fish, we first investigated the potential role of estrogens using two fish models: the rainbow trout (Oncorhynchus mykiss) and a tilapia species (Oreochromis niloticus). All experiments were carried out on genetically all-male (XY) and all-female (XX) populations. In vivo treatments with an aromatase inhibitor (ATD, 1,4,6- androstatriene-3-17-dione) result in 100% masculinization of an all-female population in rainbow trout (dosage 50 mg/kg of food) and 75.3% in tilapia (dosage 150 mg/kg of food). In tilapia, the effectiveness of the aromatase inhibition by ATD is demonstrated by the marked decrease of the gonadal aromatase activity in treated animals versus control. No masculinization is obtained following treatment with an estrogen receptor antagonist (tamoxifen) in both species. Aromatase and estrogen receptor gene expression was studied in rainbow trout by semi-quantitative RT-PCR in gonads sampled before, during and after sex-differentiation. Aromatase mRNA is specifically detected in female gonads, 3 weeks before the first sign of histological sex-differentiation, i.e., first female meiosis. Aromatase expression in male gonads is at least a few hundred times less than in female gonads. Estrogen receptor gene is expressed in both male and female gonads at all stages with no dimorphic expression between sexes. Specific aromatase gene expression before ovarian differentiation was also demonstrated using virtual Northern blot, with no expression detected in male differentiating gonads. From these results it can be concluded that estrogen synthesis is crucial for ovarian differentiation, and transcription of the aromatase gene can be proposed as a key step in that process in fish. PMID- 10471476 TI - Three-dimensional environments preserve extracellular matrix compartments of ovarian follicles and increase FSH-dependent growth. AB - In this study we performed a systematic comparative analysis of two culture environments-flat/adhesive liquid and three-dimensional collagen gel-upon in vitro ovarian follicle development. We paid particular attention to the effects of in vitro environments upon the preservation of follicular structure and of peri- and intra-follicular extracellular matrix. We show that flat/adhesive environment leads to an obvious distortion of follicle morphology, marked extracellular matrix modifications and high rates of spontaneous, i.e., FSH independent, follicle disruption. In contrast, three-dimensional collagen gel environments are able to maintain follicular structure with an in vivo-like basal lamina architecture, minimizing spontaneous disruption. Follicle distortions found in flat/adhesive culture systems include a pronounced flattening, causing the follicle horizontal diameters not to adequately reflect follicle volume. Our volume data, based on three-axis follicle diameter measurements, indicate that three-dimensional collagen gel environments increase follicle growth, particularly in response to FSH. This study demonstrates that preservation of both peri- and intra-follicular extracellular matrix compartments during the in vitro growth and differentiation of ovarian follicles is highly desirable, and is now possible through the use of appropriate three-dimensional collagen gel culture environments. This system allows a better understanding of the specific roles played by each of the follicle compartments during development. PMID- 10471477 TI - Inhibition of bovine sperm-oocyte fusion by a monoclonal antibody recognising the TEC-2 epitope on bovine oocytes. AB - The TEC-2 antigenic determinant is a carbohydrate epitope located on a glycoprotein carrier molecule. In the mouse, this epitope is expressed on the zona pellucida and plasma membrane of the oocyte and is associated with the ZP2 glycoprotein and involved in the secondary sperm receptor mechanism. On the bovine oocyte expression is confined to the plasma membrane. The aim of this study was to determine the role the TEC-2 epitope plays during fertilization in the bovine species using the monoclonal antibody TEC-02. Incubating oocytes with the TEC-02 antibody prior to fertilization inhibited cleavage in a dose-dependent manner-the cleavage rate decreased as the concentration of the antibody increased. Significantly more sperm were bound to oocytes exposed to TEC-02 (12 sperm/oocyte) compared to oocytes that were not incubated with the antibody (4 sperm/oocyte). Oocytes treated with the TEC-02 antibody had a 7.5 +/- 3.2% fusion rate and no cortical granule exocytosis compared with oocytes not exposed to the antibody, with 86.5 +/- 5.8% of sperm-oocyte fusions and release of cortical granules. The block to sperm-oocyte fertilization observed in the pretreated group was overcome using intracytoplasmic sperm injection as the method of fertilization that bypassed the fusion process. Although sperm were binding to the oolemma these results suggest that fusion was not occurring and this may be due to the antibody occupying TEC-2 epitope sites involved in the fusion process. In conclusion, the TEC-2 epitope seems to be involved in sperm-oocyte interaction in the bovine species and appears to be involved specifically during the fusion events of fertilization. PMID- 10471478 TI - Inhibition of bovine sperm-oocyte fusion by the carbohydrate GalNAc. AB - The TEC-2 epitope is a carbohydrate located on the plasma membrane (oolemma) of the oocyte and appears to be involved in bovine sperm-oolemma fusion. The carbohydrates N-acetylgalactosamine (GalNAc) and galactose are part of the TEC-2 epitope and this study investigated the involvement of these carbohydrates during bovine fertilization. Gametes were exposed to the carbohydrates GalNAc, galactose, and fructose, and the lectins DBA and Con A to determine whether there was an effect on fertilization. The DBA lectin recognizes the carbohydrate GalNAc, whereas Con A recognizes the carbohydrates glucose and mannose. Oocytes pretreated with the DBA lectin prior to fertilization showed a reduction in cleavage corresponding to an increase in lectin concentrations. There was a significant increase in sperm-oolemma binding although fusion was inhibited. Oocytes exposed to GalNAc prior to sperm insemination had no effect on fertilization, however, sperm pretreatment with the carbohydrate caused inhibition of fertilization, with a reduction in cleavage rates as the GalNAc concentration increased. There was also a significant decrease in sperm-oolemma fusion and a significant increase in sperm-oolemma binding. When gametes were exposed to GalNAc at the time of fertilization a similar response to that seen with sperm pretreatment was observed. The carbohydrates galactose and fructose and the lectin Con A did not affect fertilization. In conclusion, the carbohydrate GalNAc, which is associated with the TEC-2 epitope, has a specific role during bovine sperm-oolemma fusion. This study also suggests that there is a carbohydrate-binding molecule on the sperm that binds GalNAc. PMID- 10471479 TI - Purification, characterization, and molecular cloning of an outer layer protein of carp fertilization envelope. AB - An outer layer protein of carp fertilization envelope (FE), FEO-1, was purified from carp oocytes. The cDNAs encoding FEO-1 were cloned. The mature protein of FEO-1 is 21 kDa in molecular weight and contains 177 amino acid residues whose sequence has 58% identity to the outer layer protein of chick vitelline membrane. In situ hybridization and immunocytochemistry show that FEO-1 is expressed in oocytes and liver. In oocytes, FEO-1 is stored in the cortical granules. During cortical reaction, it is exocytosed to the perivitelline space and then gradually added to the outer layer of FE (FE(o)). FEO-1 first appears as discrete deposits along FE(o), then merges to form a continuous layer. The thickness of FE(o) increases as cortical reaction proceeds. In addition to FEO-1, FE(o) contains cystatin, fibroin-like substance (FLS), and cathepsin-like substance (CLS) as well. They are stored in the cortical granules and are exocytosed to FE(o) simultaneously with FEO-1 during cortical reaction. In FE(o), FEO-1 is present in monomer form and can be completely extracted by sodium dodecyl sulfate (SDS) mercaptoethanol (MSH). On the other hand, the cystatin, FLS, and CLS present in FE(o)are cross-linked together. They are partially extracted by SDS-MSH but can be completely extracted by guanidium thiocyanate-lauroylsarcosine. PMID- 10471480 TI - Responses of monkey epididymal sperm of different maturational status to second messengers mediating protein tyrosine phosphorylation, acrosome reaction, and motility. AB - The maturation of various aspects of sperm function have been demonstrated in monkey and human epididymal sperm, including the ability to undergo the acrosome reaction. The present study aimed to investigate the maturational changes in non human primate sperm in the signal transduction mechanisms leading to the acrosome reaction involving cyclic AMP, Ca(2+) influx, protein kinase C, and protein tyrosine phosphorylation. Sperm from the caput, corpus, and cauda epididymidis of cynomolgus monkeys were incubated in a complete medium for 2.5 hr, followed by 30 min stimulation with 1 mM dibutyryl cAMP and 1 mM caffeine, 50 microM 1, 2 dioctanoyl-sn-glycerol (DOG), and 50 microM Ca(2+)-ionophore A23187. Quantitative Western blotting revealed little difference in tyrosine phosphorylated proteins among the caput, corpus, and cauda sperm without stimulation. Incubation with cAMP increased the amount of tyrosine phosphorylated proteins up to 10-fold in the corpus and cauda sperm, but to a lower extent in the caput sperm. Ca(2+) ionophore attenuated the cAMP stimulation but had no effect on its own. Such responses in tyrosine phosphorylated proteins were in great contrast to the responses in the acrosome reaction, where A23187 was the strongest stimulant, resulting in induction of the reaction in 50 +/- 5%, 11 +/- 5%, and 8 +/- 4% cauda, corpus and caput sperm, respectively (mean +/- sem, n = 6). DOG and cAMP in combination induced acrosome reactions in about 10% of viable cells in the cauda and corpus but not caput sperm. Caput sperm responded to cAMP with increases in percentage motility without forward progression whereas cauda sperm displayed marked kinematic changes expected of hyperactivation. Comparisons of responses suggest that the major tyrosine phosphorylated proteins detected are unlikely to be involved immediately in the precipitation of the acrosome reaction, but more related to flagellar motion. Development of signal transduction pathways is part of the epididymal maturational process. PMID- 10471482 TI - The database and the law. PMID- 10471481 TI - A 140-kDa glycopeptide from the sperm ligand of the vitelline coat of the freshwater bivalve Unio elongatulus, only contains O-linked oligosaccharide chains and mediates sperm-egg interaction. AB - In previous studies we found that sperm binding activity in the vitelline coat of the freshwater bivalve Unio elongatulus is located on the O-linked oligosaccharide chains of gp273, one of the two major components of the extracellular coat, and that fucose plays a key role in this interaction. In this paper we report the partial characterization of a large glycopeptide (about 140 kDa) obtained by cyanogen bromide fragmentation of gp273, that maintains sperm binding activity. Lectin blotting revealed that the glycopeptide reacted with lectins from Arachis hypogaea (PNA) and Lotus tetragonolobus (LTA) but not Canavalia ensiformis (ConA). No other PNA-positive fragments could be detected in the electrophoretic pattern of fragmented gp273 but several ConA-positive fragments of lower molecular weight were present indicating that all the O-linked chains are clustered together in this fragment. Two-dimensional gel electrophoresis of the fragment revealed it to be acidic in nature in contrast with the neutral character of the whole gp273 molecule. Competition binding assay showed that this fragment is a strong inhibitor of the interaction, whereas no effect was detected using the ConA-positive peptides. This confirms that the sperm receptor activity of gp273 is related to its O-linked chains. The immunodominance of this fragment is also discussed. PMID- 10471483 TI - Let sleeping dogs lie? PMID- 10471485 TI - Do you dig my groove? PMID- 10471484 TI - The mojo of methylation. PMID- 10471486 TI - Baby, don't stop! PMID- 10471487 TI - Gadzooks! PMID- 10471488 TI - TOUCHINGbase. PMID- 10471489 TI - Artificial chromosome transgenesis reveals long-distance negative regulation of rag1 in zebrafish. PMID- 10471490 TI - Mutations in GJB6 cause nonsyndromic autosomal dominant deafness at DFNA3 locus. PMID- 10471492 TI - Cardiac conduction defects associate with mutations in SCN5A. PMID- 10471491 TI - Frequent activating mutations of FGFR3 in human bladder and cervix carcinomas. PMID- 10471493 TI - One more imprinting review? PMID- 10471494 TI - A new member of the IL-1 receptor family highly expressed in hippocampus and involved in X-linked mental retardation. AB - We demonstrate here the importance of interleukin signalling pathways in cognitive function and the normal physiology of the CNS. Thorough investigation of an MRX critical region in Xp22.1-21.3 enabled us to identify a new gene expressed in brain that is responsible for a non-specific form of X-linked mental retardation. This gene encodes a 696 amino acid protein that has homology to IL-1 receptor accessory proteins. Non-overlapping deletions and a nonsense mutation in this gene were identified in patients with cognitive impairment only. Its high level of expression in post-natal brain structures involved in the hippocampal memory system suggests a specialized role for this new gene in the physiological processes underlying memory and learning abilities. PMID- 10471495 TI - Irs-2 coordinates Igf-1 receptor-mediated beta-cell development and peripheral insulin signalling. AB - Insulin receptor substrates (Irs proteins) mediate the pleiotropic effects of insulin and Igf-1 (insulin-like growth factor-1), including regulation of glucose homeostasis and cell growth and survival. We intercrossed mice heterozygous for two null alleles (Irs1+/- and Irs2+/-) and investigated growth and glucose metabolism in mice with viable genotypes. Our experiments revealed that Irs-1 and Irs-2 are critical for embryonic and post-natal growth, with Irs-1 having the predominant role. By contrast, both Irs-1 and Irs-2 function in peripheral carbohydrate metabolism, but Irs-2 has the major role in beta-cell development and compensation for peripheral insulin resistance. To establish a role for the Igf-1 receptor in beta-cells, we intercrossed mice heterozygous for null alleles of Igf1r and Irs2. Our results reveal that Igf-1 receptors promote beta-cell development and survival through the Irs-2 signalling pathway. Thus, Irs-2 integrates the effects of insulin in peripheral target tissues with Igf-1 in pancreatic beta-cells to maintain glucose homeostasis. PMID- 10471496 TI - Genome-wide analysis of DNA copy-number changes using cDNA microarrays. AB - Gene amplifications and deletions frequently contribute to tumorigenesis. Characterization of these DNA copy-number changes is important for both the basic understanding of cancer and its diagnosis. Comparative genomic hybridization (CGH) was developed to survey DNA copy-number variations across a whole genome. With CGH, differentially labelled test and reference genomic DNAs are co hybridized to normal metaphase chromosomes, and fluorescence ratios along the length of chromosomes provide a cytogenetic representation of DNA copy-number variation. CGH, however, has a limited ( approximately 20 Mb) mapping resolution, and higher-resolution techniques, such as fluorescence in situ hybridization (FISH), are prohibitively labour-intensive on a genomic scale. Array-based CGH, in which fluorescence ratios at arrayed DNA elements provide a locus-by-locus measure of DNA copy-number variation, represents another means of achieving increased mapping resolution. Published array CGH methods have relied on large genomic clone (for example BAC) array targets and have covered only a small fraction of the human genome. cDNAs representing over 30,000 radiation-hybrid (RH)-mapped human genes provide an alternative and readily available genomic resource for mapping DNA copy-number changes. Although cDNA microarrays have been used extensively to characterize variation in human gene expression, human genomic DNA is a far more complex mixture than the mRNA representation of human cells. Therefore, analysis of DNA copy-number variation using cDNA microarrays would require a sensitivity of detection an order of magnitude greater than has been routinely reported. We describe here a cDNA microarray-based CGH method, and its application to DNA copy-number variation analysis in breast cancer cell lines and tumours. Using this assay, we were able to identify gene amplifications and deletions genome-wide and with high resolution, and compare alterations in DNA copy number and gene expression. PMID- 10471497 TI - Intragenic deletion in the gene encoding ubiquitin carboxy-terminal hydrolase in gad mice. AB - The gracile axonal dystrophy (gad) mouse is an autosomal recessive mutant that shows sensory ataxia at an early stage, followed by motor ataxia at a later stage. Pathologically, the mutant is characterized by 'dying-back' type axonal degeneration and formation of spheroid bodies in nerve terminals. Recent pathological observations have associated brain ageing and neurodegenerative diseases with progressive accumulation of ubiquitinated protein conjugates. In gad mice, accumulation of amyloid beta-protein and ubiquitin-positive deposits occur retrogradely along the sensory and motor nervous systems. We previously reported that the gad mutation was transmitted by a gene on chromosome 5 (refs 10,11). Here we find that the gad mutation is caused by an in-frame deletion including exons 7 and 8 of Uchl1, encoding the ubiquitin carboxy-terminal hydrolase (UCH) isozyme (Uch-l1) selectively expressed in the nervous system and testis. The gad allele encodes a truncated Uch-l1 lacking a segment of 42 amino acids containing a catalytic residue. As Uch-l1 is thought to stimulate protein degradation by generating free monomeric ubiquitin, the gad mutation appears to affect protein turnover. Our data suggest that altered function of the ubiquitin system directly causes neurodegeneration. The gad mouse provides a useful model for investigating human neurodegenerative disorders. PMID- 10471498 TI - Non-type I cystinuria caused by mutations in SLC7A9, encoding a subunit (bo,+AT) of rBAT. AB - Cystinuria (MIM 220100) is a common recessive disorder of renal reabsorption of cystine and dibasic amino acids. Mutations in SLC3A1, encoding rBAT, cause cystinuria type I (ref. 1), but not other types of cystinuria (ref. 2). A gene whose mutation causes non-type I cystinuria has been mapped by linkage analysis to 19q12-13.1 (Refs 3,4). We have identified a new transcript, encoding a protein (bo, +AT, for bo,+ amino acid transporter) belonging to a family of light subunits of amino acid transporters, expressed in kidney, liver, small intestine and placenta, and localized its gene (SLC7A9) to the non-type I cystinuria 19q locus. Co-transfection of bo,+AT and rBAT brings the latter to the plasma membrane, and results in the uptake of L-arginine in COS cells. We have found SLC7A9 mutations in Libyan-Jews, North American, Italian and Spanish non-type I cystinuria patients. The Libyan Jewish patients are homozygous for a founder missense mutation (V170M) that abolishes b o,+AT amino-acid uptake activity when co-transfected with rBAT in COS cells. We identified four missense mutations (G105R, A182T, G195R and G295R) and two frameshift (520insT and 596delTG) mutations in other patients. Our data establish that mutations in SLC7A9 cause non-type I cystinuria, and suggest that bo,+AT is the light subunit of rBAT. PMID- 10471499 TI - MBD2 is a transcriptional repressor belonging to the MeCP1 histone deacetylase complex. AB - Mammalian DNA is methylated at many CpG dinucleotides. The biological consequences of methylation are mediated by a family of methyl-CpG binding proteins. The best characterized family member is MeCP2, a transcriptional repressor that recruits histone deacetylases. Our report concerns MBD2, which can bind methylated DNA in vivo and in vitro and has been reported to actively demethylate DNA (ref. 8). As DNA methylation causes gene silencing, the MBD2 demethylase is a candidate transcriptional activator. Using specific antibodies, however, we find here that MBD2 in HeLa cells is associated with histone deacetylase (HDAC) in the MeCP1 repressor complex. An affinity-purified HDAC1 corepressor complex also contains MBD2, suggesting that MeCP1 corresponds to a fraction of this complex. Exogenous MBD2 represses transcription in a transient assay, and repression can be relieved by the deacetylase inhibitor trichostatin A (TSA; ref. 12). In our hands, MBD2 does not demethylate DNA. Our data suggest that HeLa cells, which lack the known methylation-dependent repressor MeCP2, use an alternative pathway involving MBD2 to silence methylated genes. PMID- 10471500 TI - Mi-2 complex couples DNA methylation to chromatin remodelling and histone deacetylation. AB - Methylation of DNA at the dinucleotide CpG is essential for mammalian development and is correlated with stable transcriptional silencing. This transcriptional silencing has recently been linked at a molecular level to histone deacetylation through the demonstration of a physical association between histone deacetylases and the methyl CpG-binding protein MeCP2 (refs 4,5). We previously purified a histone deacetylase complex from Xenopus laevis egg extracts that consists of six subunits, including an Rpd3-like deacetylase, the RbA p48/p46 histone-binding protein and the nucleosome-stimulated ATPase Mi-2 (ref. 6). Similar species were subsequently isolated from human cell lines, implying functional conservation across evolution. This complex represents the most abundant form of deacetylase in amphibian eggs and cultured mammalian cells. Here we identify the remaining three subunits of this enzyme complex. One of them binds specifically to methylated DNA in vitro and molecular cloning reveals a similarity to a known methyl CpG-binding protein. Our data substantiate the mechanistic link between DNA methylation, histone deacetylation and transcriptional silencing. PMID- 10471501 TI - Selective agenesis of the dorsal pancreas in mice lacking homeobox gene Hlxb9. AB - The initial stages of pancreatic development occur early during mammalian embryogenesis, but the genes governing this process remain largely unknown. The homeodomain protein Pdx1 is expressed in the developing pancreatic anlagen from the approximately 10-somite stage, and mutations in the gene Pdx1 prevent the development of the pancreas. The initial stages of pancreatic development, however, still occur in Pdx1-deficient mice. Hlxb9 (encoding Hb9; ref. 6) is a homeobox gene that in humans has been linked to dominant inherited sacral agenesis and we show here that Hb9 is expressed at early stages of mouse pancreatic development and later in differentiated beta-cells. Hlxb9 has an essential function in the initial stages of pancreatic development. In absence of Hlxb9 expression, the dorsal region of the gut epithelium fails to initiate a pancreatic differentiation program. In contrast, the ventral pancreatic endoderm develops but exhibits a later and more subtle perturbation in beta-cell differentiation and in islet cell organization. Thus, dorsally Hlxb9 is required for specifying the gut epithelium to a pancreatic fate and ventrally for ensuring proper endocrine cell differentiation. PMID- 10471502 TI - Pancreas dorsal lobe agenesis and abnormal islets of Langerhans in Hlxb9 deficient mice. AB - In most mammals the pancreas develops from the foregut endoderm as ventral and dorsal buds. These buds fuse and develop into a complex organ composed of endocrine, exocrine and ductal components. This developmental process depends upon an integrated network of transcription factors. Gene targeting experiments have revealed critical roles for Pdx1, Isl1, Pax4, Pax6 and Nkx2-2 (refs 3,4,5,6,7, 8,9,10). The homeobox gene HLXB9 (encoding HB9) is prominently expressed in adult human pancreas, although its role in pancreas development and function is unknown. To facilitate its study, we isolated the mouse HLXB9 orthologue, Hlxb9. During mouse development, the dorsal and ventral pancreatic buds and mature beta-cells in the islets of Langerhans express Hlxb9. In mice homologous for a null mutation of Hlxb9, the dorsal lobe of the pancreas fails to develop. The remnant Hlxb9-/- pancreas has small islets of Langerhans with reduced numbers of insulin-producing beta-cells. Hlxb9-/- beta-cells express low levels of the glucose transporter Glut2 and homeodomain factor Nkx 6-1. Thus, Hlxb9 is key to normal pancreas development and function. PMID- 10471503 TI - Functions of poly(ADP-ribose) polymerase in controlling telomere length and chromosomal stability. AB - In most eukaryotes, poly(ADP-ribose) polymerase (PARP) recognizes DNA strand interruptions generated in vivo. DNA binding by PARP triggers primarily its own modification by the sequential addition of ADP-ribose units to form polymers; this modification, in turn, causes the release of PARP from DNA ends. Studies on the effects of the disruption of the gene encoding PARP (Adprt1, formerly Adprp) in mice have demonstrated roles for PARP in recovery from DNA damage and in suppressing recombination processes involving DNA ends. Telomeres are the natural termini of chromosomes and are, therefore, potential targets of PARP. Here, by the use of two different techniques, we show that mice lacking PARP display telomere shortening compared with wild-type mice. Telomere shortening is seen in different genetic backgrounds and in different tissues, both from embryos and adult mice. In vitro telomerase activity, however, is not altered in Adprt1-/- mouse fibroblasts. Furthermore, cytogenetic analysis of mouse embryonic fibroblasts reveals that lack of PARP is associated with severe chromosomal instability, characterized by increased frequencies of chromosome fusions and aneuploidy. The absence of PARP does not affect the presence of single-strand overhangs, naturally present at the ends of telomeres. This study therefore reveals an unanticipated role for PARP in telomere length regulation and provides insights into its functions in maintaining genomic integrity. PMID- 10471504 TI - Gross chromosomal rearrangements in Saccharomyces cerevisiae replication and recombination defective mutants. AB - Cancer progression is often associated with the accumulation of gross chromosomal rearrangements (GCRs), such as translocations, deletion of a chromosome arm, interstitial deletions or inversions. In many instances, GCRs inactivate tumour suppressor genes or generate novel fusion proteins that initiate carcinogenesis. The mechanism underlying GCR formation appears to involve interactions between DNA sequences of little or no homology. We previously demonstrated that mutations in the gene encoding the largest subunit of the Saccharomyces cerevisiae single stranded DNA binding protein (RFA1) increase microhomology-mediated GCR formation. To further our understanding of GCR formation, we have developed a novel mutator assay in S. cerevisiae that allows specific detection of such events. In this assay, the rate of GCR formation was increased 600-5, 000-fold by mutations in RFA1, RAD27, MRE11, XRS2 and RAD50, but was minimally affected by mutations in RAD51, RAD54, RAD57, YKU70, YKU80, LIG4 and POL30. Genetic analysis of these mutants suggested that at least three distinct pathways can suppress GCRs: two that suppress microhomology-mediated GCRs (RFA1 and RAD27) and one that suppresses non-homology-mediated GCRs (RAD50/MRE11/XRS2). PMID- 10471505 TI - A radiation hybrid map of the zebrafish genome. AB - Recent large-scale mutagenesis screens have made the zebrafish the first vertebrate organism to allow a forward genetic approach to the discovery of developmental control genes. Mutations can be cloned positionally, or placed on a simple sequence length polymorphism (SSLP) map to match them with mapped candidate genes and expressed sequence tags (ESTs). To facilitate the mapping of candidate genes and to increase the density of markers available for positional cloning, we have created a radiation hybrid (RH) map of the zebrafish genome. This technique is based on somatic cell hybrid lines produced by fusion of lethally irradiated cells of the species of interest with a rodent cell line. Random fragments of the donor chromosomes are integrated into recipient chromosomes or retained as separate minichromosomes. The radiation-induced breakpoints can be used for mapping in a manner analogous to genetic mapping, but at higher resolution and without a need for polymorphism. Genome-wide maps exist for the human, based on three RH panels of different resolutions, as well as for the dog, rat and mouse. For our map of the zebrafish genome, we used an existing RH panel and 1,451 sequence tagged site (STS) markers, including SSLPs, cloned candidate genes and ESTs. Of these, 1,275 (87.9%) have significant linkage to at least one other marker. The fraction of ESTs with significant linkage, which can be used as an estimate of map coverage, is 81.9%. We found the average marker retention frequency to be 18.4%. One cR3000 is equivalent to 61 kb, resulting in a potential resolution of approximately 350 kb. PMID- 10471506 TI - Mitochondrial DNA genotypes in nuclear transfer-derived cloned sheep. AB - Eukaryotic cells contain two distinct genomes. One is located in the nucleus (nDNA) and is transmitted in a mendelian fashion, whereas the other is located in mitochondria (mtDNA) and is transmitted by maternal inheritance. Cloning of mammals typically has been achieved via nuclear transfer, in which a donor somatic cell is fused by electoporation with a recipient enucleated oocyte. During this whole-cell electrofusion, nDNA as well as mtDNA ought to be transferred to the oocyte. Thus, the cloned progeny should harbour mtDNAs from both the donor and recipient cytoplasms, resulting in heteroplasmy. Although the confirmation of nuclear transfer has been established using somatic cell-specific nDNA markers, no similar analysis of the mtDNA genotype has been reported. We report here the origin of the mtDNA in Dolly, the first animal cloned from an established adult somatic cell line, and in nine other nuclear transfer-derived sheep generated from fetal cells. The mtDNA of each of the ten nuclear-transfer sheep was derived exclusively from recipient enucleated oocytes, with no detectable contribution from the respective somatic donor cells. Thus, although these ten sheep are authentic nuclear clones, they are in fact genetic chimaeras, containing somatic cell-derived nuclear DNA but oocyte-derived mtDNA. PMID- 10471507 TI - Mutations in the CCN gene family member WISP3 cause progressive pseudorheumatoid dysplasia. AB - Members of the CCN (for CTGF, cyr61/cef10, nov) gene family encode cysteine-rich secreted proteins with roles in cell growth and differentiation. Cell-specific and tissue-specific differences in the expression and function of different CCN family members suggest they have non-redundant roles. Using a positional candidate approach, we found that mutations in the CCN family member WISP3 are associated with the autosomal recessive skeletal disorder progressive pseudorheumatoid dysplasia (PPD; MIM 208230). PPD is an autosomal recessive disorder that may be initially misdiagnosed as juvenile rheumatoid arthritis. Its population incidence has been estimated at 1 per million in the United Kingdom, but it is likely to be higher in the Middle East and Gulf States. Affected individuals are asymptomatic in early childhood. Signs and symptoms of disease typically develop between three and eight years of age. Clinically and radiographically, patients experience continued cartilage loss and destructive bone changes as they age, in several instances necessitating joint replacement surgery by the third decade of life. Extraskeletal manifestations have not been reported in PPD. Cartilage appears to be the primary affected tissue, and in one patient, a biopsy of the iliac crest revealed abnormal nests of chondrocytes and loss of normal cell columnar organization in growth zones. We have identified nine different WISP3 mutations in unrelated, affected individuals, indicating that the gene is essential for normal post-natal skeletal growth and cartilage homeostasis. PMID- 10471508 TI - Disruption of the glucocorticoid receptor gene in the nervous system results in reduced anxiety. AB - The glucocorticoid receptor (Gr, encoded by the gene Grl1) controls transcription of target genes both directly by interaction with DNA regulatory elements and indirectly by cross-talk with other transcription factors. In response to various stimuli, including stress, glucocorticoids coordinate metabolic, endocrine, immune and nervous system responses and ensure an adequate profile of transcription. In the brain, Gr has been proposed to modulate emotional behaviour, cognitive functions and addictive states. Previously, these aspects were not studied in the absence of functional Gr because inactivation of Grl1 in mice causes lethality at birth (F.T., C.K. and G.S., unpublished data). Therefore, we generated tissue-specific mutations of this gene using the Cre/loxP -recombination system. This allowed us to generate viable adult mice with loss of Gr function in selected tissues. Loss of Gr function in the nervous system impairs hypothalamus-pituitary-adrenal (HPA)-axis regulation, resulting in increased glucocorticoid (GC) levels that lead to symptoms reminiscent of those observed in Cushing syndrome. Conditional mutagenesis of Gr in the nervous system provides genetic evidence for the importance of Gr signalling in emotional behaviour because mutant animals show an impaired behavioural response to stress and display reduced anxiety. PMID- 10471509 TI - A novel member of the F-box/WD40 gene family, encoding dactylin, is disrupted in the mouse dactylaplasia mutant. AB - Early outgrowth of the vertebrate embryonic limb requires signalling by the apical ectodermal ridge (AER) to the progress zone (PZ), which in response proliferates and lays down the pattern of the presumptive limb in a proximal to distal progression. Signals from the PZ maintain the AER until the anlagen for the distal phalanges have been formed. The semidominant mouse mutant dactylaplasia (Dac) disrupts the maintenance of the AER, leading to truncation of distal structures of the developing footplate, or autopod. Adult Dac homozygotes thus lack hands and feet except for malformed single digits, whereas heterozygotes lack phalanges of the three middle digits. Dac resembles the human autosomal dominant split hand/foot malformation (SHFM) diseases. One of these, SHFM3, maps to chromosome 10q24 (Refs 6,7), which is syntenic to the Dac region on chromosome 19, and may disrupt the orthologue of Dac. We report here the positional cloning of Dac and show that it belongs to the F-box/WD40 gene family, which encodes adapters that target specific proteins for destruction by presenting them to the ubiquitination machinery. In conjuction with recent biochemical studies, this report demonstrates the importance of this gene family in vertebrate embryonic development. PMID- 10471510 TI - Ocular albinism: evidence for a defect in an intracellular signal transduction system. AB - G protein-coupled receptors (GPCRs) participate in the most common signal transduction system at the plasma membrane. The wide distribution of heterotrimeric G proteins in the internal membranes suggests that a similar signalling mechanism might also be used at intracellular locations. We provide here structural evidence that the protein product of the ocular albinism type 1 gene (OA1), a pigment cell-specific integral membrane glycoprotein, represents a novel member of the GPCR superfamily and demonstrate that it binds heterotrimeric G proteins. Moreover, we show that OA1 is not found at the plasma membrane, being instead targeted to specialized intracellular organelles, the melanosomes. Our data suggest that OA1 represents the first example of an exclusively intracellular GPCR and support the hypothesis that GPCR-mediated signal transduction systems also operate at the internal membranes in mammalian cells. PMID- 10471513 TI - Autumn's picks. AB - September's harvest includes analytical software, micromanipulators and expandable microtitre plates. PMID- 10471511 TI - Eya1-deficient mice lack ears and kidneys and show abnormal apoptosis of organ primordia. AB - Haploinsufficiency for human EYA1, a homologue of the Drosophila melanogaster gene eyes absent (eya), results in the dominantly inherited disorders branchio oto-renal (BOR) syndrome and branchio-oto (BO) syndrome, which are characterized by craniofacial abnormalities and hearing loss with (BOR) or without (BO) kidney defects. To understand the developmental pathogenesis of organs affected in these syndromes, we inactivated the gene Eya1 in mice. Eya1 heterozygotes show renal abnormalities and a conductive hearing loss similar to BOR syndrome, whereas Eya1 homozygotes lack ears and kidneys due to defective inductive tissue interactions and apoptotic regression of the organ primordia. Inner ear development in Eya1 homozygotes arrests at the otic vesicle stage and all components of the inner ear and specific cranial sensory ganglia fail to form. In the kidney, Eya1 homozygosity results in an absence of ureteric bud outgrowth and a subsequent failure of metanephric induction. Gdnf expression, which is required to direct ureteric bud outgrowth via activation of the c-ret Rtk (refs 5, 6, 7, 8), is not detected in Eya1-/- metanephric mesenchyme. In Eya1-/- ear and kidney development, Six but not Pax expression is Eya1 dependent, similar to a genetic pathway elucidated in the Drosophila eye imaginal disc. Our results indicate that Eya1 controls critical early inductive signalling events involved in ear and kidney formation and integrate Eya1 into the genetic regulatory cascade controlling kidney formation upstream of Gdnf. In addition, our results suggest that an evolutionarily conserved Pax-Eya-Six regulatory hierarchy is used in mammalian ear and kidney development. PMID- 10471512 TI - Globozoospermia in mice lacking the casein kinase II alpha' catalytic subunit. AB - Protein kinase casein kinase II (Ck2) is a cyclic-AMP and calcium-independent serine-threonine kinase that is composed of two catalytic subunits (alpha and alpha') and two regulatory beta-subunits. Ck2 is not a casein kinase in vivo, but over 100 substrates are known. The highly conserved amino acid sequences of its subunits and their broad expression suggest that Ck2 may have a fundamental role in cell function. Ck2 has been implicated in DNA replication, regulation of basal and inducible transcription, translation and control of metabolism. The Ck2alpha and Ck2alpha' isoforms (products of the genes Csnk2a1 and Csnk2a2, respectively) are highly homologous, but the reason for their redundancy and evolutionary conservation is unknown. We find here that Csnk2a2 is preferentially expressed in late stages of spermatogenesis, and male mice in which Csnk2a2 has been disrupted are infertile, with oligospermia and globozoospermia ('round-headed' spermatozoa). This is the first demonstration of a unique role for a Ck2 isoform in development. The primary spermatogenic defect in Csnk2a2-/- testis is a specific abnormality of anterior head shaping of elongating spermatids; this is the first defined gene that regulates sperm head morphogenesis. As the germ cells differentiate, they are capable of undergoing chromatin condensation, although many abnormal cells are deleted through apoptosis or Sertoli cell phagocytosis. The few that survive to populate the epididymis exhibit head abnormalities similar to those described in human globozoospermia, thus Csnk2a2 may be a candidate gene for these inherited syndromes. PMID- 10471514 TI - Secondary change in the substantia nigra induced by incomplete infarct and minor hemorrhage in the basal ganglia due to traumatic middle cerebral arterial dissection. PMID- 10471515 TI - Monitoring the effectiveness of anticoagulative therapy in left atrial spontaneous echo contrast by cerebral microemboli detection. PMID- 10471517 TI - Perimesencephalic hemorrhage. PMID- 10471516 TI - Hemiosteoporosis following stroke: importance of pathophysiologic understanding and histologic evidence. PMID- 10471518 TI - Ischemic stroke and tissue hypodensity on computed tomography PMID- 10471519 TI - Molecular effects of paclitaxel: myths and reality (a critical review). AB - Recent studies on paclitaxel (Taxol), a microtubule-stabilizing agent and effective anti-cancer drug, have identified numerous cellular and molecular effects, such as induction of cytokines and tumor-suppressor genes, indirect cytotoxicity due to secretion of tumor necrosis factor, vast activation of signal transduction pathways and selective activity against cells lacking functional p53. Some of these results, including the immediate activation of signaling pathways and gene expression, have been observed only with paclitaxel concentrations 1,000-fold higher than those required for mitotic arrest and apoptosis. The effects of loss of p53 on paclitaxel cytotoxicity depend on cell type (normal murine fibroblasts vs. human cancer cells) and duration of exposure to paclitaxel; p53 status marginally affects paclitaxel sensitivity in human cancer. Although the biochemistry of mitosis and meiosis has been studied independently of research on the mechanism of action of anti-cancer drugs, it eventually provided insight into the effects of paclitaxel. For example, serine protein phosphorylation, which occurs during mitotic arrest or meiosis, explains paclitaxel-induced hyperphosphorylation of Bcl-2 and Bcl-xL. Although some observations are disputed, such mitotic arrest correlates with paclitaxel cytotoxicity, while there is currently no evidence that any paclitaxel effect at clinically relevant concentrations is independent of its tubulin-binding properties. Thus, paclitaxel exerts two types of effect: mitotic arrest with coincidental serine protein phosphorylation and cytotoxicity at clinically relevant concentrations as well as immediate activation of tyrosine kinase pathways and activation of gene expression at much higher concentrations. PMID- 10471520 TI - Quantitative PCR analysis of c-erb B-2 (HER2/neu) gene amplification and comparison with p185(HER2/neu) protein expression in breast cancer drill biopsies. AB - A PCR assay using capillary electrophoresis was designed for the detection of c erbB-2 gene amplification in alcohol-formalin-acetic acid (AFA)-fixed, paraffin embedded biopsies from 81 consecutive breast tumors. c-erbB-2 expression was analyzed in the same samples using immuno-histochemistry (IHC). In the competitive PCR assay, a single pTag plasmid containing a 4-nucleotide (nt) deleted copy of a 124-nt sequence of c-erbB-2 and a 4-nt-deleted copy of a 120-nt sequence of GAPDH was co-amplified with genomic DNA extracted from 3 10 micrometer-thick tissue sections of the tumor biopsy. The percentage of tumor cells in the biopsy specimen and the percentage of tumor cells stained with the membrane anti-c-erbB-2 monoclonal antibody CB11 were recorded by a single pathologist on 2 consecutive sections. Among 81 consecutive tumor biopsies assayed by PCR, 21 (26%) displayed unequivocal c-erbB-2 amplification (actual gene copy number, AGCN > 4), 47 (58%) displayed no c-erbB-2 amplification (AGCN 0.50). Occupational physical activity was inversely associated with renal cell cancer among men. The absence of association among women might be due to smaller range of exposure, confounding by household work or reproductive factors, or to a difference in biological response to physical activity in men and women. PMID- 10471526 TI - Alterations of p73 preferentially occur in gastric adenocarcinomas with foveolar epithelial phenotype. AB - To establish the possible involvement of p73, a newly discovered p53-related candidate as a tumor-suppressor gene in human stomach carcinogenesis, the allelic status, allele-specific expression and mutations of the gene were investigated using PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, RT-PCR SSCP analysis and direct DNA sequencing in 95 gastric adenocarcinomas. Of these, 32 exhibited the heterozygous p73 allele for the StyI restriction site in exon 2. Among these, the cancer DNA of 12 revealed loss of heterozygosity (LOH) of p73. All of the cancers with p73 LOH exhibited phenotypes of foveolar epithelium of the stomach. RT-PCR SSCP analysis of p73 heterozygous cases demonstrated not only bi-allelic expression of the gene but also relatively reduced expression of the affected allele in 6 of 8 tumors with p73 LOH. No gene mutation was detected in the remaining allele of LOH-positive cancers. Our results suggest that alterations of p73, including LOH and abnormal expression, may play roles in the genesis of foveolar-type gastric adenocarcinomas, though this is not in line with a classical Knudson's "2-hit" model. PMID- 10471527 TI - hMLH1, hMSH2 and hMSH6 mutations in hereditary non-polyposis colorectal cancer families from southern Sweden. AB - We have screened 17 Southern Sweden individuals/families with suspected hereditary non-polyposis colorectal cancer (HNPCC) for mutations in the DNA mismatch repair genes hMLH1, hMSH2 and hMSH6 using denaturing gradient gel electrophoresis, protein truncation test and direct DNA sequencing. The families were selected on the basis of a family history of HNPCC-related tumors or the occurrence of metachronous colorectal cancer/endometrial cancer at young age in an individual with a weak family history of cancer. Furthermore, we required that tumor tissue from at least one individual in the family had to display microsatellite instability. We identified germ-line mutations in 9 individuals from 8 families. Five families had mutations in hMLH1, 4 of which were splice site mutations, 2 had frameshift mutations in hMSH2 and 1 patient with metachronous endometrial and rectal cancer but with a weak family history of cancer had a nonsense mutation in hMSH6. Our results present novel germ-line DNA repair gene mutations, one of these in hMSH6, and demonstrate the diversified mutation spectrum in Sweden, where no founder mutation has so far been identified. PMID- 10471528 TI - Expression of p34(cdc2) and cyclins A and B compared to other proliferative features of non-Hodgkin's lymphomas: a multivariate cluster analysis. AB - In view of recent knowledge on proteins regulating the cell cycle, we re evaluated proliferative features of 98 diffusely growing non-Hodgkin's lymphomas. The combined use of 5 proliferation-associated variables (mitotic indices and percentages of Ki-67(+), p34(cdc2+), cyclin A(+) and cyclin B(+) cells) and their entry into a multivariate cluster analysis separated, without overlaps, the entire cohort into 3 groups (clusters) with (1) low, (2) intermediate and (3) high proliferative activity. Conversely, bivariate plots exposed considerable cluster overlaps. Multivariate stepwise discriminant analysis of all cases revealed a decreasing order of discriminant power for % Ki-67(+) cells > % p34(cdc2+) cells > mitotic index > % cyclin A(+) cells > % cyclin B(+) cells. The combined use of 2 variables only, mitotic index and % p34(cdc2+) cells, allowed a clear-cut separation of clusters 2 and 3. In bivariate plots, correlations were best between % Ki-67(+) cells and % cyclin A(+) cells and between mitotic indices and % cyclin B(+) cells. Except for chronic lymphocytic leukemias, immunocytomas and marginal zone lymphomas (all in cluster 1), individual lymphoma entities were distributed among at least 2 clusters. There was, however, a marked preponderance of mantle cell lymphomas and diffuse follicular center lymphomas in cluster 1 and of diffuse large B-cell lymphomas and peripheral T-cell lymphomas in cluster 2. Anaplastic large-cell lymphomas predominated in cluster 3 and responded best to therapy. PMID- 10471529 TI - Identification of distinct regions of allelic loss on chromosome 13q in nasopharyngeal cancer from paraffin embedded tissues. AB - Our main purpose was to identify tumor suppressor gene loci on chromosome 13 responsible for nasopharyngeal cancer (NPC) development by analyzing loss of heterozygosity (LOH) and RB protein expression in paraffin embedded tissues. Normal and tumor DNA were extracted from microdissected samples, and their whole genomes were amplified using degenerate oligonucleotide primers. The polymerase chain reaction (PCR) products were analyzed by repeated amplification using primers derived from 16 microsatellite regions spanning the long arm of this chromosome. Among 50 informative cases, LOH was observed in 44 tumors. Thirty-one tumors displayed partial loss and provided an informative basis for detailed deletion mapping. Three minimal regions of loss were delineated; the first flanked by D13S120 and D13S219, the second by D13S126 and D13S119, and the third by D13S137 and 13qter. These 3 regions were linked to BRCA2 on 13q12, RB1 on 13q14, and 13q14.3-ter, respectively. Seven and 4 cases showed LOH either on 13q12 or 13q14, respectively. Nineteen cases showed LOH of both loci separately. One NPC displayed 13q12 and 13q14.3-ter LOH. RB protein expression was detectable in 76% of the cases. Ten out of 15 cases with the allelic losses limited to 13q14 showed RB protein expression. Contrasting that, 6 out of 7 cases devoid of RB protein expressions showed 13q14LOH. In conclusion, 13qLOH, involving 3 tumor suppressor gene loci, appears to be a frequent genetic event occurring during NPC development. However, other tumor suppressor genes besides RB1, may be responsible for the majority of 13q14LOH. PMID- 10471530 TI - Idiotypic protein-pulsed dendritic cell vaccination in multiple myeloma. AB - Idiotypic protein (Id) produced by myeloma cells is clone-specific and may be a suitable tumor-specific antigen for immune targeting. Advances in dendritic cell (DC) technology suggest the opportunity for using this potent antigen presentation system to deliver myeloma Id to the autologous host to elicit anti tumor immune responses. We have generated DCs from adherent PBMCs from 6 patients with IgG myeloma. These cells were pulsed with the autologous Id and a control vaccine, KLH, and re-infused i.v. back to the patients on 3 separate occasions. Immune responses to KLH and autologous Id were measured and clinical responses monitored. We found that all treatments were well tolerated without any side effects. All patients developed both B- and T-cell responses to KLH, suggesting the integrity of the host immune system to mount immune responses to an antigen delivered using our vaccination strategy. Id-specific responses were also observed. PBMC proliferative responses to Id were observed in 5 of the 6 patients following treatment. In 2 patients, the responses were associated with the production of IFN-gamma. There were also increases in cytotoxic T-cell precursor frequencies for Id-pulsed autologous targets in 3 patients. B-cell responses characterized by the production of anti-Id IgM occurred in 3 and anti-Id IgG in 4 of the 5 evaluated patients. In 1 patient, a modest (25%) but consistent drop in the serum Id level was observed. Id-pulsed DC vaccination can therefore elicit potentially useful anti-myeloma immune responses in patients with multiple myeloma. PMID- 10471531 TI - Tobacco, alcohol and the risk of gastric cancer by sub-site and histologic type. AB - Few studies have provided information on the role of smoking and alcohol in the carcinogenesis of gastric cancer by sub-site and histologic type. The relationship of snuff dipping with risk of gastric cancer has also been rarely studied. In a population-based case-control study conducted in 5 counties of Sweden from February 1989 to January 1995, a total of 90 cases of gastric cardia cancer, 260 and 164 cases of distal gastric cancer of intestinal and diffuse types, respectively, and 1164 frequency-matched control subjects were personally interviewed about life-time smoking, use of smokeless tobacco and use of alcohol 20 years ago. Current smokers had a higher risk than never-smokers for all 3 kinds of gastric adenocarcinoma [odds ratio (OR) 1.7, 95% confidence interval (CI) 1.0-3.1 for gastric cardia adenocarcinoma; OR 1.8, 95% CI 1.2-2.7 for distal gastric cancer of intestinal type; and OR 2.2, 95% CI 1.4-3.5 for distal gastric cancer of diffuse type], and the risk rose with increasing dose and duration of smoking among current smokers. However, no elevated risk was observed for ex smokers. Neither intake of alcoholic beverages nor snuff dipping was associated with an increased risk of any type of cardia or gastric cancer. Our study did not support the hypothesis that the role of tobacco differs by sub-site and histologic sub-type of gastric cancer. PMID- 10471532 TI - Transcriptional and post-transcriptional mechanisms induce cyclin-D1 over expression in B-chronic lymphoproliferative disorders. AB - Cyclin D1 participates in cell-cycle control, in the progression through the G(1) phase and in the transition from the G(1) to the S phase. The CCND1 locus, located in 11q13, is amplified and cyclin-D1 protein is over-expressed in a wide range of human solid tumors. In some B-lymphoid malignancies, the t(11;14)(q13;q32) translocation joins the Ig heavy-chain locus to the CCND1 locus and leads to cyclin-D1 over-expression. In this study, a series of 127 patients presenting a B-chronic lymphoproliferative disorder (B-CLPD) was analyzed using a competitive RT-PCR designed to detect cyclin-D1-mRNA over-expression. Cyclin-D1 mRNA was expressed in patients with mantle-cell lymphoma (MCL; 10/10), hairy-cell leukemia (HCL; 3/5), B-chronic lymphoid leukemia (B-CLL; 4/111) and B large-cell lymphoma (BLCL; 1/1). Densitometric analysis of RT-PCR products and Western-blot autoradiograms, in addition to cytogenetic data, indicated that activation of the cyclin-D1 gene occurred independently of the t(11;14)(q13;q32) translocation in patients with HCL. Indeed, a normal-sized protein of 36 kDa exhibiting a level incompatible with gene activation by a translocation mechanism was detected in lymphoid cells with a normal karyotype. Moreover, we found a discrepancy between cyclin-D1 mRNA and protein levels in MCL and B-CLL, which suggested that some regulatory mechanisms acting at a post-transcriptional level persist in tumor cells. PMID- 10471533 TI - Selective GRB2 SH2 inhibitors as anti-Ras therapy. AB - Given the key role of Ras in the mitogenic signaling by receptor tyrosine kinases, several targets upstream of Ras may prove to be excellent targets for drugs in the treatment of cancer caused by oncogenic tyrosine kinases. CGP78850 is a potent competitor of Grb2 SH2-phosphopeptide interactions. This inhibitor has been obtained by rational drug design and is specific toward the Grb2 SH2 vs. other SH2 domains and the PTB domain of SHC in vitro. Accordingly, CGP78850 blocks epidermal growth factor receptor (EGFR)-Grb2 and Shc-Grb2 interactions in living cells. It also inhibits the growth of cells transformed by receptor tyrosine kinases, which transmit a proliferative signal through Grb2 to Ras, but not cells transformed by oncogenic Raf or cells that contain activating Ras mutations. Moreover, our results demonstrate that, in cells overexpressing receptor tyrosine kinases, such as the EGFR, Grb2 SH2 inhibitors induce expression of the cell cycle inhibitors p21(Waf1/Cip1/CAP1) and p27(Kip1) and reverse transformation. PMID- 10471534 TI - Modulation of urokinase-type plasminogen activator and metalloproteinase activities in cultured mouse mammary-carcinoma cells: enhancement by paclitaxel and inhibition by nocodazole. AB - Paclitaxel is a potent anti-tumor drug used in the treatment of breast cancer. It induces de-centralization of the microtubular system in tumor cells, blocking cell division. In the search for dissemination to a secondary site, cancer cells are capable of degrading most components of the extracellular matrix via an extracellular proteolytic cascade, including urokinase-type plasminogen activator (uPA) and the matrix metalloproteinases (MMPs). In the present study, the effects of paclitaxel and nocodazole, 2 drugs known to affect microtubules with opposite mechanisms of action, have been tested for their effect on the secretion of uPA and MMPs in cultures of F3II mouse mammary-tumor cells. Tumor-derived uPA activity significantly increased after pre-treatment of tumor cells for 24 hr with micromolar concentrations of paclitaxel (4 microM), while decreasing after pre-treatment with nocodazole (1 microM). A similar modulation was found for MMP 9 by zymographic analysis. Immunofluorescence and Western-blot analysis confirmed the formation of parallel microtubule fragments in paclitaxel-treated cells and almost complete de-polymerization of microtubules in nocodazole-treated ones. Our data suggest that, through opposite actions on microtubule organization and dynamics, paclitaxel and nocodazole exert inverse modulation of tumor-derived proteolytic activity in mammary tumor cells. PMID- 10471535 TI - TGF-beta-1 up-regulates cyclin D1 expression in COLO-357 cells, whereas suppression of cyclin D1 levels is associated with down-regulation of the type I TGF-beta receptor. AB - Transforming growth factor-beta1 (TGF-beta1) inhibits cell growth in susceptible cells by interacting with a family of protein kinases that control cell cycle progression. In the present study, we investigated the effects of TGF-beta1 on cyclin D1 expression and activity in COLO-357 human pancreatic cancer cells. TGF beta1 increased cyclin D1 mRNA and protein levels. Nuclear runoff transcription and protein synthesis inhibition by cycloheximide revealed that this increase was, in part, due to increased cyclin D1 mRNA synthesis. Despite its stimulatory effects on cyclin D1 levels, TGF-beta1 inhibited cyclin D1-associated kinase activity and the growth of COLO-357 cells. Furthermore, suppression of cyclin D1 expression with a cyclin D1 antisense cDNA resulted in loss of TGF-beta1-mediated growth inhibition in association with reduced induction of cyclin D1, p21(C)(ip)(1) and plasminogen activator inhibitor-1 (PAI-1). Concomitantly, there was a marked decrease in the levels of the type I TGF-beta receptor (TbetaRI). Our findings suggest that in some cell types cyclin D1 expression may be important for TGF-beta1-mediated signaling and that cyclin D1 may be involved in the transcriptional regulation of TbetaRI. PMID- 10471536 TI - Autocrine TGF-beta-regulated expression of adhesion receptors and integrin-linked kinase in HT-144 melanoma cells correlates with their metastatic phenotype. AB - We have previously shown that 2 human melanoma cell lines, the metastatic HT-144 and the non-metastatic SK-Mel-2 cells, exhibit marked in vitro heterogeneity with respect to integrin expression, migration and invasion potential. Here, we provide evidence that HT-144 melanoma cells, but not SK-Mel-2 cells, undergo a reversible transition to a fibroblastoid morphology following treatment with either their own serum-free acidified conditioned medium or biologically active exogenous TGF-beta1, thus identifying TGF-beta as an autocrine regulator of the spindle shape morphology of HT-144 melanoma cells. The fibroblastoid phenotype correlated with up-regulated beta1 and beta3 integrin and down-regulated E cadherin expression, as shown by flow cytometry, Western blot and RT-PCR, as well as up-regulated expression of the matrix metalloproteinase MMP-9, as demonstrated by zymography. Our data further illustrate the TGF-beta1-dependent up-regulation of integrin-linked kinase and the nuclear translocation of beta-catenin, 2 intracellular proteins involved in integrin and cadherin signaling. PMID- 10471537 TI - Additive antitumor activities of taxoids in combination with the bisphosphonate ibandronate against invasion and adhesion of human breast carcinoma cells to bone. AB - A very common metastatic site for breast carcinoma is bone. Metastatic breast carcinoma cells stimulate osteoclast-mediated bone resorption leading to osteolysis. Bisphosphonates are powerful inhibitors of osteoclast activity, and are therefore used in combination with standard chemotherapy or hormonal therapy for the treatment of cancer-associated osteolytic metastases. However, there may be an added beneficial effect of the bisphosphonates, that is, additive or synergistic activities with cytotoxic agents. Here, we investigated the effects of the bisphosphonate ibandronate in combination with taxoids (taxol and taxotere) on induction of apoptosis, invasion and adhesion of breast carcinoma cells to bone. Ibandronate did not induce apoptosis of human MDA-MB-231 breast carcinoma cells, nor did it enhance the effectiveness of taxoid-induced apoptosis in MDA-MB-231 cells. In contrast, ibandronate enhanced the antitumor activity of taxoids against invasion and cell adhesion to bone. Our findings raise the interesting possibility that the combination of bisphosphonates and taxoids may be useful for the treatment of patients with cancer types that are known to metastasize preferentially to bone. PMID- 10471538 TI - Bispecific antibody-mediated lysis of primary cultures of ovarian carcinoma cells using multiple target antigens. AB - We have shown previously that a bispecific antibody (BsAb) directed against both germ-cell alkaline phosphatase (GCAP) and the CD3 complex on mouse T cells could effectively eliminate GCAP-positive tumor cells in vivo using an immunocompetent mouse model. However, some GCAP-negative tumor cells were still able to grow, suggesting that BsAb therapy, when used in a clinical setting, could benefit from targeting several tumor markers to prevent outgrowth of tumor cells lacking a targeted marker. To test this hypothesis, we developed an in vitro model based on primary human ovarian carcinoma (OC) cultures and BsAbs directed against human T cells and several tumor markers [placental alkaline phosphatase (PLAP), GCAP, folate-binding protein (FBP) and CA19.9]. OC cells, isolated from primary tumors, were co-cultured with human peripheral blood mononuclear cells in the presence or absence of various concentrations of BsAbs against PLAP/GCAP, FBP and CA19.9 administered separately or in combination. Results derived from 18 primary OC samples showed that the combination treatment was better than or equally effective as the best single BsAB treatment in 60% of cases. Sometimes targeting FBP, PLAP/GCAP or CA19.9 alone was superior to targeting all simultaneously. Combining each BsAb with a low dose of IL-2 was always beneficial. These results indicate that before using a specific BsAb in the clinic, it is important to determine the optimal BsAb for each patient using this in vitro assay on cells from the removed tumor mass. PMID- 10471539 TI - Bone extracellular matrix stimulates invasiveness of estrogen-responsive human mammary MCF-7 cells. AB - Bone is the most frequent site of metastasis in breast cancer. This causes destructive osteolytic lesions. To achieve metastasis to bone, breast cancer cells must proliferate in a new microenvironment, arrest on extracellular matrix and invade. Breast cancer cells progress in the invasive processes only if they destroy bone with the assistance of osteoclasts. In this work, we present data suggesting that MCF-7 cells, an estradiol receptor-positive cell line that exhibits modest invasive capacity, proliferate in the presence of soluble factors secreted by the osteogenic cell line SaOS-2. The cells acquire a more aggressive phenotype when cultured on an extracellular matrix produced by the same osseous cell line. Acquisition of the invasive phenotype appears to be related to the capacity of bone extracellular matrix to induce the expression of urokinase-like plasminogen activator by MCF-7 cells, which is specific for MCF-7 cells, given that MDA-231 cells, an estradiol receptor-negative and more aggressive cell line, did not show significant changes when cultured in the presence of soluble and insoluble bone factors. PMID- 10471540 TI - Inhibition of human tumor cell growth in vitro and in vivo by a specific inhibitor of human farnesyltransferase: BIM-46068. AB - Oncogenic mutations of the ras gene leading to constitutive activation of downstream effectors have been detected in a large spectrum of human cancers (pancreas, thyroid, colon and NSCLC). Membrane anchorage of Ras required for functional activity in signal transduction is facilitated by post-translational modifications resulting in covalent attachment of a farnesyl group to the cysteine in the C-terminal CAAX motif. This attachment is mediated by farnesyltransferase (FTase). Here, we report a novel series of potent FTase inhibitors, where the tetrapeptide CAAX motif has been modified by incorporation of a thiazolidine carboxylic acid moiety followed by reduction of the 1st and 2nd peptide bonds to a secondary and tertiary amine, respectively. The C-terminal carboxylate was converted to esters for improved cellular penetration. These compounds showed specific inhibition of purified human FTase enzyme, inhibition of proliferation in vitro in a large spectrum of human tumor cell lines and inhibition of growth of human tumor xenografts in athymic nude mice. In addition, in regard to a panel of cell lines, using the Compare analysis to determine the Pearson coefficient correlation, the anti-proliferative spectrum of BIM-46068 has been shown to be distinct from the profile of typical chemotherapeutic agents. PMID- 10471541 TI - Synergism between vitamins E and C: biological implications for future research. PMID- 10471542 TI - Pharmacodynamic properties of St. John's wort-A single blind neurophysiological study in healthy subjects comparing two commercial preparations. AB - In this clinical trial the effects on the central nervous system (CNS) of two commercially available extracts of St. John's wort (Texx 300 and Jarsin 300) were compared with those of placebo in a group of healthy young volunteers (n = 35) following a single blind study design. Measurements were carried out on the first day and after repetitive drug application (t.i.d.) over 3 weeks. In accordance to the known slow resorption of hypericum extracts maximum effects were observed for both preparations between 4 and 6 h after intake. Texx 300 produced alpha2 increases of up to 32% in comparison to placebo. Somewhat weaker increases were observed after Texx 300 within the beta frequencies (14% resp. 15%) and in alpha1 band (8%). Jarsin 300 produced less power increases within the alpha2 range. In addition beta increases were observed after Jarsin 300 following repetitive administration. Thus slight differences could be detected between the two preparations with respect to their maximum effects, but in general the profile of action was similar for both drugs becoming more clear after the repetitive dosing. However, the effects of Texx 300 occurred a little bit earlier possibly due to a better intestinal resorption. Concerning the auditory event related potentials both preparations tended to decrease the latency of the cognitive potential P 300 indicating an improvement of mental performance. The observed neurophysiological changes are in line with the proposed clinical efficacy of both plant extracts, especially with respect to the drug induced changes within the alpha1 and alpha2 frequency ranges known for being under serotonergic and dopaminergic control, respectively. These electrochemical features of brain activity are known to be of great importance during the course of action of antidepressive drugs. - In summary quantitative EEG proved to be able to discriminate between different extracts of St. John's wort with respect to time of effect and profile changes on neuronal communication structure. PMID- 10471543 TI - Diagnostic value of chest radiography, computed tomography and tumour markers in the differentiation of malignant from benign solitary pulmonary lesions. AB - BACKGROUND: The aim of this prospective study was to assess the diagnostic value of the imaging modalities (chest radiography, spiral computed tomography (SCT) and high-resolution computed tomography (HRCT)) and the tumour markers (carcinoembryonic antigen (CEA), cytokeratin marker (CYFRA 21-1) and neuron specific enolase (NSE)) in the differentiation of malignant (MSPLs) from benign solitary pulmonary lesions (BSPLs). PATIENTS AND METHODS: Solitary pulmonary lesions (SPLs) were examined, evaluated and then completely removed by surgery in 104 consecutive patients (MSPLs n = 81, BSPLs n = 23). Chest radiography was performed with frontal and lateral views, SCT was carried out with a slice thickness of 8 mm and HRCT with a slice thickness of 1 mm and a 12-cm field of view. For the tumour marker analysis, serum concentrations were determined 1-3 days prior to surgery by ELISA for CEA and CYFRA 21-1 and by IRMA for NSE using commercially available assay kits. The cut-off values were set at 3 ng/ml (for non-smokers) and 5 ng/ml (for smokers) for CEA, at 3.3 ng/ml for CYFRA 21-1 and at 12.5 ng/ml for NSE. RESULTS: Using any one of the characteristics with a significance level of P <0.01, the identification of MSPLs using chest radiography showed a sensitivity of 64.2% and a specificity of 82.6%, using SCT a sensitivity of 88.9% and a specificity of 60.9% and using HRCT a sensitivity of 91.4% and a specificity of 56.5%. For the identification of MSPLs using CEA a sensitivity of 27.2% and a specificity of 87.0% (accuracy of 40.4%) was observed. Using CYFRA 21-1 a sensitivity of 19.8% and a specificity of 100.0% (accuracy of 37.5%) and using NSE a sensitivity of 13.6% and a specificity of 100. 0% (accuracy of 32.7%) was found. CONCLUSIONS: Using chest radiography, SCT and HRCT, a precise morphological assessment of the periphery of the pulmonary lesion and the adjacent visceral pleura is necessary to distinguish MSPLs from BSPLs. Tumour markers used alone or in combination with the imaging methods brought no additional benefits, in terms of sensitivity and accuracy, over the diagnostic imaging methods alone. However, the tumour markers exhibited a far superior specificity (100% for CYFRA 21-1 and NSE) compared with the imaging methods. PMID- 10471545 TI - Characteristics of flow dependency of nitric oxide in exhaled air in children with cystic fibrosis and asthma. AB - Nitric oxide (NO) is a free radical produced by the lungs which can easily be measured in exhaled air. NO may serve as a non-invasive marker for airway inflammation in chronic inflammatory diseases like asthma. However in patients with cystic fibrosis (CF) and severe airway involvemen normal or low levels of NO have been reported. To investigate this further we measured NO levels in exhaled air at 5 different flow rates in 14 asthmatics, 15 CF-patients and 13 healthy children. A dependency of exhaled NO on expiratory flow was demonstrated in all three groups. At slow flows lower NO levels in CF-patients and significantly higher levels in asthmatics compared to healthy individuals were found. When the data were fitted to a one compartment model of the lung described by NO(MOUTH) = NO(LUNG) - NO(LUNG) x e(-T/Vex) (T = transfer factor; Vex = expiratory flow), NO(LUNG) was increased in asthmatics (191.9 +/- 53.8 ppb) and low in CF (26.7 +/- 5.7 ppb) compared to healthy individuals (76.9 +/- 50.9 ppb; p(anova) = 0.0213). NO produced in the central compartment of the lung behaved similarly and was distinguished from a peripheral compartment with the two compartment model NO(MOUTH) = NO(central) - (NO(central) - NO(peripher) ) x e(-T/Vex). We conclude that NO in exhaled air is flow dependent and at slow expiratory flows elevated in asthmatics and reduced in CF-patients compared to healthy children. Concentrations extrapolated for the whole lung and for the central airways changed proportionally. PMID- 10471544 TI - Increased levels of vascular endothelial growth factor in bronchoalveolar lavage of patients with bronchial carcinoma effect of tumour activity and oxidative stress due to radio-chemotherapy? AB - BACKGROUND: Vascular endothelial growth factor (VEGF) plays a crucial role in physiological and neoplastic angiogenesis. Moreover, VEGF has been found to be upregulated by conditions associated with the generation of free radicals and reactive oxygen intermediates. In patients with cancer, studies to evaluate VEGF as a measure of tumour activity were carried out. We tested the hypothesis that VEGF is additionally affected by oxidative stress due to anticancer therapy. Moreover, the suitability of epidermal growth factor (EGF) to estimate tumour activity was studied. PATIENTS AND METHODS: 60 patients with non-small cell lung cancer (NSCLC) covering different therapy progress and modalities underwent bronchoalveolar lavage. VEGF-, EGF-, albumin- and total protein-concentrations in bronchoalveolar lavage fluid (BALF) and VEGF-levels in blood plasma were studied. RESULTS: BALF VEGF-levels were increased in patients with advanced NSCLC before and in anticancer therapy. In patients who had received radiotherapy to the lung prior to chemotherapy, VEGF concentrations were noticeably higher than under sole chemotherapy. Pulmonary endothelial hyperpermeability was found in patients with recently diagnosed tumours and patients undergoing anti-cancer therapy. Evaluation of EGF-levels in BALF revealed no significant influence of tumour activity or cancer therapy on this parameter. CONCLUSION: BALF-levels of VEGF are affected by tumour activity and oxidative stress due to anticancer therapy. PMID- 10471546 TI - HIV and AIDS in Africa: impact on mother and child health. AB - Mother to child transmission of HIV-1 is a leading challenge of public health in developing countries. Various risk factors for vertical transmission of HIV have been identified, and both prenatal and postnatal infection by breast feeding occur. A bundle of interventions has lead to a dramatic drop of infection rates in industrialised countries. For the developing countries it is crucial to increase accessibility of health services, to perform prevention campaigns, and to provide antiretroviral prophylaxis for pregnant women. PMID- 10471547 TI - Liquid pancreatic enzyme therapy for a patient with short bowel syndrome and chronic pancreatitis in a complicated case of Crohn's disease. AB - The case of a 45 year old female with multiple complications of Crohn's disease is reported. After multiple resections in the gastrointestinal tract she had been suffering from short bowel syndrome and severe malnutrition. With a special continuous gastric tube feeding system, she was able to maintain her weight for years. In the beginning of 1997 the enteral nutrition was not longer tolerated for an exacerbation of chronic pancreatitis. There was a weight loss, permanent pain and total parenteral nutrition had to be performed. In this situation a new liquid preparation of pancreatic enzymes which had been tested in the laboratory before, was used for continuous enzyme replacement via gastric tube. In combination with this enzyme preparation, enteral nutrition could successfully be started again. PMID- 10471548 TI - Antibiotic resistance in Listeria spp. PMID- 10471549 TI - Activity of pleconaril against enteroviruses. AB - The activity of pleconaril in cell culture against prototypic enterovirus strains and 215 clinical isolates of the most commonly isolated enterovirus serotypes was examined. The latter viruses were isolated by the Centers for Disease Control and Prevention during the 1970s and 1980s from clinically ill subjects. Pleconaril at a concentration of 16; for Bacteroides ovatus (12), 4 and >16; for Bacteroides stercoris (12), 0.5 and 0.5; for Bacteroides uniformis (12), 1 and 4; for Bacteroides vulgatus (11), 4 and 4; for Clostridium clostridioforme (15), 0.5 and 0.5; for Clostridium difficile (15), 1 and >16; for Clostridium innocuum (13), 0.125 and 2; for Clostridium perfringens (13), 0.06 and 0.06; for Clostridium ramosum (14), 0.25 and 8; for Fusobacterium nucleatum (12), 0.125 and 0.25; for Fusobacterium necrophorum (11), 0.25 and 0.5; for Fusobacterium varium (13), 0.5 and 1; for Fusobacterium spp. (12), 1 and 2; for Peptostreptococcus anaerobius (13), 0.06 and 0.06; for Peptostreptococcus asaccharolyticus (13), 0.125 and 0.125; for Peptostreptococcus magnus (14), 0.03 and 0.03; for Peptostreptococcus micros (12), 0.06 and 0.06; for Peptostreptococcus prevotii (14), 0.06 and 0.25; for Porphyromonas asaccharolytica (11), 0.125 and 0.125; for Prevotella bivia (10), 8 and 16; for Prevotella buccae (10), 2 and 2; for Prevotella intermedia (10), 0.5 and 0.5; and for Prevotella melaninogenica (11), 1 and 1. Gemifloxacin mesylate (SB 265805) was 1 to 4 dilutions more active than trovafloxacin against fusobacteria and peptostreptococci, and the two drugs were equivalent against clostridia and P. asaccharolytica. Gemifloxacin was equivalent to sitafloxacin (DU 6859a) against peptostreptococci, C. perfringens, and C. ramosum, and sitafloxacin was 2 to 3 dilutions more active against fusobacteria. Sparfloxacin, grepafloxacin, and levofloxacin were generally less active than gemifloxacin against all anaerobes. PMID- 10471571 TI - In vitro activities of BMS-207147 against over 600 contemporary clinical bloodstream isolates of Candida species from the SENTRY Antimicrobial Surveillance Program in North America and Latin America. AB - We compared the in vitro activity of BMS-207147, an investigational triazole, with those of itraconazole and fluconazole against 613 clinical bloodstream isolates of Candida spp. collected from SENTRY participating hospitals during 1997 and 1998. Overall, BMS-207147 was the most active azole against all Candida spp. While both BMS-207147 and itraconazole displayed a stepwise decrease in activity against isolates for which the fluconazole MICs were elevated, BMS 207147 had two- to fourfold greater activity than itraconazole both against Candida spp. that were dose-dependently fluconazole susceptible and against those that were fluconazole resistant. PMID- 10471572 TI - Synergistic effect of clindamycin and atovaquone in acute murine toxoplasmosis. AB - The effect of clindamycin (CLI) combined with autovaquone (ATO) was examined in a murine model of acute toxoplasmosis. Swiss Webster mice intraperitoneally infected with 10(2) or 10(4) tachyzoites of the RH strain of Toxoplasma gondii were perorally treated with either drug alone (for ATO, 5, 25, 50, or 100 mg/kg of body weight/day; for CLI, 25, 50, or 400 mg/kg/day) or both combined (for ATO plus CLI, respectively, 5 plus 25, 25 plus 25, 25 plus 50, 50 plus 50, or 100 plus 400 mg/kg/day) starting with day 1 for 14 days. Survival was monitored during 7 weeks. Residual infection was assessed by a bioassay of representative 4 week survivors and by parasite DNA detection by PCR for representative 7-week survivors. An effect of treatment was shown in all treatment groups compared to untreated control mice (P = 0.0000). Among mice infected with 10(2) parasites, ATO and CLI at any dose combination protected significantly more animals than ATO alone (P = 0.0000), but compared to CLI alone, given its good effect, the combined drugs were no more effective (P > 0.05). For mice infected with 10(4) parasites, the drugs combined at the lowest and highest doses (5 plus 25 and 100 plus 400 mg/kg/day) were, similarly, more effective than ATO alone (P = 0.035 and 0.000, respectively) but not than CLI alone (P > 0. 05). However, treatment with ATO plus CLI at 25 plus 25, 25 plus 50, and 50 plus 50 mg/kg/day protected 20, 33, and 78% of mice, respectively, compared to virtually no survivals among those treated with either drug alone (P < 0.0005), thus demonstrating a significant synergistic effect of ATO and CLI against T. gondii. Furthermore, the dose of ATO at a given dose of CLI was shown to be critical to the effect. Moreover, the absence of residual infection in some survivors shows the potential of this drug combination to eliminate the parasite. PMID- 10471573 TI - Quantitation of intracellular triphosphate of emtricitabine in peripheral blood mononuclear cells from human immunodeficiency virus-infected patients. AB - An analytical methodology combining solid-phase extraction (SPE) and high performance liquid chromatography (HPLC) was developed to quantitate the intracellular active 5'-triphosphate (TP) of beta-L-2',3'-dideoxy-5-fluoro-3' thiacytidine (emtricitabine) (FTC) in human peripheral blood mononuclear cells (PBMCs). The FTC nucleotides, including 5'-mono-, di-, and triphosphates, were successively resolved on an anion-exchange SPE cartridge by applying a gradient of potassium chloride. The FTC-TP was subsequently digested to release the parent nucleoside that was finally analyzed by HPLC with UV detection (HPLC-UV). Validation of the methodology was performed by using PBMCs from healthy donors exposed to an isotopic solution of [(3)H]FTC with known specific activity, leading to the formation of intracellular FTC-TP that was quantitated by an anion exchange HPLC method with radioactive detection. These levels of FTC-TP served as reference values and were used to validate the data obtained by HPLC-UV. The assay had a limit of quantitation of 4. 0 pmol of FTC-TP (amount on column from approximately 10(7) cells). Intra-assay precision (coefficient of variation percentage of repeated measurement) and accuracy (percentage deviation of the nominal reference value), estimated by using quality control samples at 16.2, 60.7, and 121.5 pmol, ranged from 1.3 to 3.3% and -1.0 to 4. 8%, respectively. Interassay precision and accuracy varied from 3.0 to 10.2% and from 2.5 to 6.7%, respectively. This methodology was successfully applied to the determination of FTC-TP in PBMCs of patients infected with human immunodeficiency virus after oral administration of various dosing regimens of FTC monotherapy. PMID- 10471574 TI - Fluoroquinolone resistance in Bacteroides fragilis following sparfloxacin exposure. AB - In vitro pharmacodynamic studies investigating the antimicrobial properties of five fluoroquinolones, (trovafloxacin, sparfloxacin, clinafloxacin, levofloxacin, and ciprofloxacin) against Bacteroides fragilis ATCC 23745 were conducted. The times required to reduce the viable counts by 3 log units were as follows: clinafloxacin, 2.9 h; levofloxacin, 4.6 h; trovafloxacin, 6 h; and sparfloxacin, 10 h. Exposure to ciprofloxacin did not achieve a 3-log decrease in viable counts. The susceptibility of B. fragilis was determined both prior to exposure and following 24 h of exposure to each of the five fluoroquinolones tested. The MICs of clinafloxacin, levofloxacin, trovafloxacin, sparfloxacin, ciprofloxacin, metronidazole, cefoxitin, chloramphenicol, and clindamycin were determined by the broth microdilution method. The MICs for B. fragilis preexposure were as follows: clinafloxacin, 0.25 microg/ml; trovafloxacin, 0.5 microg/ml; sparfloxacin, 2 microg /ml; levofloxacin, 2 microg/ml; and ciprofloxacin, 8 microg/ml. Similar pre- and postexposure MICs were obtained for cultures exposed to trovafloxacin, clinafloxacin, levofloxacin, and ciprofloxacin. However, following 24 h of exposure to sparfloxacin, a fluoroquinolone-resistant strain emerged. The MICs for this strain were as follows: clinafloxacin, 1 microg/ml; trovafloxacin, 4 microg/ml; sparfloxacin, 16 microg/ml; levofloxacin, 16 microg/ml; and ciprofloxacin, 32 microg/ml. No changes in the susceptibility of B. fragilis pre- and postexposure to sparfloxacin were noted for metronidazole (MIC, 1 microg/ml), cefoxitin (MIC, 4 microg /ml), chloramphenicol (MIC, 4 microg/ml), and clindamycin (MIC, 0.06 microg/ml). Resistance remained stable as the organism was passaged on antibiotic-free agar for 10 consecutive days. Mutant B. fragilis strains with decreased susceptibility to clinafloxacin, trovafloxacin, sparfloxacin, levofloxacin, and ciprofloxacin were selected on brucella blood agar containing 8x the MIC of levofloxacin at a frequencies of 6.4 x 10(-9), 4x the MICs of trovafloxacin and sparfloxacin at frequencies of 2.2 x 10(-9) and 3. 3 x 10(-10), respectively, and 2x the MIC of clinafloxacin at a frequency of 5.5 x 10(-11); no mutants were selected with ciprofloxacin. The susceptibilities of strains to trovafloxacin, levofloxacin, clinafloxacin, sparfloxacin, and ciprofloxacin before and after exposure to sparfloxacin were modestly affected by the presence of reserpine (20 microg/ml), an inhibitor of antibiotic efflux. The mechanism of fluoroquinolone resistance is being explored, but it is unlikely to be efflux due to a lack of cross-resistance to unrelated antimicrobial agents and to the fact that the MICs for strains before and after exposure to sparfloxacin are minimally affected by reserpine. PMID- 10471575 TI - Histatin 3-mediated killing of Candida albicans: effect of extracellular salt concentration on binding and internalization. AB - Human saliva contains histidine-rich proteins, histatins, which have antifungal activity in vitro. The mechanism by which histatins are able to kill Candida albicans may have clinical significance but is currently unknown. Using radiolabeled histatin 3, we show that the protein binds to C. albicans spheroplasts in a manner that is dependent on time and concentration. Binding to the spheroplasts was saturable and could be competed with unlabeled histatin 3. A single histatin 3 binding site with a K(d) = 5.1 microM was detected. Histatin 3 binding resulted in potassium and magnesium efflux, predominantly within the first 30 min of incubation. Studies with fluorescent histatin 3 demonstrate that the protein is internalized by C. albicans and that translocation of histatin inside the cell is closely associated with cell death. Histatin binding, internalization, and cell death are accelerated in low-ionic-strength conditions. Indeed, a low extracellular salt concentration was essential for cell death to occur, even when histatin 3 was already bound to the cell. The interaction of histatin 3 with C. albicans, and subsequent cell death, is inhibited at low temperature. These results demonstrate that the candidacidal activity of histatin 3 is not due exclusively to binding at the cell surface but also involves subsequent interactions with the cell. PMID- 10471576 TI - Oxfendazole treatment of sheep with naturally acquired hydatid disease. AB - A blinded, randomized placebo-controlled trial assessed the efficacy and safety of oxfendazole for the treatment of ovine hydatid disease. Cyst fertility and parasite viability were measured following daily, weekly, and monthly treatment schedules with 30 mg of oxfendazole per kg of body weight. The 12-week trial was conducted in 215 adult sheep in the central Peruvian Andes and was masked for both treatment group and scheduling. In this trial oxfendazole significantly reduced protoscolex viability relative to controls in all treatment groups. In the daily, weekly, and monthly groups, 100, 97, and 78% of sheep, respectively, were either cured or improved following treatment, compared to 35% cured or improved animals in the control group. However, daily dosing at 30 mg of oxfendazole per kg proved highly toxic to sheep, resulting in a 24% death rate in the daily group as compared to a 4 to 6% mortality rate in all other groups. If found safe in humans, oxfendazole may prove to be a useful and inexpensive treatment for cestode infections in humans. This study suggests that a staggered dosing regimen of oxfendazole, and possibly other benzimidazoles, may be as efficacious as daily treatment regimens for hydatidosis while decreasing both the cost and adverse effects associated with daily dosing. PMID- 10471577 TI - In vitro activities of azithromycin and ofloxacin against Chlamydia pneumoniae in a continuous-infection model. AB - Chlamydia pneumoniae is a well-established cause of community-acquired pneumonia and bronchitis in adults and children. Chronic infections with C. pneumoniae have been implicated in the development of atherosclerosis and other diseases in humans. Methods currently used for the culture and propagation of C. pneumoniae are not analogous to the infection as it occurs in vivo. We have established a model of continuous C. pneumoniae infection in vitro. HEp-2 cells inoculated with CM-1 and TW-183 strains have been persistently infected for periods of over 1.5 and 2 years, respectively. The cultures were maintained without centrifugation or the addition of cycloheximide, fresh host cells, or chlamydia. We observed cycles of host cell lysis, detachment, and regrowth with both strains of C. pneumoniae. Continuous C. pneumoniae infections may more closely resemble the actual events as they occur in vivo and, therefore, may be a better model for the in vitro study of C. pneumoniae infection. When we used continuously infected cells to determine the effects of azithromycin and ofloxacin on C. pneumoniae propagation in vitro, we found that both drugs reduced but did not completely eliminate the organism. This may be an important observation, as the failure of antibiotic therapy against C. pneumoniae infection in humans has been described. PMID- 10471578 TI - Inhibition of Escherichia coli-induced meningitis by carboxyfullerence. AB - The effect of a water-soluble malonic acid derivative of carboxyfullerence (C60) against Escherichia coli-induced meningitis was tested. C60 can protect the mice from E. coli-induced death in a dose-dependent manner. C60 administered intraperitoneally as late as 9 h after E. coli injection was still protective. The C60-treated mice had less tumor necrosis factor alpha and interleukin-1beta production by staining of brain tissue compared to the levels of production for nontreated mice. The E. coli-induced increases in blood-brain barrier permeability and inflammatory neutrophilic infiltration were also inhibited. These data suggest that C60 is a potentially therapeutic agent for bacterial meningitis. PMID- 10471579 TI - Incidence of quinolone resistance over the period 1986 to 1998 in veterinary Salmonella isolates from Germany. AB - A total of 24,591 nonhuman salmonella strains isolated in Germany between 1986 and 1998 were examined for their resistance to nalidixic acid by an agar diffusion method. The rate of resistance (inhibition zone, /=1 microg/ml. PMID- 10471583 TI - In vitro activities of fluoroquinolones against antibiotic-resistant blood culture isolates of viridans group streptococci from across Canada. AB - Among 418 blood culture isolates of viridans group streptococci obtained between 1995 and 1997, the in vitro rates of nonsusceptibility to penicillin, erythromycin, tetracycline, and trimethoprim-sulfamethoxazole were 28, 29, 24, and 14%, respectively. The most prevalent group (125 strains) was Streptococcus mitis, followed by Streptococcus sanguis (56 strains). For 236 (56%) strains resistant to one or more antibiotics, the ciprofloxacin MIC at which 90% of the isolates were inhibited (MIC(90)) was 4 microg/ml, whereas the MIC(90)s of trovafloxacin, grepafloxacin, and gatifloxacin were 0.25 microg/ml. PMID- 10471584 TI - Mononuclear and polymorphonuclear leukocyte dispositions of clarithromycin and azithromycin in AIDS patients requiring Mycobacterium avium complex prophylaxis. AB - The intracellular dispositions of clarithromycin and azithromycin in AIDS patients requiring Mycobacterium avium complex (MAC) prophylaxis were studied. The dispositions of both drugs in mononuclear and polymorphonuclear leukocytes were markedly different. Our data support the proven efficacy of these agents for MAC prophylaxis since clarithromycin and azithromycin displayed sustained intracellular concentrations which exceeded their MICs for MAC throughout the dosing periods. PMID- 10471585 TI - Lack of in vitro antimicrosporidian activity of thalidomide. AB - Thalidomide was evaluated for its in vitro activity against Encephalitozoon species by using the MRC-5 cell system. A cytotoxic effect was observed for concentrations of 10(1) microg/ml (P < 10(5)) and 5 microg/ml (P < 10(5)). Thalidomide did not significantly inhibit the growth of any of the microsporidia under study. These data suggest that thalidomide is not an etiologic treatment in microsporidial enteritis. PMID- 10471586 TI - Cefoperazone prevents the inactivation of alpha(1)-antitrypsin by activated neutrophils. AB - At sites of neutrophilic inflammation, tissue injury by neutrophil elastase is favored by phagocyte-induced hypochlorous acid-dependent inactivation of the natural elastase inhibitor alpha(1)-antitrypsin. In the present study, cefoperazone prevented alpha(1)-antitrypsin inactivation by neutrophils and reduced the recovery of hypochlorous acid from these cells. Moreover, the antibiotic reduced the free elastase activity in a neutrophil suspension supplemented with alpha(1)-antitrypsin without affecting the cells' ability to release elastase. These data suggest that the drug inactivates hypochlorous acid before its reaction with alpha(1)-antitrypsin, thereby permitting the antiprotease-mediated blockade of released elastase. In conclusion, cefoperazone appears to have the potential for limiting elastase-antielastase imbalances, attenuating the related tissue injury at sites of inflammation. PMID- 10471587 TI - Antimicrobial susceptibility testing of Chlamydia trachomatis using a reverse transcriptase PCR-based method. AB - The conventional method for antimicrobial susceptibility testing of Chlamydia trachomatis is subjective and potentially misleading. We have developed a reverse transcriptase PCR (RT-PCR)-based method which is more sensitive and less subjective than the conventional method. Using 16 strains of C. trachomatis in triplicate assays, we found the RT-PCR method consistently more sensitive than the conventional technique for all eight antimicrobials tested, with resultant MICs determined by RT-PCR ranging from 1.6-fold higher (erythromycin) to >/=195 fold higher (amoxicillin). PMID- 10471588 TI - Bactericidal/permeability-increasing protein inhibits growth of a strain of Acholeplasma laidlawii and L forms of the gram-positive bacteria Staphylococcus aureus and Streptococcus pyogenes. AB - Bactericidal/permeability-increasing protein (BPI) inhibited growth of cell wall deficient Acholeplasma laidlawii and L forms of certain strains of Staphylococcus aureus and Streptococcus pyogenes. However, the same strains of S. aureus and S. pyogenes with intact cell walls were not susceptible to the growth-inhibitory effects of BPI. PMID- 10471589 TI - Relationship between pyrazinamide resistance, loss of pyrazinamidase activity, and mutations in the pncA locus in multidrug-resistant clinical isolates of Mycobacterium tuberculosis. AB - Sixty-two Mycobacterium tuberculosis isolates were tested for pyrazinamidase activity, and their pyrazinamide susceptibility was determined by the radiometric method. Sequencing of pncA genes in the 23 resistant strains revealed mutations in 16 pyrazinamidase-negative strains, 11 of which had not been previously described. Six isolates containing wild-type pncA might possess alternative resistance mechanisms. PMID- 10471590 TI - Activities of new antimicrobial agents (trovafloxacin, moxifloxacin, sanfetrinem, and quinupristin-dalfopristin) against Bacteroides fragilis group: comparison with the activities of 14 other agents. AB - The antimicrobial activities of trovafloxacin, moxifloxacin, sanfetrinem, quinupristin-dalfopristin, and 14 other antimicrobial agents against 218 Bacteroides fragilis group strains were determined. A group of 10 imipenem resistant strains were also tested. Imipenem, meropenem, and sanfetrinem had the lowest MICs of all of the beta-lactams. Quinupristin-dalfopristin inhibited all of the strains at 2 microg/ml. Overall, the MICs of trovafloxacin and moxifloxacin for 90% of the strains tested were 1 and 2 microg/ml, respectively. PMID- 10471591 TI - Pharmacokinetics and safety of high-dose and extended-interval regimens of levofloxacin in human immunodeficiency virus-infected patients. AB - The pharmacokinetics of levofloxacin, administered in high doses and with extended dosing intervals, was studied in human immunodeficiency virus (HIV) infected patients. Thirty patients received either 750 mg of the drug or a placebo once daily for 14 days, followed by 750 mg or 1,000 mg of the drug or a placebo three times weekly for an additional 14 days. Levofloxacin disposition was characterized by rapid oral absorption, with peak concentrations occurring approximately 1.5 h after dosing and elimination half-lives from 7.2 to 9.4 h. The overall incidence of any adverse effect was 70% (1,000 mg) to 95% (750 mg) for levofloxacin-treated patients and 71% for those taking the placebo. Levofloxacin pharmacokinetic parameters for HIV-infected patients were consistent with those observed in studies of healthy volunteers. PMID- 10471593 TI - The torch passes, the AJRCCM community stays aglow PMID- 10471592 TI - In vitro activities of strychnos alkaloids and extracts against Plasmodium falciparum. AB - The in vitro antimalarial activities of 46 alkaloids and extracts from Strychnos species were evaluated. Two types of quasidimeric alkaloids exhibit high and selective activities against Plasmodium. Strychnopentamine and isostrychnopentamine were active against chloroquine-sensitive and -resistant strains (50% inhibitory concentration [IC(50)] approximately 0.15 microM), while dihydrousambarensine exhibited a 30-fold higher activity against the chloroquine resistant strain (IC(50) = 0.03 microM) than it did against the chloroquine sensitive strain. PMID- 10471594 TI - Therapeutic bronchoscopy in broncholithiasis. AB - Bronchoscopy is considered the most important diagnostic test for broncholithiasis. However, its role in the treatment of broncholithiasis in a large group of patients has not been studied. To evaluate the therapeutic role of bronchoscopy, we retrospectively reviewed the clinical data of patients with broncholithiasis who also underwent bronchoscopy at Mayo Clinic. Bronchoscopy revealed 127 broncholiths (free or partly eroded calcified material in the airway lumen) in 95 patients (49 men and 46 women) evaluated between 1954 and 1994. Bronchoscopic removal of 71 (56%) broncholiths was attempted in 48 patients (50.5%) during 61 bronchoscopy sessions. Forty-eight of the broncholiths selected for removal were partly eroding into the tracheobronchial lumen and 23 were free. Forty-eight percent (23 of 48) of the partly eroding broncholiths were successfully removed bronchoscopically, with a greater percentage removed with the rigid bronchoscope (67%) than with the flexible bronchoscope (30%). All free broncholiths were completely extracted regardless of the type of bronchoscope used. Complications occurred in only two patients (4% of the bronchoscopic broncholithectomy group), both with partially eroded broncholiths, and consisted of hemorrhage in one patient requiring thoracotomy and acute dyspnea in another patient, caused by a loose broncholith lodged in the trachea. We conclude that flexible and/or rigid bronchoscopic extraction of partly eroded or free broncholiths in the tracheobronchial tree can be considered safe and effective. PMID- 10471595 TI - Comparison of two methods of diagnostic lung lavage in ventilated infants with lung disease. AB - The methods of nonbronchoscopic lung lavage used for collection of samples of epithelial lining fluid (ELF) in intubated patients are poorly standardized and incompletely validated. In infants with lung disease requiring ventilatory support, we evaluated two techniques of small volume saline lavage for the collection of a specimen suitable for pulmonary surfactant analysis. We aimed to compare apparent origin of the return fluid obtained by each method, equivalence and agreement of the estimates of measured pulmonary surfactant concentration, and the relative strength of association between surfactant indices and lung dysfunction. Fifty-three contemporaneous paired samples of lung lavage fluid suitable for surfactant analysis were collected from 31 infants using tracheal aspirate (TA, 4 x 0.5 ml saline), and then nonbronchoscopic bronchoalveolar lavage (NB-BAL, 3 x 1 ml/kg). Return fluid from TA had higher mean ELF concentration of total protein and IgA secretory component (SC), and a lower surfactant protein A (SP-A) concentration than NB-BAL, indicating that the TA lavage was sampling ELF more proximally in the tracheobronchial tree (protein: TA 7.7 versus NB-BAL 4.7 mg/ml; SC: 21 versus 1.8 microgram/ml; SP-A: 9.8 versus 19 microgram/ml; all p < 0.01). Mean concentration of surfactant indices in ELF differed only for SP-A, but for all indices, paired values showed poor agreement on Bland-Altman analysis, highlighting the potential imprecision associated with small volume lung lavage. TA return fluid yielded estimates of surfactant indices which were at least equivalent to NB-BAL in prediction of the severity of lung dysfunction. We conclude that NB-BAL return fluid has more distal origin, but analysis of TA fluid may have equal validity in the estimation of indices of pulmonary surfactant. The results of individual estimates of ELF constituents in a single sample of lavage fluid should be interpreted with caution, even when standardized sampling techniques are employed. PMID- 10471596 TI - Validity of markers of dilution in small volume lung lavage. AB - Definitive analysis of solute concentrations in lung lavage fluid involves the use of a marker of dilution to correct for variable recovery of epithelial lining fluid (ELF), but the question of the most appropriate dilutional marker remains unresolved. In lavage fluid collected from infants with lung disease and healthy control subjects, we examined ELF concentration of protein, albumin, sphingomyelin (SM), and IgA secretory component (SC), and critically appraised the relative validity of SC and urea as dilutional markers in the context of lung infection and lung injury. Protein, albumin, and SM were found not to be valid dilutional markers, as their ELF concentration varied significantly between the diseased, recovering, and normal lung. Differences in concentration were noted in both tracheal aspirate samples (TA, 4 x 0.5 ml) and nonbronchoscopic bronchoalveolar lavage fluid (NB-BAL, 3 x 1 ml/kg), but were not uniform (e.g., TA-disease versus control: albumin 2.8 versus 0.68 mg/ml, SM 45 versus 16 microgram/ml, both p < 0.05; NB-BAL-disease versus recovery: protein 8.1 versus 4.8 mg/ml, albumin 2.9 versus 1. 4 mg/ml, both p < 0.05). Overall, SC concentrations in ELF were not different between the diseased and normal lung, but in the NB-BAL samples, significantly higher SC concentration was noted in viral bronchiolitis and pneumonia than in noninfective lung diseases. No clear evidence of additional influx of urea into lavage fluid in association with epithelial disruption was found in the diseased lung. Comparative analysis of SC and urea revealed no difference in TA samples, but in NB-BAL specimens, urea best standardized the lavage concentration of surfactant indices to correspond to the degree of lung dysfunction as indicated by oxygenation index. We conclude that SC and urea, but not protein, albumin, or SM, are valid dilutional markers with which to estimate ELF recovery during small volume lung lavage. Urea appears a more appropriate choice in return fluid derived from the distal tracheobronchial tree, and SC should not be used in the context of lung infection. PMID- 10471597 TI - Correcting static intrinsic positive end-expiratory pressure for expiratory muscle contraction. Validation of a new method. AB - We have recently shown (Eur. Respir. J. 1997;10:522-529) that in spontaneously breathing and actively expiring patients, static intrinsic positive end expiratory pressure (PEEPi,st) can be corrected for expiratory muscle contraction by subtracting the average expiratory rise in gastric pressure (Pga,exp rise), calculated from three breaths just prior to an airway occlusion, from the end expiratory airway pressure (Paw) of the first occluded inspiratory effort (PEEPi,st avg). However, since in some patients there is substantial variability in the intensity of expiratory muscle activity and hence in Pga,exp rise, this method may be inaccurate because the Pga,exp rise of breaths preceding airway occlusion may differ from that of the first postocclusion breath. In the present study, we introduced a new method consisting of synchronous subtraction of Pga,exp rise from Paw, both occurring during airway occlusion (PEEPi,st sub). PEEPi,st sub and PEEPi,st avg were each compared with the reference PEEPi,st (PEEPi,st ref), which was obtained during muscular paralysis and simulation of the spontaneous breathing pattern by the ventilator. We found that, in 25 critically ill patients, PEEPi,st sub (mean +/- SD, 5.3 +/- 2.6 cm H(2)O) was nearly identical to PEEPi,st ref (5.4 +/- 2.4 cm H(2)O). Their mean difference was -0.06 cm H(2)O with limits of agreement -0.96 to 0.84 cm H(2)O, indicating a strong agreement between these methods. In contrast, mean difference of PEEPi,st avg and PEEPi,st ref was 0.73 cm H(2)O with limits of agreement -3.97 to 5.43 cm H(2)O, indicating lack of agreement. Coefficient of variation of Pga,exp rise was 14.3 +/- 7.2% (range, 5.2 to 28.3%). There was a good correlation between the coefficient of variation of Pga,exp rise and the difference between PEEPi,st avg and PEEPi,st ref (r = 0.909; p < 0.001). We conclude that PEEPi,st can be accurately measured in spontaneously breathing patients by synchronous subtraction of Pga,exp rise from Paw during airway occlusion. PMID- 10471598 TI - Respiratory syncytial and other virus infections in persons with chronic cardiopulmonary disease. AB - Respiratory syncytial virus (RSV) has been increasingly recognized as an important cause of serious respiratory illness in some adult populations, including those with underlying cardiopulmonary diseases. However, the precise incidence and the clinical impact of RSV in this group are unknown. Therefore, the incidence and clinical impact of RSV infection in persons with chronic obstructive pulmonary disease (COPD) and congestive heart failure (CHF) who reside in the community were prospectively evaluated over two consecutive winters in 134 persons. Eight RSV (incidence of 4.3 per 100 subject-winters), 13 influenza A (incidence of 7.0 per 100 subject-winters), seven rhinovirus, nine coronavirus, and two parainfluenza virus infections were identified. The clinical illnesses associated with RSV and influenza A virus were similar, causing both upper and lower respiratory signs and symptoms. The clinical impact was significant as three of eight RSV-infected subjects were hospitalized compared with six of 13 influenza A-infected persons and zero of seven rhinovirus-infected persons. PMID- 10471599 TI - Increased concentrations of iron and isoferritins in the lower respiratory tract of patients with stable cystic fibrosis. AB - Reactive oxygen species may contribute to airway injury in patients with cystic fibrosis (CF) and iron catalyzes oxidant injury by promoting generation of highly reactive hydroxyl radicals. Iron in the lower respiratory tract may be free, ferritin bound (from which iron can be reductively mobilized), or transferrin bound (which generally prevents iron mobilization). Ferritin is composed of subunits that are heavy (H) or light (L), and H-rich ferritins have additional biologic effects including inhibition of lymphocyte proliferation and cell growth. To assess concentrations of iron and iron-binding proteins in the lower respiratory tract of patients with CF, we measured iron (ferrozine), L-ferritin, H-ferritin, and transferrin (enzyme-linked immunosorbent assay [ELISA]) in bronchoalveolar lavage (BAL) fluid recovered from stable patients with CF (n = 8), healthy nonsmokers (NS; n = 8), or heavy cigarette smokers (HS; n = 8). Iron was detected in BAL fluid from patients with CF and HS, but not NS, with higher iron concentrations in patients with CF (42.0 +/- 11.6 microgram/dl) than in HS (9.9 +/- 2.6 microgram/dl, p < 0.05). Ferritin was present in all BAL fluids, with higher total ferritin (L + H) in patients with CF (647 +/- 84 ng/ml) than in HS (181 +/- 25 ng/ml, p < 0.005) or NS (9 +/- 3 ng/ml, p < 0.0005). Ferritin recovered from HS and NS lungs was < 2% H type, whereas ferritin in CF lungs was > 40% H-type ferritin. Transferrin concentrations in BAL fluid were not different in any group. Tumor necrosis factor (TNF)-alpha was present only in BAL samples from patients with CF. To assess whether TNF-alpha contributed to H-ferritin accumulation in CF lungs, we treated lung epithelial cells (A549) with iron alone (FeSO(4), 10-40 microM) or with iron and TNF-alpha (5-20 ng/ml). Iron-treated A549 cells synthesized almost entirely L-ferritin whereas exposure to TNF-alpha with iron caused a dose-dependent increase in accumulation of H-type ferritin. These findings suggest that oxidant injury could be promoted in lungs of patients with cystic fibrosis by iron mobilized from extracellular ferritin and, in addition, that TNF-alpha-promoted accumulation of H-type ferritin may impair local immune function and cell growth. PMID- 10471600 TI - Modulation of elastase binding to elastin by human alveolar macrophage-derived lipids. AB - Human neutrophil elastase (HNE), an enzyme secreted by activated neutrophils, can bind to and degrade extracellular matrix including human lung elastin. This protease is believed to play an important role in several destructive processes including pulmonary emphysema. In this study, we hypothesized that an alveolar macrophage (AM) product or products may interact with neutrophil elastase (NE) and modulate its binding to elastin. Elastase binding to elastase was evaluated by a modified elastase functional assay using a synthetic substrate. Supernatants from cultured AM inhibited elastase binding to elastin at a dose-dependent manner without inhibiting functional elastase activity. The AM products had a heterogeneous molecular weight ranging from 440,000 to 54,000. The activity was heat-stable, but was lost after ultracentrifugation. After lipid fractionation, neither the aqueous nor the lipid fractions contained activity, suggesting that the factor may be a lipid complex. Culture supernatants from smokers' AM released significantly higher amounts of the factor than nonsmokers. In addition, high molecular-weight elastase was present in bronchoalveolar lavage fluid (BALF) obtained from patients with pneumonia. Most of the in vivo high-molecular-weight elastase was lost after lipid extraction. In conclusion, macrophages release a factor or factors, probably lipid, which can interact with NE and inhibit its binding to human lung elastin without inhibiting elastase activity. This macrophage-derived factor may play a role in protecting the lung from NE by partitioning elastase into the airspace and thus protecting the interstitial connective tissue matrix from elastase degradation. PMID- 10471601 TI - Chest wall hyperinflation during acute bronchoconstriction in asthma. AB - The mechanics of the chest wall was studied in seven asthmatic patients before and during histamine-induced bronchoconstriction (B). The volume of the chest wall (VCW) was calculated by three-dimensional tracking of 89 chest wall markers. Pleural (Ppl) and gastric (Pga) pressures were simultaneously recorded. VCW was modeled as the sum of the volumes of the pulmonary-apposed rib cage (VRC,p), diaphragm-apposed rib cage (VRC,a), and abdomen (VAB). During B, hyperinflation was due to the increase in end-expiratory volume of the rib cage (0.63 +/- 0.09 L, p < 0.01), whereas change in VAB was inconsistent (0.09 +/- 0.07 L, NS) because of phasic recruitment of abdominal muscles during expiration. Changes in end-expiratory VRC,p and VRC,a were along the rib cage relaxation configuration, indicating that both compartments shared proportionally the hyperinflation. VRC,p Ppl plot during B was displaced leftward of the relaxation curve, suggesting persistent activity of rib cage inspiratory muscles throughout expiration. Changes in end-expiratory VCW during B did not relate to changes in FEV(1) or time and volume components of the breathing cycle. We concluded that during B in asthmatic patients: (1) rib cage accounts largely for the volume of hyperinflation, whereas abdominal muscle recruitment during expiration limits the increase in VAB; (2) hyperinflation is influenced by sustained postinspiratory activity of the inspiratory muscles; (3) this pattern of respiratory muscle recruitment seems to minimize volume distortion of the rib cage at end-expiration and to preserve diaphragm length despite hyperinflation. PMID- 10471602 TI - Bronchodilator effect of inhaled olprinone, a phosphodiesterase 3 inhibitor, in asthmatic patients. AB - The effect of topical administration of phosphodiesterase (PDE) 3 inhibitors on the airway is not clear. In order to examine the usefulness of inhaled PDE3 inhibitors in the treatment of asthma, we investigated the bronchodilator effect of inhaled olprinone, a newly developed PDE3 inhibitor, in nine asthmatic patients. On three separate study days, olprinone, salbutamol, or vehicle was administered in a double-blind and randomized fashion, and pulmonary functions were assessed over 60 min. Significant increases in FEV(1) were observed until 45 min after inhalation of olprinone without adverse cardiovascular effects. Mean maximal increases in FEV(1) were 16.0 +/- 4.0 and 20.5 +/- 4.2% with olprinone and salbutamol, respectively. The bronchodilator effect of olprinone was greater than that of salbutamol in four of the nine patients. These results suggest that the inhaled PDE3 inhibitor has a bronchodilator effect in asthmatic patients. PMID- 10471603 TI - Model of functional restriction in chronic obstructive pulmonary disease, transplantation, and lung reduction surgery. AB - Mechanical interactions between lung and chest wall are important determinants of respiratory function. When chest wall expansion during maximal inhalation generates insufficiently negative pleural pressures, the lungs remain functionally underinflated; this may be termed functional restriction. To explore mechanisms and effects of functional restriction in patients with emphysema, and to predict effects of single lung transplantation and lung volume reduction surgery (LVRS), we used a computational model based on standard physiology and measurements from individual patients. The model's lungs, separated by a compliant mediastinum, exhibit flow limitation according to the equal pressure point approach of Mead and coworkers. Pulmonary elastic recoil pressure is characterized by an exponential equation modified to reflect airway closure. Simulated respiratory maneuvers can be specified by variations in flow or pressure at the airway opening or in respiratory muscle activation. Model simulations successfully mimic recordings from individual patients. Input parameter values may then be altered to predict effects of surgical interventions in these same patients. The model simulations show the following. Single lung transplantation in emphysema can cause functional restriction of the normal transplanted lungs, and larger transplanted lungs may perform less well than smaller ones. LVRS improves lung and chest wall function in emphysema, but not in normal states. Surgical reduction of the native emphysematous lung after single lung transplantation can reduce functional restriction of the transplant and thereby improve its function. PMID- 10471604 TI - Ribavirin in ventilated respiratory syncytial virus bronchiolitis. A randomized, placebo-controlled trial. AB - We undertook a prospective, double-blind, placebo-controlled trial to resolve the question of the clinical effectiveness of ribavirin in previously well infants who require ventilation for respiratory distress secondary to respiratory syncytial virus (RSV) bronchiolitis. Aerosol ribavirin or NaCl 0.9% was administered within 24 h of initiation of ventilation, 18 h/d, for a maximum of 7 d or until extubation. From March 1994 to March 1997, 42 children were randomized and 41 patients were retained for analysis. Baseline characteristics of each group-ribavirin and placebo (20:21)-were not significantly different with respect to age (62.5 +/- 35.9 versus 62.7 +/- 30.9 d), sex, weight, and length of ventilation pre-aerosol. "Intent to treat" outcome analysis found no significant differences in the length of the following: ventilation (102.16 +/- 65.26 versus 126.28 +/- 78.72 h; p = 0.29), aerosol therapy, stay in the intensive care unit, total oxygen therapy, and hospitalization. The aerosols were well tolerated and no deaths occurred. This trial demonstrates the lack of effectiveness of aerosolized ribavirin in reducing the length of ventilation and course of illness in infants with no underlying illness ventilated for respiratory distress secondary to RSV bronchiolitis. PMID- 10471605 TI - Rate constant for forced expiration decreases with lung growth during infancy. AB - Airway caliber and lung volume (VL) increase many fold between infancy and adulthood; however, these two components of the lung may not increase proportionately during lung growth and development. We evaluated in infants the rate of emptying during forced expiration from near total lung capacity to residual volume. From the flow-volume curves we calculated (1) a rate constant (k) as the change in flow divided by the change in volume between 50% and 75% of expired forced vital capacity (FVC), and (2) the fraction of the FVC expired in 0.5 s (FEV(0.5)/FVC). Seventeen normal healthy infants were evaluated twice; mean ages (ranges) at first and second tests were 30 (5 to 76) and 58 (28 to 98) wk. Analysis of cross-sectional and longitudinal data indicated that the rate of emptying during forced expiration measured by both parameters was greatest in the youngest infants and decreased during infancy. Our findings are consistent with the concept that younger infants have large airways relative to their VL and that VL increases more rapidly than airway caliber early in life. PMID- 10471606 TI - A dobutamine test can disclose hepatosplanchnic hypoperfusion in septic patients. AB - In 36 hemodynamically stable septic patients, we explored whether changes in gastric mucosal-arterial PCO(2) gradient (PCO(2)gap) induced by a short-term dobutamine infusion may reveal hepatosplanchnic hypoperfusion. Hepatosplanchnic blood flow (HSBF) was determined by the continuous indocyanine green infusion technique and gastric mucosal PCO(2) (Pg(CO(2))) by saline tonometry. In each patient, hemodynamic measurements, blood samples, and Pg(CO(2)) determinations were performed three times: first at baseline (DOB 0), second during a dobutamine infusion at a dose of 5 microgram/kg/min (DOB 5), and third at a dose of 10 microgram/kg/min (DOB 10). The results were analyzed by Wilcoxon's matched-pairs signed rank test and are presented as medians with ranges. The PCO(2)gap decreased preferentially in groups of patients with inadequate hepatosplanchnic perfusion, i.e., with a low fractional HSBF (HSBF/CI), defined as the ratio of the HSBF to the simultaneous cardiac index, or a high gradient between the mixed venous blood and the suprahepatic blood O(2) saturations (DSvh(O(2))). In the 11 patients with a DSvh(O(2)) above 20% at baseline, PCO(2)gap decreased from 12.1 (6.3 to 19.5) mm Hg at DOB 0 to 6.2 (2.5 to 19. 3) mm Hg at DOB 5 (p < 0.001 versus DOB 0), and to 4.2 (0.1 to 35.9) mm Hg at DOB 10 (p < 0.05 versus DOB 5), whereas in the 25 patients with a DSvh(O(2)) below 20% at baseline, PCO(2)gap did not change significantly. At no time was the PCO(2)gap correlated with HSBF/CI or DSvh(O(2)). We conclude that although the PCO(2)gap does not correlate well with global indexes of gut oxygenation, such a simple dobutamine infusion test could identify patients with inadequate hepatosplanchnic perfusion. PMID- 10471607 TI - Prostacyclin enhances stretch-induced surfactant secretion in alveolar epithelial type II cells. AB - Inhalative vasodilator therapy, employing gaseous nitric oxide (NO) or aerosolized prostaglandin PGI(2), is of interest for regional pulmonary vasodilation in ARDS and pulmonary hypertension. We investigated the impact of the NO donor spermine NONOate as well as PGI(2) and its stable chemical analog iloprost on cultured rat alveolar epithelial type II cell (ATII) surfactant secretion. The NO donor provoked a significant increase in the ATII cGMP content, further enhanced by type V phosphodiesterase (PDE) inhibition, but affected neither baseline nor mechanical stretch-induced surfactant secretion. The prostanoids caused a marked increase in the epithelial cAMP content, further amplified by coadministration of type III/IV PDE inhibitors. Baseline surfactant secretion was not altered by this approach, but mechanical stretch-induced liberation of surfactant was significantly increased, most prominently in the ATII with the highest cAMP levels due to the presence of both iloprost and PDE III/IV inhibitors. In contrast, epithelial phosphoinositide metabolism, well responsive to purinergic stimulation as positive control, was unchanged in prostanoid-exposed cells. We conclude that the PGI(2)-cAMP axis, but not the NO cGMP axis, forwards a markedly enhanced secretory response to the physiological stimulus of cell surface stretching, which may be relevant for therapeutic use of these agents. PMID- 10471608 TI - Impact of immunomodulating therapy on morbidity in patients with severe sepsis. AB - We assessed the impact, over a 28-d period, of therapy with the tumor necrosis factor (TNF) neutralizing receptor fusion protein (p55-IgG) on the incidence of end-organ failures in patients with severe sepsis or early septic shock in a subgroup of 165 patients recruited into a randomized, multicenter clinical trial to receive placebo (n = 78) or a single infusion of p55-IgG, 0.083 mg/kg (n = 87). At study entry, distribution of organ dysfunctions and other baseline characteristics were similar for the two study groups. Treatment with p55-IgG was associated with a trend toward reduced 28-d mortality (p = 0.07), a decreased incidence of new organ dysfunctions (relative risk [RR], 0.57; 95% confidence interval [95% CI] 0.29 to 1.10, p = 0.10), and a decreased overall incidence density of organ failures (RR 0.65; 95% CI 0.60 to 0.71, p = 0.0001). Patients treated with p55-IgG had more organ failure-free days after study entry than those who received placebo. Average intensive care unit (ICU) stay was 2.6 d shorter (95% CI 0.2 to 5.0) for patients who received p55-IgG than for those who received placebo. For those patients who survived, this difference was 4.1 d (95% CI 1.6 to 6.6). Duration of ventilatory support was 3.2 d shorter (95% CI 0.1 to 6.3) among 28-d survivors who received p55-IgG, compared with placebo. In conclusion, in the population of septic patients studied, treatment with p55-IgG was associated with a trend toward shorter need for mechanical ventilatory support, a decreased length of stay (LOS), and a decreased incidence and duration of organ failure. PMID- 10471609 TI - The cGMP-specific phosphodiesterase inhibitor E4021 dilates the pulmonary circulation. AB - We investigated the pulmonary vascular effects of E4021, a potent inhibitor of cGMP-specific phosphodiesterase, in control late-gestation fetal lambs, and in newborn lambs with persistent pulmonary hypertension (PPHN) after prenatal ligation of the ductus arteriosus. E4021 alone significantly relaxed fifth generation pulmonary arteries isolated from control fetal lambs, an effect completely blocked after inhibition of nitric oxide synthase (NOS). In contrast, E4021 did not relax pulmonary arteries isolated from hypertensive lambs. Pretreatment with E4021 (10(-7) M) significantly enhanced relaxations to the NO donor S-nitrosyl-acetyl-penicilamine (SNAP) in arteries from both control and hypertensive lambs. In control, fully instrumented fetal lambs, infusions of E4021 (31 microgram/min) selectively dilated the pulmonary circulation, an effect again blocked after inhibition of NO synthase. Further studies were performed in newborn lambs with PPHN to study the vascular effects of E4021 alone, and in combination with inhaled NO. E4021 alone (1 to 100 microgram/kg/min) decreased pulmonary artery pressure (Ppa) in a dose-dependent fashion, and had minimal effect on systemic pressure. At the highest dose (100 microgram/kg/min), the dilation was selective for the pulmonary circulation. In subsequent protocols, E4021 (10 microgram/kg/min) significantly decreased Ppa and pulmonary vascular resistance (PVR), but these pulmonary vascular effects were not enhanced after NO inhalation at 0.5 or 5 ppm. We speculate that the lack of enhancement was due to the dramatic effects of E4021 alone. Potent, specific phosphodiesterase inhibitors such as E4021 may prove to be useful in the treatment of PPHN. PMID- 10471610 TI - Effect of clarithromycin regimen for Mycobacterium avium complex pulmonary disease. AB - We have investigated the efficacy of a clarithromycin-containing four-drug regimen for Mycobacterium avium complex (MAC) pulmonary disease in 46 patients without acquired immunodeficiency syndrome (AIDS). The patients were 14 males and 32 females with a mean age of 60.9 +/- 11.5 yr. Patients received 10 mg/kg/d of clarithromycin plus ethambutol, rifampin, and initial kanamycin and subsequent quinolone for 24 mo. Seven patients (15.2%) were dropped in the first 6 mo. Among 39 patients who received more than 6 mo of therapy, 28 patients (71.8%) converted their sputa to negative: 26 of 31 patients (83.9%) infected with clarithromycin susceptible strains and two of eight patients (25.0%) with resistant or intermediate strains. The timing of sputum conversion was 3.6 +/- 1.9 mo, with a range of 2 to 9 mo. The conversion rate was significantly lower in patients who were infected with clarithromycin-resistant or intermediate strains, who had had prior therapy (55.0% versus 89.5%), or who were acid-fast bacilli (AFB) smear positive at entry (60.7% versus 100%). The age and sex of patients, the species of pathogen (M. avium or M. intracellulare), type and extent of the disease, and the use of kanamycin did not significantly affect the conversion rate. Although the regimen was efficacious for newly treated patients, frequent adverse reactions and a low conversion rate of sputum in retreated patients are problems that remain to be solved. PMID- 10471611 TI - Interleukin-18 enhances antigen-induced eosinophil recruitment into the mouse airways. AB - Interleukin-18 (IL-18) has recently been identified as an IFN-gamma-inducing factor. Previous studies have shown that CD4(+) T cells, IL-5, and TNF-alpha mediate, but IFN-gamma and IL-12 (via IFN-gamma production) inhibit antigen induced eosinophil recruitment into the airways of sensitized mice. Here, we showed that the administration of recombinant murine IL-18 enhanced antigen induced eosinophil recruitment into the trachea and bronchoalveolar lavage fluids (BALF) of sensitized mice in a dose-dependent manner. The administration of IL-18 enhanced antigen-induced IFN-gamma and TNF-alpha production, but not IL-5 production, in the BALF and lungs of sensitized mice. Neutralizing antibody against TNF-alpha prevented antigen-induced eosinophil recruitment into the BALF of sensitized mice. Although IL-18 enhanced antigen-induced airway eosinophilia, IL-18 did not affect antigen-induced airway hyperresponsiveness in sensitized mice. These results indicate that IL-18, unlike IFN-gamma and IL-12, enhances antigen-induced eosinophil recruitment into the airways in part by increasing antigen-induced TNF-alpha production of sensitized animals. These findings suggest that IL-18 may contribute to the development and exacerbation of airway inflammation in asthma. PMID- 10471612 TI - Exhaled nitric oxide does not provide a marker of vascular endothelial function in healthy humans. AB - In the lung, nitric oxide synthase (NOS) has been found in both alveolar epithelial and vascular endothelial cells. Nitric oxide (NO) in the exhaled air stemming from the lower respiratory tract has been claimed to represent a marker of the vascular endothelial NO production. Experimental evidence for this concept, however, is lacking. We compared, in eight healthy volunteers, effects on exhaled NO of epithelial NOS inhibition by N (G)-monomethyl-L-arginine (L NMMA) inhalation (6 mg/kg over 15 min) with those of endothelial NOS inhibition by L-NMMA infusion (25 microgram/kg/min for 30 min). We also measured blood pressure, heart rate, and L-NMMA plasma concentration. The major new findings were that L-NMMA inhalation which did not have any detectable effect on hemodynamics and L-NMMA plasma concentration, decreased the pulmonary exhaled NO by almost 40%. In contrast, L-NMMA infusion that inhibited endothelial NOS, as evidenced by an increase in blood pressure and a decrease in heart rate, had only a barely detectable effect on exhaled NO (-11 +/- 4% from baseline). Pulmonary exhaled NO is mostly of epithelial rather than endothelial origin, and does not provide a marker for vascular endothelial NO production and/or endothelial function in healthy humans. PMID- 10471613 TI - The impact of morbid obesity on oxygen cost of breathing (VO(2RESP)) at rest. AB - Oxygen consumption dedicated to respiratory work (V O(2RESP)) during quiet breathing is small in normal patients. In the morbidly obese, at high minute ventilations, VO(2RESP) is greater than in normal patients, but VO(2RESP) during quiet breathing in these patients is not known. We postulated that such patients have increased VO(2RESP) at rest which may predispose them to respiratory failure when additional respiratory workloads are imposed. We measured baseline VO(2) in morbidly obese patients immediately prior to gastric bypass surgery and again after intubation, mechanical ventilation, and paralysis, and compared their change in VO(2) to nonobese patients scheduled for elective abdominal surgery. Baseline VO(2) was higher in the obese patients compared with control patients (354.6 versus 221.4 ml/min; p = 0.0001) and the change in VO(2) from spontaneous breathing to mechanical ventilation was significant in the obese patients (354.6 versus 297.2 ml/min; p = 0.0002) but not the control patients (221.4 versus 219.8 ml/min; p = 0.86). We conclude that morbidly obese patients dedicate a disproportionately high percentage of total VO(2) to conduct respiratory work, even during quiet breathing. This relative inefficiency suggests a decreased ventilatory reserve and a predisposition to respiratory failure in the setting of even mild pulmonary or systemic insults. PMID- 10471614 TI - Acute asthma among pregnant women presenting to the emergency department. AB - Asthma complicates up to 4% of pregnancies. Our objective was to compare emergency department (ED) visits for acute asthma among pregnant versus nonpregnant women. We performed a prospective cohort study, as part of the Multicenter Asthma Research Collaboration. ED patients who presented with acute asthma underwent a structured interview in the ED, and another by telephone 2 wk later. The study was performed at 36 EDs in 18 states. A total of 51 pregnant women and 500 nonpregnant women, age 18 to 39, were available for analysis. Pregnant women did not differ from nonpregnant women by duration of asthma symptoms (median: 0.75 versus 0.75 d, p = 0.57) or initial peak expiratory flow rate (PEFR) (51% versus 53% of predicted, p = 0.52). Despite this similarity, only 44% of pregnant women were treated with corticosteroids in the ED compared with 66% of nonpregnant women (p = 0.002). Pregnant women were equally likely to be admitted (24% versus 21%, p = 0.61) but less likely to be prescribed corticosteroids if sent home (38% versus 64%, p = 0.002). At 2-wk follow-up, pregnant women were 2.9 times more likely to report an ongoing exacerbation (95% CI, 1.2 to 6.8). Among women presenting to the ED with acute asthma, pregnant asthmatics are less likely to receive appropriate treatment with corticosteroids. PMID- 10471615 TI - The interrelationship of sputum inflammatory markers in patients with chronic bronchitis. AB - Many of the features of bronchial disease are believed to be caused by damage to the airways by elastase released by recruited neutrophils. There have been few studies of the mechanisms involved and the interrelationships between components of the inflammatory process. We studied secretions from patients with chronic bronchitis in the stable state. We assessed the presence of neutrophils by measuring myeloperoxidase (MPO) activity and active neutrophil elastase (NE). These results were compared with the chemoattractants interleukin-8 (IL-8) and leukotriene B(4) (LTB(4)), the bronchial inhibitor secretory leukoprotease inhibitor (SLPI), and protein leak (sputum/serum albumin ratio). MPO correlated with NE activity (r = 0.68, p < 0.001) and both IL-8 (r = 0.52, p < 0.001) and LTB(4) (r = 0.41, p < 0.001) indicating an association with the chemoattractants. Elastase activity correlated with IL-8 (r = 0.55, p < 0.001) and LTB(4) (r = 0.41, p < 0.001) but negatively with SLPI (r = -0.49, p < 0.001). NE also correlated positively with protein leak (r = 0.36, p < 0.001), suggesting a cause and effect. MPO and protein leak correlated negatively with FEV(1) (percentage of predicted) only in patients with chronic obstructive pulmonary disease (COPD) without alpha(1)-antitrypsin deficiency (r = -0.37, p < 0.001; r = -0.42, p < 0.01, respectively). These complex interactions provide a template for future studies with specific inhibitors or agonists which will clarify the role of individual factors. PMID- 10471616 TI - A histologic pattern of nonspecific interstitial pneumonia is associated with a better prognosis than usual interstitial pneumonia in patients with cryptogenic fibrosing alveolitis. AB - This study aimed to investigate whether there was a difference in outcome related to histologic pattern in cryptogenic fibrosing alveolitis (CFA) and to see whether there were correlations between clinical and radiologic findings and histology. One hundred thirteen lung biopsies from consecutive patients taken for the diagnosis of diffuse lung disease were reviewed and reclassified using the Katzenstein and Myers criteria for interstitial pneumonias. Patients lacking full investigational data at presentation and those with conditions predisposing to lung fibrosis were excluded, leaving 15 patients diagnosed with nonspecific interstitial pneumonia (NSIP) and 15 with usual interstitial pneumonia (UIP). Clinical and radiologic findings at presentation and serial lung function information and survival status in November 1998 were compared for the two groups. Survival was found to be significantly greater in the NSIP group compared with the UIP group (p < 0.001). This could not be explained by differences in treatment. Patients with UIP showed a progressive deterioration in lung function whereas those with NSIP remained stable. CT scans of patients with UIP showed more fibrosis than those of patients with NSIP (p < 0.011). A histologic diagnosis of NSIP is associated with a better prognosis than UIP. This subclassification of CFA is clinically useful. PMID- 10471617 TI - Pulmonary rapidly adapting receptor stimulation does not increase airway resistance in anesthetized rabbits. AB - In open-chest artificially ventilated rabbits, removal followed by replacement of positive end-expiratory pressure (PEEP maneuver) favors stimulation of airway rapidly adapting receptors (RARs). The purpose of the present study was to determine whether activation of RARs can cause bronchoconstriction. We measured airway pressure, airflow, and tidal volume, and calculated dynamic lung compliance and total lung resistance. PEEP maneuver increased airway pressure swings (16.4 +/- 4% above control; p = 0.0016) and decreased compliance (to 84.8 +/- 2.8% of control; p = 0.0002) without changing resistance (108.0 +/- 4.4% of control; p = 0.85). On the other hand, the resistance increased greatly (93 +/- 13%, p < 0.01) after intravenous injection of acetylcholine or electrical stimulation of vagal efferents, indicating that our system could detect increases in the resistance. In a separate group, we stimulated RARs by stroking the trachea with a cotton tip (tickling), tickling produced cough, manifested by increased pressure and flow without resistance changing. These changes were abolished after paralysis with succinylcholine. Because we did not detect an increase in airflow resistance during activation of RARs by the PEEP maneuver and tickling, we conclude that increase in resistance may not be an important reflex component of airway RARs. PMID- 10471618 TI - Noninvasive diagnosis of emphysema. Aerosol morphometry and aerosol bolus dispersion in comparison to HRCT. AB - Aerosol-derived airway morphometry (ADAM) and aerosol bolus dispersion (ABD) test are altered in patients with emphysema. We examined the diagnostic power of these aerosol methods in comparison with the noninvasive "gold-standard" HRCT in 50 consecutive patients with various lung diseases. The severity of airflow limitation was mild to moderate in the group of patients without emphysema and moderate to severe in the group of patients with HRCT-confirmed emphysema (FEV(1), 78 +/- 23% pred versus 53 +/- 33% pred; p < 0. 001). Among all lung function parameters under consideration ADAM showed the highest sensitivity and specificity for separating patients with emphysema from those without emphysema (area under the operating characteristics curve: p(ROC), 0.92), followed by ABD (p(ROC), 0.90), a marker for ventilation inhomogeneities. In patients with HRCT confirmed macroscopic emphysema, peripheral air-space dimensions (EAD) at a relative volumetric lung depth V(pr) of 0.20 measured by ADAM were 155% larger, and bolus dispersion (ABD) at a lung depth of V(p) 600 ml was 53% larger than those observed in patients with other lung diseases (EAD = 0.84 +/- 0.53 mm versus 0.33 +/- 0.10 mm, p < 0.0001; ABD = 706 +/- 154 cm(3) versus 462 +/- 109 cm(3); p < 0.0001). EAD showed a significant correlation with the HRCT visual score (r = 0.78, p = 0.01). ABD showed weak significant correlations with all HRCT parameters under consideration (visual score, pixel density, mean lung density) (r = 0.45 to 0.66; p < 0.05). ADAM and ABD are powerful tools for the noninvasive diagnosis of macroscopic emphysema. PMID- 10471619 TI - Association between a sequence variant in the IL-4 gene promoter and FEV(1) in asthma. AB - Recent family-based studies have revealed evidence for linkage of human chromosome 5q31 to the diagnosis of asthma, elevated serum IgE levels, and bronchial hyperresponsiveness. Among the candidate genes in this region is the gene encoding for human interleukin-4 (IL-4). We reasoned that this gene could also serve as a candidate gene with respect to asthma severity as indicated by the FEV(1) measured when bronchodilator treatment was withheld. To test this hypothesis, we examined a large population of patients with asthma (ascertained without respect to genetic characteristics), for associations between a genetic variant in the IL-4 promoter region (C-589T) and asthma severity, as indicated by FEV(1). We used amplification by the polymerase chain reaction followed by BsmF1 restriction digestion to assign genotypes at the IL-4 promoter C-589T locus. We compared genotypes at this locus in 772 Caucasian and African American patients with asthma of varying severity, and we used multiple regression analysis to relate genotypic findings to FEV(1). Among white individuals, the homozygous presence of the C-589T IL-4 promoter genotype (TT) was associated with a FEV(1) below 50% of predicted (p = 0.013; OR, 1.44; 95% CI: 1.09 to 1.90). Subjects with the TT genotype had mean FEV(1) (% predicted) values 4.5% lower than those of subjects with the wild-type (CC) genotype at this locus. FEV(1) values of white patients with a CC or CT genotype were broadly distributed, whereas the TT genotype was associated with a narrow distribution of low FEV(1) values. The frequency of the T allele was significantly greater (p = 1 x 10(-)(23)) among African American asthmatics (0.544) than among white asthmatics (0.183). These data provide the first evidence associating FEV(1) in patients with asthma and genetic determinants at any locus. Our data are consistent with the idea that the FEV(1) in asthma is the result of multiple factors; one of these factors is the genotype at the IL-4 C-589T locus. This locus is associated with a small but significant decrement in pulmonary function among white asthmatic subjects. PMID- 10471620 TI - Severe community-acquired pneumonia. Risk factors and follow-up epidemiology. AB - The aim of the study was to determine risk factors for severe community-acquired pneumonia (CAP) as well as to compare microbial patterns of severe CAP to a previous study from our respiratory intensive care unit (ICU) originating from 1984 to 1987. Patients admitted to the ICU according to clinical judgment were defined as having severe CAP. For the study of risk factors, a hospital-based case-control design was used, matching each patient with severe CAP to a patient hospitalized with CAP but not requiring ICU admission. Microbial investigation included noninvasive and invasive techniques. Overall, 89 patients with severe CAP were successfully matched to a control patient. The presence of an alcohol ingestion of >/= 80 g/d (odds ratio [OR] 3.9, 95% confidence interval [CI] 1.4 to 10.6, p = 0.008) was found to be an independent risk factor for severe CAP and prior ambulatory antimicrobial treatment (OR 0.37, 95% CI 0.17 to 0.79, p = 0.009) to be protective. Streptococcus pneumoniae (24%) continued to be the most frequent pathogen; however, 48% of strains were drug-resistant. "Atypical" bacterial pathogens were significantly more common (17% versus 6%, p = 0.006) and Legionella spp. less common (2% versus 14%, p = 0.004) than in our previous study, whereas gram-negative enteric bacilli (GNEB) and Pseudomonas aeruginosa continued to represent important pathogens (6% and 5%, respectively). Our findings provide additional evidence for the importance of the initiation of early empiric antimicrobial treatment for a favorable outcome of CAP. Variations of microbial patterns are only in part due to different epidemiological settings. Therefore, initial empiric antimicrobial treatment will also have to take into account local trends of changing microbial patterns. PMID- 10471621 TI - Clara cell protein-positive epithelial cells are reduced in small airways of asthmatics. AB - Clara cell 10 kilodalton protein (CC10), the predominant product from nonciliated cells in the epithelial lining of bronchioles (Clara cells), has been shown to have immunomodulatory and anti-inflammatory activity, and may play roles in controlling inflammation in the airway. This study was designed to examine immunohistochemical expression of CC10 in epithelial cells in small airways (perimeter < 6 mm) of asthmatic and control nonsmokers who underwent lung resection because of peripheral lung carcinoma and to compare CC10-positive epithelial cell proportions with numbers of inflammatory cells in small airways of asthmatics. Significantly decreased proportions of CC10-positive epithelial cells and significantly increased numbers of T cells, activated eosinophils, and mast cells in small airways of asthmatics were found compared with those of control subjects. CC10-positive epithelial cell proportions inversely correlated with numbers of T cells and mast cells in small airways of asthmatics. Decreases of CC10-producing cells may give an accelerating cause for further aggravation of inflammatory responses in chronic asthma. PMID- 10471622 TI - Anti-interleukin 5 but not anti-IgE prevents airway inflammation and airway hyperresponsiveness. AB - The role of IL-5 and allergen-specific IgE in the development of eosinophilic airway inflammation and airway hyperresponsiveness (AHR) was investigated in a murine model. BALB/c mice were sensitized to ovalbumin (OVA) by intraperitoneal injection on Days 1 and 14, followed by airway challenge with OVA on Days 28 and 29. Anti-IL-5 (TRFK-5) or anti-IgE (antibody 1-5) was administered before each airway challenge. Sensitized and challenged mice developed increased OVA-specific IgE serum levels, Th2 cytokine production by peribronchial lymph node (PBLN) cells, increased numbers of eosinophils (predominantly located in the peribronchial regions of the lungs), and increased airway responsiveness to methacholine (MCh). Anti-IgE treatment significantly decreased serum anti-OVA IgE levels and prevented the development of anaphylaxis but failed to affect T cell function, eosinophil airway infiltration, and AHR in sensitized and challenged mice. In contrast, treatment with anti-IL-5 antibody did not affect B cell (Ig serum levels), T cell (cytokine production), or mast cell function (immediate cutaneous reactivity) but completely inhibited development of eosinophilic lung inflammation and AHR. These data identify IL-5-mediated eosinophilia as a major target for development of AHR in this model, with little effect resulting from neutralization of IgE. PMID- 10471623 TI - Surfactant proteins A and D in premature baboons with chronic lung injury (Bronchopulmonary dysplasia). Evidence for an inhibition of secretion. AB - Surfactant proteins A and D (SP-A and SP-D) are believed to participate in the pulmonary host defense and the response to lung injury. In order to understand the effects of prematurity and lung injury on these proteins, we measured the amounts of SP-A and SP-D and their mRNAs in three groups of animals: (1) nonventilated premature baboon fetuses; (2) neonatal baboons delivered prematurely at 140 d gestation age (ga) and ventilated with PRN O(2); (3) animals of the same age ventilated with 100% O(2) to induce chronic lung injury. In nonventilated fetuses, tissue and lavage SP-A were barely detectable in baboons of 125 and 140 d ga, but they equaled or exceeded adult SP-A concentrations (g/g lung dry wt) at 175 d (term gestation, 185 d). In contrast, SP-D was readily detectable in tissue and lavage at 125 and 140 d ga. When the baboons of 140 d ga were ventilated for 10 d with 100% oxygen to produce chronic lung injury, the tissue concentration of SP-A was five times greater than that of normal adults; SP-D 16-times greater. Despite the sizable tissue pools of SP-A and SP-D, however, lavage SP-A was only 7% of that of normal adults and lavage SP-D just equaled the amount in normal adults. Nevertheless, because SP-D is normally in much lower concentration than is SP-A, their total comprised less than 12% of the SP-A and SP-D found in the lavage of a healthy adult. The results indicate that in chronic lung injury, SP-A is significantly reduced in the alveolar space. SP-D concentration in lavage is about equal to that in normal adults, possibly because of the 16-fold excess in tissue, but the total collectin pool in lavage is still significantly reduced. Because these collectins may bind and opsonize bacteria and viruses, decrements in their amounts may present additional risk to those premature infants who require prolonged periods of ventilatory support. PMID- 10471624 TI - Entrainment of respiration in humans by periodic lung inflations. Effect of state and CO(2). AB - Lack of synchrony between a patient and the mechanical ventilator occurs when the respiratory rhythm of the patient fails to entrain to machine inflations. Entrainment implies a resetting of the respiratory rhythm such that a fixed temporal relationship exists between the onset of inspiratory activity and the onset of a mechanical breath. We examined the entrainment response to mechanical ventilation of normal humans over a range of machine rates during wakefulness and during isocapnic and hypercapnic NREM sleep. Wakefulness facilitated 1:1 entrainment of the respiratory rhythm to the mechanical ventilator over a wider range of machine frequencies than during NREM sleep (p < 0.001); isocapnic and hypercapnic conditions did not differ (p = 0.95). To evaluate the Hering-Breuer reflexes in the resetting of the respiratory rhythm during sleep, we examined changes in neural inspiratory time (TI) as the relationship between inspiratory efforts and onset of machine inflations changed. As inspiratory efforts extended into the machine inflation cycle, neural TI shortened. We conclude that entrainment responses of normal humans to mechanical ventilation differ depending on state, but mild increases in respiratory drive caused by CO(2) stimulation do not affect these entrainment responses. Furthermore, the changes in neural TI are consistent with observations in animal studies in which Hering-Breuer reflexes mediated entrainment. PMID- 10471625 TI - Cytokines IL-1beta, IL-6, and TNF-alpha enhance in vitro growth of bacteria. AB - We have previously reported that in acute respiratory distress syndrome (ARDS), nonsurvivors have persistent elevation in pulmonary and circulating proinflammatory cytokine levels over time and a high rate of nosocomial infections antemortem. In these patients, none of the proven or suspected nosocomial infections caused a transient or sustained increase in plasma proinflammatory cytokine levels above preinfection values. We hypothesized that cytokines secreted by the host during ARDS may favor the growth of bacteria. We conducted an in vitro study of the growth of three bacteria clinically relevant in nosocomial infections, evaluating their in vitro response to various concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-6. We found that all three bacterial species showed concentration-dependent growth enhancement when incubated with one or more tested cytokines and that blockade by specific neutralizing cytokine MoAb significantly inhibited cytokine induced growth. When compared with control, the 6-h growth response (cfu/ml) was maximal with IL-1beta at 1,000 pg for Staphylococcus aureus (36 +/- 16 versus 377 +/- 16; p = 0.0001) and Acinetobacter spp. (317 +/- 1,147 versus 1,124 +/- 147; p = 0.002) and with IL-6 at 1,000 pg for Pseudomonas aeruginosa (99 +/- 50 versus 509 +/- 50; p = 0.009). The effects of cytokines were seen only with fresh isolates and were lost with passage in vitro on bacteriologic medium without added cytokines. In this study we provide additional evidence for a newly described pathogenetic mechanism for bacterial proliferation in the presence of exaggerated and protracted inflammation. PMID- 10471626 TI - Elastosis and fragmentation of fibers of the elastic system in fatal asthma. AB - Fibers of the elastic system (FES) are important in regulating airway patency and lung elastic recoil. Their possible role in modulating bronchoconstriction is not fully understood. Chronic inflammation and mechanical stretching present in asthma could lead to conformational alterations in the FES, participating in the mechanism of airway remodeling observed in this disease. In airway mucosa, two layers of FES are discernible: one superficial network, attached to the basement membrane, and a deeper network, lying close to the airway smooth muscle. Using image analysis and conventional morphometry, we analyzed airway content of FES in central and peripheral airways in 31 cases of fatal asthma and in 10 control lungs. Slides were stained using the oxidized resorcin-fuchsin method. Optical analysis revealed fragmentation of the FES in the superficial network of central asthmatic airways. Morphometry showed the presence of elastosis in central asthmatic airways when the entire thickness of the mucosa was considered. In the superficial network elastic fiber content was significantly decreased. These results indicate that FES participate in airway remodeling in asthma. We also suggest that disrupture of fiber attachments at the basement membrane in the superficial layer could impair the mechanism of airway recoil in asthmatic patients. PMID- 10471627 TI - The attributable mortality and costs of primary nosocomial bloodstream infections in the intensive care unit. AB - Primary nosocomial bloodstream infection (BSI) is a common occurrence in the intensive care unit (ICU) and is associated with a crude mortality of 31.5 to 82.4%. However, an accurate estimate of the attributable mortality has been limited because of confounding by severity of illness. We undertook this study to assess the attributable mortality and costs associated with an episode of BSI. Infected patients were defined as those who had an episode of BSI during the study period. Uninfected control subjects were matched to the infected patients based upon a number of factors, including predicted mortality on the day prior to infection. The main outcome measures were crude ICU mortality, length of stay, and costs. We found no difference in the crude mortality for the infected and the uninfected patients (35.3 and 30.9%, respectively, p = 0.51). However, among survivors, the patients with nosocomial bloodstream infections did have excess length of stay (mean, 10 d; median, 5 d; p = 0.007) and increased direct costs (mean difference, $34,508; p = 0.008). After matching for severity of illness, we could not detect an association between primary nosocomial bloodstream infections and increased ICU mortality. We did find that primary nosocomial bloodstream infections increased ICU length of stay and costs. PMID- 10471628 TI - Stimulation of the dopamine 1 receptor increases lung edema clearance. AB - We previously reported that lung edema clearance was stimulated by dopamine (DA). The purpose of this study was to determine whether the DA-mediated stimulation of edema clearance occurs via an adrenergic or dopaminergic regulation of alveolar epithelial Na, K-ATPase. When isolated perfused rat lungs were coinstilled with DA and SCH 23390 (a specific D(1) receptor antagonist), there was a dose dependent attenuation of the stimulatory effects of DA. Coinstillation with S sulpiride (a specific D(2) receptor antagonist) or propranolol (a beta-adrenergic antagonist) did not alter DA-stimulated clearance. Similarly, the specific dopaminergic D(1) agonist fenoldopam increased lung edema clearance, but quinpirole (a specific dopaminergic D(2) agonist) did not. (125)I-SCH 23982 binding studies suggested that D(1) receptors are expressed on alveolar type II (ATII) cells with an apparent dissociation constant (K(d)) of 4.4 nM and binding maximum (Bmax) 9.8 pmol/mg. Consistent with these results, the D(1) receptor messenger RNA (mRNA) and protein were detected in ATII cells by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. These data demonstrate a novel mechanism involving the activation of dopaminergic D(1) receptors which mediates DA-stimulated edema removal from rat lungs. PMID- 10471629 TI - Antimyeloperoxidase-associated lung disease. An experimental model. AB - The lung is a common target in systemic vasculitides associated with antineutrophil cytoplasmic antibodies (ANCA). In the present study, we tested the hypothesis that the presence of antibodies directed against myeloperoxidase (MPO) induces pulmonary (vasculitic) lesions when neutrophils release lysosomal enzymes. Brown Norway (BN) rats were immunized with human MPO in complete Freund's adjuvant (CFA) or with CFA alone. Two weeks after immunization, rats had developed antibodies to human and rat MPO. Next, isolated single left lung perfusion was performed with human neutrophil lysosomal extract containing MPO and proteolytic enzymes. Rats were killed at 15 min, 4 h, and 10 d after perfusion. Tissue samples from the left and right lung were examined for vasculitic lesions and inflammatory cell infiltrates. At 15 min and 4 h, left lungs from control and MPO-immunized rats showed a mild influx of polymorphonuclear cells. At 10 d, patchy inflammatory cell infiltrates, consisting predominantly of polymorphonuclear leukocytes (PMNs) and monocytes, were observed throughout the parenchyma of the left lung in MPO-immunized rats. Occasionally, granuloma-like lesions, giant cells, and foci of alveolar hemorrhage were observed as well. Far less severe lesions were seen in control immunized rats. Strikingly, at 10 d after perfusion, severe pulmonary tissue injury was observed also in right lungs from MPO-immunized rats whereas right lungs from control immunized rats appeared normal. The lesions were characterized by influx of PMNs and monocytes and, in some rats, foci of alveolar hemorrhage. These studies suggest that the presence of an anti-MPO directed autoimmune response contributes to generalized pulmonary tissue injury after local release of products of activated neutrophils, which supports a pathogenic role of MPO ANCA. PMID- 10471630 TI - Induction of lymphocytic inflammatory changes in lung interstitium by human T lymphotropic virus type I. AB - Human T lymphotropic virus type I (HTLV-I) is the etiological agent of adult T cell leukemia, and reports suggest that several other clinical conditions are associated with HTLV-I infection, including myelopathy and inflammatory pulmonary diseases. However, the clinical entity of HTLV-I-associated lung disease remains unsubstantiated more than 10 years after its description. In the present study, we conducted a histopathological analysis of lung tissues of transgenic mice that expressed gene segments of HTLV-I p40(tax) regions. The aim of the study was to examine the relationship between expression of viral components and development of lung disorders. In these mice, inflammatory changes with infiltration of lymphocytes in peribronchial and perivascular areas and in alveolar septa developed at 11 wk of age and increased in incidence during the observation period (26 wk). There was a significant correlation between the pulmonary pathological changes and the level of expression of p40(tax) mRNA in the lungs. Our results provided for the first time strong evidence of a direct relationship between HTLV-I and development of bronchopulmonary infection. PMID- 10471631 TI - Evidence that severe asthma can be divided pathologically into two inflammatory subtypes with distinct physiologic and clinical characteristics. AB - The mechanisms associated with the development of severe, corticosteroid (CS) dependent asthma are poorly understood, but likely heterogenous. It was hypothesized that severe asthma could be divided pathologically into two inflammatory groups based on the presence or absence of eosinophils, and that the inflammatory subtype would be associated with distinct structural, physiologic, and clinical characteristics. Thirty-four severe, refractory CS-dependent asthmatics were evaluated with endobronchial biopsy, pulmonary function, allergy testing, and clinical history. Milder asthmatic and normal control subjects were also evaluated. Tissue cell types and subbasement membrane (SBM) thickness were evaluated immunohistochemically. Fourteen severe asthmatics [eosinophil (-)] had nearly absent eosinophils (< 2 SD from the normal mean). The remaining 20 severe asthmatics were categorized as eosinophil (+). Eosinophil (+) severe asthmatics had associated increases (p < 0.05) in lymphocytes (CD3+, CD4+, CD8+), mast cells, and macrophages. Neutrophils were increased in severe asthmatics and not different between the groups. The SBM was significantly thicker in eosinophil (+) severe asthmatics than eosinophil (-) severe asthmatics and correlated with eosinophil numbers (r = 0.50). Despite the absence of eosinophils and the thinner SBM, the FEV(1) was marginally lower in eosinophil (-) asthmatics (p = 0.05) with no difference in bronchodilator response. The eosinophil (+) group (with a thicker SBM) had more intubations than the eosinophil (-) group (p = 0.0004). Interestingly, this group also had a decreased FVC/slow vital capacity (SVC). These results suggest that two distinct pathologic, physiologic, and clinical subtypes of severe asthma exist, with implications for further research and treatment. PMID- 10471632 TI - Analysis of MHC encoded antigen-processing genes TAP1 and TAP2 polymorphisms in sarcoidosis. AB - Sarcoidosis is a chronic granulomatous disease of unknown etiology. Several studies have suggested involvement of human leukocyte antigen (HLA) genes in sarcoidosis susceptibility. HLA associations described have not been consistent, possibly because of additional susceptibility genes adjacent to or within the major histocompatibility complex (MHC) such as genes for the transporter associated with antigen processing (TAP). The aim of this study was to analyze TAP gene polymorphisms in patients with sarcoidosis using the amplificatory refraction mutation system (ARMS) PCR. To determine whether any association between TAP gene variation and sarcoidosis was ethnic-independent we examined two European populations: 117 unrelated UK Caucasoid patients with sarcoidosis and 290 healthy UK control subjects, and 87 unrelated Polish Slavonic patients with sarcoidosis and 158 healthy Polish control subjects. We detected significant differences in TAP2 between the UK control and patient groups, and in TAP2 between the Polish control and patient groups. Comparing the UK and Polish control groups, we observed a difference in TAP1. Examination of HLA-DPB1 in our UK population showed no associations with disease or between variants at the TAP gene loci and HLA-DPB1 variants. These results suggest associations at the TAP loci occur independently of HLA-DPB1 associations, that TAP associations seen may be involved in determining sarcoidosis susceptibility, and that such susceptibilities differ between UK and Polish populations. This first study of TAP genes in UK and Polish sarcoid populations has demonstrated the importance of using multiple defined ethnic populations in defining the role genetic factors play in sarcoidosis susceptibility and the importance of candidate gene studies. PMID- 10471633 TI - Hyperacute rejection of a pulmonary allograft. Immediate clinical and pathologic findings. AB - The clinical and pathologic findings seen in hyperacute rejection are well documented in renal and cardiac allografts. We describe the second case of hyperacute rejection in a pulmonary allograft and detail the immediate clinicopathologic findings. The patient underwent a single lung transplant for severe COPD with postoperative course complicated by acute rejection and graft failure. Eleven days later, the patient underwent a second transplant with intra operative course complicated by rapid pulmonary edema and copious production of frothy, pink fluid from the bronchial orifice of the allograft followed by death within four hours of anastomoses. Intraoperative biopsy and autopsy demonstrated platelet/fibrin thrombi, marked interstitial neutrophilia, alveolar edema, and antibody deposition on the endothelial surface and vasculature walls. Prior to the first transplant, the patient's serum had 0% panel reactive antibody and was crossmatch compatible with the first allograft. The patient's serum prior to the second transplant contained cross-reacting antibodies to the donor's B and T lymphocytes. The immediate clinical findings in this case are similar to the findings in a previously reported case. This report is the first documentation of the immediate pathologic features of hyperacute rejection in a lung allograft which are similar to those seen with other organ allografts. PMID- 10471634 TI - Steroid-refractory neonatal eosinophilic pneumonia responsive to cyclosporin A. AB - Idiopathic neonatal eosinophilic pneumonia is extremely rare. We report an infant who presented with tachypnea and interstitial infiltrates on chest radiograph at age 2 wk. Lung biopsy revealed perivascular and interstitial eosinophils. Despite initial improvement, the patient's condition became resistant to corticosteroids, cromolyn, and intravenous gamma globulin. After treatment with cyclosporin A his symptoms resolved. PMID- 10471635 TI - Effect of aerosolized anti-IgE (E25) on airway responses to inhaled allergen in asthmatic subjects. AB - Intravenous administration of a humanized monoclonal antibody of IgE (E25) attenuates the early and late phase response to inhaled allergen in allergic asthmatic subjects. To test whether direct delivery of E25 to the airway might have the same effect, we conducted a randomized, double-blind, three group study in 33 subjects with mild allergic asthma (20 to 46 yr of age, 21 men, FEV(1) > 70% predicted). The airway responses to aerosolized allergen were determined at baseline, after 2 and 8 wk of once daily treatment with aerosolized placebo (n = 11), aerosolized E25 1 mg (n = 12), or aerosolized E25 10 mg (n = 10), and after 4 wk of treatment withdrawal. We found that E25 was detectable in the serum during aerosol treatment, although serum IgE did not change significantly in any of the three groups during treatment. In addition, both doses of E25 were no more effective than placebo in attenuating the early phase responses to allergen at both times during treatment. Although aerosolized E25 was generally well tolerated, one subject receiving aerosolized E25 10 mg daily was found to have serum IgG and IgA antibodies to E25. We conclude that aerosol administration of an anti-IgE monoclonal antibody does not inhibit the airway responses to inhaled allergen in allergic asthmatic subjects. We speculate that the observed lack of efficacy may be due to the inability of aerosol route of delivery to result in high enough concentrations of E25 in the tissue compartments surrounding IgE effector cells to neutralize IgE arising from local airway and pulmonary sources and IgE arising from the vascular space. Additionally, the aerosol route of delivery of monoclonal antibodies may be more immunogenic than the parenteral route. PMID- 10471636 TI - Effects of salmeterol on arterial blood gases in patients with stable chronic obstructive pulmonary disease. Comparison with albuterol and ipratropium. AB - Administration of beta-adrenergic agonist bronchodilators to patients with airways obstruction commonly results in transient decreases in Pa(O(2)) levels despite bronchodilation, an effect that has been attributed to these drugs' pulmonary vasodilator action. We compared the acute effects on gas exchange of salmeterol with those of albuterol and the anticholinergic agent ipratropium in 20 patients with stable chronic obstructive pulmonary disease (COPD). Each agent was given in recommended dosage on separate days in a double-blind, crossover format, and the patients' arterial blood gases (ABGs) were measured at baseline and at intervals to 120 min. Small but statistically significant declines in Pa(O(2)), the primary outcome variable, were found after administration of both salmeterol and albuterol. The decline in PaO2 after salmeterol was of lesser magnitude but was more prolonged than that after albuterol, the greatest mean change being -2.74 +/- 0.89 mm Hg (mean +/- SEM) at 30 min after salmeterol, and 3.45 +/- 0.92 mm Hg at 20 min after albuterol. Following ipratropium, the corresponding change was -1.32 +/- 0.85 mm Hg at 20 min. These declines, which were almost entirely attributable to increases in the alveolar-arterial difference in oxygen tension Delta(A-a)DO2 tended to be more marked in subjects with higher baseline PaO2 values. No subject experienced a decline in PaO2 to levels below 59 mm Hg. There were no significant differences among the three drugs studied. We conclude that despite small decreases in PaO2 after each of the three drugs, the declines were small transient, and of doubtful clinical significance. PMID- 10471637 TI - Nitration of proteins in bronchoalveolar lavage fluid from patients with acute respiratory distress syndrome receiving inhaled nitric oxide. AB - Inhaled nitric oxide (.NO) is used to improve gas exchange and reduce pulmonary vascular resistance (PVR) in patients with the acute respiratory distress syndrome (ARDS). Although controlled studies have shown no survival benefit, some investigators have suggested that inhaled.NO may have antiinflammatory properties under these circumstances. In contrast, others have speculated that.NO given by inhalation could be cytotoxic, as it combines with superoxide at near diffusion limited rates to produce the highly reactive oxidant peroxynitrite (ONOO(-)). We therefore quantified levels of 3-nitrotyrosine, a marker for ONOO(-) formation, in bronchoalveolar lavage fluid (BAL) from patients with ARDS receiving inhaled.NO, and from patients with comparable lung injury who were not so treated. We also measured levels of 3-chlorotyrosine as an index of neutrophil activation to assess indirectly the effects of inhaled.NO on lung inflammation. Patients receiving .NO had increased levels of 3-nitrotyrosine (6.76 +/- 2.79 versus 0.4 +/- 0.15 nmol/mg of protein, p < 0.05) and 3-chlorotyrosine (7.97 +/- 2.74 versus 1. 53 +/- 1.09 nmol/mg of protein, p < 0.05) in BAL protein compared with controls. In patients with ARDS, inhaled.NO increases the formation of 3 nitrotyrosine and is accompanied by an increase in levels of 3-chlorotyrosine (a marker of neutrophil activation). The possible long-term consequences of these observations remain to be evaluated. PMID- 10471638 TI - Airway remodeling and repair. PMID- 10471639 TI - The diagnostic approach to acute venous thromboembolism. Clinical practice guideline. American Thoracic Society. PMID- 10471640 TI - Development of a chemiluminescence competitive PCR for the detection and quantification of parvovirus B19 DNA using a microplate luminometer. AB - BACKGROUND: Quantitative PCR of viral nucleic acids can be useful clinically in diagnosis, risk assessment, and monitoring of antiviral therapy. We wished to develop a chemiluminescence competitive PCR (cPCR) for parvovirus B19. METHODS: Parvovirus DNA target sequences and competitor sequences were coamplified and directly labeled. Amplified products were then separately hybridized by specific biotin-labeled probes, captured onto streptavidin-coated ELISA microplates, and detected immunoenzymatically using chemiluminescent substrates of peroxidase. Chemiluminescent signals were quantitatively analyzed by a microplate luminometer and were correlated to the amounts of amplified products. RESULTS: Luminol-based systems displayed constant emission but had a higher detection limit (100-1000 genome copies) than the acridan-based system (20 genome copies). The detection limit of chemiluminescent substrates was lower (20 genome copies) than colorimetric substrates (50 genome copies). In chemiluminescence cPCR, the titration curves showed linear correlation above 100 target genome copies. Chemiluminescence cPCR was positive in six serum samples from patients with parvovirus infections and negative in six control sera. CONCLUSIONS: The chemiluminescence cPCR appears to be a sensitive and specific method for the quantitative detection of viral DNAs. PMID- 10471641 TI - Quantitative reverse transcription-PCR assay with an internal standard for the detection of prostate-specific antigen mRNA. AB - BACKGROUND: Circulating prostate cells can be detected with a reverse transcription-PCR (RT-PCR) assay for prostate-specific antigen (PSA) mRNA. We have developed a new quantitative RT-PCR method for measuring PSA mRNA. METHODS: The method uses a PSA-like internal standard (IS) mRNA that is added into the sample at the beginning of the RNA extraction and coamplified by RT-PCR with the PSA in the sample. After PCR amplification, the IS and PSA products are selectively detected by hybridization in a microtitration plate using probes labeled with fluorescent europium chelates. RESULTS: The method was validated with PSA and IS mRNAs and PSA-expressing cells to obtain a detection limit of 50 PSA mRNA copies (i.e., signal 2 times the mean of zero signal), linearity up to 10(6) copies, and detection of a single PSA-expressing cell. In preliminary evaluations, 60% (n = 10) of the prostate cancer patients with skeletal metastases gave results above the detection limit (500 PSA mRNA copies in 5 mL of blood). The total number of PSA copies ranged from 900 +/- 200 to 44 100 +/- 4900 (mean +/- SD) in the samples, corresponding to approximately 1-100 PSA-expressing cells in 5 mL of blood. In the controls (n = 34), none of the healthy females and 2 of 19 healthy males had detectable PSA mRNA [700 +/- 100 and 2000 +/- 900 (mean +/- SD) PSA mRNA copies in 5 mL of blood for the 2 males]. CONCLUSIONS: The assay provides sensitive and quantitative detection of PSA mRNA expression from blood samples and can be used to establish the clinically significant number of PSA mRNA copies in prostate cancer. PMID- 10471642 TI - Microheterogeneity of serum glycoproteins in patients with chronic alcohol abuse compared with carbohydrate-deficient glycoprotein syndrome type I. AB - BACKGROUND: Chronic alcohol abuse alters the normal N-glycosylation of transferrin, producing the carbohydrate-deficient transferrin isoforms. This alteration could be similar to that present in patients with carbohydrate deficient glycoprotein syndrome type 1 (CDG1). We thus compared the alterations of N-glycans present in patients with alcoholism and patients with CDG1. METHODS: The N-glycans of serum glycoproteins were compared in sera of patients with alcoholism, patients with CDG1, and controls by two-dimensional electrophoresis, neuraminidase, peptide:N-glycosidase F, and endoglycosidase F2 treatments. A specific antibody directed against the amino acid sequence surrounding the N-432 N-glycosylation site of transferrin was prepared (SZ-350 antibody). RESULTS: In patients with alcoholism, the abnormal transferrin and alpha(1)-antitrypsin isoforms were devoid of a variable number of entire N-glycan moieties and were identical with those present in CDG1. In the serum of patients with alcoholism, this finding was less pronounced than in CDG1. In contrast to CDG1, there was no decrease in clusterin or serum amyloid P in patients with alcoholism. The SZ-350 antibody recognized only transferrin isoforms with one or no N-glycan moieties. CONCLUSION: Antibodies directed against specific N-glycosylation sites of glycoproteins could be useful for developing more specific immunochemical tests for the diagnosis of chronic alcohol abuse. PMID- 10471643 TI - Standardization of creatine kinase-MB (CK-MB) mass assays: the use of recombinant CK-MB as a reference material. AB - BACKGROUND: The AACC assembled a committee to identify and validate a standard creatine kinase MB isoenzyme (CK-MB) material to improve the comparability of CK MB mass assays. METHODS: Three protocols were used. In protocol I, various CK-MB materials prepared in different matrices were screened as candidate standards. In protocol II, participating manufacturers calibrated their systems with concentrates of human heart CK-MB and then tested 20 patient samples to evaluate calibration bias. In protocol III, participating manufacturers calibrated their immunoassay systems using recombinant CK-MB2 (rCK-MB2) diluted into their respective sample diluents and measured 50 samples. RESULTS: Candidate materials showed high recovery in stripped human serum, but bias improved only from 59% to 38%. These data led to the use of human heart CK-MB diluted in each manufacturer's sample diluent. This strategy reduced bias from 31% to 15%. Because human heart CK-MB is difficult to provide, a lyophilized source of CK-MB2 was identified. rCK-MB2 was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, reversed-phase HPLC, intrinsic protein fluorescence, circular dichroism, agarose gel electrophoresis, immunoreactivity studies, high and low temperature stability, and reconstituted stability to be equivalent to human heart CK-MB. Calibration of immunoassay systems with rCK-MB2 added into each respective manufacturer's sample diluent showed a 13% between-manufacturer bias. CONCLUSION: Lyophilized rCK-MB2 was determined suitable for use as a reference material for CK-MB mass assays. PMID- 10471644 TI - Long-distance PCR-based screening for large rearrangements of the LDL receptor gene in Korean patients with familial hypercholesterolemia. AB - BACKGROUND: The LDL receptor is a cell-surface protein that regulates plasma cholesterol by specific uptake of LDL particles from the blood circulation. Familial hypercholesterolemia (FH) results from defective catabolism of LDL, which is caused by mutations in the LDL-receptor gene. METHODS: For the rapid and reliable detection of large rearrangements in the LDL-receptor gene, we established a screening method based on long-distance PCR as an alternative to Southern-blot hybridization. Using long-distance PCR, 45 unrelated Korean subjects heterozygous for FH were screened to assess the frequency and nature of major structural rearrangements in the LDL-receptor gene. RESULTS: Two different deletion mutations, FH6 (same type as FH3 and FH311) and FH 32, were detected in four families by long-distance PCR. Detailed restriction mapping and sequence analysis showed that FH6 was a 5.71-kb deletion extending from intron 8 to intron 12 and that FH32 was a 2-kb deletion extending from intron 6 to intron 7. Sequence analysis for the breakpoints of all deletions detected in Korean FH patients showed that only the left arms of the Alu repetitive sequences were involved in the deletion event. CONCLUSIONS: The screening method based on long distance PCR provides a powerful strategy for the detection of large rearrangements in the LDL-receptor gene and is a rapid and reliable screening alternative to Southern-blot hybridization. PMID- 10471645 TI - Characterization of apolipoprotein E-containing lipoproteins in cerebrospinal fluid: effect of phenotype on the distribution of apolipoprotein E. AB - BACKGROUND: Apolipoprotein (apo) E, one of the main apolipoproteins in the central nervous system, may play an important role in lipid metabolism; however, the details of its function are poorly understood. In this study, we characterized apoE-containing lipoproteins in cerebrospinal fluid (CSF) and examined the effect of apoE phenotype on the distribution of apoE among the lipoprotein fractions. METHODS: CSF lipoproteins were fractionated by gel filtration and ultracentrifugation, and then characterized by electrophoresis, immunoblot, electron microscopy, and analysis of apoE, total cholesterol, and phospholipid concentrations. RESULTS: The ratio of sialylated to nonsialylated apoE was higher in CSF than in serum. However, the fundamental forms containing apoE homodimers or heterodimers [such as apo(E-AII) and apo(AII-E2-AII) complexes] were similar in CSF and serum. apoE-containing lipoproteins were fractionated at densities of <1.006, 1.063-1.125, and 1.125-1.21 kg/L. Neither apoE nor apoAI was detected in the fraction with a density range of 1.006-1.063 kg/L. The diameters of the lipoprotein particles with densities of <1.006, 1.063 1.125, and 1.125-1.21 kg/L were 16.7 +/- 3.1, 14.0 +/- 3.2, and 11.6 +/- 2.8 nm (mean +/- SD, n = 200), respectively. All of these lipoproteins exhibited a spherical structure. The distribution profile of apoE-containing lipoproteins was affected by the apoE phenotype. A relatively large amount of apoE-containing lipoproteins was isolated from the fraction with a density >1.125 kg/L obtained from CSF associated with apoE2 or apoE3. This tendency was more obvious in CSF associated with apoE2 than in CSF without apoE2. apoE-containing lipoproteins were predominantly observed in the fraction with a density of <1.006 kg/L obtained from CSF associated with apoE4. CONCLUSIONS: The lipoproteins in CSF have a unique composition that is different from that of the lipoproteins in plasma. However, the differences in diameter between the CSF fractions were not as large as for the serum fractions. Our data suggest that the apoE phenotype may affect the distribution profile of apoE-containing lipoproteins in the CSF. This would mean that the metabolism of apoE-containing lipoproteins depends on the apoE isoform present. PMID- 10471646 TI - Increased ability of LDL from normolipidemic type 2 diabetic women to generate peroxides. AB - BACKGROUND: We assessed the ability of LDL from 30 type 1 diabetic patients (18 men, 12 women), 65 type 2 diabetic patients (35 men, 30 women), and 35 controls (19 men, 16 women) to generate peroxides. The men and women in the diabetic groups were studied separately and matched for age, body mass index, duration of diabetes, glycohemoglobin, and conventional lipid characteristics according to the presence or absence of hyperlipidemia. METHODS: The ability of LDL to form peroxides was assessed by measuring the thiobarbituric acid-reactive substances corrected for LDL-cholesterol [ratio of malondialdehyde (MDA) to LDL cholesterol]. LDL particle size was expressed as the ratio of LDL-cholesterol to apolipoprotein B (LDL-cholesterol/apoB). RESULTS: The MDA/LDL-cholesterol ratio was higher in type 1 and type 2 diabetic patients with hyperlipidemia than in controls. The MDA/LDL-cholesterol ratio was also higher in type 2 normolipidemic women than in controls (P <0.01). The LDL-cholesterol/apoB ratio was lower in type 2 diabetic women than in type 2 diabetic men (P <0.05). The MDA/LDL cholesterol ratio was negatively correlated with the LDL-cholesterol/apoB ratio (r = -0.78, P <0.001) in hyperlipidemic type 1 (not type 2) diabetic patients. In normolipidemic type 2 diabetic patients, the MDA/LDL-cholesterol ratio was also negatively correlated with the LDL-cholesterol/apoB ratio (r = -0.75, P <0.001) because of the highly significant negative correlation in type 2 diabetic women (r = -0.89, P <0.01). CONCLUSIONS: LDL from well-controlled type 2 diabetic women is smaller and more prone to form peroxides. This could explain why diabetic women are at greater risk of cardiovascular disease. PMID- 10471647 TI - Analytical validation of the PRO-Trac II ELISA for the determination of tacrolimus (FK506) in whole blood. AB - BACKGROUND: The analytical validation of multiple lots of the PRO-Trac II ELISA (DiaSorin) for the determination of tacrolimus in whole blood is described. METHODS: The analytical parameters assessed included analytical sensitivity, dilution linearity, functional sensitivity, values in samples containing no tacrolimus, intra- and interassay precision, supplementation and recovery, metabolite cross-reactivity, interference studies, and method comparisons HPLC tandem mass spectrometry (HPLC/MS/MS) and the IMx Tacrolimus II multiparticle enzyme immunoassay. Where appropriate, assessments were performed according to NCCLS guidelines. RESULTS: The mean analytical detection limit was <0.25 microg/L for all lots, whereas the functional sensitivity was 1.0 microg/L. Excellent linear correlation (r = 0.985) was observed for dilution linearity. The intraassay imprecision was <7%, and the total imprecision by ANOVA was <10%. Recovery was 109% +/- 11%. Metabolite cross-reactivity was consistent with previous reports for this antibody. No interference was observed for 35 tested drugs. Method comparison with HPLC/MS/MS showed no statistically significant differences. Samples exhibited stability through four freeze/thaw cycles and for 1 week at room temperature. CONCLUSION: These data demonstrate that the PRO-Trac II ELISA is a robust, accurate, and precise tool for the assessment and management of tacrolimus blood concentrations in transplant patients. PMID- 10471648 TI - Simple method for determination of thiocyanate in urine. AB - BACKGROUND: It would be useful to develop a simple kit method for determination of thiocyanate in urine, which could be used to monitor cyanide overload in cassava-consuming populations. METHODS: The method was based on the quantitative oxidation of thiocyanate in acid permanganate at room temperature in a closed vial with liberation of HCN, which reacted with a picrate paper. For semiquantitative analysis in the field, the colored picrate paper was matched with a color chart prepared using known amounts of KSCN. In the laboratory, a more accurate result was obtained by elution of the colored complex in water and measurement of the absorbance at 510 nm. Over the range 0-100 mg/L, there was a linear relationship given by the equation: thiocyanate content (mg/L) = 78 x absorbance. RESULTS: The picrate thiocyanate method gave no interference with urine samples containing protein at less than 7 g/L, 21 amino acids, histamine, glucose, NaCl, urea, blood, and linamarin. For 53 urine samples analyzed by an accurate column method and the thiocyanate picrate method, a regression line gave very good agreement (r(2) = 1. 000). Quantitative recoveries of thiocyanate added to urine samples were obtained with the picrate method. CONCLUSIONS: A simple picrate kit for determination of thiocyanate in urine was developed and is available free of charge for workers in developing countries. PMID- 10471649 TI - Drug monitoring of antiretroviral therapy for HIV-1 infection: method validation and results of a pilot study. AB - BACKGROUND: Antiretroviral therapy for HIV-1 infection has become increasingly complex. The availability of new and potent drugs and progress in understanding the pathogenesis of HIV-1 infection have led to the establishment of new treatment paradigms. The varying dosing regimens, associated toxicities, and the potential for drug-drug and food-drug interactions further complicate treatment. This complexity contributes to patient nonadherence. Because clinicians have no tools to monitor adherence or drug-drug interactions and because response requires that therapy exceed the known inhibiting concentration, serum monitoring of antiretroviral therapy may play a role in improving treatment of HIV-1 infection. We report methods to quantify serum concentrations of antiretroviral drugs used to treat HIV-1 infection, precision and interference studies of these methods, and results observed in a pilot evaluation of blood serum concentrations from 12 human subjects. METHODS: HPLC offers adequate sensitivity to measure peak or trough serum concentrations of delavirdine, lamivudine, nevirapine, indinavir, nelfinavir, ritonavir, and saquinavir and peak serum concentrations of stavudine, zidovudine, and didanosine with reasonable precision. RESULTS: Peak indinavir serum concentrations in most patients were in the range of 1-10 mg/L, and trough concentrations were in the range of 0.1-0.5 mg/L. Peak stavudine concentrations were in the range of 0.3-1.3 mg/L, and trough concentrations were in the range of 0.1-0.5 mg/L. Peak zidovudine concentrations were in the range of 0.1-1.1 mg/L. CONCLUSIONS: Because this was a blood serum concentration-seeking pilot study to evaluate analytic performance, we do not report on the correlation of drug response to blood concentration. However, the concentrations observed in patients are generally consistent with blood concentrations reported from studies of monotherapy. PMID- 10471650 TI - Inhibition of stimulated interleukin-2 production in whole blood: a practical measure of cyclosporine effect. AB - BACKGROUND: Prediction of cyclosporine (CSA) efficacy and toxicity in individual patients is difficult. There is no practical, biologically relevant, pharmacodynamic measure of CSA effect. A major effect of CSA is to decrease interleukin-2 (IL-2) production; however, measurement of this effect in isolated lymphocytes as a marker of response to CSA has been problematic. METHODS: CSA inhibition of phytohemagglutinin-P (PHA)-stimulated IL-2 production, measured by ELISA, was studied ex vivo in whole blood drawn before, and after subjects received 4 mg/kg oral CSA. RESULTS: Four hours after CSA was administered, the mean (+/- SD) CSA concentration was 702 +/- 196 microg/L and PHA-stimulated IL-2 production decreased by 68.7% +/- 17.2% (P <0.0001; n = 17). Twenty-four hours after CSA was administered, concentrations were low (64 +/- 24 microg/L), with no inhibition of IL-2 production. A rapid, concentration-dependent response occurred. Maximum CSA concentrations (944 +/- 187 microg/L) and maximum inhibition of IL-2 production (86.9% +/- 13.7%) occurred 90 min after subjects received CSA. In vitro, 32.5-1200 microg/L CSA also inhibited PHA-stimulated IL-2 production in whole blood in a dose-dependent fashion with a similar IC(50) (approximately 300-400 microg/L) ex vivo and in vitro. CONCLUSION: In the search for a pharmacodynamic marker to better guide immunosuppressive therapy, the relationship between this simple, biologically relevant measure of CSA effect and clinical outcome should be determined. PMID- 10471651 TI - Codeine concentration in hair after oral administration is dependent on melanin content. AB - BACKGROUND: Analysis of drugs in hair has been used on a qualitative basis to estimate earlier exposure to drugs. Clinical applications are rare because of the lack of dose-response relationships in the studies performed to date, and questions remain regarding the mechanisms of drug incorporation into hair. Several human studies have shown differences in drug accumulation between pigmented and nonpigmented hair. However, the melanin concentration in hair was not determined and correlated to the amount of drug incorporated. METHODS: Nine human subjects were given codeine as a single oral dose, and plasma codeine concentrations were determined for 24 h, using gas chromatography-mass spectrometry. Hair samples were obtained weekly for a month. Total melanin, eumelanin, and codeine were measured quantitatively in hair samples by spectrophotometry, HPLC, and gas chromatography-mass spectrometry, respectively. RESULTS: There was an exponential relationship between codeine and melanin concentrations in hair, (r(2) = 0.95 with total melanin and r(2) = 0.83 with eumelanin). After normalizing the results by the area under the curve for codeine in plasma, we obtained r(2) = 0.86 for codeine vs total melanin and r(2) = 0.90 vs eumelanin. CONCLUSIONS: Our results stress the importance of melanin determination when measuring drugs in hair. We postulate that analysis of drug concentration in hair may be worthwhile in the monitoring of drug compliance if the results are normalized for melanin content. PMID- 10471652 TI - Predictive value of cord blood hematological indices and hemoglobin Barts for the detection of heterozygous alpha-thalassemia-2 in an African-Caribbean population. AB - BACKGROUND: Cord blood hemoglobin Barts (HbBarts) and hemocytometric indices may be used for classification of newborns into those without alpha-thalassemia-2 (alphaalpha/alphaalpha) and with heterozygous alpha-thalassemia-2 ( alpha(3.7)/alphaalpha). We investigated by logistic regression analysis whether the combination of HbBarts and hemocytometric indices improves classification compared with classification based on a single analyte. METHODS: HbBarts percentages and hemocytometric indices were determined in cord blood of 208 consecutive newborns in Curacao (Netherlands Antilles). Of these, 157 had alphaalpha/alphaalpha and 51 had -alpha(3.7)/alphaalpha, as established by DNA analysis. RESULTS: Between-group differences were significant for erythrocytes, mean cell volume, mean cell hemoglobin (MCH), mean cell hemoglobin concentration, platelets, hemoglobin F(0) (HbF(0)), and HbBarts. The Logit equation of the logistic regression model, using MCH (pg) and HbBarts (%), was: 42.7164 + 5.7916(HbBarts) - 1.3110(MCH). A sensitivity of 100% was reached at a Logit value of -3.70. The corresponding specificity was 62.2%, and the predictive value of a positive test (PV+) was 46.3% (95% confidence interval, 37.0-55.7%). The relative information gains were as follows: 88% for the HbBarts-MCH combination, 26% for MCH (not significant), and 0% for HbBarts compared with the 24.6% alpha(3.7)/alphaalpha prevalence. CONCLUSION: Combined use of cord blood HbBarts and MCH improves classification compared with classification based on single hemocytometric indices. PMID- 10471654 TI - Total and free deoxypyridinoline after acute osteoclast activity inhibition. AB - BACKGROUND: Deoxypyridinoline (Dpd) is one of the two pyridinium cross-links that provide structural rigidity to type I collagen in bone. During osteoclastic resorption, Dpd is released into circulation and is excreted in the urine in free and peptide-bound forms. Free and total Dpd are highly correlated, but whether the free-to-total cross-link ratio is constant in both normal and high bone turnover states remains controversial. To compare free and total Dpd performance in a physiological condition, urinary free and total Dpd were measured after a short-term inhibition of osteoclast activity such as that induced by an oral calcium load. METHODS: Total and free Dpd were measured by HPLC and by immunosorbent assay, respectively, in two groups of subjects, one (calcium treated; n = 16) taking calcium and the other not (control; n = 9). RESULTS: The urinary excretion of total Dpd at 2 and 4 h after oral calcium loading was decreased compared with controls. By contrast, changes in free Dpd were similar in the calcium-treated and control groups, reflecting only circadian rhythm. CONCLUSIONS: Total and free Dpd do not show comparable sensitivity in detecting short-term inhibition of osteoclast activity. The degradation process of peptide bound to free Dpd could render free Dpd insensitive to acute changes of osteoclast activity. PMID- 10471653 TI - A collection method and high-sensitivity enzyme immunoassay for sweat pyridinoline and deoxypyridinoline cross-links. AB - BACKGROUND: Collagen cross-link molecules such as pyridinoline (PYD), deoxypyridinoline (DPD), and N-terminal cross-linked peptides (NTX) have been measured in urine as indices of bone resorption. However, very little is known regarding the excretion of pyridinolines into other biological fluids. We report a collection device, normalizing analyte, and high-sensitivity immunoassay for quantitative analysis of free pyridinoline cross-links in sweat. METHODS: Flame atomic emission and ion-selective electrode techniques were used to measure potassium as a sweat volume marker. The Pyrilinks immunoassay for urine free pyridinolines was optimized to increase sensitivity for measurements in sweat. The precision, accuracy, and detection limit of this assay were characterized. To assess values and variability of sweat pyridinolines in human subjects, a nonocclusive skin patch was used to collect sweat samples from a reference group and from a mixed group experiencing accelerated bone resorption, postmenopausal women and men receiving gonadotropin-releasing hormone for prostate cancer. RESULTS: The immunoassay intra- and interassay variations were 10 in 90% of the samples) but also isolated these NRBCs in 78 consecutive maternal samples. CONCLUSION: Antibody 2B7.4 shows promise for the isolation of NRBCs from maternal blood and should allow studies concerning the source of these cells, fetal vs maternal, and the factors controlling their prevalence. PMID- 10471677 TI - Evaluation of measurement sites for noninvasive blood glucose sensing with near infrared transmission spectroscopy. AB - Six putative measurement sites were evaluated for noninvasive sensing of blood glucose by first-overtone near-infrared spectroscopy. The cheek, lower lip, upper lip, nasal septum, tongue, and webbing tissue between the thumb and forefinger were examined. These sites were evaluated on the basis of their chemical and physical properties as they pertain to the noninvasive measurement of glucose. Critical features included the effective optical pathlength of aqueous material within the tissue and the percentage of body fat within the optical path. Aqueous optical paths of 5 mm are required to measure clinically relevant concentrations of glucose in the first-overtone region. All of the tested sites met this requirement. The percentage of body fat affects the signal-to-noise ratio of the measurement and must be minimized for reliable glucose sensing. The webbing tissue contains a considerable amount of fat tissue and is clearly the worse measurement site. All other sites possess substantially less fat, with the least amount of fat in tongue tissue. For this reason, the tongue provides spectra with the highest signal-to-noise ratio and is, therefore, the site of choice on the basis of spectral quality. PMID- 10471678 TI - Aptamers: an emerging class of molecules that rival antibodies in diagnostics. AB - Antibodies, the most popular class of molecules providing molecular recognition needs for a wide range of applications, have been around for more than three decades. As a result, antibodies have made substantial contributions toward the advancement of diagnostic assays and have become indispensable in most diagnostic tests that are used routinely in clinics today. The development of the systematic evolution of ligands by exponential enrichment (SELEX) process, however, made possible the isolation of oligonucleotide sequences with the capacity to recognize virtually any class of target molecules with high affinity and specificity. These oligonucleotide sequences, referred to as "aptamers", are beginning to emerge as a class of molecules that rival antibodies in both therapeutic and diagnostic applications. Aptamers are different from antibodies, yet they mimic properties of antibodies in a variety of diagnostic formats. The demand for diagnostic assays to assist in the management of existing and emerging diseases is increasing, and aptamers could potentially fulfill molecular recognition needs in those assays. Compared with the bellwether antibody technology, aptamer research is still in its infancy, but it is progressing at a fast pace. The potential of aptamers may be realized in the near future in the form of aptamer-based diagnostic products in the market. In such products, aptamers may play a key role either in conjunction with, or in place of, antibodies. It is also likely that existing diagnostic formats may change according to the need to better harness the unique properties of aptamers. PMID- 10471679 TI - Noninvasive prediction of glucose by near-infrared diffuse reflectance spectroscopy. AB - BACKGROUND: Self-monitoring of blood glucose by diabetics is crucial in the reduction of complications related to diabetes. Current monitoring techniques are invasive and painful, and discourage regular use. The aim of this study was to demonstrate the use of near-infrared (NIR) diffuse reflectance over the 1050-2450 nm wavelength range for noninvasive monitoring of blood glucose. METHODS: Two approaches were used to develop calibration models for predicting the concentration of blood glucose. In the first approach, seven diabetic subjects were studied over a 35-day period with random collection of NIR spectra. Corresponding blood samples were collected for analyte analysis during the collection of each NIR spectrum. The second approach involved three nondiabetic subjects and the use of oral glucose tolerance tests (OGTTs) over multiple days to cause fluctuations in blood glucose concentrations. Twenty NIR spectra were collected over the 3.5-h test, with 16 corresponding blood specimens taken for analyte analysis. RESULTS: Statistically valid calibration models were developed on three of the seven diabetic subjects. The mean standard error of prediction through cross-validation was 1.41 mmol/L (25 mg/dL). The results from the OGTT testing of three nondiabetic subjects yielded a mean standard error of calibration of 1.1 mmol/L (20 mg/dL). Validation of the calibration model with an independent test set produced a mean standard error of prediction equivalent to 1.03 mmol/L (19 mg/dL). CONCLUSIONS: These data provide preliminary evidence and allow cautious optimism that NIR diffuse reflectance spectroscopy using the 1050 2450 nm wavelength range can be used to predict blood glucose concentrations noninvasively. Substantial research is still required to validate whether this technology is a viable tool for long-term home diagnostic use by diabetics. PMID- 10471680 TI - Thin film biosensor for rapid visual detection of nucleic acid targets. AB - BACKGROUND: We have developed a silicon-based biosensor that generates a visual signal in response to nucleic acid targets. METHODS: In this system, capture oligonucleotide probes are immobilized on the surface of the biosensor. Interaction of the capture probes with a complementary target and a biotinylated detector oligonucleotide allows initiation of formation of an organic thin film on the biosensor. Thin film formation is completed by enzymatic activity of peroxidase conjugated to an anti-biotin antibody. Peroxidase catalyzes deposition of an insoluble product onto the silicon surface, generating a uniform thin film. The increased thickness on the surface alters the perceived color of the biosensor through changes in the interference patterns of reflected light from the surface, causing a color change from gold to purple. RESULTS: The biosensor results may be evaluated by direct visual inspection or quantified by ellipsometry. Results are obtained in 25 min with a detection limit of 5 pmol/L (150 amol/sample). Selectivity of the biosensor is demonstrated by discrimination of single nucleotide mismatches. Multitarget arrays are also analyzed with the thin film biosensor, and the system is capable of detecting targets from human serum and urine. CONCLUSIONS: The biosensor surface is inexpensive to produce, and the assay format is simple and rapid. The thin film biosensor is adaptable to a wide variety of nucleic acid detection applications, including rapid diagnostic testing for infectious disease panels, antibiotic resistance panels, or allelic discrimination of specific genetic markers. PMID- 10471681 TI - Analytical characterization of electrochemical biosensor test strips for measurement of glucose in low-volume interstitial fluid samples. AB - BACKGROUND: Minimally invasive interstitial fluid (ISF) sampling and glucose measurement technologies were integrated into a hand-held device for diabetic glucose monitoring investigations. METHODS: Conventional electrochemical test strip technology (Bayer Glucometer Elite) was adapted to measure glucose in small (0.5-2.0 microL) samples of ISF. Test strip glucose measurements were performed on a commercial potentiostat and were compared to various reference glucose methodologies (YSI 2300 analyzer, microhexokinase procedure, Bayer Glucometer Elite). Characterizations of the integrated ISF sampling-glucose test strip design included accuracy and precision in various sample media (saline, ISF surrogates, diabetic ISF samples), sample volume dependence, test strip sterilization studies (electron beam, gamma irradiation), and diabetic ISF sampling and glucose measurements. RESULTS: Glucose measurements were free from significant media effects. Sample volume variations (0.6-3.2 microL) revealed only modest dependence of glucose measurement bias on sample volume (-1.5% per microliter). Sterilization treatments had only a minor impact on glucose response and test strip aging and no significant impact on interferent responses of the glucose test strips. Diabetic subject testing under minimum fasting conditions of at least 2 h with integrated ISF sampling and glucose measurement gave low ISF glucose measurement imprecision (CV, 4%) and mean glucose results that were indistinguishable from reference (microhexokinase) ISF glucose measurements and from capillary blood glucose measurements (Glucometer Elite). CONCLUSIONS: Conventional single-use, electrochemical glucose test strip and ISF collection technologies can be readily integrated to provide real-time ISF sampling and glucose measurements for diabetic monitoring applications. PMID- 10471682 TI - Site-to-Site Variation of Glucose in Interstitial Fluid Samples and Correlation to Venous Plasma Glucose. PMID- 10471683 TI - Immobilization of Monolayers of Fc-binding Receptors on Planar Solid Supports. PMID- 10471684 TI - RAMP(TM): A Rapid, Quantitative Whole Blood Immunochromatographic Platform for Point-of-Care Testing. PMID- 10471685 TI - Highly Miniaturized and Integrated Biosensor for Analysis of Whole Blood Samples. PMID- 10471686 TI - Management of Interferences in a Transdermal, Noninvasive Glucose Monitoring Device. PMID- 10471687 TI - Design of a Biosensor for Continual, Transdermal Glucose Monitoring. PMID- 10471688 TI - Development of Kinetic Ligand-binding Assays Using a Fiber Optic Sensor. PMID- 10471690 TI - Lateral Tissue Inhomogeneity: A Missing Link in Photoplethysmographic Noninvasive Measurement of Arterial Blood Constituents. PMID- 10471689 TI - New Miniaturized Highly Sensitive Immunoassay Device for Quantitative Measurement of Soluble or Particular Antigen or Antibodies in a Liquid Sample. PMID- 10471691 TI - Materials for Fabricating Biosensors for Transdermal Glucose Monitoring. PMID- 10471692 TI - Multichannel Electrochemical Detection System for Quantitative Monitoring of PCR Amplification. PMID- 10471693 TI - Simultaneous Quantification of Six Human Cytokines in a Single Sample Using Microparticle-based Flow Cytometric Technology. PMID- 10471694 TI - J. B. S. Haldane (1949) on infectious disease and evolution. PMID- 10471695 TI - Mutational adaptation of Escherichia coli to glucose limitation involves distinct evolutionary pathways in aerobic and oxygen-limited environments. AB - Mutational adaptations leading to improved glucose transport were followed with Escherichia coli K-12 growing in glucose-limited continuous cultures. When populations were oxygen limited as well as glucose limited, all bacteria within 280 generations contained mutations in a single codon of the ptsG gene. V12F and V12G replacements in the enzyme IIBC(Glc) component of the glucose phosphotransferase system were responsible for improved transport. In stark contrast, ptsG mutations were uncommon in fully aerobic glucose-limited cultures, in which polygenic mutations in mgl, mlc, and malT (regulating an alternate high affinity Mgl/LamB uptake pathway) spread through the adapted population. Hence the same organism adapted to the same selection (glucose limitation) by different evolutionary pathways depending on a secondary environmental factor. The clonal diversity in the adapted populations was also significantly different. The PtsG V12F substitution under O(2) limitation contributed to a universal "winner clone" whereas polygenic, multiallelic changes led to considerable polymorphism in aerobic cultures. Why the difference in adaptive outcomes? E. coli physiology prevented scavenging by the LamB/Mgl system under O(2) limitation; hence, ptsG mutations provided the only adaptive pathway. But ptsG mutations in aerobic cultures are overtaken by mgl, mlc, and malT adaptations with better glucose scavenging ability. Indeed, when an mglA::Tn10 mutant with an inactivated Mgl/LamB pathway was introduced into two independent aerobic chemostats, adaptation of the Mgl(-) strain involved the identical ptsG mutation found under O(2)-limited conditions with wild-type or Mgl(-) bacteria. PMID- 10471696 TI - Identification of SAS4 and SAS5, two genes that regulate silencing in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, chromatin-mediated silencing inactivates transcription of the genes at the HML and HMR cryptic mating-type loci and genes near telomeres. Mutations in the Rap1p and Abf1p binding sites of the HMR-E silencer (HMRa-e**) result in a loss of silencing at HMR. We characterized a collection of 15 mutations that restore the alpha-mating phenotype to MATalpha HMRa-e** strains. These mutations defined three complementation groups, two new groups and one group that corresponded to the previously identified SAS2 gene. We cloned the genes that complemented members of the new groups and identified two previously uncharacterized genes, which we named SAS4 and SAS5. Neither SAS4 nor SAS5 was required for viability. Null alleles of SAS4 and SAS5 restored SIR4 dependent silencing at HMR, establishing that each is a regulator of silencing. Null alleles of SAS4 and SAS5 bypassed the role of the Abf1p binding site of the HMR-E silencer but not the role of the ACS or Rap1p binding site. Previous analysis indicated that SAS2 is homologous to a human gene that is a site of recurring translocations involved in acute myeloid leukemia. Similarly, SAS5 is a member of a gene family that included two human genes that are the sites of recurring translocations involved in acute myeloid leukemia. PMID- 10471697 TI - SAS4 and SAS5 are locus-specific regulators of silencing in Saccharomyces cerevisiae. AB - Sir2p, Sir3p, Sir4p, and the core histones form a repressive chromatin structure that silences transcription in the regions near telomeres and at the HML and HMR cryptic mating-type loci in Saccharomyces cerevisiae. Null alleles of SAS4 and SAS5 suppress silencing defects at HMR; therefore, SAS4 and SAS5 are negative regulators of silencing at HMR. This study revealed that SAS4 and SAS5 contribute to silencing at HML and the telomeres, indicating that SAS4 and SAS5 are positive regulators of silencing at these loci. These paradoxical locus-specific phenotypes are shared with null alleles of SAS2 and are unique among phenotypes of mutations in other known regulators of silencing. This work also determined that these SAS genes play roles that are redundant with SIR1 at HML, yet distinct from SIR1 at HMR. Furthermore, these SAS genes are not redundant with each other in silencing HML. Collectively, these data suggest that SAS2, SAS4, and SAS5 constitute a novel class of regulators of silencing and reveal fundamental differences in the regulation of silencing at HML and HMR. We provide evidence for a model that accounts for the observation that these SAS genes are both positive and negative regulators of silencing. PMID- 10471698 TI - Temperature-sensitive mutations in the Saccharomyces cerevisiae MRT4, GRC5, SLA2 and THS1 genes result in defects in mRNA turnover. AB - In a screen for factors involved in mRNA turnover, four temperature-sensitive yeast strains (ts1189, ts942, ts817, and ts1100) exhibited defects in the decay of several mRNAs. Complementation of the growth and mRNA decay defects, and genetic experiments, revealed that ts1189 is mutated in the previously unknown MRT4 gene, ts942 is mutated in GRC5 (encoding the L9 ribosomal protein), ts817 contains a mutation in SLA2 (encoding a membrane protein), and ts1100 contains a mutation in THS1 (encoding the threonyl-tRNA synthetase). Three of the four mutants (mrt4, grc5, and sla2) were not defective in protein synthesis, suggesting that these strains contain mutations in factors that may play a specific role in mRNA decay. The mRNA stabilization observed in the ths1 strain, however, could be due to the significant drop in translation observed in this mutant at 37 degrees. While the three interesting mutants appear to encode novel mRNA decay factors, at least one could be linked to a previously characterized mRNA decay pathway. The growth and mRNA decay defects of ts942 (grc5) cells were suppressed by overexpression of the NMD3 gene, encoding a protein shown to participate in a two-hybrid interaction with the nonsense-mediated decay protein Upf1p. PMID- 10471699 TI - Centromere mapping functions for aneuploid meiotic products: Analysis of rec8, rec10 and rec11 mutants of the fission yeast Schizosaccharomyces pombe. AB - Recent evidence suggests that the position of reciprocal recombination events (crossovers) is important for the segregation of homologous chromosomes during meiosis I and sister chromatids during meiosis II. We developed genetic mapping functions that permit the simultaneous analysis of centromere-proximal crossover recombination and the type of segregation error leading to aneuploidy. The mapping functions were tested in a study of the rec8, rec10, and rec11 mutants of fission yeast. In each mutant we monitored each of the three chromosome pairs. Between 38 and 100% of the chromosome segregation errors in the rec8 mutants were due to meiosis I nondisjunction of homologous chromosomes. The remaining segregation errors were likely the result of precocious separation of sister chromatids, a previously described defect in the rec8 mutants. Between 47 and 100% of segregation errors in the rec10 and rec11 mutants were due to nondisjunction of sister chromatids during meiosis II. In addition, centromere proximal recombination was reduced as much as 14-fold or more on chromosomes that had experienced nondisjunction. These results demonstrate the utility of the new mapping functions and support models in which sister chromatid cohesion and crossover position are important determinants for proper chromosome segregation in each meiotic division. PMID- 10471700 TI - Meiotic chromosome dynamics dependent upon the rec8(+), rec10(+) and rec11(+) genes of the fission yeast Schizosaccharomyces pombe. AB - During meiosis homologous chromosomes replicate once, pair, experience recombination, and undergo two rounds of segregation to produce haploid meiotic products. The rec8(+), rec10(+), and rec11(+) genes of the fission yeast Schizosaccharomyces pombe exhibit similar specificities for meiotic recombination and rec8(+) is required for sister chromatid cohesion and homolog pairing. We applied cytological and genetic approaches to identify potential genetic interactions and to gauge the fidelity of meiotic chromosome segregation in the mutants. The rec8(+) gene was epistatic to rec10(+) and to rec11(+), but there was no clear epistatic relationship between rec10(+) and rec11(+). Reciprocal (crossover) recombination in the central regions of all three chromosomes was compromised in the rec mutants, but recombination near the telomeres was nearly normal. Each of the mutants also exhibited a high rate of aberrant segregation for all three chromosomes. The rec8 mutations affected mainly meiosis I segregation. Remarkably, the rec10 and rec11 mutations, which compromised recombination during meiosis I, affected mainly meiosis II segregation. We propose that these genes encode regulators or components of a "meiotic chromatid cohesion" pathway involved in establishing, maintaining, and appropriately releasing meiotic interactions between chromosomes. A model of synergistic interactions between sister chromatid cohesion and crossover position suggests how crossovers and cohesion help ensure the proper segregation of chromosomes in each of the meiotic divisions. PMID- 10471701 TI - Pleiotropic defects caused by loss of the proteasome-interacting factors Rad23 and Rpn10 of Saccharomyces cerevisiae. AB - Rad23 is a member of a novel class of proteins that contain unprocessed ubiquitin like (UbL) domains. We showed recently that a small fraction of Rad23 can form an interaction with the 26S proteasome. Similarly, a small fraction of Rpn10 is a component of the proteasome. Rpn10 can bind multiubiquitin chains in vitro, but genetic studies have not clarified its role in vivo. We report here that the loss of both Rad23 and Rpn10 results in pleiotropic defects that are not observed in either single mutant. rad23Delta rpn10Delta displays slow growth, cold sensitivity, and a pronounced G2/M phase delay, implicating overlapping roles for Rad23 and Rpn10. Although rad23Delta rpn10Delta displays similar sensitivity to DNA damage as a rad23Delta single mutant, deletion of RAD23 in rpn10Delta significantly increased sensitivity to canavanine, a phenotype associated with an rpn10Delta single mutant. A mutant Rad23 that is unable to bind the proteasome ((DeltaUbL)rad23) does not suppress the canavanine or cold-sensitive defects of rad23Delta rpn10Delta, demonstrating that Rad23/proteasome interaction is related to these effects. Finally, the accumulation of multiubiquitinated proteins and the stabilization of a specific proteolytic substrate in rad23Delta rpn10Delta suggest that proteasome function is altered. PMID- 10471703 TI - The TamA protein fused to a DNA-binding domain can recruit AreA, the major nitrogen regulatory protein, to activate gene expression in Aspergillus nidulans. AB - The areA gene of Aspergillus nidulans encodes a GATA zinc finger transcription factor that activates the expression of a large number of genes subject to nitrogen metabolite repression. The amount and activity of the AreA protein under different nitrogen conditions is modulated by transcriptional, post transcriptional, and post-translational controls. One of these controls of AreA activity has been proposed to involve the NmrA protein interacting with the DNA binding domain and the extreme C terminus of AreA to inhibit DNA binding under nitrogen sufficient conditions. In contrast, mutational evidence suggests that the tamA gene has a positive role together with areA in regulating the expression of genes subject to nitrogen metabolite repression. This gene was identified by the selection of mutants resistant to toxic nitrogen source analogues, and a number of nitrogen metabolic activities have been shown to be reduced in these mutants. To investigate the role of this gene we have used constructs encoding the TamA protein fused to the DNA-binding domain of either the FacB or the AmdR regulatory proteins. These hybrid proteins have been shown to activate expression of the genes of acetate or GABA utilization, respectively, as well as the amdS gene. Strong activation was shown to require the AreA protein but was not dependent on AreA binding to DNA. The homologous areA gene of A. oryzae and nit-2 gene of Neurospora crassa can substitute for A. nidulans areA in this interaction. We have shown that the same C-terminal region of AreA and NIT-2 that is involved in the interaction with NmrA is required for the TamA-AreA interaction. However, it is unlikely that TamA requires the same residues as NmrA within the GATA DNA-binding domain of AreA. PMID- 10471702 TI - Genetic study of interactions between the cytoskeletal assembly protein sla1 and prion-forming domain of the release factor Sup35 (eRF3) in Saccharomyces cerevisiae. AB - Striking similarities between cytoskeletal assembly and the "nucleated polymerization" model of prion propagation suggest that similar or overlapping sets of proteins may assist in both processes. We show that the C-terminal domain of the yeast cytoskeletal assembly protein Sla1 (Sla1C) specifically interacts with the N-terminal prion-forming domain (Sup35N) of the yeast release factor Sup35 (eRF3) in the two-hybrid system. Sla1C and several other Sup35N-interacting proteins also exhibit two-hybrid interactions with the poly-Gln-expanded N proximal fragment of human huntingtin, which promotes Huntington disease associated aggregation. The Sup35N-Sla1C interaction is inhibited by Sup35N alterations that make Sup35 unable to propagate the [PSI(+)] state and by the absence of the chaperone protein Hsp104, which is essential for [PSI] propagation. In a Sla1(-) background, [PSI] curing by dimethylsulfoxide or excess Hsp104 is increased, while translational readthrough and de novo [PSI] formation induced by excess Sup35 or Sup35N are decreased. These data show that, in agreement with the proposed function of Sla1 during cytoskeletal formation, Sla1 assists in [PSI] formation and propagation, but is not required for these processes. Sla1(-) strains are sensitive to some translational inhibitors, and some sup35 mutants, obtained in a Sla1(-) background, are sensitive to Sla1, suggesting that the interaction between Sla1 and Sup35 proteins may play a role in the normal function of the translational apparatus. We hypothesize that Sup35N is involved in regulatory interactions with intracellular structural networks, and [PSI] prion may be formed as a by-product of this process. PMID- 10471704 TI - Dictyostelium myosin II G680V suppressors exhibit overlapping spectra of biochemical phenotypes including facilitated phosphate release. AB - We have biochemically characterized 13 intragenic suppressors of the G680V mutation of Dictyostelium myosin II. In the absence of the G680V mutation, the suppressors result in a number of deviant behaviors, most commonly an increase in the basal (actin-independent) ATPase of the motor. This phenotype is complementary to that of the G680V mutant and supports our proposal that the latter impairs phosphate release. Different subsets of the mutants also suffer from poor ATPase enhancement by 1 mg/ml actin, failure to release from actin in the presence of ATPgammaS (or ADP and salt), and excessive release from actin in the presence of ADP. The patterns of suppressor behaviors suggest that, in general, they are facilitating P(i)-releasing state(s) of the motor, but that different individual suppressors may secondarily perturb other states or actions of the motor. PMID- 10471705 TI - Functional overlap between the mec-8 gene and five sym genes in Caenorhabditis elegans. AB - Earlier work showed that the Caenorhabditis elegans gene mec-8 encodes a regulator of alternative RNA splicing and that mec-8 null mutants have defects in sensory neurons and body muscle attachment but are generally viable and fertile. We have used a genetic screen to identify five mutations in four genes, sym-1-sym 4, that are synthetically lethal with mec-8 loss-of-function mutations. The phenotypes of sym single mutants are essentially wild type. mec-8; sym-1 embryos arrest during embryonic elongation and exhibit defects in the attachment of body muscle to extracellular cuticle. sym-1 can encode a protein containing a signal sequence and 15 contiguous leucine-rich repeats. A fusion of sym-1 and the gene for green fluorescent protein rescued the synthetic lethality of mec-8; sym-1 mutants; the fusion protein was secreted from the apical hypodermal surface of the embryo. We propose that SYM-1 helps to attach body muscle to the extracellular cuticle and that another gene that is dependent upon mec-8 for pre mRNA processing overlaps functionally with sym-1. RNA-mediated interference experiments indicated that a close relative of sym-1 functionally overlaps both sym-1 and mec-8 in affecting muscle attachment. sym-2, sym-3, and sym-4 appear to provide additional functions that are essential in the absence of mec-8(+). PMID- 10471708 TI - Reduced sequence variability on the Neo-Y chromosome of Drosophila americana americana. AB - Sex chromosomes are generally morphologically and functionally distinct, but the evolutionary forces that cause this differentiation are poorly understood. Drosophila americana americana was used in this study to examine one aspect of sex chromosome evolution, the degeneration of nonrecombining Y chromosomes. The primary X chromosome of D. a. americana is fused with a chromosomal element that was ancestrally an autosome, causing this homologous chromosomal pair to segregate with the sex chromosomes. Sequence variation at the Alcohol Dehydrogenase (Adh) gene was used to determine the pattern of nucleotide variation on the neo-sex chromosomes in natural populations. Sequences of Adh were obtained for neo-X and neo-Y chromosomes of D. a. americana, and for Adh of D. a. texana, in which it is autosomal. No significant sequence differentiation is present between the neo-X and neo-Y chromosomes of D. a. americana or the autosomes of D. a. texana. There is a significantly lower level of sequence diversity on the neo-Y chromosome relative to the neo-X in D. a. americana. This reduction in variability on the neo-Y does not appear to have resulted from a selective sweep. Coalescent simulations of the evolutionary transition of an autosome into a Y chromosome indicate there may be a low level of recombination between the neo-X and neo-Y alleles of Adh and that the effective population size of this chromosome may have been reduced below the expected value of 25% of the autosomal effective size, possibly because of the effects of background selection or sexual selection. PMID- 10471709 TI - The Drosophila pumilio gene encodes two functional protein isoforms that play multiple roles in germline development, gonadogenesis, oogenesis and embryogenesis. AB - The pumilio (pum) gene plays an essential role in embryonic patterning and germline stem cell (GSC) maintenance during oogenesis in Drosophila. Here we report on a phenotypic analysis using pum(ovarette) mutations, which reveals multiple functions of pum in primordial germ cell proliferation, larval ovary formation, GSC division, and subsequent oogenic processes, as well as in oviposition. Specifically, by inducing pum(-) GSC clones at the onset of oogenesis, we show that pum is directly involved in GSC division, a function that is distinct from its requirement in primordial germ cells. Furthermore, we show that pum encodes 156- and 130-kD proteins, both of which are functional isoforms. Among pum(ovarette) mutations, pum(1688) specifically eliminates the 156-kD isoform but not the 130-kD isoform, while pum(2003) and pum(4277) specifically affect the 130-kD isoform but not the 156-kD isoform. Normal doses of both isoforms are required for the zygotic function of pum, yet either isoform alone at a normal dose is sufficient for the maternal effect function of pum. A pum cDNA transgene that contains the known open reading frame encodes only the 156-kD isoform and rescues the phenotype of both pum(1688) and pum(2003) mutants. These observations suggest that the 156- and 130-kD isoforms can compensate for each other's function in a dosage-dependent manner. Finally, we present molecular evidence suggesting that the two PUM isoforms share some of their primary structures. PMID- 10471710 TI - Recombination rate predicts inversion size in Diptera. AB - Most species of the Drosophila genus and other Diptera are polymorphic for paracentric inversions. A common observation is that successful inversions are of intermediate size. We test here the hypothesis that the selected property is the recombination length of inversions, not their physical length. If so, physical length of successful inversions should be negatively correlated with recombination rate across species. This prediction was tested by a comprehensive statistical analysis of inversion size and recombination map length in 12 Diptera species for which appropriate data are available. We found that (1) there is a wide variation in recombination map length among species; (2) physical length of successful inversions varies greatly among species and is inversely correlated with the species recombination map length; and (3) neither the among-species variation in inversion length nor the correlation are observed in unsuccessful inversions. The clear differences between successful and unsuccessful inversions point to natural selection as the most likely explanation for our results. Presumably the selective advantage of an inversion increases with its length, but so does its detrimental effect on fertility due to double crossovers. Our analysis provides the strongest and most extensive evidence in favor of the notion that the adaptive value of inversions stems from their effect on recombination. PMID- 10471706 TI - The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes. AB - A fundamental goal of genetics and functional genomics is to identify and mutate every gene in model organisms such as Drosophila melanogaster. The Berkeley Drosophila Genome Project (BDGP) gene disruption project generates single P element insertion strains that each mutate unique genomic open reading frames. Such strains strongly facilitate further genetic and molecular studies of the disrupted loci, but it has remained unclear if P elements can be used to mutate all Drosophila genes. We now report that the primary collection has grown to contain 1045 strains that disrupt more than 25% of the estimated 3600 Drosophila genes that are essential for adult viability. Of these P insertions, 67% have been verified by genetic tests to cause the associated recessive mutant phenotypes, and the validity of most of the remaining lines is predicted on statistical grounds. Sequences flanking >920 insertions have been determined to exactly position them in the genome and to identify 376 potentially affected transcripts from collections of EST sequences. Strains in the BDGP collection are available from the Bloomington Stock Center and have already assisted the research community in characterizing >250 Drosophila genes. The likely identity of 131 additional genes in the collection is reported here. Our results show that Drosophila genes have a wide range of sensitivity to inactivation by P elements, and provide a rationale for greatly expanding the BDGP primary collection based entirely on insertion site sequencing. We predict that this approach can bring >85% of all Drosophila open reading frames under experimental control. PMID- 10471707 TI - An exploration of the sequence of a 2.9-Mb region of the genome of Drosophila melanogaster: the Adh region. AB - A contiguous sequence of nearly 3 Mb from the genome of Drosophila melanogaster has been sequenced from a series of overlapping P1 and BAC clones. This region covers 69 chromosome polytene bands on chromosome arm 2L, including the genetically well-characterized "Adh region." A computational analysis of the sequence predicts 218 protein-coding genes, 11 tRNAs, and 17 transposable element sequences. At least 38 of the protein-coding genes are arranged in clusters of from 2 to 6 closely related genes, suggesting extensive tandem duplication. The gene density is one protein-coding gene every 13 kb; the transposable element density is one element every 171 kb. Of 73 genes in this region identified by genetic analysis, 49 have been located on the sequence; P-element insertions have been mapped to 43 genes. Ninety-five (44%) of the known and predicted genes match a Drosophila EST, and 144 (66%) have clear similarities to proteins in other organisms. Genes known to have mutant phenotypes are more likely to be represented in cDNA libraries, and far more likely to have products similar to proteins of other organisms, than are genes with no known mutant phenotype. Over 650 chromosome aberration breakpoints map to this chromosome region, and their nonrandom distribution on the genetic map reflects variation in gene spacing on the DNA. This is the first large-scale analysis of the genome of D. melanogaster at the sequence level. In addition to the direct results obtained, this analysis has allowed us to develop and test methods that will be needed to interpret the complete sequence of the genome of this species. Before beginning a Hunt, it is wise to ask someone what you are looking for before you begin looking for it. Milne 1926 PMID- 10471711 TI - A direct screen identifies new flight muscle mutants on the Drosophila second chromosome. AB - An ethyl methanesulfonate mutagenesis of Drosophila melanogaster was undertaken, and >3000 mutagenized second chromosomes were generated. More than 800 homozygous viable lines were established, and adults were screened directly under polarized light for muscle defects. A total of 16 mutant strains in which the indirect flight muscles were reduced in volume or disorganized or were otherwise abnormal were identified. These fell into seven recessive and one semidominant complementation groups. Five of these eight complementation groups, including the semidominant mutation, have been mapped using chromosomal deficiencies and meiotic recombination. Two complementation groups mapped close to the Myosin heavy chain gene, but they are shown to be in different loci. Developmental analysis of three mutations showed that two of these are involved in the early stages of adult myogenesis while the other showed late defects. This is the first report of results from a systematic and direct screen for recessive flight muscle defects. This mutant screen identifies genes affecting the flight muscles, which are distinct from those identified when screening for flightlessness. PMID- 10471712 TI - A genetic screen for modifiers of E2F in Drosophila melanogaster. AB - The activity of the E2F transcription factor is regulated in part by pRB, the protein product of the retinoblastoma tumor suppressor gene. Studies of tumor cells show that the p16(ink4a)/cdk4/cyclin D/pRB pathway is mutated in most forms of cancer, suggesting that the deregulation of E2F, and hence the cell cycle, is a common event in tumorigenesis. Extragenic mutations that enhance or suppress E2F activity are likely to alter cell-cycle control and may play a role in tumorigenesis. We used an E2F overexpression phenotype in the Drosophila eye to screen for modifiers of E2F activity. Coexpression of dE2F and its heterodimeric partner dDP in the fly eye induces S phases and cell death. We isolated 33 enhancer mutations of this phenotype by EMS and X-ray mutagenesis and by screening a deficiency library collection. The majority of these mutations sorted into six complementation groups, five of which have been identified as alleles of brahma (brm), moira (mor) osa, pointed (pnt), and polycephalon (poc). osa, brm, and mor encode proteins with homology to SWI1, SWI2, and SWI3, respectively, suggesting that the activity of a SWI/SNF chromatin-remodeling complex has an important impact on E2F-dependent phenotypes. Mutations in poc also suppress phenotypes caused by p21(CIP1) expression, indicating an important role for polycephalon in cell-cycle control. PMID- 10471713 TI - Soma-to-germline interactions during Drosophila oogenesis are influenced by dose sensitive interactions between cut and the genes cappuccino, ovarian tumor and agnostic. AB - The cut gene of Drosophila melanogaster encodes a homeodomain protein that regulates a soma-to-germline signaling pathway required for proper morphology of germline cells during oogenesis. cut is required solely in somatic follicle cells, and when cut function is disrupted, membranes separating adjacent nurse cells break down and the structural integrity of the actin cytoskeleton is compromised. To understand the mechanism by which cut expression influences germline cell morphology, we determined whether binucleate cells form by defective cytokinesis or by fusion of adjacent cells. Egg chambers produced by cut, cappuccino, and chickadee mutants contained binucleate cells in which ring canal remnants stained with antibodies against Hu-li tai shao and Kelch, two proteins that are added to ring canals after cytokinesis is complete. In addition, defects in egg chamber morphology were observed only in middle to late stages of oogenesis, suggesting that germline cell cytokineses were normal in these mutants. cut exhibited dose-sensitive genetic interactions with cappuccino but not with chickadee or other genes that regulate cytoskeletal function, including armadillo, spaghetti squash, quail, spire, Src64B, and Tec29A. Genomic regions containing genes that cooperate with cut were identified by performing a second-site noncomplementing screen using a collection of chromosomal deficiencies. Sixteen regions that interact with cut during oogenesis and eight regions that interact during the development of other tissues were identified. Genetic interactions between cut and the ovarian tumor gene were identified as a result of the screen. In addition, the gene agnostic was found to be required during oogenesis, and genetic interactions between cut and agnostic were revealed. These results demonstrate that a signaling pathway regulating the morphology of germline cells is sensitive to genetic doses of cut and the genes cappuccino, ovarian tumor, and agnostic. Since these genes regulate cytoskeletal function and cAMP metabolism, the cut-mediated pathway functionally links these elements to preserve the cytoarchitecture of the germline cells. PMID- 10471714 TI - Population structure among African and derived populations of Drosophila simulans: evidence for ancient subdivision and recent admixture. AB - Previous studies based on allozyme variation have found little evidence for genetic differentiation in Drosophila simulans. On the basis of DNA sequence variation at two nuclear loci in four African populations of D. simulans, we show that there is significant structure to D. simulans populations within Africa. Variation at one of the loci, vermilion, appears to be neutral and supports an eastern African origin for European and American populations. Samples from the West Indies, Europe, and North America had a nucleotide diversity lower than that of African populations at vermilion and show nonequilibrium haplotype distributions at both vermilion and G6pd, consistent with a hypothesis of recent bottleneck and possibly also admixture in the history of these populations. Directional selection, previously documented at G6pd, appears to have occurred within the coalescence time of the species, obscuring deep population history. PMID- 10471715 TI - Roles of the C terminus of Armadillo in Wingless signaling in Drosophila. AB - Drosophila melanogaster Armadillo and its vertebrate homolog beta-catenin play multiple roles during development. Both are components of cell-cell adherens junctions and both transduce Wingless (Wg)/Wnt intercellular signals. The current model for Wingless signaling proposes that Armadillo binds the DNA-binding protein dTCF, forming a bipartite transcription factor that activates Wingless responsive genes. In this model, Armadillo's C-terminal domain is proposed to serve an essential role as a transcriptional activation domain. In Xenopus, however, overexpression of C-terminally truncated beta-catenin activates Wnt signaling, suggesting that the C-terminal domain might not be essential. We reexamined the function of Armadillo's C terminus in Wingless signaling. We found that C-terminally truncated mutant Armadillo has a deficit in Wg-signaling activity, even when corrected for reduced protein levels. However, we also found that Armadillo proteins lacking all or part of the C terminus retain some signaling ability if overexpressed, and that mutants lacking different portions of the C-terminal domain differ in their level of signaling ability. Finally, we found that the C terminus plays a role in Armadillo protein stability in response to Wingless signal and that the C-terminal domain can physically interact with the Arm repeat region. These data suggest that the C-terminal domain plays a complex role in Wingless signaling and that Armadillo recruits the transcriptional machinery via multiple contact sites, which act in an additive fashion. PMID- 10471716 TI - RAPD-based genetic linkage maps of Tribolium castaneum. AB - A genetic map of the red flour beetle (Tribolium castaneum) integrating molecular with morphological markers was constructed using a backcross population of 147 siblings. The map defines 10 linkage groups (LGs), presumably corresponding to the 10 chromosomes, and consists of 122 randomly amplified polymorphic DNA (RAPD) markers, six molecular markers representing identified genes, and five morphological markers. The total map length is 570 cM, giving an average marker resolution of 4.3 cM. The average physical distance per genetic distance was estimated at 350 kb/cM. A cluster of loci showing distorted segregation was detected on LG9. The process of converting RAPD markers to sequence-tagged site markers was initiated: 18 RAPD markers were cloned and sequenced, and single strand conformational polymorphisms were identified for 4 of the 18. The map positions of all 4 coincided with those of the parent RAPD markers. PMID- 10471717 TI - Switch in codon bias and increased rates of amino acid substitution in the Drosophila saltans species group. AB - We investigated the nucleotide composition of five genes, Xdh, Adh, Sod, Per, and 28SrRNA, in nine species of Drosophila (subgenus Sophophora) and one of Scaptodrosophila. The six species of the Drosophila saltans group markedly differ from the others in GC content and codon use bias. The GC content in the third codon position, and to a lesser extent in the first position and the introns, is higher in the D. melanogaster and D. obscura groups than in the D. saltans group (in Scaptodrosophila it is intermediate but closer to the melanogaster and obscura species). Differences are greater for Xdh than for Adh, Sod, Per, and 28SrRNA, which are functionally more constrained. We infer that rapid evolution of GC content in the saltans lineage is largely due to a shift in mutation pressure, which may have been associated with diminished natural selection due to smaller effective population numbers rather than reduced recombination rates. The rate of GC content evolution impacts the rate of protein evolution and may distort phylogenetic inferences. Previous observations suggesting that GC content evolution is very limited in Drosophila may have been distorted due to the restricted number of genes and species (mostly D. melanogaster) investigated. PMID- 10471718 TI - Bioinvasions of the medfly Ceratitis capitata: source estimation using DNA sequences at multiple intron loci. AB - The Mediterranean fruit fly, Ceratitis capitata, is a devastating agricultural pest that threatens to become established in vulnerable areas such as California and Florida. Considerable controversy surrounds the status of Californian medfly infestations: Do they represent repeated introductions or the persistence of a resident population? Attempts to resolve this question using traditional population genetic markers and statistical methods are problematic because the most likely source populations in Latin America were themselves only recently colonized and are genetically very similar. Here, significant population structure among several New World medfly populations is demonstrated through the analysis of DNA sequence variation at four intron loci. Surprisingly, in these newly founded populations, estimates of population structure increase when measures of subdivision take into account the relatedness of alleles as well as their frequency. A nonequilibrium, likelihood-based statistical test that utilizes multilocus genotypes suggests that the sole medfly captured in California during 1996 was introduced from Latin America and was less likely to be a remnant of an ancestral Californian population. Many bioinvasions are hierarchical in nature, consisting of several sequential or overlapping invasion events, the totality of which can be termed a metainvasion. Phylogenetic data from multilocus DNA sequences will be vital to understanding the evolutionary and ecological processes that underlie metainvasions and to resolving their constituent levels. PMID- 10471719 TI - The catecholamines up (Catsup) protein of Drosophila melanogaster functions as a negative regulator of tyrosine hydroxylase activity. AB - We report the genetic, phenotypic, and biochemical analyses of Catecholamines up (Catsup), a gene that encodes a negative regulator of tyrosine hydroxylase (TH) activity. Mutations within this locus are semidominant lethals of variable penetrance that result in three broad, overlapping effective lethal phases (ELPs), indicating that the Catsup gene product is essential throughout development. Mutants from each ELP exhibit either cuticle defects or catecholamine-related abnormalities, such as melanotic salivary glands or pseudotumors. Additionally, Catsup mutants have significantly elevated TH activity that may arise from a post-translational modification of the enzyme. The hyperactivation of TH in Catsup mutants results in abnormally high levels of catecholamines, which can account for the lethality, visible phenotypes, and female sterility observed in these mutants. We propose that Catsup is a component of a novel system that downregulates TH activity, making Catsup the fourth locus found within the Dopa decarboxylase (Ddc) gene cluster that functions in catecholamine metabolism. PMID- 10471720 TI - Genetic dissection of X-linked interspecific hybrid placental dysplasia in congenic mouse strains. AB - Interspecific hybridization in the genus Mus results in male sterility and X linked placental dysplasia. We have generated several congenic laboratory mouse lines (Mus musculus) in which different parts of the maternal X chromosome were derived from M. spretus. A strict positive correlation between placental weight and length of the M. spretus-derived part of the X chromosome was shown. Detailed analysis was carried out with one congenic strain that retained a M. spretus interval between 12.0 and 30.74 cM. This strain consistently produced hyperplastic placentas that exhibited an average weight increase of 180% over the weight of control placentas. In derived subcongenic strains, however, increased placental weight could no longer be observed. Morphometric analysis of these placentas revealed persistence of abnormal morphology. Fully developed placental hyperplasia could be reconstituted by recombination of proximal and central M. spretus intervals with an intervening M. musculus region. These results may suggest that placental dysplasia of interspecific mouse hybrids is caused by multiple loci clustered on the X chromosome that act synergistically. Alternatively, it is possible that changes in chromatin structure in interspecific hybrids that influence gene expression are dependent on the length of the alien chromosome. PMID- 10471721 TI - Genomic organization of the S locus: Identification and characterization of genes in SLG/SRK region of S(9) haplotype of Brassica campestris (syn. rapa). AB - In Brassica, two self-incompatibility genes, encoding SLG (S locus glycoprotein) and SRK (S-receptor kinase), are located at the S locus and expressed in the stigma. Recent molecular analysis has revealed that the S locus is highly polymorphic and contains several genes, i.e., SLG, SRK, the as-yet-unidentified pollen S gene(s), and other linked genes. In the present study, we searched for expressed sequences in a 76-kb SLG/SRK region of the S(9) haplotype of Brassica campestris (syn. rapa) and identified 10 genes in addition to the four previously identified (SLG(9), SRK(9), SAE1, and SLL2) in this haplotype. This gene density (1 gene/5.4 kb) suggests that the S locus is embedded in a gene-rich region of the genome. The average G + C content in this region is 32.6%. An En/Spm-type transposon-like element was found downstream of SLG(9). Among the genes we identified that had not previously been found to be linked to the S locus were genes encoding a small cysteine-rich protein, a J-domain protein, and an antisilencing protein (ASF1) homologue. The small cysteine-rich protein was similar to a pollen coat protein, named PCP-A1, which had previously been shown to bind SLG. PMID- 10471722 TI - Arabidopsis PAI gene arrangements, cytosine methylation and expression. AB - Previous analysis of the PAI tryptophan biosynthetic gene family in Arabidopsis thaliana revealed that the Wassilewskija (WS) ecotype has four PAI genes at three unlinked sites: a tail-to-tail inverted repeat at one locus (PAI1-PAI4) plus singlet genes at two other loci (PAI2 and PAI3). The four WS PAI genes are densely cytosine methylated over their regions of DNA identity. In contrast, the Columbia (Col) ecotype has three singlet PAI genes at the analogous loci (PAI1, PAI2, and PAI3) and no cytosine methylation. To understand the mechanism of PAI gene duplication at the polymorphic PAI1 locus, and to investigate the relationship between PAI gene arrangement and PAI gene methylation, we analyzed 39 additional ecotypes of Arabidopsis. Six ecotypes had PAI arrangements similar to WS, with an inverted repeat and dense PAI methylation. All other ecotypes had PAI arrangements similar to Col, with no PAI methylation. The novel PAI methylated ecotypes provide insights into the mechanisms underlying PAI gene duplication and methylation, as well as the relationship between methylation and gene expression. PMID- 10471723 TI - Meiotic drive of chromosomal knobs reshaped the maize genome. AB - Meiotic drive is the subversion of meiosis so that particular genes are preferentially transmitted to the progeny. Meiotic drive generally causes the preferential segregation of small regions of the genome; however, in maize we propose that meiotic drive is responsible for the evolution of large repetitive DNA arrays on all chromosomes. A maize meiotic drive locus found on an uncommon form of chromosome 10 [abnormal 10 (Ab10)] may be largely responsible for the evolution of heterochromatic chromosomal knobs, which can confer meiotic drive potential to every maize chromosome. Simulations were used to illustrate the dynamics of this meiotic drive model and suggest knobs might be deleterious in the absence of Ab10. Chromosomal knob data from maize's wild relatives (Zea mays ssp. parviglumis and mexicana) and phylogenetic comparisons demonstrated that the evolution of knob size, frequency, and chromosomal position agreed with the meiotic drive hypothesis. Knob chromosomal position was incompatible with the hypothesis that knob repetitive DNA is neutral or slightly deleterious to the genome. We also show that environmental factors and transposition may play a role in the evolution of knobs. Because knobs occur at multiple locations on all maize chromosomes, the combined effects of meiotic drive and genetic linkage may have reshaped genetic diversity throughout the maize genome in response to the presence of Ab10. Meiotic drive may be a major force of genome evolution, allowing revolutionary changes in genome structure and diversity over short evolutionary periods. PMID- 10471724 TI - Maize R2R3 Myb genes: Sequence analysis reveals amplification in the higher plants. AB - Transcription factors containing the Myb-homologous DNA-binding domain are widely found in eukaryotes. In plants, R2R3 Myb-domain proteins are involved in the control of form and metabolism. The Arabidopsis genome harbors >100 R2R3 Myb genes, but few have been found in monocots, animals, and fungi. Using RT-PCR from different maize organs, we cloned 480 fragments corresponding to a 42-44 residue long sequence spanning the region between the conserved DNA-recognition helices (Myb(BRH)) of R2R3 Myb domains. We determined that maize expresses >80 different R2R3 Myb genes, and evolutionary distances among maize Myb(BRH) sequences indicate that most of the amplification of the R2R3 Myb gene family occurred after the origin of land plants but prior to the separation of monocots and dicots. In addition, evidence is provided for the very recent duplication of particular classes of R2R3 Myb genes in the grasses. Together, these findings render a novel line of evidence for the amplification of the R2R3 Myb gene family in the early history of land plants and suggest that maize provides a possible model system to examine the hypothesis that the expansion of Myb genes is associated with the regulation of novel plant cellular functions. PMID- 10471725 TI - Analysis and mapping of gene families encoding beta-1,3-glucanases of soybean. AB - Oligonucleotide primers designed for conserved sequences from coding regions of beta-1,3-glucanase genes from different species were used to amplify related sequences from soybean [Glycine max (L.) Merr.]. Sequencing and cross hybridization of amplification products indicated that at least 12 classes of beta-1,3-glucanase genes exist in the soybean. Members of classes mapped to 34 loci on five different linkage groups using an F(2) population of 56 individuals. beta-1,3-Glucanase genes are clustered onto regions of five linkage groups. Data suggest that more closely related genes are clustered together on one linkage group or on duplicated regions of linkage groups. Northern blot analyses performed on total RNA from root, stem, leaf, pod, flower bud, and hypocotyl using DNA probes for the different classes of beta-1,3-glucanase genes revealed that the mRNA levels of all classes were low in young leaves. SGlu2, SGlu4, SGlu7, and SGlu12 mRNA were highly accumulated in young roots and hypocotyls. SGlu7 mRNA also accumulated in pods and flower buds. PMID- 10471727 TI - The effects of deleterious mutations on linked, neutral variation in small populations. AB - The effects of recessive, deleterious mutations on genetic variation at linked neutral loci can be heterozygosity-decreasing because of reduced effective population sizes or heterozygosity-increasing because of associative overdominance. Here we examine the balance between these effects by simulating individual diploid genotypes in small panmictic populations. The haploid genome consists of one linkage group with 1000 loci that can have deleterious mutations and a neutral marker. Combinations of the following parameters are studied: gametic mutation rate to harmful alleles (U), population size (N), recombination rate (r), selection coefficient (s), and dominance (h). Tight linkage (r C73 and. These results directly show that tRNA identity contributes to the degree of complementarity to the transition state for tRNA charging in the active site of an aminoacyl-tRNA synthetase:aminoacyl adenylate:tRNA complex. PMID- 10471731 TI - Differential subcellular localization of human MutY homolog (hMYH) and the functional activity of adenine:8-oxoguanine DNA glycosylase. AB - The post-replicative adenine:8-oxo-7,8-dihydroguanine (GO) mismatch is crucial for G:C to T:A transversion. This mismatch is corrected by Escherichia coli MutY which excises the adenine from A:GO. A candidate gene coding for the human counterpart of MutY has been cloned as hMYH. However, the function and enzyme activities of the gene product have not been identified. We previously demonstrated that an epitope-tagged hMYH protein behaves as a mitochondrial protein. In the present study, we have identified an alternative hMYH transcript, termed type 2, which differs in the exon 1 sequence of the known transcript (type 1). A nuclear localization for the type 2 protein was revealed by detection of epitope-tagged protein in COS-7 cells. Expression of both type 1 and type 2 transcripts was reduced in post-mitotic tissues. hMYH cDNA suppressed the mutator phenotype of E.coli mutY. In vitro expressed hMYH showed adenine DNA glycosylase activity toward the A:GO substrate. The protein can bind to A:GO, and to T:GO and G:GO without apparent catalysis. These results represent the first demonstration of the function of the hMYH gene product which is differentially transported into the nucleus or the mitochondria by alternative splicing PMID- 10471732 TI - Escherichia coli RNA polymerase translocation is accompanied by periodic bending of the DNA. AB - RNA polymerase was halted in consecutive registers of RNA synthesis ranging from registers 11 to 68. Non-denaturing gel electrophoresis shows that the mobility of the complexes varies (up to 15%), indicating that halted complexes differ in their conformation. The electrophoretic mobility changes with an approximate 10 register periodicity. The change of the mobility can be attributed to relative changes of RNA polymerase-induced bending angle. We suggest that the periodicity of the bending angle reflects periodic changes of the conformation of the halted complexes that might have relevance for the translocation mechanism. PMID- 10471733 TI - Role of the locus and of the resistance gene on gene amplification frequency in methotrexate resistant Leishmania tarentolae. AB - The protozoan parasite Leishmania resists the antifolate methotrexate (MTX) by amplifying the R locus dihydrofolate reductase-thymidylate synthase ( dhfr-ts ) gene, the H locus ptr1 pterin reductase gene, and finally by mutation in a common folate/MTX transporter. Amplification of dhfr-ts has never been observed in Leishmania tarentolae MTX resistant mutants while ptr1 amplification is common. We have selected a L.tarentolae ptr1 null mutant for MTX resistance and observed dhfr-ts amplification in this mutant demonstrating that once a preferred resistance mechanism is unavailable, a second one will take over. By introducing the ptr1 gene at the R locus and the dhfr-ts gene at the H locus by gene targeting, we investigated the role of the resistance gene and the locus on the rate of gene amplification. Transfection studies indicated that ptr1 gave higher levels of MTX resistance than dhfr-ts. Consistent with this, when ptr1 was present as part of either the H locus or the R locus it was invariably amplified, while dhfr-ts was only amplified when ptr1 was inactivated. When dhfr-ts was present in a ptr1 null background on both the H locus and the R locus, amplification from the H locus was more frequent suggesting that both the gene and the locus are determining the frequency of gene amplification in Leishmania. PMID- 10471734 TI - The 5'-untranslated region of GM-CSF mRNA suppresses translational repression mediated by the 3' adenosine-uridine-rich element and the poly(A) tail. AB - Granulocyte-macrophage colony stimulating factor (GM-CSF) mRNA levels are controlled post-transcriptionally by the 3'-untranslated region (UTR) adenosine uridine-rich element (ARE). In untransformed, resting cells, the ARE targets GM CSF mRNA for rapid degradation, thereby significantly suppressing protein expression. We used a rabbit reticulocyte lysate (RRL) cell-free system to examine translational regulation of GM-CSF expression. We uncoupled decay rates from rates of translation by programming the RRL with an excess of mRNAs. Capped, full-length, polyadenyl-ated human GM-CSF mRNA (full-length 5'-UTR AUUUA+A90) and an ARE-modified version (full-length 5'-UTR AUGUA+A90) produced identical amounts of protein. When the 5'-UTR was replaced with an irrelevant synthetic leader sequence (syn 5'-UTR), translation of syn 5'-UTR AUUUA+A90 mRNA was suppressed by >20-fold. Mutation of the ARE or removal of the poly(A) tail relieved this inhibition. Thus, in the absence of a native 5'-UTR, the ARE and poly(A) tail act in concert to block GM-CSF mRNA translation. Substitutions of different regions of the native 5'-UTR revealed that the entire sequence was essential in maintaining the highest rates of translation. However, shorter 10-12 nt contiguous 5'-UTR regions supported 50-60% of maximum translation. The 5'-UTR is highly conserved, suggesting similar regulation in multiple species and in these studies was the dominant element regulating GM-CSF mRNA translation, overriding the inhibitory effects of the ARE and the poly(A) tail. PMID- 10471735 TI - An NMR and mutational analysis of an RNA pseudoknot of Escherichia coli tmRNA involved in trans-translation. AB - Transfer-messenger RNA (tmRNA) is a unique molecule that combines properties from both tRNA and mRNA, and facilitates a novel translation reaction termed trans translation. According to phylogenetic sequence analysis among various bacteria and chemical probing analysis, the secondary structure of the 350-400 nt RNA is commonly characterized by a tRNA-like structure, and four pseudoknots with different sizes. A mutational analysis using a number of Escherichia coli tmRNA variants as well as a chemical probing analysis has recently demonstrated not only the presence of the smallest pseudoknot, PK1, upstream of the internal coding region, but also its direct implication in trans -translation. Here, NMR methods were used to investigate the structure of the 31 nt pseudoknot PK1 and its 11 mutants in which nucleotide substitutions are introduced into each of two stems or the linking loops. NMR results provide evidence that the PK1 RNA is folded into a pseudoknot structure in the presence of Mg(2+). Imino proton resonances were observed consistent with formation of two helical stem regions and these stems stacked to each other as often seen in pseudoknot structures, in spite of the existence of three intervening nucleo-tides, loop 3, between the stems. Structural instability of the pseudoknot structure, even in the presence of Mg(2+), was found in the PK1 mutants except in the loop 3 mutants which still maintained the pseudoknot folding. These results together with their biological activities indicate that trans -translation requires the pseudoknot structure stabilized by Mg(2+)and specific residues G61 and G62 in loop 3. PMID- 10471736 TI - Crosslinking of proteins to DNA in human nuclei using a 60 femtosecond 266 nm laser. AB - We developed appropriate conditions to use a laser with 60 femtosecond pulses, a frequency of 1 KHz and a wavelength of 266 nm to efficiently crosslink proteins to DNA in human nuclei for the purpose of using immunoprecipitation to study the binding of specific proteins to specific sequences of DNA under native conditions. Irradiation of nuclei for 30 min with 1-3 GW/cm(2)pulses crosslinked 10-12% of total protein to DNA. The efficiency of crosslinking was dose and protein specific. Histones H1 and H3 were crosslinked by 15 min of irradiation with 20-25% efficiency, at least 10 times more strongly than the other histones, consistent with experiments using conventional UV light. Irradiation for 15 min did not damage proteins, as assayed by SDS-PAGE of Ku-70 and histones. Although the same level of irradiation did not cause double-strand breaks, it did make the DNA partially insensitive to Eco RI restriction enzyme, probably through formation of thymidine dimers. Immuno-analysis of crosslinked nucleoprotein showed that Ku crosslinking to nuclear DNA is detectable only in the presence of breaks in the DNA, and that nucleosomes are bound to a significant fraction of the telomeric repeat (TTAGGG) (n). PMID- 10471737 TI - Messenger RNAs encoding mouse histone macroH2A1 isoforms are expressed at similar levels in male and female cells and result from alternative splicing. AB - Two protein isoforms of histone macroH2A1 (mH2A1) are found in mammalian cells. One isoform, mH2A1.2 is highly concentrated on the heterochromatinized inactive X chromosome (Xi) of female cells. mH2A1.2 protein is also present in male cells, but fails to form dense concentrations. Another protein isoform, mH2A1.1, differs from mH2A1.2 by a single short segment of amino acids. In this study, we cloned and characterized the genomic locus of the mouse mH2A1 gene and mapped it to chromosome 13. Two alternatively spliced transcripts derived from the mH2A1 locus are responsible for the generation of the two mH2A1 protein isoforms with mH2A1.2 mRNA being the most abundant spliced form in all tissues examined. The absolute amount of mH2A1 mRNA is similar in male and female cells for most tissues with the exception of testes where it is par-ticularly abundant. Both spliced forms are present in all adult tissues analyzed as well as in female embryonic stem cells. In contrast, male embryonic stem cells expressed mH2A1.1 at low levels if at all. The relatively abundant expression of mH2A1 in both sexes suggests that mH2A1 has functions in addition to a possible involvement in X chromosome inactivation. PMID- 10471738 TI - Repression of IS200 transposase synthesis by RNA secondary structures. AB - The IS 200 transposase, a 16 kDa polypeptide encoded by the single open reading frame (ORF) of the insertion element, has been identified using an expression system based on T7 RNA polymerase. In wild-type IS 200, two sets of internal inverted repeats that generate RNA secondary structures provide two independent mechanisms for repression of transposase synthesis. The inverted repeat located near the left end of IS 200 is a transcriptional terminator that terminates read through transcripts before they reach the IS 200 ORF. The terminator is functional in both directions and may terminate >80% of transcripts. Another control operates at the translational level: transposase synthesis is inhibited by occlusion of the ribosome-binding site (RBS) of the IS 200 ORF. The RBS (5' AGGGG-3') is occluded by formation of a mRNA stem-loop structure whose 3' end is located only 3 nt upstream of the start codon. This mechanism reduces transposase synthesis approximately 10-fold. Primer extension experiments with AMV reverse transcriptase have provided evidence that this stem-loop RNA structure is actually formed. Tight repression of transposase synthesis, achieved through synergistic mechanisms of negative control, may explain the unusually low transposition frequency of IS 200. PMID- 10471740 TI - Tolerance of 5-fluorodeoxyuridine resistant human thymidylate synthases to alterations in active site residues. AB - Fluoropyrimidines, such as 5-fluorouracil (5-FU), are used extensively in cancer therapy. In the cell, 5-FU is metabolized to 5-fluorodeoxyuridylate (5-FdUMP), a tight binding covalent inhibitor of thymidylate synthase (TS). In order to create 5-FdUMP resistant enzymes to protect chemosensitive normal cells and further understand mechanisms of 5-FdUMP resistance, we have randomized four residues within the active site of TS. Our previous studies identified alterations in residues which produce active TS with enhanced resistance to 5-fluorouridine (5 FdUR). By remutagenizing a subset of the 13 previously targeted residues (A197, L198, C199 and V204), an unbiased random library can be created allowing for extensive testing of all possible amino acid substitutions at each of the sites. Using genetic complementation and selection in Escherichia coli, we identified the spectrum of substitutions that yield active TS as well as those that resulted in 5-FdUR resistant mutants of TS. The 5-FdUR resistant TS were found to share several structural features including hydrophobic substitutions at residue 197, retention of the wild-type leucine 198, the alteration C199L (present in 64% of the drug-resistant library), and polar alterations of valine 204. The catalytic activity of mutants with these features was approximately equal to that of the wild-type TS. PMID- 10471739 TI - Hypersensitive substrate for ribonucleases. AB - A substrate for a hypersensitive assay of ribonucleolytic activity was developed in a systematic manner. This substrate is based on the fluorescence quenching of fluorescein held in proximity to rhodamine by a single ribonucleotide embedded within a series of deoxynucleotides. When the substrate is cleaved, the fluorescence of fluorescein is manifested. The optimal substrate is a tetranucleotide with a 5',6-carboxyfluorescein label (6-FAM) and a 3',6-carboxy tetramethylrhodamine (6-TAMRA) label: 6-FAM-dArUdAdA-6-TAMRA. The fluorescence of this substrate increases 180-fold upon cleavage. Bovine pancreatic ribonuclease A (RNase A) cleaves this substrate with a k (cat)/ K (m)of 3.6 x 10(7)M(-1)s(-1). Human angiogenin, which is a homolog of RNase A that promotes neovascularization, cleaves this substrate with a k (cat)/ K (m)of 3. 3 x 10(2)M(-1)s(-1). This value is >10-fold larger than that for other known substrates of angio-genin. With these attributes, 6-FAM-dArUdAdA-6-TAMRA is the most sensitive known substrate for detecting ribo-nucleolytic activity. This high sensitivity enables a simple protocol for the rapid determination of the inhibition constant ( K (i)) for competitive inhibitors such as uridine 3'-phosphate and adenosine 5'-diphos phate. PMID- 10471741 TI - Single-nucleotide patch base excision repair of uracil in DNA by mitochondrial protein extracts. AB - Mammalian mitochondria contain several 16.5 kb circular DNAs (mtDNA) encoding electron transport chain proteins. Reactive oxygen species formed as byproducts from oxidative phosphorylation in these organelles can cause oxidative deamination of cytosine and lead to uracil in mtDNA. Upon mtDNA replication, these lesions, if unrepaired, can lead to mutations. Until recently, it was thought that there was no DNA repair in mitochondria, but lately there is evidence that some lesions are efficiently repaired in these organelles. In the study of nuclear DNA repair, the in vitro repair measurements in cell extracts have provided major insights into the mechanisms. The use of whole-cell extract based DNA repair methods has revealed that mammalian nuclear base excision repair (BER) diverges into two pathways: the single-nucleotide replacement and long patch repair mechanisms. Similar in vitro methods have not been available for the study of mitochondrial BER. We have established an in vitro DNA repair system supported by rat liver mitochondrial protein extract and DNA substrates containing a single uracil opposite to a guanine. Using this approach, we examined the repair pathways and the identity of the DNA polymerase involved in mitochondrial BER (mtBER). Employing restriction analysis of in vitro repaired DNA to map the repair patch size, we demonstrate that only one nucleotide is incorporated during the repair process. Thus, in contrast to BER in the nucleus, mtBER of uracil in DNA is solely accomplished by single-nucleotide replacement. PMID- 10471742 TI - Identification of a mammalian RNA polymerase I holoenzyme containing components of the DNA repair/replication system. AB - Traditional models for transcription initiation by RNA polymerase I include a stepwise assembly of basic transcription factors/regulatory proteins on the core promoter to form a preinitiation complex. In contrast, we have identified a preassembled RNA polymerase I (RPI) complex that contains all the factors necessary and sufficient to initiate transcription from the rDNA promoter in vitro. The purified RPI holoenzyme contains the RPI homolog of TFIID, SL-1 and the rDNA transcription terminator factor (TTF-1), but lacks UBF, an activator of rDNA transcription. Certain components of the DNA repair/replication system, including Ku70/80, DNA topoisomerase I and PCNA, are also associated with the RPI complex. We have found that the holo-enzyme supported specific transcription and that specific transcription was stimulated by the RPI transcription activator UBF. These results support the hypothesis that a fraction of the RPI exists as a preassembled, transcriptionally competent complex that is readily recruited to the rDNA promoter, i.e. as a holoenzyme, and provide important new insights into the mechanisms governing initiation by RPI. PMID- 10471743 TI - An Arabidopsis cDNA encoding a DNA-binding protein that is highly similar to the DEAH family of RNA/DNA helicase genes. AB - A cDNA encoding a putative RNA and/or DNA helicase has been isolated from Arabidopsis thaliana cDNA libraries. The cloned cDNA is 5166 bases long, and its largest open reading frame encodes 1538 amino acids. The central region of the predicted protein is homologous to a group of nucleic acid helicases from the DEAD/H family. However, the N- and C-terminal regions of the Arabidopsis cDNA product are distinct from these animal DEIH proteins. We have found that the C terminal region contains three characteristic sequences: (i) two DNA-binding segments that form a probe helix (PH) involved in DNA recognition; (ii) an SV40 type nuclear localization signal; and (iii) 11 novel tandem-repeat sequences each consisting of about 28 amino acids. We have designated this cDNA as NIH (nuclear DEIH-boxhelicase). Functional character-ization of a recombinant fusion product containing the repeated region indicates that NIH may form homodimers, and that this is the active form in solution. Based on this information and the observation that the sequence homology is limited to the DEAH regions, we conclude that the biological roles of the plant helicase NIH differ from those of the animal DEIH family. PMID- 10471744 TI - Analysis of a YAC with human telomeres and oriP from epstein-barr virus in yeast and 293 cells. AB - One approach to the construction and propagation of a mammalian artificial chromosome is to build it up in Saccharomyces cerevisiae, using a yeast artificial chromosome (YAC) base. We have demonstrated that circular YACs carrying the Epstein-Barr virus origin of plasmid replication ( oriP ) are maintained as stable, episomal elements in human cells. We wished to determine whether this technology could be extended, to generate linear extrachromosomal elements. Here, we describe the generation of retrofitting constructs, which permit the addition of human telomeres and the oriP domain to YACs. The constructs contain 0.8 kb of human telomere sequence separated by a unique Not I site from 0.7 kb of Tetrahymena telomere sequence. These constructs seed telomere formation with approximately 40-60% efficiency in human 293-EBNA and HT1080 cells whether or not the Tetrahymena sequence is removed by Not I digestion. A detailed analysis demonstrates that YACs carrying the human telomere cassettes on both arms show instability of the telomere sequences in S.cerevisiae at a frequency of approximately 50%. Introduction of correctly retrofitted, linear oriP YACs into human 293-EBNA cells by lipofection resulted in the generation of circular extrachromosomal elements varying in size from 8 to 300 kb. However, no apparently linear YACs could be detected, suggesting that extrachromosomal maintenance of DNA with the oriP /EBNA-1 system is not compatible with linear molecules capped by telomeres. PMID- 10471745 TI - Sex-specific organisation of middle repetitive DNA sequences in the mealybug Planococcus lilacinus. AB - Differential organisation of homologous chromosomes is related to both sex determination and genomic imprinting in coccid insects, the mealybugs. We report here the identification of two middle repetitive sequences that are differentially organised between the two sexes and also within the same diploid nucleus. These two sequences form a part of the male-specific nuclease-resistant chromatin (NRC) fraction of a mealybug Planococcus lilacinus. To understand the phenomenon of differential organisation we have analysed the components of NRC by cloning the DNA sequences present, deciphering their primary sequence, nucleosomal organisation, genomic distri-bution and cytological localisation. Our observations suggest that the middle repetitive sequences within NRC are functionally significant and we discuss their probable involvement in male specific chromatin organisation. PMID- 10471746 TI - A conserved motif N-terminal to the DNA-binding domains of myogenic bHLH transcription factors mediates cooperative DNA binding with pbx-Meis1/Prep1. AB - The t(1;19) chromosomal translocation of pediatric pre-B cell leukemia produces chimeric oncoprotein E2a-Pbx1, which contains the N-terminal transactivation domain of the basic helix-loop-helix (bHLH) transcription factor, E2a, joined to the majority of the homeodomain protein, Pbx1. There are three Pbx family members, which bind DNA as heterodimers with both broadly expressed Meis/Prep1 homeo-domain proteins and specifically expressed Hox homeodomain proteins. These Pbx heterodimers can augment the function of transcriptional activators bound to adjacent elements. In heterodimers, a conserved tryptophan motif in Hox proteins binds a pocket on the surface of the Pbx homeodomain, while Meis/Prep1 proteins bind an N-terminal Pbx domain, raising the possibility that the tryptophan interaction pocket of the Pbx component of a Pbx-Meis/Prep1 complex is still available to bind trypto-phan motifs of other transcription factors bound to flanking elements. Here, we report that Pbx-Meis1/Prep1 binds DNA cooperatively with heterodimers of E2a and MyoD, myogenin, Mrf-4 or Myf-5. As with Hox proteins, a highly conserved tryptophan motif N-terminal to the DNA-binding domains of each myogenic bHLH family protein is required for cooperative DNA binding with Pbx-Meis1/Prep1. In vivo, MyoD requires this tryptophan motif to evoke chromatin remodeling in the Myogenin promoter and to activate Myogenin transcription. Pbx-Meis/Prep1 complexes, therefore, have the potential to cooperate with the myogenic bHLH proteins in regulating gene transcription. PMID- 10471747 TI - Drosophila and human RecQ5 exist in different isoforms generated by alternative splicing. AB - Members of the RecQ helicase superfamily have been implicated in DNA repair, recombination and replication. Although the genome of the budding yeast Saccharomyces cerevisiae encodes only a single member of this family, there are at least five human RecQ-related genes: RecQL, BLM, WRN, RecQ4 and RecQ5. Mutations in at least three of these are associated with diseases involving a predisposition to malignancies and a cellular phenotype that includes increased chromosome instability. Metazoan RecQ helicases are defined by a core region with characteristic helicase motifs and sequence similarity to Escherichia coli RecQ protein. This core region is typically flanked by extensive, highly charged regions, of largely unknown function. The recently reported human RecQ5, however, has only the core RecQ-homologous region. We describe here the identification of the Drosophila RecQ5 gene. We recovered cDNAs corresponding to three alternative splice forms of the RecQ5 transcript. Two of these generate nearly identical 54 kDa proteins that, like human RecQ5, consist of the helicase core only. The third splice variant encodes a 121 kDa isoform that, like other family members, has a C terminal extension rich in charged residues. A combination of RACE and cDNA analysis of human RECQ5 demonstrates extensive alternative splicing for this gene also, including some forms lacking helicase motifs and other conserved regions. PMID- 10471748 TI - Recombination and chimeragenesis by in vitro heteroduplex formation and in vivo repair. AB - We describe a simple method for creating libraries of chimeric DNA sequences derived from homologous parental sequences. A heteroduplex formed in vitro is used to transform bacterial cells where repair of regions of non-identity in the heteroduplex creates a library of new, recombined sequences composed of elements from each parent. Heteroduplex recombination provides a convenient addition to existing DNA recombination methods ('DNA shuffling') and should be particularly useful for recombining large genes or entire operons. This method can be used to create libraries of chimeric polynucleotides and proteins for directed evolution to improve their properties or to study structure-function relationships. We also describe a simple test system for evaluating the performance of DNA recombination methods in which recombination of genes encoding truncated green fluorescent protein (GFP) reconstructs the full-length gene and restores its characteristic fluorescence. Comprising seven truncated GFP constructs, this system can be used to evaluate the efficiency of recombination between mismatches separated by as few as 24 bp and as many as 463 bp. The optimized heteroduplex recombination protocol is quite efficient, generating nearly 30% fluorescent colonies for recombination between two genes containing stop codons 463 bp apart (compared to a theoretical limit of 50%). PMID- 10471749 TI - Polymorphism analysis and gene detection by minisequencing on an array of gel immobilized primers. AB - Two procedures, multibase and multiprimer, have been developed for single nucleotide extension of primers immobilized within polyacrylamide gel pads on a microchip. In the multibase assay, a primer is next to a polymorphic nucleotide; the nucleotide is identified by the specificity with which the primer incorporates fluorescently labeled dideoxyribo-nucleoside triphosphates. In the multiprimer assay, several primers containing different 3'-terminal nucleotides overlapping the variable nucleotide in DNA are used. The polymorphic nucleotide is identified according to the primer that is extended. The methods were compared for diagnosis of beta-thalassemia mutations. Isothermal amplification of the fluorescent signal was achieved by performing both assays at elevated temperature. Anthrax toxin genes were identified in a model system using this amplification method. PMID- 10471750 TI - Selective detection of ribose-methylated nucleotides in RNA by a mass spectrometry-based method. AB - Post-transcriptional methylation of ribose at position O-2' is one of the most common and conserved types of RNA modification. Details of the functional roles of these methylations are far from clear, although in tRNA they are involved at position 34 in regulation of codon recognition and in eukaryotic rRNAs they are required for subunit assembly. Experimental difficulties in the mapping of ribose methylations increase with RNA molecular size and the complexity of mixtures resulting from nuclease digestion. A new and relatively rapid approach based on tandem mass spectrometry is described in which any of four ion reaction pathways occurring in the mass spectrometer can be monitored which are highly specific for the presence of 2'-O -methylribose residues. These pathways emanate from further dissociation of ribose-methylated mononucleotide (Nmp) ions formed in the electrospray ionization region of the mass spectrometer to then form the base, methylribose phosphate or PO(3)(-)anions. The mass spectrometer can be set for detection of generic ribose methylation (Nm) in oligonucleotides, selectively for each of the common methylated nucleo-sides Cm, Gm, Am or Um or for specific cases in which the base or sugar is further modified. By direct combination of mass spectrometry with liquid chromatography the method can be applied to analysis of complex mixtures of oligonucleotides, as for instance from synthetic or in vitro reaction mixtures or from nuclease digests of RNA. An example is given in which the single ribose-methylated nucleoside in Escherichia coli 16S rRNA (1542 nt), N(4),O-2'-dimethylcytidine, is detected in 25 pmol of a RNase T1 digest and localized to the fragment 1402-CCCGp-1405 in a single 45 min analysis. PMID- 10471751 TI - Segmental genomic replacement in embryonic stem cells by double lox targeting. AB - We have applied Cre-mediated double lox recombination to embryonic stem (ES) cells to facilitate repeated knock-ins at a target locus, thus helping to assure correct temporal and spatial transgene expression in mice. Using homologous recombination, we inserted a double lox cassette a few nucleo-tides before the authentic ATG start of MHL-1, the gene coding for the major subunit of the asialoglycoprotein receptor. The cassette carries a marker gene bounded by heterospecific lox sites that cannot recombine with each other, but which can undergo recombination with like sites on an incoming double lox targeting vector. Cre-mediated replacement of the lox-delimited genomic segment at MHL-1 with targeting DNA occurs at a frequency three times that of random DNA integration and is sufficiently robust that correctly targeted ES colonies can be identified by PCR screening without relying on any drug selection procedure. Moreover, double lox recombination at the MHL-1 locus is not dependent on the presence or absence of a transcriptionally active promoter at the genomic target. The strategy and vectors described here are generally applicable to designing double lox targeted knock-ins at any locus in ES cells and should prove useful in more precise molecular engineering of the mouse genome. PMID- 10471752 TI - Tandem arrayed ligation of expressed sequence tags (TALEST): a new method for generating global gene expression profiles. AB - We have developed a new and simple method for quantitatively analyzing global gene expression profiles from cells or tissues. The process, called TALEST, or tandem arrayed ligation of expressed sequence tags, employs an oligonucleotide adapter containing a type IIs restriction enzyme site to facilitate the generation of short (16 bp) ESTs of fixed position in the mRNA. These ESTs are flanked by GC-clamped punctuation sequences which render them resistant to thermal denaturation, allowing their concatenation into long arrays and subsequent recognition and analysis by high-throughput DNA sequencing. A major advantage of the TALEST technique is the avoidance of PCR in all stages of the process and hence the attendant sequence-specific amplification biases that are inherent in other gene expression profiling methods such as SAGE, Differential Display, AFLP, etc. which rely on PCR. PMID- 10471753 TI - Regulation of average length of complex PCR product. AB - A method to achieve the preference towards longer products during PCR is described. The extent of this preference can be adjusted by slight variation of the PCR conditions. Being combined with the natural tendency of PCR to amplify shorter fragments more efficiently than longer ones, it allows one to regulate the average length of the complex PCR product over a very wide range to make it most suitable for further manipulations. The technique can be used for amplifying any complex DNA sample. PMID- 10471754 TI - Juxtaglomerular cell tumor with retroperitoneal fibrosis and secondary immune complex glomerulonephritis: a possible contribution of the renin angiotensin system to renal fibrosis. AB - We present a case of a 25-year-old woman with a renin-secreting juxtaglomerular cell tumor, retroperitoneal fibrosis associated with glomerular hypertrophy, glomerulonephritis, and marked tubulointerstitial alterations. Myofibroblasts, as shown by positive immunostaining for alpha-smooth muscle actin, were found along with transforming growth factor-beta (TGF-beta) in the interstitium of the tumor free kidney. Regarding the pathogenesis of renal fibrosis and glomerular hypertrophy, this case may provide evidence not only experimentally but also clinically that the renin-angiotensin system plays an important role because angiotensin II is known to induce renal fibrosis associated with increased TGF beta and the appearance of myofibroblasts. PMID- 10471755 TI - Development of focal segmental glomerulosclerosis in the renal allograft of a patient with lupus. AB - Nephritis has been a recognized complication of systemic lupus erythematosus since the early 1900s. Almost all lupus patients have some degree of renal involvement related to their condition, but a considerably smaller proportion of these patients actually progress to end-stage renal disease (ESRD). However, lupus patients are also susceptible to other primary renal insults that may significantly contribute to the deterioration in their renal function. We present a case of a patient with clinical and pathological evidence of lupus nephritis that progressed to ESRD and subsequently developed "recurrent" focal segmental glomerulosclerosis in her transplant kidney. Retrospective clinicopathologic correlation suggested the possibility of more than 1 primary renal process that eventually led to her dialysis-dependent state. This case illustrates the importance of meticulously examining both clinical and renal biopsy data in patients with lupus nephritis and considering the presence of co-existing renal pathologies to resolve an otherwise discordant picture of disease progression. These considerations may have important therapeutic and prognostic implications. PMID- 10471756 TI - Treatment of acute c-ANCA-positive vasculitis with mycophenolate mofetil. AB - Acute cytoplasmic antineutrophil cytoplasmic antibody (c-ANCA)-positive vasculitis is usually treated with cyclophosphamide and corticosteroids. The incidence of cyclophosphamide-induced lung injury, a potentially life-threatening event, is about 1%. We report on a patient with a history of cyclophosphamide induced lung injury 2 months after initial treatment of systemic c-ANCA-positive vasculitis. Six months later, the patient presented with acute renal failure caused by an acute relapse of vasculitis. Mycophenolate mofetil (MMF) is a potent immunosuppressive drug that recently has been shown to be effective in the maintenance therapy of c-ANCA-positive systemic vasculitis. With the patient's informed consent, we started treatment with MMF in combination with corticosteroids. Subsequently, anti-proteinase-3-titer (anti-Pr3-titer) returned to normal and renal function improved. In conclusion, MMF in combination with corticosteroids may be useful in the treatment of acute c-ANCA-positive vasculitis. PMID- 10471757 TI - Reversible bilateral hydronephrosis without obstruction in hepatitis B-associated polyarteritis nodosa. AB - The manifestations of polyarteritis nodosa (PAN) are varied, but urological abnormalities other than ureteric stenosis and orchitis have not been described. We report a case of hepatitis B-associated PAN with bilateral hydronephrosis without obstruction. Retrograde urography conclusively demonstrated the absence of obstruction. Vasculitis-related myopathy, or neuropathy of the ureter, is the most likely cause of this finding. The patient was treated with high-dose steroids, cyclophosphamide, and plasmapheresis with resolution of hydronephrosis. Although the patient required dialysis at initiation of therapy, she went on to recover sufficient renal function to discontinue dialysis. We review the literature on the treatment of hepatitis B-associated PAN and discuss the pitfalls in diagnosis of this condition. PMID- 10471758 TI - Acute renal failure in a patient with Rosai-Dorfman disease. AB - Acute renal failure developed in a 57-year-old woman who had Rosai-Dorfman disease diagnosed 1 year previously on a cervical lymph node. Organ imaging showed diffuse masses infiltrating both kidneys. The renal biopsy showed a lymphoplasmacytic and histiocytic process extensively replacing the parenchyma, which is in keeping with Rosai-Dorfman disease of the kidneys. However, the typical lymphophagocytic cells were lacking. This case illustrates that diagnosis of Rosai-Dorfman disease in renal biopsy can be very difficult, requiring both exclusion of many benign and malignant lesions and a high index of suspicion for this condition. In particular, lymphoma was excluded based on the mixed polyclonal composition of inflammatory cells and the absence of atypical lymphoid proliferation. The renal function partially recovered after a course of therapy combining VP-16 (etoposide) and dexamethasone and remained stable over 4-year follow-up. This report emphasizes the importance of early diagnosis and intervention to safeguard renal function in extensive Rosai-Dorfman disease. PMID- 10471759 TI - Biology of bone grafts. AB - The terminology of bone transplantation is complicated by the various histologic types of bone grafts. A graft moved from one site to another within the same individual is an autograft; the corresponding adjective is autologous or autogenous. An allograft (adjective, allogeneic) is tissue transferred between two genetically different individuals of the same species. A xenograft (adjective, xenogeneic) is tissue from one species implanted into a member of a different species. PMID- 10471760 TI - Biomechanical issues in bone transplantation. AB - Because the biomechanical competence of the graft is a central issue, many research studies of bone grafts include a biomechanical component. The biomechanical evaluation serves as a bottom-line measure of the experimental outcome in which some measure of mechanical performance is compared among treatment groups. This article considers the biomechanics of grafts in the context of this experimental work and focuses on three issues: (1) the interplay between biology and biomechanics of grafts, (2) the effects of treatment choices on biomechanical properties, and (3) model factors that may influence biomechanical performance. PMID- 10471761 TI - Bone banking. Update on methods and materials. AB - Bone allografts are being used in increasing numbers by orthopedic surgeons, yet many surgeons are unfamiliar with their preparation and processing, as well as their use as safe and effective transplants. This article reviews current sources of bone allografts, new methods of processing to achieve optimal results, the biology of incorporation of allografts, and new bone substitutes. PMID- 10471762 TI - Allograft bone. The influence of processing on safety and performance. AB - Advances in tissue processing technology have been important for the successful use of bone allografts. The challenge is to prepare allografts that are well cleaned, sterile, and free of viruses while still preserving the natural biologic and biomechanical properties of the tissue. This article discusses how processing techniques aimed at achieving safety and sterility can affect the properties vital for graft incorporation and healing. PMID- 10471763 TI - Long-term follow-up of patients with osteochondral allografts. A correlation between immunologic responses and clinical outcome. AB - This article confirms immunologic responses in humans to histocompatibility antigens (Class I and II) presented by frozen osteochondral allografts. These observations include a correlation of immune responses with long-term clinical outcome. As found in animal models, matching of histocompatibility antigens, particularly to Class II, improves clinical and, presumably, biologic success following implantation of massive frozen bone allografts in humans. The presence of sensitization clearly does not preclude a satisfactory outcome, nor have other reconstructive alternatives (e.g., metallic implants) been shown to be superior in their long-term results. PMID- 10471764 TI - Osteoconductive materials and their role as substitutes for autogenous bone grafts. AB - The term osteoconduction applies to a three-dimensional process that is observed when porous structures are implanted into or adjacent to bone. Capillaries, perivascular tissues, and osteoprogenitor cells migrate into porous spaces and incorporate the porous structure with newly formed bone. The observed process is characterized by an initial ingrowth of fibrovascular tissue that invades the porous structure followed by the later development of new bone applied directly within it. This article reviews observations of commonly used osteoconductive matrices to increase understanding of this process. PMID- 10471765 TI - Coralline bone graft substitutes. AB - Coralline porous ceramics are biocompatible and osteoconductive implants. They have proven to be effective as bone graft substitutes in large animal models and in humans. Bone and supporting soft tissue grow into and throughout their porosity if the implant is placed in direct apposition to viable bone and the interfaces are stabilized. The bone within the implant remodels in response to Wolff's law. Both the implant properties (chemistry and porosity) and the biologic environment modulate the rate of implant resorption. Composite technology with resorbable polymers can improve the mechanical properties of these ceramics. PMID- 10471766 TI - Calcium sulfate- and calcium phosphate-based bone substitutes. Mimicry of the mineral phase of bone. AB - Calcium sulfate and calcium phosphate have provided the orthopedic surgeon a viable alternative to autogenous bone grafting as either an osteoconductive bone void filler or a bone graft extender. These materials mimic the mineral phase of bone and are resorbed at a rate similar to the rate of bone formation. Thus, they are able to provide some structural support and prevent ingrowth of fibrous tissue while facilitating creeping substitution by the host bone. PMID- 10471767 TI - Gene therapy for osteoinduction. AB - One application of gene therapy that holds great promise is the stimulation of bone formation. Gene therapy offers several potential advantages over other methods of osteoinduction and current research suggests that it may be a feasible treatment option for the orthopedic surgeon in the near future. This article reviews the basic concepts and strategies of gene therapy and evaluates the current research using gene therapy to induce bone formation and enhance healing. PMID- 10471768 TI - Osteoinductive bone graft substitutes for spinal fusion: a basic science summary. AB - An understanding of precise biologic mechanisms at work during spinal fusion healing is just beginning to evolve. Current molecular biology research has shown this process to be multifactorial and extremely complex. With the advent of improved animal models to study the biology of spinal fusion, essential information regarding the basic science behind arthrodesis has advanced knowledge of this process. Moreover, with advances in local gene therapy as well as osteoinductive proteins and osteoinductive carrier matrices, the orthopedic surgeon will soon enter into a new era of biologic manipulation for fusion. PMID- 10471769 TI - Osteoinductive growth factors in preclinical fracture and long bone defects models. AB - Fracture healing is a specialized form of the reparative process that the musculoskeletal system undergoes to restore skeletal integrity. This biologic process is a consequence of a complex cascade of biologic events that result in the restoration of bone tissue, allowing for the resumption of musculoskeletal function. Several growth-promoting substances have been identified at the site of skeletal injury and appear to play a physiologic role in fracture healing. This article reviews the effects of these osteoinductive growth factors on bone healing as elucidated by both preclinical in vivo fracture and diaphyseal defect healing models. PMID- 10471770 TI - Bone grafts and bone substitutes in hip and knee surgery. AB - Bone grafts and bone substitutes are an essential part of the armamentarium of orthopedic surgeons. This article presents the current knowledge about bone replacements and reviews the available sources, techniques, and indications of the various types of autograft, allograft, and synthetic agents that are used in contemporary hip and knee replacement surgery. PMID- 10471771 TI - Tumor reconstruction. AB - This article discusses opinions for reconstruction of bony defects created by tumor resections. The literature is reviewed, and the results of different reconstruction methods from the Massachusetts General and Children's Hospitals' database are discussed. The authors' treatment goals and algorithms are reviewed as well as the rationale behind them. PMID- 10471772 TI - Bone grafting for spinal fusion. AB - At least 250,000 spinal fusions are performed in the United States each year, nearly all requiring implantation of bone graft material. The preferred technique for most of these operations is the transplantation of structured or morcellized autologous corticocancellous bone from the iliac crest. Further, because of the increasing frequency of spinal fusion surgery during the 1990s, arthrodesis of the spine has become the most common reason for autologous bone graft harvest. This article reviews the current clinical status of autogenous bone grafts and alternative materials in spinal fusion surgery. PMID- 10471773 TI - Inhibition of protein tyrosine phosphatases blocks calcium-induced activation of metaphase II-arrested oocytes of Xenopus laevis. AB - We have studied the effect of a protein tyrosine phosphatases (PTP) inhibitor on calcium-induced activation of Xenopus laevis oocytes arrested at metaphase II. Ammonium molybdate microinjection blocked pronucleus formation following A23187 treatment while cortical granules still underwent exocytosis. Pronuclei still occurred in ammonium molybdate-injected oocytes following 6-DMAP addition. Changes that usually occurred following A23187 exposure were inhibited in the presence of ammonium molybdate in the oocyte: MAPK dephosphorylation, p34(cdc2) rephosphorylation and cyclin B2 and p39(mos) proteolysis. These results suggest that a PTP is involved in the activation of the ubiquitin-dependent degradation machinery. PMID- 10471774 TI - Refsum disease diagnostic marker phytanic acid alters the physical state of membrane proteins of liver mitochondria. AB - Phytanic acid (3,7,11,15-tetramethylhexadecanoic acid), a branched chain fatty acid accumulating in Refsum disease to high levels throughout the body, induces uncoupling of rat liver mitochondria similar to non-branched fatty acids (e.g. palmitic acid), but the contribution of the ADP/ATP carrier or the aspartate/glutamate carrier in phytanic acid-induced uncoupling is of minor importance. Possible deleterious effects of phytanic acid on membrane-linked energy coupling processes were studied by ESR spectroscopy using rat liver mitochondria and a membrane preparation labeled with the lipid-specific spin probe 5-doxylstearic acid (5-DSA) or the protein-specific spin probe MAL-TEMPO (4 maleimido-2,2,6, 6-tetramethyl-piperidine-1-oxyl). The effects of phytanic acid on phospholipid molecular dynamics and on the physical state of membrane proteins were quantified by estimation of the order parameter or the ratio of the amplitudes of the weakly to strongly immobilized MAL-TEMPO binding sites (W/S ratio), respectively. It was found, that phytanic acid (1) increased the mobility of phospholipid molecules (indicated by a decrease in the order parameter) and (2) altered the conformational state and/or the segmental mobility of membrane proteins (indicated by a drastic decrease in the W/S ratio). Unsaturated fatty acids with multiple cis-double bonds (e.g. linolenic or arachidonic acid), but not non-branched FFA (ranging from chain length C10:0 to C18:0), also decrease the W/S ratio. It is hypothesized that the interaction of phytanic acid with transmembrane proteins might stimulate the proton permeability through the mitochondrial inner membrane according to a mechanism, different to a protein supported fatty acid cycling. PMID- 10471775 TI - Receptor-mediated activation of phospholipase D by sphingosine 1-phosphate in skeletal muscle C2C12 cells. A role for protein kinase C. AB - The present study showed that sphingosine 1-phosphate (SPP) induced rapid stimulation of phospholipase D (PLD) in skeletal muscle C2C12 cells. The effect was receptor-mediated since it was fully inhibited by pertussis toxin. All known SPP-specific receptors, Edg-1, Edg-3 and AGR16/H218, resulted to be expressed in C2C12 myoblasts, although at a different extent. SPP-induced PLD activation did not involve membrane translocation of PLD1 or PLD2 and appeared to be fully dependent on protein kinase C (PKC) catalytic activity. SPP increased membrane association of PKCalpha, PKCdelta and PKClambda, however, only PKCalpha and PKCdelta played a role in PLD activation since low concentrations of GF109203X and rottlerin, a selective inhibitor of PKCdelta, prevented PLD stimulation. PMID- 10471776 TI - Analysis of the domain structure of elongation factor-2 kinase by mutagenesis. AB - A number of elongation factor-2 kinase (eEF-2K) mutants were constructed to investigate features of this kinase that may be important in its activity. Typical protein kinases possess a highly conserved lysine residue in subdomain II which follows the GXGXXG motif of subdomain I. Mutation of two lysine residues, K340 and K346, which follow the GXGXXG motif in eEF-2K had no effect on activity, showing that such a lysine residue is not important in eEF-2K activity. Mutation of a conserved pair of cysteine residues C-terminal to the GXGXXG sequence, however, completely inactivated eEF-2K. The eEF-2K CaM binding domain was localised to residues 77-99 which reside N-terminal to the catalytic domain. Tryptophan 84 is an important residue within this domain as mutation of this residue completely abolishes CaM binding and eEF-2K activity. Removal of approximately 130 residues from the C-terminus of eEF-2K completely abolished autokinase activity; however, removal of only 19 residues inhibited eEF-2 kinase activity but not autokinase activity, suggesting that a short region at the C terminal end may be important in interacting with eEF-2. Likewise, removal of between 75 and 100 residues from the N-terminal end completely abolished eEF-2K activity. PMID- 10471777 TI - Enhancement of RNA self-cleavage by micellar catalysis. AB - It has been reported recently that naturally occurring catalytic RNAs like hammerhead and hairpin ribozyme do not require metal ions for efficient catalysis. It seems that the folded tertiary structure of the RNA contributes more to the catalytic function than was initially recognized. We found that a highly specific self-cleavage reaction can occur within a small bulge loop of four nucleotides in a mini-substrate derived from Arabidopsis thaliana intron containing pre-tRNA(Tyr) in the absence of metal ions. NH(4)(+) cations and non ionic or zwitter-ionic detergents at or above their critical micelle concentration are sufficient to catalyze this reaction. The dependence on micelles for the reaction leads to the assumption that physical properties, i.e. the hydrophobic interior of a micelle, are essential for this self-cleavage reaction. We suggest that NH(4)(+)-ions play a crucial role for the entry of the negatively charged RNA into the hydrophobic interior of a detergent micelle. A change of the pattern of hydration or hydrogen bonds caused by the hydrophobic surrounding enhances the reaction by a factor of 100. These findings suggest that highly structured RNAs may shift pK(a) values towards neutrality via the local environment and thereby enhance their ability to perform general acid-base catalysis without the participation of metal ions. PMID- 10471778 TI - Induction of myelin gene expression in Schwann cell cultures by an interleukin-6 receptor-interleukin-6 chimera. AB - Expression of myelin basic protein (MBP) and Po gene products is induced during the final postnatal maturation of Schwann cells and reinduced during nerve regeneration. We show that a chimeric protein containing interleukin-6 fused to its soluble receptor (IL6RIL6 chimera) induces MBP and Po RNAs and proteins in cultures of dorsal root ganglia (DRG) from 14 day old mouse embryos. Activation of gp130 signaling by IL6RIL6 appears comparable to cyclic AMP elevating agents to induce the myelin gene products in DRG and in pure Schwann cell cultures. PMID- 10471779 TI - Detergent-free membrane protein crystallization. AB - A comprehensive understanding of structure-function relationships of proteins requires their structures to be elucidated to high resolution. With most membrane proteins this has not been accomplished so far, mainly because of their notoriously poor crystallizability. Here we present a completely detergent-free procedure for the incorporation of a native purple membrane into a monoolein based lipidic cubic phase, and subsequent crystallization of three-dimensional bacteriorhodopsin crystals therein. These crystals exhibit comparable X-ray diffraction quality and mosaicity, and identical crystal habit and space group to those of bacteriorhodopsin crystals that are grown from detergent-solubilized protein in cubic phase. PMID- 10471780 TI - Overexpression of poly(A)-binding protein down-regulates the translation or the abundance of its own mRNA. AB - Poly(A)-binding protein (PABP) mRNA is subject to autoregulation through a 61 nucleotides long A-rich sequence in its 5' untranslated region (UTR). Here, we show that this mode of regulation is exerted in a cell type-specific manner. Thus, overexpression of PABP in mouse NIH 3T3 fibroblasts represses the translation of the respective endogenous mRNA or that of a chimeric mRNA containing just the 5' UTR of PABP mRNA. In contrast, ectopic expression of PABP in human embryonic kidney 293 cells down-regulates the abundance of the endogenous PABP mRNA, rather than affecting its translational efficiency. Transfection experiments with chimeric constructs suggest that the lack of translational autoregulation of endogenous PABP mRNA in these cells appears to reflect the presence of an overriding regulatory element outside the A-rich region. PMID- 10471781 TI - Coordinated regulation of ceruloplasmin and metallothionein mRNA by interleukin-1 and copper in HepG2 cells. AB - During the acute phase response, cytokines induce hepatic metallothionein and ceruloplasmin synthesis and the uptake of metals. We have investigated how copper and cytokines may interact in controlling ceruloplasmin (CP) and metallothionein mRNA in liver cells. We found that IL-1alpha, IL-1beta and IL-6 increased both metallothionein-1 (MT-1) and metallothionein-2 (MT-2) mRNA in HepG2 cells. The time and pattern of induction was different, both IL-1alpha and IL-1beta inducing two peaks of MT-1 and MT-2, with that of MT-2 being much larger. IL-6 induced only low levels of both MT-1 and MT-2 mRNA. CP mRNA was also increased after 16 h by IL-1beta, whereas IL-1alpha induced two CP peaks at 8 and 20 h, while IL-6 had little effect. Copper administration gave rise to substantially increased MT-1 mRNA, a slightly lower increase in MT-2 and also a significant increase in CP mRNA with similar kinetics. These parallel increases in MT and CP mRNA suggest that the coordinated expression of these proteins may be important for their synthesis during the acute phase response. PMID- 10471782 TI - Enhanced anti-HIV-1 activity of CC-chemokine LD78beta, a non-allelic variant of MIP-1alpha/LD78alpha. AB - We compared the anti-HIV-1 activity of CC-chemokine LD78beta with that of MIP 1alpha, another CC-chemokine which shows 94% sequence homology with LD78beta. Despite its close similarity to MIP-1alpha, the anti-HIV-1 activity of LD78beta appeared to be nearly 10 times higher than that of MIP-1alpha. Mutagenesis of MIP 1alpha showed that the N-terminal additional tetrapeptide, which was present in LD78beta and absent in MIP-1alpha, is responsible for enhanced anti-HIV-1 activity. The N-terminal structure-function relationship of LD78beta described here will be of value in understanding the chemokine-receptor interactions and designing anti-HIV-1 compounds based on LD78beta. PMID- 10471783 TI - Role of a bound ubiquinone on reactions of the Escherichia coli cytochrome bo with ubiquinol and dioxygen. AB - To probe the functional role of a bound ubiquinone-8 in cytochrome bo-type ubiquinol oxidase from Escherichia coli, we examined reactions with ubiquinol-1 and dioxygen. Stopped-flow studies showed that anaerobic reduction of the wild type and the bound ubiquinone-free (DeltaUbiA) enzymes with ubiquinol-1 immediately takes place with four kinetic phases. Replacement of the bound ubiquinone with 2,6-dibromo-4-cyanophenol (PC32) suppressed the anaerobic reduction of the hemes with ubiquinol-1 by eliminating the fast phase. Flow-flash studies in the reaction of the fully reduced enzyme with dioxygen showed that the heme b-to-heme o electron transfer occurs with a rate constant of approximately 1x10(4) s(-1) in all three preparations. These results support our previous proposal that the bound ubiquinone is involved in facile oxidation of substrates in subunit II and subsequent intramolecular electron transfer to low-spin heme b in subunit I. PMID- 10471784 TI - STABLE: protein-DNA fusion system for screening of combinatorial protein libraries in vitro. AB - We have developed a new method that permits the complete in vitro construction and selection of peptide or protein libraries. This method relies on an in vitro transcription/translation reaction compartmentalized in water in oil emulsions. In each emulsion compartment, streptavidin (STA)-fused polypeptides are synthesized and attached to the encoding DNA via its biotin label. The resulting protein-DNA fusion molecules recovered from the emulsion can be subjected to affinity selection based on the properties of the peptide portion, whose sequence can be determined from that of its DNA-tag. This method, named 'STABLE' (STA biotin linkage in emulsions), should be useful for rapid in vitro evolution of proteins and for ligand-based selection of cDNA libraries. PMID- 10471785 TI - An ABC transporter homologous to TAP proteins. AB - Polymerase chain reaction amplification of cDNA from rat intestine revealed the expression of a novel ABC transporter, TAPL (TAP-like). Subsequently, the protein sequence was deduced from the nucleotide sequence of cDNA carrying the entire coding region. TAPL is transcribed ubiquitously in various rat tissues. The protein, with 762 amino acid residues, has potential transmembrane domains, and an ATP-binding domain in its amino and carboxyl terminal regions, respectively, and is highly homologous to TAP1 and TAP2 (transporters associated with antigen presentation/processing): pairwise comparisons with TAPL demonstrated 39 and 41% of the residues are identical, respectively. These numerical values are essentially the same as that for TAP1 and TAP2 (39%), and the hydropathy profiles of TAPL, TAP1 and TAP2 are quite similar. The similarity among these three proteins suggests that they could be derived from a common ancestral gene. Furthermore, we found that there is a potential splicing isoform, sharing the amino terminal 720 amino acid residues of TAPL. PMID- 10471786 TI - Bromine K-edge EXAFS studies of bromide binding to bromoperoxidase from Ascophyllum nodosum. AB - Bromine K-edge EXAFS studies have been carried out for bromide/peroxidase samples in Tris buffer at pH 8. The results are compared with those of aqueous (Tris buffered) bromide and vanadium model compounds containing Br-V, Br-C(aliphatic) and Br-C(aromatic) bonds. It is found that bromide does not coordinate to the vanadium centre. Rather, bromine binds covalently to carbon. A possible candidate is active site serine. PMID- 10471787 TI - Lysophosphatidylcholine phosphorylates CREB and activates the jun2TRE site of c jun promoter in vascular endothelial cells. AB - Lysophosphatidylcholine (lyso-PC), a polar phospholipid increased in atherogenic lipoproteins and atherosclerotic lesions, has been shown to induce transcription of a variety of endothelial genes relevant to atherogenesis. Lyso-PC has been shown to activate c-jun N-terminal kinase (JNK) and activator protein 1 (AP-1) and thereby stimulate transcription of the c-jun gene. Here we provide evidence that lyso-PC can phosphorylate cyclic AMP responsive element binding protein (CREB) and thereby activate the jun2 12-O-tetradecanoylphorbol 13-acetate response element (jun2TRE) site of the c-jun promoter, which appears to be the major molecular mechanism involved in lyso-PC-induced c-jun gene expression in cultured bovine aortic endothelial cells (BAEC). Transient transfection of BAEC with a 1.6-kbp c-jun promoter and luciferase reporter fusion gene resulted in a 12.9-fold increase in luciferase activity by lyso-PC treatment. Serial deletion mutation in c-jun promoter and luciferase reporter gene assay revealed that the 5' promoter region between nucleotide numbers -268 and -127, which contains a jun2TRE binding sequence, was most crucial for lyso-PC-induced transcription. The 5' promoter region between -76 and -27, which contains an AP-1 site, also affected lyso-PC-induced transcription of the c-jun gene. Point mutation in the jun2TRE site reduced lyso-PC-induced transcription of the c-jun promoter luciferase fusion gene by a 70.3% decrease in c-jun promoter activity. Electrophoretic mobility shift assays showed increased binding of (32)P-labeled oligonucleotides with jun2TRE in nuclear extracts isolated from lyso-PC-treated BAEC, which was abolished or supershifted by anti-CREB antibody. Immunoblotting with anti-phosphorylated CREB antibody showed rapid phosphorylation of this protein after lyso-PC treatment. These results indicate that lyso-PC phosphorylates CREB, which was then bound to the jun2TRE site of the c-jun promoter and activated transcription. Activation of jun2TRE may play a key role in the transcriptional activation of c-jun as well as other endothelial genes depending upon these transcription factors. PMID- 10471788 TI - R-esp1, a rat homologue of drosophila groucho, is differentially expressed after optic nerve crush and mediates NGF-induced survival of PC12 cells. AB - The differential display reverse transcription polymerase chain reaction method was used to detect alterations in gene expression in the superior colliculus after optic nerve crush in adult rats. One of the most prominent changes observed was the selective induction of R-esp1, a homologue of the Drosophila enhancer of split locus (Groucho). Therefore, we studied the influence of R-esp1 on nerve growth factor (NGF)-induced cell survival of PC12 cells. Overexpression of R-esp1 promotes cell survival even in the absence of NGF and, conversely, it is reduced by antisense-mediated inhibition of R-esp1 expression. In conclusion, we propose a novel model in which R-esp1 protein mediates the NGF-signaling pathway. PMID- 10471789 TI - Association of the rat heterogeneous nuclear RNA-ribonucleoprotein F with TATA binding protein. AB - Heterogeneous nuclear ribonucleoprotein F (hnRNP-F) has been shown to be a pre mRNA splicing factor. Recent studies have uncovered the coordination of synthesis of pre-mRNA and its processing, including post-transcriptional modification and splicing. Here, we present evidence for an association between a splicing factor, hnRNP-F, and TATA-binding protein (TBP), which is an essential factor needed for transcription initiation. An affinity detection experiment revealed hnRNP-F in the preparation of TBP-interacting proteins. HnRNP-F was associated with TBP in nuclear extracts and was capable of direct binding to TBP in vitro. These results suggest that hnRNP-F is associated with TBP in the cell. HnRNP-F was observed in abundance in the thymus, spleen and testis, and its distribution pattern was similar to that of TBP, implying a functional coordination of transcription and splicing. We assume that the splicing machinery is associated with the transcription apparatus as a prerequisite prior to transcriptional elongation. PMID- 10471790 TI - Involvement of branched-chain amino acid aminotransferase (Bcat1/Eca39) in apoptosis. AB - The branched-chain amino acid aminotransferase, Bcat1/Eca39, catalyzes the first step of branched-chain amino acid catabolism. Bcat1/Eca39 was originally isolated from a c-myc-induced tumor and was proven to be a direct target for c-Myc regulation. The gene is highly conserved in evolution and disruption of its yeast homolog affects cell growth. To assess the role of Bcat1/Eca39 in mammalian cells, we overexpressed Bcat1/Eca39 in murine cells and studied effects on cell growth. Overexpression of Bcat1/Eca39 had no apparent effect on the proliferation of cells grown with high serum concentrations, but under serum deprivation conditions, led to a decrease in cell viability. Cell death under these conditions displayed apoptotic features. The branched-chain keto acid, alpha ketoisocaproate, a metabolite of leucine catabolism produced by BCAT1/ECA39, was previously found to inhibit cell growth. We show that alpha-ketoisocaproate can induce rapid apoptotic cell death. This observation suggests that the growth inhibitory effect of BCAT1/ECA39 and its apoptosis promoting effect may be mediated by the levels of the products of BCAT1/ECA39 activity, namely, branched chain keto acids. PMID- 10471791 TI - CA-MMP: a matrix metalloproteinase with a novel cysteine array, but without the classic cysteine switch. AB - A matrix metalloproteinase (MMP)-like gene was identified in mouse to contain a conserved MMP catalytic domain and an RRRR motif. It lacks a classic cysteine switch, but it possesses two novel motifs: a cysteine array (Cys-X(6)-Cys-X(8) Cys-X(10)-Cys-X(3)-Cys-X(2)-Cys), and a novel Ig-fold. It is named CA-MMP after the distinct cysteine array motif, and little is known about its biochemical function. In an attempt to characterize CA-MMP activity, the full-length sequence was expressed in mammalian cells and its product found to be cell-associated without detectable secretion. In light of this unusual finding, a chimera combining the catalytic domain of CA-MMP with the prodomain of stromelysin-3 was constructed to express a fully active enzyme in mammalian cells. Purified CA-MMP catalytic domain expresses proteolytic activity against protein substrates in an MMP inhibitor sensitive fashion. Taken together, it is concluded that CA-MMP is an MMP with distinct structure, biochemical properties and evolutionary history that may define a new subclass of the MMP superfamily. PMID- 10471792 TI - Interleukin 2-Bax: a novel prototype of human chimeric proteins for targeted therapy. AB - During the past few years many chimeric proteins have been developed to target and kill cells expressing specific surface molecules. Generally, these molecules carry a bacterial or plant toxin that destroys the unwanted cells. The major obstacle in the clinical application of such chimeras is their immunogenicity and non-specific toxicity. We have developed a new generation of chimeric proteins, taking advantage of apoptosis-inducing proteins, such as the human Bax protein, as novel killing components. The first prototype chimeric protein, IL2-Bax, directed toward IL2R-expressing cells, was constructed, expressed in Escherichia coli and partially purified. IL2-Bax increased the population of apoptotic cells in a variety of target T cell lines, as well as in human fresh PHA-activated lymphocytes, in a dose-dependent manner and had no effect on cells lacking IL2R expression. The IL2-Bax chimera represents an innovative approach for constructing chimeric proteins comprising a molecule that binds a specific cell type and an apoptosis-inducing protein. Such new chimeric proteins could be used for targeted treatment of human diseases. PMID- 10471793 TI - Photoinduced electron transfer in singly labeled thiouredopyrenetrisulfonate azurin derivatives. AB - A novel method for the initiation of intramolecular electron transfer reactions in azurin is reported. The method is based on laser photoexcitation of covalently attached thiouredopyrenetrisulfonate (TUPS), the reaction that generates the low potential triplet state of the dye with high quantum efficiency. TUPS derivatives of azurin, singly labeled at specific lysine residues, were prepared and purified to homogeneity by ion exchange HPLC. Transient absorption spectroscopy was used to directly monitor the rates of the electron transfer reaction from the photoexcited triplet state of TUPS to Cu(II) and the back reaction from Cu(I) to the oxidized dye. For all singly labeled derivatives, the rate constants of copper ion reduction were one or two orders of magnitude larger than for its reoxidation, consistent with the larger thermodynamic driving force for the former process. Using 3-D coordinates of the crystal structure of Pseudomonas aeruginosa azurin and molecular structure calculation of the TUPS modified proteins, electron transfer pathways were calculated. Analysis of the results revealed a good correlation between separation distance from donor to Cu ligating atom (His-N or Cys-S) and the observed rate constants of Cu(II) reduction. PMID- 10471794 TI - Circularly permuted variants of the green fluorescent protein. AB - Folding of the green fluorescent protein (GFP) from Aequorea victoria is characterized by autocatalytic formation of its p-hydroxybenzylideneimidazolidone chromophore, which is located in the center of an 11-stranded beta-barrel. We have analyzed the in vivo folding of 20 circularly permuted variants of GFP and find a relatively low tolerance towards disruption of the polypeptide chain by introduction of new termini. All permuted variants with termini in strands of the beta-barrel and about half of the variants with termini in loops lost the ability to form the chromophore. The thermal stability of the permuted GFPs with intact chromophore is very similar to that of the wild-type, indicating that chromophore side chain interactions strongly contribute to the extraordinary stability of GFP. PMID- 10471795 TI - Novel reactions involved in energy conservation by methanogenic archaea. AB - Methanogenic archaea of the order Methanosarcinales which utilize C(1) compounds such as methanol, methylamines or H(2)+CO(2), employ two novel membrane-bound electron transport systems generating an electrochemical proton gradient: the H(2):heterodisulfide oxidoreductase and the F(420)H(2):heterodisulfide oxidoreductase. The systems are composed of the heterodisulfide reductase and either a membrane-bound hydrogenase or a F(420)H(2) dehydrogenase which is functionally homologous to the proton-translocating NADH dehydrogenase. Cytochromes and the novel electron carrier methanophenazine are also involved. In addition, the methyl-H(4)MPT:HS-CoM methyltransferase is bioenergetically relevant. The enzyme couples methyl group transfer with the translocation of sodium ions and seems to be present in all methanogens. The proton-translocating systems with the participation of cytochromes and methanophenazine have been found so far only in the Methanosarcinales. PMID- 10471796 TI - Interplay of organic and biological chemistry in understanding coenzyme mechanisms: example of thiamin diphosphate-dependent decarboxylations of 2-oxo acids. AB - With the publication of the three-dimensional structures of several thiamin diphosphate-dependent enzymes, the chemical mechanism of their non-oxidative and oxidative decarboxylation reactions is better understood. Chemical models for these reactions serve a useful purpose to help evaluate the additional catalytic rate acceleration provided by the protein component. The ability to generate, and spectroscopically observe, the two key zwitterionic intermediates invoked in such reactions allowed progress to be made in elucidating the rates and mechanisms of the elementary steps leading to and from these intermediates. The need remains to develop chemical models, which accurately reflect the enzyme-bound conformation of this coenzyme. PMID- 10471797 TI - Constitutively active mutants of the beta1-adrenergic receptor. AB - We provide the first evidence that point mutations can constitutively activate the beta(1)-adrenergic receptor (AR). Leucine 322 of the beta(1)-AR in the C terminal portion of its third intracellular loop was replaced with seven amino acids (I, T, E, F, C, A and K) differing in their physico-chemical properties. The beta(1)-AR mutants expressed in HEK-293 cells displayed various levels of constitutive activity which could be partially inhibited by some beta-blockers. The results of this study might have interesting implications for future studies aiming at elucidating the activation process of the beta(1)-AR as well as the mechanism of action of beta-blockers. PMID- 10471798 TI - The 25 kDa protein recognizing the rat curved region upstream of the origin of the L-strand replication is the rat homologue of the human mitochondrial transcription factor A. AB - Mass spectrometry matrix-assisted laser desorption ionization (MALDI) analysis and N-terminus sequencing as well as immunoblotting experiments using human and mouse antibodies have allowed us to identify the 25 kDa protein, previously isolated from rat liver using magnetic beads coated with a rat liver mitochondrial (mt) DNA region upstream of the Ori-L, as the homologue of human mt transcription factor A (mtTFA). We can therefore identify this DNA binding protein as the rat mtTFA. Furthermore, since we previously showed that the 25 kDa protein purified from rat liver was able to bind the curved mtDNA region upstream of the Ori-L as well as the curved mtDNA in the D-loop region, the results here reported lead us to state, for the first time, that mtTFA binds both the curved regions of mtDNA upstream of the two replication origins. PMID- 10471799 TI - Different cation binding to the I domains of alpha1 and alpha2 integrins: implication of the binding site structure. AB - In the present work, we studied the interactions of recombinant alpha1 and alpha2 integrin I domains with cations Tb(3+), Mn(2+), Mg(2+) and Ca(2+). We observed that alpha1 and alpha2 I domains bind these cations with significantly different characteristics. The binding of Mg(2+) by the alpha1 I domain was accompanied by significant changes of tryptophan fluorescence which could be interpreted as a conformational change. Comparison of the alpha1 integrin I domain structure obtained by comparative modeling with a known structure of the alpha2 integrin I domain shows distinct differences in the metal ion binding sites which could explain the differences in cation binding. PMID- 10471800 TI - Oligomeric complexes involved in translocation of proteins across the membrane of the endoplasmic reticulum. AB - Proteins involved in protein translocation across the membrane of the endoplasmic reticulum assemble into different oligomeric complexes depending on their state of function. To analyse such membrane protein complexes we fractionated proteins of mammalian rough microsomes and analysed them using blue native PAGE and immunoblotting. Among the proteins characterised are the Sec61p complex, the oligosaccharyl transferase (OST) complex, the translocon-associated protein (TRAP) complex, the TRAM and RAMP4 proteins, the signal recognition particle (SRP) and the SRP receptor (SR). Interestingly, the RAMP4 protein, SR and OST complex display more than one oligomeric form. PMID- 10471801 TI - Kinetics of CheY phosphorylation by small molecule phosphodonors. AB - The chemotaxis response regulator CheY can acquire phosphoryl groups either from its associated autophosphorylating protein kinase, CheA, or from small phosphodonor molecules such as acetyl phosphate. We report a stopped-flow kinetic analysis of CheY phosphorylation by acetyl phosphate. The results show that CheY has a very low affinity for this phosphodonor (K(s)&z.Gt;0.1 M), consistent with the conclusion that, whereas CheY provides catalytic functions for the phosphotransfer reaction, the CheA kinase may act simply to increase the effective phosphodonor concentration at the CheY active site. PMID- 10471802 TI - Comparison of DeltapH- and Delta***φ***-driven ATP synthesis catalyzed by the H(+)-ATPases from Escherichia coli or chloroplasts reconstituted into liposomes. AB - The H(+)-ATPases from Escherichia coli, EF(0)F(1), and from chloroplasts, CF(0)F(1), were reconstituted in liposomes from phosphatidylcholine/phosphatidic acid. The proteoliposomes were energized by an acid-base transition and a K(+)/valinomycin diffusion potential and the initial rate of ATP synthesis was measured as a function of the transmembrane pH difference, DeltapH, and the electric potential difference, Deltaφ. With EF(0)F(1), a rate of 80 s(-1) is observed at DeltapH=4.1 and Deltaφ approximately 140 mV. The rate decreases sigmoidally with Deltaφ and at Deltaφ approximately 0 mV, the rate is about 1 s(-1) although DeltapH is still 4.1. Under the same conditions with CF(0)F(1), a rate of 280 s(-1) is observed which decreases to 190 s(-1) when Deltaφ is abolished, i.e. ATP synthesis catalyzed by EF(0)F(1) and CF(0)F(1) depends in a different way on DeltapH and Deltaφ. EF(0)F(1)-catalyzed ATP synthesis was measured as a function of DeltapH at a constant Deltaφ. The rate depends sigmoidally on DeltapH reaching a maximal rate which cannot be further increased by increasing DeltapH. However, this maximal rate depends on Deltaφ, i.e. DeltapH and Deltaφ are not kinetically equivalent in driving ATP synthesis. We assume that EF(0)F(1) must be converted into a metastable, active state before it catalyzes proton transport-coupled ATP synthesis. For EF(0)F(1), this activation step depends only on Deltaφ, whereas for CF(0)F(1), the activation depends on DeltapH and Deltaφ. PMID- 10471803 TI - The proteasome inhibitor PI31 competes with PA28 for binding to 20S proteasomes. AB - PI31 is a previously described inhibitor of 20S proteasomes. Using recombinant PI31 we have analyzed its effect on proteasomal hydrolyzing activity of short fluorogenic substrates and of a synthetic 40-mer polypeptide. In addition, we investigated its influence on the activation of 20S proteasome by the proteasome activator PA28. PI31 inhibits polypeptide degradation already at concentrations which only partially inhibit fluorogenic substrate turnover and immunosubunits do not influence the PI31 binding affinity. Furthermore our data demonstrate that PI31 is a potent competitor of PA28-mediated activation. PMID- 10471804 TI - Alternative exon 3 splicing of the human major protein zero gene in white blood cells and peripheral nerve tissue. AB - The major protein zero (MPZ) is involved in peripheral myelin folding. Using nested reverse transcription-PCR, we amplified several fragments of MPZ mRNAs in white blood cells and in peripheral nerve tissue. Cloning of PCR products revealed the existence of three alternative splicing patterns: one resulted in the complete loss of exon 3 and two others induced partial skipping of the exon 3 sequence. All three alternative splicing mechanisms produced a frame-shift and created an identical premature stop codon in exon 4. We conclude that the existence of these MPZ RNA transcript variants may be the result of deliberate splicing decisions and may have functional implications in the cell. PMID- 10471805 TI - The beta-isoform of heat shock protein hsp-90 is structurally related with human microtubule-interacting protein Mip-90. AB - Through major research advances in the study of cytoskeletal organization, an integrated view of the complexity of this system has emerged. Recent findings on the microtubule-interacting protein Mip-90, which associates with microtubules and actin filaments in different cell domains, have shed light on its roles in cytoskeletal regulation. In order to study structural features of Mip-90, we sequenced several peptide fragments. A comparative sequence analysis revealed a high degree of similarity between the primary structure of this protein and the human heat shock protein of 90 kDa (hsp-90). Taken together, the present studies indicate the identity between Mip-90 and the the beta-isoform of hsp-90 (hsp 90beta). Western blot assays with an anti-hsp-90 monoclonal antibody showed cross reactivity of hsp-90 and Mip-90 affinity purified from HeLa cells. Furthermore, the observed structural identity of Mip-90 with the hsp-90beta was sustained by immunoblot assays using monoclonal antibodies that specifically recognize the alpha- and beta-forms of hsp-90. Comparative fingerprinting analysis, along with the evidence of a remarkably similar biochemical behavior of both hsp-90 and Mip 90 in different affinity chromatographic systems, supported these observations. These studies, along with previous investigations, provide new data to elucidate the functional significance of these interesting cellular components and its relationships with other proteins linked to the cell architecture. PMID- 10471806 TI - Voltammetric detection of superoxide production by photosystem II. AB - Oxygen radicals play both pathological and physiological roles in biological systems. The detection of such radicals is difficult due to their transient nature and the presence of highly efficient antioxidant mechanisms. In plants the physiological role of oxygen is twofold, oxygen is produced by the oxidation of water and consumed as an electron acceptor. The direct involvement of oxygen in photosynthetic events exposes the photosynthetic apparatus to a high probability of damage by oxygen radicals. We report here a direct, simple and rapid method for the measurement of superoxide in vitro based on voltammetric detection. It has potential applications for other in vitro systems investigating superoxide production. We show that in addition to the well established production of superoxide from photosystem I, under reducing conditions superoxide is also produced by photosystem II, probably from the Q(A) site. PMID- 10471807 TI - Human membrane type-4 matrix metalloproteinase (MT4-MMP) is encoded by a novel major transcript: isolation of complementary DNA clones for human and mouse mt4 mmp transcripts. AB - Five distinct membrane-type matrix metalloproteinases (MT-MMP) have been reported by cDNA cloning. However, the mt4-mmp gene product (MMP-17) has not been identified yet in spite of the cDNA isolation [Puente et al. (1996), Cancer Res. 56, 944-949]. In this study, we re-examined the transcripts for human mt4-mmp by 5' RACE and identified two types of transcripts. The minor one corresponded to the cDNA reported by Puente et al. and failed to express protein, and the other is the major transcript that has an extended open reading frame and expressed 67 and 71 kDa translation products. Thus, functional mt4-mmp has been identified for the first time. PMID- 10471808 TI - Essential arginine residues in maize starch synthase IIa are involved in both ADP glucose and primer binding. AB - The arginine-specific reagent phenylglyoxal inactivated the activity of maize starch synthase IIa (SSIIa), due to the modification of at least one arginine residue out of a possible 42. The addition of ADPGlc completely protected SSIIa from the inactivation, indicating that arginine may be involved in the interaction of this anionic substrate with SSIIa. However, site-directed mutagenesis of the conserved Arg-214 in SSIIa showed that this amino acid is important for apparent affinity of SSIIa for its primer (amylopectin and glycogen), as evidenced by a marked increase in the K(m) for primer upon substitution of this amino acid with no concomitant change in V(max), K(m) for ADPGlc, or secondary structure. Therefore, Arg-214 of SSIIa appears to play a role in its primer binding. PMID- 10471809 TI - Tpr1, a Schizosaccharomyces pombe protein involved in potassium transport. AB - The Schizosaccharomyces pombe Tpr1 was isolated as suppressor of the Saccharomyces cerevisiae Delta trk1,2 potassium uptake deficient phenotype. Tpr1, for tetratrico peptide repeat, encodes a 1039 amino acid residues protein with several reiterated TPR units displaying significant homology to p150(TSP), a recently identified phosphoprotein of mouse, to S. cerevisiae CTR9 and to related sequences of human, Caenorhabditis elegans, Methanoccocus jannaschii and Arabidopsis thaliana. Expression of Tpr1 restored growth on 0.2 mM K(+) media, induced K(+) transport with a K(T) of 4.6 mM and resumed inward currents of -90 pA at -250 mV (pH 7.2) conducting K(+) and other alkali-metal ions. The tetratrico peptide repeat is a degenerate motif of 34 amino acids that is repeated several times within TPR-containing proteins and has been suggested to mediate protein-protein interactions. The sequence and putative binding properties of Tpr1 suggest the protein unlikely as transporter but involved in the enhancement of K(+) uptake via conventional carriers. PMID- 10471810 TI - Localization of calponin binding sites in the structure of 90 kDa heat shock protein (Hsp90). AB - The structure of rabbit liver Hsp90 was reevaluated by limited trypsinolysis, N terminal sequencing and determination of the site that is phosphorylated by casein kinase II. Limited proteolysis results in formation of four groups of large peptides with M(r) in the range of 26-41 kDa. Peptides with M(r) 39-41 kDa were represented by large N-terminal and central peptides starting at residue 283 of the alpha-isoform of Hsp90. All sites phosphorylated by casein kinase II were located in the large 39-41 kDa peptides. Peptides with M(r) 26-27 kDa were represented by short N-terminal and central peptides starting at Glu-400 of the alpha-isoform of Hsp90. The data of affinity chromatography and light scattering indicate that smooth muscle calponin interacts with Hsp90. The calponin binding sites are located in the large (37-41 kDa) N-terminal and in a short (26-27 kDa) central peptide starting at Glu-400 of the alpha-isoform of Hsp90. Phosphorylation by casein kinase II up to 2 mol of phosphate per mol of Hsp90 does not affect interaction of Hsp90 with calponin. PMID- 10471811 TI - Sphingosylphosphorylcholine induces cytosolic Ca(2+) elevation in endothelial cells in situ and causes endothelium-dependent relaxation through nitric oxide production in bovine coronary artery. AB - Sphingosylphosphorylcholine (SPC) increased intracellular Ca(2+) concentration ([Ca(2+)]i) and nitric oxide (NO) production in endothelial cells in situ on bovine aortic valves, and induced endothelium-dependent relaxation of bovine coronary arteries precontracted with U-46619. The SPC-induced vasorelaxation was inhibited by N(omega)-monomethyl-L-arginine, an inhibitor of both constitutive and inducible NO synthase (NOS), but not by 1-(2-trifluoromethylphenyl) imidazole, an inhibitor of inducible NOS (iNOS). Immunoblotting revealed that endothelial constitutive NOS, but not iNOS, was present in endothelial cells in situ on the bovine aortic valves. We propose that SPC activates [Ca(2+)]i levels and NO production of endothelial cells in situ, thereby causing an endothelium dependent vasorelaxation. PMID- 10471812 TI - Expression of endothelin-1 in macrophages and mast cells in hyperplastic human tonsils. AB - In situ hybridization and immunohistochemical examinations of surgically resected specimens of human hyperplastic tonsils showed that macrophages in the germinal center and mast cells in the parafollicular and interfollicular areas expressed the transcript and protein of endothelin (ET)-1, but not ET-2 and ET-3. The macrophages appeared to be activated, since they possessed significant amounts of inducible nitric oxide synthase. None of these expressions was observed in normal tonsil. Our results suggest that the over-production of ET-1 by macrophages and mast cells may be involved in the pathogenesis of hyperplastic tonsils. PMID- 10471813 TI - A novel Jun N-terminal kinase (JNK)-binding protein that enhances the activation of JNK by MEK kinase 1 and TGF-beta-activated kinase 1. AB - We have identified a novel Jun N-terminal kinase (JNK)-binding protein, termed JNKBP1, and examined its binding affinity for JNK1, JNK2, JNK3, and extracellular signal-regulated kinase 2 (ERK2) in COS-7 cells. JNKBP1 preferentially interacted with the JNKs, but not with ERK2. Furthermore, we investigated the effect of overexpressing JNKBP1 on the JNK and ERK signaling pathways in COS-7 cells. JNKBP1 alone had only a marginal effect on JNK activity. However, the activation of JNK by MEK kinase 1 and TGF-beta-activated kinase 1 was significantly enhanced in the presence of JNKBP1. In contrast, JNKBP1 had no or very little effect on the ERK signaling pathway. These results suggest that JNKBP1 functions to facilitate the specific and efficient activation of the JNK signaling pathways. PMID- 10471814 TI - Differential discrimination of G-protein coupling of serotonin(1A) receptors from bovine hippocampus by an agonist and an antagonist. AB - We have studied the effect of guanosine-5'-O-(3-thiotriphosphate) (GTP-gamma-S), a non-hydrolyzable analogue of GTP, on agonist and antagonist binding to bovine hippocampal 5-hydroxytryptamine (5-HT)(1A) receptor in native membranes. Our results show that the specific binding of the agonist is inhibited with increasing concentrations of GTP-gamma-S along with a reduction in binding affinity. In sharp contrast to this, antagonist binding to 5-HT(1A) receptor shows no significant reduction and remains invariant over a large range of GTP gamma-S concentrations. The binding affinity of the antagonist also remains unaltered. This shows that the agonist and the antagonist differentially discriminate G-protein coupling of 5-HT(1A) receptors from bovine hippocampus. PMID- 10471815 TI - High level expression of Thermococcus litoralis 4-alpha-glucanotransferase in a soluble form in Escherichia coli with a novel expression system involving minor arginine tRNAs and GroELS. AB - The Thermococcus litoralis 4-alpha-glucanotransferase (GTase) gene has a high content of AGA and AGG codons for arginine, which are extremely rare in Escherichia coli. Expression of the GTase gene in E. coli resulted in low protein production and the accumulation of inclusion bodies. However, simultaneous expression of GTase with tRNA(AGA), tRNA(AGG) and GroELS affected both the production and solubility of GTase, and production of soluble GTase increasing about 5-fold. This new E. coli expression system should be applicable to the expression of not only archaeal but also eukaryotic genes, which usually contain a large number of AGA and AGG codons. PMID- 10471816 TI - The peroxisomal membrane protein Pex14p of Hansenula polymorpha is phosphorylated in vivo. AB - Hansenula polymorpha Pex14p (HpPex14p) is a component of the peroxisomal membrane essential for peroxisome biogenesis. Here, we show that HpPex14p is phosphorylated in vivo. In wild-type H. polymorpha cells, grown in the presence of [32P]orthophosphate, the 32P label was incorporated into HpPex14p. Labelled HpPex14p was induced after a shift of cells to methanol-containing media and rapidly disappeared after a shift to glucose medium, which induces specific peroxisome degradation. Alkaline phosphatase treatment of labelled HpPex14p resulted in the release of 32P and a minor shift of the HpPex14p band on Western blots. Phosphoamino acid analysis by two dimensional silica gel thin layer chromatography suggested that the major phosphoamino acid in phosphorylated HpPex14p was acid-labile. PMID- 10471817 TI - Molecular cloning of newt sex pheromone precursor cDNAs: evidence for the existence of species-specific forms of pheromones. AB - Cloning of cDNA encoding a decapeptide pheromone (sodefrin) that attracts conspecific female newts was attempted. A cDNA clone encoding a protein consisting of 189 amino acid residues including a sodefrin sequence was isolated from a Cynops pyrrhogaster abdominal gland cDNA library. Likewise, a cDNA clone encoding a molecule comparable to the sodefrin precursor was obtained from a Cynops ensicauda abdominal gland cDNA library. This clone encoded a precursor protein of 192 amino acid residues, including a sodefrin-like peptide sequence with substitutions of two amino acid residues. This is the first report of a peptide pheromone precursor in vertebrates. PMID- 10471818 TI - Induction of apoptosis of monocyte-macrophage lineage cells by 5-S-GAD. AB - We found that 5-S-GAD, an insect-derived antibacterial peptide, inhibited murine osteoclast formation in vitro. We examined the specific time point of the inhibitory action of 5-S-GAD on osteoclast formation and found that it mainly suppressed differentiation of osteoclasts in the middle of the culture period. Using HL60 cells that are able to differentiate into multinucleated macrophage like cells, we found that 5-S-GAD induced apoptosis of HL60 cells by producing H(2)O(2). Thus, the inhibition of osteoclast formation by 5-S-GAD could be, in part, due to apoptosis of the cells of an osteoclast lineage. PMID- 10471819 TI - Wide distribution of short interspersed elements among eukaryotic genomes. AB - Most short interspersed elements (SINEs) in eukaryotic genomes originate from tRNA and have internal promoters for RNA polymerase III. The promoter contains two boxes (A and B) spaced by approximately 33 bp. We used oligonucleotide primers specific to these boxes to detect SINEs in the genomic DNA by polymerase chain reaction (PCR). Appropriate DNA fragments were revealed by PCR in 30 out of 35 eukaryotic species suggesting the wide distribution of SINEs. The PCR products were used for hybridization screening of genomic libraries which resulted in identification of four novel SINE families. The application of this approach is illustrated by discovery of a SINE family in the genome of the bat Myotis daubentoni. Members of this SINE family termed VES have an additional B-like box, a putative polyadenylation signal and RNA polymerase III terminator. PMID- 10471820 TI - Long-range electrostatic effects on peptide folding. AB - The reversible folding/unfolding of a short peptide in solution is studied by molecular dynamics simulations. The effects of long-range electrostatic interactions are examined and found to be important both for the equilibrium between folded and unfolded states and the dynamics of the folding process. The neglect of long-range electrostatics leads to an increased population of unfolded states and increased structural fluctuations. When such interactions are taken into account, the peptide unfolds and folds to the experimentally determined structure several times during a 25 ns trajectory, with approximately equal populations of folded and unfolded states in the neighborhood of its proposed melting temperature. The effect of using spherical boundary conditions rather than periodic ones does not appear to have any major effect on the folding dynamics. PMID- 10471821 TI - Chiral DNA packaging in DNA-cationic liposome assemblies. AB - Recent studies have indicated that the structural features of DNA-lipid assemblies, dictated by the lipid composition and cationic lipid-to-DNA ratio, critically affect the efficiency of these complexes in acting as vehicles for cellular delivery of genetic material. Using circular dichroism we find that upon binding DNA, positively-charged liposomes induce a secondary conformational transition of the DNA molecules from the native B form to the C motif. Liposomes composed of positively-charged and neutral 'helper' lipids, found to be particularly effective as transfecting agents, induce - in addition to secondary conformational changes - DNA condensation into a left-handed cholesteric-like phase. A structural model is presented according to which two distinct, yet inter related modes of DNA packaging coexist within such assemblies. The results underline the notion that subtle changes in the components of a supramolecular assembly may substantially modulate the interplay of interactions which dictate its structure and functional properties. PMID- 10471822 TI - Increased expression of the Na(+)/Ca(2+) exchanger in the rat heart after immobilization stress is not induced by cortisol. AB - Calcium homeostasis is crucial for the proper function of cardiac cells. Since the Na(+)/Ca(2+) exchanger is an important modulator of calcium homeostasis especially in the heart, the objective of this study was to investigate the effect of immobilization stress on the high capacity Na(+)/Ca(2+) exchanger in rat heart ventricles and atria. Repeated immobilization stress increased both the mRNA and the protein level and the activity of the Na(+)/Ca(2+) exchanger in the left, but not the right ventricle of rat heart. Since corticosterone is rapidly increased during the stress stimulus, it might be assumed that mRNA of the Na(+)/Ca(2+) exchanger is increased through a glucocorticoid responsive element. However, we have found that cortisol did not change the Na(+)/Ca(2+) exchanger at the mRNA or protein levels. These results clearly show that this effect of stress is not mediated via cortisol. PMID- 10471823 TI - A novel downstream positive regulatory element mediating transcription of the human high mobility group (HMG) I-C gene. AB - The high mobility group (HMG) I proteins are small, non-histone chromosomal proteins that promote gene activation during development and within rapidly dividing cells. They do so by facilitating enhanceosome formation on inducible genes, via both protein/DNA and protein/protein interactions. The HMG I-C gene is tightly regulated, normally being expressed exclusively during embryonic development. However, HMG I-C expression is also observed frequently in a number of tumor types, and this expression has been shown to contribute to the malignant transformation process. With the aim of dissecting pathways that lead to aberrant expression of HMG I-C in tumor cells, we have analyzed HMG I-C gene regulation in the human hepatoma cell line PLC/PRF/5. One of the two HMG I-C transcripts detected in this cell line originates from a novel downstream initiation site at nucleotide -161 relative to the first methionine. Transcription from the downstream initiation site is mediated by a PRE located between nt -222 and -217. We show here that the Sp1 and Sp3 transcription factors interact with the PRE and transactivate the HMG I-C promoter in a cooperative fashion. This study provides the first characterization of this downstream HMG I-C promoter. PMID- 10471824 TI - Proteinase 3 mRNA expression is induced in monocytes but not in neutrophils of patients with cystic fibrosis. AB - Proteinase 3 (PR3), a serine proteinase which can degrade lung tissue, is present in the cystic fibrosis (CF) sputum. In the present study, PR3 protein and mRNA expression was determined in circulating neutrophils and monocytes. CF neutrophils contained similar PR3 concentrations as healthy controls and poorly expressed PR3 mRNA. In contrast, CF monocytes showed significantly higher PR3 concentrations than controls, together with an upregulation of PR3 mRNA expression especially during pulmonary exacerbation. Interestingly, antibiotic treatment fully abrogated PR3 mRNA expression and decreased PR3 protein in monocytes. Our findings highlight a potential role of monocyte-derived PR3 in CF associated airway inflammation. PMID- 10471825 TI - Repression and activation of transcription of phosphoenolpyruvate carboxykinase gene during liver development. AB - Transcriptional activation of the hepatic phosphoenolpyruvate carboxykinase (PEPCK) gene at birth is critical since PEPCK appearance initiates hepatic gluconeogenesis. A delayed appearance results in hypoglycemia, while a premature appearance results in neonatal diabetes, both are incompatible with sustaining life. Experiments using transgenic mice and transfected hepatoma cells suggest that both repression and activation underlie the correct onset of hepatic PEPCK gene transcription. In transgenic mice, transgenes driven by the proximal PEPCK promoter are prematurely expressed in the fetal liver and over-expressed in the neonatal liver, indicating that sequences upstream of the proximal promoter restrain perinatal expression. In Hepa1c1c7 cells, which mimic the fetal liver, the proximal PEPCK promoter (597 bp) exhibited a 3. 5-10-fold higher activity than longer promoters. Repression of the longer promoter (2000 bp) was diminished upon deletion of the sequence spanning positions(-840) to(- 1116) which contains a PPAR/RXR recognition element. The intact 2000 bp PEPCK promoter could be markedly activated by co-transfecting the transcription factor HNF-1 together with C/EBP. It could be repressed by co-transfection with RXRalpha and adding PPARalpha relieved this inhibition. PMID- 10471826 TI - Interaction between the (-170) CRE and the (-150) CCAAT box is necessaryfor efficient activation of the fibronectin gene promoter by cAMP and ATF-2. AB - The fibronectin promoter contains an ATF/cyclic AMP (cAMP) response element (CRE) site two helical turns upstream of a CCAAT site with which it interacts. We investigated the effects of mutating these (-170) CRE and(-150) CCAAT elements on the promoter activity regulated by three different modulators previously known to act through CRE: ATF-2, cAMP and E1a. While the cooperation seems to play no role in E1a action, integrity of the (-150) CCAAT is necessary for ATF-2 and cAMP efficient activation in a cell-specific manner. These results show that the CRE and CCAAT elements function as a 'composite element' and establish a cell specific function for CRE-CCAAT synergy. PMID- 10471827 TI - Spatial gradients of cellular phospho-proteins. AB - If a protein is rapidly phosphorylated and dephosphorylated at separate cellular locations and protein diffusion is slow, then a spatial gradient of the phosphorylated form of the protein may develop within the cell. We have estimated the potential size of such gradients using measured values of protein diffusion coefficients and protein kinase and phosphatase activities. We analysed two different cellular geometries: (1) where the kinases is located on the plasma membrane of a spherical cell and the phospatase is distributed homogenously in the cytoplasm and (2) where the kinase is located on one planar membrane and the phosphatase on a second parallel planar membrane. The estimated gradients of phospho-proteins were potentially very large, which has important implications for cellular signalling. PMID- 10471828 TI - Diadenosine polyphosphates and the control of cyclic AMP concentrations in isolated rat liver cells. AB - Extracellular diadenosine polyphosphates (Ap(n)A), through their interactions with appropriate P(2) receptors, influence a diverse range of intracellular activities. In particular, Ap(4)A stimulates alterations in intracellular calcium homeostasis and subsequent activation of glycogen breakdown in isolated liver cells. Here we show that, like ATP, Ap(4)A and other naturally occurring diadenosine polyphosphates attenuate glucagon-stimulated accumulation of cyclic AMP in isolated rat liver cells. The characteristics of Ap(4)A- and ATP-dependent modulation of glucagon-stimulated cyclic AMP accumulation are similar. These results are discussed in the context of the repertoire of intracellular signalling processes modulated by extracellular nucleotides. PMID- 10471829 TI - Novel cathelicidins in horse leukocytes(1). AB - Cathelicidins are precursors of defense peptides of the innate immunity and are widespread in mammals. Their structure comprises a conserved prepropiece and an antimicrobial domain that is structurally varied both intra- and inter-species. We investigated the complexity of the cathelicidin family in horse by a reverse transcription-PCR-based cloning strategy of myeloid mRNA and by Southern and Western analyses. Three novel cathelicidin sequences were deduced from bone marrow mRNA and designated equine cathelicidins eCATH-1, eCATH-2 and eCATH-3. Putative antimicrobial domains of 26, 27 and 40 residues with no significant sequence homology to other peptides were inferred at the C-terminus of the sequences. Southern analysis of genomic DNA using a probe based on the cathelicidin-conserved propiece revealed a polymorphic DNA region with several hybridization-positive fragments and suggested the presence of additional genes. A null eCATH-1 allele was also demonstrated with a frequency of 0.71 in the horse population analyzed and low amounts of eCATH-1-specific mRNA were found in myeloid cells of gene-positive animals. A Western analysis using antibodies to synthetic eCATH peptides revealed the presence of eCATH-2 and eCATH-3 propeptides, but not of eCATH-1-related polypeptides, in horse neutrophil granules and in the secretions of phorbol myristate acetate-stimulated neutrophils. These results thus suggest that eCATH-2 and eCATH-3 are functional genes, whereas eCATH-1 is unable to encode a polypeptide. PMID- 10471830 TI - Molecular cloning and characterization of AtTERT, a telomerase reverse transcriptase homolog in Arabidopsis thaliana. AB - On the basis of its predicted homology to human telomerase reverse transcriptase (hTERT), a cDNA for Arabidopsis thaliana TERT (AtTERT) has now been isolated from cultured cells. The cDNA contains an open reading frame of 3372 bp, encoding a protein with a predicted size of 131 kDa and isoelectric point of 9.9. The AtTERT protein contains the conserved reverse transcriptase motifs 1, 2 and A-E as well as the TERT-specific T motif. Reverse transcription-polymerase chain reaction analysis and an assay of telomerase activity revealed that both AtTERT mRNA and telomerase activity are abundant in shoot apical meristems but are not detectable in rosette leaves. PMID- 10471831 TI - Distinct roles of profilin in cell morphological changes: microspikes, membrane ruffles, stress fibers, and cytokinesis. AB - Here we report the functional importance of profilin in various actin-mediated morphological changes using H119E mutant profilin I, which is deficient only in actin binding. In the case of actin-protrusive structures from the plasma membrane, H119E-profilin was shown to suppress the formation of Cdc42-induced actin microspikes and Rac-induced membrane ruffles. Conversely, Rho-induced stress fiber formation seemed to occur independently of H119E-profilin introduction. Furthermore, H119E-profilin blocked cleavage furrow ingression and subsequent adhesion to the substratum during cell division, a process in which actin plays indispensable roles. PMID- 10471832 TI - Immunolocalization of cyclooxygenase-2 in the macula densa of human elderly. AB - To gain insight into the role of prostanoids in human kidney function, we examined the distribution of cyclooxygenase (COX) 1 and COX-2 by immunofluorescence and immunohistochemistry in human kidneys from adults of various age groups. COX-1 was detected in the collecting ducts, thin loops of Henle and portions of the renal vasculature. COX-2 was detected in the renal vasculature, medullary interstitial cells, and the macula densa. In addition, COX 2 immunoreactivity was noted in afferent arteries and the macula densa of the renal cortex and was more evident in the kidneys of older adults. PMID- 10471833 TI - A zinc-finger transcription factor induced by TGF-beta promotes apoptotic cell death in epithelial Mv1Lu cells. AB - Transforming growth factor-beta (TGF-beta) superfamily members constitute a group of multifunctional factors that are able to stimulate apoptotic cell death in a variety of cells. In this report, we show that a zinc-finger transcription factor (TIEG) is an immediate early gene transcriptionally induced by TGF-beta in the epithelial Mv1Lu cell line. We also demonstrate that, mimicking TGF-beta effects, ectopic overexpression of TIEG is sufficient to trigger the apoptotic cell program in these cells, which is preceded by a decrease of Bcl-2 protein levels. Finally, apoptotic events elicited by TIEG overexpression can be effectively prevented by ectopic co-expression of Bcl-2. On the basis of these results we suggest that induction of TIEG expression has a role in the pro-apoptotic properties of TGF-beta. PMID- 10471834 TI - Polypeptide release factor eRF1 from Tetrahymena thermophila: cDNA cloning, purification and complex formation with yeast eRF3. AB - The first cDNA for the translational release factor eRF1 of ciliates was cloned from Tetrahymena thermophila. The coding frame contained one UAG and nine UAA codons that are reassigned for glutamine in Tetrahymena. The deduced protein sequence is 57% identical to human eRF1. The recombinant Tetrahymena eRF1 purified from a yeast expression system was able to bind to yeast eRF3 as do other yeast or mammalian eRF1s as a prerequisite step for protein termination. The recombinant Tetrahymena eRF1, nevertheless, failed to catalyze polypeptide termination in vitro with rat or Artemia ribosomes, at least in part, due to less efficient binding to the heterologous ribosomes. Stop codon specificity and phylogenetic significance of Tetrahymena eRF1 are discussed from the conservative protein feature. PMID- 10471835 TI - Dissociation of the eukaryotic initiation factor-4E/4E-BP1 complex involves phosphorylation of 4E-BP1 by an mTOR-associated kinase. AB - mTOR immunoprecipitates contain two 4E-BP1 protein kinase activities. One appears to be due to mTOR itself and results in the phosphorylation of 4E-BP1 on residues T(36) and T(45), as shown previously by others. The other is a kinase which can be separated from mTOR and which phosphorylates 4E-BP1 within a peptide(s) containing residues S(64) and T(69). This phosphorylation, which occurs predominantly on S(64), results in the dissociation of 4E-BP1 from eIF-4E. PMID- 10471836 TI - Microcystin affinity purification of plant protein phosphatases: PP1C, PP5 and a regulatory A-subunit of PP2A. AB - Proteins of approximately 35, 55 and 65kDa were purified from cauliflower extracts by microcystin-Sepharose chromatography and identified by amino acid sequencing as plant forms of protein (serine/threonine) phosphatase 1 (PP1) catalytic subunit, PP5 and a regulatory A-subunit of PP2A, respectively. Peptides that corresponded both to the tetratricopeptide (TPR) repeat and catalytic domains of PP5 were identified. Similar to mammalian PP5,the casein phosphatase activity of plant PP5 was activated >10-fold by arachidonic acid, with half maximal stimulation occurring at approximately 100 microM lipid. PMID- 10471837 TI - Functional studies of a glucagon receptor isolated from frog Rana tigrina rugulosa: implications on the molecular evolution of glucagon receptors in vertebrates. AB - In this report, the first amphibian glucagon receptor (GluR) cDNA was characterized from the liver of the frog Rana tigrina rugulosa. Functional expression of the frog GluR in CHO and COS-7 cells showed a high specificity of the receptor towards human glucagon with an EC(50) value of 0.8+/-0.5 nM. The binding of radioiodinated human glucagon to GluR was displaced in a dose dependent manner only with human glucagon and its antagonist (des-His(1)-[Nle(9) Ala(11)-Ala(16)]) with IC(50) values of 12.0+/-3. 0 and 7.8+/-1.0 nM, respectively. The frog GluR did not display any affinity towards fish and human GLP-1s, and towards glucagon peptides derived from two species of teleost fishes (goldfish, zebrafish). These fish glucagons contain substitutions in several key residues that were previously shown to be critical for the binding of human glucagon to its receptor. By RT-PCR, mRNA transcripts of frog GluR were located in the liver, brain, small intestine and colon. These results demonstrate a conservation of the functional characteristics of the GluRs in frog and mammalian species and provide a framework for a better understanding of the molecular evolution of the GluR and its physiological function in vertebrates. PMID- 10471838 TI - Diverse PAH transcripts in lymphocytes of PKU patients with putative nonsense (G272X, Y356X) and missense (P281L, R408Q) mutations. AB - The majority of mutations in the human phenylalanine hydroxylase (PAH) gene that lead to the recessive disease phenylketonuria (PKU) are believed to affect the activity or stability of the PAH enzyme. In this study we have performed in vivo analyses of lymphocyte PAH mRNA from PKU patients homozygous for the PKU missense mutations P281L and R408Q as well as the nonsense mutations G272X and Y356X. The mutations G272X, P281L and R408Q, which are located outside the consensus splice site sequence, result in transcripts with one or more exons skipped in addition to full-length transcripts. The mutation Y356X results in transcripts with one or more exons skipped, but no full-length transcripts. Our findings question the value of functional and structural predictions of mutations at the protein level without analyses of the corresponding transcript. PMID- 10471839 TI - Molecular cloning and sequencing of two 'short chain' and two 'long chain' K(+) channel-blocking peptides from the Chinese scorpion Buthus martensii Karsch. AB - Five full-length cDNAs encoding the precursors of two 'short chain' scorpion non toxic peptides active on Ca(2+)-activated K(+) channels (BmP02 and BmP03) and two novel putative long chain K(+) channel-blocking peptides (named BmTXKbeta and BmTXKbeta2) were first isolated from the venom gland cDNA library of the Chinese scorpion Buthus martensii Karsch (BmK). BmTXKbeta2 showed a high similarity with AaTXKbeta, while BmTXKbeta was completely different in the deduced primary structure from the long chain and short chain scorpion toxins already characterized. Thus, BmTXKbeta expands the scorpion long chain K(+) channel blocking peptide family. Although little sequence similarity exists between the above two short and two long peptides, they are similar at the positions of six cysteines, suggesting that they should all share a similar scaffold composed of an alpha-helix and a three-stranded beta-sheet. PMID- 10471840 TI - Dynamic re-distribution of protein kinase D (PKD) as revealed by a GFP-PKD fusion protein: dissociation from PKD activation. AB - Protein kinase D (PKD)/protein kinase Cmicro (PKCmicro, a serine/threonine protein kinase with distinct structural and enzymological properties, is rapidly activated in intact cells via PKC. The amino-terminal region of PKD contains a cysteine-rich domain (CRD) that directly binds phorbol esters with a high affinity. Here, we show that treatment of transfected RBL 2H3 cells with phorbol 12,13-dibutyrate (PDB) induces a striking CRD-dependent translocation of PKD from the cytosol to the plasma membrane, as shown by real time visualization of a functional green fluorescent protein (GFP)-PKD fusion protein. A single amino acid substitution in the second cysteine-rich motif of PKD (P287G) prevented PDB induced membrane translocation but did not affect PKD activation. Our results indicate that PKD translocation and activation are distinct processes that operate in parallel to regulate the activity and localization of this enzyme in intact cells. PMID- 10471841 TI - Identification of melanin concentrating hormone (MCH) as the natural ligand for the orphan somatostatin-like receptor 1 (SLC-1). AB - To identify possible ligands of the orphan somatostatin-like receptor 1 (SLC-1), rat brain extracts were analyzed by using the functional expression system of Xenopus oocytes injected with cRNAs encoding SLC-1 and G protein-gated inwardly rectifying potassium channels (GIRK). A strong inward current was observed with crude rat brain extracts which upon further purification by cation exchange chromatography and high performance liquid chromatography (HPLC) yielded two peptides with a high agonist activity. Mass spectrometry and partial peptide sequencing revealed that one peptide is identical with the neuropeptide melanin concentrating hormone (MCH), the other represents a truncated version of MCH lacking the three N-terminal amino acid residues. Xenopus oocytes expressing the MCH receptor responded to nM concentrations of synthetic MCH not only by the activation of GIRK-mediated currents but also by the induction of Ca(2+) dependent chloride currents mediated by phospholipase C. This indicates that the MCH receptor can couple either to the G(i)- or G(q)-mediated signal transduction pathway, suggesting that MCH may serve for a number of distinct brain functions including food uptake behavior. PMID- 10471844 TI - Auditory priming within and across modalities: evidence from positron emission tomography. AB - Previous neuroimaging studies of perceptual priming have reported priming-related decreases in the extrastriate cortex. However, because these experiments have used visual stimuli, it is unclear whether the observed decreases are associated specifically with some aspect of visual perceptual processing or with more general aspects of priming. We studied within- and cross-modality priming using an auditory word stem completion paradigm. Positron emission tomography (PET) images were obtained during stem completion and a fixation task. Within-modality auditory priming was associated with blood flow decreases in the extrastriate cortex (bilateral), medial/right anterior prefrontal cortex, right angular gyrus, and precuneus. In cross-modality priming, the study list was presented visually, and subjects completed auditory word stems. Cross-modality priming was associated with trends for blood flow decreases in the left angular gyrus and increases in the medial/right anterior prefrontal cortex. Results thus indicate that reduced activity in the extrastriate cortex accompanies within-modality priming in both visual and auditory modalities. PMID- 10471845 TI - Can face recognition really be dissociated from object recognition? AB - We argue that the current literature on prosopagnosia fails to demonstrate unequivocal evidence for a disproportionate impairment for faces as compared to nonface objects. Two prosopagnosic subjects were tested for the discrimination of objects from several categories (face as well as nonface) at different levels of categorization (basic, subordinate, and exemplar levels). Several dependent measures were obtained including accuracy, signal detection measures, and response times. The results from Experiments 1 to 4 demonstrate that, in simultaneous-matching tasks, response times may reveal impairments with nonface objects in subjects whose error rates only indicate a face deficit. The results from Experiments 5 and 6 show that, given limited stimulus presentation times for face and nonface objects, the same subjects may demonstrate a deficit for both stimulus categories in sensitivity. In Experiments 7, 8 and 9, a match-to-sample task that places greater demands on memory led to comparable recognition sensitivity with both face and nonface objects. Regardless of object category, the prosopagnosic subjects were more affected by manipulations of the level of categorization than normal controls. This result raises questions regarding neuropsychological evidence for the modularity of face recognition, as well as its theoretical and methodological foundations. PMID- 10471846 TI - Delineating necessary and sufficient neural systems with functional imaging studies of neuropsychological patients. AB - This paper demonstrates how functional imaging studies of neuropsychological patients can provide a way of determining which areas in a cognitive network are jointly necessary and sufficient. The approach is illustrated with an investigation of the neural system underlying semantic similarity judgments. Functional neuroimaging demonstrates that normal subjects activate left temporal, parietal, and inferior frontal cortices during this task relative to physical size judgments. Neuropsychology demonstrates that damage to the temporal and parietal regions results in semantic deficits, indicating that these areas are necessary for task performance. In contrast, damage to the inferior frontal cortex does not impair task performance, indicating that the inferior frontal cortex might not be necessary. However, there are two other possible accounts of intact performance following frontal lobe damage: (1) there is functional reorganization involving the right frontal cortex and (2) there is peri-infarct activity around the damaged left-hemisphere tissue. Functional imaging of the patient is required to discount these possibilities. We investigated a patient (SW), who was able to associate words and pictures on the basis of semantic relationships despite extensive damage to the left frontal, inferior parietal, and superior temporal cortices. Although SW showed peri-infarct activation in left extrasylvian temporal cortices, no activity was observed in either left or right inferior frontal cortices. These findings demonstrate that activity in extrasylvian temporo-parietal and medial superior frontal regions is sufficient to perform semantic similarity judgments. In contrast, the left inferior frontal activations detected in each control subject appear not to be necessary for task performance. In conclusion, necessary and sufficient brain systems can be delineated by functional imaging or brain-damaged patients who are not functionally impaired. PMID- 10471847 TI - The neural circuitry involved in the reading of German words and pseudowords: A PET study. AB - Silent reading and reading aloud of German words and pseudowords were used in a PET study using (15O)butanol to examine the neural correlates of reading and of the phonological conversion of legal letter strings, with or without meaning. The results of 11 healthy, right-handed volunteers in the age range of 25 to 30 years showed activation of the lingual gyri during silent reading in comparison with viewing a fixation cross. Comparisons between the reading of words and pseudowords suggest the involvement of the middle temporal gyri in retrieving both the phonological and semantic code for words. The reading of pseudowords activates the left inferior frontal gyrus, including the ventral part of Broca's area, to a larger extent than the reading of words. This suggests that this area might be involved in the sublexical conversion of orthographic input strings into phonological output codes. (Pre)motor areas were found to be activated during both silent reading and reading aloud. On the basis of the obtained activation patterns, it is hypothesized that the articulation of high-frequency syllables requires the retrieval of their concomitant articulatory gestures from the SMA and that the articulation of low-frequency syllables recruits the left medial premotor cortex. PMID- 10471848 TI - The neural substrate of picture naming. AB - A PET study of 10 normal males was carried out using the bolus H215O intravenous injection technique to examine the effects of picture naming and semantic judgment on blood flow. In a series of conditions, subjects (1) passively viewed flashing plus signs, (2) noted the occurrence of abstract patterns, (3) named animal pictures, or (4) carried out a semantic judgment on animal pictures. Anticipatory scans were carried out after the subjects were presented with the instructions but before they began the cognitive task, as they were passively viewing plus signs. Our results serve to clarify a number of current controversies regarding the neural substrate of picture naming. The results indicate that the fusiform gyrus is unlikely to be the region where low-level perceptual processing such as shape analysis is undertaken. In fact, our evidence suggests that activation of the fusiform gyrus is most likely related to visual perceptual semantic processing. In addition, the inferior/middle frontal lobe activity observed while performing the picture naming and semantic judgment tasks does not appear to be due to the effects of anticipation or preparation. Furthermore, there appears to be a set of regions (a semantic network) that becomes activated regardless of whether the subjects perform a picture naming or semantic judgment task. Finally, picture naming of animals did not activate either parietal regions or anterior inferior left temporal regions, regardless of what subtraction baseline was used. PMID- 10471849 TI - Interactive processing of sensory input and motor output in the human hippocampus. AB - Recent studies of visuomotor integration suggest that the motor system may be intimately involved in the detection of salient features of the sensory scene. The final stages of sensory processing occur in hippocampal structures. We measured human neuromagnetic responses during motor reaction to an auditory cue embedded in high-speed multimodal stimulation. Our results demonstrate that large scale cognitive networks may recruit additional resources from the hippocampus during sensorimotor integration. Hippocampal activity from 300 msec before to 200 msec after cued movements was enhanced significantly over that observed during self-paced movements. The dominant hippocampal activity appeared equally synchronized to both sensory input and motor output, consistent with timing by an intrinsic mechanism, possibly provided by ongoing theta oscillations PMID- 10471850 TI - Short- and long-delay intracranial ERP repetition effects dissociate memory systems in the human brain. AB - Prior exposure to a stimulus can facilitate the performance to subsequent presentations of that stimulus. ERP studies have shown that this facilitation is associated with the modulation of two components (N400 and P600). Investigation of the time course of both behavioral and ERP repetition effects have led to the assumption that it is subserved by the combination of at least two mechanisms operating at different time-points: a short-delay semantic activation and a long lasting episodic mechanism. The present experiment recorded intracranial ERPs from various structures during a continuous recognition memory task to investigate the respective contribution of the different brain regions to short- and long-delay ERP repetition effects. The results are in good agreement with both the classical neuropsychological literature and the more recent data obtained with functional imagery techniques. They provide electrophysiological evidence of multiple anatomo-functional memory systems in the human brain: a short-term semantic activation system and a long-term episodic memory system, with interface structures that coordinate the functioning of these two systems. PMID- 10471851 TI - Illusory contour perception and amodal boundary completion: evidence of a dissociation following callosotomy. AB - A fundamental problem in form perception is how the visual system can link together spatially separated contour fragments to form the percept of a unitary shape. Illusory contours and amodal completion are two phenomena that demonstrate this linking process. In the present study we investigate these phenomena in the divided hemispheres of two callosotomy ("split-brain") patients. The data suggest that dissociable neural mechanisms are responsible for the generation of illusory contours and amodal completion. Although both cerebral hemispheres appear to be equally capable of perceiving illusory contours, amodal completion is more readily utilized by the right hemisphere. These results suggest that illusory contours may be attributable to low-level visual processes common to both hemispheres, whereas amodal completion reflects a higher-level, lateralized process. PMID- 10471852 TI - Multichannel cochlear nucleus stimulation. AB - Stimulation of the cochlear nucleus (CN) has been used on a limited basis for rehabilitation of a select group of patients with bilateral acoustic neuromas. These patients were implanted with an electrode placed on the surface of the CN after resection of their tumors. Animal studies have demonstrated greater efficiency of a penetrating CN electrode in activating the central auditory system than a surface electrode. The objective of this work was to study the electrically evoked middle latency response generated by stimulation through a penetrating multichannel CN electrode in an animal model. Six pigmented guinea pigs underwent implantation with a penetrating multichannel CN electrode. Threshold, latency, and input-output functions of electrically evoked middle latency responses with different stimulation pads were studied. There were systematic differences in the latency and amplitude of the input-output functions depending on the site of stimulation within the CN. The results support the hypothesis that discrete activation of neuronal subpopulations within the CN is possible with a penetrating multichannel microelectrode. PMID- 10471853 TI - Standardized format for depicting hearing preservation results in the management of acoustic neuroma. AB - The Committee on Hearing and Equilibrium of the American Academy of Otolaryngology-Head and Neck Surgery recently published guidelines for reporting hearing preservation in the treatment of acoustic neuromas. These suggestions included pretreatment and posttreatment pure-tone hearing thresholds, word recognition scores, and hearing classification. We present a standardized reporting format that addresses the Committee's recommendations and displays individual patient audiologic data as a simple, concise plot of posttreatment hearing results. To illustrate the use of the recommended format, preoperative and postoperative hearing data from our institution are reported. Such reporting criteria will facilitate comparative reviews of studies of hearing preservation after surgical or radiotherapeutic management of acoustic neuromas, while providing specific data for individual patient outcome analysis. PMID- 10471854 TI - Role of motor unit number estimate electromyography in experimental canine laryngeal reinnervation. AB - Laryngeal electromyography has been used clinically to differentiate neuromuscular pathology from other causes of vocal fold immobility such as arytenoid dislocation, tumor invasion, or cricoarytenoid joint fixation. Electromyography has also been used to predict the prognosis for nerve recovery in laryngeal paralysis. Existing electromyographic techniques either record activity with voluntary motion or study nerve conduction. In this study a new technique, motor unit number estimation, a commercially available quantitative method of electromyographic analysis, is used to study the progress of recovery of vocal fold function after recurrent laryngeal nerve injury. Four dogs underwent transection and immediate reanastomosis of selected branches of the adductor and abductor branches of the recurrent laryngeal nerve on 1 side; the opposite side served as a control. Baseline electromyographic and videolaryngoscopic studies were performed. These measures were then repeated in a longitudinal fashion every 6 weeks after denervation. The motor unit number estimation technique indicated a return of motor unit numbers with time, along with estimates of their size. This was consistent with the expected progress of laryngeal reinnervation. These data and their predictive value for nerve recovery will be discussed. PMID- 10471855 TI - Thermal myringotomy for eustachian tube dysfunction in hyperbaric oxygen therapy. AB - Otolaryngologists are frequently asked to manage eustachian tube dysfunction (ETD) in patients undergoing hyperbaric oxygen therapy (HBO). HBO patients with intractable ETD currently are treated by tympanostomy tube placement; typically, these tubes are indwelling far longer than is required by the duration of HBO. Also, tubes in this population are associated with higher complication rates of persistent perforation and otorrhea. We investigated the use of thermal myringotomy as an alternative to tympanostomy tube placement in this clinical setting. Potentially, thermal myringotomy avoids the risks and complications associated with indwelling tympanostomy tubes and would be a temporally more appropriate treatment during short- and intermediate-term HBO. In this study 13 patients undergoing HBO who would have required tympanostomy tube placement instead underwent bilateral thermal myringotomies. At the fifth postoperative week, 96% of myringotomies were patent; this duration is adequate for most HBO courses. No patient required a second myringotomy for premature closure. The persistent perforation rate was 15% (at the end of 6 months), which compares favorably with the rate observed with tympanostomy tubes in this unique population of poor wound healers. Only 1 patient had otorrhea; this resolved with dry ear precautions. This study demonstrates thermal myringotomy to be an effective technique for middle ear ventilation in patients undergoing HBO in whom ETD develops. PMID- 10471856 TI - Impact of resistant pneumococcus on rates of acute mastoiditis. AB - OBJECTIVE: To determine whether the rate of acute mastoiditis is rising, specifically as a result of antibiotic-resistant strains of Streptococcus pneumoniae. METHODS: A retrospective chart review of all patients with a discharge diagnosis of acute mastoiditis between July 1, 1987, and June 30, 1997, was performed at our academic, tertiary-care medical center. There were no interventions, and the main outcome measures included the number of cases of acute or coalescent mastoiditis, stratified by pathogen, per year. RESULTS: The rate of acute mastoiditis as a proportion of yearly otorhinolaryngology admissions increased linearly over time (P = 0. 024). Pneumococcal-related rates of acute mastoiditis, expressed as a proportion of yearly hospital and otorhinolaryngology admissions, increased linearly over time (P = 0.002, P = 0.002). All but 1 case of pneumococcal mastoiditis during the past 3 years were caused by penicillin-resistant strains. CONCLUSIONS: The emergence of antibiotic resistant S pneumoniae may be responsible for an increasing rate of acute mastoiditis. PMID- 10471857 TI - Clinical and histologic evaluation of an indwelling, inflatable, long-term laryngeal stent in the canine model. AB - OBJECTIVE: The goal was assessment of a revolutionary approach to laryngeal stenting. A radically designed, atraumatic, inflatable, low-pressure, high volume, endoscopically inserted laryngeal stent for effective intralaryngeal support or long-term prevention of aspiration was prospectively evaluated in a canine model to reveal any significant short- and long-term anatomic and functional changes in the larynx. METHODS: A self-sustaining, tube-free, long term flap tracheostomy was performed in 14 dogs, followed by endoscopic laryngeal stent insertion. The stents were inflated, and their intraluminal pressures were monitored daily. The stenting period ranged between 3 and 46 days (mean 27.4 days). The animals were then euthanized, and total laryngectomy was performed. The larynges underwent gross and microscopic examinations, and a grading scale of reactive changes caused by the stent placement was created on the basis of the findings. RESULTS: Dye studies for aspiration were negative in all animals. Gross findings consisted of varying degrees of small ulceration, localized polypoid lesions, and granulomas. Grossly, 9 dogs exhibited small superficial ulceration of the true vocal cords and posterior commissure. Six dogs developed minor polyps or polypoid changes and/or granulomas, respectively. No gross changes were seen in 2 of the dogs. Histopathologically, 10 dogs demonstrated grade I or II histopathologic changes. Only 4 of the dogs exhibited inflammation extending into the underlying cartilages, grade III. One dog did not complete the observation period. CONCLUSION: This new stent demonstrated safety and biocompatibility with minimal local tissue reaction to its extended long-term placement. Good tolerance to the stent was documented, with minimal side effects similar to those that would be observed in human subjects after endotracheal intubation. When followed up in human patients, such minimal lesions have not resulted in significant long lasting functional impairments. These initial results would indicate that this stent is well tolerated for long-term application and qualify as the preferred device for both management of aspiration and postoperative endolaryngeal bolstering support. PMID- 10471858 TI - Benefits of routine maxillectomy and orbital reconstruction with the rectus abdominis free flap. AB - Maxillary defects lead to functional and cosmetic deficiencies, especially with tumor invasion of the orbit and cribriform plate. Additionally problematic is the near-total palatal resection in patients with poor dentition (ie, not useful for anchoring obturators) and the desire for return of deglutition and useful speech. A series of 12 patients is presented in whom a rectus abdominis free flap was used for reconstruction of the palate, maxilla, and/or orbit. Nine of 12 free flaps were used to reconstruct defects involving the maxilla and orbit; all were successful in restoring hard palate continuity and function without the use of an obturator. The remaining 3 patients underwent successful repair of defects involving the orbit and medial maxilla without complication. Four of those 9 patients with palatal reconstruction augmented their dental rehabilitation with either their existing upper denture plate or the use of a cosmetic appliance anchored to their remaining dentition. Objectively, the rectus abdominis myocutaneous free flap proved to be a reliable and expeditious method of restoring function and acceptable cosmesis. We believe this technique should be routinely considered in the reconstruction of these defects. PMID- 10471859 TI - Facial nerve function after partial superficial parotidectomy: An 11-year review (1987-1997). AB - Permanent facial nerve dysfunction is a potential complication of every parotid surgery. Partial superficial parotidectomy, a conservative resectioning that requires neither dissection of the full facial nerve nor excision of the superficial lobe, produces lower rates of facial nerve dysfunction and soft tissue deformity than the traditional method. This report describes a single surgeon's experience with partial superficial parotidectomy from 1987 to 1997. Fifty-nine patients with mobile, benign, and low-grade malignant tumors, limited to the superficial lobe, underwent partial superficial parotidectomy with selective nerve dissection. Adequate margins were obtained, based on the premise that the tumor-to-nerve margin is often the true one. No patients had permanent nerve paralysis or paresis, and only 10 incurred transient facial nerve paresis. Age, histology, and sex were not significant factors in postoperative facial nerve function. No patients had recurrences. PMID- 10471860 TI - Training the physician-scholar in otolaryngology-head and neck surgery. AB - The Resident Education Committee of the Society of University Otolaryngologists constructed a questionnaire for young academic otolaryngologist-head and neck surgeons to better understand their training background and to garner their opinions concerning adequacy or deficiencies in various aspects of that training. The questionnaire was mailed to 145 individuals who were in academic posts for 5 years or fewer in 1997. There was an overall response rate of 88.3%. Of the 128 respondents, 89% identified additional training, most commonly a clinical fellowship, in preparation for an academic career. The median number of hours per week devoted to professionally related activity was 61, of which two thirds was spent in direct patient care. The most common source of funds to pursue research activities was intradepartmental resources. Most individuals were satisfied with their jobs, although one quarter were considering leaving academic practice within the year. The single most important reason motivating selection of an academic career was a desire to teach. Details of the specific training and competencies and recommendations for improvement in resident training were obtained. Specific recommendations were generated for improving the training of future academic otolaryngologists; these recommendations include clarity of job description, a single track for clinical training for academicians and nonacademicians, more training in pertinent skills including research training, protected time for research, and amelioration of some of the downsides of academic life. PMID- 10471861 TI - Effects of vasoactive topical agents on the survival of dorsal skin flaps in rats. AB - OBJECTIVE: To evaluate 3 vasoactive topical agents regarding their ability to improve the survival of random skin flaps in rats. METHOD: Thirty-five adult Sprague-Dawley rats had a single 9 x 2 cm random skin flap raised with topical application of a white petroleum ointment (control group), prostaglandin E2, minoxidil, or nitroglycerin to the flap. The effect of the agents on the viability of the flaps was evaluated. RESULTS: The control group had an average survival area of 60.8% +/- 7.9%. The nitroglycerin and minoxidil groups were not significantly different, with an average area of survival of 60.1% +/- 8.1% and 63.3% +/- 4.7%, respectively. The prostaglandin E2 group had average area of survival of 74.2% +/- 10.5% (P = 0.002). CONCLUSION: Prostaglandin E2 demonstrated a significant increase in flap survival. Nitroglycerin ointment and minoxidil had no beneficial effects when compared with control. PMID- 10471862 TI - Evidence of possible localized specific immunoglobulin E production in middle ear fluid as demonstrated by ELISA testing. AB - To determine whether IgE in the middle ear represents a passive transudate from the serum or may be produced within the middle ear itself, paired effusion and serum from 18 atopic children with otitis media with effusion were tested by micro-ELISA for specific IgE to 12 allergens. Elevated effusion concentrations of specific IgE were present in 83.3% (15 of 18) of atopic patients, but only 30% (64 of 214) of the serum antibodies appeared in the effusion. The data show that in atopic children with otitis media with effusion, there is no relation between a patient's serum and effusion level of IgE for specific antibodies (P < 0.001), suggesting that IgE in middle ear effusion is not a transudate but more likely reflects an active localized process in atopic patients. PMID- 10471863 TI - Effects of dimethylthiourea, melatonin, and hyperbaric oxygen therapy on the survival of reimplanted rabbit auricular composite grafts. AB - Microvascular reconstruction currently offers the best chance of survival for an ear segment reimplanted after amputation. Unfortunately, this technique is possible only when ear and scalp vessels remain intact. Direct reattachment of the amputated segment as a composite graft is less reliable because survival is poor for grafts larger than 2 cm in diameter. However, if survival could be improved, direct reattachment would be an attractive alternative in situations in which microsurgical revascularization is not possible. Certain pharmacologic agents have been shown to enhance the survival of composite grafts. This study demonstrated that hyperbaric oxygen, dimethylthiourea, and melatonin significantly affected the survival of reimplanted auricular composite grafts at day 7. However, by day 21 the average percentage of survival for all groups approached 13% to 14%. Dimethylthiourea had the most beneficial effect on survival early in the postoperative period, whereas the hyperbaric oxygen group demonstrated the poorest survival. PMID- 10471864 TI - Interactive effects of the middle ear pathology and the associated hearing loss on transient-evoked otoacoustic emission measures. AB - Sixty-three children aged 4 to 17 years were examined by tympanometry, pure-tone audiometry, transient-evoked otoacoustic emissions (TEOAEs), and otoscopy to evaluate the effects of middle ear pathologies and the associated hearing loss on TEOAEs. TEOAE measures were highly specific (93.8%) in identifying normal ears that passed both audiometric and tympanometric criteria. The sensitivity for identifying abnormal ears was also reasonable (83. 3%). The effects of the middle ear abnormality were most significant, regardless of the degree of hearing loss, when the tympanogram was type B with normal volume measures, which is associated with reduced eardrum mobility and middle ear fluid. The middle ear conditions producing the greater negative pressure, which in turn led to more conductive hearing loss, also produced more TEOAE failures. The mere presence of an open ventilation tube was not a determining factor for absent TEOAEs because 60% of the open ventilation tubes had normal TEOAEs. Provided that the clinician understands the effects of middle ear pathologies on otoacoustic emissions, TEOAEs can be a great asset for diagnosis of both otologic and audiologic disorders. PMID- 10471865 TI - Transplanted fetal fibroblasts: survival and distribution over time in normal adult dermis compared with autogenic, allogenic, and xenogenic adult fibroblasts. AB - Cell therapy is a widely applicable therapeutic approach using cells and cell elements, frequently from fetal or young animals, for their beneficial effects. This study evaluated the host response to and tolerance of transplanted fetal skin fibroblasts. Cultured fibroblasts from adult rabbit skin (autogenic and allogenic), 21-day fetal rabbit skin (allogenic), and adult pig skin (xenogenic) were labeled with a fluorescent vital dye CM-DiI, injected intradermally into the dorsal skin of adult rabbits at multiple sites and then biopsied over an 8-week period. Each cell type showed a biphasic distribution curve with an early phase (0 to 28 days) and a late phase (28 to 56 days). In the early phase, cells showed a rise and fall in total cell density (reflecting an increase and then a decrease in total cell number), followed by a slow decrease in cell density with cells still detectable at 56 days. Fetal cells showed the highest survival at the end of the study. None of the groups showed clinical or histologic signs of acute inflammation or rejection. This study demonstrated that (1) transplanted fibroblasts are well tolerated by an immunologically competent host, (2) CM-DiI labeled cells are detectable in vivo for at least 8 weeks, and (3) fetal fibroblasts have a distribution and survival profile that is distinct from that of adult fibroblasts. PMID- 10471866 TI - Mold testing by RAST and skin test methods in patients with allergic fungal sinusitis. AB - RAST tests have traditionally been considered less sensitive than skin tests during investigation of atopy involving molds. This has been attributed to technical problems such as difficulty in binding the mold antigen to the carrier substrate. Ten patients with proven allergic fungal sinusitis were evaluated for sensitivity to 11 important molds by both RAST and dilutional intradermal testing. A predictable correlation between RAST and skin test scores was observed in many, but not all, cases. Most often this disparity was in the form of greater sensitivity indicated by skin testing than by RAST, sometimes differing by as many as 3 classes. The lack of concordance was not confined to testing for the fungi cultured from the sinuses, nor was it more or less pronounced in the case of dematiaceous fungi. The most likely causes for the disparity noted in this series are subtle differences in antigens used in skin test material and for RAST standards. Skin tests allow for evaluation of delayed and late-phase reactions, a measurement not possible by specific IgE testing with RAST. Delayed skin test reactions were not noted in this series of patients. An additional important finding was the sensitivity of patients with allergic fungal sinusitis to virtually every fungal antigen to which they were tested. PMID- 10471867 TI - Outcome of reflux therapy on pediatric chronic sinusitis. AB - OBJECTIVE: The cause of pediatric chronic sinusitis is multifactorial, but nasal edema appears to be the initial pathologic step. The objective of this study is to evaluate gastronasal reflux as a possible cause of pediatric sinusitis. METHODS: Thirty children with chronic sinusitis were believed to be appropriate candidates for functional endoscopic sinus surgery. Children were evaluated retrospectively for their response to reflux therapy with regard to their sinus symptoms and avoidance of sinus surgery. RESULTS: Two of the 30 children were eventually excluded because they were taken to surgery for the specific purpose of contact point release. Chart review at 24-month follow-up indicated that 25 of the 28 children (89%) avoided sinus surgery. CONCLUSION: After reflux treatment, the number of children requiring sinus surgery was dramatically reduced. The results of this preliminary pediatric study indicate that gastronasal reflux should be evaluated and treated before sinus surgical intervention. PMID- 10471868 TI - Predicting airway risk in angioedema: staging system based on presentation. AB - Angioedema is an immunologically mediated, anatomically limited, nonpitting edema that can lead to life-threatening airway obstruction. To predict the risk of airway compromise in angioedema, we retrospectively reviewed 93 episodes in 80 patients from 1985 to 1995. Intubation or tracheotomy was necessary in 9 (9.7%) cases. Angiotensin-converting enzyme inhibitor use in 36 cases (39%) was associated with intensive care unit (ICU) admission (P = 0.05). ICU stay correlated significantly with presentation with voice change, hoarseness, dyspnea, and rash (P < 0.05). Voice change, hoarseness, dyspnea, and stridor were present in patients requiring airway intervention (P < 0.05). On the basis of our data, we propose a staging system by which airway risk may be predicted from the anatomic site of presentation. Patients with facial rash, facial edema, lip edema (stage I), and soft palate edema (stage II) were treated as outpatients and on the hospital ward. Patients with lingual edema (stage III) usually required ICU admission. All patients with laryngeal edema (stage IV) were admitted to the ICU. Airway intervention was necessary in 7% of stage III patients and in 24% of stage IV cases. No deaths were caused by angioedema. Airway risk in angioedema may be predicted by anatomic site of presentation, allowing appropriate triage with preparation for airway intervention in selected cases. PMID- 10471869 TI - Fate of rigid fixation in pediatric craniofacial surgery. AB - The advantages of rigid fixation in adult craniofacial surgery are well documented, and implanted hardware is not routinely removed unless specifically indicated. There is a tendency, however, to remove hardware in children because of concerns with growth restriction, plate migration, and the lack of information on the fate of miniplates when used in pediatric craniofacial surgery. It has been our practice during the past decade not to remove hardware in children unless specifically indicated. Our study included a total of 121 procedures in 96 children, with an average age of 3.9 years and an average follow-up of 5 years. We placed 375 titanium plates and 1944 screws from 3 manufacturers. Complications encountered in children with titanium plates were as follows: 5 cases of delayed growth and 1 instance of restricted growth, 4 screw migrations (none intracranial), 9 palpable plates causing pain, 3 fluid accumulations over plates, 2 cases of meningitis, and 8 instances of plate and screw removal from the above complications. Twenty-two of 96 patients (23%) had a total of 27 complications from 121 procedures (22%). There were 6 cases in which pain precipitated removal of hardware, 1 case of an excessively mobile plate, and 1 case of documented growth restriction requiring removal; therefore our overall reoperation rate for plate removal was 8%, with no intracranial plate or screw migration. PMID- 10471870 TI - Comparison of postoperative cardiopulmonary examinations and chest radiographs to detect pulmonary complications after adult tracheotomy. AB - OBJECTIVE: To prospectively evaluate the effectiveness of clinical cardiopulmonary examination compared with that of chest radiography in detecting postoperative pulmonary complications after tracheotomy in adults. METHODS: One hundred consecutive patients undergoing tracheotomy were evaluated. Pulmonary complications were recorded, and the effectiveness of the physical examination was compared with that of the chest x-ray in detecting these complications. Patient age, sex, diagnosis, urgency of the procedure, and anesthetic technique were evaluated to determine any relationship to postoperative complications. RESULTS: Of the 100 patients, 87 patients underwent postoperative chest radiography, and all patients had postoperative cardiopulmonary examinations. One patient (1%) had a bilateral pneumothorax, which was detected clinically and confirmed by a chest radiograph. Five patients had postoperative pulmonary edema confirmed by clinical examination and radiography. Additional minor complications were noted, including minor bleeding and stomal infection, for an overall complication rate of 13%. CONCLUSIONS: All postoperative pulmonary complications were detected by physical examination. Therefore routine postoperative chest radiographs in uncomplicated tracheotomies are not necessary if a thorough postoperative cardiopulmonary examination is performed. Cost analysis reveals a savings of approximately $19,000 with the proposed criteria for postoperative chest x-ray. PMID- 10471871 TI - Effect of topical antibiotic therapy on recovery after tonsillectomy in adults. AB - OBJECTIVE: Systemic antibiotics given during the first week after tonsillectomy appear to be effective in reducing postoperative morbidity. We assessed the effectiveness of perioperative topical antibiotic rinses in reducing posttonsillectomy morbidity. METHODS: A randomized, double-blinded, placebo controlled pilot study of 36 patients undergoing tonsillectomy was used to evaluate the effects of a standard 7-day systemic regimen of perioperative intravenous ampicillin/oral amoxicillin and 2 single-day topical antibiotic regimens: (1) clindamycin (Cleocin) and (2) amoxicillin/clavulanate (Augmentin) and ticarcillin/clavulanate (Timentin). RESULTS: Mean aerobic and anaerobic oral bacterial counts were decreased in both topical treatment groups compared with the placebo group on the first postoperative day, achieving statistical significance with Augmentin/Timentin (aerobic and anaerobic bacterial counts) and Cleocin (aerobic counts). Significantly less postoperative pain and mouth odor were reported for both Cleocin (P = 0.014 and P = 0.005, respectively) and Augmentin/Timentin (P = 0.026 and P = 0.05, respectively) topical treatment groups when compared with the placebo group. CONCLUSIONS: Preliminary results indicate a reduction in oral bacterial counts and postoperative morbidity in adult patients receiving topical antibiotics compared with patients receiving placebo; further investigation is warranted. PMID- 10471872 TI - Electrocochleographic changes after intranasal allergen challenge: A possible diagnostic tool in patients with Meniere's disease. AB - Numerous observers have suggested a relationship between allergy and Meniere's disease, but objective proof has heretofore been limited. Using standard criteria, we studied a group of 7 patients with previously diagnosed Meniere's disease in whom significant allergy to 1 or more inhalants had also been diagnosed. Patients underwent a baseline electrocochleographic study followed by intranasal challenge with a carefully quantified amount of the allergen to which they were most sensitive. This was followed by a second electrocochleogram. Four of the 7 patients demonstrated at least a 15% increase in the summating potential/action potential ratio in 1 ear, associated with the production of subjective inner ear symptoms. We present this protocol as a potentially useful tool to further study whether inhalant allergy may be a causative factor in patients with Meniere's disease. PMID- 10471873 TI - Unusual otolaryngic presentations of ameloblastoma. PMID- 10471874 TI - Stapes footplate fistula and recurrent meningitis. PMID- 10471875 TI - Black thyroid: A collaborative series. PMID- 10471876 TI - Extramedullary plasmacytoma in the anterior cervical region. PMID- 10471877 TI - External auditory canal cholesteatoma with erosion into the mastoid. PMID- 10471878 TI - Neurosarcoidosis: An unusual case presenting as a cerebellopontine angle tumor. PMID- 10471879 TI - Airway compromise caused by disk battery ingestion. PMID- 10471880 TI - Dysgeusia associated with tonsillectomy. PMID- 10471881 TI - Lingual metastasis as the initial presentation of a large cell lung carcinoma. PMID- 10471882 TI - Assessment of nasal mucosa blood supply by quantitative endoscopic imaging of the back-scattered light. AB - In this article we present a method for the objective assessment and monitoring of tissue blood supply using a specially developed endoscopic imaging colorimeter that enables quantitative color modeling of the back-scattered light during endoscopic examination. Tissue blood volume changes in the nasal mucosa, induced by xylometazoline hydrochloride nasal spray, were evaluated with this method. It was found that quantitative imaging provides sensitive, reproducible, and reliable means for the monitoring and mapping of tissue blood supply and is easy to use routinely. The results showed that saturation decreases with time, being the most sensitive color parameter to the vasoconstriction procedure. It appears that objective indexes for optical tissue characterization and analysis may be promising in the understanding of the pathophysiology of tissue changes and in the objective evaluation of their response to different therapeutic schemes. PMID- 10471883 TI - Does choice of hearing selection criterion and reporting criteria affect the hearing preservation rate in vestibular schwannoma surgery? AB - With increasing refinement in the surgery of vestibular schwannoma the aims of complete tumor removal and facial nerve preservation have been largely fulfilled. However, the reputation of and place for hearing-preservation surgery still remain uncertain. A major part of this uncertainty is the result of difficulties in interpretation of the various reported results of hearing-preservation surgery. Meaningful comparison between series is difficult because of the varied number of postoperative reporting criteria commonly in use today. Although it is acknowledged that the postoperative reporting criteria affect the hearing preservation rates, what is not readily appreciated is that preoperative selection criteria for hearing-preservation cases can also significantly affect the success rate of hearing-preservation acoustic neuroma surgery. This article models the many possible outcomes of hearing-preservation schwannoma surgery by use of the previously reported Cambridge series as an illustrative example. With these models some understanding can be gained of the effect of choosing various preoperative and postoperative hearing criteria on the overall hearing preservation success rate. PMID- 10471885 TI - Sinonasal lymphoma. AB - The clinical course of 49 patients with the diagnosis of non-Hodgkin's lymphoma were studied in the Department of Otolaryngology of the Hospital General de Mexico between December 1986 and March 1997. The most frequent clinical symptoms in these patients were: nasal obstruction, rhinorrhea, fever, weight loss, cervical lymph adenopathy, rhinopharyngeal tumor, ulceration on the palate and periorbital cellulitis. In 73% of the patients in this series the primary presentation was nasal. Sixty-six percent of the patients were classified as intermediate grade lymphoma according to the New Working Formulation, and 33% were in a I B state according to the Ann Arbor predominant immunotype was B cell in 63% of the cases. An extensive review of the literature is also presented. PMID- 10471884 TI - Relationship of human papillomavirus to ploidy in squamous cell carcinomas of the head and neck. AB - To establish the relationship between the presence of human papillomavirus (HPV) gene sequences and the development of genetic abnormalities, 31 squamous cell carcinomas of the head and neck were studied for the presence of HPV types 6b and 16 and the DNA content by flow cytometry. Eighteen (58%) cases were aneuploid. HPV DNA was present in seven (22.5%) tumors. Five of them were positive for the HPV type 6b and two for the HPV type 16. Aneuploidy was correlated with poorly differentiated tumors. No correlation was found between the presence of HPV, DNA content, or tumor differentiation. Consequently, the presence of HPV gene sequences does not seem to be related to a higher incidence of genetic abnormalities in squamous cell carcinomas of the head and neck. PMID- 10471886 TI - Temporal bone chondroblastoma. PMID- 10471887 TI - Benign fibrous histiocytoma of the floor of the mouth. PMID- 10471888 TI - Endoscopy during neurotomy of the nervus intermedius for geniculate neuralgia. PMID- 10471889 TI - T-cell non-Hodgkin's lymphoma of the larynx and hypopharynx. PMID- 10471890 TI - Anthropological approach to inform epidemiologic research on birth defects. PMID- 10471891 TI - Risks for FAS. PMID- 10471892 TI - Vasoactive teratogens and digital blood flow. PMID- 10471893 TI - Malrotation in conjunction with esophageal atresia/tracheo-esophageal fistula. AB - Esophageal atresia or tracheo-esophageal fistula (EA/TEF) often occurs in association with a well-defined group of other anomalies. We report the prevalence of malrotation and other intestinal anomalies in a large data series comprising 632 nontrisomic infants with EA/TEF ascertained by the California Birth Defects Monitoring Program from January 1, 1983 to December 31, 1994. Consistent with findings reported previously in smaller case series, our findings showed a notable prevalence of imperforate anus (9.0%) and duodenal atresia (5.2%), among other gastrointestinal defects. They also showed a previously unrecognized high prevalence of intestinal malrotation (4.4%). Compared with other infants studied, the infants with EA/TEF and malrotation of the intestine had a higher proportion of other associated anomalies (in particular intestinal, central nervous system, vertebral and rib, renal and genital anomalies). These findings indicate that intestinal malrotation is more common in infants with EA/TEF than is generally perceived, and that intestinal malrotation in an infant with EA/TEF is associated with a higher burden of additional congenital anomalies, suggesting that this group of infants may have more pervasive developmental deficits and poorer prognosis than has previously been recognized. PMID- 10471894 TI - Epidemiology of abdominal wall defects, Hawaii, 1986-1997. AB - The various types of abdominal wall defects are considered to differ in their etiologies, a hypothesis suggested by differences in their epidemiologies. This study examined the impact of selected demographic factors on abdominal wall defects (omphalocele, gastroschisis, and body stalk anomaly) included in a birth defects registry in Hawaii from 1986-1997. The total prevalence for the various defects were: omphalocele (2.76 per 10,000 births, 95% confidence interval (CI), 2.14-3.50), gastroschisis (3.01, 95% CI, 2. 36-3.77), and body stalk anomalies (0.32, 95% CI, 0.14-0.64). The prevalence increased over the 12-year period for both omphalocele (P = 0.052) and gastroschisis (P = 0.008). Women less than age 20 were at increased risk for a gastroschisis-affected pregnancy, while those age 40 and over were disproportionately more likely to have an omphalocele-affected pregnancy. Pacific Islanders had the lowest risk for omphalocele, whereas Far East Asians were least likely to have gastroschisis. Omphalocele rates were lower outside metropolitan Honolulu, while place of residence did not significantly impact gastroschisis risk. The 1-year survival rate was higher for gastroschisis than for omphalocele (88.5% and 70.7%, respectively), while none of the infants with body stalk anomalies was live-born. The results of this study tend to support the hypothesis of differing etiologies for the studied abdominal wall defects. PMID- 10471895 TI - Maternal periconceptional use of electric bed-heating devices and risk for neural tube defects and orofacial clefts. AB - Electric and magnetic fields are of concern as risk factors for adverse reproductive outcomes, including congenital anomalies. Among residential exposures to electric and magnetic fields, electric bed-heating devices such as electric blankets may be a substantial source of such exposures, and their use is fairly common. Two population-based case-control studies were analyzed to investigate whether the periconceptional use of electric blankets, bed warmers, or electrically heated waterbeds increased the risk of women to deliver infants or fetuses with neural tube defects (NTDs) or orofacial clefts. We obtained information on bed-heating devices from 538 NTD cases and their 539 controls in one study, and 265 NTD cases and 481 controls and 652 orofacial cleft cases and their 734 controls from another study. Our results revealed a few modestly elevated risks of certain anomaly phenotypes with maternal use of certain bed heating devices, but risks tended to be imprecise. In general, women who reported more frequent use of a bed-heating device, or longer duration of use, did not appear to have a higher risk for delivering offspring with anomalies than were women who reported less frequent or shorter-duration use. PMID- 10471896 TI - Proactive approach for the evaluation of fetal safety in chemical industries. AB - Women, their families, and employers are concerned about potential fetal risks that may be associated with occupational exposure to chemicals. In an attempt to quantify potential fetal risks in the petroleum industry, we conducted a literature review of selected chemical compounds to which Imperial Oil Limited (IOL) female personnel may be exposed. Medline, Toxline, and Dissertation Abstracts databases were utilized to search for all research papers published in any language from 1966-1996. Chemical exposures in these papers were compared to IOL chemical exposure indices from a specific refinery and chemical plant. In total, 559 studies obtained from the literature search related to the chemicals used in a specific refinery and chemical plant. Of these, only 21 studies explicitly stated some sort of exposure level for the various chemicals. Most of the selected female reproductive toxicology studies summarized explicitly stated chemical exposure levels: either as parts per million, stratifying as to number of days of exposure, or as estimates of the percentage of the threshold limit value. On comparing the occupational literature that presented either actual or estimated values of chemical exposure dose with the IOL routine rating factors in IOL's Products and Chemicals Divisions, we found that IOL chemical exposure levels overall were lower than those reported in the literature to be associated with fetal risks. A new proactive approach is presented to inform female workers and their families of the relative safety/risk of routine occupational exposures. This approach allows for the mitigation of the misperception of teratogenic risk and unjustified fears associated with it. PMID- 10471897 TI - Cerebrocortical microdysgenesis is enhanced in c57BL/6J mice exposed in utero to acetazolamide. AB - A small percentage of C57BL/6 mice spontaneously develop focal collections of neurons in the molecular layer of the cerebral neocortex. Usually only one "ectopia" is present in each affected brain. Studies in other mouse strains have shown that these ectopias occur before birth, probably because of a breach in the superficial glial membrane during neuronal migration. The ectopias are heritable and are caused by multiple genes. C57BL/6J mice exposed prenatally to acetazolamide, a carbonic anhydrase-specific inhibitor and teratogen, develop an increased frequency of limb malformations, especially in the right forelimb. In the present study, we hypothesized that the prevalence and severity of ectopias would be increased in acetazolamide-exposed mice because carbonic anhydrase plays a key role in brain development. Further, we wanted to determine whether there was a correlation between the side of limb deformity and the hemisphere containing an ectopia. Thus, we injected C57BL/6J time-mated mice intraperitoneally on embryonic day 9 with either sodium acetazolamide (750 mg/kg) or water. Histological analysis of the brains from 105 acetazolamide-exposed offspring and 89 control offspring revealed no difference in the overall prevalence of cerebrocortical ectopias between the acetazolamide and control groups: 34% of the acetazolamide-exposed and 28% of the control mice had ectopias. There was, however, a striking difference in the shape and size of ectopias: 67% of the ectopias were large in the acetazolamide-exposed group in comparison to 32% in controls. The acetazolamide-exposed offspring also were more likely to have multiple ectopias. Thus, there may be a genetic predisposition for developing ectopias in some mouse strains, but epigenetic factors such as prenatal exposure to acetazolamide can influence their severity. PMID- 10471898 TI - Intrauterine exposure to clomiphene and neonatal persistent hyperplastic primary vitreous. AB - A 4-year-and-10-month-old girl was diagnosed shortly after birth with persistent hyperplastic primary vitreous (PHPV). Her mother took clomiphene 100 mg daily for approximately 4 weeks and discontinued the drug once she had a positive pregnancy test. The exact time of gestation was not clear. Clomiphene is an estrogen antagonist effective in the treatment of anovulation. Various ocular side effects have been described in women taking the drug, including decreased vision, mydriasis, flashing lights, central scotoma, photophobia, diplopia, allergic reactions, retinal vasospasms, detachment posterior vitreous, and possibly posterior subcapsular cataracts. These occur in 1.5-10% of patients taking clomiphene. The potential effects of clomiphene on the fetus have been investigated in five animal studies. Cataracts were observed in fetal mice and rats, but not in monkeys. In humans, a case of congenital retinal aplasia was described. The possibility of clomiphene-induced congenital PHPV should be considered, especially in pregnant women who are taking a high and prolonged dose. PMID- 10471899 TI - Maternal smoking and craniosynostosis. AB - To investigate a possible association between maternal smoking during pregnancy and craniosynostosis (premature closure of one or more of the cranial sutures), a study was conducted using Swedish health registries. Infants with craniosynostosis (n = 304) without a known chromosome anomaly were selected among 1,413,811 infants born between 1983-1996 with known smoking exposure in early pregnancy. A statistically significant association between maternal smoking and craniosynostosis was found (adjusted odds ratio (OR), 1.45; 95% confidence interval (CI), 1.13-1.87), but this association was only valid for isolated defects (OR, 1.67; 95% CI, 1.27-2.19). For associated craniosynostosis (malformation syndromes included), a negative (nonsignificant) association with maternal smoking was indicated instead. For isolated craniosynostosis (all types), a dose-dependent effect of maternal smoking was indicated (OR and 95% CI for smoking <10 cigarettes/day, 1.45 (1.04-2.02); OR and 95% CI for smoking >/=10 cigarettes/day, 2.12 (1.50-2.99)), but was not statistically significant. Among the different types of craniosynostosis, premature closure of the sagittal suture showed the strongest association with maternal smoking (OR, 1.48; 95% CI, 1. 02 2.14), whereas for coronal suture defects, no association with maternal smoking could be detected (OR, 1.02; 95% CI, 0.47-2.21). As no obvious confounders were detected, the present study supports an earlier report of an association between maternal smoking during pregnancy and at least some types of craniosynostosis. PMID- 10471901 TI - Teratogen update: paternal exposures-reproductive risks. PMID- 10471900 TI - Role of free radicals in the limb teratogenicity of L-NAME (N(G)-nitro-(L) arginine methyl ester): a new mechanistic model of vascular disruption. AB - In continuing studies of limb effects resulting from fetal exposure to N(G)-nitro (L)-arginine methyl ester (L-NAME), we examined the early time course of vascular changes and the effectiveness of fetal intraamniotic injection. Vascular engorgement and hemorrhage occurred within 4 hr of L-NAME treatment on gestational day (gd) 17, and direct injection appeared to be as effective as maternal intraperitoneal injection in inducing limb hemorrhage. Further studies examined protein nitration and electron transport inhibition in tissues of exposed fetuses. L-NAME caused significant increases in nitrotyrosine (NT) formation in limb but not in heart or brain, and reduced electron transport rates in limb. Three agents, alpha-phenyl-N-t-butylnitrone (PBN), a radical trap and inhibitor of inducible nitric oxide synthase (iNOS), allopurinol, an inhibitor of xanthine oxidase, and aminoguanidine, a relatively specific inhibitor of iNOS, significantly moderated limb hemorrhage and protein nitration in distal limb. These results suggest that L-NAME works directly on the fetal limb vasculature and indicate a cytotoxic role for peroxynitrite, a potent oxidant and nitrating agent that is the reaction product of nitric oxide and superoxide anion radical. We propose that L-NAME and other vasoactive toxicants disrupt the fetal limb in a sequential process. Initially, nitric oxide (NO) is depleted, causing hemorrhage and edema in the limb. Within hours, iNOS is induced, resulting in cytotoxic tissue concentrations of NO and reactive nitrogen species that induce apoptosis and/or necrosis in the limb. We suggest that L-NAME exposure may serve as a model of vascular disruptive limb malformations. PMID- 10471902 TI - A golden bowl of rice. PMID- 10471903 TI - False reports and the ears of men. PMID- 10471904 TI - In search of horizontal gene transfer. PMID- 10471905 TI - The drug discovery factory: An inevitable evolutionary consequence of high throughput parallel processing. PMID- 10471906 TI - The essential prerequisites for quantitative RT-PCR. PMID- 10471907 TI - Reply to "GM gene flow" PMID- 10471908 TI - Baby food makers to drop GM ingredients PMID- 10471909 TI - Genentech's 2nd IPO PMID- 10471910 TI - Aptamers on the scaffold PMID- 10471911 TI - Topical gene delivery PMID- 10471912 TI - UH researcher files transgenesis suit. University of Hawaii. PMID- 10471913 TI - Europe edges closer to GMO patent harmonization. PMID- 10471914 TI - ArQule-Pfizer deal breaks mold. PMID- 10471915 TI - Johnson & Johnson acquires Centocor. PMID- 10471916 TI - World Bank task force push/pulls AIDS vaccines. PMID- 10471917 TI - US food labeling policy softens. PMID- 10471918 TI - Monsanto remains a magnet for GM opposition. PMID- 10471919 TI - Nuclear medicine finds the right chemistry. PMID- 10471920 TI - Pathways for neural stem cell biology and repair. PMID- 10471921 TI - Budding actors in mammalian G-protein signaling. PMID- 10471923 TI - New tools for chloroplast genetic engineering. PMID- 10471922 TI - Fine-tuning an engineered intein. PMID- 10471924 TI - The greening of chimeric oligonucleotides. PMID- 10471925 TI - Drug screening--beyond the bottleneck. AB - High-throughput screening still has a long way to go if it is to achieve its often-touted potential. PMID- 10471926 TI - Neuron-restrictive silencer elements mediate neuron specificity of adenoviral gene expression. AB - Neuron-restrictive silencer elements (NRSEs) were used to target the gene expression of adenoviral vectors specifically to neuron cells in the central nervous system. By generating adenoviral constructs in which NRSE sequences were placed upstream from the ubiquitous phosphoglycerate kinase promoter, the specificity of expression of a luciferase reporter gene was tested in both cell lines and primary cultures. Whereas transgene expression was negligible in nonneuronal cells following infection with an adenovirus containing 12 NRSEs, neuronal cells strongly expressed luciferase when infected with the same adenovirus. The NRSEs restricted expression of the luciferase gene to neuronal cells in vivo when adenoviruses were injected both intramuscularly into mice and intracerebrally into rats. This NRSE strategy may avoid side effects resulting from the ectopic expression of therapeutic genes in the treatment of neurological diseases. In particular, it may allow the direct transfection of motor neurons without promoting transgene expression within inoculated muscles or the secretion of transgene products into the bloodstream. PMID- 10471927 TI - Immunization via hair follicles by topical application of naked DNA to normal skin. AB - In order to test the immune response generated to small amounts of foreign protein in skin, we applied naked DNA in aqueous solution to untreated normal skin. Topical application of plasmid expression vectors for lacZ and the hepatitis B surface antigen (HBsAg) to intact skin induced antigen-specific immune responses that displayed TH2 features. For HBsAg, specific antibody and cellular responses were induced to the same order of magnitude as those produced by intramuscular injection of the commercially available recombinant HBsAg polypeptide vaccine. Finally, topical gene transfer was dependent on the presence of normal hair follicles. PMID- 10471928 TI - Nuclear targeting peptide scaffolds for lipofection of nondividing mammalian cells. AB - Lipofection of nondividing cells is inefficient because much of the transfected DNA is retained in endosomes, and that which escapes to the cytoplasm enters the nucleus at low rates. To improve the final rate-limiting step of nuclear import, we conjugated a nonclassical nuclear localization signal (NLS) containing the M9 sequence of heterogeneous nuclear ribonucleoprotein (hnRNP) A1, to a cationic peptide scaffold derived from a scrambled sequence of the SV40 T-antigen consensus NLS (ScT). The ScT was added to improve DNA binding of the M9 sequence. Lipofection of confluent endothelium with plasmid complexed with the M9-ScT conjugate resulted in 83% transfection and a 63-fold increase in marker gene expression. The M9-ScT conjugate localized fluorescent plasmid into the nucleus of permeabilized cells, and addition of the nuclear pore blocker wheat germ agglutinin prevented nuclear import. This method of gene transfer may lead to viral- and lipid-free transfection of nondividing cells. PMID- 10471929 TI - Genetic screens in yeast to identify mammalian nonreceptor modulators of G protein signaling. AB - We describe genetic screens in Saccharomyces cerevisiae designed to identify mammalian nonreceptor modulators of G-protein signaling pathways. Strains lacking a pheromone-responsive G-protein coupled receptor and expressing a mammalian yeast Galpha hybrid protein were made conditional for growth upon either pheromone pathway activation (activator screen) or pheromone pathway inactivation (inhibitor screen). Mammalian cDNAs that conferred plasmid-dependent growth under restrictive conditions were identified. One of the cDNAs identified from the activator screen, a human Ras-related G protein that we term AGS1 (for activator of G-protein signaling), appears to function by facilitating guanosine triphosphate (GTP) exchange on the heterotrimeric Galpha. A cDNA product identified from the inhibitor screen encodes a previously identified regulator of G-protein signaling, human RGS5. PMID- 10471930 TI - Rapid identification of DNA-binding proteins by mass spectrometry. AB - We report a protocol for the rapid identification of DNA-binding proteins. Immobilized DNA probes harboring a specific sequence motif are incubated with cell or nuclear extract. Proteins are analyzed directly off the solid support by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The determined molecular masses are often sufficient for identification. If not, the proteins are subjected to mass spectrometric peptide mapping followed by database searches. Apart from protein identification, the protocol also yields information on posttranslational modifications. The protocol was validated by the identification of known prokaryotic and eukaryotic DNA-binding proteins, and its use provided evidence that poly(ADP-ribose) polymerase exhibits DNA sequence specific binding to DNA. PMID- 10471931 TI - A genetic system yields self-cleaving inteins for bioseparations. AB - A self-cleaving element for use in bioseparations has been derived from a naturally occurring, 43 kDa protein splicing element (intein) through a combination of protein engineering and random mutagenesis. A mini-intein (18 kDa) previously engineered for reduced size had compromised activity and was therefore subjected to random mutagenesis and genetic selection. In one selection a mini intein was isolated with restored splicing activity, while in another, a mutant was isolated with enhanced, pH-sensitive C-terminal cleavage activity. The enhanced-cleavage mutant has utility in affinity fusion-based protein purification. These mutants also provide new insights into the structural and functional roles of some conserved residues in protein splicing. PMID- 10471932 TI - DNA shuffling of subgenomic sequences of subtilisin. AB - DNA family shuffling of 26 protease genes was used to create a library of chimeric proteases that was screened for four distinct enzymatic properties. Multiple clones were identified that were significantly improved over any of the parental enzymes for each individual property. Family shuffling, also known as molecular breeding, efficiently created all of the combinations of parental properties, producing a great diversity of property combinations in the progeny enzymes. Thus, molecular breeding, like classical breeding, is a powerful tool for recombining existing diversity to tailor biological systems for multiple functional parameters. PMID- 10471933 TI - Stable one-step technetium-99m labeling of His-tagged recombinant proteins with a novel Tc(I)-carbonyl complex. AB - We have developed a technetium labeling technology based on a new organometallic chemistry, which involves simple mixing of the novel reagent, a 99m Tc(I) carbonyl compound, with a His-tagged recombinant protein. This method obviates the labeling of unpaired engineered cysteines, which frequently create problems in large-scale expression and storage of disulfide-containing proteins. In this study, we labeled antibody single-chain Fv fragments to high specific activities (90 mCi/mg), and the label was very stable to serum and all other challenges tested. The pharmacokinetic characteristics were indistinguishable from iodinated scFv fragments, and thus scFV fragments labeled by the new method will be suitable for biodistribution studies. This novel labeling method should be applicable not only to diagnostic imaging with 99mTc, but also to radioimmunotherapy approaches with 186/188 Re, and its use can be easily extended to almost any recombinant protein or synthetic peptide. PMID- 10471934 TI - Antibody recognition imaging by force microscopy. AB - We have developed a method that combines dynamic force microscopy with the simultaneous molecular recognition of an antigen by an antibody, during imaging. A magnetically oscillated atomic force microscopy tip carrying a tethered antibody was scanned over a surface to which lysozyme was bound. By oscillating the probe at an amplitude of only a few nanometers, the antibody was kept in close proximity to the surface, allowing fast and efficient antigen recognition and gentle interaction between tip and sample. Antigenic sites were evident from reduction of the oscillation amplitude, as a result of antibody-antigen recognition during the lateral scan. Lysozyme molecules bound to the surface were recognized by the antibody on the scanning tip with a few nanometers lateral resolution. In principle, any ligand can be tethered to the tip; thus, this technique could potentially be used for nanometer-scale epitope mapping of biomolecules and localizing receptor sites during biological processes. PMID- 10471935 TI - A galinstan expansion femtosyringe for microinjection of eukaryotic organelles and prokaryotes. AB - A galinstan expansion femtosyringe enables femtoliter to attoliter samples to be introduced into prokaryotes and subcellular compartments of eukaryotes. The method uses heat-induced expansion of galinstan (a liquid metal alloy of gallium, indium, and tin) within a glass syringe to expel samples through a tip diameter of about 0.1 microm. The narrow tip inflicts less damage than conventional capillaries, and the heat-induced expansion of the galinstan allows fine control over the rate of injection. We demonstrate injection of Lucifer Yellow and Lucifer Yellow-dextran conjugates into cyanobacteria, and into nuclei and chloroplasts of higher organisms. Injection of a plasmid containing the bla gene into the cyanobacterium Phormidium laminosum resulted in transformed ampicillin resistant cultures. Green fluorescent protein was expressed in attached leaves of tobacco and Vicia faba following injection of DNA containing its gene into individual chloroplasts. PMID- 10471936 TI - Fluorescent antibiotic resistance marker for tracking plastid transformation in higher plants. AB - Plastid transformation in higher plants is accomplished through a gradual process, during which all the 300-10,000 plastid genome copies are uniformly altered. Antibiotic resistance genes incorporated in the plastid genome facilitate maintenance of transplastomes during this process. Given the high number of plastid genome copies in a cell, transformation unavoidably yields chimeric tissues, which requires the identification of transplastomic cells in order to regenerate plants. In the chimeric tissue, however, antibiotic resistance is not cell autonomous: transplastomic and wild-type sectors both have a resistant phenotype because of phenotypic masking by the transgenic cells. We report a system of marker genes for plastid transformation, termed FLARE-S, which is obtained by translationally fusing aminoglycoside 3"-adenyltransferase with the Aequorea victoria green fluorescent protein. 3"-adenyltransferase (FLARE-S) confers resistance to both spectinomycin and streptomycin. The utility of FLARE-S is shown by tracking segregation of individual transformed and wild-type plastids in tobacco and rice plants after bombardment with FLARE-S vector DNA and selection for spectinomycin and streptomycin resistance, respectively. This method facilitates the extension of plastid transformation to nongreen plastids in embryogenic cells of cereal crops. PMID- 10471937 TI - Inducible isopentenyl transferase as a high-efficiency marker for plant transformation. AB - Overexpression of the isopentenyltransferase gene (ipt) from the Ti-plasmid of Agrobacterium tumefaciens increases cytokinin levels, leading to generation of shoots from transformed plant cells. When combined with a dexamethasone-inducible system for controlling expression, ipt expression can be used to select for transgenic regenerants without using an antibiotic-resistance marker. The combined system allows efficient cointroduction of multiple genes (in addition to ipt) and produces transgenic plants without morphological or developmental defects. PMID- 10471939 TI - Recent patents in gene transfer PMID- 10471938 TI - New rules for natural products research. PMID- 10471941 TI - People PMID- 10471940 TI - The philosopher as biologist PMID- 10471942 TI - New products PMID- 10471943 TI - Cardiovascular disease drug discovery. PMID- 10471944 TI - Mass spectrometry of nucleic acids. PMID- 10471945 TI - Make biology compulsory for presidential candidates. PMID- 10471946 TI - GM gene flow. PMID- 10471948 TI - Reply to "GM food labelling" PMID- 10471947 TI - GM food labeling. PMID- 10471950 TI - GM roundup PMID- 10471949 TI - Japan to label GMOs PMID- 10471951 TI - USDA aims at africa PMID- 10471952 TI - It's the real thing PMID- 10471953 TI - Cartoon PMID- 10471955 TI - AAV muscles ahead PMID- 10471954 TI - CT turns focus to StemCell PMID- 10471956 TI - A spoon full of honey PMID- 10471957 TI - Research collaborations PMID- 10471958 TI - Retargeting tropism PMID- 10471959 TI - In silico matchmaking PMID- 10471960 TI - Influenza from scratch PMID- 10471961 TI - Breeding molecules PMID- 10471963 TI - Imaging antigens by force PMID- 10471962 TI - Silence is golden PMID- 10471964 TI - Human genes find function in yeast PMID- 10471965 TI - Gene transfer gets a boost PMID- 10471966 TI - Mass attack on proteins PMID- 10471968 TI - One-step radiolabeling of scFvs PMID- 10471967 TI - Markers outgrow antibiotic resistance PMID- 10471969 TI - Rationalizing a random approach PMID- 10471971 TI - Lighting up transplastomics PMID- 10471972 TI - Director bids aloha, storm brews over UH NSF institute. University of Hawaii. PMID- 10471970 TI - Plastid microinjection PMID- 10471973 TI - Treatment of an autoimmune disease with "classical" T cell veto: a proposal. AB - Immune responses protect against infectious diseases and cancers. In normal circumstances, the immune system is tolerant to self. However, under certain conditions this tolerance is broken. The immune system attacks otherwise normal tissue. An autoimmune disease ensues. Strategies are now being sought that remove the pathogenic T cells without affecting other immune functions. "Classical" veto has been described as an immune suppressive mechanism able to remove T cells in a highly specific and effective manner. The present article briefly reviews the current knowledge on the development of autoreactive T cells and their regulation in the periphery. It describes "classical" veto, its mechanisms, and its novel applications. Finally, it argues that "classical" veto can be adapted to treat an autoimmune disease, such as type I diabetes mellitus. PMID- 10471975 TI - The importance of specific IgG and IgE autoantibodies to retinal S antigen, total serum IgE, and sCD23 levels in autoimmune and infectious uveitis. AB - Autoimmunity plays an important role in the development of uveitis. The uveitis are linked to Th1 or Th2 lymphocyte activation. We studied 41 patients with uveitis, divided into autoimmune uveitis (n = 32) and infectious uveitis (n = 9), 30 normal controls, and 20 asthmatic atopic without ocular diseases. The infectious uveitis patients were separated into bacterial (n = 6) and toxoplasmic (n = 3) retinochoroiditis. We measured IgE and sCD23 serum levels and specific IgG and IgE to retinal S antigen by ELISA tests. The IgE levels were 500 +/- 325 kU/L in autoimmune uveitis, 57 +/- 35 kU/L in bacterial uveitis, 280 +/- 38 kU/L in toxoplasmic retinochoroiditis, 75 +/- 32 kU/L in the controls, and 557 +/- 243 kU/L in atopics (P < 0.0005). The sCD23 levels were 10.4 +/- 5.4 ng/ml in autoimmune uveitis, 3.7 +/- 1.17 ng/ml in bacterial uveitis, 6.76 +/- 1.36 ng/ml in toxoplasmic retinochoroiditis, 3.4 +/- 1 ng/ml in controls, and 8.35 +/- 2.2 ng/ml in atopic patients (P < 0.005). The specific IgG to retinal S antigen was positive in 27 of 32 cases, and the specific IgE to retinal S antigen was positive in 22 of 32 autoimmune uveitis. The bacterial uveitis patients as well as the controls were negative for both autoantibodies to retinal S antigen. The toxoplasmic retinochoroiditis patients presented specific IgG and IgE to retinal S antigen in two of three cases, respectively, one of them with overlap of both antibodies. These results suggest the importance of specific IgG and IgE to retinal S antigen in autoimmune uveitis, which, along with higher IgE and sCD23 levels, reveal Th2 activation. PMID- 10471974 TI - New insights into the biology of the acute phase response. AB - Innate or natural immunity is a highly conserved defense mechanism against infection found in all multicellular organisms. The acute phase response is the set of immediate inflammatory responses initiated by pattern recognition molecules. These germ cell-encoded proteins recognize microbial pathogens based on shared molecular structures and induce host responses that localize the spread of infection and enhance systemic resistance to infection. Innate immunity also influences the initiation and type of adaptive immune response by regulating T cell costimulatory activity and antigen presentation by antigen presenting cells and by influencing mediator production, which affects lymphocyte function and trafficking. Acute phase protein concentrations rapidly increase after infection, and their production is controlled primarily by IL-6- and IL-1-type cytokines. The acute phase proteins provide enhanced protection against microorganisms and modify inflammatory responses by effects on cell trafficking and mediator release. For example, serum amyloid A has potent leukocyte activating functions including induction of chemotaxis, enhancement of leukocyte adhesion to endothelial cells, and increased phagocytosis. The constellation of inflammatory responses seen after endotoxin administration to humans represents an in vivo model of the acute phase response. Studies with inflammatory modifying agents, such as soluble dimeric TNF receptor and IL-10, show that these responses are not dependent on a single mediator but result from multiple overlapping inflammatory pathways. Understanding the factors that initiate and alter the magnitude and duration of the acute phase response represents an important step in the development of new therapies for infectious and inflammatory diseases. PMID- 10471976 TI - Intracerebral production of tumor necrosis factor-alpha, a local neuroprotective agent, in Alzheimer disease and vascular dementia. AB - The local pattern of proinflammatory cytokine release was studied in Alzheimer disease (AD) and vascular dementia (VAD), by measuring intrathecal levels of IL-1 beta, IL-6, TNF-alpha, and its naturally occurring antagonists, soluble TNF receptors I and II. The cytokine levels were related to neuronal damage, as measured by the intrathecal tau concentration, to cerebral apoptosis assessed by levels of Fas/APO-1 and bcl-2, and to clinical variables. In vitro analysis was performed to study the effect of TNF-alpha on the production of bcl-2, an antiapoptotic factor, by human neuronal cells. Patients with both AD and VAD displayed significantly higher intrathecal levels of TNF-alpha compared to controls. In addition, patients with AD showed significantly negative correlations between the intrathecal levels of TNF-alpha and the levels of Fas/APO-1 as well as of tau protein. The level of bcl-2 in supernatants of TNF alpha-exposed cultures of human neuronal cells was up to three times higher than in control supernatants. Our study demonstrates intrathecal production of TNF alpha in patients with dementias, suggesting that this cytokine may have a neuroprotective role in these neurodegenerative conditions as evidenced by negative correlations between this cytokine and (i) levels of intrathecal Fas/APO 1 and (ii) levels of tau protein, both parameters closely related to brain damage. Our in vitro data suggest that TNF-alpha exerts its neuroprotective effect by stimulating neuronal cells to express bcl-2, a molecule which downregulates apoptosis. PMID- 10471978 TI - Activation of human peripheral blood T cells does not lead to increased P glycoprotein expression. AB - The expression of P-glycoprotein (Pgp) on normal human lymphocytes, and its drug exclusion capacity, implies that Pgp might be involved in cytokine secretion. We used two-color flow cytometry to detect simultaneously Pgp expression and IL-2 accumulation in resting and mitogen-activated human lymphocytes. Among resting lymphocytes from five healthy donors less than 1% were Pgp+ as determined by reactivity with the anti-Pgp monoclonal antibody (mAb) 4E3. The percentage of Pgp+ lymphocytes increased to 3% after 24 hr of mitogenic stimulation that induced maximal production of cytoplasmic IL-2. The percentage of lymphocytes that coexpressed membrane Pgp and cytoplasmic IL-2 accounted for < 10% of the total IL-2 producing lymphocytes. Finally, mitogen-induced cytoplasmic IL-2 accumulation was enhanced by stimulation in the presence of monensin but not the Pgp functional inhibitor verapamil. Because mAb 4E3 detected lower than expected numbers of Pgp+ lymphocytes, we compared the binding of mAbs MRK16 and 4E3 concomitant with doxorubicin (DOX)-uptake by K562 and R7 tumor cells and purified CD8+ lymphocytes. The MRK16 mAb was found to be sensitive but not very specific (30%). In contrast, the sensitivity of 4E3 was equivalent to MRK16 (98%) and was highly specific (98.5%). There was also a positive association between DOX efflux and the level of Pgp expression as detected by 4E3 but not MRK16. Thus, human T cells do not markedly up-regulate their expression of functional Pgp molecules as detected by mAb 4E3 following activation, suggesting that Pgp does not play a major role in IL-2 secretion by activated T cells. PMID- 10471979 TI - Microbial biofilms: their development and significance for medical device-related infections. AB - Microbial adhesion and biofilm formation on medical devices represent a common occurrence that can lead to serious illness and death. The process by which bacteria and yeast colonize open and closed implants is fairly complicated and involves a series of steps commencing with deposition of host substances onto the material. Prevention and treatment of established biofilms with antimicrobial agents are difficult because the organisms are encased within a protected microenvironment. Efforts to reduce adhesion using specially developed materials, such as hydrophilic or heparin coated, have had modest success once applied to the patient. The reason, at least for the most part, is the diverse milieu into which devices are placed and the multitude of ways in which organisms can colonize surfaces. A better understanding of the process is required, and the knowledge gained must be used to devise new strategies as alternatives to the traditional employment of antibiotics. These new approaches may still use antibiotics but at different concentrations (low to prevent and high to treat infection) and in a different manner (perhaps spiked therapy in which there is a delay between doses to reduce the risk of drug resistance and impact on normal flora). The possibility of applying functional foods to patient management should also be pursued. PMID- 10471980 TI - FDA evaluations using in vitro metabolism to predict and interpret in vivo metabolic drug-drug interactions: impact on labeling. AB - Recent advances in in vitro metabolism methods have led to an improved ability to predict clinically relevant metabolic drug-drug interactions. To address the relationships of in vitro metabolism data and in vivo metabolism outcomes, the Office of Clinical Pharmacology and Biopharmaceutics in the Center for Drug Evaluation and Research, Food and Drug Administration, evaluated a number of recently approved new drug applications. The goal of these evaluations was to determine the contribution of in vitro metabolism data in (1) predicting in vivo drug-drug interactions, (2) determining the need to conduct an in vivo drug-drug interaction study, and (3) incorporating findings into drug product labeling. Ten cases are presented in this article. They fall into two major groups: (1) in vitro data were predictive of in vivo results, and (2) in vitro data were not predictive of in vivo results. Discussion of these cases highlights factors limiting predictability of in vivo metabolic interactions from in vitro metabolism data. The integration of these findings into drug product labeling is also discussed. PMID- 10471981 TI - Effect of meal timing not critical for the pharmacokinetics of tegaserod (HTF 919). AB - This study assessed the pharmacokinetic profiles of administering tegaserod (HTF 919) at different time intervals with respect to a meal. It was a randomized, open-label, two-phase, five-period crossover study. In the first phase, 18 healthy subjects received a single 12 mg oral dose of tegaserod administered either 30 or 15 minutes prior to the start of the 600-calorie, fat-rich breakfast. In the second phase, subjects received a single 12 mg oral dose of tegaserod 1 minute before, 2.5 hours after the start of meal, or with a continued 4-hour postdose fast. Safety assessment and plasma samples for the determination of drug concentration were obtained for 24 hours postdose. Noncompartmental analysis results indicated that the AUC of tegaserod was reduced by almost half under fed conditions compared to the fasted condition. Exploratory analyses were implemented to further investigate the absorption characteristics of tegaserod under different fed conditions. A numerical deconvolution approach was used to obtain the tegaserod oral absorption versus time profiles under both fasted and fed conditions. The tegaserod oral absorption versus time profiles were then fitted by NONMEM to a model containing two absorption phases. Based on the absorption analyses, we found that the reduction in the bioavailability of tegaserod under fed conditions was primarily due to a decrease in the extent of absorption and less so to a decrease in the absorption rate(s). Therefore, although the timing of administration of food does not appear to significantly alter the pharmacokinetics of tegaserod, the administration of food reduces the AUC by approximately 50%. PMID- 10471977 TI - Activation of mononuclear cells by interleukin-12: an in vivo study in chimpanzees. AB - Interleukin (IL)-12 is considered a central regulator of host resistance against a variety of pathogens. Therefore, IL-12 has been advocated as a potential therapeutic agent in infections. To determine the in vivo effects of IL-12 on mononuclear cells involved in the host immune response, four chimpanzees received an intravenous injection of recombinant IL-12 (1 microgram/kg). IL-12 induced a sustained decrease in lymphocyte counts, with decreases in CD3+/CD4+ and CD3+/CD8+ cells, while monocyte counts showed a transient increase. IL-12 injection resulted in a shift toward a Th1-mediated immune response as indicated by increased interferon-gamma production during whole-blood stimulation, while not influencing IL-4 production. IL-12-induced activation of NK cells and phagocytes, as indicated by increased NK cell cytotoxicity and increased plasma levels of granzymes A and B and of chitotriosidase activity. These data support the hypothesis that IL-12 may serve as a useful therapeutic agent in infections where a cell-mediated response is protective. PMID- 10471982 TI - Steady-state pharmacokinetics and dose proportionality of troglitazone and its metabolites. AB - This study evaluated the steady-state pharmacokinetics and dose proportionality of troglitazone, metabolite 1 (sulfate conjugate), and metabolite 3 (quinone metabolite) following administration of daily oral doses of 200, 400, and 600 mg troglitazone for 7 days (per dosing period) to 21 subjects. During each dosing period, plasma samples were collected predose on days 1, 5, 6 and 7 and serially for 24 hours on day 7. Steady-state plasma concentrations for troglitazone, metabolite 1, and metabolite 3 were achieved by day 7. Troglitazone was rapidly absorbed with mean tmax values of 2.7 to 2.9 hours. Mean Cmax and AUC(0-24) values for troglitazone, metabolite 1, and metabolite 3 increased proportionally with increasing troglitazone doses over the clinical dose range of 200 mg to 600 mg administered once daily. Mean troglitazone CL/F, percent fluctuation, and AUC ratios of metabolite 1 and metabolite 3 to troglitazone were similar across dose groups. These data suggest that the pharmacokinetics and disposition of troglitazone and its metabolites are independent of dose over the dose range studied. Thus, troglitazone, metabolite 1, and metabolite 3 displayed linear pharmacokinetics at steady-state. PMID- 10471983 TI - Rabeprazole: pharmacokinetics and tolerability in patients with stable, end-stage renal failure. AB - The authors compare the pharmacokinetic profiles, safety, and tolerability of rabeprazole, a new proton pump inhibitor (PPI), in healthy volunteers and in subjects with stable, end-stage renal failure. This single-center, open-label trial included two groups of subjects: 10 healthy males with 24-hour creatinine clearance > or = 90 mL/min/m2 and 10 males with renal failure (24-hour creatinine clearance < or = 5 mL/min/m2) receiving hemodialytic therapy. Normal subjects received a single, oral 20 mg rabeprazole dose. Those with renal failure received a 20 mg dose of rabeprazole on the day after hemodialysis and a second dose after a 2-week washout period during dialysis. Blood samples were drawn before and up to 24 hours after rabeprazole administration for determination of plasma rabeprazole concentrations by high-performance liquid chromatography. Safety and tolerability of rabeprazole were determined by reporting adverse events and comparing vital signs, ECG, physical examinations, and clinical laboratory tests before and during treatment. Comparison of pharmacokinetic results from healthy volunteers with those from subjects with renal failure indicated no clinically significant differences between groups. In addition, there were no statistically significant differences between any pharmacokinetic parameters recorded during or after hemodialysis. Rabeprazole was well tolerated by both groups. Only two drug related adverse events were reported, and there were no significant treatment emergent changes in vital signs or ECG. Treatment-emergent changes in hematologic and clinical chemistry parameters were observed for a few subjects in each group and generally represented only slight deviations from the normal range. These results indicate that no dosage adjustment of rabeprazole is required in patients with renal dysfunction. These findings and the well-documented clinical efficacy of this new PPI in patients with gastric ulcers, duodenal ulcers, or gastroesophageal reflux disease support rabeprazole's use in the treatment of patients with acid peptic disorders. PMID- 10471984 TI - Gabapentin does not affect antipyrine clearance. AB - The effect of gabapentin on antipyrine clearance was assessed in 12 healthy male volunteers, using a known enzyme inducer, phenytoin, as control. Subjects received gabapentin 400 mg or phenytoin 100 mg three times daily for 2 weeks. Antipyrine tests were performed before, during, and after treatment with gabapentin or phenytoin. In contrast to phenytoin, chronic administration of gabapentin did not affect antipyrine clearance. Gabapentin appears to have little potential for drug interactions. PMID- 10471985 TI - Inhibition of caffeine metabolism by estrogen replacement therapy in postmenopausal women. AB - This study was conducted to investigate the effect of therapeutic estrogen on cytochrome P450 1A2-mediated metabolism in postmenopausal women using caffeine as a model substrate. Twelve healthy postmenopausal women underwent estrogen replacement therapy in the form of estradiol (Estrace). Estradiol was initiated at a dose of 0.5 mg a day and titrated to achieve a steady-state plasma concentration of 50 to 150 pg/ml. Caffeine metabolic ratios (CMR; paraxanthine/caffeine) were assessed both before and after 8 weeks of estrogen replacement. For the 12 subjects, there was a mean reduction in CMR of -29.2 +/- 25.0 (p = 0.0019). Consistent with previous results found in younger women, these results indicate that exogenous estrogen in older women may inhibit CYP1A2 mediated caffeine metabolism. PMID- 10471986 TI - Effect of multiple doses of montelukast, a CysLT1 receptor antagonist, on digoxin pharmacokinetics in healthy volunteers. AB - The effect of multiple oral doses of montelukast, a cysLT1 receptor antagonist, on the pharmacokinetics of oral digoxin was studied in healthy male volunteers in a randomized double-blind two-period crossover study. Subjects received 10 mg of montelukast or placebo daily for 11 days. On day 7, they received a single 0.5 mg oral dose of digoxin elixir. The pharmacokinetic parameters of digoxin (AUC0- >24' AUC0-->infinity' Cmax' tmax' t1/2) and cumulative urinary excretion over 120 hours were not affected by the multiple doses of montelukast. The 90% confidence interval for each of these parameters fell within prespecified clinically acceptable bounds. Side effects were mild and transient. This suggests that concurrent administration of montelukast and digoxin was well tolerated. Concurrent treatment with montelukast has no effect on the pharmacokinetics of digoxin. PMID- 10471987 TI - Lack of pharmacokinetic interaction between oral pantoprazole and cisapride in healthy adults. AB - Pantoprazole, an irreversible proton pump inhibitor, may be administered with cisapride, a prokinetic agent. As increased cisapride concentrations may result in longer electrocardiogram (ECG) QTc intervals, a crossover study was conducted in healthy subjects to evaluate the oral pharmacokinetic interaction between cisapride (20 mg) and pantoprazole (40 mg). After dosing, serial blood samples and 12-lead ECGs were collected, and cisapride plasma concentrations were quantitated. For cisapride alone, mean parameter values were the following: peak concentration (Cmax), 56 ng/mL; time to Cmax (tmax), 1.7 hours; area under the concentration-time curve (AUC), 426 ng x h/mL; and terminal half-life (t1/2), 5.8 hours. Pantoprazole coadministration did not alter cisapride AUC or other pharmacokinetic parameters except for a slight 17% decrease in Cmax' resulting in 90% confidence limits of 79% to 88%, which were marginally outside strict bioequivalence limits. In addition, cisapride did not affect ECG QTc intervals, with or without pantoprazole. Therefore, no dosage adjustment is needed when pantoprazole and cisapride are coadministered. PMID- 10471988 TI - Myosins and deafness. AB - The discovery in the past few years of a huge diversity within the myosin superfamily has been coupled with an understanding of the role of these motor proteins in various cellular functions. Extensive studies have revealed that myosin isoforms are not only involved in muscle contraction but also in crucial functions of many specialized mammalian cells such as melanocytes, kidney and intestinal brush border microvilli, nerve growth cones or inner ear hair cells. A search for genes involved in the pathology of human genetic deafness resulted in identification of three novel myosins: myosin VI, myosin VIIA and, very recently, myosin XV. The structure, tissue and cellular distribution of these myosin isoforms, as well as mutations detected within their genes that have been found to affect the hearing process, are described in this review. PMID- 10471989 TI - Developmental changes in the activation properties and ultrastructure of fast- and slow-twitch muscles from fetal sheep. AB - At early stages of muscle development, skeletal muscles contract and relax slowly, regardless of whether they are destined to become fast- or slow-twitch. In this study, we have characterised the activation profiles of developing fast- and slow-twitch muscles from a precocial species, the sheep, to determine if the activation profiles of the muscles are characteristically slow when both the fast and slow-twitch muscles have slow isometric contraction profiles. Single skinned muscle fibres from the fast-twitch flexor digitorum longus (FDL) and slow-twitch soleus muscles from fetal (gestational ages 70, 90, 120 and 140 days; term 147 days) and neonatal (8 weeks old) sheep were used to determine the isometric force pCa (pCa = -log10[Ca2+]) and force-pSr relations during development. Fast-twitch mammalian muscles generally have a greatly different sensitivity to Ca2+ and Sr2+ whereas slow-twitch muscles have a similar sensitivity to these divalent cations. At all ages studied, the force-pCa and force-pSr relations of the FDL muscle were widely separated. The mean separation of the mid-point of the curves (pCa50 pSr50) was approximately 1.1. This is typical of adult fast-twitch muscle. The force-pCa and force-pSr curves for soleus muscle were also widely separated at 70 and 90 days gestation (pCa50-pSr50 approximately 0.75); between 90 days and 140 days this separation decreased significantly to approximately 0.2. This leads to a paradoxical situation whereby at early stages of muscle development the fast muscles have contraction dynamics of slow muscles but the slow muscles have activation profiles more characteristic of fast muscles. The time course for development of the FDL and soleus is different, based on sarcomere structure with the soleus muscle developing clearly defined sarcomere structure earlier in gestation than the FDL. At 70 days gestation the FDL muscle had no clearly defined sarcomeres. Force (N cm-2) increased almost linearly between 70 and 140 days gestation in both muscle types and there was no difference between the Ca(2+)- and Sr(2+)-activated force throughout development. PMID- 10471990 TI - Shortening properties of two biochemically defined muscle fibre types of the Norway lobster Nephrops norvegicus L. AB - Mechanical properties of myofibrillar bundles from single chemically skinned fibres from the superficial abdominal flexor muscle of the Norway lobster Nephrops norvegicus were measured, and the protein content of these fibres was analysed by SDS-PAGE. Two slow fibre phenotypes (S1, S2) were distinguished on the basis of their myofibrillar protein assemblages. Data from 9 S1 and 8 S2 fibres obtained at similar sarcomere length demonstrate significant differences between the fibre types in maximal tension (N cm-2, S1: 10.5 +/- 3.9; S2: 3.1 +/- 0.8), in the delay of the peak of stretch activation (ms, S1: 122 +/- 18; S2: 412 +/- 202), in fibre stiffness (N cm-2 per nm half sarcomere, S1: 0.36 +/- 0.19; S2: 0.09 +/- 0.03) and in maximal shortening velocity (fibre length s-1, S1: 0.53 +/- 0.10; S2: 0.27 +/- 0.06). Furthermore, the maximal power output of the type S1 fibres was about five times larger than that of S2 fibres. The power output was maximal at lower loads in S1 fibres (relative load = 0.37 +/- 0.04) than in S2 fibres (relative load = 0.44 +/- 0.05). This study represents a comprehensive investigation of two slow muscle fibre types which are thought to be specialized for slow movements (S1 fibres) and for the postural control of the abdomen (S2 fibres). PMID- 10471991 TI - Differential response of the membrane systems involved in excitation-contraction coupling to early and later postnatal denervation in rat skeletal muscle. AB - We compared the morphological features of the membrane systems involved in excitation-contraction (E-C) coupling during early postnatal development stages in rat skeletal muscles (tibialis anterior) denervated either at birth or 7 days after birth. Four obvious structural changes are observed in the arrangement of the transverse (T) tubule network and the disposition of triads following early postnatal denervation: (1) an increase in the longitudinal segments of the T tubule network, (2) changes in the direction and disposition of triads, (3) the appearance of caveolae clusters, (4) the appearance of pentads and heptads (i.e. a close apposition of two or three T tubule elements with three or four elements of terminal cisternae of sarcoplasmic reticulum). The increased presence of longitudinal T tubules parallels the loss of cross striations, and this in turn is due to misalignment of the myofibrils. The clusters of caveolae appear almost exclusively in muscle fibres denervated at birth, and pentads and heptads are more frequently observed in muscles denervated at 7 days. The differential growth of muscle fibres in response to denervation leads to the formation of abnormal membrane systems involved in the E-C coupling with very unique morphological features, which differ from the case of denervation in adult muscle fibres. PMID- 10471992 TI - Sarcomeric binding pattern of exogenously added intact caldesmon and its C terminal 20-kDa fragment in skinned fibers of skeletal muscle. AB - Intact caldesmon and particularly the actin-binding C-terminal fragment (20-kDa) of caldesmon have been shown in skeletal muscle fibers to selectively displace low affinity, weakly bound cross-bridges from actin without significantly altering the actin attachment of force producing, strong binding cross-bridges (Brenner et al., 1991; Kraft et al., 1995a). However, the sarcomeric distribution and the specific binding of externally added caldesmon to the myofilaments of skeletal muscle fibers was not known. It was e.g., unclear whether caldesmon binds along actin in a manner similar to tropomyosin or whether it also binds to myosin. In this study, we determined the binding pattern of exogenously added intact caldesmon and its C-terminal 20-kDa fragment, respectively, in MgATP relaxed rabbit skeletal muscle fibers using electron (EM) and confocal fluorescence microscopy (CFM). EM showed that similar to what has been demonstrated earlier for smooth muscle thin filaments (Lehman et al., 1989), intact caldesmon binds periodically every 38 nm along the thin filaments. CFM revealed that rhodamine-labeled intact caldesmon and the 20-kDa caldesmon fragment bind along nearly the entire length of the thin filaments. A portion of the I-band near the Z-line appears unlabeled, both when equilibrated at normal and long sarcomere lengths. The width of the unlabeled region seems to depend on ionic strength. The 20-kDa C-terminal caldesmon fragment binds in essentially the same pattern as intact caldesmon. This indicates that the high fluorescence intensity in the overlap region seen with intact caldesmon does not depend on caldesmon binding to myosin. X-ray diffraction was used to monitor the effects of filament lattice. Intact caldesmon at > 0.3 mg/ml induced disorder in the myofilament lattice. No such disordering was observed, however, when fibers were equilibrated with up to 0.8 mg/ml of the 20-kDa caldesmon fragment. PMID- 10471995 TI - The Annual Meeting on Muscle Contraction and Cell Motility. Nagoya, Japan, 6-8 January 1999. PMID- 10471993 TI - Utrophin and dystrophin-associated glycoproteins in normal and dystrophin deficient cardiac muscle. AB - In this study, various members of the dystrophin family (dystrophin, the short dystrophin product Dp 71, utrophin and DRP2), and different members of the dystrophin-associated glycoprotein (DAG) complex (beta-dystroglycan, alpha-, beta , gamma- and delta-sarcoglycans) were localized in bovine cardiac muscle using a battery of specific antibodies. We have established that dystrophin is exclusively associated with beta-dystroglycan and both alpha- and delta sarcoglycans in cardiac muscle cell membranes. In contrast, utrophin is a specific component of intercalated disks together with beta- and gamma sarcoglycans, while beta-dystroglycan, alpha- and delta-sarcoglycans are not present. Dp 71 is mainly localized at the T tubule transverse area. In dystrophin deficient cardiac muscle, utrophin and beta-sarcoglycan were observed in intercalated disks and at the sarcolemma of each cardiocyte. Our results revealed that complexes of associated glycoproteins differ in cardiac muscle when associated with dystrophin or utrophin. Despite the described sequence homologies between dystrophin and utrophin, the present results indicate that these proteins have different roles in some specific cardiac cell areas. PMID- 10471996 TI - Clavicle malunion. AB - The clavicle fracture that has united with deformity or shortening may have an adverse effect on normal shoulder girdle function. We report on 4 patients in whom a malunited fracture of the clavicle was believed to be a contributing factor to shoulder girdle dysfunction. In each patient, the functional status of the involved limb was improved after corrective osteotomy at the site of deformity, realignment, and plate fixation. PMID- 10471994 TI - Characterisation of troponin-T from salmonid fish. AB - Five major troponin-T isoforms were isolated from the myotomal muscles of Atlantic salmon: three from fast muscle (Tn-T1F, Tn-T2F and Tn-T3F) and two from slow muscle (Tn-T1S and Tn-T2S). In addition to their presence in troponin preparations, these proteins were also recognised to be Tn-T on the basis of immunoreaction with anti-troponin-T antibodies and partial amino acid sequence. The electrophoretic mobility in the presence of SDS of the various Tn-Ts increases in the order: 1S < 1F < 2S < 2F < or = 3F. Compositional analysis shows that the higher M(r) forms (1F and 1S) contain considerably more proline, glutamic acid and alanine than the lower-M(r) forms (2F, 3F and 2S). Every isoform lacks cysteine and phosphoserine is present only in isoforms 2F and 3F. All of the Tn-Ts, with the exception of isoform 1F, are N-terminally blocked. CNBr fragments from same cell type Tn-Ts yield identical sequences over at least fifteen Edman cycles. Two full-length cDNA sequences, presumed to represent 1S and 3F, or isoforms that are highly similar, are reported. As documented for higher vertebrate Tn-Ts, the predicted primary structures display a non-uniform distribution of charged amino acids and greater divergence at each end than in the central section. The most striking difference between the two salmonid proteins is the presence of a N-terminal (proline-, glutamic acid- and alanine rich) extension of about fifty amino acids in Tn-T1s (278 amino acids) that is missing from the fast muscle Tn-T (223 amino acids). The sequences also differ in that 1S lacks the known phosphorylation site while the fast-type isoform contains serine next to the initiating methionine. Of the two, the slow isoform has accumulated the greater number of substitutions. PMID- 10471997 TI - Surgery for rheumatoid arthritis of the elbow: a comparison of radial-head excision and synovectomy with total elbow replacement. AB - The results of total elbow replacement (TER) in 45 elbows of 38 patients with rheumatoid arthritis were compared with results of radial head excision with synovectomy (RHES) in 45 age-matched patients treated in the same unit. The groups were similar with respect to duration of disease and preoperative clinical status, although pain was of longer duration and slightly more severe in the TER group. Failure was defined as the onset of moderate or severe pain after surgery or revision surgery for any reason. Reduction in pain was greater after TER than after RHES (P < .05). Recurrence of pain was common after RHES but was not seen after TER. Movement increased by a similar amount in each group. Complications were more frequent and more serious after TER (4 dislocations, 4 ulnar nerve dysfunctions, 1 significant wound breakdown) than after RHES (2 ulnar nerve dysfunctions, 1 transient wound discharge). Complications after TER were most common in patients who had previous RHES. On survival analysis, TER results were better than RHES results in each successive year. Cumulative survival rates at 10 years were 85% for TER and 69% for RHES, but the difference in rates was not statistically significant. In the medium term, TER relieves pain more reliably than RHES and its use is justified despite the greater risk of complications. In view of the paucity of long-term results for TER, RHES may retain a role in younger patients or in those whose symptoms are related mainly to the radiohumeral joint. PMID- 10471998 TI - Age-related prevalence of rotator cuff tears in asymptomatic shoulders. AB - To determine the prevalence of rotator cuff tears in asymptomatic shoulders we conducted a prospective clinical and ultrasonographic study of 411 volunteers. We anticipated an age-dependent outcome and divided the patients into 4 age-groups. Overall, we found evidence of a rotator cuff tear in 23% of the patients. In group 1 (aged 50 to 59 years), 13% (22 of 167) of the patients had tears; in group 2 (aged 60 to 69 years), 20% (22 of 108) of the patients had tears; in group 3 (aged 70 to 79 years), 31% (27 of 87) of the patients had tears; and in group 4 (age > 80 years), 51% (25 of 49) of the patients had tears. An astonishingly high rate of rotator cuff tears in patients with asymptomatic shoulders was thus demonstrated with increasing patient age. At this stage it remains unclear, however, which parameters convert an asymptomatic rotator cuff tear into a symptomatic tear. As a result, rotator cuff tears must to a certain extent be regarded as "normal" degenerative attrition, not necessarily causing pain and functional impairment. PMID- 10471999 TI - Restraining patients and shoulder dislocations during seizures. AB - We describe 3 patients whose shoulders dislocated as the movements of the arm were restricted during a generalized tonic clonic seizure over an 18-month period. The first patient had both shoulders dislocated when observers sat on his arms during the convulsion. The second patient had a convulsion while in a forced lateral decubitus position and dislocated the shoulder on that side. The third patient dislocated the shoulder and fractured the acromion as she was held by her arms in a chair during a convulsion. Despite the large number of patients with refractory epilepsy under our care, no cases of spontaneous shoulder dislocation occurred during that period of time. PMID- 10472000 TI - Relation between the painful shoulder and the cervical spine with narrow canal in patients without obvious radiculopathy. AB - It is well known that cervical radiculopathy sometimes causes shoulder pain. Hypothesizing that the cause of painful shoulder is related to the cervical spine in the absence of obvious radiculopathy, we measured the anteroposterior diameter of the spinal canal and the range of motion of the cervical spine in patients with painful shoulder on lateral cervical radiographs of the spine. Painful shoulder was diagnosed in 76 patients (24 men and 52 women; mean age 57.6 years). Patients who reported neck pain or numbness of the upper limbs and patients with neurologic abnormalities were excluded from this study. A control group of 54 asymptomatic volunteers (27 men and 27 women; mean age 55.5 years) was formed. The difference in age between the patient group and the control group was not significant. The anteroposterior diameter of the spinal canal at C5 and C6 in the painful-shoulder group (C5: 12.74 mm; C6: 12.76 mm) was significantly narrower than in the control group (C5: 13.60 mm; C6: 13.79 mm). The range of motion was greatest at C4-5 and smallest at C2-3 in both groups; and there was no significant difference in the range of motion between the painful-shoulder group and the control group. When women only or men only were assessed, the results were nearly the same between groups. The cervical spine without obvious radiculopathy appears to be involved in patients with a painful shoulder. We speculate that the shoulder is affected by irritation of a cervical nerve root or referred pain. PMID- 10472001 TI - Surgical anatomy of the posterior shoulder: effects of arm position and anterior inferior capsular shift. AB - The purposes of this study were to evaluate anatomically various surgical intervals to the posterior shoulder and to determine the effects of varying arm positions and anterior-inferior capsular shift (AICS) on the relation of the posterior neurovascular structures to fixed bony landmarks. Fourteen cadaveric shoulders were dissected. The posterior surgical anatomy was defined, and the distances from fixed bony landmarks to neurovascular and musculotendinous structures were determined with digital calipers. Measurements were made with the arm in various positions and repeated after AICS. The most direct anatomic approach to the posterior shoulder was through a deltoid split in the raphe from the posterolateral corner of the acromion (PLCA), followed by an infraspinatus (IS) splitting incision. The IS/teres minor interval was at the inferior aspect of the glenoid rim and was difficult to locate in all specimens. The distance to the axillary nerve from the PLCA averaged 65 mm and decreased by an average of 14 mm (22%) with abduction and by 19 mm (29%) with extension. The posterior humeral circumflex artery was located along the humeral neck and was vulnerable to injury during lateral capsular dissection. The suprascapular nerve had multiple branches to the IS with most penetrating the muscle at its inferior portion. The closest branch to the glenoid rim was an average of 20 mm medial from it. No branch entered at the level of the IS raphe. The anatomic relations of the suprascapular nerve were unchanged after AICS. On the basis of this study, surgical exposure of the posterior shoulder with a deltoid split from the PLCA, followed by an IS split, appears to be anatomically safe. The arm position should be in neutral rotation, especially if previous anterior capsular procedures have been performed, which can alter the posterior neurovascular anatomic relations. PMID- 10472002 TI - Troubleshooting the supraspinatus outlet view. AB - Radiographic evaluation of the acromion is integral to the management of subacromial impingement syndrome. The supraspinatus outlet view reveals acromial morphologic traits, acromial slope, subacromial excrescences, and inferior osteophytes at the acromioclavicular joint. The major disadvantage of the outlet view is reproducibility. The purpose of this article is to describe technical factors that allow for obtaining this view consistently. In this study 7 different outlet views were obtained of the shoulder of an asmptomatic, voluntary subject. One view was obtained correctly; the other 6 were taken by simulating common technical errors. Each radiograph was then evaluated to determine how each technical error affected the radiographic image. Guidelines were then formulated for obtaining a proper outlet view on no more than 2 attempts. Correction of a faulty supraspinatus outlet view should be guided by the shape of the scapular body, the position of the floor of the supraspinatus fossa, and the distance between the humeral head and acromion. PMID- 10472003 TI - Translation of the glenohumeral joint in patients with anterior instability: awake examination versus examination with the patient under anesthesia. AB - Fifty patients with a clinical diagnosis of traumatic anterior shoulder instability underwent bilateral shoulder translation testing while both awake and under anesthesia. Each patient was examined by 2 surgeons following guidelines developed by the American Shoulder and Elbow Surgeons. A single translation grade was established for anterior, posterior, and inferior directions. A comparison of means was performed with a paired t test. The mean anterior translation grade was significantly higher on the affected side when compared with that of the unaffected side both during awake examination and during examination with the patient under anesthesia (EUA). Ipsilateral comparison revealed significantly greater translation for both affected and unaffected shoulders in anterior, posterior, and inferior directions during EUA than during awake examination. Side to-side comparison of posterior and inferior translation was similar for both awake examination and EUA. Clinical translation testing was helpful in the diagnosis of anterior shoulder instability. Side-to-side differences were subtle while awake and more apparent during EUA. The usefulness of awake translation testing for traumatic anterior instability was not clearly demonstrated; however, EUA provides helpful information to confirm the direction and degree of instability. PMID- 10472004 TI - Contribution of the passive properties of the rotator cuff to glenohumeral stability during anterior-posterior loading. AB - The passive properties of the rotator cuff have been shown to provide some stability during anterior-posterior (AP) translation. However, the relative importance of the rotator cuff to joint stability remains unclear. The purpose of this study was to quantify the force contributions of the rotator cuff and of capsuloligamentous structures at the glenohumeral joint during AP loading. We hypothesized that the rotator cuff acts as a significant passive stabilizer of the glenohumeral joint and that its contribution to joint stability is comparable to the contribution made by the components of the glenohumeral capsule. A robotic/universal force-moment sensor testing system was used to determine both the multiple "degrees of freedom" joint motion and the in situ force carried by each soft tissue structure during application of an 89N AP load at 4 abduction angles. The percent contribution of the rotator cuff to the resisting force of the intact joint during AP loading was significantly greater during posterior loading (35% +/- 26%) than during anterior loading at 60 degrees of abduction (P < .05). The contribution of the rotator cuff (i.e., 29% +/- 16% at 30 degrees of abduction) was found to be significantly greater than the contributions of the capsule components during posterior loading at 30 degrees, 60 degrees, and 90 degrees of abduction (P < .05). However, no differences could be found between the respective contributions of the rotator cuff and the capsule components during anterior loading. The results support our hypothesis and suggest that passive tension in the rotator cuff plays a more significant role than other soft tissue structures in resisting posterior loads at the glenohumeral joint. The important role of the rotator cuff during posterior loading may be a result of the thin posterior joint capsule compared with the anterior capsule, which has several thickenings. This information increases our understanding of posterior stability at the glenohumeral joint during clinical laxity tests. PMID- 10472005 TI - Self-assessment questionnaires document substantial variability in the clinical expression of rotator cuff tears. AB - The goal of this investigation was to document the variability in the clinical expression of full-thickness rotator cuff tears with practical and standardized patient self-assessment tools. One-hundred twenty-three consecutive patients with full-thickness cuff tears diagnosed by standard cuff-imaging methods (sonography, arthrography, or magnetic resonance imagery) assessed their own shoulder function and health status with the Simple Shoulder Test and the Short Form 36, respectively. As a group, these patients were substantially compromised in their ability to perform the functions of the Simple Shoulder Test and in the Short Form 36 scales of physical role, physical function, and comfort. As individuals, however, their self-assessments varied widely. The standard deviations were often greater than 50% of the mean and the range of responses often covered the entire scale from the minimum possible score to the maximum possible score. These results show the importance of documenting the clinical expression of cuff tears in patients at initial evaluation and when treatment is being considered. The results also show the practicality of standardized self-assessment questionnaires in such documentation. PMID- 10472006 TI - The teres major muscle: an anatomic study of its use as a tendon transfer. AB - Eleven fresh-frozen cadaver shoulders were dissected to define the anatomy of the teres major muscle and tendon and to determine the muscle's potential for use as a tendon transfer to the humeral head. Of the 11 specimens, 7 had Mathes type II circulation. The primary and secondary pedicles, from the circumflex scapular artery, entered the muscle 4.1 cm and 0.5 cm from the scapula, respectively. The lower subscapular nerve entered 4.1 cm from the scapula. Mean tendon and muscle lengths were 2.0 and 11.8 cm, respectively. As a unipolar transfer, the tendon reached the greater tuberosity in all but 1 specimen. The bipolar transfer offered numerous theoretical possibilities. We believe that the teres major has an appropriate vascular supply and adequate length to make it suitable for tendon transfer to the humeral head. PMID- 10472007 TI - Thermal modification of collagen. AB - Shoulder capsular shrinkage has recently been proposed as a therapeutic modality in a select group of patients with instability. Basic science research studying the mechanism of collagen shrinkage and the effect of shrinkage on the tissue's mechanical properties is essential to define the ideal process by which to achieve optimal tissue shrinkage. Tissue shrinkage is a function of both time and temperature. This relationship was studied, and a model was derived to describe the relationship mathematically. Tissue shrinkage rate was extremely sensitive to temperature changes. The purpose of this study, was to shrink collagenous tissue thermally and then to measure the mechanical property changes as a function of tissue shrinkage. Uniaxial tensile testing of normal and heat-shrunken bovine tendon was carried out, and a model was developed to express the relationship between shrinkage and mechanical properties. We found that the mechanical properties decreased with increasing shrinkage, and that the maximal allowable shrinkage before significant material property changes occurred was between 15% to 20%. Ultrastructural analysis with transmission electron microscopy showed denaturation of the collagen fibrillar structure and provided direct support for the observed material changes. PMID- 10472008 TI - Arthroscopic treatment of multidirectional instability. AB - Multidirectional instability of the shoulder, described by Neer and Foster, has been treated surgically with the inferior capsular shift procedure. The small number of reports on mid-term outcomes indicate that good to excellent results have been obtained in 75% to 100% of cases. Arthroscopic treatment of multidirectional instability has been previously described. The purpose of this study was to review the results of the arthroscopic capsular shift procedure with a minimum follow-up of 2 years. A retrospective study was performed on 25 patients who underwent an arthroscopic capsular shift performed with the transglenoid technique between January 1990 and December 1993. All patients had earlier not responded to an extensive course of physical therapy. Excluded from the study were patients who had undergone a previous arthroscopic capsular shift or any other procedure, arthroscopic or open, for the shoulder. Average patient age was 26.4 years. There were 20 male and 5 female patients. Sixteen of the affected shoulders involved the dominant extremity. All patients had a history of asymptomatic subluxation that slowly progressed to symptomatic subluxation. Eleven patients had a history of dislocation. Thirteen patients were athletes who were symptomatic in their chosen sport, whereas the other patients were symptomatic in activities of daily living. All patients were examined while they were under anesthesia and had positive results on the sulcus test in abduction with associated anterior instability, posterior instability, or both. Follow-up evaluation was performed with patient interview and examination. All 25 patients were available for follow-up, which occurred an average of 60 months (range 36 to 80 months) after operation. Three patients had episodes of instability after the operation. The average Bankart score was 95 (range of 50 to 100). All but 1 patient had regained full symmetric range of motion by follow-up. Twenty-one (88%) patients had a satisfactory result according to the Neer system. Results of treatment with the arthroscopic capsular shift procedure for multidirectional instability of the shoulder appear to be comparable to those of the open inferior capsular shift. PMID- 10472009 TI - A standardized method for assessment of elbow function. Research Committee, American Shoulder and Elbow Surgeons. AB - The American Shoulder and Elbow Surgeons have adopted a standardized form for assessment of the elbow. This form was developed by the Research Committee of the American Shoulder and Elbow Surgeons and subsequently adopted by the membership. The patient self-evaluation section contains visual analog scales for pain and a series of questions relating to function of the extremity. The responses to the questions are scored on a 4-point ordinal scale. The physician assessment section has 4 parts: motion, stability, strength, and physical findings. It is hoped that adoption of this method of data collection will stimulate multicenter studies and improve communication between professionals who assess and treat patients with elbow disorders. PMID- 10472011 TI - Adverse effects of extensive clavicular resections and a suggested method of reconstruction. PMID- 10472010 TI - Radial head fractures associated with elbow dislocations treated by immediate stabilization and early motion. AB - Twenty-one elbow dislocations with an associated radial head fracture were treated with immediate joint reduction, stabilization, and early range-of-motion exercises. In all cases initial treatment involved closed reduction of the ulnohumeral joint. For those cases involving minimally displaced and a few moderately displaced radial head fractures, treatment consisted of benign neglect (4 of 21). Of the more severely displaced fractures (17 of 21), 9 were treated with open-reduction internal fixation and 8 with immediate silicone head replacement. Despite radial head treatment, 6 of these cases remained unstable, prompting primary repair of collateral ligaments; 3 eventually required application of a hinged fixator as a salvage option. Results confirmed that initial radial head displacement predicts functional outcome. Our study demonstrates that fracture dislocations of the elbow demand a broad consideration of treatment options and that reconstruction of elbow stability requires either primary repair of collateral ligaments or the possible use of a hinged fixator device. PMID- 10472012 TI - Enormous glomus tumor of the shoulder. PMID- 10472013 TI - Total elbow arthroplasty for treatment of elbow arthritis. PMID- 10472014 TI - General vs specialist medicine. PMID- 10472015 TI - The National Institute for Clinical Excellence: the government's agenda and the College's role. PMID- 10472016 TI - New drug treatments in psychiatric disease. PMID- 10472017 TI - Prion related disorders. PMID- 10472018 TI - Multiple sclerosis and its treatment. AB - Multiple sclerosis (MS) is a common neurological disorder responsible for substantial neurological morbidity. Although it is considered to be an autoimmune demyelinating disease of the central nervous system (CNS), mediated by antigen specific CD4+ T helper (Th1) T-cells, therapeutic strategies aimed at generalised immunosuppression have been disappointing. Recently, immunomodulatory therapies like interferon (IFN)-beta and glatiramer acetate have proved more effective. They reduce the rate and severity of clinical relapses and, in the case of IFN beta, delay the rate of disease progression. Symptomatic therapies and rehabilitation, however, remain the mainstay of treatment for the majority of patients with MS. The immunopathogenesis of MS and its treatments, both disease modifying and symptomatic, are reviewed below. PMID- 10472019 TI - Disorders of the basal ganglia and their modern management. PMID- 10472020 TI - The use of new antiepileptic drugs. PMID- 10472021 TI - Primary care groups and primary care trusts. PMID- 10472022 TI - General internal medicine and specialty medicine--time to rethink the relationship. PMID- 10472023 TI - Audit of resuscitation decisions has little impact on clinical practice. AB - OBJECTIVE: To determine if clinical audit leads to sustained improvement in resuscitation decision-making. METHOD: Analysis of data from five studies performed over nine years between 1989 and 1998. Two of the surveys included all medical and elderly patients, while three surveys in 1993, 1994 and 1996 included only patients over 75 years of age. RESULTS: The three surveys involving only elderly patients revealed significant improvement in recording the decisions made on resuscitation (73% vs 50%; p = 0.02) over a two-month audit cycle, but this improvement was not maintained. Clinical audit and the use of a proforma improved neither the practice of discussing resuscitation with patients nor the involvement of consultants in the decision-making process over the nine-year period. CONCLUSION: Clinical audit may lead to short-term improvements in resuscitation decision-making, but this improvement does not appear to be sustained over time. Other measures need to be considered to improve practice in this area. PMID- 10472024 TI - Measuring the prevalence of disease in middle-aged British men. AB - OBJECTIVE: To identify and validate a measure of disease prevalence in a town or other subpopulation for the purpose of estimating variations in morbidity and health care needs between groups. DESIGN: A national prospective study of cardiovascular disease based in 24 British towns. SUBJECTS: 7,735 men aged 40-59 years sampled from 24 general practice age-sex registers between 1978 and 1980. METHOD: A questionnaire administered at recruitment and a postal questionnaire five years later (98% response) asked if the subject recalled ever having had a diagnosis by a doctor of any of 12 listed conditions. In addition, the men were asked about regular medication, occupation, lifestyle and perceived health status. All-cause mortality data were collected during 13.5 years of follow-up. RESULTS: Prevalence of recalled doctor diagnoses varied between the towns; for cardiovascular disease, it correlated significantly with other health indices and personal characteristics and with standardised mortality ratios for the town. Consistency of reporting over time and mortality outcome after 13.5 years also supported the validity of patient recall. CONCLUSIONS: A simple standardised questionnaire using patient recall of common conditions diagnosed by a doctor provides a plausible, valid measure of variations in disease burden between communities. PMID- 10472025 TI - Training in academic medicine: a way forward for the new millennium. A discussion document from the Academic Medicine Committee of the Royal College of Physicians. AB - Three schemes are presented for discussion whereby physicians undergoing postdoctoral training can combine a period of research training with their clinical training and so enable those who wish to follow a career in academic medicine to do so, or alternatively to revert to a clinical career. The training arrangements for those wishing to take up clinical academic medicine have hitherto been uncertain and hence unattractive to some. As well as encouraging more high-calibre trainees into academic medicine, the training programmes described are intended to bring greater clarity to those responsible for academic and clinical training and to those who fund research. PMID- 10472026 TI - Ethical decisions at the end of life. AB - A conference organised by Dr Gillian Ford, on behalf of the College Ethical and Palliative Medicine Committees, was held at the Royal College of Physicians of London on 24 March 1999. It attracted over 200 delegates, two-thirds of whom were doctors. Issues covered included hydration and dehydration in the dying patient, talking to patients and their families, the management of futility, caring for patients at home, listening to children, the limits of autonomy and a debate on the principle of 'double effect'. PMID- 10472027 TI - The poisons and narcotics of the Amazonian Indians. AB - The indigenous tribes of the Amazonian rainforest have discovered a vast array of poisons and narcotics from the plant species which surround them. Examples are given from the author's personal experience of a variety of arrow poisons, fish poisons and narcotics used by six different tribes. There is a great variety of different plant species used for each category and no two tribes studied employ exactly the same array of plant species. This makes it important to survey a wide range of tribes and to document this information before it is too late as these people are fast losing their traditional culture. Several of the substances used by Amazonian Indians, such as curare and coca, have entered into western medicine and others such as rotenone, an ingredient of their fish poison, has become a useful insecticide. Further useful chemicals are likely to be discovered from ethnobotanical work among indigenous peoples and it is vital that any commercial gain from their knowledge be of direct benefit to these people in accordance with the rules of the Convention on Biological Diversity. PMID- 10472028 TI - The value and quality of life. AB - A medical view on 'the value of life' can be inferred from medical accounts of the quality of life: a life has value if it embodies certain qualities. Scales have been developed to quantify quality of life. While the term 'quality of life', is used frequently in everyday discourse, perceptions of what it might actually mean differ greatly and are often incompatible. This incompatibility can be illustrated through an examination and development of the Greek myth of Sisyphus. The different models that the author explores rest on 'significant toil', 'choice', 'happiness or well-being', or 'social factors' being the prerequisite for quality of existence. These models are incommensurable and, as intangible concepts, cannot be quantified. Decision-making in medicine does not require a complex evaluation of quality of life: it consists of the doctor's offer of treatment based on the best evidence, and the patient's consent to, or refusal of, that offer. Apart from the need to obtain consent, the main ethical constraint on the doctor is equity. PMID- 10472029 TI - Villain and victim: the kidney and high blood pressure in the nineteenth century. PMID- 10472030 TI - Me and my illness: migraineuse. PMID- 10472031 TI - Thrombolysis nurse. PMID- 10472032 TI - Thrombolysis in inferior myocardial infarct--unhelpful? PMID- 10472033 TI - A training course for the UEB examination. PMID- 10472034 TI - Cyberclinic in rheumatology. PMID- 10472035 TI - SHO experience in general practice. PMID- 10472036 TI - An overview of the clinical use of dynamic posturography in the differential diagnosis of balance disorders. AB - Dynamic posturography comprises a series of balance control tests which help physicians overcome numerous diagnostic and treatment challenges arising when examining patients complaining of a debilitating balance disorder. These challenges include the specific differential diagnosis, documentation of symptoms and assessment of functional disability. It must be determined whether the cause of the disability is an organic sensory, deficit, a central nervous system (CNS) lesion or a non-organic (that is, possibly psychogenic or just overtly simulated) disorder. This review is targeted towards providing the reader (a) an overview of the effects sensorimotor deficits have on balance control, specifically vestibulospinal and proprioceptive reflex deficits; and, (b) how these effects may be assessed objectively in a clinical setting to differentiate between various organic and non-organic balance-disorders. The techniques used to study these effects are based on the analysis of both rapid balance-correcting and slow balance-stabilizing responses to fast and slow movements in the pitch plane of the support surface on which the test subject stands. PMID- 10472037 TI - The activities-specific balance confidence scale and the dizziness handicap inventory: a comparison. AB - Vestibular dysfunction can have a tremendous impact on an individual's quality of life. The purpose of this paper is to determine if the level of handicap reported by individuals on the Dizziness Handicap Inventory (DHI), an inventory developed for use with individuals with complaints of dizziness symptoms, will be consistent with that reported on the Activities-specific Balance Confidence Scale (ABC), a tool developed for use with elderly individuals that attempts to assess a person's confidence level in performing activities of daily living (ADL's). A sample of convenience was used consisting of 71 subjects (15 males and 56 females) from a local Balance and Vestibular Clinic. The subjects ranged in age from 26 to 88 years of age. Both the DHI and the ABC were administered as part of an initial physical therapy evaluation to new patients at the clinic. A moderately strong negative correlation was found between the scores of the two inventories (Ts = -0.6350). The results suggest that the ABC is a valid tool for use with individuals with complaints of dizziness. PMID- 10472038 TI - Further evidence for age-related deficits in human postural function. AB - The aim of the present study was to investigate the effects of aging on postural function in human subjects. Subjects aged 18-39 years (n = 24), 40-59 years (n = 24) and 60 years or older (n = 27) were tested for postural stability using an electronic balance platform. There was a significant increase in postural instability with increasing age (P < 0.0001), and as the conditions of testing became more difficult (P < 0.0001) (e.g., with eyes closed). Elderly males were found to have significantly poorer postural control than elderly females in the visual-vestibular conflict conditions (P < 0.05). These results further demonstrate a decline in postural function with increasing age, which may be related to deterioration in the peripheral or central vestibular systems and may be partly responsible for the high incidence of falls in the elderly. PMID- 10472039 TI - Voluntary movement strategies of individuals with unilateral peripheral vestibular hypofunction. AB - This study compared voluntary movement strategies of patients with unilateral peripheral vestibular hypofunction with those of age-matched healthy control subjects. All subjects performed three voluntary movement tasks with their dominant upper extremity: a forward flexion arm movement through 90 degrees, a reach to an overhead target, and a reach to a side target. Subjects performed the movement tasks sitting and standing (Body Position), and under precued and choice reaction time (RT) conditions (Task Certainty). Measures of motor planning and movement execution included RT and movement time (MT), respectively. Statistical analysis included separate Group x Task Certainty x Body Position ANOVA calculations for each task. Across tasks, results suggested no between group differences for RT. A Task Certainty main effect for the side and overhead tasks indicated that the choice RT situation resulted in longer RTs as compared to the precued RT condition. Movement time differed between the two groups. Across all three voluntary movement tasks, vestibular impaired subjects moved more slowly than control subjects. Providing vestibular subjects with a precue did not bring MT performance to the level of controls. Body position influenced MT for the side task only. Across both groups of subjects, MT for the side task was longer when performed in the standing position. The results of this study suggest that individuals with unilateral peripheral vestibular hypofunction initiate voluntary movement responses with similar timing as control subjects, but require more time to complete the movement. Vestibular rehabilitation should include goal-directed movement and should address issues of movement speed. PMID- 10472040 TI - Surface height effects on postural control: a hypothesis for a stiffness strategy for stance. AB - One possible factor influencing the control of upright stance is the perceived threat to one's personal safety, i.e. balance confidence. We explored this factor by examining the control of stationary stance when standing on an elevated platform under various conditions of reduced visual and vestibular inputs. Twenty eight adults (14 male and 14 female, mean age = 23.5 years) participated in the experiment. Postural control was examined by recording the amplitude variability (RMS) and mean power frequency (MPF) of center of pressure excursions (COP) over a 2-minute interval while participants stood in a normal stance on a low (0.19 m) and a high (0.81 m) platform with toes positioned either at or away from the edge of the platform. Vision was manipulated through eyes open and eyes closed trials. Vestibular input was reduced by tilting the head into extension [1]. Anterior posterior RMS and MPF of COP were significantly influenced by an interaction between surface height and vision. When vision was available, a significant decrease in RMS was observed during quiet standing on a high surface compared to a low surface independent of step restriction. When vision was available MPF increased when subjects were raised from a low to a high surface. The mean position of the COP was significantly influenced by an interaction between height and step restriction. Differences in RMS and MPF responses to height manipulation were observed between genders in eyes closed conditions. Vestibular input influenced postural control at both low and high levels with significant increases in RMS when vestibular input was reduced. The reciprocal changes observed in RMS and MPF suggest modifications to postural control through changes in ankle stiffness. Vision appears to play a role in increasing ankle stiffness when balance confidence is compromised. PMID- 10472041 TI - Circular vection in patients with age-related macular degeneration. AB - We have used optokinetic stimulation in patients with unilateral age-related macular degeneration (AMD) and central scotoma to investigate the possible contribution of the central visual field to circular vection (CV). Six patients aged 42-73 years with unilateral AMD and an aged-matched control group of nine elderly adults aged 47-75 years were examined. Monocular visual field defects were verified with the Goldmann perimeter by kinetic perimetry. The device used to induce CV was a random dot pattern projected onto a hemispherical dome with a radius of 75 cm. The pattern was rotated horizontally at a constant acceleration of 1 deg/s2. Monocular stimuli were randomly repeated two to three times in both temporal-nasal (T-N) and nasal-temporal (N-T) directions. The latency of onset of CV was measured for each stimulus presentation. In the age-matched control group the CV latencies varied from 4.2 to 72.0 s. In each case, however, the CV latencies were stable. No significant differences in CV latencies were found between right and left eyes in both stimulus directions (p > 0.05). In patients with AMD, no statistically significant difference in CV latency was found between the affected and unaffected eyes (p > 0.05). Marked central visual field loss in AMD does not significantly impair peripherally induced CV. Our results are compatible with the hypothesis that the peripheral retina dominates CV. PMID- 10472043 TI - Introduction: gramicidin, a model ion channel. PMID- 10472042 TI - Model-based study of the human cupular time constant. AB - The time constant of the angular vestibulo-ocular reflex (aVOR), measured from the response to steps of rotation about a yaw axis, has frequently been estimated as a single exponential. However, the slow phase velocity envelope during per- or post-rotatory nystagmus is more accurately represented by two exponential modes. One represents activity in the vestibular nerve induced by deflection of the cupula, the other by activation that the input from the canals produces in the central velocity storage integrator. The sum of the cupula and the integrator responses describes the overall response of slow phase eye velocity and can be approximated by a double exponential. Frequently, there is a plateau in the initial portion of eye velocity response, but this may be masked by habituation, making the cupula contribution unobservable and impossible to estimate. Using a model-based technique to analyze responses with a clear plateau, we estimated peripheral and central vestibular time constants by double exponential fits to slow phase eye velocity. Cupular time constants were varied from 1 to 10 s to identify values that gave optimal fits of the data according to a Chi-square criterion. The mean cupular time constant for 10 human subjects was 4.2 +/- 0.6 s. Fits of the data were also good for time constants between 3.5 to 7 s, but not for 1 to 3 or 7.5 to 10 s. The estimated cupular time constants also fit responses where there was no plateau. In 8 monkeys, cupular time constants were estimated as 3.9 +/- 0.5 s, which agreed with those derived from activity in the vestibular nerve. There was no difference between monkey and human cupular time constants from these estimates. It is likely that the human cupular time constant is similar to that of the monkey and shorter than previously thought. PMID- 10472044 TI - Correlations of structure, dynamics and function in the gramicidin channel by solid-state NMR spectroscopy. AB - The high resolution structure of the gramicidin A channel has been determined in a lamellar phase environment using solid-state NMR spectroscopy. While the fold is similar to previous characterizations, channel function is exquisitely dependent on structural detail. There is essentially no structural change upon cation binding and no significant change in dynamics. The cations appear to be adequately solvated in their binding site by no more than two carbonyls and no fewer than three water molecules at any one time. The relatively large number of water molecules allows for geometric flexibility and little selectivity among monovalent cations. However, the dehydration energies of cations clearly explain the selectivity for monovalent versus divalent cations. Moreover, the binding site is shown to be delocalized, resulting in a shallow potential energy well so that efficient cation conductance can be realized. The potential energy barrier at the bilayer centre has been shown to be rate limiting under certain circumstances through a correlation between conductance and the electrostatic interactions between cations at the gramicidin monomer-monomer junction and the indole dipole moments at the lipid-water interface. The dynamics are functionally important. The time-scale for carbonyl fluctuations about the C alpha-C alpha axis and kinetic rates for cation movement in the channel are the same, suggesting a correlation between molecular dynamics and kinetics. These functional correlations will be described in light of the recent K+ channel structure and the biological challenge to achieve both selectivity and efficiency. PMID- 10472045 TI - X-ray crystallographic structures of gramicidin and their relation to the Streptomyces lividans potassium channel structure. AB - Gramicidin has been used extensively as a model system for structure/function studies of ion channels. Long before crystals of other ion channel proteins were produced, crystals of gramicidin had been prepared, even though it was many years before the first forms of those crystals were solved. There now exist a large number of crystal structures of both uncomplexed and ion-complexed forms of gramicidin crystallized from organic solvents. In all these crystals, the molecules are double helices, although they differ in helical pitch, handedness and side chain orientations, depending on the conditions used for crystallization. Since many of these structures have been discussed in detail in a recent review (Wallace 1998), this chapter concentrates on recently reported structures and how they relate to previously described X-ray and NMR structures. It also discusses how the crystal structure of a K+ complex of gramicidin relates to the recently solved structure of a K+ complex of the potassium channel from Streptomyces lividans and argues that this demonstrates that gramicidin is indeed a good model structure for biological ion channels, despite the presence of D amino acids in its sequence. PMID- 10472046 TI - The effects of ion binding. PMID- 10472047 TI - Design and characterization of gramicidin channels with side chain or backbone mutations. AB - Mutations and chemical substitutions of amino acid side chains and backbone atoms have proved vital for understanding the folding, structure and function of gramicidin channels in phospholipid membranes. The channel's pore is lined by peptide backbone groups; their importance for channel structure and function is shown by a single amide-to-ester replacement within the backbone, which greatly reduces the resulting channel conductance and lifetime. The four tryptophans and the intervening leucines together govern the formation and dissociation of conducting channels from single-stranded subunits. Conducting double-stranded gramicidin conformations (channels) occur rarely in membranes--except when the sequence has been altered to permit special arrangements of tryptophans or (infrequently) in unusually thick membranes. The tryptophans anchor the single stranded channels to the membrane/solution interface, and the indole dipoles promote cation transport through the channels. Removal of any indole dipole reduces ion conductance; whereas 5-fluorination of an indole, which increases its dipole moment, enhances ion conductance. Some sequence changes at the formyl-NH terminus (in the membrane interior, away from the tryptophans), including fluorination of the formyl-NH-terminal valine, introduce voltage-dependent channel gating. Gramicidin channels are not just static conductors, but also dynamic entities whose structure and function can be manipulated by backbone and side chain modifications. PMID- 10472048 TI - Engineering charge selectivity in alamethicin channels. AB - The peptide alamethicin provides a system for engineering ion channel charge selectivity. To define alamethicin charge selectivity experimentally, we measured single-channel current-voltage relationships in KCl gradients using covalently linked peptide dimers. Two factors were found to contribute to the charge selectivity of these channels: (i) the ionic strength of the surrounding solutions; and (ii) the distribution of fixed charge on the peptide. Native alamethicin channels exhibited either cation selectivity or anion selectivity depending on which end of the channel was at the low salt side of the membrane. When the glutamine residue at position 18 in the sequence was replaced with a lysine residue, an anion-selective channel was obtained regardless of which end of the channel was at the low salt side of the membrane. PMID- 10472049 TI - Lorentzian noise in single gramicidin A channel formamidinium currents. AB - Seoh & Busath (1995) showed that in the presence of formamidinium, single gramicidin A channels were lengthened, had uniformly noisy currents at low voltages and had superlinear current-voltage relationships, all three properties being absent in gramicidin M- channels in which the interfacial tryptophan residues in gramicidin A are all replaced by phenylalanine. We measured the single channel noise power spectra (PSDs) in small monoolein (GMO) bilayers with formamidinium chloride solutions to help identify the mechanism of noise process. PSDs were Lorentzian with characteristic frequencies of 0.1-1.0 kHz in 0.1 and 0.3 M formamidinium chloride solutions, and from. 1-6 kHz in 1 M solution. Si(0), where measurable, ranged from approximately 50-200 fA2/Hz. The time course of the noise process could not be detected in these experiments. The low fc suggests slow motions or rare states of the blocking 'gates' which, judging from the result with gramicidin M-, must be equal to or related to the Trp residues. PMID- 10472050 TI - Can we use rate constants and state models to describe ion transport through gramicidin channels? AB - Can we use rate constants and state models to describe ion transport through gramicidin channels? Maybe, but only if rate constants are just proportionality constants between rates and probabilities of observing states of the channel. This approach is natural if the system of channel plus ions (plus water) is almost always in one or another of a small number of identifiable states. Many features of ion transport through gramicidin, including the conductance concentration relationship, concentration-dependent permeability ratios, anomalous mole fraction effect and to some extent flux ratio exponents, are consistent with a description in which there are four occupation 'states' of the pore: only water; an ion at one end; an ion at the other; and ions at both ends. Current-voltage relationships can (and must) also be fitted, but until there is a theory to predict the potential dependence of the rate constants this success will remain hollow. Other features have resisted interpretation. These include the failures to determine 'binding constants' consistent with all the data; the variation of flux ratio exponents with ion type; and, probably, the variation of the currents with asymmetrical ion concentrations. Nevertheless, state models still have one attractive feature, they allow consideration of the effects that one ion within the pore has on the movements of another. PMID- 10472051 TI - The binding site of sodium in the gramicidin A channel. AB - The available information concerning the structure and location of the main binding site for sodium in the gramicidin A channel is reviewed and discussed. Results from molecular dynamics simulations using an atomic model of the channel embedded in a lipid bilayer are compared with experimental observations. The combined information from experiment and simulation suggests that the main binding sites for sodium are near the channel's mouth, approximately 9.2 A from the centre of the dimer channel, although the motion along the axis could be as large as 1 to 2 A. In the binding site, the sodium ion is lying off axis, making contact with two carbonyl oxygens and two single-file water molecules. The main channel ligand is provided by the carbonyl group of the Leu10-Trp11 peptide linkage, which exhibits the largest deflection from the ion-free channel structure. PMID- 10472052 TI - The mechanism of channel formation by alamethicin as viewed by molecular dynamics simulations. AB - Alamethicin is a 20-residue channel-forming peptide that forms a stable amphipathic alpha-helix in membrane and membrane-mimetic environments. This helix contains a kink induced by a central Gly-X-X-Pro sequence motif. Alamethicin channels are activated by a cis positive transbilayer voltage. Channel activation is suggested to correspond to voltage-induced insertion of alamethicin helices in the bilayer. Alamethicin forms multi-conductance channels in lipid bilayers. These channels are formed by parallel bundles of transmembrane helices surrounding a central pore. A change in the number of helices per bundle switches the single channel conductance level. Molecular dynamics simulations of alamethicin in a number of different environments have been used to explore its channel-forming properties. These simulations include: (i) alamethicin in solution in water and in methanol; (ii) a single alamethicin helix at the surface of a phosphatidylcholine bilayer; (iii) single alamethicin helices spanning a phosphatidylcholine bilayer; and (iv) channels formed by bundles of 5, 6, 7 or 8 alamethicin helices spanning a phosphatidylcholine bilayer. The total simulation time is c. 30 ns. Thus, these simulations provide a set of dynamic snapshots of a possible mechanism of channel formation by this peptide. PMID- 10472053 TI - The Poisson-Nernst-Planck model. PMID- 10472054 TI - Ionic interactions in multiply occupied channels. AB - A significant number of physiologically important ion channels function via multi ion mechanisms where repulsion between ions at slightly separated locations is believed to be critical for permeation. We apply the semi-microscopic Monte Carlo approach and analyse how multiple occupancy affects permeation energetics and ion water-peptide correlations. We consider double occupancy in idealized models of two systems: gramicidin A and the KcsA K+ channel. We focus on the excess repulsion energy due to ion-water and ion-peptide correlations (repulsion energy adjusted for direct ion-ion interaction). Gramicidin, where multiple occupancy is marginally important functionally, is ideal for correlating structure and ion interactions. Pair occupancy is stabilized by interaction with bulk solvent, destabilized by interaction with both the channel water and, as binding sites are far apart, the peptide backbone. In the KcsA K+ channel, double occupancy is promoted by the uneven spacing and the large ion-water separations in the selectivity filter. The carbonyls forming the binding cavities are equally important for pair stabilization. Due to the binding pocket's design, net ionic repulsion is approximately 25-30% of what it would be in a gramicidin-like structure with the same interionic spacing. PMID- 10472055 TI - Peptide influences on lipids. AB - The extent to which the length of the membrane-spanning part of intrinsic membrane proteins matches the hydrophobic thickness of the lipid bilayer may be an important factor in determining membrane structure and function. To gain insight into the consequences of hydrophobic mismatch on a molecular level, we have carried out systematic studies on well-defined peptide-lipid complexes. As model peptides we have chosen gramicidin A and a series of artificial hydrophobic alpha-helical transmembrane peptides that resemble the gramicidin channel. These peptides consist of a hydrophobic stretch of alternating leucine and alanine residues with variable length, flanked by tryptophan residues. Using wide-line NMR techniques, we have investigated the interaction of these peptides with the bilayer-forming diacyl phosphatidylcholines and with phospholipids which by themselves have a tendency to form non-bilayer structures. We have shown that hydrophobic mismatch leads to systematic changes of the bilayer thickness and that it can even change the macroscopic organization of the lipids. The type of lipid organization induced by the peptides and the efficiency of the various processes depend on the properties of the lipids and on the precise extent of mismatch. PMID- 10472056 TI - Peptide-lipid interactions and mechanisms of antimicrobial peptides. AB - Hydrophobic matching, in which transmembrane proteins cause the surrounding lipid bilayer to adjust its hydrocarbon thickness to match the length of the hydrophobic surface of the protein, is a commonly accepted idea. To test this idea, gramicidin was embedded in dilauroyl phosphatidylcholine (DLPC) and dimyristoyl phosphatidylcholine (DMPC) bilayers at the molar ratio 1:10. The bilayer thickness (PtP) was measured by X-ray lamellar diffraction. In the fluid phase near full hydration, PtP is 30.8 A for pure DLPC, 32.1 A for DLPC/gramicidin mixture, 35.3 A for pure DMPC and 32.7 A for a DMPC/gramicidin mixture. Gramicidin apparently stretches DLPC bilayers and thins DMPC bilayers toward a common thickness as expected by hydrophobic matching. Gramicidin pair correlations were measured by X-ray in-plane scattering. In the fluid phase, the gramicidin-gramicidin nearest-neighbour separation is 26.8 A in DLPC bilayers but shortens to 23.3 A in DMPC bilayers, thus confirming the conjecture that when proteins are embedded in a membrane, hydrophobic matching creates a strain field in the lipid bilayer that in turn gives rise to a membrane-mediated attractive potential between proteins. These results were analysed with an elasticity theory of membrane deformation. The same principle explains the 'concentration-gating' mechanism of pore formation by antimicrobial peptides via the membrane-thinning effect. Concentration-gated pore formation and membrane thinning by alamethicin and magainin have been observed. PMID- 10472057 TI - Folding patterns of membrane proteins: diversity and the limitations of their prediction. AB - Significantly more high resolution structures of membrane proteins, obtained either by X-ray analysis, electron crystallographic methods or, in the future, by NMR spectroscopy, will be required for reliable structure predictions. Aberrations from the motifs of alpha-helical bundles and beta-barrels occur that are not easily identified by algorithms unless structural homologies exist in the data banks. The coexistence of secondary structure motifs, originally proposed for a neurotransmitter receptor, has now been confirmed for a bacterial iron siderophore translocating protein (FhuA) by X-ray analysis to 2.7 A resolution. This protein contains a plug domain that has both alpha- and beta-structures. PMID- 10472058 TI - Molecular basis of the charge selectivity of nicotinic acetylcholine receptor and related ligand-gated ion channels. AB - Nicotinic acetylcholine receptors are homo- or heteropentameric proteins belonging to the superfamily of receptor channels including the glycine and GABA A receptors. Affinity labelling and mutagenesis experiments indicated that the M2 transmembrane segment of each subunit lines the ion channel and is coiled into an alpha-helix. Comparison of the M2 sequence of the cation-selective alpha 7 nicotinic receptor to that of the anion-selective alpha 1 glycine receptor identified amino acids involved in charge selectivity. Mutations of the alpha 7 homo-oligomeric receptor within (or near) M2, namely E237A, V251T and a proline insertion P236' were shown to convert the ionic selectivity of alpha 7 from cationic to anionic. Systematic analysis of each of these three mutations supports the notion that the conversion of ionic selectivity results from a local structural reorganization of the 234-238 loop. The 234-238 coiled loop, previously shown to lie near the narrowest portion of the channel, is thus proposed to contribute directly to the charge selectivity filter. A possible functional analogy with the voltage-gated ion channels and related receptors is discussed. PMID- 10472059 TI - The gramicidin-based biosensor: a functioning nano-machine. AB - Biosensors combine a biological recognition mechanism with a physical transduction technique. In nature, the transduction mechanism for high sensitivity molecular detection is the modulation of the cell membrane ionic conductivity through specific ligand-receptor binding-induced switching of ion channels. This effects an inherent signal amplification of six to eight orders of magnitude, corresponding to the total ion flow arising from the single channel gating event. Here we describe the first reduction of this principle to a practical sensing device, which is a planar impedance element composed of a macroscopically supported synthetic bilayer membrane incorporating gramicidin ion channels. The membrane and an ionic reservoir are covalently attached to an evaporated gold surface. The channels have specific receptor groups attached (usually antibodies) that permit switching of gramicidin channels by analyte binding to the receptors. The device may then be made specific for the detection of a wide range of analytes, including proteins, drugs, hormones, antibodies, DNA, etc., currently in the 10(-7)-10(-13) M range. It also lends itself readily to microelectronic fabrication and signal transduction. By adjusting the surface density of the receptors/channel components during fabrication, the optimum sensitivity range of the device may be tuned over several orders of magnitude. PMID- 10472060 TI - Protons. PMID- 10472061 TI - Summary: what we have learned about gramicidin and other ion channels. PMID- 10472062 TI - Uterine emergencies. Atony, inversion, and rupture. AB - Uterine atony, inversion, and rupture are potentially fatal events that may occur in pregnancy. They are obstetric emergencies that require immediate attention. Although all women may experience these complications, identification or known risk factors allow the obstetric team to prepare for rapid diagnosis and intervention. This article includes management options to help prepare for these uncommon events. PMID- 10472063 TI - Amniotic fluid embolism. AB - Amniotic fluid embolism occurs rarely but is one of the leading causes of maternal mortality in the United States. The risk of death associated with this syndrome is 60% to 80% with half of survivors suffering long-term neurologic disability. The pathophysiology of amniotic fluid embolism is poorly understood. A review of the largest case series to date concluded that the physiologic and hematologic manifestations bear a greater resemblance to septic and anaphylactic shock than to any embolic phenomenon. Care of the patient who suffers amniotic fluid embolism is supportive. To date, no therapeutic interventions have been found to improve survival. PMID- 10472064 TI - Shoulder dystocia: an obstetric emergency. AB - Shoulder dystocia is one of the most dreaded complications of vaginal delivery encountered by the obstetrician. Although risk factors for shoulder dystocia exist, approximately 50% of cases do not demonstrate the classic predisposing signs. Obstetricians can help patients decrease their risk for fetal macrosomia by frequent attention to weight gain, nutrition, and exercise during pregnancy and by aggressive management of diabetes. All obstetricians must be familiar with the maneuvers used to effect delivery of impacted shoulders and must be prepared to institute these maneuvers immediately in a crisis situation. PMID- 10472065 TI - Trauma in the obstetric patient. AB - Trauma is the leading nonobstetric cause of maternal death. Penetrating trauma during pregnancy primarily involves gunshot and stab wounds. The incidence of visceral injury in pregnant women with penetrating abdominal trauma is 16% to 38% versus 80% to 90% in the general population. Management of pelvic trauma in pregnancy involves assessment of two individuals and decision making that is by necessity centered around preservation of the mother, avoiding ill-advised premature delivery of the fetus except in certain situations. PMID- 10472066 TI - Management of injuries to the urinary and gastrointestinal tract during cesarean section. AB - Proper management of surgical injuries occurring during cesarean section is important to minimize patient morbidity. Damage to the bladder is usually easy to identify and repair. The defect is closed with two or three layers of absorbable suture, and an indwelling catheter is left in place for 1 week. The management of ureteral injuries depends on their nature, extent, location, and time of discovery; consultation with a urologist is advisable. Small, well-perfused injuries to both the small and large bowel can be repaired primarily. More extensive bowel damage may require resection. Colostomy is no longer mandatory for patients with injuries to unprepared large bowel. PMID- 10472067 TI - Diabetic ketoacidosis in pregnancy. AB - Diabetic ketoacidosis (DKA) remains a medical emergency with high maternal and fetal mortality. Prompt recognition and resuscitative therapy markedly improves outcome. The pathophysiology and management of DKA in pregnancy is discussed in detail in this article. PMID- 10472068 TI - The prevention and management of eclampsia. AB - Hypertensive disease remains second only to embolic phenomena as a leading cause of maternal mortality. This article covers the major physiologic and pathologic findings to be considered when managing pregnant women with eclampsia. Attention to detail and an increased degree of suspicion will improve fetal and maternal outcomes. PMID- 10472069 TI - Atlas of obstetric and gynecologic emergencies. AB - Obstetric emergencies are rarely photographed. They often occur late at night when all personnel are involved in life-saving maneuvers, precluding the opportunity to record graphic representation of the pathology. Included in this atlas are several such cases. PMID- 10472070 TI - Cardiovascular risks related to increased diastolic, systolic and pulse pressure. An epidemiologist's point of view. AB - Since the introduction of the sphygmomanometer at the beginning to the 20th century, the significance of diastolic (DBP), Systolic (DBP) and pulse pressure (PP) as hypertensive cardiovascular risk factors has been controversial. These historical controversies are reviewed. Initially, DBP was thought to be the best measure of risk, but more recently both SBP and DBP, which ever is higher, are used in classifying hypertensive cardiovascular risk. There are problems with the present guidelines, in that SBP and DBP represent only two inflection points on the propagated pulse wave that is measured by cuff readings at the peripheral brachial artery. The heart is exposed to the central aortic pressure not to the brachial artery pressure. Moreover, both peripheral vascular resistance and large artery stiffness contribute to hypertensive cardiovascular risk. In middle-aged and elderly, elevated SBP is a better surrogate measurement of resistance than DBP, but SBP underestimates large artery stiffness. PP, the difference between peak SBP and end DBP, is the single best blood pressure surrogate for large artery stiffness. Epidemiological studies over the past decade point to SBP and DBP as the best cardiovascular risk markers for young subjects, whereas PP takes over as the more powerful risk marker for middle-aged and elderly subjects. These findings support the concept that cardiovascular events are more related to the pulsatile stress of large artery stiffness during systole than the steady-state stress of small vessel resistance during diastole. Therefore, at similar elevations of SBP, subjects with isolated systolic hypertension are at greater risk for cardiovascular events than those with combined systolic/diastolic hypertension. PMID- 10472071 TI - Epidemiology of the arterial stiffness. AB - Aortic stiffening is as much an important risk factor in cardiovascular morbidity and mortality, as it serves as reliable surrogate marker for clinical endpoints like myocardial and cerebrovascular incidents. Elevated aortic stiffness induces high systolic blood pressure, augmented pulse pressure with increased ventricular afterload, reduced subendocardial blood flow and augmented pulsatile stress in the peripheral arteries. Factors with relevant impact on the epidemiology of arterial stiffness are widely spread. 3 major groups of parameters influencing the stiffness of the aorta and the large arteries have been studied and described up to now: (i) physiological properties like age, gender, body height, pressure, hormonal state, genetic factors; (ii) environmental factors like nutrition (fish , salt-, garlic consumption), smoking, performance of sports and aerobic capacity; (iii) diseases like hypertension, hypercholesterolemia, diabetes, coronary heart disease, cerebrovascular disease, renal failure, Marfan-syndrome, growth hormone deficiency. Close association between several of these factors impedes analyzing them independently from each other. Age and blood pressure were found to be the most prominent predictors of arterial stiffness in normal as well as in disease populations. Physiological and environmental factors can modulate these effects of aging, diseases generally seem to amplify them. PMID- 10472072 TI - [Rigidity of large arteries and cardiovascular risk. epidemiological aspects and genetic determinants]. AB - Most of the morbid events due to hypertension and other risk factors are related to alterations of the large arteries of the brain, the heart or the kidney. Historically large arteries have been considered as passive conduits of blood, and physicians, surgeons and pathologists were mainly interested on their anatomical lesions such as rupture, stenosis, aneurysm, or thrombosis. However we know that large arteries are not passive conduit tubes but are characterized by elastic properties and are able to synthesize many vasoactive substances. These properties make the arterial wall a major modulator of the blood pressure and more generally of the cardiovascular regulation. Aging, environmental and genetic factors are responsible for structural and functional changes of the arterial wall media (hypertrophy, extracellular matrix accumulation, calcium deposits) and of the vascular endothelium (decrease in the release of vasodilators and increased synthesis of vasoconstrictors), all that leading to a diminution of elasticity and increased stiffness. The alteration of large arteries elasticity has deleterious effects on the heart upstream being responsible for an inadequate increase in systolic pressure and a relative decrease in aortic diastolic pressure at any given value of mean arterial pressure. The elevation in systolic pressure causes a disproportionate increase in end-systolic stress, which is the principal hemodynamic factor which promotes the development of cardiac hypertrophy, increased ventricular oxygen consumption, and left ventricular hypertrophy and can compromise capacity for coronary perfusion. Clinical and epidemiological studies have raised the possibility that subjects with stiffer arteries have wide pulse pressure, and that stiffening of large arteries is associated with excess morbidity and mortality independently of mean blood pressure. In addition to its etiologic role in cardiovascular disease, increased arterial stiffness may serve as an early marker for the diagnosis of asymptomatic atherosclerotic lesions, or for the evaluation of the severity of these lesions. In this review we report data from clinical, epidemiological and genetic studies, suggesting that arterial stiffness may be considered as a significant marker and/or an independent cardiovascular risk factor. This new concept should lead physicians to evaluate arterial stiffness for the prognosis and treatment of cardiovascular patients. PMID- 10472073 TI - Mechanical principles. Arterial stiffness and wave reflection. AB - Arterial (and predominantly aortic) stiffening with age is now acknowledged as the cause of isolated systolic hypertension, and the predominant cause of cardiac failure in the elderly. Aortic stiffening is gauged clinically from increase in brachial pulse pressure, but this underestimates change with age, since aortic pulse pressure increases far more than brachial (on account of substantial amplification of the peripheral arterial pressure pulse in young adults). Aortic stiffness can be measured as pulse wave velocity, but this too underestimates ill effects on the heart and central vessels, since the direct effect is amplified by early return of wave reflection. Ill effects of arterial stiffening can best be assessed through analysis of pressure wave contour from the carotid or radial site. Exploitation of relatively constant brachial transfer function enables the central aortic pressure wave to be synthesised from the radial pulse. This new clinical approach links traditional sphygmography (originally introduced in France) with conventional cuff sphygmomanometry, and is being evaluated in clinical and epidemiological studies. PMID- 10472074 TI - Mechanics of the large artery vascular wall. AB - The mechanics of large artery walls make a large contribution to the transmission toward regional vascular beds of energy generated by the left ventricle. It has been established that abnormal stiffness of the aorta and large conduit arteries is responsible in the short term for left ventricular overload followed by left ventricular hypertrophy. Radial and axial stresses imposed on vessel walls are defined in this review article. Mechanical parameters that can be measured in vivo and in vitro are described and compared; they include compliance, distensibility, elasticity modulus, impedance, and pulse wave velocity. PMID- 10472075 TI - Role of total arterial compliance and peripheral resistance in the determination of systolic and diastolic aortic pressure. AB - The goal of the study was to define the major arterial parameters that determine aortic systolic (Ps) and diastolic (Pd) pressure in the dog. Measured aortic flows were used as input to the two-element windkessel model of the arterial system, with peripheral resistance calculated as mean pressure over mean flow and total arterial compliance calculated from the decay time in diastole. The windkessel model yielded an aortic pressure wave from which we obtained the predicted systolic (Ps, wk) and diastolic (Pd, wk) pressure. These predicted pressures were compared with the measured systolic and diastolic pressures. The measurements and calculations were carried out in 7 dogs in control conditions, during aortic occlusion at four locations (the trifurcation, between trifurcation and diaphragm, the diaphragm and the proximal descending thoracic aorta) and during occlusion of both carotid arteries. Under all conditions studied the predicted systolic and diastolic pressure matched the experimental ones very well: Ps, wk = (1.000 +/- 0.0055) Ps with r = 0.958 and Pd, wk = (1.024 +/- 0.0035) Pd with r = 0.995. Linear regression for pulse pressure gave PPwk = (0.99 +/- 0.016) PP (r = 0.911). We found the accuracy of prediction equally good under control conditions and in presence of aortic or carotid artery occlusions. Multiple regression between pulse pressure and arterial resistance and total arterial compliance yielded a poor regression constant (r2 = 0.19) suggesting that the two arterial parameters alone cannot explain pulse pressure and that flow is an important determinant as well. We conclude that, for a given ejection pattern (aortic flow), two arterial parameters, total arterial resistance and total arterial compliance are sufficient to accurately describe systolic and diastolic aortic pressure. PMID- 10472076 TI - [Hypertrophy of the arterial wall and arterial rigidity]. AB - The purpose of this article is to review the relations between hypertension induced hypertrophy and stiffness of the arterial wall. Arterial remodeling was assessed based on the geometrical (lumen and arterial wall thickness) and functional (distensibility and Young's elastic modulus) properties of large and medium-sized arteries in hypertensive and normotensive subjects. Several clinical and animal studies have shown that arterial wall hypertrophy, which normalizes circumferential wall stress, does not increase the elastic modulus of arterial wall material during chronic essential hypertension. The structural changes associated with hypertension and those associated with aging have opposite effects on arterial distensibility at a given transmural pressure (isobaric distensibility): arterial distensibility increases with hypertension and decreases with aging. Thus, hypertension cannot be considered equivalent to aging. The structural and functional changes in arterial wall material associated with hypertension-induced hypertrophy may explain why medium-sized arteries maintain their distensibility characteristics despite a distending pressure increase; these changes may not be associated with an intrinsic (isobaric) decrease in the distensibility of large arteries. PMID- 10472077 TI - Terminology for the definition of arterial elastic properties. AB - A profusion of terminology for the description of arterial elastic properties can be found in the literature. This can cause confusion and make comparisons between studies and between different research groups problematical. The multitude of definitions--including "lay" terms--for indices like "compliance" and "distensibility" is highlighted, and it is proposed that a rapidly expanding field such as that studying arterial biophysical properties in vivo will not receive the respect it deserves if fundamental terms can have a multitude of different definitions, absolute values and units. It is also demonstrated that the beta stiffness index, quite widely quoted as a blood pressure corrected index, does nevertheless exhibit considerable blood pressure dependence- especially in patients with cardiovascular risk factors and/or disease. It is questioned whether all of the 24 indices identified for this review really are needed. Furthermore it is suggested that while a number of the parameters may provide complimentary information--depending on which aspects of the vasculature are to be described--there is a very real need for some form of consensus or agreement about terminology for the definition of arterial biophysical properties. PMID- 10472078 TI - [Diagnostic imaging in clinical tumor staging]. AB - In order to evaluate the anatomic extension of neoplastic disease according to the TNM system sufficiently, inclusion of imaging techniques is absolutely necessary. In addition, decisions on further clinical processing are based on precise identification of primary tumor extent (T), condition of regional nodes (N) and possible presence of distant metastases (M). Breast cancer, lung cancer, colorectal cancer and prostate cancer represent the most common tumor entities. Within this context the importance of imaging techniques for diagnosis, prognosis, therapy and follow-up of patients with these malignancies is presented in this report. PMID- 10472079 TI - [Malignant tumors of the mouth cavity and oropharynx. Stage classification]. AB - Staging of malignant tumours of the oral cavity and the oropharynx not only requires far more than a basic knowledge of anatomy and the usual pathways of spread, but also a broad understanding of the diagnostic benefits of current imaging modalities. As radiology should never try to replace histology, the main aim should be precise prediction of tumour margins and differentiation of tumour from edema and posttherapeutic changes. Only then will imaging studies have a significant clinical impact. PMID- 10472080 TI - [Staging of bronchial carcinoma]. AB - Staging of any tumor, i.e. determination of the extent of the disease, serves to select the patients who might profit from curative surgical intervention or to define those patients with inoperable carcinomas who should be referred for other therapies, such as chemotherapy or irradiation. Furthermore, accurate staging is necessary for assessment of prognosis, for radiation therapy planning, and for differentiation of those with small-cell lung cancer or for follow-up examinations of small-cell lung cancer patients after during and after chemotherapy. The primary radiological staging and diagnostic modalities for assessment of bronchial carcinomas are computed tomography (CT) of the thorax including liver and adrenal glands, abdominal sonography, and bone scintigraphy. Magnetic resonance imaging (MRI) should be reserved for specific indications, e.g. infiltration of the chest wall or staging of patients with intolerance/allergy to intravenous contrast medium. The clinical value of nuclear medicine techniques, such as [18F]2-fluoride-2-desoxy-D-glucose positron emission tomography (FDG-PET) for evaluation of lymph nodes and distant metastases, In-111 octreotide/somatostatin receptor scans for staging of small-cell lung cancer, and thallium-201 SPECT are currently being assessed in numerous studies, although these techniques are already in routine use. In future these or nuclear medicine techniques, as well as techniques using molecular-based contrast material, especially for MR imaging, currently in experimental status, may yield serious potential for staging purposes. PMID- 10472081 TI - [Radiologic staging of thoracic and abdominal tumors in childhood]. AB - Thoracoabdominal tumors in childhood can arise from all organs and affect children from the neonate to the adolescent. Better prognosis of childhood tumors, due to better biological understanding and improvement of chemotherapy, require sufficient radiological staging. Sufficiency in radiological staging needs modern cross-sectional techniques with high resolution, good availability and lower radiation dose. In general computed tomography (CT) is being used for radiological staging; increasingly MR imaging is being used because of multiplanar imaging techniques. Replacement of invasive techniques such as myelography and lymphography and modern cross-sectional techniques create painless staging conditions. Nevertheless, scintigraphy will always be a major examination technique, especially for evaluation of metastases and postoperative control examinations. The most common thoracoabdominal tumors in childhood and their staging are described. PMID- 10472082 TI - [Staging of breast carcinoma. Value of diagnostic imaging]. AB - In women with breast cancer, staging of the tumor plays an important role and often is considered to be the most prognostic and therapeutic factor. The most widely used staging system is based on the pathologic TNM classification. In recent years conservative therapies such as preoperative chemo- or radiotherapy are used more often with limited tissue evaluation resulting in insufficient or even inaccurate staging data. Tumor size as well as nodal involvement is decisive in selecting a treatment method and assessing the response. Thus, more accurate preoperative in vivo staging is necessary. This article therefore gives an overview of the different imaging modalities (mammography, ultrasonography, magnetic resonance imaging, and nuclear medicine) used for breast cancer staging. Furthermore, results of these methods are compared critically with the pathohistologic staging. PMID- 10472083 TI - [Staging of carcinomas of the upper gastrointestinal tract. The current status of diagnostic imaging]. AB - Esophageal carcinoma: CT and endosonography are complementary and the most important imaging modalities at present for staging. After endoscopic and histological diagnosis, CT of the thorax and the abdomen is used. With the proof of local infiltration of a neighbouring organ or in the presence of distant metastasis, palliative therapy can be started. If CT is not conclusive or no local infiltration or distant metastasis is proven, endosonography should be performed. Gastric carcinoma: At present endosonography shows the highest accuracy for diagnosis of the T stage. For differentiation between T3 and T4 tumors the accuracy of CT is not sufficient to predict resectability. The N stage can be determined at present by no modality with sufficient accuracy. Distant metastasis can be diagnosed by CT with high sensitivity and specificity. Small bowel carcinoma: These rare tumors are diagnosed with high accuracy by enteroclysis, whereby the diagnosis takes place at a late stage due to the nonspecific clinical findings. PMID- 10472084 TI - [Preoperative staging of colorectal carcinoma]. AB - Carcinoma of the colorectum is one of the most frequent neoplasias, with an incidence of 40 in 100,000. For the effective use of new, differentiated, less invasive treatment options, exact preoperative staging of the tumor is essential. The introduction of endosonography in rectal tumor staging allows for exact differentiation of the rectal wall layers and thus of tumor stages 1-3 with median accuracy of 89%. Magnetic resonance imaging of the rectum, especially in double-contrast technique, can also be employed in high and stenosing tumors and leads to an average accuracy of 85% for the stages 1-4. Computed tomography is the method of choice in screening for metastases. In lymph node staging, all modalities show only moderate accuracy around 75%. PMID- 10472085 TI - [Radiologic staging of liver and pancreatic malignancies]. AB - Modern therapeutic strategies can improve survival of patients with pancreatic or liver malignancies. A prerequisite is thorough radiologic assessment of these patients. A variety of radiologic techniques are available, such as ultrasound, contrast-enhanced CT, CTAP, contrast-enhanced MRI, and endoscopic ultrasound. In this review the advantages and weaknesses of the different techniques are presented in the staging of patients with HCC, cholangiocarcinoma, suspected liver metastases, and pancreatic carcinoma. In an era of economic restraints, funds are not flowing freely anymore for radiologic studies. We have to consider not doing so many different examinations for each patient. We have to decide in the future which diagnostic test we want to perform for detection and staging of a particular disease. For liver and pancreatic malignancies it is likely to be MRI that will take this role. PMID- 10472086 TI - [Characterization of adrenal gland space-occupying lesions. Can diagnostic imaging replace biopsy?]. AB - This paper describes the diagnostic value of new imaging techniques in characterization of adrenal masses and evaluates the role of adrenal biopsy. For differentiation of benign and malignant adrenal lesions, sensitivities and specificities of 85-100% can be reached by unenhanced computed tomography (CT) methods, chemical shift techniques in magnetic resonance imaging (MRI), delayed enhanced CT techniques, and by evaluation of wash-out curves in contrast-enhanced CT. The diagnostic value of all CT techniques depends on threshold values. The value of dynamic contrast-enhanced MRI is under discussion and should be reevaluated by using delayed enhanced series. Biochemical and scintigraphic methods (NP 59 iodine iodomethyl-norcholesterol and MIBG meta-iodobenzylguanidine scintigrams) are valuable for the diagnosis of functional adrenal masses; however, they do not allow differentiation of benign and malignant tumors. According to excellent results of new imaging techniques in characterization of adrenal masses, the indications for fine-needle aspiration biopsy have already regressed, as have complications associated with this invasive technique. PMID- 10472087 TI - [Radiologic staging of renal cell carcinoma]. AB - The routine staging work-up for renal cancer includes a contrast-enhanced multiphasic spiral CT and a chest radiograph. If there is doubt regarding the presence and extent of (supradiaphragmatic) IVC thrombus, MR imaging should be performed. Dynamic contrast-enhanced MR imaging should be used in place of CT in any patient with severe renal dysfunction, symptomatic polycystic kidney disease, or a history of allergy to iodinated contrast media. Cavography is no longer needed in the era of (adaptive array detector) spiral CT and MR venography. PMID- 10472088 TI - [Staging of musculoskeletal tumors in diagnostic imaging]. AB - The staging of primary malignant bone and soft tissue tumors requires adherence to a unified staging system. The staging system of the American Joint Commission for Cancer, the Enneking system, and the Hajdu system are described in respect of the radiologic staging criteria. Plain film radiography can estimate the biological aggressiveness of a bone tumor and in many cases narrow the differential diagnosis. Computed tomography is most sensitive in depicting cortical erosions and calcified periosteal reactions as well as in screening for distant metastasis. Angiography is mainly employed for vascular tumors and for the monitoring of interventional procedures such as tumor embolization. For both bone and soft tissue tumors, magnetic resonance tomography is the method of choice for the staging of intra- and extramedullary tumor extension, for the differentiation of neighboring compartments, and in the detection of invasion of the regional neurovascular bundle. After administration of contrast agents, the mode of enhancement gives further information on the nature of the tumor. With consistent application of the staging criteria in preoperative imaging, limb saving therapy can be employed more often and the prognosis of the disease can be improved. PMID- 10472089 TI - [FDG-PET in oncology. Methodological principles and clinical applications]. AB - Clinical PET is now primarily used in oncological indications. Fluorine-18 fluorodeoxyglucose (FDG) shows a high uptake in a variety of malignancies and accounts for approximately 95% of the oncological PET studies. The first part of this review deals with theoretical and methodological aspects of FDG-PET. Possible sources of errors for the method are discussed briefly. Oncological indications for FDG-PET are highlighted according to the results of consensus meetings of experts from nuclear medicine and oncology, organized by the German Society of Nuclear Medicine. FDG-PET is mainly used in staging and restaging, demonstrating of tumour recurrence and therapy control. Main benefits of the method are functional imaging, better spatial resolution than conventional scintigraphy and the possibility to perform whole-body imaging easily. A brief outlook on future prospects for the application of PET in oncology is given. PMID- 10472090 TI - [Diagnostic imaging in staging of gynecologic carcinomas]. AB - The prognosis in patients with gynecologic cancers depends not only on the stage but also on a wide spectrum of other findings. Cross-sectional imaging modalities, including sonography, CT and MRI, have increasingly been used for optimal treatment planning in gynecologic cancers. Their staging criteria are based on the well-established FIGO staging system. CT and MRI compete with sonography, which plays a pivotal role in the evaluation of the female pelvis. This paper reviews the role of sonography, CT and MRI in the staging of gynecologic malignancies. It puts the emphasis on MRI, which has been established as imaging modality of choice in the preoperative evaluation of cervical and endometrial cancer, and which seems slightly superior to CT in the staging of ovarian cancer. PMID- 10472091 TI - [A rare sequela of cardiopulmonary resuscitation. Left-sided diaphragmatic rupture]. PMID- 10472092 TI - [Physical and technical principles of B-image ultrasound]. PMID- 10472093 TI - [Rinke-Trauhand expert assessment proves: rate setting by the German health insurance are not distributed equitably. Radiologists of the North Rhine health insurance will pay about 5,000 DM to support mandatory health insurance services in 1999. The expert report exceeds all apprehensions]. PMID- 10472094 TI - [Effect of digital imaging, internet and teleradiology on radiologic structures]. PMID- 10472095 TI - The influence of spinal canal narrowing and timing of decompression on neurologic recovery after spinal cord contusion in a rat model. AB - STUDY DESIGN: The effect of spinal canal narrowing and the timing of decompression after a spinal cord injury were evaluated using a rat model. OBJECTIVE: To evaluate whether progressive spinal canal narrowing after a spinal cord injury results in a less favorable neurologic recovery. Additionally, to evaluate the effect of the timing of decompression after spinal cord injury on neurologic recovery. SUMMARY OF BACKGROUND DATA: Results in previous studies are contradictory about whether the amount of canal narrowing or the timing of decompression after a spinal cord injury affects the degree of neurologic recovery. METHODS: Forty adult male Sprague-Dawley rats were equally divided into a control group, in which spacers of 20%, 35%, and 50% were placed into the spinal canal after laminectomy, and an injury group in which the spacers were placed after a standardized incomplete spinal cord injury. After spacer removal, neurologic recovery in both was monitored by Basso, Beattie, Bresnahan (BBB) Locomotor Rating Scale (Ohio State University, Columbus, OH) motor scores and transcranial magnetic motor evoked potentials for 6 weeks followed by histologic examination of the spinal cords. Subsequently, 42 rats were divided into five groups in which, after spacer placement, the time until decompression was lengthened 0, 2, 6, 24, and 72 hours. Again, serial BBB motor scores and transcranial magnetic motor evoked potentials were used to assess neurologic recovery for 6 weeks until the animals were killed for histologic evaluation. RESULTS: Spacer placement alone in the control animals resulted in no neurologic injury until canal narrowing reached 50%. All of the control groups (spacer only) exhibited significantly better (P < 0.05) motor scores compared with the injury groups (injury followed by spacer insertion). Within the injury groups the motor scores were progressively lower as spacer sizes increased from the no-spacer group to the 35% group. The results in the 35% and 50% groups were not statistically different. The results of the time until decompression demonstrated that the motor scores were consistently better the shorter the duration of spacer placement (P < 0.05) for each of the time groups (0, 2, 6, 24, and 72 hours) over the 6-week recovery period. Histologic analysis showed more severe spinal cord damage as both spinal canal narrowing and the time until decompression increased. CONCLUSION: The results in this study present strong evidence that the prognosis for neurologic recovery is adversely affected by both a higher percentage of canal narrowing and a longer duration of canal narrowing after a spinal cord injury. The tolerance for spinal canal narrowing with a contused cord appears diminished, indicating that an injured spinal cord may benefit from early decompression. Additionally, it appears that the longer the spinal cord compression exists after an incomplete spinal cord injury, the worse the prognosis for neurologic recovery. PMID- 10472096 TI - The stability of reconstruction methods after thoracolumbar total spondylectomy. An in vitro investigation. AB - STUDY DESIGN: After total spondylectomy, five types of spinal reconstruction techniques were compared biomechanically. OBJECTIVES: To evaluate the stability provided by five reconstruction methods after total spondylectomy. SUMMARY OF BACKGROUND DATA: Total spondylectomy presents a worst-case scenario for spinal reconstruction. However, few investigators have biomechanically investigated spinal reconstruction stability after total spondylectomy. METHODS: Eight human cadaveric spines (T11-L5) were used. After intact analysis, a total spondylectomy was performed at L2 and reconstructed using Harms titanium mesh (Depuy-Motech, Warsaw, IN) as an anterior strut. Anterior, posterior, or circumferential instrumentation techniques were then performed using the Kaneda SR and ISOLA pedicle screw systems (AcroMed Corp., Cleveland, OH) as follows: 1) anterior instrumentation at L1-L3 with multisegmental posterior instrumentation at T12-L4 (AMP), 2) anterior instrumentation at L1-L3 with short posterior instrumentation at L1-L3 (ASP), 3) anterior instrumentation at L1-L3 (A), 4) multilevel posterior instrumentation at T12-L4 (MP), and 5) short posterior instrumentation at L1-L3 (SP). Nondestructive biomechanical testing was performed under axial compression, flexion-extension, and lateral bending loading modes. RESULTS: Only circumferential instrumentation techniques (AMP, ASP) exhibited higher stiffness than the intact spine in all loading modes (P < 0.05). Short circumferential fixation provided more stability than did multilevel posterior instrumentation (P < 0.05). Multilevel posterior fixation provided more stiffness than did short posterior and anterior instrumentation alone (P < 0.05). CONCLUSIONS: Only circumferential fixation techniques provide more stability than the intact spine in all testing modes. Short circumferential instrumentation provides more stability than multilevel posterior instrumentation alone and requires fewer levels of spinal fusion. PMID- 10472097 TI - Lumbosacral chordoma. Prognostic factors and treatment. AB - STUDY DESIGN: Retrospective analysis. OBJECTIVES: To analyze the prognostic factors in patients with chordomas, the success of various treatments, the diagnostic value of open versus needle biopsy, the neurologic impairment after sacral nerve resection, and the clinical presentation and site of origin. SUMMARY OF BACKGROUND DATA: Staging of chordomas has not been of much value, compared with other bone tumors, because for chordomas, grade is similar, metastasis is infrequent at presentation, and the prognostic significance of size is uncertain. METHODS: A review of patients with chordoma from 1965 through 1996 found 23 cases (mean age of patients, 55 years). The mean follow-up was 84 months. Mean tumor size was 81 mm (range, 35-135 mm), location was lumbar (n = 6), S1 (n = 4), S2 (n = 3), S3 (n = 7), S4 (n = 2), and S5 (n = 1). RESULTS: No tumors were found in the higher sacrum (S1-S2) alone, without involvement of the lower sacrum. Survival analysis at 5 years showed overall survival (OS) 86%, continuous disease free survival (CDFS) 58%, and local recurrence-free survival (LRFS) 60%. The location of tumor, defined by highest level of involvement (lumbar vs. sacrum) was of prognostic significance for OS (P = 0.01; log-rank test), CDFS (P = 0.036), but not for LRFS (P = 0.189). Results of multivariate regression showed that location was significant for OS (P = 0.007), CDFS (P = 0.008), and LRFS (P = 0.001). For patients with positive margins (n = 16), initial radiation correlated with longer CDFS (P = 0.002; Mantel-Cox) and LRFS (P = 0.005, Mantel-Cox), but was not significant for OS (P = 0.41). For patients who received no radiation, a positive margin correlated with a shorter CDFS (P = 0.04), a trend to shorter LRFS (P = 0.08), but no difference in OS. Therefore, both a tumor-free margin and initial radiation correlated with a longer survival. No patients had urinary or bowel dysfunction when both S3 nerves were preserved. If one S3 nerve was preserved, 1 of 3 patients had partial urinary incontinence and 2 of 3 patients required bowel medications. If both S3 nerves were resected, all patients required intermittent urinary catheterization and bowel medications. If both S2 nerves were resected, there was complete urinary and bowel incontinence. CONCLUSIONS: The highest level of tumor involvement was prognostically significant for OS, CDFS, and LRFS. Radiation was of value when complete excision was not achieved. Bilateral S3 nerve preservation is necessary to ensure retention of normal urinary and bowel function. PMID- 10472098 TI - Urgent surgical stabilization of spinal fractures in polytrauma patients. AB - STUDY DESIGN: A prospective, longitudinal study of multiply injured patients treated with segmental instrumentation for spinal fractures with a minimum 2-year follow-up. OBJECTIVES: To determine whether urgent stabilization of spinal fractures in severely injured patients increases the risk of surgery compared with early treatment and historical results. SUMMARY AND BACKGROUND DATA: Opinion in clinical studies is divided about whether operative treatment offers an advantage over nonoperative treatment in isolated spine fractures. Concomitant trauma is rarely discussed relative to decision making or surgical timing. Urgent stabilization of long-bone fractures improves survival and outcome in polytrauma patients. To date, urgent treatment of spine fractures in polytrauma patients has not been considered in the literature. METHODS: Seventy-five consecutive patients treated with segmental instrumentation for spinal trauma were observed prospectively to assess perioperative and longterm outcome. Twenty-seven patients with severe polytrauma (injury severity score, > 26) were separately analyzed. Perioperative and postoperative results were analyzed relative to timing of surgery, injury severity score, and surgical approach. Urgent treatment was defined as that provided within 24 hours of the spinal injury, and early treatment was defined as that provided between 24 and 72 hours after injury. RESULTS: Twenty-five patients (93%) sustained two or more major injuries in addition to the spine fracture, and 17 of 27 (63%) had neurologic injury. The mean injury severity score approached or exceeded the LD50 (50% expected mortality) in each group--36.0 for the early-treatment group and 42.0 for the urgent group--but only one patient in each group died. There were no deep venous thromboses, pulmonary emboli, neurologic injuries, decubiti, deep wound infections, or episodes of sepsis in either group. Blood loss for anterior procedures was significantly higher in the urgent group, but estimated blood loss for posterior procedures was similar for both groups. At 49 months' mean follow up, no revisions were necessitated by the urgent spinal treatment. CONCLUSIONS: Urgent spinal stabilization is safe and appropriate in polytrauma patients when progressive neurologic deficit, thoracoabdominal trauma, or fracture instability increase the risks of delayed treatment. PMID- 10472099 TI - Maintaining lumbar lordosis with anterior single solid-rod instrumentation in thoracolumbar and lumbar adolescent idiopathic scoliosis. AB - STUDY DESIGN: A prospective radiographic evaluation of 20 consecutive patients with primary lumbar or thoracolumbar adolescent idiopathic scoliosis who were treated with anterior convex compressive single solid-rod spinal instrumentation and structural titanium mesh (Harms) cages. OBJECTIVES: To evaluate a lordosis preserving anterior single solid-rod instrumented fusion technique for these specific adolescent idiopathic curves. SUMMARY OF BACKGROUND DATA: Maintaining instrumented segmental lumbar lordosis after anterior fusion and instrumentation for thoracolumbar and lumbar curves has been difficult. Twenty consecutive patients who underwent anterior single solid-rod fusion, aged 18 or younger with a primary thoracolumbar or lumbar curve, were observed for preservation of lordosis for a minimum of 2 years. METHODS: All patients underwent an identical anterior surgical technique, involving discectomies and anulectomies of all convex discs, structural titanium mesh (Harms) cages placed in the anterior half of all disc spaces below T12, morselized rib autograft packed in all disc spaces to be fused and inside the cages, and anterior single solid-rod (5.0-mm or 5.5-mm diameter) convex compressive spinal instrumentation with appropriate lordotic rod contour and rod rotation as necessary. The anterior rod was placed just posterior to the cages to optimize lordotic contouring of the spine during compression. None of the patients was braced after surgery. The lowest instrumented vertebrae (LIV) were L2 (n = 3), L3 (n = 15), and L4 (n = 2), typically the lower end vertebra of the Cobb measurement. RESULTS: Measurements for the primary coronal Cobb before surgery, 1 week after surgery, and 2 years after surgery were 48 degrees, 11 degrees, and 12 degrees; for C7 plumb line deviation from the midline: 3.6 cm, 1.9 cm, and 1.2 cm; for lowest instrumented vertebra translation: 31 mm, 15 mm, and 15 mm; and for LIV tilt: 29 degrees, 6 degrees and 6 degrees, respectively. Sagittal measurements before surgery, 1 week after surgery, and 2 years after surgery were: T12-L2: -1 degree, -6 degrees, and -6 degrees; T12-LIV: -8 degrees, -13 degrees, -9 degrees; T12-S1: -61 degrees, -56 degrees, -60 degrees; and entire instrumented levels: -6 degrees, -9 degrees, and -6 degrees, respectively. Coronal plane correction improved: 75% in the primary Cobb, 66% in the plumb line, 50% in LIV translation, and 80% in LIV tilt. Sagittal plane alignment improved in T12-L2 lordosis (P < 0.01) with preservation of physiologic lordosis in the instrumented levels, T12-LIV, and T12-sacrum. There were no instrumentation failures, pseudarthroses, or reoperations. CONCLUSIONS: Coronal plane correction with preservation of thoracolumbar and lumbar lordosis 2 years after anterior convex compressive spinal instrumentation was accomplished using a lordotically contoured single solid rod with structural cages placed anteriorly in the disc spaces of patients with primary thoracolumbar or lumbar adolescent idiopathic scoliosis. PMID- 10472100 TI - Spontaneous lumbar curve coronal correction after selective anterior or posterior thoracic fusion in adolescent idiopathic scoliosis. AB - STUDY DESIGN: Retrospective review of anterior and posterior fusions for treatment of adolescent idiopathic thoracic scoliosis. OBJECTIVES: To evaluate both the instrumented thoracic and the spontaneous lumbar curve corrections after treatment of the primary thoracic scoliosis by either anterior or posterior fusion. SUMMARY OF BACKGROUND DATA: Recent reports of thoracic scoliosis fusions have concentrated on the thoracic correction obtained by posterior segmental instrumentation systems. Coronal decompensation occurring because of curve progression with imbalance of the unfused lumbar spine has also been investigated. No report comparing spontaneous lumbar curve response after selective anterior versus posterior thoracic scoliosis fusions are available. METHODS: One hundred twenty-three cases of primary thoracic-compensatory lumbar adolescent idiopathic scoliosis were treated by selective thoracic instrumentation and fusion with either an anterior (n = 70) or posterior (n = 53) single approach. Thoracic and lumbar Cobb measurements and lumbar apical translation parameters were assessed before surgery, 1 week after surgery, and 2 years after surgery on upright coronal radiographs. All patients had a minimum 2 year follow-up. RESULTS: At 2-year follow-up, the percentage of thoracic curve correction was superior for the anterior (58%) versus the posterior (38%) group (P < 0.05), whereas the spontaneous lumbar curve correction was also superior for the anterior (56%) group versus the posterior (37%) group for all curve types investigated (P < 0.05). Both treatment groups consistently improved lumbar apical positioning after the thoracic fusion procedure. CONCLUSIONS: Spontaneous lumbar curve correction occurs consistently after both selective anterior and posterior thoracic fusion implying intrinsic ability of the lumbar spine to follow thoracic spine correction. In the current study, using multisegmented hook rod systems posteriorly with intentional limitation of posterior thoracic correction to avoid decompensation, instrumented thoracic and spontaneous lumbar curve correction was statistically better after anterior thoracic instrumentation and fusion, with the results most dramatic for lumbar curve Type C (true King II curves). PMID- 10472101 TI - Spinal deformity, pulmonary compromise, and quality of life in osteogenesis imperfecta. AB - STUDY DESIGN: A cross-sectional radiologic and clinical study of patients with osteogenesis imperfecta. OBJECTIVES: To determine whether pulmonary compromise is more closely correlated with scoliosis, kyphosis, or chest wall deformity in the population with osteogenesis imperfecta, and to assess the impact of spinal deformity, chest wall deformity, and pulmonary function on quality of life. SUMMARY OF BACKGROUND DATA: The incidence of scoliosis in osteogenesis imperfecta is between 39% and 80%. Up to 60% of patients with osteogenesis imperfecta have significant chest wall deformities. Pulmonary compromise is the leading cause of death in adults with osteogenesis imperfecta. METHODS: Fifteen patients with osteogenesis imperfecta between the ages of 20 and 45 were evaluated with sitting or standing anteroposterior and lateral radiographs of the entire spine, pulmonary function testing, and a validated health self-assessment questionnaire (Short Form-36). Radiographs were evaluated for thoracic scoliosis, thoracic kyphosis, and chest wall deformity. Correlation analysis was performed. RESULTS: Thoracic scoliosis was strongly correlated with decreased predicted vital capacity (r = -0.76). Significant diminution in vital capacity below 50% occurred at a curve magnitude of 60 degrees. Kyphosis and chest wall deformity were not predictive of decreased pulmonary function. Physical health (PCS) was closely correlated with predicted vital capacity (r = 0.65; P < 0.01) and with scoliosis (r = -0.52; P < 0.05). CONCLUSIONS: Thoracic scoliosis of more than 60 degrees has severe adverse effects on pulmonary function in those with osteogenesis imperfecta. This finding may partly explain the increased pulmonary morbidity noted in adult patients with osteogenesis imperfecta and scoliosis compared with that in the general population. PMID- 10472102 TI - Correlation between curve severity, somatosensory evoked potentials, and magnetic resonance imaging in adolescent idiopathic scoliosis. AB - STUDY DESIGN: A prospective study in patients with adolescent idiopathic scoliosis of different clinical severity using whole-spine magnetic resonance imaging and somatosensory evoked potentials. OBJECTIVES: To correlate the presence of magnetic resonance imaging structural abnormalities with somatosensory evoked potential-detected functional disorders in the hind brain and spinal cord and the Cobb's angle in patients with adolescent idiopathic scoliosis and to compare the result with those in healthy matched control subjects. SUMMARY OF BACKGROUND DATA: Many different types of neurologic dysfunction have been reported in scoliosis. With the advent of magnetic resonance imaging, there are increased reports on the association of idiopathic scoliosis and syringomyelia, Chiari I malformation, or tonsillar ectopia. The actual link between structural and functional disorder in idiopathic scoliosis is, however, unclear. METHODS: Posterior tibial nerve evoked potentials and whole spine magnetic resonance imaging were performed in 36 healthy control subjects, 135 patients with adolescent idiopathic scoliosis with Cobb's angle less than 45 degrees, and 29 patients with Cobb's angle more than 45 degrees. RESULTS: Tonsillar ectopia or syringomyelia, detected by magnetic resonance imaging, or functional disturbance in the somatosensory pathway, detected by somatosensory evoked potentials, was found to be significantly more frequent in the group of patients with severe scoliosis curvature, with an incidence of 31% and 27.6%, respectively. Incidence of tonsillar ectopia was 33.3% in patients with abnormal somatosensory evoked potentials in contrast to the much lower incidence of 2.9% in patients with normal somatosensory evoked potentials. There was a significant structural and functional link. The incidence of syringomyelia in patients with and without tonsillar ectopia was 33.3% and 0.7%, respectively. CONCLUSIONS: In patients with adolescent idiopathic scoliosis with severe curve, the significant association with tonsillar ectopia and abnormal somatosensory function points to a neural origin. Disorders in the somatosensory function may be one of the mechanisms linking tonsillar ectopia to scoliosis. Somatosensory evoked potentials and magnetic resonance imaging may have important diagnostic and predictive value and may help in the management of adolescent idiopathic scoliosis. PMID- 10472103 TI - Efficacy of intraoperative monitoring for pediatric patients with spinal cord pathology undergoing spinal deformity surgery. AB - STUDY DESIGN: A retrospective study of 38 pediatric patients with spinal cord pathology who underwent corrective spinal deformity surgery from January 1989 through June 1998. OBJECTIVES: To report reliability and specificity in obtaining intraoperative data in this population. These data were compared with monitoring results obtained in a group of pediatric patients with idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Reports in the literature suggest intraoperative monitoring for patients with spinal cord pathology may be of limited value. No optimal monitoring protocol has been suggested for this population. METHODS: The study group consisted of 38 pediatric patients with a diagnosis of spinal cord pathology who underwent corrective spinal deformity surgery from January 1989 through June 1998. All patients had lower extremity function. Somatosensory and neurogenic motor evoked potentials were used to monitor neurologic status during surgery. These data were compared with data obtained in 429 pediatric patients with idiopathic scoliosis. Study patients were divided into Group I, those who had had spinal cord surgery (n = 20), and Group II, those who had not (n = 18). RESULTS: Somatosensory evoked potentials were obtained in 93.2% and remained consistent with baselines in 87.2% of the study group patients. Neurogenic motor evoked potentials were obtained in 50.8% of the study subjects and remained consistent in 76.6% of those cases. The false-positive rate was 27.1% in the study group, compared with 1.4% in the group with idiopathic scoliosis. The study group had no true-positive or false-negative findings. Group I data differed from Group II data. CONCLUSIONS: Intraoperative monitoring should be used in patients with spinal cord pathology who undergo surgery for spinal deformity. Monitoring should not miss a neurologic deficit but demonstrates greater variability, resulting in more frequent use of an intraoperative wake-up test. PMID- 10472104 TI - Patients' perceptions of overall function, pain, and appearance after primary posterior instrumentation and fusion for idiopathic scoliosis. AB - STUDY DESIGN: This outcome study used patients' responses to the Scoliosis Research Society Outcomes Instrument to discriminate among patients who had undergone surgery for correction of juvenile or adolescent idiopathic scoliosis. OBJECTIVES: To evaluate a surgically treated population by using the SRS Outcomes Instrument. SUMMARY OF BACKGROUND DATA: The Scoliosis Research Society outcomes instrument was developed to help evaluate patient-perceived outcomes after treatment for idiopathic scoliosis. It includes 24 questions designed to investigate seven domains. METHODS: Eligible patients underwent posterior surgery for the first time before their 21st birthdays. One surgeon performed the surgery at one medical center. Of 168 eligible patients, 121 (72%) completed the Scoliosis Research Society outcomes questionnaire. RESULTS: Females reported better outcomes in the function after surgery (P = 0.005) and self-image after surgery (P = 0.01) domains. Preoperative curve pattern comparison demonstrated a significant difference in self-image after surgery among four groups classified according to curve pattern. The thoracolumbar and lumbar group recorded image scores of 5, the highest possible score, 85% of the time. The King-Moe (KM) V group scored 5, 75% of the time; the KM I and II group 48%; and the KM III and IV group 46% (P = 0.0015). After eliminating confounding variables, it was found that white patients reported experiencing less pain in follow-up than did black patients (P = 0.0098). Results were also suggestive that less pain was associated with increased number of fused vertebrae (P = 0.027). CONCLUSIONS: The strongest predictors of self-perceived favorable outcome among patients were female sex and white race. It is also suggested that longer fusions to L1 through L3 lead to less perceived pain than with shorter fusions. PMID- 10472105 TI - Complications in the surgical treatment of pediatric high-grade, isthmic dysplastic spondylolisthesis. A comparison of three surgical approaches. AB - STUDY DESIGN: An analysis of consecutive pediatric patients treated surgically for high-grade spondylolisthesis by one of three surgical procedures with emphasis on complications and functional outcomes. OBJECTIVE: Complications, radiographic results and patient-assessed function, pain, and satisfaction were assessed among three surgical procedures. SUMMARY OF BACKGROUND DATA: The existing literature is in disagreement about whether it is better to fuse without instrumented reduction or to use instrumentation and reduce high-grade dysplastic spondylolisthesis. METHODS: Thirty-two patients had 37 surgical procedures for Meyerding Grade 3 or 4 isthmic dysplastic spondylolisthesis. Eleven patients were treated with an in situ L4-sacrum posterior fusion without decompression (Group 1), 7 had posterior decompression with posterior instrumentation and posterior fusion (Group 2), and 19 patients had reduction and a circumferential fusion procedure (Group 3). All patients had new radiographs taken at time of follow-up (average, 3.1 years; range, 2 years-10 years, 1 month) and completed a functional outcome questionnaire. RESULTS: The incidence of pseudarthrosis was 45% (5 of 11) in Group 1, 29% (2 of 7) in Group 2, and 0% (0 of 19) in Group 3. All seven who had pseudarthrosis had small L5 transverse process surface area (< 2 cm2; P = 0.004). Only one patient had a neurologic deficit (unilateral extensor hallucis longus weakness) at time of follow-up. There were no significant differences among the groups in function, pain, and satisfaction in patients in whom solid fusion was obtained, but the scores were highest in Group 3. CONCLUSIONS: In situ fusion surgery in patients with high-grade spondylolisthesis with small L5 transverse processes (surface area, < 2 cm2) results in a high rate of pseudarthrosis. Circumferential procedures result in the highest rate of fusion and are effective in achieving fusion in those patients with established pseudarthrosis. The use of long (> 60 mm) iliac screws bilaterally (n = 21) in addition to bicortical sacral screws (four-point sacral-pelvis fixation) along with anterior column fusion reduces the risk of instrumentation failure in a decompression and reduction procedure. Outcomes of function, pain, and satisfaction are excellent in those in whom fusion is achieved. The risks in circumferential fusion-reduction procedures are acceptable. PMID- 10472106 TI - Complications and predictive factors for the successful treatment of flatback deformity (fixed sagittal imbalance). AB - STUDY DESIGN: This is an analysis of consecutive cases of flatback deformity (fixed sagittal imbalance), treated by one of two surgeons at a university hospital. OBJECTIVE: To define factors that contribute to results with treatment of flatback syndrome, classify types of sagittal deformities, and discuss complications. SUMMARY OF BACKGROUND DATA: There are few reports that detail the results and complications of current instrumentation and osteotomy techniques for correction of fixed sagittal deformities. METHODS: Twenty-eight patients treated with osteotomies for sagittal imbalance were eligible for 2-year minimum follow up (average, 3.6 years). Patients were classified (segmental imbalance, Type 1; or global imbalance, Type 2) and evaluated by upright radiographs, chart review, and a questionnaire. RESULTS: Twenty-eight (100%) patients returned the questionnaire, and 28 had current radiographs. Five treatment groups were evaluated based on osteotomy type (anterior, posterior [Smith-Petersen], both, or pedicle subtraction) and use of anterior structural grafting. All patients were treated with modern bilateral hook-rod-screw constructs. Mean correction at the osteotomy levels was 25 degrees for Type 1 deformities and 30 degrees for Type 2 (P < 0.05). Sagittal correction averaged 6.6 cm in Type 2 deformities (P < 0.05). Questionnaire analysis showed a significant and persistent reduction in subjective pain level. There were seven patients with 11 total complications and no neurologic deficits. Associations among patients who were not satisfied with their results (n = 4) included insufficient sagittal correction (P = 0.045), pseudarthrosis (P = 0.045), coronal imbalance, and four or more medical comorbidities (P = 0.03). CONCLUSIONS: Satisfaction with the results of treatment may be reduced in patients with four or more major co-existent medical problems, insufficient sagittal correction, and resultant pseudarthrosis. PMID- 10472107 TI - Minimum 5-year results of degenerative spondylolisthesis treated with decompression and instrumented posterior fusion. AB - STUDY DESIGN: An analysis of consecutive cases of degenerative spondylolisthesis treated by one of two surgeons at a university hospital. OBJECTIVES: To assess at a minimum 5-year follow-up the complication rate, reoperation rate, radiographic results, and patient satisfaction with surgical treatment of lumbar degenerative spondylolisthesis by means of segmental posterior instrumented fusion with decompression. SUMMARY OF BACKGROUND DATA: No reports of minimum 5-year follow-up for surgical treatment of degenerative spondylolisthesis were found in the literature. METHODS: The potential study population consisted of 49 consecutive patients who had undergone no prior surgery for degenerative spondylolisthesis (average age, 66.7 years; range, 52.2-78.7 years) with mean follow-up of 6.5 years (range, 5-10.75 years) who were treated with decompression, autogenous iliac crest bone grafting, intertransverse process fusion, and segmental (pedicle screw) instrumentation. Eight patients had died; the remaining 41 were included in the study sample. Thirty-six (88%) of the 41 patients returned an outcome questionnaire and had current radiographs. RESULTS: There was one case of instrument failure (one broken screw with late fusion), and one superficial infection. There were no neurologic deficits, no pseudarthroses, no recurrent stenosis at the fused segment, and no progression of deformity at the fused level. Five patients had symptomatic adjacent level transition syndromes. There were seven additional currently asymptomatic radiographic transition syndromes. Segmental sagittal Cobb angles were maintained at the fused level (17.7 +/- 8 18.8 +/- 7 degrees). Eighty-three percent reported satisfaction with the procedure, 86% thought their back and leg pain was still significantly better than before surgery, and 77% would have the procedure again if needed. Poor satisfaction (n = 4) was associated with more than four medical comorbidities (P < 0.03). A significant number (12 of 49, 24%) of patients had died or were ill more than 5 years after surgery. CONCLUSIONS: Radiographic transition syndromes were common. Major complications (2%), implant failures (2%), and symptomatic pseudarthroses (0%) were low. PMID- 10472108 TI - The selection of fusion levels using torsional correction techniques in the surgical treatment of idiopathic scoliosis. AB - STUDY DESIGN: This is a retrospective, consecutive case series, with the index patient included. OBJECTIVES: To evaluate the evolution and effectiveness of instrumentation techniques designed to untwist the scoliosis deformity. SUMMARY OF BACKGROUND DATA: Three-dimensional studies of the idiopathic scoliosis deformity are consistent with the theory that the deformity or deformities evolve as an imperfect torsion or torsions. METHODS: From 1989 through 1995, 102 consecutive patients (84 females, 18 males) underwent surgery with increasing emphasis on torsional correction. One hundred patients (98%), with an average age of 14.3 years (range, 10.5-20.8 years), were observed for an average of 40 months (range, 24-81 months). The upper instrumented vertebra evolved to be the centered vertebra. The lower instrumented vertebra was chosen based on its ability to become horizontal on contralateral bend radiographs and was termed the caudal foundation vertebra. Because these techniques evolved over the first 3 years of the study period, a split analysis was performed to evaluate improvements in correction and correction maintenance over the course of the study. RESULTS: The average Cobb angle was 59 degrees before surgery, 18 degrees after surgery (69% correction), and 22 degrees (63% correction) at latest follow-up. A comparison of the first half of the series with the second half showed no significant demographic differences. Curve correction was significantly improved for King-Moe IIB (thoracolumbar-lumbar curve only), King-Moe III, and King-Moe V curve types in the second half of the series. In the last 4 years, curve correction at latest follow-up for King-Moe IIB curves was 61% for the thoracic curve and 65% for the thoracolumbar-lumbar curve. King-Moe III curves had a 68% correction, and King Moe V curves had a 50% high thoracic and a 72% thoracic curve correction. Thoraco lumbar, lumbar, and King-Moe I curves averaged 81% correction of the thoracolumbar-lumbar curve. The angle of thoracic curve inclination improvement at 1 year was maintained at latest follow-up. CONCLUSIONS: This method of selecting instrumentation levels while using torsional correction techniques is safe and reliable. The results were improved with the evolution of these techniques and appear to provide improved correction and correction maintenance compared with that of historical controls. PMID- 10472109 TI - Langerhans' cell histiocytosis of the spine. Analysis of twenty-three cases. AB - STUDY DESIGN: Retrospective review of clinical and radiologic data in four major tertiary referral centers. OBJECTIVES: To report clinical and roentgenographic findings, to evaluate the results of various treatment methods, and to propose a protocol for management. SUMMARY OF BACKGROUND DATA: Langerhans' cell histiocytosis of the spine is a rare condition, and therefore, appropriate management is still controversial. METHODS: Clinical and roentgenographic findings of 38 vertebral lesions of 23 children, with average follow-up of 5.4 years, were investigated. This is the most extensive report apparent in the literature to date. The results of treatment were assessed clinically and radiologically. Anterior vertebral body height was measured sequentially to evaluate reconstitution of the vertebral body. RESULTS: The last follow-up examination demonstrated no clinical evidence of disease in all patients, regardless of treatment method. Neurologic deficits developed in four patients, but they completely disappeared. Satisfactory restoration of height was demonstrated in all except five vertebrae: one that had collapsed maximally when the patient was more than 15 years of age and four that had been fused anteriorly or posteriorly. Unsatisfactory results were also seen in a patient with progressive scoliosis and in one with an irregular endplate with disc space narrowing. Both of these complications developed after curettage. CONCLUSIONS: For treatment of single or dual spinal lesions, observation with or without bracing seems to be sufficient. In patients with multifocal lesions, chemotherapy produces good results. For treatment of neurologic deficit, low-dose radiotherapy is favored. Patients who underwent surgery--especially curettage and anterior fusion--had the worst outcome. PMID- 10472110 TI - Nonoperative treatment of neck and arm pain. PMID- 10472111 TI - Comparative study of TLSO. PMID- 10472112 TI - Laminitis in the horse. AB - This article serves as an introduction to this issue on laminitis. As such, it contains the general perspectives and terminology that will be used in all subsequent articles. This article separates the clinical problem of laminitis into developmental, acute, subacute, and chronic phases and defines the criteria, duration, clinical goals, and implications of these phases. The basis for the significance of laminitis to the horse industry and the horseman is reviewed. Lastly, the organization of this issue is described. PMID- 10472113 TI - Historical perspectives on laminitis. AB - This article attempts to provide a historical perspective regarding equine laminitis. It is designed to cover, as completely as possible, the historical record of, and the research advances made, in regards to acute and chronic laminitis. With respect to the historical record, the names given to this disease, the postulated etiologies, and the various treatment protocols are discussed. This article demonstrates the historical longevity of this disease and establishes a background for the current understanding of the disease's pathologic mechanisms and treatments. PMID- 10472114 TI - Clinical presentation, diagnosis, and prognosis of acute laminitis. AB - The purpose of this article is to describe the clinical features present in the patient affected with acute laminitis. The variations in the stance and characteristic lameness of the acute disease are described in relation to the disease severity and limb involvement. The elements of establishing a diagnosis including clinical history, physical and radiologic evaluation, and differential diagnosis are discussed. Difficulties and criteria used in establishing a prognosis of the acute phase patient are defined. PMID- 10472115 TI - The pathophysiology of developmental and acute laminitis. AB - This review implies that although we know more regarding the enigma of developmental and acute laminitis today than previously, there is still more to investigate. As these investigations are conducted and interpreted, new and more effective preventive and therapeutic regimens are likely to be developed, tested, and made available. As this occurs, the impact of laminitis should undoubtedly decrease. Unfortunately, due to the lack of clinical symptoms in the developmental phase and the shortness of the acute phase, it is also evident that the two sequelae of acute laminitis, subacute and chronic laminitis, are likely to continue to pose a major problem for some time. PMID- 10472116 TI - The pharmacologic basis for the treatment of developmental and acute laminitis. AB - Each horse with laminitis is presented to the veterinarian at a different stage in progression of the condition and with varying severity. The pathogenic timing is often unknown and is difficult to determine. Because timing and severity are related to both the lesion's severity and responsiveness to treatment, these factors are critical to treatment selection and success. It is erroneous to assume that each horse with laminitis should receive each treatment. It is therefore important to ascertain as logically and objectively as possible the pathophysiological stage of development of each horse when it is examined. Therapeutic failure may result from inappropriate interpretation of the clinical signs and pathophysiologic condition of the patient in order to coordinate a treatment regimen. PMID- 10472117 TI - Treatment of acute laminitis. Supportive therapy. AB - Over the last decade, both the farriery and veterinary professions have greatly increased their collective experience and understanding of the treatment of acute laminitis. Many horses that would have not been considered candidates for treatment 10 to 15 years ago are now saved, and some progress to successful careers as athletes. Unfortunately, the difficulties over prediction of the course of the disease persist, which continues to complicate treatment decisions. By its nature, supportive therapy is designed to make the horse more comfortable and limit further laminar injury by countering adverse biomechanical forces. Therefore, the success of treatment depends on the degree of instability between the distal phalanx and the hoof when treatment is started as well as the effectiveness of medical and supportive therapy. PMID- 10472118 TI - Clinical presentation, diagnosis, and prognosis of chronic laminitis. AB - This article focuses on the initial assessment of the horse affected with chronic laminitis. Variations in the clinical presentation and primary considerations in making a differential diagnosis are included. The elements of a clinical history essential to sound, therapeutic management, and prognosis are summarized. The physical and radiographic assessment of the digital lesions and diagnostic approaches to the common systemic aspects of the disease are presented and discussed. PMID- 10472119 TI - The pathophysiology of chronic laminitis. Pain and anatomic pathology. AB - The potential pathologic manifestations of chronic laminitis are just as varied, and possibly more so, than the list of possible inciting agents of the disease itself. The extent to which rehabilitation and return to normal function can be attained, cannot always be accurately determined by physical examination. It should be remembered that significant physiologic and pathologic alterations occur in chronic laminitis; thus, even if radiographically the patient returns to a normal appearance, residual morphologic and structural defects are likely to remain. PMID- 10472120 TI - The digital pathologies of chronic laminitis. AB - This review indicates that the patient-to-patient uniqueness commonly seen in chronic laminitis represents the variable presence of the digital pathologies. Although some degree of mechanical failure is always present, the secondary metabolic and growth dysplasias, vascular pathologies, and sepsis may or may not be evident. The presence and severity of these pathologies appear to have a more significant impact on the prognosis of individual cases than does the displacement of the distal phalanx. It should be reiterated that it is often the combined presence of these individual pathologies that gives rise to the patient that is totally refractory to treatment. In the absence of these pathologies, many horses with significant displacement of the distal phalanx are not in pain and are not in need of treatment. It thus follows that a key to the improved rehabilitation of difficult patients is focusing research on the physiopathology and diagnosis of these nonmechanical problems. PMID- 10472121 TI - The mechanisms and consequences of structural failure of the foot. AB - Many of the difficulties in managing the horse with chronic laminitis are associated with the foot's mechanical failure. The occurrence of digital collapse cannot be viewed in isolation as the primary therapeutic focus. The circulatory, metabolic, and growth pattern changes induced by the foot's mechanical collapse often serve as major limiting factors to successful rehabilitation. From the discussion above, it is obvious that a significant amount of research is needed to better understand the biomechanical pathologies of the failed foot. PMID- 10472122 TI - Trimming and shoeing the chronically affected horse. AB - Several of the technical approaches applied to the foot overlap with regard to intent. Frog or solar support, for example, may be provided either to stabilize the distal phalanx within the hoof capsule or in an effort to unload regional pain arising from the solar surface of the foot. It is likewise obvious that some techniques such as lowering the heels to achieve phalangeal realignment and raising the heels to relieve deep digital flexor tendon tension are contradictory. In these instances, it is not that one technique is always correct but that differences exist among horses. Currently, it is something of an art to define what specific technique is needed or, alternatively, how to best apply a specific technique. As more facts regarding how the normal and foundered foot function, the farrier's role in the rehabilitation of affected horses is likely to increase. PMID- 10472123 TI - Laminitis as a systemic disease. AB - This article presents the clinical pathology and the involvement of the cardiovascular, renal, endocrine, and immunologic systems in laminitis. The data available on these systems are presented with respect to the disease phase and severity. The nutritional and metabolic alterations realized in the chronically affected horse are also presented. In this discussion, the origins and clinical implications of these systemic findings are discussed. PMID- 10472124 TI - Pharmacologic and alternative therapies for the horse with chronic laminitis. AB - This article deals with treatment of the chronically foundered horse. The first section of this article is focused on aspects of the traditional pharmacologic approaches to management of digital pain and sepsis, dietary management, and thyroid supplementation. A second section introduces the concepts, principles, and agents that are used in homeopathic treatments for laminitis. Lastly, a third section of this article reviews the use of acupuncture and traditional Chinese medicine approaches to treatment of chronic laminitis. PMID- 10472125 TI - [GP IIb/III antagonists for the optimization of fibrinolysis. Simple management through rapid start of action and easy manageability]. PMID- 10472126 TI - A World Wide Web-service to aid the development of AMBER parameters using analogy to standard parameters. AB - A World-Wide Web service has been constructed to assist the development of force field parameters extending the AMBER force fields. This service extracts parameters from the standard AMBER force field parameter databases. From a Web based interface the user can choose between bond, angle and torsional parameters with certain constraints on element and/or atom hybridization. The software constructed for the purpose of finding appropriate parameters will locate standard AMBER force field parameters matching the user specification. For bond and angle parameters a scatter plot of the reference values against force constants is provided. This service has been produced to assist in extraction and evaluation of parameters that may be useful for molecules other than proteins and nucleic acids. PMID- 10472127 TI - Influence of noradrenaline on the respiratory status of Rana balcanica red cell suspension under normoxia, hypoxia and hypercapnia: alpha 1-receptor involvement. AB - The effect of normoxia, hypoxia and hypercapnia on the extracellular pH, partial pressure carbon dioxide (pCO2), partial pressure oxygen (pO2) and HCO3- levels after noradrenaline treatment of Rana balcanica erythrocytes, was investigated. Noradrenaline caused a significant reduction of the extracellular pH which may have been due to the activation of red blood cell Na+/H+ exchange. Significant falls in the partial extracellular pressure of CO2 and O2 were evident. The initial reduction in extracellular pCO2 and pO2 was followed by a rise reflecting the desensitization of the Na+/H+ exchange after 15 min of hormone stimulation. Both hypercapnia and hypoxia increased the magnitude of these changes in relation to normoxia, although the greatest changes were observed under hypercapnic conditions. The involvement of alpha 1 receptors in regulating the concentration of respiratory gases after catecholamine stimulation was demonstrated. It is suggested that these responses increased the effectiveness of gas transfer over the respiratory surfaces. PMID- 10472128 TI - Resistance to antibiotics, metals, hydrophobicity and klebocinogeny of Klebsiella pneumoniae isolated from foods. AB - Milk samples and milk products (69 in toto) were screened for the presence of Klebsiella pneumoniae (52%), and maximum isolations (77%) were from ice cream samples (13). The isolates were hydrophobic, non-haemolytic and possessed both mannose resistant (MR) and mannose sensitive (MS) pili or only MR pili when tested with human or sheep blood, respectively. All isolates were resistant to one metal at least whereas about 98% exhibited resistance to two or more metal ions. The resistance frequency of 93%, 90% and 66.7% was observed against silver (20 micrograms/ml), cadmium (20 micrograms/ml) and mercuric ions (20 micrograms/ml), respectively. Multiple drug resistance (MDR) was observed in 10% of the isolates only. A direct correlation between the metal ion and antibiotic resistance was found in MDR strains. The klebocin typeability of 53% and 61% was observed with 153-158 and 153-156, U-5 and U-6 groups, respectively. The most common typing patterns involved strains 424 (21%) and 442 (31.8%). Only 61% of the isolates showed enterotoxigenicity by the coagglutination test. PMID- 10472129 TI - The potential for long-range transport of air-pollutants into Greece: a climatological analysis. AB - A flow climatology using 5 years (1983-1987) of back trajectories has been created for the Aliartos meteorological-chemical station (Greece's EMEP-station phi = 38 degrees 13'N, lambda = 23 degrees 00'E) using the Air Resources Laboratory (ARL) trajectory model. Air mass back trajectories were analysed to quantify the contributions of major source areas to the air pollution concentration in the area of Greece. The trajectories were produced two times a day at the 850-hPa and 700-hPa pressure surface. The trajectories were classified and summarised in order to produce a long range trajectory climatology. The purpose of this paper is to use the above flow climatology to improve understanding to atmospheric transport pathways to Greece (and to the greater south-eastern area of Europe) and to evaluate the polluting potential of distant source regions outside the country. The limited chemical concentration data available from Aliartos are used to evaluate the conclusions reached from the flow climatology and emissions analysis. The results show that trajectories reach Greece from five major source areas: (i) Northwest Europe, (ii) west Europe, (iii) north Europe, (iv) Northwest Asia, and (v) Northwest Africa. Sector analysis of air pollution concentrations and emissions also showed that: (i) Most air pollutants transport from distances beyond 1800 km takes place, and the most probable source areas are the industrialised countries of Europe. (ii) Countries of west and north-central Europe emit the greater amounts of air pollutants (S and N). (iii) Sulphate and nitrate concentrations are similar among transport sectors. PMID- 10472130 TI - Bioassay-directed fractionation of PAH of molecular mass 302 in coal tar contaminated sediment. AB - Bioassay-directed fractionation was used to characterise genotoxic polycyclic aromatic hydrocarbons (PAH) of molecular mass 302 amu in organic solvent extracts of coal tar-contaminated sediment from Sydney Harbour, Nova Scotia. A normal phase HPLC technique was employed to separate PAH-rich solvent extracts into fractions containing PAH of single molecular mass classes. The 302 amu molecular mass fraction was isolated and further separated using reversed phase HPLC; subfractions were collected every 30 s and subjected to bioassay analyses with Salmonella typhimurium strain YG1025 with the addition of oxidative metabolism (4% S9). Compounds eluting in the most active subfractions included naphtho[2,1 alpha]pyrene and naphtho[2,3-alpha]pyrene. The results of this study underscore the significant contribution that molecular mass 302 PAH make to the biological activity of complex environmental mixtures. PMID- 10472131 TI - Evaluation of cycling patterns of dissolved oxygen in a tropical lake as an indicator of biodegradable organic pollution. AB - Dissolved oxygen cycling patterns in a tropical lake (Weija Lake) were shown to be useful as a potential indicator of biodegradable organic pollution, by dosing with liquid malt to give an additional organic burden of 2.5 and 5.0 mg l-1 TOC, and monitoring the DO values continuously for 140 h. These loadings were added to water columns (in tubes) suspended from a raft in a lake in south-east Ghana. The addition of organic pollution burden to the lake water produced two main effects: the mean DO value was lowered, and the amplitude of the DO cycle decreased as organic loading increased from 2.5 to 5.0 mg l-1 TOC. There was also an indication of heterotrophic respiration associated with organic inputs for the 5.0 mg l-1 added TOC suggesting a P/R ratio of well below 1.0. Taking the results of a DO cycling computer model together with those from the lake raft experiments, it can be concluded that dissolved oxygen cycles can be a good indicator of biodegradable organic pollution load. PMID- 10472132 TI - Fluxes of radiocaesium in selected rural study sites in Russia and Ukraine. AB - Food production and food harvesting systems common in the areas contaminated by the Chernobyl accident in Russia and Ukraine can be grouped into three major categories: collective farm produce, private farming produce and foods collected from natural ecosystems. The contribution of each of these sources to radiocaesium intake by people living in rural settlements in the mid 1990s has been estimated at two major study sites, one in each country. The collective farm system provided the smallest contribution (7-14%) to the intake of radiocaesium at both sites. Natural food was the major contributor to intake at the Russian site (83%). Whereas private farm produce was the major contributor (68%) at the Ukrainian study site. The difference between the two sites was mainly because private milk production was stopped at the Russian site due to the contamination in 1986. A retrospective assessment of the situation 1 year after the accident shows that collective farming could have been a minor contributor to radiocaesium intake (8%), whilst private farming would have been the major contributor wherever private milk production and consumption continued. The extent to which inhabitants consume natural foods from forests has a considerable effect on their radiocaesium intake. The comparative importance of food products from natural ecosystems increases with time due to the long effective ecological half-lives of radiocaesium in unimproved pastures and forests. Estimation of the fluxes of radiocaesium from the different production and harvesting systems showed that the contribution from private farming and food harvesting from natural ecosystems may be significant, contributing 14-30% to the total fluxes of radiocaesium from an area even if the quantity of food produced in these systems is small. However, the major contributor to the flux exported from an area was the collective farming system, accounting for about 70-86% of the total. PMID- 10472133 TI - Agricultural pesticides threaten the ecological integrity of northern prairie wetlands. AB - The northern Great Plains of North America has millions of small wetlands, and these are often dispersed through cultivated fields. We investigated relationship between pesticide occurrence and precipitation in selected wetlands in a 30.4 x 10(4) km2 area of the Great Plains with relatively uniform farming practices and 1,777,600 wetlands (southern Saskatchewan, Canada). By early July after pesticides have been applied to crops, the mean number of pesticides detected in wetlands ranged from 1.8 in regions with little precipitation (< 21 mm rain during the previous 15 days) to 3.2 in regions under higher rainfall (> 90 mm). The proportion of wetlands in which at least one pesticide exceeded Canadian guidelines for the protection of aquatic life increased from 0% to 60% over this same precipitation range. The maximum number of pesticides detected in a single wetland was six. Concentration of lindane in wetlands increased with increasing precipitation. Using geographic information on rainfall, wetland densities, area seeded to crops, and region specific relationships between pesticides and precipitation, we estimated the number of wetlands in Saskatchewan with elevated levels of pesticides. In early July, during 3 of the 6 years, the number of wetlands subjected to pesticide levels that exceeded guidelines for the protection of aquatic life was significant, ranging from 152,000 to 424,000 wetlands or 9-24%, respectively, of the total. Lindane and triallate exceeded the guidelines most frequently. PMID- 10472134 TI - Factors affecting organochlorine contaminant concentrations in milk and blood of northern fur seal (Callorhinus ursinus) dams and pups from St. George Island, Alaska. AB - Levels of organochlorine contaminants in blood of northern fur seal (Callorhinus ursinus) pups and the blood and milk of their dams early in the lactation period are reported here. The contaminants included 15 selected individual polychlorinated biphenyl (PCB) congeners and DDT metabolites identified through high-performance liquid chromatography. Congeners CB-77, -81, -126, -169 and -189 were below the limits of detection in milk and blood samples analyzed. Congener specific concentrations of PCBs in the blood of pups were compared based on the age of their dam (< or = 5 years or > 7 years). Pups of young (presumably primiparous) dams had significantly elevated levels of CBs-101, -118, -128, -138, -153/87, -170/194, and -180 than pups of older (multiparous) dams. Congeners CB 128 and -170/194 were detected in the blood of pups of young dams but not in the blood of pups of older dams nor in any of the dams blood. Additionally, pups had higher blood levels in seven of 10 detected PCB congeners as compared to the levels measured in milk when adjusted for lipid content. Levels of DDT metabolites and toxic equivalency quotients of dioxin-like congeners followed similar trends. Lipid-normalized concentrations of CB-101 and total PCBs were significantly higher in the blood of dams than in their milk. CB-128, -156, -157, -170/194, p,p'-DDT, o,p'-DDT, p,p'-DDD and o,p'-DDD were not detected in dam blood samples, but were detected in milk samples. Calculation of 'biomagnification factors' from milk to pup blood indicated a biomagnification of CB-101, -105, -118, -138, -153/87, and -180. Significant mean accumulation factors ranged from 1.5 to 7.5. Inter-annual differences in exposure levels and specific congener concentrations in both milk and blood were apparent. Northern fur seal pups, especially first-born, have a substantial exposure to organochlorine contaminants at a critical developmental stage. PMID- 10472135 TI - Residues of organochlorine pesticides in fruits, vegetables and tubers from Nigerian markets. AB - Residue levels of organochlorine pesticides have been determined in raw fruits, vegetables and tubers from markets in Nigeria. In the fruits, total HCH, aldrin and total DDT were detected in 77, 38 and 30% of all samples, respectively. In the vegetables, total HCH, HCB, total DDT and aldrin were detected from 95, 53, 50 and 30%, respectively, of all samples. Aldrin + dieldrin, total HCH, and total DDT were detected from 98, 79 and 49%, respectively, of all tuber samples. Other pesticides were below their detection limits. The average levels were generally low and none were above the FAOs maximum residue limits. PMID- 10472136 TI - Metal and sediment ingestion by dabbling ducks. AB - The chemical analysis of intestinal digesta from hunter-killed carcasses or of wildlife scat is a promising means of estimating the exposure of wildlife to those environmental contaminants that, like lead, are poorly absorbed in the digestive tract. When evaluating contaminants at a site, biologists may find the results of this non-destructive approach more straightforward to interpret in terms of exposure to wildlife than would be analyses of soils, sediments, water, or wildlife tissues. To illustrate the approach, we collected digesta from 47 waterfowl shot by hunters at Prime Hook National Wildlife Refuge, in Delaware, USA. The waterfowl digesta contained an average of approximately 2.4% sediment, estimated from the Al concentrations in the digesta, a marker for sediment. Al concentrations were significantly correlated with concentrations of Cr (Spearman's rank correlation coefficient, r = 0.57), V (r = 0.70), Ni (r = 0.31), and Pb (r = 0.55), and we concluded that these metals were ingested mainly with sediment. American widgeon (Anas americana) ingested sediment at a rate of about four times that of three other species of dabbling ducks (Anas crecca, A. acuta, A. rubripes) and had several times the exposure to the sediment-associated metals. The digesta of one American black duck contained a high concentration of lead (70 mg/kg, dry wt.), presumably from lead shot, but none of the other samples had notably elevated metal concentrations. We suggest that scat and digesta be analyzed more widely by biologists and resource managers seeking a simple, inexpensive assessment of contaminants in local wildlife habitat. PMID- 10472138 TI - Medical etiquette: pro and con. PMID- 10472137 TI - Nuclear materials production in the former USSR. IAEA Bull. 40, p 11 (1998) PMID- 10472139 TI - The Dutchman who discovered a vast New World. PMID- 10472141 TI - Is it time to close the book on closed-book examinations? PMID- 10472140 TI - Brown University School of Medicine: Class of 1999. PMID- 10472142 TI - Charles Whitney Page, MD, and the abolition of restraints in American psychiatry. PMID- 10472143 TI - Bungee jumper's eye. PMID- 10472144 TI - Clinical performance measurement. PMID- 10472145 TI - Antiplatelet agents and the role of clopidogrel in the prevention of secondary atherosclerotic events. PMID- 10472146 TI - Use of Massachusetts hospitals by Rhode Island residents. PMID- 10472147 TI - Bicycle helmets: it's the law. PMID- 10472148 TI - The Good Samaritan physician. PMID- 10472149 TI - A multi-attribute model of prostate cancer patient's preferences for health states. AB - Multi-attribute utility theory (MAUT) provides a way to model decisions involving trade-offs among different aspects or goals of a problem. We used MAUT to model prostate cancer patients' preferences for their own health state and we compared this model to patients' global judgments of health state utility. 57 patients with prostate cancer (mean age = 70) at two Chicago Veterans Administration health clinics were asked to evaluate health states described in terms of five health attributes affected by prostate cancer: pain, mood, sexual function, bladder and bowel function, and fatigue and energy. Each attribute had three levels that were used to form three clinically realistic health state descriptions (A = high, B = moderate, C = low). A fourth personalized health description (P) matched the patient's current health. We first measured patients' preferences using time trade-off (TTO) judgments for the three health states (A, B, and C) and for their own current health state (P). The TTO for the patient's own health state (P) was standardized by comparing it to TTO judgments for states A and C. We next constructed a multi-attribute model using the relative importance of the five attributes. The MAU scores were moderately correlated with the TTO preference judgments for the personalized state (Pearson r = 0.38, N = 57, p < 0.01). Thus, patients' preference judgments are moderately consistent and systematic. MAUT appears to be a potentially feasible method for evaluating preferences of prostate cancer patients and may prove helpful in assisting with patient decision making. PMID- 10472150 TI - Reliability and validity of the Functional Assessment of Cancer Therapy Colorectal (FACT-C) quality of life instrument. AB - BACKGROUND: This paper describes the development of the Functional Assessment of Cancer Therapy-Colorectal (FACT-C)--a questionnaire assessing quality of life concerns pertinent to colorectal cancer patients. This self-report instrument combines specific concerns related to colorectal cancer with concerns that are common to all cancer patients as assessed with the FACT-General (FACT-G). METHOD: Three separate and distinct validation samples were used with different ethnic and stage of disease compositions. RESULTS: Results suggest that the FACT-C is a reliable and valid measure in both its English and Spanish language versions. Specifically, the FACT-C evidenced good internal consistency reliability and concurrent validity, as well as an ability to distinguish between groups based on functional status and extent of disease. The FACT-C was also found to be sensitive to changes in functional status. Colorectal cancer-specific items form the Colorectal Cancer Subscale (CCS) which was found to have adequate convergent and divergent validity. Internal consistency was adequate in most samples. In addition, the CCS was able to distinguish among groups that differed in functional status and was sensitive to change in functional status. CONCLUSION: It is recommended that the entire FACT-C (rather than simply the CCS) be used when studying patients with colorectal cancer in order to provide a comprehensive assessment of quality of life. Recommendations are offered concerning the appropriate use of these measures in clinical research and directions for future research with colorectal cancer patients. PMID- 10472151 TI - A comparison of two approaches for assessing patient importance weights to conduct an Extended Q-TWiST analysis. AB - OBJECTIVE: Patient-centered methods for evaluating treatments require validated preference-elicitation techniques. We describe the validation of two preference elicitation approaches for use in an Extended Q-TWiST treatment evaluation. The first method was an "idiographic" approach, which attempts to capture intra individual differences in the degree to which each domain distracted from and interfered with life activities. The second method, a Likert-scaled approach, asks patients to evaluate the importance of each quality-of-life (QOL) domain. METHODS: Patient-reported QOL and preferences were assessed in participants with gastroesophageal reflux disease at baseline (n = 172), one week (n = 25), and 4 weeks after baseline (n = 100). RESULTS: Both approaches demonstrated high internal consistency and the ability to discriminate known groups based on reported pain and number of days with symptoms. The idiographic approach exhibited responsiveness, although it was more highly correlated with QOL than the Likert-scaled approach. The Likert-scaled approach had good face validity but demonstrated low reliability compared to the idiographic approach. CONCLUSIONS: Both preference-elicitation methods exhibited promise as well as limitations. Future research should focus on increasing the reliability of the Likert-scaled approach, reducing the overlap between the idiographic approach and QOL, and examining the relationship between reliability and responsiveness for a range of illness trajectories. PMID- 10472152 TI - The Assessment of Quality of Life (AQoL) instrument: a psychometric measure of health-related quality of life. AB - This paper describes constructing the Assessment of Quality of Life (AQoL) instrument; designed to measure health-related quality of life (HRQoL), and to be the descriptive system for a multi-attribute utility instrument. Unlike previous utility instruments' descriptive systems, the AQoL's has been developed using state-of-the-art psychometric procedures. The result is a descriptive system which emphasizes five different facets of HRQoL and which can claim to have construct validity. Based on the WHO's definition of health a model of HRQoL was developed. Items were written by focus groups of doctors and the researchers. These were administered to a construction sample, comprising hospital patients, and community members chosen at random. Final construction was through an iterative process of factor and reliability analyses. The AQoL measures 5 dimensions: illness, independent living, social relationships, physical senses and psychological wellbeing. Each has three items. Exploratory factor analysis showed the dimensions were orthogonal, and each was unidimensional. Internal consistency was alpha = 0.81. Structural equation modeling explored its internal structure; the comparative fit index was 0.90. These preliminary results indicate the AQoL has the prerequisite qualities for a psychometric HRQoL instrument for evaluation; replication with a larger sample is needed to verify these findings. Scaling it for economic evaluation using utilities is being undertaken. Respondents have indicated the AQoL is easy to understand and is quickly completed. Its initial properties suggest it may be widely applicable. PMID- 10472153 TI - The development and validation of an instrument to assess acute sinus disease in children. AB - Our goal was to produce a reliable, responsive instrument to quantify disease burden in children with acute sinusitis for use in clinical trials. In a cross sectional survey of 1611 community pediatric patients, parents rated the burden attributable to 13 sinus symptoms. Using logistic regression, we identified five symptoms that predicted the clinical diagnosis of sinusitis. The S5 is the average symptom score for nasal obstruction, daytime and nighttime coughing, headache and colored nasal discharge (range 0-3). The S5 was high in children with acute sinusitis (mean = 1.54, SD = 0.77, N = 93), and low in well children (mean = 0.42, SD = 0.56, N = 1019). We assessed reliability and responsiveness of S5 in a prospective cohort study of 41 children with sinusitis. Parents completed a questionnaire at the office visit, at 12 h and 3, 7, 10 and 14 days. Intra subject reliability at 12 h was excellent (ICC = 0.94). The S5 score was responsive in 24 patients followed for 14 days who improved (mean change = 1.52, SD = 0.12, p = 0.0062). The S5 score is standardized, reliable, responsive, easily obtained, and can be used to determine study eligibility and assess treatment effects without a physician's evaluation. PMID- 10472154 TI - A review of health-related quality-of-life concepts and measures for Parkinson's disease. AB - Parkinson's disease affects individuals health-related quality of life (HQL). Including standardized HQL assessments in therapeutic clinical trials will broaden our understanding of treatment efficacy. Selecting appropriate HQL measures for clinical studies requires consideration of their comprehensiveness, psychometric properties and feasibility. To facilitate selection, this manuscript reviews the HQL areas affected by Parkinson's disease and available Parkinson's disease-specific HQL measures: the Parkinson's Disease Questionnaire--39 (PDQ-39) and the Parkinson's Disease Quality-of-Life Questionnaire (PDQL). Based on a literature review and consultation with HQL experts, five clinicians and three patients, 12 areas of HQL were identified as particularly relevant to Parkinson's disease: physical function, mental health/emotional well being, self-image, social function, health-related distress, cognitive function, communication, sleep and rest, eating, role function, energy/fatigue, and sexual function. The PDQ-39 measures all areas except for self-image and sexual function. The PDQL measures all areas except for eating and role function. Both measures are brief and are designed and validated to be self-completed by patients. Both measures demonstrate adequate internal consistency (PDQ-39: 0.72-0.95; PDQL: 0.80-0.87) and evidence of cross-sectional validity with patient-reported measures of similar concepts. The PDQ-39 also demonstrates reproducibility (0.68-0.94), significant associations with clinical measures and preliminary evidence of responsiveness. Applications of the PDQ-39 and PDQL to clinical trials will contribute greatly towards their continued validation and interpretation. PMID- 10472155 TI - Psychometric assessment of the Life Satisfaction Questionnaire (LSQ) and a comparison of a randomised sample of Swedish women and those suffering from breast cancer. AB - In a previous study, the Life Satisfaction Questionnaire (LSQ) was developed especially for women with breast cancer. The aim of the present study was to assess the psychometric properties of the LSQ in a randomized sample of Swedish women and to compare the perceived quality of life with that of women suffering from breast cancer. Another aim was to relate the result to educational background. A total of 257 women aged 20-80, randomly selected from the Swedish population register, answered the LSQ. The result was compared with that from 362 women aged 27-78 suffering from breast cancer. The construct validity was calculated by a principal component analysis, and the reliability by Cronbach alpha-coefficients. It was concluded that the LSQ has acceptable validity and reliability. The result also showed that the women with breast cancer rated their perceived quality of life higher than women in general with respect to the quality of personal relations and the quality of daily activities, but lower for physical symptoms. In the comparison with respect to educational background, women with university education rated their quality of life higher than women with other types of education. The testing of the LSQ will continue. PMID- 10472156 TI - Quality of life in alcohol-dependent subjects--a review. AB - Although alcohol misuse is a major cause of morbidity and mortality and an important health care burden, the Quality of Life (QoL) of alcohol misusing subjects has been little studied to date. For example, only 5 out of 442 accepted abstracts at a recent international QoL conference concerned alcohol-dependent subjects. This paper reviews the ongoing and published work in the area focusing upon QoL characteristics of alcohol-dependent subjects, the link between QoL comorbidity and alcoholism, QoL alcohol dependency and social environment, changes in QoL status as a result of abstinence, minimal or controlled drinking, QoL as a predictor of relapse to heavy drinking and the importance of using a QoL measure when assessing treatment outcomes together with some of the present difficulties with existing measures. The main conclusions from the review were that the QoL of alcohol-dependent subjects is very poor but improved as a result of abstinence, controlled or minimal drinking. The important factors in the QoL of alcohol-dependent subjects are psychiatric comorbidity, social environment and disturbed sleep. PMID- 10472157 TI - Quality of life assessment in adults with type 1 Gaucher disease. AB - The effect of enzyme replacement therapy on health-related quality of life in 25 adults with type 1 Gaucher disease was investigated over a 2-year period. Quality of life was assessed using the SF-36 Health Survey (SF-36). Psychological functioning was assessed using the Symptom Checklist--90R. The results indicated significant improvement in 7 of 8 SF scale scores beginning at 18 months of therapy (P < 0.05 to 0.001). The SF scale showing improvement first was Vitality (energy level and fatigue) at 6 months of therapy (P < 0.01). The SF-36 scales showing the largest improvements were Role-Physical and Social Functioning (P < 0.001). Compared to the general US adult population, the study population's health profile was significantly lower prior to starting therapy but by 24 months of therapy there were no differences between the two. No differences were found in psychological functioning compared to a US adult normative group at the start of therapy. However, within the study population there was significant improvement in mood and global functioning and fewer psychological symptoms reported at 24 months of therapy. The findings indicate that enzyme replacement therapy for type 1 Gaucher disease has a positive impact on health-related quality of life from the patient's perspective. PMID- 10472158 TI - Do we know what global ratings of health-related quality of life measure? AB - Information is lacking about the public's perception of the term health-related quality of life (HRQL). Specifically, what are the relations between the domains included in the operational definition of HRQL tools and global health ratings. The purpose of this analysis was to identify factors associated with global rating of HRQL. We conducted a survey of a representative sample of 2,030 Israeli adults, aged 45-75 years. Multiple linear regression analysis was used to identify associations between the dependent variable, the global rating, and socio-economic details, presence of disease states, and each of the domains of the SF-36. The results demonstrate that the model explains only 52% of the variance of the global rating score. The general health domain of the SF-36 explains the vast majority of the variance, 38.5%. Another important explanatory variable was physical functioning domain, which explains 7.0% of the variance and to a lesser extent vitality. The other domains of the SF-36, socio-economic details and presence of disease states contribute only small percentages to the total explained variance of the global ratings of HRQL. It seems that there is a considerable difference between the operational definition of the research community of HRQL and the public perception of this term. PMID- 10472159 TI - Impact of weight loss on health-related quality of life. AB - To examine the effect of treatment-induced weight loss on Health-Related Quality of Life (HRQL), 38 mildly-to-moderately overweight persons recruited to participate in a study to examine the efficacy of a lifestyle modification treatment program completed a sociodemographic questionnaire, the Beck Depression Inventory (BDI), the Medical Outcomes Study Short-Form Health Survey (SF-36, as an assessment of HRQL), and underwent a series of clinical evaluations prior to treatment. After baseline evaluations, participants were randomly assigned to either a program of lifestyle physical activity or a program of traditional aerobic activity. Participants again completed the SF-36 and BDI after the 13 week treatment program had ended. Weight loss averaged 8.6 +/- 2.8 kg over the 13 week study. We found that weight loss was associated with significantly higher scores (enhanced HRQL), relative to baseline, on the physical functioning, role physical, general health, vitality and mental health domains of the SF-36. The largest improvements were with respect to the vitality, general health perception and role-physical domains. There were no significant differences between the lifestyle and aerobic activity groups on any of the study measures. These data indicate that, at least in the short-term, weight loss appears to profoundly enhance HRQL. PMID- 10472160 TI - Quality of life and depression: symmetry concepts. AB - From the theory of Three-Time-dimensions based on the disturbed temporal horizon of the depressed individual: Past, Present, therapeutic-Future, new concepts of phenomenological-depression and the intrinsic-quality of life have been defined, in which both are considered as the perception of a discrepancy between, on one hand, the healthy Past and Present ill-being, and on the other, the intensity of demand for recovery or a therapeutic-Future. In order to determine the symmetry of these two concepts, the Three-Time-dimensions Synoptic Scale (3TSS) was presented to 110 subjects who had been diagnosed as suffering from major primary depression. The demonstration was carried out in three stages, the first statistical, the second mathematical, and finally, a geometrical one; it showed the importance of the temporal horizon of the depressed person in diagnosis. PMID- 10472161 TI - Quality of life of stroke survivors. AB - Adaptation to stroke requires complex, long-term change in stroke survivors' lives. This study aimed at identifying those factors that influence quality of life (QOL) of geriatric stroke survivors 1-3 years post-discharge. The objectives were: to describe the overall quality of life of stroke survivors; to examine the relationships between sociodemographic variables, neurological variables, functional status, social support, perceived health status, depression, and overall QOL; and to determine the best predictors of QOL. Data were collected on 50 stroke survivors using a cross-sectional design and standardized questionnaires, including the Quality of Life Index, the Functional Independence Measure, the Social Support Inventory for Stroke Survivors and the Centre for Epidemiologic Studies Depression Scale. The overall quality of life of the study participants was low. The most important predictors of QOL were depression, marital status, quality of social support, and functional status. Depression was the strongest predictor of QOL. By employing a multi-dimensional perspective, this study confirmed that adaptation to stroke involves much more than physical function. Thus, rehabilitation programs for this group would be more effective if they are based upon a holistic approach. PMID- 10472162 TI - Feasibility, validity and test-retest reliability of scaling methods for health states: the visual analogue scale and the time trade-off. AB - The feasibility, validity and reliability of the Time Trade-Off (TTO) and Visual Analogue Scale (VAS) methods in obtaining preference values for health states were compared in a random sample of the Spanish population (n = 294). Respondents valued 43 EuroQol-5D health states in face-to-face interviews. Convergent validity was assessed by examining the relationship between values, and the effect of sociodemographic and health variables on values was used as a means of assessing construct validity. Test-retest reliability was analysed in a subgroup of 50 respondents, using the intraclass correlation coefficient (ICC) and generalisability theory. Rates of non-response and missing data were low on both methods, though the VAS took considerably less time to administer. VAS and TTO values correlated highly (r = 0.92), though there were differences in the ordering of health states between methods, and in the number of health states rated worse than death. VAS values were compressed into a considerably smaller valuation space than TTO values. Respondents in higher educational categories assigned higher TTO values to 12 health states. Mean ICCs (95% CI) at individual level were 0.90 (0.88-0.92) and 0.84 (0.81-0.87) for the VAS and TTO, respectively. Generalisability analysis showed variance due to time to be 0 for both methods. In conclusion, the VAS was more feasible and slightly more reliable than the TTO, whilst doubt can be cast on the degree of convergent validity existing between the two methods. The compression of VAS values means that the TTO is likely to discriminate better between health states, and it may have greater construct validity if results from larger samples confirm that there are genuine differences between sociodemographic subgroups. PMID- 10472163 TI - Factor analysis of the psychosocial items of the EORTC QLQ-C30 in metastatic breast cancer patients participating in a psychosocial intervention study. AB - The underlying factor structure of a subset of 12 items, which comprise the psychosocial subscales of the EORTC QLQ-C30 was explored in a group of women, all with metastatic breast cancer who were participating in a psychosocial intervention study. Two main factors were identified in this exploratory factor analysis, representing "emotional distress" and "functional ability" dimensions. A preliminary assessment of the external validity of the two factor structure was undertaken. The results support the validity of a summative "emotional distress" and "functional ability" score in this sample of patients. The "functional ability" score discriminated well for subgroups defined by clinical status indicators (e.g., performance status, pain, chemotherapy treatment, fatigue). The "emotional distress" subscale discriminated with respect to suffering, fatigue and sleep disturbance. Both subscales converged with related concepts measured by independent instruments, providing support for convergent validity. Summative index scores may be advantageous for application in particular research situations; applying quality adjustments in health policy analyses; for screening purposes; to monitor populations and make comparisons across broad groups and as stratification variables in clinical trials. Further research to confirm the 2 factor structure is required in other samples before the interpretation can be accepted with confidence. PMID- 10472164 TI - The relationship between demographic and clinical variables, and quality of life aspects in patients with psoriasis. AB - There is a strong need for health care programmes to promote functioning and quality of life in patients suffering from psoriasis. The aim of the present study is to highlight the relationships between demographic and clinical variables and disease-specific disability, health status, the perception of living with a chronic disease and the overall quality of life in patients suffering from psoriasis. A further aim is to examine the extent to which the effects of demographic and clinical variables on mental health and the overall quality of life are mediated by disability, physical health and the perception of living with psoriasis. The sample examined in this study comprised 334 patients (20% in-patients and 80% out-patients) who were treated consecutively at three dermatology departments in eastern Norway. A total of 282 patients completed the testing procedures, yielding a response rate of 85%. The following questionnaires were used: The Psoriasis Disability Index, the Sf-36 and the Quality of Life Scale. Correlation and multiple linear regression analyses were performed to address the core issue. Results show that demographic and clinical variables combine to explain variance in health status, the perception of living with psoriasis and overall quality of life. While most of the variance is explained by the clinical variables, the disease-specific disability variable seems to be an important mediating factor. PMID- 10472165 TI - Measuring health-related quality of life in patients with venous leg ulcers. AB - INTRODUCTION: The effect on quality of life by healing leg ulcers is not known and no validated disease-specific tool is available for measuring health-related quality of life (HRQoL) for people with venous leg ulcers. The objective of this paper was to compare four generic instruments [MOS 36-Item Short-Form Health Survey (SF-36); EuroQol (EQ); McGill Short Form Pain Questionnaire (SF-MPQ) and the Frenchay Activities Index (FAI)] used for measuring HRQoL in people with venous leg ulcers, and to offer guidance on the most appropriate tool for researchers. METHODS: Two hundred and thirty-three patients with venous leg ulcers were recruited as part of a randomised controlled trial of the cost effectiveness of community leg ulcer clinics. Subjects completed questionnaires containing the four instruments on three occasions (initial assessment, 3 and 12 months). The discriminative and evaluative properties of the four instruments were compared. RESULTS: All four instruments were acceptable to patients, taking a mean of 19.3 (SD 6.3) min to complete. At initial assessment, the SF-MPQ had poorer discriminative properties than the other three instruments and was not able to distinguish between the different patient groups in relation to age and ulcer duration. The FAI was good at discriminating between the different patient groups (at initial assessment) in relation to age, mobility and ulcer size. At the three-month follow-up, the SF-MPQ was more responsive than the other measures and detected changes in HRQoL, whereas the EQ and SF-36 did not. At 12 months, the SF-MPQ still identified differences and the SF-36 and EQ also did at this stage. CONCLUSION: In the absence of a validated condition-specific tool for measuring changes in general health status for patients with venous leg ulcers, we make the following recommendations. For evaluating the outcome of interventions with a short-term follow-up (three months) in a clinical study we recommend the SF-MPQ and for 12-month follow-up in a clinical study the SF-36, with or without the SF-MPQ. PMID- 10472166 TI - A health related quality of life measure for use in patients with urge urinary incontinence: a validation study. AB - OBJECTIVE: Urge incontinence has substantial impact on health related quality of life (HRQOL). The purpose of this research was to test the psychometric properties of the Urge-Incontinence Impact Questionnaire (U-IIQ) and Urge-Urinary Distress Inventory (U-UDI). METHODS: The U-IIQ and the U-UDI were incorporated in a single arm study of an investigational once-a-day formulation of oxybutynin (Ditropan XL) for urge incontinence. The U-IIQ contains 7 scales scored separately: Travel, Activities, Physical activities, Feelings, Relationships, Sexual function and Nighttime bladder control. We also calculated a single index score. The U-UDI contains 9 items summarized in a single score that measures the extent to which incontinence symptoms bother patients and an urge symptoms summary score. Patients completed the questions at: start of baseline week, end of baseline week, week 4 and week 12 (final dose). RESULTS: The questions were completed by the 257 patients enrolled in the study (91% female, mean age = 60 years). All item responses showed good variability. Internal-consistency reliability was very good (Cronbach's alpha = 0.82-0.96). Test-retest reliability was good for all scales ([ICC] = 0.68-0.83). Discriminant validity was noted for all scales for patients with and without frequent incontinence (p < or = 0.006 all scales). All scales demonstrated responsiveness to change over time (Guyatt's statistic = magnitude of -1.04 - magnitude of -1.71). CONCLUSIONS: Psychometric testing indicated that the scales measured distinct and relevant domains of HRQOL for patients with urge incontinence, and that these two instruments are reliable, valid, and responsive to change in this condition. PMID- 10472167 TI - Quality of life and Parkinson's disease: translation and validation of the US Parkinson's Disease Questionnaire (PDQ-39). AB - Parkinson's disease is a common progressive neurodegenerative disorder affecting an estimated 4 million people worldwide. A number of general health status measures exist but few fully capture the subjective evaluation of life quality associated with Parkinson's disease. We report here the results of: (1) translating the British PDQ-39 into a US version, (2) validity and reliability of the new US PDQ-39 questionnaire, and (3) parallel validation analyses following the method published in the development of the British version of the PDQ-39. Data were collected by postal survey on 150 patients recruited from neurology clinics in the Seattle area. A short, generic health status measure (SF-36) was used to test convergent validity, and a three-day test-retest assessed the reliability of the PDQ-39. The US version of the PDQ-39 demonstrated acceptable internal consistency (alpha = 0.51 to 0.96) and proved to be reproducible (0.86 to 0.96). Subscales of the PDQ-39 showed convergence with like scales of the SF 36 and was able to discriminate between levels of symptom severity. PMID- 10472168 TI - The SF-36 as a health status measure for epilepsy: a psychometric assessment. AB - A considerable literature is now available on the applications and psychometric properties of the MOS SF-36 Health Survey. In epilepsy, the SF-36 has been used as a health status measure in its own right and as the stem for two condition specific measures. This paper replicates for epilepsy previous work to support use of the SF-36 across a range of clinical conditions. Data were obtained from a European-wide descriptive study of quality of life of adults with epilepsy; analysis of responses on the SF-36 is based on 4,929 subjects in eight countries. Missing value rates for all SF-36 items were low; percentages for whom complete information available for subscales ranged from 95.7% to 98.6%. All subscales passed tests for item-internal consistency and item-discriminant validity. Reliability coefficients exceeded the standard recommended for group comparisons across all subscales. Floor effects were negligible for all but the two role disability subscales; there were substantial ceiling effects for five of the SF 36 subscales. We conclude that the SF-36 is a valid and reliable health status measure for descriptive studies of people with epilepsy, but ceiling effects may limit its usefulness as an outcome measure in the assessment of new treatments. PMID- 10472169 TI - Recurrent genital herpes and quality of life in France. AB - Recurrent genital herpes is an incurable disease that may give rise to considerable psychological disturbance. This paper describes the impact of the disease on quality of life in a French population. The population study was also used to provide additional validation data for the French version of the RGHQoL (the Recurrent Genital Herpes Quality of Life) measure. The content of the RGHQoL was derived from patient interviews in the UK. The measure was then developed and tested simultaneously in several countries, including France. Statements in the measure were translated into French by a bilingual panel and then by a lay panel. Field-testing in France took place with 15 patients who judged the instrument to be relevant and easy to complete. The final version of the questionnaire was assessed for reliability and construct validity. The measure was found to have good test-retest reliability (0.94) and internal consistency (0.93) as assessed by Cronbach's alpha. The French RGHQoL was then used with 150 herpes patients obtained from a general population sample. A majority of the respondents completed all questions, indicating a high level of acceptability. Quality of life was found to be lowest in the under 25-year-olds and in those respondents who were most psychologically and sexually disturbed by their outbreaks and found it difficult to discuss the problem with their partner. It is concluded that the French version of the RGHQoL is an acceptable, reliable and valid instrument and that the measure provides valuable information about the impact of the disease. PMID- 10472170 TI - The QoL-AGHDA: an instrument for the assessment of quality of life in adults with growth hormone deficiency. AB - Several studies have shown that growth hormone deficiency in adults leads to poor well-being and other clinical consequences, and that these improve when the hormone is replaced. However, the studies employed generic measures of health status that miss important aspects of the patients' experience and that have inadequate reliability and responsiveness. This paper describes the European development and testing of the Quality of Life-Assessment of Growth Hormone Deficiency in Adults (QoL-AGHDA), a condition-specific quality of life measure for use in clinical trials and for the routine monitoring of patients. The instrument was produced in five languages; English, Swedish, Italian, German and Spanish. Each language version is shown to have good reliability, internal consistency and construct validity. The QoL-AGHDA is currently included in an international database monitoring the long-term efficacy and safety of growth hormone replacement therapy and in clinical trials in a number of countries. PMID- 10472171 TI - Health related quality of life and disease severity in patients with alpha-1 antitrypsin deficiency. AB - STUDY QUESTION: To describe health-related quality of life (HRQL) in individuals with alpha-1 antitrypsin (AAT) deficiency, examine the cross-sectional relationship between disease severity and HRQL, and explore changes in lung function and HRQL over time in a subset of these individuals. MATERIAL/METHODS: Forty-five adults with AAT deficiency and moderate to severe emphysema completed the Chronic Respiratory Disease Questionnaire (CRQ), six-minute walk distance (6 MWD) and pulmonary function tests (PFTs). Twenty of the 45 were followed for two additional years with repeated measurements of CRQ and PFTs. RESULTS: The mean +/ SD age was 49 +/- 8 years. Initial CRQ subscale scores were: dyspnea 17.5 +/- 4.3; fatigue 17.0 +/- 5.46; emotional function 33.1 +/- 8.67; and mastery 21.7 +/ 4.65. No relationship was found between percent predicted forced expiratory volume in one second (FEV1%) and CRQ score; 6 MWD and fatigue correlated significantly (r = 0.32, p < 0.05). Repeated PFT and CRQ measurements in 20 subjects showed statistically significant declines in FEV1 and slow vital capacity (SVC), but no change in CRQ scores. CONCLUSIONS: Results suggest persons with AAT deficiency face challenges to HRQL that are similar to older adults with chronic pulmonary disease. Further research is needed on the nature of the relationship between disease severity and HRQL in this population. PMID- 10472172 TI - Communication pathology: the way in Africa. AB - Universities in Southern Africa are currently deeply involved in transformation processes in order to ensure world class tertiary education. More specifically in the case of vocationally oriented tertiary programmes, it has become necessary to provide answers to questions like: are the academic outcomes of higher education relevant to and accountable in the context of the workplace? For the Department of Communication Pathology at the University of Pretoria the answer is unequivocal--a programme that aims to provide learning opportunities for professionals who must eventually render a service to a specific population (the individual with a speech, voice, language or hearing disorder), should ensure that the characteristics of this population serve as the basis for strategic planning. This population is an African population. Educational programmes, research projects and community service must therefore be africanised. PMID- 10472173 TI - [Afrikaanse Reseptiewe Woordeskattoets (ARW) : Suitability for a group of non standard Afrikaans speaking children]. AB - The need for culturally appropriate language tests served as motivation for this study. The aim of the study was to evaluate the suitability and adaptability of the Afrikaanse Reseptiewe Woordeskattoets (ARW) (Buitendag, 1994) for a group of seven and ten year old non-standard Afrikaans speaking children from the coloured community in Gauteng, North West, Mpumalanga and Northern Province. In order to achieve this goal the ARW (Buitendag, 1994), and a questionnaire regarding socio economic deprivation, were administered to a proportionally stratified randomized sample of 960 children. Test bias was evaluated by comparing the psychometric characteristics of the test for the research group to those of the normative group. Various test adaptation procedures were considered, namely adaptations to the rank order of item difficulty, test starting points and test procedures. The results indicated the same degree of confidence in the test results of the two groups. It demonstrated that the original rank order of item difficulty does not need to be changed and that corresponding increase in ARW scores is evident with an increase in age in both groups. The findings imply that the test measures the same ability in the two groups. Although comparison of the psychometric characteristics of the test for the research group to those for the normative group failed to identify test bias, intra-group comparison indicated that the test may be linguistically biased against the socio-economically deprived sub group. Significant differences in individual ARW scores occurred when a shift in the child's basal or ceiling item occurred as a result of the biased items. The test's cultural validity for the group may be increased by lowering the test starting points, in order to prevent reversed testing. Culturally appropriate responses should be accepted as correct, in order to award equal credit to standard and non-standard language responses and to take intra-group differences into consideration. This research is an important contribution towards the expansion of the speech-language therapist's skills and resources, vitally needed to provide culturally relevant services in South Africa to all individuals with communication disorders. PMID- 10472174 TI - Babbling of an infant with a repaired cleft lip: a case study. AB - The infant born with a cleft lip is faced with risk factors that threaten the development of speech-language skills. The earlier the age of identification and management of the developmental delay, the better the outcome. The attainment of the mature syllable is considered to be a critical measure of babbling competency. This single case study aimed to determine whether the formedness of the syllable in babbling would be affected by the cleft lip repaired prior to the onset of meaningful speech. Three samples of babbling in a naturalistic environment were video-recorded. Data was analysed following the principles of infraphonology, employing a perceptually-based method. A profile of infraphonological features was obtained. Results showed that the development of the mature syllable was attained. The results support the theories that babbling is a robust phenomenon. Clinical and research implications are discussed. PMID- 10472175 TI - Attitudes of a group of primary school teachers towards the educational inclusion of hearing-impaired learners in regular classrooms. AB - Research has clearly demonstrated a link between the attitudes of regular education teachers and the success of inclusion of learners with special educational needs. Therefore, the present study aimed to investigate the attitudes of a group of junior primary school teachers from the Gauteng area towards the inclusion of hearing-impaired children into regular classes. A survey research design was employed which utilized a questionnaire as the research tool. Analysis of results indicated that the teachers surveyed were relatively positive in their attitudes towards inclusion. Greater exposure to disability in terms of training and experience was related to more positive attitudes. Similarly, more positive attitudes were related to greater perceived competence in teaching hearing-impaired pupils. All of the teachers surveyed felt that speech-language pathologists and audiologists (SLPs & As) should be involved in facilitating inclusion of hearing-impaired children. Many of the respondents expressed concern regarding their lack of training, knowledge and skills. The findings from the research project highlight the need for an adequate training and support system for teachers prior to the implementation of an inclusive educational policy, and the potential role of SLPs & As in this regard. PMID- 10472176 TI - Early communication intervention with young children with Pierre Robin sequence. AB - Young children with Pierre Robin sequence are at considerable risk to develop delayed or disordered communication development. This study investigated the effectiveness of early communication intervention with four young children with Pierre Robin sequence, aged 5 to 28 months. The Proportional Change Index (Wolery, 1983) was used to determine the amount of child progress that took place during the intervention period. The results indicated that regular early communication intervention sessions over an extended period of time produced positive results, especially for expressive language abilities. The presence of a slight hearing loss due to otitis media, seemed to have influenced the effectiveness of early communication intervention. One subject displayed an associated disorder and evidenced the slowest rate of development for receptive and expressive language abilities as well as a limited phonetic repertoire for consonants. All but one subject had limited phonetic repertoires possibly due to velopharyngeal incompetence. Early communication intervention services should be delivered regularly over an extended period of time and the hearing abilities and speech production skills of young children with Pierre Robin sequence should be followed closely during early communication intervention. PMID- 10472177 TI - Perceptions of tinnitus in a group of senior citizens. AB - Although previous research has highlighted the debilitating effects of tinnitus on quality of life, most studies appear to have targeted the general population, with few researchers focusing specifically on elderly persons. Hence, the purpose of this study was to investigate the perceptions of tinnitus in a group of 20 senior citizens. A questionnaire based on existing research instruments developed by Stouffer and Tyler (1990) and Wilson et al. (1991) was used to elicit information regarding the auditory-perceptual characteristics of tinnitus, effects of tinnitus on lifestyle, perceived causes, factors aggravating or ameliorating tinnitus and coping strategies employed. Results supported findings from earlier research and indicated that tinnitus did indeed exert an impact on respondents' lives with 65% avoiding noisy situations, 65% having difficulty relaxing, and 55% struggling to follow conversations. Almost two-thirds, namely 70%, of respondents reported feelings of frustration, 60% and 55% mentioned feelings of tenseness and irritability respectively, while 50% experienced depression associated with the tinnitus. These findings are discussed in terms of their implications for the education of health care professionals, management of tinnitus patients in general, public and patient awareness campaigns, the establishment of Tinnitus Self Help Groups, and further research. PMID- 10472178 TI - Use of the NAL-AB wordlists as a South African English speech discrimination test. AB - South Africa still lacks a South African English specific speech discrimination test. As an alternative, this study investigated the use of the Australian English, National Acoustic Laboratories Arthur Boothroyd (NAL-AB) wordlists to assess the speech discrimination of South African English speakers. Thirty South African English speakers were tested at 0, 5, 10, 20 and 25 dBHL (audiometer dial reading) and their performance-intensity functions were compared qualitatively to the NAL-AB wordlist normative data. Results showed three general patterns; similar performance for both groups; poorer performance by the South African English speakers at the low to mid presentation intensities only; and poorer performance by the South African English speakers across most presentation intensities. Use of the NAL-AB wordlists at threshold levels or for site of lesion assessment was therefore concluded to be unwise. Use of these wordlists at supra-threshold levels, however, would provide a valid and reliable option for the speech discrimination assessment of South African English speakers. PMID- 10472179 TI - An alternative non-traditional approach to learning: the metacognitive-mapping approach. AB - The current article is a continuation of a previous article (Leaf, Uys & Louw 1997) suggesting that alternatives to the current traditional perceptions of learning and therapeutic methods are essential if learning institutions are to enable learners to move successfully into the next millenium. The need for the speech-language therapist to adopt a non-traditional approach to learning and to change from a language instructor to a learning facilitator within a consultative collaborative role in the unique South African situation is discussed. Research results, specifically relating to how the speech-language therapist can assist teachers in changing to a non-traditional facilitative role, are presented. PMID- 10472180 TI - The ASHA way: growth through standards. American Speech-Language Hearing Association. PMID- 10472181 TI - The function of vestigial in Drosophila wing development: how are tissue-specific responses to signalling pathways specified? AB - The activities of conserved signal transduction pathways are central to the development of Drosophila wings, legs, and eyes. Yet, all these structures have characteristic morphologies, suggesting that additional factors provide organ specific information. One excellent candidate for such a function is Vestigial, which activity promotes the formation of wings. The biochemical function of Vestigial is unknown, however, since no homologies with other proteins have been identified. Two recent reports show that Vestigial interacts with the transcription factor Scalloped, forming an active complex that binds to specific DNA sequences and regulates gene expression in cooperation with several signalling pathways. These results illustrate how tissue-specific transcription factors cooperate with general signalling pathways to regulate gene expression in a tissue-specific manner. PMID- 10472182 TI - Polarizing genetic information in the egg: RNA localization in the frog oocyte. AB - RNA localization is a powerful strategy used by cells to localize proteins to subcellular domains and to control protein synthesis regionally. In germ cells, RNA targeting has profound implications for development, setting up polarities in genetic information that drive cell fate during embryogenesis. The frog oocyte offers a useful system for studying the mechanism of RNA localization. Here, we discuss critically the process of RNA localization during frog oogenesis. Three major pathways have been identified that are temporally and spatially separated in oogenesis. Each pathway uses a different mechanism to effect RNA localization. In some cases, localization elements within the 3' untranslated region have been identified and have provided unique insights into the localization process. This important field is still in its infancy, however, and much remains to be learned. PMID- 10472183 TI - Recognition of sorting signals by clathrin adaptors. AB - Sorting of membrane proteins is generally mediated by cytosolic coats, which create a scaffold to form coated buds and vesicles and to selectively concentrate cargo by interacting with cytosolic signals. The classical paradigm is the interaction between clathrin coats and associated adaptor proteins, which cluster receptors with characteristic tyrosine and dileucine motifs during endocytosis. Clathrin in association with different sets of adaptors is found in addition at the trans-Golgi network and endosomes. Sequences similar to internalization signals also direct lysosomal and basolateral sorting, which implicates related clathrinadaptor coats in the respective sorting pathways. This review concentrates on the recognition of sorting signals by clathrin-associated adaptor proteins, an area of significant recent progress due to new methodological and conceptual approaches. PMID- 10472184 TI - DEG/ENaC channels: a touchy superfamily that watches its salt. AB - To the surprise of many, studies of molecular mechanisms of touch transduction and analyses of epithelial Na+ transport have converged to define a new class of ion channel subunits. Based on the names of the first two identified subfamilies, the Caenorhabditis elegans degenerins and the vertebrate epithelial amiloride sensitive Na+ channel, this ion channel class is called the DEG/ENaC superfamily. Members of the DEG/ENaC superfamily have been found in nematodes, flies, snails, and vertebrates. Family members share common topology, such that they span the membrane twice and have intracellular N- and C-termini; a large extracellular loop includes a conserved cysteine-rich region. DEG/ENaC channels have been implicated a broad spectrum of cellular functions, including mechanosensation, proprioception, pain sensation, gametogenesis, and epithelial Na+ transport. These channels exhibit diverse gating properties, ranging from near constitutive opening to rapid inactivation. We discuss working understanding of DEG/ENaC functions, channel properties, structure/activity correlations and possible evolutionary relationship to other channel classes. PMID- 10472185 TI - Fibrils as extracellular appendages of bacteria: their role in contact-mediated cell-cell interactions in Myxococcus xanthus. AB - Social behavior in the myxobacterium Myxococcus xanthus involves epicellular, peritrichous appendages called fibrils. These are polysaccharide organelles containing a set of tightly adhering proteins. It is proposed that cell-cell contact is perceived by the fibrils and is mediated by the action of a fibrillar ADP-ribosyl transferase. Fibrils or fibril-like organelles have also been found on a variety of other gram-negative bacteria and at least one archaeon, and may mediate cell-cell contact between the bacteria themselves or between the bacteria and their eukaryotic host cells. PMID- 10472186 TI - Accessing DNA damage in chromatin: insights from transcription. AB - Recently, there has been a convergence of fields studying the processing of DNA, such as transcription, replication, and repair. This convergence has been centered around the packaging of DNA in chromatin. Chromatin structure affects all aspects of DNA processing because it modulates access of proteins to DNA. Therefore, a central theme has become the mechanism(s) for accessing DNA in chromatin. It seems likely that mechanisms involved in one of these processes may also be used in others. For example, the discovery of transcriptional coactivators with histone acetyltransferase activity and chromatin remodeling complexes has provided possible mechanisms required for efficient repair of DNA in chromatin. PMID- 10472187 TI - Vertebrate evolution: the developmental origins of adult variation. AB - Many biologists assume, as Darwin did, that natural selection acts mainly on late embryonic or postnatal development. This view is consistent with von Baer's observations of morphological divergence at late stages. It is also suggested by the conserved morphology and common molecular genetic mechanisms of pattern formation seen in embryos. I argue here, however, that differences in adult morphology may be generated at a variety of stages. Natural selection may have a major action on developmental mechanisms during the organogenetic period, because this is when many adult traits are specified. Evolutionary changes in these early developmental mechanisms probably include subtle shifts in the timing of gene expression. Changes of this kind have little or no gross effect on the anatomy of the embryo; they are only phenotypically expressed, or readily detected, when amplified at later stages. The phylotypic stage, the developmental hourglass, modularity, and von Baerian divergence are reassessed in terms of these arguments. PMID- 10472188 TI - Declining semen quality: can the past inform the present? AB - By using instrumentation initially designed for counting white blood cells, sperm counts have been utilized by clinicians since 1929, particularly to evaluate cases of suspected infertility. Although this basic biological parameter might be assumed to be stable over time, several studies over the past 20 years have suggested a decline in sperm count or density. The most controversial of these analyses was published in 1992. A flood of criticism followed this analysis of 61 studies that found a 50% decline in sperm density between 1938 and 1990. Critics suggested that historical methods (of counting sperm or conducting studies) were variable and unreliable, differing from modern methods both qualitatively and quantitatively. To address this issue we analyzed these studies for trends in counting methods or their variability. We found neither. Alternative analyses produced some differences in trend estimates, but statistical factors alone could not account for the total decline in sperm density. We reviewed study populations to identify trends in population characteristics, such as abstinence time, that might explain the decline. However, controlling analytically for such factors only increased the rate of decline. We conclude that historical data on sperm density, despite large random error, are surprisingly reliable. Nonetheless, understanding causes of temporal and geographic differences in sperm density must await contemporary data. PMID- 10472189 TI - Simulants, stimulants and diseases: the evolution of the United States biological warfare programme, 1945-60. AB - Details about the US biological programme have largely been based on information in the open literature. More revealing aspects of the programme are now available through documents released under the Freedom of Information Act. Annual reports of the activities of the US Army Chemical Corps from 1945 to 1959 have revealed significant increases in activity in biological warfare research. The Corps research activity progressed from work on anthrax in 1941, through anti-crop agents in the mid-1940s, to a wider range of animal, plant and human diseases by 1960. A number of disease organisms were investigated sufficiently to permit some standardisation and manufacture of munitions. PMID- 10472190 TI - A magic sword or a big itch: an historical look at the United States biological weapons programme. AB - In the late 1950s interest in entomological warfare increased, and literature describing the US biological warfare programmes on the use of the mosquito Aedes aegypti, the vector for transmitting yellow fever, has now been released. Yellow fever was considered as a suitable disease to use in southern regions of the former Soviet Union. The US destroyed its biological weapon stockpiles in the early 1970s. In addition to its offensive biological warfare programme, the US conducted extensive trials to assess its own vulnerability to biological attack. These trials and a later series of threat analyses indicate that biological agents could, indeed, affect large areas of the US if the attackers were allowed to proceed unmolested. Some of the threat analyses present highly questionable scenarios. PMID- 10472191 TI - International relations and epidemics: a short expedition to places inhabited by states and mad cows. AB - The complex process of co-evolution between humans, their social structures and biological disease agents have from time to time established relationships between the three. Recently, one such set of paths has opened up faster and closer global connections. As new and more inclusive approaches emerge from the shadow of strict intra-disciplinary containment, it is tempting to formulate the relationship between epidemics and international relations in new terms deriving from new metaphors. The argument that components central to international relations (state, sovereignty, power) do not associate with disease can be viewed sceptically. Throughout history, epidemics have had a direct impact on political interaction by vindicating, weakening, testing, and moulding international relations. This paper examines the past relationships between epidemics and international relations, and illustrates these interactions with the example of Bovine Spongiform Encephalopathy (mad cow disease) and its link with human Creutzfeldt-Jakob disease. PMID- 10472192 TI - Recent medical evidence for torture and human rights abuse in Sierra Leone: a report for the Medical Foundation for the Care of Victims of Torture. AB - A study was carried out using the case notes of all Sierra Leoneans referred to the Medical Foundation for the Care of Victims of Torture between 1996 and 1998 for medical reports. Data were abstracted on the type and extent of torture reported, and the cases were followed up to the present day to ascertain how long the United Kingdom asylum and immigration process had taken to process their claims. In the sample of 36 cases there were 68 reported incidents of torture or human rights abuse. Sixty-nine per cent (18/26) of the women in the sample had been raped at least once, and 72% (26/36) of the sample had been beaten at least once. The average time for the minimum due process of the asylum claim to be processed was 44 months (3.7 years). On 31 January 1999, 61% of the sample (22 people) were still involved in the asylum process. Waiting times must be reduced and safeguards imposed so that those who have suffered torture can be rapidly identified and referred for medical treatment, care and documentation. PMID- 10472193 TI - The BMA report on biological and genetic weapons. PMID- 10472195 TI - Vision and visual cognition. PMID- 10472194 TI - David Pimentel on the ecology of increasing disease: population growth and environmental degradation. PMID- 10472196 TI - The psychophysics of perceptual memory. AB - Psychophysical studies of short-term memory for attributes or dimensions of the visual stimulus known to be important in early visual processing--spatial frequency, orientation, contrast, motion--identify an early perceptual memory system. The proposed system, which may be part of the Schacter-Tulving perceptual representation system (PRS), is located early in the visual processing stream, prior to the structural description system responsible for shape priming but beyond primary visual cortex (V1), and consists of a series of parallel special purpose perceptual mechanisms with independent but limited processing resources, where each mechanism is devoted to the analysis of a single stimulus dimension and is coupled to a memory store. The experimental evidence for this hypothesis is reviewed. PMID- 10472197 TI - Visual and mental exploration of visuo-spatial configurations: behavioral and neuroimaging approaches. AB - Do mental imagery and perception involve common processing mechanisms? Imagery researchers have devoted a great deal of effort to establishing the functional and structural similarities between images and perceptual events. Recent studies have focused on the comparison of images that are reconstructions of previous perceptual experience and images constructed from verbal descriptions. This article reports the findings of a research program based on the mental scanning paradigm; they reveal the similarities and differences between the two kinds of mental images. Neuroimaging studies have also provided evidence that the parieto occipital cortex is involved in the processing of visual images, whether they are based on perceptual experience or constructed from linguistic inputs. However, the PET studies conducted by our research groups provide no evidence that the primary visual cortex is engaged in the generation of visual images. As there is contradictory evidence about this, further research is needed to clarify the role of the early visual areas in mental visual imagery. PMID- 10472198 TI - From objects to names: a cognitive neuroscience approach. AB - To name an object, we need both to recognize it and to access the associated phonological form, and phonological retrieval itself may be constrained by aspects of the visual recognition process. This paper reviews evidence for such constraints, drawing on data from experimental psychology, neuropsychology, functional imaging, and computational modelling. Data on picture identification in normal observers demonstrate that the speed of name retrieval processes differs for natural objects and artifacts, due at least in part to differences in visual similarity between exemplars within these categories. Also, effects of variables on early and late stages of object identification combine in an interactive rather than an additive manner, consistent with object processing stages operating in a continuous rather than a discrete manner. Neuropsychological evidence supports this proposal, demonstrating that subtle perceptual deficits can produce naming problems, even when there is good access to associated semantic knowledge. Functional activation studies further show increased activity in visual processing areas when conditions stress object naming relative to the recognition of familiar object structures. These studies indicate that object naming is based on a series of continuous processing stages and that naming involves increased visual processing relative to recognition tasks. The data can be modelled within an interactive activation and competition framework. PMID- 10472199 TI - Subordinate-level object classification reexamined. AB - The classification of a table as round rather than square, a car as a Mazda rather than a Ford, a drill bit as 3/8-inch rather than 1/4-inch, and a face as Tom have all been regarded as a single process termed "subordinate classification." Despite the common label, the considerable heterogeneity of the perceptual processing required to achieve such classifications requires, minimally, a more detailed taxonomy. Perceptual information relevant to subordinate-level shape classifications can be presumed to vary on continua of (a) the type of distinctive information that is present, nonaccidental or metric, (b) the size of the relevant contours or surfaces, and (c) the similarity of the to-be-discriminated features, such as whether a straight contour has to be distinguished from a contour of low curvature versus high curvature. We consider three, relatively pure cases. Case 1 subordinates may be distinguished by a representation, a geon structural description (GSD), specifying a nonaccidental characterization of an object's large parts and the relations among these parts, such as a round table versus a square table. Case 2 subordinates are also distinguished by GSDs, except that the distinctive GSDs are present at a small scale in a complex object so the location and mapping of the GSDs are contingent on an initial basic-level classification, such as when we use a logo to distinguish various makes of cars. Expertise for Cases 1 and 2 can be easily achieved through specification, often verbal, of the GSDs. Case 3 subordinates, which have furnished much of the grist for theorizing with "view-based" template models, require fine metric discriminations. Cases 1 and 2 account for the overwhelming majority of shape-based basic- and subordinate-level object classifications that people can and do make in their everyday lives. These classifications are typically made quickly, accurately, and with only modest costs of viewpoint changes. Whereas the activation of an array of multiscale, multiorientation filters, presumed to be at the initial stage of all shape processing, may suffice for determining the similarity of the representations mediating recognition among Case 3 subordinate stimuli (and faces), Cases 1 and 2 require that the output of these filters be mapped to classifiers that make explicit the nonaccidental properties, parts, and relations specified by the GSDs. PMID- 10472200 TI - Serial attention mechanisms in visual search: a critical look at the evidence. AB - Until a few years ago, visual search tasks were of exclusive pertinence to psychophysicists and cognitive psychologists trying to understand the operating principles and computational constraints of visual perception and visual selective attention. In recent years, cognitive neuroscience, with its powerful tools, has started to explore more directly the neuronal mechanisms underlying search performance in humans and macaques, aiming at the same general goals. New observations from a number of cognitive neuroscience approaches are promising a near future of great excitement in this field of research. This article offers a critical review of some of these recent contributions and highlights some of the interpretational problems that they pose. PMID- 10472201 TI - Visual-spatial working memory, attention, and scene representation: a neuro cognitive theory. AB - This paper addresses the issue of how visual-spatial working memory, attention, and scene representation are related. The first section introduces a modified two stage conception of visual-spatial processing. "Stage one" refers to low-level visual-spatial processing and computes in parallel for the currently available retinal information "object candidates," here called "visual-spatial units." An attentional process called "unit selection" allows access to stage two for one of these units at a time. Stage two contains high-level visual-spatial information that can be used for goal-directions (e.g., verbal report, grasping). It consists of three parallel processing streams. First, the currently selected unit is recognized; second, a spatial-motor program for the selected unit is computed; and third, an "object file" is set up for the selected unit. An object file contains temporary episodic representations of detailed high-level visual-spatial attributes of an "object" plus an "index." An index acts as a pointer and is bound via temporary connections to the attributes of the file. Section two of this paper specifies one part of stage two in more detail, namely visual-spatial working memory (VSWM). It can contain up to four object files. A first central claim is that during sensory-based processing for working memory ("access"), one object file is always "on-line," and up to three other object files are "off line". A second central claim is that the process of setting up an object file depends on the number and the activation level of already stored files. Based on the concept of activation-based competition between object files, it is postulated that the more files that are stored and the higher their activation is, the longer it takes for a newly set up object file to reach a sufficient level of activation. Activation-based competition is also used to explain "short term forgetting" by "interference." A third central claim about VSWM is that a "refreshment" process exists that increases the activation level of an index of an object file in order to prevent forgetting or in order to bring the file back to the state of controlling the current action. Finally, section three gives a selective look at a number of experimental data such as the attentional blink, backward masking, dwell time effects, transsaccadic memory, and change blindness. New explanations are offered and new predictions made. PMID- 10472203 TI - A road map for staying safe behind the wheel. PMID- 10472202 TI - Seeds of hope, seeds of doubt. PMID- 10472204 TI - Effective medical tests you can do at home. PMID- 10472205 TI - Can infections cause heart attacks? PMID- 10472206 TI - Menopause retrospective. PMID- 10472207 TI - You recently wrote that cholesterol-lowering drugs should be taken in the evening -except for Lipitor. If that's the case, when should I take Lipitor? PMID- 10472208 TI - Beta-blockers and anticonvulsants have not helped my essential tremor. I've heard botulinum toxin might work. Is it worth trying? PMID- 10472209 TI - Rapacuronium (Org 9487): do we have a replacement for succinylcholine? PMID- 10472210 TI - Tramadol revisited. PMID- 10472211 TI - Stability of the interstitial matrix after crystalloid fluid loading studied by volume kinetic analysis. AB - To investigate if fluid therapy changes the prerequisites for the development of oedema, four i.v. infusions of Ringer's solution 25 ml kg-1 were given over 15 or 30 min in a randomized crossover study to 10 healthy male volunteers, aged 28-40 (mean 31) yr. Blood haemoglobin concentration, measured every 5 min for 90 min, and urinary excretion were used as input data for volume kinetic analysis. The results showed that the elimination rate constant (kr) was higher when another infusion had been given earlier on the same day (208 vs 140 ml min-1; P < 0.002) and the size of V1 was larger during the 15-min infusions (4.7 vs 3.2 litre; P < 0.02). However, the size of V2 and the rate constant for the exchange of fluid between V1 and V2 were similar during all infusions. We conclude that a fluid challenge makes elimination of further infused fluid more effective but does not change compliance with volume expansion in healthy volunteers. PMID- 10472212 TI - Consequences of acute normovolaemic haemodilution on haemostasis during major orthopaedic surgery. AB - Acute preoperative normovolaemic haemodilution (NHD) is an accepted tool for reducing allogeneic blood transfusion requirements during surgery. At present, little is known of its impact on haemostasis. We have investigated the consequences of NHD on haemostasis by comparing conventional global tests (prothrombin time (PT), activated partial thromboplastin time (aPTT) with more specific measures of coagulation (prothrombin fragment 1 + 2 (F 1 + 2), thrombin antithrombin III complex (TAT) and fibrinolysis (D-dimer (DD), plasmin-alpha 2 antiplasmin complex (PAP)). Blood samples were collected from two groups (NHD and controls) undergoing elective spinal surgery or pelvic osteotomy until day 3 after operation. The conventional global tests remained within normal limits: there were no significant differences between groups. Although surgery induced significant increases in the more specific measures of coagulation and fibrinolysis, there were no differences between NHD and control patients. Major orthopaedic surgery strongly activates coagulation and fibrinolysis. As the degree of these alterations was similar in haemodiluted and control patients, we suggest that acute preoperative normovolaemic haemodilution itself does not appear to be associated with greater perioperative disturbances in haemostasis. PMID- 10472213 TI - Tissue deposits of hydroxyethyl starch (HES): dose-dependent and time-related. AB - Tissue deposits occur after administration of plasma substitutes. After hydroxyethyl starch (HES), deposits may last for months, causing pruritus and impairment of function. Because elimination of HES deposits has not been demonstrated in humans, we studied 26 patients, for up to 7 yr after HES administration, to assess HES storage. HES dose ranged from 0.34 to 15.00 g kg-1 body weight, and administration intervals from 1 day to 7 yr. Biopsies of the liver, muscle, spleen, intestine or skin were studied using light and electron microscopy and immunohistochemistry. HES storage was dose-dependent, decreased in all organs with time and was greater in patients suffering from pruritus. We conclude that tissue deposition of HES is transitory and dose-dependent, with differences between subjects in severity and duration. PMID- 10472214 TI - Haemodynamic effects of propofol vs thiopental in infants: an echocardiographic study. AB - Rapid i.v. induction of general anaesthesia is indicated in infants at risk of vomiting or regurgitation to reduce the risk of aspiration of gastric contents. Propofol is an alternative to thiopental in infants, and we have compared cardiovascular changes when propofol or thiopental was used for induction of anaesthesia in infants. Twenty infants, ASA I or II, aged 1-11 months, undergoing elective surgery were allocated randomly to receive either thiopental or propofol for i.v. induction. Cardiovascular and echocardiographic data were recorded in both groups before, during and for 5 min after induction of anaesthesia. Doses required to induce anaesthesia in each group were mean 10.3 (SD 0.9) mg kg-1 of thiopental and 6.1 (0.6) mg kg-1 of propofol. Thiopental did not alter significantly systolic or mean arterial pressure, afterload indices, rate corrected velocity of circumferential fibre shortening or cardiac index, but decreased shortening fraction at 1 and 5 min after induction compared with awake values. Propofol did not alter heart rate, shortening fraction, rate-corrected velocity of circumferential fibre shortening or cardiac index at 1 and 5 min after i.v. induction compared with awake values. After induction, systolic and mean arterial pressures and afterload indices decreased more after induction with both agents, but did not become abnormal. Thus propofol decreased arterial pressure more than thiopental because of an effect on afterload. Cardiac output remained unchanged with both agents. PMID- 10472215 TI - Continuous monitoring of jugular bulb oxyhaemoglobin saturation using the Edslab dual lumen oximetry catheter during and after cardiac surgery. AB - Fibreoptic jugular bulb oximetry has been validated for use in the care of severely head-injured patients. We compared bench and fibreoptic methods of measuring jugular bulb oxyhaemoglobin saturation (SjO2) in 33 patients undergoing cardiac surgery both during cardiopulmonary bypass (CPB) and in the early postoperative period. After insertion of a fibreoptic reflectance oximetry catheter into the jugular bulb, it was calibrated against a bench oximeter. Comparisons were made while on CPB (n = 60) and in the postoperative period for up to 18 h (n = 215). There was negligible bias throughout. There were wide limits of agreements (mean difference +/- 2SD) between the two methods during operation (-20.29% to 18.05%), whereas after operation the limits of agreement were far narrower (-6.39% and 7.45%). Measurement of SjO2 by the fibreoptic method compared poorly with bench oximetry during CPB but there was good agreement between the two methods in the early postoperative period. PMID- 10472216 TI - Continuous cardiac output measurement: pulse contour analysis vs thermodilution technique in cardiac surgical patients. AB - We have analysed the clinical agreement between two methods of continuous cardiac output measurement pulse contour analysis (PCCO) and a continuous thermodilution technique (CCO), were both compared with the intermittent bolus thermodilution technique (BCO). Measurements were performed in 26 cardiac surgical patients (groups 1 and 2, 13 patients each, with an ejection fraction > 45% and < 45%, respectively) at 12 selected times. During operation, mean differences (bias) between PCCO-BCO and CCO-BCO did not differ in either group. However, phenylephrine-induced increases in systemic vascular resistance (SVR) by approximately 60% resulted in significant differences. Significantly higher absolute bias values of PCCO-BCO compared with CCO-BCO were also found early after operation in the ICU. Thus PCCO and CCO provided comparable measurements during coronary bypass surgery. After marked changes in SVR, further calibration of the PCCO device is necessary. PMID- 10472218 TI - Comparison of intubating conditions after rapacuronium (Org 9487) and succinylcholine following rapid sequence induction in adult patients. AB - We have assessed intubating conditions provided by rapacuronium (Org 9487) and succinylcholine after rapid sequence induction of anaesthesia in adult patients undergoing elective surgery. We studied 335 patients, ASA I and II, in five centres. Two hundred and thirty-four subjects with normal body weight and 101 obese subjects were allocated randomly to one of four treatment groups differing in the neuromuscular blocking drug administered (rapacuronium 1.5 mg kg-1 or succinylcholine 1 mg kg-1) and in the technique used for induction of anaesthesia (fentanyl 2-3 micrograms kg-1 with thiopental 3-6 mg kg-1 or alfentanil 20 micrograms kg-1 with propofol 1.5-2 mg kg-1). Intubation was started at 50 s by an anaesthetist blinded to the drugs used. Intubating conditions were clinically acceptable (excellent or good) in 89.4% of patients after rapacuronium and in 97.4% after succinylcholine (P = 0.004), the estimated difference being 8.1% (95% confidence interval (CI) 2.0-14.1%). Neither anaesthetic technique nor subject group had an influence on intubating conditions. After intubation, the maximum increase in heart rate averaged 23.1 (SD 25.4%) and 9.4 (26.1%) after rapacuronium and succinylcholine, respectively (P < 0.001). Pulmonary side effects (bronchospasm and increased airway pressure) were observed in 10.7% (95% CI 5.8-17%) and 4.1% (95% CI 1.3-8.8%) of patients given rapacuronium and succinylcholine, respectively (P = 0.021). We conclude that after rapid sequence induction of anaesthesia in adults, clinically acceptable intubating conditions were achieved less frequently after rapacuronium 1.5 mg kg-1 than after succinylcholine. PMID- 10472217 TI - High-dose aprotinin with gentamicin-vancomycin antibiotic prophylaxis increases blood concentrations of creatinine and cystatin C in patients undergoing coronary artery bypass grafting. AB - Both aprotinin and gentamicin-vancomycin antibiotic prophylaxis have been used widely in cardiac surgery to prevent bleeding and infections, respectively. As the drugs are excreted almost entirely by glomerular filtration, we investigated their action on renal function when administered either separately or together. To increase consistency, we measured serum concentrations of creatinine and cystatin C, a new marker of glomerular filtration rate, that many recent studies have shown to be more sensitive than serum creatinine. One hundred patients undergoing coronary artery bypass surgery were allocated randomly to one of four groups: group A received antibiotic prophylaxis with cefamandole and no aprotinin; group B received cefamandole and high-dose aprotinin; group C received antibiotic prophylaxis with gentamicin and vancomycin, but no aprotinin; and group D received both high-dose aprotinin and gentamicin-vancomycin antibiotic prophylaxis. Data from 84 patients, for whom data collection was complete, were analysed. In the first week after operation, mean serum concentrations of cystatin C and creatinine either remained constant or decreased slowly in all groups, except for group D. In group D, both markers increased gradually from postoperative day 2 onwards. The increase in cystatin C was significant on postoperative day 5 (from mean 1.02 (SD 0.11) mg litre-1 before operation to 1.35 (0.32) mg litre-1; P < 0.05), reaching a peak on postoperative day 7 (1.45 (0.35) mg litre-1; P < 0.05), while the increase in creatinine concentration was significant on postoperative day 6 (from 1.05 (0.16) mg dl-1 before operation to 1.29 (0.34) mg dl-1; P < 0.05). We conclude that simultaneous administration of high-dose aprotinin and prophylactic use of gentamicin with vancomycin increased serum concentrations of cystatin C and creatinine in the first postoperative week in patients undergoing cardiac surgery. PMID- 10472219 TI - Influence of metoclopramide on plasma cholinesterase and duration of action of mivacurium. AB - Mivacurium is metabolized by plasma cholinesterase (PCHE). Metoclopramide inhibits PCHE in vitro and in vivo. We have assessed the effect of metoclopramide on duration of action of mivacurium and measured PCHE at baseline and at the time of maximal block. In a randomized, double-blind study, 30 patients received metoclopramide 0.15 mg kg-1 i.v. or saline, followed by propofol anaesthesia and mivacurium 0.15 mg kg-1. Using a TOF-Guard accelerometer, times to recovery of TI to 25%, 75% and 90% were 13.4, 19.3 and 21.9 min in the saline group and 17.8, 25.3 and 28.8 min in the metoclopramide group (P < 0.01, P < 0.05, P < 0.05, respectively). There were no differences in onset time or recovery index between the groups. PCHE activity at the time of maximum block decreased within each group (P < 0.01) but there was no difference between groups. In a second biochemical study of eight patients, a small decrease in PCHE activity was detected after metoclopramide 0.15 mg kg-1, but before administration of mivacurium (P < 0.025). We conclude that metoclopramide prolongs the duration of action of mivacurium. PMID- 10472220 TI - Cardioventilatory coupling: effects of IPPV. AB - Cardioventilatory coupling (CVC) is the temporal coherence of respiratory and cardiac rhythms. We have suggested that this coherence is the result of triggering of inspiratory onset by a preceding cardiovascular afferent. One implication of this triggering hypothesis is that coupling should only exist under conditions of spontaneous ventilation and not under conditions where the ventilatory period is fixed, as during intermittent positive pressure ventilation (IPPV). This study compared the degree of CVC in 20 ASA I female subjects, aged 21-50 yr, 10 of whom were breathing spontaneously and 10 were undergoing mechanical ventilation. Over 5-10 min, we recorded the timing of consecutive ECG R waves and inspiratory onsets. Coupling was demonstrated by examining these epochs for constant timing relationship between R waves and inspiration (RI intervals). Constancy of RI intervals was examined graphically (RI plot) and quantitatively using the Shannon entropy of RI interval distribution. We observed CVC in all spontaneously breathing subjects but in none of those receiving IPPV. In spontaneously breathing subjects, temporal alignment with inspiratory onset was greatest for the heart beat preceding inspiration. We conclude that although coupling has been shown to persist in the presence of electrical cardiac pacing, IPPV disrupts the coupling process, consistent with the view that in anaesthetized subjects, coupling is the triggering of inspiratory onset by a preceding heart beat and not the converse. PMID- 10472221 TI - Dynamics of re-expansion of atelectasis during general anaesthesia. AB - A major cause of impaired gas exchange during general anaesthesia is atelectasis, causing pulmonary shunt. A 'vital capacity' (VC) manoeuvre (i.e. inflation of the lungs up to 40 cm H2O, maintained for 15 s) may re-expand atelectasis and improve oxygenation. However, such a manoeuvre may cause adverse cardiovascular effects. Reducing the time of maximal inflation may improve the margin of safety. The aim of this study was to analyse the change over time in the amount of atelectasis during a VC manoeuvre in 12 anaesthetized adults with healthy lungs. I.v. anaesthesia with controlled mechanical ventilation (VT 9 (SD 1) ml kg-1) was used. For the VC manoeuvre, the lungs were inflated up to an airway pressure (Paw) of 40 cm H2O. This pressure was maintained for 26 s. Atelectasis was assessed by analysis of computed x-ray tomography. The amount of atelectasis, measured at the base of the lungs, was 4.0 (SD 2.7) cm2 after induction of anaesthesia. The decrease in the amount of atelectasis over time during the VC manoeuvre was described by a negative exponential function with a time constant of 2.6 s. At an inspired oxygen concentration of 40%, PaO2 increased from 17.2 (4.0) kPa before to 22.2 (6.0) kPa (P = 0.013) after the VC manoeuvre. Thus in anaesthetized adults undergoing mechanical ventilation with healthy lungs, inflation of the lungs to a Paw of 40 cm H2O, maintained for 7-8 s only, may re expand all previously collapsed lung tissue, as detected by lung computed tomography, and improve oxygenation. We conclude that the previously proposed time for a VC manoeuvre may be halved in such subjects. PMID- 10472222 TI - Sevoflurane anaesthesia with an Oxford Miniature Vaporizer in vaporizer inside circle mode. AB - Anaesthesia was induced and maintained successfully with sevoflurane using an Oxford Miniature Vaporizer (OMV) in vaporizer inside circle (VIC) mode. With continuous monitoring using the Drager Narkomed 4 machine agent analyser, the expired sevoflurane concentration was monitored and controlled easily. During induction, there was no cardiovascular depression, apnoea or coughing, but involuntary movements and breath-holding were common. PMID- 10472223 TI - Sevoflurane requirements for tracheal intubation with and without fentanyl. AB - We studied 80 healthy ASA 1 patients (aged 20-52 yr) to determine if fentanyl affects sevoflurane requirements for achieving 50% probability of no movement in response to laryngoscopy and tracheal intubation (MAC-TI). Patients were allocated randomly to one of four fentanyl dose groups (0, 1, 2 and 4 micrograms kg-1). Patients in each group received sevoflurane at a pre-selected end-tidal concentration according to an 'up-down' technique. After steady state sevoflurane concentration had been maintained for at least 10 min, fentanyl was administered i.v. Tracheal intubation was performed 4 min after administration of fentanyl, and patients were assessed as moving or not moving. Heart rate (HR) and mean arterial pressure (MAP) were recorded before induction of anaesthesia, just before administration of fentanyl, just before laryngoscopy for intubation, and after intubation. The MAC-TI of sevoflurane was 3.55% (95% confidence intervals 3.32-3.78%), and this was reduced markedly to 2.07%, 1.45% and 1.37% by addition of fentanyl 1, 2 and 4 micrograms kg-1, with no significant difference in the reduction between 2 and 4 micrograms kg-1, showing a ceiling effect. Fentanyl attenuated haemodynamic responses (HR and MAP) to tracheal intubation in a dose dependent manner, even with decreasing concomitant sevoflurane concentration. Fentanyl 4 micrograms kg-1 suppressed the changes in HR and MAP more effectively than fentanyl 1 or 2 micrograms kg-1 at sevoflurane concentrations close to MAC TI. PMID- 10472224 TI - Effect of the laryngeal mask airway on oesophageal pH: influence of the volume and pressure inside the cuff. AB - We studied gastro-oesophageal reflux (GOR) with a face mask and laryngeal mask airway (LMA), and the effects of inflation pressure and volume of the LMA cuff on oesophageal pH, in 60 patients. Patients were managed with either a face mask (group I) or LMA inflated to obtain a seal in the anaesthesia circuit at 7 cm H2O (group II) or 15 cm H2O (group III). A pH-sensitive probe with two electrodes, 10 cm apart, was placed in the oesophagus during anaesthesia and recordings were made continuously until patients awakened. There was a significant difference in the incidence of GOR between the face mask (group I) and the LMA (groups II-III) (P < 0.05) in the lower oesophagus but there was no difference in the mid oesophagus. No correlation was found between pressure and volume inside the cuff and variations in oesophageal pH. We conclude that LMA use was associated with increased reflux in the low oesophagus but oesophageal pH was not influenced by variations in pressure or volume inside the LMA cuff. PMID- 10472226 TI - Transient neurological symptoms after spinal anaesthesia with hyperbaric 5% lidocaine or general anaesthesia. AB - Transient neurotoxicity of concentrated local anaesthetics has been thought to be the main reason for transient neurological symptoms after spinal anaesthesia. Profound musculoligamental relaxation by high doses of local anaesthetics may contribute to the development of postoperative musculoskeletal pain. In order to evaluate the role of the loss of strength of the supportive structures of the spine in the development of transient neurological symptoms, 60 patients (ASA I II) undergoing minor orthopaedic, varicose vein or inguinal hernia operations were allocated randomly to receive spinal anaesthesia with hyperbaric lidocaine 50 mg ml-1 (85-100 mg) or balanced general anaesthesia with neuromuscular block. Patients were interviewed 24 h later and after 1 week they returned a written questionnaire. Transient neurological symptoms, consisting of pain in the buttocks or pain radiating symmetrically to the lower extremities, occurred in eight patients (27%) receiving spinal anaesthesia and in one patient (3%) receiving general anaesthesia (P < 0.05). We conclude that a transient neurotoxic effect of hyperbaric lidocaine 50 mg ml-1 is probably the main reason for transient neurological symptoms after spinal anaesthesia but musculoligamental relaxation may contribute to the development of low back or leg pain after both anaesthetic techniques. PMID- 10472225 TI - Oral mucosal blood flow, plasma epinephrine and haemodynamic responses after injection of lidocaine with epinephrine during midazolam sedation and isoflurane anaesthesia. AB - We have investigated the relationship between oral mucosal blood flow and plasma epinephrine concentration, and the effects of conscious sedation vs general anaesthesia on haemodynamic responses after submucosal epinephrine injection in 14 subjects. The same seven patients were studied both as controls and after sedation. For sedation, midazolam i.v. was used. Another seven patients underwent orthognathic surgery with isoflurane anaesthesia. All subjects received a submucosal injection of epinephrine 0.8 microgram kg-1, given as 2% lidocaine hydrochloride with epinephrine 12.5 micrograms ml-1. Baseline mucosal blood flow and peak increase in plasma epinephrine concentration in the general anaesthesia and sedation groups were approximately 2.0 and 1.5 times, respectively, higher than those in the control group. Mean plasma epinephrine concentration reached a maximum 3 min after administration of epinephrine in all groups. Overall, there was a significant correlation (r = 0.65) between baseline mucosal blood flow and the maximum increase in plasma epinephrine concentration. There were no differences in haemodynamic changes except for heart rate, between the three groups. These results suggest that plasma epinephrine concentration after submucosal injection depends on the initial mucosal blood flow in the injected area. Haemodynamic changes, except heart rate, in the sedation and general anaesthesia groups were similar despite different changes in maximum plasma epinephrine concentration. PMID- 10472227 TI - Target-controlled infusion of alfentanil for postoperative analgesia: contribution of plasma protein binding to intra-patient and inter-patient variability. AB - We have examined the influence of plasma protein binding on inter-individual and intra-individual variability in the effective postoperative analgesic concentration (EAC) of alfentanil and on the performance of the target-controlled infusion system used. Ten patients received standardized anaesthesia and target controlled alfentanil for postoperative analgesia. Analgesia was assessed using a visual analogue scale (VAS). Plasma protein binding of alfentanil was assessed at four different times (on arrival in the recovery room, at 21:00 on the day of surgery and at 09:00 and 21:00 on the first postoperative day). Bias and inaccuracy were examined on the day of surgery and on the first postoperative day. Unbound fractions of alfentanil varied from 5 to 15% and varied in time. In general, the unbound fractions on the day of surgery were higher than those on the first postoperative day. Thirty-nine percent of inter-individual variability in the EAC of alfentanil (range 33-140 ng ml-1) at the onset of therapy could be explained by protein binding. At the other observation times, correlations between unbound fraction and EAC were only moderate. Bias on the day of surgery was -19% and 12% on the first postoperative day (ns). Inaccuracy was 23% and 18%, respectively (ns). We conclude that inter-individual variations in plasma protein binding can explain a significant portion of inter-individual variability in the EAC of alfentanil in the early postoperative phase. PMID- 10472228 TI - Combined field block and i.p. instillation of ropivacaine for pain management after laparoscopic sterilization. AB - We have studied the effect of ropivacaine for combined port site and mesosalpinx infiltration, and peritoneal instillation on pain, nausea and vomiting after laparoscopic sterilization, in a double-blind, placebo-controlled study in 80 patients. The total dose of ropivacaine was 285 mg. All patients received intra- and postoperative NSAID in fixed doses. Abdominal and shoulder pain, nausea and vomiting were recorded during the first 8 h after operation and in a diary for 3 days. In the ropivacaine group, abdominal pain scores were lower during the first 4 h (P < 0.00001), additional use of morphine was less (P < 0.001) and fewer patients had nausea or vomiting during the first 72 h (five vs 14; P < 0.05). There were no signs of local anaesthetic toxicity. PMID- 10472229 TI - Is anaesthesia evidence-based? A survey of anaesthetic practice. AB - We were interested in measuring the proportion of anaesthetic interventions in routine practice that are supported by evidence in the literature. We surveyed our hospital practice, asking anaesthetists to nominate a primary problem (if any) and their chosen intervention. Each intervention was classified into one of four levels according to the strength of the evidence recovered from the literature. We found that 96.7% were evidence-based (levels I-IV), including 32% supported by randomized, controlled trials (levels I and II). These results are similar to recent studies in other specialties and refute the claim that only 10 20% of treatments have any scientific foundation. PMID- 10472230 TI - Effects of diazepam and ketamine administered individually or in combination on regional rates of glucose utilization in rat brain. AB - The effects of diazepam, which acts at GABAA receptors to enhance the effects of GABA, and ketamine, a non-competitive N-methyl-D-aspartate receptor antagonist, on local rates of cerebral glucose utilization (ICMRglc) were examined in unrestrained rats. Four groups were studied: vehicle-injected controls; and ketamine-treated, diazepam-treated and combined ketamine- and diazepam-treated animals. Ketamine alone produced a heterogeneous pattern of changes in ICMRglc (e.g. significant increases in the corpus callosum, olfactory tubercle and the entire Papez circuit, in addition to other limbic areas, and significant decreases in lateral habenula and some components of the auditory system). Diazepam alone statistically significantly decreased ICMRglc in the brain as a whole and in most areas of the cerebral cortex, thalamus and limbic system. The most remarkable effects of the two drugs administered together on ICMRglc occurred in the limbic system where the dramatic increases observed with ketamine alone were prevented by treatment with diazepam. PMID- 10472231 TI - Stereospecific effects of ketamine on dopamine efflux and uptake in the rat nucleus accumbens. AB - In addition to being a general anaesthetic, ketamine is a recognized drug of abuse. Many, if not all, drugs of abuse have been shown to increase dopamine efflux in the nucleus accumbens (NAc). As ketamine is optically active, we examined if its actions on dopamine efflux in the NAc were stereoselective. Slices of rat NAc were superfused with artificial CSF at 32 degrees C. Dopamine efflux was evoked by electrical stimulation (1 or 20 pulses, 100 Hz) and measured using fast cyclic voltammetry. (+/-)-Ketamine 100 mumol litre-1 increased dopamine efflux (to mean 174 (SEM 17)% of control, P < 0.05) and slowed dopamine uptake half-time (T1/2) to 164 (17)% of control, as did (+)-ketamine 100 mumol litre-1 (efflux 236 (16)% (P < 0.001); uptake T1/2 177 (25)% (P < 0.05)). The (-) isomer was inactive. The effect of (+)-ketamine on dopamine efflux did not correlate with its action on dopamine uptake. (+)-Ketamine increased dopamine efflux on single pulse stimulation but to a lesser extent than on 20 pulse trains (P < 0.05). (+)-Ketamine was unable to block the inhibitory effect of quinpirole on single pulse dopamine efflux. Neither MK 801 10 mumol litre-1 nor metoclopramide 1 mumol litre-1 had any effect on dopamine release after short train stimuli (20 pulses, 100 Hz). We conclude that the (+)-isomer is the active form of ketamine and increases NAc dopamine efflux not by block of dopamine uptake; autoreceptors or NMDA receptors, but by mobilization of the dopamine storage pool to releasable sites. PMID- 10472232 TI - Mechanisms underlying the inotropic action of halothane on intact rat ventricular myocytes. AB - The mechanisms contributing to the negative inotropic effect of halothane were studied in isolated rate ventricular myocytes. Contraction and intracellular Ca2+ transients were measured optically in these cells. The initial application of halothane (2% or 0.5 mmol litre-1) led to short-lived increases in the Ca2+ transient and contraction, which were abolished by ryanodine. Continued application of halothane led to a sustained decrease in contraction: this resulted from: (i) a decrease in myofilament Ca2+ sensitivity; (ii) a decrease in the Ca2+ transient; and (iii) a decrease in the Ca2+ content of the sarcoplasmic reticulum. Although halothane reduced action potential duration, the sustained negative inotropic effect was similar when action potentials or voltage clamp pulses of constant duration were used to trigger contractions. In cells exposed to nifedipine 0.5 mumol litre-1 (which decreases the L-type Ca2+ current, ICa), Ca2+ transients, sarcoplasmic reticulum Ca2+ content and fractional release (the fraction of sarcoplasmic reticulum Ca2+ content released during each stimulus) were reduced. Halothane 0.5 mmol litre-1 (which also decreases ICa) decreased Ca2+ transients to a lesser extent and reduced sarcoplasmic reticulum Ca2+ content to a greater extent than nifedipine, whereas fractional release was unchanged compared with control. These data suggest that halothane sensitizes Ca(2+)-induced Ca2+ release from the sarcoplasmic reticulum in addition to reducing ICa. PMID- 10472233 TI - Protective effects of anaesthetics in reversible and irreversible ischaemia reperfusion injury. PMID- 10472234 TI - Effect of a bolus dose of midazolam on the auditory evoked response in humans. AB - We have studied the effect of a bolus dose of midazolam on the auditory evoked response (AER) of the electroencephalogram (EEG) in nine patients. We measured the AER in the awake patient, at the point of loss of the eyelash reflex and when airway support was required. The eyelash reflex was lost at mean 1.78 (SD 0.5) min after administration of the midazolam bolus dose. Time to airway support in the seven patients who required it was 2.74 (1.26) min. Mean Nb latency in awake patients was 44.3 ms (95% CI 41.9-46.9) which was significantly shorter than Nb latency at the clinical end-points (P < 0.001). When the eyelash reflex was lost, Nb latency was 55.7 ms (95% CI 51.4-60.3) and when airway support was needed, it was 50.9 ms (95% CI 48.6-53.2). We conclude that loss of consciousness after midazolam was associated with an increase in mean Nb latency. PMID- 10472235 TI - Patient-controlled sedation using propofol in elderly patients in day-case cataract surgery. AB - Patient-controlled sedation (PCS) with propofol has been used successfully as an adjunct to local anaesthetic procedures. We studied a group of elderly patients (mean age 75.4 yr) undergoing cataract surgery and attempted to increase patient acceptability and comfort of local anaesthesia. Propofol was self-administered in a dose of 0.25 mg kg-1 for patients more than 60 yr of age, with a lockout period of 3 min. A total of 14 of 20 patients used PCS; eight of 20 used the PCS only once and another six had three tries or less. Despite this, 18 of 20 patients claimed they found the PCS useful. However, while it is possible to administer PCS successfully to elderly patients undergoing cataract surgery and produce a decrease in the level of anxiety, we found it unacceptable because of head movement in two patients. These patients received only two and three divided doses, to a maximum of 29 and 30 mg, respectively. There were no other adverse events. PMID- 10472236 TI - Tetracaine gel vs EMLA cream for percutaneous anaesthesia in children. AB - We have evaluated the anaesthetic effect of tetracaine gel 1 g, applied for 45 min, compared with EMLA cream 2 g, applied for 60 min, in a randomized, double blind study in 60 children aged 3-15 yr. Venous cannulation was performed 15 min after removal of the EMLA cream (n = 20) and tetracaine gel (n = 20). Cannulation was performed up to 215 min after removal of the tetracaine gel in another 20 patients. Significantly lower pain scores were recorded by the children treated with tetracaine gel compared with EMLA cream (P < 0.02). Forty to 45% of children in the tetracaine groups reported no pain compared with only 10% in the EMLA group. Only minor adverse effects were observed. We conclude that tetracaine gel provided effective, rapid, long-lasting and safe local anaesthesia, and was significantly better than EMLA cream in reducing pain during venous cannulation in children using the recommended application periods for both formulations. PMID- 10472237 TI - I.v. clonidine: does it work as a hypotensive agent with inhalation anaesthesia? AB - In a double-blind, randomized, placebo-controlled study, 41 patients received clonidine 3 micrograms kg-1 or placebo at induction of isoflurane and nitrous oxide in oxygen anaesthesia. Metoprolol was also given to achieve a systolic arterial pressure of 80 mm Hg. Requirements for metoprolol were significantly less in the clonidine group (P < 0.00035), with no significant difference in mean arterial pressures over time. It would appear that clonidine is an i.v. hypotensive agent worthy of consideration, but data during the recovery period are required to comment further on the safety of this technique. PMID- 10472238 TI - Measurement of peripheral tissue thickness by ultrasound during the perioperative period. AB - We have studied changes in peripheral tissue thickness with a novel hand-held ultrasound device during the perioperative course of 60 healthy surgical patients in three different intraoperative body positions. The nil-by-mouth period led to a significant decrease in forehead tissue thickness. Standardized infusion therapy with Ringer's solution at a rate of 8 ml kg-1 h-1 resulted in a gradual increase in tissue thickness, which was significantly different from preoperative baseline values after 90 min. Packed cell volume decreased significantly after the start of infusion and remained low over the rest of the observation time. Different body positions did not influence changes in tissue thickness. We conclude that changes in perioperative tissue thickness in healthy patients can be detected easily by ultrasound, independent of body position. This method may prove useful for the non-invasive assessment of fluid balance state. PMID- 10472239 TI - Severe postoperative laryngeal oedema causing total airway obstruction immediately on extubation. AB - We report a case of total upper airway obstruction occurring immediately after extubation after elective bi-maxillary osteotomy. The obstruction was caused by severe, progressive supraglottic oedema, which totally obscured the laryngeal inlet. No swelling had been present at initial laryngoscopy and intubation. Immediate re-intubation of the patient's trachea was difficult but life saving. Subsequent investigations revealed extensive soft tissue swelling, maximal at the level of the hyoid and extending downwards into the trachea. The cause of such severe oedema in this case is not certain, but may be related to vigorous submental liposuction carried out at the end of operation. We have found no other reports of total airway obstruction occurring immediately after extubation as a result of this cause. We review the appropriate literature, describe the postoperative management and suggest precautions in similar patients. PMID- 10472240 TI - Proportional and continuous NO delivery systems. PMID- 10472241 TI - Consciousness and molecular mechanisms of anaesthesia. PMID- 10472243 TI - Mental nerve neuropraxia associated with tracheal intubation using an RAE tube. PMID- 10472242 TI - Intubating conditions after propofol and remifentanil. PMID- 10472244 TI - Hazards of laparoscopic surgery. PMID- 10472245 TI - Ropivacaine and bupivacaine for analgesia in labour. PMID- 10472246 TI - The practical application and interpretation of simple lung function tests in cystic fibrosis. PMID- 10472247 TI - Advantages of comprehensive lung function techniques in cystic fibrosis. PMID- 10472248 TI - Oxygen therapy for cystic fibrosis. PMID- 10472250 TI - Assessment for lung transplantation in cystic fibrosis. PMID- 10472249 TI - The protease-antiprotease battle in the cystic fibrosis lung. PMID- 10472251 TI - Diabetes in cystic fibrosis. PMID- 10472253 TI - The role of the general practitioner in cystic fibrosis. PMID- 10472252 TI - Update on intestinal strictures. PMID- 10472254 TI - Attention deficit hyperactivity disorder. PMID- 10472255 TI - Roots of violence. PMID- 10472256 TI - Who is suitable for cognitive behavioural therapy? PMID- 10472257 TI - Unproven assumptions about the impact of bereavement on children. PMID- 10472258 TI - Iron deficiency in childhood. PMID- 10472259 TI - No association of Chlamydia with abortion. AB - Case reports and serological work have raised the possibility that chlamydias can infect the placenta and thus harm the fetus. We investigated the involvement of Chlamydia in a series of 195 unselected cases of spontaneous abortion or miscarriage. Formalin-fixed placental tissues from all cases were examined immunohistochemically, for the presence of chlamydial lipopolysaccharide, as well as histopathologically. A serum sample was collected from 187 of the patients for detection of anti-chlamydial antibodies by microimmunofluorescence. All placental sections were negative for chlamydial antigen. Serological findings indicated that 8 patients had been in contact with C. trachomatis, 15 patients with C. pneumoniae, and none with C. psittaci. A few cases of perivillitis or intervillitis were detected, but none exhibited the intracytoplasmic inclusions typical of C. psittaci. Although these results are negative a search for Chlamydia in abortion materials should be encouraged. PMID- 10472260 TI - Two-surgeon approach to thoracic outlet syndrome: long-term outcome. AB - An orthopaedic surgeon and a vascular surgeon jointly conducted 30 operations for thoracic outlet syndrome in 27 patients, having done the preoperative assessments in conjunction with a neurologist. Anterior scalenectomy was performed by the supraclavicular route except in one case where the infraclavicular route was used. The further surgical procedure was tailored to the abnormalities identified -i.e. resection of cervical rib or band, or medial scalenectomy. The first rib was spared. At median follow-up of 37 months (range 3-228) results were judged excellent or good on 26/30 sides (87%); on the three occasions when scalenectomy alone was performed, the results were only fair or poor. There were no major complications and no patient required reoperation. The long-term outcome in this series suggests that, with multidisciplinary assessment and two-surgeon operative treatment, good results can be obtained by the supraclavicular route without resection of the first rib. PMID- 10472261 TI - Orthopaedic management in four cases of mucolipidosis type III. AB - Four patients with mucolipidosis type III, three of them brothers, were seen initially in the first two decades of life. Their main symptoms were carpal tunnel syndrome, trigger fingers and generalized joint stiffness. Radiographs showed spinal deformities and hip dysplasia, but these were not causing pain. Carpal tunnel syndrome was treated surgically but joint stiffness and hip and knee contractures were managed by physiotherapy. Up to the age of 24 none of these patients has had pelvic osteotomy for hip dysplasia; this operation, not yet reported in mucolipidosis type III, may eventually be necessary. PMID- 10472262 TI - Non-communication between ophthalmologists and optometrists. AB - Many patients seen in the British hospital eye service are referred by high street optometrists; and, if the optometrist is to receive feedback from the ophthalmologist, the patient should consent to disclosure of medical information. On the referral form (revised GOS 18) there is a space for this purpose. We investigated the level of communication by asking optometrists in our hospital catchment area about their use of the GOS 18 form and by examining the medical records of all new patients seen in the eye outpatient department in one month. 79 optometrists (55%) returned the questionnaire. 54 routinely used the GOS 18; and, of these, 10 said they obtained patient consent always, 23 sometimes and 21 never. 158 of 555 sets of medical notes contained an optometrist's referral, 107 of them on the revised GOS 18; and patient consent had been recorded on 17 of these forms. Ophthalmologists responded to the optometrist in 2/17 (12%) cases where consent had been obtained and 15/90 (17%) where it had not. Ophthalmologists could provide much better feedback to optometrists. The GOS 18 form could be used more effectively; and there is no reason why patient consent to disclosure of medical information should not be obtained by ophthalmologists as well as by optometrists. PMID- 10472263 TI - A tooth for an eye: dental procedures in unrecognized glaucoma. PMID- 10472264 TI - Dental prosthesis in the retropharyngeal space. PMID- 10472265 TI - Ultrasound appearances in chronic appendicitis. PMID- 10472266 TI - Double pylorus as a cause of gastrointestinal bleeding. PMID- 10472267 TI - Duodenal diverticulum as a cause of massive gastrointestinal bleeding. PMID- 10472268 TI - Mycotic aneurysm after percutaneous transluminal femoral artery angioplasty. PMID- 10472269 TI - Peritonsillar abscess after self-cleaning of tonsillar remnant. PMID- 10472270 TI - Tobias Smollett consults a French physician in 1763. PMID- 10472271 TI - Mixed signals. PMID- 10472272 TI - Palliative medicine as a specialty. PMID- 10472274 TI - Extinction of the general physician. PMID- 10472273 TI - Recreational pharmacology. PMID- 10472275 TI - Music as male competition. PMID- 10472277 TI - Community institutional healthcare: emergence from refugee status. PMID- 10472278 TI - The dilemma of antenatal HIV testing: what goes on in the European Community? PMID- 10472279 TI - Trends in teenage pregnancy in England and Wales: how can we explain them? AB - Teenage pregnancy is associated with adverse social and physical outcomes for both mother and child. We drew on various sources--birth and abortion statistics from the Office for National Statistics, data from the National Survey of Sexual Attitudes and Lifestyles, and routinely collected data from family planning clinics--to identify trends in England and Wales and their possible determinants. The rate of teenage sexual activity has increased steadily and consistently over the past four decades, whilst the rate of teenage fertility has shown greater variation. When the teenage fertility rate is calculated against the denominator of sexually active women, rather than the total sample of teenage women, the underlying trend in teenage fertility over the past four decades has been downwards, though not consistently so. Fluctuations in the teenage fertility rate seem to track intervention-related factors such as access to, and use of, contraceptive services and the general climate surrounding the sexual health of young people. PMID- 10472280 TI - Antibacterial activity of honey against strains of Staphylococcus aureus from infected wounds. AB - The antibacterial action of honey in infected wounds does not depend wholly on its high osmolarity. We tested the sensitivity of 58 strains of coagulase positive Staphylococcus aureus, isolated from infected wounds, to a pasture honey and a manuka honey. There was little variation between the isolates in their sensitivity to honey: minimum inhibitory concentrations were all between 2 and 3% (v/v) for the manuka honey and between 3 and 4% for the pasture honey. Thus, these honeys would prevent growth of S. aureus if diluted by body fluids a further seven-fold to fourteen-fold beyond the point where their osmolarity ceased to be completely inhibitory. The antibacterial action of the pasture honey relied on release of hydrogen peroxide, which in vivo might be reduced by catalase activity in tissues or blood. The action of manuka honey stems partly from a phytochemical component, so this type of honey might be more effective in vivo. Comparative clinical trials with standardized honeys are needed. PMID- 10472281 TI - Coeliac disease in adults: variations on a theme. AB - In childhood, coeliac disease (gluten enteropathy) tends to show itself with failure to thrive and growth retardation; in adult life with malabsorption syndromes. We report six cases in adults who presented atypically, with features including clotting disorder, hypoglycaemia, weight loss, anaemia and angina pectoris, all of which responded to gluten withdrawal. PMID- 10472282 TI - Consent to treatment and the mentally incapacitated adult. AB - Doctors are sometimes faced with adult patients who lack the mental capacity to consent to treatment. In a questionnaire, 120 doctors in a district general hospital were asked what action they would take if such a patient had a clear need for elective treatment. Of the 89 who replied, 57 said they would seek consent from relatives or others; 11 of these, nevertheless, stated that treatment could proceed without such consent. These results, and inquiries about other options, pointed to widespread misunderstanding of the law. In English law, no one can give legally valid consent on behalf of another adult. When an individual is unable to give consent, common law allows a doctor to protect a patient's best interests by treating him or her in accordance with a responsible body of medical opinion. PMID- 10472283 TI - Clinical experience during the paediatric undergraduate course. AB - Medical students at the Cambridge Clinical School are provided with a list of 42 core conditions they should encounter and 20 core skills they should perform during their attachment. By self-completion questionnaires we assessed their clinical experience and the amount of teaching they received, relating the results to marks gained in end-of-attachment assessments. 103 (93%) of 110 students in year one and 123 (96%) of 128 in year two completed the questionnaires. Of the 42 core conditions, 13 were seen by under 70% of the students in year one. In year two, exposure rate increased for 26 core conditions by a median of 7% (range 2-40) and decreased in 13 core conditions by a median value 4% (range 5-13) (P = 0.0005, chi 2). Only mandatory core skills were performed by over 90% of students. 5% of students did not perform any newborn examinations and under 60% observed neonatal resuscitation or a high-risk delivery. Students' core condition score was associated with their core skill score (r = 0.5), hospital grade (r = 0.3) and exposure to acute paediatrics (r = 0.3) (P < 0.005). There was no significant association between clinical experience and the objective examination score or the amount of teaching received. There was an inverse association between the number of students at a hospital and the number of core conditions with an exposure rate above 70% at that hospital (r = 0.7, P < 0.05). This study suggests that clinical experience may be better judged by the clinical supervisor than by assessment of theoretical knowledge. PMID- 10472284 TI - Restoration of voice after laryngectomy. PMID- 10472285 TI - Neurological symptoms 27 years after tiger bite. PMID- 10472286 TI - Neck pain with fever. PMID- 10472287 TI - Streptococcus viridans vertebral osteomyelitis. PMID- 10472288 TI - Gastrointestinal haemorrhage after abdominal aortic grafting. PMID- 10472289 TI - Abdominal mass in a woman taking non-steroidal anti-inflammatory drugs. PMID- 10472290 TI - Recurrent transitional cell carcinoma in a Boari flap. PMID- 10472291 TI - Autoimmune enteropathy with goblet-cell antibodies. PMID- 10472292 TI - Robert Willan MD FRS (1757-1812): dermatologist of the millennium. PMID- 10472293 TI - Career challenges for consultants. PMID- 10472294 TI - Endocarditis after blood transfusion? PMID- 10472295 TI - Tropical medicine. PMID- 10472296 TI - Hospital consultants' views on homoeopathy. PMID- 10472297 TI - Diaphragmatic rupture. PMID- 10472298 TI - Colonoscopy: if a patient says stop, should we continue? PMID- 10472299 TI - General Dental Council's guidance on anaesthesia. PMID- 10472300 TI - Adverse reactions. PMID- 10472301 TI - Vehicle accidents related to sleep: a review. AB - Falling asleep while driving accounts for a considerable proportion of vehicle accidents under monotonous driving conditions. Many of these accidents are related to work--for example, drivers of lorries, goods vehicles, and company cars. Time of day (circadian) effects are profound, with sleepiness being particularly evident during night shift work, and driving home afterwards. Circadian factors are as important in determining driver sleepiness as is the duration of the drive, but only duration of the drive is built into legislation protecting professional drivers. Older drivers are also vulnerable to sleepiness in the mid-afternoon. Possible pathological causes of driver sleepiness are discussed, but there is little evidence that this factor contributes greatly to the accident statistics. Sleep does not occur spontaneously without warning. Drivers falling asleep are unlikely to recollect having done so, but will be aware of the precursory state of increasing sleepiness; probably reaching a state of fighting off sleep before an accident. Self awareness of sleepiness is a better method for alerting the driver than automatic sleepiness detectors in the vehicle. None of these have been proved to be reliable and most have shortcomings. Putative counter measures to sleepiness, adopted during continued driving (cold air, use of car radio) are only effective for a short time. The only safe counter measure to driver sleepiness, particularly when the driver reaches the stage of fighting sleep, is to stop driving, and--for example, take a 30 minute break encompassing a short (< 15 minute) nap or coffee (about 150 mg caffeine), which are very effective particularly if taken together. Exercise is of little use. CONCLUSIONS: More education of employers and employees is needed about planning journeys, the dangers of driving while sleepy, and driving at vulnerable times of the day. PMID- 10472302 TI - Acquisition and extinction of somatic symptoms in response to odours: a Pavlovian paradigm relevant to multiple chemical sensitivity. AB - OBJECTIVES: Multiple chemical sensitivity is a poorly understood syndrome in which various symptoms are triggered by chemically unrelated, but often odorous substances, at doses below those known to be harmful. This study focuses on the process of pavlovian acquisition and extinction of somatic symptoms triggered by odours. METHODS: Diluted ammonia and butyric acid were odorous conditioned stimuli (CS). The unconditioned stimulus (US) was 7.4% CO2 enriched air. One odour (CS+) was presented together with the US for 2 minutes (CS+ trial), and the other odour (CS-) was presented with air (CS-trial). Three CS+ and three CS exposures were run in a semi-randomised order; this as the acquisition (conditioning) phase. To test the effect of the conditioning, each subject then had one CS+ only--that is, CS+ without CO2--and one CS- test exposure. Next, half the subjects (n = 32) received five additional CS+ only exposures (extinction group), while the other half received five exposures to breathing air (wait group). Finally, all subjects got one CS+ only test exposure to test the effect of the extinction. Ventilatory responses were measured during and somatic symptoms after each exposure. RESULTS: More symptoms were reported upon exposure to CS+ only than to CS-odours, regardless of the odour type. Altered respiratory rate was only found when ammonia was CS+. Five extinction trials were sufficient to reduce the level of acquired symptoms. CONCLUSION: Subjects can acquire somatic symptoms and altered respiratory behaviour in response to harmless, but odorous chemical substances, if these odours have been associated with a physiological challenge that originally had caused these symptoms. The conditioned symptoms can subsequently be reduced in an extinction procedure. The study further supports the plausibility of a pavlovian conditioning hypothesis to explain the pathogenesis of MCS. PMID- 10472303 TI - Work characteristics predict psychiatric disorder: prospective results from the Whitehall II Study. AB - OBJECTIVES: The impact of work on the risk of future psychiatric disorder has been examined in few longitudinal studies. This was examined prospectively in a large epidemiological study of civil servants. METHODS: In the Whitehall II study, a longitudinal, prospective cohort study of 6895 male and 3413 female London based civil servants, work characteristics measured at baseline (phase 1: 1985-8) and first follow up (phase 2: 1989) were used to predict psychiatric disorder measured by a 30 item general health questionnaire (GHQ) at phase 2 and phase 3 follow up (phase 3: 1991-3). Work characteristics and GHQ were measured at all three phases. RESULTS: Low social support at work and low decision authority, high job demands and effort-reward imbalance were associated with increased risk of psychiatric disorder as assessed by the GHQ at follow up adjusting for age, employment grade, and baseline GHQ score. CONCLUSIONS: Social support and control at work protect mental health while high job demands and effort-reward imbalance are risk factors for future psychiatric disorder. Intervention at the level of work design, organisation, and management might have positive effects on mental health in working populations. PMID- 10472304 TI - Incidence of childhood brain and other non-haematopoietic neoplasms near nuclear sites in Scotland, 1975-94. AB - OBJECTIVES: To examine the risk of cancers other than leukaemia and non-Hodgkin's lymphoma in children resident in the vicinity of nuclear sites in Scotland. METHODS: The study dataset comprised registrations of cancer other than leukaemia and non-Hodgkin's lymphoma diagnosed in children aged under 15 in the period 1975 94. These were validated for completeness and accuracy and analysed in two groups: (a) tumours of the central nervous system and (b) other malignant tumours (excluding leukaemia and non-Hodgkin's lymphoma). Around each nuclear site observed cases (O) were enumerated and expected numbers (E) calculated with adjustment for age, sex, deprivation, and an urban-rural category. Stone's maximum likelihood ratio test (MLR) was used to determine whether there was any evidence of increased risk of these neoplasms among children living within 25 km of one of the nuclear sites investigated. The significance level of each MLR statistic was estimated by simulation. RESULTS: More tumours of the central nervous system were observed than expected within 25 km of Dounreay (O/E = 1.14), Hunterston (1.14), and Rosyth (1.22). These results were based on 2, 26, and 136 observed cases, respectively. The unconditional MLR was significant only for Rosyth (p = 0.006). The conditional application of the MLR test for Rosyth was not significant (p = 0.771). For the group of other malignant neoplasms, the unconditional MLR test was not significant for any of the seven sites. CONCLUSIONS: There was no evidence for generally increased risk of either tumours of the central nervous system or other malignant tumours in children living near nuclear sites. The significant excess of tumours of the central nervous system around Rosyth is likely to be due to the high incidence of these tumours in east central Scotland. Further investigations in this area are warranted. PMID- 10472305 TI - Cohort mortality study of 57,000 painters and other union members: a 15 year update. AB - OBJECTIVES: To study mortality patterns in the largest existing cohort of painters. METHODS: 15 years of follow up were added to a study of 42,170 painters and 14,316 non-painters based on union records. There were 23,458 deaths, compared with 5313 in the earlier follow up. RESULTS: Comparisons with the United States population showed significantly increased rates in painters for lung cancer (standardised mortality ratio (SMR) 1.23, 95% confidence interval (95% CI) 1.17 to 1.29), bladder cancer (SMR 1.23, 95% CI 1.05 to 1.43), liver cancer (SMR 1.25, 95% CI 1.03 to 1.50), and stomach cancer (SMR 1.39, 95% CI 1.20 to 1.59). However, in direct comparisons with non-painters only the excesses for lung cancer (SRR 1.23, 95% CI 1.11 to 1.35, increasing to 1.32, 95% CI 16 to 1.93 with 20 years latency) and bladder cancer (SRR 1.77, 95% CI 1.13 to 2.77) were confirmed. Some confounding by smoking may affect these two outcomes, particularly with external referents. Cirrhosis of the liver was increased for both painters and non-painters (SMRs 1.21, 95% CI 1.07 to 1.35, and 1.26, 95% CI 1.03 to 1.51, respectively), possibly indicating high alcohol consumption. Suicide (SMR 1.21, 95% CI 1.05 to 1.38) and homicide (SMR 1.36, 95% CI 1.04 to 1.75) were increased for painters but not for non-painters; neuropsychiatric diseases have been associated with painters in earlier studies. CONCLUSIONS: The results suggest modest occupational risks for lung and bladder cancer; these results are consistent with existing publications. The International Agency for Research on Cancer has classified painting as an occupation definitely associated with cancer. PMID- 10472306 TI - Occupational risk factors of lung cancer: a hospital based case-control study. AB - OBJECTIVES: To investigate the relation between lung cancer and exposure to occupational carcinogens in a highly industrialised region in western Europe. METHODS: In a case-control study 478 cases and 536 controls, recruited from 10 hospitals in the Antwerp region, were interviewed. Cases were male patients with histologically confirmed lung cancer; controls were male patients without cancer or primary lung diseases. Data were collected by questionnaires to obtain information on occupations, exposures, and smoking history. Job titles were coded with the Office of Populations, Censuses and Surveys industrial classification. Exposure was assessed by self report and by job-task exposure matrix. Exposure odds ratios were calculated with logistic regression analysis adjusted for age, smoking history, and marital and socio-economic status. RESULTS: A job history in the categories manufacturing of transport equipment other than automobiles (for example, shipyard workers), transport support services (for example, dockers), and manufacturing of metal goods (for example, welders) was significantly associated with lung cancer (odds ratios (ORs) 2.3, 1.6, and 1.6 respectively). These associations were independent of smoking, education, civil, and economic status. Self reported exposure to potential carcinogens did not show significant associations with lung cancer, probably due to nondifferential misclassification. When assessed by job-task exposure matrix, exposure to molybdenum, mineral oils, and chromium were significantly associated with lung cancer. A strong association existed between smoking and lung cancer: OR of ex-smokers 4.2, OR of current smokers 14.5 v non-smokers. However, smoking did not confound the relation between occupational exposure and lung cancer. CONCLUSIONS: The study has shown a significant excess risk of lung cancer among workers in manufacturing of metal goods, manufacturing of transport equipment (other than automobiles), and transport support services. Assessment of exposure to specific carcinogens resulted in significant associations of chromium, mineral oils, and molybdenum with lung cancer. This study is, to our knowledge, the first study reporting a significant association between occupational exposure to molybdenum and lung cancer. PMID- 10472307 TI - Two year follow up of pulmonary function values among welders in New Zealand. AB - OBJECTIVES: To examine whether welding is a risk factor for an accelerated decline in pulmonary function. METHODS: 2 Year follow up of pulmonary function and respiratory symptoms among 54 welders and 38 non-welders in eight New Zealand welding sites. RESULTS: There were no significant differences in age, height, smoking habits, ethnicity, or total time in industrial work between welders and non-welders. No overall differences were noted in the changes of pulmonary function variables between the two study groups. However, when the comparison was restricted to smokers, welders had a significantly greater (p = 0.02) annual decline (88.8 ml) in FEV1 than non-welders, who had a slight non-significant annual increase (34.2 ml). Also, welders without respiratory protection or local exhaust ventilation while welding had a greater annual decline both in forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) than welders with protection (p = 0.001 and 0.04, respectively). Among welders a significant association was found between the acute across shift change and the annual decline in FEV1. Chronic bronchitis was more common among welders (24%) than non-welders (5%). Only one welder (2%) but eight non-welders (21%) reported having asthma. CONCLUSIONS: Welders who smoked and welders working without local exhaust ventilation or respiratory protection have an increased risk of accelerated decline in FEV1. PMID- 10472308 TI - Chronic rhinitis in workers at risk of reactive airways dysfunction syndrome due to exposure to chlorine. AB - BACKGROUND: To assess the frequency of chronic upper airways symptoms and to relate the presence of these symptoms to accidental exposure to chlorine and changes in lower airways symptoms, airway function, and bronchial responsiveness in a cohort of workers at risk of sporadic occupational exposure to high concentrations of chlorine. METHODS: Data were collected on symptom assessment, spirometry, and methacholine challenge tests from 211 workers seen twice at a 2 year interval (1992-4). RESULTS: The proportion of workers reporting chronic rhinitis was 46.9% in 1992 and 42.2% in 1994. Chronic rhinitis reported in 1994 was significantly associated with acute exposure to chlorine (self reports, p = 0.02; first aid reports, p = 0.001). In a multivariate logistic regression analysis the presence of reported accidents at the first aid unit (one accident, odds ratio (OR) 3.1, 95% confidence interval (95% CI) 1.3 to 7.5; two or more accidents, OR 6.2, 1.1 to 35.8) and of personal atopy (OR 5.5, 2.2 to 10.8) were significant predictors of chronic rhinitis in 1994. Chronic lower airways symptoms were more frequent in 1994 among workers reporting chronic rhinitis on both assessments than in others (p = 0.03) and changes in bronchial responsiveness were more pronounced in those with persistent rhinitis (p = 0.09). CONCLUSIONS: These results suggest that persistent nasal symptoms in workers at risk of reactive airways dysfunction syndrome could be a useful marker of lower respiratory tract abnormalities. PMID- 10472309 TI - Relation between job strain and myocardial infarction: a case-control study. AB - OBJECTIVES: To study the influence of different job related and socioeconomic factors for development of myocardial infarction (MI). METHOD: The study was a case-control study of 76 male wage earners who had been admitted to hospital with MI. As a control group 176 male wage earners not admitted to hospital who were residents of the same county were used. Both groups were interviewed with an extensive questionnaire on job related conditions. Several indices on job related psychosocial factors were established in accordance with Karasek's job strain model as well as the extension of the model, the isostrain model. RESULTS: The most significant findings were consistent with Karasek's job strain model in that mean with a high degree of demand combined with a low degree of control at work had a significantly increased odds ratio (OR) 95% confidence interval (95% CI) of 2.1 (1.2 to 3.8) for MI after adjustment for age compared with men with a low degree of demand and a high degree of control at work. Further adjustment for smoking, socioeconomic status, employment sector, job category, and social network did not affect the OR substantially (OR 2.3 (1.2 to 4.4)). Other factors significantly associated to MI were job category (blue collar workers v white collar workers, OR 2.8 (1.6 to 5.8)), and employment sector (private v public, OR 3.1 (1.8 to 6.1)). CONCLUSIONS: Thus, the study confirmed the job strain model as well as the well known association between socioeconomic status and risk of MI, whereas the finding of an increased risk among employees in the private sector has not previously been described. PMID- 10472310 TI - Interactions between physical and psychosocial risk factors at work increase the risk of back disorders: an epidemiological approach. AB - OBJECTIVES: To investigate the possible interactions between physical and psychosocial risk factors at work that may be associated with self reported back disorders. METHODS: 891 of 1514 manual workers, delivery drivers, technicians, customer services computer operators, and general office staff reported risk factors at work and back disorders with a self administered questionnaire (59% return rate). Of the 869 respondents with a valid questionnaire, 638 workers were classified in to one of four exposure groups: high physical and high psychosocial; high physical and low psychosocial; low physical and high psychosocial; and low physical and low psychosocial. Low physical and low psychosocial was used as an internal reference group. The exposure criteria were derived from existing epidemiological publications and models for physical and psychosocial work factors. The frequency and amplitude of lifting and the duration spent sitting while experiencing vibration were used as physical exposure criteria. Ordinal values of mental demands, job control, and social support from managers and coworkers were used as psychosocial exposure criteria. RESULTS: The highest increase in risk was found in the high physical and high psychosocial exposure group for symptoms of back disorders. In the crude and multivariate analyses, a departure from an additive risk model was found for the 7 day prevalence of symptoms of a low back disorder and also for a recurrent back disorder not present before the current job but also experienced in the past 7 days. CONCLUSION: This study suggests that an interaction between physical and psychosocial risk factors at work may exist to increase the risk of self reported back disorders. Ergonomic prevention strategies that aim to minimise the risks of symptoms of work related back disorders should not only focus on physical but also on psychosocial risk factors at work. The greatest benefits are likely to be realised when both physical and psychosocial factors are put right. PMID- 10472311 TI - Health effects among workers in sewage treatment plants. AB - OBJECTIVES: To further assess the presence of fatigue, symptoms of diarrhoea, and inflammation of airways among people working in sewage plants and the relation to airborne bacterial endotoxin at the workplace. METHODS: 34 Employees in sewage treatment plants and 35 controls were selected. They underwent a questionnaire investigation, and spirometry and airway responsiveness were measured. Measurements were made of airborne endotoxin at different workplaces. RESULTS: The amount of airborne endotoxin varied between 3.8 and 32,170 ng/m3. Workers reported significantly higher nose irritation, tiredness, and diarrhoea. Airway responsiveness was increased among sewage workers, but no differences between the groups were found for spirometry. CONCLUSIONS: The results confirm previous studies on the presence of airways and intestinal inflammation among workers in sewage treatment plants. The most likely causative agent is endotoxin, and at 14 of 23 workplaces, concentrations exceeded recommended guidelines. PMID- 10472312 TI - Genetic damage in operating room personnel exposed to isoflurane and nitrous oxide. AB - OBJECTIVES: To evaluate genetic damage as the frequency of sister chromatid exchanges and micronuclei in lymphocytes of peripheral blood of operating room personnel exposed to waste anaesthetic gases. METHODS: Occupational exposure was measured with a direct reading instrument. Venous blood samples were drawn from 10 non-smokers working in the operating room and 10 non-smoking controls (matched by age, sex, and smoking habits). Lymphocytes were cultured separately over 72 hours for each assay with standard protocols. At the end of the culture time, the cells were harvested, stained, and coded for blind scoring. The exchanges of DNA material were evaluated by counting the number of sister chromatid exchanges in 30 metaphases per probe or by counting the frequency of micronuclei in 2000 binucleated cells. Also, the mitotic and proliferative indices were measured. RESULTS: The operating room personnel at the hospital were exposed to an 8 hour time weighted average of 12.8 ppm nitrous oxide and 5.3 ppm isoflurane. The mean (SD) frequency of sister chromatid exchanges was significantly higher (10.2 (1.9) v 7.4 (2.4)) in exposed workers than controls (p = 0.036) the proportion of micronuclei (micronuclei/500 binucleated cells) was also higher (8.7 (2.9) v 6.8 (2.5)), but was not significant (p = 0.10). CONCLUSION: Exposure even to trace concentrations of waste anaesthetic gases may cause dose-dependent genetic damage. Concerning the micronuclei test, no clastogenic potential could be detected after average chronic exposure to waste anaesthetic gas. However, an increased frequency of sister chromatid exchanges in human lymphocytes could be detected. Although the measured differences were low, they were comparable with smoking 11-20 cigarettes a day. Due to these findings, the increased proportion of micronuclei and rates of sister chromatid exchanges may be relevant long term and need further investigation. PMID- 10472313 TI - Are the children of fathers whose jobs involve contact with many people at an increased risk of leukaemia? AB - OBJECTIVES: To investigate the hypothesis that children of men whose jobs involve contact with many people (particularly children) are at an increased risk of leukaemia. METHODS: A population based dataset obtained from routinely collected death certificates involving 14,168 cancer deaths occurring before the age of 15 years registered in England and Wales between 1959-63 and 1970-90. Associations were assessed with the proportional cancer mortality ratio (PCMR), with all childhood cancer deaths forming the standard for comparison. The PCMRs were adjusted, by stratification, for age and year of death (in 1-year bands) and paternal social class (nine categories). Analyses were performed by estimated level of paternal occupational social contact (high, medium, and low) for all leukaemias, leukaemia subtype, age at death, year of death, and individual occupation. RESULTS: Out of 223 occupations, 36 (16%) were identified as having potentially high levels of social contact, and 27 (12%) as having potentially medium levels of social contact. No associations were found between paternal occupational social contact and death during childhood from leukaemia (high social contact: PCMR 94, 95% confidence interval (95% CI) 87 to 102; medium social contact: 101, 95 to 106). No associations were found when the data were analysed by leukaemia subtype, age at death, year of death, or individual occupation. CONCLUSION: The findings presented here do not support the suggestion that childhood leukaemia is related to the amount of social contact that fathers experience at work. PMID- 10472314 TI - Exposure to toluene in the printing industry is associated with subfecundity in women but not in men. AB - OBJECTIVE: To examine the possible influence of exposure to toluene on human fertility. METHODS: In a cross sectional study, a sample of 150 male and 90 female printing industry workers were interviewed retrospectively on reproductive experience with a modified version of the European study of infertility and subfecundity questionnaire. Exposure categories comprised job descriptions and information on exposure measurements obtained by industrial hygienists. The fecundability ratio (FR) was estimated on the basis of time to pregnancy (TTP) or periods of unprotected intercourse not leading to pregnancy (PUNP) by means of survival analysis with proportional hazard models. Confounders such as age, ethnicity, smoking, parity, pelvic inflammatory diseases, and frequency of sexual intercourse were controlled for in the analyses. RESULTS: 256 Periods of TTP or PUNP were reported by men and 174 by women. After exclusion of induced abortions, birth control failures, and periods without employment for female workers we were able to analyse 169 periods in men and 100 periods in women. Male workers who had been exposed to different concentrations of toluene and their partners did not show a reduction in fecundity. In women (39 periods occurred during exposure) fecundity was reduced (FR 0.47, 95% confidence interval (95% CI) 0.29 to 0.77). Neither, restriction to only the first period nor exclusion of PUNPs changed the results (FR 0.48, 95% CI 0.24 to 0.97). CONCLUSION: After considering possible biases, low daily exposure to toluene in women seems to be associated with reduced fecundity. This result is in accordance with other findings for organic solvents and supports both the hypotheses that (a) organic solvents could affect hormonal regulation, and that (b) organic solvents increase early fetal losses which in turn contributes to longer times of unprotected intercourse. PMID- 10472315 TI - Role of individual susceptibility in risk assessment of pesticides. AB - OBJECTIVES: This study presents criteria for assessing the individual pesticide burden of workers in the chemical industry. METHODS: A group of 1003 workers exposed to methylparathion or ethylparathion (alkyl phosphates), propoxur (carbamate), or cyfluthrin (pyrethroid) was investigated. After exposure to methylparathion or ethylparathion the methylparathion or ethylparathion and methylparaoxon or ethylparaoxon concentrations in plasma, the p-nitrophenol concentration in urine, and the activities of cholinesterase and acetylcholinesterase were measured. For exposure to propoxur the propoxur concentration in plasma, the 2-isopropoxyphenol concentration in urine, and the cholinesterase and acetylcholinesterase activities were measured. For exposure to cyfluthrin the cyfluthrin concentration in plasma was measured. RESULTS: At the same propoxur concentration only workers with a low individual acetylcholinesterase activity reported symptoms. Workers who metabolised cyfluthrin rapidly reported less symptoms than workers with a lower rate of metabolism. This tendency was also evident in cases of mixed exposure (cyfluthrin and methylparathion). CONCLUSIONS: In the assessment of exposure to pesticides susceptibility of the individual person has to be considered. PMID- 10472316 TI - Occupational IgE sensitisation to phytase, a phosphatase derived from Aspergillus niger. AB - OBJECTIVE: Phytase is a phosphatase derived from Aspergillus niger that enhances phosphate bioavailability in the gut, and therefore has been increasingly used as an animal feed additive since the early 1990s. The aim of this study was to assess whether work related respiratory symptoms among workers in a so called premix factory producing animal feed additives, could be due to type I (mediated by immunoglobulin E (IgE) allergic sensitisation to phytase. METHODS: Preparations of specific IgE against phytase as used in the factory were assessed by enzyme immunoassay (EIA) in serum samples of 11 exposed workers who regularly handled the enzyme, in 11 office and laboratory workers of the same plant (non exposed internal controls), and in 19 laboratory animal workers as external controls. The factory workers also completed a questionnaire on common and work related respiratory symptoms. RESULTS: Depending on the cut off level in the EIA for IgE, and the preparation used as coated allergen, antiphytase sensitisation was found in one to four of the 19 external controls, in one to five of the 11 internal controls, and in four to 10 of the 11 exposed workers. Strongest IgE reactions were found in four exposed workers who reported work related respiratory symptoms, particularly wheezing, and in one internal control who possibly had become sensitised because the structure of the factory building did not preclude airborne exposure in the offices and corridors of the plant. Experiments with inhibition EIA for IgE showed that (a) phytase of another commercial source was only partially cross reactive with phytase as used in the premix factory, and (b) phytase used as an animal feed additive did not cross react with common mould extracts, except for extracts from the species of origin, Aspergillus niger. The amount of IgE binding phytase in Aspergillus niger was estimated to be between 0.1% and 1% of the extractable mould proteins. CONCLUSIONS: Phytase is a potentially important new occupational allergen causing specific IgE immune responses among exposed workers. Such IgE sensitisation could probably be the cause of work related asthmatic and other respiratory symptoms if no effective measures are taken to prevent airborne occupational exposure at sites where phytase is handled, particularly during addition of enzyme preparations to animal feed. PMID- 10472318 TI - Lack of combined effects of exposure and smoking on respiratory health in aluminium potroom workers. AB - OBJECTIVE: To investigate the combined influence on respiratory health of smoking and exposure in an aluminium potroom. METHODS: In a cross sectional study of 75 potroom workers (23 never smokers, 38 current smokers, 14 ex-smokers) and 56 controls in the same plant (watchmen, craftsmen, office workers, laboratory employees; 18 non-smokers, 21 current smokers, 17 ex-smokers), prevalences of respiratory symptoms and spirometric indices were compared. RESULTS: Smokers in the potroom group had a lower prevalence of respiratory symptoms than never smokers or ex-smokers, which was significant for wheezing (2.6% v 17.4% and 28.6% respectively, both p < 0.01), whereas respiratory symptoms in controls tended to be highest in smokers (NS). No effects of potroom work on the prevalence of respiratory symptoms could be detected. In potroom workers, impairment of lung function due to occupational exposure was found only in non-smokers, with lower results for forced vital capacity (FVC) (98.8% predicted), forced expiratory volume in one second (FEV1) (96.1% predicted) and peak expiratory flow (PEF) (80.2% predicted) compared with controls (114.2, 109.9, and 105.9% predicted; each p < 0.001). Conversely, effects of smoking on lung function were only detectable in non-exposed controls (current smokers v non-smokers: FVC 98.8% v 114.2% predicted; p < 0.01; FEV1 95.5 v 109.9% predicted; p < 0.05). CONCLUSIONS: In a cross sectional survey such as this, the effects of both smoking and occupational exposure on respiratory health may be masked in subjects with both risk factors. This is probably due to strong selection processes which result in least susceptible subjects continuing to smoke and working in an atmosphere with respiratory irritants. PMID- 10472317 TI - Improved method to measure urinary alkoxyacetic acids. AB - OBJECTIVES: To simplify the current preparation of samples, and to improve the specificity and reliability of the conventional analytical methods to measure urinary alkoxyacetic acids. METHODS: Samples containing alkoxyacetic acids including methoxy, ethoxy, and butoxyacetic acids (MAA, EAA, and BAA) were acidified with HCl and extracted with a mixed solvent of methylene chloride and isopropyl alcohol, then analysed by gas chromatography/mass spectrometry (GC/MS). RESULTS: Optimal results were obtained when pH was 1.05-1.45, the ratio of methylene chloride and isopropyl alcohol was 2:1, and when extraction time was 10 minutes. Over the concentration range 0.3-200 micrograms/ml, MAA, EAA, and BAA could be determined with a pooled coefficient of variation (nine concentrations, six replicate samples) of 5.55%, 6.37%, and 6.41%, respectively. Urine samples were stable for at least 5 months and 3 freeze-thaw cycles at -20 degrees C. The limits of detection of MAA, EAA, and BAA were 0.055, 0.183, and 0.009 microgram/ml, respectively. The matrix effect of urine samples was negligible for MAA and EAA, but were marginally significant for BAA. The average recoveries of alkoxyacetic acids were 99%-101%. In urine samples MAA from 15 exposed workers showed a strong linear correlation (r = 0.999, slope = 1.01) between the new GC/MS method and Sakai's GC method. CONCLUSIONS: The simplified non derivatisation pretreatment of samples coupled with GC/MS can provide a specific, sensitive, simple, safe, and reliable method for the biological monitoring of occupational exposure of ethylene glycol ethers. PMID- 10472319 TI - Compromised concentrations of ascorbate in fluid lining the respiratory tract in human subjects after exposure to ozone. AB - OBJECTIVES: Ozone (O3) imposes an oxidative burden on the lung in two ways. Firstly, directly as a consequence of its oxidising character during exposure, and secondly, indirectly by engendering inflammation. In this study the second pathway was considered by ascertaining the impact of O3 on the redox state of the fluid lining the respiratory tract 6 hours after challenge. METHODS: Nine subjects were exposed in a double blind crossover control trial to air and 200 ppb O3 for 2 hours with an intermittent exercise and rest protocol. Blood samples were obtained and lung function (forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1)) assessed before, immediately after, and 6 hours after exposure. Bronchoalveolar lavage (BAL) was performed 6 hours after challenge. Inflammation was assessed in BAL fluid (total and differential cell counts, plus myeloperoxidase concentrations), and plasma and BAL fluid redox state were determined by measuring concentrations of antioxidants and markers of oxidative damage. RESULTS: Neutrophil numbers in BAL fluid increased 2.2-fold (p = 0.07) 6 hours after exposure and this was accompanied by increased myeloperoxidase concentrations in BAL fluid (p = 0.08). On the other hand, BAL fluid macrophage and lymphocyte numbers decreased 2.5-fold (p = 0.08) and 3.1-fold (p = 0.08), respectively at this time. Of the antioxidants examined, only ascorbate in BAL fluid was affected by O3, falling in all subjects relative to air values (0.1 (0.0-0.3) v 0.3 (0.2-1.2) mumol/l (p = 0.008)). A marginal decrease in plasma ascorbate was also detected at this time (p < 0.05). Although the decrease in macrophage numbers seemed to be causally related to the increase in neutrophils (R = -0.79), myeloperoxidase concentrations (R = -0.93) and ascorbate concentrations (R = 0.6), no clear associations were apparent between ascorbate changes and neutrophils or myeloperoxidase concentration after O3. CONCLUSIONS: Ascorbate in the fluid lining the respiratory tract is depleted as a consequence of O3 exposure at 6 hours after exposure. This was contemporaneous with, although not quantitatively related to the increase in neutrophil numbers and myeloperoxidase concentrations. Decreased macrophage numbers 6 hours after O3 related to the degree of neutrophilic inflammation with populations conserved where ascorbate concentration in the fluid lining the respiratory tract were high after exposure. These results imply that ascorbate has a critical protective role against inflammatory oxidative stress induced by O3. PMID- 10472320 TI - Mass concentration and elemental composition of PM10 in classrooms. AB - OBJECTIVES: To investigate the sources of high concentrations of particles of < 10 microns diameter (PM10) in classrooms, observed in a previous study on childhood exposure to PM10, and to study the correlation between classroom and outdoor concentrations of mass and elements of PM10. METHODS: Measurements of PM10 were conducted in two schools and outdoors in Amsterdam, the Netherlands. Averaging time was 24 hours for the outdoor measurements and both 8 hours (school time) and 24 hours for the classroom measurements. Analysis by x ray fluorescence was used to measure the elemental composition of 55 samples from the 11 days when measurements were conducted simultaneously in both classrooms and outdoors. RESULTS: For most elements, classroom concentrations were considerably higher than outdoor concentrations, especially during school hours. The highest classroom/outdoor ratios were found for the elements from soils Si, Ca, and Ti. The only measured elements that were not increased were S, Br, Pb, and Cl, which are dominated by non-crustal sources. For S, Br, and Pb, which are generally associated with particles < 1 micron, significant correlations between classroom and outdoor concentrations and between the two classrooms were found. The other elements generally had low correlations. CONCLUSIONS: The results show that the high PM10 concentrations found in our classrooms are probably due to resuspension of coarse particles or suspension of soil material. Due to these excess coarse particles, the correlation between classroom and outdoor concentrations is lower for elements associated with coarse particles than for elements associated with fine particles. As the general composition of PM10 in classrooms differs from the composition of PM10 in ambient air, the high PM10 mass concentrations in classrooms can probably not be directly compared with ambient air quality guidelines. PMID- 10472321 TI - Relation between indicators for quality of occupational rehabilitation of employees with low back pain. AB - OBJECTIVES: To assess if the implementation of guidelines for occupational rehabilitation of patients with low back pain by means of process variables--a set of objective criteria for technical performance and continuity of care--led to a better outcome in clinical and return to work variables. METHODS: The study group consisted of 59 patients with at least 10 days of sick leave because of low back pain. Univariate analyses as well as multiple logistic regression and Cox's regression analyses were performed to assess the relation between quality of care and outcome. RESULTS: Process indicators for technical competence, continuity of care, and total performance were all significantly related to satisfaction of employees. Continuity of care and total performance were significantly related to working status at 3 months, and time to return to work. None of the process indicators was related to pain or disability after 3 months follow up. Satisfaction was not related to any of the other outcome variables. This indicates that if guidelines for occupational rehabilitation are met, outcome is better. CONCLUSION: Quality of the process of care was related to outcome. Interventions of occupational physicians need improvement in the areas of continuity of care and communication with treating physicians. The effectiveness of an improved intervention should be studied in a subsequent randomised clinical trial. PMID- 10472322 TI - Shoulder impingement syndrome in relation to shoulder intensive work. AB - OBJECTIVES: To analyse the risk of shoulder impingement syndrome relative to shoulder intensive work. METHODS: A cross sectional study of a historical cohort of 1591 workers employed between 1986 and 1993 at a slaughterhouse or a chemical factory. Workers not doing tasks in slaughtering or meat processing constituted the reference group. Intensity of shoulder work in meat processing tasks was assessed by video based observations. Information on shoulder disorders was collected by questionnaire and by physical examinations. Impingement syndrome was diagnosed when shoulder symptoms had been present for at least 3 months during the past year and there were signs of subacromial impingement in the corresponding shoulder at physical examination. Shoulder function was assessed at the same occasion with the Constant scoring technique. Prevalence of shoulder impingement syndrome was analysed according to job title and cumulative exposure. RESULTS: Prevalence ratio for shoulder impingement syndrome was 5.27 (95% confidence interval (95% CI), 2.09 to 12.26) among currently working and 7.90 (95% CI, 2.94 to 21.18) among former slaughterhouse workers. Transformed model based prevalence ratios according to years in slaughterhouse work showed an overall association between cumulative exposure and risk for shoulder impingement syndrome. CONCLUSIONS: This study supports the hypothesis that shoulder intensive work is a risk factor for impingement syndrome of the shoulder. Despite the historical cohort design healthy worker selection may have influenced the exposure-response relation found. PMID- 10472323 TI - Feasibility of cohort studies in Estonia. AB - OBJECTIVES: To describe the methodology and feasibility of follow up for vital status in retrospective cohort studies in Estonia. METHODS: A cohort of 7412 workers who had been employed at two factories in Tallinn between 1946 and 1988 was followed up for vital status from the date of first employment until death, emigration, or the end of the study, 31 December 1995, whichever occurred first. The cohort was electronically linked with the National Population Registry of Estonia that was created in 1992 and includes personal identification numbers of Estonian citizens and residents, and the Mortality Database that contains information from death certificates issued in 1983-95. A manual search was carried out on several non-computerised population data sources and archives. RESULTS: By 31 December 1995, the vital status of 6780 (91.5%) subjects could be traced (4495 (60.6%) subjects were alive, 1993 (26.9%) had died, and 292 (3.9%) had emigrated). Analysis by calendar period of leaving work showed that the proportion of subjects traced was lowest in the group of workers who had left work between 1946 and 1955 (58.4%), especially those whose age at leaving work was < 30 (53.2%) or > 60 years (42.3%). Among subjects who left work in 1956-65, 1966-75, and 1976-88, the follow up rate was 84.7%, 94.6%, and 98.2%, respectively. CONCLUSIONS: The findings, which are especially important for occupational epidemiology, confirm the feasibility of conducting retrospective cohort studies in Estonia. Most of the issues discussed in the paper apply to other former Soviet countries. PMID- 10472324 TI - Mitomycin C for metastatic prostate cancer: final analysis of a randomized trial. AB - BACKGROUND: Almost all patients that undergo hormonal manipulation for metastatic prostate cancer will ultimately progress because of hormone resistance. Therefore we assessed the effect of early addition of intravenous Mitomycin C to orchiectomy in patients with newly diagnosed metastatic prostate cancer. PATIENTS AND METHODS: 178 patients with histologically proven and previously untreated metastatic prostate cancer were included in a prospective, randomized multicenter trial. Randomization was done centrally between orchiectomy alone and orchiectomy with Mitomycin C. 148 patients were evaluable. RESULTS: At the final analysis 139 patients have deceased. The remaining 9 patients are still alive, but all present progression. There was no statistically significant difference in the real time to progression, or in the estimated cancer related and overall survival between both groups. Mean time to progression was 29 months in group 1 (orchiectomy alone), and 26 months in group 2 (orchiectomy and Mitomycin C) (p = 0.64). Mean time to cancer related death was 32 months and mean overall survival was 31 months in both groups. CONCLUSIONS: Mitomycin C has no beneficial effect when used in conjunction to orchiectomy in patients with newly diagnosed metastatic prostate cancer. PMID- 10472325 TI - Thymidine kinase (TK) gene therapy of solid tumors: valacyclovir facilitates outpatient treatment. AB - BACKGROUND: In a Phase I study replication-deficient adenovirus containing the herpes simplex virus (HSV) thymidine kinase (TK) gene (AdV-HSV-TK) was instilled intraperitoneally in patients with recurrent ovarian cancer. Patients were treated with Acyclovir (ACV) or Valacyclovir (VCV) as enzymatic substrates. The purpose of this study was to compare serum levels of ACV and VCV. PATIENTS AND METHODS: The antiherpetic prodrug and Topotecan (1.0 mg/m2 over 30 minutes each day for 5 days) were started 24 hours after vector application. Eight patients received ACV (15 mg/kg i.v. over one hour every 8 hours for 42 doses), two patients were started on ACV for 5 days and then switched to oral VCV (2 g every 8 hours for a total of 42 doses). Blood samples were obtained 20 minutes prior to each drug. RESULTS: Serum levels of ACV and VCV (converted to ACV) were comparable. CONCLUSIONS: Suicide gene therapy with TK is under investigation in a variety of solid tumors. Replacing ACV by VCV will offer a cost-effective alternative and will significantly reduce duration of hospital stay improving quality of life and facilitating an outpatient gene therapy concept. PMID- 10472326 TI - Change in telomerase activity during human colorectal carcinogenesis. AB - Telomerase activities in endoscopically resected colorectal adenomas, surgically resected colorectal cancers and adjacent normal colonic mucosa were examined semiquantitatively by a polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) assay. All normal mucosa (n = 15) presented weak telomerase activity (mean +/- SE: 0.99 +/- 0.00). When the value of 1.00 was arbitrary given to the mean activity of normal mucosa, the telomerase activity in the adenomas (n = 14) was up-regulated (2.01 +/- 0.22) relative to the normal mucosa. The telomerase activity in the high-grade atypia (severe atypia and carcinoma in situ) (2.58 +/- 0.34) was significantly higher than that in the low grade (mild and moderate) atypia (1.59 +/- 0.18) (P < 0.05), and the adenomas 10 mm or more in diameter presented significantly higher telomerase activity (2.56 +/- 0.09) than compared to the smaller ones (1.44 +/- 0.17) (p < 0.05). Carcinomas (n = 20), showed a telomerase activity that varied from 0.97 to 16.93, which was than the greater mean telomerase activity (6.96 +/- 1.25) noted in the adenomas. The telomerase activity in the carcinomas tended to be higher in the larger (> or = 4 cm), histologically less-differentiated (moderately differentiated), late-stage (Dukes C + D), and nodal metastatic tumors, suggestive of unfavorable prognosis. These results suggests that the weak telomerase activity in normal colonic mucosa is gradually activated during the course of colorectal carcinogenesis. PMID- 10472328 TI - Comparison of carcinoembryonic antigen in tissue and serum with grade and stage of colon cancer. AB - BACKGROUND: Serial plasma carcinoembryonic antigen (CEA) levels are routinely used to detect postoperative recurrence of colon cancers. However, levels of intrinsic CEA production by individual tumors, which might be helpful for interpreting serum CEA levels, are not routinely available. MATERIALS AND METHODS: CEA levels were determined in twenty tissue biopsy specimens of colon carcinoma and sixteen normal colonic tissue specimens from India, by immunohistochemistry (IH) and by quantitative enzyme immunoassay (EIA). Serum samples from 62 colon cancer patients and 25 healthy blood donors were analyzed for CEA by EIA. RESULTS: Tissue CEA levels were highest for well differentiated adenocarcinomas (5.2-37 micrograms/g protein) with progressively lower levels seen in moderately differentiated and poorly differentiated tumors, and in normals. The intensity of immunostaining paralleled the levels determined quantitatively. Mean serum CEA levels were 1.5 ng/ml for normals and 4.2, 6.4, 23, and 102 ng/ml for Dukes' A, B, C and D stage tumors, respectively. CONCLUSION: Preoperative serum CEA levels do not, in themselves, take into account differences in CEA production between individual tumors. Determination of tumor CEA content by quantitative or immunohistochemical methods, could be a useful adjunct for the clinical management of colon carcinoma, by improving interpretation of serum CEA levels. PMID- 10472327 TI - Cancer risk associated with simian virus 40 contaminated polio vaccine. AB - BACKGROUND: The presence of SV40 in monkey cell cultures used in the preparation of the polio vaccine from 1955 through 1961 is well documented. Investigations have consistently demonstrated the oncogenic behavior of SV40 in animal models. Early epidemiologic studies were inadequate in demonstrating an increase in cancer incidence associated with contaminated vaccine. Recently, investigators have provided persuasive evidence that SV40 is present in human ependymomas, choroid plexus tumors, bone tumors, and mesotheliomas, however, the etiologic role of the virus in tumorigenesis has not been established. MATERIALS AND METHODS: Using data from SEER, we analyzed the incidence of brain tumors, bone tumors, and mesotheliomas from 1973-1993 and the possible relationship of these tumors with the administration of the SV40 contaminated vaccine. RESULTS: Our analysis indicates increased rates of ependymomas (37%), osteogenic sarcomas (26%), other bone tumors (34%) and mesothelioma (90%) among those in the exposed as compared to the unexposed birth cohort. CONCLUSIONS: These data suggest that there may be an increased incidence of certain cancers among the 98 million persons exposed to contaminated polio vaccine in the U.S.; further investigations are clearly justified. PMID- 10472329 TI - Modified true-color computer-assisted image analysis versus subjective scoring of estrogen receptor expression in breast cancer: a comparison. AB - BACKGROUND: Hormone receptor expression can be quantified by computerized image analysis in immunohistochemically stained specimens. When comparing semiquantitative scoring with computerized image analysis a review of the literature shows contradictory findings concerning the correlation of these two methods. Recent technical approaches have been developed with true-color computer assisted image analysis facilitating new measurement designs. We performed a study with a new approach using the principle of semiquantitative assessment of hormone receptor content and measuring two different binary images (immunohistochemically stained nuclear area and total nuclear area). MATERIAL AND METHODS: Eighty formalin-fixed, paraffin-embedded and immunohistochemically stained breast cancer specimens were assessed for estrogen receptor expression by true color computer-assisted image analysis and by conventional light microscopy scoring according to Remmele (immunoreactive score (IRS) = staining intensity (SI) x percentage of positive cells (PP)). The results of both methods were correlated. RESULTS: Mean optical density (MOD) and subjective scoring of SI as well as stained nuclear area vs. total nuclear area and subjective scoring of stained cells (PP) showed a high correlation (Spearman correlation coefficient: 0.95, p-value: 0.0001 and 0.64, p-value: 0.0001, respectively). CONCLUSION: On the basis of this new technical approach our results confirm the correlation of semiquantitative hormone receptor scoring and quantitative computer-assisted image analysis. We believe that by automating electronic analysis in the near future we will be able to establish reliable observer-independent evaluation of immunohistochemical variables ensuing comparability in multi-center trials and cost efficiency. PMID- 10472330 TI - Cancer protection in xeroderma pigmentosum variant (XP-V). AB - We describe herein a brother and sister diagnosed with xeroderma pigmentosum variant (XP-V) in early adult life, who presented with increased sensitivity to sunlight and with cutaneous carcinomas on sun-damaged skin. The 27-year-old male farmer (Case 1.) was diagnosed with advanced squamous cell carcinoma (SCC) and multiple actinic lesions. Surgical removal of these lesions was performed. Three months later he died of multiple pelvic metastases of SCC. His 29-year-old sister (Case 2.) was operated on for different tumors, histologically SCC-s or basal cell carcinomas (BCC), or praecancerous conditions many times. After a two year interval she was treated with low dose isotretinoin (2 mg/body weight). Diagnosis of XP-V was based on unscheduled DNA analysis (USD) and on clinical symptoms. We observed that during the long lasting isotretinoin treatment the tumor frequency dropped to a quarter. Therefore, the isotretinoin treatment seems to be a good approach for cancer prevention in conditions with high predisposition to skin cancer, such as in XP-V. PMID- 10472331 TI - Diagnostic value of anti-alpha FP antibody levels in a metastatic germ cell tumor of unknown primary site. AB - BACKGROUND: A 21 year old man with a metastatic germ cell tumor of unknown primary not responding to chemotherapy was scheduled to have a blind bilateral orchiectomy to eradicate the possible primary site although palpation and ultrasonography of the testicles had always been normal. METHOD: The patient underwent a radioimmunoscintigraphy with Anti-alpha FP antibody scan (AFP-Scan). RESULTS: On the basis of the scintigraphic results the patient underwent a left orchiectomy and additionally removal of the lymph node metastases. Histology revealed the presence of an in situ carcinoma in the left testis and a mixed tumor present in the abdominal lymph node metastases. Fluorescent in situ hybridization on tumor cells did not show any abnormalities related to chromosome 12, a finding connected with the somatic type of germ cell tumors. CONCLUSION: Anti-alpha FP antibody scan was helpful in detecting the primary site and saving the life of the patient without resulting in hypogonadism. PMID- 10472332 TI - Expression and clinical relevance of NY-ESO-1, MAGE-1 and MAGE-3 in neuroblastoma. AB - Human genes NY-ESO-1, MAGE-1 and MAGE-3 code for antigens which are expressed in malignancies of various histological types but not in normal tissues except testis. These antigens might therefore represent potential targets for specific immunotherapy. We studied the expression of genes NY-ESO-1, MAGE-1 and MAGE-3 in 98 neuroblastoma tumors by reverse transcription-polymerase chain reaction (RT PCR). MAGE-1 was expressed in 66%, NY-ESO-1 in 36% and MAGE-3 in 33% of the tumors. NY-ESO-1 gene expression was associated with age older than one year (p = 0.017), more differentiated tumor histology (p = 0.044), elevated urinary vanillylmandelic acid (VMA, p = 0.018) and normal serum ferritin levels (p = 0.023). MAGE-1 expression correlated significantly with normal serum ferritin levels (p = 0.009) and absence of MycN amplification (p = 0.007) while MAGE-3 expression was associated with absence of metastasis (p = 0.027). We conclude that approximately 70% of the neuroblastoma tumors express at least one of the genes coding for NY-ESO-1, MAGE-1 or -3, respectively. PMID- 10472333 TI - Study of minimal residual disease in B-cell lineage acute lymphoblastic leukemia of childhood, using clone-specific probes. AB - In B-cell lineage acute lymphoblastic leukemia (B-ALL), the clonal rearrangements of the immunoglobulin heavy chain gene locus (IgH), can be used as a molecular marker for the detection of minimal residual disease (MRD). Patients in complete remission may still harbor leukemic cells undetectable by conventional methods such as light-microscopic examination, immunophenotyping and cytogenetics. 30 children with B-ALL were screened at diagnosis by polymerase chain reaction (PCR) for their IgH gene repertoire. 7/30 patients were extensively studied using patient-specific oligonucleotide probes derived from the sequence analysis of bone marrow (BM) samples at diagnosis. 210 PCR products from follow-up BM samples corresponding to these 7 patients were hybridized with the appropriate clone specific probe in order to detect MRD with high sensitivity and specificity. All the patients were in morphological remission during and after therapy. 25/30 patients were PCR positive at diagnosis. 4/7 patients who were examined for MRD had detectable disease in various periods after diagnosis. Molecular signs of residual cells can persist for a long time during and after therapy. Long term follow-up of MRD could determine the period of therapy and predict relapse, indicating therapeutic interventions. PMID- 10472334 TI - Pleomorphic T-cell lymphoma with chondropathia tuberosa; a case report and review. AB - Pleomorphic T-cell-lymphoma (anaplastic IgA-plasma cell tumor) belongs to the group of malignant non-Hodgkin's lymphomas (NHL). The histological and immunophenotypical subtypes differ with genetic and environmental etiologic factors. Lymphomas arise from the clonal proliferation of precursor cells within lymphoid organs with acquired chromosomal abnormalities. Approximately 5% of all primary malignant bone tumors are NHL, the majority of diffuse large B-cell type. Our case history can be regarded as the first published in English language reporting on a pleomorphic T-cell-lymphoma imitating a Chondropathia tuberosa (Tietze's syndrome): The tumor appeared with a tender tumescence over the sternum and a painful swollen left sterno-clavicular joint--as a rule a typical sign for Tietze's syndrome. Only sternal puncture followed by immune histology confirmed an anaplastic IgA-plasma cell tumor. The primary tumor, and later on an osteolysis of cervical vertebrae I-III with a complete destruction of the axis and an affection of the dens atlantis could not be detected by radiographic examinations. Moreover, this tumor infestation could only be depicted by the magnet resonance imaging and the computed tomography. Furthermore, other results of our report are the very rare manifestation of a T-cell lymphoma as a pedicled tumor near the pancreas or of pancreatic origin and the excellent result of an autologous stem cell transplantation. Reviewing the literature, we want to discuss the present scientific and clinical standards of diagnosis, progress and treatment of Chondropathia tuberosa and T-cell lymphoma, and we want to point out some new aspects of both diseases. PMID- 10472335 TI - Individual 5FU-dose adaptation schedule using bimonthly pharmacokinetically modulated LV5FU2 regimen: a feasibility study in patients with advanced colorectal cancer. AB - AIM OF THE STUDY: SFU-dose adaptation optimal schedule using bimonthly LV5FU2 regimen was modulated by previously validated individual pharmacokinetic parameters, in 38 patients with advanced colorectal cancer. METHODS: At the 1st cure, 5F-U pharmacokinetic parameters (particularly the area under curve (AUC) in mg.h/l.m2) were calculated in all patients. In 19 patients (A), 5FU infusion doses were progressively increased from 25 to 50% (at every cycle), according to AUC levels from 2nd to 6th; in 19 consecutive patients (B), 5FU infusion doses were increased, at the same time, at the 2nd cure, according to a protocol taking in account AUC at the 1st cure: increase of 150% it AUC < 5, of 100% if 5 < AUC < 10, of 50% if 10 < AUC < 15, of 25% if 15 < AUC < 20 in case of toxicity < grade 3. RESULTS: a) AUC in all patients, at the beginning of the treatment averaged 9.05 +/- 3.115 (range from 3.9 to 16.41). Large interindividual variability was observed. b) FU infusion doses at the 2nd cure, increased 40% in group A and 82% in group B. Corresponding AUC increased respectively of 42% and 96%. 3-WHO toxicity 23 (per cycle) occurred not very frequently (8% haematological, 6% digestive and 2% cutaneous toxicity) and remained acceptable. CONCLUSION: This feasibility study established a 5FU-dose intensification optimal strategy within the bimonthly LV5FU2 schedule with a control for the risk of toxicity. This study constitutes the basis for a multicentre phase II trial to evaluate the importance of this approach in terms of efficacy and survival. PMID- 10472336 TI - Ep-CAM--a marker for the detection of disseminated tumor cells in patients suffering from SCCHN. AB - Disseminated tumor cells are considered as the origin of metastases. Since the number of circulating tumor cells in the bone marrow or the peripheral blood of patients correlates well with the tumor stage, their early detection is an important feature for the identification of high risk patients. We therefore investigated the pan-carcinoma antigen Ep-CAM for its suitability to serve as a specific marker for disseminated tumor cells in patient with squamous cell carcinoma of the head and neck (SCCHN). In order to detect small numbers of tumor cells in early tumor stages, we developed and describe here a RT-PCR assay that detects a single tumor cell within 10(5) normal cells. We examined bone marrows from patients and healthy donors and demonstrate that Ep-CAM can be used as a tumor marker for the diagnosis of single tumor cells in patients with SCCHN. PMID- 10472337 TI - Endothelins and their receptors in cirrhotic and neoplastic livers of Canadian and Chinese populations. AB - BACKGROUND: Endothelins (ETs) are 21 amino acid peptides with widespread tissue distribution and functions. In this study, we retrospectively investigated immunoreactive ET-1, ET-3 as well as ET receptors by ligand binding and autoradiography in hepatic cirrhosis and neoplasms. MATERIALS AND METHODS: Formalin fixed paraffin embedded tissues from 30 hepatocellular carcinomas (HCC), 4 fibrolamellar carcinomas (FLC), and 7 liver metastatic adenocarcinomas (Ad) from colon were collected from the Pathology Department of London Health Science Centre. Adjacent cirrhotic livers were obtained from 17 cases and adjacent normal liver was present in 12 cases. In addition, 15 HCCs, 6 cirrhotic and 8 normal livers were obtained from Normal Bethune University for Medical Sciences in China. The slides were stained for ET-1 and ET-3 with a polyclonal antibody and scored. Autoradiographic localization of ET-receptors with 125I-ET-1 was carried out in some of the cases. RESULTS: In the normal liver, hepatocytes, biliary epithelium, vascular endothelium and smooth muscle cells were positive for both ET-1 and ET-3. Higher immnunoreactivity for ET-1 and ET-3 was seen in cirrhosis. HCCs showed variation in immunoreactivity, with overall scoring not different from normal livers. FLCs showed consistent higher immunoreactivity for both ET-1 and ET-3, while in Ads the immunoreactivity was decreased. Increased ET receptors, representing both ETA and ETB subtypes were seen in both cirrhosis and in HCC. CONCLUSION: Alterations in both ETs and their receptors were found in cirrhosis and neoplastic liver diseases. PMID- 10472338 TI - Biological characterization of central and peripheral primary non small cell lung cancers (NSCLC). AB - BACKGROUND: Non Small Cell Lung Carcinomas (NSCLC) comprise 90% of all lung carcinomas. Studies have demonstrated a preferential central (bronchus-derived) localization for squamous cells, whereas adenocarcinomas are frequently peripheral (bronchiolo-alveolus derived). It has been suggested that exposure to carcinogenic insults including cigarette smoke, may induce different types of tumors in different locations. MATERIALS AND METHODS: Forty one NSCLC patients staged according to WHO and TNM were considered for localization and biological parameters (p53 expression, cell ploidy and S-phase). RESULTS: p53 overexpression was found more frequently in central than in peripheral tumors (69% vs 39%) (p = 0.074). Central tumors were more aneuploid (69%) than peripheral ones (46%) (p = 0.03) No difference in smoking habit was observed in the two groups. CONCLUSIONS: Our results suggest that there is no apparent biological difference between these two groups of NSCLCs, and that the smoking does not play a role in either histotype determination or biological behavior. PMID- 10472339 TI - Combination of cisplatin, epirubicin, and cyclophosphamide (PEC regimen) in advanced or recurrent endometrial cancer: a retrospective clinical study. AB - The combination of cisplatin (50 mg/m2), epirubicin (60 mg/m2), and cyclophosphamide (600 mg/m2) (PEC regimen) was given every 4 weeks to 19 patients with advanced (n.2) or recurrent (n.17) endometrial cancer. The median number of cycles delivered to each patient was 5 (range, 2-8). All patients were evaluable for toxicity and 16 for response. Two (12.5%) patients experienced a complete response and 5 (31.2%) had a partial response, for an overall objective response rate of 43.7%. The median duration of objective response was 10 months (range, 3 28 months). Median survival was 10 months (range, 3-68+ months) in the whole series. According to response to chemotherapy, median survival was 12 months (range, 3-68+ months) for responders and 9 months (range, 6-17 months) for nonresponders. Hematologic toxicity was relatively frequent but it could be easily managed, and significant nonhematologic toxicities were not found except for nausea and vomiting. In conclusion, PEC regimen has a good activity in advanced or recurrent endometrial cancer, but the short duration of responses limits the impact of the treatment on survival time. PMID- 10472340 TI - Vascular endothelial growth factor expression is an independent negative predictor in extrahepatic biliary tract carcinomas. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is one of the most important factors for angiogenesis in various tumors. However, its clinical significance in extrahepatic biliary tract carcinoma remains unclear. MATERIALS AND METHODS: To clarify the association of VEGF expression with the clinicopathological features of the disease, surgical specimens from 51 patients were examined for VEGF expression by immunohistochemistry. RESULTS: Sixteen of 51 (31.4%) cases were positive for VEGF. VEGF immunoreactivity showed a tendency toward association with lymph node metastasis, M category and peritoneal recurrence (p = 0.059, 0.086 and 0.069, respectively). Overall survival in patients with VEGF positivity was significantly worse than in those with negativity (p = 0.033 by log-rank test). Multivariate Cox regression analysis revealed that VEGF expression was an independent negative predictor (p = 0.025). CONCLUSION: VEGF immunoreactivity is an independent negative predictor in extrahepatic biliary tract carcinomas. PMID- 10472342 TI - Efficacy of treatment of colon, lung and breast human carcinoma xenografts with: doxorubicin, cisplatin, irinotecan or topotecan. AB - Given that human cancer xenografts tend to retain chemosensitivities similar to the cancerous tissue of origin, human carcinoma xenografts grown in nude mice were tested for sensitivity to four drug protocols: doxorubicin at 5 mg/kg, i.v., q5d; irinotecan at 60 mg/kg, i.v., q4d; cisplatin 5 mg/kg, i.p., q7d; and topotecan 1.5 mg/kg, p.o., qd (5 of 7 days). Irinotecan and doxorubicin protocols either halted or caused significant regression of the breast cancer cell lines (MCF7, MDA-MB 231 and T47D). None of the protocols tested resulted in significant regression in the lung cancer xenografts (H460, A549 and H226) although both irinotecan and doxorubicin did halt growth of the H226 xenograft. The ability of the irinotecan treatment to cause regression of xenograft size in all three colon cancer cell lines (SW620, COLO205 and HT29) justifies further clinical trials of irinotecan as an especially promising drug for the treatment of colon cancer. PMID- 10472341 TI - Premenopausal breast cancer patients treated with a gonadotropin-releasing hormone analog alone or in combination with an aromatase inhibitor: a comparative endocrine study. AB - BACKGROUND: The combination of a GnRH analogue and an aromatase inhibitor can induce a complete estrogen blockade in premenopausal breast cancer patients. MATERIAL AND METHODS: Twenty-one premenopausal women with advanced breast cancer were randomised to receive the GnRH analog triptorelin (3.75 mg i.m. monthly; n = 10) alone or in combination with the aromatase inhibitor formestane (4-OHA, 500 mg i.m. fortnightly; n = 11) to compare the effect of both treatments on the patients' estrogenic milieu. Therefore, serum estrogen, gonadotropin and sex hormone-binding globulin (SHBG) levels were investigated before the start of treatment and subsequently over a three-month period. RESULTS: There was a significant between-group difference in estrogen suppression during therapy. In comparison with baseline values, after four weeks of treatment the estradiol levels decreased by an average of 86.9% (95% CI, 70.5-94.2%) in the group treated with triptorelin alone and by 97.3% (95% CI, 94.1-98.8%; P = 0.0422) in the combination group; the respective figures for estrone were 48.5% (95% CI, 27.5 63.5%) and 70.4% (95% CI, 52.3-81.6%; P = 0.0007) and for estrone sulfate 56.7% (95% CI, 40-68.8%) and 80.5% (95% CI, 69.4-87.6%; P = 0.0055). No difference was observed between the groups in terms of gonadotropin suppression; both treatment modalities led to a slight but delayed decrease in SHBG levels. Three of the patients treated with triptorelin alone experienced tumor regression compared with four patients in the combination group. No appreciable side effects of the combination therapy were observed. CONCLUSION: The treatment of premenopausal patients with triptorelin plus 4-OHA is feasible and leads to a much greater inhibition of main circulating estrogens than treatment with the analog alone. Since the combination of a GnRH analog and an aromatase inhibitor might potentially enhance the anti-tumor efficacy of the analog alone owing to more favorable endocrine effects, such a therapeutic approach deserves more extensive evaluation in the clinical setting. PMID- 10472343 TI - Polyamines in human breast cancer and its relations to classical prognostic features: clinical implications. AB - Experimental evidence suggest an important role of polyamines in breast cancer development. Polyamines have been determined in tissue and erythrocyte samples from 100 patients with primary invasive breast cancer and 30 patients with fibroadenomas. Statistical analysis was performed in order to determine the prognostic value of the polyamine patterns of tumor tissues and erythrocytes in comparison with clinical and histological prognostic factors. In malignant tissues, polyamine levels were significantly higher than in benign tissues. They correlated with markers of tumor aggressivity (axillary node involvement and especially with markers of high mitotic rate as Ki-67 staining, histological grade). No correlation was found between estrogen and progesterone status, tumor size and polyamine concentrations. Erythrocyte polyamines levels were identical between cancer patients and controls. The knowledge of the polyamine pattern in breast cancer could become useful in clinical practice particularly if polyamine metabolism is targeted as a therapeutic approach. PMID- 10472344 TI - A new administration schedule for amifostine as a radioprotector in cancer therapy. AB - Thus far, amifostine, a new radioprotective substance, had been given as short infusion 30 minutes before radiotherapy. In our investigation for the first time we have administered the substance as bolus injection (200 mg/m2 dissolved in 10 ml sodium chloride 0.9%). 42 patients with malignancies of the head and neck region, recurrences of rectal carcinomas and bronchial carcinomas had been treated with different solution volumes over different time periods. Using amifostine as a bolus injection had led to a statistically significant decrease in acute toxicity (p = 0.012). In comparison to short infusion (15 minutes) we have documented the same radioprotective impact. Hence, bolus injection of amifostine is feasible and can spare manpower and time in the department of radiotherapy. PMID- 10472345 TI - DNA ploidy alterations detected during dedifferentiation of periosteal chondrosarcoma. AB - DNA ploidy of a case with dedifferentiated periosteal chondrosarcoma was analyzed by DNA cytofluorometry. The diagnosis of primary periosteal chondrosarcoma was made on the basis of the radiographic and histological findings. At 4 years after marginal resection, the tumor recurred locally and metastasized to various organs. The patient died of disease 2 years later. Histologically, there were two components, chondrosarcoma and malignant fibrous histiocytoma, in the recurrent and metastatic tumors. DNA ploidy analysis of multiple samples revealed that the primary lesion was composed of many diploid cells with some tetraploid and octaploid cells, whereas in the dedifferentiated area, there were many aneuploid cells which were not recognized in any area of the primary tumors. This case illustrated that DNA ploidy alteration of euploidy to aneuploidy is closely correlated with the process of dedifferentiation in chondrosarcoma. PMID- 10472346 TI - Treatment of refractory metastatic breast cancer with 5-fluorouracil and levofolinic acid as 48 hours continuous venous infusion. AB - Twenty-four consecutive patients with metastatic breast carcinoma (MBC) refractory to first line chemotherapy were treated with high-dose folinic acid (FA) 100 mg/m2 diluted in 250 cc of normal saline as 2 hour infusion followed by 5-fluorouracil (5FU) 400 mg/m2 bolus then 5FU 600 mg/m2 as continuous infusion for 22 hours. This therapy was repeated for 2 consecutive days. Chemotherapy was repeated every 15 days. All enrolled patients were evaluable for objective response. A complete response was achieved in 1 patient (4%) and a partial response in 6 cases (25%) for an overall response rate of 29% (confidence limits 18%-39%). The median duration of objective responses was 8.4+ months (range 3.0+/12.8). Six patients showed no change (25%) with a median duration of 4.0 months 11 patients progressed (46%). A subjective improvement in tumor-related symptoms was reported by all responding patients and in 3 patients with no change. Most patients (7/10) with symptomatic bone lesions had a subjective improvement with reduction in analgesic drugs consumption. Objective responses were observed at all sites of disease. In fact, responses were seen in the skin liver lung bore and rodal metastases. The median overall survival was 13.0+ months (range 4.0/16.2+). Over a total of 160 cycles (a mean of 6.6 cycles/patient) grade 1-2 leukopenia was seen in 9 patients (37%) grade 1 thrombocytopenia in 4 patients (17%) and grade 1 anemia in only 2 cases (8%). Grade 3-4 leukopenia or thrombocytopenia were not seen. Phlebitis at the injection vein occurred in 3/10 patients (30%) which refused to implant a central line. In patients with a central line or a port-a-cath no cases of vascular, toxicity were seen. Gastrointestinal toxicity was very mild with 9 patients (37%) suffering from grade 1-2 nausea/vomiting 6 patients (25%) complaining of grade 1 2 diarrhea and 6 patients with grade 1-2 stomatitis. Hand-foot syndrome was observed in only 1 patient. No cases of grade 3-4 gastrointestinal toxicities have been, observed. No cases of cardiotoxicity and/or neurotoxicity were recorded. The combination of high-dose FA and 5FU given as 48 hour continuous venous infusion every 2 weeks is active, at least in terms of objective response rate and tumor-related symptoms palliation against anthracycline-refractory MBC. These results compare favorably with bolus administration of FA and 5FU or other salvage regimens. PMID- 10472347 TI - The organization of a counselling service for the families of patients undergoing bone marrow transplant. AB - The organisation of our Counselling Service for the relatives of patients undergoing bone marrow transplant is structured in various phases. During the first phase, contact is established with the relatives. The therapist introduces himself to the relatives when the patient is hospitalized and enters the sterile ward. The second phase consists in an illustration of the objectives and purpose of the Service. During this phase, two weekly interviews lasting approximately one hour are proposed, plus assessment tests (CFI, Camberwell Family Interview- and MMPI--Minnesota Multiphasic Personality Inventory). A third phase, which is often contextual to the presentation of the Service, consists in the recognition of the needs and suffering of the given relative. Here, the therapist acts as a receptacle for the anxieties and fear of death brought to him by the subject, thus empathetically accepting these states of mind and legitimizing them. Moreover, our model of intervention envisages the formation of support groups for the relatives of patients suffering from blood cancers. A further phase concerns the exchange of information between the two therapists helping, respectively, the relative and the patient, and also with medical and nursing staff. To further the same aims, for a number of years now, our Service has been organising intervention groups for nursing staff working at CTMO. The final phase is counselling, which takes place twice a week. There are two fundamental areas of intervention that must be taken into account for families of subjects suffering from blood cancers: providing information and offering support. From our experience, it emerges that the relatives need to obtain information on organic aspects of the illness and the transplant. PMID- 10472348 TI - Correlation between uptake of Tc-99m sestaMIBI and prognostic factors of breast cancer. AB - The purpose of this retrospective study was assessment of correlation between Tc 99m sestaMIBI uptake and some prognostic factors of breast cancer. The following prognostic factors have been included in this study: size of the tumour, age of the patients, axilla node involvement, oestrogen and progesterone receptor (ER, PR) status, grading system of Bloom-Richardson and Ki-67 antigen expression. METHODS: 79 patients were enrolled in this study, with 85 lesions confirmed as primary breast cancers. Mean age of patients was 53 years. Scintimammography (SMM) was performed after intravenous injection of 740MBq. At 5-10 min after injection standard planar images were obtained in prone lateral and anterior supine views. Assessment of correlation between known prognostic factors of breast cancer and uptake of MIBI (evaluated as a tumour to background ratio-TBR) was performed used non-parametric (Kendall-tau correlation) statistical analysis. RESULTS: There were 85 breast cancers (73 invasive ductal carcinomas, 11 DCIS (ductal carcinoma in situ) and 1 lobular carcinoma. There was positive correlation between TBR Tc-99m MIBI uptake and size of the tumour (t = 0.19, p = 0.01), presence of axilla node involvement (t = 0.2, p = 0.006) and also grade of the IDC tumours evaluated using Bloom-Richardson's criteria (t = 0.18, 0.03). There were negative correlation between TBR and presence of PR (t = -0.16, p = 0.02) and borderline negative correlation between TBR and age of patients (t = 0.137, p = 0.06) and also TBR and ER status (t = -0.135, p = 0.065). Patients who are younger and/or have PR or ER negative cancers have higher Tc-99m MIBI uptake. Patients who presented with high grade of malignancy (B-R) also have higher uptake of radiotracer. Also those with higher uptake of radiotracer often had axillary node involvement. This would suggest that more aggressive tumours have higher uptake of Tc-99m MIBI. Finally this study suggest correlation between in vivo uptake of MIBI and some of the known prognostic factors of breast cancer. PMID- 10472349 TI - Combined irinotecan, docetaxel and conventionally fractionated radiotherapy in locally advanced head and neck cancer. A phase I dose escalation study. AB - Both docetaxel and irinotecan have shown strong radiosensitizing properties in vitro. Encouraging results have been reported by phase I/II studies on combined docetaxel or irinotecan with radiotherapy. In the present study we investigated the feasibility of double radiosensitization with weekly docetaxel and irinotecan in head and neck cancer. Twelve patients with locally advanced squamous cell head and neck cancer were recruited in a phase I/II dose escalation protocol. Radiotherapy was given as a standard fractionation regimen (2 Gy/day, 5 fractions/week) to a total dose of 66-70 Gy. Three Docetaxel/Irinotecan dose levels were examined thus, 20/25 mg/m2 (level 1), 20/40 mg/m2 (level 2) and, 25/55 mg/m2 (level 3). Severe asthenia was observed in 1/4 patients treated in the 2nd dose level and in all 4 patients treated in the 3rd. The onset of severe asthenia was associated with the onset of severe grade 3/4 mucositis during the 4th week of treatment. Radiation induced mucositis was accompanied by fungal infection in all 5 patients. The symptomatology persisted for 10-14 days. Mild grade 2 mucositis was observed in 7/8 patients treated at the 1st and 2nd dose level, which enforced treatment delay for 3-5 days. Neutrophil toxicity was minimal. There was only one patient treated at the 3rd dose level that presented with grade 2 neutropenia. Hemoglobin toxicity was also minimal, showing a median drop of 1.2 gr/dL. There was no platelet toxicity observed. Complete response was observed in 9/12 (75%) patients and partial response was observed in 3/12 patients. Of interest, the lowest CR rate was observed in the 3rd dose level (2/4; 50%), which may be a consequence of overall treatment time prolongation. It is concluded that docetaxel and irinotecan combination with radiotherapy is feasible and, a high CR rate can be expected. Combination of the regimen with cytoprotective agents warrant further investigation. PMID- 10472350 TI - Evaluation of chemotherapy response in patients with small cell lung cancer using Technetium-99m-tetrofosmin. AB - The aim of this study was to investigate the relationship between Technetium-99m (Tc-99m) tetrofosmin accumulation in tumors which may represent the expression of multidrug resistance (MDR)--mediated P-glycoprotein (Pgp) and multidrug resistance-associated protein (MRP) as well as response to chemotherapy in patients with untreated small cell lung cancers (SCLC). Twenty patients with SCLC were studied with Tc-99m tetrofosmin lung scintigraphy before chemotherapeutic induction. Tc-99m tetrofosmin lung scans were interpreted both visually and quantitatively. Response was evaluated upon completion of chemotherapy. Patients with good chemotherapy response had a significantly higher incidence (93%) of positive Tc-99m tetrofosmin lung single photon emission computed tomography (SPECT) findings than patients with poor response (33%) (p value < 0.05). The tumor/background ratios (T/B) were 1.8 +/- 0.4 and 1.2 +/- 0.3, for patients with good response and poor response, respectively (p < 0.05). However, other prognostic factors (tumor size and stage) were not significantly related to Tc 99m tetrofosmin lung scan findings and chemotherapy responses. Tc-99m tetrofosmin lung scintigraphy can predict chemotherapy response in patients with SCLC. PMID- 10472351 TI - Peritonectomy combined with intraperitoneal chemohyperthermia in abdominal cancer with peritoneal carcinomatosis: phase I-II study. AB - OBJECTIVE: To evaluate the feasibility and the tolerance of Peritonectomy Procedure (PP) combined with Intraperitoneal Chemohyperthermia (IPCH) in patients with peritoneal carcinomatosis, a phase I-II study has been realised from January 1997 to September 1998. METHODS: Eighteen patients were included for peritoneal carcinomatosis from colorectal cancer (13), ovarian cancer (2), gallbladder cancer (1), gastric cancer (1) and peritoneal mesothelioma (1). Peritoneal carcinomatosis were mainly advanced disease (16 stage 3 and 4, 2 stage 2). All the patients underwent surgical resection of their primary tumor with PP as described by Sugarbaker and IPCH (with Mitomycin C, Cisplatinum or both). IPCH used in this study was a "closed sterile circuit" device with inflow temperatures ranging from 46 to 48 degrees C. IPCH was performed on the same day as PP (8118) or delayed (10/18). RESULTS: Significant down-staging of peritoneal carcinomatosis was achieved for 16 patients. One patient died postoperatively, while the morbidity rate was 6/18 (long postoperative ileus, grade 3 leucopenia and anastomotic leakage). CONCLUSIONS: Combination of PP and IPCH could achieve significant tumoral volume reduction in peritoneal carcinomatosis. This aggressive treatment must be employed selectively because of its morbidity. Larger phase III studies are now needed. PMID- 10472352 TI - P53 antibody response in patients with vulvar cancer. AB - BACKGROUND: Mutations in the p53 tumor suppressor gene are the most common genetic changes identified in cancer cells. Several studies report alterations of the p53 gene in vulvar cancer. As observed in a wide variety of human malignomas, mutant p53 protein may provoke a specific humoral immune response. The possible occurrence of p53 antibodies in patients with vulvar cancer has not been investigated so far. MATERIALS AND METHODS: We used a specific p53 antibody ELISA to investigate serum samples of 41 patients with vulvar cancer taken prior to therapy and serum samples of 17 healthy controls. RESULTS: Of the 41 patients with vulvar cancer, 4 (10%) were found to be positive for serum p53 antibodies, contrary to the 17 healthy volunteers without p53 serum antibodies (chi-square test, p = 0.2). No significant correlations were found between p53 antibody status and tumor stage (p = 0.64), histological grade (p = 0.89), and patients' age (p = 0.87). We found no significant association between the p53 serum antibody status in vulvar cancer patients and disease-free (p = 0.67) and overall survival (p = 0.7). CONCLUSIONS: In summary, it is the first time that p53 antibodies have been detected in the sera of patients with vulvar cancer. However, p53 serum antibodies did not serve as prognostic markers in vulvar cancer. PMID- 10472353 TI - Recent advances in early prostatic cancer. AB - Early prostatic carcinoma is a slowly progressing, localized malignant tumor which has been recently discovered with increased frequency due to the use of improved diagnostic methods. The combination of digital rectal examination, serum PSA level and transrectal ultrasound is currently the best available diagnostic tool, although other putative diagnostic markers and techniques are being investigated. Core needle biopsy may follow if there is suspicion of malignancy and in doubtful cases the most useful antibody for the immunohistochemical diagnosis of early, low grade prostatic carcinoma is clone 34 beta E12. Cytogenetic techniques and molecular biological methods are increasingly being used for further investigating localized prostate carcinomas in order to identify early molecular targets and alterations, which may lead to progression. Chromosome abnormalities, cell to cell and cell to matrix interactions, changes in the status of steroid hormone receptors, oncogenes and tumor suppressor genes, as well as other, as yet unclear, events may be of importance in prostate carcinogenesis and the progression of early malignant tumors to aggressive phenotypes. A variety of putative prognostic markers, apart from serum PSA levels, histological grade and tumor volume, such as neuroendocrine differentiation, angiogenesis, cell proliferation labeling index and ploidy analysis may prove useful in evaluating tumor progression in early prostatic carcinomas. The final and most important goal of all investigations related to early prostate cancer is to contribute to the best therapeutic management of the individual patient. PMID- 10472355 TI - Radiotherapy on adult patients with soft tissue sarcoma with fast neutrons or photons. AB - Through adjuvant photon radiotherapy, it is possible to improve tumor control as well as avoid ultra radical surgery in patients with soft tissue sarcoma. Similar results have been obtained in cases of incomplete resected G1/G2 sarcoma after applying neutron radiotherapy. We compared the results of a group of patients with soft tissue sarcoma treated with those having received photon therapy (100 patients) or neutron therapy (61 patients). The median dose in the photon treated group was 60 Gy (range 45 to 65 Gy). The neutron therapy group received a median dose of 14.1 Gy (range 5.0 to 18.57 Gy). Patients treated with mixed-beam irradiation received an average dose of 36.5 Gy photon and 8.5 Gy neutrons. The 5 year survival rate of the photon group rated 43.1%. In the neutron group we found 42.5%, respectively. In both groups the results of surgical resection and grading were of high significance according to survival. 4% of the patients belonging to the photon group developed grade III/IV WHO side effects. In the neutron group side effects grade III/IV WHO were observed in 11% of the cases. Comparing treatment results of neutron and photon therapy we demonstrated that incompletely resected G1 and G2 tumor patients show greater benefit in the case of neutron radiotherapy. PMID- 10472354 TI - 99mTc-Sestamibi scanning with SDZ PSC 833 as a functional detection method for resistance modulation in patients with solid tumours. AB - BACKGROUND: Our aim was to determine the value of 99mTc-Sestamibi scanning as functional detection method of P-glycoprotein (Pgp) blockade by PSC 833 in solid tumour patients. PATIENTS AND METHODS: Day 1 and day 4 after 2,200 mg orally administered PSC 833 the tumour area was scanned after intravenous (i.v.) administration of 400 MBq 99mTc-Sestamibi. In tumours with net 99mTc-Sestamibi uptake and in the hepatic region K-efflux was determined. Whole blood was analyzed for 99mTc-Sestamibi, and PSC 833 levels. RESULTS: Fourteen patients were included. In the only Pgp-positive tumour with positive 99mTc-Sestamibi scanning K-efflux of 99mTc-Sestamibi decreased significantly after PSC 833 intake. A net inhibition of liver efflux of Sestamibi after PSC 833 intake was observed in all evaluable patients. PSC 833 blood levels were all above 2 mg/L during scanning; 99mTc-Sestamibi blood levels post versus pre PSC 833 were unchanged. CONCLUSIONS: PSC 833 induced modulation of K-efflux of 99mTc-Sestamibi in a Pgp positive tumour and in all patients in the liver. PMID- 10472356 TI - Non-polypoid colorectal neoplasias: a multicentric study. AB - A total of 781 non-polypoid colorectal neoplasias harvested at 4 main Hospitals in Tokyo, Japan (n = 420) and at 4 different time-intervals at the Karolinska Hospital, Stockholm, Sweden (n = 361) were reviewed. By applying strict histologic definitions, the lesions were classified into adenomas with low grade dysplasia (LGD), with high grade dysplasia (HGD), intramucosal carcinomas (IMC) or submucosal carcinomas (SMC). Of the non-polypoid neoplastic lesions reviewed in Sweden, 82.8% (n = 299) had LGD. In Japanese patients only 42.6% (n = 179) had LGD (p < or = 0.001). On the other hand, as many as 42.4% (n = 178) of the non polypoid lesions in Japanese patients had HGD, but only 14.1% (n = 51) of those in Swedish patients (p < or = 0.001). Whereas 15.0% (n = 63) of the non-polypoid neoplasias seen in Japan were IMC or SMC, only 3.0% (n = 11) of those seen in Sweden were IMC or SMC (p < or = 0.001). The cause(s) for these differences remains unclear. In Japan, however, a marked increased incidence of colonic cancer has been recorded in later years. Whether the "catching up phenomenon" by the Japanese with western colonic cancer incidence includes increased histologic aggressiveness of non-polypoid neoplastic polyps--as found in this survey- remains to be elucidated. PMID- 10472357 TI - Immunohistochemical detection of matrix metalloproteinases (MMP) 1 and 2, and tissue inhibitor of metalloproteinase 2 (TIMP 2) in stage I and II endometrial cancer. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are a family of zinc-dependent metalloendopeptidases which participate in the degradation of collagen and other extracellular matrix macromolecules. Expression of gelatonic MMPs, such as MMP-2 has been linked to enhanced tumor invasion and metastasis in in vitro and in vivo model systems. It was the aim of this study to determine whether the expression of MMP-1, MMP-2, and TIMP-2 correlates with survival in patients with surgically treated endometrial cancer. METHOD: A sample of 103 paraffin-embedded tumor specimens of surgical treated endometrial cancer was immunohistochemically investigated. RESULTS: MMP-1, MMP-2, and TIMP-2 were detected by immunohistochemistry in 95% (98/103), 87% (89/103), and 80% (82/103) of the tumour samples, respectively. Correlation coefficients for MMP-1/MMP-2, MMP 1/TIMP-2, MMP-2/TIMP-2 were 0.28 (p = 0.004), 0.05 (p = 0.6), and -0.03 (p = 0.73), respectively. In the univariate analysis, the expression of MMP-1 (log rank test, p = ns), MMP-2 (log-rank test, p = ns), and TIMP-2 (log-rank test, p = ns) were not associated with overall survival. CONCLUSION: MMP-1, MMP-2, and TIMP 2, detected by immunohistochemistry are not helpful in predicting the prognosis of endometrial cancer patients. PMID- 10472358 TI - A profile of tumor markers in the population of Kuwait. AB - The tumour markers CEA, AFP, CA 125 and CA 199 were analyzed in a group of apparently healthy subjects in Kuwait. The sample (n = 394) included both genders in the population with a mean age of 38 (S.d. 12.0) years. The distribution of CEA levels values was significant different (Mann-Whitney U test) between Kuwaiti and non-Kuwaiti. The distribution of AFP levels was found to be the same in all groups. The distribution of CA 125 levels was significantly higher in females than in males, both in Kuwaiti and non-Kuwaiti. The distribution of CA 19-9 values was found to be significantly higher in the Kuwaiti female group when compared to the males. The upper reference level was defined as the 95 percentile of the normal values in each group. In the total population the reference level of AFP was 5.6 micrograms/l and of CA 19.9 43 kU/l. The reference level of CA 125 was 16 U/l in males and 24 kU/l in females, respectively. The CEA reference level in Kuwaitis was 6.9 micrograms/l and in non-Kuwaitis 4.4 micrograms/l. The results indicated the importance of determining the reference levels of tumour markers for each individual laboratory. It was also emphasized that care should be taken on the impact of 95 percentiles of normal and benign disease groups. PMID- 10472359 TI - Poor outcome of patients with resectable breast cancer receiving adjuvant high dose sequential chemotherapy following preoperative treatment. AB - BACKGROUND/OBJECTIVES: The prognosis of resectable high risk breast cancer (BC) patients (N+ > 10) is poor with a five-year disease-free survival (DFS) after standard adjuvant ADM/CMF chemotherapy (CT) of about 40%. An improvement in survival has been reported when high-dose chemotherapy with autologous stem cell support is given. It has been recently suggested that nodal status and the degree of pathological remission following preoperative CT administered in patients harbouring tumors larger than 3 cm represent the most important prognostic factors for DFS. Since no data are available regarding the impact of primary CT in the high dose CT adjuvant setting, we retrospectively evaluated the efficacy of administering megadoses of cytotoxic drugs with stem cell support in the subgroup of patients showing poor response to preoperative CT., PATIENTS AND METHODS: Fourteen women with high risk BC, N+ > 10 and tumor size > 3 cm following antracyclin-based primary CT, received high dose sequential chemotherapy (HDS). The median number of positive axillary nodes at surgery was 18 and tumor size was greater than 5 cm in 6 patients. HDS chemotherapy consisted of cyclophosphamide (7 gr/m2), methotrexate (8 gr/m2) plus vincristin (2 mg), 2 courses of carboplatin (360 mg/m2), and Thiotepa (600 mg/m2) plus L-PAM (160 mg/m2) as final myeloablative regimen requiring stem cell support. RESULTS: At a minimum follow up of 12 months (median 18 months, range 12-40) 5 patients remained disease free (36%) and 9 (64%) have relapsed (7 within the first 10 months). CONCLUSION: Our retrospective analysis suggests that BC patients showing poor response to primary CT might fail to achieve the benefits expected from high dose intensification. PMID- 10472361 TI - Gastric adenocarcinomas in displaced mucosal glands. AB - Although gastric carcinoma is one of the most common cancers worldwide, only a few histologic proximate precursors have been demonstrated. Several authors have found that foci of misplaced gastric mucosa with cystic dilatations (s.c. heterotopic mucosa) are often associated with gastric adenocarcinomas. However, adenocarcinomas originating within heterotopic gastric mucosa have never been reported. In present work, the review of 213 consecutive gastrectomy specimens in Japanese patients showed heterotopic gastric mucosa in 20.1% (n = 43). Up to 18 foci per gastrectomy were present. The heterotopic mucosa was surrounded by invaginations of the muscularis mucosae which showed strong positivity for smooth muscle actin. In 3 of the 213 specimens, an adenocarcinoma was found within a focus of heterotopic gastric mucosa. All 3 adenocarcinomas had cystic dilatations lined by neoplastic columnar epithelium with polymorphic nuclei, irregular nuclear membrane, large irregular nucleoli and pathological mitosis. The tumors had lateral bundles of smooth muscle (smooth muscle actin positive), regarded as invaginations of the muscularis mucosae. The p53 protein was strongly overexpressed in all 3 tumors. The heterotopic gastric mucosa may be one mucosal locus from which gastric adenocarcinomas may originate. PMID- 10472360 TI - Frequency of p53 codon 72 genotypes in human papillomavirus associated squamous intraepithelial lesions and cervical cancer. AB - BACKGROUND: A recent study suggested that the p53Arg (at residue 72) homozygous genotype could be a potential genetic risk factor for cervical cancer among white women. To confirm this result we examined the proportion of p53 genotypes in a larger number of patients with cervical cancer and in patients with squamous intraepithelial lesions (SIL) compared to a control population. MATERIALS AND METHODS: We used allele-specific primers to amplify the p53Arg and p53Pro sequences and we examined the proportion of genotypes in the study populations using chi 2-test. RESULTS: The distributions of p53Arg homozygous, heterozygous and p53Pro homozygous genotypes were 63%, 27% and 10% in cervical cancer patients, 53%, 36% and 8% in individuals with SIL, and 60%, 36% and 4% in control population. Using chi-square test, no significant difference was found between genotype frequencies in the study groups. CONCLUSION: Thus, the p53Arg homozygous genotype does not seem to increase the risk of cervical cancer in Hungarian women. PMID- 10472363 TI - Patient contact is the major determinant in incident leprosy: implications for future control. AB - Notwithstanding the elimination efforts, leprosy control programs face the problem of many leprosy patients remaining undetected. Leprosy control focuses on early diagnosis through screening of household contacts, although this high-risk group generates only a small proportion of all incident cases. For the remaining incident cases, leprosy control programs have to rely on self-reporting of patients. We explored the extent to which other contact groups contribute to incident leprosy. We examined retrospectively incident leprosy over 25 years in a high-endemic village of 2283 inhabitants in Sulawesi, Indonesia, by systematically reviewing data obtained from the local program and actively gathering data through interviews and a house-to-house survey. We investigated the contact status in the past of every incident case. In addition to household contact, we distinguished neighbor and social contacts. Of the 101 incident cases over a 25-year period, 79 (78%) could be associated to contact with another leprosy patient. Twenty-eight (28%) of these 101 cases were identified as household contacts, 36 (36%) as neighbors, and the remaining 15 (15%) as social contacts. Three patients had not had a traceable previous contact with another leprosy patient, and no information could be gathered from 19 patients. The median span of time from the registration of the primary case to that of the secondary case was 3 years; 95% of the secondary cases were detected within 6 years after the primary case. The estimated risk for leprosy was about nine times higher in households of patients and four times higher in direct neighboring houses of patients compared to households that had had no such contact with patients. The highest risk of leprosy was associated with households of multibacillary patients. The risk of leprosy for households of paucibacillary patients was similar to the risk of leprosy for direct neighboring houses of multibacillary patients, indicating that both the type of leprosy of the primary case and the distance to the primary case are important contributing factors for the risk of leprosy. Contact with a leprosy patient is the major determinant in incident leprosy; the type of contact is not limited to household relationships but also includes neighbor and social relationships. This finding can be translated into a valuable and sustainable tool for leprosy control programs and elimination campaigns by focusing case detection and health promotion activities not only on household contacts but also on at least the neighbors of leprosy cases. PMID- 10472362 TI - p53 as a prognostic factor for stage III serous adenocarcinoma of the ovary. AB - Expression of mutant p53 in 19 patients with stage III moderately differentiated serous cystadenocarcinoma of the ovary, who all had optimal primary cytoreductive surgery and six cycles of cisplatin and cyclophosphamide was analyzed to determine its prognostic significance. All patients were followed up for a minimum of 48 months. Immunohistochemical analysis for mutant p53 was by means of monoclonal antibody DO7 and Pab240. Mean survival for patients expressing mutant p53 was 44 months and 117 months for those not expressing mutant p53. In our study, expression of mutant p53 proved to be a statistically adverse prognostic factor (p = 0.029). PMID- 10472364 TI - Reanimation of the lagophthalmos using stainless steel weight implantation; a new approach and prospective evaluation. AB - Weight implants have been used for over 30 years in surgical rehabilitation following lagophthalmos. However, there has been a renewed interest among ophthalmologists in recent years. The weight implant technique combined with lateral canthus resuspension was performed on 22 eyes in Thai patients. It is a reliable method for correcting lagophthalmos and ectropion. In previous studies almost pure gold was used in 99.99% of eyelid implants. However, we wanted to use other materials which are not expensive and are readily available. A stainless steel plate was selected. The majority of our surgeries were done with a stainless steel weight implant of 1.368 g. Patients were followed for 7 to 55 months, and no rejections of the implants were observed. This study achieved a > or = 90% success rate, and showed that a stainless steel plate is an alternative to gold for the weight implant technique. PMID- 10472365 TI - Lymphoproliferative responses of leprosy patients and healthy controls to nitrocellulose-bound M. leprae antigens. AB - The lymphoproliferative responses of 51 leprosy patients and 11 healthy contacts were analyzed using the nitrocellulose-bound specific antigen fractions from the cell-free extract of Mycobacterium leprae. The main proliferation-inducing fraction for peripheral blood mononuclear cells of the healthy contacts was found to be the Fraction II, bearing antigens in the range of 66-45 kDa. However, this fraction failed to induce lymphoproliferation in the leprosy patients, unlike healthy contacts (p < 0.032). The number of responders as well as the strength of the responses to 66-45 kDa proteins were found to be low in the leprosy patients compared to the healthy contacts. Further, preliminary analysis with the subfractions of Fraction II produced a similar pattern, suggesting that the immune response to the antigens in the range of 66-45 kDa M. leprae proteins remains suppressed in subjects with clinical signs and symptoms of the disease. PMID- 10472366 TI - Lot quality assurance sampling (LQAS) for monitoring a leprosy elimination program. AB - In a statistical sense, prevalences of leprosy in different geographical areas can be called very low or rare. Conventional survey methods to monitor leprosy control programs, therefore, need large sample sizes, are expensive, and are time consuming. Further, with the lowering of prevalence to the near-desired target level, 1 case per 10,000 population at national or subnational levels, the program administrator's concern will be shifted to smaller areas, e.g., districts, for assessment and, if needed, for necessary interventions. In this paper, Lot Quality Assurance Sampling (LQAS), a quality control tool in industry, is proposed to identify districts/regions having a prevalence of leprosy at or above a certain target level, e.g., 1 in 10,000. This technique can also be considered for identifying districts/regions at or below the target level of 1 per 10,000, i.e., areas where the elimination level is attained. For simulating various situations and strategies, a hypothetical computerized population of 10 million persons was created. This population mimics the actual population in terms of the empirical information on rural/urban distributions and the distribution of households by size for the state of Tamil Nadu, India. Various levels with respect to leprosy prevalence are created using this population. The distribution of the number of cases in the population was expected to follow the Poisson process, and this was also confirmed by examination. Sample sizes and corresponding critical values were computed using Poisson approximation. Initially, villages/towns are selected from the population and from each selected village/town households are selected using systematic sampling. Households instead of individuals are used as sampling units. This sampling procedure was simulated 1000 times in the computer from the base population. The results in four different prevalence situations meet the required limits of Type I error of 5% and 90% Power. It is concluded that after validation under field conditions, this method can be considered for a rapid assessment of the leprosy situation. PMID- 10472367 TI - Review of leprosy control activities in Yemen. AB - Leprosy control activities in Yemen are reviewed historically and up to the present time. Since 1983 the World Health Organization's multidrug therapy has been used in the National Leprosy Control Programme. Current activities are carried out in 63 leprosy clinics distributed all over the country and staffed by trained primary health care workers and medical assistants. In Yemen leprosy prevalence has declined from 1400 per 10,000 population in 1990 to 647 in 1997. Over the same period, new case detection rates per 10,000 population increased from 185 to 517. A backlog of leprosy cases continues to transmit the disease in Yemen. PMID- 10472368 TI - Effect of oil of hydnocarpus on wound healing. AB - Oil of hydnocarpus has been replaced by other chemotherapeutic agents which have a better mycobactericidal effect. However, none of the currently used antileprosy drugs has been reported to have a positive effect in wound healing. Anecdotal reports claim that leprosy patients who have taken capsules containing oil of hydnocarpus orally have shown more rapid wound healing than those not receiving it. In view of these reports, a pilot experimental study was undertaken to determine the effect of the oil of hydnocarpus in wounds experimentally inflicted on male Wistar rats. The wound-healing effect of oil of hydnocarpus was studied with reference to collagenation and the strength of the scar tissue. The drug treated group showed a significant increase in body weight and strength of scar tissue in the incision model and, also, increased strength of the collagen tissue and hydroxyproline content in the dead space model. The results of this pilot study indicate that the oil of hydnocarpus, which also has antileprotic activity, could be a useful adjunct in the healing of wounds and ulcers in leprosy patients. PMID- 10472369 TI - Efficacy of WHO regimens in the management of leprosy patients with G6PD deficiency. PMID- 10472370 TI - Infection by M. leprae is governed by the temperature at the entry point: a preliminary note. PMID- 10472371 TI - "Large numbers" of Mycobacterium leprae are discharged from the intact skin of lepromatous patients; a preliminary report. PMID- 10472372 TI - Staging nerve involvement in M. leprae infection. PMID- 10472373 TI - Detection of antibodies toward secreted mycobacterial antigen 85 in untreated leprosy patients' sera. PMID- 10472374 TI - Dapsone syndrome: hepatitis-B infection a risk factor for its development? PMID- 10472375 TI - Granuloma multiforme (called Nkanu disease in the 1940s and Mkar disease in 1964) PMID- 10472376 TI - Histoid leprosy; report of three cases. PMID- 10472377 TI - HMGI(Y) and HMGI-C dysregulation: a common occurrence in human tumors. AB - HMGI(Y) and HMGI-C are members of a distinct family of "high mobility group" (HMG) proteins that are nonhistone chromatin-associated proteins initially characterized by high electrophoretic mobility in polyacrylamide gels (hence the acronym HMG). Recent studies have shown that HMGI(Y) and HMGI-C are important elements with a role in the regulation of chromatin structure and function. Like other HMG proteins they are responsible for the correct three-dimensional configuration of protein-DNA complexes and therefore play a key role in important cellular processes such as DNA transcription. Aberrant HMGI(Y) and HMGI-C expression generally correlates with a malignant tumor phenotype. However, HMGI(Y) and HMGI-C dysregulation, as a result of specific chromosomal rearrangements, is also being identified in a variety of common benign mesenchymal tumors such as lipomas and uterine leiomyomas making HMGI(Y) and HMGI C genes probably the most commonly rearranged genes in human neoplasms. While a precise definition of the HMGI(Y) and HMGI-C role in tumor initiation and progression is still missing, it is likely that future investigations will contribute valuable insights to the understanding of human neoplasia. PMID- 10472378 TI - Ovarian tumors of borderline malignancy (tumors of low malignant potential): a critical appraisal. AB - Before the designations borderline malignancy and low malignant potential were used, the surface epithelial-stromal tumors of the ovary were simply classified into benign and malignant categories. The introduction of the borderline category of tumors has been a great advancement in classification, because it has set apart from the general group of surface epithelial cancers a subgroup with a much better prognosis, stage-for-stage, than that of conventional ovarian carcinomas. Over the last 20 years, pathologists have learned to recognize the distinctive clinicopathologic features of serous borderline tumors as well as the adverse prognostic significance of the associated invasive peritoneal lesions, whether they may represent true implants or independent primary tumors. We have urged our surgical colleagues to search for the peritoneal lesions, and sample them meticulously, and advised our fellow oncologists not to administer adjuvant therapy to patients with noninvasive implants lacking malignant epithelial cells. There is now convincing evidence in the literature that the only fatal cases of serous borderline tumors are those associated with invasive implants, and chemotherapy is indicated only for these rare tumors. It has also been demonstrated that Stage I intestinal mucinous borderline tumors and noninvasive well-differentiated mucinous carcinomas both have an almost equally good prognosis. The current treatment for pseudomyxoma peritonei is still unsatisfactory, but we have recently learned that most of the mucinous ovarian tumors associated with pseudomyxoma peritonei are secondary to similar tumors of the appendix. In the remaining cases, however, the ovarian tumor appears to be responsible for the pseudomyxoma peritonei. Borderline tumors also exist in the endometrioid, clear cell, and Brenner surface epithelial categories, but these tumors have been too rare for clear delineation of their clinical and pathologic features. Recently, some investigators have proposed to abandon the borderline category and to return to the old benign-malignant classification system by dividing unevenly the borderline tumors into a larger group of atypical proliferative epithelial cystadenomas and a smaller category of recently described but still not well-characterized noninvasive carcinomas. In the author's opinion, such a recommendation is misleading because it ignores the possibility of rare but significant behavioral exceptions on each of these two groups of tumors. Furthermore, careful tumor staging is mandatory in both instances regardless of the type of terminology used. It is hoped that by keeping the borderline designation, knowledge on this group of ovarian tumors will continue to expand as it has been until now. PMID- 10472379 TI - Collagenous fibroma (desmoplastic fibroblastoma): a distinctive fibroblastic soft tissue tumor. AB - Collagenous fibroma is a distinct, benign, fibroblastic/myofibroblastic proliferation, probably neoplasm. It is a slow-growing tumor arising in predominantly subcutaneous tissue. This tumor is composed of stellate-shaped fibroblasts and abundant interstitial collagen. Since none of reported collagenous fibromas recurred, simple excision is an appropriate treatment. Collagenous fibroma should be differentiated from fibromatosis, which has a high risk of local recurrence if simple local excision is done. Fibromatosis is more cellular and shows short fascicular arrangements of tumor cells and greater infiltration at the periphery than collagenous fibroma. PMID- 10472380 TI - Another dissertation on poorly differentiated carcinomas: is it really necessary? AB - Descriptions of poorly differentiated thyroid carcinomas, tumors with outcomes midway between that associated with well-differentiated and anaplastic carcinomas, have been prominent in the literature of the recent past. The neoplasms are characterized by the finding of poorly differentiated areas within papillary and follicular carcinomas. The work of Sakamoto clearly separates a group of poorly differentiated carcinomas, based on outcomes, from well differentiated and anaplastic forms by the identification of schirrous, solid, and trabecular areas within papillary and follicular carcinomas. However, the presence of similar areas within more "benign" forms of papillary and follicular carcinomas may obviate the easy identification of such neoplasms in daily practice. This article presents a refinement of Sakamoto's classification and although suffering from the same limitations, may be of some practical value. PMID- 10472381 TI - Sialoblastoma and epithelial tumors in children: their morphologic spectrum and distribution by age. AB - This commentary addresses the histologic spectrum of salivary gland neoplasms in children with emphasis on perinatal tumors. Histopathologically, perinatal tumors fall into four categories: 1) histologically benign with adult counterpart, 2) hamartomas 3) embryomas-sialoblastomas, and 4) histologically and biologically malignant adult analogue tumors. Although the criteria to serrate benign from malignant sialoblastomas are not well-established, the following histologic features would favor an aggressive clinical course: perineural and/or vascular spaces invasion, necrosis, and a cytologic atypia beyond the expected for embryonic epithelium. PMID- 10472382 TI - Skills, behaviors, and motivation serve as a framework for competency. PMID- 10472383 TI - Inadvertent hypothermia in the OR. AB - Hypothermia is one of the most common problems that patients experience in the perioperative environment. Hypothermia causes more problems for the patient than the mere unpleasant experience of feeling cold. Additional nursing interventions are required for a patient suffering from inadvertent hypothermia, which often delays the patient's discharge from the perioperative setting. Many of the nursing interventions that are successful in preventing hypothermia and restoring body heat to surgical patients, such as applying a warmed blanket to a patient's body, seem straightforward. Understanding the cause and effects of inadvertent hypothermia, however, is essential in recognizing it and implementing the most effective nursing interventions. PMID- 10472384 TI - Mad cow disease--the OR connection. AB - Creutzfeldt-Jakob disease (CJD) is one of the transmissible spongiform encephalopathies, a group of fatal, neurodegenerative disorders affecting both humans and animals. The causative agent is the prion, which is still being researched and is controversial. In the 1980s, bovine spongiform encephalopathy brought much media attention to these diseases. Bovine spongiform encephalopathy is the result of faulty industrial practices that produced cattle feed contaminated by prions. In the 1990s, a new variant of CJD (i.e., nvCJD) appeared in Britain. Researchers believe that nvCJD was passed to humans through oral consumption of contaminated beef. This article describes the history, causative agent, mode of transmission, clinical features and course, diagnosis, treatment, and decontamination and sterilization guidelines for this baffling disease. PMID- 10472385 TI - Regulatory statutes and issues--clinical accountability in perioperative settings. AB - Nursing regulations exist to protect and benefit the health, safety, and welfare of the individual. Accountability is a multifaceted concept with many interpretations. All nurses are expected to engage in professional role activities appropriate to their education, experience, and position. Ultimately, nurses are accountable to themselves, their patients, and their peers for their professional nursing actions. This article addresses regulatory statutes and other issues of importance in the perioperative setting, including ethical and legal considerations, patient advocacy, delegation of tasks to unlicensed assistive personnel, and selected expanded roles in perioperative nursing practice. PMID- 10472386 TI - Moveable intraoperative magnetic resonance imaging systems in the OR. AB - Recently, the Seaman Family Magnetic Resonance Research Center, Calgary, Alberta, Canada, unveiled leading technology with its moveable intraoperative magnetic resonance imaging (MRI) system. The new system, which is housed in the OR, allows surgeons to use well-established neurosurgical techniques and instrumentation with the convenience of moving the high-resolution magnet in and out of the surgical field at any time. This mobility provides surgical team members with updated images that are vital to ensuring quality and determining the effect of surgery on brain structure and function. Based on experiences with the first 48 patients, this article describes the intraoperative MRI method and provides pertinent guidelines for safe perioperative care using this innovative MRI system. PMID- 10472387 TI - Automating the supply chain in the OR. AB - At the University of Louisville (Ky) Hospital, staff members from the materials management and surgery departments have worked together to automate the supply chain. The goals were to remove supply activities from clinical staff members whenever possible, obtain and apply information for better product ordering and use, reduce personnel in the materials management department, and improve perioperative nurses' ability to obtain supplies--all at a decreased cost to the facility. This article describes how, after implementing point-of-service technology for all surgical supplies, the facility realized a cost-per-procedure savings of 16% and increased satisfaction among staff members in both departments. PMID- 10472388 TI - Emotional quotient management as a dynamic approach to challenges. AB - As patient advocates, perioperative nurses must view all patients as individuals and meet every action and response with a commitment to providing individualized quality care. To do this, it is essential for perioperative nurses to overcome unproductive thought processes that can negatively affect their daily practice and how they view their jobs. This article explores an approach called Emotional Quotient Management, which uses six steps to teach people to overcome negative emotions, stay in control, and remain focused on success and happiness. PMID- 10472389 TI - Recommended practices for sterilization in perioperative practice settings. Association of periOperative Registered Nurses. PMID- 10472390 TI - OR gowns--even a "pass" can fail. PMID- 10472391 TI - Policy analysis as a strategy for clinical decision making. PMID- 10472392 TI - [Resilience: a new look at medical care and prevention]. PMID- 10472393 TI - [Epidemiologic study of multihandicapped children in the north of France]. AB - PATIENTS AND METHODS: The files of 440 severely handicapped children in the north of France were studied. These children had severe mental retardation and very poor motricity development. The methods of care and nursing were studied. RESULTS: Amounts these patients, 40.4% were in medico-social centers, 30% in non medico-social centers, and 29.4% lived at home with their parents. Only 6% were unknown by the departmental commission of specialized education (CDES). The rate of prevalence of the severely handicapped children from 5 to 19 years in the north of France was 0.7 per thousand. CONCLUSIONS: Results are compared with those from an INSERM study about handicap patients in 14 French counties. PMID- 10472394 TI - [Allergy to protein hydrolysates. Report of 30 cases]. AB - AIM OF THE STUDY: To describe the population of children allergic to cow's milk protein and their usual substitutes made of protein hydrolysates, and who are efficiently taken care of by using Neocate, an amino acid based formula. PATIENTS AND METHODS: Allergy to protein hydrolysates was diagnosed in 30 infants, aged from 15 d to 13 months (median = 3 months), who remained symptomatic while receiving a protein hydrolysate for 15 d to 12 months (median = 1.8 months). After a complete clinical and biological evaluation, an amino acid based diet using Neocate was attempted, followed after one month by an oral challenge test with the protein hydrolysate. RESULTS: Symptoms occurred mainly in the digestive tract (25 cases) in the form of regurgitations (18 cases), diarrhea (17 cases) and colicky pain (10 cases); failure to thrive was seen in ten cases. Neocate improved the clinical condition rapidly (within 3-10 d), and allowed the children to gain weight (27.5 +/- 10.8 g/d). Skin prick tests with cow's milk hydrolysates were positive in 13 out of 26 children; total IgE was raised in cases of cutaneous symptoms (six cases), and associated with increased specific IgE for cow's milk. During the intestinal permeability test, the lactitol/mannitol ratio which initially increased, decreased following Neocate (6.58 +/- 2.92% vs. 3.47 +/- 1.58%, P = 0.0004). Multiple food allergies were present in 22 cases. During challenge with a hydrolysate, the clinical reaction was immediate in 13 cases, partially delayed in 13 cases and delayed in four cases. CONCLUSION: Allergy to cow's milk hydrolysates may occur and has to be considered in the presence of anaphylaxis and also chronic digestive symptoms such as regurgitations, diarrhea and colicky pain, when these symptoms persist during cow's milk free diet. PMID- 10472395 TI - [Utilization of an equimolar mixture of oxygen-nitrous oxide in a general pediatric ward]. AB - We report our experience of the utilization of the 50% oxygen-nitrous oxide mixture (nitrous oxide 50%) in our general pediatric ward after one year of use. PATIENTS AND METHODS: Between 1st April 1997 and 31st March 1998, children who had to undergo a painful procedure were proposed to inhale 50% nitrous oxide before the procedure. We evaluate pain, restlessness and adverse effects. RESULTS: The procedures (127 of them) were carried out in 90 children (61 boys). They were aged from 5 months to 15 years (mean: 5.7 years; median: 4.1 years). Indications were: lumbar puncture (n = 45), burning dressing (n = 29), venous cannulation (n = 12), minor surgery (n = 27), and miscellaneous (n = 14). Inhalation time was between 2 to 70 min (mean: 14.4 min; median: 11 min). Pain was absent or low in 106 cases (83.4%). Restlessness was absent or low in 100 cases (78.8%). Averse events were observed 12 times, but they were always minor and quickly reversible. CONCLUSION: Nitrous oxide (50%) can be used successfully in a general pediatric ward. Other studies are necessary to define the best conditions. PMID- 10472396 TI - [Prevention of doping in sport in adolescents: evaluation of a health education based intervention]. AB - BACKGROUND: The extent of sport doping and its potential risks for health is to be considered as a problem of public health. Until now, only a very few number of prevention programs have been shown to be efficient. The aim of this prospective study was to test the benefit of a specific educational intervention to reduce the intent of adolescent athletes to use drugs. PATIENTS AND METHODS: The participants were 476 adolescent athletes involved into high-level events, 238 were included in the experimental group and 238 as control group. The intervention was divided in four stages: information on epidemiology of doping in sport, discussions about nutritional supplement and other authorized products, a drug simulating role play, and information on self-medication. The program was evaluated by a self-administrated questionnaire before, just after the intervention, and three months later. RESULTS: A significant decrease in the intention to use doping drugs, and an increase in the perception of being able to refuse doping was found in the experimental group three months after the intervention. CONCLUSIONS: This work suggests that health education based interventions are effective in preventing drug use among adolescent athletes. PMID- 10472397 TI - [Voluntary poisoning with a rodenticide in an adolescent]. AB - Suicide attempts are frequent during adolescence. Intentional ingestion of rat poison is not well known in France. The complications of this are prolonged and may be serious. CASE REPORT: An adolescent, 15 years old, with clinical hemorrhagic syndrome, had coagulation deficiency. Rat poison had been found in serum. The young girl recognized later that the ingestion of these toxins was intentional. CONCLUSION: Suicide attempt with rat poison is exceptional, but we have to mention it when vitamin K-dependent factors failed without any other explication. PMID- 10472399 TI - [Pneumothorax revealing cystic adenomatoid malformation of the lung in a 13 year old child]. AB - CASE REPORT: A 13 year old boy had a seven day history of chest pain and dyspnea. His right hemithorax was immobile with abolished breathing sounds. Initial chest X-ray revealed a right tension pneumothorax. A chest tube was inserted and the right lung re-expanded. However, despite two intrapleural injections of tetracyclin, the pneumothorax reappeared. Lung CT scan showed an intraparechymal cyst in the posterior part of the right upper lobe. Lobectomy was performed and histological study confirmed the diagnosis of type I cystic adenomatoid malformation of the lung. Two months after surgery, clinical and radiological examinations were normal. CONCLUSION: Spontaneous pneumothorax, as the initial manifestation of cystic adenomatoid malformation of the lung, is rare (three cases reported in children beyond the neonatal period, and two in adults). CT scan features correlate well with the pathologic features. Because of the risk of recurrent pulmonary infections and malignancy change, removal of the cystic lesions is advisable. PMID- 10472398 TI - [Idiopathic juvenile osteoporosis: a new observation]. AB - Idiopathic juvenile osteoporosis is a rare form of bone demineralization which occurs during childhood. The mechanism of disease remains unknown. We report a new case which illustrates the main difficulties of diagnosis and treatment. CASE REPORT: A 7 year old girl was admitted because of painful disability of her lower limbs. Diagnosis was based on radiological signs, total bone density and bone histologic pattern. Plasma levels of calcium, phosphorus, alkaline phosphatases, 25-OH D3 and parathormone were within the normal range value. Other diseases associated with bone demineralization, such as enteric malabsorption, endocrine or tumoral diseases, were excluded. Recovery occurred after some months of treatment with calcium, vitamin D and rehabilitation, but we could not establish a clear causal relationship. CONCLUSION: The relative role of increased bone resorption or defective osteoblast function remain to be discussed. Recovery often occurs with or without treatment, but sequelae can lead to disability. PMID- 10472400 TI - [Prenatal diagnosis of chylothorax]. AB - Chylothorax is defined as the presence of lymph in the pleural space. Congenital chylothorax is one of the most frequent causes of fetal pleural effusion. It may be primary or secondary. Careful assessment of the etiology and of possible associated anomalies is required. Main complications are pulmonary hypoplasia, hydrops fetalis and the risk of premature delivery. Management is still a mater of controversy, the diagnosis of fetal pulmonary hypoplasia being difficult in utero. Factors such as gestational age, evolution of pleural effusion on two weeks, signs of seriousness (hydrops fetalis), and pulmonary expansion after pleural puncture may help the physician to choose between abstention, pleural tapping or long-term in utero drainage. Post natal treatment consists of pleural drainage and assisted ventilation in cases of respiratory distress, correction of metabolic and immune disorders and exclusive parenteral nutrition. Once chylothorax is resolved, formula feeding without long-chain triglycerides is allowed. If pleural effusion persists despite a well conducted treatment, albumin infusion and diuretics may be used before considering surgical treatment. PMID- 10472401 TI - [Protein requirements in children during states of stress. Committee on Nutrition of the French Society of Pediatrics]. AB - Acute and chronic stress conditions affecting critically ill children are characterized by severe protein breakdown and growth failure. This paper describes the disorders of protein metabolism, and gives recommendations for protein and energy intakes during stress conditions in children. PMID- 10472402 TI - [Radiologic case of the month]. PMID- 10472403 TI - [Early discharge from maternity: problems, exigencies and recommendations]. AB - Early discharge 72 hours after delivery remains the mother choice, although the final decision is taken by the midwife as well as the paediatrician of the unit, and never before the third day. A follow-up at home is systematically proposed during at least two days. Specially trained midwives of the maternity are in charge of this follow up. A one year evaluation of this practice shows that early discharge, after good clinical evaluation, with regular follow up as long as necessary, is not responsible for infants rehospitalisations as a result of undiagnosed pathology or maternal incompetence. PMID- 10472404 TI - [Infant cries?!, tears??...and whispers]. AB - Although infant screaming and crying can be a symptom of medical or surgical pathology, it is also a way for the baby to express himself towards his family circle. Once excluded a possible organic disease, infant crying must be interpreted by assessing the family interaction and the way parents listen to their child; that does not mean substituting oneself to the parent's capacity for analysing and replying to the child's needs. PMID- 10472406 TI - [Kawasaki disease revealed by uveitis]. PMID- 10472405 TI - [Recurrent otitis: what is the place of complementary examinations]. AB - Recurrent otitis media is defined by the occurrence of three episodes of acute otitis media in a year. Several risk factors are now well recognized, particularly the occurrence before six months of age of a first episode of otitis media, day care centers, and upper respiratory tract infections (rhinopharyngitis). In most cases biological or radiological investigations are of little interest. They are only indicated in case of persistent recurrent episodes, despite eviction of all risk factors and adenoidectomy, and mainly in case of association with bronchopulmonary pathology. PMID- 10472407 TI - [Severe constriction of the visual field associated with vigabatrin discovered by thorough examination of a 17-year old girl]. PMID- 10472408 TI - [Tracing infants born to HIV positive mothers]. PMID- 10472409 TI - [Vaccination and pathologic status of children in a rural area in Senegal (Khombole)]. PMID- 10472410 TI - [Accidental ingestion of sildenafil (Viagra) by a child]. PMID- 10472411 TI - [Practical manufacture of a sucrose solution]. PMID- 10472412 TI - Schizophrenia as a neuronal illness. PMID- 10472413 TI - Evidence for altered trisynaptic circuitry in schizophrenic hippocampus. AB - Recent postmortem studies have demonstrated subtle alterations in the hippocampal formation (HIPP) of patients with schizophrenia (SZ). These changes include a decreased density of nonpyramidal neurons (NPs), an increase of the GABAA, but not benzodiazepine receptors and a neuroleptic-dose-related increase of GAD65-IR terminals, particularly in sectors CA3 and CA2. High resolution studies of the GABAA receptor have further suggested that a decrease of disinhibitory GABAergic activity (i.e., GABA-to-GABA) in stratum pyramidale of CA3 may coexist with reduced inhibitory modulation (i.e., GABA-to-excitatory pyramidal neuron) in the stratum oriens of this same sector. These changes could potentially involve excitotoxic damage to interneurons in CA2; but, the precise time frame for the induction of such an injury during pre- versus postnatal life cannot as yet be inferred from the available data. These findings are consistent with reports of abnormal oscillatory rhythms and increased basal metabolic activity in the HIPP of patients with SZ. The fact that patients with manic depression also show a decrease of NPs in CA2 suggests that changes in the GABA system may not be related to a susceptibility gene for SZ. Rather, these alterations could be associated with a nonspecific factor, such as stress, experienced either early in life or much later during adolescence or adulthood. Presumably, there are also changes associated in other transmitter systems that may play a more specific role in establishing the SZ phenotype. PMID- 10472414 TI - Neurochemical brain imaging investigations of schizophrenia. AB - Neurochemical brain imaging methods developed over the past 20 years offer significant promise for elucidating the biochemical underpinnings of schizophrenia. The two general methodologies used for these studies have been: 1) radiotracer imaging: PET (positron emission tomography) and SPECT (single photon emission computed tomography); and 2) NMR (nuclear magnetic resonance) imaging: fMRI (functional magnetic resonance imaging) and MRS (magnetic resonance spectroscopy). Despite conflicting findings, striatal D2 receptor density may be elevated in some, but not all patients. Elevated synthesis, and increased release of dopamine after amphetamine challenge have also been reported. Imaging of cortical 5-HT2A receptors suggests that this system is unaffected, in conflict with findings of postmortem studies. Although prior postmortem studies suggested an increase in cortical GABAA receptors, three SPECT studies have found no significant changes. MRS studies have shown decreased levels of NAA (N-acetyl aspartate) moieties in hippocampus and frontal cortex of schizophrenic patients, which is consistent with the reported loss of neurons and neuropil in postmortem brains. In conclusion, developments in radiotracer and NMR imaging have provided promising leads to the biochemical abnormalities associated with schizophrenia. Future significant understanding is likely to occur with the development of new probes and enhanced instrument technology, when applied with an appreciation of the heterogeneity of the disorder and the need for careful clinical assessment of patients. PMID- 10472415 TI - Altered GABA neurotransmission and prefrontal cortical dysfunction in schizophrenia. AB - Dysfunction of the dorsolateral prefrontal cortex appears to be a central feature of the pathophysiology of schizophrenia, and this dysfunction may be related to alterations in gamma aminobutyric acid (GABA) neurotransmission. Determining the causes and consequences of altered GABA neurotransmission in schizophrenia, and the relationship of these changes to other abnormalities in prefrontal cortical circuitry, requires an understanding of which of the multiple subpopulations of cortical GABA neurons are affected. The chandelier class of GABA neurons, especially those located in the middle layers of the prefrontal cortex (PFC), have been hypothesized to be preferentially involved in schizophrenia because they 1) receive direct synaptic input from dopamine axons, 2) exert powerful inhibitory control over the excitatory output of layer 3 pyramidal neurons, and 3) undergo substantial developmental changes during late adolescence, the typical age of onset of schizophrenia. Consistent with this hypothesis, the axon terminals of chandelier neurons, as revealed by immunoreactivity for the GABA membrane transporter, are reduced substantially in the middle layers of the PFC in schizophrenic subjects. This alteration appears to be selective for the chandelier class of GABA neurons and for the disease process of schizophrenia. These findings provide insight into the pathophysiologic mechanisms underlying prefrontal cortical dysfunction in schizophrenia, and they reveal new targets for therapeutic intervention in this illness. PMID- 10472416 TI - Structural brain imaging in schizophrenia: a selective review. AB - Structural neuroimaging studies have provided some of the most consistent evidence for brain abnormalities in schizophrenia. Since the initial computed tomography study by Johnstone and co-workers, which reported lateral ventricular enlargement in schizophrenia, advances in brain imaging technology have enabled further and more refined characterization of abnormal brain structure in schizophrenia in vivo. This selective review discusses the major issues and findings in structural neuroimaging studies of schizophrenia. Among these are evidence for generalized and regional brain volume abnormalities, the specificity of anatomic findings to schizophrenia and to men versus women with schizophrenia, the contribution of genetic influences, and the timing of neuroanatomic pathology in schizophrenia. The second section reviews new approaches for examining brain structure in schizophrenia and their applications to studies on the pathophysiology of schizophrenia. PMID- 10472417 TI - The physiological approach: functional architecture of working memory and disordered cognition in schizophrenia. AB - The method of single-cell recording in awake behaving monkeys as they perform behavioral tasks is perhaps the most powerful approach to understanding the neural basis of behavior. In contrast to cellular analyses in vitro, e.g., in slice preparations, the in vivo approach allows direct correlation of cellular activity and specific processes as they are isolated by behavioral paradigms. As brain mechanisms are studied under natural conditions, inferences about the dynamic basis of information processing are direct. Limitations of this approach include the inability to unequivocally identify the cell under investigation, though location by cytoarchitectonic area and cortical depth is possible. Neurophysiological studies have not only given a dynamic view of neural processing "on line" but have allowed investigators to examine fundamental issues about normal brain function and dementia. Here I illustrate some of these issues and the power of single cell physiology to address them. Before doing so, it should be patently obvious that recording one cell at a time is a technical necessity and in no way should be taken to imply that any behavior is dependent on a single cell. Rather the neuron investigated is a representative of a cohort of cells acting in aggregate. PMID- 10472418 TI - The nonlinear, complex sequential organization of behavior in schizophrenic patients: neurocognitive strategies and clinical correlations. AB - BACKGROUND: Thought disorder is a hallmark of schizophrenia and can be inferred from disorganized behavior. Measures of the sequential organization of behavior are important because they reflect the cognitive processes of the selection and sequencing of behavioral elements, which generate observable and analyzable behavioral patterns. In this context, sequences of choices generated by schizophrenic patients in a two-choice guessing task fluctuate significantly, which reflects an "oscillating dysregulation" between highly predictable and highly unpredictable subsequences within a single test session. In this study, we aimed to clarify the significance of dysregulation by seeing whether demographic, clinical, neuropsychological, and psychological measures predict the degree of dysregulation observed on this two-choice task. METHODS: Thirty schizophrenic patients repeatedly performed a LEFT or RIGHT key press that was followed by a stimulus, which occurred randomly on the left or right side of the computer screen. Thus, the stimulus location had nothing to do with the key press behavior. The range of key press sequence predictabilities as measured by the dynamical entropy was used to quantify the dysregulation of response sequences and reflects the range of fixity and randomness of the responses. A factor analysis was performed and step-wise multiple regression analyses were used to relate the factor scores to demographic, clinical, symptomatic, Wisconsin Card Sorting Test (WCST), and Rorschach variables. RESULTS: The LEFT/RIGHT key press sequences were determined by three factors: 1) the degree of win-stay/lose-shift strategy; 2) the degree of contextual influence on the current choice; and 3) the degree of dysregulation on the choice task. Demographic and clinical variables did not predict any of the three response patterns on the choice task. In contrast, the WCST and Rorschach test predicted performance on various factors of choice task response patterns. CONCLUSIONS: Schizophrenic patients employ several rules, i.e., "win-stay/lose-shift" and "decide according to the previous choice," that fluctuate significantly when generating sequences on this task, confirming that a basic behavioral dysregulation occurs in a single schizophrenic subject across a single test session. The organization or the "temporal architecture" of the behavioral sequences is not related to symptoms per se, but is related to deficits in executive functioning, problem solving, and perceptual organizational abilities. PMID- 10472419 TI - Pursuit tracking impairments in schizophrenia and mood disorders: step-ramp studies with unmedicated patients. AB - BACKGROUND: Eye tracking abnormalities are highly prevalent in schizophrenia, and are among the most promising phenotypic familial markers for the disorder. The neurophysiologic mechanisms underlying these disturbances and their diagnostic specificity for schizophrenia are not yet well characterized. METHODS: This study assessed eye tracking deficits using foveopetal and foveofugal step-ramp tasks (ramps moving toward and away from central fixation after an initial step respectively) across a range of target velocities in anti-psychotic-naive schizophrenia patients, previously treated but currently unmedicated chronic schizophrenia patients, unmedicated patients with either bipolar or unipolar mood disorders, and healthy volunteers. RESULTS: All patient groups demonstrated reduced pursuit gain in open loop and closed loop visual tracking conditions. There were no significant group differences in the latency or accuracy of catch up saccades on foveofugal ramp tasks. CONCLUSIONS: These findings indicate that open and closed loop pursuit eye movements are impaired during acute episodes of schizophrenia and mood disorders. The intact accuracy of saccades to moving targets in all patient groups indicates that an adequate representation of motion information is available to the saccade system. Therefore, pursuit disturbances in mood disorders and schizophrenia seem to result, at least in part, from a disturbance in sensorimotor integration in the pursuit system after the initial extraction of sensory motion information. No eye movement abnormalities observed during performance of step ramp tasks were specific to schizophrenia. PMID- 10472420 TI - Increased dopamine synthesis rate in medial prefrontal cortex and striatum in schizophrenia indicated by L-(beta-11C) DOPA and PET. AB - BACKGROUND: The aim of the present study was to investigate dopamine synthesis in the brain of drug-free schizophrenic patients, not only in the striatum but also in extrastriatal areas like the prefrontal cortex, brain areas that for a long time has been in focus of interest in the pathophysiology of schizophrenia. METHODS: PET was performed in 12 drug-free (10 drug-naive) psychotic schizophrenic patients and 10 healthy volunteers matched for age and gender using 11C-labelled L-DOPA as the tracer. The time-radioactivity curve from occipital cortex (located within Brodman area 17 and 18) was used as input function to calculate L-DOPA influx rate, Ki images, that were matched to a common brain atlas. A significant overall increase of the Ki values was found in the schizophrenic group as compared with healthy controls. RESULTS: In particular, significantly higher Ki were found in the schizophrenic patients compared to the controls in the caudate nucleus, putamen and in parts of medial prefrontal cortex (Brod 24). The Ki value reflect an increased utilization of L-DOPA, presumably due to increased activity of the amino acid decarboxylate enzyme. CONCLUSIONS: The results indicate that the synthesis of dopamine is elevated within the striatum and parts of medial prefrontal cortex in schizophrenia. PMID- 10472421 TI - Mitogen-activated protein kinases in schizophrenia. AB - BACKGROUND: Mitogen-activated protein kinases (MAPKs) are important mediators of signal transduction from the cell surface to the nucleus and have been implicated in the integration of a variety of physiologic processes in most cells, including neurons. To investigate the possible involvement of MAPKs in schizophrenia, we compared the levels of the MAPK intermediates in postmortem brain tissue obtained from schizophrenic and control subjects. Our focus was on the cerebellar vermis because of evidence suggesting that schizophrenia is associated with abnormalities of structure, function, and signal transduction in this brain region. METHODS: Cytosolic proteins were fractionated by gel electrophoresis and subjected to Western blot analysis using polyclonal MAPK antibody, which detects total extracellular signal-regulated kinases (ERKs) 1 and 2 levels, and monoclonal MAP kinase phosphatase (MKP) 2 antibody. RESULTS: Schizophrenic subjects had increased levels of ERK2 [2763 +/- (SD) 203 vs. 2286 +/- 607 arbitrary units, U = 17, p < .05] in cerebellar vermis. The levels of a dual specificity tyrosine phosphatase, MKP2, were significantly decreased in cerebellar vermis (1716 +/- 465 versus 2372 +/- 429 arbitrary units, U = 12, p < .02) from schizophrenic patients. ERK1/MKP2 and ERK2/MKP2 ratios in cerebellar vermis, but not in other brain regions, were significantly different in schizophrenic subjects as compared to control subjects (U = 15, p < or = .027; U = 3, p < .001, respectively). CONCLUSIONS: MAPK levels are elevated in the cerebellar vermis of schizophrenic subjects. This could result from a protein dephosphorylation defect in vivo and might be involved in the pathology of the disease. PMID- 10472422 TI - Hippocampal volume reduction in schizophrenia: effects of genetic risk and pregnancy and birth complications. AB - BACKGROUND: Hippocampal volume reduction has been repeatedly demonstrated in schizophrenia. The relative contribution of genetic and environmental factors to this is unclear. METHODS: To address this question, we compared volumetric measurements of the left and right hippocampus, obtained using stereological methods from brain MRI scans, from two groups of patients with schizophrenia as well as healthy controls (n = 26). Patients (n = 27) in the first group, had no family history of schizophrenia and had experienced severe pregnancy and birth complications (PBCs). The second group comprised of patients (n = 21) without a history of severe PBCs from families multiply affected with schizophrenia. RESULTS: Reduction of the left hippocampal volume was associated with the diagnosis of schizophrenia but was present only in patients with a history of severe PBCs; in this group the smaller hippocampal volume, the earlier the onset of psychosis. CONCLUSIONS: Our findings suggest that environmental factors, in this case severe PBCs, make a significant contribution to hippocampal abnormalities in schizophrenia. PMID- 10472423 TI - An MRI study of cerebellar vermis morphology in patients with schizophrenia: evidence in support of the cognitive dysmetria concept. AB - BACKGROUND: Cumulative evidence suggests the cerebellum is involved in cognition and may be important in the pathoetiology of schizophrenia. Functional imaging studies have identified a possible neural circuit that includes the cerebellum and may be abnormal in patients with schizophrenia, manifesting as a fundamental cognitive deficit conceptualized as cognitive dysmetria. To explore the role of the cerebellum in cognitive dysfunction and schizophrenia, this study was designed to evaluate the morphology of the cerebellar vermis, its relationship to other cortical areas, and to cognitive function in patients with schizophrenia. METHODS: Male patients with schizophrenia (n = 65) were matched by age and gender to 65 healthy male controls. Volume measures of the 4 cerebral lobes and total cerebellum were obtained using automated methods. The area of the cerebellar vermis (divided into three lobes) was traced on a midsaggital MRI slice. RESULTS: Patients had smaller frontal and temporal lobes. There were no group differences in total cerebellar volume. Patients had a smaller vermis area, accounted for by a smaller anterior lobe. The anterior vermis area was positively correlated with total cerebellar volume, temporal lobe volume, and FSIQ in patients, but not controls. CONCLUSIONS: These findings support the theory that regions of the cerebellum may be involved in a neural circuit that is structurally and functionally abnormal in patients with schizophrenia, leading to cognitive dysmetria. PMID- 10472424 TI - Caudate size in first-episode neuroleptic-naive schizophrenic patients measured using an artificial neural network. AB - BACKGROUND: Structural brain imaging studies have demonstrated an increase in caudate volume in schizophrenic patients medicated with typical neuroleptics and a volume decrease following treatment with atypical neuroleptics. The measurement of striatal volume in patients who have never been treated with neuroleptics may indicate whether these changes are superimposed on intrinsic basal ganglia pathology in schizophrenia or are solely neuroleptic-induced. METHODS: We studied 36 first-episode, neuroleptic-naive schizophrenic patients and 43 control subjects using an artificial neural network (ANN) to identify and measure the caudate nucleus. The resulting volumes were analyzed using an ANCOVA controlling for intracranial volume, age, gender, and socioeconomic status. RESULTS: The mean volume difference between the caudate nuclei of patients and control subjects was .297 mL, the caudate nuclei of the patients being smaller than those of controls. When we covaried for intracranial volume, this was a statistically significant difference in caudate volume (n = 79; df = 1,75; F = 4.18; p > .04). CONCLUSIONS: Caudate nuclei of neuroleptic naive schizophrenic patients are significantly smaller than those of controls. This suggests that patients suffering from schizophrenia may have intrinsic pathology of the caudate nucleus, in addition to the pathology observed as a consequence of chronic neuroleptic treatment. PMID- 10472425 TI - Genetic influence on auditory information processing in schizophrenia: P300 in monozygotic twins. AB - BACKGROUND: It is widely accepted that schizophrenia is to some extent genetically determined. Abnormalities of the P300 component are one of the most robust biological findings in schizophrenia. They outlast clinical impairment and are present also in relatives of schizophrenic patients. In the present study on schizophrenic twins, the heritability of auditory P300 abnormalities and the influence of task difficulty on heritability was examined. METHODS: Twenty-two monozygotic twin pairs were included into this study (eight pairs discordant, five pairs concordant for schizophrenia or schizoaffective psychosis according to ICD-10 criteria, and nine control pairs). Two different versions of the auditory oddball paradigm were used to control for deficient stimulus perception. RESULTS: Compared to healthy controls, P300 amplitudes were significantly smaller in affected twins as well as in the non-affected co-twins of the discordant pairs. CONCLUSIONS: Our results suggest that P300 amplitude reduction is a genetically transmitted vulnerability marker for schizophrenia. Because the findings were independent of the difficulty of the task and could be demonstrated even when pitch disparity was adjusted to the subjects' ability to discriminate tones, the findings can not be related to the genetic influence on higher information processing. PMID- 10472426 TI - Heterogeneity of anti-nuclear antibodies in autoimmune liver diseases. PMID- 10472427 TI - Parkinson's disease or Parkinson's diseases? PMID- 10472428 TI - The prevention of colorectal cancer by aspirin use. AB - Epidemiologic studies indicate strongly that aspirin use reduces the risk of colorectal cancer and adenoma by approximately 40 to 50%. Perhaps up to ten years of use may be required before a benefit is apparent in colorectal cancer. The chemo-preventive actions of aspirin and other non-steroidal anti-inflammatory agents (NSAIDs) in colorectal carcinogenesis are also supported by animal studies, and by intervention studies that demonstrate that the anti-inflammatory agent sulindac causes regression of adenomas in familial adenomatous polyposis. Despite this evidence, the clinical implications are not clear because of increased gastro-intestinal irritation and bleeding episodes related to chronic aspirin use. Emerging evidence suggests that the anti-tumor properties of NSAIDs may be related primarily to the inhibition of cyclooxygenase-2 (COX-2), one of the two isoenzymes of the COX enzyme family. If confirmed, a new generation of selective COX-2 inhibitors may retain some of the chemo-preventive properties of NSAIDs with fewer side-effects. Firm recommendations regarding the use of aspirin or other NSAIDs to prevent colorectal cancer must await further research. For now, the decision must lie with the patient, in consultation with his or her healthcare provider, after a careful weighing of all potential risks and benefits. PMID- 10472429 TI - Aspirin in the prevention of strokes. AB - Aspirin leads to a moderate but significant reduction of stroke, myocardial infarction (MI), and vascular deaths in patients with transient ischemic attack (TIA) and ischemic stroke. Low doses are as effective as high doses but are better tolerated in terms of gastro-intestinal side-effects. The recommended daily aspirin dose is therefore between 50 and 325 mg. Bleeding complications are not dose dependent and occur with the lowest doses. PMID- 10472430 TI - Current synthetic notions on aspirin induced asthma. AB - Aspirin was discovered by Gerhardt in 1853. The first examples of its negative side-effects in the form of asthma like dyspnea were described as early as 1911 by Gilbert, and then by Reeds and by Cookes. Widal depicted the typical symptomatic triad of aspirin-associated illness in 1922, with the symptoms 'nasal polyposis, severe asthma, and intolerance to aspirin'. The role of inhibition of the cyclooxygenase was highlighted in the physiopathologic interpretation of the diseases in 1975. Since then, great progress has been made not only in the analysis of the disease, but also in the way to treat it that has been performed thanks to a new approach in the relative positions of 'nose bronchi' that enables the development of a new strategy in the relation between 'nasal and chest specialists'. If the mean of immuno-allergologic investigations, and particularly the scanographic and endoscopic imaging, have enabled a better cure for 'illness due to aspirin', it is nevertheless the case that this illness remains cortico dependent too often. Important progress has been expected since the discovery of anti-leukotriens and several teams apparently emphasize how useful anti leukotriens are in the control of aspirin-associated illness. Thus, this enables one to hope for a decrease in the first effective dose of corticosteroids and ever for the total elimination thereof. PMID- 10472431 TI - Aspirin induced apoptosis in gastric cancer cells. AB - Aspirin has been well known for its anti-pyretic and anti-inflammatory action over the past century. Its main action in the gastro-intestinal tract has always been associated with erosion and ulceration. However in recent years, there has been evidence suggesting that aspirin and the more potent non-steroidal anti inflammatory drugs (NSAIDs), could reduce the risk of gastric and colon cancer. One of the possible mechanisms in chemo-prevention is the ability to induce apoptosis in epithelial cells of the gastro-intestinal origin. This article introduces the role of apoptosis in the body and the gastro-intestinal tract. Evidence on the chemo-preventive role of aspirin and NSAIDs are listed, and the mechanisms of action discussed. PMID- 10472432 TI - Receptor imaging with 111In-pentreotide and 123I-methoxybenzamide, and inhibition tests with octreotide and bromocriptine of mixed growth hormone/prolactin secreting pituitary tumors. AB - We have performed pituitary scintigraphy with 111In-pentreotide (OCT), a somatostatin analogue, and with metoxybenzamide (IBZM) by 123I-IBZM in two patients affected by mixed growth hormone/prolactin-secreting pituitary tumors. Short-term growth hormone (GH) inhibition by a single injection of OCT (100 micrograms s.c.), and short-term prolactin (PRL) inhibition by oral administration of 2.5 mg of bromocriptine (BCR), were also performed in both patients. The first patient, a 26 year old man, showed intense tumor uptake of 123I-IBZM scintigraphy, whereas 111In-OCT scintigraphy showed moderate tumor uptake. Five hours after the BCR inhibition test, a fall of 83% in PRL plasma levels (from 8,336 micrograms/L to 1,417 micrograms/L), and of 91.6% in GH plasma levels (from 39.5 micrograms/L to 3.3 micrograms/L) were observed. OCT inhibition test suppressed GH plasma levels from 36 micrograms/L to 3.5 micrograms/L. The patient was submitted to treatment with BCR and OCT. A dramatic shrinkage of the tumor was seen after six months of therapy. The lesion disappeared one year after the start of therapy. The second patient, a 64 year old man, showed intense uptake at 111In-OCT scintigraphy, while 123I-IBZM uptake was not observed. A test dose of BCR resulted in an acute fall of PRL (from 145 micrograms/L to 118 micrograms/L), but not of GH. A test dose of OCT decreased the GH plasma level from 61 micrograms/L to 4.5 micrograms/L. The patient was submitted to treatment with BCR and OCT that resulted in a computed tomography and magnetic resonance imaging decrease of 45% of tumor volume one year after the start of therapy. Our results suggest that both suppression tests with OCT and BCR, and scintigraphic studies in vivo with 123I-IBZM and 111In-OCT can be predictive for the effectiveness of therapies with dopamine agonists and/or SS-analogs in patients with mixed PRL/GH-secreting pituitary tumors. Further studies are required to evaluate the role of suppressive tests in selecting patients for appropriate clinical treatments. PMID- 10472433 TI - Induction of cellular immunosuppression by the human papillomavirus type 16 E7 oncogenic protein. AB - The human papillomavirus type 16 (HPV-16) E7 oncogenic protein is found in the culture supernatant of SiHa cells, a cervical carcinoma cell line. Extracellular E7 protein, acting as a viral toxin in human immune cells, induces the overproduction of the immune suppressive IFN alpha cytokine by APCs, and inhibits the T-cell response to recall and allogenic antigens. These effects should be taken into account for the design of anti-human cervical carcinoma vaccines. PMID- 10472434 TI - Clinical assessment of collagen cross-linked N-telopeptides as a marker of bone metabolism in patients with primary hyperparathyroidism. AB - This study assesses the clinical usefulness of collagen cross-linked N telopeptides (NTx), a new marker for bone resorption, as a marker for primary hyperparathyroidism. The study was conducted on 114 patients who had undergone parathyroidectomy for primary hyperparathyroidism. Urine NTx levels were longitudinally determined by ELISA. Longitudinal serum bone Gla protein (BGP) determinations and dual energy X-ray absorptiometry (DEXA) examinations were also performed before and after parathyroidectomy. Before parathyroidectomy, the mean urine NTx level in primary hyperparathyroidism was 149.0 +/- 116.8 pmol BCE/mumol Cr (normal: < 55 pmol BCE/mumol Cr), and the positive rate was 88.5%. The mean BGP level was 33.5 +/- 59.1 ng/mL (normal: 12.7 ng/mL), and the positive rate was 77.7%. The NTx positive rate was higher than the BGP positive rate. NTx values were significantly correlated with bone mineral density (P < 0.002) and BGP values (P < 0.005). Two weeks after parathyroidectomy the NTx levels had already decreased significantly to 65.1 pmol BCE/mumol Cr (P < 0.001), and by one month postoperatively they had fallen to 55.9 pmol BCE/mumol Cr, reaching the normal range much sooner than BGP. The NTx normalization rate two weeks postoperatively was 37%, and by 12 months postoperatively it was normal in 83% of patients. BGP levels decreased more gradually than the NTx levels, and only became normal in 9% of cases two weeks postoperatively, as opposed to 74% by 12 months. It is concluded that NTx reflects bone turnover in primary hyperthyroidism, and that it is an excellent marker for bony lesions before and after parathyroidectomy. PMID- 10472435 TI - Possible etiology of human breast cancer: comment. PMID- 10472436 TI - [Experimental transmission of Crimean-Congo hemorrhagic fever virus by Rhipicephalus evertsi evertsi (Acarina:Ixodidae)]. AB - We have conducted experiments to assess the ability of Rhipicephalus evertsi evertsi tick to transmit the Crimean-Congo haemorrhagic fever (CCHF) and determine their reproductive cycle. The Rh. e. evertsi was infected by intracoelomic (transparietal and intra-anal) inoculation during the imaginal stases and by oral feeding on an infected viremic goat during imaginal and nymphal stases. The infection rate, transovarial and trans-stasial CCHF virus transmission were monitored for virus reisolation after suckling mice inoculation and the virus identified by ELISA and IFA for antigen detection. After intracoelomic inoculation, unfed adults had viral titers ranging from 0.67 to 2.9 log DL50/0.02 ml and had transmitted the virus to their vertebrate hosts by blood feeding. After 8 to 10 days of blood feeding duration, infection rates were respectively 36% and 100% for male and female ticks. In two instances out of seven transovarial transmission was observed and the virus reisolated from larvae of first generation. However, the virus was not detected after nymphal metamorphosis. After blood feeding on viremic goats, 71% of the nymphae were infected. After metamorphoses 22% of the males and 42% of the females had a low virus titer. Rh. e. evertsi appears to have a limited efficacy in transmitting and replicating the CCHF virus but must be not neglected as a potential vector during an epizootic manifestation. PMID- 10472437 TI - [Heterogeneity of chloroquine resistant malaria in Senegal]. AB - In order to evaluate the in vivo efficacy of chloroquine in the treatment of P. falciparum malaria, studies have been carried out in Richard-Toll, Fatick and Tambacounda, 3 areas where dynamics of transmission, population movements, as well as prophylactic and curative practises are different. Failure rates in treatment were 13% in site 1 (Richard-Toll) where medical pressure and population movements are high. In sites 2 and 3 (Fatick and Tambacounda), the failure rate was 3%. Even if medical pressure is lower in the latter site (Tambacounda), the higher transmission could be a factor in the spreading of resistance. A surveillance system and better use of chloroquine must be undertaken. PMID- 10472438 TI - [Severe forms of malaria in children in a general hospital pediatric department in Yaounde, Cameroon]. AB - Severe forms of malaria in children are responsible for 1 million deaths yearly in young children in hyperendemic areas. The main objective of this study was to identify and compare common manifestations of different forms of severe malaria and to evaluate the prognosis for hospital treatment in an endemic area. 271 files of children admitted into hospital between March 1991 and September 1996 were analysed. These children were confirmed to have Plasmodium falciparum in their peripheral blood. 78 patients (29%) had the severe form of malaria. 43 patients (53%) were under 5 years of age. The 5 severe types identified were characterized by very high temperatures 28 cases (36%), cerebral malaria 20 cases (26%), prostration and weakness 15 cases (19%), severe anaemia 14 cases (18%) and haemoglobinuria 1 case (1.3%). Cerebral malaria and severe anaemia were more common in children under 5 years old. The average parasitemia was 16,366 +/- 1390 parasites per microlitre. Clearance of parasitemia was obtained on day 3 in almost all cases; 6 patients with very high temperatures presented neither sign of visceral complications nor convulsions. The average period in coma for cases of cerebral malaria was 1.7 days; 12 anaemic patients were transfused. There were no deaths. No abnormality was found on physical examination after an average hospitalisation of 5.3 days. An early diagnosis and adequate treatment of severe forms of malaria in children by qualified personnel will usually result in a favourable prognosis in our area. PMID- 10472439 TI - [Toxoplasmosis at the Pasteur Institute of Bangui, Central African Republic (1996 1998): serological data]. AB - A serological study of toxoplasmosis was conducted between 1996 and 1998 on 1953 patients of the Medical Analysis Laboratory of the Institut Pasteur de Bangui. The mean age of patients was 28 years. Among sera tested by ELISA, seropositivity to IgG antibodies was observed in 50.6%, and 2.6% sera were found positive for IgM antitoxoplasma antibodies (immuno-capture). The seroprevalence did not vary significantly according to sex or age. The results showed 40.8% sera had IgG antibodies titered 400 Ul/ml and more. The proportion of high level (> 400 Ul) IgG was more important in males than in females. High level IgG antibodies were statistically significant more frequently in the sera of females aged 10-29 years. Of the procreative women, 49.1% were at risk of contacting toxoplasmosis. The diagnosis of recently acquired infection, based on the coexistence of IgM antibodies and high level IgG antibodies, was noted in 1.6% among sera of procreative women. In the Central African Republic, serologic survey during pregnancy is not systematic and HIV seroprevalence is high (15%). Risks of acute infections during pregnancy and of opportunistic infections in HIV-infection patients are high. A control of toxoplasmosis is justifiable (screening during pregnancy, sanitary education) in CAR. PMID- 10472440 TI - [Hematologic abnormalities and immunodepression in HIV/AIDS- related pulmonary tuberculosis]. AB - Our objectives were to describe hemato-immunological abnormalities encountered in tuberculosis patients HIV seropositive (TBVIH+, n = 67) or not (TBVIH-, n = 39) and in HIV asymptomatic patients (aSVIH+, n = 40). We found: a great reduction of mean value of RBC and Hb in TBVIH+ and TBVIH-; a reduction of mean value of leucocytes, total lymphocytes and CD4+ lymphocytes in TBVIH+ and aSVIH+; an inversion of [formula: see text] ratio, more important in TBVIH+ than in aSVIH+. HIV and tuberculosis association, HIV1 serotype and CD4 < 200/mm3 were found to promote significantly hemato-immunological abnormalities. PMID- 10472441 TI - [Severe forms of eosinophilic meningitis in infants of Mayotte. Apropos of 3 cases]. AB - BACKGROUND: Eosinophilic meningitis caused by Angiostrongylus cantonensis is widespread in Southeast Asia and the Pacific islands. Adults develop transient meningitis with a benign course, whilst severe or fatal disease may occur in pediatric patients. CASE REPORTS: Three infant girls, aged 8 to 11 months, living on the island of Mayotte, developed fever, hypotonia, coma (2 cases), and, for one of them, seizures. Eosinophilia was detected in the peripheral blood and cerebrospinal fluid. Secondary, flaccid quadraplegia (1 case) or paraplegia (2 cases) with absence of deep tendon reflexes, urinary retention and anal incontinence were noted. Three patients had autonomic dysfunction. Computerized tomography showed enlarged ventricles and cerebral subarachnoid spaces. One patient had sequelae. Two patients could not be followed. Retrospectively, the diagnosis of angiostrongylus infection was established for two infants by a serological study. CONCLUSION: We report three new cases of infants with severe Angiostrongylus cantonensis infection in the French island of Mayotte (Comoro Islands). In this Indian Ocean area, eosinophilic meningitis seems to occur exclusively in infants and with severe radiculomyeloencephalitic forms. PMID- 10472442 TI - [Comparative analysis of 2 diagnostic methods of human loiasis: IgG4 serology and nested PCR]. AB - By evaluating the diagnostic methods developed in our laboratory, the prevalence of loaiosis was estimated among 201 individuals from the province of Haut Ogooue in Gabon using IgG4 serology and nested-PCR. The study showed that the prevalence of loaiosis was higher than that described using standard microscopy. IgG4-based ELISA (Enzyme Linked Immunosorbant Assay) using crude extract of Loa loa microfilariae showed that 80% (35/44) of microfilaraemic individuals (MF') and 56% (88/157) of amicrofilaraemics (AMF) presented antibodies. By contrast, L. loa specific DNA amplified by nested-PCR was detected in all MF and in 68% (106/157) of AMF. Among the 201 samples tested, 95 (47%) gave positive results in both tests. These results indicate that the presence of IgG4 antibodies directed against crude extract of L. loa microfilariae is not linked to the positivity of nested-PCR assay (chi 2 for paired data = 8.78; P < 0.02). We conclude that the PCR assay is more sensitive than the detection of IgG4 antibodies (directed against crude extract of L. loa microfilariae) in detecting loaiosis, and particularly occult loaiosis (infection without circulating microfilariae). PMID- 10472443 TI - [Therapeutic failure of melarsoprol among patients treated for late stage T.b. gambiense human African trypanosomiasis in Uganda]. AB - The failure rate of melarsoprol after treatment of late stage cases of Human African Trypanosomiasis (HAT) is usually under 7%, even though the drug has been used for such treatment over the past 50 years. We report a melarsoprol treatment failure rate of 26.9% among 428 patients treated in Northern Uganda. Whatever its origin, this observation, the first documented in a HAT focus, is alarming, particularly since no second line trypanocidal drug is actually available for the treatment of late stage HAT. We believe that the current worrisome situation of HAT in several African countries and the risk of emergence of other foci of resistance, argue in favour of a greater attention on the part of the scientific community and the pharmaceutical companies being paid to this problem. PMID- 10472444 TI - [Paracoccidioides brasiliensis infection in a subtropical region with important environmental changes]. AB - Paracoccidioidomycosis is a systemic mycoses caused by Paracoccidioides brasiliensis, a dimorphic fungus that infects man through respiratory ducts. It has been accepted that its ecological niche is located in the soil and plants of subtropical forests of Latin America. The Province of Corrientes is located at the northeastern border of Argentina, in a subtropical area where important environmental modifications have been introduced in the last decade as consequence of damming the Parana river at Yacyreta, one of the biggest hydroelectric dams in the world. Since there are no data on human infection provoqued by this fungal agent in Corrientes, the purpose of this study was to obtain information at present time about infection indexes and to assess if environmental changes introduced in the area could impact on the epidemiology of the disease. Skin tests with paracoccidioidin and histoplasmin were performed on 455 persons of both sexes, from 1 to 73 years of age, who live permanently in the area and voluntarily accepted to be included in the study. Both antigens were employed at the same time in order to evaluate crossed type reactions. Of the 455 persons, 52 (39 males-13 females) were reactive to paracoccidioidin (11.4%), with an increasing prevalence with age. According to previous data, these results would indicate an increase in the index of human infection by P. brasiliensis, and this may be related to the important changes in climatic and environmental conditions introduced in the area in the last years. PMID- 10472445 TI - [Epidemiological stratification of malaria in the Comoro archipelago]. AB - In the Comoros Islands, the level of malarial endemicity varies greatly from one island to the other, even though the total area (4 islands) covers less than 2,300 km2 and has a population of some 600,000 people only. The epidemiological stratification is based on the diversity of human and physical characterisation. They both determine the presence and the behaviour as well as the size of the vector's populations. Vectorial dynamics can explain varying levels of endemicity given parasitological indicators and specific morbidity. Analyzing these criteria shows up different epidemiological features and serves as a basic guideline for malaria control. The efficiency of this control depends on the relationships between the intensity and the length of the transmission, in the framework of protection mechanisms; it is of crucial importance for clinical treatment. Further elements are the age of the patient, the season and the geographic situation of the area. Stratification provides explanations for these relationships and helps to define antimalarial programmes adapting to each situation a range of therapeutic and antivectorial methods. The availability and accessibility of anti-malarial medicine is the minimum requirement for reducing mortality: domestic spray insecticides for reducing transmission are effective for several years and should be followed by the use of mosquito nets or curtains impregnated with pyrethrinoids, and in the particular case of Grande Comore, the use of larvivorous fish. As anywhere else, the economic development, which is dependent on political stability, is the essential basis for malaria control. PMID- 10472446 TI - [The focus of human trypanosomiasis in Campo (Cameroon). History and endemic situation in 1998]. AB - For the first time in the last thirteen years, the human sleeping sickness focus at Campo, spanning the Cameroon-Equatorial Guinea border areas, has been prospected. The screening was carried out simultaneously on both sides of the border. This focus has been known since the beginning of the century but, contrary to what took place in other well-known foci in bordering countries south of Cameroon, either in the 1920s or the 1980s--there has never been an epidemic outbreak in that area. Such an epidemiological situation makes this focus particularly interesting. Though still active, trypanosomiasis is not very manifest. According to passive screening carried out in recent years, the estimated prevalence ranges between 0.2 and 0.5%. For this screening, 5,255 persons were examined on the Cameroonian side of the focus (90.6% of the census population). The serological screenings were carried out with the CATT 1.3, which is the CATT generally used in screening, and with the latex CATT which associates LiTat 1.3, 1.5 and 1.6. The search for trypanosomes was made by testing the lymph nod juice in presence of adenopathy and in the blood by Quantitative Buffy Coat (QBC), the mini anion exchange centrifugation (mAEC), as well as the in vitro culture using the kit for in vitro isolation of trypanosomes (KIVI) for individuals suspected to be serologically positive. 16 patients were identified in Cameroon but none in Equatorial Guinea. The results show that the Campo focus is active only on the Cameroonian side, centred on the village of Ipono with a limited prevalence (0.3%). The persisting epidemic is most likely to be associated with the presence of pigs carrying the Trypanosoma brucei gambiense which was identified during the study in Ipono. The strain that we isolated was studied by isoenzyme electrophoresis on cellulose acetate. Its zymodeme is the same as that of the human strain isolated in Campo. With the collected epidemiological data, a concerted medical and entomological action could be planned within the limits of the village of Ipono to eradicate the disease. This action may be organised by the existing local health structures. During this study, the latex CATT proved to be more cost-effective than the CATT 1.3 since a similar result was reached requiring eight times less work at a lower cost. This remains to be confirmed in a hyperendemic focus. PMID- 10472447 TI - [Epidemiological observations on the first case of human paragonimiasis and potential intermediate hosts of Paragonimus sp. in Benin]. AB - Parasitological investigations were carried out for four months in 1998 in two Beninese centres of pneumo-phtisiology (Akpakpa, at Cotonou, and Akron, at Porto Novo) to detect the patients harbouring eggs of Paragonimus sp. amongst the persons consulting for tuberculosis and showing a broncho-pneumopathy without mycobacteria. Eggs of Paragonimus sp. were detected in the sputum of a single patient out of 369 persons examined (prevalence, 0.2%). This patient had eaten crabs in the months preceding the date of diagnosis. A treatment using praziquantel has improved clinical symptomatology and biological signs found in this patient. These studies have been completed by visiting markets located in the coastal plain of Benin to identify the crabs that were sold and to find metacercariae of Paragonimus sp. Negative results were obtained when 126 Cardisoma armatum ("hole crab") were dissected. In contrast, the dissection of 176 Callinectes marginatus ("swimming crab") was successful, with 5% of crabs harbouring metacercariae of probably Paragonimus sp. Further studies are necessary to confirm these first results and to determine the global prevalence of Paragonimus infection in these definitive and intermediary hosts. PMID- 10472448 TI - [Schistosomiasis in the region of Ziga (Burkina Faso) before the construction of a dam]. AB - The authors report the results of a survey on schistosomiasis in the zone of Ziga (Burkina Faso) where a dam is going to be built. They examined 438 children aged from 7 to 15 years in 5 villages. The appraisal of macroscopic or microscopic hematuria, urine filtration, stool analysis and ultrasonographic examination made possible the assessment of the prevalence and severity of both schistosomiasis. The area was hyperendemic for urinary schistosomiasis. The general prevalence of S. haematobium was higher than 46% and 55% of the children presented at least one lesion of the bladder and 2% of hydronephrosis at ultrasonography. Intestinal schistosomiasis was not observed although few cases have been described in the regional hospital and intermediate host snails have been collected in the area. A surveillance of schistosomiasis, based on school surveys, was strongly recommended in the dam area. PMID- 10472449 TI - [ The aggressive cycle of Anopheles gambiae s.s. inside houses and malaria transmission in the Bouake region (Cote d'Ivoire). Value of using impregnated mosquito nets]. AB - The biting indoor cycle of Anopheles gambiae s.s. was studied in a rural zone near Bouake from 6 pm to 6 am. The cycle was characterized by a peak between midnight and 1 am (n = 19,181). The parturity rate of biting females regularly increased from 6 pm until 3 am, then remained at a high level up to 6 am (n = 14,239). Infected bites were observed from 7 pm to 6 am with a maximum between 0 and 3 am, and 96.7% of them were obtained between 11 pm and 4 am (n = 219). It appears that malaria transmission occurs essentially at the moment when villagers are in bed. The correct use of impregnated bed nets may thus greatly reduce malaria transmission. PMID- 10472450 TI - [An experimental model to analyse the characteristics of a strain of Culex pipiens and its sensitivity to insecticides]. AB - This study, based on phenotype of green larvae colour, has enabled the selection of a genetically distinct line of Culex pipiens pipiens. The four loci under study were found to be homozygous to the 42nd generation. These loci were associated with an increased sensitivity to organochlorides in the progeny when compared to the parent strain. This observation, with possible expansion to other species, has potential practical applications in the identification of susceptible insect populations in insecticide campaigns. PMID- 10472451 TI - [Evaluation of the vectorial capacity of Anopheles arabiensis (Diptera:Culicidae) on the island of Reunion: an approach to the health risk of malaria importation in an area of eradication]. AB - Following intensive control measures, malaria was eradicated from La Reunion Island (Indian Ocean) in 1979. However, potential vectors remain in coastal areas and, each year Public Health Services detect some imported cases from surrounding countries. Anopheles arabiensis is the only species which can be responsible for local transmission though its brief life cycle and its exophilic and zoophilic behaviour were found to be key elements in maintaining the eradication status of the disease on the island. Its vectorial capacity was measured at different seasons and in different areas with a view to assessing the health risks due to imported malaria and suggesting a more pertinent strategy of vector control. PMID- 10472452 TI - [Age group distribution of Glossina palpalis palpalis females in plantations and shallow water of the forest region of Cote d'Ivoire. Relation to the prevalence of sleeping sickness]. AB - In epidemiologically dangerous biotopes of foci with high prevalence of Human African Trypanosomiasis (HAT), different female age groups (nulliparous, young parous, old parous) were observed in the same proportions. On the contrary, in areas without HAT or in low prevalence foci (< 0.2%), these proportions significantly differed. Female age group distribution in epidemiologically dangerous biotopes could thus be a good indication by which to guide the control of Human African Trypanosomiasis (HAT). PMID- 10472453 TI - Day case tonsillectomy: what is the risk and where is the economic benefit? PMID- 10472454 TI - Posterior tragal perichondrium harvesting for myringoplasty. AB - We present a modified harvesting approach for tragal perichondrium, used in tympanic membrane reconstruction. The technique described avoids amputation of the targus thereby facilitating dissection of the perichondrium from the cartilage as compared to the traditional method. The approach described is technically easier, and removes any potential for cosmetic deformity associated with tragal cartilage amputation and reimplantation. Furthermore, both the anterior tragal perichondrium and the temporalis fascia remain intact if further surgery is required. We recommend this approach for permeatal, tragal perichondrial grafting of small to medium sized tympanic membrane perforations. PMID- 10472455 TI - Clinical risk management: principles of consent and patient information. AB - Clinical Risk Management and its integral elements form part of the foundation of 'Clinical Governance'. An important principle within Clinical Risk Management is that of consent and patient information which, in a changing climate of complaint and litigation, means that it is essential that clinicians have a sound understanding of its importance. This paper provides a structured understanding of the background and practice of consent and patient information. With such knowledge clinicians can help patients achieve a better understanding of what they are undertaking, and so reduce the risk of complaint and litigation. This paper discusses the topics of: current indemnification, ethics, law, the information content required for consent, duration of consent, who should take consent, capacity to give consent, mental incapacity, and details of how to perform risk assessment. However, areas applicable to the Mental Health Act have been excluded. PMID- 10472456 TI - The anatomy of the lacrimal bone at the lateral wall of the nose: its significance to the lacrimal surgeon. AB - The position, dimension and thickness of the exposed lacrimal bone at the lateral nasal wall in 10 cadaveric half-heads were examined. In all cases, the lacrimal bone at the lateral nasal wall was found to be just anterior to the mid-third of the uncinate process. The average length and width was 7.4 mm and 2.5 mm, respectively. In nine of the 10 half-heads, the lacrimal bone was very thin with an average thickness of 57 mm. In all the cases, the position of the lacrimal passage covered by the lacrimal bone corresponded to the postero-medial aspect of the upper lacrimal duct and the lower lacrimal sac. This study shows that the uncinate process is a reliable landmark for the lacrimal bone in endoscopic nasal surgery. The paper-thin lacrimal bone allows a bone rongeur to infracture through and nibble away the bony covering of the lacrimal sac in a dacryocystorhinostomy. PMID- 10472457 TI - Dry tap at myringotomy: a three-year study of 1688 children undergoing myringotomy. AB - One thousand six hundred and eighty-eight children undergoing myringotomy were studied to determine the rate of 'dry taps' and factors associated with a dry middle ear at operation. The percentage of bilateral dry taps was 16%. The dry tap rate was significantly higher in the following groups: older children, those with an A, C1 or C2 tympanogram at the time of listing, milder hearing loss, a preceding history of recurrent acute otitis media and operation performed during the summer or autumn. There was no significant relationship between the dry tap rate and the time on the waiting list, the gender of the child or the order in which right or left myringotomies were performed. Careful preoperative reassessment of any child with a type A, C1 or C2 tympanogram should be made to prevent unnecessary surgery. Separating those listed for treatment of recurrent acute otitis media from those with OME would prevent these being included in the overall dry tap rate. PMID- 10472458 TI - Squamous cell carcinoma antigen in patients with cancer of the larynx. AB - The purpose of this study was to evaluate the clinical usefulness of squamous cell carcinoma antigen (SCC Ag) in patients with squamous cell carcinoma of the larynx. Plasma specimens were obtained from 70 patients with cancer of the larynx before and after treatment and during follow-up. Disease status and the marker levels were determined blind to each other. Microparticle enzyme immunoassay (IMx SCC) was used to measure the SCC Ag level. Applying standard normal limits the sensitivity of the marker at diagnosis was 25.7%. SCC Ag levels were generally lower after therapy than before. Relapse occurred more often in patients with an abnormal pretreatment SCC Ag level, which was more frequent in those with nodal invasion. The marker level increased in 70% of the patients with relapse before the clinical detection of recurrence. SCC Ag is of limited usefulness in the primary diagnosis of cancer of the larynx, but is useful in detecting recurrence of cancer. PMID- 10472459 TI - An assessment of the value of the preoperative computed tomography scans prior to otoendoscopic 'second look' in intact canal wall mastoid surgery. AB - 'Second look' surgery following primary intact canal wall mastoid surgery for cholesteatoma is considered mandatory for most cases in modern otological practice. The morbidity of the second look can be reduced by the use of the rigid otoendoscope. Forty-three patients undergoing 'second look' surgery were studied with an average age of 24.7 years. Prior to surgery a computed tomography (CT) scan was performed to assess the anatomy and pneumatisation of the cavity. The mean interval between primary and secondary surgery was 16 months and in all cases CT scans were performed within 6 months of 'second look' surgery. The presence of an opaque mastoid did not correlate with residual or recurrent cholesteatoma. The sensitivity of CT in diagnosing residual or recurrent cholesteatoma was 42.9% with a specificity of 48.3% and a predictive value of 28.6%. These results are explained by the fact that it is radiologically impossible to differentiate between recurrence, scar tissue or fluid with a CT scan. Nevertheless it was possible to inspect the cavity with the otoendoscope even in the presence of an opaque mastoid whether due to scar tissue or residual/recurrent cholesteatoma. PMID- 10472460 TI - Angular cheilitis after tonsillectomy. AB - The operation of tonsillectomy requires the oral cavity to be held open mechanically in an unconscious patient, and intra-oral instrumentation to occur. Angular cheilitis may arise as a result of this after operation. This can cause morbidity and delay the re-establishment of a normal diet. The aim of this study was to identify what factors increase the likelihood of developing this problem postoperatively. Sixty patients were randomly selected in a prospective manner. Preoperative, intraoperative and postoperative variables were recorded. The frequency of development of postoperative angular cheilitis was recorded. The prevalence of the condition was related to the prerecorded variables. Parametric analysis showed that the chance of developing angular cheilitis was directly related to the use of diathermy haemostasis (P = 0.05). Logistic regression analysis showed that the odds ratio of developing this complication if diathermy was used is 3.5 (95% confidence intervals 0.99, 12.4) and operation difficulty may also be a relevant variable. No other recorded variables were found to be significant. PMID- 10472461 TI - Apoptosis in meatal skin, cholesteatoma and squamous cell carcinoma of the ear. AB - Apoptosis in normal meatal skin, cholesteatoma and squamous cell carcinoma (SCC) of the ear was investigated by using morphological analysis and in-situ specific labelling of fragmented DNA. In meatal skin, apoptotic cells were localized mainly in the granular layers, but were not so restricted in cholesteatoma, while in SCC they were even more dispersed. The apoptotic index (AI) was low (1.59 +/- 0.10 SEM) in normal skin. It was increased in cholesteatoma (2.09 +/- 0.11), and was intermediate in SCC (1.72 +/- 0.14). By contrast, the mitotic index (MI) increased from 0.19 +/- 0.02 in normal skin, to 0.25 +/- 0.01 in cholesteatoma and to 0.25 +/- 0.02 in SCC. Our findings indicate that apoptosis is involved in the epithelial homeostasis of meatal skin, cholesteatoma and SCC of the ear. The hyperproliferation of epithelial cells in cholesteatoma is counteracted by an increased apoptosis rate, while in SCC the increased cell proliferation without a compensatory increase in apoptosis may be associated with the malignant transformation. PMID- 10472462 TI - Dizziness symptom severity and impact on daily living as perceived by patients suffering from peripheral vestibular disorder. AB - The purpose of this study was to describe quality-of-life aspects in a group of patients (n = 99) suffering from peripheral vestibular disorder, using three different self-rated instruments, the Vertigo Symptom Scale (VSS), the Vertigo Handicap Questionnaire (VHQ) and the Sickness Impact Profile (SIP). The results showed that the type of dizziness that most influenced the quality-of-life aspects were: frequent short- or long-term dizziness, nausea, and the feeling that the ground was distant or as though the patient were walking on clouds. However, several of the impairments in daily life were neither related to the disease itself nor the demographic data. This verifies the necessity of investigating other factors such as personality and coping capacity. The results of this study also demonstrate the patients' need of psychosocial support. A comprehensive assessment and evaluation is important in order to identify each patient's needs. PMID- 10472463 TI - Sensitivity, specificity and predictive value of tympanometry in predicting a hearing impairment in otitis media with effusion. MRC Multi-Centre Otitis Media Study Group. AB - Children with otitis media with effusion (OME) need monitoring over time to identify persistence of disease and to assess their hearing thresholds as a surrogate of auditory disability. It would be useful if tympanometry could be used to predict those with an impairment. This study looked at 1153 children, aged between 3.25 and 6.75 years, referred for suspected OME. The inclusion of type C2 tympanograms with type B tympanograms versus all other tympanogram types increased the sensitivity of detecting air-conduction thresholds and air-bone gaps of various magnitudes in the better hearing ear but lowered the specificity and the positive predictive value. Extending the pressure range to -600 daPa and excluding the 30% of children with poor concentration on audiometry made no difference to the sensitivity and specificity. The results were the same for the poorer hearing ear. These findings are of practical help in monitoring children with OME. Thus taking an air-conduction average of 25 dB HL in the better ear as the level needing detection, if all children are audiometrically assessed then 100% of those with an impairment will be identified. Limiting audiometry to those with a bilateral type B tympanogram reduces the workload by 50%, but 90% of impaired children will still be detected. Limiting audiometry to those with type B or C2 tympanograms reduces the workload to 69% of the sample, and 95% of impaired children will be identified. With such data, decisions as to how to allocate limited audiometric resources for monitoring children with OME are made easier. PMID- 10472464 TI - The large vestibular aqueduct syndrome: the role of cochlear implantation in its management. AB - In recent years the large vestibular aqueduct syndrome has become an increasingly recognized cause of a progressive sensorineural hearing loss. Cochlear implantation, although not correcting the bony abnormality, does offer an avenue for rehabilitation for affected individuals and initial results are encouraging. Of 15 patients identified with the large vestibular aqueduct syndrome we have implanted seven (five adults and two children). All patients underwent an uneventful electrode insertion with uncomplicated postoperative periods. Three of our adult patients are showing very good initial results with BKB speech recognition scores of 100%, 74% and 66% with the implant alone. It is still early days with the children, but initial results are encouraging. A full review of the large vestibular aqueduct syndrome is given along with our results. PMID- 10472465 TI - Nasal congestion during pregnancy. AB - We define pregnancy rhinitis as nasal congestion in the last 6 or more weeks of pregnancy without other signs of respiratory tract infection and with no known allergic cause, disappearing completely within 2 weeks after delivery. In order to describe physiological variations of nasal obstruction during pregnancy, subjective scores and nasal as well as oral peak expiratory flow values were recorded daily in 23 pregnancies until 1 month after delivery. Scores were higher during early and late pregnancy than in the month after delivery. Objectively registered blockage increased during pregnancy in eight women only. Unexpectedly nine women showed declining blockage. Five of 23 women had pregnancy rhinitis. PMID- 10472466 TI - The effect of pH of douching solutions on mucociliary clearance. AB - The effect of the pH of hypertonic saline nasal douching solutions on mucociliary clearance was studied in order to investigate the common notion that such solutions should be buffered alkaline. Thirty normal subjects were included in a randomised controlled crossover trial. Mucociliary clearance was measured by the saccharin clearance time. There was no difference in mucociliary clearance after douching with a non-buffered solution and a solution buffered to pH 8. However both solutions significantly improved mucociliary clearance compared to the baseline (P < 0.001) probably on account of their hypertonicity. PMID- 10472467 TI - Long-term results of posterior semicircular canal occlusion for intractable benign paroxysmal positional vertigo. AB - Benign paroxysmal positional vertigo (BPPV) is a common condition which is usually managed conservatively, surgical intervention being recommended only for those small number of patients in whom it becomes persistent and incapacitating. The results of surgery in 13 patients who underwent posterior semicircular canal occlusion for intractable (> 12 months duration) and incapacitating BPPV are presented with special emphasis on their long-term follow-up. The mean follow-up was 66 months (range, 29-119 months). All patients reported complete and immediate resolution of their positional vertigo, which has been maintained in the long term. Most patients, however, reported some postoperative transient unsteadiness which lasted up to 4 weeks. All patients developed a transient mild conductive hearing loss secondary to a middle ear collection, which usually resolved within 4 weeks. Five patients developed a transient mild high frequency sensorineural hearing loss which resolved in all cases within 6 months. There were no reports of sensorineural hearing loss nor tinnitus in the long term. All patients believed that the operation was beneficial and would undergo it again. Our findings indicate that posterior semicircular canal occlusion is an effective and safe operation in the long term and is the procedure of choice for intractable and incapacitating BPPV rather than singular neurectomy. PMID- 10472468 TI - The subjective effects of smoking on nasal symptoms. AB - One hundred and forty-four volunteers completed a validated questionnaire, to investigate the association between smoking, non-smoking and passive smoking status, and the degree of a range of 10 common nasal symptoms. Logistic regression showed that smokers were four times and passive smokers six times more likely to report hyposmia compared to non-smokers. No significant differences were found between the three groups for any of the other nine symptoms. PMID- 10472469 TI - Imaging of temporal bone brain hernias: atypical appearances on magnetic resonance imaging. AB - Brain tissue with or without its dural covering may protrude into the temporal bone through a defect in the tegmen tympani. Infection or granulation tissue, with or without cholesteatoma may make diagnosis difficult. While computed tomography (CT) may demonstrate a bony defect, it is difficult to distinguish between herniated brain, cholesteatoma or granulation tissue. Magnetic resonance imaging (MRI) clearly demonstrates healthy herniated brain tissue, but ischaemic or necrotic elements may mimic other lesions. We present a series of five patients with brain hernia, highlighting the spectrum of preoperative radiological appearances, and some of the difficulties encountered in interpreting these images. MRI demonstrated herniation of healthy brain in two patients but in three, showed irregular soft tissue with signal characteristics consistent with cholesteatoma. Surgery in these three patients demonstrated necrotic brain with coexisting cholesteatoma in two. Brain hernias are often necrotic and may have morphological and signal characteristics similar to cholesteatoma with which they may coexist. PMID- 10472471 TI - Psychometric performance before and after laser palatoplasty for non-apnoeic snoring. AB - Psychometric tests are an objective way of examining cognitive functioning, and have shown impairment in patients with obstructive sleep apnoea. Non-apnoeic snoring may cause reduced concentration, but psychometric tests have been used rarely in this population. We investigate whether their use can demonstrate an improvement in cognitive performance in 20 non-apnoeic snorers following Laser Palatoplasty (LPP). The subjects completed psychometric tests, Beck Anxiety and Depression Inventories and an Epworth Sleepiness Scale on two occasions before LPP and once postoperatively. The only index to show any significant change with LPP was the Beck Depression Inventory (P < 0.005), which was reduced by a mean of 1.6 units following surgery. There was also a highly significant improvement in information processing between the preoperative tests, for which no explanation could be found. Our results suggest that commonly applied psychometric tests are unable to demonstrate significant improvements following surgery for non-apnoeic snoring. However, we have demonstrated for the first time a significant reduction in depression following surgery, which is evidence that snoring is more than a social nuisance. PMID- 10472470 TI - Objective benefit of laser palatoplasty for non-apnoeic snoring. AB - Laser palatoplasty (LPP) is widely used for the treatment of non-apnoeic snoring, despite the lack of objective data supporting its use. We report measurements of snoring in a prospective study of LPP, and we compare the results with a previous study of uvulopalatopharyngoplasty (UPPP). Twenty patients with an apnoea/hypopnoea index < 20 h-1 underwent LPP for habitual snoring. Overnight sound recordings were compared before and 6 months after operation using three objective indices; L, (the level exceeded by the loudest 1% of sound), L5 (the level exceeded by the loudest 5% of sound) and P50 (% total sleep time above 50 dBA). The subjective impression of snoring severity (Wilcoxon test, P < 0.001), and objective indices L1 and P50 (t-test, P < 0.001) showed significant reductions after LPP. The mean change in L1 was 4.2 dBA, comparable to that we previously reported for UPPP, while P50 was reduced to less than one-third its preoperative value. No other sleep variables changed significantly following LPP. We conclude that LPP results in reduced snoring volume comparable to that following UPPP. PMID- 10472472 TI - Risk factors for chronicity in childhood otitis media with effusion. AB - One hundred and ninety-one children who presented to the author's clinic with otitis media with effusion (OME) over a 2-year period (1986-1988) have been studied prospectively. Resolution of their OME was considered to have occurred when they had normal findings on pneumatic otoscopy, tympanometry and audiometry on at least two occasions 6 months apart and had subjectively normal hearing in between. The number of operations performed during the course of the disease was used as a surrogate for chronicity. Children with a history of previous aural discharge at presentation were significantly more likely to have chronic OME (P = < 0.02). PMID- 10472473 TI - The low-power laser in the treatment of tinnitus. AB - The purpose of this study was to evaluate the low-power laser on the treatment of tinnitus. In a randomized, prospective, double-blind, placebo-controlled trial, either active or placebo low-power laser irradiation was given through the external acoustic meatus of the affected ear towards the cochlea. The active laser applied 50 mW (cw, 830 nm) over a period of 10 min per session. Forty-nine patients with severe, chronic uni- or bilateral tinnitus were studied. The main outcome was measured using psychoacoustical match of tinnitus loudness and pitch, Visual Analogue Scale (VAS) ratings of subjective loudness, annoyance and attention involved, scores on the Tinnitus Handicap Inventory (THI), the Tinnitus Coping Style Questionnaire (TCSQ), and a number of psychosocial questionnaires. The results showed only moderate (18%) subjective improvement with no statistically significant differences between the effects of the active laser and placebo treatments. Also, there were no statistically significant differences in prepost measurements of tinnitus loudness, VAS scores, THI scores, or TCSQ scores for patients treated with active laser compared with those treated with placebo. We conclude that low-power laser treatment is not indicated in the treatment of tinnitus. Reports of significant benefits of this treatment in previous, mostly uncontrolled or single-blinded studies may be explained by the placebo effect. PMID- 10472474 TI - Classification and staging of cholesteatoma. AB - No staging system for cholesteatoma has been universally adopted. All previous attempts failed to gain acceptance because of the lack of clinical relevance. We have devised a simple system to stage cholesteatoma which is based on the extent of lesion, ossicular condition and the preoperative complications. To illustrate the systems potential value, data from 119 new patients with cholesteatoma are presented and classified according to our proposed scheme. This shows a relationship between the stage of disease, ossicular damage and the occurrence of complications. The proposed staging system is potentially useful when considering the type of surgery required and when comparing published data reporting the results of tympanomastoid surgery. PMID- 10472475 TI - A study of postoperative fever following paediatric tonsillectomy. AB - The aetiology of post-tonsillectomy fever is obscure. Bacteraemia during surgery, anaesthetic agents and the inflammatory response of tissue to injury have been implicated. A prospective study was undertaken in 100 consecutive children to evaluate the occurrence and severity of fever in the 24 h after tonsillectomy and its relationship to bacteraemia during surgery and qualitative and quantitative cultures (colony counts) of organisms in tonsil core tissue. Fifty-four patients had a fever (> 37.5 degrees C) postoperatively, of whom, 30 had a fever greater than 38 degrees C. Blood cultures during tonsillectomy were positive in 22 patients. There was no statistically significant difference between the occurrence of fever and the techniques of tonsillectomy and haemostasis. There was also no association between positive blood, core or surface cultures and the incidence or severity of fever nor any association between colony count in core cultures and fever. Our results suggest that postoperative fever in the 24 h following tonsillectomy is not caused by infection. PMID- 10472476 TI - Frey's syndrome following parotidectomy: prevention using a rotation sternocleidomastoid muscle flap. AB - The benefit of using a sternocleidomastoid flap following parotidectomy to reduce the incidence of symptomatic gustatory sweating (Frey's syndrome) was reviewed. A retrospective study was undertaken to review subjective and objective symptoms of Frey's syndrome in two groups of patients, one of whom had undergone superficial parotidectomy with a sternocleidomastoid flap rotated at the time of surgery and a second group of patients who had undergone a standard superficial parotidectomy. A total of 22 patients, randomly sampled and willing to attend, were evaluated postoperatively, at a median time from surgery of 42 months (range 23-82 months) in the non-flap group and 44 months (range 14-66 months) in the flap group, by Minor's starch iodine test. In the 11 patients who had a sternocleidomastoid flap rotated, two had evidence of gustatory sweating. Of the 11 that had not undergone sternocleidomastoid flap rotation, nine patients showed evidence of gustatory sweating (P < 0.05, chi 2 test). There were two patients in total who had clinical symptoms of Frey's syndrome and both of these had not undergone flap rotation at the time of parotidectomy. PMID- 10472477 TI - FUO and AIDS. PMID- 10472478 TI - Cytomegalovirus: disease syndromes and treatment. PMID- 10472479 TI - Diagnosis of respiratory viruses in cancer and transplant patients. PMID- 10472480 TI - Sepsis in obstetric and gynecologic patients. PMID- 10472481 TI - Emerging nosocomial infections and antimicrobial resistance. PMID- 10472483 TI - Hospitalizing influenza in adults. PMID- 10472482 TI - Current concepts on animal bites: bacteriology and therapy. PMID- 10472484 TI - Chronic fatigue syndrome: current concepts of pathogenesis and treatment. PMID- 10472485 TI - Current considerations in the management of fever and neutropenia. PMID- 10472487 TI - Rhodococcus equi infections: clinical features and laboratory diagnosis. PMID- 10472486 TI - Pneumonia, tuberculosis, and urinary tract infections in pregnancy. PMID- 10472488 TI - Managed health care: outcomes and update. PMID- 10472490 TI - Human immunodeficiency virus disease in the era of HAART: a reevaluation of the mucocutaneous manifestations. PMID- 10472489 TI - Prions and prion diseases of the central nervous system. PMID- 10472491 TI - Xanthogranulomatous pyelonephritis. PMID- 10472492 TI - Current diagnosis and management of chronic fungal infection of the feet and nails. PMID- 10472493 TI - Topical retinoids in the treatment of acne vulgaris. PMID- 10472494 TI - Parental marital disruption and intergenerational transfers: an analysis of lone elderly parents and their children. AB - Although one of the most marked demographic trends observed over the twentieth century is the increased rate of divorce, relatively little research has explored the effects of these changing marital patterns in the context of an aging society. Using a sample of lone elderly parents and their adult children, we analyze the direct and indirect effects of marital disruption on four important dimensions of intergenerational transfers: coresidence, financial assistance, adult children's provision of informal care, and parental purchase of paid care. Our findings suggest that divorce has deleterious effects on intergenerational transfers, particularly for elderly fathers. Remarriage further reduces exchange. Our results reveal that parents engage in lower levels of transfers with stepchildren relative to biological children. Moreover, intergenerational transfers are sensitive to characteristics of biological children but not to those of stepchildren. Taken together, these results suggest that exchange at the end of the life course continues to be adversely affected by marital disruption. PMID- 10472495 TI - Have family income mobility patterns changed? AB - We examine the mobility of individuals in the United States based on equivalent family income--that is, total income of all family members adjusted for family size according to the equivalence scale implicit in the U.S. poverty line. Our analysis, which tracks movements across quintiles, centers on four questions: How much movement is there across the family income distribution? How has this mobility changed over time? To what extent are the movements attributable to factors related to changes in family composition versus events in the labor markets? In light of major socioeconomic changes occurring in the quarter-century under study, have the determinants of mobility changed over time? Our findings indicate that mobility rates in the 1980s differed little from those in the 1970s. However, individuals in families headed by a young person or a person without a college education were less likely to experience upward mobility in the 1980s than in the 1970s. PMID- 10472496 TI - Shifting family definitions: the effect of cohabitation and other nonfamily household relationships on measures of poverty. AB - The current official poverty measure compares income to needs within a family. Some have suggested including cohabiting couples as part of this family. Others have suggested that the household be used as the unit of analysis for poverty measurement. I explore issues involved in expanding the unit of analysis, including the stability of cohabiting and other nonfamily household relationships and the degree of resource sharing that takes place among different types of people within households. Instability in households with nonfamily members is not a serious problem for inferring poverty from cross-sectional studies. On the other hand, income from people in nonfamily household roles contributes slightly less to helping other household members avoid financial hardship, implying that nonfamily housemates have a greater tendency to keep income to themselves. PMID- 10472497 TI - Trajectories of fetal loss in the Czech Republic. AB - Using data for 555,038 pregnancies conceived in the Czech Republic in 1987-1990, we show that pronounced differences in fetal survival in the middle trimester of pregnancy by marital status, educational level, and labor force attachment become much smaller at full term; survival differences by age at conception and number of previous deliveries show relatively constant proportional hazards throughout gestation. Social inequalities in postpartum life chances have been documented previously, but we show that similar inequalities exist before birth. PMID- 10472498 TI - Migradollars and mortality: the effects of migration on infant survival in Mexico. AB - We apply multilevel methods to data from Mexico to examine how village migration patterns affect infant survival outcomes in origins. We argue that migration is a cumulative process with varying health effects at different stages of its progression, and test several related hypotheses. Findings suggest higher rates of infant mortality in communities experiencing intense U.S. migration. However, two factors diminish the disruptive effects of migration: migradollars, or migrant remittances to villages, and the institutionalization of migration over time. Mortality risks are low when remittances are high and decrease as migration becomes increasingly salient to livelihoods of communities. Together, the findings indicate eventual benefits to all infants, irrespective of household migration experience, as a result of the development of social and economic processes related to U.S. migration. PMID- 10472499 TI - The impact of specific occupation on mortality in the U.S. National Longitudinal Mortality Study. AB - We compare mortality differences for specific and general categories of occupations using a national cohort of approximately 380,000 persons aged 25-64 from the U.S. National Longitudinal Mortality Study. Based on comparisons of relative risk obtained from Cox proportional-hazards model analyses, higher risk is observed in moving across the occupational spectrum from the technical, highly skilled occupations to less-skilled and generally more labor-intensive occupations. Mortality differences obtained for social status groups of specific occupations are almost completely accounted for by adjustments for income and education. Important differences are shown to exist for selected specific occupations beyond those accounted for by social status, income, and education. High-risk specific occupations include taxi drivers, cooks, longshoremen, and transportation operatives. Low-risk specific occupations include lawyers, natural scientists, teachers, farmers, and a variety of engineers. PMID- 10472500 TI - Entry or exit? A transition-probability approach to explaining the high prevalence of single motherhood among black women. AB - I analyze the prevalence of single motherhood among black and non-Hispanic white women in terms of differences in entry and exit. Higher initial entry rates among black women, especially through unpartnered childbearing, account for slightly more than half the difference between blacks and whites in the prevalence of single motherhood. The remainder of the difference is due to black single mothers' much lower rates of exit through union formation and to their very high rates of reentry through dissolution of these later unions. Entry and exit rates through the 1990s imply a widening racial gap. PMID- 10472502 TI - A century of U.S. censuses and the language characteristics of immigrants. AB - Since 1890, every U.S. census but one has asked about the language characteristics of the U.S. population. This almost uninterrupted data series, however, has been shaped by contemporaneous presumptions about the ties between language and ethnicity, the likelihood of proficiency in English among various subgroups, and practical constraints. I describe shifts across censuses in the phrasing of questions about language, the coding of responses, and the subpopulations for which the questions were asked and the results were published. I then describe the data generated by these items and discuss their interpretation. I conclude with a summary of the major insights and limitations of a century's worth of data. PMID- 10472501 TI - Time spent in parenthood status among adults in the United States. AB - Transition rates estimated from the 1987-1988 and 1992-1994 waves of the National Survey of Families and Households imply that a U.S. adult will spend approximately one third of the years from ages 20 to 69 as a parent of a dependent child. I distinguish biological from social parenthood and provide separate estimates by gender and race. White women conform most to the conventional image of a biological parent residing with an own child, whereas African American women spend the most adult years as a parent. On average, white men spend fewer years as parents than African American men, but African American men spend more years as biological parents not residing with any children. Implications of these descriptive findings are discussed. PMID- 10472503 TI - Kinship networks that cross racial lines: the exception or the rule? AB - I estimate the frequencies of interracial kin relations, an important indicator of the isolation of racial groups in the United States. I use two techniques to estimate the size and heterogeneity of extended families. First, I develop a simple model that takes account only of kinship network sizes and intermarriage levels by race. This model allows a crude estimation of the frequency of multiracial kinship networks. Second, I produce more precise empirical estimates using a new hot-deck imputation method for synthesizing kinship networks from household-level survey data (the June 1990 Current Population Survey and the 1994 General Social Survey). One in seven whites, one in three blacks, four in five Asians, and more than 19 in 20 American Indians are closely related to someone of a different racial group. Despite an intermarriage rate of about 1%, about 20% of Americans count someone from a different racial group among their kin. PMID- 10472504 TI - The leveling of divorce in the United States. AB - Is the recent plateau in crude divorce rates due to compositional changes in the married population or to a fundamental change in the long-term trend of rising marital instability? I use refined measures of period divorce rates to show that the leveling of divorce rates appears to be real. Compositional factors do little to explain the end to the more than century-long pattern of rising divorce. Increases in cohabitation also fail to explain the plateau. New theories are needed to explain the determinants of divorce rates at the population level. PMID- 10472505 TI - Trends in the intergenerational transmission of divorce. AB - I use data from the 1973-1996 NORC General Social Survey to examine trends in the intergenerational transmission of divorce, the propensity for the children of divorce to end their own marriages. The rate of divorce transmission declined by almost 50% in the study period. This result was essentially unchanged by statistical controls for various personal and family background differences between respondents. PMID- 10472506 TI - Chronic morphine reduces pain-related disability in a rodent model of chronic, inflammatory pain. AB - Chronic pain is disabling, and the adverse effects of morphine are also disabling. The best way to assess the beneficial effects relative to the potential adverse effects of chronic morphine may be through the use of quantitative measures of functional disability in people and animals experiencing pain. If chronic morphine alleviates chronic pain and its beneficial analgesic effects outweigh whatever adverse effects it may produce, then it should reduce pain-related disability. Rats with adjuvant-induced arthritis were implanted with subcutaneous morphine pellets. Continuous morphine reduced pain-related disability in tasks motivated by food reward or shock avoidance throughout the 35 days of continuous administration--first, in tests that primarily assessed the function of the less severely affected forelimbs, and later, as the inflammation subsided, in tests more dependent on the function of the more severely affected hind limbs. PMID- 10472507 TI - Regulation of intravenously self-administered nicotine in rats. AB - Ten male Wistar rats had access to 9 doses of nicotine (0.01-0.10 mg/kg i.v.) during daily 5-hr sessions. Once responding for nicotine stabilized, nicotine infusions were replaced with either cocaine infusions (0.0-2.4 mg/kg) or saline infusions. Saline substitution results indicate that nicotine functioned as a reinforcer. Regulation of nicotine intake was compared with that of cocaine by obtaining the correlation between mean interdose interval and preceding dose size. Results reveal that although this correlation was significant for both nicotine and cocaine self-administration, nicotine self-administration was less precisely regulated than cocaine self-administration. This procedure suggests that there are differences in regulation among self-administered drugs and that it may serve as a useful baseline for studying differences in vulnerability to drug abuse and potential treatment strategies. PMID- 10472508 TI - Effects of repeated methylphenidate treatment in the young rat: sensitization of both locomotor activity and stereotyped sniffing. AB - The behavioral effects of repeated methylphenidate (MPH) treatment were assessed in young rats. In 4 experiments, rats (starting at Postnatal Day 10 or 16) were pretreated on 5 consecutive days with saline or MPH (2.5-20.0 mg/kg i.p.). Sensitization was assessed after 1 or 7 abstinence days, with rats receiving a test day challenge injection of either a low dose of MPH (2.5 mg/kg) or the same dose of MPH as given during pretreatment. Results show that a test day injection of 2.5 mg/kg MPH produced a sensitized locomotor response in rats pretreated with 2.5-20.0 mg/kg MPH. This MPH-induced locomotor sensitization was evident only after 1 abstinence day. Various pretreatment doses of MPH (5, 10, 15, or 20 mg/kg) were capable of sensitizing the stereotyped sniffing of young rats, but only rats pretreated and tested with the highest dose (20 mg/kg) of MPH showed an augmented stereotyped sniffing response that was still robust after 7 abstinence days. Results indicate that young rats are capable of exhibiting sensitization after an extended abstinence period, which contrasts with previous research suggesting that psychostimulant treatment does not produce long-term sensitization in young rats. PMID- 10472509 TI - Comparison of morphine-induced effects on thermal nociception, mechanoreception, and hind limb flexion in chronic spinal rats. AB - The effect of systemic morphine on 3 behaviors in the same group of chronic spinal rats was examined: the tail-flick (TF) reflex to a noxious thermal stimulus, limb withdrawal (LW) to mechanoreceptor (von Frey hair) stimulation, and hind limb flexion (flexor reflex [FR]) elicited by innocuous electrical stimulation of the toes. Compared with intact rats, the potency of morphine on both the TF and the hind paw (but not the forepaw) LW response was significantly reduced. Morphine's effect on the FR depended on the dose. The lowest dose (1.0 mg/kg) produced no change, 4.0 mg/kg decreased response magnitude by approximately 50% (indicating an antispastic effect), and 8.0 mg/kg increased flexor magnitude by 100%. The concurrent TF and FR assays revealed a dissociation of morphine's effects in that the highest dose (8.0 mg/kg) significantly inhibited the nociceptive TF response but facilitated the FR in the same chronic spinal rats. This outcome may be relevant to the phenomenon of "opioid-related myoclonus" recently described in cancer patients, which "was highly associated with nerve dysfunction due to spinal cord lesions" (S. Mercadante, 1998, p. 6). PMID- 10472510 TI - Do heavy smokers benefit from higher dose nicotine patch therapy? AB - Heavy smokers (N = 408, smoking more than 25 cigarettes/day) were randomized to receive high-dose (25 mg) or standard-dose (15 mg) nicotine patch therapy delivered during the daytime only (16 hr) for a period of 6 weeks. Treatment effects of each dose were similar. The percentages of participants not smoking at 2-, 6-, and 12-month follow-ups were 26 versus 20, 14 versus 16, and 14 versus 14 for the 15-mg and 25-mg doses, respectively. The higher dose was well tolerated, and adverse event profiles for both treatment doses were very similar. Stepwise Cox proportional hazards analyses indicated that initial postrandomization craving and baseline scores on the Center for Epidemiological Studies Depression Instrument predicted time-to-relapse during treatment; only initial craving predicted relapse over the entire study interval (12 months). The results of this trial do not support the routine use of higher dose nicotine patch therapy in the treatment of nicotine dependence. PMID- 10472511 TI - Chronic tolerance and sensitization to alcohol in sons of alcoholics: II. Replication and reanalysis. AB - Data from D. B. Newlin and J. B. Thomson (1991) were reanalyzed, and data from an independent replication study were analyzed, relative to tonic (baseline) and phasic (response to alcohol challenge) aspects of drinking alcohol administered at the same dose on several occasions. Among the high-risk men (sons of alcoholic fathers), linear trends across days for resting (predrinking) baselines were opposite to alcohol-evoked changes for finger pulse amplitude, finger temperature, and skin conductance in Study 1 and for pulse transit time and body sway (static ataxia) in Study 2. In contrast, the structure of the low-risk men's (sons of nonalcoholic parents) data was precisely the opposite. Results are discussed in terms of sensitization as a potential mechanism that relates vulnerability to final manifestation of addictive behavior. PMID- 10472512 TI - Subjective effects of caffeine among introverts and extraverts in the morning and evening. AB - In previous studies of psychomotor performance, the stimulant effects of caffeine differed by personality characteristics. For example, caffeine improved the task performance of extraverts but overaroused introverts and thus impaired their performance. The present study compared the effects of caffeine on subjective arousal among introverts and extraverts. Seventeen introverts and 19 extraverts drank coffee that contained doses of 0, 2, and 4 mg/kg caffeine during morning and evening sessions in a within-subjects, randomized, double-blind, crossover design. At 30-min intervals for 180 min after drinking, participants completed the Profile of Mood States, a battery of self-report visual analog scales, and the Digit Symbol Substitution Test (DSST). Caffeine effects on mood and task performance did not significantly interact with extraversion, except for nonsignificant trends for caffeine to increase happiness and vigor more among extraverts than introverts. No 3-way interactions of group, time, and dose were found on any scales or on the DSST. Results do not support the hypothesis that caffeine differentially affects extraverts and introverts, particularly at different times of the day. PMID- 10472513 TI - Smoking cues decrease prepulse inhibition of the startle response and increase subjective craving in humans. AB - The present study investigated whether exposure to smoking cues would attenuate prepulse inhibition (PPI) of the startle reflex and increase craving among smokers across 2 experimental sessions. It was hypothesized that exposure to smoking cues would result in a decrease in PPI. Twenty-six smokers were exposed to smoking cues and control cues in 2 experimental sessions 1 week apart. Results indicate that smoking cues reliably attenuated PPI in both the 1st and 2nd sessions as compared with control cues. Findings also suggest that smoking cues reliably increased craving, increased negative affect, and reduced positive affect relative to baseline measures in both sessions. Results are consistent with the premise that exposure to smoking cues precipitates increases in dopamine activation or changes in information processing that cause a disruption of PPI. PMID- 10472514 TI - A follow-up study of the acute behavioral effects of benzodiazepine-receptor ligands in humans: comparison of quazepam and triazolam. AB - Quazepam, a trifluoroethylbenzodiazepine hypnotic, and triazolam, a triazolobenzodiazepine hypnotic, differ in terms of their benzodiazepine-receptor binding profile. Previous studies have suggested that quazepam produces less performance impairment than triazolam. Whether these effects are due to differences between quazepam and triazolam in terms of their benzodiazepine receptor binding profile or to the testing of insufficient doses is unknown. The present study compared the acute behavioral effects of triazolam (0.1875, 0.3750, and 0.5625 mg), quazepam (30, 60, and 90 mg), and placebo in 12 healthy humans using a within-subjects, placebo-controlled, crossover design. Quazepam and triazolam produced comparable dose-dependent performance impairment and increased ratings of drug effect and drowsy. Quazepam, but not triazolam, increased ratings of dizzy/light-headed, performance impaired, and sleepy. Triazolam, but not quazepam, increased ratings of high. Thus, across a sufficient range of doses, the performance-impairing effects of quazepam were similar to those of triazolam. By contrast, quazepam and triazolam produced somewhat different constellations of participant-rated drug effects. These differential drug effects may be attributable to differences between quazepam and triazolam in terms of their benzodiazepine-receptor binding profile. PMID- 10472515 TI - Influence of naltrexone on cue-elicited craving among hazardous drinkers: the moderational role of positive outcome expectancies. AB - This study was designed to elucidate mechanisms by which naltrexone (NTX) influences drinking among hazardous drinkers. Thirty-six hazardous drinkers received 50 mg NTX or placebo on 2 separate occasions before participation in a taste test procedure with low-alcohol beer. Urges to drink before consumption, beer volume consumed, and perceived stimulation and sedation after consumption were assessed. Although NTX did not influence beer consumption, hazardous drinkers who reported high positive reinforcement expectancies rated their urges to drink as significantly lower when they were on NTX compared with placebo. Positive outcome expectancies also moderated the effects of NTX on subjective reports of stimulation following drinking. These findings suggest that naltrexone may be particularly effective at reducing alcohol cue-elicited positive reinforcement for those with high positive alcohol outcome expectancies. PMID- 10472516 TI - Sex and menstrual cycle differences in the subjective effects from smoked cocaine in humans. AB - To investigate sex and menstrual cycle effects in response to cocaine administration, data from existing studies were analyzed. First, responses to a single delivery of 0.4 mg/kg smoked cocaine were investigated. Women reported lower ratings for measures of paranoid/suspicious and heart racing/pounding than did men. In addition, women in the luteal phase reported diminished ratings for a measure of feel high than did both women in the follicular phase of the menstrual cycle and men. Second, responses to up to 6 deliveries of 0.4 mg/kg smoked cocaine were investigated. Women, compared with men, had lower ratings on feel high, heart racing/pounding, and feel stimulated. Results suggest that there are significant sex and menstrual phase differences in the subjective effects of cocaine. PMID- 10472517 TI - Discounting of delayed rewards in opioid-dependent outpatients: exponential or hyperbolic discounting functions? AB - Recent theories of substance abuse have used value discounting of delayed rewards to partly explain the decision to take drugs. Normative-economic theory holds that an exponential function describes the effects of delay on discounting, whereas the matching law posits a hyperbolic discounting function. The ability of these functions to describe 18 human heroin-dependent individuals' monetary- and heroin-reward delay-discounting functions was assessed. In the 1st condition, participants chose between immediate and delayed hypothetical monetary rewards. Delayed rewards were $1,000, and the immediate reward amount was adjusted until choices reflected indifference. In the 2nd condition, participants chose between immediate and delayed heroin (the delayed amount was that which each participant reported he or she could purchase with $1,000). The hyperbolic function produced significantly higher R2 values and significantly lower sums of squared error values. Consistent with previous findings, delayed heroin rewards were discounted at a significantly higher rate than were delayed monetary rewards. PMID- 10472518 TI - Hypothesized simple factor structure for the Alcohol Expectancy Questionnaire: confirmatory factor analysis. AB - This study proposed and tested a theoretical factor structure for the Alcohol Expectancy Questionnaire (AEQ; S. A. Brown, M. S. Goldman, A. Inn, & L. R. Anderson, 1980). Factors were hypothesized to (a) reflect either positive or negative reinforcement, and (b) target either personal feelings or social contexts, resulting in 4 hypothesized factors (Social Enhancement, Social Coping, Personal Enhancement, Personal Coping). Participants were 180 male and 226 female undergraduates who completed the AEQ and additional self-report measures. Confirmatory factor analysis revealed that the hypothesized model fit the data. Post hoc adjustments further improved the model. Finally, a higher order factor model fit the data best. Factors correlated in hypothesized ways with other measures: (a) Only Personal Coping expectancies correlated with negative affect; (b) self-efficacy to resist drinking for emotional relief correlated highest with Personal Coping; and (c) self-efficacy to resist social pressure to drink correlated highest with Social Enhancement. Correlations with B. C. Leigh and A. W. Stacy's (1993) Drinking Expectancy Questionnaire indicated congruent and divergent validity. PMID- 10472519 TI - Catheter-induced vasospasm in the right external iliac and femoral arteries during a cardiac diagnostic procedure. AB - Catheter-induced vasospasm of small caliber arteries, such as the coronary arteries, is frequently observed during cardiac catheterization, but obstruction of the large caliber arteries has not previously been reported. Here we present two cases in which femoral and external iliac arteries were totally obstructed due to spasm during diagnostic coronary angiography. PMID- 10472520 TI - Comparison of dobutamine stress echocardiography and exercise stress Thallium-201 SPECT for detection of myocardial ischemia after acute myocardial infarction treated with thrombolysis. AB - In order to compare the ability of dobutamine stress echocardiography (DSE) and exercise Thallium-201 SPECT to detect myocardial ischemia in patients with myocardial infarction (MI) treated with thrombolysis, 43 prospectively selected patients with MI treated with thrombolysis underwent within 1 month from MI DSE, stress-redistribution-reinjection Thallium-201 SPECT and coronary angiography. The echocardiographic and scintigraphic images were analyzed for the presence of myocardial ischemia using a 11-segment left ventricular model. DSE and exercise Thallium-201 SPECT detected myocardial ischemia in the infarct zone in 72% and 72% (31/43) of patients and ischemia at a distance in 12% (5/43) and 19% (8/43) of patients with a concordance of 67% and 88%, respectively. A significant agreement between DSE and exercise Thallium SPECT was found in the evaluation of the extent of both myocardial necrosis and stress-induced myocardial ischemia. DSE and exercise Thallium SPECT showed similar sensitivity (79 vs 76%), specificity (60 vs 60%) and accuracy (77 vs 74%) for detection of a critical stenosis of the infarct-related artery; there was also no significant difference between the tests in sensitivity, specificity and accuracy for detection of the multivessel disease. In conclusion, initially after thrombolyzed MI, DSE and exercise Thallium-201 SPECT detect myocardial ischemia in the infarct zone in a high proportion of patients and show a similar accuracy for the diagnosis of a critical stenosis of the infarct-related coronary artery and of the multivessel disease. PMID- 10472521 TI - Neovascularity related to mural thrombus in endomyocardial fibrosis. AB - We report a 30-year old, previously healthy, Ghanese woman admitted with sudden onset of dyspnoe, hemoptoe and right-sided chest pain due to endomyocardial fibrosis with secondary pulmonary emboli Coronary angiography revealed a myocardial "blush". This finding may focus attention to the presence of mural thrombus that may have diagnostic and therapeutic consequences. PMID- 10472522 TI - Dynamic obstruction, an unusual complication after aortic valve replacement with a stentless porcine valve. AB - In the early nineties, the stentless porcine aortic bioprosthesis has been reintroduced successfully. Because of the limited experience, knowledge of clinical complications is limited. Therefore, we describe an unusual complication of dynamic obstruction after aortic valve replacement with a stentless porcine valve in a 70 year old man 18 months after implantation. We discuss the complications of stentless aortic prostheses known so far, describe operative techniques used and their characteristic two dimensional echocardiographic images. PMID- 10472523 TI - Harmonic imaging: echocardiographic enhanced contrast intensity and duration. AB - The intensity and duration of contrast effect within the left ventricular cavity after an intravenous bolus of Levovist Injection were observed with both harmonic and fundamental imaging in nine patients with known or suspected coronary artery disease. Contrast intensity was assessed by a qualitative grading system (0, none; 1, weak; 2, moderate; 3, good) and by videodensitometric analysis of pixel intensity. Duration of left ventricular contrast effect was determined by measuring time from the initial visual appearance of contrast agent to its disappearance. The mean increase in pixel intensity within the left ventricular cavity from precontrast to peak contrast was significantly greater for second harmonic than for fundamental imaging (25.5 vs 7.1; P < 0.012). The mean contrast intensity qualitative score with harmonic imaging was higher (2.6 +/- 0.73 vs 1.2 +/- 0.44; P < 0.01) and the duration of contrast effect was longer (242 +/- 131 s vs 53 +/- 33 s; P < 0.004). Second harmonic imaging significantly enhanced contrast intensity and prolonged visible duration of contrast effect after a peripheral venous injection of Levovist. PMID- 10472524 TI - Comparison of normal and diseased pulmonary artery morphology by intravascular ultrasound and histological examination. AB - The study was performed to determine the morphological characteristics of normal and diseased pulmonary arteries by ultrasound (intravascular ultrasound, IVUS) and histology. Forty-nine cadaver segments of pulmonary arteries from 16 postmortem patients were imaged in vitro by IVUS and compared to matched histological sections. The pulmonary vasculature of 11 patients with pulmonary hypertension was investigated in vivo by IVUS. In the in vitro study, 34 of a total of 143 histological sections of the segmental pulmonary arteries showed fibrotic wall components; the remaining 109 sections had regular components. Imaged by IVUS, the wall of the regular and fibrotic arteries revealed a single layer histologically, representing the intima, media, adventitia and connective tissue. Adjacent lung tissue could be detected by IVUS and was confirmed by the histological section. In three patients with a history of chronic embolic pulmonary hypertension, IVUS revealed thrombi or a double vessel wall layer. Histologically, the material extracted after pulmonary thromboendarterectomy in two patients showed wall-adherent thrombus features of early organization. In all other patients only a single wall layer was seen. The vessel walls (intima, media, adventitia and connective tissue) of regular and slightly fibrosed pulmonary arteries show only a single layer. Wall-adherent organized thrombi in chronic embolic pulmonary hypertension can be detected by IVUS as a second inner vessel layer. Thus IVUS may represent an additional tool for detecting chronic embolic pulmonary hypertension when the results of pulmonary angiography or computed tomography are not definite. PMID- 10472525 TI - Worsening of left ventricular diastolic function during long-term correction of anemia with erythropoietin in chronic hemodialysis patients--an assessment by radionuclide ventriculography at rest and exercise. AB - We studied the effect of correction of anemia with erythropoietin on left ventricular systolic and diastolic function at rest and exercise in 17 chronic hemodialysis patients by means of maximum exercise testing and equilibrium gated radionuclide angiocardiography on three occasions: 1) initial--before erythropoietin administration, 2) intermediate--at the time when the target hemoglobin level reached 100 g/l, and 3) long-term--after 12 months of therapy. After correction of anemia, the patients showed a significant improvement in their response to exercise regarding maximal work load achieved, exercise duration and recovery time. Ejection fraction and peak ejection rate remained unchanged during therapy. At rest, peak filling rate was reduced from 2.62 +/- 1.0 (baseline) to 2.28 +/- 0.9 (intermediate) end-diastolic volume per second, p < 0.01, while no significant difference was observed during exercise. The time to peak filling rate was prolonged significantly during EPO therapy from 157 +/- 30 to 177 +/- 28 ms at rest, p < 0.05, and from 101 +/- 24 to 130 +/- 27 ms during exercise, p < 0.01. By the time of the late study, there were no significant differences between the late and intermediate study. In conclusion, amelioration of anemia with erythropoietin in hemodialysis patients produced improvement in exercise capacity, but diastolic function worsened with therapy and this effect was maintained during the long-term treatment, while systolic function at rest and exercise remained unchanged. PMID- 10472526 TI - Kinematic analysis of left ventricular deformation in myocardial infarction using magnetic resonance cardiac tagging. AB - The Magnetic Resonance (MR) tagging technique provides detailed information about 2D motion in the plane of observation. Interpretation of this information as a reflection of the 3D motion of the entire cardiac wall is a major problem. In finite element models of the mechanics of the infarcted heart, an infarcted region causes motional asymmetry, extending far beyond the infarct boundary. Here we present a method to quantify such asymmetry in amplitude and orientation. For this purpose images of a short-axis cross-section of the ejecting left ventricle were acquired from 9 healthy volunteers and 5 patients with myocardial infarction. MR-tags were applied in a 5 mm grid at end-diastole. The tags were tracked by video-image analysis. Tag motion was fitted to a kinematic model of cardiac motion. For the volunteers and the patients the center of the cavity displaced by about the same amount (p = 0.11) during the ejection phase: 3.8 +/- 1.4 and 3.0 +/- 0.9 mm (mean +/- sd), respectively. Cross-sectional rotation and the decrease in cross-sectional area of the cavity were both greater in the volunteers than in the patients: 6.4 +/- 1.5 vs. 3.0 +/- 0.8 degrees (p < 0.001), and 945 +/- 71 vs. 700 +/- 176 mm2 (p = 0.02), respectively. In the patients, asymmetry of wall motion, as expressed by a sine wave dependency of contraction around the circumference, was significantly enlarged (p = 0.02). The proposed method of kinematic analysis can be used to assess cardiac deformation in humans. We expect that by analyzing images of more cross-sections simultaneously, the 3D location and the degree of infarction can be assessed efficiently. PMID- 10472527 TI - Diagnosis and management of anomalous origin of the right coronary artery from the left coronary sinus. AB - The anomalous origin of the right coronary artery (RCA) from the left sinus of Valsalva coursing between the aorta and the pulmonary artery or right ventricular outflow tract, is considered a potentially fatal abnormality which may require surgery. However, diagnosing the correct course with coronary arteriography may be difficult. Fast gradient echo magnetic resonance (MR) imaging can be helpful to identify and confirm the course of aberrant coronary arteries and their relationship to the surrounding tissue. In this study, diagnostic procedures and management are described of four patients in whom the RCA originated from the left sinus of Valsalva. Although reported as investigational by the Task Force document on MR imaging by the European Society of Cardiology we are of the opinion that MR coronary angiography may have an important future role in the assessment of anomalous coronary arteries. PMID- 10472528 TI - Diagnostic uses of DNA analysis: powerful but problematic. PMID- 10472529 TI - Large heterozygous deletion masquerading as homozygous missense mutation: a pitfall in diagnostic mutation analysis. AB - The clinical use of molecular analyses in recessive disorders relies on the exact characterization of both mutant alleles in the affected patient. This can be problematic when only part of the gene is examined or when relevant DNA alterations are not recognized by standard methods. We present a child in whom phenylketonuria was apparently caused by homozygosity for the mutation E390G in exon 11 of the phenylalanine hydroxylase (PAH) gene. However, the clinical severity of the disease was not quite as mild as expected, the mutation was not identified in the father despite confirmed paternity, and the paternal allele showed a highly unusual pattern of polymorphic markers in the PAH gene. Presence of a large deletion involving exons 9, 10 and 11 of the phenylalanine hydroxylase gene was confirmed by long-range PCR. Diagnostic DNA analyses should include a comprehensive examination of the whole relevant gene in the patient and confirmation of carrier status in both parents. PMID- 10472530 TI - Regression of neuropsychological deficits in early-treated phenylketonurics during adolescence. AB - Even early-treated phenylketonuric patients suffer from phenylalanine-associated (mild) neuropsychological impairment. To date it is still unclear whether patients' deficits show a progression on ageing. This unsolved question seems to be an important aspect in the still ongoing debate about how long and how strictly the patients should be maintained on diet. Twenty early-treated (20 +/- 10, 9-30 days) adolescent phenylketonurics (10 boys, 10 girls) and 20 healthy controls, matched for age, sex and IQ, were investigated twice at a mean ages of 11 and 14 years for their IQ (Culture Fair Intelligence Test-Scale 2; CFT-20), fine motor abilities (Motor Performance Task), sustained (Test d2) and selective attention (Stroop-Task). At the first test, examinations revealed significant blood phenylalanine-correlated neuropsychological deficits in PKU patients. In spite of raised blood phenylalanine concentrations during the following 3 years and significantly elevated concurrent blood phenylalanine concentrations, the repeated measurements revealed a significant decrease of patients' deficits compared to controls. Clinical-neurological status of patients and controls was normal at both test times. The results indicate a decreased vulnerability of PKU patients with respect to their neuropsychological functioning against elevated blood phenylalanine levels on ageing. PMID- 10472531 TI - Renal clearance of branched-chain L-amino and 2-oxo acids in maple syrup urine disease. AB - In maple syrup urine disease (MSUD), branched-chain L-amino (BCAA) and 2-oxo acids (BCOA) accumulate in body fluids owing to an inherited deficiency of branched-chain 2-oxo acid dehydrogenase complex activity. In MSUD, little information is available on the significance of urinary disposal of branched chain compounds. We examined the renal clearance of leucine, valine, isoleucine and alloisoleucine, and their corresponding 2-oxo acids 4-methyl-2-oxopentanoate (KIC), 3-methyl-2-oxobutanoate (KIV), (S)-(S-KMV), and (R)-3-methyl-2 oxopentanoate (R-KMV), using pairs of plasma and urine samples (n = 63) from 10 patients with classical MSUD. The fractional renal excretion of free BCAA was in the normal range (< 0.5%) and independent of the plasma concentrations. The excretion of bound (N-acylated) BCAA was normal and not significantly dependent on the BCAA plasma concentrations. The fractional renal excretion of BCOA was in the order KIC << KIV < R-KMV < or = S-KMV (range (%): KIC 0.1-25; KIV 0.14-21.3; S-KMV 0.26-24.6; R-KMV 0.1-35.9), significantly correlated with the KIC plasma concentrations, and generally higher than that of the related BCAA. The results show that the renal excretion of free BCAA as well as of the acylated derivatives is negligible. The renal excretion of BCOA, however, to some extent counteracts increases in BCAA concentrations and thus contributes to the lowering of total BCAA pools in MSUD. PMID- 10472532 TI - Long-term outcome of liver transplantation in patients with glycogen storage disease type Ia. AB - Liver transplantation may be indicated in patients with GSD type Ia when dietary treatment fails or when hepatic adenomas develop, because they carry a risk of liver cancer or severe intratumoral haemorrhage. Published reports on the results of liver transplantation in patients with GSD Ia include 10 patients and provide little information on long-term outcome. In particular, it is not known whether liver transplantation prevents renal failure due to focal segmental glomerulosclerosis. We report here on 3 patients with GSD Ia in whom liver transplantation was performed at 15, 17 and 23 years of age because of multiple hepatic adenomas in all 3 patients with a fear of malignant transformation, and of poor metabolic balance and severe growth retardation in the youngest one. Renal function was normal in all patients. During the 6-8 years following transplantation, the quality of life has initially greatly improved, with none of the previous dietary restraints and a spectacular increase in height. However, long-term complications included chronic hepatitis C in one patient, gouty attacks in another and focal segmental glomerulosclerosis with progressive renal insufficiency in the third. These results: (1) confirm that liver transplantation restores a normal metabolic balance in patients with GSD Ia, allows catch-up growth and improves the quality of life; (2) suggest that liver transplantation may be considered in teenagers with unresectable multiple adenomas because of a lack of clear-cut criteria to detect malignant transformation early; and (3) suggest that liver transplantation does not prevent focal segmental glomerulosclerosis associated with GSD Ia. PMID- 10472533 TI - Medium-chain triglyceride loading test in carnitine-acylcarnitine translocase deficiency: insights on treatment. AB - The results of a medium-chain triglyceride loading test in a patient with severe carnitine-acylcarnitine translocase deficiency clearly demonstrated impaired in vivo utilization of medium-chain triglycerides. The loading test was performed at the ages of 7 and 36 months. The diet was adjusted accordingly. The clinical course has been favourable and the child is now in very good condition at age 4 years. We conclude that the utilization of medium-chain triglycerides is only partial in carnitine-acylcarnitine translocase deficiency and cannot reasonably be considered an optimal source of energy for these patients. Careful adjustment of dietetic treatment may help to improve prognosis. PMID- 10472534 TI - A simple screening test for fatty acid oxidation defects using whole-blood palmitate oxidation. AB - We report that measurement of whole-blood palmitate oxidation is a rapid and inexpensive screening test for fatty acid oxidation defects. The assay has been adapted from published assays using cultured fibroblasts or isolated white blood cells. Micro whole-blood samples are incubated with tritiated palmitic acid as substrate. The tritiated water produced is proportional to the mitochondrial beta oxidation of palmitic acid. Patients with confirmed beta-oxidation defects show low whole-blood palmitate oxidation. PMID- 10472535 TI - Intermittent jaundice in patients with acute leukaemia: a common mutation of the bilirubin uridine-diphosphate glucuronosyltransferase gene among Asians. AB - The Gly71Arg mutation of the hepatic bilirubin UDP glucuronosyl-transferase (B UGT) gene associated with Gilbert syndrome prevails among Japanese and its gene frequency is 0.13. Among 20 patients with acute leukaemia, 4 patients showed intermittent unconjugated hyperbilirubinaemia during the course of combined chemotherapy. The Gly71Arg mutation was detected in all 4 patients with hyperbilirubinaemia, but was not found in 16 patients without hyperbilirubinaemia. Two of them were heterozygotes and one was a homozygote for the Gly71Arg mutation, and the other was a compound heterozygote of the Gly71Arg mutation and TA insertion mutation in the TATA box of the B-UGT gene. In addition to the complications leading to hyperbilirubinaemia, including liver damage due to drugs, viral infections or tumour cell infiltrations and alloimmune haemolysis, carrier status for the Gly71Arg mutation should be considered in a patient with leukaemia showing intermittent hyperbilirubinaemia during the course of chemotherapy, especially among Japanese, Koreans and Chinese owing to its prevalence in those populations. PMID- 10472536 TI - Spontaneous pregnancy in a patient with classical galactosaemia. PMID- 10472537 TI - Reversible dementia in an adolescent with cblC disease: clinical heterogeneity within the same family. PMID- 10472538 TI - Generalized glutathione synthetase deficiency and pregnancy. PMID- 10472539 TI - The Leber hereditary optic neuropathy 14,484 mutation and X-linked adrenoleukodystrophy: a possible modifier of phenotypic expression? PMID- 10472540 TI - Novel donor splice site mutations of AGL gene in glycogen storage disease type IIIa. PMID- 10472541 TI - Parent behavior and child distress during urethral catheterization. AB - ISSUES AND PURPOSE: Researchers need a clear understanding of the natural behaviors parents use to help their children cope. This study describes the relationships between naturally occurring parent behaviors and child distress behaviors during urethral catheterization. DESIGN AND METHODS: In this descriptive study, researchers videotaped the behaviors of parent-child interactions during urethral catheterization. RESULTS: Parents used distraction to maintain calm behavior during the first part of the procedure and used more reassurance when the children started to become distressed. Seven of the nine children displayed calm behavior at least half the time following distraction. Parental reassurance did not decrease distress behavior in most children. PRACTICE IMPLICATIONS: Early implementation of developmentally appropriate nursing interventions to decrease child distress is imperative. Parents may need specific instruction and practice to continue the use of distraction throughout procedures, even when the child is upset. PMID- 10472542 TI - Pediatric registered nurse usage and perception of EMLA. AB - ISSUES AND PURPOSE: Pediatric nurses do not consistently use the anesthetic cream for children. This study examined availability, use, and perception of driving and restraining forces for use of EMLA cream for venipuncture and i.v. catheter insertion. DESIGN AND METHODS: In this descriptive study, surveys were sent to 400 randomly selected pediatric RNs. Surveys were returned by 211 nurses from 36 states. RESULTS: Driving forces for EMLA usage included ease of use and increased patient rapport. Restraining forces were focused on barriers to accessing EMLA. Positive perceptions of EMLA were affected by ease of accessibility. PRACTICE IMPLICATIONS: Use of EMLA may be enhanced by standard orders, inservice instruction, and stocking EMLA on the patient care unit. PMID- 10472543 TI - The misunderstood child: the child with a nonverbal learning disorder. AB - ISSUES AND PURPOSE: To introduce nurses to the characteristics of nonverbal learning disorders (NLD) and their psychosocial trajectory in children. Despite the prevalence of learning disorders in children, few nursing professionals are familiar with NLD and their manifestations. CONCLUSIONS: The child with NLD faces unique challenges in the social, academic, visual-spatial, motoric, and emotional spheres. PRACTICE IMPLICATIONS: Nurses can play a vital role in the identification, education, and provision of resources for these children. PMID- 10472544 TI - Listening to Hispanic mothers: guidelines for teaching. AB - Teaching Hispanic mothers in a culturally sensitive way to care for their children is a challenge to pediatric nurses. Pediatric nurses must be familiar with customs and the folk medicine practiced by Hispanic mothers. It is very important that the pediatric nurse listens to the voices of Hispanic mothers to determine their health practices, and those that may have been used in their children. Familiarity with folk medicine and health practices will facilitate an appropriate treatment plan and will help to determine whether the mothers' practices are dangerous or beneficial for the child. Pediatric nurses should assess for concurrent use of home remedies and conventional medications to determine if there are any known interactive effects. Finally, increasing the number of pediatric nurses who are fluent in Spanish will enable the voices of Hispanic mothers to be better heard, which in turn, will improve the health status of Hispanic children. PMID- 10472545 TI - Identifying and caring for the child with new onset type 1 diabetes. AB - The parents of a child with new onset type 1 diabetes must master essential "survival skills" to return home safely with their child. It is the healthcare provider's responsibility to assure that the family has been taught necessary information to succeed at home and provide the family with access to nurses and physicians for insulin adjustment and emergencies upon discharge. PMID- 10472546 TI - [Expert network for pediatric oncology and hematology]. PMID- 10472547 TI - [Computer-assisted documentation and therapy planning in pediatric oncology- introduction of a nationwide solution]. AB - In Pediatric Oncology in Germany, 90% of the patients are treated according to multicenter clinical trials, which means an enormous effort for documentation in the participating clinics. In order to enable multiple use of data for patient records as well as for clinical trials a computer-based documentation system for pediatric oncology (DOSPO) is being developed, which can be used nationwide. DOSPO currently comprises a minimum basic data set, which represents the common core of all multicenter trials and which has been approved by the German Society for Pediatric Oncology and Hematology (GPOH). It is intended to enhance the documentation by specific items of each clinical trial. Functions for computer aided chemotherapy planning and medical report writing have already been implemented in the documentation system. Various centers in Germany are currently validating the system in routine use. PMID- 10472548 TI - [Molecular basis of Fanconi's anemia]. AB - Fanconi anaemia (FA) is an autosomal recessive genetic disorder characterised clinically by progressive bone marrow failure, skeletal deformities and a predisposition to neoplasia. Patient cells manifest an extreme chromosomal instability and hypersensitivity to polyfunctional alkylating agents. It is assumed that the basic defect is related to the repair of DNA damage, in particular that of so-called DNA crosslinks. Currently there are eight complementation groups in FA (FA-A-FA-H) which indicates that at least eight independent genes can lead to FA. Three of these genes have been identified: FANCA, FANCC and FANCG. In this review, the molecular biology and genetics of FA are presented and possible functions of the FANC proteins are discussed. PMID- 10472549 TI - [Thrombocytic alpha-delta-storage-pool-disease: shortening of bleeding time after infusion of 1-desamino-8-D-arginine vasopressin]. AB - BACKGROUND: The synthetic vasopressin derivate desmopressin (1-desamino-8-D arginine vasopressin) has been reported to shorten the bleeding time in patients with hemophilia A, von Willebrand's disease and several functional platelet disorders. In addition to substitution of platelets, vasopressin is therefore used to prevent bleeding complications. CASE: We report the case of a 14-year-old female patient with prolonged bleeding time due to the rare thrombocytic alpha delta-storage-pool-disease. When normal donor platelet substitution alone was ineffective, bleeding time was normalised after infusion of desmopressin and elective wisdom-tooth extraction was performed without significant postoperative bleeding. DISCUSSION: Infusion of desmopressin appears to be effective in shortening bleeding time in thrombocytic storage-pool-disease. Its use could prevent bleeding complications after trauma and surgical interventions and may possibly help to spare the need for platelet concentrates. PMID- 10472550 TI - Bleeding and thrombosis in children with acute lymphoblastic leukaemia, treated according to the ALL-BFM-90 protocol. AB - A multi-center retrospective survey was conducted to evaluate the incidence and types of hemostatic complications occurring in children with acute lymphoblastic leukemia (ALL) during treatment according to the ALL-BFM-90 treatment protocol. All of the BFM-treatment centers (n = 77) were approached and a 95% response rate with information on 1100 patients was obtained. Thrombotic or bleeding episodes occurred in 31 patients (2.8%), 19 of whom had thrombosis and 12 bleeding complications, involving the central nervous system (42%), the subclavian vein (29%), the gastro-intestinal tract, skin, lower extremities or pelvis (29%). Recovery was noted in 28 of 31 patients, while 3 died as a result of hemostatic complications. Bleeding or thrombosis occurred in patients receiving prophylactic substitution with plasma or plasma-derived concentrates (n = 16) as well as in those without substitution (n = 13). The majority of hemostatic complications (90%) occurred during the induction therapy of the treatment protocol, in particular during the period which included simultaneous administration of glucocorticoids and E. coli L-asparaginase. The concurrent administration of E. coli L-asparaginase and glucocorticoids may be an additional risk factor for thromboembolic events during therapy according to the ALL-BFM-90 protocol. PMID- 10472551 TI - Effect of glucocorticoids, E. coli- and Erwinia L-asparaginase on hemostatic proteins in children with acute lymphoblastic leukemia. AB - BACKGROUND: The concentrations of plasma hemostatic proteins were analyzed prospectively in 42 children with acute lymphoblastic leukemia (ALL), treated according to the protocol ALL-BFM-90. PROCEDURE: Treatment included glucocorticosteroids (GC), E. coli L-asparaginase (Asparaginase, Medac) or Erwinia L-asparaginase (Erwinase, Speywood), vincristine, anthracyclines and intrathecal methotrexate. The analysis of hemostatic proteins was performed during induction and re-induction therapy. RESULTS: At diagnosis, the plasma concentrations of fibrinogen, antithrombin III (AT), plasminogen and protein C were within normal limits, whereas the von Willebrand factor antigen (vWF:Ag) was elevated. After eight days of mono-therapy with GC the concentration of fibrinogen decreased to 59%, vWF:Ag decreased to 67%, AT increased to 124%, protein C increased to 201% of the initial value (mean all p < or = 0.01), while the concentration of plasminogen remained unchanged. During the re-induction phase, the concentrations of the hemostatic proteins, with exception of vWF:Ag, altered in a similar way in response to GC as observed during the induction phase. Administration of two doses of E. coli L-asparaginase (10,000 U/m2) during the induction therapy led to a significant decrease of AT (123 +/- 24 to 63 +/- 15%/mL), protein C (168 +/- 34 to 87 +/- 19%/mL), plasminogen (94 +/- 21 to 41 +/ 12%/mL) and fibrinogen (148 +/- 59 to 79 +/- 30 mg/dL, p < or = 0.01 for all parameters). In contrast, administration of two doses of Erwinia L-asparaginase (10,000 U/m2) during re-induction therapy did not lead to change in the concentration of AT, protein C or plasminogen, and the decrease in fibrinogen (162 +/- 17 to 121 +/- 24 mg/dL) was less pronounced. CONCLUSIONS: Our results indicate that GC and E. coli L-asparaginase, in particular, induce hemostatic alterations which have implications on our understanding of thrombotic and hemorrhagic events during the treatment of ALL in children. PMID- 10472552 TI - Cerebral venous sinus thrombosis in children with acute lymphoblastic leukemia carrying the MTHFR TT677 genotype and further prothrombotic risk factors. AB - BACKGROUND: The present study was designed to prospectively evaluate the role of prothrombotic risk factors in leukemic children treated according to the ALL-BFM 90/95 study protocols with respect to the onset of cerebral venous sinus thrombosis. PATIENTS: 317 consecutive leukemic children aged 6 months to 18 years were enrolled in this study. 288 of the 317 patients were available for thrombosis-free survival analysis. RESULTS: In 17 (5.9%) of these 288 patients cerebral venous sinus thrombosis occurred. The overall event-free survival of thrombosis in the central nervous system in patients with at least one prothrombotic defect (n = 12) was significantly reduced compared with patients without a prothrombotic defect (p < 0.0001). 15 patients showed acute clinical symptoms at onset of cerebral venous sinus thrombosis, two were asymptomatic. Three of the 17 patients affected (17.6%) died directly associated with the thrombotic event during induction therapy, the remaining 14 patients did not show prolonged clinical symptoms. CONCLUSIONS: Prothrombotic risk factors should be included in a screening program in ALL children treated according to the BFM study protocols. Further prospective studies are recommended to establish adequate prophylactic anticoagulant treatment during ALL (BFM) polychemotherapy. PMID- 10472553 TI - Low rate of severe venous thromboses in children with ALL treatment according to COALL-92 and -97 protocol. AB - Venous thromboses (VT) in children with ALL who were treated according to the COALL-89 protocol were reported to occur with a frequency of 2.1% (6/286). 4/6 of the reported VT were catheter related. However, in other cohorts of ALL patients treated according to American protocols the incidence of severe thromboses was 2 11%. Most of the VT were not catheter related, but were atypical thromboses like sinus venous thromboses. In these patients hereditary thrombophilia risk factors seemed to play a major role. In a 6 year period including the COALL protocols -92 and -97 only 10/684 (1.5%) children presented with symptomatic VT, and 7/10 thromboses were catheter related. Every thrombotic event could be successfully treated either by heparin administration or fibrinolytic agents. 2/10 VT were secondary due to a septic event. 5 out of 8 primary VT occurred after asparaginase/dexamethasone application during the reinduction therapy. In conclusion, symptomatic thrombotic events are very rare in the COALL studies. Important risk factors for development of VT appeared to be central lines, asparaginase application and infectious/septic complications. However, the role of genetic risk factors of thrombosis in these patients has still to be determined. PMID- 10472554 TI - Donor selection process for allogeneic hematopoietic stem cell transplantation at the university hospital of Dusseldorf (1997-1998). AB - BACKGROUND: Transplantation of hematopoietic stem cells (HSC) is an effective treatment for a number of patients with life-threatening hematologic diseases. HSC donors can be found in the family of the patient or in registries of unrelated donors. In the present study, the search procedure within the last two years for an allogeneic HSC donor at the University of Dusseldorf is analyzed. PATIENTS AND METHODS: During 1997 and 1998, an early search for a related HSC donor in the family was performed for 70 high risk pediatric patients. During the same period, the search for an unrelated HSC donor for 116 adult and pediatric patients was performed. Low resolution HLA-A and -B typing was performed by serology in combination with DNA-typing. High resolution typing of HLA-A, -B and C was carried out by DNA-sequencing. Low resolution HLA-DRB- und HLA-DQB1-typing was done solely by DNA-typing and high resolution typing of these genes was performed by DNA-sequencing. MAIN RESULTS: For 51 of 70 high risk pediatric patients (73%), no family donor could be defined, 16 of 70 patients (23%) had a genotypically identical sibling and for three of 70 patients (4%) an HLA acceptable donor in the extended family could be identified. The search for an unrelated HSC donor was successful in 74% of the adult and pediatric patients lacking such a family donor. Most noteworthy, nearly all of the HLA-acceptable donors were identified from that group of donors in the registries, which were not only HLA-A and HLA-B, but also HLA-DR pretyped. CONCLUSION: These data show, that a growing number of pediatric patients with high risk leukemia need an unrelated HSC donor and that HLA-ABDR-pretyped registries present the optimal prerequisite to identify an HSC donor for most of the patients. In addition, 25% of the patients with no family or unrelated HSC donor require HSC transplants from alternative donors like unrelated Cord Blood (CB) from high quality cord blood banks. PMID- 10472555 TI - Haematopoietic transplant potential of unrelated and related cord blood: the first six years of the EUROCORD/NETCORD Bank Germany. AB - To date, human umbilical cord blood (CB) has been employed successfully in well over 1000 allogeneic (unrelated and sibling) stem cell transplantations. Because of primary limitations in volume and cell numbers, over 90% of these transplantations were performed in children. Therefore requests for well standardised cord blood units of high quality are now increasing constantly. Examination and standardisation of unrelated and related cord blood stem cell preparations and banking as well as their biological characterisation was already initiated in Dusseldorf in 1992. Hitherto a total of 3236 CB samples with a mean volume of 89 +/- 25 ml, a mean total number of nucleated cells (NC) of 10 +/- 5 x 10(8) and a mean number of CFU-GM of 6 +/- 5 x 10(5) have also been validated by haematological, immunological and microbiological criteria. In addition to that, 97 directed CB donations of siblings with a clinical indication have been characterised and banked along the same lines. All CB units were collected from the umbilical cord vein immediately after vaginal full term delivery or caesarean section, then frozen and stored in liquid nitrogen. 1940 CB units were stored unseparated, the other 1296 were volume reduced using Hetastarch (HES) with a mean recovery of 85 +/- 13% of the nucleated cells, 86 +/- 12% and 84 +/- 13% for CFC and CD34+ cells, respectively. Only 5.0 ml of a CB sample is required for routine laboratory testing as there are HLA-class I typing, HLA-class II typing by sequence specific oligonucleotide probes (PCR-SOP), ABO typing, sterility control, assessment of progenitor and stem cells by colony forming assays, and CD34+ status as well as certain viral infections such as CMV, Hepatitis B, C, HIV, Parvo B19 by PCR technology before releasing the CB unit for transplantation. For apparent viral infections, maternal sera obtained at birth were tested for HBsAg, anti-HBc, anti-HCV, -HAV-(IgG, IgM), -HIV-1-2, -EBV- (IgG, IgM), -HTLVI-II, -CMV (IgM, IgG), toxoplasmosis and syphilis. Within the last three years a total of 4860 preliminary searches and 680 extended unit reports were submitted to the CB bank Dusseldorf by fax or World Wide Web. So far 68 unrelated and 3 related CB units were delivered. From these 70 have been transplanted in 30 different transplant centres world-wide. Until now the evaluation of the first 53 unrelated CB-transplantations was performed together with the EUROCORD transplant registry. Three patients were excluded from the analysis, since they received an unrelated CB-transplant for non-engraftment after previous allotransplants. The median patient age of these 50 patients was 5.0 years (range 0.3-44), the median weight 18 kg (range 4-70 kg). The majority of the patients transplanted for malignancies (66%) suffered from ALL (n = 19), AML (n = 7), CML (n = 4) and lymphoma (n = 2) with two third (75%) in an intermediate (2nd CR) or advanced stage of disease (> 2nd CR); 13 patients had metabolic diseases and immunodeficiencies and three had aplastic anaemia. All CB samples as well as the patients' blood samples were typed in Dusseldorf for HLA class I by serology confirmed by PCR-SSP and by high resolution DNA typing for HLA-DRB1 and HLA-DQB1 alleles. 96% of the 50 patients receiving unrelated CB were mismatched at one or more HLA-antigens. 41 of the 50 patients transplanted with unrelated CB from Dusseldorf were evaluable for engraftment with an overall engraftment rate of 83%. According to the defined criteria of EUROCORD, 9 of the 50 patients were not evaluable for engraftment, since they died before day 60. The present median follow-up time is 14 months (1.4-38). The Kaplan-Meier estimate of survival at one year is 42 percent. The three paediatric patients after sibling CB transplantations (ALL, amegakaryocytic thrombocytopenia and CML) are alive with a follow-up time of 350, 379 days and 531 days. (ABSTRACT TRUNCATED) PMID- 10472556 TI - [In vitro drug resistance as independent prognostic factor in the study COALL-O5 92 Treatment of childhood acute lymphoblastic leukemia; two-tiered classification of treatments based on accepted risk criteria and drug sensitivity profiles in study COALL-06-97]. AB - BACKGROUND: Risk-adapted therapy in acute lymphoblastic leukemia (ALL) of childhood relies on traditional risk factors such as age, white blood count, immunological subtype, chromosomal aberrations and response to treatment. In spite of risk-adapted therapy, however, 20-30% of the patients suffer a relapse and may have profited from more intensive therapy. On the other hand a third of the patients is probably overtreated considering cure rates of 30% with less intensive therapies in the seventies. Additional prognostic criteria are therefore urgently needed. In a retrospective Dutch study in childhood ALL drug sensitivity testing with the MTT assay was identified as a new highly significant prognostic factor. PATIENTS AND METHODS: In study COALL-92 in 140 patients in vitro drug sensitivity was performed on initial bone marrow or blood samples and as in the Dutch study a score was derived from the sensitivity of the 3 drugs prednisolone, vincristine and asparaginase (PVA score). For each drug a score of 1 (highest) to 3 (lowest sensitivity) is given. A score of 3 therefore indicates the best, a score of 9 the worst sensitivity. RESULTS: Probability of event-free survival (pEFS) according to the PVA score was 0.94 for score 3 + 4, 0.80 for score 5-7, 0.35 for Score 8 + 9 (0.47 for score 7-9). For analysis within the low risk (LR) group (age 1-10 years, WBC < 25/nl, common or pre-B-ALL, remission day 28, no translocation 9;22 or 4;11) and high risk (HR) group 252 patients of the Dutch study and the COALL study were combined. In the LR group pEFS was 1.00 for score 3 + 4, 0.76 for score 5-7, 0.38 for score 8 + 9 (0.54 for score 7-9); in the HR group pEFS was 0.91 for score 3 + 4, 0.69 for score 5-7, 0.45 for score 8 + 9 (0.56 for score 7-9). Multivariate analysis identified the PVA score as independent prognostic factor. NEW STUDY COALL-97: In the new study COALL-97 patients are first stratified according to HR and LR criteria and then are stratified again according to the PVA score. LR and HR patients with a score of 3 + 4 receive less intensive treatment in reinduction therapy to reduce the cumulative dose of anthracyclines and to mitigate the high rate of infectious complications during this part of the protocol. LR patients with a score of 7-9 are treated according to the HR protocol; HR patients with a score of 8 + 9 receive BMT if an HLA identical family donor is available. In study COALL-97 the results of the minimal residual disease study and day 15 bone marrow will be compared with the PVA score. CONCLUSIONS: In vitro drug sensitivity testing is an independent prognostic factor which allows adjustment of therapy to the individual risk of relapse in addition to the traditional risk factors. It can be assumed that in patients with a favourable resistance profile therapy can be reduced without loss of efficacy and that patients with an unfavourable resistance profile might profit from more intensive therapy. PMID- 10472557 TI - Cellular drug resistance in acute myeloid leukemia: literature review and preliminary analysis of an ongoing collaborative study. AB - Cellular drug resistance is one of the main causes of the frequent ultimate failure of chemotherapy in childhood acute myeloid leukemia (AML). We here summarize the results of a literature review on in vitro drug resistance in childhood AML, focusing on studies using so-called cell culture assays. We also briefly describe some results of an ongoing collaborative study between the Research Laboratory of Pediatric Oncology in Amsterdam (University Hospital Vrije Universiteit) and the German BFM-AML Group. In general, the literature and our preliminary data on in vitro cellular drug resistance in AML are promising in terms of clinical relevance. Cell biological features and clinical response to chemotherapy are related to in vitro drug resistance. However, a large study including multivariate analysis is required to more firmly establish the clinical value of cellular drug resistance testing in childhood AML, and the collaborative study will therefore be continued. Possible applications of cell culture assays include risk-group stratification, rational improvements of current treatment protocols for subgroups of patients based on specific drug resistance profiles, individualised tailored therapy, the study of cross-resistance patterns between drugs, the study of possibilities to modulate or circumvent drug resistance, the study of drug interactions, selection of patients for clinical phase II studies and drug screening. PMID- 10472558 TI - [Primary isolated myeolosarcoma in childhood]. AB - Isolated myelosarcomas are rare first manifestations of acute myeloid leukemia (AML), preceding bone marrow involvement by weeks to months. Seventeen of 654 children observed during the studies AML-BFM 87 and 93 were diagnosed as extramedullar myelosarcomas (2.6%). The predominantly myelomonocytic or monoblastic tumor cells (M4 or M5 according to FAB classification) mainly infiltrated skin (n = 8). Additional tumors were located in mucosa (n = 2), central nervous system (n = 2), orbita (n = 2), bone (n = 1), glandulae parotis (n = 1) and lymph nodes. Due to the initial mild and variable symptoms in some children the diagnostic measurements were delayed and treatment was inadequate. This might be responsible for the high rate of relapse (79%) and the poor outcome. Ten of 17 patients died from disease (estimated survival 0.27 +/- 0.13 compared to AML-BFM 87/93 0.51 +/- 0.03). Suspect skin lesions or tumors should be considered as isolated myelosarcoma of a primary manifestation of AML. An intensive AML-specific chemotherapy is recommended. PMID- 10472559 TI - [Prognosis, treatment completion, and complications in nonresponders in the study AML-BFM87]. AB - BACKGROUND: Nonresponders (NR) are patients (pts.) with no or insufficient response to initial treatment, which may be caused by either initial risk factors or poor therapy realization. In study AML-BFM 87, 49 NR of 307 patients (16%) did not achieve remission until the end of intensive chemotherapy and were analysed to assess the specific contribution of prognostic factors, therapy realization and complications of therapy. THERAPY AND METHODS: Therapy started with an 8-day induction therapy followed by a 6-week consolidation and two 5-day intensification blocks with high-dose cytosine-arabinoside and VP-16. Maintenance therapy was given for a total duration of 1.5 years. To evaluate the impact of treatment intensity in NR, we compared the dose compliance (DC = dose given/intended dose), the dose intensity (DI = dose per time given), the treatment results, and toxicity of the individual therapy phases in responders (CR) and NR. RESULTS: In 19 of 49 NR therapy was stopped before starting intensification blocks. Twenty-six NR received at least one block of intensification, and seven patients between three and six intensification blocks. Six children entered maintenance therapy. Twelve patients received a bone marrow transplant (9 allogeneic, 3 autologous). Six (5 after bone marrow transplantation) of 49 NR are still alive for 64 to 108 months. In nearly all patients induction therapy could be applied according to protocol (mean DC: 98%, range 85%-100%), whereas therapy realization was more difficult in the 2nd phase of therapy (mean DC: 92%, range 12%-113%). Deviations from the protocol in the treatment blocks (changes of dose and/or schedule) were mainly attributable to persistence of blasts (n = 33) and septic complications (n = 24). The mean relative DI of 1.01 was according to protocol. Bleeding and infectious complications in the individual therapy phases varied from 7% to 61%. NR compared to CR patients suffered significantly more often from bleeding during the first and second part of consolidation and from infections during the second part of consolidation. Withdrawal from protocol in NR was mainly due to persistence of blasts (n = 16), followed by bone marrow transplantation or other therapies (n = 13), and sepsis (n = 11). CONCLUSIONS: It is difficult to discriminate between nonresponse associated with blast persistence followed by complications and subsequent discontinuation of therapy and nonresponse due to insufficient therapy in patients with complications. Our analyses revealed that therapy with 2 intensifications according to protocol was feasible in 13 NR. Patients' condition permitting, therapy should not be stopped prematurely, in order to sustain the option of BMT after blast cell reduction. PMID- 10472560 TI - [Neoadjuvant therapy for localized osteosarcoma of extremities. Results from the Cooperative osteosarcoma study group COSS of 925 patients]. AB - BACKGROUND: Owing to twenty years of multicentric interdisciplinary cooperation, the COSS group has been able to collect data on a large group of osteosarcoma patients treated by neoadjuvant therapy. This paper reviews results achieved in patients with localized extremity tumors. PATIENTS AND METHODS: INCLUSION CRITERIA: Registration into a completed neoadjuvant COSS-Study. Histologically confirmed, primary, localized, high-grade, central osteosarcoma of an extremity; age < 40 years; no pretreatment; interval diagnosis to chemotherapy < or = 3 weeks; no severe comorbidity. Chemotherapy: HD-methotrexate +/- doxorubicin +/- cisplatin +/- ifosfamide +/- BCD. Scheduled local therapy: Surgery. RESULTS: 925 evaluable patients from 101 institutions. Median age 15 years, m:f 1.4:1. Primary site: femur 510, tibia 251, humerus 100, fibula 51, other 13. Tumor-size < 1/3 of the involved bone 616, > or = 1/3 304. Definitive surgery in 903/925 cases, 443 limb salvage procedures. Good response (> 90% necrosis) in 469/806 (58.2%) evaluated tumors. Median follow-up for surviving patients: 5.42 years. Actuarial survival after 5 and 10 years: 72.5% (95%-CI 69.3-75.7) and 66.3% (62.5-70.0), relapse-free 62.1% (58.7-65.4) and 59.4% (55.8-63.0). 683/925 alive (601 first remission), 242 deceased (212 tumor progression, 30 other causes). 66.2% (97.3%) of all relapses within 2 (5) years. Prognosis correlates with tumor-size (< vs. > or = 1/3: 69.9% vs. 58.3% at 10 years) and -site (tibia: 74.2%, humerus: 54.5%) and -response (good vs. poor: 78.2% vs. 52.5%) (all p < 0.01). Actuarial 10-year survival by response grading I-VI according to Salzer-Kuntschik 80.9%, 82.8%, 71.1%, 60.7%, 47.7%, 27.3%. COSS-studies with preoperative 4-drug therapy more efficacious than less aggressive protocols. No impact of doxorubicin scheduling (sequential: rapid vs. 48 h-continuous infusion) or cisplatin scheduling (randomized: 5 h vs. 72 h-infusion) on prognosis detected. CONCLUSIONS: Intensive multiagent chemotherapy and delayed surgery for localized extremity osteosarcoma led to excellent oncologic results in the COSS-studies. Tumor-size, -site, and response as well as the intensity of upfront chemotherapy correlated with outcome. Giving doxorubicin and cisplatin by continuous infusions did not result in discernible prognostic disadvantages. PMID- 10472561 TI - [Descriptive epidemiology of Ewing's tumor--analysis of German patients from (EI)CESS 1980-1997]. AB - BACKGROUND: Due to low absolute morbidity rates a reliable descriptive epidemiology of the Ewing family of tumors (EFT) has been mainly provided by large population-based cancer registry reports. To date, only few reports on this topic have been published. PATIENTS: The present report is based on data of 945 German patients who were enrolled into the (EI)CESS therapy studies between 1980 and 1997. It is assumed that under the age of 15 years an almost complete registration of all German patients with Ewing's sarcoma has been achieved. Diagnoses in all patients were ascertained by biopsies. METHODS: In this analysis we looked at the associations between year of first diagnosis, age at first diagnosis, gender, the histological subtypes and the primary skeletal localisation of the tumours. RESULTS: The majority of patients with EFT was male (60%). More than half of the patients developed the disease between age 10 and 19 (57%) while about 20% were younger and almost a quarter was diagnosed after age 20 years. The percentage of patients older than 15 years was higher in men (55%) than in women (45%). The age-standardized annual incidence before age 15 years was estimated as 3 per million, and the cumulative incidence up to that age as about 46 per million. Long bones of the lower extremities (32%, femur alone 16%) and the pelvis were the most common sites of primary localisation. While 70 to 80% of all tumours in the long bones of the extremities were composed of Ewing's sarcomas, about one third in the central skeleton and over 20% in the pelvic bones were Primitive neuroectodermal tumours (PNET). Atypical Ewing's sarcoma accounted for about 10% of tumours in all sites. CONCLUSIONS: The large (EI)CESS database enables the meaningful and reliable description of epidemiological characteristics of the rare occurrence of the Ewing family of tumours. Further analyses of this database seem to hold great promise. PMID- 10472562 TI - [EICESS 92 (European Intergroup Cooperative Ewing's Sarcoma Study)-- preliminary results]. AB - BACKGROUND: Ewing tumor patients' outcome is 50% to 60% with current treatment strategies. Questions concerning toxicity and secondary malignancies are of increasing importance. PATIENTS AND METHODS: 631 patients were registered with the German EICESS study center of the European Intergroup Cooperative Ewing's Sarcoma Study, 369 patients were randomized. Treatment apart from local therapy consisted of 14 courses of Vincristine, Actinomycin D, Cyclophosphamide or Ifosfamide, Adriamycin (Doxorubicin), with or without Etoposide. First results concerning event-free survival (EFS), toxicity, and the rate of secondary malignancies three years after diagnosis are presented. RESULTS: Three year EFS was 0.66 for patients with localized tumors, 0.43 for patients with primary pulmonary/pleural metastases, and 0.29 for patients with other metastases, respectively. Large tumor volume or pelvic site, especially if inoperable, were adverse prognostic factors. Both histological and MRT-defined response were positively correlated to outcome. Up to 67% of patients experienced WHO grade IV toxicity, mostly related to bone marrow depression. The treatment related mortality was 1% (6/631). Until 15.02.1999, six of 687 patients have suffered secondary malignancies, two of six after (additional) myeloablative therapy. CONCLUSIONS: EICESS 92 treatment is toxic, but manageable and compares favorably to international results. New strategies must be sought for certain risk groups of patients. PMID- 10472563 TI - [High-dosage chemotherapy in primary metastasized and relapsed Ewing's sarcoma. (EI)CESS]. AB - BACKGROUND: Patients (pts) with primary metastatic Ewing tumours (ET) have a poor prognosis for event free survival (EFS) compared to pts with localised disease. Following relapse the prognosis is extremely poor. Therefore these primary metastatic and relapsed pts were piloted for high dose therapy (HDT) for the last years. PATIENTS AND METHODS: Between April 1984 and May 1997, 131 ET pts who underwent HDT were registered in the German CESS/EICESS office: 79 pts with primary metastases and 52 pts with relapsed tumours. After induction therapy, consisting of chemotherapy and local therapy, pts received high dose regimens, mainly based on melphalan and/or etoposide (92%). Stem cell rescue was applied from allogeneic bone marrow (n = 13), autologous bone marrow (n = 17), or peripheral blood stem cells (n = 95). The date of analysis was September 1st, 1998. Outcome was calculated by Kaplan-Meier-analyses. RESULTS: The median time under study since high dose therapy was 3.7 years. 35/131 pts (26.7%) were in continuous complete remission, 80/131 pts (61.1%) had relapsed or progressed, 11/131 pts (8.4%) died of complications and 5/131 pts (3.8%) presented with secondary malignancies. For the total group of primary metastatic pts, EFS five years after diagnosis was 19% for pts with HDT and 27% for those without (p = 0.9209). The subgroup of pts with primary lung and bone metastases seemed to benefit from HDT (EFS five years after diagnosis: 34% versus 5%, p = 0.0001). Outcome of pts with an early ET relapse (< 2 years) was also improved by HDT (EFS four years after relapse: 17% versus 2%, p = 0.0001). CONCLUSIONS: The total group of primary metastatic ET pts showed no obvious benefit from HDT, based on melphalan and/or etoposide. Pts with metastases to multiple organ systems, and early relapse seemed to benefit from HDT. The value of HDT should be assessed in prospective clinical trials. PMID- 10472564 TI - [Chemotherapy in fibromatoses of childhood and adolescence: results from the Cooperative soft tissue sarcoma study (CWS) and the Italian Cooperative study group (ICG-AIEOP)]. AB - BACKGROUND: Fibromatoses are a group of semimalignant tumors which grow infiltratively without metastases. If radical surgery is not at all possible or only by mutilation the question of neoadjuvant concepts arises. PATIENTS: We report on therapy and outcome of 36 patients registered to the German CWS study group and the Italian ICG study group for soft tissue sarcoma who were between 0 and 23 years of age. 15 had a histology of infantile (myo-) fibromatosis (9 unifocal, 6 multifocal). 15 patients had desmoid-like, aggressive fibromatosis. METHODS: We performed a retrospective analysis of the patients registered to the CWS and ICG study group. For literature review a medline search was performed. RESULTS: 23 patients received chemotherapy with anthracyclines (VAIA) or without (VAC) or monotherapy with alkylating agents. In 19 cases response to chemotherapy was measurable. In 9 patients tumors responded to chemotherapy. In some cases response was evident only after 20 weeks of treatment. In the group of non responders treatment was stopped early (after 4 weeks) in some cases. Relapses occurred mainly in the group of desmoid-like, aggressive fibromatosis (7 out of 15). In the group of infantile (myo-) fibromatosis only one patient suffered a relapse (1 out of 9). The literature review showed that mainly ADR/DTIC, VAC and MTX/VBL was used for inoperable fibromatoses. Concerning the response rates there where no major differences. Concerning toxicity the regimen with methotrexate and vinblastine seems superior to the others. CONCLUSIONS: As a chemotherapy regimen of first choice vinblastine with low dose methotrexate can be recommended. The therapy should not be stopped before the 20th week of treatment. The histological subgroup can provide good information on the relapse rate and aggressivity to be expected. PMID- 10472565 TI - [Radiation therapy in juvenile aggressive fibromatosis]. AB - BACKGROUND: In about one third of patients suffering from a desmoid tumor primary complete resection is not feasible. Furthermore in locally relapsing tumors reoperation alone does not result in cure in many cases. Radiotherapy can be applied in both groups of patients with curative intention. But the indication of radiotherapy is challenging particularly in children and adolescents due to the impending late radiation sequelae such as growth delay, fibrosis and radiation induced secondary malignancy. PATIENTS AND METHOD: The follow up and outcome of five irradiated children/adolescents with desmoid tumors, registered in the German-Cooperative-Soft-Tissue-Sarcoma Study (CWS) was looked at, and the corresponding literature was reviewed. RESULTS: Radiotherapy of gross residual or relapsing tumors resulted in long lasting event free survival in two cases (3/8 years), but in one patient local progression occurred despite irradiation. Postoperative radiotherapy in patients with microscopic residual disease resulted in both, long lasting event free survival (14 years, 1 patient) and in early local relapse (1.5 years, 1 patient). The role of radiotherapy could not be evaluated clearly by the CWS-experience due to the fact that the irradiated patients were treated individually also by chemotherapy and/or tamoxifen. But despite sparse and retrospective data there is evidence in the literature, that radiotherapy is able to control 65-90% of the unresectable desmoid tumors and that the local relapse rate can be reduced by radiotherapy by 10-20% in patients with microscopic residual disease following resection. CONCLUSIONS: Radiotherapy as primary treatment should be given if complete tumor resection is not feasible without mutilation. Radiotherapy can be applied postoperatively if the risk of local relapse seems to be highly life- or function threatening. PMID- 10472566 TI - Testicular germ cell tumors, an update. Results of the German cooperative studies 1982-1997. AB - BACKGROUND: Oncologic treatment of childhood testicular germ cell tumors can be regarded as a model of curable neoplasm. Over 50% of the tumors are stage I A, produce alpha-fetoprotein and thus provide after semicastration a "wait and see" policy. PATIENTS: The MAHO 82, 88, 94 cooperative studies registered between 1982 and 1997 197 patients, 110 patients had yolk sac tumors (YST), 47 differentiated teratomas (TD), 38 malignant teratomas of either intermediate (MTI), undifferentiated (MTU), or trophoblastic type (MTT) and two seminomas. After semicastration only 65 patients received standard chemotherapy according to stage and histology consisting of four courses of vinblastine, bleomycin and cisplatin. If after two courses viable tumor was indicated, delayed laparotomy was performed (seven patients). Patients with incomplete tumor response after two courses received three courses of etoposide, ifosfamide and cisplatin (nine patients). RESULTS: 105 patients had YST stage I, five higher stages of disease. One of these died by tumor progression. Of 91 patients followed according to "wait and see" only 14 needed standard chemotherapy. The NED of 105 patients is 99%. 47 patients had TD stage I; the NED is 100%. 13 patients had malignant teratomas stage I. 13 patients had stage II and received chemotherapy; the NED for these 26 patients is 100%. 12 patients had stages III or IV, four died. CONCLUSION: In testicular germ cell tumors of childhood in alpha-fetoprotein producing tumors of stage I A a "wait and see" program is safe. X-irradiation or primary lymphadenectomy can be omitted since chemotherapy alone reveals excellent results. PMID- 10472567 TI - Analysis of treatment efficiency of carboplatin and etoposide in combination with radical surgery in advanced and recurrent childhood hepatoblastoma: a report of the German Cooperative Pediatric Liver Tumor Study HB 89 and HB 94. AB - BACKGROUND: Hepatoblastoma (HB) is the most common liver tumor of childhood, and comprises approximately 1% of all pediatric malignancies. Although recent data from multicenter trials of GPOH, SIOP, CCG and POG indicate a remarkable improvement of therapy results, the prognosis of advanced or recurrent HB is still not satisfying. PATIENTS AND METHODS: During 1989 and 1997, the German Cooperative Pediatric Liver Tumor Studies HB 89 and HB 94 registered 141 patients with HB, who were treated according to the study protocols. These patients received standard chemotherapy with ifosfamide, cisplatin and doxorubicin (IPA) pre-operatively and/or post-operatively. Fourteen children with recurrent or advanced HB were additionally treated with carboplatin and etoposide (CARBO/VP 16), the reason being observations of drug resistance in children with HB after four or more courses of IPA-therapy in the HB 89 study. The clinical data and course of these patients were evaluated to investigate the efficiency of CARBO/VP 16 chemotherapy and for analyzing the role of surgery. RESULTS: Mean follow-up for survivors was 4.3 years (range 13 months-8 years). Tumor resection was attempted in 13 children but, in only 3 cases, was a complete tumor resection achieved in one operation. There was no perioperative death, and 7 of the patients (50%) are in remission. Two patients underwent adjuvant chemotherapy with CARBO/VP 16 for advanced HB at first operation: all are alive and well. Five patients with local relapse and/or distant metastases responded partially to CARBO/VP 16 therapy, and a complete remission was achieved in one patient. In five patients, progressive disease was observed during therapy with CARBO/VP 16. One patient, stable while on chemotherapy, had a successful resection. Acute toxicity of chemotherapy was observed in 7 patients (50%). CONCLUSION: An aggressive approach using IPA and CARBO/VP 16 chemotherapy and highly developed surgical techniques may improve the prognosis of advanced or recurrent HBs. PMID- 10472568 TI - [Cytomegalovirus infection as a possible factor in the progression of neuroblastoma]. AB - BACKGROUND: There is evidence that the infection with human cytomegalovirus is clinically associated with enhanced metastasis and progression of neuroblastoma disease. An in vitro model with HCMV-infected neuroblastoma cells (NB) was used to investigate whether HCMV modulates the metastatic potential of NB. METHODS: The neuroblastoma cell line UKF-NB-4 and its productively and persistently HCMV infected variant UKF-NB-4AD169 were cocultured with human endothelial cells (EC). The rate of NB adherent to the endothelial monolayer and the rate of transmigrating NB was determined by means of combined reflexion interference contrast/phase contrast microscopy. RESULTS: UKF-NB-4AD169 adhered to and transmigrated through cocultured EC monolayer to a significantly higher extent compared with the non-infected cell line UKF-NB-4. At the cell-to-cell contact sites between UKF-NB-4AD169 and EC the intercellular endothelial contacts loosened resulting in the formation of reversible focal openings in the monolayer. This phenomenon was not observed with UKF-NB-4. The transendothelial migration rate of UKF-NB-4AD169 was therefore significantly higher than that of UKF-NB-4. The formation of focal openings in the endothelial monolayer and the enhanced transmigration rate of UKF-NB-4AD169 was suppressed in the presence of phenantroline, suggesting that HCMV-induced proteinases might be responsible for this phenomenon. CONCLUSION: The results confirm our assumption that HCMV has the ability to modulate functional properties of NB which are essential for the interactions with endothelial cells and thus for metastasation. The clinical relevance of these findings has to be further defined yet by means of prospective studies with HCMV-infected neuroblastoma patients. Proteinase inhibitors could be valuable in the therapeutic treatment of these patients. PMID- 10472569 TI - [Natural human IgM-antibodies in neuroblastoma therapy: preliminary findings of a phase I/II clinical trial]. AB - BACKGROUND: Sera from healthy individuals contain natural IgM antibodies (anti-NB Ab) cytotoxic to neuroblastoma (NB) cells. In contrast to healthy children the prevalence of anti-NB-Ab in sera of NB patients is low. Binding of anti-NB-Ab to the NB cell surface leads to activation of complement in vitro. In vivo the injection of the purified IgM fraction from a cytotoxic blood donor results in complete growth arrest of NB xenografts in nude rats. Preliminary results from a phase I/II study to evaluate the pharmacokinetics and side effects of a therapy with anti-NB-Ab are presented here. PATIENTS: Included in this study are patients with NB stage 4 according to INSS with relapse or non-responders to therapy according to the GPOH-NB-therapy protocol. The patients are negative for anti-NB Ab and are older than one year. METHODS: The therapy is based on a complete exchange of the anti-NB-Ab negative patient serum against serum from an anti-NB Ab positive ABO-compatible donor by means of plasmapheresis. RESULTS: Up to now, 14 cycles of plasmapheresis have been carried out in 6 NB patients. In 13 of 14 therapy cycles a significant increase in serum toxicity could be observed. Severe side effects have not been seen except a catheter associated thrombosis which was reversible under heparin treatment. After plasmapheresis, pain in the tumor site or regions of metastasis did occur regularly. In some cases temporary elevation of body temperature occurred. One patient had a reduced MIBG uptake after therapy. Tumor necrosis was observed in 2 patients. Three patients showed tumor progress. CONCLUSION: Immunotherapy of NB in children by serum exchange using anti-NB-Ab positive ABO-compatible donor serum is feasible without major side effects and leads to a significant increase of serum toxicity against NB cells. PMID- 10472570 TI - Betulinic acid: a new chemotherapeutic agent in the treatment of neuroectodermal tumors. AB - We identified betulinic acid (BetA) as a new cytotoxic agent active against neuroectodermal tumor cells including neuroblastoma, medulloblastoma, glioblastoma and Ewing's sarcoma cells representing the most common solid tumors of childhood. BetA induced apoptosis independent of wild-type p53 protein and accumulation of death-inducing ligand/receptor systems such as CD95. BetA had a direct effect on mitochondria resulting in the release of soluble apoptogenic factors such as cytochrome c or AIF from mitochondria into the cytosol where they induced activation of caspases. Overexpression of the anti-apoptotic proteins Bcl 2 or Bcl-XL that blocked loss of the mitochondrial membrane potential and cytochrome c release from mitochondria conferred resistance to BetA at the level of mitochondrial dysfunction, protease activation and nuclear fragmentation. Neuroblastoma cells resistant to CD95- or doxorubicin-triggered apoptosis remained sensitive to treatment with BetA suggesting that BetA may bypass some forms of resistance. Moreover, BetA exhibited potent antitumor activity on primary tumor cell cultures from all neuroblastoma (4/4), all medulloblastoma (4/4) and most glioblastoma patients (20/24) ex vivo. These findings suggest that BetA may be a promising new agent in the treatment of neuroectodermal tumors in vivo. PMID- 10472571 TI - Valproic acid for the treatment of pediatric malignant glioma. AB - Despite surgery and adjuvant cytotoxic therapy anaplastic astrocytoma, glioblastoma and diffuse intrinsic brain stem glioma continue to have dismal prognosis. Differentiation induction is a new approach taking into account that malignant glioma cells share many features with immature glial progenitor cells that are capable of terminal differentiation. The concept of differentiation therapy is currently evaluated for several pediatric malignancies with or without multimodal standard therapy. Valproic acid (VPA) is a branched chain fatty acid that is able to inhibit proliferation of neuroectodermal cells and to induce these cells along neuronal or glial lineage. Preclinical studies have shown that VPA inhibits growth of human and rodent glial tumor cells in vitro and induces a distinct mature glial phenotype. In addition, growth of human neuroblastoma cells is inhibited in vitro and in vivo and exhibits marked evidence of differentiation. Treatment of neuroblastoma and glioma cells with VPA was accompanied by changes of surface molecule expression that enhance immunogenicity and reduce their capability to metastasize. The antitumoral effects observed in preclinical studies were reached at concentrations that are readily achieved in patients treated with VPA for epilepsy. Epilepsy patients receiving VPA have significantly enhanced hemoglobin F levels, supporting the hypothesis that nontoxic levels of VPA can induce cellular differentiation. Broad clinical experience with VPA and its low toxicity encourage the evaluation of VPA in patients that have been submitted to postoperative combined chemo- and radiotherapy for pediatric malignant glioma. PMID- 10472572 TI - Fanconi anemia and beta c deficiency-associated pulmonary alveolar proteinosis as two hereditary diseases of childhood which are potentially curable by stem cell gene therapy but require different therapeutic approaches. AB - Fanconi anemia (FANC) and pulmonary alveolar proteinosis associated with deficiency of the beta-chain common to the GM-CSF/IL3/IL5 receptors (beta c-PAP) are rare inherited disorders of childhood or adolescence. Hematopoietic stem cell gene therapy aiming at reintroducing the wildtype cDNA as a new concept for the treatment of hereditary diseases may be applicable to FANC and PAP, as both disorders can be successfully treated by allogeneic stem cell transplantation. However, there are important distinctions to be made between the two diseases: FANC seems to be a disorder with functional stem cell deficiency. Thus, introduction of the wildtype cDNA should provide an in vivo growth advantage to genetically corrected stem cells so that corrected cells and their progeny may expand in vivo and slowly repopulate the entire hematopoietic system. In beta c PAP, the defect has no major impact on proliferation or differentiation of stem cells. Therefore, introduction of the wildtype gene will probably not provide any selective growth advantage and the percentage of corrected cells in the hematopoietic compartment depend on the percentage of stem cells initially transduced as the current technology only allows for transduction of stem cells with low efficiency. The introduction of a second selectable cDNA into the vector might be used to provide selective growth for modified cells and thus overcome a low gene transfer efficiency of stem cells. The correction of rare monogenetic diseases may serve as a model for gene therapy prior to attempts to treat more common and complex polygenetic diseases. The studies outlined here will be helpful envisioning new treatment strategies for other inherited monogenetic diseases such as mucopolysaccharidosis, Gauchers disease or adrenoleukodystrophy. PMID- 10472573 TI - Tumor vaccines--application to childhood cancer. AB - Currently, several clinical studies explore the therapeutic potential of tumor vaccines which are genetically modified to produce immunostimulatory molecules as a complementary approach for conventional cancer therapy. In this review the immunological basis and the preclinical design of such vaccine strategies are described with particular emphasis to acute leukemia and neuroblastoma. The role of cytokines, chemokines and costimulatory surface molecules for generation of tumor vaccines is summarized, and the advantages and disadvantages of autologous, allogenic and dendritic cell vaccines are discussed. Finally, combination immunogens are introduced as a potent means of enhancing the anti-tumor response. PMID- 10472574 TI - [Antiinfectious prophylaxis in pediatric oncology. Work group "Quality Assurance" of Society for Pediatric Oncology and Hematology (GPOH)]. AB - Infections in disease- and/or chemotherapy-related neutropenia are major, often emergency-type problems in the treatment of pediatric oncology patients and explain the ongoing discussion about antiinfectious prophylaxis. The different aspects of prophylaxis and an overview on the literature are presented. Antiinfectious prophylaxis in pediatric oncology includes the following issues: 1. General aspects such as information for patients and parents on neutropenia and risk of infectious diseases and indication and management of reverse isolation and barrier isolation; 2. antibacterial prophylaxis with oral non absorbable and oral absorbable antibiotics; 3. Pneumocystis carinii (Pc) prophylaxis; 4. antifungal prophylaxis to prevent disseminated candidiasis and aspergillosis; 5. antiviral prophylaxis, especially varicella-zoster-virus (VZV) post-exposure prophylaxis and cytomegalovirus (CMV) prophylaxis; 6. immunoglobulins and hematopoietic growth-factors (HGF); 7. active immunization. An evaluation of those measures leads to the following conclusions: A major controversy exists regarding antibacterial and antifungal prophylaxis. Probably not effective are the use of reverse isolation and of oral, non-absorbable antibiotics. Oral absorbable antibiotics, antifungal prophylaxis using fluconazole and amphotericin B and the use of hematopoietic growth factors are likely to be effective. Clearly effective are strict hand-washing procedures, Pc and CMV prophylaxis and passive vaccination against VZV in case of VZV exposure of a seronegative patient. PMID- 10472575 TI - Recancostat compositum therapy does not prevent tumor progression in young cancer patients. AB - Stimulated by the public discussion about an antimalignant effect of the supplement drug Recancostat compositum (200 mg glutathione, 40 mg cysteine, 50 mg anthocyanin) the drug was given in high doses to 16 young patients with intensively pretreated and measurable solid tumors for 1-30 weeks. In all patients the disease progressed, and they died 1-48 weeks following the start of the therapy. Side effects were not seen. This observation does not support the claim of an antitumor effect of Recancostat compositum. PMID- 10472576 TI - Late cardiotoxicity of anthracyclines in children with acute leukemia. AB - Congestive heart failure is one the most severe late complications of cancer therapy with anthracyclines. The function of the heart muscle was evaluated in 50 children (30 boys and 20 girls), aged from 5 years 6 months to 20 years 7 months, treated in the past for lymphoblastic or nonlymphoblastic acute leukemia. The total dose of the administrated anthracyclines was 120-550 mg/m2. The circulatory system was evaluated on the basis of history, physical examination, ECG, exercise test and echocardiography. Impaired contractility of the heart muscle was found in 32% of cases. The degree of impairment was related to the total dose of anthracyclines and to the period from discontinuation of therapy. Heart muscle function disorders were present also in children, in whom the cumulative dose of anthracycline antibiotics did not exceed 400 mg/m2. In the majority of patients the evidence of heart damage was subclinical. PMID- 10472577 TI - [AIDS: the myth of Sisyphus]. PMID- 10472578 TI - [Difficulties in the management of diabetes in medically underserved countries: the example of Madagascar]. PMID- 10472579 TI - [Discovery of a focus of intestinal bilharziasis in te Republic of Djibouti]. AB - An unprecedented pocket of intestinal schistosomiasis was discovered in the Republic of Djibouti in 1997. The first cases were diagnosed in French and Djiboutian tourists who presented initial symptoms of bilharzian infection after bathing in the fresh-water basin under Hassan Gari Bira Falls, near Randa. Seventeen cases were subsequently confirmed by detection of anti-schistosome antibodies using indirect hemagglutination (IH) and indirect immunofluorescence (IIF) and/or detection of Schistosoma mansoni eggs in the stool. Further testing was performed in 35 village inhabitants, mostly children, who had been exposed by bathing in the basin. The IH reaction was positive in 28 patients (80 p. 100) including 17 (49 p. 100) with levels greater than 1/64. In 92 p. 100 of cases, IH findings were confirmed by IIF which indicated that association with hypereosinophilia was common. Schistosoma mansoni eggs were found in stools from 7 patients (19 p. 100) who generally displayed mild hypereosinophilia. Information concerning the zone of risk was distributed and control measures were undertaken as widely as possible in Djibouti and abroad. PMID- 10472580 TI - [Malacologic-schistosomal study of the refugee camps on the Plain of Ruzizi in the Democratic Republic of the Congo]. AB - Bilharziasis due to Schistosoma mansoni is widespread in the eastern part of the People's Republic of the Congo, particularly in the Ruzizi plain area. The intermediate host is the fresh-water snail Biomphalaria pfeifferi whose ecology is now well documented. Bilharziasis due to Schistosoma haematobium and Schistosoma intercalatum have not yet been observed but the potential for this development is high given the presence of the intermediate hosts, i.e. Bulinus truncatus, Bulinus globosus, and Bulinus forskalii, and the migration of populations in the area. Malacological and cercariometric studies were undertaken in 3 refugee camps in 1995. Findings showed that Biomphalaria pfeifferi was present in all water systems tested but that levels varied from 21 p. 100 to 70 p. 100 from one camp to another. The rate of infestation by Biomphalaria pfeifferi varied from 37 to 20 p. 100 depending on the sampling site. Given the prevalence of Schistosoma mansoni in the population, extension of bilharziasis is likely in the Ruzizi plain area and its surroundings. Potential intermediary hosts for Schistosoma haematobium and Schistosoma intercalatum were of lower relative prevalence and were found in specific biotopes. The authors propose methods of controlling water snails and preventing infection of people and water systems. PMID- 10472581 TI - [Rapid resolution of Plasmodium ovale malarial attacks using artesunate (Arsumax)]. AB - This randomized, non-comparative clinical trial without placebo was carried out to assess the efficacy and tolerance of artesunate for treatment of acute Plasmodium ovale malarial attacks. Thirty Cameroonese patients were included. All presented acute Plasmodium ovale malarial attacks with parasitemia in excess of 500 asexual forms per mm3. Four days after treatment with artesunate, all 30 patients were asymptomatic with no parasitemia. Reduction rates were 93.9 p. 100 for asexual forms and 75.4 p. 100 for gametocytes. Parasite clearance was achieved within 38.8 hours and fever disappeared within 36.6 hours. Tolerance was excellent in 29 patients. The remaining patient briefly complained of mild vertigo. A transient decrease in reticulocyte levels was observed in one patient initially presenting anemia. Artesunate appears to achieve rapid and complete resolution of acute Plasmodium ovale malarial attacks. Since artesunate eliminates both asexual forms and gametocytes, it also acts on transmission by limiting the duration of survival of asexual forms. PMID- 10472582 TI - [Salmonellosis in HIV infection in a hospital setting in Gabon]. AB - A case-control study was carried out in Libreville, Gabon, to determine the incidence of salmonella infection in HIV patients and identify any special clinical, therapeutic, or prognostic features. The records of 3000 patients hospitalized in the Infectious Disease Department of the Jeanne Ebori Foundation between January 1990 and December 1994 were studied. The incidence of salmonella infection, serotype, clinical presentation, prognostic factors and therapeutic modalities were compared in 2759 HIV-positive patients and 441 HIV-negative patients. Salmonella infection was noted in a total of 208 patients (58 HIV positive and 150 HIV-negative). The incidence of salmonella infection was 13 p. 100 (non-typhoid in 76 p. 100 of cases) in HIV-positive patients versus 5.4 p. 100 in HIV-negative patients. The predominant serotypes in HIV-positive patients were Salmonella typhimurium and Salmonella enteritidis which accounted 41 p. 100 and 26 p. 100 of the isolated strains. The only significantly difference in clinical presentation was a higher incidence of bacteremia (84 p. 100) and focal lesions (11.4 p. 100) in HIV-positive patients with low-grade salmonella infection. The duration of treatment was three weeks with cotrimoxazole and 10 days with fluoroquinolones and cephalosporines. The outcome in HIV-positive patients was recurrence-free cure in 40 p. 100, multiple relapses in 12 p. 100, and death in 24 p. 100. The remaining patients were lost from follow-up. This study demonstrates the gravity of low-grade salmonella infection in African HIV positive patients. Early diagnosis is necessary to allow chemotherapy which can be effective despite immunodeficiency. PMID- 10472583 TI - [The prevalence of schistosomiasis in Togo. A cross-sectional study conducted in a school setting]. AB - As a prelude to a national campaign to control schistosomiasis in Togo, mass screening of school children in 22 prefectures was undertaken to determine the extent of endemic schistosomiasis. Children were randomly selected for testing. In each case, stool examinations using by the Kato-Katz method and urine tests (centrifugation of 10 ml) were performed to detect Schistosome eggs. A total of 2511 children were tested. The sex ratio was 1.7 and mean age 10.4 years (range: 5 to 20 years). The incidence of schistosomiasis was 26.7 p. 100. Schistosoma haematobium was the most widespread and active Schistosome species, being endemic in all locations studied. The incidence of Schistosoma haematobium ranged from 0.6 to 72 p. 100 (national mean: 25.5 p. 100). Schistosoma mansoni was prevalent in only 12 prefectures (range 0.6 to 10 p. 100; national average 2.2 p. 100). The rate of infection was highest in the age group between 10 and 20 years and the risk appeared to be significantly higher in males than in females (p < 0.05). This study confirmed the high incidence of schistosomiasis infection in Togo and identified hyperendemic areas in which control measures should be programmed first. PMID- 10472584 TI - [A malarial attack on return from a voyage to the French Antilles. Discussion of the mode of transmission]. AB - Malaria has been considered to be eradicated from the French West Indies for over 25 years. In this report we describe a patient who was hospitalized and successfully treated in Paris for severe Plasmodium falciparum malaria after returning form a brief trip to Guadeloupe. Several modes of transmission are possible. Given the presence of a reservoir of Plasmodium falciparum gametocytes in the Haitian immigrant community and persistent breeding of Anopheles albimanus in the French West Indies, the most likely explanation is local transmission. This is the second case of malaria involving travelers to Guadeloupe to be reported within the last ten years and the first time that autochtonous transmission has been considered. Falciparum malaria should be included in differential diagnosis for patients presenting fever after returning from travel in the French West Indies, a highly popular tourist destination. PMID- 10472585 TI - [Fatal fulminating hepatitis due to Herpes simplex virus type 2 in a young immunocompetent female]. AB - Fulminant herpes simplex viral hepatitis is uncommon in immunocompetent subjects. A 24-year-old woman presenting hepatomegaly with fever was hospitalized after returning from a trip to southern Africa. The patient was neither pregnant nor immunocompromised. Because of recent tropical travel, differential diagnosis included alphabetic hepatotropic virus infection, yellow fever, African hemorrhagic fever, and arbovirus infection. After ruling out other common viral etiologies, a definitive diagnosis of herpes simplex viral infection was made on the basis of clinical and laboratory findings showing high fever, leukopenia, and thrombopenia; of histological examination of the native liver after transplantation showing non-inflammatory confluent focal hemorrhagic necrosis; and on serologic tests demonstrating seroconversion for herpes simplex virus type 2. Outcome after transplantation was rapidly fatal but the death was not directly related to infection. The most likely etiology of fulminant hepatitis in a young woman returning from travel in a tropical area is hepatitis virus B or hepatitis virus E in cases involving pregnancy. However herpes simplex virus should be included in differential diagnosis even in immunocompetent subjects. PMID- 10472586 TI - [Rigid endoscopy and laryngo-tracheo-bronchial foreign bodies in children: observations apropos of 200 endoscopies conducted in a tropical setting]. AB - Between 1986 and 1998, 200 rigid bronchoscopic procedures under general anesthesia were carried out at the Principal Hospital in Dakar, Senegal for foreign body extraction from the distal airways of 194 children. For the study period, the incidence of this accident was 3.7 p. 1000. Sixty-three percent of patients were male and 77 p. 100 were under 4 years of age. Most patients (69 p. 100) were examined within 48 hours after the accident. Examination of clinical records showed that aspiration was mentioned during anamnesis in only 56 p. 100 of cases. Persistent coughing (80 p. 100) and mild dyspnea (70 p. 100) were the most common symptoms. Auscultation of the lungs was negative in 25 p. 100 of cases and anterior x-ray of the neck and chest were normal or poorly informative in 59 p. 100. In 154 of the 200 procedures, extraction of the foreign body was successful from the trachea in 35 p. 100 of cases, the larynx in 13 p. 100, the right main stem bronchus in 31 p. 100 and the left main stem bronchus in 21 p. 100. In the remaining 46 cases, extraction was unsuccessful. The most frequent foreign body was a peanut. No deaths occurred in this series but cardiac arrest was observed in 6 patients during or immediately after endoscopy. This experience confirms the indication for immediate rigid bronchoscopy in cases involving aspiration or persistent respiratory symptoms. The high incidence of this accident suggests that information campaigns should be undertaken in health care facilities, households, and schools. PMID- 10472587 TI - [Neisseria meningitidis and meningitis]. AB - Meningococcal meningitis epidemics can occur anywhere in the world. However this risk is particularly high during the dry season in the sub-Saharan zone of Africa known as the Lapeyssonnie meningitis belt. This area characterized by hyperendemicity that regularly gives rise to epidemics. Multilocus enzyme electrophoresis has made possible identification and monitoring of the progression of virulent clones of Neisseria meningitidis strains in the world. Monitoring is now possible by multilocus sequence typing and data bank on the Internet. Vaccination is a major prophylactic modality. The usefulness of plain group A plus C polysaccharide vaccines is limited because of poor effectiveness in young children who constitute the highest risk group. During epidemics, mass vaccination should be carried out as early as possible according to the state of alert defined for the area. More recent conjugate vaccines against group A and C, which are effective in young children and provide long-term protection by induction of immunologic memory, may allow routine vaccination in the future. Although clinical signs are often apparent, not all cases are diagnosed by clinical examination unless gravity is taken into account. Untreated the disease is always fatal. The only hope of survival is early institution of appropriate antimicrobial therapy (even prior to hospitalization). Several strains resistant to chloramphenicol have been reported and the number of strains with reduced sensitivity to penicillin is rising constantly. Although treatment remains feasible, the existence of resistant forms raises the need to monitor the sensitivity of meningococci using standardized of antibiograms. PMID- 10472588 TI - [Human American trypanosomiasis 90 years after its discovery by Carlos Chagas. II -Clinical aspects, physiopathology, diagnosis and treatment]. AB - Cardiac, neurologic, and gastrointestinal manifestations of Chagas Disease have been well documented, but underlying pathophysiologic mechanisms, especially of chronic myocarditis, remain unclear. In the last decades, vectorial and transfusional transmission has been diminished. However congenital forms, which were long unknown and are still poorly understood, and reactive forms, which occur in patients with acquired or induced immunodeficiency, have confounded conventional wisdom concerning the evolution of this disease. Several new diagnostic tools have been developed without replacing the traditional methods, e.g. the xenodiagnostic technique proposed by Brumpt. With regard to indirect diagnosis, further progress is needed to improve specificity and sensitivity as well as the discriminating ability of the numerous techniques now available. Recently several double-blind randomized trials showed that benznidazole may be useful for early stage disease in children under 12 years of age. Further study with long-term follow-up will be necessary to determine the value of generalizing treatment to all patients with Chagas disease regardless of age and disease stage. However the ideal trypanocidal agent has yet to be found. Although attempts to immunize animals have not been complete failures, current results are not adequate to hold forth hope of a vaccine for use in man within the foreseeable future. PMID- 10472590 TI - [The resurgence of Chagas disease in French Guiana: a challenge to be answered]. PMID- 10472589 TI - [Health assessments of a military company stationed on the Maroni River in French Guiana]. AB - Over a 5 month period (October 1996 to February 1997), a rotating company of 146 servicemen belonging to the Navy Airborne 6th Regiment were assigned along the Maroni River in French Guyana. During this mission, the medical personnel treated 387 local residents. Etiologies comprised 51 malaria attacks including 46 involving Plasmodium falciparum and 4 rattlesnake envenomations. The most common cause of consultation by military personnel was mycotic and staphylococcal skin infections, but 5 cases involving poor acclimatization were treated during the hot and dry season. Seven malaria attacks involving Plasmodium falciparum including 2 that were severe occurred despite prophylaxis using chloroquine proguanil. Treatment with halofantrine was successful in all but one case which required combined chemotherapy using quinine and doxycycline. Five cases of cutaneous leishmaniasis were observed in subjects involved in jungle training. No case of HIV infection was detected upon returning home since most personnel either followed the recommendation to abstain from sex (51 p. 100) or used a condom (90 p. 100 of personnel who had sexual relations). These data illustrate the health risks for mainland French nationals in the region of the Maroni River and underline the need for preventive measures and education. PMID- 10472591 TI - [Indigenous strongyloidiasis: apropos of a case]. PMID- 10472592 TI - [Gas gangrene after injection of quinine salts: apropos of 2 severe pediatric cases]. PMID- 10472593 TI - [Impact on health of early supplementation with rice in infants in Laos]. PMID- 10472594 TI - [Cardiac sarcoidosis in the Principal Hospital in Dakar: apropos of a case]. PMID- 10472595 TI - [Survival and the quality of life in extrahospital cardiorespiratory arrest]. AB - BACKGROUND: There are few data in Spain on out-of-hospital cardiac arrest and the efficacy of emergency systems. The objectives of the present study were to evaluate an emergency system, comparing survival at hospital discharge according to the origin, cardiac or non-cardiac, of cardiac arrest in out-of-hospital critically ill patients, and to describe the quality of life of the survivors. PATIENTS AND METHODS: Prospective study on 282 patients treated during 1995 and 1996 by ICU ambulances units of the Andalusian Public Health Emergency Company (061) in Granada, Almeria and El Ejido-Poniente (Spain). The Utstein style was followed, gathering the mortality at different times up to 6 months after hospital discharge and the origin (cardiac/non-cardiac) of the arrest. Quality of life 6 months after discharge was collected among survivors. RESULTS: Advanced cardiopulmonary resuscitation (CPR) was applied to 176 (62.4%) patients. The survival rate to discharge was 4.9% for patients with cardiac etiology (7/142 x 100) and 5.9% for those with non-cardiac etiology (2/34 x 100), with non significant differences between the two groups. An optimal quality of life in all domains, except for pharmacological dependence in seven, was found in the eight survivors 6 months after hospital discharge. CONCLUSIONS: The origin (cardiac/non cardiac) of out-of-hospital cardiac arrest is not associated with survival at hospital discharge. The survivors exhibit an optimal quality of life 6 months after discharge. PMID- 10472596 TI - [A randomized comparative study of 3 days of azithromycin treatment and 10 days of cefuroxime treatment in exacerbations in patients with chronic obstructive pulmonary disease]. AB - BACKGROUND: The aim of this study was to prospectively evaluate the clinical and gasometric evolution and the side effects of two treatment schedules in the exacerbations of patients with chronic obstructive pulmonary disease (COPD): 500 mg/24 h of azithromycin (AZM) for three days versus 500 mg/12 h of acetyl cefuroxime (ACF) for 10 days. PATIENTS AND METHODS: Patients were randomized included into each therapeutic schedule. The patients were seen three times (days 1 and 4, and at 15-21 days) to evaluate clinical symptoms scores. Forced spirometry and arterial gasometry were performed the first and the last time the patients were seen. The number of patients requiring admission during follow up and the secondary effects of each antibiotic were quantified. RESULTS: A total de 50 patients were treated with AZM and 51 with ACF. The evolution of the symptoms was similar although with a trend to greater improvement in those treated with AZM. This improvement was significant for the characteristics of expectoration (p < 0.05). Functional and gasometric evolution was similar in the two schedules. Three patients treated with AZM required hospital admission, as did 5 treated with ACF. A greater number of secondary effects were observed in patients treated with ACF (18%) than in those receiving AZM (10%), with gastrointestinal side effects being the most commonly observed. CONCLUSIONS: Treatment with short schedule of AZM may have the same activity as longer schedule of ACF, with fewer secondary effects thereby suggesting that AZM may be an effective alternative in the treatment of exacerbations in patients with COPD. PMID- 10472597 TI - [The presence of minimal residual disease during induction therapy is a reliable factor for the prognosis of recurrence in children with standard-risk B-cell precursor acute lymphoblastic leukemia]. AB - BACKGROUND: To evaluate the prognostic implications of the presence of minimal residual disease (MRD) during the induction period in children diagnosed with low risk B cell precursor acute lymphoblastic leukaemia (LR-ALL). PATIENTS AND METHODS: In 36 children with LR-ALL treated with BFM-86 protocol, MRD was studied at the end of induction by IgH and TCR delta gene analysis. RESULTS: The probabilities of disease free survival were: 52% (patients with MRD at the end of Induction), 64% (patients with MRD at day 15) and 100% (patients without MRD). CONCLUSIONS: The presence of MRD is a new and useful prognostic factor in LR childhood ALL. PMID- 10472598 TI - [Cardiorespiratory arrest and afterwards what?]. PMID- 10472599 TI - [Should H. pylori infection in duodenal ulcer be confirmed before administering eradicating treatment?]. PMID- 10472600 TI - [How to improve the statistical quality of articles presented to biomedical journals: a checklist for authors]. PMID- 10472601 TI - [The mechanisms of Mycobacterium tuberculosis disease and the implications for its prevention in patients with HIV infection]. PMID- 10472602 TI - [Mydriasis due to accidental contact with stramonium (Datura stramonium)]. PMID- 10472603 TI - [The number of patients that it will be necessary to treat: a tool of interest for the clinician]. PMID- 10472604 TI - [The information to be given by a resident physician for correct consent]. PMID- 10472605 TI - [The information to be given by a resident physician for a correct consent]. PMID- 10472606 TI - [An intrathoracic hypertension syndrome or spontaneous pneumomediastinum?]. PMID- 10472607 TI - [The autologous transplantation of hematopoietic precursors on an outpatient basis: an analysis of its feasibility at the Hospital Clinic de Barcelona]. AB - BACKGROUND: Autologous stem cell transplantation (ASCT) is increasingly performed on an outpatient basis. Preliminary results show a significant financial saving without modifying the therapeutic results. The aim of this study has been to evaluate the feasibility of this procedure in our the Hospital Clinic of Barcelona, Spain. PATIENTS AND METHODS: All patients in whom an ASCT due to a hematological malignancy, having a good performance status, a caregiver 24 h per day, a lodging near to the hospital and an appropriate cultural and social environment, were offered an outpatient management starting the day after stem cell administration (day +1). RESULTS: During the study period (April, 1997-June, 1998) 64 ASCT were performed. Fifteen patients (23%) were managed on an outpatient basis. The remaining patients were not included in the study due to patient preference (26%), medical reasons (22%), distant home (18%) and social problems (9%). Seven patients (47%) had to be readmitted in the hospital due to fever (n = 4), nausea (n = 2) or respiratory distress (n = 1). Patients receiving total body radiation (TBI) in the conditioning regimen did not have a higher readmission rate than those conditioned without TBI. The median hospital stay (since day +1) was 6 (range: 0-22) and 17 (13-117) days for outpatient and inpatients respectively (p = 0.0016). This reduction would imply a saving of 65% in hospitalization charges and of 12% in total charges. All patients having elected the outpatient management were very satisfied by having made that choice. CONCLUSION: With an appropriate patient selection, outpatient ASCT is a feasible method that may improve the quality of life of the patients and can reduce the costs of the procedure. PMID- 10472608 TI - [The prognostic value of cytomegalovirus antigenemia and viremia for the development of cytomegalovirus disease and the survival of AIDS patients]. AB - OBJECTIVES: To analyse risk factors for morbidity and survival associated with blood cytomegalovirus (CMV) detection with the antigenemia method among AIDS patients. PATIENTS AND METHODS: CMV antigenemia and CMV blood cultures in 277 AIDS patients IgG-CMV sero-positive with a CD4 level lower than 200 x 10(6)/l under antiretroviral monotherapy were analysed. We consider cases the 116 patients with one or more positive blood samples tested for pp65 antigenemia or CMV culture. They were matched with 161 control patients with negative antigenemia or viremia. RESULTS: Multivariate analysis pointed out a significant positive association for blood CMV reactivation with the following variables: CMV disease development and CMV urine detection, sex-acquired HIV infection, CD4+ < 50 x 10(6)/l and matched time from AIDS diagnosis to CMV blood culture correlated with positive antigenemias. Quantitative antigenemia title showed predictive value for risk of CMV disease although 23% of retinitis patients had persistent undetectable antigenemia. CMV invasive disease developed in 48% of cases and 11% of controls (relative risk [RR]: 7.9; 95% confidence interval [CI]: 4.2-14.7). Mortality after 12 months of follow-up was 73% vs 52% respectively (p < 0.001). Time survival curves after CD4+ count adjusting remained significantly lower for case patients (median, 127 days vs 355 days; p < 0.01 by log-rank test). Increased death rate was found in patients with CMV disease (74%), followed by patients with CMV antigenemia but no disease (70%) and patients without antigenemia or CMV disease (mortality 49%). CONCLUSIONS: CMV blood detection in AIDS patients may be considered as a bad prognosis marker for CMV morbidity and survival. This risk increases with higher CMV antigenemias. Therefore, pre emptive anti-CMV therapy should be considered in this restricted population. PMID- 10472609 TI - [Progressive multifocal leukoencephalopathy associated with human immunodeficiency virus infection: the clinical, neuroimaging, virological and evolutive characteristics in 35 patients]. AB - BACKGROUND: The clinical, neuroimaging, virologic and evolutive characteristics of progressive multifocal leukoencephalopathy (PML) in 35 AIDS patients are studied. PATIENTS AND METHODS: PML was diagnosed by clinical and neuroimaging criteria in 32 patients and by autopsy in other three. The detection of JC virus (JCV) was done by PCR and further hybridization of the amplified DNA in peripheral blood lymphocytes, urine and CSF. RESULTS: 127 of 930 HIV positive patients were admitted by neuropsychiatric symptoms and of them 35 (SD 27.6%) by PML. The PML patients had a mean CD4 lymphocytes count of 75.3 (82.0)/x 10(6)/l and a HIV viral load of 330,698 (538,971) copies of RNA/ml. Thirty patients did not receive any anti-retroviral therapy or only transcriptase inhibitors monotherapy and five triple anti-retroviral therapy, including a proteases inhibitor. Multiple hypodense lesions on CT (53.1%) and T2 hyperintense lesions on MRI (58.3%) were the most frequent neuroimaging findings. JCV was detected in 20/21 (95.2%) LMP patients: 18/19 detections in lymphocytes, 6/8 in CSF and 4/6 in urine. The mean survival without and with antiretroviral therapy were 3.0 (0.47) and 21.4 (4.4) months (p < 0.001) in 34 patients followed. PML progressed to death in 31/34 patients (91.2%), and remained stable in 3/34 (8.8%). A patient was lost for follow-up. CONCLUSIONS: The application of clinical and neuroimaging criteria and the detection of JCV in CSF are useful for high presumption diagnosis of PML without brain biopsies. JCV detection in lymphocytes and in urine have a much lower predictive value. The evolution and survival of this disease can improve with triple anti-retroviral therapy including a protease inhibitor. PMID- 10472610 TI - [Liver transplantation in patients infected with the human immunodeficiency virus: the difficult challenge of a new stage]. PMID- 10472611 TI - [Is the medical prescription of heroin a treatment option for heroin addiction? The PEPSA Team (The Experimental Program of Narcotic Prescription in Andalucia]. PMID- 10472612 TI - [The role of the epidermal growth factor in cell and tissue protection]. PMID- 10472613 TI - [Fever, abdominal pain and coma with a rapidly unfavorable evolution in a 48-year old patient with acute promyelocytic leukemia during the period of post chemotherapy pancytopenia]. PMID- 10472614 TI - [Acute hepatitis due to ritodrine]. PMID- 10472615 TI - [Neuroleptic malignant syndrome associated with olanzapine]. PMID- 10472616 TI - [Health-related absenteeism among workers employed in various work environments]. AB - The state of health in working people is determined by a number of factors among which working conditions and the kind of job performed play undoubtedly a crucial role. The observation and analysis of trends in sick absenteeism may provide indirect information on health problems of the occupationally active population. The aim of the analysis presented was to define the magnitude and the causes of temporary work disability among workers employed in individual branches of the national economy, and finally to identify high risk groups of workers because of their employment in specific working conditions. The statistical material used as the basis for the calculation of absenteeism indicators embraced the data on the number of work disability days, derived from a 15 representative sample of punched cards, concerning certificates of temporary work disability. In order to accomplish the objectives of the analysis, the trends in sick absenteeism observed before 1989, the period preceding the structural changes and the transformation of Polish economy were taken into consideration. The analysis indicates that the level of sick absenteeism in the branches of the national economy (forestry, building construction, transport, industry) and the proportion of chronic diseases in its structure prove the effect of working conditions on the workers' health. Considering individual branches of industry, the highest levels of sick absenteeism are observed in the following branches: mining for men (5.61), non-ferrous metals for women (10.04) and men (4.95), ferrous metallurgy for women (9.99) and men (4.93), and petroleum products for women (11.76). The analysis of sick absenteeism in the branches selected in view of hazardous working conditions shows that: (1) in many branches chemical hazards in the work environment are responsible for an increased sick absenteeism due to diseases of the circulatory system, particularly among men, mental disorders and neoplasms, both among men and women, and complications of pregnancy among women; (2) physical work overload increases sick absenteeism mainly due to diseases of the musculoskeletal and peripheral nervous systems, as well as due to cardiac disease and arterial hypertension; and (3) hot microclimate contributes to sick absenteeism because of diseases of the circulatory system, including cardiac disease and arterial hypertension, particularly among men, as well as acute and chronic respiratory diseases. PMID- 10472617 TI - [The levels of health education among diabetic patients of the Lublin railroad health service on the basis of adequate self-awareness]. AB - The studies were carried out in the years 1995-97. From the group of 1000 diabetic patients (railway workers) of the Lublin Railway Health Service, 160 persons were chosen at random. They were interviewed in order to recognise their knowledge of the disease they suffered from and which could contribute to the prevention of diabetes in this occupational group. The studies revealed that the desired knowledge was lacking among the diabetic railway workers who did not show any interest in increasing their awareness in this regard. PMID- 10472618 TI - [Sources of finance for provincial occupational health services. Theory and practice]. AB - The financing of occupational health services (OHS) at the provincial level is an important issue in view of the transformation process going on not only in OHS but also in the overall health care system in Poland. New principles of financing must be now based on the cost and effects analyses. Thus, the question arises on how to provide financial means adequate to needs of health care institutions resulting from their tasks and responsibilities. The gaps existing in the information system have encouraged us to examine the situation in regard to the structure of financing and internal allocation of financial means. The objectives were formulated as follows: to characterise the sources of financial means received by provincial OHS centres; to analyse the structure of financial means derived from various sources, taking into account forms of financial administration, using the data provided by selected centres; to define the relation between the financial means being at the disposal of OHS centres and the scope of their activities; The information on the financing system was collected using a questionnaire mailed to directors of selected OHS centres. The information collected proved to be a valuable source of knowledge on the above mentioned issues as well as on how far the new system of financing associated with a new form of financial administration--an independent public health institution--has already been implemented. The studies indicated that at the present stage of the OHS system transformation it is very difficult to formulate conclusions on the financing administration in provincial OHS centres. PMID- 10472619 TI - [The labor potential in occupational health services at the provincial level before the occupational health services act came into effect]. AB - The author evaluates how far the personnel of occupational health services (OHS) at the provincial level is ready to undertake and implement the tasks provided by the Occupational Health Services Act at the eve of its coming into force. Personnel resources of OHS differ depending on the region. This applies to all groups of workers and is generally related to the provincial regulations and employment policy. Specialists-consultants constituted 12% of provincial OHS workers and the majority of them (60%) were employed on a full-time basis. Specialists in laryngology, ophthalmology and neurology were employed most frequently. Physicians with additional qualifications necessary for performing preventive examinations accounted for 16% of the total number of OHS workers; and those with acknowledged competence due to long experience--40%. Thus, 60% (400) of physicians should be trained in research institutions. As to qualifications of nurses, the situation is even worse. Only 6% of nurses were specialised in occupational health, and 35% of nurses employed at the provincial level completed relevant qualification courses. The qualifications of over 60% of nurses are insufficient for implementing tasks provided by the new OHS Act. PMID- 10472620 TI - [Problems in the evaluation of miners' exposure to radon in Poland in view of international organizations recommendations and European Union directives]. AB - Exposure of miners to natural radiation in which radon-222 plays the major role has been studied in Poland since the end of the nineteen sixties. The work environment measurements and personal monitoring methods have been developed for monitoring the exposure. A quite wide range of doses resulting from big differences in radon concentrations in mines is characteristic of miners' exposure. It was estimated that about 16% of miners received doses above 5 mSv per year. This group of miners should be provided with individual dosymentry. In this paper methodological and legal aspects of the situation in mines are analysed. A large number of Polish recommendations and legal regulations must be changed in order to harmonise them with the directives of the European Union. PMID- 10472621 TI - [Chromium carcinogenicity]. AB - Chromium belongs to the group of trace elements hich are essential in numerous functions of the human body. Chromium deficiency may be responsible for various dysfunctions, whereas exposure to chromium at higher concentrations is toxic and may lead to the occurrence of neoplastic diseases. Epidemiological studies of chromium exposure proved its carcinogenity, and thus the IARC recognised Cr(VI) and its compounds as one of ascertained carcinogens. Some findings of these studies were reviewed in the first part of this work. The second part presents some molecular aspects of chromium carcinogenity which are still the subject of medical research. The direct and indirect effects of chromium and its compounds on DNA are analysed as are the relationships between the level of chromium oxidation and carcinogenity, and between the presence of reductants and the kind of DNA damage. Methods for the assessment of chromium mutagenity and genotoxicity are also discussed, and special attention is paid to tests of mutation in bacteria and yeast as well as to sister chromatid exchange (SCE) test. PMID- 10472622 TI - [Industrial wastes and health hazards]. AB - Uncontrolled industrial waste sites are potential sources of unplanned release of hazardous substances into the environment. According to the reports of the State Inspectorate for Environment Protection (PIOS), 60% of hazardous waste sites registered in Poland create potential or real threat to public health and the environment. hazardous substances migrating off the site can spread and affect the environmental media: ground water, surface water, air, soil, biota. They can also enter the human body. As a result of research studies aimed at identifying the hazard and assessing health effects posed by hazardous waste sites in the United States, the Agency for Toxic Substances and Disease Registry (ATSDR) has identified seven groups of health effects most common among populations living in the vicinity of waste sites. These are: birth defects, cancer, immune function disorders, kidney and liver dysfunction, lung and respiratory diseases and neurotoxic disorders. However, the evidence based on exposure-effect relationship is not generally very strong. Most of the studies lack sufficiently accurate exposure assessment. PMID- 10472623 TI - [The department of occupational medicine of the State School of Occupational Medicine at the National Institute of Occupational Medicine in Warsaw in 1926 1939]. AB - The Department of Occupational Hygiene, State School of Hygiene at the National Institute of Hygiene was established in 1926/27. Prof. Brunon Nowakowski was its founder and the first Director. Teaching and providing of highly qualified personnel for occupational medicine institutions was the main area of the Department's activity. The Department also carried out scientific research focusing on organisation of labour hygiene in Poland, and occupational diseases, lead poisoning in particular. The results of studies carried out by the Department provided the basis for the elaboration of numerous legal regulations in the area of occupational medicine. PMID- 10472624 TI - Formation of glycolaldehyde phosphate from glycolaldehyde in aqueous solution. AB - Amidotriphosphate (0.1 M) in aqueous solution at near neutral pH in the presence of magnesium ions (0.25 M) converts glycolaldehyde (0.025 M) within days at room temperature into glycolaldehyde phosphate in (analytically) nearly quantitative yields (76% in isolated product). This robust phosphorylation process was observed to proceed at concentrations as low as 30 microM glycolaldehyde and 60 microM phosphorylation reagent under otherwise identical conditions. In sharp contrast, attempts to achieve a phosphorylation of glycolaldehyde with cyclotriphosphate ('trimetaphosphate') as phosphorylating reagent were unsuccessful. Mechanistically, the phosphorylation of glycolaldehyde with amidotriphosphate is an example of intramolecular delivery of the phosphate group. PMID- 10472625 TI - Does formate reduce alpha-ketoglutarate and ammonia to glutamate? AB - The reported reduction of alpha-ketoglutarate and ammonia by formate is much slower than described (Morowitz et al., 1995). The formate reduction if any is small under these conditions. Glutamate is produced from a reduction by a second molecule of alpha-ketoglutarate involving an oxidative decarboxylation. PMID- 10472626 TI - Pyrite suspended in artificial sea water catalyzes hydrolysis of adsorbed ATP: enhancing effect of acetate. AB - Minerals have been implicated in different catalytic processes during chemical evolution. It has been proposed that exergonic synthesis of pyrite (FeS2) could have served to promote the endergonic synthesis of biomonomers in early stages of life formation on Earth. The present study was aimed to investigate whether pyrite can adsorb nucleotides and oxo acids in the potentially mild prebiotic conditions found away from the hot hydrothermal vents. It is shown that pyrite strongly adsorbs adenosine 5'-triphosphate in an artificial medium that simulates primordial aqueous environments, and that adsorption is enhanced in the presence of acetate and in an oxygen-free atmosphere. Moreover, the mineral catalyzes the sequential hydrolysis of the gamma and beta phosphoanhydride bonds of the nucleotide. PMID- 10472627 TI - The fidelity of template-directed oligonucleotide ligation and the inevitability of polymerase function. AB - The first living systems may have employed template-directed oligonucleotide ligation for replication. The utility of oligonucleotide ligation as a mechanism for the origin and evolution of life is in part dependent on its fidelity. We have devised a method for evaluating ligation fidelity in which ligation substrates are selected from random sequence libraries. The fidelities of chemical and enzymatic ligation are compared under a variety of conditions. While reaction conditions can be found that promote high fidelity copying, departure from these conditions leads to error-prone copying. In particular, ligation reactions with shorter oligonucleotide substrates are less efficient but more faithful. These results support a model for origins in which there was selective pressure for template-directed oligonucleotide ligation to be gradually supplanted by mononucleotide polymerization. PMID- 10472629 TI - Habitability of planets around red dwarf stars. AB - Recent models indicate that relatively moderate climates could exist on Earth sized planets in synchronous rotation around red dwarf stars. Investigation of the global water cycle, availability of photosynthetically active radiation in red dwarf sunlight, and the biological implications of stellar flares, which can be frequent for red dwarfs, suggests that higher plant habitability of red dwarf planets may be possible. PMID- 10472628 TI - The antibiotic viomycin as a model peptide for the origin of the co-evolution of RNA and proteins. AB - Viomycin is an RNA-binding peptide antibiotic which inhibits prokaryotic protein synthesis and group I intron self-splicing. This antibiotic enhances the activity of the ribozyme derived from the Neurospora crassa VS RNA, and at sub-inhibitory concentrations it induces the formation of group I intron oligomers. Here, we address the question whether viomycin exerts specificity in the promotion of RNA RNA interactions. In an in vitro selection experiment we tested the ability of viomycin to specifically select molecules out of an RNA pool. Group I intron RNA was incubated with a pool of random sequence RNA, or with a pool of RNA molecules which had previously been enriched for viomycin-binding RNAs. Viomycin was added in order to select viomycin-binding RNAs and to guide their interaction with the intron RNA resulting in recombinant molecules. Viomycin was indeed capable of specifically selecting RNA molecules which contain viomycin-binding sites promoting recombination. These results suggest that small peptides are able to play the role of selector molecules in a putative 'RNA World' launching the co evolution of RNA and proteins into an 'RNA-protein World'. PMID- 10472630 TI - Directed molecular evolution. AB - We propose the existence of a relationship of stereochemical complementarity between gene sequences that code for interacting components: nucleic acid-nucleic acid, protein-protein and protein-nucleic acid. Such a relationship would impose evolutionary constraints on the DNA sequences themselves, thus retaining these sequences and governing the direction of the evolutionary process. Therefore, we propose that prebiotic, template-directed autocatalytic synthesis of mutally cognate peptides and polynucleotides resulted in their amplification and evolutionary conservation in contemporary prokaryotic and eukaryotic organisms as a genetic regulatory apparatus. If this proposal is correct, then the relationships between the sequences in DNA coding for these interactions constitute a life code of which the genetic code is only one aspect of the many related interactions encoded in DNA. PMID- 10472631 TI - [The Review of Respiratory Diseases on the verge of the XXIst century]. PMID- 10472632 TI - [Dyspnea and panic attacks]. AB - Patients with panic attacks (or acute anxiety episodes) often present respiratory symptoms, especially dyspnea. Biological, behavioral and cognitive models of panic attacks are presented. The "false suffocation alarm model" is outlined. Biological and cognitive-behavioral treatments which use, among other techniques, respiratory regulation and hyperventilation, are described. Controlled studies and several meta-analyses show that cognitive-behavioral therapies represent an alternative to anxiolytics and antidepressants in panic attacks with or without agoraphobia, and have long lasting effects, while most of the patients under medication alone relapse when pharmacological treatments are stopped. PMID- 10472633 TI - [Anti-infective aerosols]. AB - Anti-infectious agents such as pentamidine, antibiotics (mainly colistine and aminoglycosides) and amphotericin B can be administered by aerosol. This route of administration is not officially approved and it constitutes an empirical approach which has benefited from recent research which is summarized hereafter. The most fundamental question is related to the potentially deleterious effects of nebulization processes, especially ultrasound, on the anti infectious properties of the drugs. Colimycin, which was chosen as a reference because its polypeptide structure makes it unstable a priori, proved to be resistant to high frequency ultrasound, which is encouraging for other molecules such as aminoglycosides or betalactamins. The nebulizer characteristics have also to be taken into account. An aerosol can be produced from an amphotericin B suspension and from colistine using both an ultrasonic nebulizer and a jet nebulizer. Distinction between good and bad nebulizers does not depend upon the physical process involved to nebulize the drug, but on the intrinsic characteristics of the device and its performance with a known drug. The inhaled mass of an aerosol in the respirable range must be high and dosimetric nebulizers represent a significant progress. Finally, adminnistration of anti infectious aerosols requires a new pharmacological approach to monitor treatment and urinary assays are promising. PMID- 10472634 TI - [Autonomic nervous system and obstructive sleep apneas]. AB - Understanding of the pathophysiology of obstructive sleep apnoea, a common yet relatively newly recognised condition, has advanced rapidly in recent years. This condition produces major acute haemodynamic changes and causal relationships with arterial hypertension and cardiovascular morbidity have been proposed. The role that the autonomic nervous system plays in mediating these cardiovascular changes has been the focus of intensive research activity and the development of new techniques in physiological monitoring, such as spectral analysis of heart rate variability, Finapres blood pressure monitoring, measurement of muscle sympathetic nerve activity, radionuclide tests and animal models of obstructive sleep apnoea have substantially increased the knowledge base. The acute haemodynamic changes are associated with high levels of sympathetic discharge and with fluctuating parasympathetic activity. There are also chronic changes in baroreceptor and chemoreceptor reflexes associated with an increase in baseline daytime sympathetic activity and abnormal vagal reflex responses to voluntary respiratory manoeuvres. These acute autonomic changes appear to be provoked by a combination of stimuli triggered by hypoxaemia, upper airway responses, ventilatory changes and arousal. The mechanisms of the chronic autonomic changes are less clear; it is likely that recurrent hypoxaemia is important, but the roles of recurrent ventilatory stress and arousal are not clear. Normalising respiration with CPAP therapy prevents the acute cardiovascular changes and reduces the acute sympathetic over-activity, and in compliant patients, restores abnormal vagal responses to normal and reduces excess chronic sympathetic activity. Whether or not this produces a reduction in long term cardiovascular morbidity is not established. PMID- 10472635 TI - [Series on "respiratory resuscitation" published under the aegis of the Resuscitation Group of the French-Language Society of Pneumology]. PMID- 10472636 TI - [History of respiratory resuscitation]. PMID- 10472637 TI - [Scope of and perspectives on resuscitation and pneumology]. PMID- 10472638 TI - [The relationship between intensive care medicine and pneumology in Switzerland. The situation in 1998]. AB - We describe the relationships in Switzerland between two medical specialties, critical, or intensive, care and the respiratory medicine. To date, there are only few direct relationships between these medical fields, and very rarely a respirologist becomes the head of a critical, or an intensive care unit. There are two main reasons explaining this situation. First, internal medicine, or general internal medicine, is still a strong specialty in Switzerland, and the patients are frequently under the direct responsibility of an internist, whereas the pneumologists remain consultants. Second, intensive care medicine is a full specialty, distinct from anaesthesiology, with a strong structure. However, things are moving, and the general tendency now is to reduce the total number of beds in the Swiss hospitals, and to open some specialised wards under the direct supervision of specialists, including pneumologists. Nevetherless, due to the small size of the country and the important number of decentralised hospital structures, specialised intensive care units, as respiratory intensive care units, are unlikely to be open in the future. PMID- 10472639 TI - [Respiratory resuscitation in Belgium]. PMID- 10472640 TI - [Respiratory resuscitation in Quebec. General aspects and implications of pneumologists in intensive care units in "New France"]. AB - ICU set up is a complex framework in Quebec. In this respect, quebecer and french systems are very different. Pulmonologists are one of the most committed sub specialists in ICUs, either as consultant or as MD on duty. Amongst paramedics, respiratory therapists are essential members of the team. Invasive and noninvasive ventilations are commonly performed. Critical care teaching program and structures for developing and supporting clinical research activities are in place. PMID- 10472641 TI - [Pneumology and resuscitation. A typical American marriage?]. PMID- 10472642 TI - [Physiopathology of cystic fibrosis lung disease]. AB - Cystic fibrosis is a common genetic disorder caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Although the link between the mutations of the CFTR gene and the progressive lung disease is not completely understood, noteworthy advances have been made in the understanding of the mechanisms underlying lung injury in CF. The pathophysiologic cascade starts with CFTR dysfunction resulting from mutations of the CFTR gene, and leading to abnormal ion and water transport across airway epithelia. Abnormal airway surface liquid may in turn lead to impairment of mucociliary clearance, airway mucosal anti-microbial defenses, chronic bacterial infection and inflammation. Each of these steps has been elucidated in more detail in the recent years. This is of relevance from a therapeutic viewpoint. New therapeutic interventions include gene therapy, agents that may circumvent the defect in intracellular processing of mutated CFTR, agents that may modulate defects in epithelial ion transport, and anti-inflammatory agents. PMID- 10472643 TI - [Fertility and cystic fibrosis. Management at pregnancy]. AB - Life expectancy of patients with cystic fibrosis has regularly increased in recent years, currently exceeding the age of 30 years. The disease now concerns adolescents and young adults more than children. Thus problems linked to fertility have to be considered by physicians caring for adult patients. In males, sterility is quite constant. Changes are related to bilateral agenesis of the efferent ducts leading to excretory azospermia which is one of the major diagnostic criteria leading to the diagnosis of cystic fibrosis in adults. This form of sterility may be relieved by removal of sperm from the epididymis to achieve in vitro fertilization. In the female, fertility is sometimes diminished either by disease-specific abnormalities (tubal epithelium and cervical mucus) or impaired by respiratory failure or nutritional factors. Pregnancy is however possible and is observed in an increasing number of cases. Pregnancy does not appear to alter the general prognosis of the disease as long as cardiorespiratory function is preserved. In all cases, a prognostic evaluation and genetic advice are indispensable. PMID- 10472644 TI - [Coping strategies utilized by asthma patients]. AB - In the Lazarus' transactional model of stress, each individual when facing a stressful situation, set up specific adjustement strategies called coping, including a meaningful pattern of cognitive, behavioral, emotional and somatic responses. Coping strategies used by asthmatic patient are relatively unknown. The objective of this study was to assess the usual coping strategies. We administered the WCC (a shortened version of Lazarus and Folkman's questionnaire) to 116 asthmatic patients and 880 healthy adults French subjects (males and females). A principal component analysis, followed by varimax rotations yielded three factors accounting for about 68.2% of the total variance. They were interpreted as Problem-focused, Emotion-focused and Social-support seeking types of coping. These two first dimensions of coping were closed to those generally described in the literature. Asthmatic patients used more emotion-focused strategy than the control group. These results and their theoretical and practical implications are discussed. PMID- 10472645 TI - [Neurological examination and brain computed tomography in the initial staging of non-small-cell lung cancer: a prospective study]. AB - Brain metastases occur in 17 to 40% of lung carcinoma and 30 to 60% of brain metastases originate from a lung carcinoma. Brain metastasis directly influences prognosis and treatment of lung cancer. The aim of this study was to prospectively compare the findings of the neurological examination performed by a neurologist and results of double dose delayed computed tomography (CT DDD). The neurologist and radiologist were blinded to each other's results. Patients included had non-small-cell lung cancer (NSCLC) and were neurologically asymptomatic with no other cancer. From November 1993 to May 1996, 135 patients were included (126 men and 9 women). Ninety neurological examinations were normal, 34 suggested brain metastasis and 11 were abnormal but did not suggest brain metastasis. One hundred thirteen CTs were normal, 1 showed a brain metastasis and 11 were abnormal but did not evidence brain metastasis. The sensitivity, specificity, positive predictive value and negative predictive value of the neurological examination were 73, 79, 23 and 97% respectively. The presence of brain metastasis was directly related to tumor stage but not to age or histology. We suggest that brain CT DDD should be performed in stage IIIA, IIIB, IV whereas in stage I or II, the neurological examination is sufficient. However, a larger number of patients would be required to confirm these findings. PMID- 10472646 TI - [Chest wall tumors. Report of 17 cases]. AB - The aim of this work was to evaluate the different approaches to surgical repair of the thoracic wall and to discuss technical indications. From June 1987 to June 1997, we cared for 17 patients, 14 males (82.3%) and 3 females (17.7%) with parietal neoplasia. All patients underwent a preoperative respiratory work-up to identify tumoral extension. In 6 patients, the morphology and location of the tumor led to CT-guided transthoracic needle aspiration. Tumoral excision in 14 patients (82.3%) included wide resection of osteomuscular structures. Reconstruction of the thoracic wall associated myoplasty in all cases. A prosthesis was installed in 5 cases and a rib transposition in 2. Pathology examination of the surgical specimen revealed 13 primary tumors (76.5%) and 4 secondary tumors (23.5%) CT-guided transthoracic needle aspiration confirmed the diagnosis in 82.2% of the cases. Twelve patients (70.5%) were alive and recurrence free at 85.6 +/- 40 months after surgery. Five patients died (29.5%) 12.2 +/- 10.1 months after surgery. There was one case of prosthesis infection (5.8%). The appropriate choice of the surgical technique and repair materials gave satisfactory oncological, esthetic and functional results independently of the extent of the parietal defect. PMID- 10472647 TI - [Mucoepidermoid bronchial tumor with an unusual radiologic presentation]. AB - We report a case of low-grade malignant mucoepidermoid bronchial tumor in a 21 year-old woman who presented with hemoptysis. There was a striking radiological presentation with a unilateral clear radiograph. Lung scintigraphy demonstrated perturbed ventilation and perfusion of the left lung. Pulmonary angiography only showed diminished peripheral vascularization. The main vessels were clear. These anomalies fit the radiological category of air trapping accompanied by probably hypoxic pulmonary vasoconstriction. Mucoepidermoid tumors account for only 0.5-1% of all primary lung tumors. They are found in adults as well as in children. There are two clear-cut histological forms with very different prognosis. The tumors of high-grade malignancy are considered as undifferentiated carcinomas and should be treated as such. The prognosis is much better for tumors of low-grade malignancy and current treatment is surgical in the absence of recurrence after isolated endoscopic treatment. PMID- 10472648 TI - [Occupational asthma caused by a coloring reagent]. AB - A case of occupational asthma associated with rhinitis and conjunctivitis occurred in a patient working in a textile factory and exposed to different coloring reagents is reported. Symptoms appeared in phases correlated with changes in the use of the coloring reagents. Prick tests were carried out with the different coloring reagents and were positive for the yellow color. The test was followed by a clear increase in bronchial hyperreactivity to methacholine. Some individual preventive measures, such as wearing a face mask, led to reduction then disappearence of the symptoms. PMID- 10472649 TI - [Endodermal pulmonary tumor resembling a fetal lung]. AB - We report a case of a 56-year-old man who developed a flu-like syndrome associated with weight loss. Chest X-ray revealed a homogeneous round lesion of the lung with sharply defined margins in the central lobe. At pathology, the tumor was found to be an endodermal pulmonary tumor resembling a fetal lung. This uncommon tumor is generally a fortuitous chest X-ray finding seen as a rounded peripheral opacity. Diagnosis is based on pathology examination of the surgical specimen. Complementary investigations are not contributory. Unlike blastoma, a differential diagnosis, prognosis is generally good. PMID- 10472650 TI - [Pleuro-pulmonary blastoma. Report of 4 cases]. AB - Pleuropulmonary blastoma is an uncommon malignant lung tumor observed in children. Outcome is often unfavorable. Two boys and two girls, mean age 4.5 years, were admitted for nonspecific respiratory signs. Oriented by radiology findings, the diagnosis of pleuropulmonary blastoma was confirmed at pathology examination of a pneumonectomy specimen. Three of the children were given postoperative adjuvant chemotherapy. There were three deaths and one child was lost to follow-up. We discuss the clinical features of pleuropulmonary blastoma. No optimal treatment has been defined for this often fatal tumor. PMID- 10472651 TI - [Paraneoplastic hypercorticism associated with a bronchial adenocarcinoma]. AB - Cushing's syndrome is infrequently associated with adenocarcinomas of the lung. We present the clinical features of this syndrome in one case report. The pathogenesis of the syndrome explains the clinical signs, rather different from classical Cushing's disease and also the highly suggestive biological features. Ketoconazole improves clinical signs and biological abnormalities when etiological treatment is not effective. PMID- 10472652 TI - [A case of febrile hemoptysis]. PMID- 10472653 TI - [Suspicious pulmonary opacity and shoulder pain]. PMID- 10472654 TI - [Portable electronic spirometry with peak flow and FEV1 memory]. PMID- 10472655 TI - [Silicosis and scouring powder]. PMID- 10472656 TI - [Autosomal dominant nocturnal frontal lobe epilepsy: the syndrome]. AB - The identification of the autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) in 1994 was rapidly followed by that of other familial forms of non lesional partial epilepsies (familial temporal lobe epilepsy, autosomal dominant partial epilepsy with variable foci). Since then around forty families with ADNFLE have been described, most of them having only 3 or 4 affected individuals. The epilepsy usually begins during childhood (mean age at onset: 11 years). The seizures mainly consist of motor elements which can be dystonic, tonic or hyperkinetic (bipedal automatisms, pelvic thrashing movements...), often preceded by a non specific aura. They are brief and frequent, taking place at night, in clusters. Some patients also present some diurnal seizures. One third of the patients report the occurrence of rare secondarily generalized tonic-clonic seizures. There is a large intra-familial variability as to age of onset, intensity of the manifestations and the course of the epilepsy. During the period of highest frequency of seizures, some patients may present moderate neuropsychological disturbances concordant with frontal dysfunctioning, or transitory behavioral disorders. The seizures often subside with age and may even disappear at adulthood. The most effective antiepileptic drug is carbamazepine, however pharmacoresistance is seen in 20 to 30 p. 100 of the cases. Interictal EEG shows non specific epileptiform anomalies with a frontal predominance, often seen solely on sleep recording, in more than half of all patients. Ictal EEG does not always give evidence of definite ictal discharges. The clinical heterogeneity of ADNFLE as it is especially observed in very variable types of auras which are non localizing, aside form the EEG's own limits, makes it difficult to localize the primary epileptic focus with certainty in the frontal lobe in all cases. In all, the clinical and electrical spectrum of ADNFLE is large, and the topographical identification of these familial frontal lobe epilepsies sets the same problems as for sporadic, classical cryptogenic frontal lobe epilepsies. PMID- 10472657 TI - Gene defects in idiopathic epilepsy. AB - Idiopathic epilepsies are mainly due to genetic factors. In most cases the mode of inheritance is either oligogenic or multifactorial. Only a few rare idiopathic epilepsies are single gene disorders. Monogenic epilepsies offer the chance to identify genes/gene families which might also be involved in the aetiology of common forms of the disease. The genetic basis of two monogenic epilepsies have recently been identified: autosomal dominant nocturnal frontal lobe epilepsy and benign familial neonatal convulsions. PMID- 10472658 TI - Is it possible to anticipate seizure onset by non-linear analysis of intracerebral EEG in human partial epilepsies? AB - Detection of electrophysiological features preceding and indicative of imminent seizures in patients with epilepsy would be a major breakthrough with immense scientific and clinical implications. The definition of a "pre-ictal state" several minutes prior to seizure onset would open a new time window for studying mechanisms of seizure generation as well as for possible therapeutic interventions. In this review we present recent findings from nonlinear time series analysis of intracranially recorded EEG that may allow to forecast seizure onset in patients with partial epilepsy. PMID- 10472659 TI - Neuronal nicotinic acetylcholine receptors: their properties and alterations in autosomal dominant nocturnal frontal lobe epilepsy. AB - Identification of genes coding for the neuronal nicotinic acetylcholine receptors (nAChRs) has allowed rapid progress in the field of neuroscience. Determination of a high-affinity binding site for nicotine that correlates with the expression of mRNAs coding for nAChRs as well as protein expression is the best demonstration for localization of these receptors. Reconstitution of functional nAChRs in cells following cDNAs injection opened new ways to study these receptors in isolation. Furthermore, the recent linkage analysis between a form of autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) with a mutation in the gene coding for the alpha 4 subunit of the neuronal nAChRs constituted the first demonstration that alteration of these receptors may be at the origin of epileptic discharges. Physiological and pharmacological studies of these mutated receptors revealed that the two mutations so far identified in ADNFLE patient cause a loss of function. In this work we shall review, in the light of the latest findings, properties of control and mutated receptors and evaluate how their alteration can be at the origin of nocturnal seizures. PMID- 10472660 TI - [Benign familial neonatal convulsions: a model of idiopathic epilepsy]. AB - Benign neonatal familial convulsions have been recognized as a distinctive epileptic syndrome since 1964. This rare epileptic syndrome was classified in the category of idiopathic generalized epilepsies. Recently, mutations of potassium channel genes (KCNQ2, KCNQ3) were identified as responsible for this autosomic dominant epileptic syndrome. Generalized tonico-clonic seizures start at the second or third day after birth in children with no prenatal or perinatal pathological history. Interictal EEG is normal. This epilepsy is age-dependent: less than ten percent of children present seizures later in life. Despite their rarity, BNFC represent a useful model to understand the pathophysiology of idiopathic age dependant epilepsies. PMID- 10472661 TI - Interictal-ictal interactions and limbic seizure generation. AB - Interictal discharges are used in clinical practice to localize the epileptogenic focus in patients with partial epilepsy. However, the interaction between interictal and ictal discharges remains debatable. For instance, interictal events may lead to seizure onset in some models of epileptiform discharge. By contrast, in other models, disappearance of interictal activity (for example by activation of GABAB receptors) induces or potentiates ictal events. We have recently obtained new evidence for a control exerted by interictal discharges on ictal activity in rodent combined slices of hippocampus-entorhinal cortex. In this preparation continuous application of 4-aminopyridine induces: (i) interictal activity which initiates in CA3 and propagates via CA1 and subiculum to the entorhinal cortex, and return to the hippocampus through the dentate gyrus; and (ii) ictal discharges, which originate in the entorhinal cortex and propagate via the dentate gyrus to the hippocampus. Ictal discharges disappear over time, while synchronous interictal discharges continue to occur. Lesioning the Schaffer collaterals abolishes interictal discharges in CA1, entorhinal cortex and dentate gyrus and discloses entorhinal ictal discharges that propagate, via the dentate gyrus, to the CA3 subfield. Interictal activity of CA3 origin also prevents the occurrence of ictal events recorded in the entorhinal cortex in Mg(2+)-free medium. Moreover, in both models, ictal discharge generation in the entorhinal cortex after Schaffer collateral cut is prevented by mimicking CA3 activity through rhythmic electrical stimulation of CA1 hippocampal outputs. Hence, our data demonstrate that hippocampus interictal discharges control the expression of electrographic seizures in entorhinal cortex. Sectioning the Schaffer collaterals may model the epileptic condition in which cell damage in the CA3 subfield results in loss of CA3 control over the entorhinal cortex. Hence, the functional integrity of hippocampal CA3 neurons may represent a critical control point in temporal lobe epilepsy. PMID- 10472662 TI - [Modification of ictal cerebral blood flow studied by positron emission tomography (PET) and stereoelectroencephalography (SEEG)]. AB - Converging evidence suggests that partial epileptic seizures can cause a circumscribed increase of CBF. However, quantified studies of ictal perfusion changes verified by intracranial EEG are still needed to better evaluate the relationships between CBF changes and the underlying electrical activity. We studied, in 10 patients undergoing a SEEG for epilepsy surgery, the local perfusion changes measured by [15O]-H2O PET during focal epileptic discharges induced by intracerebral electrical stimulation. CBF measurements under SEEG control were performed at rest and during stimulations eliciting, or not, subclinical or mild symptomatic epileptic discharges. PET and magnetic resonance imaging slices parallel to the AC-PC plane were matched for anatomical identification of cortical regions. Individual normalized CBF difference images between rest and stimulation (with or without induced discharge) were analysed by a hierchical description and multiscale detection method. Only CBF changes at p < 0.01 were considered significant. No significant CBF change was observed on test retest at rest or during ineffective stimulations. Among the 12 elicited discharges, of which 11 involved mainly the temporal lobe, 8 were associated with a focal CBF increase (16-55 p. 100), there was no CBF change in 3, and a significant CBF decrease was observed in 1 (64 p. 100). These latter 4 discharges were restricted to mesio-temporal lobe structures. PET data were analysed in the same 10 anatomical areas during each of the 12 discharges. In 70.5 p. 100 of the 61 regions explored by SEEG, PET and depth EEG findings were consistent, showing either a CBF change in a discharging area, or no CBF change in a region unaffected by the discharge. Areas of increased CBF indicated an underlying epileptic discharge in almost 100 p. 100 of the cases. Discrepancies between SEEG and PET concerned almost always discharging regions showing no ictal CBF changes. A focal CBF increase detected at the seizure onset is a reliable marker of an underlying epileptic discharge. However, the extent of the CBF changes can be more restricted than that of the electrical discharge, and ictal discharges restricted to the mesial temporal cortex can be overlooked by CBF studies. PMID- 10472663 TI - [Ictal SPECT in the epileptic child. Contribution of subtraction interictal images and superposition of with MRI]. AB - Ictal SPECT is a highly sensitive method to localize the epileptogenic focus in refractory temporal lobe epilepsy in adults. In extratemporal epilepsy, sensitivity can be improved by subtracting interictal from ictal images and superimposing subtraction images on MRI. In children, such a procedure is potentially interesting because most epilepsies are extratemporal and ictal SPECT not yet routinely developed. The aim of this study was to test the feasibility of ictal SPECT with subtraction image processing in a pediatric population. Twenty six children with refractory partial epilepsy and aged from 3 months to 18 years underwent ictal ECD-SPECT (20 mCi/1.73 m2) combined with video-EEG and interictal ECD-SPECT plus 3D-MRI two days later. Ictal-interictal subtraction images were computed by registering and normalizing the ictal to the interictal SPECT scans for each child. The ictal, interictal SPECT and subtraction images were registered to the children's MRI. Difference images were then superimposed to MRI for anatomical localization of the perfusion changes (overlay images). Looking for perfusion changes, overlay images allowed to detect at least one hyperperfused focus in 92 p. 100 of the 26 children compared to 73 p. 100 visually comparing ictal and interictal scans separately. Seizure onset was suspected on clinical and/or EEG and/or MRI in 19 children. Positive overlay images were concordant (n = 11) or larger (n = 7) than the suspected focus in 17/19 (90 p. 100), whereas they failed to show any abnormality in 1 child and were discordant with MRI in another one. In the 7 remaining patients, images showed cortical localization in 6 cases. Ictal SPECT is therefore faisable in very young children. Ictal-interictal subtraction SPECT images co-registered to MRI improves sensitivity compared to classical visual analysis. It seems therefore to be a helpful technique to localize the onset of seizure and to guide the intracranial recording in childhood epilepsy. PMID- 10472664 TI - Positron emission tomography receptor studies in epilepsy. AB - Investigations with specific PET ligands that bind to specific neuro-receptors give information on abnormalities of neurotransmission involved in the pathophysiology of the epilepsies. Data need to be interpreted in the light of optimal structural imaging. Objective voxel-based and region-based analyses, with correction of partial volume effect, are complementary. Central benzodiazepine (cBZR) and opioid receptors have been studied most. Reduced cBZR binding is commonly seen at an epileptic focus, in a more restricted distribution than an area of hypometabolism, and sometimes also in projection areas. In contrast to acquired lesions causing partial seizures, focal increases in cBZR binding have been demonstrated in malformations of cortical development and also in areas of brain that appear normal on MRI, indicating the widespread nature of the abnormalities. Focal abnormalities of cBZR are also commonly found in patients with partial seizures and normal MRI. It is not yet clear how useful these data will prove to be in presurgical evaluation. Mu- and delta-opioid receptors have been found to be increased in temporal neocortex overlying mesial temporal epileptic foci, but with different patterns of increase. Dynamic studies of the binding of 11C-diprenorphine to opioid receptors are possible using PET, and have implied the release of cerebral endogenous opioids at the time of serial absences and reflex seizures induced by reading. Other tracers, that have been applied less widely, label the enzyme monoamine oxidase type B and peripheral benzodiazepine and histamine H1 receptors. PMID- 10472665 TI - [Interactions between the epileptic network and brain function: an approach by nonlinear analysis of intracranial EEG]. AB - Recent advances in the non-linear dynamics analysis have made it possible to identify hidden recurrences in EEG signals that could be missed by more traditional linear techniques such as power spectrum or coherence analysis. This is particularly true for epileptic EEG recordings either in animals or in humans as epileptic phenomena are usually concomitant with the emergence a strong non linear EEG behavior. Non-linear dynamical analysis techniques quantify the relations between EEG signals. The literature concerning the spatio-temporal characteristics of the epileptic processes during seizures and interictal periods is reviewed. Our attention has been mainly focused on the interdependences between brain structures or on the dynamical changes of one particular brain region during intracranial recordings. These data could explain in part the dysfunctioning of the cerebral cortex induced by epileptic activities and provide an insight into the spatio-temporal organization of the epileptic network. Futhermore, by tracking the time variation of non-linear indices, one can anticipate the occurrence of seizures in temporal lobe epilepsies. All this information could contribute to improve definitions of the epileptogenic zone in partial epilepsy and also open the way to preventive interventions. PMID- 10472666 TI - Magnetic resonance spectroscopy in focal epilepsy: 31P and 1H spectroscopy. AB - Magnetic resonance spectroscopy (MRS) is rapidly becoming a clinical and research tool in epilepsy. Animal studies have demonstrated metabolic abnormalities in the interictal and ictal state showing energy depletion and changes in neuronal compounds. Similarly, clinical studies have demonstrated consistent abnormalities involving energy (31P) and cellular dependent (1H) compounds. Phosphorus MRS (31P) shows lateralizing metabolic dysfunction in approximately 65-75 p. 100 of patients with temporal lobe epilepsy (PCr/Pi). Proton MRS (1H) using single-voxel or chemical shift imaging has demonstrated a high sensitivity in lateralizing temporal lobe epilepsy (65-96 p. 100) with bilateral changes seen in 35-45 p. 100 of patients. The role of MRS in extra-temporal lobe epilepsy is less accurate because of the spatial limitations of current techniques. Further advances in this field promise to improve the clinical utility of MRS in epilepsy. PMID- 10472667 TI - Source localization in refractory partial epilepsy. AB - In this paper, 51 patients with refractory complex partial seizures (CPS) and intracranial structural abnormalities demonstrated with optimum MR (space occupying: n = 16; atrophic: n = 32; dysplastic: n = 3) were studied. Video-EEG monitoring showed CPS in all patients. In 13 patients, additional intracranial EEG monitoring demonstrated hippocampal seizure onset in 12 and medial occipital ictal onset in 1 patient. Interictal and ictal dipole modeling using a spherical head model and realistic electrode coordinates were performed. Spatiotemporal dipole mapping of interictal epileptic discharges revealed two distinct dipole patterns. Patients with lesions located in the medial temporal lobe (n = 41) and medial occipital lobe (n = 2) uniformly presented a dipole with an elevation of more than 15 degrees relative to the axial plane. Eight out of ten patients with extratemporal lesions and 1 patient with a pure neocortical temporal lesion had a less stable dipole with an elevation less than 15 degrees relative to the axial plane. Dipole modeling of epochs of early ictal discharges revealed a striking correspondence with the interictal findings in individual patients. Ictal dipole modeling identified the ictal onset zone correctly when compared with intracranial EEG recordings from bilateral hippocampal depth electrodes in patients with medial temporal seizure onset. Mapping of dipoles on MR images of individual patients facilitated clinical interpretation of the EEG data. Interictal and ictal dipole mapping provided additional and clinically relevant information and may obviate the need for intracranial EEG studies in some surgical candidates for refractory CPS. PMID- 10472668 TI - [New methods for evaluation of memory in temporal lobe epilepsy: a functional imaging approach]. AB - Temporal lobe epilepsy (TLE) is usually associated with a specific impairment of memory functions. A reliable assessment of memory function seems therefore a crucial element of the presurgical evaluation for surgery epilepsy, especially for temporal lobectomy. To date, intracarotid amobarbital testing remains the gold standard to evaluate memory before planning surgery in patients with refractory TLE. Functional imaging, either positron emission tomography or functional MRI, holds great promise as a powerful tool in memory evaluation. Preliminary studies, essentially in healthy volunteers, have shown promising results. In the future, functional imaging, combined with other techniques, will certainly contribute to the clinical presurgical evaluation of patients with epilepsy. PMID- 10472669 TI - Introduction. The economics of animal disease control. PMID- 10472670 TI - The application of economics in animal health programmes: a practical guide. AB - Economic analysis is a valuable technique in the planning and management of animal health programmes at the national policy level and at the level of individual livestock enterprises. Considerable experience has now been accumulated in the adaptation of methods of economics to address decisions on disease control issues. Different methods are appropriate for endemic diseases, for diseases which occur intermittently as local outbreaks, and for diseases which have the potential to cause an epidemic. A guide to the selection of the most suitable method is provided. The information required for analysis at individual herd level is much simpler than that required for national policy decisions and funding decisions by international agencies; an outline of the requirements for each level of analysis is provided. Issues surrounding the degree of risk of different choices are examined, and the application of decision analysis to higher-risk choices is introduced. Ways in which difficulties and hazards can be overcome are suggested for those unfamiliar with the techniques. Finally, procedures to provide confidence in the results of evaluations through sensitivity analysis are explained. PMID- 10472671 TI - Methods of economic impact assessment. AB - A broad framework for the assessment of the economic impact of animal diseases and control of these diseases is described. Various levels of analysis are discussed, from the herd, to the household, the sector, the nation and the region. For each of these levels, methods of economic impact assessment are outlined and discussed with respect to the objectives, data requirements and outputs of each type of assessment. In addition, the authors attempt to describe the conditions under which the various methods might be used. To conclude, a discussion of the state of the art is presented, and areas of further research are identified. PMID- 10472672 TI - The economic implications of animal diseases and disease control at the national level. AB - The authors examine the economic implications of animal diseases and control programmes at the national level, including the role of government in animal health, the effect of regulations and the use of cost-benefit analysis. Special attention is paid to the role of economic analysis in government decision-making processes. Economics provides a framework for gathering information and for the presentation of that information in a methodical manner, thereby providing a method for the decision maker to examine policy alternatives. In addition, assumptions underlying the analysis must be clearly laid out and explained by the person undertaking the analysis. Economic reasons for government intervention in animal health programmes include externalities, natural monopolies, public goods, coordination failure, information failure and distribution issues. An integrated holistic approach that includes national and international policy objectives is outlined in the paper. In the approach outlined, government coordinates the activities of stakeholders in animal health, including producers, consumers and researchers. PMID- 10472673 TI - The economics of animal health in farmed livestock at the herd level. AB - Animal health economics is a relatively new discipline which makes use of concepts, procedures and data to support the decision-making process with the objective of optimising animal health management. As the economic impact of most diseases that afflict farmed livestock is typically greater at the sub-clinical rather than the clinical level, animal health management often involves decisions regarding expenditure on preventive measure as well as the treatment of obviously sick animals. Animal health programmes have been shown to provide a very high return on investment. This is because reduction of the impact of disease increases the efficiency of production, often without the need for additional inputs such as feed or labour. The analytical techniques described in this chapter are the simplest, most useful, and most commonly used by animal health economists. Most decisions in animal health economics at the farm level can be arrived at by using partial budgeting, decision tree analysis, or cost-benefit analysis techniques--either alone or in combination. Regardless of the technique chosen, the analysis is only as good as the quality of the data used. PMID- 10472675 TI - The economic impacts of endemic diseases and disease control programmes. AB - The authors discuss the evaluation of the economic impacts of endemic livestock diseases, and economic issues in control of these diseases. Particular attention is focused on helminths and on endemic vector-transmitted infections (particularly ticks and tick-borne diseases). Decisions relating to disease control have to be made by government and by the producer. Government requires information on the level of control to adopt, the extent of involvement needed, and how to fund animal health programmes (particularly how to share costs between taxpayers and livestock producers). Individual producers require information as to how much effort to invest in disease control, including information collection effort, and how to design control strategies. Economics can shed light on these issues. However, experience suggests that animal health policies are particularly difficult to evaluate from an economic viewpoint, with complex relationships between animal health, production impacts, market access, and non-production benefits of livestock. While little information is available concerning the cost of helminth diseases, many estimates have been made of the costs of ticks and tick-borne diseases at a regional and national level, sometimes demonstrating that eradication is warranted. PMID- 10472674 TI - The economic evaluation of control and eradication of epidemic livestock diseases. AB - Many countries have implemented strategies to control and eradicate epidemic diseases. These strategies are usually based on either stamping-out or routine vaccination, sometimes complemented by emergency vaccination. The authors describe these strategies, using examples to illustrate each one. The economic evaluation of control and eradication of epidemic diseases is a complex matter. The authors provide further insight into this area by describing the various elements involved in both the 'non-outbreak periods' and the 'outbreak periods'. In addition, a system of categorisation of the direct costs and consequential losses is suggested for the calculation of costs and losses incurred by outbreaks. The economic impact of epidemic diseases on farmers and the livestock sector as a whole differs; these differences may be influenced by the control and eradication strategies applied. An attempt is made to provide a basic framework for economic evaluation on various economic levels. PMID- 10472676 TI - The economics of optimal health and productivity in smallholder livestock systems in developing countries. AB - Livestock kept or produced in smallholder farming systems are an important component of the agricultural economy in the developing world. The role of livestock on smallholder farms varies widely, providing draught power for crop production or as a production activity for subsistence needs or market sale under systems ranging from extensive pastoralist to intensive, peri-urban feeder and dairy systems. A set of unique conditions and features characterise smallholder systems, and these need to be appreciated when assessing the strategies that have evolved for managing animal health in smallholder systems, and evaluating opportunities for improving disease control strategies. To provide a framework for discussing animal health issues and analytical methodogies, a typology of smallholder livestock and crop/livestock systems is developed. The typology considers livestock systems both in terms of the degree of intensification, as measured by market orientation and intensity of factor use, and in terms of importance within the household economy, as measured by contribution to household income. A number of characteristics are identified that distinguish smallholder systems from the commercialised systems of developed countries, including the multiple functions livestock serve, the integrated nature of livestock activities, multiple objectives of producers and lower capacity to bear risk at the household level, as well as poor infrastructure, markets, and access to information at the community level. Three representative smallholder livestock systems from Africa are described in detail, highlighting the relevant characteristics and the implications for analysing disease control strategies. Smallholder dairy systems in Kenya demonstrate the role of individual producer decision-making for animal health management in intensive, market-oriented systems, placing emphasis on farm-level risk and production management aspects of disease control. In extensive pastoralist systems where epidemic disease are still important and infrastructure is poor, disease control primarily involves managing communal natural resources, requiring a different analytical approach. Finally, in crop farming systems using draught cattle, the livestock activity is an integrated component of crop production and this must be reflected in the approach used to evaluate draught animal health management. Continued development of analytical approaches and decision-support tools for disease control strategies adapted to the special characteristics of these systems will be needed as smallholder systems continue to intensify in areas with good market access, and those in marginal areas face increasing pressures to optimally manage the natural resource base. PMID- 10472677 TI - The economics of the delivery of veterinary services. AB - Low livestock productivity in many developing countries is commonly considered to reflect, among other factors, the inadequate supply of services to control disease. Veterinary services have traditionally been provided by the state, but public finance constraints have limited the availability and effectiveness of public services. The author explains how economic theory can be used to identify alternative delivery systems (beyond the state) for providing animal health care and proposes new roles for the state and private sector in service delivery. The author highlights a number of barriers that currently limit the potential contribution of the private sector to service delivery, and describes a variety of approaches that have been used by the state to create an enabling environment for the private sector. PMID- 10472678 TI - The economic implications of greater global trade in livestock and livestock products. AB - The Uruguay Round of the General Agreement on Tariffs and Trade (GATT) established the World Trade Organization to supervise the reduction of barriers to, and liberalisation of, world trade. The application of sanitary and phytosanitary measures will be standardised to avoid use for protectionist purposes by countries or regional trade blocks. Harmonisation of animal disease control measures within regional blocks is essential if benefits to freer trade are to occur, but this harmonisation must be balanced against potential disease risks and costs associated with disease outbreaks. World trade in livestock products is concentrated among developed countries, although developing countries are responsible for approximately a third of poultry meat imports and exports. Despite liberalisation, the share of global trade by developing countries is unlikely to increase greatly in the short term. The benefits of trade and of freer trade are emphasised. Examples are given of the impacts of trade barriers on developing countries and of the harmonisation of European Union animal health standards. Economic implications for the future of greater global trade are assessed. PMID- 10472679 TI - Economic impact assessment of rinderpest control in Africa. AB - The authors assess the economic impact of the Pan-African Rinderpest Campaign (PARC). The PARC programme commenced in 1986 with the objective to control and ultimately eradicate rinderpest from Africa. From among the thirty-five countries that participated in PARC, ten countries were selected for the analysis, based on data availability. The three following key socio-economic issues were addressed: cost-effectiveness, returns to investment and the welfare gains of the intervention. The standard cost-benefit approach based on a computer spreadsheet model was used to assess the economic impact of rinderpest control. Benefits of the intervention consisted of increased revenue due to avoided production losses. Estimates of the value of production losses were obtained under both 'with PARC' and 'without PARC' scenarios and the incremental benefits were derived as the difference between the two scenarios. In addition, an economic surplus model was used to assess the distribution of welfare effects generated by the intervention. Analysis of funding for the national campaigns showed roughly equal commitment to the programme by national governments and the principal donor, the European Union. Examination of the implementation costs in the ten countries indicated that with the exception of one country, PARC was implemented in a cost-effective manner with average costs appearing within a relatively narrow range. The figures obtained in ECU (European currency units) were between ECU0.27 and ECU0.60 per head of cattle vaccinated. The estimated average return from the ten countries (ECU1.8 for each ECU invested in the campaign) demonstrates that based on the sample of countries, rinderpest control in Africa has been economically profitable. In each of the ten countries, estimated benefits at least covered the value of the investment in PARC. The programme has provided a total net present value of ECU29 million for the ten countries, suggesting that the implementation of PARC has been a wise public investment decision. Analysis of the distribution of the welfare gains from PARC revealed that producers derived the greater share of the ECU58 million in net value of production losses avoided due to rinderpest control in the ten countries. Consumer gains accounted for approximately one fifth of the total, due to lower prices from increased supplies. PMID- 10472680 TI - The economic impact of foot and mouth disease and its control in South-East Asia: a preliminary assessment with special reference to Thailand. AB - A pilot study of the economic impact of foot and mouth disease (FMD) in the countries and region of South-East Asia is described. Previous economic impact assessments are reviewed and summarised and a synthesis of these contributions is constructed. A framework for the future economic impact of the disease is then developed, incorporating analyses at the sectoral (production system), national and regional levels. Data requirements for such studies are also identified. Integrated epidemiological and economic models for impact assessment were developed and applied to the case study country of Thailand. The models were used to evaluate the economic viability of FMD control programmes in the country. Scenarios evaluated include the effect of improving vaccination coverage and thus reducing productivity losses, and the effect of eventual eradication of the disease. The results indicate that economic returns to the high expenditures incurred in FMD control could be achieved in the short term if greater international trade in pork products was made possible and export prices higher than those in the domestic market could be attained. If FMD were to be eradicated from Thailand in 2010, the eradication would be economically viable, even without exports, with a predicted benefit-cost ratio of 3.73. With additional exports, the economic justification for control becomes much stronger with a benefit-cost ratio of up to 15:1 being achieved. If eradication is not achieved until 2020, returns remain positive without exports, but at a lower rate. The authors propose that the integrated epidemiological and economic models developed be applied to other countries of the region to gain a more accurate insight into the future benefits of FMD control and eradication in the region. PMID- 10472681 TI - [Economic impact of animal diseases on production systems in South America: case studies]. AB - The authors discuss the relative importance of the livestock sector in South America, in developing countries and in countries world-wide. The development of animal health services is highlighted and the new challenges which these services must meet, with respect to the restructuring of international trade, are described. The economic impact of animal diseases and of disease control is evaluated by taking a regional approach and by analysing production systems. A classification of production systems is accompanied by case studies on the economics of animal health. Emphasis is placed on national control programmes, diseases which affect reproduction, parasitic diseases and mastitis, with particular reference to the livestock sector of Colombia. The authors suggest that integrated livestock development programmes include the management of animal health and production information. A change of attitude is also recommended with greater participation of those involved. The need to co-ordinate sustained research initiatives is stressed. PMID- 10472682 TI - The economics of optimal health and productivity in the commercial dairy. AB - Dairy production practices are changing; in order to remain viable, producers must optimise the health and productivity of dairy herds in economic terms. Health care is important in economic terms because disease can substantially reduce the productivity of individual animals. Preventive disease control programmes can thus result in economic gains for the dairy producer. The author describes new approaches to preventing postpartum diseases and dealing with fertility problems which can result from these diseases. Other aspects of dairy production are also changing, employing new technologies where these are judged to be profitable. Innovations include: the use of bovine somatotropin; systematic breeding/culling programmes; new mathematical modelling techniques to determine optimum feed composition and to define optimal growth levels for accelerated heifer-rearing programmes; the use of computers to collect, store and analyse data on animal production and health; and semen selection programmes. Increasing awareness of bio-security is also vital, not least because of the large investment present in dairy herds. Whatever practices are employed, they must offer economic returns to producers that compete with alternative uses of capital. Optimal levels of disease control must be determined for a particular production situation, taking into account not only the economic health of the producer, but also the well-being of the animals. PMID- 10472683 TI - [Role of economic studies in animal health decisions: Example of the cost-benefit ratio of eradication of bovine viral diarrhea in France]. AB - To help livestock production groups to rationalise health decisions, and at the request of the Association for the certification of livestock health (Association pour la certification de la sante animale en elevage: ACERSA), an economic study was conducted to assess the possible cost-effectiveness of the eradication of bovine virus diarrhoea in France. The study was performed using a fictitious average region comprising 235,000 cattle belonging to 3,300 farms, which corresponds to one-eighty-fifth of the total cattle population of France. In the first phase of the study, the cost of the disease in this region was estimated to be approximately six million French francs (US$989,937) per year. Subsequently, the cost of an eradication strategy based on the inspection of all animals when introduced into a herd, the screening of permanently-infected immunotolerant animals (IPI) and the elimination of these animals, was evaluated at nearly eleven million francs (US$1,814,884) during the first year. Theories were then formulated regarding the time required to achieve eradication (twenty years) and to reduce the epidemiological parameters (development curve of the eradication of IPI animals and of animals which had given positive results to serological tests). The reduction in the cost of the disease as a result of the eradication policy was then simulated in accordance with the evolution of the epidemiological parameters. Finally, the cost of controlling the disease, together with the residual cost of the disease, were compared with the cost of the disease without control measures. This demonstrated that such an eradication policy would, in theory, only begin to become cost-effective after approximately fifteen years. In view of the long period required to achieve cost-effectiveness, the considerable complexity of implementing an eradication programme and imponderables (particularly concerning virus spread), the recommendation of such a course of action to cattle-breeder groups is questionable. This research demonstrates that a relatively simple economic approach can serve as a useful decision-making aid. PMID- 10472684 TI - Costs and benefits of rabies control in wildlife in France. AB - The author presents an evaluation of the cost of wildlife rabies in France. This study included the vaccination of domestic animals, the reinforcement of epidemiological surveillance networks and the support provided to diagnostic laboratories, the expenses associated with outbreaks of rabies (animal losses and associated economic losses), the clinical observation of those animals which had bitten humans and the preventive vaccination and post-exposure treatment of humans. A substantial percentage (72%) of this cost was the preventive vaccination of domestic animals. In France, as in other European countries in which the red fox (Vulpes vulpes) is the species most affected, to main strategies for controlling the disease at the reservoir level were evaluated, namely: fox depopulation and the oral vaccination of foxes. The combined costs and benefits of rabies and of both strategies were compared and included either the cost of fox culling or the cost of oral vaccination (baits, bait delivery and follow-up to ensure the efficiency of the vaccination). The cumulative annual costs of both strategies remained comparable until the fourth year, after which the oral vaccination strategy became beneficial. This forecast was made in 1988, readjusted in 1993 and confirmed by ex post analysis five years later. The expected benefits of oral vaccination have now been obtained. Fox depopulation has only ever resulted in a transient lull in the occurrence of the disease, while oral vaccination has proved to be capable of eliminating rabies even in situations in which fox populations were increasing. PMID- 10472685 TI - The new institutional economics of privatising veterinary services in Africa. AB - The authors review a number of critical issues in the structural reform of animal health services for both small and non-commercial livestock producers in Africa and highlight several problems that others concerned with the privatisation of this service area have tended to neglect. Most notably, attention is called to the following: a) the need to retain a central role for paraprofessionals in the new delivery system b) the important and problematic relationship between the veterinary and paraveterinary professions c) the importance of developing state contracting procedures for assisting the private delivery of animal health services that will avoid the problems of local monopoly d) the central role that professionalism will have to play in this area, if collective goods and the public interest are to be served. PMID- 10472686 TI - [Application of the gel test using and anti-IgA antiglobulin for the immunologic diagnosis of autoimmune hemolytic anemia with a negative direct Coombs test]. AB - Autoimmune hemolytic anemias (AIHA) are characterized by hyperhemolysis associated with the presence of the immunoglobulins IgG, IgM or IgA on the red cell membrane. These immunoglobulins react as auto-antibodies against the red cell auto-antigens of the patient. The diagnosis is supported by clinical and biological signs of hemolysis, and by the identification of the auto-antibodies using the direct antiglobulin test (DAT). Here we report 14 cases of patients who showed the clinical and biological profile of AIHA, but who gave a negative DAT. We therefore tried to determine the presence of IgA on the red cell membrane with a method more sensitive than the DAT: the gel test using anti-IgA. With such a gel test, we demonstrated that there were IgA auto-antibodies on the red cell membrane in the 14 cases, therefore confirming the diagnosis of AIHA. We discuss the interest of performing a gel test with anti-IgA in each case where AIHA is suspected, but in which a negative DAT has been observed. PMID- 10472687 TI - [Risk behavior among blood donors: efficacy of a new questionnaire]. AB - The clinical selection of volunteers for blood donation is essential to reduce the risk of viral transmission by blood transfusion. The aim of this study was to evaluate a new questionnaire for a pre-donation medical interview. This questionnaire was developed by transfusion practitioners, epidemiologists and professional investigators, and focused on risk behaviors of blood donors and their partners. Five blood banks in the French Ile-de-France region (around Paris), participated in the study from May 1995 to January 1996. All participating doctors were specifically trained by professional investigators. The sex and the age of donors, the type of collection, the duration of interviews and the reasons for exclusion from donation were recorded. The results were compared to those of a prior study dealing with a sample of 15,000 donors presenting the same characteristics, whose blood was taken of at the same collection sites in 1993. Of the 1,527 volunteers donating blood, 14% were interviewed in fixed centers and 86% by moving teams (38% in firms, 22% in towns, 13% in civil service facilities, 13% in school or academic centers). For 15.9% of the volunteers, this was the first donation (range: 7.3% in fixed centers to 41.5% in school and academic centers). The mean duration of the interview was 11 min (10 min for volunteers included, 14 min for donors excluded from donation). It decreased from 14 min at the beginning of the study to 10 min by the end of the study. The percentage of donors excluded for risk behavior (3.7% in 1995-96 vs 1.5% in 1993, P < 0.001), or medical reasons (12.2% in 1995-96 vs 8.4%, in 1993, P < 0.001) was significantly greater in 1995-96 than in 1993 (15.9% vs 9.9%, P < 0.001). In 1995-96, 35.0% of exclusions for risk behavior were related to male homosexuality, multiple partners or the risk behavior of the partner vs 12% in 1993 (P < 0.001). The risk of exclusion was 5.5 times higher for donors not living in a couple. The results obtained demonstrated the value of this new approach to the medical interview. PMID- 10472689 TI - [The new International Standardization Organization 9000 norms: 2000]. AB - The reviewing of ISO 9000 norms has been planned to be published within the last quarter of the year 2000. It aims at overcoming the limits and difficulties of use evidenced by the implementation of the 1994 ISO norms, and at providing an analysis aid for improving quality, which would be adapted to the needs of each company or user. What is at stake with these new norms is the implementation of a quality management system, and its evolution towards a total-quality management. PMID- 10472688 TI - [DNA polymorphism applied to paternity testing. Analysis of 877 cases]. AB - Paternity testing is based on biological analyses that have drastically developed during the past 20 years. According to scientific developments, paternity testing was based on red blood groups studies, the analysis of red cell enzymes and plasma proteins polymorphisms, the typing of the HLA-A and B antigens, and the DNA polymorphism in its various forms. This study aims at comparing the various analyses so that each becomes a reference for the others, in order to determine (i) the acceptable level of the paternity probability evaluation, and (ii) the exclusion rules according to new technologies. The relative performances of the various combinations of tests is analyzed in this report, based on a study of 877 cases. Thanks to studies based on the gene amplification of micro-satellites, the efficiency of this technique has been proved: it is however necessary to identify the limits of the tests and to know how to deal with those cases which require further analysis. Beyond the most efficient biological analysis, it is very important to think about paternity testing as a process in which biological tests are only one step. PMID- 10472690 TI - [Pharmacovigilance in practice]. AB - Directed by the French Agency for the Safety of Health Products (AFSSAPS), the French pharmacovigilance system is in charge of the surveillance of drugs after they have been provided by AFSSAPS with official marketing authorizations that are in France either 'new drug approval certificates' (AMM) or 'temporary utilization authorizations' (ATU). About 3,700 pharmaceutical products are concerned which are used either for treatment (all drugs and remedies, inclusive plasma-derived blood products), prevention (vaccines, oral contraception), diagnosis (contrast products, ...), or to modify a physiologic function (general or local anesthetics). At the national level, the main actors of the system are AFSSAPS and its National Commission, the 31 Regional Centers of Pharmacovigilance, all the health professionnals, and the pharmaceutical laboratories. Health professionnals are held to notify any suspected serious or unexpected adverse effects as quickly as possible. The analysis of data collected by the national report bank permits alerts and inquiries about drug safety. Furthermore regional centers of pharmacovigilance are responsible for drug information. The French pharmacovigilance system works in cooperation with the European Agency for the Evaluation of Medicinal Products. PMID- 10472691 TI - [The national blood donation promotion campaign]. PMID- 10472692 TI - [Training for health care facility nursing personnel about transfusion safety: use of visual aides]. PMID- 10472693 TI - [Clinical research today and its promotion by industry--advantages and risks]. PMID- 10472694 TI - [Partnership between drug industry and the university--innovation, proof of efficacy, drug safety]. PMID- 10472695 TI - [Promoting research and graduate education by the pharmaceutical industry- scientific progress or marketing strategy?]. PMID- 10472696 TI - [Does financing researchers and research at university clinics entail a conflict of interests and dependence?]. PMID- 10472697 TI - [Legal aspects of support to clinics, research and continuing education by industry]. PMID- 10472698 TI - [Preemptive effects caused by co-analgesia with ketamine in gynecological laparotomies?]. AB - The preemptive use of analgetics makes it possible to influence sensitization proceedings caused by a trauma. Various mechanisms are effective in central pain treatment. The NMDA receptor plays an important role. The investigation presented was to examine whether an intraoperative combination of analgetics with different points of contact leads to improved postoperative analgesia. Altogether 39 female patients who had to undergo a gynaecological laparotomy were examined. Three groups of 13 patients were randomly formed. In addition to usual general anaesthesia, the patients of group one received 0.5 mg/kg bodyweight ketamine racemate before the skin incision and thereafter 10 micrograms/kg/min ketamine infusion continuously until peritoneum closure and then sodium chloride 0.9% as a placebo after the final skin suture. The patients of group two received placebos before the skin incision and intraoperatively and 0.5 mg/kg bodyweight ketamine after the last skin suture. In group three the patients received placebos at all three points of time. Analgetics consumption, pain intensity, awakening reaction, vital parameters as well as psychomimetic side-effects and nausea/vomiting were listed postoperatively. Between the groups no differences were found regarding postoperative analgetics consumption. In addition, the pain intensity showed no differences regarding an improved postoperative analgesia through the combination of analgetics with different points of contact. The intraoperative combination of ketamine and alfentanil does not lead to a reduction of postoperative pain. No preemptive analgesia is clinically provable. PMID- 10472699 TI - [Preoperative clonidine comedication within the scope of balanced inhalation anesthesia with sevoflurane in oral surgery procedures]. AB - Both clonidine and sevoflurane are interesting drugs for anaesthesia in maxillo facial surgery. The present study was performed to discover how far it is possible to combine the benefits of sevoflurane (fast modulation of depth of anaesthesia, rapid emergence and recovery) and clonidine (reduction of perioperative stress response, prophylaxis of postoperative shivering, analgetic, antiemetic and anaesthetic-saving effect) without compromising the pharmacokinetic of sevoflurane. Twenty-eight patients were included in the present double-blinded prospective study. These patients were randomly treated with an infusion of 4 micrograms kg-1 clonidine (group 1) or a placebo (group 2) preoperatively. For anaesthesia a standardized procedure with fentanyl, propofol, rocuronium, N2O/O2/sevoflurane and an antiemetic prophylaxis with DHB was performed. The depth of anaesthesia was controlled by using spectral edge frequency (target--SEF90 = 10 Hz). Perioperative stress response was assessed by noting the effects on haemodynamic parameters (MAP, heart rate), and emergence and recovery were assessed by using established standardized tests. We confirmed the anaesthetic-saving property of clonidine only for fentanyl (-20%). On the other hand, there was no difference in MAC-sevoflurane values between the groups in keeping a steady target--SEF90 (1.62 +/- 0.26 versus 1.65 +/- 0.24 vol.%). The time until emergence and recovery was not significantly different. Even the occurrence of PONV, the VAS level or the postoperative analgesic requirement did not differ in the two groups. However, the incidence of postoperative shivering was significantly higher in the placebo group. The stress response to intubation or extubation was lower in the clonidine group. The haemodynamic parameters in the clonidine group were intraoperatively always below the baseline, in some cases by more than 20%, making therapy for hypotension or bradycardia frequently necessary. Postoperatively, the majority of the patients showed similar changes in these parameters, but did not reach the 20% mark. Preoperative clonidine comedication seems to complicate the management of anaesthesia. On the other hand, it is beneficial during the early postoperative period (e.g. stability in haemodynamics, prophylaxis of shivering) without compromising emergence and recovery. Our results show that therapy with clonidine should be better placed at the end of anaesthesia. PMID- 10472700 TI - [Portable capnographs in emergency care: a comparison of equipment]. AB - Recently, transportable capnographs fulfilling the practical demands of emergency medicine have become commercially available (NPB-75, sidestream, Nellcor Puritan Bennett; and Tidal Wave, mainstream, Novametrix). A prerequisite for their use is an accuracy as required for clinical purposes (i.e., pCO2 +/- 2 mmHg). Additionally, environmental conditions in emergency medicine (e.g., changes in ambient temperature) should not have a significant impact on accuracy. The objective of this investigation was to analyse the accuracy of the two capnographs. The accuracy of the pCO2 measurement was evaluated under the following conditions: (1) measurement with three gas mixtures of defined concentrations (gas A: 5% CO2, 95% O2; gas B: 5% CO2, 20% O2, 75% N2; gas C: 10% CO2, 90% N2) related to STPD conditions (STPD = Standard Temperature and Pressure, Dry); and (2) exposure to changes in temperature (from +22 degrees C to -20 degrees C, and from -20 degrees C to +22 degrees C) applying the aforementioned 3 gas mixtures (STPD); and (3) in 20 patients manually ventilated with pure oxygen following endotracheal intubation (i.e., BTPS conditions = body temperature and pressure, saturated). Adequacy of the results was compared to the alveolar gas monitor AGM 1304 (Bruel & Kjaer, Copenhagen, Denmark; sidestream) which served as the reference method (providing an accuracy for the alveolar carbon dioxide partial pressure (pACO2) of +/- 1 mmHg). In the 3 dry gas mixtures, mean inaccuracy proved to be +4.5 +/- 4.1, +2.8 +/- 3.7, and +2.2 +/- 7.0 mmHg (gas A, gas B, gas C; STPD) with the Nellcor sidestream device. Using the Novametrix mainstream capnograph the results were found as follows: (1) -1.1 +/- 0.6, +2.9 +/- 0.6, and +5.6 +/- 2.3 mmHg (oxygen compensation enabled); and (2) +0.2 +/- 1.6, +2.2 +/- 0.6, and +3.2 +/- 4.2 mmHg (oxygen compensation disabled). After changing the environmental temperature (-20 degrees C / +22 degrees C), the resulting deviations (gases A-C, STPD) found with the Nellcor device averaged -12 +/- 4% and +15 +/- 3% (Nellcor); with the Novametrix mainstream device the deviations averaged -1 +/- 2% and +1 +/- 1%, and -2 +/- 1% and +1 +/- 1% (oxygen compensation enabled/disabled). Mean inaccuracy of the pCO2 measurement during ventilation of patients with pure oxygen (BTPS) was found to average -0.9 +/- 0.9 (Nellcor), and either +3.9 +/- 0.8 or +2.1 +/- 0.7 mmHg with the Novametrix (oxygen compensation enabled/disabled). Under BTPS conditions, both devices showed an acceptable deviation of the measurement accuracy up to a maximum of +/- 2 mmHg. The higher deviations of the "NPB-75" (Nellcor Puritan Bennett, sidestream) when using dry gas mixtures (STPD) may be explained by the automatic water vapour correction. Under the conditions of low and changing ambient temperature (-20 degrees C, +22 degrees C), only the "Tidal Wave" (Novametrix; mainstream) remained uninfluenced, whereas deviations of -12% and +15% were found with the "NPB-75". PMID- 10472701 TI - [Value of the laryngeal mask in emergency care--a survey of North German emergency physicians]. AB - In clinical routine, the laryngeal mask airway (LMA) has proved an alternative to both endotracheal intubation and mask ventilation. In a survey among North German emergency physicians, aspects such as doctors' acquaintance with the LMA, the degree of ist distribution, its use and its potential benefits under non-hospital emergency conditions were evaluated. Seventeen per cent (n = 162) of physicians responded to the questionnaire, 75% of them (n = 122) anaesthetists. Although 73% (n = 119) were familiar with the use of the LMA from clinical experience, and again 73% would welcome having LMA use and application as part of the training of non-academic members of ambulance teams, only 24% (n = 37) of the responding doctors have an LMA on their emergency ambulances and merely twelve (7.4%) physicians reported actually using the LMA, one of them unsuccessfully. Sixty three per cent considered the LMA the first-choice alternative in an unexpected "can't-ventilate-can't-intubate" situation. Use of the LMA should be extended in emergency medicine especially as its application is relatively easy to learn in clinical routine. PMID- 10472702 TI - [Apoptosis]. AB - The aim of this review is firstly to summarize what we know at the present time on the structure of the apoptotic cell and on the main biochemical changes which characterize this type of cell death. Emphasis is put on the functions of a new group of proteases, the caspases, which play an essential role in cell demise. Description of the molecular mechanisms involved in apoptosis, is developed in the second part of the review, where the place of the bcl-2 gene superfamily is detailed such as the many results showing that mitochondria could play a key role in apoptosis regulation. Cell integration of these new findings with regard to apoptosis is illustrated by the description of the apoptotic Fas pathway, the mode of action of the proapoptotic cytokine TNF alpha and the analysis of the apoptosis mechanism when DNA is damaged and when the p53 gene is overexpressed. In the third part, the many circumstances where apoptosis occurs is detailed not only in physiological but also in pathophysiological and pathological conditions. The place of apoptosis and its therapeutic consequences in many human diseases are illustrated by some examples. PMID- 10472703 TI - [Apoptosis and cerebral ischemia]. AB - Although physiological cell death has been known for decades, interest in the subject was renewed in 1972 when Kerr, Wyllie and Currie described in detail the ultrastructural changes characteristic of dying cells and coined the term apoptosis to describe the process. When dying during development or following a toxic insult, cells display a wide variety of morphological changes. A binary classification scheme suggests that physiologically appropriate death is due to apoptosis, and that pathological mechanisms involve necrosis. In this report, we will address developments in our understanding of a potential involvement of apoptotic cell death in ischemia which induce selective and delayed neuronal degeneration. Such results may open important steps in therapeutic approaches for the preservation of neurons. PMID- 10472704 TI - [Apoptosis and gamma rays]. AB - Gamma radiation can induce cell death in lymphocytes. Apoptosis is characterized by numerous morphological, biochemical and molecular modifications measurable using various methods. Some radioprotectors have antioxidant properties and are able to inhibit radiation-induced DNA fragmentation and caspase activation. There are several caspases that cleave proteolytically many proteins and trigger phosphatidylserine externalization recognized by phagocytes. Three main proteins are involved in the regulation of radiation-induced apoptosis: p53, Fas and Bcl 2. The pharmacological regulation of cell death is discussed in order to investigate the subsequent effects related to cell regeneration following radiation injury. PMID- 10472705 TI - [Targeting bone marrow with the help of polyalkylcyanoacrylate nanoparticles]. AB - Using a mouse model, we examine drug targeting towards bone marrow. One cytotoxic (doxorubicin) and one stimulating (rhG-CSF), bound to polyalkylcyanoacrylate nanoparticles, were studied. Histological studies, using a fluorescence microscope, showed rapid capture of nanoparticles by bone marrow macrophages and granulocytes as soon as 15 minutes after injection into the blood stream. Doxorubicin nanoparticles, administered at a dose of 11 mg/kg were more toxic than free doxorubicin on all blood and marrow cell lines. Moreover, the choice of the nature of the polymer had an influence on toxicity: doxorubicin polyisohexylcyanoacrylate nanoparticles were more toxic than polyisobutylcyanoacrylate particles. Quantification of doxorubicin in bone marrow has confirmed these results. The bone marrow concentrations observed demonstrated that there was a high level of targeting towards the bone marrow that would be very interesting to use for a stimulating drug. Nevertheless, rhG-CSF nanoparticles did not show better efficacy than free rhG-CSF. PMID- 10472706 TI - [Binding affinity to steroid hormone receptors and antiproliferative action on MCF-7 cells of coumarin derivatives and isoflavonoids]. AB - The study of the relative binding affinity to steroid hormonal receptors and of the antiproliferative action on MCF-7 cells of 3-arylcoumarines, 3-aryl-4 hydroxycoumarines, isoflavanones, isoflavan-4-ols and isoflavones, indicates a weak activity in the case of some representatives of coumarines. PMID- 10472707 TI - [Generic drugs and right of substitution. Relating to Article 29 of the 1999 *social Security Financing Act]. AB - The 1999 Social Security Financing Act grants the right of substitution to pharmacists. If the prescriptor does not object, the pharmacist can replace a medicine by its generic. After reiterating that generic medicines are essentially similar but not strictly identical to the original or to each other, we analyze the discussions which took place at the National Assembly and Senate which should have resulted in some further modifications to the recently adopted legislative text. PMID- 10472708 TI - [Comparative advertising and medicine: changing of regulations]. AB - Directive 97/55/EC of October 6, 1997 has just amended Directive 84/450/EEC concerning misleading advertising in order for it to encompass comparative advertising. The purpose is to harmonize regulations regarding comparative advertising. Although the basic terms of this Directive are very similar to those imposed in France by the law of January 18, 1992, its transposition should, however, require changes to be made to French law. PMID- 10472709 TI - Obsessions and compulsions: normative data on the Padua Inventory from an Italian non-clinical adolescent sample. AB - In the present study the structure of obsessive-compulsive phenomena in non clinical adolescents was investigated by 'The Padua Inventory' (PI). The PI is a self-report measure of obsessive and compulsive symptoms which is used in clinical and research settings. The use of PI in adolescents has been limited by the lack of normative data. Consequently, adolescent validation has both theoretical and practical implications. PI was administered to 566 normal Italian high school subjects, ranging in age from 15 to 18 years. The mean total score of PI and the mean score of 'mental activities', 'becoming contaminated' and 'urges and worries' sub-scales points to significant differences between males and females. Females reported more obsessions and cleaning rituals than males. Males show more urges and fears than females. Moreover, our data underline that younger subjects get higher mean scores than older subjects in all scales. PMID- 10472710 TI - Stability of driving fear acquisition pathways over one year. AB - The present study was conducted in response to increasing concerns about the potential unreliability of retrospective accounts in assessing the origins of fears and phobias. Some investigators [e.g. Menzies, R.G., & Clarke, J.C. (1993). The etiology of fear of heights and its relationship to severity and individual response patterns. Behaviour Research and Therapy, 31, 355-365; Kirkby, K.C., Menzies, R.G., Daniels, B.A., & Smith, K.L. (1995). Aetiology of spider phobia: Classificatory differences between two origins instruments. Behaviour Research and Therapy, 33, 955-958; King, N.J., Gullone, E., & Ollendick, T.H. (1998). Etiology of childhood phobias: current status of Rachman's three pathways theory. Behaviour Research and Therapy, 36, 297-309.] have questioned the reliability of retrospective reports at a single assessment point, although the test-retest reliability of such accounts has yet to be examined. The aim of the present study was to conduct a one-year follow-up of the subclinical driving-fearful sample studied by Taylor and Deane [Taylor, J. E., & Deane, F. P. (1999). Acquisition and severity of driving-related fears. Behaviour Research and Therapy, 37, 435 449.] to primarily investigate, the stability of fear onset ascriptions and fear severity over time. 85 respondents completed a questionnaire which assessed fear origins, anxiety response patterns, and additional fear-relevant events occurring over the year. The results suggest that retrospective accounts of fear onset may be quite unstable over time, although this instability does not clearly appear to be related to intervening events, and limitations of the study make these results inconclusive. Fear-relevant negative thinking worsened over time, while physiological reactions and general anxiety remained relatively stable. The theoretical, methodological and clinical implications of the findings are discussed, along with suggestions for future research. PMID- 10472711 TI - The role of traumatic experiences in the genesis of obsessive-compulsive disorder. AB - This paper discusses the role of traumatic stress in the genesis of obsessive compulsive disorder. While the early classical conditioning theory of the onset of obsessive-compulsive disorder has only limited empirical support, authorities have always recognised the role of stress in precipitating this disorder, and in triggering relapse in those who have been successfully treated. Here, clinical cases are cited that show a causal link between severe trauma and the onset of obsessive-compulsive disorder. The nature of the traumatic reaction and the possible mechanisms by which it leads to frank obsessive-compulsive disorder are discussed. The apparent links between this and posttraumatic stress disorder are also explored. Finally, implications for therapy are considered. PMID- 10472712 TI - Disgust sensitivity, trait anxiety and anxiety disorders symptoms in normal children. AB - There is evidence to suggest that disgust sensitivity plays a role in the development of small animal fears and phobias. Recently, Phillips, Senior, Fahy, and David (1998) [Phillips, M. L., Senior, C., Fahy, T., & David, A. S. (1998). Disgust: the forgotten emotion of psychiatry. British Journal of Psychiatry, 172, 373-375.] suggested that disgust sensitivity is also involved in various other anxiety-based symptoms (e.g. obsessive-compulsive complaints, social phobia). The present study sought to test this suggestion in a large sample of normal school children (N = 189). Children completed a measure of disgust sensitivity, the trait anxiety version of the Spielberger State-Trait Anxiety Inventory for Children and the Screen for Child Anxiety Related Emotional Disorders, an instrument that measures DSM-defined anxiety disorders symptoms. Disgust sensitivity was indeed found to be correlated with a broad range of anxiety disorders symptoms. However, results also indicated that these correlations were predominantly carried by trait anxiety. That is, when controlling for levels of trait anxiety, only specific phobia symptoms (including animal phobia, blood injection-injury phobia and situational-environmental phobia) and separation anxiety disorder symptoms were significantly related to disgust sensitivity, although correlations were rather modest. Taken together, these findings cast doubts on the claim that disgust sensitivity is an unique and independent factor that contributes to a broad range of anxiety disorders. PMID- 10472713 TI - Philosophy of science and the progressiveness of the DSM's theory-neutral nosology: response to Follette and Houts, Part 1. AB - Follette and Houts [Follette, W. C., Houts, A. C. (1996). Models of scientific progress and the role of theory in taxonomy development: a case study of the DSM. Journal of Consulting and Clinical Psychology, 64, 1120-1132] argue on philosophy of-science grounds that the DSM's theory-neutral nosology is scientifically unprogressive because of its growing number of categories and lack of a unifying explanatory theory. They suggested replacing the DSM by competing theory-laden diagnostic manuals. I argue: (1) the ways things can go wrong with the mind are inherently diverse, so a unified theory of mental disorders is unlikely; (2) the claim that an increase in categories is inconsistent with scientific progress is empirically false; (3) the claim that the DSM's new categories expand the domain of disorder is largely false; (4) progress in a theoretically fragmented field requires a shared theory-neutrally defined domain; (5) theory-neutral diagnosis and integration of etiological theories is preferable for now to competition among theory-based diagnostic manuals; (6) philosophy of science supports use of a theory-neutral nosology for now. PMID- 10472714 TI - The concept of disorder as a foundation for the DSM's theory-neutral nosology: response to Follette and Houts, Part 2. AB - Follette and Houts [Follette, W. C., Houts, A. C. (1996). Models of scientific progress and the role of theory in taxonomy development: a case study of the DSM. Journal of Consulting and Clinical Psychology, 64, 1120-1132] argue that 'mental disorder' and 'function' are value concepts that offer no scientific basis for the DSM's theory-neutral nosology or for distinguishing disorders from other behavioral problems. They also claim that the DSM presupposes a biological theory of etiology, thus is not really theory-neutral. They recommend replacing the DSM by theory-laden diagnostic manuals. I argue: (1) DSM criteria do not imply a biological model of disorder, (2) 'mental disorder' and 'function' have scientific content that allows one to distinguish disorder from nondisorder, (3) the evolutionary 'harmful dysfunction' analysis of disorder [Wakefield, J. C. (1992a). The concept of mental disorder: on the boundary between biological facts and social values. American Psychologist, 47, 373-388] coherently demarcates disorder from nondisorder, (4) the proposed behaviorist alternative to the DSM illustrated by the articles in Follette's special section [Follette, W. C. (Ed.) (1996a). Special section on the development of theoretically coherent alternatives to the DSM system. Journal of Consulting and Clinical Psychology, 64, 1117-1201] is incoherent because it does not distinguish disorder from nondisorder. PMID- 10472715 TI - [Microbiological standardization of laboratory animals]. AB - An overview is given on various aspects of microbiological standardisation of laboratory animals. The paper focuses on small rodents because rats and mice are the most frequently used animal species. An introductory chapter describes the importance of micro-organisms and their potential effects on animals and animal experiments. Some terms describing the microbiological quality of rodents (e.g., germ-free, "SPF", conventional) are explained. Housing conditions and requirements for the management which are both necessary to maintain a high microbiological standard in the animal facility are described. The most relevant sources of micro-organisms are briefly discussed. An overview is given on some aspects which have to be considered during health monitoring like, e.g., micro organisms to be monitored, sample size and frequency, age of animals. In the reference section primarily actual and general articles or recommendations are listed. Further details can be found at internet homepages which are listed for several organisations active in laboratory animal science. PMID- 10472716 TI - [Detection of Mycobacterium avium subspecies paratuberculosis (M. p.) by the polymerase chain reaction (PCR) and culture experiments in animals in Austria]. AB - The polymerase chain reaction and cultural method was applied to detect Mycobacterium avium subspecies paratuberculosis (M. p.) in fecal samples of 165 suspected and of 35 diseased cattle, 18 intestinal tissues and 14 lymph nodes of diseased ruminants as well as organ material (n = 10) were also tested. An agreement (+/+, -/-) in the results between both methods was found in 89.7% of all cases examined. 12.7% of samples of suspected and 9.1% of diseased animals were only positive by PCR. In cases of intestinal lymph node and tissue as well as udder tissue of diseased cattle a total agreement between PCR and culture was observed. Lymph node of lung of one diseased cow was positive only in PCR. PMID- 10472717 TI - [Resistance patterns of ovine mastitis pathogens]. AB - For control of the udder health status of milk sheep the minimal inhibitory concentration of mastitis causing pathogens to 12 different chemotherapeutics was determined by means of Sensititre microplate method. 73 staphylococci- and 8 streptococci-strains was examined. Following staphylococci-strains were found: S. aureus (11), S. epidermidis (34), S. chromogenes and S. xylosus (6 each), S. hyicus and S. warneri (each 5) as well as S. sciuri and S. simulans (3 each). MIC values to penicillin, amoxycillin and ampicillin of 2 penicillinase-positive S. aureus-strains were more than 4.0 micrograms/ml. The remaining isolates and the major part of coagulase-negative staphylococci displayed MIC-values up to 1.0 microgram/ml to beta-lactamantibiotics except cloxacillin. Against cephalosporines, cephalexin excluded, and aminoglycosides very low MIC-values concerning staphylococci were observed. In case of spiramycin MIC-levels more or less than 8.0 micrograms/ml were determined. Streptococci exhibited MIC-values in the range of 0.06 to 0.5 microgram/ml against beta-lactamantibiotics with exclusion of cloxacillin, cefoperazone and cefquinome and spiramycin as well. PMID- 10472718 TI - Case report: primary fibrosarcoma of the liver in a cow. AB - An unusual case of primary fibrosarcoma of the liver in a cow is described. The most prominent pathological findings were severe abdominal haemorrhage and neoplastic masses confined to the liver only. Histologically, spindeloid cells arranged in interlacing bundles running in various directions and numerous areas of necrosis and ossification were randomly scattered in the neoplasm. PMID- 10472719 TI - Study of the effect of Lactobacillus paracasei and fructooligosaccharides on the faecal microflora in weanling piglets. AB - The influence of administration of Lactobacillus paracasei alone and mixture of Lactobacillus paracasei and fructooligosaccharide on faecal bacteria counts in the weanling pigs was investigated. The administration of Lactobacillus paracasei alone significantly decreased Clostridium (p < 0.05) and Enterobacteriaceae (p < 0.05) counts as compared to the control. Lactobacillus paracasei administered in combination with fructooligosaccharide significantly increased Lactobacillus (p < 0.01-p < 0.05), Bifidobacterium (p < 0.05), total anaerobes (p < 0.05), and total aerobes (p < 0.05) counts compared to control group as well as Lactobacillus paracasei group and significantly decreased Clostridium (p < 0.05) and Enterobacteriaceae (p < 0.01) counts compared to control group. The results obtained point out to a synergic effect of the combination of Lactobacillus paracasei and fructooligosaccharide on numbers of bacterial populations observed in the faeces of the weanling pigs. PMID- 10472720 TI - [Age dependence of the density of alpha-1 adrenergic receptors in liver cell membranes of domestic fowl]. AB - Age dependence of alpha 1-adrenergic receptor number on hepatocytes of chickens was determined by incubation of liver cell membranes from chickens of different ages with 3H-prazosin. We showed, that there is a continuous and significant rise of alpha 1-adrenergic binding sites with increasing age of the chickens from 226 +/- 22 fmol/mg protein at the age of up to 4 weeks to 628 +/- 34 fmol/mg protein in laying hens. Changes in receptor affinity could not be registered. The KD values ranged insignificantly from 0.34 +/- 0.03 nM to 0.50 +/- 0.06 nM in the different ages of chickens and amounted to 0.44 +/- 0.02 nM averagely. The classification of the binding sites for 3H-prazosin to the alpha 1B-subtype could be caused by the high sensitivity to chlorethylclonidine and by the low affinity for alpha A-adrenergic antagonists. PMID- 10472721 TI - [Wild is not really wild: brain weight of wild domestic mammals]. AB - Domestication leads to the reduction of brain weight, decreases reach from 8.1% in laboratory rats up to 33.6% in domesticated pigs. The question is: Do brain weights increase by feralization? We compared the brain weights of domesticated mammals (cat, dog, pig, goat, ass) with their feral forms. In none of the cases studied, brain weight is increased in wild domestic mammals. So, feral mammals do not return back to the status of their wild species. PMID- 10472722 TI - [Long-term EKG in dogs: comparison between a computerized system and visual arrhythmia analysis]. AB - Based on results of holter ecg recordings taken from 38 dogs the computerized analyses system turned out as method not exactly valuing the frequency and sensitivity of arrhythmias. Despite there was a significant correlation between the computerized and visual arrhythmia analysis for ventricular respectively supraventricular premature beats, in individual cases there was an important aberration between the particular values. PMID- 10472723 TI - [Changes in platelet concentrates from dogs due to storage. I. Platelet count in in vitro function]. AB - The possibility of storage of canine platelet concentrates (PC) was investigated using PC from dogs which were obtained with an automatic cell separator in C4 cell separation sets with low gasdiffusionable Polyvinylchlorid (PVC) storage containers or in C4L-sets developed for storage with high gasdiffusionable Polyolefin (PO) containers, respectively. The storage was carried out for a period of 10 days under permanent agitation at 22 degrees C (C4/22 degrees C, n = 10; C4L/22 degrees C, n = 11) or at 4 degrees C (C4L/4 degrees C, n = 6), respectively. Measurements were done directly after production of the PC, after 6 hours and then daily during the 10-day storage period. In the first part of this paper the results of platelet count (determined automatically with a blood cell differentiation automat and visually), the number of platelet aggregates, the mean platelet volume (MPV) as well as the platelet function with regard to the platelet aggregation induced by collagen or ADP and the resonance-thrombogram (RTG) are presented. The platelet count, measured automatically as well as visually, remained preponderantly constant over the complete storage time in all storage conditions. Dependent on the storage conditions--especially under storage at 22 degrees C--an increase of the number of platelet aggregates and a decrease of MPV was determined. In addition, the loss of platelet function measured by aggregation induced by collagen as well as by ADP showed a significant dependency of storage conditions. The stored platelets lost their ability to aggregate under C4/22 degrees C-conditions after a storage period of 2 days, under C4L/22 degrees C-conditions after 4 days and under C4L/4 degrees C-conditions not before 8 days of storage. Previous resuspending of platelets in fresh plasma delayed the loss of platelet function. Because the loss of platelet function described in the RTG became significant at nearly the same point in time, a storage of canine PC under corresponding conditions can be recommended for upto 2 days (C4/22 degrees C), for 4 days (C4L/22 degrees C) or 8-10 days (C4L/4 degrees C), respectively. PMID- 10472724 TI - [Breath condensate--a medium obtained by a noninvasive method for the detection of inflammation mediators of the lung]. AB - Collection of exhaled condensate (freezing of expired air under conditions of spontaneous breathing) is a non-invasive method permitting the collection of material originating from the lung and the lower respiratory tract so that it can be used for diagnostic examinations. In order to be able to evaluate the diagnostic evidence of exhaled condensate samples in cases of respiratory disease of the calf, leukotriene B4 (LTB4) in bovine exhaled condensate was determined. The influence of the breathing pattern and body temperature on the quantity of condensate to be collected was tested in a total of 49 exhaled condensate samples. It became obvious that the exhaled condensate quantity obtained per time unit is dependent on the ventilation volume per time unit (minute volume). In exhaled condensate samples from 35 clinically healthy calves, LTB4 concentrations of less than 250 pg/mL exhaled condensate were detected. A total of 14 exhaled condensate samples from 7 calves was analyzed before and after experimental respiratory infection with Pasteurella multocida D. In parallel to the analysis of LTB4 in exhaled condensate, the lung function of the calves was examined by means of impulse oscilloresistometry. The increase of LTB4 in the exhaled condensate post infection correlated significantly (p < or = 0.05) with decreases of respiratory reactance. The determination of LTB4 concentrations in exhaled condensate seems to be suitable, in principle, for the detection of inflammations in the respiratory system of the calf. Further studies are needed for the evaluation of the diagnostic validity of the method. PMID- 10472725 TI - [Percutaneous vertebral augmentation (PVA) in osteoporosis of the vertebrae--an experimental study]. AB - The present study was conducted with the aim of establishing whether minimally invasive percutaneous techniques used to stabilize osteoporotic vertebrae are technically feasible. Two different methods were investigated in human thoracolumbar cadaveric vertebrae. In the first technique, special titanium implants were placed via a postero-lateral approach. With the second method, the vertebrae were filled with different types of cement of different viscosities. After each procedure, the vertebrae were examined with conventional X-ray and CT scans. The first technique proved quite unsuccessful--the insertion of the titanium implants proving difficult despite the use of special instruments. The results achieved with the second method were much better. The use of low viscosity bone cement produced the best results. Despite a single lateral point of entry, the vertebrae were almost completely filled right into the contralateral side. Lumbar vertebrae required an average volume of cement of 7 ml (range: 6.5-10 ml) and thoracic vertebrae 5.5 ml (range: 4-7 ml). Specially developed cement application devices made possible problem-free, controlled introduction of the cement. PMID- 10472726 TI - Development of an advanced mechanised gait trainer, controlling movement of the centre of mass, for restoring gait in non-ambulant subjects. AB - The study aimed at further development of a mechanised gait trainer which would allow non-ambulant people to practice a gait-like motion repeatedly. To simulate normal gait, discrete stance and swing phases, lasting 60% and 40% of the gait cycle respectively, and the control of the movement of the centre of mass were required. A complex gear system provided the gait-like movement of two foot plates with a ratio of 60% to 40% between the stance and swing phases. A controlled propulsion system adjusted its output according to patient's efforts. Two eccenters on the central gear controlled phase-adjusted the vertical and horizontal position of the centre of mass. The patterns of sagittal lower limb joint kinematics and of muscle activation of a normal subject were similar when using the mechanised trainer and when walking on a treadmill. A non-ambulatory hemiparetic subject required little help from one therapist on the gait trainer, while two therapists supported treadmill walking. Gait movements on the trainer were highly symmetrical, impact-free, and less spastic. The weight-bearing muscles were activated in a similar fashion during both conditions. The vertical displacement of the centre of mass was bi-instead of mono-phasic during each gait cycle on the new device. In conclusion, the gait trainer allowed wheelchair-bound subjects the repetitive practice of a gait-like movement without overstraining therapists. PMID- 10472727 TI - [Mechanical effects on measuring displacement of the knee joint with the knee arthrometer KT1000 and KT2000]. AB - The KT2000 knee ligament arthrometer manufactured by the MEDmetric Corporation is used in the field of orthopedics to establish quantitatively the size of the anterior and posterior draw phenomena of the knee joint. The basis for the measurement is the Lachman test in which a force is applied to the tibia relative to the femur. The resulting displacement is limited by the cruciate ligaments. The measurements obtained for a potentially injured knee are compared with those of the healthy knee and empirical threshold values, and serve as a basis for deciding whether to operate on patients with cruciate ligament insufficiency. The subject of the present study is a design shortcoming of the KT2000 that may falsify the measured results when the device is not correctly employed. A simple mechanical modification of the KT2000 can reduce this error. In the present study, the error is quantified and we show how the modification can keep it within tolerable limits. PMID- 10472728 TI - [Mechanical study of potential ceramic implant materials for minimal invasive anterior lumbar interbody fusion]. AB - While autologous bone grafts are highly suitable for use in spinal arthrodesis, their use is also associated with problems (traumatization, complications). Ceramic bone substitute materials provide an attractive alternative for lumbar interbody spinal fusion. The aim of the present study was to investigate the mechanical properties of various types of ceramic using a specific fusion method. Ten specimens each of 7 different types of ceramic were tested using a hydraulic testing machine with two different sample holders: polyurethane foam (mechanical properties similar to cancellous bone) and aluminium. The parameters axial compression and axial torque were investigated. With the polyurethane foam holders, none of the ceramic implants failed under compression, while under axial rotation, two types of ceramic failed. With the aluminium holders, 3 ceramics showed no failure up to 25 kN under compression, while under torsion all the ceramics failed. One type of ceramic showed specific fracture properties with a higher load-bearing capacity after failure in comparison with all the other types studied. PMID- 10472729 TI - [Ceramic cups for hip endoprostheses. 6: Cup design, inclination and antetorsion angle modify range of motion and impingement]. AB - The present investigation focuses on total hip replacement using ceramic acetabular components. The relationship between the position of the cup and the range of motion (ROM) was investigated. A limited range of motion may cause impingement, which is defined as contact between the femoral neck and the rim of the acetabular cup. Impingement may result in wear, chipping, fracture or dislocation of the femoral head. Joint movements were simulated in a three dimensional CAD program. The results obtained underscore the importance of correct positioning and design of the cup for achieving a ROM as close to the physiological situation as possible. With ceramic cups, the inclination angle should not be more than 45 degrees, and the antetorsion angle between 10 and 15 degrees. If the cup is too vertical, the risk of dislocation and fracture of the ceramic increases. If, on the other hand, the angle of inclination is too small, flexion and abduction will be greatly limited. The study shows that acetabular components with non-recessed ceramic inserts should not be used. Slight recession of the insert helps to avoid impingement. The ROM is reduced and the risk of impingement appreciably increased when mushroom-shaped femoral heads (XL heads) or ceramic inserts protected by a polyethylene ring are used. PMID- 10472730 TI - [The installation of security in pathologic anatomy: the impact of prions]. PMID- 10472731 TI - Prion diseases--light at the end of the tunnel? PMID- 10472732 TI - The human prion diseases. A review with special emphasis on new variant CJD and comments on surveillance. AB - The transmissible spongiform encephalopathies or prion diseases represent a new group of diseases with unique clinical and neuropathological features, the transmission of which is both genetic and infectious. The responsible agent is unconventional and appears to be largely composed of a glycoprotein, the prion protein PrP. This is normally present on different cells. In prion diseases, it becomes converted to the pathogenic form PrPres which is resistant to proteinase and accumulates within the brain and this process is accompanied by the development of spongiform change, gliosis and neuronal loss. The human prion diseases include Kuru a progressive cerebellar degeneration with late dementia affecting Fore tribes in New-Guinea, now almost extinct, regarded as being related to cannibalism. Creutzfeldt-Jakob disease is the more frequent human prion disease. Its incidence is approximately one case per million per year. Four variants are now recognized: sporadic, familial, iatrogenic and the new variant. The latter represents a distinct clinico-pathological entity. It is now widely accepted that it is due to the same agent responsible for Bovine Spongiform Encephalopathy in cattle. Gerstmann-Straussler-Scheinker disease is a very rare inherited disorder due to a number of different mutations in the PRP gene, characterized by abundant deposits of plaque PrPres in the cerebral grey matter. Fatal familial insomnia is another inherited disorder due to a mutation at codon 178 of the PRP gene associated with methionine on codon 129 of the mutant allele. The main neuropathological change is neuronal loss in the thalamus with little or no spongiosis and usually no PrPres deposition. Following the emergence of new variant CJD in 1996, surveillance of all forms of prion diseases has been now been actively introduced in many European nations in order to determine the true incidence and geographic distribution of these rare disorders in humans. PMID- 10472733 TI - [Animal transmissible subacute spongiform encephalopathies: scrapie in sheep and bovine spongiform encephalopathy]. AB - In veterinary medecine, the field of animal Transmissible Subacute Spongiform Encephalopathies (TSSE) has been suddenly enlarged in 1986, by the dramatic outbreak of the Bovine Spongiform Encephalopathy (BSE) in the UK. Since this first observation BSE was recognised in various EU countries. BSE is largely considered, at the present time, as a food born infectious disease, resulting from the feeding of bone and meat meals (BMM) of bovine, after a rendering process modification. It is largely agreed that the ovine Scrapie prion could be transmitted to bovine through the alimentary way. Unlike Scrapie, BSE is characterized by its remarkable pathological monomorphism and the absence of obvious relationship between the prion infectivy and the recipient genotype. Another feature of BSE agent is its broad spectrum of interspecies infectivity, including through the alimentary route, raising very important questions both for animal and public health. The new variant of Creutzfeldt-Jakob disease (nv CJD) which was described in the UK, results certainly from a human contamination by bovine products. Regulation have been made in the EU, in order to stop the infectious spreading (BMM banning for Ruminants feeding) and to protect the consumer (banning of "specified risk materials" in human alimentation). PMID- 10472734 TI - [Acquired forms of Creutzfeldt-Jakob disease]. AB - AIM: To establish the recent neuropathological data concerning the acquired forms of human prions diseases at the exception of the new variant. RESULTS: Kuru transmitted by cannibalism involved mainly the cerebellum with cerebellar tracts degeneration and presence of amyloid plaques called "kuru plaques" labeled by anti PrP antibodies. To day the disease has disappeared. Iatrogenic forms have been found after corneal graft or more frequently dura-mater graft. In these later cases, neuronal loss, spongiosis and gliosis are diffuse in cerebral and cerebellar cortex with the same intensity. The cases associated with pituitary derived human growth hormone administration are observed in young patients. Histologic lesions, in 16 cases, are present in basal ganglia, thalamus, brain stem and predominantly in the cerebellum especially in the granule cell layer. Cortical involvement is moderate. Plaques have not been always looked for. CONCLUSION: The diversity of acquired forms is in favor of the hypothesis of selection of specific strains. PMID- 10472735 TI - Prion disease with octapeptide repeat insertion. AB - OBJECTIVE: About 8% of prion disease cases are familial and a few are due to an octapeptide repeat insertion (OPRI) in the prion protein gene. A suitable neuropathological examination has been performed in 20 cases from 9 families and in 3 isolated cases. The number of OPRI ranges from 4 to 9 multiples of 24 base pair. METHODS: Results from routine histopathological preparations and from immunohistochemistry performed after special tissue pretreatment were compared with those of molecular genetic investigation. RESULTS: Eight cases with 4 to 7 multiples of OPRI exhibited characteristic elongated deposits in the cerebellar molecular layer, which were visible on slides prepared with antibodies against the prion protein only. Conversely, 6 cases with 8 or 9 multiples of OPRI presented typical plaques already obvious on routine preparations. CONCLUSIONS: These variable modifications in the cerebellar molecular layer deserve to be underlined, in particular the elongated deposits which are characteristic for cases presenting 4 to 7 OPRI. PMID- 10472736 TI - Gerstmann-Straussler-Scheinker disease and the French-Alsatian A117V variant. AB - Gerstmann-Straussler-Scheinker disease is a rare familial form of prion disease. This autosomal dominant disorder is constantly associated with a point mutation on the PrP gene. Eight mutations affecting respectively codons 102, 105, 117, 145, 202, 212 and 218, have been so far described. Symptoms are variable and include ataxia and dementia. They generally appear between the fourth and sixth decade. Mean duration of the disease (5 years) is on the whole longer than that of other familial forms of prion diseases. Gerstmann-Straussler-Scheinker disease is neuropathologically characterized by the presence of numerous multicentric or unicentric PrP amyloid deposits widespread throughout the encephalon. Spongiform change is inconstant. Neurofibrillary tangles have been described in some families. Clinicopathological features show considerable variability. Pathogenesis of amyloidosis and associated lesions as well as factors underlying the phenotypic polymorphism of the disease remain only partially known. PMID- 10472737 TI - [Fatal familial insomnia]. AB - Since its description in 1986, Fatal Familial Insomnia (FFI) became the third most common inherited prion diseases (23 described families, 3 isolated cases). It is characterized by a mutation at codon 178 of the prion protein gene cosegregating with the methionine polymorphism at codon 129 of the mutated allele. Insomnia, dysautonomia, disruption of circadian rhythms and motor dysfunctions (myoclonus, ataxia, dysarthria, spasticity) are the main clinical symptoms in the homozygote patients (met/met at codon 129). Heterozygotes have motor dysfunctions from onset and cognitive changes. Pheno-typic variability does not appear to be strictly related to codon 129 polymorphism as recently stressed in some reports. Neuropathology shows marked neuronal loss and gliosis in the thalamus, especially in the medio-dorsal and antero-ventral nuclei, without any amyloid deposits. Some spongiosis may be seen essentially in the cerebral cortex, in patients with longer duration disease. The D178N mutation coupled with the 129 valine codon is linked to a subtype of Creutzfeldt-Jakob disease. However, in these two phenotypically different diseases, two protease resistant fragments of the pathogenic PrP (PrPres) are accumulated. They differ in molecular mass. In FFI PrPres, the unglycosylated form is underrepresented. This particularity does not result from the preferential conversion of the glycosylated forms but from an inaccessibility of non glycosylated form to conversion. PrPres has been shown to be form allelic origin. Neuronal apoptosis was found to contribute to neuronal loss in FFI. Its presence correlates with neuronal loss, being invariably noticed in the thalamus and medullary nuclei. It is not correlated with PrPres accumulation. The quantity of deposits is globally low in FFI brains and rarely immunohistochemically detected. Pathogenesis of lesions and clinical signs remain to be assessed. Protein dysfunction could be hypothesized according to some clinical and experimental data as well as to the discordance between protein accumulation and programmed cell death. Neurotoxicity is also postulated. Studies on this pathology led to consider the existence of "strains" in human prion diseases. Despite remarkable advances, many issues remain unsolved in this non spongiform prion disease. PMID- 10472738 TI - [A process of programmed cell death as a mechanisms of neuronal death in prion diseases]. AB - Neuronal loss is a salient feature of prion diseases; however, its cause and mechanism, particularly its relationship with the accumulation of the pathogenic, protease resistant isoform PrPres of the cellular prion protein PrPc, are still unclear. A number of studies suggest that it could occur through a process of programmed cell death which is consistent with the lack of inflammation in these conditions. In this paper, we review the different techniques used to identify apoptosis of neurons, and analyse the studies demonstrating neuronal apoptosis in prion diseases, either experimentally, in animal or in human. Apoptosis of rat hippocampal neurons, in cultures exposed to a synthetic peptide homologous to the prion protein, has been identified on morphological criteria after staining by a fluorescent marker of DNA and by gel electrophoresis of neuronal DNA. Apoptosis of neurons has also been identified in vivo using in situ end labelling and electron microscopy in scrapie infected mice. In human, apoptotic neurons were identified by in situ end labelling in Creutzfeldt-Jakob Disease and in Fatal Familial Insomnia. Apoptotic neurons were mostly found in damaged regions and their presence and abundance seemed to correlate closely with neuronal loss. Neuronal apoptosis also correlated well with microglial activation as demonstrated by the expression of major histocompatibility complex class II, antigens, and with axonal damage as identified by beta-amyloid protein precursor immunostaining. In contrast, there was no clear correlation between the topography and severity of neuronal apoptosis and the type, topography and abundance of prion protein deposits as demonstrated by immunohistochemistry. Similarly, within the framework of comparable phenotypes, there was no difference in the abundance and distribution of apoptotic neurons according to the aetiology whether sporadic, familial, or iatrogenic, of the disease. The pathogenetic mechanism of neuronal apoptosis remains speculative and several hypothesis have been proposed. The lack of a direct association between neuronal damage and PrPres deposition may support models of neuropathogenesis based on "loss of function" of PrPc, such as withdrawal of defined activation signals inducing programmed cell death, rather than neurotoxicity. It is also possible that PrPres is neurotoxic and the dissociation between neuronal damage and the amount of protein only reflects variations in selective neuronal vulnerability. Finally, neuronal apoptosis might be an indirect consequence of PrPres deposition. PrPres induced dendritic or axonal damage, perhaps enhanced by consequent microglial activation, might contribute to neuronal apoptosis either due to deafferentation or to retrograde neuronal degeneration. PMID- 10472739 TI - Protection of personnel and environment against Creutzfeldt-Jakob disease in pathology laboratories. AB - Most neuropathology laboratories have been faced with the question of dealing with cases of Creutzfeldt-Jakob disease (CJD) which is a rare neurodegenerative disorder. Neuropathologists have been long aware of the transmissibility and unique properties of the agent which make it resistant to conventional inactivating reagents. The emergence of iatrogenic cases and of the bovine spongiform encephalopathy (BSE) crisis has induced anxiety among laboratory staff and raised questions about the efficiency of safety measures and procedures hitherto applied in pathology laboratories. This article aims at presenting an overview of the risk involved in handling CJD material. It gives practical advice and a key to more detailed procedures, guidelines and recommendations available in scientific literature and through government agencies. Neuropathologists and biochemists are at a higher potential risk than others since the diagnosing of CJD involves the handling of nervous tissue which contains the highest levels of infectivity. PMID- 10472740 TI - Who provides the best retrospective information while assessing adult's premorbid functioning? A methodological study in a first-admission sample with psychosis. AB - The present study investigates the quality of data received retrospectively from subjects in their first psychotic admission and from their relatives in comparison to school records. The sample included 161 subjects out of 309 subjects (51.4%) for whom at least part of the information about the school was received. The study showed that valuable information can be extracted from school records if done systematically. Significant other's (usually the biological mother) report more accurately on academic performance during the early school years than the subject himself. The school records did not always report behavioral and social problems reported by the subject themselves or their relatives. CONCLUSIONS: 1) For retrospective information on academic performance the school records can be considered the optimal source. When records are not available, parents, in general, provide reliable information. The respondent himself can be considered highly reliable when reporting poor grades; 2) Disruptive behaviors during the early school years are reported at high frequency by school records. Parents and respondents tend to report increasing disruptive behaviors during the middle and high school years; 3) When eliciting information about social domains it is important to obtain information from all available sources. PMID- 10472741 TI - Agreement between spouses and children in descriptions of personality change in Alzheimer's disease. AB - The purpose of the present study was to evaluate the agreement among spouses and children in their describing the current and past personality of patients with Alzheimer's disease (AD) and to determine the relations between their descriptions and selected demographic and clinical variables. The subjects were 22 dementia out-patients who fulfilled the DSM-III-R criteria for uncomplicated dementia of the Alzheimer's type and the NINCDS-ADRDA criteria for AD. Spouses and children of these patients were separately administered Brooks and McKinlay's personality inventory consisting of 18 pairs of adjectives that characterize the extremes of a behavioral dimension, and were asked to judge where the subject's demeanor fell on a five-point scale, varying from +2 to -2, in which zero was regarded as neutral. Spouses and children reported marked changes in most measured behavioral dimensions following the onset of AD. Spouses identified significant changes on 14/18 items and children on 13/18 items. Spouses and children agreed on practically all items concerning personality attributes before the onset of illness, and on 16/18 items after it. Changes in personality were not correlated with the studied demographic characteristics of patients, spouses and children, nor with the cognitive deficits and illness duration of the patients. PMID- 10472743 TI - Emotional reactions to the mentally ill are positively influenced by personal acquaintance. AB - The study sought to evaluate whether personal acquaintance with a psychiatric patient increases or decreases stigmatization. MATERIAL AND METHOD: The research was carried out in a Turkish university. The Social Distance Scale and the Affective Reaction Scale were completed by final year students. The subjects were then asked whether they have ever been acquainted with a patient with a mental disorder. RESULTS: Subjects who had not been acquainted with a patient with mental disorder had a higher score on the Affective Reaction Scale than subjects who had been acquainted. CONCLUSION: This finding suggests that acquaintance with a psychiatric patient reduces negative emotions towards people with mental disorders. It is suggested that this result is related to the resolution of sensitization. PMID- 10472742 TI - Public awareness of the effectiveness of psychiatric treatment may reduce stigma. AB - Stigmatization is of importance in mental disorders. It is widely believed that one of the most important factors in stigmatization is considering the patients as dangerous. This study aims at analyzing the impact of knowledge regarding the chances of treating mental illness. MATERIALS AND METHODS: 700 final year university students filled out the Dangerousness Scale and gave their opinions on whether mental illnesses are treatable. RESULTS: The results showed that the subjects who think that the chance of treatment for mental diseases does not exist find mentally ill people more dangerous than the subjects who declare the opposite. On the basis of this finding, it can be suggested that drawing public attention to the therapeutic advances in psychiatry may lead to a decrease in stigmatization. PMID- 10472744 TI - Hand parameter differences between psychiatric patients and normal controls: a preliminary evaluation. AB - The basic assumption of this study was that it is possible to identify unusual hand features in the hands of unusual populations, and that a combination of certain unusual hand features can be associated with mental illness. One hundred and seventeen people were studied in two groups: 63 mental patients (40 males, 23 females) hospitalized in the Abarbanel Hospital, Israel, and a control group of 54 people (27 males, 27 females). Twelve hand parameters were studied, and 11 of them were found to be significantly different. Out of them, 7 show the significant result of p < 0.001. The results of this study suggest that the features of the hand may be considered as a part of the phenomenon of minor physical anomalies which are apparently neurodevelopmental markers. It suggests also the desirability of further research in this area. PMID- 10472745 TI - Advantages of the psychiatric liaison-attachment scheme in a family medicine clinic. AB - BACKGROUND: The study presents the advantages of the psychiatric liaison attachment scheme, by analyzing the psychiatric consultations of a 12 months period (1995), in an urban family medicine clinic in Israel. METHOD: Family physicians filled a questionnaire about all patients who had been seen at the consultation. RESULTS: The consulting psychiatrist saw 46 patients. The reasons for consultation were mostly to confirm a diagnosis and to decide upon medications. Medical diagnoses most often made were depression and personality disorder. The psychiatrist referred 35% of patients for further therapy in the psychiatric clinic. The family physicians would have referred 45% of patients to psychiatric clinics, if they had no psychiatric consultation available. All patients referred complied with the recommended referral. Family physicians saw accessibility as the main advantage of this consultation strategy. LIMITATIONS: This study was done in a unique setting, a teaching family medicine clinic, with nine specialized family physicians working in the psychiatric liaison-attachment scheme for as long as 10 years and more. Therefore the results of this study may not be generalized to other clinics. CONCLUSIONS: We conclude that the advantages of the psychiatric liaison-consultation method were for the patients, the family physicians and the psychiatric consultant. The main advantages for the family physicians, as stated by them, were the accessibility, the non-stigmatic availability of a psychiatrist in the clinic, and the good compliance with referral to psychiatric therapy. For the consulting psychiatrist, the advantages were the valuable information from the family physicians and the social worker, the better follow-up of patients and the team work with the family practice team. PMID- 10472746 TI - Withdrawal from clozapine: the "rebound phenomenon". AB - Clozapine is an "atypical" antipsychotic agent for treating previously resistant schizophrenic patients. Its main advantages over "typical" neuroleptics are low incidence of extrapyramidal side effects and its capacity to induce therapeutic response in previously treated refractory patients. However, withdrawal from clozapine has been observed to lead to "atypical" clinical characteristics or a "rebound phenomenon," manifested in two interwoven clinical forms: (1) psychotic exacerbation, and (2) cholinergic rebound. The underlying pathophysiological mechanism of this phenomenon is postulated to be a result of cholinergic supersensitivity. In this paper, the "rebound phenomenon" will be discussed and exemplified by three case histories in which abrupt cessation of clozapine led to serious deterioration and psychotic exacerbation, and one case in which gradual titration from the drug was employed in order to preempt this hazardous occurrence. PMID- 10472747 TI - Fenethyllin psychosis: description of two cases. PMID- 10472748 TI - Skin necrosis induced by extravasation of glycerol-containing peripheral parenteral nutrition formulation. AB - Administration of parenteral nutrition (PN) via a peripheral vein has gained support over the last decade due to serious complications associated with central venous catheterization. Extravasation and tissue necrosis have been reported with both peripheral and central dextrose-containing PN formulations. The following case report represents the first documented case of skin necrosis due to extravasation of a glycerol-containing PN formulation. Our patient's condition resolved with local therapy such as elevation of the affected extremity and cold compresses. Routine evaluation of proper catheter placement is recommended to prevent this serious morbid event, while various treatment recommendations are outlined for severe injuries that do not respond to general measures such as cold compresses. PMID- 10472749 TI - Live & then give. American Medical Association. PMID- 10472750 TI - Mississippi's medical giant: Felix Joel Underwood (1882-1959), the man who saved a million lives. PMID- 10472751 TI - Core recruitment effects in SIS models with constant total populations. AB - We consider a set of SIS models for a heterosexually transmitted disease in which there is recruitment between core and non-core subpopulations as a function of prevalence of the disease. Behavior diverges from the traditional R0 threshold behavior and yields an extra pair of endemic equilibria in one case and a limit cycle in the other. Total at-risk population is constant. PMID- 10472752 TI - Hold everything! Holding policies for protecting plasma supplies. AB - In spite of advances in testing technologies for detecting infections such as human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV), occasionally blood or plasma is collected that is potentially infectious, but is not detected as such by existing screening tests. We consider the effect of a holding policy for further reducing the number of potentially infectious units that are released for fractionation. The policy dictates a holding period during which all donated units are stored. If a donor tests positive for the infection in question at a subsequent donation, then all of that donor's units currently in storage are discarded. Otherwise, donated units are released at the end of the holding period. In the case of a single disease, we determine optimal holding periods as well as policies that are as effective as the best screening tests currently available. PMID- 10472753 TI - A theoretical model of the competition between hydrolase and carboxylesterase in protection against organophosphorus poisoning. AB - A system of coupled non-linear differential equations describing interactions between organophosphorus compounds (OPs), OP hydrolase, acetylcholinesterase (AChE), and carboxylesterase (CaE) in a single compartment was derived incorporating irreversible combination of OP with AChE, hydrolytic breakdown of OP, and irreversible combination of OP with CaE. The equations were then uncoupled, providing non-linear differential equations on AChE, CaE and OP concentrations. One steady state solution of the AChE equation provided theoretical expressions for the amounts of OP hydrolyzed, bound with CaE, and bound with AChE. Assuming that the LD50 of an OP reflects the dose that depletes AChE to a 'minimal essential' level and that a single compartment model is applicable in vivo, the steady state solution becomes an equation predicting the LD50 from rate constants, initial enzyme levels, and the allowable AChE depletion. Normalization by initial AChE concentration produced a dimensionless relationship describing an 'OP toxicity surface' that clearly demonstrates regions where hydrolysis and CaE offer protection against OP poisoning. The surface can be used to theoretically predict an LD50 given only kinetic rate constants and effective whole-body AChE and CaE levels. Predictions of LD50s of seven OPs in rats were compared with published data. The relationship was found to adequately predict published LD50s spanning 5 orders of magnitude. The OP toxicity surface relationship provides a conceptual tool for use in OP toxicity research but should be particularly useful in predicting the relative protective effects of catalytic and stoichiometric scavenger mechanisms for an OP. PMID- 10472754 TI - Global dynamics of a SEIR model with varying total population size. AB - A SEIR model for the transmission of an infectious disease that spreads in a population through direct contact of the hosts is studied. The force of infection is of proportionate mixing type. A threshold sigma is identified which determines the outcome of the disease; if sigma < or = 1, the infected fraction of the population disappears so the disease dies out, while of sigma > 1, the infected fraction persists and a unique endemic equilibrium state is shown, under a mild restriction on the parameters, to be globally asymptotically stable in the interior of the feasible region. Two other threshold parameters sigma' and sigma are also identified; they determine the dynamics of the population sizes in the cases when the disease dies out and when it is endemic, respectively. PMID- 10472755 TI - Identification of mycobacterial DNA in cutaneous lesions of sarcoidosis. AB - Sarcoidosis is a multisystemic granulomatous disease of uncertain etiology. Recently, mycobacterial DNA especially Mycobacterium tuberculosis and Mycobacterium avium complex were detected in lung tissue and bronchial lavage fluid from patients with sarcoidosis by polymerase chain reaction (PCR) assays in 30% to 50% cases. Moreover, cell wall-defective form (CWDF) acid-fast bacteria have been isolated from skin lesions of patients with sarcoidosis which were later confirmed as M. avium complex by PCR assays. CWDF acid-fast bacteria were also found to grow from the blood of 95% patients with active sarcoidosis demonstrating a mycobacterial origin similar to M. tuberculosis. In view of these reports, we investigated 20 cases of cutaneous sarcoidosis using PCR/restriction enzyme pattern analysis (PCR/REPA) to detect mycobacterial DNA from paraffin embedded skin biopsy samples. The method involves restriction enzyme analysis of nested PCR products obtained with primers encoding for the 65-KDa protein common to all mycobacteria. Using three restriction enzymes, the mycobacterial DNA from PCR product was differentiated to the species level. All the 20 cases had clinical and histologic evidence of sarcoidosis. Special stains for fungi (PAS) and mycobacteria (Fite) were negative and no foreign body was identified on polaroscopic examination in any of the cases. The cell lysates of M. tuberculosis, Mycobacterium bovis, Mycobacterium avium-intracellulare, Mycobacterium kansasii and Mycobacterium marinum from Centers for Disease Control (CDC) were used as standard control for PCR/REPA. Eight cases of foreign body granuloma, seven normal skin samples from the margin of surgical excisions and 5 cases of dermatitis were used as negative controls, and 4 cases of cutaneous tuberculosis were used as positive controls. Mycobacterial DNA was detected by PCR in 16 of the 20 cases of sarcoidosis. PCR/REPA subtyped 8 of these to M. tuberculosis complex (2 cases), M. avium-intracellulare (4 cases), M. kansasii (2 cases) while the other 8 cases were non-tuberculous mycobacteria. All four cases of cutaneous tuberculosis were positive by PCR and had a typical M. tuberculosis PCR/REPA pattern. Mycobacterial DNA was not detected in any of the negative controls. Our results demonstrated that mycobacterial DNA is present in 80% of cutaneous lesions of sarcoidosis and these mycobacteria may play a role in the pathogenesis of sarcoidosis. PMID- 10472756 TI - Hobnail hemangioma ("targetoid hemosiderotic hemangioma"): clinicopathologic and immunohistochemical analysis of 62 cases. AB - Hobnail hemangioma, also known as "targetoid hemosiderotic hemangioma", represents a distinctive, benign vascular tumor, characterized histologically by a biphasic growth pattern of dilated vascular structures in the superficial dermis lined by prominent hobnail endothelial cells, and collagen dissecting, rather narrow neoplastic vessels in deeper parts of the lesion. We analyzed the clinicopathologic and immunohistochemical features in a series of 62 cases. Patient age range was 6-72 years (median: 32 years); 34 patients were male and 25 female. Clinically, a broad variation of diagnoses ranging from hemangioma to dermal melanocytic nevus and fibrous histiocytoma was suggested. Nineteen tumors arose in the lower and 13 in the upper extremities, 12 on the back, 8 in the buttock and hip region, and one case on the chest wall. Follow-up information on 35 patients (range from 1 to 4 years; mean: 1.5 years) revealed no local recurrence nor systemic metastasis. All neoplasms were located in the dermis and showed a broad morphologic spectrum in dependence of the age of the lesions. In addition to lesions resembling cavernous lymphangioma or lymphangioma circumscriptum, neoplasms were seen with morphologic features reminiscent to retiform hemangioendothelioma, progressive lymphangioma and so-called Dabska's tumor. Immunohistochemistry performed in 28 cases showed positive staining of tumor cells for CD31 in all cases tested, whereas only 3 out of 28 cases stained completely positive for CD34. In addition 4 out of 8 cases stained positively for vascular endothelial growth factor receptor-3 (VEGFR-3). Neoplastic endothelial cells were surrounded by actin-positive pericytes in only 7 out of 27 cases tested. Hobnail hemangioma occurs more frequently in male patients and arises commonly in the extremities and the trunk. Histologic and immunohistochemcial features suggest a lymphatic line of differentiation for this distinctive vascular neoplasm. PMID- 10472757 TI - Expression and localization of thymidine phosphorylase/platelet-derived endothelial cell growth factor in skin and cutaneous tumors. AB - Thymidine phosphorylase/platelet-derived endothelial cell growth factor (TPase/PD ECGF) is a catabolic enzyme that has been shown to be chemotactic for endothelial cells in vitro and angiogenic in vivo. TPase/PD-ECGF expression is increased in a variety of tumors. In the skin, TPase is active in normal keratinocytes in vitro and in vivo. Our objective was to study the expression and localization of TPase/PD-ECGF by immunohistochemical analysis in normal skin and cutaneous tumors and to correlate this information with enzymatic activity of TPase. TPase/PD-ECGF expression was observed in keratinocytes with intense staining of the infundibulum of hair follicles but no staining of hair bulbs. Expression localized primarily to the nucleus of keratinocytes in the basal layer but was more intense and cytoplasrmic in suprabasal keratinocytes. Increased expression of TPase/PD-ECGF in differentiated cells was confirmed by in vitro studies of TPase activity. In cutaneous tumors, there was positive staining for TPase/ PD ECGF in squamous cell carcinomas (10/10), eccrine poromas (3/4), eccrine syringomas (4/4), trichoepitheliomas (1/3), and tumors of the follicular infundibulum (2/3) and melanomas (5/8). There was no staining of any intradermal nevi (0/2), basal cell carcinomas (0/10) or Merkel cell carcinoma (0/1). We conclude TPase/PD-ECGF is found throughout the epidermis and its expression increases with differentiation of keratinocytes. In cutaneous tumors, expression of TPase/PD-ECGF may be linked to the cell of origin of the tumor as well as the tumor's degree of differentiation. PMID- 10472758 TI - Immunohistochemical analysis of cytokeratin expression in apocrine cystadenoma or hidrocystoma. AB - Apocrine cystadenoma is a benign sweat gland tumor which consists of various histologic types of epithelium: epithelium with decapitation secretion, pseudostratified epithelium, and flattened epithelium. Immunophenotypes, especially the expression of cytokeratins, in 8 cases of this tumor were examined immunohistochemically, because the nature or differentiation of each epithelium is not well known. The following antibodies were used as primary antibodies: those against involucrin, smooth muscle actin, and human milk fat globulin respectively, and 15 kinds of monoclonal anti-cytokeratin antibodies. Although the staining pattern varied from case to case according to the proportion of each epithelium, these epithelia, except for the flattened epithelium, revealed invariable staining patterns. The staining pattern in the epithelium with decapitation secretion and the pseudostratified epithelium was similar to that in the secretory portion including myoepithelial cells and the junction of the excretory ductal portion with the secretory portion, respectively. However, the flattened epithelium, probably due to intraluminal pressure, showed various staining patterns similar to that in each portion of the apocrine gland. Based on the above observations, we confirmed that apocrine cystadenoma was a complex tumor differentiating into each portion of the apocrine gland, i.e. the dermal duct, the junction of the excretory ductal portion with the secretory portion, and the secretory portion. PMID- 10472759 TI - Epidermal calprotectin in drug-induced toxic epidermal necrolysis. AB - Calcium ions (Ca++) in excess alter cell viability. Their potential role in drug induced toxic epidermal necrolysis (TEN) was investigated. Thirteen TEN patients were biopsied at the site of early bullous lesions and on clinically normal looking skin at least 2 cm distant from blisters. Immunohistochemistry was applied using the mouse monoclonal antibody Mac 387 recognizing the cytosolic protein complex L1 (calprotectin). The L1 antigen is a calcium-binding protein expressed by human granulocytes, monocytes-macrophages and injured epidermis, but not by normal epidermis and other cells harboured in the skin. The majority (8/13) of TEN samples from apparently non-involved skin expressed the L1 antigen in a patch-like pattern inside the epidermis where inflammatory cells were scant or absent. As assessed by computerized image analysis of TEN bullous skin, the intensity of the L1 expression in the epidermis was not statistically correlated with the amount of the infiltrating inflammatory cells (Mac 387+ macrophages, UCLH1 + T lymphocytes and Factor XIIIa+ dendrocytes) present in the dermis and in the epidermis. Such findings suggest a key role for keratinocytes in the production of the L1 calcium-binding complex. As the L1 complex formation is a calcium-dependent process, one of the first biological events in TEN could be a dramatic increase in keratinocytes intracellular Ca++ concentration following damage by the involved drug metabolites. The ultimate toxic cell dysregulation would result from the disturbance in the intracellular Ca++ homeostasis. PMID- 10472760 TI - Neuroendocrine differentiation of a metastatic basal cell carcinoma in a patient with basal cell nevus syndrome. AB - A 48 year-old white male with basal cell nevus syndrome presented with a metastatic basal cell carcinoma with neuroendocrine features. The tumor manifested aggressive behavior, having deep local invasion and metastases to para aortic lymph nodes and bone. Neuroendocrine differentiation has rarely been associated with basal cell carcinoma. The histologic, immunohistochemical, and electron microscopic studies of this rare tumor are described. PMID- 10472761 TI - Mycosis fungoides and cutaneous Hodgkin's disease in the same patient: a case report. AB - We describe a case of a 34-year-old man who presented with a 10-year history of mycosis fungoides and recent onset of inguinal lymphadenopathy, fever, and cutaneous nodules on the right hip. Biopsy of the inguinal lymph node showed mixed cellularity Hodgkin's lymphoma. Biopsy of the pre-existing skin lesions showed mycosis fungoides, while that of the nodules on the hip revealed cutaneous Hodgkin's lymphoma. Immunohistochemical and molecular analysis demonstrated a null phenotype in the Reed-Sternberg cells of the Hodgkin's disease lesions, and a T-cell phenotype in the lymphoid cells of the mycosis fungoides lesions. He was treated with chemotherapy for Hodgkin's disease, which is in remission 11 years later. Topical nitrogen mustard and maintenance PUVA therapy have been used for the mycosis fungoides, which is also in remission. PMID- 10472762 TI - 'Tubular' structures within melanocytic proliferations: a distinctive morphologic finding not restricted to Spitz nevi. PMID- 10472763 TI - The influence of religious fasting on basal tear secretion and tear break-up time. AB - Muslims abstain from food and drink from dawn to sunset during the holy month of Ramadan. An extended strict fasting may influence tear secretion and quality. We investigated changes in basal tear secretion (BTS) and tear break-up time (BUT) at the beginning and at the end of fasting on 32 healthy male patients with a mean age of 22.3 +/- 2.9 years. Body weight and urine specific gravity were also determined to assess the extent of dehydration due to fasting. Each case lost 0.1 1.25 kg of body weight while urine specific gravity changes were statistically insignificant. Mean BTS amount was 19.9 +/- 10.9 mm at the initial examination and 19.7 +/- 11.5 mm at the end of fasting (p = 0.9). BUT values were 20.1 +/- 9.3 sec at the beginning of fasting and 23.5 +/- 14.9 sec at the end of fasting (p = 0.19). We found that religious fasting in the winter season does not seem to affect BTS and BUT values in healthy individuals. PMID- 10472764 TI - Cell adhesion molecules in subretinal fluid: soluble forms of VCAM-1 (vascular cell adhesion molecule-1) and L-selectin. AB - PURPOSE: In this study we investigated the presence of soluble VCAM-1 and soluble L-selectin-1 in subretinal fluids (SRF) of patients suffering from rhegmatogenous retinal detachment . METHOD: Subretinal fluids were collected from drainage sclerotomies during surgery from 27 patients with rhegmatogenous retinal detachment complicated by proliferative vitreoretinopathy (PVR) or uncomplicated retinal detachment. Levels of sVCAM-1 and sL-selectin-1 were quantified with ELISA. RESULTS: The mean +/- SEM values of sVCAM-1 and sL-selectin-1 were 222.2 +/- 81 ng/ml and 171.7 +/- 42.1 ng/ml, respectively. The concentrations of sVCAM 1 in patients with Grade C PVR (498.2 +/- 1703 ng/ml) were significantly different from those with Grade B PVR (45.6 +/- 16.5 ng/ml) and uncomplicated retinal detachments (19.4 +/- 12.3 ng/ml). SVCAM-1 concentration in detachments which had been present for more than 8 weeks was 738.8 +/- 431 ng/ml, significantly higher than the levels in detachments of shorter duration (132.4 +/ 47.7 ng/ml). sL-selectin-1 level in Grade C PVR (291.6 +/- 92.8 ng/ml) was higher than in uncomplicated retinal detachments (72.8 +/- 13.5 ng/ml). Significantly elevated levels of sL-selectin-1 were observed in detachments lasting more than 8 weeks (605 +/- 151.1 ng/ml) compared to those of shorter duration (96.3 +/- 13.1 ng/ml). CONCLUSION: The present study supports growing evidence that these cell adhesion molecules are involved in the inflammatory process during the development and progression of PVR. PMID- 10472765 TI - Lymphocyte subsets in conjunctival mucosa-associated-lymphoid-tissue after exposure to retinal-S-antigen. AB - PURPOSE: To evaluate the immune cell subsets in conjunctival mucosa-associated lymphoid-tissue (C-MALT) following challenge with antigen. METHODS: Ten adult female Lewis rats were studied. Five rats received one drop (5 microL) of retinal S-antigen (500 microg/mL in phosphate buffered saline, PBS) instilled into the lower fornix twice daily for 10 consecutive days. Five rats received PBS only and served as controls for the experiment. Two days after the last instillation the animals were sacrificed and the orbital contents prepared for immunohistological staining. A panel of monoclonal antibodies was used: CD5, CD4, CD8, CD25, and CD45RA. The number of positive cells were counted in sections of epibulbar, forniceal, and tarsal conjunctiva. RESULTS: There was a significant increase in the number of CD8+ T lymphocytes in the conjunctiva of animals receiving retinal S-antigen when compared to control animals. CONCLUSION: Conjunctival instillation of retinal S-antigen causes an immune response in the C-MALT with a significant increase in the CD8+ T lymphocyte subset in this tissue. This response may be involved in the induction of tolerance to the encountered antigen. PMID- 10472766 TI - Acute blindness and putaminal necrosis in methanol intoxication. AB - PURPOSE: To review the neuro-ophthalmological and radiological findings of acute methyl alcohol intoxication. METHOD: 8 acute methyl alcohol intoxication cases were evaluated. RESULTS: All patients were male and their ages varied between 21 and 55. At the initial examination, 6 to 12 days after methanol intake, visual acuity ranged from no light perception to counting fingers at 2 meters with no color perception. Bilateral dense central scotomas were detected in patients whose vision was slightly preserved. Pupillary light reactions were either absent or sluggish. In 4 cases, edema of the optic disk and the peripapillary nerve fiber layer was observed. Three months later, optic atrophy had developed. Five patients underwent magnetic resonance imaging. Bilateral putaminal hyperintense lesions on T2 weighted images were observed in 3 cases. Two patients died and autopsy permission could not be obtained. Follow-up examination 12 months later revealed optic atrophy in the other six cases, with no improvement in vision. CONCLUSION: Methanol intoxication is detrimental to health, possibly resulting in blindness and occasionally death. In association with ocular signs and the other systemic and laboratory features, the ophthalmologist should be alert to the diagnosis of methanol intoxication in which visual loss may be the only symptom. PMID- 10472767 TI - Successful management of ocular toxoplasmosis during pregnancy using combined intraocular clindamycin and dexamethasone with systemic sulfadiazine. PMID- 10472768 TI - The adjunctive use of perfluoroperhydrophenanthrene (Vitreon) in diabetic vitrectomy. AB - OBJECTIVE: To evaluate the utility and efficacy of perfluoroperhydrophenanthrene in the management of retinal detachments secondary to severe proliferative diabetic retinopathy. PATIENTS AND METHODS: Forty consecutive patients with proliferative diabetic retinopathy and retinal detachments were entered into the study at nine participating clinical centers. Perfluoroperhydrophenanthrene (Vitreon) was used as an adjunct to pars plana vitrectomy and membranectomy. RESULTS: Preoperative diagnoses included combined traction and rhegmatogenous retinal detachments in 23 eyes (57.5%), traction retinal detachments in 13 eyes (32.5%), and recurrent rhegmatogenous retinal detachments in 4 eyes (10). Vitreous hemorrhage was present in 17 eyes (42.5%). Preoperative visual acuity ranged from light perception or hand motion in 28 eyes (70%) to 5/200 or greater in 12 eyes (30%). Vitreon was primarily used to flatten the retina following relaxing retinotomy in 12 eyes (30%), to displace subretinal fluid in a posterior to-anterior direction without performing a drainage retinotomy in 15 eyes (37.5%), and to manage intraoperative complications such as iatrogenic tears in 8 (20%) and retinal dialysis in 5 eyes (12.5%). The retina flattened intraoperatively in all cases, facilitating administration of laser photocoagulation. Patients were followed for a minimum of six months (mean 13.2 months). At last follow up, the macula remained attached in 37 eyes (92.5%), including 31 (77.5%) in which the retina was totally attached. The retina remained detached in 3 eyes (7.5%). Visual acuity improved postoperatively in 20 patients (50%), was unchanged in 13 patients (32.5%), and worsened in 7 patients (17.5%). CONCLUSIONS: Perfluoroperhydrophenanthrene is a useful and effective intraoperative tool for the management of complex retinal detachments secondary to severe proliferative diabetic retinopathy. PMID- 10472769 TI - Evaluation of diabetic retinopathy by fluorophotometry. European concerted action on ocular fluorometry. AB - BACKGROUND: Fluorophotometric variables (permeability of the blood-retinal barrier (BRB) and blood-aqueous barrier (BAB), corneal autofluorescence, and lenticular light transmittance) are reported to correlate with the severity of diabetic retinopathy. This preliminary multicenter study was performed to measure these variables simultaneously in patients with type 2 diabetes mellitus and to assess which of these variables could be of help in evaluating diabetic retinopathy. METHODS: Eighty-two patients with type 2 diabetes and diabetic retinopathy were recruited in seven European university clinics. Each patient was investigated three times, at intervals of about one year. The investigations included fluorophotometric determination of corneal autofluorescence, lenticular light transmittance, and permeability of the BRB and BAB. Retinopathy was classified into four grades, using a simplified evaluation system based on the Modified Airlie House retinopathy classification and applied to color fundus slides of standard fields 1 and 2. RESULTS: Multiregression analyses revealed that only corneal autofluorescence and BRB permeability were correlated with the severity of diabetic retinopathy (P < 0.05). Corneal autofluorescence and BRB permeability as single variables were found to be indicative of severe nonproliferative retinopathy and proliferative retinopathy (sensitivity 100% and 86%, respectively, and specificity 65% and 85%, respectively). Combination of both variables increased specificity to 92%. CONCLUSIONS: This preliminary multicenter study shows that fluorophotometric variables can be measured simultaneously and reliably in patients with diabetes and that corneal autofluorescence and BRB permeability (individually or in combination) could be of help in detecting severe non-proliferative retinopathy and proliferative retinopathy. PMID- 10472770 TI - Principles of contact lens tonometry. AB - We constructed a contact lens with an integrated pressure-sensing device. It is housed in a container with three force-sensing elements, each 120 degrees apart, enabling measurement of the appositional force, i.e., the force with which the instrument is held against the eye. In part 1 of the study, the lens' precision was tested against a manometric transducer in five eye bank eyes. The second part examined pressure as a factor dependent on the appositional force, and the third part of the study investigated the correct procedure for measuring baseline eye pressure, p0. The instrument described here allows investigation of three examination parameters: (a) the measurement of p0, the pressure independent of the appositional force; (b) the continuous measurement of the intraocular pressure (IOP); (c) the measurement of the IOP dependent on the appositional force, including artificial IOP elevation. PMID- 10472771 TI - Scanning laser polarimetry in corneal topographic changes after photorefractive keratectomy. AB - PURPOSE: To examine whether, or not, corneal topographic changes after excimer laser photorefractive keratectomy (PRK) for myopia and myopic astigmatism have any influence on measurements of the retinal nerve fiber layer (NFL) with scanning laser polarimetry. METHODS: Retinal NFL thicknesses were determined by scanning laser polarimetry in 17 eyes of 13 patients with myopia and myopic astigmatism before and after PRK. Total ablation depth ranged from 26 to 71 microm. We used the relative ratios for superior and inferior NFL thicknesses which were calculated by dividing the NFL values of respective regions by the nasal value. RESULTS: The mean superior NFL ratio measured was 3.02 +/- 0.92 preoperatively, and 3.00 +/- 0.76 postoperatively. The mean inferior NFL ratio was 2.95 +/- 0.75 preoperatively, and 2.99 +/- 0.66 postoperatively. There was no statistically significant difference between preoperative and postoperative NFL measurements (Wilcoxon signed rank test, p > 0.05). CONCLUSIONS: Corneal topographic changes after PRK have no significant influence on NFL measurements by scanning laser polarimetry. Our results suggest that scanning laser polarimetry can be used as a reliable method for retinal NFL thickness measurements even after excimer laser PRK. PMID- 10472772 TI - 360 degrees retinectomy in severe anterior PVR and PDR. AB - We reviewed 30 cases of 360 degrees retinectomy in advanced anterior PVR or PDR with anterior traction. The preoperative visual acuity was light perception or hand motions. In all cases, a 360 degrees retinectomy with silicone oil tamponade was performed. After a mean follow-up of 10 months, the retina was attached in 83% and the visual acuity had improved in 47% of the patients. The mean visual field in patients with a visual acuity of more than counting fingers extended to 29 degrees nasally and 38 degrees temporally. Hypotony was seen in 6 patients, of whom two developed keratopathy. In conclusion, eyes with severe PDR or PVR and a preoperative low visual acuity can be treated successfully with 360 degrees retinectomy when conventional methods have failed. The patients can regain an ambulatory visual acuity and a visual field. PMID- 10472774 TI - Relapse after complete response to anthracycline-based combination chemotherapy in metastatic breast cancer. AB - In this study we examined the clinical characteristics of relapse before and after complete clinical response (CR) to anthracycline-based combination chemotherapy in metastatic breast cancer (MBC). Our goal was to determine whether similar clinical trends could be observed during first relapse and relapse after CR. Two hundred and sixty-three patients with MBC were identified who had achieved CR after anthracycline-based combination chemotherapy for first recurrence. From this group, 226 patients had relapse after CR after a minimum follow-up of 13 years. Clinical features of their disease at first relapse and after progression from CR (second relapse) were examined, including disease-free interval (DFI) from diagnosis until first relapse, sites of recurrence, response to subsequent therapy, and survival after progression from CR. There was a significant correlation between duration of CR and survival after progression from CR. Patients who relapsed < 12 months after achieving CR had a median survival of 8.8 months after second relapse, whereas those who relapsed beyond 3 years had a median survival of 21.5 months (p = 0.0034). Neither the duration of CR nor length of survival after second relapse was related to the length of initial DFI. Patients with a short duration of CR (< 12 months) more often experienced second recurrences at multiple sites and in visceral organs (62% and 75%, respectively) than did patients with prolonged CR (> 36 months) (27% and 45%, respectively, p < 0.001). The anatomic location of metastatic disease at the time of first and second relapses was similar within a CR duration group but different among the groups. Short CR duration was associated with more frequent recurrence at visceral sites and also with chemotherapy-resistant second relapse. In conclusion, prolonged CR is associated with long survival after second relapse; however, neither CR duration nor survival after second relapse is related to the length of initial DFI. This suggests that chemotherapy, when it induces CR, may change the pace of disease progression. The tissue pattern of recurrence appears to be similar between first and second relapse, suggesting that the cellular predilection for metastatic sites has been preserved. PMID- 10472773 TI - Combined central retinal artery and vein occlusion in a child with systemic non Hodgkin's lymphoma. AB - PURPOSE: To report on a case of systemic non-Hodgkin's lymphoma and unilateral combined central retinal artery and vein occlusion. METHOD: We examined a 14-year old boy who experienced a sudden unilateral visual loss five months after the initial diagnosis of systemic non-Hodgkin's lymphoma. RESULT: Visual loss was due to combined central retinal artery and vein occlusion in association with tumoral optic nerve involvement. CONCLUSION: Although very rare systemic non-Hodgkin's lymphoma may present with central retinal artery and vein occlusion prior to overt central nervous system involvement. PMID- 10472775 TI - Cytokine-regulated urokinase-type-plasminogen-activator (uPA) production by human breast fibroblasts in vitro. AB - It has been shown that, in breast stroma, urokinase-type plasminogen activator (uPA) mRNA is predominantly expressed by myofibroblasts located at the invasive areas of the tumor. To examine which factors present in a tumor environment are candidates responsible for the induction of these uPA-producing myofibroblasts, we studied in vitro the capacity of a paired panel of normal and tumor-derived human breast fibroblasts to produce uPA protein and the myofibroblast marker alpha-smooth-muscle-actin (alpha-SMA) in response to various cytokines implicated in the process of tissue-remodeling during malignant transformation. We found that fibroblasts produced increased amounts of uPA protein after exposure to a FGF, b-FGF, EGF, PDGF-BB, and IFN-gamma, were unaffected in this respect by IL-6, M-CSF, GM-CSF and Oncostatin M, and produced decreased amounts of uPA protein after exposure to IL-1alpha, TNF-alpha, IGF-I, and IGF-II. None of these cytokines were able to induce a striking increase in the fraction of alpha-SMA positive fibroblasts. On the other hand, 25 pM TGFbeta1 increased the fraction of alpha-SMA-positive fibroblasts 5-fold in both normal and tumor-tissue-derived fibroblasts. Nonetheless, the normal-derived fibroblasts were unaffected in their uPA-producing capacity by TGFbeta1, and the tumor-derived fibroblasts produced decreased amounts of uPA protein after exposure to this cytokine, implying that at least in vitro the myofibroblast phenotype is not a prerequisite for the production of uPA by human breast fibroblasts. In addition, we established that the basal-uPA-production of both normal and tumor-derived fibroblasts was increased by autocrinely produced b-FGF-like activity, and that the basal-uPA production of at least the normal-derived fibroblasts was decreased by autocrinely produced IGF-like activity. Altogether, our data suggest an active role for fibroblasts in the process of uPA-directed breast tumor proteolysis. PMID- 10472776 TI - Genetic anticipation and breast cancer: a prospective follow-up study. AB - Genetic anticipation is characterized by an earlier age of disease onset, increased severity, and a greater proportion of affected individuals in succeeding generations. The discovery of trinucleotide repeat expansion (TRE) mutations as the molecular correlate of anticipation in a number of rare Mendelian neurodegenerative disorders has led to a resurgence of interest in this phenomenon. Because of the difficulties presented to traditional genetics by complex diseases, the testing for genetic anticipation coupled with TRE detection has been proposed as a strategy for expediting the identification of susceptibility genes for complex disorders. In the case of breast cancer, a number of previous studies found evidence consistent with genetic anticipation. It is known that a proportion of such families are linked to either BRCA1 or BRCA2, but no TRE mutations have been identified. It has been shown that the typical ascertainment employed in studies purporting to demonstrate genetic anticipation combined with unadjusted statistical analysis can dramatically elevate the type I error. We re-examine the evidence for anticipation in breast cancer by applying a new statistical approach that appears to have validity in the analysis of anticipation to data ascertained from a recent follow-up of a large prospective cohort family study of breast cancer. Using this approach, we find no statistically significant evidence for genetic anticipation in familial breast cancer. We discuss the limitations of our analysis, including the problem of adequate sample size for this new statistical test. PMID- 10472777 TI - Differential regulation of normal and tumoral breast epithelial cell growth by fibroblasts and 1,25-dihydroxyvitamin D3. AB - Mesenchymal-epithelial interactions are of paramount importance during normal and tumoral breast developments. We have investigated the paracrine growth regulation of normal and tumoral breast epithelial cells by fibroblasts derived from normal or pathological breast tissues. In some cases, breast cancer MCF-7 cells or normal epithelial cells in primary culture were cocultured with fibroblasts in a Transwell system allowing diffusible factor exchanges. Alternatively, conditioned medium produced by fibroblast cultures was added to epithelial cell cultures. Fibroblasts were shown to stimulate the proliferation of normal and carcinoma cells through paracrine mechanisms. However, the paracrine exchanges appeared to be different in normal versus tumoral breast epithelial cell growth regulation. Moreover, vitamin D-related compounds that have been proposed as anti-tumoral drugs were studied for their ability to affect normal and tumoral mammary epithelial cell proliferation and to interfere with the growth-regulatory activity of fibroblasts. Whereas vitamin D compounds inhibited MCF-7 cell growth, they led to a marked stimulation of the proliferation of normal mammary epithelial cells. Moreover, it was shown that the vitamin D analog EB 1089 can block the mitogenic effect of fibroblast-conditioned medium on tumoral but not normal breast epithelial cells. The differential effects of vitamin D compounds on cell proliferation provide further data in favor of the different behaviours of normal and tumoral mammary epithelial cells. The potential therapeutic use of vitamin D derivatives in the treatment of breast cancer is supported by these results but their growth-stimulatory properties on normal epithelial cells cannot be overlooked. PMID- 10472778 TI - Alpha2-adrenergic effect on human breast cancer MCF-7 cells. AB - (-)Epinephrine (Epi) and (-)Norepinephrine (NEpi) significantly stimulated tritiated Thymidine incorporation in MCF-7 cells at concentrations 10-30pM to 10nM, with an EC50 of 10pM for Epi and 14.2pM for NEpi. To characterize this action, cells were incubated in the presence of NEpi or Epi and different antagonists. The beta-adrenergic antagonist Propanolol showed no effect on the agonist's stimulation, whereas the alpha-adrenergic antagonist Phentolamine, reverted it completely at high concentrations (100 microM). The alpha1-adrenergic antagonist Prazosin (Pra) acted only at high concentrations, while the alpha2 adrenergic antagonist Yohimbine (Yo) reverted the stimulation at an EC50 of 0.11 microM. Likewise, when the cells were incubated in the presence of the specific alpha2-adrenergic agonist Clonidine (Clo), Thymidine incorporation was significantly stimulated at an EC50 of 0.298 pM. Again, the incubation of the cells in the presence of the alpha1-adrenergic antagonist Pra exerted its action at high concentrations, whereas the alpha2-adrenergic antagonist Yo showed a clear reversal of the agonist's enhancement at an EC50 of 0.136 microM. Moreover, Clo caused a clear and significant inhibition of stimulated cAMP levels both in the intracellular and the extracellular fractions. Yo showed a complete reversion of cAMP levels to control values in the presence of Clo, while Pra had the opposite effect. These data suggest that the stimulation provoked in Thymidine incorporation by the agonists Epi, NEpi, and Clo is, at least in part, due to an alpha2-adrenergic mechanism directly on tumoral cells, and that the effect is coupled with inhibition of cAMP levels, as described for this kind of receptors. PMID- 10472779 TI - Maintenance treatment with medroxyprogesterone acetate in patients with advanced breast cancer responding to chemotherapy: results of a randomized trial. Essen Breast Cancer Study Group. AB - The purpose of this randomized phase III trial was to study whether medroxyprogesterone acetate (MPA) maintenance treatment prolongs the time to progression in advanced breast cancer patients responding to an induction chemotherapy. Patients with progressive advanced breast cancer previously untreated with anthracylines and progestins were given epirubicin (30 mg/m2) and ifosfamide (2 g/m2) on days 1 and 8 at 3-weekly intervals. Patients without disease progression after 6 cycles of chemotherapy were randomly assigned to receive, until progression, either no treatment or MPA at a daily total dose of 500 mg. Ninety patients were randomized: 46 to the MPA arm and 44 to the observation arm. Median time to progression was longer in the MPA arm: 4.9 months versus 3.7 months in the intent-to-treat analysis (p = 0.02), and 4.9 months versus 3.0 months in the secondary efficacy analysis (p = 0.012). Seven patients were removed from MPA due to side effects. The changes in patient-rated quality of life scores were similar in both groups. The median length of survival from randomization was 17.4 months for patients receiving MPA and 18.3 months for patients randomized to observation (p = 0.39). In conclusion, in patients with advanced breast cancer achieving remission or non-progression with 6 cycles of epirubicin and ifosfamide chemotherapy, MPA maintenance treatment led to a significant, though modest, prolongation of the time to progression without affecting overall survival of the study patients. PMID- 10472780 TI - Prognostic significance of Ki-67 and topoisomerase IIalpha expression in infiltrating ductal carcinoma of the breast. A multivariate analysis of 863 cases. AB - To evaluate the prognostic relevance of Ki-67 and topoisomerase IIalpha expression in relation to tumor stage, grade, and hormone receptor content, 942 ductal infiltrating carcinomas of the breast were examined by means of the monoclonal antibodies Ki-S11 (Ki-67) and Ki-S4 (topoisomerase IIalpha). pS2, c erbB2, and p53 were additionally considered as prognostic variables. The median follow-up time was 149 months. Eight-hundred-and-sixty-three tumors reacted with Ki-S11 and Ki-S4; the labeling indices of the two antigens were closely associated (r = 0.93). Both correlated positively with the tumor size, c-erbB2, and p53 expression, and negatively with patient age, hormone receptor content, and pS2 immunostaining. In the univariate analysis, Ki-S11 and Ki-S4 scores, nodal status, tumor size, tumor grade, and progesterone receptor content strongly predicted both overall and metastasis-free survival (p < 0.00001). Estrogen receptor status, p53, and c-erbB2 were of minor significance. Concerning overall survival, multivariate Cox regression analysis selected a Ki-S4 score >25% (p < 0.00001) next to the nodal status, and before tumor size, progesterone receptor content, and patient age. Independent predictors of the occurrence of distant metastases were nodal status, Ki-S4, tumor size, grade 1, and progesterone receptor negativity, in that order. The Ki-S11 score was of independent prognostic significance only if examined as a continuous variable. We conclude that topoisomerase IIalpha expression as assessed by monoclonal antibody Ki-S4 may add valuable information to current prognostic models for breast cancer. Its predictive value appears to be essentially related to the proliferative activity of tumor cells. PMID- 10472781 TI - Differential effects of chemotherapeutic agents on the Bcl-2/Bax apoptosis pathway in human breast cancer cell line MCF-7. AB - The present study explored the effects of three commonly used chemotherapeutic agents on the Bcl-2/Bax apoptosis pathway and the interaction of these chemotherapeutic drugs with the estradiol-mediated regulation of this pathway. Our results showed that: (1) Treatment of MCF-7 cells with Adriamycin resulted in time- and concentration-dependent decreases in Bcl-2 and increases in Bax mRNA and protein levels. (2) Camptothecin elicited similar trends on Bcl-2 and Bax as Adriamycin, while etoposide, at 50-100 fold (1-5 microM) the effective concentration of Adriamycin and camptothecin, only resulted in an increase in Bax mRNA levels. (3) Adriamycin and camptothecin, but not etoposide, were effective in suppressing estradiol-stimulated increases in Bcl-2 mRNA levels. Our study provides evidence that the Bcl-2/Bax apoptosis pathway may be differentially regulated by chemotherapeutic agents. In addition, interaction between these agents and estradiol on the Bcl-2/Bax apoptosis pathway may also exist. PMID- 10472782 TI - Mutation analysis of the putative tumor suppression gene PTEN/MMAC1 in sporadic breast cancer. AB - PTEN/MMAC1, a potential human tumor suppressor gene, has been found to have inactivating mutations in several types of cancer, including breast cancer. The incidence of breast cancer in Chinese is quite low in comparison with Caucasians, and genetic factors may play some roles. To further determine the role of PTEN/MMAC1 in breast cancer in Chinese, we used loss of heterozygosity (LOH), single strand conformation polymorphism (SSCP) with direct sequencing of variant bands, and Southern blot analysis methods to analyze mutations in PTEN/MMAC1 in 52 cases of breast cancer. None had LOH at chromosome 10q23.3. One mutation was identified, a somatic 3-base deletion, in one case. Our results suggest PTEN/MMAC1 does not play a major role in the development of sporadic breast cancer. PMID- 10472783 TI - Phase II of doxorubicin/taxol in metastatic breast cancer. Argentine Multicenter Taxol Group. AB - PURPOSE: To assess the response rate, survival, and toxicity of Taxol (paclitaxel) as 1-h infusion plus doxorubicin as first-line treatment for patients with metastatic breast cancer (MBC). PATIENTS AND METHODS: Seventy-six patients with untreated MBC were recruited. All of them had measurable disease and were evaluable for toxicity. Fifty-five percent of the patients had visceral involvement. The dose of doxorubicin was fixed at 50 mg/m2 as a short intravenous infusion, followed by 200 mg/m2 of Taxol as a 1-h intravenous infusion. Doxorubicin was administered during the first seven cycles, continuing with Taxol only up to a maximum of ten cycles. RESULTS: Neutropenia was the most important toxicity: 30% grade 3 and 18% grade 4. Only 2 patients showed a decrease in the left ventricular ejection fraction (LVEF) which caused discontinuing the treatment. No clinical congestive heart failure (CHF) was observed. Seventy-four patients were eligible for response evaluation: 10 (14%) achieved complete response (CR) and 46 (62%) achieved partial response (PR). The mean duration of response was 13.47+/-1.35 months (95% confidence interval (CI): 10.82; 16.12) and the mean survival was 21.50+/-1.42 months (95% CI: 18.72; 24.29). CONCLUSION: The overall response (OR) rate was 76%. No CHF was assessed and 2 patients stopped treatment due to LVEF decrease. Although doxorubicin 50 mg/m2 followed by Taxol 200 mg/m2 in 1-h intravenous infusion presents a toxicity profile which demands a close follow-up, it represents a convenient outpatient schedule with similar activity rate compared to longer Taxol infusions. PMID- 10472784 TI - Contrast-enhanced computed tomography detection of occult breast cancers presenting as axillary masses. AB - Some non-palpable breast cancers presenting as axillary metastases (occult breast cancer, OBC) are not clinically detectable by either mammography (MMG) or ultrasonography (US). We performed contrast-enhanced computed tomography (CE-CT) in order to locate the primary tumors in five cases of OBC and succeeded in locating all of them. PMID- 10472785 TI - Reduction of polyphenol and phytic acid content of pearl millet grains by malting and blanching. AB - This work was undertaken to evaluate the changes in polyphenol and phytic acid content in malted and blanched pearl millet grains. For malting, grains were steeped for 16 hours, germinated for 48 or 72 hours and then kilned at 50 degrees C for 24 hours. Blanching was done for 30 seconds in boiling water at 98 degrees C. Results indicated that blanching resulted in significant reduction in polyphenol (28%) and phytic acids (38%). Destruction of polyphenols (38 to 48%) and phytic acid (46 to 50%) was significantly higher in grains subjected to malting than blanching: The overall results suggested that malting with 72 hours of germination was most effective in reducing the antinutrient levels of pearl millet grains. PMID- 10472786 TI - Variability in the antinutritional constituents in greengram Vigna radiata. AB - Five greengram varieties, viz. Mg 161, M3, Co2, Pusa 8793 and Pusa baisakhi, were analyzed for phytic acid, tannic acid and trypsin activity. Significant varietal variation in tannin (310 to 400 mg percent), phytic acid (201.33 to 265.33 mg percent) and trypsin inhibitor activity (55.74 to 97.70 TIU/mg) were observed. PMID- 10472787 TI - Reduction of some toxicants in Icacina mannii by fermentation with Saccharomyces cerevisiae. AB - The effect of fermentation on the level of some toxic components (hydrocyanic acid, phytic acid and oxalic acid) of Icacina mannii paste was investigated. The results indicate that fermentation caused a decrease in the level of these toxicants ranging from 178 mg/kg to 70 mg/kg for hydrocyanic acid, 638 mg/kg to 463 mg/kg for oxalic acid and 49 mg/kg to 21 mg/kg for phytic acid. When the paste was supplemented with active cells of Saccharomyces cerevisiae there was a marked decrease in the level of the three toxicants ranging from 84 mg/kg to 16 mg/kg for hydrocyanic acid, 374 mg/kg to 88 mg/kg for oxalic acid and 24 mg/kg to 5 mg/kg for phytic acid. Supplementing the paste with yeast cells at a concentration of 7.5 g/kg before fermenting, resulted in approximately a 90% reduction in the level of the three toxic compounds after 144 hours of fermentation. PMID- 10472788 TI - The effects of sun-drying surfaces and packaging materials on the storability of cowpea (Vigna unguiculata) seed. AB - The effects of sun-drying cowpea seeds for three hours on cement (CS), wood (WS), and corrugated iron sheet (CIS) surfaces and packaging separately in polyethene and jute bags were studied. Moisture-gain, and resistance to insect and mold damage were monitored monthly for six months, while proximate analyses on day zero and at the 6th month were conducted. Results showed that the three-hour sun drying of cowpea seeds on reflective surfaces (CIS and CS) enhanced the rate of moisture-reabsorption during storage and consequently, the degree of mold and insect damage irrespective of the packaging material employed. Crude fat, fiber and ash contents, unlike protein and moisture contents, remained virtually constant (p < or = 0.05). In this study, the wood surface and polyethene bag were the most preferred treatments to store sun-dried cowpea seeds for about 5.7 months. PMID- 10472789 TI - Proximate composition and some functional properties of extrusion cooked soybean and sweet potato blends. AB - Mixtures of sweet potato flour and soy flour were made in a pilot mixer. They were moisturized with 18, 25, and 30% water and extruded in a single screw extruder at 80 rpm, using a die of 6mm. Extrusion temperature was maintained at 100 +/- 3 degrees C. Effects of adding soy flour into sweet potato flour, as well as variation in feed moisture on the composition and some functional properties of the extrudates were investigated. Increase in sweet potato content increased carbohydrate values. Protein increased with increase in soy flour. Feed moisture did not significantly (p < or = 0.05) affect extrudate composition. Increase in sweet potato content and feed moisture increased expansion ratio. Bulk density decreased with decrease in feed moisture, but increased with increase in soy flour. Starch content increased as sweet potato content increased. Degree of gelatinization increased with sweet potato content. Lower feed moisture enhanced gelatinization. Water absorption index (WAI) increased as sweet potato content increased. Feed moisture had a slight effect on WAI and water solubility index (WSI). Amylose increased with increase in sweet potato content. Increase in soy flour led to an increase in yellowness (b*) of extrudates. PMID- 10472790 TI - Coconut kernel protein modifies the effect of coconut oil on serum lipids. AB - Feeding coconut kernel along with coconut oil in human volunteers has been found to reduce serum total and LDL cholesterol when compared to feeding coconut oil alone. This effect of the kernel was also observed in rats. Since many plant proteins have been reported to exert a cholesterol lowering effect, a study was carried out on the effect of isolated kernel protein in rats. Feeding kernel protein resulted in lower levels of cholesterol, phospholipids and triglycerides in the serum and most tissues when compared to casein fed animals. Rats fed kernel protein had (1) increased hepatic degradation of cholesterol to bile acids, (2) increased hepatic cholesterol biosynthesis, and (3) decreased esterification of free cholesterol. In the intestine, however, cholesterogenesis was decreased. The kernel protein also caused decreased lipogenesis in the liver and intestine. This beneficial effect of the kernel protein is attributed to its very low lysine/arginine ratio 2.13% lysine and 24.5% arginine. PMID- 10472791 TI - Effect of infection by Rhizopus oryzae on biochemical composition of stored potato tubers. AB - The carbohydrate fractions, protein and lipid contents of two cultivars of potato namely, Irish Cobbler and Red Pontiac, were altered quantitatively by Rhizopus oryzae during the 10-day incubation period. Glucose content increased during the incubation period for both healthy and inoculated tubers. Starch, maltose, sucrose, protein and lipid contents decreased more rapidly in inoculated tubers than in healthy tubers in both cultivars. The depletion in starch and protein in the infected tubers appeared to be greater for Irish Cobbler than in Red Pontiac; decreases for other constituents seemed fairly comparable. PMID- 10472792 TI - Proximate composition and functional properties of four cultivars of bambara groundnut (Voandezeia subterranea). AB - Proximate composition and selected functional properties of four cultivars of bambara groundnut (Voandezeia subterranea Thoura) were ascertained. Crude protein ranged from 17.5 to 21.1 percent; crude fat 7.3-8.5 percent; total ash 4-5 percent; crude fiber 1.8-2.0 percent; carbohydrate and moisture content for the different cultivars were 53.0-60.8 percent and 7.5-12.3 percent, respectively. The results of functional property determinations indicated that the bulk density ranged from 0.65 to 0.75 g/ml; water binding capacity 2.1-2.9 g/2g sample; oil binding capacity 0.9-1.6 g/2g sample; emulsifying activity 55.1-60.0 percent and emulsifying stability 10-12 percent. The results show that bambara groundnut has great potential for incorporation into various human foods where it could provide useful plant proteins. PMID- 10472793 TI - Effect of varied fermentation periods on the diabetogenic potential of toasted cassava granules. AB - The effects of exclusive gari diets prepared using varied fermentation periods on the fasting blood glucose levels (FBGL) and glucose tolerance (GT) of rats were studied. Thirty growing male albino rats divided randomly into 5 groups of 6 rats each were used. The 5 groups were each fed gari diets fermented for 0, 24, 48, 72 hours; and a standard rat diet, respectively, for 8 weeks. Data on the total cyanogen content and % crude protein of the diets; FBGL, GT, body weight, water:feed consumption ratio and clinical observations of the rats were collected. Results showed that the total cyanogen content and % crude protein of the diets were depleted as fermentation periods increased. The FBGL of all the gari-fed rats were significantly elevated and their GT significantly impaired with a significant variation between the groups. The elevated FBGL and impaired GT which were found to increase with increase in fermentation period were strongly inversely correlated to the % crude protein content of the diets (r = 0.92 and -0.96 respectively) suggesting that the diabetogenic potential of gari diets strongly depended on its % crude protein content. Shortened fermentation periods leading to production of gari with high total cyanogen content did not induce higher diabetogenicity. PMID- 10472794 TI - Effect of dietary rosemary extract on cell-mediated immunity of young rats. AB - The impact of rosemary extract on splenic mononuclear cell proliferation was determined. Weanling male Sprague-Dawley rats were fed diets containing 0, 100, 200 or 400 ppm rosemary extract or 400 ppm butylated hydroxytoluene (BHT) in combination with 10 or 20% casein enriched diets for 8 weeks. Splenic mononuclear cells were isolated from these animals and mitogenic response to Concanavalin A (Con A), Phytohemagglutinin (PHA) and lipopolysaccharide was determined. Con A and PHA-stimulated proliferation of spleen cells from rats fed 10% casein and 200 ppm rosemary extract was significantly higher than that of cells from the corresponding control animals. However, other levels of rosemary at 10% dietary casein or rosemary at any concentration fed along with 20% dietary casein had no impact on the mitogenic stimulation of splenic mononuclear cells. Thus, these results suggest that the use of rosemary might not have a generalized immunoenhancing effect, and will probably be effective in some stressed conditions, such as protein or antioxidant deficiency. PMID- 10472795 TI - The relationship between soil copper content and copper resistance in yeast of an ultisol in midwestern Nigeria. AB - Yeast isolates were obtained from the brewer's stock of the Guinness brewery in Benin City, midwestern Nigeria, and from local sources such as mango and sugar cane. The fruits were exposed to the immediate soil environment in eleven selected sites in the Ugbowo and Ikpoba Hill areas of Benin City, Edo State of midwestern Nigeria. A total of twenty-seven yeast isolates were obtained. Fourteen isolates were identified as Saccharomyces cerevisiae and 13 as S. carlsbergensis. Analysis of resistance to copper sulphate revealed that the two brewer's yeast isolates were resistant to 3.5 mM and 4.2 mM CuSO4, respectively, while the local yeasts were resistant to higher concentrations ranging from 6.5 to 16.5 mM. The percentage copper sulphate resistance of yeast isolates obtained from Mangifera inidica ranged from 52.4 to 73.8%, while the percentage copper sulphate resistance of yeast isolates obtained from Saccharum officinarum ranged from 51.8 to 62.6%. The two brewer's yeast isolates (BY2 and BY4) had a percentage CuSO4 resistance of 32.4 and 38.5%, respectively. Comparison of soil copper concentration and copper resistance levels in selected yeast isolates indicated that the soil copper concentrations generally influenced the ability of yeasts to resist copper. PMID- 10472796 TI - The processing alpha1,2-mannosidase of Saccharomyces cerevisiae depends on Rer1p for its localization in the endoplasmic reticulum. AB - The yeast alpha1,2-mannosidase Mns1p is involved in N-linked oligosaccharide processing in Saccharomyces cerevisiae by converting Man9GlcNAc2 to a single isomer of Man8GlcNAc2. alpha1,2-Mannosidase is a 63 kDa type II resident membrane protein of the endoplasmic reticulum that has none of the known endoplasmic reticulum localization signals (HDEL/KDEL, KKXX, or RRXX). Using antibodies against recombinant alpha1,2-mannosidase, indirect immunofluorescence showed that alpha1,2-mannosidase localization is abnormal in rer1 cells and that the alpha1,2 mannosidase localizes in the vacuoles of rer1/deltapep4 cells whereas in wild type and deltapep4 cells it is found in the endoplasmic reticulum. 35S-labeled cell extracts were subjected to double immunoprecipitation, first with antibodies to alpha1,2-mannosidase, then with either alpha1,2-mannosidase antibodies or antibodies to alpha1,6-mannose residues added in the Golgi. The labeled proteins were examined by autoradiography after sodium dodecyl sulfate polyacrylamide gel electrophoresis. A significant proportion of the labeled alpha1,2-mannosidase was immunoprecipitated by alpha1,6-mannose antibodies in wild-type, deltapep4 and rer1/deltapep4 cells with endogenous levels of alpha1,2-mannosidase, and in wild type, deltapep4, rer1 and rer1/deltapep4 cells overexpressing alpha1,2 mannosidase. The alpha1,2-mannosidase of rer1/deltapep4 cells had a slower mobility on the gels than alpha1,2-mannosidase precipitated from wild-type or deltapep4 cells, indicating increased glycosylation due to transport through the Golgi to the vacuoles. It is concluded that the endoplasmic reticulum localization of alpha1,2-mannosidase in wild-type cells depends on Rer1p for retrieval from an early Golgi compartment. PMID- 10472797 TI - Alpha-mannosidases involved in N-glycan processing show cell specificity and distinct subcompartmentalization within the Golgi apparatus of cells in the testis and epididymis. AB - The Golgi apparatus is enriched in specific enzymes involved in the maturation of carbohydrates of glycoproteins. Among them, alpha-mannosidases IA, IB and II are type II transmembrane Golgi-resident enzymes that remove mannose residues at different stages of N-glycan maturation. alpha-Mannosidases IA and IB trim Man9GlcNAc2 to Man5GlcNAc2, while alpha-mannosidase II acts after GlcNAc transferase I to remove two mannose residues from GlcNAcMan5GlcNAc2 to form GlcNAcMan3GlcNAc2 prior to extension into complex N-glycans by Golgi glycosyltransferases. The objective of this study is to examine the expression as well as the subcellular localization of these Golgi enzymes in the various cells of the male rat reproductive system. Our results show distinct cell-and region specific expression of the three mannosidases examined. In the testis, only alpha mannosidase IA and II were detectable in the Golgi apparatus of Sertoli and Leydig cells, and while alpha-mannosidase IB was present in the Golgi apparatus of all germ cells, only the Golgi apparatus of steps 1-7 spermatids was reactive for alpha-mannosidase IA. In the epididymis, principal cells were unreactive for alpha-mannosidase II, but they expressed alpha-mannosidase IB in the initial segment and caput regions, and alpha-mannosidase IA in the corpus and cauda regions. Clear cells expressed alpha-mannosidase II in all epididymal regions, and alpha-mannosidase IB only in the caput and corpus regions. Ultrastructurally, alpha-mannosidase IB was localized mainly over cis saccules, alpha-mannosidase IA was distributed mainly over trans saccules, and alpha-mannosidase II was localized mainly over medial saccules of the Golgi stack. Thus, the cell-specific expression and distinct Golgi subcompartmental localization suggest that these three alpha-mannosidases play different roles during N-glycan maturation. PMID- 10472799 TI - Intracellular trafficking of dense granule proteins in Toxoplasma gondii and experimental evidences for a regulated exocytosis. AB - The dense granules of the intracellular protozoan Toxoplasma gondii are secretory vesicles that play a major role in the structural modifications of the parasitophorous vacuole (PV) in which the parasite develops. The biogenesis of dense granules as well as the regulatory mechanisms controlling their specific exocytosis are still poorly understood. In this paper, we analyzed the secretory pathway of dense granule proteins (GRA proteins) in extracellular T. gondii through the effects of brefeldin A (BFA). Ultrastructural studies of BFA-treated parasites showed disassembly of the Golgi apparatus and accumulation of GRA proteins in a dilated vacuolar system connected to the nuclear envelope. BFA reversibly blocked the intracellular transport of the newly synthesized GRA proteins in a dose-dependent manner (blockade of 95% at 1 microg/ml of BFA). By contrast, discharge of GRA proteins from preformed dense granules was unaffected by BFA over a course of 60 min incubation. GRA protein secretion was dependent on incubation temperature as it only occurred above 26 degrees C and it could be stimulated by external factors. This stimulus might be provided by factor(s) present in the serum of the extracellular medium, as incubation of parasites in serum-free medium resulted in a dramatic decrease in protein secretion. Exocytosis can be restored in a dose-dependent fashion by serum addition (maximal stimulatory activity in the 30-200 kDa range) and was optimal at an extracellular pH of 6.5. Altogether, these results demonstrate that GRA proteins are exported through the Golgi apparatus via the classical secretory pathway and can be experimentally discharged from storage dense granules as regulated secretory proteins in response to specific stimulation, arguing in favor of a regulated component for dense granule exocytosis in T. gondii. PMID- 10472798 TI - A homologue of Sar1p localises to a novel trafficking pathway in malaria-infected erythrocytes. AB - We have identified a homologue of the GTP-binding protein, Sar1p, in Plasmodium falciparum. Sar1p is a small GTPase that is thought to play a crucial role in trafficking of proteins between the endoplasmic reticulum and the Golgi. The P.falciparum SAR1 gene is located on chromosome 4 and comprises two exons separated by a 508 bp intron. The deduced amino acid sequence of PfSar1p (GenBank accession number AF104306) shows 71% similarity (58% identity) to Sar1p from Saccharomyces cerevisiae. Expression of PfSar1p in erythrocytic stages of P. falciparum was confirmed by sequencing of a tryptic peptide derived from a polypeptide excised from an SDS-polyacrylamide gel. A recombinant protein corresponding to approximately 70% of the PfSar1p sequence was used to raise antibodies. The affinity-purified antiserum recognised a protein with an apparent molecular weight of 23 K in Western blots of malaria-infected erythrocytes but not in uninfected erythrocytes. PfSar1p was shown to be largely insoluble in non ionic detergent and a low ionic strength buffer. Confocal immunofluorescence microscopy of malaria-infected erythrocytes was used to show that PfSar1p is located near the periphery of the parasite in discrete compartments, which appear to be distinct from the parasite endoplasmic reticulum. In addition, PfSar1p appears to be exported to structures outside the parasite in the erythrocyte cytoplasm. The export of PfSar1p to the erythrocyte cytosol is inhibited by treatment with brefeldin A. This provides the first evidence that the malaria parasite is capable of elaborating components of the classical vesicle-mediated trafficking machinery outside the boundaries of its own plasma membrane. PMID- 10472800 TI - Rapid reduction of MDCK cell cholesterol by methyl-beta-cyclodextrin alters steady state transepithelial electrical resistance. AB - The role of plasma membrane lipids in regulating the passage of ions and other solutes through the paracellular pathway remains controversial. In this study we explore the contribution of cholesterol (CH) in maintaining the barrier function of an epithelial cell line using the CH-solubilizing agent methyl beta cyclodextrin (MBCD) to stimulate CH efflux. Inclusion of 20 mM MBCD in both apical and basolateral media reduced CH levels by 70-80% with no significant effect on cell viability. Most of that decrease occurred during the first 30 min of incubation. Recovery of CH content to initial values was nearly complete 22 h after removal of MBCD. Within 30 min of adding MBCD to the culture medium, transepithelial electrical resistance (TER) increased, reaching maximum values 30 40% above controls. This early rise in TER occurred when MBCD was added to either side of the monolayer. The later rapid decline in TER was observed only when MBCD bathed the basolateral surface from which, coincidentally, CH efflux was most rapid. Freeze fracture replicas and transmission electron microscopy of monolayers exposed to MBCD for only 30 min revealed no increase in either the average tight junction (TJ) strand number or the dimensions of the lateral intercellular space. There was a statistically significant increase in the number of TJ particles associated with the E fracture face at this time. This raises the interesting possibility that during CH efflux there is a change in the interaction between TJ particles and underlying cytoskeletal elements. There was no change in staining for occludin and ZO-1. After exposing the basolateral surface to MBCD for 2 h, TER fell below control levels. The accompanying increase in mannitol flux suggests strongly that the decrease in TER resulted from an increase in the permeability of the paracellular and not the transcellular pathway. A decrease in immuno-staining for occludin and ZO-1 at TJs, a striking accumulation of actin at tri-cellular areas as well as a decline in the number of parallel strands, as seen in freeze fracture replicas, suggest that changes in cytoskeletal organization during long incubations with MBCD had physically disrupted the TJ network. Data are presented which suggest that the observed changes in paracellular permeability during CH efflux may be related to increased levels of lipid-derived second messengers, some of which may trigger changes in the phosphorylation status of TJ proteins. PMID- 10472801 TI - Beta1 integrin antisense oligodeoxynucleotides: utility in controlling osteoclast function. AB - The involvement of beta1 integrins in osteoclast function has been investigated by utilising an antisense oligodeoxynucleotide (ODN) approach. 18-mer antisense and control phosphorothioate ODNs were made to a conserved internal region of beta1 integrin sequence (nucleotide positions 1634-1651 of the human beta1 fibronectin receptor). These were tested on rabbit osteoclasts for anti-adhesive and resorptive effects mediated by alphaVbeta3 and alpha2beta1, the major integrins of osteoclasts. Antisense, but not control, beta1 ODNs inhibited osteoclast adhesion to collagen-coated glass (by up to 70%), but not to glass coated with vitronectin, fibronectin or fibrinogen. Adhesion to dentine and subsequent resorption were also inhibited (up to 60%) in a sequence-specific manner. The mechanism of action was verified using both a melanoma cell line, DX3, which expresses multiple integrins at high level including alphaVbeta3 and alpha2beta1, and in a rabbit osteoclast marrow culture (BMC) system. Exposure of DX3 cells to antisense ODN for up to 48 hours reduced adhesion to FCS- and collagen-coated glass, and concomitantly inhibited beta1 protein expression assessed by FACS and Western blot analysis; expression of other integrin subunits, alphaV and beta3, was unaffected. Similarly, the beta1 protein levels in the BMC were reduced by > 75% without any effect on actin expression. These data reveal the utility of antisense ODNs in exploring osteoclast biology and further define the functional role of osteoclastic beta1 integrin(s). PMID- 10472802 TI - Alterations in the expression and localization of protein kinase C isoforms during mammary gland differentiation. AB - Protein kinase C (PKC) is involved in signaling that modulates the proliferation and differentiation of many cell types, including mammary epithelial cells. In addition, changes in PKC expression or activity have been observed during mammary carcinogenesis. In order to examine the involvement of specific PKC isoforms during normal mammary gland development, the expression and localization of PKCs alpha, delta, epsilon and zeta were examined during puberty, pregnancy, lactation, and involution. By immunoblot analysis, expression of PKC alpha, delta, epsilon and zeta proteins was increased in mammary epithelial organoids during the transition from puberty to pregnancy. In mammary gland frozen sections, PKCs alpha, delta, epsilon and zeta were stained in the luminal epithelium and myoepithelium, in varying isoform-and developmental stage-specific locations. PKC alpha was found in a punctate apical localization in the luminal epithelium during pregnancy. During lactation, PKC epsilon was present in the nucleus, and PKC zeta was concentrated in the subapical region of the luminal epithelium. Additionally, marked staining for PKCs alpha, delta, epsilon, and zeta was observed in the myoepithelial cells at the base of ducts and alveoli. This basal ductal and alveolar staining differed in intensity in a developmentally-specific fashion. During most time points (virgin, pregnant, lactating, and early involution), myoepithelial cells of the duct were more intensely stained than those lining the alveoli for PKCs alpha, delta, epsilon and zeta. During late involution (days 9-12), the preferential staining of ducts was lost or reversed, and the myoepithelial cells lining the regressing alveolar structures stained equally (PKCs epsilon and zeta) or more intensely (PKCs alpha and delta), coincident with the thickening of the myoepithelial cells surrounding the regressing alveoli. The increased PKC isoform staining at the base of alveoli during involution suggests that alveolar regression may be influenced by alterations in signaling in the alveolar myoepithelium. PMID- 10472803 TI - Selective changes in EGF receptor expression and function during the proliferation, differentiation and apoptosis of mammary epithelial cells. AB - Epidermal growth factor (EGF) is a multifunctional regulator of mammary epithelial cells (MEC) that transduces its signals through the EGF receptor (EGFR). To clarify the role of the EGFR in the mammary gland, EGFR expression, localization and function were examined during different developmental stages in rats. Immunoblot analysis demonstrated high levels of EGFR during puberty, pregnancy and involution as well as at sexual maturity, and low levels throughout lactation. An immunohistochemical assay was used to show that EGFR was distinctly expressed in a variety of cell types throughout mammary glands from virgin rats and rats during pregnancy and involution, and was down-regulated in all cell types throughout lactation. To examine the relationship between EGFR expression and function, primary MEC were cultured under conditions that induced physiologically relevant growth, morphogenesis and lactogenesis. Cultured MEC expressed an in vivo-like profile of EGFR. EGFR was high in immature MEC, down regulated in functionally differentiated MEC, and then up-regulated in terminally differentiated and apoptotic MEC. An inhibitor of the tyrosine kinase domain of EGFR was used to demonstrate that EGFR signaling was required for growth and differentiation of immature MEC, and for survival of terminally differentiated MEC, but not for maintaining functional differentiation. PMID- 10472804 TI - Suppression of the interleukin-2 gene expression by aflatoxin B1 is mediated through the down-regulation of the NF-AT and AP-1 transcription factors. AB - The effect of aflatoxin B1 (AFB1) on the interleukin-2 (IL-2) gene expression was investigated in thymocytes of B6C3F1 mice, Jurkat E6-1 human T-cell leukemia, and EL4.IL-2 murine thymoma. AFB1 inhibited the phorbol-12myristate-13 acetate/i6nomycin (PMA/Io)-induced IL-2 mRNA expression in the murine thymocytes and Jurkat E6-1 cells as determined by qualitative RT-PCR, while no effect was observed in the EL4.IL-2 cells. Electrophoretic mobility shift assay indicated that AFB1 treatment showed an inhibition of the NF-AT and AP-1 DNA binding in PMA/Io-stimulated thymocytes and Jurkat E6-1 cells. No effect was observed on the Oct and NF-kappaB DNA binding. Employing a reporter gene expression system with p(NF-AT)3-CAT and p(AP-1)3-CAT, treatment with AFB1 to the transfected Jurkat E6 1 cells also showed an inhibition of the PMA/Io-induced NF-AT/CAT and AP-1/CAT activities. These results suggest that suppression of the IL-2 gene expression by AFB1 is mediated through the down-regulation of the NF-AT and AP-1 activation. PMID- 10472805 TI - Daily fluctuation of 7-alkoxycoumarin O-dealkylase activities in the liver of male F344 rats under ad libitum-feeding or fasting condition. AB - It has been reported that drug metabolizing enzyme activities in the liver fluctuate daily, though the daily fluctuation of 7-alkoxycoumarin O-dealkylase (ACD) activities catalyzed by P450 has not been examined. The ACD activities are known to be useful to clarify the extensive induction of P450-dependent oxygenases. In the present study, the hepatic ACD activities, as well as P450 content, were investigated periodically in male F344 rats under ad libitum feeding or fasting condition. The ACD activities in ad libitum-fed rats were found to follow obvious daily fluctuations with high values during dark periods and with low values during light periods. This agreed with the other previous results of drug metabolizing enzyme activities measured with other substrates. Because the daily fluctuation was also observed in fasted rats, the fluctuation in ACD activities was clearly not affected by feeding, an external factor. P450 content, however, showed different fluctuation patterns from the ACD activities. This suggests that the fluctuation of ACD activities might not be related to the quantitative variation of P450 proteins. PMID- 10472806 TI - Adverse effects of diphenyltin dichloride on initiation and maintenance of pregnancy in rats. AB - The objective of this study was to characterize the adverse effects of diphenyltin dichloride (DPTCl) during early pregnancy. Following successful mating, female rats were given DPTCl by gastric intubation at 0, 4.1, 8.3, 16.5, 24.8 or 33.0 mg/kg on days 0-3 or days 4-7 of pregnancy. Female rats were sacrificed on day 20 of pregnancy and pregnancy outcome was determined. The pregnancy rate was significantly decreased after administration of DPTCl on days 0-3 at 24.8 mg/kg and on days 4-7 at 33.0 mg/kg. The incidence of preimplantation loss was significantly increased after administration on days 0-3 at 16.5 mg/kg and above and on days 4-7 at 33.0 mg/kg. In females having implantations, the numbers of implantations and live fetuses and the incidences of pre- and postimplantation loss in the groups given DPTCl on days 0-3 were comparable to the controls. The incidence of postimplantation loss was significantly increased after administration of DPTCl on days 4-7 at 33.0 mg/kg. A pair-feeding study revealed no evidence of pre- and postimplantation embryolethality induced by food restriction. It could be concluded that DPTCl during early pregnancy causes early embryonic loss and DPTCl has greater effects on reproduction when administered during earlier than later stages of blastogenesis. PMID- 10472807 TI - In vitro and in vivo toxicological study of the Pterodon pubescens seed oil. AB - The oil of Pterodon pubescens seeds (PpSO) is known for its cercaricidal and anti inflammatory effects. Its anti-rheumatic activity was recently reported using mice with collagen II-induced arthritis treated with a hydroalcoholic extract of PpSO, mimicking the wine infusion used in popular medicine. In the present study, PpSO was tested for acute toxicity, mutagenic activity and cytotoxicity for human peripheral blood mononuclear cells (PBMNC). PpSO was obtained after seed extraction with 100% ethanol and evaporation. Cytotoxicity was estimated using the tetrazolium salt reduction test (MTT assay) by PBMNC (2.5 x 10(5) cells/ml) after exposure to 0.07, 0.7 and 7 microg PpSO/ml for 24 and 48 h. In the mutagenesis assay, the Salmonella/mammalian microsome assay was employed with or without metabolization. Acute toxicity was studied on 30 (n = 10/group) male DBA1/J mice (20 +/- 2 g) after a single oral dose of 2, 4, and 8 g PpSO/kg b.w. The animals were observed for 24 h, anesthetized, sacrificed and autopsied. The organs were processed for histopathology by staining with hematoxylin-eosin. The IC50 of PpSO to PBMNC in RPMI 1640 supplemented with 5% fetal calf serum (FCS) was 2 and 1 microg PpSO/ml after 24 and 48 h, respectively. The mutagenic test performed with or without metabolic activation of PpSO did not show mutagenic activity for the concentrations tested (7 and 70 microg/ml). Mouse mortality or significant signs of acute toxicity (ocular, cardiovascular, gastrointestinal, motor or respiratory signs) for the PpSO doses tested was not observed. The organs did not show any macroscopic alterations. Histopathologic analysis of the tissues also did not demonstrate any lesions. The present study provides data to classify PpSO as non-cytotoxic to PBMNC, non-mutagenic, and non-toxic after acute administration since the PpSO doses tested were extremely higher than those used by the population. PMID- 10472808 TI - Activation of the promoters of genes associated with DNA damage, oxidative stress, ER stress and protein malfolding by the bile salt, deoxycholate. AB - Toxic bile salts, retained within the liver because of impaired biliary excretion, are considered to play a major role in liver injury during cholestasis. Bile salts cause cellular stresses that may result in apoptosis. To better understand such cellular stresses, the effect of the bile salt sodium deoxycholate (NaDOC) on activation of 13 specific gene promoters or response elements associated with different cellular stresses was measured in the transformed human hepatoma line, HepG2. NaDOC was found to activate transcription factors and induce or activate the promoters of genes that respond to protein malfolding (grp78 and hsp70), DNA damage (gadd153, hsp70 and c-fos), oxidative stress (NF-kappaB, c-fos, hsp70 and gadd153), ER stress (grp78) and Ca++ imbalance (grp78). PMID- 10472809 TI - Absence of synergistic effects on micronucleus formation after exposure to electromagnetic fields and asbestos fibers in vitro. AB - Exposure of human amniotic fluid (AFC) cells to horizontally applied magnetic fields (hMF) of 50 Hz and 1 mT generated in a Helmholtz-coil system leads to a significant increase in micronucleus frequency (MN), without affecting cell proliferation. To investigate whether hMF-exposure has an additive or synergistic effect on the genotoxic capacity of asbestos fibers, MN induction was investigated in hMF pre-exposed cells, treated before or after with asbestos (1 microg/cm2). Neither synergistic nor additive effects on MN induction were observed. The results indicate, that under our experimental conditions, exposure to hMF and treatment with asbestos fibers possess genotoxic capability, but no interactive effects, in AFC cells. PMID- 10472810 TI - Elevation of metallothionein level in preneoplastic lesions during chemical hepatocarcinogenesis of the Fischer 344 rat. AB - Metallothionein (MT) is inducible by various stimuli such as metals and physiological stress. Although elevated MT expression in certain type of tumors has been reported, there are few data available on the roles of MT in tumor formation. In this study, we immunohistochemically examined MT expression in gluthathione S-transferase placental type p (GST-P)-positive preneoplastic liver lesions induced by a chemical carcinogen and further examined the relationship between MT expression and the proliferative activity of the preneoplastic cells, using bromodeoxyuridine (BrdU) labeling indices. Eleven male Fischer rats (F344/DuCrj, 9 weeks old) were subjected to the Solt-Farber protocol. Thereafter, the livers were removed, frozen and sectioned serially for immunohistochemical staining of MT and GST-P. Eight rats were given six injections of BrdU before sacrifice. In 92.8% of GST-P positive lesions, high MT staining intensity was demonstrated. On the other hand, there was no correlation between the intensity of MT staining and the BrdU labeling indices of the preneoplastic lesions. Our results indicate that MT is a useful positive marker for preneoplastic liver lesions, though the reason why MT is expressed in the preneoplastic and neoplastic lesions remains to be elucidated. PMID- 10472811 TI - The detection and comparison of the genotoxic effects of some nitro aromatic compounds by the umu and SOS chromotest systems. AB - Four nitroarenes were tested in two standard genotoxicity assay systems using three well-known bacterial tester strains. The results were as follows: 4 nitroquinoline l-oxide (4NQO) was positive in Quillardet and Hofnung's SOS chromotest using Escherichia coli strain PQ37 both in the presence and absence of microsomal (S9) supplements and in the Salmonella typhimurium umu tester strains NM2009 and NM3009, which express high levels of O-acetyltransferase (O-AT) and O AT plus nitroreductase (NR) respectively. m-Nitrocinnamic acid (m-NCA) was weakly positive in strains NM2009 and NM3009, but negative in the SOS chromotest; m dinitrobenzene (m-DNB) was weakly positive in strain NM2009, intermediate positive in strain NM3009, but negative in the SOS chromotest; 2,4-dinitrotoluene (2,4-DNT) was weakly positive in strain NM3009, but negative in strain NM2009 and in the SOS chromotest. However it still showed a dose-response relationship in strain NM2009. In view of these results, it is suggested that investigators planning to screen miscellaneous nitroarenes for their genotoxicity in the future should consider taking advantage of the increased sensitivity which is conferred on S. typhimurium strains NM2009 and NM3009 by virtue of their capacity to overexpress O-AT or O-AT and NR. PMID- 10472812 TI - Commodifying the polyvalent good of health care. AB - This essay serves as an introduction to this issue of the Journal of Medicine and Philosophy on commodification and health care. The essay attempts to sharpen the articulation of generally expressed worries about the commodification of health care. It does so by defining commodification, analyzing three components of the good of health care, and attempting to assess how commodification might distort the shape of each of those components. Next, it explores how the good of health care might be distorted by the market-based principle of distributive justice, "to each according to ability to pay." Finally, it identifies two basic questions about the relationship of medicine and the market that merit further exploration. (1) How does the market-based language of "incentives" so pervasive in the world of managed care distort the complex patterns of virtue and vice that motivate actors in the health care arena? (2) If we recognize that we cannot eliminate the influence of money from the health care system, how can we insure that the good of health care remains, in Radin's terms, "incompletely commodified"? PMID- 10472813 TI - Medicine and the market: a research agenda. AB - One of the most important developments in international medicine over the past two decades has been a turn to the market as a way of coping with rising costs and responding to calls for more freedom from government control. A full moral evaluation of the relationship of medicine and the market requires asking a wide range of questions bearing on the meaning and impact of market strategies on the economics of health care and on the clinical and public health outcomes of those strategies. A number of the leading questions are presented and some provisional answers offered. PMID- 10472814 TI - The commodification of medical and health care: the moral consequences of a paradigm shift from a professional to a market ethic. AB - Commodification of health care is a central tenet of managed care as it functions in the United States. As a result, price, cost, quality, availability, and distribution of health care are increasingly left to the workings of the competitive marketplace. This essay examines the conceptual, ethical, and practical implications of commodification, particularly as it affects the healing relationship between health professionals and their patients. It concludes that health care is not a commodity, that treating it as such is deleterious to the ethics of patient care, and that health is a human good that a good society has an obligation to protect from the market ethos. PMID- 10472815 TI - Biotechnology and commodification within health care. AB - The biotechnology industry's intellectual property claims contribute to a subtle but not insignificant encroachment of commodification within health care. Drawing on the conceptual framework of Margaret Jane Radin, I argue that patent claims on human biological materials may commodify that with which our personhood and individuality is intertwined but that such commodification is broad and incomplete. Patents on nonhuman biological organisms contribute to a more materialistic understanding of them but do not significantly change our relationship to them. The systemic effects of biotechnology's commodification within health care are various and may compromise the goal of good health. The morally problematic aspects of patent claims entail certain obligations to inhibit commodification from becoming more egregious, but on balance, those aspects are currently insufficient to justify denying the benefits the patent system promotes. PMID- 10472816 TI - Selling bits and pieces of humans to make babies: The gift of the magi revisited. AB - Reproductive medicine, a sector of a health care system increasingly captured by the demands of the marketplace, is enmeshed in a drive to sell certain human bits and pieces, such as gametes, cells, fetal eggs, and fetal ovaries, for reproductive purposes. The ethical objection raised by Kant and Radin to the sale of human organs - that this is incompatible with human dignity and worth - also applies to these sales. Moreover, such sales nullify the reproductive paradigm, irretrievably replacing it with a manufacturing paradigm. This represents a change in kind, not just of degree, in the way that we view our capacity to generate children and destroys our concept of reproduction as an essentially human activity. In the face of a struggle to retain those common ethical values at the foundation of reproductive medicine, this form of commodification of the human body should be viewed as ethically unacceptable. PMID- 10472817 TI - More questions than answers: the commodification of health care. AB - The changing world of health care finance has led to a paradigm shift in health care with health care being viewed more and more as a commodity. Many have argued that such a paradigm shift is incompatible with the very nature of medicine and health care. But such arguments raise more questions than they answer. There are important assumptions about basic concepts of health care and markets that frame such arguments. PMID- 10472818 TI - Electroacupuncture reduces rat's neuronal ischemic injury and enhances the expression of basic fibroblast growth factor. AB - The present study was designed to investigate whether the Electroacupuncture (EA) is beneficial to extenuate cerebral injuries following transient Middle Cerebral Artery Occlusion (MCAO), as well as to observe the effect of EA on expression of Basic Fibroblast Growth Factor (bFGF) -like Immunoreactivity (IR) in rat brains. The results indicate that gross neuronal damages include infarction, swelling and neuron loss, accompanied by increased bFGF-like IR expression following MCAO. In peri-infarct striatum, bFGF-like IR was mainly located in astrocytes except some neurons also showed an upregulation of the IR; in frontoparietal cortex, strong induction of bFGF-like IR was mostly seen in neurons. Both the EA applied during ischemia and reperfusion could evidently alleviate cerebral lesion extent, notably upregulate the expression of bFGF-like IR in striatum and cortex, but there was no significant difference between the effects of EA applied during ischemia and reperfusion, except EA applied during reperfusion seems to be more effective in reducing the cerebral swelling. The results implied that, in striatum, astrocytes might play an important role in the protection of neuron via the expression of bFGF; whereas in cortex, neurons may exert autoprotection through secreting bFGF themselves. One possible protective effect of EA lies in regulating the endogenous expression of bFGF. PMID- 10472819 TI - Effect of Kampo formulations (traditional Chinese medicine) on circulatory parameters. AB - The pharmacological action of 6 main Kampo formulations (1.Mao -to: [Japanese pictograph see text] MA HUANG TANG; 2. Shimbu -to: [Japanese pictograph see text]: ZHEN WU TANG; 3. Ninjin -to: [Japanese pictograph see text] : REN SHEN TANG; 4.Shigyaku-san: [Japanese pictograph see text] : SI NI SAN; 5.Keishi-to: Japanese pictograph see text] : GUI ZHI TANG; 6. Shimotsu - to: [Japanese pictograph see text] : SI WU TANG) on circulatory and autonomic nervous system were studied. 7 healthy adult males( age, 22.3 +/- 1.8 years old ) had 6 basic Kampo formulations, followed by noninvasive measurement of systolic blood pressure (SBP), mean blood pressure (MBP), diastolic blood pressure (DBP), heart rate (HR), stroke volume (SV), cardiac output (CO ), cardiacindex (CI), total peripheral resistance (TPR) by means of systolic area method of brachialsphygmography, every 30 minutes for 2 hours. As results, Mao - to induced an increase of BP,HR,SV,CO and CI, but a decrease of TPR. Keishi - to induced an increase of SBP and SV, and Shimotsu-to induced an increase of DBP and MBP, HR was slowed during former period after oral administration of Shigyaku - san, and later period after oral administration of Shimbu-to and Shimotsu-to. Regarding autonomic activity, Mao-to(former period of experiment ), Shimbu - to and Shimotsu-to induced supression of sympathetic activity, on the other hand, Mao-to (later period of experiment ) and Shiyaku - san showed a tendency of parasympathomimetic action. Mao -to induced the strongest activation of circulatory system of 6 main farmulations, and showed change of autonomic nervous activity, however, the change of circulatory and automonic nervous activity were not coincident each other. It was speculated that comprehensive mechanism of Mao to were not only dependent of ephedrin, main active constituent of Mao, but also dependent on Keishi's vasodilatory action in it. Ninjin -to showed no actions on circulatory or autonomic system. This is indicated that there are difference of actions on circulatory and autonomic nervous system among 6 main Kampo fromulations. PMID- 10472820 TI - Instrumental measurements of different homeopathic dilutions of potassium iodide in water. AB - Although more than 200 years have elapsed since the beginning of homeopathy and in spite of numerous confirmatory scientific experiments, the so-called memory of water is still a highly disputable and controversial theme in scientific circles. To make a contribution to solving this riddle, our research group tried to examine memory properties of water by the method of differential corona discharge Kirlian electrophotography of water-drop pairs. The method is based on a modified form of Kirlian photography with a subsequent thorough computer picture analysis. The potassium iodide (KI) mother solution (0.1M) was diluted in the standard way (without potentisation) or with potentisation (succussion by hand - by striking the vial 60 times against a large book as used traditionally) to 10(-3)M, 10( 6)M, 10(-10)M, 10(-16)M, 10(-17)M and 10(-24)M KI solutions. In the electrophotography method a drop of KI solution was compared with a drop of control water. To get a dependable system of results we compared homeopathic dilutions with ordinary distilled water, sham-potentised distilled water and non potentised (standard) solutions. The results were analyzed by the Chi-square Goodness-of-fit test and the Sign test. They showed repeatable and statistically significant effects of concentration of KI dilutions as well as potentisation on the corona discharge process (from p < 0.05 to p < 0.001). This indicates that there is some physical basis of molecular (ionic) information imprinted into water. PMID- 10472821 TI - Acupuncture inhibits the decrease in brain catecholamine contents and the impairment of passive avoidance task in ovariectomized mice. AB - The effects of acupuncture on the disorders elicited by abnormalities of endocrine system were investigated in ovariectomized mice. Female mice (strain; C57BL/6) were ovariectomized (OVX) and acupuncture points, Shenshu ([Japanese pictograph see text] : BL23) on both side of the back were continuously stimulated by subcutaneous needles for 20 days. After completion of experimental sessions, animals were sacrificed and specific brain regions were assayed for catecholamine contents by high performance liquid chromatography with electro chemical detector (ECD-HPLC). The mitogenic activities of splenic lymphocytes were measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTS) assay and alkaline phosphatase (ALP) assay. Furthermore, the effects of needle stimulation on learning and memory ability were studied by the step-through type passive avoidance test. Norepinephrine and dopamine contents in the frontoparietal cerebral cortex, ventral hippocampus and olfactory bulb were decreased in the OVX group, and both MTS activity and ALP activity were decreased 20 days after ovariectomy. The mean latent period was also shortened in the passive avoidance test in the OVX group. However, applying needle stimulation increased norepinephrine and dopamine contents in the brain regions, and enhanced mitogenic activities of splenic lymphocytes. The stimulation also improved memory related behavior. It was concluded from this study that after mice were stimulated by subcutaneous needle insertion, overall changes were observed in central nervous system (including retention of memory) and immune functions. The study suggests that acupuncture improves the memory loss and decrease of immune responses accompanying aging and/or menopause, and the that it may have an important role in medical care for the elderly. PMID- 10472822 TI - Encounter with my life saver, Bi-Digital O-Ring Test. PMID- 10472823 TI - Knee manipulation after total knee arthroplasty. AB - To determine if any factors are associated with knee stiffness after total knee arthroplasty (TKA), we retrospectively reviewed the medical records and radiographs of patients who had knee manipulation after total knee replacement at Scott & White Memorial Hospital from 1983 to 1993. Twenty-five patients who had knee manipulation after TKA were matched by surgeon, year of surgery, and age (+/ 5 years) with a study group of 25 patients who did not have knee manipulation after TKA. Patients in the manipulated group had decreased flexion at the time of discharge from the hospital after the knee arthroplasty and a decreased final flexion. The age of the patient, time from surgery to manipulation, and preoperative flexion did not correlate with final flexion attained in the manipulated group. Relative to the control study group, the manipulated group had an increase in postoperative anteroposterior femoral thickness. A decrease in patellar height was noted both in the manipulated group and in the control nonmanipulated group. There was no significant difference between groups for a change in patellar height. PMID- 10472824 TI - Unilateral elbow arthrodesis: the preferred position. AB - Twenty-five volunteers had unilateral elbow immobilization for 24 hours in each of two positions of flexion, 45 degrees and 90 degrees . Twenty-two of the 25 volunteers preferred a position of 90 degrees of flexion. Standard functional testing revealed significant limitations in each position of immobilization, confirming that there is no single optimal position of elbow arthrodesis. This study suggests that, for most individuals, 90 degrees is the preferred position of elbow arthrodesis for activities of daily living. However, factors such as age, sex, occupation, and dominance of the extremity should be considered when choosing a position of arthrodesis. PMID- 10472825 TI - Wrist arthrodesis using a wrist fusion plate. AB - All wrist arthrodeses done between 1990 and 1996 using a Synthes wrist fusion plate were reviewed. Independent assessment done by a certified hand therapist included a patient survey, standardized Jebsen-Taylor hand function test and activities of daily living test, and a Buck-Gramcko and Lohmann evaluation. We evaluated 13 wrists in 11 patients. Overall satisfaction was 100% of patients (mean follow-up, 31.5 months; range, 13 to 61 months). Mean preoperative and postoperative pain scores improved from 7.2 to 0.8, respectively, and functional scores improved from 5.0 to a postoperative mean value of 7.4. Jebsen-Taylor scores were virtually identical for fused and uninvolved wrists. There were no pseudarthroses, no plate failures, no tendon ruptures, and no significant postoperative infections; there was a single plate removal because of tenuous skin coverage. Short-term results using a comprehensive assessment of a custom plate designed for wrist arthrodesis show promising clinical outcomes. PMID- 10472826 TI - Congenital dislocation of the knee: overview of management options. AB - Congenital dislocation of the knee (CDK) is rare and includes a spectrum of hyperextension disorders of the knee. Early recognition of CDK is important, and careful evaluation is required to rule out associated hip deformity. Early manipulation, combined with splinting and casting, is the mainstay of initial treatment. Patients with seemingly fixed contractures may respond rapidly to serial casting and then can be placed in a Pavlik harness. Severe recalcitrant deformities or late presentation of the deformity may require surgical release. We highlight the importance of diagnostic categorization, show management options, and provide an overview of this rare but clinically significant problem. We present two case reports that illustrate the full range of management options. PMID- 10472827 TI - Penetrating bladder injury caused by a medially placed acetabular screw. AB - Cancellous bone screws are frequently used to improve the early stability of HA coated acetabular components during total hip arthroplasty. Avoidance of the anterosuperior and anteroinferior quadrants is recommended for transacetabular screw placement to minimize the risk of injury to intrapelvic structures. Revision arthroplasty in rheumatoid patients presents additional hazards in that the acetabular bone is often soft and deficient, and the protective depth of obturator internus and psoas is usually reduced. I report a case of delayed, but fatal, perforation of the bladder associated with a medially placed acetabular screw during revision arthroplasty in a rheumatoid patient. Unless directed safely into the superior pubic ramus, anterior quadrant screws should be avoided in these circumstances. PMID- 10472828 TI - Heterotopic ossification complicating total elbow replacement in a patient with rheumatoid arthritis. AB - Heterotopic ossification after total elbow replacement is a new complication. In this particular case, it resulted in severe limitation of motion. Excision of the heterotopic bone resulted in an excellent functional outcome for the patient. PMID- 10472830 TI - The nocebo effect: do no harm. AB - The nocebo effect creates negative expectations about symptoms and can have devastating influence on patient recovery. Just as the placebo effect works by making patients believe they will get better, the nocebo effect can serve to make patients worse. Two case histories are presented in which patients were assigned diagnoses without objective physical findings. This resulted in poor outcomes. Physicians should avoid assigning a diagnosis without objective physical evidence and thus avoid creating the nocebo effect in patients. PMID- 10472829 TI - Median nerve palsy presenting as absent elbow flexion: a result of a ruptured pectoralis major to biceps tendon transfer. AB - We describe a patient with a preexisting posttraumatic brachial plexopathy who had a complete high median nerve palsy due to rupture of the pectoralis major to biceps transfer near its distal insertion at the elbow region. PMID- 10472831 TI - Kinetics of antibody responses in hens from chicken lines divergently selected for response to sheep red blood cells. AB - Changes in antibody titers to SRBC were monitored for 180 d after inoculation in hens from two lines divergently selected for 24 generations for high (HAS) or low (LAS) antibody response to SRBC. The HAS hens not only had a higher peak of antibody response (12.9 vs 9.4), but also showed greater persistence in maintaining antibody levels than LAS hens. As a result, HAS hens exhibited higher antibody titers for the 180-d assay period than LAS hens. Antibodies to SRBC were detected in all day-old chicks hatched from HAS eggs collected 10 to 14 d after inoculation as well as 92 to 119 d after inoculation. Only a portion (20 to 75%) of progeny for LAS hens had detectable levels of antibody during the same periods. Among responders, antibody titers were higher for HAS than for LAS progeny. There was a positive correlation among antibody titers taken at different times after the inoculation with SRBC. PMID- 10472832 TI - Different commercial broiler crosses have different susceptibilities to leg weakness. AB - A trial was conducted to investigate the susceptibility of different genotypes of broilers to leg weakness. Four crosses of commercial broiler lines were assessed. Birds were reared on commercial diets at commercial stocking densities. Indices of leg weakness examined included: walking ability, tibial dyschondroplasia (TD), foot pad burn, hock burn, and angulation of the hock joint. Body weight and feed efficiency were also measured. There were small differences in BW and feed efficiency among the commercial crosses; however, there were large differences in some of the indices of leg weakness among the crosses. Three crosses had similar prevalence of TD; one cross had much less TD than the others. There were large differences in walking ability among crosses. There were also differences among crosses in the prevalence of foot pad and hock burn and angulation of the hock joint. Adjusting the observations for differences in BW did not substantially alter the findings. There were differences among genotypes regarding the correlation coefficients between walking ability and BW, walking ability and hock burn, and TD and BW. It was concluded that there were large differences in some important traits associated with leg weakness among the commercial line crosses. PMID- 10472833 TI - Whole genome scan in chickens for quantitative trait loci affecting carcass traits. AB - An experiment was conducted to enable quantitative trait loci (QTL) mapping for carcass traits. The population consisted of 10 full-sib families originating from a cross between male and female founders chosen from two different outcross broiler lines. Founder animals, parents, offspring, and grandoffspring are denoted as generation (G) 0, 1, 2, and 3 animals, respectively. Microsatellite marker genotypes were collected on G1 and G2 animals. Phenotypic observations were collected on G3 animals. Recorded traits were BW at 48 d, carcass weight, carcass percentage, breast meat color, and leg score. Average adjusted progeny trait values were calculated for each G2 animal and for each trait after adjusting phenotypic observations on G3 animals for fixed effects, covariables, the additive genetic contribution of the other parent, and differences between sexes. The average adjusted progeny trait values were used as the dependent variable in the QTL analysis. A QTL analysis was undertaken by modeling the segregation from G1 to G2, using a full-sib across family regression interval mapping approach. In total, 27 autosomal linkage groups covered with 420 markers were analyzed. Genomewise significance thresholds were derived using the permutation test and a Bonferroni correction. Two QTL, affecting two of the five analyzed traits, exceeded suggestive linkage. The most significant QTL was located on Chromosome 1 at 466 cM and showed an effect on carcass percentage. The other QTL, which affected meat color, was located on Chromosome 2 and gave a peak at 345 and 369 cM. PMID- 10472834 TI - Linkage relationships of the Z-linked silver, slow feathering, and pop-eye loci. AB - A three-point genetic linkage test was conducted to establish linear relationships of the Z-linked loci pop-eye (POP*), silver (S*), and rate of feathering (K*). Linkage values obtained in a back cross were POP* - 17.8 - S* - 2.4 - K* (n = 169). The POP* - K* distance was 19.0, supporting this linear order. The data support the previous assignment of POP* to the short arm of the Z chromosome. Based on previous reports, of the three loci studied, POP* is closest to the centromere. PMID- 10472835 TI - Restriction of feed consumption and body weight in two strains of large white turkey breeder hens. AB - Large White female-line turkey breeder hen poults (576) of two strains (N, female line and B, male line) were brooded by strain in floor pens from day of hatch until 3 wk of age (WOA). At 3 WOA, all poults were randomly distributed by strain to 48 floor pens in a curtain-sided house. Poults were feed (calorie) restricted beginning at 3 (N3, B3) or 6 (N6, B6) WOA to obtain a 45% reduction in BW at 16 WOA compared to ad libitum controls (NC, BC). At 16 WOA, all hens were gradually returned to ad libitum feeding. At 18 to 30 WOA, all hens were subjected to 8 h light/d. At 30 WOA, all hens were photostimulated with 15.5 h/d. Hens were inseminated weekly with semen from same strain, male-line breeder males. Eggs were collected daily, and all settable eggs from weeks of lay (WOL) 5 to 6, 10 to 11, 15 to 16, 20 to 21, and 24 to 25 were incubated by pen in a common incubator for each set date. Measurements included BW, feed consumption, egg production, mortality, egg fertility, egg hatchability, and poult weight. Statistical analysis of means was performed by strain. Target BW were obtained for feed restricted hens. Restricted hens increased their feed consumption upon re alimentation but did not achieve the cumulative feed consumption or BW of control fed hens by 56 WOA. Egg production of N3 hens was greater than NC hens for WOL 2 and 3. Total and settable eggs per hen were greater for N3 hens than for NC for WOL 1 to 5. There were no differences observed for egg weight, egg fertility, hatch of all eggs set, hatch of fertile eggs, or offspring weight among N hen treatments. The B0 hens produced more eggs than B3 or B6 hens. The B6 hens produced lighter weight eggs than B0 hens. It was concluded that early severe feed restriction of female line breeder hens might improve subsequent reproductive performance. PMID- 10472836 TI - The individual and combined effects of feeding moniliformin, supplied by Fusarium fujikuroi culture material, and deoxynivalenol in young turkey poults. AB - The effects of feeding diets containing either 20 mg deoxynivalenol (DON)/kg, 100 mg moniliformin (M)/kg, or a combination of DON and M (20 mg/kg DON and 100 mg M/kg) were evaluated in growing turkey poults, from 1 to 21 d of age. Feed intake and BW gains were decreased (P < 0.05) by dietary treatments containing M. Feed conversion was not affected by any of the dietary treatments, and no interactive effects on performance were evident between M and DON. Absolute weights of hearts and kidneys were increased (P < 0.05) in poults fed diets containing M. Mean cell volume was decreased by the M and DON-M treatments; however, the decrease was much smaller in poults fed the combination DON-M treatment resulting in a significant (P < 0.05) DON by M interaction. Mean cell hemoglobin and mean cell hemoglobin concentrations were not affected by any of the dietary treatments. No histological lesions were seen in control poults or poults fed DON alone. Lesions associated with dietary treatments were only observed in the heart and kidney. Poults fed diets containing M alone or the DON-M combination exhibited an increased incidence of variable sized cardiomyocyte nuclei, with numerous large giant nuclei, and a generalized loss of cardiomyocyte cross striations. Isolated renal tubules in sections of kidney were noted to have mild diffuse mineralization in poults fed M and the combination DON-M treatments. None of the response variables measured were affected by DON alone. No toxic synergy was observed when these toxins were fed simultaneously to turkey poults for 21 d. PMID- 10472837 TI - The effect of a second dexamethasone treatment on turkeys previously challenged in an experimental Escherichia coli respiratory model of turkey osteomyelitis complex. AB - In two separate experiments, turkeys that had survived immunosuppression with dexamethasone (DEX) and air sac inoculation with low numbers of Escherichia coli at 5 wk of age were maintained until 13 wk of age, at which time they were given a second treatment with DEX. All mortalities and birds that were necropsied 8 and 15 d (Experiment 1) and 21 d (Experiment 2) after the last DEX injection were scored for air sacculitis/pericarditis and turkey osteomyelitis complex (TOC). In both experiments, all of the lesions that characterize TOC were reproduced, including osteomyelitis of the proximal tibia, synovitis/tendonitis, abscesses in the soft tissues, and green liver. In Experiment 1, all mortalities after Day 7 had TOC lesions, whereas 44% of mortalities had green livers. Staphylococcus aureus was isolated from 90% of all TOC lesions cultured. In Experiment 2, the incidence of mortality, air sacculitis, TOC, and green liver as well as the heterophil:lymphocyte ratio were significantly higher in birds that had previously been treated with DEX but had never been challenged with E. coli than in birds that had survived both treatment with DEX and challenge with 25 or 50 cfu of E. coli. Staphylococcus aureus was isolated from 73% of TOC lesions cultured, whereas E. coli was isolated from only 5.4% of the lesions. These studies suggest that TOC incidence may be related to a stress-induced susceptibility to opportunistic infection. PMID- 10472838 TI - Electrical properties of the intestinal mucosa of the chicken and the effects of luminal glucose. AB - Transmural potential difference (PD), short-circuit current (Isc), and electrical resistance (R) were measured in the isolated mucosa of the duodenum, jejunum, ileum, proximal cecum, and rectum in order to characterize the electrical properties of the chicken small and large intestine. The chicken intestine was classified into three categories, regarding its electrical characteristics: 1) the duodenum, with four to five times higher R than the other segments and the lowest PD; 2) the group formed by the jejunum, the ileum, and the proximal cecum, with high PD and low R; 3) the rectum, with low PD and low R. In all segments, the addition of D-glucose into the luminal side stimulates Isc, and this effect can be reversed by phloridzin, indicating that the glucose-induced Isc increase is due to Na+-D-glucose co-transport. The effect of glucose is maximal in the rectum, with a fivefold Isc increase, suggesting that this segment may have an important role in the absorption of Na+ as well as of nutrients co-transported with Na+. PMID- 10472839 TI - Early postmolt performance of laying hens fed a low-protein corn molt diet supplemented with corn gluten meal, feather meal, methionine, and lysine. AB - Commercial White Leghorn hens (65, 63, or 70 wk of age in Experiments 1, 2, and 3, respectively) were induced molted by feed withdrawal until approximately 28% body weight was lost. All hens were then weighed, and seven replicate groups of 12 hens each were fed molt diets. In Experiment 1, three diets consisted of a corn basal diet (7.9% CP) or this diet supplemented with corn gluten meal (CGM) and Lys or feather meal (FM), Met, and Lys. In Experiments 2 and 3, varying levels of FM and FM with Met and Lys were evaluated. A 16% CP corn-soybean meal diet was used as a positive control in all experiments. The molt diets were fed for 17, 15, and 17 d in Experiments 1, 2, and 3, respectively, and production performance was measured for 8 wk from the beginning of feeding the layer diet. In all experiments, hens fed the 16% CP corn-soybean meal molt diet returned to egg production and regained body weight at a faster rate than did hens fed any of the other diets. In Experiment 1, early egg production of hens fed the corn basal diet supplemented with CGM and Lys or supplementation with FM, Met, and Lys was greater (P < 0.05) than that of hens fed the basal diet alone. In Experiment 2, very early egg production (Week 1) and body weight gain were lower (P < 0.05) for hens fed the corn basal diet than for hens fed the basal supplemented with FM, Met, and Lys. The addition of 5.75 or 8.5% FM or 5.75% FM plus Met and Lys generally increased (P < 0.05) early egg production and postmolt body weight gain compared to the corn basal diet in Experiment 3. The present study thus indicated that improved early postmolt performance may be achieved by supplementation of a low-protein corn molt diet with various combinations of CGM, FM, Met, and Lys. PMID- 10472840 TI - Transfer of dietary conjugated linoleic acid to egg yolks of chickens. AB - There is interest in increasing the conjugated linoleic acid (CLA) content of foods because of purported benefits of CLA for human health. Two experiments were conducted to determine the influence of dietary CLA concentration on CLA content of eggs. In Experiment 1, diets containing 0, 0.5, 2.5, or 5.0% CLA were fed to 26-wk-old White Leghorn hens (Hy-Line W-77) for 29 d. No CLA was detected in the yolk lipids of hens fed the control diet. Concentration of CLA in the yolk lipids linearly increased as dietary CLA increased. The maximum concentrations of CLA in the yolk lipids of hens fed 0.5, 2.5, or 5.0% CLA occurred 11 d after the start of the experiment and were 0.82, 5.82, and 11.20% of the total fatty acids, respectively. Concurrent decreases were observed in concentrations of C18:1, C18:2, C18:3, C20:4, and C22:6. Rate of egg production, body weight gain, and feed intake were not affected by dietary CLA. Average weights of eggs and yolks were decreased for hens fed 5.0% CLA compared with other dietary treatments. In Experiment 2, 62-wk-old hens were fed diets containing 0 or 5.0% CLA. Maximum CLA concentration in the yolk lipids of hens fed 5.0% CLA was less (7.43%) than that observed in Experiment 1. Feeding 5.0% CLA decreased feed intake but did not affect rate of egg production, weight of eggs, albumens, or yolks, or body weight gain through 36 d. Results of these experiments show that eggs produced by hens fed 5.0% CLA will contain 310 to 365 mg of CLA per egg. Such eggs could provide a substantial amount of CLA source in human foods. PMID- 10472841 TI - A bioassay to determine the effect of phytase on phytate phosphorus hydrolysis and total phosphorus retention of feed ingredients as determined with broilers and laying hens. AB - In order to accurately formulate diets for broilers and laying hens to meet phosphorus requirements without overfeeding, precise knowledge of an individual feed ingredient's contribution to the retainable phosphorus is needed. Seven feed ingredients, included as the sole source of phosphorus, were tested with and without the addition of 600 phytase units (FTU) phytase/kg diet, in a 5-d bioassay with 10 22-d-old male broilers. Without addition of phytase, the amounts of phytate phosphorus hydrolyzed in corn, soybean meal, wheat, wheat midds, barley, defatted rice bran, and canola were 30.8, 34.9, 30.7, 29.1, 32.2, 33.2, and 36.7%, respectively. The addition of phytase increased (P < or = 0.05) each value to 59.0, 72.4, 46.8, 52.2, 71.3, 48.0, and 55.8%, respectively. The addition of phytase increased total phosphorus retention from 34.8, 27.0, 16.0, 31.9, 40.3, 15.5, and 39.4% to 40.9, 58.0, 33.8, 43.4, 55.5, 26.5, and 45.7%, respectively. A similar bioassay was conducted with laying hens fed corn, soybean meal, and defatted rice bran. Without phytase addition, phytate phosphorus hydrolyzed in soybean meal, corn, and rice bran was determined to be 25.7, 23.0, and 36.1%, respectively, and was increased (P < or = 0.05) to 62.4, 52.0, and 50.9%, respectively, with the addition of 300 FTU phytase/kg feed. Total phosphorus retention of soybean meal, corn, and rice bran increased from 36.8, 28.6, and 35.9% to 53.4, 44.7, and 43.0%, respectively, with the addition of phytase. PMID- 10472842 TI - The effects of delayed access to feed and water on the physical and functional development of the digestive system of young turkeys. AB - Three experiments were conducted with turkeys to determine the influence of delayed access to feed and water on the development of the digestive system. In all experiments, poults were randomly assigned to three placement times, 6, 30, and 54 h posthatch. Experiments 1, 2, and 3 were terminated when poults were 10, 28, and 14 d old, respectively. In Experiment 1, six poults per treatment were sampled on Days 1, 2, 3, 4, 5, 6, 8, and 10. In Experiment 2, 12 poults per treatment were sampled on Days 1, 2, 3, 4, 5, 10, 15, and 28. The objective of Experiment 3 was to determine the effect of delayed placement on dietary MEn. Delaying access to feed and water for 54 h adversely affected BW through 10, 28, and 14 d of age in Experiments 1, 2, and 3, respectively (P < or = 0.01). Delayed access to feed and water for 54 h decreased the absolute weights of the small intestine and pancreas and reduced lengths of the small intestine through 5 d posthatch (P < or = 0.05). The relative weights of the small intestine and proventriculus were reduced by delayed access to nourishment through 4 d posthatch. Poults placed on feed at 54 h posthatch had decreased pancreatic amylase activity (P < or = 0.05) at 3 and 4 d, and trypsin activities were depressed at 3 d posthatch (P < or = 0.05). In Experiment 3, dietary MEn value determined at 4 d of age with poults placed 54 h posthatch was less (P < or = 0.07) than the MEn value obtained with poults placed at 6 h posthatch. A 54-h delay in access to feed and water generally delayed development of the digestive system, impaired nutrient utilization, and reduced BW. PMID- 10472843 TI - Studies on semduramicin and nutritional responses: 2. methionine levels. AB - Three experiments were conducted to determine whether feeding semduramicin at recommended levels would affect the broiler chicken's response to dietary methionine. In Experiment 1, three levels of methionine (0, 0.15, 0.30%) were fed to chicks in battery brooders. In Experiment 2, two levels of protein (18 and 22%) and three levels of methionine (0, 0.15, 0.30%) were fed to chicks in floor pens from 18 to 44 d. In Experiment 3, two methionine supplements were fed during each of the starter (0 and 0.21%, 0 to 18 d) and grower (0 and 0.10%, 18 to 35 d) periods to chicks in floor pens. In all experiments, male commercial broiler chicks were used, and all diets were fed with and without 25 mg/kg diet of semduramicin. The basal diets were based on corn, soybean meal, and poultry oil. In Experiment 1, there was a growth and feed conversion ratio response to methionine supplements, but there was no effect of semduramicin on growth nor any semduramicin by methionine supplement interaction. In Experiment 2, at 44 d, protein and methionine levels both influenced feed conversion ratios, but semduramicin did not. Feeding 22 vs 18% protein increased carcass and breast muscle yields and decreased abdominal fat pad weights. The results of Experiments 2 and 3 were very similar. Overall, the effects of semduramicin on growth and processing parameters were small and not significant, but considerable benefits in performance and carcass parameters could be realized by feeding the higher levels of protein or methionine. PMID- 10472844 TI - Nutrient content and protein and energy digestibilities of ethanol-extracted, low alpha-galactoside soybean meal as compared to intact soybean meal. AB - A study was conducted with adult Leghorn roosters to determine TMEn, dry matter digestibility, amino acid content and digestibility, and cellulose and hemicellulose content and digestibility of ethanol-extracted, low alpha galactoside soybean meal (SBM). Results were compared to that of commercially available 47% CP SBM. Twenty-one roosters were utilized; seven roosters were precision-fed each of the two soybean meal treatments, and seven served as feed deprived controls. Excreta were collected for 48 h after feeding. A second experiment compared the TMEn, as determined with roosters, to AMEn, as determined with male broiler chicks, of 44% and 47% CP SBM and three different ethanol extraction procedures to remove alpha-galactosides. The ethanol extraction procedure concentrated CP, gross energy, and TMEn. Dry matter digestibility of SBM increased from 52.1% (control SBM) to 63.3% (ethanol-extracted SBM) because of the ethanol extraction procedure. The average amino acid digestibility of ethanol-extracted SBM was 91.6%, compared to 88.0% for control SBM. The digestibility of methionine, alanine, valine, and lysine was improved by the ethanol extraction procedure. Total cellulose and hemicellulose digestion, measured as digestion of specific monosaccharide components, was increased by the ethanol extraction. The TMEn and AMEn values for the different control and extracted SBM were similar. These data indicate that the ethanol extraction of SBM to remove alpha-galactosides resulted in a product that is more protein and energy dense than commercial SBM. Advantages in digestion of amino acids and fiber may contribute to the increase in TMEn observed in both young and adult birds. PMID- 10472845 TI - Effect of pelleting temperature on the recovery and efficacy of a xylanase enzyme in wheat-based diets. AB - Two trials were performed to test the stability and efficacy of a commercial enzyme preparation containing xylanase and protease activities to pelleting. In Trial 1, 576 male Cobb chicks were fed wheat-based diets to 21 d with or without enzyme and pelleted after conditioning for 55 or 140 s at 70, 75, 80, 85, 90, or 95 C. In Trial 2, 2,880 male Cobb chicks were fed wheat-based diets to 42 d with no enzyme, or with enzyme addition before or after pelleting with conditioning for 30 s at 70, 80, 90, or 95 C. Enzyme addition had a positive effect on BW gain and the feed to gain ratio (FG) in Trial 1 and on FG to 42 d in Trial 2. All treatment effects were significant for intestinal viscosity in both trials. Two- (BW) and three-way (BW, FG) interactions between enzyme, temperature, and time were seen in Trial 1. With enzyme supplementation, second degree polynomial equations of performance on conditioning temperature had high R2 values for BW and FG, with temperatures between 80 and 85 C being most favorable. Enzyme activity in unsupplemented diets measured in vitro was largely eliminated at temperatures above 80 C, whereas the decline in activity in supplemented diets was linear. In spite of the decline in enzyme activity measured in vitro, intestinal viscosity of chicks fed enzyme-supplemented diets remained low with high processing temperatures, whereas that of chicks fed unsupplemented diets increased dramatically. PMID- 10472846 TI - Biochemical and molecular characterization of erthromycin-resistant avian Staphylococcus spp. isolated from chickens. AB - The epidemiology of the two common erythromycin-resistant methylase (erm) genes ermC and ermA was analyzed in 12 coagulase-negative Staphylococcus spp. and 34 coagulase-positive Staphylococcus spp. isolated from chicken. Southern hybridization indicated that only 2 of the 12 coagulase-negative Staphylococcus spp. strains contained the ermC gene on the plasmid; 1 strain of Staphylococcus xylosus harbored the ermC gene on a 2.5-kb plasmid, and 1 strain of Staphylococcus cohnii harbored the gene on a 4.0-kb plasmid. Twelve of the 34 strains of Staphylococcus aureus contained the ermC gene. Eleven of these strains had the ermC gene on a 2.5-kb plasmid, and 1 strain had the gene on a 4.0-kb plasmid. Ten of the 12 coagulase-negative Staphylococcus spp. and 22 of the 34 coagulase-positive Staphylococcus spp. harbored the ermA gene exclusively on the chromosome. Two different ermA EcoRI restriction fragment length polymorphisms (RFLP) were identified. A majority of the isolates was found to have two chromosomal inserts (8.0- and 6.2-kb EcoRI fragments) of ermA. One strain of S. aureus had different chromosomal inserts (6.4- and 5.8-kb EcoRI fragments) of ermA. Our results indicate that either the ermC or ermA gene, homologous to those described in human isolates, was present in all avian Staphylococcus spp. and that ermA was the predominant gene in coagulase-negative and coagulase-positive avian Staphylococcus spp. The size and copy numbers of the ermA gene were different from its human counterpart. PMID- 10472847 TI - Characterization of a cDNA sequence encoding the peroxisome proliferator activated receptor alpha in the chicken. AB - Peroxisome proliferator-activated receptors (PPAR) belong to the superfamily of the nuclear hormone receptors that play an important role in lipid homeostasis. A partial complementary deoxyribonucleic acid clone encoding a PPAR alpha from chicken liver was isolated and sequenced. Comparison with human, mouse, rat, and Xenopus PPAR alpha cDNA indicates a high degree of homology, especially at the level of the inferred peptide sequence (greater than 90%). The tissue pattern of expression indicates that PPAR alpha expression in the chicken is similar to that reported in other species, i.e., high expression levels in the liver, heart, and kidney, but also occurs in the lipogenic uropygial gland specific to birds. PMID- 10472848 TI - Hypothalamic neuropeptide Y mRNA is increased after feed restriction in growing broilers. AB - As part of an approach to uncover the brain mechanisms underlying the regulation of energy balance in broiler chickens, we investigated the possible role of neuropeptide Y (NPY). The NPY gene expression was measured in the hypothalamus of birds from a standard Ross male line and a Ross relaxed line. Both lines are derived from the same founder stock, but the relaxed line has not been selected for rapid growth since 1976. Birds of each line consumed feed either ad libitum or according to a standard commercial restricted feeding program. All groups of birds were killed at an average body weight of 2.4 kg. The NPY mRNA levels were significantly increased (P < 0.0001) in feed-restricted birds of both lines relative to ad libitum controls. No significant differences were detected between the lines. These results show that NPY gene expression in the broiler hypothalamus is sensitive to changes in energetic status, as it is in mammals. However, the maintenance of selection pressure for high growth rate is not associated with altered hypothalamic NPY mRNA levels in the ad libitum or restricted state. PMID- 10472849 TI - Identifying potentially subfertile toms via a sperm-binding assay. AB - We evaluated the utility of the commercial version of a new sperm-egg binding assay for detection of differences in sperm quality in samples of turkey semen from individual toms. Each sample had a swirl of 2 or more on a scale of 0 to 4. For assays conducted with fresh semen at 4 x 10(6) sperm per well, values ranged from 0.11 to 12% sperm bound to an extract of perivitelline membrane. Within-male variation averaged 0.17 percentage units, based on three ejaculates per male evaluated. Two experiments compared fertility and hatch for hens after weekly insemination with pooled semen from subpopulations of toms classified as having sperm with LOW or HIGH binding. Average fertility and hatch were lower (P < 0.05) for eggs laid by hens inseminated with semen from LOW toms in one experiment. In another experiment, hen fertility was not different between treatments after insemination during Weeks 32 to 39; however, a sharp decline in hatch was observed only for hens inseminated with semen from LOW toms after 40 wk of age. With semen from HIGH toms, hatch remained at > or = 80%. For these experiments, approximately 7% more poults were obtained from hens inseminated with semen from HIGH toms. We demonstrated that the sperm-egg binding assay detects differences in sperm quality between individuals, and these differences influence fertility. PMID- 10472850 TI - Postnatal growth of broilers in response to in ovo administration of chicken growth hormone. AB - The effect of in ovo administration of chicken growth hormone (cGH) on growth rate and efficiency of gain, organ, and long bone growth of 42-d-old broiler chickens was investigated. Eggs were injected once with 100 microL vehicle (0.03 M NaHCO3, 0.15 M NaCl, pH 8.3) per embryo or vehicle containing 100 ng cGH/100 microL per embryo (n = 630 eggs total) on one of the following Days: 1, 4, or 7 through 18 of embryogenesis. There was no significant difference in hatchability between control and cGH treatment groups on any given injection day. Cumulative feed conversion of all treatment groups was improved relative to their respective control groups (P < 0.05). In ovo administration of cGH on Day 15 or 16 of incubation increased body weights (P < 0.01) of female broilers. On the other hand, body weights of male broilers were significantly increased by treatment on Day 1 (P < 0.04). Breast weights of female broilers from treatment groups Day 15 or 16 were increased (P < 0.01, P < 0.05, respectively). Liver weights of female broilers from treatment groups Day 1 and 15 were increased (P < 0.05, P < 0.01, respectively). In contrast, in ovo administration of cGH on Day 11 of incubation increased liver weights of male broilers (P < 0.03). There was no significant difference between control and treatment groups, in terms of heart or leg weights, or in Warner-Bratzler shear force of Pectoralis profundus muscle. Hydroxyproline concentration and cross-sectional area of female broiler tibias from treatment groups Day 11 or Day 16 were increased (P < 0.05), and ultimate breaking strength (stress) of tibias from the same groups was reduced (P < 0.05). In ovo administration of cGH altered growth and tissue development of broiler chickens in a time by sex dependent fashion. PMID- 10472851 TI - Light intensity and age at first egg in pullets. AB - Layer strain pullets were reared on litter and given an 8-h photoperiod to 10 wk of age. They were then transferred to cages in which light intensity at the feed trough varied from 5 to 14 lx. Thirty birds continued on 8 h light (L):16 h dark (D) (negative controls), and another 30 birds were given 14L:10D (positive controls). Two other groups of 30 birds were given a regimen of 3 dim:8 L:3 dim:10 D with intensity during the dim phase ranging from 0.03 to 0.42 lx (very dim) or from 0.6 to 3.0 lx (marginal). Mean age at first egg (AFE) differed by 30 d between the positive and negative controls. Birds receiving very dim lighting matured 10 d earlier than the negative controls but 20 d later than positive controls. It is concluded that either the very dim light was itself nonstimulatory but had a phase shifting effect upon the biological clock that caused the 8-h normal light to fall partly in the photoinducible phase, or the first 3 h of very dim light was added to the bright phase to form an 11-h photoperiod. Birds in the top tier of the room with marginal supplementary lighting received 1.7 to 3.0 lx and matured at the same age as the positive controls, whereas those in the bottom tier received 0.6 to 0.9 lx and matured at the same age as the negative controls. Birds in the middle tier showed an intermediate AFE. It is concluded that the threshold intensity at the feed trough for white light stimulation of the photoperiodic mechanism in caged pullets lies between 0.9 and 1.7 lx. However, very dim lighting, below the threshold required for stimulation of a photoperiodic response, may shift the biological clock with unexpected consequences and, as a result, there is no known intensity of dim light that can be equated with darkness for all purposes. PMID- 10472852 TI - Comparative differences in the composition and biomechanical properties of tibiae of seven- and seventy-two-week-old male and female broiler breeder chickens. AB - Skeletal problems are common in both young and old poultry and are often related to bone weakness. They affect mortality on the farm and condemnations within processing plant and thus raise both welfare and economic concerns. To understand the basis of bone strength, the metaphyseal histology, composition, and the biomechanical properties of tibiae from 7- and 72-wk-old male and female broiler breeder chickens were compared. The biochemical constituents included ash, collagen, proteoglycan, transforming growth factor-beta (TGF-beta), pyridinium crosslinks, and the organic matrix-associated fluorescence. Blood variables were measured to determine the metabolic status of these birds as related to bone physiology. Although there were no differences in blood chemistry of 7-wk-old males and females, there were several differences between young and old birds. The levels of calcium, triglyceride, and iron were higher in older females than in older males. The collagen content was reduced, and the proteoglycan content increased in 72-wk-old hens. The TGF-beta content of bones from 7-wk-old females was higher than that of other groups of birds. Bone strength and stiffness, measured using loads at break and Young's modulus, respectively, were higher in older birds. The presence of medullary bones in 72-wk-old hens did not affect their bone strength, although it reduced strain values and increased Young's modulus. Compared to other groups, the 72-wk-old hens had a higher content of an inorganic matrix. The levels of hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) and the collagenase-extractable fluorescence of the organic matrix from older birds was higher. The decalcified bone matrix from older birds also showed higher susceptibility to bacterial collagenase than their younger counterparts. Bone strength showed positive correlations with its ash content, density, pyridinium crosslinks, and the fluorescence of the matrix. However, the correlation was strong with both pyridinium crosslinks and the fluorescence of the organic matrix. These results suggest that bone strength is influenced by the content of its collagen crosslinks. PMID- 10472854 TI - A randomized controlled trial of four doses of transdermal estradiol for preventing postmenopausal bone loss. Transdermal Estradiol Investigator Group. AB - OBJECTIVE: To determine the effects of four doses of a 7-day transdermal 17beta estradiol (E2) delivery system, including 0.025 mg/day, on bone loss in postmenopausal women. METHODS: This was a multicenter, double-masked, randomized, placebo-controlled study of the effects of transdermal E2 at doses of 0.025, 0.05, 0.06, and 0.1 mg/day for the prevention of postmenopausal osteoporosis. Efficacy was evaluated from bone mineral density of lumbar vertebrae L2-L4, radius, proximal femur, and total hip measured with dual-energy x-ray absorptiometry. Serum osteocalcin and urinary pyridinoline and deoxypyridinoline concentrations were measured. RESULTS: At 24 months, E2 doses of 0.025, 0.05, 0.06, and 0.1 mg/day resulted in mean increases in bone mineral density of the lumbar spine of 2.37%, 4.09%, 3.28%, and 4.70%, respectively, and increased bone mineral density of the total hip by 0.26%, 2.85%, 3.05%, and 2.03%, respectively. All increases were statistically significantly greater than placebo, which decreased bone mineral density by 2.49% at the spine and 2.04% at the hip. Consistent and significant improvements in biochemical markers of bone turnover also were noted at various intervals in all treatment groups. The most frequent adverse events were local reactions from the transdermal drug-delivery system, effects of estrogen, and menopausal symptoms. CONCLUSION: Transdermal E2 at doses of 0.025, 0.05, 0.06, and 0.1 mg/day effectively prevented bone loss in postmenopausal women. PMID- 10472853 TI - Uterine bleeding in postmenopausal women on continuous therapy with estradiol and norethindrone acetate. Endometrium Study Group. AB - OBJECTIVE: To investigate the incidence of uterine bleeding during 12 months of treatment with 17beta-estradiol (E2) 1 mg, unopposed or in combination with three doses of norethindrone acetate. METHODS: This study was a prospective, double masked, randomized, multicenter trial. A total of 1176 healthy postmenopausal women age 45 years and older without evidence of endometrial abnormalities were randomly assigned to receive either unopposed E2 1 mg, or continuous-combined formulations of E2 1 mg and norethindrone acetate 0.1 mg, 0.25 mg, or 0.5 mg. Any spotting or bleeding episodes during the treatment period were recorded in a daily diary and reported by weekly telephone calls. RESULTS: The incidence of bleeding was low in the combination groups, even during the initial 3 months of treatment (24-28%), after which it decreased with increasing doses of norethindrone acetate. Conversely, the incidence of bleeding increased over time with unopposed E2 1 mg. After the initial 3 months, the incidence of bleeding among the combination groups was lowest in the norethindrone acetate 0.5 mg group. Among women initiating therapy close to menopause, fewer reported bleeding with norethindrone acetate 0.5 mg than with the other combination groups. There was a significantly (P<.05) lower discontinuation rate due to bleeding in the norethindrone acetate 0.5 mg group compared with all other treatment groups. CONCLUSION: Continuous-combined formulations of E2 1 mg with norethindrone acetate 0.1, 0.25, or 0.5 mg are associated with a low incidence of uterine bleeding. After the initial 3 months of treatment, bleeding profiles improved with increasing doses of norethindrone acetate. PMID- 10472855 TI - Serum androgen levels and muscle mass in women with polycystic ovary syndrome. AB - OBJECTIVE: To relate serum androgen levels and muscle size in women with polycystic ovary syndrome (PCOS). METHODS: Forty-eight women with PCOS were studied. Baseline characteristics included age, height, weight, and hirsutism. Total-body lean mass was measured by wholebody scanning with dual-energy x-ray absorptiometry, and the lean mass index (total-body lean mass/height2, kg/m2) was calculated. Trunk-leg fat mass ratio (trunk-leg fat ratio) was assessed by dual energy x-ray absorptiometry. Serum testosterone, dehydroepiandrosterone sulfate, and androstenedione levels were measured by radioimmunoassay. Subjects were arbitrarily divided into two groups according to lean mass index (lean mass index at least 14 kg/m2, n = 24; or less than 14 kg/m2, n = 24). Baseline characteristics and serum androgen levels were compared between the groups. In all subjects, lean mass index was correlated with age, height, trunk-leg fat ratio, and serum androgen levels. RESULTS: Although trunk-leg fat ratio, serum testosterone and androstenedione levels, and the prevalence of hirsutism were significantly higher in women with a lean mass index at least 14 kg/m2, age and height did not differ significantly between the groups. Trunk-leg fat ratio and serum testosterone and androstenedione levels correlated significantly with lean mass index (r = .68, .75, and .54, respectively). Hirsutism also correlated with lean mass index (standardized regression coefficient = .49; P<.05). CONCLUSION: Muscle size in women with PCOS correlated positively with serum androgen levels and upper body-fat distribution, independent of height. PMID- 10472856 TI - Hysteroscopic myomectomy: long-term effects on menstrual pattern and fertility. AB - OBJECTIVE: To determine the effects of hysteroscopic myomectomy on menorrhagia and infertility and the influence of intramural extension on surgical feasibility and long-term outcomes. METHODS: We studied 108 women who had first-line hysteroscopic resection of submucous pedunculated (n = 54), sessile (n = 30), or intramural (n = 24) leiomyomas over 7 years at an academic department specializing in endoscopic surgery. RESULTS: The mean (+/- standard deviation) operating time and distension medium deficit were 18+/-7 minutes and 204+/-276 mL in the pedunculated lesion group, 23+/-9 minutes and 278+/-269 mL in the sessile lesion group, and 32+/-8 minutes and 335+/-272 mL in the intramural lesion group, respectively. More than one procedure was required to complete myoma removal in 14 (26%) of 54, eight (26%) of 30, and 12 (50%) of 24 subjects in the pedunculated, sessile, and intramural lesion groups, respectively. After a mean follow-up of 41 months, myomas recurred in 27 subjects, with a 3-year cumulative rate of 34%. Twenty women had recurrent menorrhagia, with a 3-year cumulative probability of 30%. The 3-year cumulative probability of conception was 49% in women with pedunculated lesions, 36% in those with sessile lesions, and 33% in those with intramural lesions. The study had 80% power to detect five- and three fold increases in menorrhagia recurrence and conception rates, respectively, in the mainly intramural myoma group compared with the completely or mainly intracavitary myoma group. CONCLUSION: Hysteroscopic resection of submucous myomas gives satisfactory menorrhagia control and limited recurrence, but the benefit for infertility was less impressive. Myoma intramural extension did not have a substantial influence on any of the long-term outcomes but affected operating time and the number of procedures needed for complete removal. PMID- 10472857 TI - Vaginal removal of the benign nonprolapsed uterus: experience with 300 consecutive operations. AB - OBJECTIVE: To determine the effectiveness and safety of vaginal hysterectomy for benign nonprolapsed uteri. METHODS: Three hundred consecutive women with nonprolapsed uteri requiring hysterectomy for benign uterine conditions, without suspected adnexal disease, were treated prospectively by vaginal hysterectomy. Twenty-one women (7%) were nulliparous, and 219 (73%) had history of pelvic surgery (150 had previous cesareans). Operating time, estimated blood loss, surgical techniques (Heaney, Pelosi, uterine morcellation), operative complications, conversion to laparoscopy or laparotomy, and length of hospital stay were recorded for each case. RESULTS: Vaginal hysterectomy was successful in 297 women (99%). Morcellation (hemisection, intramyometrial coring, myomectomy, and wedge resection) was done in 170 cases (56.7%). The mean operating time was 51 minutes (range 20-130 minutes), mean estimated blood loss was 180 mL (range 50 1050 mL), and mean length of hospitalization was 22 hours (range 16-72 hours). Four operative complications occurred (three cystotomies, one rectal laceration) and were repaired transvaginally. One woman needed a blood transfusion. Eleven urinary tract infections occurred. Two conversions to laparotomy and one conversion to laparoscopy were necessary. CONCLUSION: Vaginal hysterectomy is an effective and safe procedure for benign nonprolapsed uteri irrespective of nulliparity, previous pelvic surgery, or uterine enlargement. We question the true need for laparoscopy or laparotomy in this setting. PMID- 10472858 TI - Medical and osteopathic boards' positions on chaperones during gynecologic examinations. AB - OBJECTIVE: The objective of this study was to determine whether United States medical or osteopathic boards have opinions, position statements, or policies on chaperone use, and whether any state laws regulate chaperone use. METHODS: United States Medical and Osteopathic Boards were surveyed by mail to determine whether policies, opinions, positions, or laws exist regarding use of chaperones during gynecologic examinations. We sent the survey to executives at 67 state boards, identified by a list from The Federation of State Medical Boards. Our main outcome measure was positive response to the survey questions. RESULTS: Of 67 targeted sites, 61 responded (91%). Fourteen sites (23%) reported having informal or unpublished opinions recommending chaperones. Eleven sites (18%) reported having positions related to chaperones that have been published for their physicians. Four sites (6.5%) reported having policies specifically related to chaperone use. Thirty-two sites (52.5%) reported that they do not have opinions, positions, or policies related to chaperone use. No site reported state laws governing chaperone use. CONCLUSION: Response to our survey showed no concensus among state medical boards on the use of chaperones, leaving doctors and patients to decide for themselves whether they want or need chaperones present during gynecologic examinations. PMID- 10472859 TI - Plasma homocyst(e)ine concentrations in eclamptic and preeclamptic African women postpartum. AB - OBJECTIVE: To examine the relationship between plasma homocyst(e)ine and risk of eclampsia and preeclampsia among sub-Saharan African women who delivered at Harare Maternity Hospital in Zimbabwe. METHODS: We ran a hospital-based, case control study at Harare Maternity Hospital, University of Zimbabwe, Harare, Zimbabwe comprising 33 pregnant women with eclampsia and 138 with preeclampsia. Controls were 185 normotensive pregnant women. Plasma was collected postpartum and homocyst(e)ine levels were measured by high-performance liquid chromatography and electrochemical detection. RESULTS: Women with eclampsia or preeclampsia had significantly higher mean homocyst(e)ine levels than normotensive controls (12.54 or 12.77 micromol/L versus 9.93 micromol/L, respectively, P<.001). The odds ratio (OR) for eclampsia was 6.03 among women in the highest quartile of the control homocyst(e)ine distribution (median 13.9 micromol/L) compared with women in the lowest quartile (median 6.2 micromol/L). The corresponding OR for preeclampsia was 4.57. Nulliparas with elevated homocyst(e)ine had a 12.90 times higher risk of preeclampsia compared with multiparas without elevated homocyst(e)ine. CONCLUSION: Postpartum plasma homocyst(e)ine concentrations are higher among Zimbabwean women with eclampsia and preeclampsia compared with normotensive women. PMID- 10472860 TI - Ophthalmic artery velocimetry in normotensive and preeclamptic women with or without photophobia. AB - OBJECTIVE: To compare ophthalmic arterial velocimetry in normotensive and preeclamptic gravidas with and without photophobia. METHODS: Ophthalmic arteries were studied by color-flow Doppler ultrasonography in 118 normotensive pregnant women, 20 gravidas with preeclampsia and no visual symptoms, and 11 with preeclampsia, photophobia, and retinal edema. RESULTS: The ophthalmic arterial pulsatility index (PI) correlated negatively with gestational age (y = -0.01x + 1.84, r = -0.41, P<.01). Pulsatility index in preeclamptics with photophobia (0.71+/-0.17) was lowest among the three groups (P<.01) and was highest in normotensive pregnant women (1.41+/-0.21, P<.01). Mean velocity in normotensive pregnant women (0.19+/-0.05 m/second) was highest among the groups (P<.01) and was not significantly different in preeclamptic women with no visual symptoms (0.27+/-0.03 m/second) and with photophobia (0.30+/-0.02 m/second). CONCLUSION: Preeclamptic women, especially those with photophobia, have orbital vascular vasodilation or hyperperfusion, or both. PMID- 10472861 TI - Reducing unnecessary coagulation testing in hypertensive disorders of pregnancy. AB - OBJECTIVE: To estimate the magnitude of laboratory testing for hypertension in pregnancy and determine whether abnormalities in prothrombin time (PT), activated partial thromboplastin time (aPTT), and fibrinogen can be predicted by results of common, less expensive tests. MATERIALS AND METHODS: Laboratory records were searched and charts were reviewed to identify gravidas tested for hypertension and to exclude conditions producing coagulopathy. Contingency tables were constructed to assess the ability of the platelet count, lactate dehydrogenase, and transaminases to predict coagulation test results. RESULTS: Preliminary data on 73 gravidas found that a platelet count plus a lactate dehydrogenase test best predicted coagulation abnormalities. Results on another 732 gravidas indicated that coagulation tests were obtained in about 30%. No patient had a PT greater than 18 seconds, two had an aPTT greater than 40 seconds, and three had fibrinogen levels less than 200 mg/dL. The combination of a normal platelet count plus a normal lactate dehydrogenase had a negative predictive value of 100% for clinically significant abnormalities of PT and aPTT, and 99% for significant abnormalities of fibrinogen. CONCLUSIONS: Substantial coagulation testing was done on gravidas evaluated for a hypertensive disorder even though the prevalence of clinically significant abnormalities was low. Laboratory evaluation of patients suspected of having preeclampsia need not include a PT, aPTT, or fibrinogen test when there is no evidence of bleeding or of a condition that could produce coagulopathy and when the platelet count and lactate dehydrogenase level are both normal. PMID- 10472862 TI - Predicting compliance with follow-up recommendations after colposcopy among indigent urban women. AB - OBJECTIVE: To determine correlates of compliance with follow-up recommendations after colposcopy. METHODS: Between October 1, 1992, and June 30, 1997, management recommendations were made to 451 consecutive women who attended a resident colposcopy clinic. Compliance was determined after 8-12 months. Correlates of compliance were assessed by univariate and multivariate analyses. RESULTS: Recommendations included repeat cytology in 6 months for 252 women, loop excision for 114, conization for 27, repeat colposcopy for 51, other for three, and not recorded for four. Rates of ever complying were 81% for loop excision, 85% for conization, 62% for repeat colposcopy, and 36% for repeat cytology. In multivariate analysis, the only significant predictors of compliance were history of genital herpes simplex infection (odds ratio [OR] 0.20; 95% confidence interval [CI] 0.05, 0.86, P = .03) and the nature of the recommendation (OR 4.73; CI 2.51, 8.92 for loop excision; OR 8.72; CI 2.57, 29.6 for conization; and OR 2.53; CI 1.30, 4.93 for repeat colposcopy, all against observation, P<.001). CONCLUSION: After colposcopy, women were more likely to comply with earlier, more intensive interventions, although the relative effect of follow-up interval and intervention could not be distinguished. PMID- 10472863 TI - See-and-treat in the management of high-grade squamous intraepithelial lesions of the cervix: a resource utilization analysis. AB - OBJECTIVE: To use activity-based costing techniques to compare see-and-treat with conventional evaluation and treatment of women presenting with a screening Papanicolaou smear demonstrating high-grade squamous intraepithelial lesion (SIL). METHODS: A total of 4000 theoretical patients were assumed to be evaluated and treated following one of four management algorithms: conventional algorithm I, with colposcopy and directed biopsies, followed by cryotherapy or cold-knife conization; conventional algorithm II, substituting the loop electrosurgical excision procedure for cold-knife conization; conventional algorithm III, substituting the loop electrosurgical excision procedure for cold-knife conization and cryotherapy; or see-and-treat algorithm IV, using the loop electrosurgical excision procedure. Costs associated with patient management in each algorithm were calculated including those for the procedure, patient time, physician time, and disposable expenses, as well as costs to manage complications, treatment failures, and follow-up for 1 year. RESULTS: Algorithm I was the most expensive, costing $899,405 for 1000 patients with high-grade SIL. Substituting the loop electrosurgical excision procedure for cold-knife conization (algorithm II) decreased the cost by 32%, whereas substituting it for cryotherapy also (algorithm III) reduced the cost by only 25%. The most cost effective management was the see-and-treat single visit of algorithm IV. This strategy cost $531,281, offering a 41% cost reduction compared with algorithm I. CONCLUSION: A see-and-treat approach to the management of women with high-grade SIL, although incorporating more procedures, offers significant cost savings over conventional management algorithms. PMID- 10472864 TI - Early cervical adenocarcinoma: selection criteria for radical surgery. AB - OBJECTIVE: To identify selection criteria for radical surgery in early cervical adenocarcinoma based on pretreatment clinical stage and correlation with high risk surgical-pathologic factors. METHODS: One hundred seventy-five women with International Federation of Gynecology and Obstetrics (FIGO) clinical stage IB1 (n = 132) and IB2-IIA (n = 43) cervical adenocarcinoma were treated primarily at our institutions from 1982 to 1996. Histopathologic sections were reviewed by a gynecologic pathologist. Medical records were reviewed retrospectively and clinical follow-up was done. RESULTS: The overall 5-year survival rate was 87% (95% confidence interval [CI] 81%, 93%) for stage IB1 and 61% (95% CI 46%, 77%) for stage IB2-IIA (P<.001). Adenosquamous cell type, deep cervical invasion, and lymph-vascular space invasion were significant independent high-risk surgical pathologic factors that affected disease-free survival (each P<.002). One hundred fourteen (86%) of 132 stage IB1 patients and 19 (44%) of 43 stage IB2-IIA subjects were treated primarily with radical surgery. Lymph node metastases, lymph-vascular space invasion, adenosquamous cell type, deep cervical invasion, and positive surgical margins were more than twice as frequent in stage IB2-IIA patients who had radical surgery than in stage IB1 patients (each P <.05). Based on high-risk surgical-pathologic factors in 133 subjects who had radical surgery, postoperative radiotherapy was recommended for 18 (16%) of 114 stage IB1 patients and 18 (95%) of 19 stage IB2-IIA subjects (P<.001). CONCLUSION: Radical surgery for FIGO clinical stage IB1 cervical adenocarcinoma and primary radiotherapy for stage IB2-IIA disease would largely avoid combined-modality therapy, thereby reducing treatment-related toxicity and cost. PMID- 10472865 TI - Distal external iliac lymph nodes in early cervical cancer. AB - OBJECTIVE: To evaluate the utility of the deep circumflex iliac vein as a landmark for the caudal limit of external iliac lymphadenectomy for early cervical cancer. METHODS: During dissection of the distal (caudal), anterior (ventral) aspect of the external iliac vessels in conjunction with radical hysterectomy for carcinoma of the cervix, a careful search was made for the deep circumflex iliac vein. Lymph nodes immediately above this vein were sent as adjacent and lymph nodes caudal to the vein were sent as distal. The distance from the vein to the femoral canal was measured. RESULTS: Seventy-one women were studied over 40 months. Fifty-five had squamous cell carcinoma, 15 had adenocarcinoma, and one had adenosquamous carcinoma. The mean distance from the deep circumflex iliac vein to the femoral canal was 16 mm. Sixty-three patients had dissections distal to identified deep circumflex iliac veins and 49 (77%) of these had distal lymph nodes removed. The median number of pelvic lymph nodes removed was 24 (range nine to 68), and the median number of distal lymph nodes removed was 1.0 from each side. Lymph node metastases were found in 13 women (18%). One subject with multiple macroscopically positive nodes had a single positive distal lymph node. Thus, a small number of distal lymph nodes were found in most women with early invasive cervical cancer, and 8% of those with positive nodes had involvement of this nodal group. CONCLUSION: The deep circumflex iliac vein was an appropriate landmark for the caudal limit of external iliac lymphadenectomy. PMID- 10472866 TI - Diet and uterine myomas. AB - OBJECTIVE: To analyze the relation between selected dietary indicators and the risk of uterine myomas. METHODS: We used data from a case-control study on risk factors for uterine myomas conducted in Italy between 1986 and 1997. Cases included 843 women with uterine myomas whose clinical diagnoses dated back no more than 2 years. Controls were 1557 women younger than age 55 who had not had hysterectomies and were admitted for acute nongynecologic, nonhormonal, nonneoplastic conditions. RESULTS: Women with uterine myomas reported more frequent consumption of beef, other red meat, and ham and less frequent consumption of green vegetables, fruit, and fish. The multivariate odds ratios in the upper tertile were 1.7 for beef and other red meat (95% confidence interval [CI] 1.4, 2.2), 1.3 for ham (95% CI 1.0, 1.6), 0.5 for green vegetables (95% CI 0.4, 0.6), and 0.8 for fruit consumption (95% CI 0.6, 1.0). CONCLUSION: Myoma is associated with beef and ham consumption, whereas high intake of green vegetables seems to have a protective effect. PMID- 10472867 TI - Partner notification of sexually transmitted disease in an obstetric and gynecologic setting. AB - OBJECTIVE: To survey obstetrician-gynecologists regarding current approaches to partner notification of women with sexually transmitted diseases (STDs) and to relate reported practices to state requirements. METHODS: An anonymous questionnaire was sent to community, hospital-employed, and university-based physicians within a single health care system. The survey was designed to assess physician demographics, knowledge, and attitudes about partner notification of women with STDs. RESULTS: One hundred eight (49%) of 222 surveys were returned. Respondents were 36% private practitioners, 38% hospital-employed, and 23% university faculty physicians. Although most correctly identified the importance of partner notification for several STDs, they (63 of 108, 58%, confidence interval [CI] 50%, 67%) could not identify all of those conditions. Most respondents (84 of 108, 78%, CI 69%, 85%) were unaware of all required reportable STDs and the state's preferred method of partner notification. The vast majority of respondents (96 of 108, 89%, CI 82%, 94%) relied on women to notify their partners of the STD. CONCLUSION: Educational efforts are needed to increase physician awareness of required reportable STDs and the role of partner notification and treatment in disease control. Physicians need to be aware of state regulations for partner notification when they are in practice. Development of standard algorithms for partner notification and treatment might be needed. PMID- 10472869 TI - Prognostic markers in twin pregnancies with an acardiac fetus. AB - OBJECTIVE: With twin reversed arterial perfusion sequence, the normal cotwin is at risk of heart failure. The purpose of this study was to identify ultrasonographic variables that can help predict prognosis. We hypothesized that variables reflecting the hyperdynamic circulatory condition of the normal fetus and the changes in circulatory impedance in the acardiac mass would correlate with final outcome. METHODS: Ten twin pregnancies with this condition were identified. Follow-up was available for nine. Adverse outcome was defined as death, cardiac failure, or delivery before 30 weeks' gestation for reasons related to the presence of the mass. The following data were collected on the normal fetus: cardiothoracic ratio and left ventricular shortening fraction; and on the mass: maximal length, presence and size of cysts, and presence of a rudimentary heart. The pulsatility index (PI) of the umbilical arteries (UA) of both twins was measured. RESULTS: Four fetuses died, two in utero (22 weeks) and two after cesarean (26 and 31 weeks) for advanced cardiac failure. In the five other cases, the outcome was favorable. The cardiothoracic ratio and presence of cysts or of a rudimentary heart did not correlate with outcome. A PI in the mass' UA significantly lower than that of the normal twin (ratio of 0.71 compared with 1.04 for good outcome, P<.05), an elevated shortening fraction in the second trimester, and a rapid growth rate of the mass were associated with a poor prognosis. CONCLUSION: In pregnancies with twin reversed arterial perfusion sequence, final outcome and treatment decisions can be determined based on hemodynamic criteria. PMID- 10472868 TI - Glyceryl trinitrate and ritodrine in tocolysis: an international multicenter randomized study. GTN Preterm Labour Investigation Group. AB - OBJECTIVE: To compare the efficacy of transdermal glyceryl trinitrate and intravenous (IV) ritodrine as tocolytics. METHODS: Two hundred forty-five women with preterm labor and intact membranes between 24 and 36 weeks' gestation were randomized to transdermal glyceryl trinitrate or intravenous ritodrine. Treatment was continued until contractions stopped or a maximum of 7 days. Glyceryl trinitrate was administered as a 10- or 20-mg transdermal patch. Intravenous ritodrine was administered according to nationally available guidelines. The primary outcome was prolongation of gestation expressed as a percentage of the time from entry to 37 weeks. Secondary outcomes were proportion of women who delivered the same day, next day, or within 7 and 14 days of entry, and by 32, 34, and 37 weeks. Analysis was by intention to treat. RESULTS: Twelve women (5%) were lost to follow-up. Glyceryl trinitrate and ritodrine prolonged gestation by 74% of time to 37 weeks (difference glyceryl trinitrate-ritodrine 0%; 95% confidence interval (CI) -10%, +10%). There was no significant difference in the proportion of women receiving glyceryl trinitrate or ritodrine who delivered within the specified days from study entry or weeks of gestation; however, 42 women who received glyceryl trinitrate and 58 women who received ritodrine delivered by 37 weeks (difference -11%; 95% CI -24%, +2%). No serious maternal side effects were reported for ritodrine or glyceryl trinitrate. CONCLUSION: We found no overall difference between glyceryl trinitrate and ritodrine in the acute tocolysis of preterm labor but a suggested advantage of glyceryl trinitrate over ritodrine in reducing preterm delivery rate. The maternal side effect profile and treatment discontinuation rates were fewer for glyceryl trinitrate, suggesting it was a safer alternative to ritodrine. PMID- 10472870 TI - Meconium-induced umbilical vascular necrosis in abortuses and fetuses: a histopathologic study for cytokines. AB - OBJECTIVE: To show that meconium causes fetal morbidity and death at earlier gestations than reported previously. METHODS: We searched for specimens from 1997 and 1998 with pathologic diagnosis of meconium-induced umbilical vascular necrosis in placentas of nonmalformed fetuses and newborns. Because intra amniotic infection is known to activate cytokines, and blood pigment is often microscopically indistinguishable from meconium, we removed those confounding considerations by excluding placentas with chorioamnionitis and signs of intra amniotic bleeding. Light microscopic identification of vacuolar amniotic epithelial degeneration was used to select specimens with meconium because blood does not cause that histopathologic abnormality. We used histochemical procedures to show absence of hemosiderin and presence of bilirubin, and immunocytochemical labeling with interleukin-1beta to show cytokine. RESULTS: Four cases had meconium-induced umbilical vascular necrosis. The gestational ages were 16, 19, 29, and 38 weeks. Two cases were abortuses, the third was stillborn, and the fourth was a small-for-gestational-age liveborn, delivered by cesarean because of repetitive variable decelerations. Luna-Ishak staining showed bilirubin in macrophages between umbilical vascular myocytes and in the Wharton's jelly. Immunocytochemical methods showed interleukin-1beta in those same macrophages. CONCLUSION: Cytokines and other meconium-associated factors may contribute to the pathogenesis of fetal death. Survivors may suffer intraventricular hemorrhage, periventricular leukomalacia, and other morbidity. PMID- 10472871 TI - Knowledge and clinical practice regarding calcium nutrition among obstetrician gynecologists. AB - OBJECTIVE: To survey the knowledge of and attitude toward nutrition among practicing obstetrician-gynecologists, using calcium as a specific example. METHODS: Our survey on calcium nutrition consisted of 29 questions divided into four categories: demographic information, professional interest and clinical practice, knowledge and opinions, and education. We mailed this survey to the 244 ACOG Fellows who are members of the Collaborative Ambulatory Research Network and to a random sample of 756 Fellows who are not members of the Collaborative Ambulatory Research Network. RESULTS: Most (75.4%) respondents considered making dietary recommendations an important part of their practice. Interest in nutrition was correlated significantly with its perceived importance in clinical practice and was associated with a greater self-reported likelihood of making dietary recommendations to patients and referring patients to nutritional counseling. Nutrition was of greater importance to women and to older respondents. Women were more likely to make dietary recommendations to patients. Men were more likely to recommend nonfoods and higher fat foods as sources of dietary calcium. The daily calcium intake that obstetrician-gynecologists recommended for different types of patients varied widely but did not differ between male and female respondents. The responses to the knowledge questions concerning calcium metabolism were predominantly "don't know." CONCLUSIONS: Although specific knowledge of calcium metabolism and regulation was incomplete, obstetrician-gynecologists generally recognized the importance of nutrition in their clinical practice. Being older and female independently increased the perceived importance of nutrition. PMID- 10472872 TI - A randomized controlled trial of vaginal misoprostol for cervical priming before hysteroscopy. AB - OBJECTIVE: To evaluate the effectiveness and side effects of vaginal misoprostol for cervical dilation in nonpregnant women before hysteroscopy. METHODS: Ninety one women scheduled to have hysteroscopy were randomized to receive either vaginal misoprostol or placebo. Cervical response, outcome of hysteroscopy, and side effects of vaginal misoprostol were assessed. RESULTS: The mean cervical dilatation estimated by Hegar dilator and the mean duration of hysteroscopy were significantly different between the treated group (7.0+/-1.0 mm [range 6-8.5] and 90.0+/-38.4 seconds [range 60-240], respectively) and the control group (3.8+/ 1.2 mm [range 2-5.5] and 142.0+/-38.7 seconds [range 60-270]). In the misoprostol group, only three women (6.5%) needed cervical dilation before hysteroscopy, compared with 14 (31.1%) in the placebo group (P = .006). Cervical tears during hysteroscopy occurred in two patients (4.4%) in the control group and none in the misoprostol group. The two most common side effects of vaginal misoprostol were mild lower abdominal pain in 15 women (32.6%) and slight vaginal bleeding in 12 (26.1%). Both side effects were significantly different when compared with placebo (P<.001). CONCLUSION: Vaginal misoprostol lessens the cervical resistance in women undergoing hysteroscopy and facilitates the procedure, with only mild side effects. PMID- 10472873 TI - Evacuation interval after vaginal misoprostol for preabortion cervical priming: a randomized trial. AB - OBJECTIVE: To determine the optimal interval for evacuation after preabortion cervical priming with vaginal misoprostol. METHODS: One hundred eighty healthy nulliparas requesting legal termination of pregnancy between 6 and 11 weeks' gestation were assigned randomly to receive 400, 600, or 800 microg of intravaginal misoprostol. Vacuum aspiration was done after 3 hours in the 400 microg group and after 2 hours in the 600- and 800-microg groups. The degree of cervical dilatation before operation was measured with a Hegar dilator. Preoperative and intraoperative blood loss and associated side effects were also assessed. RESULTS: Eleven (18.3%) and 15 (25.0%) women in the 600-and 800-microg groups, respectively, had cervical dilatation of at least 8 mm after an interval of 2 hours; 55 (91.7%) women who received 400 microg for a 3-hour interval had similar cervical dilatation. Using 400 microg as a baseline, the odds ratio (OR) was 0.02; 95% confidence interval (CI) was 0.01, 0.06 for 600 microg and OR 0.03; 95% CI 0.01, 0.09 for 800 microg for achieving successful preabortion cervical dilatation of at least 8 mm. The mean cervical dilatation of 6.7 mm and 6.8 mm for the higher doses was also significantly less than that of 8.1 for the 400 microg dose (P<.001). The mean preoperative and intraoperative blood loss was only statistically different when the 400- and 800-microg groups were compared (P = .03). There were also significantly more side effects, namely abdominal pain and fever above 38.0 C, in the 600- and 800-microg groups (P<.001), compared with the 400-microg group. When the 600- and 800-microg groups were compared, there were still significantly more women complaining of abdominal pain (P<.001). None of the women in the study required analgesics for pain or antipyretics for fever. CONCLUSION: The minimal evacuation interval should be at least 3 hours for successful preabortion cervical priming. PMID- 10472874 TI - Stromelysins in placental membranes and amniotic fluid with premature rupture of membranes. AB - OBJECTIVE: To determine the expression and site of production of stromelysins in fetal membranes and to measure stromelysin 1 levels in amniotic fluid and amniochorion culture media. METHODS: Amniochorionic membranes were cultured from organ explant. Membranes were stimulated with lipopolysaccharide for 24 hours after a 48-hour preincubation period. Membranes were also collected from women after vaginal deliveries. RNA samples from those tissues were subjected to reverse transcriptase-polymerase chain reaction using primers specific for stromelysin 1, stromelysin 2, stromelysin 3, and matrilysin. In situ hybridization and immunohistochemistry were used to localize stromelysin mRNA and peptide. Levels of stromelysin 1 in culture media and amniotic fluid collected from women with preterm premature rupture of membranes (PROM) and at term with intact membranes were compared using enzyme-linked immunosorbant assay. RESULTS: Amniochorion in culture and from laboring and nonlaboring women expressed all three stromelysins. In situ hybridization showed stromelysin mRNA in amnion, chorion, and extracellular matrix. Immunohistochemical analysis localized stromelysin 1 protein to those same regions. Amniotic fluid levels of stromelysin 1 were higher in preterm PROM amniotic fluids (median 3.2 ng/mL) compared with term deliveries with intact membranes (median 1.3 ng/mL) (P = .02). Lipopolysaccharide stimulation in culture increased the release of stromelysin 1 from fetal membranes compared with control (median 70.35 versus 15.8 ng/mL, respectively, P = .05). CONCLUSION: Human fetal membranes are a source of stromelysins 1, 2, and 3. Increased stromelysin 1 during preterm PROM and in vitro after lipopolysaccharide stimulation suggests a possible effect of that matrix metalloproteinase in PROM. PMID- 10472875 TI - Genes upregulated in human fetal membranes by infection or labor. AB - OBJECTIVE: To determine whether suppression subtractive hybridization can detect genes in fetal membranes that are upregulated by infection, preterm premature rupture of membranes (PROM), or labor. METHODS: Using suppression subtractive hybridization, messenger RNAs from a preterm fetal membrane obtained at cesarean delivery without labor (control) were subtracted from a pool of messenger RNAs of three patients with preterm PROM and vaginal delivery. Eight candidate genes identified as upregulated were quantitated by Northern analysis in each of the tissues and in additional patient subgroups. RESULTS: Eight differentially upregulated genes were identified in preterm labor with PROM. Four of the genes are known to be involved in the response to inflammation or infection, and subsequent histologic examination showed one of the preterm PROM tissues to be infected. F-actin capping protein and chitinase precursor, not previously known to be involved in infection, were also upregulated in the infected tissue from preterm PROM. Northern blots using additional subgroups of patients showed that a regulatory G-protein signaling protein gene was significantly upregulated at term by labor in addition to significant upregulation of interleukin-8. There was a strong correlation between the gene expression for complement factor-B and duration of membrane rupture in the patients with preterm PROM. CONCLUSION: Two novel genes potentially involved in the response to inflammation or infection have been identified. A regulatory G-protein signaling protein and interleukin-8 gene expression were upregulated by labor. Complement factor-B gene expression was directly related to the duration of membrane rupture. PMID- 10472876 TI - Cervical length at 23 weeks in twins in predicting spontaneous preterm delivery. AB - OBJECTIVE: To establish the relationship between cervical length at 23 weeks' gestation in twin pregnancies and risk of spontaneous preterm delivery. METHODS: Cervical length was measured during routine antenatal care by transvaginal sonography at 23 (range 22-24) weeks' gestation in 215 twin pregnancies. Distribution of cervical length was determined, and sensitivity and false positive rate for spontaneous preterm delivery at or before 28, 30, 32, and 34 weeks for cutoff cervical lengths of 15, 25, 35, and 45 mm were calculated. RESULTS: Cervical length distribution was skewed toward shorter length and the median value was 38 mm. In 11.2% and 4.2% of cases, length was up to 25 mm and up to 15 mm, respectively. The spontaneous delivery rates at or before 28, 30, 32, and 34 weeks were 3.8%, 4.7%, 8.0%, and 17.5%, respectively, and were not statistically significantly related to any demographic characteristics, obstetric history, or chorionicity. Sensitivity to predict spontaneous preterm delivery was 100%, 80%, 47%, and 35% for 28, 30, 32, and 34 weeks, respectively, for cervical length up to 25 mm. The corresponding sensitivity values for cervical lengths up to 15 mm were 50%, 40%, 24%, and 11%. The rate of spontaneous delivery at or before 32 weeks increased exponentially with decreasing cervical length at 23 weeks, from 2.9% at or greater than 46 mm, to 4.3% at 36-45 mm, 6.7% at 26-35 mm, 31% at 16-25 mm, and 66% at 15 mm or less. CONCLUSION: Measurement of cervical length in twin pregnancies predicted risk of spontaneous early preterm delivery. PMID- 10472877 TI - Randomized, double-masked comparison of oxytocin dosage in induction and augmentation of labor. AB - OBJECTIVE: To test the hypothesis that high-dose oxytocin, when used in a masked fashion, would result in shorter labors and less need for cesarean delivery. METHODS: We conducted randomized, double-masked trials of high-dose compared with low-dose oxytocin for augmentation and induction of labor. Patients were randomly assigned to receive oxytocin by either a low-dose protocol (1.5 mU/minute initially, increased by 1.5 mU/minute every 30 minutes) or a high-dose protocol (4.5 mU/minute initially, increased by 4.5 mU/minute every 30 minutes). Oxytocin solutions were prepared by a central pharmacy and infusion volumes (mL/hour) were identical, thus ensuring double masking. RESULTS: A total of 1307 patients were randomized (induction, 816; augmentation, 491). In the group receiving oxytocin for induction, high-dose oxytocin was associated with a significant shortening of labor (oxytocin to complete dilatation: 9.7+/-0.3 compared with 7.8+/-0.2 hours, P<.001; oxytocin to delivery: 10.5+/-0.3 compared with 8.5+/-0.3 hours, P<.001). The cesarean delivery rate with low-dose oxytocin was 15.0%, compared with 11.3% with high-dose oxytocin (P = .17). For nulliparous women undergoing induction, cesarean delivery rates were as follows: Total 17.3% (low dose) compared with 11.7% (high dose), P = .15; cephalopelvic disproportion 11.9% (low dose) compared with 5.9% (high dose), P = .06. When used for augmentation, high-dose oxytocin again was associated with a significant shortening of labor without a significant difference in cesarean birth rates. No differences in neonatal outcomes were noted between the groups for either augmentation or induction. CONCLUSION: When used in a double-masked fashion, high-dose oxytocin is associated with significantly shorter labors without any demonstrable adverse fetal or neonatal effects. PMID- 10472878 TI - Nucleated red blood cells in maternal blood during pregnancy. AB - OBJECTIVE: To assess the frequencies of nucleated red blood cells (RBCs) in maternal blood during different stages of gestation and postpartum. METHODS: Peripheral venous blood samples were collected longitudinally from 38 pregnant women from the first trimester to 3 months postpartum. Nucleated RBCs were isolated by using a triple-density gradient with Histopaque (Sigma Diagnostics, St. Louis, MO) and identified by Kleihauser-Betke acid stain. RESULTS: The number of nucleated RBCs steadily increased from 3.9 (standard error 0.6) per 10(7) nucleated cells in early gestation (6-10 weeks) to 112.0 (standard error 7.5) per 10(7) nucleated cells near term and decreased rapidly after delivery. The number of nucleated RBCs was not related to the gender of the fetus or the ABO blood type compatibility between the mother and fetus. CONCLUSION: The number of nucleated RBCs in the maternal blood increase progressively throughout pregnancy, with some variation from subject to subject. PMID- 10472879 TI - Blunt-edged, notched scalpel for cesarean incision. AB - BACKGROUND: Fetal penetrating injuries during cesarean delivery are rare but possible complications. We describe our findings from using a newly developed blunt-edged, notched scalpel to incise the uterine wall during cesarean delivery. INSTRUMENT: The scalpel was designed with a blunt edge and notches. The blunt edge did not penetrate the uterine wall, and thus fetal injury was prevented. The notched blade split the uterine muscle fibers when the entry incision was made. EXPERIENCE: We used this tool in 41 women with intact membranes who had uterine wall incisions during cesarean deliveries. There were no fetal injuries or major complications. CONCLUSION: Making uterine incisions with this new blunt-edged, notched scalpel is feasible, easy, and safe. PMID- 10472880 TI - Bundling a pregnancy test with the Yuzpe regimen of emergency contraception. AB - The recent United States Food and Drug Administration approval of a commercial kit containing the Yuzpe regimen for emergency contraception is a welcome event. Unlike emergency contraceptive pills sold in other countries, however, the United States product has a pregnancy test bundled with the pills. The test could identify existing pregnancies and avoid unnecessary use of the pills, although any protection against lawsuits alleging injury to an embryo is speculative. Conversely, no major medical organization recommends routine pregnancy testing before using emergency contraceptive pills. The test might stigmatize the Yuzpe regimen as being dangerous to an embryo. Difficulty in understanding the pregnancy test instructions could, paradoxically, deter some women from using the pills after having bought them. The bulky size of the pregnancy test reagent stick makes the package indiscreet, and the test adds unnecessary cost to emergency contraception. The greatest usefulness of the test could be to confirm or exclude a pregnancy several weeks after taking the pills, rather than before. If bundling an unnecessary test with emergency contraception is the only way to bring this useful product to the United States market, then the public health benefits could outweigh the disadvantages. However, this approach sets a worrisome precedent and further isolates the United States from the international medical community. PMID- 10472881 TI - A simple, inexpensive device for teaching the loop electrosurgical excision procedure. AB - OBJECTIVE: To develop an inexpensive and practical inanimate model to teach the loop electrosurgical excision procedure in a resident clinic. TECHNIQUE: A vaginal speculum is inserted into one end of a cardboard tube. At the opposite end, a piece of knockwurst sausage is inserted into the cardboard tube. A grounding pad is attached to the distal end of the sausage. The teaching session proceeds using a colposcope or direct visualization. After the training session, the excised and uncut portions of sausage can be examined. EXPERIENCE: Faculty found this inanimate model an excellent way to teach residents how to use the loop electrode, and the resident staff appreciated its merits as a useful prelude to treating cervical disease in women. The house staff adapted to the confines of the vagina and transferred skills acquired from working with the cardboard tube, where instruction and constructive criticism can be given without the presence of apprehensive patients. CONCLUSION: A simple, inexpensive, inanimate model has been developed to teach the loop electrosurgical excision procedure. The materials required to construct it are available to any colposcopy clinic. PMID- 10472882 TI - Practice management in obstetrics and gynecology residency curriculum. AB - OBJECTIVE: To establish the current level of instruction in practice management in obstetrics and gynecology residency programs; review recommendations of medical school practice management executives and current and former residents on design of practice management instruction programs; and develop recommendations for future management instruction by residency programs in obstetrics and gynecology. METHODS: A questionnaire was sent to 101 practice plan executives of obstetrics and gynecology departments at medical schools in the United States and Canada. A modified version was sent to 44 current obstetrics and gynecology residents and 72 former residents from the University of Tennessee, Memphis. RESULTS: The response rates were 71% (practice executives), 93% (current residents), and 81% (former residents). There were no formal management programs at 87% of responding institutions, although most respondents (62%) thought there should be mandatory participation in management programs, probably given by organizations outside the university. Potential subjects that received high ratings were current procedural terminology and diagnosis coding, managed care, billing procedures, contractual agreements between medical doctors, patient record management, and practice economics. CONCLUSION: Residency programs should establish formal practice management instruction programs and make participation mandatory. Funding should come from the medical school and university. Instructional help should come from extradepartmental organizations and individuals. PMID- 10472883 TI - Fetal amino acid and enzyme levels with maternal smoking. PMID- 10472884 TI - Failure to obtain follow-up testing for gestational diabetes in a rural population. PMID- 10472885 TI - Amniotic fluid lamellar body count: cost-effective screening for fetal lung maturity. PMID- 10472886 TI - Associated factors in 1611 cases of brachial plexus injury. PMID- 10472887 TI - Immunisation of preterm babies. PMID- 10472888 TI - Protecting the health of medical students on electives. PMID- 10472889 TI - Persistence of iodine deficiency 25 years after initial correction efforts in the Khumbu region of Nepal. AB - AIMS: To assess the current status of, and understanding about iodine deficiency disorders among Sherpa residents of the Khumbu region of Nepal, 25 years after the introduction of iodised oil injections. METHODS: Several groups of Khumbu Sherpas were studied and goitre rate, urinary iodine level and cretinism prevalence were measured as indicators of iodine deficiency. Subjects were also questioned in detail about their food consumption, with particular reference to salt use, and about their understanding of the causes and treatment of iodine deficiency disorders. RESULTS: The prevalences of goitre, deaf-mutism and cretinism were 21%, 1.3% and 0.5% respectively (compared to 92%, 4.7% and 5.9% in 1966). No cretins had been born since 1966. The median urine iodine concentration was 35 microg/L. Most people preferred uniodised Tibetan rock salt, although 44% regularly consumed iodised salt. All granulated salt tested from the local market contained adequate amounts of iodine. Only 11% of those surveyed knew that goitre was caused by iodine deficiency CONCLUSIONS: Although prevalences of iodine deficiency disorders are much less than 30 years ago, iodine deficiency continues to be a major problem in Khumbu and demands a clear control strategy, combining ongoing iodine supplementation and education. Iodised salt is usually the best approach to control of iodine deficiency disorders for most regions of the world but the Khumbu experience shows that local cultural and commercial factors can severely limit its impact. To be successful, control programme for iodine deficiency disorders also needs assessment of the salt trade, monitoring, education and occasional targeted interventions with iodised oil or other supplements. PMID- 10472890 TI - Mortality after hip fracture: regional variations in New Zealand. AB - AIM: To determine the 35-day and one-year mortality rates following a hospital admission for hip fracture, among individuals aged 60 years or older in New Zealand. METHODS: New Zealand Health Information Service mortality data for the years 1988 to 1992 were examined to determine the case fatality rate among individuals aged 60 years or older admitted to hospital for fractures of the neck of femur (ICD-9 N-code 820). Case fatality rates assessed at 35 days and one year after admission to hospital were examined by age, gender, year of admission, place of residence, area health board region and cause of death. RESULTS: Between 1988 and 1992, the case fatality rate was 8% within 35 days of admission to hospital and 24% within one year of admission. Case fatality rates were found to be twice as high in men compared to women and four to five times higher in individuals aged 85 years and older, compared to people aged between 60 and 64 years. The only regional difference in hip fracture mortality was found in the Canterbury area health board region, which had a 30% higher rate of hip fracture mortality compared to all regions combined. The two main cited underlying causes of death after hip fracture were accidental falls (ICD E880-E888) and ischaemic heart disease (ICD 410-414). CONCLUSION: Over three-quarters of individuals aged 60 years or older who are hospitalised with a hip fracture in New Zealand survive for at least one year after admission. However, significant variations in mortality exist with age and gender. These data highlight the importance of preventive strategies for hip fracture in older people and the need to identify ways of improving post-admission care. PMID- 10472891 TI - Outcome of managing impotence in clinical practice. AB - AIMS: Outside of controlled clinical trials, the outcome of treatment for unselected men with impotence is uncertain. This study aims to describe the clinical course of consecutive, unselected men referred to a specialist endocrinology private practice with a primary diagnosis of impotence. METHODS: Consecutive men referred with a primary diagnosis of impotence between June 1995 and December 1997 were studied. After initial evaluation and appropriate investigation, treatment with testosterone in hypogonadal men and instruction in the use of a vacuum device and intracavernosal alprostadil (Caverject) in all men was offered. All men were followed up by telephone and/or questionnaire about erection outcome three to twelve months later. RESULTS: Nineteen diabetic men, aged 53.1+/-8.2 years and forty non-diabetic men, aged 54.8+/-11.6 years were seen. Follow-up information beyond three months was complete in fifty-three (90%). Eighteen eugonadal men chose no further therapy and four of these men had spontaneous return of erections. Eight men were hypogonadal and potency returned in two of six men treated with replacement testosterone. Nine men used the vacuum device, which was effective in three of them. Forty-one men had a trial of Caverject injection, which was effective in twenty-eight. Only twelve of these men used Caverject for longer than six months. CONCLUSIONS: Return of erections with therapy beyond three months in unselected men with impotence is successful in only about one-third. Unexpected hypogonadism is relatively common in impotent men, but testosterone replacement therapy has a low rate of improving erections. New therapies for impotence need careful follow-up studies to assess their effectiveness in clinical practice. PMID- 10472892 TI - Did Wellington have too many acute psychiatric beds in 1996? AB - AIMS: (1) To establish whether, as believed by local clinicians, there is a shortage of acute psychiatric beds available in Wellington. (2) To compare the current situation with that of seven years ago, before the beginning of the reduction in bed numbers, as a part of deinstitutionalisation and care in the community. (3) To study the effect on occupancy of changing availability. METHOD: Data were obtained retrospectively for the calendar years 1996 and 1989. These were examined using an analysis of variance and the results of the two years were compared. RESULTS: There were 214 days in 1996 when there were no acute psychiatric beds available. The mean daily occupancy was 100%. Comparing like populations, there was a highly significant increase in the mean occupancy as the availability decreased from 1989 to 1996. Conversely, there was a decrease in occupancy as availability temporarily increased during 1996. CONCLUSION: The number of acute psychiatric beds available in Wellington in 1996 was insufficient to meet the needs of the population, given the alternative resources available at that time. This is of particular concern in light of proposals to reduce the number of beds further. PMID- 10472893 TI - D-lactic acidosis in short bowel syndrome. AB - Short bowel syndrome can present many complex management issues and may be complicated by various metabolic problems. D-lactic acidosis in the setting of short bowel syndrome has been described only rarely in children. A further case is presented with a review of typical clinical manifestations of D-lactic acidosis and reported management options. Early recognition and appropriate management is essential to avoid morbidity secondary to this complication of short bowel syndrome. Probiotic therapies may have an increasing role in prevention and management of this complication. PMID- 10472894 TI - The Stent report, one year on. Christchurch Hospital's Medical Specialists Association. PMID- 10472895 TI - Hypertension guidelines for diabetic patients. PMID- 10472896 TI - Youth suicide. PMID- 10472897 TI - Psychological factors in illness. PMID- 10472898 TI - Ciguatera fish poisoning. PMID- 10472899 TI - Failure to diagnose cervical cancer. PMID- 10472900 TI - The history of the EMST course. Early Management of Severe Trauma. PMID- 10472901 TI - What makes nurses important to EMST and EMST important to nurses? Early Management of Severe Trauma. PMID- 10472902 TI - EMST in Papua New Guinea: more than just a course. Early Management of Severe Trauma. AB - BACKGROUND: Trauma in Papua New Guinea (PNG) is a major reason for acute hospital admission. The Early Management of Severe Trauma Course (EMST) of the Royal Australasian College of Surgeons was introduced to PNG in 1993 to help in trauma education. METHODS: Four EMST courses have been held in Port Moresby since 1993 with the support of the Department of Surgery of the University of PNG. RESULTS: Sixty-two doctors have attended a course and three PNG doctors have been trained as course instructors. Trauma workshops were held in other provincial hospitals extending the course principles to nurses and health professionals in these areas. CONCLUSION: The introduction of EMST to PNG has proven to be a worthwhile endeavour and the program is now firmly established in PNG. PMID- 10472903 TI - EMST scholarship report: experience in Washington DC and Boston. Early Management of Severe Trauma. PMID- 10472904 TI - EMST scholarship report: experience in South Carolina compared with Echuca. Early Management of Severe Trauma. PMID- 10472905 TI - The John Graham Extrication Prize: its history and importance. AB - Management of Severe Trauma Coursersef Severe Trauma (EMST) course is unique in providing instructive exposure to techniques used in the extrication of severely injured victims from difficult locations using a role play method. Dr John C. Graham FRACS, vascular surgeon in Lismore, was responsible for the initiation of this concept in 1989. Since this component of the trauma course is not used in the Advanced Trauma Life Support (ATLS) course in the USA, from which the EMST course was derived, the purpose of the present study is to determine the opinions of all EMST course directors as to the value of including an extrication scenario station in the EMST course in its current form in Australia and New Zealand. METHODS: The extrication station as devised by John Graham consists of four 40 min scenarios offered to all 16 participants of an EMST course in groups of four. The participants are required to extricate a moulaged victim from a difficult location within the time available in the most skilful and expeditious manner. A prize is offered for the group performing the task best. A survey of the 45 EMST course directors was undertaken by questionnaire. Replies were anonymous. RESULTS: There was a good response rate of 87%. Ninety-two per cent of responding directors routinely used an extrication station. Two directors used the station occasionally and only one director never used it. The most frequently cited reason for use was that the participants come to appreciate commonly encountered problems in extrication, and how EMST principles are applied in the field. Second, the station allows the use of teamwork. Third, it adds entertainment to the course overall. There was a prize awarded by every course director for the winning group of participants. Ninety per cent of these prizes were named the 'John Graham Extrication Prize'. CONCLUSIONS: The extrication station is considered constructive by the great majority of EMST directors. Both historical and competitive aspects are important since most directors still award a named prize for this station. The extrication station promotes a feeling of cohesion among Faculty and participants on an EMST course. The station is a memorable event for many participants and should be retained as a uniquely Australasian contribution to the course. PMID- 10472906 TI - ATLS: are we training the wrong people? AB - BACKGROUND: Despite the evolution, expansion and popularity of emergency medicine as a medical specialty in the United Kingdom (UK), emergency departments are still primarily staffed by senior house officers (second and third year graduates), particularly at weekends and at night. METHODS: A telephone and postal survey of Advanced Trauma Life Support (ATLS) training completed by this cohort of doctors indicates that UK courses are failing to train adequate numbers. RESULTS: It can thus be argued that ATLS is not being disseminated to the relevant grass-roots of medical care, where it is likely to be of the most benefit. CONCLUSIONS: The present study discusses possible reasons for this and offers constructive solutions to the problem. Although the matters discussed in this study refer to UK medical practice, they may be of relevance and interest to Australasian practitioners. Is EMST in Australasia training the appropriate group of doctors? PMID- 10472907 TI - Port Arthur disaster. PMID- 10472908 TI - Black Hawk helicopter mid-air collision: Townsville General Hospital perspective. PMID- 10472909 TI - The crash of flight 703: a hospital's response. PMID- 10472910 TI - An Australasian perspective of chest trauma. PMID- 10472911 TI - A review of complex trauma to the elbow. AB - This paper presents a review of common complex elbow traumatic disorders. It presents an algorithm to aid in the management of these cases and discusses indications for radial head replacement and dynamic external fixateurs. PMID- 10472912 TI - Ocular trauma. PMID- 10472913 TI - Trauma in the elderly: a new epidemic? AB - Australia's population is ageing. Statistics predict that by the year 2021, 17.8% of our population will be over the age of 65 years. These statistics are reflected in most Western nations. The elderly represent an increasing proportion of emergency department patients. They are generally more acute on presentation, they get admitted more frequently, and account for an increasing proportion of admissions to intensive care units. The elderly trauma patient will increasingly consume more resources than patients from any other age group. There are significant differences in physiology and response to trauma between the elderly patient and the young. Airway compromises, a decrease in lung compliance, a change in cardiovascular haemostasis and the prevalence of pre-existing disease all contribute to increase morbidity and mortality in a patient with an already limited physiological reserve. Many studies demonstrate an increased mortality in hospitalized elderly trauma patients. For those who die from traumatic insult, death is mainly due to multiorgan system failure. The long-term functional outcome of the elderly trauma patient has been investigated by a number of authors. One study showed that only 8% of survivors returned to independent living 1 year post-polytrauma. Another more encouraging study showed that 89% of survivors of blunt multiple trauma after 38 months were independent and living at home. Ethical issues have been raised regarding the appropriateness of care for the severely injured elderly patient given today's ever-shrinking hospital finances and bed shortages. Today's environment is very challenging as frequently, the clinician is forced to make an economic decision on who will take the last critical care bed, which is a far from optimal situation. PMID- 10472914 TI - Is the 'tunnel of death' a suitable modality for investigating the severely traumatized child? AB - BACKGROUND: The child who has suffered severe physical trauma and is physiologically unstable can often present a diagnostic dilemma to the clinician in the emergency room. METHODS: In an attempt to clarify the situation there is often a decision made to perform computed tomographic scanning in such children. RESULTS: This is an extremely dangerous investigation in this situation given that the child has to have a GA as well as being unstable from the injuries. CONCLUSION: Ultrasound is easier, less expensive, can be performed in the emergency room and can be performed by relatively inexperienced operators with good diagnostic accuracy. PMID- 10472915 TI - Troublesome trauma topics: paediatric problems. PMID- 10472916 TI - The application of ultrasound in the assessment of the trauma patient. PMID- 10472917 TI - Ultrasound assessment of the trauma patient. PMID- 10472918 TI - The doctor's emergency kit. PMID- 10472919 TI - Australian disaster triage: a colour maze in the Tower of Babel. AB - BACKGROUND: The objective of this study was to review the systems of disaster triage used by Australian State and Territory ambulance services and compare their triage taxonomy, methodology and documentation with the Australian Council on Health Care Standard's (ACHCS) National Triage Scale, which is used in all Australian hospital emergency departments. METHODS: A postal survey of the State and Territory ambulance services during October 1996 was conducted. Details of the mass casualty incident (MCI) triage systems were then compared with the ACHCS National Triage Scale. Colours specified or used on a triage tag were checked for compliance with Standards Australia AS-2700 1996 Colour Standards for General Purposes. Participants consisted of those State and Territory ambulance services which would be the initial emergency medical service responders in the event of an MCI in an Australian capital city, and the ACHCS. The main outcome measure was the homology between the respective triage taxonomies, methodologies and documentation systems. RESULTS: All eight State and Territory ambulance services used a numerical and colour coded system to indicate triage priority during an MCI. There were five different triage tag designs for triage documentation, six different triage taxonomies and five different triage methodologies with minimal homology between the different triage systems and the National Triage Scale used in hospitals. Only two ambulance triage systems specifically triaged emotional disturbance. Several triage tags and their patient attachments were made from perishable materials and are thus likely to fail under field conditions. CONCLUSION: The multiplicity of triage systems used within Australia will result in avoidable confusion, thus hindering the medical response to an MCI, especially for incidents near State or Territory borders. There is little evidence to support the continued use of triage tags. Australia needs to develop a uniform system of patient triage as a national priority. PMID- 10472920 TI - An Australian mass casualty incident triage system for the future based upon triage mistakes of the past: the Homebush Triage Standard. AB - BACKGROUND: The purpose of this study was to create a system of mass casualty incident triage that provides a common language platform for both ambulance and hospital personnel based upon standard daily operating procedures for both groups. METHODS: An annotated literature review was conducted. RESULTS: The Homebush Triage Standard taxonomy consists of five categories: immediate, urgent, not urgent, dying and dead, which are given the phonetic alphabet designations of Alpha, Bravo, Charlie, Delta and Echo, respectively, to facilitate radio voice communications. Colours are assigned to each category: red (Homebush Red), yellow (Homebush Gold), green (Homebush Green), white and black comply with Standards Australia AS-2700 1996 Colour Standard for General Purposes, to ensure triage materials have consistent production standards. Numbers are reserved to either quantify or stratify individuals within a particular triage category. The Homebush Triage Standard methodology is based on START (Simple Triage Assessment and Rapid Transport) and SAVE (Secondary Assessment of Victim Endpoint) and documents triage priority using geographic triage with triage flags, instead of triage tags. CONCLUSION: The Homebush Triage Standard provides a basis for both seamless patient reassessment at all points along the casualty evacuation chain and the audit of the medical response to mass casualty incidents. It allows hospital and ambulance staff to keep working using familiar routines and removes fundamental barriers to good communication during a time of crisis. PMID- 10472921 TI - The management of blunt liver injuries. AB - Although the general surgeon who takes emergency call may be confronted with a patient who has sustained a blunt liver injury, the decrease in road trauma and work-place accidents has meant that this will be an infrequent occurrence. Minimal exposure will, of necessity, extrapolate to difficulty in coping with a catastrophic event which comes unheralded, at an inconvenient time and usually when there is less than ideal support. During the past 15 years, there has been an evolution in the treatment of liver injuries which is exemplified by a non operative approach in selected patients and more conservative procedures in those who require operative intervention. At present, 'damage control' is in vogue; do the least possible to control life-threatening injuries and come back another day. This is a cogent and admirable philosophy, provided that the pendulum does not swing too far and that a planned course of action is in place. This paper reviews the present status of managing blunt liver injuries, with an emphasis on the general surgeon who has little cause to be involved with surgery of the liver. PMID- 10472922 TI - Complete removal of nonpalpable breast malignancies with a stereotactic percutaneous vacuum-assisted biopsy instrument. AB - BACKGROUND: Several devices have been developed for sampling nonpalpable mammographic breast lesions. Complete removal of malignancies with a stereotactic percutaneous directional vacuum-assisted biopsy instrument has been reported. STUDY DESIGN: We reviewed our experience with the percutaneous vacuum-assisted biopsy instrument to identify instances of complete excision of cancers: no residual carcinoma found at surgical excision for malignancies diagnosed by the percutaneous vacuum-assisted biopsy instrument. The radiologic and pathologic characteristics of malignancies completely removed by the percutaneous vacuum assisted biopsy instrument were compared with those of malignancies with residual carcinoma found at surgical excision. RESULTS: Fifty-two malignancies were diagnosed by the percutaneous vacuum-assisted biopsy instrument: 16 infiltrating ductal carcinomas, 5 infiltrating lobular carcinomas, and 31 ductal carcinomas in situ. No residual carcinoma was found at surgical excision in 9 (17%) of the 52 malignancies. Patients with complete removal of the malignant lesion were younger than patients with incomplete removal (52 versus 58 years; p = 0.069). Completely removed malignancies were smaller on mammography (4 versus 17 mm; p = 0.213), and more specimens were removed (19 versus 15; p = 0.074). All nine completely removed malignancies presented with calcifications without a mass (p = 0.112), and all nine were ductal carcinoma in situ (p = 0.019). CONCLUSIONS: Complete removal of nonpalpable breast malignancies is possible with the stereotactic percutaneous directional vacuum-assisted biopsy device. Complete removal is more likely with removal of a large number of specimens from small areas of mammographic calcifications due to ductal carcinoma in situ. PMID- 10472923 TI - Intraoperative ultrasound localization to guide surgical excision of nonpalpable breast carcinoma. AB - BACKGROUND: This report describes a technique of intraoperative tumor localization by ultrasound without the use of a needle or wire to guide the excision of nonpalpable breast cancers. The results of our experience with pathologic margin status are reviewed. STUDY DESIGN: From 1994 to 1998, 65 breast cancers in 62 patients with biopsy-proved nonpalpable breast cancer were excised using intraoperative ultrasound localization. The pathologic status of the margins from the initial surgical excision specimen and any further excisions, either at the first operation or later procedures, was recorded. The distance from the tumor to the closest margin of excision was also determined. RESULTS: The overall success in achieving pathologically negative excision margins at first operation was 97% (63 of 65 cancers). Three patients underwent a second operative procedure, two for positive margins and one for a margin less than 1 mm (second operation = 4.8% of patients). After completion of the first operative procedure, the mean distance to the closest margin of excision was 0.8 cm. CONCLUSIONS: Intraoperative ultrasound localization for excision of nonpalpable breast cancers is feasible and gives results, in terms of pathologic margins, that are comparable with those achieved by standard needle-wire-guided excisions. PMID- 10472924 TI - Lymphatic mapping and sentinel lymphadenectomy after wide local excision of primary melanoma. AB - BACKGROUND: Lymphatic mapping and sentinel lymphadenectomy (LM/SL) are generally avoided in patients who have already undergone wide local excision (WLE) of a primary melanoma, because of concern that disruption of the cutaneous lymphatics might alter lymphatic flow to the sentinel node. We reviewed carefully chosen patients who had undergone LM/SL after WLE to identify circumstances that might make this approach otherwise safe and clinically accurate. STUDY DESIGN: From our melanoma database of 8,300 patients, of whom 1,015 had undergone LM/SL, we retrospectively identified 47 patients who had previously undergone WLE. Patient and tumor characteristics were collected and compared with followup data from clinic files. RESULTS: Median WLE surgical margins before LM/SL were 2.0 cm and most patients had extremity lesions. Eleven of the 47 patients (23%) had tumor involved sentinel nodes, and 8 of these patients (73%) had a solitary nodal metastasis. With a median followup period of 36 months, 3 sentinel node-negative patients developed nodal recurrences. Two of these patients had positive sentinel nodes on pathology re-review and were not considered failures of the lymphatic mapping surgical procedure. The third patient developed in-transit metastases and delayed nodal recurrence. An additional patient, who had a primary tumor on the trunk, developed a nodal recurrence in the basin opposite that identified by lymphoscintigraphy. The overall error rate of the technique was 4 in 36 (11%). This included 2 pathology misdiagnoses (5.6%), 1 nodal recurrence associated with in-transit regional metastases (2.8%), and 1 lymphatic mapping error (2.8%). CONCLUSIONS: LM/SL can be cautiously performed in patients who have undergone previous WLE if the primary resection margin was no greater than 2.0 cm and the primary was not in a region of ambiguous drainage. Lymphatic mapping may be inaccurate when melanomas have been resected with large margins, especially if the wound was closed with rotation flaps, and when melanomas are on the head and neck or trunk regions. PMID- 10472925 TI - Intraoperative decision making in follicular lesions of the thyroid: is tumor size important? AB - BACKGROUND: Intraoperative decision making in treating follicular lesions of the thyroid remains controversial because there are no reliable preoperative or intraoperative factors predictive of malignancy. This study was undertaken to determine whether lesion size is a reliable factor that can be used to predict a final pathologic diagnosis of follicular carcinoma. STUDY DESIGN: This was a retrospective, case-matched control study. One hundred consecutive patients with follicular carcinoma were matched by gender, age, and date of operation with 100 patients with follicular adenomas. Seventy-nine matched pairs had pure follicular lesions and 21 matched pairs had oxyphilic variants of follicular lesions. After confirming adequate matching, lesion size was compared between groups. RESULTS: Regardless of whether all follicular lesions were analyzed or whether only pure follicular or oxyphilic variant lesions were compared, there was no significant difference in lesion size between the carcinoma and adenoma groups. The mean size of all follicular carcinomas was 31.5 +/- 1.7 mm and the mean size of all follicular adenomas was 30.8 +/- 1.5 mm (p = NS). When the proportions of the carcinoma and adenoma groups were indexed by five different size intervals and compared, there was again no significant difference in any category. CONCLUSIONS: On the basis of this case-matched control study, the size of a follicular lesion cannot be used to predict a final diagnosis of follicular carcinoma and is of no value when making intraoperative decisions about the extent of thyroid resection. PMID- 10472926 TI - A new approach to the palliation of advanced proximal gastric cancer. AB - BACKGROUND: The operative mortality and morbidity rates of palliative total gastrectomy can be high, and survival is not extended. The quality of a foreshortened life is often marred by distressing postprandial symptoms and relentless weight loss. These problems can be attributed to the conventional manner of reconstruction after total gastrectomy, with small-bowel gastric reservoirs restricting the amount of caloric intake. Large-bowel gastric reservoirs have greater capacity and empty well if positioned upright within the chest for proper emptying. The scope of a combined palliative total gastrectomy, esophagectomy, and colon pull-through must be kept within the limits of patient tolerance. STUDY DESIGN: A palliative total gastrectomy was performed in 70 patients with incurably advanced cancers of the proximal and middle third of the stomach (TNM stages: II, 4%; III, 26%; and IV, 70%) using the large-bowel as a gastric substitute. The trauma of reconstruction by colon pull-through was lowered by avoiding thoracotomy and by positioning the colon within the lumen of the deepithelialized esophagus. Proximal cervical esophagocolostomy, distal duodenocolostomy, and colocolostomy reestablished gastrointestinal continuity. Follow-up focused on subjective gastrointestinal symptoms and nutritional maintenance. RESULTS: The operative mortality rate was 10%, postoperative complications were not inordinately high, and autopsy findings showed no defects in the technique of reconstruction. The normal esophageal mucosa was readily cored out through the neck and the abdomen, and the remaining esophageal muscular tunnel accommodated the pulled-through segment of colon. Quantitative assessment of postoperative quality of life proved impractical, but none of 58 longterm survivors (mean of 17.1 months for combined TNM stages II, III, and IV) suffered from disabling symptoms or pronounced weight loss. The quality of life, but not its length, appeared distinctly improved. CONCLUSIONS: The frequently encountered problems of abdominal distress and weight loss after palliative total gastrectomy can be averted by safely positioning a colonic gastric substitute within the lumen of the normal esophagus from which the mucosal lining has been extracted. PMID- 10472927 TI - Acquired abnormalities of the biliary tract from chronic gallstone disease. AB - BACKGROUND: Acquired abnormalities of the biliary tract from chronic gallstone disease are rare. The aim of this study was to examine the frequency with which these abnormalities occur and to assess the probability of encountering such an abnormality at laparoscopic cholecystectomy. STUDY DESIGN: We conducted a prospective study of all patients undergoing elective and emergency cholecystectomy under the care of one surgeon between January 1991 and December 1997. RESULTS: Biliary tract abnormalities from chronic gallstone disease were encountered in 10 (2%) of 486 patients undergoing cholecystectomy. Four were observed in patients undergoing elective laparoscopy cholecystectomy, and the remainder were observed at open cholecystectomy. Five had a cholecystocholedochal fistula (Mirizzi Syndrome Type II), and one had a stone impacted at the cystic duct-bile duct junction (Mirizzi Syndrome Type I). Two had cholecystoduodenal fistulas and two had an absent cystic duct with a normal bile duct. Both instances of an absent cystic duct were encountered at laparoscopic cholecystectomy; in one the bile duct was mistaken for the cystic duct and a 2-cm segment was excised at operation, and in the other the abnormality was recognized and confirmed by cholangiography. CONCLUSIONS: This study demonstrates a similar incidence of acquired abnormalities of the biliary tract from chronic gallstone disease to that already reported. But acquired absence of the cystic duct may occur more frequently than previously suspected. Patients with this condition are at high risk for bile duct injury during laparoscopic cholecystectomy. Clinical awareness of this problem with strict adherence to the principles taught at open cholecystectomy may prevent or reduce the severity of bile duct injury in these patients. PMID- 10472928 TI - The prevalence and natural history of gallstones in spinal cord injured patients. AB - BACKGROUND: Individuals with a spinal cord injury are at increased risk for the development of gallstones. Because these patients cannot reliably manifest classic symptoms of biliary colic, they may be more likely to present with advanced biliary complications than patients with intact abdominal innervation. The natural history of gallstones in spinal cord injured patients has not been described. STUDY DESIGN: All spinal cord injured patients seen at the Seattle Veterans Affairs Medical Center from January 1, 1993, to December 31, 1997 were included in the study. For each patient, the presence or absence of gallstones had been determined previously through screening abdominal ultrasonographic evaluations. Pertinent demographic information was obtained from medical records and patient interviews. Patients with gallstones were followed until death, cholecystectomy, or the conclusion of the study, and the annual incidence of biliary complications and patients requiring a cholecystectomy were determined. The prevalence of gallstones was established by studying the subset of patients seen at the Seattle Spinal Cord Injury Unit from January 1, 1995 to December 31, 1997. RESULTS: Among the spinal cord injured patients, 31% either had gallstones or had undergone a cholecystectomy at some point after their injury. Increasing age, female gender, and greater severity of injury were risk factors for the formation of gallstones. Over the first 5 years after the diagnosis of gallstones, the annual incidence of cholecystectomy or biliary complications was 6.3% and 2.2%, respectively. CONCLUSIONS: Spinal cord injured patients are at increased risk for the development of gallstones. Patients with gallstones are at an increased risk for the development of biliary complications compared with neurologically intact patients, but the magnitude of this risk does not warrant prophylactic cholecystectomy. PMID- 10472929 TI - Resection of hepatocellular carcinoma in cirrhotic patients: longterm results of a prospective study. AB - BACKGROUND: Surgical resection of hepatocellular carcinoma in cirrhotic patients remains controversial because of a high reported recurrence rate. To assess the longterm results of resection, 37 patients included in a prospective study were followed for more than 5 years, with special interest in early detection of recurrence. STUDY DESIGN: Resection was performed from 1986 to 1991 with the goal of sparing the functional liver parenchyma. The mean tumor diameter was 5.3 +/- 2.6 cm (range 2 to 11 cm). Nineteen patients had tumors smaller than 5 cm. No additional perioperative therapy was performed. RESULTS: Evidence of intrahepatic recurrence was demonstrated in 26 of the 33 patients surviving the operation. Eight recurrences (31%) were diagnosed from the third to the fifth postoperative years. The recurrence-free survival rates at 1, 2, 3, 4, and 5 years were 68%, 40%, 26%, 13%, and 9%, respectively. Only 2 patients (7%) were alive and free of recurrence at 5 years. Some long survivals were observed after treatment of recurrence. The overall survival rates at 3 and 5 years were 35% and 24%, respectively. Tumor cell differentiation was the only significant prognostic factor for both recurrence and survival. Multifocal tumors were associated with a higher recurrence rate. Patients with good liver function had longer survivals that reached 38% in those with small solitary tumors. Study of the other dinicopathologic factors failed to demonstrate any prognostic value. CONCLUSIONS: Only a few patients are alive and free of recurrence 5 years after resection. Some long survival can be observed after treatment. Assessment of prognostic factors remains difficult, but the best results of resection are obtained in patients with small solitary hepatocellular carcinoma function. PMID- 10472930 TI - Hepatic resection for metastatic colorectal adenocarcinoma: a proposal of a prognostic scoring system. AB - BACKGROUND: Hepatic resection for metastatic colorectal cancer provides excellent longterm results in a substantial proportion of patients. Although various prognostic risk factors have been identified, there has been no dependable staging or prognostic scoring system for metastatic hepatic tumors. STUDY DESIGN: Various clinical and pathologic risk factors were examined in 305 consecutive patients who underwent primary hepatic resections for metastatic colorectal cancer. Survival rates were estimated by the Cox proportional hazards model using the equation: S(t) = [So(t)]exp(R-Ro), where So(t) is the survival rate of patients with none of the identified risk factors and Ro = 0. RESULTS: Preliminary multivariate analysis revealed that independently significant negative prognosticators were: (1) positive surgical margins, (2) extrahepatic tumor involvement including the lymph node(s), (3) tumor number of three or more, (4) bilobar tumors, and (5) time from treatment of the primary tumor to hepatic recurrence of 30 months or less. Because the survival rates of the 62 patients with positive margins or extrahepatic tumor were uniformly very poor, multivariate analysis was repeated in the remaining 243 patients who did not have these lethal risk factors. The reanalysis revealed that independently significant poor prognosticators were: (1) tumor number of three or more, (2) tumor size greater than 8 cm, (3) time to hepatic recurrence of 30 months or less, and (4) bilobar tumors. Risk scores (R) for tumor recurrence of the culled cohort (n = 243) were calculated by summation of coefficients from the multivariate analysis and were divided into five groups: grade 1, no risk factors (R = 0); grade 2, one risk factor (R = 0.3 to 0.7); grade 3, two risk factors (R = 0.7 to 1.1); grade 4, three risk factors (R= 1.2 to 1.6); and grade 5, four risk factors (R > 1.6). Grade 6 consisted of the 62 culled patients with positive margins or extrahepatic tumor. Kaplan-Meier and Cox proportional hazards estimated 5-year survival rates of grade 1 to 6 patients were 48.3% and 48.3%, 36.6% and 33.7%, 19.9% and 17.9%, 11.9% and 6.4%, 0% and 1.1%, and 0% and 0%, respectively (p < 0.0001). CONCLUSIONS: The proposed risk-score grading predicted the survival differences extremely well. Estimated survival as determined by the Cox proportional hazards model was similar to that determined by the Kaplan-Meier method. Verification and further improvements of the proposed system are awaited by other centers or international collaborative studies. PMID- 10472931 TI - Comparison of controlled and Glisson's pedicle transections of hepatic hilum occlusion for hepatic resection. AB - BACKGROUND: For hepatic resection, intraoperative bleeding is reduced by clamping the afferent blood flow. Selective clamping at the hepatic hilum can be accomplished using the standard controlled method or Glisson's pedicle transection method. The safety and efficacy of these two methods have not previously been compared. STUDY DESIGN: The intraoperative findings and complications were retrospectively reviewed in 90 patients who underwent major hepatectomy with selective inflow clamping between 1988 and 1997. RESULTS: Blood loss and operative time did not differ between the two groups. Bile leakage was observed in 3 of 43 patients (7.0%) in the standard controlled method group and 11 of 47 patients (23.4%) in the Glisson's pedicle transection method group (p = 0.031). In the Glisson's pedicle transection method group, bile leakage occurred more frequently in patients who underwent left lobectomy than in those who underwent right lobectomy (p = 0.023). CONCLUSIONS: There were no differences in intraoperative findings and postoperative complications analyzed between the standard controlled method and Glisson's pedicle transection method for major hepatectomy except for the rate of bile leakage. In particular, attention must be paid to the bile duct injuries when the Glisson's pedicle is dissected. PMID- 10472932 TI - Surgical complications after pancreas transplantation with portal-enteric drainage. AB - BACKGROUND: Despite recent advances, surgical complications remain an important source of morbidity after pancreas transplantation (PTX). Several previous studies have delineated the surgical complications after PTX with systemic bladder (S-B) drainage, but data are limited regarding the incidence and outcomes of surgical complications after PTX with portal-enteric (P-E) drainage. STUDY DESIGN: We retrospectively studied surgical complications after 83 vascularized PTXs with P-E drainage in 79 patients (65 simultaneous kidney-PTXs [SKPT] and 18 solitary PTXs [SPT], 8 pancreas alone and 10 pancreas after kidney transplantation). Twelve (15%) were retransplants. A surgical complication was defined as the need for repeat laparotomy within the first 3 months after PTX. RESULTS: A total of 53 surgical complications requiring repeat laparotomy occurred in 31 patients (37%). The incidence of surgical complications in SKPT and SPT was 38% and 33%, respectively. The most common indications for repeat laparotomy were: vascular thrombosis in 13% (SKPT 14% and SPT 11%), intraabdominal infection in 10% (SKPT 12% and SPT 0%), intraabdominal bleeding in 8% (SKPT 8% and SPT 11%), and duodenal allograft leak in 4% (SKPT 3% and SPT 6%). Patient survival rates at 1 and 3 years with versus without surgical complications were 84% and 80% versus 94% and 86%, respectively (p = NS). Pancreas graft survival rates at 1 and 3 years with versus without surgical complications were 48% and 44% versus 89% and 76%, respectively (p < 0.0001). The incidence of surgical complications was 45% in the first 42 P-E transplantations performed between 1990 and 1995, compared with 29% in the next 41 transplantations performed during 1996 and 1997 (p = NS). The mean number of repeat laparotomies per patient decreased from 1.2 in the former group to 0.5 in the latter group (p = NS). The incidence rates of vascular thrombosis, intraabdominal infection, and duodenal leak in the former and latter groups were 17% versus 10%, 12% versus 7%, and 2% versus 5%, respectively. CONCLUSIONS: Surgical complications after PTX are common, and their incidence and outcomes with P-E drainage are similar to those with S-B drainage. The complication rate does not vary according to the type of transplant (SKPT versus SPT). Increasing experience with P-E drainage results in a decreased incidence of surgical complications. PMID- 10472933 TI - Increased use of prophylactic vena cava filters in trauma patients failed to decrease overall incidence of pulmonary embolism. AB - BACKGROUND: Recent studies have reported that placement of vena cava filters (VCFs) early after injury may decrease the incidence of pulmonary embolism (PE) in high-risk trauma patients. STUDY DESIGN: This was a retrospective review of all trauma patients with placement of VCFs admitted to a single level-1 trauma center between 1989 and 1997. Two cohorts corresponding to years of high or low prophylactic VCF use (PVCF) were compared. RESULTS: Records were reviewed for 299 trauma patients identified as having had placement of a VCE Two hundred forty eight filters were placed before the diagnosis of PE. During years of low PVCF use, the overall PE incidence was 0.31%; during years of high PVCF use, the incidence of PE was higher at 0.48% (p = 0.045, chi-square). CONCLUSIONS: Increased use of PVCFs failed to decrease the overall rate of PE in our trauma patient population. PMID- 10472934 TI - Evolving technology for ultrasound localization and excision of nonpalpable breast cancer. PMID- 10472935 TI - James Bolton, MD (1812-1869): reflections on an American surgeon. PMID- 10472936 TI - Wire-guided transoral esophagogastrostomy for carcinoma of the cervical esophagus. PMID- 10472937 TI - Spontaneous pneumothorax from apical pulmonary blebs. PMID- 10472938 TI - Preoperative needle biopsy is helpful to the breast surgeon. PMID- 10472939 TI - Postoperative liver failure. PMID- 10472940 TI - Concomitant major hepatectomy and inferior vena cava reconstruction. PMID- 10472941 TI - Dear author--advice from a retiring editor. AB - This commentary, detailing the handling of a manuscript by the editor and guiding authors on preparing manuscripts and responding to reviews, provides parting advice to authors from a retiring editor. A close reading of this commentary will give some insight into the editorial process at the American Journal of Epidemiology through the observations of one of its editors. PMID- 10472942 TI - Glutathione S-transferase M1 genotypes and the risk of vulvar cancer: a population-based case-control study. AB - Glutathione S-transferases (GSTs) facilitate the excretion of a variety of potential carcinogens. Some 50-60% of Caucasians are homozygous for the null allele of GSTM1, a gene responsible for the presence of one of these enzymes. The authors examined whether women with the GSTM1 null genotype are at altered risk of vulvar cancer. They obtained peripheral blood specimens from 18- to 79-year old residents of King, Pierce, and Snohomish counties of western Washington who were diagnosed with vulvar cancer between April 1991 and June 1994. Blood specimens were also obtained from controls identified via random digit telephone dialing of western Washington households. The authors determined the GSTM1 genotype of 137 cases (120 in situ and 17 invasive cases) and 248 controls. The frequency of the GSTM1 null genotype was 46.7% among cases and 57.3% among controls. The age-adjusted odds ratio associated with the GSTM1 null genotype was 0.7 (95% confidence interval: 0.4, 1.0). Among current smokers of cigarettes, the age-adjusted odds ratio associated with the GSTM1 null genotype was 0.5 (95% confidence interval: 0.2, 0.9), differing little between heavy and light smokers. Our data suggest that women with the GSTM1 null genotype are not at increased risk of vulvar cancer. PMID- 10472943 TI - Mutagenic drinking water and risk of male esophageal cancer: a population-based case-control study. AB - Drinking mutagenic downstream water from the Huangpu River was hypothesized to have increased the risk for male esophageal cancer in Shanghai, China. The authors conducted a population-based case-control study of a total of 71 esophageal cancer deaths and 1,122 controls collected during a 5-year follow-up period, 1984-1988, from four male cohorts born before January 1, 1944, living in four communities consuming water with different mutagenicities in the Shanghai area. The controls represented a 1% random sample of the defined living cohorts selected at the end of each of the 5 years of follow-up. Logistic regression showed an odds ratio of 2.77 (95% confidence interval: 1.52, 5.03) for drinking mutagenic downstream water from the river versus drinking nonmutagenic upstream water after controlling for possible confounders including age, disease history (hepatitis, cirrhosis, schistosomiasis, digestive tract ulcer), hazardous occupational history, pesticide exposure, lifestyle factors (cigarette smoking, tea intake, and alcohol intake), dietary habits (intake of pickled vegetables, maize, peanuts, and cured meat), education, poverty, urban environment, and water chlorination. PMID- 10472944 TI - DDE and DDT in breast adipose tissue and risk of female breast cancer. AB - A case-control study was conducted in Connecticut from 1994 to 1997 to investigate the relation between dichlorodiphenyldichloroethane (DDE) and dichlorodiphenyltrichloroethane (DDT) exposure and breast cancer risk. Cases and controls were women aged 40-79 years, who had breast-related surgery at the Yale New Haven Hospital and from whose surgical specimen the authors could obtain at least 0.4 g of breast adipose tissue for chemical analyses. A total of 304 incident breast cancer cases (including 62 in situ carcinomas) and 186 benign breast disease controls were recruited into the study. Tissue levels of DDE and DDT were measured using gas chromatography. Statistical significance for comparisons of mean levels of DDE and DDT was calculated using analysis of variance and rank sum tests. A logistic regression model was used to estimate the association and to control confounding. The age-adjusted geometric mean tissue level of DDE for cases (736.5 ppb) was similar to that for the controls (784.1 ppb). DDT levels were also similar for cases (51.8 ppb) and controls (55.6 ppb). The adjusted odds ratio is 0.9 (95% confidence interval: 0.5, 1.5) for DDE and 0.8 (95% confidence interval: 0.5, 1.5) for DDT when the highest quartile was compared with the lowest. These results do not support an association between adipose tissue levels of DDE and DDT and breast cancer risk. PMID- 10472945 TI - Risk factors for basal cell carcinoma of the skin in men: results from the health professionals follow-up study. AB - The authors examined the relation of constitutional factors and sun exposure to risk of basal cell carcinoma of the skin (BCC) in a prospective cohort of 44,591 predominantly Caucasian US male health professionals, 40-75 years of age and free of cancer at enrollment in 1986. During 8 years of follow-up, 3,273 cases of self reported BCC were documented. The following variables were each associated with an elevated risk of BCC: having red hair; green, hazel, or blue eyes; a tendency to sunburn; and north European ancestry. The lifetime number of blistering sunburns was also positively associated with BCC risk (p trend < 0.0001). Compared with men who as teenagers had been outside less than once a week, men who had been outside weekly (relative risk (RR) = 1.30; 95% confidence interval (CI): 1.14, 1.47) and daily (RR = 1.42; 95% CI: 1.24, 1.63) had an elevated risk of BCC. Living in a region of residence with high solar radiation as an adult was also associated with an increased risk of BCC (RR = 1.48; 95% CI: 1.36, 1.60), whereas living in such a region only in childhood did not increase BCC risk. These results confirm the role of constitutional factors and suggest that adult sun exposure increases BCC risk. PMID- 10472946 TI - Random-effects meta-analyses are not always conservative. AB - It is widely held that random-effects summary effect estimates are more conservative than fixed-effects summaries in epidemiologic meta-analysis. This view is based on the fact that random-effects summaries have higher estimated variances and, consequently, wider confidence intervals than fixed-effects summaries when there is evidence of appreciable heterogeneity among the results from the individual studies. In such instances, however, the random-effects point estimates are not invariably closer to the null value nor are their p values invariably larger than those of fixed-effects summaries. Thus, random-effects summaries are not predictably conservative according to either of these two connotations of the term. The authors give an example from a meta-analysis of water chlorination and cancer in which the random-effects summaries are less conservative in both of these alternative senses and possibly more biased than the fixed-effects summaries. The discussion of when to use random effects and when to use fixed effects in computing summary estimates should be replaced by a discussion of whether summary estimates should be computed at all when the studies are not methodologically comparable, when their results are discernibly heterogeneous, or when there is evidence of publication bias. PMID- 10472947 TI - Use of cimetidine, omeprazole, and ranitidine in pregnant women and pregnancy outcomes. AB - Evidence documenting the safety of acid-suppressing drugs in pregnancy is very limited. The authors assessed the prevalence of congenital malformations in first trimester-exposed pregnancies to cimetidine, omeprazole, and ranitidine and compared it with nonexposed pregnancies between 1991 and 1996. Two different sources were used, the United Kingdom General Practice Research Database and the Italian Friuli-Venezia Giulia Health Database. The final study cohort included 1,179 pregnancies from the United Kingdom and 1,057 from Italy. Abortions or ectopic pregnancies were not included. There were 20 stillbirths and 2,261 live born babies in both cohorts combined, with 100 offspring identified with a malformation. The overall malformation rate was 4.4%. The relative risks for nongenetic congenital malformations associated with the use of cimetidine, omeprazole, and ranitidine were 1.2 (95% confidence interval (CI): 0.6, 2.3), 0.9 (95% CI: 0.3, 2.2), and 1.4 (95% CI: 0.8, 2.4), respectively, compared with the nonexposed. No specific grouping in the distribution of malformations was observed in any of the three exposed groups. Moreover, no relation was found between drug exposure and preterm delivery or growth retardation. These findings suggest that the use of acid-suppressing drugs during the first trimester of pregnancy is not associated with a major teratogenic risk. PMID- 10472948 TI - Eosinophilia and positive skin tests predict cardiovascular mortality in a general population sample followed for 30 years. AB - The authors investigated whether two objective allergy markers, peripheral blood eosinophilia and skin tests for common aeroallergens, were associated with cardiovascular death. Of 5,382 subjects in the Vlagtwedde-Vlaardingen Study (the Netherlands) with data on allergy markers in 1965-1972, 507 subjects died from cardiovascular disease during 30 years of follow-up. Subjects with eosinophilia had an increased risk of cardiovascular death (relative risk (RR) = 1.7; 95% confidence interval (CI): 1.4, 2.2), including ischemic heart disease death (RR = 1.6; 95% CI: 1.2, 2.2) and cerebrovascular death (RR = 2.3; 95% CI: 1.4, 3.8), independent of major risk factors. This association was limited to subjects with a percentage of the predicted forced expiratory volume in 1 second (FEV1 % predicted) of <100%. Positive skin tests were associated with a significantly reduced cardiovascular mortality in subjects with normal lung function and weight who did not smoke (RR = 0.15; 95% CI: 0.05, 0.46). Conversely, when subjects with positive skin tests had a body mass index of > or =25 kg/m2, had an FEV1 % predicted of <80%, or smoked, they had an increased risk for cardiovascular mortality. These results were not restricted to asthmatics. Our data suggest a possible link between eosinophilia and positive skin tests and cardiovascular mortality, especially in combination with other risk factors associated with its mortality. PMID- 10472949 TI - Plasma fatty acid composition and 6-year incidence of hypertension in middle-aged adults: the Atherosclerosis Risk in Communities (ARIC) Study. AB - The association of baseline fatty acid composition in plasma cholesterol esters with 6-year incidence of hypertension was examined in middle-aged Minneapolis participants of the Atherosclerosis Risk in Communities (ARIC) Study (1987-1995). Compared with those who were never hypertensive (n = 1,975), incident hypertensives (n = 413) had statistically significantly higher baseline levels of palmitic (16:0) and palmitoleic (16:1n7) acids but lower levels of linoleic (18:2n6) acid and the polyunsaturated/saturated fatty acids ratio (P/S ratio). Among polyunsaturated fatty acids, levels of dihomo-gamma-linolenic (20:3n6) and arachidonic (20:4n6) acids were statistically significantly higher in incident hypertensives, compared with normotensives. After adjustment for age, sex, body mass index, waist/hip ratio, smoking status, ethanol intake, education level, physical activity, and baseline systolic blood pressure in separate models, the odds ratio estimates of incident hypertension for an interquartile increment of a fatty acid in cholesterol esters were 1.26 (95% confidence interval (CI): 1.05, 1.51) for 16:0, 1.11 (95% CI: 0.96, 1.28) for 16:1n7, 1.01 (95% CI: 0.85, 1.21) for 20:3n6, 1.14 (95% CI: 1.03, 1.27) for 20:5n3, 0.81 (95% CI: 0.68, 0.96) for 18:2n6, and 0.83 (95% CI: 0.70, 0.99) for the P/S ratio. The authors conclude that reduced levels of linoleic acid and the P/S ratio and elevated levels of palmitic and arachidonic acids are associated with a higher risk of hypertension. PMID- 10472950 TI - Factors associated with functional decline and improvement in a very elderly community-dwelling population. AB - The objective of this study was to analyze the factors associated with functional decline and improvement in a community-dwelling population of people aged 75 years and older. A representative sample of elderly people living at home in the city of Sherbrooke (Quebec, Canada) was assessed yearly on three occasions (1991 1993) by a nurse. A health questionnaire, together with standardized instruments measuring disabilities, cognitive status, and depressive mood, was administered on the three assessments. From the 655 subjects who agreed to participate, a total of 504 subjects completed the study. The most important factors associated with functional decline were the number of days off regular activities (odds ratio (OR) = 1.31), the number of hot meals per day (OR = 1.59), and cognitive status (OR = 0.96), whereas weight loss (OR = 0.37) and living alone (OR = 0.54) were significant protective factors. Previous declines in functional autonomy, cognitive state, or mood were not independent risk factors. The most important factors associated with functional improvement were the disability score (OR = 1.08), previous functional decline (OR = 6.06), and decline in the disability score (OR = 1.09). Perceiving health to be worse than it was the previous year was a significant factor against improvement (OR = 0.24). This study identifies risk factors that can be helpful for targeting high-risk subgroups within the very elderly population who may benefit from a preventive program. PMID- 10472951 TI - Validity of self-reports of fractures in perimenopausal women. AB - The validity of self-report of fractures in postal inquiry among perimenopausal women was evaluated. Self-reports of fractures in the 1989 baseline postal inquiry data of the Kuopio Osteoporosis Risk Factor and Prevention Study (OSTPRE) were compared with information in patient records. The study population consisted of 373 women who reported fractures sustained during the last 10 years and 200 randomly selected women who did not report fractures from a population base of 2,007 women aged 47-56 years. Self-report as a screening test for fracture was evaluated in the total sample of 2,007 women by estimating the number of false negative reports in all the women who did not report a fracture with the information on these 200 women. Of the self-reports of fractures, 84% proved to be true fractures, 12% soft tissue injuries, and the rest either self-diagnoses or misnomers. Self-report of wrist fracture was more accurate (95%). The sensitivity of self-report to detect fracture was 78% for all fractures and 95% for wrist fracture, while the respective specificities were 96 and 99%. Self report is a relatively accurate way to obtain information about past major fractures in perimenopausal women. However, it is rather insensitive in the detection of minor fractures, if the reporting period is several years. PMID- 10472952 TI - Early childhood mortality from community-acquired infections. AB - In this study, the authors identified maternal and child characteristics that were independent predictors of death from infectious diseases acquired in the community and determined if these factors could be used to identify groups of children with excess risk of mortality from infection. A historical cohort study was conducted of children less than 5 years of age between 1985 and 1994 (the study period), who were born in Tennessee, and had complete information on their birth certificates. The primary outcome was death from infection identified from death certificates and confirmed through medical record review. Among the 1,014,976 children less than 5 years of age, who contributed 3,351,568 child years of follow-up, there were 247 deaths from infections (7.4 deaths from infections per 100,000 child-years). Respiratory infections accounted for approximately one half of the deaths. Children having three or more older siblings or birth weight of less than 1,500 g had a 3-fold and 10-fold increased risk of death from infection, respectively, while children with both characteristics had a nearly 20-fold increased risk that persisted beyond the first year of life. Interventions should be focused on prevention of these infections in vulnerable children. At-risk children should be targeted for careful follow-up and early hospitalization when signs of infection develop. PMID- 10472953 TI - Maternal smoking and childhood asthma. AB - The role of maternal smoking as a causal factor for the incidence of childhood asthma is still not clearly established. It was investigated among 3- and 4-year old-child incident cases confirmed by a 6-year follow-up (n = 294) and cases who no longer had symptoms after diagnosis (n = 110). The study took place in Montreal, Canada, between 1988 and 1997. Persistent and transient cases were compared with their respective controls from the original case-control study of incidence. The odds ratio for heavy maternal smoking adjusted for known risk factors for asthma was 3.84 (95% confidence interval: 1.68, 8.76) among persistent cases and close to one among transient cases. PMID- 10472954 TI - The postwar hospitalization experience of Gulf War Veterans possibly exposed to chemical munitions destruction at Khamisiyah, Iraq. AB - Using Department of Defense hospital data, the authors examined the postwar hospitalization experience from March 1991 through September 1995 of US Gulf War veterans who were near Khamisiyah, Iraq, during nerve agent munition destruction in March 1991. Multiple sources of meteorologic, munition, and toxicology data were used to circumscribe geographic areas of low level, vaporized nerve agent for 4 days after the destruction. Plume estimates were overlaid on military unit positions, and exposure was estimated for the 349,291 US Army Gulf War veterans. Exposure was classified as not exposed (n = 224,804), uncertain low dose exposure (n = 75,717), and specific estimated subclinical exposure (n = 48,770) categorized into three groups for dose-response evaluation. Using Cox proportional hazard modeling, the authors compared the postwar experiences of these exposure groups for hospitalization due to any cause, for diagnoses in 15 unique categories, and for specific diagnoses an expert panel proposed as most likely to reflect latent disease from such subclinical exposure. There was little evidence that veterans possibly exposed to the nerve agent plumes experienced unusual postwar morbidity. While there were several differences in hospitalization risk, none of the models suggested a dose-response relation or neurologic sequelae. These data, having a number of limitations, do not support the hypothesis that Gulf War veterans are suffering postwar morbidity from subclinical nerve agent exposure. PMID- 10472955 TI - Re: "Cigarette smoking and the colorectal adenoma-carcinoma sequence: a hypothesis to explain the paradox". PMID- 10472956 TI - Re: "Population attributable fraction estimation for established breast cancer risk factors: considering the issues of high prevalence and unmodifiability". PMID- 10472957 TI - The changing nature of glaucoma. PMID- 10472959 TI - Glaucoma screening: the importance of combining test data. AB - The objective of this study was to evaluate the effectiveness of screening tests for primary open angle glaucoma, both singly and in combination, using a decision analysis approach. A range of screening tests were carried out on 145 nonglaucomatous patients and 67 cases of previously undiagnosed glaucoma. Receiver operator characteristic curves were constructed for single test data to show the trade-off between sensitivity and specificity for varying cut-off criteria. The best discriminators of glaucoma were, in rank order: (1) a multiple stimulus static visual field screening test, (2) optic disc cupping, and (3) intraocular pressure. Decision curves were also constructed for various combinations of screening tests, where the inclusion of the tests were based on discriminant analyses. Sensitivities and specificities of more than 0.90 were obtained when visual field screening, optic disc cupping, and intraocular pressure were combined. Data from other tests when combined with these three variables failed to provide a significant improvement in discrimination. PMID- 10472958 TI - New technologies for diagnosing and monitoring glaucomatous optic neuropathy. AB - BACKGROUND: Recently, instruments have been developed to provide real-time, quantitative measurements of the optic disc and retinal nerve fiber layer (RNFL) for use in glaucoma management. Our objective is to (1) provide an overview of two of these instruments, the confocal scanning laser ophthalmoscope (Heidelberg Retina Tomograph, HRT) and scanning laser polarimeter (Nerve Fiber Analyzer, NFA) and (2) compare measurements obtained with these instruments to clinical features used in the diagnosis of glaucoma. METHODS: Twenty glaucoma patients, 4 normal subjects and 20 glaucoma subjects were included. All subjects had images obtained with the HRT and NFA, and RNFL and optic disc photography completed within 5 weeks of each other. The HRT results were compared with qualitative evaluation of stereophotographs of the optic disc, and NFA results were compared against a semi quantitative RNFL photograph severity score. RESULTS: Twenty-five (57%) subjects had thinning of the neuroretinal rim identified by evaluation of stereoscopic optic disc photographs. Despite overlap, HRT measurements of rim volume, rim area, and rim/disc ratio were significantly smaller in eyes with evidence of rim thinning than in eyes with no evidence of rim thinning. Moderate to severe RNFL damage was detected by evaluation of photographs in 25 (57%) of subjects. NFA RNFL thickness measures were smaller in eyes with moderate to severe RNFL damage than in relatively healthy eyes. CONCLUSIONS: Previous studies have documented the reproducibility of these instruments and suggested analytic techniques for improving their ability to differentiate between normal and glaucoma eyes. Our results indicate that despite overlap in values, these instruments provide measurements that reflect clinically relevant features of the optic disc and RNFL. Whether these technologies can improve our ability to detect glaucomatous progression over time needs to be determined with well-designed longitudinal studies and comparison with established diagnostic techniques for evaluating glaucomatous optic neuropathy. PMID- 10472960 TI - Iris transillumination defects in the pigment dispersion syndrome as detected with infrared videography: a comparison between a group of blacks and a group of nonblacks. AB - BACKGROUND: Infrared image analysis has been reported to be the most sensitive method to detect iris transillumination defects (ITDs) associated with the pigment dispersion syndrome (PDS). Although a common sign of PDS in patients with lighter irides, ITDs that are demonstrated with standard slit lamp examination are frequently absent in blacks with the disease. This absence may lead to confusion in diagnosis; therefore improved methods of evaluation could be beneficial. Although infrared image analysis has been investigated in nonblacks, it has not been applied to a group of blacks with the condition. METHODS: Six blacks (11 eyes, all irides brown) with PDS and eight nonblacks (15 eyes, 10 blue irides, 5 brown irides) with PDS underwent analysis. ITDs were searched for with traditional slit lamp examination and with infrared videography. RESULTS: Among the blacks, 2 of 11 eyes (18%) showed ITDs via infrared analysis, a frequency that was significantly lower (p < 0.001) than the frequency exhibited among the nonblacks (100%) when examined with the same technique. Standard slit lamp examination also showed a significantly lower frequency of ITDs among the blacks compared to the nonblacks (p < 0.001). Infrared videography detected ITDs at least one grade higher in 1 of 11 eyes (9%) among the black group and in 7 of the 15 eyes (47%) among the nonblacks. Using the Wilcoxon signed ranks test, the difference in the ability of infrared videography to detect and rank the iris defects in the nonblack PDS group was significantly greater when compared to standard slit lamp examination (p = 0.011), but infrared analysis did not increase detection or change the mean ranking of those in the black group (p = 0.317). CONCLUSION: In this group of patients, infrared videography did not significantly improve the detection and visibility of ITDs in blacks, as it did in the nonblacks. The frequency that ITDs were detected with standard slit lamp examination and with infrared videography was significantly greater in the group of nonblacks compared to the blacks who exhibited PDS. PMID- 10472961 TI - Random dot motion stimuli are more sensitive than light stimuli for detection of visual field loss in ocular hypertension patients. AB - PURPOSE: To determine whether motion detection perimetry or luminance size threshold perimetry (a test using the motion perimetry method with luminance stimuli) is more sensitive in detecting visual loss in ocular hypertension patients. METHODS: Motion perimetry uses a customized computer graphics program to detect a subject's ability to identify a coherent shift in position of moving dots in a defined circular area against a background of fixed dots. Motion size threshold is defined as the smallest circular area within which dot motion is detected. Patients respond by touching the area of the computer monitor where they perceive the stimulus with a light pen. The localization errors are measured as the number of pixels from target center for each trial. Luminance size threshold perimetry uses the same technique except the background is dark gray and the stimuli are filled lighter gray circles. We tested one eye in each of 27 ocular hypertension patients and 27 age-matched normal subjects with both tests. Our main outcome measures were motion and luminance size thresholds, total deviation probability plot data, and spatial localization errors. RESULTS: With the total deviation probability plot analysis, the ocular hypertension patients had a greater number of abnormal test locations with motion perimetry stimuli than with luminance stimuli. The abnormal test points were located most often in the superior and inferior nasal regions. Six subjects had nerve fiber bundle-like defects to motion stimuli whereas three patients had defects with luminance size threshold perimetry. The ocular hypertension patients had significantly greater localization errors than the controls with both tests. CONCLUSIONS: Using a size thresholding technique in ocular hypertension patients, random dot motion stimuli appear to be more sensitive than luminance stimuli. Errors in stimulus localization are significantly increased in ocular hypertension patients, independent of the stimulus (motion or luminance) used. PMID- 10472962 TI - Progressive visual field defects from experimental glaucoma: measurements with white and colored stimuli. AB - PURPOSE: Our purpose was to study the effects of using monochromatic test stimuli to measure the relative rate of progression of visual field defects caused by experimental glaucoma. METHODS: Visual field measurements were obtained by static perimetry from trained macaque monkeys with laser-induced, unilateral glaucoma. The visual field defects were assessed by perimetric (global) indices derived from comparisons of experimental visual fields to the expected normal fields of monkeys. Three types of perimetry stimuli were used, the conventional white Goldmann III and two monochromatic (460 and 620 nm) Goldmann V test stimuli. The relationships between field defects with white and monochromatic stimuli were investigated by linear regression of the Z-scores for the perimetric indices. RESULTS: The correlations between the mean deviation global indices for chromatic vs. white stimuli were high (r > 0.9) and linear throughout the period of progression of field defects. The slopes of the regression lines typically were greater than unity, indicating that statistical significance was higher for visual field defects measured with chromatic stimuli than with white light stimuli. The higher significance level for defects measured with chromatic stimuli was not explained by a difference in visual thresholds, because the thresholds with chromatic and white light were highly correlated across the full range of visual field defects, from initial-onset to end-state. This result also suggests that the early detection of glaucomatous visual defects with monochromatic stimuli does not reflect a selective loss of retinal ganglion cells. CONCLUSIONS: Although these experiments do not suggest an alternative neural mechanism for the clinical utility of perimetry with chromatic light for the early detection of glaucoma, it is very likely that the combinations of neural and/or analytical factors that explain the utility of perimetry with chromatic stimuli will also provide an explanation for the higher sensitivities in identifying early glaucoma reported for other prototype stimuli. PMID- 10472963 TI - Frequency doubling technology perimetry using a 24--2 stimulus presentation pattern. AB - PURPOSE: To assess whether smaller targets and a 24-2 stimulus presentation pattern would improve the ability of frequency doubling technology (FDT) perimetry to detect and characterize early glaucomatous visual field loss. METHODS: One hundred normal subjects between the ages of 20 and 85 participated in this study. In addition, 53 patients who either had early glaucomatous visual field loss (n = 23) or were high-risk glaucoma suspects with normal conventional visual fields (n = 30) were evaluated with the commercial version of FDT perimetry (full threshold test) with 17 stimuli (four 10 degrees diameter square targets per quadrant and a central 5 degrees circular target) and a custom version of FDT perimetry using 54 stimuli (4 degrees targets with 6 degrees grid spacing) arranged in a 24-2 stimulus presentation pattern. RESULTS: The custom FDT test using a 24-2 stimulus presentation pattern had a similar dynamic range, and demonstrated normal aging characteristics and test-retest reliability that were similar to the commercial version of FDT perimetry using 17 larger stimuli. Both FDT tests showed an age-related sensitivity reduction of approximately 0.6 dB per decade, and exhibited an average test-retest reliability of 1 to 1.5 dB. The custom 24-2 FDT perimetry test had a greater variation of sensitivity with eccentricity than the commercial version of FDT perimetry that was probably related to the difference in stimulus size. The custom 24-2 FDT perimetry test had a greater percentage of abnormal test locations than the commercial FDT test for both early glaucomas and high-risk glaucoma suspects. CONCLUSIONS: FDT perimetry can be performed with smaller targets using a presentation pattern that is similar to conventional automated perimetry. In comparison to the commercially available 17 target display, the 24-2 stimulus pattern appears to have modestly higher sensitivity for detection of early glaucomatous loss and provides better characterization of the pattern of visual field loss, but the test takes approximately twice as long. PMID- 10472964 TI - Normal aging effects for frequency doubling technology perimetry. AB - PURPOSE: To determine the influence of normal aging on contrast sensitivity for frequency doubling technology (FDT) perimetry. METHODS: Contrast sensitivity measures were obtained for frequency-doubled stimuli (0.25 cycles per degree sinusoidal gratings undergoing 25 Hz counterphase flicker) at 17 target locations (4 per quadrant plus the central 5 degrees ) using a prototype of the Welch Allyn (Skaneateles, NY)/Humphrey Systems FDT perimeter (Humphrey Systems, Dublin, CA). A total of 407 normal subjects (761 eyes) between the ages of 15 and 85 years were tested. RESULTS: Between the ages of 15 and 60 years there was an approximately linear decrease in contrast sensitivity of 0.6 dB per decade. After the age of 70, there was a slightly greater sensitivity loss with age. There were no meaningful differences in sensitivity loss as a function of age for different visual field locations. A small but consistent reduction in contrast sensitivity (approximately 0.7 dB) was found at all visual field locations for the second eye tested that may be due to a central adaptation process. CONCLUSIONS: Normal aging effects for FDT perimetry are similar to those obtained for conventional automated perimetry, except that the FDT perimetry aging effects do not appear to be eccentricity dependent. These normative data provide a basis for establishing a statistical analysis procedure and probability plots for FDT perimetry. PMID- 10472965 TI - A new look at threshold estimation algorithms for automated static perimetry. AB - Automated perimetry is often associated with lengthy test times when a staircase algorithm is applied. This arises because the fixed step sizes used during threshold estimation (e.g.,4/2 dB) yield reduced test efficiency, with test times being dependent on the relative positioning of the start and endpoints, as well as the step size. Neighborhood logic may speed up the process, although several presentations are still required for normal threshold values and many more presentations are required for abnormal values. We consider whether there is any justification for using a fixed step size during the threshold procedure. We show how empirical data can be applied, within a Bayesian framework, to reduce test time with little or no loss of accuracy. Finally, we demonstrate the effect that the starting probability density function can have on test efficiency by implementing an empirically determined and bimodal probability density function that provides fast outcomes. PMID- 10472966 TI - Normal-tension glaucoma and central corneal thickness. AB - PURPOSE: To determine whether a decreased corneal thickness is a primary factor in the lower intraocular pressure readings found in normal-tension glaucoma patients. METHODS: Fourteen patients with normal-tension glaucoma were compared with 14 age- and sex-matched glaucoma patients. Corneal thickness was measured by ultrasonic pachymetry on each eye. RESULTS: The corneal thickness (mean +/- SD) in patients with normal-tension glaucoma was 0.536 +/- 0.035 mm. The corneal thickness in patients with glaucoma was 0.537 +/- 0.035 mm. The 95% confidence interval for the difference between the two groups was -0.027 to +0.027 mm. CONCLUSION: It has been documented that increased corneal thickness leads to high readings of intraocular pressure. Therefore, decreased corneal thickness may lead to a lower intraocular pressure reading. This study suggests that for most normal tension glaucoma patients corneal thickness is not a major factor in accounting for the lower intraocular pressure measurements when compared with primary open angle glaucoma patients. PMID- 10472967 TI - Fundamentals of exact perimetry. 1945. PMID- 10472968 TI - The "third-best" strategy for treating head and neck cancer. PMID- 10472969 TI - MR perfusion imaging. PMID- 10472970 TI - Imaging brain abscesses with diffusion-weighted and other sequences. PMID- 10472971 TI - Intraarterial thrombolysis for cerebral infarction: to treat or not to treat, and how? PMID- 10472972 TI - Comparison of three-dimensional visualization techniques for depicting the scala vestibuli and scala tympani of the cochlea by using high-resolution MR imaging. AB - BACKGROUND AND PURPOSE: Cochlear implantation requires introduction of a stimulating electrode array into the scala vestibuli or scala tympani. Although these structures can be separately identified on many high-resolution scans, it is often difficult to ascertain whether these channels are patent throughout their length. The aim of this study was to determine whether an optimized combination of an imaging protocol and a visualization technique allows routine 3D rendering of the scala vestibuli and scala tympani. METHODS: A submillimeter T2 fast spin-echo imaging sequence was designed to optimize the performance of 3D visualization methods. The spatial resolution was determined experimentally using primary images and 3D surface and volume renderings from eight healthy subjects. These data were used to develop the imaging sequence and to compare the quality and signal-to-noise dependency of four data visualization algorithms: maximum intensity projection, ray casting with transparent voxels, ray casting with opaque voxels, and isosurface rendering. The ability of these methods to produce 3D renderings of the scala tympani and scala vestibuli was also examined. The imaging technique was used in five patients with sensorineural deafness. RESULTS: Visualization techniques produced optimal results in combination with an isotropic volume imaging sequence. Clinicians preferred the isosurface-rendered images to other 3D visualizations. Both isosurface and ray casting displayed the scala vestibuli and scala tympani throughout their length. Abnormalities were shown in three patients, and in one of these, a focal occlusion of the scala tympani was confirmed at surgery. CONCLUSION: Three-dimensional images of the scala vestibuli and scala tympani can be routinely produced. The combination of an MR sequence optimized for use with isosurface rendering or ray-casting algorithms can produce 3D images with greater spatial resolution and anatomic detail than has been possible previously. PMID- 10472973 TI - Assessment of internal auditory canal tumors: a comparison of contrast-enhanced T1-weighted and steady-state T2-weighted gradient-echo MR imaging. AB - BACKGROUND AND PURPOSE: Although contrast-enhanced T1-weighted MR imaging is the standard of reference for diagnosing tumor in the cerebellopontine angle, high resolution T2-weighted imaging may show more details of the seventh and eighth cranial nerve branches, resulting in more accurate tumor volume measurements. The purpose of this study was to compare two MR sequences for their ability to delineate internal auditory canal tumors. METHODS: Twenty-seven ears in 21 patients with 16 confirmed schwannomas were studied with the 3D T2-weighted prototype segment-interleaved motion-compensated acquisition in steady state (SIMCAST) and the T1-weighted contrast-enhanced spoiled gradient-echo (SPGR) techniques. Twenty-eight axial sections were acquired using parameters of 17/3.3 (TR/TE), a 40 degrees flip angle, a 20 x 15-cm or 22 x 16-cm field of view (FOV), a 512 x 256 matrix, and a 0.4- or 1.2-mm section thickness for the SIMCAST technique, and 30/4.2, a 30 degrees flip angle, a 20 x 20-cm FOV, a 512 x 288 matrix, and a 1.5-mm section thickness for the SPGR technique. Tumor appearance and depiction of surrounding anatomy, including the cranial nerves, were evaluated. Tumor volumes were measured by manual tracing. RESULTS: Both sequences clearly identified tumors that ranged in size from 0.06 to 3.0 cm3. Measurements on both sequences agreed, on average, within 14%. The information from both sequences was complementary. SIMCAST usually delineated the CSF spaces better, whereas SPGR more clearly showed the tumor/brain boundary. CONCLUSION: SIMCAST and SPGR are suitable for tumor detection and volume measurements. SPGR has somewhat better contrast, but SIMCAST excels at depicting the surrounding anatomy and tumor involvement of the seventh and eighth cranial nerves. PMID- 10472974 TI - Thallium-201 single-photon emission CT versus CT for the detection of recurrent squamous cell carcinoma of the head and neck. AB - BACKGROUND AND PURPOSE: Thallium-201 single-photon emission computed tomography (SPECT) can be used to detect primary squamous cell carcinoma (SCCA) of the head and neck. Nevertheless, there have very few studies performed to evaluate the ability of thallium-201 to depict recurrent SCCA. The purpose of this study was to compare the ability of thallium-201 SPECT with CT to enable detection of recurrent SCCA of the upper aerodigestive tract. METHODS: Thirty-three patients with a history of previously treated SCCA of the extracranial head and neck underwent thallium-201 SPECT imaging and contrast-enhanced CT. A neuroradiologist and nuclear medicine physician with knowledge of the primary site evaluated all thallium-201 studies for abnormal radiotracer uptake at the primary site. These results were correlated with histologic findings and clinical follow-up in all patients. All patients were followed up for a minimum of 2 years after completion of treatment. The McNemar test was used to determine statistical significance. RESULTS: The diagnostic accuracy of thallium-201 SPECT was as follows: sensitivity, 88%; specificity, 94%; positive predictive value, 93%; and negative predictive value, 89%. The diagnostic accuracy of CT was as follows: sensitivity, 100%; specificity, 24%; positive predictive value, 55%; and negative predictive value, 100%. The diagnostic accuracy of thallium was superior to CT (P = .01). CONCLUSION: Thallium-201 SPECT is superior to CT for differentiating recurrent tumor from post-treatment changes and may complement CT in the evaluation of previously treated SCCA of the extracranial head and neck. PMID- 10472975 TI - Osteometry of the mandible performed using dental MR imaging. AB - BACKGROUND AND PURPOSE: On cross-sectional and panoramic reformatted images from axial (dental) CT scans of the mandible it may be difficult to identify the inferior alveolar neurovascular bundle (IANB) in patients lacking a clear-cut bony delimitation of the mandibular canal. Dental MR images are comparable to dental CT scans, which directly show the IANB; however, measurements of length may not be reliable owing to susceptibility artifacts and field inhomogeneities in the oral cavity. Therefore, the accuracy of length measurements on dental MR images was compared with that on dental CT scans and direct osteometry. METHODS: Dental T1-weighted MR imaging using a high-resolution turbo gradient-echo sequence and dental CT were performed in six anatomic specimens. The axial scans were reformatted as panoramic and cross-sectional reconstructions on a workstation and characteristic cross sections were obtained from all mandibles. The longest axis in the bucco-lingual and apico-basal directions, the distances from the top of the mandibular canal to the top of the alveolar ridge and from the bottom of the mandibular canal to the base of the mandible, and the diameter of the bone cortex at the alveolar ridge were measured with direct osteometry on the cross sections and compared with measurements on corresponding MR and CT reformatted images. RESULTS: The correlation between direct osteometry and dental MR and CT was strong, except for the bone cortex diameter at the top of the alveolar ridge, where only a moderate correlation was found. Means of comparable length measurements were not significantly different among the three methods. CONCLUSION: The accuracy of length measurements in the jaw bones obtained using dental MR is comparable to that of dental CT and is not significantly different from direct osteometry. Thus, dental MR is a potential alternative to CT for dental imaging. PMID- 10472976 TI - Optic nerve enlargement in Krabbe's disease. AB - We report imaging and gross pathologic findings from two cases of Krabbe disease in which there was marked enlargement of the intracranial optic nerves. Numerous globoid cells were observed in the optic nerves at autopsy in one case. Krabbe disease should be included in the differential diagnosis of children with enlargement of the optic nerves. PMID- 10472977 TI - Cerebral blood flow-related signal changes during breath-holding. AB - BACKGROUND AND PURPOSE: In the past, functional MR imaging techniques have been used successfully to determine cerebrovascular reactivity (CVR) to various stimuli, complementing the arsenal of functional brain investigations feasible with MR imaging. While previous studies have focused on blood oxygenation changes under vasodilatory stress, the aim of this study was to assess regional cerebral blood flow (rCBF) changes during breath-holding by using a flow-sensitive alternating inversion recovery (FAIR) imaging technique. METHODS: In six healthy volunteers, FAIR images were acquired during alternating periods of breath holding and breathing at 40-second intervals after inspiration and at 30-second intervals after expiration, for a total dynamic scanning time of 10 minutes. To quantify the rCBF changes, we obtained 2.5-minute baseline samples during normal breathing. RESULTS: Repeated challenges of breath-holding induced an overall rise in rCBF. In general, rCBF changes were greatest in gray matter and were insignificant in white matter. Using the mean values of the baseline images collected before breath-holding to calculate the rCBF changes, we found that quantitative analysis yielded an rCBF increase of 47% to 87% after breath holding. The rCBF changes clearly depended on the breath-holding duration and technique; however, for one given breath-holding paradigm the results showed relatively small interindividual variability. CONCLUSION: rCBF changes during a simple vascular challenge can be detected and quantified by means of functional MR imaging at 1.5 T. Noninvasive assessment of CVR could become a useful clinical tool to identify persons with impaired CVR. PMID- 10472978 TI - MR high-resolution blood oxygenation level-dependent venography of occult (low flow) vascular lesions. AB - A new technique for detecting vascular malformations, high-resolution BOLD venography (HRBV), is described. This technique relies on the BOLD principle for detecting deoxygenated blood in low-flow malformations. HRBV images are acquired using a modified 3D gradient-echo with voxel volumes of 0.5 x 0.5 x 2 mm3. The magnitude data are masked with the phase images to enhance visibility of the venous structures and are displayed using the minimum intensity projection. Preliminary results for 10 patients show that HRBV is more sensitive in detecting cavernomas than is T2-weighted imaging, and lesions that are presumed to be telangiectasias are detected only with this technique. PMID- 10472979 TI - Multiple varices in the unilateral cerebral venous system. AB - A case of multiple cerebral varices located in the superficial cerebral veins and ipsilateral internal jugular vein is reported. PMID- 10472980 TI - Superficial siderosis of the CNS associated with multiple cavernous malformations. AB - Superficial siderosis of the CNS due to chronic, recurrent subarachnoid hemorrhage is an uncommon and potentially debilitating disorder. The classic clinical manifestation is progressive bilateral sensorineural hearing loss (SNHL), although ataxia and pyramidal signs also are observed frequently. Cavernous malformations rarely present with subarachnoid hemorrhage. We describe an unusual case of a young patient who presented with progressive, bilateral SNHL who was found to have superficial CNS siderosis associated with multiple cavernous malformations. PMID- 10472981 TI - Regional cerebral glucose metabolism and blood flow in a patient with Marchiafava Bignami disease. AB - We report functional neuroimaging studies of a 54-year-old man with Marchiafava Bignami disease (MBD). Glucose metabolic images obtained by [18F]-2-fluoro-2 deoxy-D-glucose positron emission tomography showed diffusely reduced whole brain metabolism and strongly decreased metabolism in the frontal and parietal lobes, orbital gyrus, and thalamus. Cerebral perfusion images showed a similarly decreased radioactivity pattern as the metabolic images. Functional neuroimages would be useful for understanding the pathophysiologic processes of MBD. PMID- 10472982 TI - Use of diffusion-weighted MR imaging in differential diagnosis between intracerebral necrotic tumors and cerebral abscesses. AB - The differential diagnosis between intracerebral necrotic tumors and cerebral abscesses is frequently impossible with conventional MR imaging. We report two cases of cerebral abscesses that showed high signal on diffusion-weighted echo planar imaging and a strongly reduced apparent diffusion coefficient. This appearance was not present in our cases of necrotic/cystic gliomas (eight cases) and necrotic metastases (two cases). We believe that diffusion-weighted MR imaging may be a diagnostic clue in cases of cerebral "ring-enhancing" masses. PMID- 10472983 TI - Analysis of cystic intracranial lesions performed with fluid-attenuated inversion recovery MR imaging. AB - BACKGROUND AND PURPOSE: T1-, T2-, and proton density (PD)-weighted sequences are used to characterize the content of cystic intracranial lesions. Fluid-attenuated inversion recovery (FLAIR) MR sequences produce T2-weighted images with water signal saturation. Therefore, we attempted to verify whether FLAIR, as compared with conventional techniques, improves the distinction between intracranial cysts with a free water-like content versus those filled with a non-free water-like substance and, consequently, aids in the identification of these lesions as either neoplastic/inflammatory or maldevelopmental/porencephalic. METHODS: Forty five cystic intracranial lesions were studied using T1-weighted, T2-weighted, FLAIR, and PD-weighted sequences. By means of clustering analysis of the ratio in signal intensity between the cystic intracranial lesions and CSF, the intracranial lesions were classified as filled with a free water-like content or with a non-free water-like substance. The results were compared with their true content as evaluated either histologically or on the basis of clinical, neuroradiologic, and follow-up features (necrotic material, 13 cases; accumulation of intercellular proteinaceous/myxoid material, eight cases; keratin, five cases; CSF, 19 cases). Cystic intracranial lesions were divided into two clinical groups, neoplastic/inflammatory and maldevelopmental/porencephalic, to evaluate the level of accuracy of each MR technique. The difference in absolute value signal intensity between CSF and cystic intracranial lesion content was calculated on FLAIR and PD-weighted images. RESULTS: PD-weighted and FLAIR sequences, unlike T1- and T2-weighted sequences, accurately depicted all cystic intracranial lesions containing necrotic or myxoid/proteinaceous intercellular material (non-free water-like) and most CSF-containing cystic intracranial lesions (free water-like). All imaging techniques inaccurately showed some of the keratin-containing cystic intracranial lesions and pineal cysts. The overall error rate was 22% for T1-weighted, 27% for T2-weighted, 9% for FLAIR, and 13% for PD-weighted sequences. The signal intensity difference between CSF and cystic intracranial lesion content was higher with FLAIR imaging. CONCLUSIONS: FLAIR imaging depicts far more accurately the content of cystic intracranial lesions and better reveals the distinction between maldevelopmental/porencephalic and neoplastic/inflammatory lesions than do conventional sequences. FLAIR has the added advantage of a higher signal intensity difference between cystic intracranial lesions and CSF. PMID- 10472984 TI - A longitudinal MR study of the presymptomatic phase in a patient with clinically definite multiple sclerosis. AB - We describe the dynamics and the nature of the presymptomatic phase of multiple sclerosis (MS) in a patient for whom MR abnormalities suggestive of MS were found before the development of clinical symptoms. The patient was monitored with serial monthly MR imaging of the brain and spinal cord for 5 months. Disease activity during the presymptomatic phase showed imaging characteristics comparable to that of early relapsing-remitting MS in terms of enhancing lesions, duration of enhancement, and new lesions depicted by T2-weighted imaging. Measurements derived from magnetization transfer imaging suggested that the amount and degree of tissue destruction within and outside the lesions revealed by T2-weighted imaging were mild. This, together with the fact that only one of the 43 new lesions that developed during the presymptomatic phase was located in a neurologically eloquent area, may be the reason why, for a relatively long period, the patient had no clinical manifestations of MS despite the marked MR findings of disease activity. PMID- 10472986 TI - St. Louis encephalitis and the substantia nigra: MR imaging evaluation. AB - Neuroimaging findings in cases of St. Louis encephalitis (StLE) have yet to be reported despite the relatively high frequency of this entity. An epidemic permitted the documentation of isolated hyperintensity of the substantia nigra on T2-weighted images in two patients with StLE. This distribution of MR imaging abnormality in cases of StLE mirrors the reports presented in the literature that implicate the substantia nigra as peculiarly susceptible to the StLE virus. Isolated lesions of the substantia nigra revealed by T2-weighted imaging should suggest the possibility of StLE. PMID- 10472985 TI - Prevalence and anatomic characteristics of infarct-like lesions on MR images of middle-aged adults: the atherosclerosis risk in communities study. AB - BACKGROUND AND PURPOSE: MR imaging has revealed putative evidence of subclinical cerebrovascular disease (CVD) as reflected by white matter signal changes and infarct-like lesions (ILLs). Nonetheless, the prevalence of this condition in the general population has been defined only to a limited extent. We herein report the prevalence and anatomic characteristics of ILLs seen on cranial MR images obtained as part of a population-based study of cardiovascular disease in middle aged adults. These results are contrasted to those of previous similar studies, particularly those of an elderly population in the Cardiovascular Health Study (CHS). METHODS: This Atherosclerosis Risk in Communities (ARIC) cohort consists of a probability sample of community-living persons who were 55 to 72 years old at the time of MR examination. MR imaging of 1890 participants was performed at two ARIC field centers, based on a common protocol. MR studies were evaluated by trained readers at the MR Reading Center using original digital data displayed on a high-resolution workstation. The measures of lesion size, anatomic location, and signal intensity were collected. The definition for an ILL was a non-mass, hyperintense region with an arterial vascular distribution on spin-density and T2 weighted images. RESULTS: Two hundred ninety participants had ILLs, for an overall prevalence of 15.3%. Eighty-two percent of participants with ILLs had lesions that were 3 mm or larger in maximal dimension, although 87% of these lesions were 20 mm or smaller in maximal dimension. The prevalence of ILLs increased with age, from 7.9% in the 55- to 59-year-old age group to 22.9% in the 65- to 72-year-old age group (P < .001). Lesion prevalence was greater in black (20.7%) than in white persons (10.2% [P < .0001]), but did not differ significantly between male and female participants. The basal ganglia and thalamic region was the most commonly affected anatomic site, accounting for 78.9% of the lesions. CONCLUSION: Considering that the prevalence of self reported stroke or transient ischemic attack in ARIC participants is 1.5%, these results suggest that there is significantly more subclinical than clinical CVD in the general population. Furthermore, the prevalence of this subclinical disease increases with age, and is greater in black persons. ILLs are dominated by "lacunae" in the basal ganglia and thalamus. These results are, in general, similar to those of a comparable study of elderly participants in the CHS, except for a 60% lower prevalence of ILLs in this younger population. PMID- 10472987 TI - Intracystic hemorrhage of the middle fossa arachnoid cyst and subdural hematoma caused by ruptured middle cerebral artery aneurysm. AB - We report a case of a cerebral aneurysm arising from the bifurcation of the left middle cerebral artery that ruptured into a left middle cranial fossa arachnoid cyst, associated with acute subdural hematoma. We discuss the relationships of aneurysm, arachnoid cyst, and subdural hematoma. PMID- 10472988 TI - Atypical Rathke's cleft cyst associated with ossification. AB - We report a case of symptomatic Rathke's cleft cyst with ossification. CT scans showed curvilinear calcification on the wall of the cyst. MR images revealed a cystic sellar lesion with a nodular solid mass extending to the floor of the third ventricle. This case shows that calcification of the suprasellar cyst does not always suggest craniopharyngioma. Rathke's cysts should be histologically differentiated from craniopharyngiomas because their treatments are different. PMID- 10472989 TI - Intraarterial thrombolysis for treatment of acute stroke: experience in 26 patients with long-term follow-up. AB - BACKGROUND AND PURPOSE: Since the approval of intravenous tissue plasminogen activator for acute ischemic stroke, great interest has been generated in cerebral fibrinolysis. Our purpose was to assess long-term outcome and hemorrhagic risk in patients with anterior circulation ischemic stroke treated with intraarterial urokinase. METHODS: Twenty-six patients were treated within 6 hours of ictus; of these, 21 were followed up for an average of 23 months. Angiographic reperfusion was classified according to thrombolysis in myocardial infarction (TIMI) grades. The Rankin Scale (RS) and the modified Barthel Index (mod BI) were used as outcome measures (good outcome: RS = 0-2, mod BI = 16-20; poor outcome: RS = 3-5, mod BI < or = 15). RESULTS: Ten of the 21 patients (average age, 48 years) had a good outcome; three (average age, 71 years) had a poor outcome; eight patients (average age, 78 years) died. Partial/complete (successful) recanalization was observed in 11 of 26 patients and minimal or no (unsuccessful) recanalization in 15. Recanalization favored a better outcome: nine of 21 had successful recanalization, with a good outcome in seven; 12 of 21 had unsuccessful reperfusion, with poor outcome/death in nine. Poor outcome was noted in five patients with internal carotid artery (ICA) bifurcation occlusions, four of whom had unsuccessful recanalization and poor outcome or death. Hemorrhage occurred in 10 of the 26 patients, with clinical deterioration in three. The average dose of urokinase was higher in the hemorrhage group, and mortality was higher in patients who hemorrhaged. CONCLUSION: Intraarterial thrombolysis is feasible in the setting of acute stroke. Successful reperfusion is associated with a better outcome, and the prevalence of hemorrhage does not exceed that which occurs in the natural history of embolic stroke. Poor outcome or death is associated with nonrecanalization, older age, hemorrhage, and ICA bifurcation occlusions. PMID- 10472990 TI - Definition of the ostium (neck) of an aneurysm revealed by intravascular sonography: an experimental study in canines. AB - BACKGROUND AND PURPOSE: The major factor influencing the effectiveness of Guglielmi detachable coils (GDCs) in the treatment of saccular aneurysms is the size of the aneurysm's ostium (neck). Current imaging techniques often do not allow accurate assessment of aneurysm neck morphology. The primary purpose of this study was to determine the feasibility of using intravascular sonography to provide this information. METHODS: Lateral and bifurcation aneurysms were created in each of six adult mongrel dogs by using a well-established surgical technique. Aneurysms were evaluated with digital subtraction angiography and intravascular sonography before (n = 12) and after (n = 6) treatment with GDCs. Angiography was performed using standard techniques. Sonography was performed using both a commercially available 2.6F 40-MHz catheter and a preproduction 0.014-inch 40-MHz imaging core wire housed in a Tracker catheter. Angiograms and sonograms were reviewed independently by two observers to assess the clarity and accuracy with which they depicted the size of each aneurysm's ostium. Posttreatment intravascular sonograms were evaluated for the extent to which they depicted the completeness of aneurysm obliteration. Two-dimensional reformatted images were made of the intravascular sonographic pullback sequences. RESULTS: In all instances, intravascular sonography provided clear definition of the aneurysm's neck (ostium) morphology as well as its relationship to the parent artery and adjacent branches, especially when 2D reformatted images were obtained. The position of coils in aneurysms was also clearly defined. CONCLUSION: Intravascular sonography is a novel technique for viewing the ostium (neck) of an aneurysm. It provides information not available with current angiographic methods. PMID- 10472991 TI - Saccular aneurysm formation in curved and bifurcating arteries. AB - BACKGROUND AND PURPOSE: Distinguishing whether forces resulting from the impingement of central blood flow streams at a curved arterial segment or at the apex of an intracranial bifurcation could be important for the understanding of aneurysm formation. Using finite element models, our purpose was to investigate the hemodynamics related to intracranial saccular aneurysm formation through computer simulations. METHODS: We present two-dimensional finite element models describing several distinct stages of aneurysm formation in both curved and bifurcating arteries. For each model, a description of the numeric solutions and results are presented. RESULTS: Our results suggest that the pressures and shear stresses that develop along the outer (lateral) wall of a curved artery and at the apex of an arterial bifurcation create a hemodynamic state that promotes saccular aneurysm formation. The impingement of the central stream results in greatly increased velocity/pressure gradients and high shear stresses at the apex compared with those in the proximal parent or distal daughter branches. The results also indicate that the maximal pressure generated at the apex of the arterial bifurcation ranges from two to three times the peak luminal pressure in the proximal parent artery. CONCLUSION: These data suggest that, in the absence of any underlying disease process, aneurysm development is a mechanically mediated event. These models offer a plausible hypothesis regarding the initiation, growth, and subsequent rupture of saccular intracranial aneurysms as they relate to the hemodynamics of intracranial arterial blood flow. PMID- 10472992 TI - Anomalous origin of the right vertebral artery: review of the literature and case report of right vertebral artery origin distal to the left subclavian artery. AB - We present the case of a 57-year-old patient who was admitted to the hospital for preoperative cerebral angiography because of an intraspinal mass at the level of C1 and C2. Angiographic examination revealed an abnormal origin of the right vertebral artery, which normally originates from the right subclavian artery. Thus, the right vertebral artery was the last branch of the supraaortic vessels. We also review herein the incidence of the various anomalous origins of the right vertebral artery in the literature and discuss their potential embryologic development and clinical significance. PMID- 10472993 TI - Embolization of cerebral arteriovenous malformations achieved with polyvinyl alcohol particles: angiographic reappearance and complications. AB - BACKGROUND AND PURPOSE: The appropriate choice of embolic materials with respect to the permanency of obliterated nidi after embolization and complications related to the procedure is essential for safe and effective embolization of cerebral arteriovenous malformations (AVMs). Our purpose was to ascertain the recanalization and complication rates after AVM treatment with polyvinyl alcohol (PVA) particles. METHODS: Between 1988 and 1994, 36 AVMs were embolized with PVA particles at our institution. Follow-up angiographic findings and occurrence of complications during the embolization procedures were analyzed retrospectively. RESULTS: Complete obliteration of the nidus immediately after embolization was achieved in five patients, and 80% to 99% obliteration was attained in 12 patients. Fifty-one follow-up angiographic examinations were performed 1 week to 60 months (mean, 7 months) after embolization in 31 patients. An increase in nidal size was seen on 15 follow-up angiograms (29%) and a decrease was seen in seven (14%). In 28 of the 51 angiograms obtained more than 1 month after follow up (mean, 13 months), 12 (43%) showed AVM enlargement. In four (80%) of five cases of complete obliteration, nidi reappeared on follow-up angiograms. Hemorrhagic complications occurred in three cases and ischemic ones in seven. One patient (3%) died and five (14%) suffered persistent neurologic deficits. CONCLUSION: Embolization with PVA particles can produce significant volume reduction in AVM nidal size, but recanalization is a distinct possibility. PMID- 10472994 TI - Intraventricular contrast medium leakage during ethanol embolization of an arteriovenous malformation. AB - We report the unusual phenomenon of abrupt intraventricular contrast medium leakage from the choroid plexus occurring during ethanol embolization of a periventricular arteriovenous malformation. There was no evidence of any associated intraventricular hemorrhage to suggest that leakage arose from a vessel perforation, as was first suspected. Intraventricular contrast medium leakage has been reported previously in the setting of ependymitis, and it is likely that similar pathogenetic mechanisms apply in this case. To our knowledge, this is the first reported case of intraventricular contrast medium leakage occurring during an embolization procedure. PMID- 10472995 TI - Three-dimensional multivoxel proton MR spectroscopy of the brain in children with neurofibromatosis type 1. AB - BACKGROUND AND PURPOSE: Neurofibromatosis type 1 (NF1), the most common autosomal dominant genetic disorder, frequently manifests as focal areas of signal intensity (FASI) on T2-weighted MR images. The purpose of our study was to investigate whether tumor(s), focal areas of signal intensity (FASI), and normal brain can be differentiated by using 3D multivoxel localized proton MR spectroscopy in children with neurofibromatosis type 1 (NF1) disorder. METHODS: Five children with NF1 and two healthy control subjects, all in the 3- to 11-year old age group, were studied with a new 3D proton MR spectroscopy technique: a hybrid of 1D fourth-order transverse Hadamard spectroscopic imaging and 2D chemical shift imaging. A 3D volume-of-interest (VOI) was image-guided onto the site of the abnormality and identified on three orthogonal images. Proton MR spectroscopy partitioned the VOI into 6 x 6 x 4 (or 8 x 8 x 4) voxels, 1.5 (or 1.0) cm3 each. RESULTS: Simultaneous coverage of the entire VOI yielded good spectral signal-to-noise ratio from 136 (or 256) voxels in 27 minutes. Proton MR spectroscopy indicated that FASI a) are characterized by significantly elevated choline (Cho), reduced creatine (Cr), 2>Cho: Cr>1.3, and near normal N acetylaspartate (NAA) levels; b) are different from tumors that exhibit Cho:Cr>2 and no NAA; c) have no intrinsic lipid or lactate signal(s); and d) correlate in spatial extent but are more extensive than indicated by MR imaging. CONCLUSION: Three-dimensional multivoxel proton MR spectroscopy reveals distinct metabolic features that differentiate normal, FASI, and tumor regions in the pediatric brain. PMID- 10472996 TI - Reproducibility and accuracy of MR imaging of the brain after severe birth asphyxia. AB - BACKGROUND AND PURPOSE: MR imaging of the brain can be used to detect cerebral damage after suspected hypoxic-ischemic injury. This study examines the reproducibility and accuracy of MR imaging soon after severe birth asphyxia. METHODS: During a 48-month period, full-term newborn neonates, who died within the first week as a result of severe hypoxic ischemic encephalopathy, were included in the study if they had undergone early (<5 days old) MR imaging and postmortem neuropathologic studies. Two trained observers assessed reproducibility by examining multiple brain regions independently with current criteria and then defining and applying improved criteria. Accuracy of MR findings was tested by comparing the brain regions about which the two imaging raters agreed to those regions about which the two pathologists agreed. RESULTS: Eight neonates, with a median gestational age of 40 weeks (range, 38-40 weeks) and who suffered severe birth asphyxia, were included in the study. In the reproducibility study, MR imaging agreement was moderate when current criteria were used (k = .44). Using the improved criteria, agreement increased considerably (k = .62). Much of this improvement was due to limiting the analyses to the posterior limb of the internal capsule, thalamus, parietal cortex, hippocampus, and medulla. The posterior limb of the internal capsule was the most reliable region analyzed. MR imaging agreement was similar to that achieved by two experienced pathologists reviewing the histologic sections (k = .66). In the accuracy study, MR imaging abnormality was predictive of pathologic abnormality with a sensitivity of .79 and a positive predictive value of 1.0. The predictive value of a single MR imaging abnormality was .79 (95% confidence interval, .61 .96). CONCLUSION: Criteria that provide substantial reproducibility and accuracy for the interpretation of MR imaging findings very early after birth asphyxia can be derived. PMID- 10472997 TI - Relationship between MR imaging and histopathologic findings of the brain in extremely sick preterm infants. AB - BACKGROUND AND PURPOSE: MR imaging can now be used safely in extremely preterm infants. The aim of this study was to compare the MR imaging appearance of the immature brain with neuropathologic findings at postmortem examination. METHODS: Seven extremely sick preterm infants, born at a median of 24 weeks' gestation, were studied using T1- and T2-weighted MR sequences. Infants died at a median of 3 days after initial MR imaging, and postmortem examinations were carried out. RESULTS: The cortex and germinal matrix were seen as areas of low signal intensity on T2-weighted images, which corresponded to their highly cellular histologic appearance. The periventricular and subcortical layers of white matter had a high signal intensity, corresponding to high fiber and relatively low cellular density; the intermediate layer of low signal intensity corresponded to a dense band of migrating cells. Regions of acute hemorrhage were seen as low signal intensity and regions of infarction as high signal intensity on T2 weighted images. One infant with mild periventricular leukomalacia had some low signal intensity on T1-weighted images, but no focal changes on T2-weighted images. Regions of neuronal mineralization, seen in association with infarction and capillary proliferation, within the basal ganglia and thalami were characterized by very low signal intensity on T2-weighted images and by very high signal intensity on T1-weighted images. There were no imaging abnormalities detected in regions with more subtle histologic abnormalities, such as increased glial or apoptotic cells. CONCLUSION: MR imaging can be used to observe normal developing brain anatomy in extremely premature infants; it can detect areas of hemorrhage and infarction within the developing brain, but conventional MR imaging may not detect more subtle histologic abnormalities. PMID- 10472998 TI - Sonographic nomogram of the leptomeninges (pia-glial plate) and its usefulness for evaluating bacterial meningitis in infants. AB - BACKGROUND AND PURPOSE: To our knowledge, the upper limits of the thickness of normal meninges on neurosonograms are not known. We therefore established a nomogram for sonographic measurements of the leptomeninges (pia-glial plate) and assessed its usefulness in neurosonographic examinations of children with bacterial meningitis. METHODS: The pia mater-cortical glia limitans complex on the surface of the brain and in the sulcus of a frontal gyrus was measured on neurosonograms in 100 infants without meningeal disease in order to establish a nomogram of the thickness of this pia-glial plate, referred to as the leptomeninx. Effects of prematurity, age, sex, and single-layer (surface) versus double-layer (sulcus) measurements were analyzed. Meningeal thicknesses derived from a retrospective analysis of the neurosonograms of 33 patients with purulent meningitis and a prospective study of 22 patients with bacterial meningitis were compared with the nomograms. Clinical outcomes of children with meningeal thickening were compared with those of affected children with normal meninges. RESULTS: The distribution of sulci measurements was significantly asymmetrical around the mean. Statistical data showed no influence of prematurity and sex, but showed surface measurements to be more consistent than sulcal measurements. Older chronological age was related to slightly larger sulci, but did not influence the surface measurements. In children with bacterial meningitis, the surface meninges were less frequently thickened than were the sulci. Sulcal enlargement occurred often in combination with echogenic deposits in the sub-arachnoid space. CONCLUSION: Leptomeninges are best measured on the surface of a gyrus rather than in a sulcus, as the normal thickness of the sulci shows much more variability. Clinical outcome of bacterial meningitis cannot be predicted by presence or absence of meningeal thickening as the only sonographic abnormality. PMID- 10472999 TI - Acute spontaneous spinal epidural hematomas. AB - BACKGROUND AND PURPOSE: Although previous reports have characterized MR imaging features of spinal epidural hematomas (EDH), few cases have been reported during the acute or hyperacute phase within the first 48 hours. Our goal in this investigation was to correlate the MR imaging features of acute (< or =48 hours) spontaneous EDH with clinical management and outcome. METHODS: Eight patients with acute spontaneous EDH (five men and three women; age range, 31-81 years) underwent MR imaging at 1.5 T (T1-weighted, n = 8; T1-weighted after the administration of 0.1 mmol/kg contrast material, n = 6; T2-weighted, n = 8; and T2-weighted, n = 4). The interval from symptom onset to hospital admission ranged from immediate to 5 days. Two neuroradiologists reviewed the MR images for signal characteristics, contrast enhancement, and cord compression. Treatment and clinical outcome were correlated with the imaging findings. RESULTS: The EDH were located in the cervical (n = 3), cervicothoracic (n = 2), thoracolumbar (n = 2), and lumbar (n = 1) regions. On T1-weighted images, the signal intensity of the EDH was isointense to spinal cord in five cases, hyperintense in two cases, and hypointense in one case and did not correlate with time to imaging. Isointensity on T1-weighted images persisted for 5 days in one case. On T2-weighted images, all EDHs were hyperintense with focal, heterogeneous hypointensity. Cord compression was severe in six patients, moderate in one patient, and minimal in one patient. Four cases were treated conservatively with complete resolution or improvement of symptoms within 1 to 3 weeks. CONCLUSION: MR imaging findings were useful in establishing the diagnosis of EDH but did not influence management or predict outcome in this series. Heterogeneous hyperintensity to cord with focal hypointensity on T2-weighted images should suggest the diagnosis of acute spinal EDH. Severity of neurologic impairment had the greatest impact on management and outcome. Nonoperative treatment may be successful in cases with minimal neurologic deficits, despite cord compression revealed by MR imaging. PMID- 10473000 TI - Intraspinal extradural cysts communicating with adjacent herniated disks: imaging characteristics and possible pathogenesis. AB - We report two cases of intraspinal extradural cysts communicating with an adjacent herniated disk that we term "disk cysts." These cysts were well defined and homogeneous, and were present in the ventrolateral extradural space adjacent to a lumbar herniated disk. They had rim enhancement on contrast-enhanced MR images, and communication with a herniated disk was revealed by diskography. PMID- 10473001 TI - The status of carotid stenting. PMID- 10473002 TI - Gradient- and spin-echo MR imaging of the brain. PMID- 10473003 TI - The evolutionary and embryologic basis for the development and anatomy of the cavum veli interpositi. PMID- 10473004 TI - Gender differences in drug treatment careers among clients in the national Drug Abuse Treatment Outcome Study. AB - Gender differences in the factors associated with having a history of drug treatment were examined among 7,652 individuals admitted into the Drug Abuse Treatment Outcome Study (DATOS), a national multisite prospective study. Bivariate relationships were examined between a history of prior drug treatment and variables measuring demographic and background characteristics, addiction career, treatment career parameters, family and social relationships, criminal justice involvement, and mental health status. Stepwise discriminant function analyses (DFAs) were conducted separately for men and women to determine both the common and unique characteristics associated with a history of prior drug treatment. More severe drug use history and criminal behavior were related to prior treatment history for both men and women. Prior drug treatment among men was associated with factors related to family opposition to drug use and support for treatment, whereas for women prior drug treatment was associated with antisocial personality disorder and self-initiation into treatment. Moreover, treatment initiation among men appears to be facilitated by social institutions, such as employment, the criminal justice system, and one's family. In contrast, treatment reentry among women was associated with referral by a social worker, suggesting that contact with family service agencies can facilitate women's treatment entry. The findings suggest that different strategies for increasing treatment utilization may be appropriate for men and women. PMID- 10473005 TI - Positive effects on life skills motivation and self-efficacy: node-link maps in a modified therapeutic community. AB - In recent years, one response to drug abuse problems has been to provide treatment in prison and probation settings. Results are promising, although the need for improving mandated treatment has been expressed. The Cognitive Enhancements for Treatment of Probationers (CETOP) project is investigating cognitive enhancements in a modified therapeutic community (TC) setting. One enhancement is node-link mapping, a visual graphing strategy. Map "nodes" contain ideas, actions, and feelings; these are connected by links that illustrate meaningful relationships. Current findings indicate increases in group participation, ratings of session depth, and positive ratings of coresidents, counselors, and security staff. The present study extends this research by comparing mapping-enhanced counseling to standard counseling on self-efficacy and motivation for basic psychosocial skills (e.g., emotional control). Probationers (n = 381) in a 16-week residential program were assigned randomly to conditions in 12 TCs (n = 30-35). Motivation and self-efficacy were assessed by two self report questionnaires, one midway and one at the end of treatment. A five-factor solution was produced from each questionnaire. With community as the unit of analysis, means of enhanced counseling were higher in all cases. Wilcoxen tests indicated four significant differences at midterm for mapping on motivation and self-efficacy of communication and emotional control (p < .05). At the end of treatment, mapping was higher for motivation of cognition and emotional control and self-efficacy was higher for communication. Discussing Bandura's model, these findings provide evidence that maps may enhance psychosocial skills, which have been associated with maintaining recovery. PMID- 10473006 TI - Violence in the lives of pregnant teenage women: associations with multiple substance use. AB - Using data from a consecutive sample of approximately 700 prenatal patients aged 12 to 19, the extent of violence and substance use in the lives of these women was examined, as were associations between violence and substance use. The findings show that 29% of the study participants had been victims of violence, with 15% experiencing physical violence only, 5% sexual violence only, and 9% both physical and sexual violence. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) from logistic regression analyses showed that, after controlling for confounding factors, victims were more likely than nonvictims to use cigarettes, alcohol, and illicit drugs, with victims of both physical and sexual violence being the most likely to use each type of substance. Ordinal logistic regression analysis found that, compared to nonvictims, violence victims were more likely to have more severe patterns of substance use (use multiple types of substances), with victims of both physical and sexual violence being the most likely to be multiple substance users. These findings underline the importance of both violence and substance use as health concerns among our youth. PMID- 10473007 TI - Attention-deficit/hyperactivity disorder and substance use: symptom pattern and drug choice. AB - While there has been much recent interest in the relationship between attention deficit/hyperactivity disorder (ADHD) and substance use disorders (SUDs), little has been reported about ADHD diagnostic subtypes, persistence of symptoms from childhood into adulthood, and substance of choice in individuals with substance use disorders (SUD+) and comorbid ADHD. To examine the prevalence and subtypes of ADHD in a group of SUD+ individuals, 136 inpatients with an SUD diagnosis (cocaine vs. alcohol vs. cocaine/alcohol) were administered a structured interview for ADHD. Of the SUD+ individuals, 32% met criteria for ADHD, and 35% of those with a childhood diagnosis of ADHD continued to have clinically significant symptoms into adulthood. There were no significant differences in the percentage of ADHD between the SUD+ groups divided by drug choice. Of ADHD subtypes, subjects with combined and inattentive types were significantly more likely to have symptoms continue into adulthood (p < or = .05) than the hyperactive/impulsive subtype. Patients with cocaine use were more likely to have ADHD in childhood only when compared to the alcohol or cocaine-alcohol groups. The findings of this study indicate that ADHD is prevalent in treatment-seeking substance users without difference in prevalence or subtype by drug choice. PMID- 10473008 TI - Prevalence of childhood trauma in a community sample of substance-abusing women. AB - The purpose of this study was to estimate the prevalence of positive trauma histories in a community sample of intravenous drug using (IVDU) women for five subsets of childhood trauma (emotional abuse or neglect, physical abuse or neglect, and sexual abuse) and to compare demographic variables between the abused versus nonabused groups. Of the 181 women who completed the Childhood Trauma Questionnaire (CTQ), 109 (60.2%) were sexually abused, 100 (55.2%) were physically abused, 83 (45.9%) were emotionally abused, 151 (83.4%) were emotionally neglected, and 108 (59.7%) were physically neglected. There were no statistically significant findings for age, ethnicity, and educational level. Those subjects that were physically neglected were more likely not to be in a current relationship compared to those subjects that were not abused (p = .036). The findings suggest that the prevalence of all five childhood traumas was higher than what has been reported in the general population, and that physical neglect of individuals may predict lack of current significant relationships. PMID- 10473009 TI - Drug network characteristics as a predictor of cessation of drug use among adult injection drug users: a prospective study. AB - Few studies have examined recovery from opiate and cocaine dependence without treatment, referred to as "natural recovery," "spontaneous recovery," and "spontaneous remission." The present study examined the relationship between network characteristics and cessation of heroin, cocaine, and crack use in a sample of underclass inner-city injection drug users in Baltimore, Maryland. Participants were enrolled in an experimental human immunodeficiency virus (HIV) preventive intervention. Between the baseline and follow-up interviews, which averaged 5.2 months, 24 (7%) of 335 participants reported ceasing to use heroin, cocaine, and crack. Individuals who had reported cessation of drug use at follow up had reported at baseline a smaller proportion of their network members with whom they used drugs (p < .02). Using multiple logistic regression analyses and adjusting for baseline drug use, enrollment in drug treatment, and demographic and background variables, cessation of drug use was associated with a lower proportion of personal network members in one's drug network (odds ratio [OR] = 25.4, p < .05). The data from this study suggest that network members have potential for social influence on the cessation of drug use. PMID- 10473010 TI - Methamphetamine use and sexual and injection risk behaviors among out-of treatment injection drug users. AB - Our primary objective was to examine the relationship between methamphetamine use and sexual risk-taking behaviors--number of sexual partners, frequency of sexual behaviors with regular and casual partners, trading money or drugs for sex, and condom use--among male and female out-of-treatment injection drug users (OTIDUs). As a risk group for human immunodeficiency virus (HIV) transmission, we also investigated injection behaviors by methamphetamine use. Data were collected from 1392 OTIDUs within the California counties of Fresno, Sacramento, and San Diego. Excluded from this cross-sectional survey were male OTIDUs engaging in sex with only or mostly men since 1978. In bivariate analyses, we found that male OTIDUs with a history of methamphetamine use had more sex partners and participated in more acts of anal insertive intercourse with casual partners and vaginal intercourse with regular and casual partners than male OTIDUs never using methamphetamines. In addition, a greater percentage of male OTIDUs using methamphetamines reported trading sex for money or drugs. Methamphetamine-using female OTIDUs participated in more acts of vaginal intercourse with regular male sex partners than female OTIDUs never using methamphetamines. By multivariate logistic regression, we found methamphetamine use related to consistent condom use among male OTIDUs and among male sex partners of female OTIDUs. Discriminant function analyses revealed that sexual risk taking could be differentiated by methamphetamine use among male OTIDUs. Methamphetamine use also correlated with using shared needles or syringes among male and female OTIDUs and was related to not always disinfecting used needles or syringes with bleach. Our findings suggest that methamphetamines may contribute to heterosexual HIV transmission. PMID- 10473012 TI - Some sex-neutral and sex-specific factors related to employment among substance abuse clients. AB - Workfare programs, designed to implement the 1996 federal Personal Responsibility and Work Reconciliation Act, raise anew the issue of the employability of substance abusers. The present study explores this issue by examining factors potentially related to the employment status at intake of 1274 men and 804 women admitted for substance abuse treatment to the Center for Comprehensive Health Practice in New York City from July 1995 through December 1997. It was found that men were more likely to be employed than women. For both sexes, employment was positively related to education and stable housing. For men, employment was negatively related to having a mobility impairment; for women, it was negatively related to their number of children and being pregnant. For each sex, variables significantly associated with employment were used to construct a "work barriers" scale. For both sexes, employment was negatively related to number of work barriers. Women generally faced more barriers than men, accounting for their lower employment rate. Public policy and research implications of the findings are discussed. PMID- 10473011 TI - Modified therapeutic community for homeless mentally ill chemical abusers: emerging subtypes. AB - This paper is one of a series reporting on a clinical field trial evaluating the efficacy of the modified therapeutic community (TC) approach for the treatment of homeless mentally ill chemical abusers (MICAs). The social and psychological characteristics of the treatment sample were described in an earlier paper; the purpose of the present report was to categorize subtypes of homeless MICA clients to predict with greater accuracy their treatability in modified TCs. An index that consistently correlated with treatment-relevant variables was identified for each of three dimensions; Homelessness (residential instability), Mental Illness (current severity), and Substance Abuse (current substance abuse/dependence diagnosis). These indices yielded distributions that captured the variability in the sample with respect to a number of variables, including drug use, criminality, human immunodeficiency virus (HIV) risk (sexual behavior), psychological status, and motivation. Bivariate and multivariate analyses showed that the indices were not strongly related to demographic variables such as race/ethnicity, age, or gender, but were significantly associated with baseline drug use, criminal activity, HIV risk (sexual behavior), psychological symptoms, and motivation and readiness. These findings indicate that, even among those admitted to residential treatment for substance abuse, homeless MICA clients are not homogeneous; rather, subgroup differences emerge among the indices of homelessness, mental illness, and substance abuse. The efficacy of treatment in modified TCs for these subgroups will be assessed in subsequent papers examining the relationships among the three indices, client retention, and outcomes during and subsequent to residential treatment. PMID- 10473013 TI - The effect of alcohol abuse on the health status of older adults seen in the emergency department. AB - OBJECTIVE: To examine the effect of alcohol abuse on the subsequent health status of elderly patients seen in an emergency department (ED). PATIENTS AND METHODS: A sample of 966 patients aged 65 or older who presented to one urban academic ED between the hours of 8 A.M. and 12 midnight was followed for 1 year. A personal interview was administered during the ED visit. Current problem drinkers had a score of 1 or greater on the CAGE questionnaire at ED presentation and drank within the prior 6 months; former problem drinkers had a score of 1 or greater on the CAGE questionnaire at ED presentation and a last drink more than 6 months previously. We used 13 items from the Medical Outcomes Study short form adapted to the ED setting and 6 items from the Index of Activities of Daily Living (ADL) to measure health status. RESULTS: In multivariate models for repeated-measures controlling for potential confounding factors, current problem drinkers had worse overall health (parameter estimate beta -3.6; 95% CI -7.1 to -0.04), and former problem drinkers had worse mental health (beta -3.6; CI -6.9 to -0.24) on follow up. We could find no effect of problem drinking on physical health or social function. CONCLUSIONS: Current problem drinking is associated with worse self perceived health among elderly patients in the year following presentation to an ED. The magnitude of decline in health perception may approximate the effect of having back pain, sciatica, or other musculoskeletal complaints. Elderly former problem drinkers suffer from more severe mental health problems over that same period. PMID- 10473014 TI - Volunteer assistance in the treatment of chronic alcoholism. AB - Chronic alcoholics often have great difficulty in adjusting their lifestyle to accommodate the goals agreed upon in treatment. A 16-week behavioral self management program, which emphasized creating lifestyle changes in the community, was offered to 193 clients. To assist in the process of treatment delivery, half of the clients were offered the support of trained community volunteers during the treatment program. To give volunteers a time period in which to to meet and start working with their clients, only those clients who completed at least the first 4 weeks of the 16-week program (N = 106) were included in the evaluation. Independent follow-up was conducted over a 12-month period. A significant reduction in alcohol consumption was found for clients of both the volunteer support (VS; N = 52) and the office-based (OB; N = 54) groups; this reduction was maintained over the 12 months of review, with no major differences found between groups. Volunteers rated high on particular characteristics assisted the client more effectively during treatment than those rated low. The variability in hours spent between volunteer-client pairs appeared to mask any main effects, and also the length of the contact period with the volunteer was too short. It is recommended that the use of volunteers be evaluated further by extending the client contact time into the follow-up period. PMID- 10473015 TI - Detection of illicit opioid and cocaine use in methadone maintenance treatment. AB - Urine toxicology is the gold standard for estimating the prevalence of illicit drug use in methadone maintenance treatment (MMT). The frequency of urine testing may be crucial for establishing accurate use rates. Infrequent testing may lead programs to undercount active drug users and to target interventions too narrowly. This study compared results from frequent testing (twice per week) versus less frequent testing of 166 patients at four MMT programs. As part of a research study, all patients were tested by research staff for opioid and cocaine use twice per week on a fixed schedule for 10 weeks. During the same period, the four MMT programs tested the patients according to their standard protocols, approximately weekly (one program) or every 3-4 weeks (three programs). The research tests identified approximately 50% more illicit opioid users and 70% more cocaine users than the less frequent program tests. Patients who were drug positive according to the research tests but drug negative according to the program tests tended to be infrequent users. The data suggest that standard urine testing practices in MMT programs may result in underestimates of the prevalence of opioid and cocaine use. More frequent testing, even for time-limited periods, should produce more accurate depictions of drug use prevalence and help indicate the direction of interventions. PMID- 10473016 TI - Beta-blockers in heart failure. The 'new wave' of clinical trials. AB - There is now considerable clinical trial data to support the use of beta-blockers in patients with congestive heart failure (CHF) due to systolic left ventricular dysfunction. A substantial database has accumulated over the last 20 years supporting the benefits of these agents on ventricular function and clinical status. In addition, morbidity and mortality benefits have been suggested, specifically with the non-selective vasodilating agent, carvedilol. More recently, a "new wave" of clinical trials have been conducted to definitively determine the mortality benefits of beta-blockers in patients with mild to moderate CHF as well as addressing other important clinical questions. These questions include whether the beneficial effects of carvedilol on survival can be reproduced by other agents in prospective, adequately powered studies; whether the benefits of carvedilol in systolic heart failure are due to its beta-blocking properties alone or to a combination of the beta-blocking and ancillary effects of the drug; whether beta-blockers are of benefit in patients with severe New York Heart Association (NYHA) Class IIIB-IV CHF; and, whether beta-blockers are of benefit (additional to ACE inhibitors) in patients with evidence of systolic ventricular dysfunction when commenced in the immediate post-myocardial infarction period. Major studies are currently being undertaken to address the above questions. Most are still underway but 3 studies have recently reported their results: the second Cardiac Insufficiency Bisoprolol Study (CIBIS II), the Research in Left Ventricular Dysfunction Study (RESOLVD), and the Metoprolol CR/XL Randomised Intervention Trial in Heart Failure (MERIT-HF) study. These studies have demonstrated that blockade with beta1-selective, non-vasodilating agents (i.e. bisoprolol and metoprolol) improve survival in patients with CHE Comparison of relative risk reduction in these recent studies with the earlier carvedilol studies raises mechanistic questions, specifically whether non selectivity, vasodilation and other ancillary properties of carvedilol are critical to its benefit in CHF patients. This question is currently being addressed in the Carvedilol and Metoprolol European Trial (COMET), comparing metoprolol with carvedilol. The beneficial effects of beta-blockers on mortality in patients with mild to moderate CHF have also had major implications in ongoing studies of other agents in this condition. Open-label prescribing of beta blockers is increasing in these studies and this is having an impact on event rates and thus required duration of administration of study drug. Furthermore, it would now appear unethical to deprive suitable NYHA Class II-III CHF patients of beta-blockers as part of the design of such studies. In conclusion, beta-blockers have now become the most extensively studied class of agents in the treatment of CHF, with a database of over 6000 patients in placebo-controlled studies, and ongoing clinical and mechanistic studies. Despite this, further questions remain regarding the use of these agents in CHF, including their role in the extreme elderly, in patients with diabetes mellitus and in patients with renal impairment. PMID- 10473018 TI - Comprehensive pharmacology and clinical efficacy of aromatase inhibitors. AB - The goal of hormone therapy is to deprive breast tumours of estrogens, since estrogens have been implicated in the development or progression of tumours. This can be accomplished by the use of antiestrogens that block estrogen action or by inhibiting aromatase, the enzyme that catalyses the final and rate-limiting step in estrogen biosynthesis. A number of steroidal and nonsteroidal compounds have been developed as aromatase inhibitors. This review highlights the valuable role that a few of these aromatase inhibitors have played, and continue to play, in the treatment of breast cancer. Following background information regarding the biochemistry of aromatase, the rationale for its inhibition, and an outline of the test systems for evaluating and characterising aromatase inhibitors, the discussion focuses on the new generation of aromatase inhibitors that are in clinical trials or clinically available. Specifically, it discusses the pharmacology and clinical efficacy of formestane, exemestane, rogletimide, fadrozole, vorozole, anastrozole and letrozole. The role of these agents as the optimal second-line agents (after tamoxifen) for the treatment of advanced breast cancer has been established; their prospects in other clinical settings and as potential breast cancer chemopreventives are warranted but are yet to be fully determined. PMID- 10473021 TI - Perindopril 2mg/indapamide 0.625mg. Fixed low-dose combination. AB - Low-dose drug combinations have been proposed in International Guidelines for use in patients with hypertension. The fixed low-dose combination of perindopril 2mg with indapamide 0.625mg combines an angiotensin converting enzyme (ACE) inhibitor with a non-thiazide diuretic. Coadministration of perindopril and indapamide did not have any clinically significant effects on the pharmacokinetic profile of either agent in healthy volunteers. In experimental models of hypertension, perindopril/indapamide restored endothelial function, improved microvascular density, reduced left ventricular and aortic hypertrophy, and reversed renal end organ damage. Once daily oral perindopril 2mg/indapamide 0.625mg normalised blood pressure (BP) in 83.6% of elderly patients with essential hypertension (diastolic BP was reduced to < or =90mm Hg) and 81.7% of those with isolated systolic hypertension (systolic BP was reduced to <160mm Hg) after approximately 1 year of treatment. BP normalisation was sustained in 79.8% of patients throughout the study. Fixed low-dose perindopril/indapamide had a tolerability profile similar to that of placebo in clinical trials; most adverse events were of mild to moderate severity. Coadministration of the 2 agents reduced the incidence of hypokalaemia seen with indapamide alone. PMID- 10473020 TI - Management of onychomycoses. AB - Onychomycoses, infections of the nail caused by fungi, are amongst the most common illnesses. Because of the high incidence of these infections and problems involved in their therapy, they have received much attention, particularly as concerns a better characterisation of the causative micro-organisms. Onychomycosis caused by dermatophytes (tinea unguium) is most common and is found more frequently on the feet than on the hands. The clinical presentation of onychomycosis is at best indicative of fungal infection, and the growth of a credible pathogen is an indispensable prerequisite for definite diagnosis. The clinical appearance is variable. Four major types of manifestation have been characterised, depending on localisation and spread. New antifungal agents for systemic or topical application based on novel active substances or vehicles are available, and cure is feasible for the majority of cases. Therapy can and should be individualised, depending on the characteristics of the particular case. Currently, continuous or intermittent oral treatment with itraconazole or terbinafine exhibit a particularly favourable risk : benefit ratio. Fluconazole might become an alternative in the near future. With respect to topical treatment, ciclopirox or amorolfine lacquer and the bifonazole/urea combination deserve particular interest. However, cure cannot be expected for every case. PMID- 10473017 TI - Adenosine triphosphate: established and potential clinical applications. AB - Adenosine 5'-triphosphate (ATP) is a purine nucleotide found in every cell of the human body. In addition to its well established role in cellular metabolism, extracellular ATP and its breakdown product adenosine, exert pronounced effects in a variety of biological processes including neurotransmission, muscle contraction, cardiac function, platelet function, vasodilatation and liver glycogen metabolism. These effects are mediated by both P1 and P2 receptors. A cascade of ectonucleotidases plays a role in the effective regulation of these processes and may also have a protective function by keeping extracellular ATP and adenosine levels within physiological limits. In recent years several clinical applications of ATP and adenosine have been reported. In anaesthesia, low dose adenosine reduced neuropathic pain, hyperalgesia and ischaemic pain to a similar degree as morphine or ketamine. Postoperative opioid use was reduced. During surgery, ATP and adenosine have been used to induce hypotension. In patients with haemorrhagic shock, increased survival was observed after ATP treatment. In cardiology, ATP has been shown to be a well tolerated and effective pulmonary vasodilator in patients with pulmonary hypertension. Bolus injections of ATP and adenosine are useful in the diagnosis and treatment of paroxysmal supraventricular tachycardias. Adenosine also allowed highly accurate diagnosis of coronary artery disease. In pulmonology, nucleotides in combination with a sodium channel blocker improved mucociliary clearance from the airways to near normal in patients with cystic fibrosis. In oncology, there are indications that ATP may inhibit weight loss and tumour growth in patients with advanced lung cancer. There are also indications of potentiating effects of cytostatics and protective effects against radiation tissue damage. Further controlled clinical trials are warranted to determine the full beneficial potential of ATP, adenosine and uridine 5'-triphosphate. PMID- 10473019 TI - Diagnosis and treatment of patients with testicular germ cell cancer. AB - Testicular germ cell tumours are a highly curable malignancy even in the presence of metastases, with an overall survival rate of approximately 90 to 95% when all stages are considered. Current therapeutic strategies aim at risk-adapted therapy, trying to maintain favourable overall results while reducing treatment related toxicity, particularly in non-advanced disease. In stage I disease, unilateral inguinal orchiectomy represents the standard diagnostic and therapeutic approach. For patients with clinical stage I seminoma, prophylactic para-aortic radiotherapy with 26Gy is commonly employed. In patients with nonseminomatous germ cell tumours (NSGCT) at clinical stage I, the 3 options are: (i) retroperitoneal lymphadenectomy; (ii) a wait-and-see strategy; or (iii) 2 cycles of adjuvant chemotherapy. The individual risk profile for tumour recurrence, mainly based on histopathological criteria such as vascular tumour invasion, should guide treatment decisions in this group of patients. Radiotherapy is still the standard approach in clinical stage IIA/B seminoma, whereas in patients with a low tumour burden of NSGCT, retroperitoneal lymphadenectomy is frequently used followed by surveillance or adjuvant chemotherapy. Primary chemotherapy in these stages of disease may be at least equally successful. Major progress has also been achieved in the treatment of NSGCT patients with metastatic disease greater than clinical stage IIB, for whom primary chemotherapy represents the standard approach. After cisplatin-based combination chemotherapy, between 70 and 90% of patients will achieve a durable remission. In patients with 'good risk' metastatic disease, 3 cycles of cisplatin, etoposide and bleomycin (PEB) remain the standard treatment, despite several randomised trials trying to avoid the lung-toxic bleomycin or substituting cisplatin by carboplatin. In patients with 'intermediate' and 'poor prognosis' disease, 4 cycles of PEB given at 3-week intervals are considered routine treatment. The role of high dose chemotherapy with peripheral autologous blood stem cell transplantation is currently being investigated for patients presenting initially with advanced (poor prognosis) metastatic disease and for patients with relapse after previous chemotherapy, in whom conventional-dose salvage regimens will only result in 20% long-term survival. Because of the large group of patients with metastatic disease being cured, the possible long-term adverse effects of treatment have become important. Only a slightly elevated risk for therapy-related secondary malignancies has been identified. Long-term adverse effects associated with cisplatin may affect a larger proportion of patients. Further research should focus on reducing the risk of chemotherapy-related chronic toxicity. PMID- 10473022 TI - Palivizumab. AB - The humanised monoclonal antibody palivizumab has been developed for prevention of serious lower respiratory tract disease caused by respiratory syncytial virus (RSV) in infants at high risk; RSV is the most common cause of lower respiratory tract infections in infants. Palivizumab specifically inhibits an epitope at the A antigenic site of the F protein of RSV subtypes A and B. RSV replication was inhibited in nasal and tracheal aspirates from infants receiving palivizumab 15 mg/kg. Mean 30-day trough serum concentrations of palivizumab were consistently about 70 mg/L in infants receiving repeated intramuscular or intravenous palivizumab 15 mg/kg. This is above the target serum concentration of 40 mg/L estimated to reduce pulmonary RSV replication by >99% in animal studies. In a large multicentre trial in 1502 infants at high risk of RSV infection, intramuscular palivizumab 15 mg/kg more than halved the incidence of RSV attributable hospitalisation to 4.8% compared with 10.6% in placebo recipients. In the same group of high-risk infants, palivizumab significantly decreased total days in hospital attributable to RSV infection, days with increased supplemental oxygen requirement, days with moderate to severe lower respiratory tract infections and the incidence of admissions to intensive care. It had no effect on the incidence or total number of days of ventilation. Palivizumab was well tolerated during clinical trials in infants at risk of RSV infection. The incidence of adverse events was similar in placebo (10%) and palivizumab (11%) groups. Fever, irritability and injection site reaction were the most commonly reported adverse events. PMID- 10473023 TI - Intranasal metoclopramide. AB - Intranasal metoclopramide is a new formulation of an established and effective antiemetic drug. Absorption after intranasal administration was lower than after oral or intravenous administration; otherwise the pharmacodynamic and pharmacokinetic profiles of the intranasal and parenteral formulations were similar. Intranasal and intramuscular metoclopramide showed similar efficacy in the control of acute emesis induced by moderately emetogenic chemotherapy in 12 patients. Intranasal metoclopramide 80mg significantly reduced the frequency of acute vomiting in 43 patients receiving highly emetogenic chemotherapy. A pilot study suggested that intranasal metoclopramide, with or without dexamethasone, may reduce cisplatin-induced delayed emesis. In a randomised crossover trial in 40 patients, intranasal metoclopramide or oral metoclopramide, both with dexamethasone, were equally effective in the control of delayed emesis induced by moderately-emetogenic chemotherapy. One 30 patient study suggests that intranasal metoclopramide has similar efficacy to oral metoclopramide in the treatment of functional dyspepsia. A non-significant trend to reducing postoperative nausea and vomiting has been seen in two trials of intranasal metoclopramide. Intranasal metoclopramide caused minor irritation of the nasal membrane and unpleasant taste in some patients, but was otherwise well tolerated. None of the more serious extrapyramidal effects sometimes associated with metoclopramide were reported. PMID- 10473025 TI - Zolmitriptan: a review of its use in migraine. AB - Zolmitriptan is a selective serotonin 5-HT1B/1D receptor agonist ('triptan'). Its efficacy and tolerability have been assessed in a number of randomised, placebo controlled, double-blind trials in large numbers of adults with moderate to severe migraine attacks. Oral zolmitriptan 2.5 and 5mg has a rapid onset of action (significant headache relief is observed at 45 minutes) and efficacy is sustained in most patients who respond at 2 hours. The drug is significantly more effective than placebo as measured by a number of parameters including 2-hour headache response rates and pain-free response rates. Other symptoms of migraine, including nausea, photophobia and phonophobia are also alleviated with zolmitriptan. Zolmitriptan is effective in the treatment of migraine associated with menses and migraine with aura. There is some evidence to support the use of zolmitriptan in patients with migraine who have had a poor response to previous therapy. The efficacy of zolmitriptan appears to be maintained, with no tachyphylaxis, following repeated administration for multiple attacks of migraine over a prolonged period of time, with high headache response rates reported over all attacks. In comparison with placebo, the incidence of persistent migraine headache is reduced by zolmitriptan and recurrent migraine headache occurs less frequently with the active treatment. Zolmitriptan has also demonstrated efficacy in the treatment of persistent and/or recurrent migraine headache. For relief of migraine headache, zolmitriptan 5mg had similar efficacy to sumatriptan 100mg for a single attack, but generally was more effective than sumatriptan 25 and 50mg for multiple attacks, in single trials. The incidence of recurrent headache with zolmitriptan was similar to that with sumatriptan. Zolmitriptan is generally well tolerated with most adverse events being mild to moderate, transient and resolving without intervention or the need for treatment withdrawal. The most common adverse events with zolmitriptan therapy are asthenia, heaviness other than that of the chest or neck, dry mouth, nausea, dizziness, somnolence, paraesthesia, warm sensation, tightness, vasodilation and chest pain. CONCLUSION: Zolmitriptan is effective across a wide range of migraine subtypes, maintains efficacy when used in the long term and is generally well tolerated. Further clinical experience is necessary to define the position of zolmitriptan among other currently or soon to be available selective 5-HT1B/1D receptor agonists. However, on the basis of available data, zolmitriptan should emerge as a useful treatment option in the management of patients with moderate to severe migraine. PMID- 10473024 TI - Cidofovir: a review of its use in cytomegalovirus retinitis in patients with AIDS. AB - Cidofovir is an antiviral nucleotide analogue with significant activity against cytomegalovirus (CMV) and other herpesviruses. The drug is indicated for the treatment of CMV retinitis, a sight-threatening condition, in patients with AIDS. Cidofovir has a long intracellular half-life which allows for a prolonged interval (2 weeks) between maintenance doses. In contrast, other intravenous treatment options for patients with CMV retinitis (i.e. ganciclovir and foscarnet) must be administered on a daily basis. The efficacy of intravenous cidofovir has been demonstrated in patients with AIDS and previously untreated CMV retinitis in multicentre randomised trials, and in a dose-finding study of cidofovir in patients with AIDS and previously treated relapsing CMV retinitis. Clinical trials have been relatively small (n < or = 100 patients) and no studies have been conducted directly comparing intravenous cidofovir with the more established intravenous agents, ganciclovir or foscarnet. Indirect comparisons of clinical trial data suggest that intravenous cidofovir may have similar efficacy to intravenous ganciclovir or foscarnet in delaying progression of CMV retinitis. However, such comparisons must be made with caution because of potential differences in patient populations, data analysis techniques and interobserver variability in the masked assessment of retinal photographs. Nevertheless, intravenous cidofovir offers a less intrusive administration regimen than intravenous ganciclovir or foscarnet because of its prolonged dosage interval. Since therapy is life-long, patients receiving daily intravenous ganciclovir or foscarnet (but not cidofovir) usually require an indwelling central venous catheter and are therefore at increased risk of serious infection. The relatively long dosage interval for cidofovir may also have favourable implications in terms of overall treatment costs and patient quality of life, although specific data are very limited. Potentially irreversible nephrotoxicity is the major treatment limiting adverse event associated with intravenous cidofovir in patients with AIDS-related CMV retinitis. Anterior uveitis/iritis has been reported frequently with intravenous cidofovir in postmarketing reports and a small number of patients have developed hypotony. Other treatment options for CMV retinitis are also associated with serious adverse events, and selection of pharmacotherapy will depend on a number of factors including retinitis lesion characteristics, patient quality-of-life issues and efficacy and tolerability profiles of available therapies. CONCLUSION: Although the extent of its use may be limited by its adverse event profile, cidofovir offers a useful addition to the limited number of drugs available for the treatment of CMV retinitis in patients with AIDS. PMID- 10473027 TI - Group sequential procedures for repeated events data with frailty. AB - In this article we consider the design aspects of group sequential trials with recurrent study endpoints, where the subjects are from a heterogeneous population. The usual procedures of sequential analysis based on the "independent increments" property are no longer valid due to the heterogeneity of the study subjects, as pointed out by Cook and Lawless. We focus on the aspect of study planning and consider a special model (Poisson process with frailty) to investigate the effects of frailty (or subject heterogeneity). We prescribe the valid procedures and provide convenient tables for computing stopping boundaries and sample sizes. The methodology is illustrated through an example of an animal experiment with recurrent tumors, using procedures such as group sequential tests and repeated confidence intervals. Minimal cost analysis is considered for determining the optimal combination of study duration and sample size. PMID- 10473026 TI - Topical imiquimod: a review of its use in genital warts. AB - Imiquimod is a topically active immunomodulatory agent that is formulated as a 5% cream for application by the patient. It is the first agent of its class, the immune response modifiers, to be used in the treatment of genital warts. In immunocompetent patients with genital warts, imiquimod stimulates the production of interferon-alpha and various other cytokines, and has indirect antiviral activity. In randomised, double-blind, vehicle-controlled clinical trials, complete clearance of warts occurred in 37 to 50% of immunocompetent patients with genital warts treated with imiquimod 5% cream 3 times a week for up to 16 weeks; partial clearance of warts (defined as a reduction in wart area of > or = 50%) was observed in 76% of recipients of imiquimod 5% cream. Rates of complete or partial clearance of warts were significantly higher in patients who applied imiquimod 5% cream 3 times a week than in recipients of imiquimod 1% or vehicle cream, each applied 3 times a week. A between-gender difference in clinical response to imiquimod 5% cream has been reported, with female patients experiencing higher rates of complete clearance of warts than males. Recurrence(s) of > or = 1 wart occurred in 13 to 19% of immunocompetent patients in whom complete clearance of warts had been achieved with imiquimod 5% cream. Imiquimod 5% cream also shows some clearance of warts in immunosuppressed HIV infected patients with genital warts. Preliminary results of a vehicle-controlled study showed that the rate of partial clearance of warts (defined as a reduction in baseline wart area of >50%) [38%] was significantly higher with imiquimod 5% cream than with vehicle cream; however, the rate of complete clearance was not significantly higher than with vehicle cream. Imiquimod 5% cream is generally well tolerated by immunocompetent and HIV-infected patients. Local skin reactions (mainly mild or moderate), including erythema, itching and burning, are the most commonly reported adverse events, occurring in < or = 67% of patients applying imiquimod 5% cream 3 times a week. The incidence of adverse events is lower in patients applying the cream 3 times a week than with daily application. The incidence of systemic adverse events with imiquimod 5% cream (applied daily or 3 times a week) is similar to that of vehicle cream. The tolerability profile of imiquimod cream appears favourable compared with that of podophyllotoxin. CONCLUSION: Imiquimod 5% cream is a new therapeutic option for patients with genital warts. It produces clearance rates broadly similar to those of other treatment approaches and rates of wart recurrence compare favourably with those reported for established treatments. In contrast to most alternative treatment strategies. which are administered in the physician's office, imiquimod cream is a self-administered therapy for outpatient use. PMID- 10473028 TI - Empirical power for distribution-free tests of incomplete longitudinal data with applications to AIDS clinical trials. AB - The design of AIDS clinical trials is of growing importance. These studies tend to be longitudinal and typically involve missing data. HIV-1 RNA is a common endpoint for these studies and is inherently non-normal, although viral load can be measured only within certain bounds, resulting in censored data. We compared several analysis methods, both univariate and multivariate, on the basis of empirical power and provide an illustrative example of data from a controlled clinical trial. Simulated viral load data demonstrate that methods adjusting for baseline data have power increasing with increasing positive intrasubject correlation expected with this type of data. Several summary measures considered have power compatible with multivariate tests. PMID- 10473029 TI - Guidelines for measurement validation in clinical trial design. AB - In the process of designing a clinical trial, the accuracy and precision of an endpoint is of critical importance in being able to determine valid results. In the creation and subsequent testing of the validity of an endpoint, it is desirable to show that on repeated measurements the endpoint can be measured precisely, and that it is reproducible with not only itself, but with any "gold standard" that can assess accuracy. Short of having this gold standard, we rely on showing that the endpoint is reliable. Agreement is the extent to which the measurement of the variable of interest yields the same results (consistency) on repeated trials. Thus the more consistent the results given by repeated measurements, the more reliable the measuring procedure. In this article, we examine the sensitivity to location and scale shift of different methods for assessing agreement. These tools include Pearson's correlation coefficient, the intraclass correlation coefficient, the concordance correlation coefficient, the Bradley-Blackwood procedure, and the within patient coefficient of variation. Our simulation studies showed that there are situations wherein the various methods for assessing agreement give conflicting results. On the basis of the results of the simulation studies, there are three important components to consider when deciding whether two sets of measurements are in agreement or not. The first component is the degree of linear relationship between the two sets; the second is the amount of bias as represented by the difference in the means; and the third is the difference between the two variances. The purpose of this article is to help interpret numerical results from the measures of agreement and to establish a criterion or range of values for each agreement measure that we consider to indicate agreement. We clarify what is meant by agreement by placing the measures in context based on scale and location shifts. Furthermore, we present guidelines for assessing agreement or endpoint validation when there are plans to design a clinical trial, and we give an actual example from a recent study. PMID- 10473030 TI - Modeling the covariance structure in pharmacokinetic crossover trials. AB - Pharmacokinetic studies of drug and metabolite concentrations in the blood are usually conducted as crossover trials, especially in phases I and II. A longitudinal series of measurements is collected on each subject within each period. However, much of the dependence among such observations, within and between periods, is generally ignored in analyzing this type of data. Usually, only a random coefficient model is fitted for the parameters in the nonlinear mean function, along with allowing the variance to depend on the mean so that it changes over time. Here, we develop models to allow more fully for the structure of the crossover study. We introduce two levels of variance components, for the subjects and for the periods within subjects, and also an autocorrelation within periods. We also retain the time-varying variance, using a separate variance function for this, different from that for the mean. We apply this model to a phase I study of the drug flosequinan and its metabolite. This drug was developed for the treatment of heart failure. Because the metabolite also exhibits an active pharmacologic effect, study of both the parent drug and the metabolite is of interest. We find that the autocorrelation is the element in the covariance structure that most improves the fit of the model but that two levels of variance components can also be necessary. PMID- 10473031 TI - A method for estimating and testing area under the curve in serial sacrifice, batch, and complete data designs. AB - The extent of drug availability is often measured by the area under the concentration-time curve. In animal studies, experimental constraints can limit the number of observations available on each animal. Estimation of area under the curve and its standard error are straightforward when each animal is measured at each time point. Bailer and Nedelman et al., have described techniques for estimating the area under the curve and its standard error when each animal is measured once. Yeh has described a technique for the hybrid case where animals are measured more than once, but not at all time points. We describe a method for estimating area under the curve and its standard error which is applicable to all three types of designs. We give formulas for testing treatment differences, including dose trends and dose proportionality, in area under the curve for designs containing an arbitrary number of treatments. A jackknife estimator is also described. PMID- 10473032 TI - On testing equivalence of three populations. AB - We consider analysis of clinical trials in which the objective is to show that three populations are equivalent. Equivalence is defined in terms of delta, the maximum difference in population means; a one-sided hypothesis test of delta is considered. We provide the distribution of the maximum pairwise difference in sample means, and we use this distribution to find critical values for tests of size 0.100, 0.050, 0.025, and 0.010. When standard errors are not equal among the three treatments, a simple adjustment is proposed to control the type I error rate. These tests are applied to studying the equivalence of three binomial proportions. Test-based confidence intervals are discussed. Two examples illustrate the proposed methods. PMID- 10473033 TI - Bioequivalence review for drug interchangeability. AB - To monitor the performance of the approved generic copies of a brand-name drug, we propose some methods in assessing bioequivalence among generic copies and the brand-name drug, and among generic copies themselves, using data from several bioequivalence studies adopting the standard 2 x 2 crossover design without carryover effects. We propose a meta-analysis method that increases statistical power when the between-subject variability is not large. A nonmeta-analysis is also considered. A numerical example of applying both methods is presented for illustration. PMID- 10473034 TI - The distribution of the maximum likelihood estimator in up-and-down experiments for quantal dose-response data. AB - Standard maximum likelihood logistic or probit regression has been used in biopharmaceutical practice for inference about tolerance threshold distributions in situations where subjects (patients) have been allocated doses according to an up-and-down design. For example, a steeper dose-response curve than expected was reported in one such study. This article demonstrates that the maximum likelihood estimator systematically and considerably exaggerates the regression parameter with moderately large sample sizes. Thus a probable explanation for finding a steeper curve than expected is the method used to analyze the experiment, that is, the bias in the maximum likelihood estimator. An additional consequence of this bias is that the mean/median/ED50 are estimated with a misleading precision. In particular, confidence intervals are much too narrow. As a conclusion, we warn against conventional logistic or probit regression in combination with up-and down designs. PMID- 10473035 TI - Extending Goodman-Kruskal's gamma to the comparison of ordered treatments in multicenter clinical trials. AB - In a randomized, multicenter clinical trial setting, the treatments may consist of increasing doses of a drug and placebo and the response variable may be ordinal (e.g., physician's global evaluation of treatment effectiveness). Within each center (e.g., hospital), patients are randomly assigned to treatments (rows) such that the row totals are fixed and the rows form a product-multinomial sample of the ordinal response variable. Gamma, a measure of ordinal association in two way contingency tables, and its asymptotic standard error can be estimated from the data in each center. We use these independent estimates of gamma for testing the hypothesis of homogeneity of gammas, controlling for center effect. If this hypothesis is not rejected, the within-center estimates of gamma can be combined to form a common gamma across the centers. PMID- 10473036 TI - Letter to the editor regarding "A power study of a sequential method of p-value adjustment for correlated continuous endpoints". PMID- 10473037 TI - Vertebrate limb development and malformations. AB - To understand limb abnormalities it is necessary to understand how the limb develops. The limb is the organ whose development is probably best understood. The limbs develop from small protrusions (the limb buds) that arise from the body wall of the embryo. Positioning and patterning the limb involves cellular interactions both between the ectoderm surrounding the limb bud and between the mesenchymal cells that form the core of the limb bud. As the limb grows out the cells acquire a positional value that relates to their position in the bud with respect to all three axes, proximo-distal, antero-posterior, and dorso-ventral. These positional values largely determine how the cells will develop such as what sort of cartilaginous elements they will form. The positional value of the cells is acquired in the progress zone at the tip of the growing bud. The time spent in the progress zone may determine the positional values along the proximo-distal axis, that is the formation of, for example the humerus, then the radius and ulna. Loss of the progress zone due to damage to the overlying apical ridge leads to truncations, and this progress zone model can also account for the effects of thalidomide. Position along the antero-posterior axis such as the character of the digits is by a signal from the polarizing region at the posterior margin of the limb and involves the signaling protein Sonic hedgehog. A signal from the dorsal ectoderm specifies the dorso-ventral axis. Hox genes that are transcription factors are expressed both along the body axis and in a complex pattern in the limb and may record positional value. Human mutations in these genes lead to limb abnormalities. Muscle cells have a separate origin from the cartilaginous cells and those that form connective tissue and tendons, and they migrate into the bud from the somites and are patterned by the connective tissue. Cell death separates the digits. PMID- 10473038 TI - Definition of the human N-myc promoter region during development in a transgenic mouse model. AB - The N-myc oncogene directs organogenesis, and gene amplification is associated with aggressive forms of neuroblastoma, a common malignant tumor in children. N myc is expressed in fetal epithelium, and expression decreases markedly postnatally. To localize sequences responsible for directing expression, we have analyzed the human N-myc promoter. We noted previously that N-myc promoter regions 5' to exon 1 directed reporter gene expression in all cell lines, including those without detectable N-myc transcripts. However, when promoter constructs included 3' exon 1 and the 5' portion of intron 1, reporter activity was detected only when there was expression of the endogenous gene. To determine the role of this "tissue-specific region" in directing expression during development, we generated transgenic mice carrying N-myc promoter lacZ minigenes that contained 5' N-myc promoter elements alone or the promoter linked to the 3' exon 1/5' intron 1 tissue-specific region. Animals lacking the tissue-specific exon 1/intron 1 region showed beta-galactosidase expression in the CNS, but expression was not observed in other organs in which endogenously derived N-myc transcripts were seen. Within the CNS, transgene expression was seen mainly in the olfactory system and was not observed in other areas in which expression of the murine gene has been noted. In contrast, no transgene expression was observed in any of the animals carrying the tissue-specific exon 1/intron 1 region. Thus, sequences that direct expression within the olfactory system were contained within our 5' promoter transgene, whereas sequences that guide the ubiquitous expression of N-myc during organogenesis lie outside the regions studied here. Finally, the exon 1/intron 1 region seems to act in a dominant fashion to repress expression in the CNS from the immediate 5' N-myc promoter. PMID- 10473039 TI - Why is erythropoietin present in human milk? Studies of erythropoietin receptors on enterocytes of human and rat neonates. AB - Erythropoietin receptors (Epo-R) are expressed on cells in the small bowel of human fetuses, but their function has not been defined. We hypothesized that intestinal Epo-R are present postnatally, and that recombinant erythropoietin (rEpo) would increase enterocyte migration and decrease cytokine-induced apoptosis. We used reverse transcriptase-polymerase chain reaction and immunohistochemistry to evaluate the presence of Epo-R mRNA and protein in rat intestinal epithelial cells (IEC-6), and in postnatal human and rat bowel. The effect of rEpo on rates of cell migration and proliferation were established in IEC-6 cells by using cell counting and incorporation of bromodeoxyuridine. To determine whether rEpo affects response to injury, cells were pretreated with rEpo, then were damaged with 25 or 50 ng/mL tumor necrosis factor-alpha plus 2.5 microg/mL cycloheximide. Cell death was determined by colorimetric bioassay. We found that Epo-R mRNA and protein were expressed by IEC-6 cells and by enterocytes of postnatal rat and human small bowel. Cells that had been exposed to 0.05 or 5.00 U/mL rEpo migrated faster than did the controls (p < 0.05), but no difference was noted in cell proliferation. Treatment of IEC-6 cells with rEpo before or at the time of injury resulted in a lower percentage of cell death, and this effect was neutralized by anti-Epo antibody. We conclude that Epo-R is expressed in enterocytes postnatally in rats and humans. Recombinant Epo increases the rate of migration of IEC-6 cells and decreases cytokine-induced apoptosis. These studies suggest that Epo within human milk has actions on neonate's intestinal function. PMID- 10473040 TI - The effects of the estrogen receptor blocker, Faslodex (ICI 182,780), on estrogen accelerated bone maturation in mice. AB - Sex steroids accelerate bone maturation, but it is believed that estrogen action is needed for terminal epiphyseal fusion. In this study, we investigated the effects of a new estrogen-blocking agent, Faslodex (ICI 182,780), on estrogen accelerated skeletal maturation in immature mice. On day-of-life 2 through 8, mice pups received either estradiol (5 microg/100 g body weight), Faslodex (100 microg/100 g body weight), a combination of Faslodex + estradiol, or vehicle alone. Skeletal maturation was assessed with a scoring system based on the size and appearance of epiphyseal plates in the forepaw and the lumbar spine. Estradiol caused acceleration of bone maturation in our mouse model (p < 0.05). Faslodex blocked the effect of estrogen, such that the mice receiving Faslodex + estradiol did not vary significantly from controls. Faslodex may prove useful in the treatment of patients with diseases causing rapid skeletal maturation, such as precocious puberty. PMID- 10473041 TI - Cerebral hypothermia is not neuroprotective when started after postischemic seizures in fetal sheep. AB - Prolonged cerebral hypothermia is neuroprotective if started within a few hours of hypoxia-ischemia. However, delayed seizure activity is one of the major clinical indicators of an adverse prognosis after perinatal asphyxia. The aim of this study was to determine whether head cooling delayed until after the onset of postasphyxial seizures may still be neuroprotective. Unanesthetized near-term fetal sheep in utero received 30 min of cerebral ischemia induced by bilateral carotid artery occlusion. Eight and one-half hours later, they received either cooling (n = 5) or sham cooling (n = 13) until 72 h after the insult. Intrauterine cooling, induced by circulating cold water through a coil around the fetal head, was titrated to reduce fetal extradural temperature from 39.4+/-0.1 degrees C to between 30 and 33 degrees C. Cerebral ischemia led to the delayed development of intense epileptiform activity from 6 to 8 h postinsult, followed by a marked secondary rise in cortical impedance (a measure of cytotoxic edema) and in carotid blood flow. Cerebral cooling markedly attenuated the secondary rise in impedance and reduced carotid blood flow (p < 0.001). After 5 d recovery, there was no significant difference in loss of parietal EEG activity relative to baseline in the hypothermia compared with the control group (-12.5+/-1.4 versus 15.2+/-1.2 dB, mean +/- SEM, NS) or in parasagittal cortical neuronal loss (82+/ 9 versus 90+/-5%, NS). In conclusion, delayed prolonged head cooling begun after the onset of postischemic seizures was not neuroprotective. These data highlight the importance of intervention in the latent phase, after reperfusion but before the onset of secondary injury. PMID- 10473042 TI - Age-dependent differences in the relationship between plasma and brain extracellular fluid concentrations of magnesium after MgSO4 infusions in miniswine. AB - Magnesium is a potential neuroprotective agent in the treatment of head injury and ischemia whose efficacy is likely determined by increases in brain extracellular fluid (ECF) magnesium, which in turn depends on its concentration in plasma. The objectives of this study were to: 1) examine the effects of increasing plasma magnesium concentration ([Mg]plasma) to 4-6 mM on brain ECF magnesium concentration ([Mg]ECF) and 2) determine whether maturational changes occur in the transfer of magnesium into brain ECF for newborn and more mature (approximately 1 month old) miniswine. Increases in [Mg]plasma by systemic administration of MgSO4 resulted in similar maximal elevations in brain [Mg]ECF for both age groups (193+/-76% versus 253+/-106% of control for newborn and 1 month-old miniswine, respectively). Calculations of half-lives (t1/2) for the increase and decrease in magnesium concentration (t1/2 uptake and t1/2 clearance) were used to characterize magnesium kinetics in plasma and brain ECF. Plasma magnesium uptake was shorter in 1-month-old (t1/2 = 11.1+/-0.9 min) compared with newborns (12.9+/-1.7 min, p < 0.05). The faster increase in [Mg]plasma probably contributed to a faster uptake of brain [Mg]ECF in 1-month-old compared with newborn swine (t1/2 uptake = 27.9+/-12.8 versus 46.0+/-20.9 min, respectively, p < 0.05). Although plasma magnesium clearance was shorter in 1-month-old swine compared with newborn (t1/2 = 34.3+/-7.0 versus 74.7+/-33.7 min, respectively, p < 0.05), the clearance of magnesium from the brain ECF was similar for each age group. Reductions in blood pressure and heart rate occurred during hypermagnesemia and were similar in each age group. This study shows that acute elevations in [Mg]plasma to 4-6 mM result in similar relative increases in brain [Mg]ECF for both newborn and 1-month-old miniswine. However, there are maturational differences, as demonstrated by the faster rate of magnesium uptake into the ECF observed in the older miniswine. PMID- 10473043 TI - Cerebral intracellular lactic alkalosis persisting months after neonatal encephalopathy measured by magnetic resonance spectroscopy. AB - We have found that cerebral lactate can be detected later than 1 month of age after neonatal encephalopathy (NE) in infants with severe neurodevelopmental impairment at 1 y. Our hypothesis was that persisting lactate after NE is associated with alkalosis and a decreased cell phosphorylation potential. Forty three infants with NE underwent proton and phosphorus-31 magnetic resonance spectroscopy at 0.2-56 wk postnatal age. Seventy-seven examinations were obtained: 25 aged <2 wk, 16 aged > or = 2 to < or = 4 wk, 25 aged > 4 to < or = 30 wk, and 11 aged > 30 wk. Neurodevelopmental outcome was assessed at 1 y of age: 17 infants had a normal outcome and 26 infants had an abnormal outcome. Using univariate linear regression, we determined that increased lactate/creatine plus phosphocreatine (Cr) was associated with an alkaline intracellular pH (pHi) (p < 0.001) and increased inorganic phosphate/phosphocreatine (Pi/PCr) (p < 0.001). This relationship was significant, irrespective of outcome group or age at time of study. Between outcome groups, there were significant differences for lactate/Cr measured at < 2 wk (p = 0.005) and > 4 to < or = 30 wk (p = 0.01); Pi/PCr measured at < 2 wk (p < 0.001); pHi measured at < 2 wk (p < 0.001), > or = 2 to < or = 4 wk (p = 0.02) and > 4 to < or = 30 wk (p = 0.03); and for N acetylaspartate/Cr measured at > or = 2 to < or = 4 wk (p = 0.03) and > 4 to < or = 30 wk (p = 0.01). Possible mechanisms leading to this persisting cerebral lactic alkalosis are a prolonged change in redox state within neuronal cells, the presence of phagocytic cells, the proliferation of glial cells, or altered buffering mechanisms. These findings may have implications for therapeutic intervention. PMID- 10473044 TI - Biphasic edema after hypoxic-ischemic brain injury in neonatal rats reflects early neuronal and late glial damage. AB - Magnetic resonance imaging with diffusion- and T2-weighted imaging and 31P magnetic resonance spectroscopy was used to investigate the relationship between development of brain edema and alterations of the brain energy metabolism after hypoxia-ischemia (HI) brain injury in 7-d-old rats. The results were correlated with histologic examinations at various times during recovery up to 5 d. Moderate HI, induced by right common carotid artery ligation and subsequent exposure to 8% O2 for 90 min, produced a cytotoxic edema of 52+/-9% brain volume and depressed the ratio of phosphocreatine to inorganic phosphate from 1.43+/-0.21 to 0.11+/ 0.09. Within 1 h of reoxygenation, the edema decreased to 4+/-2% of brain volume, demarcating the core of the lesion. At 5 h of recovery, a secondary cytotoxic edema together with a newly developing vasogenic edema expanded again, reaching its maximal extent of 45+/-10% brain volume at around 24 h. The ratio of phosphocreatine to inorganic phosphate recovered slowly, reaching 1.12+/-0.27 around 13 h. Thereafter it declined again in a manner analogous to the observations made in human newborns after severe perinatal asphyxia, reaching trough values of 0.48+/-0.22 around 24 h after HI. At the cellular level, the vast majority of neuronal death occurred before 15 h. Subsequently, strong glial activation lasted 2-3 d after HI. At 5 d, a cystic infarction of 35+/-12% brain volume was found. We conclude that the biphasic evolution of brain edema and energy metabolism reflects early neuronal and late glial damage in response to moderate HI injury. Therefore, the secondary energy breakdown reflects glial activation and subsequent glial death. PMID- 10473045 TI - Survival of guinea pig pups in hyperoxia is improved by enhanced nutritional substrate availability for glutathione production. AB - The imbalance between high oxidant loads and immature antioxidant defenses is associated with long-term complications of prematurity. Glutathione is a central element among the antioxidants. Depletion of pulmonary glutathione accelerates the development of oxygen-induced lung injury in neonatal animal models. After the observation that newborn infants exposed to oxygen have low glutathione levels, a study was designed to test the hypothesis that in neonates from a species susceptible to oxygen toxicity, the lethal effect of hyperoxia is related to a low availability of substrates for glutathione production rather than an impairment in synthetic activity. One-day-old guinea pigs, randomly assigned to room air or oxygen (>95%), were fed by their mothers (n = 16) or i.v. by dextrose (n = 14) or by total parenteral nutrition (TPN, n = 20). After 3 d, glutathione and activities of enzymes involved in maintaining intracellular glutathione levels were determined in lungs and liver. The lethal effect of oxygen (p < 0.05) observed in animals without TPN was not related to glutathione depletion, as oxygen induced a 33% increase in lung glutathione, positively correlated (r2 = 0.35) with enhanced synthesis. With TPN, the animals were protected against the lethal effects of hyperoxia and lung glutathione increased by 67% in oxygen. The results suggest that the glutathione demand by the lungs in the presence of an oxidant stimulus was met by the increased (p < 0.001) hepatic production supported by TPN. Under hyperoxic conditions, early nutritional support is of vital importance. PMID- 10473046 TI - The role of nitric oxide in dilating the fetal ductus arteriosus in rats. AB - Prostaglandin E is a major dilator of the fetal ductus arteriosus (DA), but the role of nitric oxide in fetal ductal dilation has not been established. We studied the effects of a potent nitric oxide synthase inhibitor, Nomega-nitro-L arginine methyl ester (L-NAME), on the fetal DA in rats. L-NAME was injected into the dorsum of pregnant rats, and fetal DA was studied 4 h later with a rapid whole body freezing method. The inner diameters of the DA and the main pulmonary artery were measured on a freezing microtome. The inner diameter ratio of DA to main pulmonary artery (DA/PA) was 1.02+/-0.03 (mean +/- SEM; number of fetuses [n], 21) in normal near-term fetuses. The effect of prostaglandin synthesis inhibition was studied after orogastric administration of indomethacin to pregnant rats. In near-term rats on the 21st day of gestation (term, 21.5 d), a large dose of L-NAME (100 mg/kg) caused only mild ductal constriction, with DA/PA reduced to 0.83+/-0.05 (n = 20). Indomethacin (1 mg/kg) caused moderate ductal constriction, and DA/PA was decreased to 0.65+/-0.05 (n = 21). Combined administration of L-NAME (10 mg/kg) and indomethacin (1 mg/kg) caused severe ductal constriction, with DA/PA of 0.26+/-0.03 (n = 16). In preterm rats on the 19th day of gestation, a moderate dose of L-NAME (10 mg/kg) caused severe ductal constriction, with a DA/PA of 0.32+/-0.05 (n = 24). Indomethacin (1 mg/kg) alone caused only mild ductal constriction, with DA/PA 0.86+/-0.02 (n = 16). In conclusion, prostaglandin has a major role and nitric oxide has a minor role in dilating the DA in the near-term fetal rat. In contrast, nitric oxide has a major role and prostaglandin has a minor role in dilating the DA in preterm fetal rats. PMID- 10473047 TI - FK506 alters sarcoplasmic reticulum calcium release in neonatal piglet cardiac myocytes. AB - Tacrolimus (FK506) is a potent immunosuppressive drug that, when complexed to a family of immunophilin proteins known as FK binding proteins, inhibits calcineurin in T lymphocytes. Although i.v. use of FK506 in pediatric transplant recipients has been linked to development of cardiomyopathies, its mechanism of cardiotoxicity has not been examined in a neonatal animal model. In our study the effects of FK506 were investigated in cardiac myocytes isolated from newborn piglets. The peak amplitude of electrically triggered fura-2 Ca2+ transients was increased in FK506-treated myocytes, but Ca2+ transient duration and baseline fura-2 Ca2+ ratios were not altered. 45Ca2+ uptake by digitonin-lysed piglet cells decreased at pCa < or = 6.0, indicating that sarcoplasmic reticulum efflux channels were leaky. The results suggest that elevated release of sarcoplasmic reticulum Ca2+ during systole contributes to cardiotoxic effects observed in children. PMID- 10473048 TI - Precocious pubarche, dyslipidemia, and low IGF binding protein-1 in girls: relation to reduced prenatal growth. AB - An increasing series of pediatric endocrinopathies and metabolic anomalies has been recognized as related to reduced prenatal growth. We have tested whether the association of precocious pubarche (PP), dyslipidemia, and low serum IGF binding protein-1 in girls is also related to reduced prenatal growth. Fasting serum lipids, lipoproteins, and IGFBP-1 concentrations were measured in 187 girls (83 without PP and 104 with PP; mean age, 11.8 y; range, 5-18 y) with known birthweight and gestational age, the latter being transformed into birthweight SD scores. Birthweight SD scores of girls with PP were lower than those of girls without PP. Within the group of PP girls, those with dyslipidemia and low IGFBP-1 had lower (p < 0.0001) birth-weight SD scores (-2.02+/-0.23; mean +/- SEM) than those with normal lipids, lipoproteins, and IGFBP-1 (-0.37+/-0.15), whereas girls with an intermediate number of abnormalities had intermediate birthweight SD scores (-0.80+/-0.18). In conclusion, dyslipidemia and low serum IGFBP-1 in girls with PP were found to be related to reduced prenatal growth, an observation pointing to the prenatal origin of these metabolic abnormalities. PMID- 10473049 TI - Changes of serum allopregnanolone levels in the first 2 years of life and during pubertal development. AB - Allopregnanolone is the best characterized among neurosteroids, and its role in the control of neuroendocrine axes has attracted increasing interest recently. However, there is no available information about circulating levels of allopregnanolone during infancy, childhood and puberty. We studied two groups of children: 1) those aged between 0 and 2 y (n = 72), and 2) those aged between 6 and 18 y, at different Tanner's stages (n = 82). In each of these patients, serum allopregnanolone, progesterone, cortisol, and dehydroepiandrosterone levels were evaluated after informed consent; allopregnanolone was measured by RIA after acid extraction on cartridge. There was no significant variation of serum allopregnanolone levels either in male and female children during the first 2 y of life. Furthermore, although serum dehydroepiandrosterone levels showed a significant decrease, inversely correlated with age of the children (p < 0.01), serum cortisol and progesterone levels showed a significant age-related increase during the first 2 y of life. Cortisol and allopregnanolone levels were positively correlated (p < 0.01). During puberty, we observed a progressive increase in serum allopregnanolone levels in both boys and in girls, which were higher at Tanner' s stage IV-V (0.7+/-0.01 nM; mean +/- SEM) than at stages I-II (0.32+/-0.02 nM; p < 0.01); mean levels were significantly higher at puberty than in the first 2 y of life (p < 0.01). Furthermore, during puberty, serum progesterone and dehydroepiandrosterone levels also increased progressively with age in both boys and girls. Allopregnanolone and dehydroepiandrosterone levels were positively correlated throughout puberty. The present results indicate that serum allopregnanolone levels do not change during the first 2 y of life but increase during pubertal development, suggesting that this steroid may be involved in the adaptive neuroendocrine mechanisms related to puberty. PMID- 10473050 TI - The somatostatin analogue octreotide inhibits neuroblastoma growth in vivo. AB - Neuroblastoma, a neural crest-derived childhood tumor of the sympathetic nervous system, may in some cases differentiate to a benign ganglioneuroma or regress due to apoptosis. However, the majority of neuroblastomas are diagnosed as metastatic tumors with a poor prognosis despite intensive multimodal therapy. The neuropeptide somatostatin (SOM) has been shown to inhibit neuroblastoma growth and induce apoptosis in vitro. Therapeutic effects of SOM analogues are dependent on tumor expression of high-affinity receptors. In the present study, human neuroblastoma SH-SY5Y cells were grown as xenografts in nude rats. In vivo SOM receptor expression in the xenografts was identified using scintigraphy with 111In-pentetreotide. Rats were randomized to treatment with the long-acting SOM analogue octreotide (10 microg s.c. every 12 h), 13-cis-retinoic acid (4 mg orally every 24 h), or vasoactive intestinal peptide (40 microg s.c. every 24 h) and compared with controls. Tumor volume was assessed every second day and tumor weight after 10-12 d. Octreotide treatment inhibited neuroblastoma growth significantly with reduced tumor volumes at 10 and 12 d compared with untreated controls (mean 3.56 and 4.24 versus 6.48 and 8.01 mL, respectively; p < 0.01). Also, tumor weights after 10-12 d were reduced in octreotide-treated animals (n = 8, median weight 2.90 g, range 1.67-5.57 g) compared with untreated rats (n = 14, 7.54 g, 1.65-10.82 g, p = 0.005). Serum IGF-I decreased significantly over time both in rats treated with octreotide and in untreated controls. It is concluded that treatment with the SOM analogue octreotide may significantly decrease neuroblastoma tumor growth in vivo. Further studies are warranted to establish the role of SOM analogues in the treatment of children with unfavorable neuroblastoma. PMID- 10473051 TI - Distribution of granulocyte colony-stimulating factor (G-CSF) and G-CSF-receptor mRNA and protein in the human fetus. AB - Granulocyte colony-stimulating factor (G-CSF) promotes neutrophil proliferation, differentiation, and survival. It acts by binding to specific cell-surface receptors (G-CSF-R), which are expressed on cells of granulocytic lineage, human endothelial cells, and placenta. It has been postulated that the administration of recombinant G-CSF (rG-CSF) to preterm neonates might be useful in treating infections or in reducing nosocomial infections. Whereas it is known that G-CSF and G-CSF-R are present in the developing fetal bone marrow and liver, no information is available as to the existence or distribution of nonhematopoietic G-CSF-R in other tissues of the developing human fetus. We hypothesized that G CSF and its receptor might be expressed in various fetal tissues, as has been shown for other growth factors such as erythropoietin and fibroblast growth factor. Therefore, we studied the anatomical distribution of mRNA-encoding G-CSF and G-CSF-R, using RT-PCR and in situ RT-PCR in a variety of human fetal tissues ranging from 8 to 24 weeks postconception. The cellular distributions of the corresponding proteins were determined by immunohistochemistry. Both G-CSF and G CSF-R were present in nearly every organ and tissue examined, but in discrete cellular localizations. G-CSF-R in kidney and intestine underwent changes in anatomical distribution with fetal development. These results indicate that G-CSF and G-CSF-R have wide anatomical expression in the developing human fetus. PMID- 10473052 TI - Effects of oxytocin treatment early in pregnancy on fetal growth in ad libitum fed and food-restricted rats. AB - The effects of oxytocin on fetal and placental growth and on maternal weight gain and accumulation of body fat were studied in ad libitum-fed and food-restricted (receiving 70% of the food intake of the ad libitum-fed group) pregnant rats. Further, a possible role of the IGF axis in mediating oxytocin-induced changes was assessed. Pregnant rats were injected subcutaneously once a day during gestational d 1-5 with saline or oxytocin (1 mg/kg). Ad libitum-fed oxytocin treated pregnant rats had higher circulating levels of IGF-I, larger placentas, fetuses, and newborn pups and contained less body fat at the end of pregnancy. In food-restricted dams, oxytocin-treatment had no effect on fetal and placental growth. Additionally, food restriction attenuated the normal increase in IGF binding protein-3 protease proteolysis during pregnancy. The results show that oxytocin may affect maternal adaptations to pregnancy and stimulate fetal growth. We suggest that this effect may be mediated by increased IGF-I in ad libitum-fed animals, whereas food restriction may block this effect by resulting in low levels of circulating IGF-I and by attenuating the pregnancy-associated increase in IGF binding protein-3 protease activity and, thereby, further compromise IGF bioavailability. PMID- 10473053 TI - Synthesis of mevalonate pathway lipids in fibroblasts from Zellweger and X-linked ALD patients. AB - Fibroblasts were cultured to determine the involvement of peroxisomes in cholesterol and dolichol synthesis. For this purpose, the behavior of cells from patients with Zellweger syndrome, with X-linked adrenoleukodystrophy, and from nondiseased control subjects was studied. Cells both after pretreatment with mevinolin and without pretreatment were incubated in a medium containing [3H] mevalonate. In fibroblasts from patients with peroxisomal defects, the cholesterol content and mevalonate incorporation into cholesterol were decreased by 10-20% in comparison with control cells. Mevinolin pretreatment decreased the incorporation rate of [3H]-mevalonate into cholesterol but increased the labeling of ubiquinone and dolichol both in diseased and control cells. Squalene synthase activity was unchanged, whereas the activity of farnesyl-pyrophosphate synthase was increased in the diseased states. The results show that in patients with peroxisomal deficiency neither the amount nor the rate of synthesis of cholesterol and dolichol is reduced to any greater extent. PMID- 10473054 TI - Corticotropin-releasing hormone as adrenal androgen secretagogue. AB - The regulation of adrenarche is one of the enigmas of pediatric endocrinology. Adrenarche is thought to be governed by a dual control mechanism in which an adrenal androgen secretagogue acts upon a zona reticularis primed by ACTH. We hypothesized that corticotropin-releasing hormone (CRH) may serve as adrenal androgen secretagogue. We tested the concept by infusing either saline or human (h) CRH (1 microg/kg/h in saline) over 3 h, after overnight dexamethasone pretreatment, into eight young men within a randomized, cross-over study design. Serum ACTH and dehydroepiandrosterone-sulfate were measured once hourly; DHEA, androstenedione and 17-hydroxy-progesterone were determined at baseline and after 3 h of saline/hCRH infusion. ACTH levels remained unaltered during saline infusion and average ACTH responses amounted to 13 pg/mL (3.3 pmol/L) during hCRH infusion. Neither saline nor hCRH infusion altered 17-hydroxy-progesterone levels. Serum dehydroepiandrosterone-sulfate rose swiftly within 3 h of hCRH infusion and remained unchanged after saline (mean increase 37 versus 1%; p < 0.01). On average, serum DHEA doubled and androstenedione tripled during hCRH infusion, although no changes were observed during saline infusion (p < 0.01). In conclusion, CRH appears to have the capacity to act as adrenal androgen secretagogue. We suggest that the enigma of adrenarche may have an elegant solution, with CRH and ACTH coupled in sequence at the hypothalamic-pituitary level, and in parallel within the zona reticularis, just as they presumably are within the fetal adrenal, which is exposed to CRH of placental origin. PMID- 10473055 TI - Ties that bind. PMID- 10473056 TI - Retrograde drilling of osteochondral lesions of the medial talar dome. AB - Sixteen patients (16 ankles) with symptomatic osteochondral lesions of the medial talar dome were treated arthroscopically with percutaneous retrograde drilling through the sinus tarsi. The surgical technique allows preservation of intact articular cartilage, in contrast to traditional methods. All patients treated with this technique during a 24-month period were included in the study. Patient age ranged from 16 to 44 years (mean, 33 years). Follow-up ranged from 19 to 38 months (mean, 24 months). A staging system based on magnetic resonance imaging examination was used to grade the lesions preoperatively. Outcome was evaluated using the American Orthopaedic Foot and Ankle Society Ankle-Hindfoot Scale. Preoperative scores ranged from 35 to 75 points, with a mean of 53.9 points. Postoperative American Orthopaedic Foot and Ankle Society scores ranged from 48 to 100 points, with a mean of 82.6 points. Mean improvement was 25 points. There were no surgical complications. Short-term results were comparable to results reported with other available techniques. PMID- 10473057 TI - Analysis of the heel pad fat in rheumatoid arthritis. AB - The heel fat pad is organized, both in structure and in composition, to bear the stresses and strains of normal activities and to permit pain-free weightbearing. The fatty acid composition of heel pads in 11 patients with rheumatoid arthritis, a disease process frequently associated with heel fat pad atrophy, was analyzed using gas-liquid chromatography and was compared with that of patients without systemic disease. The heels of patients with rheumatoid arthritis demonstrated a significant change in the composition of saturated fatty acids when compared with heels of nonrheumatoid patients. This composition reflects an increased fat viscosity, which decreases the ability of the heel to absorb and dissipate the energy generated during ambulation. This factor could cause degeneration of the heel septal system, with resulting fat pad atrophy. PMID- 10473058 TI - Ankle arthrodesis in rheumatoid arthritis using an intramedullary nail with fins. AB - We describe an intramedullary nail with fin-like longitudinal ridges that we have developed for arthrodesis of the ankle in rheumatoid arthritis. Four fins with sharp tips were attached to the distal part of a cylindrical nail to stabilize the tibiotalar and subtalar joints. We used this nail in 15 feet of 15 patients with rheumatoid arthritis who were followed for an average of 34.9 months. Postsurgery, 13 patients were allowed to bear weight immediately, as tolerated, without immobilization. By 3 weeks, these patients were able to bear weight fully. Solid fusion of the ankle joint in an acceptable position and good clinical results were obtained in all patients. The only complications were two cases of delayed wound-healing. PMID- 10473059 TI - Ankle fusion in a high risk population: an assessment of nonunion risk factors. AB - Between July 1992 and April 1996, 88 ankle fusions were performed at our institution. Sixty-seven of these had adequate follow-up for evaluation for union of the fusion, including adequate records and/or radiographs. The average age of patients was 43 years. There were 37 men and 24 women. The charts were reviewed to determine what level of trauma had resulted in posttraumatic arthritis (low energy, high energy, or open fracture). Alcohol use, drug abuse, diabetes, peripheral vascular disease, psychiatric history, smoking, or technical problems were also assessed. A chi-square analysis was used to evaluate the statistical significance. Nineteen of sixty-seven ankle fusions progressed to nonunion (28%). Eighty-five percent of the patients had posttraumatic arthritis. Among 17 patients with a history of open trauma, nine patients developed a nonunion (P < 0.03). A trend toward significance was noted for patients who were smokers, drank alcohol, had diabetes, had a psychiatric disorder, or used illegal drugs. Even with current techniques, this study demonstrates that a high risk population in a trauma center is at risk for nonunion after an ankle fusion caused by multiple risk factors, including a history of open trauma, tobacco use, alcohol use, illegal drug use, a history of psychiatric disorders, or diabetes. PMID- 10473060 TI - Stress fractures and tibial cortical hypertrophy after tibiotalocalcaneal arthrodesis with an intramedullary nail. AB - Twelve patients underwent successful tibiotalocalcaneal arthrodesis using a locked intramedullary nail with posterior moldable bone grafting. Two of the patients developed a nondisplaced stress fracture around the proximal interlocking screws, which responded to immobilization at 8 and 15 months after surgery. In 7 of 12 patients, > or =2 mm of cortical hypertrophy was noted between the proximal interlocking screws along the medial and/or lateral aspect of the tibia. Although intramedullary nailing can achieve satisfactory tibiotalocalcaneal arthrodesis, the stress risers in the tibial diaphyses do result in a stress reaction in a significant number of patients. PMID- 10473061 TI - Ankle arthroplasty in patients younger and older than 50 years: a prospective series with long-term follow-up. AB - One hundred consecutive cases treated with ankle arthroplasty for osteoarthritis (OA) or rheumatoid arthritis were followed prospectively and annually for up to 15 years. Survivorship analysis was performed, with the endpoint being prosthesis revision or change to arthrodesis. Patients who were younger than 50 years at the first implantation constituted one group (group A, 30 ankles). The other group (group B, 70 ankles) consisted of patients aged 50 years or older at the first implantation. All patients were assessed clinically according to the Kofoed Ankle Score. The distribution of OA/rheumatoid arthritis in group A was 18/12, and in group B it was 43/27 (not significant). The median age in group A was 46 years (range, 22-49 years), and in group B it was 63 years (range, 51-83 years). In group A, one case was revised, and three cases were converted to arthrodesis after a median of 5 years (range, 5-9 years). In group B, four cases were revised, and four cases were converted to arthrodesis after a median of 5.5 years (range, 2-8 years). The results of cases with traumatic OA did not differ between groups A and B. It was concluded that the results of ankle arthroplasty were of equal quality in patients younger than 50 years and those who were older. PMID- 10473062 TI - Relative movement of the navicular bone during normal walking. AB - The purpose of this study was to determine the pattern and magnitude of navicular bone (NB) movement during walking as well as the relationship between dynamic NB and rearfoot movement. The angle of rearfoot and displacement of the NB was recorded in 106 subjects using the 6D Research electromagnetic tracking system. The relative change in the height of the NB between foot flat and heel-off was 7.9 mm. The NB seems to undergo significant vertical as well as medial displacement during the stance phase of normal walking. This motion is also correlated with rearfoot motion during walking. PMID- 10473063 TI - Relationship between foot pronation and rotation of the tibia and femur during walking. AB - The purpose of this study was to test the hypothesis that the magnitude and timing of peak foot pronation would be predictive of the magnitude and timing of peak rotation of tibia and femur. Thirty subjects who demonstrated a wide range of pronation participated. Three-dimensional kinematics of the foot, tibia, and femur segments were recorded during self-selected free walking trials using a six camera VICON motion analysis system. Regression analysis demonstrated that the magnitude and timing of peak pronation was not predictive of the magnitude and timing of tibial and femoral rotation. The lack of a relationship between peak foot pronation and the rotation of the tibia and femur is contrary to the clinical hypothesis that increased pronation results in greater lower extremity rotation. It would seem, therefore, that the relationship between foot pronation and rotation of the lower extremity segments should be assessed on a patient-by patient basis. PMID- 10473064 TI - Elevated plantar pressure and ulceration in diabetic patients after panmetatarsal head resection: two case reports. AB - Panmetatarsal head resection (variously called forefoot arthroplasty, forefoot resection arthroplasty, the Hoffman procedure, and the Fowler procedure) was developed for the relief of pain and deformity in rheumatoid arthritis. Although there are successful retrospective series reported in the literature, such an approach is not supported by carefully designed controlled trials. This procedure has also been advocated by some for the relief of plantar pressure in diabetic patients who are at risk for plantar ulceration. The efficacy of the procedure in this context is not supported by existing pressure measurements on rheumatoid arthritis patients in the literature, which has tended to show that although pain relief is obtained, the procedure results in elevation of forefoot pressure. Case reports are described of two patients (three feet) with sensory neuropathy who presented to our clinic 1 to 2 years after panmetatarsal head resections had been performed. Peak plantar pressures in these feet during first step gait were above the 99th percentile and outside the measuring range of the device used (EMED SF platform; NOVEL Electronics Inc., St. Paul, MN). Both patients had also experienced plantar ulcers subsequent to the surgery. Combining the information on patients with rheumatoid arthritis (RA) with that from our two case studies, we conclude that panmetatarsal head resection does not necessarily eliminate focal regions of elevated plantar pressure. PMID- 10473065 TI - Fibular nonunion and equinovarus deformity secondary to posterior tibial tendon incarceration in the syndesmosis: a case report after a bimalleolar fracture dislocation. AB - A 39-year-old woman sustained a grade II open bimalleolar fracture-dislocation of the left ankle. Six months after an ORIF of these fractures was performed, she presented with a nonunion of the distal fibula fracture and with a fixed hindfoot equinovarus and forefoot adduction deformity. At surgery for repair of the fibular nonunion, the posterior tibial tendon (PTT) was found to be entrapped in the posterior tibiotalar joint, with a portion of the tendon interposed between the tibia and the fibula in the area of the posterior syndesmosis. After extrication of the PTT, the hindfoot varus and forefoot adduction deformity were corrected. To our knowledge, this is the first case report in the English literature of a missed PTT syndesmotic entrapment that resulted in a fibular nonunion and in a fixed foot deformity after an open bimalleolar ankle fracture dislocation. PMID- 10473066 TI - Divot sign: a new observation in anterior impingement of the ankle. AB - Anterior impingement of the ankle results from osteophyte formation on the anterior edge of the distal tibia. Classically, subsequent degeneration results in an osteophyte forming on the anterior neck of the talus. This results in limited dorsiflexion and impingement seen in the ankles of athletes. Recent clinical observation in high performance soccer players has revealed a previously unrecognized pattern of a localized divot forming in the talar neck in place of the osteophyte. This accepts the tibial osteophyte during dorsiflexion, which is therefore not limited in these patients. PMID- 10473067 TI - Synovial osteochondromatosis of the retrocalcaneal bursa: a case study. AB - A 42-year-old man presented with synovial osteochondromatosis (SO) of the retrocalcaneal bursa with intraosseous lesions in the calcaneum, leading to significant pain and consequent disability. Simple x-rays, computed tomography, and magnetic resonance imaging were performed preoperatively. The patient has remained asymptomatic, clinically and radiographically, without recurrent lesions for 18 months postoperatively. PMID- 10473068 TI - Use of the lamina spreader for reduction of the subtalar joint during hindfoot surgery. PMID- 10473069 TI - Rank order analysis of tibial plafond fractures: does injury or reduction predict outcome? PMID- 10473070 TI - Dihydropyrimidine dehydrogenase activity: prognostic partner of 5-fluorouracil? PMID- 10473071 TI - Molecular detection of micrometastases and circulating tumor cells in solid tumors. AB - The detection of circulating tumor cells and micrometastases may have important prognostic and therapeutic implications. Because their numbers can be very small, these tumor cells are not easily detected using conventional methods. In the last decade, molecular techniques have been widely used for the detection of occult tumor cells. The objective of this report is the application of these molecular tools to solid tumors. A systematic review of all related English-language articles published in the last 32 years was performed. The molecular detection of occult tumor cells can be accomplished by PCR amplification of tumor-specific abnormalities present in the DNA or mRNA of malignant cells. The other main PCR strategy for the detection of CTC and micrometastases involves amplification of tissue-specific mRNA. This latter method was often applied to solid tumors, whereas the former was occasionally used. PCR was shown to be superior to conventional techniques in detecting occult tumor cells, allowing the identification of 1 malignant cell mixed with 1 to 10 million normal cells. In some reports, PCR is shown to be a strong predictor of outcome. The molecular detection of circulating tumor cells and micrometastases in solid tumors can be accomplished using highly sensitive PCR assays. The central question of whether PCR reliably predicts relapse and survival remains unanswered for many types of solid tumor. If PCR-based assays are found to be a reliable tool, they will likely have a major impact on the management of these malignancies. PMID- 10473072 TI - Detection of tumor messenger RNA in the serum of patients with malignant melanoma. AB - Serum RNases are known to be elevated in patients with cancer. Consequently, it is not clear whether human mRNA with sufficient integrity as to permit reverse transcription-PCR (RT-PCR) amplification is detectable in serum. We examined serum from six patients with malignant melanoma for human tyrosinase mRNA using RT-PCR. Serum from 20 normal volunteers served as controls. Tyrosinase mRNA could be demonstrated in serum from four of the six melanoma patients with detection by gel electrophoresis and confirmation by blotting amplified product to a tyrosinase-specific probe. The serum remained tyrosinase mRNA positive, even if passed through a 0.45 microm filter prior to RNA extraction, indicating that the mRNA was extracellular at the time of extraction. Tyrosinase mRNA could not be detected in any control serum (0 of 20 individuals). The presence and integrity of amplifiable RNA was confirmed in all serum specimens (patients and controls) by RT-PCR amplification of c-abl mRNA. Amplifiable RNA could be demonstrated regardless of whether serum was freshly drawn or stored frozen for several years. We conclude that human mRNA can be extracted and amplified from serum. The ability to amplify tumor mRNA from serum may have important utility in cancer diagnostics and monitoring. PMID- 10473073 TI - Tissue microarrays for gene amplification surveys in many different tumor types. AB - Gene amplifications are common in many different tumor types and may confer diagnostic, prognostic, or therapeutic information for patient management. Tedious experiments are often required to determine which tumor types have amplifications of a specific oncogene. To facilitate rapid screening for molecular alterations in many different malignancies, a tissue microarray consisting of samples from 17 different tumor types was generated. Altogether, 397 individual tumors were arrayed in a single paraffin block. To determine whether results from the literature can be reproduced on minute tissue samples (diameter, 0.6 mm), amplification of three extensively studied oncogenes (CCND1, CMYC, and ERBB2) was analyzed in three fluorescence in situ hybridization experiments from consecutive sections cut from the tissue microarray. Amplification of CCND1 was found in breast, lung, head and neck, and bladder cancer, as well as in melanoma. ERBB2 was amplified in bladder, breast, colon, stomach, testis, and lung cancer. CMYC was amplified in breast, colon, kidney, lung, ovary, bladder, head and neck, and endometrial cancer. These results confirm and even extend existing data in the literature on such amplifications. In summary, we applied three fluorescence in situ hybridization experiments to analyze amplifications of three oncogenes in three x 397 tumors within a week. This demonstrates the power of using minute arrayed tissue specimens for tumor screening. PMID- 10473074 TI - Suicide prodrugs activated by thymidylate synthase: rationale for treatment and noninvasive imaging of tumors with deoxyuridine analogues. AB - Most tumors are resistant to therapy by thymidylate synthase (TS) inhibitors due to their high levels of TS. Instead of inhibiting TS, we hypothesized that it was possible to use this enzyme to activate suicide prodrugs (deoxyuridine analogues) to more toxic species (thymidine analogues). Tumors with high levels of TS could be particularly sensitive to deoxyuridine analogues because they would be more efficient in producing the toxic methylated species. Furthermore, the accumulation of methylated species within tumors could be visualized externally if a tracer dose of the deoxyuridine analogue was tagged with an isotope, preferably a positron emitter, such as 18F. Higher accumulation of isotope indicates higher activity of TS and lower sensitivity of the tumor to TS inhibitors, but perhaps more sensitivity to therapy with deoxyuridine analogues as suicide prodrugs. 2'-F-ara-deoxyuridine (FAU) was used as a prototype to demonstrate these concepts experimentally. FAU readily entered cells and was phosphorylated, methylated, and subsequently incorporated into cellular DNA. Among different cell lines, FAU produced varying degrees of growth inhibition. Greater DNA incorporation (e.g., for CEM and U-937 cells) was reflected as increased toxicity. FAU produced less DNA incorporation in Raji or L1210 cells, and growth rate was minimally decreased. As the first demonstration that cells with high levels of TS activity can be more vulnerable to therapy than cells with low TS activity, this preliminary work suggests a new therapeutic approach for common human tumors that were previously resistant. Furthermore, it appears that the TS activity of tumors could be noninvasively imaged in situ by tracer doses of [18F]FAU and that this phenotypic information could guide patient therapy. PMID- 10473075 TI - Inhibition of matrix metalloproteinase-2 expression and bladder carcinoma metastasis by halofuginone. AB - Matrix metalloproteinase-2 (MMP-2) plays a critical role in tumor cell invasion and metastasis. Inhibitors of this enzyme effectively suppress tumor metastasis in experimental animals and are currently being tested in clinical trials. MMP-2 transcriptional regulation is a part of a delicate balance between the expression of various extracellular matrix (ECM) constituents and ECM degrading enzymes. Halofuginone, a low-molecular-weight quinazolinone alkaloid, is a potent inhibitor of collagen type alpha1 (I) gene expression and ECM deposition. We now report that expression of the MMP-2 gene by murine (MBT2-t50) and human (5637) bladder carcinoma cells is highly susceptible to inhibition by halofuginone. Fifty percent inhibition was obtained in the presence of as little as 50 ng/ml halofuginone. This inhibition is due to an effect of halofuginone on the activity of the MMP-2 promoter, as indicated by a pronounced suppression of chloramphenicol acetyltransferase activity driven by the MMP-2 promoter in transfected MBT2 cells. There was no effect on chloramphenicol acetyltransferase activity driven by SV40 promoter in these cells. Halofuginone-treated cells failed to invade through reconstituted basement-membrane (Matrigel) coated filters, in accordance with the inhibition of MMP-2 gene expression. A marked reduction (80-90%) in the lung colonization of MBT2 bladder carcinoma cells was obtained after the i.v. inoculation of halofuginone-treated cells as compared with the high metastatic activity exhibited by control untreated cells. Under the same conditions, there was almost no effect of halofuginone on the rate of MBT2 cell proliferation. These results indicate that the potent antimetastatic activity of halofuginone is due primarily to a transcriptional suppression of the MMP-2 gene, which results in a decreased enzymatic activity, matrix degradation, and tumor cell extravasation. This is the first description, to our knowledge, of a drug that inhibits experimental metastasis through the inhibition of MMP-2 at the transcriptional level. Combined with its known inhibitory effect on collagen synthesis and ECM deposition, halofuginone is expected to exert a profound anticancerous effect by inhibiting both the primary tumor stromal support and metastatic spread. PMID- 10473076 TI - A Phase I and pharmacokinetic study of TNP-470 administered weekly to patients with advanced cancer. AB - A Phase I study of angiogenesis inhibitor TNP-470 was conducted in patients with advanced cancer. TNP-470 (25-235 mg/m2) was administered i.v. over 4 h once a week to patients who had solid tumors refractory to the best available treatment or with a high risk of recurrence and who had normal renal, hepatic, and hematological function and no evidence of coagulopathy. The aims of the study were to determine the maximum tolerated dose, dose-limiting toxicities (DLTs), and the pharmacokinetics of TNP-470 given on a once-weekly schedule. Thirty-six patients, ages 23-75 (median, 54 years), with an Eastern Cooperative Oncology Group performance status of 0-2 were treated. The number of patients at each dose level (mg/m2) were 6 (25), 3 (50), 3 (75), 3 (100), 3 (133), 12 (177), and 6 (235). The principal toxicities of TNP-470 were dizziness, lightheadedness, vertigo, ataxia, decrease in concentration and short-term memory, confusion, anxiety, and depression, which occurred at doses of 133, 177, and 235 mg/m2. Two patients treated at 235 mg/m2 experienced DLT in the form of grade III cerebellar neurotoxicity after 6 weeks of treatment. Overall, these neurological symptoms were dose-related, had an insidious onset, progressively worsened with treatment, and resolved completely within 2 weeks of stopping the drug. One patient with malignant melanoma had stabilization of the previously growing disease for 27 weeks while on the treatment. Two patients, one with adenocarcinoma of the colon and the other with a soft tissue sarcoma, had no clinically detectable disease but were at high risk for recurrence at the initiation of treatment and received 13 months and > 3 years of treatment, respectively, with no evidence of disease recurrence. The remaining patients had progression of their disease after 1-6 months of treatment. The mean plasma half-life (t(1/2)) of TNP-470 and its principal metabolite, AGM-1883, were extremely short (harmonic mean, t(1/2) of 2 and 6 min, respectively) with practically no drug detectable in the plasma by 60 min after the end of the infusion. MII, an inactive metabolite, had a considerably longer t(1/2) of approximately 2.6 h. Mean peak TNP-470 concentrations were > or = 400 ng/ml at doses > or = 177 mg/m2. On the basis of this study, the maximum tolerated dose of TNP-470 administered on a weekly schedule was 177 mg/m2 given i.v over 4 h. The principal DLT was neurotoxicity, which appeared to be dose-related and was completely reversible. On the basis of the short plasma t(1/2) of TNP-470, exploration of a prolonged i.v. infusion schedule is warranted. PMID- 10473077 TI - Thymidylate synthase protein expression in advanced colon cancer: correlation with the site of metastasis and the clinical response to leucovorin-modulated bolus 5-fluorouracil. AB - Recently, we have demonstrated that thymidylate synthase (TS) protein expression predicts for the clinical response to a regimen of infusional 5-fluorouracil (5FU) in advanced colorectal cancer patients. Previous studies by other groups that showed a correlation between TS gene expression and response to the fluoropyrimidine also involved infusional regimens. Considering the putatively different mechanism of action of bolus compared with continuous infusion of 5FU, the aim of the present study was to test whether the correlation between TS expression and the clinical response to 5FU is valid for bolus regimens. A secondary aim was to compare TS levels between liver metastases and abdominal recurrences from colon cancer, because these sites have a distinctly different responsiveness to 5FU chemotherapy. The study population consisted of 41 patients (25 males and 16 females; median age, 60 years) with unresectable metastatic or recurrent colon cancer, homogeneously treated with 5FU (420 mg/m2 i.v., days 1-5) and leucovorin (20 mg/m2 i.v., days 1-5); cycles were repeated every 28 days. Twenty-seven patients (66%) showed high levels of TS expression as defined by TS scores equal to 3 and 4. The proportion of cases with high levels of TS expression was significantly higher in abdominal recurrences (18 of 22, 82%) compared with liver metastases (9 of 19, 47%; P = 0.02). Intratumoral TS protein expression was inversely correlated with response to chemotherapy (response rate: 7 of 14, 50%, versus 0 of 27 in patients with low and high levels of TS expression, respectively; P = 0.0001). These results confirm that the level of TS protein expression predicts for response to 5FU, even with a bolus schedule. The higher TS levels observed in abdominal compared with liver metastases may account for their different responsiveness to 5FU chemotherapy. Immunohistochemical quantitation of TS protein levels may thus allow us to change the therapeutic approach to advanced colorectal cancer from a general to an individual treatment strategy at a time when new non TS-targeted drugs have become available for this disease. PMID- 10473078 TI - Pharmacokinetic study of S-1, a novel oral fluorouracil antitumor drug. AB - S-1 is a novel oral fluorouracil antitumor drug that combines three pharmacological agents: tegafur (FT), which is a prodrug of 5-fluorouracil (5 FU); 5-chloro-2,4-dihydroxypyridine (CDHP), which inhibits dihydropyrimidine dehydrogenase (DPD) activity; and potassium oxonate (Oxo), which reduces gastrointestinal toxicity. Phase I and early Phase II clinical trials have already been completed. On the basis of the results of these trials, 80 mg/m2/day, given daily in two divided doses after breakfast and supper, a 28-day consecutive oral regimen is recommended. In this study, we investigated the pharmacokinetics of 5-FU, intact FT, CDHP, and Oxo, after administration of S-1, at a standard dose of 80 mg/m2/day, in advanced cancer patients. Twelve patients were recruited to the study; 5 patients with gastric cancer, 4 with colorectal cancer, and 3 with breast cancer. Among them, analysis was conducted on 12 patients for single administration and on 10 patients for consecutive administration. The initial dose of S-1 for each patient was determined according to his/her body surface area (BSA) as follows: for BSA < 1.25 m2, 80 mg/body/day; for 1.25 m2 < or = BSA < 1.5 m2, 100 mg/day; and for 1.5 m2 < or = BSA, 120 mg/day. For single administration, half of the standard dose was used. For 28-day consecutive administration, the standard dose was given daily in two divided doses. The average single dose per BSA was 35.9 mg/m2 (31.7-39.7 mg/m2). Pharmacokinetic parameters of plasma 5-FU were as follows: Cmax, 128.5 +/- 41.5 ng/ml; Tmax, 3.5 +/- 1.7 h; AUC(0-14), 723.9 +/- 272.7 ng x h/ml; and T(1/2), 1.9 +/- 0.4 h. In the 28-day consecutive regimen, there were no fluctuations in pharmacokinetics nor any drug accumulation. Because the pharmacokinetics of orally administered S-1 is almost similar to that of continuous i.v. infusion of 5-FU, we concluded that S-1 may improve patients' quality of life. PMID- 10473079 TI - Life-threatening toxicity in a dihydropyrimidine dehydrogenase-deficient patient after treatment with topical 5-fluorouracil. AB - In humans, 80-90% of an administered dose of 5-fluorouracil (5-FU) is degraded by dihydropyrimidine dehydrogenase (DPD; EC 1.3.1.2), the initial rate-limiting enzyme in pyrimidine catabolism. Cancer patients with decreased DPD activity are at increased risk for severe toxicity including diarrhea, stomatitis, mucositis, myelosuppression, neurotoxicity, and, in some cases, death. We now report the first known cancer patient who developed life-threatening complications after treatment with topical 5-FU and was shown subsequently to have profound DPD deficiency. RT-PCR and genomic PCR methodologies were used to identify a G to A mutation in the GT 5' splicing recognition sequence of intron 14, resulting in a 165-bp deletion (corresponding to exon 14) in this patient's DPD mRNA. Immunoprecipitation and Western blot analysis were then used to demonstrate that the aberrant DPD mRNA is translated into a nonfunctional DPD protein that is ubiquitinated. We conclude that the presence of this metabolic defect combined with topical 5-FU (a drug demonstrating a narrow therapeutic index) results in the unusual presentation of life-threatening toxicity after treatment with a topical drug. These data further suggest that degradation by the ubiquitin proteosome-mediated system plays a role in the elimination of the DPD protein. PMID- 10473080 TI - Drug-administration sequence does not change pharmacodynamics and kinetics of irinotecan and cisplatin. AB - In this study, 11 patients with solid tumors were randomized to receive irinotecan (CPT-11; 200 mg/m2) as a 90-min i.v. infusion, immediately followed by cisplatin (CDDP; 80 mg/m2) as a 3-h i.v. infusion in the first course and the reversed sequence in the second course or vice versa. No significant differences in any toxicity were observed between the treatment schedules (decrease in absolute neutrophil count, 74.7 +/- 18.3 versus 80.3 +/- 18.0%; P = 0.41). CPT-11 lactone clearance was similar to single agent data and not significantly different between study courses (60.4 +/- 17.1 versus 65.5 +/- 16.3 liter/h/m2; P = 0.66). The kinetic profiles of the major CPT-11 metabolites SN-38, SN-38 glucuronide, 7-ethyl-10-[4-N-(5-aminopentanoic acid)-1 piperidinolcarbonyloxycamptothecine (APC), and 7-ethyl-10-[4-N-(1-piperidino)-1 amino]carbonyloxycamptothecine (NPC) were also sequence independent (P > or = 0.20). In addition, CPT-11 had no influence on the clearance of nonprotein-bound CDDP (40.8 +/- 16.7 versus 50.3 +/- 18.6 liter/h/m2; P = 0.08) and the platinum DNA-adduct formation in peripheral leukocytes in either sequence (1.94 +/- 2.20 versus 2.42 +/- 1.62 pg Pt/microg DNA; P = 0.41). These data indicate that the toxicity of the combination CPT-11 and CDDP is schedule independent and that there is no mutual pharmacokinetic interaction. PMID- 10473081 TI - Overexpression of cyclooxygenase-2 in carcinoma of the pancreas. AB - The level of cyclooxygenase (COX)-2 has been investigated recently in various human carcinomas. In the present study, we examined the distribution and extent of COX-2 protein in human pancreatic tumors using immunohistochemistry. A strong expression of COX-2 protein was present in 23 of 52 (44%) pancreatic carcinomas, a moderate expression was present in 24 of 52 (46%) pancreatic carcinomas, and a weak expression was present in 5 of 52 (10%) pancreatic carcinomas. In contrast, benign tumors showed weak expression or no expression of COX-2, and only islet cells displayed COX-2 expression in normal pancreatic tissues. Overexpression of COX-2 in carcinoma tissues was also confirmed by Western blot analysis. Furthermore, consistent with the results at protein levels, reverse transcription PCR analyses indicated that COX-2 mRNA was overexpressed in 7 of 13 (54%) carcinomas, but in none of 3 benign tumors. Our findings suggest that COX-2 inhibitors might be potentially effective against pancreatic carcinomas and that COX-2 may be involved in certain biological processes in pancreatic islets. PMID- 10473082 TI - Loss of heterozygosity in epithelial cells obtained by bronchial brushing: clinical utility in lung cancer. AB - To determine whether loss of heterozygosity (LOH) could be a useful diagnostic test for lung cancer, we evaluated LOH in cells obtained from bronchial brushings. Cells from radiographically normal and abnormal lungs were obtained from 55 patients undergoing diagnostic bronchoscopy. Among 38 patients with lung cancer, LOH was present in at least one chromosomal locus in 79%, whereas cytology was positive for malignant cells in 37%. LOH was not restricted to the airway containing the tumor; fifty-three percent of the cancer patients had LOH in the contralateral lung, as did 59% of patients without lung cancer. There was an association between the extent of LOH and proximity to the cancer. The LOH score, which combined measures of fractional allelic loss and percentage of cells with allelic loss, was greater in subjects with positive cytology and on the side of the tumor. A LOH score >10 was positive in 58% of tumor-bearing lungs, in 13% of the contralateral lungs in cancer patients, and in no patients without cancer. Our results suggest that extensive and widespread allelic loss, as indicated by a high LOH score, may be diagnostic of lung cancer. Additional studies will be needed to clarify the clinical potential of using bronchial epithelial cell LOH as a biomarker and diagnostic test for lung cancer. PMID- 10473083 TI - Prognostic value of ornithine decarboxylase and polyamines in human breast cancer: correlation with clinicopathologic parameters. AB - The polyamines putrescine, spermidine, and spermine and ornithine decarboxylase (ODC), the rate-limiting enzyme in their biosynthetic pathway, play an important role in cell proliferation, differentiation, and transformation. In the present study, we have analyzed polyamine concentrations and ODC activity in samples from benign breast diseases (n = 36), benign breast tissue adjacent to the primary carcinoma (n = 19), and breast carcinoma (n = 104). ODC activity in primary carcinoma was significantly higher (2.42 +/- 0.22 nmol CO2/h g; P < 0.001) than that found in benign breast (0.62 +/- 0.15 nmol CO2/h g) or in breast tissue adjacent to the primary carcinoma (0.52 +/- 0.16 nmol CO2/h g). The total polyamine content of breast cancer tissues was higher than in benign breast diseases (704.3 +/- 38.3 nmol/g wet weight versus 295.8 +/- 27.4 nmol/g wet weight) and correlated well with ODC activity (Pearson, r = 0.42; P < 0.001). ODC activity correlated with histological grade, peritumoral lymphatic or blood vessel invasion, S-phase fraction, and cathepsin D. Total polyamine concentration increased with S-phase fraction, cathepsin D, and aneuploidy. No significant correlation was found between ODC or polyamines and tumor size, lymph node involvement, or steroid receptor status. A major finding in our study was that ODC activity was an independent prognostic factor for recurrence and death. The results indicate that the estimation of ODC activity and polyamines in human breast carcinoma might be useful to determine tumor aggressiveness and suggest that ODC may have a potential value as both a prognostic factor and a chemoprevention target in human breast cancer. PMID- 10473084 TI - Association between molecular detection of GAGE and survival in patients with malignant melanoma: a retrospective cohort study. AB - We used GAGE as a molecular marker to identify melanoma cells with metastatic potential in the peripheral blood and the bone marrow. One hundred thirty-three patients with malignant melanoma (21 clinical stage II, 74 stage III, and 38 stage IV) had a single marrow and/or blood sample drawn immediately prior to surgical resection. Simultaneous bone marrow and blood samples (85 patients), marrow-only samples (35 patients), and blood-only samples (13 patients) were examined for the presence of GAGE expression using reverse transcription-PCR. GAGE expression was associated with adverse overall patient survival, measured from the time of sampling (P = 0.01). When data were stratified for clinical stage, median survival was statistically longer among GAGE-negative patients in the stage III cohort only (P = 0.01). In a multivariate model, only GAGE positivity in blood and/or marrow and clinical stage were significant prognostic variables. It was the detection of GAGE in blood but not marrow that was associated with poor survival. The detection of blood GAGE by reverse transcription-PCR has significant adverse implications for overall survival of patients with malignant melanoma in this cohort, and it warrants further investigation. PMID- 10473085 TI - Expression of DNA topoisomerase IIalpha and topoisomerase IIbeta genes predicts survival and response to chemotherapy in patients with small cell lung cancer. AB - Drug resistance is a major problem in patients with small cell lung cancer; in fact, most die of resistant disease, despite an initial response. Several markers of drug resistance have been described in preclinical models, but the mechanism of drug resistance in lung cancer patients remains unknown. The objective of this study was to evaluate the role of the expression of a number of markers of drug resistance, proliferation, and apoptosis in relation to response to chemotherapy and survival in patients with small cell lung cancer. Tumor samples were derived from 93 previously untreated patients who were randomized in a Phase III study to receive cyclophosphamide, epirubicine, and etoposide or cyclophosphamide, epirubicine and vincristine alternating with carboplatin and etoposide. Paraffin embedded samples, derived from the primary tumor site prior to chemotherapy, were analyzed by immunohistochemistry for expression of markers implicated in drug resistance [topoisomerase (topo) IIalpha, topo IIbeta, and multidrug resistance associated protein], apoptosis (p53, p21, and bcl-2), or proliferation (Ki67). Response prediction was analyzed by chi2 test and logistic regression analysis; overall and disease-free survival curves were compared by log-rank test and Cox regression analysis. Shorter survival was observed in patients with extensive disease (P = 0.037) and poorer performance status (P = 0.028) and in patients whose tumors expressed high topo IIalpha levels (P = 0.01) and high Ki67 (P = 0.024). By multivariate analysis, the following factors were found to be predictive for worse survival: high expression levels of topo IIalpha, Ki67, and bcl-2; male sex; and extensive disease. High topo IIbeta expression was found to be predictive for lower overall and complete response rate. No relationship between apoptotic pathway markers or MRP and response to chemotherapy was observed. In conclusion, high expression of topo IIalpha was predictive of worse survival, and high expression of topo IIbeta was predictive of lower response rates. Furthermore, lower survival probability was observed in patients with bcl 2-positive tumors. Immunohistochemical assessment of these markers in diagnostic biopsies may give important prognostic information and may help selecting patients in the worse prognostic categories for new therapeutic strategies. PMID- 10473086 TI - O6-alkylguanine-DNA alkyltransferase in cutaneous T-cell lymphoma: implications for treatment with alkylating agents. AB - Mycosis fungoides is a low-grade cutaneous T-cell lymphoma. Early treatment often involves the use of topical chemotherapy such as mechlorethamine or carmustine although single-agent oral chemotherapy with alkylators is common for advanced disease. Recently, in a Phase I study of the new alkylating agent temozolomide, two mycosis fungoides patients experienced a complete response. The mechanism of resistance to alkylating drugs such as temozolomide is thought to be due to the presence in tumor cells of the DNA repair protein, O6-alkylguanine-DNA alkyltransferase (AGT). The protein mediates a reaction with the O6-position of guanine in DNA, removing the lesion and leaving guanine intact. We, therefore, examined the levels of AGT in CD4+ T lymphocytes obtained by negative antibody selection from the blood of noncancerous individuals and mycosis fungoides patients, and in paraffin-embedded sections from mycosis fungoides patch, plaque, or tumor lesions and cells from involved lymph nodes. AGT protein levels were measured by quantitative immunofluorescence microscopy using a monoclonal antibody against human AGT. Using this approach, the mean level of our positive control (AGT-expressing cells) was 84,807 molecules/nucleus; values below 5,000 molecules/nucleus are considered very low. The mean AGT level in CD4+ T lymphocytes from noncancerous and cancerous individuals was 18,618 (n = 12) and 8,593 (n = 5), respectively. The mean fraction of outliers in circulating CD4+ T lymphocytes from mycosis fungoides patients was statistically significantly lower than T cells in lymph nodes. AGT molecules/nucleus from lymph node biopsies from 8 of 10 patients showed low (< 10,000 molecules/nucleus) or undetectable levels (n = 5) of AGT. The mean AGT level from samples of mycosis fungoides patch/plaque and tumor was also low at 221 (n = 4) and 2,363 (n = 6), respectively. Surprisingly, Hut78, a mycosis fungoides T-cell lymphoma cell line, was positive for AGT activity (median: 77,700 molecules/nucleus), and Hut102--another mycosis fungoides cell line--was low (median: 5,990 molecules/nucleus). Because AGT is a primary means of cell resistance to alkylating agents, the low level of AGT in neoplastic T lymphocytes from patients with mycosis fungoides suggests that treatment with alkylating agents producing O6-alkylguanine adducts, such as carmustine or temozolomide, may produce improved clinical outcomes. PMID- 10473087 TI - Prognostic significance of perirectal lymph node micrometastases in Dukes' B rectal carcinoma: an immunohistochemical study by CAM5.2. AB - Lymph node metastasis is an important prognostic factor for rectal carcinoma, but only a few attempts at defining the relationship between lymph node micrometastases and prognosis have been made. The purpose of this study was to examine the correlation between the presence of micrometastases and prognosis in patients with rectal carcinoma. Six hundred forty-four lymph nodes were dissected from 42 patients with Dukes' B rectal carcinoma and stained immunohistochemically using a monoclonal antibody, CAM5.2, that binds cytokeratin. Clinicopathological factors, rate of recurrence, and prognosis were compared among patients with and without micrometastases. Micrometastases were detected in 19 lymph nodes (19 of 644 = 2.9%) from 9 patients (9 of 42 = 21.4%). The presence of micrometastases was not related to clinicopathological factors. There were significant differences in recurrence rates (5 of 9 versus 5 of 33, P = 0.02), relapse-free survival rates (P = 0.04), and 10-year survival rates (P = 0.03) between patients with and without micrometastases. Immunohistochemistry successfully identified micrometastatic foci in lymph nodes missed with conventional staining methods. The existence of micrometastases influenced the prognosis in patients with Dukes' B rectal carcinoma. PMID- 10473088 TI - Overexpression of cyclin D1 messenger RNA predicts for poor prognosis in estrogen receptor-positive breast cancer. AB - Cyclin D1 is a key cell cycle regulatory protein with demonstrated oncogenic activity in a variety of malignancies. Cyclin D1 mRNA and protein are overexpressed in approximately 50% of primary breast carcinomas; however, the pathophysiological consequences of increased expression remain unclear. To investigate the functional sequelae of cyclin D1 mRNA overexpression, we analyzed clinical outcome in relation to the cyclin D1 mRNA level in 253 primary breast cancer patients (median follow-up, 75 months) with particular reference to estrogen receptor (ER) status and endocrine response. Overall, with the exception of the relationship between cyclin D1 mRNA expression and the ER, cyclin D1 mRNA was not associated with other clinicopathological features such as age, menopausal status, axillary lymph node status, vascular invasion, tumor size, type, and grade. However, in patients with ER-positive tumors (n = 182), high levels of cyclin D1 mRNA were associated with increased risk of relapse (P = 0.0016), local recurrence (P = 0.025), metastasis (P = 0.019), and death (P = 0.025). In contrast, there were no clinical correlations with cyclin D1 expression in ER-negative disease (n = 71). In 33 patients who received endocrine therapy for their primary or recurrent breast cancers, there was an apparent association between a high cyclin D1 mRNA level and a shorter response duration within the ER-positive subgroup (P = 0.04). Our findings indicate that overexpression of cyclin D1 mRNA correlates with a worse prognosis within the ER positive breast cancer phenotype and may be a contributing factor to the development of endocrine resistance in ER-positive disease. PMID- 10473089 TI - Telomerase activity as a prognostic indicator in stage I non-small cell lung cancer. AB - Patients with stage I non-small cell lung cancer (NSCLC) are typically treated with surgical resection alone. However, about one-third of such patients develop disease recurrence and die within 5 years after complete resection. The ability to predict recurrence could represent an important contribution to treatment planning. This study evaluates the presence of telomerase activity in tumor cells as a predictor of disease recurrence and cancer-related death after operation for stage I NSCLC patients. The activity of the telomerase enzyme was investigated by telomeric repeat amplification protocol (TRAP) in tumors and matching normal lung tissue samples obtained from 107 consecutive operable patients with pathological stage I NSCLC. Telomerase activity was detected in 66 (62%) of the 107 tumors examined and in none of the corresponding adjacent noncancerous lung tissue samples. Correlation with pathological parameters showed that telomerase activity was associated with histopathological grade (P = 0.0135) but not with tumor size or histological type. Univariate survival curves, estimated using the method of Kaplan and Meier, defined a significant association between telomerase activity and both disease-free survival (P = 0.0115) and overall survival (P = 0.0129). In multivariate analyses, performed by Cox's proportional hazards regression models, the presence of telomerase activity was the only strong predictor of disease-free survival (P = 0.0173) and overall survival (P = 0.0187). Our data indicate that telomerase activity can be an important prognostic factor that should be considered in future prospective trials of adjuvant therapy for high-risk stage I NSCLC patients. PMID- 10473090 TI - Inactivation of the p53 pathway in prostate cancer: impact on tumor progression. AB - To determine the potential role of p53 inactivation in prostate cancer, we studied a well characterized cohort of 86 patients treated with radical prostatectomy. We analyzed patterns of p53, mdm2, and p21/WAF1 expression by immunohistochemistry. Results were then correlated with clinicopathological parameters of poor outcome, including time to PSA relapse. In addition, data were also correlated with proliferative index, as assessed by Ki67 antigen detection. p53-positive phenotype, defined as identification of nuclear immunoreactivity in > 20% of tumor cells, was observed in 6 of 86 cases (7%). An association was observed between p53-positive phenotype and decreased time to PSA relapse (P < 0.01). mdm2-positive phenotype, defined as > or = 20% of tumor cells displaying nuclear immunoreactivity, was observed in 28 of 86 cases (32.5%). mdm-2-positive phenotype was found to be associated with advanced stage (P = 0.009). p21 positive phenotype, defined as > 5% of tumor cells with nuclear immunoreactivity, was observed in 28 of 86 cases (32.5%). An association was observed between p21 positive phenotype and high Ki67 proliferative index (P = 0.002). Patients with p21-positive phenotype had a significant association with decreased time to PSA relapse (P = 0.0165). In addition, a significant association was found between p21-positive phenotype and coexpression of mdm2 (P < 0.01). Forty-three of 86 cases (50%) were found to have one or more alterations, and patients with any alteration were found to have a higher rate of PSA relapse (P < 0.01). It is our hypothesis that a pathway of prostate cancer progression involves p53 inactivation caused by mdm2 overexpression and that p21 transactivation in this setting is due to an alternative signaling system, rather than through a p53 dependent mechanism. PMID- 10473091 TI - Wild-type p53 epitope naturally processed and presented by an HLA-B haplotype on human breast carcinoma cells. AB - To broaden the clinical applicability of peptide-based immunotherapy in breast cancer, there is a need to identify further tumor-associated peptide epitopes that are specific for HLA alleles, in addition to HLA-A2. The HLA-B44 haplotype is one of the most common HLA-B haplotypes, occurring in 10-20% of the population. We performed the structural characterization of HLA class I-bound self-peptides presented by a human breast cancer cell line with a HLA-A68, A32, B40, B44 haplotype, to identify potential tumor-specific antigens. Of 13 sequenced peptides, 1 peptide had the HLA-A68 peptide binding motif and 12 peptides had the HLA-B40, B44 peptide binding motif. One of the latter peptides, FEVRVCACPG, shared 100% homology to residues 270-279 of wild-type P53 protein. Our study, thus, provides direct evidence for the natural processing and presentation of p53 epitope 270-279 by HLA-B40, B44-bearing human breast tumor cells. Epitopes spanning this region of P53 may have potential use for immunotherapy in patients expressing HLA-A2 and -B44 supertypes. PMID- 10473092 TI - Expression of plasminogen activator inhibitors 1 and 2 in lung cancer and their role in tumor progression. AB - The plasminogen activator cascade initiated by urokinase type plasminogen activator (u-PA) is involved in extracellular matrix degradation during the tumor invasion process. The plasminogen activator inhibitors 1 (PAI-1) and 2 (PAI-2) are two specific inhibitors of u-PA. We hypothesized that the balance between u PA and its two inhibitors could be disrupted to favor plasminogen activation during lung cancer progression. Using immunohistochemistry, we analyzed the pattern of expression of u-PA, PAI-1, and PAI-2 in non-small cell lung carcinomas (NSCLC) and neuroendocrine (NE) lung tumors. u-PA and PAI-1 were both detected in stromal fibroblasts and in tumor cells. In 84 NSCLCs, their epithelial expression was strongly correlated and linked to the presence of node metastasis (P = 0.008), whereas their coexpression in fibroblasts was associated with larger tumor size (P = 0.04) and advanced stages (P = 0.009). In 72 NE tumors, u-PA and PAI-1 were more frequently expressed in fibroblasts in high-grade NE tumors (SCLC and large cell NE tumors) than in low- and intermediate-grade tumors (typical and atypical carcinoids). Comparison of in situ hybridization and immunohistochemistry in 14 cases showed that PAI-1 was consistently expressed by stromal fibroblasts, although the protein was also localized in tumor cells. In contrast, the expression of PAI-2 was restricted to fibroblasts and correlated with the absence of nodal involvement (P = 0.005). Considering NE tumors, the frequency of PAI-2 expression decreased along the NE spectrum from typical carcinoids to SCLCs. These data suggest that PAI-lacts in synergy with u-PA to favor tumor invasion process and connotes aggressivity, in contrast with PAI-2, which may block u-PA-mediated proteolysis and is inversely correlated with tumor progression. PMID- 10473093 TI - The human orphan receptor PXR messenger RNA is expressed in both normal and neoplastic breast tissue. AB - The expression of PXR mRNA and a variant PXR mRNA, deleted in 111 nucleotides in the ligand-binding domain, was detected by reverse transcription-PCR amplification in both normal and neoplastic human breast tissues. The level of PXR mRNA did not differ between breast tumors and their adjacent matched normal breast tissues. However, the expression of PXR mRNA did vary among breast tumors. A statistically significant inverse relationship was found between the level of PXR mRNA expression and estrogen receptor (ER) status, as defined by ligand binding analysis. The level of PXR mRNA expression in ER+ tumors (median = 22.4, n = 15) was significantly lower (P = 0.04) than the level of PXR mRNA expression in ER-tumors (median = 46.7, n = 15). No relationship with progesterone receptor status was found. These data raise the possibility that PXR has a role in human breast tissues. PMID- 10473094 TI - Sequences homologous to the mouse mammary tumor virus env gene in human breast carcinoma correlate with overexpression of laminin receptor. AB - We previously reported that a 660-bp sequence that is homologous to the env gene of the mouse mammary tumor virus (MMTV) but not to endogenous retroviruses or to other known genes was present in 38% of human breast cancers and in some breast cancer cell lines studied (Y. Wang et al., Cancer Res., 55: 5173-5179, 1995). Here, we have investigated whether the MMTV-like sequences were associated with the clinical, pathological, and molecular parameters that have been reported to define two subsets of human breast cancers. Archival breast carcinoma samples were analyzed for four clinical parameters, obtained from patients' records, and for six pathological characteristics. Expression of c-erbB-2, p53, bcl-2, progesterone receptor, laminin receptor, and cathepsin D was detected by immunochemistry using monoclonal antibodies. PCRs were used to amplify 250 bp of the MMTV env gene-like sequence. The chi2, log-rank, and generalized Wilcoxon tests were used to analyze the data. The MMTV env gene-like sequence was detected in 37.7% of the samples. The presence of this sequence was not significantly associated with any of the pathological clinical or biological parameters studied. It did correlate, however, with expression of the laminin receptor, a marker for invasiveness and poor prognosis. This is the first phenotypic characterization of human breast cancers containing retroviral sequences. PMID- 10473095 TI - Expression of dominant-negative Ikaros isoforms in T-cell acute lymphoblastic leukemia. AB - Ikaros, a zinc finger-containing DNA-binding protein, is required for normal lymphocyte development. Germ-line mutant mice that express only non-DNA binding dominant-negative "leukemogenic" Ikaros isoforms lacking critical NH2-terminal zinc fingers develop an aggressive form of T-cell leukemia. We studied Ikaros gene expression in leukemic cells from 18 children with T-cell acute lymphoblastic leukemia (T-ALL). In each of the 18 T-ALL cases as well as JK-E6-1 and MOLT-3 cell lines, we found high-level expression of dominant-negative isoforms of Ikaros with abnormal subcellular compartmentalization patterns. Nuclear extracts from these cells failed to bind to the IKAROS-specific binding sequence in DNA. PCR cloning and sequencing confirmed that JK-E6-1 and MOLT-3 cell lines as well as leukemic cells from 9 of 10 patients with T-ALL expressed dominant-negative Ikaros isoforms Ik-4, Ik-7, and Ik-8 that lack critical NH2 terminal zinc fingers. In 6 of 10 patients, we detected a specific mutation leading to an in-frame deletion of 10 amino acids (delta KSSMPQKFLG) upstream to the transcription activation domain and adjacent to the COOH-terminal zinc fingers of Ik-2, Ik-4, Ik-7, and Ik-8. Thus, children with T-ALL express high levels of dysfunctional dominant-negative Ikaros isoforms. PMID- 10473096 TI - Minichromosome maintenance proteins as biological markers of dysplasia and malignancy. AB - Dysplasia, an intermediate stage in the progression from normal tissue to neoplasia, is defined morphologically by a loss of normal orientation between epithelial cells, with changes in cellular and nuclear shape and size. However, little is known about the functional properties of dysplastic cells, including their replicative state, largely due to a lack of available biological markers. We have used novel antibodies against minichromosome maintenance (MCM) proteins to examine the proliferative status of a range of histological lesions and to characterize dysplastic cells in functional terms. Immunoperoxidase staining was used to localize the MCM proteins, components of the prereplicative complex that is essential for initiating eukaryotic DNA replication. These proteins are down regulated in cells undergoing differentiation or quiescence and, thus, serve as specific markers for proliferating cells. In normal and some reactive tissues, MCM expression was present only in restricted proliferative compartments, consistent with our published findings in the uterine cervix. In dysplastic and malignant tissues, in contrast, MCM proteins were expressed in the majority of cells, extending to surface layers of dysplastic stratified epithelia. In carcinomas, the frequency of expression of MCM proteins showed an inverse correlation with the degree of tumor differentiation. Thus, we suggest that dysplastic cells may be characterized in functional terms as remaining in cell cycle, due to deregulation of normal controls over cell proliferation. Antibodies against MCM proteins have potential clinical applications, for example, in the assessment of tumor prognosis in histological sections and the identification of proliferating cells in clinical samples using biochemical or cytological assays. PMID- 10473097 TI - Analysis of cyclin D1 (CCND1) allelic imbalance and overexpression in sporadic human pituitary tumors. AB - Cyclin D1 plays an important role in the regulation of cell progression through G1 of the cell cycle and has been demonstrated to have oncogenic properties. Using RFLP-PCR, an A/G polymorphism within the cyclin D1 (CCND1) gene was analyzed in 151 sporadic human pituitary tumors, of which 60 were informative at this locus. Further analysis showed that in 15 of 60 (25%) tumors, there was evidence of allelic imbalance, which is indicative of gene amplification. Allelic imbalance was observed more frequently in invasive tumors (11 of 29 tumors; 38%) than in their noninvasive counterparts (4 of 31 tumors; 13%; P = 0.02). Forty-six of the tumors informative for the polymorphism were available for immunohistochemical analysis. Cyclin D1 expression (nuclear and/or cytoplasmic) was detected in 25 of 46 (54%) tumors. Of these cases, expression of nuclear cyclin D1 was detected in 9 of 46 (20%) tumors, whereas 16 of 46 (35%) tumors showed cyclin D1 staining exclusively confined to the cytoplasm. Neither nuclear staining nor cytoplasmic staining was observed in any of the normal pituitaries or in the negative control. Expression of cyclin D1 was observed in significantly more nonfunctional tumors (18 of 27 tumors; 67%) than in somatotrophinomas (7 of 19 tumors; 37%; P = 0.046). Nuclear cyclin D1 expression was observed more frequently in nonfunctional tumors (8 of 27 tumors; 30%) than in somatotrophinomas (1 of 19 tumors; 5%; P = 0.04). There was no correlation between cyclin D1 expression and tumor grade or between allelic imbalance of CCND1 and cyclin D1 expression. We conclude that amplification of CCND1 occurs in pituitary tumors and that the overexpression of cyclin D1 may be an early event in tumorigenesis. Cyclin D1 overexpression occurring in the absence of CCND1 allelic imbalance suggests that additional mechanisms responsible for deregulated cyclin D1 expression are involved in human pituitary tumorigenesis. PMID- 10473098 TI - Adenovirus-mediated p53 gene transduction inhibits telomerase activity independent of its effects on cell cycle arrest and apoptosis in human pancreatic cancer cells. AB - Evidence for a relationship between overexpression of wild-type p53 and telomerase activity remains controversial. We investigated whether p53 gene transduction could cause telomerase inhibition in pancreatic cancer cell lines, focusing on the relation of transduction to growth arrest, cell cycle arrest, and apoptotic cell death. The cells were infected with recombinant adenovirus expressing wild-type p53 or p21WAF1 at a multiplicity of infection of 100 or were continuously exposed to 10 microM VP-16, which is well known to induce apoptosis. Adenovirus-mediated p53 gene transduction caused G1 cell cycle arrest, apoptosis, and resultant growth inhibition in MIA PaCa-2 cells; the cell number 2 days after infection was 50% of preinfection value, and 13% of the cells were dead. Moreover, the transduction resulted in complete depression of telomerase activity through down-regulation of hTERT mRNA expression. In contrast, p21WAF1 gene transduction only arrested cell growth and cell cycle at G1 phase, and VP-16 treatment inhibited cell growth with G2-M arrest and apoptosis; after treatment, the cell number was 73% of pretreatment, and 12% of the cells were dead. Neither p21WAF1 gene transduction nor VP-16 treatment caused telomerase inhibition. Similar results were obtained in two other pancreatic cancer cell lines, SUIT-2 and AsPC-1. Thus, our results demonstrate that the p53 gene transduction directly inhibits telomerase activity, independent of its effects on cell growth arrest, cell cycle arrest, and apoptosis. PMID- 10473099 TI - Oral administration of the immunomodulator JBT-3002 induces endogenous interleukin 15 in intestinal macrophages for protection against irinotecan mediated destruction of intestinal epithelium. AB - We recently reported that p.o. administration of the new synthetic bacterial lipopeptide JBT-3002 can protect mice from irinotecan (CPT-11)-induced intestinal injury, but the mechanism was not known. Because interleukin-15 (IL-15) is associated with maintenance of intestinal epithelial cell integrity, we examined whether p.o. administration of JBT-3002 elevates expression of this monocyte derived cytokine. Four daily i.p. injections of 100 mg/kg CPT-11 were effective against liver metastases produced by CT-26 murine colon cancer cells, but severe damage to the intestinal epithelium and early death of the mice also resulted. Three consecutive daily p.o. doses of JBT-3002 prior to i.p. injection of irinotecan prevented the undesirable side effects of irinotecan without reducing its ability to eradicate liver metastases. Immunohistochemical analyses of the intestines of mice treated with JBT-3002 and CPT-11 demonstrated an increase in the number of dividing cells in the crypts and enhanced expression of IL-15 in lamina propria cells; the increase correlated with increased expression of the IL 15 gene as determined by semiquantitative reverse transcriptase-PCR. In vitro studies demonstrated that JBT-3002 induced expression of IL-15 in peritoneal macrophages but not in normal intestinal epithelial cells (IEC-6). Moreover, the presence of IL-15 decreased irinotecan-mediated cytotoxicity of IEC-6 epithelial cells. These data show that the p.o. administration of JBT-3002 induces expression of IL-15 by macrophages in the lamina propria, which can prevent irinotecan-induced injury to the intestinal mucosa. PMID- 10473100 TI - Efficacy of recombinant methioninase in combination with cisplatin on human colon tumors in nude mice. AB - The present treatment of colon cancer is based on 5-fluorouracil (5-FU). Despite promising results of combining leucovorin or levamisole with 5-FU, the 5-year survival rate of patients with advanced colon cancer has not increased significantly. Colon tumors in vitro have been shown previously to have an elevated requirement for methionine, suggesting a new therapeutic target. In this study, targeting the methionine dependence of colon tumors is effected by recombinant methioninase (rMETase), alone and in combination with cisplatin (CDDP). In vitro results demonstrated that CDDP and rMETase act synergistically on the human colon cancer cell line SW 620, with a combination index (CI) of 0.45, as well as on the human colon cancer cell line Colo 205 with a CI of 0.7. Human colon cancer lines HCT 15, HT 29, Colo 205, and SW 620 growing in nude mice were treated with rMETase to determine an effective dose for depletion of tumor methionine. rMETase at 15 units/g/day for 5 days depleted tumor methionine in all four tumor types to approximately 30% of untreated control. rMETase alone arrested growth of HCT 15 and HT29 in nude mice for 1 week after treatment termination. Colo 205 and SW 620 were partially arrested by rMETase. However, CDDP in combination with rMETase resulted in tumor regression of Colo 205 and growth arrest of SW 620 in nude mice. The ratio of the treated:control group (T:C) tumor weights for Colo 205 was 8% when CDDP was given on day-5, followed by treatment on days 5-9 with rMETase. This treatment schedule resulted in two of the six animals having no detectable tumor when the experiment was terminated on day 16. SW620 was resistant to CDDP alone and only partially sensitive to rMETase alone. However, when SW 620 was treated with rMETase from days-5 to -9 and CDDP on day-5, tumor growth was arrested. The results demonstrate that rMETase used simultaneously in combination with CDDP had significant antitumor efficacy in colon cancer in vitro and in vivo. The data suggest a novel and promising therapeutic approach by targeting the elevated methionine dependence of colon cancer. PMID- 10473101 TI - Decreased Src tyrosine kinase activity inhibits malignant human ovarian cancer tumor growth in a nude mouse model. AB - The Src protein tyrosine kinase is overexpressed and activated in a number of human cancers, including some human ovarian cancers. To determine whether Src activity plays a role in ovarian tumor growth, stable derivatives of the SKOv-3 human ovarian cancer cell line that exhibited reduced Src tyrosine kinase activity were generated by transfection with an antisense c-src construct. Comparison of these cell lines with parental SKOv-3 cells and stable sense c-src vector-transfected control lines revealed no phenotypic alterations in anchorage dependent proliferation, adherence, density saturation, or wound migration. However, reduction in Src activity was associated with altered cellular morphology, dramatically reduced anchorage-independent growth, and, when assessed for tumor development in a xenograft nude mouse model, diminished tumor growth. Furthermore, reduction of Src activity in the antisense c-src cell lines was associated with reduced vascular endothelial growth factor mRNA expression in vitro, and tumors derived from these cell lines displayed a phenotype indicative of abortive microvessel vascularization. These results strongly suggest that Src is involved in critical oncogenic pathways that modulate tumor growth from this ovarian cell line. Furthermore, this evidence suggests that as in other tumor systems, Src activity is required for vascular endothelial growth factor induction and angiogenic development. PMID- 10473102 TI - Androgen and epidermal growth factor down-regulate cyclin-dependent kinase inhibitor p27Kip1 and costimulate proliferation of MDA PCa 2a and MDA PCa 2b prostate cancer cells. AB - Low levels of p27Kip1 in primary prostate cancer specimens have been shown to be associated with higher rates of disease recurrence and poor rates of disease-free survival in patients with localized disease. In this study, we provide the first direct evidence showing that dihydrotestosterone (DHT), a major proliferation regulator of prostate cancer, can down-regulate p27Kip1 and stimulate cyclin dependent kinase-2 (CDK2) activity in established prostate cancer cell lines. We investigated the cooperative effects of DHT and epidermal growth factor (EGF) on the proliferation of androgen-responsive MDA PCa 2a and MDA PCa 2b prostate cancer cells. DHT and EGF each stimulated proliferation of these cells, but exposure of the cells to DHT and EGF together stimulated greater proliferation. Stimulation of cell proliferation by DHT and/or EGF was associated with increased CDK2 activity and a decreased level of p27Kip1. There seems to be a positive feedback stimulation loop between androgen-induced gene transcription and EGF stimulated signal transduction, as one could stimulate the synthesis of the receptors for the other. Dual blockade of androgen receptor function with the antiandrogen hydroxyflutamide and EGF receptor superfamily-mediated signal transduction with the anti-EGF receptor monoclonal antibody C225 and the anti HER2 receptor monoclonal antibody Herceptin significantly enhanced growth inhibition of the MDA PCa 2a cells. Our results demonstrate the importance of counteracting both androgen receptors and EGF receptors in the development of novel therapies for prostate cancer. PMID- 10473103 TI - Chlorambucil induction of HsRad51 in B-cell chronic lymphocytic leukemia. AB - Our previous studies with B-cell chronic lymphocytic leukemia (B-CLL) have suggested that one of the mechanisms of nitrogen mustard (NM) drug resistance is increased repair of drug-induced damage. We have postulated that recombination may play a crucial role in this process. The human homologue of Rad51, (HsRad51), has homology to the RecA protein in Escherichia coli, which is implicated in recombination repair and induction of DNA repair enzymes. In this report, we have examined the expression and distribution of HsRad51 protein in lymphocytes from patients with B-CLL to see whether the expression of HsRad51 is associated with NM damage to the malignant B lymphocytes, specifically chlorambucil (CLB), which is the standard alkylating agent used to treat patients with B-CLL. We have analyzed the intracellular distribution of HsRad51 protein in these lymphocytes before and after treatment with CLB by immunofluorescence. In vitro CLB treatment induces Rad51 expression, as measured by increased immunopositive staining in all CLL samples. In the CLB-resistant CLL lymphocytes, there was a linear correlation between induction of Rad51 protein at 5.4 microM CLB and the in vitro LD50 dose of CLB. Surprisingly, although it has been reported that Rad51 is induced in S phase and only 10% of cells from cell lines expressed positive immunostaining for Rad51, our CLL lymphocytes, which were not subjected to in vitro drug exposure, were 90% positive for Rad51, despite their nonproliferative state, which suggests that there is chronic activation of the protein. Our results suggest that CLB activates HsRad51-directed recombination repair and that this process may be important in NM drug-induced cytotoxicity. PMID- 10473104 TI - UFT and its metabolites inhibit the angiogenesis induced by murine renal cell carcinoma, as determined by a dorsal air sac assay in mice. AB - UFT, an anticancer agent that is composed of tegafur (FT) and uracil at a molar ratio of 1:4, is widely used in clinical practice in Japan to treat cancer patients requiring a long-term chemotherapy, and it is associated with few side effects, if any. In this study, we have evaluated the inhibitory effect of UFT against RENCA cell-induced angiogenesis by a dorsal air sac assay. Marked angiogenesis is induced by implantation of a chamber containing RENCA cells into mice. In this model, UFT showed a strong angiogenesis-inhibitory effect, whereas 5-fluorouracil (5-FU) and doxifluridine were less effective. Additional experiments revealed FT to be effective component of UFT; uracil remained ineffective in the inhibition of angiogenesis. Moreover, we have found that gamma hydroxybutyric acid and gamma-butyrolactone, the metabolites of FT, possess a potent angiogenesis inhibitory effect that is amplified when the compounds are administered by a continuous infusion. This may reflect a transition in blood concentration of each metabolite resulting from the administration of UFT. Similar results were also obtained with respect to 5-FU. It was suggested that UFT has a stronger angiogenesis-inhibitory effect than did other fluorinated pyrimidines, partly due to its pharmacokinetic properties characterized by maintaining of higher and long-lasting blood levels of 5-FU and partly due the inhibitory effects derived from gamma-hydroxybutyric acid and gamma butyrolactone, UFT-specific metabolites. PMID- 10473105 TI - Metabolic activation of dacarbazine by human cytochromes P450: the role of CYP1A1, CYP1A2, and CYP2E1. AB - Dacarbazine (DTIC), a widely used anticancer agent, is inactive until metabolized in the liver by cytochromes P450 to form the reactive N-demethylated species 5-[3 hydroxymethyl-3-methyl-triazen-1-yl]-imidazole-4-carboxamide (HMMTIC) and 5-[3 methyl-triazen-1-yl]-imidazole-4-carboxamide (MTIC). The modest activity of DTIC in the treatment of cancer patients has been attributed in part to lower activity of cytochromes P450 (P450) in humans when compared with rodents. Importantly, the particular P450 isoforms involved in the activation pathway have not been reported. We now report that the DTIC N-demethylation involved in MTIC formation by human liver microsomes is catalyzed by CYP1A1, CYP1A2, and CYP2E1. The most potent inhibitors of DTIC N-demethylation were alpha-naphthoflavone (CYP1A1 and CYP1A2), quercetin (CYP1A2), chlorzoxazone (CYP1A2 and CYP2E1), and di-sulfiram (CYP2E1). Antihuman CYP1A2 antiserum also inhibited DTIC N-demethylation. DTIC N demethylation in a panel of 10 human liver microsome preparations was correlated with the catalytic activities for CYP1A2 (ethoxyresorufin O-deethylation and caffeine N3-demethylation) in the absence of alpha-naphthoflavone and with the catalytic activities for CYP2E1 (chlorzoxazone 6-hydroxylations) in the presence of alpha-naphthoflavone. DTIC metabolism was catalyzed by recombinant human CYP1A1, CYP1A2, and CYP2E1. The Km (Vmax) values for metabolism of DTIC by recombinant human CYP1A1 and CYP1A2 were 595 microM (0.684 nmol/min/mg protein) and 659 microM (1.74 nmol/min/mg protein), respectively. The CYP2E1 Km value exceeded 2.8 mM. Thus, we conclude that (a) CYP1A2 is the predominant P450 that catalyzes DTIC hepatic metabolism; (b) CYP2E1 contributes to hepatic DTIC metabolism at higher substrate concentrations; and (c) CYP1A1 catalyzes extrahepatic metabolism of DTIC. PMID- 10473106 TI - Anticancer drug sensitivity and expression of multidrug resistance markers in early passage human sarcomas. AB - We have established new human sarcoma lines and examined their sensitivity to common antitumor drugs and expression of putative multidrug resistance (MDR) proteins. Eighty-two sarcoma samples were transplanted in nude mice. Fourteen of these sarcomas were established as tumor cell lines. We determined a chemosensitivity profile to antitumor drugs (MDR drugs = doxorubicin, mitoxantrone, and vincristine; non-MDR drugs = cisplatin, ifosfamide, and bleomycin) for each tumor line in vivo. Response to chemotherapy with doxorubicin and ifosfamide was observed in 30-50% of these tumor lines. Our results obtained with xenotransplants are similar to the results documented in clinical trials in which doxorubicin and ifosfamide are effective in 30-50% of the patients. Furthermore, we examined expression of MDR-relevant markers like P-glycoprotein, MDR-associated protein, lung resistance protein, and mdr1 mRNA in these xenotransplants. A relationship between mdr1 mRNA expression and response to doxorubicin was demonstrated in >90% of our tumor lines. In six sarcomas with mdr1 mRNA expression, five were resistant against doxorubicin and cross-resistant against several other drugs, whereas from eight sarcomas, which lacked detectable mdr1 mRNA, seven were sensitive to doxorubicin and other drugs. We found lung resistance protein or MDR-associated protein expressed in three resistant and mdr1 mRNA-positive sarcomas. These results demonstrate that mdr1 mRNA expression is a putative marker for drug resistance in our sarcoma lines. We conclude, therefore, that inherent P-glycoprotein expression might be also responsible for drug resistance occurring in treatment of patients with sarcomas. The established tumor lines are useful for additional investigations on mechanisms of drug resistance in sarcomas and as models for preclinical screening of new antitumor drugs. PMID- 10473107 TI - The Trk tyrosine kinase inhibitor CEP-701 (KT-5555) exhibits significant antitumor efficacy in preclinical xenograft models of human pancreatic ductal adenocarcinoma. AB - The aggressive behavior and poor prognosis of pancreatic ductal adenocarcinoma (PDAC) is associated with an increased expression of many growth factors and their cognate receptors. We have previously demonstrated the aberrant expression of the Trk receptors (Trks A, B, and C), enhanced tumor stromal expression of neurotrophins in primary PDAC specimens and human PDAC-derived cell lines, and a dose-dependent biological response of PDAC cells (in vitro invasiveness) to selective neurotrophins (Miknyoczki, S. J., et al., Int. J. Cancer, 81: 417-427, 1999). On the basis of these data, we have evaluated the therapeutic potential of inhibiting neurotrophin-Trk interactions using a selective and potent Trk tyrosine kinase inhibitor (CEP-701) in several preclinical models of human PDAC. CEP-701 is currently approved for clinical trials within the United States We demonstrate that CEP-701 administration at 10 mg/kg s.c. b.i.d. 5 days a week for 21-28 days inhibited tumor growth in a statistically significant manner in Panc 1, AsPc-1, BxPc-3, Colo 357, and MiaPaCa2 s.c. xenografts in athymic nude mice compared with vehicle-treated controls. Reductions in tumor growth volume of 50 70% relative to vehicle-treated controls were observed in xenografts responsive to CEP-701 administration. Significant reductions of in vivo PDAC tumor invasiveness were likewise observed in four of six CEP-701-treated rat tracheal xenografts implanted s.c. in athymic nude mice. The antitumor efficacy of CEP-701 was observed in the absence of pronounced morbidity or toxicity in vivo. Taken together, these data suggest that CEP-701 may be effective as a potential therapeutic agent in the treatment or management of PDAC. PMID- 10473108 TI - Paclitaxel and radiotherapy: sequence-dependent efficacy--a preclinical model. AB - The optimal sequence of a paclitaxel-radiation combination was investigated in vitro in two human colon adenocarcinoma cell lines, HT29 and LoVo. Three schedules of combined treatment were tested by clonogenic and flow cytometric assays. Paclitaxel was given 24 h prior to a single radiation shot (first schedule) or 24 h (second schedule) or 48 h (third schedule) before 3 days of concomitant radiation. Dose-response data were fit to a linear quadratic model, and mean inactivation dose and sensitizer enhanced ratio were calculated. In HT29 cells, the first and second schedule resulted in an additive effect, whereas a supraadditive interaction was observed with the third combination schedule. This effect was obtained with amounts of paclitaxel lower than IC50, which did not result in cell cycle perturbation, and with low radiation dose (2 Gy) that may be given in a clinical setting. LoVo cells were less sensitive to combined treatment than HT29 cells, switching from infraadditive (first and second schedule) to additive interaction (third schedule). Posttreatment recovery studies of third schedule showed a loss of cell survival in HT29 cells but not in LoVo cells. In contrast to LoVo cells, the third schedule in HT29 cells was able to induce perturbation of cell cycle kinetics, an effective impairment of DNA repair, and apoptotic cell death. HT29 and LoVo cells showed constitutional different characteristics: HT29 cells were more sensitive to paclitaxel exposure, less radiosensitive, and had a different cell cycle redistribution after radiation exposure than LoVo cells; moreover, HT29 cells showed a major propensity to undergo apoptosis. These results suggest that the radiosensitizing effect of paclitaxel was strictly schedule dependent, and the inhibition of DNA repair, cell cycle redistribution, and apoptosis could be the mechanisms for the induction of radiosensitization by paclitaxel. PMID- 10473109 TI - Lovastatin augments apoptosis induced by chemotherapeutic agents in colon cancer cells. AB - Beta-hydroxy-beta-methylglutaryl coA reductase inhibitors (HRIs) inhibit isoprenylation of several members of the Ras superfamily of proteins and therefore have important cellular effects, including the reduction of proliferation and increasing apoptosis. Significant toxicity at high doses has precluded the use of HRIs as a monotherapy for cancers. We therefore studied whether combinations of the HRI lovastatin with standard chemotherapeutic agents would augment apoptosis in colon cancer cells. In the colon cancer cell lines SW480, HCT116, LoVo, and HT29, lovastatin induced apoptosis with differing sensitivity. Pretreatment with lovastatin significantly increased apoptosis induced by 5-fluorouracil (5-FU) or cisplatin in all four cell lines. Lovastatin treatment resulted in decreased expression of the antiapoptotic protein bcl-2 and increased the expression of the proapoptotic protein bax. The addition of geranylgeranylpyrophospate (10 microM) prevented lovastatin-induced augmentation of 5-FU and cisplatin-induced apoptosis; mevalonate (100 microM) was partially effective, whereas cotreatment with farnesyl pyrophosphate (100 microM) had no effect. These data imply that lovastatin acts by inhibiting geranylgeranylation and not farnesylation of target protein(s). Our data suggest that lovastatin may potentially be combined with 5-FU or cisplatin as chemotherapy for colon cancers. PMID- 10473110 TI - Fenretinide activates caspases and induces apoptosis in gliomas. AB - The synthetic retinoid fenretinide (N-[4-hydroxyphenyl] retinamide or 4HPR) has been shown to not only inhibit cell growth but also to induce apoptosis in a variety of malignant cell lines. It is being tested presently for its potential as a chemopreventive agent against several cancers. A related retinoid, 13-cis retinoic acid (cRA), has been shown to have activity against gliomas in vitro as well as in a recent clinical study. The present study aimed at assessing the activity of fenretinide against glioma cells in vitro and comparing it with that of cRA at pharmacologically relevant doses. We hypothesized that the ability of fenretinide to induce apoptosis would make it more potent against gliomas than cRA. Four glioma cell lines (D54, U251, U87MG, and EFC-2) were treated with fenretinide (1-100 microM) and showed dose- and time-dependent induction of cell death. At pharmacologically relevant doses, fenretinide was more active against glioma cells than cRA because of its ability to induce apoptosis. Flow cytometric studies using D54 cells demonstrated no significant changes in the cell cycle distribution compared with untreated control, but a sub-G1 fraction consistent with apoptosis was detected. Terminal deoxynucleotidyl transferase-mediated nick end labeling assay indicated that the apoptotic fraction was cell cycle nonspecific. Fenretinide treatment resulted in cleavage of poly ADP-ribose polymerase, indicating an activation of the caspase 3. Immunofluorescence studies using the nuclear stain 4',6-diamidine-2'-phenylindole dihydrochloride showed nuclear condensation and an apoptotic morphology. Hence, this study demonstrates that, at clinically relevant doses, fenretinide is a potent inducer of apoptosis in gliomas acting via the caspase pathway. We also show that at clinically achievable doses, fenretinide has more activity against gliomas than comparable doses of cRA. The favorable side effect profile seen in previous clinical studies and the in vitro activity against gliomas demonstrated in this study suggest that fenretinide could be a promising therapeutic agent against gliomas. PMID- 10473111 TI - Identification of a MAGE-2-encoded human leukocyte antigen-A24-binding synthetic peptide that induces specific antitumor cytotoxic T lymphocytes. AB - Because MAGE-2 gene is expressed in a wide variety of malignant tumors and HLA A24 is the most common allele in the Japanese population and is also frequently present in Caucasians, the identification of MAGE-2-encoded peptide presented by HLA-A24 is, therefore, considered to be important in order to develop specific immunotherapy for malignant tumors using peptides as a vaccine. By using a MHC binding assay, eight peptides derived from MAGE-2 were found to bind with sufficient affinity to the HLA-A24 molecule. When the induction of specific cytotoxic T lymphocytes (CTLs) was examined using a simplified method, the highest human lymphocyte antigen (HLA) binder (EYLQLVFGI) in these peptides was able to elicit CTLs from unseparated peripheral blood mononuclear cells in HLA A24 healthy donors by stimulation with freshly isolated, peptide-pulsed peripheral blood mononuclear cells as antigen-presenting cells and also by using interleukin 7 and keyhole-limpet hemocyanin in a primary culture. The induced CTL could, thus, lyse HLA-A24 tumor cells expressing MAGE-2, as well as the peptide pulsed target cells, with antigen specificity in a HLA class I-restricted manner. The identification of this peptide may, thus, be of therapeutic value in peptide based vaccines for the treatment of several types of malignant tumors expressing MAGE-2. PMID- 10473113 TI - A role for phospholipase C-gamma-mediated signaling in tumor cell invasion. AB - The invasive and metastatic transformation of cancers often results in death. However, the mechanisms that promote this transformation remain unclear. Two closely related receptors, the epidermal growth factor receptor (EGFR) and ErbB2, are overexpressed in a significant percentage of breast and prostate carcinomas, among others, with this up-regulated signaling correlating with tumor progression. Previous studies in our laboratory have demonstrated that an EGFR phospholipase C (PLC)gamma-mediated motility-associated signaling pathway is rate limiting for tumor cell invasion in vitro and in vivo in one model of prostate carcinoma. Therefore, we investigated whether this PLCgamma signaling pathway also was rate-limiting for invasion in other tumor cell lines and types and whether this EGFR activity is subsumed by the closely related ErbB2. We determined the effects of PLCgamma signal abrogation by pharmacological (U73122) and molecular (expression of the dominant-negative PLCz) means on the in vitro invasiveness of tumor cells. Inhibition of PLCgamma signaling concomitantly decreased invasiveness of de novo-occurring transgenic adenocarcinoma mouse prostate (TRAMP) lines and the human breast cancer cell lines MDA-468 and MDA 231; these lines present up-regulated EGFR signaling. Because the prostate and breast cancer lines usually present autocrine stimulatory loops involving EGFR, we also examined transgenic adenocarcinoma mouse prostate C1 and MDA-468 treated with the EGFR-specific kinase inhibitor PD153035 to determine whether invasiveness is dependent on EGFR signaling. PD153035 reduced invasiveness to levels similar to those seen with U73122, suggesting that the autocrine EGFR stimulatory loop is functioning to promote invasiveness. To determine whether this signaling pathway also promotes invasiveness of ErbB2-overexpressing tumors, we examined the human breast carcinoma line MDA-361; again, U73122 inhibition of PLCgamma decreased invasiveness. In all situations, the inhibition of PLCgamma signaling did not decrease mitogenic signaling. Thus, the motility-associated PLCgamma signaling pathway is a generalizable rate-limiting step for tumor cell progression. PMID- 10473112 TI - Adenovirus-mediated E2F-1 gene transfer inhibits MDM2 expression and efficiently induces apoptosis in MDM2-overexpressing tumor cells. AB - The oncoprotein MDM2 binds and inactivates p53. MDM2 also binds to the tumor suppressor pRB, as well as E2F-1. E2F-1 is a transcription factor that regulates S phase entry and has been shown to cause apoptosis in some cell types when overexpressed. To investigate the effect of adenovirus-mediated E2F-1 overexpression, MDM2-overexpressing tumor cell lines were treated by mock infection, infection with an adenoviral vector expressing beta galactosidase, or E2F-1 (Ad5CMV-E2F-1). Western blot analysis confirmed significant overexpression of E2F-1 in Ad5CMV-E2F-1-infected cells. E2F-1 overexpression resulted in marked growth inhibition and rapid loss of cell viability. Ad5CMV-E2F-1 infection resulted in early S phase entry, followed by apoptotic cell death. E2F-1 overexpression was associated with a marked decrease in MDM2 levels and no evidence of increased Bax levels, whereas p53 and Bcl-2 levels remained undetectable. Cleavage of poly-ADP-ribose polymerase and caspase 3/CPP32 implicated activation of the caspase cascade in E2F-1-mediated apoptosis. These results indicate that adenovirus-mediated E2F-1 overexpression in MDM2 overexpressing tumor cells results in decreased MDM2 expression and widespread apoptosis. Because MDM2-overexpressing tumors are often resistant to p53 gene therapy, adenovirus-mediated E2F-1 gene therapy may be a promising alternative strategy. PMID- 10473114 TI - Surface membrane-expressed CD40 is present on tumor cells from squamous cell cancer of the head and neck in vitro and in vivo and regulates cell growth in tumor cell lines. AB - Because regional spread to lymph nodes without systemic spread is a relatively common event in squamous cell cancer of the head and neck (SCCHN), it is possible that lymphoid-related receptors or cytokines might directly impact the growth of these tumors. In the present study, we have shown by flow cytometry and Western blotting that the central lymphoid regulatory molecule, CD40, is expressed on the surface of all seven SCCHN tumor cell lines studied. Tumor cell lines also expressed epidermal growth factor (EGF) receptor, MHC class I, and CD95 (Fas) but did not uniformly express other important lymphoid regulatory molecules such as CD80, CD86, or interleukin (IL) 2 receptor components. CD40 ligation by trimeric CD40 ligand (CD40L) resulted in a 20-45% inhibition of tumor cell growth in three of seven cell lines tested. The cytokines IL-1alpha, IL-1beta, IL-2, IL-4, IL-6, IL-10, IL-11, and IL-15 neither inhibited nor stimulated growth in any of the cell lines tested. EGF had pleiotropic effects on cell growth; it inhibited growth in two cell lines, stimulated growth in one cell line, and had no effect in four cell lines. When coligation by EGF and CD40L was studied, additive or supra-additive growth inhibition was seen in four cell lines. Three cell lines were unaffected by EGF, CD40, or coligation with both reagents. Examination of tumor tissues from 12 previously untreated patients representing a broad spectrum of patients presenting with SCCHN demonstrated CD40 expression in all 12 tumor specimens. This study supports the notion that CD40 is a regulatory molecule for the growth of SCCHN. The important role of CD40-CD40L interactions in the regulation of immune cells in the lymph node and the unique high-level expression of CD40L by these immune cells lend support to the hypothesis that this ligand/receptor pair is an important mediator of cell growth in SCCHN. PMID- 10473115 TI - Osteopontin: possible role in prostate cancer progression. AB - Human prostate cancer has the propensity to metastasize to the bone where reciprocal cellular interactions between prostate cancer and bone cells are known to occur. Osteopontin (OPN), a noncollagenous bone extracellular matrix, is a secreted adhesive glycoprotein with a functional RGD cell-binding domain that interacts with the alpha(v)beta3 cell surface integrin heterodimer. OPN has been associated with malignant transformation as well as being ligand to the CD44 receptor. Polyclonal antibodies to human OPN (hOPN) were prepared, and specificity was shown by preabsorption with recombinant hOPN. The stimulatory effect of hOPN protein and the inhibitory effect of hOPN antibody on human prostate cancer cell lines LNCaP and C4-2 were assessed by induction or inhibition of anchorage-independent growth, respectively. Expression of hOPN mRNA in prostate cancer cell lines and human prostate cancer tissue specimens were measured by mRNA blot analysis. Protein expression was assessed by immunohistochemistry in human prostate cancer specimens and by Western blot analysis in prostate cancer cell lines. hOPN stimulated anchorage-independent growth of the human prostate cancer cell lines LNCaP and C4-2 in vitro. Antibodies to hOPN inhibited the growth-stimulatory effect by endogenous OPN, which can be overcome by the addition of exogenous hOPN. hOPN mRNA and protein are expressed in human prostate cancer cell lines in vitro and in clinical human prostate cancer specimens. These findings taken together suggest that OPN may act as a paracrine and autocrine mediator of prostate cancer growth and progression. PMID- 10473116 TI - Mesoderm-determining transcription in Drosophila is alleviated by mutations in TAF(II)60 and TAF(II)110. AB - In Drosophila, a coordinate interplay between the Rel transcription factor Dorsal and the basic Helix-Loop-Helix transcription factor Twist initiates mesoderm formation by activating the zygotic expression of mesoderm-determining genes. Here, we show that TBP-associated-factors (TAF(II)s) within the basal transcription factor TFIID mediate transcriptional activation by Dorsal and Twist. Dorsal interacts with TAF(II)110 and TAF(II)60, while Twist contacts TAF(II)110. The TAF(II):activator interactions mediate simple and synergistic transactivation by Dorsal and Twist in vitro. Mutations in TAF(II)60 or TAF(II)110 alleviate the transcription of Dorsal and Twist target genes. Gene dosage assays imply that an interplay of Dorsal and Twist with TAF(II)110 is critically required for the activation of mesoderm-determining gene expression in the Drosophila embryo. The results provide evidence that TAF(II)-subunits within the TFIID complex play an important role during the molecular events leading to initiation of mesoderm formation in Drosophila. PMID- 10473117 TI - Retinoic acid induces down-regulation of Wnt-3a, apoptosis and diversion of tail bud cells to a neural fate in the mouse embryo. AB - The tail bud comprises the caudal extremity of the vertebrate embryo, containing a pool of pluripotent mesenchymal stem cells that gives rise to almost all the tissues of the sacro-caudal region. Treatment of pregnant mice with 100 mg/kg all trans retinoic acid at 9.5 days post coitum induces severe truncation of the body axis, providing a model system for studying the mechanisms underlying development of caudal agenesis. In the present study, we find that retinoic acid treatment causes extensive apoptosis of tail bud cells 24 h after treatment. Once the apoptotic cells have been removed, the remaining mesenchymal cells differentiate into an extensive network of ectopic tubules, radially arranged around the notochord. These tubules express Pax-3 and Pax-6 in a regionally-restricted pattern that closely resembles expression in the definitive neural tube. Neurofilament-positive neurons subsequently grow out from the ectopic tubules. Thus, the tail bud cells remaining after retinoic acid-induced apoptosis appear to adopt a neural fate. Wnt-3a, a gene that has been shown to be essential for tail bud formation, is specifically down-regulated in the tail bud of retinoic acid-treated embryos, as early as 2 h after retinoic acid treatment and Wnt-3a transcripts become undetectable by 10 h. In contrast, Wnt-5a and RAR-gamma are still detectable in the tail bud at that time. Extensive cell death also occurs in the tail bud of embryos homozygous for the vestigial tail mutation, in which there is a marked reduction in Wnt-3a expression. These embryos go on to develop multiple neural tubes in their truncated caudal region. These results suggest that retinoic acid induces down-regulation of Wnt-3a which may play an important role in the pathogenesis of axial truncation, involving induction of widespread apoptosis, followed by an alteration of tail bud cell fate to form multiple ectopic neural tubes. PMID- 10473118 TI - Six6 (Optx2) is a novel murine Six3-related homeobox gene that demarcates the presumptive pituitary/hypothalamic axis and the ventral optic stalk. AB - We report on the isolation of a murine homeobox-containing gene, Six6 (Optx2), that shows extended identity in its coding region with Six3, the only member of the mammalian Six gene family known to be expressed in the optic primordium. Phylogenetic analysis demonstrates that Six6 and Six3 belong to a separate group of homeobox-genes that are closely related to the recently identified Drosophila optix. Earliest Six6 expression was detected in the floor of the diencephalic portion of the primitive forebrain, a region predicted to give rise to the neurohypophysis and to the hypothalamus. Later on, Six6 mRNA was found in the primordial tissues giving rise to the mature pituitary: the Rathke's pouch and the infundibular recess. In the optic primordium, Six6 demarcates the presumptive ventral optic stalk and the ventral portion of the future neural retina. In the developing eye. Six6 expression was detected in the neural retina, the optic chiasma and optic stalk, but not in the lens. When compared to Six6, Six3 expression pattern was highly similar, but with a generally broader transcripts distribution in the brain and in the visual system. We finally show that Six6 does not require Pax6 for its expression in the optic primordium, suggesting that Six6 acts on a parallel and/or independent pathway with Pax6 in the genetic cascade governing early development of the eye. PMID- 10473119 TI - Patterning signals acting in the spinal cord override the organizing activity of the isthmus. AB - The regionalization of the neural tube along the anteroposterior axis is established through the action of patterning signals from the endomesoderm including the organizer. These signals set up a pre-pattern which is subsequently refined through local patterning events. The midbrain-hindbrain junction, or isthmus, is endowed with such an organizing activity. It is able to induce graded expression of the Engrailed protein in the adjacent mesencephalon and rhombencephalon, and subsequently elicits the development of tectal and cerebellar structures. Ectopically grafted isthmus was also shown to induce Engrailed expression in diencephalon and otic and pre-otic rhombencephalon. Fgf8 is a signalling protein which is produced by the isthmus and which is able to mimic most isthmic properties. We show here that the isthmus, when transposed to the level of either rhombomere 8 or the spinal cord, loses its ability to induce Engrailed and cerebellar development in adjacent tissues. This is accompanied by the down-regulation of fgf8 expression in the grafted isthmus and by the up regulation of a marker of the recipient site, Hoxb-4. Moreover, these changes in gene activity in the transplant are followed by a transformation of the fate of the grafted cells which adjust to their novel environment. These results show that the fate of the isthmus is not determined at 10-somite stage and that the molecular loop of isthmic maintenance can be disrupted by exogenous signals. PMID- 10473120 TI - An octamer-binding site is crucial for the activity of an enhancer active at the embryonic met-/mesencephalic junction. AB - An enhancer sequence found in the Protease Nexin-1 (PN-1) gene was shown to drive lacZ expression specifically at the met-/mesencephalic junction in transgenic mouse embryos. A functional study of this enhancer has been performed to better understand the mechanisms regulating isthmic gene expression. An octamer-binding site for POU domain factors was found to be crucial for the activity of the enhancer in vivo. Comparative expression studies of POU domain factors, electrophoretic mobility shift assays and transient transfection experiments, strongly suggest that Brn-1/-2 regulate the enhancer activity in vivo. In addition, in vitro experiments indicated that FGF-8 was required for the maintenance of the enhancer activity, but not for the synthesis of Bn-1/-2. The data represents the first functional evidence for a role of POU factors in the regulation of met-/mesencephalic gene expression. It also implies that at least two regulatory pathways, namely the FGF-8 signaling and the octamer-binding site pathway, synergistically interact to control the PN-1 enhancer activity in vivo. PMID- 10473121 TI - Ubiquitous expression of a Drosophila adenomatous polyposis coli homolog and its localization in cortical actin caps. AB - We report the expression pattern of a new adenomatous polyposis coli (APC) homolog called E-APC during Drosophila development. E-APC protein is expressed in all embryonic and larval cells we have examined. In the early blastoderm embryo, we see a striking concentration of E-APC in the cortical actin caps. Microtubules are closely associated with these caps. Since human APC has been reported to bind to microtubules, we investigated whether the cortical E-APC co-localizes with tubulin. However, this was not the case, implying that the putative tubulin binding property of human APC is not well conserved. PMID- 10473122 TI - Xcat2 RNA is a translationally sequestered germ plasm component in Xenopus. AB - In Xenopus, the inheritance of germ plasm by a small subset of blastomeres during early development is thought to direct these cells into the germ cell lineage. We show that Xcat2 RNA, related to Drosophila nanos, is a germ plasm component that is translationally repressed during oogenesis. Xcat2 protein was not detected in oocytes at times prior to, or after its RNA was localized in germ plasm, suggesting Xcat2 RNA is functionally sequestered soon after transcription. Indeed, Xcat2 RNA is found in a dense non-polysomal compartment in oocytes. Repression of translation was not relieved by substituting the Xcat2 3'UTR with that of beta-globin. Immunodetection of Xcat2 protein during blastula and gastrula stages coincides with the time of symmetric segregation of the germ plasm and a net increase in the number of primordial germ cells. Xcat2 is capable of binding RNA in vitro and we propose that it may function to translationally regulate other RNAs specific to primordial germ cells. PMID- 10473123 TI - Translational control of nuclear lamin B1 mRNA during oogenesis and early development of Xenopus. AB - Cytoplasmic polyadenylation of specific mRNAs is commonly correlated with their translational activation during development. A canonical nuclear polyadenylation element AAUAAA (NPE) and cytoplasmic polyadenylation element(s) (CPE) are necessary and sufficient for polyadenylation during egg maturation. We have characterized cis-acting sequences of Xenopus nuclear lamin B1 mRNA that mediate translational regulation. By injection of synthetic RNAs into oocytes we show that the two CPE-like elements found in the 3'-untranslated region of B1 mRNA act as translational repressors in oocytes. The same CPEs in conjunction with the NPE confer transient polyadenylation and translational activation during egg maturation. Poly(A) length determination of the endogenous lamin B1 mRNA reveals a gradual increase of poly(A) tail length in early development up to mid blastula, and a shortening of poly(A) tails during gastrulation and neurulation. The same kinetic and extent of polyadenylation and poly(A) tail shortening is observed with synthetic RNAs injected into fertilized eggs. Polyadenylation and translational activation of these RNAs is independent of the two CPEs and a NPE during early development. While translational regulation of lamin B1 mRNA functions in parts via established mechanisms, the pattern of polyadenylation and deadenylation during early development points to a novel mode of translational regulation. PMID- 10473124 TI - Two distinct subgroups of Group B Sox genes for transcriptional activators and repressors: their expression during embryonic organogenesis of the chicken. AB - Group B Sox genes, Sox1, -2 and -3 are known to activate crystallin genes and to be involved in differentiation of lens and neural tissues. Screening of chicken genomic sequences for more Group B Sox genes identified two additional genes, Sox14 and Sox21. Proteins encoded by Sox14 and Sox21 genes are similar to each other but distinct from those coded by Sox1-3 (subgroup B1) except for the HMG domain and Group B homology immediately C-proximal of the HMG domain. C-terminal domains of SOX21 and SOX14 proteins function as strong and weak repression domains, respectively, when linked to the GAL4 DNA binding domain. These SOX proteins strongly (SOX21) or moderately (SOX14) inhibited activation of delta1 crystallin DC5 enhancer by SOX1 or SOX2, establishing that Sox14 and Sox21 are repressing subgroup (B2) of Group B Sox genes. This provides the first evidence for the occurrence of repressor SOX proteins. Activating (B1) and repressing (B2) subgroups of Group B Sox genes display interesting overlaps of expression domains in developing tissues (e.g. optic tectum, spinal cord, inner ear, alimentary tract, branchial arches). Within each subgroup, most expression domains of Sox1 and -3 are included in those of Sox2 (e.g. CNS, PNS, inner ear), while co expression of Sox14 and Sox21 occurs in highly restricted sites of the CNS, with the likely temporal order of Sox21 preceding Sox14 (e.g. interneurons of the spinal cord). These expression patterns suggest that target genes of Group B SOX proteins are finely regulated by the counterbalance of activating and repressing SOX proteins. PMID- 10473125 TI - Developmental expression analysis of murine autotaxin (ATX). AB - The murine homologue of the human motility-stimulating protein autotaxin (ATX) was identified as a BMP2 upregulated gene by subtractive cloning from mesenchymal progenitors C3H10T1/2 (Bachner, D., Ahrens, M., Betat, N., Schroder, D., Hoffmann. A., Lauber, J., Steinert, P., Flohe, L., Gross, G., 1998. Bmp-2 downstream targets in mesenchymal development identified by subtractive cloning from recombinant mesenchymal progenitors (C3H10T1/2). Dev. Dyn. 213, 398-411). ATX mRNA transcription is induced during BMP2 mediated osteo-/chondrogenic differentiation in vitro several orders of magnitude. To delineate a potential role for ATX in osteo-/chondrogenic development, its expression pattern during murine embryogenesis was examined in comparison with Col1a1 and Col2a1, a marker either of osteoblast, odontoblast and tendon or of chondrocyte development, respectively. Localization of murine ATX was first observed in the floor plate of the neural tube at day 9.5 of mouse embryonic development. Later, enhanced ATX expression levels were observed in proliferating subepithelial mesenchyme, during osteo-/chondrogenic and tooth development, in choroid plexus epithelium, in late kidney development, and in smooth muscles of the ductus deferens and the bladder. PMID- 10473126 TI - Migration of mesonephric cells into the mammalian gonad depends on Sry. AB - In mammals, the primary step in male sex determination is the initiation of testis development which depends on the expression of the Y-linked testis determining gene, Sry. The mechanisms by which Sry controls this process are unknown. Studies showed that cell migration from the adjacent mesonephros only occurs into XY gonads; however, it was not known whether this effect depended on Sry, another Y-linked gene, or the presence of one versus two X chromosomes. Here we provide genetic proof that Sry is the only Y-linked gene necessary for cell migration into the gonad. Cell migration from the mesonephros into the differentiating gonad is consistently associated with Sty's presence and with testis cord formation, suggesting that cell migration plays a critical role in the initiation of testis cord development. The induction of cell migration represents the earliest signaling pathway yet assigned to Sry. PMID- 10473127 TI - Expression pattern of the Tbr2 (Eomesodermin) gene during mouse and chick brain development. AB - The members of the T-box gene family share a highly conserved DNA binding domain named the T-domain, and important developmental functions. Here we report the cloning of chicken Tbr1 and of murine and chicken Tbr2 (orthologs of the Xenopus eomesodermin gene), the mapping of the murine Tbr2 to chromosome 9, and their pattern of expression during mouse and chick embryogenesis. Both Tbr 1 and 2 have a restricted and conserved domain of expression in the telencephalic pallium of the two species. Chick Tbr2 has a specific and dynamic expression in the gastrulating embryo. PMID- 10473128 TI - Xenopus elav-like genes are differentially expressed during neurogenesis. AB - In Xenopus, three neural-specific elav-like genes (ELGs) have been identified, elrB/Xel-1, elrC and elrD. With the aim to highlight possible differences in the regulation of these genes, we compared their expression patterns during development. We had previously shown that elrB is expressed from the early tailbud stage onwards, in both the central and peripheral nervous system. Here we show that both elrC and elrD are expressed earlier than elrB in the developing neural tube and the cranial ganglia, with different temporal specificities. Double in situ hybridizations on brain cross-sections allowed us to define precisely the expression domains of elrB, elrC and elrD in the brain at the tailbud stage. What emerges from this study is a differential distribution of ELGs transcripts in the hindbrain. Also, double labeling with a motor neuron marker shows that in stage 41 tailbud embryos, elrD remains strongly expressed in motor neurons whereas elrC is mostly expressed in non-motor neuron cells. PMID- 10473129 TI - Sagittal band expression of COUP-TF2 gene in the developing cerebellum. AB - In the developing cerebellum, the medio-lateral compartmentalization of the adult cerebellum is preceded by the transient expression of factors which divide the cortex into similar parasagittal stripes. Here we report that COUP-TF2, an orphan member of the nuclear receptor family which suppresses RA actions by forming heterodimers with RXR, shows a pattern of sagittal bands in developing mouse cerebellum. The band pattern changes according to the developmental stage. At embryonic day 13 it is expressed in the lateral half of the cerebellum, but at later stages the expression is divided into several parasagittal bands. By postnatal day 5 the COUP-TF2 expression substantially decreases to low, but detectable, levels. PMID- 10473130 TI - Dynamic expression of TSC-22 at sites of epithelial-mesenchymal interactions during mouse development. AB - The leucine zipper transcription factor TSC-22 (TGF-beta1 Stimulated Clone-22) was first isolated from a mouse osteoblast cell line as an immediate-early target gene of TGF-beta1. However, work with other cell lines, as well as with a Drosophila homolog, bunched, suggests that it is an effector gene of various growth factors and potentially involved in the integration of multiple extracellular signals. Throughout mouse embryogenesis TSC-22 is expressed in a dynamic pattern. Although early TSC-22 expression is ubiquitous in 6.5 day embryos, as development proceeds TSC-22 expression is upregulated at sites of epithelial-mesenchymal interactions such as the limb bud, tooth primordiurn, hair follicle, kidney, lung, and pancreas. TSC-22 is also expressed in many neural crest-derived tissues including the mesenchyme of the branchial arches, the cranial, dorsal root, and sympathetic ganglia, as well as the facial cartilage and bone. Other areas of expression are the otic and optic vesicles, the heart, and cartilage and bone forming regions throughout the embryo. PMID- 10473131 TI - Expression pattern of macrophage migration inhibitory factor during embryogenesis. AB - Although macrophage migration inhibitory factor (MIF) was originally identified as a lymphokine that inhibits the migration of macrophages, its ubiquitous expression suggests it may have a role beyond the immune system. Here we report a detailed characterization of MIF expression during mouse embryogenesis. The MIF expression pattern was found to parallel tissues specification and organogenesis. PMID- 10473132 TI - Cloning and expression analysis of a mouse gene related to Drosophila odd skipped. AB - The Drosophila pair-rule gene odd-skipped (odd) and two related genes, sister of odd (sob) and bowel (bowl), encode zinc finger containing proteins, two of which play important roles in embryonic development probably functioning as transcription factors. Here we report the cloning and expression analysis of a mouse gene related to odd, odd-skipped related 1 (Osr1). During early embryogenesis Osr1 is expressed in the intermediate mesoderm and in a dynamic pattern during limb and branchial arch development. PMID- 10473133 TI - Dynamic expression of ornithine decarboxylase in hair growth. AB - Ornithine decarboxylase (ODC) is the key enzyme in the synthesis of polyamines, small cationic molecules believed to have a role in many cellular processes such as cell migration, proliferation and differentiation. We show that ODC expression is associated with cell proliferation and commitment in hair follicle development and hair growth. In embryonic epidermis, ODC is expressed in ectodermal cells at sites where follicles develop, and persists in cells at the leading edge of the follicle placode. ODC is abundantly expressed in proliferating bulb cells of anagen follicles, except for a pocket of cells at the base of the bulb. Entry of the follicle into catagen is accompanied by a down-regulation of ODC expression, which is not resumed until a new follicle is initiated. In vibrissae, ODC expression is more complex. ODC is expressed not only in the bulb but also in the hair shaft, presenting a striking biphasic pattern. Additionally, ODC is expressed in a group of outer root sheath cells in the vicinity of the follicle bulge, the putative site of hair follicle stem cells. PMID- 10473134 TI - Expression of the mouse Delta1 gene during organogenesis and fetal development. AB - Cell-to-cell communication mediated by the evolutionary conserved Notch signalling pathway regulates cell fate decisions and patterning in various tissues in diverse organisms (Artavanis-Tsakonas et al., 1995, Science 268, 225 232). Signalling between neighboring cells is transduced by binding of DSL and Notch proteins which interact as ligand (DSL) and receptor (Notch). Mouse Delta1 (delta-like 1; Dll1) encodes one of the four known mammalian DSL proteins and is essential for normal somitogenesis and neuronal differentiation. Here, we describe Delta1 expression during organogenesis and fetal development using the highly sensitive histochemical detection of the lacZ gene product expressed from a targeted Delta1:lacZ knock-in allele (Dll1(lacZ)). We find that Delta1 is expressed in epithelial ducts of several organs, skeletal and smooth muscles, the central nervous system, as well as some sensory epithelia. PMID- 10473135 TI - Expression of Delta1 and Serrate1 (Jagged1) in the mouse inner ear. AB - The Notch signalling pathway is thought to play a key part in controlling the production of sensory hair cells in the vertebrate inner ear via lateral inhibition; but there is disagreement as to which Notch ligands are expressed in hair cells as they develop. We show, using a mouse Delta1:LacZ knock-in as a reporter, that nascent hair cells, but not their neighbours, express Delta1. Expression of Serrate1 (Jagged1), meanwhile becomes restricted to the supporting cells of each sensory patch. Delta1 is also expressed: (a) at early stages, at the site of otic neurogenesis; and (b) in scattered cells of the endolymphatic sac, as is Serrate1. PMID- 10473136 TI - Expression of GATA-2 in the developing avian rhombencephalon. AB - Here we describe the expression pattern of GATA-2 in the developing chick hindbrain. We found that during the early period of neurogenesis this gene specifically labels a particular neuronal subtype, the contralateral vestibular acoustic neurons of rhombomere 4, and that expression of this gene in these cells is transient. We also found that GATA-2 labels a broader territory of the ventral neural tube at later stages. As well as this hindbrain expression, we also describe expression of GATA-2 in the otic vesicle, oculomotor nucleus, third nerve and metanephros. PMID- 10473137 TI - Expression of the chicken angiotensin II receptor: atypical pattern compared to its mammalian homologues. AB - We have recently cloned and characterized pharmacologically a chicken angiotensin II receptor (cAT). To evaluate its putative role in developmental processes, we investigated its spatio-temporal distribution in the chicken embryo up to E14. The cAT mRNA is expressed in a developmental manner in the mesonephros and allantois, as well as in the heart, branchial arches or limbs. These results, the first to report the embryonic distribution of an angiotensin receptor in a non mammalian species, show that its expression pattern does not correspond to either one of the two angiotensin receptor types in mammalian species. PMID- 10473138 TI - Lbx2, a novel murine homeobox gene related to the Drosophila ladybird genes is expressed in the developing urogenital system, eye and brain. AB - We describe the cloning, expression pattern, and genomic organization of Lbx2, a murine homologue of the Drosophila and mammalian ladybird genes. Lbx2 includes a homeodomain motif most closely related to those of Lbx1 and the Drosophila ladybird proteins. Lbx2 transcripts are first detected at E10.5 when they are located in the gonadal component of the urogenital ridge. Expression of Lbx2 dramatically increases by E11.5 in the urogenital ridges, and in the cranial surface ectoderm. At later stages, Lbx2 transcripts are expressed in the brain and organs derived from the urogenital ridge, including the gonadal tubercle, kidneys, and adrenal glands. From E14.5 to birth, Lbx2 expression is evident in the developing retinal neuroepithelium and the vibrissa. PMID- 10473139 TI - Kidney-specific cadherin (cdh16) is expressed in embryonic kidney, lung, and sex ducts. AB - Cdh16 was initially described as a truncated cadherin expressed in the adult rabbit kidney. We have analyzed the expression pattern of cdh-16 during mouse embryogenesis, and show that cdh-16 transcripts are present in ureter-derived epithelia of the metanephric kidney. In addition, we demonstrate that cdh-16 is also transiently expressed in the epithelia of embryonic sex ducts and the lung of the embryo. PMID- 10473140 TI - Developmentally regulated expression of mtprd, the murine ortholog of tprd, a gene from the Down syndrome chromosomal region 1. AB - The gene tprd, which contains three tetratricopeptide domains, has been recently localized in the Down syndrome (DS) chromosomal region 1. We have cloned a cDNA encoding part of the murine ortholog of tprd and used it to characterize the expression pattern of this gene during development and at the adult stage. At E8.5 the expression is uniform. In the later stages of embryogenesis, although expression remains ubiquitous, a pattern of tissues with particularly high expression develops: the strong expression is restricted to non proliferating zones of the nervous system such as the external layer of the cortex, the spinal cord, the cranial and root ganglia and the nerves. In the brain of adult mouse the strongest signals are observed in layers II-III and V-VI of the cortex, in the hippocampus and in the cerebellum, which correspond to the abnormal brain regions seen in DS patients. PMID- 10473141 TI - Expression of three Rx homeobox genes in embryonic and adult zebrafish. AB - The paired-class homeobox gene, Rx, is important in eye development. In this study we analyze expression patterns of three zebrafish Rx genes (Zrx1, 2, 3) in embryos and adults. All three genes show dynamic spatiotemporal patterns of expression. Zrx3 is expressed earliest, in the anteriormost region of the neural plate, in regions that give rise to ventral diencephalon and retinae. As development proceeds, Zrx3 expression is reduced in the lateral optic primordia, and is absent in the optic cup, but is retained at the ventral midline of the diencephalon, and is expressed in hypothalamus in the adult. As the neural retina begins to differentiate, Zrx3 is re-expressed in a subset of cells in the inner nuclear layer, presumably bipolar cells, and this expression is retained in the adult. In contrast, Zrx1/2 have a slightly later onset of expression, are initially coincident with Zrx3, but then become complementary, remaining on in the optic primordia but disappearing from the ventral midline of the diencephalon. Zrx1/2 are down-regulated as the retina differentiates, except in the outer nuclear layer where they continue to be expressed at high levels in cone, but not rod, photoreceptors. This is the first transcription factor described that distinguishes between cone and rod photoreceptors. PMID- 10473142 TI - Amino acid sequence and embryonic expression of msr/apj, the mouse homolog of Xenopus X-msr and human APJ. AB - We have recently identified a new G protein-coupled receptor, X-msr, whose expression is associated with the endothelial lineage in Xenopus laevis (Devic, E., Paquereau, L., Vernier, P., Knibiehler, B., Audigier, Y., 1996. Expression of a new G protein-coupled receptor X-msr is associated with an endothelial lineage in Xenopus laevis. Mech. Dev. 59, 129-140). Based on its structural analogy to the human orphan receptor APJ, we cloned the murine msr/apj receptor and analyzed its expression in developing tissues. As observed for X-msr, msr/apj transcripts are detected in the endothelium of the primary blood vessels and the forming heart. In addition, they are expressed in somites, limb bud and branchial arches. This expression pattern is distinct from that of the Flk1 gene and suggests that the msr/apj gene is expressed in a subpopulation of endothelial precursors and a mesenchymal population derived from paraaxial mesoderm. PMID- 10473143 TI - Dupuytren's disease. The way forward? AB - Histologically, Dupuytren's disease has been compared to the process of neoplasia because of fibroblast proliferation, recurrence, chromosomal abnormalities and antigenic profiles. However, a comparison of Dupuytren's tissue with the granulation tissue formed in wound healing could be more valid. Histology reveals similarities in cell types, proliferation, vascularity and collagen morphology. Pharmacologically, both tissues have a similar range of agonist and antagonist responses. Biochemical analysis reveals new collagen synthesis, an increased ratio of type III to type I collagen, and similar changes of the ground substance in both processes. Considering such similarities perhaps it is possible to regard some of the models used for the investigation of wound healing and granulation tissue as the missing experimental "model" for the study of Dupuytren's disease. Recently great strides have been made in the basic understanding of wound biology, and such a comparison might well provide novel therapeutic options for Dupuytren's disease. PMID- 10473144 TI - The Simmen classification of wrist destruction in rheumatoid arthritis. Experience in patients with early disease. AB - In this radiological study, bilateral radiographs of the wrist in 48 patients with early rheumatoid arthritis (RA) were graded by the Simmen classification, at different times, to determine its potential use in early disease. Interobserver agreement was good. The Simmen classification of wrist destruction in RA could be applied to the radiographs of approximately 50% of patients with early disease. It has some consistency over time. PMID- 10473146 TI - Total wrist arthrodesis using bowed crossed K-wires. AB - A method of total wrist arthrodesis using a combination of autogenous iliac crest bone graft and "bowed" crossed Kirschner wires is described. The method of bowing the K-wires results in a compressive force on the iliac bone graft. This technique resulted in bony union of 22 wrists in 20 patients. The mean time to union was 12 weeks (range, 8-14 weeks). There were no major postoperative complications. The advantages of this technique are its simplicity, versatility, and reliability which mean that special internal fixation devices are not needed. PMID- 10473145 TI - Synovectomy combined with the Sauve-Kapandji procedure for the rheumatoid wrist. AB - The aim of synovectomy-stabilization (synovectomy combined with the Sauve Kapandji procedure) of the rheumatoid wrist is to obtain a stable painless wrist, retaining enough mobility for function. Thirty-nine wrists were retrospectively examined, at a mean follow up of 64.8 months. The improvement in pain was very significant. We noticed a decrease in wrist motion affecting both flexion and radial deviation. The arthritic change in the wrist continued to increase. We noticed a mean ulnar shift of 2.2 mm and a mean increase in the radial deviation of the wrist of 7 degrees. Only transfer of the extensor carpi radialis longus tendon to the extensor carpi radialis brevis tendon was effective in correcting radial deviation of the carpus. Ninety-seven per cent of patients were very satisfied or satisfied. These encouraging results, even at advanced stages of wrist arthritis, have prompted us to lessen the indications for wrist arthrodesis. PMID- 10473147 TI - Pseudarthrosis of the middle metacarpal following intramedullary fixation of a wrist arthrodesis. AB - We describe the case of a 36-year-old woman who presented with an unusual seronegative monoarthropathy of the right wrist and who was treated with fusion by the method of Clayton (1965). The patient probably sustained a metacarpal fracture during intramedullary Steinmann pin insertion, and subsequently developed a pseudarthrosis. PMID- 10473148 TI - A comparison of dominant and non-dominant hand strengths. AB - This study compares dominant and non-dominant hand strength in both right- and left-handed participants. Maximum voluntary contraction (MVC) of the first dorsal interosseous (FDI) muscle, power grip strength and pulp-to-pulp pinch strength were assessed under carefully controlled conditions. No significant differences were observed between dominant and non-dominant hands in left-handed participants for all tests. Small but significant differences (0.1-3%) were observed between dominant and non-dominant hands in right-handed participants for all three tests. These differences were much smaller than those reported in other studies. Furthermore, considerable variability was observed in the relative strengths of the two hands for each participant. We conclude that clinicians must be cautious when using the '10% rule' to make comparisons between injured and uninjured hands. PMID- 10473149 TI - The detection of feigned hand weakness using the five position grip strength test. AB - This study assessed the sensitivity and specificity of the five-position grip strength test (5PGST) for detecting feigned hand weakness in healthy volunteers. The 5PGST reliably detected feigned hand weakness in only 15% of cases. Thirty three per cent of volunteers demonstrated an entirely normal pattern of grip strength when feigning weakness. PMID- 10473150 TI - Finger dominance. AB - Two hundred and ninety-eight normal participants were reviewed to assess finger dominance. Participants were recruited from several large hospitals across the north-west of England and the study was done under blind conditions. From the 298 people originally in the trial, six had to be excluded as the dominant finger could not be confirmed with certainty. The index finger was the most dominant in 82.5% of the test group. However, in 10.6% of the test group, the middle finger was found to be the most dominant. PMID- 10473151 TI - The heterodigital reversed flow neurovascular island flap for fingertip injuries. AB - We report the results of pulp reconstruction with a new heterodigital reverse flow island flap. A dorsolateral flap from the middle phalanx, based on the digital artery is raised from the adjacent uninjured finger. The common digital artery, between the injured finger and the donor finger, is ligated and transected just before its bifurcation. The two converging branches of the digital arteries can be entirely mobilized as a continuous vascular pedicle for the flap. Thus the vascularization is now supplied by reverse flow through the proximal transverse digital palmar arch of the injured finger. To provide sensation the dorsal branch of the proper digital nerve from the donor finger can be included in the flap. Six reverse heterodigital island flaps were used in patients. In five patients the flap was used for pulp reconstruction and in one case for covering a dorsal digital defect. In one case mild venous congestion occurred. Good skin coverage with supple and well-vascularized skin was obtained in each patient. The static two-point discrimination over the flap was between 6 and 15 mm. This new procedure is indicated for extensive pulp defects in fingers in which reconstruction cannot be done using other flaps and as an alternative to microsurgical reconstruction. PMID- 10473152 TI - Distal thumb reconstruction using a mini wrap-around flap from the great toe. AB - We have used a great toe mini wrap-around flap for reconstruction of the thumb at, or distal to, the interphalangeal joint. Our series included 12 patients with traumatic amputations. A flap including the entire nail and most of the distal phalanx of the great toe was used. Eleven of the grafts survived. Sensibility was good with an average of 10 mm static two-point discrimination (range, 5-15) and there were no complaints of cold intolerance. All patients were pleased with the appearance of the thumb and there was no significant morbidity at the great toe donor site. The great toe mini wrap-around flap is an excellent reconstruction technique for selected patients with distal thumb amputations. PMID- 10473153 TI - Primary reconstruction of a degloved middle finger with a temporoparietal free flap. AB - The treatment of complete ring avulsion injuries is very demanding, and frequently the results are less than optimal. We present a case of a degloved middle finger reconstructed with a temporoparietal fascial free flap and skin grafting. This produced an aesthetic and functional finger. PMID- 10473154 TI - An anterior portal for wrist arthroscopy. Anatomical study and case reports. AB - We have found that the standard portals described limit arthroscopic access to and visualization of some areas of the radiocarpal joint. We describe a radial anterior portal, which we believe is useful in the arthroscopic treatment of wrist conditions and describe its use in clinical practice. PMID- 10473155 TI - Resection of the distal scaphoid for scaphotrapeziotrapezoid osteoarthritis. AB - Twenty-one patients with symptomatic scaphotrapeziotrapezoid osteoarthritis were treated with partial distal scaphoid excision. In 12 wrists the joint defect was filled with either capsular or tendinous tissue, while in nine no fibrous interposition was done. At an average follow-up time of 29 (range, 12-61) months, 13 wrists were painfree, while eight had occasional mild discomfort. Mean wrist flexion-extension was 119 degrees. Grip and pinch strength improved by an average of 26% and 40% respectively compared with their preoperative status. Fifteen patients returned to their original jobs, while six, who were unemployed, felt unrestricted for activities of daily living. Although patient satisfaction was comparable for both types of treatment, the wrists without fibrous interposition showed significantly greater wrist flexion-extension than patients with soft tissue interposition. Removal of the distal scaphoid resulted in a DISI pattern of carpal malalignment in 12 wrists. At follow-up, none of these wrists showed further joint deterioration due to residual malalignment. PMID- 10473156 TI - A patient-reported comparison of trapeziectomy with Swanson Silastic implant or sling ligament reconstruction. AB - A retrospective review of two types of operations for carpometacarpal osteoarthritis of the thumb was done for patients operated on between 1991 and 1996. Follow-up ranged from 18 to 90 months (mean 62 months). Fifty-eight Swanson Silastic arthroplasties and 56 sling excision arthoplasties were reviewed. Eight patients with Swanson arthroplasties underwent removal of the implant. Eight patients in the sling excision group required further surgery. These patients were excluded from further analysis. Questionnaires about pain, general satisfaction and function were sent to the other patients and 87 responses were received (sling 45, implant 42). In the implant group significantly better results were obtained for pain at 1 year, carrying a milk bottle and taking a handbrake off a car, and overall function. We conclude that trapeziectomy combined with Swanson implant gives better results in the short term if there are no complications of the operation. PMID- 10473157 TI - The sex ratio and rate of reoperation for Dupuytren's contracture in men and women. AB - We identified all patients treated by local fasciectomy at the Department of Hand Surgery, Uppsala University Hospital between 1965 and 1996. A total of 2375 operations were performed on 1600 patients. We found a male:female ratio of 5.9:1. Women had a higher mean age at first operation (62.4 years) than men (59.8 years). One-third of the men required repeated surgery and one-quarter of the women. Early age at first operation was associated with recurrent disease. PMID- 10473158 TI - Clinicopathological characteristics of melanomas of the hand. AB - A 25-year clinicopathological review of 19 patients with malignant melanoma of the hand is reported. There were 11 women (median age 73 years) and eight men (median age 70 years). Eleven tumours were subungual, six on the dorsum of the hand, and two had a palmar location. Seven tumours were acral lentiginous melanomas, six were superficial spreading melanomas, and five were of the nodular subtype. Eleven patients presented with localized disease (clinical stage I), four with local spread (clinical stage II), and four with regional lymph node metastases (clinical stage III). Nine of the patients have died of the disease, with a median survival time of 30 months (range, 10-84 months). The subungual melanomas were thicker and the patients were older at presentation compared with the non-subungual sites. Review of our patients emphasized the need for early diagnosis. PMID- 10473159 TI - Failed endoscopic carpal tunnel release. Operative findings and results of open revision surgery. AB - From 1994 to 1997, 22 patients (24 wrists) underwent open revision carpal tunnel release for persistent carpal tunnel syndrome after a primary endoscopic release. The age range was from 21 to 77 years. At the time of revision surgery, 22 wrists had an incomplete release of the flexor retinaculum and two patients had median nerve transection (one partial and one complete). One patient had release of Guyon's canal and not the carpal tunnel. After the open revision carpal tunnel release, 20 patients returned to work with five patients returning to jobs of lighter duty. In addition, these 20 patients had significant improvement in symptoms. The remaining two patients had sustained a median nerve injury and did not return to work. One of these patients developed a painful neuroma in continuity of the median nerve which required vein wrapping with a saphenous vein graft. This study indicates that endoscopic release of the flexor retinaculum holds the same risks and complications as open release. Based on our study we believe that patients with persistent carpal tunnel syndrome after failed endoscopic flexor retinaculum release can be successfully treated with open release. PMID- 10473160 TI - Palmar capsulodesis for treatment of symptomatic hyperextensibility of the metacarpophalangeal joints. AB - A modification of the technique of palmar capsulodesis described by Filler et al. (1976) for the treatment of hyperextensibility in the metacarpophalangeal joint of the thumb was used in the metacarpophalangeal joints of the fingers. Seven of eight patients regained normal range of motion without pain. PMID- 10473161 TI - Double second toe transfer in congenital hand anomalies. AB - A series of 14 patients with congenital hand anomalies who received staged double second toe transfers to the same hand for restoration of function or form were reviewed retrospectively. There were three children with constriction ring syndrome, two with symbrachydactyly and nine with transverse absence (failure of formation). There were different indications, technical difficulties and results with the various anomalies. All transferred toes were mobile and sensate, and were reported to be of benefit in both function and appearance. However, secondary surgical procedures were required in all patients. PMID- 10473162 TI - Intramedullary fixation by resorbable rods in a comminuted phalangeal fracture model. A biomechanical study. AB - The mechanical rigidity of three different methods of resorbable intramedullary fixation (bone peg, and polyglycolide rods with and without interlocking) was assessed in a comminuted phalangeal fracture model and the results compared with two commonly used internal fixation devices (lateral plate, crossed K-wires) in a cadaver model. Each fixation technique was tested for its biomechanical strength in apex palmar bending, compression and torsion. Failure testing for the three resorbable methods was also done. The results showed that lateral plating provided the best rigidity in apex palmar bending and torsion, followed by intramedullary bone peg fixation. All resorbable intramedullary fixations had rigidity that was at least the same as crossed K-wires. For the torque test, polyglycolide rods with interlocking provided better rigidity than without interlocking. There was no significant difference between the different methods in the compression test, except that the intramedullary bone peg was significantly stiffer than K-wires. PMID- 10473163 TI - A sporadic case of tetramelic mirror-image polydactyly and unilateral tibial hypoplasia without associated anomalies. AB - We report a sporadic case of tetramelic mirror-image polydactyly in combination with unilateral tibial aplasia. No chromosomal or other associated abnormalities were found. The appearance and function of the upper and lower limbs were improved by surgery. PMID- 10473164 TI - Intraosseous ganglion of the trapezium in communication with the flexor carpi radialis tendon sheath. AB - We report a case of an intraosseous ganglion of the trapezium that communicated with the flexor carpi radialis tendon sheath. The findings support the hypothesis that intraosseous ganglia arise from penetration of bone by synovial tissue or fluid. PMID- 10473165 TI - Advanced carpal collapse associated with congenital hypoplastic thumb. AB - We present the long term sequelae of two patients with congenital hypoplastic thumbs. Neither had surgical correction in childhood, but maintained adequate function in their hands. However, both have developed progressive carpal collapse in adulthood, leading to painful wrists. PMID- 10473166 TI - Palmar lunate trans-scaphoid, trans-triquetral fracture-dislocation. AB - A palmar lunate dislocation associated with fractures of the scaphoid and triquetrum and an avulsion fracture of the radial styloid is described. The injury resulted from a high energy dorsiflexion/axial compression force. Open reduction of the dislocated lunate and internal fixation of the scaphoid were done as an emergency procedure. A second operation was required to internally fix the displaced triquetral fracture and reduce the lunotriquetral dissociation. A good result was documented at 6 months. PMID- 10473167 TI - Assessment of vascularized fibular graft one year after reconstruction of the wrist after excision of a giant-cell tumour. AB - We report a patient in whom the distal radius was resected for a giant cell tumour and the bone defect was replaced using a vascularized proximal fibular graft. The graft was viable and hypertrophied and normal callus formed on the distal radius. Due to chronic instability of the wrist the patient underwent revision arthrodesis 1 year after resection. Microscopic studies of the epishyseal region of the fibula showed wide necrosis of the graft with active creeping substitution. Despite the good technical result of the vascularized fibular graft, the vascularization was incomplete in the proximal epiphysis. We discuss possible reasons for this. PMID- 10473168 TI - Recurrent osteoblastoma of the hamate bone. A two-stage reconstruction with a free vascularized iliac crest flap. AB - An osteoblastoma in a carpal bone is very rare and presents a problem of reconstruction after wide tumour excision. We report a case of recurrent osteoblastoma of the right hamate bone with involvement of the ulnar carpal bones and soft tissues that was successfully treated by en bloc resection, temporary interposition of bone cement and fixation with K-wires, followed by reconstruction with a free vascularized iliac crest flap, tailored to the exact size of the defect, in a second procedure. Rapid fusion was achieved and hand function preserved with no evidence of recurrence 3 years postoperatively. PMID- 10473169 TI - Effects of veratrine and paeoniflorin on the isolated rat aorta. AB - The interactions and mechanisms between veratrine and paeoniflorin on the isolated rat aorta were studied. Veratrine (1x10(-6) to 1x10(-4) g/ml) could induce contraction on the isolated rat aorta in a concentration-related manner. Paeoniflorin had no effect on the isolated rat aorta. Pretreatment with prazosin (1x10(-6) M) and nifedipine (1x10(-6) M) but not yohimbine (1x10(-5) M) could decrease the tension of contraction induced by veratrine (1x10(-4) g/ml). Sodium nitroprusside (1x10(-4) M) could inhibit the contraction induced by veratrine (1x10(-4) g/ml) with or without endothelium, whereas methylene blue (5x10(-5) M) could increase the contraction induced by veratrine (1x10(-4) g/ml). Treatment with veratrine (1x10(-4) g/ml) could decrease the tension of contraction induced by norepinephrine (1x10(-6) M) or phenylephrine (1x10(-4) M). The inhibition of veratrine on norepinephrine-induced contraction was potentiated by L-arginine (1x10(-4) M) and reversed by L-NAME (1x10(-5) M). Paeoniflorin (1x10(-4) M) could decrease the tension of contraction induced by veratrine (1x10(-4) g/ml) and methylene blue (5x10(-5) M). The inhibition of paeoniflorin on veratrine was more potent on rat isolated aorta with endothelium than without endothelium. Ryanodine (1x10(-5) M) and Ca2+ -free medium could inhibit methylene blue-induced contraction. From the above results, the relaxation of veratrine on the norepinephrine-induced contraction might be related to the increase of NO and cGMP. The contraction of veratrine on the isolated rat aorta was via the increase of intracellular calcium which was inhibited by paeoniflorin. PMID- 10473170 TI - Pharmacological effects of Agapanthus africanus on the isolated rat uterus. AB - The Agapanthus africanus plant is used by South African traditional healers as a phytomedicine in herbal remedies to treat pregnancy-related ailments and to augment labour. It has already been shown that an aqueous extract of A. africanus causes smooth muscle contractions in the isolated uterus and ileum preparations. In the present study, the effects of an aqueous extract of A. africanus leaves was examined on receptor systems involved in contraction of the uterine smooth muscle in order to determine the mechanism of its pharmacological effect relevant to its ethnic use to augment labour. The extract was tested on the isolated rat uterus preparation. The aqueous extract of A. africanus leaves was found to exhibit agonist activity on uterine muscarinic receptors and to promote the synthesis of prostaglandins in the oestrogenized rat uterus. Some pharmacological justification for the ethnic use of A. africanus as a herbal oxytocic in prolonged labour has been provided. PMID- 10473171 TI - Ex vivo and in vivo investigations of picroliv from Picrorhiza kurroa in an alcohol intoxication model in rats. AB - Picroliv, the active constituent isolated from the plant Picrorhiza kurroa, was evaluated as a hepatoprotective agent against ethanol-induced hepatic injury in rats. Alcohol feeding (3.75 g/kg x45 days) produced 20-114% alteration in selected serum (AST, ALT and ALP) and liver markers (lipid, glycogen and protein). Further, it reduced the viability (44-48%) of isolated hepatocytes (ex vivo) as assessed by Trypan blue exclusion and rate of oxygen uptake. Its effect was also seen on specific alcohol-metabolizing enzymes (aldehyde dehydrogenase, 41%; acetaldehyde dehydrogenase, 52%) in rat hepatocytes. The levels of these enzymes were found to be reduced in the cells following alcohol intoxication. Ethyl alcohol also produced cholestasis (41-53%), as indicated by reduction in bile volume, bile salts and bile acids. Picroliv treatment (3-12 mg/kg p.o. x45 days) restored the altered parameters in a dose-dependent manner (36-100%). PMID- 10473172 TI - Acute toxicity and general pharmacological effect on central nervous system of the crude rhizome extract of Kyllinga brevifolia Rottb. AB - Acute toxicity and general pharmacological activities of the crude hydro alcoholic rhizome extract of Kyllinga brevifolia Rottb., a popular medicine used in Paraguay, were investigated on mice. The intraperitoneal LD50 was found to be 575 mg/kg. Oral administration of doses up to 3.0 g/kg did not provoke any toxic symptoms. Oral administration of 100 mg/kg of the extract induced a significant increase in gastrointestinal transit. In open field studies, a decrease of spontaneous locomotor activity, piloerection, passivity, palpebral ptosis, catatonia and a stereotyped behaviour was produced by the extract when administered orally (1, 10 and 100 mg/kg). A significant decrease in respiration rate was observed (1, 10 and 100 mg/kg, p.o.) using a continuous flow respiration system. Lastly, doses of 1, 10 and 100 mg/kg, p.o. of the extract produced a significant increase in the hypnotic effect induced by pentobarbital in a dose dependent manner. The latest effects could probably explain its rational use in traditional medicine to alleviate stress or as a sedative agent. PMID- 10473173 TI - Evaluation of hepatoprotective activity of Cassia fistula leaf extract. AB - Hepatoprotective activity of the n-heptane extract of Cassia fistula leaves was investigated in rats by inducing hepatotoxicity with carbon tetrachloride:liquid paraffin (1:1). The extract has been shown to possess significant protective effect by lowering the serum levels of transminases (SGOT and SGPT), bilirubin and alkaline phosphatase (ALP). The extract of C. fistula at a dose of 400 mg/kg showed significant hepatoprotective activity which was comparable to that of a standard hepatoprotective agent. PMID- 10473175 TI - Screening of Turkish anti-ulcerogenic folk remedies for anti-Helicobacter pylori activity. AB - The anti-Helicobacter pylori effect of the extracts and fractions obtained from seven Turkish plants, which are used in folk medicine for the treatment of gastric ailments including peptic ulcers, were studied against one standard strain and eight clinical isolates of H. pylori by using the agar dilution method. Flowers of Cistus laurifolius and Spartium junceum, cones of Cedrus libani, herbs and flowers of Centaurea solstitialis ssp. solstitialis, fruits of Momordica charantia, herbaceous parts of Sambucus ebulus, and flowering herbs of Hypericum perforatum were evaluated in this study. Results showed that all except one extract from six of these plants showed activity against the microorganism with MICs between 1.95 and 250 microg/ml, with S. junceum being the only inactive species. Amongst the active plants the inhibitory properties of C. laurifolius were found prominent. PMID- 10473174 TI - Inhibition of immunoglobulin E production by Poncirus trifoliata fruit extract. AB - Immunoglobulin E (IgE) is the principal immunoglobulin involved in immediate hypersensitivities and chronic allergic diseases. The effect of an aqueous extract of Poncirus trifoliata (L) Raf. (Rutaceae) fruits (PTFE) on in vivo and in vitro IgE production was investigated. PTFE dose-dependently inhibited the active systemic anaphylaxis and serum IgE production induced by immunization with ovalbumin, Bordetella pertussis toxin and aluminum hydroxide gel. PTFE strongly inhibited interleukin 4 (IL-4)-dependent IgE production by lipopolysaccharide stimulated murine whole spleen cells. In the case of U266 human IgE-bearing B cells, PTFE also showed an inhibitory effect on the IgE production. These results suggest that PTFE has an anti-allergic activity by inhibition of IgE production from B cells. PMID- 10473176 TI - Inhibition of lipopolysaccharide plus substance P-induced tumor necrosis factor alpha production from astrocytes by Chongmyung-Tang. AB - Astrocytes have the capacity to secrete or respond to a variety of cytokines. In this study, we have examined whether an aqueous extract of Chongmyung-Tang (CmT) inhibits production of tumor necrosis factor-alpha (TNF-alpha) from primary cultures of mouse astrocytes. CmT (1 and 10 microg/ml) significantly inhibited the TNF-alpha production by astrocytes stimulated with lipopolysaccharide (LPS) and substance P (SP). Interleukin-1 (IL-1) has been shown to elevate TNF-alpha production from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. We therefore examined whether IL-1 mediated inhibition of TNF alpha production from primary astrocytes by CmT. Treatment of CmT (1 and 10 microg/ml) to astrocytes stimulated with both LPS and SP decreased IL-1 production significantly. In addition, reverse-transcribed polymerase chain reaction analysis demonstrated that significantly reduced level of the TNF-alpha mRNA was expressed in astrocytes treated with CmT. Our results suggest that CmT inhibits TNF-alpha production by reducing TNF-alpha mRNA expression. PMID- 10473177 TI - Apoptosis-associated generation of reactive oxygen intermediates and release of pro-inflammatory cytokines in human lymphocytes and granulocytes by extracts from the seeds of Acalypha wilkesiana. AB - Seeds from Acalypha wilkesiana (Euphorbiaceae) are an essential component of a complex plant mixture used empirically by traditional healers in Southwest Nigeria to treat breast tumours and inflammation. To investigate their biological properties, we incubated human lymphocytes and granulocytes with aqueous and ethanolic extracts of A. wilkesiana seeds (AWS). In lymphocytes, we observed an induction of apoptosis and generation of reactive oxygen intermediates (ROI), as measured by the oxidation of hydroethidine, within 2 h, while in granulocytes, an aqueous seed extract induced the oxidative burst and enhanced phagocytosis of Escherichia coli within 10-20 min. In the supernatants of 72-h cultured lymphocytes, AWS induced the release of the pro-inflammatory cytokines tumour necrosis factor-alpha and interleukin-6, and also T-cell-associated cytokines interleukin-5 and interferon-gamma. These preliminary results encourage further investigations of this drug with both cytotoxic and immunomodulating properties. PMID- 10473178 TI - Production of nitric oxide in mouse peritoneal macrophages after priming with interferon-gamma by the stem of Sinomenium acutum. AB - The present study demonstrates that the aqueous extract of Sinomenium acutum stem (SSAE) produces nitric oxide (NO) upon treatment with recombinant interferon gamma (rIFN-gamma) in mouse peritoneal macrophages. Apparently SSAE has no effect on NO production by itself. This production is dependent on L-arginine and can be inhibited by the L-arginine analogue N(G)-monomethyl-L-arginine. The increased production of NO from rIFN-gamma plus SSAE-stimulated cells was decreased by the treatment of protein kinase C inhibitor. Tumor necrosis factor-alpha (TNF-alpha) has been shown to stimulate the oxidative metabolism of L-arginine to produce NO. Mouse peritoneal macrophages secrete high levels of TNF-alpha after incubation with rIFN-gamma plus SSAE. In addition, SSAE-induced NO production is progressively inhibited by anti-murine TNF-alpha neutralizing antibody. These results show that the capacity of SSAE to increase NO production from rIFN-gamma primed mouse peritoneal macrophages is the result of SSAE-induced TNF-alpha secretion. PMID- 10473179 TI - Effect of Cryptotympana atrata extract on compound 48/80-induced anaphylactic reactions. AB - We studied the effect of the aqueous extract of Cryptotympana atrata Fabricius on anaphylactic reactions. Cryptotympana atrata completely inhibited compound 48/80 induced systemic anaphylactic shock at doses of 0.5 and 1.0 g/kg. When Cryptotympana atrata was given as pretreatment at concentrations ranging from 0.025 to 1.0 g/kg, the plasma histamine levels induced by compound 48/80 were reduced in a dose-dependent manner. Cryptotympana atrata also inhibited histamine release from rat peritoneal mast cells (RPMC) by compound 48/80. In addition, when Cryptotympana atrata was added, the level of cAMP in RPMC transiently and significantly increased about 14-fold compared with that of control cells. Our studies provide evidence that Cryptotympana atrata may be beneficial in the treatment of nonspecific anaphylactic reactions. PMID- 10473180 TI - In vitro evaluation of the amebicidal activity of Buddleia cordata (Loganiaceae, H.B.K.) on several strains of Acanthamoeba. AB - Infectious diseases produced by free-living amoebae from the genus Acanthamoeba have been recently recognized. The need for antiamebic compounds is urgent as the occurrence of these diseases is being registered more frequently since the late sixties. We screened the aqueous and methanolic extract of a plant used by folk medicine (Buddleia cordata) against eye and skin inflammation for antiamebic activity. We tested the extracts on 29 strains of free-living amoebae, with the result that they were amebostatic for 14 and 15 strains, respectively. We obtained linarin and vanillic acid from the extracts, but only linarin was amebostatic to all the strains and vanillic acid had no activity. However, acetyl vanillic acid had similar effects on amoebae to linarin. Threshold values of these two active compounds ranged from 31.25 microg/ml to 4 mg/ml and from 31.25 microg/ml to 8 mg/ml for linarin and acetyl vanillic acid, respectively. These differences in threshold values were observed even on several strains belonging to the same species (as in the case of A. castellanii and A. polyphaga) indicating the need of susceptibility testing for each clinical isolate of free living amoebae. PMID- 10473181 TI - Antifungal activities of seven West African Combretaceae used in traditional medicine. AB - Seven species of Combretaceae were investigated for their antifungal activity against the pathogenic fungi Candida albicans, Epidermophyton floccosum, Microsporum gypseum, Trichophyton mentagrophytes and Trichophyton rubrum. Pteleopsis suberosa and Terminalia avicennioides appear to be the most active plants. Phytochemical screening shows that these plants are particularly rich in tannins and saponins, which might be responsible for their antifungal activity. PMID- 10473182 TI - Antimicrobial activity of xanthones from Calophyllum species, against methicillin resistant Staphylococcus aureus (MRSA). AB - During the past 5 years, a considerable number of known and new xanthones from the Calophyllum species of Sri Lanka have been isolated and characterized. We have investigated the antimicrobial activity of Calophyllum xanthones, with a special reference to methicillin-resistant Staphylococcus aureus (MRSA). These activity studies were carried out using the agar plate method. Calozeloxanthone, a xanthone which has been isolated from C. moonii and C. lankensis, showed the highest activity against methicillin-resistant S. aureus (MRSA) strains at a concentration of 8.3 microg/ml. Hence, calozeyloxanthone appears to hold promise as an antimicrobial agent in the treatment of infections with S. aureus, including methicillin-sensitive S. aureus (MRSA), and should be investigated further. PMID- 10473183 TI - Occurrence of usnic acid in Usnea laevis Nylander (lichenized ascomycetes) from the Venezuelan Andes. AB - The presence of usnic acid in the lichen Usnea laevis Nyl. from the Venezuelan Andes was detected through chromotographic (TLC) and spectroscopic (IR, MS, 1H NMR) methods. This compound was present in a concentration of 2.7% in the thallus. Usnic acid has a reported antibiotic activity and the lichen is utilized for medicinal purposes by Andean farmers. PMID- 10473184 TI - In vitro inhibition of drug-resistant and drug-sensitive strains of Mycobacterium tuberculosis by ethnobotanically selected South African plants. AB - Twenty South African medicinal plants used to treat pulmonary diseases were screened for activity against drug-resistant and drug-sensitive strains of Mycobacterium tuberculosis. A preliminary screening of acetone and water plant extracts against a drug-sensitive strain of Mycobacterium tuberculosis, H37Rv, was done by the agar plate method. Fourteen of the 20 acetone extracts showed inhibitory activity at a concentration of 0.5 mg/ml against this strain. Acetone as well as water extracts of Cryptocarya latifolia, Euclea natalensis, Helichrysum melanacme, Nidorella anomala and Thymus vulgaris inhibited the growth of M. tuberculosis. Given the activity of 14 acetone extracts at 0.5 mg/ml against the drug-sensitive strain by the agar plate method, a further study was done employing a rapid radiometric method to confirm the inhibitory activity. These active acetone extracts were screened against the H37Rv strain as well as a strain resistant to the drugs isoniazid and rifampin. The minimal inhibitory concentration of Croton pseudopulchellus, Ekebergia capensis, Euclea natalensis, Nidorella anomala and Polygala myrtifolia was 0.1 mg/ml against the H37Rv strain by the radiometric method. Extracts of Chenopodium ambrosioides, Ekebergia capensis, Euclea natalensis, Helichrysum melanacme, Nidorella anomala and Polygala myrtifolia were active against the resistant strain at 0.1 mg/ml. Eight plants showed activity against both strains at a concentration of 1.0 mg/ml. PMID- 10473185 TI - Hepatoprotective activity of leaves of Cassia occidentalis against paracetamol and ethyl alcohol intoxication in rats. AB - Cassia occidentalis L. (Caesalpiniaceae), commonly known as 'Kasondi', is used in Unani medicine for liver ailments and is an important ingredient of several polyherbal formulations marketed for liver diseases. The hepatoprotective effect of aqueous-ethanolic extract (50%, v/v) of leaves of kasondi was studied on rat liver damage induced by paracetamol and ethyl alcohol by monitoring serum transaminase (aspartate amino transferase and serum alanine amino transferase), alkaline posphatase, serum cholesterol, serum total lipids and histopathological alterations. The extract of leaves of the plant produced significant hepatoprotection. PMID- 10473186 TI - Assays for noninvasive imaging of reporter gene expression. AB - Repeated, noninvasive imaging of reporter gene expression is emerging as a valuable tool for monitoring the expression of genes in animals and humans. Monitoring of organ/cell transplantation in living animals and humans, and the assessment of environmental, behavioral, and pharmacologic modulation of gene expression in transgenic animals should soon be possible. The earliest clinical application is likely to be monitoring human gene therapy in tumors transduced with the herpes simplex virus type 1 thymidine kinase (HSV1-tk) suicide gene. Several candidate assays for imaging reporter gene expression have been studied, utilizing cytosine deaminase (CD), HSV1-tk, and dopamine 2 receptor (D2R) as reporter genes. For the HSV1-tk reporter gene, both uracil nucleoside derivatives (e.g., 5-iodo-2'-fluoro-2'-deoxy-1-beta-D-arabinofuranosyl-5-iodouracil [FIAU] labeled with 124I, 131I) and acycloguanosine derivatives [e.g., 8-[18F]fluoro-9 [[2-hydroxy-1-(hydroxymethyl)ethoxy]methyl]guanine (8-[18F]-fluoroganciclovir) ([18F]FGCV), 9-[(3-[18F]fluoro-1-hydroxy-2-propoxy)methyl]guanine ([18F]FHPG)] have been investigated as reporter probes. For the D2R reporter gene, a derivative of spiperone [3-(2'-[18F]-Fluoroethyl)spiperone ([18F]FESP)] has been used with positron emission tomography (PET) imaging. In this review, the principles and specific assays for imaging reporter gene expression are presented and discussed. Specific examples utilizing adenoviral-mediated delivery of a reporter gene as well as tumors expressing reporter genes are discussed. PMID- 10473187 TI - PET examination of three potent cocaine derivatives as specific radioligands for the serotonin transporter. AB - Several positron emission tomography (PET) radioligands based on the aryl tropane structure have been used for studies on monoamine reuptake sites. RTI-364, RTI 330, and RTI-357 (3-beta-(4'-n-propyl-,4'-iso-propyl-, and 4'-iso-propenyl phenyl)nortropane-2-beta-carboxylic acid methyl ester) are three recently synthesized cocaine analogues with higher affinity for the serotonin (5-HTT) than the dopamine transporter (DAT). Unlabelled RTI-364 and RTI-330 were prepared in a two-step synthesis. The key step was the addition of the appropriate propyl Grignard reagent to anhydroecgonine methyl ester. RTI-357 was prepared in a three step synthesis with a palladium-catalyzed coupling reaction of beta-CIT and isopropenylzinc bromide as key step. Hydrolysis of the ester functions gave the carboxylic acid analogues of RTI-364, RTI-330, and RTI-357, which were labelled with 11C using [11C]methyl iodide in dimethyl formamide (DMF) and tetrabutylammonium hydroxide (TBAH) as base. All three compounds entered the monkey brain in a high degree (approximately 5-10%). There was a low uptake of [11C]RTI-364 in serotonin-rich brain areas, whereas [11C]RTI-330 and [11C]RTI-357 showed a marked uptake of radioactivity in the thalamus and the brainstem, regions known to contain serotonin transporters. Transient equilibrium was reached at 15 and 40 min for [11C]RTI-330 and [11C]RTI-357, respectively. After pretreatment with citalopram, the ratio of radioactivity in the thalamus and the brainstem to the cerebellum were markedly reduced for [11C]RTI-357 but not for [11C]RTI-330. The results indicate that [11C]RTI-357 is a potential PET radioligand for quantitation of the serotonin reuptake site. PMID- 10473188 TI - Characterization of bromine-76-labelled 5-bromo-6-nitroquipazine for PET studies of the serotonin transporter. AB - The development of suitable radioligands for brain imaging of the serotonin transporter is of great importance for the study of depression and other affective disorders. The potent and selective serotonin transporter ligand, 5 iodo-6-nitro-2-piperazinylquinoline, has been labelled with iodine-123 and used as a radioligand for single photon emission computerized tomography. To evaluate the potential of the bromine-76-labelled analogue, 5-bromo-6-nitroquipazine, as a radioligand for positron emission tomography (PET), its brain distribution and binding characteristics were examined in rats. In vivo brain distribution and ex vivo autoradiography demonstrated that [76Br]5-bromo-6-nitroquipazine enters the brain rapidly. The regional brain distribution of [76Br]5-bromo-6-nitroquipazine was consistent with the known distribution of serotonin transporters in the midbrain, pons, thalamus, striatum, and neocortex. Specific binding was inhibited by the selective serotonin reuptake inhibitor citalopram. The peripheral metabolism in plasma was rapid, but more than 90% of the radioactivity in brain represented unchanged radioligand 2 h postinjection (p.i.). A preliminary PET study was also performed in a baboon. Following the intravenous injection of [76Br]5-bromo-6-nitroquipazine in a baboon, there was a conspicuous accumulation of radioactivity in thalamus, striatum, and pons. The radioactivity in these brain regions was 1.5 times higher than in the cerebellum at 3 h and 2.5-4 times higher at 24 h. A rapid metabolism of the radioligand in plasma was observed (38% unchanged after 5 min). The results indicate that [76Br]5-bromo-6-nitroquipazine has potential for PET imaging of the serotonin transporter. PMID- 10473189 TI - Carbon-11 epidepride: a suitable radioligand for PET investigation of striatal and extrastriatal dopamine D2 receptors. AB - Epidepride [(S)-(-)-N-([1-ethyl-2-pyrrolidinyl]methyl)-5-iodo-2,3-dimethoxybenza mide] binds with a picomolar affinity (Ki = 24 pM) to the dopamine D2 receptor. Iodine-123-labeled epidepride has been used previously to study striatal and extrastriatal dopamine D2 receptors with single photon emission computed tomography (SPECT). Our aim was to label epidepride with carbon-11 for comparative quantitative studies between positron emission tomography (PET) and SPECT. Epidepride was synthesized from its bromo-analogue FLB 457 via the corresponding trimethyl-tin derivative. In an alternative synthetic pathway, the corresponding substituted benzoic acid was reacted with the optically pure aminomethylpyrrolidine-derivative. Demethylation of epidepride gave the desmethyl derivative, which was reacted with [11C]methyl triflate. Total radiochemical yield was 40-50% within a total synthesis time of 30 min. The specific radioactivity at the end of synthesis was 37-111 GBq/micromol (1,000-3,000 Ci/mmol). Human postmortem whole-hemisphere autoradiography demonstrated dense binding in the caudate putamen, and also in extrastriatal areas such as the thalamus and the neocortex. The binding was inhibited by unlabeled raclopride. PET studies in a cynomolgus monkey demonstrated high uptake in the striatum and in several extrastriatal regions. At 90 min after injection, uptake in the striatum, thalamus and neocortex was about 11, 4, and 2 times higher than in the cerebellum, respectively. Pretreatment experiment with unlabeled raclopride (1 mg/kg) inhibited 50-70% of [11C]epidepride binding. The fraction of unchanged [11C]epidepride in monkey plasma determined by a gradient high performance liquid chromatography (HPLC) method was about 30% of the total radioactivity at 30 min after injection of [11C]epidepride. The availability of [11C]epidepride allows the PET-verification of the data obtained from quantitation studies with SPECT. PMID- 10473190 TI - Preliminary assessment of extrastriatal dopamine D-2 receptor binding in the rodent and nonhuman primate brains using the high affinity radioligand, 18F fallypride. AB - We have identified the value of 18F-fallypride [(S)-N-[(1-allyl-2 pyrrolidinyl)methyl]-5-(3-[18F]fluoropropyl)-2, 3-dimethoxybenzamide], as a dopamine D-2 receptor radiotracer for the study of striatal and extrastriatal receptors. Fallypride exhibits high affinities for D-2 and D-3 subtypes and low affinity for D-4 (3H-spiperone IC50s: D-2 = 0.05 nM [rat striata], D-3 = 0.30 nM [SF9 cell lines, rat recombinant], and D-4 = 240 nM [CHO cell lines, human recombinant]). Biodistribution in the rat brain showed localization of 18F fallypride in striata and extrastriatal regions such as the frontal cortex, parietal cortex, amygdala, hippocampus, thalamus, and hypothalamus. In vitro autoradiographic studies in sagittal slices of the rat brain showed localization of 18F-fallypride in striatal and several extrastriatal regions, including the medulla. Positron emission tomography (PET) experiments with 18F-fallypride in male rhesus monkeys were carried out in a PET VI scanner. In several PET experiments, apart from the specific binding seen in the striatum, specific binding of 18F-fallypride was also identified in extracellular regions (in a lower brain slice, possibly the thalamus). Specific binding in the extrastriata was, however, significantly lower compared with that observed in the striata of the monkeys (extrastriata/cerebellum = 2, striata/cerebellum = 10). Postmortem analysis of the monkey brain revealed significant 18F-fallypride binding in the striata, whereas binding was also observed in extrastriatal regions such as the thalamus, cortical areas, and brain stem. PMID- 10473191 TI - Synthesis of 3beta-(4-[18F]fluoromethylphenyl)- and 3beta-(2-[18F] fluoromethylphenyl)tropane-2beta-carboxylic acid methyl esters: new ligands for mapping brain dopamine transporter with positron emission tomography. AB - The synthesis of two new dopamine transporter ligands, 3beta-(4 fluoromethylphenyl)tropane-2beta-carboxylic acid methyl ester and 3beta-(2 fluoromethylphenyl)tropane-2beta-carboxylic acid methyl ester, and their spectral characterization are described. The precursors for these ligands were prepared by TiCl4 catalyzed chloromethylation of 3beta-phenyltropane-2beta-carboxylic acid methyl ester followed by separation of the isomeric product mixture of 2- and 4 chloromethylphenyltropane derivatives. Reaction of the chloromethyl analogs with no-carrier-added [18F]fluoride ion followed by high performance liquid chromatography purification provided the corresponding [18F]fluoromethyltropanes, in good radiochemical yields, useful for imaging the brain dopamine transporter system in vivo with positron emission tomography. PMID- 10473192 TI - Dopamine D1 agonist R-[11C]SKF 82957: synthesis and in vivo characterization in rats. AB - The active enantiomer R-SKF 82957 was labeled with 11C by N-[11C]methylation of the full dopamine (D1) agonist R-SKF 81297, using [11C]methyl iodide in the presence of N-ethyldiisopropylamine, in high specific activity, radiochemical purity and yields. Compared with the D1 agonist R/S-[11C]SKF 82957, R-[11C]SKF 82957 showed higher binding in the D1 rich regions, such as striatum and olfactory tubercles (approximately 1.7 times), thereby improving the tissue contrast. R-[11C]SKF 82957 exhibited high in vivo binding selectivity for D1 receptors in rats, because only high doses of D1 competitors, but not D2 or serotonin (5-HT2) blockers, significantly reduced the radioactivity levels in all brain areas. No labeled metabolites were detected in rat brain. These results indicate that R-[11C]SKF 82957 will provide more sensitive measurements of D1 receptors in in vivo studies than the racemic mixture. PMID- 10473193 TI - Simplified methods for in vivo measurement of acetylcholinesterase activity in rodent brain. AB - Simplified methods for in vivo studies of acetylcholinesterase (AChE) activity in rodent brain were evaluated using N-[11C]methylpiperidinyl propionate ([11C]PMP) as an enzyme substrate. Regional mouse brain distributions were determined at 1 min (representing initial brain uptake) and 30 min (representing trapped product) after intravenous [11C]PMP administration. Single time point tissue concentrations (percent injected dose/gram at 30 min), tissue concentration ratios (striatum/cerebellum and striatum/cortex ratios at 30 min), and regional tissue retention fractions (defined as percent injected dose 30 min/percent injected dose 1 min) were evaluated as measures of AChE enzymatic activity in mouse brain. Studies were carried out in control animals and after dosing with phenserine, a selective centrally active AChE inhibitor; neostigmine, a peripheral cholinesterase inhibitor; and a combination of the two drugs. In control and phenserine-treated animals, absolute tissue concentrations and regional retention fractions provide good measures of dose-dependent inhibition of brain AChE; tissue concentration ratios, however, provide erroneous conclusions. Peripheral inhibition of cholinesterases, which changes the blood pharmacokinetics of the radiotracer, diminishes the sensitivity of all measures to detect changes in central inhibition of the enzyme. We conclude that certain simple measures of AChE hydrolysis rates for [11C]PMP are suitable for studies where alterations of the peripheral blood metabolism of the tracer are kept to a minimum. PMID- 10473194 TI - Synthesis and evaluation of copper radiopharmaceuticals with mixed bis(thiosemicarbazone) ligands. AB - Four "mixed" bis(thiosemicarbazone) derivatives of pyruvaldehyde were synthesized that incorporate two dissimilar thiosemicarbazone functions. The corresponding [67Cu]copper(II) complexes were prepared and evaluated as possible copper radiopharmaceuticals. The pyruvaldehyde-based mixed bis(thiosemicarbazone) ligands, CH3C[=NNHC(S)NHMe]CH[=NNHC(S)NHEt] (1), CH3C[=NNHC(S)NHMe] CH[=NNHC(S)NEt2] (2), CH3C[=NNHC(S)NHMe]CH[=NNHC(S)-cyclo-N(CH2)5] (3), and CH3C [=NNHC(S)NHMe]CH[=NNHC(S)-cyclo-N(CH2)6] (4), were obtained by reaction of the appropriate thiosemicarbazide derivative with pyruvaldehyde-2-N4 methylthiosemicarbazone (CH3C[=NNHC(S) NHMe]CHO). The 67Cu-labeled copper(II) complexes of ligands 1-4 were prepared and screened in a rat model to assess the potential of each chelate as a 62Cu-radiopharmaceutical for imaging with positron emission tomography. The 67Cu-complexes of ligands 1-4 exhibit significant uptake into the brain and heart 1 min following intravenous administration to rats. For the 67Cu-complexes of ligands 2, 3, and 4, the cerebral and myocardial uptake of 67Cu is two-to-threefold lower at 2 h than at 1 min postinjection, due to significant biological clearance of these 67Cu-chelates. However, the 67Cu complex of 1 affords cerebral and myocardial uptake and retention comparable to that of [67Cu]Cu-PTSM in this model. Although the kinetics of this new agent appear attractive, ultrafiltration studies using solutions of dog and human serum albumin reveal that the 67Cu-complex of ligand 1, like Cu-PTSM, interacts more strongly with human albumin than dog albumin. Thus, this new agent would appear to offer no advantage over Cu-PTSM as a 62Cu-labeled tracer for evaluation of regional tissue perfusion. PMID- 10473195 TI - Radiochemical studies of 99mTc complexes of modified cysteine ligands and bifunctional chelating agents. AB - The synthesis of four novel ligands using the amino-acid cysteine and its ethyl carboxylate derivative is described. The synthetic method involves a two-step procedure, wherein the intermediate Schiff base formed by the condensation of the amino group of the cysteine substrate and salicylaldehyde is reduced to give the target ligands. The intermediates and the final products were characterized by high resolution nuclear magnetic resonance spectroscopy. Complexation studies of the ligands with 99mTc were optimized using stannous tartrate as the reducing agent under varying reaction conditions. The complexes were characterized using standard quality control techniques such as thin layer chromatography, paper electrophoresis, and paper chromatography. Lipophilicities of the complexes were estimated by solvent extraction into chloroform. Substantial changes in net charge and lipophilicity of the 99mTc complexes were observed on substituting the carboxylic acid functionality in ligands I and II with the ethyl carboxylate groups (ligands II and IV). All the ligands formed 99mTc complexes in high yield. Whereas the complexes with ligands I and II were observed to be hydrophilic in nature and not extractable into CHCl3, ligands III and IV resulted in neutral and lipophilic 99mTc complexes. The 99mTc complex with ligand II was not stable and on storage formed a hydrophilic and nonextractable species. The biodistribution of the complexes of ligands I and II showed that they cleared predominantly through the kidneys, whereas the complexes with ligands III and IV were excreted primarily through the hepatobiliary system. No significant brain uptake was observed with the 99mTc complexes with ligands III and IV despite their favorable properties of neutrality, lipophilicity, and conversion into a hydrophilic species. These ligands offer potential for use as bifunctional chelating agents. PMID- 10473196 TI - 99mTc(L-L)3+ complexes containing ether analogs of DMPE. AB - Novel 99mTc(L-L)3+ complexes have been investigated for potential use in myocardial perfusion imaging. Bidentate chelators have been prepared that are based on substituent analogs of 1,2-bis(dimethylphosphino)ethane, onto which alkyl ether groups have been incorporated. The new ligands are: (1) MMPE, 1,2 bis(methyl methoxyethyl phosphino)ethane, (2) MIBPE, 1,2-bis(methyl methoxyisobutyl phosphino)ethane, (3) FURPE, 1,2-bis(methyl tetrahydrofuran phosphino)ethane, and (4) PYRPE, 1,2-bis(methyl tetrahydropyran phosphino)ethane. These ligands have been reacted with 99mTc and the resulting complexes evaluated for myocardial imaging properties. 99mTc(MMPE)3+ exhibited the most favorable myocardial imaging characteristics in animal models. Results indicate that pendent ether moieties can improve the myocardial imaging properties of cationic technetium complexes. PMID- 10473197 TI - [188Re]Rhenium sulfide suspension: a potential radiopharmaceutical for tumor treatment following intra-tumor injection. AB - Intralesional therapy has been shown to be an effective treatment for tumors. In this study, the suitability of [188Re]rhenium sulfide suspension for tumor treatment following intra-tumor injection was evaluated. The [188Re]rhenium sulfide suspension was radiolabeled with 188Re with a radiochemical yield of more than 96%. In vitro stability studies revealed that more than 99% of the 188Re remained in sulfide form over a 3-day period. After ultrasonication for 5 or 10 min, the main particle size was 1-5 microm. Two [188Re]rhenium sulfide suspensions ultrasonicated for 5 and 10 min, respectively, were injected into separate group of tumor-bearing mice that were killed after specified times to compare the retention of 188Re in tumors and the leakage to different organs by periods and organs removed to gamma counting. The mean retention percentages of 188Re in tumors injected with suspension ultrasonicated for 5 (or 10) min were as follows: 1 h, 90.5 +/- 7.7% (83.1 +/- 13.7%); 24 h, 92.2 +/- 8.6% (83.9 +/- 9.8%); 48 h, 88.3 +/- 10.9% (80.2 +/- 3.8%); and 72 h, 91.5 +/- 7.6% (78.8 +/- 3.0%). Tumor-inhibiting ratio was 96.5%. These results demonstrated that [188Re]rhenium sulfide suspension is an effective radiopharmaceutical for tumor treatment by intralesional therapy. PMID- 10473198 TI - Comparison of 225actinium chelates: tissue distribution and radiotoxicity. AB - The biodistribution and tissue toxicity of intravenously administered 225 actinium (225Ac) complexed with acetate, ethylene diamine tetraacetic acid (EDTA), 1, 4, 7, 10, 13-pentaazacyclopentadecane-N, N', N", N"', N""-pentaacetic acid (PEPA), or the "a" isomer of cyclohexyl diethylenetriamine pentaacetic acid (CHX-DTPA), were examined. The percent of injected dose per organ and per gram of tissue for each chelate complex was determined. 225Ac-CHX-DTPA was evaluated further for radiotoxic effects. Mice receiving > or =185 kBq 225Ac-CHX-DTPA suffered 100% morbidity by 5 days and 100% mortality by 8 days postinjection, and all animals evaluated had significant organ damage. The in vivo instability of the 225Ac-CHX-DTPA complex likely allowed accumulation of free 225Ac in organs, which resulted in tissue pathology. PMID- 10473199 TI - Radiolabelling of 4-iodo-N-(2-morpholinoethyl)benzamide with NA123I and NA125I. PMID- 10473200 TI - Dynamics of polymerization in polyacrylamide gel (PAG) dosimeters: (I) ageing and long-term stability. AB - Few quantitative data are available on the kinetics of polymerization reactions in polymer gel (PAG) dosimeters and their long-term stability. Post-irradiation polymerization reactions have been found to continue for several weeks, posing questions regarding dosimeter stability and its achievement. In this paper we report an investigation of polymerization kinetics in PAG dosimeters and the effect of diffusing oxygen into the dosimeter, post irradiation, as a potential method of inhibiting further polymerization and stabilizing the dose distribution. Results show continuous post-irradiation changes in transverse relaxation rate R2 with time over the five week study period and that a steady state may not be reached for a period of months. An assessment is made of the appropriate time to image the dosimeter which shows that after three to four days the polymerization change is slow compared with imaging time. The implications of the time delay between irradiation and imaging are discussed in terms of the resultant sensitivity of the dosimeter and accuracy of the dose measured. In pairs of dosimeters, one sealed the other open to air, oxygen diffusing into the dosimeter arrests polymerization. However, the diffusion rate is too slow to make this method practicable. The slow diffusion means that while in regions near the air/gel interface polymerization is quickly arrested, in deeper regions it may continue for many hours, causing artefacts in the dose distribution. In the companion paper to this from a collaborating team, a study focusing on modelling oxygen diffusion in dosimeter gel will be presented. PMID- 10473201 TI - Dynamics of polymerization in polyacrylamide gel (PAG) dosimeters: (II) modeling oxygen diffusion. AB - Oxygen contamination of a polyacrylamide gel (PAG) dosimeter can have a significant detrimental effect upon the performance characteristics of the gel as a dosimeter. Oxygen contamination can occur during preparation, but can also arise from the diffusion of atmospheric oxygen through vessel walls. Even 'trace' amounts of oxygen present in any gaseous space (usually nitrogen filled) between the surface of the PAG and the vessel seal can have a detectable effect. This paper describes a method used to quantify the detrimental effects of a range of oxygen concentrations upon the performance of the PAG. The results of diffusion studies for air and pure oxygen diffusing into the PAG over time are presented and coupled with a novel method of calculating oxygen concentrations in the PAG. Results obtained suggest that the diffusion is Fickian. The coefficient of diffusion D has been calculated to be (8+/-2) x 10(-6) cm2 s(-1) for oxygen diffusing into PAG stored at 4 degrees C, under the assumption that the diffusion mechanism is independent of the concentration of the diffusing species. In addition, a quantitative relation has been established between the transverse relaxation rate R2 of the PAG and the concentration of oxygen present. The implications of these findings for polymer gel dosimetry are discussed. PMID- 10473202 TI - Monte Carlo-based inverse treatment planning. AB - A Monte Carlo based inverse treatment planning system (MCI) has been developed which combines arguably the most accurate dose calculation method (Monte Carlo particle transport) with a 'guaranteed' optimization method (simulated annealing). A distribution of photons is specified in the tumour volume; they are transported using an adjoint calculation method to outside the patient surface to build up an intensity distribution. This intensity distribution is used as the initial input into an optimization algorithm. The dose distribution from each beam element from a number of fields is pre-calculated using Monte Carlo transport. Simulated annealing optimization is then used to find the weighting of each beam element, to yield the optimal dose distribution for the given criteria and constraints. MCI plans have been generated in various theoretical phantoms and patient geometries. These plans show conformation of the dose to the target volume and avoidance of critical structures. To verify the code, an experiment was performed on an anthropomorphic phantom. PMID- 10473203 TI - Monte Carlo calculations of the ionization chamber wall correction factors for 192Ir and 60Co gamma rays and 250 kV x-rays for use in calibration of 192Ir HDR brachytherapy sources. AB - As in the method for the calibration of 192Ir high-dose-rate (HDR) brachytherapy sources, the ionization chamber wall correction factor A(w), is needed for 192Ir and 60Co gamma rays and 250 kV x-rays. This factor takes into account the variation in chamber response due to the attenuation of the photon beam in the chamber wall and build-up cap and the contribution of scattered photons. Monte Carlo calculations were performed using the EGS4 code system with the PRESTA algorithm, to calculate the A(w) factor for 51 commercial ionization chambers and build-up caps exposed to the typical energy spectrum of 192Ir and 60Co gamma rays and 250 kV x-rays. The calculated A(w) correction factors for 192Ir and 60Co sources and 250 kV x-rays agree very well to within 0.1% with published experimental data (the statistical uncertainty is less than 0.1% of the calculated correction factor value). For the 192Ir sources, A(w) varies from 0.973 to 0.993 and for the 250 kV x-rays the minimum value of A(w) for all chambers studied is 0.983. The calculated A(w) correction factors can be used to calculate the air kerma calibration factor of HDR brachytherapy sources, when interpolative methods are considered, contributing to the reduction in the overall uncertainties in the calibration procedure. PMID- 10473204 TI - An improved technique for comparing Gamma Knife dose-volume distributions in stereotactic radiosurgery. AB - A function derived from the geometry of brachytherapy dose distributions is applied to stereotactic radiosurgery and an algorithm for the production of a novel dose-volume histogram, the Anderson inverse-square shifted dose-volume histogram (DVH), is proposed. The expected form of the function to be plotted is checked by calculating its value for single focus exposures, and its application to clinical examples of Gamma Knife treatments described. The technique is shown to provide a valuable tool for assessing the adequacy of radiosurgical plans and comparing and reporting dose distributions. PMID- 10473205 TI - Validation of K-edge 125I brachytherapy enhancement with silver compounds. AB - Brachytherapy with radioactive seeds implanted within the tumour volume has demonstrated good success rates in treating certain cancers. In an effort to improve the curative rates in cancer patients, ongoing research is being conducted to enhance the amount of radiation dose that is absorbed within the tumour volume while minimizing the dose absorbed by the surrounding normal tissue. One method for enhancing tumour dose absorption with 125I brachytherapy seeds is to increase the number of photoelectric atomic interactions within the tumour volume by introducing small quantities of a silver compound, taking advantage of the K-edge effect. Because low-energy electrons and Auger electrons are the primary sources of brachytherapy dose enhancement, acquiring accurate experimental measurements of absorbed dose increases is a major challenge. To circumvent this problem, an x ray fluorescence excitation spectroscopy dosimetry technique supplemented with clinically accepted dosimetry calculations was developed to estimate relative absorbed dose increases in a water phantom containing up to 7.5 mM of silver. Excellent agreement was observed between theoretically derived Monte Carlo dosimetric predictions and experimental measurements. These results successfully demonstrated that K-edge enhanced 125I brachytherapy is indeed possible with future development of a non-toxic silver chelate. PMID- 10473206 TI - Sensitivity of coefficients for converting entrance surface dose and kerma-area product to effective dose and energy imparted to the patient. AB - We investigate the sensitivity of the conversions from entrance surface dose (ESD) or kerma-area product (KAP) to effective dose (E) or to energy imparted to the patient (epsilon) to the likely variations in tube potential, field size, patient size and sex which occur in clinical work. As part of a factorial design study for chest and lumbar spine examinations, the tube potentials were varied to be +/-10% of the typical values for the examinations while field sizes and the positions of the field centres were varied to be representative of values drawn from measurements on patient images. Variation over sex and patient size was based on anthropomorphic phantoms representing males and females of ages 15 years (small adult) and 21 years (reference adult). All the conversion coefficients were estimated using a mathematical phantom programmed with the Monte Carlo code EGS4 for all factor combinations and analysed statistically to derive factor effects. In general, the factors studied behaved independently in the sense that interaction of the physical factors generally gave no more than a 5% variation in a conversion coefficient. Taken together, variation of patient size, sex, field size and field position can lead to significant variation of E/KAP by up to a factor of 2, of E/ESD by up to a factor of 3, of epsilon/KAP by a factor of 1.3 and of epsilon/ESD by up to a factor of 2. While KAP is preferred to determine epsilon, the results show no strong preference of KAP over ESD in determining E. The mean absorbed dose D in the patient obtained by dividing epsilon (determined using KAP) by the patient's mass was found to be the most robust measure of E. PMID- 10473207 TI - Dose distributions using kilovoltage x-rays and dose enhancement from iodine contrast agents. AB - In x-ray phototherapy of brain tumours, the tumour is loaded with iodine and exposed to kilovoltage x-rays. Due to the high photoelectric cross sections of iodine, substantial photoelectric interactions occur. The flux of photoelectrons, characteristic x-rays and Auger electrons produce a localized dose enhancement. A modified computed tomography scanner, CTRx, can be used both for tumour localization and delivery of the dose enhancement therapy. Monte Carlo methods were employed to simulate the treatment of iodinated brain tumours with a CTRx. The calculated results reveal the effect of tumour iodine concentration on dose distribution, the degree of skull bone sparing with the application of multiple arcs, and the homogeneity of tumour dose distribution versus iodine concentration. A comparison with 10 MV stereotactic radiosurgery treatment shows the potential of CTRx treatment relative to conventional treatment modalities. PMID- 10473208 TI - Source localization in an inhomogeneous physical thorax phantom. AB - The influence of lung inhomogeneities on focal source localizations in electrocardiography (ECG) and magnetocardiography (MCG) is investigated. A realistically shaped physical thorax phantom with cylindrical lung inhomogeneities is used for electric and magnetic measurements. The lungs are modelled with a special ionic exchange membrane which allows different conductivity compartments without influencing the free ionic current flow. The dipolar current sources are composed of platinum wire and located at different depths and directions between the lung inhomogeneities. We localized the current dipoles with different boundary element method (BEM) models, based on electrical data and simultaneous electrical and magnetic data. Our results indicate the possibility of superadditive information gain by combining electrical and magnetic data for source reconstructions. We found a significant influence of the inhomogeneities on both the calculated source location and the calculated source strength. Mislocalizations of up to 16 mm and wrong dipole strengths of up to 52% were obtained when the lung inhomogeneities were not taken into account for source localization. Dipoles parallel to the lungs showed a larger localization error in depth than dipoles perpendicular to the lungs. We conclude that the incorporation of lung inhomogeneities will improve source localization accuracy in ECG and MCG. PMID- 10473209 TI - Ferrous sulphate gel dosimetry and MRI for proton beam dose measurements. AB - Ferrous sulphate gel dosimetry has the potential for measurement of absorbed dose distributions in proton therapy. The chemical properties of the gel are altered according to the radiation dose and these changes can be evaluated in three dimensions using MRI. The purpose of this work was to investigate the properties of a ferrous gel used with clinical proton beams. The gel was irradiated with both monoenergetic and range-modulated proton beams. It was then evaluated using MRI. The depth dose by means of the 1/T1 distribution was studied and compared with data from a plane-parallel plate ionization chamber. 1/T1 was shown to be proportional to the dose at a mean proton energy of approximately 90 MeV. The dose response was no different from that obtained using photon beams. However, on normalization at the entrance, the relative 1/T1 at the Bragg peak was 15-20% lower than the corresponding ionization chamber data for the monoenergetic proton beam. Better agreement was found for the modulated beam, but with significant differences close to the distal edge of the 1/T1 distribution. The change in sensitivity with depth was explained by means of a linear energy transfer dependence. This property was further studied using Monte Carlo methods. PMID- 10473210 TI - Analytical propagation of errors in dynamic SPECT: estimators, degrading factors, bias and noise. AB - Dynamic SPECT is a relatively new technique that may potentially benefit many imaging applications. Though similar to dynamic PET, the accuracy and precision of dynamic SPECT parameter estimates are degraded by factors that differ from those encountered in PET. In this work we formulate a methodology for analytically studying the propagation of errors from dynamic projection data to kinetic parameter estimates. This methodology is used to study the relationships between reconstruction estimators, image degrading factors, bias and statistical noise for the application of dynamic cardiac imaging with 99mTc-teboroxime. Dynamic data were simulated for a torso phantom, and the effects of attenuation, detector response and scatter were successively included to produce several data sets. The data were reconstructed to obtain both weighted and unweighted least squares solutions, and the kinetic rate parameters for a two-compartment model were estimated. The expected values and standard deviations describing the statistical distribution of parameters that would be estimated from noisy data were calculated analytically. The results of this analysis present several interesting implications for dynamic SPECT. Statistically weighted estimators performed only marginally better than unweighted ones, implying that more computationally efficient unweighted estimators may be appropriate. This also suggests that it may be beneficial to focus future research efforts upon regularization methods with beneficial bias-variance trade-offs. Other aspects of the study describe the fundamental limits of the bias variance trade-off regarding physical degrading factors and their compensation. The results characterize the effects of attenuation, detector response and scatter, and they are intended to guide future research into dynamic SPECT reconstruction and compensation methods. PMID- 10473211 TI - A study of artefacts in simultaneous PET and MR imaging using a prototype MR compatible PET scanner. AB - We have assessed the possibility of artefacts that can arise in attempting to perform simultaneous positron emission tomography (PET) and magnetic resonance imaging (MRI) using a small prototype MR compatible PET scanner (McPET). In these experiments, we examine MR images for any major artefacts or loss in image quality due to inhomogeneities in the magnetic field, radiofrequency interference or susceptibility effects caused by operation of the PET system inside the MR scanner. In addition, possible artefacts in the PET images caused by the static and time-varying magnetic fields or radiofrequency interference from the MR system were investigated. Biological tissue and a T2-weighted spin echo sequence were used to examine susceptibility artefacts due to components of the McPET scanner (scintillator, optical fibres) situated in the MR field of view. A range of commonly used MR pulse sequences was studied while acquiring PET data to look for possible artefacts in either the PET or MR images. Other than a small loss in signal-to-noise using gradient echo sequences, there was no significant interaction between the two imaging systems. Simultaneous PET and MR imaging of simple phantoms was also carried out in different MR systems with field strengths ranging from 0.2 to 4.7 T. The results of these studies demonstrate that it is possible to acquire PET and MR images simultaneously, without any significant artefacts or loss in image quality, using our prototype MR compatible PET scanner. PMID- 10473212 TI - Image/patient registration from (partial) projection data by the Fourier phase matching method. AB - A technique for 2D or 3D image/patient registration, PFPM (projection based Fourier phase matching method), is proposed. This technique provides image/patient registration directly from sequential tomographic projection data. The method can also deal with image files by generating 2D Radon transforms slice by slice. The registration in projection space is done by calculating a Fourier invariant (FI) descriptor for each one-dimensional projection datum, and then registering the FI descriptor by the Fourier phase matching (FPM) method. The algorithm has been tested on both synthetic and experimental data. When dealing with translated, rotated and uniformly scaled 2D image registration, the performance of the PFPM method is comparable to that of the IFPM (image based Fourier phase matching) method in robustness, efficiency, insensitivity to the offset between images, and registration time. The advantages of the former are that subpixel resolution is feasible, and it is more insensitive to image noise due to the averaging effect of the projection acquisition. Furthermore, the PFPM method offers the ability to generalize to 3D image/patient registration and to register partial projection data. By applying patient registration directly from tomographic projection data, image reconstruction is not needed in the therapy set-up verification, thus reducing computational time and artefacts. In addition, real time registration is feasible. Registration from partial projection data meets the geometry and dose requirements in many application cases and makes dynamic set-up verification possible in tomotherapy. PMID- 10473213 TI - Application of near-infrared time-resolved spectroscopy to rat liver--a preliminary report for surgical application. AB - The applicability of near-infrared time-resolved spectroscopy to rat liver surgery was investigated. First, the technical reliability in determining the absorption coefficient (mu(a)) and reduced scattering coefficient (mu'(s)) of the liver was checked. Next, boundary effects in determining mu(a) and mu'(s) of the rat liver were examined. Finally, changes in mu(a) and mu'(s) of rat liver with ischaemia were directly measured by TRS. Our TRS system showed that the mu(a) value held a linear correlation with the ink concentration in a lipid emulsion until mu(a) reached 1.2 cm(-1), while the mu'(s) was fairly independent. The mu(a) values of blood-free rat liver and blood-containing rat liver at 780 nm were observed to be 0.43 cm(-1) and 0.67 cm(-1) by using the matching method, indicating that TRS is reliable in determining mu(a) and mu'(s) of the liver. Possible errors in mu(a) and mu'(s) determination due to the boundary effects of the rat liver were as small as 7%, when the mu(a) value was as high as observed for the liver. The oxygen saturation of haemoglobin (SO2) was changed from 64.9% to 8.0%, and the haemoglobin content (THB) from 189.1 microM to 131.6 microM by ischaemia. Mu'(s) dynamically changed in the range 7.06 cm(-1) to 11.36 cm(-1). We conclude that time-resolved measurement is applicable in the high-mu(a) region observed in the liver, and can give quantitative estimations of SO2 and THB in the liver. PMID- 10473214 TI - Feasibility of reading LiF thermoluminescent dosimeters by electron spin resonance. AB - Lithium fluoride is a commonly used solid state dosimeter. During irradiation, electrons and holes become trapped in crystal imperfections; thermoluminescence dosimetry measures their thermally induced recombination. Electron paramagnetic resonance (EPR) spectroscopy can be used to measure the resonant absorption of microwaves by the unpaired electrons trapped in LiF. In an effort to extend the use of LiF dosimeters to smaller sizes and to the harsh environments encountered in internal dosimetry, EPR was evaluated as an alternative technique to read the radiation dose delivered to TLD-100 dosimeters. TLD-100 rods were irradiated with a 60Co source to doses of 10 Gy to 100 Gy. A radiation-induced signal (with a g value of 2.002) could be detected only at liquid nitrogen temperatures at doses above 20 Gy. The EPR spectrum of irradiated LiF contains three components, one of which correlates positively with dose. However, the low sensitivity of the technique, and difficulty in interpreting the EPR spectrum from polycrystalline dosimeters, preclude its use as a dosimetry technique. PMID- 10473215 TI - Comparison of the spectral biologically effective solar ultraviolet in adjacent tree shade and sun. AB - The solar spectral UVR irradiances in tree shade and sunlight have been measured in a sub-tropical southern hemisphere summer. The spectral data allowed the UVB and UVA irradiances and the biologically effective irradiances to be calculated for different harmful biological processes to human skin and eyes. The average of the ratio of the UVA to UVB irradiances was lower by 26% in the shade compared with the same ratio in the sun. The spectral shade ratio calculated as the ratio of the spectral biologically effective irradiances in the shade to those in the adjacent sun decreased with increasing wavelength for all of the trees. The decrease in the shade ratio was approximately 42% at 400 nm compared with the shade ratio at 300 nm. Despite the UVR protection provided by tree shade, the erythemal UVR exposure received in 1 h in the tree shade exceeded the occupational limit for UVR exposure. PMID- 10473216 TI - Linear transformations of data space in MEG. AB - Magnetoencephalography (MEG) is a method which allows the non-invasive measurement of the minute magnetic field which is generated by ion currents in the brain. Due to the complex sensitivity profile of the sensors, the measured data are a non-trivial representation of the currents where information specific to local generators is distributed across many channels and each channel contains a mixture of contributions from many such generators. We propose a framework which generates a new representation of the data through a linear transformation which is designed so that some desired property is optimized in one or more new virtual channel(s). First figures of merit are suggested to describe the relation between the measured data and the underlying currents. Within this context the new framework is established by first showing how the transformation matrix itself is designed and then by its application to real and simulated data. The results demonstrate that the proposed linear transformations of data space provide a computationally efficient tool for analysis and a very much needed dimensional reduction of the data. PMID- 10473217 TI - On the possibility of determining an effective energy spectrum of clinical electron beams from percentage depth dose (PDD) data of broad beams. AB - Experiments have already shown that obvious differences exist between the dose distribution of electron beams of a clinical accelerator in a water phantom and the dose distribution of monoenergetic electrons of nominal energy of the clinical accelerator in water, because the electron beams which reach the water surface travelling through the collimation system of the accelerator are no longer monoenergetic. It is evident that, while calculating precisely the dose distribution of any incident electron beams, the energy spectrum of the incident electron beam must be taken into consideration. In this note we shall present a method for determining an effective energy spectrum of clinical electron beams from PDD data (percentage depth dose data). It is well known that there is an integral equation of the first kind which links the energy spectrum of an incident electron beam with PDD through the dose distribution of monoenergetic electrons in the medium, as a kernel function in the integral equation. In this note, the integral equation of the first kind will be solved by using the regularization method. The bipartition model of electron transport will be used to calculate the kernel function, namely the energy deposition due to monoenergetic electron beams in the medium. PMID- 10473218 TI - Efficient SPECT scatter calculation in non-uniform media using correlated Monte Carlo simulation. AB - Accurate simulation of scatter in projection data of single photon emission computed tomography (SPECT) is computationally extremely demanding for activity distribution in non-uniform dense media. This paper suggests how the computation time and memory requirements can be significantly reduced. First the scatter projection of a uniform dense object (P(SDSE)) is calculated using a previously developed accurate and fast method which includes all orders of scatter (slab derived scatter estimation), and then P(SDSE) is transformed towards the desired projection P which is based on the non-uniform object. The transform of P(SDSE) is based on two first-order Compton scatter Monte Carlo (MC) simulated projections. One is based on the uniform object (P(u)) and the other on the object with non-uniformities (P(nu)). P is estimated by P = P(SDSE) P(nu)/P(u). A tremendous decrease in noise in P is achieved by tracking photon paths for P(nu) identical to those which were tracked for the calculation of P(u) and by using analytical rather than stochastic modelling of the collimator. The method was validated by comparing the results with standard MC-simulated scatter projections (P) of 99mTc and 201Tl point sources in a digital thorax phantom. After correction, excellent agreement was obtained between P and P. The total computation time required to calculate an accurate scatter projection of an extended distribution in a thorax phantom on a PC is a only few tens of seconds per projection, which makes the method attractive for application in accurate scatter correction in clinical SPECT. Furthermore, the method removes the need of excessive computer memory involved with previously proposed 3D model-based scatter correction methods. PMID- 10473219 TI - Penicillin-resistant Streptococcus pneumoniae: selection by both beta-lactam and non-beta-lactam antibiotics. PMID- 10473220 TI - Antimicrobial therapy for Chlamydia pneumoniae: its potential role in atherosclerosis and asthma. PMID- 10473221 TI - Over-the-counter antibiotics. PMID- 10473222 TI - The 1998 Garrod lecture. Current and future antifungal therapy: new targets for antifungal agents. AB - Invasive fungal infections are a major problem in immunocompromised patients. The recent expansion of antifungal drug research has occurred because there is a critical need for new antifungal agents to treat these life-threatening invasive infections. The overview of the development of antifungal therapy which is provided herein reflects the increased interest in this very special area of infectious diseases. Although we have newer, less toxic, antifungal agents that are available for clinical use, their clinical efficacy in some invasive fungal infections, such as aspergillosis and fusariosis, is not optimal. Thus, intense efforts in antifungal drug discovery are still needed to develop more promising and effective antifungal agents for use in the clinical arena. PMID- 10473223 TI - Self-medication of antibacterials without prescription (also called 'over-the counter' use). A report of a Working Party of the British Society for Antimicrobial Chemotherapy. AB - The availability of antimicrobial agents for self-medication may increase and could include antibacterial agents for oral or topical use. Wholesale deregulation of antibacterials would be undesirable and likely to encourage misuse of classes of agents currently important in the management of serious infections. Changed regulation from Prescription-Only Medicine (POM) to Pharmacy (P) medicine of selected agents with indications for short-term use in specific minor infections and illness is likely to have advantages to the user. However, safeguards to their use would need to be included in the Patient Information Leaflet (PIL). Agents and indications for self-medication are discussed. Any alteration in licensed status from POM to P will require careful risk-benefit assessment, including the likely impact on bacterial resistance. Safety issues also include concerns relating to age of the user, pregnancy, underlying disease and the potential for drug interactions. The importance of appropriate information with the PIL is emphasized, as is the role of the pharmacist, while ways of improving adverse event notification and monitoring are discussed. The paucity of good denominator-controlled data on the prevalence of in-vitro resistance is highlighted, and recommendations for improving the situation are made. There are currently no levels of resistance accepted by regulatory bodies on which to base a licensing decision, be it for granting a product licence, renewal of a licence or a change in licensed status from POM to P. Due consideration should be given to: the validation of user-defined indications in comparison with those medically defined; the enhancement of pharmacy advice in the purchase of such agents; improved safety monitoring; the establishment of systematic surveillance of susceptibility data. PMID- 10473224 TI - In-vitro and in-vivo antibacterial activity of BI 397, a new semi-synthetic glycopeptide antibiotic. AB - BI 397 (formerly A-A-1) is a semisynthetic derivative of the teicoplanin-like glycopeptide A40926. It was more active in vitro against staphylococci (including some teicoplanin-resistant strains) than teicoplanin and vancomycin. Against streptococci (including penicillin-resistant strains) BI 397 has activity comparable with that of teicoplanin and better than vancomycin. BI 397, when administered to rats by the i.v. route, gives high and long lasting blood levels. It shows excellent activity in models of acute septicaemia in immunocompetent and neutropenic mice. In a rat staphylococcal endocarditis model it is as effective as teicoplanin and vancomycin at reducing bacterial loads in the heart, but at lower dosages and with a reduced number of daily treatments compared with the two glycopeptide controls. BI 397 is highly efficacious in clearing penicillin susceptible and -resistant pneumococci from lungs of immunocompetent and neutropenic rats. The data from these studies show that BI 397 combines an excellent in-vitro antibacterial activity with favourable pharmacokinetic behaviour resulting in potent in-vivo activity. PMID- 10473225 TI - Intracellular activity of trovafloxacin against Staphylococcus aureus. AB - The effect of trovafloxacin on Staphylococcus aureus ingested by human granulocytes or monocytes was compared with that on S. aureus in cell-free medium. Maximum growth inhibition (E(R,max)) by the antibiotic was 0.530 log10/h for S. aureus within granulocytes, 0.912 log10/h for S. aureus within monocytes, and 1.830-1.916 log10/h for S. aureus in medium. EC50, the concentration at which 50% of the maximum growth inhibition is achieved, did not differ significantly under the conditions investigated. After inhibition of intracellular killing by granulocytes with sodium fluoride, the intracellular antibacterial activity of trovafloxacin was still less than that in medium. A 3.4 times higher concentration was needed to achieve the same effect on phagocytosed S. aureus as in cell-free medium. Trovafloxacin binds more strongly to granulocytes than to monocytes, the respective cellular concentrations being 10 and four times higher than that in medium. In conclusion, the activity of trovafloxacin against S. aureus ingested by human granulocytes or monocytes is less than that against S. aureus in cell-free medium and is not related to the cell-associated concentration. Intracellular conditions are not favourable for the antibacterial activity of trovafloxacin. PMID- 10473226 TI - Structures of existing and new quinolones and relationship to bactericidal activity against Streptococcus pneumoniae. AB - The in-vitro bactericidal profiles of a number of quinolones against Streptococcus pneumoniae were investigated. Tosufloxacin was found to be the most bactericidal quinolone at the optimum bactericidal:MIC ratio (OBMR), followed by levofloxacin, ciprofloxacin and sparfloxacin, in order of potency. After exposure at the OBMR of each quinolone for 2 h, tosufloxacin showed a post-antibiotic effect (PAE) about 2.3-2.6 times longer than the other quinolones. Compounds with a 2,4-difluorophenyl group at the N-1 position in the quinolone nucleus had the greatest bactericidal activity and PAE. This activity exceeded that found with substitution of the quinolone nucleus at the C-7 position. Although the bactericidal activities of the quinolones correlated well with their PAEs, they were not always consistent with their MICs. These results suggest that bactericidal activity and PAE are governed by factors other than those which determine the MIC values, and a 2,4-difluorophenyl group at the N-1 position in the quinolone nucleus may play an important role in the expression of bactericidal activity and PAE against S. pneumoniae. PMID- 10473227 TI - Inhibition of potassium transport and growth of mycobacteria exposed to clofazimine and B669 is associated with a calcium-independent increase in microbial phospholipase A2 activity. AB - Altered phospholipase A2 (PLA2) activity and its relationship to cation (K+, Ca2+) uptake and growth were investigated in mycobacteria exposed to the riminophenazine antimicrobial agents, clofazimine and B669 (0.15-2.5 mg/L). Microbial PLA2 activity was measured using a radiometric thin-layer chromatography procedure, whereas K+ and Ca2+ transport were measured using 86Rb+ or 42K+ and 45Ca2+, respectively. Short-term exposure (15-30 min) of Mycobacterium aurum A+ or the virulent and avirulent isolates of Mycobacterium tuberculosis H37R to the riminophenazines resulted in dose-related enhancement of microbial PLA2 activity, which was associated with inhibition of K+ influx and growth. Uptake of Ca2+ by mycobacteria was unaffected, or minimally affected, by the riminophenazines at concentrations of < or = 0.6 mg/L, whereas higher concentrations resulted in increased uptake of the cation in the setting of decreased microbial ATP concentrations. The results of kinetic studies using a fixed concentration (2.5 mg/L) of B669 demonstrated that riminophenazine-mediated enhancement of PLA2 activity and inhibition of K+ uptake in mycobacteria are rapid and probably related events that precede, by several minutes, any detectable effects on microbial ATP concentrations and uptake of Ca2+. Inclusion of the extracellular and intracellular Ca2+-chelating agents EGTA (0.2-7.2 g/L) and BAPTA/FURA-2 (0.2-9.5 mg/L), individually or in combination, did not prevent the effects of B669 on mycobacterial PLA2 activity or K+ transport, whereas alpha tocopherol, which neutralizes PLA2 primary hydrolysis products, antagonized the inhibitory effects of the riminophenazines on microbial K+ uptake and growth. These results demonstrate that the antimycobacterial activities of clofazimine and B669 are related to a Ca2+-independent increase in mycobacterial PLA2, leading to interference with microbial K+ transport. PMID- 10473228 TI - Involvement of calcium inhibitable binding to the cell wall in the fungicidal activity of CAN-296. AB - CAN-296 is a heat stable, complex carbohydrate (molecular mass 4300 Da) isolated from the cell wall of the filamentous fungus Mucor rouxii. It possesses potent in vitro fungicidal activity against a wide spectrum of pathogenic yeasts, including azole-resistant isolates of Candida albicans and Candida glabrata. As a preliminary step in the study of the mode of action of this novel antifungal agent, we investigated the effect of various cations on the antifungal activity as well as the binding of CAN-296 to intact cells and cell-wall fractions of C. albicans. The antifungal activity of CAN-296 was inhibited by low concentrations of calcium, magnesium and lithium and by high concentrations of barium, cobalt and manganese, but not by potassium and copper. The calcium-mediated inhibition of the antifungal activity of CAN-296 was readily reversible by the removal of calcium by dialysis, and the fungicidal activity of the inhibited compound was fully restored. The uptake/binding of CAN-296 to intact cells and to the cell wall fraction of C. albicans was time and concentration dependent. Maximum uptake/binding was obtained at 5 mg/L within 60 min and was associated with the aggregation of intact cells. Washing intact cells and the cell-wall fraction preincubated with radiolabelled CAN-296 with 150-fold excess of unlabelled compound failed to remove CAN-296 associated with the intact cells and the cell wall fraction, suggesting that the binding of CAN-296 to C. albicans is tight. The uptake/binding of CAN-296 and the drug-mediated aggregation of intact cells were inhibited by calcium in a concentration-dependent manner. The fact that CAN 296 is a fungicidal agent that binds to intact cells and the cell-wall fraction of C. albicans very tightly, together with the observation that calcium was able to inhibit the fungicidal activity as well as the uptake/binding of CAN-296, suggests that the mode of action of this novel antifungal agent may involve interaction with the cell wall of C. albicans. PMID- 10473229 TI - In-vitro activity of dicationic aromatic compounds and fluconazole against Cryptococcus neoformans and Candida spp. AB - We investigated the in-vitro activity of three selected dicationic aromatic compounds for nine clinical isolates of Cryptococcus neoformans and 93 clinical isolates of Candida spp., representing 12 different species, using a broth macrodilution method following NCCLS recommendations. All the clinical isolates were also tested for fluconazole susceptibility. The in-vitro data demonstrate that compounds 39 and 57 have excellent in-vitro activity for all tested strains (MIC 0.19-1.56 mg/L) except Candida pelliculosa. Moreover, compound 39 showed excellent in-vitro fungicidal activity against Candida krusei, Candida glabrata, Candida lusitaniae and Cryptococcus neoformans with MFCs in the range 0.39-6.25 mg/L. Both compounds 39 and 57 showed excellent in-vitro activity against fluconazole-resistant Candida albicans isolates, including a C. albicans strain that contains all known fluconazole-resistant mechanisms. Comparing MIC data from compounds 21, 39 and 57 with fluconazole, we found a statistically significant difference only with compound 39 (P = 0.043). However, comparing MFC data from compounds 21, 39 and 57 with fluconazole, we found statistically significant differences with all three compounds (P < 0.00001). These data indicate the potential antifungal breadth of two bis-benzimidazoles (compounds 39 and 57) as antifungal agents against yeasts. If it can be determined that compounds 39 and 57 are effective and non-toxic in vivo, the prospect of these compounds as clinically useful antifungal agents will be enhanced. PMID- 10473230 TI - The antibacterial efficacy of trovafloxacin against an experimental infection with Listeria monocytogenes in hydrocortisone-treated mice. AB - The efficacy of trovafloxacin in treating Listeria monocytogenes infections in glucocorticosteroid-treated mice was compared with the efficacy of amoxycillin. Swiss mice were treated with daily injections of 2.5 mg hydrocortisone s.c. and then infected i.v. with 1 x 10(7) cfu of L. monocytogenes. Untreated, this level of infection resulted in 100% mortality between day 3 and day 5 after infection. Both s.c. trovafloxacin and amoxycillin were effective in reducing the number of viable L. monocytogenes in the liver and spleen. Although the MIC of amoxycillin for this isolate of L. monocytogenes was lower than that of trovafloxacin (0.063 mg/L versus 0.5 mg/L, respectively), trovafloxacin was more efficacious in vivo after a single dose in the dose range between 12.5 and 100 mg/kg than was amoxycillin. After treatment with trovafloxacin at 100 mg/kg bodyweight od for 3 days, a mean log10 cfu of 1.58 and 2.52 L. monocytogenes could be recovered from the spleens and livers, respectively, whereas after treatment with amoxycillin at 100 mg/kg bodyweight every 8 h for 3 days, the mean 1og10 cfu values were 2.36 and 2.02, respectively. These differences were statistically not significant. Results of the present study show that the antibacterial efficacy of trovafloxacin against L. monocytogenes in our animal model is equivalent to that of amoxycillin. PMID- 10473231 TI - Population pharmacokinetics of amikacin in patients with haematological malignancies. AB - The aim of this study was to analyse the pharmacokinetic behaviour of amikacin in patients with haematological malignancies using a mixed-effect model and sparse data collected during routine clinical care. The patient population comprised 207 haematology patients divided into two groups: one for computing the population model (n = 134) and the other for validation (n = 73). A one-compartment model was used and the following covariates were tested for their influence on clearance and volume of distribution: age, gender, weight, parenteral nutrition, creatinine clearance, stage of antineoplastic treatment (induction, consolidation, intensification), number of weeks postchemotherapy, clinical diagnosis, Eastern Cooperative Oncology Group score, neutropenia, hypoalbuminaemia, concomitant medication (vancomycin and/or amphotericin B), overhydration, and autologous or allogenic bone marrow transplant. The nonlinear mixed-effect model (NONMEM) was used to assess the population pharmacokinetic model of amikacin in these patients. Apart from bodyweight and renal function, acute myeloblastic leukaemia and hypoalbuminaemia proved to be the most important covariates explaining the interindividual variability in amikacin pharmacokinetics in patients with haematological malignancies. The predictive performance of this population model for amikacin serum concentrations seems suitable for clinical purposes. PMID- 10473232 TI - Treatment of typhoid fever with azithromycin versus chloramphenicol in a randomized multicentre trial in India. AB - To compare the clinical and bacteriological efficacies of azithromycin and chloramphenicol for treatment of typhoid fever, 77 bacteriologically evaluable adults, with blood cultures positive for Salmonella typhi or Salmonella paratyphi A susceptible to their assigned drugs, were entered into a randomized open trial at four hospitals in India. Forty-two patients were randomized to receive azithromycin 500 mg p.o. od for 7 days and 35 to receive chloramphenicol 2-3 g p.o. od in four divided doses for 14 days. Thirty-seven patients (88%) in the azithromycin group responded with clinical cure or improvement within 8 days and 30 patients (86%) in the chloramphenicol group responded with cure or improvement. By day 14 after the start of treatment, all patients treated with azithromycin and all except two of the patients treated with chloramphenicol (94%) were cured or improved. Blood cultures repeated on day 8 after start of therapy showed eradication of organisms in 100% of patients in the azithromycin group and 94% of patients in the chloramphenicol group. By day 14 the eradication rate in the chloramphenicol group had increased to 97%. Stool cultures on days 21 and 35 after start of treatment showed no prolonged faecal carriage of Salmonella spp. in either group. These results indicate that azithromycin given once daily for 7 days was effective therapy for typhoid fever in a region endemic with chloramphenicol-resistant S. typhi infection and was equivalent in effectiveness to chloramphenicol given to patients with chloramphenicol-susceptible infections. PMID- 10473233 TI - The efficacy and safety of quinupristin/dalfopristin for the treatment of infections caused by vancomycin-resistant Enterococcus faecium. Synercid Emergency-Use Study Group. AB - A progressive increase in the incidence of vancomycin resistance in strains of Enterococcus faecium (VREF) has severely constrained treatment options for patients with infection caused by this emerging pathogen. Quinupristin/dalfopristin (Synercid), the first injectable streptogramin antibiotic, is active in vitro against VREF, with an MIC90 of 1.0 mg/L. We studied the clinical efficacy and safety of quinupristin/dalfopristin in the treatment of VREF infection. Two prospective studies were conducted simultaneously. The first enrolled only patients with VREF infection; the second included patients with infection caused by other gram-positive bacterial pathogens in addition to VREF. Patients were enrolled if they had signs and symptoms of active infection and no appropriate alternative antibiotic therapy. The recommended treatment regimen of quinupristin/dalfopristin was 7.5 mg/kg i.v. every 8 h for a duration judged appropriate by the investigator. A total of 396 patients with VREF infection were enrolled. The most frequent indications for treatment included intra-abdominal infection, bacteraemia of unknown origin, urinary tract infection, catheter-related bacteraemia, and skin and skin structure infection. This patient population had a high prevalence of severe underlying illness, including a history of diabetes mellitus, transplantation, mechanical ventilation, dialysis, chronic liver disease with cirrhosis and oncological disorders. The mean (+/- S.D.) duration of treatment was 14.5 +/- 10.7 days (range: 1-108). The majority of patients (82.1%) were treated every 8 h, as assessed on day 2 of treatment, while 15.9% were treated every 12 h. The clinical success rate was 73.6% [142/193 clinically evaluable patients; 95% confidence interval (CI): 67.4%, 79.8%], the bacteriological success rate 70.5% (110/156 bacteriologically evaluable patients; 95% CI: 63.4%, 77.7%) and the overall success (both clinical and bacteriological success) rate 65.8% (102/156 bacteriologically evaluable patients; 95% CI: 57.9%, 72.9%). VREF bacteraemia at entry, mechanical ventilation and laparotomy were associated with a worse outcome. Quinupristin/dalfopristin was generally well tolerated. The most common systemic adverse events related to treatment were arthralgias (9.1%) and myalgias (6.6%). Related laboratory abnormalities were infrequent. In these severely ill patients with VREF infection and no other clinically appropriate therapeutic alternatives, quinupristin/dalfopristin demonstrated substantial efficacy and a good nervous system, cardiovascular, gastrointestinal, renal and hepatic tolerability. PMID- 10473235 TI - Endothelial cell compatibility of clindamycin, gentamicin, ceftriaxone and teicoplanin in Bier's arterial arrest. AB - In patients with infected diabetic foot lesions, and gangrenous, peripheral, occlusive arterial disease, it is important to achieve high concentrations of antibiotics in the tissues, as the extent of amputation is often influenced by the presence of infection. Local transvenous pressure injection of antibiotics, in Bier's arterial arrest, allows high local tissue concentrations to be attained in the extremities. Information on the endothelial compatibility of antibiotics in high concentrations combined with the effect of reperfusion injury following tissue hypoxia is lacking. To evaluate the effect of clindamycin, gentamicin, ceftriaxone and teicoplanin injected in Bier's arterial arrest, on endothelial cells, an in-vitro model using human umbilical venous endothelial cells (HUVEC) has been devised. The intracellular levels of purine nucleotides, reflecting DNA/RNA synthesis, energy production and signal transduction of these cells were measured by means of high-performance liquid chromatography. Incubation of cells with 10 mg/mL clindamycin, gentamicin, ceftriaxone and teicoplanin for 20 min resulted in no significant decline of intracellular purines. Levels of purines obtained after exposure of the cells to 0.1 mmol/L hydrogen peroxide (H2O2), to simulate reperfusion injury, were not significantly different from those obtained from cells allowed to recover after antibiotic exposure. These findings indicate that the infusion of high doses of antibiotics, during Bier's arterial arrest, is compatible with maintenance of endothelial cell function, even in the presence of increased free radical activity, provided the exposure is limited to 20 min. PMID- 10473234 TI - Treatment of hospitalized patients with complicated gram-positive skin and skin structure infections: two randomized, multicentre studies of quinupristin/dalfopristin versus cefazolin, oxacillin or vancomycin. Synercid Skin and Skin Structure Infection Group. AB - Quinupristin/dalfopristin (Synercid), the first injectable streptogramin antibiotic available for the treatment of complicated gram-positive skin and skin structure infections, was compared with standard comparators (cefazolin, oxacillin or vancomycin) in one USA and one international trial. These two randomized, open-label trials of virtually identical design enrolled a total of 893 patients (450 quinupristin/dalfopristin, 443 comparator). The majority of patients had erysipelas, traumatic wound infection or clean surgical wound infection. Staphylococcus aureus was the most frequently isolated pathogen in both treatment groups and polymicrobial infection was more common in the quinupristin/dalfopristin group than in the comparator group. The clinical success rate (cure plus improvement) in the clinically evaluable population was equivalent between the two treatment groups (68.2% quinupristin/dalfopristin, 70.7% comparator; 95% CI, -10.1, 5.1) despite a shorter mean duration of treatment for quinupristin/dalfopristin patients. In the bacteriologically evaluable population, by-patient and by-pathogen bacteriological eradication rates were somewhat lower for quinupristin/dalfopristin (65.8% and 66.6%, respectively) than for the comparator regimens (72.7% and 77.7%, respectively). The lower bacteriological response rates in the quinupristin/dalfopristin group were, in part, due to a higher rate of polymicrobial infections and a higher incidence of patients classified as clinical failure, a category which included premature discontinuation of treatment because of local venous adverse events. The bacteriological eradication rate for quinupristin/dalfopristin was higher in monomicrobial infections than in polymicrobial infections (72.6% versus 63.3%, respectively), whereas the corresponding rate for the comparator regimens was lower for monomicrobial infections than polymicrobial infections (70.8% versus 83.1%). This finding was not unexpected, since the spectrum of quinupristin/dalfopristin is focused on gram-positive pathogens and additional antibiotics to treat gram-negative bacteria were not required per protocol. The systemic tolerability of both treatment regimens was qualitatively similar. A higher rate of drug-related venous adverse events was reported for quinupristin/dalfopristin (66.2%) than for the comparator regimen (28.4%). Premature discontinuation of study drug was primarily due to adverse clinical events for quinupristin/dalfopristin (19.1%), whereas the most common reason for discontinuation among those receiving the comparator regimens was treatment failure (11.5%). Quinupristin/dalfopristin is an effective alternative for the treatment of hospitalized patients with complicated skin and skin structure infections due to quinupristin/ dalfopristin-susceptible gram-positive organisms, including methicillin- and erythromycin-resistant S. aureus. PMID- 10473236 TI - Bactericidal action of oral ampicillin/sulbactam against Mycobacterium leprae. AB - We reported previously that an injectable form of ampicillin/sulbactam, Unasyn, was bactericidal to Mycobacterium leprae multiplying in mouse foot pads. In this study, we examined the effect of an orally active form of ampicillin/sulbactam, Sultamicillin, on the growth of M. leprae in mice. Three concentrations of the drug, mixed with the feed, were administered from the start until the mice were killed at 6 months; 0.01% of the drug inhibited bacterial growth by 54%, 0.10% by 74% and 0.20% by 93%. To test whether oral ampicillin/sulbactam was bactericidal, 0.50% of the drug, mixed with the feed, was administered to experimentally infected mice for 3 months during the logarithmic phase of bacterial growth, and then discontinued; multiplication of the bacilli was monitored monthly for the next 8 months. The results showed that orally active ampicillin/sulbactam is bactericidal to M. leprae. PMID- 10473237 TI - In-vitro comparative activity of UR-9825, itraconazole and fluconazole against clinical isolates of Candida spp. AB - UR-9825 is a new broad-spectrum triazole antifungal agent with a good pharmacokinetic profile and excellent bioavailability. It shows high in-vitro activity and efficacy in models of systemic candidosis in rats and rabbits, comparing favourably with fluconazole. The purpose of this study was to evaluate the in-vitro activity of UR-9825 and to compare it with that of fluconazole and itraconazole against 283 clinical isolates of Candida spp. UR-9825 was more potent against Candida spp. than both fluconazole and itraconazole, even against some Candida albicans and Candida krusei isolates with decreased susceptibility to fluconazole (MIC 16 mg/L). PMID- 10473238 TI - Ceftriaxone versus vancomycin prophylaxis in cardiovascular surgery. AB - The efficacy of antibiotic prophylaxis in cardiac surgery was compared between 97 patients receiving a single 2 g dosage of ceftriaxone and 103 receiving 500 mg of vancomycin i.v. every 6 h for 48 h. The overall infection rate was 13.4% in the ceftriaxone and 10.7% in the vancomycin group. Four (4%) wound infections, including one mediastinitis, occurred in the ceftriaxone group and five (5%) in the vancomycin group, with no statistically significant difference. The findings of this study support the adequacy of a simple single dose of ceftriaxone prophylaxis in cardiac surgery, at least in hospitals with low incidence of vancomycin-resistant staphylococcal infections. PMID- 10473239 TI - Personalized antifungal susceptibility testing. PMID- 10473240 TI - Setting the standard for determining the in-vitro susceptibility of Helicobacter pylori to metronidazole. PMID- 10473241 TI - Errors associated with determining the susceptibilities of staphylococci to trimethoprim by the Vitek GPS-AK card. PMID- 10473242 TI - Mycobacterium tuberculosis rpoB gene DNA sequencing: implications for detection of rifamycin resistance. PMID- 10473243 TI - In-vitro activity of gatifloxacin, a novel fluoroquinolone, compared with that of ciprofloxacin against Legionella spp. PMID- 10473244 TI - In-vitro activity of polyhexamethylene biguanide (PHMB) against fungal isolates associated with infective keratitis. PMID- 10473245 TI - Antimicrobial resistance levels of enterobacteria isolated from minced meat. PMID- 10473246 TI - Continuing high prevalence of methicillin resistance amongst Staphylococcus aureus blood culture isolates. PMID- 10473247 TI - Determination of sparfloxacin and its degradation products by HPLC-PDA. PMID- 10473248 TI - Monoaminergic-cholinergic interactions in the primate basal forebrain. AB - Anatomical studies in the rat have shown that the cholinergic cells of the nucleus basalis receive synapses from monoamine axons, but similar evidence is lacking in primates. We used single- and double-labeling immunocytochemistry to visualize monoamine axons and their relationship with the cholinergic cells of the basal forebrain of the monkey. Norepinephrine axons, labeled with dopamine beta-hydroxylase antibodies, formed a bed of fine varicose axons that co distributed with the cholinergic cells. Tyrosine hydroxylase-immunoreactive axons, presumed to be mainly dopaminergic, were 10-20 times more abundant than dopamine-beta-hydroxylase axons throughout the basal forebrain, except in the medial septal area, where their density was lower. Serotonin-immunoreactive axons formed a dense axon plexus throughout the basal forebrain. Double-labeling light microscopy demonstrated that each of the three types of monoamine axons formed frequent direct contacts with the cholinergic cells. Electron microscopy showed that the noradrenergic and the putative dopaminergic axons synapsed on the cholinergic cells. In the human brain, immunolabeling with antibodies to dopamine beta-hydroxylase, tyrosine hydroxylase and tryptophan hydroxylase (for serotonin axons) showed axon densities in the nucleus basalis comparable to those of the monkey brain. The data demonstrate that all three of these monoamine systems innervate the cholinergic and possibly also the non-cholinergic cells of the nucleus basalis, and therefore affect the release of acetylcholine in the cerebral cortex. PMID- 10473249 TI - Serotonin-2 receptor stimulation normalizes striatal preprotachykinin messenger RNA in an animal model of Parkinson's disease. AB - Dopamine and serotonin neurotransmission regulate striatal preprotachykinin messenger RNA levels. In the present study, we investigated serotonin 2A/2C receptor-mediated regulation of preprotachykinin messenger RNA expression in the rat striatum after adult dopamine depletion produced with 6-hydroxydopamine. Significant reductions (46-61% of control values) in preprotachykinin messenger RNA levels were detected by in situ hybridization in rostral, central and caudal regions of the striatum after >85% dopamine depletion. Repeated administration of the specific serotonin2A/2C receptor agonist, (+/-)-2,5-dimethoxy-4 iodoamphetamine hydrobromide, to dopamine-depleted rats completely reversed the reduction in preprotachykinin messenger RNA levels in rostral, central and dorsal caudal striatal regions. In unlesioned (vehicle-injected) control animals, repeated administration of (+/-)-2,5-dimethoxy-4-iodoamphetamine hydrobromide did not affect preprotachykinin messenger RNA expression in rostral, central and ventral-caudal striatal regions, but decreased preprotachykinin messenger RNA levels in the dorsal-caudal striatal subregion. In addition, serotonin turnover in the dopamine-depleted rostral striatum was significantly increased by 35-45% which is consistent with serotonin hyperinnervation after 6-hydroxydopamine lesions. These data show that the decrease in striatal preprotachykinin messenger RNA after dopamine depletion can be normalized with repeated serotonin2A/2C receptor stimulation. We hypothesize that this serotonin2A/2C receptor regulation of preprotachykinin messenger RNA expression after 6-hydroxydopamine is a consequence of serotonin hyperinnervation, which may include increased striatal serotonin2A/2C receptors, induced by dopamine depletion. We also propose that the serotonin system could be pharmacologically targeted to restore the direct striatal tachykinin pathway in Parkinson's disease. PMID- 10473250 TI - Early effects of intrastriatal injections of quinolinic acid on microtubule associated protein-2 and neuropeptides in rat basal ganglia. AB - The long-term effects of intrastriatal injections of the agonist of N-methyl-D aspartate receptors, quinolinic acid, have been extensively characterized. Much less is known, however, about the early molecular and neurochemical changes which occur within a few hours of the toxin injection. In the present study, we have performed intrastriatal injections of low doses of quinolinic acid which induce DNA damage 10-12 h post-lesion, and selective death of striatal projection neurons two weeks later. We examined the time-course of alterations in the microtubule-associated protein 2, an early marker of cytoskeletal disruption, and enkephalin and substance P, two neuropeptides present in largely distinct subpopulations of striatal efferent neurons projecting to the globus pallidus and entopeduncular nucleus, respectively. Immunoreactivity for microtubule-associated protein 2 was decreased at the periphery of the lesion 10 h after quinolinate injection. Levels of enkephalin messenger RNA were markedly decreased as early as 6 h post-lesion; however, a significant decrease in enkephalin immunoreactivity was not observed in the globus pallidus (external pallidum) until 12 h post injection. Levels of substance P messenger RNA were decreased 12 h post-injection in striatal neurons. However, in contrast to enkephalin immunoreactivity, immunolabeling for substance P was not significantly decreased at this time-point in the internal pallidum, a finding reminiscent of early grades of Huntington's disease. The results reveal the time-course of change in messenger RNA and peptide levels in striatal efferent neurons shortly after an excitotoxic insult. These data have implications for the interpretation of findings in post mortem brain and mouse models of Huntington's disease. PMID- 10473251 TI - Effects of lesions to the glutamatergic afferents to the nucleus accumbens in the modulation of reactivity to spatial and non-spatial novelty in mice. AB - The purpose of this study was to compare the effects of selective lesions of the three main sources of limbic afferents to the nucleus accumbens-fornix, prelimbic cortex and amygdala-with those induced by N-methyl-D-aspartate receptor blockage in this structure, in a non-associative task designed to estimate the ability of rodents to encode spatial and non-spatial relationships between discrete stimuli. The task consists of placing mice in an open field containing five objects and, after three sessions of habituation, examining their reactivity to object displacement (spatial novelty) and object substitution (object novelty). Focal administrations of the competitive N-methyl-D-aspartate antagonist DL-2-amino-5 phosphonopentanoic acid (0.1 microg/side) induced a selective impairment in the ability of mice to react to the spatial change. Lesions to the different structures affect the response of mice to spatial and non-spatial novelty in different ways. In particular, while fornix lesions induced a decrease in re exploration of the displaced objects, prelimbic cortex lesions enhanced the exploration of both displaced and non-displaced objects. Finally, the basolateral amygdala lesions did not induce any impairment in the detection of the displaced objects but decreased the latencies to approach novel objects. It is concluded that N-methyl-D-aspartate receptor blockage in the nucleus accumbens subsumes the effects of the three lesions. Some hypotheses on the role of glutamatergic transmission in the accumbens on information processing are briefly discussed. PMID- 10473252 TI - Pretraining tetanic fimbrial stimulation impairs the expression but not the acquisition of contextual fear conditioning in mice. AB - We recently reported that the pretraining induction of long-term potentiation in the lateral septum by fimbrial tetanic stimulation altered contextual fear conditioning in mice. The aim of the present study was to examine at which stage of fear conditioning (i.e. either acquisition or expression) this impairment takes place. Mice implanted with stimulating electrodes in the fimbria and recording electrodes in the lateral septal were conditioned to acquire fear towards a novel context using a footshock procedure. Twenty-four hours after conditioning, animals were re-exposed to the conditioning environment and the level of freezing behavior served as the measure of conditioned fear. The level of fimbrial-lateral septal synaptic neurotransmission was manipulated using either fimbrial tetanic stimulation (which induced septal long-term potentiation) alone, or followed by fimbrial low-frequency stimulation producing depotentiation of the previously established long-term potentiation. The results showed that (i) septal long-term potentiation induced either prior to acquisition or only prior to retention testing impaired conditioned freezing; and (ii) the impairing effect of pretraining induction of long-term potentiation on conditioned freezing was not only abolished by fimbrial low-frequency stimulation administered prior to retention testing but actually produced enhanced conditioned freezing with respect to controls. These data suggest that the level of fimbrial-lateral septal synaptic neurotransmission may influence the expression, but not the acquisition, of contextual fear conditioning. PMID- 10473253 TI - Involvement of central angiotensin receptors in stress adaptation. AB - The present study examined the effects of acute and chronic neurogenic stressors on the expression of two distinct angiotensin receptors in two stress-related brain nuclei: angiotensin type 1A receptor in the paraventricular nucleus of the hypothalamus and angiotensin type 2 receptor in the nucleus locus coeruleus. Male Wistar rats were divided into four experimental groups. The first two groups were subjected once to an acute 90-min immobilization or air-jet stress session, respectively. The other two groups were subjected to 10 days of daily 90-min immobilization sessions and, on the 11th day, one group was exposed to an additional 90-min immobilization and the other to a single air-jet stress (heterotypic but still neurogenic) session. In each group, rats were perfused before stress (0 min), immediately following stress (90 min) or 150, 180, 270 or 360 min (and 24 h in chronic immobilization) after the beginning of the last stress session. Basal expression of both angiotensin receptor subtype 1A and angiotensin receptor subtype 2 messenger RNA was minimal in non-stressed animals. Acute immobilization as well as air-jet stress induced similar patterns (time course and maximal values) of angiotensin receptor subtype 1A messenger RNA expression in the paraventricular nucleus. Angiotensin receptor subtype 1A messenger RNA expression increased 90-150 min after the beginning of the stress and returned to basal levels by 360 min. Chronic stress immobilization slightly modified the pattern, but not maximal values of angiotensin receptor subtype 1A messenger RNA expression to further immobilization (homotypic) or air-jet stress (heterotypic). Acute immobilization and air-jet stress sessions induced similar locus coeruleus-specific angiotensin receptor subtype 2 messenger RNA expression. This expression increased 90 min following the onset of the stress session and remained elevated for at least 360 min. Chronic immobilization stress increased angiotensin receptor subtype 2 messenger RNA expression to levels comparable to those observed in acute stress conditions. Novel acute exposure to neurogenic stressors did not further increase these levels in either homotypic (immobilization) or in heterotypic (air-jet stress) conditions. These results suggest that central angiotensin receptors are targets of regulation in stress; therefore, stress may modulate angiotensin function in the paraventricular nucleus and locus coeruleus during chronic exposure to neurogenic stressors. PMID- 10473254 TI - Amphetamine acts within the medial basal forebrain to initiate and maintain alert waking. AB - Amphetamine-like stimulants exert well-known arousal-enhancing actions. Surprisingly, little is known concerning the neuroanatomical substrates through which these drugs enhance arousal. Previous work implicates a number of basal forebrain structures in the regulation of behavioral state. The current studies examined the effects of amphetamine infusions made directly within basal forebrain sites on behavioral, electroencephalographic, and electromyographic indices of arousal in anesthetized and unanesthetized rat. In the anesthetized rat, amphetamine elicited prolonged epochs of bilateral electroencephalographic activation when infused unilaterally (3.75 microg/150 nl) into an extended region of the medial basal forebrain, demarcated anteriorally by the anterior portion of the medial septal area (which includes posterior accumbens shell) and posteriorally by the posterior aspect of the preoptic area of the hypothalamus. In the unanesthetized (undisturbed, resting) rat, amphetamine infusions into this region elicited prolonged epochs of alert waking, which at the lowest dose (3.75 microg), qualitatively resembled normal waking. Infusions placed lateral (including within the substantia innominata), anterior (including within the core subregion of the nucleus accumbens), posterior, or dorsal to these structures, as well as directly within the lateral ventricles did not alter electroencephalographic or behavioral measures. These results indicate that a region of the medial basal forebrain, extending from the anterior medial septum/accumbens shell to the posterior preoptic area, is a site within which amphetamine-like stimulants act to enhance behavioral and electroencephalographic measures of arousal. PMID- 10473255 TI - Dopaminergic input is required for increases in serotonin output produced by behavioral activation: an in vivo microdialysis study in rat forebrain. AB - Previous research has demonstrated that pharmacological stimulation of postsynaptic dopamine D2 receptors produces increases in serotonin output. The present study explored whether this relationship also holds under physiological conditions. Accordingly, we examined the effects of D2 receptor blockade or unilateral dopamine depletion on behaviorally induced increases in extracellular serotonin levels in the corpus striatum and prefrontal cortex of freely moving rats using in vivo microdialysis. Extracellular levels of dopamine and serotonin, as well as behavioral activity, were increased by both mild tail pinch and the light-dark transition. Tail pinch-induced increases in serotonin levels (39+/-3% and 53+/-5% in the corpus striatum and prefrontal cortex, respectively), but not the accompanying behavioral changes, were blocked by local application of the D2 receptor antagonist raclopride (10 microM). D2 receptor blockade also disrupted the positive relationship between striatal serotonin levels and behavioral activity of animals across the light-dark transition (r=0.93 without raclopride, r=0.24 in presence of raclopride). Unilateral 6-hydroxydopamine lesion of the nigrostriatal dopaminergic system also abolished increases in striatal serotonin output induced by both tail pinch and light-dark transition. A negative correlation was observed between the degree of striatal dopamine depletion and tail pinch-induced increases in serotonin efflux (r= - 0.88). Thus, both a local blockade of postsynaptic D2 receptors and striatal dopamine depletion prevented increases in serotonin output that normally accompany behavioral activation. These data indicate that the increases in the forebrain serotonin output produced by two distinct physiological/environmental manipulations appear to be largely dependent upon intact local dopaminergic neurotransmission. PMID- 10473256 TI - Fast and slow recovery phases of goldfish behavior after transection of the optic nerve revealed by a computer image processing system. AB - As the goldfish is a common experimental animal for vision research, including psychophysical behavior, it is very important to quantitatively score fish behavior. We have previously developed a computer image processing system which can acquire the positional coordinates of goldfish moving freely in an aquarium and determine turning directions (go straight, right or left turn). In the present study, an algorithm to determine tilting angles of moving goldfish was constructed. We also made histograms for quantifying the interaction between pairs of goldfish (two-point distance). By using these histograms, we estimated the time-course of behavioral regeneration after optic nerve transection in goldfish. Control goldfish showed an equal percentage of right or left turns and maintained an upright position in a dorsoventral axis. When the optic nerve of a goldfish was unilaterally sectioned, the goldfish showed predominant turning and slight tilting toward the intact eye. The abnormal turning and tilting behaviors lasted for 10-14 days and then gradually decreased, returning to control behaviors by one month after the unilateral transection. When the optic nerve of a single goldfish was bilaterally sectioned, it did not show any preferential turning and tilting behavior, which is similar to what was observed in control goldfish. However, the trace maps showed that, after bilateral sectioning, fish preferred to cross the center of the tank, which was unlike control fish. In control pairs, one goldfish chased the other with a fixed small range of two point distances. However, in pairs of goldfish with bilateral transection of the optic nerve, the blind goldfish behaved independently of each other, with a long two-point distance. The long two-point distance of the blind goldfish lasted for at least two months and then slowly returned to control two-point distance by four months after bilateral transection. Such fast and slow recovery in goldfish behaviors evoked after unilateral and bilateral transection of the optic nerve is discussed with respect to reconnection of regenerating optic nerves in the fish central nervous system. This computer image processing system is a useful tool with which we can quickly and easily quantify fish behavior. PMID- 10473257 TI - Immunohistochemical localization of interleukin-1beta, interleukin-1 receptor antagonist and interleukin-1beta converting enzyme/caspase-1 in the rat brain after peripheral administration of kainic acid. AB - The temporal and anatomical distribution of members of the interleukin-1 system in the rat brain following intraperitoneal kainic acid administration was studied in relation to neurodegeneration as detected with in situ end labelling. Kainic acid administration (10 mg/kg, i.p.) resulted in the induced expression of interleukin-1beta, interleukin- receptor antagonist and caspase-1p10 immunoreactivity in areas known to display neuronal and tissue damage upon excitotoxic lesions. The induction of these proteins was transient. Interleukin-1 immunoreactivity appeared at 5 h, and the interleukin-1 receptor antagonist immunoreactive cells were first detected at 12 h, whereas the induction of caspase- 1p10 expression was first detected 24 h after kainic acid injection. Double labelling with the microglial marker Ox42 confirmed that both interleukin 1beta and interleukin-1 receptor antagonist were mainly localized in microglial cells. The regional distribution of in situ end-labelled neurons was similar to the distribution of cells expressing interleukin-1beta and interleukin-1 receptor antagonist, whereas the distribution of caspase-1 was more limited. The in situ end-labelled neurons, were, similarly to the interleukin-1beta-positive cells, first detected at 5 h, which is earlier than the induction of caspase-1. Our results show that the induction of IL-1beta and IL-1 receptor antagonist proteins after kainic acid are closely associated with the temporal as well as the anatomical distribution of in situ end-labelled neurons, whereas the induction of caspase-1 protein exhibited a delayed temporal profile and limited distribution. Since cytokine production occurs in activated microglial cells, the inflammatory component seems to be a strong mediator of this type of excitotoxic damage. The late onset of the caspase-1 expression would seem to indicate that this enzyme has no fundamental role in directly causing neuronal cell death induced by systemic kainic acid. PMID- 10473258 TI - Organization of geniculate and trigeminal ganglion cells innervating single fungiform taste papillae: a study with tetramethylrhodamine dextran amine labeling. AB - Single gustatory nerve fibers branch and innervate several taste buds. In turn, individual taste buds may receive innervation from numerous gustatory nerve fibers. To evaluate the pattern of sensory innervation of fungiform papilla bearing taste buds, we used iontophoretic fluorescent injection to retrogradely label the fibers that innervate single taste papillae in the hamster. For each animal, a single taste papilla was injected through the gemmal pore with 3.3% tetramethylrhodamine dextran amine. Fungiform papillae either at the tongue tip (0.5-1.5 mm from the tip) or more posteriorly (1.5-3.0 mm from the tip) were injected. After one to seven days survival, the geniculate and trigeminal ganglia and the tongue were sectioned and examined for labeled cells and fibers, respectively. Analysis of the number and topographic distribution of geniculate cells innervating single taste papillae, shows that: (i) 15 +/- 4 (S.D.) ganglion cells converge to innervate a single fungiform taste bud; (ii) more ganglion cells innervate anterior- (range: 13-35 cells) than posterior-lying buds (range: five to 12 cells), which, in part, may be related to bud volume (microm3); and (iii) ganglion somata innervating a single taste bud are scattered widely within the geniculate ganglion. Analysis of labeled fibers in the tongue demonstrated that two to eight taste buds located within 2 mm of the injected taste bud share collateral innervation with the injected taste bud. Since all buds with labeled fibers were located in close proximity (within a 2-mm radius), widely dispersed geniculate ganglion cells converge to innervate closely spaced fungiform taste buds. Trigeminal ganglion (mandibular division) cells were also labeled in every case and, as with the geniculate ganglion, a dispersed cell body location and collateralization pattern among papillae were observed. This study shows that iontophoresis of tetramethylrhodamine dextran amine, selectively applied to individual peripheral receptor end-organs, effectively locates sensory ganglion cells in two different ganglia that project to these sites. Moreover, the marker demonstrates collateral branches of sensory afferents associated with the labeled fibers and the nearby receptor areas innervated by these collaterals. The labeling of single or clusters of receptor cells, as well as identified sensory afferents, affords future possibilities for combining this technique with immunocytochemistry to establish the relationships of innervation patterns with neurotransmitters and neurotropic substances within identified cells. PMID- 10473259 TI - Evidence that 3,3',5-triiodothyronine is concentrated in and delivered from the locus coeruleus to its noradrenergic targets via anterograde axonal transport. AB - Recent immunohistochemical studies of rat brain triiodothyronine reveal heaviest localization in locus coeruleus perikarya. The cellular distribution is similar to that observed in concomitant studies of tyrosine hydroxylase immunohistochemistry: heavy clumps of immunoreactive triiodothyronine are distributed within locus coeruleus cytosol and in cell processes, leaving cell nuclei unstained. At the same time, in locus coeruleus targets, cell nuclei as well as surrounding neuropil are prominently triiodothyronine labeled. These observations, combined with diverse evidence linking thyroid hormone with norepinephrine at many levels of physiological and pathophysiological function, led to the hypothesis that the locus coeruleus binds and accumulates triiodothyronine and delivers the hormone via anterograde axonal transport to postsynaptic locus coeruleus targets, where nuclear triiodothyronine receptors are densely concentrated. Furthermore, the hypothesis predicts that destruction of locus coeruleus nerve terminals would interrupt this neural route of triiodothyronine delivery and prevent or reduce triiodothyronine labeling of nuclear receptors in noradrenergic target cells. To test this formulation, we gave the specific locus coeruleus lesioning agent, N-(2-chloroethyl)-N-2 bromobenzylamine hydrochloride (DSP-4), to adult male rats and examined their brains three, five and seven days thereafter by triiodothyronine and, in alternate sections, tyrosine hydroxylase immunohistochemistry. Treatment with DSP 4 resulted in specific and selective reduction in tyrosine hydroxylase and triiodothyronine immunohistochemical labeling in cell nuclei and in nerve cell processes within the neuropil of the hippocampus and cerebral cortex at all time periods examined. The results demonstrate that full occupancy of locus coeruleus target cells by triiodothyronine requires the presence of intact locus coeruleus projections and supports the proposal that, like norepinephrine, triiodothyronine delivery to noradrenergic targets occurs through delivery by locus coeruleus terminals. These findings provide strong support for earlier proposals that triiodothyronine functions as a co-transmitter with norepinephrine in addition to or as part of its genomic role in the cells receiving noradrenergic innervation. PMID- 10473260 TI - High level of adenosine A1 receptor-like immunoreactivity in the CA2/CA3a region of the adult rat hippocampus. AB - We describe the immunocytochemical distribution of adenosine A1 receptors in the rat hippocampus. Adenosine A1 receptor-like immunoreactivity was seen on the cell soma and dendrites of pyramidal cells and the cell soma and proximal part of dendrites of granule cells, but not on glial cells. Developmentally, adenosine A1 receptor-like immunoreactivity was diffuse on postnatal day 7 and increased in intensity in individual cells by day 21. In the CA2/CA3a region, the adult pattern of A1 receptor distribution was established by day 28. In the adult rat hippocampus, rostrocaudal inspection revealed that immunoreactivity in CA2/CA3a was greatest. Confocal microscopy revealed differences in the staining patterns for the adenosine A receptor and synaptophysin, a marker of presynaptic terminals. This result suggests that the adenosine A1 receptor might have postsynaptic physiological functions. Double-labeling of adenosine A1 receptors and anterogradely-labeled fibers from the supramammillary nucleus showed that the fibers from the supramammillary nucleus terminate directly on the cell soma of the A1 receptor-immunopositive neurons in CA2/CA3a and the dentate gyrus. These results indicate that the adenosine A 1 receptor in CA2/CA3a and the dentate gyrus are in a position to regulate hippocampal theta activity and that resultant strong synaptic depression in CA2/CA3a could play a role in regulating the intrinsic signal flow between CA3 and CA1. PMID- 10473261 TI - Cannabinoid CB1 receptors are localized primarily on cholecystokinin-containing GABAergic interneurons in the rat hippocampal formation. AB - Localization of cannabinoid CB 1 receptors on GABAergic interneurons in the rat hippocampal formation was studied by double-labeling immunohistochemistry with confocal microscopy. Virtually all CB1-immunoreactive neurons (95%) are GABAergic. CB 1 fluorescence showed a punctate pattern. In contrast, the GABA fluorescence was distributed homogeneously, suggesting that while CB 1 receptors and GABA exist in the same cells they are not localized in the same subcellular compartments. Although virtually all CB1 neurons were GABAergic, many GABAergic neurons did not contain CB1 receptors. GABAergic interneurons in the hippocampal formation can be further divided into subpopulations with distinct connections and functions, using cell markers such as neuropeptides and calcium binding proteins. CB1 receptors were highly co-localized with cholecystokinin and partially co-localized with calretinin and calbindin, but not with parvalbumin. This suggests that cannabinoids may modulate GABAergic neurotransmission at the synapses on the soma and at synapses on the proximal dendrites of the principal neurons, as well as at synapses on other GABAergic interneurons. PMID- 10473262 TI - Synaptic depression induced by pharmacological activation of metabotropic glutamate receptors in the perirhinal cortex in vitro. AB - The perirhinal cortex is crucially involved in various forms of learning and memory. Decrements in neuronal responsiveness occur in the perirhinal cortex with stimulus repetition during visual recognition performance. However, very little is known concerning the underlying mechanisms of synaptic transmission and plasticity in this cortical region. In this study, we provide evidence demonstrating the presence of functional group I, II and III metabotropic glutamate receptors in the rat perirhinal cortex in vitro. Furthermore, the results demonstrate long-lasting synaptic depression in the perirhinal cortex. Extracellular synaptic responses were recorded from superficial layers of the perirhinal cortex directly below the rhinal sulcus, in response to electrical stimuli delivered in the superficial or intermediate layers to the entorhinal or temporal cortex sides of the rhinal sulcus. Evoked synaptic potentials were depressed during bath perfusion of each of the following: the broad-spectrum metabotropic glutamate receptor agonist (1S,3R)-1-aminocyclopentane-1,3 dicarboxylic acid, the selective group I agonist (R,S)-3,5 dihydroxyphenylglycine, the group II agonist (2S,1'R,2'R,3'R)-(2',3' dicarboxycyclopropyl)glycine and the group III agonist (S)-2-amino-4 phosphonobutanoate. Furthermore, there was a long-lasting depression of synaptic transmission following washout of (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid, (R,S)-3,5-dihydroxyphenylglycine or (2S,1'R,2'R,3'R)-(2',3'-dicarboxy cyclopropyl)glycine. Activation of group III metabotropic glutamate receptors by (S)-2-amino-4-phosphonobutanoate did not result in long-lasting changes in synaptic transmission. Thus, the pharmacological activation of metabotropic glutamate receptors can produce short- or long-term changes in synaptic transmission in the perirhinal cortex. It is possible therefore, that metabotropic glutamate receptors are involved in the decrement in neuronal responsiveness associated with visual recognition in the perirhinal cortex. PMID- 10473263 TI - Spreading depression induces expression of calcium-independent protein kinase C subspecies in ischaemia-sensitive cortical layers: regulation by N-methyl-D aspartate receptors and glucocorticoids. AB - Spreading depression is a wave of sustained depolarization challenging the energy metabolism of the cells without causing irreversible damage. In the ischaemic brain, sreading depression-like depolarization contributes to the evolution of ischaemia to infarction. The depolarization is propagated by activation of N methyl-D-aspartate receptors, but changes in signal transduction downstream of the receptors are not known. Because protein phosphorylation is a general mechanism whereby most cellular processes are regulated, and inhibition of N methyl-D-aspartate receptors or protein kinase C is neuroprotective, the expression of protein kinase C subspecies in spreading depression was examined. Cortical treatment with KCl induced an upregulation of protein kinase Cdelta and zeta messenger RNA at 4 and 8 h, whereas protein kinase Calpha, beta, gamma and epsilon did not show significant changes. The gene induction was the strongest in layers 2 and 3, and was followed by an increased number of protein kinase Cdelta immunoreactive neurons. Protein kinase Cdelta and zeta inductions were inhibited by pretreatment with an N-methyl-D-aspartate receptor antagonist, dizocilpine maleate, which also blocked spreading depression propagation, and with dexamethasone, which acted without blocking the propagation. Quinacrine, a phospholipase A2 inhibitor, reduced only protein kinase C5 induction. In addition, N(G)(-nitro-L-arginine methyl ester, a nitric oxide synthase inhibitor, did not influence protein kinase Cdelta or zeta induction, whereas 6-nitro-7 sulphamoylbenzo[f]quinoxaline-2,3-dione, an alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate/kainate receptor antagonist, and the cyclo-oxygenase inhibitors indomethacin and diclophenac tended to increase gene expression. The data show that cortical spreading depression induces Ca2(+)-independent protein kinase C subspecies delta and zeta, but not Ca(2+)-dependent subspecies, through activation of N-methyl-D-aspartate receptors and phospholipase A2. Even though the signal pathway is similar to the induction described previously in ischaemia for genes implicated in delayed neuronal death, the gene inductions observed here are not necessarily pathogenetic, but may represent a general reaction to metabolic stress. PMID- 10473264 TI - Selective inhibition of inositol 1,4,5-triphosphate-induced Ca2+ release in the CA1 region of the hippocampus in the ischemic gerbil. AB - We examined the effect of ischemia on inositol 1,4,5-trisphosphate receptor induced Ca2+ release by functional and morphological approaches, using the gerbil model after 6-h unilateral occlusion of the common carotid artery. Autoradiographic study revealed that the basal uptake of 45Ca2+ into the endoplasmic reticulum and caffeine-induced 45Ca2+ release from the endoplasmic reticulum were normal in the presence of ATP in each ischemic brain region, whereas inositol 1,4,5-trisphosphate receptor-induced 45Ca2+ release from the endoplasmic reticulum was inhibited only in the CA1 region of the hippocampus on the ischemic side. In moderately ischemic gerbils, electron microscopic study demonstrated aggregation of swollen endoplasmic reticulum in the CA1 region of the hippocampus, so that abundant endoplasmic reticulum assembled in close contact to form endoplasmic reticulum cisternal stacks. In severely ischemic gerbils, immunohistochemical analysis of the hippocampus showed loss of type 1 inositol 1,4,5-trisphosphate receptor protein with preservation of immunoreactivity for type 2 and 3 inositol 1,4,5-trisphosphate receptor proteins, which was confirmed by western blot analysis. Such selective inhibition of inositol 1,4,5-trisphosphate receptor-induced Ca2+ release and the loss of type 1 inositol 1,4,5-trisphosphate receptor in the CA1 region of the hippocampus in cerebral ischemia may be associated with its region-specific vulnerability to ischemia. PMID- 10473265 TI - Anoxia-evoked intracellular pH and Ca2+ concentration changes in cultured postnatal rat hippocampal neurons. AB - The ratiometric indicators 2',7'-bis-(2-carboxyethyl)-5-(and-6) carboxyfluorescein and Fura-2 were employed to examine, respectively, intracellular pH (pHi) and calcium ([Ca2+]i) changes evoked by anoxia in cultured postnatal rat hippocampal neurons at 37 degrees C. Under both HCO3-/CO2- and HEPES-buffered conditions, 3-, 5- or 10-min anoxia induced a triphasic change in pHi consisting of an initial fall in pHi, a subsequent rise in pHi in the continued absence of O2 and, finally, a further rise in pHi upon the return to normoxia, which recovered towards preanoxic steady-state pHi values if the duration of the anoxic insult was < or = 5 min. In parallel experiments performed on sister cultures, anoxia of 3, 5 or 10 min duration evoked rises in [Ca2+]i which, in all cases, commenced after the start of the fall in pHi, reached a peak at or just following the return to normoxia and then declined towards preanoxic resting levels. Removal of external Ca2+ markedly attenuated increases in [Ca2+]i, but failed to affect the pHi changes evoked by 5 min anoxia. The latency from the start of anoxia to the start of the increase in pHi observed during anoxia was increased by perfusion with media containing either 2 mM Na+, 20 mM glucose or 1 microM tetrodotoxin. Because each of these manoeuvres is known to delay the onset and/or attenuate the magnitude of anoxic depolarization, the results suggest that the rise in pHi observed during anoxia may be consequent upon membrane depolarization. This possibility was also suggested by the findings that Zn2+ and Cd2+, known blockers of voltage-dependent proton conductances, reduced the magnitude of the rise in pHi observed during anoxia. Under HCO3-/CO2 free conditions, reduction of external Na+ by substitution with N-methyl-D glucamine (but not Li+) attenuated the magnitude of the postanoxic alkalinization, suggesting that increased Na+/H+ exchange activity contributes to the postanoxic rise in pHi. In support, rates of pHi recovery from internal acid loads imposed following anoxia were increased compared to control values established prior to anoxia in the same neurons. In contrast, rates of pHi recovery from acid loads imposed during anoxia were reduced, suggesting the possibility that Na+/H+ exchange is inhibited during anoxia. We conclude that the steady-state pHi response of cultured rat hippocampal neurons to transient anoxia is independent of changes in [Ca2+]i and is characterized by three phases which are determined, at least in part, by alterations in Na+/H- exchange activity and, possibly, by a proton conductance which is activated during membrane depolarization. PMID- 10473266 TI - Nitric oxide and nitric oxide synthase in the early phase of perinatal asphyxia of the rat. AB - The role of nitric oxide, a compound involved in neurotransmission and regulation of cerebral blood flow, in cerebral ischemia is still not fully elucidated yet. Although well studied in adult systems of cerebral ischemia/hypoxia, information on nitric oxide in perinatal asphyxia is limited and, in particular, no direct evidence for its generation has been provided. We therefore decided to study nitric oxide generation in brain of asphyctic rat pups by biophysical and biochemical methods. We used a simple, non-invasive rat model resembling the clinical situation in perinatal asphyxia: rat pups delivered by Caesarean section were placed into a water bath at 37 degrees C still in patent membranes for various asphyctic periods (up to 20 min). Brain pH, cerebral blood flow, neuronal nitrix oxide synthase messenger RNA (by northern and dot blot analysis), immunoreactive protein (by western blot analysis) and nitric oxide synthase activity were determined; generation of nitric oxide was evaluated directly by electron paramagnetic resonance spectroscopy. Neuronal nitric oxide synthase messenger RNA activity and nitric oxide generation were unaffected, whereas neuronal nitric oxide synthase-immunoreactive protein of 150,000 mol. wt was decreased and of 136,000 mol. wt was increased with the length of the asphyctic period. This is the first report on direct evidence for the generation of nitric oxide in perinatal asphyxia and we demonstrate that nitric oxide production remains unaffected even by 20 min of asphyxia, at a time-point when cerebral blood flow was increased four-fold and severe acidosis was present. However, it was found that levels of immunoreactive neuronal nitric oxide synthase of 136,000 mol. wt were increased paralleling the length of asphyxia. Levels of the 150,000 mol. wt immunoreactive neuronal nitric oxide synthase protein decreased, suggesting a different regulation pattern. Thus, the present biochemical and biophysical results form the basis for further investigations on nitric oxide in perinatal asphyxia. PMID- 10473267 TI - Altered adenylyl cyclase responsiveness subsequent to point mutations of Asp 128 in the third transmembrane domain of the delta-opioid receptor. AB - delta-Opioid receptors belong to the superfamily of G protein-coupled receptors, characterized by seven putative transmembrane domains, and have been shown to interact with a host of effector systems. It has been suggested that the charge on the conserved aspartic acid residue at position 128 in transmembrane domain 3 of the delta-opioid receptor contributes to both the conformation of the receptor binding pocket and the molecular rearrangements which accompany the establishment of high-affinity states of the receptor. In light of this, we used site-directed mutagenesis to determine whether this residue participates in the transmission of signals to adenylyl cyclase, the effector with which opioid receptors have been classically associated. Substitution of this aspartic acid (D128) for the neutral amino acid alanine, or the protonated amino acids lysine and histidine, constitutively couples the receptor to adenylyl cyclase, as evidenced by a curtailed response to forskolin stimulation in transfected cells. In addition, this constitutive activity can be blocked by pretreatment of the transfected cells with pertussis toxin. Interestingly, naloxone blocks this effect in cells expressing the D128A mutant, but acts as an agonist at the D128K mutant. Our findings support the hypothesis that the interaction between agonist and receptor promotes conformational changes that may be mimicked, at least in part, by mutation of the aspartate residue at position 128. Furthermore, these changes appear to be involved not only in receptor activation, but also in the functional discrimination between agonists and antagonists. PMID- 10473268 TI - Differential regulation of brain-derived neurotrophic factor messenger RNA cellular expression in the adult rat visual cortex. AB - In this study, we report a comparative analysis of the distribution of brain derived neurotrophic factor messenger RNA in the binocular primary visual cortex of rats analysed at the end of the critical period for monocular deprivation (postnatal day 35) and during adulthood (postnatal day 90). High-resolution non isotopic in situ hybridization coupled with Nissl staining allowed to determine the relative number of neurons expressing brain-derived neurotrophic factor messenger RNA. In postnatal day 90 rats, the relative number of neurons positive for brain-derived neurotrophic factor messenger RNA significantly decreases in layer II/III with respect to postnatal day 35 animals, being constant in all the other cortical layers. Moreover, we demonstrate that dark rearing for 22 days, starting from postnatal day 90, determines: (i) a decrease of the overall level of brain-derived neurotrophic factor messenger RNA with a consequent reduction of labelling intensity in all cells throughout cortical layers II-VI; (ii) an increase of cell numbers expressing brain-derived neurotrophic factor messenger RNA in layers IV and V; and (iii) a decreased intensity of staining for brain derived neurotrophic factor messenger RNA in dendrites after dark rearing. A re exposure to light for 2 h after the period of darkness almost restores the number of brain-derived neurotrophic factor RNA-positive neurons. We conclude that the maturation of brain-derived neurotrophic factor messenger RNA in neurons of layer II/III goes beyond postnatal days 35-40, which can be considered the end of the critical period [Fagiolini M. et al. (1994) Vis. Res., 34, 709-720]. Moreover, we show that the cellular expression of brain-derived neurotrophic factor messenger RNA is regulated by light in adult rats as well as during development. PMID- 10473269 TI - Differential effects of transforming growth factor-beta(s) and glial cell line derived neurotrophic factor on gene expression of presenilin-1 in human post mitotic neurons and astrocytes. AB - Mutations in the presenilin-1 gene are linked to the majority of early-onset familial Alzheimer's disease cases. We have previously shown that the expression of transforming growth factor-beta is altered in Alzheimer's patients, compared to controls. Here we examine presenilin- expression in human post-mitotic neurons (hNT cells), normal human astrocytes, and human brain tumor cell lines following treatment with three isoforms of transforming growth factor-beta, or glial cell line-derived neurotrophic factor, a member of the transforming growth factor-beta superfamily. As the NT2/D1 teratocarcinoma cell line is treated with retinoic acid to induce differentiation to hNT cells, presenilin-1 messenger RNA expression is dramatically increased. Furthermore, there is a 2-3-fold increase in presenilin-1 messenger RNA expression following treatment of hNT cells with growth factors and similar results are found by Western blotting and with immunohistochemical staining for presenilin-1 protein. However, treatment of normal human astrocytes with cytokines results in minimal changes in presenilin-1 messenger RNA and protein. Interestingly, the expression of presenilin-1 in human U87 MG astrocytoma and human SK-N-SH neuroblastoma cells is only increased when cells are treated with glial cell line-derived neurotrophic factor or transforming growth factor-beta3. These findings suggest that endogenous presenilin-1 gene expression in human neurons can be induced by growth factors present in normal and diseased brain tissue. Cytokines may play a major role in regulating expression of presenilin-1 which may affect its biological actions in physiological and pathological conditions. PMID- 10473270 TI - Control of rat hypothalamic pro-opiomelanocortin neurons by a circadian clock that is entrained by the daily light-off signal. AB - Previous studies have clearly demonstrated that the immediate-early gene, c-fos can regulate, through its protein product Fos, the expression of the pro opiomelanocortin gene. In the present study, immunohistochemistry for Fos and beta-endorphin was used to assess the basal activity of hypothalamic pro opiomelanocortin-producing neurons throughout a 12 h light/12 h dark cycle. Here, we showed that Fos is undetectable in most beta-endorphin neurons from late morning until 30 min after light offset in the evening, whereas Fos is spontaneously expressed in these neurons after 1 h following dark onset. The number of beta-endorphin neurons expressing Fos increases continuously during the first half of the dark phase, is maximal at the middle of this phase and decreases through late night and early morning, reaching a nadir 2-3 h after light onset. Acute shifts of lighting parameters allowed us to demonstrate that the light-off signal per se is neither sufficient nor necessary for Fos expression in beta-endorphin neurons. However, when recurrent, this signal is able to entrain Fos expression after a period of adaptation to the new light/dark schedule. Moreover, an expression of Fos in beta-endorphin neurons persists during subjective night in rat exposed to constant light or constant dark for two to three days. Thus, the occurrence of the daily rhythmic increase in the expression of Fos protein in hypothalamic pro-opiomelanocortin neurons exclusively at (subjective) night suggests that these neurons are, most likely, controlled by a (circadian) nocturnal oscillator. Our data also reveal an interesting property of this oscillator: its entrainment by the daily light-to dark transition signal. PMID- 10473271 TI - Velocity response profiles of collicular neurons: parallel and convergent visual information channels. AB - We have recorded from single neurons in the retinorecipient layers of the superior colliculus of the cat. We distinguished several functionally distinct groups of collicular neurons on the basis of their velocity response profiles to photic stimuli. The first group was constituted by cells responding only to photic stimuli moving at slow-to-moderate velocities across their receptive fields (presumably receiving strong excitatory W-type input but not, or only subthreshold, Y-type input). These cells were recorded throughout the stratum griseum superficiale and stratum opticum and constituted 50% of our sample. The second group of cells exhibited excitatory responses only at moderate and fast velocities (presumably receiving excitatory Y-type but not W-type input). These cells constituted only about 7% of the sample and were located principally in the lower stratum griseum superficiale. The third group of cells was constituted by cells excited over the entire range of velocities tested (1-2000 /s) and presumably received substantial excitatory input from both W- and Y-channels. These cells constituted almost 26% of our sample and were located in the lower stratum griseum superficiale, stratum opticum and the upper part of the stratum griseum intermediale. Overall, cells receiving excitatory Y-type input, i.e. the sum of group two and group three cells, constituted about a third of the sample and their excitatory discharge fields were significantly larger than those of cells receiving only W-type input. A fourth distinct group of collicular neurons was also constituted by cells responding over a wide range of stimulus velocities. These cells were excited by slowly moving stimuli, while fast-moving photic stimuli evoked purely suppressive responses. The excitatory discharge fields of these cells (presumably, indicating the spatial extent of the W-input) were located within much larger inhibitory fields, the extent of which presumably indicates the spatial extent of the Y-input. These low-velocity-excitatory/high velocity-suppressive cells were recorded from the stratum griseum superficiale, stratum opticum and stratum griseum intermediale and constituted about 17% of the sample. The existence of low-velocity-excitatory/high-velocity-suppressive cells in the mammalian colliculus has not been previously reported. Low-velocity excitatory/high-velocity-suppressive cells might play an important role in activating "fixation/orientation" and "saccade" premotor neurons recorded by others in the intermediate and deep collicular layers. Overall, in the majority (57%) of collicular neurons in our sample there was no indication of a convergence of W- and Y-information channels. However, in a substantial minority of collicular cells (about 43% of the sample) there was clear evidence of such convergence and about 40% of these (low-velocity-excitatory/high-velocity suppressive cells) appear to receive excitatory input from the W-channel and inhibitory input from the Y-channel. PMID- 10473272 TI - GABA(B) receptor activation protects GABA(A) receptor from cyclic AMP-dependent down-regulation in rat cerebellar granule cells. AB - Interaction between GABAA and GABA(B) receptors was studied in rat cerebellar granule cells in culture, by the whole-cell patch-clamp approach. Our data show that the GABA(B) agonist (-)baclofen is not able, per se, to significantly change the muscimol-activated chloride current. However, (-)baclofen dose-dependently prevents the reduction of GABA(A) receptor function by forskolin, an activator of adenylate cyclase. The effect of baclofen is mediated by a pertussis toxin sensitive G protein. In fact, in cells treated with pertussis toxin, baclofen and forskolin, the toxin is able to block baclofen action, allowing forskolin to act fully. The protective effect by GABA(B) receptor activation under these circumstances is most probably related to the prevention of cyclic AMP increases after forskolin treatment. In fact, in these neurons cyclic AMP and protein kinase A activation result in a down-regulation of GABA(A) receptor function. On the whole, the data indicate the presence of complex modulation of GABA(A) receptors by GABA(B) receptor types in cerebellum granule cells. PMID- 10473273 TI - Spatial correspondence between tactile projection patterns and the distribution of the antigenic Purkinje cell markers anti-zebrin I and anti-zebrin II in the cerebellar folium crus IIA of the rat. AB - We have compared the band-like distribution of the Purkinje cell-specific polypeptides zebrin I and zebrin II with the spatial organization of tactile projections to crus IIa in the cerebellar hemisphere of the rat. Maps of tactile responses in the granular layer of the cerebellar hemispheres are fractured into discontinuous regions, termed "patches". High-density micromapping was used to identify specific patches and their boundaries within this fractured somatotopic map. In one series of experiments, medial and lateral boundaries of the large central ipsilateral upper lip-related patch were identified and labeled with either Fast Blue or India Ink. Following immunocytochemical processing, the band like distribution of immunostained Purkinje cells (zebrin-positive bands) and the identified patch boundaries were digitized and reconstructed in three dimensions. Comparisons between these two features demonstrate a spatial correspondence between zebrin transitions and the boundaries of the electrophysiologically defined upper lip-related patch. In another series of experiments, we outlined the boundaries or centers of several smaller patches consistently located in the medial portion of the folium. Again, we found a correspondence between the distribution of granule cell layer tactile patches and the zebrin staining pattern. The correspondence between tactile projection patterns and molecular features demonstrated in the present study implies that there is a distinct and largely fixed spatial pattern of organization in the cerebellar hemispheres. We discuss possible causal connections and developmental determinates, as well as the physiological significance of the correspondence between the two features. PMID- 10473274 TI - Neck input modifies the reference frame for coding labyrinthine signals in the cerebellar vermis: a cellular analysis. AB - The activity of 68 neurons, mainly Purkinje cells, was recorded from the cerebellar anterior vermis of decerebrate cats during wobble of the whole animal (at 0.156 Hz, 5 degrees), a mixture of tilt and rotation, leading to stimulation of labyrinth receptors. Most of the neurons (65/68) were affected by both clockwise and counterclockwise rotations. Twenty-four units showing responses of comparable amplitude to these stimuli (narrowly tuned cells) were represented by a single vector (Smax), whose preferred direction corresponded to the direction of stimulation giving rise to the maximal response. The remaining 41 units, however, showed different amplitude responses to these rotations (broadly tuned cells) and were characterized by two spatially and temporally orthogonal vectors (Smax and Smin), suggesting that labyrinthine signals with different spatial and temporal properties converged on these cells. All these units were tested while the body was aligned with the head (control position), as well as after static displacement of the body under a fixed head by 15 degrees and/or 30 degrees around a vertical axis passing through C1-C2, thus leading to stimulation of neck receptors. The orientation component of the response vector of the Purkinje cells to vestibular stimulation changed following body-to-head displacement. Moreover, the amplitude of vector rotation corresponded, on the average, to that of body rotation. Changes in temporal phase, gain and tuning ratio of the responses were also observed. We propose that information from neck receptors regulates the convergence of labyrinthine signals with different spatial and temporal properties on corticocerebellar units. Due to their strict relationship with the motor system, these units may give rise to appropriate responses in the limb musculature, by modifying the spatial organization of the vestibulospinal reflexes according to the requirements of body stability. The cerebellar vermis may thus represent an important structure, where frames of reference can be altered to account for changes in position of trunk, head and neck. PMID- 10473275 TI - Mediation and modulation by eicosanoids of responses of spinal dorsal horn neurons to glutamate and substance P receptor agonists: results with indomethacin in the rat in vivo. AB - In view of the widespread use of non-steroidal anti-inflammatory drugs for treatment of inflammatory pain, we determined the effects of the non-steroidal anti-inflammatory drug, indomethacin, on dorsal horn neurons in the rat spinal cord in vivo. At 2.0-12.0 mg/kg (i.v.), indomethacin depressed the responses of spinal dorsal horn neurons to the effects of iontophoretic application of substance P, N-methyl-D-aspartate, quisqualate and alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionate. As indomethacin inhibits cyclo-oxygenase, these are the first data linking prostanoids and possibly arachidonic acid and other eicosanoids to the effects of substance P and glutamate in the spinal dorsal horn. As responses to iontophoretic application can be assumed to have been postsynaptic and as indomethacin had an effect generalized to all excitatory responses, we suggest a postsynaptic site for cyclo-oxygenase. We also suggest that elements in the cyclo-oxygenase signal transduction pathway may thus mediate at least some of the effects of substance P and glutamate receptor activation. Activation of the cyclo-oxygenase pathway in CNS neurons is Ca2- dependent, and activation of both N-methyl-D-aspartate and substance P receptors increases intracellular Ca2+. This led to the expectation that indomethacin would have a greater effect on responses to N-methyl-D-aspartate than to alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionate, but the reverse was observed. Thus, in addition to a mediator role, we hypothesize that an element(s) of the cyclo-oxygenase pathway may regulate the efficacy of excitation of alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionate receptors and perhaps other membrane-bound receptors. The cyclo-oxygenase signal transduction pathway thus appears to play at least two major roles in regulation of sensory processing in the spinal cord. Therefore, non-steroidal anti-inflammatory drugs, via cyclo-oxygenase inhibition, may have multiple actions in control of spinal sensory mechanisms. PMID- 10473276 TI - Glutamatergic receptors regulate expression, phosphorylation and accumulation of neurofilaments in spinal cord neurons. AB - Glutamatergic regulation of neurofilament expression, phosphorylation and accumulation in cultured spinal cord neurons was studied. At seven days in culture, 0.15% of the neurons were immunoreactive for non-phosphorylated neurofilaments, but essentially no cells immunoreactive for phosphorylated neurofilaments were seen. The number and size of the immunoreactive cells in culture corresponded well to those of rat and human spinal cord neurons in vivo. In spinal cord cultures, sublethal, long-lasting stimulation of alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate or metabotrophic receptors, but not N-methyl-D-aspartate receptors, dose-dependently increased the number of non-phosphorylated neurofilament-immunoreactive cells, which was blocked by nifedipine, an antagonist of voltage-sensitive Ca2+ channels. Stimulation of kainate or all non-N-methyl-D-aspartate receptors decreased the expression of medium-molecular-weight neurofilament messenger RNA. Blockade of AMPA/kainate receptors, but not of N-methyl-D-aspartate receptors, increased the amount of phosphorylated neurofilament protein and the number of phosphorylated neurofilament-immunoreactive cell bodies. The phosphorylated neurofilament immunoreactive cell population was different from the non-phosphorylated neurofilament-immunoreactive neurons, which lost their axonal non-phosphorylated neurofilament immunoreactivity but showed intense cytoplasmic labeling in response to the blockade of AMPA/ kainate receptors. Immunoreactivity for phosphoserine did not change upon glutamate receptor stimulation and blockade. The results show that activation of AMPA/kainate receptors decreases the expression of neurofilament messenger RNA and neurofilament phosphorylation in spinal cord neurons by a mechanism involving active voltage-sensitive Ca2+ channels. Blockade of these receptors seems to disturb axonal neurofilament transport. Because AMPA/kainate receptors mediate chronic glutamatergic death of spinal motor neurons and these receptors have been suggested to be involved in the pathogenesis of amyotrophic lateral sclerosis, the observed alteration in neurofilament phosphorylation and distribution may contribute to the pathogenesis of chronic motor neuron diseases. PMID- 10473277 TI - Glutamate-induced increase of extracellular glutamate through N-methyl-D aspartate receptors in ethanol withdrawal. AB - Ethanol withdrawal is a physiopathological state associated with increased number and function of N-methyl-D-aspartate glutamate receptors. We assessed the effect of N-methyl-D-aspartate receptor stimulation on the extracellular levels of glutamate in vivo by the focal application of N-methyl-D-aspartate in the striatum of dependent rats following withdrawal from chronic treatment with ethanol. In control, chronic sucrose-treated rats, 800 microM N-methyl-D aspartate increased glutamate levels to 268% of baseline values. In ethanol withdrawn animals, 12 h after interruption of the chronic treatment, the application of N-methyl-D-aspartate increased glutamate levels to 598% of baseline values. In ethanol-intoxicated rats N-methyl-D-aspartate was ineffective. Concentration-response curves showed that in ethanol withdrawn animals N-methyl-D-aspartate was five-fold more potent than in controls. In withdrawn animals, the non-competitive N-methyl-D-aspartate receptor antagonist dizocilpine (1.0 mg/kg i.p.) or ethanol (5 g/kg i.g.) markedly reduced the N methyl-D-aspartate-induced increase in glutamate levels. These results are consistent with the up-regulation of N-methyl-D-aspartate receptors by chronic ethanol and add biochemical evidence for the presence of N-methyl-D-aspartate receptors facilitating glutamate release through a positive feedback mechanism. The glutamate-induced, N-methyl-D-aspartate receptor-mediated elevations of extracellular glutamate may constitute a neurochemical substrate for the neuropathological alterations associated with alcoholism. PMID- 10473278 TI - Stabilizing neuromuscular contacts reduces motoneuron death caused by paralysis of muscles in neonatal rats. AB - Transient paralysis of the soleus muscle in neonatal rats leads to permanent muscle weakness, loss of muscle fibres and motoneuron death. Application of leupeptin, an inhibitor of a calcium-activated neutral protease, to the neuromuscular junction is known to enhance the maintenance of neuromuscular contacts during development and axonal sprouting. Here, we show that treatment of soleus muscles with leupeptin as they recover from a period of paralysis rescues motoneurons that would otherwise die. The number of motoneurons to the soleus muscle was established by retrograde labelling with horseradish peroxidase eight to 10 weeks after recovery from paralysis. There were only 38.4 (+/-2.8 S.E.M., n=5) motoneurons innervating the soleus muscle that had been paralysed with alpha bungarotoxin, compared to 58.2 (+/-3.1 S.E.M., n=5) to the control untreated soleus. Thus, the number of motoneurons to the soleus muscle on the alpha bungarotoxin-treated side was 66.9% (+/-6.2% S.E.M., n=5) of the control side. In those animals where paralysis of the soleus muscle was followed three days later by treatment with leupeptin, the number of labelled motoneurons on the treated side of the spinal cord was 61.5 (+/-4.6 S.E.M., n=4) and that on the contralateral untreated control side was 59 (+/-3.8 S.E.M., n=4). This improvement in motoneuron survival in the leupeptin-treated animals is also confirmed by counts of the number of motor units in the soleus muscle obtained by recording muscle tension. In animals that had their soleus muscles paralysed at birth, only 21 (+/-0.7 S.E.M., n=5) motor units were present, compared to 30 motor units in control muscles. When the paralysed soleus muscle was subsequently treated with leupeptin, the number of remaining motor units in the muscle was 29.8 (+/- 1.0 S.E.M., n=5). In addition, the force output of the soleus muscles that had undergone a period of neonatal paralysis was calculated for both the NaCl- and leupeptin-treated animals. The results showed that paralysis at birth results in a reduction in weight and force output of the soleus muscle, which is not improved following treatment with leupeptin. This study shows that application of leupeptin to the soleus muscle after alpha-bungarotoxin-induced paralysis rescues motoneurons to the soleus that would otherwise die. This effect is most likely due to stabilization of their neuromuscularjunctions. PMID- 10473279 TI - Androgen-sensitive preganglionic neurons innervate the male rat pelvic ganglion. AB - In adult male rats many pelvic autonomic ganglion cells change in structure and function after androgen deprivation. In this study we have investigated whether preganglionic neurons in the lumbar and sacral spinal cord that innervate these ganglion cells are also androgen-sensitive. Numerous spinal neurons retrogradely labelled from the pelvic ganglion possessed androgen receptor immunoreactivity and this was diminished by castration or enhanced by additional testosterone exposure. These comprised 27-77% of all preganglionic neurons innervating the pelvic ganglion, depending on the spinal level and whether animals were administered testosterone prior to sacrifice or not. When adult animals were castrated, no change occurred in the soma size or number of primary dendrites in these lumbar or sacral preganglionic neurons. Mean dendrite length was also determined in lumbar preganglionic neurons supplying the pelvic ganglion, but was not affected by castration. However, the total volume of lumbar preganglionic terminal varicosities supplying each noradrenergic pelvic ganglion cell decreased in parallel with the volume of the target neuron. These studies show that many preganglionic autonomic neurons involved in pelvic reflexes are androgen sensitive, but that androgens selectively influence particular neuronal compartments. The prevalence of androgen receptors in these neurons suggests that testosterone may directly influence gene expression of preganglionic neurons. Together these studies suggest that testosterone (or a metabolite) has widespread actions on pelvic reflex circuits during adulthood and that under conditions of diminished circulating androgens a variety of reflex activities may not function optimally. PMID- 10473280 TI - GABA(A) receptor subunit messenger RNA expression in the enteric nervous system of the rat: implications for functional diversity of enteric GABA(A) receptors. AB - GABAergic neurons occur in the myenteric plexus and submucosa and their innervations of the gut, where GABA stimulates motor neurons, and non-neural cells via "central type" GABA(A) receptors. These receptors occur on half of the neurons in the rat intestine. The GABA(A) receptor is a ligand-gated chloride channel constructed from different subunit families (alpha, beta, gamma, delta, epsilon). In rat these exist as subtypes, alpha1-6, beta1-3, gamma1-3 and delta, defining the clinically relevant pharmacological features of GABA(A) receptors. However, the identity, distribution, and abundance of enteric GABA(A) receptor subunits are unknown. To identify and map the regional expression of GABA(A) receptor subunit messenger RNAs in the enteric nervous system, we assayed enteric RNA from the ileum of Sprague-Dawley rats by reverse transcription-polymerase chain reaction for alpha1-6, beta 1-3, gamma1-3, and delta subunit messenger RNAs. Subunit messenger RNA localization, was probed by in situ hybridization. Reverse transcription-polymerase chain reaction analysis of RNA from myenteric and submucosal nerve layers revealed the expression alpha1, alpha3, beta2, beta3, gamma1 and gamma3 subunit messenger RNAs. Little alpha4 and alpha6 and no alpha2, beta1, gamma2 or delta subunit messenger RNA were detected. In situ hybridization revealed that transcripts for alpha1, alpha3, alpha5 and beta2 subunits occur in both myenteric and submucous ganglia. However, beta3 messenger RNA was found only in myenteric plexus. The gamma1 subunit messenger RNA was also restricted to the cells in the myenteric plexus while gamma3 was found in cells of both nerve layers. In this study of the subunit messenger RNA expression profile of GABA(A) receptors within the enteric nerve layers we show an abundant, diverse and widespread distribution that is unique in comparison to the CNS. The distinctive and heterogeneous distribution of enteric GABA(A) subunits may be important in the integration of neural control of gut function. PMID- 10473281 TI - Inhibition of excitatory neurotransmitter-nitric oxide signaling pathway by inhalational anesthetics. AB - Primary cultures of cerebral neurons of Sprague-Dawley rats increased cyclic GMP production in response to the stimulation of excitatory amino acids, including N methyl-D-aspartate, quisqualate, kainate and (+/-)-1-aminocylopentane-trans-1,3 dicarboxylic acid. This increased cyclic GMP production was significantly inhibited by halothane or isoflurane at clinically relevant concentrations (0.5 2%). This inhibition was reversible by treatment with L-arginine, the substrate of nitric oxide synthase. However, the increase of cyclic GMP production stimulated by sodium nitroprusside, an activator of soluble guanylate cyclase, was not inhibited by halothane or isoflurane. Neither halothane nor isoflurane affected the basal cyclic GMP production. Activation of the excitatory amino acid neurotransmitter-stimulated nitric oxide-guanylate cyclase signaling pathway increases intracellular cyclic GMP content in neurons. Our results suggest that halothane or isoflurane inhibited this signaling pathway stimulated by selective agonists of each subtype of receptors for excitatory amino acid neurotransmitters. This inhibition may be involved in mechanisms of anesthesia and analgesia. The site(s) of the inhibition is (are) proximal to the activation of neuronal nitric oxide synthase. PMID- 10473282 TI - Susceptibility to focal and generalized seizures in Wistar rats with genetic absence-like epilepsy. AB - The susceptibility to develop cortically induced focal and generalized seizures was examined in Genetic Absence Epilepsy Rats from Strasbourg (GAERS), an inbred strain of Wistar rats with absence epilepsy. A GABA-withdrawal syndrome induced after suppression of a 2-h intracortical GABA infusion was used as a model of focal epileptogenesis: localized cortical discharges appear at the infusion site within 1 h. GAERS were more prone to develop a GABA-withdrawal syndrome than non epileptic inbred controls and non-selected Wistar rats. After a transient suppression of absence seizures following GABA infusion in GAERS, generalized spike-and-wave discharges and focal spikes were recorded simultaneously in the cortex. GAERS also showed a higher incidence of systemic pentylenetetrazol induced convulsions at the dose of 25 mg/kg. Higher doses had similar convulsant effects in all groups. In conclusion, the results confirm a genetic susceptibility in GAERS and/or resistance in inbred non-epileptic rats to focal and generalized seizures involving the cortex. Rats with absence epilepsy appear to be more prone to seizures elicited by cortical GABA deficiency. PMID- 10473283 TI - Expression profile of the copper homeostasis gene, rAtox1, in the rat brain. AB - In humans the regulation of cellular copper homeostasis is essential for proper organ development and function. A novel cytosolic protein, named Atox 1, was recently identified in yeast that functions in shuttling intracellular mononuclear copper [Cu(I)] to copper-requiring proteins. Atox 1 and its human homolog, hAtox1, are members of an emerging family of proteins termed copper chaperones that are involved in the maintenance of copper homeostasis. Northern blot analysis demonstrates that Atox 1 is widely expressed at varying levels in a variety of rat tissues including brain. Using in situ hybridization histochemistry, we characterized the expression profile for the rat homolog of Atox1 (rAtox1) in the normal adult rat brain. There is widespread expression within the brain that appears to be primarily neuronal. The highest levels of Atox1 message consists of distinct neuronal subtypes that are also characterized by their high levels of metals like copper, iron, and zinc, which include the pyramidal neurons of the cerebral cortex and hippocampus in addition to the neurons of the locus coeruleus. The high levels of a metal chaperone like Atox1 in subsets of neurons that also sequester metals suggests that Atox1 may be important in maintaining the functionality of metal requiring enzymes. A detailed analysis of the restricted expression profile for a novel copper chaperone, rAtox1, is described in the adult rat CNS. Further analysis shows that Atoxl expression is associated with neuronal populations that sequester copper. PMID- 10473284 TI - Cellular distribution of iron in the brain of the Belgrade rat. AB - In this study, we investigated the cellular distribution of iron in the brain of Belgrade rats. These rats have a mutation in Divalent Metal Transporter 1, which has been implicated in iron transport from endosomes. The Belgrade rats have iron positive pyramidal neurons, but these are fewer in number and less intensely stained than in controls. In the white matter, iron is normally present in patches of intensely iron-stained oligodendrocytes and myelin, but there is dramatically less iron staining in the Belgrade rat. Those oligodendrocytes that stained for iron did so strongly and were associated with blood vessels. Astrocytic iron staining was seen in the cerebral cortex for both normal rats and Belgrade rats, but the iron-stained astrocytes were less numerous in the mutants. Iron staining in tanycytes, modified astrocytes coursing from the third ventricle to the hypothalamus, was not affected in the Belgrade rat, but was affected by diet. The results of this study indicate that Divalent Metal Transporter 1 is important to iron transport in the brain. Iron is essential in the brain for basic metabolic processes such as heme formation, neurotransmitter production and ATP synthesis. Excess brain iron is associated with a number of common neurodegenerative diseases. Consequently, elucidating the mechanisms of brain iron delivery is critical for understanding the role of iron in pathological conditions. PMID- 10473285 TI - Migrational analysis of the constitutively proliferating subependyma population in adult mouse forebrain. AB - Initial experiments to evaluate the in vivo fate(s) of constitutively proliferating subependymal cells determined that, following in vivo labeling of this population by infection with a retrovirus containing a beta-galactosidase reporter gene, there was a progressive and eventually complete loss of histochemically beta-galactosidase-positive cells within the lateral ventricle subependyma with increasing survival times of up to 28 days after retroviral infection. Subsequent experiments were designed to ascertain the potential contributions of: (i) the migration of subependymal cells away from the forebrain lateral ventricles; and (ii) the down-regulation of the retroviral reporter gene expression. Retroviral lineage tracing experiments demonstrate that a major in vivo fate for constitutively proliferating subependymal cells is their rostral migration away from the walls of the lateral ventricle to the olfactory bulb. Although down-regulation of retroviral reporter gene expression does not contribute to the loss of detection of beta-galactosidase-labeled cells from the lateral ventricle subependyma, it does result in an underestimation of the absolute number of retrovirally labeled cells in the olfactory bulb at longer survival times. Furthermore, a temporal decrease in the double labeling of beta galactosidase-labeled cells with [3H]thymidine was observed, indicating that only a subpopulation of the migratory subependymal-derived cells continue to actively proliferate en route to the olfactory bulb. These two events may contribute to the lack of a significant increase in the total number of retrovirally labeled subependymal cells during rostral migration. Evidence from separately published studies suggests that cell death is also an important regulator of the size of the constitutively proliferating subependymal population. In summary, in vivo studies utilizing retroviral reporter gene labeling demonstrate that constitutively proliferating subependymal cells born in the lateral ventricle migrate rostrally to the olfactory bulb. Loss of proliferation potential and retroviral reporter gene down-regulation contribute to the lack of any significant increase in the total number of labeled cells recovered in the olfactory bulb. PMID- 10473286 TI - Hyperpolarization-activated inward potassium and calcium-sensitive chloride currents in beating pacemaker insect neurosecretory cells (dorsal unpaired median neurons). AB - Hyperpolarization-activated inward currents were studied in single adult cockroach Periplaneta americana pacemaker neurosecretory cells, identified as dorsal unpaired median neurons using the whole-cell patch-clamp technique. Under current clamp, injection of negative current produced a hyperpolarization of the cell membrane with a sag in the membrane potential toward the resting value. Under voltage clamp, the whole-cell current-voltage relationship exhibited an unexpected biphasic aspect. The global hyperpolarization-activated inward current could be dissociated by means of 4-acetamido-4'-isothiocyanostilbene-2,2' disulfonic acid and tetraethylammonium chloride sensitivity, ionic selectivity, voltage dependence and activation threshold as inward potassium and calcium sensitive chloride currents. The inward potassium current was activated around 80 mV. The reversal potential followed the potassium equilibrium potential when the extracellular potassium concentration was raised. This current was not dependent on the external sodium concentration and was sensitive to 10 mM tetraethylammonium chloride or 5 mM barium chloride. The hyperpolarization activated inward calcium-sensitive chloride current was activated in a range of potential 20 mV more positive than the potassium current. The estimated reversal potential (-71 mV) was very close to the equilibrium potential for chloride ions ( 73 mV). Intracellularly applied 4-acetamido-4'-isothiocyanostilbene-2,2' disulfonic acid, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and external application of 1 mM zinc chloride, calcium-free saline or high concentrations of intracellular 1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetate blocked the inward chloride current. Current-clamp experiments indicated that the inward potassium current accounted for inward rectification of dorsal unpaired median neurons. Our findings report, for the first time in pacemaker neurosecretory cells, the co-existence of two distinct hyperpolarization-activated inward currents which have specialized function in pacemaker activity. PMID- 10473287 TI - The topography, architecture and structure of the enteric nervous system in the jejunum and ileum of cattle. AB - To date, there appear to have been no detailed and clear descriptions of the nerve plexuses and their subdivisions in the intestine of cattle. In this study, the enteric nervous system in the jejunum and ileum of 12 1-y-old calves was examined using neurofilament protein and vasoactive intestinal peptide immunohistochemistry in wholemounts and paraffin sections combined with staining of paraffin and historesin sections with haematoxylin and eosin. The main organisation of the plexuses was similar to that of the pig, horse and man with external and internal submucous plexuses being morphologically distinct, with further subdivisions of the internal submucous plexus into the external and internal subplexuses. However, in contrast to pig, horse and man, the submucous layer was firmly attached to the inner circular muscle layer. The myenteric plexus was well developed with large ganglia, and primary and secondary nerve strands. Its main axis was oriented parallel to the outer longitudinal smooth muscle; large ganglia and primary nerve strands fused to form complex ganglia, and 2 types of tertiary nerve strands were observed. Antibodies to neurofilament proteins and vasoactive intestinal peptide revealed adendritic, pseudouniaxonal or multiaxonal type II neurons only in the myenteric and submucous plexuses. This appears to be the first report of the identification of isolated uniaxonal, multidendritic type IV neurons in the mucous pericryptal plexus. The new information presented here provides further evidence for the existence of anatomical and functional differences between the external and internal submucous plexuses and for supporting the nomenclature proposed earlier. PMID- 10473288 TI - Mu opioid receptors in developing human spinal cord. AB - The distribution of mu opioid receptors was studied in human fetal spinal cords between 12-13 and 24-25 wk gestational ages. Autoradiographic localisation using [3H] DAMGO revealed the presence of mu receptors in the dorsal horn at all age groups with a higher density in the superficial laminae (I-II). A biphasic expression was noted. Receptor density increased in the dorsal horn, including the superficial laminae, between 12-13 and 16-17 wk. This could be associated with a spurt in neurogenesis. The density increased again at 24-25 wk in laminae I-II which resembled the adult pattern of distribution. A dramatic proliferation of cells was noted from the region of the ventricular zone between 16-17 and 24 25 wk. These were considered to be glial cells from their histological features. Mu receptor expression was noted over a large area of the spinal cord including the lateral funiculus at 24-25 wk. This may be due to receptor expression by glial cells. The study presents evidence of mu receptor expression by both neurons and glia during early development of human spinal cord. PMID- 10473289 TI - Increase in liver pigmentation during natural hibernation in some amphibians. AB - The amount/distribution of liver melanin in 3 amphibian species (Rana esculenta, Triturus a. apuanus, Triturus carnifex) was studied during 2 periods of the annual cycle (summer activity-winter hibernation) by light and electron microscopy, image analysis and microspectrofluorometry. The increase in liver pigmentation (melanin content) during winter appeared to be correlated with morphological and functional modifications in the hepatocytes, which at this period were characterised by a decrease in metabolic activity. These findings were interpreted according to the functional role (e.g. phagocytosis, cytotoxic substance inactivation) played by the pigment cell component in the general physiology of the heterothermic vertebrate liver and, in particular, in relation to a compensatory engagement of these cells against hepatocellular hypoactivity during the winter period. PMID- 10473290 TI - Ultrastructural study of the relationship between the morphogenesis of filiform papillae and the keratinisation of the lingual epithelium in the rat. AB - Tongues were removed from rat fetuses on d 16 of gestation (E16) and from newborn (P0) and juvenile rats on d 7 (P7) and d 21 (P21) postnatally for examination by light and transmission electron microscopy. In the fetuses at E16, no rudiments of filiform papillae were visible on the dorsal surface of the tongue. No evidence of keratinisation could be recognised over the entire dorsal lingual epithelium. At P0, rudiments of filiform papillae showed a similar distribution to that seen in the adult, but had a more rounded appearance. The columnar structure of cells in the epithelium, with the different degrees of keratinisation as observed in the mature adult, was indistinct, but a keratinised layer was clearly located at the tip of each filiform papilla. In juveniles at P7, the filiform papillae on the anterior part of the tongue were long and slender, and the anterior and posterior cell columns of the filiform papillae and the interpapillary cell columns were clearly distinguishable. In juveniles at P21, the structure of filiform papillae was identical to that in the adult. These results indicate that, in rats, the morphogenesis of filiform papillae advances in parallel with keratinisation of the lingual epithelium from just before birth to a few weeks after birth. PMID- 10473291 TI - Temporal expression of the matrix metalloproteinase MMP-2 correlates with fibronectin immunoreactivity during the development of the vascular system in the chick embryo chorioallantoic membrane. AB - In this study we have examined in the chick embryo chorioallantoic membrane (ChAM) the expression of the matrix metalloproteinase 2 (MMP-2) and have correlated this parameter with the expansion of the ChAM vasculature and with the expression of 3 extracellular matrix components (fibronectin, laminin and type IV collagen), which differentially modulate angiogenesis. In the early phases of ChAM development, between d 6 and d 8 of incubation, when the increase of the ChAM vasculature is maximal, higher values of MMP-2 and, respectively, of fibronectin immunoreactive area, are detectable. These results indicate that MMP 2 activity and fibronectin expression are 2 strictly related components of angiogenesis occurring in vivo. PMID- 10473292 TI - The surface contour of articular cartilage in an intact, loaded joint. AB - The friction coefficients measured in diarthrodial joints are small. Theories of joint lubrication attribute this efficiency to entrapment or movement of synovial fluid, yet anatomical models of the surface are based on studies of isolated fragments of cartilage, not functional joints. To investigate the functional interrelationship of joint surfaces and synovial fluid, the ultrastructure of loaded joints was examined. Twenty-four New Zealand white rabbit knee joints were loaded either statically or moved ex vivo using simulated muscle forces and then plunge-frozen under load. After fixation in the frozen/loaded state by freeze substitution fixation, the medial joint compartments were embedded in epoxy resin while still articulated. Bone was trimmed away from the articular surfaces, permitting the cartilage to be sectioned for light and electron microscopy. These joint surfaces were then compared with controls which were not loaded, not moved or had been disarticulated prior to embedding. Articular surfaces of loaded joints were smooth at magnifications from x 35 to x 7500, whereas the tibial surfaces of nonloaded joints were irregular. Small pools of joint fluid were observed at the meniscal edge and beneath the anterior horn of the meniscus. At magnifications of x 40000, the joint surfaces were separated by a uniform 100 nm space containing fluid. An amorphous, electron dense articular surface lamina was present but, when loaded, was thicker and flatter than previously reported. No surface pits or bumps were visible in embedded, loaded joints. This is the first ultrastructural study of intact loaded joints. The findings suggest that fluid film lubrication is present in diarthrodial joints, but the fluid sequestration postulated in several models is not apparent. PMID- 10473293 TI - Median artery revisited. AB - This study confirms that the median artery may persist in adult life in 2 different patterns, palmar and antebrachial, based on their vascular territory. The palmar type, which represents the embryonic pattern, is large, long and reaches the palm. The antebrachial type,which represents a partial regression of the embryonic artery is slender, short, and terminates before reaching the wrist. These 2 arterial patterns appear with a different incidence. The palmar pattern was studied in the whole sample (120 cadavers) and had an incidence of 20%, being more frequent in females than in males (1.3:1), occurring unilaterally more often than bilaterally (4:1) and slightly more frequently on the right than on the left (1.1:1). The antebrachial pattern was studied in only 79 cadavers and had an incidence of 76%, being more frequent in females than in males (1.6:1); it was commoner unilaterally than bilaterally (1.5:1) and was again slightly more prevalent on the right than on the left (1.2:1). The origin of the median artery was variable in both patterns. The palmar type most frequently arose from the caudal angle between the ulnar artery and its common interosseous trunk (59%). The antebrachial pattern most frequently originated from the anterior interosseous artery (55%). Other origins, for both patterns, were from the ulnar artery or from the common interosseous trunk. The median artery in the antebrachial pattern terminated in the upper third (74%) or in the distal third of the forearm (26%). However, the palmar pattern ended as the 1st, 2nd or 1st and 2nd common digital arteries (65%) or joined the superficial palmar arch (35%). The median artery passed either anterior (67%) or posterior (25%) to the anterior interosseous nerve. It pierced the median nerve in the upper third of the forearm in 41% of cases with the palmar pattern and in none of the antebrachial cases. In 1 case the artery pierced both the anterior interosseous and median nerves. PMID- 10473294 TI - The porcine bronchial artery. Anastomoses with oesophageal, coronary and intercostal arteries. AB - Information about the existence and anatomy of arterial anastomoses with the porcine bronchial artery is lacking in the literature. Prior to basic physiological investigations in a porcine model related to lung transplantation with bronchial artery revascularisation, this study was designed to examine the anatomy of systemic arterial anastomoses with the bronchial artery system. Twenty pigs were studied in 3 groups. In 2 groups the heart-lung block was removed with all mediastinal structures. One group served for investigation of coronary bronchial artery anastomoses and one for investigation of oesophageal-bronchial artery anastomoses. The systemic arteries to be examined were cannulated. The inflated heart-lung block was examined macroscopically with Evans blue, and radiographically after contrast injection. In the 3rd group intercostobronchial artery anastomoses were studied radiographically with the heart-lung block in situ. Coronary-bronchial artery anastomoses were demonstrated in 3 of the 5 pigs with an aortic 'pouch' technique, but contrast was very limited in 2 of these 3. Oesophageal arterial anastomoses with bronchial arterial branches and/or the pulmonary veins were demonstrated in 6 of the 7 pigs and more markedly than the coronary-bronchial anastomoses. Intercostobronchial artery anastomoses could not be demonstrated angiographically. It was concluded that the existence of coronary bronchial and oesophageal-bronchial artery anastomoses in the pig appear to establish an arterial net between the base of the heart and the distal oesophagus. The resemblance to human oesophageal-bronchial artery anastomoses supports use of a porcine model for experimental studies. PMID- 10473295 TI - Angiotensin II is a growth factor in the peri-implantation rat embryo. AB - Angiotensin II (ANG II) is increasingly recognised as a growth factor, both in its own right and through interactions with other growth factors. There is a high density of ANG II receptors in the rat fetus, especially the AT2 receptor, the function of which is still uncertain. We have now studied the effects of ANG II on growth and development in the rat embryo in vitro between d 9.5 and 11.5, and characterised the receptor subtype mediating these effects. Embryos were cultured in whole rat serum, a high molecular weight retenate after ultrafiltration of whole rat serum, retenate with angiotensin II and retenate with ANG II and AT1 or AT2 receptor blockers. Growth and development were scored using conventional methods. Culture in retenate was associated with a marked reduction in growth and development by comparison with whole rat serum. This was partly, and significantly (P < 0.001), reversed by angiotensin II. The optimum concentration of angiotensin II was found to be angiotensin II 10(-11) M, within the physiological range. Angiotensin II had highly significant effects on both somatic (P < 0.001) and yolk sac/allantoic (P < 0.005) development. The latter effects suggest a role for angiotensin II in placentation. The effects of angiotensin II were blocked by PD123319, an AT2 blocker, but not by GR117289, an AT1 blocker. Interestingly, culture in retenate with GR117289 without added angiotensin II was also associated with some increase in growth (P < 0.05). Angiotensin II in low concentrations was measurable in the retenate, presumably arising from the action of endogenous renin on angiotensinogen. We therefore postulate that this effect of GR117289 was due to the action of endogenous angiotensin II on 'uncovered' AT2 receptors. This study has thus demonstrated a direct growth promoting effect of angiotensin II during organogenesis in the whole rat embryo in vitro. This effect is mediated through the AT2 receptors. PMID- 10473296 TI - Distribution patterns of vimentin-immunoreactive structures in the human prosencephalon during the second half of gestation. AB - Neuronal migration is guided by long radially oriented glial fibres. During late stages of development radial glial cells are transformed into astrocytes. A predominant intermediate filament protein within radial glial cells and immature astrocytes is vimentin. In this study fetal brain sections were used to demonstrate the transient features of vimentin-positive radial glia. In the lower half of the cerebral wall of the 6th gestational month bundles, curvature, and crossing of vimentin-positive fibres are regularly seen. Moreover, fibres terminating on vessels are observed. In the upper half fibres are radially oriented; when ascending towards the pial surface the number and diameter of fibres appears conspicuously decreased. Radially aligned fibres display numerous varicosities. In the 8th month the bulk of vimentin-positive fibres is encountered next to the ganglionic eminence and below isocortical cerebral fissures. The dentate gyrus is conspicuous due to its high amount of immunolabelled fibres. Furthermore, densely packed fibres are visible within the internal and external capsule and in the vicinity of the anterior commissure. Radial glial somata are found in the proliferative areas as well as in the adjacent white matter. In the latter location bipolar, monopolar and stellate vimentin-positive cells are present. The results demonstrate an area-specific distribution pattern of vimentin-positive structures which can be correlated with migrational events. Areas maturing late in development for instance, reveal dense immunolabelling in the 8th month. The orientation and position of radial fibres point to an additional developmental role of these fibres, i.e. their involvement in the guidance of growing axons. Moreover, the arrangement and morphology of vimentin-positive fibres, such as retraction of fibres or occurrence of varicosities, are indicative of degenerative events. Accordingly, a transformation of radial glial somata, their displacement towards the white matter and finally the growth of stellate processes can clearly be demonstrated. PMID- 10473297 TI - The electron microscope appearance of the subchondral bone plate in the human femoral head in osteoarthritis and osteoporosis. AB - The subchondral bone plate supports the articular cartilage in diarthrodial joints. It has a significant mechanical function in transmitting loads from the cartilage into the underlying cancellous bone and has been implicated in the destruction of cartilage in osteoarthritis (OA) and its sparing in osteoporosis (OP), but little is known of its composition, structure or material properties. This study investigated the microscopic appearance and mineral composition of the subchondral bone plate in femoral heads from patients with OA or OP to determine how these correspond to changes in composition and stiffness found in other studies. Freeze-fractured full-depth samples of the subchondral bone plate from the femoral heads of patients with osteoarthritis, osteoporosis or a matched control group were examined using back scattered and secondary emission scanning electron microscopy. Other samples were embedded and polished and examined using back-scattered electron microscopy and electron probe microanalysis. The appearances of the samples from the normal and osteoporotic patients were very similar, with the subchondral bone plate overlayed by a layer of calcified cartilage. Osteoporotic samples presented a more uniform fracture surface and the relative thicknesses of the layers appeared to be different. In contrast, the OA bone plate appeared to be porous and have a much more textured surface. There were occasional sites of microtrabecular bone formation between the trabeculae of the underlying cancellous bone, which were not seen in the other groups, and more numerous osteoclast resorption pits. The calcified cartilage layer was almost absent and the bone plate was apparently thickened. The appearance of the osteoarthritic subchondral bone plate was, therefore, considerably different from both the normal and the osteoporotic, strongly indicative of abnormal cellular activity. PMID- 10473298 TI - Ultrastructure of intramural ganglia in the striated muscle portions of the guinea pig oesophagus. AB - The ultrastructure of the myenteric plexus located in the striated muscle portion of the guinea pig oesophagus was examined and compared with that of the plexus associated with the smooth muscle portion of the rest of the digestive tract. The oesophageal ganglia had essentially the same architecture as those of the smooth muscle portion, such as a compact neuropil without the intervention of connective tissue and blood vessels. Some features, however, were particular to the striated muscle part of the oesophagus. It was clearly demonstrated that myelinated fibres, probably sensory terminals of vagal origin, join the myenteric ganglia. Synapses and terminal varicosities are sparsely distributed within the ganglia and fewer morphological types of axon varicosities could be distinguished compared with other regions. Glial cells are well developed in the oesophageal myenteric ganglia. These cells outnumber the ganglion cells, having a higher ratio than in the lower digestive tract, and form numerous cytoplasmic lamellar processes. The lamellar processes, located at the surface of the ganglia, considerably reduce the area of neuronal membrane which directly contacts the basal lamina. The role of these lamellar processes in the oesophageal ganglia is discussed. PMID- 10473299 TI - Electron-immunocytochemical studies of perivascular nerves of mesenteric and renal arteries of golden hamsters during and after arousal from hibernation. AB - Electron immunocytochemistry was used to examine perivascular nerves of hamster mesenteric and renal arteries during hibernation and 2 h after arousal from hibernation. Vessels from cold-exposed but nonhibernating, and normothermic control hamsters were also examined. During hibernation the percentage of axon profiles in mesenteric and renal arteries that were immunopositive for markers of sympathetic nerves, tyrosine hydroxylase (TH) and neuropeptide Y (NPY), were increased 2-3 fold compared with normothermic and cold control animals. This increase was reduced markedly only 2 h after arousal from hibernation. The small percentage of nitric oxide synthase-1-positive axon profiles found in mesenteric (but not renal) arteries was also increased during hibernation and returned towards control values after arousal. In contrast, the percentage of perivascular axons immunostaining for vasoactive intestinal polypeptide (VIP), a marker for parasympathetic nerves, was reduced in mesenteric arteries during hibernation. There was no labelling of perivascular nerves for substance P in either mesenteric or renal arteries. It is suggested that the increase in percentage of TH- and NPY-immunostained perivascular nerves may account for the increased vasoconstriction associated with high vascular resistance that is known to occur during hibernation. The reduction in the percentage of axons positive for VIP in hibernating animals would contribute to this mechanism since this neuropeptide is a vasodilator. PMID- 10473300 TI - Hippocampal layers on high resolution magnetic resonance images: real or imaginary? AB - To evaluate whether the lines occasionally detected on clinical magnetic resonance (MR) images are genuine hippocampal layers, a formalin fixed hippocampal specimen was scanned using T2 weighted sequences at 7 Tesla (voxel dimensions 0.064 x 0.064 x 1 mm) and at 1.5 Tesla (voxel dimensions: 0.156 x 0.156 x 1 mm) and compared with the results of histological examination. In addition, a healthy volunteer was scanned with a T2 weighted sequence at 1.5 Tesla (voxel dimensions: 0.469 x 0.469 x 2 mm). On 7 Tesla images hippocampal layers and the granule cell layer of the dentate were visible. On 1.5 Tesla images of the specimen, the hippocampal layers were again identified, but the granule cell layer of the dentate was not detectable. On 1.5 Tesla images of the hippocampus in vivo, 3 layers could be distinguished in the hippocampus on some slices. These mainly represented the alveus, pyramidal cell layer and stratum radiatum. A dark line consisting of a few pixels possibly represented the dentate gyrus. Our results show that the lines occasionally detected on clinical MR images are likely to be real hippocampal layers. However, the resolution currently used in clinical imaging (typically 0.469 x 0.469 x 2 mm or lower) is not sufficient for the detection of all hippocampal layers. For the reliable detection of all hippocampal layers on MR images an increase by a factor of approximately 20 would be necessary. PMID- 10473301 TI - Compartmentalisation of the developing trigeminal ganglion into maxillary and mandibular divisions does not depend on target contact. AB - During development axons contact their target tissues with phenomenal accuracy but the mechanisms that control this homing behaviour remain largely elusive. A prerequisite to the study of the factors involved in hard-wiring the nervous system during neurogenesis is an accurate calendar of developmental events. We have studied the maxillary and mandibular components of the trigeminal system to determine the stages during embryogenesis when a gross somatotopic order is first established within the trigeminal ganglion and the axons projecting to the brainstem. The retrograde transganglionic fluorescent tracers DiO and DiI were injected into the maxillary and mandibular arches or their derivatives in fixed mouse embryos staged between 13 and 40 somites (E9-E11). After 1-4 wk, the distribution of the 2 tracers was determined using confocal laser scanning microscopy. The first maxillary nerve cell bodies and their developing axons were labelled at the 30 somite stage (E10). This was 2 somite stages earlier than the mesencephalic nucleus and the ganglion cell bodies of the mandibular nerve. The gross somatotopic division of cells within the trigeminal ganglion projecting to the maxillary and mandibular targets was established by the 32 somite stage (E10). This arrangement was evident as 2 groups of cell bodies occupying adjacent but separate regions of the trigeminal ganglion. The central branches of the maxillary and mandibular cell bodies entered the metencephalon as 2 distinct bundles at the same stage. The trigeminal motor nucleus was first detected at the 38 somite stage (E10.5). Gross somatotopy in the major divisions of the trigeminal ganglion is established before outgrowing axons have contacted their peripheral target tissue at E10.5. This suggests that target tissues do not induce somatotopy. PMID- 10473302 TI - Characteristic intraepidermal nerve fibre endings of the intervibrissal fur in the mystacial pad of the rat: morphological details revealed by intravital methylene blue staining and the zinc iodide-osmium tetroxide technique. AB - Light microscopic observations employing intravital methylene blue staining and impregnation by the zinc iodide-osmium tetroxide technique are presented for intraepidermal nerve fibre endings of the intervibrissal fur in the mystacial pad of the rat snout. Both procedures revealed anatomical details of the intraepidermal nerve fibre plexus in epidermal hillocks often located very close to the mouths of hairs. These nerve fibres appeared to resemble those described in previous immunohistochemical studies as cluster or bush endings. The methylene blue preparations demonstrated the existence of an intensely stained enlargement at the site of the branching point of the nerve fibres which seemed to be functionally related to the development of such nerve fibre plexuses. Due to their close association with hairs, these nerve fibre plexuses are most likely to be mechanoreceptive. Additionally, solitary varicose nerve fibres were found loosely distributed within the epidermis. The visualisation of 2 different morphological types of nerve fibre endings extends the validity of the concept of punctate sensibility into the epidermis. Methylene blue staining appeared to be somewhat superior to the zinc iodide-osmium tetroxide technique. Due to their selectivity for intraepidermal nerve fibres, the methods applied here supplement immunohistochemical procedures in a helpful manner. PMID- 10473303 TI - Translating ethical principles into outcome measures for mental health service research. AB - BACKGROUND: Mental health service research continues to use only outcome measures that are available rather than develop measures that are important. This paper argues that it is necessary to select and then define a set of ethical principles that can be operationalized and validated as outcome measures to provide a wider balance of information for health policy and clinical service decisions. METHODS: The method used is to adopt a five stage procedure: (i) to select ethical principles most directly relevant for mental health services and their evaluation at the local level; (ii) to propose definitions of these principles; (iii) to validate these definitions; (iv) to translate the defined principles into operationalized outcome measures; and (v) to use these outcome measures in mental health services research, within the context of evidence-based medicine. RESULTS: We address steps (i) and (ii) of this five-stage procedure. Nine principles are selected and defined: autonomy, continuity, effectiveness, accessibility, comprehensiveness, equity, accountability, coordination and efficiency. These principles can together be referred to as the three ACEs. CONCLUSIONS: Of these nine principles, only two (effectiveness and efficiency) have so far been fully translated into quantitative outcome measures, upon which the evidence-based medicine approach depends. We propose that further concepts also be developed into a more complete multidimensional range of fully operationalized outcome measures. PMID- 10473304 TI - The epidemiology of autism: a review. AB - BACKGROUND: There is some uncertainty about the rate and correlates of autism. METHOD: Twenty-three epidemiological surveys of autism published in the English language between 1966 and 1998 were reviewed. RESULTS: Over 4 million subjects were surveyed; 1533 subjects with autism were identified. The methodological characteristics of each study are summarized, including case definition, case finding procedures, participation rates and precision achieved. Across surveys, the median prevalence estimate was 5.2/10000. Half the surveys had 95% confidence intervals consistent with population estimates of 5.4-5.5/10000. Prevalence rates significantly increased with publication year, reflecting changes in case definition and improved recognition; the median rate was 7.2/10 000 for 11 surveys conducted since 1989. The average male/female ratio was 3.8:1, varying according to the absence or presence of mental retardation. Intellectual functioning within the normal range was reported in about 20% of subjects. On average, medical conditions of potential causal significance were found in 6% of subjects with autism, with tuberous sclerosis having a consistently strong association with autism. Social class and immigrant status did not appear to be associated with autism. There was no evidence for a secular increase in the incidence of autism. In eight surveys, rates for other forms of pervasive developmental disorders were two to three times higher than the rate for autism. CONCLUSION: Based on recent surveys, a minimum estimate of 18.7/10000 for all forms of pervasive developmental disorders was derived, which outlines the needs in special services for a large group of children. PMID- 10473305 TI - Recall of depressive episode 25 years previously. AB - BACKGROUND: Lifetime rates of depression reported in epidemiological surveys are generally only twice the 12 month rates. Either people forget the symptoms of depression or many people who have a depressive episode remain depressed for many years. Both may be true. There is a need to examine the long-term clinical validity of interviews that are used to make lifetime diagnoses. METHODS: Forty five patients who were part of a long-term follow-up study of depression were interviewed 25 years after the index episode. The diagnoses from the original, fully structured interviews were compared with the responses people made for that period when interviewed using the CIDI 25 years later. RESULTS: Twenty-seven patients met CIDI DSM-III-R criteria for depression at index episode. At the 25 year follow-up, 19 of the 27 reported the essential symptoms of 'depression or loss of interest' being present at the index time, and in 14 of the 27 the depressive symptoms recalled met criteria for DSM-III-R major depressive episode at that time. CONCLUSIONS: Seventy per cent of people who were hospitalized for a major depressive episode can recall being depressed but only half can recall sufficient detail to satisfy the diagnostic criteria when interviewed 25 years later. As depressive episodes, especially those severe enough to warrant admission, are recalled better than many other diagnoses, one must be cautious about the lifetime rates for mental disorders reported in retrospective epidemiological surveys. PMID- 10473306 TI - A randomized controlled trial of individual psychological debriefing for victims of violent crime. AB - BACKGROUND: It has been suggested that giving people the opportunity talk about a traumatic experience may prevent the development of later disorder. We tested the efficacy of two brief interventions, education and psychological debriefing, designed to prevent adverse psychological reactions to criminal victimization. METHODS: Individuals who had been the victims of a violent crime within the past month were written to and invited to take part in a study of their attitudes to crime and punishment: 2161 were contacted and 243 replied, of whom 157 were eligible and were randomly assigned either to an education condition, to a psychological debriefing plus education condition, or to an assessment only condition. Education involved providing information about normal post-traumatic reactions. Debriefing involved in-depth probing about events, thoughts and feelings experienced during the crime. Subjects were recruited from police and hospital sources and interviewed in their own homes: 138 were followed up at 6 months, and 92 at 11 months. RESULTS: Outcome was assessed using a DSM-III-R diagnosis of PTSD, the Post-traumatic Symptom Scale, the Impact of Event Scale and the Beck Depression Inventory. All groups improved over time but there were no between-group differences. CONCLUSIONS: No evidence was found to support the efficacy of brief one-session interventions for preventing post-traumatic symptoms in individual victims of violent crime. PMID- 10473307 TI - Relatives' expressed emotion (EE) and PTSD treatment outcome. AB - BACKGROUND: Expressed emotion (EE) is a measure that has been used to assess the quality of the relationship between patient and their key relative. It has been shown to be strongly predictive of clinical outcome in a range of psychiatric and medical disorders. This study investigated the effect of EE on treatment outcome in chronic post-traumatic stress disorder (PTSD). METHODS: A prospective design was adopted. The key relatives of 31 PTSD patients participating in a treatment trial comparing imaginal exposure with cognitive therapy were interviewed and rated on EE prior to treatment allocation. The effect of EE on post-treatment clinical outcomes was assessed. RESULTS: Sixteen patients (52%) had high EE and 15 (48%) low EE relatives. Patients with high EE relatives showed lesser change scores on the main outcome variable of the trial, the total CAPS score, and on all the secondary outcome variables than those with low EE relatives. Using different multiple regression models the EE scales of criticism and hostility predicted just under 20% of the outcome variance. These two scales were highly correlated and criticism marginally predicted the greatest variance (19.7%). CONCLUSIONS: The results highlight the importance of the quality of the patient's social environment in influencing their response to cognitive and behavioural treatments. PMID- 10473309 TI - Evaluating psychiatric morbidity in a general hospital: a two-phase epidemiological survey. AB - BACKGROUND: This study aimed to assess psychiatric morbidity and to collect information on disability, life events and family support in a representative sample of patients admitted to a general hospital. METHODS: On the basis of information collected in a pilot study a systematic sample of patients consecutively admitted to seven general medical and seven surgical wards of the Academic General Hospital of Verona was selected and interviewed using a two phase screening procedure and standardized instruments (GHQ-12, HADS, BDQ and CIDI-PHC). All data were analysed using appropriately weighted logistic regression procedures. RESULTS: A total of 1039 patients completed the GHQ-12 and 298 (28.7%) were high-scorers: 363 patients were interviewed with CIDI-PHC. The prevalence of ICD-10 cases was 26.1%. The most common psychiatric diagnoses were current depression (12.8%) and generalized anxiety disorder (10.8%), followed by alcohol related disorders (5 %). A higher prevalence of ICD-10 cases was found in medical wards, among females, patients older than 24 years, unemployed and separated/divorced people. Life events were associated with psychopathology, and so was the number of disability days. Although 49.8% of ICD-10 cases were identified by the hospital doctors as having a psychological disorder, 23.1% of ICD-10 cases were referred to the liaison psychiatric service. CONCLUSION: The results of the present study stress the need to collect epidemiologically-based data on psychological disorders and their recognition not only in general practice, but also in general hospital settings, in order to have a more complete picture of the pathways to specialist care. PMID- 10473308 TI - Vulnerability to assaultive violence: further specification of the sex difference in post-traumatic stress disorder. AB - BACKGROUND: We examine potential sources of the sex differences in post-traumatic stress disorder (PTSD) in the community. METHODS: Data were obtained from a representative sample of 2181 persons aged 18-45 years in the Detroit primary metropolitan statistical area, which is a six-county area containing more than four million residents. A random digit dialling method was used to select the sample and a computer-assisted telephone interview was used to obtain the data. DSM-IV PTSD was assessed with respect to a randomly selected trauma from the list of qualifying traumas reported by each respondent. RESULTS: The lifetime prevalence of exposure and the mean number of traumas were lower in females than males. The overall conditional risk of PTSD (i.e. the probability of PTSD among those exposed to a trauma) was approximately twofold higher in females than males, adjusting for the sex difference in the distribution of trauma types. The sex difference was due primarily to females' greater risk following assaultive violence. The sex difference in the avoidance and numbing symptom group following assaultive violence exceeded the sex differences in other symptom groups. CONCLUSIONS: Future research should focus on sex differences in the response to assaultive violence, including potential explanations for females' greater probability to experience avoidance and numbing. PMID- 10473310 TI - Training primary-care physicians to recognize, diagnose and manage depression: does it improve patient outcomes? AB - BACKGROUND: We developed a comprehensive, 20-hour training programme for primary care physicians, that sought to improve their ability to detect, diagnose and manage depression. We evaluated the effects of physician training on patient outcomes, using a pre-post design. METHODS: In the pre-training phase of the study, we sampled 1834 consecutive patients of 17 primary-care physicians and evaluated 518 of these patients for the presence of depression. We measured outcomes of all patients with depression at 3 months and 1 year. The outcome measures were: severity of psychopathology; duration of depressive episode; and level of daily functioning. After the 17 physicians completed the training, we drew a new sample from their practices (498 of 1785 consecutive patients were evaluated for depression) and measured outcomes for the depressed patients. RESULTS: We found an effect of the training on short-term outcome, particularly for patients with a recent-onset depression. At 3-month follow-up depressed patients whose physicians had received training had less severe psychopathology and patients with recent-onset depression also showed higher levels of daily functioning than patients of the same physicians prior to the training. The patients with a recent-onset depression that was recognized by trained physicians had shorter depressive episodes, but this was not statistically significant. At 1 year follow-up, all training effects had faded away. CONCLUSIONS: Training primary-care physicians to recognize, diagnose and manage depression can improve short-term patient outcomes, especially for patients with a recent onset of depression. Patients suffering from a recurrent or chronic depression may need more specific interventions, both for acute treatment and long-term management. PMID- 10473312 TI - The temporal stability and co-morbidity of prolonged fatigue: a longitudinal study in primary care. AB - BACKGROUND: Depression, anxiety and fatigue are among the most common symptoms presented in primary care. Whether such symptoms indicate discrete psychological syndromes or whether they result from a common vulnerability is not clear. This study examined longitudinally the patterns of co-morbidity between prolonged fatigue and other forms of psychological distress in patients attending general practitioners. METHODS: Adults attending primary care completed questionnaires designed to detect cases of prolonged fatigue and psychological distress at presentation and 12 months later. RESULTS: Of 652 patients, the prevalence rates of 'prolonged fatigue' alone, 'psychological distress' alone, 'prolonged fatigue + psychological distress' and 'no disorder' were 7%, 19%, 15% and 59% respectively at initial assessment. Of those patients with any prolonged fatigue syndrome initially, 58% still reported fatigue 12 months later (representing 13% of the total sample). Most importantly, the risk of developing prolonged fatigue was not increased in patients who initially had psychological distress (OR = 0.95; 95% CI 0.2-3.6), neither was the risk of developing psychological distress increased in patients who initially had prolonged fatigue (OR = 1.4; 95% CI 0.6 3.4). CONCLUSIONS: This study indicates that prolonged fatigue is a persistent diagnosis over time. The longitudinal patterns of co-morbidity with psychological distress do not support an aetiological model that proposes a common vulnerability factor for these disorders. Psychiatric classification systems may be better served by treating prolonged fatigue and psychological distress as independent disorders. PMID- 10473311 TI - Onset of disability in depressed and non-depressed primary care patients. AB - BACKGROUND: While cross-sectional and longitudinal studies have consistently found depressive illness and disability to be related, understanding whether depression leads to subsequent onset of disability is limited. METHODS: In the context of the multi-centre international WHO Collaborative Study on Psychological Problems in General Health Care, we followed prospectively consulting non-elderly primary care patients who were essentially disability free at baseline but who differed in baseline depression status, comprising 1051 patients free of physical disability at baseline including 14% depression; 914 free of social disability including 9% depression. Depression status was assessed with the CIDI; patient-reported physical disability with the MOS physical functioning scale and social disability with the BDQ role functioning and number of disability days measures; investigator-rated social disability with the Occupational section of Groningen Social Disability schedule; and the treating physicians rated the severity of physical illness. RESULTS: In patients essentially disability free at baseline, depressive illness resulted in a 1.5 fold (at 3 months) and a 1.8-fold (at 12 months) increase in risk of onset of physical disability, after controlling for physical disease severity. Depressive illness also resulted in a 2.2-fold (at 3 months) and a 23-fold (at 12 months) increase in risk of onset of social disability, after controlling for physical disease severity, physical disability and onset of physical disability. CONCLUSIONS: Among non-elderly primary care patients, depressive illness is associated with onset of physical disability and shows an even stronger association with onset of social disability. PMID- 10473313 TI - Fatigue and psychiatric disorder: different or the same? AB - BACKGROUND: Fatigue and psychiatric symptoms are common in the community, but their association and outcome are sparsely studied. METHOD: A total of 1177 patients were recruited from UK primary care on attending their general practitioner. Fatigue and psychiatric disorder was measured at three time points with the 12-item General Health Questionnaire and the 11-item Fatigue Questionnaire. RESULTS: Total scores for fatigue and psychiatric disorder did not differ between the three time points and were closely correlated (r around 0.6). The association between non-co-morbid ('pure') fatigue and developing psychiatric disorder 6 months later was the same as that for being well and subsequent psychiatric disorder. Similarly, having non-co-morbid psychiatric disorder did not predict having fatigue any more than being well 6 months previously. Between 13 and 15% suffered from non-co-morbid fatigue at each time point and 2.5% suffered from fatigue at two time points 6 months apart. Less than 1% of patients suffered from non-co-morbid fatigue at all three time points. CONCLUSIONS: The data are consistent with the existence of 'pure' independent fatigue state. However, this state is unstable and the majority (about three-quarters) of patients become well or a case of psychiatric disorder over 6 months. A persistent, independent fatigue state lasting for 6 months can be identified in the primary-care setting, but it is uncommon of the order of 2.5%. Non-co-morbid (pure) fatigue did not predict subsequent psychiatric disorder. PMID- 10473314 TI - The Range of Impaired Functioning Tool (LIFE-RIFT): a brief measure of functional impairment. AB - BACKGROUND: The literature documents that functional impairment is associated with affective disorders. Nevertheless, the choice among thorough, yet brief, well-validated assessments of functional impairment is limited. The objective of this study was to evaluate the psychometric properties of a brief scale of functional impairment, the Range of Impaired Functioning Tool (LIFE-RIFT). METHOD: The study sample included subjects who presented with major depressive disorder at intake into the NIMH Collaborative Depression Study (CDS). The LIFE RIFT is composed of items that are included in the Longitudinal Interval Follow up Evaluation (LIFE). The reliability and validity were examined using data from LIFE-RIFT assessments conducted at four points in time: 6, 12, 18 and 24 months after intake into the CDS. RESULTS: Cross-sectional one factor models accounted for the covariance structure among the four scale items. A longitudinal factor model, with an invariant factor structure over time, also fitted the data well and indicated that the scale items are measures of one construct, namely functional impairment. The internal consistency reliability of the scale was supported with alpha coefficients ranging from 0.81 to 0.83. The inter-rater reliability intraclass correlation coefficient (ICC) was 0.94. Mixed-effect linear regression models showed that those in episode were significantly more impaired than those in recovery. Furthermore, in analyses of predictive validity, impairment was positively associated with subsequent recurrence and negatively associated with subsequent recovery. CONCLUSIONS: This psychometric evaluation provides empirical support for the reliability and validity of the LIFE-RIFT, a brief measure of functional impairment. PMID- 10473315 TI - Scales to measure dimensions of hallucinations and delusions: the psychotic symptom rating scales (PSYRATS). AB - BACKGROUND: Scales to measure the severity of different dimensions of auditory hallucinations and delusions are few. Biochemical and psychological treatments target dimensions of symptoms and valid and reliable measures are necessary to measure these. METHOD: The inter-rater reliability and validity of the Psychotic Symptom Rating Scales (PSYRATS: auditory hallucination subscale and delusions subscale), which measure several dimensions of auditory hallucinations and delusions were examined in this study. RESULTS: The two scales were found to have excellent inter-rater reliability. Their validity as compared with the KGV scale (Krawiecka et al. 1977) was explored. CONCLUSIONS: It is concluded that the PSYRATS are useful assessment instruments and can complement existing measures. PMID- 10473316 TI - Reliability of the Salford Needs Assessment Schedule for Adolescents. AB - BACKGROUND: For adolescents, there is no specific needs assessment instrument that assesses significant problems that can benefit from specified interventions. A new instrument (S.NASA) was developed by incorporating and adapting three well established adult needs assessment instruments. The S.NASA covers 21 areas of functioning including social, psychiatric, educational and life skills. METHOD: Client and carer interviews were conducted by different researchers. A week later the interviews were repeated using a crossover design. Significant (cardinal) problems were generated from the clinical interviews using a pre-defined algorithm. Final need status (three categories) was made by clinicians assessing the cardinal problems against defined interventions. The interventions were generated from discussions with clinicians and a survey of appropriate professionals working with adolescents. RESULTS. Pre-piloting led to the final version being administered to 40 adolescents from secure units, forensic psychiatric and adolescent psychiatric services. There were 25 males and 15 females, mean age 15.5 years. Overall there were moderate to good inter-rater and test-retest reliability coefficients, the test-retest reliability coefficients for the total scores on the needs assessment interviews ranged from 0.73 to 0.85. Consensual and face validity was good, the adolescents and staff finding the instrument useful and helpful. CONCLUSIONS: This new needs assessment instrument shows acceptable psychometric properties. It should be of use in research projects assessing the needs and the provision of services for adolescents with complex and chronic problems. PMID- 10473317 TI - Source monitoring deficits in patients with schizophrenia; a multinomial modelling analysis. AB - BACKGROUND: Schizophrenia patients, particularly those with symptoms such as thought insertion, passivity experiences and hallucinations, may share an underlying cognitive deficit in monitoring the generation of their own thoughts. This deficit, which has been referred to as 'autonoetic agnosia', may result in the conclusion that self-generated thoughts come from an external source. Previous work supports this notion, yet the statistical approaches that have been used have not enabled a distinction between specific deficits suggesting autonoetic agnosia and more general cognitive dysfunction. METHODS: Autonoetic agnosia was assessed using source-monitoring paradigms in 28 patients with schizophrenia and 19 control subjects. Multinomial model analyses, which allow the distinction between deficits in recognizing information, remembering its source, and response biases, were applied to the data. RESULTS: Schizophrenia patients were impaired in discriminating between words that came from two external sources, from two internal sources, and one internal and one external source. In a condition requiring subjects to distinguish between words they had heard from those they had imagined hearing, when schizophrenic patients did not remember the source of the information, they showed a stronger bias than controls to report that it had come from an external source. CONCLUSIONS: The application of multinomial models to source monitoring data suggests that schizophrenia patients have source monitoring deficits that are not limited to the distinction between internally-generated and externally-perceived information. However, when schizophrenia patients do not remember the source of information, they may be more likely than controls to report that it came from an external source. PMID- 10473318 TI - Duration of illness and structure of symptoms in schizophrenia. AB - BACKGROUND: Previous research has mainly focused on the cross-sectional structure of symptoms in schizophrenia. This meta-analysis examined the association of duration of illness with the structure of symptoms. METHODS: Using explicit criteria, 22 studies reporting on the correlations of symptoms in 2665 schizophrenic patients were selected. From each study, symptom-pair correlations for negative symptoms as rated by Scale for the Assessment of Negative Symptoms (SANS) and positive symptoms as rated by the Scale for the Assessment of Positive Symptoms (SAPS) were extracted. Variability among symptom-pair correlations across studies was assessed using tests of homogeneity. For symptom-pair correlations which were not found to be homogeneous, the association of average duration of illness with the symptom-pair correlations were examined. RESULTS: There was considerable variability in symptom-pair correlations across studies. Part of this variability was explainable by variations in average duration of illness. Longer duration of illness was associated with lower negative-negative symptom-pair correlations and higher negative-positive symptom-pair correlations. CONCLUSIONS: The findings suggest that the structure of symptoms in schizophrenia evolves over time, following a consistent pattern. In the early stages of illness, negative and positive symptoms form cohesive dimensions. With time, these dimensions become less cohesive and the boundaries between them, less clear. PMID- 10473319 TI - The structure of genetic and environmental risk factors for three measures of disordered eating. AB - BACKGROUND: The study explored the genetic and environmental risk factors for both the behaviours and attitudes characteristic of disordered eating. METHODS: In three waves of data collection, information was collected from female twins regarding their eating and attitudes towards eating, weight and shape. The first assessment consisted of a self-report questionnaire (1988-9) with 1682 women. The second assessment consisted of a semi-structured psychiatric interview schedule (1992-3), completed by 1852 women, many of whom had completed Wave 1 assessment. The third assessment, with 325 women chosen from Waves 1 and 2 (1995-6), consisted of a semi-structured interview (the Eating Disorder Examination). RESULTS: As only one twin pair was concordant for lifetime bulimia nervosa at Wave 3 assessment, ordinal measures of all assessments were used in a multivariate genetic analysis. Results indicated that additive genetic and non shared environmental influences best explained variance in liability to disordered eating, with about 60% (95% CI 50-68) of the variance explained by genetic factors. Comparison with a model allowing for the effects of shared environment indicated genetic factors accounted for a similar degree of variance (59%, 95% CI 36-68). CONCLUSION: Liability to the development of the behaviours and attitudes characteristic of eating disorders is best explained by both environmental and genetic factors, with covariation between the three measures best explained by a single latent phenotype of disordered eating which has a heritability of 60%. PMID- 10473320 TI - History of childhood sexual or physical abuse in Japanese patients with eating disorders: relationship with dissociation and impulsive behaviours. AB - BACKGROUND: Empirical data regarding prevalence of sexual and physical abuse histories in Japanese patients with eating disorders is lacking, in contrast to Western countries. This study investigated the prevalence of traumatic events in Japanese patients with eating disorders, and examined the relationship between such traumatic events and clinical features. METHODS: Subjects consisted of 33 patients with anorexia nervosa restricting type (RAN), 40 patients with anorexia nervosa binge eating/purging type (AN-BP), 63 patients with bulimia nervosa purging type (BN) and 99 healthy controls. All were female and diagnoses were based on DSM-IV. The Physical and Sexual Abuse Questionnaire (PSA), Eating Disorder Inventory (EDI) and Dissociation Experience Scale (DES) were administered to all of the subjects. RESULTS: Paradoxically, victims of minor sexual abuse committed by Chikan (a Japanese word indicating a person who commits minor sexual crimes) were more prevalent among controls than among patients with RAN, AN-BP or BN. However, physical punishment histories tended to be more prevalent among patients with AN-BP or BN than among RAN or controls. Only AN-BP and BN patients with physical punishment histories had twofold higher scores for DES and significantly more frequent histories of self-mutilation (67% v. 33%) compared with patients without such histories. CONCLUSION: An abuse history is not essential or a prerequisite to developing an eating disorder in Japan. PMID- 10473321 TI - The relationship among three models of personality psychopathology: DSM-III-R personality disorder, TCI scores and DSQ defences. AB - BACKGROUND: Current systems of describing personality pathology have significant shortcomings. A polydiagnostic approach is used to study the relationship between psychological, psychoanalytical and psychopathological models of personality. METHODS: The subjects were 256 patients enrolled in treatment studies of major depression and bulimia nervosa. Subjects were assessed using the Temperament and Character Inventory (TCI), the Defense Style Questionnaire (DSQ) and the Structured Clinical Interview for DMS-III-R personality disorders (SCID-II). RESULTS: Subjects had high rates of DSM-III-R personality disorders with 52% having at least one personality disorder. Cluster A personality disorders were correlated with low reward dependence, high harm avoidance and low self directedness and cooperativeness. Cluster B personality disorders were related to high novelty seeking and low self-directedness and cooperativeness. Cluster C personality disorders were correlated with high harm avoidance and low novelty seeking and low self-directedness. Immature defences were related to DSM-III-R personality symptoms, but individual defences were not related to personality clusters in a predictable way. Immature defences were strongly related to low self-directedness and cooperativeness. Both TCI self-directedness scores and immature defence scores were moderately predictive of the presence and number of personality disorders. CONCLUSION: A widely accepted clinical nosology (DSM-III-R personality disorders) rated using a clinical interview correlates reasonably predictably with two theoretical models derived from different paradigms and rated using self-reports. This might be seen as providing concurrent validity for all three models. However, serious methodological shortcomings confront studies of this type, including sample selection and measurement of personality dysfunction. One way to begin to resolve these problems is to study which personality measures are best related to treatment response and prognosis. PMID- 10473323 TI - Mid-sagittal anatomy in late-onset schizophrenia. AB - BACKGROUND: Of the midline brain structures, abnormalities have been demonstrated in the corpus callosum and cerebellum in young schizophrenic patients. Whether similar abnormalities are also present in late-onset schizophrenia (LOS) is not known. METHODS: The mid-sagittal cross-sectional areas of brain regions, in particular the corpus callosum and cerebellum, on magnetic resonance imaging were examined in a group of patients with late-onset schizophrenia (N = 25) and contrasted with two comparison groups - early-onset schizophrenia (EOS) (N = 24) and healthy volunteers (NC) (N = 30) matched for age and gender. RESULTS: While the mean corpus callosum area in the LOS group was smaller than in the EOS (by 10.2%) and NC (by 6.2%) groups, the three groups did not differ statistically in the corpus callosum area or the corpus callosum to cerebrum ratios. The cross sectional cerebellar areas or the cerebellum: cerebrum ratios also did not differ across the groups. The brainstem was smaller in the schizophrenic groups because of smaller cross-sectional areas of the pons, a statistically significant difference which could not be accounted for by any gross lesions on visual inspection. CONCLUSION: We found no abnormality in the mid-sagittal area of the corpus callosum and cerebellum in our early- or late-onset schizophrenia subjects. The significance of the finding of a smaller pontine cross-sectional area is unclear and speculation on it awaits independent replication using a volumetric measure. PMID- 10473322 TI - Criminality and antisocial behaviour in unselected men with sex chromosome abnormalities. AB - BACKGROUND: Previous studies on male patients with sex chromosome abnormalities (SCA), namely XYY and XXY, suggest that such patients commit criminal acts more frequently than expected. Most of these studies are affected by ascertainment bias. METHODS: Using a population-based sample of men with SCA, identified by screening 34380 infants at birth between 1967 and 1979, comparison between 16 XYY men, 13 XXY men and 45 controls were made in terms of frequency of antisocial personality disorder (APD) using the Schedule for Affective Disorders and Schizophrenia lifetime version. Rates of criminal convictions were examined in 17 XYY men, 17 XXY men and 60 controls. RESULTS: XYY males showed a significantly higher frequency of antisocial behaviour in adolescence and adulthood and of criminal convictions than the controls, but multiple regression analysis showed this to be mediated mainly through lowered intelligence. Property offences constituted the majority of offences in all groups. The XXY men did not show an increased rate of criminal convictions. It is possible that this apparently negative result relates to the relatively small numbers of cases and hence low power of this study. CONCLUSIONS: The findings of this study carry the advantage of not being affected by ascertainment bias and the disadvantage of having low power. It provides evidence for a slightly increased liability to antisocial behaviour in XYY men. PMID- 10473324 TI - Enhanced corticotropin response to corticotropin-releasing hormone as a predictor of mania in euthymic bipolar patients. AB - BACKGROUND: Dysregulation of corticotropin (ACTH) and cortisol response after corticotropin-releasing hormone (CRH) stimulation has been reported in bipolar patients. Most findings involve the pathophysiology of the depressive phase of the illness and its prediction. However, the possible predictive value of the CRH challenge test with respect to manic episodes remains unknown. METHODS: The ACTH and free cortisol response to the injection of 100 microg of synthetic human CRH and plasma cortisol-binding globulin levels were measured in 42 lithium-treated patients suffering from Research Diagnostic Criteria bipolar I disorder in remission, and 21 age- and sex-matched normal controls. A 1-year follow-up was conducted to assess any possible relationship between outcome and the hormonal response. RESULTS: Bipolar patients showed higher baseline and peak ACTH concentrations than control subjects. A higher area under ACTH concentration curve after CRH stimulation predicted manic/hypomanic relapse within 6 months by multiple regression analysis. CONCLUSION: Bipolar patients in remission show mild abnormalities in ACTH levels before and after CRH stimulation. CRH challenge may be a potentially good predictor of manic or hypomanic relapse in remitted bipolar patients. PMID- 10473325 TI - The relationship between depression and mortality in elderly subjects with less severe dementia. AB - BACKGROUND: Little is known about the effects of depression on the mortality rates of elderly subjects with dementia. METHODS: Logistic regression analysis, adjusting for possible confounders, was used to study the associations between GMS-AGECAT derived syndrome and symptom measures and 12-month mortality rates in a cohort of 73 elderly subjects who met the DSM-III-R criteria of dementia with a median MMSE score of 19. RESULTS: Twenty-three subjects (32%) died within the 12 month follow-up period. A baseline diagnosis of syndromal or subsyndromal depression was associated with increased mortality. At the symptom level mortality was predicted by higher scores on the factor 'mood symptoms'. The effects of interactions between depression measures and severity of dementia were not significant. CONCLUSIONS: Short-term mortality in elderly subjects with less severe dementia is predicted by the presence of (sub) syndromal depression and by mood symptoms. The effects of depression and severity of dementia on the mortality rates seem to be largely independent. PMID- 10473326 TI - An assessment of the Standardized Assessment of Personality as a screening instrument for the International Personality Disorder Examination: a comparison of informant and patient assessment for personality disorder. AB - BACKGROUND: The International Personality Disorder Examination (IPDE) has been developed as a standardized interview for personality disorders. While it has good psychometric properties, its length makes it difficult to use in the community in population research, particularly outside psychiatric settings. The informant-based Standard Assessment of Personality (SAP), which has been in use since 1981, could serve as a valid screen to detect likely personality disordered individuals who would then receive a definitive diagnosis by IPDE. This study aimed to compare the two instruments in their capacity to detect personality disorder according to ICD-10 taxonomy and to estimate the efficiency of the use of the two together in a case-finding exercise. METHOD: Ninety psychiatric out patients in Bangalore, India, were assessed for personality disorder using the two methods. Assessment was conducted by a pair of trained interviewers in random order and by random allocation to interviewer. RESULTS: Overall agreement between the two instruments in the detection of ICD-10 personality disorder was modest (kappa = 0.4). The level of agreement varied according to personality category, ranging from kappa 0.66 (dependent) to kappa 0.09 (dyssocial). The SAP proved to have a high negative predictive value (97%) for IPDE as the gold standard, suggesting its potential as a screen in samples where the expected prevalence of personality disorder is low. CONCLUSION: A two-stage approach to epidemiological studies of personality disorder may be practicable. PMID- 10473327 TI - Mother-infant interaction in post-partum women with schizophrenia and affective disorders. AB - BACKGROUND: Psychiatric mother and baby units are increasingly asked to assess parenting in people with severe mental illness, particularly schizophrenia, but little research evidence exists on which to base assessments. METHOD: Mother infant interaction was assessed in 26 women who had recovered from the acute phase of severe post-partum mental disorder, a validated rating scale based on direct observation was used. RESULTS: Women with schizophrenia showed greater interaction deficits than those with affective disorders, being more remote, insensitive, intrusive and self-absorbed. The 4-month-old infants of women with schizophrenia were more avoidant, and the overall quality of mother-infant interaction in schizophrenia was poorer. CONCLUSION: The long-term significance of these preliminary findings is not known but they raise concerns about the parenting capacity of women with schizophrenia and suggest the need for an intervention to improve parenting skills in this group. PMID- 10473328 TI - Reduced activation and altered laterality in two neuroleptic-naive catatonic patients during a motor task in functional MRI. AB - BACKGROUND: Catatonia, a symptom complex with motor, affective and cognitive symptoms seen in a variety of psychotic conditions and with organic disease, was examined using a motor task using functional magnetic resonance imaging (fMRI). METHODS: Two acute catatonic patients and two age- and sex-matched healthy controls performed sequential finger opposition (SFO) after being medicated with 2 mg of lorazepam (i.v.). Functional magnetic resonance images were collected using a gradient echo pulse sequence (EPI). RESULTS: Patients with catatonia showed reduced motor activation of the contralateral motor cortex during SFO of the right hand, ipsilateral activation was similar for patients and controls. There were no differences in the activation of the SMA. During left hand activation the right-handed catatonic patients showed more activation in the ipsilateral cortex, a reversal from the normal pattern of activation in which the contralateral side shows four to five times more activation than the ipsilateral side. CONCLUSIONS: In catatonic patients there is a decreased activation in motor cortex during a motor task compared to matched medicated healthy controls. In addition activation of the non-dominant side, left-handed activity in right handed patients, results in a total reversal of the normal pattern of lateral activation suggesting a disturbance in hemispheric localization of activity during a catatonic state. PMID- 10473329 TI - Randomizing carotid endarterectomy to carotid stenting? PMID- 10473330 TI - Endovascular repair of abdominal aortic aneurysm using the EVT device: limited increased utilization with availability of a bifurcated graft. AB - PURPOSE: To determine if the availability of a bifurcated graft would increase the percentage of patients eligible for endovascular repair of abdominal aortic aneurysms (AAAs). METHODS: One hundred eighty-five consecutive patients were evaluated prospectively for endovascular AAA repair at a university referral center. Data were collected on eligibility for tube or bifurcated endovascular grafts, reasons for exclusion, aneurysm morphology, and the interventions performed. RESULTS: Forty-six (25%) patients were eligible for endovascular treatment using the first-generation Endovascular Technologies (EVT) system: 19 (10%) for a tube graft and 27 (15%) for a bifurcated device. An unsuitable proximal neck was the reason for exclusion in 48% of patients (excess diameter in 27%, inadequate length in 21%). Unsuitable iliac configuration was present in 41% of those excluded; 29% of the common iliac arteries were enlarged or aneurysmal, while 12% were small or tortuous. CONCLUSIONS: Although a bifurcated graft more than doubles the eligibility of AAA patients for endovascular repair, the configuration of the proximal neck and iliac disease excluded the majority of AAA patients from endovascular therapy using the first generation EVT device. PMID- 10473331 TI - Endovascular repair of aortic aneurysms: treatment of complications. AB - PURPOSE: To evaluate the use of interventional procedures for treating complications following endovascular repair of aortic aneurysms. METHODS: Fifty five patients (49 men; mean age 67.5 years) underwent endoluminal stent-graft repair of traumatic (n = 4) or arteriosclerotic (n = 51) aortic aneurysms in the thoracic (n = 3) or infrarenal (n = 52) aorta. Follow-up of therapeutic success included periodic clinical examination, angiography, and spiral computed tomography. RESULTS: Discounting the 25 (45%) cases of postimplantation syndrome that did not require treatment, there were 22 complications observed in 20 (36%) patients over a mean 10-month follow-up (range 1 to 27). There were 2 transrenal endograft maldeployments, 1 case of twisted graft limbs, 2 access site problems (1 patient), 12 endoleaks (11 patients), 1 late graft limb thrombosis, 1 symptomatic internal iliac artery occlusion, 2 myocardial infarctions, and 1 transient psychosis. Seven (13%) patients did not undergo specific therapy, while 4 (7%) required operation (2 crossover bypass grafts, 1 suture revision, and 1 graft replacement). Among 9 (16%) patients treated with interventional techniques, 7 underwent percutaneous coil embolization for 8 endoleaks (7 successfully resolved). One late stent-graft disconnection required an additional stent-graft, and 1 of the 2 malpositioned endografts was repositioned. All patients remain alive with no increase in the diameter of the aneurysm in any patient. CONCLUSIONS: Technical problems resulting from the endovascular repair of aortic aneurysms often respond to interventional treatment. PMID- 10473332 TI - Subfascial perforator vein ablation: comparison of open versus endoscopic techniques. AB - PURPOSE: To compare the outcomes and complications of open (OSPS) versus endoscopic subfascial perforator surgery (SEPS) for treatment of chronic venous insufficiency. METHODS: Data were retrospectively collected on 25 patients who underwent 27 SEPSs from February 1996 to August 1997 and from 22 patients who underwent 29 OSPSs between March 1978 and May 1993. Outcomes were evaluated for postoperative complications, ulcer healing, recurrence, and venous dysfunction scores on the last follow-up for the SEPS group and at 1-year follow-up for the OSPS group. RESULTS: The 2 groups were similar in age, sex, history of previous venous surgery, healed or active ulcers, etiology, deep venous incompetency, pathophysiology, and venous refill times. Eighteen (90%) of 20 active ulcers in the SEPS group healed with recurrences in 5 (28%) limbs at 7.5 +/- 5.4-month follow-up. All 19 ulcers in the OSPS group healed, with recurrences in 13 (68%) limbs at 35 +/- 35-month follow-up. Clinical venous dysfunction scores showed significant improvement following SEPS (10.0 +/- 3.6 to 5.4 +/- 4.1, p < 0.001) and OSPS (10.0 +/- 3.2 to 6.7 +/- 3.6, p < 0.001) with no significant difference between groups. Both groups also had significant improvement in anatomical and disability scores. There was no postoperative mortality in either group. The OSPS group had significantly more wound complications (45%) than the SEPS group (7%) (p < 0.005). The hospital stay and readmission rate for wound problems were also higher in the OSPS group. CONCLUSIONS: The early outcome showed equal improvement in clinical venous dysfunction scores in the 2 groups, but with significantly fewer complications in the SEPS group. Although the long-term durability of the endoscopic approach has not been determined, the short-term results would favor SEPS for treatment of severe venous insufficiency when perforator incompetence is a significant component. PMID- 10473333 TI - Percutaneous stenting of the internal carotid artery: the European CAST I Study. Carotid Artery Stent Trial. AB - PURPOSE: To report the results of a multicenter safety trial of percutaneous carotid stenting performed by vascular surgeons. METHODS: Symptomatic or asymptomatic patients > or = 65 years of age with internal carotid artery (ICA) stenoses > or = 70% and < or = 2-cm long were eligible for enrollment. The procedures were performed in an operating room with the choice of anesthesia and the percutaneous access site at the discretion of the surgeon. Only Palmaz stents were used. RESULTS: From January 1, 1996 to December 31, 1997, 99 patients (74 men, mean age 70 years, range 51 to 94) were enrolled in the study. More than half (57 of 99 patients) were asymptomatic. The direct cervical approach was used predominantly (97%). Three (3%) cases were converted to surgery for inability to access the artery or deploy the stent (technical success 97%). No perioperative death or myocardial infarction was reported. Six (6%) procedural complications included 1 reversible arterial spasm, 2 dissections, 1 cervical hematoma, and 2 residual stenoses. One neurological event reversed within 7 days (1% minor stroke rate) and 4 (4%) transient ischemic attacks resolved within 24 hours. One (1%) asymptomatic early occlusion occurred 2 days postoperatively. No neurological event was observed in the 1- to 24-month follow-up (mean 13 months). Two (2%) patients died of nonprocedurally related causes. No stent compression was seen, but 1 asymptomatic occlusion and 3 asymptomatic, non-flow-limiting restenoses (2 < 40%, 1 at 60%) were found within 1 year (3% restenosis rate on an intention-to treat basis). Patency was 98% at 1 year. CONCLUSIONS: The results of this trial support the contention that carotid stenting of short ICA lesions can be performed with a low neurological complication rate. PMID- 10473335 TI - Closure of large percutaneous access sites using the Prostar XL Percutaneous Vascular Surgery device. AB - PURPOSE: To report early experience using a vascular closure device following endovascular aortic aneurysmal repair in which large-bore sheaths are used. TECHNIQUE: A 10F Prostar XL Percutaneous Vascular Surgery device is used to deploy sutures around sheath entry sites up to 16F. At the completion of the procedure, the sutures are tied with a sliding knot to ensure adequate hemostasis. CONCLUSIONS: Maintaining the minimal invasiveness of the percutaneous approach to aortic endografting reduces patient discomfort and permits earlier ambulation and hospital discharge. Initial success seems to be maintained at 1 month, however, longer follow-up will be required. PMID- 10473334 TI - Ambulatory endovascular surgery: cost advantage and factors influencing its safe performance. AB - PURPOSE: To perform a retrospective analysis of chronic limb ischemia and determine whether endovascular surgery can be performed safely and cost effectively on an ambulatory basis. METHODS: Among 42 patients undergoing endovascular interventions for lower limb ischemia over a 12-month period ending October 1997, 32 (18 men, mean age 68 years, range 44 to 89) were treated solely by endovascular interventions. These patients were grouped according to time inhospital: 20 (63%) patients had endovascular procedures performed on an ambulatory basis, 7 (22%) were hospitalized for > 24 hours, and 5 (16%) required an overnight stay. The angioplasty procedures, which included placement of 21 stents, were performed in the abdominal aorta (n = 1) and the common iliac (n = 9), external iliac (n =7), superficial femoral (n = 11), popliteal (n = 5), tibioperoneal (n = 7), and subclavian (n = 1) arteries. Hospital charges were compared for the 3 groups. RESULTS: Other than the presence of coronary artery disease, there were no significant differences in demographics or risk factors among the 3 groups. Angioplasty was technically successful in all patients, and there were no procedural complications. Patients with tissue loss required hospitalization more frequently compared to those with claudication. Significantly more patients who were hospitalized had epidural anesthesia as opposed to local when compared to the ambulatory group, 43% versus 5%, respectively (p = 0.04). Excluding professional fees, mean total hospital cost differed significantly between the ambulatory group and the group of patients with a hospital stay > 24 hours ($8227 versus $40,383, respectively; p = 0.03) and between the 2 hospitalized groups ($9476 for overnight stay versus $40,383 for > 24-hour stay, p = 0.03). CONCLUSIONS: Peripheral endovascular interventions can be performed safely on an ambulatory basis resulting in decreased hospital cost. Patients who receive epidural anesthesia, require concomitant open vascular reconstruction, present with tissue loss, or have unstable medical conditions are more likely to require hospitalization. PMID- 10473336 TI - Endoluminal repair of aneurysms containing ostia of essential branch arteries: an experimental model. AB - PURPOSE: To examine experimentally the feasibility of transfemoral endoluminal repair of aneurysms containing the ostia of essential branch arteries. METHODS: In a canine model (n = 4), suprarenal aortic aneurysms were created by suturing an artificial patch onto an anterior arteriotomy. Following a 2-week recovery period, the dogs underwent endovascular exclusion of their aneurysms using an aortic stent-graft with separate renal artery branch grafts. Outcome was evaluated using angiography, intravascular ultrasound (IVUS), Doppler flow, invasive pressure monitoring, and autopsy, respectively. RESULTS: Successful creation and subsequent endovascular exclusion of the aneurysm using aortic stent grafts and separate bilateral renal artery stent-grafts was achieved in all trials. Angiographically, all aneurysms were excluded from aortic flow and all renal arteries were patent at completion of the procedure. With IVUS, good graft apposition and absence of perigraft flow were demonstrated in all animals. Mean pressure in the aneurysmal sac at completion of the procedure was 40 +/- 7 mmHg, compared to a mean systemic blood pressure of 105 +/- 8 mmHg (p < 0.05). At autopsy, no gross intimal damage was seen in the aorta or the renal arteries, and intact aortic grafts and branch grafts without twisting, coiling, or kinking were found in all trials. CONCLUSIONS: In an acute animal model, suprarenal aortic aneurysms can be excluded from the circulation with preservation of renal flow using an endoluminally placed aortic stent-graft with separate branch grafts. PMID- 10473337 TI - People with mental retardation as witnesses in court: a review. AB - Evidence concerning eyewitness testimony given by people with mental retardation in court was reviewed. Despite general perceptions that people with mental retardation make incompetent witnesses, available evidence suggests that they can provide accurate accounts of witnessed events. The accounts are usually less complete than those provided by the general population and are greatly influenced by the methods of questioning. The sparse available evidence suggests that cross examination methods may lead to memory distortion. The use of closed, complex, and leading questions and the absence of aids to recall may have a particularly adverse effect on people with mental retardation. Resulting errors could lead to a false conviction or acquittal. Future policy and research in this much neglected area were discussed. PMID- 10473338 TI - Protecting the social body: use of the organism metaphor in fighting the "menace of the feebleminded". AB - Although persons with developmental disabilities living in the United States have been treated in a pejorative manner at various times throughout the nation's history, the eugenics era (1900-1930) stands out as a time when such individuals then referred to as "feebleminded"--were subject to particularly extreme indignities. Numerous methods of dehumanizing such persons were employed during this era. Of special significance was the use of the organism metaphor, whereby the "unfit" members of society were compared to a parasite, cancer, virus, or plague infecting the social body. The use of rhetoric advancing the organism metaphor in eugenic writing is described in this paper as is the effect that such rhetoric had on the societal response to such persons. PMID- 10473339 TI - Treatment of adolescent sex offenders with intellectual disabilities. AB - The problems of male adolescent sex offenders with intellectual disabilities were described and issues for treatment reviewed. A group treatment based on cognitive therapy was offered. Treatment methods and assessment of attitudes related to commission of sexual offenses were described. Four case studies of teenage male adolescent sex offenders with intellectual disabilities were presented. All subjects responded to treatment. Their individual differences related to their responses were discussed in the context of the type of denial exhibited by each subject. At the time of this study, 3 years had elapsed for 2 subjects and 4 years for the other 2 without a recurrence of an offense. PMID- 10473340 TI - Competence to confess: measuring understanding and suggestibility of defendants with mental retardation. AB - Groups of individuals with or without mental retardation, all of whom were currently on probation, were tested with measures of comprehension of Miranda rights and of suggestibility. As in previous work, participants with mental retardation were found to be less able to comprehend their Miranda rights. In addition, they were significantly more likely to respond to suggestive questioning and to change their answers. Significant correlations were found between measures of comprehension of Miranda rights and measures of suggestibility. Implications of these results were discussed. PMID- 10473341 TI - "Best friends": the construction of a teenage friendship. AB - A case study of a friendship between two high school students with disabilities was reported. The study was part of a research project on the social relationships of children and youth with disabilities. Ethnographic interviews and participant observations were used to accomplish two goals: (a) delineating the features of a spontaneous friendship between two young women in a self contained high school class and (b) assisting them in pursuing social activities together outside of school. The "insider perspectives" of the participants regarding their friendship is presented as is an interpretation of factors that supported and constrained the relationship. The importance of professionals encouraging the formation of networks among the families of youth with disabilities and of respecting families' cultural perspectives is emphasized. PMID- 10473342 TI - Ambiguity of the external genitalia in an infant with Down syndrome: gender assignment and ethical implications. PMID- 10473343 TI - Historical perspective on mortality and placement. PMID- 10473344 TI - Spilled religion: the tragedy of Henry H. Goddard. PMID- 10473345 TI - Community residential services would require expansion of 72% to serve everyone in community settings. PMID- 10473346 TI - Replication protein A (RPA): the eukaryotic SSB. AB - Replication protein A (RPA) is a heterotrimeric single-stranded DNA-binding protein that is highly conserved in eukaryotes. RPA plays essential roles in many aspects of nucleic acid metabolism, including DNA replication, nucleotide excision repair, and homologous recombination. In this review, we provide a comprehensive overview of RPA structure and function and highlight the more recent developments in these areas. The last few years have seen major advances in our understanding of the mechanism of RPA binding to DNA, including the structural characterization of the primary DNA-binding domains (DBD) and the identification of two secondary DBDs. Moreover, evidence indicates that RPA utilizes a multistep pathway to bind single-stranded DNA involving a particular molecular polarity of RPA, a mechanism that is apparently used to facilitate origin denaturation. In addition to its mechanistic roles, RPA interacts with many key factors in nucleic acid metabolism, and we discuss the critical nature of many of these interactions to DNA metabolism. RPA is a phosphorylation target for DNA-dependent protein kinase (DNA-PK) and likely the ataxia telangiectasia mutated gene (ATM) protein kinase, and recent observations are described that suggest that RPA phosphorylation plays a significant modulatory role in the cellular response to DNA damage. PMID- 10473347 TI - Molecular elements of ion permeation and selectivity within calcium channels. AB - Voltage-dependent calcium channels are located in the plasma membrane and form a highly selective conduit by which Ca2+ ions enter all excitable cells and some nonexcitable cells. Extensive characterization studies have revealed the existence of one low (T) and five high-voltage-activated calcium channel types (L, N, P, Q, and R). The high voltage-activated calcium channels have been found to exist as heteromultimers, consisting of an alpha1, beta, alpha2/delta, and gamma subunit. Molecular cloning has revealed the existence of 10 channel transcripts, and expression of these cloned calcium channel genes has shown that basic voltage-activated calcium channel function is strictly carried by the corresponding alpha1 subunits. In turn, the auxiliary subunits serve to modulate calcium channel function by altering the voltage dependence of channel gating, kinetics, and current amplitude, thereby creating a likelihood for calcium channels with multiple properties. Although for calcium channels to be effective, Ca2+ ions must enter selectively through the pore of the alpha1-subunit, bypassing competition with other extracellular ions. The structural determinants of this highly selective Ca2+ filter reside within the four glutamic acid residues located at homologous positions within each of the four pore-forming segments. Together, these residues form a single or multiple Ca2+ affinity site(s) that entrap calcium ions, which are then electrostatically repulsed through the intracellular opening of the pore. This mechanism of high-selectivity calcium filtration, the spatial arrangement of pore glutamic acid residues, and the coordination chemistry of calcium binding are discussed in this review. PMID- 10473348 TI - Presentation of the Mayo brothers painting. PMID- 10473349 TI - NASA/DARPA advanced communications technology satellite project for evaluation of telemedicine outreach using next-generation communications satellite technology: Mayo Foundation participation. AB - OBJECTIVE: To describe the development of telemedicine capabilities-application of remote consultation and diagnostic techniques-and to evaluate the feasibility and practicality of such clinical outreach to rural and underserved communities with limited telecommunications infrastructures. MATERIAL AND METHODS: In 1992, Mayo Foundation (Rochester, Minn, Jacksonville, Fla, and Scottsdale, Ariz), the National Aeronautics and Space Administration, and the Defense Advanced Research Projects Agency collaborated to create a complex network of fiberoptic landlines, video recording systems, satellite terminals, and specially developed data translators linking Mayo sites with other locations in the continental United States on an on-demand basis. The purpose was to transmit data via the asynchronous transfer mode (ATM) digital communications protocol over the Advanced Communications Technology Satellite. The links were intended to provide a conduit for transmission of data for patient-specific consultations between physicians, evaluation of medical imagery, and medical education for clinical staffs at remote sites. RESULTS: Low-data-rate (LDR) experiments went live late in 1993. Mayo Clinic Rochester successfully provided medical consultation and services to 2 small regional medical facilities. High-data-rate (HDR) experiments included studies of remote digital echocardiography, store-and-forward telemedicine, cardiac catheterization, and teleconsultation for congenital heart disease. These studies combined landline data transmission with use of the satellite. The complexity of the routing paths and network components, immaturity of available software, and inexperience with existing telecommunications caused significant study delays. CONCLUSIONS: These experiments demonstrated that next generation satellite technology can provide batch and real-time imagery for telemedicine. The first-generation of the ATM and satellite network technology used in these experiments created several technical problems and inconveniences that should be overcome as the network infrastructure matures. PMID- 10473350 TI - Telemedicine consultations in congenital heart disease: assessment of advanced technical capabilities. AB - OBJECTIVE: To assess the value of adding remote pointer and dynamic display capabilities to a telemedicine system designed to provide consultative services for patients with congenital heart disease. MATERIAL AND METHODS: Independent observations by the referring physician and the consulting physician provided the data for the assessment. Fifty-four teleconsultations involving 38 patients with 21 different congenital heart diseases were analyzed. The teleconsultations were based on previously obtained cineangiograms that were digitized and then transmitted by combined satellite and terrestrial-based technology. The observations, recorded by each physician at his workstation at the time of each teleconsultation, were summarized and analyzed statistically. RESULTS: In 108 observations, the pointer was believed to be helpful in 72 (67%), and dynamic display was helpful in 96 (89%). CONCLUSION: This study suggests that use of a pointer and dynamic display enhances teleconsultations for patients with congenital heart disease. PMID- 10473351 TI - Clinical consultations using store-and-forward telemedicine technology. AB - OBJECTIVE: To investigate the potential role of a store-and-forward (SAF) telemedicine system in specialty consultations initiated by primary care physicians. MATERIALS AND METHODS: In this pilot telemedicine study, patients needing consultations in cardiology, dermatology, endocrinology, and orthopedics had both standard face-to-face (FTF) consultations and SAF consultations. RESULTS: Fifteen patients had both FTF and SAF consultations, 4 had echocardiograms transmitted for an SAF consultation only, and 1 had an SAF consultation but no FTF appointment. Of 19 diagnoses made, all were essentially the same in both types of consultations; 14 of 15 FTF consultations and 15 of 19 SAF consultations resulted in additional treatment recommendations. CONCLUSIONS: While it was possible to develop a desktop system for SAF consultations, the equipment was not adequately integrated. Without total digital input, including electronic patient medical records, packaging of information is laborious and impractical. Seamlessly adapting to existing clinical practice is vital. Issues such as increasing work for the physicians or office staff, gathering adequate patient information, and designing a referral process were more difficult than we had anticipated. Patient acceptance was high, but the clinical pilot had very small numbers. PMID- 10473352 TI - Self-perception in elite collegiate female gymnasts, cross-country runners, and track-and-field athletes. AB - OBJECTIVE: To compare self-perception between a group of competitive, elite female collegiate athletes (participating in gymnastics, cross country, and track and field) and a group of female political science students (nonathletic control subjects). We hypothesized that the athletic group would rate athletics as more important than would the nonathletic group, that the perception of athletic competence would correlate positively with self-worth for athletes only, and that the perception of athletic competence would have a stronger influence on self worth in the athletic group. SUBJECTS AND METHODS: The Self-perception Profile for College Students was completed by 32 athletes and 13 nonathletes. This profile measures 12 subscales plus Global Self-worth independently and generates scores that reflect the subject's perceived importance of and competence in each of the subscale areas. RESULTS: The athletes rated athletics as more important than did nonathletes, although this trend was nonsignificant when adjusted for age. As age increased, the importance of athletics decreased for both groups. There was a direct relationship between perceived athletic competence and self worth for the athletes but not for the nonathletes. Variables that accounted for the Global Self-worth score in athletes were perceptions of Competence subscales for Appearance, Social Acceptance, Friendship, and Job. Variables that accounted for the Global Self-worth scores in the nonathletes were perceptions of Competence subscales for Romance, Morality, Humor, and Appearance. The athletic group had significantly lower Global Self-worth scores than the nonathletic group. CONCLUSIONS: The female athletes in this study derived a large component of their self-worth from their perceived athletic competence. Clinicians should bear in mind the relative importance of athletics to young female athletes and the relationship of perceived athletic ability to self-worth when treating these individuals. PMID- 10473353 TI - Intracardiac echocardiography via the transvenous approach with use of 8F 10-MHz ultrasound catheters. AB - OBJECTIVE: To study the safety and feasibility of intracardiac imaging using a novel 8F 10-MHz non-over-the-wire ultrasound catheter system. SUBJECTS AND METHOD: Intracardiac imaging using a transfemoral venous approach was performed in 33 adults, 14 men and 19 women, aged 25 to 66 years (mean, 46 years). Six were normal subjects, 12 had congenital heart diseases (5 atrial septal defects, 3 ventricular septal defects, 1 tetralogy of Fallot, 2 patent ductus arteriosus, and 1 Ebstein anomaly), 14 had valvular heart diseases (12 mitral stenoses and 2 calcific aortic stenoses), and 1 had acute pulmonary embolism. RESULTS: Ultrasound images were obtained, without any complications, from the right side of the heart in all subjects. The atria and ventricles could be recognized by anatomic relationships to the cardiac chambers and the valves by their characteristic motion during each cardiac cycle. The vessels were verified by their connections to the cardiac chamber and by contrast echocardiography if indicated. CONCLUSION: Intracardiac imaging using the 8F 10-MHz non-over-the-wire ultrasound catheter system via a transfemoral venous approach is feasible and safe. Intracardiac echocardiography is potentially useful for assessing a variety of cardiac anomalies and in guiding and monitoring certain intervention procedures. PMID- 10473354 TI - Bert Sakmann--Nobel Prize for studies of cell function. PMID- 10473355 TI - A commonsense approach to shoulder problems. AB - Shoulder pain is a common entity in a primary care physician's practice. The unique anatomy of the shoulder allows for almost unrestrained motion in all planes. A thorough history and physical examination are important to ensure efficient patient evaluation. Further assessment may include radiographic and diagnostic laboratory tests. This article presents an organized approach to the anatomy, physiology, and pathology of common shoulder disorders for the primary care physician. The distinction between disorders that are intrinsic or extrinsic to the shoulder joint is discussed. Treatment and the need for appropriate referral are described. PMID- 10473356 TI - Bilateral thalamic infarction in a patient with factor V Leiden mutation. AB - In a 50-year-old unresponsive man with ophthalmoplegia, bilateral thalamic infarction was detected on magnetic resonance imaging of the brain. A thorough evaluation, including urinalysis, laboratory studies, computed tomography of the brain, chest radiography, lumbar puncture, magnetic resonance angiography, cerebral angiography, carotid ultrasonography, and transesophageal echocardiography, revealed no obvious predisposing factors other than heterozygous factor V Leiden mutation. To our knowledge, this is the first reported case of bilateral thalamic infarction associated with factor V Leiden mutation. Physicians should consider the possibility of this mutation in patients with ischemic stroke if no other source is evident. PMID- 10473357 TI - Multiple cerebral infarctions from nonbacterial thrombotic endocarditis mimicking cerebral vasculitis. AB - Primary vasculitis of the central nervous system (PVCNS) is an uncommon disorder that can present with a variety of symptoms, making diagnosis and management difficult. We describe a case of cerebral infarction that occurred from nonbacterial thrombotic endocarditis (NBTE) and presented with clinical and radiologic imaging features that suggested PVCNS. The patient was a 58-year-old woman with left hemiparesis, aphasia, and episodic confusion. Magnetic resonance imaging of the brain demonstrated multifocal lesions consistent with infarction involving both cerebral hemispheres, and cerebral angiography showed changes consistent with vasculitis. Although brain biopsy findings were normal, the patient was treated for presumed vasculitis with cyclophosphamide and prednisone. Four months later respiratory failure secondary to polymicrobial pneumonia and adult respiratory distress syndrome developed, and she died. Autopsy revealed multiple infarcts in the heart, lungs, right kidney, spleen, and brain. Multiple thrombotic platelet-fibrin vegetations consistent with NBTE were found on all cardiac valves. Examination of the brain revealed no evidence of active or healed vasculitis. Cerebral angiography may show findings that suggest vasculitis, but it is not diagnostic, as several other conditions may cause similar changes. Nonbacterial thrombotic endocarditis may cause multiple cerebral infarctions and can be difficult to distinguish from vasculitis, as specific diagnostic tests for PVCNS are lacking. PMID- 10473358 TI - Central nervous system histoplasmosis mimicking a brain tumor: difficulties in diagnosis and treatment. AB - Neurologic involvement occurs in 10% to 20% of patients with disseminated histoplasmosis. We describe a 20-year-old woman who had headache and diplopia but no evidence of systemic infection. Magnetic resonance imaging showed an enhancing mass in the thalamomesencephalic and third ventricular region. After subtotal resection of what was presumed to be a glioma, the patient had symptoms and signs of meningitis. Subsequent pathological review demonstrated noncaseating granulomas, and serologic tests and cultures confirmed the diagnosis of histoplasmosis. Initiation of antifungal therapy and removal of an infected shunt system resulted in clinical improvement. Clinicians should maintain a high index of suspicion in patients who are from any area endemic for histoplasmosis. PMID- 10473359 TI - Current guidelines for the management of small cell lung cancer. AB - Small cell lung cancer (SCLC) accounts for 20% to 25% of cases of bronchogenic carcinoma and results in pronounced morbidity and mortality in the United States. More than 90% of cases of SCLC are caused by cigarette smoking. Common pulmonary manifestations are dyspnea, persistent cough, hemoptysis, and postobstructive pneumonia. At the time of diagnosis, patients usually have extensive disease. To date, therapeutic approaches have made only modest advances in outcome. Combined modality approaches, such as radiotherapy administered concomitantly with the initiation of chemotherapy, induction chemotherapy followed by radiotherapy administered during the subsequent courses of chemotherapy, sequential chemotherapy and radiotherapy, and courses of radiotherapy split between cycles of chemotherapy, are important for improving survival in patients with SCLC. PMID- 10473360 TI - Noninvasive ventilation. AB - Noninvasive ventilation refers to the delivery of assisted ventilatory support without the use of an endotracheal tube. Noninvasive positive pressure ventilation (NPPV) can be delivered by using a volume-controlled ventilator, a pressure-controlled ventilator, a bilevel positive airway pressure ventilator, or a continuous positive airway pressure device. During the past decade, there has been a resurgence in the use of noninvasive ventilation, fueled by advances in technology and clinical trials evaluating its use. Several manufacturers produce portable devices that are simple to operate. This review describes the equipment, techniques, and complications associated with NPPV and also the indications for both short-term and long-term applications. NPPV clearly represents an important addition to the techniques available to manage patients with respiratory failure. Future clinical trials evaluating its many clinical applications will help to define populations of patients most apt to benefit from this type of treatment. PMID- 10473361 TI - 49-year-old woman with chest pain. PMID- 10473362 TI - Clindamycin, metronidazole, and chloramphenicol. AB - Clindamycin, metronidazole, and chloramphenicol are three antimicrobial agents useful in the treatment of anaerobic infections. Clindamycin is effective in the treatment of most infections involving anaerobes and gram-positive cocci, but emerging resistance has become a problem in some clinical settings. Metronidazole is effective in the treatment of infections involving gram-negative anaerobes, but it is unreliable in the treatment of gram-positive anaerobic infections and is ineffective in treating aerobic infections. Additionally, metronidazole is often the drug of choice in treating infections in which Bacteroides fragilis is a serious concern. Chloramphenicol is effective in the treatment of a wide variety of bacterial infections, including serious anaerobic infections, but is rarely used in Western countries because of concerns about toxicity, including aplastic anemia and gray baby syndrome. PMID- 10473363 TI - Pathways to primary neurodegenerative disease. PMID- 10473364 TI - Eliminating disparities, promoting partnerships. PMID- 10473365 TI - Technology, sociology, and medicine. PMID- 10473366 TI - Tattoos, body piercing and thrush: a lesson on the harmful effects of lost objectivity. PMID- 10473367 TI - Humility and the practice of medicine. PMID- 10473368 TI - Pulmonary hypertension in systemic lupus erythematosus. PMID- 10473369 TI - Silent but widespread use of dietary supplements. PMID- 10473370 TI - First human vaccine for Lyme disease. PMID- 10473371 TI - Fibrin meshwork as a carrier for delivery of angiogenic growth factors in patients with ischemic limb. PMID- 10473372 TI - The scientific basis for probiotic strains of Lactobacillus. PMID- 10473374 TI - Role of Ngamma-acetyldiaminobutyrate as an enzyme stabilizer and an intermediate in the biosynthesis of hydroxyectoine. AB - Strain CHR63 is a salt-sensitive mutant of the moderately halophilic wild-type strain Halomonas elongata DSM 3043 that is affected in the ectoine synthase gene (ectC). This strain accumulates large amounts of Ngamma-acetyldiaminobutyrate (NADA), the precursor of ectoine (D. Canovas, C. Vargas, F. Iglesias-Guerra, L. N. Csonka, D. Rhodes, A. Ventosa, and J. J. Nieto, J. Biol. Chem. 272:25794 25801, 1997). Hydroxyectoine, ectoine, and glucosylglycerate were also identified by nuclear magnetic resonance (NMR) as cytoplasmic organic solutes in this mutant. Accumulation of NADA, hydroxyectoine, and ectoine was osmoregulated, whereas the levels of glucosylglycerate decreased at higher salinities. The effect of the growth stage on the accumulation of solutes was also investigated. NADA was purified from strain CHR63 and was shown to protect the thermolabile enzyme rabbit muscle lactate dehydrogenase against thermal inactivation. The stabilizing effect of NADA was greater than the stabilizing effect of ectoine or potassium diaminobutyrate. A (1)H NMR analysis of the solutes accumulated by the wild-type strain and mutants CHR62 (ectA::Tn1732) and CHR63 (ectC::Tn1732) indicated that H. elongata can synthesize hydroxyectoine by two different pathways-directly from ectoine or via an alternative pathway that converts NADA into hydroxyectoine without the involvement of ectoine. PMID- 10473375 TI - Polyphosphate kinase of Acinetobacter sp. strain ADP1: purification and characterization of the enzyme and its role during changes in extracellular phosphate levels. AB - Polyphosphate (polyP) is a ubiquitous biopolymer whose function and metabolism are incompletely understood. The polyphosphate kinase (PPK) of Acinetobacter sp. strain ADP1, an organism that accumulates large amounts of polyP, was purified to homogeneity and characterized. This enzyme, which adds the terminal phosphate from ATP to a growing chain of polyP, is a 79-kDa monomer. PPK is sensitive to magnesium concentrations, and optimum activity occurs in the presence of 3 mM MgCl(2). The optimum pH was between pH 7 and 8, and significant reductions in activity occurred at lower pH values. The greatest activity occurred at 40 degrees C. The half-saturation ATP concentration for PPK was 1 mM, and the maximum PPK activity was 28 nmol of polyP monomers per microg of protein per min. PPK was the primary, although not the sole, enzyme responsible for the production of polyP in Acinetobacter sp. strain ADP1. Under low-phosphate (P(i)) conditions, despite strong induction of the ppk gene, there was a decline in net polyP synthesis activity and there were near-zero levels of polyP in Acinetobacter sp. strain ADP1. Once excess phosphate was added to the P(i)-starved culture, both the polyP synthesis activity and the levels of polyP rose sharply. Increases in polyP-degrading activity, which appeared to be mainly due to a polyphosphatase and not to PPK working in reverse, were detected in cultures grown under low-P(i) conditions. This activity declined when phosphate was added. PMID- 10473373 TI - Biocontrol of Escherichia coli O157 with O157-specific bacteriophages. AB - Escherichia coli O157 antigen-specific bacteriophages were isolated and tested to determine their ability to lyse laboratory cultures of Escherichia coli O157:H7. A total of 53 bovine or ovine fecal samples were enriched for phage, and 5 of these samples were found to contain lytic phages that grow on E. coli O157:H7. Three bacteriophages, designated KH1, KH4, and KH5, were evaluated. At 37 or 4 degrees C, a mixture of these three O157-specific phages lysed all of the E. coli O157 cultures tested and none of the non-O157 E. coli or non-E. coli cultures tested. These results required culture aeration and a high multiplicity of infection. Without aeration, complete lysis of the bacterial cells occurred only after 5 days of incubation and only at 4 degrees C. Phage infection and plaque formation were influenced by the nature of the host cell O157 lipopolysaccharide (LPS). Strains that did not express the O157 antigen or expressed a truncated LPS were not susceptible to plaque formation or lysis by phage. In addition, strains that expressed abundant mid-range-molecular-weight LPS did not support plaque formation but were lysed in liquid culture. Virulent O157 antigen-specific phages could play a role in biocontrol of E. coli O157:H7 in animals and fresh foods without compromising the viability of other normal flora or food quality. PMID- 10473376 TI - Development of an in vitro bioassay for Clostridium botulinum type B neurotoxin in foods that is more sensitive than the mouse bioassay. AB - A novel, in vitro bioassay for detection of the botulinum type B neurotoxin in a range of media was developed. The assay is amplified by the enzymic activity of the neurotoxin's light chain and includes the following three stages: first, a small, monoclonal antibody-based immunoaffinity column captures the toxin; second, a peptide substrate is cleaved by using the endopeptidase activity of the type B neurotoxin; and finally, a modified enzyme-linked immunoassay system detects the peptide cleavage products. The assay is highly specific for type B neurotoxin and is capable of detecting type B toxin at a concentration of 5 pg ml(-1) (0.5 mouse 50% lethal dose ml(-1)) in approximately 5 h. The format of the test was found to be suitable for detecting botulinum type B toxin in a range of foodstuffs with a sensitivity that exceeds the sensitivity of the mouse assay. Using highly specific monoclonal antibodies as the capture phase, we found that the endopeptidase assay was capable of differentiating between the type B neurotoxins produced by proteolytic and nonproteolytic strains of Clostridium botulinum type B. PMID- 10473377 TI - Development and characterization of a gene expression reporter system for Clostridium acetobutylicum ATCC 824. AB - A gene expression reporter system (pHT3) for Clostridium acetobutylicum ATCC 824 was developed by using the lacZ gene from Thermoanaerobacterium thermosulfurogenes EM1 as the reporter gene. In order to test the reporter system, promoters of three key metabolic pathway genes, ptb (coding for phosphotransbutyrylase), thl (coding for thiolase), and adc (coding for acetoacetate decarboxylase), were cloned upstream of the reporter gene in pHT3 in order to construct vectors pHT4, pHT5, and pHTA, respectively. Detection of beta galactosidase activity in time course studies performed with strains ATCC 824(pHT4), ATCC 824(pHT5), and ATCC 824(pHTA) demonstrated that the reporter gene produced a functional beta-galactosidase in C. acetobutylicum. In addition, time course studies revealed differences in the beta-galactosidase specific activity profiles of strains ATCC 824(pHT4), ATCC 824(pHT5), and ATCC 824(pHTA), suggesting that the reporter system developed in this study is able to effectively distinguish between different promoters. The stability of the beta galactosidase produced by the reporter gene was also examined with strains ATCC 824(pHT4) and ATCC 824(pHT5) by using chloramphenicol treatment to inhibit protein synthesis. The data indicated that the beta-galactosidase produced by the lacZ gene from T. thermosulfurogenes EM1 was stable in the exponential phase of growth. In pH-controlled fermentations of ATCC 824(pHT4), the kinetics of beta galactosidase formation from the ptb promoter and phosphotransbutyrylase formation from its own autologous promoter were found to be similar. PMID- 10473378 TI - An effective strategy, applicable to Streptococcus salivarius and related bacteria, to enhance or confer electroporation competence. AB - Despite the large number of techniques available for transformation of bacteria, certain species and strains are still resistant to introduction of foreign DNA. Some oral streptococci are among the organisms that can be particularly difficult to transform. We performed a series of experiments that involved manipulation of growth and recovery media and cell wall weakening, in which the electroporation conditions, cell concentration, and type and concentration of the transforming plasmid were varied. The variables were optimized such that a previously untransformable Streptococcus salivarius strain, ATCC 25975, could be transformed reproducibly at a level of 10(5) transformants per microg of DNA. The technique was used to introduce a plasmid into other strains of S. salivarius, including a fresh isolate. Moreover, the same technique was applied successfully to a wide range of oral streptococci and other gram-positive bacteria. PMID- 10473379 TI - Polycyclic aromatic hydrocarbon metabolism by white rot fungi and oxidation by Coriolopsis gallica UAMH 8260 laccase. AB - We studied the metabolism of polycyclic aromatic hydrocarbons (PAHs) by using white rot fungi previously identified as organisms that metabolize polychlorinated biphenyls. Bran flakes medium, which has been shown to support production of high levels of laccase and manganese peroxidase, was used as the growth medium. Ten fungi grown for 5 days in this medium in the presence of anthracene, pyrene, or phenanthrene, each at a concentration of 5 microg/ml could metabolize these PAHs. We studied the oxidation of 10 PAHs by using laccase purified from Coriolopsis gallica. The reaction mixtures contained 20 microM PAH, 15% acetonitrile in 60 mM phosphate buffer (pH 6), 1 mM 2,2'-azinobis-(3 ethylbenzthiazoline-6-sulfonate) (ABTS), and 5 U of laccase. Laccase exhibited 91% of its maximum activity in the absence of acetonitrile. The following seven PAHs were oxidized by laccase: benzo[a]pyrene, 9-methylanthracene, 2 methylanthracene, anthracene, biphenylene, acenaphthene, and phenanthrene. There was no clear relationship between the ionization potential of the substrate and the first-order rate constant (k) for substrate loss in vitro in the presence of ABTS. The effects of mediating substrates were examined further by using anthracene as the substrate. Hydroxybenzotriazole (HBT) (1 mM) supported approximately one-half the anthracene oxidation rate (k = 2.4 h(-1)) that ABTS (1 mM) supported (k = 5.2 h(-1)), but 1 mM HBT plus 1 mM ABTS increased the oxidation rate ninefold compared with the oxidation rate in the presence of ABTS, to 45 h(-1). Laccase purified from Pleurotus ostreatus had an activity similar to that of C. gallica laccase with HBT alone, with ABTS alone, and with 1 mM HBT plus 1 mM ABTS. Mass spectra of products obtained from oxidation of anthracene and acenaphthene revealed that the dione derivatives of these compounds were present. PMID- 10473380 TI - Transformation of sulfur compounds by an abundant lineage of marine bacteria in the alpha-subclass of the class Proteobacteria. AB - Members of a group of marine bacteria that is numerically important in coastal seawater and sediments were characterized with respect to their ability to transform organic and inorganic sulfur compounds. Fifteen strains representing the Roseobacter group (a phylogenetic cluster of marine bacteria in the alpha subclass of the class Proteobacteria) were isolated from seawater, primarily from the southeastern United States. Although more than one-half of the isolates were obtained without any selection for sulfur metabolism, all of the isolates were able to degrade the sulfur-containing osmolyte dimethyl sulfoniopropionate (DMSP) with production of dimethyl sulfide (DMS). Five isolates also degraded DMSP with production of methanethiol, indicating that both cleavage and demethylation pathways for DMSP occurred in the same organism, which is unusual. Five isolates were able to reduce dimethyl sulfoxide to DMS, and several isolates also degraded DMS and methanethiol. Sulfite oxygenase activity and methanesulfonic acid oxygenase activity were also present in some of the isolates. The ability to incorporate the reduced sulfur in DMSP and methanethiol into cellular material was studied with one of the isolates. A group-specific 16S rRNA probe indicated that the relative abundance of uncultured bacteria in the Roseobacter group increased in seawater enriched with DMSP or DMS. Because this group typically accounts for >10% of the 16S ribosomal DNA pool in coastal seawater and sediments of the southern United States, clues about its potential biogeochemical role are of particular interest. Studies of culturable representatives suggested that the group could mediate a number of steps in the cycling of both organic and inorganic forms of sulfur in marine environments. PMID- 10473381 TI - Marine bacterial isolates display diverse responses to UV-B radiation. AB - The molecular and biological consequences of UV-B radiation were investigated by studying five species of marine bacteria and one enteric bacterium. Laboratory cultures were exposed to an artificial UV-B source and subjected to various post UV irradiation treatments. Significant differences in survival subsequent to UV-B radiation were observed among the isolates, as measured by culturable counts. UV B-induced DNA photodamage was investigated by using a highly specific radioimmunoassay to measure cyclobutane pyrimidine dimers (CPDs). The CPDs determined following UV-B exposure were comparable for all of the organisms except Sphingomonas sp. strain RB2256, a facultatively oligotrophic ultramicrobacterium. This organism exhibited little DNA damage and a high level of UV-B resistance. Physiological conditioning by growth phase and starvation did not change the UV-B sensitivity of marine bacteria. The rates of photoreactivation following exposure to UV-B were investigated by using different light sources (UV-A and cool white light). The rates of photoreactivation were greatest during UV-A exposure, although diverse responses were observed. The differences in sensitivity to UV-B radiation between strains were reduced after photoreactivation. The survival and CPD data obtained for Vibrio natriegens when we used two UV-B exposure periods interrupted by a repair period (photoreactivation plus dark repair) suggested that photoadaptation could occur. Our results revealed that there are wide variations in marine bacteria in their responses to UV radiation and subsequent repair strategies, suggesting that UV-B radiation may affect the microbial community structure in surface water. PMID- 10473382 TI - Amplified fragment length polymorphism fingerprinting of Pseudomonas strains from a poultry processing plant. AB - Molecular typing has been used previously to identify and trace dissemination of pathogenic and spoilage bacteria associated with food processing. Amplified fragment length polymorphism (AFLP) is a novel DNA fingerprinting technique which is considered highly reproducible and has high discriminatory power. This technique was used to fingerprint 88 Pseudomonas fluorescens and Pseudomonas putida strains that were previously isolated from plate counts of carcasses at six processing stages and various equipment surfaces and environmental sources of a poultry abattoir. Clustering of the AFLP patterns revealed a high level of diversity among the strains. Six clusters (clusters I through VI) were delineated at an arbitrary Dice coefficient level of 0.65; clusters III (31 strains) and IV (28 strains) were the largest clusters. More than one-half (52.3%) of the strains obtained from carcass samples, which may have represented the resident carcass population, grouped together in cluster III. By contrast, 43.2% of the strains from most of the equipment surfaces and environmental sources grouped together in cluster IV. In most cases, the clusters in which carcass strains from processing stages grouped corresponded to the clusters in which strains from the associated equipment surfaces and/or environmental sources were found. This provided evidence that there was cross-contamination between carcasses and the abattoir environment at the DNA level. The AFLP data also showed that strains were being disseminated from the beginning to the end of the poultry processing operation, since many strains associated with carcasses at the packaging stage were members of the same clusters as strains obtained from carcasses after the defeathering stage. PMID- 10473383 TI - Spatial heterogeneity of bacterial populations along an environmental gradient at a shallow submarine hydrothermal vent near Milos Island (Greece). AB - The spatial heterogeneity of bacterial populations at a shallow-water hydrothermal vent in the Aegean Sea close to the island of Milos (Greece) was examined at two different times by using acridine orange staining for total cell counts, cultivation-based techniques, and denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA gene fragments. Concurrent with measurements of geochemical parameters, samples were taken along a transect from the center of the vent to the surrounding area. Most-probable-number (MPN) counts of metabolically defined subpopulations generally constituted a minor fraction of the total cell counts; both counting procedures revealed the highest cell numbers in a transition zone from the strongly hydrothermally influenced sediments to normal sedimentary conditions. Total cell counts ranged from 3.2 x 10(5) cells ml(-1) in the water overlying the sediments to 6.4 x 10(8) cells g (wet weight) of sediment(-1). MPN counts of chemolithoautotrophic sulfur-oxidizing bacteria varied between undetectable and 1.4 x 10(6) cells g(-1). MPN counts for sulfate reducing bacteria and dissimilatory iron-reducing bacteria ranged from 8 to 1.4 x 10(5) cells g(-1) and from undetectable to 1.4 x 10(6) cells g(-1), respectively. DGGE revealed a trend from a diverse range of bacterial populations which were present in approximately equal abundance in the transition zone to a community dominated by few populations close to the center of the vent. Temperature was found to be an important parameter in determining this trend. However, at one sampling time this trend was not discernible, possibly due to storm-induced disturbance of the upper sediment layers. PMID- 10473384 TI - Distribution and diversity of sulfur-oxidizing Thiomicrospira spp. at a shallow water hydrothermal vent in the Aegean Sea (Milos, Greece). AB - A shallow-water hydrothermal vent system in the Aegean Sea close to the island of Milos (Greece) was chosen to study the diversity and distribution of the chemolithoautotrophic sulfur-oxidizing bacterium Thiomicrospira. Cell numbers in samples from different regions around a solitary vent decreased toward the center of the vent (horizontal distribution), as well as with depth (vertical distribution), corresponding to an increase in temperature (from ca. 25 to 60 degrees C) and a decrease in pH (from ca. pH 7 to 5). Thiomicrospira was one of the most abundant culturable sulfur oxidizers and was even dominant in one region. Phylogenetic analysis of Thiomicrospira spp. present in the highest most probable-number (MPN) dilutions revealed that most of the obtained sequences grouped in two new closely related clusters within the Thiomicrospira branch. Two different new isolates, i.e., Milos-T1 and Milos-T2, were obtained from high dilution (10(-5)) enrichments. Phylogenetic analysis indicated that isolate Milos T1 is related to the recently described Thiomicrospira kuenenii and Hydrogenovibrio marinus, whereas isolate Milos-T2 grouped with the MPN sequences of cluster 2. The predominance of strain Milos-T2 was indicated by its identification in several environmental samples by hybridization analysis of denaturing gradient gel electrophoresis (DGGE) patterns and by sequencing of one of the corresponding bands, i.e., ML-1, from the DGGE gel. The results shown in this paper support earlier indications that Thiomicrospira species are important members of hydrothermal vent communities. PMID- 10473385 TI - Development and use of a reverse transcription-PCR assay to study expression of Tri5 by Fusarium species in vitro and in planta. AB - The Tri5 gene encodes trichodiene synthase, which catalyzes the first reaction in the trichothecene biosynthetic pathway. In vitro, a direct relationship was observed between Tri5 expression and the increase in deoxynivalenol production over time. We developed a reverse transcription (RT)-PCR assay to quantify Tri5 gene expression in trichothecene-producing strains of Fusarium species. We observed an increase in Tri5 expression following treatment of Fusarium culmorum with fungicides, and we also observed an inverse relationship between Tri5 expression and biomass, as measured by beta-D-glucuronidase activity, during colonization of wheat (cv. Avalon) seedlings by F. culmorum. RT-PCR analysis also showed that for ears of wheat cv. Avalon inoculated with F. culmorum, there were different levels of Tri5 expression in grain and chaff at later growth stages. We used the Tri5-specific primers to develop a PCR assay to detect trichothecene producing Fusarium species in infected plant material. PMID- 10473386 TI - Transduction of enteric Escherichia coli isolates with a derivative of Shiga toxin 2-encoding bacteriophage phi3538 isolated from Escherichia coli O157:H7. AB - We investigated the ability of a detoxified derivative of a Shiga toxin 2 (Stx2) encoding bacteriophage to infect and lysogenize enteric Escherichia coli strains and to develop infectious progeny from such lysogenized strains. The stx(2) gene of the patient E. coli O157:H7 isolate 3538/95 was replaced by the chloramphenicol acetyltransferase (cat) gene from plasmid pACYC184. Phage phi3538(Deltastx(2)::cat) was isolated after induction of E. coli O157:H7 strain 3538/95 with mitomycin. A variety of strains of enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), Stx-producing E. coli (STEC), enterotoxigenic E. coli (ETEC), enteroaggregative E. coli (EAEC), and E. coli from the physiological stool microflora were infected with phi3538(Deltastx(2)::cat), and plaque formation and lysogenic conversion of wild-type E. coli strains were investigated. With the exception of one EIEC strain, none of the E. coli strains supported the formation of plaques when used as indicators for phi3538(Deltastx(2)::cat). However, 2 of 11 EPEC, 11 of 25 STEC, 2 of 7 EAEC, 1 of 3 EIEC, and 1 of 6 E. coli isolates from the stool microflora of healthy individuals integrated the phage in their chromosomes and expressed resistance to chloramphenicol. Following induction with mitomycin, these lysogenic strains released infectious particles of phi3538(Deltastx(2)::cat) that formed plaques on a lawn of E. coli laboratory strain C600. The results of our study demonstrate that phi3538(Deltastx(2)::cat) was able to infect and lysogenize particular enteric strains of pathogenic and nonpathogenic E. coli and that the lysogens produced infectious phage progeny. Stx-encoding bacteriophages are able to spread stx genes among enteric E. coli strains. PMID- 10473387 TI - Generalized transduction of small Yersinia enterocolitica plasmids. AB - To study phage-mediated gene transfer in Yersinia, the ability of Yersinia phages to transduce naturally occurring plasmids was investigated. The transduction experiments were performed with a temperate phage isolated from a pathogenic Yersinia enterocolitica strain and phage mixtures isolated from sewage. Small plasmids (4.3 and 5.8 kb) were transduced at a frequency of 10(-5) to 10(-7)/PFU. However, we could not detect the transduction of any indigenous virulence plasmid (ca. 72 kb) in pathogenic Yersinia strains. Transductants obtained by infection with the temperate phage were lysogenic and harbored the phage genome in their chromosomes. PMID- 10473388 TI - Enzymatic formation of G-group aflatoxins and biosynthetic relationship between G and B-group aflatoxins. AB - We detected biosynthetic activity for aflatoxins G(1) and G(2) in cell extracts of Aspergillus parasiticus NIAH-26. We found that in the presence of NADPH, aflatoxins G(1) and G(2) were produced from O-methylsterigmatocystin and dihydro O-methylsterigmatocystin, respectively. No G-group aflatoxins were produced from aflatoxin B(1), aflatoxin B(2), 5-methoxysterigmatocystin, dimethoxysterigmatocystin, or sterigmatin, confirming that B-group aflatoxins are not the precursors of G-group aflatoxins and that G- and B-group aflatoxins are independently produced from the same substrates (O-methylsterigmatocystin and dihydro-O-methylsterigmatocystin). In competition experiments in which the cell free system was used, formation of aflatoxin G(2) from dihydro-O methylsterigmatocystin was suppressed when O-methylsterigmatocystin was added to the reaction mixture, whereas aflatoxin G(1) was newly formed. This result indicates that the same enzymes can catalyze the formation of aflatoxins G(1) and G(2). Inhibition of G-group aflatoxin formation by methyrapone, SKF-525A, or imidazole indicated that a cytochrome P-450 monooxygenase may be involved in the formation of G-group aflatoxins. Both the microsome fraction and a cytosol protein with a native mass of 220 kDa were necessary for the formation of G-group aflatoxins. Due to instability of the microsome fraction, G-group aflatoxin formation was less stable than B-group aflatoxin formation. The ordA gene product, which may catalyze the formation of B-group aflatoxins, also may be required for G-group aflatoxin biosynthesis. We concluded that at least three reactions, catalyzed by the ordA gene product, an unstable microsome enzyme, and a 220-kDa cytosol protein, are involved in the enzymatic formation of G-group aflatoxins from either O-methylsterigmatocystin or dihydro-O methylsterigmatocystin. PMID- 10473389 TI - Identification of an NADH-cytochrome b(5) reductase gene from an arachidonic acid producing fungus, Mortierella alpina 1S-4, by sequencing of the encoding cDNA and heterologous expression in a fungus, Aspergillus oryzae. AB - Based on the sequence information for bovine and yeast NADH-cytochrome b(5) reductases (CbRs), a DNA fragment was cloned from Mortierella alpina 1S-4 after PCR amplification. This fragment was used as a probe to isolate a cDNA clone with an open reading frame encoding 298 amino acid residues which show marked sequence similarity to CbRs from other sources, such as yeast (Saccharomyces cerevisiae), bovine, human, and rat CbRs. These results suggested that this cDNA is a CbR gene. The results of a structural comparison of the flavin-binding beta-barrel domains of CbRs from various species and that of the M. alpina enzyme suggested that the overall barrel-folding patterns are similar to each other and that a specific arrangement of three highly conserved amino acid residues (i.e., arginine, tyrosine, and serine) plays a role in binding with the flavin (another prosthetic group) through hydrogen bonds. The corresponding genomic gene, which was also cloned from M. alpina 1S-4 by means of a hybridization method with the above probe, had four introns of different sizes. These introns had GT at the 5' end and AG at the 3' end, according to a general GT-AG rule. The expression of the full-length cDNA in a filamentous fungus, Aspergillus oryzae, resulted in an increase (4.7 times) in ferricyanide reduction activity involving the use of NADH as an electron donor in the microsomes. The M. alpina CbR was purified by solubilization of microsomes with cholic acid sodium salt, followed by DEAE Sephacel, Mono-Q HR 5/5, and AMP-Sepharose 4B affinity column chromatographies; there was a 645-fold increase in the NADH-ferricyanide reductase specific activity. The purified CbR preferred NADH over NADPH as an electron donor. This is the first report of an analysis of this enzyme in filamentous fungi. PMID- 10473390 TI - Purification of mutacin III from group III Streptococcus mutans UA787 and genetic analyses of mutacin III biosynthesis genes. AB - Previously, members of our group reported the isolation and characterization of mutacin II from Streptococcus mutans T8 and the genetic analyses of the mutacin II biosynthesis genes (J. Novak, P. W. Caufield, and E. J. Miller, J. Bacteriol. 176:4316-4320, 1994; F. Qi, P. Chen, and P. W. Caufield, Appl. Environ. Microbiol. 65:652-658, 1999; P. Chen, F. Qi, J. Novak, and P. W. Caufield, Appl. Environ. Microbiol. 65:1356-1360, 1999). In this study, we cloned and sequenced the mutacin III biosynthesis gene locus from a group III strain of S. mutans, UA787. DNA sequence analysis revealed eight open reading frames, which we designated mutR, -A, -A', -B, -C, -D, -P, and -T. MutR bears strong homology with MutR of mutacin II, while MutA, -B, -C, -D, -P, and -T are counterparts of proteins in the lantibiotic epidermin group. MutA' has 60% amino acid identity with MutA and therefore appears to be a duplicate of MutA. Insertional inactivation demonstrated that mutA is an essential gene for mutacin III production, while mutA' is not required. Mutacin III was purified to homogeneity by using reverse-phase high-pressure liquid chromatography. N-terminal peptide sequencing of the purified mutacin III determined mutA to be the structural gene for prepromutacin III. The molecular mass of the purified peptide was measured by laser disorption mass spectrophotometry and found to be 2,266.43 Da, consistent with our supposition that mutacin III has posttranslational modifications similar to those of the lantibiotic epidermin. PMID- 10473391 TI - Analysis of the gene family encoding lipases in Candida rugosa by competitive reverse transcription-PCR. AB - Synthesis of multiple extracellular lipases in Candida rugosa has been demonstrated. However, it is difficult to characterize the expression spectrum of lip genes, since the sequences of the lip multigene family are very closely related. A competitive reverse transcription-PCR assay was developed to quantify the expression of lip genes. In agreement with the protein profile, the abundance of lip mRNAs was found to be (in decreasing order) lip1, lip3, lip2, lip5, and lip4. To analyze the effects of different culture conditions, the transcript concentrations for these mRNA species were normalized relative to the values for gpd, encoding glyceraldehyde-3-phosphate dehydrogenase. In relative terms, lip1 and lip3 were highly and constitutively expressed (about 10(5) molecules per microg of total RNA) whereas the other inducible lip genes, especially lip4, showed significant changes in mRNA expression under different culture conditions. These results indicate that differential transcriptional control of lip genes results in multiple forms of lipase proteins. PMID- 10473392 TI - Occurrence of Shewanella algae in Danish coastal water and effects of water temperature and culture conditions on its survival. AB - The marine bacterium Shewanella algae, which was identified as the cause of human cases of bacteremia and ear infections in Denmark in the summers of 1994 and 1995, was detected in seawater only during the months (July, August, September, and October) when the water temperature was above 13 degrees C. The bacterium is a typical mesophilic organism, and model experiments were conducted to elucidate the fate of the organism under cold and nutrient-limited conditions. The culturable count of S. algae decreased rapidly from 10(7) CFU/ml to 10(1) CFU/ml in approximately 1 month when cells grown at 20 to 37 degrees C were exposed to cold (2 degrees C) seawater. In contrast, the culturable count of cells exposed to warmer seawater (10 to 25 degrees C) remained constant. Allowing the bacterium a transition period in seawater at 20 degrees C before exposure to the 2 degrees C seawater resulted in 100% survival over a period of 1 to 2 months. The cold protection offered by this transition (starvation) probably explains the ability of the organism to persist in Danish seawater despite very low (0 to 1 degrees C) winter water temperatures. The culturable counts of samples kept at 2 degrees C increased to 10(5) to 10(7) CFU/ml at room temperature. Most probable number analysis showed this result to be due to regrowth rather than resuscitation. It was hypothesized that S. algae would survive cold exposure better if in the biofilm state; however, culturable counts from S. algae biofilms decreased as rapidly as did counts of planktonic cells. PMID- 10473393 TI - Construction of environmental DNA libraries in Escherichia coli and screening for the presence of genes conferring utilization of 4-hydroxybutyrate. AB - Environmental DNA libraries from three different soil samples were constructed. The average insert size was 5 to 8 kb and the percentage of plasmids with inserts was approximately 80%. The recombinant Escherichia coli strains (approximately 930,000) were screened for 4-hydroxybutyrate utilization. Thirty-six positive E. coli clones were obtained during the initial screen, and five of them contained a recombinant plasmid (pAH1 to pAH5) which conferred a stable 4-hydroxybutyrate positive phenotype. These E. coli clones were studied further. All five were able to grow with 4-hydroxybutyrate as sole carbon and energy source and exhibited 4 hydroxybutyrate dehydrogenase activity in crude extracts. Sequencing of pAH5 revealed a gene homologous to the gbd gene of Ralstonia eutropha, which encodes a 4-hydroxybutyrate dehydrogenase. Two other genes (orf1 and orf6) conferring utilization of 4-hydroxybutyrate were identified during subcloning and sequencing of the inserts of pAH1 and pAH3. The deduced orf1 gene product showed similarities to members of the DedA family of proteins. The sequence of the deduced orf6 gene product harbors the fingerprint pattern of enoyl-coenzyme A hydratases/isomerases. The other sequenced inserts of the plasmids recovered from the positive clones revealed no significant similarity to any other gene or gene product whose sequence is available in the National Center for Biotechnology Information databases. PMID- 10473394 TI - Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed field gel electrophoresis in typing of Lactobacillus rhamnosus and L. casei strains. AB - A total of 24 strains, biochemically identified as members of the Lactobacillus casei group, were identified by PCR with species-specific primers. The same set of strains was typed by randomly amplified polymorphic DNA (RAPD) analysis, ribotyping, and pulsed-field gel electrophoresis (PFGE) in order to compare the discriminatory power of the methods. Species-specific primers for L. rhamnosus and L. casei identified the type strain L. rhamnosus ATCC 7469 and the neotype strain L. casei ATCC 334, respectively, but did not give any signal with the recently revived species L. zeae, which contains the type strain ATCC 15820 and the strain ATCC 393, which was previously classified as L. casei. Our results are in accordance with the suggested new classification of the L. casei group. Altogether, 21 of the 24 strains studied were identified with the species specific primers. In strain typing, PFGE was the most discriminatory method, revealing 17 genotypes for the 24 strains studied. Ribotyping and RAPD analysis yielded 15 and 12 genotypes, respectively. PMID- 10473395 TI - Inactivation of MXR1 abolishes formation of dimethyl sulfide from dimethyl sulfoxide in Saccharomyces cerevisiae. AB - Dimethyl sulfide (DMS) is a sulfur compound of importance for the organoleptic properties of beer, especially some lager beers. Synthesis of DMS during beer production occurs partly during wort production and partly during fermentation. Methionine sulfoxide reductases are the enzymes responsible for reduction of oxidized cellular methionines. These enzymes have been suggested to be able to reduce dimethyl sulfoxide (DMSO) as well, with DMS as the product. A gene for an enzymatic activity leading to methionine sulfoxide reduction in Saccharomyces yeast was recently identified. We confirmed that the Saccharomyces cerevisiae open reading frame YER042w appears to encode a methionine sulfoxide reductase, and propose the name MXR1 for the gene. We found that Mxr1p catalyzes reduction of DMSO to DMS and that an mxr1 disruption mutant cannot reduce DMSO to DMS. Mutant strains appear to have unchanged fitness under several laboratory conditions, and in this paper I hypothesize that disruption of MXR1 in brewing yeasts would neutralize the contribution of the yeast to the DMS content in beer. PMID- 10473396 TI - Glucose metabolism and kinetics of phosphorus removal by the fermentative bacterium Microlunatus phosphovorus. AB - Phosphorus and carbon metabolism in Microlunatus phosphovorus was investigated by using a batch reactor to study the kinetics of uptake and release of extracellular compounds, in combination with (31)P and (13)C nuclear magnetic resonance (NMR) to characterize intracellular pools and to trace the fate of carbon substrates through the anaerobic and aerobic cycles. The organism was subjected to repetitive anaerobic and aerobic cycles to induce phosphorus release and uptake in a sequential batch reactor; an ultrafiltration membrane module was required since cell suspensions did not sediment. M. phosphovorus fermented glucose to acetate via an Embden-Meyerhof pathway but was unable to grow under anaerobic conditions. A remarkable time shift was observed between the uptake of glucose and excretion of acetate, resulting in an intracellular accumulation of acetate. The acetate produced was oxidized in the subsequent aerobic stage. Very high phosphorus release and uptake rates were measured, 3.34 mmol g of cell(-1) h(-1) and 1.56 mmol g of cell(-1) h(-1), respectively, values only comparable with those determined in activated sludge. In the aerobic period, growth was strictly dependent on the availability of external phosphate. Natural abundance (13)C NMR showed the presence of reserves of glutamate and trehalose in cell suspensions. Unexpectedly, [1-(13)C]glucose was not significantly channeled to the synthesis of internal reserves in the anaerobic phase, and acetate was not during the aerobic stage, although the glutamate pool became labeled via the exchange with intermediates of the tricarboxylic acid cycle at the level of glutamate dehydrogenase. The intracellular pool of glutamate increased under anaerobic conditions and decreased during the aerobic period. The contribution of M. phosphovorus for phosphorus removal in wastewater treatment plants is discussed on the basis of the metabolic features disclosed by this study. PMID- 10473398 TI - A most-probable-number assay for enumeration of infectious Cryptosporidium parvum oocysts. AB - Cryptosporidium is globally established as a contaminant of drinking and recreational waters. A previously described cell culture infectivity assay capable of detecting infectious oocysts was adapted to quantify viable oocysts through sporozoite invasion and clustering of foci. Eight experiments were performed by using oocysts less than 4 months of age to inoculate host HCT-8 cell monolayers. Oocysts were diluted in a standard 5- or 10-fold multiple dilution format, levels of infection and clustering were determined, and the most probable number (MPN) of infectious oocysts in the stock suspension was calculated. The MPN was compared to the initial oocyst inoculum to determine the level of correlation. For oocysts less than 30 days of age, the correlation coefficient (r) was 0.9726 (0.9306 to 0.9893; n = 20). A two-tailed P value (alpha = 0.05) indicated that P was less than 0.0001. This strong correlation suggests that the MPN can be used to effectively enumerate infectious oocysts in a cell culture system. Age affected the degree of oocyst infectivity. Oocyst infectivity was tested by the focus detection method (FDM)-MPN assay and in BALB/c mice before and after treatment with pulsed white light (PureBrite). The FDM-MPN assay and animal infectivity assays both demonstrated more than a 4 log(10) inactivation. Municipal water systems and a host of other water testing organizations could utilize the FDM-MPN assay for routine survival and disinfection studies. PMID- 10473397 TI - Purification and characterization of the soluble methane monooxygenase of the type II methanotrophic bacterium Methylocystis sp. strain WI 14. AB - Methane monooxygenase (MMO) catalyzes the oxidation of methane to methanol as the first step of methane degradation. A soluble NAD(P)H-dependent methane monooxygenase (sMMO) from the type II methanotrophic bacterium WI 14 was purified to homogeneity. Sequencing of the 16S rDNA and comparison with that of other known methanotrophic bacteria confirmed that strain WI 14 is very close to the genus Methylocystis. The sMMO is expressed only during growth under copper limitation (<0.1 microM) and with ammonium or nitrate ions as the nitrogen source. The enzyme exhibits a low substrate specificity and is able to oxidize several alkanes and alkenes, cyclic hydrocarbons, aromatics, and halogenic aromatics. It has three components, hydroxylase, reductase and protein B, which is involved in enzyme regulation and increases sMMO activity about 10-fold. The relative molecular masses of the native components were estimated to be 229, 41, and 18 kDa, respectively. The hydroxylase contains three subunits with relative molecular masses of 57, 43, and 23 kDa, which are present in stoichiometric amounts, suggesting that the native protein has an alpha(2)beta(2)gamma(2) structure. We detected 3.6 mol of iron per mol of hydroxylase by atomic absorption spectrometry. sMMO is strongly inhibited by Hg(2+) ions (with a total loss of enzyme activity at 0.01 mM Hg(2+)) and Cu(2+), Zn(2+), and Ni(2+) ions (95, 80, and 40% loss of activity at 1 mM ions). The complete sMMO gene sequence has been determined. sMMO genes from strain WI 14 are clustered on the chromosome and show a high degree of homology (at both the nucleotide and amino acid levels) to the corresponding genes from Methylosinus trichosporium OB3b, Methylocystis sp. strain M, and Methylococcus capsulatus (Bath). PMID- 10473399 TI - Long-chain polyphosphate causes cell lysis and inhibits Bacillus cereus septum formation, which is dependent on divalent cations. AB - We investigated the cellular mechanisms that led to growth inhibition, morphological changes, and lysis of Bacillus cereus WSBC 10030 when it was challenged with a long-chain polyphosphate (polyP). At a concentration of 0.1% or higher, polyP had a bacteriocidal effect on log-phase cells, in which it induced rapid lysis and reductions in viable cell counts of up to 3 log units. The cellular debris consisted of empty cell wall cylinders and polar caps, suggesting that polyP-induced lysis was spatially specific. This activity was strictly dependent on active growth and cell division, since polyP failed to induce lysis in cells treated with chloramphenicol and in stationary-phase cells, which were, however, bacteriostatically inhibited by polyP. Similar observations were made with B. cereus spores; 0.1% polyP inhibited spore germination and outgrowth, and a higher concentration (1.0%) was even sporocidal. Supplemental divalent metal ions (Mg(2+) and Ca(2+)) could almost completely block and reverse the antimicrobial activity of polyP; i. e., they could immediately stop lysis and reinitiate rapid cell division and multiplication. Interestingly, a sublethal polyP concentration (0.05%) led to the formation of elongated cells (average length, 70 microm) after 4 h of incubation. While DNA replication and chromosome segregation were undisturbed, electron microscopy revealed a complete lack of septum formation within the filaments. Exposure to divalent cations resulted in instantaneous formation and growth of ring-shaped edges of invaginating septal walls. After approximately 30 min, septation was complete, and cell division resumed. We frequently observed a minicell-like phenotype and other septation defects, which were probably due to hyperdivision activity after cation supplementation. We propose that polyP may have an effect on the ubiquitous bacterial cell division protein FtsZ, whose GTPase activity is known to be strictly dependent on divalent metal ions. It is tempting to speculate that polyP, because of its metal ion-chelating nature, indirectly blocks the dynamic formation (polymerization) of the Z ring, which would explain the aseptate phenotype. PMID- 10473400 TI - Nested allele-specific PCR primers distinguish genetic groups of Uncinula necator. AB - Isolates of the obligately biotrophic fungus Uncinula necator cluster in three distinct genetic groups (groups I, II, and III). We designed PCR primers specific for these groups in order to monitor field populations of U. necator. We used the nucleotide sequences of the gene that encodes eburicol 14alpha-demethylase (CYP51) and of the ribosomal DNA internal transcribed spacer 1 (ITS1), ITS2, and 5. 8S regions. We identified four point mutations (three in CYP51 and one in ITS1) that distinguished groups I and II from group III based on a sample of 132 single-spore isolates originating from Europe, Tunisia, Israel, India, and Australia. We developed a nested allele-specific PCR assay in which the CYP51 point mutations were used to detect and distinguish groups I and II from group III in crude mildewed samples from vineyards. In a preliminary study performed with samples from French vineyards in which isolates belonging to genetic groups I and III were present, we found that a shift from a population composed primarily of group I isolates to a population composed primarily of group III isolates occurred during the grapevine growing season. PMID- 10473402 TI - An alpha-L-arabinofuranosidase from Trichoderma reesei containing a noncatalytic xylan-binding domain. AB - L-Sorbose, an excellent cellulase and xylanase inducer from Trichoderma reesei PC 3-7, also induced alpha-L-arabinofuranosidase (alpha-AF) activity. An alpha-AF induced by L-sorbose was purified to homogeneity, and its molecular mass was revealed to be 35 kDa (AF35), which was not consistent with that of the previously reported alpha-AF. Another species, with a molecular mass of 53 kDa (AF53), which is identical to that of the reported alpha-AF, was obtained by a different purification procedure. Acid treatment of the ammonium sulfate precipitated fraction at pH 3.0 in the purification steps or pepsin treatment of the purified AF53 reduced the molecular mass to 35 kDa. Both purified enzymes have the same enzymological properties, such as pH and temperature effects on activity and kinetic parameters for p-nitrophenyl-alpha-L-arabinofuranoside (pNPA). Moreover, the N-terminal amino acid sequences of these enzymes were identical with that of the reported alpha-AF. Therefore, it is obvious that AF35 results from the proteolytic cleavage of the C-terminal region of AF53. Although AF35 and AF53 showed the same catalytic constant with pNPA, the former showed drastically reduced specific activity against oat spelt xylan compared to the latter. Furthermore, AF53 was bound to xylan rather than to crystalline cellulose (Avicel), but AF35 could not be bound to any of the glycans. These results suggest that AF53 is a modular glycanase, which consists of an N-terminal catalytic domain and a C-terminal noncatalytic xylan-binding domain. PMID- 10473401 TI - Cloning of the gene encoding a novel thermostable alpha-galactosidase from Thermus brockianus ITI360. AB - An alpha-galactosidase gene from Thermus brockianus ITI360 was cloned, sequenced, and expressed in Escherichia coli, and the recombinant protein was purified. The gene, designated agaT, codes for a 476-residue polypeptide with a calculated molecular mass of 53, 810 Da. The native structure of the recombinant enzyme (AgaT) was estimated to be a tetramer. AgaT displays amino acid sequence similarity to the alpha-galactosidases of Thermotoga neapolitana and Thermotoga maritima and a low-level sequence similarity to alpha-galactosidases of family 36 in the classification of glycosyl hydrolases. The enzyme is thermostable, with a temperature optimum of activity at 93 degrees C with para-nitrophenyl-alpha galactopyranoside as a substrate. Half-lives of inactivation at 92 and 80 degrees C are 100 min and 17 h, respectively. The pH optimum is between 5.5 and 6.5. The enzyme displayed high affinity for oligomeric substrates. The K(m)s for melibiose and raffinose at 80 degrees C were determined as 4.1 and 11.0 mM, respectively. The alpha-galactosidase gene in T. brockianus ITI360 was inactivated by integrational mutagenesis. Consequently, no alpha-galactosidase activity was detectable in crude extracts of the mutant strain, and it was unable to use melibiose or raffinose as a single carbohydrate source. PMID- 10473403 TI - Purification and characterization of an extracellular protease from the fish pathogen Yersinia ruckeri and effect of culture conditions on production. AB - A novel protease, hydrolyzing azocasein, was identified, purified, and characterized from the culture supernatant of the fish pathogen Yersinia ruckeri. Exoprotease production was detected at the end of the exponential growth phase and was temperature dependent. Activity was detected in peptone but not in Casamino Acid medium. Its synthesis appeared to be under catabolite repression and ammonium control. The protease was purified in a simple two-step procedure involving ammonium sulfate precipitation and ion-exchange chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the purified protein indicated an estimated molecular mass of 47 kDa. The protease had characteristics of a cold-adapted protein, i.e., it was more active in the range of 25 to 42 degrees C and had an optimum activity at 37 degrees C. The activation energy for the hydrolysis of azocasein was determined to be 15.53 kcal/mol, and the enzyme showed a rapid decrease in activity at 42 degrees C. The enzyme had an optimum pH of around 8. Characterization of the protease showed that it required certain cations such as Mg(2+) or Ca(2+) for maximal activity and was inhibited by EDTA, 1,10-phenanthroline, and EGTA but not by phenylmethylsulfonyl fluoride. Two N-methyl-N-nitro-N-nitrosoguanidine mutants were isolated and analyzed; one did not show caseinolytic activity and lacked the 47-kDa protein, while the other was hyperproteolytic and produced increased amounts of the 47-kDa protein. Azocasein activity, SDS-PAGE, immunoblotting by using polyclonal anti-47-kDa-protease serum, and zymogram analyses showed that protease activity was present in 8 of 14 strains tested and that two Y. ruckeri groups could be established based on the presence or absence of the 47-kDa protease. PMID- 10473404 TI - Phylogenetic affiliation and quantification of psychrophilic sulfate-reducing isolates in marine Arctic sediments. AB - Thirteen psychrophilic sulfate-reducing isolates from two permanently cold fjords of the Arctic island Spitsbergen (Hornsund and Storfjord) were phylogenetically analyzed. They all belonged to the delta subclass of Proteobacteria and were widely distributed within this group, indicating that psychrophily is a polyphyletic property. A new 16S rRNA-directed oligonucleotide probe was designed against the largest coherent cluster of these isolates. The new probe, as well as a set of available probes, was applied in rRNA slot blot hybridization to investigate the composition of the sulfate-reducing bacterial community in the sediments. rRNA related to the new cluster of incompletely oxidizing, psychrophilic isolates made up 1.4 to 20.9% of eubacterial rRNA at Storfjord and 0.6 to 3. 5% of eubacterial rRNA at Hornsund. This group was the second-most abundant group of sulfate reducers at these sites. Denaturing gradient gel electrophoresis and hybridization analysis showed bands identical to those produced by our isolates. The data indicate that the psychrophilic isolates are quantitatively important in Svalbard sediments. PMID- 10473405 TI - High bacterial diversity in permanently cold marine sediments. AB - A 16S ribosomal DNA (rDNA) clone library from permanently cold marine sediments was established. Screening 353 clones by dot blot hybridization with group specific oligonucleotide probes suggested a predominance of sequences related to bacteria of the sulfur cycle (43.4% potential sulfate reducers). Within this fraction, the major cluster (19.0%) was affiliated with Desulfotalea sp. and other closely related psychrophilic sulfate reducers isolated from the same habitat. The cloned sequences showed between 93 and 100% similarity to these bacteria. Two additional groups were frequently encountered: 13% of the clones were related to Desulfuromonas palmitatis, and a second group was affiliated with Myxobacteria spp. and Bdellovibrio spp. Many clones (18.1%) belonged to the gamma subclass of the class Proteobacteria and were closest to symbiotic or free-living sulfur oxidizers. Probe target groups were further characterized by amplified rDNA restriction analysis to determine diversity within the groups and within the clone library. Rarefaction analysis suggested that the total diversity assessed by 16S rDNA analysis was very high in these permanently cold sediments and was only partially revealed by screening of 353 clones. PMID- 10473406 TI - Characterization of an acetyl xylan esterase from the anaerobic fungus Orpinomyces sp. strain PC-2. AB - A 1,067-bp cDNA, designated axeA, coding for an acetyl xylan esterase (AxeA) was cloned from the anaerobic rumen fungus Orpinomyces sp. strain PC-2. The gene had an open reading frame of 939 bp encoding a polypeptide of 313 amino acid residues with a calculated mass of 34,845 Da. An active esterase using the original start codon of the cDNA was synthesized in Escherichia coli. Two active forms of the esterase were purified from recombinant E. coli cultures. The size difference of 8 amino acids was a result of cleavages at two different sites within the signal peptide. The enzyme released acetate from several acetylated substrates, including acetylated xylan. The activity toward acetylated xylan was tripled in the presence of recombinant xylanase A from the same fungus. Using p-nitrophenyl acetate as a substrate, the enzyme had a K(m) of 0.9 mM and a V(max) of 785 micromol min(-1) mg(-1). It had temperature and pH optima of 30 degrees C and 9.0, respectively. AxeA had 56% amino acid identity with BnaA, an acetyl xylan esterase of Neocallimastix patriciarum, but the Orpinomyces AxeA was devoid of a noncatalytic repeated peptide domain (NCRPD) found at the carboxy terminus of the Neocallimastix BnaA. The NCRPD found in many glycosyl hydrolases and esterases of anaerobic fungi has been postulated to function as a docking domain for cellulase hemicellulase complexes, similar to the dockerin of the cellulosome of Clostridium thermocellum. The difference in domain structures indicated that the two highly similar esterases of Orpinomyces and Neocallimastix may be differently located, the former being a free enzyme and the latter being a component of a cellulase-hemicellulase complex. Sequence data indicate that AxeA and BnaA might represent a new family of hydrolases. PMID- 10473407 TI - Comparison of bacterial community structures in the rhizoplane of tomato plants grown in soils suppressive and conducive towards bacterial wilt. AB - It has been reported that the growth of Ralstonia solanacearum is suppressed at the rhizoplane of tomato plants and that tomato bacterial wilt is suppressed in plants grown in a soil (Mutsumi) in Japan. To evaluate the biological factors contributing to the suppressiveness of the soil in three treated Mutsumi soils (chloroform fumigated soil; autoclaved soil mixed with intact Mutsumi soil; and autoclaved soil mixed with intact, wilt-conducive Yamadai soil) infested with R. solanacearum, we bioassayed soil samples for tomato bacterial wilt. Chloroform fumigation increased the extent of wilt disease. More of the tomato plant samples wilted when mixed with Yamadai soil than when mixed with Mutsumi soil. Consequently, the results indicate that the naturally existing population of microorganisms in Mutsumi soil was significantly able to reduce the severity of bacterial wilt of tomato plants. To characterize the types of bacteria present at the rhizoplane, we isolated rhizoplane bacteria and classified them into 22 groups by comparing their 16S restriction fragment length polymorphism patterns. In Yamadai soil a single group of bacteria was extremely predominant (73.1%), whereas in Mutsumi soil the distribution of the bacterial groups was much more even. The 16S rDNA sequence analysis of strains of dominant groups suggested that gram-negative bacteria close to the beta-proteobacteria were most common at the rhizoplane of the tomato plants. During in vitro assays, rhizoplane bacteria in Mutsumi soil grew more vigorously on pectin, one of the main root exudates of tomato, compared with those in Yamadai soil. Our results imply that it is difficult for the pathogen to dominate in a diversified rhizobacterial community that thrives on pectin. PMID- 10473408 TI - D-Lactate dehydrogenase gene (ldhD) inactivation and resulting metabolic effects in the Lactobacillus johnsonii strains La1 and N312. AB - Lactobacillus johnsonii La1, a probiotic bacterium with demonstrated health effects, grows in milk, where it ferments lactose to D- and L-lactate in a 60:40% ratio. The D-lactate dehydrogenase (D-LDH) gene (ldhD) of this strain was isolated, and an in vitro-truncated copy of that gene was used to inactivate the genomic copy in two strains, La1 and N312, by gene replacement. For that, an 8-bp deletion was generated within the cloned ldhD gene to inactivate its function. The plasmid containing the altered ldhD was transferred to L. johnsonii via conjugative comobilization with Lactococcus lactis carrying pAMbeta1. Crossover integrations of the plasmid at the genomic ldhD site were selected, and appropriate resolution of the cointegrate structures resulted in mutants that had lost the plasmid and in which the original ldhD was replaced by the truncated copy. These mutants completely lacked D-LDH activity. Nevertheless, the lower remaining L-LDH activity of the cells was sufficient to reroute most of the accumulating pyruvate to L-lactate. Only a marginal increase in production of the secondary end products acetaldehyde, diacetyl, and acetoin was observed. It can be concluded that in L. johnsonii D- and L-LDH are present in substantial excess for their role to eliminate pyruvate and regenerate NAD(+) and that accumulated pyruvate is therefore not easily redirected in high amounts to secondary metabolic routes. PMID- 10473409 TI - Nitrification and autotrophic nitrifying bacteria in a hydrocarbon-polluted soil. AB - In vitro ammonia-oxidizing bacteria are capable of oxidizing hydrocarbons incompletely. This transformation is accompanied by competitive inhibition of ammonia monooxygenase, the first key enzyme in nitrification. The effect of hydrocarbon pollution on soil nitrification was examined in situ. In a microcosm study, adding diesel fuel hydrocarbon to an uncontaminated soil (agricultural unfertilized soil) treated with ammonium sulfate dramatically reduced the amount of KCl-extractable nitrate but stimulated ammonium consumption. In a soil with long history of pollution that was treated with ammonium sulfate, 90% of the ammonium was transformed into nitrate after 3 weeks of incubation. Nitrate production was twofold higher in the contaminated soil than in the agricultural soil to which hydrocarbon was not added. To assess if ammonia-oxidizing bacteria acquired resistance to inhibition by hydrocarbon, the contaminated soil was reexposed to diesel fuel. Ammonium consumption was not affected, but nitrate production was 30% lower than nitrate production in the absence of hydrocarbon. The apparent reduction in nitrification resulted from immobilization of ammonium by hydrocarbon-stimulated microbial activity. These results indicated that the hydrocarbon inhibited nitrification in the noncontaminated soil (agricultural soil) and that ammonia-oxidizing bacteria in the polluted soil acquired resistance to inhibition by the hydrocarbon, possibly by increasing the affinity of nitrifying bacteria for ammonium in the soil. PMID- 10473410 TI - Cold-adapted alanine dehydrogenases from two antarctic bacterial strains: gene cloning, protein characterization, and comparison with mesophilic and thermophilic counterparts. AB - The genes encoding NAD(+)-dependent alanine dehydrogenases (AlaDHs) (EC 1.4.1.1) from the Antarctic bacterial organisms Shewanella sp. strain Ac10 (SheAlaDH) and Carnobacterium sp. strain St2 (CarAlaDH) were cloned and expressed in Escherichia coli. Of all of the AlaDHs that have been sequenced, SheAlaDH exhibited the highest level of sequence similarity to the AlaDH from the gram-negative bacterium Vibrio proteolyticus (VprAlaDH). CarAlaDH was most similar to AlaDHs from mesophilic and thermophilic Bacillus strains. SheAlaDH and CarAlaDH had features typical of cold-adapted enzymes; both the optimal temperature for catalytic activity and the temperature limit for retaining thermostability were lower than the values obtained for the mesophilic counterparts. The k(cat)/K(m) value for the SheAlaDH reaction was about three times higher than the k(cat)/K(m) value for VprAlaDH, but it was much lower than the k(cat)/K(m) value for the AlaDH from Bacillus subtilis. Homology-based structural models of various AlaDHs, including the two psychotropic AlaDHs, were constructed. The thermal instability of SheAlaDH and CarAlaDH may result from relatively low numbers of salt bridges in these proteins. PMID- 10473411 TI - Degradation of 4-fluorobiphenyl by mycorrhizal fungi as determined by (19)F nuclear magnetic resonance spectroscopy and (14)C radiolabelling analysis. AB - The pathways of biotransformation of 4-fluorobiphenyl (4FBP) by the ectomycorrhizal fungus Tylospora fibrilosa and several other mycorrhizal fungi were investigated by using (19)F nuclear magnetic resonance (NMR) spectroscopy in combination with (14)C radioisotope-detected high-performance liquid chromatography ((14)C-HPLC). Under the conditions used in this study T. fibrillosa and some other species degraded 4FBP. (14)C-HPLC profiles indicated that there were four major biotransformation products, whereas (19)F NMR showed that there were six major fluorine-containing products. We confirmed that 4 fluorobiphen-4'-ol and 4-fluorobiphen-3'-ol were two of the major products formed, but no other products were conclusively identified. There was no evidence for the expected biotransformation pathway (namely, meta cleavage of the less halogenated ring), as none of the expected products of this route were found. To the best of our knowledge, this is the first report describing intermediates formed during mycorrhizal degradation of halogenated biphenyls. PMID- 10473412 TI - Microbial proline 4-hydroxylase screening and gene cloning. AB - Microbial proline 4-hydroxylases, which hydroxylate free L-proline to trans-4 hydroxy-L-proline, were screened in order to establish an industrial system for biotransformation of L-proline to trans-4-hydroxy-L-proline. Enzyme activities were detected in eight strains, including strains of Dactylosporangium spp. and Amycolatopsis spp. The Dactylosporangium sp. strain RH1 enzyme was partially purified 3,300-fold and was estimated to be a monomer polypeptide with an apparent molecular mass of 31 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Degenerate primers based on the N-terminal amino acid sequence of the 31-kDa polypeptide were synthesized in order to amplify the corresponding 71-bp DNA fragment. A 5.5-kbp DNA fragment was isolated by using the 71-bp fragment labeled with digoxigenin as a probe for a genomic library of Dactylosporangium sp. strain RH1 constructed in Escherichia coli. One of the open reading frames found in the cloned DNA, which encoded a 272-amino-acid polypeptide (molecular mass, 29, 715 daltons), was thought to be a proline 4 hydroxylase gene. The gene was expressed in E. coli as a fused protein with the N terminal 34 amino acids of the beta-galactosidase alpha-fragment. The E. coli recombinant exhibited proline 4-hydroxylase activity that was 13. 6-fold higher than the activity in the original strain, Dactylosporangium sp. strain RH1. No homology was detected with other 2-oxoglutarate-dependent dioxygenases when databases were searched; however, the histidine motif conserved in 2-oxoglutarate dependent dioxygenases was found in the gene. PMID- 10473413 TI - Development and field performance of a broad-spectrum nonviable asporogenic recombinant strain of Bacillus thuringiensis with greater potency and UV resistance. AB - The main problems with Bacillus thuringiensis products for pest control are their often narrow activity spectrum, high sensitivity to UV degradation, and low cost effectiveness (high potency required). We constructed a sporulation-deficient SigK(-) B. thuringiensis strain that expressed a chimeric cry1C/Ab gene, the product of which had high activity against various lepidopteran pests, including Spodoptera littoralis (Egyptian cotton leaf worm) and Spodoptera exigua (lesser [beet] armyworm), which are not readily controlled by other Cry delta-endotoxins. The SigK(-) host strain carried the cry1Ac gene, the product of which is highly active against the larvae of the major pests Ostrinia nubilalis (European corn borer) and Heliothis virescens (tobacco budworm). This new strain had greater potency and a broader activity spectrum than the parent strain. The crystals produced by the asporogenic strain remained encapsulated within the cells, which protected them from UV degradation. The cry1C/Ab gene was introduced into the B. thuringiensis host via a site-specific recombination vector so that unwanted DNA was eliminated. Therefore, the final construct contained no sequences of non-B. thuringiensis origin. As the recombinant strain is a mutant blocked at late sporulation, it does not produce viable spores and therefore cannot compete with wild-type B. thuringiensis strains in the environment. It is thus a very safe biopesticide. In field trials, this new recombinant strain protected cabbage and broccoli against a pest complex under natural infestation conditions. PMID- 10473414 TI - Identification of an ATP-driven, osmoregulated glycine betaine transport system in Listeria monocytogenes. AB - The ability of the gram-positive, food-borne pathogen Listeria monocytogenes to tolerate environments of elevated osmolarity and reduced temperature is due in part to the transport and accumulation of the osmolyte glycine betaine. Previously we showed that glycine betaine transport was the result of Na(+) glycine betaine symport. In this report, we identify a second glycine betaine transporter from L. monocytogenes which is osmotically activated but does not require a high concentration of Na(+) for activity. By using a pool of Tn917-LTV3 mutants, a salt- and chill-sensitive mutant which was also found to be impaired in its ability to transport glycine betaine was isolated. DNA sequence analysis of the region flanking the site of transposon insertion revealed three open reading frames homologous to opuA from Bacillus subtilis and proU from Escherichia coli, both of which encode glycine betaine transport systems that belong to the superfamily of ATP-dependent transporters. The three open reading frames are closely spaced, suggesting that they are arranged in an operon. Moreover, a region upstream from the first reading frame was found to be homologous to the promoter regions of both opuA and proU. One unusual feature not shared with these other two systems is that the start codons for two of the open reading frames in L. monocytogenes appear to be TTG. That glycine betaine uptake is nearly eliminated in the mutant strain when it is assayed in the absence of Na(+) is an indication that only the ATP-dependent transporter and the Na(+) glycine betaine symporter occur in L. monocytogenes. PMID- 10473415 TI - Fraction of electrons consumed in electron acceptor reduction and hydrogen thresholds as indicators of halorespiratory physiology. AB - Measurements of the hydrogen consumption threshold and the tracking of electrons transferred to the chlorinated electron acceptor (f(e)) reliably detected chlororespiratory physiology in both mixed cultures and pure cultures capable of using tetrachloroethene, cis-1, 2-dichloroethene, vinyl chloride, 2-chlorophenol, 3-chlorobenzoate, 3-chloro-4-hydroxybenzoate, or 1,2-dichloropropane as an electron acceptor. Hydrogen was consumed to significantly lower threshold concentrations of less than 0.4 ppmv compared with the values obtained for the same cultures without a chlorinated compound as an electron acceptor. The f(e) values ranged from 0.63 to 0.7, values which are in good agreement with theoretical calculations based on the thermodynamics of reductive dechlorination as the terminal electron-accepting process. In contrast, a mixed methanogenic culture that cometabolized 3-chlorophenol exhibited a significantly lower f(e) value, 0.012. PMID- 10473416 TI - Identification of the Pseudomonas stutzeri OX1 toluene-o-xylene monooxygenase regulatory gene (touR) and of its cognate promoter. AB - Toluene-o-xylene monooxygenase is an enzymatic complex, encoded by the touABCDEF genes, responsible for the early stages of toluene and o-xylene degradation in Pseudomonas stutzeri OX1. In order to identify the loci involved in the transcriptional regulation of the tou gene cluster, deletion analysis and complementation studies were carried out with Pseudomonas putida PaW340 as a heterologous host harboring pFB1112, a plasmid that allowed regulated expression, inducible by toluene and o-xylene and their corresponding phenols, of the toluene o-xylene monooxygenase. A locus encoding a positive regulator, designated touR, was mapped downstream from the tou gene cluster. TouR was found to be similar to transcriptional activators of aromatic compound catabolic pathways belonging to the NtrC family and, in particular, to DmpR (83% similarity), which controls phenol catabolism. By using a touA-C2,3O fusion reporter system and by primer extension analysis, a TouR cognate promoter (P(ToMO)) was mapped, which showed the typical -24 TGGC, -12 TTGC sequences characteristic of sigma(54)-dependent promoters and putative upstream activating sequences. By using the reporter system described, we found that TouR responds to mono- and dimethylphenols, but not the corresponding methylbenzenes. In this respect, the regulation of the P. stutzeri system differs from that of other toluene or xylene catabolic systems, in which the hydrocarbons themselves function as effectors. Northern analyses indicated low transcription levels of tou structural genes in the absence of inducers. Basal toluene-o-xylene monooxygenase activity may thus transform these compounds to phenols, which then trigger the TouR-mediated response. PMID- 10473417 TI - Radioactive fingerprinting of microorganisms that oxidize atmospheric methane in different soils. AB - Microorganisms that oxidize atmospheric methane in soils were characterized by radioactive labelling with (14)CH(4) followed by analysis of radiolabelled phospholipid ester-linked fatty acids ((14)C-PLFAs). The radioactive fingerprinting technique was used to compare active methanotrophs in soil samples from Greenland, Denmark, the United States, and Brazil. The (14)C-PLFA fingerprints indicated that closely related methanotrophic bacteria were responsible for the oxidation of atmospheric methane in the soils. Significant amounts of labelled PLFAs produced by the unknown soil methanotrophs coeluted with a group of fatty acids that included i17:0, a17:0, and 17:1omega8c (up to 9.0% of the total (14)C-PLFAs). These PLFAs are not known to be significant constituents of methanotrophic bacteria. The major PLFAs of the soil methanotrophs (73.5 to 89.0% of the total PLFAs) coeluted with 18:1 and 18:0 fatty acids (e.g., 18:1omega9, 18:1omega7, and 18:0). The (14)C-PLFAs fingerprints of the soil methanotrophs that oxidized atmospheric methane did not change after long-term methane enrichment at 170 ppm CH(4). The (14)C-PLFA fingerprints of the soil methanotrophs were different from the PLFA profiles of type I and type II methanotrophic bacteria described previously. Some similarity at the PLFA level was observed between the unknown soil methanotrophs and the PLFA phenotype of the type II methanotrophs. Methanotrophs in Arctic, temperate, and tropical regions assimilated between 20 and 54% of the atmospheric methane that was metabolized. The lowest relative assimilation (percent) was observed for methanotrophs in agricultural soil, whereas the highest assimilation was observed for methanotrophs in rain forest soil. The results suggest that methanotrophs with relatively high carbon conversion efficiencies and very similar PLFA compositions dominate atmospheric methane metabolism in different soils. The characteristics of the methane metabolism and the (14)C-PLFA fingerprints excluded any significant role of autotrophic ammonia oxidizers in the metabolism of atmospheric methane. PMID- 10473418 TI - Molecular standardization of mating type terminology in the Gibberella fujikuroi species complex. AB - Mating type in the Gibberella fujikuroi species complex is controlled by a single locus with two alleles and is usually identified following sexual crosses with standard, female-fertile tester isolates. The mating type alleles have been arbitrarily designated "+" and "-" within each biological species, and the nomenclature is tied to the standard tester strains. We developed a pair of PCR primers that can be used to amplify a unique fragment of one of the mating type alleles (MAT-2) from at least seven of the biological species in this species complex. Based on the amplification pattern, we propose a replacement for the existing, arbitrary +/- terminology that is presently in use. The new terminology is based on DNA sequence similarities between the mating type allele fragments from the biological species of the G. fujikuroi species complex and the corresponding fragments from other filamentous ascomycetes. PMID- 10473419 TI - Identification of some of the major groups of bacteria in efficient and nonefficient biological phosphorus removal activated sludge systems. AB - To investigate the bacteria that are important to phosphorus (P) removal in activated sludge, microbial populations were analyzed during the operation of a laboratory-scale reactor with various P removal performances. The bacterial population structure, analyzed by fluorescence in situ hybridization (FISH) with oligonucleotides probes complementary to regions of the 16S and 23S rRNAs, was associated with the P removal performance of the reactor. At one stage of the reactor operation, chemical characterization revealed that extremely poor P removal was occurring. However, like in typical P-removing sludges, complete anaerobic uptake of the carbon substrate occurred. Bacteria inhibiting P removal overwhelmed the reactor, and according to FISH, bacteria of the beta subclass of the class Proteobacteria other than beta-1 or beta-2 were dominant in the sludge (58% of the population). Changes made to the operation of the reactor led to the development of a biomass population with an extremely good P removal capacity. The biochemical transformations observed in this sludge were characteristic of typical P-removing activated sludge. The microbial population analysis of the P removing sludge indicated that bacteria of the beta-2 subclass of the class Proteobacteria and actinobacteria were dominant (55 and 35%, respectively), therefore implicating bacteria from these groups in high-performance P removal. The changes in operation that led to the improved performance of the reactor included allowing the pH to rise during the anaerobic period, which promoted anaerobic phosphate release and possibly caused selection against non-phosphate removing bacteria. PMID- 10473420 TI - Oxygen-sensing reporter strain of Pseudomonas fluorescens for monitoring the distribution of low-oxygen habitats in soil. AB - The root-colonizing bacterium Pseudomonas fluorescens CHA0 was used to construct an oxygen-responsive biosensor. An anaerobically inducible promoter of Pseudomonas aeruginosa, which depends on the FNR (fumarate and nitrate reductase regulation)-like transcriptional regulator ANR (anaerobic regulation of arginine deiminase and nitrate reductase pathways), was fused to the structural lacZ gene of Escherichia coli. By inserting the reporter fusion into the chromosomal attTn7 site of P. fluorescens CHA0 by using a mini-Tn7 transposon, the reporter strain, CHA900, was obtained. Grown in glutamate-yeast extract medium in an oxystat at defined oxygen levels, the biosensor CHA900 responded to a decrease in oxygen concentration from 210 x 10(2) Pa to 2 x 10(2) Pa of O(2) by a nearly 100-fold increase in beta-galactosidase activity. Half-maximal induction of the reporter occurred at about 5 x 10(2) Pa. This dose response closely resembles that found for E. coli promoters which are activated by the FNR protein. In a carbon-free buffer or in bulk soil, the biosensor CHA900 still responded to a decrease in oxygen concentration, although here induction was about 10 times lower and the low oxygen response was gradually lost within 3 days. Introduced into a barley soil microcosm, the biosensor could report decreasing oxygen concentrations in the rhizosphere for a 6-day period. When the water content in the microcosm was raised from 60% to 85% of field capacity, expression of the reporter gene was elevated about twofold above a basal level after 2 days of incubation, suggesting that a water content of 85% caused mild anoxia. Increased compaction of the soil was shown to have a faster and more dramatic effect on the expression of the oxygen reporter than soil water content alone, indicating that factors other than the water-filled pore space influenced the oxygen status of the soil. These experiments illustrate the utility of the biosensor for detecting low oxygen concentrations in the rhizosphere and other soil habitats. PMID- 10473421 TI - Bactericidal activity of photocatalytic TiO(2) reaction: toward an understanding of its killing mechanism. AB - When titanium dioxide (TiO(2)) is irradiated with near-UV light, this semiconductor exhibits strong bactericidal activity. In this paper, we present the first evidence that the lipid peroxidation reaction is the underlying mechanism of death of Escherichia coli K-12 cells that are irradiated in the presence of the TiO(2) photocatalyst. Using production of malondialdehyde (MDA) as an index to assess cell membrane damage by lipid peroxidation, we observed that there was an exponential increase in the production of MDA, whose concentration reached 1.1 to 2.4 nmol. mg (dry weight) of cells(-1) after 30 min of illumination, and that the kinetics of this process paralleled cell death. Under these conditions, concomitant losses of 77 to 93% of the cell respiratory activity were also detected, as measured by both oxygen uptake and reduction of 2,3,5-triphenyltetrazolium chloride from succinate as the electron donor. The occurrence of lipid peroxidation and the simultaneous losses of both membrane dependent respiratory activity and cell viability depended strictly on the presence of both light and TiO(2). We concluded that TiO(2) photocatalysis promoted peroxidation of the polyunsaturated phospholipid component of the lipid membrane initially and induced major disorder in the E. coli cell membrane. Subsequently, essential functions that rely on intact cell membrane architecture, such as respiratory activity, were lost, and cell death was inevitable. PMID- 10473422 TI - Molecular and biochemical characterization of two xylanase-encoding genes from Cellulomonas pachnodae. AB - Two xylanase-encoding genes, named xyn11A and xyn10B, were isolated from a genomic library of Cellulomonas pachnodae by expression in Escherichia coli. The deduced polypeptide, Xyn11A, consists of 335 amino acids with a calculated molecular mass of 34,383 Da. Different domains could be identified in the Xyn11A protein on the basis of homology searches. Xyn11A contains a catalytic domain belonging to family 11 glycosyl hydrolases and a C-terminal xylan binding domain, which are separated from the catalytic domain by a typical linker sequence. Binding studies with native Xyn11A and a truncated derivative of Xyn11A, lacking the putative binding domain, confirmed the function of the two domains. The second xylanase, designated Xyn10B, consists of 1,183 amino acids with a calculated molecular mass of 124,136 Da. Xyn10B also appears to be a modular protein, but typical linker sequences that separate the different domains were not identified. It comprises a N-terminal signal peptide followed by a stretch of amino acids that shows homology to thermostabilizing domains. Downstream of the latter domain, a catalytic domain specific for family 10 glycosyl hydrolases was identified. A truncated derivative of Xyn10B bound tightly to Avicel, which was in accordance with the identified cellulose binding domain at the C terminus of Xyn10B on the basis of homology. C. pachnodae, a (hemi)cellulolytic bacterium that was isolated from the hindgut of herbivorous Pachnoda marginata larvae, secretes at least two xylanases in the culture fluid. Although both Xyn11A and Xyn10B had the highest homology to xylanases from Cellulomonas fimi, distinct differences in the molecular organizations of the xylanases from the two Cellulomonas species were identified. PMID- 10473423 TI - Distribution of bacterial growth activity in flow-chamber biofilms. AB - In microbial communities such as those found in biofilms, individual organisms most often display heterogeneous behavior with respect to their metabolic activity, growth status, gene expression pattern, etc. In that context, a novel reporter system for monitoring of cellular growth activity has been designed. It comprises a transposon cassette carrying fusions between the growth rate regulated Escherichia coli rrnBP1 promoter and different variant gfp genes. It is shown that the P1 promoter is regulated in the same way in E. coli and Pseudomonas putida, making it useful for monitoring of growth activity in organisms outside the group of enteric bacteria. Construction of fusions to genes encoding unstable Gfp proteins opened up the possibility of the monitoring of rates of rRNA synthesis and, in this way, allowing on-line determination of the distribution of growth activity in a complex community. With the use of these reporter tools, it is demonstrated that individual cells of a toluene-degrading P. putida strain growing in a benzyl alcohol-supplemented biofilm have different levels of growth activity which develop as the biofilm gets older. Cells that eventually grow very slowly or not at all may be stimulated to restart growth if provided with a more easily metabolizable carbon source. Thus, the dynamics of biofilm growth activity has been tracked to the level of individual cells, cell clusters, and microcolonies. PMID- 10473424 TI - Detection of viral pathogens by reverse transcriptase PCR and of microbial indicators by standard methods in the canals of the Florida Keys. AB - In order to assess the microbial water quality in canal waters throughout the Florida Keys, a survey was conducted to determine the concentration of microbial fecal indicators and the presence of human pathogenic microorganisms. A total of 19 sites, including 17 canal sites and 2 nearshore water sites, were assayed for total coliforms, fecal coliforms, Escherichia coli, Clostridium perfringens, enterococci, coliphages, F-specific (F(+)) RNA coliphages, Giardia lamblia, Cryptosporidium parvum, and human enteric viruses (polioviruses, coxsackie A and B viruses, echoviruses, hepatitis A viruses, Norwalk viruses, and small round structured viruses). Numbers of coliforms ranged from <1 to 1, 410, E. coli organisms from <1 to 130, Clostridium spp. from <1 to 520, and enterococci from <1 to 800 CFU/100 ml of sample. Two sites were positive for coliphages, but no F(+) phages were identified. The sites were ranked according to microbial water quality and compared to various water quality standards and guidelines. Seventy nine percent of the sites were positive for the presence of enteroviruses by reverse transcriptase PCR (polioviruses, coxsackie A and B viruses, and echoviruses). Sixty-three percent of the sites were positive for the presence of hepatitis A viruses. Ten percent of the sites were positive for the presence of Norwalk viruses. Ninety-five percent of the sites were positive for at least one of the virus groups. These results indicate that the canals and nearshore waters throughout the Florida Keys are being impacted by human fecal material carrying human enteric viruses through current wastewater treatment strategies such as septic tanks. Exposure to canal waters through recreation and work may be contributing to human health risks. PMID- 10473425 TI - Identification of nitrite-oxidizing bacteria with monoclonal antibodies recognizing the nitrite oxidoreductase. AB - Immunoblot analyses performed with three monoclonal antibodies (MAbs) that recognized the nitrite oxidoreductase (NOR) of the genus Nitrobacter were used for taxonomic investigations of nitrite oxidizers. We found that these MAbs were able to detect the nitrite-oxidizing systems (NOS) of the genera Nitrospira, Nitrococcus, and Nitrospina. The MAb designated Hyb 153-2, which recognized the alpha subunit of the NOR (alpha-NOR), was specific for species belonging to the genus Nitrobacter. In contrast, Hyb 153-3, which recognized the beta-NOR, reacted with nitrite oxidizers of the four genera. Hyb 153-1, which also recognized the beta-NOR, bound to members of the genera Nitrobacter and Nitrococcus. The molecular masses of the beta-NOR of the genus Nitrobacter and the beta subunit of the NOS (beta-NOS) of the genus Nitrococcus were identical (65 kDa). In contrast, the molecular masses of the beta-NOS of the genera Nitrospina and Nitrospira were different (48 and 46 kDa). When the genus-specific reactions of the MAbs were correlated with 16S rRNA sequences, they reflected the phylogenetic relationships among the nitrite oxidizers. The specific reactions of the MAbs allowed us to classify novel isolates and nitrite oxidizers in enrichment cultures at the genus level. In ecological studies the immunoblot analyses demonstrated that Nitrobacter or Nitrospira cells could be enriched from activated sludge by using various substrate concentrations. Fluorescence in situ hybridization and electron microscopic analyses confirmed these results. Permeated cells of pure cultures of members of the four genera were suitable for immunofluorescence labeling; these cells exhibited fluorescence signals that were consistent with the location of the NOS. PMID- 10473426 TI - Staphylococcal surface display of immunoglobulin A (IgA)- and IgE-specific in vitro-selected binding proteins (affibodies) based on Staphylococcus aureus protein A. AB - An expression system designed for cell surface display of hybrid proteins on Staphylococcus carnosus has been evaluated for the display of Staphylococcus aureus protein A (SpA) domains, normally binding to immunoglobulin G (IgG) Fc but here engineered by combinatorial protein chemistry to yield SpA domains, denoted affibodies, with new binding specificities. Such affibodies, with human IgA or IgE binding activity, have previously been selected from a phage library, based on an SpA domain. In this study, these affibodies have been genetically introduced in monomeric or dimeric forms into chimeric proteins expressed on the surface of S. carnosus by using translocation signals from a Staphylococcus hyicus lipase construct together with surface-anchoring regions of SpA. The recombinant surface proteins, containing the IgA- or IgE-specific affibodies, were demonstrated to be expressed as full-length proteins, localized and properly exposed at the cell surface of S. carnosus. Furthermore, these chimeric receptors were found to be functional, since recombinant S. carnosus cells were shown to have gained IgA and IgE binding capacity, respectively. In addition, a positive effect in terms of IgA and IgE reactivity was observed when dimeric versions of the affibodies were present. Potential applications for recombinant bacteria with redirected binding specificity in their surface proteins are discussed. PMID- 10473427 TI - Enhanced (+)-catechin transglucosylating activity of Streptococcus mutans GS-5 glucosyltransferase-D due to fructose removal. AB - The (+)-catechin transglucosylating activities of several glucosyltransferases (GTFs) from the genus Streptococcus were compared. For this purpose, a mixture of four GTFs from Streptococcus sobrinus SL-1 and recombinant GTF-B and GTF-D from Streptococcus mutans GS-5 expressed in Escherichia coli were studied. It was shown that after removal of alpha-glucosidase activity, GTF-D transglucosylated catechin with the highest efficiency. A maximal yield (expressed as the ratio of moles of glucoside formed to moles of catechin initially added) of 90% was observed with 10 mM catechin and 100 mM sucrose (K(m), 13 mM) in 125 mM potassium phosphate, pH 6.0, at 37 degrees C. (1)H and (13)C nuclear magnetic resonance spectroscopy revealed the structures of two catechin glucosides, (+)-catechin-4' O-alpha-D-glucopyranoside and (+)-catechin-4',7-O-alpha-di-D-glucopyranoside. Fructose accumulation during glucosyl transfer from sucrose to the acceptor competitively inhibited catechin transglucosylation (K(i), 9.3 mM), whereas glucose did not inhibit catechin transglucosylation. The addition of yeasts was studied in order to minimize fructose inhibition by means of fructose removal. For this purpose, the yeasts Pichia pastoris and the mutant Saccharomyces cerevisiae T2-3D were selected because of their inabilities to utilize sucrose. Addition of P. pastoris or S. cerevisiae T2-3D to the standard reaction mixture resulted in a twofold increase in the duration of the maximum GTF-D transglucosylation rate. The addition of the yeasts also stimulated sucrose utilization by GTF-D. PMID- 10473428 TI - Synergistic actions of nisin, sublethal ultrahigh pressure, and reduced temperature on bacteria and yeast. AB - Nisin in combination with ultrahigh-pressure treatment (UHP) showed strong synergistic effects against Lactobacillus plantarum and Escherichia coli at reduced temperatures (<15 degrees C). The strongest inactivation effects were observed when nisin was present during pressure treatment and in the recovery medium. Elimination (>6-log reductions) of L. plantarum was achieved at 10 degrees C with synergistic combinations of 0.5 microg of nisin per ml at 150 MPa and 0.1 microg of nisin per ml at 200 MPa for 10 min. Additive effects of nisin and UHP accounted for only 1.2- and 3.7-log reductions, respectively. Elimination was also achieved for E. coli at 10 degrees C with nisin present at 2 microg/ml, and 10 min of pressure at 200 MPa, whereas the additive effect accounted for only 2.6-log reductions. Slight effects were observed even against the yeast Saccharomyces cerevisiae with nisin present at 5 microg/ml and with 200 MPa of pressure. Combining nisin, UHP, and lowered temperature may allow considerable reduction in time and/or pressure of UHP treatments. Kill can be complete without the frequently encountered survival tails in UHP processing. The slightly enhanced synergistic kill with UHP at reduced temperatures was also observed for other antimicrobials, the synthetic peptides MB21 and histatin 5. The postulated mode of action was that the reduced temperature and the binding of peptides to the membrane increased the efficacy of UHP treatment. The increases in fatty acid saturation or diphosphatidylglycerol content and the lysylphosphatidyl content of the cytoplasm membrane of L. plantarum were correlated with increased susceptibility to UHP and nisin, respectively. PMID- 10473429 TI - Use of molecular and isotopic techniques to monitor the response of autotrophic ammonia-oxidizing populations of the beta subdivision of the class proteobacteria in arable soils to nitrogen fertilizer. AB - This study examined the effects of NH(4)NO(3) fertilizer on the size and activity of nitrifying, autotrophic, ammonia-oxidizing populations of the beta subdivision of the class Proteobacteria in arable soils. Plots under different long-term fertilizer regimes were sampled before and after NH(4)NO(3) additions, and the rates of nitrification were determined by (15)N isotopic pool dilution assays. Ammonia-oxidizing populations in the plots were quantified by competitive PCR assays based on the amoA and ribosomal 16S genes. Prior to fertilizer addition, ammonium concentrations and nitrification rates in the plots were comparatively low; ammonia-oxidizing populations were present at 10(4) to 10(5) gene copies g of soil(-1). Three days after the application of fertilizer, nitrification rates had risen considerably but the size of the ammonia-oxidizing population was unchanged. Six weeks after fertilizer treatment, ammonium concentrations and nitrification rates had fallen while the ammonia-oxidizing populations in plots receiving fertilizer had increased. The rapidity of the rise in nitrification rates observed after 3 days suggests that it results from phenotypic changes in the ammonia-oxidizing bacterial population. Associated increases in population sizes were only observed after 6 weeks and did not correlate directly with nitrifying activity. Phylogenetic analyses of PCR products from one of the plots revealed a population dominated by Nitrosospira-type organisms, similar to those prevalent in other soils. PMID- 10473430 TI - Expression of the isoamylase gene of Flavobacterium odoratum KU in Escherichia coli and identification of essential residues of the enzyme by site-directed mutagenesis. AB - The isoamylase gene from Flavobacterium odoratum KU was cloned into and expressed in Escherichia coli JM109. The promoter of the gene was successful in E. coli, and the enzyme produced was excreted into the culture medium, depending on the amount of the enzyme expressed. The enzyme found in the culture medium showed almost the same M(r), heat-inactivating constant, and N-terminal sequence as those of the enzyme accumulated in the periplasmic space. This result indicated that the enzyme accumulated in an active form at the periplasm was transported out of the cell. The primary sequence of the enzyme, which was deduced from its nucleotide sequence, showed that the mature enzyme consisted of 741 amino acid residues. By changing five possible residues to Ala independently, it was found that Asp-374, Glu-422, and Asp-497 were essential. The sequences around those residues were highly conserved in isoamylases of different origins and the glycogen operon protein X, GlgX. The comparison of the distance between these essential residues with those of various amylases suggested that the bacterial and plant isoamylase but not GlgX had a longer fourth loop than the other amylases. This longer fourth loop had a possible role in accommodating the long branched chains of native glycogens and starches. PMID- 10473431 TI - Development and application of pathovar-specific monoclonal antibodies that recognize the lipopolysaccharide O antigen and the type IV fimbriae of Xanthomonas hyacinthi. AB - The objective of this study was to develop a specific immunological diagnostic assay for yellow disease in hyacinths, using monoclonal antibodies (MAbs). Mice were immunized with a crude cell wall preparation (shear fraction) from Xanthomonas hyacinthi and with purified type IV fimbriae. Hybridomas were screened for a positive reaction with X. hyacinthi cells or fimbriae and for a negative reaction with X. translucens pv. graminis or Erwinia carotovora subsp. carotovora. Nine MAbs recognized fimbrial epitopes, as shown by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy; however, three of these MAbs had weak cross-reactions with two X. translucens pathovars in immunoblotting experiments. Seven MAbs reacted with lipopolysaccharides and yielded a low-mobility ladder pattern on immunoblots. Subsequent analysis of MAb 2E5 showed that it specifically recognized an epitope on the O antigen, which was found to consist of rhamnose and fucose in a 2:1 molar ratio. The cross-reaction of MAb 2E5 with all X. hyacinthi strains tested showed that this O antigen is highly conserved within this species. MAb 1B10 also reacted with lipopolysaccharides. MAbs 2E5 and 1B10 were further tested in ELISA and immunoblotting experiments with cells and extracts from other pathogens. No cross-reaction was found with 27 other Xanthomonas pathovars tested or with 14 other bacterial species from other genera, such as Erwinia and Pseudomonas, indicating the high specificity of these antibodies. MAbs 2E5 and 1B10 were shown to be useful in ELISA for the detection of X. hyacinthi in infected hyacinths. PMID- 10473432 TI - Bacterial growth state distinguished by single-cell protein profiling: does chlorination kill coliforms in municipal effluent? AB - Municipal effluent is the largest reservoir of human enteric bacteria. Its public health significance, however, depends upon the physiological status of the wastewater bacterial community. A novel immunofluorescence assay was developed and used to examine the bacterial growth state during wastewater disinfection. Quantitative levels of three highly conserved cytosolic proteins (DnaK, Dps, and Fis) were determined by using enterobacterium-specific antibody fluorochrome coupled probes. Enterobacterial Fis homologs were abundant in growing cells and nearly undetectable in stationary-phase cells. In contrast, enterobacterial Dps homologs were abundant in stationary-phase cells but virtually undetectable in growing cells. The range of variation in the abundance of both proteins was at least 100-fold as determined by Western blotting and immunofluorescence analysis. Enterobacterial DnaK homologs were nearly invariant with growth state, enabling their use as permeabilization controls. The cellular growth states of individual enterobacteria in wastewater samples were determined by measurement of Fis, Dps, and DnaK abundance (protein profiling). Intermediate levels of Fis and Dps were evident and occurred in response to physiological transitions. The results indicate that chlorination failed to kill coliforms but rather elicited nutrient starvation and a reversible nonculturable state. These studies suggest that the current standard procedures for wastewater analysis which rely on detection of culturable cells likely underestimate fecal coliform content. PMID- 10473433 TI - On the occurrence of anoxic microniches, denitrification, and sulfate reduction in aerated activated sludge. AB - A combination of different methods was applied to investigate the occurrence of anaerobic processes in aerated activated sludge. Microsensor measurements (O(2), NO(2)(-), NO(3)(-), and H(2)S) were performed on single sludge flocs to detect anoxic niches, nitrate reduction, or sulfate reduction on a microscale. Incubations of activated sludge with (15)NO(3)(-) and (35)SO(4)(2-) were used to determine denitrification and sulfate reduction rates on a batch scale. In four of six investigated sludges, no anoxic zones developed during aeration, and consequently denitrification rates were very low. However, in two sludges anoxia in flocs coincided with significant denitrification rates. Sulfate reduction could not be detected in any sludge in either the microsensor or the batch investigation, not even under short-term anoxic conditions. In contrast, the presence of sulfate-reducing bacteria was shown by fluorescence in situ hybridization with 16S rRNA-targeted oligonucleotide probes and by PCR-based detection of genes coding for the dissimilatory sulfite reductase. A possible explanation for the absence of anoxia even in most of the larger flocs might be that oxygen transport is not only diffusional but enhanced by advection, i.e., facilitated by flow through pores and channels. This possibility is suggested by the irregularity of some oxygen profiles and by confocal laser scanning microscopy of the three-dimensional floc structures, which showed that flocs from the two sludges in which anoxic zones were found were apparently denser than flocs from the other sludges. PMID- 10473434 TI - Ti plasmids from Agrobacterium characterize rootstock clones that initiated a spread of crown gall disease in Mediterranean countries. AB - Crown gall caused by Agrobacterium is one of the predominant diseases encountered in rose cultures. However, our current knowledge of the bacterial strains that invade rose plants and the way in which they spread is limited. Here, we describe the integrated physiological and molecular analyses of 30 Agrobacterium isolates obtained from crown gall tumors and of several reference strains. Characterization was based on the determination of the biovar, analysis of 16S ribosomal DNA restriction fragment length polymorphisms by PCR (PCR-RFLP), elucidation of the opine type, and PCR-RFLP analysis of genes involved in virulence and oncogenesis. This study led to the classification of rose isolates into seven groups with common chromosome characteristics and seven groups with common Ti plasmid characteristics. Altogether, the rose isolates formed 14 independent groups, with no specific association of plasmid- and chromosome encoded traits. The predominant Ti plasmid characteristic was that 16 of the isolates induced the production of the uncommon opine succinamopine, while the other 14 were nopaline-producing isolates. With the exception of one, all succinamopine Ti plasmids belonged to the same plasmid group. Conversely, the nopaline Ti plasmids belonged to five groups, one of these containing seven isolates. We showed that outbreaks of disease provoked by the succinamopine producing isolates in different countries and nurseries concurred with a common origin of specific rootstock clones. Similarly, groups of nopaline-producing isolates were associated with particular rootstock clones. These results strongly suggest that the causal agent of crown gall disease in rose plants is transmitted via rootstock material. PMID- 10473435 TI - Detection and identification of amanitins in the wood-rotting fungi Galerina fasciculata and Galerina helvoliceps. AB - More than 600 strains of wood-rotting fungi were screened for the detection of amanitins. Three strains of Galerina fasciculata and 18 strains of Galerina helvoliceps contained amanitins. These strains contained mainly alpha- and beta amanitins in the native fruit bodies, while alpha- and gamma-amanitins were found in liquid-cultured mycelia. Purified amanitins were confirmed by their chromatographic profiles, spectra (UV, Fourier transform infrared, and atmospheric ionization mass), cytotoxicity for mammalian cell lines (3T3 and SiHa), and inhibitory effects on RNA polymerase II. The results revealed that the purified amanitin fractions from these species are identical to authentic amanitins and suggest that these two species must be handled as poisonous mushrooms. PMID- 10473436 TI - Replacement of a metabolic pathway for large-scale production of lactic acid from engineered yeasts. AB - Interest in the production of L-(+)-lactic acid is presently growing in relation to its applications in the synthesis of biodegradable polymer materials. With the aim of obtaining efficient production and high productivity, we introduced the bovine L-lactate dehydrogenase gene (LDH) into a wild-type Kluyveromyces lactis yeast strain. The observed lactic acid production was not satisfactory due to the continued coproduction of ethanol. A further restructuring of the cellular metabolism was obtained by introducing the LDH gene into a K. lactis strain in which the unique pyruvate decarboxylase gene had been deleted. With this modified strain, in which lactic fermentation substituted completely for the pathway leading to the production of ethanol, we obtained concentrations, productivities, and yields of lactic acid as high as 109 g liter(-1), 0.91 g liter(-1) h(-1), and 1.19 mol per mole of glucose consumed, respectively. The organic acid was also produced at pH levels lower than those usual for bacterial processes. PMID- 10473437 TI - Isolation, nucleotide sequence, and physiological relevance of the gene encoding triose phosphate isomerase from Kluyveromyces lactis. AB - Lack of triose phosphate isomerase activity (TIM) is of special interest because this enzyme works at an important branch point of glycolytic flux. In this paper, we report the cloning and sequencing of the Kluyveromyces lactis gene encoding TIM. Unlike Saccharomyces cerevisiae DeltaTPI1 mutants, the K. lactis mutant strain was found to be able to grow on glucose. Preliminary bioconversion experiments indicated that, like the S. cerevisiae TIM-deficient strain, the K. lactis TIM-deficient strain is able to produce glycerol with high yield. PMID- 10473438 TI - Degradation of polycarbonate by a polyester-degrading strain, Amycolatopsis sp. strain HT-6. AB - Amycolatopsis sp. strain HT-6, a poly(tetramethylene succinate) (PTMS)-degrading actinomycete, was observed to degrade poly(tetramethylene carbonate) (PTMC). In a liquid culture with 150 mg of PTMC film, 59% degradation was achieved, but with a low yield of cell growth. On the other hand, PTMS copolymerized with a small amount of PTMC, forming a copolyester carbonate (PEC) that was completely and rapidly degraded with a high yield of cell growth. PMID- 10473439 TI - Isolation of nitrogen-fixing bacteria containing molybdenum-independent nitrogenases from natural environments. AB - Seven diazotrophs that grow well under Mo-deficient, N(2)-fixing conditions were isolated from a variety of environments. These isolates fall in the gamma subdivision of the class Proteobacteria and have genes that encode the Mo nitrogenase (nitrogenase 1) and the V nitrogenase (nitrogenase 2). Four of the isolates also harbor genes that encode the iron-only nitrogenase (nitrogenase 3). PMID- 10473440 TI - Identification of bacterial species associated with the sheep scab mite (Psoroptes ovis) by using amplified genes coding for 16S rRNA. AB - This was the first molecular study of the bacterial flora of the sheep scab mite (Psoroptes ovis). A sequence analysis of genes coding for 16S rRNA revealed that Serratia marcescens and bacteria closely related to Staphylococcus intermedius or Staphylococcus chromogens and Alloiococcus otitidis were present. These bacteria were associated with skin lesions, dermatitis, and otitis media caused by P. ovis. PMID- 10473441 TI - Community size and metabolic rates of psychrophilic sulfate-reducing bacteria in Arctic marine sediments. AB - The numbers of sulfate reducers in two Arctic sediments with in situ temperatures of 2.6 and -1.7 degrees C were determined. Most-probable-number counts were higher at 10 degrees C than at 20 degrees C, indicating the predominance of a psychrophilic community. Mean specific sulfate reduction rates of 19 isolated psychrophiles were compared to corresponding rates of 9 marine, mesophilic sulfate-reducing bacteria. The results indicate that, as a physiological adaptation to the permanently cold Arctic environment, psychrophilic sulfate reducers have considerably higher specific metabolic rates than their mesophilic counterparts at similarly low temperatures. PMID- 10473442 TI - Clostridium difficile cell attachment is modified by environmental factors. AB - Adherence of Clostridium difficile to Vero cells under anaerobic conditions was increased by a high sodium concentration, calcium-rich medium, an acidic pH, and iron starvation. The level of adhesion of nontoxigenic strains was comparable to that of toxigenic strains. Depending on the bacterial culture conditions, Vero cells could bind to one, two, or three bacterial surface proteins with molecular masses of 70, 50, and 40 kDa. PMID- 10473443 TI - Effect of methodology on the tuberculocidal activity of a glutaraldehyde-based disinfectant. AB - Although official guidelines recommend a plate counting method for testing the susceptibility of mycobacteria to disinfectants, manufacturers usually prefer to employ the BACTEC procedure. Data showing that the BACTEC method overestimates the activity of a glutaraldehyde-based disinfectant against Mycobacterium tuberculosis in comparison with a conventional plate counting procedure are presented. PMID- 10473444 TI - Genus- and species-specific PCR-based detection of dairy propionibacteria in environmental samples by using primers targeted to the genes encoding 16S rRNA. AB - PCR assays with primers targeted to the genes encoding 16S rRNA were developed for detection of dairy propionibacteria. Propionibacterium thoenii specific oligonucleotide PT3 was selected after partial resequencing. Tests allowed the detection of less than 10 cells per reaction from milk and cheese and 10(2) cells per reaction from forage and soil. PMID- 10473445 TI - Temperature-dependent fermentation of D-sorbitol in Escherichia coli O157:H7. AB - The influence of growth temperature on the ability to ferment D-sorbitol was investigated in Escherichia coli O157:H7. It was found that O157:H7 strains have a temperature-sensitive sorbitol phenotype. D-Sorbitol transport and sorbitol-6 phosphate dehydrogenase activities were expressed in sorbitol-fermenting cells grown at 30 degrees C but only at a low level at 40 degrees C. Sorbitol-positive variants able to transport D-sorbitol were easily selected at 30 degrees C from culture of Sor(-) E. coli O157:H7 strains. PMID- 10473446 TI - Variation in resistance to hydrostatic pressure among strains of food-borne pathogens. AB - Among food-borne pathogens, some strains could be resistant to hydrostatic pressure treatment. This information is necessary to establish processing parameters to ensure safety of pressure-pasteurized foods (N. Kalchayanand, A. Sikes, C. P. Dunne, and B. Ray, J. Food Prot. 61:425-431, 1998). We studied variation in pressure resistance among strains of Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella species at two temperatures of pressurization. Early-stationary-phase cells in 1% peptone solution were pressurized at 345 MPa either for 5 min at 25 degrees C or for 5, 10, or 15 min at 50 degrees C. The viability loss (in log cycles) following pressurization at 25 degrees C ranged from 0.9 to 3.5 among nine L. monocytogenes strains, 0.7 to 7.8 among seven S. aureus strains, 2.8 to 5.6 among six E. coli O157:H7 strains, and 5.5 to 8.3 among six Salmonella strains. The results show that at 25 degrees C some strains of each species are more resistant to pressure than the others. However, when one resistant and one sensitive strain from each species were pressurized at 345 MPa and 50 degrees C, the population of all except the resistant S. aureus strain was reduced by more than 8 log cycles within 5 min. Viability loss of the resistant S. aureus strain was 6.3 log cycles even after 15 min of pressurization. This shows that strains of food-borne pathogens differ in resistance to hydrostatic pressure (345 MPa) at 25 degrees C, but this difference is greatly reduced at 50 degrees C. Pressurization at 50 degrees C, in place of 25 degrees C, will ensure greater safety of foods. PMID- 10473447 TI - Role of humic-bound iron as an electron transfer agent in dissimilatory Fe(III) reduction. AB - The dissimilatory Fe(III) reducer Geobacter metallireducens reduced Fe(III) bound in humic substances, but the concentrations of Fe(III) in a wide range of highly purified humic substances were too low to account for a significant portion of the electron-accepting capacities of the humic substances. Furthermore, once reduced, the iron in humic substances could not transfer electrons to Fe(III) oxide. These results suggest that other electron-accepting moieties in humic substances, such as quinones, are the important electron-accepting and shuttling agents under Fe(III)-reducing conditions. PMID- 10473448 TI - Bacterial spores survive treatment with commercial sterilants and disinfectants. AB - This study compared the activity of commercial liquid sterilants and disinfectants on Bacillus subtilis spores deposited on three types of devices made of noncorrodible, corrodible, or polymeric material. Products like Renalin, Exspor, Wavicide-01, Cidexplus, and cupric ascorbate were tested under conditions specified for liquid sterilization. These products, at the shorter times indicated for disinfection, and popular disinfectants, like Clorox, Cavicide, and Lysol were also studied. Data obtained with a sensitive and quantitative test suggest that commercial liquid sterilants and disinfectants are less effective on contaminated surfaces than generally acknowledged. PMID- 10473449 TI - Perkinsus marinus extracellular protease modulates survival of Vibrio vulnificus in Eastern oyster (Crassostrea virginica) hemocytes. AB - The in vitro effects of the Perkinsus marinus serine protease on the intracellular survival of Vibrio vulnificus in oyster hemocytes were examined by using a time-course gentamicin internalization assay. Results showed that protease-treated hemocytes were initially slower to internalize V. vulnificus than untreated hemocytes. After 1 h, the elimination of V. vulnificus by treated hemocytes was significantly suppressed compared with hemocytes infected with invasive and noninvasive controls. Our data suggest that the serine protease produced by P. marinus suppresses the vibriocidal activity of oyster hemocytes to effectively eliminate V. vulnificus, potentially leading to conditions favoring higher numbers of vibrios in oyster tissues. PMID- 10473450 TI - Identification of Lactobacillus isolates from the gastrointestinal tract, silage, and yoghurt by 16S-23S rRNA gene intergenic spacer region sequence comparisons. AB - Lactobacillus isolates were identified by PCR amplification and sequencing of the region between the 16S and 23S rRNA genes (spacer region). The sequences obtained from the isolates were compared to those of reference strains held in GenBank. A similarity of 97.5% or greater was considered to provide identification. To check the reliability of the method, the V2-V3 region of the 16S rRNA gene was amplified and sequenced in the case of isolates whose spacer region sequences were less than 99% similar to that of a reference strain. Confirmation of identity was obtained in all instances. Spacer region sequencing provided rapid and accurate identification of Lactobacillus isolates obtained from gastrointestinal, yoghurt, and silage samples. It had an advantage over 16S V2-V3 sequence comparisons because it distinguished between isolates of Lactobacillus casei and Lactobacillus rhamnosus. PMID- 10473451 TI - Poly(aspartic acid) degradation by a Sphingomonas sp. isolated from freshwater. AB - A poly(aspartic acid) degrading bacterium (strain KT-1 [JCM10459]) was isolated from river water and identified as a member of the genus Sphingomonas. The isolate degraded only poly(aspartic acid)s of low molecular masses (<5 kDa), while the cell extract hydrolyzed high-molecular-mass poly(aspartic acid)s of 5 to 150 kDa to yield aspartic acid monomer. PMID- 10473452 TI - Diversity and heterogeneity of epibiotic bacterial communities on the marine nematode Eubostrichus dianae. AB - The diversity of a microbial community covering the surface of a marine nematode was analyzed by performing a 16S ribosomal DNA (rDNA) restriction cutting and sequencing analysis. In two clone libraries constructed by using individual nematodes, 54 and 85 restriction patterns were identified, and only 13 of these patterns were common to both libraries. Sequence analysis indicated that the common patterns belonged to four groups related to sequences of cytophagas, sulfate-reducing bacteria, members of the gamma subclass of the class Proteobacteria, and caulobacters. At least two groups appeared to be permanent members of the community as they were also detected in a 16S rDNA library constructed 3 years previously by using 100 pooled nematode specimens. A surprising outcome was that very dominant filamentous bacteria were apparently not represented in the clone libraries, as quantitative probing showed that none of the common operational taxonomic unit groups displayed the expected overwhelming dominance. Nevertheless, our analysis revealed both an unexpectedly high level of bacterial diversity and heterogeneity in samples representing presumably very similar microenvironments. PMID- 10473453 TI - Efficacy of electrolyzed oxidizing water for inactivating Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes. AB - The efficacy of electrolyzed oxidizing water for inactivating Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes was evaluated. A five strain mixture of E. coli O157:H7, S. enteritidis, or L. monocytogenes of approximately 10(8) CFU/ml was inoculated in 9 ml of electrolyzed oxidizing water (treatment) or 9 ml of sterile, deionized water (control) and incubated at 4 or 23 degrees C for 0, 5, 10, and 15 min; at 35 degrees C for 0, 2, 4, and 6 min; or at 45 degrees C for 0, 1, 3, and 5 min. The surviving population of each pathogen at each sampling time was determined on tryptic soy agar. At 4 or 23 degrees C, an exposure time of 5 min reduced the populations of all three pathogens in the treatment samples by approximately 7 log CFU/ml, with complete inactivation by 10 min of exposure. A reduction of >/=7 log CFU/ml in the levels of the three pathogens occurred in the treatment samples incubated for 1 min at 45 degrees C or for 2 min at 35 degrees C. The bacterial counts of all three pathogens in control samples remained the same throughout the incubation at all four temperatures. Results indicate that electrolyzed oxidizing water may be a useful disinfectant, but appropriate applications need to be validated. PMID- 10473454 TI - Detection of Verrucomicrobia in a pasture soil by PCR-mediated amplification of 16S rRNA genes. AB - Oligonucleotide primers were designed and used to amplify, by PCR, partial 16S rRNA genes of members of the bacterial division Verrucomicrobia in DNA extracted from a pasture soil. By applying most-probable-number theory to the assay, verrucomicrobia appeared to contribute some 0.2% of the soil DNA. Amplified ribosomal DNA restriction analysis of 53 cloned PCR-amplified partial 16S rRNA gene fragments and comparative sequence analysis of 21 nonchimeric partial 16S rRNA genes showed that these primers amplified only 16S rRNA genes of members of the Verrucomicrobia in DNA extracted from the soil. PMID- 10473455 TI - Impact of rpoS deletion on Escherichia coli biofilms. AB - Slow growth has been hypothesized to be an essential aspect of bacterial physiology within biofilms. In order to test this hypothesis, we employed two strains of Escherichia coli, ZK126 (DeltalacZ rpoS(+)) and its isogenic DeltarpoS derivative, ZK1000. These strains were grown at two rates (0.033 and 0.0083 h( 1)) in a glucose-limited chemostat which was coupled either to a modified Robbins device containing plugs of silicone rubber urinary catheter material or to a glass flow cell. The presence or absence of rpoS did not significantly affect planktonic growth of E. coli. In contrast, biofilm cell density in the rpoS mutant strain (ZK1000), as measured by determining the number of CFU per square centimeter, was reduced by 50% (P < 0.05). Deletion of rpoS caused differences in biofilm cell arrangement, as seen by scanning confocal laser microscopy. In reporter gene experiments, similar levels of rpoS expression were seen in chemostat-grown planktonic and biofilm populations at a growth rate of 0.033 h( 1). Overall, these studies suggest that rpoS is important for biofilm physiology. PMID- 10473456 TI - Characterization of two Bacillus probiotics. AB - Bacillus subtilis is currently used as an oral probiotic. We examined two commercial B. subtilis probiotic preparations, Enterogermina and Biosubtyl. Surprisingly, physiological and genetic characterization of the bacteria contained in each of these preparations has shown that neither contains B. subtilis. PMID- 10473457 TI - Association of bacteria and yeasts in hot springs. AB - The thermophilic bacterium Bacillus sp. strain TB-1 was isolated in association with the yeast Debaryomyces vanriji from hot springs at 46 degrees C. It was shown that TB-1 excreted thiamine into the culture broth, which not only promoted D. vanriji growth in mixed culture but also increased the maximal temperature for yeast growth. PMID- 10473458 TI - Phantoms in the brain. Question the assumption that the adult brain is "hard wired". PMID- 10473459 TI - Pituitary surgery for acromegaly. Should be done by specialists. PMID- 10473461 TI - Incontinence campaign tests limits of advertising rules. PMID- 10473460 TI - British guidelines on managing hypertension. Provide evidence, progress, and an occasional missed opportunity. PMID- 10473462 TI - Public awareness of hypertension declines in US PMID- 10473463 TI - Israel introduces generic drugs scheme. PMID- 10473464 TI - Turkey surprised by extent of aid. PMID- 10473465 TI - In brief PMID- 10473466 TI - Record numbers of Irish women visit Britain for abortion. PMID- 10473467 TI - Dolly's other DNA came from donor egg PMID- 10473468 TI - South African doctors demand action on "unethical" colleagues. PMID- 10473469 TI - Canadian provinces want to cut tests to meet doctor shortage. PMID- 10473470 TI - Romanian GPs to take control of their practices PMID- 10473471 TI - The effectiveness of glucocorticoids in treating croup: meta-analysis. AB - OBJECTIVE: To determine the effectiveness of glucocorticoid treatment in children with croup. DESIGN: Meta-analysis of randomised controlled trials that examine the effectiveness of glucocorticoid treatment in children with croup. MAIN OUTCOME MEASURES: Score on scale measuring severity of croup, use of cointerventions (adrenaline (epinephrine), antibiotics, or supplemental glucocorticoids), length of stay in accident and emergency or in hospital, and rate of hospitalisation. RESULTS: Twenty four studies met the inclusion criteria. Glucocorticoid treatment was associated with an improvement in the croup severity score at 6 hours with an effect size of -1.0 (95% confidence interval -1.5 to 0.6) and at 12 hours -1.0 (-1.6 to -0.4); at 24 hours this improvement was no longer significant (-1.0, -2.0 to 0.1). There was a decrease in the number of adrenaline treatments needed in children treated with glucocorticoids: a decrease of 9% (95% confidence interval 2% to 16%) among those treated with budesonide and of 12% (4% to 20%) among those treated with dexamethasone. There was also a decrease in the length of time spent in accident and emergency (-11 hours, 95% confidence interval -18 to 4 hours), and for inpatients hospital stay was reduced by 16 hours (-31 to 1 hour). Publication bias seems to play a part in these results. CONCLUSIONS: Dexamethasone and budesonide are effective in relieving the symptoms of croup as early as 6 hours after treatment. Fewer cointerventions are used and the length of time spent in hospital is decreased in patients treated with glucocorticoids. PMID- 10473472 TI - Predictors and consequences of unemployment among construction workers: prospective cohort study. AB - OBJECTIVE: To study predictors and consequences of unemployment. DESIGN: Prospective cohort study. SETTING: 11 construction companies in southern Finland. PARTICIPANTS: 586 male employees, aged 40-59 years at baseline in 1991 and not retired during a 4 year follow up. MAIN OUTCOME MEASURES: Long term unemployment, stress symptoms, disease, alcohol consumption, exercise activity, and body mass index. RESULTS: In a multiple logistic regression model, long term unemployment (>24 months v /=75 mg. Further research is required to evaluate the effect of this approach in combination with appropriate targeting of treatment and advice about dosage. PMID- 10473481 TI - Claude bernard on the action of curare PMID- 10473480 TI - Identifying problems with data collection at a local level: survey of NHS maternity units in England. AB - OBJECTIVES: To document the extent to which maternity data are collected and how they are recorded, and to identify problems that may affect their availability throughout the NHS. METHODS: Postal survey in September 1997 with structured questionnaires. SETTING: 207 NHS trusts with maternity units in England. PARTICIPANTS: Heads of midwifery in maternity units. MAIN OUTCOME MEASURES: Extent to which maternity data were routinely recorded, how they were recorded and evaluated, and to whom they were made available. RESULTS: 167 (81%) of questionnaires were returned, representing 166 trusts. Of these trusts, 165 collected >/=17 of the 19 data items in HES maternity tail, and 158 collected >/=40 of the 45 items selected from Korner dataset. Only 18 collected all five items selected from the "indicators of success," and 17 did not collect any. In 58 of trusts data were primarily recorded on paper. A computerised maternity information system was used by 106 (63%) of trusts, but many recorded data on paper first. Thirty four did not audit data for accuracy. Most trusts analysed data not routinely collected at national level, but 18 did not analyse HES maternity tail and 17 did not analyse Korner data. CONCLUSIONS: Improvement is needed in quality, completeness, and availability of maternity data at a national level, particularly if the NHS information strategy is to be successfully implemented. Although most of the data items in national datasets are recorded locally, variations in the way data are defined, recorded, and analysed and lack of linkage between computer systems restrict their access, availability, and use at local, district, and national levels. PMID- 10473483 TI - Morphine induced allodynia in a child with brain tumour. PMID- 10473482 TI - Acute dystonia induced by drug treatment. PMID- 10473484 TI - Diversity to divinity PMID- 10473485 TI - British Hypertension Society guidelines for hypertension management 1999: summary. PMID- 10473486 TI - Handling uncertainty in economic evaluations of healthcare interventions. PMID- 10473487 TI - How the defibrillator saved a patient's life PMID- 10473488 TI - When doctors might kill their patients. Palliative care physicians always have their patients' best interests in mind. PMID- 10473489 TI - Increased mortality from liver cancer in England and Wales is not related to hepatitis C. PMID- 10473490 TI - Effectiveness of rivastigmine in Alzheimer's disease. Improvements in functional ability remain unestablished. PMID- 10473491 TI - Screening and mortality from cervical cancer. Does screening really reduce mortality? PMID- 10473492 TI - Use of guidelines should be evaluated in randomised controlled trials. PMID- 10473493 TI - Cognitive therapy is no better than supportive counselling in schizophrenia. PMID- 10473494 TI - Door to needle times of 12 minutes are possible in one emergency department. PMID- 10473495 TI - Early trials of angiogenic factors have not targeted patients most at risk of ocular disease. PMID- 10473497 TI - Peter thomas fallon PMID- 10473496 TI - Smoking by parents of asthmatic children. Sensitive counselling may still be worth while. PMID- 10473498 TI - Menopause and hormone replacement therapy: effective patient care PMID- 10473500 TI - Doctors from hell PMID- 10473499 TI - Terminal consent PMID- 10473501 TI - Earthquake disaster in turkey PMID- 10473502 TI - My years with lyme disease PMID- 10473503 TI - Assets for the meritocracy PMID- 10473505 TI - Long term unemployed workers improve their lifestyle PMID- 10473504 TI - Glucocorticoids are effective in treating croup PMID- 10473506 TI - Senior doctors do not use their GP PMID- 10473507 TI - Critically ill children show same immunoparalysis as adults PMID- 10473508 TI - Women in ethnic minorities are less likely to take HRT PMID- 10473509 TI - Drug counselling improves adherence to antidepressants PMID- 10473510 TI - Clinical utility of testing for antineutrophil cytoplasmic antibodies. PMID- 10473511 TI - Proposed mechanisms of transfusion-induced immunomodulation. PMID- 10473512 TI - Interleukin 4 in systemic sclerosis: not just an increase. PMID- 10473513 TI - Increased levels of alternatively spliced interleukin 4 (IL-4delta2) transcripts in peripheral blood mononuclear cells from patients with systemic sclerosis. AB - Recent in vitro studies have shown that interleukin 4 (IL-4) induces and gamma interferon (IFN-gamma) inhibits collagen production. To define the TH1(IFN-gamma) and TH2(IL-4) cytokine profiles in systemic sclerosis (Sscl), a disease characterized by widespread fibrosis, we investigated IL-4 and IFN-gamma transcripts in peripheral blood mononuclear cells and plasma protein levels in 13 patients with Sscl. Two previously identified IL-4 transcripts, a full-length transcript and an alternatively spliced (truncated) transcript (designated IL 4delta2), were identified in patients and normal controls. Significantly increased levels of total IL-4 transcripts (full-length plus IL-4delta2 transcripts) were found in patients with Sscl in comparison to those found in healthy controls (P = 0.003), and this increase was primarily due to an increase in the level of the alternatively spliced IL-4delta2 form. The IL-4delta2/full length-IL-4 transcript ratio was significantly increased in Sscl patients (P < 0.0001, versus healthy controls). Sequencing analysis revealed that the frequency of IL-4 clones carrying the IL-4delta2 transcript was also substantially increased in patients with Sscl. Plasma IL-4 protein levels were increased in Sscl patients compared to those in healthy controls (P = 0.001) and correlated with total IL-4 transcript levels. The up-regulation of the fibrogenic IL-4 (a TH2 cytokine) in Sscl suggests a pathogenic role for IL-4 in this disease. PMID- 10473514 TI - Low levels of antigenic variability in fluconazole-susceptible and -resistant Candida albicans isolates from human immunodeficiency virus-infected patients with oropharyngeal candidiasis. AB - Three serial isolates of Candida albicans were obtained by direct swab or by oral saline rinses from each of five human immunodeficiency virus-infected patients with recurrent oropharyngeal candidiasis. Genotyping techniques confirmed the presence of a persistent strain in multiple episodes from the same patient, which was different from the strains isolated from other patients. Fluconazole susceptibility was determined by both an agar dilution method and the National Committee for Clinical Laboratory Standards macrobroth procedure. In four of these patients the strains developed fluconazole resistance, and in one patient the strain remained susceptible. The different isolates were propagated as yeast cells on a synthetic medium, and their cell wall proteinaceous components were extracted by treatment with beta-mercaptoethanol. Protein and mannoprotein components present in the extracts were analyzed by electrophoresis, immunoblotting, and lectin-blotting techniques. The analysis showed a similar composition, with only minor qualitative and quantitative differences in the polypeptidic and antigenic patterns associated with the cell wall extracts from serial isolates from the same patient, as well as those from different strains isolated from different patients. Use of monospecific antibodies generated against two immunodominant antigens during candidiasis (enolase and the 58-kDa fibrinogen-binding mannoprotein) demonstrated their expression in all isolates tested. Overall, the antigenic makeup of C. albicans strains remained constant during the course of infection and was not affected by development of fluconazole resistance. In contrast to previous reports, the low degree of antigenic variability observed in this study may be due to the fact that the isolates were obtained from a highly homogeneous population of patients and to the uniformity in techniques used for the isolation, storage, and culture of the different strains, as well as extraction methodologies. PMID- 10473515 TI - Mastitis and immunological factors in breast milk of lactating women in Malawi. AB - Although an elevated sodium concentration in human milk is suggested to be an indicator of mastitis, it is unclear whether elevated sodium concentrations are associated with immunological and inflammatory mediators in human milk. We conducted a cross-sectional study to evaluate the relationships between elevated breast milk sodium concentrations and levels of lactoferrin, lysozyme, secretory leukocyte protease inhibitor (SLPI), interleukin-8 (IL-8), and RANTES (regulated on activation normal T cell expressed and secreted) in human milk at 6 weeks postpartum in 96 lactating women in Blantyre, Malawi. Mastitis, as indicated by an elevated breast milk sodium concentration, was present in 15.6% of the women. Women with and without mastitis had respective median levels of other factors as follows: lactoferrin, 1,230 versus 565 mg/liter (P < 0. 0007); lysozyme, 266 versus 274 mg/liter (P = 0.55); SLPI, 76 versus 15 microg/liter, (P < 0.0002); IL 8, 339 versus 25 ng/liter (P < 0. 0001); and RANTES, 82 versus 3 ng/liter (P < 0.0001). Elevated sodium concentrations in breast milk are associated with an increase in levels of some immunological and inflammatory factors in breast milk. PMID- 10473516 TI - Development of diagnostic reagents to differentiate between Mycobacterium bovis BCG vaccination and M. bovis infection in cattle. AB - In Great Britain a recent independent scientific review for the government has concluded that the development of a cattle vaccine against Mycobacterium bovis holds the best long-term prospect for tuberculosis control in British herds. A sine qua non for vaccination is the development of a complementary diagnostic test to differentiate between vaccinated animals and those infected with M. bovis so that test-and-slaughter-based control strategies can continue alongside vaccination. In order to assess the feasibility of developing a differential diagnostic test for a live vaccine, we chose M. bovis BCG Pasteur as a model system. Recombinant forms of antigens which are expressed in M. bovis but not, or only at low levels, in BCG Pasteur (ESAT-6, MPB64, MPB70, and MPB83) were produced. These reagents were tested either alone or in combination by using peripheral blood mononuclear cells from M. bovis-infected, BCG-vaccinated, and Mycobacterium avium-sensitized calves. All four antigens induced in vitro proliferation and gamma interferon responses only in M. bovis-infected animals. A cocktail composed of ESAT-6, MPB64, and MPB83 identified infected animals but not those vaccinated with BCG. In addition, promiscuous T-cell epitopes of ESAT-6, MPB64, and MPB83 were formulated into a peptide cocktail. In T-cell assays with this peptide cocktail, infected animals were identified with frequencies similar to those obtained in assays with the protein cocktail, while BCG-vaccinated or M. avium-sensitized animals did not respond. In summary, our results suggest that peptide and protein cocktails can be designed to discriminate between M. bovis infection and BCG vaccination. PMID- 10473517 TI - Urinary excretion of thiol compounds in patients with rheumatoid arthritis. AB - The objective of the present study was to assess the excretion of urinary thiol compounds in patients with active and inactive rheumatoid arthritis (RA). Urinary thiol compounds were measured by the method of Kokonov (M. T. Kokonov, Lab. Delo 5:273-276, 1965) in 51 outpatients with active and inactive RA. Those with active disease had significantly higher levels of urinary thioamine excretion. PMID- 10473518 TI - Dot blot assay for detection of antidiacyltrehalose antibodies in tuberculous patients. AB - A simple dot blot test with diacyltrehalose (DAT) as the antigen was developed to detect anti-DAT antibodies in tuberculous patients. To enhance antigen-antibody reaction detection, rabbit serum raised against human immunoglobulins was used prior to incubation with a protein A-colloidal gold complex. With the dot blot system, it was possible to obtain a sensitivity similar to that of enzyme-linked immunosorbent assay (ELISA) and a specificity of 97.14%, versus a specificity of 94.29% by the ELISA. We conclude that this simple and fast assay could be used in places where ELISA equipment is not easy available and that it might also be applicable with other Mycobacterium tuberculosis immunogenic antigens. PMID- 10473520 TI - Detection of borna disease virus-reactive antibodies from patients with psychiatric disorders and from horses by electrochemiluminescence immunoassay. AB - The prevalence of Borna disease virus (BDV)-specific antibodies among patients with psychiatric disorders and healthy individuals has varied in several reports using several different serological assay methods. A reliable and specific method for anti-BDV antibodies needs to be developed to clarify the pathological significance of BDV infections in humans. We developed a new electrochemiluminescence immunoassay (ECLIA) for the antibody to BDV that uses two recombinant proteins of BDV, p40 and p24 (full length). Using this ECLIA, we examined 3,476 serum samples from humans with various diseases and 917 sera from blood donors in Japan for the presence of anti-BDV antibodies. By ECLIA, 26 (3.08%) of 845 schizophrenia patients and 9 (3.59%) of 251 patients with mood disorders were seropositive for BDV. Among 323 patients with other psychiatric diseases, 114 with neurological diseases, 75 with chronic fatigue syndrome, 85 human immunodeficiency virus-infected patients, 50 with autoimmune diseases including rheumatoid arthritis and systemic lupus erythematosis and 17 with leprosy, there was no positive case except one case each with alcohol addiction, AIDS, and dementia. Although 19 (1.36%) of 1,393 patients with various ocular diseases, 10 (1.09%) of 917 blood donors, and 3 (4.55%) of 66 multitransfused patients were seropositive for BDV-specific antigen, high levels of seroprevalence in schizophrenia patients and young patients (16 to 59 years old) with mood disorders were statistically significant. The immunoreactivity of seropositive sera could be verified for specificity by blocking with soluble p40 and/or p24 recombinant protein. Anti-p24 antibody was more frequent than p40 antibody in most cases, and in some psychotic patients antibody profiles showed only p40 antibody. Although serum positive for both p40 and p24 antibodies was not found in this study, the p40 ECLIA count in schizophrenia patients was higher than that of blood donors. Furthermore, we examined 90 sera from Japanese feral horses. Antibody profiles of control human samples are similar to that of naturally BDV-infected feral horses. We concluded that BDV infection was associated in some way with psychiatric disorders. PMID- 10473519 TI - Immune response of children who develop persistent diarrhea following rotavirus infection. AB - A prospective study was conducted with Bangladeshi children with rotavirus (RV) diarrhea to assess whether nutritional and clinical parameters, RV serotypes, levels of interleukin-10 (IL-10), tumor necrosis factor alpha (TNF-alpha), and gamma interferon (IFN-gamma), and RV-specific antibody titers in plasma and stool were associated with the development of persistent diarrhea. Children with watery diarrhea for 6 to 8 days, selected from the Dhaka Hospital of the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B), were enrolled in the study and monitored until diarrhea improved. Children were classified as having acute diarrhea (AD) if diarrhea resolved within 14 days of onset and as having persistent diarrhea (PD) if diarrhea persisted for more than 14 days after onset. Uninfected, control children (n = 13) from the Nutrition Follow-Up Unit of ICDDR,B were also enrolled. Of the 149 children with diarrhea enrolled, 29 had diarrhea with RV alone, of which 19 had AD and 10 developed PD. Samples of stool and blood were collected from all children on enrollment. Stool samples were collected again from children when they developed PD. Of the 10 children who had an initial RV infection and then developed PD, only one had persistent RV infection. Plasma levels of IL-10 and TNF-alpha were higher in children with diarrhea compared to uninfected controls but were similar in children with AD and PD. Plasma IFN-gamma levels were higher in children who developed PD than in those with AD (P = 0.008) or uninfected controls (P = 0.001). In stools, the levels of TNF-alpha, the only cytokine detected, were similar in the three groups of children. RV-specific immunoglobulin G (IgG) titers in plasma were higher in uninfected children than in those with AD (P < 0.001) or PD (P = 0.024) but titers were similar in children with AD and PD. RV-specific IgA titers in plasma and stool were similar in the three groups of children. From all observed parameters, only elevated plasma IFN-gamma levels were associated with subsequent development of PD. However, a larger sample size is necessary to substantiate this observation. PMID- 10473521 TI - A reverse-sandwich enzyme-linked immunosorbent assay for verocytotoxin 1 and 2 antibodies in human and bovine sera. AB - A reverse-sandwich enzyme-linked immunosorbent assay (ELISA), in which an antibody is sandwiched by antigens, was established for the titration of antibodies to verocytotoxins (VT) in human and animal sera. This assay has two advantages over a conventional indirect ELISA: (i) higher specificity and sensitivity and (ii) the ability to comparably titrate antibodies from different species. The VT1 (Shiga-like toxin 1) antibody-positive rates were 5% in 202 normal adult humans and 99% in 93 normal cattle at a dairy farm. This ELISA is most suitable for seroepidemiologic studies of infections with VT-producing Escherichia coli in humans and various animal species. PMID- 10473522 TI - Comparison of PanBio dengue duo enzyme-linked immunosorbent assay (ELISA) and MRL dengue fever virus immunoglobulin M capture ELISA for diagnosis of dengue virus infections in Southeast Asia. AB - The performances of the MRL dengue fever virus immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) and the PanBio Dengue Duo IgM capture and IgG capture ELISA were compared. Eighty sera from patients with dengue virus infections, 24 sera from patients with Japanese encephalitis (JE), and 78 sera from patients with nonflavivirus infections, such as malaria, typhoid, leptospirosis, and scrub typhus, were used. The MRL test showed superior sensitivity for dengue virus infections (94 versus 89%), while the PanBio test showed superior specificity for JE (79 versus 25%) and other infections (100 versus 91%). The PanBio ELISA showed better overall performance, as assessed by the sum of sensitivity and specificity (F value). When dengue virus and nonflavivirus infections were compared, F values of 189 and 185 were obtained for the PanBio and MRL tests, respectively, while when dengue virus infections and JE were compared, F values of 168 and 119 were obtained. The results obtained with individual sera in the PanBio and MRL IgM ELISAs showed good correlation, but this analysis revealed that the cutoff value of the MRL test was set well below that of the PanBio test. Comparing the sensitivity and specificity of the tests at different cutoff values (receiver-operator analysis) revealed that the MRL and PanBio IgM ELISAs performed similarly in distinguishing dengue virus from nonflavivirus infections, although the PanBio IgM ELISA showed significantly better distinction between dengue virus infections and JE. The implications of these findings for the laboratory diagnosis of dengue are discussed. PMID- 10473523 TI - Modulation of T-cell responses to a recall antigen in human T-cell leukemia virus type 1-infected individuals. AB - To determine the mechanism of the purified protein derivative (PPD)-specific hyporesponsiveness in Mycobacterium bovis BCG-vaccinated human T-cell leukemia virus type 1 (HTLV-1)-infected individuals, we examined cytokine production in response to PPD in the following four groups of individuals: (i) HTLV-negative, PPD nonresponders (n = 11; NN); (ii) HTLV-negative, PPD responders (n = 18; NP); (iii) HTLV-positive, PPD nonresponders (n = 15; PN); and (iv) HTLV-positive, PPD responders (n = 15; PP). In vitro stimulation with PPD resulted in both proliferative responses and gamma interferon (IFN-gamma) production in NP and PP (P < 0.02), with minimal proliferation and IFN-gamma production in the NN and PN groups. Further, PPD-specific interleukin 10 (IL-10) production was significantly reduced in the PN group (P < 0.01), while the other groups had comparable levels. Cytokine reconstitution experiments demonstrated that while addition of recombinant IL-12 (rIL-12) plus anti-IL-4 restored PPD-specific responses in the NN group, it had no effect in the PN group. However, addition of rIL-12 resulted in the increased production of IFN-gamma in both nonresponder groups (NN and PN), suggesting that the lack of IFN-gamma production was not responsible for the PPD anergy. We conclude that PPD-specific anergy in HTLV-1-infected individuals appears to be due in part to their inability to respond to rIL-12. PMID- 10473525 TI - A new method with general diagnostic utility for the calculation of immunoglobulin G avidity. AB - The reference method for immunoglobulin G (IgG) avidity determination includes reagent-consuming serum titration. Aiming at better IgG avidity diagnostics, we applied a logistic model for the reproduction of antibody titration curves. This method was tested with well-characterized serum panels for cytomegalovirus, Epstein-Barr virus, rubella virus, parvovirus B19, and Toxoplasma gondii. This approach for IgG avidity calculation is generally applicable and attains the diagnostic performance of the reference method while being less laborious and twice as cost-effective. PMID- 10473524 TI - Deficient antibody-dependent cellular cytotoxicity against human immunodeficiency virus (HIV)-expressing target cells in perinatal HIV infection. AB - Peripheral blood mononuclear cells (PBMC) of human immunodeficiency virus (HIV) infected children, age-matched HIV-seronegative controls, and HIV-infected asymptomatic and symptomatic adults were compared for their ability to mediate antibody-dependent cellular cytotoxicity (ADCC) and natural killer (NK) cell mediated cytotoxicity against target cells expressing HIV or herpes simplex virus (HSV) antigens. Target cells consisted of CD4 lymphocytes purified from PBMC of HIV-seronegative adults and incubated with the IIIB strain of HIV, HUT78 cells chronically infected with IIIB, and HSV-infected human fibroblasts. PBMC of asymptomatic HIV-infected adults were generally able to lyse CD4 cells expressing HIV antigens. Direct correlation was found between the magnitude of lysis and absolute CD4 cell counts in these individuals. In contrast to these results, PBMC from HIV-infected children were generally unable to lyse IIIB-expressing CD4 cells, regardless of the children's clinical status, age, or absolute CD4 cell counts. Cells from HIV-seronegative adults and children did not directly lyse these target cells either but, in contrast to cells of HIV-seropositive children, were able to mediate cell lysis when serum from an HIV-seropositive adult was added. However, effector cells from these HIV-infected children were able to mediate both ADCC against HSV-infected fibroblasts and NK cell-mediated cytotoxicity against IIIB-infected HUT78 cells. Reduced ability of PBMC from vertically HIV-infected children to mediate ADCC against HIV antigen-expressing CD4 cells may contribute to rapid progression to AIDS. PMID- 10473526 TI - Cell culture of sporadic hepatitis E virus in China. AB - The isolation and identification of the 87A strain of epidemic hepatitis E virus (HEV) by means of cell culturing have been described previously. This paper reports the successful isolation of a sporadic HEV strain (G93-2) in human lung carcinoma cell (A549) cultures. The etiology, molecular and biological properties, and serological relationship of this new strain to other, epidemic HEV strains are described. The propagation of both sporadic and epidemic HEV strains in a cell culture system will facilitate vaccine research. PMID- 10473527 TI - A new sensitive serological assay for detection of lentivirus infections in small ruminants. AB - Lentivirus infections in small ruminants represent an economic problem affecting several European countries with important sheep-breeding industries. Programs for control and eradication of these infections are being initiated and require reliable screening assays. This communication describes the construction and evaluation of a new serological screening enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to maedi-visna virus (MVV) in sheep and to caprine arthritis encephalitis virus (CAEV) in goats. The solid phase is sensitized with a combination of the major core protein p25 of MVV produced in Escherichia coli and a peptide derived from the immunodominant region of the viral transmembrane protein gp46. The peptide carries an N-terminal biotin residue and is complexed with streptavidin prior to being coated. The new assay was evaluated with 2,336 sheep serum samples from different European countries with large differences in the levels of prevalence of MVV infections, and the results have been compared to those of the standard agar gel immunodiffusion test. Discrepant samples were analyzed by Western blotting with viral lysate, and most sera could be classified unambiguously. The estimated overall sensitivity of the new ELISA was 99.4% (95% confidence interval [CI], 98.4 to 99. 8%) and the specificity was 99.3% (95% CI, 98.7 to 99.6%). A limited set of goat sera (n = 212) was also analyzed, with similar results. These data indicate that the new assay is a reliable tool that can be used in control and eradication programs for small ruminant lentivirus infections. PMID- 10473528 TI - Evaluation of a commercially available immunoglobulin M capture enzyme-linked immunosorbent assay kit for diagnosing acute dengue infections. AB - Recently, commercially available kits for the detection of anti-dengue virus (anti-DEN) immunoglobulin M (IgM) antibodies have been developed. These standardized assays have greatly enhanced our ability to effectively diagnose DEN infections. We conducted an evaluation of a test kit manufactured by MRL Diagnostics Inc. that is designed to detect anti-DEN IgM antibodies. Eighty paired samples from DEN-infected individuals were tested by the MRL DEN Fever Virus IgM Capture enzyme-linked immunosorbent assay (ELISA), the PanBio Duo ELISA, the PanBio Rapid Immunochromatographic Test (PRIT), and the IgM-IgG antibody capture (MAC/GAC) ELISA. All infections were confirmed by either PCR assisted detection of DEN transcripts or by DEN isolation in C6/36 cells. Seventeen paired samples from individuals with no evidence of acute DEN infection were used as negative controls. The PRIT had the best sensitivity (100%), whereas the MAC/GAC ELISA and the PanBio Duo assay had the highest levels of specificity. The MRL ELISA and the PanBio Duo assay were the top performers when taking into consideration both sensitivity and specificity. All assays were able to detect DEN-specific antibodies in samples from patients with either primary or secondary infections, regardless of the infecting DEN serotype. PMID- 10473529 TI - Cloning and expression of an immunogenic membrane-associated protein of Helicobacter hepaticus for use in an enzyme-linked immunosorbent assay. AB - Helicobacter hepaticus is a bacterial pathogen that causes chronic active hepatitis and inflammatory bowel disease in mice. The purpose of this study was to develop a recombinant antigen-based enzyme-linked immunosorbent assay (ELISA) to detect H. hepaticus-infected mice. A genomic library of H. hepaticus was constructed and was screened with sera from H. hepaticus-infected mice. A 459-bp open reading frame that coded for an 18-kDa immunoreactive protein, MAP18, was identified. The gene had high identity with genes coding for outer membrane proteins of other bacteria, and the predicted amino acid sequence of MAP18 had a putative membrane-trafficking signal sequence and a putative signal peptidase II cleavage site. The recombinant protein was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein, GST-MAP18, and purified by affinity chromatography. The 44-kDa fusion protein was detected on Western blots probed with sera from H. hepaticus-infected mice but was not detected on blots probed with sera from mice infected with Helicobacter muridarum or Helicobacter bilis or with sera from mice free of Helicobacter infection. The GST-MAP18 fusion protein was used as an antigen in an ELISA to detect anti-H. hepaticus antibodies in sera from infected mice. This ELISA was compared to an H. hepaticus-specific ELISA that uses a detergent extract of H. hepaticus as the antigen. Sera from mice naturally and experimentally infected with H. hepaticus, H. bilis, or H. muridarum and sera from mice free of Helicobacter infection were evaluated. Both ELISAs performed with a high specificity (98%); however, the detergent extract based ELISA performed with a higher sensitivity (89%) than the recombinant protein-based ELISA (sensitivity, 66%). These data indicate that H. hepaticus carries a gene that encodes an immunogenic 18-kDa membrane-associated protein; however, antibodies to this protein are not detected in all infected mice. PMID- 10473530 TI - Th1 cytokine patterns in cervical human papillomavirus infection. AB - The host's immune response to cervical human papillomavirus (HPV) infection is poorly understood. In a longitudinal cohort of women with cervical HPV infections, defined by PCR-based HPV DNA testing, we used exfoliated cervical cells and reverse transcription-PCR to examine the cervical mucosal mRNA expression of cytokines involved in regulating cell-mediated immunity. We identified seven HPV-positive subjects who were found to have cleared their HPV infections 4 months later. In all seven, a T-helper type 1 (Th1) cytokine pattern (expression of gamma interferon and absence of interleukin-4) preceded clearance. The more variable cytokine patterns seen in HPV-negative subjects suggest that the Th1 pattern in the women with subsequent clearance was a response to the HPV infection. This contention is supported by additional cross-sectional data showing a Th1 pattern in a majority of HPV-positive women. This study establishes a feasible means for assessing local cytokine expression in the cervical milieu and demonstrates that a Th1 cytokine response is associated with subsequent clearance of cervical HPV infection. PMID- 10473531 TI - Detection of antibodies to Brucella cytoplasmic proteins in the cerebrospinal fluid of patients with neurobrucellosis. AB - The diagnosis of human neurobrucellosis usually relies on the detection of antibodies to Brucella lipopolysaccharide (LPS) in cerebrospinal fluid (CSF) by agglutination tests or enzyme-linked immunosorbent assay (ELISA). Here we describe the detection of immunoglobulin G (IgG) to cytoplasmic proteins (CP) of Brucella spp. by ELISA and Western blotting in seven CSF samples from five patients with neurobrucellosis. While IgG to CP (titers of 200 to 12, 800) and IgG to LPS (800 to 6,400) were found in the CSF of these patients, these antibodies were not detected in CSF samples from two patients who had systemic brucellosis without neurological involvement. The latter, however, had serum IgG and IgM to both LPS and CP. No reactivity to these antigens was found in CSF samples from 14 and 20 patients suffering from nonbrucellar meningitis and noninfectious diseases, respectively. These findings suggest that, in addition to its usefulness in the serological diagnosis of human systemic brucellosis, the ELISA with CP antigen can be used for the specific diagnosis of human neurobrucellosis. PMID- 10473532 TI - Identification of an IS711 element interrupting the wboA gene of Brucella abortus vaccine strain RB51 and a PCR assay to distinguish strain RB51 from other Brucella species and strains. AB - Brucella abortus vaccine strain RB51 is a natural stable attenuated rough mutant derived from the virulent strain 2308. The genetic mutations that are responsible for the roughness and the attenuation of strain RB51 have not been identified until now. Also, except for an assay based on pulsed-field gel electrophoresis, no other simple method to differentiate strain RB51 from its parent strain 2308 is available. In the present study, we demonstrate that the wboA gene encoding a glycosyltransferase, an enzyme essential for the synthesis of O antigen, is disrupted by an IS711 element in B. abortus vaccine strain RB51. Exploiting this feature, we developed a PCR assay that distinguishes strain RB51 from all other Brucella species and strains tested. PMID- 10473533 TI - Serological evidence of human infection with the protozoan Neospora caninum. AB - Neospora caninum is a protozoan parasite that is closely related to Toxoplasma gondii. Dogs are a definitive host. Prior to its discovery in 1988, N. caninum infection in animals was often mistakenly diagnosed as toxoplasmosis. Neosporosis in animals is characterized by encephalitis, abortion, and other conditions that clinically and pathologically resemble toxoplasmosis. The potential of N. caninum to infect humans is unknown. Therefore, evidence of human exposure to this parasite was sought by screening for antibodies in blood donors by indirect fluorescent antibody (IFA) tests and immunoblotting. Of 1,029 samples screened, 69 (6.7%) had titers of 1:100 by IFA testing. Fifty of the 69 (72%) sera that were positive for N. caninum were also negative for a closely related protozoan pathogen of humans, T. gondii. Immunoblot analysis confirmed the specificity of the positive sera for N. caninum antigens, with several sera recognizing multiple Neospora antigens with molecular masses similar to those of antigens recognized by monkey anti-N. caninum serum. An immunodominant antigen of approximately 35 kDa was observed with 12 sera. These data provide evidence of human exposure to N. caninum, although the antibody titers in healthy donors were low. The significance of human exposure to, and possible infection with, this parasite is unknown and warrants further study. PMID- 10473534 TI - Hepatitis B virus DNA in blood samples positive for antibodies to core antigen and negative for surface antigen. AB - Anti-hepatitis B core antigen (HBcAg)-positive hepatitis B surface antigen (HBsAg)-negative plasma samples from blood donors were tested by nested PCR. DNA positivity was more significantly associated with high levels of anti-HBcAg than with low levels of anti-HBsAg antibodies. Analysis of a dilution of anti-HBcAg antibodies might result in a more rational exclusion of anti-HBcAg-positive HBsAg negative samples, reducing the number of donations discarded and enabling more countries to incorporate anti-HBcAg testing. PMID- 10473536 TI - The calcineurin-dynamin 1 complex as a calcium sensor for synaptic vesicle endocytosis. AB - Exocytosis of synaptic vesicles is calcium-dependent, with synaptotagmin serving as the calcium sensor. Endocytosis of synaptic vesicles has also been postulated as a calcium-dependent process; however, an endocytic calcium sensor has not been found. We now report a physical association between the calcium-dependent phosphatase calcineurin and dynamin 1, a component of the synaptic endocytic machinery. The calcineurin-dynamin 1 interaction is calcium-dependent, with an EC(50) for calcium in the range of 0.1-0. 4 microM. Disruption of the calcineurin dynamin 1 interaction inhibits clathrin-mediated endocytosis. Thus, the calcium dependent formation of the calcineurin-dynamin 1 complex, delivered to the other endocytic coat proteins, provides a calcium-sensing mechanism that facilitates endocytosis. PMID- 10473535 TI - The oxidation of lipoproteins by monocytes-macrophages. Biochemical and biological mechanisms. PMID- 10473537 TI - Group V phospholipase A(2)-dependent induction of cyclooxygenase-2 in macrophages. AB - When exposed for prolonged periods of time (up to 20 h) to bacterial lipopolysaccharide (LPS) murine P388D(1) macrophages exhibit a delayed prostaglandin biosynthetic response that is entirely mediated by cyclooxygenase-2 (COX-2). Both the constitutive Group IV cytosolic phospholipase A(2) (cPLA(2)) and the inducible Group V secretory phospholipase A(2) (sPLA(2)) are involved in the cyclooxygenase-2-dependent generation of prostaglandins in response to LPS. Using the selective sPLA(2) inhibitor 3-(3-acetamide-1-benzyl-2-ethylindolyl-5 oxy)propane sulfonic acid (LY311727) and an antisense oligonucleotide specific for Group V sPLA(2), we found that induction of COX-2 expression is strikingly dependent on Group V sPLA(2), which was further confirmed by experiments in which exogenous Group V sPLA(2) was added to the cells. Exogenous Group V sPLA(2) was able to induce significant arachidonate mobilization on its own and to induce expression of the COX-2. None of these effects was observed if inactive Group V sPLA(2) was utilized, implying that enzyme activity is crucial for these effects to take place. Therefore, not only delayed prostaglandin production but also COX 2 gene induction are dependent on a catalytically active Group V sPLA(2). COX-2 expression was also found to be blunted by the Group IV cPLA(2) inhibitor methyl arachidonyl fluorophosphonate, which we have previously found to block Group V sPLA(2) induction as well. Collectively, the results support a model whereby Group IV cPLA(2) activation regulates the expression of Group V sPLA(2), which in turn is responsible for delayed prostaglandin production by regulating COX-2 expression. PMID- 10473538 TI - Molecular identification of the ryanodine receptor pore-forming segment. AB - A sequence motif, GXRXGGGXGD, located in the putative channel-forming domain, is conserved in all known ryanodine receptors and inositol 1,4,5-trisphosphate receptors. The functional significance of this conserved region was investigated by using site-directed mutagenesis together with functional assays consisting of Ca(2+) release measurements, [(3)H]ryanodine binding, and single channel recordings in planar lipid bilayers. We report here that single point mutations introduced into this region of the mouse cardiac ryanodine receptor reduce or abolish high affinity [(3)H]ryanodine binding. Single channel analysis revealed that a single substitution of alanine for glycine 4824 within this region reduced the single channel conductance by 97%, from 798 picosiemens (pS) for the wild type channel to 22 pS. The G4824A mutant channel was modulated by Ca(2+), Mg(2+), ATP, caffeine, ruthenium red, and ryanodine. Co-expression of the wild type and G4824A mutant proteins produced single channels that have intermediate unitary conductances of 516, 256, 176, and 60 pS. These data suggest that this conserved region constitutes an essential part of the ryanodine binding site and the channel conduction pathway of the ryanodine receptor. PMID- 10473539 TI - Abasic sites induce triplet-repeat expansion during DNA replication in vitro. AB - The occurrence of triplet-repeat expansion (TRE) during transmission of genetic information is involved in many neurological and neuromuscular diseases including Fragile X syndrome and myotonic dystrophy. DNA slippage during replicative synthesis appears to cause TRE. The causes of DNA slippage, however, remain mostly unknown. We investigated the effects of abasic sites on the occurrence of TRE during DNA replication in vitro using Escherichia coli Klenow polymerase I as the model polymerase. Here we show that a single abasic site analog, synthesized in the triplet-repeat tract at the 5' end of the template strand, induced dramatic TRE during DNA synthesis. The amount of TRE induced decreased when the abasic site was moved to the middle of the repeat tract, consistent with effectively decreasing the length of the repeat tract. Placing the abasic site in the primer did not induce TRE. TRE was sequence-dependent. The damage-induced increase in growing strand TRE depended on the sequence of the growing strand repeat as AAT approximately ATT > CAG > CTG. The expansions required replication from a primer complementary to the repeat tract. The expanded tracts were sequenced and contained multiple additions of the original repeat. The results imply that DNA damage can play a significant role in generating TRE in vivo. PMID- 10473540 TI - Differential rates of NTP hydrolysis by the mutant [S69G]RecA protein. Evidence for a coupling of NTP turnover to DNA strand exchange. AB - The x-ray crystal structure of the Escherichia coli RecA protein indicates that the phosphate groups of the nucleotide cofactor are bound by a loop whose amino acid sequence ((66)GPESSGKT(73)) corresponds to a consensus phosphate binding loop sequence (GXXXXGK[T/S]) found in many NTP-binding proteins. As part of an investigation of the role of the P-loop in ATP hydrolysis, we prepared a mutant RecA protein in which serine 69 was replaced by a glycine residue. We have found that the [S69G]RecA mutation has a differential effect on the hydrolysis of various nucleoside triphosphates. The [S69G]RecA protein catalyzes the single stranded DNA-dependent hydrolysis of rATP, ddATP, and dATP with turnover numbers of 10, 20, and 36 min(-1), respectively. The wild type RecA protein, in contrast, hydrolyzes each of these nucleoside triphosphates with similar turnover numbers of 20-24 min(-1). Significantly, the [S69G]RecA protein promotes strand exchange with all three nucleoside triphosphates, and the rate of strand exchange is directly proportional to the rate of hydrolysis of each of the nucleotide cofactors. These findings with the [S69G]RecA protein provide support for the existence of a mechanistic coupling between NTP hydrolysis and DNA strand exchange. PMID- 10473541 TI - Regulation of the miniature plasma membrane Ca(2+) channel I(min) by inositol 1,4,5-trisphosphate receptors. AB - I(min) is a plasma membrane-located, Ca(2+)-selective channel that is activated by store depletion and regulated by inositol 1,4, 5-trisphosphate (IP(3)). In the present work we examined the coupling between I(min) and IP(3) receptors in excised plasma membrane patches from A431 cells. I(min) was recorded in cell attached mode and the patches were excised into medium containing IP(3). In about 50% of experiments excision caused the loss of activation of I(min) by IP(3.) In the remaining patches activation of I(min) by IP(3) was lost upon extensive washes of the patch surface. The ability of IP(3) to activate I(min) was restored by treating the patches with rat cerebellar microsomes reach in IP(3) receptors but not by control forebrain microsomes. The re-activated I(min) had the same kinetic properties as I(min) when it is activated by Ca(2+)-mobilizing agonists in intact cells and by IP(3) in excised plasma membrane patches and it was inhibited by the I(crac) inhibitor SKF95365. We propose that I(min) is a form of I(crac) and is gated by IP(3) receptors. PMID- 10473542 TI - A new type of carbohydrate-deficient glycoprotein syndrome due to a decreased import of GDP-fucose into the golgi. AB - The fucosylation of glycoproteins was found to be deficient in a patient with a clinical phenotype resembling that of leukocyte adhesion deficiency type II (LAD II). While in LAD II hypofucosylation of glycoconjugates is secondary to an impaired synthesis of GDP-fucose due to a deficiency of the GDP-D-mannose-4, 6 dehydratase, synthesis of GDP-fucose was normal in our patient (Korner, C., Linnebank, M., Koch, H., Harms, E., von Figura, K., and Marquardt, T. (1999) J. Leukoc. Biol., in press). Import of GDP-fucose into Golgi-enriched vesicles was composed of a saturable, high affinity and a nonsaturable component. In our patient the saturable high affinity import of GDP-fucose was deficient, while import of UDP-galactose and the activity of GDPase, which generates the nucleoside phosphate required for antiport of GDP-fucose, were normal. Addition of L-fucose to the medium of fibroblasts restored the fucosylation of glycoproteins. We propose that this new form of carbohydrate-deficient glycoprotein syndrome is caused by impaired import of GDP-fucose into the Golgi. PMID- 10473543 TI - Reevaluation of the nucleotide cofactor specificity of the RecA protein from Bacillus subtilis. AB - The RecA protein from the Gram-positive bacterium, Bacillus subtilis, has been reported to catalyze dATP hydrolysis and to promote strand exchange in the presence of dATP but to have no ATP hydrolysis or ATP-dependent strand exchange activity (Lovett, C. M., Jr., and Roberts, J. W. (1985) J. Biol. Chem. 260, 3305 3313). The well characterized RecA protein from Escherichia coli, in contrast, catalyzes the hydrolysis of ATP and dATP at similar rates and can use either ATP or dATP as a cofactor for the strand exchange reaction. To explore this reported difference in nucleotide cofactor specificity in detail, we developed an overexpression system for the B. subtilis RecA protein and purified the protein to greater than 95% homogeneity. Contrary to the previous report, we find that the B. subtilis RecA protein catalyzes the hydrolysis of both dATP and ATP and can perform strand exchange using either dATP or ATP as a cofactor. Our results suggest that the inability of previous investigators to detect the ATP hydrolysis and ATP-dependent strand exchange activities of the B. subtilis RecA protein may have been due to the particular assay conditions that were used in the earlier study. PMID- 10473544 TI - Caspase inhibition by baculovirus P35 requires interaction between the reactive site loop and the beta-sheet core. AB - Baculovirus P35 is a universal substrate-inhibitor of the death caspases. Stoichiometric inhibition by P35 is correlated with cleavage of its reactive site loop (RSL) and formation of a stable P35.caspase complex through a novel but undefined mechanism. The P35 crystal structure predicts that the RSL associates with the beta-sheet core of P35 positioning the caspase cleavage site at the loop's apex. Here we demonstrate that proper interaction between the RSL and the beta-sheet core is critical for caspase inhibition, but not cleavage. Disruption of RSL interaction with the beta-sheet by substituting hydrophobic residues of the RSL's transverse helix alpha1 with destabilizing charged residues caused loss of caspase inhibition, without affecting P35 cleavage. Restabilization of the helix/sheet interaction by charge compensation from within the beta-sheet partially restored anti-caspase potency. Mutational effects on P35 helix/sheet interactions were confirmed by measuring intermolecular helix/sheet association with the yeast two-hybrid system. Moreover, the identification of P35 oligomers in baculovirus-infected cells suggested that similar P35 interactions occur in vivo. These findings indicate that P35's anti-caspase potency depends on a distinct conformation of the RSL which is required for events that promote stable, post-cleavage interactions and inhibition of the target caspase. PMID- 10473545 TI - Transport function and regulation of mitochondrial uncoupling proteins 2 and 3. AB - Uncoupling protein 1 (UCP1) dissipates energy and generates heat by catalyzing back-flux of protons into the mitochondrial matrix, probably by a fatty acid cycling mechanism. If the newly discovered UCP2 and UCP3 function similarly, they will enhance peripheral energy expenditure and are potential molecular targets for the treatment of obesity. We expressed UCP2 and UCP3 in Escherichia coli and reconstituted the detergent-extracted proteins into liposomes. Ion flux studies show that purified UCP2 and UCP3 behave identically to UCP1. They catalyze electrophoretic flux of protons and alkylsulfonates, and proton flux exhibits an obligatory requirement for fatty acids. Proton flux is inhibited by purine nucleotides but with much lower affinity than observed with UCP1. These findings are consistent with the hypothesis that UCP2 and UCP3 behave as uncoupling proteins in the cell. PMID- 10473546 TI - An archaebacterial ATPase, homologous to ATPases in the eukaryotic 26 S proteasome, activates protein breakdown by 20 S proteasomes. AB - In eukaryotes, the 20 S proteasome is the proteolytic core of the 26 S proteasome, which degrades ubiquitinated proteins in an ATP-dependent process. Archaebacteria lack ubiquitin and 26 S proteasomes but do contain 20 S proteasomes. Many archaebacteria, such as Methanococcus jannaschii, also contain a gene (S4) that is highly homologous to the six ATPases in the 19 S (PA700) component of the eukaryotic 26 S proteasome. To test if this putative ATPase may regulate proteasome function, we expressed it in Escherichia coli and purified the 50-kDa product as a 650-kDa complex with ATPase activity. When mixed with the well characterized 20 S proteasomes from Thermoplasma acidophilum and ATP, this complex stimulated degradation of several unfolded proteins 8-25-fold. It also stimulated proteolysis by 20 S proteasomes from another archaebacterium and mammals. This effect required ATP hydrolysis since ADP and the nonhydrolyzable analog, 5'-adenylyl beta, gamma-imidophosphate, were ineffective. CTP and to a lesser extent GTP and UTP were also hydrolyzed and also stimulated proteolysis. We therefore named this complex PAN for proteasome-activating nucleotidase. However, PAN did not promote the degradation of small peptides, which, unlike proteins, should readily diffuse into the proteasome. This ATPase complex appears to have been the evolutionary precursor of the eukaryotic 19 S complex, before the coupling of proteasome function to ubiquitination. PMID- 10473547 TI - Sequence, S-S bridges, and spectra of bovine transcobalamin expressed in Pichia pastoris. AB - Transcobalamin (TC) -encoding cDNA was isolated from a bovine mammary gland cDNA library. Hybridization of the cloned bovine TC-cDNA to RNA samples from bovine tissues showed that the most intensive synthesis of a TC positive 1.9-kilobase mRNA occurred in kidney, lymphatic nodes, and liver. Bovine TC was expressed in yeast Pichia pastoris, and the isolated recombinant protein showed cobalamin (Cbl) and receptor binding properties similar to TCs from other sources. Alignment of the related Cbl carriers (haptocorrins and intrinsic factors from other species) with bovine TC (414 residues) revealed four conservative clusters in the sequence (85-98, 137-147, 178-190, and 268-288), which may be responsible for Cbl binding. Three S-S bonds connected Cys residues 3-252, 98-294, and 147 190. Treatment with an S-S reducing agent caused liberation of Cbl from TC-Cbl. A significant change was observed in the TC-Cbl absorbance spectrum upon substitution of Co(2+)-coordinated H(2)O by azide. The reaction developed several orders of magnitude slower, and the spectral distortions were much stronger than those in free Cbl. This may be caused by significant deformation of the Cbl molecule and/or by its shielding when bound to TC. PMID- 10473548 TI - Structural and enzymatic properties of a gastric NADP(H)- dependent and retinal active alcohol dehydrogenase. AB - A class IV-type, gastric alcohol dehydrogenase (ADH) has been purified from frog (Rana perezi) tissues, meaning detection of this enzyme type also in nonmammalian vertebrates. However, the protein is unique among vertebrate ADHs thus far characterized in having preference for NADP(+) rather than NAD(+). Similarly, it deviates structurally from other class IV ADHs and has a phylogenetic tree position outside that of the conventional class IV cluster. The NADP(+) preference is structurally correlated with a replacement of Asp-223 of all other vertebrate ADHs with Gly-223, largely directing the coenzyme specificity. This residue replacement is expected metabolically to correlate with a change of the reaction direction catalyzed, from preferential alcohol oxidation to preferential aldehyde reduction. This is of importance in cellular growth regulation through retinoic acid formed from retinol/retinal precursors because the enzyme is highly efficient in retinal reduction (k(cat)/K(m) = 3.4.10(4) mM(-1) min(-1)). Remaining enzymatic details are also particular but resemble those of the human class I/class IV enzymes. However, overall structural relationships are distant (58-60% residue identity), and residues at substrate binding and coenzyme binding positions are fairly deviant, reflecting the formation of the new activity. The results are concluded to represent early events in the duplicatory origin of the class IV line or of a separate, class IV-type line. In both cases, the novel enzyme illustrates enzymogenesis of classes in the ADH system. The early origin (with tetrapods), the activity (with retinoids), and the specific location of this enzyme (gastric, like the gastric and epithelial location of the human class IV enzyme) suggest important functions of the class IV ADH type in vertebrates. PMID- 10473549 TI - Oxidative stress defense and deterioration of growth-arrested Escherichia coli cells. AB - Analysis of protein carbonylation demonstrates that the stasis-induced catalases and cytoplasmic superoxide dismutases (SOD) have a role in preventing accelerated protein oxidation during growth arrest of Escherichia coli cells. A larger number of proteins are carbonylated in cells lacking cytoplasmic SOD compared with cells lacking catalases, OxyR, or RpoS which, in turn, exhibit a larger number of oxidized proteins than the wild-type parent. Proteins exclusively oxidized during stasis in mutants lacking cytoplasmic SOD include GroEL, EF-G, and the acidic isoform of H-NS indicating that these mutants experience problems in peptide elongation and maintaining protein and DNA architecture. These mutants also survive stasis poorly. Likewise, but to a much lesser extent, mutations in oxyR, an oxidative stress regulator, shorten the life-span of stationary phase cells. The low plating efficiency of cells lacking OxyR is the result of their inability to grow on standard culture plates unless plating is performed anaerobically or with high concentration of catalase. In contrast, cells lacking cytoplasmic SOD appear to die prior to plating. Our data points to the importance of oxidative stress defense in stasis survival, and we also demonstrate that the life-span of growth-arrested wild-type E. coli cells can be significantly extended by omitting oxygen. PMID- 10473550 TI - Biological properties of human prolactin analogs depend not only on global hormone affinity, but also on the relative affinities of both receptor binding sites. AB - Zinc increases the affinity of human growth hormone (hGH) for the human prolactin receptor (hPRLR) due to the coordination of one zinc ion involving Glu-174(hGH) and His-18(hGH). In contrast, binding of hPRL to the hPRLR is zinc-independent. We engineered in binding site 1 of hPRL a hGH-like zinc coordination site, by mutating Asp-183(hPRL) (homologous to Glu-174(hGH)) into Glu (D183E mutation). This mutation was also introduced into G129R hPRL, a binding site 2 mutant (Goffin, V., Kinet, S., Ferrag, F., Binart, N., Martial, J. A. , and Kelly, P. A. (1996) J. Biol. Chem. 271, 16573-16579). These analogs were characterized using a stable clone expressing both the hPRLR and a PRLR-responsive reporter gene. The D183E mutation per se decreases the binding affinity and transcriptional activity of hPRL. However, this loss is partially rescued by the addition of zinc and the effect is much more marked on bioactivity than on binding affinity. These data indicate that the D183E mutation confers zinc sensitivity to hPRL biological properties. Due to an impaired site 2, the agonistic activity of G129R analog is almost nil. Although the double mutant D183E/G129R displays lower affinity ( approximately 1 log) compared with G129R hPRL, it unexpectedly recovers partial agonistic activity in the absence of zinc. Moreover, whereas zinc increases the affinity of D183E/G129R, it paradoxically abolishes its agonistic activity. Our results demonstrate that the biological properties of hPRL analogs do not necessarily parallel their overall affinity. Rather, the relative affinities of the individual binding sites 1 and 2 may play an even more important role. PMID- 10473551 TI - Cdc42 and Rac1 regulate the interaction of IQGAP1 with beta-catenin. AB - IQGAP1, a target of Cdc42 and Rac1 small GTPases, directly interacts with beta catenin and negatively regulates E-cadherin-mediated cell-cell adhesion by dissociating alpha-catenin from the cadherin-catenin complex in vivo (Kuroda, S., Fukata, M., Nakagawa, M., Fujii, K., Nakamura, T., Ookubo, T., Izawa, I., Nagase, T., Nomura, N., Tani, H., Shoji, I., Matsuura, Y., Yonehara, S., and Kaibuchi, K. (1998) Science 281, 832-835). Here we investigated how Cdc42 and Rac1 regulate the IQGAP1 function. IQGAP1 interacted with the amino-terminal region (amino acids 1-183) of beta-catenin, which contains the alpha-catenin-binding domain. IQGAP1 dissociated alpha-catenin from the beta-catenin-alpha-catenin complex in a dose-dependent manner in vitro. Guanosine 5'-(3-O-thio)triphosphate (GTPgammaS).glutathione S-transferase (GST)-Cdc42 and GTPgammaS. GST-Rac1 inhibited the binding of IQGAP1 to beta-catenin in a dose-dependent manner in vitro, whereas neither GDP.GST-Cdc42, GDP. GST-Rac1, nor GTPgammaS.GST-RhoA did. The coexpression of dominant active Cdc42 with IQGAP1 suppressed the dissociation of alpha-catenin from the cadherin-catenin complex induced by the overexpression of IQGAP1 in L cells expressing E-cadherin (EL cells). Consistent with this, the overexpression of either dominant negative Cdc42 or Rac1 resulted in the reduction of E-cadherin-mediated cell adhesive activity in EL cells. These results indicate that Cdc42 and Rac1 negatively regulate the IQGAP1 function by inhibiting the interaction of IQGAP1 with beta-catenin, leading to stabilization of the cadherin-catenin complex. PMID- 10473552 TI - Metronidazole resistance in the protozoan parasite Entamoeba histolytica is associated with increased expression of iron-containing superoxide dismutase and peroxiredoxin and decreased expression of ferredoxin 1 and flavin reductase. AB - To obtain insight into the mechanism of metronidazole resistance in the protozoan parasite Entamoeba histolytica, amoeba trophozoites were selected in vitro by stepwise exposures to increasing amounts of metronidazole, starting with sublethal doses of 4 microM. Subsequently, amoebae made resistant were able to continuously multiply in the presence of a 40 microM concentration of the drug. In contrast to mechanisms of metronidazole resistance in other protozoan parasites, resistant amoebae did not substantially down-regulate pyruvate:ferredoxin oxidoreductase or up-regulate P-glycoproteins, but exhibited increased expression of iron-containing superoxide dismutase (Fe-SOD) and peroxiredoxin and decreased expression of flavin reductase and ferredoxin 1. Episomal transfection and overexpression of the various antioxidant enzymes revealed significant reduction in susceptibility to metronidazole only in those cells overexpressing Fe-SOD. Reduction was highest in transfected cells simultaneously overexpressing Fe-SOD and peroxiredoxin. Although induced overexpression of Fe-SOD did not confer metronidazole resistance to the extent found in drug-selected cells, transfected cells quickly adapted to constant exposures of otherwise lethal metronidazole concentrations. Moreover, metronidazole selection of transfected amoebae favored retention of the Fe-SOD containing plasmid. These results strongly suggest that peroxiredoxin and, in particular, Fe-SOD together with ferredoxin 1 are important components involved in the mechanism of metronidazole resistance in E. histolytica. PMID- 10473553 TI - Substrate- and inhibitor-induced conformational changes in the yeast V-ATPase provide evidence for communication between the catalytic and proton-translocating sectors. AB - The vacuolar-type H(+)-ATPases (V-ATPases) are composed of two distinct sectors, a catalytic complex (V(1)) involved in ATP hydrolysis and a membrane-associated complex (V(0)) mediating proton translocation across a lipid bilayer. To date, little is known about the mechanism by which these two functions are coupled. We sought to examine the impact of nucleotide and cation binding on the structure of the core components of the catalytic complex and to determine whether conformational changes within the catalytic complex impact subunits of the membrane-associated complex. Nucleotide- and cation- induced changes in the catalytic core of the V-ATPase were investigated by monitoring changes in the rate and pattern of tryptic digests. ATP.Mg-induced changes were detected in both the catalytic (Vma1p or 69 kDa) and the regulatory subunits (Vma2p or 60 kDa) of the V(1) sector. ATP alone increased the rate of trypsinization of the regulatory subunit, but did not have any effect on Vma1p. Surprisingly, ATP also had an impact on the 95-kDa subunit, a component of the V(0) sector of the V-ATPase. Although the presence of divalent cations had no impact on the V(1) sector, the rate of trypsinization of the 95-kDa subunit was greatly enhanced. The effect of divalent cations on the structure of the 95-kDa subunit was abrogated when trypsinization was performed in the absence of the catalytic sector. Addition of bafilomycin A(1), a V-ATPase inhibitor that putatively binds to the 95-kDa subunit, increased the rate of trypsinization of the catalytic subunit. These data suggest that structural alterations within the V(1) sector result in alterations within the V(0) sector and vice versa. Clearly, a structural link must exist to couple the two sectors. The 95-kDa subunit is ideally suited to fulfill this role. Hydropathy analysis suggests a bipartite structure, with the NH(2)-terminal portion predicted to lie in an aqueous environment and the C terminal portion predicted to contain 6 transmembrane segments. Tryptic digests of sealed vacuolar vesicles and immunofluorescence studies revealed that the large hydrophilic NH(2)-terminal domain of the 95-kDa subunit is localized toward the cytosol. This region therefore is ideally positioned to interact with components of the V(1) complex, potentially functioning as the elusive link between the two sectors of the V-ATPase. PMID- 10473554 TI - Energetics and topology of CzcA, a cation/proton antiporter of the resistance nodulation-cell division protein family. AB - The membrane-bound CzcA protein, a member of the resistance-nodulation-cell division (RND) permease superfamily, is part of the CzcCB(2)A complex that mediates heavy metal resistance in Ralstonia sp. CH34 by an active cation efflux mechanism driven by cation/proton antiport. CzcA was purified to homogeneity after expression in Escherichia coli, reconstituted into proteoliposomes, and the kinetics of heavy metal transport by CzcA was determined. CzcA is composed of 12 transmembrane alpha-helices and two large periplasmic domains. Two conserved aspartate and a glutamate residue in one of these transmembrane spans are essential for heavy metal resistance and proton/cation antiport but not for facilitated diffusion of cations. Generalization of the resulting model for the function of CzcA as a two-channel pump might help to explain the functions of other RND proteins in bacteria and eukaryotes. PMID- 10473555 TI - Nrf2, a Cap'n'Collar transcription factor, regulates induction of the heme oxygenase-1 gene. AB - Stress response elements, which mediate induction of the mouse heme oxygenase-1 (HO-1) gene by several agents, resemble the binding site for the activator protein-1 (Jun/Fos), Maf, and Cap'n'Collar/basic leucine zipper (CNC-bZIP) families of proteins. In L929 fibroblasts, significant activation of an HO-1 enhancer-reporter fusion gene was observed only with the CNC-bZIP class of proteins with Nrf2 exhibiting the highest level of trans-activation, between 25- and 30-fold. To further examine the role of this factor in HO-1 gene regulation, a dominant-negative mutant, Nrf2M, was generated and conditionally expressed in L929 cells. The mutant protein was detected in cytoplasmic and nuclear fractions but did not affect cell growth. Under conditions of Nrf2M overexpression, HO-1 mRNA accumulation in response to heme, cadmium, zinc, arsenite, and tert butylhydroquinone was inhibited by 85-95%. In contrast, overexpression of a dominant-negative mutant of c-Jun decreased L929 cell growth but did not inhibit HO-1 gene activation. Nrf2 does not homodimerize, but CNC-bZIP.small Maf protein heterodimers and Nrf2. Jun protein complexes are proposed to function as trans activators. Co-expression of Jun proteins or p18, however, had no significant affect or inhibited Nrf2-mediated trans-activation. Taken together, these results implicate Nrf2 in the induction of the HO-1 gene but suggest that the Nrf2 partner in this function is a factor other than p18 or Jun proteins. PMID- 10473556 TI - Involvement of the amino terminus of the B(2) receptor in agonist-induced receptor dimerization. AB - The mechanisms and the functional importance of G-protein-coupled receptor dimerization are poorly understood. We therefore analyzed dimerization of the bradykinin B(2) receptor. The binding of the agonist bradykinin to the B(2) receptor endogenously expressed on PC-12 cells led to the formation of receptor dimers, whereas the B(2) antagonist HOE140 did not induce dimerization, suggesting that B(2) receptor dimerization was linked to receptor activation. Addition of a peptide corresponding to the amino terminus of the receptor reduced the amount of detected B(2) receptor dimers, whereas peptides derived from the extracellular loops had no effect. To further analyze the role of the amino terminus of the receptor in receptor dimerization, we created two different rat B(2) receptor variants with truncated amino termini, B(2)(53) and B(2)(65), starting at amino acids 53 and 65. In contrast to the wild-type B(2) receptor and to B(2)(53), bradykinin did not induce dimerization of the B(2)(65) receptor. Both receptor variants were similar to the wild-type B(2) receptor with respect to agonist binding and signal generation. However, B(2)(65) was not phosphorylated, did not desensitize, and was not downregulated upon bradykinin stimulation. Likewise, antibodies directed to the amino terminus of the receptor partially reduced internalization of [(3)H]bradykinin on PC-12 cells. These findings suggest that the amino terminus of the B(2) receptor is necessary for triggering agonist-induced B(2) receptor dimerization, and receptor dimers are involved in receptor-mediated signal attenuation. PMID- 10473557 TI - A 12-amino acid stretch in the hypervariable region of the spike protein S1 subunit is critical for cell fusion activity of mouse hepatitis virus. AB - The spike (S) glycoprotein of mouse hepatitis virus (MHV) plays a major role in the viral pathogenesis. It is often processed into the N-terminal S1 and the C terminal S2 subunits that were evidently important for binding to cell receptor and inducing cell-cell fusion, respectively. As a consequence of cell-cell fusion, most of the naturally occurring infections of MHV are associated with syncytia formation. So far, only MHV-2 was identified to be fusion-negative. In this study, the S gene of MHV-2 was molecularly cloned, and the nucleotide sequence was determined. The MHV-2 S protein lacks a 12-amino acid stretch in the S1 hypervariable region from amino acid residue 446 to 457 when compared with the fusion-positive strain MHV-JHM. In addition, there are three amino acid substitutions in the S2 subunit, Tyr-1144 to Asp, Glu-1165 to Asp, and Arg-1209 to Lys. The cloned MHV-2 S protein exhibited the fusion-negative property in DBT cells as the intrinsic viral protein. Furthermore, similar to the fusion-positive MHV-JHM strain, proteolytic cleavage activity was detected both in DBT cells infected with the fusion-negative MHV-2 and in the transfected cells that expressed the cloned MHV-2 S protein. Domain swapping experiments demonstrated that the 12-amino acid stretch missing in the MHV-2 S1 subunit, but not the proteolytic cleavage site, was critical for the cell-fusion activity of MHV. PMID- 10473558 TI - The role of individual SH2 domains in mediating association of phospholipase C gamma1 with the activated EGF receptor. AB - The two SH2 (Src homology domain 2) domains present in phospholipase C-gamma1 (PLC-gamma1) were assayed for their capacities to recognize the five autophosphorylation sites in the epidermal growth factor receptor. Plasmon resonance and immunological techniques were employed to measure interactions between SH2 fusion proteins and phosphotyrosine-containing peptides. The N-SH2 domain recognized peptides in the order of pY1173 > pY992 > pY1068 > pY1148 >> pY1086, while the C-SH2 domain recognized peptides in the order of pY992 > pY1068 > pY1148 >> pY1086 and pY1173. The major autophosphorylation site, pY1173, was recognized only by the N-SH2 domain. Contributions of the N-SH2 and C-SH2 domains to the association of the intact PLC-gamma1 molecule with the activated epidermal growth factor (EGF) receptor were assessed in vivo. Loss of function mutants of each SH2 domain were produced in a full-length epitope-tagged PLC-gamma1. After expression of the mutants, cells were treated with EGF and association of exogenous PLC-gamma1 with EGF receptors was measured. In this context the N-SH2 is the primary contributor to PLC-gamma1 association with the EGF receptor. The combined results suggest an association mechanism involving the N-SH2 domain and the pY1173 autophosphorylation site as a primary event and the C-SH2 domain and the pY992 autophosphorylation site as a secondary event. PMID- 10473559 TI - A noninvasive fluorimetric procedure for measurement of membrane potential. Quantification of the NADPH oxidase-induced depolarization in activated neutrophils. AB - The electrogenic activity of the NADPH oxidase is associated with depolarization of the plasma membrane in activated neutrophils. The magnitude and consequences of this depolarization, however, remain unknown. Neutrophils are not amenable to electrophysiological determinations of membrane potential by current clamp. Instead, the occurrence of depolarization has been inferred from the use of potential-sensitive fluorescent dyes. However, such dyes partition into intracellular organelles and may yield erroneous results, particularly because the NADPH oxidase resides largely in secretory granules, where it has been claimed to become activated. We confirmed the intracellular generation of oxidase products using dihydrorhodamine, which is converted to the fluorescent rhodamine 123 when oxidized. Rhodamine 123 accumulated inside endomembrane organelles in both neutrophils and in differentiated HL60 cells, where it co-localized with the primary granule marker CD63. To estimate the surface membrane potential without interference from organelles, we devised a method based on the voltage-driven uptake of Mn(2+) across the plasmalemma. The uptake of Mn(2+) through calcium release-activated channels was measured as the rate of Indo-1 fluorescence quenching in thapsigargin-treated cells. The rate of Mn(2+) influx was found to vary when the membrane potential was manipulated using conductive ionophores and also when the NADPH oxidase was activated. A calibration curve in the positive potential range was constructed using the Na(+) ionophore SQI-Pr. Using this calibration, the membrane potential of phorbol ester-activated neutrophils was found to reach +58 +/- 6 mV, a sustained depolarization of over 100 mV compared with the resting potential. The depolarization was greatly diminished when the NADPH oxidase was inhibited with diphenylene iodonium. Together, these results indicate that the NADPH oxidase can generate a large depolarization of the plasmalemma, which should suffice to activate a variety of voltage-gated channels, including the outwardly rectifying H(+) conductance. PMID- 10473560 TI - Use of an antisense strategy to dissect the signaling role of protein-tyrosine phosphatase alpha. AB - The protein-tyrosine phosphatase PTPalpha has been proposed to play an important role in controlling the dephosphorylation of a number of key signaling proteins and in regulating insulin signaling. To examine the potential cellular functions and physiological substrates of PTPalpha, a potent phosphorothioate oligonucleotide-based antisense strategy was developed that specifically depleted endogenous PTPalpha from 3T3-L1 adipocytes. The antisense probe, alphaAS1, achieved PTPalpha depletion levels normally of >/=85% and which varied up to levels where PTPalpha was not detected at all. Elimination of PTPalpha by 85% inhibited c-Src activity by 80%. Abolishing PTPalpha to levels undetected did not alter the tyrosine dephosphorylation of the insulin receptor or insulin receptor substrate proteins. Moreover, the ability of insulin to activate ERK2 or to stimulate DNA synthesis was not altered by alphaAS1. It is concluded that endogenous PTPalpha is a key regulator of c-Src activity in 3T3-L1 adipocytes and that PTPalpha is not required for the dephosphorylation of the insulin receptor or the insulin receptor substrate proteins or for the regulation of several downstream insulin signaling events in 3T3-L1 adipocytes. Finally, the development of the antisense probe, alphaAS1, provides an important molecular tool of general applicability for further dissecting the roles and precise targets of endogenous PTPalpha. PMID- 10473561 TI - Chimeric analysis of a neuronal nicotinic acetylcholine receptor reveals amino acids conferring sensitivity to alpha-bungarotoxin. AB - We have investigated the molecular determinants responsible for alpha bungarotoxin (alphaBgtx) binding to nicotinic acetylcholine receptors through chimeric analysis of two homologous alpha subunits, one highly sensitive to alphaBgtx block (alpha1) and the other, alphaBgtx-insensitive (alpha3). By replacing rat alpha3 residues 184-191 with the corresponding region from the Torpedo alpha1 subunit, we introduced a cluster of five alpha1 residues (Trp-184, Trp-187, Val-188, Tyr-189, and Thr-191) into the alpha3 subunit. Functional activity and alphaBgtx sensitivity were assessed following co-expression in Xenopus oocytes of the chimeric alpha3 subunit (alpha3/alpha1[5]) with either rat beta2 or beta4 subunits. Agonist-evoked responses of alpha3/alpha1[5]-containing receptors were blocked by alphaBgtx with nanomolar affinity (IC(50) values: 41 nM for alpha3/alpha1[5]beta2 and 19 nM for alpha3/alpha1[5]beta4). Furthermore, receptors containing the single point mutation alpha3K189Y acquire significant sensitivity to alphaBgtx block (IC(50) values: 186 nM for alpha3K189Ybeta2 and 179 nM for alpha3K189Ybeta4). Another alpha3 chimeric subunit, alpha3/alpha7[6], similar to alpha3/alpha1[5] but incorporating the corresponding residues from the alphaBgtx-sensitive alpha7 subunit, also conferred potent alphaBgtx sensitivity to chimeric receptors when co-expressed with the beta4 subunit (IC(50) value = 31 nM). Our findings demonstrate that the residues between positions 184 and 191 of the alphaBgtx-sensitive subunits alpha1 and alpha7 play a critical functional role in the interaction of alphaBgtx with nicotinic acetylcholine receptors sensitive to this toxin. PMID- 10473562 TI - Ca(2+) inactivation sites are located in the COOH-terminal quarter of recombinant rabbit skeletal muscle Ca(2+) release channels (ryanodine receptors). AB - Ca(2+) activation of skeletal (RyR1) and cardiac (RyR2) muscle Ca(2+) release channels (ryanodine receptors) occurs with EC(50) values of about 1 microM. Ca(2+) inactivation occurs with an IC(50) value of about 3.7 mM for RyR1, but RyR2 shows little inactivation, even at >100 mM Ca(2+). In an attempt to localize the low affinity Ca(2+) binding sites responsible for Ca(2+) inactivation in RyR1, chimeric RyR1/RyR2 molecules were constructed. Because [(3)H]ryanodine binds only to open channels, and because channel opening and closing are Ca(2+) dependent, the Ca(2+) dependence of [(3)H]ryanodine binding was used as an indirect measurement of Ca(2+) release channel opening and closing. IC(50) values for [(3)H]ryanodine binding suggested that Ca(2+) affinity for the low affinity Ca(2+) inactivation sites was unchanged in a chimera in which a glutamate-rich sequence (amino acids 1743-1964) in RyR1 was replaced with the corresponding, less acidic sequence from RyR2. Ca(2+) affinity (IC(50)) for low affinity Ca(2+) inactivation sites was intermediate in RyR1/RyR2 chimeras containing RyR2 amino acids 3726-4186 (RF9), 4187-4628 (RF10), or 4629-5037 (RF11), was closer to RyR2 values in RyR1 chimeras with longer RyR2 replacements (RF9/10 or RF10/11), and was indistinguishable from RyR2 in RyR1 containing all three RyR2 replacements (RF9/10/11). These data suggest that multiple low affinity Ca(2+) binding sites or multiple components of a low affinity Ca(2+) binding site are located between amino acids 3726 and 5037 and that their effects on Ca(2+) inactivation of the release channel are cooperative. Measurement of Ca(2+) activation of [(3)H]ryanodine binding showed that chimeras RF10, RF9/10, and RF9/10/11 were more sensitive to Ca(2+) than was either RyR1 or RyR2. Measurement of caffeine activation of Ca(2+) release in vivo showed that chimeras RF9, RF10, RF9/10, RF10/11, and RF9/10/11 were more sensitive to caffeine than wild-type RyR1. These results suggest that Ca(2+) and caffeine activation sites also involve COOH terminal sequences in RyR1 and RyR2. PMID- 10473563 TI - Phospholipase C-delta1 is activated by capacitative calcium entry that follows phospholipase C-beta activation upon bradykinin stimulation. AB - To characterize the regulatory mechanism of phospholipase C-delta1 (PLC-delta1) in the bradykinin (BK) receptor-mediated signaling pathway, we used a clone of PC12 cells, which stably overexpress PLC-delta1 (PC12-D1). Stimulation with BK induced a significantly higher Ca(2+) elevation and inositol 1,4,5-trisphosphate (IP(3)) production with a much lower half-maximal effective concentration (EC(50)) of BK in PC12-D1 cells than in wild type (PC12-W) or vector-transfected (PC12-V) cells. However, BK-induced intracellular Ca(2+) release and IP(3) generation was similar between PC12-V and PC12-D1 cells in the absence of extracellular Ca(2+), suggesting that the availability of extracellular Ca(2+) is essential to the activation of PLC-delta1. When PC12-D1 cells were treated with agents that induce Ca(2+) influx, more IP(3) was produced, suggesting that the Ca(2+) entry induces IP(3) production in PC12-D1 cells. Furthermore, the additional IP(3) production after BK-induced capacitative calcium entry was detected in PC12-D1 cells, suggesting that PLC-delta1 is mainly activated by capacitative calcium entry. When cells were stimulated with BK in the presence of extracellular Ca(2+), [(3)H]norepinephrine secretion was much greater from PC12 D1 cells than from PC12-V cells. Our results suggest that PLC-delta1 is activated by capacitative calcium entry following the activation of PLC-beta, additively inducing IP(3) production and Ca(2+) rise in BK-stimulated PC12 cells. PMID- 10473564 TI - Human ceruloplasmin. Intramolecular electron transfer kinetics and equilibration. AB - Pulse radiolytic reduction of disulfide bridges in ceruloplasmin yielding RSSR(-) radicals induces a cascade of intramolecular electron transfer (ET) processes. Based on the three-dimensional structure of ceruloplasmin identification of individual kinetically active disulfide groups and type 1 (T1) copper centers, the following is proposed. The first T1 copper(II) ion to be reduced in ceruloplasmin is the blue copper center of domain 6 (T1A) by ET from RSSR(-) of domain 5. The rate constant is 28 +/- 2 s(-1) at 279 K and pH 7.0. T1A is in close covalent contact with the type 3 copper pair and indeed electron equilibration between T1A and the trinuclear copper center in the domain 1-6 interface takes place with a rate constant of 2.9 +/- 0.6 s(-1). The equilibrium constant is 0.17. Following reduction of T1A Cu(II), another ET process takes place between RSSR(-) and T1B copper(II) of domain 4 with a rate constant of 3.9 +/- 0.8. No reoxidation of T1B Cu(I) could be resolved. It appears that the third T1 center (T1C of domain 2) is not participating in intramolecular ET, as it seems to be in a reduced state in the resting enzyme. PMID- 10473565 TI - Human endothelial cell life extension by telomerase expression. AB - Normal human endothelial cells, like other somatic cells in culture, divide a limited number of times before entering a nondividing state called replicative senescence. Expression of the catalytic component of human telomerase, human telomerase reverse transcriptase (hTERT), extends the life span of human fibroblasts and retinal pigment epithelial cells beyond senescence without causing neoplastic transformation (Bodnar, A. G., Ouellette, M., Frolkis, M., Holt, S. E., Chiu, C. P., Morin, G. B., Harley, C. B., Shay, J. W., Lichtsteiner, S., and Wright, W. E. (1998) Science 279, 349-352; Jiang, X., Jimenez, G., Chang, E., Frolkis, M., Kusler, B., Sage, M., Beeche, M., Bodnar, A., Wahl, G., Tlsty, T., and Chiu, C.-P. (1999) Nat. Genet. 21, 111-114). Here, we show that both human large vessel and microvascular endothelial cells also bypass replicative senescence after introduction of hTERT. For the first time, we report that hTERT expression in these life-extended vascular cells does not affect their differentiated and functional phenotype and that these cells maintain their angiogenic potential in vitro. Furthermore, hTERT(+) microvascular endothelial cells have normal karyotype, and hTERT(+) endothelial cell strains do not exhibit a transformed phenotype. Relative to parental cells at senescence, hTERT expressing endothelial cells exhibit resistance to induction of apoptosis by a variety of different conditions. Such characteristics are highly desirable for designing vascular transplantation and gene therapy delivery systems in vivo. PMID- 10473566 TI - Oligopeptidase B from Trypanosoma brucei, a new member of an emerging subgroup of serine oligopeptidases. AB - Trypanosoma brucei contains a soluble serine oligopeptidase (OP-Tb) that is released into the host bloodstream during infection, where it has been postulated to participate in the pathogenesis of African trypanosomiasis. Here, we report the identification of a single copy gene encoding the T. brucei oligopeptidase and a homologue from the related trypanosomatid pathogen Leishmania major. The enzymes encoded by these genes belong to an emerging subgroup of the prolyl oligopeptidase family of serine hydrolases, referred to as oligopeptidase B. The trypanosomatid oligopeptidases share 70% amino acid sequence identity with oligopeptidase B from the intracellular pathogen Trypanosoma cruzi, which has a demonstrated role in mammalian host cell signaling and invasion. OP-Tb exhibited no activity toward the prolyl oligopeptidase substrate H-Gly-Pro-7-amido-4 methylcoumarin. Instead, it had activity toward substrates of trypsin-like enzymes, particularly those that have basic amino acids in both P(1) and P(2) (e.g. benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin k(cat)/K(m) = 529 s(-1) microM(-1)). The activity of OP-Tb was enhanced by reducing agents and by polyamines, suggesting that these agents may act as in vivo regulators of OP-Tb activity. This study provides the basis of the characterization of a novel subgroup of serine oligopeptidases from kinetoplastid protozoa with potential roles in pathogenesis. PMID- 10473567 TI - An Escherichia coli mutant quinol:fumarate reductase contains an EPR-detectable semiquinone stabilized at the proximal quinone-binding site. AB - The EPR and thermodynamic properties of semiquinone (SQ) species stabilized by mammalian succinate:quinone reductase (SQR) in situ in the mitochondrial membrane and in the isolated enzyme have been well documented. The equivalent semiquinones in bacterial membranes have not yet been characterized, either in SQR or quinol:fumarate reductase (QFR) in situ. In this work, we describe an EPR detectable QFR semiquinone using Escherichia coli mutant QFR (FrdC E29L) and the wild-type enzyme. The SQ exhibits a g = 2.005 signal with a peak-to-peak line width of approximately 1.1 milliteslas at 150 K, has a midpoint potential (E(m(pH 7.2))) of -56.6 mV, and has a stability constant of approximately 1.2 x 10(-2) at pH 7.2. It shows extremely fast spin relaxation behavior with a P(1/2) value of >>500 milliwatts at 150 K, which closely resembles the previously described SQ species (SQ(s)) in mitochondrial SQR. This SQ species seems to be present also in wild-type QFR, but its stability constant is much lower, and its signal intensity is near the EPR detection limit around neutral pH. In contrast to mammalian SQR, the membrane anchor of E. coli QFR lacks heme; thus, this prosthetic group can be excluded as a spin relaxation enhancer. The trinuclear iron-sulfur cluster FR3 in the [3Fe-4S](1+) state is suggested as the dominant spin relaxation enhancer of the SQ(FR) spins in this enzyme. E. coli QFR activity and the fast relaxing SQ species observed in the mutant enzyme are sensitive to the inhibitor 2-n-heptyl-4 hydroxyquinoline N-oxide (HQNO). In wild-type E. coli QFR, HQNO causes EPR spectral line shape perturbations of the iron-sulfur cluster FR3. Similar spectral line shape changes of FR3 are caused by the FrdC E29L mutation, without addition of HQNO. This indicates that the SQ and the inhibitor-binding sites are located in close proximity to the trinuclear iron-sulfur cluster FR3. The data further suggest that this site corresponds to the proximal quinone-binding site in E. coli QFR. PMID- 10473568 TI - Cloning and expression of a proteoglycan UDP-galactose:beta-xylose beta1,4 galactosyltransferase I. A seventh member of the human beta4 galactosyltransferase gene family. AB - A seventh member of the human beta4-galactosyltransferase family, beta4Gal-T7, was identified by BLAST analysis of expressed sequence tags. The coding region of beta4Gal-T7 depicts a type II transmembrane protein with sequence similarity to beta4-galactosyltransferases, but the sequence was distinct in known motifs and did not contain the cysteine residues conserved in the other six members of the beta4Gal-T family. The genomic organization of beta4Gal-T7 was different from previous beta4Gal-Ts. Expression of beta4Gal-T7 in insect cells showed that the gene product had beta1,4-galactosyltransferase activity with beta-xylosides, and the linkage formed was Galbeta1-4Xyl. Thus, beta4Gal-T7 represents galactosyltransferase I enzyme (xylosylprotein beta1, 4-galactosyltransferase; EC 2.4.1.133), which attaches the first galactose in the proteoglycan linkage region GlcAbeta1-3Galbeta1-3Galbeta1-4Xylbeta1-O-Ser. Sequence analysis of beta4Gal-T7 from a fibroblast cell line of a patient with a progeroid syndrome and signs of the Ehlers-Danlos syndrome, previously shown to exhibit reduced galactosyltransferase I activity (Quentin, E., Gladen, A., Roden, L., and Kresse, H. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 1342-1346), revealed two inherited allelic variants, beta4Gal-T7(186D) and beta4Gal-T7(206P), each with a single missense substitution in the putative catalytic domain of the enzyme. beta4Gal T7(186D) exhibited a 4-fold elevated K(m) for the donor substrate, whereas essentially no activity was demonstrated with beta4Gal-T7(206P). Molecular cloning of beta4Gal-T7 should facilitate general studies of its pathogenic role in progeroid syndromes and connective tissue disorders with affected proteoglycan biosynthesis. PMID- 10473569 TI - Horseshoe crab hemocyte-derived antimicrobial polypeptides, tachystatins, with sequence similarity to spider neurotoxins. AB - Antimicrobial peptides, named tachystatins A, B, and C, were identified from hemocytes of the horseshoe crab Tachypleus tridentatus. Tachystatins exhibited a broad spectrum of antimicrobial activity against Gram-negative and Gram-positive bacteria and fungi. Of these tachystatins, tachystatin C was most effective. Tachystatin A is homologous to tachystatin B, but tachystatin C has no significant sequence similarity to tachystatins A and B. Tachystatins A and B showed sequence similarity to omega-agatoxin-IVA of funnel web spider venom, a potent blocker of voltage-dependent calcium channels. However, they exhibited no blocking activity of the P-type calcium channel in rat Purkinje cells. Tachystatin C also showed sequence similarity to several insecticidal neurotoxins of spider venoms. Tachystatins A, B, and C bound significantly to chitin. A causal relationship was observed between chitin binding activity and antifungal activity. Tachystatins caused morphological changes against a budding yeast, and tachystatin C had a strong cell lysis activity. The septum between mother cell and bud, a chitin-rich region, was stained by fluorescence-labeled tachystatin C, suggesting that the primary recognizing substance on the cell wall is chitin. As horseshoe crab is a close relative of the spider, tachystatins and spider neurotoxins may have evolved from a common ancestral peptide, with adaptive functions. PMID- 10473570 TI - Bacillus subtilis contains two small c-type cytochromes with homologous heme domains but different types of membrane anchors. AB - We demonstrate that the cccB gene, identified in the Bacillus subtilis genome sequence project, is the structural gene for a 10-kDa membrane-bound cytochrome c(551) lipoprotein described for the first time in B. subtilis. Apparently, CccB corresponds to cytochrome c(551) of the thermophilic bacterium Bacillus PS3. The heme domain of B. subtilis cytochrome c(551) is very similar to that of cytochrome c(550), a protein encoded by the cccA gene and anchored to the membrane by a single transmembrane polypeptide segment. Thus, B. subtilis contains two small, very similar, c-type cytochromes with different types of membrane anchors. The cccB gene is cotranscribed with the yvjA gene, and transcription is repressed by glucose. Mutants deleted for cccB or yvjA-cccB show no apparent growth, sporulation, or germination defect. YvjA is not required for the synthesis of cytochrome c(551), and its function remains unknown. PMID- 10473571 TI - Enzyme I(Ntr) from Escherichia coli. A novel enzyme of the phosphoenolpyruvate dependent phosphotransferase system exhibiting strict specificity for its phosphoryl acceptor, NPr. AB - The phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) phosphorylates sugars and regulates cellular metabolic processes using a phosphoryl transfer chain including the general energy coupling proteins, Enzyme I (EI) and HPr as well as the sugar-specific Enzyme II complexes. Analysis of the Escherichia coli genome has revealed the presence of 5 paralogues of EI and 5 paralogues of HPr, most of unknown function. The ptsP gene encodes an EI paralogue designated Enzyme I(nitrogen) (EI(Ntr)), and two genes located in the rpoN operon encode PTS protein paralogues, NPr and IIA(Ntr), both implicated in the regulation of sigma(54) activity. The ptsP gene was polymerase chain reaction amplified from the E. coli chromosome and cloned into an overexpression vector allowing the overproduction and purification of EI(Ntr). EI(Ntr) was shown to phosphorylate NPr in vitro using either a [(32)P]PEP-dependent protein phosphorylation assay or a quantitative sugar phosphorylation assay. EI(Ntr) phosphorylated NPr but not HPr, whereas Enzyme I exhibited a strong preference for HPr. These two pairs of proteins (EI(Ntr)/NPr and EI/HPr) thus exhibit little cross-reactivity. Phosphoryl transfer from PEP to NPr catalyzed by EI(Ntr) has a pH optimum of 8.0, is dependent on Mg(2+), is stimulated by high ionic strength, and exhibits two K(m) values for NPr (2 and 10 microM) possibly because of negative cooperativity. The results suggest that E. coli possesses at least two distinct PTS phosphoryl transfer chains, EI(Ntr) --> NPr --> IIA(Ntr) and EI --> HPr --> IIA(sugar). Sequence comparisons allow prediction of residues likely to be important for specificity. This is the first report demonstrating specificity at the level of the energy coupling proteins of the PTS. PMID- 10473572 TI - Identification and molecular characterization of novel peroxidase with structural protein-like properties. AB - Elicitor treatment or mechanical damage to Scutellaria baicalensis Georgi (skullcap plants) callus causes an immediate insolubilization of a 36-kDa protein into cell walls. The 36-kDa protein was identified as peroxidase 1 by analysis of its internal amino acid sequence and by immunoblotting using affinity-purified anti-peroxidase 1. Insolubilized peroxidase 1 is cross-linked to lignin through covalent bonds, and the cross-linking is catalyzed in the presence of H(2)O(2) by peroxidase 1 itself. The properties of insolubilized peroxidase 1 resemble those of defense-related structural proteins (extensins and proline-rich proteins) cross-linked to cell wall. Although the isozymes peroxidases 2 and 3 have enzyme activities similar to peroxidase 1, they are not insolubilized by stress treatment. Molecular characterization established that peroxidase 1 contains regions characteristic of structural proteins, but peroxidases 2 and 3 do not have such regions. These results suggest that among the three isozymes, only peroxidase 1 has a structural protein-like function as well as an enzymatic function. PMID- 10473573 TI - Structural examination of autoregulation of multifunctional calcium/calmodulin dependent protein kinase II. AB - Regulation of Ca(2+)/calmodulin-dependent protein kinase II is likely based on an auto-inhibitory mechanism in which a segment of the kinase occupies the catalytic site in the absence of calmodulin. We analyze potential auto-inhibitory associations by employing charge reversal and hydrophobic-to-charged residue mutagenesis. We identify interacting amino acid pairs by using double mutants to test which modification in the catalytic domain complements a given change in the auto-inhibitory domain. Our studies identify the core pseudosubstrate sequence (residues 297-300) but reveal that distinct sequences centered about the autophosphorylation site at Thr-286 are involved in the critical auto-inhibitory interactions. Individual changes in any of the residues Arg-274, His-282, Arg 283, Lys-291, Arg-297, Phe-293, and Asn-294 in the auto-inhibitory domain or their interacting partners in the catalytic domain produces an enhanced affinity for calmodulin or generates a constitutively active enzyme. A structural model of Ca(2+)/calmodulin-dependent protein kinase II that incorporates these interactions shows that Thr-286 is oriented inwardly into a hydrophobic channel. The model explains why calmodulin must bind to the auto-inhibitory domain in order for Thr-286 in that domain to be phosphorylated and why introduction of phospho-Thr-286 produces the important Ca(2+)-independent state of the enzyme. PMID- 10473574 TI - Nerve growth factor (NGF)-induced calcium influx and intracellular calcium mobilization in 3T3 cells expressing NGF receptors. AB - The neurotrophins have been implicated in the acute regulation of synaptic plasticity. Neurotrophin-stimulated presynaptic calcium uptake appears to play a key role in this process. To understand the mechanism of neurotrophin-stimulated calcium uptake, the regulation of calcium uptake and intracellular mobilization by nerve growth factor (NGF) was investigated using NIH 3T3 cells stably transfected with either the high affinity NGF receptor p140(trk) (3T3-Trk) or the low affinity NGF receptor p75(NGFR) (3T3-p75). In 3T3-Trk cells, NGF increased both calcium uptake and intracellular calcium mobilization. In 3T3-p75 cells, NGF increased calcium uptake but not intracellular calcium mobilization. K-252a alone increased intracellular calcium in 3T3-Trk cells but not in 3T3-p75 cells. Nifedipine, an inhibitor of calcium uptake through L-type calcium channels, inhibited the action of NGF on both 3T3-Trk cells and 3T3-p75 cells, indicating that both p140(trk) and p75(NGFR) receptors are linked to nifedipine-sensitive L type calcium channels. These studies show that either NGF receptor will support increases in intracellular calcium but that p140(trk) does so by increasing both uptake and mobilization, whereas p75(NGFR) does so by increasing uptake only. PMID- 10473575 TI - Overexpression of catalase in cytosolic or mitochondrial compartment protects HepG2 cells against oxidative injury. AB - HepG2 cells were transfected with vectors containing human catalase cDNA and catalase cDNA with a mitochondrial leader sequence to allow comparison of the effectiveness of catalase overexpressed in the cytosolic or mitochondrial compartments to protect against oxidant-induced injury. Overexpression of catalase in cytosol and in mitochondria was confirmed by Western blot, and activity measurement and stable cell lines were established. The intracellular level of H(2)O(2) induced by exogenously added H(2)O(2) or antimycin A was lower in C33 cell lines overexpressing catalase in the cytosol and mC5 cell lines overexpressing catalase in the mitochondria as compared with Hp cell lines transfected with empty vector. Cell death caused by H(2)O(2), antimycin A, and menadione was considerably suppressed in both the mC5 and C33 cell lines. C33 and mC5 cells were also more resistant to apoptosis induced by H(2)O(2) and to the loss of mitochondrial membrane potential induced by H(2)O(2) and antimycin A. In view of the comparable protection by catalase overexpressed in the cytosol versus the mitochondria, catalase produced in both cellular compartments might act as a sink to decompose H(2)O(2) and move diffusable H(2)O(2) down its concentration gradient. The present study suggests that catalase in cytosol and catalase in mitochondria are capable of protecting HepG2 cells against cytotoxicity or apoptosis induced by oxidative stress. PMID- 10473576 TI - Dissecting the role of a conserved motif (the second region of homology) in the AAA family of ATPases. Site-directed mutagenesis of the ATP-dependent protease FtsH. AB - Escherichia coli FtsH is an ATP-dependent protease that belongs to the AAA protein family. The second region of homology (SRH) is a highly conserved motif among AAA family members and distinguishes these proteins in part from the wider family of Walker-type ATPases. Despite its conservation across the AAA family of proteins, very little is known concerning the function of the SRH. To address this question, we introduced point mutations systematically into the SRH of FtsH and studied the activities of the mutant proteins. Highly conserved amino acid residues within the SRH were found to be critical for the function of FtsH, with mutations at these positions leading to decreased or abolished ATPase activity. The effects of the mutations on the protease activity of FtsH correlated strikingly with their effects on the ATPase activity. The ATPase-deficient SRH mutants underwent an ATP-induced conformational change similar to wild type FtsH, suggesting an important role for the SRH in ATP hydrolysis but not ATP binding. Analysis of the data in the light of the crystal structure of the hexamerization domain of N-ethylmaleimide-sensitive fusion protein suggests a plausible mechanism of ATP hydrolysis by the AAA ATPases, which invokes an intermolecular catalytic role for the SRH. PMID- 10473577 TI - Identification of syntenin as a protein of the apical early endocytic compartment in Madin-Darby canine kidney cells. AB - We used flow cytometry to sort and analyze apical and basolateral endocytic vesicles from filter-grown Madin-Darby canine kidney (MDCK) cells after membrane internalization of the lipophilic fluorescent probe trimethylamino diphenylhexatriene. Western blot analysis of sorted fractions showed enrichment of the early endosomal markers transferrin receptor and the small GTPase Rab5. Two-dimensional gel analysis indicated that the apical and basolateral early endosomes differed significantly in their protein composition. We found nine polypeptides to be specifically enriched in apical or basolateral endocytic vesicles. An apical protein identified by microsequencing was the adaptor molecule syntenin. This protein contains two PDZ domains (PSD-95, Dlg, and ZO-1 homology) that bind syndecan and ephrin-B2 cytoplasmic domains. In MDCK cells, transiently overexpressed Myc-tagged syntenin localized to both plasma membrane domains and to an intracellular vesicular compartment. Syntenin positive vesicles colocalized with internalized transferrin in the perinuclear region. In addition, syntenin colocalized in the apical supranuclear region with Rab5 and Rab11; the latter is a marker for the apical recycling endosomes in MDCK cells. PMID- 10473578 TI - Shuttling of CTP:Phosphocholine cytidylyltransferase between the nucleus and endoplasmic reticulum accompanies the wave of phosphatidylcholine synthesis during the G(0) --> G(1) transition. AB - The transition from quiescence (G(0)) into the cell division cycle is marked by accelerated phospholipid turnover. We examined the rates of phosphatidylcholine (PC) synthesis and the activity, membrane affinity, and intracellular localization of the rate-limiting enzyme in the synthesis of PC, CTP:phosphocholine cytidylyltransferase (CT) during this transition. The addition of serum to quiescent IIC9 fibroblasts resulted in a wave of PC synthesis beginning at approximately 10 min, peaking at approximately 3 h with a >10-fold increase in rate, and declining to near basal rates by 10 h. CT activity, monitored in situ, was elevated approximately 3-fold between 1 and 2 h postserum. Neither CT mass nor its phosphorylation state changed during the surge in PC synthesis and CT activity. On the other hand, the ratio of particulate/soluble CT surged and then receded in concert with the wave of PC synthesis. During quiescence, CT was confined to the nucleus, as assessed by indirect immunofluorescence. Within 10 min after serum stimulation, a portion of the CT fluorescence appeared in the cytoplasm, where it intensified until approximately 4 h postserum. Thereafter, the cytoplasmic CT signal waned, while the nuclear signal increased, and by 8 h CT was once again predominantly nuclear. The dynamics of CT's apparent translocation in and out of the nucleus paralleled the wave of PC synthesis and the solubility changes of CT. Cytoplasmic CT co localized with BiP, a resident endoplasmic reticulum protein, in a double labeling experiment. These data suggest that the wave of PC synthesis that accompanies the G(0) --> G(1) transition is regulated by the coordinated changes in CT activity, membrane affinity, and intracellular distribution. We describe for the first time a redistribution of CT from the nucleus to the ER that correlates with an activation of the enzyme. We propose that this movement is required for the stimulation of PC synthesis during entry into the cell cycle. PMID- 10473580 TI - The nonclassical class I molecule CD1d associates with the novel CD8 ligand gp180 on intestinal epithelial cells. AB - Previous studies have shown that normal intestinal epithelial cells (IECs) are able to selectively activate CD8(+) T cells with suppressor activity, inducing proliferation associated with the activation of both the CD8-associated kinase p56(lck) and the T cell receptor (TCR)-associated kinase p59(fyn). This process appears to relate in part to a 180-kDa IEC surface glycoprotein, gp180, which binds to CD8 and activates CD8-associated p56(lck). However, purified gp180 alone is unable to induce T cell proliferation and does not activate p59(fyn). Because the class Ib molecule CD1d is expressed by IECs and monoclonal antibodies (mAbs) against CD1d inhibit IEC-induced proliferation of CD8(+) T cells, co immunoprecipitation and enzyme-linked immunosorbent assay studies were performed, which demonstrated an association of gp180 and CD1d on the IEC surface. Interestingly, the activation of p59(fyn) in IEC-T cell co-cultures was blocked by the anti-CD1d mAb D5 but not by the anti-gp180 mAb B9. Conversely, treatment of IECs with mAb B9 inhibited IEC-induced activation of p56(lck) but not p59(fyn). More directly, a human CD1d cDNA (FO-1 D5) transfectant was able to activate p59(fyn) but not p56(lck). These data suggest that the CD1d-gp180 complex on the surface of IECs can be recognized by the TCR-CD8 co-receptor, resulting in the activation of CD8(+) T cells. PMID- 10473579 TI - Cloning and expression of bovine neutrophil beta-defensins. Biosynthetic profile during neutrophilic maturation and localization of mature peptide to novel cytoplasmic dense granules. AB - beta-Defensins are microbicidal peptides implicated in host defense functions of phagocytic leukocytes and certain surface epithelial cells. Here we investigated the genetic structures and cellular expression of BNBD-4, -12, and -13, three prototypic bovine neutrophil beta-defensins. Characterization of the corresponding cDNAs indicated that BNBD-4 (41 residues) derives from a 63-amino acid prepropeptide and that BNBD-12 (38 residues) and BNBD-13 (42 residues) derive from a common 60-amino acid precursor (BNBD-12/13). The peptides were found to be encoded by two-exon genes that are closely related to bovine epithelial beta-defensin genes. BNBD-4 and BNBD-12/13 mRNAs were most abundant in bone marrow, but were expressed differentially in certain non-myeloid tissues. In situ hybridization and immunohistochemical studies demonstrated that BNBD-4 synthesis is completed early in myelopoiesis. BNBD-12 was localized exclusively to the novel dense granules, organelles that also contain precursors of cathelicidins, antimicrobial peptides that undergo proteolytic processing during phagocytosis. In contrast to cathelicidins, Western blot analyses revealed that mature beta-defensins are the predominant organellar form in myeloid cells. Stimulation of neutrophils with phorbol myristate acetate induced secretion of BNBD-12, indicating that it is co-secreted with pro-cathelicidins. The exocytosis of BNBD-12 by activated neutrophils reveals different mobilization pathways for myeloid alpha- and beta-defensins. PMID- 10473581 TI - A secreted form of the major histocompatibility complex class II-associated invariant chain inhibiting T cell activation. AB - Major histocompatibility complex (MHC) class II molecules function at the cell surface to present antigenic peptides to T helper cells. Intracellularly, MHC class II molecules are associated with the invariant chain (Ii). Ii can modulate MHC class II-dependent T cell activation through (i) assistance in the export of MHC class II molecules from the endoplasmic reticulum, (ii) providing a targeting signal for endosomal/lysosomal compartments, and (iii) preventing peptides from associating prematurely with MHC class II molecules. Here we describe the generation and subsequent secretion of a lumenal form of Ii, IiP25. IiP25 lacked the targeting sequences for transport to MHC class II compartments but contained part of the CLIP region that is known to compete with antigenic peptides for binding to MHC class II molecules. When added to an antigenic peptide presentation model system, IiP25 inhibited T cell activation by competing for the CLIP binding site at the plasma membrane. Secretion of a lumenal Ii fragment may represent an additional mechanism to modulate T cell activation by MHC class II molecules. PMID- 10473582 TI - Adherence of Borrelia burgdorferi. Identification of critical lysine residues in DbpA required for decorin binding. AB - Borrelia burgdorferi, the causative agent of Lyme disease, expresses on its surface two decorin binding adhesins, DbpA and DbpB. Previous studies have demonstrated that vaccination of mice with DbpA provided protection against challenge with heterologous Borrelia strains despite considerable sequence variability among DbpA in these strains. We have now examined the importance of individual amino acid residues in DbpA for decorin binding. We demonstrated that chemical modification of lysine residues resulted in loss of ligand binding activity. Of the 27 lysine residues in native DbpA from strain 297, 6 are present in most and 5 are conserved in all 30 DbpA sequences examined so far. Analysis of recombinant DbpA in which individual lysine residues have been mutated to alanine suggested that three of the conserved residues distributed throughout the DbpA sequence are required for decorin binding. These mutants lost their ability to bind decorin in Western ligand blot assay and bound reduced amounts of decorin in an ELISA. Furthermore, these mutant DbpA proteins did not inhibit the adherence of B. burgdorferi to a decorin substrata, and they did not recognize decorin in an extracellular matrix established by human fibroblast cultures. We conclude that the three lysine residues Lys-82, Lys-163, and Lys-170 are crucial for the binding of DbpA to decorin. PMID- 10473583 TI - Crystal structure of a maltogenic amylase provides insights into a catalytic versatility. AB - Amylases catalyze the hydrolysis of starch material and play central roles in carbohydrate metabolism. Compared with many different amylases that are able to hydrolyze only alpha-D-(1,4)-glycosidic bonds, maltogenic amylases exhibit catalytic versatility: hydrolysis of alpha-D-(1,4)- and alpha-D-(1,6)-glycosidic bonds and transglycosylation of oligosaccharides to C3-, C4-, or C6-hydroxyl groups of various acceptor mono- or disaccharides. It has been speculated that the catalytic property of the enzymes is linked to the additional approximately 130 residues at the N terminus that are absent in other typical alpha-amylases. The crystal structure of a maltogenic amylase from a Thermus strain was determined at 2.8 A. The structure, an analytical centrifugation, and a size exclusion column chromatography proved that the enzyme is a dimer in solution. The N-terminal segment of the enzyme folds into a distinct domain and comprises the enzyme active site together with the central (alpha/beta)(8) barrel of the adjacent subunit. The active site is a narrow and deep cleft suitable for binding cyclodextrins, which are the preferred substrates to other starch materials. At the bottom of the active site cleft, an extra space, absent in the other typical alpha-amylases, is present whose size is comparable with that of a disaccharide. The space is most likely to host an acceptor molecule for the transglycosylation and to allow binding of a branched oligosaccharide for hydrolysis of alpha-D (1,4)-glycosidic or alpha-D-(1,6)-glycosidic bond. The (alpha/beta)(8) barrel of the enzyme is the preserved scaffold in all the known amylases. The structure represents a novel example of how an enzyme acquires a different substrate profile and a catalytic versatility from a common active site and represents a framework for explaining the catalytic activities of transglycosylation and hydrolysis of alpha-D-(1,6)-glycosidic bond. PMID- 10473584 TI - Transmembrane tumor necrosis factor (TNF)-alpha inhibits adipocyte differentiation by selectively activating TNF receptor 1. AB - Tumor necrosis factor alpha (TNFalpha) is a potent cytokine with multiple biological activities and exists in two forms as follows: a 17-kDa soluble form that is a cleaved product of the 26-kDa transmembrane form (mTNFalpha). It has been suggested that the transmembrane form of TNFalpha is mainly responsible for localized responses via cell-cell contact. Here, we have examined the activities of transmembrane TNFalpha in cultured adipocytes. A non-cleavable transmembrane form of TNFalpha (mTNFDelta1-9K11E) was expressed in several preadipocyte cell lines using retroviral gene transfer. In wild type preadipocytes carrying both TNF receptors, expression of mTNFDelta1-9K11E resulted in inhibition of the differentiation program. The extent of this varied depending on the nature and strength of the adipogenic stimuli. The TNF receptor responsible for this function was determined by expressing mTNFDelta1-9K11E in preadipocyte cell lines lacking either TNF receptor 1 (TNFR1), 2 (TNFR2), or both. In order to confirm the results in the same cellular background, TNF receptors were also reconstituted in the cell lines lacking corresponding receptors. These experiments demonstrated that TNFR1 was necessary and sufficient for mediating mTNFDelta1-9K11E-induced inhibition of adipogenesis and that this action was similar to that of soluble TNFalpha. In conclusion, our results indicate that mTNFDelta1-9K11E is biologically active in cultured adipocytes and can alter the adipogenic program of these cells by selectively activating TNFR1. This may have physiological implications where local TNFalpha actions are thought to be generated at sites such as adipose tissue. PMID- 10473585 TI - Stereoselective carveol dehydrogenase from Rhodococcus erythropolis DCL14. A novel nicotinoprotein belonging to the short chain dehydrogenase/reductase superfamily. AB - A novel nicotinoprotein, catalyzing the dichlorophenolindophenol-dependent oxidation of carveol to carvone, was purified to homogeneity from Rhodococcus erythropolis DCL14. The enzyme is specifically induced after growth on limonene and carveol. Dichlorophenolindophenol-dependent carveol dehydrogenase (CDH) is a homotetramer of 120 kDa with each subunit containing a tightly bound NAD(H) molecule. The enzyme is optimally active at pH 5.5 and 50 degrees C and displays a broad substrate specificity with a preference for substituted cyclohexanols. When incubated with a diastereomeric mixture of (4R)- or (4S)-carveol, CDH stereoselectively catalyzes the conversion of the (6S)-carveol stereoisomers only. Kinetic studies with pure stereoisomers showed that this is due to large differences in V(max)/K(m) values and simultaneous product inhibition by (R)- or (S)-carvone. The R. erythropolis CDH gene (limC) was identified in an operon encoding the enzymes involved in limonene degradation. The CDH nucleotide sequence revealed an open reading frame of 831 base pairs encoding a 277-amino acid protein with a deduced mass of 29,531 Da. The CDH primary structure shares 10-30% sequence identity with members of the short chain dehydrogenase/reductase superfamily. Structure homology modeling with trihydroxynaphthalene reductase from Magnaporthe grisea suggests that CDH from R. erythropolis DCL14 is an alpha/beta one-domain protein with an extra loop insertion involved in NAD binding and a flexible C-terminal part involved in monoterpene binding. PMID- 10473586 TI - Protein kinase A is a negative regulator of renal branching morphogenesis and modulates inhibitory and stimulatory bone morphogenetic proteins. AB - Protein kinase A (PKA) regulates morphogenetic responses to bone morphogenetic proteins (BMPs) during embryogenesis. However, the mechanisms by which PKA regulates BMP function are unknown. During kidney development, BMP-2 and high doses of BMP-7 inhibit branching morphogenesis, whereas low doses of BMP-7 are stimulatory (Piscione, T. D., Yager, T. D., Gupta, I. R., Grinfeld, B., Pei, Y., Attisano, L., Wrana, J. L., and Rosenblum, N. D. (1997) Am. J. Physiol. 273, F961 F975). We examined the interactions between PKA and these BMPs in embryonic kidney explants and in the mouse inner medullary collecting duct-3 model of collecting duct morphogenesis. H-89, an inhibitor of PKA, stimulated branching morphogenesis and enhanced the stimulatory effect of low doses of BMP-7 on tubule formation. Furthermore, H-89 rescued the inhibition of tubulogenesis by BMP-2 (or high doses of BMP-7) by attenuating BMP-2-induced collecting duct apoptosis. In contrast, 8-bromo-cAMP, an activator of PKA, inhibited tubule formation and attenuated the stimulatory effects of low doses of BMP-7. To determine mechanisms underlying the interdependence of BMP signaling and PKA activity, we examined the effect of PKA on the known signaling events in the BMP-2-dependent Smad1 signaling pathway and the effect of BMP-2 on PKA activity. PKA did not induce endogenous Smad1 phosphorylation, Smad1-Smad4 complex formation, or Smad1 nuclear translocation. In contrast, BMP-2 increased endogenous PKA activity and induced phosphorylation of the PKA effector, cAMP-response element-binding protein, in a PKA-dependent manner. We conclude that BMP-2 induces activation of PKA and that PKA regulates the effects of BMPs on collecting duct morphogenesis without activating the known signaling events in the BMP-2-dependent Smad1 signaling pathway. PMID- 10473587 TI - Identification and purification of a novel receptor for secretory phospholipase A(2) in porcine cerebral cortex. AB - A specific phospholipase A(2) receptor from porcine cerebral cortex has been characterized (K(d) = 145 nM, B(max) = 0.4 pmol/mg membrane protein) by using a radioiodinated derivative of ammodytoxin C (AtxC), a snake venom presynaptically neurotoxic group IIA phospholipase A(2). After the receptor was solubilized in a ligand-binding form, it was approximately 14,000-fold enriched by chromatography on wheat germ lectin-Sepharose and AtxC-Affi-Gel 10. The receptor is a single chain glycoprotein with an apparent molecular mass of 180 kDa and binds toxic and non-toxic phospholipases A(2) of either group I or II. It also recognizes conjugates of bovine serum albumin with mannose, N-acetylglucosamine, and galactose. In its molecular mass and pharmacological profile, the AtxC receptor resembles the M-type receptor for secretory phospholipases A(2) from rabbit skeletal muscle (a C-type multilectin, homologous to macrophage mannose receptor), yet in terms of relative abundance in brain and antigenicity, these two receptors are completely different. A further AtxC receptor of approximately 200 kDa discovered in porcine liver was, however, recognized by anti-rabbit M type phospholipase A(2) receptor antibodies. There are, therefore, two immunologically distinct secretory phospholipase A(2) receptors of about 200 kDa in the same species. Although the liver receptor is related to the M-type secretory phospholipase A(2) receptors, the brain receptor is not and belongs to a novel group of secretory phospholipase A(2) receptors. PMID- 10473588 TI - Binding of p53 to the KIX domain of CREB binding protein. A potential link to human T-cell leukemia virus, type I-associated leukemogenesis. AB - The pleiotropic cellular coactivator CREB binding protein (CBP) plays a critical role in supporting p53-dependent tumor suppressor functions. p53 has been shown to directly interact with a carboxyl-terminal region of CBP for recruitment of the coactivator to p53-responsive genes. In this report, we identify the KIX domain as a new p53 contact point on CBP. We show that both recombinant and endogenous forms of p53 specifically interact with KIX. We demonstrate that the activation domain of p53 participates in KIX binding and provide evidence showing that this interaction is critical for p53 transactivation function. The human T cell leukemia virus, type-I-encoded oncoprotein Tax is a well established repressor of p53 transcription function. Like p53, Tax also binds to KIX. The finding that both transcription factors bind to a common region of CBP suggests that coactivator competition may account for the observed repression. We demonstrate reciprocal repression between Tax and p53 in transient transfection assays, supporting the idea of intracellular coactivator competition. We biochemically confirm coactivator competition by directly showing that both transcription factors bind to KIX in a mutually exclusive fashion. These data provide molecular evidence for the observed intracellular competition and suggest that Tax inhibits p53 function by abrogating a novel p53-KIX interaction. Thus, Tax competition for the p53-KIX complex may be a pivotal event in the human T cell leukemia virus, type I transformation pathway. PMID- 10473589 TI - Soluble Rous sarcoma virus reverse transcriptases alpha, alphabeta, and beta purified from insect cells are processive DNA polymerases that lack an RNase H 3' --> 5' directed processing activity. AB - Reverse transcriptase (RT) isolated from Rous sarcoma virus (RSV) consists of heterodimeric RTalphabeta, RTalpha, and RTbeta. The alpha subunit (63 kDa) contains an N-terminal polymerase and a C-terminal RNase H domain. The N terminus of beta (95 kDa) corresponds to alpha with the integrase domain attached to the C terminus (32 kDa). We have constructed baculoviruses expressing the genes for alpha or beta or the entire pol (99 kDa). Infection of insect cells with recombinant virus yielded highly active and soluble RSV RT enzymes that could be purified to >90% homogeneity. HPLC gel filtration showed that alpha is a dimeric enzyme that can be partially monomerized upon the addition of 45% Me(2)SO. DNA synthesis on DNA-DNA and DNA-RNA primer-templates in the presence of competitor substrates revealed that alphabeta and beta as well as alpha are processive polymerases. However, the affinity of beta and alphabeta for primer-template substrates appears to be higher than that of alpha. All RSV enzymes investigated have the potential to displace RNA-RNA duplexes more efficiently than human immunodeficiency virus type 1 RT. Unlike human immunodeficiency virus type 1 RT, RSV RTs can catalyze an initial RNase H endonucleolytic cleavage of the RNA template but not a 3' --> 5' directed processing activity. PMID- 10473590 TI - Nitric oxide modulates beta(2)-adrenergic receptor palmitoylation and signaling. AB - To determine whether nitric oxide (NO) modulates the beta-adrenergic signaling pathway, we treated cells expressing beta(2)-adrenergic receptors (beta(2)AR) with the NO donors, 3-morpholinosydnonimine (SIN-1) and 1,2,3,4-oxatriazolium, 5 amino-3-(3-chloro-2-methylphenyl)chloride and determined the intracellular production of cAMP after exposure to beta-adrenergic receptor agonists, cholera toxin and forskolin. NO significantly decreased the potency of the beta adrenergic agonist, isoproterenol, to stimulate cAMP production without affecting the stimulatory action of forskolin and cholera toxin, which directly activate adenylyl cyclase and G(s), respectively. Treatment with the NO donor increased the guanyl nucleotide-sensitive high affinity constant for the agonist, isoproterenol, thus suggesting that it reduced functional coupling between the receptor and G(s). Stimulation of endogenous NO production by lipopolysaccharide in RAW 264.7 macrophages also caused a significant increase in the EC(50) for isoproterenol-stimulated cAMP production. SIN-1 treatment also led to a reduction in both basal and isoproterenol-stimulated incorporation of [(3)H]palmitate into the beta(2)AR. Signaling through the nonpalmitoylated, Gly(341)beta(2)AR mutant was unchanged by SIN-1 treatment. Given the link between beta(2)AR palmitoylation and its responsiveness to agonist, these results suggest that the primary action of NO was depalmitoylation of the beta(2)AR resulting in decreased signaling through the beta(2)AR. PMID- 10473591 TI - Structural and functional characterization of H protein mutants of the glycine decarboxylase complex. AB - The mitochondrial glycine decarboxylase complex (GDC) consists of four component enzymes (P, H, T, and L proteins) involved in the breakdown of glycine. In order to investigate structural interactions involved in the stabilization of the methylamine-loaded H protein (a transient species in the GDC reaction), we designed several mutants of H apoprotein. Structural analysis of the wild-type and mutants of H apoprotein emphasized the necessity to carefully assess, by biophysical techniques, the correct folding of mutated proteins prior to investigate their biochemical properties. The correctly folded wild-type and mutants of H apoprotein were in vitro lipoylated and then characterized in the context of GDC reaction by studying the reconstituted complex and partial reactions. We showed that Val(62) and Ala(64), surrounding the lipoyl-lysine, play an important role in the molecular events that govern the reaction between P and H protein but do not intervene in the recognition of the binding site of lipoic acid by lipoyl ligase. The biochemical results obtained with the HE14A mutant of H protein pointed out the major role of the Glu(14) amino acid residue in the GDC catalysis and highlighted the importance of the ionic and hydrogen bounds in the hydrophobic cleft of H protein for the stabilization of the methylamine-loaded lipoyl arm. PMID- 10473592 TI - Regulation of cAMP-mediated signal transduction via interaction of caveolins with the catalytic subunit of protein kinase A. AB - cAMP-dependent processes are essential for cell growth, differentiation, and homeostasis. The classic components of this system include the serpentine receptors, heterotrimeric G-proteins, adenylyl cyclase, protein kinase A (PKA), and numerous downstream target substrates. Evidence is accumulating that some members of this cascade are concentrated within membrane microdomains, termed caveolae and caveolae-related domains. In addition, the caveolin-1 protein has been shown to interact with some of these components, and this interaction inhibits their enzymatic activity. However, the functional effects of caveolins on cAMP-mediated signaling at the most pivotal step, PKA activation, remain unknown. Here, we show that caveolin-1 can dramatically inhibit cAMP-dependent signaling in vivo. We provide evidence for a direct interaction between caveolin 1 and the catalytic subunit of PKA both in vitro and in vivo. Caveolin-1 binding appears to be mediated both by the caveolin scaffolding domain (residues 82-101) and a portion of the C-terminal domain (residues 135-156). Further functional analysis indicates that caveolin-based peptides derived from these binding regions can inhibit the catalytic activity of purified PKA in vitro. Mutational analysis of the caveolin scaffolding domain reveals that a series of aromatic residues within the caveolin scaffolding domain are critical for mediating inhibition of PKA. In addition, co-expression of caveolin-1 and PKA in cultured cells results in their co-localization as seen by immunofluorescence microscopy. In cells co-expressing caveolin-1 and PKA, PKA assumed a punctate distribution that coincided with the distribution of caveolin-1. In contrast, in cells expressing PKA alone, PKA was localized throughout the cytoplasm and yielded a diffuse staining pattern. Taken together, our results suggest that the direct inhibition of PKA by caveolin-1 is an important and previously unrecognized mechanism for modulating cAMP-mediated signaling. PMID- 10473593 TI - Chicken macrophage stimulating protein is a ligand of the receptor protein tyrosine kinase Sea. AB - Affinity chromatography, employing the extracellular domain of the Sea receptor, was used to enrich Sea-binding proteins from chicken serum. One isolated protein bound both a Sea-immunoglobulin fusion protein and an antisera raised against murine macrophage stimulating protein. Amino-terminal sequencing of the dual reactive protein yielded sequences which were identical to the predicted alpha and beta subunits of chicken macrophage stimulating protein. The partially purified chicken macrophage stimulating protein caused autophosphorylation of the Sea receptor. Previous work showed that recombinant expression of fully activatible human or mouse macrophage stimulating protein required a specific Cys to Ala substitution (Wahl, R. C., Costigan, V. J., Batac, J. P., Chen, K., Cam, L., Courchesne, P. L., Patterson, S. D. Zhang, K., and Pacifici, R. E. (1997) J. Biol. Chem. 272, 1-4). Therefore, we expressed both the wild type and the specific Cys to Ala form of chicken macrophage stimulating protein as recombinant proteins. After proteolytic activation, only conditioned media from COS cells transfected with the C665A chicken macrophage stimulating protein, but not from wild type chicken macrophage-stimulating protein, or control vector, was detected by the Sea-immunoglobulin fusion protein in Western blotting experiments. Conditioned media containing the C665A chicken macrophage-stimulating protein readily caused Sea phosphorylation, while conditioned media containing the wild type chicken macrophage-stimulating protein was only effective at inducing receptor phosphorylation at high concentrations. In addition to receptor phosphorylation, the C665A chicken macrophage-stimulating protein induced phosphorylation of Shc, Erk1, and Erk 2. We conclude that macrophage-stimulating protein is a ligand of the Sea receptor protein-tyrosine kinase. PMID- 10473594 TI - Antiproliferative activity of G-rich oligonucleotides correlates with protein binding. AB - Oligonucleotides have been extensively studied as antisense or antigene agents that can potentially modulate the expression of specific genes. These strategies rely on sequence-specific hybridization of the oligonucleotide to mRNA or genomic DNA. Recently, it has become clear that oligonucleotides often have biological activities that cannot be attributed to their sequence-specific interactions with nucleic acids. Here we describe a series of guanosine-rich phosphodiester oligodeoxynucleotides that strongly inhibit proliferation in a number of human tumor cell lines. The presence of G-quartets in the active oligonucleotides is demonstrated using an UV melting technique. We show that G-rich oligonucleotides bind to a specific cellular protein and that the biological activity of the oligonucleotides correlates with binding to this protein. The G-rich oligonucleotide-binding protein was detected in both nuclear and cytoplasmic extracts and in proteins derived from the plasma membrane of cells. We present strong evidence that this protein is nucleolin, a multifunctional phosphoprotein whose levels are related to the rate of cell proliferation. Our results indicate that binding of G-rich oligonucleotides to nucleolin may be responsible for their non-sequence-specific effects. Furthermore, these oligonucleotides represent a new class of potentially therapeutic agents with a novel mechanism of action. PMID- 10473595 TI - Inactivation of protein-tyrosine phosphatases as mechanism of UV-induced signal transduction. AB - UV irradiation of cells causes ligand-independent activation of receptor tyrosine kinases. On the basis of dephosphorylation kinetics, UV-induced inactivation of receptor-directed tyrosine phosphatases (PTP) has been proposed as the mechanism of receptor activation (Knebel, A., Rahmsdorf, H. J., Ullrich, A., and Herrlich, P. (1996) EMBO J. 15, 5314-5325). Here we show that four defined protein-tyrosine phosphatases (PTPs), SHP-1, RPTPalpha, RPTPsigma, and DEP-1, are partially inactivated upon UV irradiation of PTP-overexpressing cells. The dephosphorylation of coexpressed platelet-derived growth factor beta (PDGFbeta) receptor by RPTPalpha is inhibited upon UV irradiation. UV converts RPTPalpha into a substrate-trapping enzyme which can coprecipitate PDGFbeta receptor, similarly to the PTP mutant at the active-center cysteine: C433S. In agreement with the proposed mechanism that inactivation of PTPs accounts for receptor tyrosine kinase activation, no evidence for a UV-induced receptor cross-linking could be obtained in PDGFbeta receptor-enriched membrane micelle preparations and in PDGFbeta receptor overexpressing 293 cells. The intrinsic activity of PDGFbeta receptor kinase was required for the UV-induced enhancement of receptor phosphorylation, but was not changed upon UV irradiation. The data support a mechanism of UV-induced signal transduction involving inactivation of PTPs through an unknown reactive intermediate that oxidizes the conserved cysteine in the active sites of PTPs. PMID- 10473596 TI - Cig30 and Pitx3 genes are arranged in a partially overlapping tail-to-tail array resulting in complementary transcripts. AB - The mouse Cig30 gene codes for a 30-kDa membrane glycoprotein, which appears to have a role in the recruitment of brown adipose tissue. To elucidate the structure of the Cig30 gene, we have isolated a lambda phage genomic DNA clone containing the entire mouse gene and found that Cig30 consists of four exons that are spread over 4 kilobase pairs of genomic sequence. Using a fluorescence in situ hybridization assay and interspecific backcross panel mapping, we have localized the Cig30 locus to the distal region of mouse chromosome 19, between the Tlx1 and Ins1 loci. Sequencing of the corresponding lambda clone to completion revealed that the insert contained yet another gene in the opposite orientation. It turned out to be the newly identified homeobox gene Pitx3. Interestingly, the genes are very tightly linked, so that the 3' ends of their transcripts are complementary. Thus, our results provide evidence for bidirectional transcription of a several hundred base pair-long DNA region as a result of the extremely tight linkage between Cig30 and Pitx3. PMID- 10473597 TI - Macrophage colony-stimulating factor promotes cell survival through Akt/protein kinase B. AB - The signaling pathways activated by the macrophage colony-stimulating factor (M CSF) to promote survival of monocyte and macrophage lineage cells are not well established. In an effort to elucidate these pathways, we have used two cell types responsive to M-CSF: NIH 3T3 fibroblasts genetically engineered to express human M-CSF receptors (3T3-FMS cells) and human monocytes. M-CSF treatment induced M-CSF receptor tyrosine phosphorylation and recruitment of the p85 subunit of phosphatidylinositol 3-kinase (PI3K) to these receptors. These M-CSF receptor events correlated with activation of the serine/threonine kinase Akt. To clarify that PI3K products activate Akt in response to M-CSF, NIH 3T3 fibroblasts expressing mutant human M-CSF receptors (3T3-FMS(Y809F)) that fail to activate Ras in response to M-CSF also exhibit increased Akt kinase activity in response to M-CSF challenge. Furthermore, Akt appears to be the primary regulator of survival in 3T3-FMS cells, as transfection of genes encoding dominant-negative Akt isoforms into these fibroblasts blocked M-CSF-induced survival. In normal human monocytes, M-CSF increased the levels of tyrosine-phosphorylated proteins and induced Akt activation in a PI3K-dependent manner. The PI3K inhibitor LY294002 blocked M-CSF-mediated monocyte survival, an effect that was partially restored by caspase-9 inhibitors. These data suggest that M-CSF may induce cell survival through Akt-induced suppression of caspase-9 activation. PMID- 10473598 TI - Characterization of the regulatory domains of the human skn-1a/Epoc-1/Oct-11 POU transcription factor. AB - The Skn-1a POU transcription factor is primarily expressed in keratinocytes of murine embryonic and adult epidermis. Although some POU factors expressed in a tissue-specific manner are important for normal differentiation, the biological function of Skn-1a remains unknown. Previous in vitro studies indicate that Skn 1a has the ability to transactivate markers of keratinocyte differentiation. In this study, we have characterized Skn-1a's transactivation domain(s) and engineered a dominant negative protein that lacked this transactivation domain. Deletional analysis of the human homologue of Skn-1a with three target promoters revealed the presence of two functional domains: a primary C-terminal transactivation domain and a combined N-terminal inhibitory domain and transactivation domain. Skn-1a lacking the C-terminal region completely lost transactivation ability, irrespective of the promoter tested, and was able to block transactivation by normal Skn-1a in competition assays. Compared with full length, Skn-1a lacking the N-terminal region demonstrated either increased transactivation (bovine cytokeratin 6 promoter), comparable transactivation (human papillomavirus type 1a long control region), or loss of transactivation (human papillomavirus type 18 long control region). The identification of a primary C-terminal transactivation domain enabled us to generate a dominant negative Skn-1a factor, which will be useful in the quest for a better understanding of this keratinocyte-specific gene regulator. PMID- 10473599 TI - The release of acetylcholine receptor inducing activity (ARIA) from its transmembrane precursor in transfected fibroblasts. AB - Acetylcholine receptor inducing activity (ARIA) is made by motoneurons and is released at the neuromuscular synapse to stimulate the synthesis of acetylcholine receptors by skeletal muscle. ARIA is derived from a transmembrane precursor (pro ARIA) via proteolytic cleavage of the ectodomain. We studied requirements in the amino acid sequence at the cleavage site with various substitution and deletion mutations. Wild type (WT) and mutant proteins were transiently expressed in COS cells, and release of ARIA into the conditioned medium was measured by tyrosine phosphorylation of its receptor, p185, in L6 cells. Removal of all potential cleavage sites between the extracellular epidermal growth factor domain and the transmembrane domain by substitution and small deletions (<11 amino acid residues out of 21) did not significantly reduce ARIA release, whereas larger deletions abolished it. We propose that cleavage occurs independently of amino acid sequence at a short distance from the epidermal growth factor domain, unless sterically hindered by the nearby secondary structure. A mutant with shorter cytoplasmic domain ("c" isoform) released significantly less ARIA than the WT ("a" isoform), suggesting that the c isoform may be suitable for signaling through direct cell-cell contact. Alternatively, proteolytic conversion of the a isoform to the c isoform may rapidly down-regulate release of ARIA. PMID- 10473600 TI - Canalicular export pumps traffic with polymeric immunoglobulin A receptor on the same microtubule-associated vesicle in rat liver. AB - Basolateral to apical vesicular transcytosis in the hepatocyte is an essential pathway for the delivery of compounds from the sinusoidal blood to the bile and to traffic newly synthesized resident apical membrane proteins to their site of function at the canalicular membrane front. To characterize this pathway better, microtubules in a hepatocyte homogenate were polymerized by addition of taxol, and associated membrane-bound vesicles were isolated. This fraction was enriched in polymeric immunoglobulin A receptor and contained apical membrane proteins. Immunoelectron microscopy demonstrated that polymeric immunoglobulin A receptor was localized predominantly on vesicles ranging from 100 to 160 nm and that the multidrug resistance protein 2 and the bile salt export pump co-localized on these vesicles. The minus-ended microtubule motor, dynein, was highly enriched in the fraction, and its intermediate chain could be released effectively by incubation with 1 mM ATP or GTP. However, the association of the transcytotic vesicles with the microtubules was not sensitive to hydrolyzable or non hydrolyzable nucleotides. This study characterizes a fraction of microtubule associated vesicles from rat hepatocytes and demonstrates that several resident apical membrane transport proteins and the polymeric immunoglobulin A receptor traffic on the same vesicle. PMID- 10473601 TI - Antizyme2 is a negative regulator of ornithine decarboxylase and polyamine transport. AB - The antizyme family consists of closely homologous proteins believed to regulate cellular polyamine pools. Antizyme1, the first described, negatively regulates ornithine decarboxylase, the initial enzyme in the biosynthetic pathway for polyamines. Antizyme1 targets ornithine decarboxylase for degradation and inhibits polyamine transport into cells, thereby diminishing polyamine pools. A polyamine-stimulated ribosomal frameshift is required for decoding antizyme1 mRNA. Recently, additional novel conserved members of the antizyme family have been described. We report here the properties of one of these, antizyme2. Antizyme2, like antizyme1, binds to ornithine decarboxylase and inhibits polyamine transport. Using a baculovirus expression system in cultured Sf21 insect cells, both antizymes were found to accelerate ornithine decarboxylase degradation. Expression of either antizyme1 or 2 in Sf21 cells also diminished their uptake of the polyamine spermidine. Both forms of antizyme can therefore function as negative regulators of polyamine production and transport. However, in contrast to antizyme1, antizyme2 has negligible ability to stimulate degradation of ornithine decarboxylase in a rabbit reticulocyte lysate. PMID- 10473602 TI - Progesterone stimulation of human insulin-like growth factor-binding protein-5 gene transcription in human osteoblasts is mediated by a CACCC sequence in the proximal promoter. AB - Insulin-like growth factor-binding protein-5 (IGFBP-5) is produced by osteoblasts and potentiates insulin-like growth factor mitogenic stimulation in osteoblast cell cultures. Progesterone (PG) increased IGFBP-5 expression in normal human osteoblasts and increased IGFBP-5 transcription in U2 human osteosarcoma cells. We developed a chloramphenicol acetyltransferase reporter construct containing the human IGFBP-5 proximal promoter sequence, which includes TATA and CAAT boxes, and five putative PG response element half-sites. 10(-8) M PG increased promoter activity of this construct in U2 cells co-transfected with a PG receptor isoform A (PR(A)) expression vector. Analysis of 5' deletion constructs indicates that PG transactivation of IGFBP-5 promoter activity does not require the PG response element half-sites but does require the region -162 to -124 containing two tandem CACCC box sequences. Mutation of the proximal CACCC box at -139 eliminated PG transactivation. Gel shift assays using a -162 to -124 DNA fragment, U2 cell nuclear extracts, and purified PR(A) protein indicate that nuclear factors bind to a CACCC sequence at -139 and that PR(A) alters the pattern of transcription factor interaction with the CACCC sequence. Using a luciferase reporter construct containing base pairs -252 to +24 of the IGFBP-5 promoter, we found that both PR(A) and PR(B) isoforms mediated PG stimulation of promoter activity. These results suggest that PG may stimulate IGFBP-5 gene transcription via a novel mechanism involving PR and CACCC-binding factors. PMID- 10473603 TI - Clinical severity and thermodynamic effects of iron-responsive element mutations in hereditary hyperferritinemia-cataract syndrome. AB - Hereditary hyperferritinemia-cataract syndrome (HHCS) is a novel genetic disorder characterized by elevated serum ferritin and early onset cataract formation. The excessive ferritin production in HHCS patients arises from aberrant regulation of L-ferritin translation caused by mutations within the iron-responsive element (IRE) of the L-ferritin transcript. IREs serve as binding sites for iron regulatory proteins (IRPs), iron-sensing proteins that regulate ferritin translation. Previous observations suggested that each unique HHCS mutation conferred a characteristic degree of hyperferritinemia and cataract severity in affected individuals. Here we have measured the in vitro affinity of the IRPs for the mutant IREs and correlated decreases in binding affinity with clinical severity. Thermodynamic analysis of these IREs has also revealed that although some HHCS mutations lead to changes in the stability and secondary structure of the IRE, others appear to disrupt IRP-IRE recognition with minimal effect on IRE stability. HHCS is a noteworthy example of a human genetic disorder that arises from mutations within a protein-binding site of an mRNA cis-acting element. Analysis of the effects of these mutations on the energetics of the RNA-protein interaction explains the phenotypic variabilities of the disease state. PMID- 10473604 TI - Inhibition of interleukin-1-stimulated NF-kappaB RelA/p65 phosphorylation by mesalamine is accompanied by decreased transcriptional activity. AB - Nuclear factor kappaB (NF-kappaB) is an inducible transcription factor that regulates genes important in immunity and inflammation. The activity of NF-kappaB is highly regulated: transcriptionally active NF-kappaB proteins are sequestered in the cytoplasm by inhibitory proteins, IkappaB. A variety of extracellular signals, including interleukin-1 (IL-1), activate NF-kappaB by inducing phosphorylation and degradation of IkappaB, allowing nuclear translocation and DNA binding of NF-kappaB. Many of the stimuli that activate NF-kappaB by inducing IkappaB degradation also cause phosphorylation of the NF-kappaB RelA (p65) polypeptide. The transactivating capacity of RelA is positively regulated by phosphorylation, suggesting that in addition to cytosolic sequestration by IkappaB, phosphorylation represents another mechanism for control of NF-kappaB activity. In this report, we demonstrate that mesalamine, an anti-inflammatory aminosalicylate, dose-dependently inhibits IL-1-stimulated NF-kappaB-dependent transcription without preventing IkappaB degradation or nuclear translocation and DNA binding of the transcriptionally active NF-kappaB proteins, RelA, c-Rel, or RelB. Mesalamine was found to inhibit IL-1-stimulated RelA phosphorylation. These data suggest that pharmacologic modulation of the phosphorylation status of RelA regulates the transcriptional activity of NF-kappaB, independent of nuclear translocation and DNA binding. These findings highlight the importance of inducible phosphorylation of RelA in the control of NF-kappaB activity. PMID- 10473605 TI - Nucleotide-regulated calcium signaling in lung fibroblasts and epithelial cells from normal and P2Y(2) receptor (-/-) mice. AB - To test for the role of the P2Y(2) receptor (P2Y(2)-R) in the regulation of nucleotide-promoted Ca(2+) signaling in the lung, we generated P2Y(2)-R-deficient (P2Y(2)-R(-/-)) mice and measured intracellular Ca(2+)(i) responses (DeltaCa(2+)(i)) to nucleotides in cultured lung fibroblasts and nasal and tracheal epithelial cells from wild type and P2Y(2)-R(-/-) mice. In the wild type fibroblasts, the rank order of potencies for nucleotide-induced DeltaCa(2+)(i) was as follows: UTP >/= ATP >> ADP > UDP. The responses induced by these agonists were completely absent in the P2Y(2)-R(-/-) fibroblasts. Inositol phosphate responses paralleled those of DeltaCa(2+)(i) in both groups. ATP and UTP also induced Ca(2+)(i) responses in wild type airway epithelial cells. In the P2Y(2) R(-/-) airway epithelial cells, UTP was ineffective. A small fraction (25%) of the ATP response persisted. Adenosine and alpha,beta-methylene ATP were ineffective, and ATP responses were not affected by adenosine deaminase or by removal of extracellular Ca(2+), indicating that neither P1 nor P2X receptors mediated this residual ATP response. In contrast, 2-methylthio-ADP promoted a substantial Ca(2+)(i) response in P2Y(2)-R(-/-) cells, which was inhibited by the P2Y(1) receptor antagonist adenosine 3'-5'-diphosphate. These studies demonstrate that P2Y(2)-R is the dominant purinoceptor in airway epithelial cells, which also express a P2Y(1) receptor, and that the P2Y(2)-R is the sole purinergic receptor subtype mediating nucleotide-induced inositol lipid hydrolysis and Ca(2+) mobilization in mouse lung fibroblasts. PMID- 10473606 TI - Effect of loss of P2Y(2) receptor gene expression on nucleotide regulation of murine epithelial Cl(-) transport. AB - Extracellular nucleotides are believed to be important regulators of ion transport in epithelial tissues as a result of their ability to activate cell surface receptors. Although numerous receptors that bind nucleotides have been identified, the complexity of this receptor family, combined with the lack of pharmacological agents specific for these receptors, has made the assignment of particular receptors and ligands to physiological responses difficult. Because ATP and UTP appear equipotent and equieffective in regulating ion transport in many epithelia, we tested the hypothesis that the P2Y(2) receptor (P2Y(2)-R) subtype mediates these responses in mouse epithelia, with gene targeting techniques. Mice with the P2Y(2)-R locus targeted and inactivated (P2Y(2)-R(-/-)) were generated, airways (trachea), gallbladder, and intestines (jejunum) excised, and Cl(-) secretory responses to luminal nucleotide additions measured in Ussing chambers. Comparison of P2Y(2)-R(+/+) with P2Y(2)-R(-/-) mice revealed that P2Y(2)-R mediated most (>85-95%) nucleotide-stimulated Cl(-) secretion in trachea, about 50% of nucleotide responses in the gallbladder, and none of the responses in the jejunum. Dose-effect relationships for nucleotides in tissues from P2Y(2)-R(-/-) mice suggest that the P2Y(6)-R regulates ion transport in gallbladder and to a lesser extent trachea, whereas P2Y(4) and/or unidentified receptor(s) regulate ion transport in jejunum. We conclude that the P2Y(2) receptor is the dominant P2Y purinoceptor that regulates airway epithelial ion transport, whereas other P2Y receptor subtypes are relatively more important in other nonrespiratory epithelia. PMID- 10473608 TI - Functional characterization of the promoter of the X-linked ectodermal dysplasia gene. AB - Anhidrotic ectodermal dysplasia (EDA) is a disorder characterized by poor development of hair, teeth, and sweat glands, and results from lesions in the X linked EDA gene. We have cloned a 1.6-kilobase 5'-flanking region of the human EDA gene and used it to analyze features of transcriptional regulation. Primer extension analysis located a single transcription initiation site 264 base pairs (bp) upstream of the translation start site. When the intact cloned fragment or truncated derivatives were placed upstream of a reporter luciferase gene and transfected into a series of cultured cells, expression comparable with that conferred by an SV40 promoter-enhancer was observed. The region lacks a TATA box sequence, and basal transcription from the unique start site is dependent on two binding sites for the Sp1 transcription factor. One site lies 38 bp 5' to the transcription start site, in a 71-bp sequence that is sufficient to support up to 35% of maximal transcription. The functional importance of the Sp1 sites was demonstrated when cotransfection of an Sp1 expression vector transactivated the EDA promoter in the SL2 Drosophila cell line that otherwise lacks endogenous Sp1. Also, both Sp1 binding sites were active in footprinting and gel shift assays in the presence of either crude HeLa cell nuclear extract or purified Sp1 and lost activity when the binding sites were mutated. A second region involved in positive control was localized to a 40-bp sequence between -673 and -633 bp. This region activated an SV40 minimal promoter 4- to 5-fold in an orientation independent manner and is thus inferred to contain an enhancer region. PMID- 10473607 TI - Calmodulin and protein kinase C increase Ca(2+)-stimulated secretion by modulating membrane-attached exocytic machinery. AB - The molecular mechanisms underlying the Ca(2+) regulation of hormone and neurotransmitter release are largely unknown. Using a reconstituted [(3)H]norepinephrine release assay in permeabilized PC12 cells, we found that essential proteins that support the triggering stage of Ca(2+)-stimulated exocytosis are enriched in an EGTA extract of brain membranes. Fractionation of this extract allowed purification of two factors that stimulate secretion in the absence of any other cytosolic proteins. These are calmodulin and protein kinase Calpha (PKCalpha). Their effects on secretion were confirmed using commercial and recombinant proteins. Calmodulin enhances secretion in the absence of ATP, whereas PKC requires ATP to increase secretion, suggesting that phosphorylation is involved in PKC- but not calmodulin-mediated stimulation. Both proteins modulate release events that occur in the triggering stage of exocytosis. The half-maximal increase was elicited by 3 nM PKC and 75 nM calmodulin. These results suggest that calmodulin and PKC increase Ca(2+)-activated exocytosis by directly modulating the membrane- or cytoskeleton-attached exocytic machinery downstream of Ca(2+) elevation. PMID- 10473609 TI - SH2-B, a membrane-associated adapter, is phosphorylated on multiple serines/threonines in response to nerve growth factor by kinases within the MEK/ERK cascade. AB - SH2-B has been shown to be required for nerve growth factor (NGF)-mediated neuronal differentiation and survival, associate with NGF receptor TrkA, and be tyrosyl-phosphorylated in response to NGF. In this work, we examined whether NGF stimulates phosphorylation of SH2-B on serines/threonines. NGF promotes a dramatic upward shift in mobility of SH2-B, resulting in multiple forms that cannot be attributed to tyrosyl phosphorylation. Treatment of SH2-B with protein phosphatase 2A, a serine/threonine phosphatase, reduces the many forms to two. PD98059, a MEK inhibitor, dramatically inhibits NGF-promoted phosphorylation of SH2-B on serines/threonines, whereas depletion of 4beta-phorbol 12-myristate 13 acetate-sensitive protein kinase Cs does not. ERKs 1 and 2 phosphorylate SH2 Bbeta primarily on Ser-96 in vitro. However, NGF still stimulates serine/threonine phosphorylation of SH2-Bbeta(S96A). SH2-Bbeta(S96A), like wild type SH2-Bbeta, enhances NGF-induced neurite outgrowth. In contrast, SH2 Bbeta(R555E) containing a defective SH2 domain blocks NGF-induced neurite outgrowth and displays greatly reduced phosphorylation on serines/threonines in response to NGF. SH2-Bbeta(R555E), like wild-type SH2-Bbeta, associates with the plasma membrane, suggesting that the dominant negative effect of SH2-Bbeta(R555E) cannot be explained by an abnormal subcellular distribution. In summary, NGF stimulates phosphorylation of SH2-B on serines/threonines by kinases downstream of MEK, which may be important for NGF-mediated neuronal differentiation and survival. PMID- 10473610 TI - GTP hydrolysis links initiation and termination of nuclear import on the nucleoporin nup358. AB - Binding of GTP-bound Ran (RanGTP) to karyopherin beta1 (Kapbeta1) releases import cargo into the nucleus. Using an ultrastructural, biochemical, and functional approach, we have studied the mechanism by which Kapbeta1.RanGTP is recycled at the nuclear pore complex for repeated rounds of import. In vitro, Kapbeta1 bound to the RanBP1-homologous (RBH) domains of Nup358 in the presence of either RanGTP or RanGDP, forming trimeric complexes. The Kapbeta1.RanGTP. RBH complex resisted dissociation by RanBP1 and GTP hydrolysis by Ran GTPase activating protein 1. Ran dependent binding of gold-conjugated Kapbeta1 to the cytoplasmic fibers of the nuclear pore complex in digitonin-permeabilized cells and RanBP1 competition confirmed the in vitro binding data. Interaction of karyopherin alpha and a classical nuclear localization sequence peptide with the Kapbeta1.RanGTP.RBH complex stimulated GTP hydrolysis by Ran GTPase activating protein 1 both in vitro and in permeabilized cells. This GTP hydrolysis was required for reinitiation of import of a nuclear localization sequence-bearing substrate in permeabilized cells. These data suggest that GTP hydrolysis on the RBH domains of Nup358 couples the termination of one cycle of nuclear import with the initiation of the next. PMID- 10473611 TI - Specific activation of the p38 mitogen-activated protein kinase signaling pathway and induction of neurite outgrowth in PC12 cells by bone morphogenetic protein-2. AB - Bone morphogenetic protein (BMP)-2 has the capacity to induce the neuronal differentiation of PC12 cells. Unlike nerve growth factor, however, BMP-2 failed to induce the activation of the 41-/43-kDa mitogen-activated protein (MAP) kinase pathway in these cells. In contrast, BMP-2 characteristically induced the sustained activation of the p38 MAP kinase pathway. Pretreatment of PC12 cells with SB203580 inhibited the BMP-2-induced neurite outgrowth formation in a dose dependent manner; this inhibition coincided well with the ability of SB203580 to inihibit the BMP-2-induced activation of the p38 MAP kinase pathway. Overexpression in PC12 cells of wild-type MAP kinase kinase (MKK)-6 enhanced the BMP-2-induced activation of p38 MAP kinase, whose activation correlated well with the ability of these cells to induce neurite outgrowth in response to BMP-2. Transient expression of kinase-negative forms of MKK3/6 inhibited the formation of neurite outgrowth in response to BMP-2. Furthermore, expression of constitutively active forms of MKK3/6 induced neurite outgrowth without BMP-2 stimulation, and SB203580 inhibited this induction. These results clearly indicate that activation of the p38 MAP kinase pathway is necessary for BMP-2 induced neuronal differentiation of PC12 cells. Our results also suggest that activation of the p38 MAP kinase pathway alone can induce the neuronal differentiation of PC12 cells. PMID- 10473612 TI - Tenascin-R is a functional modulator of sodium channel beta subunits. AB - Voltage-gated sodium channels isolated from mammalian brain are composed of alpha, beta1, and beta2 subunits. The alpha subunit forms the ion conducting pore of the channel, whereas the beta1 and beta2 subunits modulate channel function, as well as channel plasma membrane expression levels. beta1 and beta2 each contain a single, extracellular Ig-like domain with structural similarity to the neural cell adhesion molecule (CAM), myelin Po. beta2 contains strong amino acid homology to the third Ig domain and to the juxtamembrane region of F3/contactin. Many CAMs of the Ig superfamily have been shown to interact with extracellular matrix molecules. We hypothesized that beta2 may interact with tenascin-R (TN-R), an extracellular matrix molecule that is secreted by oligodendrocytes during myelination and that binds F3-contactin. We show here that cells expressing sodium channel beta1 or beta2 subunits are functionally modulated by TN-R. Transfected cells stably expressing beta1 or beta2 subunits initially recognized and then were repelled from TN-R substrates. The cysteine-rich amino-terminal domain of TN-R expressed as a recombinant peptide, termed EGF-L, appears to be responsible for the repellent effect on beta subunit-expressing cells. The epidermal growth factor-like repeats and fibronectin-like repeats 6-8 are most effective in the initial adhesion of beta subunit-expressing cells. Application of EGF-L to alphaIIAbeta1beta2 channels expressed in Xenopus oocytes potentiated expressed sodium currents without significantly altering current time course or the voltage dependence of current activation or inactivation. Thus, sodium channel beta subunits appear to function as CAMs, and TN-R may be an important regulator of sodium channel localization and function in neurons. PMID- 10473613 TI - SNARE proteins regulate H(+)-ATPase redistribution to the apical membrane in rat renal inner medullary collecting duct cells. AB - The interaction of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins provides the necessary steps for vesicle docking fusion. In inner medullary collecting duct (IMCD) cells, acid secretion is regulated in part by exocytotic insertion and endocytotic retrieval of an H(+) ATPase to and from the apical membrane. We previously suggested a role for SNARE proteins in exocytotic insertion of proton pumps in IMCD cells. The purpose of the present study was to determine whether SNARE proteins are associated with the 31-kDa subunit of H(+)-ATPase in IMCD cells during exocytosis and to determine the effects of clostridial toxins on SNARE-mediated trafficking of H(+)-ATPase. Cell acidification induced a marked increment of H(+)-ATPase in the apical membrane. However, pretreating cells with clostridial toxins blocked the cellular translocation of the 31-kDa subunit. Immunoprecipitation of IMCD cell homogenate, using antibodies against either the 31-kDa subunit of H(+)-ATPase or vesicle associated membrane protein-2, co-immunoprecipitated N-ethylmaleimide-sensitive factor, alpha-soluble NSF attachment protein (alpha-SNAP), synaptosome-associated protein-23, syntaxin, and vesicle-associated membrane protein-2. Pretreatment with clostridial toxin resulted in reduced co-immunoprecipitation of H(+)-ATPase and syntaxin. These experiments document, for the first time, a putative docking fusion complex in IMCD cells and a physical association of the H(+)-ATPase with the complex. The sensitivity to the action of clostridial toxin indicates the docking-fusion complex is a part of the exocytotic mechanism of the proton pump. PMID- 10473614 TI - Molecular cloning, expression, and characterization of angiopoietin-related protein. angiopoietin-related protein induces endothelial cell sprouting. AB - Using degenerate polymerase chain reaction, we isolated a cDNA encoding a novel 493-amino acid protein from human and mouse adult heart cDNAs and have designated it angiopoietin-related protein-2 (ARP2). The NH(2)-terminal and COOH-terminal portions of ARP2 contain the characteristic coiled-coil domain and fibrinogen like domain that are conserved in angiopoietins. ARP2 has two consensus glycosylation sites and a highly hydrophobic region at the NH(2) terminus that is typical of a secretory signal sequence. Recombinant ARP2 expressed in COS cells is secreted and glycosylated. In human adult tissues, ARP2 mRNA is most abundant in heart, small intestine, spleen, and stomach. In rat embryos, ARP2 mRNA is most abundant in the blood vessels and skeletal muscles. Endothelial and vascular smooth muscle cells also contain ARP2 mRNA. Recombinant ARP2 protein induces sprouting in vascular endothelial cells but does not bind to the Tie1 or Tie2 receptor. These results suggest that ARP2 may exert a function on endothelial cells through autocrine or paracrine action. PMID- 10473615 TI - Using a biochemical approach to identify the primary dimerization regions in human DNA topoisomerase IIalpha. AB - Eukaryotic topoisomerase II is a nuclear enzyme essential for DNA metabolism and chromosome dynamics. The enzyme has a dimeric structure, and subunit dimerization is vital to the cellular functions and activities of the enzyme. Two biochemical approaches based on metal ion affinity chromatography and immunoprecipitation have been carried out to map the dimerization region(s) in human topoisomerase IIalpha. The results demonstrate that two regions spanning amino acids 1053-1069 and 1124-1143 are both essential for dimerization. The regions correspond to the interaction domains revealed in yeast topoisomerase II after crystallization of a central fragment of this enzyme, indicating that the overall C-terminal dimerization structure of eukaryotic topoisomerase II is conserved from yeast to human. Furthermore, linker insertion analysis has demonstrated that the two dimerization regions are located in a highly flexible part of the enzyme. Topoisomerase IIalpha mutant enzymes unable to dimerize via the C-terminal primary dimerization regions due to lack of one of the defined dimerization regions can still be forced to dimerize if DNA and an ATP analog are added to the reaction mixture. The result indicates that secondary interactions occur by ATP analog-mediated clamp closing when the subunits are brought together on DNA. PMID- 10473616 TI - Characterization of a novel cis-element that regulates Fas ligand expression in corneal endothelial cells. AB - Constitutively expressed Fas ligand in the cornea, Sertoli cell of the testes, Paneth cell of the intestines, and Clara cell of the airway protect surrounding parenchymal tissue by inducing apoptosis of Fas(+) immune cells during inflammatory reactions. Indeed, the action of corneal Fas ligand has been suggested to facilitate successful allogeneic cornea transplantation. To study the transcriptional regulation of Fas ligand in the eye, we employed an immortalized mouse corneal endothelial cell line (C3H3) that constitutively expresses Fas ligand. By deletion analysis of the mouse Fas ligand promoter, gel mobility shift assays, and site-directed mutagenesis, we found that a TCCT motif located -299 base pairs upstream from the transcriptional start site served as a major positive regulatory cis-element in C3H3 cells. In contrast, this element was not required for Fas ligand transcriptional activity in Sertoli cells and airway epithelial cells. By UV cross-linking analysis, we found that an approximately 30-kDa corneal nuclear protein binds to the Fas ligand promoter TCCT box and, thus, likely plays an important role in Fas ligand expression in corneal endothelial cells. PMID- 10473617 TI - Characterization of serine 916 as an in vivo autophosphorylation site for protein kinase D/Protein kinase Cmu. AB - Activation of the serine kinase protein kinase D (PKD)/PKCmicro is controlled by the phosphorylation of two serine residues within its activation loop via a PKC dependent signaling cascade. In this study we have identified the C-terminal serine 916 residue as an in vivo phosphorylation site within active PKD/PKCmu. An antibody that recognized PKD/PKCmu proteins specifically phosphorylated on the serine 916 residue was generated and used to show that phosphorylation of Ser-916 is induced by phorbol ester treatment of cells. Thus, the pS916 antibody is a useful tool to study the regulation of PKD/PKCmu activity in vivo. Antigen receptor ligation of T and B lymphocytes also induced phosphorylation of the serine 916 residue of PKD/PKCmu. Furthermore the regulatory FcgammaRIIB receptor, which mediates vital negative feedback signals to the B cell antigen receptor complex, inhibited the antigen receptor-induced activation and serine 916 phosphorylation of PKD/PKCmu. The degree of serine 916 phosphorylation during lymphocyte activation and inhibition exactly correlated with the activation status of PKD/PKCmu. Moreover, using different mutants of PKD/PKCmu, we show that serine 916 is not trans-phosphorylated by an upstream kinase but is rather an autophosphorylation event that occurs following activation of PKD/PKCmu. PMID- 10473618 TI - Abortive initiation by Saccharomyces cerevisiae RNA polymerase III. AB - Promoter escape can be rate-limiting for transcription by bacterial RNA polymerases and RNA polymerase II of higher eukaryotes. Formation of a productive elongation complex requires disengagement of RNA polymerase from promoter-bound eukaryotic transcription factors or bacterial sigma factors. RNA polymerase III (pol III) stably associates with the TFIIIB-DNA complex even in the absence of localized DNA unwinding associated with the open promoter complex. To explore the role that release of pol III from the TFIIIB-DNA complex plays in limiting the overall rate of transcription, we have examined the early steps of RNA synthesis. We find that, on average, only three rounds of abortive initiation precede the formation of each elongation complex and that nearly all pol III molecules escape the abortive initiation phase of transcription without significant pausing or arrest. However, when elongation is limited to 5 nucleotides, the intrinsic exoribonuclease activity of pol III cleaves 5-mer RNA at a rate considerably faster than product release or reinitiation. This cleavage also occurs in the normal process of forming a productive elongation complex. The possible role of nucleolytic retraction in disengaging pol III from TFIIIB is discussed. PMID- 10473619 TI - Molecular directionality of polysaccharide polymerization by the Pasteurella multocida hyaluronan synthase. AB - Hyaluronan (HA), a long linear polymer composed of alternating glucuronic acid and N-acetylglucosamine residues, is an essential polysaccharide in vertebrates and a putative virulence factor in certain microbes. All known HA synthases utilize UDP-sugar precursors. Previous reports describing the HA synthase enzymes from Streptococcus bacteria and mammals, however, did not agree on the molecular directionality of polymer elongation. We show here that a HA synthase, PmHAS, from Gram-negative P. multocida bacteria polymerizes the HA chain by the addition of sugar units to the nonreducing terminus. Recombinant PmHAS will elongate exogenous HA oligosaccharide acceptors to form long polymers in vitro; thus far no other HA synthase has displayed this capability. The directionality of synthesis was established definitively by testing the ability of PmHAS to elongate defined oligosaccharide derivatives. Analysis of the initial stages of synthesis demonstrated that PmHAS added single monosaccharide units sequentially. Apparently the fidelity of the individual sugar transfer reactions is sufficient to generate the authentic repeating structure of HA. Therefore, simultaneous addition of disaccharide block units is not required as hypothesized in some recent models of polysaccharide biosynthesis. PmHAS appears distinct from other known HA synthases based on differences in sequence, topology in the membrane, and putative reaction mechanism. PMID- 10473620 TI - Activation of the protein kinase ERK5/BMK1 by receptor tyrosine kinases. Identification and characterization of a signaling pathway to the nucleus. AB - ERK5 (also known as BMK1), a member of the mitogen-activated protein kinase (MAPK) superfamily, was known to be activated strongly by oxidant and osmotic stresses. Here we have found that ERK5 is strongly activated by epidermal growth factor and nerve growth factor, whose receptors are tyrosine kinases. The activation of ERK5 was inhibited by expression of dominant-negative Ras and induced by expression of active Ras in PC12 cells, indicating a requirement for Ras in ERK5 activation. The epidermal growth factor-induced activation of ERK5 was found to be inhibited by PD98059 and U0126 inhibitors, which were previously thought to act specifically on classical MAPK kinase (also known as MEK1) and readily reversed by CL100 and MKP-3 dual-specificity phosphatases for which classical MAPKs were previously shown to serve as preferred substrates. The reporter assays demonstrated that the serum-induced enhancement of transcription from serum response element was significantly inhibited by expression of a dominant-negative form of MEK5, which was a direct and specific activator for ERK5 and that transcription from serum response element mediated by the Ets domain transcription factor Sap1a, but not by Elk1, was stimulated by coexpression of ERK5 and active MEK5. In addition, Sap1a was shown to be phosphorylated by ERK5 in vitro and by the activation of the ERK5 pathway in cells. Moreover, the serum-induced c-Fos expression was markedly inhibited by expression of dominant-negative MEK5. These results reveal a novel signaling pathway to the nucleus mediated by ERK5 that functions downstream of receptor tyrosine kinases to induce immediate early genes, in parallel with the classical MAPK cascade. PMID- 10473621 TI - Ribonuclease III processing of coaxially stacked RNA helices. AB - The RNase III family of endoribonucleases participates in maturation and decay of cellular and viral transcripts by processing of double-stranded RNA. RNase III degradation is inherent to most antisense RNA-regulated gene systems in Escherichia coli. In the hok/sok system from plasmid R1, Sok antisense RNA targets the hok mRNA for RNase III-mediated degradation. An intermediate in the pairing reaction between Sok RNA and hok mRNA forms a three-way junction. A complex between a chimeric antisense RNA and hok mRNA that mimics the three-way junction was cleaved by RNase III both in vivo and in vitro. Footprinting using E117A RNase III binding to partially complementary RNAs showed protection of the 13 base pairs of interstrand duplex and of the bottom part of the transcriptional terminator hairpin of the antisense RNA. This suggests that the 13 base pairs of RNA duplex are coaxially stacked on the antisense RNA terminator stem-loop and that each stem forms a monomer half-site, allowing symmetrical binding of the RNase III dimer. This processing scheme shows an unanticipated diversity in RNase III substrates and may have a more general implication for RNA metabolism. PMID- 10473622 TI - SH2-kinase linker mutations release Hck tyrosine kinase and transforming activities in Rat-2 fibroblasts. AB - Biochemical and structural studies of Src and related kinases demonstrate that two intramolecular interactions suppress kinase activity. These interactions involve binding of the SH2 domain to a phosphotyrosine residue in the C-terminal tail and association of the SH3 domain with a polyproline type II helix formed by amino acids linking the SH2 and kinase domains. Recent studies have shown that high affinity interaction of the SH3 domain of Hck with the human immunodeficiency virus type I Nef protein activates Hck tyrosine kinase and biological activities, suggesting a mechanism that involves disruption of the SH3 linker interaction. To test the role of this interaction in the regulation of Hck kinase activity in living cells, we substituted alanines for prolines 225 and 228 in the linker region and observed that the resulting mutant (Hck-2PA) demonstrated strong transforming activity in a Rat-2 fibroblast focus-forming assay. Hck-2PA also exhibited elevated tyrosine kinase activity in terms of autophosphorylation, endogenous substrate phosphorylation, and in an in vitro kinase assay. The transforming and kinase activities of Hck-2PA were remarkably similar to those observed with a Hck mutant activated by Phe substitution of the conserved tail Tyr residue and with wild-type Hck following co-expression with human immunodeficiency virus Nef. Introduction of the 2PA and tail mutations into a single Hck expression construct did not increase kinase or transforming activity relative to the individual mutations. These data provide new evidence that SH3-linker interaction may represent the dominant mechanism controlling Hck tyrosine kinase activity in vivo. PMID- 10473623 TI - The role of FAST-1 and Smads in transcriptional regulation by activin during early Xenopus embryogenesis. AB - Smads are signal transducers for the transforming growth factor-beta superfamily of factors. In early Xenopus embryos, the transforming growth factor-beta member activin induces the gene Mix.2 by stimulating the formation of a multiprotein complex, activin-responsive factor (ARF). This complex contains Smad2 or Smad3, Smad4, and a novel forkhead transcription factor, FAST-1, and binds to an enhancer (activin-responsive element; ARE) that confers activin regulation of Mix.2 transcription. Both FAST-1 and Smads can bind directly to the ARE; we have investigated 1) the role of FAST-1 and Smad DNA binding sites in ARF recognition of the ARE, 2) the contributions of FAST-1 and Smad binding to ARF binding in vitro and to ARE regulation in early Xenopus embryos, 3) the extent to which different Smads can replace Smad4 in regulation of the ARE. We find that ARF binds to ARE through both FAST-1 and Smad binding sites. FAST-1 recognition of the ARE is essential both for ARF binding in vitro and activin regulation in vivo. In contrast, Smad binding of ARE is unnecessary for ARF binding or activin regulation but does enhance the binding and regulatory activity of ARF. Also, Smad3 can partially substitute for Smad4 in the regulation of the ARE. These observations elucidate how broadly expressed signal transducers (Smads) regulate a developmentally specific transcriptional response in conjunction with a temporally restricted transcription factor, FAST-1. PMID- 10473624 TI - CCAAT/enhancer-binding proteins regulate expression of the human steroidogenic acute regulatory protein (StAR) gene. AB - Two putative CCAAT/enhancer-binding protein (C/EBP) response elements were identified in the proximal promoter of the human steroidogenic acute regulatory protein (StAR) gene, which encodes a key protein-regulating steroid hormone synthesis. Expression of C/EBPalpha and -beta increased StAR promoter activity in COS-1 and HepG2 cells. Cotransfection of C/EBPalpha or -beta and steroidogenic factor 1, a transcription factor required for cAMP regulation of StAR expression, into COS-1 augmented 8-bromoadenosine 3':5'-cyclic monophosphate (8-Br-cAMP) stimulated promoter activity. When the putative C/EBP response elements were mutated, individually or together, a pronounced decline in basal StAR promoter activity in human granulosa-lutein cells resulted, but the fold stimulation of promoter activity by 8-Br-cAMP was unaffected. Recombinant C/EBPalpha and -beta bound to the two identified sequences but not the mutated elements. Human granulosa-lutein cell nuclear extracts also bound these elements but not the mutated sequences. An antibody to C/EBPbeta, but not C/EBPalpha, supershifted the nuclear protein complex associated with the more distal element. The complex formed by nuclear extracts with the proximal element was not supershifted by either antibody. Western blot analysis revealed the presence of C/EBPalpha and C/EBPbeta in human granulosa-lutein cell nuclear extracts. C/EBPbeta levels were up-regulated 3-fold by 8-Br-cAMP treatment. Our studies demonstrate a role for C/EBPbeta as well as yet to be identified proteins, which can bind to C/EBP response elements, in the regulation of StAR gene expression and suggest a mechanism by which C/EBPbeta participates in the cAMP regulation of StAR gene transcription. PMID- 10473626 TI - Biochemical characterization of the epithelial dystroglycan complex. AB - Dystroglycan is a widely expressed extracellular matrix receptor that plays a critical role in basement membrane formation, epithelial development, and synaptogenesis. Dystroglycan was originally characterized in skeletal muscle as an integral component of the dystrophin glycoprotein complex, which is critical for muscle cell viability. Although the dystroglycan complex has been well characterized in skeletal muscle, there is little information on the structural composition of the dystroglycan complex outside skeletal muscle. Here we have biochemically characterized the dystroglycan complex in lung and kidney. We demonstrate that the presence of sarcoglycans and sarcospan in lung reflects association with dystroglycan in the smooth muscle. The smooth muscle dystroglycan complex in lung, composed of dystroglycan, dystrophin/utrophin, beta , delta-, epsilon-sarcoglycan, and sarcospan, can be biochemically separated from epithelial dystroglycan, which is not associated with any of the known sarcoglycans or sarcospan. Similarly, dystroglycan in kidney epithelial cells is not associated with any of the sarcoglycans or sarcospan. Thus, our data demonstrate that there are distinct dystroglycan complexes in non-skeletal muscle organs as follows: one from smooth muscle, which is associated with sarcoglycans forming a similar complex as in skeletal muscle, and one from epithelial cells. PMID- 10473625 TI - Regulation of the Dlx3 homeobox gene upon differentiation of mouse keratinocytes. AB - The Distal-less Dlx3 homeodomain gene is expressed in terminally differentiated murine epidermal cells, and there is evidence to support an essential role as a transcriptional regulator of the terminal differentiation process in these cells. In an attempt to determine the factors that induce Dlx3 gene expression, we have cloned the 1.2-kilobase pair proximal region of murine gene and analyzed its cis regulatory elements and potential trans-acting factors. The proximal region of the Dlx3 gene has a canonical TATA box and CCAAT box, and the transcription start site was located 205 base pairs upstream from the initiation of translation site. Serial deletion analysis showed that the region between -84 and -34 confers the maximal promoter activity both in undifferentiated and differentiated primary mouse keratinocytes. Gel retardation assays and mutational analysis demonstrated that the transcriptional regulator NF-Y (also referred to as CBF) binds to a CCAAT box motif within this region and is responsible for the majority of the Dlx3 promoter activity. In addition, an Sp1-binding site was located immediately upstream of transcription start site that acts as a positive regulatory element of the Dlx3 promoter, independent of the CCAAT box motif. Importantly, elements residing between +30 to +60 of the Dlx3 gene are responsible for the Ca(2+) dependent induction of Dlx3 during keratinocyte differentiation. PMID- 10473627 TI - L-Glutamic acid gamma-monohydroxamate. A potentiator of vanadium-evoked glucose metabolism in vitro and in vivo. AB - We report that the vanadium ligand L-Glu(gamma)HXM potentiates the capacity of free vanadium ions to activate glucose uptake and glucose metabolism in rat adipocytes in vitro (by 4-5-fold) and to lower blood glucose levels in hyperglycemic rats in vivo (by 5-7-fold). A molar ratio of two L-Glu(gamma)HXM molecules to one vanadium ion was most effective. Unlike other vanadium ligands that potentiate the insulinomimetic actions of vanadium, L-Glu(gamma)HXM partially activated lipogenesis in rat adipocytes in the absence of exogenous vanadium. This effect was not manifested by D-Glu(gamma)HXM. At 10-20 microM L Glu(gamma)HXM, lipogenesis was activated 9-21%. This effect was approximately 9 fold higher (140 +/- 15% of maximal insulin response) in adipocytes derived from rats that had been treated with vanadium for several days. Titration of vanadium(IV) with L-Glu(gamma)HXM led to a rapid decrease in the absorbance of vanadium(IV) at 765 nm, and (51)V NMR spectroscopy revealed that the chemical shift of vanadium(IV) at -490 ppm disappeared with the appearance of a signal characteristic to vanadium(V) (-530 ppm) upon adding one equivalent of L Glu(gamma)HXM. In summary, L-Glu(gamma)HXM is highly active in potentiating vanadium-activated glucose metabolism in vitro and in vivo and facilitating glucose metabolism in rat adipocytes in the absence of exogenous vanadium probably through conversion of trace intracellular vanadium into an active insulinomimetic compound. We propose that the active species is either a 1:1 or 2:1 L-Glu(gamma)HXM vanadium complex in which the endogenous vanadium(IV) has been altered to vanadium(V). Finally we demonstrate that L-Glu(gamma)HXM- and L Glu(gamma)HXM.vanadium-evoked lipogenesis is arrested by wortmannin and that activation of glucose uptake in rat adipocytes is because of enhanced translocation of GLUT4 from low density microsomes to the plasma membrane. PMID- 10473628 TI - The role of the membrane-spanning domain sequence in glycoprotein-mediated membrane fusion. AB - The role of glycoprotein membrane-spanning domains in the process of membrane fusion is poorly understood. It has been demonstrated that replacing all or part of the membrane-spanning domain of a viral fusion protein with sequences that encode signals for glycosylphosphatidylinositol linkage attachment abrogates membrane fusion activity. It has been suggested, however, that the actual amino acid sequence of the membrane-spanning domain is not critical for the activity of viral fusion proteins. We have examined the function of Moloney murine leukemia virus envelope proteins with substitutions in the membrane-spanning domain. Envelope proteins bearing substitutions for proline 617 are processed and incorporated into virus particles normally and bind to the viral receptor. However, they possess greatly reduced or undetectable capacities for the promotion of membrane fusion and infectious virus particle formation. Our results imply a direct role for the residues in the membrane-spanning domain of the murine leukemia virus envelope protein in membrane fusion and its regulation. They also support the thesis that membrane-spanning domains possess a sequence dependent function in other protein-mediated membrane fusion events. PMID- 10473629 TI - Division of labor among the alpha6beta4 integrin, beta1 integrins, and an E3 laminin receptor to signal morphogenesis and beta-casein expression in mammary epithelial cells. AB - Contact of cultured mammary epithelial cells with the basement membrane protein laminin induces multiple responses, including cell shape changes, growth arrest, and, in the presence of prolactin, transcription of the milk protein beta-casein. We sought to identify the specific laminin receptor(s) mediating the multiple cell responses to laminin. Using assays with clonal mammary epithelial cells, we reveal distinct functions for the alpha6beta4 integrin, beta1 integrins, and an E3 laminin receptor. Signals from laminin for beta-casein expression were inhibited in the presence of function-blocking antibodies against both the alpha6 and beta1 integrin subunits and by the laminin E3 fragment. The alpha6-blocking antibody perturbed signals mediated by the alpha6beta4 integrin, and the beta1 blocking antibody perturbed signals mediated by another integrin, the alpha subunit(s) of which remains to be determined. Neither alpha6- nor beta1-blocking antibodies perturbed the cell shape changes resulting from cell exposure to laminin. However, the E3 laminin fragment and heparin both inhibited cell shape changes induced by laminin, thereby implicating an E3 laminin receptor in this function. These results elucidate the multiplicity of cell-extracellular matrix interactions required to integrate cell structure and signaling and ultimately permit normal cell function. PMID- 10473630 TI - A novel Ras-interacting protein required for chemotaxis and cyclic adenosine monophosphate signal relay in Dictyostelium. AB - We have identified a novel Ras-interacting protein from Dictyostelium, RIP3, whose function is required for both chemotaxis and the synthesis and relay of the cyclic AMP (cAMP) chemoattractant signal. rip3 null cells are unable to aggregate and lack receptor activation of adenylyl cyclase but are able, in response to cAMP, to induce aggregation-stage, postaggregative, and cell-type-specific gene expression in suspension culture. In addition, rip3 null cells are unable to properly polarize in a cAMP gradient and chemotaxis is highly impaired. We demonstrate that cAMP stimulation of guanylyl cyclase, which is required for chemotaxis, is reduced approximately 60% in rip3 null cells. This reduced activation of guanylyl cyclase may account, in part, for the defect in chemotaxis. When cells are pulsed with cAMP for 5 h to mimic the endogenous cAMP oscillations that occur in wild-type strains, the cells will form aggregates, most of which, however, arrest at the mound stage. Unlike the response seen in wild-type strains, the rip3 null cell aggregates that form under these experimental conditions are very small, which is probably due to the rip3 null cell chemotaxis defect. Many of the phenotypes of the rip3 null cell, including the inability to activate adenylyl cyclase in response to cAMP and defects in chemotaxis, are very similar to those of strains carrying a disruption of the gene encoding the putative Ras exchange factor AleA. We demonstrate that aleA null cells also exhibit a defect in cAMP-mediated activation of guanylyl cyclase similar to that of rip3 null cells. A double-knockout mutant (rip3/aleA null cells) exhibits a further reduction in receptor activation of guanylyl cyclase, and these cells display almost no cell polarization or movement in cAMP gradients. As RIP3 preferentially interacts with an activated form of the Dictyostelium Ras protein RasG, which itself is important for cell movement, we propose that RIP3 and AleA are components of a Ras-regulated pathway involved in integrating chemotaxis and signal relay pathways that are essential for aggregation. PMID- 10473631 TI - Insulin-induced stimulation of Na+,K(+)-ATPase activity in kidney proximal tubule cells depends on phosphorylation of the alpha-subunit at Tyr-10. AB - Phosphorylation of the alpha-subunit of Na+,K(+)-ATPase plays an important role in the regulation of this pump. Recent studies suggest that insulin, known to increase solute and fluid reabsorption in mammalian proximal convoluted tubule (PCT), is stimulating Na+,K(+)-ATPase activity through the tyrosine phosphorylation process. This study was therefore undertaken to evaluate the role of tyrosine phosphorylation of the Na+,K(+)-ATPase alpha-subunit in the action of insulin. In rat PCT, insulin and orthovanadate (a tyrosine phosphatase inhibitor) increased tyrosine phosphorylation level of the alpha-subunit more than twofold. Their effects were not additive, suggesting a common mechanism of action. Insulin induced tyrosine phosphorylation was prevented by genistein, a tyrosine kinase inhibitor. The site of tyrosine phosphorylation was identified on Tyr-10 by controlled trypsinolysis in rat PCTs and by site-directed mutagenesis in opossum kidney cells transfected with rat alpha-subunit. The functional relevance of Tyr 10 phosphorylation was assessed by 1) the abolition of insulin-induced stimulation of the ouabain-sensitive (86)Rb uptake in opossum kidney cells expressing mutant rat alpha1-subunits wherein tyrosine was replaced by alanine or glutamine; and 2) the similarity of the time course and dose dependency of the insulin-induced increase in ouabain-sensitive (86)Rb uptake and tyrosine phosphorylation. These findings indicate that phosphorylation of the Na+,K(+) ATPase alpha-subunit at Tyr-10 likely participates in the physiological control of sodium reabsorption in PCT. PMID- 10473632 TI - Stimulation of beta1-integrin function by epidermal growth factor and heregulin beta has distinct requirements for erbB2 but a similar dependence on phosphoinositide 3-OH kinase. AB - Integrins and growth factor receptors are important participants in cellular adhesion and migration. The EGF receptor (EGFR) family of tyrosine kinases and the beta1-integrin adhesion receptors are of particular interest, given the implication for their involvement in the initiation and progression of tumorigenesis. We used adhesion and chemotaxis assays to further elucidate the relationship between these two families of transmembrane signaling molecules. Specifically, we examined integrin-mediated adhesive and migratory characteristics of the metastatic breast carcinoma cell line MDA-MB-435 in response to stimulation with growth factors that bind to and activate the EGFR or erbB3 in these cells. Although ligand engagement of the EGFR stimulated modest beta1-dependent increases in cell adhesion and motility, heregulin-beta (HRGbeta) binding to the erbB3 receptor initiated rapid and potent induction of breast carcinoma cell adhesion and migration and required dimerization of erbB3 with erbB2. Pharmacologic inhibitors of phosphoinositide 3-OH kinase (PI 3-K) or transient expression of dominant negative forms of PI 3-K inhibited both EGF- and HRGbeta-mediated adhesion and potently blocked HRGbeta- and EGF-induced cell motility. Our results illustrate the critical role of PI 3-K activity in signaling pathways initiated by the EGFR or erbB3 to up-regulate beta1-integrin function. PMID- 10473633 TI - Import into and degradation of cytosolic proteins by isolated yeast vacuoles. AB - In eukaryotic cells, both lysosomal and nonlysosomal pathways are involved in degradation of cytosolic proteins. The physiological condition of the cell often determines the degradation pathway of a specific protein. In this article, we show that cytosolic proteins can be taken up and degraded by isolated Saccharomyces cerevisiae vacuoles. After starvation of the cells, protein uptake increases. Uptake and degradation are temperature dependent and show biphasic kinetics. Vacuolar protein import is dependent on cytosolic heat shock proteins of the hsp70 family and on protease-sensitive component(s) on the outer surface of vacuoles. Degradation of the imported cytosolic proteins depends on a functional vacuolar ATPase. We show that the cytosolic isoform of yeast glyceraldehyde-3-phosphate dehydrogenase is degraded via this pathway. This import and degradation pathway is reminiscent of the protein transport pathway from the cytosol to lysosomes of mammalian cells. PMID- 10473634 TI - Early endosomes are required for major histocompatiblity complex class II transport to peptide-loading compartments. AB - Antigen presentation to CD4(+) T lymphocytes requires transport of newly synthesized major histocompatibility complex (MHC) class II molecules to the endocytic pathway, where peptide loading occurs. This step is mediated by a signal located in the cytoplasmic tail of the MHC class II-associated Ii chain, which directs the MHC class II-Ii complexes from the trans-Golgi network (TGN) to endosomes. The subcellular machinery responsible for the specific targeting of MHC class II molecules to the endocytic pathway, as well as the first compartments these molecules enter after exit from the TGN, remain unclear. We have designed an original experimental approach to selectively analyze this step of MHC class II transport. Newly synthesized MHC class II molecules were caused to accumulate in the Golgi apparatus and TGN by incubating the cells at 19 degrees C, and early endosomes were functionally inactivated by in vivo cross linking of transferrin (Tf) receptor-containing endosomes using Tf-HRP complexes and the HRP-insoluble substrate diaminobenzidine. Inactivation of Tf-containing endosomes caused a marked delay in Ii chain degradation, peptide loading, and MHC class II transport to the cell surface. Thus, early endosomes appear to be required for delivery of MHC class II molecules to the endocytic pathway. Under cross-linking conditions, most alphabetaIi complexes accumulated in tubules and vesicles devoid of gamma-adaptin and/or mannose-6-phosphate receptor, suggesting an AP1-independent pathway for the delivery of newly synthesized MHC class II molecules from the TGN to endosomes. PMID- 10473635 TI - Rad18 is required for DNA repair and checkpoint responses in fission yeast. AB - To survive damage to the genome, cells must respond by activating both DNA repair and checkpoint responses. Using genetic screens in the fission yeast Schizosaccharomyces pombe, we recently isolated new genes required for DNA damage checkpoint control. We show here that one of these strains defines a new allele of the previously described rad18 gene, rad18-74. rad18 is an essential gene, even in the absence of extrinsic DNA damage. It encodes a conserved protein related to the structural maintenance of chromosomes proteins. Point mutations in rad18 lead to defective DNA repair pathways responding to both UV-induced lesions and, as we show here, double-stranded breaks. Furthermore, rad18p is required to maintain cell cycle arrest in the presence of DNA damage, and failure of this leads to highly aberrant mitoses. A gene encoding a BRCT-containing protein, brc1, was isolated as an allele-specific high-copy suppressor of rad18-74. brc1 is required for mitotic fidelity and for cellular viability in strains with rad18 mutations but is not essential for DNA damage responses. Mutations in rad18 and brc1 are synthetically lethal with a topoisomerase II mutant (top2-191), indicating that these proteins play a role in chromatin organization. These studies show a role for chromatin organization in the maintenance or activation of responses to DNA damage. PMID- 10473636 TI - Neurite extension occurs in the absence of regulated exocytosis in PC12 subclones. AB - We have investigated the process leading to differentiation of PC12 cells. This process is known to include extension of neurites and changes in the expression of subsets of proteins involved in cytoskeletal rearrangements or in neurosecretion. To this aim, we have studied a PC12 clone (trk-PC12) stably transfected with the nerve growth factor receptor TrkA. These cells are able to undergo both spontaneous and neurotrophin-induced morphological differentiation. However, both undifferentiated and nerve growth factor-differentiated trk-PC12 cells appear to be completely defective in the expression of proteins of the secretory apparatus, including proteins of synaptic vesicles and large dense-core granules, neurotransmitter transporters, and neurotransmitter-synthesizing enzymes. These results indicate that neurite extension can occur independently of the presence of the neurosecretory machinery, including the proteins that constitute the fusion machine, suggesting the existence of differential activation pathways for the two processes during neuronal differentiation. These findings have been confirmed in independent clones obtained from PC12-27, a previously characterized PC12 variant clone globally incompetent for regulated secretion. In contrast, the integrity of the Rab cycle appears to be necessary for neurite extension, because antisense oligonucleotides against the neurospecific isoform of Rab-guanosine diphosphate-dissociation inhibitor significantly interfere with process formation. PMID- 10473637 TI - The fibrinogen globe of tenascin-C promotes basic fibroblast growth factor induced endothelial cell elongation. AB - To investigate the potential role of tenascin-C (TN-C) on endothelial sprouting we used bovine aortic endothelial cells (BAECs) as an in vitro model of angiogenesis. We found that TN-C is specifically expressed by sprouting and cord forming BAECs but not by nonsprouting BAECs. To test whether TN-C alone or in combination with basic fibroblast growth factor (bFGF) can enhance endothelial sprouting or cord formation, we used BAECs that normally do not sprout and, fittingly, do not express TN-C. In the presence of bFGF, exogenous TN-C but not fibronectin induced an elongated phenotype in nonsprouting BAECs. This phenotype was due to altered actin cytoskeleton organization. The fibrinogen globe of the TN-C molecule was the active domain promoting the elongated phenotype in response to bFGF. Furthermore, we found that the fibrinogen globe was responsible for reduced cell adhesion of BAECs on TN-C substrates. We conclude that bFGF stimulated endothelial cells can be switched to a sprouting phenotype by the decreased adhesive strength of TN-C, mediated by the fibrinogen globe. PMID- 10473638 TI - Microtubule-dependent recruitment of Staufen-green fluorescent protein into large RNA-containing granules and subsequent dendritic transport in living hippocampal neurons. AB - Dendritic mRNA transport and local translation at individual potentiated synapses may represent an elegant way to form synaptic memory. Recently, we characterized Staufen, a double-stranded RNA-binding protein, in rat hippocampal neurons and showed its presence in large RNA-containing granules, which colocalize with microtubules in dendrites. In this paper, we transiently transfect hippocampal neurons with human Staufen-green fluorescent protein (GFP) and find fluorescent granules in the somatodendritic domain of these cells. Human Stau-GFP granules show the same cellular distribution and size and also contain RNA, as already shown for the endogenous Stau particles. In time-lapse videomicroscopy, we show the bidirectional movement of these Staufen-GFP-labeled granules from the cell body into dendrites and vice versa. The average speed of these particles was 6.4 microm/min with a maximum velocity of 24. 3 microm/min. Moreover, we demonstrate that the observed assembly into granules and their subsequent dendritic movement is microtubule dependent. Taken together, we have characterized a novel, nonvesicular, microtubule-dependent transport pathway involving RNA-containing granules with Staufen as a core component. This is the first demonstration in living neurons of movement of an essential protein constituent of the mRNA transport machinery. PMID- 10473639 TI - Biparental inheritance of gamma-tubulin during human fertilization: molecular reconstitution of functional zygotic centrosomes in inseminated human oocytes and in cell-free extracts nucleated by human sperm. AB - Human sperm centrosome reconstitution and the parental contributions to the zygotic centrosome are examined in mammalian zygotes and after exposure of spermatozoa to Xenopus laevis cell-free extracts. The presence and inheritance of the conserved centrosomal constituents gamma-tubulin, centrin, and MPM-2 (which detects phosphorylated epitopes) are traced, as is the sperm microtubule nucleating capability on reconstituted centrosomes. gamma-Tubulin is biparentally inherited in humans (maternal >> than paternal): Western blots detect the presence of paternal gamma-tubulin. Recruitment of maternal gamma-tubulin to the sperm centrosome occurs after sperm incorporation in vivo or exposure to cell free extract, especially after sperm "priming" induced by disulfide bond reduction. Centrin is found in the proximal sperm centrosomal region, demonstrates expected calcium sensitivity, but appears absent from the zygotic centrosome after sperm incorporation or exposure to extracts. Sperm centrosome phosphorylation is detected after exposure of primed sperm to egg extracts as well as during the early stages of sperm incorporation after fertilization. Finally, centrosome reconstitution in cell-free extracts permits sperm aster microtubule assembly in vitro. Collectively, these results support a model of a blended zygotic centrosome composed of maternal constituents attracted to an introduced paternal template after insemination. PMID- 10473641 TI - Functional dissection and hierarchy of tubulin-folding cofactor homologues in fission yeast. AB - We describe the isolation of fission yeast homologues of tubulin-folding cofactors B (Alp11) and E (Alp21), which are essential for cell viability and the maintenance of microtubules. Alp11(B) contains the glycine-rich motif (the CLIP 170 domain) involved in microtubular functions, whereas, unlike mammalian cofactor E, Alp21(E) does not. Both mammalian and yeast cofactor E, however, do contain leucine-rich repeats. Immunoprecipitation analysis shows that Alp11(B) interacts with both alpha-tubulin and Alp21(E), but not with the cofactor D homologue Alp1, whereas Alp21(E) also interacts with Alp1(D). The cellular amount of alpha-tubulin is decreased in both alp1 and alp11 mutants. Overproduction of Alp11(B) results in cell lethality and the disappearance of microtubules, which is rescued by co-overproduction of alpha-tubulin. Both full-length Alp11(B) and the C-terminal third containing the CLIP-170 domain localize in the cytoplasm, and this domain is required for efficient binding to alpha-tubulin. Deletion of alp11 is suppressed by multicopy plasmids containing either alp21(+) or alp1(+), whereas alp21 deletion is rescued by overexpression of alp1(+) but not alp11(+). Finally, the alp1 mutant is not complemented by either alp11(+) or alp21(+). The results suggest that cofactors operate in a linear pathway (Alp11(B)-Alp21(E) Alp1(D)), each with distinct roles. PMID- 10473642 TI - A nuclear protein involved in apoptotic-like DNA degradation in Stylonychia: implications for similar mechanisms in differentiating and starved cells. AB - Ciliates are unicellular eukaryotic organisms containing two types of nuclei: macronuclei and micronuclei. After the sexual pathway takes place, a new macronucleus is formed from a zygote nucleus, whereas the old macronucleus is degraded and resorbed. In the course of macronuclear differentiation, polytene chromosomes are synthesized that become degraded again after some hours. Most of the DNA is eliminated, and the remaining DNA is fragmented into small DNA molecules that are amplified to a high copy number in the new macronucleus. The protein Pdd1p (programmed DNA degradation protein 1) from Tetrahymena has been shown to be present in macronuclear anlagen in the DNA degradation stage and also in the old macronuclei, which are resorbed during the formation of the new macronucleus. In this study the identification and localization of a Pdd1p homologous protein in Stylonychia (Spdd1p) is described. Spdd1p is localized in the precursor nuclei in the DNA elimination stage and in the old macronuclei during their degradation, but also in macronuclei and micronuclei of starved cells. In all of these nuclei, apoptotic-like DNA breakdown was detected. These data suggest that Spdd1p is a general factor involved in programmed DNA degradation in Stylonychia. PMID- 10473640 TI - A p90(rsk) mutant constitutively interacting with MAP kinase uncouples MAP kinase from p34(cdc2)/cyclin B activation in Xenopus oocytes. AB - The efficient activation of p90(rsk) by MAP kinase requires their interaction through a docking site located at the C-terminal end of p90(rsk). The MAP kinase p42(mpk1) can associate with p90(rsk) in G(2)-arrested but not in mature Xenopus oocytes. In contrast, an N-terminally truncated p90(rsk) mutant named D2 constitutively interacts with p42(mpk1). In this report we show that expression of D2 inhibits Xenopus oocyte maturation. The inhibition requires the p42(mpk1) docking site. D2 expression uncouples the activation of p42(mpk1) and p34(cdc2)/cyclin B in response to progesterone but does not prevent signaling through p90(rsk). Instead, D2 interferes with a p42(mpk1)-triggered pathway, which regulates the phosphorylation and activation of Plx1, a potential activator of the Cdc25 phosphatase. This new pathway that links the activation of p42(mpk1) and Plx1 during oocyte maturation is independent of p34(cdc2)/cyclin B activity but requires protein synthesis. Using D2, we also provide evidence that the sustained activation of p42(mpk1) can trigger nuclear migration in oocytes. Our results indicate that D2 is a useful tool to study MAP kinase function(s) during oocyte maturation. Truncated substrates such as D2, which constitutively interact with MAP kinases, may also be helpful to study signal transduction by MAP kinases in other cellular processes. PMID- 10473643 TI - A role for the GSG domain in localizing Sam68 to novel nuclear structures in cancer cell lines. AB - The GSG (GRP33, Sam68, GLD-1) domain is a protein module found in an expanding family of RNA-binding proteins. The numerous missense mutations identified genetically in the GSG domain support its physiological role. Although the exact function of the GSG domain is not known, it has been shown to be required for RNA binding and oligomerization. Here it is shown that the Sam68 GSG domain plays a role in protein localization. We show that Sam68 concentrates into novel nuclear structures that are predominantly found in transformed cells. These Sam68 nuclear bodies (SNBs) are distinct from coiled bodies, gems, and promyelocytic nuclear bodies. Electron microscopic studies show that SNBs are distinct structures that are enriched in phosphorus and nitrogen, indicating the presence of nucleic acids. A GFP-Sam68 fusion protein had a similar localization as endogenous Sam68 in HeLa cells, diffusely nuclear with two to five SNBs. Two other GSG proteins, the Sam68-like mammalian proteins SLM-1 and SLM-2, colocalized with endogenous Sam68 in SNBs. Different GSG domain missense mutations were investigated for Sam68 protein localization. Six separate classes of cellular patterns were obtained, including exclusive SNB localization and association with microtubules. These findings demonstrate that the GSG domain is involved in protein localization and define a new compartment for Sam68, SLM-1, and SLM-2 in cancer cell lines. PMID- 10473644 TI - Fusion of endosomes involved in synaptic vesicle recycling. AB - Recycling of vesicles of the regulated secretory pathway presumably involves passage through an early endosomal compartment as an intermediate step. To learn more about the involvement of endosomes in the recycling of synaptic and secretory vesicles we studied in vitro fusion of early endosomes derived from pheochromocytoma (PC12) cells. Fusion was not affected by cleavage of the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins synaptobrevin and syntaxin 1 that operate at the exocytotic limb of the pathway. Furthermore, fusion was inhibited by the fast Ca(2+) chelator 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetra-acetic acid but not by the slow Ca(2+) chelator EGTA. Endosome fusion was restored by the addition of Ca(2+) with an optimum at a free Ca(2+) concentration of 0.3 x 10(-6) M. Other divalent cations did not substitute for Ca(2+). A membrane-permeant EGTA derivative caused inhibition of fusion, which was reversed by addition of Ca(2+). We conclude that the fusion of early endosomes participating in the recycling of synaptic and neurosecretory vesicles is mediated by a set of SNAREs distinct from those involved in exocytosis and requires the local release of Ca(2+) from the endosomal interior. PMID- 10473645 TI - The conserved core of a human SIR2 homologue functions in yeast silencing. AB - Silencing is a universal form of transcriptional regulation in which regions of the genome are reversibly inactivated by changes in chromatin structure. Sir2 (Silent Information Regulator) protein is unique among the silencing factors in Saccharomyces cerevisiae because it silences the rDNA as well as the silent mating-type loci and telomeres. Discovery of a gene family of Homologues of Sir Two (HSTs) in organisms from bacteria to humans suggests that SIR2's silencing mechanism might be conserved. The Sir2 and Hst proteins share a core domain, which includes two diagnostic sequence motifs of unknown function as well as four cysteines of a putative zinc finger. We demonstrate by mutational analyses that the conserved core and each of its motifs are essential for Sir2p silencing. Chimeras between Sir2p and a human Sir2 homologue (hSir2Ap) indicate that this human protein's core can substitute for that of Sir2p, implicating the core as a silencing domain. Immunofluorescence studies reveal partially disrupted localization, accounting for the yeast-human chimeras' ability to function at only a subset of Sir2p's target loci. Together, these results support a model for the involvement of distinct Sir2p-containing complexes in HM/telomeric and rDNA silencing and that HST family members, including the widely expressed hSir2A, may perform evolutionarily conserved functions. PMID- 10473646 TI - The restriction site mutation assay: a review of the methodology development and the current status of the technique. AB - The restriction site mutation (RSM) assay has been employed in our laboratory, as a mutation detection system, since its first description in 1990. In principle the technique is capable of detecting mutations in ubiquitous restriction enzyme sites and is, therefore, readily applicable to any sequenced gene and/or organism. The RSM assay has been applied in our laboratory in various species, detecting rare mutations induced in mouse, rat, Xenopus, flatfish and human cells and tissues. This paper reviews the data accumulated by the RSM methodology in our hands and charts the developmental processes which have steadily improved the technique such that it is now applicable as a sensitive genotypic mutation detection system. This paper also includes PCR primer sequences and restriction enzymes employed in mutational analyses performed in the various species studied. We detail a variety of problems associated with the assay and the steps taken to solve them. The specific hurdles which have been overcome include the lack of quantitative data, the question of the contribution of DNA adducts to the induced mutation profile and the presence of false positives. Finally, the methods which have been developed to increase the sensitivity of the assay are also detailed. This paper describes our recommended RSM methodology, as it is routinely employed in our laboratory, which enables the analyses of mutations induced by chemical exposures and spontaneous endogenous processes. Our aim in presenting the developmental data on the RSM assay is to provide other researchers with sufficient information about the RSM methodology to facilitate its application in mutation analysis in other genes and organisms. PMID- 10473647 TI - Detection of mitomycin C-induced genetic damage in fish cells by use of RAPD. AB - Concern about genetic alterations in fish populations arising from anthropogenic activities has led to the adaptation and/or development of new tests and techniques that shed light on these alterations. The high number and the reduced size of chromosomes and the long cell cycle associated with most fish species preclude the use of most accepted genotoxicity assays. The purpose of this work was to study the capability of the randomly amplified polymorphic DNA technique to show genotoxic effects induced by chemicals in fish cells. To do that we studied the effect of 0.5 microg/ml mitomycin C (MMC) on an established rainbow trout cell line (RTG-2). To increase the sensitivity of detecting altered copies of DNA and to avoid the presence of false positives and a lack of reproducibility, the amounts of DNA template and primer present in amplification reactions were studied and optimized after comparison between the control and exposed fingerprints for 4, 6 and 8 h. Results show that 5 ng of DNA template and 4 pM chosen primer were optimum to show differences between control and exposed cells and to obtain reproducible results. The results obtained, after optimum conditions were established, show that this system could be useful for the assessment of DNA alterations in in vitro genotoxicity studies. PMID- 10473648 TI - Genotoxic activity of chlorohydroxyfuranones in the microscale micronucleus test on mouse lymphoma cells and the unscheduled DNA synthesis assay in rat hepatocytes. AB - Chlorohydroxyfuranones (CHFs) are mutagenic disinfection by-products found in chlorine-treated drinking water. In the current study, the genotoxicity of four CHFs, 3,4-dichloro-5-hydroxy-2(5H)-furanone (MCA), 3-chloro-4-methyl-5-hydroxy 2(5H)-furanone (MCF), 3-chloro-4-(chloromethyl)-5-hydroxy-2(5H)-furanone (CMCF) and 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), was determined. Two in vitro assays, the microscale micronucleus assay on L5178Y mouse lymphoma cells and the unscheduled DNA synthesis assay on a hepatocyte primary culture from Fisher F344 rats, were carried out. All four CHFs demonstrated genotoxic effects in both assays. In the two systems used, CMCF was the most genotoxic compound, followed by MCA, MX and MCF, respectively. This work was the first study of the DNA damaging properties of all four CHFs in two in vitro genotoxicity tests. These new data expand the range of genetic damages induced by this group of compounds. PMID- 10473649 TI - Genotoxicity testing of potassium canrenoate in cultured rat and human cells. AB - Potassium canrenoate (PC), a competitive aldosterone antagonist used as a diuretic and in the treatment of hypertension, was examined for its capacity to produce genotoxic effects in cultured rat and human cells. At subtoxic concentrations (10-90 microM) PC was found to induce a dose-dependent degree of DNA fragmentation, as detected by the Comet assay, and of DNA repair synthesis, as measured by quantitative autoradiography, in primary cultures of hepatocytes from rat and human donors of both genders. In rat hepatocytes both DNA fragmentation and DNA repair were more marked after 3 h than after 20 h exposure and in cultures from females than from males. In human hepatocytes from one male and two female donors, PC caused a similar effect in terms of DNA fragmentation, whereas DNA repair was detected in cultures from only two of the same three donors and was less marked than in rat hepatocytes. A modest but statistically significant increase in micronucleated cells was present in primary cultures of replicating rat hepatocytes exposed to 10 or 30 microM PC for 48 h, the response being, in this case also, more evident in females than in males. In contrast, PC did not induce micronucleus formation in human hepatocytes from two female donors. Any evidence of DNA fragmentation and micronucleus formation was absent in cultured human lymphocytes. Taken as a whole these findings support the hypothesis that hepatocytes activate PC to DNA-damaging reactive species. PC induced the observed genotoxic effects at concentrations close to those produced in humans by the administration of therapeutic doses, but these effects were as a whole more marked in rat than in human hepatocytes. Since PC shares the 17 hydroxy-3-oxopregna-4,6-diene structure with cyproterone acetate, chlormadinone acetate and megestrol acetate, previously found to be genotoxic to both rat and human hepatocytes, the potential carcinogenic hazard of this type of steroids cannot be neglected. PMID- 10473650 TI - Biodosimetry results obtained by various cytogenetic methods and electron spin resonance spectrometry among inhabitants of a radionuclide contaminated area around the siberian chemical plant (Tomsk-7). AB - On April 6, 1993, near the town of Tomsk (Russia) there was an accident at the Siberian Chemical Plant (SCP) which resulted in extensive contamination of an area of 250 km(2) to the north of SCP with long-lived radionuclides such as (239)Pu, (137)Cs and (90)Sr. Cytogenetic methods and electron spin resonance (ESR) spectrometry of tooth enamel were used to estimate the radiation doses received by the population. The ESR signal intensity and the chromosomal aberration frequency in lymphocytes of the tooth donors showed a good correlation. The data showed that 15% of the inhabitants of the Samus settlement received a radiation dose >90 cGy. The exceptions were results of an examination of fishermen, where ESR gave high values (80-210 cGy) but both the chromosome assay and the cytokinesis block micronucleus method gave lower ones (8-52 cGy). A large increase in chromosome damage was observed in people born between 1961 and 1969. It was found that during these years several serious accidents at the Siberian Chemical Plant had occurred causing radiation pollution of the area. The number of cells with chromosome aberrations was significantly less among the people arriving in Samus after 1980. We found good correlations between the level of carotene consumption and a decrease in frequency of both micronuclei in binucleated lymphocytes (r = 0.68, P < 0.01) and chromatid aberrations (r = 0.61, P < 0.01) among the inhabitants. We also examined the inhabitants of Samus for opisthorchis infection, which was present in 30% of the population. The Samus inhabitants affected by Opisthorchis felineus showed significantly increased levels of micronuclei in binucleated lymphocytes and chromatid aberrations as compared with the controls. PMID- 10473652 TI - Previous loss of chromosome 11 containing a suppressor locus increases radiosensitivity, neoplastic transformation frequency and delayed death in HeLa x fibroblast human hybrid cells. AB - CGL1 (HeLa x fibroblast) hybrid cells have been utilized to study mechanisms of radiation-induced neoplastic transformation of human cells in vitro. Previous analysis has shown that loss of active tumor suppressor alleles on fibroblast chromosomes 11 and 14 may be required for radiation-induced neoplastic transformation of CGL1 cells. Loss of chromosome 11 alone was, therefore, found to be necessary but not sufficient for neoplastic transformation. We postulated that the loss of chromosome 11 may make the hybrid cells more susceptible to radiation-induced neoplastic transformation, since these cells have already undergone one of the required tumor suppressor loss events. Hybrid cells which have lost one copy of chromosome 11 were designated CON104(-11). CON104(-11) hybrid cells were found to have increased X-ray sensitivity and susceptibility to radiation-induced neoplastic transformation when compared with the parental CGL1 cells. In addition, the neoplastically transformed foci appear to arise earlier after radiation exposure in CON104(-11) versus CGL1 cells. Furthermore, the plating efficiency (PE) of the progeny of the irradiated CON104(-11) cells, growing in transformation flasks, is persistently lower than parental CGL1 cells during the 21 day assay period. The lower PE of the progeny of irradiated cells was attributed to the expression of delayed death/lethal mutations post irradiation, a reflection of genomic instability. Taken together, the data indicate that previous loss of chromosome 11 may increase the radiation-induced genomic instability of the hybrid cells, leading to increased radiation sensitivity and neoplastic transformation potential. The data suggest that one possible function of the chromosome 11 tumor suppressor gene may be to help maintain genome stability after radiation damage. PMID- 10473651 TI - Induction of genetic damage in human lymphocytes and mutations in Salmonella by trihalomethanes: role of red blood cells and GSTT1-1 polymorphism. AB - The brominated trihalomethanes (THMs) are mutagenic and carcinogenic disinfection by-products frequently found in chlorinated drinking water. They can be activated to mutagens by the product of the glutathione S-transferase-Theta (GSTT1++-1) gene in Salmonella RSJ100, which has been transfected with this gene. To evaluate this phenomenon in humans, we have examined the genotoxicity of a brominated THM, bromoform (BF), using the Comet assay in human whole blood cultures exposed in vitro. No differences were found in the comet tail length between cultures from GSTT1-1(+) versus GSTT1-1(-) individuals (1.67 +/- 0.40 and 0.74 +/- 0.54 microm/mM, respectively, P = 0.28). The high variability was due to the relatively weak induction of comets by BF. Combining the data from both genotypic groups, the genotoxic potency of BF was 1.20 +/- 0.34 microm/mM (P = 0.003). GSTT1-1 is expressed in red blood cells but not in the target cells (lymphocytes), and expression within the target cell (as in Salmonella RSJ100) may be necessary for enhanced mutagenesis in GSTT1-1(+) relative to GSTT1-1(-) cultures. To examine this, we exposed Salmonella RSJ100 and a control strain not expressing the gene (TPT100) to the most mutagenic brominated THM detected in Salmonella, dibromochloromethane (DBCM), either in the presence or absence of S9 or red blood cells from GSTT1-1(+) or GSTT1-1(-) individuals. S9 did not activate DBCM in the non-expressing strain TPT100, and it did not affect the ability of the expressing strain RSJ100 to activate DBCM. As with S9, red cells from either genotypic group were unable to activate DBCM in TPT100. However, red cells (whole or lysed) from both genotypic groups completely repressed the ability of the expressing strain RSJ100 to activate DBCM to a mutagen. Such results suggest a model in which exposure to brominated THMs may pose an excess genotoxic risk in GSTT1-1(+) individuals to those organs and tissues that both express this gene and come into direct contact with the brominated THM, such as the colon. In contrast, those organs to which brominated THMs would be transported via the blood might be protected by erythrocytes. Such a proposal is reasonably consistent with the organ specificity of drinking water-associated cancer in humans, which shows slightly elevated risks for cancer of the colon and bladder but not of the liver. PMID- 10473653 TI - Analysis of chromosome loss and non-disjunction in cytokinesis-blocked lymphocytes of 24 male subjects. AB - Chromosome malsegregation in peripheral blood lymphocytes of 24 healthy male subjects was analysed by means of fluorescence in situ hybridization with centromeric probes of chromosomes 7, 11, 18 and X. On the basis of the distribution of centromeric signals in cytokinesis-blocked cells, both loss (leading to centromere-positive micronuclei) and non-disjunction (resulting in an unbalanced distribution of signals in the main nuclei) of the hybridized chromosomes in vitro were identified. In addition, the incidence of binucleated cells with two hyperploid nuclei, possibly arising from mitotic division of trisomic types, was determined. In this way, the incidence of chromosome malsegregation in vivo and in vitro could be compared in the same cell samples. The results obtained show that ageing is positively correlated with the incidence of malsegregation of chromosome X in peripheral lymphocytes of male subjects and confirm the higher susceptibility of chromosome X to malsegregation in comparison with autosomes. A positive correlation between in vitro and in vivo malsegregation rates was observed for both chromosome X and for autosomes. Finally, relatively high frequencies of multiple malsegregation events, greater than expected for independent events, were recorded for both chromosome X and for autosomes, indicating that the abnormal segregation of chromosomes may be connected to a general dysfunction of the mitotic apparatus. The correlation observed between in vitro and in vivo malsegregation frequencies and the association of both parameters with ageing suggest that analysis of chromosome malsegregation in binucleated cells is a useful tool in the study of genomic instability in human populations. PMID- 10473655 TI - The effect of ascorbate and alpha-tocopherol supplementation in vitro on DNA integrity and hydrogen peroxide-induced DNA damage in human spermatozoa. AB - The aim of this study was to determine the effects of supplementation with ascorbate and alpha-tocopherol, both singly and in combination, during sperm preparation on subsequent sperm DNA integrity, induced DNA damage and reactive oxygen species (ROS) generation. Semen samples with normozoospermic and asthenozoospermic profiles (n = 15 for each control and antioxidant group) were prepared by Percoll density centrifugation (95.0-47.5%) where the medium had been supplemented with these antioxidants to a number of different concentrations, all within physiological levels. Controls were included which had no ascorbate or alpha-tocopherol added. DNA damage was induced using hydrogen peroxide (H(2)O(2)) and DNA integrity was determined using a modified alkaline single cell gel electrophoresis (Comet) assay, while ROS generation was measured using chemiluminescence. Addition of ascorbate to sperm preparation medium did not affect baseline DNA integrity but did provide sperm with complete protection against H(2)O(2)-induced DNA damage. Generation of H(2)O(2)-induced ROS was also significantly reduced after treatment with ascorbate, although baseline levels were unaffected by this antioxidant. Supplementation of sperm preparation medium with alpha-tocopherol did not influence baseline DNA integrity but provided sperm with dose-dependent protection against H(2)O(2)-induced DNA damage. Generation of H(2)O(2)-induced ROS was significantly reduced after treatment with alpha tocopherol, although baseline ROS levels were unaffected by this antioxidant. Addition of both ascorbate and alpha-tocopherol in combination to sperm preparation medium actually induced DNA damage and intensified the damage induced by H(2)O(2), however, H(2)O(2)-induced ROS production was significantly reduced in a dose-dependent manner by supplementation with both vitamins. PMID- 10473656 TI - Induction of polyploidy and apoptosis after exposure to high concentrations of the spindle poison nocodazole. AB - The proportions of aneuploid/polyploid versus euploid cells formed after treatment with spindle poisons like nocodazole are of course dependent on the relative survival of cells with numerical chromosome aberrations. This work aimed at studying the survival of polyploid cells formed after treatment with a nocodazole concentration sufficient to significantly decrease tubulin polymerization (0.1 microg/ml). First, normal primary lymphocytes were analysed and the following complementary chromosomal parameters were quantified: mitotic index, frequency of abnormal mitoses, polyploid metaphases and apoptotic cells. The results clearly indicate a positive correlation between abnormal mitotic figures, apoptosis and the induction of polyploidy. They therefore led to a single cell approach in which both apoptosis and polyploidy induction could be scored in the same cell. For this purpose, actively proliferating cells are required and two human leukaemic cell lines were used, KS (p53-positive) and K562 (p53-negative), which have a near-triploid karyotype. Cells were separated into an apoptotic and a viable fraction by means of annexin-V staining and flow cytometry. In KS, treatment with nocodazole induced a similar fraction of hexaploid cells in both the viable and apoptotic fraction, but no dodecaploid cells were ever observed. In contrast, a population of dodecaploid cells (essentially viable) was clearly observed in the K562 cell line. The results in KS, as compared with K562, confirm that wild-type p53 can prevent further cycling of polyploid cells by blocking rereplication. The most probable explanation for these data is that not only the mitotic spindle but also interphase microtubules are sensitive to nocodazole treatment. Our data thus strongly suggest that besides the G(1)/S checkpoint under the control of p53, the G(2)/M transition may be sensitive to depolymerization of microtubules, possibly under the control of Cdc2, Bcl-2, Raf-1 and/or Rho. PMID- 10473654 TI - Processing of DNA damage induced by hydrogen peroxide and methyl methanesulfonate in human lymphocytes: analysis by alkaline single cell gel electrophoresis and cytogenetic methods. AB - The persistence of induced DNA damage in human lymphocytes after mitogen stimulation and its relationship to subsequent cytogenetic alterations were investigated. The analysis of single-strand breaks and alkali-labile sites by single cell gel electrophoresis (SCGE) showed the almost complete repair of damage induced in resting lymphocytes by methyl methanesulfonate (MMS, 140-210 microM) and hydrogen peroxide (H(2)O(2), 25-100 microM) during the first 16 h of culture. On the other hand, DNA damage was shown to persist to a large extent when cells were cultured in the presence of the repair inhibitor cytosine beta-D arabinofuranoside (Ara-C) (1 microg/ml). Although highly effective in the induction of DNA lesions detectable by SCGE, both agents failed to significantly increase the rate of micronucleus formation in cytokinesis-blocked cells harvested 66 h after treatment. However, when Ara-C was present during the first 16 h of culture, micronuclei were significantly increased at all doses. Conversely, sister chromatid exchange (SCE) rates were increased by chemical treatments to a higher extent in cultures without Ara-C. Delayed treatments, 16 h after mitogen stimulation, led to a significant induction of micronuclei in the case of MMS but not with H2O(2). These results suggest that only a minor fraction of DNA damage induced in resting lymphocytes is available for fixation through misreplication, because of its effective repair prior to S phase. However, the processing of damage through recombination pathways can lead to increased SCE rates in treated cells. These features of the processing of DNA damage in human lymphocytes should be taken into account when structural cytogenetic alterations in cultured lymphocytes are used in monitoring human exposure to genotoxic agents. PMID- 10473658 TI - Sports injuries: emergency assessment and field-side care. PMID- 10473657 TI - Meeting report. Joint COC/COM symposium on genetic susceptibility to cancer, Department of Health, London, UK, October 1998. PMID- 10473659 TI - Pediatric septic shock. PMID- 10473660 TI - Index of suspicion. Case 1. Alagille syndrome. PMID- 10473661 TI - Earning CME Credit-Completing the PIR Quiz. PMID- 10473662 TI - The new biology enters the generalist pediatrician's office: lessons from the Human Genome Project. PMID- 10473663 TI - Consultation with the specialist: Eye trauma: corneal abrasions. PMID- 10473664 TI - Hepatitis C. PMID- 10473665 TI - The importance of early nutritional management of low-birthweight infants. PMID- 10473666 TI - Early aggressive nutrition in the neonate. PMID- 10473667 TI - Nutrition for ill neonates. PMID- 10473668 TI - c-Jun triggers apoptosis in human vascular endothelial cells. AB - In endothelial cells (ECs), the transcription factor c-Jun is induced by a variety of stimuli that perturb EC function. To extend our understanding of the role of c-Jun in EC physiology, we have directed overexpression of c-Jun in human umbilical vein ECs by using a tetracycline-regulated adenoviral expression system. In this study, we report a novel observation using this system. Specific expression of c-Jun is a sufficient trigger for ECs to undergo apoptosis, as demonstrated by a set of combined assays including an ELISA specific for histone associated DNA fragmentation, DNA laddering, and TdT-mediated dUTP nick end labeling (TUNEL). Tetracycline can effectively shut off c-Jun overexpression and prevent EC apoptosis. Cleavage of poly(ADP-ribose) polymerase was also detected in ECs overexpressing c-Jun. Moreover, inhibitors of cysteine proteases blocked the apoptosis, suggesting a caspase-associated mechanism involved in proapoptotic effects of c-Jun. To gain further insight into the role of c-Jun as a pathophysiological regulator of EC death, TAM67, a dominant-negative mutant of c Jun, was overexpressed in human umbilical vein ECs to abrogate endogenous c Jun/activator protein-1 activation. H(2)O(2)-triggered apoptosis was largely attenuated in ECs overexpressing TAM67. Together, these results suggest that c Jun, as a proapoptotic molecule, may play a role in mediating the cell death program in vascular endothelium. PMID- 10473669 TI - Peroxisome proliferator-activated receptor activators inhibit thrombin-induced endothelin-1 production in human vascular endothelial cells by inhibiting the activator protein-1 signaling pathway. AB - Endothelin-1 (ET-1), a 21-amino acid vasoactive peptide mainly produced by vascular endothelial cells, is involved in the regulation of vascular tone and smooth muscle cell proliferation. Peroxisome proliferator-activated receptors (PPARs), key players in lipid and glucose metabolism, have been implicated in metabolic disorders that are predisposing to atherosclerosis. Because of the potential role of ET-1 in vascular disorders such as hypertension and atherosclerosis, we investigated the regulation of ET-1 expression by PPAR activators. Western blot and reverse transcription-polymerase chain reaction analyses demonstrated that both PPARalpha and PPARgamma are expressed in human coronary artery endothelial cells as well as in endothelial cell lines such as HMEC-1 and ECV304. In bovine aortic endothelial cells and HMEC-1 cells, both PPARalpha and PPARgamma ligands inhibited thrombin-induced ET-1 secretion, whereas basal ET-1 secretion was only slightly suppressed. Reverse transcription polymerase chain reaction experiments showed that this inhibition of ET-1 production occurs at the gene expression level. Using transient transfection assays, we demonstrated that PPARs downregulate thrombin-activated transcription of the human ET-1 promoter. Transactivation studies with c-Jun and c-Fos expression plasmids indicated that PPARs negatively interfere with the activator protein-1 signaling pathway, which mediates thrombin activation of ET-1 gene transcription. Furthermore, electrophoretic mobility shift assays demonstrated that PPAR activators reduce the thrombin-stimulated binding activity of bovine aortic endothelial cell nuclear extracts as well as c-Jun binding to an activator protein-1 consensus site. Taken together, these data indicate that (1) both PPARalpha and PPARgamma are expressed in human vascular endothelial cells and (2) PPAR activators inhibit thrombin-induced ET-1 biosynthesis, indicating a novel role for PPARs in vascular endothelial function. PMID- 10473670 TI - The mitochondrial apoptotic pathway is activated by serum and glucose deprivation in cardiac myocytes. AB - Many cell types undergo apoptosis under conditions of ischemia. Little is known, however, about the molecular pathways that mediate this response. A cellular and biochemical approach to elucidate such signaling pathways was undertaken in primary cultures of cardiac myocytes, a cell type that is especially sensitive to ischemia-induced apoptosis. Deprivation of serum and glucose, components of ischemia in vivo, resulted in myocyte apoptosis, as determined by nuclear fragmentation, internucleosomal cleavage of DNA, and processing of caspase substrates. These manifestations of apoptosis were blocked by zVAD-fmk, a peptide caspase inhibitor, indicating that caspase activity is necessary for the progression of apoptosis in this model. In contrast to control cells, apoptotic myocytes exhibited cytoplasmic accumulation of cytochrome c, indicating release from the mitochondria. Furthermore, both caspase-9 and caspase-3 were processed to their active forms in serum-/glucose-deprived myocytes. Caspase processing, but not cytochrome c release, was inhibited by zVAD-fmk, placing the latter event upstream of caspase activation. This evidence demonstrates that components of ischemia activate the mitochondrial death pathway in cardiac myocytes. PMID- 10473671 TI - Subcellular [Ca2+]i gradients during excitation-contraction coupling in newborn rabbit ventricular myocytes. AB - The central role of T-tubule and sarcoplasmic reticulum (SR) diadic junctions in excitation-contraction (EC) coupling in adult (AD) ventricular myocytes suggests that their absence in newborn (NB) cells may manifest as an altered EC coupling phenotype. We used confocal microscopy to compare fluo-3 [Ca2+]i transients in the subsarcolemmal space and cell center of field-stimulated NB and AD rabbit ventricular myocytes. Peak systolic [Ca2+]i occurred sooner and was higher in the subsarcolemmal space compared with the cell center in NB myocytes. In AD myocytes, [Ca2+]i rose and declined with similar profiles at the cell center and subsarcolemmal space. Disabling the SR (10 micromol/L thapsigargin) slowed the rate of rise and decline of Ca2+ in AD myocytes but did not alter Ca2+ transient kinetics in NB myocytes. In contrast to adults, localized SR Ca2+ release events ("Ca2+ sparks") occurred predominantly at the cell periphery of NB myocytes. Immunolabeling experiments demonstrated overlapping distributions of the Na(+) Ca2+ exchanger and ryanodine receptors (RyR2) in AD myocytes. In contrast, RyR2s were spatially separated from the sarcolemma in NB myocytes. Confocal sarcolemmal imaging of di-8-ANEPPS-treated myocytes confirmed an extensive T-tubule network in AD cells, and that T-tubules are absent in NB myocytes. A mathematical model of subcellular Ca2+ dynamics predicts that Ca2+ flux via the Na(+)-Ca2+ exchanger during an action potential can account for the subsarcolemmal Ca2+ gradients in NB myocytes. Spatial separation of sarcolemmal Ca2+ entry from SR Ca2+ release channels may minimize the role of SR Ca2+ release during normal EC coupling in NB ventricular myocytes. PMID- 10473673 TI - Intravenous allopurinol decreases myocardial oxygen consumption and increases mechanical efficiency in dogs with pacing-induced heart failure. AB - Allopurinol, an inhibitor of xanthine oxidase, increases myofilament calcium responsiveness and blunts calcium cycling in isolated cardiac muscle. We sought to extend these observations to conscious dogs with and without pacing-induced heart failure and tested the prediction that allopurinol would have a positive inotropic effect without increasing energy expenditure, thereby increasing mechanical efficiency. In control dogs (n=10), allopurinol (200 mg IV) caused a small positive inotropic effect; (dP/dt)(max) increased from 3103+/-162 to 3373+/ 225 mm Hg/s (+8.3+/-3.2%; P=0.01), but preload-recruitable stroke work and ventricular elastance did not change. In heart failure (n=5), this effect was larger; (dP/dt)(max) rose from 1602+/-190 to 1988+/-251 mm Hg/s (+24.4+/-8.7%; P=0.03), preload-recruitable stroke work increased from 55.8+/-9.1 to 84. 9+/ 12.2 mm Hg (+28.1+/-5.3%; P=0.02), and ventricular elastance rose from 6.0+/-1.6 to 10.5+/-2.2 mm Hg/mm (P=0.03). Allopurinol did not affect myocardial lusitropic properties either in control or heart failure dogs. In heart failure dogs, but not controls, allopurinol decreased myocardial oxygen consumption (-49+/-4.6%; P=0. 002) and substantially increased mechanical efficiency (stroke work/myocardial oxygen consumption; +122+/-42%; P=0.04). Moreover, xanthine oxidase activity was approximately 4-fold increased in failing versus control dog hearts (387+/-125 versus 78+/-72 pmol/min. mg(-1); P=0.04) but was not detectable in plasma. These data indicate that allopurinol possesses unique inotropic properties, increasing myocardial contractility while simultaneously reducing cardiac energy requirements. The resultant boost in myocardial contractile efficiency may prove beneficial in the treatment of congestive heart failure. PMID- 10473672 TI - Atrial L-type Ca2+ currents and human atrial fibrillation. AB - Chronic atrial fibrillation (AF) is characterized by decreased atrial contractility, shortened action potential duration, and decreased accommodation of action potential duration to changes in activation rate. Studies on experimental animal models of AF implicate a reduction in L-type Ca2+ current (I(Ca)) density in these changes. To evaluate the effect of AF on human I(Ca), we compared I(Ca) in atrial myocytes isolated from 42 patients in normal sinus rhythm at the time of cardiac surgery with that of 11 chronic AF patients. I(Ca) was significantly reduced in the myocytes of patients with chronic AF (mean 3.35+/-0.5 pA/pF versus -9.13+/-1. 0 pA/pF in the controls), with no difference between groups in the voltage dependence of activation or steady-state inactivation. Although I(Ca) was lower in myocytes from the chronic AF patients, their response to maximal beta-adrenergic stimulation was not impaired. Postoperative AF frequently follows cardiac surgery. Half of the patients in the control group (19/38) of this study experienced postoperative AF. Whereas chronic AF is characterized by reduced atrial I(Ca), the patients with the greatest I(Ca) had an increased incidence of postoperative AF, independent of patient age or diagnosis. This observation is consistent with the concept that calcium overload may be an important factor in the initiation of AF. The reduction in functional I(Ca) density in myocytes from the atria of chronic AF patients may thus be an adaptive response to the arrhythmia-induced calcium overload. PMID- 10473674 TI - Absence of troponin I degradation or altered sarcoplasmic reticulum uptake protein expression after reversible ischemia in swine. AB - The findings of troponin I (TnI) proteolysis (in isolated rat hearts) and induction of selected sarcoplasmic reticulum (SR) calcium-regulatory genes (after repetitive total coronary occlusions in swine) have given rise to the hypothesis that the time course of functional recovery of stunned myocardium reflects the resynthesis of reversibly damaged proteins. Although stunning occurs after brief total occlusions and prolonged partial occlusions (ie, short-term hibernation), the time course of functional recovery varies from a few hours to several days, suggesting that the severity of protein damage or mechanisms responsible for the dysfunction may differ. To study this, we examined SR gene expression and TnI degradation in stunned myocardium produced by 10-minute total left anterior descending coronary artery (LAD) occlusions (n=4) or 1-hour partial LAD occlusions, in which flow was reduced to approximately 50% of control values for 60 minutes (n=6) in swine. One hour after reperfusion, LAD wall thickening was severely depressed in both models despite normal perfusion and no triphenyltetrazolium chloride evidence of necrosis. Normal myocardium exhibited TnI immunoreactivity at 31 kDa and a weak secondary band at 27 kDa. Irreversible injury or calpain activation in vitro produced a marked increase in the intensity of the 27-kDa band, consistent with TnI degradation. Stunned myocardium demonstrated no change in the 31- or the 27-kDa band, and the percentage of the 27- to 31-kDa band remained constant after 10-minute total occlusions (LAD, 5.9+/ 0.9%; normal, 4.9+/-1.6%) and 1-hour partial occlusions (LAD, 8.5+/-1.9%; normal, 7.3+/-1.4%) and in sham controls (LAD, 10.9+/-1.5%; normal, 9.8+/-1.4%). Northern analysis showed no alterations in TnI or SR gene expression, but the stress protein HSP-70 was variably induced. Thus, stunned myocardium occurs without TnI degradation or altered SR gene expression, indicating that additional mechanisms are responsible for the reversible dysfunction after single episodes of regional ischemia in swine. PMID- 10473675 TI - Role of respiratory motor output in within-breath modulation of muscle sympathetic nerve activity in humans. AB - We measured muscle sympathetic nerve activity (MSNA, peroneal microneurography) in 5 healthy humans under conditions of matched tidal volume, breathing frequency, and end-tidal CO(2), but varying respiratory motor output as follows: (1) passive positive pressure mechanical ventilation, (2) voluntary hyperventilation, (3) assisted mechanical ventilation that required the subject to generate -2.5 cm H(2)O to trigger each positive pressure breath, and (4) added inspiratory resistance. Spectral analyses showed marked respiratory periodicities in MSNA; however, the amplitude of the peak power was not changed with changing inspiratory effort. Time domain analyses showed that maximum MSNA always occurred at end expiration (25% to 30% of total activity) and minimum activity at end inspiration (2% to 3% of total activity), and the amplitude of the variation was not different among conditions despite marked changes in respiratory motor output. Furthermore, qualitative changes in intrathoracic pressure were without influence on the respiratory modulation of MSNA. In all conditions, within-breath changes in MSNA were inversely related to small changes in diastolic pressure (1 to 3 mm Hg), suggesting that respiratory rhythmicity in MSNA was secondary to loading/unloading of carotid sinus baroreceptors. Furthermore, at any given diastolic pressure, within-breath MSNA varied inversely with lung volume, demonstrating an additional influence of lung inflation feedback on sympathetic discharge. Our data provide evidence against a significant effect of respiratory motor output on the within-breath modulation of MSNA and suggest that feedback from baroreceptors and pulmonary stretch receptors are the dominant determinants of the respiratory modulation of MSNA in the intact human. PMID- 10473677 TI - Ionic determinants of atrial fibrillation and Ca2+ channel abnormalities : cause, consequence, or innocent bystander? PMID- 10473676 TI - In search of the proteins that cause myocardial stunning. PMID- 10473678 TI - Logarithmic 3-Band Color Encoding: Robust Method for Display and Comparison of Compositional Maps in Electron Probe X-ray Microanalysis. AB - : Electron-excited X-ray maps recorded with the scanning electron microscope (SEM)/electron probe X-ray microanalyzer (EPMA) are a major method of presenting compositional information. Digitally recorded maps are processed in a variety of ways to improve the visibility of features. Scaling of the recorded signal to match the 8-bit gray-scale intensity range of a typical computer display system is almost always necessary. Inherent limitations of gray-scale displays have led to other intensity-encoding methods for X-ray maps, including clipping, histogram normalization, and pseudocolor scales. While feature visibility is improved by applying these scales, comparisons among image sets are difficult. Quantitative comparisons must be based on standardized intensities corrected for background to produce intensity ratio (k-value) maps. We have developed a new logarithmic, multiband color-encoding method to view these k-value maps more effectively. Three color bands are defined, starting with a dark primary color and grading to a bright pastel: blue = trace (0.001 to 0.01); green = minor (0.01 to 0.1); and red = major (0.1 to 1.0). Within each band, the color is assigned according to a logarithmic scale that depends on intensity ratio or compositional measurements. Logarithmic multiband color encoding permits direct comparisons of maps, such as maps of different elements in the same field of view or maps of the same element in different areas, because the color scale is identical for all maps. PMID- 10473679 TI - Cytokine and Eicosanoid Production by Cultured Human Monocytes Exposed to Titanium Particulate Debris. AB - : Phagocytosis of particulate wear debris from arthroplasties by macrophages induces an inflammatory response that has been linked to implant loosening and premature failure of artificial joints. Inflammatory mediators released by phagocytic macrophages such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and prostaglandin E(2) (PGE(2)) are believed to play a central role in the pathogenesis of aseptic loosening. The objective of this study was to characterize titanium alloy particulates that closely match wear debris found around joint arthroplasties and to study their effects on the biosynthesis of inflammatory mediators by cultured monocytes. Peripheral blood monocytes were isolated from healthy human volunteers. Monocytes were cultured in 96-well plates for 24 h, washed, and exposed to three concentrations of titanium particulates and controls from 18-24 h. Supernatants were assayed for TNF-alpha, IL-1beta, IL-6, and PGE(2) activity. Energy dispersive X-ray spectroscopy (EDX) verified the titanium alloy to be Ti6A14V. Scanning electron microscopy (SEM) analysis showed significant titanium particulate heterogeneity with approximately 95% of the particles <1 um in diameter. SEM and EDX technology was useful in the characterization of the titanium particulates utilized for in vitro models of titanium-induced cytokine release by monocytes. Incubation of titanium particulates (in concentrations similar to those found around loosened prosthetic joints) with cultured monocytes significantly increased their production of TNF-alpha, IL-1beta, and PGE(2). PMID- 10473680 TI - Atomic Structures of Oxygen-associated Defects in Sintered Aluminum Nitride Ceramics. AB - : Convergent-beam electron diffraction and Z-contrast imaging are used to study oxygen-associated defects, flat inversion domain boundaries, dislocations, and interfaces in sintered AlN ceramics. The structures of these defects are directly derived from atomic-resolution Z-contrast images. The flat inversion domain boundaries contain a single Al-O octahedral layer and have a stacking sequence of.bAaB-bAc-CaAc., where -cAb- indicates the single octahedral layer. The expansion at the flat inversion domain boundaries is measured to be 0.06 (+/ 0.02) nm. The interfaces between 2H- and polytypoid-AlN are found to be also inversion domain boundaries but their stacking sequence differs from that of the flat inversion domain boundaries. PMID- 10473681 TI - Low-loss Electron Energy-loss Spectroscopy and Dielectric Function of Biological and Geological Polymorphs of CaCO(3). AB - : Previous work on microstructural characterization has shown variations in terms of defects and organization of nanostructures in the two polymorphs of calcium carbonate, calcite, and aragonite in mollusc shells. Large variations in mechanical properties are observed between these sections which have been attributed to variations in composite microstructure as well as intrinsic properties of the inorganic phases. Here we present local low-loss electron energy-loss spectroscopic (EELS) study of calcitic and aragonitic regions of abalone shell that were compared to geological (single-crystal) counterpart polymorphs to reveal intrinsic differences that could be related to organismal effects in biomineralization. In both sets of samples, local dielectric function is computed using Kramer-Kronig analysis. The electronic structures of biogenic and geological calcitic materials are not significantly different. On the other hand, electronic structure of biogenic aragonite is remarkably different from that of geological aragonite. This difference is attributed to the increased contribution from single electron excitations in biogenic aragonite as compared to that of geological aragonite. Furthermore, an apparent bound characteristic of the Re(1/epsilon) is observed for biological samples which suggests a "quasi plasmon"-like nature of the collective excitations. Implications of these changes are discussed in the context of macromolecular involvement in the making of the microstructures and properties in biogenic phases. PMID- 10473683 TI - Microsc. Microanal., Vol. 5, No. 3, May/June 1999, pp. 173-186. PMID- 10473684 TI - News and Commentary. PMID- 10473682 TI - Method for Cross-sectional Transmission Electron Microscopy Specimen Preparation of Composite Materials Using a Dedicated Focused Ion Beam System. AB - : A new method for transmission electron microscope (TEM) specimen preparation using a focused ion beam (FIB) system that results in a lower rate of gallium (Ga) implantation has been developed. The method was applied to structural and analytical studies of composite materials such as silicon (Si)-devices and magneto-optical disk. To protect the specimens against Ga ion irradiation, amorphous tungsten (W) was deposited on the surface of the specimen prior to FIB milling. The deposition was quite effective in reducing the Ga implantation rate, and energy-dispersive X-ray (EDX) analysis of these specimens detected 0.3-1.5% Ga incorporated in the thinned area. FIB milling times for these specimens were 1.5-2 hr, with a starting thickness of about 50 um. Although the milling rate was high, all the materials were properly prepared for TEM study, and clear crystal lattice images were observed on all specimens. PMID- 10473714 TI - COIN guidelines PMID- 10473716 TI - Tumour markers: same marker, different assay, different result. PMID- 10473715 TI - Radionuclide therapy. PMID- 10473717 TI - The MRC's commitment to cancer trials. PMID- 10473718 TI - Patient information materials in oncology: are they needed and do they work? PMID- 10473719 TI - Survey of attitudes of UK clinical oncologists towards radionuclide therapy. AB - A survey of UK consultants in clinical oncology was carried out in November 1997. The aim of this was to explore their attitudes towards radionuclide therapy and to determine the proportion of clinical oncologists involved in this modality. Three hundred and twenty-eight questionnaires were sent out and 227 (69.2%) were returned. Approximately one-half of those responding treat thyroid cancer or benign thyroid disease with radioactive iodine ((131)I) or prostate cancer with strontium ((89)Sr). The median number of patients treated per year with (131)I for benign thyroid disease was 12, with evidence of increasing subspecialization. Treatment with radioactive phosphorus ((32)P) for haematological disorders and yttrium ((90)Y) for intra-articular conditions was carried out by 31% (median number treated per year, two) and 6% (median number per year, five) respectively. There was strong support for the continuing involvement of clinical oncologists in radionuclide therapy for thyroid and prostate cancer. There was significant support for patients with benign thyroid disease or requiring intra-articular (90)Y to be treated by a specialist in nuclear medicine. However, this support was less strong amongst those currently involved in treatment compared with those not involved. There was support for the continued involvement of clinical oncologists in the use of (32)P. Attitudes appeared not to vary in different parts of the UK. PMID- 10473721 TI - Thyroid cancer in Wales 1985-1996: a cancer registry-based study. AB - No dedicated clinicopathological database for thyroid cancer exists in Wales to provide information about the incidence, histology, treatment and outcome of this uncommon malignancy. We have used cancer registry data to examine the epidemiology of thyroid cancer in Wales between 1985 and 1996 and have searched the database for clinically relevant data about stage, treatment and survival. A total of 699 cases were identified in this 12-year period, with no significant differences in their distribution between the five health authorities. Further analysis revealed a very wide age range (8-93 years) with a predominance of females (F:M ratio 2.8:1) and survival strongly influenced by gender, age and histological type. There were insufficient data to stage these cancers and only limited data about the surgical procedures undertaken. There was no information on non-surgical treatments (radio-iodine, radiotherapy and chemotherapy) in the database. Cancer registry data is well able to sustain an analysis of the epidemiology of thyroid cancer but further work is necessary to improve the quality of clinically relevant information about stage and treatment that could be used for audit. PMID- 10473720 TI - Thyroglobulin antibodies in differentiated thyroid cancer. AB - A retrospective review of patients with differentiated thyroid cancer (DTC) who were seen between 1984 and 1996 at the Royal Marsden Hospital identified 40 patients with serum thyroglobulin antibodies (TgAb). These antibodies can interfere with the immunoradiometric assay for serum thyroglobulin (Tg) used at this hospital, with resulting underestimation of the Tg level. A review of the case notes was carried out to ascertain the clinical significance of TgAb. The median follow-up from diagnosis of DTC was 26 months (range 3-401). The median age at diagnosis of DTC was 50 years (range 13-83). Patients were grouped according to the TgAb titre (high titre: TgAb >1/100, n = 28; low titre TgAb <1/100, n = 12). Thirteen patients relapsed, 11 in the high titre group and two in the low titre group. Sites of recurrence were: neck (n = 9), lung (n = 5), bone (n = 4) and brain (n = 2). No patient in the high titre group showed an elevated Tg with recurrence. One patient in the low titre group showed a Tg response to recurrence. Overall, the Tg assay failed to detect 92% of recurrences. Eight patients in the high titre group developed TgAb, apparently in response to tumour progression. In a third patient in the low titre group, the TgAb also acted as a 'tumour marker'. Thus, overall TgAb acted as a tumour marker in nine of the 40 (22.5%) patients in whom it was detected, and in nine of the 470 (1.9%) patients on follow-up during this time period. The overall survival of the whole group was 69% at 10 years. For patients with papillary carcinoma (n = 34) overall survival was 78% at 10 years. Laboratories should report routinely the presence of TgAb, with a caution indicating the direction of possible error (which depends on the assay used). Clinicians should appreciate that Tg measurements are unreliable in the presence of TgAb and that the development of TgAb can indicate active tumour. PMID- 10473722 TI - Radiotherapy in poor risk patients with stage I cancer of the endometrium: results of not giving external beam radiotherapy. AB - Poor prognosis (poorly differentiated and/or deep myometrial invasion) Stage I endometrial cancer can have a relapse rate as high as 50%. Traditionally, most clinical oncologists treat these patients with external beam radiotherapy after surgery but there is no evidence to show that this improves survival. The retrospective study looks at the results of not giving external beam radiotherapy in 25 consecutive patients and compares the results with a group of 13 consecutive patients who did have such treatment. The two groups were comparable with regard to age, degree of differentiation and degree of invasion. Survival was comparable in the two groups. There is no evidence of any obvious decrease in survival from withholding external beam radiotherapy, but this was not a prospective randomized controlled trial. This study illustrates that it is essential that the Medical Research Council ASTEC trial should be supported because this will determine the true place of external beam radiotherapy in such patients. PMID- 10473724 TI - Pathology of renal cell carcinoma: recent developments. PMID- 10473723 TI - Primary radiotherapy for carcinoma of the endometrium using external beam radiotherapy and single line source brachytherapy. AB - A small proportion of patients with adenocarcinoma of the endometrium are inoperable by virtue of severe concurrent medical conditions, gross obesity or advanced stage disease. They can be treated with primary radiotherapy with either curative or palliative intent. We report 37 such patients treated mainly with a combination of external beam radiotherapy and intracavitary brachytherapy using a single line source technique. The 5-year disease-specific survival for nonsurgically staged patients was 68.4% for FIGO Stages I and II and 33.3% for Stages III and IV. The incidence of late morbidity was acceptably low. Using the Franco-Italian Glossary, there was 27.0% grade 1 but no grade 2-4 bladder toxicity. For the rectum the rates were 18.9% grade 1, 5.4% grade 2, 2.7% grade 3, and no grade 4 toxicity. Methods of optimizing the dose distribution of the brachytherapy by means of variation of treatment length, radioactive source positions, and prescription point according to tumour bulk and individual anatomy are discussed. The biologically equivalent doses (BED) for combined external beam radiotherapy and brachytherapy were calculated to be in the range of 78-107 Gy(3) or 57-75 Gy(10) at point 'A' and appear adequate for the control of Stage I cancers. PMID- 10473725 TI - Failure of TSH rise prior to radio-iodine therapy for thyroid cancer: implications for treatment. AB - We describe three patients with well-differentiated thyroid carcinoma in whom no rise in serum thyroid-stimulating hormone (TSH) was observed after the discontinuation of thyroid hormone. In one patient, TSH deficiency was due to panhypopituitarism secondary to the empty sella syndrome. This patient initially failed to respond to (131)I but was subsequently given purified porcine TSH prior to further (131)I therapy. This resulted in a significant fall in the thyroglobulin level. In two further patients, TSH levels were suppressed by functioning follicular thyroid cancer. There was an unexpectedly good (131)I uptake by metastases and they responded clinically. The failure of TSH levels to rise after thyroid hormone withdrawal should prompt investigation of the pituitary-thyroid axis. In patients with hypopituitarism, exogenous TSH is recommended, to increase the (131)I uptake. In contrast, when TSH is suppressed by functioning tumour, radio-iodine treatment may still be effective. PMID- 10473726 TI - Bilateral cerebellopontine metastases in a patient with an unknown primary. AB - We report the case history of a patient who presented with symptoms and signs suggestive of a left cerebellopontine tumour. He underwent an exploratory posterior fossa craniotomy, which revealed metastatic adenocarcinoma. Despite intensive investigation, a primary site was not located and the patient received cranial irradiation. The patient re-presented 2 years later with further symptoms from a metastasis in the right cerebellopontine angle. He refused further intervention. PMID- 10473727 TI - Infusional 5-fluorouracil can be a pain in the neck: A case for repositioning displaced Hickman lines. AB - Increasing numbers of patients receive infusional chemotherapy or total parenteral nutrition via Hickman or Grochong lines. Although the insertion of these indwelling catheters is generally performed under radiological guidance and their positions verified by chest radiography, it is still feasible for them to become displaced at a later date. This possibility should be excluded in patients who develop unusual symptoms during the course of their infusional therapy. We review the reported complications associated with Hickman lines, and present a case history demonstrating that interventional radiology has a valuable role in displaced line repositioning, after the exclusion of thrombosis and infection. PMID- 10473729 TI - Pancreatic cancer in a young adult after treatment for Hodgkin's disease. AB - The authors present a case of adenocarcinoma of the head of the pancreas, which arose 14 years after extended field irradiation for Stage IIA Hodgkin's disease. The patient was aged 37 years at the time of the pancreatic cancer, which was situated within the previously irradiated volume. PMID- 10473728 TI - Metastatic renal oncocytoma: A case report and review of the literature. AB - Oncocytic renal tumour is an uncommon tumour and considered by many to be benign. Metastases are extremely rare. A case of renal oncocytoma with extensive skeletal metastases is described and the relevant literature reviewed. PMID- 10473731 TI - Compensation scheme for radiation linked diseases. PMID- 10473730 TI - Long-term survival following non-Hodgkin's lymphoma arising in Wiskott-Aldrich syndrome. AB - Wiskott-Aldrich syndrome (WAS) is a rare X-linked recessive trait, characterized by thrombocytopenia, eczema, immunodeficiency and a high risk of malignancy, usually leukaemia or lymphoma. Until recently, most patients died before the age of 10 years. A patient with WAS who developed extranodal non-Hodgkin's lymphoma at the age of 16 years is reported. Despite thrombocytopenia at presentation, chemotherapy was well tolerated. There was disease progression after first line chemotherapy and radiotherapy, but the patient responded to second line chemotherapy with cisplatin, vincristine and etoposide. He remains disease free 9 years after completing treatment. PMID- 10473733 TI - Manual dexterity and the making of tools - an introduction from an evolutionary perspective. PMID- 10473735 TI - Reflex pathways from large muscle spindle afferents and recurrent axon collaterals to motoneurones of wrist and digit muscles: a comparison in cats, monkeys and humans. AB - Whereas a large body of data is available on the control of hand motoneurones from the brain, not much is known about the contribution of the spinal interneuronal apparatus to the differentiated movement repertoire of the hand. This review summarises recent data on the excitatory Ia pathways and on recurrent inhibition for cats, monkeys and humans. The basic principles of organisation have, in general, been preserved in the different species. Thus the motoneurones to cat and human long and short digit muscles seem not to possess a recurrent axon collateral system. With regard to the Ia pathways, specialised connectivity patterns have developed in the long digit extensor and wrist extensor muscles. They allow the former group to support the differentiated movement repertoire of the digits, and the latter group to be part of a general extensor or flexor synergism. Modifications between the species are present, however, with regard to the proximodistal connectivity across the elbow. Whereas they are regularly present in the cat, they are less developed in the monkey and absent in man, which frees the human hand from the elbow position. PMID- 10473734 TI - Neural control of dexterity: what has been achieved? AB - This chapter reviews progress made in our understanding of the neural control of dexterity. It stresses the increasing benefit derived by uniting the different disciplines concerned with the study of the hand. It highlights the study of natural movements and of the importance of tackling the function of the interface between the neural control system and the biomechanical apparatus of the hand and arm. It also highlights the distributed nature of the control system, its utilisation of complex spatio-temporal representations and its dependence on sensory input. It concludes by pointing out the lessons that have been learned from two fields of work: the development of motor skill and the comparative study of dexterity in different primate species PMID- 10473736 TI - Control of grasp stability during pronation and supination movements. AB - We analyzed the control of grasp stability during a major manipulative function of the human hand: rotation of a grasped object by pronation and supination movement. We investigated the regulation of grip forces used to stabilize an object held by a precision grip between the thumb and index finger when subjects rotated it around a horizontal axis. Because the center of mass was located distal to the grip axis joining the fingertips, destabilizing torque tangential to the grasp surfaces developed when the grip axis rotated relative to the field of gravity. The torque load was maximal when the grip axis was horizontal and minimal when it was vertical. An instrumented test object, with a mass distribution that resulted in substantial changes in torque load during the rotation task, measured forces and torques applied by the digits. The mass distribution of the object was unpredictably changed between trials. The grip force required to stabilize the object increased directly with increasing torque load. Importantly, the grip force used by the subjects also changed in proportion to the torque load such that subjects always employed adequate safety margins against rotational slips, i.e., some 20-40% of the grip force. Rather than driven by sensory feedback pertaining to the torque load, the changes in grip force were generated as an integral part of the motor commands that accounted for the rotation movement. Subjects changed the grip force in parallel with, or even slightly ahead of, the rotation movement, whereas grip force responses elicited by externally imposed torque load changes were markedly delayed. Moreover, blocking sensory information from the digits did not appreciably change the coordination between movement and grip force. We thus conclude that the grip force was controlled by feedforward rather than by feedback mechanisms. These feedforward mechanisms would thus predict the consequences of the rotation movement in terms of changes in fingertip loads when the orientation of the grip axis changed in the field of gravity. Changes in the object's center of mass between trials resulted in a parametric scaling of the motor commands prior to their execution. This finding suggests that the sensorimotor memories used in manipulation to adapt the motor output for the physical properties of environmental objects also encompass information related to an object's center of mass. This information was obtained by somatosensory cues when subjects initially grasped the object with the grip axis vertical, i. e., during minimum tangential torque load. PMID- 10473737 TI - The effects of muscimol inactivation of small regions of motor and somatosensory cortex on independent finger movements and force control in the precision grip. AB - This study investigated the effects of inactivating small regions of the primary somatosensory (SI) and motor (MI) cortex on the control of finger forces in a precision grip. A monkey was trained to grasp and lift a computer-controlled object between the thumb and index finger and to hold it stationary within a narrow position window for 2 s. The grip force applied perpendicular to the object surface, the lifting or load force applied tangentially in the vertical direction, and the vertical displacement were sampled at 100 Hz. Also, the ability of the monkey to extract small pieces of food from narrow wells of a Kluver board was analyzed from video-tape. Preliminary single-unit recordings and microstimulation studies were used to map the extent of the thumb and index finger representation within SI and MI. Two local injections of 1 microl each (5 microg/microl) of the GABA(A)-agonist muscimol were used to inactivate the thumb and index region of either the pre- or post-central gyrus. The precision grip was differently affected by muscimol injection into either SI or MI. MI injections produced a deficit in the monkey's ability to perform independent finger movements and a general weakness in the finger muscles. Whole-hand grasping movements were inappropriately performed in an attempt to grasp either the instrumented object or morsels of food. Although the effect seemed strongest on intrinsic hand muscles, a clear deficit in digit extension was also noted. As a result, the monkey was unable to lift and maintain the object within the position window for the required 2 s, and, over time, the grip force decreased progressively until the animal stopped working. Following SI injections, the most obvious effect was a loss of finger coordination. In grasping, the placement of the fingers on the object was often abnormal and the monkey seemed unable to control the application of prehensile and lifting forces. However, the detailed analysis of forces revealed that a substantial increase in the grip force occurred well before any deficit in the coordination of finger movements was noted. This observation suggests that cutaneous feedback to SI is essential for the fine control of grip forces. PMID- 10473738 TI - Effects of unilateral brain damage on grip selection, coordination, and kinematics of ipsilesional prehension. AB - To determine whether the left and right hemispheres play specific roles in goal directed movements, prehension with the ipsilesional hand was tested in patients with unilateral brain damage. The task required that subjects rotate the hand while reaching for a bar that was presented in different orientations in the frontal plane, thus making high demands on visuospatial processing. The grasped bar had to be put into a hole: under one task condition the placement of the bar was specified, while under another it was not. The constrained task required that the subject anticipate the placing action when planning the initial prehensile movement. Grip selection, reaction times, kinematics of the transport movement, and coordination of hand rotation during transport were assessed in ipsilesional movements of 22 patients with either left or right brain damage (LBD and RBD) and in control subjects. Patients in both groups exhibited performance deficits; however, impairment characteristics differed profoundly between the groups. RBD patients showed prolonged reaction time and degraded kinematics in the unconstrained task, whereas LBD patients performed relatively well when only the orientation of the bar varied, but slowly and frequently incoordinated when the subsequent action was specified. Our findings emphasize the dominant role of the right hemisphere in processing visuospatial aspects of goal-directed movements, whereas the left hemisphere subserves non-spatial aspects of preplanning under increased task demands. Correlations of the patient's performance with results from clinical tests showed that neither deficits in visuospatial perception of RBD patients nor apraxia of LBD patients could account for the observed abnormalities in the use of the ipsilesional hand. PMID- 10473739 TI - Integration of the hand in postural reactions to sustained sideways force at the pelvis. AB - In order to investigate the potential hand contribution to sideways balance, sideways pushes to the right, which subjects resisted using either the lower limbs ("hip only") or the lower limbs assisted by the right upper limb ("hand and hip"), were delivered to the pelvis. Analysis of force and electromyogram recordings from the legs, arm and hand in the hand-and-hip condition showed a close co-ordination of upper and lower limbs in terms of mean latencies and amplitudes. However, trial to trial fluctuations of forces generated by the hand and leg did not correlate, suggesting parallel pathways under central co ordinative control. PMID- 10473740 TI - Effects of chronic median nerve compression at the wrist on sensation and manual skills. AB - The aim of this study was to analyse the functional impairments caused by chronic median nerve compression at the wrist on hand sensation and manual skill. Hand function was assessed in 11 patients (8 women and 3 men) with severe carpal tunnel syndrome (CTS) and compared with that of an age- and sex-matched control group. Apart from CTS, the subjects were healthy and the electrodiagnostic examination was normal. The pressure and vibration detection thresholds of the index finger were partially impaired and statistically different (P<0.05) when compared with controls, suggesting a reduction of tactile acuity in the territory of the median nerve. The thermal thresholds were identical in both groups, suggesting that the small-diameter fibres were not affected. When a small object was lifted and positioned in space, the coordination between the grip force and the vertical lifting force did not seem to be affected in our patients. They were able to modify their grip force according to the friction between the fingertips and the object, i.e. the more slippery the object, the higher the grip force. The unimanual Purdue Pegboard subtest results suggest that digital dexterity was also not significantly perturbed in our sample of CTS patients when compared with controls. Despite the severe abnormalities of median nerve conduction, our results suggest that chronic median nerve compression occurring in CTS induces partial impairment of tactile sensibility with minor impact on grasp force regulation and digital dexterity. PMID- 10473741 TI - Prehension movements and motor development in children. AB - The maturation of manual dexterity and other sensorimotor functions was assessed with various behavioural tests. In healthy children (age 4-5 years) and in adults, the kinematics of reaching and grasping, a bimanual task and fast repetitive tapping movements were analysed. Furthermore a comprehensive motor function score (MOT), probing agility and balance, was evaluated. In the prehension task, the straightness of the reaching trajectories increased with age. Children opened their grip relatively wider than adults, thus grasping with a higher safety margin. The speed of both tapping and bimanual movements increased with age, and higher scores were reached in the MOT. Although the different behavioural tests sensitively indicated maturational changes, their results were generally not correlated, i.e. the outcome of a particular test could not predict the results of other tasks. Hence there is no simple and uniform relationship between different behavioural data describing maturation of sensorimotor functions. PMID- 10473742 TI - Anticipatory grip adjustments are observed in both goal-directed movements and movement tics in an individual with Tourette's syndrome. AB - We examined grip force adjustments during movements of a hand-held object in a young man (BF) with Tourette's syndrome. We directly compared BF's voluntary up and down movements with tics in the same directions. Movement tics were elicited by cueing BF to move either up or down on a GO signal which appeared after a variable delay. During the delay period, we observed frequent tics which were almost always in the cued movement direction. BF's voluntary movements were well coordinated and featured precise and appropriate anticipatory grip force adjustments such that grip force was modulated in phase with movement-induced fluctuations in load. Precise anticipatory grip force adjustments were also observed in all of BF's movement tics. These results support the hypothesis that tics in Tourette's syndrome are purposeful voluntary movements that are well organized and coordinated. PMID- 10473744 TI - Impaired anticipatory finger grip-force adjustments in a case of cerebellar degeneration. AB - We describe adjustments in grip force as a consequence of fluctuations in inertial load force during vertical movements of the upper limb in a patient with cerebellar degeneration. Normally grip force is adapted to load-force fluctuations, in particular to the maximum load force, which occurs early in upward movements and late in downward movements. Increased grip force during movement was observed in the patient, but the timing of maximum grip force was not different between upward and downward movements. This suggests impaired cerebellar prediction of the dynamic consequences of voluntary movement. PMID- 10473743 TI - Grip-load force coordination in cerebellar patients. AB - The study examined the anticipatory grip force modulations to load force changes during a drawer-opening task. An impact force was induced by a mechanical stop which abruptly arrested movement of the pulling hand. In performing this task, normal subjects generated a typical grip force profile characterized by an initial force impulse related to drawer movement onset, followed by a ramp-like grip force increase prior to the impending load perturbation. Finally, a reactive response was triggered by the impact. In patients with bilateral cerebellar dysfunction, the drawer-opening task was performed with an alternative control strategy. During pulling, grip force was increased to a high (overestimated) default level. The latter suggests that cerebellar patients were unable to adjust and to scale precisely the grip force according to the load force. In addition, the latency between impact and reactive activity was prolonged in the patients, suggesting an impaired cerebellar transmission of the long-latency responses. In conclusion, these data demonstrate the involvement of cerebellar circuits in both proactive and reactive mechanisms in view of predictable load perturbations during manipulative behavior. PMID- 10473745 TI - Grip force dynamics in the approach to a collision. AB - This experiment investigated the prediction of load force (LF) in impulsive collisions inferred from anticipatory adjustments of grip force (GF) used to stabilise a hand-held object. Subjects used a precision grip to hold the object between thumb and index finger of their right hand and used the arm either: (1) to move the object to produce a collision by hitting the lower end of a pendulum, causing it to swing to one of three target angles, or (2) to hold the object still while receiving a collision produced by the experimenter releasing the pendulum from one of three angles. Visual feedback of the pendulum's trajectory was available in the production task only. In all conditions, subjects increased GF in advance of the collision. In receiving the collision without advance information, subjects set GF levels to the mid-range of the experienced forces. When subjects possessed knowledge about the maximum angle of pendulum swing - either because they were going to produce it or because they were verbally informed - magnitude of the anticipatory-GF magnitude response was scaled to the predicted LF magnitude. Furthermore, GF was scaled to LF with a higher gain when producing compared to receiving the collision. This suggests that updating forward models through a semantic route is not as powerful as when the updating is achieved through the more direct route of dynamic exploration. PMID- 10473746 TI - Coordination of reaching and grasping by capitalizing on obstacle avoidance and other constraints. AB - Reaching and grasping an object can be viewed as the solution of a multiple constraint satisfaction problem. The constraints include contact with the object with the appropriate effectors in the correct positions as well as generation of a collision-free trajectory. We have developed a computational model that simulates reaching and grasping based on these notions. The model, rendered as an animation program, reproduces many basic features of the kinematics of human reaching and grasping behavior. The core assumptions of the model are: (1) tasks are defined by flexibly organized constraint hierarchies; (2) manual positioning acts, including prehension acts, are first specified with respect to goal postures and then are specified with respect to movements towards those goal postures; (3) goal postures are found by identifying the stored posture that is most promising for the task, as determined by the constraint hierarchy, and then by generating postures that are more and more dissimilar to the most-promising stored posture until a deadline is reached, at which time the best posture that was found during the search is defined as the goal posture; (4) depending on when the best posture was encountered in the search, the deadline for the search in the next trial is either increased or decreased; (5) specification of a movement to the goal posture begins with straight-line interpolation in joint space between the starting posture and goal posture; (6) if an internal simulation of this default movement suggests that it will result in collision with an obstacle, the movement can be reshaped until an acceptable movement is found or until time runs out; (7) movement reshaping occurs by identifying a via posture that serves as a body position to which the actor moves from the starting posture and then back to the starting posture, while simultaneously making the main movement from the starting posture to the goal posture; (8) the via posture is identified using the same posture-generating algorithm as used to identify the goal posture. These processes are used both for arm positioning and, with some elaboration, for prehension. The model solves a number of problems with an earlier model, although it leaves some other problems unresolved. PMID- 10473747 TI - Parallel pathways mediating manual dexterity in the macaque. AB - Transmission of information along appropriately structured parallel pathways ensures that a great deal of information can be transferred from the source to the target very quickly, and with great security-essential features of any motor control system. Studies over the last two decades have established that the corticospinal and corticocerebellar pathways mediating manual dexterity in the primate are structurally organized to sustain the parallel transmission of sensorimotor information in multiple pathways. Serial, hierarchical control systems now seem insufficient to regulate voluntary hand movements. To achieve the required coordination, and precision and speed of execution, they must be combined with parallel control systems, which themselves incorporate elaborate feedforward and feedback controls. To illustrate these issues, two aspects of the structural organization of parallel sensorimotor pathways mediating manual dexterity in the macaque are reviewed. First, we examine the structure of the multiple corticospinal neuron subpopulations projecting from different areas of the frontoparietal cortex and how they are modified following hemisection of the cervical spinal cord. The remarkable recovery of hand function following spinal hemisection, despite the absence of any structural 'bridging' of the interrupted spinal pathways, and the fact that this is accountable in a parallel but not in a purely serial transmission system, are then reviewed. The second aspect of parallel distributed transmission examined is its occurrence within a single population of relay neurons. Our recent structural analysis of the somatic/dendritic organization of rubrospinal neurons in macaque red nucleus is used. The very large dendritic fields of individual neurons, extending over one third or more of the nucleus, provide a framework for extracting precise somatotopic information from an input population whose axon terminal arbors overlap extensively, and, which, without effective filtering, would provide poor spatial resolution. PMID- 10473748 TI - The role of synchrony and oscillations in the motor output. AB - There is currently much interest in the synchronisation of neural discharge and the potential role it may play in information coding within the nervous system. We describe some recent results from investigations of synchronisation within the motor system. Local field potentials (LFPs) and identified pyramidal tract neurones (PTNs) were recorded from the primary motor cortex of monkeys trained to perform a precision grip task. The LFPs showed bursts of oscillatory activity at 20-30 Hz, which were coherent with the rectified electromyographs (EMG) of contralateral hand and forearm muscles. This oscillatory synchronisation showed a highly specific task dependence, being present only during the part of the task when the animal maintained a steady grip and not during the movement phases before or after it. PTNs were phase-locked to LFP oscillations, implying that at least part of the coherence between cortical activity and EMG was mediated by corticospinal fibres. The phase locking of the PTNs to LFP oscillations produced task-dependent oscillatory synchronisation between PTN pairs, as assessed by the single-unit cross-correlation histogram. Recordings were also made from normal human subjects performing a precision grip similar to that used in the monkey recordings. Pairs of EMGs recorded from intrinsic hand and forearm muscles showed 20-30 Hz coherence, which modulated during task performance, being present only during periods of steady contraction. We suggest that these changes in EMG-EMG synchronisation reflect changing levels of synchronous drive from the corticospinal system. The generation of oscillations in the cortex is discussed in the light of results from a model of local cortical circuits. Other modelling work has shown that synchrony in the corticospinal inputs could act to recruit motoneurones more efficiently, producing more output force from a muscle than asynchronous inputs firing at the same mean rate. A speculative hypothesis is presented on the role of synchronous oscillations in the motor system, which is consistent with experimental observations to date. PMID- 10473749 TI - Precise spike synchronization in monkey motor cortex involved in preparation for movement. AB - It is commonly accepted that perceptually and behaviorally relevant events are reflected in changes of activity in largely distributed neuronal populations. However, it is much less clear how these populations organize dynamically to cope with momentary computational demands. In order to decipher the dynamic organization of cortical ensembles, the activities of up to seven neurons of the primary motor cortex were recorded simultaneously. A monkey was trained to perform a pointing task in six directions. During each trial, two signals were presented consecutively. The first signal provided prior information about the movement direction, whereas the second called for the execution of that movement. Dynamic interactions between the activity of simultaneously recorded neurons were studied by analyzing individual epochs of synchronized firing ("unitary events"). Unitary events were defined as synchronizations which occur significantly more often than expected by chance on the basis of the neurons' firing rates. The aim of the study was to describe the relationships between synchronization dynamics and changes in activity of the same neurons during the preparation and execution of voluntary movements. The data show that even neurons which were classified, on the basis of the change in their firing rate, to be functionally involved in different processes (e.g., preparation or execution related, different directional tuning) synchronized their spiking activity significantly. These findings indicate that the synchronization of individual action potentials and the modulation of the firing rate may serve different and complementary functions underlying the cortical organization of cognitive motor processes. PMID- 10473750 TI - Context-dependent force coding in motor and premotor cortical areas. AB - In three monkeys trained to finely grade grip force in a visuomotor step-tracking task, the effect of the context on neuronal force correlates was quantitatively assessed. Three trial types, which differed in force range, number, and direction of the force steps, were presented pseudo-randomly and cued with the color of the cursor serving as feedback of the exerted force. Quantitative analyses were made on 85 neurons with similar discharge patterns in the three trial types and significant linear positive (54 cells) or negative (31 cells) correlation coefficients between firing rate and force. An analysis of covariance (ANCOVA) showed that the population slopes for 2-step were steeper than for 3-step trials. Another ANCOVA at the population level, computed on the differences in firing rate and force between force steps, persistently disclosed a significant effect of trial type. For the first two force steps, the differences in firing rate were significantly larger in the 2-step than in the 3-step increase trials. Further analyses revealed that neither the force range nor the number of steps was a unique factor. A small group of neurons was tested in an additional trial series with a uniform cue for all three trials, leading to either a loss of context dependency or to unexpected changes in firing rate. This demonstrates that the cue color was an important instruction for task performance and neuronal activity. The most important findings are that the context-dependent changes were occurring "on-line", and that neurons displaying context-dependency were found in all three lateral premotor cortex hand regions and in the primary motor cortex. Finger muscle activity did not show any context dependency. The context-dependent effect leads to a normalization of the cortical activity. The advantage of normalization is discussed and mechanisms for the gain regulation are proposed. PMID- 10473751 TI - Learning, plasticity, and recovery in the central nervous system. AB - Cerebral functions can be described by the interaction of different brain regions as parts of distributed networks. Learning is seen as a refinement of the connection between the various parts of these networks. Plastic changes, as illustrated in brain charting techniques, are the result of learning (or use) in normal brains or found as adaptation (active or passive) after peripheral or central lesions. The relation between brain reorganization and recovery of function is investigated by two recent studies relating the training-induced improvement of lost function to changes in the brain. Others search for the effects of passive stimulation and drug influences. Independently of the approach, however, the general idea is that recovery can be seen as a reconnection between the remaining parts of the disturbed network. PMID- 10473752 TI - Somatotopic gradients in the distributed organization of the human primary motor cortex hand area: evidence from small infarcts. AB - Nine cases of relatively selective hand weakness produced by stroke were analyzed to examine the degree to which representations of different fingers are segregated in the human primary motor cortex (M1). In five cases, all the digits were involved uniformly; in four cases the radial versus ulnar digits of the hand were involved differentially. No patient showed discrete involvement of a single digit, nor did any patient have greatest weakness in the index, middle or ring finger. These findings provide little evidence that each digit is represented in a separate cortical territory, but rather suggest that broadly overlapping gradients - with the radial digits somewhat more heavily represented laterally and the ulnar digits somewhat more heavily represented medially - are superimposed on an underlying organization in which control of each finger is distributed widely throughout the human M1 hand area. PMID- 10473753 TI - Mechanisms of recovery of dexterity following unilateral lesion of the sensorimotor cortex in adult monkeys. AB - The mechanisms of recovery of manual dexterity after unilateral lesion of the sensorimotor cortex in adult primates remain a matter of debate. It has been proposed that the cortical zone adjacent to the lesion may take over part of the function of the damaged cortex. To investigate further this possibility, two adult (4-5 years old) macaque monkeys were trained to perform a natural precision grip task to assess hand dexterity. Intracortical microstimulations (ICMS) were used to map the hand area in M1 on both hemispheres. Ibotenic acid was then injected intracortically to damage the representation in M1 of the preferred hand. Subsequent histological analysis indicated that the hand representation in M1 was indeed lesioned, but, due to a spread of ibotenic acid, the lesion encroached a significant extent of the hand representation in the primary somatosensory cortex. A few minutes after infusion of ibotenic acid, there was a complete loss of dexterity of the preferred hand, which lasted for 1-2 months. Later, a progressive functional recovery of the affected hand took place over a 3 to 4-month period, reaching a stable level corresponding to 30% of the pre lesion behavioral score. ICMS remapping, conducted nine months after the lesion, revealed that stimulation of the intact or lesioned M1 did not induce any visible movement of the recovered hand. The M1 hand representation on the intact hemisphere was similar to that observed before the lesion. Transient inactivation of the M1 hand/arm areas or of the dorsal and ventral premotor cortical areas (PM) on both hemispheres was undertaken by using microinjections of the GABA agonist muscimol. Inactivations of M1 had no effect. Inhibition of PM in the damaged hemisphere suppressed the recovered manual dexterity of the affected hand. These results suggest that PM plays a significant role in the incomplete functional recovery of hand dexterity following unilateral damage of the sensorimotor cortex in adult monkeys. PMID- 10473754 TI - Neural representation of three-dimensional features of manipulation objects with stereopsis. AB - In the first part of this article, we review our neurophysiological studies of the hand-manipulation-related neurons in the anterior part of the lateral bank of the intraparietal sulcus (area AIP). We describe the properties of visually responsive neurons in area AIP. Object-type visual-dominant neurons responded to the sight of objects and showed selectivity not only for simple geometrical shapes, but also for complex objects such as a knob-in-groove and a plate-in groove. Some of the object-type visual-dominant neurons showed selectivity for the orientation of the longitudinal axis or the plane (surface) of a plate or a ring. In the second part of this article, we review our study of binocular visual neurons in the caudal part of the lateral bank of the intraparietal sulcus (c-IPS area), in particular, of axis-orientation-selective (AOS) neurons and surface orientation-selective (SOS) neurons. AOS neurons preferred long and thin stimuli, were sensitive to binocular disparity, and tuned to the axis orientation in three dimensional (3D) space. SOS neurons preferred broad and flat stimuli and were tuned to the surface orientation in depth. Some SOS neurons responded to a square in a random dot stereogram (RDS) with orientation tuning, suggesting that they encode surface orientation from a disparity gradient. Others responded to solid figure stereograms with orientation disparity and/or width disparity. It was concluded that the c-IPS area is a higher center for stereopsis, which integrates various binocular disparity signals received from the V3 complex and other prestriate areas to represent the neural code for 3D features. It may send projections to the AIP area and contribute to visual adjustment of the shape of the handgrip and/or hand orientation for manipulation and grasping. Neurons of the AIP area may also receive monocular cues of depth from the ventral visual pathway to discriminate the 3D shape of the object of manipulation. PMID- 10473755 TI - Gaze effects in the cerebral cortex: reference frames for space coding and action. AB - Visual information is mapped with respect to the retina within the early stages of the visual cortex. On the other hand, the brain has to achieve a representation of object location in a coordinate system that matches the reference frame used by the motor cortex to code reaching movement in space. The mechanism of the necessary coordinate transformation between the different frames of reference from the visual to the motor system as well as its localization within the cerebral cortex is still unclear. Coordinate transformation is traditionally described as a series of elementary computations along the visuomotor cortical pathways, and the motor system is thought to receive target information in a body-centered reference frame. However, neurons along these pathways have a number of similar properties and receive common input signals, suggesting that a non-retinocentric representation of object location in space might be available for sensory and motor purposes throughout the visuomotor pathway. This paper reviews recent findings showing that elementary input signals, such as retinal and eye position signals, reach the dorsal premotor cortex. We will also compare eye position effects in the premotor cortex with those described in the posterior parietal cortex. Our main thesis is that appropriate sensory input signals are distributed across the visuomotor continuum, and could potentially allow, in parallel, the emergence of multiple and task-dependent reference frames. PMID- 10473757 TI - Parcellation of the lateral premotor cortex of the macaque monkey based on staining with the neurofilament antibody SMI-32. AB - In macaque monkey, frontal and parasagittal brain sections were stained with SMI 32, an antibody directed against a nonphosphorylated neurofilament protein that labels pyramidal cells. The goal of this investigation was to find reliable criteria with which to draw the border between the motor (M1) and premotor (PM) cortex and delineate subdivisions within the lateral PM. Two-dimensional reconstruction of the staining patterns was also performed by flattening the series of frontal sections. The distribution of SMI-32 immunoreactivity in layers III and V of the cortex revealed the existence of three subregions in the ventral rostral PM and a clear mediolateral boundary within the dorsal PM defined by clusters of SMI-32-positive pyramidal cells in layer V. The border between M1 and PM was easily distinguished at the level of the dorsal PM by a strong loss of immunoreactive pyramidal cells in layers III and V. At the level of the ventral PM there was no clear disruption of layer V pattern, and the border was set using the pattern of layer III immunoreactivity. PMID- 10473756 TI - Largely segregated parietofrontal connections linking rostral intraparietal cortex (areas AIP and VIP) and the ventral premotor cortex (areas F5 and F4). AB - Two functionally different cortical areas are located in the rostral part of the intraparietal sulcus (IP): the ventral intraparietal area (VIP), along the fundus of the sulcus, and the anterior intraparietal area (AIP), rostral in the lateral bank. VIP and AIP have functional properties comparable to those of the ventral premotor areas, F4 and F5, respectively. The aim of this study was to establish whether these intraparietal and premotor areas have direct and specific anatomical connections. Neural tracers were injected in F4, F5, and AIP in three macaque monkeys. The results showed that F4 and F5 are targets of strong projections from VIP and AIP, respectively, and that the linkage between F5 and AIP is highly selective. These data support the notion that parietofrontal connections selectively link areas displaying similar functional properties and form largely segregated anatomical circuits. Each of these circuits is possibly dedicated to specific aspects of sensorimotor transformations. In particular, the AIP-F5 circuit should play a crucial role in visuomotor transformation for grasping, the VIP-F4 circuit is possibly involved in peripersonal space coding for movement. PMID- 10473758 TI - Visual responses in the dorsal premotor area F2 of the macaque monkey. AB - This study aimed to determine the presence of neurons responding to visual stimuli in area F2 of the dorsal premotor cortex of the macaque monkey. In order to delimit the sector in which visually responsive neurons are located, the somatotopic organization of area F2 was studied with intracortical microstimulation and single neuron recording. The results showed that: (1) in area F2 there is a significant percentage of visually responsive neurons (15.9% of all recorded neurons); (2) area F2 is excitable with a low-threshold current (average 28.1 microA) and has a somatotopic representation of the whole body, except the face; and (3) most visually driven neurons (n=130 out of 169) are concentrated within the rostrolateral sector of the forelimb representation of area F2, thus providing for the first time functional support for the neuroanatomical evidence that the visual input to area F2 is mostly restricted to this sector. PMID- 10473759 TI - Eye-hand coordination in uni- and bimanual goal-oriented tasks. AB - Two different drawer tasks were investigated with the aim of assessing the role of eye movements in well-coordinated hand movements. In an unimanual step tracking task, which had a predictive and an unpredictive movement, a two-way repeated-measures ANOVA showed a significant effect of prediction on the onset of grip-force (GF) rate (300+/-39 ms for the predictive condition versus 394+/-53 ms for the non-predictive condition, P<0.0001). Correlation coefficients, computed from the eye and the hand movements were low for the right and the left hand. The saccade was more coupled with the visual step change than with the action of the hand per se. In a second bimanual pull-and-pick task, the instruction was to pull a drawer with the left hand from a closed position to a LED-cued open position and then to grasp and reinsert a small peg in the drawer with the right hand. Correlation coefficients, computed from the latencies of saccades and of the leading left hand or of the right hand, were significant in four of five subjects. Intermanual correlations were significant in all five subjects. In conclusion, we found that the initial saccade in the unimanual task was best related with the visual step change, but was poorly correlated with the pulling/pushing hand. In the bimanual task, a moderate, but significant temporal coupling between the eyes and hand events was observed. This coupling was, however, less tight than that between both hands. PMID- 10473760 TI - Neuronal activity related to eye-hand coordination in the primate premotor cortex. AB - To test the functional implications of gaze signals that we previously reported in the dorsal premotor cortex (PMd), we trained two rhesus monkeys to point to visual targets presented on a touch screen while controlling their gaze orientation. Each monkey had to perform four different tasks. To initiate a trial, the monkey had to put his hand on a starting position at the center of the touch screen and fixate a fixation point. In one task, the animal had to make a reaching movement to a peripheral target randomly presented at one of eight possible locations on a circle while maintaining fixation at the center of this virtual circle (central fixation + reaching). In the second task, the monkey maintained fixation at the location of the upcoming peripheral target and, later, reached to that location. After a delay, the target was turned on and the monkey made a reaching arm movement (target fixation + reaching). In the third task, the monkey made a saccade to the target without any arm movement (saccade). Finally, in the fourth task, the monkey first made a saccade to the target, then reached to it after a delay (saccade + reaching). This design allowed us to examine the contribution of the oculomotor context to arm-related neuronal activity in PMd. We analyzed the effects of the task type on neuronal activity and found that many cells showed a task effect during the signal (26/60; 43%), set (16/49; 33%) and/or movement (15/54; 28%) epochs, depending on the oculomotor history. These findings, together with previously published data, suggest that PMd codes limb movement direction in a gaze-dependent manner and may, thus, play an important role in the brain mechanisms of eye-hand coordination during visually guided reaching. PMID- 10473761 TI - A parieto-premotor network for object manipulation: evidence from neuroimaging. AB - Functional magnetic resonance imaging (fMRI) was used to assess cerebral activation during manipulation of various complex meaningless objects as compared to manipulation of a single simple object (a sphere). Significant activation was found bilaterally in the ventral premotor cortex (Brodmann's area 44), in the cortex lining the anterior part of the intraparietal sulcus (most probably corresponding to monkey anterior intraparietal area, AIP), in the superior parietal lobule and in the opercular parietal cortex including the secondary somatosensory area (SII). We suggest that the cortex lining the anterior part of the intraparietal sulcus and area 44 are functionally connected and mediate object manipulation in humans. PMID- 10473762 TI - Time constraints improve reaching movements in an ataxic patient. AB - We report on a patient (AM) with a post-traumatic ataxia who has uncoordinated reaching movements to resting targets, but is able to catch moving objects. AM participated in three experiments to identify factors responsible for the favorable effect of object motion on her performance. In the first experiment, the task was to catch an object that moved away from AM. The speed of the object to be grasped (target object) varied. In experiment 2, the effect of time constraints on reaching performance was examined. AM had to reach for and grasp a stationary object and was allowed either 600 ms or 2000 ms to perform the task. In the third experiment, liquid crystal shutter glasses were used to manipulate the time that the subject was able to view the stationary target object and her reaching movements (vision-on time). While increased speed of the object, tighter time constraints, and short vision-on time hardly affected the performance of AM's unaffected left hand, they greatly improved her right-hand performance. These results are discussed in light of the hypothesis that the brain mechanisms controlling externally triggered movements differ from those controlling internally regulated movements. PMID- 10473763 TI - Do patients with neglect show abnormal hand velocity profiles during tactile exploration of peripersonal space? AB - It has been suggested that the movement impairments experienced by patients with neglect are not restricted to spatial disorders, but also affect higher-order kinematics (velocity and acceleration) to the extent that movements towards the neglected side are slower than movements away from it. In a recent study, we could not confirm this hypothesis, but found that patients with unilateral neglect exhibited no distinct direction-specific deficits in hand velocity when performing goal-directed reaching movements. Here we investigated whether neglect patients might reveal direction-specific deficits during exploratory hand movements. Six patients with left-sided neglect and six age-matched healthy control subjects scanned with their right hands the surface of a large table searching for a (non-existent) tactile target. Movements were performed in darkness. Time-position data of the hand were recorded with an optoelectronic camera system. Median activity of the patients' exploratory hand movements was shifted to the right (Karnath and Perenin 1998). Hand trajectories were partitioned into sections of leftward/rightward or, along the sagittal plane, into sections of near/far movements. For each movement section average and peak velocities were computed. The patients' hand movements were bradykinetic when compared with the control group. However, we found no evidence that average or peak velocities of leftward intervals were systematically lower than during rightward motion. Direction-specific deficits in velocity were also not observed for movements to and away from the body (sagittal plane). In conclusion, we found evidence for general bradykinesia in neglect patients but not for a direction specific deficit in the control of hand velocity during exploratory hand movements. PMID- 10473765 TI - Acquisition of anticipatory postural adjustments in a bimanual load-lifting task: normal and pathological aspects. AB - Anticipatory adjustments of forearm posture are associated with a voluntary load lifting movement in bimanual load-lifting tasks. Three aspects of these adjustments are analyzed: their goal, their central organization, and their acquisition. The goal of the anticipatory adjustment in this task is to minimize the perturbation of forearm posture that occurs during unloading. The central organization is based on two parallel controls responsible, respectively, for the lifting movement of the moving forearm and the anticipatory postural adjustment of the postural forearm, their coordination depending on a central timing signal. The acquisition of the anticipatory postural adjustment was tested using a paradigm where the voluntary movement performed by one hand triggered, via an electronic switch, the load release of the postural forearm. It was achieved after 40-60 trials and was not graded as a function of the voluntary movement parameters, but of the disturbance of the postural arm about to occur. The learned anticipation was not transferred when, after a first acquisition session with one forearm as the postural forearm, a second learning session was performed with the other forearm as the postural forearm. The acquisition was tested in Parkinsonian and in hemiparetic patients with capsular lesion. The highest acquisition deficit was observed in hemiparetic patients, when the contralateral forearm was the postural forearm; the deficit was less important when the ipsilateral arm was postural. Surprisingly, the anticipatory postural adjustments in hemiparetic patients were rather well preserved when the natural load-lifting task was tested. These results suggest that the basal-ganglia SMA circuit and M1 premotor areas are important in the acquisition process. PMID- 10473764 TI - Visual control of automated handwriting movements. AB - This study provides new insight into the subtle interaction between visual feedback and automated handwriting movements. To separate effects of visual feedback from effects of speed/accuracy trade-off, subjects were trained to perform the tasks with rapid and automated movements. To control the visual feedback the written trace was presented only on a computer screen. In the first experiment the visual feedback of script size was manipulated. Subjects had to write the character combination "ll" onto a pattern of squares. In some trials the script size was unpredictably enlarged by 133% or reduced by 66%. Results showed that the ongoing "l" was not affected by the manipulation, but subjects adapted script size in the following "l" without any change in the kinematic characteristic of the movements. In a second experiment trajectory correction strategies in perturbed writing movements were studied. Two small boxes had to be connected by rapid and single-stroke movements. In some of the trials the position of the target box was changed unpredictably to a new position. Kinematic analysis revealed that the initial movement was not aborted, but rather a second independent movement was added to correct the trajectory to the new target. Thus, this proves that a distortion of visual feedback does not directly slow down open loop movements to allow control of the motor output in a closed-loop mode. The ballisticity and automation was maintained during movement correction. Our findings fit perfectly well with recent theories of kinematic organization where complex movements are composed by elemental movement strokes. PMID- 10473766 TI - Role of the feedforward command and reafferent information in the coordination of a passing prehension task. AB - The performances of a deafferented patient and five control subjects have been studied during a self-driven passing task in which one hand has to grasp an object transported by the other hand and in a unimanual reach-to-grasp task. The kinematics of the reach and grasp components and the scaling of the grip aperture recorded for the self-driven passing task were very similar in controls and the deafferented subject (GL). In contrast, for the unimanual task when vision was absent, GL's coordination between reaching and grasping was delayed in space and time compared with the control subjects. In addition, frequent reopening of the grip was observed in GL during the final closure phase of the unimanual prehension task. These results support the notion that afferent proprioceptive information resulting from the reaching movement - which seemed to be used to coordinate reaching and grasping commands in the unimanual task - is no longer necessary in the self-induced passing task. Finally, for the externally driven passing task, when the object was passively transported by the experimenter, the coordination was consistently modified in all subjects; grip aperture onset was delayed, thus asserting a specific contribution of the central command or feedforward mechanisms into the anticipation of the grasp onset observed in the self-driven passing task. The origin and nature of the information necessary for building up the feedforward mechanisms remains to be elucidated. PMID- 10473767 TI - Cerebral midline structures in bimanual coordination. AB - In six healthy right-handed volunteers, we compared the cerebral activation pattern related to unimanual right- and left-hand movements and to bimanual in phase and anti-phase movements using functional magnetic resonance imaging (fMRI). Internally paced unimanual finger-to-thumb opposition movements led to a strong contralateral activation of primary sensorimotor areas in all six subjects. Midline activity was lateralized to the left side during right-hand movements, but to both sides during left-hand movements. Activity patterns of bimanual in-phase movements resembled the combined activity patterns of the two unimanual conditions: right and left hemispheric activations of the primary sensorimotor cortices and predominantly left-sided medial frontal activity. In contrast, during anti-phase movements, we observed a clear increase in activity, in both right and left frontal midline areas and in right hemispheric, mainly dorsolateral premotor areas compared to in-phase movements. These results indicate that frontal midline activity is not specific for bimanual movements per se. It can already be involved during simple unimanual movements but becomes progressively more involved during more complex aspects of movement control. PMID- 10473768 TI - The identification of coordination constraints across planes of motion. AB - Two dominant coordination constraints have been identified during isofrequency conditions in previous work: the egocentric constraint, i.e., simultaneous activation of homologous muscle groups, and the allocentric constraint, i.e., moving the segments in the same direction in extrinsic space. To verify their generalization, bimanual drawing movements were performed in different planes of motion (transverse, frontal, sagittal, frontal-transverse) according to the in phase and anti-phase mode along the X- and Y-axes. Convergent findings were obtained across the transverse, frontal, and frontal-transverse planes. The in phase mode along both axes was performed most accurately/consistently, whereas the anti-phase mode resulted in a deterioration of the coordination pattern and this effect was most pronounced when the latter mode was introduced with respect to both dimensions. For sagittal plane motions, the in-phase mode was again superior but the second most optimal configuration was the anti-phase mode along both axes. This finding was hypothesized to result from the familiarity with the pattern since it resembles cycling behavior. It illustrates how cognitive mapping is superimposed onto the dynamics of interlimb coordination. Overall, these results support the presence of both the egocentric and allocentric constraint during bimanual movement production. PMID- 10473769 TI - Evidence for interhemispheric motor-level transfer in a simple reaction time task: an EEG study. AB - Simple visuomanual reaction time tasks require interhemispheric communication when stimuli are presented in the hemifield opposite the responding hand. Although confirmed in many studies, it is still a matter of debate when, at what functional level and at what site this interhemispheric transfer takes place. To address these questions, we recorded event-related potentials (ERPs) in 12 healthy subjects performing such a task and analyzed the data using techniques based on topographic ERP map characteristics. A method which has proved useful for associating ERP map configurations of different time periods with functional states of the brain was supplemented by a source localization procedure. The results suggest that transfer occurs late in time, on a functional motor level and at frontal sites, at least for left-to-right interhemispheric direction of transfer. PMID- 10473770 TI - Molecular evolution of helix-turn-helix proteins. AB - The helix-turn-helix domain-containing family of transcriptional regulators is of ancient origin and has been incorporated into numerous disparate biological processes. As a consequence, the forces shaping its early evolution have been difficult to reconstruct. Herein, we analyze this large and diverse family with a combination of traditional phylogenetic techniques and newer sequence analysis tools to determine whether the helix-turn-helix family arose from a single common ancestor. Our analyses of the DNA-binding domain show that amino acid chemistry is conserved at many sites in the first helix and the turn. The high level of divergence combined with the short length of the domain hinders robust reconstruction of the entire phylogeny, but some level of deep node inference is possible. All analyses point to a predominantly monophyletic origin for the helix turn-helix domain. The consequences of such an origin for a diverse group of proteins, and guidelines for the identification of future members of the HTH family are discussed. PMID- 10473771 TI - Molecular data from the chloroplast rpoC1 gene suggest a deep and distinct dichotomy of contemporary spermatophytes into two monophyla: gymnosperms (including Gnetales) and angiosperms. AB - Partial sequences of the rpoC1 gene from two species of angiosperms and three species of gymnosperms (8330 base pairs) were determined and compared. The data obtained support the hypothesis that angiosperms and gymnosperms are monophyletic and none of the recent groups of the latter is sister to angiosperms. PMID- 10473772 TI - Evolutionary history of MHC class I genes in the mammalian order Perissodactyla. AB - We carried out an analysis of partial sequences from expressed major histocompatibility complex (MHC) class I genes isolated from a range of equid species and more distantly related members of the mammalian order Perissodactyla. Phylogenetic analysis revealed a minimum of six groups, five of which contained genes and alleles that are found in equid species and one group specific to the rhinoceros. Four of the groups contained only one, or very few sequences, indicating the presence of relatively nonpolymorphic loci, while another group contained the majority of the equid sequences identified. These data suggest that a diversification of MHC genes took place after the split between the Equidae and the Rhinocerotidae yet before the speciation events within the genus Equus. PMID- 10473773 TI - Compositional correlations in the chicken genome. AB - This paper analyses the compositional correlations that hold in the chicken genome. Significant linear correlations were found among the regions studied coding sequences (and their first, second, and third codon positions), flanking regions (5' and 3'), and introns-as is the case in the human genome. We found that these compositional correlations are not limited to global GC levels but even extend to individual bases. Furthermore, an analysis of 1037 coding sequences has confirmed a correlation among GC(3), GC(2), and GC(1). The implications of these results are discussed. PMID- 10473774 TI - Synonymous and nonsynonymous substitutions in genes from Gramineae: intragenic correlations. AB - In this work, we have investigated the relationships between synonymous and nonsynonymous rates and base composition in coding sequences from Gramineae to analyze the factors underlying the variation in substitutional rates. We have shown that in these genes the rates of nucleotide divergence, both synonymous and nonsynonymous, are, to some extent, dependent on each other and on the base composition. In the first place, the variation in nonsynonymous rate is related to the GC level at the second codon position (the higher the GC(2) level, the higher the amino acid replacement rate). The correlation is especially strong with T(2), the coefficients being significant in the three data sets analyzed. This correlation between nonsynonymous rate and base composition at the second codon position is also detectable at the intragenic level, which implies that the factors that tend to increase the intergenic variance in nonsynonymous rates also affect the intragenic variance. On the other hand, we have shown that the synonymous rate is strongly correlated with the GC(3) level. This correlation is observed both across genes and at the intragenic level. Similarly, the nonsynonymous rate is also affected at the intragenic level by GC(3) level, like the silent rate. In fact, synonymous and nonsynonymous rates exhibit a parallel behavior in relation to GC(3) level, indicating that the intragenic patterns of both silent and amino acid divergence rates are influenced in a similar way by the intragenic variation of GC(3). This result, taken together with the fact that the number of genes displaying intragenic correlation coefficients between synonymous and nonsynonymous rates is not very high, but higher than random expectation (in the three data sets analyzed), strongly suggests that the processes of silent and amino acid replacement divergence are, at least in part, driven by common evolutionary forces in genes from Gramineae. PMID- 10473775 TI - Recent selection on synonymous codon usage in Drosophila. AB - Evidence from a variety of sources indicates that selection has influenced synonymous codon usage in Drosophila. It has generally been difficult, however, to distinguish selection that acted in the distant past from ongoing selection. However, under a neutral model, polymorphisms usually reflect more recent mutations than fixed differences between species and may, therefore, be useful for inferring recent selection. If the ancestral state is preferred, selection should shift the frequency distribution of derived states/site toward lower values; if the ancestral is unpreferred, selection should increase the number of derived states/site. Polymorphisms were classified as ancestrally preferred or unpreferred for several genes of D. simulans and D. melanogaster. A computer simulation of coalescence was employed to derive the expected frequency distributions of derived states/site under various modifications of the Wright Fisher neutral model, and distributions of test statistics (t and Mann-Whitney U) were derived by appropriate sampling. One-tailed tests were applied to transformed frequency data to assess whether the two frequency distributions deviated from neutral expectations in the direction predicted by selection on codon usage. Several genes from D. simulans appear to be subject to recent selection on synonymous codons, including one gene with low codon bias, esterase 6. Selection may also be acting in D. melanogaster. PMID- 10473776 TI - The role of interelement selection in Saccharomyces cerevisiae Ty element evolution. AB - Retrotransposons are mobile genetic elements that are ubiquitous components of eukaryotic genomes. The evolutionary success of retrotransposons is explained by their ability to replicate faster than the host genomes in which they reside. Elements with higher rates of genomic replication possess a selective advantage over less active elements. Retrotransposon populations, therefore, are shaped largely by selective forces acting at the genomic level between elements. To evaluate rigorously the effects of selective forces acting on retrotransposons, detailed information on the patterns of molecular variation within and between retrotransposon families is needed. The sequencing of the Saccharomyces cerevisiae genome, which includes the entire genomic complement of yeast retrotransposons, provides an unprecedented opportunity to access and analyze such data. In this study, we analyzed in detail the patterns of nucleotide variation within the open reading frames of two parental (Ty1 and Ty2) and one hybrid (Ty1/2) family of yeast retrotransposons. The pattern and distribution of nucleotide changes on the phylogenetic reconstructions of the three families of Ty elements reveal evidence of negative selection on both internal and external branches of the Ty phylogenies. These results indicate that most, if not all, Ty elements examined represent active or recently active retrotransposon lineages. We discuss the relevance of these findings with respect to the coevolutionary dynamic operating between genomic element populations and the host organisms in which they reside. PMID- 10473777 TI - Identification of multiple Gypsy LTR-retrotransposon lineages in vertebrate genomes. AB - Gypsy LTR-retrotransposons have been identified in the genomes of many organisms, but only a small number of vertebrate examples have been reported to date. Here we show that members of this family are likely to be widespread in many vertebrate classes with the possible exceptions of mammals and birds. Phylogenetic analyses demonstrate that although there are several distinct lineages of vertebrate gypsy LTR-retrotransposons, the majority clusters into one monophyletic clade. Groups of fungal, plant, and insect elements were also observed, suggesting horizontal transfer between phyla may be infrequent. However, in contrast to this, there was little evidence to support sister relationships between elements derived from vertebrate and insect hosts. In fact, the majority of the vertebrate elements appeared to be most closely related to a group of gypsy LTR-retrotransposons present within fungi. This implies either that at least one horizontal transmission between these two phyla has occurred previously or that a gypsy LTR-retrotransposon lineage has been lost from insect taxa. PMID- 10473778 TI - Speciation versus phenotypic plasticity in coral inhabiting barnacles: Darwin's observations in an ecological context. AB - Speciation and phenotypic plasticity are two extreme strategic modes enabling a given taxon to populate a broad ecological niche. One of the organismal models which stimulated Darwin's ideas on speciation was the Cirripedia (barnacles), to which he dedicated a large monograph. In several cases, including the coral inhabiting barnacle genera Savignium and Cantellius (formerly Pyrgoma and Creusia, respectively), Darwin assigned barnacle specimens to morphological "varieties" (as opposed to species) within a genus. Despite having been the subject of taxonomic investigations and revisions ever since, the significance of these varieties has never been examined with respect to host-associated speciation processes. Here we provide evidence from molecular (12S mt rDNA sequences) and micromorphological (SEM) studies, suggesting that these closely related barnacle genera utilize opposite strategies for populating a suite of live-coral substrates. Cantellius demonstrates a relatively low genetic variability, despite inhabiting a wide range of corals. The species C. pallidus alone was found on three coral families, belonging to distinct higher-order classification units. In contrast, Savignium barnacles exhibit large between- and within-species variations with respect to both micromorphology and DNA sequences, with S. dentatum "varieties" clustering phylogenetically according to their coral host species (all of which are members of a single family). Thus, whereas Savignium seems to have undergone intense host-associated speciation over a relatively narrow taxonomic range of hosts, Cantellius shows phenotypic plasticity over a much larger range. This dichotomy correlates with differences in life-history parameters between these barnacle taxa, including host infestation characteristics, reproductive strategies, and larval trophic type. PMID- 10473779 TI - Intron-genome size relationship on a large evolutionary scale. AB - The intron-genome size relationship was studied across a wide evolutionary range (from slime mold and yeast to human and maize), as well as the relationship between genome size and the ratio of intervening/coding sequence size. The average intron size is scaled to genome size with a slope of about one-fourth for the log-transformed values; i.e., on the global scale its increase in evolution is lower than the increase in genome size by four orders of magnitude. There are exceptions to the general trend. In baker's yeast introns are extraordinarily long for its genome size. Tetrapods also have longer introns than expected for their genome sizes. In teleost fish the mean intron size does not differ significantly, notwithstanding the differences in genome size. In contrast to previous reports, avian introns were not found to be significantly shorter than introns of mammals, although avian genomes are smaller than genomes of mammals on average by about a factor of 2.5. The extra-/intragenic ratio of noncoding DNA can be higher in fungi than in animals, notwithstanding the smaller fungal genomes. In vertebrates and invertebrates taken separately, this ratio is increasing as the increase in genome size. Two hypotheses are proposed to explain the variation in the extra-/intragenic ratio of noncoding DNA in organisms with similar numbers of genes: transition (dynamic) and equilibrium (static). According to the transition model, this variation arises with the rapid shift of genome size because the bulk of extragenic DNA can be changed more rapidly than the finely interspersed intron sequences. The equilibrium model assumes that this variation is a result of selective adjustment of genome size with constraints imposed on the intron size due to its putative link to chromatin structure (and constraints of the splicing machinery). PMID- 10473780 TI - Molecular clock calibrations and metazoan divergence dates. AB - It has recently been argued that living metazoans diverged over 800 million years ago, based on evidence from 22 nuclear genes for such a deep divergence between vertebrates and arthropods (Gu 1998). Two "internal" calibration points were used. However, only one fossil divergence date (the mammal-bird split) was directly used to calibrate the molecular clock. The second calibration point (the primate-rodent split) was based on molecular estimates that were ultimately also calibrated by the same mammal-bird split. However, the first tetrapods that can be assigned with confidence to either the mammal (synapsid) lineage or the bird (diapsid) lineage are approximately 288 million years old, while the first mammals that can be assigned with confidence to either the primate or the rodent lineages are 65 million years old, or 85 million years old if ferungulates are part of the primate lineage and zhelestids are accepted as ferungulate relatives. Recalibration of the protein data using these fossil dates indicates that metazoans diverged between 791 and 528 million years ago, a result broadly consistent with the palaeontological documentation of the "Cambrian explosion." The third, "external" calibration point (the metazoan-fungal divergence) was similarly problematic, since it was based on a controversial molecular study (which in turn used fossil dates including the mammal-bird split); direct use of fossils for this calibration point gives the absurd dating of 455 million years for metazoan divergences. Similar calibration problems affect another recent study (Wang et al. 1999), which proposes divergences for metazoans of 1000 million years or more: recalibrations of their clock again yields much more recent dates, some consistent with a "Cambrian explosion" scenario. Molecular clock studies have persuasively argued for the imperfection of the fossil record but have rarely acknowledged that their inferences are also directly based on this same record. PMID- 10473781 TI - Branch length heterogeneity leads to nonindependent branch length estimates and can decrease the efficiency of methods of phylogenetic inference. AB - Branch length estimates play a central role in maximum-likelihood (ML) and minimum-evolution (ME) methods of phylogenetic inference. For various reasons, branch length estimates are not statistically independent under ML or ME. We studied the response of correlations among branch length estimates to the degree of among-branch length heterogeneity (BLH) in the model (true) tree. The frequency and magnitude of (especially negative) correlations among branch length estimates were both shown to increase as BLH increases under simulation and analytically. For ML, we used the correct model (Jukes-Cantor). For ME, we employed ordinary least-squares (OLS) branch lengths estimated under both simple p-distances and Jukes-Cantor distances, analyzed with and without an among-site rate heterogeneity parameter. The efficiency of ME and ML was also shown to decrease in response to increased BLH. We note that the shape of the true tree will in part determine BLH and represents a critical factor in the probability of recovering the correct topology. An important finding suggests that researchers cannot expect that different branches that were in fact the same length will have the same probability of being accurately reconstructed when BLH exists in the overall tree. We conclude that methods designed to minimize the interdependencies of branch length estimates (BLEs) may (1) reduce both the variance and the covariance associated with the estimates and (2) increase the efficiency of model based optimality criteria. We speculate on possible ways to reduce the nonindependence of BLEs under OLS and ML. PMID- 10473782 TI - The period gene: high conservation of the region coding for Thr-Gly dipeptides in the Drosophila nasuta species subgroup. AB - In this study, the region corresponding to the Thr-Gly region of the period (per) gene in the Drosophila nasuta subgroup of species was sequenced. The results showed that this region was highly conserved in the D. nasuta subgroup. There were only nine variable sites found in this 300-bp-long region, all located in two small regions highly variable among Drosophila species. No length variation was observed either within this subgroup or in the Yunnan (YN) population of D. albomicans. The deduced amino acid sequences were identical for all 14 taxa in the D. nasuta subgroup, and a stretch of alternating Thr-Gly pairs was not observed in this subgroup. A phylogenetic tree was constructed. The clustering of some species was in general agreement with previous works, but it also raised some question on the phylogenetic relationship between the nasuta species. The data did not implicate the Thr-Gly region playing a role in behavioral isolation in this subgroup of Drosophila. PMID- 10473783 TI - Oxime reactivation of RBC acetylcholinesterases for biomonitoring. AB - A low-variability method to reactivate blood cholinesterases (ChEs) after prior exposure of mammals, including humans, to ChE-inhibiting organophosphate esters (OPs) is presented. A concentration of 10 mM pyridine 2-aldoxime methochloride (2 PAM Cl) was incubated with intact red blood cells (RBCs) and assayed virtually free of interfering oxime and hemoglobin (Hb). Variability was decreased by reducing the number of washing steps and sedimenting RBC ghosts through a 7% sucrose cushion. Statistically significant detections of reactivations as low as 5% with average "false positives" of 3.8% were achieved. Relative rates and extent of reactivation after OP treatment of rabbit RBC AChE in vitro were of the order dimethyl- (DDVP) > diethyl- (ethyl paraoxon) >, diisopropyl-substituted (diisopropyl fluorophosphate; DFP) OPs. Rabbit RBC AChE was reactivatable for up to 60 h following dermal exposure to ethyl parathion and reactivatable for only 12 to 24 h following exposure to methyl parathion. Reactivation of plasma ChEs with 0.1 mM 2-PAM Cl in the same animals was achievable for only 12 to 24 h after ethyl parathion and for only 1 to 4 h after methyl parathion. PMID- 10473784 TI - Polychlorinated Dibenzo-p-Dioxins and Dibenzofurans in Storm Water Outfalls Adjacent to Urban Areas and Petroleum Refineries in San Francisco Bay, California. AB - Polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) were analyzed in storm water collected from 15 outfalls in San Francisco Bay, California, considered representative of areas located adjacent to petroleum refineries and mixed urban/commercial/residential land uses. Storm water sampling was conducted hourly over a 12-h period at two outfalls located near the city of Oakland and the suburban community of Benicia, which represent urban and mixed urban/light industrial land uses, respectively. Sampling was conducted hourly to determine temporal changes in the concentrations and distributions of PCDD/Fs during the first major rainstorm event of the 1995/96 winter season. In addition, storm water sampling was conducted in December 1995 and February/April 1996 at six publicly owned treatment work (POTW) outfalls located adjacent to developed, undeveloped, mixed residential/commercial, and industrial lands. Storm water sampling also was conducted at five outfalls located adjacent to petroleum refineries. The sampling results were used to examine changes in PCDD/F loadings to San Francisco Bay at the onset and midway through the winter rainy season and to characterize differences in PCDD/F concentrations, if any, between refinery and nonrefinery areas. The concentrations of tetra- through octa-chlorinated homologues and 2,3,7,8-substituted congeners were assayed in filtered storm water samples according to US EPA Method 1613A. Principal components analysis (PCA) was used as an exploratory technique to examine the similarities and differences in the distributions of PCDD/Fs in storm water from different outfalls. The sampling results showed few significant differences between storm water discharged from outfalls located in mixed urban/commercial/residential areas and outfalls located adjacent to petroleum refineries. The concentrations of 2,3,7,8-TCDD were below the analytical limits of detection in all storm water samples, with the exception of samples collected after the sixth hour of sampling at the Oakland outfall. Fingerprint patterns were generally dominated by the higher chlorinated PCDD/F congeners including OCDD, OCDF, and 1,2,3,4,6,7,8-HpCDD. Total TEQs were generally below US EPA ambient water quality criteria and aquatic ecotoxicology guideline values. The results of this study strongly suggest that discrete sampling of storm water outfalls is insufficient to characterize the concentrations and distributions of persistent hydrophobic contaminants such as PCDD/Fs. In addition to surface water runoff from the city of Oakland, other nonindustrialized urban locations may represent important sources of PCDD/Fs to San Francisco Bay.http://link.springer ny.com/link/service/journals/00244/bibs/37n3p290.html PMID- 10473785 TI - Persistent Organochlorines in the Effluents from a Chlorine-Bleached Kraft Integrated Pulp and Paper Mill in Southeast Asia. AB - A bleached sulfate integrated pulp and paper mill producing printing and writing paper from mixed tropical hardwood and bamboo was studied. The mill uses a "conventional bleaching sequence," C-E-H1-H2, with an average molecular chlorine consumption of 50 kg per ton of air-dried pulp (ADP). The content of polychlorinated dibenzofurans (PCDFs) and dibenzo-p-dioxins (PCDDs) in the bleaching filtrate in terms of the nordic toxicity equivalent (N-TEQ) was 33.5, 1.15, 0.56, and 0.014 pg/L for the E, C, H1, and H2 bleaching stages, respectively. The corresponding PCDFs and PCDDs loads in ng/t ADP were in the same ranking, i.e., 670, 69, 11.2, and 0.28, respectively. The congener and isomeric pattern of PCDFs and PCDDs of the bleaching filtrate and the bleached pulp was found to be typical for the chlorine bleaching plant effluent. The obtained dioxin load formed in the mill is in agreement with Western studies for the given multiple chlorine of 0.21-0.23. The load is, however, lower than reported discharges from Scandinavian mills using 1980s bleaching technologies, but substantially higher than the discharges from mills with modern bleaching technologies. Modifications in the bleaching plant to reduce molecular chlorine use are necessary to reduce dioxin formation.http://link.springer ny.com/link/service/journals/00244/bibs/37n3p303.html PMID- 10473786 TI - Sorption of Nine Pesticides to Three Aquatic Macrophytes. AB - The sorption of nine pesticides to the aquatic macrophytes Chara globularis, Elodea nuttallii, and Lemna gibba was studied. A batch equilibrium method was used to study the sorption at five concentration levels to fresh shoots of the macrophytes. The results for the herbicides atrazine and linuron were described by nonlinear Freundlich equations, with Freundlich exponents ranging from 0.53 to 0.60. The results for the other compounds showed almost linear sorption isotherms, with Freundlich exponents ranging from 0.9 to 1.1. The highest sorption was measured for chlorpyrifos, with sorption coefficients ranging from 1,660 to 2,150 L/kg. Sorption coefficients for C. globularis tended to be lower than those for the other two macrophytes. Correlation (R(2) = 0.80) was found for the relation between the sorption coefficient (K(d)) of six pesticides and their solubility in water (S). The equation log K(d) = 3.20 - 0.65 log S can be used for a first estimate of the sorption coefficient of a pesticide to aquatic macrophytes.http://link.springer ny.com/link/service/journals/00244/bibs/37n3p310.html PMID- 10473787 TI - Polychlorinated biphenyl- and mercury-associated alterations on benthic invertebrate community structure in a contaminated salt marsh in southeast Georgia. AB - The community structure of a benthic macroinvertebrate assemblage in a contaminated salt marsh was evaluated as part of an ecological characterization of a former chloralkali production facility in Georgia. Sample locations were chosen based on a gradient of the primary contaminants of concern, total mercury and polychlorinated biphenyls (PCBs), primarily Aroclor 1268. Sediment concentrations of Aroclor 1268 ranged from 2.3 to 150 mg/kg dry weight, while mercury concentrations ranged from 15 to 170 mg/kg dry weight in the study area. Mercury and PCBs were determined to be co-located in the sediments. Total organic carbon composition of the sediments was negatively associated with PCB and mercury concentrations. A total of 29 benthic taxa was identified in 49 samples; replicate samples were taken at each of five sampling locations. Mean infaunal density across all sampling locations was estimated at approximately 61,000 to 234,000 organisms m(-2). Overall, polychaetes comprised 57% of the infaunal community with Manayunkia aestuarina as the dominant species. Oligochaetes, nematodes, crustacea, insects, and gastropods comprised 23.0, 18.0, 1.0, 0.7, and 0.2% of the overall benthic community, respectively. Density estimates of individual species between sampling locations showed no consistent patterns in response to pollutants. However, an analysis of higher taxonomic levels revealed some general trends. In uncontaminated areas, the benthic community was dominated by nematodes and oligochaetes, whereas moderate to highly contaminated areas were dominated by polychaetes and a smaller percentage of oligochaetes and nematodes. A trophic analysis of the same data set revealed that the community shifted from an evenly distributed percentage of surface and subsurface feeders in the uncontaminated areas to a community dominated by surface feeders in the more contaminated locations. Carnivores comprised from 0.13 to 0.90% of the trophic structure, with the percentage of carnivores generally decreasing with increasing contamination. Mercury and PCBs were bioaccumulating in representative marsh benthic invertebrates, presenting a potential source of contaminants to marsh consumers. Tissue PCB and tissue mercury concentrations were positively related to sediment PCB and mercury concentrations, respectively. A standard 14-day toxicity test using the amphipod Leptocheirus plumulosus showed no acute toxicity across the sampling locations. PMID- 10473788 TI - Comparing metal toxicity among Daphnia magna clones: an approach using concentration-time-response surfaces. AB - This study investigates the use of concentration-time-response surfaces as a tool to predict potential long-term effects of metals to Daphnia magna, using mortality as an endpoint. Specifically, concentration-time-response surfaces were determined for four D. magna clones exposed to four metals (Cd, Zn, Cu or U) in moderate-hard or hard synthetic freshwater for 24-96 h. Mortality data were log(e) transformed into probits and then regressed against the reciprocal of exposure time and concentration. The results obtained clearly showed that mortality was markedly affected at exposure periods longer than 48 h. Thus, an optimal exposure period of 72 h was selected to compare predicted lethality effects (LC) of different intensities (i.e., 10-50%) derived from concentration time-response surfaces with measured chronic lethal levels obtained from the literature. Only the results of Cd and Cu show good agreement between predicted and measured chronic lethal concentration levels. The apparent disagreement observed for U could be attributed to differences in water quality, and hence, U bioavailability. The high levels of Zn toxicity observed in relation to the predicted values could indicate that chronic mortality of Zn is mediated by toxic anorexia. Overall, the results obtained show that concentration-time-response surfaces offered the potential to assess the effect of time on toxicity, which is desirable to credibly extrapolate from acute to chronic scenarios. Furthermore, by determining lower mortality thresholds (i.e., LC(10)) at different times, concentration-time-response surfaces were able to emulate the mode of action of the selected metals on chronic exposures. Therefore, the use of concentration time-response surfaces has potential application in environmental risk assessment. PMID- 10473789 TI - Tolerance to Zinc in Populations of the Earthworm Lumbricus rubellus from Uncontaminated and Metal-Contaminated Ecosystems. AB - Zinc tolerance in Lumbricus rubellus populations from two metal-polluted (smelter and mine) sites was studied by comparing the effects of zinc with responses in a reference site strain. For the study, adult worms were collected directly from the field. Thus, no attempt was made to differentiate between tolerance resulting from population-level genetic adaptation or phenotypic plasticity in metal physiology. To compare relative sensitivity for zinc, worms from the three populations were exposed in laboratory tests. Effects on survival, weight change, cocoon production, and internal zinc levels were measured. Prior to exposure, it was anticipated that worms from the metal-contaminated sites would show substantially increased tolerance to zinc. This was not the case for all measured parameters. Thus, although differences in the shape of the dose-response relationships for survival and cocoon production were found, substantial variations in measured responses, effect concentrations, or zinc accumulation rates were not apparent. Overall, therefore, zinc tolerance is unlikely to be a major factor influencing the distribution of L. rubellus in contaminated regions.http://link.springer ny.com/link/service/journals/00244/bibs/37n3p332.html PMID- 10473790 TI - Chronic Toxicity of Ammonia to New Zealand Freshwater Invertebrates: A Mesocosm Study. AB - Freshwater macroinvertebrate communities were established within 12 artificial streams or "toroidal" mesocosms and exposed to three replicated concentrations of ammonia for 29 days at constant temperature (16 degrees C) and pH (median 8.4). The criterion units (CU = measured [ammonia]/US EPA 1985 chronic criterion value) of total ammonia in the LOW, MED, and HIGH treatments were 2.0, 4.8, and 13 CUs respectively, and 1.9, 5.8, and 12 CUs for the unionized ammonia. Macroinvertebrates were tolerant of the ammonia exposures with no significant (p > 0.1) effect on taxa richness; number of taxa in the orders Ephemeroptera, Plecoptera, and Trichoptera (EPT); or the quantitative macroinvertebrate community index (QMCI), a biotic index proposed for assessing effects of organic enrichment in New Zealand streams. Significant differences (p < 0.05) occurred for the mean abundance and the numbers of EPT individuals (QEPT), with the HIGH treatment significantly lower (-41%) than the control for both abundance and QEPT. Of the major species, only the mayflies Deleatidium sp. (Ephemeroptera: Leptophlebiidae) and Coloburiscus humeralis (Ephemeroptera: Oligoneuriidae) showed significant reductions in abundance, with only the caddisflies Beraeoptera roria (Trichoptera: Conoesucidae) and Confluens sp. (Trichoptera: Conoesucidae) showing significant increases in abundance. The abundance of juvenile Deleatidium sp. had a negative concentration-response relationship that resulted in an 82% decrease in abundance in the HIGH treatment. Drift of invertebrates showed no response to ammonia treatments. The 29-day EC(50) values for Deleatidium sp. for total and unionized ammonia were 2.15 mg (N)/L (pH 8.4) and 0.145 mg (NH(3)-N)/L. No observed effect concentration (NOEC) values were 0.95 mg (N)/L and 0.066 mg (NH(3)-N)/L, and the threshold effect concentration (TEC) was 1.49 mg (N)/L and 0.102 mg (NH(3)-N)/L. Comparison of the Deleatidium sp. chronic ammonia sensitivity data with the US EPA 1985 chronic criterion value (CCC = 0.45 mg [N]/L, pH 8.4, 16 degrees C) showed the TEC value for total ammonia was 3.3x CCC, and 2.2x higher than the updated US EPA 1998 criteria. The findings suggest that use of the US EPA criteria would provide minimal protection for Deleatidium for chronic ammonia exposure, and that development of site-specific criteria, covering a wide range of environmental conditions, may be required to adequately protect this species.http://link.springer ny.com/link/service/journals/00244/bibs/37n3p338.html PMID- 10473791 TI - Uptake of particulate lead via the gills of fish (Carassius auratus). AB - Carassius auratus were exposed to mixtures of constant free lead (0. 1 mg/L) and particulate lead (adsorbed on gibbsite particles) at varied concentrations (0, 0.4, 0.8, 1.2, 2.0, and 3.0 mg/L). The concentrations of lead and aluminum in the gills and intestines of the fish were determined after exposure. The gills were examined (after the exposure) with a light microscope for any particles adhering to the surface. The examination results revealed that lead accumulations on the gills increased with increased particulate lead concentrations in the ambient water (under conditions of a constant level of free lead), indicating the bioavailability of lead via the gills. Gibbsite particles were also observed on the gill surface after exposure. The ratio of lead and aluminum in the gills was much higher than that in the particles. Our results suggest that there was a two step process involved in the uptake of particulate lead by fish gills: (1) adherence of the particles on the gill surface where mucus was attached; and (2) desorption of lead from the particles under conditions of the gill microenvironment. PMID- 10473792 TI - Cytological and biochemical alterations in Carassius auratus hepatocytes from exposure to sediment containing dioxins and related compounds. AB - Cytological and biochemical alterations of crucial carp (Carassius auratus) hepatocytes were characterized after exposure to sediments from a lake contaminated with dioxins and other industrial chemicals. Carp were exposed in 20 L water containing 25, 50, or 100 g of contaminated sediment for 2 and 4 weeks. Ultrastructural changes in the liver were characterized by severe enlargement of hepatocytes. Alterations in the cell included formation of condensed and irregular cell nucleus, polynuclei, dispersed heterochromatin, enlargement of the nucleolus, and degeneration of the nucleus. Mitochondrial numbers were reduced and cristae were deformed. Myelin figures and lysosomes were increased, and sometimes cell organelles and cell matrix were totally lost after 4 weeks of exposure. The ultrastructural alterations were correlated with exposure time and sediment concentrations. Hepatosometic index was significantly increased in experimental groups at 2 and 4 weeks as compared with the control group. EROD enzyme activities were strongly induced in liver. A trend from rough endoplasmic reticulum (RER) to SER was observed. Our results suggest that the dioxin-like compounds bound by sediment were bioavailable to C. auratus and cause sublethal effects. PMID- 10473794 TI - Effects of mercury on health and first-year survival of free-ranging great egrets (Ardea albus) from southern Florida. AB - The objectives of this study were to determine whether elevated mercury (Hg) concentrations have a negative impact on the health and survival of nestling and juvenile free-ranging great egrets (Ardea albus) from southern Florida. During 1994, when health and survival was monitored in a cohort of young birds with naturally variable concentrations of Hg, packed cell volume was positively correlated with blood Hg concentrations, and high Hg concentration in blood was not related to the probability of surviving during the first 10.5 months of life. During 1995, 70 first-hatched great egret chicks were included in a Hg field dosing experiment to compare the effects of elevated Hg on health and survival. Birds were dosed while in the nest orally every 2.5 days for 15 days with 0.5 mg of methyl mercury chloride (MeHgCl) for an estimated intake of 1.54 mg MeHgCl/kg food intake. These birds were compared with controls, which received an estimated 0.41 mg MeHgCl/kg food. No differences were observed in health parameters or in the probability of surviving during the first 8 months of age between egrets that were dosed with Hg and those that were not. A likely explanation for the lack of any effects on health and survival between both groups could be that chicks at this age were eliminating most of the dietary Hg through the production of new feathers. PMID- 10473793 TI - Relative distribution of polychlorinated biphenyls among tissues of neonatal American alligators (Alligator mississippiensis). AB - Recent papers have investigated the utility of chemical analysis of the chorioallantoic membrane (CAM) as a nonlethal indicator of avian and reptilian exposure to persistent environmental contaminants. This study was undertaken to evaluate the chemical distribution among fat, CAM, and residual yolk tissues of live neonatal American alligators (Alligator mississippiensis) to investigate the potential utility of CAM use as a nonlethal indicator of exposure to polychlorinated biphenyls (PCBs). CAMs were collected from neonatal alligators at hatch, whereas yolk sacs and fat tissues were taken from each animal at euthanization (3 weeks posthatch). All collected tissue samples were separately Soxhlet extracted and analyzed by GC/ECD for PCBs. Log normalized, individual PCB congener concentrations in CAMs were significantly correlated with concentrations in fat (r(2) = 0.62) and yolk (r(2) = 0.56) tissues. Coefficients of determination from comparisons of homologue group concentrations varied from 0.13 to 0.90. Residue levels in neonatal oviparous organism tissues may be qualitatively assessed through chemical analyses of CAM tissues removed from discarded eggshells. PMID- 10473795 TI - Cadmium and lead exposure associated with reduced growth rates, poorer fledging success of little blue heron chicks (Egretta caerulea) in south Louisiana wetlands. AB - Persistent contaminants in ecosystems are often monitored via organisms that bioaccumulate the pollutant of interest. Several published studies have used colonial wading birds to assess contamination by heavy metals in aquatic ecosystems. Using a nonlethal sampling regimen, this study identified chick feathers and guano as tissues that best reflect local contamination. Both cadmium and lead were detected in food samples (0.3-4.3 microg/g Cd, 0.8-9.3 microg/g Pb), guano (0.2-3.2 microg/g Cd, 1.0-9.5 microg/g Pb), and feathers (0.6-25.4 microg/g Cd, 1.2-16.9 microg/g Pb) from little blue heron chicks at levels similar to other studies. Chicks exposed to cadmium had significantly slower growth rates than nonexposed chicks, and exposure to lead was correlated with increased nestling mortality. The appropriate scale of comparison necessary to demonstrate these effects and the usefulness of wading birds as bioindicators are discussed. PMID- 10473796 TI - Metal concentrations in the eggs of endangered Florida scrub-jays from central Florida. AB - We analyzed metal levels in the eggs of the endangered Florida scrub-jay (Aphelocoma coerulescens), an extremely sedentary and modestly long-lived passerine bird. Eggs were obtained as part of the long-term study of this species at the Archbold Biological Station, located in south central Florida. Following a brief and usually short dispersal, Florida scrub-jays remain in a permanent territory in their scrub oak habitat, where they feed mainly on insects and acorns. As they are low on the food chain and sedentary, we expected levels of contaminants to be comparatively low and to reflect the local environment. No significant yearly differences existed for any metal between eggs collected in 1997 (n = 11) and 1998 (n = 5). Arithmetic mean metal concentrations (whole egg, dry weight) were low for arsenic (34 ppb), cadmium (25 ppb), mercury (74 ppb), and lead (66 ppb), intermediate for chromium (226 ppb), and higher for selenium (1,470 ppb) and manganese (2,230 ppb). Except for the low mercury value, all these means were within the range of mean values reported for a wide range of nonpasserine species, including those at high trophic levels, but are lower than concentrations associated with abnormalities in birds. The three essential elements (chromium, selenium, and manganese) were approximately normally distributed, whereas arsenic, cadmium, lead, and mercury were highly skewed (arithmetic mean much higher than geometric mean). PMID- 10473797 TI - Comparative study of cadmium transfer in ewe and cow milks during rennet and lactic curds preparation. AB - Cadmium transfer from whole milk to cream, rennet, or lactic curds was studied before and following a repeated oral cadmium administration to three lactating ewes and one cow. Before cadmium administration, the cadmium levels in milk were around 0.4 microg/L in ewes and less than 0.2 microg/L in cow. Throughout cadmium administration the mean cadmium levels in milk were 3.3+/-1.4 microg/L in ewes and 2.5+/-1 microg/L in cow. During cadmium administration, 86% of cadmium in ewe milk was dispersed in the skimmed milk and 17% in the cream, whereas only 72% was dispersed in the cow skimmed milk and 27% in the cow cream. Most of milk cadmium was associated with casein fractions. About 70% of milk cadmium was transferred to the rennet or lactic curds of ewe. The remaining cadmium present in whole milk, about 9%, was transferred to the rennet or lactic curd whey. In cow, the proportion of cadmium associated with rennet or lactic curds, rennet curd whey, and lactic curd whey was, respectively, 60%, 56%, 14% and 12% of total milk cadmium. The fraction of total cadmium transferred from milk to its milk products, whatever the species, ranged from 94% to 103%. The factor of concentration of cadmium from whole milk to milk products ranged from three to six. We suggest that the excretion of cadmium into milk is mainly achieved via the milk casein secretion. This is, to our knowledge, the first in vivo study where the cadmium transfer from milk to its milk products after repeated cadmium oral administration to ewe and cow has been studied. PMID- 10473798 TI - Contamination levels and specific accumulation of persistent organochlorines in Caspian seal (Phoca caspica) from the Caspian sea, Russia. AB - Persistent organochlorines, such as polychlorinated biphenyls (PCBs) including coplanar congeners, DDTs, HCHs, chlordanes (CHLs), and HCB, were determined in the blubber of Caspian seals (Phoca caspica) and their fish diet (Rutilus sp.) collected in 1993 from the northern Caspian Sea, Russia. Notable concentrations of DDTs and HCHs were found at mean values of 19 and 1.3 microg/g on wet-weight basis in adult male seals, respectively. PCB pollution in Caspian seals was not so considerable compared with those of seals that suffered mass mortality. Less gender difference of organochlorine residue levels in adult animals implies less excretion of organochlorines from the body of adult females through lactation and gestation, probably due to the higher rate of pregnancy failure. Immature seals had a wide range of organochlorine concentrations, which decreased as body length increased, suggesting dilution. Caspian seals can be considered to have higher degradation capacity for coplanar PCBs. Mean TEQs (2,3,7,8-TCDD toxic equivalents) for non-, mono- and di-ortho coplanar PCBs was 51 pg/g on wet-weight basis, which was lower than those in seals that have suffered mass mortality, but comparable to those found in Arctic seals. PMID- 10473799 TI - Distribution of PCB congeners, DDE, hexachlorobenzene, and methylsulfonyl metabolites of PCB and DDE among various fractions of human blood plasma. AB - The concentrations of chlorinated biphenyls (CBs), 1, 1-bis(4-chlorophenyl)-2,2 dichloroethene (DDE), hexachlorobenzene (HCB), and the methylsulfonyl metabolites of CBs (MeSO(2)-CBs) and DDE (MeSO(2)-DDE) were determined in human plasma samples and in the fractions obtained by ultracentrifugation of plasma into very low-density (VLDL), low-density (LDL), high-density (HDL) lipoprotein and lipoprotein depleted (LPDP) fractions (containing primarily albumin). The concentrations of triacylglycerols, cholesterol, phospholipids, and apolipoprotein B (apoB) were determined. The organochlorine compounds were associated with all fractions, but predominantly with the LPDP fraction. On an average 44% of CBs, 61% of MeSO(2)-CBs, 73% of DDE, 77% of MeSO(2)-DDE, and 45% of HCB were distributed in the LPDP fraction. A tendency to greater association of 3-methylsulfonyl substituted than of corresponding 4-methylsulfonyl substituted chlorobiphenyls to the LPDP fraction was noticed. Among the lipoprotein fractions, LDL was the main carrier of HCB, DDE and CBs. MeSO(2)-DDE was predominantly found in HDL and MeSO(2)-CBs were distributed equally among the LDL and HDL fractions. Calculating the concentrations of organochlorine compounds in relation to the content of apoB, the levels were about 10 times higher in VLDL than in LDL. PMID- 10473800 TI - Human semen quality in relation to dietary pesticide exposure and organic diet. AB - The objective of the study was to corroborate or refute the hypothesis that farmers having a high intake of organic grown commodities have a high semen quality due to their expected lower level of dietary pesticides intake. Food frequency data and semen were collected from 256 farmers (171 traditional farmers and 85 organic farmers, overall participation rate: 32%) who were selected from central registers. Each farmer delivered one semen sample before the spraying season started. The farmers were divided into three groups where the commodities from organic production contributed no (N, 0%), medium (M, 1-49%), or a high (H, 50-100%) proportion of the fruit and vegetables consumed. Farmers having a high relative intake of organically grown fruit and vegetables also had a high relative consumption of organically produced meat, milk, and bread, and differences were observed comparing the actual mean intake of single commodities, such as rice, potato, and pork meat. The current individual dietary intake of 40 pesticides was estimated using food frequencies and generalized serving size data in combination with data on pesticide concentrations in food commodities as obtained from the National Danish Food Monitoring Program. The estimated pesticide intake was significantly lower among farmers of group H, but for all three groups of farmers the average dietary intake of 40 pesticides was at or below 1% of the acceptable daily intake (ADI) except for the dithiocarbamates (max = 0.21 microg/kg day = 2.2% ADI), methidathion, (max = 0.01 microg/kg day = 1.4% ADI), and 2-phenylphenol (max = 0.21 microg/kg day = 1.1% ADI). The median sperm concentration for the three groups of farmers was not significantly different (p = 0.40, median sperm concentration was N = 62, M = 44, and H = 75 million/ml). The group of men without organic food intake had a significant lower proportion of morphologically normal spermatozoa, but in relation to 14 other semen parameters no significant differences were found between the groups. Intake of 40 individual pesticides was correlated with four semen parameters (concentration, percentage dead spermatozoa, percentage normal sperm heads, and motility [VCL]). Five significant correlations (p value 0.01) were found among the 160 comparisons in relation to percentage dead spermatozoa: azinphos-methyl, carbaryl, chlorfenson, fenitrothion, and tetradifon. For all of them a lower percentage of dead spermatozoa were found in the groups with a high dietary intake of the specific pesticide. In contrast, for all pesticides evaluated only minor differences were found between the groups when considering spermatozoa concentration, morphology, and motility. In conclusion, the estimated dietary intake of 40 pesticides did not entail a risk of impaired semen quality, but precautions should be taken when generalizing this negative result to populations with a higher dietary exposure level or an intake of other groups of pesticides. PMID- 10473802 TI - Generating a T cell tumor-specific immune response in vivo: can flt3-ligand generated dendritic cells tip the balance? AB - flt3 ligand (FL) is a growth factor that induces hematopoietic progenitor cell and dendritic cell (DC) expansion when administered to mice. Lymphoid-related (CD8alpha(+)) and myeloid-related (CD8alpha(-)) DC are transiently expanded in multiple tissues. Treatment of tumor-bearing mice with FL results in slower tumor growth and, in some cases, tumor rejection and the development of tumor-specific T cell immunity. The clinical use of DC as cellular vehicles for tumor antigen presentation to generate a tumor-specific T cell response is under investigation. DC are currently generated ex vivo, pulsed with antigen, and then infused into patients, and much effort is being directed toward optimizing each of these steps. Administration of FL to humans induces a profound increase in circulating DC. The availability of a large number of DC generated in vivo has important implications for tumor immunotherapy approaches. PMID- 10473803 TI - The induction of distinct cytokine cascades correlates with different effects of granulocyte-colony stimulating factor and granulocyte/macrophage-colony stimulating factor on the lymphocyte compartment in the course of high-dose chemotherapy for breast cancer. AB - The availability of the myeloid hemopoietic growth factors (HGF) granulocyte- and granulocyte/macrophage-colony stimulating factor (G-CSF and GM-CSF) has enhanced the therapeutic index of high-dose chemotherapeutic antitumoral regimens (HDCT), as well as the rate of severe damage to immune competence. We investigated some immune functions before, during and after one course of HDCT for poor-risk breast cancer and compared the effects of G-CSF and GM-CSF on the immune recovery. They exerted different influences on the functions we examined and showed distinctive patterns of both qualitative and quantitative in vivo activities on the immune system. The main findings were that (a) granulocyte and lymphocyte recovery rates were faster in the patients receiving G-CSF; (b) looking at the lymphocyte compartment, this difference was restricted to the CD3(+)/CD8(+) and CD56(+) lymphocyte subsets; (c) the reconstitution rate of CD19(+) lymphocytes was slow in both groups; (d) at the end of follow-up HLA-DR expression by CD3(+) lymphocytes was higher in the GM-CSF group; (e) the lymphocyte proliferative capacity was restored at a faster rate in the GM-CSF group, whereas cytotoxic activities recovered better in the G-CSF group; (f) the early repopulating phase was characterized by higher interleukin-6 serum levels in the GM-CSF group. Overall, GM-CSF seemed to exert an earlier effect on all T lymphocyte subsets, preventing them from a complete drop during the long-lasting "nadir" of the cell count, whereas G-CSF appeared to boost them strongly, though a few days later, hastening their final recovery. The distinct pattern of the cytokine cascade induced by each factor, consistent with the different functional changes, seemed to account for the peculiarities of their immune modulations. PMID- 10473804 TI - Synergistic antitumor effects of interleukin-12 gene transfer and systemic administration of interleukin-18 in a mouse bladder cancer model. AB - We introduced the interleukin-12 (IL-12) gene into the mouse bladder cancer cell line (MBT2) to establish sublines that secrete bioactive IL-12. IL-12-secreting MBT2 (MBT2/IL-12) sublines were completely rejected when subcutaneously implanted into immunocompetent syngeneic C3H mice. Although this antitumor effect did not change when IL-12-secreting cells were injected into immunodeficient mice whose CD8(+) T or CD4(+) T cells had been depleted by the corresponding antibody, it was abrogated when natural killer cells were depleted by anti-asialoGM1 antibody. In addition, when parental MBT2 cells mixed with MBT2/IL-12 cells were subcutaneously injected into mice, admixed MBT2/IL-12 inhibited the growth of the parental tumor. Furthermore, this antitumor effect was enhanced by systemic IL-18 administration. This synergism was abrogated when the mice were treated with interferon-gamma-neutralizing antibody in vivo. In conclusion, local secretion of IL-12 led to effective antitumor activity that was enhanced by systemic administration of IL-18. Interferon-gamma plays an important role in the synergism of IL-12 gene transduction and systemic administration of IL-18. PMID- 10473805 TI - An EGP-2/Ep-CAM-expressing transgenic rat model to evaluate antibody-mediated immunotherapy. AB - The human pancarcinoma-associated epithelial glycoprotein-2 (EGP-2), also known as 17-1A or Ep-CAM, is a 38-kDa transmembrane antigen, commonly used for targeted immunotherapy of carcinomas. Although strongly expressed by most carcinomas, EGP 2 is also expressed in most simple epithelia. To evaluate treatment-associated effects and side-effects on tumor and normal tissue respectively, we generated an EGP-2-expressing transgenic Wistar rat. To express the cDNA of the EGP-2 in an epithelium-specific manner, the 5' and 3' distal flanking regions of the human keratin 18 (K18) gene were used. EGP-2 protein expression was observed in the liver and pancreas, whereas EGP-2 mRNA could also be detected in lung, intestine, stomach and kidney tissues. In this rat, EGP-2-positive tumors can be induced by injecting a rat-derived carcinoma cell line transfected with the GA733-2 cDNA encoding EGP-2. Transgenic rats were used to study specific in vivo localization of an i.v. anti-EGP-2 monoclonal antibody, MOC31, applied i.v. Immunohistochemical analyses showed the specific localization of MOC31 in s.c. induced EGP-2-positive tumors, as well as in the liver. In contrast, in EGP-2 transgenic rats, MOC31 did not bind to EGP-2-negative tumors, the pancreas, or other normal tissues in vivo. In conclusion, an EGP-2-transgenic rat model has been generated that serves as a model to evaluate the efficacy and safety of a variety of anti-EGP-2-based immunotherapeutic modalities. PMID- 10473806 TI - ImmTher, a lipophilic disaccharide derivative of muramyl dipeptide, up-regulates specific monocyte cytokine genes and activates monocyte-mediated tumoricidal activity. AB - ImmTher, a liposome-encapsulated lipophilic disaccharide tripeptide derivative of muramyl dipeptide, previously showed activity against liver and lung colorectal metastases in a phase I trial. The purpose of the current studies was to investigate whether ImmTher could up-regulate specific cytokine gene expression and protein production, as well as activate the tumoricidal or cytostatic activity of human monocytes. ImmTher induced the expression and production of interleukin(IL)-1alpha IL-1beta, IL-6, IL-8, IL-12, macrophage chemotactic and activating factor, and tumor necrosis factor alpha but not IL-2 or IL-10. Cytostatic or cytotoxic monocyte activity was stimulated against human Ewing's sarcoma, osteosarcoma, and melanoma cells but not breast cancer cells. Production and secretion of these cytokine proteins may play a role in the antitumor activity of ImmTher. PMID- 10473807 TI - Gene therapy with cytokine-transfected xenogenic cells (Vero-IL-2) in patients with metastatic solid tumors: mechanism(s) of elimination of the transgene carrying cells. AB - Eleven patients with advanced cancer were treated in a clinical gene therapy trial by repeated intra- tumoral injections with different doses of xenogenic fibroblasts secreting high amounts of human interleukin-2 (Vero-IL2). Treatments in a total of 14 courses were well tolerated and resulted in clinical responses and measurable biological effects. Together with increases in serum interleukin-2 (IL-2), modifications of the V-beta T cell receptor repertoire and induction of intratumoral T-cell infiltration were observed. When the intratumoral expression of endogenous cytokine genes and the persistence of the IL-2 transgene at the application site and in peripheral blood were investigated, rapid disappearance of the transgene at the application site appeared to be the most prominent biological effect. Tests detecting a single Vero-IL2 cell against a background of 10(5) non-transfected cells were not able to demonstrate significant expression of exogenous IL-2 (i.e. the transgene or transgene-carrying cells) in tumor biopsies or blood at different times. Therefore, further studies were performed to evaluate the mechanism(s) involved in the rapid disappearance of xenogenic carrier cells in more detail. We show here that significant in vitro cytotoxicity against transgene-carrying Vero cells can be observed in peripheral blood of all the patients before treatment as well as in healthy controls. "Cold" target inhibition shows that significant killing of Vero-IL2 cells is mediated by natural killer (NK) cells. This was confirmed by showing that established CD3( )/CD16(+)/CD56(+) peripheral blood NK cell clones kill both K562 and Vero-IL2 target cells. The failure of other mechanisms (complement, antibody-dependent cell cytotoxicity or cytotoxic T lymphocytes) to destroy xenogenic, histoincompatible Vero cells in vitro suggests that NK cells also might be responsible for the killing of Vero-IL2 in vivo and for the failure to detect the transgene at the application site. These results might also be of importance for some aspects of the current discussion of xenotransplantation. PMID- 10473808 TI - The role of possible risk factors for acute and late renal dysfunction after high dose interleukin-2, interferon alpha and lymphokine-activated killer cells. AB - Renal dysfunction is a frequently encountered adverse event following treatment with high-dose interleukin-2 (IL-2). Information about parameters predicting the severity of IL-2-associated renal function abnormalities is limited. In this study the role of possible risk factors in the development of high-dose IL-2 associated acute and long-term renal dysfunction was investigated. A total of 72 patients, who were treated with a regimen consisting of IL-2 (18 MIU m(-2) day( 1) by continuous infusion), interferon alpha (IFNalpha; 5 MIU m(-2) day(-1), intramuscularly) and lymphokine-activated killer (LAK) lymphocytes, were analysed. Serum creatinine measurements were performed daily during treatment, weekly between courses and monthly during follow-up. Pre-and posttreatment 24-h urine samples were collected for calculation of creatinine clearances. Renal dysfunction was observed in 97% of patients. Grade 1 dysfunction (according to WHO criteria) was observed in 20 patients (28%), grade 2 in 37 (51%), grade 3 in 13 (14%) and grade 4 in 0 (0%). Renal dysfunction was reversible in more than 90% of patients. Only 6 patients (8%) suffered a certain amount of permanent function loss. More severe acute renal dysfunction occurred in patients who were experiencing hypertension prior to treatment, those who suffered sepsis during treatment and in men than in women. Sepsis was also associated with irreversible function loss. Other variables such as age, performance status, diabetes mellitus, interval between nephrectomy and start of IL-2 therapy and hypotension during treatment were not important. In conclusion, with high-dose IL-2, renal dysfunction develops in almost every patient and such abnormalities are mostly reversible. Predictors for severe acute renal dysfunction are pre-existing hypertension, sepsis and sex. A septic episode also carries a risk of permanent damage. PMID- 10473809 TI - Generation of protective immunity against an immunogenic carcinoma requires CD40/CD40L and B7/CD28 interactions but not CD4(+) T cells. AB - Interactions between CD40 and CD40L play a central role in the regulation of both humoral and cellular immunity. Recently, interactions between these molecules have also been implicated in the generation of protective cell-mediated tumor immunity. We have generated a tumor model in which a well-understood and clearly immunostimulatory antigen, influenza hemagglutinin has been transfected into the BALB/c-derived, MHC-class-I-positive, B7-deficient murine mammary carcinoma, MT901. In this model, expression of the influenza hemagglutinin antigen does not alter tumorigenicity in naive but serves as a tumor-rejection target in immunized mice. T-cell-depletion experiments indicate that successful tumor protection can occur following immunization in mice depleted of CD4(+) but not CD8(+) T cells, suggesting that tumor protection is largely CD8-mediated and CD4-independent. Interestingly, despite the ability of tumor protection to be generated in the absence of CD4(+) T cells, effective immunization was clearly dependent on CD40/CD40L as well as CD28/B7 interactions. PMID- 10473823 TI - Glutamate toxicity induced degeneration of outer hair cells with a temporal increase of nitric oxide production in the guinea pig cochlea. AB - The aim of this study was to examine the roles of glutamate (GLU) toxicity and involvement of nitric oxide (NO) in the pathogenesis of cochlear degeneration. We examined guinea pig cochleae following chronic exposure to GLU. Trypan blue extrusion and transmission electron microscopy were performed to evaluate degeneration in the organ of Corti. In parallel, nitric oxide synthase (NOS) activity was demonstrated by histochemical staining of NADPH diapholase. GLU treatment caused time-dependent degeneration of outer hair cells (OHCs) in conjunction with a temporal increase of NOS activity in the organ of Corti. This suggests that GLU may be involved in OHC degeneration under toxic conditions, with NO production possibly playing a role in this process. PMID- 10473824 TI - Allergic fungal sinusitis caused by Bipolaris (Drechslera) hawaiiensis. AB - Depending on the aggressiveness of the pathogen and a patient's immunocompetence, fungal polypoid pansinusitis or allergic fungal sinusitis (AFS) may be a life threatening disease. Apart from the clinical findings, its diagnosis is based on the demonstration of mucinous material with abundant eosinophils in the paranasal sinuses (indicating an allergic process), cultivation of the causative pathogen and immunocompetence of the patient. In a 20-year-old immigrant Sudanese woman, AFS due to Bipolaris (Drechslera) hawaiiensis was diagnosed. Because of intracranial extension, the disease had led to erosion of the cranial base and orbit with amaurosis on the right side and focal epilepsy. In addition to endonasal microsurgical pansinus operations, local irrigation therapy with amphothericin B was accompanied by systemic treatment with itraconazole after in vitro cultivation of the pathogen and determination of its sensitivities. Interdisciplinary management included a combination of endonasal surgery with debridement of infected tissues and wide drainage of the sinuses without removal of skull bone or the dural lesion in addition to specific antimycotic treatment. Injury to adjacent anatomical structures must be avoided in any case to prevent systemic or possibly lethal dissemination of infection. PMID- 10473825 TI - Alterations of nasal mucociliary transport in patients with hypertrophy of the inferior turbinates, deviations of the nasal septum and chronic sinusitis. AB - Mucociliary transport (MCT) represents the first barrier of the nasal fossae and paranasal sinuses against various biological and physical insults. We studied the nasal MCT time using a mixture of vegetable charcoal powder and 3% saccharin in three groups of patients suffering from hypertrophy of the inferior turbinates, deviations of the nasal septum or chronic sinusitis. The mean values of the nasal MCT in the first two groups were practically identical to the normal ones. In contrast, significantly delayed times were found in patients with chronic sinusitis (P < 0.01). Findings indicate that this delay is determined by an increase in viscoelasticity of the mucus following the acute release of mediators of inflammation, together with a reduction in the periciliary stratum, which slows down the metachronous wave of the MCT. PMID- 10473826 TI - Effect of cromolyn pre-treatment on capsaicin-induced rhinitis in rats. AB - The effect of cromolyn sodium local pre-treatment on capsaicin-induced rhinitis in rats was studied by analyzing tissue changes due to edema, inflammatory cell infiltration and IgA upregulation. Nasal mucosa samples were stained with hematoxylin-eosin and examined immunohistochemically with monoclonal IgA antibodies. Changes were analyzed at 6, 12 and 72 h after capsaicin treatment and were scored semiquantitatively. Results showed that local cromolyn pre-treatment modified all parameters observed in the nasal mucosa following capsaicin-induced rhinitis in the rats. PMID- 10473827 TI - Optic nerve decompression in trauma and tumor patients. AB - Optic nerve decompression is a procedure that is now receiving increasing clinical attention. However, there are currently no standardized treatment protocols in the therapy of traumatic or pressure insults to the nerve. The present retrospective study was designed to report our experience with microscopic endonasal transethmoid-sphenoid optic nerve decompression in 24 unilateral trauma cases and 11 unilateral skull base tumor patients. In general preoperative visual acuities in the trauma patients were worse than in the tumor patients. Following surgery, 9 of 11 tumor patients (82%) had at least some improvement of their vision, including 5 complete recoveries. In the group with traumatic visual impairment, 16 of the patients had no light perception preoperatively. Postoperatively, 13 patients (54%) had at least some improvement, with 4 patients regaining normal or near normal vision. Compared to other techniques and approaches, our technique is a minimally invasive procedure for optic nerve decompression, reducing unnecessary operative trauma to nasal structures, skin incisions or even craniotomy and frontal lobe retraction. PMID- 10473828 TI - Proteolytic patterns of head and neck squamous cell carcinoma. AB - The significance of plasminogen activators and matrix metalloproteases for clinical outcome, growth and metastatic behavior of head and neck squamous cell carcinoma (SCC) is still controversial. The majority of studies has been based on either immunohistological stainings, which provide only limited quantitative information, or in vitro experiments. We analyzed 44 head and neck SCC and 11 mucosa tissue samples for the expression of gelatinolytic or fibrinolytic proteases by quantitative zymographic analysis and compared lytic activities to clinical and histopathological data. We calculated activation ratios for matrix metalloproteinases-2 and -9 (MMP-2 and MMP-9) by separate evaluations of inactive and activated MMP forms. Increased gelatinolytic and fibrinolytic activity was found in head and neck SCC when compared to mucosa. Increased values were caused by MMP-9 and urokinase type plasminogen activator, respectively. No statistically significant correlations of either protease lytic activity or activation ratio could be related to T-stage, metastasis, tissue necrosis or the differentiation stage of tumors. The data recorded are compared with previously published reports. PMID- 10473829 TI - Long-term results of different treatment modalities in 37 patients with glomus jugulare tumors. AB - The results of different forms of treatment of 37 patients with previously untreated glomus jugulare tumors were compared retrospectively. According to the Fisch classification system, 6 patients presented with class B tumors, 19 class C and 12 patients with class D. Twenty-eight patients underwent surgery and 9 patients had primary radiation therapy (to 50 Gy). In 20 of the surgical cases (71%), radical tumor removal could be achieved and required no further treatment over a follow-up period of 8.6 years (range 2-15 years). Incomplete tumor resection with postoperative radiation therapy resulted in progressive tumor growth in three cases. One patient in this group experienced subarachnoid bleeding that had to be managed by salvage surgery. After primary radiation therapy, glomus jugulare tumors were still evident on magnetic resonance imaging scans, but showed no signs of disease progression. As a result of our experience, we found that a one-stage radical tumor resection performed in collaboration by otologic surgeons and neurosurgeons was the best treatment for patients with large glomus jugulare tumors. PMID- 10473830 TI - Neck dissection in the management of regional metastases in patients with undifferentiated nasopharyngeal carcinomas. AB - Residual regional disease after the primary treatment of nasopharyngeal carcinoma is still considered to be a therapeutic problem. The limitations of prophylactic radical radiation, further doses of irradiation as a useful salvage procedure, and the effects on vital structures were the reasons that we employed a therapeutic protocol consisting of radical neck dissection after 40 Gy of radiotherapy and a full tumor dose after surgery. The initial treatment consisted of chemotherapy. Between 1977 and 1991 surgical removal of residual neck metastases was performed in 44 patients with undifferentiated nasopharyngeal carcinomas who had regional metastases at the time of diagnosis. Fourteen patients (group A) had radical neck dissections after initial chemotherapy (using doxorubicin, etoposide, bleomycin and/or 5-fluouracil) and between two courses of locoregional radiotherapy. The remaining 30 patients (group B) were operated on after finishing chemotherapy and locoregional radiotherapy (group B 1) or receiving only full-dose locoregional radiotherapy (group B 2). All patients had histopathologically proven complete remission of primary tumors before neck surgery. The five-year survival rates for group A were 78%, 40% for group B 1 and 27% for group B 2. There were statistically significant differences between groups A and B (P < 0. 01), but not between groups B 1 and B 2. In group A one patient died from subsequent distant metastases and two from local tumor recurrences. Twenty patients died in group B, regional relapses occurred in 40% of the patients in group B 1 and 33% in group B 2, while distant metastases developed in 40% of group B 2. These findings again showed that radical neck dissection was an effective approach for controlling neck disease. When performed after initial chemotherapy and between two courses of radiotherapy, surgery significantly improves the prognosis of patients with positive regional lymph nodes at the time of diagnosis. PMID- 10473831 TI - Surgical management of acquired laryngopharyngeal fistulae. AB - Pathological communication between the food and air passages in the neck region due to malignant disease is known. However, such a pathology arising as a result of a non malignant process is relatively uncommon, and only a handful of reports exists in the literature. The authors describe and discuss the management of two patients with laryngopharyngeal fistula of nonmalignant etiology. PMID- 10473832 TI - Intramuscular hemangioma of the masseter muscle. AB - Intramuscular hemangiomas (IMH) are uncommon tumors of the head and neck, but often occur in the trunk and extremities. When present in the head, the masseter muscle is the most frequently involved site, although constituting only 0.8% of all hemangiomas. Accurate preoperative diagnosis is uncommon without cytology or biopsy. A case of IMH of the masseter muscle in a 24-year-old Turkish woman is presented. Clinical, radiologic and histologic findings and treatment modalities are reviewed. PMID- 10473833 TI - Recurrent respiratory papillomatosis associated with gastroesophageal reflux disease in children. AB - The hallmark of gastroesophageal reflux disease (GERD) is an increased exposure of esophageal and laryngeal mucosa to gastric juice. This exposure can cause complications such as chronic laryngitis or chronic respiratory diseases. We report our experience in managing three pediatric patients with severe recurrent juvenile laryngeal papillomatosis (JLP) associated with GERD. All patients showed a high rate of recurrence requiring multiple laser surgeries. Systemic alpha interferon therapy over a period of more than 1 year and photodynamic therapy with dihematoporphyrin produced no improvement. However, after therapy for GERD, the rate of recurrence of JLP decreased significantly. Although the course of respiratory papillomatosis is known to fluctuate, our findings suggest that gastroesophageal reflux may have a role in aggravating papillomatosis. PMID- 10473834 TI - Occupational health in Spain. PMID- 10473835 TI - An updated review of epidemiologic studies on the relationship between exposure to whole-body vibration and low back pain (1986-1997). AB - The aim of this study is to update the information on the epidemiologic evidence of the adverse health effects of whole-body vibration (WBV) on the spinal system by means of a review of the epidemiologic studies published between 1986 and 1997. In a systematic search, using several databases, of epidemiologic studies of low back pain (LBP) disorders and occupations with exposure to WBV, 45 articles were retrieved. The quality of each study was evaluated according to criteria concerning the assessment of vibration exposure, assessment of health effects, and methodology. The epidemiologic studies reaching an adequate score on each of the above-mentioned criteria were included in the final review. A meta analysis was also conducted in order to combine the results of independent epidemiologic studies. After applying the selection criteria, 17 articles reporting the occurrence of LBP disorders in 22 WBV-exposed occupational groups reached a sufficient score. The study design was cross-sectional for 13 occupational groups, longitudinal for four groups and of case-control type for one group. Two studies included both cross-sectional and follow-up data on the occurrence of LBP disorders in four occupational groups. The main reasons for the exclusion of studies were insufficient quantitative information on WBV exposure and the lack of control groups. The findings of the selected studies and the results of the meta-analysis of both cross-sectional and cohort studies showed that occupational exposure to WBV is associated with an increased risk for LBP, sciatic pain, and degenerative changes in the spinal system, including lumbar intervertebral disc disorders. Owing to the cross-sectional design of the majority of the reviewed studies, this epidemiologic evidence is not sufficient to outline a clear exposure-response relationship between WBV exposure and LBP disorders. Comparing the epidemiologic studies included in this review with those conducted before 1986, it is concluded that research design and the quality of exposure and health effect data in the field of WBV have improved in the last decade. PMID- 10473836 TI - Modeling of air pollution and its relationship with mortality and morbidity in Madrid, Spain. AB - OBJECTIVE: Evaluation of the association between air pollution and mortality and morbidity is becoming ever more complex owing to changes in inner-city air pollution, marked by decreasing values for all main pollutants save those associated with traffic. This has led to the need for the study of new epidemiological scenarios in which most pollutants are below guideline values. Nonetheless, the health effects are significant. METHODS: This report presents the results of a statistically based model for real-time forecasting of mortality and morbidity in Madrid, with meteorological and pollution series serving as inputs. RESULTS AND CONCLUSIONS: Not only did the models perform well with correlation coefficients between predicted and observed values (r = 0.683 for mortality, r = 0.681 for morbidity), but they enabled quantification of the impact of air pollution on mortality and morbidity (with increases ranging from 1. 8% to 12% for mortality and from 2.3% to 18% for morbidity for a 25 microg/m(3) increase in pollutants). Moreover, attention should be drawn to the observation that the model proved to be easy to implement and operate on a routine basis. PMID- 10473837 TI - Thermal thresholds, vibrotactile thresholds and finger systolic blood pressures in dockyard workers exposed to hand-transmitted vibration. AB - OBJECTIVES: To quantify neurological dysfunction in workers exposed to hand transmitted vibration using alternative neurological tests. To relate the neurological findings to the results of vascular tests and the symptoms reported by subjects with vibration-induced white finger. METHODS: Thermal thresholds (for perception of heat and cold), vibrotactile thresholds (for perception of vibration at 31.5 and 125 Hz) and finger systolic blood pressures were measured in 107 dockyard workers, including 31 controls and 76 workers exposed to hand transmitted vibration (50 reporting finger blanching consistent with vibration induced white finger). A history of vibration exposure and symptoms associated with hand-transmitted vibration were obtained for each subject. RESULTS: Increased duration of exposure to vibration resulted in a deterioration of both thermal thresholds and vibrotactile thresholds. Finger systolic blood pressures were lower in subjects reporting finger blanching and were related to the extent of blanching on the measured finger. Reported sensations of tingling were not correlated with any of the threshold measures; thermal thresholds and vibrotactile thresholds showed evidence of deterioration with reports of increasing numbness. Both numbness and tingling were correlated with reports of finger blanching. Finger systolic blood pressures were not correlated with either thermal or vibrotactile thresholds. CONCLUSIONS: Vascular and neurological signs produced by hand-transmitted vibration can occur independently, but the principal vascular symptom (i.e. attacks of blanching) and some commonly reported neurological symptoms (i.e. sensations of numbness and tingling) may be related. PMID- 10473838 TI - Surface EMG of shoulder and back muscles and posture analysis in secretaries typing at visual display units. AB - OBJECTIVE: A study was carried out to investigate temporal changes of activation of shoulder and back muscles in workers at visual display units by means of surface EMG. Moreover, postural parameters were recorded to distinguish fatigue related from posture-related changes of the myoelectrical activity. METHODS: Nine healthy female office workers typed texts spoken from tape during three 1-h-long sessions. After the first and again after the second hour there was a break of 15 min. Sixteen-channel surface EMG was bipolarly recorded from the erector spinae, trapezius, deltoid and sternocleidomastoid muscles. Root mean square (RMS) and power spectrum median frequency of the EMG were calculated. Sitting posture was assessed using an eight-channel movement analysis system with ultrasound markers. The position of the seventh cervical spinous process and the left and the right acromion were analysed synchronously with the EMG characteristics using regression analysis. RESULTS: The normalised RMS of the left and right trapezius muscle increased, while the median frequency did not change. The increase of the normalised RMS was significantly lower when the linear influence of posture was excluded. On average, the distance between C7 and the left and right acromion decreased within each working an hour. C7 became lower on average by 5.5 mm within an hour, whereas the acromions became lower by only 1.7 mm (left) and 3.3 mm (right). CONCLUSION: The increase in trapezius muscle activity was partly related to a lifting of the shoulders to compensate a slight slumping of the back. Another part of the EMG activity increase has to be attributed to fatigue, to attention-related activity or to the combination of both. Therefore, training of the back muscles and a varied organisation of work might have a preventive effect with respect to musculoskeletal complaints in VDU workers. PMID- 10473839 TI - Musculoskeletal disorders among female dental personnel--clinical examination and a 5-year follow-up study of symptoms. AB - OBJECTIVES: To explore the natural course of musculoskeletal disorders during a 5 year period among dental personnel, to survey findings and diagnoses according to a standardized protocol, and to compare and evaluate different methods for the detection of musculoskeletal disorders in a population. METHODS: In a 5-year follow-up study, dental personnel (n = 90) and referents (n = 30) were studied. The Nordic questionnaire (years 0 and 5), the present pain rating according to the Borg category ratio scale, and physical examination (year 5) were used. RESULTS: In year 0 the prevalence of symptoms in the shoulders, the wrists/hands, and, unexpectedly, the hips was higher in the dental personnel as compared with the referents. Furthermore, those (16%) who had left the dental profession during the observation period showed a higher prevalence of and, often, combined symptoms from several body regions in year 0 than did those who stayed. Dental personnel who remained in the profession tended to have an increased risk of developing more symptoms in the shoulders and the elbows/wrists/hands in year 5 as compared with year 0 and at year 5 were in more pain and had received more diagnoses for the neck/shoulder region relative to the referents. Furthermore, there was a considerable variation in symptoms during the follow-up period. The sensitivity of the pain rating and of the Nordic questionnaire in detecting musculoskeletal disorders was high for the neck and shoulders but was not as high for the elbow, wrists/hands, or hips. The association was better for diagnoses than for findings. The opposite patterns were observed for specificity. CONCLUSIONS: Dental personnel had an increased risk of developing musculoskeletal disorders as verified by symptoms and diagnoses and more painful or persistent conditions. This led to a selection out of work. The questionnaire and the present pain rating gave a relatively good picture of the prevalence of musculoskeletal disorders arising from the neck, shoulders, and hips and would be useful as screening tools. Their sensitivity in detecting disorders was higher for diagnoses than for findings. However, these methods were not as sensitive for disorders involving the elbows/wrists/hands. Physical examinations gave more detailed information. PMID- 10473840 TI - The balloon technique: a convenient method to measure exhaled NO in epidemiological studies. AB - OBJECTIVES: The aim of the present study was to evaluate the balloon procedure to measure exhaled nitric oxide (NO). METHODS: This was performed by comparing the procedure with the well-established on-line measurement of NO (direct exhalation into the NO module). Using both procedures exhaled NO was measured in 16 healthy subjects on two days with different level of air pollution. RESULTS: Exhaled NO measured on-line was 3.8-4.5 times lower than exhaled NO obtained using the balloon technique but the two sets of values correlated linearly (r 0.93-0.97). Mean NO level on day 1 with low air pollution and day 2 with high air pollution was 6.6 and 8.1 parts per billion (ppb; on-line measurement) and 25.2 and 36.9 ppb (balloon method), respectively. The day 1 to day 2 ratio differed per subject but was independent of the technique of measurement. Mean day-to-day ratio of exhaled NO using the balloon technique (1. 65 +/- 0.13) was not different (P < 0.05) from the ratio of NO levels measured on-line (1.49 +/- 0.13). Based on these ratios the increase in level of outdoor air pollution appears to be associated with a 49-65% increase in exhaled NO. CONCLUSION: Exhaled NO is proposed as a de novo individual biomarker to monitor the adverse effects of air pollution. The balloon procedure offers a sound and convenient alternative for the on-line procedure to measure exhaled NO in large populations as required in epidemiological studies. PMID- 10473841 TI - Acute symptoms during non-inhalation exposure to combinations of toluene, trichloroethylene, and n-hexane. AB - OBJECTIVES: To study the acute effect of exposure to a mixture of three commonly used solvents in humans using a route of exposure not involving the nose and lungs, in this case a gastrointestinal application. METHODS: In a 2(3)-factorial experiment eight healthy male volunteers were exposed to the eight combinations of toluene (1. 5 and 4 mg. min(-1)) trichloroethylene (1.5 and 4 mg. min(-1)), and n-hexane (0.3 and 1.0 mg. min(-1)) for 60 min given into the stomach via a feeding tube. The body burden was measured by the exhaled solvent concentrations and the urinary excretion of metabolites during and after exposure. The subjective ratings of tiredness, sleepiness, headache, nausea, feeling of intoxication, and dizziness were estimated by continuous linear analogue rating scales before and 30, 90, and 240 min after the start of exposure. RESULTS: Concentrations of the three solvents in the end exhaled air varied between 0 and 46 mg. min(-3). The ratings of symptoms were generally low and there was no difference between the high and low doses. Neither was there any correlation between the concentration of solvents in the exhaled air and the ratings. CONCLUSIONS: The study show no effects at levels in end exhaled air of mixtures of solvents which in inhalation studies have given signs of a possible neurotoxic effect. The lack of symptoms using this alternative route supports the hypothesis that "neurotoxic" symptoms in relation to exposure to very low air concentrations of solvents are mainly indirect, mediated by irritation or smell. PMID- 10473842 TI - Remodeling of the occupational medical examination program in South Korea. AB - The South Korean government carried out reforms in the occupational medical examination program over a period of 2 years from 1997 to 1998 in a response to the discontent of workers, who had complained that occupational medical examinations were merely ritualistic and unproductive. The purposes of this manuscript are to describe the reason for the remodeling, with a historical review of the existing occupational medical examination program in South Korea, and to discuss the main issues involved in remodeling the program. The existing occupational medical examination program in South Korea was reviewed and criticized. The basic philosophy and the main issues in remodeling South Korea's occupational medical examination program were also discussed. In conclusion, the occupational medical examination program should not be conducted in a uniform manner according to legal regulations, but should be a part of overall occupational health services. Individual workplaces should be given the autonomy to perform medical examinations according to the characteristics of the workplace. An enabling approach based on the participation of labor and management should be taken into consideration. PMID- 10473843 TI - Identification of head motions by central vestibular neurons receiving linear and angular input. AB - Most naturally occurring displacements of the head in space, due to either an external perturbation of the body or a self-generated, volitional head movement, apply both linear and angular forces to the head. The vestibular system detects linear and angular accelerations of the head separately, but the succeeding control of gaze and posture often relies upon the combined processing of linear and angular motion information. Thus, the output of a secondary neuron may reflect the linear, the angular, or both components of the head motion. Although the vestibular system is typically studied in terms of separate responses to linear and angular acceleration of the head, many secondary and higher-order neurons in the vestibular system do, in fact, receive information from both sets of motion sensors. The present paper develops methods to analyze responses of neurons that receive both types of information, and focuses on responses to sinusoidal motions composed of a linear and an angular component. We show that each neuron has a preferred motion, but a single neuron cannot code for a single motion. However, a pair of neurons can code for a motion by the relative phases of firing-rate modulation. In this way, information about motion is enhanced by neurons combining information about linear and angular motion. PMID- 10473844 TI - Neuromolecular computing: a new approach to human brain evolution. AB - Evolutionary approaches in human cognitive neurobiology traditionally emphasize macroscopic structures. It may soon be possible to supplement these studies with models of human information-processing of the molecular level. Thin-film, simulation, fluorescence microscopy, and high-resolution X-ray crystallographic studies provide evidence for transiently organized neural membrane molecular systems with possible computational properties. This review article examines evidence for hydrophobic-mismatch molecular interactions within phospholipid microdomains of a neural membrane bilayer. It is proposed that these interactions are a massively parallel algorithm which can rapidly compute near-optimal solutions to complex cognitive and physiological problems. Coupling of microdomain activity to permenant ion movements at ligand-gated and voltage-gated channels permits the conversion of molecular computations into neuron frequency codes. Evidence for microdomain transport of proteins to specific locations within the bilayer suggests that neuromolecular computation may be under some genetic control and thus modifiable by natural selection. A possible experimental approach for examining evolutionary changes in neuromolecular computation is briefly discussed. PMID- 10473845 TI - On the directionality of cortical interactions studied by structural analysis of electrophysiological recordings. AB - To investigate the directionality of neural interactions as assessed by electrophysiology, we adapted methods of structural analysis from the field of econometrics. In particular, within the framework of autoregressive modelling of the data, we considered quantitative measures of linear relationship between multiple time series adopting the Wiener-Granger concept of causality. The techniques were evaluated with local field potential measurements from the cat visual system. Here, several issues had to be addressed. First, out of several statistical tests of the stationarity of local field potentials considered, those based on the Kolmogorov-Smirnov and on the reverse arrangement statistics proved to be most powerful. The application of those tests to the experimental data showed that the large part of the local field potentials can be considered stationary on a time scale of 1 s. Second, out of the several investigated methods for the determination of an optimal order of the autoregressive model, the Akaike Information Criterion had the most suitable properties. The identified order of the model, across different repetitions of the trials, was consistently 5-8. Third, although the individual segments of field potentials used for the analysis were relatively short, the methods of structural analysis applied produced reliable results, confirming findings of simulations of data with similar properties. Furthermore the features of the estimated models were consistent among trials, so that the analysis of average measures of interaction appears to be a viable approach to investigate the relationship between the recording sites. In summary, the statistical methods considered have proved to be suitable for the study of the directionality of neuronal interactions. PMID- 10473847 TI - Passive bipedal walking with phasic muscle contraction. AB - The existence of self-organizing walking patterns is often considered the result of a mechanical system interacting with the environment and a (neural) oscillating unit. The pattern generators might be thought of as an indispensable component for the existence of limit cycle behavior. This paper shows that this is not a necessity for the existence of a self-organizing bipedal walking pattern. Stable walking cycles emerge from a simple passive bipedal structure, with an energy source inevitably present to sustain the oscillation. In this work the energy source is chosen to be phasic muscle contraction. A two-dimensional model is composed of two legs and a hip mass, symbolizing the trunk. The stance leg stiffness is generated by two muscles. The hip stiffness is generated by four muscles. Muscle activation is caused by two reflex-like trigger signals, without feedback control. Human equivalent model parameters such as geometry and mass distribution were assumed. With return map analysis, the model is analyzed on periodic behavior. Stable walking cycles were found and could be manipulated during walking by varying the muscle or reflex parameters, forcing the oscillation to converge to a new attractor. PMID- 10473846 TI - Emergence of symmetric, modular, and reciprocal connections in recurrent networks with Hebbian learning. AB - While learning and development are well characterized in feedforward networks, these features are more difficult to analyze in recurrent networks due to the increased complexity of dual dynamics - the rapid dynamics arising from activation states and the slow dynamics arising from learning or developmental plasticity. We present analytical and numerical results that consider dual dynamics in a recurrent network undergoing Hebbian learning with either constant weight decay or weight normalization. Starting from initially random connections, the recurrent network develops symmetric or near-symmetric connections through Hebbian learning. Reciprocity and modularity arise naturally through correlations in the activation states. Additionally, weight normalization may be better than constant weight decay for the development of multiple attractor states that allow a diverse representation of the inputs. These results suggest a natural mechanism by which synaptic plasticity in recurrent networks such as cortical and brainstem premotor circuits could enhance neural computation and the generation of motor programs. PMID- 10473848 TI - The role of neuronal death during the development of topographically ordered projections: a computational approach. AB - At the time of synaptogenesis typically 50% of the neurons die. The biological role of this is still unclear, but there is evidence in the visual system that many neurons projecting to topographically inappropriate parts of their target are eliminated to improve the accuracy of the mapping. The signaling that determines neuronal survival involves electrical activity and trophic factors. Based on these observations, we have elaborated a computational model for the self-organization of a two-layered neural network. We observe changes in the topographical organization between the two layers. In layer 1, a traveling wave of electrical activity is used as input. Activity transmission to layer 2 can generate, according to a Hebbian rule, a retrograde death signal that is compensated by a trophic survival signal generated by the target cells. Approximately 50% of the neurons die, and we observe refinement in the topography between the two layers. In alternative versions of the model, we show that an equivalent reorganization can occur through Hebbian synaptic modification alone, but with less precision and efficiency. When the two mechanisms are combined, synaptic modification provides no further improvement over that produced by neuronal death alone. This computational study supports the hypothesis that neuronal death during development can play a role in the refinement of topographical projections in the nervous system. PMID- 10473849 TI - A comparison of models explaining muscle activation patterns for isometric contractions. AB - One of the main problems in motor-control research is the muscle load sharing problem, which originates from the fact that the number of muscles spanning a joint exceeds the number of degrees of freedom of the joint. As a consequence, many different possibilities exist for the activation of muscles in order to produce a desired joint torque. Several models describing muscle activation have been hypothesized over the last few decades to solve this problem. This study presents theoretical analyses of the various models and compares the predictions of these models with new data on muscle activation patterns for isometric contractions in various directions. None of the existing models fitted the experimental data in all aspects. The best fit was obtained by models based on minimization of the squared sum of muscle forces ( summation operator(m)φ(2)(m), which is almost equivalent to the Moore-Penrose pseudo inverse solution), muscle stress sigma ( summation operator(m)sigma(m)(2)) or muscle activation alpha ( summation operator(m)alpha(m)(2)). Since muscle activation patterns are different for isometric contractions and for movements, it could well be that other models or optimization criteria are better suited to describe muscle activation patterns for movements. The results of our simulations demonstrate that the predicted muscle activation patterns do not depend critically on the parameters in the model. This may explain why muscle activation patterns are highly stereotyped for all subjects irrespective of differences between subjects in many neuro-anatomical aspects, such as, for example, in the physiological cross-sectional area of muscle. PMID- 10473850 TI - Evidence for different nonlinear summation schemes for lines and gratings at threshold. AB - Within a wide class of multichannel models of the visual system it is suggested that spatial distributions of luminance are processed by the independent activation of grating detectors, or spatial frequency channels. Probability summation is often described in terms of Quick's nonlinear pooling model [Quick RF (1974) Kybernetik 16:65-67]. Using this model, we find evidence for the existence of different kinds of nonlinear summation at threshold; for compound gratings with well-separated spatial frequency components, the threshold functions indicate nonlinear summation which is not compatible with probability summation, while for line patterns well separated in the spatial domain the probability summation rule proves compatible with the data. PMID- 10473851 TI - Resection of colorectal liver metastases. AB - INTRODUCTION: Surgical resection is presently the only approach that offers patients with liver metastases from colorectal carcinoma substantial chance of cure. This article summarizes the current literature as well as the author's personal experience. BACKGROUND AND DISCUSSION: Since 1980, 5-year survival figures have ranged from 21% in collected series to 48% in single-institution series. The 30-day mortality of elective liver resection in non-cirrhotic patients ranges now between 0% and 5%. The overwhelming indicator of prognosis is the completeness of tumor removal according to the R-classification. The specific impact of all other factors should therefore be analyzed by excluding non-radical procedures and operative mortality. Among patient characteristics, age and gender do not significantly affect outcome, while the Karnofski stage is important. Regarding the primary tumor, the effect of staging and location is predominantly apparent in patients with synchronous metastases. Timing of metastasis detection is of some importance, as most authors found a slightly better outcome for metachronously detected metastases. With respect to the liver involvement, multiplicity of metastases and bilateral disease both seem to be of minor importance after R0-resection, while satellite lesions are significant in many series. The actual number of metastases is of minor effect, with a slight superiority in 5-year survival for patients with one to three nodules relative to patients with four nodules or more in most series, but identical results in the author's own experience. The maximum diameter as an indicator of tumor burden represents a significant prognosticator in half of the reports analyzed. Extrahepatic disease reduces 5-year survival, but direct tumor invasion to adjacent structures, local recurrent disease, or one or few pulmonary metastases are no contraindication to liver resection as long as a R0-situation can be achieved. In contrast, lymph-node metastases at the liver hilum predict a poor outcome. They are likely to prove as a clear contraindication. With respect to the operative approach, a clear margin of 1 cm or more should be aimed at but, if the size or location of metastases do not allow a 1-cm margin, resection should still be performed, making every surgical effort to ensure a complete rim of unaffected tissue. Anatomic resections reduce the incidence of non-radical procedures and may improve survival. Whether there is an independent effect of operative blood loss, need for blood transfusion, and intraoperative hypotension on prognosis is still unclear. Adjuvant chemotherapy or radiotherapy after R0 resection is unlikely to improve results. There are also no convincing data available demonstrating a prognostic benefit when a non-curative resection is supplemented by any medical treatment. In patients with recurrent disease, a re resection is possible in roughly 20%. Survival from the time of re-intervention ranges from 21% to 57% after 5 years and, thus, justifies a close follow-up policy after R0-resection of the initial liver metastases. CONCLUSION: The previous "clear" contraindications to liver resection have become less important. Future efforts may be directed to more accurate patient selection and new approaches of neoadjuvant and adjuvant therapeutic strategies. PMID- 10473852 TI - Relevance of neoadjuvant and adjuvant treatment for patients with resectable liver metastases of colorectal carcinoma. AB - BACKGROUND: Excellent results after resection of colorectal liver metastases are associated with a high rate of recurrence. Influenced by positive results of palliative and adjuvant treatment in advanced cancer, various chemotherapy regimens were evaluated to improve long-term results. METHODS: The databases Medline and Cancerlit (1982-1998) gave information about 675 patients who were treated either by means of systemic, intra-arterial, intraportal or intraperitoneal administration before or after liver resection. RESULTS: In general, the feasibility of an adjuvant treatment was tested. Proof has been furnished for the practicability of systemic and arterial therapy and for immunotherapy after liver resection whereas, for peritoneal and portal treatment, further studies are necessary. In a few non-randomised trials, it has been possible to discern a trend towards an improvement due to adjuvant postoperative therapy using historical or matched-pair control groups. Until now, only one of five randomised studies has been published. Six months of postoperative adjuvant intra-arterial treatment using 5-fluorouracil (1000 mg/m(2) for 5 days every 28 days) and folinic acid (200 mg/m(2) for 5 days every 28 days) was compared with observation only. Neither in the intention-to-treat nor in the as-treated analysis was median survival time (34.5 months versus 40. 8 months and 39.7 months versus 44.8 months, respectively) significantly increased. As neoadjuvant treatment was successful in primary non-resectable patients, this approach is now being tested in resectable patients. CONCLUSION: Despite several theoretical reasons for post- or preoperative treatment in resectable patients, every approach should be tested using of controlled studies. PMID- 10473853 TI - Percutaneous ethanol injection and radio-frequency ablation for the treatment of nonresectable colorectal liver metastases - techniques and results. AB - BACKGROUND: Percutaneous ethanol injection (PEI) and radio-frequency (RF) ablation are possible palliative treatment modalities for patients with non resectable liver metastases of colorectal carcinomas. The different techniques are explained and reviewed. RESULTS: PEI did not show promising results for the treatment of liver metastases. RF results were more encouraging; some studies showed improved mean survival times for patients when a complete necrosis of the metastases could be achieved. The maximum diameter of the necrotic area possible in a single session is about 5 cm. CONCLUSION: PEI and RF are palliative last line treatment strategies for patients with non-resectable liver metastases and should only be applied if chemotherapy is not sufficient or not possible. The long-term efficacy of RF ablation in this group of patients has to be evaluated. PMID- 10473854 TI - Regional chemotherapy of non-resectable liver metastases from colorectal cancer - literature and institutional review. AB - BACKGROUND: Cure is possible by resecting colorectal isolated liver metastases. In non-resectable isolated colorectal liver metastases (CRLM), regional chemotherapy has been advocated to optimize the disease control in the liver in order to improve the results of the alternative, systemic chemotherapy. The drugs are delivered by means of hepatic artery infusion (HAI) via ports or pumps; pharmacological modifications of the hepatic arterial blood-flow-like HAI with starch microspheres or stop-flow and perfusion techniques were applied to improve HAI. METHODS: We reviewed the literature and report our progress, up to May 1999, in analyzing the validity of HAI for CRLM therapy. RESULTS: In the majority of phase-II and -III trials, the response rates to HAI were significantly higher than those from systemic chemotherapy, and local disease control could be achieved even when HAI was used second line to systemic chemotherapy. The meta analysis of randomized trials comparing HAI with either systemic chemotherapy (five trials) or, optionally, either 5-fluorouracil (FU) or symptomatic treatment (two trials) showed a significant advantage of HAI in response (41% vs 14%, P<10( 10)) and median survival time (15 months vs 11 months, P<0.0009). The active anabolite of 5-FU, 5-fluordeoxyuridine (5-FUDR), the drug of choice for HAI in those trials, may cause severe hepatotoxicity. To avoid this toxicity, we developed a HAI protocol using mitoxantrone, 5-FU plus folinic acid (FA) and mitomycin C (MFFM). The response rates of HAI with 5-FU plus FA or MFFM were 45% and 66%, the interim median survival times 19.8 months and 27.4 months. 5-Year survivors were observed in all our protocols. Since no severe hepatotoxicity occurred, 9 of 74 patients were resected after response to HAI with 5-FU plus FA or MFFM, without surgical mortality and with survival times from 2+ months to 58+ months. CONCLUSION: The high response rates, the long survival times, the possibility of achieving 5-year-survival either by HAI alone or by resection after down staging with HAI all sum up to the evidence that HAI could be the primary choice of treatment for CRLM. Phase-III trials are conducted to compare the protocols with optimal regional versus systemic chemotherapy. PMID- 10473855 TI - Is delayed gastric emptying following pancreaticoduodenectomy related to pylorus preservation? AB - BACKGROUND: Delayed gastric emptying (DGE) is the most frequent postoperative complication after pylorus-preserving pancreaticoduodenectomy (PPPD). This prospective, non-randomized study was undertaken to determine whether the incidence of DGE may be reduced by modifying the original reconstructive anatomy with a retrocolic duodenojejunostomy towards an antecolic duodenojejunostomy. PATIENTS AND METHODS: The study was comprised of 51 patients who underwent PPPD between August 1994 and November 1997. The operation was carried out as originally described but was modified by performing the duodenojejunostomy antecolically. Clinical data were recorded prospectively, with special regard to DGE. RESULTS: After PPPD, the nasogastric tube could be removed at a median of 2 days (range 1-22 days) postoperatively; in two patients, the nasogastric tube was reinserted because of vomiting and nausea. A liquid diet was started at a median of 5 days (3-11 days); the patients were able to tolerate a full, regular diet at a median of 10 days (7-28 days). The overall incidence of DGE was 12% (n=6). No postoperative complications other than DGE were exhibited by 36 patients (71%). In this group, DGE was only seen in one patient (3%). In the second group, where postoperative complications other than DGE occurred (n=15), five patients (30%) exhibited DGE (P=0.002). CONCLUSIONS: DGE after PPPD seems to be of minor clinical importance following uncomplicated surgery. When taking the results into consideration, it can be said that, despite the lack of a control group, antecolic duodenojejunostomy might be the key to a low incidence of DGE after PPPD. In our experience, DGE is linked to the occurrence of other postoperative complications rather than to pylorus preservation. PMID- 10473856 TI - Laparoscopic ultrasonography reliably visualizes bile-duct injuries - a blind randomized porcine study. AB - INTRODUCTION: Despite use of intraoperative cholangiography (IOC), only one-third or less of bile duct injuries that occur during laparoscopic cholecystectomy are detected at the primary operation. Therefore, there is a need to investigate other intraoperative diagnostic methods, especially laparoscopic ultrasound, for detecting bile-duct injuries. MATERIALS AND METHODS: Detection of different bile duct injuries by laparoscopic intraoperative ultrasonography was evaluated in a blind, randomized animal study. Fifty bile-duct injuries were created using laparoscopic techniques in 23 pigs. The number of single lesions and the order in which the injuries were created was randomly assigned. The lesions created were: (1) partial occlusion by a clip, (2) complete occlusion by a clip, (3) partial laceration, (4) transection without clips, (5) transection between clips and (6) excision between clips. In addition, there were cases without lesions. Different types of clips were used. A screening of the injuries by laparoscopic ultrasound using a flexible probe was attempted. The study was performed in a blind fashion. RESULTS: Complete ultrasonographical imaging of the hepatic and common bile duct between its bifurcation and its insertion into the duodenum was accomplished in all 23 animals before surgery was performed. Eight cases without lesions were correctly recognized. In 48 of 49 cases, the injuries were detected (sensitivity 98%). In one instance, the injury was obvious without the use of ultrasound. Forty-four cases were correctly diagnosed and, in 12 cases, the injuries were equivocally differentiated (specificity of 100%, overall accuracy of 98.3%). There was only one false diagnosis. The different types of clips were also reliably differentiated. CONCLUSIONS: We conclude that laparoscopic ultrasonography can reliably visualize most relevant bile-duct injuries. A standard for doing the ultrasound examination is proposed. Direct and indirect ultrasound signs of bile-duct injuries are explained. PMID- 10473857 TI - Should laparoscopic approach be proposed for large and/or potentially malignant adrenal tumors? AB - INTRODUCTION: Laparoscopic adrenalectomy (LA) is safe and effective for small, benign, functioning tumors. Whether it should be performed for other adrenal tumors is questionable. The aim of this study was to evaluate and compare the complications and results of 150 consecutive LAs performed either for small benign tumors or for large and/or potentially malignant tumors. METHODS: Between June 1994 and August 1998, we performed 150 LAs in 142 patients. We used a transperitoneal flank approach in the lateral decubitus position. Initially, our indications for LA were limited to small (<4 cm) benign tumors (group I, n=102): 56 aldosteronomas, 33 Cushing's syndrome, 11 pheochromocytomas and 2 nonfunctional tumors. Progressively, based on increasing experience, LA was also proposed for tumors larger than 4 cm or potentially malignant (group II, n=48): 5 Cushing's syndrome, 1 androgen-producing tumor, 14 pheochromocytomas and 28 nonfunctional tumors. Preoperative demonstration of invasive extra-adrenal carcinoma remained an absolute contraindication for LA. RESULTS: Mean tumor size was 21.1 mm in group I and 51.6 mm in group II. All tumors in group I were benign. Six of the 48 tumors in group II were malignant (12.5%). The rate of complication was, respectively, 7.8% and 8.3% in groups I and II. The rate of conversion was, respectively, 4.9% and 6.2% in groups I and II. Mean operative time was 131 min in group I and 129 min in group II. The endocrinopathy was cured in all patients. To date, no recurrences have been observed. CONCLUSIONS: LA can be proposed for large (<12 cm) or potentially malignant adrenal tumors provided preoperative investigations have not demonstrated invasive carcinoma. An open procedure should be performed instead if local invasion is observed at the start of the operation. PMID- 10473858 TI - Total hepatectomy and liver transplantation for metastatic neuroendocrine tumors of the pancreas - a single center experience with ten patients. AB - BACKGROUND: Metastatic neuroendocrine pancreatic tumors have a poor prognosis. We have studied retrospectively the efficacy of liver transplantation as ultimate therapy of otherwise untreatable symptomatic neuroendocrine hepatic metastases originating in the pancreas. METHODS: We reviewed our experience of liver transplantation (LTx) for hepatic metastases of neuroendocrine pancreatic tumors in ten patients. The indication for liver grafting was seen in cases of irresectable metastases and when patients were suffering from otherwise untreatable tumor-associated symptoms due to massive hormonal release or large intra-abdominal tumor bulk. RESULTS: In four patients, the primary tumors had been removed before LTx, in five patients simultaneously with LTx and in one case 46 months after grafting. There was no operative mortality. After hepatectomy and LTx, all patients had complete relief of symptoms and all preoperatively increased hormonal levels returned to normal. In nine of ten patients, the transplant procedure had the potential for cure, whereas, in one patient, the primary tumor had remained in situ at LTx and was removed 46 months later by an R2-resection. At present, nine patients are alive with a median follow-up of 33 months (range 13.5 months to 117 months). The one patient in whom the primary tumor was removed after transplantation died due to massive intra-abdominal tumor spread 68 months after LTx. Currently, two patients are without evidence of disease, but one of them after re-operation because of lymph-node metastases 8 months after transplantation. The longest disease-free survival is now more than 7 years. In seven of nine patients, tumor recurred between 1.5 months and 48 months after transplantation. CONCLUSIONS: Patients with otherwise untreatable symptomatic neuroendocrine hepatic metastases of pancreatic origin may benefit from total hepatectomy and liver transplantation with regard to symptomatic relief and long-term survival, despite frequent recurrence of disease. In some patients, liver transplantation may even offer the chance for cure. PMID- 10473859 TI - Variations of surgical reconstruction in liver transplantation depending on vasculature. AB - BACKGROUND: From September 1988 through April 1998, 1000 liver transplantations were performed on 911 patients. The standard technique for liver re vascularization to guarantee an optimal blood inflow during transplantation was modified in 19% of the cases on the arterial side and in 5.6% of the cases on the portal side as a result of unusual anatomical features and pathological changes in the vasculature of the organ recipient. In 113 transplantations, successful reconstruction of accessory vessels of the graft (12 left and 101 right hepatic arteries) was performed without complications. It is our opinion that preoperative diagnosis of the vasculature (stenoses of the celiac trunk etc.) of the organ recipient by duplexsonography and angiography is necessary. Even with the help of these tests, it is extremely difficult to diagnose a "steal" syndrome in the splenic artery: for example, 31 of 40 patients with poor liver function received postoperative therapy for newly diagnosed "steals". RESULTS AND CONCLUSIONS: There is no increase in complications (stenosis and thrombosis) with modifications of arterial reconstruction (4.9 vs 6.3%); however, with modification of portal reconstruction the increase is from 2.4% to 8.3%. PMID- 10473860 TI - Attitude of patients toward transplantation of xenogeneic organs. AB - BACKGROUND: The prospect of xenotransplantation has stimulated considerable hopes as well as major concerns. The question of whether or not patients accept xenografts is influenced not only by scientific facts but also by psychological factors. It was the aim of this study to analyze the attitudes of patients toward transplantation of xenogeneic organs and evaluate factors influencing these attitudes. METHODS: To this end, attitudes toward xenogeneic compared with allogeneic organ grafts were evaluated by means of detailed questionnaires in 1049 patients in Germany, who either had received transplants (n=722) or were on the waiting list for various organ grafts (n=327). Answers were correlated to demographic data as well as to the physical and mental conditions of the patients. RESULTS: The survey indicates that 77% of patients would accept xenografts while 7% would refuse them if results of xenotransplantation were comparable with those of allotransplantation. If xenotransplantation were associated with increased risks due to more intensive medication 58% would still basically accept xenografts. Acceptance of xenografts was significantly higher in patients who had received transplants and among males. Age, religion, waiting time, and type of organ were not found to influence acceptance rates. Xenografts were thought to be associated with considerable or severe emotional stress by 23% of patients, versus 3% for allografts. The pig was the preferred donor animal, and gene therapeutic manipulation for improvement of results would be accepted by 84%. Inadequate graft function/increased risk of rejection and risk of disease transmission were the major concerns for 60% and 52% of patients, respectively; emotional concerns were the major concerns for 24% and animal-rights concerns for 15%. CONCLUSIONS: These findings show that the potential acceptance rate of xenografts would be quite high, with a more positive attitude in transplanted patients than in waiting-list patients; there was no major difference in acceptance rate for various types of organs. Major concerns about xenotransplantation currently are functional inferiority and transmission of diseases. PMID- 10473861 TI - Intraoperative electromyogram monitoring of the recurrent laryngeal nerve: experience with an intralaryngeal surface electrode. A method to reduce the risk of recurrent laryngeal nerve injury during thyroid surgery. AB - INTRODUCTION: A clinical method to localize the recurrent laryngeal nerve intraoperatively in order to minimize the risk of accidental injury is presented. METHODS AND RESULTS: By means of an electrode, the nerve was stimulated by a pulsed contact current. The resulting muscle potential was detected using an electrode placed in the larynx. We applied this technique during the time period between 1 January 1997 and 31 December 1998. In 96 cases of primary thyroid resection, the recurrent nerve was identified 167 times. The patients were operated on for nodular thyroid (n=85), Grave's disease (n=9) and malignant papillary goiter (n=2). Retrospectively, the rate of intraoperative nerve injury, equivalent to the rate of postoperative transient and permanent nerve palsy, was 1.04% in the 96 patients and 0.60% with respect to the 167 nerves at risk. The rate of failure of the method was 7.29%. PMID- 10473862 TI - Immunotherapy in pancreatic cancer - current status and future. AB - BACKGROUND: In pancreatic cancer, multimodal protocols, involving chemotherapy, radiation, or regional treatment, are initiated to improve oncological outcome. Since pancreatic adenocarcinoma has been shown to be susceptible to immune stimulation, several immunotherapy approaches have been investigated to define the role of immunotherapy in pancreatic cancer. METHOD: A review of current and past data concerning experimental and clinical immunotherapy in pancreatic cancer is presented in the context of basic immunotherapeutic principles. Past pitfalls and future developments are analyzed and a synthesis of immune stimulation and immune suppression is deduced on the basis of published data. RESULTS: Preclinical and initial clinical studies with monoclonal antibodies CO17-1A, BW494/32 and anti-epidermal growth factor receptor (EGFR) have been conducted, and various targets suitable for immunotherapy have been identified involving new molecular and gene technology. Targets on pancreatic cancer cells currently under investigation are mucins (MUC-1), glycoproteins (GA733), ras peptides and EGFRs. Side effects are minor and rarely auto-immune reactive. Another approach combines randomized regional with systemic chemoimmunotherapy (mitomycin C, 5 fluorouracil, folinic acid, carboplatin, epirubicin; interferon-gamma, interleukin-2) in nonresectable pancreatic cancer and obtains significant differences in median survival rates (14 months vs 4.5 months in controls) and quality of life. CONCLUSION: Although single remarkable improvements in the immunological approach to treatment of pancreatic cancer have been made, immunotherapy in pancreatic cancer is still experimental. On the basis of reliable preclinical data, new immunotherapy protocols will have to be evaluated clinically. Careful monitoring of immune responses and side effects, and assessment of quality of life will ensure identification of effective immunotherapy protocols for human pancreatic cancer in the near future. PMID- 10473863 TI - Radical resection with autonomic nerve preservation and lymph node dissection techniques in lower rectal cancer surgery. PMID- 10473864 TI - Reply. PMID- 10473865 TI - neu mutation in schwannomas induced transplacentally in Syrian golden hamsters by N-nitrosoethylurea: high incidence but low allelic representation. AB - Peripheral nerve tumors (PNT) and melanomas induced transplacentally on day 14 of gestation in Syrian golden hamsters by N-nitrosoethylurea were analyzed for activated oncogenes by the NIH 3T3 transfection assay, and for mutations in the neu oncogene by direct sequencing, allele-specific oligonucleotide hybridization, MnlI restriction-fragment-length polymorphism, single-strand conformation polymorphism, and mismatch amplification mutation assays. All (67/67) of the PNT, but none of the melanomas, contained a somatic missense T --> A transversion within the neu oncogene transmembrane domain at a site corresponding to that which also occurs in rat schwannomas transplacentally induced by N nitrosoethylurea. In only 2 of the 67 individual hamster PNT did the majority of tumor cells appear to carry the mutant neu allele, in contrast to comparable rat schwannomas in which it overwhelmingly predominates. The low fraction of hamster tumor cells carrying the mutation was stable through multiple transplantation passages. In the hamster, as in the rat, specific point-mutational activation of the neu oncogene thus constitutes the major pathway for induction of PNT by transplacental exposure to an alkylating agent, but the low allelic representation of mutant neu in hamster PNT suggests a significant difference in mechanism by which the mutant oncogene acts in this species. PMID- 10473866 TI - Expression of membrane type 1 matrix metalloproteinase, matrix metalloproteinase 2 and tissue inhibitor of metalloproteinase 2 in human cartilaginous tumors with special emphasis on mesenchymal and dedifferentiated chondrosarcoma. AB - Membrane type 1 matrix metalloproteinase (MT1-MMP) has been identified as an activator of the proenzyme of matrix metalloproteinase 2 (MMP-2: gelatinase A), and has also been shown to play a crucial role in tumor invasion by activating proMMP2 in both lung and gastric carcinoma. The tissue inhibitor of metalloproteinase 2 (TIMP-2) plus the MT1-MMP complex also plays an important role in the activation of proMMP-2. In this study, the expressions of MT1-MMP, MMP-2 and TIMP-2 were evaluated in 10 enchondromas, 34 conventional chondrosarcomas, 5 clear-cell chondrosarcomas, 7 mesenchymal chondrosarcomas and 8 dedifferentiated chondrosarcomas. The expressions were immunohistochemically visualized on paraffin sections and the levels of expression were assessed semiquantitatively. The extent of staining was assessed by the extent score in order to determine the overall level of expression. The extent scores of MT1-MMP, MMP-2 and TIMP-2 in grade 2 chondrosarcoma were significantly higher than those in either enchondroma or grade 1 chondrosarcoma (P < 0.05). In conventional chondrosarcoma, significant correlations were found between the extent scores of MT1-MMP and MMP-2 (P < 0.001), MT1-MMP and TIMP-2 (P < 0.01), and MMP-2 and TIMP 2 (P < 0.01). The undifferentiated small round tumor cells of mesenchymal chondrosarcoma showed lower positive rates and extent scores for MT1-MMP (2/7, 0.7 +/- 0.5) and MMP-2 (3/7, 0.7 +/- 0.4) than for cartilaginous components of mesenchymal chondrosarcoma [MT1-MMP (4/7, 1.3 +/- 0.5) and MMP-2 (7/7, 1.9 +/- 0.3)] or conventional chondrosarcoma. In dedifferentiated chondrosarcoma, the extent scores of MT1-MMP, MMP-2 and TIMP-2 in low-grade cartilaginous components were not significantly different from those in conventional chondrosarcoma; however, the high-grade anaplastic components showed high extent scores for MT1 MMP, MMP-2 and TIMP-2, compared with the low-grade cartilaginous components of dedifferentiated chondrosarcoma or conventional chondrosarcoma. According to our results, the expression of MT1-MMP as well as that of MMP-2 or TIMP-2 demonstrated a significant correlation with the tumor grade in human cartilaginous tumors. Furthermore, the expressions of MT1-MMP, MMP-2 and TIMP-2 were also found to play a crucial role in invasion in the high-grade components of dedifferentiated chondrosarcoma. PMID- 10473867 TI - Inactivation of p53 and of pRb protects human colorectal carcinoma cells against hyperthermia-induced cytotoxicity and apoptosis. AB - Cell-cycle checkpoints are thought to govern the cellular response to external stimuli. The involvement of the p53 tumour-suppressor protein and the retinoblastoma protein (pRb) in the cell-cycle checkpoint in G1 phase is well established. However, little is known about the importance of these G1 checkpoint regulators in hyperthermia-induced cytotoxicity. Such information is relevant because of the clinical application of hyperthermia in combination with chemotherapy or with radiotherapy. The effects of p53 or pRb inactivation were studied in a well-established isogenic system using the human colorectal carcinoma cell line (RKO). The cells were treated with clinically relevant heat doses (60 min at 40-43 degrees C). Cell survival, cell-cycle redistribution and induction of apoptosis were investigated. Survival of the p53-inactivated transfectants was higher than that of the wild-type p53 cells. The pRb inactivated transfectants showed an intermediate sensitivity to hyperthermia. All transfectants showed G2 arrest after hyperthermia and the appearance of a sub-G1 population. The induction of apoptosis was inhibited in p53-inactivated and pRb inactivated transfectants. These results suggest that p53 and/or pRb status may be an important determinant of the clinical response to hyperthermia. PMID- 10473868 TI - Basic fibroblast growth factor potentiates cisplatinum-induced cytotoxicity in MCF-7 human breast cancer cells. AB - Basic fibroblast growth factor (bFGF) is a classical mitogen in fibroblasts and endothelial cells. Our previous studies have demonstrated that bFGF inhibits the growth of MCF-7 human breast cancer cells. The aim of the present study was to examine the effect of bFGF on cis-diamminedichloroplatinum(cisplatin)-induced cytotoxicity in MCF-7 breast cancer cells as compared to normal endothelial cells. MCF-7/NCF cells transduced with a vector expressing the bFGF gene and overexpressing its product, and MCF-7/N2 cells transduced with the backbone vector were incubated with a combination of bFGF and cisplatin for 5 days; results were compared with those obtained with bovine aortic endothelial cells. Cell proliferation was assessed with the sulforhodamine B colorimetric cytotoxicity assay. Apoptosis was quantitatively determined by flow-cytometric analysis for DNA damage and the apoptotic death assay for DNA fragmentation, and qualitatively by electron microscopy. Reverse transcriptase/polymerase chain reaction analysis and an enzyme immunoassay were used to determine the mRNA and protein level, respectively, of the anti-apoptotic bcl-2 gene product. We found that bFGF enhanced cisplatin-induced cytotoxicity in MCF-7 breast cancer sublines. bFGF enhanced proliferation of normal endothelial cells and did not increase cisplatin-induced cytotoxicity. This effect was accompanied by down regulation of the anti-apoptotic protooncogene bcl-2 and the enhancement of cisplatin-induced apoptosis. We suggest that the improved understanding of the role of bFGF in the differential modulation of the response of breast cancer and normal endothelial cells to chemotherapy may enable active intervention to alter the therapeutic ratio favorably in breast cancer patients. PMID- 10473869 TI - New potent sensitizers for photodynamic therapy: 21-oxaporphyrin, 21 thiaporphyrin and 21,23-dithiaporphyrin induce extensive tumor necrosis. AB - New sensitizers for photodynamic therapy (PDT) are reported. These compounds, namely 21-thiaporphyrin, 21,23-dithiaporphyrin and 21-oxaporphyrin, reveal some of the properties required for such therapy. Their physicochemical, chemical and pharmacological features meant that we could use them in the treatment of transplantable BFS1 fibrosarcoma in Balb/c mice. New sensitizers and the well known chlorin e6 (Ce6) were used in doses of 2.5, 5.0, 7.5 and 10.0 mg/kg body weight, given intraperitoneally and followed by light irradiation, the total light doses being 50, 100 and 150 J/cm(2) within 24 h after injection. The effectiveness of new sensitizers in PDT was evaluated with in terms of tumor necrosis intensity, the survival time of treated animals, the rate of tumor response (complete/partial/no response), and skin photosensitivity. These results were compared to results obtained in analogous conditions after Ce6-PDT. Distribution studies revealed that the highest concentration of new compounds occurred within 24 h after injection. The results of these experiments confirmed that 21-thiaporphyrin, 21,23-dithiaporphyrin and 21-oxaporphyrin can be considered as potent tumor photosensitizers that do not exert any unwanted effects, primarily skin photosensitization. We suggest that these porphyrins are possible sensitizers to be applied in clinical PDT. PMID- 10473870 TI - Sialyl-Lewis(x/a)-decorated selectin ligands in head and neck tumours. AB - PURPOSE: E- and P-selectins, expressed on vascular endothelium, and their sialyl Lewis(x )(sLe(x))- and/or sialyl-Lewis(a) (sLe(a))-containing ligands have a crucial role in extravasation and metastasis of circulating cells. We wanted to analyse the role of selectins and their ligands in head and neck tumours. METHODS: A total of 40 consecutive biopsy specimens were collected from surgery performed at the Helsinki University Central Hospital between September 1995 and November 1996. The series of specimens contained both benign and malignant head and neck tumours of epithelial, lymphoid or mesenchymal origin. All these were analysed with immunohistochemistry for epithelial and endothelial expression of sLe(x) and sLe(a) glycans and E- and P-selectins. RESULTS: Epithelial expression of sLe(x) and sLe(a) glycans was higher in benign than in malignant lesions in both epithelial and lymphoid tumours. On the other hand, endothelial expression of sLe(x), sLe(a), E- and P-selectin was lower in benign than in malignant lesions in both epithelial and lymphoid tumours. CONCLUSIONS: These data suggest that altered epithelial and endothelial expression of sLe(x) and sLe(a )glycans acting on selectin ligands is linked to the development of head and neck tumours. PMID- 10473871 TI - Serum levels of vascular endothelial growth factor and basic fibroblast growth factor in patients with soft-tissue sarcoma. AB - PURPOSE: Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) have been suggested to be important mediators for tumor-induced angiogenesis. We measured serum VEGF and bFGF levels from patients with soft tissue sarcomas and correlated serum VEGF and bFGF levels with tumor status at surgery and histological grading. MATERIALS AND METHODS: A group of 18 healthy controls and 85 patients with soft-tissue sarcoma were enrolled in this study. The patients were classified according to tumor status at surgery. Serum levels of VEGF and bFGF were also correlated with histological grading. VEGF and bFGF levels were determined by enzyme-linked immunosorbent assay (Quantikine R&D Systems). RESULTS: Serum VEGF and bFGF levels were significantly elevated in the patient group (VEGF: 580pg/ml, bFbF: 21pg/ml, P = 0.0001). The highest concentrations of serum VEGF and bFGF were found in patients with macroscopic tumor lesions or G3 histology. Serum VEGF levels showed a statistically significant correlation with tumor status and grading (P = 0.006 for tumor status, P = 0.0001 for grading). CONCLUSIONS: This study reveals that elevated preoperative serum VEGF and bFGF levels can be detected in the majority of patients with soft-tissue sarcoma. The significant correlation with tumor mass and histological grading suggests that a consecutive monitoring of VEGF and bFGF in the serum of patients with soft-tissue sarcoma might be a valuable marker for tumor follow-up. PMID- 10473872 TI - Anti-T antibodies and peanut-agglutinin-binding glycoproteins in sera of patients with gastric cancer. AB - Agglutinating antibodies to neuraminidase-treated red blood cells (anti-T agglutinins) are known to be reduced in patients with gastric cancer. The antigenic determinant of anti-T agglutinin is known to have a disaccharide structure [Gal(beta1-3)GalNAc], the same specificity as peanut agglutinin (PNA). We examined sera of 27 patients with gastric cancer and 30 controls for anti-T agglutinins, anti-T antibodies and PNA-binding glycoproteins. Anti-T agglutinins were titrated by a microtiter hemagglutination method. Levels of anti-T antibodies were determined by enzyme immunoassay using synthetic glycoconjugate [Gal(beta1-3)GalNAc O-alpha-linked to human serum albumin] as an antigen. Levels of PNA-binding glycoproteins in sera were measured by sandwich enzyme-linked lectin assay using wheat germ agglutinin and peroxidase-conjugated PNA. Titers of anti-T agglutinins were significantly lower in patients with gastric cancer than in controls (P = 0.041). Levels of anti-T antibodies were not significantly different in patients with gastric cancer and controls; however, decreased levels of anti-T antibodies were more frequent in patients with gastric cancer than in controls (P = 0. 001). Levels of PNA-binding glycoproteins were significantly higher in sera of patients with gastric cancer than in controls (P = 0.001). The levels of anti-T antibodies inversely correlated with the levels of PNA-binding glycoproteins in sera of patients with gastric cancer (r = -0.44, P = 0.021). These results suggest that the decrease in anti-T antibodies in sera of patients with gastric cancer might be due to immune complex formation between circulating PNA-binding glycoproteins and anti-T antibodies. PMID- 10473873 TI - IIb/IIIa Receptor Blockers. Preface. PMID- 10473874 TI - Results of IIb/IIIa receptor blockade in patients with unstable angina. AB - The efficacy of GPIIb/IIIa inhibitors has now been evaluated in over 20 000 patients with unstable angina and non-Q MI. These agents have shown great efficacy in patients undergoing percutaneous coronary intervention. They are also effective, even if to a lesser degree, as an addition to medical treatment. The safety profile is satisfactory. Several issues have to be investigated in the future: comparison of agents, use of oral inhibitors, associations with Heparin, consequences on management, and cost efficacy. As of now, it is clear that GPIIb/IIIa inhibitors represent a significant progress in the treatment of patients with acute coronary syndromes. PMID- 10473875 TI - Clinical trials of IIb/IIIa receptor blockers in patients undergoing angioplasty. AB - Many major clinical trials have been published in the last decade involving a new class of antiplatelet agent-Glycoprotein IIb/IIIa receptor blockers. The largest experience to date with these has been in percutaneous coronary intervention. Seven trials involving three different agents (abciximab, tirofiban, eptifibatide) are discussed. Abciximab is the most widely studied agent and is the only drug licensed for use in this setting. All three agents reduce the incidence of clinically relevant ischaemic events (death, non-fatal myocardial infarction or urgent revascularization). The shorter acting, competitive inhibitors tend to be maximally beneficial during the time of infusion, whereas abciximab has been shown to be effective in the acute and long-term phases. The benefits of treatment are tempered by an increase in the bleeding complications. These can be minimized by changes in heparin dosing and careful management of vascular sheaths. The treatment benefit of abciximab is maintained in those patients with unstable angina, those undergoing atherectomy, vein graft angioplasty or bail out-stenting. Results from the Epistent trial, support the use of abciximab during elective stenting. Nevertheless, rapidly changing interventional techniques and the availability of other potent antiplatelet agents underscore the need for further evaluation of IIb/IIIa inhibition in coronary revascularisation. PMID- 10473876 TI - Stents and IIb/IIIa receptor blockers combined: usefulness in various types of coronary artery disease. AB - Both the implantation of intracoronary stents and the use of glycoprotein IIb/IIIa receptor blocking agents have been proven to be of value in the invasive management of coronary artery disease. Stenting is shown to decrease restenosis rate considerably, and is of great use in countering complications of balloon angioplasty, e.g. dissection and abrupt closure. The use of glycoprotein IIb/IIIa receptor blocking agents has been demonstrated to be beneficial as an adjunctive to intracoronary interventions; it has a profound effect on the rate of acute intervention related complications, but not on the occurrence of chronic restenosis. The scope of this article is to evaluate the usefulness of the combination of stents and glycoprotein IIb/IIIa blocking agents in various manifestations of coronary artery disease. It is concluded that a glycoprotein IIb/IIIa receptor antagonist as an adjunctive to the application of a stent is especially useful when thrombus can be assumed to be present in a clinically relevant quantity. PMID- 10473877 TI - The use of platelet glycoprotein IIb/IIIa receptor antagonists during the acute phase of myocardial infarction. PMID- 10473878 TI - The evolution of oral antiplatelet therapy. PMID- 10473879 TI - Safety of the use of IIb/IIIa receptor blockers with or without the use of other anticoagulants. PMID- 10473880 TI - The delta-globin RNA transcript level in beta-thalassemia carriers. AB - Increased levels of hemoglobin A(2) (HbA(2)) are present in most beta-thalassemia carriers. The mechanism of this effect is not understood, although the increase may result from transcriptional and posttranscriptional changes. In the present study, we quantitate delta-globin mRNA levels in peripheral-blood-enriched reticulocytes and characterize the variation of delta-mRNA levels in 30 beta thalassemia heterozygotes who individually carry one of the four common Chinese beta-thalassemia alleles [codons 41/42 (-TTCT); codon 17 (A-->T); IVS-II-654 (C- >T); -28 (A-->G)]. A sensitive and quantitative competitive reverse-transcriptase polymerase chain reaction method was developed and used to assess the absolute amounts of delta-mRNA transcripts in these peripheral erythroid cells. The results showed a large increase in delta-mRNA amounts in all the carriers examined (72.3 +/- 9.0 amol/microg RNA) as compared with those in 12 controls (1.2 +/- 0.2 amol/ microg RNA). There was a direct correlation between the delta mRNA levels and types of beta-thalassemia alleles; generally, the delta-mRNA levels are higher in heterozygotes for beta(0)-thalassemia mutations than beta(+) thalassemia mutations. The delta-mRNA levels correlated inversely with hemoglobin and red cell indices but directly with HbA(2) levels in heterozygotes of each of the group of beta-thalassemia mutations. These results suggest that a greater impairment in beta-globin gene expression results in increased transcription of delta-globin gene and in a higher level of HbA(2). PMID- 10473881 TI - Bacteremia by gram-negative bacilli in patients with hematologic malignancies. Comparison of the clinical presentation and outcome of infections by enterobacteria and non-glucose-fermenting gram-negative bacilli. AB - To compare the characteristics of bacteremic infections by different aerobic gram negative bacilli (GNB) in patients with hematologic malignancies, we studied 54 consecutive monomicrobial bacteremias by Enterobacteriaceae (EB), 15 by Pseudomonas aeruginosa, 43 by other non-glucose-fermenting GNB (NGFGNB) and 11 by other GNB. Patients with EB and P. aeruginosa bacteremia usually developed the infection after intensive chemotherapy for leukemia or during a hematopoietic stem cell transplantation, while most infections in outpatients off therapy were due to NGFGNB. A significant proportion of bacteremias by EB (37%) and P. aeruginosa (40%) were accompanied by severe morbidity (septic shock, pneumonia or deep-seated organ infections) vs. only 7% of other NGFGNB (p < 0.01). Most infections by these latter bacteria were catheter-related bacteremias (80 vs. 2% of EB, p < 0.005), while most EB infections (61%) were uncomplicated bacteremias of unknown source (vs. 14% of other NGFGNB, p < 0.005). Appropriate antibiotics alone cured 98% of EB and 73% of P. aeruginosa bacteremias but only 26% of other NGFGNB (p < 0.005 for both differences), which were cured by catheter removal in 70%, usually after failure of antibiotic treatment. In conclusion, our results suggest that there are significant differences in the patient populations and clinical characteristics of bacteremic infections by the classic GNB (EB and P. aeruginosa) and the new NGFGNB in adults with hematologic malignancies. PMID- 10473882 TI - Response to plasma exchange and steroids as combined therapy for patients with thrombotic thrombocytopenic purpura. AB - We describe our experience in the management of 11 consecutive patients with thrombotic thrombocytopenic purpura (TTP) treated with a combined therapy of plasma exchange (PE) and steroids. Nine patients (82%) achieved complete remission (CR) after a median of 6 rounds of PE (range 2-22). There were 3 early relapses managed in the same way as the initial episode. One patient relapsed 23 months after diagnosis achieving CR with standard therapy; another patient suffered several relapses, and splenectomy was performed after the last one. Three patients died, 2 of them with resistant disease 9 and 38 days after diagnosis, and the remaining one died due to AIDS-related complications while he was in CR. Eight patients are alive in CR with a median follow-up of 38 months (range 8-74). The combination of PE and steroids is a well-tolerated and effective treatment of TTP, but improvements in therapy are needed to manage refractory patients. PMID- 10473883 TI - Sequential use of deferiprone and desferrioxamine in primary school children with thalassaemia major in Turkey. AB - The effectiveness of the sequential use of deferiprone and desferrioxamine (DFO) in children with thalassaemia major was examined. Seven thalassaemic children in whom urinary iron induced by deferiprone was sufficient to maintain a negative iron balance were enrolled in the long-term trial. Deferiprone at a dose of 75 mg/kd/day in 3 divided doses was given for 4 school days a week. The group was given DFO at a dose of 40-50 mg/kg/day s.c. over 8-12 h with a battery-operated pump for 2 days at the weekend. In addition to the safety variables, they were monitored for serum ferritin levels at 2-month intervals and hepatic iron concentrations in liver tissues were determined at the beginning and the 6th month of therapy. The severity of hepatic damage was graded according to the Knodell hepatic activity index and the fibrosis was quantified. None of the patients suffered adverse effects of the therapy but a transient increase in serum ALT levels was noted. A nonsignificant decline in serum ferritin was observed (p = 0.08), a significant reduction in hepatic iron concentration was also determined (p = 0. 03). The hepatic activity index in liver tissues of the patients at the 6th month of the sequential therapy significantly decreased (p = 0.03) whereas fibrosis scores did not differ significantly (p = 0. 25). PMID- 10473884 TI - Establishment and characterization of a Fas-resistant T cell line. AB - Fas is a cell surface receptor that controls a signal transduction pathway leading to apoptosis. We established an antihuman Fas monoclonal antibody (mAb) resistant variant, kit-225-FR, from the human T cell line, kit-225. Flow cytometric analysis revealed that the expression of Fas molecules on kit-225-FR was preserved. The defect in Fas molecule was not detected either by reverse transcription polymerase chain reaction (PCR) analysis of the Fas transcript or by PCR-single strand conformation polymorphism analysis of the Fas gene in kit 225-FR. Although kit-225-FR was resistant to a high concentration of anti-Fas mAb, apoptosis could be induced, as with the wild type, by exogenous C2-ceramide exposure. MORT1/FADD was expressed at wild-type level in kit-225-FR, as determined by Western blot analysis. It therefore appears that the apoptotic signal transduction in kit-225-FR is defective between FADD and the sphingomyelin ceramide pathway. By comparing the differences from the wild-type kit-225, kit 225-FR would serve as a useful cell line for analyzing Fas-specific signal transduction pathways in detail. PMID- 10473885 TI - Levels of endothelial, neutrophil and platelet-specific factors in sickle cell anemia patients during hydroxyurea therapy. AB - It has been shown that the clinical course of sickle cell (SS) patients can be ameliorated by administration of hydroxyurea (HU). Induction of hemoglobin F (HbF) is thought to be the mechanism responsible for clinical improvement in some patients. However, HU has a variable effect on HbF production and there exists no good correlation between the extent of HbF increase and clinical response. On the other hand, the degree of adherence of SS to vascular endothelium and neutrophil counts correlate well with clinical severity. Being a cytotoxic drug, used in myeloproliferative diseases, HU may alter proliferation among various cell lines. Moreover, HU has been reported to reduce red blood cell (RBC) adhesion receptor expression in young SS individuals and induces changes in endothelial cells in vitro. It should be conceived that in addition to its effects on HbF production, HU may change the clinical symptoms of SS patients by affecting the degree of adherence of different blood cells, by influencing the activity of endothelium as well as the activity of white blood cells (WBC) and platelets. To analyze whether several of the determinants of adhesion are modulated by HU treatment we studied the levels of endothelial activity (soluble vascular adhesion molecule-1, (sVCAM 1), interleukin-8 (IL-8), fibronectin, neutrophil activity (sL-selectin, sIL-6 receptor-alpha, myeloperoxidase) and platelet activity (von Willebrand factor) in relation to clinical symptoms, hematological data and HbF levels in 8 SS patients before and during 5 months of HU therapy. Steady state sVCAM-1 levels are increased compared to normal controls and a significant decrease is noted during HU treatment, suggesting a decrease in the interactions between RBC and vascular endothelium. The IL-8 levels are comparable to those in normal controls and remain unaffected by HU therapy. Intercurrent infection and crises reveal striking increases in IL-8 which are accompanied by leukocytosis, but otherwise the IL-8 levels do not correlate with hematological data. HU has no demonstrable effect on fibronectin or soluble neutrophil adhesion molecules, but the levels of myeloperoxidase decrease significantly while WBC counts do not, implying a reduction in neutrophil activity which may help attenuate the propagation phase of a vasoocclusive crisis. PMID- 10473886 TI - Evaluation of the left ventricular systolic and diastolic functions by echocardiography in patients with acute leukemia. AB - Eighteen patients with acute leukemia were studied to evaluate their left ventricular systolic and diastolic function by M mode, two-dimensional and Doppler echocardiographic techniques. The study group was compared with another group of 18 patients of similar age with iron deficiency anemia. The latter group had a comparable heart rate and levels of Hb. There were no differences in systolic function, whereas the diastolic function was lower in patients with acute leukemia (E/A ratio = 1.1 +/- 0.3 vs. 1.42 +/- 0.5, p < 0.05). It is concluded that left ventricular diastolic dysfunction is more common in patients with acute leukemia and may be an indicator of restrictive cardiomyopathy. PMID- 10473887 TI - Factor VIII inhibitors in patients with hemophilia A. AB - In 58 hemophilia A patients aged 1-18 years (mean 9.5 +/- 4.7 years), the prevalence of inhibitors was found to be 27% by the Bethesda method in November 1995. Inhibitor activity was not detected in any of 14 patients with mild hemophilia while it was present in 9 of 27 (33%) patients with moderate, and 7 of 17 (41%) with severe disease. During follow-up, the inhibitors were transient in 10 of 16 patients (17%) and the prevalence of inhibitors was 10% at the end of the study. Our study has demonstrated that the patients' age, factor VIII (F VIII) coagulant activity levels, type of F VIII replacement therapy, and frequency of F VIII administration affect inhibitor development, and these factors should be considered in the follow-up of hemophiliacs. PMID- 10473888 TI - A case of ALK-positive large T-cell lymphoma expressing epithelial membrane antigen with favorable prognosis. AB - We describe the case of a 59-year-old woman with Ki-1-negative, T cell-type, anaplastic lymphoma kinase (ALK)-positive large cell lymphoma that was positive for epithelial membrane antigen. She was histopathologically diagnosed as having a metastatic undifferentiated carcinoma from a cervical lymph node biopsy. Clinical and radiographic studies revealed no other primary tumor. The patient underwent a left radical neck dissection. Surgically resected lymph nodes revealed an ALK1-positive large cell lymphoma. Thereafter the patient has had an unusually favorable prognosis. PMID- 10473889 TI - Shiga toxins do not directly stimulate alpha-granule secretion or enhance aggregation of human platelets. PMID- 10473890 TI - Pilot study of the combination of melphalan, carboplatin and etoposide as a conditioning regimen for relapsed lymphoma patients. PMID- 10473891 TI - Proliferating cells in biopsy sections of leukemic bone marrow. PMID- 10473892 TI - Brainstem auditory evoked response recorded using maximum length sequences in term neonates. AB - Brainstem auditory evoked response (BAER) was studied in term neonates using maximum length sequences (MLS), which allows presentation of acoustic stimuli at much higher rate than possible with conventional averaging technique. The response waveforms, especially for waves III and V, were well defined at the click rates of 91-455/s. Wave latencies and interpeak intervals at various click rates were all significantly longer and wave V amplitude was smaller in the neonates than in the adults. As click rate was increased, wave latencies increased and amplitudes progressively reduced. The slopes of wave latency-rate functions were progressively steeper for the later waves. The slopes of both III V and I-V interval-rate functions were significantly steeper in the neonates than in the adults, indicating greater rate-dependent changes in the central components of the neonatal MLS BAER. These rate-dependent changes in the neonatal MLS BAER are generally similar to those in neonatal conventional BAER, although there are some differences. The results indicate that the neonatal brainstem can process rapid acoustic stimulation up to 455/s, although this ability remains immature. PMID- 10473893 TI - Cerebrospinal fluid and serum nitric oxide levels in asphyxiated newborns. AB - Hypoxic-ischemic encephalopathy (HIE) is the result of a chain of events caused mainly by cytokines and nitric oxide (NO) release, which is later on followed by free oxygen radical injury. To investigate NO involvement in asphyxiated newborns, serum and cerebrospinal fluid (CSF) values of NO levels in 17 neonates with HIE were detected. Infants at or above 37 weeks of gestation were classified to have mild, moderate and severe HIE due to Sarnat and Sarnat. Samples obtained between 24 and 72 h of life were immediately frozen at -70 degrees C till the time of measurement by Sievers NOA. Five patients had mild, 6 patients had moderate and 6 patients had severe HIE, 4 in the severe HIE group also had multisystem involvement. The CSF NO levels were significantly higher in moderate and severe HIE groups compared to the mild HIE group (p = 0.028 and p = 0.018 respectively). Our results show that NO level increases in CSF with the severity of HIE between 24 and 72 h following asphyxia. According to the animal work, this is the time period where inducible NO synthase gets activated and could cause neurotoxicity, which might perhaps be prevented by interventions. PMID- 10473894 TI - Effect of doxapram on episodes of apnoea, bradycardia and hypoxaemia in preterm infants. AB - AIM: To study the effect of doxapram on the frequency of apnoea, bradycardia and hypoxaemia. METHODS: Fifteen infants, median gestational age at birth 27 weeks (range 24-30), age at study 27 days (12-60), with >/=6 episodes of bradycardia or hypoxaemia/6 h despite serum caffeine levels in the therapeutic range, received doxapram either intravenously (0.5-2 mg/kg/h) or orally (2-8 mg/kg every 2 h). Six-hour recordings of pulse oximeter saturation (S(P)O(2)), pulse waveforms, ECG, breathing movements and nasal airflow were performed immediately before as well as 1, 3 and 6 days after onset of treatment. Recordings were analysed for apnoea (>/=4 s), bradycardia (heart rate < 2/3 of baseline) and hypoxaemia (S(P)O(2) 1 kHz) where thresholds differed by up to 50 dB. These differences showed no apparent correspondence to the ability to produce sounds (vocal versus non-vocal species) or to the spectral content of species-specific sounds. All fishes have maximum sensitivity between 400 Hz and 1,500 Hz, whereas the major portion of the energy of acoustic signals was in the frequency range of 100-400 Hz (swimbladder drumming sounds) and of 1-3 kHz (stridulatory sounds). Species producing stridulatory sounds exhibited better high-frequency hearing sensitivity (pimelodids, doradids), except for callichthyids, which had poorest hearing ability in this range. Furthermore, fishes emitting both low- and high-frequency sounds, such as pimelodid and doradid catfishes, did not possess two corresponding auditory sensitivity maxima. Based on these results it is concluded that selective pressures involved in the evolution of the Weberian apparatus and the design of vocal signals in otophysines were others (primarily predator or prey detection in quiet freshwater habitats) than those serving to optimize acoustical communication. PMID- 10473906 TI - CSF monoamines, age and impulsivity in wild grivet monkeys (Cercopithecus aethiops aethiops). AB - Brain monoaminergic activity has been associated with behaviors, such as impulsive risk-taking, that tend to peak during adolescence in humans and nonhuman primates. This study was designed to assess natural variation in monoamine neurotransmitter metabolism in relation to age and behavioral impulsivity in grivet monkeys (Cercopithecus aethiops aethiops) living in their native habitat and subject to natural ecological pressures. Cisternal cerebrospinal fluid, collected from 22 animals living in the Awash National Park, Ethiopia, was assayed for the major metabolites of serotonin (5 hydroxyindoleacetic acid, 5-HIAA), dopamine (homovanillic acid, HVA) and norepinephrine (3-methoxy-4-hydroxyphenylglycol, MHPG). Concentrations of HVA declined significantly from one year of age to older adulthood. Further, a significant curvilinear relationship was identified between age and the 5 HIAA/HVA ratio, with the trough coinciding with the period of adolescence. Finally, behavioral impulsivity, as measured by re-entering baited traps a second time after the animal had already been captured and sampled for CSF, was related to lower levels of MHPG. The results suggest that normal variation in central monoaminergic activity may have functional consequences in wild populations. PMID- 10473907 TI - Clopidogrel for cerebrovascular prevention. AB - Ischemic stroke, myocardial infarction and peripheral arterial disease are different clinical manifestations commonly due to the same underlying disease, i.e. atherosclerosis with subsequent thrombosis/embolism (atherothrombosis). Many clinical trials of secondary prevention after stroke or TIA have evaluated the benefit of long-term use of antiplatelet drugs in reducing the risk of subsequent vascular events. Aspirin and triclopidine have been shown to be effective in placebo-controlled studies for the composite outcome of stroke, myocardial infarction, or vascular death. Contrasting with these benefits, there were potentially serious, though rare, adverse effects. These considerations certainly justify the development of new antiplatelet agents. Clopidogrel is a new ADP receptor antagonist, with a greater activity in animal models of thrombosis. CAPRIE (Clopidogrel versus Aspirin in Patients at Risk of Ischemic Events) was a randomized, blinded, international trial designed to assess the relative efficacy of clopidogrel and aspirin in reducing the risk of the outcome cluster of ischemic stroke, myocardial infarction, or vascular death, as well as to assess their relative safety. 19,185 patients were recruited. The intention-to-treat analysis showed that the relative risk reduction was 8.7% (95% CI 0.3-16.5, p = 0.043) in favor of clopidogrel from an overall annual event rate of ischemic stroke, myocardial infarction, or vascular death, ranging from 5.83% in the aspirin group to 5.33% in the clopidogrel group. The percentage of adverse events reported was higher in the aspirin group for all categories except rash, diarrhea, and abnormal liver function. It seems likely that clopidogrel will replace ticlopidine for stroke prevention, because of its better safety profile, and comparable efficacy. Clopidogrel probably will not replace aspirin as the first line therapy for many clinicians because of its higher cost and lack of widespread experience. However, other clinicians have already decided that they will use clopidogrel as first choice, because of the significant advantage over aspirin demonstrated in the CAPRIE study. PMID- 10473908 TI - Temporary occlusion of middle cerebral artery by macroembolism in carotid surgery. AB - Two patients are presented who during carotid endarterectomy (CEA) temporarily showed an obstruction of the middle cerebral artery (MCA) mainstem by a macroembolus resulting in cerebral ischaemia. Both cases are unusual examples of CEA and selected from a cohort of more than 1,500 operations. During surgery with general anaesthesia, brain function was monitored with computerized electroencephalography (EEG) and transcranial Doppler (TCD) ultrasonography. The simultaneous use of EEG and TCD monitoring allowed us to witness the development of intraoperative cerebral ischaemia and to relate these events to a temporary occlusion of the MCA mainstem by a macroembolus. This is the first life report that describes obstruction of a cerebral artery by arterial embolism resulting in cerebral ischaemia. PMID- 10473909 TI - Cerebral microembolus generation in different extracorporeal circulation systems. AB - Microemboli generated during extracorporeal circulation (ECC) are likely to induce neurological sequelae. This study examines whether the choice of a distinct type of ECC can reduce intracerebral emboli counts. Middle cerebral artery blood flow during coronary artery bypass grafting was monitored continuously by transcranial Doppler ultrasound in 45 patients. The ECC systems used were a roller pump (n = 16), a centrifugal pump (n = 18) and a combination of centrifugal pump and heparin-coated ECC system (n = 11). Patients' characteristics as well as surgical and anesthesiological procedure did not differ between the groups. Total counts did not differ significantly between the three groups. Intraoperative events in individual patients may lead to massive embolus generation overcoming positive properties of a distinct ECC system. PMID- 10473910 TI - Emergency carotid endarterectomy. AB - OBJECTIVE: Evaluation of the therapeutical efficacy of emergency carotid endarterectomy (CEA) in neurologically unstable patients. PATIENTS AND METHODS: Three groups of a consecutive series of 71 emergency CEAs performed from 1980 to July 1998 were classified: (1) acute onset of severe stroke (n = 16), (2) progressive stroke/stroke in evolution (n = 34), and (3) crescendo transient ischemic attacks (n = 21). Cerebral coma, cerebral haemorrhage, and major ischemic stroke established in cranial computed tomography scans were contraindications for surgery. The neurological outcome was assessed by the modified Rankin scale. Long-term survival and long-term stroke recurrences were analyzed. RESULTS: The recovery/minor stroke rates (Rankin 0-3) in acute stroke, progressive stroke, and crescendo transient ischemic attacks were 56.3, 76.4 and 80.9%, respectively; the combined major stroke/mortality rates (Rankin 4-6) were 43.7, 23.6 and 19.1%, respectively. Intraoperative angiography in 39 patients detected early carotid reocclusions in 2 and intracranial embolism in 7 patients. Local application of thrombolytic agents (n = 5) may contribute to a better neurological outcome in emergency CEA. Life table probabilities of major strokefree survival were 74.5, 71.6, and 53.7% after 1, 2, and 5 years, respectively (including perioperative strokes). Life table probabilities to suffer no stroke recurrence during follow-up were 96.7, 96.7 and 85.3%, respectively (perioperative strokes excluded). CONCLUSIONS: Emergency CEA may be worthwhile in selected patients. Completion angiography is mandatory. Emergency CEA should be included in therapeutic strategies for ischemic stroke. PMID- 10473911 TI - Clinical features and prognosis in young adults with infratentorial infarcts. AB - BACKGROUND AND PURPOSE: Many comprehensive descriptions of the clinical spectrum of infratentorial infarcts in elderly patients and with a retrospective design have been published. The aim of this study was to describe the clinical characteristics and prognosis in young patients with isolated infratentorial infarcts. METHODS: In a prospective series of 105 patients aged 18-44 years with cerebral infarction 24 had a brainstem or cerebellar infarction. The patient selection was validated in a population-based epidemiological survey. The patients were assessed acutely and at 4 and 12 months after onset. Extensive evaluation included CT and MRI scans, angiography, ultrasonic duplex scanning, transesophageal echocardiography and a chemistry panel including hematologic testing. The modified Rankin scale and NIH stroke scale were used for assessment of disability and neurological dysfunction. RESULTS: Eighteen patients had a cerebellar infarct (posterior inferior cerebellar artery territory in 9 patients, superior cerebellar artery in 6, anterior inferior cerebellar artery in 2, nonterritorial in 1). Two patients had lateral medullary infarcts and 2 isolated pontine infarcts. In 2 patients MRI was normal despite repeated investigations. Hearing loss and tinnitus were the only explicit symptoms for superior cerebellar artery infarcts, but it was otherwise impossible to classify each case to a vascular territory according to clinical characteristics. The age-specific incidence of isolated cerebellar infarction was 1.8/100,000/year. The presumed causes were arterial dissection in 8 patients, idiopathic in 7, cardioembolic in 5, oral contraceptive use in 3 and protein S deficiency in 1 patient. One patient died during the acute phase and another developed a locked-in syndrome. At follow up, 1 patient had a transitory ischemic attack and 1 a silent cerebral infarction. Twenty-two patients had a favorable outcome according to the modified Rankin scale (grade 0-2) and the NIH scale. CONCLUSIONS: Cerebellar infarctions are frequent among young stroke patients in northern Sweden. Arterial dissection is the prevailing stroke mechanism in infratentorial infarcts. The prognosis is favorable regarding motor impairment but cognitive deficits may prevent return to work. PMID- 10473912 TI - Noninvasive assessment of the circle of Willis in cerebral ischemia: the potential of CT angiography and contrast-enhanced transcranial color-coded duplexsonography. AB - Thirty-four patients with acute hemispheric ischemic strokes underwent both CT angiography and contrast-enhanced transcranial color-coded duplexsonography (TCCD) to study the effectiveness of the combined noninvasive techniques for evaluation of the circle of Willis. In 3/34 patients, CT angiography and contrast enhanced TCCD demonstrated middle cerebral artery (MCA) occlusion, in 5 others MCA stenosis. A severe posterior cerebral artery stenosis was missed by CT angiography. In 8 patients, contrast-enhanced TCCD failed because of poor bone windows. In these patients, CT angiography was normal. CT angiography and contrast-enhanced TCCD are complementary noninvasive diagnostic tools. Disagreements between the diagnostic findings of these methods still need further evaluation by digital subtraction angiography. PMID- 10473913 TI - Cerebral vein thrombosis in a case with thromboangiitis obliterans. AB - Thromboangiitis obliterans is a chronic inflammatory vessel disease that involves predominantly the small and medium-sized arteries and veins of the distal extremities. Appearance and cessation of symptoms are closely related to patterns of tobacco consumption. That cerebral arteries can also be involved is shown by reports of rare cases in which cerebral artery occlusion led to infarction. We report on a 28-year-old man with thromboangiitis obliterans who developed extensive cerebral vein thrombosis after a single episode of cigarette smoking following several years of nonsmoking. Despite extensive evaluation, no other known cause or predisposition of cerebral vein thrombosis could be found. This case suggests that cerebral veins can be involved in thromboangiitis obliterans and patients with thromboangiitis obliterans might be at risk for cerebral vein thrombosis. PMID- 10473914 TI - Cerebral thrombophlebitis in three patients with probable multiple sclerosis. Role of lumbar puncture or intravenous corticosteroid treatment. AB - We report 3 cases of young patients, 2 women and 1 man, who presented a cerebral venous thrombosis following intravenous treatment with high doses of corticosteroids. All of them presented a probable multiple sclerosis according to clinical, biological (CSF) and MRI criteria and were treated for the first time by a bolus of 1,000 mg of methylprednisolone OD during 5 days. All the usual causes of cerebral venous thrombosis were systematically excluded in all of them. The role of corticosteroid treatment in cerebral thrombophlebitis is discussed. All of them underwent a lumbar puncture a few days before corticosteroid treatment and the relationship between lumbar puncture and cerebral thrombophlebitis is also discussed. Cerebral venous thrombosis associated with corticosteroid treatment has rarely been reported. The relationship between corticosteroids and venous thrombosis has already been suggested but has never been clearly understood. PMID- 10473915 TI - Measures to improve recovery in the acute phase of stroke. PMID- 10473916 TI - Recovery from aphasia and neglect. PMID- 10473917 TI - Functional imaging of recovery from stroke: A review of personal experience. PMID- 10473918 TI - Pharmacological approaches to stroke recovery. PMID- 10473919 TI - Systemic fungal infections in patients with hematologic malignancies: indications and limitations of the antifungal armamentarium. AB - The rates of fungal infections have increased substantially in Europe as well as in North America. Most frequently Aspergillus spp. and Candida spp. are isolated. Despite the recent introduction of new azoles and lipid-based formulations of amphotericin B, there are relatively few randomized, controlled studies on the use of antifungal drugs in patients with hematological malignancies and invasive fungal infections. Conventional amphotericin B is considered the gold standard for the treatment of invasive fungal infections; however, adverse events limit conventional amphotericin B treatment. The newer azoles, fluconazole and itraconazole, are well tolerated; however, fluconazole has no activity against Aspergillus spp. An additional serious problem is the emerging resistance of nonalbicans Candida spp. to fluconazole. In this situation, lipid formulations of amphotericin B seem to be attractive, since the use of these drugs has been shown to be safe and effective. Considerably higher medical costs limit broader application of lipid formulations of amphotericin B. Because of the rapidly increasing incidence of serious fungal infections, we have reviewed current strategies and the role of newer antifungal drugs for the treatment of deep-organ infections. PMID- 10473920 TI - Susceptibility surveillance among gram-negative bacilli at a cancer center. AB - We conducted a survey of susceptibility among 758 gram-negative bacilli (GNB; collected from cancer patients over a 3-month period) to commonly used antibiotics. The overall resistance among GNB was least for piperacillin/tazobactam and meropenem (5 and 6%, respectively) followed by cefepime (8%), imipenem (9%), amikacin (12%), ofloxacin (13%), ciprofloxacin, ceftazidime and ticarcillin/clavulanate (14% each), aztreonam (18%) and tobramycin (24%). In comparison to data on antibiotic resistance to ceftazidime, imipenem, ciprofloxacin and aztreonam in similar studies in 1985 and 1994, resistance has significantly increased to all four antibiotic classes. Based on our current study, meropenem, cefepime, imipenem and piperacillin/tazobactam would be the most appropriate choices in our institution for empiric therapy of GNB infections in febrile neutropenic patients. PMID- 10473922 TI - In vitro bactericidal activities of antimicrobial agents and morphologic changes on Prevotella bivia. AB - Prevotella bivia is common in pelvic inflammatory diseases. Parenteral antimicrobial agents have been widely used against those infections. We investigated the bactericidal activities of three cephalosporins, i.e. cefluprenam (CFLP), ceftazidime (CAZ) and cefotaxime (CTX) and of two other antimicrobial agents, i.e. clindamycin (CLDM) and imipenem (IPM) against P. bivia. We also investigated the in vitro morphological changes induced by these agents in P. bivia. Cephalosporins exhibited bactericidal activities against P. bivia and induced time- and concentration-dependent morphological changes in P. bivia (filamentation). CLDM and IPM also had bactericidal activities, but induced different morphologic alterations: formation of spheroblasts and lysis. These results confirm the fact that each antimicrobial agent has characteristic aspects. PMID- 10473921 TI - 'Subinhibitory' erythromycin represses production of Pseudomonas aeruginosa lectins, autoinducer and virulence factors. AB - Pseudomonas aeruginosa infection is preceded by selective adhesion of the bacteria to the host target cells via diverse adhesins, including lectins. This step enables maximal damage to the target host cells by the bacterially secreted injurious toxins and enzymes. The production of both lectins and many of the virulence factors is positively controlled by transcription activators including signaling autoinducers (N-acyl-L-homoserine lactones). We show in this communication that erythromycin at subminimal growth inhibitory concentrations simultaneously suppresses the production of P. aeruginosa hemagglutinins (including lectins), protease, hemolysin and homoserine lactone autoinducers. The antibiotic-treated bacteria also show reduced virulence to mice, endorsing clinical observations that indicate the efficiency of low-dose erythromycin treatment of persistent drug-resistant P. aeruginosa infections. PMID- 10473923 TI - A cluster of nosocomial cross-infection due to multiple antibiotic-resistant Acinetobacter baumannii: Characterization of the strain and antibiotic susceptibility studies. AB - A multiple antibiotic-resistant (MAR) strain of Acinetobacter baumannii caused nosocomial cross-infection among 3 patients of a surgical intensive care unit. The isolates were of identical biochemical profile (77776 S-U-) and serotype (serovar 36) and identical in terms of pulsed-field gel electrophoresis macrorestriction (SmaI, ApaI) analysis. This MAR strain was susceptible only to netilmicin, tobramycin, imipenem, meropenem, polymyxin B, and trovafloxacin. The minimal bactericidal concentrations of imipenem and meropenem were markedly higher than the corresponding minimal inhibitory concentrations against this strain. Combined fresh defibrinated human blood (65 vol%) and antimicrobial drug assays yielded the following results: polymyxin was the most rapidly bactericidally effective antibiotic in the presence of blood and in broth. Tobramycin and netilmicin were efficacious in 65 vol% blood. Imipenem was slightly more effective than meropenem in broth, whereas both carbapenems sterilized blood-containing assay tube contents. Trovafloxacin failed to achieve bactericidal activity (to 99.9% kill) in the presence of blood, presumably because this strain was resistant to ciprofloxacin and borderline susceptible to ofloxacin. Trovafloxacin combined with either imipenem or meropenem yielded an indifferent effect. However, the combination of trovafloxacin (2 microg/ml) plus tobramycin (1 microg/ml) achieved sterilization of tube contents in the presence of blood within 4 h after exposure and in broth following extended (overnight) incubation. This MAR strain of A. baumannii was high-level resistant to rifampin; thus the combination of polymyxin B plus rifampin proved indifferent. PMID- 10473924 TI - On the inhibitory activities of a new boron compound and ultrasound against the mouse ascites tumour. AB - The inhibitory effects of a new boron compound, dihydroxy (oxybiguanido) boron (III) hydrochloride monohydrate (HB), and ultrasound (US) of a frequency 25 kHz on the growth of ascites tumour in female Swiss mice were studied by monitoring the survival, weight of tumour-associated material, tumour cell count, serum alkaline phosphatase activity and the haematological parameters of the treated animals. 5-Fluorouracil (5-FU), a well-known anticancer agent, was used as positive control. While HB exhibited a very significant antitumour action, US alone produced a small but significant inhibitory effect. The combination of US with HB or 5-FU produced an extra antitumour action as compared to the actions of these chemicals used singly. The mechanisms of action of the new boron compound (HB) and US are discussed. PMID- 10473925 TI - A comparative study of cefepime versus ceftazidime as empiric therapy of febrile episodes in neutropenic patients. AB - An open-label, randomized comparative study was conducted to evaluate the efficacy and safety of cefepime (2.0 g q. 8 h) and ceftazidime (2.0 g q. 8 h) in the empiric therapy of febrile neutropenic patients. A total of 45 eligible febrile episodes were randomized (1:1) to be treated with the study regimen. Nineteen febrile episodes treated with cefepime and 22 febrile episodes treated with ceftazidime were evaluable for efficacy. The two groups were comparable in terms of age, sex, height, weight, underlying neoplasm, number of pretherapy neutrophil, duration of neutropenia and types of infections. The overall therapeutic success rate of the cefepime group (53%) was comparable to the ceftazidime group (50%). It did not differ significantly (95% confidence interval: -0.28 to 0. 34, p = 0.85). Eighty-eight percent of pathogens in each group were bacteriologically eradicated. The safety profile was similar in both groups. No patients in either group discontinued the therapy because of adverse events. None (0%) of the cefepime patients and 2 (9%) of the ceftazidime patients died of infection. The results of this study suggest that cefepime is an effective and safe agent in the empiric therapy of febrile episodes in neutropenic patients. PMID- 10473926 TI - A prospective, randomised comparison of single- vs. multiple-dose antibiotic prophylaxis in penetrating trauma. AB - This study was designed to compare the clinical efficacy of a single dose of ceftriaxone with cefoxitin given 3 times a day for 3 days. METHODS: Patients had to have a penetrating injury to only one part of the body, reach the hospital within 2 h and be operated on within 16 h after the trauma. Patients were excluded if it appeared likely that they would require mechanical ventilation for more than 24 h. The same applies to open or grade II/III craniocerebral trauma. The end point was the occurrence of infections within 10 days. The costs of antibiotic treatment were also calculated. RESULTS: 96% of the ceftriaxone patients (n = 97) and 95% of the cefoxitin group (n = 98) remained infection free. In neither treatment group was deep infection, abscess, phlegmon or sepsis seen. No additional surgery or intensive care due to infection was required. At $41.83 vs. $172.16, the average total cost of delivering antibiotic treatment was significantly lower in the ceftriaxone group (p < 0.001). CONCLUSION: Prophylaxis in penetrating trauma with a single dose of ceftriaxone is safe and has considerable practical and economic advantages. PMID- 10473927 TI - An adverse interaction between warfarin and 5-fluorouracil: A case report and review of the literature. AB - Adverse interactions between warfarin and 5-fluorouracil (5-FU) have been reported. Such an interaction occurred in a patient with lung cancer receiving vinblastine and 5-FU. This case is the first involving a patient taking minidose warfarin for prophylaxis of catheter-associated thrombosis. Although the mechanism of the interaction is unclear, it has been postulated that 5-FU interferes with the synthesis of hepatic cytochrome P-450 2C9. Because warfarin and 5-FU are regularly coadministered, this adverse interaction might be occurring more frequently than is realized. Clinicians should be aware of this interaction and should regularly monitor the prothrombin time of patients receiving warfarin and 5-FU. PMID- 10473928 TI - Adequate macrolide treatment schedules for uterine cervicitis caused by Chlamydia trachomatis. AB - To evaluate in vivo efficacy of macrolides; erythromycin (EM) and clarithromycin (CAM), in the different treatment schedules of Chlamydia trachomatis uterine cervicitis im women. Cervical C. trachomatis was detected by the polymerase chain reaction. EM was orally administered to 8, 15 and 18 Japanese patients, at a dosage of 600 mg, t.i.d. for 5, 7 and 14 days, respectively. CAM was orally administered to 10, 26 and 19 Japanese patients at a dosage of 400 mg, b.i.d. for 5, 7 and 14 days, respectively. The eradication rate and the recurrence rate in the different treatment schedules of C. trachomatis were evaluated. The eradication rates after 5, 7 or 14 days of treatment with EM were 1/8, 8/15 or 13/18, respectively. The recurrence rates after 5, 7 or 14 days of treatment with EM were 1/1, 3/8 or 2/13, respectively. On the other hand, the eradication rates after 5, 7 or 14 days of treatment with CAM were 5/10, 26/26 or 19/19, respectively. The recurrence rates after 5, 7 or 14 days of treatment with CAM were 3/5, 0/26 or 0/19, respectively. A 7-day treatment with CAM would be adequate and effective for C. trachomatis uterine cervicitis. PMID- 10473929 TI - The concept of delirium. AB - The concept of delirium has a long and confusing history. This article outlines the development of ideas relating to core features of the syndrome: disturbance of consciousness, disturbance of cognition, its course and its external causation. The modern concept of delirium, and the diagnostic criteria found in current classifications are based upon a long tradition of clinical observation in younger patients, and their emphasis on positive symptoms and identifiable external causes may not be applicable to our ageing population. PMID- 10473930 TI - Epidemiology of delirium. AB - Delirium is one of the most frequent symptoms of disease in the elderly. A large variation of incidence and prevalence data is reported probably due to different patient populations and inconsistent diagnostic criteria. In medical and surgical elderly inpatients recent studies report a prevalence rate of approximately 15% and in postoperative patients the incidence and prevalence rates vary greatly, 7 52%, depending on patient population and clinical setting. In nursing homes the prevalence is even higher and delirium is often combined with dementia. Data supports the statement that delirium is most often found in hospitalized somatically ill elderly patients. As a consequence of the rising number of elderly in hospitals we have to expect an increase in the prevalence and incidence of delirium. PMID- 10473931 TI - Do delirium and Alzheimer's dementia share specific pathogenetic mechanisms? AB - Dementia is the most common risk factor for delirium in the elderly. Here we will review the evidence that proposed pathogenetic mechanisms for delirium (such as reduced cerebral metabolism, imbalance of the noradrenergic/cholinergic neurotransmission, inflammation, disturbances in neuronal systems which regulate stress and the sleep/wake cycle) are also a part of the pathophysiology of Alzheimer's disease. In particular, the role of inflammatory mechanisms in both disorders will be discussed. PMID- 10473932 TI - Clinical profile of delirium in patients treated for femoral neck fractures. AB - The incidence of delirium, its predisposing factors, clinical profile, associated symptoms and consequences were investigated in 54 consecutive patients, 19 men and 35 women, mean age 77.1 years, admitted to an 'ortho-geriatric unit' with femoral neck fractures. The incidence of postoperative delirium was 15/54 (27.8%) and a logistic regression model found that dementia and a prolonged waiting time for the operation increased the risk of postoperative delirium. Delirium during the night was most common but in 5 patients the delirium was worst in the morning. Patients with delirium suffered more anxiety, depressed mood, emotionalism, delusions and hallucinations. A larger proportion of patients with delirium could not return to their previous dwelling, and a larger proportion of delirious patients were either dead, wheelchair-bound or bedridden at the 6-month follow-up (p < 0.005). The conclusion is that delirium is common and has a serious impact on the outcome after hip fracture surgery. PMID- 10473933 TI - Update on the neuropathogenesis of delirium. AB - Delirium has been considered a syndrome of generalized dysfunction of higher cortical functions due to its breadth of symptoms and associated diffuse slowing on electroencephalogram. Advances in neuropsychiatry have revealed differences between brain regions, including the hemispheres, which may underlie the constellation of symptoms among different psychiatric disorders. For example, different neural pathways are involved in major depression and obsessive compulsive disorder, including lateralization to one or the other hemisphere. In this article the author proposes that delirium, too, involves particular neural pathways and that lateralization to the right may be relevant. Structural and functional neuroimaging reports and recent neuropsychological studies support this lateralization. Prefrontal cortices, anterior and right thalamus, and right basilar mesial temporoparietal cortex may play a significant role in subserving delirium symptoms and may be the 'final common pathway' for delirium from a variety of etiologies. The final common pathway may be responsible for certain 'core symptoms' (disorientation, cognitive deficits, sleep-wake cycle disturbance, disorganized thinking, and language abnormalities), while other symptoms (delusions, hallucinations, illusions, and affective lability) may occur depending on the etiology causing delirium. An imbalance in the cholinergic and dopaminergic neurotransmitter systems is most commonly implicated in causing delirium, and could both account for delirium symptoms and be consistent with the neuroanatomical pathways being implicated. PMID- 10473934 TI - Cerebrometabolic aspects of delirium in relationship to dementia. AB - Delirium is associated with decreased cerebral metabolism and with cholinergic deficiency. These deficits also occur in Alzheimer's disease, the most common form of dementia. The clinical, metabolic, and pharmacological similarities between delirium and dementia agree with the suggestion that delirium and dementia can both be thought of as forms of 'cerebral insufficiency'. PMID- 10473935 TI - Neurochemistry and delirium. AB - Acute confusional state or delirium is one of the most common organic brain syndromes especially in the elderly. It develops suddenly, within hours or days, and often during a period of hospitalization. In this paper we review the results of our studies on the neurochemistry of delirium. PMID- 10473936 TI - Acetylcholine and delirium. AB - The neurotransmitter acetylcholine has been implicated in animal and human studies of delirium. This chapter will briefly review the clinical studies focussing on measurement of serum levels of anticholinergic activity in delirious states. Three approaches have been taken. First, to identify medications currently prescribed that have subtle anticholinergic effects. The current 'list' includes 48 commonly prescribed medications. Second, to associate serum anticholinergic activity with delirium in various clinical states including postcardiotomy delirium, postelectroconvulsive delirium, delirious elderly medical inpatients, and nursing home patients. Third, to intervene in patients with elevated anticholinergic activity by reducing known anticholinergics and correlating this reduction with clinical measures of cognition and delirium. Our most recent data investigates the impact of anticholinergics on demented patients. Rates of delirium were significantly higher in patients receiving larger numbers of anticholinergics. PMID- 10473937 TI - Activity in the hypothalamic-pituitary-adrenal axis and delirium. AB - Glucocorticoid hormones are important for coping with stress but may have deleterious effects on mood and memory during prolonged excessive secretion. A key abnormality related to cortisol excess in delirium seems to be abnormal 'shut off' of the hypothalamic-pituitary-adrenal (HPA) axis tested by the dexamethasone suppression test. In experimental models, the hippocampal formation is of prime importance for normal HPA axis shut-off. In this brain area, a close interaction between neurotransmittors, notably acetylcholine, serotonin and noradrenaline, and glucocorticoid receptors, is present and possibly relevant for the development of delirium in elderly patients with stroke and neurodegenerative brain diseases. PMID- 10473938 TI - Social and environmental contributants to delirium in the elderly. AB - Even if the mechanisms and etiology of delirium still require further clarification, this article works on the assumption that delirium is caused by stress factors acting on the brain to such an extent that the brain can no longer cope. Therefore, social and environmental factors involving high levels of stress will cause delirium in the elderly. The influence of these factors is obviously very complex with several potential interacting mechanisms between different factors. Since studies on this subject are limited to certain aspects of the complex topic discussed, only narrowly focussed systematic knowledge on this topic is available. However, it can be concluded that hospital admission for example is an important risk factor for developing delirium. PMID- 10473939 TI - Identifying and recognizing delirium. AB - Clinicians frequently under-recognize delirium. Physicians and nurses can be taught to recognize delirium by focusing on each of the components of the criteria for delirium. Teachers of physicians and nurses should emphasize the ability to recognize acuteness of onset, fluctuation of course, and attention deficits. Attention deficits in the clinical setting can be recognized by digit span testing, spelling words backwards, or assessing accessibility during the clinical interview. Attention can be scored by assigning a score from 0-10, or ranking as mild, moderate or severe, the patients ability to engage in the medical interview. PMID- 10473940 TI - The diagnosis of delirium and DSM-IV. AB - This paper reviews the history of the development of the American Psychiatric Association's Diagnostic and Statistical Manual (DSM) using the diagnostic category of delirium as an example. It also attempts to show how clinical practice and experience has molded the current conceptualization of delirium in DSM-IV. The paper presents the process and options the DSM-IV Work Group on Organic Disorders pursued in developing the DSM-IV criteria for the diagnosis of delirium, particularly in relation to ICD-10. PMID- 10473941 TI - What criteria should be used for the diagnosis of delirium? AB - DSM-IV made several revisions to the diagnostic criteria for delirium. This paper reviews the empirical data that supported these revisions. More research is needed to evaluate the usefulness of the new criteria and to further define subtypes of delirium. PMID- 10473942 TI - Assessment scales in delirium. AB - In research and in clinical practice instruments assessing symptoms of delirium are of value in the diagnostic process, in the evaluation of intervention as well as in the care of patients. Different assessment instruments are needed for different purposes. Most instruments developed are screening instruments or diagnostic tools. Confusional State Evaluation (CSE) is an observer's rating scale assessing severity of delirium. Interrater reliability and validity of the CSE have been found satisfactory. The CSE seems to be of value in following the course of delirium in elderly patients and useful for instance in evaluating effects of interventions. PMID- 10473943 TI - Clinical subtypes of delirium in the elderly. AB - Psychomotor disturbance is common in delirium, with some patients being restless and hyperactive and others lethargic and hypoalert. Although patients with hyperactive delirium may be recognised more readily, hypoactive and mixed forms of delirium are more common on general hospital wards. Recent evidence suggests that hyperactive delirium has a better prognosis than other subtypes. It remains uncertain whether this reflects fundamental differences in the pathophysiology of different subtypes. PMID- 10473944 TI - Can delirium be separated from dementia? AB - This presentation reviews the nosological status of delirium and dementia as a prelude to a discussion of how to distinguish them, or even if they are distinguishable. It argues that committee (ICD, DSM) classification systems fossilise the meaning of the syndromes, and thereafter have a baleful effect on research. The distinction between dementia and delirium is a case in point, and it can be argued that this is much less clear than we thought. It is suggested that validation of syndromes as prognostic tools is a logical way out of the nosological swamp into which delirium and dementia research is sinking, and that cognitive improvement should be the key distinction. PMID- 10473945 TI - Delirium in clinical practice: experiences from a specialized delirium ward. AB - Geriatric patients with known dementia and suffering from an acute somatic disease are highly vulnerable to develop delirium. It is therefore essential to suspect and recognize delirium in these patients, especially in emergency wards. In the present study we evaluated activities on a dedicated delirium ward at a Swedish University Hospital. Over one and a half years 637 patients were treated for suspected delirium, the majority of patients being referred from the emergency ward at the same hospital. Infectious diseases were the main cause of delirium in 67% of cases. Other common causes were heart disease and stroke. Drug use as the only cause of delirium was found in less than 1% of cases. Approximately 70% of patients had cognitive disturbances, either dementia or mild cognitive impairment. The existence of multiple diseases as causative factors was frequent. Knowledge about delirium and how it is both diagnosed and treated is of great importance in all kinds of settings where acute somatic treatments are common. PMID- 10473946 TI - Predisposing and precipitating factors for delirium in hospitalized older patients. AB - Delirium is a common and serious problem for older hospitalized patients. This investigation proposes a multifactorial model of delirium etiology, involving a complex interrelationship of predisposing (vulnerability) factors and precipitating factors (acute insults). An overview of risk factors for delirium identified in 14 studies published since 1980 is provided. Although these studies identify key risk factors for delirium, they do not allow the examination of the interrelationship of predisposing and precipitating factors. Thus, we present two prospective cohort studies by our group which empirically examine: (1) predisposing (vulnerability) factors, (2) precipitating factors, and (3) the interrelationship of predisposing and precipitating factors. Understanding these risk factors is the key to developing appropriate preventive strategies and to target intermediate and high risk patients for intervention efforts. PMID- 10473947 TI - Postoperative delirium. AB - This article reviews the incidence, pathophysiological hypotheses, and etiology of postoperative delirium, especially in the elderly. Preoperative, intraoperative, and postoperative risk factors for delirium following surgery are discussed. Results of various studies on postoperative delirium appear hardly comparable due to methodological and population differences. Therefore, it is difficult to draw any conclusions on postoperative delirium in general. Special attention is paid to delirium after cardiac surgery since this syndrome has been studied most. PMID- 10473948 TI - Delirium: effectiveness of systematic interventions. AB - The purpose of this paper was to review evidence related to the effectiveness of systematic interventions in preventing or detecting and treating delirium. The review process involved a systematic search of the literature on each topic, assessment of the validity of the studies retrieved and examination of their results. The literature search identified 10 studies on prevention and 7 studies on detection and treatment. A broad spectrum of systematic interventions appeared to be modestly effective in preventing delirium in young and old surgical patients but not elderly medical patients. Systematic detection and treatment programs and special nursing care appeared to add large benefits to traditional medical care in young and old surgical patients and modest benefits in elderly medical patients; however, it seemed that the more precise the target of the detection and treatment program, the greater the benefit. PMID- 10473949 TI - Drugs that induce delirium. AB - Drug-induced delirium is often seen in clinical practice. Most knowledge about delirium-inducing drugs comes from reports on side effects, and few systematic studies have been done in the field. Delirium is strongly associated with anticholinergic activity; drugs of different classes, including tricyclic antidepressants and traditional high-dose neuroleptics, constitute a high-risk group. A large number of drugs, including benzodiazepines, sedatives, dopamine activating drugs, antiepileptics, histamine H2 receptor blockers, digitalis and analgesics, are less frequently associated with delirious reactions and constitute a medium-high-risk group. Some of these drugs do not have anticholinergic effects but in vitro have shown to bind to muscarine receptors. The risk of inducing delirium in frail elderly and demented persons clearly suggests that drugs which might induced delirium should be avoided. PMID- 10473950 TI - Care of the delirious patient. AB - In spite of the fact that delirium is a common and often severe cognitive disturbance in the elderly, quite few intervention studies are performed. Descriptive studies of variable quality are much more common. For example, in hip fracture patients delirium is a common complication and the cause of nursing problems that cannot be explained by the fracture per se. Nursing and medical interventions have been published separately but only one study, the Pitea Program, has so far been known that combines nursing and medical knowledge. This program has been shown to reduce the incidence of delirium in elderly hip fracture patients. PMID- 10473951 TI - Impact of delirium on professionals. AB - There is a paucity of empirical research on the impact of delirium on professionals' reactions, interpretations or attitudes to people in a delirious state. This is striking because it stands to reason that it is demanding to take care of a person who is disoriented, perhaps agitated and who may hallucinate and/or act in a manner that calls for interventions involving the use of force or actions that oppose the will of the patient. Apart from some recent intervention studies, there is also little empirical research focusing on the care of delirious patients. Because of the lack of research, deductions will be made from research findings in other areas as well as research findings focusing on the views of patients about what it was like to have been confused, which they narrated afterwards. PMID- 10473952 TI - Educational interventions in delirium. AB - In the absence of specific therapies, interventions for delirium are largely educational. This paper reviews educational interventions targeted at physicians, nurses, patients and their families. Most studies to date have had methodological deficiencies, and few have been developed in keeping with current principles for adult learning. Early studies focused on increased recognition of delirium, with less consistent measurement of patient centred outcomes. Subsequent studies provide limited evidence to suggest that, as a result of educational interventions, individual symptoms can be prevented, and, when delirium is present, its consequences - including death and prolonged hospital stays - can be diminished. Future studies should incorporate principles of adult learning and have improved methodological rigour. The strength of educational interventions needs to be assessed carefully. Interventions which are time-sensitive, less complex, directed to observable, patient-focused endpoints are more likely to be successful. PMID- 10473953 TI - What are natural retinoids? AB - Vitamin A (retinol) is the prototype of all other natural retinoids. It is composed of a nonaromatic ring structure, a polyprenoid side chain and a carbonyl end group. These features make it liable to metabolic interconversion and protein interactions but also cause detergent-like properties and a sensitivity to UV irradiation and oxidation. Natural retinoids are present in all living organisms, either as preformed vitamin A or as carotenoids, and are required for a vast number of biological processes, e.g. vision, cellular growth and differentiation and reproduction. Although retinol is the most omnipotent compound, natural retinoids like all-trans-, 9-cis- and didehydroretinoic acid (RA) are clearly more potent outside the retina and trigger gene expression via binding to nuclear retinoid receptors. Retinaldehyde takes an intermediate position in this respect, with ability to convert to both retinol and RA. Over the years, many natural retinoids have been tried therapeutically against skin disorders with the best effects achieved with retinol, retinaldehyde, 13-cis-RA and all-trans-RA. The latter compound was the prototype when new, synthetic derivatives of vitamin A were sought, hoping for a better therapeutic index and a higher functional specificity. Inevitably, treatment with such drugs will influence the effects of coexisting natural retinoids. An understanding of the basic principles of these interactions may have major impact on patient outcome. PMID- 10473954 TI - Metabolism of topical retinaldehyde. AB - OBJECTIVE: In order to circumvent the tolerance problems encountered with topical application of retinoic acid - a biologically active metabolite of vitamin A - we performed in various models a series of experiments aimed at assessing the bio availability of topical retinaldehyde and its conversion into either retinoid stores or biologically active metabolites. METHODS: (i) (3)H-retinaldehyde was used as a precursor of either (3)H-retinol or (3)H-retinoic acid in human skin extracts and human cultured keratinocytes; (ii) the concentration of various retinoids resulting from the metabolism of topical retinaldehyde was determined in mouse skin and in human plasma. Retinoids were quantified by reverse-phase HPLC with UV detection. RESULTS: Human keratinocytes were shown to take up retinaldehyde and to convert it into retinoic acid in a differentiation-dependent manner, differentiating cells oxidising retinaldehyde more efficiently. In vivo models allowed us to demonstrate that retinaldehyde is taken up by the skin and is then predominantly converted into retinyl esters - a storage form of vitamin A - while delivering relatively low amounts of retinoic acid from a large reservoir. CONCLUSION: Topical retinaldehyde can be used as a precursor of endogenous retinoids, since it is converted into both storage and bio-active forms of vitamin A. PMID- 10473955 TI - Biological activities of topical retinaldehyde. AB - BACKGROUND: We had hypothesised that retinaldehyde (RAL) should be an interesting precursor for topical use. AIM: We review our observations about its biological activities. METHODS: We performed pilot studies to explore its biological effects and tolerability in human skin and compared the effects of topical RAL to that of all-trans-retinoic acid (RA) in the mouse tail test. RESULTS: The biological activities of RAL were found to be qualitatively identical to that of RA: (i) induction of cellular RA-binding protein type 2 mRNA and protein, (ii) increase in epidermal proliferation (increase in DNA synthesis, epidermal thickness, induction of 50-kD keratin mRNA and reduction in 70-kD keratin mRNA), and (iii) metaplastic effects (induction of orthokeratosis, reduction of 65-kD keratin mRNA, increase in filaggrin and loricrin mRNAs). When associated with RAL, citral (known for its capacity to inhibit the oxidation of retinol to RA in epidermis) counteracted the effects induced by RAL indicating that RAL exerts biological activities through transformation to RA. Hypothesizing that keratinocytes would metabolize 9-cis-RAL to 9-cis-RA, we compared the biological effects induced by topical 9-cis-RAL and found that hyperplastic and metaplastic responses were lower than those induced by all-trans-RAL or all-trans-RA at similar concentrations. This suggests that 9-cis-RAL has no advantage over all-trans-RAL for specific delivery of natural retinoids into the skin. As in clinical studies conducted in human skin, we also found topical RAL less irritant than RA. CONCLUSION: These studies indicate that topical RAL has biological activity and is well tolerated. PMID- 10473956 TI - Inhibitory effects of retinoids on vascular endothelial growth factor production by cultured human skin keratinocytes. AB - BACKGROUND: Vascular endothelial growth factor (VEGF), a potent angiogenic factor and vasodilator, is strongly expressed by epidermal keratinocytes in many angiogenesis-dependent skin disorders. Retinoids may modulate VEGF in skin and this may be related to an effect on rosacea. AIM: To investigate the effect of retinaldehyde on VEGF production by human keratinocytes. METHODS: The effects of different concentrations of retinoids (all-trans-retinal and all-trans-retinoic acid) on VEGF production by cultured human skin keratinocytes in both cell extracts and supernatants were determined. Expression of VEGF was analyzed by enzyme-linked immunosorbent assay (ELISA) and RT-PCR. RESULTS: The amount of cell associated and secreted VEGF strongly decreased with retinoid concentration (e.g. 48, 69% inhibition at 0.1 microM all-trans-retinal and -retinoic acid, respectively, in the supernatants). In parallel, approximately 25% inhibition of VEGF mRNA expression was obtained in the presence of 0.01 microM all-trans retinal. CONCLUSION: The decrease in VEGF expression by keratinocytes on contact with retinoids may prevent skin neoangiogenesis in certain skin diseases. PMID- 10473957 TI - Antibacterial activity of retinaldehyde against Propionibacterium acnes. AB - BACKGROUND: Retinaldehyde has been shown to exert antibacterial activity in vitro. AIM: This study evaluates the effect of retinaldehyde on Propionibacterium acnes both in vivo and in vitro. METHODS: Microbial minimal inhibitory concentrations (MICs) of retinaldehyde and retinoic acid were determined on reference strains of P. acnes. In vivo activity of daily topical application of 0.05% retinaldehyde on the P. acnes density was evaluated after application in a single-blind randomised study. RESULTS: MICs of retinaldehyde were 4 mg/l for P. acnes No. CIP179 and CIP53119 and 8 mg/l for P. acnes No. CIP53117. In contrast, the MICs of retinoic acid were superior to 128 mg/l for these three strains. In vivo, retinaldehyde-treated areas displayed a significant decrease in counts of viable P. acnes as compared with the untreated areas with a median decrease of 10(2) log P. acnes/cm(2) after 2 weeks of daily application. Vehicle alone had no effect. CONCLUSION: The MIC of retinaldehyde against P. acnes suggests a direct antibacterial activity. Daily topical application of 0.05% retinaldehyde is associated with a clear reduction of the P. acnes density. PMID- 10473958 TI - Comedolytic effect of topical retinaldehyde in the rhino mouse model. AB - BACKGROUND: Retinaldehyde is a key molecule in the metabolism of vitamin A by keratinocytes. In order to evaluate its range of topical activity in acne, its comedolytic effect was compared to that of retinoic acid in the same vehicle, in the rhino mouse model. METHODS: The animals were treated on the back daily for 5 consecutive days per week for 3 weeks. At the end of this period, histological slides were analyzed in order to quantify the features of comedones and epidermal thickness. RESULTS: Topical treatment with a retinaldehyde (0.05% w/w) and a retinoic acid formulation (0. 025% w/w) induced comedolysis and increased the epidermal thickness with the same intensity. CONCLUSION: These data indicate that retinaldehyde exerts a significant comedolytic activity. PMID- 10473959 TI - Efficacy of topical 0.05% retinaldehyde in skin aging by ultrasound and rheological techniques. AB - BACKGROUND: The natural precursor of retinoic acid, i.e. retinaldehyde, has been proven to exert retinoid activities. AIM AND METHODS: The aim of this prospective instrument study was to determine the effect of topical retinaldehyde 0.05% on the physical properties of aging skin. This was performed using two devices, namely a high-resolution (70-80 microm) ultrasound scanner, which visualizes the thickness of both the epidermis and the dermis, and an echorheometer, which assesses the stiffness and elasticity of the skin by suction. In a 1-year study, 21 patients applied retinaldehyde cream 0.05% on the face, while another group of 19 volunteers were only treated with an emollient (control group). Epidermal and dermal thicknesses were measured on the forehead and temple, and stiffness and elasticity were measured on the forehead only. All the instrumental parameters were assessed at baseline and at the end of treatment. RESULTS: Compared to the control group, retinaldehyde treatment induced a significant increase in epidermal thickness of the temple, as well as in cutaneous elasticity (p < 0. 01). Similarly, retinaldehyde treatment tended to increase dermal thickness and reduce cutaneous stiffness, but no statistical difference could be observed between the two groups. CONCLUSION: Taken together, the results further suggest that retinaldehyde has counteracting effects on skin aging PMID- 10473960 TI - Repair of UVA-induced elastic fiber and collagen damage by 0.05% retinaldehyde cream in an ex vivo human skin model. AB - BACKGROUND: Cellular effects of UV exposure are implicated in cutaneous aging. UV radiations induce structural and cellular changes in all the compartments of skin. AIM: To study the antiaging efficacy of a cream containing 0.05% retinaldehyde with an ex vivo technique using human skin in order to approximate in vivo metabolic conditions. METHODS: Human skin explants were maintained alive in organ culture for 18 days and subjected to UVA exposure, thus simulating skin photoaging. Retinaldehyde cream was then applied to the surface of the epidermis for 2 weeks and the results were compared with those of nontreated skin explants. Dermal repair was analyzed histologically with quantification of collagen and elastic fibers, and biochemically by the measure of newly synthesized collagen as shown by adding tritiated proline to the culture medium. RESULTS: UVA exposure induced significant alterations of collagen and elastic fibers as shown by morphometric analysis. In all UVA-exposed and then retinaldehyde-treated skin specimens, collagen and elastic fibers were restored to the level of nonexposed skin. UVA exposure induced a decrease in collagen synthesis, whereas in retinaldehyde-treated UVA-exposed skin the synthesis was similar to that of unexposed skin. CONCLUSION: It has been shown that retinaldehyde has many of the properties of tretinoin in its biological and beneficial effects on photoaging. We have verified some of these previous observations, especially on dermal connective tissue, by obtaining significant repair of elastic fibers and collagen alteration induced by UVA exposure. PMID- 10473961 TI - Clinical use of topical retinaldehyde on photoaged skin. AB - BACKGROUND: Retinaldehyde, the natural precursor of retinoic acid, should exert similar effects on photoaged skin. OBJECTIVE: To establish the efficacy and safety of topical retinaldehyde on photoaged skin. METHODS: Open and controlled clinical studies using image analysis of silicone skin replicas. RESULTS: Retinaldehyde proved efficient and safe. CONCLUSION: Retinaldehyde is efficient and well tolerated for the improvement of the signs of photoaging. PMID- 10473962 TI - Retinaldehyde alleviates rosacea. AB - BACKGROUND: Anecdotal observations suggest that retinoic acid may be effective in mild rosacea. AIM: Our aim was to investigate, by an exploratory clinical and instrumental study, the effects of a topical formulation with the retinoic acid precursor retinaldehyde, in patients with vascular signs of facial rosacea. METHODS: Female patients were treated with a 0.05% retinaldehyde cream that was applied once daily for 6 months. Clinical assessments of persistent erythema and telangiectasia were performed every month, using a 4-point severity score (absent to severe). The clinical response for each parameter was defined as a decrease of at least 1 grade in the severity score. In addition, erythema was further evaluated by measurement of the a* parameter, using a spectrophotometer on lesional and nonlesional areas. RESULTS: A total of 23 women comprised the study population. At baseline, 10 patients had diffuse erythema, 3 patients had isolated telangiectasia and 10 patients had both. During retinaldehyde treatment, a clinical response was revealed in about 75% of the patients with erythema, after 5 months (p < 0.05). Similarly, isolated telangiectasia responded to retinaldehyde, although to a lesser extent and after a longer period of treatment (46% responders after 6 months, nonsignificant). Using the spectrophotometer, the a* parameter diminished in patients with erythema by about 15%, after 2 months of treatment (p = 0.001). CONCLUSION: This study indicates that retinaldehyde has beneficial effects on the vascular component of rosacea. PMID- 10473964 TI - Tolerance of topical retinaldehyde in humans. AB - BACKGROUND: Retinaldehyde (RAL) has been used as a topical agent in many countries since 1994. AIM: To review current data on the tolerance of retinaldehyde and to report the results of a long-term pilot study. METHODS: Data from published and on-file studies have been compiled. Forty-five patients who had applied RAL on the face for 12-89 months were specifically examined for side effects. RESULTS: Studies in humans demonstrated an excellent tolerance of topical RAL on human skin. It was much better tolerated than retinoic acid and could be used even on sensitive facial skin. It does not have phototoxic or photo allergic properties. No side-effects were associated with long-term use. CONCLUSION: Current data indicate a good topical tolerance of RAL in humans. PMID- 10473963 TI - Tolerance profile of retinol, retinaldehyde and retinoic acid under maximized and long-term clinical conditions. AB - BACKGROUND: Topical retinoic acid (RA) causes irritation of the skin. To prevent this side effect, natural precursors of RA have been proposed. The aim of the present study was to compare the local tolerance profiles of retinol (ROL), retinaldehyde (RAL) and RA. METHODS: ROL, RAL and RA were studied using repeated insult patch tests for 14 days (n = 6). Similarly, RAL and RA were assessed in long-term clinical use for 44 weeks (n = 355). Clinical scoring on irritation, measurement of transepidermal water loss (barrier function) and laser Doppler blood flow perfusion units (irritation) were performed. RESULTS: Under maximized conditions, an equally low irritation potential for ROL and RAL and a more pronounced irritant effect with RA could be demonstrated clinically (p < 0.05 in the intergroup analysis). Furthermore, RAL and RA induced more scaling than ROL (p < 0.05), and ROL and RA tended to induce more burning/pruritus than RAL (nonsignificant). The TEWL values were low with ROL and high with RAL and RA (nonsignificant, intergroup analysis). The laser Doppler measurements confirmed pro-irritating effects of RA and the nonirritating effects of ROL and RAL (p = 0. 001, intergroup analysis). The long-term clinical study showed that the study population developed a high frequency of erythema (44% of the population), scaling (35%) and burning/pruritus (29%) with RA in the first 4 weeks of treatment, whereas these 3 parameters were significantly less frequent with RAL (p < 0.0001 in the intergroup analysis). CONCLUSION: The natural retinoids ROL and RAL do have a good tolerance profile, in contrast with the irritating potential of RA. PMID- 10473965 TI - Does cimetidine improve prospects for cancer patients?. A reappraisal of the evidence to date. AB - BACKGROUND: Evidence first appeared in 1988 that cimetidine as an adjuvant may improve the survival of severely ill gastro-intestinal cancer patients when given peri- or postoperatively. Since then, several studies have appeared which suggest an anticancer activity for cimetidine, although few attempts have been made to corroborate their findings in large, placebo-controlled, double-blind studies. METHOD: We reviewed the literature concerning cimetidine's potential anticancer activity, particularly with regard to gastro-intestinal cancers. RESULTS: Most studies suggest that cimetidine may improve the outcome in cancer patients by a three-pronged mechanism involving (1) inhibition of cancer cell proliferation; (2) stimulation of the lymphocyte activity by inhibition of T cell suppressor function, and (3) inhibition of histamine's activity as a growth factor in tumours. CONCLUSION: Bearing in mind the experimental evidence, as well as the potential and excellent safety profile of cimetidine, more studies are required and justified to clarify cimetidine's protherapeutic activity. PMID- 10473966 TI - Prokinetics in patients with gastroparesis: a systematic analysis. AB - BACKGROUND: The relative potency of prokinetics in patients with gastroparesis has not been systematically studied. This study, therefore, aimed to assess the available data and to compare the effects of different prokinetics on symptoms and gastric emptying rates in patients with gastroparesis. METHODS: A systematic search of the literature was performed, covering the period 1980 to March 1998. All identified studies were reviewed and the following data elements assessed: characteristics of study populations; sample sizes; treatment regimes; drug doses; study design; main outcome variables, and the validity of measurements. RESULTS: In 36 studies, 514 patients were treated with prokinetics p.o. Most studies had methodological limitations (i.e. nonvalidated measurement of symptoms or unblinded treatment). The mean improvement in gastric emptying and the reduction in the symptom score was higher in the open trial group than in patients treated double-blind. Overall, erythromycin seems to have had the strongest effect on gastric emptying as compared to domperidone, cisapride or metoclopramide. Concerning gastrointestinal symptoms, the symptom scores appeared to improve more during treatment with erythromycin than with domperidone, metoclopramide or cisapride. CONCLUSIONS: Most of the available trials have methodological limitations; this limits the conclusions. However, the data suggest that the motilin-agonist erythromycin is superior with regard to the acceleration of gastric emptying, while both erythromycin and domperidone appear to be the most effective with regard to improvements in the symptom score. Additionally, there is a lack of association between changes in gastric emptying times and improvements in symptoms. PMID- 10473967 TI - Expression of vesicular monoamine transporters in endocrine hyperplasia and endocrine tumors of the oxyntic stomach. AB - BACKGROUND: Gastric enterochromaffin-like (ECL) cells selectively express the vesicular monoamine transporter (VMAT) VMAT2, and enterochromaffin (EC) cells the VMAT1 isoform. AIMS: We investigated whether VMAT isoform selection indicates the origin of endocrine hyperplasia and neoplasia from oxyntic ECL or EC cells and may be of prognostic significance in different types of gastric carcinoids. METHODS: Tissue from patients with chronic atrophic gastritis (CAG), Zollinger Ellison-syndrome (ZES), gastric carcinoids and neuroendocrine carcinoma (NEC) was investigated by immunohistology and in situ hybridization. RESULTS: Endocrine cells forming diffuse, linear, and micronodular hyperplasia in CAG and ZES, as well as oxyntic microcarcinoids expressed both VMAT2 and chromogranin A (CgA) but neither VMAT1 nor serotonin. In five of six sporadic carcinoids VMAT2 and CgA but not VMAT1 were detected. One carcinoid was copositive for VMAT1 and serotonin but negative for VMAT2. Electron microscopy confirmed the VMAT2-positive tumors as ECLoma and the VMAT1-immunoreactive carcinoid as EComa. CONCLUSIONS: VMAT2 and VMAT1 are reliable markers for differentiation of gastric endocrine hyperplasia and neoplasia from ECL and EC cells, respectively. The significance of VMAT2 and VMAT1 as prognostic markers lies in the relatively poor prognosis for EComa compared to ECLoma, characterized by VMAT2 positivity. The absence of both VMAT2 and VMAT1 in NEC may indicate poor prognosis. PMID- 10473968 TI - Beta-microseminoprotein in gastric carcinoids: a marker of tumour progression. AB - Gastric carcinoid disease may have a highly varying clinical course depending on the malignancy of the tumour. Many biochemical markers, such as peptides and biogenic amines, have been found in carcinoid tumour tissue but none has been reported to be useful as a predictor of the degree of malignancy of the carcinoid. beta-Microseminoprotein is a small disulphide-rich protein with unknown function present in the secretions on most mucosal surfaces in the body, including the stomach where it is also found in some endocrine cells. We have studied beta-microseminoprotein by immunohistochemistry in the tumour tissue of 29 patients with gastric carcinoid disease. Beta-Microseminoprotein was present in the tumour tissue in 62% of the patients and its presence was correlated to tumour diameter and tissue invasion depth. The presence of beta microseminoprotein in tumour tissue was corroborated by in situ hybridisation. All 4 patients with the solitary sporadic type of tumour and all 6 patients with metastasis had positive immunostaining of the tumour tissue. The serum concentration of beta-microseminoprotein, measured by radioimmunoassay, was increased in all but 2 of 13 patients with gastric carcinoid disease. To a large part the increase was due to the concomitant atrophic corpus gastritis. We conclude that beta-microseminoprotein in tumour tissue is a marker of tumour progression and that measurement of beta-microseminoprotein in serum is less informative than immunohistochemistry. PMID- 10473969 TI - The antacid Talcid adsorbs and neutralizes all proteins secreted by H. pylori including VacA cytotoxin: a new mechanism for its ulcer-healing action? AB - BACKGROUND AND AIM: Helicobacter pylori culture supernatant containing VacA cytotoxin significantly inhibits gastric cell proliferation and delays healing of experimental gastric ulcers. Since cell proliferation is crucial for ulcer healing, the removal of inhibitory effects of H. pylori secreted cytotoxin would have a beneficial effect on the healing process. In this study, we determined whether the antacid Talcid can adsorb, remove, or neutralize H. pylori derived VacA cytotoxin responsible for the above deleterious actions. METHODS: Supernatants of viable H. pylori isogenic strains producing VacA cytotoxin [VacA(+)] and with disrupted cytotoxin gene not producing cytotoxin [VacA(-)] were incubated with either placebo, Talcid 10 mg/ml, omeprazole 10 mg/ml (positive control) for 1-24 h. Treated supernatants were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis to evaluate proteins. We also studied the effect of supernatants on epidermal growth factor stimulated Kato III cell proliferation using BrdU labeling. RESULTS: Talcid very effectively removed from the H. pylori culture supernatant the approximately 90 kD VacA(+) cytotoxin at 3 and 24 h (99.5% removal vs. placebo-treated control; p<0.001). It also removed all other proteins, including 66-kD urease and 58-kD heat shock protein, secreted by both VacA(+) and VacA(-) H. pylori strains. Omeprazole was completely ineffective in this regard. Preincubation with Talcid completely abolished the inhibitory effect of VacA(+) H. pylori culture supernatant on epidermal growth factor stimulated Kato III cell proliferation. CONCLUSION: Adsorption and neutralization by Talcid of all H. pylori secreted proteins may explain, at least in part, the ulcer-healing action of this drug. PMID- 10473970 TI - A new simple immunoassay for detecting Helicobacter pylori infection: antigen in stool specimens. AB - BACKGROUND/AIM: Several diagnostic tests are available for evaluating Helicobacter pylori (Hp) infection: histological examination, culture of gastric biopsy specimens, rapid urease test, urea breath test and serology. A recently marketed direct enzyme immunoassay (HpSA) detects Hp antigen in stool samples. The aim of our study was to evaluate overall diagnostic sensitivity, specificity and positive and negative predictive values of this new diagnostic test. METHODS: We included in the study 84 patients (39 males and 45 females; mean age 49.57 years) with dyspeptic symptoms who were examined by upper gastrointestinal endoscopy. Exclusion criteria were previous treatment with proton pump inhibitors, bismuth compounds or antibiotics. During the endoscopic examination biopsies were taken from antrum and corpus for Hp culture and histological examination, and stool specimens were submitted to the laboratory to be stored until the HpSA test. Hp was judged to be present when culture or histology and culture were positive. The (13)C-urea breath test was done only in culture negative patients in whom either histology or immunoassay or both were positive. RESULTS: Hp was found in 55 patients by both culture and histology. Stool antigen has been detected in 54 of the 55 Hp-positive patients, giving a sensitivity of 98.2% and a negative predictive value of 96.4%. In 2 out of 29 patients HpSA gave a positive result, but the biopsy-based methods were negative, resulting in a low rate of false-positives, with 93.1% specificity and 96.4% positive predictive value; the (13)C-urea breath test confirmed these results as negative. CONCLUSION: Our results show that this new test is highly sensitive and specific for the detection of Hp infection, and it is satisfactorily reproducible. PMID- 10473971 TI - Cigarette smoke increases apoptosis in the gastric mucosa: role of epidermal growth factor. AB - BACKGROUND/AIMS: Apoptosis is a common mechanism for the regulation of cell loss. It is associated with both tissue atrophy and metaplasia. Cigarette smoking has tremendous adverse effects on the stomach and also increases the risk of gastric cancer. This action may be through the change in apoptosis in the stomach. The aim of this study was to examine the effect of cigarette smoking on apoptosis in the gastric mucosa and the possible role of epidermal growth factor (EGF) in this action. METHODS: Gastric blood flow was assessed by the laser Doppler technique. Serum and gastric mucosal EGF levels were measured by RIA. Gastric mucosal apoptosis was determined using TdT-mediated dUTP-biotin nick end labeling (TUNEL). RESULTS: Cigarette smoke exposure decreased serum EGF which was accompanied by a reduction in gastric blood flow. Meanwhile, gastric mucosal cell apoptosis was increased. Administration of EGF (20 microg/kg i.v.) before each cigarette smoke exposure reversed these actions. Removal of salivary glands induced similar effects on the gastric blood flow, apoptosis, and serum EGF level as with cigarette smoke exposure. CONCLUSION: A reduction in serum EGF was involved in the decrease in gastric blood flow and increase in gastric mucosal apoptosis caused by cigarette smoking. PMID- 10473972 TI - Ultra-high-dose lanreotide treatment in patients with metastatic neuroendocrine gastroenteropancreatic tumors. AB - BACKGROUND: Symptomatic control and occasionally even tumor regression of functional neuroendocrine tumors (NET) of the gastroenteropancreatic (GEP) system can be achieved by somatostatin analogues. Assuming a dose-dependent antiproliferative effect of somatostatin analogues, we performed a study with the somatostatin analogue lanreotide in ultra-high dosages in patients with progressive, metastatic GEP NET. PATIENTS AND METHODS: 30 patients with metastatic GEP NET, progressive during treatment with somatostatin analogues (< or =1.5 mg/day) and/or interferon-alpha, underwent ultra-high-dose lanreotide therapy (5 mg lanreotide s.c. three times a day). Tumor growth was evaluated every 3 months. Serum chromogranin A, serum serotonin as well as urinary 5 hydroxyindoleacetic acetic acid levels were also determined at 3-month intervals. In patients with functional tumors, tumor-related symptoms were documented. RESULTS: After a 1-year treatment period with ultra-high-dose lanreotide, 1 complete and 1 partial remission were observed in patients with functional midgut NET. Eleven patients had stable disease and 11 patients showed continuing tumor growth after 3-12 months of treatment. Symptoms decreased significantly during therapy. CONCLUSIONS: Our data show that ultra-high-dose lanreotide treatment in patients with metastatic GEP NET can lead to control of both symptoms and proliferation in at least some patients refractory to conventional therapies. PMID- 10473973 TI - Genomic imprinting of IGF-II and H19 in adult human pancreatic tissues. AB - BACKGROUND/AIM: Genomic imprinting is a chromosomal modification causing differential expression of maternal and paternal alleles. Loss of imprinting (LOI) of IGF-II and H19 has been suggested to be an early oncogenic event in cancerogenesis. Aim of the present study was to describe the status of IGF-II and H19 imprinting in adult human pancreatic tissues. METHODS: Allele-specific gene expression was studied using RNA and DNA from human pancreatic cancer, chronic pancreatitis, and normal pancreas tissues heterozygous for ApaI (IGF-II) or RsaI (H19) restriction fragment length polymorphism. Reverse-transcriptase polymerase chain reaction products were digested with either ApaI or RsaI and analyzed on agarose gels to study the status of allelic expression. The expression level of H19 and IGF-II was studied on Northern blots or by polymerase chain reaction. RESULTS: H19 was imprinted in normal pancreas and in chronic pancreatitis. H19 LOI was observed in 1 of 4 informative cancer tissues and was not associated with increased H19 transcript levels. Biallelic expression of IGF-II was found in 6 of 10 informative cancer tissues and in 6 of 9 informative normal tissues. In chronic pancreatitis, the IGF-II gene was imprinted in all informative samples. IGF-II mRNA was not overexpressed in the tissues showing LOI. CONCLUSION: Low frequencies of H19 LOI and the lack of correlation between biallelic expression and overexpression observed for both H19 and IGF-II suggest that LOI of H19 and IGF-II is not a relevant oncogenic factor during human exocrine pancreatic cancerogenesis. PMID- 10473975 TI - Primary neuroendocrine tumors of the cystic duct. AB - Primary neuroendocrine tumors of the cystic duct are extremely rare. Only 4 cases have been described to date. We report 2 patients in whom a primary neuroendocrine tumor of the cystic duct was incidentally detected during histological examination following cholecystectomy. With regard to the primary neuroendocrine tumor both patients were asymptomatic at the time of diagnosis. However, histologic examination did not confirm that the neuroendocrine tumors had been completely removed. Both patients underwent a second procedure. They are well after 47 and 49 months, respectively. PMID- 10473974 TI - Role of endothelins in trinitrobenzene sulfonic acid-induced colitis in rats. AB - To determine the role of endothelins (ET) on experimental colitis, following intracolonic trinitrobenzene sulfonic acid administration, rats were given orally either bosentan (BS), a nonselective ET receptor antagonist (100 mg/kg in 5% arabic gum), or arabic gum by gavage for 2 or 14 days. Macroscopic damage scores obtained in the vehicle (1.4+/-0.4), acute (4.8+/-0.6) and chronic (3.8+/-0.3) colitis groups were significantly higher than in the control group (0). BS treatment reduced the scores in both acute (3+/- 0.5) and chronic (2.3+/-0.5) colitis groups. Myeloperoxidase (MPO) activities of colonic tissues were elevated in acute and chronic colitis groups (325.1+/-44.9 and 431.8+/-54.6 U/g wet weight) as compared with the control group (73.6+/-11 U/g wet weight). Plasma protein oxidation levels were found to be significantly increased in the chronic colitis group (1,158.1+/-63.4 nmol/ml) compared with the control, ethanol and acute colitis groups (274.3+/-23.1, 490+/-52.2 and 422.2+/-50.5 nmol/ml). BS treatment significantly reduced both the protein oxidation level (375.5+/-46.9 nmol/ml) and MPO activity (167.5+/-35.8 U/g wet weight). The results of the present study suggest the involvement of ETs in the pathogenesis of colonic injury in this animal model of colitis. PMID- 10473976 TI - Ampullary hamartoma: A rare cause of biliary obstruction. AB - Tumors of the papilla of Vater are very rare. Despite advanced imaging techniques the distinction between benign and malignant tumors remains very difficult. Because most ampullary and periampullary tumors are malignant, primary management is surgical. Here we report the case of a 65-year-old man with biliary obstruction caused by an ampullary hamartoma simulating cancer. The correct diagnosis was not established until surgery. PMID- 10473977 TI - White matter changes in stroke patients. Relationship with stroke subtype and outcome. AB - White matter changes (WMC), detected by imaging techniques, are frequent in stroke patients. The aim of the study was to determine how WMC relate to stroke subtypes and to stroke outcome. We made a systematic Medline search for articles appearing with two of the following key words: either 'WMC or white matter lesions or leukoencephalopathy or leukoaraiosis' and 'stroke or cerebral infarct or cerebral hemorrhage or cerebrovascular disease or transient ischemic attack (TIA)'. WMC, as defined radiologically, are present in up to 44% of patients with stroke or TIA and in 50% of patients with vascular dementia. WMC are more frequent in patients with lacunar infarcts, deep intracerebral hemorrhages, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy and cerebral amyloid angiopathy. After an acute ischemic stroke, WMC are associated with a higher risk of death or dependency, recurrent stroke of any type, cerebral bleeding under anticoagulation, myocardial infarction, and poststroke dementia. WMC in stroke patients are often associated with small-vessel disease and lead to a higher risk of death, and poor cardiac and neurological outcome. However, several questions remain open and need further investigations. PMID- 10473978 TI - Multiple brain infarcts: clinical and neuroimaging patterns using diffusion weighted magnetic resonance. AB - The capability of diffusion-weighted (DW) magnetic resonance imaging (MRI) to identify very early ischemic brain injury better than conventional MRI is well known. This technique, which successfully discriminates acute from old infarcts, is particularly useful in patients with multiple brain infarcts (MBI). Among 142 patients with acute stroke consecutively admitted to our primary care center, we selected 43 patients with two or more brain infarcts on conventional MRI. All patients presented with clinical deficits consistent with acute cerebral ischemia and underwent conventional spin echo for T(1) (T1-WI) and T(2)-weighted images (T2-WI), T(1)-W gadolinium-enhanced images, and echo-planar technique for DW MRI sequences. Patients underwent DW MRI examinations within 15 days of stroke onset (mean +/- SD: 3 +/- 3 days). In all but 1 case, the infarcts detected on DW MRI were also visible on T2-WI. The different signal pattern on DW MRI, compared with T2-WI, facilitated the detection of acute infarcts in all patients. T1-WI with gadolinium enhancement was only helpful in 5 (11.6%) patients. DW MRI enabled precise clinicotopographic correlations in 79% of our patients and provided additional clinically relevant findings in 72% of the patients. Based on the neuroradiological findings, patients were divided into three clinicotopographic types of MBI as follows: 13 patients (30.2%) presented with multiple acute infarcts, 24 patients (55.8%) with a single acute infarct and multiple old infarcts, and 6 patients (13.9%) with multiple acute and old infarcts. In conclusion, DW MRI can easily be added to conventional MRI in order to be able to distinguish acute from old infarcts, and to identify acute multiple lesions. Therefore, a better correlation between clinical symptoms and the site of lesions can be obtained, considerably improving patient care. PMID- 10473979 TI - Indirect discharges as an early nerve conduction abnormality in the Guillain Barre syndrome. AB - Abnormal electrophysiological findings may be delayed in the Guillain-Barre syndrome (GBS). In this study, conducted in 29 GBS patients, 23 patients (group 1) fulfilled the electrodiagnostic criteria for GBS [1]at the time of initial investigation, performed on average 22.3 days after onset of symptoms; whereas the 6 patients (group 2) who did not fulfil the criteria were first seen earlier, on average after 9.7 days. During the initial nerve conduction studies, numerous late responses, distinct from F waves, were observed in 26 of the 29 patients (90%), that is in 20 patients of group 1, and in all patients of group 2. Most of these responses, if not all, were indirect discharges (iDs) caused by proximal re excitation on motor axons. If the iDs had been included in the electrodiagnostic criteria of GBS, all the patients of this series would have been diagnosed at the initial electrophysiological examination. We conclude that iDs occur frequently in GBS and are observed early in the disease. We recommend adding the presence of multiple iDs to the classical electrodiagnostic criteria of GBS. PMID- 10473980 TI - Verbal working memory deficit in Parkinson's disease subjects. AB - The aim of the present work was to evaluate verbal working memory in patients affected by Parkinson's disease (PD). In particular, tests that explore the functionality of the Articulatory Loop and of the Central Executive during verbal tasks were given. PD patients displayed normal phonological similarity, word length and word frequency effects in a Word span task, thus demonstrating adequate retention capacity of the phonological store, normal functioning of the articulatory rehearsal mechanism and a normal contribution of lexical-semantic knowledge to verbal immediate recall. In the Brown-Peterson task, PD patients showed abnormal performance decay on the letter recall task when articulatory rehearsal was inhibited by a serial subtraction concurrent task. These data provide evidence for normal functioning of the Articulatory Loop in PD patients. However, when the verbal recall task is more attention demanding, PD patients show deficient performance levels, presumably due to depleted processing resources by the Central Executive. PMID- 10473981 TI - An extensive search for autoantibodies to myelin basic protein in cerebrospinal fluid of non-multiple-sclerosis patients: implications for the pathogenesis of multiple sclerosis. AB - Inflammation of multiple sclerosis (MS) brain and spinal cord tissue consists of macrophages, T lymphocytes and cytokines as well as B lymphocytes and immunoglobulins (IgGs). IgG can be detected in high concentrations in both central nervous system tissue and cerebrospinal fluid (CSF). Using a sensitive radioimmunoassay (RIA), autoantibodies to myelin basic protein (anti-MBP) can be detected in the CSF of 90-95% of MS patients with active disease. The purpose of the present report was to determine whether these same autoantibodies can be reliably detected in non-MS patients. Between 1978 and 1998, CSF was collected from 1,968 control non-MS patients with psychiatric, inflammatory and noninflammatory neurological diseases as well as nonneurological systemic diseases, and anti-MBP were measured by the same RIA used to detect anti-MBP in MS CSF. Anti-MBP were undetectable in 98% of CSF samples from non-MS controls. In the remaining 2% of control samples, CSF IgGs capable of binding to MBP in vitro were unpredictably detected. This latter group included 1% of patients with miscellaneous diseases such as encephalomyelitis, 5 siblings with familial spastic paraparesis, rare patients with strokes, Wernicke-Korsakoff's syndrome, inherited leukodystrophy, motor neuron disease and some patients with miscellaneous spinal cord diseases. An additional 1% of patients included a group with neurological symptoms suggestive of early or predisseminated MS. The high prevalence of free and/or bound anti-MBP in the CSF of MS patients and the rare and unpredictable occurrence in the CSF of non-MS patients suggest that autoimmunity to MBP may be operative in the demyelination of MS. Molecular clones of anti-MBP with specificity towards variable surface or cryptic MBP epitopes in vivo may determine whether or not they are involved in the demyelinating process, and this variability may also be present within the MS population. Potential mechanisms of anti-MBP-mediated demyelination in MS patients are discussed. PMID- 10473982 TI - Practice variability in management of transient ischemic attacks. AB - To define practice patterns in the management of transient ischemic attacks (TIAs), we surveyed practicing neurologists attending an educational conference in San Francisco, evaluating management decisions in 2 TIA case vignettes. In a vignette describing a hemispheric TIA 1 day prior with ipsilateral bruit, 53% chose admission, 47% elected an outpatient work-up, 28% treated with intravenous heparin and 70% chose aspirin, reflecting the disagreement about medical management of carotid stenosis in the literature. There was more agreement in the second case, a posterior circulation TIA 1 day prior with atrial fibrillation, in which 84% chose hospital admission, 74% chose intravenous heparin and 90% treated with some form of anticoagulation. There are areas of important practice variability in the management of TIAs. Further research is justified to guide patient care decisions in TIA patients. PMID- 10473983 TI - Classical Friedreich's ataxia and its genotype. AB - Fourteen patients with classical features of Friedreich's ataxia (FRDA) were examined. The clinical diagnosis of FRDA was afterwards confirmed in all patients by the appropriate DNA investigation which showed markedly increased amounts of GAA repeats on both alleles of the frataxin gene. None of our patients presented with atypical features such as late-onset FRDA, FRDA with retained deep tendon reflexes or with a very slow course. Five of them are not yet confined to a wheelchair. But for 1 patient who died at age 36 years and had the largest number of GAA repeats on both alleles, there was no significant correlation between number of repeats in the shortest allele, age at onset, age at wheelchair dependence, duration of the disease and main clinical signs. All patients but 3 had between 500 and 1,050 GAA repeats. The 3 patients with, respectively, 400, 450 and 500 repeats on the shortest allele had a clinical course comparable to the other patients. Even in the case of variations in the number of repeats in the same sibship, there were only modest differences between the siblings concerning age at onset of the disease, symptoms and signs and age at wheelchair dependence. There were no qualitative differences in the main clinical features and laboratory investigations in the full-blown phase of the disorder. Molecular biology has become a major element in the diagnosis of FRDA. DNA testing for FRDA should be applied to every case of idiopathic autosomal recessive or sporadic ataxia. However, the clinical features of FRDA remain fully characteristic in many patients and keep their diagnostic value. PMID- 10473984 TI - Intravesical instillation of capsaicin in urology: A review of the literature. AB - OBJECTIVES: Interest in the intravesical instillation of capsaicin is growing among urologists. Its efficacy on detrusor hyperreflexia, hypersensitive bladder disorders and bladder pain has been reported in several studies. However, the lack of common evaluation parameters and the absence of consensus concerning a protocol of instillation hamper the interpretation of results. The purpose of this review is to better delineate the indications and optimum protocol for intravesical use of capsaicin. METHODS: Eight open and two placebo-controlled human clinical trials were analyzed. All 200 patients involved had lower urinary tract disorders. RESULTS: Clinical or urodynamic symptoms improved in 84.3% of the patients who received intravesical capsaicin for neurogenic hyperreflexic bladder, a significantly greater efficacy than that of placebo. Capsaicin may also be beneficial in patients who have non-neurogenic disorders. Whether or not the patients has a neurologic deficit, side effects appear during and in the period immediately following instillation. CONCLUSIONS: Intravesical capsaicin appears to be indicated in neurogenic hyperreflexic bladder, but is less effective against detrusor instability, hypersensitive bladder disorders or pelvic pain. The best instillation protocol and long-term tolerance remain to be established. PMID- 10473985 TI - Detection of prostate-specific antigen- or prostate-specific membrane antigen positive circulating cells in prostatic cancer patients: clinical implications. AB - OBJECTIVES: To evaluate the clinical benefit from using circulating prostate specific antigen (PSA) and prostate-specific membrane antigen (PSM) mRNA detection in prostate cancer staging and in follow-up. METHODS: Nested reverse transcriptase-polymerase chain reaction (RT-PCR) assays were performed on RNA extracted from blood drawn from 56 patients with prostate cancer before any treatment. Additionally, assays were done on posttreatment samples from 50 patients who were followed up by serum PSA level, to determine whether any relationship exists between RT-PCR results and tumor recurrence. The prostate cell specificity of assays was evaluated by analysis of 21 blood samples from women or cystoprostatectomized men. RESULTS: With PSM RT-PCR assay, good sensitivity and prostate cell specificity could not be attained together, since high PSM mRNA illegitimate expression has been shown in some healthy donor bloods. For this reason, only PSA RT-PCR assay was used as a clinical marker. PSA mRNA was detected in peripheral blood of 4 out of 31 patients with clinically localized prostate cancer. It showed no relationship to the pathologic stage, but significant relationship to metastatic status, lymph node involvement and Gleason score. During follow-up, circulating PSA mRNA was detected in 8 out of 17 (47%) patients in treatment failure and in only 1 out of 33 (3%) successfully treated patients, with significant relationship between RT-PCR results and concomitant serum PSA levels. CONCLUSION: Our study reveals no significant advantage to PSA RT-PCR assay (1) in improving the staging of clinically localized prostate cancer or (2) in follow-up treatment failure, as compared to the usual recurrence marker (serum PSA). Additional investigations are needed to determine the ultimate significance and the management of patients with positive PSA RT-PCR assays. PMID- 10473986 TI - Molecular heterogeneity of free PSA in sera of patients with benign and malignant prostate tumors. AB - OBJECTIVE: To analyze free prostate-specific antigen (f-PSA) in sera from patients with prostate cancer (PCa) and benign prostatic hyperplasia (BPH), and to detect possible differences in subtypes as potential diagnostic parameters. MATERIALS AND METHODS: PSA was purified from sera by an immunoaffinity procedure developed on the basis of oriented antibody immobilization, and subjected to size exclusion chromatography (SEC), Western blotting, and N-terminal amino acid sequencing. RESULTS: The novel procedure allowed the purification of PSA with high yield from sera containing PSA <10 ng/ml. SEC under nonreducing conditions as well as Western blots demonstrated the presence of several molecular forms of f-PSA. Three of the smaller polypeptides exhibited the N-terminal sequence of PSA while one represented the C-terminal fragment Lys(146)-Pro(237). Shortening of some polypeptides by the N-terminal amino acid Ile(1) suggestive of aminopeptidase action was also observed. No propeptide sequence could be detected, and none of the bands from patient sera reacted with antibodies raised against propeptide antigens. BPH sera expressed higher proportions of smaller PSA fragments per unit p33, and contained significant amounts of fragments <14,000 which appeared to be very low or absent from most PCa sera. CONCLUSIONS: f-PSA as obtained from BPH and PCa sera represents a heterogeneous fraction. The major component (p33) is not in the nicked form and does not contain proPSA. Diagnostic potential could arise from the quantitative differences of the smaller PSA derivatives seen between PCa and BPH sera. PMID- 10473987 TI - Antiandrogen hepatotoxicity in patients with chronic viral hepatitis. AB - OBJECTIVE: To assess whether patients with chronic viral hepatitis are at an increased risk for antiandrogen hepatotoxicity. METHODS: We retrospectively reviewed 121 prostate cancer patients who received long-term antiandrogen, either flutamide (n = 56) or cyproterone acetate (n = 65), and had normal pretreatment serum alanine aminotransferase (ALT) levels. Serological markers of hepatitis B and C viruses (HBV and HCV) were checked in 42 of the 121 patients. RESULTS: Twenty-two (18%) of the 121 patients had ALT elevations during antiandrogen therapy. Thirteen (59%) of the 22 patients were positive for either one of the two viral markers, including 7 for HBV, 4 for HCV, and 2 for both. This percentage was higher than the combined prevalence rate of positivity for HBV and/or HCV markers (<20%) in Taiwan. There was no significant differences in the percentage of positive makers among the two antiandrogen groups (p = 0.092). Although a higher incidence of hepatotoxicity was noted in the flutamide (13/56, 23%) than in the cyproterone acetate group (9/65, 14%), there were no significant differences between the two groups (p = 0.27). The time period between initiation of antiandrogen and first ALT elevation varied significantly (from 4 to 1,398 days with a median of 151 days). Half of the 14 HBV carriers and all of the 6 patients with anti-HCV developed antiandrogen hepatotoxicity. CONCLUSION: Our limited data suggested that patients with chronic viral hepatitis probably are at a higher risk of developing antiandrogen hepatotoxicity. Close monitoring of liver functions in patients with chronic viral hepatitis is advised if antiandrogen therapy is necessary. However, a large-scale study is necessary for a definitive conclusion. PMID- 10473988 TI - Mortality and morbidity after nephrectomy for renal cell carcinoma using a transperitoneal anterior subcostal incision. AB - OBJECTIVES: We have reviewed our surgical experience to document intra- and postoperative mortality and morbidity in 656 patients with renal cell carcinoma who underwent nephrectomy through a transperitoneal anterior subcostal incision (TASI). MATERIALS AND METHODS: From 1986 to 1997 we performed 656 nephrectomies for renal cell carcinoma using a TASI. Details of the surgical procedure are presented together with a retrospective analysis of the postoperative data concerning both the patient and the complications related to this approach. RESULTS: The mean time of operation was 130 min and the mean discharge from hospital 11 days. An additional surgical procedure in relation with the cancer facilitated by this approach was necessary in 2.1% of cases. The rates of intra- and postoperative complications were respectively 6.4 and 29.7%. The rate of intestinal complications was 1.8% and a splenic injury occurred in 8% of left nephrectomy. The mortality rate was 0.6%. CONCLUSIONS: The TASI is a large convenient incision which allows safe control of the renal pedicle in a very large number of renal tumors, even those located in the upper pole of the kidney. The rate of gut complications is very acceptable. Splenic injury is the major problem during left nephrectomy but careful dissection and surgical experience could decrease this complication, especially in case of upper pole renal tumor. We consider the TASI to be the main radical nephrectomy incision for renal cell carcinoma. PMID- 10473989 TI - Radical nephrectomy for renal cell carcinoma: Is adrenalectomy necessary? AB - OBJECTIVES: The role of simultaneous adrenalectomy in combination with radical nephrectomy in the treatment for renal cell carcinoma (RCC) remains controversial. With nephron-sparing surgery being commonly applied, the indication for adrenalectomy has to be critically assessed. PATIENTS AND METHODS: In a retrospective analysis the outcome of 589 patients, who underwent ipsilateral adrenalectomy along with radical nephrectomy in the treatment for RCC between 1985 and 1997 at our institution, was evaluated. The mean follow-up time was 34 months (range 1-95). RESULTS: Histologically an ipsilateral adrenal metastasis was found in 19/589 patients (3.2%). 16/19 patients had >/= T3, 3/19 had T1 tumours. The average size of the primary tumours with adrenal metastasis was 7.8 cm (range 2.3-13) in diameter with no preferential primary tumour site within the kidney (6/19 upper, 4/19 middle and 9/19 lower third). Only 4/19 patients had suspect adrenal findings in preoperative diagnostics (ultrasound, CT scan). 6/19 (31.5%) patients with adrenal metastasis are alive without evidence of disease at a mean of 41 months (range 11-95) after surgery for RCC. CONCLUSIONS: The probability of adrenal metastasis correlates with primary tumour stage, but not with its location within the kidney. The preoperative diagnostics are not reliable concerning small adrenal metastases. We thus still recommend simultaneous adrenalectomy in those cases where radical nephrectomy in patients with RCC is indicated. PMID- 10473990 TI - Late recurrence and progression after a long tumor-free period in primary Ta and T1 bladder cancer. AB - OBJECTIVE: To determine the probability and risk factors of recurrence and progression (to T2 or worse) after a long tumor-free period in patients with superficial (stage Ta and T1) bladder cancer. PATIENTS AND METHODS: A consecutive series of 100 patients with superficial bladder cancer who remained tumor-free for longer than 4 years after initial treatment were reviewed. The rates of recurrence and progression were statistically assessed and the significance of risk factors was determined. RESULTS: Of the 100 patients, 24 (24. 0%) recurred within 15 years after the initial treatment. The 10- and 15-year recurrence-free rates were 76.0 and 59.6%, respectively. Among the clinicopathological variables examined, intravesical chemotherapy was determined by a log-rank test to be a significant unfavorable risk factor for late recurrence (p < 0.001). Progression of the tumor occurred in 5 patients. Four variables including presence of multiple tumors, involvement of the bladder neck, positive urine cytology, and intravesical chemotherapy were found by a univariate analysis to be significant risk factors for late progression (p < 0.05). Among these factors, initial presence of multiple tumors (3 or more) was determined by a multivariate analysis to be an independent risk factor for late progression. CONCLUSION: Recurrence and progression continue to occur in patients with superficial bladder cancer even after long periods of dormancy. Regular follow-up urological assessments should be continued until at least 15 years of tumor-free existence, especially in patients treated by intravesical chemotherapy or those initially having multiple tumors. PMID- 10473991 TI - Evaluation of DNA ploidy combined with a cytometric proliferation index of imprints from core needle biopsies in prostate cancer. AB - OBJECTIVE: To evaluate if DNA ploidy analysis with a proliferation index (PI) derived from DNA cytometry of imprints from core needle biopsies predicts disease progression in patients with prostate cancer. METHODS: Touch imprints were done on a consecutive series of core needle biopsies taken by the same urologist from 240 patients with suspected prostate cancer, 137 (46%) of whom were found to have prostate cancer and included in the study. Scattered cells to the right of the image cytometry (ICM) ploidy-establishing peak, the S-phase fraction, and those in the G2M area of the ICM DNA histograms, were counted in percent of the total number of tumor cells, this value being designated the ICM PI. Based on previous results in archival fine needle aspiration material, the following classification was used: DNA group I, diploid tumor with a low PI; DNA group II, diploid tumor with an intermediate PI and tetraploid tumor with a low or intermediate PI, and DNA group III, diploid or tetraploid tumor with high PI and all tumors with an aneuploid pattern. RESULTS: Correlation was found to exist between DNA groups I III and Gleason score (GS) (p < 0.0001), T-stage (p = 0.006), M-stage (p = 0.009) and disease progression (p < 0.0001). Among the 39 patients who had curative treatment and GS 5-7, the progression-free survival rate was 100% in DNA group I, as compared with only 38% in DNA group II and 55% in DNA group III within the follow-up period (p = 0.008). CONCLUSION: DNA ploidy combined with a PI derived from image cytometry of imprints from core needle biopsies yields additional prognostic information in patients with GS 5-7. Diploid tumors with a low PI (DNA group I) are associated with a low risk of disease progression. PMID- 10473992 TI - Value of power Doppler sonography in the investigation of erectile dysfunction. AB - OBJECTIVES: The poor sensitivity of conventional color-coded Doppler sonography (CCD) for low-flow signals limits its use for investigating patients with erectile dysfunction. Power Doppler sonography (PD) has recently been described for enhanced visualization of the microcirculation. Aim of this study was to determine the value of PD to demonstrate penile vascular pathophysiology as compared with conventional techniques. METHODS: 33 consecutive men with erectile dysfunction were investigated using the standard workup with conventional CCD and cavernosography before and after prostaglandin E(1) intracavernosal injection. Patients were subdivided into an arteriogenic, a venogenic, or a psychogenic group according to findings in the standard diagnostic workup. PD was used in addition to the standard protocol to demonstrate microcirculation, arterial blood flow, and venous leakage. The accuracy of the diagnosis obtained by PD and response to intracavernosal injection was compared with the clinical outcome in these groups at 6 months. RESULTS: PD was found to be superior to CCD in visualizing cavernosal microcirculation. In addition, arterial flow at basal peak systolic velocity was demonstrated in all patients with PD, whereas a signal sufficient for evaluation was obtained with CCD in only 69.7% (23/ 33). No significant difference in the maximal peak systolic velocity was noted using either PD or CCD. The positive predictive value of PD for venous leakage was poor (60%) when compared with cavernosography. PD used in conjunction with the response to intracavernosal injection was found to reliably predict the clinical outcome in the arteriogenic (p = 0.0007), the venogenic (p = 0.005), and the psychogenic group (p = 0.0002). CONCLUSIONS: Our data indicate that PD improves the evaluation of penile microcirculation and arterial function, but fails to reliably demonstrate venous leakage alone. Nevertheless, in most patients cavernosography could have been avoided by the aid of PD, since the underlying pathology can be calculated at a high predictive value without the need of further invasive tests. Therefore, with the aid of PD, the morbidity for patients being investigated for erectile dysfunction can be significantly reduced. PMID- 10473993 TI - Our experience on the association of a new physical and medical therapy in patients suffering from induratio penis plastica. AB - OBJECTIVES: To check the efficiency of shock waves in the treatment of induratio penis plastica. The Minilith SL1, successfully used in orthopedic or salivary stones because of its lithotriptic power, can be used to break plaques in Peyronie's disease. METHODS: A total of 130 patients affected with Peyronie's disease were entered into a prospective trial. Patients with completely calcified plaques as determined by ultrasound evaluation were excluded. We divided the patients into three treatment groups: (A) shock waves alone in 21 patients; (B) a combination of shock waves and verapamil (perilesional injection) in 36 patients, and (C) verapamil alone in 73 patients. First, we treated all groups A and B patients 3 times, 20 min each time, with a Minilith SL1, and then only the patients of the second group received a complete cycle of twelve injections of verapamil (10 mg) every 2 weeks for 6 months. The group of 73 patients (group C) treated during the previous 2 years with a medical therapy (only injection of verapamil) was used as a control group. RESULTS: Ultrasound evaluation showed a reduction of plaque in 11/21 group A patients and 7/36 group B patients. The treatment was tolerated very well and only 11 petechiae in some patients were noticed after ESW treatment. CONCLUSIONS: The therapeutic association of shock waves with verapamil injection is an effective nonoperative treatment for the stabilization of Peyronie's disease. PMID- 10473994 TI - TURP with the new superpulsed radiofrequency energy: More than a gold standard. AB - OBJECTIVE: Superpulsed radiofrequency, a new energy, has been developed and applied in transurethral resection of the prostate (TURP) using a regular cutting loop. The aim of the study was to evaluate the clinical application of superpulsed radiofrequency to reduce bleeding morbidity and to improve patient outcome. The original results of the experimental and clinical study are presented. METHODS: From November 1997 to June 1998, 37 patients were submitted to a clinical, nonrandomized study. The age range was 45-81 years. All patients completed the International Prostate Symptom Score (IPSS) and were examined using digital rectal examination, prostate-specific antigen level, transrectal ultrasound, free uroflow, and urodynamic pressure flow study. Hemoglobin was tested before and after surgery. Follow-up was at 3 and 6 months. All operations were performed with a short-acting spinal anesthesia and the resected prostate weight was 18-66 g. RESULTS: 36 patients were treated as 1-day surgery; 1 patient was hospitalized for 1 day. Intraoperative bleeding was very low (hemoglobin tested before and after surgery): the postoperative Hb was <1 g/dl in 34 patients, and Hb = 2 g/dl in 3 patients. Postoperative bleeding was minimal; late bleedings were not observed. Stress incontinence or incontinence were not observed. At 6 months, the patients were asked about any changes in their sexual lives, and no symptoms of impotence were reported. The peak flow rate (PFR) ranged from 4 to 7 ml/s at baseline, with a medium PFR of 5.5 ml/s. At 3 and 6 months the average increase in PFR was 11 and 10 ml/s, respectively, in all patients. CONCLUSIONS: As shown by this study, superpulsed radiofrequency is a safe and effective technology. It is superior to high-frequency surgical units and high-frequency coagulating intermittent cutting in terms of whole intraoperative bleeding (hemoglobin tested before and after surgery) and for the impressive impact on minimizing postoperative bleeding. In terms of improvement in urinary flow, the results of TURP with superpulsed radiofrequency at 3 and 6 months are comparable with the average PFR generally reported for TURP performed with high-frequency surgical units. Moreover, superpulsed radiofrequency TURP enhances the gold standard and makes TURP competitive with the other alternative treatments available for benign prostatic hyperplasia. Further studies could better define and extend the application of this new form of energy either in urologic or general surgery. PMID- 10473995 TI - Tamsulosin 0.4 mg once daily: effect on sexual function in patients with lower urinary tract symptoms suggestive of benign prostatic obstruction. AB - OBJECTIVE: To evaluate the effect of tamsulosin, 0.4 mg once daily, on sexual function in comparison with placebo and alfuzosin, 2.5 mg three times daily, in patients with lower urinary tract symptoms (LUTS) suggestive of benign prostatic obstruction (BPO). METHODS: Data from 830 patients randomized into three European multicenter studies with similar protocols were analyzed. In two studies, patients were randomized to receive either tamsulosin, 0.4 mg once daily, or placebo, and in the third, patients were randomized to receive either a fixed dose of tamsulosin, 0.4 mg once daily, or alfuzosin, titrated to 2.5 mg three times daily. The studies employed a 2-week placebo run-in period, followed by a 12-week study period. Sexual function was assessed by related adverse events and by a sexual function score determined from a life-style questionnaire. RESULTS: Abnormal ejaculation occurred significantly more frequently in patients treated with tamsulosin than in those receiving placebo (p = 0.045); however, the incidence of abnormal ejaculation was similar in patients receiving tamsulosin or alfuzosin in the comparative study. Abnormal ejaculation was not perceived as a major problem by the patients since it resulted in few treatment discontinuations (n = 3). It was also reversible on drug withdrawal. There was no difference between tamsulosin and placebo or alfuzosin with regard to the occurrence of decreased libido or impotence. In addition, there was no significant difference in the change in sexual function score between patients treated with tamsulosin and those treated with alfuzosin. Compared with patients receiving placebo, there was, however, a significant improvement in total sexual function score in patients receiving tamsulosin (p = 0.042). CONCLUSIONS: Tamsulosin, 0.4 mg once daily, is well tolerated and has no overall negative impact on sexual function compared with placebo or alfuzosin. Compared with placebo, tamsulosin may even improve sexual function. PMID- 10473996 TI - A review of ninety-two obstructive megaureters in children. AB - OBJECTIVES: The evolution of clinical presentation, age of surgery and therapeutic approach of obstructive nonrefluxing megaureters (OMU) in children throughout the years has been retrospectively evaluated. METHODS: 78 children with 92 stenotic ureterovesical junctions (UVJ) were reviewed. 66 underwent surgery at a median age of 20 months, after a median of 10.5 months of conservative treatment with prophylactic antibiotics. 21 OMU were diagnosed prenatally, 71 because of symptoms at later age. 15 ureters (12 children) (24% in the prenatal, 14% in the second group) were treated in a conservative way for 2 years with antibiotics. In the prenatal group 33% needed a reimplantation with tailoring and 10% without tailoring while in the other group the figures are reversed: 39% without and 21% with tailoring. 28% in the prenatal group and 17% of the second group were reimplanted at a mean of 15 months after a primary cutaneous ureterostomy. Three of 5 ureteroceles were treated by endoscopic incision; 4 had an immediate nephroureterectomy. The mean follow-up is >70 months. RESULTS: By prenatal diagnosis the number of conservatively treated cases increased from 14 to 24%. Indications for surgery remained unchanged: recurrent infection and poor kidney function. Both approaches resulted in stabilization of pretreatment renal function; nearly half of the DMSA scans showed a R:L difference of >20% at follow-up. Ureterostomy for infected deteriorating kidneys rapidly ameliorated the function and resulted in shrinking of the ureteral diameter making tailoring at reimplantation unnecessary. One of the 3 endoscopically incised ureteroceles required later reintervention. CONCLUSIONS: Male:female (3:1), left:right (2:1) prevalence and high associated urological (30%) and nonurological (19%) pathology is found. Unsatisfactory reliability of tests for obstruction diagnosis and a referral bias explains the larger number of conservatively treated ureters in the prenatal group. Despite prenatal diagnosis, the age for surgery was not altered since the indications remained identical. Most OMU can be treated by a simple or tailored reimplantation of the ureter after resection of the stenotic segment. A temporary ureterostomy in small children with refractory infections restores function and avoids the necessity for tailoring at final reconstruction. One of 3 endoscopically incised ureteroceles needed surgery at a later stage. DMSA shows stable function after reimplantation. PMID- 10473997 TI - Nephrogenic adenoma: A rare bladder tumor in children. AB - OBJECTIVE: Nephrogenic adenomas of the urinary bladder are rare benign tumors in children. The purpose of our study was to obtain information about the sex distribution, presenting symptoms, intravesical locations, therapy and recurrence rates in pediatric nephrogenic adenomas. PATIENTS AND METHODS: The records of 3 children with nephrogenic adenoma of the urinary bladder diagnosed between 1990 and 1997 were reviewed to evaluate the initial symptomatology, diagnostic examinations and findings, therapeutic procedures and clinical outcome and recurrence rates. Furthermore our data are compared to the findings of all children reported in the literature. RESULTS: Including the 3 cases reported by us, the data on 18 children with nephrogenic adenoma of the bladder could be analyzed. There was a significant predominance of girls compared to boys (5:1); the medical history in all cases was remarkable for previous bladder surgery 3 months to 7 years prior to tumor diagnosis. Most children presented with unspecific symptoms of gross hematuria, dysuria and bladder instability and in all cases the final diagnosis was established after cystoscopy and histopathologic review of a tumor biopsy specimen. Therapy consisted of transurethral resection in 15 cases, partial cystectomy and open excision in 2 and 1 case, respectively. Tumor recurrence developed in 80% of the children with a latency period of 4 years. CONCLUSIONS: Nephrogenic adenomas represent an epithelial response of the urothelium to chronic inflammation or previous trauma resulting in urothelial metaplasia and the development of papillary lesions. Current treatment of choice consists of transurethral resection and fulguration of the base of the tumor and periodic cystoscopy. PMID- 10473998 TI - Effects of acute urinary bladder overdistension on bladder response during sacral neurostimulation. AB - OBJECTIVE: Urinary retention and micturition disorders after overdistension are clinically well-known complications of subvesical obstruction. We attempted to evaluate whether bladder overdistension influences bladder response and whether overdistension supports detrusor decompensation. METHODS: Following lumbal laminectomy in 9 male foxhounds, the sacral anterior roots S2 and S3 were placed into a modified Brindley electrode for reproducible and controlled detrusor activation. The bladder was filled in stages of 50 ml from 0 to 700 ml, corresponding to an overdistension. At each volume, the bladder response during sacral anterior root stimulation was registered. After overdistension, the bladder was refilled stepwise from 0 to 300 ml and stimulated. RESULTS: In all dogs, the bladder response was influenced by the intravesical volume. The maximum pressure (mean 69.1 cm H(2)O) was observed at mean volume of 100 ml. During overdistension, a significant reduction in bladder response of more than 80% was seen. After overdistension, a significant reduction in intravesical pressure of 19.0% was observed. In 2 cases, reduction in bladder response was more than 50% after a single overdistension. CONCLUSIONS: We conclude that motoric bladder function is influenced during and after overdistension. A single bladder overdistension can support acute and long-lasting detrusor decompensation. In order to protect motoric bladder function, bladder overdistension must be prevented. PMID- 10473999 TI - Dyslexia: oral and written language disorder. A new look at old links. AB - It is now generally accepted that written language attainment is directly associated with oral language ability, and that deficits in oral language will be reflected in written language. This paper examines the links that exist between these two modes of communication from a historical perspective, as well as from current research. The concept and terminology of dyslexia - written language disability - are explored. The impact of deficits in phonology, vocabulary, semantics and syntax on the acquisition of written language is discussed. PMID- 10474000 TI - Tracheoesophageal voice restoration: origin--evolution--state-of-the-art. AB - Tracheoesophageal voice has gradually evolved worldwide as a viable option for voice restoration following total laryngectomy. This article presents a brief overview of the principal surgical and prosthetic aspects of this technique. PMID- 10474001 TI - Managing aphasia: then and now. AB - The nature and attitudes of the professionals who 'manage' aphasia have changed significantly over the 75 years the IALP has existed. This review shows how IALP publications have reflected this, and considers recent developments, particularly in respect of the role played by the cognitive neuropsychological approach in evidence-based healthcare and by conversational approaches in the empowerment of aphasic people. PMID- 10474002 TI - Vocal fold vibration and voice quality. AB - There have been various methods proposed for diagnosing laryngeal pathology by analyzing the pathological voice. However, even if two diagnoses are the same, the voice quality might not be the same. In other words, it is difficult to diagnose laryngeal pathology only by acoustic parameters. In this study, we analyze the vibration patterns seen in various pathological cases to explain pathological voice quality. PMID- 10474003 TI - Children of yesterday, today and tomorrow: global implications for child language. AB - The entire global population is waiting for new opportunities of the new millennium. This paper will report selected global trends; describe briefly children of yesterday, today and tomorrow; detail a case study explaining the multiple challenges of working with children in a multicultural environment and probe into the implications for the future of child language. PMID- 10474004 TI - Task-based profiles of the dysarthrias. AB - The dysarthrias are associated with a variety of motor disturbances distributed over several motor systems of speech production. The features of a given dysarthria often vary with the speaking task, and this task-dependency affords insights into the responsible neural lesion and its effects on the motor regulation of speech. Each task also is amenable to quantitative analyses with acoustic or physiologic methods, and these analyses may redefine the value of these speaking tasks. This article considers task-based analyses for the dysarthrias associated with Parkinson's disease, cerebellar disease, and stroke. PMID- 10474005 TI - Generalizable outcomes of bilingual aphasia research. AB - A number of constructs developed to account for bilingual aphasia phenomena have been advantageously extended to increase our understanding of language representation, processing, breakdown and rehabilitation in unilinguals as well. In particular, focus on the right-hemisphere-based pragmatic component of verbal communicative competence, the activation threshold, the control of resources, the role of emotion in second language acquisition and that of procedural vs. declarative memory, has led to the suggestion that unilinguals are in fact at one end of a continuum, with multilinguals who speak genetically unrelated languages at the other end. No function is available to the bilingual speaker that is not already available to the unilingual, unidialectal speaker. The only difference seems to be the degree of use the speaker makes of each of the relevant cerebral systems. PMID- 10474006 TI - Comprehensive rehabilitation after total laryngectomy is more than voice alone. AB - Comprehensive rehabilitation after total laryngectomy is more than voice alone. The removal of the larynx and the subsequent disconnection of the upper and lower airways not only has consequences for the vocal function but also for the respiratory system, and for the olfactory acuity of the patient. The results of the research program on these three subjects in the Netherlands Cancer Institute over the last 12 years are discussed in detail along with some of the relevant other literature on the topics. The considerable progress in these areas over the last 2 decades has clearly improved the quality of life of laryngectomized patients. PMID- 10474007 TI - Developmental language impairments with complex origins: learning from twins and multiple birth children. AB - Various factors that make language development vulnerable in twins and multiple birth children are discussed. Researchers have investigated single explanations for language impairment in twins. However, this paper argues that in individual cases the language impairment is more often the outcome of a complex interaction of factors. Some factors play a greater part than others in some cases. It is suggested that clinicians can develop a dynamic model of language acquisition from understanding the effect of multiple birth and other exceptional circumstances on language acquisition. This model is needed to underpin investigations into individual cases of language impairment. An approach to assessment is described that identifies factors operating in individual cases of multiple birth, providing an essential precursor to implementing appropriate case based intervention. PMID- 10474008 TI - Audiology on the way into the next millennium. AB - The article gives an overview of current developments in audiology. Present challenges and questions are discussed and possible solutions are suggested. The scope includes diagnostic and rehabilitative features of audiology and focuses on basic assessment of hearing functions, localization of the site of lesion, objective assessment of auditory functions, and hearing instruments, both traditional and implantable ones. Finally, new attempts at tinnitus treatment and possible approaches to the future prevention of hearing loss and protection of hearing by medication are briefly described. Due to the vast diversity of modern audiology only the major trends are covered. Thus this article does not claim to provide a complete survey of all that is going on in audiology today. PMID- 10474009 TI - Voice research and clinical needs. AB - The purpose of this paper is to provide a review of recent advances in voice research with special focus on clinical aspects and on the speaking voice. First an update on histological findings concerning the multilayer structure and the biomechanics of the vocal folds is given. Other topics are vocal fold closure patterns, the development of the voice from childhood to senescence, and theoretical aspects of vocal function from an acoustic-aerodynamic point of view. Special emphasis is put on voice parameters, which can be assumed to be relevant to underlying physiology, and how they can be captured by different techniques. PMID- 10474010 TI - Recent developments in speech motor research into stuttering. AB - This paper discusses recent speech motor research into stuttering within the framework of a speech production model. There seems to be no support for the claim that stutterers differ from nonstutterers in assembling motor plans for speech. However, physiological data suggest that stutterers may at least have different ways of initiating and controlling speech movements. It is hypothesized that stuttering may be the result of a deficiency in speech motor skill. Furthermore, objections to the use of stuttering frequency as a severity index are formulated and future developments in the assessment of speech motor behavior in stuttering are described. PMID- 10474011 TI - Voice problems at work: A challenge for occupational safety and health arrangement. AB - In modern societies about one third of the labor force are working in professions in which voice is the primary tool. Voice problems are common in general, but they are even more common in professions in which there is heavy vocal loading, i.e. professions that do not only require prolonged voice use, but also involve extra loading factors such as background noise, long speaking distance, poor room acoustics, lack of adequate equipment like voice amplifiers etc. School and kindergarten teachers can be considered to represent professions with heavy vocal loading. The occupational safety and health arrangements of voice and speech professionals are poorly developed as compared to many other professions. However, the existing legislation could be used to support efforts to improve the working conditions of this large but heterogeneous group. PMID- 10474012 TI - Augmentative and alternative communication in logopedics. AB - Like logopedics, augmentative and alternative communication (AAC) has a short history and a long past. In the 1950s and 60s the field of AAC emerged as a response to the need of individuals who, despite years of 'traditional speech therapy', had not developed adequate oral communication skills, and for whom compensatory rather than remedial approaches were thus deemed advisable. Various trends like the shift from form to function in language development research, dissemination of sign language, use of non-speech graphic symbols, advances in computer technology, and international collaboration have all contributed to AAC's present-day status as a multidisciplinary enterprise involving speech language therapists and an array of other professions. Clinical experience, for lack of comprehensive empirical studies, seems to indicate that AAC is warranted for congenital as well as acquired communication disorders. PMID- 10474013 TI - The potential future of dysphagia: population-specific diagnosis and treatment. AB - Care of patients with dysphagia includes screening, diagnosis, treatment and management. This article examines the increasing evidence that to be most effective each of these aspects of patient care must be tailored to the patient's medical diagnosis, specific swallowing disorders and stage of recovery or degeneration. The need to define the optimal window of intervention for each patient type is also discussed. Future research needs are emphasized. PMID- 10474014 TI - IALP and the world: global perspectives. International Association of Logopedics and Phoniatrics. AB - IALP is an association that guards, coordinates, and enhances internationally the field of communicative sciences and disorders. On the occasion of its 75th anniversary an attempt is made in this presentation to introduce the Association and the lines of possible future development in relation to its global responsibilities. An introduction to human communication aims at highlighting the importance of the field of interest of IALP. The history and present status of IALP is given. Future development focuses on aspects of the specialty and on promotion of the goals of IALP with reference to the importance of developing international contacts. Finances, an important aspect in the development of any establishment, are referred to, stressing their sound features at present. Suggested global tasks are put forward as a visionary and even a missionary plan for IALP in its coming decades. PMID- 10474016 TI - Evidence for the synthesis of corticosteroid-binding globulin in human placenta. AB - We demonstrated the expression of corticosteroid-binding globulin (CBG) in human placenta using reverse transcription-polymerase chain reaction-Southern blot analysis and immunohistochemical and immunoblotting studies. In the RT-PCR Southern blot analysis, only one predicted PCR product was detected without nonspecific products in all samples of human placenta and 3A (tPA-30-1) human placental cells. In Western blot analysis, polyclonal anti-CBG antibodies recognized a protein of approximately 55 kD in the protein extracts prepared from 3A (tPA-30-1) cells. Additionally, CBG mRNA expression was demonstrated by in situ hybridization in the syncytiotrophoblasts. Immunohistochemical studies performed on the placenta demonstrated the presence of specific immunoreactivity in the syncytiotrophoblast layer surrounding the chorionic villi. These findings suggest that CBG is synthesized in human placenta during pregnancy in addition to its synthesis in the liver. PMID- 10474015 TI - Growth hormone treatment of children with Prader-Willi syndrome: effects on glucose and insulin homeostasis. Swedish National Growth Hormone Advisory Group. AB - Insulin and glucose homeostasis have been studied during growth hormone (GH) treatment in 19 prepubertal children with Prader-Willi syndrome (PWS) and compared with 11 healthy prepubertal obese children. Before treatment, insulin levels in children with PWS were lower (p < 0.01) than in healthy obese children. During GH treatment, fasting insulin levels increased in children with PWS (p < 0.001). Glucose levels were similar for PWS and obese children before treatment. Children with PWS showed a slow glucose disappearance rate (k = 1.7%) which deteriorated (k = 1.3%, p < 0.001) during GH treatment. HbA1c and fasting glucose levels remained normal. Thus, GH treatment of children with PWS resulted in increased insulin blood levels, unchanged fasting glucose and HbA1c but decreased glucose elimination rate after an intravenous glucose test. However, the observed dose-dependent increase in insulin levels during GH treatment, that reached supranormal concentrations in 6/19 patients, and the occurrence of NIDDM in 1 patient during follow-up suggest that close surveillance and low doses of GH should be applied, especially if the PWS patient is very obese. PMID- 10474017 TI - Use of the new US90 standards for TW-RUS skeletal maturity scores in youths from the Italian population. AB - Recently, 1997, Tanner and co-workers provided a new scale converting TW-RUS standard maturity scores to skeletal age for European North American youths (US90). The aim of the present study was to test if the accuracy of TW-RUS bone age assessments in the Italian population could be improved by evaluating the estimates obtained with this new scale in comparison with other standards (UK60: original British series, B70: Belgian series and S80: Spanish series). 1,831 hand wrist radiographs (Italian healthy subjects aged from 8 to 16.8 years) were evaluated. The US90 reference values are resulted the most suitable TW-RUS standards. Therefore, it seems useful to update the reference values of TW-RUS SMS in Italian youths, using this new scale. PMID- 10474018 TI - Increased urinary excretion of collagen crosslinks in type 1 diabetic children in the first 5 years of disease. AB - To analyze possible early abnormalities in bone resorption in type 1 diabetes mellitus the urinary excretion of the collagen crosslinks pyridinoline and deoxypyridinoline was evaluated by immunoassay in 26 prepubertal diabetic patients (mean age 7.8 +/- 1.6 years, mean duration 3.0 +/- 1.1 years) and 46 healthy children (age 8.3 +/- 1.3 years). Relationships with growth parameters (height-standard deviation score, body mass index and height velocity during the year preceding the study) and metabolic control were sought. Longitudinal and ponderal growth was normal in diabetic children. Urinary collagen crosslink excretion was 88.4 +/- 25 nmol/mmol creatinine (median 86, range 44-146) in diabetic patients and 65.6 +/- 19 nmol/mmol creatinine (median 61, range 32-108) in controls (p = 0. 0002). It was positively influenced by diabetic status (beta = 20.5) and negatively by age (beta = -6.41), controlling by sex and BMI (p = 0.0001). A positive correlation was found between collagen crosslinks and blood glucose (r = 0.48, p = 0.01) or HbA1c levels (r = 0.44, p = 0.02) evaluated at the time of the study, while no significant correlation was found with the mean HbA1c values assessed in the last year or throughout the whole duration of diabetes. Collagen crosslink excretion was significantly increased in patients who presented worsening of their metabolic control in the last 3 months. No relationship was found with the duration of disease or growth parameters. In conclusion, the elevated urinary excretion of collagen crosslinks in diabetic children indicates that bone resorption may be disturbed. Poor metabolic control influences the increased rate of bone resorption and may expose growing diabetic patients to a risk of bone loss. PMID- 10474019 TI - Effects of 1-year ipriflavone treatment on lumbar bone mineral density and bone metabolic markers in postmenopausal women with low bone mass. AB - In vitro studies have shown that ipriflavone affects both bone formation and bone resorption, but the effect in early-stage postmenopausal women with low bone mass and a high turnover of bone metabolism is unknown. In this prospective study, we randomly assigned 60 patients with postmenopausal osteopenia or osteoporosis to receive either 600 mg/day of ipriflavone or 0.8 g/day calcium lactate, and compared the effects on bone mineral density (BMD) from the 2nd to 4th lumbar vertebrae (L2-4) and bone metabolic markers before and after one year of treatment. In the iprifravone-treated (IP) group, L2-4 BMD was similar before and after treatment (0.78 and 0.77 g/cm(2), respectively), but in the calcium lactate treated (CL) group, L2-4 BMD decreased significantly from 0.81 to 0.79 g/cm(2) after 1 year of treatment (p < 0.0001). Furthermore, the rate of the decrease in L2-4 BMD was significantly greater in the CL group than in the IP group (p < 0.01). The median deoxypyridinoline (Dpd) level was significantly lower after 1 year of treatment (5.8 mmol/mmol creatinine [Cr]) than the baseline value (10.2 mmol/mmol Cr) in the IP, but not in the CL group, suggesting that IP treatment suppresses bone resorption. PMID- 10474020 TI - Near-normal linear growth in the setting of markedly reduced growth hormone and IGF-1. A case report. AB - A 14.2-year-old prepubertal boy diagnosed with complete-type growth hormone deficiency and tertiary hypothyroidism, keeps growing in the height range between -1 and -2 SD. He has been treated with levothyroxine only. To understand the growth mechanism of this boy, we analyzed the serum growth hormone (GH) with a radioimmunoassay (RIA), serum GH bioactivity with Nb2 and erythroid progenitor cell bioassays, and growth hormone-binding protein (GHBP) with a ligand-mediated immunofunctional assay (LIFA). In addition, IGF-1 and free IGF-1 were analyzed by immunoradiometric assay (IRMA) and insulin-like growth factor-binding protein-3 (IGFBP-3) by Western immunoblot. Peak GH-RIA responses to insulin, arginine and GH-releasing factor, and nocturnal GH secretion, were low (0.5-2.3 ng/ml); bioactive GH was low (0.313 ng/ml), and GHBP was elevated (84 ng/ml). The serum levels of IGF-1 and free IGF-1 were continuously low, 17.1-39.3 and 0.17-0.26 ng/ml, respectively. Moreover, serum IGFBP-3 levels were low (1.68- 1.39 mg/l) and IGFBP-3 protease activity was negative. Prolactin and insulin were in the normal range. The result of the assay for growth-promoting activity showed that the patient's serum stimulated normal erythroid progenitor cells twice as potently as did healthy thin adult control serum. These results suggest that GH and IGF-1 are not indispensable for maintaining physical growth in this boy. Thus, it appears that circulating GH and IGF-1 are not mandatory requirements for maintaining normal physical growth, and other, as yet uncharacterized, pathways or growth factors might be sufficiently compensatory under certain conditions. PMID- 10474021 TI - Effects of glycyrrhizin on production of vascular aldosterone and corticosterone. AB - This study is to confirm the role of glycyrrhizin on blood pressure and to test the effects of glycyrrhizin on production of vascular aldosterone and corticosterone in rats. Male Wistar rats received glycyrrhizin (Sigma) 200 mg/kg/day p.o. for 5 weeks, and blood pressure was monitored by a pressure transducer. Systolic blood pressure significantly increased in Wistar rats treated with glycyrrhizin compared to that without glycyrrhizin. Mesenteric artery perfusion ex vivo and pressor responses to norepinephrine were performed. The pressor responses to norepinephrine in mesenteric arteries treated with glycyrrhizin were significantly increased. The perfusate from the mesenteric arteries was collected and applied to a Sep-Pak C 18 cartridge column, used for reverse-phase high-performance liquid chromatography, and measured for both aldosterone and corticosterone by radioimmunoassay. Levels of aldosterone were decreased but those of corticosterone increased in perfusate from arteries treated with glycyrrhizin. RT-PCR showed that glycyrrhizin inhibited the expression of 11beta- HSD2 and CYP11B2 mRNA in mesenteric arteries. These results confirm that glycyrrhizin is able to induce hypertension, and provide evidence that it inhibits the transcriptions of both 11beta-HSD2 and CYP11B2 in the vasculature, leading to lower aldosterone and higher corticosterone production in vessels, and increased vasoconstrictor responses to norepinephrine. PMID- 10474022 TI - Hypoglycemic effects of insulin-like growth factor-1 in experimental uremia: can concomitant growth hormone administration prevent this effect? AB - The risk of hypoglycemia limits the clinical application of insulin-like growth factor-1 (IGF-1). Our studies aimed to evaluate the mode of occurrence as well as the prevention of this side effect. Acute administration (i.v. infusion) of IGF-1 in subtotal nephrectomized uremic (U), sham-operated ad libitum fed control (C) and sham-operated pair-fed control (P) rats led to hypoglycemia, though more expressed in P. Serum glucose levels decreased within 60 min after the IGF-1 administration by 40% in U, by 45% in C and by 52% in P (p < 0.05, U vs. P). Chronic administration (7 days) of 1, 4 and 8 mg/kg/day IGF-1 in U rats led to hypoglycemia in an increasing manner as the dose of IGF-1 increased. On the first day, 2 h after injection, serum glucose levels were 116.5 +/- 8.6, 110.4 +/- 12.4, 60,3 +/- 19.2 and 50.6 +/- 18.3 mg/dl, respectively (p < 0. 01). One week later, IGF-1 therapy proved to be less hypoglycemic in all the groups. On day 7, 2 h after injection the serum glucose levels were 118.9 +/- 23.8, 89.0 +/- 23.9 and 66.0 +/- 32.0, respectively (in comparison to day 1 for 4 and 8 mg/kg/day IGF 1 p < 0.05). The combined effect of 4 mg/kg/day IGF-1 and 10 IU/kg/day growth hormone (GH) was also studied in U and P animals. Two hours after the first injections of IGF-1 serum glucose levels decreased in U from 120.0 +/- 11.3 to 49.2 +/- 21.6 mg/dl, while IGF-1 plus GH decreased the glucose level from 122.0 +/- 15.5 to 81.3 +/- 24.7 mg/dl (p < 0.05 IGF-1 vs. IGF-1 + GH). The hypoglycemic effect of IGF-1 was less expressed by long-term treatment and simultaneous administration of GH overcame the glucose-lowering effect of IGF-1 (serum glucose levels on day 11 one hour after the injections: 73.7 +/- 15.3 mg/dl with IGF-1, and 111.0 +/- 7.8 mg/dl with IGF-1 + GH). Methylprednisolone (MP) did not significantly alter the former effects of IGF-1 and GH. In summary, IGF-1 leads to hypoglycemia in control and uremic rats in a dose-dependent manner. This effect becomes less expressed after prolonged administration. GH attenuates the hypoglycemic effect of IGF-1. This suggests that the combined GH and IGF-1 treatment is more effective and less dangerous in correcting uremic growth failure. PMID- 10474023 TI - Association of angiotensin-converting enzyme gene polymorphism with lipid profiles in children and adolescents with insulin-dependent diabetes mellitus. AB - We attempted to clarify the association between angiotensin-converting enzyme (ACE) gene polymorphism and the other predictive factors for macroangiopathy in children and adolescents with uncomplicated insulin-dependent diabetes mellitus (IDDM). Sixty-three patients were divided into 3 groups according to the ACE genotypes. The lipid profiles were evaluated according to ACE genotypes. The level of lipoprotein(a) (Lp(a)) in the II genotype was significantly lower than that in groups with the D allele. Lp(a) significantly correlated with apo B/apo A I (p < 0.001, r = 0.63) and atherogenic index (AI = (total cholesterol - high density lipoprotein cholesterol)/high-density lipoprotein cholesterol; p = 0. 004, r = 0.36). We suggest that the D allele may affect the level of Lp(a) and the other lipid profiles in IDDM. PMID- 10474024 TI - Expression of N-acetyl galactosaminylated and sialylated glycans by metastases arising from primary breast cancer. AB - This study examines the Helix pomatia lectin (HPA) binding characteristics of metastases arising from primary breast cancer, and compares HPA binding patterns with binding of Dolichos biflorus lectin (DBA), Limax flavus lectin (LFA), and a monoclonal antibody against the Tn epitope. Of 81 blocks of metastases in a range of tissues, taken at autopsy from 46 individuals, 79% were HPA positive. No site specificity with regard to HPA binding was observed. Both HPA-positive and negative tumour deposits were seen within a single individual. HPA-positive tumours were commonly negative for binding of sialic acid specific lectin LFA (86% were negative). Binding patterns of alpha-GalNAc specific HPA and DBA, and a monoclonal antibody against Tn epitope (GalNAc-O-Ser/Thr) were markedly different. PMID- 10474025 TI - Cyclosporin A upmodulates the alpha-subunit of the interleukin-2 receptor and the metastatic ability of murine B16F10 melanoma cells. AB - In the present study, the effect of in vitro cyclosporin A (CsA) treatment on IL 2R expression and the metastatic behavior of B16F10 melanoma cells has been reported. CsA treatment was found to increase the percentage of B16F10 cells expressing the alpha-subunit of IL-2R on the cell surface and also at the mRNA level. Moreover, CsA treated B16F10 cells also express the beta-subunit of IL-2. In vivo experiments showed that CsA increases the affinity of B16F10 metastazing cells for the liver and decreases that for the lung. CsA modulated the expression of MHC class I and class II antigens, but no significant differences in the resistance of CsA-treated B16F10 cells to NK lysis were observed. Finally, proliferation of B16F10 cells in the presence of several doses of CsA did not vary and CsA increased the amount of IL-1beta mRNA expression. These results suggest that CsA, through the modulation of cytokines and MHC antigen expression on B16F10 cells, could have an effect upon the metastatic progression of the B16F10 melanoma. PMID- 10474026 TI - Expression of matrix metalloproteinases and tissue inhibitor of matrix metalloproteinases in non-small-cell lung cancer. AB - Expression of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinases (TIMP)-1, and TIMP-2 was studied in non-small-cell lung cancer (NSCLC). Activity of MMP-2 and MMP-9 by gelatin zymography and expression of MMP 2, MMP-9, TIMP-1, and TIMP-2 mRNAs were examined in 11 lung cancer cell lines which included six small-cell lung cancer (SCLC) cell lines. Localization of MMP 2, MMP-9, TIMP-1, and TIMP-2 was examined by immunohistochemistry in 43 resected NSCLC (22 adenocarcinomas, 17 squamous cell carcinomas, 4 large cell carcinomas) using specific anti-human monoclonal antibodies. Expression of MMP-2 mRNA was detected in 5 (100%), MMP-9 in 1 (20%), TIMP-1 in 4 (80%), and TIMP-2 in 5 (100%) of 5 NSCLC cell lines examined. MMP-2 gelatinolytic activity also was detected in all five NSCLC cell lines, whereas MMP-9 activity was detected in only one cell line. In 43 patients, MMP-2, MMP-9, TIMP-1, and TIMP-2 immunoreactivity was demonstrated in 19 (44%), 9 (21%), 15 (35%), and 29 (67%) excised tumors, respectively. All stromal fibroblasts in tumor samples stained positive for MMP 2. There was a correlation between TIMP-2 immunoreactivity and disease stage (42% stage I versus 88% stages II, III, and IV) (p = 0.0024). Both cancer cell lines and NSCLC tumor samples frequently expressed MMP-2, MMP-9, TIMP-1, and TIMP-2; MMP-2 in particular was highly expressed in malignant cells and surrounding fibroblasts. These findings suggest that MMP-2 plays a more important role in invasion of NSCLC than MMP-9 and that TIMP-2 may have clinical relevance in NSCLC. PMID- 10474027 TI - Increased tumorigenicity and invasiveness of C6 rat glioma cells transfected with the human alpha-2,8 sialyltransferase cDNA. AB - Gangliosides are thought to be involved in tumor cell proliferation, migration and invasiveness as so far demonstrated by the addition of exogenous gangliosides to the culture medium. To better understand the direct influence that alterations in ganglioside synthesis can exert on these functional aspects of cell biology, in the present study, we investigated the behaviour of C6 rat glioma cells after stable transfection with the human CMP-NeuAc:NeuAcalpha2-3Galbeta1-4GlcCer alpha2,8-sialyltransferase (SAT-II, EC 2.4.99.8) gene. The enzyme synthesizes ganglioside GD(3) by adding a sialic acid residue to ganglioside GM(3). Stable transfection of the constructs into C6 cells and expression of the human SAT-II gene were evaluated using PCR and RT-PCR amplification, respectively. Qualitative and quantitative analysis of the ganglioside profile was performed by conventional HP-TLC and identity of de novo synthesized species was assessed by TLC immunostaining. Results show that whereas C6 parental cells and C6 cells transfected with the empty expression vector synthesize, almost exclusively, ganglioside GM(3), de novo synthesis of GD(3) is clearly observed in clones expressing the alpha2,8-sialyltransferase. Subcutaneous grafting in athymic nude mice of cells expressing high levels of GD(3) induces tumors growing faster and more aggressively than controls. In in vitro assays, the same cells demonstrate increased proliferation rate, motility and invasiveness. Chemotaxis and chemoinvasion were assayed using the modified Boyden chamber. Data obtained suggest that endogenously neosynthesized GD(3) is able to modify proliferation rate, motility and invasion of C6 rat glioma cells, enhancing the features of malignancy of this tumor cell line. PMID- 10474028 TI - Serum obtained from rats after partial hepatectomy enhances growth of cultured colon carcinoma cells. AB - Tumour-bearing rats were randomized to a 70% partial hepatectomy or a sham operation. At days 1, 3 or 14, portal and systemic serum was obtained and colon carcinoma cells were cultured in the presence of 5, 10, 20 or 50% serum. Proliferation and epidermal growth factor receptor (EGFr) expression was measured in tumour cells. Proliferation was 25-40% higher in tumour cells cultured with portal serum after hepatectomy than after sham operation when using serum obtained at day 3, but not days 1 and 14 after operation. In cultures with serum obtained at day 14 after operation CC 531 cells showed a 30% higher proliferation rate with systemic hepatectomy serum than CC 531 cells with sham systemic serum. These effects were not mediated by a change in EGFr mRNA and protein levels as the used colon carcinoma cells did not reveal EGFr activity by any of the three detection methods used. PMID- 10474029 TI - Tropomyosin: an invertebrate pan-allergen. AB - Among food allergens, crustaceans, such as shrimp, crab, crawfish and lobster, are a frequent cause of adverse food reactions in allergic individuals. The major allergen has been identified as the muscle protein tropomyosin. This molecule belongs to a family of highly conserved proteins with multiple isoforms found in both muscle and nonmuscle cells of all species of vertebrates and invertebrates. Its native structure consists of two parallel alpha-helical tropomyosin molecules that are wound around each other forming a coiled-coil dimer. Allergenic tropomyosins are found in invertebrates such as crustaceans (shrimp, lobster, crab, crawfish), arachnids (house dust mites), insects (cockroaches), and mollusks (e.g. squid), whereas vertebrate tropomyosins are nonallergenic. Studies of cross-reactivities among crustaceans and the high degree of sequence identity among them suggest that tropomyosin is probably the common major allergen in crustaceans. Furthermore, immunological relationships between crustaceans, cockroaches and housedust mites have been established and may suggest tropomyosin as an important cross-sensitizing pan allergen. PMID- 10474030 TI - The importance of serine proteinases as aeroallergens associated with asthma. AB - Penicillium and Aspergillus species have been identified as prevalent indoor airborne fungi that are associated with extrinsic bronchial asthma. We have recently analyzed the IgE-binding components in 8 prevalent Penicillium and Aspergillus species (P. citrinum, P. notatum, P. oxalicum, P. brevicompactum, A. fumigatus, A. flavus, A. oryzae and A. niger) by immunoblotting and N-terminal amino acid sequence analysis. Our results show that the alkaline and/or vacuolar serine proteinases are the major allergens in these prevalent fungal species. IgE cross-reactivity among these major allergens was also detected. Results obtained provide an important basis for clinical allergy. In addition, monoclonal antibodies against alkaline and/or vacuolar serine proteinase allergens have been generated. These antibodies can be applied for the standardization of allergenic extracts. PMID- 10474031 TI - Selective cloning of peanut allergens, including profilin and 2S albumins, by phage display technology. AB - BACKGROUND: Peanut kernels contain many allergens able to elicit IgE-mediated type 1 allergic reactions in sensitized individuals. Sera from sensitized patients recognize variable patterns of IgE-binding proteins. The identification of the IgE-binding proteins of peanut extract would faciliate improvement of diagnostic and immunotherapeutic approaches as well as development of sensitive test systems for the detection of hidden peanut allergens present as additives in various industrial food products and the investigation of their stability during processing of food products. METHODS: We applied the pJuFo cloning system based on the phage surface display of functional cDNA expression products to clone cDNAs encoding peanut allergens. Sera (n = 40) of peanut-allergic individuals were selected according to case history, radioallergosorbent test and immunoblot analysis to demonstrate IgE binding towards the newly identified recombinant allergens. RESULTS: In addition to the known allergens Ara h 1 and Ara h 2 we were able to identify four allergens with estimated molecular weights of 36, 16, 14.5 and 14 kDa. Three of them formally termed Ara h 4, Ara h6 and Ara h 7 show significant sequence similarities to the family of seed storage proteins and the fourth (Ara h 5) corresponds to the well-known plant allergen profilin. Immunoblotting of the six expressed recombinant allergens with 40 patients sera shows 14 individual recognition patterns and the following frequency of specific IgE binding: Ara h 1 was recognized by 65%, Ara h 2 by 85%, Ara h 4 by 53%, Ara h 5 by 13%, Ara h 6 by 38% and Ara h 7 by 43% of the selected sera. CONCLUSIONS: All of the selected peanut-positive sera can detect at least one of the six identified recombinant allergens which can be used to establish individual patients' reactivity profiles. A comparison of these profiles with the clinical data will possibly allow a further insight into the relationship between clinical severity of the symptoms and specific IgE levels towards the six peanut allergens. PMID- 10474033 TI - Alkaline serine proteinase is a major allergen of Aspergillus flavus, a prevalent airborne Aspergillus species in the Taipei area. AB - BACKGROUND: Aspergillus species are prevalent indoor airborne fungi and have been identified to be a causative agent of human allergic disorders. In the present study, we identified, purified and characterized the allergen(s) from Aspergillus flavus, a predominant airborne Aspergillus species in the Taipei area. METHODS: The IgE-binding components of A. flavus were identified by SDS-PAGE immunoblotting with sera from asthmatic patients. The N-terminal amino acid sequences of the major allergens were determined by Edman degradation. The allergenic cross-reactivity among allergens from different fungi was analyzed by immunoblot inhibition using sera from asthmatic patients. The detected major allergen was purified from the culture medium. It was further characterized in terms of its N-terminal amino acid sequence, its IgE-binding activity and its enzymatic activity. RESULTS: The results of the immunoblot analysis indicate that a 34-kD component that has high IgE-binding (63%) frequency is a major allergen of A. flavus. The N-terminus of this 34-kD major allergen (GLTTQKSAP) has high sequence identity with that of the 34-kD alkaline serine proteinase major allergen of A. oryzae. Results from immunoblot inhibition studies indicate that IgE cross-reactivity occurs among the 34-kD major allergens of A. flavus, A. fumigatus and Penicillium citrinum. The 34-kD major allergen of A. flavus was purified from the culture medium by ammonium sulfate precipitation and DEAE ion exchange chromatography. The N-terminal amino acid sequence of the purified allergen (Asp fl 13) is identical to that determined previously for the 34-kD major allergen in the crude extract of A. flavus. The IgE immunoblot reactivity to the 34-kD major allergen in the crude extract can be dose-dependently inhibited by the purified Asp fl 13. The degree of IgE binding to the 34-kD major allergen in the crude extract correlates with that of the purified Asp fl 13 in sera of 8 asthmatic patients. The purified Asp fl 13 has proteolytic activity with casein as substrate at pH 8.0. This enzymatic activity can be inhibited by either phenylmethylsulfonyl fluoride or diethylpyrocarbonate. CONCLUSION: Our results suggest that the 34-kD alkaline serine proteinase is a major allergen of A. flavus. There was IgE cross-reactivity among allergens of A. flavus, A. fumigatus and P. citrinum. PMID- 10474032 TI - Identification and characterisation of two allergens from the dust mite Acarus siro, homologous with fatty acid-binding proteins. AB - BACKGROUND: Dust mites are a major cause of allergic disease worldwide. The dust mite Acarus siro is an inducer of occupational allergy among farmers, but sensitisation has also been found in non-farming populations. METHODS: A degenerate primer was designed to the N-terminal amino acid sequence of a 15-kD IgE-binding protein in A. siro extract. The cDNA sequence was obtained by using reverse transcriptase polymerase chain reaction, standard cloning and sequencing techniques. The protein was expressed in Escherichia coli with a 6-histidine tag at its C-terminus. Immunoblotting of the recombinant protein and whole extract was performed using patient sera. RESULTS AND CONCLUSION: 15 and 17-kD allergens were identified in a fraction of A. siro extract. The cDNA of the 15-kD allergen was isolated, cloned and sequenced and the allergen was expressed as a recombinant protein. The calculated molecular weight of the cDNA-encoded protein is 14.2 kD. The predicted amino acid sequence has one potential N-glycosylation site at position 4-6 and a cytosolic fatty acid-binding protein signature at position 5-22. The protein has 64% sequence identity with Blo t 13, an allergen from the dust mite Blomia tropicalis, as well as homology with several other fatty acid-binding proteins (FABPs) from different organisms. The allergen was named Aca s 13 and was recognised strongly by 3 of 13 (23%) of the subjects investigated. The amino acid sequence of the 17-kD protein was partly determined and it also showed high sequence homology with Blo t 13 and FABPs. PMID- 10474034 TI - Anaphylactic response to parasite antigens: IgE and IgG1 independently induce death in Trichinella-infected mice. AB - The response of animals infected with different Trichinella species (T. spiralis, T. britovi, T. pseudospiralis) to antigens of different Trichinella species was evaluated in outbred mice (CD1) and inbred mice (BALB/c, C3H, SJL, C57BL/6). In mice deficient for IgE or IgG1, T. spiralis only was used. In homologous conditions (i. e., same Trichinella species for antigens and infectious agent) mortality was very high (up to 100% in SJL and C57BL/6). The anaphylactic response in heterologous conditions was much lower and death was observed only in C57BL/6 and SJL mice. Experiments with deficient mice have shown that both specific IgE and IgG1 independently induce a fatal anaphylactic response. PMID- 10474035 TI - Time course of pharmacological modulation of peak eosinophilic airway inflammation after mite challenge in guinea pigs: a therapeutic approach. AB - BACKGROUND: It is well known that eosinophilic airway inflammation develops after allergen challenge in sensitized humans and animals. However, the detailed time course of suppression of early eosinophilic airway inflammation by pharmacological agents given just after challenge has not been discussed. Therefore, we aimed to evaluate the time course relationship of the suppression of peak eosinophilia by anti-cytokines and pharmacological agents given several hours after the aerosol challenge by a therapeutic approach. METHODS: We used crude mite extract as an allergen to create a sensitization and inhalation challenge, and performed bronchoalveolar lavages (BAL) after the inhalation challenge to observe the degree of eosinophilic airway inflammation in guinea pigs. Various anti-cytokines (anti-IL-3 and anti-IL-5) and pharmacological agents (dexamethasone, theophylline, and roxithromycin) were given within several hours after the acute aeorosol challenge to evaluate the suppressive effect on peak eosinophilia in BAL fluid, which occurred 24 h after the challenge. RESULTS: Our results show that anti-IL-5 and dexamethasone, given within 4 and 8 h after the inhalation challenge, respectively, inhibit the acute allergen-induced peak eosinophilia in BAL fluid. However, anti-IL-3, theophylline, and roxithromycin had no effect on peak eosinophilic airway inflammation after challenge. CONCLUSION: These observations suggest that several hours are needed to complete the process of cytokine-induced recruitment of eosinophils from the blood to the airways after acute allergen challenge. This may be the optimal time to administer anti-cytokines and dexamethasone to attenuate the subsequent eosinophilic airway inflammation after acute allergen-induced asthmatic attacks. PMID- 10474036 TI - Pathophysiological features of the nasal mucosa in patients with idiopathic rhinitis compared to allergic rhinitis. AB - BACKGROUND: The literature on abnormality of vasomotor responses of the nasal mucosa to cold stimulation of the skin in idiopathic rhinitis is conflicting. The objective of this study was to elucidate pathophysiological features of the nasal mucosa in idiopathic rhinitis compared to allergic rhinitis. METHODS: The following were studied in patients with idiopathic rhinitis and allergic rhinitis and in normal controls: (1) threshold of the nasal reaction to histamine; (2) inflammatory cells in nasal lavage and scraped nasal mucosal epithelium, and (3) nasal vasomotor response to cold stimulation of the feet evaluated by acoustic rhinometry. RESULTS: Inflammatory cells were not found to be involved in idiopathic rhinitis. Nasal reactivity to histamine was significantly enhanced in patients with idiopathic rhinitis compared to normal controls, but was significantly lower compared to those with allergic rhinitis. The most prominent finding in idiopathic rhinitis was nasal mucosal swelling induced by cold stimulation of the feet. While in normal controls, cold stimulation of the feet caused mucosal contraction due to sympathetic excitation, sympathetic nasal vasomotor response in idiopathic rhinitis patients was significantly inhibited and caused mucosal swelling and enhanced nasal secretion. Mucosal reactions observed in allergic rhinitis were between those observed in idiopathic rhinitis and in normal controls. Cold stimulation of the feet increased systolic blood pressure by 5-15 mm Hg, but the degree of increase observed in the 3 groups was almost equal. CONCLUSIONS: The above findings indicate that patients with idiopathic rhinitis have abnormalities that inhibit sympathetic reactions and enhance parasympathetic vasomotor response at peripheral levels, possibly in the nasal mucosa. PMID- 10474037 TI - Distinct expression of RANTES and GM-CSF by lipopolysaccharide in human nasal fibroblasts but not in other airway fibroblasts. AB - Eosinophil infiltration of tissue is a hallmark of nasal polyposis and asthma in both atopic and nonatopic patients. Structural cells like airway fibroblasts are a rich source of cytokines and inflammatory mediators. In order to verify whether airway fibroblasts play a role in eosinophilic infiltration, we investigated the release of eosinophil chemotactic and activating factors from airway fibroblasts when stimulated with nonallergenic exogenous agents such as endotoxin (lipopolysaccharide; LPS). Using a number of primary human airway tissue-derived fibroblast lines, we demonstrated that LPS could induce the gene expression and production of RANTES (regulated and normal T cell expressed and presumably secreted) and granulocyte/macrophage colony-stimulating factor (GM-CSF) only in nasal but not in pharyngeal, tracheal, bronchial, and lung fibroblasts. This selective responsiveness of nasal fibroblasts to LPS was time and dose dependent. These findings suggest that nasal fibroblasts may play an important role in the recruitment and activation of eosinophils into nasal polyps through the release of RANTES and GM-CSF. PMID- 10474038 TI - Natural rubber latex allergy is not a cause of sudden infant death. AB - BACKGROUND: The causes for sudden infant death (SID) remain unclear. As infants can become sensitized to NRL allergens by pacifiers and latex mattresses, we tried to establish whether there is a relationship between SID and natural rubber latex (NRL) allergy. METHODS: We determined NRL-specific IgE concentrations in 112 unselected cases of SID by the CAP-FEIA method. RESULTS: NRL-specific IgE could be detected only in 1 sample (0.64 kU/l; CAP class 1). CONCLUSIONS: We conclude that NRL allergy is not a cause of SID. PMID- 10474039 TI - MDR1 gene expression in normal and atherosclerotic human arteries(1). AB - Recent studies have shown that a membrane p-glycoprotein, encoded by MDR1 gene, is involved in the transport of free cholesterol from the plasma membrane to endoplasmic reticulum, the site of cholesterol esterification by acyl CoA:cholesterol acyltransferase (ACAT). Moreover, results deriving from our previous studies have shown that the rate of cell proliferation was positively correlated with cholesteryl ester levels as well as with ACAT and MDR1 gene expression. In this study, lipid content and the expression of the genes involved in cholesterol metabolism such as hydroxy-methylglutaryl coenzyme A reductase (HMGCoA-R), low-density lipoprotein receptor (LDL-R), ACAT and MDR1 have been investigated in control and atherosclerotic arteries. The results have shown that the levels of cholesteryl ester increase with the age of cadaveric donors in arteries prone to atherosclerosis (abdominal aorta, superficial femoral artery) and become predominant in advanced atherosclerotic lesions. The mRNA levels of ACAT and MDR1 showed the same age correlation, reaching the highest values in atherosclerotic specimens. These results suggest that MDR1 may be involved in the accumulation of intracellular cholesterol ester levels found in atherosclerotic lesions. Moreover, the levels of HMGCoA-R, LDL-R and ACAT gene expressions progressively increased with the age of cadaveric donors; conversely, in atherosclerotic specimens, the mRNA levels of HMGCoA-R and LDL-R drastically decreased while ACAT gene expression reached its maximum. These findings suggest a reactivation of normal homeostatic regulation of cholesterol in advanced and complicated lesions. PMID- 10474040 TI - Expression of tie1 and tie2 proteins during reendothelialization in balloon injured rat carotid artery. AB - The novel endothelial cell tyrosine kinase receptors, Tie1 and Tie2, are essential for vascular development and remodeling in the embryo but little is known regarding the regulation of their expression and their role in the maintenance and repair of the adult vascular system. We examined the expression of Tie1 and Tie2 in normal vessels and during reendothelialization following balloon injury of the adult rat carotid artery. Tie proteins were detected in quiescent endothelial cells of the adult rat carotid artery. Tie1 and Tie2 proteins were also detected in human and rat platelets. A weak expression of Tie1 and Tie2 proteins was detected in young endothelial cells which sparsely repopulated the denuded surface by day 14. Protein levels increased in the confluent layer of endothelial cells by day 28. Based on these observations, we tested whether Tie1 and Tie2 mRNA and protein levels are regulated by cell density. Tie1 and Tie2 expression significantly increased with higher density in cultured human endothelial cells, and this upregulation required cell-cell interaction. These data suggest that Tie1 and Tie2 may play a role in the maintenance and repair of the adult vascular system and that the expression of these proteins is regulated by cell density. PMID- 10474041 TI - Leukocyte margination in alveolar capillaries: interrelationship with functional capillary geometry and microhemodynamics. AB - The pulmonary capillary microvasculature harbors a large pool of intravascularly marginated leukocytes. In this study, we investigated the interrelationship of leukocyte margination with characteristics of functional capillary geometry and microhemodynamics in alveolar capillary networks. In 22 anesthetized rabbits we assessed functional capillary density, average capillary length, red blood cell velocity and leukocyte kinetics in alveolar capillary networks in vivo by intravital fluorescence microscopy. In alveolar wall areas of 12,800 +/- 1,800 microm(2), we detected 3.6 +/- 0.5 sticking leukocytes and 21.0 +/- 1.9 functional capillary segments with an average capillary length of 35.7 +/- 2.1 microm. We calculated that approximately 15% of functional capillary segments are blocked by marginated leukocytes. Leukocyte margination was predominantly observed in capillary networks characterized by a high functional capillary density, short capillary segments and low red blood cell velocities. The multitude of interconnected capillary channels in these networks may allow alveolar blood flow to bypass marginated leukocytes. Hence, this interrelationship may be relevant for maintenance of adequate alveolar perfusion and low capillary network resistance despite excessive leukocyte margination in the pulmonary microvasculature. Local microhemodynamic factors may play a regulatory role in the spatial distribution of leukocyte margination. PMID- 10474042 TI - Leukocyte sequestration in pulmonary microvessels and lung injury following systemic complement activation in rabbits. AB - Inflammatory reactions are associated with sequestration of leukocytes in the lung. Complement activation leads to accumulation of leukocytes in alveolar septa and alveoli, to lung edema and hemorrhage. Although in organs other than the lung leukocytes interact with the vascular endothelium only in postcapillary venules, alveolar capillaries are considered to be the site of leukocyte sequestration in the lung. However, pulmonary venules and arterioles have not been investigated systematically after complement activation so far. A closed thoracic window was implanted in anesthetized rabbits; leukocytes and red blood cells were stained, and the movement of these cells was measured in superficial pulmonary arterioles, venules and alveolar capillaries using fluorescence video microscopy before and 30 and 60 min after infusion of cobra venom factor (CVF). Erythrocyte velocity and macrohemodynamic conditions did not change after CVF infusion and were not different from the sham-treated controls. The number of sticking leukocytes increased significantly compared to baseline and control: by 150% in arterioles and in venules and by 740% in alveolar capillaries within 60 min after CVF infusion. The width of alveolar septa in vivo was significantly enlarged after CVF infusion, indicating interstitial pulmonary edema. At the end of the experiments, myeloperoxidase activity was higher in the CVF group, showing leukocyte sequestration in the whole organ. It is concluded that complement activation by CVF induces leukocyte sequestration in lung arterioles, venules and alveolar capillaries and leads to mild lung injury. PMID- 10474043 TI - Characterization of major phosphoproteins in the cGMP-mediated protein phosphorylation system of vascular smooth muscle membranes. AB - G(0) (215-250 kD) and G(1) (120-140 kD), the unidentified major phosphoproteins in the cGMP-mediated protein phosphorylation system of vascular smooth muscle membranes, were compared for biochemical and immunological properties with the type 1 inositol 1,4, 5-trisphosphate receptor (InsP(3)R, 240 kD) and the myosin binding subunit (MBS, 138 kD) of myosin phosphatase, both of them substrates for cGMP-dependent protein kinase. Two microsomal proteins that were immunoreactive with antibodies to InsP(3)R and MBS were detected, and comigrated with G(0) and G(1), respectively, on SDS-PAGE. When thiophosphorylated G(0) and G(1) were subjected to immunoprecipitation, MBS antibody induced the precipitation of a 138 kD phosphoprotein, but did not significantly affect the amount of G(1) remaining in the supernatant, while InsP(3)R antibody precipitated G(0) almost completely. Unexpectedly, InsP(3)R antibody coprecipitated a large portion of G(1), which did not cross-react with either antibody to MBS or InsP(3)R. Just like InsP(3)R, G(0) bound to the calmodulin column in a Ca(2+)-dependent manner, and, again, a large portion of G(1) was copurified with G(0). These results suggest that G(0) is identical to InsP(3)R, while G(1) consists of several phosphoproteins, including the 138-kD protein associated with InsP(3)R as a major component. MBS is not G(1) or may represent only a minor component of it. PMID- 10474044 TI - Relaxation in different-sized rat blood vessels mediated by endothelium-derived hyperpolarizing factor: importance of processes mediating precontractions. AB - To clarify the mechanisms involved in relaxations mediated by endothelium-derived hyperpolarizing factor (EDHF), acetylcholine (ACh)-induced endothelium-dependent relaxations and hyperpolarizations were examined in the rat aorta, the main branch of the mesenteric artery (MBMA) and the first branch of the mesenteric aftery (FBMA). In the presence of 100 microM N(G)-nitro-L-arginine (L-NNA) and 10 microM indomethacin, ACh (1 nM to 100 microM) produced no relaxation in the phenylephrine-precontracted aorta. The L-NNA-resistant relaxations by ACh in MBMA precontracted with phenylephrine were eliminated in the presence of 1 microM nifedipine where contractions were independent of L-type Ca(2+) channel activation. In FBMA precontracted with phenylephrine, the L-NNA-resistant relaxations were only partially inhibited by nifedipine. When vessels had been contracted with 300 nM phorbol-12,13-dibutyrate in the presence of nifedipine, ACh-induced L-NNA-resistant relaxations were observed in FBMA only. Pinacidil produced relaxations in all different-sized blood vessels, although sensitivity was inversely related to vessel size. The extent of the ACh hyperpolarizing responses was much smaller than that by pinacidil in the aorta. The membrane potential changes by ACh and pinacidil were almost the same in FBMA. These results indicate that the contribution of EDHF to endothelium-dependent relaxations increases as the vessel size decreases. This may be partly explained by precontractile processes dependent on Ca(2+) entry through L-type Ca(2+) channels, because Ca(2+) channel deactivation seems to be involved as a major mechanism of EDHF-mediated vasorelaxations. However, EDHF may also generate vasorelaxations by an additional mechanism, probably a reduced Ca(2+) sensitivity of contractile elements, as proposed for ATP-sensitive K(+) channel openers. PMID- 10474045 TI - Inflammation in chronic venous insufficiency. Is the problem insurmountable? AB - One of the hallmarks of venous insufficiency is an elevated venous pressure. While a number of mechanisms have been proposed for vascular and parenchymal cell damage following venous pressure elevation, such as white cell infiltration, a key question remains as to what degree venous occlusion and flow interruption per se may constitute a risk factor in venous disease. To gain an insight into this mechanism, we examined the effect of venous occlusion followed by reperfusion. A draining venule (circa 50 micrometer) in the rat mesentery was occluded with a micropipette (1 h) followed by reperfusion (1 h). The procedure serves to raise the microvascular pressure to about 31 mm Hg during the occlusion while the flow is completely stopped in the local venous and capillary network. Parenchymal cell death in the mesentery was monitored by propidium iodide (PI) labeling. The number of PI-positive cells significantly increased predominantly during reperfusion. A 1-week treatment with a micronized purified flavonoid fraction (100 mg/kg/day) served to significantly reduce parenchymal cell death as well as leukocyte rolling, adhesion to postcapillary venule, and migration into the tissue both during occlusion and reperfusion. The results indicate, that even in an initially symptomless tissue, flow reduction combined with microvascular pressure elevation during venous occlusion results in tissue damage not only during reperfusion (as in arterial occlusion) but also during occlusion. PMID- 10474046 TI - Leukocyte adhesion after oxidant challenge in the hamster cheek pouch microcirculation. AB - Oral administration of S-5682 (Daflon 500 mg, 90% diosmin, 10% hesperidin) inhibits oxidant-induced increase in macromolecular permeability in the postcapillary venules of the hamster cheek pouch microcirculation. In this study, the effect of S-5682 on leukocyte-endothelium interaction was evaluated using the same experimental model. Hamsters kept on a standard diet were divided into 5 groups (n = 6) and treated orally, twice a day, with placebo (10% lactose solution), S-5682, 5, 20 or 80 mg/kg/day (suspended in 10% lactose solution) or alpha-tocopherol, 1 mg/kg/day, for 10 days prior to the oxidant challenge with tert-butylhydroperoxide (TBOOH). Topical application of TBOOH (10(-4) M for 5 min) to hamsters given acridine orange prior to TBOOH resulted in increases in the number of rolling and sticking (no movement for at least 30 s) leukocytes in postcapillary venules. No changes in the number of rolling leukocytes could be observed in the treated groups compared with the placebo group (p > 0.05). On the contrary, leukocyte adhesion was inhibited in groups treated with S-5682 (5, 20 and 80 mg/kg/day) or alpha-tocopherol: placebo 105 +/- 3/6 mm(2) (mean +/- SEM); S-5682, 5 mg/kg/day 68 +/- 3/6 mm(2) (p < 0.01), 20 mg/kg/day 55 +/- 3/6 mm(2) (p < 0.001) and 80 mg/ kg/day 39 +/- 2/6 mm(2) (p < 0.001) and alpha-tocopherol 36 +/- 1/6 mm(2) (p < 0.001). The inhibition of oxidant-induced leukocyte adhesion by S-5682 was similar to that seen for ischemia-reperfusion and the higher dose of S-5682 was as effective as alpha-tocopherol in inhibiting it. PMID- 10474047 TI - Adhesion molecule expression in postischemic microvascular dysfunction: activity of a micronized purified flavonoid fraction. AB - Ischemia and reperfusion (I/R) induces neutrophil infiltration in skeletal muscle that is localized to the ischemic region. To transmigrate at ischemic regions, granulocytes must first arrest in the postcapillary venular segment of the microcirculation. Initially, leukocytes roll along the endothelium of these venules, a weak adhesive interaction that is mediated by the selectins (L-, E-, and P-selectin). Leukocyte rolling functions to slow the neutrophil during its transit through the microcirculation, thereby allowing it to monitor its local environment for the presence of activating factors arising from the ischemic tissues. When activated, the rolling granulocyte is rendered capable of forming the stronger adhesive interactions that allow the cell to become arrested in postcapillary venules in the ischemic region. These adhesive interactions are mediated by a leukocyte glycoprotein complex designated CD11/CD18 and intercellular adhesion molecule-1 (ICAM-1) expressed on endothelial cells. The stationary neutrophil uses the gradient in concentration of soluble chemoattractants liberated from ischemic tissues as a directional cue to move from the vascular to extravascular compartment, being guided in its transit across the endothelium by interactions with platelet endothelial cell adhesion molecule-1 (PECAM-1), an adhesive molecule localized to the interendothelial cleft. This paper reviews current understanding of the mechanisms underlying the establishment of leukocyte/endothelial cell interactions in postischemic skeletal muscle in terms of specific adhesion molecules that participate in neutrophil sequestration after I/R. Discovery of the molecular determinants of neutrophil/endothelial cell adhesion has uncovered potential mechanisms whereby agents exhibiting anti-adhesive properties may act. The micronized purified flavonoid fraction (450 mg diosmin, 50 mg hesperidin) prevents I/R-induced leukocyte adhesion in skeletal muscle. This anti-adhesive effect appears to be mediated at least in part by inhibition of induced expression of ICAM-1. PMID- 10474048 TI - Neutrophil activation and mediators of inflammation in chronic venous insufficiency. AB - The effect of venous hypertension on the state of activation of leucocytes has been investigated in patients with venous disease and control subjects. Leucocytes become 'trapped' in the circulation of the leg during periods of venous hypertension produced by sitting or standing. This is greater in the limbs of patients with chronic venous disease than controls. Studies of the plasma levels of neutrophil granule enzymes show that these are increased during periods of venous hypertension, suggesting that this causes activation of the neutrophils. Investigation of the leucocyte surface ligand CD11b shows that the more activated neutrophils and monocytes are sequestered during venous hypertension. Measurement of plasma levels of the soluble parts of the vascular (VCAM), intercellular (ICAM) and endothelial leucocyte (ELAM) adhesion molecules show that these are all elevated in patients with chronic venous disease compared to controls. Following 30 min of venous hypertension produced by standing, these levels are further increased. These data suggest that venous hypertension causes neutrophil and monocyte activation, which in turn causes injury to the endothelium. I believe that this may be the mechanism that initiates the pathological processes which lead to venous ulceration. It has recently been shown that the venotonic drug Daflon 500 mg (450 mg diosmin, 50 mg hesperidin, Servier, France) influences these processes. Surface expression of CD62L is reduced in neutrophils and monocytes, and plasma levels of soluble endothelial adhesion molecules are reduced. These observations may explain the anti inflammatory effects of Daflon 500 mg. PMID- 10474049 TI - Clinical efficacy of micronized purified flavonoid fraction: an overview. AB - Flavonoids are widely used for the treatment of chronic venous insufficiency (CVI). In clinical trials, micronized purified flavonoid fraction (MPFF, 450 mg diosmin plus 50 mg hesperidin, Daflon 500 mg) has demonstrated its activity in CVI by improving venous tone and vein elasticity assessed by plethysmography. Randomized, double-blind, placebo-controlled clinical studies have shown an improvement in signs and symptoms related to CVI and a decrease in leg circumferences at the levels of the ankle and calf. The effect of MPFF on microcirculatory parameters suspected of participating in the pathophysiological process of venous ulceration has been investigated in patients. These include hemorheological parameters and transcutaneous oxygen tension measurements, which were shown to improve after treatment with MPFF. Finally, a randomized double blind, placebo-controlled clinical study has shown that MPFF treatment for 2 months, in addition to standard compression therapy, accelerates leg ulcer healing in patients with ulcers elderly adults) and task (single-task > dual-task) effects were observed for performance accuracy and speed measures as well as for N400/P600 ERP waveform components elicited by 'old/new' word recognition and P300 ERP indices elicited by auditory target detection. The effects of smoking history were limited to the P600 component, which showed faster latencies in elderly smokers than elderly nonsmokers and young nonsmokers. Young smokers exhibited longer latencies than young nonsmokers. The results were discussed in relation to normal and pathological cognition in the elderly. PMID- 10474065 TI - The acute effects of the noradrenaline reuptake inhibitor Org 4428 on EEG sleep in healthy volunteers. AB - Drug-induced improvement of depression may be mediated by changes in sleep physiology. In earlier studies on sleep EEG changes during treatment with antidepressants in depressed patients it could not be excluded that sleep disruptions and changes in the amount and distribution of REM sleep play a role in the changes in the sleep EEG. Therefore knowledge of the effects of antidepressants on the sleep EEG in healthy subjects with non-disturbed baseline sleep is necessary. In a three-way cross-over study in 12 healthy volunteers two single doses of Org 4428 (a highly specific noradrenaline reuptake inhibitor), 25 and 100 mg, were compared with placebo. Sleep EEGs were visually analysed and EEG power of non-REM sleep was measured. The results indicate that sole noradrenaline reuptake inhibiting activity is a potent mechanism to affect sleep polygraphic variables in an antidepressant-like way, i.e. REM sleep suppression and lengthening of REM latency. Despite the increase in the duration of non-REM sleep, i.e. stage 2, no significant changes in EEG power in the range 1-15 Hz were found. Therefore, the acute REM sleep suppression of Org 4428 did not result in a simultaneous reduction of EEG power during non-REM sleep. To date, these and earlier results indicate that most drugs with antidepressant properties affect REM sleep variables consistently, whereas their effect on both sleep polygraphic and EEG power variables in non-REM sleep is unpredictable. PMID- 10474066 TI - The effect of nitric oxide on the contractile tone of Muller cells. AB - The effect of nitric oxide (NO) on the contractile tone of Muller cells was investigated. Muller cells, isolated from the retina of chick embryos, were cultured on thin sheets of silicone. S-Nitro-N-acetyl-DL-penicillamine (SNAP), an NO donor, was applied in the presence or the absence of carboxyl phenyltetramethylimidazole oxide (C-PTIO), an NO scavenger. The contractile tone of Muller cells was assessed by the extent of wrinkles created on the silicone sheets. The change of contractile tone was evaluated quantitatively by digitizing the photograph before and after the application of SNAP. Relaxation of wrinkles was induced by SNAP. C-PTIO inhibited the SNAP-induced relaxation of wrinkles. These results suggest that NO affects the contractility of embryonic Muller cells and could thus modulate ocular development. PMID- 10474067 TI - Effects of intravitreal injection of botulinum toxin on the electroretinogram of rats. AB - The retinal toxicity of botulinum toxin A (BTA) was electroretinographically studied in rats. Sixteen rats were injected intravitreally with 10 ng of BTA. A response-amplitude series was recorded before and 1, 6, 13 and 21 days after the injection of BTA. BTA did not alter the amplitude and the peak latency of the a wave. The amplitude of the b-wave was not changed except for 2 rats, in which the b-wave was diminished. The peak latency of the b-wave was significantly prolonged after injection of 10 ng BTA (p < 0.05). Except for these latter 2 rats, the results indicated that the dosage used therapeutically appears to have no deleterious effect on retinal integrity or function at least in the short term, but multiple injections or higher doses of BTA could alter retinal function. PMID- 10474068 TI - The human fetal retinal pigment epithelium: A target tissue for thyroid hormones. AB - Thyroid hormone (T(3)) has previously been shown to regulate visual function in experimental animals and humans. To determine if T(3) exerts direct effects on retinal function, cultured human fetal retinal pigment epithelial (RPE) cells were tested for the presence of thyroid hormone receptors (TRs) and T(3) responses. Using TR-isoform-specific reverse-transcriptase polymerase chain reaction techniques, mRNA was detected for alpha1, alpha2 and beta1 TR isoforms. Immunohistochemistry using a polyclonal antibody that simultaneously recognizes alpha1, alpha2 and beta1 TRs showed nuclear staining of the fetal RPE. Specific binding of (125)I-T(3) to RPE cell nuclear extracts was detected, and Scatchard analysis revealed a K(d) of 110 pM. To determine if RPE cells can respond to T(3), hyaluronic acid (HA) levels in cell culture media were measured after 2, 4 or 6 days of growth in medium containing 10(-7) M T(3). T(3) inhibited accumulation of HA in the cell culture medium of RPE cells. This effect was not evident at 2 days, but at 4 days there was 42.8% less HA in cell culture medium of RPE cells grown in 10(-7) M T(3) (p < 0.01, t test). The effect persisted through 6 days, when there was 46.3% less HA in cell culture medium of RPE cells grown in 10(-7) M T(3) (p < 0.001, t test). The data indicate that human fetal RPE cells are a direct target for thyroid hormones. PMID- 10474069 TI - In vitro testing of antioxidants and biochemical end-points in bovine retinal tissue. AB - Lipid peroxidation in aliquots of bovine retina (without rod outer segments, ROS), purified ROS and retinal pigment epithelium (RPE) was initiated with 5 mM ferric iron and 80 mM ADP. After 30 min of oxidation at 37 degrees C, the concentration of thiobarbituric-acid-reacting substances (TBARS) which approximates lipid hydroperoxide (LHP), increased in the ROS from 2.0 +/- 3.6 to 90.2 +/- 34.5 nmol malondialdehyde (MDA)/mg protein and in the RPE from 0.54 +/- 0.2 to 51.5 +/- 15.8 nmol MDA/mg protein. Sixteen lipid and aqueous antioxidants (AOX) from natural or synthetic sources, including five flavonoids, were evaluated for their ability to inhibit the oxidative reaction. Palm-oil-derived vitamin E showed significant protection in retina, ROS and RPE (64, 68 and 74%), respectively. Of the flavonoids tested, good protection in the retina was found at 10(-5) M for epigallocatechin gallate (50%) and at 50 ng/ml for pycnogenol (61%) and catechin (52%). When catechin and palm oil vitamin E, catechin and coenzyme Q(10) or coenzyme Q(10) and pycnogenol were combined, the individual effect was enhanced. TBARS as an indirect measure of LHP level and hemoglobin methylene blue determination for direct LHP were used as alternative end-point determinations of lipid peroxidation. These measure different aspects of AOX reactions. The results demonstrate the usefulness of an in vitro model system that can rapidly and accurately determine the capacity of a single AOX against lipid peroxidation or be used to show synergistic efficacy. PMID- 10474070 TI - Fetal calf serum protects cultured porcine corneal endothelial cells from endotoxin-mediated cell damage. AB - In corneal organ culture, a contamination of sterile culture media with endotoxin is frequently found. Thus, we investigated if the presence of endotoxin affects the viability of cultured porcine corneal endothelial cells. Endotoxin in high concentrations caused morphological cell changes in porcine corneal endothelial monolayer cultures, delayed proliferation and decreased cellular esterase activity of porcine corneal endothelial cells in vitro. The toxic effect of endotoxin was modulated by the fetal calf serum content of the medium, the concentration of endotoxin and the incubation time. PMID- 10474071 TI - Topically applied water extract of propolis to suppress corneal neovascularization in rabbits. AB - PURPOSE: Propolis, a natural honey bee hive product, has anti-inflammatory and antioxidative properties. We aimed to assess the possible contribution of topically applied propolis to the suppression of corneal neovascularization (CNV). METHODS: The effect of a water extract of propolis (WEP) 1% drops (group 1) in comparison with dexamethasone 0.1% (group 2) and saline (group 3) on CNV was tested in rabbit corneas injured by silver nitrate cauterization. The extent of CNV was quantitated as the area of CNV and the percent area of CNV for each cornea of the three groups (12 right eyes per group) in the first week of the treatment. The mean percent CNV was used for statistical analysis. RESULTS: The corneas treated with the topical WEP 1% had an almost equal percent CNV as compared with the corneas treated with topical dexamethasone 0.1% and had less percent CNV than the control eyes. The quantitative analysis in groups 1, 2 and 3 revealed that the mean percent CNV was 41.0 +/- 14.1, 39.4 +/- 11.0 and 56.9 +/- 18.4, respectively. The differences between both groups 1 and 3 as well as groups 2 and 3 were statistically significant (p = 0.02 and p = 0.01, respectively), whereas the difference between groups 1 and 2 was not significant (p = 0.86). CONCLUSIONS: The topical application of a WEP 1% has an inhibitory effect on CNV in the rabbit's cornea. The inhibitory effect of propolis was shown to be comparable to that of topical dexamethasone 0.1%, a potent inhibitor of angiogenesis. We suggest that the effect of propolis may partially be due to its inhibitory effect on the activity of both cyclo-oxygenase and lipo-oxygenase. PMID- 10474072 TI - Effect of Na-hyaluronan on stromal and endothelial healing in experimental corneal alkali wounds. AB - The healing of stroma and endothelium after a standardized corneal alkali wound was evaluated in sodium-hyaluronan (Na-HA)-treated eyes. Alkali wounds were produced in one eye of each rabbit by applying a 5.5-mm round filter paper soaked in 1 N NaOH onto the central cornea for 60 s. Eyes were then treated with either 1% Na-HA (the treatment group) or phosphate-buffered saline (the control group) 4 times per day for 3 weeks. Endothelial wound morphometry was performed after alizarin red and trypan blue staining, while stromal healing was assessed by counting polymorphonuclear leukocytes (PMNs) and keratocytes in the central and marginal wound areas. During the early healing period the stroma treated with Na HA had less PMNs than that of the control group. The size of the endothelial defect area measured 5 days after injury was significantly smaller in the Na-HA group than in the control corneas. The present findings indicate that topically applied 1% Na-HA affects stromal and endothelial healing during the early repair process after corneal alkali wounds. PMID- 10474073 TI - Alpha-tocopherol derivatives in an experimental model of filtering surgery. AB - PURPOSE: To evaluate the efficiency and reliability of alpha-tocopherol in an experimental model of glaucoma filtering surgery. METHODS: Thirty pigmented rabbits were randomly divided into three study groups. Twenty-four hours before surgery, the animals were injected subconjunctivally with 0.5 ml of solution that depended on the group of treatment: group I (n = 10), 0.75% ethanol in balanced saline solution (BSS); group II (n = 10), 100 mg alpha-tocopherol acetate (ATA) in 0.75% ethanol in BSS; group III (n = 10), 100 mg alpha-tocopherol acid succinate (ATS) in 0.75% ethanol in BSS. The animals were followed during 30 days (intraocular pressure, IOP; filtering surgery; inflammatory reaction). RESULTS: IOPs were significantly lower in the treatment groups (ATA and ATS) than in the control group from days 7 and 10, respectively, till the end of the study. On day 7, the mean IOP in the ATA group was 15.8 versus 22.1 mm Hg in the control group. On day 10, the mean IOP in the ATS group was 15 versus 22.78 mm Hg in the control group. Filtering blebs showed statistically significant differences between the control and treated groups from day 5 to day 16. CONCLUSIONS: alpha-Tocopherol (ATA and ATS) showed better IOP control and bleb survival in this experimental model of filtering surgery. PMID- 10474074 TI - Ofloxacin levels after intravitreal injection. Effects of trauma and inflammation. AB - PURPOSE: This study was carried out to get an insight into the ofloxacin elimination after intravitreal injection in rabbits. We also studied the effects of trauma and inflammation on the vitreous ofloxacin levels after intravitreal injection of ofloxacin. METHODS: A penetrating eye injury in the right eye was inflicted on 24 rabbits and another 12 animals were used as control. A standardized intraocular inflammation was induced by intravitreal injection of a suspension of Staphylococcus aureus in half of the traumatized eyes. Ofloxacin (200 microg/0.1 ml) was injected into the midvitreous cavity of both traumatized and control right eyes, and samples were obtained at 2, 8, 24 and 48 h after injection. Drug concentrations were measured using high-pressure liquid chromatography analysis. RESULTS: Vitreous levels of ofloxacin were above the MIC(90) at 2 and 8 h in all groups for most of the common microorganisms causing endophthalmitis and also at 24 h in traumatized-infected eyes. At the second hour, the mean vitreous concentrations of ofloxacin both in traumatized and traumatized-infected eyes were lower than that in the control eyes (p < 0.05). At 8 h, the mean vitreous concentrations of ofloxacin in the traumatized and in the traumatized-infected eyes were higher than that in the control eyes (p < 0.05). At 24 h, the mean ofloxacin concentration was higher in the traumatized-infected eyes than that in control (p < 0.01) and traumatized eyes (p < 0.05), and also higher in the traumatized eyes than that in the control eyes (p < 0.05). The mean ofloxacin concentrations in the traumatized and traumatized-infected eyes were significantly higher (p < 0.01) than those in the controls at 48 h. The elimination half-life of ofloxacin in the control eyes was 5.65 h and trauma and inflammation prolonged the half-life to 9.47 and 9. 72 h, respectively. CONCLUSION: Clearance of ofloxacin is fast and appears to be reduced by trauma and inflammation. Therapeutic drug levels in traumatized-infected eyes were maintained up to 24 h. This may be an important pharmacokinetic advantage in treating endophthalmitis unless the dose used has local toxicity and allows a longer dose interval when the dose is repeated. PMID- 10474075 TI - Alpha-crystallin/lens lipid interactions using resonance energy transfer. AB - Resonance energy transfer was used to study the interaction of alpha-crystallin with lens cortex lipid vesicles. The binding of alpha-crystallin to cortex lipid vesicles and the preincubation temperature dependence of the binding were confirmed. In this study, the tryptophan of alpha-crystallin was used as the energy donor, and the fluorescence probe N-(5-dimethylaminonaphthalene-1 sulfonyl)-1, 2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt (dansyl DHPE) was chosen as the energy acceptor. Lens cortex lipid vesicles were preincorporated with dansyl DHPE. Energy transfer from the tryptophan of alpha-crystallin to dansyl DHPE was found and the energy transfer efficiency was calculated. There was a higher energy transfer efficiency between alpha crystallin and dansyl DHPE when alpha-crystallin was preincubated at 65 degrees C compared to 22 degrees C. Data confirmed the binding of alpha-crystallin to lens cortex lipid and showed that alpha-crystallin bound more closely to the surface of cortex vesicles when it was preincubated at a higher temperature. This is probably due to the exposure of hydrophobic surfaces when alpha-crystallin is preincubated at a higher temperature. PMID- 10474076 TI - The effects of high-dose corticosteroid therapy on optic nerve head blood flow in experimental traumatic optic neuropathy. AB - PURPOSE: The objective of this study was to ascertain the effect of high-dose steroid therapy in traumatic optic neuropathy using the Heidelberg retina flowmeter (HRF) to detect changes in optic nerve head blood flow. METHODS: We experimentally damaged the optic nerves of 10 white rabbits with a Hartman mosquito clamp. The first group (n = 5) was treated with intravenous dexamethasone (0.25 mg/kg) every 6 h for a 48-hour period. The second group (n = 5) served as controls. Both groups were tested prior to operation, 1 week, 1 month and 2 months after surgery. RESULTS: Experimental optic nerve damage caused a significant decrease in optic nerve head blood flow of 50.51% (p = 0.001), with a volume decrease of 46.02% (p = 0.001) and a velocity reduction of 43.12% (p = 0.002) compared to the baseline. After 2 months, in the group treated with high dose corticosteroid therapy, optic nerve head blood flow was increased by 76.90% (p = 0.012), volume by 77.53% (p = 0.012) and velocity by 47. 21% (p = 0.012) compared to the control group. CONCLUSION: The high-dose corticosteroid therapy improved the optic nerve head blood flow as demonstrated in experimental traumatic optic neuropathy. The HRF may be used to assess the therapeutic responses in traumatic optic neuropathy under variable conditions. PMID- 10474077 TI - In vivo receptor occupancy and plasma concentration of pranidipine, a potent and long-acting dihydropyridine calcium antagonist. AB - The occupancy of 1,4-dihydropyridine (DHP) receptors by pranidipine was characterized in tissues of spontaneously hypertensive rats (SHR). Oral administration of pranidipine (1 and 3 mg/kg) in SHR produced significant (26 67%) decreases in the number of specific (+)-[(3)H]PN 200-110 binding sites (B(max)) with 2- to 4-fold increases in the apparent dissociation constant (K(d)) in myocardial tissues at 1, 3 and 6 h later. In these rats, there was a reduction (16-37%) of B(max) in cerebral cortical (+)-[(3)H]PN 200-110 binding. Occupancy of myocardial DHP receptors after oral administration of pranidipine correlated well with its plasma concentration. Oral administration of nifedipine (10 mg/kg) in SHR caused significant increase in K(d) values for (+)-[(3)H]PN 200-110 binding in myocardium and cerebral cortex at 1 h later. In vivo specific (+) [(3)H]PN 200-110 binding in particulate fractions of SHR aorta was markedly (59 78%) reduced at 1, 3 and 12 h after oral administration of pranidipine (3 mg/kg), while myocardial (+)-[(3)H]PN 200-110 binding was decreased by 46-48% at 1 and 3 h later. In these rats, there was a significant decrease (34%) in cerebral cortical (+)-[(3)H]PN 200-110 binding at 3 h later. In contrast, nifedipine administration produced a similar degree of reduction (71-84%) of in vivo (+) [(3)H]PN 200-110 binding in the myocardium, aorta and cerebral cortex. It is concluded that pranidipine may exert more selective and sustained occupation in vivo of DHP receptors in vascular tissues of SHR than in myocardial and brain tissues. PMID- 10474078 TI - Sustaining effects of bopindolol on inhibitory chronotropic actions in guinea pig atria. AB - The dissociating and/or residual inhibitory effects of bopindolol from beta adrenoceptors (ARs) of atria strips pretreated with this drug and washed out with buffers on isoprenaline-induced chronotropic actions were determined. The effects of this drug were compared with those of its active metabolite 18-502, propranolol, and pindolol. Our results are as follows: (1) Lower concentrations of bopindolol (10(-9) and 10(-8) mol/l) and the active metabolite 18-502 (10(-9) mol/l), propranolol (10(-8) and 10(-6) mol/l) and pindolol (10(-8) mol/l) produced rightward shifts of concentration-response curves of isoprenaline. (2) Higher concentrations of bopindolol (10(-7) mol/l) and 18-502 (10(-8) and 10(-7) mol/l) produced a reduced maximum response by isoprenaline. (3) Bopindolol (10( 9)-10(-7) mol/l), 18-502 (10(-9)-10(-7) mol/l) and propranolol (10(-7) and 10(-6) mol/l) did not recover to control levels at 180 min even after washout with buffers. In conclusion, bopindolol and 18-502 may slowly dissociate and act as noncompetitive beta-antagonists rather than readily reversible beta-AR antagonists. These effects may differ from those of propranolol and pindolol, although propranolol did not recover to control levels after washout with buffer. PMID- 10474079 TI - Stereoselective inhibition of ethanol-induced gastric lesions in the rat by the H(3)-receptor agonist (R)-alpha-methylhistamine and its (S)-configured isomer. AB - The histamine H(3) receptor shows high degree of stereoselectivity for histamine analogues branched in the side chain. The hypothesis that gastroprotection by (R) alpha-methylhistamine could be H(3) receptor-mediated was tested by comparing the effect of (R)-alpha-methylhistamine and of (S)-alpha-methylhistamine on ethanol induced histologic lesions in the rat gastric mucosa. Extensive damage was caused by 60 min exposure to absolute ethanol, 91% of the mucosa examined being damaged. Conversely only 23% of the mucosa was damaged after pretreatment with (R)-alpha methylhistamine (100 mg/kg i.g.). In the groups pretreated with (S)-alpha methylhistamine (55.44 and 166.3 mg/kg i.g.) total damage ranged from 77 to 79%, though it was confined to the upper portion of the mucosa. Morphometric analysis of stained intraepithelial mucosubstances revealed that (R)-alpha-methylhistamine pretreatment resulted in an increase in number and volume of surface mucous cells, not evident after (S)-alpha-methylhistamine pretreatment. Scanning electron microscopy confirmed light microscopy evaluations. The two isomers of alpha-methylhistamine differently affect the response of rat gastric mucosa to absolute ethanol and they appear to differ in their influence on surface mucous cells. A basis for interpreting the effects of the two isomers of alpha methylhistamine rests on the high degree of stereoselectivity of H(3) receptors and on the different affinities of the two isomers for these receptors. PMID- 10474080 TI - Effects of mannitol and dimethylthiourea on helicobacter pylori-induced IL-8 production in gastric epithelial cells. AB - The present study aims at investigating the effects of mannitol and dimethylthiourea, known hydroxyl radical scavengers, on lipid peroxidation as an indicative of oxidative damage, NF-kappa B activation and IL-8 production by Helicobacter pylori in gastric epithelial cells. A human gastric epithelial cell line, AGS, treated with or without mannitol and dimethylthiourea, was incubated in the absence or the presence of H. pylori. As a result, H. pylori significantly stimulated the productions of lipid peroxide and IL-8. Treatment with H. pylori resulted in the activation of two species of NF-kappa B dimers (a p50/p65 heterodimer and a p50 homodimer). Mannitol and dimethylthiourea significantly inhibited lipid peroxide production, NF-kappa B complex formation and IL-8 production by H. pylori. In conclusion, mannitol and dimethylthiourea may attenuate H. pylori-induced gastric inflammation by inhibiting lipid peroxidation and NF-kappa B activation and thereby decreasing IL-8 production. PMID- 10474081 TI - Effect of wood creosote and loperamide on propulsive motility of mouse colon and small intestine. AB - To elucidate a mechanism of the antidiarrheal activity of wood creosote, its effect on the propulsive motility of mouse colon and small intestine was studied using a charcoal meal test and a colonic bead expulsion test. The effect was compared with that of loperamide. At an ordinary therapeutic dose, wood creosote inhibited the propulsive motility of colon, but not of small intestine. On the other hand, loperamide inhibited the propulsive motility of small intestine, but not of colon. The results indicate that at least a part of the antidiarrheal activity of wood creosote and loperamide is attributable to their antikinetic effect predominantly on colon of the former and predominantly on small intestine of the latter. PMID- 10474082 TI - Effects of in vivo ischemia on contractile responses of rabbit bladder to field stimulation, carbachol, ATP and KCl. AB - Rabbits were subjected to bilateral ischemia for 3, 6, or 18 h then euthanized and their bladders excised. Isolated strips of rabbit bladder detrusor were incubated in normal Tyrode's solution and contractile responses to FS, carbachol, ATP, and KCl measured. Maximal contraction, maximal rate of tension generation, and length of time to maximal contraction were determined. These studies revealed that contractile responses to FS (neurogenic stimulation) were most affected by ischemia. Contractile responses to carbachol, ATP and KCl were all similarly sensitive to ischemia. PMID- 10474083 TI - Reduction of platelet transfusion- associated sepsis by short-term bacterial culture. AB - BACKGROUND AND OBJECTIVES: There is as yet no suitable routine laboratory test for a blood transfusion service to detect bacterial contamination in platelets. This study evaluates the effectiveness and the applicability of short-term bacterial culture for such a purpose. MATERIALS AND METHODS: Samples from 5-unit platelet pools were inoculated into an aerobic culture bottle, then monitored for 48 h at 35 degrees C in an automated monitoring and detection system. RESULTS: 26,210 whole-blood-derived platelet components were tested, of which 14 (0.053%) platelet units were found to be contaminated. In addition, nine of the associated red cell units and 4 fresh-frozen plasma units grew the same organisms on culture. CONCLUSION: Short-duration bacterial culture by an automated system is effective and suitable for routine screening in a regional transfusion center. PMID- 10474084 TI - Transfusion-associated hepatitis in a tertiary referral hospital in India. A prospective study. AB - BACKGROUND AND OBJECTIVES: In Indian blood banks, screening for hepatitis B virus (HBV) is currently done by the EIA method, but no routine screening is done for hepatitis C virus (HCV). MATERIALS AND METHODS: To determine the incidence of transfusion-associated HCV hepatitis, and of any residual transfusion-associated hepatitis (TAH) after HBsAg screening, we prospectively studied 182 patients who underwent surgery and received blood transfusion. These recipients had normal alanine aminotransferase (ALT) and were negative for HBsAg (monoclonal EIA), and anti-HCV (third-generation EIA) before receiving transfusion. RESULTS: Of the 818 blood units transfused after routine screening (average 4.49+/-3.3 U/patient, range 1-14), 14 (1.7% of units) were found to be infected. Of the 182 recipients, 14 (7.69%) developed TAH during a follow-up of 6 months, 3 (21.4%) from HBV, 10 (71.5%) from HCV, and 1 (1.7%) from a coinfection of HBV and HCV. All patients with TAH due to HCV were asymptomatic. One patient with TAH due to HBV (33%) and 5 with TAH due to HCV (50%) developed chronic infection with persistently elevated ALT at 6 months. CONCLUSIONS: With the current screening practices, the incidence of TAH remains high in India and is mainly due to HCV infection. Furthermore, the screening methods for HBV also need to be improved. PMID- 10474085 TI - A randomized trial of acute normovolemic hemodilution compared to preoperative autologous blood donation in total knee arthroplasty. AB - The value of acute normovolemic hemodilution (ANH) compared to preoperative autologous blood donation (PAD) in elective surgery is controversial. We therefore conducted a prospective, randomized study to compare these techniques in patients undergoing total knee arthroplasty. ANH patients underwent up to 4 units phlebotomy or to a target hematocrit level of 28% after induction of anesthesia. PAD patients were asked to donate 1 (unilateral) or 2 (bilateral, revisions) units before admission. Mean baseline hematocrit levels were not different between ANH and PAD patients (40.6+/-4.1 vs. 38. 4+/-3.4, p = 0.09). Eight (73%) of 11 patients undergoing bilateral revision procedures received a total of 22 allogeneic blood units, whereas only 3 (14%) of 21 patients undergoing primary, unilateral procedures received a total of 3 allogeneic units (p = 0.002). We found no differences in allogeneic blood transfusions between ANH and PAD cohorts for all (n = 32) patients (1.0+/-1.2 vs. 0.6+/-1.4, p = 0.45), for unilateral knee (n = 21) replacement (0.25+/-0.46 vs. 0.08+/-0.28, p = 0.29), or for bilateral/revision (n = 11) procedures (1.9+/-1.3 vs. 2.5+/-1.9, p = 0.53). We conclude that each technique is equally effective in reducing allogeneic blood exposure. Patients undergoing revision or bilateral knee arthroplasties require adjunctive therapy to autologous blood procurement to further reduce allogeneic blood exposure. PMID- 10474086 TI - Clinical evaluation of repeat apheresis donors in Japan. AB - BACKGROUND AND OBJECTIVES: To ascertain the safety of repeat apheresis donation, hematological and biochemical tests were performed on 511 donors with a donation rate of over 6 times per year for a period of 12-19 months. MATERIALS AND METHODS: Repeat donors who had apheresis more than 6 times in the previous year were chosen. Data for the repeat donors at the start of the experiments were compared with those at the end of the study. Blood samples were taken prior to donation. Serum protein, albumin, immunoglobulin G, A, and M, serum ferritin levels were determined by biochemical tests. RESULTS: When compared to prospective donors of 400 ml, WBC, lymphocytes, and serum ferritin levels were lower in a roughly frequency-dependent manner in female and male donor groups at the beginning of the study. All the data for the male group remained almost constant with increasing frequency of apheresis donation. However, in the female group, ferritin levels significantly decreased with over 21 donations. CONCLUSIONS: The present data showed that the serum ferritin level of the female donors decreased the most with increasing frequency of apheresis donation. The cumulative RBC left in the collecting chamber and for the laboratory test is discussed in relation to a possible cause of iron deficiency in frequent apheresis donors. PMID- 10474087 TI - Adverse events in platelet apheresis donors: A multivariate analysis in a hospital-based program. AB - OBJECTIVES: This study was designed to review the incidence of adverse events during nearly 20,000 apheresis procedures over a 4-year period in a hospital based program. METHODS: Data were obtained from a review of: (1) apheresis adverse event forms (2) hospital or emergency room medical records (3) the databank for donor and procedure-related variables. Adverse events during or after the apheresis procedures were analyzed according to the following categories: (1) complications related to citrate toxicity; (2) hypotensive or vasovagal episodes; (3) complications or symptoms consistent with coronary ischemia; (4) complications related to percutaneous needle insertion, and (5) miscellaneous procedure-related events or nonspecific symptoms. Serious adverse events were categorized as persistent or severe hemodynamic changes as well as other events that required further medical evaluation. RESULTS: Of 19,736 apheresis procedures, 159 (0.81%) were associated with adverse events. In 2,376 first-time donations, 26 (1.09%) developed adverse events compared to 133 (0.77%) of 17,360 repeat procedures (p = 0.10). Seventy (0.35%) of 159 donation-related adverse events involved hemodynamic or citrate-related complications and 73 (0.37%) involved venipuncture-related complications, of which 2 required subsequent neurologic consultation. The remaining 23 (0. 12%) adverse events involved procedure-related, nonspecific complications. Forty-seven (0.24%) of the 19,736 apheresis procedures were associated with serious adverse events (SAEs). Seven of these serious adverse events required admission to an emergency department, and 2 required hospitalization for further evaluation. Multivariate analysis revealed that apheresis machine model, donor gender and weight, the concomitant harvesting of plasma, the frequency of donation, and citrate-related symptoms (e.g. paresthesias) were independently associated with severe hypotensive reactions. CONCLUSIONS: Apheresis procedures have a 150-fold higher incidence of SAEs requiring hospitalization compared to whole blood donation. Identification of donors at risk for complications can facilitate modification of the apheresis procedure in order to reduce the likelihood of adverse events. Although our study did not demonstrate a cause-effect relationship between platelet donation and the development of acute coronary syndromes, underlying cardiovascular disease was detected in 2 donors during or after the apheresis who were otherwise asymptomatic. PMID- 10474088 TI - Isoantibodies in immunoglobulin for intravenous use may cause erythrocyte sequestration. AB - BACKGROUND AND OBJECTIVES: IVIg, while generally a safe therapy for a variety of immunological disorders, can have detrimental effects on erythrocyte (RBC) homeostasis. We studied the mediation of erythrophagocytosis by isoantibodies in IVIg. MATERIALS AND METHODS: RBC were exposed to IVIg and binding of IgG determined by flow cytometry. In vitro phagocytosis of these RBC was assayed. RESULTS: Anti-A and anti-B in IVIg mediate Fc-dependent erythrophagocytosis even in the presence of excess IVIg. Removal of these isoantibodies from IVIg prevents IgG binding and erythrophagocytosis. Complement enhances IVIg-mediated RBC sequestration. CONCLUSION: It may be desirable to remove isoantibodies from IVIg especially for anemic patients or those who respond to the IVIg with a hemolytic episode. PMID- 10474089 TI - HPA genotyping by PCR-SSP: report of 4 exercises. AB - BACKGROUND AND OBJECTIVES: Polymerase chain reaction with sequence specific primers (PCR-SSP) is widely used for the determination of the alleles encoding the human platelet antigens (HPA)-1 to 5. In order to evaluate and improve performance with this technique, four exercises were organised during 1996-1998. MATERIALS AND METHODS: Coded DNA samples were distributed from the National Institute for Biological Standards and Control (NIBSC) as follows: exercise one, 18 samples; two, 12 samples; three, 6 samples, and four, 4 samples. RESULTS: Performance improved over the four exercises following the adoption of a consensus protocol and the re-design of one primer. The percentage of incorrect results in each exercise was as follows: exercise one, 9%; two 3.2%, three, 0.8%, and four, 0.3%. CONCLUSION: The modified PCR-SSP protocol is a reliable method for genotyping HPA-1 to 5 in reference laboratories. DNA-based HPA genotyping has an important role in platelet immunology and further exercises will be included in the bi-annual platelet immunology exercises organised by NIBSC. PMID- 10474090 TI - Anti-granulocyte antibody screening with extracted granulocyte antigens by a micro-mixed passive hemagglutination method. AB - BACKGROUND AND OBJECTIVES: Serologic tests for granulocyte antibodies, i.e., the granulocyte agglutination test and the granulocyte immunofluorescence test, require panels of typed granulocytes that cannot be preserved for more than a few hours. We have developed a new method in which granulocyte antigens, extracted into saline containing 3% sucrose, are coated onto U-type Terasaki plates. With this new method, we evaluated the micro-mixed passive hemagglutination test (EG MPHA) for screening for granulocyte antibodies. MATERIALS AND METHODS: We tested the ability of the EG-MPHA to detect granulocyte antigens using 5 human antibodies specific for NA1, NA2, NB1, 5b, and Sar(a), and 8 different monoclonal antibodies for NA1, CD11a, CD11b, CD13, CD16, CD18 and HLA class I. Sera from 94 alloimmunized patients were screened by the chloroquine-treated EG-MPHA method. RESULTS: NA1, NA2, NB1, 5b, Sar(a), CD11a, CD11b, CD13, CD16, CD18 and HLA class I antigens were present in the extracted granulocyte antigen preparation. CD11b and HLA class I antigens were removed when the extracted granulocyte antigens were treated with chloroquine. Granulocyte antibody screening of sera from alloimmunized patients showed that approximately 30% of the anti-HLA-positive and 10% of the anti-HLA-negative sera were positive for granulocyte antibody by the chloroquine-treated EG-MPHA. The extracted granulocyte antigen panels could be stored frozen for at least 1 year at -80 degrees C. CONCLUSION: This new method is preferable for screening for granulocyte antibodies. In addition, it has the advantage of requiring only 5 microl of serum for each test. PMID- 10474091 TI - Terminology for red cell surface antigens. ISBT Working Party Oslo Report. International Society of Blood Transfusion. PMID- 10474092 TI - Rare blood. An ISBT Working Party report on rare blood donors. International Society of Blood Transfusion. PMID- 10474093 TI - Revisiting indirect bonding PMID- 10474094 TI - A matter of priority. PMID- 10474095 TI - Soft tissue cephalometric analysis: diagnosis and treatment planning of dentofacial deformity. AB - This article will present a new soft tissue cephalometric analysis tool. This analysis may be used by the orthodontist and surgeon as an aid in diagnosis and treatment planning. The analysis is a radiographic instrument that was developed directly from the philosophy expressed in Arnett and Bergman "Facial keys to orthodontic diagnosis and treatment planning, Parts I and II" (Am J Orthop Dentofacial Orthod 1993; 103:299-312 and 395-411). The novelty of this approach, as with the "Facial Keys" articles, is an emphasis on soft tissue facial measurement. PMID- 10474096 TI - Evaluation of the consolidation period during osteodistraction using computed tomography. AB - The use of distraction osteogenesis offers an alternative approach to the correction of craniofacial deformities. However, little is known with respect to the appropriate length of the consolidation period for the newly formed bone. The objective of this study was to evaluate, by quantitative computed tomography, the regenerate bone produced during osteodistraction of the dog mandible at three different consolidation times. Twelve skeletally mature male beagle dogs were equally separated into three experimental groups. Each dog underwent 10 mm of bilateral distraction osteogenesis to lengthen the mandible. After the distraction period, the bone was allowed to consolidate for 4, 6, or 8 weeks, at which time the animals were sacrificed and the mandibles harvested for computed tomographic imaging. The results demonstrate a significantly lower mean bone density of the regenerate in the 4 week group when compared with either the 6 or 8 week groups (P < .01). There was no significant difference, however, in mean bone density between the 6 and 8 week groups. PMID- 10474097 TI - Biomechanical considerations in distraction of the osteotomized dentomaxillary complex. AB - The completely osteotomized dentomaxillary complex is essentially a free body constrained only by its soft tissue attachments. Therefore the line of action and point of application of any protractive force(s) used during distraction osteogenesis must be considered relative to its center of mass. This is in contrast to the nonsurgically separated dentomaxillary complex, which is a constrained body, and therefore the application of protractive force(s) must be considered relative to its center of resistance. These two centers are not coincident. With knowledge of the location of the center of mass, predictable protraction of the dentomaxillary complex can be achieved. In this study, the center of mass of an adult maxillary specimen osteotomized to emulate a Le Fort I osteotomy was determined. Protractive force(s) through the center of mass will produce linear advancement along its line of action. Protractive movement of the dentomaxillary complex can be adjusted downward and forward or upward and forward by locating the protractive force(s) line of action superior or inferior to the center of mass. A cleft patient is described wherein the surgically separated dentomaxillary complex is protracted downward and forward with a force vector superior to its approximate center of mass. This results in a predictable increase in overbite and overjet with negligible mandibular rotation. PMID- 10474098 TI - Comparison of frictional resistance in titanium and stainless steel brackets. AB - This study measures and compares the level of frictional resistance generated between titanium and stainless steel brackets. Both 0.018 and 0.022 inch slot size edgewise brackets were tested with different sized rectangular stainless steel wires in a specially designed apparatus. The frictional resistance was measured on Instron Universal testing machine (Instron Corp, Canton, Mass) with a 10 pound load cell. The specimen population was composed of 180 brackets and 180 wire specimens. A completely randomized design (one way) ANOVA was used to test for significant differences among the three bracket/wire types in the 0.018 and 0.022 inch slot sizes. This was followed by the Student Newman Keuls Multiple Comparison of means ranking at P < .05 to determine differences between the different groups. The titanium brackets showed lower static and kinetic frictional force as the wire size increased, whereas stainless steel brackets showed higher static and kinetic frictional force as the wire size increased. PMID- 10474099 TI - Comparison of load transmission and bracket deformation between titanium and stainless steel brackets. AB - This study measures the load transmitted and structural integrity of stainless steel and titanium brackets on application of torsional forces. Both 0.018 and 0.022 inch slot size edgewise brackets were tested in a specially designed apparatus that applied a torque value of 45 degrees. The load generated was measured by an Instron Universal Testing Machine at intervals of 15 degrees, 30 degrees, and 45 degrees of torque application. The structural stability of the brackets was evaluated by measuring the bracket slot width with a traveling stereoscopic microscope before and after the brackets were subjected to torsional forces. The specimen population was composed of 80 brackets and 80 wire specimens. An independent sample t test was used for comparison of mean load generated at 45 degrees of torque application. Repeated measures ANOVA (one-way) comparison was used to evaluate changes in load at different torque levels for the stainless steel and titanium brackets. A paired t test (two-tail) was used to determine the difference between initial and final bracket slot width at 45 degrees of torque applied for the two bracket types. The titanium brackets transmitted higher loads at 15 degrees and 30 degrees torque and lower load at 45 degrees torque on application of torsional forces in comparison to stainless steel brackets. The titanium brackets demonstrated superior dimensional stability compared to stainless steel brackets (P < .0001). PMID- 10474100 TI - Potential application of the dental aesthetic index to prioritize the orthodontic service needs in a publicly funded dental program. AB - Resources for publicly funded dental programs may never be sufficient to accommodate unlimited orthodontic service demands. There is therefore an obligation to ensure that they are preferentially provided to those patients most likely to derive the greatest benefits. To address this strategic concern, the pretreatment records of 38 patients were prioritized on a visual-analog scale relative to their differential orthodontic service needs by 16 independent orthodontic specialists. Unfortunately, inconsistencies in these assessments were unacceptable for a publicly funded program. When these same records were prioritized relative to their scores derived from the Dental Aesthetic Index, their assessments proved more consistent, especially when based on a modified Dental Aesthetic Index. The relevance of such Dental Aesthetic Index scores proved controversial, because the need for orthodontic services is dependent on other factors in addition to esthetics. The lack of established clinical guidelines to delineate the most appropriate service and timing for specific occlusal discrepancies further hampers the prioritization of orthodontic service needs. Dental Aesthetic Index case-scores may therefore be considered as just 1 of many criteria to prioritize orthodontic service needs, although the potential impact of the others has yet to be quantified. PMID- 10474101 TI - A comparison of different treatment techniques for posterior crossbite in the mixed dentition. AB - In this retrospective investigation, the changes occurring during the treatment of patients with posterior crossbite in the mixed dentition with the use of expansion plate and quad-helix appliances were evaluated and compared with those resulting from growth and development occurring in a control group of patients of similar age and type of malocclusion. The expansion plate group consisted of 13 patients, the quad-helix group of 14 patients, and the control group consisted of 10 children with transverse posterior crossbites in the mixed dentition. The research material was formed from orthodontic models and lateral and frontal cephalometric radiographs from 37 children. It was observed in this investigation that transverse expansion is achieved by both the expansion plate and quad-helix appliances. However, the average period of treatment was 1.2 years for the expansion plate, and 0.6 years for the quad-helix appliance. Although posterior crossbite was corrected in a fairly short period of time, the quad helix appliance caused considerable buccal tipping of the maxillary first permanent molars. PMID- 10474102 TI - Male adolescent facial pattern associated with TMJ disk displacement and reduction in disk length: Part II. AB - Dental practitioners continually strive to understand and determine which factors influence craniofacial morphology and how these factors may be controlled to provide the best treatment outcome for patients. Recently, an association between internal derangement of the TMJ and altered facial morphology was shown in an adolescent female sample by means of multiple regression techniques. The present study aims to determine whether similar associations are present between facial patterns exhibited by a preorthodontic male sample (n = 70) and varying degrees of TMJ internal derangement as assessed by magnetic resonance imaging. Multiple regression techniques were used to test the associations in 5 facial regions. Associations between internal derangement and craniofacial morphology differed in 3 of the facial regions when males and females were compared. However, associations between internal derangement and alteration in mandibular morphology and positioning were consistent in both gender groups. PMID- 10474103 TI - Controlled localized buckling responses of orthodontic arch wires. AB - The orthodontic arch wire is often activated locally, in transverse bending and/or longitudinal torsion, to engage an individual malaligned tooth. Arch wires with substantial flexibilities and elastic ranges in bending are available. Several clinical reports of distal displacements of molars with appliances activated by locally buckling the arch wire have appeared in the recent published literature. This article contains an explanation of buckling or "column" action and the postbuckling response of a wire, and a report of the results of a controlled, in-vitro study of a sample of 256 wire segments subjected to activation-deactivation, buckling-postbuckling-unbuckling cycles. Continuous force-displacement diagrams were obtained from mechanical tests run at oral temperature. Four orthodontics-relevant, mechanical characteristics were quantified from each diagram, and each specimen was subjected to posttest evaluation for inelastic behavior. Although the deformation of the buckled wire is, in fact, bending, the force-displacement diagrams obtained differed substantially from their familiar counterparts generated in transverse bending. Judging from the force magnitudes induced as the deactivation half-cycles commenced as well as the deactivation rates, not all of the 8 wires seem to be clinically suitable for activation initiated by buckling. Magnitudes of springback were substantial from activations as large as 6 mm, and only 2 of the 8 wires exhibited full deactivations less than 80% of their activating displacements. This relatively new mode of arch wire activation that enables delivery to the dentition of mesiodistal pushing forces has substantial potential for clinical application from several biomechanical standpoints. PMID- 10474104 TI - Severe high Angle Class II Division 1 malocclusion with vertical maxillary excess and gummy smile: a case report. AB - Severe Class II Division 1 malocclusion with vertical maxillary excess and gummy smiles can be treated in several ways. Early orthodontic treatment with vertical control may decrease the malocclusion as well as improve the appearance. In severe cases, orthognathic surgery might be the optimal solution. The following case report describes a patient with a severe gummy smile, where the final esthetic improvement was achieved by using a periodontal procedure after orthodontic treatment. PMID- 10474105 TI - Apparent hypodontia: a case of misdiagnosis. AB - The case of a 12-year-old girl is reported, whose pretreatment radiograph demonstrated agenesis of two premolars and a canine and slow development of the contralateral premolars. A follow-up radiograph taken 1 year later showed initial mineralization of a tooth germ in the site of one of the apparently missing premolars. The cause, diagnosis, and treatment planning implications of delayed mineralization and slow development of second premolars are discussed with reference to the literature. PMID- 10474106 TI - Treatment of a Class III malocclusion with maxillary constriction and an anterior functional shift. AB - This case was chosen by the CDABO student case selection committee for publication in the AJO/DO. PMID- 10474107 TI - Effect of variation in tooth morphology and bracket position on first and third order correction with preadjusted appliances. AB - Major effort is put into the design of brackets to accomplish an optimal completion of orthodontic treatment with respect to 1st, 2nd, and 3rd order corrections. Because such brackets are standardized, the intraindividual variation of the teeth is not taken into consideration. The influence of vertical bracket displacement on 1st and 3rd order corrections was studied on the plaster models, including all teeth from central incisors to first molars, of 28 young persons. The facial contours were evaluated at the mesial, central, and the distal aspect of the bracket. The contours were calculated with the formula of a parabola, and the fit was found sufficient. Intertooth variation was extreme, the biggest curvature was found among the first mandibular molars. The variation was also marked between corresponding teeth. It can be concluded that the intraindividual variation in tooth morphology is larger than the variation between the different types of preadjusted appliances. Thus, if the straight wire approach should be followed, the bracket would have to be custom made. All the calculations were made with the precondition that full size wires were used. If this is not the case, the discussion concerning individual prescription is only of limited importance. PMID- 10474108 TI - Evaluation of friction during sliding tooth movement in various bracket-arch wire combinations. AB - Frictional forces during simulated sliding tooth movement were measured with a model that was representative of the clinical condition. The model allowed tipping of the tooth until contact was established between the arch wire and diagonally opposite corners of the bracket wings; it also allowed rotation until the wire contacted opposite corners of the ligature tie, or the buccal shield with self-ligating brackets, and the base of the slot. Conventional and self ligating stainless steel brackets as well as conventional ceramic brackets, and ceramic brackets with a stainless steel slot, all with 0.022 inch bracket slot, were tested with 0.019 x 0.025 inch arch wires of stainless steel, nickel titanium, and beta titanium. Each of the 12 bracket-arch wire combinations was tested 10 times. No significant interaction was detected between brackets and arch wires (P = .89), but the bracket and arch wire effects were significant (P < .001). The pairwise differences between conventional and self-ligating stainless steel brackets and ceramic brackets with stainless steel slot were not significant. However, the conventional ceramic brackets generated significantly higher friction than the other brackets tested. Beta titanium arch wires produced higher frictional forces than nickel titanium arch wires, but no significant differences were found between each of the two and stainless steel arch wires. Attempts to identify differences in surface scratches of the arch wires produced by the different brackets were unsuccessful. PMID- 10474109 TI - Comparison of the accuracy of bracket placement between direct and indirect bonding techniques. AB - An in vitro study was conducted to evaluate the accuracy of bracket placement for direct and indirect bonding techniques. Nineteen sets of duplicated Class II malocclusion models were divided into three groups: (1) one set for ideal bracket placement, (2) nine sets for direct bonding on mannequins, and (3) nine sets for indirect bonding. Both direct and indirect bonding were performed on all teeth except molars by nine faculty members from the Department of Orthodontics, University of Pennsylvania. The position of each bonded bracket from these two bonding groups was compared with that of the same tooth from the ideal group and to each other in terms of bracket height, mesiodistal position, and angulation. Our results indicated that both direct and indirect bonding techniques failed to execute ideal bracket placement. On individual teeth, there was no statistically significant difference in the accuracy of bracket placement between these two bonding techniques except for upper right second premolar and lower left central incisor, where indirect bonding yielded better results in bracket height (P < .05), and for lower left central incisor where indirect bonding was better in mesiodistal position (P < .05), and for upper right lateral incisor where direct bonding was closer to the ideal in angulation (P < .05). Overall, indirect bonding showed better bracket placement in bracket height (P < .05), whereas, no statistically significant difference was found between them regarding the angulation and mesiodistal position. PMID- 10474111 TI - AAO COntinuing education PMID- 10474110 TI - The perceived impact of extraction and nonextraction treatments on matched samples of African American patients. AB - Orthodontic patients of African descent often seek some measure of reduction in profile protrusion. Lip retraction, however, seems to imply a need for premolar extraction. But in a nonextraction era, what does orthodontics have to offer the bimaxillary protrusion patient? To a considerable extent, it depends on what the patient wants and what treatment can provide. The present study was designed to address these questions by comparing the esthetic impact of extraction and nonextraction therapy on two morphologically similar samples of African American patients with Class I and II malocclusions. In addition to the patients, four panels of judges (black and white orthodontists and black and white laypersons) were asked to compare the pretreatment and posttreatment profile tracings and to quantify their perceptions of the esthetic impact of the change that occurred during treatment. Although there was a strong correlation among the various groups' ratings, there were a number of statistically significant differences. Premolar extraction led to a modest reduction in dental and soft tissue protrusion; nonextraction, by way of contrast, produced an increase. Presumably in response to these changes, all panels, independent of race or education, tended to prefer the posttreatment profiles, especially the posttreatment extraction profiles. There was, however, a significant interaction between race and treatment. The perceived effects of nonextraction treatment were modest and largely unrelated to the pretreatment profile, whereas the impact of premolar extraction was a highly significant function of initial protrusion: convex profiles were improved, but relatively straight profiles were made worse. The various panels, however, differed in their estimate of the point at which a profile might benefit from extraction (ie, profile reduction). For our white panelists, that point was reached when the lower lip was about 2 mm in front of Ricketts' E-plane. For our black panelists, the equilibrium point came at +4 mm. These findings imply that there is significant interaction among the type of treatment, the race of the patient, and both the race and professional background of the observer. Facial beauty, it would seem, is in the hand of the orthodontist, as well as in the eye of the beholder and the face of the beheld. PMID- 10474112 TI - The "3-D occlusogram" software. PMID- 10474113 TI - Litigation, legislation, and ethics. Degree discrimination: misleading advertising or not? PMID- 10474114 TI - Osteoprotegerin ligand and osteoprotegerin: novel implications for osteoclast biology and bone metabolism. PMID- 10474115 TI - Combined pituitary hormone deficiency - lessons from the murine models. PMID- 10474116 TI - Leukemia inhibitory factor as an intrapituitary regulator of pro-opiomelanocortin gene expression and corticotrope ontogeny. PMID- 10474117 TI - Turner syndrome. PMID- 10474118 TI - Final height in young women with Turner syndrome after GH therapy: an open controlled study. AB - GH therapy has been applied to patients with Turner syndrome for over a decade, but small sample size, delayed initiation of therapy into adolescent age and comparison with historical control subjects limit the usefulness of these studies for appraisal of the effect of GH on final adult height. We report 49 young women with Turner syndrome who completed a clinical trial in an open, non-randomized, age-matched controlled study of GH, given as daily s.c. injections at a weekly dose of 8.2 mg/m(2) for 1.9-7.5 years. Final height was defined as the measurement taken 2 years or more after height velocity declined below 2 cm/year and after a bone age of 15 'years'. The gain in height was evaluated in three ways. The mean final height gain, compared with the control group, was 4.4 cm. When corrected for the projected height at inception of therapy, the mean gained height was 5.3 cm above the control group. Shorter girls showed better response to GH then did taller girls. After correcting for parental height, the mean gain was 4.7 cm. The adult height of the GH-treated Turner women was significantly correlated with the target height, whereas no such correlation was obtained for control untreated women. Furthermore, no correlation was observed between height gain and the age or duration of GH therapy, or the age of inception of estrogen replacement therapy. It is concluded that GH therapy augments final height of girls with Turner syndrome by a mean 4.4-5.3 cm, depending on the method of evaluation, and that shorter girls may be preferred candidates for such therapy. GH therapy can be initiated after age 10 years and there is no reason to delay estrogen therapy beyond the age of 12. Indirect evidence suggests that high-dose GH therapy may surmount a pathophysiological resistance in the GH-IGF-I axis. PMID- 10474119 TI - Luteinizing hormone measurement in polycystic ovary syndrome: a practical approach. AB - The biological diagnosis of polycystic ovary syndrome (PCO) remains questionable, and a single immunological hLH (ihLH) determination can be misleading. In order better to characterize these patients, we studied hLH pulsatility every 10 min for 4h using a radioimmunoassay and then compared the results with others we obtained with a biological method. Radioimmunological and biological profiles were similar in patients with PCO and in controls. We also studied pulsatility characteristics - frequency and amplitude - and calculated the area under the curve (AUC). There was no significant increase in frequency in our 10 patients with PCO but, as in other studies, increased amplitude of hLH pulses was observed. The most discriminating parameter was the AUC. For practical purposes, we propose that hLH in patients with PCO could be assessed efficiently by taking four samples every 10 min, with computerized calculation of the AUC. PMID- 10474120 TI - Testicular adrenal rests: evidence for luteinizing hormone receptors and for distinct types of testicular nodules differing for their autonomization. AB - We report one patient with 21-hydroxylase deficiency and associated bilateral macro-orchidism caused by nodular hyperplasia of testicular adrenal rests (TAR). The boy, referred to us when 10 years old, was born with bilateral cryptorchidism that was treated unsuccessfully with i.m. injections of human chorionic gonadotropin (hCG) and later on with orchidopexy. He was treated with oral dexamethasone (0.625 mg per day) for the following 13 years. After one year, there was a marked reduction in steroid hormone levels (17-hydroxyprogesterone (17-OH P) from 27.2 to 1.2 nmol/l, testosterone from >104 to 4.8 nmol/l, estradiol (E(2)) from 481 to 33 pmol/l). After the same period of time, both testicular volume and nodularity decreased: from 45 to 18 ml and from numerous to four nodules in the right testis, and from 40 to 13 ml and from numerous to three nodules in the left testis. At the third year, there were transient increases in serum gonadotropins, testicular volume (right testis = 25 ml, left testis = 20 ml) and steroid hormones, including cortisol (serum ACTH and dehydroepiandrosterone sulfate remained suppressed). At the fourth year of follow up, there were still four nodules in the right testis and three in the left testis. The LH-dependency (which implies possession of LH/hCG receptors) of these nodules was also substantiated by their steroidogenic response to an acute i.m. hCG test. An exogenous ACTH stimulation test increased serum 17-OH P and cortisol. Since these nodules, unlike the majority of those present initially, were not suppressed by the corticosteroid therapy and since they were not detected when the patient returned for control at 23 years of age, they had partial autonomy from ACTH. At 23 years of age, the patient had a single nodule in the right testis (right testis volume = 13 ml, left testis volume = 10 ml), which should have accounted for the consistent difference in size between the two gonads. Serum LH was about 7 mU/l and FSH about 23 mU/l. The responsiveness of plasma steroid hormones to hCG had changed quantitatively and qualitatively. Secretion of cortisol was absent, secretion of 17-OH P and testosterone was reduced, and secretion of E(2) was much increased. The ACTH stimulation test showed that serum cortisol did not respond, while the other steroids responded in the order of 17-OH P>E(2)> testosterone. We conclude that there were three different groups of TAR when the patient was already 10 years old: (i) ACTH sensitive (the majority), (ii) partially ACTH-insensitive but LH/hCG-sensitive (three nodules in the left testis and three in the right testis), (iii) almost entirely ACTH-insensitive and partially hCG-insensitive (a single nodule in the right testis). Probably, the never suppressed gonadotropin levels (presumably due to the bilateral testicular damage subsequent to the cryptorchid state) and the hCG therapy were major etiological factors for the appearance of the second and third population of TAR. PMID- 10474121 TI - Apparent activities of 21-hydroxylase, 17alpha-hydroxylase and 17,20-lyase are impaired in adrenal incidentalomas. AB - OBJECTIVE: An increased response of 17-hydroxyprogesterone to ACTH stimulation has been observed in adrenal incidentaloma and linked to an impairment of either 21-hydroxylase or of 11beta-hydroxylase activity. To analyse this question further, we investigated the steroidogenic pathways in a series of 17 adrenal incidentalomas. DESIGN AND PATIENTS: 17 patients (7 women, 10 men; mean age, 62 +/- 12 years) with non-histologically analyzed adrenal incidentalomas were prospectively evaluated. METHODS: The following variables were investigated: 24-h urinary methanephrines and free cortisol excretion; plasma levels of ACTH and dehydroepiandrosterone; overnight dexamethasone suppression test; 1-24 ACTH stimulation test with measurement of: cortisol, 11-deoxycortisol, 17 hydroxyprogesterone, aldosterone, 11-deoxycorticosterone, progesterone, 17 hydroxypregnenolone, Delta4-androstenedione, dehydroepiandrosterone and 21 deoxycortisol. RESULTS: Discordant features of subclinical hypercorticism were noted in one case. No patient had dehydroepiandrosterone sulfate levels in the normal range for his or her age. Peak 17-hydroxyprogesterone and peak 21 deoxycortisol disclosed impairment of 21-hydroxylase in 11 and 10 cases respectively. An increased 11-deoxycortisol/cortisol ratio identified reduced activity of 11beta-hydroxylase in 11 patients. Eight patients displayed features of mild 17,20-lyase impairment, which was related to 21-hydroxylase dysfunction. Whereas only 2 patients showed no enzyme modification, 9 displayed alterations of at least two pathways. CONCLUSION: In our hands, a combination of enzyme dysfunction was frequently observed. Shared biochemical mechanisms could explain combined 17,20-lyase and 21-hydroxylase alterations, whereas coexistence of 21 hydroxylase (particularly when based on peak 21-deoxycortisol) and 11beta hydroxylase is more puzzling. PMID- 10474122 TI - LH and FSH secretory responses to GnRH in normal individuals: a non-parametric deconvolution approach. AB - OBJECTIVE: To reconstruct the instantaneous secretion rate (ISR) of LH and FSH after GnRH administration in normal volunteers using non-parametric deconvolution, and to derive a direct integration formula to evaluate the amount of LH and FSH secreted during the first 60 min after the stimulus. DESIGN AND METHODS: First, the deconvolution method was validated in vivo by reconstructing doses ranging from 7.5 IU to 75 IU injected in three healthy adult volunteers whose endogenous LH had previously been downregulated by pretreating them, 3-4 weeks earlier, with 3.75 mg GnRH agonist i.m. Then, 40 healthy adult male volunteers were tested with a single 100 microg GnRH bolus, administered at 0 min. LH and FSH concentrations were determined at -30, 0, 15, 30, 45, 60, 90, and 120 min. RESULTS AND CONCLUSIONS: The validation study, conducted over a 10-fold range of doses, demonstrated that non-parametric deconvolution provided a reasonably accurate estimate of the amount of hormone entering the circulation. Applying deconvolution to the LH and FSH responses to GnRH, the ISRs of both hormones were shown to have a similar pattern, with a clearly delimited pulse after the GnRH bolus. In conjunction with earlier analyses of estimates of GHRH stimulated GH secretion, we conclude that secretagogues evoke discrete LH, FSH, and GH secretory bursts of about 60 min total duration, despite markedly unequal (glyco-)protein hormone half-lives (18-500 min). With respect to the assessment of total hormone release during the first 60 min after the stimulus, the integration formula provided a reliable approximation of the result obtained by deconvolution, and had a negligible dependence on the samples at times 90 and 120 min. PMID- 10474123 TI - Two-week pulsatile gonadotropin releasing hormone infusion unmasks dual (hypothalamic and Leydig cell) defects in the healthy aging male gonadotropic axis. AB - OBJECTIVE: To examine the possibility that lower serum bioavailable testosterone concentrations, without increased LH release, in healthy older men, reflects hypothalamic GnRH deficiency. DESIGN: We used a randomized, double-blind, placebo controlled design. METHODS: We treated each of five young (ages 20-34 years) and five older (ages 60-78 years) men with 2 weeks of randomized infusions of saline or pulsatile GnRH (100 ng/kg i.v. every 90 min). RESULTS: At baseline (saline infusion), older men had more LH pulses (young compared with old, 10 +/- 0.6 compared with 15 +/- 1, P = 0.0026) per 24h, reduced fractional LH pulse amplitude (219 +/- 17% compared with 167 +/- 40%, P = 0.0376), and more disorderly hormone release as judged by approximate entropy (ApEn) (LH, P < or = 0.0001; testosterone, P < or = 0.0047). In response to pulsatile i.v. GnRH infusions, serum 24-h LH concentrations (measured by immunoradiometric assay (IRMA)), increased equivalently in young and older men (to 7.3 +/- 1.2 and 7.2 +/ 1.8 IU/l respectively). GnRH treatment also normalized LH pulse frequency and amplitude, ApEn, and plasma biologically active LH (pooled) concentrations. In contrast, 24-h testosterone concentrations failed to increase equivalently in older men (young compared with old, 869 +/- 88 compared with 517 +/- 38 ng/dl, P = 0.0061), reflecting lower testosterone peak maxima (995 +/- 108 compared with 583 +/- 48 ng/dl, P = 0.0083) and interpeak nadirs (750 +/- 87 compared with 427 +/- 26 ng/dl, P = 0.0073). CONCLUSIONS: We have demonstrated that, in older men, successful reconstitution of 24-h pituitary (bioactive) LH output and pulsatile (IRMA) LH release patterns could be achieved by a fixed exogenous GnRH pulse signal, thereby implicating altered endogenous hypothalamic GnRH release in the relative hypogonadotropism of aging. The failure of testosterone concentrations to increase concomitantly points to a simultaneous Leydig cell defect. We conclude that aging in men is marked by a dual defect in the central nervous system-pituitary-Leydig cell axis. PMID- 10474124 TI - A comparison between octreotide-LAR and lanreotide-SR in the chronic treatment of acromegaly. AB - BACKGROUND: At present long-acting somatostatin analogs represent the first-line medical treatment of acromegaly. These drugs produce stable suppression of GH in most sensitive patients and IGF-I normalization in many; they also increase the compliance of acromegalic patients. The recent availability of octreotide (OC) LAR, a somatostatin analog to be administered at 28-day intervals, has prompted us to compare, in the same group of patients, its effects and those of another somatostatin analog already available, lanreotide-SR (LSR, to be administered at 14-day intervals). PATIENTS: Twelve somatostatin analog-sensitive acromegalic patients with active disease were enrolled in a prospective open sequential study after giving their informed consent. After chronic treatment with LSR (6-24 months), the patients were changed to treatment with OC-LAR, without wash-out. LSR had been administered at individually tailored dosages (30 mg i.m. at 7-21 day intervals, median 10 days - every 7 days in seven patients, 10 days in two patients, 14 days in two patients and 21 days in one patient) according to GH and IGF-I responses. Disease stability was obtained, as shown by maximal GH/IGF-I suppression without any significant hormonal change between the last two control measurements. OC-LAR was administered i.m. at 28-day intervals six times at the dosage of 20 mg for the first three times and 10 or 30 mg for the last three times (according to individual GH/IGF-I responses). GH (mean of three, hourly samples) and IGF-I concentrations were evaluated on the same day as each administration of the drug, before its injection. RESULTS: GH and IGF-I values were significantly decreased by LSR treatment. GH decreased from 41.6 +/- 14.6 microg/l (mean +/- s.e.) to 7.2 +/- 1.5 microg/l (P < 0.02), whereas IGF-I values declined from 959 +/- 95 microg/l to 460 +/- 61 microg/l (P < 0.00001), expressed as absolute values, and from 287 +/- 30% to 137 +/- 19% expressed as percentage of the upper limit of normal range (% ULNR). At the end of the last cycle, OC-LAR treatment achieved a significant further suppression both in GH (to 5.1 +/- 1.1 microg/l, P < 0.05 compared with LSR) and in IGF-I concentrations (to 374 +/- 60 microg/l, P<0.05 compared with LSR, and to 112 +/- 19% as % ULNR). LSR decreased GH concentrations to less than 2.5 microg/l in one patient and normalized IGF-I concentrations in four patients. OC-LAR decreased GH concentrations to less than 2.5 microg/l in four patients and normalized or near-normalized IGF-I concentrations (i.e. to < 110% ULNR) in eight patients. CONCLUSIONS: These preliminary results show that the once-monthly OC-LAR administration schedule proved more efficacious than LSR given every 7-21 days, in a greater number of acromegalic patients. PMID- 10474125 TI - T-cell mediated autoimmunity to the insulinoma-associated protein 2 islet tyrosine phosphatase in type 1 diabetes mellitus. AB - The target molecules of the T-cell response in type 1 diabetes, despite their pathogenic importance, remain largely uncharacterized, especially in humans. Interestingly, molecules such as insulin and glutamic acid decarboxylase (GAD) have been shown to be a target not only of autoantibodies, but also of autoreactive T-lymphocytes both in man and in the non-obese diabetic (NOD) mouse. In the present study we aimed to determine the existence of a specific T-cell response towards the insulinoma-associated protein 2 (IA-2) islet tyrosine phosphatase, a recently identified autoantigen which is the target of autoantibodies strongly associated with diabetes development. Human recombinant IA-2 produced in Escherichia coli, was tested for its reactivity with peripheral blood lymphocytes obtained from 16 newly diagnosed type 1 diabetic patients and from 25 normal controls, 15 of whom were HLA-DR-matched. A T-cell proliferation assay was performed in triplicate employing freshly isolated cells in the absence or in the presence of the antigen to be tested (at two different concentrations: 2 microg/ml and 10 microg/ml). A specific T-cell proliferation (defined as a stimulation index (S.I.) >/=3) was observed against IA-2 used at a concentration of 10 microg/ml (but not of 2 microg/ml) in 8/16 diabetic patients, in 1/15 HLA DR-matched control subjects (P<0.01 by Fisher exact test) and in 0/10 of the remaining normal individuals. A statistically significant difference (P<0.003 by Mann-Whitney U test) was also observed in S.I. values between patients (3.1+/ 1.4) and HLA-DR-matched controls (1.7+/-0.54) employing IA-2 at a concentration of 10 microg/ml. However, when IA-2 was used at a concentration of 2 microg/ml, the difference in S. I. between patients (1.65+/-0.8) and controls (1.0+/-0.3) did not reach statistical significance. In conclusion, these data show the presence of a specific, dose-dependent T-lymphocyte response against the IA-2 islet tyrosine phosphatase at the onset of type 1 diabetes. Consequently, this molecule appears to be a target not only at the B-lymphocyte but also at the T lymphocyte level, reinforcing the potential pathogenic role of this autoantigen in the islet destructive process. PMID- 10474126 TI - Aldosterone-producing adenoma without hypertension: a report of two cases. AB - Normotensive primary hyperaldosteronism is exceedingly rare. We report two new cases of this syndrome in two middle-aged women, one of Asian origin. The presenting signs were tetany in one case and an adrenal mass in the other. Neither patient had hypertension, despite repeated measurements with a manual armlet. A typical biological profile of primary hyperaldosteronism was demonstrated in both patients, including hypokalemia with inappropriate kaliuresis, elevated resting plasma aldosterone, and undetectable plasma renin activity. The circadian rhythm of blood pressure was studied by ambulatory monitoring pre- and post-operatively. It confirmed the lack of hypertension, but the circadian rhythm of blood pressure was lost before surgery in one patient. Surgical removal of the histologically typical aldosterone-producing adenomas normalized the kalemia. The main finding in these two patients was spontaneously low blood pressure in the post-operative period. This suggests that excess aldosterone induced relative hypertension in these patients whose blood pressure was spontaneously very low. Genetic screening for dexamethasone-sensitive hyperaldosteronism was negative in both patients. PMID- 10474127 TI - Incidental detection of familial medullary thyroid carcinoma by calcitonin screening for nodular thyroid disease. AB - Serum calcitonin screening has recently been found to be a useful supplement to fine-needle aspiration biopsy, ultrasound and radionuclide imaging in the evaluation of thyroid nodules. We describe a case where introduction of routine calcitonin screening in nodular thyroid disease led to the detection of a family with medullary thyroid carcinoma. The benefits and problems of basal and stimulated serum calcitonin testing and ret-proto-oncogene mutation studies are exemplified and we discuss the appropriate use and interpretation of these tests. We conclude that routine basal serum calcitonin measurement in nodular thyroid disease and thoughtful use of ret-mutation analysis is cost-effective in detecting medullary thyroid carcinoma and multiple endocrine neoplasia type II. PMID- 10474128 TI - Abnormalities of hypothalamic-pituitary-adrenal and hypothalamic-somatotrophic axes in Fawn-Hooded rats. AB - Fawn-Hooded (FH) rats show central and peripheral abnormalities in serotoninergic functions and have attracted attention as an animal model of some pathologies, including depression and hypertension. In addition, these rats show a reduced growth rate. As the hypothalamic-pituitary-adrenal (HPA) axis has been implicated in both depression and hypertension, and the hypothalamic-somatotrophic (HSM) axis has a major role in growth, these two endocrine axes were characterised in FH rats as compared with outbred Sprague-Dawley (SD) rats in basal conditions. FH rats showed normal serum ACTH and corticosterone concentrations, but reduced serum corticosterone binding capacity. At a central level, normal expression of mRNA for glucocorticoid type II receptors in the hippocampal formation and mRNA for corticotrophin-releasing factor (CRF) in the paraventricular nucleus of the hypothalamus were observed in FH rats, whereas expression of mRNA for CRF in the central nucleus of the amygdala was enhanced compared with the expression in SD rats. Serum GH concentrations were normal in FH rats, IGF-I tended to be lower, and mRNA for somatostatin (SRIF) in the periventricular nucleus of the hypothalamus was significantly lower in FH rats than in SD rats. The reduced SRIF gene expression in rats with normal or slightly reduced GH and IGF-I, respectively, might be secondary to a defective central and peripheral response to IGF-I, compatible with the reduced growth of FH rats. The present results suggest that FH rats have abnormalities in both HPA and HSM axes that might be related to some of their physiopathological characteristics. PMID- 10474129 TI - The effect of combined oestrogen and progesterone replacement on the renal responses to oxytocin and vasopressin in ovariectomized rats. AB - OBJECTIVE: Renal responsiveness to the neurohypophyseal hormones, oxytocin and vasopressin, has been shown in the rat to vary during pregnancy and lactation. A study was performed to determine whether ovarian steroids could contribute to the observed changes. DESIGN: Using a previously validated method, fluid excretion during infusion of oxytocin or vasopressin was monitored in ovariectomized animals with and without chronic administration of oestrogen and progesterone. METHODS: After 14 days treatment with vehicle or 12.5 mg hydroxyprogesterone caproate and 0.25 mg oestradiol valerate injected every 3 days, rats were infused with 0.077 mol/l NaCl for an equilibration period of approximately 2.5h. Timed urine collections for the determination of volume and electrolytes were then made during a control period of at least 45 min and for 60 min while the infusate was supplemented with vasopressin (40 fmol/min) or oxytocin (50 fmol/min). Further observations were made for a final 90 min of hypotonic saline infusion. In control infusions saline alone was given. RESULTS: Treatment with ovarian steroids did not affect the volume of urine excreted during hormone infusion. Electrolyte excretion, however, was affected with lower concentrations of sodium and chloride on oxytocin infusion being seen in the steroid-treated animals. During vasopressin infusion, peak electrolyte concentrations were also achieved later in this group of animals. CONCLUSION: The increased circulating concentrations of oestrogen and progesterone seen during pregnancy could contribute to variations in the natriuretic response to neurohypophyseal hormones observed in the rat. PMID- 10474130 TI - Continuous s.c. infusion rather than twice-daily injections of IGF-I more effectively increases serum IGF binding protein-3 in female monkeys. AB - OBJECTIVE: In order to better understand how the IGF-I axis is affected by exogenous IGF-I, this study compared the effects of a constant s.c. infusion of IGF-I with that of twice-daily injections of IGF-I in young adult female rhesus monkeys. Clinical studies suggest that circulating concentrations of insulin-like growth factor binding protein-3 (IGFBP-3) are decreased or unaffected by IGF-I administration, whereas acute increases in IGF-I may increase serum IGFBP-1. However, studies in monkeys indicate that acute or continuous infusion of IGF-I effectively increases serum IGFBP-3. DESIGN AND METHODS: Female monkeys were studied for 5 days with no IGF-I supplementation (baseline) and for 5 days of IGF I treatment by either constant infusion (120 microg/kg per day s.c., n = 5) or twice-daily injections of IGF-I (60 microg/kg per injection s.c., n = 5). Serum samples were collected daily at 0800 h and at 0800, 0900, 1100, 1500, and 2000 h on days 1 and 4 for each condition. Samples were assayed for IGF-I, IGFBPs-1 and 3, insulin, and glucose. RESULTS: Serum IGF-I was consistently increased above baseline within 24 h of the initiation of constant infusion, but was delayed until the second day of treatment in the injection group. Serum IGFBP-3 followed the pattern of IGF-I, with concentrations increased by day 1 during constant infusion and by day 2 during intermittent injections. Although both treatments effectively increased serum IGFBP-3, the increase was greater during constant infusion (31% above baseline) compared with injection (17%). Immunoblotting revealed that the constant infusion of IGF-I resulted in quantitatively more lower-molecular-mass fragments of IGFBP-3 than were observed during baseline or intermittent injections. Size-exclusion chromatography and ultrafiltration indicated that most IGFBP-3 was found in the ternary complex, with a greater percentage found in the ternary complex during baseline (90%) than during constant infusion (86%) or intermittent injections of IGF-I (87%). In contrast, serum concentrations of IGFBP-1 were increased on day 1 of both treatments, but declined towards baseline values as treatment progressed. Serum concentrations of insulin and glucose were unaffected by either mode of IGF-I treatment. Serum concentrations of IGF-I and IGFBP-3 were increased within 3h of the injection, before declining towards the pre-injection level. In contrast, the daily pattern of serum hormone concentrations was similar between the baseline condition and during constant infusion of IGF-I. Although higher during the treatment phase, serum IGF-I and IGFBP-3 concentrations decreased significantly from 0800 h until the afternoon meal, reaching a nadir in the evening before increasing again the next morning. Serum insulin decreased also after the morning meal and increased significantly immediately after the afternoon meal. Although serum IGFBP-1 also decreased initially after the morning meal, concentrations reached a peak before the afternoon meal as serum insulin reached its nadir. CONCLUSION: The results of the present analysis indicate that the constant infusion of IGF-I more effectively sustains serum concentrations of IGF-I and IGFBP-3 than do twice daily injections. Although the percentage of IGF-I and IGFBP-3 in the ternary complex was similar during both treatments, the constant infusion regimen produced lower-molecular-mass fragments of IGFBP-3. In addition, serum IGF-I and IGFBP-3 appeared to be regulated diurnally, even during IGF-I infusion, whereas IGFBP-1 and insulin were affected by the timing of food intake. Taken together, these data suggest that, in the monkey, IGFBP-3 is regulated by factors in addition to GH, and that IGF-I can affect its own bioavailability by increasing circulating concentrations of IGFBP-3. PMID- 10474132 TI - Preface. PMID- 10474131 TI - Six-week treatment with hexarelin in young dogs: evaluation of the GH responsiveness to acute hexarelin or GHRH administration, and of the orexigenic effect of hexarelin. AB - In this study we evaluated, in six young (5-7 year-old) beagle dogs, the effects of a 6-week administration of hexarelin (250 microg/kg s. c. twice daily) on the GH response to an acute challenge with hexarelin or GHRH (2 microg/kg i.v.), delivered before and after 3 and 6 weeks of treatment. The GH peak response to acute hexarelin or GHRH initially increased, with a maximum observed at the 3rd week, and then decreased to basal values (GHRH) or less (hexarelin) at the 6th week. These data would indicate that hexarelin initially primed the pituitary to acute administration of further hexarelin or of GHRH, followed by downregulation of the GH response to hexarelin and preservation of the response to GHRH. We then studied the rebound increase in GH secretion after withdrawal of an infusion of somatostatin (4 microg/kg per h for 1.5 h), a likely stimulus of endogenous GHRH function. The pattern obtained was similar to, though not superimposable upon, that ensuing after acute hexarelin or GHRH administration. Parallel evaluation of the acute orexigenic effect of hexarelin evinced a different time-course of the behavioural response, namely an acute feeding response to hexarelin that was abolished at the 3rd week and returned to normal at the 6th week. The differing timing of the neuroendocrine or behavioural response to hexarelin would suggest the existence of different subtypes of central nervous system GH-releasing peptide receptors. PMID- 10474133 TI - Modelling ordered categorical data: recent advances and future challenges. AB - This article summarizes recent advances in the modelling of ordered categorical (ordinal) response variables. We begin by reviewing some models for ordinal data introduced in the literature in the past 25 years. We then survey recent extensions of these models and related methodology for special types of applications, such as for repeated measurement and other forms of clustering. We also survey other aspects of ordinal modelling, such as small-sample analyses, power and sample size considerations, and availability of software. Throughout, we suggest problem areas for future research and we highlight challenges for statisticians who deal with ordinal data. PMID- 10474134 TI - Some practical issues in binary data analysis. AB - Three topics motivated by practical problems where the response variable is binary are described and illustrated. When a number of different explanatory variables are measured on each individual, a parsimonious model may be needed to predict the response of a future patient, or in selecting the variables that any treatment effect must be adjusted for. Some variable selection procedures used in conjunction with fitting logistic regression models are summarized and their performance investigated using a simulation study. A study to compare two devices for delivering anaesthetic gas to patients during surgery is then described, in which the response variable is the incidence of post-operative sore throat. In this study, the allocation of patient to device was non-random and a method for analysing these data that takes account of this aspect of the data is illustrated. In studies to compare different forms of contraceptive, the extent of regularity in the menstrual bleeding cycle is an important consideration for the acceptability of a contraceptive. Diary data on the menstrual bleeding pattern are therefore routinely collected. A method of summarizing the cyclic behaviour in the diary data for a particular woman is described, and extended to allow comparisons to be made between groups of women on different types of contraceptive. The approach is illustrated using a database made available by the World Health Organization. PMID- 10474135 TI - A review of some extensions to generalized linear models. AB - Although generalized linear models are reasonably well known, they are not as widely used in medical statistics as might be appropriate, with the exception of logistic, log-linear, and some survival models. At the same time, the generalized linear modelling methodology is decidedly outdated in that more powerful methods, involving wider classes of distributions, non-linear regression, censoring and dependence among responses, are required. Limitations of the generalized linear modelling approach include the need for the iterated weighted least squares (IWLS) procedure for estimation and deviances for inferences; these restrict the class of models that can be used and do not allow direct comparisons among models from different distributions. Powerful non-linear optimization routines are now available and comparisons can more fruitfully be made using the complete likelihood function. The link function is an artefact, necessary for IWLS to function with linear models, but that disappears once the class is extended to truly non-linear models. Restricting comparisons of responses under different treatments to differences in means can be extremely misleading if the shape of the distribution is changing. This may involve changes in dispersion, or of other shape-related parameters such as the skewness in a stable distribution, with the treatments or covariates. Any exact likelihood function, defined as the probability of the observed data, takes into account the fact that all observable data are interval censored, thus directly encompassing the various types of censoring possible with duration-type data. In most situations this can now be as easily used as the traditional approximate likelihood based on densities. Finally, methods are required for incorporating dependencies among responses in models including conditioning on previous history and on random effects. One important procedure for constructing such likelihoods is based on Kalman filtering. PMID- 10474136 TI - Marginal modelling of multivariate categorical data. AB - This paper describes likelihood methods of analysis for multivariate categorical data. The joint distribution is specified in terms of marginal mean functions, and pairwise and higher order association measures. For the association, the emphasis is on global odds ratios. The method allows flexible formulation of a broad class of designs, such as repeated measurements, longitudinal studies, interrater agreement and cross-over trials. The proposed model can be used for parameter estimation and hypothesis testing. Simple fitting algorithms are proposed. The method is illustrated using a data example. PMID- 10474137 TI - Dispersion models and longitudinal data analysis. AB - Dispersion models provide a flexible class of non-normal distributions with many potential applications in biostatistics, accommodating a wide range of continuous, discrete and mixed data. Starting with Liang and Zeger's generalized estimating equation method, we review some recent applications of dispersion models in longitudinal data analysis, including state space models based on the Tweedie class of exponential dispersion models. In medical applications the latent process of a state space model may often be interpreted as an unobserved potential morbidity process, which is modelled as a function of time varying covariates. By allowing a multivariate response vector of 'symptoms', the model integrates several response variables of mixed types into a single model. For growth curve models, the latent process reflects the 'true' growth. PMID- 10474138 TI - A unified theory for sequential clinical trials. AB - The theory underlying sequential clinical trials is now well developed, and the methodology is increasingly being implemented in practice, both by the pharmaceutical industry and in the public sector. The consequences of conducting interim analyses for frequentist interpretations of data are now well understood. A large number of approaches are available for the calculation of stopping boundaries and for the eventual terminal analysis. In this paper, the principles of the design and analysis of sequential clinical trials will be presented. Existing methods will be reviewed, and their relationships with the general principles will be clarified. Controversies and gaps within the methodology will be highlighted. It is intended that presentation of the subject as a single unified theory will allow the few essential underlying features to be better appreciated. PMID- 10474139 TI - Group sequential analyses for the mean function of a repeated measure process. AB - In many clinical trials, treatment efficacy is based upon response to a biological marker that is measured repeatedly during the course of follow-up. However, in some of these trials it is not clear, a priori, how treatment effects on the marker may manifest themselves or what kinds of effects are clinically meaningful and/or acceptable. It is, therefore, desirable to allow flexibility in design and monitoring process by not prespecifying a stopping rule or even the parameter on which inferences will be based. Using the more general results in Hu and Lagakos, this paper extends the idea of the repeated confidence intervals for a parameter (Jennison and Turnbull) to repeated confidence bands for the mean function of a repeated measure process. We illustrate the approach and some considerations in its application with the results of a recent AIDS clinical trial. PMID- 10474140 TI - Emerging and recurrent issues in drug development. AB - This paper reviews several emerging and recurrent issues relating to the drug development process. These emerging issues include changes to the FDA regulatory environment, internationalization of drug development, advances in computer technology and visualization tools, and efforts to incorporate meta-analysis methodology. Recurrent issues include: renewed interest in statistical methods for handling subgroups in the design and analysis of clinical trials; renewed interest in alternatives to the 'intention-to-treat' analysis in the presence of non-compliance in randomized clinical trials; renewed interest in methodology to address the multiplicities resulting from a variety of sources inherent in the drug development process, and renewed interest in methods to assure data integrity. These emerging and recurrent issues provide a continuing challenge to the international community of statisticians involved in drug development. Moreover, the involvement of statisticians with different perspectives continues to enrich the field and contributes to improvement in the public health. PMID- 10474141 TI - Statistics versus statistical science in the regulatory process. AB - This paper reviews the established practice of providing evidence to regulatory authorities about the claimed properties (such as efficacy and safety) of new pharmaceutical products. The established conventions and procedures are contrasted with scientific concepts and principles. The following issues are discussed: (a) recruitment of subjects and its connection to treatment heterogeneity; (b) the measurement process and the handling of missing data; (c) data transformation and the use of generalized linear models; (d) model selection and model checking; (e) the 'cult of the single trial' and the use of prior information; and (f) hypothesis testing and the P-value culture. PMID- 10474142 TI - Controversies about sponsor initiated re-analyses of clinical trial data in the licensing process. AB - I present my experience with sponsor initiated re-analyses as a response to the regulatory agency's 'Mangelbericht', and the difficulties which arose from the fact that the regulatory agency was not involved in the plans for correcting mistakes or irregularities in the data and for the (re-)analysis of them. I also discuss the related problem when there are major discrepancies between the planned procedures laid down in the protocol and the actually applied procedures in the study report. I give examples for poor planning of a clinical study (sample size, statistical analysis, target variable(s)) and how this reduces the strength and value of a study. I introduce the phrase 'neutral party' as a means to resolve regulatory concerns about partiality in the sponsor's decisions, for example, introduction of new statistical models, different from the one planned, change in the order of the target variables, or dropping variables from the list of target variables. PMID- 10474143 TI - Diagnostic statistical procedures in medical meta-analyses. AB - The number of published meta-analyses in medicine has had phenomenal growth, to a point where over 300 meta-analyses in medicine are published yearly. Because meta analyses tend to lead to policy decisions, it is extremely important that the analyses be robust and that alternative analyses yield consistent results. We herein provide a discussion of diagnostic statistical procedures. PMID- 10474144 TI - Meta-analysis by random effect modelling in generalized linear models. AB - The meta-analysis of multi-centre trials can be based on either fixed or random effect models. This paper argues for the general use of random effect models, and illustrates the value of non-parametric maximum likelihood (NPML) analysis of such trials. The same general approach unifies administrative 'league table' analyses in epidemiological and other studies. Several examples of the NPML analysis are given, including a 70-centre trial. PMID- 10474145 TI - Event history analysis and inference from observational epidemiology. AB - Systematic inclusion of time in observational epidemiological studies may help strengthen the inference to be drawn, but new epidemiological challenges arise, such as time-dependent confounders - covariates which may change from being confounders to being intermediate variables. The focus of this presentation concerns two sets of tools: event history analysis and structural nested failure time models, both applied to a particularly intricate problem in observational epidemiology, of empirically assessing the graft-versus-leukaemia effect after bone marrow transplantation. PMID- 10474146 TI - State duration models in clinical and observational studies. AB - Many studies in medicine involve conditions whereby subjects make transitions among a set of defined states over time. In such situations the durations of sojourns in specific states is frequently of interest. This article considers the modelling and analysis of sojourn times, beginning with semi-Markov models in which the durations of different sojourns are independent, and then considering extended models incorporating chronological time effects and random effects. Methodologic challenges for inference are discussed, and examples involving a relapse-remitting process and recurrent events are considered. PMID- 10474147 TI - Issues in the statistical analysis of small area health data. AB - The availability of geographically indexed health and population data, with advances in computing, geographical information systems and statistical methodology, have opened the way for serious exploration of small area health statistics based on routine data. Such analyses may be used to address specific questions concerning health in relation to sources of pollution, to investigate clustering of disease or for hypothesis generation. We distinguish four types of analysis: disease mapping; geographic correlation studies; the assessment of risk in relation to a prespecified point or line source, and cluster detection and disease clustering. A general framework for the statistical analysis of small area studies will be considered. This framework assumes that populations at risk arise from inhomogeneous Poisson processes. Disease cases are then realizations of a thinned Poisson process where the risk of disease depends on the characteristics of the person, time and spatial location. Difficulties of analysis and interpretation due to data inaccuracies and aggregation will be addressed with particular reference to ecological bias and confounding. The use of errors-in-variables modelling in small area analyses will be discussed. PMID- 10474148 TI - Statistical thinking in functional and structural magnetic resonance neuroimaging. AB - Burning issues abound in the emerging field of biostatistics for imaging studies of the living human brain. This paper employs two examples to discuss the roles of informed empirical models and modern statistical methods in the analysis of functional and structural magnetic resonance neuroimages. The first example derives from an analysis of a functional baseline study in which simulated signals are embedded in actual brain 'noise' generated by a resting subject lying passively in the imaging device. The second example is taken from a structural volumetric analysis of human brain structure size differences in a sample of healthy adults. PMID- 10474149 TI - Analysis of recurrent events: application to a clinical trial of colony stimulating factor with the endpoint of febrile neutropenia. AB - This paper discusses the analysis of randomized clinical trials where the primary endpoint is a recurrent event observed during a time period which can vary among patients. In particular it describes a method of estimating robust confidence limits for clinically relevant parameters. As an illustrative example, a clinical trial of a macrophage colony stimulating factor (M-CSF) for patients with acute myeloblastic leukaemia (AML) is introduced where the endpoint is the incidence and duration of febrile neutropenia. The causal effects in the framework of Rubin's deterministic model are defined as estimands. A non-parametric method based on the permutation test, using an accelerated search procedure proposed by Garthwaite, is compared with other standard methods by simulation and by application to the AML trial data. PMID- 10474150 TI - Ignorability and bias in clinical trials. AB - Patient non-compliance and drop-out can bias analyses of clinical trial data. I describe a parametric model for treatment cross-over and drop-out and demonstrate how the concept of ignorability, originally defined for incomplete-data problems, can elucidate sources of bias in clinical trials. I discuss some implications of the theory and present simulation examples that illustrate the potential effects of non-ignorable cross-over and drop-out on bias and power. PMID- 10474151 TI - Non-ignorable missing covariates in generalized linear models. AB - We propose a likelihood method for estimating parameters in generalized linear models with missing covariates and a non-ignorable missing data mechanism. In this paper, we focus on one missing covariate. We use a logistic model for the probability that the covariate is missing, and allow this probability to depend on the incomplete covariate. We allow the covariates, including the incomplete covariate, to be either categorical or continuous. We propose an EM algorithm in this case. For a missing categorical covariate, we derive a closed form expression for the E- and M-steps of the EM algorithm for obtaining the maximum likelihood estimates (MLEs). For a missing continuous covariate, we use a Monte Carlo version of the EM algorithm to obtain the MLEs via the Gibbs sampler. The methodology is illustrated using an example from a breast cancer clinical trial in which time to disease progression is the outcome, and the incomplete covariate is a quality of life physical well-being score taken after the start of therapy. This score may be missing because the patients are sicker, so this covariate could be non-ignorably missing. PMID- 10474152 TI - Missing data perspectives of the fluvoxamine data set: a review. AB - Fitting models to incomplete categorical data requires more care than fitting models to the complete data counterparts, not only in the setting of missing data that are non-randomly missing, but even in the familiar missing at random setting. Various aspects of this point of view have been considered in the literature. We review it using data from a multi-centre trial on the relief of psychiatric symptoms. First, it is shown how the usual expected information matrix (referred to as naive information) is biased even under a missing at random mechanism. Second, issues that arise under non-random missingness assumptions are illustrated. It is argued that at least some of these problems can be avoided using contextual information. PMID- 10474153 TI - On summary measures analysis of the linear mixed effects model for repeated measures when data are not missing completely at random. AB - Subjects often drop out of longitudinal studies prematurely, yielding unbalanced data with unequal numbers of measures for each subject. A simple and convenient approach to analysis is to develop summary measures for each individual and then regress the summary measures on between-subject covariates. We examine properties of this approach in the context of the linear mixed effects model when the data are not missing completely at random, in the sense that drop-out depends on the values of the repeated measures after conditioning on fixed covariates. The approach is compared with likelihood-based approaches that model the vector of repeated measures for each individual. Methods are compared by simulation for the case where repeated measures over time are linear and can be summarized by a slope and intercept for each individual. Our simulations suggest that summary measures analysis based on the slopes alone is comparable to full maximum likelihood when the data are missing completely at random but is markedly inferior when the data are not missing completely at random. Analysis discarding the incomplete cases is even worse, with large biases and very poor confidence coverage. PMID- 10474154 TI - Multivariate linear mixed models for multiple outcomes. AB - We propose a multivariate linear mixed (MLMM) for the analysis of multiple outcomes, which generalizes the latent variable model of Sammel and Ryan. The proposed model assumes a flexible correlation structure among the multiple outcomes, and allows a global test of the impact of exposure across outcomes. In contrast to the Sammel-Ryan model, the MLMM separates the mean and correlation parameters so that the mean estimation will remain reasonably robust even if the correlation is misspecified. The model is applied to birth defects data, where continuous data on the size of infants who were exposed to anticonvulsant medications in utero are compared to controls. PMID- 10474155 TI - Innovations in bayes and empirical bayes methods: estimating parameters, populations and ranks. AB - By formalizing the relation among components and 'borrowing information' among them, Bayes and empirical Bayes methods can produce more valid, efficient and informative statistical evaluations than those based on traditional methods. In addition, Bayesian structuring of complicated models and goals guides development of appropriate statistical approaches and generates summaries which properly account for sampling and modelling uncertainty. Computing innovations enable implementation of complex and relevant models, thereby substantially increasing the role of Bayes/empirical Bayes methods in important statistical assessments. Policy-relevant statistical assessments involve synthesis of information from a set of related components such as medical clinics, geographic regions or research studies. Typical assessments include inference for individual parameters, synthesis over the collection of components (for example, the parameter histogram) and comparisons among parameters (for example, ranks). The relative importance of these goals depends on the context. Bayesian structuring provides a guide to valid inference. For example, while posterior means are the 'obvious' and optimal estimates for individual components under squared error loss, their empirical distribution function (EDF) is underdispersed and never valid for estimating the EDF of the true, underlying parameters. Effective histogram estimates result from optimizing a loss function based in a distance between the histogram and its estimate. Similarly, ranking observed data usually produces poor estimates and ranking posterior means can be inappropriate. Effective estimates should be based on a loss function that caters directly to ranks. Using examples of 'borrowing information', shrinkage and the variance/bias trade-off we motivate Bayes and empirical Bayes analysis. Then, we outline the formal approach and discuss 'triple-goal' estimates with values that when ranked produce optimal ranks, for which the EDF is an optimal estimate of the parameter EDF and such that the values themselves are effective estimates of co-ordinate-specific parameters. We use basic models and data analysis examples to highlight the conceptual and structural issues. PMID- 10474156 TI - Some adaptive monte carlo methods for Bayesian inference. AB - Monte Carlo methods, in particular Markov chain Monte Carlo methods, have become increasingly important as a tool for practical Bayesian inference in recent years. A wide range of algorithms is available, and choosing an algorithm that will work well on a specific problem is challenging. It is therefore important to explore the possibility of developing adaptive strategies that choose and adjust the algorithm to a particular context based on information obtained during sampling as well as information provided with the problem. This paper outlines some of the issues in developing adaptive methods and presents some preliminary results. PMID- 10474157 TI - On non-parametric maximum likelihood estimation of the bivariate survivor function. AB - The likelihood function for the bivariate survivor function F, under independent censorship, is maximized to obtain a non-parametric maximum likelihood estimator &Fcirc;. &Fcirc; may or may not be unique depending on the configuration of singly- and doubly-censored pairs. The likelihood function can be maximized by placing all mass on the grid formed by the uncensored failure times, or half lines beyond the failure time grid, or in the upper right quadrant beyond the grid. By accumulating the mass along lines (or regions) where the likelihood is flat, one obtains a partially maximized likelihood as a function of parameters that can be uniquely estimated. The score equations corresponding to these point mass parameters are derived, using a Lagrange multiplier technique to ensure unit total mass, and a modified Newton procedure is used to calculate the parameter estimates in some limited simulation studies. Some considerations for the further development of non-parametric bivariate survivor function estimators are briefly described. PMID- 10474159 TI - Brief communications. PMID- 10474158 TI - Assessment and comparison of prognostic classification schemes for survival data. AB - Prognostic classification schemes have often been used in medical applications, but rarely subjected to a rigorous examination of their adequacy. For survival data, the statistical methodology to assess such schemes consists mainly of a range of ad hoc approaches, and there is an alarming lack of commonly accepted standards in this field. We review these methods and develop measures of inaccuracy which may be calculated in a validation study in order to assess the usefulness of estimated patient-specific survival probabilities associated with a prognostic classification scheme. These measures are meaningful even when the estimated probabilities are misspecified, and asymptotically they are not affected by random censorship. In addition, they can be used to derive R(2)-type measures of explained residual variation. A breast cancer study will serve for illustration throughout the paper. PMID- 10474160 TI - Expression of Mmp-9 and related matrix metalloproteinase genes during axolotl limb regeneration. AB - One of the earliest events in limb regeneration is the extensive remodeling of the extracellular matrix (ECM). Matrix metalloproteinases (MMPs) are a family of matrix degrading enzymes that have been identified in both normal and disease states. Using RT-PCR and cDNA library screening, we have isolated sequences homologous to four different Mmp genes. The spatial and temporal expression of one of these, Mmp-9, has been analyzed during axolotl limb regeneration. Northern blot analysis identifies a 3.8 kb transcript that is abundantly expressed during regeneration, and whole-mount in situ hybridization has uncovered an unusual bi phasic expression pattern. The first phase begins at 2 hours after amputation, and expression is confined to the healed wound epithelium. This phase continues for 2 days, showing peak expression at 14 hours after amputation. This early phase may be needed to retard reformation of the basal lamina of the epidermis, and thereby facilitate the epidermal-mesenchymal interactions required for successful regeneration. The second phase begins a few days later when a small blastema has formed. During this phase, expression is in the mesenchyme, localized to cells around the tips of the cut skeletal elements. This expression is maintained through several stages until redifferentiation begins. The timing and position of the second phase of expression is consistent with a role for Mmp 9 in the removal of damaged cartilage matrix. We have also discovered that the time of onset of Mmp-9 expression is sensitive to denervation, which causes a delay of several hours. Finally, retinoids, known for their dramatic effects on the pattern of regenerating limbs, can cause a down regulation of Mmp-9 expression. Dev Dyn 1999;216:2-9. PMID- 10474161 TI - Dlx-5 in limb initiation in the chick embryo. AB - Dlx-5 is a vertebrate homolog of the Drosophila Distal-less gene, one of the first genetic signals for limb formation in the fly. In the present study we have explored the possible role of Dlx-5 in limb initiation in the chick embryo. At stage 14 which is well before the initial formation of limb buds Dlx-5 is highly and specifically expressed in the ectoderm of the presumptive wing and leg forming regions of the lateral plate, but not in the intervening non-limb forming prospective flank. Thus, Dlx-5 expression distinguishes the limb-forming territories prior to limb budding, and is one of the first molecular markers of vertebrate limb initiation. Furthermore, Dlx-5 expression is induced in the non limb-forming flank within 12 hours after implantation of an FGF2-soaked bead, a procedure that results in the induction of an ectopic limb. The rapid induction of Dlx-5 expression in response to a signal which ultimately leads to supernumerary limb formation is consistent with a role for Dlx-5 in limb initiation. We have also examined the expression of Dlx-5 in the limb buds of amelic limbless mutant chick embryos, which undergo normal limb formation but do not form an AER and thus fail to undergo further outgrowth. Dlx-5 is transiently expressed by the ectoderm of emergent limbless limb buds, consistent with a role for Dlx-5 in limb initiation. Together, our results suggest that Dlx-5 may be involved in the specification of the limb territories of the lateral plate, and in the initial formation of the limb bud from these regions. Dev Dyn 1999;216:10 15. PMID- 10474162 TI - Expression of the Mf1 gene in developing mouse hearts: implication in the development of human congenital heart defects. AB - The transcription factor FKHL7 gene has recently been associated with the anterior segment dysgenesis disorder of the eye known as Axenfeld-Rieger anomaly (ARA). A growing body of evidence indicates that mutations in FKHL7 cause not only defects in the anterior segment of the eye but defects in the heart valves and septa as well. In order to evaluate its contribution to normal heart septation and valve formation, expression of the mouse homologue Mf1 in embryonic hearts was analyzed by in situ hybridization. A weak but significant level of Mf1 expression could be detected in the endocardium of mouse embryos as early as day 8.5 post-conception (p.c.). Mf1 expression was undetectable in the hearts of day 9.5 p.c. embryos, but by day 10.5-11 p.c., Mf1 transcripts could be found again in the endocardium of both the atrium and ventricle and a relatively strong signal was observed in the dorsal portion of the septum primum, in what appeared to be the spinal vestibule. At day 13 p.c. when aortic and pulmonary trunks are separated, relatively more Mf1 transcripts were detected in the leaflets of aortic, pulmonary, and venous valves, the ventral portion of the septum primum, as well as in the single layer of cells on the edges of the atrioventricular cushion tissues. Surprisingly, there was no signal detected in the developing interventricular septum. At day 15 p.c., overall Mf1 signals were greatly decreased. However, significant levels of expression could still be observed in the atrial septum, the tricuspid valve, the mitral valve, and in the venous valve but not in the interventricular septum. The temporal and spatial expression patterns of the Mf1 gene in developing mouse hearts suggest that Mf1 may play a critical role in the formation of valves and septa with the exception of the interventricular septum. This is further supported by our studies showing that mutations in the FKHL7 gene were associated with defects in the anterior segment of the eye as well as atrial septal defects or mitral valve defects. Dev Dyn 1999;216:16-27. PMID- 10474163 TI - Stimulation of coronary vasculogenesis/angiogenesis by hypoxia in cultured embryonic hearts. AB - Hypoxia is known to stimulate vascular growth by up-regulating vascular endothelial growth factor (VEGF), but little is known about the function of hypoxia in the development of the coronary vasculature, and the relationship between hypoxia and VEGF in this event. To test the effects of hypoxia and VEGF on coronary vasculogenesis/angiogenesis in the developing heart, ventricles from 6-day-old quail embryos were cultured on three-dimensional collagen gels. After 2 days of growth in normal medium and 1 day of starvation in low serum medium (0.5% fetal bovine serum), the heart explants were further cultured under various oxygen levels for another 24, 48, and 72 hr. Angioblasts and endothelial cells, which migrated out from the heart explants, were identified by QH1 antibody using immunofluorescence and confocal microscopy. In the normoxic culture environment, the endothelial cells began to proliferate and migrate out from the heart explants after 3 days of growth; they formed tubes mainly after another 72 hr. In contrast, this vascular growth was accelerated under hypoxic conditions, as evidenced by increased tube formation with significant differences observed at 48 hr. On the other hand, hyperoxia delayed this process. Reverse transcription polymerase chain reaction results indicated that VEGF (including VEGF(122), VEGF(166), and VEGF(190)) was up-regulated in the heart explants under hypoxia and down-regulated under hyperoxia. VEGF neutralizing antibody added to the culture medium partially blocked this vascular growth. We conclude from this study that hypoxia can stimulate or up-regulate coronary vasculogenesis/angiogenesis and that VEGF signaling plays a major role in this event. Dev Dyn 1999;216:28-36. PMID- 10474164 TI - Ventrally emigrating neural tube cells contribute to the formation of Meckel's and quadrate cartilage. AB - A population of multipotential neuroepithelial cells originating in the ventral portion of the hindbrain neural tube has been shown recently to emigrate at the site of attachment of the trigeminal nerve. These ventrally emigrating neural tube cells populate the mesenchyme of the first pharyngeal (branchial) arch. Because the Meckel's and the quadrate cartilage develop from this mesenchyme, we sought to determine whether these ventrally emigrating neural tube cells contributed to their development. The ventral neural tube cells were tagged with a replication-deficient retroviral vector containing the LacZ gene. This method permanently labels the descendents of the neural tube cells; thus, they can be subsequently tracked during development. The viral concentrate was microinjected into the lumen of the rostral hindbrain of chick embryos, after the emigration of neural crest is finished, on embryonic day 2 (stage 14). In control embryos, the virus was placed on top of the neural tube. Embryos were killed on days 3, 4, and 7 and processed for the detection of LacZ-positive cells. By day 7, the Meckel's and the quadrate cartilage can be easily recognized. LacZ-positive cells were seen in both cartilages. They were located in perichondrium and in the cartilage. Immunostaining with the neural crest cell marker HNK-1 indicated that the LacZ positive cells were HNK-1 negative. The HNK-1-positive neural crest-derived cells were located in the cartilage but not in the perichondrium. These results indicate that the chondrocytes in the Meckel's and the quadrate cartilage differentiate from two sources of cells; the ventrally emigrating neural tube cells and the neural crest. The developmental significance of differentiation of cartilage from the ventral neural tube cells and of the heterogeneous origin of chondrocytes in morphogenesis remains to be established. Dev Dyn 1999;216:37-44. PMID- 10474165 TI - WNT11 promotes cardiac tissue formation of early mesoderm. AB - Cardiac tissue in the bird is derived from paired regions of lateral mesoderm within the anterior half of the embryo (Rawles [1943] Physiol. Zool. 16:22-42; Stalsberg and DeHaan [1969] Dev. Biol. 19:128-159). Previously, we reported that WNT11 is expressed in early avian mesoderm in a pattern that overlaps with the precardiac regions. To examine whether this molecule may play a role in promoting cardiogenesis, we cultured tissue explants from microdissected HH stage 4, 5, and 6 quail embryos. The isolated tissue consisted of both the mesoderm and endoderm layers from either anterior precardiac or posterior noncardiogenic regions of the embryo. As a necessary control for examining the ability of WNT11 to convert noncardiogenic mesoderm to cardiac tissue, we compared the cardiogenic potential of anterior and posterior regions. For stages 5 and 6, our results were consistent with what has been previously reported (Rawles [1943] Physiol. Zool. 16:22-42; Sugi and Lough [1994] Dev. Dyn. 200:155-162); as anterior mesoderm becomes contractile, while posterior mesoderm does not produce cardiac tissue. Surprisingly, when we examined stage 4 embryos both anterior and posterior regions gave rise to cardiac tissue in culture. To determine whether WNT11 could promote cardiac differentiation in tissue that was noncardiogenic, this molecule was ectopically expressed or added to mesoderm/endoderm explants obtained from stage 5 or 6 posterior tissue. Transfection of stage 5 posterior tissue with a WNT11 expression plasmid provoked the appearance of cardiomyocytes in 33% of the explants; half of which were contractile. Similarly transfected stage 6 posterior explants did not demonstrate cardiac differentiation. More dramatic results were obtained when noncardiogenic tissue was exposed to conditioned media containing soluble WNT11; as 63% and 33% of posterior stage 5- or stage 6-derived explants underwent cardiac differentiation. Together, these results indicate that WNT11 can promote cardiac development within noncardiac tissue. The expression of WNT11 in anterior mesoderm of early gastrula stage embryos suggests it may play a role in the formation of the vertebrate heart. Dev Dyn 1999;216:45-58. PMID- 10474166 TI - Expression of genes of type I and type II collagen in the formation and development of the blastema of regenerating newt limb. AB - We cloned cDNAs of alpha1(I) and alpha1(II) collagen, and studied their expression profiles in regenerating limbs of newts, Cynops pyrrhogaster. The expression of the alpha1(I) gene was markedly up-regulated at the early bud stage of the blastema. In situ hybridization experiments revealed that the alpha1(I) gene was expressed in not only mesenchymal cells of the blastema, but also the basal cells of the wound epidermis at the wound healing stage when the epidermal basement membrane was absent. This unique expression continued until 21 days (late bud stage), while the basement membrane began to form at 14 days. These results indicate biochemical differences between the wound and normal epidermis, and suggest the direct involvement of the former in the synthesis of blastemal matrices of type I collagen. Actually, immunohistochemistry revealed that type I collagen began to be deposited beneath the wound epidermis at 8 days, and accumulated there and around blastemal mesenchymal cells at 14 to 21 days. Undifferentiated mesenchymal cells associated with the amputated muscle fibers actively expressed the alpha1(I) gene. Mesenchymal cells in the central region of blastemas deposited type I collagen fibers around them. Concomitantly with the appearance of prechondrocytes, the alpha1(II) collagen gene became activated. The present study clearly shows that the expression of the genes of both type I and type II collagen in blastemal cells is temporally and regionally well-regulated in a cooperative manner. Dev Dyn 1999;216:59-71. PMID- 10474167 TI - Genomic organization and embryonic expression of the mouse fibroblast growth factor 9 gene. AB - Fibroblast growth factor 9 (FGF9), originally cloned as glial-activating factor from human glioma cells, is expressed in adult rat brain and kidney. Here we report the chromosomal localization, genomic organization, and embryonic expression pattern of the mouse Fgf9 gene. Fgf9 maps to chromosome 14 near the Ctla6 locus. The gene spans more than 34 kb and contains three exons and two introns. Translation initiation occurs in exon 1, and translation termination occurs in exon 3. Fgf9 RNA was detected during mouse embryogenesis in several tissues in which Fgf gene expression has not been previously described, including intermediate mesoderm of late-stage gastrulation, ventricular myocardium, lung pleura, skeletal myoblasts in the early limb bud, spinal cord motor neurons, olfactory bulb, and gut lumenal epithelium. Fgf9 is coexpressed with other Fgf genes in some skeletal myoblasts, in limb apical ectoderm, in craniofacial ectoderm, and in the retina, inner ear, and tooth bud. Dev Dyn 1999;216:72-88. PMID- 10474168 TI - Lexical decision in Parkinson's disease: A comment on Spicer, Brown, and Gorell (1994), and McDonald, Brown, and Gorell (1996) AB - Recent semantic priming investigations conducted by Spicer, Brown and Gorell (1994) and McDonald, Brown and Gorell (1996) reported hyperpriming in persons with Parkinson's disease (PD) and provided evidence supporting impaired set shifting as the underlying cause. This paper discusses notable priming behaviours exhibited by the normal control groups employed by Spicer and McDonald and colleagues. The argument is developed that these studies only measured attention dependent semantic processing and, therefore, their results do not speak to issues of automatic semantic activation in PD as originally proposed by Spicer et al. nor to putative deficits in set-shifting as proposed by McDonald et al. The implications for future priming research in PD are also discussed. PMID- 10474170 TI - A familial factor in the development of face recognition deficits. AB - The face is probably the single most important visual object. It is the main cue for identifying familiar people and crucial in nonverbal communication. Face recognition can be demonstrated in newborn babies, suggesting that face recognition is to some degree 'hard-wired' in the brain. Also, neurological disease can selectively disrupt the ability to recognize faces. Apart from acquired face recognition deficits, developmental cases have been reported. This study presents a family in which the father and two of his daughters are very poor at recognizing faces, demonstrating for the first time a familial factor in developmental face recognition deficit. PMID- 10474169 TI - Lexical priming on Neely's (1977) paradigm in Parkinson's disease: where do we stand? AB - In two published studies, we used a variant of Neely's (1977) lexical decision paradigm to study shifts of attention and automatic lexical activation in nondemented individuals with Parkinson's disease (McDonald, Brown, & Gorell, 1996; Spicer, Brown, & Gorell, 1994). Arnott and Chenery (in press) noticed differences between Neely's results and the results we observed in our control group that raise questions about some of the conclusions presented in the McDonald et al. (1996) and Spicer et al. (1994) papers. Even when considering the important differences between Neely's (1977) results and those in our control groups, we argue that our results support the conclusions of normal automatic semantic activation and deficient set-shifting in Parkinson's disease. We also introduce the notion of generalized priming to account for some of the priming effects observed in our studies. PMID- 10474171 TI - Dysfunction in visual object recognition of patients with unilateral brain damage under the element presentation condition. AB - The ability of integrating visual elements of 12 patients with right hemisphere damage (RHD), 14 patients with left hemisphere damage (LHD), and 20 normal young adults was investigated to specify the visual dysfunction processes of patients with unilateral brain damage. Both RHD and LHD groups demonstrated clear deficit in recognizing the familiar objects of which elements were cyclically presented at various time intervals. Moreover, in the nonsequentail element presentation condition the spatial and temporal integration of the RHD group was impaired relative to the LHD and normal groups. The visual degradations of patients with brain damage were discussed from the point of view of information processing stage as well as serial and parallel processing. PMID- 10474172 TI - Conceptual and perceptual contributions to the acquisition and retention of text specific reading skill in Alzheimer's disease. AB - In the first experiment, patients with Alzheimer's disease (AD) and normal control (NC) participants read each of two passages three times and then again after either a 1-min or 10-min delay interval. Reading time decreased with practice for both groups, and this facilitation was text-specific. Although both groups retained the facilitation in reading speed after 1 min, it was reduced only in the AD group after 10 min. In the second experiment, the same task was repeated with new passages that encouraged the use of either conceptual or perceptual processing during reading. Patients with AD acquired the facilitation in reading speed normally in both conditions, but were impaired in retaining it over a 10-min delay interval only in the conceptually-weighted condition PMID- 10474173 TI - A test of odor fluency in patients with Alzheimer's and Huntington's disease. AB - In the present study, an olfactory analog to the verbal fluency test was designed and administered to 40 patients with Alzheimer's disease (AD) and 11 patients with Huntington's disease (HD). Because onset of AD is typically in the sixties while onset of HD is typically in the mid-thirties, the patient groups had their own control group, an older (ONC) and younger (YNC) control group, respectively. Both control groups included 40 participants who were age- and education-matched to their respective patient group. Odor threshold, odor identification, and odor fluency measures were administered to each participant. Results of the study indicate that patients with dementia perform more poorly on all three measures of olfactory functioning. Our results suggest that tests of odor memory show differential performance in healthy and demented patients, leading to the suggestion that tests of olfactory functioning may be useful in detection and diagnosis of neurodegenerative disease. PMID- 10474174 TI - Reaction time indicators of attention deficits in closed head injury. AB - The nature of deficits in attention in closed head injury (CHI) was studied by three reaction time (RT) paradigms given to 20 patients who had a CHI 2 or more years previously and to 25 controls. We studied the effects of temporal uncertainty by varying the length and regularity of the preparatory interval, the effects of stimulus modality uncertainty on simple RT to tones and lights, and the effects of response selection in choice RT. The CHI group showed slower and more variable RT than controls under all conditions. In addition, a long preparatory interval on the preceding trial retarded RT more in the CHI group, and they showed greater effects of stimulus modality uncertainty. Both of these findings suggest a difficulty in shifting attention to unexpected stimuli. These greater effects on RT of variations of attention or preparation in CHI may account for their greater within-subject variability possibly due to frontal lobe damage. PMID- 10474175 TI - WAIS-R prediction equations in patients with traumatic brain injury. AB - Arithmetic algorithms for predicting premorbid Full Scale IQ were evaluated in a sample of 125 brain injured adults who had been evaluated within two months of their injury, and then again one year later. FSIQ at post-test was used as the criterion variable. The prediction equation, based on demographic information only (Barona, Chastain, & Reynolds, 1984), proved to be a modest predictor of premorbid FSIQ. The BEST-3 (Vanderploeg & Schinka, 1995), a prediction equation that incorporates demographic information and performance data, was sensitive to the severity of the brain injury. Subsequent analyses found the BEST-3 to be a better predictor of recovery of function than it was an estimate of premorbid functioning. This study demonstrates the need for flexibility in the interpretation of results, as what was thought to be a prediction equation for premorbid functioning was better viewed as an estimate of recovery. PMID- 10474176 TI - Modality specificity of memory span tasks among Zairian children: A developmental perspective. AB - Digit span (DS) and visual memory span (VMS) tasks have been shown to be distinct in young children but fail to show modality specificity in older American children and adults (Fastenau, Conant, & Lauer, 1997, 1998). The present study examines the possibility that processing of VMS becomes automatized with written language training. In a sample of 139 Zairian children, separate factor analyses on K-ABC data for younger children (6.1-8.5 years) and older children (8.6-12.8 years) yielded two-factor solutions in both cases, which appear to represent verbal and visual-spatial abilities. Modality specificity of the visual span task is supported in both age groups. Findings of modality specificity in both Zairian age groups provides support for the theoretical distinction between verbal and visual memory span. Continued modality specificity of visual memory span tasks in older Zairian children suggests that the increased verbal loadings of these tasks seen for older American children may reflect differences in written language development. PMID- 10474177 TI - Time perception following unilateral amobarbital injection in patients with temporal lobe epilepsy. AB - Estimates of elapsed time were obtained from 53 patients with unilateral temporal lobe epilepsy (Left TLE = 27; Right TLE = 26) following Wada (intracarotid amobarbital) assessment. After resolution of drug effects, patients were asked to estimate how much time had passed since amobarbital administration. Estimates were also obtained from 24 healthy control subjects using the same cognitive tasks over a similar time frame. Elapsed time was significantly underestimated by both left and right TLE groups following right hemisphere injection. In addition, there was an interaction effect involving patient group, side of injection, and sequence of injection. Left TLE patients, consistent with normal controls, made more accurate time estimates when they could anticipate the estimation task following the second amobarbital administration. More accurate time estimates, however, occurred only when left hemisphere injection was second in sequence. In contrast, right TLE patients did not improve regardless of the order of injection. These results suggest that right hemisphere function plays a critical role in the accuracy of time estimations of intermediate temporal duration and that interhemispheric interaction may be required to make accurate retrospective temporal judgments. These findings are discussed in the context of the growing evidence for a right-hemispheric attentional network. PMID- 10474178 TI - Is there a syndrome of tuberothalamic artery infarction? A case report and critical review. AB - Short-term post-acute neuropsychological, neurological, and neuroradiological test results and a 16-month follow-up of a 65-year-old patient with a right hemisphere ischemic lesion in the tuberothalamic area of vascular supply are reported. During a 6-week period of examinations the originally left- but trained right-handed patient exhibited fluctuating neuropsychological disorders including aphasia, visuo-perceptive and visuoconstructive disorders, and memory and attention deficits. In the follow-up examination the patient exhibited no aphasia and significant improvements in most neuropsychological tasks. Based on three dimensional reconstruction of MRI, lesion topography and involvement of thalamic nuclei were established. We discuss the neuropsychological and neurological symptoms of the present case against the background of the 'syndrome of unilateral tuberothalamic artery territory infarction' proposed by Bogousslavsky and coworkers (1986) and the neuropsychological literature on unilateral ischemic anterior/anterolateral thalamic infarction. PMID- 10474179 TI - Revisiting indirect bonding. PMID- 10474180 TI - Comparison of dynamic and integrated light-scattering techniques in the study of the interaction of Candida rugosa lipase with DPPC liposomes. AB - Dynamic light-scattering (DLS) and wide angle integrated light-scattering (WAILS) spectroscopies were evaluated in the study of binding of Candida rugosa lipase (CRL) with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) liposomes. The use of cumulants analysis on DLS data allowed for the determination of general lipase liposome-binding trends. Particle intensity distributions obtained from DLS data by a discrete inversion method revealed the different populations created upon lipase-liposome interactions. Using a discrete inversion technique on WAILS data, not only these populations could be differentiated but also accurate number distributions were obtained in short periods of time. Both DLS and WAILS are excellent tools for the study of lipase binding to lipid vesicles; however, care must be exercised in the analysis of the experimental data whenever particle size distributions are multimodal. The selection of the light scattering technique will depend on the information required. PMID- 10474181 TI - Low doses of alpha- and gamma-radiation enhance DNA thermal stability. AB - Isolated calf thymus DNA in buffered solutions has been exposed to 0-150 Gy of alpha- and gamma-radiation. The effects of alpha- and gamma-radiation on the thermal stability and electrophoretic mobility of the DNA molecules have been studied by UV spectroscopic 'melting' and Pulsed Field Gel Electrophoresis (PFGE), respectively. The observed thermal denaturation parameters were fitted to the energy propagation descriptive model. The experimental results for the samples exposed to relatively low (low) doses indicate an increased thermal stability and a reduced mobility over that of the controls. The expected overall degradation of the DNA molecules was confirmed for the samples exposed to high doses. Our results are in good agreement with the predictions of the energy propagation model, which now is also tested in the low dose region and for an additional type of ionising radiation (alpha-particles). Our findings are consistent with conformational changes at low doses resulting in a DNA form characterised by localised alterations, which affect the energy flow along the DNA molecule. PMID- 10474182 TI - Contractile and protractile coordination within an actin filament sliding on myosin molecules. AB - An actin filament exhibits distortions longitudinally when it slides upon myosin molecules. We observed that the actin filament demonstrated contractile distortions at low ATP concentrations and protractile distortions at high concentrations. Temporal development of such distortions was identified, by tracing each of several speckled fluorescent markers attached to the actin filament. Close association of the sliding movement to the moving distortions of an actin filament suggests the presence of a unitary mechanism regulating the apparently two different modes of dynamic movement. PMID- 10474183 TI - The Tol proteins of Escherichia coli and their involvement in the uptake of biomolecules and outer membrane stability. AB - The Tol proteins of Escherichia coli are involved in outer membrane stability. They are also required for the uptake of the group A colicins and the translocation of filamentous phage DNA into the cytoplasm. The tol-pal genes constitute two operons in the E. coli genome, orfltolQRA and tolBpalorf2. The TolQ TolR TolA proteins form a complex in the cytoplasmic membrane, while TolB and Pal interact near the outer membrane. Most of the amino acid residues of TolA, TolB, TolR and Pal are localized in the periplasm. Recent advances in the knowledge of interactions of Tol-Pal proteins with other envelope components, or with group A colicins, are presented, together with current hypotheses about the role of the Tol proteins in outer membrane stability. PMID- 10474184 TI - Effect of Cu,Zn superoxide dismutase disruption mutation on replicative senescence in Saccharomyces cerevisiae. AB - The role of oxidative damage in determining the replicative lifespan of Saccharomyces cerevisiae was investigated using a wild-type haploid laboratory yeast and a Cu,Zn superoxide dismutase (sod1) mutant derivative on glucose, ethanol, glycerol and galactose media. SOD1 expression was necessary to ensure longevity on all carbon sources tested. Whilst carbon source and SOD1 gene expression do influence yeast lifespan, the relationship between the two factors is complex. PMID- 10474185 TI - A Candida albicans metallopeptidase degrades constitutive proteins of extracellular matrix. AB - Among potential virulence factors of Candida albicans, enzymes seem to play an important role. Many studies concern the secreted aspartic proteinases (saps), and the degradation of some components of the subendothelial extracellular matrix by the isoenzyme sap2 has been proved. Nevertheless, other proteolytic enzymes could be involved in the pathogenicity of the yeast. We studied the degradation of four constitutive proteins of the extracellular matrix: type I and IV collagens, laminin and fibronectin, by a 95-kDa metallopeptidase, localised in the cell wall of C. albicans. Each of these constituents was incubated with the purified enzyme and its degradation products analysed by an electrophoretic method. We observed that type I collagen and fibronectin were totally degraded by the enzyme whereas type IV collagen and laminin were only partially degraded. The C. albicans metallopeptidase may play a role in the degradation of the subendothelial extracellular matrix components. This enzyme could facilitate the migration of the yeast in the tissues after crossing the endothelial layer, allowing the fungal invasion of target organs. PMID- 10474186 TI - Enolase is present in the cell wall of Saccharomyces cerevisiae. AB - Non-covalently attached or soluble cell wall proteins of Saccharomyces cerevisiae were extracted using a high pH/2-mercaptoethanol procedure and were separated for peptide sequencing using 2D-PAGE. A partial N-terminal sequence of a major protein spot was obtained and showed high identity with enolase gene products. Western blotting with an anti-enolase antibody confirmed that enolase was present in the cell wall extract. Biotinylation of cells prior to protein extraction with a membrane impermeable biotinylating agent confirmed that the detection was not owing to cell lysis during extraction. Transmission immunoelectron microscopy showed enolase to be present in the cell wall. Enolase contains no known secretion signal that would localize it to the cell wall. Thus S. cerevisiae must have further mechanisms for targeting proteins to the cell wall. PMID- 10474187 TI - Identification of nodulation promoter (nod-box) regions of Rhizobium galegae. AB - A hybridisation analysis of a genomic clone library of Rhizobium galegae HAMBI 1174 located four EcoRI fragments homologous to the nod-box promoter sequence of Sinorhizobium meliloti in two separate gene regions. Two of the five nod-boxes detected in the R. galegae genome were carried on a single cosmid clone, pRg30, upstream from the nodABCIJ and nodF genes, whereas the other three nod-boxes were carried on a different cosmid clone, pRg10. Hybridisations with various nod gene probes from S. meliloti and Rhizobium leguminosarum species detected a nodD homolog in pRg10. The sequence data obtained from regions adjacent to each nod box in pRg10 confirmed the presence of a second nodD in the R. galegae genome and, in addition, revealed the presence of nodN, nodU, dctA nifH and nifQ-like genes in pRg10. Thus, by using a promoter-specific nod-box probe we could identify a new region carrying genes involved in nitrogen fixation and host specificity functions. PMID- 10474188 TI - The sequence of the non-haemolytic enterotoxin operon from Bacillus cereus. AB - The non-haemolytic enterotoxin from Bacillus cereus has been sequenced. It is composed of three components, non-haemolytic enterotoxin A, B and C of 41.0, 39.8 and 36.5 kDa, respectively. Transcription of the operon seems to be positively regulated by plcR, a gene that also regulates phospholipase C expression. There is substantial similarity between the three proteins of non-haemolytic enterotoxin and between the non-haemolytic enterotoxin and haemolytic enterotoxin proteins. PMID- 10474189 TI - Diaminodiphenylsulfone resistance of Mycobacterium leprae due to mutations in the dihydropteroate synthase gene. AB - The nucleotide sequence analysis of the dihydropteroate synthase (DHPS) gene of six diaminodiphenylsulfone-resistant Mycobacterium leprae strains revealed that the mutation was limited at highly conserved amino acid residues 53 or 55. Though the mutation at amino acid residue 55 or its homologous site has been reported in other bacteria, the mutation at residue 53 is the first case in bacteria. This is the first paper which links the mutations in DHPS and sulfonamide resistance in M. leprae. This finding is medically and socially relevant, since leprosy is still a big problem in certain regions. PMID- 10474190 TI - Disruption of the mycobacterial cell entry gene of Mycobacterium bovis BCG results in a mutant that exhibits a reduced invasiveness for epithelial cells. AB - Mycobacteria belonging to the Mycobacterium tuberculosis complex have the ability to invade and replicate in non-phagocytic cells, an event that requires the presence of bacterial surface components capable of triggering a cell response and the subsequent internalization of the microorganism. In this study, we report the sequencing of the mycobacterial cell entry gene (mce) of Mycobacterium bovis bacillus Calmette-Guerin (BCG) and the generation and characterization of a mutant BCG strain with an inactivated mce gene, by homologous recombination with double cross-over. This mutant strain does not express the mycobacterial cell entry protein (Mce) and exhibits a reduced ability to invade the non-phagocytic epithelial cell line HeLa as compared to wild-type BCG. PMID- 10474191 TI - Secondary structure of Streptococcus downei GTF-1 glucansucrase. AB - Multiple sequence alignment and structure prediction of glucansucrases produced by oral streptococci and Leuconostoc mesenteroides showed that all have common structural features, with three major domains. There is no conservation of primary sequence or structure in the N-terminal variable region. Sequence-based structure prediction combined with circular dichroism spectrum analysis of purified truncated forms of Streptococcus downei GTF-I revealed that the core catalytic region has a defined structure consistent with the proposed (alpha/beta)8-barrel structure. The C-terminal domain is a mixed structure with significant amounts of beta-sheet and random-coil. This information contributes to the development of our understanding of structure-function relationships in glucansucrases. PMID- 10474192 TI - Detection of genes for periplasmic nitrate reductase in nitrate respiring bacteria and in community DNA. AB - A nested PCR primed by four degenerate oligonucleotides was developed for the specific amplification of sequences from the napA gene encoding the periplasmic nitrate reductase. This approach was used to amplify fragments of the napA gene from 10 Pseudomonas species and one Moraxella sp., previously shown to be able to express the periplasmic nitrate reductase activity, from Rhodobacter capsulatus and from community DNA extracted from a fresh-water sediment. Amino acid sequences encoded by the napA fragments were compared to one another and to the corresponding regions of related enzymes. This comparison indicates that the amplification protocol is specific for its intended target. The napA sequences amplified from community DNA were tightly clustered, which may indicate a degree of homogeneity in the sediment community. All tested Gram-negative strains capable of aerobic nitrate respiration were found to have periplasmic nitrate reductase genes. However, some strains which have and express the genes are incapable of aerobic nitrate respiration. The PCR primers and amplification protocols described will be useful in future studies of nitrate respiring populations. PMID- 10474193 TI - Identification of genetic factors altering the SOS induction of DNA damage inducible yebG gene in Escherichia coli. AB - The yebG gene of Escherichia coli is a novel SOS regulon gene, but details of its regulation mechanism and biological function are not yet known. To characterize the regulation of yebG gene as a SOS gene, we identified the genetic factors affecting the SOS induction of yebG gene using yebG-lacZ operon fusion plasmid. We found that the SOS induction of yebG occurs as the cells enter into the stationary growth phase, but its induction is not observed in LB medium in the presence of 1% glucose. A stationary phase SOS induction of the yebG gene does not require the global regulator of stationary phase-specific genes, rpoS, or gyrA functions, but requires cya encoding the adenylate cyclase and hns encoding the histone-like protein H-NS functions. Our results demonstrated that the induction of a DNA damage-inducible yebG gene of E. coli is dependent on cyclic AMP and H-NS. PMID- 10474194 TI - Analysis of a ptsH homologue from Streptomyces coelicolor A3(2). AB - A ptsH homologue of Streptomyces coelicolor A3(2) was identified in the emerging genome sequence, cloned in Escherichia coli and the S. coelicolor HPr over produced and purified. The protein was phosphorylated in vitro in a phosphoenolpyruvate (PEP)-dependent manner by purified enzyme I (EI) from Bacillus subtilis, and much less efficiently in an ATP-dependent manner by purified HPr kinase, also from B. subtilis. There was no indication of ATP dependent phosphorylation of the purified protein by cell extracts of either S. coelicolor or Streptomyces lividans. Deletion of the ptsH homologue from the S. coelicolor and S. lividans chromosomes had no effect on growth when fructose was supplied as sole carbon source, and in S. coelicolor it had no effect on glucose repression of agarase and galactokinase synthesis, suggesting that the HPr encoded by this gene does not play an essential role in fructose transport nor a general role in carbon catabolite repression. PMID- 10474195 TI - Evidence for frequent OspC gene transfer between Borrelia valaisiana sp. nov. and other Lyme disease spirochetes. AB - Molecular polymorphism of the ospC gene has been reported in Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii, the spirochetes causing human Lyme borreliosis. To assess the genetic relationship between ospC genes from the recently described Borrelia valaisiana sp. nov. and other B. burgdorferi sensu lato species, the ospC genes from eight B. valaisiana isolates were amplified by PCR, cloned and sequenced. The ospC genes of three B. valaisiana isolates were identical, but clearly distinct from ospC genes from other Borrelia species. Four B. valaisiana isolates possessed ospC genes more related to those of B. garinii, and fell into a cluster representing B. garinii species in the phylogenetic tree. One isolate had an ospC gene encoding a protein identical to that of B. afzelii strain. Since five of the eight (62.5%) B. valaisiana isolates contained a gene highly homologous or even identical to ospC genes found among B. garinii and B. afzelii strains, our findings indicate that ospC gene transfer occurs between B. valaisiana and other Lyme disease spirochetes. PMID- 10474197 TI - Glycosylation of the Enterobacter cloacae outer membrane protein OmpX in eukaryotic cells. AB - The topological model of the Enterobacter cloacae outer membrane protein OmpX showed three putative glycosylation sites. When OmpX was expressed in bacteria that were cultured under aerated conditions, no glycosylation was observed. The coupling of carbohydrate chains to the ompX gene product was also investigated in the eukaryotic baculovirus expression system. For this purpose, a recombinant ompX gene-containing baculovirus was made. Infection of insect cells with this recombinant virus resulted in the production of sufficient amounts of OmpX to study glycosylation. In this system, all potential N-glycosylation sites of OmpX were utilized. Furthermore, it became clear that glycosylated OmpX was retained in the insect cells and was not secreted in the medium. Given the fact that OmpX plays a role in the invasion of E. cloacae in rabbit enterocytes, glycosylation of this protein occurring only under specific conditions may be involved in this process. PMID- 10474196 TI - Molecular cloning and analysis of a pleiotropic regulatory gene locus from the nystatin producer Streptomyces noursei ATCC11455. AB - A regulatory gene locus from Streptomyces noursei ATCC14455, the producer of the antifungal antibiotic nystatin, was cloned in Streptomyces lividans based on its ability to activate actinorhodin (Act) production in this host. Deletion and DNA sequencing analyses showed that a small gene, designated ssmA, located downstream of an afsR homologue (a known pleiotropic regulator) was responsible for the Act overproduction in S. lividans. Database searches for the ssmA gene product revealed its limited similarity to the AfsR2 regulatory protein from S. lividans and CREA catabolite repressor from Aspergillus nidulans. To study the effect of ssmA on nystatin production, this gene was either deleted from S. noursei genome, or placed under control of PermE* promoter and introduced in S. noursei. The properties of the corresponding strains indicate that ssmA is involved in regulation of growth and antibiotic production only in the media with certain carbon sources. PMID- 10474198 TI - Adherence and internalization of Streptococcus uberis to bovine mammary epithelial cells are mediated by host cell proteoglycans. AB - The role of cell glycosaminoglycans (GAG) on adherence and internalization of Streptococcus uberis to bovine mammary epithelial cells was evaluated by adherence/internalization competition assays, by removal of GAG from the host cell surface and by inhibition of GAG glycosylation in the host cell. Heparin (HEP), heparan sulfate (HSA), chondroitin sulfate A (CSA) and chondroitin sulfate B (CSB) inhibited adherence and internalization of S. uberis in a dose-dependent manner. However, inhibition was lower with CSA and CSB than that observed with HEP and HSA. Adherence and internalization were also inhibited upon treatment of mammary epithelial cells with GAG lyases. The greatest inhibition was observed with heparinase I. Tunicamycin, an inhibitor of mammalian cell glycosylation of cell surface glycoproteins, markedly inhibited internalization of S. uberis into mammary epithelial cells. Differences between strains were observed. These results suggest that a HSA proteoglycan receptor on the host cell surface may mediate S. uberis adherence to and internalization of bovine mammary epithelial cells. PMID- 10474199 TI - Comparative analysis of the LPS biosynthetic loci of the genetic subtypes of serovar Hardjo: Leptospira interrogans subtype Hardjoprajitno and Leptospira borgpetersenii subtype Hardjobovis. AB - Although Leptospira borgpetersenii subtype Hardjobovis and L. interrogans subtype Hardjoprajitno belong to different species, they are serologically indistinguishable and are therefore classified as serovar Hardjo. Since LPS is the major antigen involved in serological classification, this implies that the LPS of these subtypes is identical. Comparison of the LPS biosynthetic loci (rfb) of the subtypes revealed remarkable similarity, with 32 and 31 origins of replication (orfs) in the Hardjoprajitno and Hardjobovis rfb loci, respectively. The order and orientation of these orfs were identical with the exception of an additional orf in Hardjoprajitno between orfs 4 and 5 and intergenic sequences differing between the subtypes. The Hardjoprajitno rfb locus has been divided into four intercalated regions based on sequence similarity to other leptospiral rfb loci. orfJ1-orfJ14 as well as orfJ21-orfJ22 are more similar to regions of the rfb locus of L. borgpetersenii subtype Hardjobovis. orfJ15-orfJ20 as well as orfJ23-orfJ31 are almost identical to the corresponding orfs in L. interrogans serovar Copenhageni. We propose that the progenitor Hardjoprajitno strain, containing an rfb locus which closely resembled the Copenhageni locus, acquired orfs 1-14 and orfs 21-22 from subtype Hardjobovis resulting in two serologically indistinguishable subtypes of serovar Hardjo which in turn constituted the main bovine-adapted leptospiral serovar. PMID- 10474200 TI - Heat-inactivation of plasmid-encoded CI857 repressor induces gene expression from Ind- lambda prophage in recombinant Escherichia coli. AB - We have observed significant cell lysis upon temperature up-shift of recombinant Escherichia coli cultures harboring CI857-repressed lambda-based expression vectors. This event, that becomes evident about 30-40 min after the heat shock, takes place when using the lambda promoter system in Ind- lysogenic strains, but not in others commonly employed for recombinant gene expression. These results strongly suggest that the thermosensitive CI857 repressor, encoded by the expression vector, competes with CI Ind- molecules for binding to the prophage operator region, allowing for expression of lytic genes from the integrated Ind- viral genome upon temperature up-shift. Transcription of viral lytic genes does not include unspecific expression of a reporter sulA::lacZ gene fusion carried in the prophage genome. These results prompt, however, to carefully evaluate the limitations of expression systems based on pL/pR-CI857 in bacterial strains modified through lambda Ind- gene transfer vehicles. PMID- 10474201 TI - Urinary excretion of essential metals following intravenous calcium disodium edetate: an estimate of free zinc and zinc status in man. AB - Ethylenediaminetetraacetic acid (EDTA) is a powerful metal chelating agent used in the treatment of lead poisoning. EDTA also binds strongly to other metals. Thus, following intravenous infusion of CaNa2EDTA in healthy subjects the urinary excretion of calcium, copper, iron, magnesium and zinc were assessed. CaNa2EDTA significantly increased the urinary excretion of all metals except magnesium with greatest increases for iron (x 3.8 above baseline) and zinc (x 22). In addition, an in vitro dialysis study with a simplified serum showed that zinc (4.1 X 10(-3) mumol/h) was taken up more rapidly than iron (2.9 X 10(-3) mumol/h) by EDTA. The degree of binding of iron and zinc by EDTA depends on two factors: namely, the affinity of EDTA for Zn2+ and Fe3+, and the levels of unbound hydrated Zn2+ and Fe3+ ('free' ions). Despite differences in the rate of chelation of Zn2+ and Fe3+ by EDTA we show that the measurements of (a) circulating free iron, from routine clinical measurements of transferrin bound iron, and (b) the ratio of zinc:iron excreted in urine could provide an estimate of circulating free zinc, and thereby of zinc status, in man. In addition, EDTA treatment should be evaluated for patients with iron overload. PMID- 10474202 TI - Dietary minerals in the gastrointestinal tract: hydroxypolymerisation of aluminium is regulated by luminal mucins. AB - The regulation of mineral absorption in the gastrointestinal tract is poorly understood. Recent work has identified an intracellular metal-ion transporter but considerable evidence suggests that both soluble and mucosally associated luminal metal-binding ligands regulate initial uptake. Molecules ranging from low molecular weight organic acids to large glycoproteins have been suggested but a definite role for any such species has remained elusive. Here, a series of analytical techniques, allowing for this wide variation in potential binding ligands, was applied to the study of intestinal contents and tissue of rats following different feeding protocols. Aluminium, that has a low endogenous background and maintains a high concentration in the gastrointestinal tract, was investigated as a suitable dietary metal with hydrolytic behaviour similar, for example, to copper, iron and zinc. High resolution nuclear magnetic resonance spectroscopy identified a number of endogenous low molecular weight weak ligands that are secreted into the intestinal lumen. These may slow the rate of hydroxy polymerisation of hydrolytic metals, allowing their effective donation to less mobile, higher molecular weight binding ligands. Histochemical staining suggested that such species may be soluble mucins as these were consistently associated with luminal aluminium. Significantly, this interaction prevented hydroxy/phosphate precipitation of aluminium, even at supraphysiological levels of the element. This was confirmed with X-ray micro-analysis investigations of ex vivo luminal contents. Nevertheless, from phase distribution experiments, the majority (60-95%) of luminal aluminium was associated with the intestinal solid phase and further histochemistry confirmed this to be gelatinous mucus, chiefly as the mucosally adherent layer. All results suggest a major role for mucus in regulating the gastrointestinal absorption of aluminium. It is proposed that, initially, soluble luminal mucus prevents the hydroxy-precipitation of hydrolytic metals at intestinal pH, allowing their effective donation to the mucus layer. Based on the differing reported metal-mucus interactions, elements that bind well to mucus (Al3+, Fe3+), with kinetically slow rates of ligand exchange (Al3+ < Fe3+) will be less well absorbed than poorly bound elements with kinetically faster rates of ligand exchange (Cu2+, Zn2+ etc.). This mechanism would readily explain many of the reported observations on mineral availability, including the marked variation in absorption of different elements, the differential effects of dietary ligands on mineral uptake and the competition for absorption between different metals. PMID- 10474203 TI - Chloride and ethyl ester morpholine thiourea derivatives and their Ni(II) complexes. Crystal and molecular structures of the thiourea derivative L-leucine methyl ester and its complexes with Cu(II) and Pt(II). Growth of the pathogenic fungus Botrytis cinerea. AB - We have synthesized a series of ligands (1, 3, 4, 6 and 7) and some of their complexes with Ni(II), Cu(II) and Pt(II) (2, 5, 8 and 9). These compounds were studied and characterized by elemental analysis, IR and UV-Vis spectra, conductivity measurements in solution, FAB+/MS, 1H and 13C NMR, ESR, etc. Compound 7 crystallized in the orthorhombic space group P2(1)2(1)2(1), with Z = 4. Unit cell parameters were as follows: a = 21.307(2) A, alpha = 90 degrees, b = 12.498(1) A, beta = 90 degrees, c = 7.7232(4) A, gamma = 90 degrees. For seven of these compounds, the antifungal activity of a major pathogen responsible for important crop damage was studied. In general, inhibition by the ligands was higher than that of the complexes. When the thiourea was linked to some diethyl groups, the compounds showed higher antifungal activity than the morpholine groups. Compound 3 achieved total inhibition (100%). PMID- 10474204 TI - Comparative mechanism and toxicity of tetra- and dithiomolybdates in the removal of copper. AB - Tetrathiomolybdate (TTM) can be used as a specific chelator to remove copper (Cu) accumulating in the form bound to metallothionein (MT) in the livers of Wilson disease patients and Long-Evans rats with a cinnamon-like coat color (LEC rats). However, an adverse effect, hepatotoxicity, was observed occasionally on its clinical application. The mechanism underlying the adverse effect of TTM has been studied in comparison with dithiomolybdate (DTM), and a safer and more effective therapy by TTM was proposed based on the mechanism. The activity of glutamic pyruvic transaminase (GPT) in serum was shown to increase significantly on the treatment of Wistar rats with sulfide produced through hydrolytic degradation of TTM and DTM, the latter being more easily degraded. The hydrolytic degradation of TTM was enhanced under acidic conditions. Cu in Cu-containing enzymes such as Cu,Zn-superoxide dismutase (SOD) in liver and ceruloplasmin (Cp) in plasma was decreased by excessive thiomolybdates, the Cu being found in the plasma in the form of a Cu/thiomolybdate/albumin complex. The decreased amounts of Cu in SOD and Cp were explained by the sequestration of Cu from their chaperones by thiomolybdates rather than the direct removal of Cu from the enzymes. Although both TTM and DTM remove Cu from MT, DTM is not appropriate as a therapeutic agent for Wilson disease due to its easy hydrolysis and production of sulfide. PMID- 10474205 TI - Structural and cytotoxic study of new Pt(II) and Pd(II) complexes with the bi heterocyclic ligand mepirizole. AB - Pd(II) and Pt(II) complexes of formulae [MLCl2], where L = mepirizole, were synthesized and characterized. Two complexes were obtained and studied by different techniques: IR, 1H and 13C NMR and XPS spectroscopies and mass spectrometry (electrospray). The crystal structure of the complex cis-dichloro-4 methoxy-2-(5-methoxy-3-methyl- pyrazol-1-yl)-6-methyl-pyrimidinepalladium(II), [Pd(mep)Cl2], was studied by crystal X-ray diffraction. It consists of discrete molecules with planar geometry. Pd(II) ions are four-coordinated by two mepirizole nitrogen atoms (N1 from the pyrazole ring and N4 from the pyrimidine ring) and two chlorine atoms. The geometry of the PdN2Cl2 chromophore is a distortion of the square-planar coordination. Data from powder pattern X-ray diffraction of cis-dichloro-4-methoxy-2-(5-methoxy-3-methyl-pyrazol-1-yl)-6 methyl- pyrimidineplatinum(II), [Pt(mep)Cl2], demonstrated that the two complexes are isostructural. The cytotoxic activity of both Pd and Pt complexes was checked for six different tumor cell lines and was lower than that of cisplatin. The Pt bound to DNA was also checked and only a low percentage is able to cross the cell membrane. PMID- 10474206 TI - The interaction between human and bovine serum albumin and zinc studied by a competitive spectrophotometry. AB - The binding of zinc to human serum albumin (HSA) and bovine serum albumin (BSA) has been studied by a competitive spectrophotometry. The indictor ligand, 5-Br PAPS, as well as its metal complexes, ML and ML2, having intense colors with sufficient separate absorption peaks permits the analysis of the interaction between zinc and HSA or BSA. The results show that zinc binds to both albumins in the molar ratio of 1:1 at pH 6-8. The apparent association constants of Zn-HSA and Zn-BSA decrease with decreasing pH. The pH dependence of the constants for both Zn-HSA and Zn-BSA indicates the 1:1 competition between metal ions and hydrogen ions for the binding site. The intrinsic association constants of the metal and the proton are found to be log KZn-HSA = 7.1 +/- 0.2 and log KH-HSA = 8.2 +/- 0.3 for HSA and log KZn-BSA = 7.6 +/- 0.2 and log KH-BSA = 8.2 +/- 0.3 for BSA, respectively. PMID- 10474207 TI - DNA damage by copper(II) complexes: coordination-structural dependence of reactivities. AB - A variety of copper complexes with different structural features have been shown to bind double-helical DNA with binding constants of 10(4)-10(7) M-1 and to promote double-strand DNA damage upon reductant/H2O2 activation. The interaction of the Cu complex with DNA results in hyperchromism and shifts to longer wavelengths of the strongest transitions in the Cu complexes, as well as striking hypochromism or hyperchromism of DNA absorption at 260 nm. In the presence of DMPO as the spin trap, the solution of each copper complex exhibits typical four line ESR spectra of the hydroxyl radical by adding 2-mercaptoethanol and H2O2 to the solution. Quantitation by 2-deoxy-D-ribose shows that the competence of hydroxyl radical generation by the copper complexes upon reductant and H2O2 activation decreases in order, that is, Cu(HTCD)(2+)-Cu(Im)4Cl2-Cu(IDB) (NO3)2 > Cu(IDB)Cl2 > Cu(IDBt)Cl2. The copper complex-mediated hydroxyl radical, a powerful oxidant that attacks the adjacent DNA, is responsible for the DNA oxidative damage. The lambda DNA damage chemistry illustrates that the competence and selectivity of double-strand lambda DNA damage by the copper complexes are dependent on their geometric structures and types of ligands. The decreasing order of the DNA damage capacity by the present complexes is Cu(Im)4Cl2-Cu(IDB) (NO3)2 > Cu(HTCD)2+ > Cu(IDBt)Cl2 > Cu(IDB)Cl2. PMID- 10474208 TI - Lake Wobegon be gone! The "below-average effect" and the egocentric nature of comparative ability judgments. AB - Like the inhabitants of Garrison Keillor's (1985) fictional community of Lake Wobegon, most people appear to believe that their skills and abilities are above average. A series of studies illustrates one of the reasons why: when people compare themselves with their peers, they focus egocentrically on their own skills and insufficiently take into account the skills of the comparison group. This tendency engenders the oft-documented above-average effect in domains in which absolute skills tend to be high but produces a reliable below-average effect in domains in which absolute skills tend to be low (Studies 1 and 2). In Study 3, cognitive load exacerbated these biases, suggesting that people "anchor" on their assessment of their own abilities and insufficiently "adjust" to take into account the skills of the comparison group. These results suggest that the tendency to see oneself as above average may not be as ubiquitous as once thought. PMID- 10474209 TI - Spontaneous trait transference to familiar communicators: is a little knowledge a dangerous thing? AB - In most social cognition research participants are presented with unattributed information about unfamiliar stimulus persons. However, in the real world it is more common for people to learn about others through social communication and to know something about those with whom they communicate. Such issues are explored in relation to spontaneous trait transference, a phenomenon in which communicators are perceived as having traits that they merely describe in others. Three studies show that even familiar communicators became associated with, and attributed, the traits implied by their remarks. Surprisingly, these effects occurred even when the implied traits were incongruent with participants' prior knowledge about these communicators. The results are discussed in terms of (a) the generalizability of social cognition research, (b) the automaticity of simple associative phenomena, and (c) the interplay of simple associative and higher level processes. PMID- 10474210 TI - Close partner as sculptor of the ideal self: behavioral affirmation and the Michelangelo phenomenon. AB - This work incorporates concepts from the behavioral confirmation tradition, self tradition, and interdependence tradition to identify an interpersonal process termed the Michelangelo phenomenon. The Michelangelo phenomenon describes the means by which the self is shaped by a close partner's perceptions and behavior. Specifically, self movement toward the ideal self is described as a product of partner affirmation, or the degree to which a partner's perceptions of the self and behavior toward the self are congruent with the self's ideal. The results of 4 studies revealed strong associations between perceived partner affirmation and self movement toward the ideal self, using a variety of participant populations and measurement methods. In addition, perceived partner affirmation--particularly perceived partner behavioral affirmation--was strongly associated with quality of couple functioning and stability in ongoing relationships. PMID- 10474211 TI - Does trait coping exist? A momentary assessment approach to the evaluation of traits. AB - This study investigated the extent to which momentary reports of coping are a traitlike phenomenon and the validity of a questionnaire assessment of coping style. Participants (N = 96) completed the questionnaire, then monitored stressful events and coping activities for 2 days using a hand-held computer that administered a brief assessment every 40 min. Momentary reports exhibited a sizable traitlike component: Individual differences accounted for 42% of the variance for 2 coping items and 15-30% of the variance for 15 items. The questionnaire assessment of coping style was a poor predictor of average momentary coping; the coping style measure and the aggregated momentary measure of trait coping shared 23% of their variance for religion and 0-12% for the 16 other coping measures. Self-report assessments of trait coping are poor measures of the trait component of momentary coping and very poor predictors of coping in specific situations. PMID- 10474212 TI - The impact of personality on the reporting of unfounded symptoms and illness. AB - This study examined the role of personality in the reporting of symptoms and illness not supported by underlying pathology. After assessment of the Big Five personality factors, 276 healthy volunteers were inoculated with a common cold virus. On each of the following 5 days, objective indicators of pathology, self reported symptoms, and self-reported illness onset were assessed. Neuroticism was directly associated with reports of unfounded (without a physiological basis) symptoms in individuals at baseline and postinoculation in those with and without colds. Neuroticism was also indirectly associated with reports of unfounded illness through reports of more symptoms. Openness to Experience was associated with reporting unfounded symptoms in those with verifiable colds, whereas Conscientiousness was associated with reporting unfounded illness in those who were not ill. PMID- 10474213 TI - Quality of early family relationships and individual differences in the timing of pubertal maturation in girls: a longitudinal test of an evolutionary model. AB - In an 8-year prospective study of 173 girls and their families, the authors tested predictions from J. Belsky, L. Steinberg, and P. Draper's (1991) evolutionary model of individual differences in pubertal timing. This model suggests that more negative-coercive (or less positive-harmonious) family relationships in early childhood provoke earlier reproductive development in adolescence. Consistent with the model, fathers' presence in the home, more time spent by fathers in child care, greater supportiveness in the parental dyad, more father-daughter affection, and more mother-daughter affection, as assessed prior to kindergarten, each predicted later pubertal timing by daughters in 7th grade. The positive dimension of family relationships, rather than the negative dimension, accounted for these relations. In total, the quality of fathers' investment in the family emerged as the most important feature of the proximal family environment relative to daughters' pubertal timing. PMID- 10474214 TI - When ideology hurts: effects of belief in the protestant ethic and feeling overweight on the psychological well-being of women. AB - Two studies addressed the relationship between Protestant ethic (PE) ideology and psychological well-being for self-perceived overweight and normal weight women. In Study 1, PE beliefs interacted with self-perceived weight status: For very overweight women, higher PE beliefs were related to lower psychological well being, whereas the opposite pattern emerged for normal weight women. The relationship of PE to well-being was not mediated by beliefs about controllability of weight or dislike of the overweight. In Study 2, either a PE ideology or an inclusive ideology was primed within the context of the stigma of overweight. For overweight participants, priming PE ideology led to decreased psychological well-being, whereas priming an inclusive ideology led to increased psychological well-being. Normal weight participants were unaffected. PE ideology as a vulnerability factor for the psychological well-being of the overweight is discussed. PMID- 10474215 TI - Making drugs out of oligonucleotides: a brief review and perspective. AB - I provide a brief review and perspective thoughts concerning the antisense oligonucleotide, drug discovery paradigm. PMID- 10474216 TI - The 9-fluorenylmethoxycarbonyl (Fmoc) group and its use in oligonucleotide synthesis. AB - The introduction of the base-labile 9-fluorenylmethoxycarbonyl (Fmoc) group into the exocyclic amino function of 2'-deoxynucleosides and their dimethoxytritylation and phosphitylation is described. The resulting key intermediates were investigated in the built-up of different oligodeoxyribonucleoside phosphate and thiophosphate chains which were deprotected under mild basic conditions leading to crude oligomers of high purity. PMID- 10474217 TI - Antiviral activity of magnesium and magnesium/poly r(A-U) combinations against two RNA viruses. AB - Magnesium (Mg2+) potentiated the anti-vesicular stomatitis virus (VSV) activity of poly r(A-U) or poly r(G-C) and the anti-HIV-1 activity of poly r(A-U). Mg2+ did not affect the anti-VSV activity of poly (rI).poly (rC), poly (dA-dT).poly (dA-dT) or poly (dG-dC).poly (dG-dC). Modulation of one or more nuclear (nucleolar) processes of the host cell may be responsible for the synergistic antiviral activity. PMID- 10474218 TI - High-density nucleoside analog probe arrays for enhanced hybridization. AB - DNA probe arrays were synthesized with analogs of 2,6-diaminopurine and 2'-O methyl-thymidine in place of A and T. AT-rich GeneChip test arrays containing 14 mer or 20-mer analog probes improved hybridization to fluorescently-labeled RNA sequences under stringent conditions. PMID- 10474219 TI - Detection of nucleic acids by cycling probe technology on magnetic particles: high sensitivity and ease of separation. AB - Cycling Probe Technology (CPT) is a signal amplification system that allows detection of nucleic acid target sequences without target amplification. CPT employs a sequence specific chimeric probe, typically DNA-RNA-DNA, which hybridizes to a complementary target DNA sequence and becomes a substrate for RNase H. Cleavage occurs at the RNA internucleotide linkages and results in dissociation of the probe from the target, thereby making it available for the next probe molecule. This communication describes the use of oligonucleotides attached to solid supports for target capture and release followed by solution and solid phase cycling. Through the attachment of chimeric probes to Sera-Mag magnetic particles (SMP) a simple and effective method of separating the cleaved probe from non-cycled probe has been developed. By capturing the target DNA on particles and separating it from the extraneous non-specific DNA we are able to dramatically reduce background and thus discriminate between samples of Methicillin Resistant (MRSA) and Methicillin Sensitive (MSSA) Staphylococcus Aureus. We conjugated oligonucleotide probes to SMPs (approximately 1 um) and Nylon beads (NB) which were coated with ID Biomedical's proprietary coating materials (R, patent pending). The general structure of the constructs is shown below: [table: see text] PMID- 10474220 TI - Solid-supported oligonucleotide systems for special biomedical applications. AB - The use of composite beads consisting of a 6 microns polystyrene core with 30 nm surface-bound silica particles to routine automatic oligodeoxynucleotide (ODN) synthesis is described. PMID- 10474221 TI - Chemical synthesis of oligonucleotides containing the (6-4) photoproduct at the thymine-cytosine site and its repair by (6-4) photolyase. AB - A phosphoramidite building block of the T(6-4)C photoproduct was synthesized. One of the differences from T(6-4)T was formation of cytosine hydrates by UV irradiation, and the other was acylation of the amino function with the capping reagent. The capping step was omitted to improve the yield of the desired oligonucleotides. Characterization of the (6-4) photolyase using one of the oligonucleotides revealed that this enzyme restores the pyrimidines in T(6-4)C to their original structures. PMID- 10474222 TI - Synthesis and biological evaluation of modified DNA fragments for the study of nucleotide excision repair in E. coli. AB - Three new cholesterol-containing phosphoramidites where synthesized and used in automated synthesis of modified DNA fragments. These cholesterol lesions are good substrates for the E. coli UvrABC endonuclease. In vitro they are incised from damaged DNA with higher efficiency in respect with the cholesterol lesions previously published. PMID- 10474223 TI - Boranophosphate nucleic acids--a versatile DNA backbone. AB - Important chemical and biochemical properties of boranophosphate DNA and RNA oligonucleotides are reviewed. Stereoregular boranophosphate oligomers can be synthesized enzymatically and form stable duplexes with DNA. Fully boronated, non stereoregular oligothymidylates, synthesized chemically, form hybrids with poly(A) that have lower melting points than oligothymidylate:poly(A), yet they nevertheless can support the RNase H mediated cleavage of RNA. PMID- 10474224 TI - MMI linkage modification increases potency and stability of H-ras antisense oligonucleotides. PMID- 10474225 TI - Interaction of complementary oligonucleotides with the 3'-end of yeast tRNA(Phe). AB - Interaction of yeast tRNA(Phe) with oligodeoxyribonucleotides (ONs), complementary to the nucleotides 62-76 was investigated. Results of gel-mobility shift assay and RNase A probing evidence that the ONs containing the sequence complementary to the tRNA ACCA end can easily invade the hairpin structure under physiological conditions. The limiting step of association process is the tRNA unfolding. PMID- 10474226 TI - Mechanism and specificity of RNA cleavage by chemical ribonucleases. AB - Cleaving of model RNA substrates by chemical ribonucleases constructed by conjugation of 1,4 diazabicyclo[2,2,2]octane with histamine and histidine was investigated. Similarly to RNase A, the chemical RNases produce fragments with 5' hydroxy-group and 3'-cyclophosphate. The cleavage occurs as the catalytic reaction: more than 150 phosphodiester bonds in RNA can be cleaved by one molecule of RNase mimic. PMID- 10474227 TI - Tc-99m-labeling of modified RNA. AB - The synthesis of Tc-99m-labeled, modified RNA is reported. This new class of radiopharmaceuticals is of potential interest as target specific imaging agents. The preparation of N3S-RNA was achieved by coupling protected MAG2-units to amino modified ON's. The N3S-RNA was Tc-99m-labeled with 90-95% radiochemical yield and specific activities of 37MBq/nmol leading to 1:1-Tc-99m-N3S-aptamers. PMID- 10474228 TI - Synthesis and biological activity of 2',5'-oligoadenylate trimers containing 5' terminal 5'-amino-5'-deoxy- and 5'-amino-3',5'-dideoxyadenosine derivatives. AB - Some new 2',5'-oligonucleotide trimers containing 5'-terminal 5'-amino-5'-deoxy- and 5'-amino-3',5'-dideoxyadenosine derivatives have been synthesized. Some of the trimers showed biological inhibitions of HIV-1 reverse transcriptase (RT), HIV-1 induced syncytia formation and PCR amplification. PMID- 10474229 TI - 2-5A-PNA complexes: a novel class of antisense compounds. AB - This paper presents the fully automated solid phase synthesis of 2-5A-PNA hybrids. These stable antisense probes cause RNase L mediated hydrolysis of target RNA sequences. PMID- 10474230 TI - Synthesis of new photocross-linking 5-C-base-substituted UTP analogs and their application in highly selective affinity labelling of the tick-borne encephalitis virus RNA replicase proteins. AB - A new photocross-linking 5-C-base-substituted UTP analogs, carrying 4 azidoperfluorobenzoyl and 4-azidoaniline residues were synthesized. Two flavivirus proteins NS5 and NS3 are shown to be labelled after RNA synthesis in the presence of the analogs, irradiation (lambda > 300 nm) and subsequent [alpha 32P]NTP incorporation. PMID- 10474231 TI - Using fluorescence resonance energy transfer (FRET) for measuring 2-5A analogues ability to activate RNase L. AB - The development of a method for measuring the ability of 2-5A analogues to activate the cleavage of an oligoribonucleotide substrate by RNase L is described. This method is based on fluorescence resonance energy transfer. The method is easily performed with 96-well plates, allowing for quantitative high throughput analyses of 2-5A analogues under different reaction conditions. PMID- 10474232 TI - New techniques for the rapid characterization of oligonucleotides by mass spectrometry. AB - Recent advances in combined HPLC/electrospray ionization-mass spectrometry provide effective new capabilities for the rapid characterization of oligonucleotides. Accurate mass measurements with errors < 0.3 Da, and determination of base and sugar modification and of nearest neighbor identities, can be routinely carried out on 10-100 component mixtures of RNA or DNA. These procedures are widely applicable in structural and analytical studies involving mixtures of oligonucleotides. PMID- 10474233 TI - Thermodynamics of site-specific variant tRNA(Ala) acceptor stem microhairpins. AB - Thermal denaturation studies were carried out on a set of site-specific variants of a 22mer RNA hairpin comprising the aminoacyl acceptor stem sequence of E. coli tRNA(Ala). The pairing thermodynamics were calculated from the melting profiles. PMID- 10474234 TI - Cytarabine-induced destabilization and bending of a model Okazaki fragment. AB - The effects of cytarabine on the structural and thermodynamic properties of an Okazaki fragment were investigated using UV hyperchromicity and 2D 1H NMR. Cytarabine significantly decreased the stability of this model Okazaki fragment, decreasing the melting temperature from 46.8 degrees C to 42.4 degrees C at 1.33 x 10(-5) M. Cytarabine also markedly increased the bend angle of the Okazaki fragment duplex from 20 degrees to 42 degrees. Changes to the structures and stabilities of Okazaki fragments may cause the biological effects of cytarabine. PMID- 10474235 TI - Nucleoside modifications affect the structure and stability of the anticodon of tRNA(Lys,3). AB - NMR spectroscopy was used to determine the solution structures of RNA oligonucleotides comprising the anticodon domain of tRNA(Lys,3). The structural effects of the pseudouridine modification at position 39 were investigated and are well correlated with changes in thermodynamic parameters. The loop conformation differs from that seen in tRNA(Phe) and provides an explanation of the critical role of modification in this tRNA. PMID- 10474236 TI - Formation of triple helices at irregular poly (R.Y) sites located in critical positions in the human bcr promoter. AB - Two polypurine sequences interrupted respectively by one and two adjacent pyrimidines have been identified in the promoter of the human bcr gene. Although these targets are irregular they are recognised and tightly bound by AG and GT motif triplex-forming oligonucleotides. Thermodynamic and kinetic data are presented. PMID- 10474237 TI - A single carbocyclic nucleotide substitution in a 12mer DNA gives a Hoogsteen basepaired duplex (till 38 degrees C) and a hairpin (till 65 degrees C). A 600 MHz NMR spectroscopic study. AB - The impact of intramolecular stereoelectronic effects has been examined by comparison of the solution structures of natural oligo-DNA duplex, 5'(1C2G3C4G5A6A7T8T9C10G11C12G)2(3'), and its carbocyclic-nucleotide analogues in which the pentose sugar in 2'-dA residue is replaced with its carbocyclic counterpart (i.e. 2'-deoxyaristeromycin). Based on the NMR evidences, it has been shown, that 2'-deoxyaristeromycin analog exists in a dynamic equilibrium between the two forms of duplexes, one with W-C bp and the second with Hoogsteen bp in ca 1:1 ratio at lower temperature (below 35 degrees C) and as hairpin at higher temperature (from approximately 40 degrees-60 degrees C). PMID- 10474238 TI - Sequence-specific alkylation of DNA by duocarmycin A and its novel derivatives bearing PY/IM polyamides. AB - A new class of sequence-specific DNA alkylating agents were developed based on the reactivity of duocarmycin A and the DNA-reading ability of pyrrole-imidazole polyamide. The DNA alkylation sequence specificity by duocarmycin A can be modulated by a variety of pyrrole-imidazole triamides in a predictable manner. Novel hybrids of the segment A of duocarmycin A and pyrrole-imidazole polyamides efficiently and highly selectively alkylated the target base possessing match sequences of Dervan's binding code. PMID- 10474239 TI - Synthesis of oligonucleotide-intercalator conjugates capable to inhibit HIV-1 DNA integration. AB - This investigation is devoted to design of short "switch" oligonucleotides mono- or bi-functionnalized with intercalating agents capable to form a stable triplex with HIV integrase-cognate sequences and inhibit selectively HIV integration. Methods of intercalator incorporation at 5'- and/or 3'-terminal positions or one of the pyrimidine heterocyclic bases are developed. PMID- 10474240 TI - Binding of chemically-modified oligonucleotides to the double-stranded stem of an RNA hairpin. AB - We monitored the binding of triplex-forming oligopyrimidines to the double stranded stem of the RNA hairpin responsible for the gag-pol frameshift in HIV-1. Whereas the substitution of 5, propynyl-C for C had a limited effect, the use of a Peptide Nucleic Acid 12mer led to a drastic reduction in the stability of the oligomer/RNA complex. PMID- 10474241 TI - Relative effects of graft copolymer and polyamines on triplex stabilization under physiological conditions. AB - Triplex-stabilizing effect of a graft copolymer under physiologically relevant conditions has been evaluated and compared with other polyamines. Here we show that the graft copolymer significantly stabilizes triplex DNAs with amazingly higher efficacy than that of physiological concentrations of spermine and spermidine. PMID- 10474242 TI - Enzymatic hydrolysis and biological activity of oligonucleotides containing 5 substituted pyrimidine bases. AB - Two series of alternating ODNs containing 5-n.alkyl-, alkenyl- and alkynyl-dU and -dC units have been prepared in order to study the kinetics of their hydrolysis by SV PDE and human serum, respectively. Both in (r5dUpdA)10 and (r5dCpdG)6 series the rate of hydrolysis decreased with increasing length of side-chain. Replacement of thymidines by 5-hexynyl-dU in different antisense oligomers resulted in considerably higher biological activity relative to that of the thymidine-containing counterparts. PMID- 10474243 TI - A 38 kDa nuclear protein is involved in the retention of an antisense oligonucleotide directed against cytosolic phospholipase A2. AB - Recent studies suggest that antisense phosphorothioate oligonucleotides (APO) are useful tools not only to impair gene expression, but also to modify the splicing of pre-mRNA, as the classical view that they act by suppressing the translation of mature mRNA has been challenged by several examples showing their nuclear site of action. In this work we show that an APO directed against cytosolic phospholipase A2 (cPLA2) mRNA localises in the nucleus and interacts with a specific nuclear protein. PMID- 10474244 TI - Specifically designed polymeric nanospheres increase cellular uptake of unmodified antisense ODNs. AB - The cellular uptake and the inhibitory effect of c-myb unmodified antisense oligonucleotides reversibly bound to new polymeric nanoparticles in HL-60 cellular system have been found to increase by 50 folds if compared with the free ODN. An initial single dose (320 nM) of the nanoparticle bound unmodified antimyb ODN has been able to specifically inhibit HL-60 leukemia cell proliferation for at least 8 days. PMID- 10474245 TI - Comb-type copolymers for controlled DNA delivery. AB - Various comb-type copolymer containing a polycation as a main chain was design to construct delivery systems of DNAs. The comb-type copolymers having cell-specific polysaccharides were proved to be useful to deliver DNA to the target cells in vivo. Of interest, the copolymers with abundant side chains of hydrophilic polymers are capable of stabilizing DNA triplex. Further, injectable nanoparticles for controlled releases of DNAs were fabricated from the copolymer and a biodegradable polymer. PMID- 10474246 TI - In vitro and in vivo anti-influenza A virus activity of antisense oligonucleotides. AB - We have demonstrated that antisense phosphorothioate oligonucleotides (S-ODNs) inhibit influenza virus A replication in MDCK cells. The liposomally encapsulated and the free antisense phosphorothioate oligonucleotides with four target sites (PB1, PB2, PA, and NP) were tested for their abilities to inhibit virus-induced cytopathogenic effects by a MTT assay using MDCK cells. The liposomally encapsulated S-ODN complementary to the sites of the PB2-AUG initiation codon showed highly inhibitory effects. Therefore, the antiviral effects of S-ODN-PB2 AUG and PA-AUG were examined in a mouse model of influenza virus A infection. PB2 AUG oligomer treated i.v. significantly prolonged the mean survival time in day (MDS) and increased the survival rates with does dependent manner. PMID- 10474247 TI - Thermodynamic studies on hairpin forming antisense-oligodeoxynucleotides directed against hepatitis C virus RNA. AB - Phosphorothioate and benzyl-modified antisense-oligodeoxynucleotides directed against nucleotides 334-350 of the Hepatitis C Virus RNA form surprisingly stable hairpins. These data contribute to solve a structural detail information in search for a global secondary structure model of the Non Coding Region (NCR) of HCV. PMID- 10474248 TI - Activation of spleen lymphocytes by plasmid DNA. AB - It was shown that plasmid pUC19 DNA stimulates in vitro proliferation of CBA mouse splenocytes in a dose-dependent manner. Stimulation effect of the plasmid DNA is additive with COn A or LPS, synergistic with PMA and is inhibited by nonimmunogenic phosphodiester oligonucleotides and Fab fragments of antimouse Ig antibodies. These data and the data of affinity labelling of ODN-binding proteins indicate that immunoglobulin receptors are involved in DNA-induced lymphocyte activation. PMID- 10474249 TI - Uptake of oligonucleotides by keratinocytes. AB - Oligonucleotides (ODNs) conjugated to rhodamin (Rh) and 4-[(N-2-chloroethyl-N methyl)amino] benzylamine were used to investigate ODNs transport into keratinocytes. Affinity labeling of two proteins, 63 and 35 kDa, and the inhibition of the affinity labeling and ODNs uptake by the cells in the presence of nucleic acids, polyanions and trypsin suggest, that the proteins are involved in transport of nucleic acids in keratinocytes. PMID- 10474250 TI - Antisense effects of oligonucleotides complementary to the hairpin of the Leishmania mini-exon RNA. AB - We investigated the binding and the translation inhibitory properties of hexadecamers complementary to the mini-exon sequence of the protozoan parasite Leishmania amazonensis. This targeted RNA region folds into a hairpin. Large differences were observed in the antisense properties of the different oligomers although their binding to RNA always requires the disruption of the stem region. PMID- 10474251 TI - Antisense oligodeoxynucleotide complementary to CXCR4 mRNA block replication of HIV-1 in COS cells. AB - CXCR4 is both a chemokine receptor and an entry co-receptor for the T-cell line adapted human immunodeficiency virus type 1 (HIV-1). To find a more efficacious therapeutic treatment of acquired immunodeficiency syndrome, we examined the effects of antisense oligonucleotides on CXCR4 production. COS cells, stably expressing CXCR4 and CD4, were incubated with several kinds of oligonucleotides. Total human p24 antigen production was determined using an enzyme-linked immunosorbent assay system. An antisense phosphorothioate-modified oligonucleotide, complementary to the translation initiation region of the CXCR4 mRNA, showed minimal inhibition of p24 antigen production at the high concentration of 2 microM. On the other hand, the antisense phosphorothioate oligonucleotide, when used with transfection reagents, showed high efficiency at low concentrations, and confirmed the sequence-specific action. Interestingly, the oligonucleotide with the natural phosphodiester backbone, when used with the transfection reagents, also had high functional effects, comparable to the modified oligonucleotide. This study defines the prerequisite criteria necessary for the design and the application of antisense oligonucleotides against HIV-1 in vivo. PMID- 10474252 TI - The cytotoxicity of anti-PAI-1 oligonucleotides and their conjugates. AB - The cytotoxicity of anti-PAI-5 hexadecanucleotides (phosphodiesters and phosphorothioates) and their conjugates with lipophilic alcohols was tested in EA.hy 926 hybrid endothelial cells. Some cytotoxicity was found for cholesteryl and bornyl conjugates at concentrations higher than those used for antisense inhibition experiments. PMID- 10474253 TI - Effect of oligomer length and base substitutions on the cytotoxic activity and specific nuclear protein recognition of GTn oligonucleotides in the human leukemic CCRF-CEM cell line. AB - We have identified phosphodiester oligonucleotides composed of G and T bases, named GTn, which are able to inhibit the cellular growth of human cancer cell lines by recognising specific nuclear proteins. We demonstrated that GTn oligonucleotides require a length of at least 20 nucleotides in order to exert a significant cytotoxic effect and to retain the specific protein binding ability. In addition, we found that GTn cytotoxicity was lost when A or C bases were introduced at either 3' and 5' end or within the GTn sequences. PMID- 10474254 TI - Inhibition of HIV-1 integration by mono- & bi-functionalized triple helix forming oligonucleotides. AB - HIV-1 DNA integration is carried out by integrase, a viral protein which binds to specific sequences located on both extremities of the HIV-1 DNA LTR. Inhibition of integration was observed with submicromolar concentrations of mono- or bifunctionalized 11-mer oligonucleotide-intercalators, which were designed to form an alternate strand triple helix with the U5 LTR end containing two adjacent purine tracts on opposite strands 5'-GGAAAATCTCT-3'/3'-CCTTTTAGAGA-5'. PMID- 10474255 TI - Protein and RNA of human telomerase as targets for modified oligonucleotides. AB - Chimeric oligodeoxynucleotides containing phosphorothioate and N3'-->P5' phosphoramidate linkages were synthesized. These oligomers show a high inhibitory activity against human telomerase. PMID- 10474256 TI - Uptake and intracellular distribution of oligonucleotides vectorized by a PAMAM dendrimer. AB - We studied the uptake and intracellular distribution of an FITC labelled phosphodiester oligodeoxynucleotide (ODN) vectorized by a dendrimeric structure in cell culture. PMID- 10474257 TI - Cytotoxicity and in-vivo tolerance of FdUMP[10]: a novel pro-drug of the TS inhibitory nucleotide FdUMP. AB - The cytotoxicity of the 10mer ODN FdUMP[10] towards human colorectal tumor cells was evaluated using a clonogenic assay. FdUMP[10] was more than 100-fold more active than 5-FU at inhibiting colony formation of H630 cells. FdUMP[10] was also evaluated for cytotoxicity in the NCI 60 cell line screen, and showed markedly improved activity relative to 5-FU against numerous tumor cell lines. The in-vivo tolerance of FdUMP[10] is more than three-fold greater per mole fluorinated pyrimidine, than 5-FU. PMID- 10474258 TI - Synthesis of modified thiopurine nucleosides for structural characterization of human thiopurine S-methyltransferase. AB - Synthesis of a number of photoactive thiopurine-containing nucleosides was described. S-methylation of the synthesized compounds in the course of the reaction catalyzed by recombinant human thiopurine S-methyltransferase was studied by UV-spectroscopy. PMID- 10474259 TI - Dendroanalysis: a tool for biomonitoring environmental pollution? AB - Trees of temperate regions usually form visible annual growth rings, which can be dated accurately. It is therefore possible to collect wood samples of different age and analyse their heavy metals content in order to get a chronological record of trace elements pollution in the tree's environment. This method of retrospective biomonitoring was called dendroanalysis. A basic assumption of dendroanalysis is the stability of the mineral distribution patterns, i.e. once the elements are stored, no significant mobility should occur. Additionally, neighbouring trees growing in the same environment should show similar radial element patterns. While some studies presented good correlations between radial distributions of heavy metals in tree rings and temporal records of pollution from industry or traffic, others failed in using dendroanalysis as a chronological record of pollution. Probably some elements can move at a certain rate in radial direction through the ray parenchyma cells. In this way the radial element distributions are subsequently changed. Growth rates of tree rings can also influence the concentrations of elements in wood. During periods of slow growth higher concentrations of elements can be found in the wood. Therefore, radial distribution patterns of heavy metals in tree rings should be used with caution as a tool for retrospective biomonitoring of environmental pollution. PMID- 10474260 TI - Biomonitoring of airborne inorganic and organic pollutants by means of pine tree barks. I. Temporal and spatial variations. AB - Scots pine (Pinus sylvestris L.) bark samples were collected at two field sites (Neuglobsow, Rosa) and in different years between 1987 and 1996 in the east of Germany. The barks were analyzed with respect to the following inorganic and organic substances: Al, As, B, Ca, Cd, Ce, Cr, Cu, Fe, Hg, Mo, NH4+, Ni, NO3-, PO4(3)-, Pb, Sr, SO4(2)-, Ti, V, W, Zr, Zn, benzo[a]pyrene, fluoranthene, pyrene, alpha-hexachlorocyclohexane (alpha-HCH) and dichlorodiphenyltrichloroethane (DDT). In addition to bark samples from the site Rosa, 53 test sites were investigated in the Nature Park Dubener Heide. Here, the analysis of the barks aimed at discovering spatial patterns of the above-mentioned substances. Since 1991, most of the determined substances (e.g. sulfate, nitrate, calcium, lead, benzo[a]pyrene, alpha-HCH) show decreased concentration values in bark samples from both sites. Temporal variations reflect substantial infra-structural changes in eastern Germany, especially at Rosa and in the industrial region around the cities Leipzig, Halle, and Bitterfeld. Moreover, nitrate concentrations in barks are increasing since 1995. The trend can be explained with increased nitrogen emissions from motor traffic and livestock farms. Spatial patterns of sulphate and ammonia reflect inputs from power plants and agriculture in pine stands of the Nature Park Dubener Heide. The results show that barks of pine trees can be used as biomonitoring tools to indicate and characterize depositions of airborne organic and inorganic pollutants. PMID- 10474261 TI - Large scale air monitoring: lichen vs. air particulate matter analysis. AB - Biological indicator organisms have been widely used for monitoring and banking purposes for many years. Although the complexity of the interactions between organisms and their environment is generally not easily comprehensible, environmental quality assessment using the bioindicator approach offers some convincing advantages compared to direct analysis of soil, water, or air. Measurement of air particulates is restricted to experienced laboratories with access to expensive sampling equipment. Additionally, the amount of material collected generally is just enough for one determination per sampling and no multidimensional characterization might be possible. Further, fluctuations in air masses have a pronounced effect on the results from air filter sampling. Combining the integrating property of bioindicators with the world wide availability and particular matrix characteristics of air particulate matter as a prerequisite for global monitoring of air pollution is discussed. A new approach for sampling urban dust using large volume filtering devices installed in air conditioners of large hotel buildings is assessed. A first experiment was initiated to collect air particulates (300-500 g each) from a number of hotels during a period of 3-4 months by successive vacuum cleaning of used inlet filters from high volume air conditioning installations reflecting average concentrations per 3 months in different large cities. This approach is expected to be upgraded and applied for global monitoring. Highly positive correlated elements were found in lichens such as K/S, Zn/P, the rare earth elements (REE) and a significant negative correlation between Hg and Cu was observed in these samples. The ratio of concentrations of elements in dust and Usnea spp. is highest for Cz, Zn and Fe (400-200) and lowest for elements such as Ca, Rb, and Sr (20-10). PMID- 10474262 TI - Improving the use of lichens as biomonitors of atmospheric metal pollution. AB - The data reported on this study supported the hypothesis that the absence of the lichen Ramalina fastigiata near a copper mine site located on the south of Portugal was related to toxic levels of Cu-dust near the centre of the mine. Lichen biodiversity reflected the impact of the copper-mine dust emissions which were more widespread towards the east, correlated with wind direction and frequency. The chemical analysis of R. fastigiata collected at different distances and in different directions from the mine showed that Cu, K and Mg were derived from the centre of the mine site, confirming thus as the major source of atmospheric dust. Total inhibition of PSII photochemical reactions occurred in R. fastigiata both under field and controlled conditions, when intracellular Cu concentrations exceeded a threshold of approximately 2.0 mumol g-1. No samples of this species were found under field conditions beyond the Cu threshold. Thus, the fluorescence parameter Fv/Fm proved to be a good estimator of the survival capacity of R. fastigiata under field conditions and thus a useful parameter in determining the sensitivity of the lichens (photobiont) to Cu pollution. The intracellular location of Cu allowed an explanation of the physiological changes and the survival of the species in the surroundings of the copper-mine. PMID- 10474263 TI - [Controversies and current status of therapy of optic nerve damage in craniofacial traumatology and surgery]. AB - In craniomaxillofacial traumatology, surgical oncology and craniomaxillofacial reconstruction, a surgeon's aim may interfere with the prechiasmatic visual pathway. Precise concepts and therapeutic strategies are mandatory to detect and deal with anterior visual pathway disorders. In order to develop these strategies, knowledge of the pathomechanisms of potential optic nerve trauma, primary radiological investigations, and further diagnostic measures are important. Due to the difficulties in neuroophthalmological testing of visual pathway functioning in severely injured patients or even during craniomaxillofacial reconstructions, we established flash-evoked visual potentials (VEP) and the electroretinogram (ERG) as reliable electrophysiological methods to gather specific information as to whether the visual pathway function is intact, even if pathological, but still present or absent. Case reports show that subjectively or objectively confirmed unilateral amaurosis does not necessarily mean irreversible vision loss. The electrophysiological evaluation together with multiplanar computer tomography (CT) are important for the immediate identification of optic nerve trauma. The results of this evaluation will provide the diagnostic information on whether surgical intervention and/or conservative therapy is required to prevent secondary optic nerve damage. The conservative therapy of choice for the treatment of traumatic optic nerve lesions is the methylprednisolone-megadosis regimen (30 mg Urbason/kg bodyweight i.v. and 5.4 mg/kg bodyweight/h i.v. for the following 47 h). Surgical therapy involves decompression of the orbital compartment in case of retrobulbar hematoma or decompression of the intracanalicular part of the optic nerve in the traumatized optic canal or posterior orbit as confirmed by CT. Prospective analysis of our trauma patients and the international literature on traumatic optic nerve lesions show that the time factor in when to start therapy has been greatly underestimated. To fulfill modern treatment concepts in craniomaxillofacial surgery, sound diagnostic and therapeutic knowledge on the maintenance of visual pathway function is required. PMID- 10474264 TI - [Facial cleft birth rate in former East Germany before and after the reactor accident in Chernobyl]. AB - Cleft lip palates (CLP) are caused by a variety of factors. Ionizing radiation is only one of these factors. The meltdown of the nuclear reactor at Chernobyl on April 26, 1986, and the subsequent radioactive fallout did not cause any acute radiation sickness in Germany. Nevertheless, in West Berlin a significant increase of trisomy-21 cases was reported in births 9 months after the Chernobyl reactor accident. In our study we analyzed the influence of the radioactive fallout after the Chernobyl disaster on the rate and regional distribution of CLP newborns in the former German Democratic Republic (GDR). In contrast to the Federal Republic of Germany an ongoing malformation register for CLP newborns had existed in the former GDR since 4 July 1967. Environmental data were collected from national and international environmental authorities and atomic energy agencies. Population statistics were taken from the statistical year-book of the former GDR. During a 10-year period from 1980 to 1989, the average number of CLP newborns in the GDR was 1.88 per 1,000 live births. A significant prevalence increase was recorded in 1983, 1987 und 1988. In comparison to the mean rate in the period from 1980 until 1986, 1987 demonstrated an increase of 9.4%. Regional prevalence increases were seen in the three northern districts of Schwerin, Rostock and Neubrandenburg, where the radioactivity measurements in general showed higher levels of the radionuclides caesium-137 und strontium-90 than in other districts. Owing to the comprehensive malformation register for CLP patients in the GDR, this is the first study for Germany, analyzing the CLP rate before and after the fallout in Chernobyl. The results support the allegation of the influence of radiation-induced increase of CLP newborns after the Chernobyl reactor accident. PMID- 10474265 TI - [Tissue damage and regeneration after laser surgery using different scanning systems]. AB - INTRODUCTION: Laser systems are frequently used in dentistry. New laser scanner technologies are promising more homogeneous ablation of pathological ablations of the skin and mucosa. The theoretical advantages of these systems have not yet been sufficiently evaluated by histological findings. For this reason, we compared two laser scanners with different scanning patterns in this study (Silktouch and Swiftlase, by Sharplan, Germany). MATERIALS AND METHODS: In this animal study (79 male Osborn-Mendel rats), skin defects of 3-mm diameter were lasered on to the backs of the animals, one for each method mentioned above (defocused laser, Swiftlase, and Silktouch). The histological investigations were conducted, depending on the time after surgery (9 h, 1, 2, 3, 4, 5, 6, 7, 9, 11, or 13 days). On the clinical site, we evaluated the visible ablation rate, homogeneity and morphology of the wound surface and the degree of carbonization. In the histomorphological investigations, we determined the degree of thermal damage as well as the morphology of the necrotic area and reepithelization pattern of the surface. RESULTS: The Silktouch scanner clinically showed a homogeneous ablation rate with less carbonization than the Swiftlase scanner. In the histological specimen, this correlated with an increase in the tissue ablation rate and decreasing thermal damage to the surrounding tissue. Especially in the early wound-healing period, there was a more rapid reepithelization after the use of the Silktouch scanner. These differences could not be evaluated in the later wound-healing period. Compared to the defocused laser ablation, both scanning systems seem to have benefits within these parameters. CONCLUSIONS: The results of this study seem to show a diminished perifocal damage and reduced clinical postoperative morbidity achieved by using laser scanning systems. Compared to the Swiftlase laser, the Silktouch seems to deliver better results. PMID- 10474266 TI - [Visualizing carcinomas of the mouth cavity by stimulating synthesis of fluorescent protoporphyrin IX]. AB - The fluorescence diagnosis based on the aminolavulinic acid-stimulated porphyrin synthesis allows the detection of superficial tumors in a very early stage even when they are very small tumors. A fluorescence diagnosis of tumors in the oral cavity can be simply performed by rinsing with a 0.4% ALA solution for 20 min. This topical application avoids systemic side effects such as skin sensitization. The red fluorescent areas are visible to the naked eye; only a blue light source for fluorescence excitation is necessary and a suitable red filter for observation. In our study on 56 patients suffering from carcinoma of the oral cavity, 96% of the histologically confirmed carcinoma could be visualized via fluorescence. In 3 patients additional tumors were detected via fluorescence that were not visible otherwise. However, many patients show fluorescent areas with no correlation to the histological finding. It was verified that bacteria from the oral cavity also produce PpIX after ALA incubation, which leads to false-positive findings. Reduction of the false-positive findings was achieved by rinsing the oral cavity with hydrogen peroxide and by mechanical plaque reduction. This improves the reliability of a fluorescence-guided biopsy. However, suppression of the bacteria fluorescence is necessary for clinical use of this diagnostic method. PMID- 10474267 TI - [Inhibition of tumor neo-angiogenesis and induction of apoptosis as properties of docetaxel (taxotere)]. AB - Paclitaxel and docetaxel are potent drugs that are effective in the treatment of malignant tumors. The cytotoxic action of these drugs is not fully understood, but it appears to be mediated mainly through mitotic arrest and subsequent apoptosis. Because no information is available on the antiangiogenesis action of docetaxel, the investigations were performed to determine whether inhibition of neoangiogenesis plays a role in docetaxel's antitumor efficacy. Four different mouse tumors, two squamous cell carcinomas (SCC-IV; SCC-VII) and two adenocarcinomas (MCA-4; MCA-29) were assayed for angiogenic activity using the in vivo i.c. angiogenesis assay. Tumor cells (5 x 10(5)) were injected i.c. into the skin flap over the abdominal wall, and the number of new blood vessels at the tumor cell injection site was determined 2, 4, 6, 8, 10 and 12 days later. The mice were treated with docetaxel (Taxotere--31.3 mg/kg i.v.) 1 or 4 days after tumor cell injection. The number of new blood vessels increased with time. Docetaxel reduced the number of newly formed blood vessels in MCAs, but not in SCCs. The reduction was associated with slower tumor growth. In a separate set of experiments we observed that docetaxel's inhibitory effect on the two MCAs was histologically associated with massive tumor cell destruction by means of both apoptosis and necrosis. This was not observed for the two SCCs. Since no reduction in blood vessels occurred in tumors unresponsive to docetaxel, the inhibition of neoangiogenesis in docetaxel-responsive tumors was likely the result of a decrease in angiogenic stimuli due to docetaxel's destruction of tumor cells. PMID- 10474268 TI - [Anterior and lateral disk and capsule tissue of the temporomandibular joint and its relevance for temporomandibular joint diagnosis]. AB - OBJECTIVE: Fibrocartilage that bears tensile stress is typically fixed bilaterally to adjacent bone by collagenous fibres. For the temporomandibular joint, such disc fixations in the anterior and lateral regions have not been described, even in MRT findings of anterior disc dislocations without reduction. By observing collagenous fibres of the anterior and lateral discuss and capsule tissue, we sought to establish whether tensile load capacities typical for fibrocartilage also apply to the discuss articularis of the mandibular joint in sagittal direction. MATERIALS AND METHODS: Using dissection material from 20 adult and 10 newborn temporomandibular joint specimens, the position of the caput mandibulae in the fossa mandibularis or below the tuberculum articulare was ascertained. Serial sections were stained using alternately haemotoxylineosin and the Goldner method. RESULTS: Collagenous fibres from the medial and lateral discuss insert into the corresponding caput pole and form the limit of the lower joint cavity. In the anterior capsule region, two fibres may be seen coming in similar fashion from the pes disci to the retrodiscal bilaminary zone. They are denoted respectively as inferior and superior stratum anterior. They limit the upper and lower joint cavity in anterior direction. Depending on motion, both experience various tensile loads, together with the inferior and superior stratum posterior. The tensile loads exerted are described and clarified with drawings. CONCLUSION: Collagenous fibres coming anteriorly and laterally out of disc fibrocartilage provide evidence that sagittal tensile loads are also exerted on discs. Disc dislocations without reduction can only occur when the collagenous fibres coming from the disc also have ruptured. A more critical evaluation of anterior disc dislocations would seem to be called for. PMID- 10474269 TI - [Burkitt lymphoma. Study of 110 patients]. AB - A review of 110 children with Burkitt's lymphoma admitted to the University of Nigeria Teaching Hospital, Enugu during a 7-year period revealed a preponderance of males over females (1.9:1) and a peak incidence at five years. The main sites affected initially by the tumour in order of frequency were the face and abdomen. The spinal column, spleen, and lungs were less affected. It is postulated that an interaction between malarial and Epstein-Barr virus infections and malnutrition are important in the aetiology of the tumour. The most effective drug is cyclophosphamide. A combination of this drug with vincristine and methotrexate is equally effective in inducing remissions, but is better than the single agent in preventing systemic relapse. Follow-up of the patients revealed good survival. PMID- 10474270 TI - [Migration of a retained premolar into the processus muscularis of the mandible]. AB - Migration of an impacted mandibular premolar is a rare finding, whereas dystopia of lower jaw cuspids is described much more often in the literature. We present a patient in whom radiographic studies show the migration of a mandibular premolar to the muscular process over a period of 14 years. Most of the tooth migration along the vertical portion of the mandibula into the processus muscularis took place after the root development had ended. Neither radiological nor clinical examination revealed the stimulus which changed the path and the direction of the impacted tooth. PMID- 10474271 TI - Exogenous estrogens, antiestrogens and lipid metabolism. PMID- 10474272 TI - New aromatase inhibitors for the treatment of advanced breast cancer in postmenopausal women. AB - Inhibition of the enzyme aromatase, the rate limiting step in estrogen production, is an effective endocrine treatment of advanced postmenopausal breast cancer. Recently, several new aromatase inhibitors have been developed to improve efficacy and reduce toxicity compared to the prototype aromatase inhibitor aminoglutethimide. Aromatase inhibitors can be divided into two types. The first are the non-steroidal inhibitors that have a mechanism of action similar to aminoglutethimide. The second are the steroidal inhibitors that function as a false substrate for aromatase. Of the non-steroidal aromatase inhibitors, two drugs, anastrozole and letrozole, have recently been registered for the second line endocrine treatment of advanced postmenopausal breast cancer after failure on tamoxifen. The phase III studies of these drugs indicate at least equal efficacy compared to current second line treatment with aminoglutethimide or megestrol acetate. Their toxicity profile, however, is much more favourable. This makes them the drugs of choice for the second line endocrine treatment of advanced breast cancer in postmenopausal patients, who failed during adjuvant or first line treatment with tamoxifen. Of the steroidal aromatase inhibitors, the orally active drug exemestane is still in phase III clinical study; the registration is expected in 1999. PMID- 10474273 TI - No effect of long-term physical activity on the glycemic control in type 1 diabetes patients: a cross-sectional study. AB - BACKGROUND: The main purpose of the present study was to assess the relation between long-term physical activity and glycemic control in patients with type 1 diabetes mellitus. METHODS: Medical data and blood samples for the determination of glycosylated hemoglobin (glyHb) were collected in 221 consecutive type 1 diabetes patients between 18 and 45 years of age without late complications. A self-report questionnaire was used to determine the degree of physical activity. Correlation coefficients and analysis of variance were used for statistical analyses. RESULTS: No correlation was observed between the different levels of physical activity and glycemic control. Females showed a significantly higher total physical activity index than males (p = 0.004), mostly due to the leisure time activity index. More active patients used a lower amount of insulin (r = 0.20, p = 0.002) than less active patients. CONCLUSIONS: Glycemic control was not found to be associated with long-term physical activity in type 1 diabetes patients. Physical activity did not negatively affect long-term glycemic control. PMID- 10474274 TI - Functional dyspepsia has a good prognosis irrespective of H. pylori status. Long term follow-up of symptoms after anti H. pylori treatment. AB - AIM: A prospective study with anti H. pylori therapy was done in patients with functional dyspepsia. METHODS: Inclusion criterion was the absence of any macroscopic abnormality in oesophagus, stomach, and duodenum, irrespective whether H. pylori was present or not. A questionnaire and a symptom score on a five-point Likert scale were used. Antral biopsy specimens were taken for detection of H. pylori. Treatment consisted of omeprazole 20 mg bid and amoxicilline 500 mg tid during 14 days. Patients were followed during 12 months. At follow-up the questionnaire and the symptom score were used. The main endpoint of the study was clinical remission after 1 year of follow-up. RESULTS: In 1 year 163 patients were included (85 men, 78 women, mean age 47 years, range 21-83 years). H. pylori was present in 91 patients. In the H. pylori positive group 38 patients showed a decrease in IgG antibody titre of more than 50% during follow up of 3-12 months, 26 showed no change or increase following initial decrease. Overall there was no difference in presence or absence of specific complaints at inclusion and final follow-up. The symptom score decreased significantly in all groups (P < 0.001). CONCLUSION: The symptoms of functional dyspepsia improved to a similar extent in both H. pylori positive dyspeptics and the control group. This symptom improvement was irrespective of the change of IgG H. pylori antibodies after therapy. PMID- 10474275 TI - Establishment of reference values for endocrine tests. II: Hyperprolactinemia. AB - BACKGROUND: In patients with hyperprolactinemia, the thyrotropin-releasing hormone (TRH) stimulation test is widely applied to distinguish prolactinoma from other causes of hyperprolactinemia. In the present study, we established reference values for the plasma concentration of prolactin (PRL) and its response to TRH. METHODS: Basal PRL and the PRL response to 400 micrograms TRH i.v. was determined in 50 subjects recruited from the general population, equally distributed according to sex and age between 20 and 69 years. PRL was determined by a fluoroimmunometric assay. Reference values are given as the observed range. RESULTS: Plasma concentrations of PRL were 4.0-25 micrograms/l (median: 10.0 micrograms/l) in women and 0.5-19.0 micrograms/l (median: 8.5 micrograms/l) in men (p = 0.11). The peak PRL concentration after stimulation with TRH was slightly higher in women (median: 51 micrograms/l) than in men (median: 41 micrograms/l; p = 0.04) and was reached at t = 20 min in all subjects. The relative increase in plasma PRL (median: 440%) did not show a statistically significant effect of age or sex. In 12 subjects (24%), the relative increase in plasma PRL was lower than 250%, which has traditionally been considered the minimum cutoff for a normal response. There were no effects of smoking and alcohol, but regular ingestion of liquorice was associated with lower basal (p = 0.03) and lower stimulated (p = 0.05) plasma concentrations of PRL. CONCLUSIONS: The present study provides reference values for basal and TRH-stimulated plasma concentrations of PRL. PMID- 10474276 TI - A patient with diabetes insipidus and periorbital swellings; Erdheim-Chester disease. AB - Erdheim-Chester disease is a rare multisystem disease in which a progressive xanthogranulomatous infiltration of several tissues can be seen. We describe a woman, known to have diabetes insipidus for ten years, with periorbital, retroperitoneal, mediastinal, axillar and inguinal involvement. On histological examination a granulomatous infiltration of fatty tissue and striated muscle was seen, consisting of Touton giant cells, histiocytes with foamy cytoplasm and lymphocytes. Immunohistochemical staining with CD-1a and S-100 was negative and on electron microscopy no Langerhans granules were seen. These findings led to the diagnosis of Erdheim-Chester disease. She had a good response on steroids. Because of some similar clinical features of Langerhans cell histiocytosis and Erdheim-Chester disease, a histiocyte disorder seems the most probable cause. PMID- 10474277 TI - Case report of a patient with an intimal sarcoma of the pulmonary trunk presenting as a pulmonary embolism. AB - A fatal case of an 89-year-old woman with an intimal sarcoma obstructing the pulmonary trunk and an open foramen ovale is presented. Clinical symptoms, physical examination and further evaluation originally raised suspicion of a pulmonary embolism. Recent classification systems, specific radiological and pathological characteristics of sarcomas of the pulmonary trunk are discussed. PMID- 10474278 TI - Tamoxifen and hypertriglyceridemia. AB - A patient developed severe hypertriglyceridemia while on treatment with Tamoxifen after a mastectomy and with a statin for a lipid disorder. The mechanism of this side effect and the therapeutic effect of a fibrate are discussed. PMID- 10474279 TI - Clozapine induced agranulocytosis: hypothesis on immune-mediated pathogenesis and the role of haematopoietic growth factors. PMID- 10474280 TI - The development of a quality of life instrument for use with post-menopausal women with urogenital atrophy in the UK and Sweden. AB - Many post-menopausal women suffer from oestrogen deficiency. This can cause urogenital atrophy which leads to symptoms such as dyspareunia, dysuria, vaginal dryness and urge incontinence. Even though urogenital atrophy is a common condition, little attempt has been made to investigate the impact that the condition has on the quality of life of the women concerned. A quality of life instrument specifically for use with women with urogenital atrophy was developed. The needs-based approach to quality of life was adopted, which states that quality of life is the extent to which an individual is able to satisfy her needs. The development work was undertaken simultaneously in the UK and Sweden. The measure was found to be acceptable and relevant to women in both countries and to have good levels of test-retest reliability (0.92 in the UK and 0.85 in Sweden), internal consistency (alpha coefficients 0.90 in both countries) and construct validity. It is suitable for use in clinical trials and for monitoring the progress of patients in clinical practice. PMID- 10474281 TI - Comparing the psychometric properties of preference-based and nonpreference-based health-related quality of life in coronary heart disease. Canadian Collaborative Cardiac Assessment Group. AB - A cross-sectional survey (n = 878) was conducted to compare the psychometric properties of three preference-based and one nonpreference-based health-related quality of life measures among healthy subjects with and without treatment for dyslipidemia and/or hypertension and patients with coronary heart disease (CHD). All measures were stable over a 3 to 6 week period. Compared to the Time Trade off (TTO) and the Standard Gamble (SG), the Rating Scale (RS) correlated with the SF-36 Health Survey most highly. In contrast to the SF-36 General Health Perception (GHP), the SF-36 Physical Component scale and the RS, the TTO and SG were less able to discriminate CHD patients with various levels of physical disability. Only the SF-36 GHP subscale and the RS were able to differentiate healthy participants from participants receiving dyslipidemia and/or hypertension treatment. Neither the SF-36 Physical or Mental Component scales were able to discriminate these two groups. Overall, these results suggest that unlike the RS, the TTO and the SG, as administered in this study, may not be sufficiently sensitive to measure the impact of primary cardiovascular disease prevention strategies on the health-related quality of life of the participants. PMID- 10474282 TI - Quality of life of women with systemic lupus erythematosus or rheumatoid arthritis: domains of importance and dissatisfaction. AB - The aim of this study was to increase understanding of the domains of life that are important to Swedish women with chronic rheumatic disease and to describe their dissatisfaction with these domains. Fifty women with Systemic Lupus Erythematosus (SLE) and 50 with Rheumatoid Arthritis (RA) were interviewed by telephone regarding their quality of life using five open-ended questions. Content analysis of their responses revealed nine categories: Health/wellness, Family/friends, Work, House/home/living environment, Social support/functional services, Hobbies/cultural activities, Sufficient income, Independence, Integrity/identity. Areas of importance correspond closely to those areas with which they were most dissatisfied. Although the domains identified in both groups were the same, SLE patients expressed more dissatisfaction with their perceived control over their bodies and understanding about their condition on the part of physicians and people in general. RA patients were more dissatisfied with areas of their lives that threatened their independence. Health care providers should be aware of these areas of dissatisfaction so that they can plan strategies to maximize patient quality of life. PMID- 10474283 TI - Olanzapine versus haloperidol in the treatment of schizophrenia and other psychotic disorders: quality of life and clinical outcomes of a randomized clinical trial. AB - BACKGROUND: Little information is available on the impact of the atypical antipsychotic olanzapine on quality of life (QOL). A 6-week, double-blind randomized multicenter trial, with a long-term extension, was conducted to evaluate the clinical efficacy and QOL of olanzapine and haloperidol in treating schizophrenia and other psychotic disorders. METHODS: A total of 828 outpatients provided QOL data. Study patients were aged greater than 18 years with a DSM-III R diagnosis of schizophrenia, schizophreniform disorder, or schizoaffective disorder and baseline BPRS (items scored on 0-6 scale) total scores, > or = 18 were randomized to 6 weeks of treatment with olanzapine 5 to 20 mg/day or haloperidol 5 to 20 mg/day. Patients entered a 46-week double-blind extension if they demonstrated minimal clinical response and were tolerant to study medication. The Quality of Life Scale (QLS) and SF-36 Health Survey were used to evaluate QOL. RESULTS: During the 6-week acute phase, olanzapine treatment significantly improved BPRS total (p = 0.004), PANSS total scores (p = 0.043), QLS total (p = 0.005), intrapsychic foundations (p < 0.001) and interpersonal relations scores (p = 0.036), and SF-36 mental component summary scores (p < 0.001) compared with haloperidol. During the extension phase, olanzapine treatment significantly improved PANSS negative scores (p = 0.035) and improved QLS total (p = 0.001), intrapsychic foundations (p < 0.001), and instrumental role category scores (p = 0.015) versus haloperidol treatment. Significantly more haloperidol patients discontinued treatment due to adverse events during the acute and extension phases (p = 0.041 and p = 0.014, respectively). Changes in QLS total and MCS scores were associated with changes in clinical symptoms, depression scores and extrapyramidal symptoms. CONCLUSIONS: Olanzapine was more effective than haloperidol in reducing severity of psychopathology and in improving QOL in patients with schizophrenia and other psychotic disorders. The QOL benefits of olanzapine, although modest, may be important for long-term treatment. PMID- 10474284 TI - Coping and quality of life in patients with psoriasis. AB - The aim of the present study was to investigate the relationship between coping dimensions and overall quality of life, disability and health status in patients' with psoriasis. Psoriasis is one of several chronic diseases which requires self management in order to ensure an enhanced quality of life. The sample comprised 334 patients who were treated consecutively at three dermatology departments in eastern Norway. A total number of 273 patients completed the questionnaire, yielding a response rate of 82% (20% in-patients and 80% out-patients). The following questionnaires were used: The Jalowiec Coping Scale, the Psoriasis Disability Index, the Quality of Life Scale, and the SF-36. Results showed that patients who used combined emotive coping strategies reported more disability, poorer mental health and worse overall quality of life. Furthermore, patients who more frequently used normalising/optimistic coping reported higher levels of mental health. However, the variance explained by coping effort was low to moderate. Coping explained the variance in mental health and overall quality of life to a greater extent than that in physical health. Knowledge about the relationships between coping and quality of life dimensions is important with regard to the establishment and implementation of appropriate psychosocial interventions for patients with psoriasis. PMID- 10474285 TI - Adapting the Jalowiec Coping Scale in Norwegian adult psoriasis patients. AB - The aim of the present study was to adapt the Jalowiec Coping Scale (JCS) to accommodate adult patients with psoriasis. The sample comprised 334 patients who were treated consecutively at three dermatology departments in the eastern Norway. A total number of 273 hospitalised patients (20%) and out-patients (80%) completed the questionnaire, yielding a response rate of 82%. The study assessed the reliability and the face, content and construct validity of the Norwegian version of the JCS. In addition, researchers investigated the most frequently used/effective coping strategies, the relationships between demographic/clinical variables, self-reported physical symptoms and the use of coping strategies. The results (correlational coefficients and interitem alpha s) indicated that there was an overlap in substantive content among the original JCS subscales, due either to measurement error (bias or response style) and/or because the patients in the present study were in a demanding situation in relation to their disease, which may have activated a variety of coping strategies. A factor analysis resulted in a three-factor solution (confrontive problem-solving, normalising/optimistic and combined emotive) with satisfactory internal consistency. This factor solution comprised 31 items with an explained variance of 37% of the total pool of items. The most frequently used and effective coping strategies could be labelled as emotion-focused (optimistic/maintain control). Significant correlations were found between age, hospital setting, self-reported physical symptoms and different coping subscales. However, further studies are needed to assess the validity and reliability of the JCS among different population groups in Norway. PMID- 10474286 TI - Distinguishing between quality of life and health status in quality of life research: a meta-analysis. AB - Despite the increasing acceptance of quality of life (QOL) as a critical endpoint in medical research, there is little consensus regarding the definition of this construct or how it differs from perceived health status. The objective of this analysis was to understand how patients make determinations of QOL and whether QOL can be differentiated from health status. We conducted a meta-analysis of the relationships among two constructs (QOL and perceived health status) and three functioning domains (mental, physical, and social functioning) in 12 chronic disease studies. Instruments used in these studies included the RAND-36, MOS SF 20, EORTC QLQ-30, MILQ and MQOL-HIV. A single, synthesized correlation matrix combining the data from all 12 studies was estimated by generalized least squares. The synthesized matrix was then used to estimate structural equation models. The meta-analysis results indicate that, from the perspective of patients, QOL and health status are distinct constructs. When rating QOL, patients give greater emphasis to mental health than to physical functioning. This pattern is reversed for appraisals of health status, for which physical functioning is more important than mental health. Social functioning did not have a major impact on either construct. We conclude that quality of life and health status are distinct constructs, and that the two terms should not be used interchangeably. Many prominent health status instruments, including utility based questionnaires and health perception indexes, may be inappropriate for measuring QOL. Evaluations of the effectiveness of medical treatment may differ depending on whether QOL or health status is the study outcome. PMID- 10474288 TI - [Biological effect of exposure to butadiene]. AB - People can be exposed to 1,3-butadiene in work place (rubber industry) as well as in natural environment (car exhausts fumes, cigarette smoke). Butadien on its own is not genotoxic, but is metabolized to mutagenic and carcinogenic epoxydes, 1,2,3,4-diepoxybutane and 3,4-epoxybutene in the organism of mammals and human. 1,3-butadiene has been shown to be a potent carcinogen in animals and human. Laboratory investigations showed also toxic and mutagenic abilities of butadiene and its metabolities. Interspecies differences in sensitivity to butadiene are caused by differences in metabolic transformations of butadiene in different species. PMID- 10474287 TI - Quality of life in low-income patients with metastatic prostate cancer: divergent and convergent validity of three instruments. AB - Few studies have evaluated Quality Of Life (QOL) among low-income patients with cancer. Information is needed about the feasibility and psychometric characteristics of QOL instruments in these populations. The purpose of this study was to examine the convergent and discriminant relationships between scales of three QOL instruments: European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC), Functional Assessment of Cancer Therapy--General (FACT), and Quality of Life Index (QLI). Participants included 110 men with metastatic prostate cancer of whom 94% were low income and 62% were African-American. Interviewers administered the questionnaires. Cronbach alpha internal consistency reliabilities were 0.57 to 0.90 for the EORTC, 0.65 to 0.86 for the FACT, and 0.63 for the QLI. Convergent validity was supported for the EORTC and FACT scales measuring emotional, physical, and role/functional dimensions (r = 0.54 to 0.72), but not on scales measuring social function (r = 0.12). Divergent validity was supported between dissimilar scales (r = 0.14 to 0.38). Analysis with receiver operating characteristics curves provided empirical support for the EORTC and FACT as multidimensional measures. These findings suggest that, even in busy clinical settings with low literacy patients, interviewer-administered EORTC and FACT QOL instruments can provide valid and reliable information. PMID- 10474289 TI - [Content of nitrates and nitrites in market vegetables in Wroclaw in the years 1996-1997]. AB - The aim of analytical investigation included in this paper was the determination the nitrate and nitrite contents in 8 common vegetables: cabbage, carrot, parsley, potato, garden beet, cucumber, lettuce and radish in relation to the acceptable levels of nitrates. The samples of vegetables were taken from detail shops in Wroclaw in the years 1996-1997. A total of 151 samples of vegetables were analyzed. The nitrate and nitrite level was determined collorimetricaly with sulfanilic acid after previous reduction of nitrates to nitrites by means of cadmium column. The acceptable values of nitrates were exceeded in 82% of the samples of radish and in 65% samples of lettuce. The smallest amount of samples with excess of the acceptable nitrate levels were found in potato, carrot and cucumber. It was found, that the nitrate content in the greenhouse vegetables: lettuce, cucumber and radish was greater than in the field vegetables. The potato, cabbage and carrot investigated after winter storage comprised lower content of nitrates than in freshly harvested. Nitrite accumulation in investigated vegetables was low, but the highest concentrations were found in lettuce and radish form greenhouse. PMID- 10474290 TI - [Levels of organochlorine insecticides in citrus fruits in 1996-1997]. AB - The monitoring of organochlorine insecticides (HCB, HCH, and DDT) in citrus fruits taken from the market was carried out in 9 regions in Poland in the years 1996-1997. The concentrations of the insecticides were measured in the edible part and in the peel separately. The total number of 331 samples of oranges, lemons, grapefruits and mandarins were subjected residue analysis. The mean concentrations of the organochlorine compounds in the edible parts amounted 0.0002 mg/kg for HCB, and ranged from 0.0081 to 0.0247 mg/kg for sigma HCH and from 0.0041 to 0.022 for DDT depending on the product. The highest contrentrations together with the highest violation rate (9.5%) of the Polish Maximum Residue Limit was found in case of DDT. The mean concentrations of the insecticides in the citrus peels were not markedly higher than those in the edible parts. The results obtained in this monitoring did not differ from the results reported by the other authors. PMID- 10474291 TI - [Residues of tris(4-chlorophenyl)methane and tris(chlorophylyl)methanol in fish from the Gulf of Gdansk]. AB - Tris(4-chlorophenyl)methane (TCPM-H) and tris(4-chlorophenyl)methanol (TCPM-Oh) were identified and quantified in fish caught in the Gulf of Gdansk, Baltic Sea. The method of measurement was capillary gas chromatography and low resolution mass spectrometry (HRGC/LRMS) after a nondestructive extraction and clean-up of the samples using a wide-bore glass tubes and dialysis with a semipermeable polyethylene membrane and further fractionations of the extract on Florisil column. TCPM-H and TCPM-OH were detected in fish in concentration from 0.08 to 1.6 and from < 0.02 to 0.4 ng/g wet weight, respectively. PMID- 10474292 TI - [Organochlorine pesticides and polychlorinated biphenyls in ambient air in the city of Gdansk]. AB - The concentrations of organochlorine pesticides such as DDTs, HCHs, CHLs, HCB and polychlorinated biphenyls (PCBs) were determined in ambient are samples collected in city of Gdansk in 1991-1992 to understand concentrations, sources and seasonal distribution. Polyurethane foam plugs were used as adsorbents for collection of persitent organochlorines in ambient air samples. Identification and quantification of organochlorines were carried out using a capillary column gas chromatography and ECD (63Ni) detection. HCB, alpha-HCH, gamma-HCH, p,p'-DDT, o,p'-DDT, p,p'-DDE, o,p'-DDE, p,p'-DDD, o,p'-DDD and PCBs were detected in all samples, while the constituents of technical chlordan were absent in concentration above the detection limit of the method (trans-chlordane < 14 pg/m3, cis-chlordane < 8.1 pg/m3 and trans-nonachlor < 7.4 pg/m3). The concentrations of HCB and PCBs were depended on the air temperatures, hence their main source can be related to a process of degasing from the soil. PMID- 10474293 TI - [Levels of total mercury in scalp hair of children and youth from the selected rural area of the Lublin district]. AB - The total mercury content in the hair of 195 children and youth aged 4-15 years was determined using mercury analyzer AMA 254 (ALTEC-Czech Republic). The examined individuals lived on the rural area of south-east Lublin district. The analysis of the results did not revealed essential differences in the mean concentrations of mercury according to sex, age, colour of hair and dental amalgam fillings. There was found that average hair mercury level was higher in the group often consuming fish, mushrooms, pluck and giblets compared with the group prefering another kind of diet. The same conclusion was drawn in relation to the exposed and non-exposed to cigarette smoke groups. PMID- 10474294 TI - [The review of the European Union documents relating to contamination of aflatoxins in food]. AB - In the following review, Polish and European Union legislation, concerning maximum level of aflatoxins in foodstuffs, was reported. In this moment, no specific requirements exist but according to Food and Nutrition Law all kind of food ought to be free from aflatoxins. Practically, maximum aflatoxins level should be below the detection limit of official analytical method (thin layer chromatography)--5 micrograms/kg and 0.05 microgram/l for milk. Commission Directive 98/53/EC laying down the sampling methods and the methods of analysis for aflatoxins in foodstuffs and Commission Regulation No 152/98 setting maximum levels for aflatoxins in foodstuffs were presented. Also, Ministry of Health Draft Regulation, concerning maximum levels of contamination in food is prepared in accordance with EU regulation, was reported. PMID- 10474295 TI - [Solanine and chaconine: occurrence, properties, methods for determination]. AB - Glycoalkaloids are naturally occurring toxicants in plants that are members of the Solanaceae family. In this paper occurrence of glycoalkaloids, especially solanine and chaconine in potatoes and tomatoes, were reviewed. Basing on literature, toxicological properties and methods of determination were reported. Attention was paid to common content of glycoalkaloids in potatoes and tomatoes and their commercial products. Solanine and chaconine are usually present at low levels in large majority of current commercial varieties but they can accumulate to high levels in greened, stored, damaged potatoes. High concentration may cause acute poisoning, including gastro-intestinal and neurological disturbances, in man. The upper limit, recognized as a safe (non-toxic), was presented. According to WHO normal levels in potatoes 20-100 mg per kg of potatoes is not of toxicological concern. PMID- 10474296 TI - [Determination of vitamin C in selected fruit and vegetable products]. AB - A simple method was described to determine vitamin C as L-ascorbic acid (after reduction of dehydroascorbic acid by means of dithiothreitol) in fruit juices, fruit and vegetable-fruit nectars. Ascorbic acid was analyzed by RP-HPLC technique with UV detection (254 nm). The average recovery of ascorbic acid was 92-103% and limits of identification and detection were 0.003 and 0.009 mg/ml of products respectively. PMID- 10474297 TI - [Nutritional importance of oligosaccharides]. AB - Oligosaccharides are widely distributed in higher plants, especially leguminous seeds. This review described the structure of galactooligosaccharides, fructooligosaccharides and raffinose-type oligosacchariddes. Flatulance--causes, relation to diet and composition of intenstinal gas are discussed. Emphasis is placed upon the fact that ingestion of oligosaccharides increases the bifidobacteria population in the colon, which in turn contributes to human health in many ways. PMID- 10474298 TI - [Recommendation for validation and routine monitoring of sterilization processes with ethylene oxide for medical devices]. AB - The European Medical Device Directives specifically address sterilization issues in a number of instances. The European Standards for sterilization of medical devices, especially EN 550, EN 554, EN 556 regulate the manufacture, installation and operation of sterilizers as well as the validation of sterilization processes, on using ethylene oxide (EN 550) or moist heat (EN 554) for sterilization. This recommendation is intended as a source of information for conducting validation according to EN 550 and concomitantly for ensuring that the medical devices reprocessed (cleaned, disinfected, packed, sterilized, stored) in the hospital setting or in other healthcare establishments are endowed with the same level of safety with respect to sterility as that of industrially produced and marketed sterile medical devices. PMID- 10474299 TI - [Children nutrition and food preparation conditions in orphanages]. AB - The study was carried out in all orphanages listed in sanitary-epidemiological stations. The main purpose of the study was to assess children nutrition and sanitary-hygienic conditions of meal's preparation in orphanages. Inspectors of Children's and Youth's Hygiene Departments of the sanitary-epidemiological stations assessed, by means of a uniformed questionnaire, the sanitary-hygienic conditions of kitchens and dinning-rooms and quality of diet served in orphanages. From total 327 children's homes (with 16471 inmates aged from 3 days up to 25 years) included in the study, 93% were run by State, 6% by the Church and 1% by Foundations. Results of the study showed that in majority of orphanages the sanitary conditions of meal preparation and consumption were satisfactory. Kitchens with proper equipment were recorded in 301 orphanages (92%), with good ventilation in 316 orphanages (97%) and with sufficient lighting in 304 orphanages (93%). Dietary assessment of children showed that in some orphanages inmates have eaten too few meals a day. In contrary it must be emphasized that in 241 orphanages (73%) the meals were served five times a day. It leads to assumption that in remaining 27% such a way of meals giving should be possible to achieve. PMID- 10474300 TI - Role models of the female persuasion. PMID- 10474301 TI - Women in medicine: still a long way to go, baby. AB - No doubt about it, women have come a long way in the medical profession. The average female enrollment in our nation's medical schools is 42% and growing. Women physicians are entering more non-traditional fields and are running successful practices. They're working to change attitudes and overcome obstacles that have traditionally barred them from thriving in such a demanding field. They're getting creative with options that allow them to pursue their dream while raising a family. But the bars are still up on some windows, namely the ones that peek into the halls of organized medicine. Professional organizations and medical schools are still largely run by men. The Tennessee Medical Association recognizes the discrepancy and is working to make some changes. TMA's Executive Assistant for Membership Services Phyllis Franklin says leadership is rethinking the entire structure of the Association in the light of changing demographics. "We're looking at younger physicians, minorities, female physicians, academicians, and students; we want to get them involved in organized medicine," she says. The leadership structure is one area that is slowly changing. Franklin admits while the hierarchy has been male-dominated, the Association can now claim its first female vice president, Dr. Phyllis Miller of Chattanooga. "We don't have a lot of women leaders, but it's getting better, and there are a lot more opportunities in the committee structure." TMA officials say surveys are forthcoming that will help them gauge the needs and wants of its female membership and hopefully increase further that 11% number, up from 8.9% two years ago. In the meantime, Franklin encourages women physicians to make their wishes known. "Instead of not joining the TMA because of perceived barriers or stumbling blocks, we would certainly prefer that they help us identify obstacles and give us the opportunity to do what we can to make membership more appealing," she says. "This Association is truly for all physicians." PMID- 10474302 TI - Business and medicine. PMID- 10474303 TI - Planning physician supply: too hot, too cold, or just right? PMID- 10474304 TI - Loss prevention case of the month. Learn from and respond to the record. PMID- 10474305 TI - Accidental veterinary antibiotic injection into a farm worker. AB - A 29-year-old white male farm worker accidentally injected tilmicosin, a bovine antibiotic, into his finger. He developed temporary pulmonary, gastrointestinal, and neuromuscular symptomatology and a more persistent subjective asthenia. PMID- 10474306 TI - Hypoparathyroidism, intracranial calcification, and seizures 61 years after thyroid surgery. AB - Though hypocalcemic symptoms from hypoparathyroidism following thyroidectomy most often occur in the immediate postoperative period, hypoparathyroidism can present itself many years after the thyroid surgery with nonspecific symptoms. We present herein the case of a 74-year-old woman with previously undiagnosed hypoparathyroidism who had tonic-clonic seizures and intracranial calcification 61 years after her thyroidectomy. This case is unusual because of the long latency between thyroidectomy and clinical presentation. PMID- 10474307 TI - A case of alcohol withdrawal. PMID- 10474308 TI - Medicare rural hospital flexibility program. PMID- 10474309 TI - Diagnosis and management of dementia in primary care at an early stage: the need for a new concept and an adapted procedure. AB - Diagnosis of dementia in primary care is both difficult and important. The recommendations by several authors to improve the diagnosis of dementia by general practitioners are important, but insufficient. It is argued that perhaps the disease concept in itself is a cause of confusion for clinicians. Primary care physicians need an adapted procedure, gradually leading to the final diagnosis of dementia. It has to be a stepwise labelling strategy, using global descriptions and non-disease specific labels in the beginning, ending up with well-defined disease criteria. In this process, there is circularity: previous diagnoses have to be kept in mind because symptoms and signs may gradually change during the progression of the disease, leading to reconsideration of previous deleted options. To frame this properly, the primary care physician needs to adopt a broad "frail elderly" geriatric concept. Implementation of this concept not only helps the diagnostic process, but also stimulates the care for dementing patients and their caregivers. Relevant arguments for early diagnostic involvement of primary care physicians can be put forward on condition that a new concept, adapted procedures and adapted instruments are used. PMID- 10474310 TI - The role of science in medicine. AB - A suitable demarcation between pure science and applied research can be drawn in terms of their goals. This distinction of goals has methodological and cultural consequences. If the demarcation is accepted, what does the connection between the two enterprises look like? What is the role of science in medical practice? The Baconian answer to this question is discussed and criticised as too linear. A second answer may be that pure science has no part at all in medicine. This too can be criticised as too simplistic. A third answer is suggested in which pure science plays the role of being an instrument for interpreting observations and problems, and of being a source of inspiration for technological research programs. PMID- 10474311 TI - Sex biases in subject selection: a survey of articles published in American medical journals. AB - This study discusses the results of a survey of 1,800 articles published in American medical journals from 1985-1996. The study finds 9% of these articles reported research that uses only male subjects to examine medical conditions that affect both sexes; the ratio of research on female to male conditions among these articles was greater than 5:1; but 76.5% of the articles reported research that includes both male and female subjects. The study also discusses evidence that sex biases against women (and men) are decreasing. This study also offers some possible psychological, institutional, medical, and economic explanations of the sex biases in medical research published in American journals, and discusses some policy implications of sex biases in medical research. The study concludes by urging others to conduct more empirical research on sex biases in medical research. PMID- 10474312 TI - The dream of consensus: finding common ground in a bioethical context. AB - Consensus is the holy grail of bioethics, the lynch pin of the assumption that well informed, well intentioned people may reach generally acceptable positions on ethically contentious issues. It has been especially important in bioethics, where advancing technology has assured an increasing field of complex medical dilemmas. This paper results on the use of a multicriterion decision making system (MCDM) analyzing group process in an attempt to better define hospital policy. In a pilot program at The Hospital for Sick Children, Toronto, a series of small scale focus groups was constituted to examine criteria defining organ transplant eligibility. Criteria were organized hierarchically using the Analytic Hierarchy Process, an MCDM approach, and the resulting data was analyzed using Expert Choice 9.0, software designed to facilitate AHP analysis. Qualitative and quantitative analysis map barriers to practical consensus in a way not previously possible. PMID- 10474314 TI - Assisted suicide, suffering and the meaning of a life. AB - The ethical problems surrounding voluntary assisted suicide remain formidable, and are unlikely to be resolved in pluralist societies. An examination of historical attitudes to suicide suggests that modernity has inherited a formidable complex of religious and moral attitudes to suicide, whether assisted or not. Advocates usually invoke the ending of intolerable suffering as one justification for euthanasia of this kind. This does not provide an adequate justification by itself, because there are (at least theoretically) methods which would relieve suffering without causing the physical death of the suffering person. Carried to extremes, these methods would finish the life worth living, but leave a being which was technically alive. Such acts, however, would provide no moral escape, since they would create beings without meaning. Arguments seeking to justify ending the lives of others need some grounding in concepts of the meaning of a life. The euthanasia discourse therefore needs to take at least some account of the meaning we construct for our lives and the lives of others. PMID- 10474313 TI - The (gendered) construction of diagnosis interpretation of medical signs in women patients. AB - Medicine maintains a distinction between the medical symptom--the patient's "subjective" experience and expression, and the privileged medical sign--the "objective" findings observable by the doctor. Although the distinction is not consistently applied, it becomes clearly visible in the "undefined," medically unexplained disorders of women patients. Potential impacts of genderized interaction on the interpretation of medical signs are addressed by re-reading the diagnostic process as a matter of social construction, where diagnosis results from human interpretation within a sociopolitical context. The discussion is illustrated by a case story and empirical evidence of the gendering in the doctor-patient relationship. The theoretical analysis is supported by semiotic perspectives of bodily signs, feminist theory on experience, and Foucault's ideas about medical perception and gaze, and concludes that a medical diagnosis is seldom a biological fact, but the outcome of a process where biological, cultural and social elements are interwoven. Further deconstruction of the chain of signs from a feminist perspective, assigning validity to the voice of the woman patient, might broaden the understanding of women's health, illness and disease. PMID- 10474318 TI - [Egocentric inference of object positions during reading and listening]. AB - Subjects read narratives describing several objects surrounding a protagonist. In taking the protagonist's perspective, subjects constructed egocentric mental models of the described object configurations. During the test phase, the mental model had to be updated according to reorientations of the protagonist. We examined if objects are localized immediately within the egocentric reference frame of the mental model, even if they are not linguistically localized relative to the protagonist, that is, if egocentric object positions have to be inferred. In two experiments, narratives and test items were presented either visually or aurally. In a third experiment, narratives were presented visually but test items were presented aurally. We found that recipients tended to localize every object immediately within the egocentric reference frame. But since reading interferes with updating of the mental model, subjects did not localize every object egocentrically if visual presentation was used throughout. PMID- 10474320 TI - [The crossover effect of the propositional text representation and the mental model:the role of individual differences in spatial imagery ability]. AB - The crossover effect (Mani & Johnson-Laird, 1982) predicts that a mental model is more likely to be available after reading a determinate description of a spatial scene than after reading an indeterminate description whereas the reverse should apply to the propositional text representation. Although attempts to replicate this pattern were less than convincing, the crossover effect has often been stated as evidence justifying the differentiation of propositional representations and mental models. In an experiment with 44 adult, German speaking participants, it was investigated whether the crossover effect is moderated by individual differences in spatial imagery ability. Participants read 6 determinate and 6 indeterminate descriptions of schematic, non-schematic, or schema-incompatible spatial scenes. Recognition data showed a crossover effect only in participants with lower ability. In the high-ability group the propositional text representation was less available than in the low-ability group, independent of whether or not a mental model could be constructed. The results explain why earlier replications might have failed. Consequences for current mental model theories of text comprehension are discussed. PMID- 10474321 TI - [Strategic focusing of attention during narrative reading]. AB - In 2 experiments based on the constructionist view of text comprehension, we investigated whether readers are able to strategically use, in a problem solving matter, spatial and temporal information that varies in its relevance to the goals of the protagonists. In order to accomplish this, we varied the specificity of the protagonists' goal descriptions. Both experiments consisted of a reading section, a test, and a questionnaire, and differed only in respect to when the questionnaire was administered. The results of both experiments showed that readers were able to use spatial and temporal information of narrative texts in a strategic matter, and they even did so without explicit instruction. This focus of attention, however, was not uniformly reflected in the reading times. Memory data showed a clear disadvantage for temporal information as compared to spatial information. This was the case even though both types of information had been equally well identified in the questionnaire as crucial to the problem solving process of the protagonist. PMID- 10474322 TI - [Situation models in text comprehension: will emotionally relieving information be automatically activated?]. AB - It was tested whether the "situation model" framework can be applied to research on coping processes. Therefore, subjects (N = 80) were presented with short episodes (formulated in a self-referent manner) about everyday situations which potentially ended in a negative way (e.g., failures in achievement situations; losses etc.). The first half of each episode contained a critical sentence with emotionally relieving information. Given a negative ending, this information should be automatically activated due to its relieving effect. A two-factorial design was used. First, a phrase from the critical sentence was presented for recognition either after a negative ending, a positive ending, or before the ending. Second, with minor changes a control sentence (with an additionally distressing character) was constructed for each potentially relieving sentence. As hypothesized, an interaction emerged: Given a negative ending, the error rate was significantly lower for relieving information than for the control version, whereas there was no difference if the test phrase was presented before the end or after a positive end. PMID- 10474324 TI - [Influence of the type of visualization on the construction of mental models during picture and text comprehension]. AB - The following article describes an integrative model of text and picture comprehension. Text and picture comprehension is considered as a task oriented process of constructing descriptive and depictive mental representations by selection and organisation of information, parsing of symbols, mapping of analog structures as well as mental model construction and model inspection. Based on this theoretical model an experiment was conducted in which subjects had to acquire knowledge about a complex learning content (time and date differences on the earth) through self directed learning with a hypertext and different, but informationally equivalent pictures. The study aimed to investigate under which conditions learners prefer which kind of information and whether the kind of visualisation has an effect on the mental model structure created during the comprehension of texts and pictures. The results indicate that simple pictures induce more superficial processing, in which picture comprehension replaces text comprehension to some extent and vice versa. Demanding pictures, on the contrary, induce more intensive processing in which text comprehension and picture comprehension stimulate each other. Furthermore, the results suggest that the surface structure of pictures affects the structure of mental models. Accordingly, the presentation of a visualisation which is not task adequate may interfere with the required mental model construction. In designing texts combined with pictures the form of visualisation deserves therefore special attention. PMID- 10474327 TI - The problem with Cytotec is that it has not been approved for obstetrical use. PMID- 10474328 TI - Interview with Margaret Nofziger. Interview by Stephanie Bonser. PMID- 10474329 TI - Randomized controlled trials as authoritative knowledge. Keeping an ally from becoming a threat to North American midwifery practice. PMID- 10474330 TI - Rho(D) immune globulin: pros, cons, indications, and alternatives. PMID- 10474332 TI - Rhiannon's water birth. A homebirth in military housing. PMID- 10474331 TI - One mother's experience with Rhesus isoimmunization. An interview with Marlene Gallichan. Interview by Jane Bernstein. PMID- 10474333 TI - Women of the thirteenth moon. The menopause experience. Part I. PMID- 10474334 TI - The Gaskin maneuver article. PMID- 10474335 TI - Transformational birth opportunities: making way for more spiritual births. PMID- 10474336 TI - Every birth is sacred: helping each other through the pain of cesarean section. PMID- 10474337 TI - Women of the thirteenth moon: the menopause experience. Part II. PMID- 10474338 TI - The ties that bind medical intervention and maternal mortality: Mary's story. PMID- 10474339 TI - Mother's milk soap. The most precious soap in the world! PMID- 10474340 TI - Midwifery and the gift of healing touch. PMID- 10474341 TI - Genetic screening. PMID- 10474342 TI - Cover your aura: life with malpractice insurance. PMID- 10474343 TI - Healing tales: a view from another small island. PMID- 10474344 TI - Getting from here to there. PMID- 10474345 TI - Using homoeopathy to offer supportive cancer care, in a National Health Service outpatient setting. AB - This article explores the role of homoeopathy in supporting the patient with cancer. The author describes the homoeopathic approach and clinical research into its use in the cancer setting. A case history illustrates the practical application of the homoeopathic method and its success in managing symptoms of fatigue and anxiety. The author discusses potential problems using the approach. A wider discussion follows looking at why complementary therapies offer an important means to support the patient through different phases of the cancer diagnosis. There is an emphasis on the need for research to clarify whether conventional and complementary therapies can impact on survival and support wellbeing. PMID- 10474346 TI - 'Just the ticket': case studies, reflections and clinical supervision (Part III). AB - An earlier paper in this series of three has detailed the background development and organization of a pilot complementary therapy service. The setting was a neuroscience ward with existing staff carrying out the treatments. Outcome measures including physiological indices and patient feedback were reported on in Part II (Dryden et al. 1999). This paper will now focus on both the records of the practitioner's reflections and the recurring themes of their monthly clinical supervision sessions. Subgroups of four patients with Guillain-Barre Syndrome (GBS) are discussed in detail, as they received the most number of treatments between them. PMID- 10474347 TI - Using complementary therapies within nursing: some ethical and legal considerations. AB - Increasing numbers of health professionals are incorporating complementary and alternative (CAM) therapies into their work. Nurses, in particular, are embracing CAM techniques, which may fit well within their philosophy of care for their patients and which may, instinctively, be more holistic than the approach of their medical counterparts. Patients, who may be distillusioned with conventional approaches to treatment, appear to express high levels of satisfaction in receiving these therapies and nurses clearly enjoy giving them. Beyond patient preference for CAM therapies though, there is still a paucity of clinical evidence to support the wider integration of CAM into the NHS. This article focuses on some of the ethical and legal considerations raised by nurses who employ, or are considering employing, CAM techniques as part of their NHS work, looking, in particular, at the need to ensure patient safety whilst at the same time protecting nurses from criticism or censure. PMID- 10474348 TI - Auditing complementary therapies in palliative care: the experience of the day care massage service at Mount Edgcumbe Hospice. AB - The bulk of this paper comprises an audit of the massage service offered to day care clients at Mount Edgcumbe Hospice. Having had no experience of audit it has been a learning process within which shortcomings of my techniques have become evident. However, during the course of the project, I became enthusiastic about the potential for change, progress and increased validation enabled by the audit process. I realised the significance and importance of audit as an evaluatory tool in relation to complementary and alternative medicine. Still in the earlier stages of acceptance by more orthodox medicine, these therapies require audit, as much as they require quality scientific research. 'Complementary and alternative medical practitioners, teachers and researchers need to understand the advantages of more systematic audit and rigorous research' (Foundation for Integrated Medicine) The objectives of this audit are: To review the performance of the massage in day-care after 2 years of activity. To obtain first hand feedback from clients, staff and carers with a view to introducing improvements. To devise a system of measuring the performance of the day-care service both for comparison internally and with other external hospices. To benefit from the discipline which the audit process itself engenders in complementary medical practice. To lend credibility to the massage service and other complementary therapies in the context of more orthodox palliative care. To arrive at a method of measuring massage as a complementary therapy. To improve communication between the different areas of the hospice. I would encourage other complementary therapists to follow suit and start auditing to help validate the importance of these therapies in today's more holistic approach to health. PMID- 10474349 TI - Technical approach to care. PMID- 10474350 TI - Try a little tenderness. PMID- 10474351 TI - Major incident planning particularly those including chemicals. AB - A chemical incident can happen anytime, anywhere and is often complex in nature. Henrietta Wheeler advises on the necessary precautions staff must take to avoid the risk of poisoning and contamination to both staff and casualties. PMID- 10474352 TI - Running a seminar. AB - A seminar may be less complicated to arrange than a study day but there are still things that can go wrong. Rosemary Cook outlines what is essential to a successful presentation. PMID- 10474353 TI - Assessing deliberate self harm: a team approach. PMID- 10474354 TI - Health promotion in relation to domestic violence. PMID- 10474355 TI - Ending violence in A&E. PMID- 10474356 TI - Besieged. AB - Una Bell is service director of A&E at Whipps Cross Hospital in London. On a Monday afternoon in June last year, she invited an angry woman into her office who wanted to make a complaint to a manager. What follows is her personal account of being held hostage and how the police and the courts responded to her harrowing ordeal. PMID- 10474357 TI - The UKCC's work on nurse practitioners and clinical nurse specialists. PMID- 10474358 TI - A guide to defibrillation. PMID- 10474359 TI - Critical incident stress management strategies. PMID- 10474360 TI - The nurse as team leader during advanced life support in A&E. PMID- 10474361 TI - Dynamic process of triage. PMID- 10474362 TI - ER. You watch it ... we live it! PMID- 10474363 TI - ENP education: a United States perspective. PMID- 10474364 TI - Abdominal examination and assessment in A&E. PMID- 10474365 TI - 'Keep asking things' capturing clinical ideas. PMID- 10474366 TI - Adolescence: the forgotten factor. PMID- 10474367 TI - Developing integrated emergency care. PMID- 10474368 TI - Avalon or Atlantis? Emergency nursing at Glastonbury 1998. PMID- 10474369 TI - Caring for the patient with Alzheimer's Disease. AB - Treating the patient with Alzheimer's disease requires compassion and creativity. Polly Zimmermann and Anna Ortigara outline some inventive care strategies. PMID- 10474370 TI - How medical negligence is established. PMID- 10474371 TI - Treating the child with minor scalp wounds. PMID- 10474372 TI - Communication and trauma management. PMID- 10474373 TI - St. Augustine's, a South African trauma unit. PMID- 10474374 TI - Managing actual and potential hostage taking incidents. PMID- 10474375 TI - Systematic eye examination in A&E. PMID- 10474376 TI - Caring for children. PMID- 10474377 TI - Principles of universal precautions. PMID- 10474378 TI - Nursing management of minor burn injuries. PMID- 10474379 TI - Pregnancy and punishment. PMID- 10474380 TI - NHS direct--via casualty! PMID- 10474381 TI - Care of patients with minor eye trauma. PMID- 10474382 TI - Trauma scoring in A&E. PMID- 10474383 TI - Setting up an ENP service: options, planning and pitfalls. PMID- 10474384 TI - Practices and procedures for universal precautions. PMID- 10474385 TI - A specially dedicated group. PMID- 10474386 TI - Pinpointing the problem of violence. AB - Following a storm of media protest in the UK, health service employers are being forced to re-examine the security measures they implement to protect their staff. PETRA KYDD reviews an RCN survey on violence towards nurses and outlines the benefits of security systems on offer. PMID- 10474387 TI - Principles of wound ballistics. PMID- 10474388 TI - The use of eye drops and ointments in A&E. PMID- 10474389 TI - Fractured zygoma: a case study. PMID- 10474390 TI - Moral, ethical and professional issues in prescribing emergency contraception. AB - Hugh McDonald is an emergency nurse practitioner with the authority to manage a defined group of patients who present to A&E with a specific range of complaints and injuries. Investigation, diagnosis and treatment are carried out independently and include documentation and prescription of specific drugs. PMID- 10474391 TI - Journey from caring to callous. PMID- 10474392 TI - Setting up a paediatric facility. PMID- 10474393 TI - Reducing anxiety in children in A&E. AB - An unfamiliar environment such as A&E can easily provoke feelings of anxiety in an injured child. Julie Morcombe describes a case study of a five year old girl who required treatment for a deep laceration of her arm. PMID- 10474394 TI - Assessing and managing minor burns. PMID- 10474395 TI - Burnout and stress among A&E nurses. PMID- 10474396 TI - Measuring risk and related behaviour with the Behavioural Status Index (BSI): some preliminary psychometric studies. AB - British Special Hospitals are key caregivers for mentally disordered individuals who may display extreme anti-social and challenging behaviour. Assessment of risk and related diurnal behaviour is central to treatment planning for these patients. This paper describes a behaviourally based assessment instrument to assist in the process of the assessment of these 'risky' and related diurnal behaviour. Results of some early data analysis are reported. PMID- 10474397 TI - Influential factors in choosing adult/mental health nursing. AB - This paper describes a comparative survey to investigate the factors influencing twenty undergraduate nursing students on a 'Project 2000 style' course, in their choice of either the adult or mental health branch. The course was structured such that choice of branch was made at the end of the Common Foundation Programme, following experience of each type of nursing. Findings revealed that those choosing the mental health branch were more likely to have made their choice later in the course, and to have changed their mind. Students appeared to have been strongly influenced by positive experiences in mental health placements, both in relation to the work itself, and to the attitudes of ward staff and general atmosphere. These were compared favourably to their experiences in general wards. Reasons for choosing the adult branch included always having wanted to do general nursing and preferring a practically oriented career. Several students were influenced by future career plans for which an RGN qualification was necessary. Both groups had found decision making difficult, especially the mental health branch students who had changed their minds since they began the course. Many students felt that more information and experience, particularly in mental health nursing was needed in the Common Foundation Programme. Suggestions are made in relation to future recruitment to this and other mental health nursing courses. PMID- 10474398 TI - Southeast Asian refugee women and depression: a nursing intervention. AB - Globally, conflicts continue to result in large numbers of refugees and displaced persons, the majority are women. At present, there is scant literature on the mental health status of refugee women following resettlement in countries that grant asylum. We do know that adaptation following migration is a complex cultural, psychological and social process. Some studies have suggested a high prevalence of depression symptoms related to premigration and post-migration experiences. The purpose of this paper will be to describe the mental health status of Southeast Asian (S.E.A.) refugee women in the United States, before home visit interventions by school nurses and bilingual teachers, and at 10, 20 and 33 weeks following the intervention. A comparison group of S.E.A. refugee women, who did not receive the intervention, were evaluated for mental health status on two occasions ten weeks apart. The identified needs and problems identified by the women, the interventions implemented by the school nurses and the success of the interventions will also be discussed. The underlying problem for the majority of women was poverty and social isolation. The study demonstrates that indeed, refugee women in the U.S., are experiencing needs and problems related to basic survival issues in multiple areas of their lives. The findings suggest that home visit interventions by nurses may be a valuable means of reducing depression in S.E.A. refugee women. PMID- 10474400 TI - Public health for children: the Green Paper. PMID- 10474399 TI - Help-seeking pattern of psychiatric outpatients in urban China. AB - The subjects of this study (N = 1297, male = 600, females = 697) were first time adult patients (aged 20 to 59 years) who sought psychiatric treatments at a mental health centre in Shanghai over a period of 12 months. Compared with adult population in Shanghai, the patients were of higher educational levels and were over-represented by the unmarried, separated, or divorced. A higher proportion of the patients were in their early adulthood. The most common presenting problems were sleep disturbance, somatic symptoms, and paranoid ideation. About 25% of the patients took their own initiative to seek psychiatric treatments and more than half (55.2%) were referred by their spouse or relatives. Decision for psychiatric treatments was made after a long delay since the first onset of illness (average = 2.7 years). Consistent with findings reported in other Chinese communities, their paths to seeking psychiatric treatments were diverted primarily through Western-style or traditional Chinese physicians. Stigmatization associated with psychiatric illness and lack of trust in psychiatric services were major deterring factors in seeking psychiatric treatments. PMID- 10474401 TI - The new NHS: the changes, challenges and opportunities. AB - The competition based market system is to be disbanded and will be replaced by more alliances, partnerships and collaborative working practices. Children's nurses are ideally placed to respond to the new NHS call for greater involvement of nurses in the commissioning and service development decision making processes. Primary care groups will build upon the best of existing practice, and offer an opportunities for GPs and nurses working in the community to spread the benefits of their experience more widely. PMID- 10474402 TI - Developing children's nursing through action research. AB - Action research provides an ideal strategy for the integration of theory and practice, as it focuses on the development of theory in practice. Action research is gaining in popularity and acceptance. The collaborative approaches of action research parallel the dominant philosophies of participation and partnership in contemporary children's nursing. Taking the lead in identifying areas of practice for action research is a natural extension of the children's nurse's role as advocate for the child and family. The challenge for the future is to expand the use of action research in children's nursing and to include service users--the children and their families--in the research teams. PMID- 10474403 TI - The 'Bodywise' initiative. AB - Young people want access to good information in suitable settings so they can discuss sexual matters openly. Services are needed to bridge the gap between paediatric and adult health services. Interprofessional collaboration can provide effective, integrated and supportive services for users and providers. Static and outreach services are essential to facilitate young people's access to the services. PMID- 10474405 TI - Assessing the competence of student nurses. AB - Competence is difficult to define. Continuous assessment of practice, which involves reviewing the literature, is complex. The study involved developing a suitable tool for assessing competent practice of student nurses. The problem was investigated using Personal Construct Theory and Repertory Grid Technique. The findings suggest that trained staff assess the socialisation process of becoming a children's nurse rather than the level of competence. PMID- 10474404 TI - Foundations for a healthy future. AB - Health promotion activities with children and young people are important as they take messages about health seriously and can be influential in spreading messages about healthy living to their friends and families. Child health professionals have an important role to play in passing on messages of positive health to children and young people. Peer education is a useful way of passing on messages about health to young people. This article shares examples of three health promotion projects with children in a community trust, looking at asthma, sex education and testicular examination. PMID- 10474406 TI - Dispelling modern day myths about children's pain. AB - Children are still enduring unnecessary pain. Nurses and other health care professionals continue to believe the misconceptions about paediatric pain which contribute to this. These misconceptions have all been shown to have no scientific basis. PMID- 10474407 TI - A right to parenthood. AB - Many people regard parenthood as a 'right' to which they are entitled; this right is open to debate. Infertility can provoke a life-crisis leading to profound feelings of grief, loss and isolation. Each of us, as members of society, must acknowledge the pressure that we place upon individuals to parent, and particularly upon women. Fertility nurses in particular may be caught in the middle of a potentially enormous conflict of interests between the needs and desires of patients and the service provided by the NHS. Child health nurses need to be aware that infertility and the process of fertility treatment may have a significant impact on the parents' relationship with a child born as a result of treatment. PMID- 10474408 TI - Children with AIDS Charity--CWAC. PMID- 10474409 TI - Children--silent consumers of health care. PMID- 10474410 TI - Coping with loss: the development and evaluation of a children's bereavement project. AB - The 'Kingfisher Project' is a community-based programme of support groups for bereaved children. It has been developed by a multi-disciplinary group of workers from health, education, social services and the voluntary sector. Thirty-six children and young people have have been through the first five groups of the programme. Children accessing the service are predominantly those at high risk of developing longer-term problems as a result of bereavement. Evaluation indicates that the children and young people benefit in terms of increased social, behavioural, emotional and physical well-being. Two sub-groups appear to have benefited less: these are children and young people with learning difficulties, and teenage girls. Project evaluation has enabled workers to develop insights into the programme's strengths and weaknesses and has raised questions which merit further exploration. PMID- 10474411 TI - Analysing childhood: a nursing perspective. AB - Childhood is largely socially constructed and is subject to social and cultural change. Childhood is often characterised by immaturity and dependency, which may hamper the full recognition of children's rights. Children's nurses, as children's advocates, should develop a critical awareness of the concept of childhood. Children's nurses should also consider undertaking phenomenological research with children, as a means of identifying the needs of children. PMID- 10474412 TI - The partnership myth? Examining current opinions. AB - There is a need for further clarification of the concept of partnership. A philosophy of 'family-centred care' will require negotiation with, and commitment from, all concerned with care delivery in the ward and beyond. Paediatric nursing needs to adopt a flexible and non-judgmental approach in order to facilitate the needs of all families. The concept of 'partnership in care' is of benefit to families caring for sick children in the context of today's economic and political climate. PMID- 10474413 TI - Equal opportunities, welfare and child health: a review. AB - Many families from ethnic minority backgrounds are disadvantaged. These families have poorer access to health care provision. Social factors such as poor housing are detrimental to both child health and child safety. Nurses from minority groups remain under represented within the NHS. PMID- 10474414 TI - Influences on nurses' perceptions of children's pain. AB - Individuals have their own perceptions of pain. Nurses' do not always perceive their patients' pain. There are a number of factors which influence nurses' perceptions of their patients' pain. Further research is needed in this area. PMID- 10474415 TI - The theory-practice gap and student nurses. AB - The theory-practice gap in nursing is a multi-factorial problem. Different types of knowledge are used in nursing; some have more credence and value than others. Learners are often underestimated by clinical staff, they can provide sophisticated care while being relatively inexperienced. 'Reality shock', is perhaps due to a lack of student's emotional readiness to deal with the daily stresses that nursing entails. PMID- 10474416 TI - The Child Growth Foundation. PMID- 10474417 TI - Nurses' personal pain experiences and their pain management knowledge. AB - BACKGROUND: This study examined nurses' personal experiences with pain and whether nurses' personal pain experiences were associated with initial pain management knowledge and ability to learn more about pain management. METHOD: This descriptive correlational study was a secondary analysis of a study that used a one-group pretest-posttest design to examine the effect of a pain management education program on nurses' pain management knowledge. The sample consisted of 177 nurses who answered yes to having had a personal pain experience requiring treatment. RESULTS: Nurses reported 13 types of pain, with pharmacological interventions comprising 94.4% of the pain treatments. Nurses' previous personal pain experiences were negatively related to their initial pain management knowledge (r = -.23, p < .01). CONCLUSION: Nurses' personal pain experience may impact their learning of pain management knowledge and should be considered when designing pain management education. PMID- 10474418 TI - Hardiness and anxiety as predictors of academic success in first-year, full-time and part-time RN students. AB - BACKGROUND: The levels of hardiness and anxiety and their relationship with academic success were examined using the Personal Views Survey and the State Trait Anxiety Inventory. METHOD: The instruments were administered to a nonprobability convenience sample of 41 full-time and part-time RN students enrolled in a BSN completion program. RESULTS: Participants perceived themselves as possessing moderately high levels of hardiness (mean = 72.90) and low levels of anxiety (state mean = 39.33; trait mean = 40.18), but for some, these characteristics did not relate to academic achievement. Statistical significance was set at .05. CONCLUSION: These findings indicate how hardiness and anxiety can affect academic success, particularly in adult students, and provide some direction for educators and administrators in maximizing educational endeavors. PMID- 10474419 TI - Continuing professional education and interacting variables affecting behavioral change in practice: instrument development and administration. AB - BACKGROUND: Instruments were developed to measure variables identified by Cervero that evaluate the complex processes and effectiveness of continuing professional education. METHOD: Content validity was established with a panel of experts. Four studies were conducted to determine reliability. Data were merged to conduct factor analysis. RESULTS: Reliability was confirmed across studies using Cronbach's alpha. Stability was confirmed in a test-retest study. Factor analysis confirmed instrument items measure the concepts of Cervero's Model. CONCLUSION: The reliable and valid instruments may be useful for continuing education and staff development nurses to measure the effectiveness of education on behavioral change in practice. PMID- 10474420 TI - Assessment of nurses' continuing education needs in Jordan. AB - This study assessed the learning needs of hospital nurses in northern Jordan to develop programming recommendations for future continuing education (CE) efforts. Although few CE programs exist, a response rate of 85% (n = 171) suggested these nurses have important learning needs. Nurses identified eight priority content areas: critical care, management of cardiac arrest, emergency room nursing, delegation and accountability, power and authority, teaching innovations, seminar management, and patient education. The findings showed that almost two thirds of subjects (n = 110) had not attended a CE program in the previous year, and very few (n = 8) had read nursing journals to supplement their nursing knowledge. Although data suggested lack of motivation to participate in CE programs was a frequent problem, they also indicated CE participation would increase if participants could earn some type of credit for this additional education. PMID- 10474421 TI - Self-paced: the right pace for staff development. AB - BACKGROUND: Limited budgets and increasing training demands on staff development educators require creativity and innovation in providing cost-effective training alternatives. Self-paced learning modules meet these criteria and minimize use of staff time. METHOD: This article presents a step-by-step guide for the design and implementation of self-paced learning modules. RESULTS/CONCLUSION: By following the suggested guidelines, even novice staff development educators can assess, design, and implement cost-effective training alternatives for their organizations. PMID- 10474422 TI - Clinical supervision. The feeling of safety engendered by supportive supervision. PMID- 10474423 TI - Primary care groups. Commissioning effectively. PMID- 10474424 TI - Critical appraisal. 2. Searchable questions. PMID- 10474425 TI - Education for practice. Whose lifelong learning is it, anyway? PMID- 10474426 TI - Clinical governance. 1. Organisational effectiveness--1. PMID- 10474427 TI - Healthcare open learning. Factors that affect how and what we learn. 2. Curriculum models. PMID- 10474428 TI - Education for practice. The search for a credible model for the teacher/lecturer in practice. PMID- 10474429 TI - Healthcare open learning. Empowerment through practice-based learning. 1. Power and socialisation. PMID- 10474430 TI - Clinical supervision. Supportive supervision as a means of enabling self awareness. PMID- 10474431 TI - Critical appraisal. 3. Searching for the evidence. PMID- 10474432 TI - Primary care groups. Lay involvement in primary care. PMID- 10474433 TI - Higher level practice. PMID- 10474434 TI - Education for practice. Problem-based learning: giving control to the student. PMID- 10474435 TI - Learning in clinical practice. 1. Preparing to be a mentor. PMID- 10474436 TI - Electronic records and informal networks. PMID- 10474437 TI - Clinical supervision. Building professional relationships. PMID- 10474438 TI - On the job. The clinical nurse specialist: leading the way to quality care. PMID- 10474439 TI - [The mandates of ethics in nursing]. PMID- 10474440 TI - [Ethical aspects of enteral feeding]. PMID- 10474441 TI - [May the dignity of the disabled be infringed upon?]. PMID- 10474442 TI - [Teacher training in university courses]. PMID- 10474443 TI - [The situation of female migrants with HIV/AIDS]. PMID- 10474444 TI - [The German Nursing Society in 1997 and 1998--a review]. PMID- 10474445 TI - [Diagnosis related groups and nursing: what effect on the quality of care? Ethical-deontological implications]. AB - Each health system adopts distribution patterns of its available resources, establishing the kind, the amount and the quality of health care accessible for individuals. In this way people's state of health is directly affected. Nurses must know moral implications of these choices and have to be able to interpret them not only at a political/institutional level, but also at an operative one, inside each care intervention. Such a behaviour is determined by the possibility of conflict between the administration choices and the guarantees nurses are engaged to offer to persons in need of nursing care. Its moral aim is the achievement of the best nursing care to individuals. PMID- 10474446 TI - [Diagnosis-related groups and nursing. Effect on the quality of nursing care]. AB - In this article we consider different DRG's variables that influence the performance of nurses. We analyze particularly the subsystems defined by Kast and Rosenzweig. PMID- 10474447 TI - [The power of nursing versus minimal nursing data]. AB - This article reports the Belgian experience of the Nursing Minimum Data Set (NMDS), in order to make the nurse work visible. The collected data are forwarded to the Health Ministry and, together with the DRGs, constitute the base for the determination of hospitals financing budget in Belgium. PMID- 10474448 TI - [The mask and the face of a profession]. AB - These pages criticize those forms of extreme medicalization that support a fragmented and sectional health anthropology. They declare the importance of the nursing profession as a methodologic discipline based on the help relationship. Furthermore, these pages call back towards social and anthropologic "vocation", like in the beginning. Such vocation allows to approach the ill patient in his/her totality. It also antagonizes contemporary medical philosophy, which has produced the patient-machine standing on a functionalist and purely instrumental business culture of therapy and care. PMID- 10474449 TI - [The patient as a person in the hospital as a business]. AB - The ethical and bioethical questions are assuming an increasing importance within the changes that characterize the health field. Nursing, as other professions, examines the relationship between ethics and professional practice. Ethics is often thought of as an element belonging to the real nature of the profession, but it is necessary to know which exigence gives rise to this presumption. The core of all the different ethical conceptions existing to day is always the concept of "the person", that determines different behaviours. To be able to develop ethical responsibilities it is essential to know who the person is. PMID- 10474450 TI - [Studies carried out in the province of Nuoro on treatment modalities for oncologic pain during the terminal phase]. AB - The purpose of this first level research is to analyze the organization of the treatment of oncological pain in one Italian province. For the data collection a questionnaire was submitted in the Intensive Care Unit to the responsibles and to the staff in the public hospital structures of the six USL of the Nuoro's province. Results prove that the motivation of the staff and their problem sensitivity, are inadequate to guarantee an effective answer to the patient's need. The main aspects are: legal aspect, concerning the district level interventions ache care formation aspect for registered nurses the necessity to adopt quality evaluation methods. PMID- 10474451 TI - [The psychodynamic nursing theory by Hildegard E. Peplau]. AB - Peplau's theory is one of the early Nursing theories, published in 1952. The nurse-patient relationship consists of four steps (orientation, identification, development and conclusion). In these steps nurse could have the role of foreign, reliable person, teacher, guide in nursing care, substitute and consultant. Nurse patient relationship is influenced by psychobiological experiences (needs, frustrations, conflicts and anxiety) which need dynamism. Peplau thinks that Nursing care is an important opportunity for nurse because she can help patient to complete the infancy psychological tasks (learning to rely on other people, learning to show satisfaction, self-identifying, and developing ability in sharing) if these are not completed. For these reasons Nursing, by Peplau, is a maturation strength of civilization. PMID- 10474452 TI - [Using Peplau's theory in the nurse-patient relationship]. AB - In caring for a person suffering from depression, Peplau's theory of "Interpersonal Relations" was found to be both effective and representative of what psychiatric nurses do. Peplau's work, initially published in 1952, continues to offer a major contribution to nursing knowledge and deserves the significance ascribed to it in the 1990s. Below, a report on the learning experience for both the patient and the carrier. PMID- 10474453 TI - [Contribution to the International Congress of Nurses: acts of violence and accidents]. PMID- 10474454 TI - [The screech owl. Needlestick injury]. PMID- 10474455 TI - [The council of Europe seeks a promise from member states about their position on nursing research]. PMID- 10474456 TI - [AIDS and the new therapies: course of research]. AB - The Authors notice the impact of AIDS on all fields of the social life. After outlining a short story of the disease and of the different therapies applied till now, they review the new ones and point out the necessity to provide nurses with specific and continuous education. PMID- 10474457 TI - [Care of persons with AIDS: choosing home care]. AB - The main care perspectives for the persons with AIDS are analyzed in the article. A particular attention is given to some conceptual aspects of home care as a priority choice resulting from the recent developments of both the nursing research and the Italian regulations. PMID- 10474458 TI - [Quantitative and qualitative nursing research]. AB - The aim of this article is to open a discussion on Nursing research methods. Authors give some thoughts on qualitative nursing research and underlining the difference between positivistic and teleological vision. Relationship between inductive and deductive thinking is discussed. PMID- 10474459 TI - [European Health Committee. Nursing research. Report and recommendations]. PMID- 10474460 TI - [Nurses' experience of violence in psychotic crisis: an example of qualitative research]. AB - This study indagates into the personal experience of a nurse caring for an aggressive patient in acute psychotic crisis and gives an example of qualitative research. PMID- 10474461 TI - [Analgesic therapy service: analysis of one year's activities]. AB - In this article we describe the activities an anthalgic therapy service has carried out in one year. Considering the peculiarity of the involved pathologies, we point out the necessity of both a continuous education and a service structured in such a way as to meet the patient needs. PMID- 10474462 TI - [The role of the professional nurse in the application of natural medicine techniques]. AB - The increasing attention addressed to the natural medicine is pointed out in this article. The Author emphasizes that in this field nurses play an important role of mediators helping the patient for the transition from the traditional medicine and defines the kind of natural techniques nurse may apply. PMID- 10474463 TI - [Accreditation visits: role of the evaluator]. AB - Accreditation visits are used by Italian Health "Aziende" as ways how to evaluate themselves in comparison with the pre-definite requirements. They can be used to be allowed to offer health services when there are the necessary qualifications. The "judge" or "expert" should support the "Aziende" in order to help in getting knowledge and qualification to "credit" the organization. We want to examine this subject "new" to the Italian Health System through the experience of the Ferrara Hospital making the Nurse aware of the importance of taking part in it. Therefore the Nurse will have to accept the new challenge and new development objectives. PMID- 10474464 TI - [The screech owl. Wondering about the nature of nursing relationships]. PMID- 10474465 TI - Comparative objective and subjective analysis of temporalis tendon and microneurovascular transfer for facial reanimation. AB - The aim of this study was to compare objectively and subjectively the results after temporalis tendon and microneurovascular transfer for reanimation of the paralysed face. For the objective analysis, measurements of the distances between static and dynamic points in three-dimensional space were performed at rest and during smiling. The patients were assessed subjectively by non-professionals scoring videotapes. The range of motion of the oral commissure on smiling was 1.7 mm (0.8, mean and SE) after tendon and 5.5 mm (1.6) after microneurovascular transfer (P < 0.05) without showing teeth, and 0.6 mm (1.3) vs 3.6 mm (2.4, ns) with showing teeth. Compared with the healthy side, the excursions reached 6% (16) after tendon and 71% (26, P < 0.05) after microneurovascular transfer for smiling without, and -4% (14) vs 19% (14, ns) for smiling with showing teeth, respectively. Virtually no difference was found in the measurements at rest as well as in the subjective evaluation. Swelling of the cheek (28%) and tethering of the skin (24%) were shown to be the most disturbing symptoms after microneurovascular transfer. Better excursion and symmetry of the oral commissure were achieved by microneurovascular reconstruction for voluntary smile. However, the aesthetic outcome after microneurovascular transfer is markedly impaired by chronic complications including swelling of the cheek and tethering of the skin, which were not detected by the three-dimensional measurement technique used. We conclude that the aesthetic appearance should be considered equally important as the range of motion in the reanimated face. PMID- 10474466 TI - The removal of unwanted hair using a ruby laser. AB - A ruby laser has been developed to remove unwanted hair. Melanin within the hair is used as a natural chromophore. It is postulated that photothermal damage destroys the hair itself and also key cells surrounding the hair follicle to prevent regrowth. A prospective study of laser depilation in 116 patients or 175 sites was performed over a period of 18 months. All the patients had tried other methods of hair removal and found them to be unsatisfactory. Hair counts (follicles/cm2) were used to judge the outcome. The mean follow-up time was 23.25 weeks (range 12-76 weeks). After a mean number of treatments of 1.92 (range 1-20) there was a 56.4% reduction in hair density. Comparing pre- and post-treatment hair density, there was a highly significant reduction (paired t-test: P < 0.00001). Laser removal of hair is now a realistic treatment option. PMID- 10474467 TI - Intralesional photocoagulation of vascular anomalies of the tongue. AB - Vascular anomalies remain a challenge for patients and reconstructive surgeons. Recently, promising results have been reported using intralesional photocoagulation (ILP) to treat large, deep vascular lesions. We report our experience in treating 12 patients with vascular anomalies of the tongue (10 venous malformations and two haemangiomas) using intralesional photocoagulation. All patients were treated with a Nd:YAG (neodymium:yttrium-aluminium-garnet) (1064 nm) laser (Sharplan, Inc., NJ, USA) delivered with a 600 microns optical fibre. Laser power was set at 7 or 10 W, delivered with a pulse duration of 10 s. Laser energy was delivered to all areas of the vascular lesion. Heat, visible shrinkage and firmness of the lesion signalled the end-point of treatment. The mean follow-up period was 9.5 months (range 3-20 months). All patients demonstrated improvement as judged by a clinical assessment of the reduction in lesion size (mean reduction = 87%, range 60-100%). Three patients (25%) had complications attributed to photocoagulation. Energy delivered too superficially resulted in ulceration or scarring. These complications should be avoidable if this potential for harm is kept in mind. PMID- 10474468 TI - Endoscopic extraction and curettage of epidermal cysts on the face. AB - Endoscope-assisted extraction and curettage of epidermal cysts on the face has been successfully performed in five patients. An incision was made in the external acoustic meatus close to the tragus. Another incision was made behind the earlobe. Through one incision, the 30 degrees, 4-mm-diameter endoscope was inserted. The other incision was used for the insertion of forceps, scissors and curettes. All of the cysts were extracted without additional incisions. All of the patients were satisfied with the outcome. This method is applicable in patients to reduce visible scarring on the cheek. PMID- 10474469 TI - The sensate free superior gluteal artery perforator (S-GAP) flap: a valuable alternative in autologous breast reconstruction. AB - The superior and inferior myocutaneous gluteal free flaps have been considered as valuable alternatives to the latissimus dorsi or TRAM flap since 1975. The superior gluteal artery perforator (S-GAP) flap is the ultimate refinement of this myocutaneous flap as no gluteus maximus muscle is harvested. The flap is vascularised by one single perforator originating from the superior gluteal artery. This study summarises the prospectively gathered data on 20 free S-GAP flaps used for breast reconstruction in 16 patients. Immediate reconstruction was performed in six breasts and delayed in 14 breasts. Mean follow-up was 11.1 months. Two risk factors, Raynaud's disease and radiotherapy, were the cause of flap revision in two different patients. Total flap loss occurred in one case. Partial flap loss was not observed and a small area of fat necrosis was diagnosed by mammography in one other patient. All flaps were anastomosed to the internal mammary vessels at the 3rd costochondral junction. The anatomy of the sensate nerves of the S-GAP flap is described. Two nervous repairs provided early sensory recovery. The free S-GAP flap has become my personal second choice for autologous breast reconstruction after the DIEP (deep inferior epigastric perforator) flap. The S-GAP flap is indicated in patients with an asthenic body habitus or with excessive abdominal scarring. The advantages are the abundance of adipose tissue in this area even in thin patients, a long vascular pedicle, a hidden scar, improved projection of the reconstructed breast compared to the DIEP and TRAM flaps and the preservation of the entire gluteus maximus muscle. The donor morbidity is extremely low. PMID- 10474470 TI - Topical application of DMSO as an adjunct to tissue expansion for breast reconstruction. AB - The aim of this study was to evaluate the effectiveness of topical 60% dimethyl sulphoxide (DMSO) prior to tissue expansion for breast reconstruction in reducing expander pressure and length of treatment. Forty patients undergoing immediate breast reconstruction with tissue expanders following modified radical mastectomy were consecutively divided into two groups of 20 subjects each: group A (control) and group B (experimental). Patients from group A underwent traditional tissue expansion, while, to patients of group B, before each filling session, topical 60% DMSO was applied to the skin overlying the expander by soaking-wet surgical sponges left in place for 30 min. The average inflated volume was 395 cc (range 250-580 cc) in group A, and 410 cc (range 240-620 cc) in group B; no statistically significant difference was found between these values. A statistically significant difference was found between the expansion time of group A (mean 6 sessions; range 4-10 sessions with one filling session per week) and group B (mean 4 sessions; range 2-6 sessions with one filling session per week). A statistically significant difference was also found between the average inflated volume per session of group A (90 cc; range 60-130 cc) and group B (120 cc; range 90-160 cc). According to the data obtained by tonometry, a statistically significant difference was found between the average pre-filling and post-filling pressures of group A (28.4 and 66.5 mmHg, respectively) and group B (22.1 and 64.3 mmHg, respectively). Clinical benefits of pre-filling topical applications of 60% DMSO observed in our trial were an enhancement of the total volume inflatable per session and a significant shortening of the total expansion period. PMID- 10474471 TI - A qualitative study of patients' views on the effects of breast-reduction surgery: a 2-year follow-up survey. AB - The objectives of this study were to discover the views of patients about the effects of breast-reduction surgery carried out 2 years previously, to detect any change from perceptions at 3-6 months after surgery, and to determine whether the benefits of this operation are maintained long term. Qualitative research methods were employed, comprising: (a) an open-format survey of opinions; (b) semi structured telephone interviews with a smaller number of patients; and (c) assessment of self-concept using a well-known scalar measure (the Rosenberg Self esteem Scale). The subjects were 93 patients treated at the regional Plastic Surgery Service in Salisbury, who had previously participated in a quantitative study at 3-6 months after surgery. Sixty patients responded to the 2-year follow up. Benefits of breast reduction most valued by patients did not change significantly with time and were: relief of pain and discomfort, which led to increased physical activity and better general health; greatly increased choice and fit of attractive clothes and underwear; improved personal and social life, leading to enhanced relationships with partner or friends; and greatly improved self-confidence in all areas of life. The interaction of all these factors led to improved self-image and improved quality of life. The main disadvantage of the operation for a small number of patients was the persistence of painful, disfiguring scarring which in two cases had a detrimental effect on social and personal relationships and led to a deterioration in quality of life. Improvement in self-esteem after surgery was maintained in 55 out of the 60 2-year responders. The results indicate that breast reduction confers significant health gains which are maintained in the long term. PMID- 10474472 TI - Progressive skin extension: clinical and histological evaluation of a modified procedure using Kirschner wires. AB - We describe a modification of a skin traction system for progressive skin extension before excision of skin lesions. The traction device consists of Kirschner wires and plastic straps. It results in more homogeneous traction and greater sparing of healthy skin as compared with the earlier silicone bar method. Moreover, the procedure results in lower rates of infection, skin necrosis and breakage of the cutaneous bridges. Histological changes to the treated skin area are similar to those obtained with skin expansion procedures. PMID- 10474473 TI - The pathogenesis of local recurrence of melanoma at the primary excision site. AB - Local recurrence of melanoma at the primary excision site may imply that the primary excision was incomplete or 'inadequate', and that the recurrence was due to retained primary melanoma cells or occult microsatellites in the adjacent tissue. Pathologists frequently report these tumours in the scar as recurrent or residual melanoma, without further qualification, apparently without considering the possibility that they may be metastases and manifestations of systemic disease. In this study, 17 of 19 cases of locally recurrent melanoma at the primary excision site showed the histological features of metastasis rather than residual incompletely excised primary melanoma. Because the prevention of local recurrence is the main reason given in recommendations for wide excision of melanoma beyond complete excision of the primary tumour itself, it is essential that surgeons and pathologists should classify these neoplasms precisely as either persistent incompletely excised primary melanoma or metastatic melanoma. PMID- 10474474 TI - Does aspirin affect the outcome of minor cutaneous surgery? AB - The use of regular aspirin as an antithrombotic agent has become prevalent in the elderly population. The belief that operative bleeding complications are more common in those patients taking regular aspirin has commonly led to its withdrawal 5-10 days prior to minor dermatologic plastic surgery. Few studies exist to examine the true effects of aspirin on outcome in these patients. All patients presenting for minor dermatologic plastic surgery in a 6-month period were prospectively studied. Two age-matched cohorts were identified: 52 patients taking regular aspirin, and 119 patients taking no aspirin. The incidence of minor and significant bleeding complications was recorded. There was no significant difference in the incidence of minor, significant or total complications between the two groups (Fisher's Exact test, P = 1.00). It is concluded that it is unnecessary to stop aspirin before minor dermatologic plastic surgery. PMID- 10474475 TI - Phalloplasty in female-to-male transsexuals using free radial osteocutaneous flap: a series of 22 cases. AB - From July 1994 to December 1996, 22 cases of primary female transsexualism received phalloplasty using free radial forearm osteocutaneous flaps in The Veterans General Hospital, Taipei, Taiwan. Follow-up period ranged from 6 months to 35 months. No complete flap loss and one partial loss (10% reduction) were noted after operation. The urethrocutaneous fistula rate was 40.9%, needing fistula repair surgery. Urethral stricture was noted in three cases (13.7%). Significant donor forearm morbidity was noted in two cases (9.1%) including one who suffered from radius bone fracture. All preserved 'clitorises' had intact erotic sensation and all neophalli gained protective sensation within 9 months. No 'penile fracture' was noted. In nine cases that have regular sexual activities, the sexual performance was rated as satisfactory. We share our experience and recommend that the free radial forearm osteocutaneous tubing flap is a promising choice for phalloplasty in female-to-male transsexuals. PMID- 10474476 TI - Transfer of a pedicled venous flap containing perivenous areolar tissue and nerve: an experimental study. AB - In this paper, we evaluated experimentally the feasibility of transferring a pedicled venous flap with perivenous areolar tissue into an ectopic site and investigated the role of each vein, the perivenous areolar tissue and nerve in the viability of pedicled venous flaps. Three groups of flaps were created using a rabbit-ear model: Group 1 (n = 11), containing perivenous areolar tissue and a draining vein; Group 2 (n = 10), perivenous areolar tissue, a draining vein and nerve; Group 3 (n = 10), only perivenous areolar tissue. Macroscopic, angiographic and histological assessments were performed 14 days after transfer. The total survival areas including superficial necrosis of Groups 1, 2 and 3 were 52.7%, 82.5% and 0%, respectively. There were significant differences in mean survival area and mean total survival area including superficial necrosis between Group 1 and Group 2 (P < 0.05). In contrast, all flaps in Group 3 became necrotic. These data suggest that a preserved vein is the necessary condition, and additional arterial flow through the preserved nerve and a small vascular network within the areolar tissue may play an important role in flap survival. PMID- 10474477 TI - Botulinum toxoid in the management of gustatory sweating (Frey's syndrome) after superficial parotidectomy. AB - Botulinum toxin has been successfully used to treat Frey's syndrome occurring in a 31-year-old patient following superficial parotidectomy for pleomorphic adenoma. An initial injection of 7.5 U (0.3 ml over 6 cm2 of cheek) resulted in 3 months' resolution of gustatory sweating and flushing and a second injection 12 months' symptomatic improvement. The symptoms recurred after further facial surgery. PMID- 10474478 TI - A functional variant of lower lip reconstruction. AB - A functional variant of lower lip reconstruction is reported, in which a nasolabial skin flap was combined with upward transposition of the inferior part of the orbicularis oris muscle in a 'bucket-handle' fashion. The tongue was split to provide both an intraoral lining to the lip and a new vermilion. PMID- 10474479 TI - Pulsed dye laser treatment of telangiectasia after radiotherapy for breast carcinoma. AB - Telangiectasia formed following exposure to X-ray irradiation have been reported to respond well to the pulsed dye laser system. We present the case of a 34-year old woman with extensive post-radiotherapy skin telangiectasia of the chest wall and axilla who was treated with six sessions of pulsed dye laser treatment, with a considerable improvement in appearance. We recommend the pulsed dye laser as an option in the treatment of post-radiotherapy telangiectasia. PMID- 10474480 TI - Lipid emulsification after implant rupture. PMID- 10474481 TI - Perioperative improvement in lung function during reduction mammoplasty. PMID- 10474482 TI - Weight discrepancies in surgical and pathological specimens from breast reductions. PMID- 10474483 TI - Cutting diathermy for removal of implants--reply. PMID- 10474484 TI - Preoperative counselling for breast augmentation. PMID- 10474485 TI - The use of blood as a marker on the lips: an aid to reconstruction. PMID- 10474486 TI - Simulated interactive local flaps: operating room models for surgeon and patient alike. PMID- 10474487 TI - The microvascular simulator. PMID- 10474488 TI - Aspirin and surgical bleeding. PMID- 10474489 TI - Long-term survival of mandibular osteosarcoma. PMID- 10474490 TI - Variation between experienced observers in the interpretation of accident and emergency radiographs. AB - Skill mix and role extension initiatives have highlighted the difficulty of establishing quality standards for the accuracy of plain film reporting. An acceptable performance might be one which is indistinguishable from that of a group of experienced consultant radiologists. In order to assess the feasibility of setting such a standard, the variation between experienced observers must first be established. This study examines the variation found between three observers with the three major types of plain film examination. 402 plain film examinations (205 skeletal, 100 chest and 97 abdominal) performed on accident and emergency patients were reported retrospectively and independently by three experienced radiologists. The clinical data supplied on the request cards were available to the readers. Each examination was categorized by each reader as being normal, as showing significant abnormality relevant to the current clinical problem, or as showing insignificant or irrelevant abnormality. Concordance between all three readers was found in 51%, 61% and 74% of abdominal, chest and skeletal radiographs, respectively. Weighted kappa values confirmed that the level of agreement between pairs of observers was higher with skeletal radiographs (kappa w = 0.76-0.77) than with chest (kappa w = 0.63-0.68), or abdominal (kappa w = 0.50-0.78) examinations. However, the frequency of major disagreements (at least one reader reporting "normal" and one reporting "relevant abnormality") was similar for abdominal (11%), chest (12%) and skeletal (10%) radiographs. When the reports were reclassified into only two groups--either significantly abnormal or not--pairs of observers disagreed on 9-10% of skeletal, 11-19% of chest and 8-18% of abdominal cases. The average incidence of errors per observer was estimated to be in the range 3-6%. The magnitude of interobserver variation in plain film reporting is considerable, and must be taken into account when designing assessment techniques and setting quality standards for this activity. PMID- 10474491 TI - Reliability of radiological measurements in the assessment of hip dysplasia in adults. AB - Radiographic measurements are commonly used to quantify the treatment results of hip dysplasia and assess further need of operative treatment. We investigated the interobserver and intraobserver reliability of the commonest radiographic techniques in the assessment of hip dysplasia in skeletally mature adults. Three observers independently analysed 100 hip radiographs of patients with hip dysplasia aged between 16 and 32 years. We measured centre-edge angle of Wiberg, acetabular angle of Sharp, acetabular index of the weightbearing zone, acetabular index of depth to width, ACM-angle, MZ-distance, acetabular head index, lateral subluxation and neck-shaft angle. In addition, the radiographs were reviewed a second time 3 months apart by two of the observers to assess intraobserver reliability. We found a high correlation (intraclass correlation coefficient) for interobserver reliability (0.76-0.87) and intraobserver reliability (0.70-0.92) for all radiographic measurements except acetabular index of depth to width, ACM angle and MZ-distance. Depending on the clinical question we therefore recommend the use of one of the reliable measurements to assess the radiograph of a dysplastic hip. PMID- 10474492 TI - A radiographic method for assessing lung area in neonates. AB - The aim of this study was to determine whether computer assisted analysis of lung area on the chest radiograph reliably predicted lung volume in neonates. Anteroposterior chest radiographs taken for clinical purposes were scanned and analysed using a Power Macintosh computer with a Wacom A5 Ultra Pad and NIH image software. The cardiac, mediastinal and thymic densities and areas of perihilar and lobar consolidation were subtracted from the thoracic area to give the lung area. This was compared with lung volume, assessed by measurement of functional residual capacity (FRC), within 1 h of the chest radiograph being performed. 50 infants, median gestational age 30 weeks (range 24-43) were studied. Their median lung area was 11.23 cm2 (range 0.82-28.53) and lung volume 28 ml (range 3-103). The intraobserver and interobserver coefficients of repeatability of lung area were 1.0 cm2 and 1.06 cm2, respectively. Lung area correlated significantly with FRC (r = 0.60, p < 0.0001). It is concluded that computer assisted analysis of the chest radiograph lung area is a reliable method of assessing lung volume in neonates. PMID- 10474494 TI - How sensitive is ultrasound in the detection of renal scars? AB - The English language medical literature was reviewed to determine the strength of the published evidence for the assertion that dimercaptosuccinic acid scintigraphy (DMSA) is superior to ultrasound (US) in the detection of established renal scarring in children. The MEDLINE database was used to identify papers published between 1985 and 1997 that claimed to be concerned with the detection of scars, and contained sufficient information to permit calculation of the sensitivity of US relative to DMSA. Only 10 studies were identified. The sensitivity of US for scarring, using DMSA as a gold standard, ranged from 37% to 100%, and its specificity from 65% to 99%. These wide ranges mean that evaluation of the role of US in the detection of scarring remains controversial. All papers contained methodological flaws. Allowing for these, the sensitivity of US appears to be acceptable. Further research that avoids these methodological problems is required. PMID- 10474493 TI - Pulmonary tuberculosis: radiological features in west Africans coinfected with HIV. AB - A retrospective study was performed to document and compare the radiological appearances of newly diagnosed pulmonary tuberculosis (PTB) in groups of West African patients with (n = 86) and without (n = 106) human immunodeficiency virus (HIV) coinfection. Analysis of chest radiographs showed that the HIV-positive group had less consolidation (mean 3.1 zones vs 3.7 zones; p < 0.05), less apical involvement (64.0% vs 85.5%; p < 0.001), less bronchopulmonary spread (27.9% vs 58.5%; p < 0.001), less volume loss (53.5% vs 76.4%; p < 0.001) and less pleural thickening (46.5% vs 61.3%; p < 0.05) compared with the HIV-negative group. However, HIV-positive patients more commonly had pleural effusions (17.4% vs 6.6%; p < 0.05) and lymphadenopathy (9.3% vs 1.9%; p < 0.05). Previous studies on this subject from sub-Saharan Africa have focused either on selected patient groups likely to have more advanced immunosuppression or on smear-positive cases only, or where there has been only limited radiological documentation. This study suggests that the highly significant differences that exist may not be as frequent as previously shown. The lower frequencies of bronchopulmonary pattern of consolidation and pleural thickening in HIV-positive subjects have not previously been documented. The possible reasons for the altered radiographic appearance of PTB in HIV positive subjects are discussed. PMID- 10474495 TI - MR features of the denervated tongue in radiation induced neuropathy. AB - The MR features of the denervated tongue have been described following nerve injury from radical neck dissection and tumour invasion. The purpose of the study was to determine whether similar features are present in the tongue following radiation induced neuropathy (RIN). The clinical records and MR images of 12 patients with RIN of the hypoglossal nerve were reviewed retrospectively. T1 weighted SE images were performed in 12, T2 weighted TSE images in 11, fat suppressed images in 10 and contrast enhanced T1 weighted images in nine patients. The denervated tongue revealed "oedemalike" changes in five, fatty infiltration in six, atrophy in 11 and pseudohypertrophy in one patient. Abnormal enhancement was not identified, and in five patients the signal intensity was normal on all sequences. The oedemalike changes, fatty infiltration and normal signal intensity were seen 2-48, 2-48, and 6-63 months, respectively, after the onset of RIN. In conclusion, there was no discernible relationship between the duration of RIN and the MR appearance of the denervated tongue. Oedemalike changes, previously described in the acute/subacute phase of denervation, were also seen in long-standing disease and there was no associated abnormal enhancement in any case. Furthermore, the signal intensity may be normal, the MR diagnosis relying on asymmetry of the size of the tongue. It is postulated that radiation causes incomplete and ongoing damage of the nerve, the course of which is unpredictable. PMID- 10474496 TI - Cross-calibration, precision and patient dose measurements in preparation for clinical trials using dual energy X-ray absorptiometry of the lumbar spine. AB - Large, multicentre clinical trials using DXA to monitor bone density following intervention are now common. At the same time, several different bone densitometers and calibration phantoms are currently in use. The aim of this study was to document the technical information required on cross-calibration of equipment, reproducibility and patient dose before commencing a multicentre clinical trial. To this end, we obtained an in vitro and in vivo cross calibration of two machines (a Hologic QDR 2000 and a Lunar DPX-L) that were not significantly different. Interobserver and intraobserver precision, and radiation dose were also measured and three commonly used phantoms assessed for their usefulness in cross-calibration and quality assurance. Measured in vitro precision on the two machines (0.3-0.7%) was better than that specified by the manufacturers. In vivo precision was worse (1.4-2.1%), as might be expected in patients with reduced bone mass. Mean entrance skin radiation doses on each machine were 280 microSv for the QDR 2000 and 38 microSv for the DPX-L. No one phantom is ideal, but the European Spine Phantom or Lunar Aluminium Spine Phantom will provide an adequate cross-calibration for a clinical trial. This study demonstrates that an adequate cross-calibration can be obtained for use in groups of patients and that the equipment used is reproducible with a low radiation output. PMID- 10474497 TI - Fetal position and size data for dose estimation. AB - In order to establish both positional and size data for estimation of fetal absorbed dose from radiological examinations, the depth from the mother's anterior surface to the mid-line of the fetal head and abdomen were measured from ultrasound scans in 215 pregnant women. Depths were measured along a ray path projected in the anteroposterior (AP) direction from the mother's abdomen. The fetal size was estimated from measurements of the fetal abdominal and head circumference, femur length and the biparietal diameter. The effects of fetal presentation, maternal bladder volume, placenta location, gestational age and maternal AP thickness on fetal depth and size were analysed. The fetal position from the anterior surface of the mother's abdomen is shorter for posterior placenta and empty bladder volume, but longer for anterior placenta and full bladder volume. Mean fetal depth (MFD) observed for all bladder volumes, fetal presentations and placenta locations increased from 6.5 +/- 0.5 cm to 10.2 +/- 0.7 cm over the duration of pregnancy. Similarly, mean fetal skull depth (FSD) increased from 6.6 +/- 0.6 cm to 9.8 +/- 0.6 cm over the period of pregnancy, but only from about 6.6 cm to 7.8 cm over the period (8-25 weeks) when damage to the developing brain has been observed to result in mental retardation. Using the range of mean fetal depth (4.7-13.9 cm) observed in this study and depth dose data at 75 kVp and 3.0 mmAl half value thickness (HVT), fetal absorbed dose would be overestimated by up to 66% or underestimated by up to 77% if the mean value of MFD (8.1 cm) is used rather than actual individual values. These errors increase with lower tube potential and filtration up to over 90% overestimation and up to 100% underestimation at 60 kVp and 1.0 mmAl filtration. PMID- 10474498 TI - High dose rate brachytherapy practice for the treatment of gynaecological cancers in the UK. AB - A summary of UK high dose rate brachytherapy practice in gynaecological cancer is presented. There appears to be relatively good uniformity in dose prescription and biological effective doses, which represents a considerable improvement from the findings of a previous report of UK low dose rate brachytherapy practice in 1991. Individual details of the dose schedules used at each treatment centre are presented. PMID- 10474499 TI - Estimating the effective dose to children undergoing heart investigations--a phantom study. AB - The aim of the investigation was to assess the conversion factor (F) for derivation of effective dose from measured dose-area product (DAP) during radiological examination of congenital heart diseases. Two anthropomorphic phantoms corresponding to a 1-year-old and a 5-year-old child were irradiated at several projections to imitate irradiation conditions at heart examinations. Organ doses were measured using thermoluminescent dosimeters for calculation of mean organ doses and effective dose according to ICRP. DAP values were measured simultaneously. The conversion factor (F) was calculated from the ratio of effective dose (mSv) to DAP value (Gy cm2). The conversion factor (F) correlated strongly to the size of the phantom but less to the irradiation projection. However, at major beam angulation and at lateral projection F deviated note worthily from that obtained at true or slightly angulated frontal views. Effective dose can therefore be estimated from the DAP values at heart investigation using two different F values. The following values are recommended for PA and lateral view respectively: for children weighing 7-11 kg, 1.8 and 1.4; for children weighing 15-26 kg 0.9 and 0.7. PMID- 10474500 TI - Value of a multileaf prescription preparation system for palliative external beam irradiation. AB - In palliative treatment, irregularly shaped fields are used to reduce side effects and can improve, or avoid, field matching. We investigated the effectiveness of a multileaf collimator (MLC) supported by a digitizing data entry system in the palliative radiotherapy treatment of 66 patients and compared it with conventional shielding with geometrically shaped blocks. After conventional simulation of rectangular fields, irregular field shapes were marked on the simulator film in 17 patients (27%) to reduce radiotherapy related side effects. Individual leading was performed with an MLC. Digitizing and fitting of the optimum leaf position were carried out using a multileaf preparation system (MLP, Elekta, Crawley, UK). Target volumes included bone metastases in the pelvis, spine and extremities, mediastinal soft tissues, lymph nodes and central nervous system. In 10 patients, treated with a parallel pair for pelvic metastases, MLC and conventional shielding were prospectively compared with regard to time requirements and area shielded. Compared with conventional blocking, the mean simulation, preparation and treatment time required for MLP fields was shorter (9.55 +/- 1.44 min vs 16.90 +/- 2.64 min, and 5.50 +/- 1.14 min vs 8.97 +/- 1.75 min). The mean shielded area was 31 cm2 larger for MLC fields compared with geometrically shaped blocks (p < 0.05). Compared with cerrobend blocking, the use of an MLC, supported by preparation data entry software, is more flexible and reduces radiotherapy resources. Therefore, a preparation data entry system as a separate device, or integrated into the treatment planning system, is a useful tool in palliative treatment. PMID- 10474501 TI - The use of independent collimation in the treatment of Graves' ophthalmopathy. AB - Graves' ophthalmopathy is an infiltrative disease of the orbit and randomized trials show that radiotherapy should be the first treatment of choice. This article describes how linear accelerators with independent collimation can be used for setting up purposes, in place of lead blocks, to shield the lens. This benefits the patient and radiographer by being quicker, more accurate and easier to set up. PMID- 10474502 TI - Polypoid endobronchial Hodgkin's disease with pneumomediastinum. AB - We report a 23-year-old man with polypoid endobronchial Hodgkin's disease which presented with intermittent multilobar atelectasis and spontaneous pneumomediastinum. Chest radiograph and CT findings are demonstrated. PMID- 10474503 TI - Right atrial metastatic melanoma detected by dynamic contrast enhanced spiral CT. AB - The use of and transoesophageal echocardiography has been well described in the diagnosis of intracardiac metastases. We present a case of a right atrial mass in a patient with metastatic melanoma which was unexpectedly detected by dynamic enhanced spiral CT. The diagnosis was subsequently confirmed by MRI. It is likely that the increasingly widespread use of spiral CT will result in a greater detection rate of intracardiac tumours. PMID- 10474504 TI - Multiple pyogenic granuloma demonstrated by SPECT using 99Tcm-labelled red blood cells. AB - Pyogenic granuloma is a polypoid form of capillary haemangioma that occurs on the skin and mucosal surface. The eruption of multiple pyogenic granuloma is a rare phenomenon. We report the case of a 73-year-old male with a 2 year history of intermittent lower gastrointestinal bleeding. 99Tcm-labelled red blood cell scintigraphy was performed to rule out the possibility of capillary haemangioma on the mucosal surface of the gastrointestinal tract. The images showed disseminated haemangiomas involving the skull, abdominal wall, intestine, scrotum and right leg. The final diagnosis of multiple pyogenic granuloma was made by skin biopsy. To the best of our knowledge, this is the first case using 99Tcm labelled red blood cell scintigraphy to demonstrate multiple pyogenic granuloma. PMID- 10474505 TI - MRI features of Balo's concentric sclerosis. AB - We report MRI findings in a 56-year-old woman with Balo's concentric sclerosis (BCS) who initially presented with a progressive hemiparesis. MRI showed two lesions with a concentric pattern in the left frontoparietal region and a laminated, arcuate pattern in the right frontal region. These patterns were best seen in post-contrast images and were consistent with BCS. In addition, there were several small cerebral multiple sclerosis-like plaques. The clinical symptoms improved and the MR findings regressed after corticosteroid therapy. The patient had completely recovered 12 months later, except for mild right hand numbness. MRI showed further regression of the lesions, but the concentric pattern was still present. This case demonstrated that BCS can run a benign prolonged course and may persist for a long time. Concentric or laminated contrast enhancement in the acute phase may suggest that bands of demyelination in BCS occur synchronously rather than successively. PMID- 10474506 TI - Pachymeningitis and optic neuritis in rheumatoid arthritis: MRI findings. AB - Rheumatoid arthritis is a systemic disease in which cerebral and eye involvement is neither common nor fully understood. Although it is rarely the cause of pachymeningitis and optic neuritis, rheumatoid arthritis should always be kept in mind in these two conditions. We present a 52-year-old male with an 8 month history of rheumatoid arthritis who was referred to the neurology department with headache and decreasing vision and was diagnosed as having rheumatoid pachymeningitis and optic neuritis on the basis of MRI findings. PMID- 10474507 TI - Dissecting vertebral artery aneurysm: diagnosis and coil embolization. AB - Dissecting aneurysm of the intradural vertebral artery, a rare cause of stroke and sub-arachnoid haemorrhage, is associated with a poor neurological outcome if the disease is unrecognized and remains untreated. Clinical manifestations may be delayed or overlooked and non-invasive radiological investigations demonstrate little or no abnormality. We describe the diagnosis and successful endovascular occlusion of a dissecting vertebral artery aneurysm using detachable microcoils; a novel technique that has advantages over conventional therapeutic procedures. PMID- 10474508 TI - Diffuse retroperitoneal amyloidosis due to renal cell carcinoma. AB - The case of a 61-year-old man with haematuria, in whom CT demonstrated a solid renal mass and extensive retroperitoneal infiltration, is presented. This combination of CT findings resulted in significant diagnostic uncertainty. Pathological analysis demonstrated renal cell carcinoma, with secondary amyloidosis in the retroperitoneum. Amyloidosis secondary to renal cell carcinoma has not been previously described as a cause of retroperitoneal infiltration. PMID- 10474509 TI - Differential diagnosis of gynaecological "stained glass" tumours on MRI. AB - Although multilocular cystic gynaecological masses in which the loculi show variable signal intensity on both T1 and T2 weighted images have been considered to be mucinous cystadenoma or adenocarcinoma, other gynaecological tumours can demonstrate this "stained glass" appearance. These include mature cystic teratoma, fibrothecoma, endometrioma, Brenner's tumour of the ovary and degenerated leiomyoma of the uterus, all of which may mimic mucinous tumours of the ovaries. PMID- 10474510 TI - A case of acute right-sided abdominal pain. PMID- 10474511 TI - The molecular basis of disorders of red cell enzymes. PMID- 10474512 TI - The molecular basis of disorders of the red cell membrane. PMID- 10474513 TI - Human papillomavirus and the development of non-melanoma skin cancer. AB - Human papillomaviruses (HPV) are increasingly recognised as important human carcinogens. The best established association with human malignancy is that of high-risk mucosal HPV types and anogenital cancer. HPV-induced transformation of anogenital epithelia has been the subject of intense research which has identified the cellular tumour suppressor gene products, p53 and pRB, as important targets for the viral oncoproteins E6 and E7 respectively. Certain HPV types are also strongly associated with the development of non-melanoma skin cancer in the inherited disorder epidermodysplasia verruciformis (EV). However, in contrast with anogenital malignancy the oncogenic mechanisms of EV-HPV types remain uncertain, and there appears to be a crucial additional requirement for ultraviolet radiation. Cutaneous HPV types in the general population are predominantly associated with benign viral warts, but a role in non-melanoma skin cancer has recently been postulated. Polymerase chain reaction based HPV detection techniques have shown a high prevalence of HPV DNA, particularly in skin cancers from immunosuppressed patients and to a lesser extent in malignancies from otherwise immunocompetent individuals. No particular HPV type has yet emerged as predominant, and the role of HPV in cutaneous malignancy is unclear at present. It remains to be established whether HPV plays an active or purely a passenger role in the evolution of non-melanoma skin cancer. PMID- 10474514 TI - Supplying commercial biomedical companies from a human tissue bank in an NHS hospital--a view from personal experience. AB - NHS histopathology laboratories are well placed to develop banks of surgically removed surplus human tissues to meet the increasing demands of commercial biomedical companies. The ultimate aim could be national network of non-profit making NHS tissue banks conforming to national minimum ethical, legal, and quality standards which could be monitored by local research ethics committees. The Nuffield report on bioethics provides ethical and legal guidance but we believe that the patient should be fully informed and the consent given explicit. Setting up a tissue bank requires enthusiasm, hard work, and determination as well as coordination between professionals in the NHS trust and in the commercial sector. The rewards are exiting new collaborations with commercial biomedical companies which could help secure our future. PMID- 10474515 TI - Multiplex polymerase chain reaction for the simultaneous detection of Mycoplasma pneumoniae, Chlamydia pneumoniae, and Chlamydia psittaci in respiratory samples. AB - AIMS: To develop a multiplex polymerase chain reaction (PCR) for the simultaneous detection of Mycoplasma pneumoniae, Chlamydia pneumoniae, and Chlamydia psittaci in respiratory samples. METHODS: Oligonucleotide primers for the amplification of the DNA of these three organisms were optimised for use in combination in the same reaction. PCR products were detected by hybridisation with pooled internal probes using an enzyme linked immunosorbent assay. Those with positive signals were further differentiated using species specific probes. Quality of DNA extraction and PCR inhibition were controlled by amplification of a human mitochondrial gene. A panel of 53 respiratory samples with known results was evaluated blindly. This was followed by a retrospective study on sputa collected from 244 patients with suspected community acquired pneumonia. RESULTS: The multiplex assay had a lower sensitivity than PCR with individual primers by about one log. The resultant sensitivity was considered acceptable for diagnostic use. Of the panel of 53 samples, nine of 11 M pneumoniae, 11 of 11 C pneumoniae, six of seven C psittaci, and 24 of 24 negative samples were correctly identified. Of the 244 patients with pneumonia, seven (2.9%) had detectable M pneumoniae, six (2.5%) had C pneumoniae, and one (0.4%) had C psittaci. The case notes from 11 patients were studied. The PCR finding was of possible significance in at least eight of these patients. CONCLUSIONS: This multiplex PCR assay has the potential to be used as a diagnostic and epidemiological tool. Further prospective studies are needed to establish its clinical value. PMID- 10474516 TI - Group C streptococci isolated from throat swabs: a laboratory and clinical study. AB - AIMS: To determine the prevalence of beta haemolytic, Lancefield group C streptococci in throat swabs taken in routine clinical practice, and correlate the species identified with presenting clinical features. METHODS: One year, laboratory based prospective study, using a questionnaire to elicit clinical information. RESULTS: 4.4% of throat swabs yielded group C streptococci, of which 38% belonged to S equisimilis and 53% to S anginosus-milleri group (SAM). Pyrexia was more common in patients with S equisimilis, but other clinical features did not differ significantly between the two groups. No S zooepidemicus was isolated. CONCLUSIONS: Species identification of group C streptococci from throat swabs does not appear to be clinically useful in this patient population. However, the prevalence and spectrum of organisms is similar to that reported in N America, where studies suggest a possible role in some cases of severe pharyngitis. Observational studies such as this lack power to resolve the issue of pathogenicity, for which a placebo controlled trial of antibiotic treatment is ideally required. PMID- 10474517 TI - Prevalence of HIV and hepatitis C markers among a cadaver population in Milan. AB - AIMS: To examine the distribution of serological markers for HIV and hepatitis C virus (HCV) infection in a medicolegal population; to compare prevalence of HIV and HCV markers and cause of death; and to evaluate the risk of potential infection to personnel involved in medicolegal incidents and procedures. METHODS: Blood samples were collected at necropsy from 328 males and 69 females, aged 16 to 50 years at time of death, and tested for antibodies to HIV and HCV. The individuals were classified according to cause of death and whether there was known antemortem risk of infection. RESULTS: Overall, 134 subjects gave positive test results: 20 for anti-HIV, 69 for anti-HCV, and 45 for both. By cause of death, the total number of positives (and negatives) with the pairs of figures referring, respectively, to patients with and without known antemortem risks were: natural causes 3 (1), 8 (32); AIDS 2 (0), 0 (0); homicide 0 (2), 5 (24); suicide 3 (0), 9 (69); road traffic accidents 1 (0), 9 (81); other accidents 2 (3), 1 (12); drug overdose 74 (17), 7 (9); unknown causes 3 (2), 7 (11). CONCLUSIONS: The cases tested represented a predominantly young male population with a high prevalence of serological markers for HIV and HCV infection. The distribution of HIV and HCV positivity varied with the cause of death, probably reflecting the known association between high risk behaviour and infection. However, a substantial number of cases with no known risks also had markers for HIV and HCV, suggesting that there is a large unrecognised pool of potential infection in medicolegal practice. PMID- 10474518 TI - Comparative study of seven commercial yeast identification systems. AB - AIMS: To compare the performance of seven commercial yeast identification methods with that of a reference method, and to compare the costs of the commercial kits. METHODS: Clinical yeast isolates (n = 52), comprising 19 species, were identified using Vitek, Api ID 32C, Api 20C AUX, Yeast Star, Auxacolor, RapID Yeast Plus system, and Api Candida and compared with a reference method which employed conventional tests. RESULTS: The percentage of correctly identified isolates varied between 59.6% and 80.8%. Overall, the highest performance was obtained with Api Candida (78.8%) and Auxacolor (80.8%). Among germ tube negative yeast isolates, Auxacolor and Api Candida both identified 93.1% of isolates correctly. All systems failed to identify C norvegensis, C catenulata, C haemulonii, and C dubliniensis. In comparison with Auxacolor, the Api Candida is less expensive and requires less bench time. CONCLUSIONS: Auxacolor and Api Candida appeared to be the most useful systems for identification of germ tube negative yeast isolates in clinical microbiology laboratories, although one should be aware that several germ tube negative Candida species cannot be identified by these systems. PMID- 10474519 TI - IgA anti-tissue transglutaminase as a diagnostic marker of gluten sensitive enteropathy. AB - AIMS: To compare and contrast the sensitivity, specificity, and positive predictive values of IgA antibodies to guinea pig tissue transglutaminase (ELISA), endomysium, and reticulin (immunofluorescence), and gliadin (ELISA), and IgG antibodies to gliadin and tissue transglutaminase. METHODS: Sera from 27 newly diagnosed patients with coeliac disease, 65 biopsied gastrointestinal disease controls, and 50 consecutive blood donors were tested. All cases were adults. RESULTS: IgA anti-tissue transglutaminase showed a sensitivity of 85% (23/27 coeliac disease cases seropositive), specificity 97% (2/65 controls and one blood donor showing low titre positivity), and a positive predictive value of 92%. High titre anti-tissue transglutaminase was only seen in coeliac disease. Disease controls with mucosal damage unrelated to gluten enteropathy were IgA anti-tissue transglutaminase negative. Sensitivity, specificity, and positive predictive values for IgA anti-endomysial antibody (monkey oesophagus) were 100%, 100%, and 100%, respectively, and for IgA anti-gliadin, 93%, 95%, and 89%, respectively. CONCLUSIONS: Tissue transglutaminase is a major autoantigen in coeliac disease. IgA (but not IgG) anti-tissue transglutaminase, especially when in high titre, is closely associated with coeliac disease, but low titres may not be disease specific. In this small pilot study, the established IgA anti endomysial assay was the superior test. PMID- 10474520 TI - Endometrial pathology in postmenopausal tamoxifen treatment: comparison between gynaecologically symptomatic and asymptomatic breast cancer patients. AB - AIMS: To evaluate whether endometrial pathology is more likely to be diagnosed in gynaecologically symptomatic rather than in gynaecologically asymptomatic postmenopausal breast cancer patients with tamoxifen treatment; and to evaluate the possible influence of various clinical factors on the incidence of endometrial pathology. METHODS: Endometrial histological findings, transvaginal ultrasonographic endometrial thickness, demographic characteristics, health habits, and risk factors for endometrial cancer were compared between 14 gynaecologically symptomatic (group I) and 224 gynaecologically asymptomatic (group II) postmenopausal breast cancer patients with tamoxifen treatment. RESULTS: Overall, 28.6% of the study population had endometrial pathology. The incidence of overall positive endometrial histological findings was significantly higher in group I than in group II (92.9% v 24.6%, p < 0.0001). Atrophic endometrium was more common in group II than in group I (75.3% v 7.1%, p < 0.0001). Most other endometrial pathology was significantly more common in group I than in group II (endometrial hyperplasia, 35.7% v 5.6%, p < 0.0001; endometrial polyps, 35.7% v 13.4%, p < 0.0111; endometrial carcinoma, 21.5% v 0.9%, p < 0.0001). Endometrial pathology appeared considerably later in the gynaecologically asymptomatic patients than in gynaecologically symptomatic patients (p = 0.0002). Vaginal bleeding or spotting occurred exclusively in group I. The incidence of endometrial pathology in the entire study population was consistent with that reported elsewhere, and higher than that reported for healthy postmenopausal women. CONCLUSIONS: Endometrial pathology is more likely to be diagnosed in gynaecologically symptomatic postmenopausal breast cancer patients with tamoxifen treatment, and after a shorter duration of time, than in gynaecologically asymptomatic patients. PMID- 10474521 TI - Immunohistochemical differentiation between primary adenocarcinomas of the ovary and ovarian metastases of colonic and breast origin. Comparison between a statistical and an intuitive approach. AB - AIM: To discriminate between adenocarcinomas that are primary to the ovary and metastatic to the ovary, especially of colonic and breast origin, by immunohistochemistry, using stepwise discriminant analysis or a decision tree. METHODS: 312 routinely processed, formalin fixed tissue specimens were used. The tumours were divided into a learning set (n = 159), composed of primary tumours of ovary, breast, and colon, and a test set, comprising 134 metastases from these sites and an additional 19 primary ovarian carcinomas. The immunohistochemical panel was composed of antibodies against cytokeratin 7 (CK7) and 20 (CK20), CA125, vimentin, carcinoembryonic antigen (CEA), gross cystic disease fluid protein-15 (GCDFP-15), and the oestrogen receptor (ER). The staining results of the tumours were expressed as the product of the staining intensity and the percentage of positive tumour cells. Analyses were first performed on the learning set and then evaluated on the test set. RESULTS: Although the immunostaining patterns showed a considerable overlap between the three types of adenocarcinoma, the breast carcinomas were typically positive for GCDFP-15 and often for ER, and negative for vimentin. Ovarian carcinomas were always positive for CK7 and to a lesser extent for CA125. Colonic carcinomas showed prominent positivity for CEA and CK20, while no staining was seen for ER and vimentin. In discriminant analysis, six antibodies (alpha CK7, alpha CK20, alpha CA125, alpha CEA, alpha ER, and alpha GCDFP-15) appeared to be necessary for optimal classification: 89% of the learning set and 82% of the test set were classified correctly. In the decision tree, only four antibodies (alpha CK7, alpha CEA, alpha ER, and alpha GCDFP-15) were used to obtain a correct classification score of 89% for the learning set and 84% for the test set. CONCLUSIONS: Using a semiquantitative assessment of the immunostaining results by a restricted panel of six antibodies with stepwise discriminant analysis, 80-90% of the adenocarcinomas of colon, breast, and ovary can be correctly classified. Discriminant analysis is computer aided and therefore an easy method and for each case a probability value of the classification result is obtained. The intuitive decision tree method provides a slightly better result, requires only four antibodies, and offers a more practical method for the surgical pathologist. PMID- 10474522 TI - Mutations of p53 gene and SV40 sequences in asbestos associated and non-asbestos associated mesotheliomas. AB - AIM: To examine mesotheliomas for a possible relation between p53 immunostaining, p53 gene mutation, simian virus 40 (SV40), and asbestos exposure. METHODS: Paraffin sections from 11 mesotheliomas were used for p53 immunostaining and also to extract DNA. This was analysed for the presence of mutations in exons 5 to 8 of the p53 gene using a "cold" single strand conformational polymorphism method, together with sequencing. The DNA from the paraffin sections was also used to search for SV40 sequences. A 105 base pair segment at the 3' of the SV40 large T antigen (Tag) was targeted and any PCR amplification products were sequenced to confirm that they were of SV40 origin. EDAX electron microscopic differential mineral fibre counts were performed on dried lung tissue at a specialist referral centre. RESULTS: The fibre counts showed that seven of the mesotheliomas were associated with abnormally high asbestos exposure. Of these, two showed p53 immunostaining, none showed p53 gene mutation, and five showed SV40. Of the four other mesotheliomas, three showed p53 immunostaining, one showed a (silent) p53 mutation, and none showed SV40. The difference in frequency of SV40 detection was significant at the p < 0.05 level. CONCLUSIONS: Immunostaining for the p53 gene was relatively common but p53 mutations were rare in this series. SV40 virus sequence was detected in five of seven asbestos associated mesotheliomas but in none of the non-asbestos-associated mesotheliomas. This suggests there may be a synergistic interaction between asbestos and SV40 in human mesotheliomas. A study with a larger number of cases is needed to investigate these observations further. PMID- 10474523 TI - Focal lymphoid aggregates (nodules) in bone marrow biopsies: differentiation between benign hyperplasia and malignant lymphoma--a practical guideline. AB - AIMS: To provide practical guidelines for the differentiation between benign and malignant focal lymphoid aggregates (lymphoid nodules) in routinely referred bone marrow trephine biopsies, using a synoptic approach including clinical data and histological workup. METHODS: For easy identification of very small lymphoid infiltrates the chloroacetate esterase stain was applied as a screening procedure. This allowed the identification of 491 formalin fixed, paraffin wax embedded specimens with one or more lymphoid nodules. Examination of lymphoid infiltrates included such variables as histotopography, demarcation, cytology, reticulin fibres, and immunohistochemistry with a set of monoclonal antibodies (CD20, CD45R, CD45R0, CD3, CD43). Evaluation of clinical and morphological data was carried out independently. In case of malignant lymphomas, a correlation with corresponding lymph node findings was made. RESULTS: 352 patients had benign focal lymphoid aggregates usually associated with systemic autoimmune diseases, chronic myeloproliferative disorders, toxic myelopathy, and viral infections. Discrete nodular infiltrates of (small cell) malignant lymphomas (n = 93) simulating benign hyperplasia were found in chronic lymphocytic leukaemia, germinal centre cell lymphomas (CB-CC), and lymphoplasmacytic/cytoid lymphomas (LPI). In addition to immunoreactivity, certain histological variables proved distinctive. These were: (1) histotopography, that is, localisation of the lymphoid aggregates within the bone marrow space; (2) relation to the surrounding tissue: margination or interstitial spillage of lymphoid cells; and (3) increase in reticulin fibres. CONCLUSIONS: A combined diagnostic procedure identifying several distinctive features, in particular histotopography and immunohistochemistry, provides a most promising way of discriminating reactive from neoplastic lymphoid nodules in the bone marrow. PMID- 10474524 TI - An unusual presentation of systemic mastocytosis. AB - A 47 year old man presented with mastocytosis, a disease process characterised by proliferation of mast cells. The clinical features and outcome are discussed. PMID- 10474525 TI - Lymphocytic venulitis: an unusual association with microscopic colitis. AB - A 79 year old man presented with occult gastrointestinal bleeds and anaemia for two years. He had received 40 units of blood over a period of one year, following which he had a subtotal colectomy as no definite cause of the bleeding was apparent. Macroscopically the colon appeared unremarkable. Light microscopy showed prominent lymphocytic venulitis in the proximal portion, gradually merging into lymphocytic and collagenous colitis distally. PMID- 10474526 TI - Phaeochromocytoma of the spermatic cord. AB - Phaeochromocytoma of the spermatic cord is very rare. It can arise anywhere in the distribution of cells of neural crest origin, but 80-90% arise in the adrenal medulla and only 3% are extra-abdominal. A small tumour may be asymptomatic because insufficient catecholamines are secreted to cause haemodynamic disturbance. PMID- 10474527 TI - Intraglomerular metastasis: a necropsy study. AB - Intraglomerular metastasis is a rare phenomenon. Four cases are reported here. The primary malignancies in these four cases were squamous cell carcinoma of the lung, adenocarcinoma of pancreas, haematological malignancy with a malignant intrapulmonary teratoma, and undifferentiated pleural mesothelioma. There was no significant renal functional impairment in any case except for mild proteinuria in one. Histopathology showed tumour cells in the mesangium, in the glomerular tufts replacing the endothelial cells, as well as in an extracapillary location replacing the parietal cells. In one case, tumour cells formed crescent-like structures. Such tumour deposits were better appreciated with special stains. The presence of intraglomerular metastasis indicates dissemination of the malignancy and hence a poor prognosis even though there is no significant impairment of renal function. PMID- 10474528 TI - Adenocarcinoma arising within a tailgut cyst: clinicopathological description and follow up of an unusual case. AB - Full clinicopathological details and clinical follow up of a case of malignant transformation within a tailgut cyst are presented. A 43 year old woman presented with signs and symptoms of an imminent threatened abortion. Routine examination identified a coincidental, asymptomatic retrorectal/presacral mass. Following imaging studies, surgical resection was carried out and an adenocarcinoma arising within a pre-existent tailgut cyst was identified by microscopy. Four years later the patient presented with neurological symptoms consistent with local recurrence of the tumour. Surgical biopsies confirmed this diagnosis and she was subsequently started on chemotherapy. She died soon after from a cause unrelated to the disease, after declining further active intervention. Differential diagnosis of such cases includes (cystic) teratoma, epidermal cyst, rectal duplication cyst, anal gland cyst and carcinoma, extension of local carcinoma, and metastatic disease. It is recommended that these lesions be completely excised when detected incidentally. PMID- 10474529 TI - Coexistent gastric MALT lymphoma and Kaposi sarcoma in an HIV positive patient. AB - A 47 year old HIV positive male presented with haematemesis and epigastric pain. A gastrectomy was performed for intractable bleeding. The cause of the haematemesis proved to be a Kaposi sarcoma of the stomach which had resulted in mucosal ulceration. Several other smaller foci of Kaposi sarcoma were also present. Coexistent with the Kaposi sarcoma was a dense lymphoid infiltrate with lymphoid follicles and reactive germinal centres. Centrocyte-like cells caused marked effacement and destruction of gastric glands with the formation of lymphoepithelial lesions, typical of a MALT lymphoma. These cells were of B cell lineage and some expressed the HIV antigen, p24. Follicular dendritic cells and macrophages within germinal centres were also p24 positive. Immunohistochemistry and in situ hybridisation did not detect Epstein-Barr virus. Although Helicobacter pylori was not identified by light microscopy in the sections sampled, this does not preclude its possible role, with other cofactors such as HIV, in the causation of the MALT lymphoma. PMID- 10474530 TI - Rights of possession in human corpses. PMID- 10474531 TI - Evaluation research in occupational health services: general principles and a systematic review of empirical studies. AB - OBJECTIVES: To study the nature and extent of evaluation research in occupational health services (OHSs). METHODS: Literature review of evaluation research in OHSs. On the basis of a conceptual model of OHS evaluation, empirical studies are categorised into aspects of input, process, output, outcome, and OHS core activities. RESULTS: Many methods to evaluate OHSs or OHS activities exist, depending on the objective and object of evaluation. The amount of empirical studies on evaluation of OHSs or OHS activities that met the non-restrictive inclusion criteria, was remarkably limited. Most of the 52 studies were more descriptive than evaluative. The methodological quality of most studies was not high. A differentiated picture of the evidence of effectiveness of OHSs arises. Occupational health consultations and occupational rehabilitation are hardly studied despite much time spent on the consultation by occupational physicians in most countries. The lack of effectiveness and efficiency of the pre-employment examination should lead to its abandonment as a means of selection of personnel by OHSs. Periodic health monitoring or surveillance, and education on occupational health hazards can be carried out with reasonable process quality. Identification and evaluation of occupational health hazards by a workplace survey can be done with a high output quality, which, however, does not guarantee a favourable outcome. CONCLUSIONS: Although rigorous study designs are not always applicable or feasible in daily practice, much more effort should be directed at the scientific evaluation of OHSs and OHS instruments. To develop evidence-based occupational health care the quality of evaluation studies should be improved. In particular, process and outcome of consultation and rehabilitation activities of occupational physicians need to be studied more. PMID- 10474532 TI - Evaluation of exposure to ethylene glycol monoethyl ether acetates and their possible haematological effects on shipyard painters. AB - OBJECTIVES: To evaluate exposure to mixed solvents containing ethylene glycol monoethyl ether acetate (EGEEA) in shipyard painters, to determine if EGEEA is toxic to the bone marrow. METHODS: An industrial hygiene survey was performed to identify exposure to EGEEA of two groups of shipyard painters, a low exposure group (n = 30) and a high exposure group (n = 27). Urinary ethoxyacetic acid and methyl hippuric acid as well as haemoglobin, packed cell volume, red cell indices, total and differential white blood cell counts (WBCs), and platelet count for the shipyard painters and the control subjects were measured. RESULTS: The mean (range) exposure concentration (ppm) to EGEEA in the high and low exposure groups were 3.03 (not detectable to 18.27), 1.76 (not detectable to 8.12), respectively. The concentrations of methyl hippuric acid and ethoxyacetic acid in the high exposure group were significantly higher than those in the control group. The mean WBCs in the high exposure group were significantly lower than in the control group, and a significant proportion, six (11%) of the 57 painters, were leucopenic; none of the controls were affected. CONCLUSION: The high rate of possible haematological effects among shipyard painters and a hygienic evaluation of their working environment in the present study suggests that EGEEA might be toxic to bone marrow. PMID- 10474534 TI - Neuropsychological symptoms in Chinese male and female painters: an epidemiological study in dockyard workers. AB - OBJECTIVES: To study the prevalences of neuropsychological symptoms in male and female dockyard painters in China and to compare them with those in British dockyard painters. METHODS: All 116 painters were identified, active and retired, who had been employed in two Chinese dockyards for at least 1 year together with a matched random sample of 263 dockyard non-painters. Neuropsychological and personality questionnaires that we had used previously in a study of United Kingdom dockyard painters were used, translated into Chinese. Neuropsychological symptoms in painters and controls were compared, adjusting for age, educational level, smoking, alcohol intake, and personality. RESULTS: The response rate was 94% for painters and 97% for controls. Highly significant excesses of symptoms were found in painters, suggestive of neuropsychological dysfunction. Both male and female painters showed higher relative risks than were found in similar tradespeople in the United Kingdom. The relative risk increased with increasing score of both neurological and psychological symptoms. Relative risk of having a high symptom score, compared with controls and adjusted for confounders, was 6.61 (95% confidence interval (95% CI) 2.36 to 18.50) for 2-15 years exposure, 14.88 (5.74 to 38.56) for 16-22 years and 9.42 (3.97 to 22.36) for > or = 22 years. CONCLUSION: These results indicate that neuropsychological symptoms are associated with heavy exposure to painting work in China, and that the phenomenon is likely to be found worldwide wherever there is such exposure to solvent based paints. The high response rate in this study answers a possible criticism of the earlier United Kingdom study. PMID- 10474533 TI - Health effects of solvent exposure among dockyard painters: mortality and neuropsychological symptoms. AB - OBJECTIVES: To study mortality and prevalence of neuropsychological symptoms among a cohort of painters known to have been heavily exposed to organic solvents. METHODS: A mortality study of 1292 male painters who had worked in a dockyard in Scotland for > or = 1 year between 1950 and 1992 comprised a nested cross sectional study of 953 surviving painters from the cohort and 953 male non painters randomly selected from the local population and a case-control study of those with high symptom scores. Mortality, symptoms, and risks associated with painting, adjusting for age, education, smoking, alcohol, and personality were measured. RESULTS: The proportional mortality ratio for all cancers was not increased significantly (110 (95% confidence interval (95% CI) 84 to 143), except for a possible excess of deaths from ischaemic heart disease (132, 105 to 164). Standardised mortality ratios were not significantly increased. Among the 260 surviving painters and 539 community controls who responded to the questionnaire there was a significant excess of symptoms among painters; adjusted relative risk (RR) increased significantly with increasing symptom score. These RRs suggested an exposure-response relation; for a high score (12-22) for all symptoms RR was 2.27 (1.20 to 4.30) for 1-4 years of exposure, 2.42 (1.18 to 4.95) for 5-9 years, 2.89 (1.42 to 5.88) for 10-14 years, and 3.41 (1.82 to 6.36) for 15-41 years, compared with controls. In multivariate analyses, painting exposure, and aging were associated with high symptom scores and there was again an increased risk relative to time worked as a painter. CONCLUSION: This study supports the hypothesis that heavy and prolonged exposure to paint solvents leads to neuropsychological ill health. PMID- 10474535 TI - Long-term respiratory health effects of the herbicide, paraquat, among workers in the Western Cape. AB - OBJECTIVE: To evaluate the possible effects of paraquat spraying among workers on deciduous fruit farms in the Western Cape, South Africa. Paraquat is a commonly used herbicide world wide and is a well documented cause of pulmonary fibrosis in studies of laboratory animals and in humans after exposure to a high dose (usually accidental or as parasuicide). The respiratory effects of long term, low dose exposure to paraquat have not been fully evaluated. METHODS: A cross sectional study of 126 workers. Administered questionnaires generated information on exposure, respiratory symptoms, and potential confounding variables. Spirometry and gas transfer were measured and chest radiographs performed. Oxygen desaturation on exercise testing was by oximetry during a modified stage one exercise test. RESULTS: No association was found between long term exposure to paraquat and reported symptoms, spirometry (forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), FEV1/FVC) and gas transfer (TLCO and KCO) or chest radiography. Multivariate analysis showed a significant relation between measures of long term exposure to paraquat and arterial oxygen desaturation during exercise independent of short term exposure. CONCLUSION: Previous studies have not shown a significant relation between measures of exposure to paraquat and standard tests of lung function. Arterial oxygen desaturation during exercise represents a more sensitive test. The findings indicate that working with paraquat under usual field conditions is associated with abnormal exercise physiology in a dose dependent fashion independent of recent exposure and acute poisoning events. PMID- 10474536 TI - Germicidal ultraviolet irradiation in air conditioning systems: effect on office worker health and wellbeing: a pilot study. AB - OBJECTIVES: The indoor environment of modern office buildings represents a new ecosystem that has been created totally by humans. Bacteria and fungi may contaminate this indoor environment, including the ventilation systems themselves, which in turn may result in adverse health effects. The objectives of this study were to test whether installation and operation of germicidal ultraviolet (GUV) lights in central ventilation systems would be feasible, without adverse effects, undetected by building occupants, and effective in eliminating microbial contamination. METHODS: GUV lights were installed in the ventilation systems serving three floors of an office building, and were turned on and off during a total of four alternating 3 week blocks. Workers reported their environmental satisfaction, symptoms, as well as sickness absence, without knowledge of whether GUV lights were on or off. The indoor environment was measured in detail including airborne and surface bacteria and fungi. RESULTS: Airborne bacteria and fungi were not significantly different whether GUV lights were on or off, but were virtually eliminated from the surfaces of the ventilation system after 3 weeks of operation of GUV light. Of the other environmental variables measured, only total airborne particulates were significantly different under the two experimental conditions--higher with GUV lights on than off. Of 113 eligible workers, 104 (87%) participated; their environmental satisfaction ratings were not different whether GUV lights were on or off. Headache, difficulty concentrating, and eye irritation occurred less often with GUV lights on whereas skin rash or irritation was more common. Overall, the average number of work related symptoms reported was 1.1 with GUV lights off compared with 0.9 with GUV lights on. CONCLUSION: Installation and operation of GUV lights in central heating, ventilation and air conditioning systems of office buildings is feasible, cannot be detected by workers, and does not seem to result in any adverse effects. PMID- 10474537 TI - Follow up investigation of workers in synthetic fibre plants with humidifier disease and work related asthma. AB - OBJECTIVE: To investigate the clinical and sociomedical outcome in patients with various clinical manifestations of humidifier disease and work related asthma after removal from further exposure. METHODS: Follow up investigation (range 1-13 years) of respiratory symptoms, spirometry, airway responsiveness, sickness absence, and working situation in patients with (I) humidifier fever (n = 12), (II) obstructive type of humidifier lung (n = 8), (III) restrictive type of humidifier lung (n = 4), and (IV) work related asthma (n = 22). All patients were working at departments in synthetic fibre plants with microbiological exposure from contaminated humidification systems or exposure to small particles (< 1 micron) of oil mist. RESULTS: At follow up patients with work related asthma were less often symptom free (37%, 7/19) than patients with humidifier disease (I, II, III) (67%, 16/24). Mean forced expiratory volume in one second (FEV1) of patients with obstructive impairment had been increased significantly at follow up but still remained below the predicted value. Mean forced vital capacity (FVC) of patients with initially restrictive impairment had returned to normal values at follow up. Airway hyperresponsiveness at diagnosis persisted in patients with obstructive impairment (II + IV 14/17, but disappeared in patients with humidifier fever (3/3) and restrictive type of humidifier lung (2/2). In patients with obstructive impairment (II + IV), FVC and FEV1 at diagnosis were negatively associated with the duration between onset of symptoms and diagnosis and the number of years of exposure. Those with positive pre-employment history of respiratory disease had a lower FEV1 at diagnosis. Sickness absence due to respiratory symptoms decreased in all groups of patients after removal from further exposure, but this was most impressive in patients with the humidifier lung (II, III) and patients with work related asthma (IV). At follow up 83% of the patients were still at work at the same production site, whereas 11% received a disability pension because of respiratory disease. CONCLUSION: In patients with work related respiratory disease caused by exposure from contaminated humidification systems or oil mist, removal from further exposure resulted in clinical improvement, although, especially in those with obstructive impairment, signs persisted. Because of the possibility of transferring patients to exposure free departments most patients could be kept at work. PMID- 10474538 TI - Work related risk factors for musculoskeletal complaints in the spinning industry in Lithuania. AB - OBJECTIVES: To describe the prevalence of self reported musculoskeletal complaints in the back, arms or neck, and legs among workers in the spinning industry, and to investigate the relations between these complaints and work related variables. METHODS: An interview based questionnaire survey was carried out in two spinning industry factories in Lithuania. RESULTS: The study group consisted of all workers in production (n = 363). Symptoms of the legs were the musculoskeletal symptom reported most often (61%). Many subjects had arms or neck (55%) or back problems (28%). 20% had experienced pain from all three sites. Almost 25% had had musculoskeletal pain every day and 16% had experienced constant pain during previous year. Packers had the highest risk of arms or neck problems whereas spinners had the highest risk of back or leg problems. Working in a strained posture (bending, work with arms raised up above shoulder level, and repetitive movements of the fingers) was associated with all three complaints. Only arms or neck complaints were associated with age. CONCLUSIONS: Musculoskeletal disorders are a common problem among workers producing gobelin or synthetic thread in Lithuania and working in a strained posture is a risk factor for developing musculoskeletal disorders in three body sites: legs, arms or neck, and back. To better understand the different aspects of physical load as risk factors, a more detailed study of the frequency of postural changes as well as an observation of individually adopted postures would be necessary. This applies to intervention studies in factories of the spinning industry to prevent complaints of the legs and shoulders. PMID- 10474539 TI - Prevalence and incidence of carpal tunnel syndrome in a meat packing plant. AB - OBJECTIVES: To determine prevalence and incidence of carpal tunnel syndrome (CTS) in a modern meat packing plant. The secondary objective was to explore the relation between ethnicity and CTS. METHODS: Six hundred and sixty five workers were interviewed and examined to find the prevalence of CTS. Subsequently, 421 workers without CTS were followed up and examined at a median interval of 253 days; of those, 333 remained without CTS and were again examined at a median interval of 148 days. RESULTS: The prevalence and incidence of CTS was 21% and 11/100 person-years, respectively. The incidence for Asian mixed, white, and other ethnicities was 12.0, 12.2, and 7.2 cases/100 person-years, respectively. The observed incidence for men and women was 9.7 and 18.4 cases/100 person-years, respectively. This difference was not quite significant (p = 0.068) with an estimated relative risk (women v men) of 1.9 (95% confidence interval (95% CI) 0.9 to 3.8). The interaction between sex and use of tools was significant (p = 0.04), however, although the relative risk for CTS in women who used tools was 4.2 the numbers were small and not significant. The relative risk for men who used tools was 0.64 and not significant. The percentage of incident cases with comorbid disease was only 6.3% (3/47). CONCLUSIONS: The prevalence and incidence of CTS in this workforce were higher than in the general population. However, the prevalence of CTS in this modern, mechanised plant was not significantly different from that reported in older plants. No relation was found between ethnicity, age, body mass index, and CTS for either prevalence or incidence. Comorbid disease among the cases of CTS is significantly less than that found in other industry. PMID- 10474540 TI - Fungal corneal ulcers of onion harvesters in southern Taiwan. AB - Fungal corneal ulcers related to agriculture has been reported throughout the world, especially in tropical areas. Most of them were sporadic and had histories of ocular trauma or use of topical corticosteroids and topical antibiotics. Five onion harvesters had fungal corneal ulcers during the same harvest period in Southern Taiwan. The authors think that this is the first report of a group occurrence relating to agricultural workers. Although all of the patients improved after medical and surgical management, their vision was greatly decreased. It is suggested that the tropical climate, the harvest procedure, the characteristic monsoon, and lack of eye protection were involved. Therefore, the importance of the eye protection, hygiene education, and improving medical care to reduce the occurrence of fungal corneal ulcer in agriculture workers must be emphasised. PMID- 10474541 TI - Suspected nasopharyngeal carcinoma in three workers with long-term exposure to sulphuric acid vapour. AB - Sulphuric acid vapour has been suspected of being an industrial carcinogen. In this study, a cluster is presented of three patients with nasopharyngeal carcinoma (NPC) who worked in the same building of a telecommunications conveyance station in southern Taiwan with long term exposure to sulphuric acid vapour concentrations as high as 0.18 mg/m3. All three workers were diagnosed with NPC within a 5 month period between September 1992, and March 1993. Compared with 19 other healthy workers from the same building, these three workers with NPC had worked significantly longer in this building than had the others (mean (SD) (years): 12.7 (0.6) v 7.4 (4.4); p = 0.01). With an in situ nucleic acid hybridisation and immunostaining method for colocalised Epstein-Barr virus (EBV) and secretory component (SC) protein among biopsy specimens of these three patients with NPCs, it was found that some tumour cells did not contain EBV and SC protein staining signals. These results indicate that EBV infection is not the only risk factor for NPC and long term exposure to relatively low concentrations of sulphuric acid vapour may be associated with the development of NPC. PMID- 10474542 TI - Engine reconditioning workshops: lead contamination and the potential risk for workers: a pilot study. AB - Lead concentrations were measured in surface dust, airborne dust, air, and grinding material from five engine reconditioning workshops to evaluate the impact on blood lead concentrations (PbB) of 10 employees. Lead in the environmental samples ranged from trace amounts to extremely high concentrations (4667 mg/m2). The highest concentrations in surface wipes were found in areas where engine deposits are removed from valves and valve seats. The amounts of lead in long term dustfall accumulation and static air filter samples varied with the position in the workshop and the amount of ventilation. In all but one workshop, the air lead concentrations exceeded Australian occupational guidelines of 150 micrograms Pb/m3. PbB ranged from 4.5 to 25.3 micrograms/dl. There was an empirical relation between the cleanliness, work practices, ventilation of the workshops, lead concentrations in environmental samples and PbB. Office employees not directly exposed to the leaded dust had the lowest PbB. Those who smoked had the highest PbB. Several relatively inexpensive recommendations were made to the owners to minimise exposure of the workers and in most cases these have been implemented. PMID- 10474543 TI - Mind matters: the importance of mental disorders in public health's 21st century mission. PMID- 10474544 TI - A world behaving badly: the global challenge for behavioral surveillance. PMID- 10474545 TI - The global burden of mental disorders. AB - Recent data on the burden of mental disorders worldwide demonstrates a major public health problem that affects patients, society, and nations as a whole. Research must be done to find effective ways to deal with the increasing burden of mental disorders. Given the growing evidence that mental disorders are disorders of the brain and that they can be treated effectively with both psychosocial counseling and psychotropic medications, intervention packages could be developed to deal with the increasing burden. Such packages should be tested for real-world effectiveness and their cost-effectiveness should be demonstrated to guide policymakers to choose from among many other non-mental health interventions. The transportability and sustainability of intervention packages should be studied in public health research and a link between efficacy, effectiveness, cost-effectiveness, generalizability, and sustainability should be demonstrated. The World Health Organization's initiative on the World Mental Health 2000 Survey will provide the first basic epidemiologic data. Together with other data, the initiative will provide solid evidence for including mental disorders into essential treatment packages. In this way, parity can be achieved for mental disorders and mental health can be mainstreamed into health and public health practice. PMID- 10474546 TI - Labor Day and the war on workers. AB - We celebrate Labor Day every year with barbecues and picnics, rarely remembering that the holiday was born in the midst of tremendous labor struggles to improve working conditions. In the last century, 16-hour workdays and 6- and 7-day workweeks led to terribly high injury rates in the nation's mines and mills. Thousands upon thousands of workers died, caught in the grinding machinery of our growing industries. Today, despite improvements, thousands of workers still die in what has been described as a form of war on the American workforce. This commentary reminds us of the historical toll in lives and limbs that workers have paid to provide us with our modern prosperity. It also reminds us that the continuing toll is far too high and that workers who died and continue to die in order to produce our wealth deserve to be remembered and honored on this national holiday. PMID- 10474547 TI - Evaluating the public health impact of health promotion interventions: the RE-AIM framework. AB - Progress in public health and community-based interventions has been hampered by the lack of a comprehensive evaluation framework appropriate to such programs. Multilevel interventions that incorporate policy, environmental, and individual components should be evaluated with measurements suited to their settings, goals, and purpose. In this commentary, the authors propose a model (termed the RE-AIM model) for evaluating public health interventions that assesses 5 dimensions: reach, efficacy, adoption, implementation, and maintenance. These dimensions occur at multiple levels (e.g., individual, clinic or organization, community) and interact to determine the public health or population-based impact of a program or policy. The authors discuss issues in evaluating each of these dimensions and combining them to determine overall public health impact. Failure to adequately evaluate programs on all 5 dimensions can lead to a waste of resources, discontinuities between stages of research, and failure to improve public health to the limits of our capacity. The authors summarize strengths and limitations of the RE-AIM model and recommend areas for future research and application. PMID- 10474548 TI - Public conceptions of mental illness: labels, causes, dangerousness, and social distance. AB - OBJECTIVES: The authors used nationwide survey data to characterize current public conceptions related to recognition of mental illness and perceived causes, dangerousness, and desired social distance. METHODS: Data were derived from a vignette experiment included in the 1996 General Social Survey. Respondents (n = 1444) were randomly assigned to 1 of 5 vignette conditions. Four vignettes described psychiatric disorders meeting diagnostic criteria, and the fifth depicted a "troubled person" with subclinical problems and worries. RESULTS: Results indicate that the majority of the public identifies schizophrenia (88%) and major depression (69%) as mental illnesses and that most report multicausal explanations combining stressful circumstances with biologic and genetic factors. Results also show, however, that smaller proportions associate alcohol (49%) or drug (44%) abuse with mental illness and that symptoms of mental illness remain strongly connected with public fears about potential violence and with a desire for limited social interaction. CONCLUSIONS: While there is reason for optimism in the public's recognition of mental illness and causal attributions, a strong stereotype of dangerousness and desire for social distance persist. These latter conceptions are likely to negatively affect people with mental illness. PMID- 10474549 TI - Who are "the homeless"? Reconsidering the stability and composition of the homeless population. AB - OBJECTIVES: This study assessed the extent to which the use of point-prevalence samples biases conclusions drawn about homeless people. METHODS: Three data sets and published research were used to examine the degree to which conditions leading to point-prevalence bias (turnover in the homeless population, variability in the persistence of homelessness, and associations between personal characteristics and persistence) characterize the homeless population. Results were compared from point-prevalence studies concerning persistence of homelessness and characteristics of homeless people with those from a study of formerly homeless people. RESULTS: Conditions leading to point-prevalence bias strongly characterize the homeless population. Moreover, profiles of homeless people differed dramatically between point-prevalence studies and the study of formerly homeless people. In the former, average duration of homelessness was longer, and samples included higher proportions of men, minorities, non-high school graduates, and people with histories of psychiatric hospitalization, incarceration, and detoxification. CONCLUSIONS: Reliance on point-prevalence samples, when such samples are generalized beyond the currently homeless population, leads to overestimations of the persistence of homelessness, the demographic distinctiveness of the homeless population, and the prevalence of personal disabilities and deviant lifestyles among homeless people. PMID- 10474550 TI - The public's view of the competence, dangerousness, and need for legal coercion of persons with mental health problems. AB - OBJECTIVES: The authors examined Americans' opinions about financial and treatment competence of people with mental health problems, potential for harm to self or others, and the use of legal means to force treatment. METHODS: The 1996 General Social Survey provided interview data with a nationally representative sample (n = 1444). Respondents were given a vignette based on diagnostic criteria for schizophrenia, major depression, alcohol dependence, or drug dependence, or a "control" case. RESULTS: The specific nature of the problem was the most important factor shaping public reaction. Respondents viewed those with "troubles," alcohol dependence, or depression as able to make treatment decisions. Most reported that persons with alcohol or drug problems or schizophrenia cannot manage money and are likely to be violent toward others. Respondents indicated a willingness to coerce individuals into treatment. Respondent and other case characteristics rarely affected opinions. CONCLUSIONS: Americans report greater concern with individuals who have drug or alcohol problems than with persons who have other mental health problems. Evaluations of dangerousness and coercion indicate a continuing need for public education. PMID- 10474551 TI - Exploring the effect of depression on physical disability: longitudinal evidence from the established populations for epidemiologic studies of the elderly. AB - OBJECTIVES: This study examined the effect of depression on the incidence of physical disability and the role of confounding and explanatory variables in this relationship. METHODS: A cohort of 6247 subjects 65 years and older who were initially free of disability was followed up for 6 years. Baseline depression was assessed by the Center for Epidemiological Studies Depression Scale. Disability in mobility and disability in activities of daily living were measured annually. RESULTS: Compared with the 5751 nondepressed subjects, the 496 depressed subjects had a relative risk (95% confidence interval) of 1.67 (1.44, 1.95) and 1.73 (1.54, 1.94) for incident disability in activities of daily living and mobility, respectively. Adjustment for sociodemographic characteristics and baseline chronic conditions reduced the risks to 1.39 (1.18, 1.63) and 1.45 (1.29, 1.93), respectively. Less physical activity and fewer social contacts among depressed persons further explained part of their increased disability risk. CONCLUSIONS: Depression in older persons may increase the risk for incident disability. This excess risk is partly explained by depressed persons' decreased physical activity and social interaction. The role of other factors (e.g., biological mechanisms) should be examined. PMID- 10474552 TI - Effects of health care cost-containment programs on patterns of care and readmissions among children and adolescents. AB - OBJECTIVES: This study examined the effects of a utilization management program on patterns of medical care among children and adolescents. METHODS: From 1989 through 1993, the program conducted 8568 reviews of pediatric patients, ranging in age from birth to 18 years. The program used preadmission and concurrent review procedures to review and certify patients' need for care. This study used multivariate analyses to assess changes in the number of days of inpatient care approved by the program and to determine whether limitations imposed on length of stay affected the risk of 60-day readmission. RESULTS: Concurrent review reduced the number of requested days of inpatient care by 3.2 days per patient. Low birthweight infants and adolescent patients with depression or alcohol or drug dependence accounted for a disproportionate share of the reduction. Patients classified as admitted for medical or mental health care and whose stay was restricted by concurrent review were more likely (P < .05) to be readmitted within 60 days after discharge. CONCLUSIONS: By limiting care through its review procedures, the utilization management program decreased inpatient resource consumption but also increased the risk of readmission for some patients. Continued investigation should be conducted of the effects of cost-containment programs on the quality of care given to children and adolescents, especially in the area of mental health. PMID- 10474553 TI - The extent of drug therapy for attention deficit-hyperactivity disorder among children in public schools. AB - OBJECTIVES: The purpose of this study was to determine the extent of medication use for attention deficit-hyperactivity disorder (ADHD) in southeastern Virginia. METHODS: Students enrolled in grades 2 through 5 in school districts in city A (n = 5767 students) and city B (n = 23,967 students) were included. Nurses recorded students who received ADHD medication in school. RESULTS: The proportion of students receiving ADHD medication was similar in both cities (8% and 10%) and was 2 to 3 times as high as the expected rate of ADHD. Receipt of drug therapy was associated with social and educational characteristics. Medication was used by 3 times as many boys as girls and by twice as many Whites as Blacks. Medication use increased with years in school, and by fifth grade 18% to 20% of White boys were receiving ADHD medication. Being young for one's grade was positively associated with medication use (P < .01). The prevalence of ADHD was 12% in district A, 63% in district B. CONCLUSIONS: These findings suggest that criteria for diagnosis of ADHD vary substantially across US populations, with potential overdiagnosis and overtreatment of ADHD in some groups of children. PMID- 10474554 TI - Minimum-age drinking laws and youth suicide, 1970-1990. AB - OBJECTIVES: This study examined the association between the minimum legal drinking age (MLDA) and suicides among youths aged 18 to 20 years. METHODS: The study used pooled cross-sectional time-series data on youth suicide and the MLDA for the 48 contiguous states in the United States from 1970 to 1990. Poisson regression was used to estimate the association between MLDA and youth suicide. RESULTS: A significant association exists between MLDA and youth suicide. Between 1970 and 1990, the suicide rate of 18- to 20-year-old youths living in states with an 18-year MLDA was 8% higher than the suicide rate among 18- to 20-year-old youths in states with a 21-year MLDA (P < .01). CONCLUSIONS: MLDA is associated not only with the motor-vehicle death rate among youths but also with the rate of youth suicide. We estimate that lowering the drinking age from 21 to 18 years in all states could increase the number of suicides in the 18- to 20-year-old population by approximately 125 each year. PMID- 10474555 TI - Estimating the prevalence of disordered gambling behavior in the United States and Canada: a research synthesis. AB - OBJECTIVES: This study developed prevalence estimates of gambling-related disorders in the United States and Canada, identified differences in prevalence among population segments, and identified changes in prevalence over the past 20 years. METHODS: A meta-analytic strategy was employed to synthesize estimates from 119 prevalence studies. This method produced more reliable prevalence rates than were available from any single study. RESULTS: Prevalence estimates among samples of adolescents were significantly higher than estimates among samples of adults for both clinical (level 3) and subclinical (level 2) measures of disordered gambling within both lifetime and past-year time frames (e.g., 3.9% vs 1.6% for lifetime estimates of level 3 gambling). Among adults, prevalence estimates of disordered gambling have increased significantly during the past 20 years. CONCLUSIONS: Membership in youth, treatment, or prison population segments is significantly associated with experiencing gambling-related disorders. Understanding subclinical gamblers provides a meaningful opportunity to lower the public health burden associated with gambling disorders. Further research is necessary to determine whether the prevalence of disordered gambling will continue to increase among the general adult population and how prevalence among adolescents will change as this cohort ages. PMID- 10474556 TI - Prenatal effects of maternal smoking on daughters' smoking: nicotine or testosterone exposure? AB - OBJECTIVES: The purpose of this study was to specify the effect of prenatal fetal exposure to maternal cotinine and testosterone on daughters' smoking in adolescence and adulthood. METHODS: Longitudinal causal models were estimated among 240 White mother-daughter pairs from the Child Health and Development Study. Mothers and daughters were reinterviewed when daughters were aged 15 to 17 years, and daughters were interviewed at 27 to 30 years of age. Blood samples were obtained from both parents during pregnancy and from adult daughters. RESULTS: Testosterone and smoking were positively correlated among mothers during their pregnancy and among adult daughters. Maternal prenatal cotinine had no direct effect on daughters' smoking; self-reported smoking in pregnancy did have a direct effect. Smoking among daughters during adolescence was determined by maternal prenatal testosterone and self-reported maternal smoking during pregnancy and postnatally. Smoking among adult daughters reflected chronic smoking since adolescence and the continuing effect of postnatal maternal smoking. Prenatal maternal testosterone affected adult daughters' testosterone. CONCLUSIONS: Estimates of the impact of prenatal maternal smoking depend on the measure of smoking. Prenatal testosterone exposure is a previously unrecognized risk factor for smoking among female offspring. PMID- 10474557 TI - The health of grandparents raising grandchildren: results of a national study. AB - OBJECTIVES: This study sought to compare the functional and self-rated health of grandparents raising grandchildren with that of noncaregiving grandparents. METHODS: A secondary analysis of data from the 1992 to 1994 National Survey of Families and Households was conducted. Bivariate and logistic analyses compared 173 custodial and 3304 noncustodial grandparents in terms of functional health limitations, self-rated health, and satisfaction with health. RESULTS: Custodial grandparents were significantly more likely to have limitations in 4 of the 5 activities of daily living (ADLs) examined, with more than half reporting some limitation in 1 of the 5 ADLs. A logistic regression analysis indicated that caregiving grandparents had 50% higher odds of having an ADL limitation. Caregivers were significantly more likely to report lower satisfaction with health, and a statistical trend indicated that the caregivers had lower self rated health. CONCLUSIONS: Further research is needed to determine whether the differences observed reflect artifacts or actual differences in functional abilities and other health measures. The need for policies that support rather than penalize grandparents raising grandchildren is stressed. PMID- 10474558 TI - Fruit and vegetable consumption and prevention of cancer: the Black Churches United for Better Health project. AB - OBJECTIVES: This study assessed the effects of the Black Churches United for Better Health project on increasing fruit and vegetable consumption among rural African American church members in North Carolina. METHODS: Ten counties comprising 50 churches were pair matched and randomly assigned to either intervention or delayed intervention (no program until after the follow-up survey) conditions. A multicomponent intervention was conducted over approximately 20 months. A total of 2519 adults (77.3% response rate) completed both the baseline and 2-year follow-up interviews. RESULTS: The 2 study groups consumed similar amounts of fruits and vegetables at baseline. AT the 2-year follow-up, the intervention group consumed 0.85 (SE = 0.12) servings more than the delayed intervention group (P < .0001). The largest increases were observed among people 66 years or older (1 serving), those with education beyond high school (0.92 servings), those widowed or divorced (0.96 servings), and those attending church frequently (1.3 servings). The last improvement occurred among those aged 18 to 37 years and those who were single. CONCLUSIONS: The project was a successful model for achieving dietary change among rural African Americans. PMID- 10474559 TI - Effects of HIV counseling and testing on sexual risk behavior: a meta-analytic review of published research, 1985-1997. AB - OBJECTIVES: This study examined whether HIV counseling and testing leads to reductions in sexual risk behavior. METHODS: The meta-analysis included 27 published studies that provided sexual behavior outcome data, assessed behavior before and after counseling and testing, and provided details sufficient for the calculation of effect sizes. The studies involved 19,597 participants. RESULTS: After counseling and testing, HIV-positive participants and HIV-serodiscordant couples reduced unprotected intercourse and increased condom use more than HIV negative and untested participants. HIV-negative participants did not modify their behavior more than untested participants. Participants' age, volition for testing, and injection drug use treatment status, as well as the sample seroprevalence and length of the follow-up, explained the variance in results. CONCLUSIONS: HIV counseling and testing appears to provide an effective means of secondary prevention for HIV-positive individuals but, as conducted in the reviewed studies, is not an effective primary prevention strategy for uninfected participants. Theory-driven research with attention given to the context of testing is needed to further explicate the determinants of behavior change resulting from HIV counseling and testing, and the effectiveness of specific counseling approaches. PMID- 10474560 TI - Prevalence and correlates of survival sex among runaway and homeless youth. AB - OBJECTIVE: This study examined the prevalence and correlates of survival sex among runaway and homeless youths. METHODS: A nationally representative sample of shelter youths and a multicity sample of street youths were interviewed. RESULTS: Approximately 28% of street youths and 10% of shelter youths reported having participated in survival sex, which was associated with age, days away from home, victimization, criminal behaviors, substance use, suicide attempts, sexually transmitted disease, and pregnancy. CONCLUSIONS: Intensive and ongoing services are needed to provide resources and residential assistance to enable runaway and homeless youths to avoid survival sex, which is associated with many problem behaviors. PMID- 10474561 TI - Trends in perceived cost as a barrier to medical care, 1991-1996. AB - OBJECTIVES: This study examined trends in perceived cost as a barrier to medical care. METHODS: The Behavioral Risk Factor Surveillance System was used to analyze monthly telephone survey data from 45 states. RESULTS: Overall, the percentage of persons perceiving cost as a barrier to medical care increased from 1991 until early 1993 and then declined to baseline values in late 1996. Perceived cost was a greater barrier in 1996 than in 1991 for persons with low incomes and for those who were unemployed and uninsured. For self-employed persons, percentages increased until mid-1993 and then remained constant. CONCLUSIONS: Further efforts are needed to improve access to medical care for socially disadvantaged populations. PMID- 10474562 TI - Blood pressure level and incidence of myocardial infarction among patients treated for hypertension. AB - OBJECTIVES: This study examined the relationship between achieved blood pressure and risk of myocardial infarction among patients treated for hypertension. METHODS: Blood pressure and other cardiovascular risk factors were assessed among 718 myocardial infarction case patients and 2136 matched controls. RESULTS: Blood pressure level was directly related to risk of myocardial infarction. Patients with treated hypertension who had mild elevations in blood pressure accounted for a larger share of the excess myocardial infarction incidence than those who had higher blood pressure readings. CONCLUSIONS: Achieving normotensive levels in treated hypertensive patients with uncontrolled blood pressure might prevent more than 15% of myocardial infarctions in the treated hypertensive population. PMID- 10474563 TI - Prevalence of self-reported nutrition-related health problems in the Lower Mississippi Delta. AB - OBJECTIVES: The purpose of this study was to assess demographic and geographic differences in prevalence of self-reported nutrition-related health problems in Arkansas, Louisiana, and Mississippi. METHODS: The authors analyzed 1991 and 1993 Behavioral Risk Factor Surveillance System data for adults 18 years or older. RESULTS: Less educated African American women and women of other minority groups who were aged 35 to 64 years reported the highest prevalence of health problems. Geographic differences involved prevalence of hypertension, health status, and insurance status. CONCLUSIONS: Specific demographic subgroups and geographic areas with a high risk of health problems are in particular need of targeted interventions. PMID- 10474564 TI - The Abuela Project: safe cheese workshops to reduce the incidence of Salmonella typhimurium from consumption of raw-milk fresh cheese. AB - OBJECTIVES: A multiagency intervention was implemented in Yakima County, Wash, to reduce the incidence of Salmonella serotype Typhimurium infections resulting from eating queso fresco (fresh cheese) made from raw milk, a traditional food in the Hispanic diet. METHODS: A pasteurized-milk queso fresco recipe with taste and texture acceptable to the Hispanic community was developed. Trained Hispanic volunteers conducted safe cheese workshops, which were attended by more than 225 persons. RESULTS: Workshop participants' acceptance of the new recipe was excellent and positive behavior changes were maintained over 6 months. CONCLUSIONS: Educational interventions in Hispanic communities can reduce the incidence of Salmonella Typhimurium associated with eating queso fresco. PMID- 10474565 TI - Lobbying and advocacy for the public's health: what are the limits for nonprofit organizations? AB - Nonprofit organizations play an important role in advocating for the public's health in the United States. This article describes the rules under US law for lobbying by nonprofit organizations. The 2 most common kinds of non-profits working to improve the public's health are "public charities" and "social welfare organizations." Although social welfare organizations may engage in relatively unlimited lobbying, public charities may not engage in "substantial" lobbying. Lobbying is divided into 2 main categories. Direct lobbying refers to communications with law-makers that take a position on specific legislation, and grassroots lobbying includes attempts to persuade members of the general public to take action regarding legislation. Even public charities may engage in some direct lobbying and a smaller amount of grassroots lobbying. Much public health advocacy, however, is not lobbying, since there are several important exceptions to the lobbying rules. These exceptions include "non-partisan analysis, study, or research" and discussions of broad social problems. Lobbying with federal or earmarked foundation funds is generally prohibited. PMID- 10474566 TI - A Safe Haven: an innovative approach to residential treatment of substance abuse. PMID- 10474567 TI - Countering pro-tobacco influences at the racetrack. PMID- 10474568 TI - Awareness and motivation of Japanese donors of blood for research. PMID- 10474569 TI - The effect of plain packaging on response to health warnings. PMID- 10474570 TI - Sex differences in cardiovascular disease mortality. PMID- 10474571 TI - Household gun ownership. PMID- 10474572 TI - Parental attitudes and childhood immunization. PMID- 10474573 TI - Promoting an evidence base for general practice. PMID- 10474574 TI - Acne and acne scarring: why should we treat? PMID- 10474575 TI - Medicine and palliative care. PMID- 10474576 TI - Oral agents in type 2 diabetes: where to now? PMID- 10474577 TI - Asthma in general practice: action plans or planned actions. PMID- 10474578 TI - Randomised controlled trial of general practice based asthma clinics. AB - OBJECTIVE: To compare the effects on asthma morbidity of asthma clinics based in general practice with standard general practice care. DESIGN AND SETTING: A randomised controlled trial in eight general practices. Patients, general practitioners and outcomes assessors were not blinded to treatment allocation. PARTICIPANTS: 195 patients with asthma aged 5-64 years; 191 completed the trial. INTERVENTION: Three asthma clinic sessions over six months involving nurse counselling, education about asthma management, spirometry and consultation with the general practitioner. MAIN OUTCOME MEASURES: Patients reporting days lost from work or school, number of days lost, the presence of morning or nocturnal asthma symptoms, use of an action plan, medication use, current smoking, hospitalisation, and emergency visits. RESULTS: Asthma clinics were associated with a greater reduction in nocturnal symptoms, an increase in the ownership of peak flow meters and an increase in the number of patients commencing or resuming smoking. Both control and intervention groups showed similar improvement in days lost from work or school, the presence of symptoms, use of an action plan and taking reliever medication. CONCLUSION: Our study does not show that asthma clinics are more effective than standard general practice care in reducing asthma morbidity. It is uncertain how much of the improvement in outcomes was due to the asthma clinic, the influence of the study itself upon patients and practitioners, or other factors, such as the tendency for a patient's asthma management to improve over time. PMID- 10474579 TI - Comparison of patients with asthma managed in general practice and in a hospital clinic. AB - OBJECTIVES: To compare knowledge and attitudes about asthma, self-management skills and impact of asthma on quality of life between patients managed in general practice (GP) and in a hospital clinic. DESIGN: Cross-sectional survey with six months' follow-up. PATIENTS AND SETTING: 105 adults with asthma: 61 from the Alfred Hospital Asthma and Allergy Clinic, Melbourne, and 44 from nearby general practices, in 1994-1995. MAIN OUTCOME MEASURES: Patient sociodemographic and clinical characteristics; patient knowledge, attitudes and beliefs about asthma; self-management skills; and impact of asthma on quality of life. RESULTS: GP patients were more educated (P = 0.04) and more likely to smoke (P = 0.04) and to have mild asthma (P = 0.04) than hospital patients; they were less likely to use theophylline (P = 0.006) and to have exercise limitation (P = 0.03), and had fewer previous hospital admissions (P = 0.01). Impact of asthma on quality of life was greater in the hospital group than in the GP group. At baseline, the GP group were less likely to have written asthma action plans (P = 0.018), and were less able to manage rapid onset attacks than the hospital group (P = 0.02). More subjects in the hospital group than the GP group felt their asthma was severe (P = 0.02) and were optimistic about their asthma improving (P = 0.03). GP patients increased their knowledge about asthma significantly (P = 0.002) over six months. CONCLUSIONS: Patients with asthma managed in general practice and in hospital differ in clinical parameters, quality of life and attitudes to asthma. Future educational initiatives should take such differences into account. PMID- 10474580 TI - General practice consultations: quality time? PMID- 10474581 TI - Consultation length and chronic illness care in general practice: a qualitative study. AB - OBJECTIVE: To study the relationship between duration and content of general practice consultations for patients with chronic conditions. DESIGN: A qualitative analysis of transcripts of consultations. The major themes and concepts of psychosocial support were identified and coded using the Ethnograph computer package. SETTING: A mix of rural and urban general practices in two States of Australia in 1993-1994. PARTICIPANTS: 14 selected general practitioners and 50 of their patients with complex chronic conditions. RESULTS: Transcriptions of 106 consultations were analysed. General practitioners (GPs) led most consultation dialogue and emphasised disease management. The major themes were provision of information by the GP, review of treatment by the GP, review of illness by the GP, and description and explanations of their illness by patients (patient narrative). The first three themes predominated in consultations of all lengths. Longer consultations (20 minutes and over) contained more dialogue initiated by patients and more patient narrative about living with their illness. CONCLUSIONS: Patients with complex chronic conditions may require longer consultations to allow adequate time for review of their illness and treatment as well as an opportunity to raise issues and concerns about their illness, its impact on their lives and their personal management strategies. Longer consultations may thus provide the mechanism for what has been described as patient "enablement". PMID- 10474582 TI - Vaccines and the cold chain: is it too hot ... or too cold? PMID- 10474583 TI - Electronic temperature monitoring and feedback to correct adverse vaccine storage in general practice. AB - OBJECTIVE: To evaluate the effect of direct feedback of temperature conditions followed by a telephone educational questionnaire to correct adverse vaccine storage, and to determine the consistency of vaccine storage conditions at provider sites over six months. PARTICIPANTS: 32 general practitioner vaccine providers in metropolitan Adelaide. DESIGN: Temperatures were monitored for 14 days using electronic temperature monitors, with repeat monitoring at two and six months. Feedback was given to vaccine providers and an educational questionnaire was administered after initial monitoring. MAIN OUTCOME MEASURE: Number of sites with more than 20% of recordings < 2 degrees C or > 8 degrees C and/or more than 5% of recordings < 0 degree C or > 15 degrees C. RESULTS: 13 (41%) sites had initially suboptimal storage. Following the interventions, storage was corrected in all but three (23%) of these sites. Only 10 (52%) of the 19 initially optimal sites had consistent optimal storage at two and six months. CONCLUSION: Electronic monitoring with direct feedback of storage temperatures was effective in correcting adverse storage at most suboptimal sites. Vaccine storage at initially optimal sites was not consistent. Our findings have important implications for further research and public health measures aimed at correcting and maintaining optimal vaccine storage. PMID- 10474584 TI - Shared care through divisions of general practice: moving forward. AB - Divisions of general practice have established a wide variety of structured programs to support shared care over the past six years. Differences in the adoption and continuation of these programs appear to be related to a number of factors, including local opportunities and external funding and demands on divisions, relationships and the capacity of the divisions and their health service partners. PMID- 10474585 TI - Out-of-surgery care: will there be a role for Australian general practitioners? PMID- 10474586 TI - Clinical practice guidelines and general practice: the sleeping giant in Australian healthcare integration? PMID- 10474587 TI - Medical care in aged-care facilities: new directions. PMID- 10474588 TI - Necrotising arachnidism. PMID- 10474590 TI - As doctors, we are healers. PMID- 10474589 TI - Acute and recurrent skin ulceration after spider bite. AB - We reviewed the records of the Australian Venom Research Unit and The Alfred Hospital Department of Hyperbaric Medicine from January 1992 to July 1998 and found 15 cases of skin ulceration after spider bite that could be followed up with the patient and the treating physician. Fourteen patients had skin ulceration attributed to white-tailed spider bites but in only three was this confirmed. One patient had skin necrosis after a confirmed black house spider bite. Recurrent skin ulceration occurred in nine of the 15 patients. PMID- 10474591 TI - Problems with new criteria for diagnosis of diabetes mellitus. PMID- 10474592 TI - Problems with new criteria for diagnosis of diabetes mellitus. PMID- 10474593 TI - Problems with new criteria for diagnosis of diabetes mellitus. PMID- 10474594 TI - Severe allergen-induced asthma despite the use of Buteyko breathing technique. PMID- 10474595 TI - Hospital in the home: a randomised controlled trial. PMID- 10474596 TI - Primary care psychiatry is not specialist psychiatry in general practice. PMID- 10474597 TI - Abnormal involuntary movements in people with schizophrenia in the community. PMID- 10474598 TI - Should smoking be an indication for lipid-lowering therapy? PMID- 10474599 TI - Vancomycin-resistant enterococci: causes and control. PMID- 10474600 TI - Who should be screened for thyroid dysfunction? PMID- 10474601 TI - Management of cerebral aneurysms: current best practice. PMID- 10474602 TI - HIV vaccines--promise and directions. PMID- 10474603 TI - Vancomycin and teicoplanin use in Victorian hospitals. The Victorian Drug Usage Evaluation Group. AB - OBJECTIVE: To determine patterns of prescribing of glycopeptide antibiotics (vancomycin and teicoplanin) in Victorian hospitals and identify areas for targeted intervention. DESIGN: A concurrent, observational, multisite evaluation of drug use. SETTING: Thirty-five Victorian hospitals, 1-14 September 1997. STUDY POPULATION: Patients commencing a glycopeptide antibiotic course. MAIN OUTCOME MEASURES: Rate of glycopeptide antibiotic use; indications; duration of use; main hospitals using glycopeptide antibiotics. RESULTS: 293 patients (269 adults and 24 neonates) commenced on 302 glycopeptide antibiotic courses: 296 intravenous (i.v.) vancomycin courses and three each of oral vancomycin and parenteral teicoplanin. The overall rate of use was 10.3 courses per 1000 inpatient separations. Of 271 i.v. vancomycin courses for adults, 176 (65%) were for treatment--120 empirically. The median duration of treatment courses was 4.7 days (interquartile range, 2.0-8.2 days). A flucloxacillin-resistant organism was confirmed for 44% of treatment courses. Ninety-five i.v. vancomycin courses were for prophylaxis, including for cardiac (54%) and vascular surgery (21%); 82% of prophylactic courses were administered for less than 24 hours. Of all the glycopeptide antibiotic courses, 69% were administered at five major metropolitan hospitals. CONCLUSIONS: Glycopeptide antibiotic use in Victoria is concentrated in the major metropolitan hospitals. Prolonged durations of vancomycin therapy, including for surgical prophylaxis and empirical therapy not subsequently confirmed by microbiology findings, would be suitable targets for interventional strategies. PMID- 10474604 TI - Outcome of a screening program for vancomycin-resistant enterococci in a hospital in Victoria. AB - OBJECTIVE: To screen for faecal colonisation with vancomycin-resistant enterococci (VRE) among potentially at-risk patients. DESIGN: Infection control screening program. SETTING: Monash Medical Centre (a tertiary care hospital), Melbourne, Victoria, in the seven months from June 1997. PATIENTS: Patients in the Renal, Oncology and Intensive Care (ICU) Units. MAIN OUTCOME MEASURES: Presence of VRE in a rectal swab or faecal specimen taken at admission and at regular intervals during inpatient stay; presence of vancomycin-resistance genes (vanA, vanB and vanC) assessed by polymerase chain reaction (PCR); genetic clonality of isolates assessed by pulsed-field gel electrophoresis (PFGE). RESULTS: 574 patients (356 renal, 134 ICU and 84 oncology) were screened; 12 were colonised with VRE--nine renal inpatients, two having peritoneal dialysis or incentre haemodialysis, and one ICU patient. Nine isolates were Enterococcus faecalis (seven positive for vanB and two negative for all three resistance genes) and three were Enterococcus faecium (all positive for vanB). Eight were high-level gentamicin resistant. PFGE suggested genetic clonality between the index isolate and five other isolates from renal patients. No specific clinical practice was associated with VRE colonisation. Attempts to clear rectal carriage with oral ampicillin/amoxycillin or bacitracin were of limited success. Although antibiotic prescribing in the Renal Unit was generally consistent with defined protocols, use of vancomycin and third-generation cephalosporins has been further restricted. CONCLUSIONS: Renal inpatients in our institution appear most at risk of VRE colonisation (4.6% overall) and therefore of VRE infection. Routine screening, especially of potentially high-risk patients, should be considered in major Australian hospitals. PMID- 10474605 TI - Australian suicide trends 1964-1997: youth and beyond? AB - OBJECTIVE: To examine Australian suicide rates across all ages, and compare Australian rates with those of other Western nations. DESIGN: Australian Bureau of Statistics data were used to examine Australian suicide trends, 1964-1997, by age and sex. For comparison, suicide rates of 22 other Western nations, 1990 1994, were obtained from the World Health Organization. RESULTS: Australian suicide rates for males 15-24 years and 25-34 years rose from 1964-1997. Comparable rates for females showed no significant change. Suicide rates for several of the older age and sex groups declined over this period. Comparison with suicide rates of other Western nations showed that, while Australian youth suicide rates are relatively high, this is not the case for older age groups. Australian suicide rates are higher than those in the European nations of origin of our major migrant groups, but similar to those of other Western nations also recently colonised by Europeans (Canada, the United States and New Zealand). CONCLUSIONS: Priorities for suicide prevention in Australia are correctly concentrated on youth, but the targeted age range should be extended to include men aged 25-34 years. A comprehensive policy should also not neglect the needs of other age groups. Further epidemiological study of national and international data may suggest new approaches to suicide prevention. PMID- 10474606 TI - Diaphragmatic endometriosis. AB - Endometriosis is a relatively common condition usually found in the pelvis. However, lesions do occur outside the pelvis and, more rarely, in the upper abdomen. In the case reported here, the patient presented with chronic right shoulder tip pain. The diagnosis of extrapelvic endometriosis is often not considered in such circumstances. This patient's symptoms were relieved by surgical excision of the diaphragmatic lesion. PMID- 10474608 TI - Medicolegal activity in obstetrical and gynaecological ultrasound. AB - We surveyed obstetricians who are members of the Australian Association of Obstetrical and Gynaecological Ultrasonologists for details of actions in which they had been asked to give an expert opinion between 1993 and 1998. We uncovered 50 cases, most of which involved missed fetal anomalies. Our findings suggest there is considerable medicolegal activity in obstetrical and gynaecological ultrasound in Australia. PMID- 10474607 TI - Vancomycin-resistant enterococci and use of avoparcin in animal feed: is there a link? AB - Australia is unique among Western countries in allowing animal use of this vancomycin-like antibiotic. PMID- 10474610 TI - Universal recommendations for sodium intake should be avoided. PMID- 10474609 TI - The acute coronary syndromes: myocardial infarction and unstable angina. PMID- 10474612 TI - Heroin, HIV and harm in New York and Sydney. PMID- 10474613 TI - The force of numbers: why hepatitis C is spreading among Australian injecting drug users while HIV is not. PMID- 10474614 TI - A complication of sclerotherapy. PMID- 10474615 TI - Rethinking the early childcare agenda. PMID- 10474616 TI - Should research ethics change at the border? PMID- 10474617 TI - Changing attitudes towards antiretroviral treatment of HIV infection: a prospective study in a sample of Dutch general practitioners. AB - This study investigated the attitude towards antiretroviral therapy (ART) for HIV infection over time, in a sample of Dutch general practitioners (GPs). Twenty-one GPs, of which 16 were practising in Amsterdam, completed multiple questionnaires on HIV-related topics between April 1995 and March 1997. In 1995, only 10% had a positive attitude towards treatment of asymptomatic persons with a CD4+ > 300 x 10(6)/l; at the end of the study 43% had. In 1995, 57% had a positive attitude towards treatment of asymptomatic persons with a CD4+ < or = 300 x 10(6)/l, and 52% towards treatment of symptomatic patients with a CD4+ < or = 400 x 10(6)/l; heterosexual GPs more often had a positive attitude as compared to homosexual GPs (p = 0.005 and p = 0.01, respectively). At the end of the study the proportions of GPs with a positive attitude had increased from 57 to 81% and 52 to 95%, respectively. The risk of adverse effects, strict dose regimens and medicalization were regarded as the main disadvantages of the current treatment strategy. The conclusion is that the attitude towards ART has become more positive since 1995. At the beginning of 1997, however, there still existed reservations about treatment of asymptomatic persons with CD4+ cell counts > 300 x 10(6)/l. PMID- 10474618 TI - Use of protease inhibitors among persons with AIDS in Los Angeles County. AB - We evaluated the access to, and the factors associated with, protease inhibitor use among persons with AIDS in Los Angeles County. A population-based sample of adult persons with AIDS is routinely interviewed in Los Angeles County and includes a 30% random sample of men who have sex with men and all other persons reported with AIDS. Since May of 1996, all participants were asked if their physician had ever prescribed a protease inhibitor for their use. The possible association between protease inhibitor use and sociodemographic, temporal and health care factors was assessed for the 12-month period May 1996 through April 1997. Logistic regression was employed for multivariate analysis. Over the 12 month study period, 61.7% (209/339) persons interviewed reported that their physician had prescribed a protease inhibitor as part of their therapy. In bivariate analysis, treatment with protease inhibitor use was more common for whites (71.4%) and US-born Latinos (68.2%) than blacks (53.4%) and foreign-born Latinos (56.6%), among person of higher income (71.2%) than lower (< $10,000) income (50.3%), in those who reported having insurance (66.7%) than those uninsured (47%) and among persons receiving care at private clinics (86.4%) than at HMOs (63.4%) or public clinics (55.2%). An increasing trend of protease inhibitor use with higher educational level and declining CD4+ count was observed. A temporal increase was noted and this trend was most pronounced for persons receiving care at public clinics. In multivariate analysis, persons receiving care at private facilities (adjusted OR = 2.9, 95% CI 1.0, 8.2) and those with higher incomes (adjusted OR = 2.5, 95% CI 1.5, 4.3), were more likely to report that their physician had prescribed a protease inhibitor. The effect of facility type was modified by time. During the first six months of the study period (May 1996-October 1996) persons with AIDS receiving care at public facilities and HMO sites were substantially less likely to report having been offered a protease inhibitor (adjusted OR = 0.13, 95% CI 0.03, 0.58 and adjusted OR = 0.23, 95% CI 0.05, 1.2, respectively). However, no significant facility specific differences were observed over the last six-month period (November 1997 April 1997) evaluated. Our findings suggest that substantial differences exist in the prescribing and use of protease inhibitors among persons with AIDS in Los Angeles County. Several factors, including facility of HIV care, calendar time, income, education level and level of immunosuppression were independently associated with protease inhibitor use and suggest the existence of important barriers to access. Efforts should be made to identify and remove barriers that will ensure the widest possible access to protease inhibitors for patients with a clinical indication for their use. PMID- 10474619 TI - AIDS diagnosis and depression in the Multicenter AIDS Cohort Study: the ameliorating impact of pet ownership. AB - The impact of pet ownership on depression was tested among a sample of gay and bisexual men (n = 1,872). Multivariate analyses, controlling for demographics and baseline depressive symptomatology, showed that neither pet ownership nor the presence of HIV infection was associated with depression. Depression was influenced by the presence of AIDS and by having relatively few confidants. Analyses among HIV-infected men only showed that persons with AIDS who owned pets reported less depression than persons with AIDS who did not own pets. This beneficial effect of pet ownership occurred principally among persons who reported fewer confidants. These results suggest that by enhancing companionship for some HIV-infected persons, pets may buffer the stressful impact of AIDS. PMID- 10474620 TI - Measuring the stress associated with caring for clients with HIV/AIDS. AB - The AIDS-stress scale (Pleck et al., 1988) provides measures of the sources of stress faced by health care workers caring for people with AIDS. The aim of the study was to consider the utility of the scale as a means of identifying groups of health care workers who would benefit from intervention. Data were collected from a sample of Scottish health care workers (n = 140), all with known contact with clients with HIV or AIDS within the year prior to data collection. Three factors were derived from the AIDS-stress scale: 'lack of knowledge', 'discomfort' and 'work load'. Each showed a different pattern of association with occupational, training and attitudinal measures. The factor scores had some degree of concurrent validity and were not, in the main, associated with the desire to give socially desirable responses. The results were discussed in relation to appropriate interventions. PMID- 10474621 TI - Heterosexual couples confronting the challenges of HIV infection. AB - Couples confronted with HIV infection face significant challenges. Little is known about the impact of HIV on heterosexual couples who account for the vast majority of cases worldwide and an increasing proportion of cases in the USA, especially among women. In this study, analysis of data collected on HIV discordant couples participating in a ten-week support group revealed four major groups of issues: (1) dealing with the emotional and sexual impact on the relationship; (2) confronting reproductive decisions; (3) planning for the future of children and the surviving partner; and (4) disclosure of the HIV infection to friends and family. These findings have implications for the design of interventions to enhance adaptation to HIV for discordant couples. PMID- 10474622 TI - Women who are HIV infected: the role of religious activity in psychosocial adjustment. AB - The role of religious activity in the psychosocial adjustment of 205 inner-city African-American women, one-half of whom are HIV infected, was examined. Those who were HIV infected reported praying more but viewed prayer as less effective in coping with a chronic illness. Frequency of prayer predicted optimism about the future, whereas religious activity was not related to current depressive symptoms. PMID- 10474623 TI - 'In an important way, I did die': uncertainty and revival in persons living with HIV or AIDS. AB - This study reports the revival experiences of persons who once were reconciled to their death from HIV/AIDS but who, as a result of dramatic treatment responses, now believe they may survive (popularly known as the Lazarus Syndrome). A purposive sample of men and women living with HIV infection or AIDS were interviewed in six focus groups. As part of a larger study of uncertainty in HIV illness, participants described their uncertainty accompanying renewed health and a return to the joys and problems of continued life. While new discoveries about the disease and exciting antiretroviral therapies hold the promise of improved survival, ambiguity about the durability of treatment response and ultimate survival contribute to the level of uncertainty with which a patient must cope. The experience of uncertainty in the narratives about revival involved renegotiation. Participants described physical renewal as an unexpected new stressor forcing them to renegotiate: (a) feelings of hope and future orientation, (b) social roles and identities, (c) interpersonal relations, and (d) the quality of their lives. Implications for prevention, practice, research and theory are presented and suggestions for education and assistance are offered. PMID- 10474624 TI - Parallel accounts? Discrepancies between self-report (diary) and recall (questionnaire) measures of the same sexual behaviour. AB - Questionnaires and diaries have complementary biases and advantages for obtaining information on sexual behaviour but self-completed sexual diaries have the advantage of reducing retrospective bias. In a validation study of homosexual behaviour, sexual diary counts and subsequent questionnaire estimates (together with ratings of the certainty of the estimates) referring to the same month are compared and the discrepancies analyzed. Main findings include: questionnaire data yield consistently higher average estimates than diary counts, but have the same ordinal profile; individual difference (diary-questionnaire) scores show that 55% of questionnaire estimates of acts are higher than diary counts, 20% are identical and 25% are under-estimates; discrepancies are differentially located in different sexual acts. Masturbation and fellatio are systematically over estimated in questionnaires and anal intercourse without a condom is the major source of inaccuracies, but in different directions: active partners under estimate and passive partners over-estimate the amount of highest-risk sex. A strategy of joint use is discussed. PMID- 10474625 TI - Nature and consequences of stressful life events in homosexual HIV-positive men: a review. AB - A great deal of research has been conducted into the stressful experiences that befall HIV-infected individuals. In this review we have focused on the work that has been conducted with HIV-positive homosexual males and have examined issues concerning the measurement, nature and consequences of stressful life events. It is apparent that, in consort with most other life events research, the measurement of events in this patient group is affected by methodological difficulties. Such difficulties may have, in particular, led to the observation by some investigators that HIV-positive homosexual men experience greater numbers of such events than their HIV-negative counterparts. It is also evident that, while some of these events are typical of those faced by individuals with other life threatening conditions, others are peculiar to this patient group. Research into the consequences of these events for emotional and behavioural functioning indicate that the effects are far-reaching. Similarly, data on the consequences of stressful events for immune function suggest that they may play a role in the progression of the disease, although these data are equivocal. PMID- 10474626 TI - UNAIDS/WHO global AIDS statistics. PMID- 10474627 TI - HIV risk behaviours of current sex workers attending syringe exchange: the experiences of women in five US cities. AB - Existing research indicates that sex workers who inject drugs are vulnerable to HIV infection through both risky sexual and drug use practices. This study is the first attempt to learn whether this increased risk persists among current sex workers who participate in syringe exchange programmes (SEPs). With data from interviews with randomly selected participants in five US cities, we compared the demographic characteristics, sexual risk behaviours, drug use practices, emotional and physical health, and SEP utilization patterns of currently active female sex workers who attend SEPs with female SEP participants who do not engage in sex work. Data indicate that women enrolled in SEPs who were currently trading sex typically reported greater HIV risk than women non-sex workers. Current sex workers reported higher levels of risk for every drug risk variable examined in bivariate analysis. They were more likely than other women to inject with a syringe previously used by someone else, to inject daily and to attend shooting galleries; they were less likely to use a condom with their primary partners and to report higher levels of psychological distress than their counterparts. The relationship between sex work status and risky injection practices persisted when potential confounders were controlled for in multivariate analysis. SEPs can serve a pivotal role in providing sex workers with services and referrals which would help them reduce risk behaviours. PMID- 10474628 TI - Factors associated with gay men's sexual behaviours and risk on holiday. AB - Findings are presented from a questionnaire survey of 562 gay men resident in southern England on their sexual behaviour in the context of holidays. Data were gathered during summer 1996 by means of a self-completion questionnaire; 395 men reported at least one holiday undertaken in 1996 and the analysis focuses on the first or only holiday described. Of 391 men giving details of sexual activity, 187 reported sex with a new partner, 113 men reported penetrative sex and 17 reported penetration without consistent use of condoms. Sexual activity on holiday was predicted by: being on holiday alone or with friends, taking condoms, being motivated by 'gay social life and sex' in planning a holiday, higher expectations of sexual activity/risk and a higher number of new partners at home. Penetrative sex was predicted by: more sexual partners on holiday, taking condoms and higher expectations of sexual activity/risk. Unprotected sex was predicted by: not taking condoms, higher expectations of sexual activity/risk and positive HIV status. Survey findings are discussed in relation to previous research, and implications for travel-related HIV prevention initiatives targeting gay men are explored. PMID- 10474629 TI - HIV risk behaviours among male-to-female transgenders in comparison with homosexual or bisexual males and heterosexual females. AB - Issues and concerns among six directors, one counsellor and one consultant for transgender programmes in San Francisco are described. Additionally, 173 clients who were enrolled in programmes in eight AIDS service organizations were interviewed using a structured survey questionnaire. This study examined the relationships between HIV risk behaviours and social and cognitive factors among transgendered females (male-to-female transgenders) (n = 25) in comparison with homosexual or bisexual males (n = 122) and heterosexual females (n = 26). Transgendered females engaged in riskier behaviours than the other groups in terms of the number of sex partners in the past 30 days and the past six months, commercial sex activities, and having a steady sex partner who injected drugs. Adverse socioeconomic conditions and transgender-specific risk behaviours such as injecting hormones in relation to HIV risk behaviours must be targeted by future intervention studies. PMID- 10474630 TI - Social support and unsupportive social interactions: their association with depression among people living with HIV. AB - Although numerous studies of people living with HIV have focused on positive social support, researchers have directed surprisingly little attention to the nature and effects of negative social interactions in this population. Based on data from a diverse sample of people with HIV (N = 271), we conducted a factor analysis to develop the HIV version of the Unsupportive Social Interactions Inventory (USII). Four types of unsupportive or upsetting responses that an HIV positive person might receive from others were identified: insensitivity, disconnecting, forced optimism and blaming. In analysis with a sub-sample of 96 people with HIV, unsupportive social interactions were only moderately correlated with social support, suggesting that these constructs are relatively independent. Using hierarchical regression analysis, we found that unsupportive social interactions predicted a significant amount of the variance in depression, beyond the variance accounted for by physical functioning and positive social support. Partial correlations indicated that the relationship between unsupportive social interactions and depression was not an artifact of trait negative affectivity. Favourable evidence of the USII's reliability and validity suggests that the instrument provides a useful tool for assessing unsupportive social interactions experienced by people with HIV. Practical implications of the findings and directions for future research are discussed. PMID- 10474631 TI - Patient satisfaction with HIV service provision in NPMS hospitals: the development of a standard satisfaction questionnaire. NPMS Steering Group. AB - A self-completion satisfaction questionnaire evaluating the standard of care of HIV outpatient services was developed as part of the National Prospective Monitoring System on the Use, Cost and Outcome of HIV Service Provision in English Hospitals (NPMS). The questionnaire was designed in conjunction with service users and health care professionals, and piloted in three London and three non-London HIV clinics. In addition to testing alternative methods of administering the questionnaire, the pilot provided satisfaction scores on a variety of aspects of service provision for participating clinics. The questionnaire was completed by 548 respondents and was most effectively collected using a sealed box in the clinic waiting area. Mean satisfaction scores for the attitude and skills of staff members was 4.7 (95% CI 4.6-4.7) but satisfaction scores were significantly lower for the clinic environment with a mean of 4.1 (95% CI 4.1-4.2). Satisfaction scores did not differ significantly by gender, age, sexual orientation, ethnic group, employment status or severity of symptoms. London respondents were more satisfied with the clinic environment and seeing preferred members of staff than their non-London counterparts, however there were no other differences between clinics. The questionnaire functioned well in practice and provided meaningful and useful information for individual clinics as well as at aggregate level. PMID- 10474632 TI - Use of chart reminders for physicians to promote discussion of advance directives in patients with AIDS. AB - To determine if use of a physician chart reminder improves the rate of physician initiated discussion and subsequent completion of advance directives (ADs) in patients with AIDS, a controlled study was conducted with 74 patients with AIDS and ten physicians providing primary care at a university-based hospital clinic. Chart reminders were placed on medical records of intervention patients at each primary care clinic visit for six months. Twelve out of 39 (31%) reminder group patients, but only three out of 35 (9%, p = 0.02) control patients underwent AD discussion with physicians. Further, more subjects in the reminder group completed ADs (28% versus 9%, p = 0.03). Controlling for demographic and clinical factors, only assignment to reminder group was associated with discussion and completion of ADs. Physician-level analysis showed that the effect was physician dependent. Physician chart reminders are an effective tool for promoting discussion and completion of ADs in patients with AIDS although the effect is physician-dependent. PMID- 10474633 TI - Reluctance amongst nurses and doctors to care for and treat patients with HIV/AIDS. AB - A survey was carried out, using a questionnaire and semi-structured interviews, to examine why nurses and doctors might be reluctant to care for patients with HIV/AIDS. The findings provided four main themes: perceptions about sexual promiscuity; blaming certain patients with HIV/AIDS; belief in the right to refuse to provide care; and discriminatory care. The paper concludes by acknowledging the limitations of the study and examining the implications of the findings for clinical practice. PMID- 10474634 TI - Predictors of outpatient medical appointment attendance among persons with HIV. AB - Non-adherence to medical regimens is a critical threat to the health of HIV infected individuals. Patients who do not adhere to routine medical care cannot fully benefit from the increasingly efficacious treatments available to them. Consistent attendance at medical appointments plays a central role in both prolonging life and enhancing quality of life for persons living with HIV/AIDS. By identifying why many patients do not reliably attend medical services, interventions can be undertaken to improve appointment-keeping. The primary purpose of the present study was to identify factors predictive of HIV-related medical appointment attendance. One-hundred-and-forty-four outpatients in a public hospital ambulatory care HIV clinic were followed for seven months. Demographic, medical care and psychosocial factors were measured in order to prospectively predict the percentage of missed clinic appointments by persons with HIV disease. Greater outpatient appointment non-attendance was associated with younger age, minority status, less severe illness and lower perceived social support. Treatment duration, provider consistency, hopelessness and religious coping did not emerge as significant predictors of outpatient appointment-keeping in this sample. Practical and theoretical implications of these findings are discussed in light of recent medical advances in HIV/AIDS care. PMID- 10474635 TI - Young people and HIV in Cambodia: meanings, contexts and sexual cultures. AB - Like many other countries in South East Asia, Cambodia is experiencing a rapidly developing AIDS epidemic. Groups reported as being particularly seriously affected include sex workers and their clients. Young people too may be at heightened risk: some young women find sex work a lucrative option in the context of low wages and poor employment opportunities, and some young men pay for sex either as individuals or as part of group socializing. These same young men may subsequently have sex with other partners, thus extending networks of transmission. While there is limited knowledge about the form of such sexual networks, little is known about the meanings that underpin young people's sexual relations and partnerships, the sexual identities associated with such meanings, and prevailing socio-sexual cultures. This paper reports on findings from an in depth qualitative study conducted among two groups of young people: one urban, the other rural. Following an initial Rapid Assessment Process, data was collected via individual interviews, focus group interviews and participant observation. The research team included young people themselves. Data is presented on dominant discourses about sex and sexuality in Cambodia; contemporary patterns of sexual behaviour; sexual meanings and sexual practices; sexual relations among young people involving payment; and sexual relations not involving payment. The implications for more effective HIV prevention efforts are discussed. PMID- 10474636 TI - Anaemia in acute, Plasmodium falciparum malaria in children from Orissa state, India. AB - The severity of anaemia associated with acute, Plasmodium falciparum malaria and the extent to which haemolysis, bone-marrow suppression, and pre-existent iron deficiency contribute to the anaemia were assessed in 102 Indian children aged 2 12 years. Blood haemoglobin (Hb), plasma unconjugated bilirubin and haptoglobin, serum iron and transferrin concentrations and transferrin saturation, red cell morphology and reticulocyte response were investigated in the patients and in 50 control children. Twenty-three patients with severe anaemia (< 70 g Hb/litre) were investigated further, by bone-marrow biopsy followed by iron staining of sections or touch smears of the biopsy material. There was evidence of haemolysis in the malaria cases: in the peripheral blood smears and the significantly higher plasma concentrations of unconjugated bilirubin, lower plasma concentrations of haptoglobin and lower blood concentrations of Hb than those seen in the controls. Haemoglobin concentration correlated directly with haptoglobin (r = 0.489; P < 0.001) and inversely with unconjugated bilirubin in malaria patients (r = -0.526; P < 0.001) but not in controls (r = -0.140 and -0.061, respectively). Parasitaemia (parasites/microliter) was not significantly correlated with Hb, haptoglobin or unconjugated bilirubin. Compared with the earlier samples, follow up samples from the patients, collected 2 weeks after discharge from hospital and antimalarial therapy, showed significant increase in Hb, haematocrit, haptoglobin and decreases in both total and unconjugated bilirubin. There was evidence of hypercellularity and mild-moderate erythroid hyperplasia, mainly of normoblastic maturation with adequate reticulocyte response, in the bone-marrow samples from the cases of severe anaemia; dyserythropoiesis was only noticed in one case and no stainable iron was detectable in 17 of the 23 cases. These observations indicate that haemolysis is the prime cause of the anaemia seen in acute falciparum malaria, although destruction of parasitised erythrocytes is not the sole cause of the haemolytic process. Bone-marrow suppression appears to have an insignificant role but pre-existent iron deficiency aggravates the severity of the anaemia. PMID- 10474637 TI - An analysis of the determinants of anaemia in pregnant women in rural Malawi--a basis for action. AB - Haematological data are presented on 4104 pregnant women attending the antenatal care facilities of two hospitals in a rural area in southern Malawi. In this area, malaria transmission is perennial and there is a high prevalence of HIV infection. The local women are exposed to drought and food shortages but experience high fertility rates. Mean (S.D.) haemoglobin (Hb) concentration was significantly lower in the primigravidae [8.7 (1.60 g/dl] than in the secundigravidae [9.1 (1.5) g/dl; P < 0.0001] or multigravidae [9.2 (1.5) g/dl; P < 0.0001]. Primigravidae also experienced significantly more iron deficiency and malaria than secundi- or multi-gravidae. For all parity groups, the lowest mean Hb levels were observed between 26-30 weeks' gestation. In primigravidae peak prevalence of malaria occurred between 16-20 weeks' gestation (38.6%) and peak prevalence of moderately severe anaemia (< 8 g Hb/dl) between 26-30 weeks' (35.7%). Multigravidae showed little variation in the prevalence of anaemia, iron deficiency and malaria with gestational age. Peak prevalences of malaria were observed in April, in the post-rainy season, with values of 51.4%, 56.0% and 25.3% for primi-, secundi- and multi-gravidae, respectively. Peak prevalences of iron deficiency occurred between April and May and those of moderately severe anaemia between May and June. Mean Hb was lower in adolescent primigravidae than in any other group of pregnant women [8.6 (1.5) g/dl], including the non adolescent primigravidae [8.9 (1.6) g/dl; P = 0.008]. Other factors significantly associated with increased risk of moderately severe anaemia in primigravidae were illiteracy and poor nutritional status (i.e. body mass index < 18.5 kg/m2 and mid upper-arm circumference < 23 cm). After forward, step-wise, regression analysis of relative risk (RR) factors and their 95% confidence intervals (CI), variables associated with an increased risk for moderately severe anaemia were iron deficiency (RR = 4.2; CI = 3.0-6.0) and malaria parasitaemia (RR = 1.9; CI = 1.3 2.7) in primigravidae, iron deficiency (RR = 4.1; CI = 2.7-6.3) and mid-upper-arm circumference < 23 cm (RR = 1.8; CI = 1.1-3.0) in secundigravidae, and iron deficiency in multigravidae (RR = 3.1; CI = 4.3-6.9). PMID- 10474638 TI - The impact of the local water-development programme on the abundance of the intermediate hosts of schistosomiasis in three villages of the Senegal River delta. AB - The populations of the potential intermediate hosts of Schistosoma mansoni and S. haematobium were studied for a year at transmission sites near three villages in the lower delta of the Senegal River. Biomphalaria pfeifferi, found to be widely distributed and increasingly abundant, appears to be well adapted to the new areas of irrigation (created by the dams at Diama and Manantali) thanks to its ability to withstand changes in temperature and to aestivate. This species is responsible for intense transmission of S. mansoni during the rainy season. In contrast, Bulinus globosus, the species responsible for the transmission of S. haematobium (which occurs during the dry season), had a more limited distribution. The changing distributions of these two snail species appear to be linked to changes in local ecology, themselves the result of the recent programme of water-development in the delta. PMID- 10474639 TI - Serological assays on eight cases of human dirofilariasis identified by morphology and DNA diagnostics. AB - Specimens of Dirofilaria sp. removed from eight Italian patients were identified as D. repens by morphology and confirmed as such by a PCR-based method of DNA analysis. Blood samples were also drawn from the patients so that two serological tests (ELISA and western blot), one based on the recognition of molecular markers recently identified in the somatic antigenic complex of D. repens, could be evaluated. The antigenic complex used in the ELISA only gave a weak sensitivity. However, the western-blot assays, based on the polypeptide molecular markers, were found to have greater sensitivity and should be useful in detecting human cases of dirofilariasis. PMID- 10474640 TI - Analysis of enterovirus sequences recovered from the cerebrospinal fluid of patients with epidemic neuropathy. AB - An outbreak of a previously unknown disease, termed epidemic neuropathy (EN), occurred in Cuba between 1991 and 1993. Although nutritional and oxidative stress in the population were rapidly associated with the disease, several findings were not compatible with such stress being the only cause. In the search for biological factors, samples of the cerebrospinal fluid (CSF) from patients with EN were studied and found to have a slowly progressing cytopathic effect (CPE) on VERO cells. Although the results of several studies indicate the presence of enteroviruses, the CPE and other physico-chemical characteristics are not typical of these viruses. Viral sequences have now been amplified from patients' CSF, using oligonucleotide primers homologous to the enterovirus 5' non-coding region. The sequences of the amplified region showed a high degree of variability (7% 69%) when compared with the coxsackievirus (Cox)A9 Griggs used as the reference strain. Furthermore, sequences differing by > 55% (58%-70%) were isolated from a single individual. These results indicate the generation in stressed individuals of enterovirus quasispecies with altered biological properties, and these could have played a major role in the neurological injury of EN. PMID- 10474641 TI - Human otoacariasis. AB - Accidental entry of insects and other arthropods is a common aetiology of aural foreign bodies (FB) presenting to accident and emergency departments. A retrospective study revealed that the FB in almost half (148) of 348 cases of aural FB investigated at the Universiti Sains Malaysia Hospital was an arthropod. The most common arthropod encountered was the cockroach, followed by a cattle tick. The high prevalence of tick infestation of the human ear canal (i.e. human otoacariasis) currently appears to be unique to the Malaysian state of Kelantan. The presentation of patients with intra-aural ticks, the methods used to remove the ticks, the complications encountered, and recommendations for an appropriate course of action in such cases are discussed. PMID- 10474642 TI - The influence of prematurity and low birthweight on transplacental antibody transfer in Sri Lanka. AB - The influence of gestational age, the neonate's birthweight, and maternal age, weight, height and parity on transplacental antibody transfer was assessed in 141 mothers from Sri Lanka and their neonates. Paired blood samples were collected from the mothers and the umbilical cords of the newborns. The sera separated from these samples were categorized as: preterm but adequate birthweight (< 37 weeks' gestation and birthweight > or = 2500 g); term but low birthweight (> or = 37 weeks' gestation and birthweight < 2500 g); or term and adequate birthweight (> or = 37 weeks' gestation and birthweight > or = 2500 g). Neonatal and maternal sera were assessed, in ELISA, for specific IgG antibodies against measles virus (MeV), herpes simplex virus type-1 (HSV1), respiratory syncytial virus (RSV), varicella-zoster virus (VZV), tetanus toxoid (TT), diphtheria toxoid (DT), and Streptococcus pneumoniae (Pn) and Haemophilus influenzae type-b (Hib) capsular antigens. Placental antibody transfer to certain antibody specificities was significantly lower in preterm neonates than term neonates. Thus the ratios between geometric mean cord antibody levels and geometric mean maternal antibody levels (the antibody-transfer ratios) were lower in preterm sera than term sera, for MeV (1.51 v. 2.03; P = 0.03), HSV1 (1.29 v. 1.76; P = 0.04), VZV (0.96 v. 2.50; P = 0.01), TT (1.13 v. 1.33; P = 0.04), DT (1.03 v. 2.39; P = 0.02), Pn (0.68 v. 0.98; P = 0.01) and Hib (0.58 v. 0.98; P = 0.00). Geometric mean levels of antibody to MeV, VZV, TT, DT and Pn were also significantly lower in preterm neonates than term. Compared with the values for 'adequate-birthweight' sera, low birthweight was independently associated with significantly lower levels of antibody transfer, for MeV (with antibody-transfer ratios of 1.51 v. 2.03; P = 0.02), VZV (0.99 v. 2.50; P = 0.03), TT (1.01 v. 1.33; P = 0.04) and DT (1.16 v. 2.39; P = 0.04) and significantly lower levels of antibodies to MeV, HSV1, VZV, TT, DT and Pn in the neonates. Maternal age, weight, height and parity had no independent influence on placental IgG transfer for antibodies to any of the pathogens investigated. These results demonstrate that prematurity and low birthweight may influence the level of maternally acquired immunity in Sri Lankan neonates. PMID- 10474643 TI - In-vitro activity of primaquine against the asexual blood stages of Plasmodium falciparum. PMID- 10474644 TI - Acute Chagasic cardiopathy in French Guiana: a re-emergent hazard linked to ecological change? PMID- 10474645 TI - Observations on leucine incorporation into sterol by Leishmania, and its inhibition by terbinafine. PMID- 10474646 TI - Acanthamoeba in human faeces from Karachi. PMID- 10474647 TI - Epidemiological study on the transmission of schistosomiasis in Saudi Arabia (western region). PMID- 10474648 TI - Histopathological and parasitological changes in baboons (Papio hamadryas) experimentally infected with baboon and human isolates of Schistosoma mansoni from Saudi Arabia: a comparative study. PMID- 10474649 TI - Serum levels of vascular cell adhesion molecule-1 (VCAM-1) in acute Q fever. PMID- 10474650 TI - Global warming and the performance of drugs used to treat parasitic and other diseases. PMID- 10474651 TI - A model for cytoplasmic rheology consistent with magnetic twisting cytometry. AB - Magnetic twisting cytometry is gaining wide applicability as a tool for the investigation of the rheological properties of cells and the mechanical properties of receptor-cytoskeletal interactions. Current technology involves the application and release of magnetically induced torques on small magnetic particles bound to or inside cells, with measurements of the resulting angular rotation of the particles. The properties of purely elastic or purely viscous materials can be determined by the angular strain and strain rate, respectively. However, the cytoskeleton and its linkage to cell surface receptors display elastic, viscous, and even plastic deformation, and the simultaneous characterization of these properties using only elastic or viscous models is internally inconsistent. Data interpretation is complicated by the fact that in current technology, the applied torques are not constant in time, but decrease as the particles rotate. This paper describes an internally consistent model consisting of a parallel viscoelastic element in series with a parallel viscoelastic element, and one approach to quantitative parameter evaluation. The unified model reproduces all essential features seen in data obtained from a wide variety of cell populations, and contains the pure elastic, viscoelastic, and viscous cases as subsets. PMID- 10474652 TI - Hyaluronan, hydration and flow conductivity of rat dermis. AB - We have measured the flow conductivity, kappa/eta, of discs of rat dermis with and without digestion with hyaluronidase. We found no significant difference between the flow conductivity of hyaluronan digested (kappa/eta = 5.56 +/- 2.74 cm4/dyne.s (n = 13)) and untreated tissue (kappa/eta = 6.03 +/- 3.15 cm4/dyne.s (n = 13)). For the first time in such experiments the overall tissue hyaluronan content as well as the difference in concentration of this material across tissues subjected to conditions of flow was measured. Similarly, the overall hydration and the difference in fluid content across the tissue is also reported. We have demonstrated that approximately 99% of the tissue hyaluronan was digested as a result of activity of the enzyme. No difference in hyaluronan across the tissue was found in the flow experiments (i.e., with or without digestion). We found significant change in the overall tissue hydration for controls and for either of the two types of flow experiments performed. Likewise, we found significant hydration differences across the tissues under both types of flow conditions. A trend in decreasing hydration associated with digestion of hyaluronan in the flow experiments was found. PMID- 10474653 TI - Shear stress gradient over endothelial cells in a curved microchannel system. AB - Our purpose was to test a scale model of the microcirculation by measuring the shear forces to which endothelial cells were exposed, and comparing this to computer simulations. In vitro experiments were performed to measure the 2 dimensional projected velocity profile along endothelial cell lined microchannels (D-shaped, 10-30 microns radius, n = 15), or in microchannels without endothelial cells (n = 18). Microchannels were perfused with fluorescently labeled microspheres (0.5 micron dia., < 1%) suspended in cell culture media. The velocity of individual microspheres was obtained off-line (videorecording), using an interactive software program; velocity was determined as the distance traveled in one video field (1/60 s). Mass balance was verified in the microchannels by comparing the microsphere velocities to the perfusion pump rate. In confluent endothelial cell lined microchannels, a velocity profile was obtained as microspheres passed an endothelial cell nucleus (identified by fluorescent dye), and again, for a paired region 100 microns away without nuclei (cytoplasm region). The velocity profile was significantly shifted and sharpened by the endothelial cell nucleus, as anticipated. Over the nucleus, data are consistent with a normal sized nucleus extending into the lumen, further confirming that this scale model can be used to determine the wall shear stress to which endothelial cells are exposed. Using the experimental bulk phase fluid parameters as boundary conditions, we used computational fluid dynamics (CFD) to predict the expected wall shear stress gradient along an endothelial cell lined D-shaped tube. The wall shear stress gradient over the nucleus was 2-fold greater in the radial versus axial directions, and was sensitive to lateral versus midline positioned nuclei. PMID- 10474654 TI - Wall shear stress in backward-facing step flow of a red blood cell suspension. AB - An experimental investigation of the wall shear stress distribution downstream of a backward-facing step is carried out. The wall shear stress distribution was determined by measuring the deformation of a gel layer, attached to the wall downstream of the step. Speckle pattern interferometry was applied to measure the deformation of the gel layer. The measured deformation, combined with the properties of the gel layer, served as an input for a finite element solid mechanics computation to determine the stress distribution in the gel layer. The wall shear stress, required to generate the measured deformation of the gel layer, was determined from these computations. A Newtonian buffer solution and a non-Newtonian red blood cell suspension were used as measuring fluids. The deformation of the gel layer was determined for a Newtonian buffer solution to evaluate the method and to obtain the properties of the gel layer. Subsequently, the wall shear stress distribution for the non-Newtonian red blood cell suspension was determined for three different flow rates. The inelastic non Newtonian Carreau-Yasuda model served as constitutive model for the red blood cell suspension. Using this model, the velocity and wall shear stress distribution were computed by means of a finite element fluid mechanics computation. From the comparison between the numerical and the experimental results, it can be concluded that wall shear stresses, induced by the red blood cell suspension, can be modeled accurately by employing a Carreau-Yasuda model. PMID- 10474655 TI - Low Reynolds number turbulence modeling of blood flow in arterial stenoses. AB - Moderate and severe arterial stenoses can produce highly disturbed flow regions with transitional and or turbulent flow characteristics. Neither laminar flow modeling nor standard two-equation models such as the kappa-epsilon turbulence ones are suitable for this kind of blood flow. In order to analyze the transitional or turbulent flow distal to an arterial stenosis, authors of this study have used the Wilcox low-Re turbulence model. Flow simulations were carried out on stenoses with 50, 75 and 86% reductions in cross-sectional area over a range of physiologically relevant Reynolds numbers. The results obtained with this low-Re turbulence model were compared with experimental measurements and with the results obtained by the standard kappa-epsilon model in terms of velocity profile, vortex length, wall shear stress, wall static pressure, and turbulence intensity. The comparisons show that results predicted by the low-Re model are in good agreement with the experimental measurements. This model accurately predicts the critical Reynolds number at which blood flow becomes transitional or turbulent distal an arterial stenosis. Most interestingly, over the Re range of laminar flow, the vortex length calculated with the low-Re model also closely matches the vortex length predicted by laminar flow modeling. In conclusion, the study strongly suggests that the proposed model is suitable for blood flow studies in certain areas of the arterial tree where both laminar and transitional/turbulent flows coexist. PMID- 10474656 TI - Swimming of spermatozoa in a linear viscoelastic fluid. AB - A modified resistive force theory is developed for a spermatozoon swimming in a general linear viscoelastic fluid. The theory is based on a Fourier decomposition of the flagellar velocity, which leads to solving the Stokes flow equations with a complex viscosity. We use a model spermatozoon with a spherical head which propagates small amplitude sinusoidal waves along its flagellum. Results are obtained for the velocity of propulsion and the rate of working for a free swimming spermatozoon and the thrust on a fixed spermatozoon. There is no change in propulsive velocity for a viscoelastic fluid compared to a Newtonian fluid. The rate of working does change however, decreasing with increasing elasticity of the fluid, for a Maxwell fluid. Thus the theory predicts that a spermatozoon can swim faster in a Maxwell fluid with the same expenditure of energy for a Newtonian fluid. PMID- 10474657 TI - Bioelectrorheological model of the cell. 8. Cellular deformation under prolonged and recurrent shear stress. AB - The influence of a prolonged and recurrent shear stress created by a periodic electric field on the mechanical properties of Neurospora crassa cells was investigated. Conditions were found under which modifications of cellular structures responding to stress become irreversible, and plastic flow of the viscoelastic structural elements is observed. The symmetry of the response of the cell under stress application and relaxation was lost, when compared to the reference conditions. To interpret the results a general rheological model was proposed. As previously described (Pawlowski et al., 1997), the existence of the three hypothetical supramolecular regions of the membrane (F, S and O) was suggested. Rheological parameters for the above regions were calculated. Theoretical functions were satisfactorily fitted to the experimental results. PMID- 10474658 TI - Complex viscoelasticity of normal and lectin treated erythrocytes using laser diffractometry. AB - A new method to find directly complex viscoelastic parameters (CVP) of red blood cells (RBC) is presented in this paper. Experimental determinations were carried out in an Erythrodeformeter (Rasia et al., 1986) operating in oscillating mode (0.5 to 3.5 Hz). The Erythrodeformeter performs direct determination of CVP of erythrocytes undergoing sinusoidal shear stresses by laser diffractometry. The measurements lead to the determination of mean values of the four components of erythrocyte complex viscoelasticity. The influence of the alterations induced on erythrocyte membrane by vegetable lactins (Ulex europaeus, wheat germ agglutinin and Enterolobium contorticilicum seeds) was analyzed to verify the sensitivity of this method. Differences observed between the CVP parameters of treated cells and the ones corresponding to control samples (non treated cells) are analyzed. Results obtained from cells treated with wheat germ agglutinin agree with observations published by Smith and Hochmuth (1982). Determinations of RBC complex viscoelasticity carried out by laser diffractometry could become an important tool to understand the influence of the factors associated with alterations of the rheologic properties of RBC membrane, which can affect the in vivo blood flow. PMID- 10474659 TI - Blood low shear rate rheometry: influence of fibrinogen level and hematocrit on slip and migrational effects. AB - Red blood cell (RBC) aggregation is of prime importance in vivo and in vitro for low flow rates. It may be estimated by rheometrical measurements at low shear rates, but these are perturbed by slip and migrational effects which have already been highlighted in the past. These effects lead to a torque decay with time so that the true value of the stress at low shear rates may be greatly underestimated. Elevated aggregation being associated with different diseases, pathological blood samples show more pronounced perturbing effects and a strong time dependency in low shear rate rheometry. To test the dependence of slip and migrational effects on RBC aggregation, and particularly to determine the way in which they depend upon fibrinogen concentration ([Fb]), a home-made measuring system with roughened internal and external walls (170 microns roughness) was used to study low shear rate rheometry for RBC suspensions in PBS buffer containing albumin (at 50 g/l) and fibrinogen at various concentrations. The influences of hematocrit, shear rate, and fibrinogen concentration were investigated. Particular attention was paid to data acquisition at low shear rates (10(-3) s-1 to 3 x 10(-2) s-1). The combined influence of hematocrit and fibrinogen was investigated by adjusting hematocrit to 44 or 57% and fibrinogen concentration ([Fb]) to 3.0-4.5-6.5 g/l. Microscopic observations of the blood samples at rest were performed. They showed that different structures were formed according to fibrinogen concentration. The rheometrical measurements indicated that torque decay with shearing duration was strongly dependent on fibrinogen concentration and on shear rate at fixed hematocrit. Migrational and slip effects were more pronounced as shear rate decreased, fibrinogen concentration was raised, and hematocrit was lowered. The results have been explained on the basis of the expected microstructure of flowing blood in relation to the microscopic observations at rest. PMID- 10474660 TI - Alterations in the biorheological features of some soft tissues after limb lengthening. AB - The aim of this study was to investigate the effects of a surgical limb lengthening procedure on the biorheological features of some lengthened soft tissues. In this procedure external fixators were applied to goats' right radius to stretch the tissues. The right forelegs of goats were lengthened by 2, 4 cm, respectively. After lengthening ceased, the goats were examined after different periods of time. The lengthened median nerves, arteries and veins were harvested and used to study their biorheological features. Tensile strength of lengthened and control specimens were measured and their stress relaxation features and stress-strain relationships were studied. Results showed that at the beginning of recovery, the stress-strain curves, relaxation curves and tensile strengths of the lengthened specimens began to deviate from those of their controls. However, with increasing recovery time, the curves and tensile strength of the lengthened specimens reverted to those of their controls. All the tissues studied exhibited the same behavior. PMID- 10474661 TI - A review of heterotopia and associated salivary gland neoplasms of the head and neck. AB - Salivary tissue neoplasms may involve normal, accessory and heterotopic salivary gland tissue. A case of Warthin's tumour originating from heterotopic salivary gland tissue of the upper neck is reported. The radioactive uptake of 131I, evidenced in the neck mass in its pre-diagnostic assessment, suggested a diagnosis of cervical node involvement from a primary malignant thyroid neoplasm. A critical review of the literature on heterotopic salivary gland tissue neoplasms of the head and neck is also presented. PMID- 10474662 TI - Alfred Gardiner Brown (1838-1882) M.R.C.S.Eng., F.R.C.S.(Ed.). His contribution to the history of the development of the tuning fork. AB - Alfred Gardiner Brown was the first designated Aural Surgeon to be appointed to the consultant staff of The London Hospital, at the age of 38. In July 1880 he published the paper showing his adaptation of the way in which the tuning fork was applied to the head and its use in measuring hearing power by comparing it with the sense of touch. Had he been blessed with a longer life it is highly probable that he would have made a significant contribution as he was of an innovative and inventive mind. He died at the age of 44. PMID- 10474663 TI - The acceptance of hearing aids for children with otitis media with effusion. AB - Conservative treatment for otitis media with effusion (OME) led us to consider the use of hearing aids as a way of managing the associated hearing loss. This study aimed to assess the compliance of patients and acceptance of hearing aids for the management of children with OME. Thirty-nine children who had been given binaural hearing aids to manage OME were assessed at routine follow-up after six months. A clinician who did not prescribe the aid administered a questionnaire to assess compliance, change in symptoms and acceptance of the aids. Thirty-eight parents thought the aids were easy to use and 25 (66 per cent) were completely satisfied with the management. Aided hearing improved by a mean of 17 dB (Range 10-30) over three frequencies, 0.5, 1.0, 2.0 Khz and all parents reported subjective hearing improvement in their children. The stigma of an aid was reported as minimal under the age of seven. Hearing aids provide a non-invasive way of managing the problems associated with OME which is acceptable to certain parents and children. Long-term effects of using aids need to be evaluated before they can be recommended. PMID- 10474664 TI - The influence of pregnancy on sensation of ear problems--ear problems associated with healthy pregnancy. AB - We wondered how many women had experienced a sensation of fullness in the ear during pregnancy. To address this question, data were obtained from a group of healthy women who attended the gynaecology clinic in our hospital as pregnancy cases between February 1995 and January 1998 and who volunteered to participate in our study. A control group was drawn from healthy female co-medical staff members of our hospital who had never been pregnant. The data used for comparing the two groups were taken from a questionnaire about ear problems that was presented to all subjects. The results suggest that ear problems may be increased in pregnancy, particularly for hypotensive pregnant women. However, even for pregnant women complaining of ear problems, pure-tone audiometry and impedance audiometry showed normal hearing in all cases. Furthermore, these women's complaints resolved completely on delivery of their babies. PMID- 10474666 TI - How uncomfortable are the various positions recommended for the instillation of nose drops? AB - Nose drops are widely used in the topical treatment of nasal disorders. Their efficacy has previously been shown to depend on the position of the head adopted during instillation. All three of the commonly recommended head positions (head back, praying-to-Mecca, Mygind's) are uncomfortable, and this may affect patient compliance. As yet, no assessment has been made of the discomfort encountered by patients. Twenty adults from a general otolaryngology clinic were asked to adopt each of three positions and then rate the discomfort experienced on a 10 cm visual analogue scale. The results show that although Mygind's position is well tolerated, the praying-to-Mecca position was significantly more uncomfortable than any other. In the absence of any evidence that the praying-to-Mecca position is clinically more efficacious than Mygind's, it seems inappropriate to continue to advocate its use. PMID- 10474665 TI - Clarion cochlear implants: surgical implications. AB - The surgical aspects of 34 Clarion cochlear implants, positioned during a five year period on 31 profoundly-deaf subjects as primary (11 children, 20 adults) and revision surgery (one child, three adults) were taken into consideration. Intra- and post-operative complications related to the specific structure of this device were considered along with the benefits obtained by update of the hardware and surgical approach. A smaller thickness of the internal receiver as well as the shifting from a retro-auricular/sub-temporal (RA-ST) to an endaural retromastoid (EA-RM) approach have contributed to a dramatic drop in post operative complications, such as skin defect or device extrusion. Special care should, however, be taken when cochlear implantation is planned in ears with chronic otitis or its surgical sequelae. From the two revised cases (hardware failure, electrode displacement) in whom re-implantation in the same side was performed, it has been demonstrated that the helicoidal shape of the electrode carrier is not inducing any additional trauma to the cochlear structure. PMID- 10474668 TI - A new method for evaluating and reporting the severity of snoring. AB - We propose a new questionnaire-based method for evaluating and reporting the severity of snoring, the snoring scale score (SSS), which provides an assessment of the loudness, frequency and periodicity of snoring sound. Ninety-nine patients with snoring problems were assessed and the results of 19 patients who underwent surgery to reduce snoring were reported using the SSS and the linear analogue score (LAS) at six weeks and six months after surgery. All patients reported reduction of snoring on both the SSS and LAS after surgery although minor discrepancies of reporting were found in six patients when comparisons were made between reporting of outcome using the SSS and LAS. Our study suggested that the SSS offers a uniform, comprehensive and simple to understand method for the evaluation, grading and reporting of snoring which is not currently available. PMID- 10474667 TI - Sequential magnetic resonance imaging analysis of the maxillary sinuses: implications for a model of gene therapy in cystic fibrosis. AB - Serial maxillary sinus aminoglycoside lavage is an adjunctive technique increasingly employed in a variety of areas in cystic fibrosis (CF). It may be helpful in reducing revision rates for sinus surgery, in lowering rates of bronchial pseudomonal colonization after lung transplantation and in the evolving field of gene therapy for CF. The goal of this study was to assess the utility of the maxillary sinus as a model for gene transfer in cystic fibrosis. We performed serial maxillary sinus lavage, in accordance with published protocols, using tobramycin in a randomized series of five CF subjects. Lavage was performed for up to 10 days and sequential magnetic resonance imaging (MRI) scans were taken at zero, 10, 30, 60, 120 and 180 days. The 30 MRI scans were blindly scored by two examiners on the parameters of maxillary sinus aeration, averaged over the five time intervals, was significantly improved (p < 0.05) in the lavaged sinus. This study provides the first systematic image-based measure of efficacy of maxillary sinus aminoglycoside lavage, a major element of a number of clinical protocols used in the treatment of CF. The prolonged increase in aeration after lavage suggests that any further improvement potentially achievable after gene transfer would be difficult to detect, limiting the value of this system as a model of clinical efficacy of gene transfer in CF. PMID- 10474669 TI - Effect of caffeine on the vocal folds: a pilot study. AB - Caffeine is considered to be a dehydrating agent with detrimental effects on the quality of voice of persons ingesting it. This has led medical personnel dealing with voice disorders, especially in the case of professional voice users, to give advice against the use of caffeine. Yet this is an anecdotal truth as an extensive Medline literature search did not reveal any scientific evidence of caffeine being proven to have adverse effects on the vocal folds. We, therefore, initiated this pilot study to ascertain the connection between caffeine and voice quality on a laboratory basis. Two hundred and fifty mg of caffeine were provided to eight volunteers in tablet form, and blood levels along with laryngograph readings were recorded to document the changes produced. Analysing the irregularities of frequencies in a) free speech b) a reading passage and c) singing 'Happy Birthday', substantial changes were seen to authenticate the fact that caffeine does produce alterations in voice quality but these alterations have considerable intra-subject variability. A full study with wider parameters is to be performed on this subject as we consider it to be of importance in the management of voice disorders. PMID- 10474670 TI - Lateral thyrotomy for neurilemmoma of the larynx. AB - We present a rare case of neurilemmoma of the larynx, which was excised through a lateral thyrotomy approach with resulting restoration of laryngeal function. The advantage of the lateral thyrotomy approach for submucous tumours of the larynx is discussed. PMID- 10474671 TI - Psychological morbidity following laryngectomy: a pilot study. AB - This study evaluated the nature and extent of problems faced by patients following discharge from hospital following laryngectomy. Semi-structured interviews investigated the practical and psychological concerns of a cohort of laryngectomy patients in a district general hospital. A number of simple practical measures can be employed to reduce early hospital re-admission following laryngectomy. Psychological morbidity is an important cause of low quality of life following laryngectomy. Formal psychological treatment could be of benefit in the management of these patients. PMID- 10474672 TI - Cochlear implantation of a deaf blind patient with mitochondrial cytopathy. AB - Genetic defects of the mitochondrial DNA often cause sensorineural hearing impairment, accompaniment by disorders of organs within the body. This case report describes cochlear implantation of a 33-year-old deaf blind female with mitochondrial cytopathy. The outcome was very successful, and vastly improved quality of life for this patient. Many cases of mitochondrial cytopathy cause progressive deafness; it is, therefore, likely that other patients with this unusual disorder will present for cochlear implant assessment. PMID- 10474674 TI - An unusual complication of augmentation rhinoplasty: late displacement of dorsal nasal splint. AB - Silastic (Dow-Corning) implants are used for augmentation rhinoplasty. Complications following their use usually occur within the first 24 months. We report a case in which the dorsal splint became displaced spontaneously five years after surgery. PMID- 10474673 TI - Metronidazole ototoxicity--report of two cases. AB - Two cases of bilateral moderate to severe sensorineural hearing loss due to oral administration of metronidazole are reported. There has been only one case report of deafness following metronidazole therapy in the world literature. The hearing loss recovered gradually in a period of four to six weeks following withdrawal of drug and oral steroid therapy. The possible mechanism of ototoxicity is discussed. Awareness by the treating physician of ototoxicity due to any drug is stressed. PMID- 10474675 TI - Laryngeal amyloidosis with laryngocele. AB - Both laryngocele and laryngeal amyloidosis are uncommon, and simultaneous occurrences of these entities are extremely rare. A case of laryngeal amyloidosis with laryngocele in which the computed tomography (CT) and magnetic resonance (MR) imaging of the larynx, clearly demonstrating both disease processes, is discussed. Diagnosis is confirmed by histopathologic specimens. Only two cases have been reported in the world literature, and this is the third case of laryngeal amyloidosis associated with laryngocele. PMID- 10474676 TI - Adult tracheocele with large cervical presentation. AB - A case is described in which a middle-aged male presented with a large tracheocele in the neck arising from the right postero-lateral aspect of trachea at the level of T2 vertebra. It was not associated with any predisposing factors and except for the swelling in the neck the patient was asymptomatic. It was excised completely through the cervical approach and there has been no recurrence in two years follow-up. PMID- 10474677 TI - Transpharyngeal approach for the treatment of dysphagia due to Forestier's disease. AB - Forestier's disease (diffuse idiopathic skeletal hyperostosis) is characterized by extensive spinal osteophyte formation and endo-chondral ossification of paravertebral ligaments and muscles. Dysphagia in the setting of Forestier's disease is a rare and hence often unrecognized entity. The dysphagia is due to mechanical obstruction in the initial stages and later due to inflammation and fibrosis. Most of these patients are treated conservatively in the initial stages and later by excision of osteophytes through a lateral cervical approach. We present a case of dysphagia due to cervical osteophytes in the setting of Forestier's disease causing narrowing of the pharynx. The patient was treated surgically via a peroral-transpharyngeal route with excellent results. PMID- 10474678 TI - An unusual presentation of an encephalocele to the otolaryngologist. AB - An unusual case of encephalocele causing obstruction of the upper airway in a neonate is described. The patient presented with a large mass in the neck, which extended from the base of the skull to the level of the larynx. Although there was no evidence of a defect of the skull base, subsequent excision and histological examination confirmed an encephalocele. PMID- 10474679 TI - Extracranial internal carotid artery aneurysm in a child: a diagnostic and surgical challenge. AB - This paper reports the presentation and management of an extra-cranial internal carotid artery aneurysm in a 15-year-old male. To our knowledge there is no previous report of a similar case in childhood. PMID- 10474680 TI - Primary osteogenic sarcoma of the tongue. AB - A 56-year-old man presented with the difficulty of swallowing and respiratory distress due to a large tumour arising from the tongue and occupying the entire oral cavity. Histological examination revealed it to be an extraskeletal osteogenic sarcoma. The tumour was excised. After six weeks, he came back with massive local recurrence and bleeding from the tumour, but died despite chemotherapy. Review of the literature revealed only four other such cases of this rare tumour. A brief review of these four cases is also made. PMID- 10474681 TI - Chondromyxoid fibroma of skull base: a tumour prone to local recurrence. AB - Chondromyxoid fibroma of the skull base is extremely uncommon. Sometimes involvement of the nasal cavity may occur and the patients may present with nasal symptoms. The biological behaviour of this tumour has not been well studied, primarily because of the limited number of reported cases and the short duration of follow-up. We report a histologically confirmed case of chondromyxoid fibroma of the skull base that recurred repeatedly over a 10-year period after the initial operation. Histologically it showed identical morphology to the original tumour with no evidence of histological progression or dedifferentiation. Ultrastructurally, the spindle tumour cells in the fibromyxoid area showed dual chondroblastic and fibroblastic differentiation, suggesting that these spindle fibroblastic cells and the better differentiated chondroid cells were of the same cell type with different histological morphology. PMID- 10474682 TI - Demystified ... oncogenes. PMID- 10474683 TI - Malignant fibrous histiocytomas and H-ras-1 oncogene point mutations. AB - AIMS: To investigate the types and the frequencies of H-ras-1 gene mutations in malignant fibrous histiocytomas. METHODS: Thirty five samples of malignant fibrous histiocytoma tissue were searched for point mutations within "hot spot" codons 12 and 13 of the H-ras-1 oncogene by the specific "nested" polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) and a direct cycle sequencing procedure. RESULTS: In contrast to previous reports, none of the tumours contained a point mutation or any other changes within or around the hot spot gene sequences. CONCLUSIONS: These data indicate that H-ras-1 oncogenic activation is not required in the molecular pathway of malignant fibrous histiocytoma formation and cannot be used as a discriminating factor for diagnostic sarcoma typing. PMID- 10474684 TI - Sensitive fluorescent in situ hybridisation method for the characterisation of breast cancer cells in bone marrow aspirates. AB - AIM: The presence of malignant cells in the blood and bone marrow of patients with cancer at the time of surgery may be indicative of early relapse. In addition to their numbers, the phenotypes of the micrometastatic cells might be essential in determining whether overt metastases will develop. This study aimed to establish a sensitive method for the detection and characterisation of malignant cells present in bone marrow. METHODS: In spiking experiments, SKBR3 cells were mixed with mononuclear cells in known proportions to mimic bone marrow samples with micrometastatic cells. Tumour cells were extracted using SAM-M450 Dynabeads coupled to the MOC-31 anti-epithelial antibody, and were further analysed for amplification of erbB2 and int2 by fluorescent in situ hybridisation (FISH). erbB2 and int2 copy numbers were also determined in 15 primary breast cancers, and bone marrow samples from patients with amplification were analysed for micrometastatic cells by immunomagnetic enrichment and FISH. RESULTS: In model experiments, cells with amplification could be detected in bead selected fractions when ratios of tumour cells (SKBR3) to mononuclear cells were as low as 10:10(7). Among the tumour samples, eight showed increased copy numbers of erbB2 and/or int2, and three of these patients had detectable numbers of tumour cells in their bone marrow: 4000, 540, and 26 tumour cells/10(7) mononuclear cells, respectively. The patient with 540 tumour cells/10(7) mononuclear cells showed high level amplification of erbB2 and suffered from a particularly aggressive disease, whereas the patient with 4000 tumour cells/10(7) mononuclear cells had favourable disease progression. CONCLUSION: These results demonstrate the feasibility and advantage of combining immunomagnetic selection and FISH characterisation of cancer cells in bone marrow samples. It is possible that molecular characterisation of such cells could provide prognostically valuable information. PMID- 10474685 TI - Fluorescence in situ hybridisation detection of erbB2 amplification in breast cancer fine needle aspirates. AB - AIM: To develop a method for the detection of amplification of the erbB2 oncogene in breast cancer fine needle aspirates using fluorescence in situ hybridisation (FISH) and to compare amplification with immunohistochemical detection of the erbB2 protein. METHODS: A digoxigenin labelled probe to the erbB2 gene was hybridised to 15 aspirates prepared from operative breast cancer specimens. A chromosome 17 centromere probe was also hybridised to the aspirates either separately or in combination with the erbB2 probe. The aspirates were scored for erbB2 amplification and chromosome 17 centromere number. Subsequently, paraffin wax embedded sections of the tumours were stained with the antibody CB11 and scored for the presence of membrane staining. RESULTS: Three of the 15 tumour aspirates showed high level amplification of erbB2 detected by FISH. These three tumours also showed chromosome 17 polysomy and diffuse membrane staining by immunohistochemistry. CONCLUSIONS: FISH can be used to detect erbB2 amplification in fine needle aspirates and results correlate with conventional immunohistochemical staining. Difficulties were encountered in the visualisation of the signals in non-amplified cases without the use of specialised digital imaging. PMID- 10474686 TI - Concerted overexpression of the genes encoding p21 and cyclin D1 is associated with growth inhibition and differentiation in various carcinomas. AB - AIMS: To investigate the expression of the genes encoding cyclin D1 and p21 in proliferative and non-proliferative cells, as demonstrated by the Ki67 antibody, and to correlate these findings with differentiation. METHODS: Immunohistochemistry and immunofluorescence double staining were performed on three breast cancers, two squamous cell cancers of the head and neck, and one ovarium cystadenocarcinoma. In addition, the in vitro effect of cyclin D1 on p21 gene expression in MCF7 breast cancer cells was evaluated. RESULTS: Immunofluorescence double staining showed a differentiation related gradient in the detection of the Ki67 antigen, cyclin D1, and p21 in squamous cell cancers of the head and neck: Ki67 was detected in the basal layers of the tumour and the cyclin D1 and p21 genes were coexpressed in the higher, more differentiated layers of the tumour. The breast and ovarian cancers often had cells that coexpressed the p21 and cyclin D1 genes, whereas coexpression of cyclin D1 and Ki67 did not occur. Western blot analysis of the MCF7 breast cancer cells showed an upregulation of p21 production when cyclin D1 gene expression was induced. CONCLUSION: Overexpression of the cyclin D1 gene seems to lead to growth arrest in a variety of human cancers, possibly through the induction of p21 by cyclin D1. In squamous cell cancer, concerted overexpression of the genes encoding cyclin D1 and p21 might also induce differentiation. PMID- 10474687 TI - Nuclear localisation of NOVH protein: a potential role for NOV in the regulation of gene expression. AB - AIMS: To identify the NOV protein detected by immunofluorescence in the nucleus of human cancer cell lines to establish whether targeting to the nucleus reflects dual paracrine and intracrine biological functions of NOV, as has been reported previously for several signalling peptides and proteins. METHODS: Nuclear and cytoplasmic fractions were prepared from 143 and HeLa cells in which nuclear NOV protein was detected. Western blotting analysis of NOV proteins in both types of fractions was performed using two NOV specific antibodies. Confocal microscopy was used to visualise the nuclear NOV protein in HeLa and 143 cells. A yeast two hybrid screening system was used to isolate cDNAs encoding proteins able to interact with the human NOV protein. RESULTS: A 31/32 kDa doublet of NOV protein was identified in the nuclear fraction of 143 and HeLa cells. Because the antibodies were directed against the C-terminus of NOV, the 31/32 kDa NOV isoform is probably truncated at the N-terminus and might correspond to the secreted 32 kDa NOV isoform detected in cell culture medium. Confocal microscopy indicated that in addition to the cytoplasmic NOV protein already identified, a nuclear NOV protein was present in both the nucleoplasm and nucleoli of Hela and 143 cells. Screening of cDNA libraries prepared from HeLa cells, Epstein-Barr virus transformed lymphocytes, and normal human brain showed that the NOV protein interacts with the rpb7 subunit of RNA polymerase in a yeast two hybrid system. CONCLUSIONS: The NOV protein detected in the nucleus of 143 and HeLa cells is probably an N-terminus truncated isoform of the secreted 48 kDa NOV protein. A growing body of evidence suggests that novH expression is closely associated with differentiation in normal human tissues and that the nov gene encodes a signalling protein that belongs to an emerging family of cell growth regulators. The nuclear localisation of a NOV isoform potentially provides an additional degree of signalling specificity. The interaction of the NOV protein and the rpb7 subunit of RNA polymerase II in the two hybrid system suggests that NOV might be involved in regulating gene expression at the transcriptional level. As has already been suggested for several other nuclearly located cytokines, the NOV protein does not contain a typical nuclear localisation signal. Therefore, it is possible that it combines with either a receptor or a chaperone during its translocation. Disruption of the balance between the secreted and nuclear NOV isoforms might affect the putative autocrine and paracrine functions of NOV and might be of considerable importance in the development of cancers in which the expression of novH has been shown to be impaired. PMID- 10474688 TI - Expression of the gene encoding the matrix gla protein by mature osteoblasts in human fracture non-unions. AB - BACKGROUND: Osteoblast phenotypic abnormality, namely the expression of collagen type III, has been shown previously in fracture non-union woven bone. AIMS: To investigate osteoblasts from fracture non-unions for evidence of gene expression of non-collagenous bone matrix proteins that have been implicated in mineralisation, namely matrix gla protein (MGP), osteonectin, osteopontin, and osteocalcin. MGP is a consistent component of bone matrix, but there are no reports of osteoblasts in the skeleton expressing the gene for MGP, and the site of synthesis of skeletal MGP (perhaps the liver) has yet to be determined. METHODS: Biopsies from normally healing human fractures and non-unions were examined by means of in situ hybridisation, using 35S labelled probes and autoradiography to disclose levels of gene expression. RESULTS: In normally healing fractures, mature osteoblasts on woven bone were negative for MGP mRNA, but positive for osteonectin, osteopontin, and osteocalcin mRNA molecules. In non unions, osteoblasts displayed a novel phenotype: they were positive for MGP mRNA, in addition to osteonectin, osteopontin, and osteocalcin mRNA molecules. CONCLUSIONS: Mature osteoblasts in slowly healing fractures have an unusual phenotype: they express the gene encoding MGP, which indicates that control of osteoblast gene expression in non-unions is likely to be abnormal. This might be of importance in the pathogenesis of non-uniting human fractures, and is of current interest given the emerging status of MGP as an inhibitor of mineralisation. PMID- 10474689 TI - Expression of Epstein-Barr virus (EBV) transcripts encoding homologues to important human proteins in diverse EBV associated diseases. AB - AIMS: To examine the expression of Epstein-Barr virus (EBV) transcripts encoding proteins homologous to important human proteins in diverse EBV associated diseases. The proteins were: BHRF1 (homologous to Bcl-2), BDLF2 (homologous to cyclin B1), BARF1 (homologous to intercellular cell adhesion molecule 1 (ICAM 1)), and BCRF1 (viral IL-10 (vIL-10), homologous to human IL-10 (hIL-10)). METHODS: Six cases of oral hairy leukoplakia, seven of Hodgkin's disease, eight of T cell non-Hodgkin's lymphoma, and nine of nasopharyngeal carcinoma were examined at the mRNA level using either the reverse transcriptase polymerase chain reaction (RT-PCR) or nucleic acid sequence based amplification (NASBA). Different primer sets allowed the differentiation by RT-PCR of the latent (Cp/Wp driven) and lytic (Hp driven) transcripts of BHRF1. A specific NASBA reaction was developed for the detection of vIL-10 and BDLF2 transcripts and this was tested initially on cell lines and later on clinical samples. RESULTS: vIL-10 and BDLF2 were expressed almost exclusively in oral hairy leukoplakia, whereas BARF1 transcripts were present in all cases of nasopharyngeal carcinoma, with weak expression in one oral hairy leukoplakia and isolated cases of lymphoid malignancy. Both BHRF1 transcripts were detected across the range of tissues tested, but strong expression of lytic BHRF1 transcripts was seen only in oral hairy leukoplakia. CONCLUSIONS: vIL-10 and BDLF2 transcripts are expressed during productive EBV infection and are unlikely to be important in the pathogenesis of EBV associated malignancies. BARF1 appears to be expressed preferentially during viral latency and is more closely associated with malignant rather than benign epithelial proliferations. The alternative transcripts derived from the BHRF1 open reading frame may have very different roles during latent or productive infection. PMID- 10474691 TI - Lessons learned and not learned from the SUPPORT project. PMID- 10474690 TI - Detection of clonal B cells in microdissected reactive lymphoproliferations: possible diagnostic pitfalls in PCR analysis of immunoglobulin heavy chain gene rearrangement. AB - AIMS: To evaluate the specificity of standard and fluorescence based (Genescan) polymerase chain reaction (PCR) immunoglobulin heavy chain (IgH) gene rearrangement analysis in complete and microdissected paraffin wax embedded sections from lymphoid proliferations. METHODS: PCR IgH gene rearrangement analysis of whole sections and microdissected fragments (n = 62) from paraffin wax embedded reactive lymph nodes (n = 6) and tonsils (n = 3). Amplificant analysis used both standard methods and automated high resolution fluorescence based quantification and size determination using GENESCAN software. RESULTS: Whole tissue sections were consistently polyclonal in control experiments. IgH gene amplification was successful in 59 of 62 microdissected fragments; only two of 59 showed a polyclonal rearrangement pattern, the remainder being oligoclonal or monoclonal. Reanalysis was possible in 33 samples; six showed reproducible bands on gel analysis and satisfied accepted criteria for monoclonality. Use of high resolution gels with Genescan analysis improved sensitivity and band definition; however, three samples still appeared to be monoclonal. CONCLUSIONS: These results confirm that PCR based IgH gene rearrangement analysis is a sensitive and specific method for demonstrating B cell clonality in whole paraffin wax embedded sections. However, oligoclonal and monoclonal rearrangement patterns are regularly encountered in small tissue fragments from otherwise unremarkable reactive lymphoproliferations, possibly because of preferential priming or detection of local B cell clones. Data from clonal analysis of small, microdissected or lymphocyte poor samples must be evaluated critically. It is recommended that analyses should be run in parallel on at least two tissue specimens. Only reproducible bands present in more than one sample should be considered to be suggestive of neoplasia. PMID- 10474692 TI - The role of morphine glucuronides in cancer pain. AB - Morphine metabolites are involved in various ways in determining the complex effects of morphine, both favourable and adverse, and may complicate the clinical use of morphine in the treatment of cancer pain. The production and effects of the principal morphine metabolites, morphine-3-glucuronide and morphine-6 glucuronide, in both normal and pathological states have been reviewed in the current literature. Therapeutic implications are also reviewed on the basis of experimental and clinical reports. The presence of these metabolites should be recognized in the chronic treatment of cancer pain with morphine, especially in the presence of renal impairment, and should be considered to have an important influence on opioid responsiveness, defined as a balance between the achievement of an optimal analgesia and the occurrence of adverse effects. PMID- 10474693 TI - Descriptive epidemiological survey on a given day in all palliative care patients hospitalized in a French university hospital. AB - We conducted a descriptive study on a given day on all inpatients requiring palliative care in a French university hospital. In each department, a collaborative team made up of physicians and nurses identified and described the clinical signs, the treatment protocols, the social and family characteristics and the outcome for each patient using a standardized questionnaire. Study subjects were inpatients in the hospital and presented advanced or terminal-stage life-threatening conditions. Two-hundred-and-forty-five patients were included in the study. These patients represented 13% of the total number of inpatient beds available in the hospital on the day of the survey. Sixty-six per cent of study subjects suffered from physical discomfort and 80% suffered psychologically. Patients still received specific treatment for their condition in 45% of cases. Social problems were identified principally in medium- or long-term care department inpatients who made up 36% of the total inpatient population. A request for transfer to another care structure had been completed for 24% of patients. Assistance from the Palliative Care Unit's support team had been requested in 25% of cases, mainly to provide psychological support for the patient and the health care providers. These results have led us to reconsider the general organization of palliative care in the health care system. PMID- 10474694 TI - Community-based palliative care for Bangladeshi patients in east London. Accounts of bereaved carers. AB - The aim of this paper is to describe the palliative care experience of Bangladeshi patients and carers in the Tower Hamlets area in the east of London. Semi-structured interviews were carried out in Sylheti, the Bengali dialect of this community, with bereaved primary carers of 18 patients (10 male, eight female) referred to an east London community palliative care team between 1986 and 1993. It was found that patients were young, with a mean age of males of 55 years (range 34-65) and females of 40 years (range 28-57). Communication difficulties were common. The fluency in English of patients was low, with reliance on family members, especially children, for translation. The diagnosis was known by all patients, but only 56% of carers agreed with disclosure. Team dissatisfaction with communication was recorded in 16 cases. Fourteen patients died in London; however, 13 were buried in Bangladesh. Carers often reported symptoms as poorly controlled. Pain was said to be severe for 14 patients, and pain control said to be poor in 11. Family and friends provided most support during the illness and bereavement. Serious financial difficulties occurred in nine families. General practitioners were involved actively in six cases. In conclusion, there are ethno-specific needs in this particular community, many of which arise from socio-economic factors, recent migration and religious beliefs, and which are highlighted by terminal illness. PMID- 10474695 TI - Improving palliative care services: a pragmatic model for evaluating services and assessing unmet need. AB - Evaluating local palliative care services and identifying gaps in services for patients are crucial to the development of services which enable people to die at home in a well-supported environment. A review of the local strategy for providing respite and support services for palliative care patients and their carers was carried out in one area of south-east England. The review comprised four elements: identification of services available; interviews with provider 'stakeholders'; interviews with patients and carers; questionnaire survey of general practitioners and district nurses. The findings and outcome are described. It is suggested that this approach could be adopted by health authorities to form one part of a comprehensive system of needs assessment. Alternatively, it could be used by groups of providers to evaluate existing services and to identify potential improvements to services. Advantages and disadvantages of the approach for both commissioners of such reviews and those carrying them out are discussed. PMID- 10474696 TI - 'Someone to talk to' and 'pain control': what people expect from a specialist palliative care team. AB - This paper presents a qualitative analysis of a subsample of data from a larger investigation into general practitioner referrals to specialist palliative care services (SPCS). The aim was to identify the expectations and perceptions of patients with advanced cancer and their relatives. Twenty-nine individuals were interviewed (18 patients, 11 relatives). Qualitative analysis of the interview transcripts was undertaken using an approach based on interpretative phenomenological analysis. The results indicated that SPCS were perceived as providing psychosocial services, characterized as 'support' and 'someone to talk to'. SPCS were expected to have 'expertise in symptom control', particularly in relation to pain management. The study demonstrated that patients' and relatives' expectations of a referral to SPCS were of psychosocial support and symptom control. It is argued that the implications of this restricted view of SPCS may mean that other salient concerns are not raised by patients and relatives. PMID- 10474697 TI - Unlicensed uses for medication in a palliative care unit. AB - The aim of this study was to review the extent of drug use for unlicensed purposes in a palliative care unit. We carried out a prospective study of all patients with advanced malignancy admitted to a 10-bed specialist palliative care unit over a 4-month period. Prescriptions were assessed and compared with licensed prescribing indications. Seventy-six patients were included in the study; 689 prescriptions were made using 84 drugs to treat 34 different symptoms. Fifteen per cent of prescribing events were for unlicensed indications. We conclude that drugs are frequently used in the palliative care setting for purposes unsupported by product licences, although usually backed by literature. These drugs are often prescribed for symptoms which are difficult to control. PMID- 10474698 TI - The use of interpleural analgesia using bupivacaine for pain relief in advanced cancer. AB - The use of interpleural analgesia is described in six patients with a variety of advanced malignancies suffering from pain uncontrolled by opioids. The benefits and complications of the technique are discussed including management of the catheters at home and the measurement of plasma bupivacaine concentrations. Interpleural analgesia can provide good analgesia in a small, selected population of patients with otherwise uncontrolled pain of malignant origin. PMID- 10474701 TI - A year of acupuncture in palliative care. PMID- 10474700 TI - Ethnic minorities and palliative care in Derby. PMID- 10474699 TI - Morphine, constipation and performance status in advanced cancer patients. PMID- 10474702 TI - Percutaneous endoscopic duodenostomy: the relief of obstruction in advanced gastric carcinoma. AB - Nausea and vomiting in patients with advanced gastric malignancy and mechanical obstruction are distressing and difficult to manage. We describe a patient with linitis plastica and gastric stasis who was treated with a percutaneous endoscopic duodenostomy as the stomach could not be used for percutaneous endoscopic gastrostomy (PEG) formation. A Conflo PEG tube was inserted into the second part of the duodenum using the Ponsky-Gauderer technique without complication. The patient experienced excellent symptomatic relief and tolerated enteral nutrition extremely well, regaining some weight. This manoeuvre can produce effective symptom palliation allowing the patient to be managed at home during the terminal phase of their illness. PMID- 10474703 TI - How do patients who die at home differ from those who die elsewhere? PMID- 10474705 TI - Specialist palliative care needs specialists. PMID- 10474704 TI - Alternative treatments at the end of life. PMID- 10474706 TI - Nonsteroidal anti-inflammatory drugs as adjuvant analgesics in cancer patients. AB - The role of nonsteroidal anti-inflammatory drugs (NSAIDs) is examined in the control of cancer pain with a particular focus on their use as adjuvants to opioids in advanced cancer pain. These agents have both a peripheral effect on inflammation and a role in attenuating central pain pathways. The possibility of obtaining the benefits of NSAIDs with fewer side-effects by using COX-2-specific agents is discussed. The gastrointestinal, renal, haemostatic, cognitive and hypersensitivity side-effects of NSAIDs are reviewed and their potential impact assessed. The evidence for the efficacy of NSAIDs as single agents for cancer pain is reviewed together with the nine papers which have reported the effects of NSAIDs as adjuvants to opioids in cancer pain. All of these papers reported positive results of NSAIDs, but, in the absence of any randomized, double-blind controlled trials, where NSAIDs were used as adjuvants on a long-term basis alongside optimal opioid use, definite conclusions cannot be reached. Guidelines for the safe use of NSAIDs are suggested. Finally, suggestions for future research are made. PMID- 10474707 TI - Patient and carer preference for, and satisfaction with, specialist models of palliative care: a systematic literature review. AB - This systematic literature review examined the impact of specialist models of palliative care on consumer satisfaction, opinion and preference over the past 20 years. From the literature search, 83 relevant papers were identified. Research findings from North America did not reveal any reliable or consistent trends, and this was due primarily to methodological flaws in the research. In the UK, consumers are more satisfied with all types of palliative care, whether provided by inpatient units or in the community, than with palliative care provided by general hospitals. Even though research findings consistently indicate that consumers appreciate the psychosocial climate in hospices, this research was based on small-scale local studies which were mainly focused on a single hospice. The dearth of high quality, comprehensive research was notable on the impact on consumer preference, opinion or satisfaction of hospice home care services, and other forms of palliative care in the community. It was concluded that further research is needed into consumer priorities for patients dying of noncancer diseases, and into the possible impact of patient characteristics on consumer satisfaction, opinion and preference for specialist models of palliative care. PMID- 10474708 TI - Encounters with doctors: quantity and quality. AB - Continuity of care and the large numbers of health care professionals who deliver that care are issues that frequently concern patients and their families. This study examined the number of doctors encountered by 50 patients, during the period of their cancer care. This ranged from 4 months to 26 years, with a median time of 2 years and 4 months. The doctors included in this number were general practitioners, doctors met during hospital inpatient admissions and when attending outpatient appointments, and doctors at the hospice. Descriptive statistics are included detailing the total number of doctors encountered by patients; the number met by patients within the first year of their cancer care; and the average number of new doctors met each year. The minimum number of doctors met was 13, maximum 97 and median 32. Notable examples include one patient who met 31 doctors during a 6-month period, and one patient who met 73 doctors during a period of 2 years and 1 month. Patients in this study with a history of less than 1 year met 28 doctors on average. Semi-structured interviews with these patients were conducted adopting a qualitative approach. Patients were asked about their recollections of the doctors they had met during their cancer care and what value they attributed to these encounters. Interviews were subject to thematic analysis. The major themes to emerge were: continuity of care, the provision of information and explanations and honesty in that process, breaking of bad news, the manner adopted by the doctor and issues relating to specialist referral. The large number of health care professionals, including the doctors quantified in this study, involved in the care of each patient represents a major challenge to 'seamless' and consistent communication between those involved. PMID- 10474709 TI - A survey of nursing and medical staff views on the use of cardiopulmonary resuscitation in the hospice. AB - Research evidence suggests that cardiopulmonary resuscitation (CPR) would be indicated in very few hospice patients. However, with the increasing access and expansion of specialist palliative care services the question of CPR is becoming more important. In order to develop a policy in our unit we felt it was important to assess the understanding, attitudes and experience of the health care professionals involved. A semi-structured questionnaire regarding CPR issues, including case scenarios, was distributed to doctors and registered nurses in a palliative care unit. Thirty-seven (80%) of the questionnaires were returned. Ten per cent of respondents identified patients for whom they felt CPR would have been indicated in the event of an unexpected cardiac arrest. Thirty-two per cent could foresee the number of patients in this category increasing in the future. The majority of respondents indicated that CPR should be discussed in certain cases, however 86% had never done so. The success rate of CPR was frequently overestimated. Some respondents felt vulnerable as there was no existing written policy. Factors thought important in making decisions regarding CPR orders included: prognosis; patient's wishes; quality of life; and legal issues. CPR in palliative care units raises many practical and ethical concerns. Our survey shows that staff are aware of the small, but increasing, need for its consideration in certain cases. There was a wide range of views regarding the role of CPR with an overestimation of the chances of success and concerns regarding discussion of the issue with patients. When introducing a CPR policy in a palliative care unit, adequate education and a framework for decision making is required. PMID- 10474710 TI - Home palliative care for terminal cancer patients: a survey on the final week of life. AB - As part of a large multicentre study on palliative care units in Italy, carried out between 1 January and 30 June 1995, we describe the place, circumstances and 'quality of death' of patients admitted to home palliative care. Data presented refer to 401 patients (67% of the 601 patients randomly selected for evaluation). Of these 401 patients 303 (76%) died at home. According to the Support Team Assessment Schedule (STAS) pain was fairly well controlled during the final week of life, while the control of other symptoms appeared to be less satisfactory. Invasive procedures were undertaken on 56% of patients, while in hospital the percentage increased to 75%. Twenty-five per cent of patients were totally pharmacologically sedated during the final 12 h of life. Neither the number of symptoms nor other factors were apparently associated with the decision to sedate the patient. The wide variations in the frequency of sedation among centres suggest that the choice to sedate the patient may reflect the provider's behaviour or services' policy rather than the patients' preference or needs. The definition of common criteria and guidelines for sedation of patients should be one of the topics for discussion among palliative care teams. PMID- 10474711 TI - A survey of antidepressant prescribing in the terminally ill. AB - Depression is a symptom in a quarter of patients admitted to a palliative care unit, but little is known of how depression in terminally ill patients is treated. We reviewed 1046 consecutive patient admissions, of whom 106 (10%) were prescribed antidepressant medication while under the care of a palliative care team. Of these patients, 21 were prescribed antidepressants when under the care of the home care team, but 80 patients (76%) were started on medication during the final 2 weeks of life. There was consequently insufficient time for the medication to have any therapeutic effect. Seventeen patients were discharged home on antidepressant medication. Three patients were referred for a psychiatric assessment. Patients prescribed antidepressants were significantly younger (P = 0.002) than those who were not. There were no prescriptions for psychostimulants. Although the numbers of patients prescribed antidepressant medication were low in all disease groups, it was notable that patients with breast cancer were prescribed antidepressant medication more frequently than any other patient group. We conclude that there appears to be a need for a coordinated approach to both the assessment and the treatment of depression in terminally ill patients. PMID- 10474712 TI - Joint surgical/palliative care ward round in a district general hospital. PMID- 10474713 TI - Treatment of odynophagia and dysphagia in advanced cancer with sublingual glyceryl trinitrate. PMID- 10474714 TI - Auricular stud acupuncture in palliative care patients. PMID- 10474715 TI - The 'n = 1' randomized controlled trial. PMID- 10474716 TI - Improvement of transurethral catheterization in male patients. PMID- 10474717 TI - The decision-making process in sedation for symptom control in Japan. PMID- 10474718 TI - The use of amitriptyline in mycosis fungoides. PMID- 10474719 TI - What's happening in undergraduate medical education? PMID- 10474720 TI - HIV infection and seizures. AB - New-onset seizures are frequent manifestations of central nervous system disorders in patients infected with human immunodeficiency virus (HIV). Seizures are more common in advanced stages of the disease, although they may occur early in the course of illness. In the majority of patients, seizures are of the generalised type. Status epilepticus is also frequent. Associated metabolic abnormalities increase the risk for status epilepticus. Cerebral mass lesions, cryptococcal meningitis, and HIV-encephalopathy are common causes of seizures. Phenytoin is the most commonly prescribed anticonvulsant in this situation, although several patients may experience hypersensitivity reactions. The prognosis of seizure disorders in HIV-infected patients depends upon the underlying cause. PMID- 10474722 TI - Questionnaires: the use and abuse of social survey methods in medical research. AB - We present a working review of survey methods based on market research technology. The structure of questionnaires, their distribution and analysis, are considered, together with techniques for increasing response rates. PMID- 10474723 TI - Induction for senior house officers. Part II: The departmental programme. AB - This study was designed to examine the content and usefulness of departmental induction programmes to senior house officers (SHOs) and to explore perceptions of the usefulness of a range of topics to these trainees. A total of 64 SHOs, in five hospitals in the Anglia region, participated in semi-structured interviews up to 3 months after starting their post. The interviews investigated the content of any induction received and also examined what the trainees would have liked to have received. Almost half (29) of the SHOs also completed a questionnaire which examined the perceived usefulness of various induction topics for a trainee starting a new post. The results showed that, although a departmental induction programme is considered important and highly valued, a substantial minority of SHOs had not received one. Of this minority, 75% would have welcomed an induction. Where an induction had taken place, the focus was primarily upon timetables, tours and meeting people, however, a quarter of the questionnaire sample had not received a service timetable, a third had not met any consultants, two-thirds had received no introduction to clinical management, and two-thirds had received no information about how their consultants manage their patients. Thus, although many trainees receive an induction, important information may not be covered. A staged approach to departmental induction is recommended, using a short, high-quality, and comprehensive induction programme. Flexible and timely programmes, supported by comprehensive written information, will contribute to meeting the needs of trainees in an efficient and effective way. PMID- 10474721 TI - The pathophysiological and molecular basis of Bartter's and Gitelman's syndromes. AB - Molecular defects affecting the transport of sodium, potassium and chloride in the nephron through the ROMK K+ channel, Na+/K+/2Cl- cotransporter, the Na+/Cl- cotransporter and chloride channel have been identified in patients with Bartter's and Gitelman's syndromes. Defects of the angiotensin II type I receptor and CFTR have also being described. These defects are simple (i.e., most are single amino acid substitutions) but affect key elements in tubular transport. The simplicity of the genetic defects may explain why the inheritance of these conditions remains unclear in most kindreds (i.e., not just recessive or dominant) and emphasises the crucial importance of the conformational structure of these channels. Application of this molecular information will allow the early genetic identification of patients with these syndromes and enable us to differentiate between the various disorders at a functional level. It may also identify a subgroup in which the heterozygous form may make patients potentially exquisitely sensitive to diuretics. PMID- 10474724 TI - Achieving a patient-centred consultation by giving feedback in its early phases. AB - The traditional medical consultation comprises history, examination, and investigations, followed by explanation to the patient of diagnosis and management. In the course of studying a series of tape-recorded consultations in a specialist medical clinic for chronic fatigue, we have observed a different structure. In some consultations, those categorized as more 'patient-centred', doctors introduced explanation and education into the early history-taking stage. This strategy is contrasted with the traditional approach, where the doctor only elicits information during the history, and gives an explanation later. The 'early feedback' strategy may result in patients with chronic illnesses achieving greater understanding of their symptoms. We discuss the implication of these findings for medical training. PMID- 10474725 TI - Sexual harassment of psychiatric trainees: experiences and attitudes. AB - A survey was carried out of psychiatric trainees' work-related experiences of unwanted sexual contact. A structured postal questionnaire was administered to 100 psychiatric trainees from senior house officer to specialist registrar level in a large psychiatric rotation. There was an 85% response rate; 86% (73) of the sample had experienced unwanted sexual contact, with 47% (40) experiencing deliberate touching, leaning over or cornering, and 18% (15) receiving letters, telephone calls or material of a sexual nature. Three-quarters (64) of respondents had experienced unwanted sexual contact from patients and 64% (54) from staff. Experiences and attitudes did not generally differ by gender, grade or training experience. Four out of 48 female respondents described stalking by patients. Of the 39 respondents who had reported harassment by patients, 31 felt supported by colleagues, while of the 13 who had reported harassment by colleagues, eight felt supported. Two-thirds of the respondents considered sexual harassment 'some-times' or 'frequently' a problem for the profession. Diagnoses of confusional states, mania or schizophrenia made subjects less likely to consider unwanted sexual behaviour to be 'sexual harassment' (86%, 80%, and 67%, respectively), but not for other diagnoses. Levels of threatening and intrusive sexual harassment are unacceptably high in this study group. Health trusts should adopt policies of 'zero tolerance' and all incidents should be reported. Psychological impact on victims should be acknowledged even when the behaviour of the perpetrator can be explained by diagnosis. PMID- 10474726 TI - Prospective audit of a respiratory sleep disorders service at District General Hospital level. AB - This study was designed to examine the organisation and outcomes of a District General Hospital respiratory sleep service, since data are lacking on the management of sleep-disordered breathing at this level. Questionnaires and case notes review were used to assess the management of 119 consecutive patients referred with suspected sleep-disordered breathing. Patients diagnosed with sleep disordered breathing were assigned nasal continuous positive airway pressure (nCPAP), ear/nose/throat (ENT) surgery or simple measures (e.g., weight loss). There were six non-attenders. At 12 months follow-up, 33 patients had been assigned to nCPAP, 25 to ENT surgery, and 37 to simple measures. Of the remainder, nine had alternative diagnoses, two were still being assessed and seven were lost to follow-up. Patients prescribed nCPAP (81% compliance) had significant symptomatic improvements with low dissatisfaction rates (20%); patients on simple measures did not improve (33% dissatisfied); only half assigned surgery had it performed, with 42% awaiting surgery and dissatisfied. Interspecialty referral resulted in major delays (mean 16 weeks). Referral letters were generally unhelpful in deciding on the appropriateness of initial referral (respiratory physician vs ENT). nCPAP was generally effective in improving symptoms, with a high level of patient satisfaction, while simple measures did not improve symptoms and were associated with lower satisfaction levels. Waiting times to ENT surgery can be long and patients express significant dissatisfaction. Referral letters are not useful in directing initial referral. Services should be co-ordinated between respiratory and ENT specialties to reduce waiting times and improve patient satisfaction. PMID- 10474727 TI - The hibernating heart: reversible left ventricular dysfunction in chronic heart failure. AB - A patient with severe heart failure secondary to coronary heart disease is presented. Following investigation he was thought to have significant areas of myocardial hibernation and was therefore treated with coronary revascularisation, with major clinical benefit. PMID- 10474728 TI - Clearance of acanthosis nigricans associated with the HAIR-AN syndrome after partial pancreatectomy: an 11-year follow-up. AB - We describe a woman with the syndrome characterised by hyperandrogenism, insulin resistance and acanthosis nigricans (the HAIR-AN syndrome), and an associated insulinoma (islet B-cell tumour), whose signs and symptoms cleared after partial pancreatectomy. PMID- 10474729 TI - Conjunctival MALT lymphoma: an usual cause of red eye. AB - We describe a patient presenting with a red eye who was found to have conjunctival non-Hodgkin's lymphoma of the mucosa-associated lymphoid tissue (MALT) type. PMID- 10474730 TI - Acute lymphoblastic leukaemia presenting with arthritis in an adult patient. AB - The earliest manifestations of leukaemia often include rheumatic signs and symptoms. Arthritis is a well recognised complication of leukaemia in children, but acute and chronic leukaemia may also cause arthritis in adults. Leukaemic arthritis may occur at any time during the course of leukaemia and may be the presenting manifestation. It should therefore be considered in the differential diagnosis of both childhood and adult rheumatic disease. We present an adult patient presenting with arthritis due to acute leukaemia. PMID- 10474731 TI - Calcium phosphate stones during long-term acetazolamide treatment for epilepsy. AB - We report a case of recurrent renal calculi containing calcium phosphate associated with long-term acetazolamide treatment for epilepsy. Unfortunately, the cause of stone formation was not recognised for many years, by which time irreversible renal damage had occurred. PMID- 10474732 TI - Quinine-mediated disseminated intravascular coagulation. PMID- 10474733 TI - A 60-year-old man with expressive dysphasia. PMID- 10474734 TI - A case of sympathetically maintained pain. PMID- 10474735 TI - Skin fragility and abnormal liver function tests. PMID- 10474736 TI - Syncope, the head ruling the heart. PMID- 10474737 TI - Tachycardia following low-tension electrocution. PMID- 10474738 TI - Collapse and hypothermia in an elderly woman in early summer. PMID- 10474739 TI - Compensated hypothyroidism. PMID- 10474740 TI - Compensated hypothyroidism. PMID- 10474741 TI - ACE inhibitors and insulin sensitivity. PMID- 10474742 TI - Inhibition of advanced glycated end-products. PMID- 10474743 TI - [Optical method of voltage measurement using voltage-sensitive dyes]. PMID- 10474744 TI - [Analysis of cerebrovascular circulation and neural transmission by SPECT]. PMID- 10474745 TI - [Basic knowledge of functional MRI applicable to daily practice]. PMID- 10474746 TI - The communicator and the professional. PMID- 10474747 TI - The importance of patient-physician communication: a patient's perspective. PMID- 10474748 TI - A catalyst for change in communication skills: the Canadian Breast Cancer Initiative. AB - A major focus of the Canadian Breast Cancer Initiative (CBCI) has been the development of the programming, partnerships and networks necessary to enhance communication skills within the physician-patient relationship. In this paper, the rationale for taking this approach and the scope of the activities that have been undertaken will be described, as well as the rationale for producing this series of papers. February has been chosen as the month of publication, as previous major activities of the CBCI in the area of physician-patient communication skills have been held in the month of February. It is hoped that February may become known as Communication Skills Month just as October is Breast Cancer Month: the gauntlet is thrown down to others to continue making February the month to reinforce the significance of physician-patient communication skills in the provision of quality health care. PMID- 10474749 TI - Evidence on patient-doctor communication. AB - This chapter covers important and well-studied aspects of patient-doctor communication. First the paper describes the lessons learned from studies about patients' satisfactions or dissatisfactions related to patient-doctor communication, making the point that complaints about doctors are usually due to communication problems and not technical competency issues. The next section of the chapter deals with time. It is often assumed that effective communication is inefficient. While this is not necessarily the case, the research results are complex and very interesting. The third part of the chapter covers communication in relation to patient adherence with the management plan recommended by the doctor. There is strong evidence that communication affects patient adherence and that there are four key aspects of communication that can enhance the patients' co-operation with the management plan. The final topic is patients' health. Twenty-two studies indicate the generally positive effect of key dimensions of communication on actual patient health outcomes such as pain, recovery from symptom, anxiety, functional status, and physiologic measures of blood pressure and blood glucose. PMID- 10474750 TI - Speaking of values: the ethics of communication. AB - The patient-doctor relationship is central to medicine. From this relationship, duties and obligations are derived; within this relationship, hopes and expectations are set. Communication is essential in this relationship and evidence-based efforts directed at improving it are a key element in improving care. At an even deeper level, critical reflection reveals fundamental values operating within and beneath this discourse that must be addressed if the goals of improving communication are to be achieved in a meaningful way. This is the stuff of the ethics of patient-physician communication. Because these values are so deeply embedded in the speech and actions of physicians, insights are best provided by those who are observers of the discourse rather than by participants. Observations from the history and sociology of physician communication, literary analysis and reflections on the illness experience by women provide important insights into the values and attitudes underlying physician communication, which must be taken into account in the education of physicians if the outcome is to benefit all participants. PMID- 10474751 TI - Medical education initiatives in communication skills. AB - Medical educators at undergraduate, postgraduate and continuing medical education levels acknowledge that communication is a fundamental medical skill. Responding to patient, professional and governmental advocates, as well as to advances in research on patient-physician communication and its teaching, some medical educators are in the process of starting new communication curricula, while others are working at expanding, integrating and further developing already well established programs. For most people working in this area, the question is no longer whether to teach and assess communication skills and attitudes but, rather, how to do so most efficiently and effectively. In order to enhance the development of communication curricula at all levels, we first provide a brief look at how communication education has become widely encouraged in many parts of the globe, and we set out the underlying assumptions that frame the teaching and learning of communication in medicine. We then summarize critical components common to many established communication curricula and identify a series of specific strategies for teaching communication skills. We include a chart that describes a sample of the wide variety of resources available to assist in the development and teaching of communication curricula in medicine. Finally, we consider gaps in current communication curricula and suggest the next steps and ideas for moving forwards. PMID- 10474752 TI - The role of medical organizations in supporting doctor-patient communication. AB - The clinical competence of physicians depends largely on the education, accreditation, certification and licensing programs offered by the various Canadian medical organizations. In virtually all of these, doctor-patient communication is a required element. Educational programs at all levels are subject to accreditation by a number of different organizations including undergraduate medical programs (Committee on Accreditation of Canadian Medical Schools), residency training (College of Family Physicians of Canada and Royal College of Physicians and Surgeons of Canada) and continuing medical education (CFPC and RCPSC). Doctor-patient communication is a key element in teaching at all levels. The two colleges also emphasize communications in the certification process. The provincial licensing authorities are aware of the importance of effective communication between physicians and patients. Several of the them have physician assessment programs, and recently they have started to assess a model of mandatory performance review. Both of these approaches assess physician patient communication. There is increasing pressure, with strong support from consumers, that some level of communication skills competency should be imposed by the licensing authorities. Most approaches to exposing physicians to communications focus on rewards rather than coercion but a number of possible schemes could be considered to promote communication skills. PMID- 10474753 TI - Canadian women's perspectives on ovarian cancer. AB - OBJECTIVE: To describe the perspectives of Canadian women living with ovarian cancer regarding their experiences with the disease. DESIGN: A cross-sectional survey of a convenience sample of Canadian women with ovarian cancer. SETTING: Survey questionnaires were sent to physicians in 26 cancer programs that treat women with ovarian cancer and to ovarian cancer self-help groups for subsequent distribution to women. PARTICIPANTS: Women diagnosed with ovarian cancer and able to read English or French. MAIN OUTCOME MEASURES: A variety of individual items in the survey related to information received, communication, physical and psychosocial symptoms, impact of illness and quality of life. RESULTS: A total of 315 women returned the survey. The average age of the respondents is 59 years. Each province and territory is represented in the sample. Over one-half of the women received a diagnosis of ovarian cancer within a month of seeking help for a concern and 85% had multiple treatment modalities. The majority of the women felt adequately informed (80%) and were satisfied with communication with their physicians (mean of 4.1 to 4.5 on a 5-point scale). A majority (62%) said that their lifestyle had changed as a result of their disease. Problems were experienced most frequently regarding side effects (58%), fear of recurrence (54%), sleeping difficulties (46%), bowel difficulties (44%), fear of dying (36%) and difficulty concentrating (32%). Many who experienced problems reported receiving inadequate help for them (16% to 49%). Quality of life was reported as significantly lower following the diagnosis and treatment of ovarian cancer (p = 0.0001). CONCLUSION: This study provides an important foundation for further investigation. There is a pressing need for research regarding the early identification of ovarian cancer and issues of support and coping. Care for women with ovarian cancer requires the expertise of a range of disciplines and community-based agencies working collaboratively as a team. PMID- 10474754 TI - Family physicians' perspectives on ovarian cancer. AB - OBJECTIVE: To describe the knowledge, practices and perspectives of Canadian family physicians regarding ovarian cancer. DESIGN: A mailed survey questionnaire was followed by a reminder card, a second mailing of the questionnaire and a final reminder card. SETTING: A national sample of family physicians was drawn randomly from the membership database of the College of Family Physicians of Canada. MAIN OUTCOME MEASURES: Knowledge related to ovarian cancer. Practices related to the screening and detection of ovarian cancer. Attitudes towards screening for ovarian cancer. Perceived role in the care of women at risk of, or diagnosed with, ovarian cancer. Perceived educational needs of physicians. RESULTS: A total of 1079 completed questionnaires were returned, providing a response rate of 56.6%. Although most family physicians were aware of the basic facts about ovarian cancer, there were knowledge limitations related to risk factors, familial ovarian cancer syndromes and symptoms. Practices related to asymptomatic women were found to be mostly in accord with current guidelines and recognized the prevailing lack of evidence for the effectiveness of tests. Areas that were troublesome included the role of screening in high-risk women and knowledge about available tests. Most family physicians indicated that they have an important role to play in the care of women after they have been diagnosed with ovarian cancer. They also expressed a high level of interest in obtaining additional information related to ovarian cancer. CONCLUSIONS: This study clearly shows that there is a need for additional research to assist with the development of evidence-based guidelines for women at increased risk of ovarian cancer and for women at no known risk. Pending more definitive evidence, interim guidelines could provide assistance to physicians currently having to make decisions in a context of massive uncertainty. Canadian family physicians would be interested in and would benefit from continuing medical education (CME) initiatives concerning ovarian cancer. PMID- 10474755 TI - Gynecologists' perspectives regarding ovarian cancer. AB - OBJECTIVE: To document the perspectives, practices and knowledge of Canadian gynecologists regarding ovarian cancer. DESIGN: A mailed survey questionnaire was followed by a reminder card and a second mailing of the questionnaire. SETTING: A sample of all gynecologists practising in Canada. MAIN OUTCOME MEASURES: Knowledge related to ovarian cancer. Practices related to the screening and detection of ovarian cancer. Attitudes towards ovarian cancer. Perceived role in the care of women at risk of, or diagnosed with, ovarian cancer. Perceived educational needs of gynecologists regarding ovarian cancer. RESULTS: A total of 504 completed questionnaires were returned, providing a response rate of 46%. Most gynecologists indicated that they knew the basic facts about ovarian cancer and risk factors. Practices related to asymptomatic, low-risk women were found to be mostly in accord with current guidelines and the lack of evidence for the effectiveness of tests. Practices regarding women with suspected early or late stage ovarian cancer varied, particularly with regard to referral to gynecologic oncologists. Many respondents indicated that they have an important role to play in the care of women after they have been diagnosed with ovarian cancer or referred to another specialist. Respondents also expressed interest in obtaining additional information about ovarian cancer. CONCLUSIONS: This study shows that there is a need for the development and dissemination of evidence-based guidelines regarding ovarian cancer. It also pinpoints areas where educational efforts could be directed. PMID- 10474756 TI - Complementary health practitioners' attitudes, practices and knowledge related to women's cancers. AB - OBJECTIVE: To document the attitudes, practices and knowledge of 3 groups of complementary practitioners (naturopathic doctors, chiropractors and massage therapists) regarding women's cancers in general and ovarian cancer specifically. DESIGN: A mailed survey questionnaire was followed by a reminder card and a second mailing of the questionnaire. SETTINGS: National samples were obtained for naturopathic doctors and chiropractors. The massage therapist sample was drawn from Ontario only because of the absence of a national listing of massage therapists. MAIN OUTCOME MEASURES: Practitioners reported response to patients' suspicious symptoms. Practitioners' perceptions of patients' motivations for seeking treatment. Practitioners' satisfaction with interactions with conventional practitioners. Practitioners' perceptions of their role in the care of women at risk of, or diagnosed with, cancer. Practitioners' perceptions of their knowledge regarding women's cancers. Practitioners' knowledge specific to ovarian cancer. RESULTS: A total of 894 completed questionnaires were returned, providing a response rate of 56%. The vast majority of practitioners who saw women with symptoms possibly related to cancer referred them to a family physician or a cancer specialist. Motivations that practitioners most frequently heard expressed by women seeking complementary treatments were "maximizing quality of life," "seeking natural approaches to healing" and "looking to stay well when disease is in remission." Most respondents were dissatisfied with patient-related communication with both family physicians and cancer specialists. The majority of complementary practitioners indicated that they have an important role to play in the postdiagnostic care of women with cancer. Considerable interest was expressed in further education concerning ovarian cancer. CONCLUSIONS: Whereas the professions reached through this survey differ in important ways from each other, they share an interest in being involved in the care of women with cancer, as well as an enthusiasm for the development of continuing professional education programs to help them better serve their clients. PMID- 10474758 TI - The creation of a database for cancer screening: is the consent of clients required? AB - Professional caregivers have an obligation to maintain the privacy and confidentiality of all personal information given to them by their patients or clients. Such information is to be released to a party who is not participating in the care of the patient only with the express consent of the patient. The question is whether or not the express consent of a client of an organized cancer screening program is ethically required before the client's personal information is entered into a centralized database. The arguments in favour of express consent are grounded largely in a desire to respect clients' privacy and autonomy fully. The arguments for the reliance on nonexpress, implied or tacit consent are based chiefly on the desire to provide clients with the full benefits of screening. It is suggested here that neither extreme position is satisfactory. A novel, balanced approach that would be sensitive to both electronic data storage and a preventive care situation is needed. Progress on this issue requires focused empirical research, a debate in both professional and public forums, and further critical analysis. PMID- 10474757 TI - The use of gemcitabine in non-small-cell lung cancer. Provincial Lung Cancer Disease Site Group. Provincial Systemic Treatment Disease Site Group. AB - GUIDELINE QUESTION: Is there a role for the use of gemcitabine in the treatment of patients with locally advanced or metastatic non-small-cell lung cancer (NSCLC)? OBJECTIVE: To make recommendations about the use of gemcitabine in the management of medically appropriate patients with stage IIIB-IV NSCLC. OUTCOMES: The outcomes of interest were survival, response rate, symptomatic response, response duration and toxicity. PERSPECTIVE (VALUES): Evidence was selected and reviewed by 2 members of the Provincial Lung Cancer Disease Site Group (DSG) of the Cancer Care Ontario Practice Guidelines Initiative. The practice guideline report was reviewed by the Provincial Lung Cancer DSG and by the Systemic Treatment Disease Site Group. These committees comprise medical and radiation oncologists, surgeons, pathologists, nurses, a psychologist, a medical sociologist and administrators. One community representative participated in the development of this practice guideline. QUALITY OF EVIDENCE: Five phase II studies of single-agent gemcitabine in advanced NSCLC were reviewed. Four of these are published as full reports. Two randomized phase II studies comparing single-agent gemcitabine with etoposide plus cisplatin were also reviewed. One of these studies is fully published. Seven phase II studies of gemcitabine in combination with cisplatin and I phase II study of gemcitabine in combination with ifosfamide were reviewed. Three randomized controlled trials (RCTs) and 1 randomized phase II study, published in abstract form, compared gemcitabine combination chemotherapy with cisplatin combination chemotherapy. An additional phase II study, published in abstract form, of gemcitabine as salvage therapy in previously treated patients was also included. BENEFITS: Four phase II studies of single-agent gemcitabine at a dose of 1000 mg/m2 or more showed a combined response rate of 19% (intention-to-treat analysis; 95% confidence interval [CI] 15% to 24%) or 21% (efficacy analysis; 95% CI 17% to 26%) in advanced NSCLC. Median survival ranged from 7 to 9 months. Improvement from baseline in cough, hemoptysis and dyspnea was comparable to what would be expected with radiation therapy and with standard combination chemotherapy regimens. Improvement from baseline in their performance status was reported in 52% of treated patients. The 2 randomized phase II studies reported equivalent response rates for gemcitabine compared with etoposide plus cisplatin; the response data were pooled, which resulted in a nonsignificant benefit for gemcitabine (common odds ratio [OR] 0.90; 95% CI 0.43 to 1.90; p = 0.78). Gemcitabine has most frequently been combined with cisplatin, yielding a combined response rate of 44% (intention-to treat; 95% CI 36% to 47%) or 45% (efficacy; 95% CI 39% to 51%) from 7 phase II studies. Median survival times ranged from 10 to 14 months. One phase II randomized study compared gemcitabine-cisplatin-vinorelbine vs. cisplatin epirubicin-vindesine plus lonidamine and demonstrated a higher response rate (62% vs. 35%) in favour of the gemcitabine combination. Three RCTs demonstrated increased response rates for the combination of gemcitabine-cisplatin over either cisplatin alone or other combination regimens [(gemcitabine-cisplatin 35% vs. etoposide-cisplatin 12%; p = 0.001), (gemcitabine-cisplatin 31% vs. cisplatin 9%; p = 0.0001), (gemcitabine-cisplatin 40% vs. mitomycin, ifosfamide, cisplatin 28%; p = 0.03)]. HARMS: The major dose-limiting toxicity is neutropenia. Despite this, infection rates are low. Significant adverse effects that have an impact on the patient's quality of life or require the discontinuance of treatment are reported to be less than with any other single agent or combination of agents. Grade 3 or 4 dyspnea has been reported to occur in fewer than 2% of cases and may be drug related. (ABSTRACT TRUNCATED) PMID- 10474759 TI - The Sociobehavioural Cancer Research Network: background and progress report. AB - The Sociobehavioural Cancer Research Network (SCRN) was established in 1994 by the National Cancer Institute of Canada (NCIC) with funding from the Canadian Cancer Society (CCS). The network was created to facilitate the development of behavioural science studies that would contribute to a fuller understanding of the cancer experience, from prevention through detection, treatment and post treatment (including palliative care). This article describes the nature of network research, the development and organization of the Sociobehavioural Cancer Research Network and the challenges it faces. PMID- 10474760 TI - Tumour epidermal growth factor receptor, erbB-2 and cathepsin D in node-negative invasive breast cancer: their impact on the selection of patients for systemic adjuvant therapy. AB - OBJECTIVE: To determine the feasibility and the economic impact of tumour EGFR, erbB-2 and cathepsin-D measurements in women with node-negative breast cancer. DESIGN: Consecutive tumour samples received at a regional steroid receptor laboratory from patients with node-negative breast cancer were evaluated with commercially available kits to determine EGFR, erbB-2 and cathepsin-D levels. SETTING: All node-negative patients whose tumours were submitted to the steroid receptor laboratory from November 1992 to March 1994 were included (n = 142). A control group of concurrent node-negative breast cancer patients from the London Regional Cancer Centre (LRCC) database were also evaluated to determine the representativeness of our sample. MAIN OUTCOME MEASURE: To determine the proportion of patients who were positive for the 3 newer prognostic factors relative to their risk of relapse. RESULTS: We found 75 positive values in 69 patients (48.6%). We demonstrated that each factor identified a different high risk subgroup. Epidermal growth factor receptor (EGFR) positivity (> 10 fmol/mg protein) was found in 16.3% of patients, with 19.9% of patients positive for erbB 2 (> 250 units/mg protein) and 17.3% positive for cathepsin D (> 70 pmol/mg protein). Between 10% and 23.2% more node-negative patients currently seen in a regional cancer centre could be offered systemic adjuvant chemotherapy based on a single positive new factor. CONCLUSIONS: These tumour evaluations are straightforward using material already available in a regional steroid receptor laboratory or on tumour tissue available to pathologists. The economic impact is minimal; the 1995 cost of performing all 3 evaluations is Can$425-616 (US$304 440) per patient treated depending on the number of assays per run. Prospective clinical trials incorporating tumour EGFR, erbB-2 and cathepsin D are feasible and economically viable. PMID- 10474761 TI - Communication between family physicians and oncologists: qualitative results of an exploratory study. AB - OBJECTIVE: To study communication between family physicians (FPs) and oncologists, and to look at the factors that may influence FP involvement in cancer care. DESIGN: This survey design uses a qualitative methodology, where the data are analyzed using a modified grounded theory approach. SETTING: This was a multisite study using 14 focus groups of FPs, followed by structured telephone interviews with 116 FPs in 6 different Canadian provinces. MAIN OUTCOME MEASURE: Interview questions were used to explore the actual and desired roles of FPs in cancer care, and the quality of communication with oncologists with reference to a particular cancer patient in the FPs' practice. RESULTS: Physicians providing cancer care must consider complex psychosocial and biomedical factors, more so than with other chronic diseases, and so written communication alone is inadequate. Family physicians require face-to-face and/or telephone communication with the oncologist to negotiate their respective roles, and to discuss the patient's prognosis and the effectiveness of proposed treatments. Family physicians expressed a desire to become more involved in all stages of cancer care in both the biomedical and psychosocial aspects, and to help better define their roles throughout the illness trajectory. CONCLUSION: These results suggest opportunities to improve the communication, coordination and comprehensiveness of shared cancer care provided by family physicians and oncologists in different clinical settings. PMID- 10474763 TI - Staging in cancer. PMID- 10474764 TI - The importance of alternative therapies to the public. PMID- 10474762 TI - Use of dexrazoxane as a cardioprotectant in patients receiving doxorubicin or epirubicin chemotherapy for the treatment of cancer. The Provincial Systemic Treatment Disease Site Group. AB - GUIDELINE QUESTIONS: 1) Should dexrazoxane be used routinely in patients with advanced or metastatic cancer who are at risk of developing cardio toxicity when receiving chemotherapy containing doxorubicin or epirubicin? 2) Do the available data support the use of dexrazoxane when anthracyclines are being used in the adjuvant setting for patients at risk of developing cardiotoxicity? OBJECTIVE: To make recommendations regarding the use of dexrazoxane to prevent cardiotoxicity in patients with nonhematological malignancies who are receiving anthracycline- containing chemotherapy. OUTCOMES: Clinical and subclinical cardiotoxicity, noncardiac toxicity and impact on efficacy outcomes such as response and overall survival are considered. PERSPECTIVE (VALUES): Evidence was selected, reviewed and synthesized by 2 members of Cancer Care Ontario's Systemic Treatment Disease Site Group (STDSG), formerly the Systemic Treatment Program Committee. Drafts of this document have been circulated and reviewed by members of the STDSG. The STDSG comprises medical oncologists, pharmacists, supportive care personnel and administrators. Community representatives did not participate in the development of this guideline, but they will be included in future guidelines. QUALITY OF EVIDENCE: Seven randomized controlled trials (RCTs), 2 with placebo control, were available for analysis. BENEFITS: Data for clinical cardiotoxicity from 6 trials were pooled (n = 1070). The meta-analysis indicated that the risk of experiencing clinical cardiotoxicity was significantly reduced by dexrazoxane (risk ratio 0.24; 95% confidence interval [CI] 0.11 to 0.52; p = 0.00031). There was no significant benefit shown in individual trials for objective response or survival. HARMS: One of the RCTs revealed a significantly lower objective response rate in the dexrazoxane arm. However, a meta-analysis of objective response across 5 trials of breast cancer patients (n = 818) did not confirm this effect (odds ratio 0.85; 95% CI 0.61 to 1.18; p = 0.33). The use of dexrazoxane increased the incidence of myelosuppression and other noncardiac toxicities, but these were generally mild. PRACTICE GUIDELINE: The evidence supports the use of dexrazoxane to provide protection against the cardiotoxicity associated with conventional-dose doxorubicin in patients with advanced but anthracycline sensitive cancer, in whom the continued use of anthracycline-containing chemotherapy is indicated in the opinion of the treating physician and who have received 300 mg/m2 or more of doxorubicin. The evidence supports the use dexrazoxane to provide protection against the cardiotoxicity associated with conventional-dose epirubicin in patients with advanced but anthracycline sensitive cancer, in whom the continued use of anthracycline-containing chemotherapy is indicated in the opinion of the treating physicians. There are no data indicating the optimal cumulative dose of epirubicin at which dexrazoxane should be instituted. For doxorubicin, use of dexrazoxane is recommended after the cumulative dose reaches 300 mg/m2 (i.e., 55% of the recommended maximum). A similar formula could be used for epirubicin, that is, institution of dexrazoxane when the cumulative dose of epirubicin reaches 550 mg/m2, as the recommended maximum cumulative dose in Canada is 1000 mg/m2. Preclinical studies did not show any cardioprotectant effect for dexrazoxane when used with mitoxantrone, and no clinical studies have been done. Therefore, dexrazoxane is not recommended for use with mitoxantrone. There is no evidence for or against the use of dexrazoxane in the adjuvant setting for any tumour type. Because of concerns that dexrazoxane may reduce the efficacy of anthracyclines, and because data are not yet available on long-term toxicities, further studies should be performed before the drug is used in this setting. PMID- 10474765 TI - Cancer patients' expectations of the role of family physicians in communication about complementary therapies. AB - OBJECTIVE: To explore cancer patients' experiences with and expectations of the role of family physicians in communication about complementary therapies. DESIGN: Focus group interviews. SETTING: British Columbia Cancer Agency and the University of Calgary. PARTICIPANTS: Sample of 14 cancer patients using complementary therapies comprising 3 focus groups (2 in BC and 1 in Alberta). RESULTS: The role of family physicians in discussing complementary therapies varied from being extraneous to patients' decision making to being a partner in making decisions about cancer treatments. Patients expected their physicians to be supportive, caring, kind and to show an interest in them. They also expected their physicians to be accepting and nonjudgmental regarding complementary therapy use and to reinforce a sense of hope. Most patients did not expect their doctors to have extensive knowledge about these therapies. All patients strongly felt the need to seek information themselves. In addition, they felt that believing in the type of therapies they were using was very important. CONCLUSION: Family physicians can play an important role in exploring and discussing complementary therapies with their patients. It is important that they are made aware of their patients' needs in this area. PMID- 10474766 TI - Evaluation of the "Sunny Days, Healthy Ways" sun safety CD-ROM program for children in grades 4 and 5. AB - Computer-based sun safety instruction has many advantages that may be attractive to health educators in schools. An educational multimedia computer program on sun safety was produced on CD-ROM for children in grades 4 and 5, which was based on the "Sunny Days, Healthy Ways" sun safety curriculum (SDHW). Its effects on children's sun safety knowledge, attitudes and behaviour were evaluated with 162 students in 8 fourth and fifth grade classes in a randomized pretest-posttest 2 x 2 factorial design. Children interacting with the CD-ROM program showed significant improvements in knowledge (p = 0.007). The effect on knowledge may have indirectly improved children's sun protection (r = 0.201, p = 0.013), even though the CD-ROM program did not directly increase sun protection (p > .05) or improve attitudes (p > .05). The CD-ROM program may be a cost-effective and administratively acceptable sun safety instructional strategy, however, like many short prevention strategies, it will be most successful at conveying information on sun safety to children. PMID- 10474767 TI - Compliance with radiation treatment guidelines in a provincial setting. AB - Since 1979, consensus guidelines have been produced for radiation treatment practice in British Columbia. These guidelines have been revised, updated and circulated to all radiation oncologists periodically. A comprehensive computer database for all patients receiving radiation treatment in the province was established in 1984. Between 1985 and 1996 inclusive, 7667 prostate cancer patients and 9748 breast cancer patients received primary radical radiation treatment or adjuvant postsurgical treatment. Palliative treatments and the treatment of other disease sites are not included in this review. Compliance of these treatments with the published guidelines is reported. Over 98% of patients completed treatment as planned and, in the last 3 years, more than 90% of patients received a guideline dose to either the prostate or the breast. Compliance was less in the treatment of the regional lymph nodes in breast cancer. Practice in prostate cancer tended to precede the changes in the guidelines. This was not the case for breast cancer. PMID- 10474768 TI - Have diagnostic practices contributed to trends in leukemia incidence and mortality among Canadians? AB - OBJECTIVE: To outline the patterns and temporal trends in leukemia, regarding incidence and mortality, in Canada since 1970. DESIGN: A descriptive analysis of trends in incidence and mortality by age, sex, time period and leukemia subtype, using change-point modelling and log-linear regression. SETTING: Data from all provincial and territorial cancer registries. MAIN OUTCOME MEASURE: Incidence and mortality rate per population of 100,000. RESULTS: The overall age-adjusted incidence rates for all males and females increased from 12.3 and 7.3 per 100,000 in 1970-1972 to 14.6 and 9.0 in 1979-1981, then decreased to 13.2 and 8.3 in 1991 1993, respectively. The overall age-adjusted mortality rates decreased from 9.1 and 5.7 per population of 100,000 in 1970-1972 to 8.3 and 4.8 in 1993-1995 for males and females, respectively. The incidence of acute leukemias decreased between 1970 and 1993. A sharp increase in the incidence of chronic leukemias from 1978 to 1980 was observed in the older age group. Mortality rates also showed a small increase from 1979 to 1989 in seniors. CONCLUSION: The increase in the incidence of chronic leukemias among older subjects was probably due to improvements in diagnosis and changes in registration practices, whereas the decrease in the incidence of acute leukemias was probably a real change attributable to environmental factors. Further investigation is needed to clarify whether and to what extent diagnostic practices contributed to the increased detection of chronic leukemias among elderly Canadians. PMID- 10474770 TI - Development of peripheral autonomic synapses: neurotransmitter receptors, neuroeffector associations and neural influences. AB - 1. The functional innervation of autonomic target tissues occurs early during development, at a time when both the nerves and post-synaptic target tissues are still differentiating. 2. Physiological responses appear soon after the arrival of the first fibres when uptake and release mechanisms within the nerves are already functional. Initial responses differ from those in the mature animal, both in the form and, frequently, in the subtypes of receptors involved. 3. Results of a number of studies suggest that the initial expression of neurotransmitter receptors during development is largely independent of neural influences. Changes recorded in neurotransmitter receptor expression during development appear to be similarly independent of neural influences. 4. While signal transduction pathways coupling adrenergic neurotransmitter receptors to effector responses appear to develop independently of the nerves, the efficient coupling of muscarinic receptors often requires the action of the neurotransmitter, acetylcholine. 5. During the period of synapse formation, the neural plexus continues to expand. While developing varicosities can release the neurotransmitter, the capacity for neurotransmitter retention appears to be restricted. Developmental changes in the neurotransmitters that produce functional responses, while well known in the sweat glands, may also be seen in more subtle forms in other target tissues. 6. Ultrastructural studies suggest that close physical associations between the membranes of the release sites of the developing nerves and the target cells may form early during development when physiological responses are still immature. These close associations could enable more specific reciprocal interactions between nerves and target cells involving known and novel growth factors, neuropeptides and cytokines important in shaping the mature synaptic characteristics. PMID- 10474769 TI - An epidemiological review of red cell transfusions in cancer chemotherapy. AB - OBJECTIVE: The objective of this chart review was to determine the frequency of transfusion and prevalence of anemia (hemoglobin result < 100 g/L) in patients receiving chemotherapy. DESIGN: This study was a retrospective review of medical charts. SETTING: Patients receiving chemotherapy were included from 12 tertiary care comprehensive cancer centres across Canada. MAIN OUTCOME MEASURE: The primary study outcome measure was red blood cell transfusion rate, controlling for patient variables. RESULTS: The 616 patients included had started chemotherapy in January-June 1992. For each subject, data collection finished 4 weeks after the end of the first regimen or after a maximum follow-up period of 26 weeks. Seventy-two patients (12%; 95% confidence interval 9.5% to 14.5%) were transfused for anemia (reasons other than blood loss), and 28% (95% confidence interval 24.5% to 31.5%) of the subjects were anemic during treatment. The univariate analyses of transfusion for anemia yielded significant associations with prognostic factors. In the multivariate analyses, platinum (odds ratio [OR] = 6.69) and anthracycline (OR = 3.56) chemotherapy, baseline hemoglobin (OR = 0.96) and disease stage (OR = 1.72) were statistically significant contributors. CONCLUSION: In this patient cohort, red blood cell transfusion was infrequent (12%). However, patient groups at high risk of transfusion could be identified, with platinum-based chemotherapy being the most significant contributing factor. The information obtained from this multicentre study may prove helpful in developing supportive care guidelines for the management of chemotherapy-related anemia requiring transfusion. PMID- 10474771 TI - Evolution of haemoglobin function: molecular adaptations to environment. AB - 1. Nearly 1000 mutations have been described for human haemoglobin (Hb), many of which result in subtle changes to the oxygen transport system. Similar changes have occurred over the course of animal evolution resulting in a diversity of functional attributes which appear to correlate the availability of oxygen in the environment with metabolic demand. 2. At an early stage in evolution, worm-like animals had large, polymeric aggregations of Hb subunits circulating through primitive circulatory systems and some possessed monomeric Hb in blood cells functioning as an oxygen store. 3. The circulating vertebrate red blood cell provides an environment allowing haem units to interact among themselves and with various organic phosphates to allow a responsive and highly regulated system of gas transport. During metazoan evolution the burden of physiological regulation has shifted from the cells to organ systems, as endothermy and aerial breathing permit a relatively constant environment. 4. An understanding of the adaptive possibilities of Hb has helped us to understand the ontogeny of oxygen transport and to interpret recently described functional properties of human embryonic haemoglobins. PMID- 10474772 TI - Human erythrocyte but not brain acetylcholinesterase hydrolyses heroin to morphine. AB - 1. In human blood, heroin is rapidly hydrolysed by sequential deacylation of two ester bonds to yield first 6-monoacetylmorphine (6-MAM), then morphine. 2. Serum butyrylcholinesterase (BuChE) hydrolyses heroin to 6-MAM with a catalytic efficiency of 4.5/min per mumol/L, but does not proceed to produce morphine. 3. In vitro, human erythrocyte acetylcholinesterase (AChE) hydrolyses heroin to 6 MAM, with a catalytic efficiency of 0.5/min per mumol/L under first-order kinetics. Moreover, erythrocyte AChE, but not BuChE is capable of further hydrolysing 6-MAM to morphine, albeit at a considerably slower rate. 4. Both hydrolysis steps by erythrocyte AChE were totally blocked by the selective AChE inhibitor BW284c51 but were not blocked by the BuChE-specific inhibitor, iso-OMPA (tetraisopropylpyrophosphoramide). 5. The brain synaptic form of AChE, which differs from the erythrocyte enzyme in its C-terminus, was incapable of hydrolysing heroin. 6. Heroin suppressed substrate hydrolysis by antibody immobilized erythrocyte but not by brain AChE. 7. These findings reveal a new metabolic role for erythrocyte AChE, the biological function of which is as yet unexplained, and demonstrate distinct biochemical properties for the two AChE variants, which were previously considered catalytically indistinguishable. PMID- 10474773 TI - Hyperlipidaemia and the progression of nephropathy in OLETF rats: effect of angiotensin-converting enzyme inhibitor, enalapril. AB - 1. The influence of angiotensin-converting enzyme (ACE) inhibitor is investigated in enalapril on renal function in Otsuka Long-Evans Tokushima Fatty (OLETF) rats, an animal model of spontaneously non-insulin-dependent diabetes (NIDDM). 2. Enalapril (5 mg/kg) or vehicle was administered once daily by gastric gavage to 22-week-old male OLETF rats for 32 weeks. Blood pressure, albuminuria, creatinine clearance, plasma glucose, serum insulin and lipids were determined before and during the treatment. Renal haemodynamics was examined at the end of the treatment. 3. Enalapril lowered blood pressure mildly but significantly. In the vehicle-treated rats, urinary albumin excretion increased from 0.75 +/- 0.16 mg/mg creatinine (Cr) to 8.65 +/- 0.78 mg/mg Cr. Enalapril significantly blunted the development of albuminuria from 0.66 +/- 0.12 mg/mg Cr to 5.19 +/- 0.67 mg/mg Cr (P < 0.008) without significant influence on creatinine clearances. Enalapril also significantly blunted the rise in serum cholesterol and triglyceride prior to the development of massive albuminuria. Enalapril did not affect bodyweight, plasma glucose or insulin levels. Renal haemodynamics assessed by inulin and p aminohippuric acid clearances were similar in both groups at the end of the treatment. 4. These results reconfirmed that the ACE inhibitor has protective effects on nephropathy in NIDDM. Massive albuminuria was preceded by increase in serum lipids in OLETF rats, which supports the view that hyperlipidaemia exacerbates glomerular injury in chronic renal disease. Enalapril attenuated the rise in serum lipids, suggesting that the beneficial effects of the compound on renal injury in OLETF rats might also be mediated through the action of affecting serum lipids. PMID- 10474774 TI - Renal nerve and alpha 2-adrenergic action during acute volume expansion in the anaesthetized rat. AB - 1. The effects of renal denervation and alpha 2-adrenoceptor blockade on diuresis and natriuresis in response to acute volume expansion (VE) were investigated in anaesthetized rats. 2. In normal rats, a 30 min intravascular VE of up to 7.5% of bodyweight caused a significant diuresis and natriuresis. 3. Denervation enhanced the diuretic effect of VE by 39%, while alpha 2 blockade by 12 mumol/kg per h rauwolscine significantly inhibited the response by 61% in innervated kidneys and 43% in denervated kidneys. 4. Similar results were found for natriuresis in both innervated and denervated kidneys. 5. The results demonstrate that renal sympathetic nerve activity inhibits part of the increase in sodium and water excretion caused by VE, since denervation increased excretion under these conditions. The contribution of renal nerves to the control of renal function, however, decreases during the response to VE. 6. alpha 2-Adrenoceptor action enhanced the diuretic and natriuretic effects of VE by mechanisms otherwise unrelated to renal nerve activity. PMID- 10474775 TI - Sex- and age-related changes in urinary contents of alpha 1-microglobulin and ulinastatin in mice. AB - 1. Using two murine strains, ICR/Jcl and senescence-accelerated-prone inbred mouse 1 (SAMP1), age-associated changes in urine indices (e.g. urine volume, creatinine contents, contents of alpha 1-microglobulin (alpha 1M) and ulinastatin (UT) and the relation of urinary contents of alpha 1M and UT were investigated. 2. Sex-related differences in the indices were not observed in ICR nor in SAMP1 mice. 3. Urine indices per 24 h of ICR mice at 6 and 14 months of age were higher than those at 3 months of age, although indices of SAMP1 mice did not change with ageing. Urinary contents of alpha 1M and UT in ICR mice at 6 and 14 months of age were higher than those in SAMP1 mice. However, contents of alpha 1M and UT expressed as the contents per creatinine did not differ between these two strains. 4. In the relation between urinary contents of alpha 1M and UT, a positive correlation was displayed both in ICR and SAMP1 mice, and the regression slope did not significantly differ with ageing in these two strains. 5. These results suggest that ageing per se is not a factor which affects the relation of urinary contents of alpha 1M and UT. PMID- 10474776 TI - Physical and biochemical predictors of death in an Australian aboriginal cohort. AB - 1. Predictors of natural and cardiovascular death were examined in a cohort from a remote Australian Aboriginal community with high mortality rates. The cohort was marked by high prevalences of diabetes (17%), hypertension (19% diastolic blood pressure (BP) > 90 mmHg), obesity (16% body mass index (BMI) > 30), albuminuria (26% albumin/creatinine ratio (ACR) > or = 34 g/mol) and renal failure (26% calculated glomerular filtration rate < 80 mL/min). 2. Relationships between baseline characteristics and subsequent mortality were examined using multivariate techniques. 3. Albuminuria, diabetes and hypertension (but not smoking dyslipidaemia or obesity) were all markers for increased risk of natural and especially of cardiovascular death. Age- and sex-adjusted relative risk [95% CI] of natural deaths were 4.3 [1.9-9.5] for overt albuminuria (ACR > or = 34, 3.2 (range 1.6-6.5) for diastolic BP > or = 100 and 3.7 (range 1.5-8.9) for diabetes. 4. The relative risk associated with albuminuria was independent of diabetes and hypertension. Albuminuria was more common than either diabetes or hypertension and represents a target for intervention to reduce not only progression to renal disease, but also overall mortality. PMID- 10474777 TI - Low dose indapamide plus perindopril combination effects on cardiovascular structure and function in genetic hypertension. AB - 1. Although the fixed combination preparations of thiazide and angiotensin converting enzyme inhibitor are gaining wide acceptance in clinical practice, data on the basic pharmacology of the combinations are relatively limited. The long-term structural and functional effects of a fixed low dose (0.24 + 0.76 mg/kg per day) combination of indapamide + perindopril (I + P,S5590) is spontaneously hypertensive rats (SHR) were examined in the present study. 2. Male SHR (10-12 weeks) were treated with I + P or vehicle for 8 weeks. The blood pressure and heart rate were monitored by weekly measurements. At the end of the treatment period, left ventricular, aortic and intramyocardial coronary arteriole structures were assessed. Contractile and relaxant properties of mesenteric arteries were determined by wire-myography. 3. Indapamide + perindopril combination caused a significant lowering of both systolic (P < 0.001) and diastolic (P < 0.001) blood pressures. Left ventricle plus septum:bodyweight ratio (P < 0.001), aortic medial cross-sectional area (P < 0.05) and media:lumen ratios (P < 0.005) were all significantly reduced by I + P treatment. In contrast, the effect of I + P on intramyocardial coronary vascular structure did not reach statistical significance. There was some improvement in endothelium independent vasorelaxation of mesenteric vessels but contractile responses to noradrenaline and calcium were unaffected by treatment. 4. In summary, a low dose I + P combination treatment of SHR partly normalizes both systolic and diastolic blood pressures. Cardiac and larger vessel hypertrophy was reversed but intramyocardial coronary arteriole structure was not as readily regressed by the end of the study. PMID- 10474778 TI - Adrenocorticotrophic hormone-induced hypertension in the rat: effects of the endothelin antagonist bosentan. AB - 1. The effects of the endothelin antagonist bosentan on adrenocorticotrophic hormone (ACTH)-induced hypertension were examined in the conscious male Sprague Dawley rat. 2. In order to confirm endothelin antagonism, 18 rats were randomly divided into two groups: receiving either (i) endothelin-1 (0.125, 0.25, 0.5 and 1 nmol/kg, i.v.); or (ii) endothelin-1 at these doses following bosentan (100 mg/kg gavage) and mean arterial pressure recorded (study A). Subsequently, 40 male rats (320 +/- 5 g) were randomly divided into four groups (n = 10): (i) Sham (0.9% saline, s.c.) + 5% acacia gum gavage; (ii) ACTH (500 micrograms/kg per day, s.c.) + 5% acacia gum gavage; (iii) Sham injection + bosentan (100 mg/kg per day) gavage; or (iv) ACTH + bosentan. Six control days (C1-C6) were followed by 11 treatment days (T0-T10). Systolic blood pressure, water intake, urine volume, food intake and bodyweight were measured every second day (study B). 3. Bosentan significantly attenuated the endothelin-1-induced blood pressure rise at 0.125 nmol/kg (P < 0.05), but not at higher doses. 4. Bosentan at a dose which attenuated endothelin-1-induced blood pressure increase had no effect on either blood pressure or metabolic parameters in ACTH-treated rats. 5. These results suggest that endothelin does not play a major role in ACTH-induced hypertension. PMID- 10474779 TI - Changes of urinary dopamine excretion early after balloon mitral commissurotomy in mitral stenosis. AB - 1. In order to investigate the changes of reduced urinary free dopamine excretion (uDA) in heart failure, 15 patients with symptomatic mitral stenosis were investigated on their uDA, endogenous creatinine (Cr) clearance, urinary excretion of sodium (UNaV), fractional excretion of sodium (FENa), plasma noradrenaline (pNA) and plasma L-dopa concentration before and early after percutaneous transvenous mitral commissurotomy (PTMC) by the clearance study. The delivery of L-dopa to renal proximal tubules (plasma L-dopa x Cr clearance), and the conversion ratio of plasma L-dopa to urinary dopamine in the kidney [uDA/(plasma L-dopa x Cr clearance)] were also estimated. 2. After successful PTMC, uDA, UNaV and FENa showed a significant but incomplete improvement and the changes of uDA were correlated positively with those of cardiac index (CI) (r = 0.665, P < 0.01), not with changes of pulmonary wedge pressure. While plasma L dopa and plasma L-dopa x Cr clearance improved, uDA/(plasma L-dopa x Cr clearance) was not significantly changed early after PTMC. 3. From these results, it was suggested that reduced uDA tended to increase incompletely in relation with functional recovery of heart, and that increased plasma L-dopa and a delivery of L-dopa to renal proximal tubules have some positive role on urinary dopamine excretion, at least, early after PTMC. PMID- 10474780 TI - Muscle venous PO2 and VO2 are linearly related in repetitive tetanic contractions of canine muscle during hypoxic hypoxia. AB - 1. It has previously been shown that perfusion with high O2-affinity-erythrocytes decreases venous PO2 (PVO2) and decreases O2 uptake (VO2) in contracting muscle at the same O2 delivery (arterial O2 concentration x flow). A linear VO2-PVO2 relationship has been obtained with a VO2-axis intercept, suggesting that, during this type of hypoxia, VO2 is composed of a PVO2-dependent and -independent VO2. However, the VO2-PVO2 relation during hypoxic hypoxia has not been examined. 2. To clarify this relation, PVO2 and VO2 have been measured in contracting gastrocnemius (1 Hz trains of 0.2 s isometric tetani) under both normoxic and hypoxic conditions during 5 min of stimulation. 3. Venous O2 changes proportionally with O2 delivery. Each VO2-PVO2 relation was linear, with the mean described by the equation VO2 = 5.06 + 0.41 x PVO2 (n = 6, r = 0.81, P < 0.05). The VO2-axis intercept was significantly different from zero (P < 0.05). 4. These results were similar to those obtained during hypoxia induced by high O2-affinity erythrocytes. We conclude that there is a linear relationship between PVO2 and VO2 above the VO2-axis intercept, regardless of the type of hypoxia. PMID- 10474781 TI - Renal haemodynamic and excretory responses to bradykinin in anaesthetized dogs. AB - 1. Effects of bradykinin (BK) on renal haemodynamics and urine formation were examined in anaesthetized dogs. 2. Renal arterial infusion of BK at doses of 5 or 50 ng/kg per min produced dose-dependent increases in renal blood flow (RBF), without affecting systemic arterial pressure or glomerular filtration rate. There were also significant and dose-dependent increases in urine flow (UF), urinary excretion of sodium (UNaV) and fractional excretion of sodium (FENa) and decreases in urine osmolality during BK infusion. 3. Renal haemodynamic and excretory responses to the BK infusion were completely abolished by the simultaneous administration of Hoe 140 (icatibant, 100 ng/kg per min intrarenally), a selective BK B2-receptor antagonist. 4. In the presence of NG nitro-L-arginine (NOARG; 40 micrograms/kg per min intrarenally), a nitric oxide (NO) synthase inhibitor, BK-induced renal vasodilative and natriuretic effects were markedly attenuated, although responses of UF and urine osmolality to BK remained unchanged. The water diuretic effect of BK was abolished in dogs given both NOARG and ibuprofen (12.5 mg/kg bolus injection plus 12.5 mg/kg per h of sustained infusion intravenously), a cyclooxygenase inhibitor. 5. These results clearly indicate that renal haemodynamic and excretory responses to BK were mediated exclusively by the B2-receptor. Renal vasodilative and natriuretic responses are mainly linked to NO generation, while both NO and prostaglandin biosynthesis are involved in the BK-induced water diuresis. PMID- 10474782 TI - Expression and regulation of the sodium-calcium exchanger in cardiac microvascular endothelial cells. AB - 1. The sodium-calcium exchanger (NCX) plays an important role in Ca2+ homeostasis. In the heart, NCX participates in the control of contraction and relaxation and in large vessel endothelial cells some data suggest that NCX could influence nitric oxide (NO) generation. In this context, the cardiac microvasculature has received considerable attention as a mediator of myocardial performance, via the release of paracrine acting factors such as NO. Therefore, the aim of the current study was to characterize NCX expression and regulation in cardiac microvascular endothelial cells (CMEC). The NCX expression was also examined in neonatal ventricular cardiomyocytes where aspects of its function and regulation have been well characterized. 2. The presence of functional NCX in CMEC was confirmed by the presence of a consistent rise in intracellular Ca2+ concentration ([Ca2+]i) in response to removal of extracellular Na+. Furthermore, NCX mRNA expression was readily detectable in CMEC. 3. In order to examine the role of possible physiological regulators of NCX expression, the effect of intracellular Ca2+ loading, caused by 24 h exposure to 10 mumol/L ouabain, was investigated. In Ca(2+)-loaded CMEC, there was a substantially greater rise in [Ca2+]i during exposure to Na(+)-free buffer: 33 +/- 6 versus 124 +/- 25 nmol/L% (P < 0.05), consistent with increased protein expression. Consistent with these findings, northern blot analysis confirmed the presence of a two-fold increase in NCX mRNA in these cells. 4. These data indicate the presence of functional NCX in CMEC and identify [Ca2+]i as a potential physiological regulator of expression. PMID- 10474783 TI - Reduced dietary fat intake increases parasympathetic activity in healthy premenopausal women. AB - 1. Hypercholesterolaemia has been associated with decreased heart rate variability, a measure of cardiac parasympathetic activity. However, the effect of perturbation of the lipid profile on autonomic function has not been examined systematically. 2. The effects of short-term dietary lipid modification on autonomic function are studied in 25 normotensive, non-smoking, premenopausal women with normal bodyweight. Subjects consumed either a low (L, 25%) or high fat (H, 40%) diet for 2 weeks in an open, randomized, cross-over manner with a 2 week washout. 3. Baroreflex sensitivity was determined by gating beat-to-beat heart period (RR) interval and continuous non-invasive blood pressure recordings. Heart rate variability measures of cardiac parasympathetic nervous system activity were obtained in the time (standard deviation of RR intervals, root mean square of successive differences (rMSSD)) and frequency (high frequency power) domains. All assessments were made at the same timepoint in the menstrual cycle. 4. Both low density lipoprotein-cholesterol and high-density lipoprotein-cholesterol decreased significantly (P < 0.05) with increased dietary fat intake (H, 2.7 +/- 0.1 vs L, 2.2 +/- 0.1; H, 1.3 +/- 0.1 vs L, 1.1 +/- 0.1 mmol/L, respectively) as did mean arterial pressure (H, 78.1 +/- 1.5 vs L, 74.3 +/- 1.5 mmHg). Weight was unchanged by dietary lipid intake (H, 62.6 +/- 8.5 vs L, 62.3 +/- 8.3 kg, P = NS). 5. There was a significant increase in rMSSD (H, 29.6 +/- 3.4 vs L, 38.8 +/- 6.4 msec, P < 0.05) and natural logarithm of high frequency power following low fat diet (H, 4.4 +/- 0.2 vs L, 4.8 +/- 0.3 msec2, P = 0.01). Baroreflex sensitivity also increased following the low fat diet (H, 13.91 +/- 2.2 vs L, 16.9 +/- 3.2 msec/mmHg, P = 0.23). 6. Short-term dietary lipid modification can significantly increase cardiac parasympathetic nervous system activity in healthy premenopausal women. These changes in autonomic status appear to be independent of changes in bodyweight and may be of clinical relevance considering the prognostic implications of heart rate variability in cardiovascular disease. PMID- 10474784 TI - The role of digitalis in the treatment of heart failure. AB - Over the past 200 years, digoxin has been commonly used to treat patients with congestive heart failure. Clinical trials have demonstrated the benefits of the use of digoxin on exercise tolerance, ejection fraction, and neurohormone production. The Digoxin Investigators Group trial has recently provided strong evidence for the long-term benefits of digoxin on morbidity for patients with heart failure. This article will review the evidence of the benefits of digoxin and its current role in the treatment of patients with congestive heart failure. PMID- 10474785 TI - Angiotensin-converting enzyme inhibitors: congestive heart failure and beyond. AB - Angiotensin-converting enzyme (ACE) inhibitors were introduced in the early 1980s as a novel class of agents to treat hypertension. Since that time, they have been proven to be a powerful tool in the treatment of symptomatic left ventricular systolic dysfunction. In this article we will summarize the results from clinical trials using ACE inhibitors in the treatment of heart failure and after myocardial infarction. We will discuss some of the mechanisms postulated to account for the beneficial effects associated with ACE inhibitor treatment. Finally, we will highlight some recent controversies in the use of ACE inhibitors. PMID- 10474786 TI - Beta-adrenergic blocking agents: past, present, and future perspectives. AB - The 'proof of concept' of beta-blockade for heart failure (i.e. that the pharmacologic actions of beta-blockers are beneficial) is now firmly established, as the treatment of heart failure has progressed from using positive inotropic stimulation, via drugs with no direct effect on cardiac function, to beta blockers with negative intropic effects. This review addresses some remaining issues regarding beta-blockade in heart failure. The mechanism of action of beta blockers in heart failure is more likely to be improved intrinsic cardiac myocyte function and prevention or reversal of remodeling, than restoration of beta adrenergic signal transduction. The role of the differentiating characteristics of beta-blockers is not clear at this time, and there is no compelling evidence to select one agent over another on the basis of individual drug properties. Recent reports suggest that beta-blockers reduce the combined risk of all-cause mortality and hospitalizations by about 30-35%. These results are heavily influenced by experience with carvedilol, but other agents tested include metoprolol, bucindolol, bisoprolol, and nebivolol. Responsiveness to beta blockers is not related to patients' age, sex, or race, or to the etiology or severity of heart failure. Beta-blockers are currently recommended as adjunctive treatment in patients who remain mildly to moderately symptomatic while receiving added digitalis, diuretics, and angiotensin-converting enzyme inhibitors. Existing gaps in our knowledge must be filled in order to achieve optimal clinical application of beta-blockers. Ongoing studies will provide much of the information required. The role of beta-blockers will probably expand as we improve our understanding of the pathophysiology of heart failure, and especially of the remodeling process. PMID- 10474787 TI - Angiotensin II receptor blockers. PMID- 10474788 TI - Growth hormone as a potential future therapy for congestive heart failure. PMID- 10474789 TI - Potential future therapies: natriuretic peptides. AB - Congestive heart failure has become a major public health problem. A hallmark of this syndrome is neurohumoral activation, with elevation of natriuretic peptides, in particular atrial natriuretic peptide and brain natriuretic peptide of myocardial origin, which occurs with the onset of ventricular dysfunction. The natriuretic peptide system is important in cardiorenal regulation, specifically in the integrated control of intravascular volume and arterial pressure to maintain optimal circulatory integrity. Several therapeutic approaches have been established to mimic or potentiate the unique cardiovascular and renal beneficial actions of these peptides during heart failure. Recent investigations have also established a diagnostic utility for the natriuretic peptides to diagnose early asymptomatic left ventricular dysfunction. Thus, diagnostic and therapeutic use of the natriuretic peptides is emerging as a new strategy to delay the natural history of progressive heart failure from its earliest phase to chronic congestive heart failure. PMID- 10474791 TI - Potential future therapies for heart failure: gene transfer of beta-adrenergic signaling components. AB - Congestive heart failure (CHF), despite the improved prevention and treatment modalities adopted for cardiovascular disease over the past two decades, remains a significant therapeutic challenge. Efficacious therapies are few, and death rates from CHF continue to rise. Recent advances in our understanding of the molecular basis of CHF have given rise to experimental animal models demonstrating related genetic phenotypes, which further elucidate cellular mechanisms involved in the pathogenesis of the failing heart. Studies involving transgenic mice have elucidated novel potential gene therapy interventions aimed at the genetic modification of beta-adrenergic signaling in the heart. This review will briefly discuss beta-adrenergic signaling in CHF, while focusing on potential gene therapy strategies to improve the performance of the failing heart. PMID- 10474790 TI - Thyroid hormone and thyroid hormone analogues in the treatment of heart failure. AB - The thyroid hormone analogue DITPA is a promising potential new treatment for heart failure. Although the mechanism of action is incompletely determined, it is clear that DITPA improves systolic as well as diastolic function. It is also clear that the effects of DITPA are intrinsic to the muscle and not the result of changes in the structure or geometry of the left ventricle. On the basis of these experimental studies, we applied to the USA Food and Drug Administration for an Investigational New Drug application to study the use of DITPA in patients. These studies are currently in progress. While we await the outcome of these clinical trials, it is important to emphasize that even if the end-point is not a new drug to treat heart failure, our investigations are based on a systematic evaluation integrating biochemistry and physiology. We believe that this is the way to approach the problem of developmental pharmacology. PMID- 10474792 TI - Carotid intima-media thickness of young coronary patients. AB - BACKGROUND: Although a greater than normal intima-media thickness (IMT) has been found in older coronary patients, the data for younger patients are lacking. OBJECTIVE: To determine the carotid IMT in patients with premature myocardial infarction. METHODS: We measured IMT (common and internal carotid, carotid bifurcation) in 30 coronary patients, aged 30-50 years (mean 46 years), who had survived myocardial infarctions 1-9 years (mean 6 years) earlier, and in 30 age matched men without clinically evident coronary heart disease (controls) by B mode ultrasonography. Blood levels of lipoproteins, glucose, iron and transferrin, fibrinogen, tissue plasminogen activator (t-PA) antigen level and activity and plasminogen activator inhibitor (PAI-1) antigen level and activity were also determined. RESULTS: IMT in all segments of carotid arteries in the patients was significantly greater than that in the controls (P < 0.0001). The presence of atherosclerotic plaques was correlated to greater than normal carotid IMT. Other risk factors displaying statistically significant correlations to mean carotid IMT were t-PA antigen level and activity, PAI-1 antigen level and level of high-density lipoprotein cholesterol. CONCLUSIONS: The significant association between carotid IMT and coronary heart disease suggests that carotid and coronary atherosclerosis evolve simultaneously. Hence carotid IMT can be used as a predictor of coronary atherosclerosis. PMID- 10474794 TI - One-year clinical follow-up with the serpentine balloon expandable stent: report of the first 100 patients. AB - BACKGROUND: We recently reported a high technical and 30-day clinical success rate among the first 100 patients treated with the tubular, serpentine design, stainless steel, balloon-expandable stent (beStent) in Israel. The present study examined the clinical results in these patients after the first year. METHODS: Seventy-eight men and 22 women were included in the study. Previous myocardial infarction, bypass surgery and percutaneous transluminal coronary angioplasty had occurred in 52%, 12% and 26% of the patients, respectively. Diabetes mellitus was present in 30 patients and hypertension in 34 patients. One hundred and forty eight stents of 15, 25, and 35 mm lengths were used. The indications for stenting were suboptimal results (n = 85), bailout conditions (n = 10) or for the prevention of restenosis (n = 8), and lesion types were A (n = 10), B1 (n = 29), B2 (n = 20), and C (n = 44). All patients were clinically monitored with regular visits at 1, 3, 6, 9 and 12 months. RESULTS: Overall, the 12-month event-free survival rate was 82%. Subacute thrombosis occurred in two patients. There were two non-cardiac deaths, one O-wave myocardial infarction, six elective bypass surgeries and 12 target lesion revascularizations. Event-free survival was significantly higher for those with lesions shorter than 15 mm than for those with lesions longer than 15 mm (90% versus 67%, P = 0.003), and for women compared with men (96% versus 78%, P = 0.02). CONCLUSIONS: The initial experience with the beStent shows favorable long-term results with an overall event rate of 18% for this subset of relatively complex lesions; higher event rates were observed for longer lesions. PMID- 10474793 TI - Impairment of the blood-brain barrier: a potential surrogate delineating the determinants of cerebral bleeding caused by fibrinolytic drugs. AB - BACKGROUND: Intracranial bleeding is encountered in some patients with acute myocardial infarction treated with fibrinolytic drugs, and especially in patients with occult cerebral vasculopathy. In order to determine whether pharmacologically induced plasminemia is a determinant, and whether impairment of the blood-brain barrier can serve as a marker of risk, we studied spontaneously hypertensive stroke-prone rats (SHRSP) genetically disposed to cerebral vasculopathy. METHODS: In order to simulate the induction of plasminemia in patients treated with fibrinolytic drugs for acute myocardial infarction, three intravenous injections of human plasminogen and human tissue-type plasminogen activator (t-PA) were administered to the rats at 2-h intervals (12 mg plasminogen plus 60 micrograms t-PA, 6 mg plus 30 micrograms t-PA, and 0.5 mg plus 2.5 micrograms t-PA), and serial blood samples were assayed for plasmin. One hour after the final intravenous injection, the brain was perfusion-fixed with 4% formaldehyde, and the blood-brain barrier integrity was assessed by localization of immunoglobulin G (IgG) using fluorescein-conjugated goat anti-rat IgG and confocal microscopy. RESULTS: Transient plasminemia followed each injection of plasminogen and t-PA. Intracranial extravasation of IgG was observed in nine of 11 SHRSP treated with t-PA and plasminogen. None of seven SHRSP injected with vehicle alone exhibited extravasation (chi 2 = 6.37, P = 0.01). CONCLUSIONS: The results indicate that the blood-brain barrier in SHRSP is susceptible to impairment secondary to pharmacologically induced plasminemia and is predisposed to bleeding. The preparation developed may facilitate the delineation of relative risk of injury to cerebral vessels with thrombolytic therapy. PMID- 10474796 TI - Bibliography. Current world literature. PMID- 10474795 TI - A cellular paradigm for the failure to increase vascular endothelial growth factor in chronically hypoxic states. AB - BACKGROUND: Angiogenesis, or new blood vessel formation, is the physiological adaptation of a tissue to hypoxia or ischemia. However, this compensatory response to hypoxic stress in vivo is often insufficient. In many of the conditions in which the angiogenic response to tissue hypoxia is insufficient, such as chronic critical limb ischemia or myocardial hibernation, the hypoxic stress is chronic and persistent, lasting for days or even months. Vascular endothelial growth factor (VEGF) has been demonstrated in vivo and in vitro to be the principal mediator of hypoxia-induced angiogenesis. We propose that the lack of compensatory angiogenesis in response to tissue hypoxia in many clinical syndromes characterized by chronic hypoxia is due to a failure to induce VEGF appropriately. METHODS AND RESULTS: Heart or liver cells were grown under conditions of chronic hypoxia, returned to a normoxic environment, and then rechallenged with hypoxia. We found that the hypoxic induction of VEGF mRNA was markedly blunted using this algorithm. Furthermore, transient transfection studies using the VEGF promoter containing an oxygen-responsive enhancer element failed to show induction in cells pretreated by subjection to chronic hypoxia. CONCLUSIONS: Hypoxic pretreatment results in a blunting of the ability of a cell to induce VEGF in response to subsequent episodes of hypoxia. This may provide a rationale for the inadequate amount of compensatory angiogenesis seen in many chronic ischemic disorders. PMID- 10474797 TI - [Low-threshold potential-controlled calcium channels--their properties and functions]. AB - Low voltage-activated calcium channels discovered 15 year ago are unique molecular structures which play a specific role in the functioning of different types of excitable cells. This conclusion is supported by recent identification of special channel-forming 1- subunits which after expression in artificial cellular systems generate transmembrane currents characteristic for low-voltage activated calcium channels. At the same time, analysis of recent experimental data indicate the existence of several subtypes of low voltage-activated channels which differ in the characteristics of their permeability, kinetics and pharmacological sensitivity. The expression of different subtypes of low-voltage activated channels is found to be highly coordinated, on one hand, with different phases of the ontogenetic development of excitable cells and, on the other, with their fulfillment of specific functions. Therefore the search for selective modulators of the activity of these channels is an extremely important goal for modern investigations. PMID- 10474798 TI - [Neurohumoral control of protein biosynthesis during aging]. AB - It has been shown in the experiments on adult (6-8 mo) and old (26-28 mo) Wistar rats that in aging, due to an electrical stimulation of the hypothalamus, the liver induction of thyroxine aminotransferase, tryptophan-pyrrolase, glucose-6 phosphatase and fructose-1, 6-diphosphatase decreases. In old rats, the biosynthesis of RNA fractions gets activated at later periods. In contrast to adult animals, the electrical hypothalamic stimulation did not induce any marked changes in chromatin fractions ratio and transcriptional processes in old rats. With aging, it may happen that the target cells cannot respond to adequate stimulations, while the central (hypothalamic, in particular) mechanisms cannot realize them. In aging, the influence of the hormones (insulin, testosterone, thyroxin and hydrocortisone) on the synthesis of proteins-invertors, regulating plasmatic membrane state Is weakened. Following surgical denervation of the liver in old rats, the less marked changes in RNA and protein synthesis and lesser influences on monooxygenase induction were found in old rats. All these observations indicate an impairment of the neural control over protein biosynthesis in senescence. Also, the axonal transport of proteins is delayed in old rats that may influence the nervous cell aging. Owing to all these shifts, the plastic provision of body integrative reactions is impaired. PMID- 10474799 TI - [Volume-sensitive anion conductivity in a human prostatic carcinoma cell line]. AB - For the first time membrane anion current activated in response to the cell swelling following exposure to hypotonic solution has been found in human prostate carcinoma cell line LNCaP. The development of hypotonicity-evoked current required 5 to 10 min to the maximum and was almost completely reversible upon return to the solution with normal osmolarity. Not absolute selectivity for Cl- ions of the respective cell volume-sensitive channels has been shown. PMID- 10474800 TI - [The action of pilocarpine on the accommodation properties of isolated phasic neurons of the rat hippocampus]. AB - Using of voltage and current clamp methods in conditions of perforated membrane fragment, we investigated the accommodation properties of isolated neurones from different zones of hippocampus. The investigated neurones could be separated into two main groups concerning their responses to prolonged pulses of suprathreshold depolarization--neurones which after generation of several action potentials (AP) stopped their generation (phasic, F-neurones), and (tonic, T-neurones), which did not accommodate and continued AP generation during entire depolarizing pulse. Comparison of ionic current composition in neurones of both groups has shown, that T-neurones demonstrate more expressed inactivation of outward currents that can be due to the presence of a fast inactivating A-type potassium current, or decreased inactivation of inward currents. Pilocarpine, well-known epileptogenic compound increased accommodation and induced significant slow-down of outward current inactivation in F- neurones. It can be suggested that the presence of A type potassium current and/or a slowly inactivating component of inward current can play main role in determining the accommodation effect in hippocampal neurones. PMID- 10474801 TI - [Variations in the ionic currents as dependent on the size of the sensory neurons in newborn rats]. AB - In the somatic membrane of acutely isolated newborn rat dorsal root ganglion neurons four inward and three outward currents were separated on the basis of their different voltage dependence and kinetic properties. The capacity of their membrane was measured for 136 investigated neurons. Neurons were separated according to capacities of their somata for three subpopulations: small 26.5 pF, medium 41.5 pF and large 61.4 pF. Neurons with TTX-insensitive sodium current were from medium and, partially, from small subpopulations. Neurons with low threshold calcium current were from large and, partially, from medium subpopulations. PMID- 10474802 TI - [Changes in calcium signaling in mammalian nociceptive neurons in diabetes mellitus]. AB - The changes in neuronal Ca2+ homeostasis were studied on dorsal horn neurons from spinal cord rat slices and freshly isolated dorsal root ganglion neurons of mice in control condition and under streptozotocin (STZ)-induced diabetes. The cytoplasmic free Ca2+ concentration ([Ca2+]i) was measured using fura-2 and indo 1 based microfluorimetry. The recovery of depolarization-induced [Ca2+]i increase was delayed in diabetic neurons compared with normal animals. The amplitude of calcium release from caffeine-sensitive endoplasmic reticulum calcium stores became significantly smaller in diabetic neurons. The participation of mitochondria in [Ca2+]i homeostasis was determined by investigation of changes which occurred after addition of mitochondrial protonophore (CCCP) to the extracellular solution. In control cells 10 (M CCCP applied before membrane depolarization induced an increase of the amplitude of depolarization-induced [Ca2+]i transients and disappearance of their delayed recovery, indicating the participation of mitochondria in fast uptake of Ca2+ ions from the cytosol during the peak of the transient and subsequent slow release them back during its decay. In neurons from diabetic animals the increase of the peak transient amplitude under the action of CCCP became diminished, and the delayed elevation of [Ca2+]i disappeared in small size neurons. We conclude that streptozotocin-induced diabetes is associated with prominent changes in the mechanisms responsible for [Ca2+]i regulation in neurones of the nociceptive system, which presumably include a slow down of Ca2+ elimination from the cytoplasm by endoplasmic reticulum and mitochondria. PMID- 10474803 TI - [The effect of SR 142801, a blocker of tachykinin NK-3 receptors, on synaptic transmission in the rat caudal mesenteric ganglion]. AB - Effects of tachykinin NK-3 receptors antagonist SR 142801 on the synaptic transmission in the rat's caudal mesenteric ganglion were investigated using extracellular recording of compound action potentials (CAPs) from the ganglion nerves. It is shown that antagonist of NK-3 receptors SR 142801 (2.5.10(-7) mol/l) decreased the amplitude and square of CAPs obtained by stimulating of intermesenteric nerve (IMN) and leading off from the colonic nerve, after high frequency stimulation of IMN. It is suggests that tachykinin NK-3 receptors take part in SP-ergic synaptic transmission through the rat's caudal mesenteric ganglion. PMID- 10474804 TI - [The pharmacological blockade of the cardiac and gastric enteric ganglia by a new parasympatholytic]. AB - The blocking effects of a newly synthetized compound N-decyltropine bromide (IEM 1556), on pentagastrin-, carbachol- and histamine-induced gastric secretion in chronic dogs, on stress-induced changes in gastric mucosa in rats, on vagus induced effect in heart and on arterial blood pressure in rats were studied. The effects were compared with those produced by hexamethonium, a conventional ganglionic blocking agent. IEM-1556 inhibited gastric secretion and acid output for a much longer time than hexamethonium did. It also effectively protected gastric mucosa against stress-induced erosions and hemorrhages. IEM-1556 much more effectively blocked the vagus-induced reduction of the heart rate than hexamethonium did, and, in contrast to latter, did not reduce the arterial blood pressure. The results suggest that IEM-1556 is a highly selective blocking agent for parasympathetic and enteric ganglia versus sympathetic ganglia. PMID- 10474805 TI - [The effect of melatonin on photoperiod changes in the glutathione system of the brain under acute hypoxia]. AB - The effect of acute hypobaric hypoxia against on the background of single-shot melatonin administration on the activity of glutathione ferments system (glutathione peroxidase, glutathione S-transferase and glucose-6-phosphate dehydrogenase) and the contents of reduced glutathione and malondialdehyde in the supernatant of juvenile male of white rats forebrain was investigated under three conditions of lighting--natural conditions of lighting in the spring-summer period of year, constant illumination and constant darkness during the one week. The constant illumination enhanced the lipid peroxidation with simultaneous decreasing in the activity of glutathione peroxidase in control animals. The constant illumination enhanced the lipid peroxidation with simultaneous decrease in the activity of glutathione peroxidase in control animals. The constant darkness reduced the intensity of free-radical oxidation improving the antioxidant neuron protection. The melatonin decreased the contents of malondialdehyde, raised the activity of glutathione enzyme system and eliminated the negative influence of constant illumination on forebrain antioxidant system in intact animals. The acute hypoxia resulted in the increase of lipid peroxidation and reduced the activity of antioxidant enzymes, which were most expressed in constant light conditions. The melatonin administration before of 30 minutes of acute hypoxia modeling reduced the intensity of oxidizing stress, which was generated by acute hypoxia. PMID- 10474806 TI - [An electrophysiological evaluation of the role of the olfactory analyzer in brain integrative activity]. AB - Summary electrical activity of olfactory bulb and amygdala of unrested rats with reversible anosmia during conditioning with application of light and odor as conditioned stimuli was studied. It was shown, that recovery of capacity for reception of odor wasn't connected with appearance in electrical activity of investigated structures of high-frequency high-amplitude components. Additional investigations of neuronal isolation of olfactory bulb and discovery of correlative changes of power of high-frequency components in dependency on level of common excitation give the possibility to consider this rhythm as one of the mechanisms of nonsensory modulator influences of olfactory analyser. PMID- 10474807 TI - [The demonstration of steady-state Ca2+ influx into the cells of the salivary glands in Chironomus plumosus L. larvae and its role in basal secretion]. AB - Ca2+ content in Chironomus plumosus salivary gland tissue we determined using Ca(2+)-sensitive dye arsenazo III and protein concentration in medium--by Lowry method. It was showed correlation between increasing of Ca2+ content in gland and basal secretion. So, basal secretion is Ca(2+)-dependent process. Owing to adding to medium verapamil, diltiazem and nifedipine (10(-4) M) took place decreasing of Ca2+ content and secretion. Since decreasing of Ca2+ entry in cells owing to blocators action did't exceed 53% (in the case of nifedipine) we assumed that permanent Ca2+ entry formed by potential- and receptor-operated channels. It was established dependence of continuous Ca2+ entry in cells, basal secretion and effectiveness of blockade of Ca(2+)-channels by nifedipine on the [Ca2+]e. Therefore, through the exocrine secretory cells membrane of Chironomus plumosus larvae salivary gland carry out continuous diffuse Ca2+ entry for maintain of basal secretion by cells. This Ca2+ entry provide by population of open Ca(2+) channels, sensitive to blocators of potential dependent of Ca(2+)-conduction plasmatic membrane. PMID- 10474808 TI - [The suppression of c-fos protein expression in the islands of Calleja in damage to the mesostriatal dopaminergic system of the brain]. AB - We found activation c-fos protein in the NO synthase-containing neurons within the islands of Calleja (96.46 +/- 16.03 fos-immunoreactive nuclei per 50 mm section) in control and suppression of this activation after unilateral 6 hydroxydopamine lesions of dopaminergic mesostriatal system of brain (14.31 +/- 4.59 nuclei per section). Since the neuronal activity in the islands of Calleja is associated with the blood supply changes in the basal ganglia we propose that the suppression of c-fos protein expression in this structure after lesions of dopaminergic mesostriatal system reflects the disorders its blood supply. PMID- 10474809 TI - [Pathological changes in thrombocyte-vascular and coagulative hemostasis under the influence of lead chloride and their correction by using a synthetic analog of prostacyclin]. AB - It has been established that plumbum chloride possesses a marked toxic action on the membranes of the formed elements, in particular, the thrombocytes, activates the release of the factors of thrombocyte aggregations, resulting in an increase of their adhesive and aggregation properties. The research studies the influence of the stable prostacycline analogue MM-706 and thrombocytic-vascular, as well as coagulative hemostasis in rats at a background of plumbum intoxication. We have discovered a marked protective action of MM-706 on the thrombocytes which is manifested through a decrease of the index of a spontaneous thrombocyte aggregation and percentage of adhesive thrombocytes, normalization of the general potential of hemocoagulation and time characteristics of a thromboelastogram. PMID- 10474810 TI - [The effect of nitric oxide donors on the respiratory rhythm of an artificially superfused preparation of the brain of newborn rats]. AB - The influence of an exogenous nitric oxide (NO) donor, sodium nitroprusside, and endogenous NO biosynthesis substrate, L-arginine, on integrated respiratory rhythm recorded from the phrenic nerve of the semi-isolated medullo-spinal preparations of the 3-to-4-days old rats was studied. Applications of 10.0 mcM of these drugs led to significant increase in the duration and amplitude of the inspiratory discharges (ID) in the phrenic nerve and to decrease of ID frequency. After the rostral ventrolateral (RVL) medulla (projectionally corresponds to "M" chemosensitive zone) had been separated by transection, such applications began to evoke a rise of ID frequency without significant changes of the duration and amplitude. Thus, we found the involvement of NO in the tonic inhibition of the respiratory rhythm by RVL medullary structures in the neonatal rats. PMID- 10474812 TI - [Endothelial cells during cultivation (a comparative analysis of methodological approaches)]. AB - In the paper various methods for isolation and cultivation of endothelial cells are compared on the basis of the own data. The comparative analysis of the methods for isolation of endothelial cells from the vessel explant and by enzymatic treatment of aorta or coronary vessels is carried out. The methodical approaches to obtaining pure culture of endotheliocytes are being analysed. PMID- 10474811 TI - [The effect of medikhronal on the biogenic amine level in the brain structures and blood of animals with alcohol dependence]. AB - Peculiarities of changes in the content of biogenic amines in the brain structures (hypothalamus, midbrain and new cortex) and blood of rats under alcohol intoxication and formation of alcohol dependence were investigated, and the effect of medichronal and its components on the mentioned processes was found out. It has been established that accumulation of acetaldehide plays a considerable role in the mechanism of formation of alcohol dependence. Changes in the level of ethanol (E) and AcA in blood after the arrival of exogenic E to the organism evokes phase dynamics of content of biogenic amines in the blood and brain sections. Medichronal components, binding and inactivating AcA, as well as taking the mutually supplementing effect on synthesis of biogenic amines, promote normalization of the mentioned processes and elimination of alcohol dependence. PMID- 10474813 TI - [A method for performing a study and assessment of the individual neurodynamic properties of human higher nervous activity]. AB - The proposed methods are directed at unification of conducting and assessing neurodynamic properties of the higher nervous activity of a human that are related to the processing of visual information of various complexity levels. It should be considered that the conducting of examinations in maximum close conditions of the same tests and assessment criteria will increase the possibilities and the value of the analysis of various experimental materials. PMID- 10474815 TI - Mutant manifestation: the time factor in somatic mutagenesis. PMID- 10474814 TI - [Interneuronal signaling mediated by the trans-synaptic diffusion of neurotransmitters]. AB - A great number of modern neurophysiological studies give evidence about the possibility of the extrasynaptic diffusion of the released neurotransmitters. Such observations challenge the established view on synapses as isolated communication channels between pre- and postsynaptic neurones. In the present review article, we consider the latest experimental data on the interneuronal cross-talk mediated by extrasynaptic diffusion of L-glutamate (L-Glu) and gamma amino-butyric acid (GABA). Possible physiological significance of this phenomenon is also discussed. PMID- 10474816 TI - Evaluation of the mouse lymphoma tk assay (microwell method) as an alternative to the in vitro chromosomal aberration test. AB - In order to evaluate the utility of the mouse lymphoma assay (MLA) for detecting in vitro clastogens and spindle poisons and to compare it with the in vitro chromosomal aberration test (CA), we conducted an international collaborative study of the MLA that included 45 Japanese laboratories and seven overseas laboratories under the cooperation of the Ministry of Health and Welfare of Japan and the Japanese Pharmaceutical Manufacturer's Association. We examined 40 chemicals; 33 were reportedly positive in the CA but negative in the bacterial reverse mutation assay, six were negative in both assays and one was positive in both. We assayed mutations of the thymidine kinase (TK) locus (tk) of L5178Y tk +/- mouse lymphoma cells using the microwell method. According to our standard protocol, cells were exposed to the chemical for 3 h, cultured for 2 days and TK deficient mutants were expressed in 96-well plates under trifluorothymidine. Each chemical was coded and tested by two or three laboratories. Among the 34 CA positive chemicals, positive MLA results were obtained for 20 and negative results were obtained for nine. The remaining five chemicals were inconclusive or equivocal because of discrepant inter-laboratory results or reproduced discrepant results, respectively. Among the six CA-negative chemicals, one was negative in the MLA, two were positive and three were inconclusive. Thus, the MLA could detect only 59% (20/34) of CA-positive chemicals. We concluded that the MLA was not as sensitive as the CA. Some MLA-negative chemicals evoked positive responses in the CA only after long continuous treatment. These might also be genotoxic in the MLA with long continuous treatment. Improvement of the MLA protocol, including alteration of the duration of the treatment, might render the MLA as sensitive as the CA. PMID- 10474817 TI - The need for long-term treatment in the mouse lymphoma assay. AB - The L5178Y tk +/- mouse lymphoma assay (MLA) has been widely used as a genotoxicity test for the detection of mutagens and clastogens. The standard MLA, as well as other mammalian cell gene mutation assays, usually employs a short treatment period (3-6 h). Our previous report, however, suggested that such short treatments may be insufficient for detecting some clastogens and spindle poisons. For the present study, we introduced and evaluated a longer treatment (24 h) in the MLA. We examined 15 chemicals which were evaluated as negative or inconclusive in the short-term study. Cells were exposed to the chemical for 24 h without S9 mix, cultured for 2 days and then thymidine kinase-deficient mutants were selected in 96-well microtiter plates under trifluorothymidine. Eleven chemicals yielded positive responses in the 24 h treatment MLA. They included nucleoside analogs (2'-deoxycoformycin and dideoxycytidine), a base analog (1,3 dimethylxanthine) and spindle poisons (colchicine and vinblastine sulfate), all of which do not directly affect DNA, but bring about mutations and chromosome alterations through nucleoside metabolism and chromosome segregation. Because the mutagenicities of these non-DNA targeting chemicals appear to be cell cycle dependent, treatment extending over more than one cell cycle may be required for their effect. Combining results from the present and previous studies, 31 of 34 (91%) chromosome aberration-positive chemicals exhibited positive responses in the MLA, suggesting that the sensitivity of the MLA with 24 h treatment periods approaches that of the chromosome aberration test. PMID- 10474818 TI - gamma-Glutamyl transferase (GGT) deficiency in the GGTenu1 mouse results from a single point mutation that leads to a stop codon in the first coding exon of GGT mRNA. AB - GGTenul, a recently described genetic murine model of gamma-glutamyl transferase (GGT) deficiency, was induced by the point mutagen N-ethyl-N-nitrosourea and is inherited as an autosomal recessive trait. The phenotype of systemic GGT deficiency suggested a mutation site within the cDNA coding region which is common in all GGT transcripts. To identify this site, total lung and kidney RNA was isolated from normal and mutant mice, amplified by RT-PCR using GGT-specific primers, cloned as two overlapping approximately 1 kb GGT cDNA fragments, sequenced and compared with that in the literature. A single base pair substitution was identified in the coding region at position 237, where thymidine became adenine, and this mutation replaced a leucine codon, TTG, with a termination codon, TAG. This mutation site was confirmed in mutant genomic DNA by PCR using primers that flanked the predicted site and spanned the intron between the common GGT non-coding exon and the first GGT coding exon. This PCR product was sequenced directly with the secondary 3' PCR primer, the mutation site identified and the protocol then utilized to genotype animals. In addition to this mutation, the steady-state level of GGT mRNA in mutant kidney is reduced 3 fold compared with the control. Heterodimeric GGT protein is not detectable by western blot in either whole kidney homogenate or a microsomal membrane fraction. The steady-state mRNA level of gamma-glutatmyl cysteinyl synthetase was unchanged in mutant mice compared with normal, but that of heme oxygenase-1 and Cu,Zn-SOD was induced 4- and 3-fold, respectively. Hence, the GGTenul mouse model of GGT deficiency results from a single point mutation in the first coding exon of GGT mRNA and the resulting impairment in glutathione turnover induces oxidative stress in the kidney. PMID- 10474819 TI - Inverse restriction site mutation (iRSM) analysis. Mutation detection involving the formation of restriction enzyme sites in target genes. AB - This paper describes a rapid screening procedure for the detection of DNA sequence changes resulting in the creation of new restriction enzyme sites. The basic methodology involves the identification of the conversion of one restriction site into another by mutagenesis. The selective removal of the wild type sequences by digestion with a restriction enzyme acting on the wild-type sequence increases the sensitivity beyond that of PCR-RFLP analysis (10(-4)-10( 5) detectable here). In this paper we describe the rapid detection of induced in vivo mutations transforming the ApaI restriction site present in intron 6 of the mouse p53 gene to a unique AvaII site. The potential application of this method in other genes and organisms as a rapid screen for induced mutations is discussed. PMID- 10474820 TI - Effect of cytochalasin B on the induction of chromosome missegregation by colchicine at low concentrations in human lymphocytes. AB - The aim of the present work was to investigate the possible interference of cytochalasin B (cyt B) with low concentration treatment with colchicine in the induction of chromosome/chromatid loss and micronuclei in human lymphocytes mitotically activated in vitro. Thus, cells from a single female donor were treated with colchicine (10 or 25 nM, from 24 h after PHA addition to fixation at 66 h) either in the presence or absence of cyt B. Single lagging chromosomes/chromatids were scored in bipolar ana-telophases and greater damage (disrupted and c-anaphases) was scored in cells at anaphase. Micronuclei were scored in the first 4000 nuclei observed in both cyt B-treated (in mononucleate and binucleate cells) and untreated cultures. With the same criterion, FISH analysis was performed on 2000 nuclei where chromosome 7 and 11 centromeric DNA probes were used in pairs. Our results showed that: (i) the frequency of laggards and of micronuclei increased with colchicine concentration but in the presence of cyt B there was a lower frequency of both (with a mean reduction of approximately 49%); (ii) FISH analysis showed a colchicine concentration-dependent increase in nuclei with three spots for chromosome 7; (iii) a colchicine concentration dependent increase in tetraploid cells was observed. This increase was particularly remarkable (5-fold) in cells grown in the presence of cyt B compared with cyt B-untreated cells. The observed 'cyt B effects' can be explained if it is assumed that in cytokinesis-blocked cells there is a shorter distance between the poles. As a consequence: (i) laggards would be engulfed in the nearest daughter nucleus with a consequent lower induction of micronuclei; (ii) segregating sister chromatids in heavily impaired anaphases would not travel a sufficient distance to give rise to two daughter nuclei, leading to an increased frequency of polyploid nuclei. PMID- 10474821 TI - Noscapine hydrochloride disrupts the mitotic spindle in mammalian cells and induces aneuploidy as well as polyploidy in cultured human lymphocytes. AB - Noscapine hydrochloride is a centrally acting antitussive opium derivative widely used in cough suppressants. Recent studies have reported that noscapine is a potent inducer of polyploidy but not of aneuploidy in vitro. To obtain more comprehensive information about the cytogenetic effects of this compound, we treated cultured human lymphocytes (HPL) and Chinese hamster ovary (CHO) cells with various concentrations of noscapine hydrochloride. Using a differential staining technique noscapine was shown to disrupt the mitotic spindle at concentrations < 5 micrograms/ml in both cell types. The use of multicolor fluorescence in situ hybridization (FISH) on noscapine-treated human lymphocytes showed a dose-dependent induction of hyperdiploidy of chromosome 1 but not of chromosomal breakage in the 1cen-q12 region under in vitro conditions, indicating that noscapine is specifically inducing numerical chromosomal aberrations. FISH with probes targeting different chromosomes revealed that noscapine is capable of inducing both polyploidy and true hyperdiploidy. Our results show that noscapine, by disrupting the function of the mitotic spindle, has the ability to induce aneuploidy and not uniquely polyploidy as previously reported. By using these types of molecular cytogenetic techniques, it should be possible to evaluate the ability of noscapine to induce aneuploidy in the upper intestinal tract in vivo. PMID- 10474822 TI - Dual-colour FISH analysis to characterize a marker chromosome in cytochrome P450 2B1 recombinant V79 Chinese hamster cells. AB - Recombinant V79 Chinese hamster cell lines have been constructed for the expression of useful functions, e.g. cytochromes P450, in order to study metabolism-dependent toxicity. Recombinant cell lines are derived as single clones upon gene transfer and selection from the parental V79 cell line. It is of fundamental importance for a reliable application of the recombinant cell lines to show that the original biological characteristics of the parental line are maintained. As part of these efforts, fluorescence in situ hybridization with Chinese hamster chromosome-specific DNA libraries was performed in order to identify the origin of the chromosomes from which a marker chromosome present in the recombinant cell line V79MZr2B1 was derived, which could not be identified by standard cytogenetic techniques. PMID- 10474823 TI - Comparison of the aneugenic effect of vinorelbine and vincristine in cultured human lymphocytes. AB - Vinca alkaloids are used clinically against a variety of hematological and solid tumors. These compounds interact with tubulin subunits to prevent microtubule assembly, inducing abnormal chromosome segregation in dividing cells and causing aneuploidy. The vinca alkaloid vincristine sulfate (VCR) and the semisynthetic analog vinorelbine (VRB) were studied by analysis of micronuclei (MN) in cultured human lymphocytes using the cytokinesis block method. Furthermore, fluorescence in situ hybridization with a human alphoid satellite pancentromeric DNA probe was used to detect centromeres in isolated MN of VRB- or VCR-treated lymphocytes. At all the doses tested, both chemicals induced a significant increase in MN frequencies in binucleated (BN) cells (P < 0.001). The maximal effect was reached at a dose of 0.50 microgram/ml. At this dose, VRB produced an approximately 5 fold increase with respect to the control frequency of MN, while with VCR, this frequency increased 10-fold. Both drugs produced a slowing of the cell cycle, causing a decrease in the percentage of BN cells. This effect was lower with VRB. The percentages of centromere-positive MN were 89.79 and 87.60% in VRB- and VCR treated cultures, respectively (control 27.03%). The high percentage of positive signals in treated cultures (P < 0.001) indicates that the MN contained whole chromosomes. Our results confirm the aneugenic mode of action of these chemicals, VRB having less genetic effect. PMID- 10474824 TI - In vivo dominant lethal effect of pyrimethamine in male mouse germ cells. AB - Pyrimethamine is used for treatment of malaria and toxoplasmosis. The embryotoxicity and clastogenicity of pyrimethamine is known and our aim was to investigate its dominant lethal effect in vivo. For this purpose, we used three groups of Swiss-albino male mice and a control group. We injected males with doses of 16, 32 or 64 mg/kg pyrimethamine and housed them with 10 females/male for each mating interval. Females were sacrificed and their uteri were evaluated for dominant lethality. As a result of this study we found that pyrimethamine induced dominant lethal mutations in the third, fourth and sixth weeks at the 64 mg/kg dose level, without the effect being dose-dependent. We conclude that pyrimethamine is a suspected germ cell mutagen. PMID- 10474825 TI - Human sperm DNA integrity assessed by the Comet and ELISA assays. AB - DNA integrity in sperm is essential for the accurate transmission of genetic information and therefore the maintenance of good health in future generations. The ELISA and Comet assays, two techniques that detect DNA damage in cells, are compared in this study of DNA integrity in human sperm. Both techniques rely on alkaline unwinding for the release of single strands of DNA from the nucleus. The ELISA detects single strands immunochemically whereas the Comet assay measures single strands drawn out by electrophoresis, stained with ethidium bromide and quantified by image analysis. The two techniques, both modified for use with sperm, detect similar levels of baseline DNA damage along with similar dose dependent patterns of induced damage by X-ray irradiation at 10 and 30 Gy (P < 0.05). The assays are also comparable in the detection of a significant protective effect by ascorbic acid (300 and 600 microM) and alpha-tocopherol (30 and 60 microM) on DNA integrity, both at baseline levels and following X-ray irradiation (p < 0.01). The advantages and disadvantages of each technique are discussed. PMID- 10474826 TI - Detection of the '4977 bp' mitochondrial DNA deletion in human atherosclerotic lesions. AB - The presence of the 4977 bp deletion ('common deletion') in the mitochondrial DNA (mtDNA) is associated with defects in the metabolic machinery acquired during ageing as a hallmark of a degenerative phenotype. We analysed 27 samples (18 from surgical patients and nine from autopsy cases) of DNA extracted from smooth muscle cells of abdominal aorta fragments affected by atherosclerotic lesions. The deletion was detected by PCR amplification gel electrophoresis and characterized by sequencing of the PCR product. The mtDNA 'common deletion' was detected in all analysed samples. However, its levels were not particularly high, which may be ascribed to the fact that smooth muscle cells in atherosclerotic lesions have a lower energy requirement and an appreciable proliferation rate, as compared for instance with cardiac myocytes. When the subjects were divided into two numerically equivalent age classes (60-72 years plus a 45-year-old subject versus 73-95 years), the deletion had significantly higher levels in the older subjects. Conversely, its presence did not correlate with source (surgical or autoptic), sex, cigarettes consumption, other clinical and anamnestic parameters or with the levels of adducts and 8-hydroxy-2'-deoxyguanosine measured in the nuclear DNA of the same samples. A previously unreported deletion of 5111 bp was additionally found in the mtDNA from a 45-year-old woman. The origin of this lesion seems to be compatible with the slipped mispairing model proposed for the 'common deletion'. PMID- 10474827 TI - Chromosome breakage at sites of oncogenes in a population accidentally exposed to radioactive chemical pollution. AB - The purpose of the present study was to investigate the level of aberrations at fragile sites of chromosomes in peripheral blood lymphocytes of the population of an area polluted with radionuclides, following an accident at the Siberian Chemical Plant. We carried out the micronucleus test to screen people with radiation-related cytogenetic effects. Of the 1246 inhabitants of the settlement of Samus examined, 148 showed a significantly increased frequency of micronucleated erythrocytes and were selected for chromosome analysis as a radiation-exposed group. Additional analysis was carried out on 40 patients with gastric cancer and atrophic gastritis with stage II-III epithelial dysplasia. Eighty six individuals from a non-polluted area were used as a control group. Chromosomal breaks and exchanges occurred preferentially in chromosomes 3 and 6 among radiation-exposed persons and patients. The regions 3p14-25 and 6p23 were damaged most often. There was a tendency to preferential involvement of q21-25 of chromosome 6 in patients with gastric cancer and atrophic gastritis. Specific damage at certain chromosome sites was observed in the radiation-exposed population as well as in patients with gastric cancer. Most often this damage was located near oncogene loci, which could imply that chromosome damage induced by radiation is likely to be a predisposing factor to the expression of oncogenes and malignant transformation of cells in exposed individuals. PMID- 10474828 TI - A comparison of bleomycin-induced damage in lymphocytes and primary oral fibroblasts and keratinocytes in 30 subjects. AB - The number of chromatid breaks in peripheral blood lymphocytes (PBL) after exposure to bleomycin in the S/G2 phase of the cell cycle (in the literature referred to as 'mutagen sensitivity') is associated with an increased risk of environmentally related cancers, including oral cancer. The aim of this study was to elucidate whether mutagen sensitivity measured in lymphocytes actually reflects chromosomal instability of normal cells in the areas in which tumors develop. Therefore, bleomycin-induced chromosomal damage in and growth inhibition of cultured oral fibroblasts and oral keratinocytes from 30 persons were compared with the standard mutagen sensitivity score in PBL. A correlation was found for the percentage of aberrant metaphases between PBL and oral fibroblasts but not for the number of breaks per cell. These data do not allow a conclusion to be drawn on the use of fibroblasts to study cancer risk. Within the fibroblasts it was found that a high number of breaks per cell was associated with less growth inhibition, indicative of damage-resistant growth. Oral keratinocytes were extremely sensitive to bleomycin, as indicated by a strong cell cycle block which resulted in a mitotic index too low to determine chromosomal breaks. Moreover, in the cell proliferation assay keratinocytes were found to be 100 times more sensitive as compared with fibroblasts. There was no correlation between bleomycin sensitivity of keratinocytes compared with fibroblasts from a single patient as measured by growth inhibition. This may be due to the strong influence of alcohol consumption by the subjects, which was found to increase the sensitivity of keratinocytes but not of PBL and fibroblasts. In conclusion, oral fibroblasts but not keratinocytes can be used to measure sensitivity for chromatid breaks. The apparent influence of environmental factors on keratinocytes makes them a useful source to study exposure characteristics but limits their application for the determination of genetic factors. PMID- 10474829 TI - Hydrogen peroxide and coffee induce G:C-->T:A transversions in the lacI gene of catalase-defective Escherichia coli. AB - The mutagenicity of hydrogen peroxide (H2O2) was compared with that of coffee, a complex mixture which generates H2O2. An Escherichia coli strain defective in catalase activity (katG katE double mutant) and carrying a single copy mucAB (pRW144) plasmid was constructed to enhance the mutagenic response to oxidants. The ability of the mucAB genes to influence the type, frequency and distribution of H2O2-induced mutations was also investigated in isogenic bacteria lacking pRW144. Induced mutational spectra were characterized and compared with that of spontaneous mutagenesis. A total of 444 independent forward mutations affecting the first 210 bp of the lacI gene were identified by DNA sequence analysis. The spontaneous mutation spectrum showed no bias (P = 0.52) for substitutions at G:C base pairs. In contrast, in the H2O2-induced spectrum substitutions occurred preferentially at G:C base pairs (P < 0.0001) with a preponderance of G:C-->T:A transversions (43.4% of H2O2-induced mutants versus 17.3% of spontaneous mutants). These data support the view that 7,8-dihydro-8-oxoguanine is the main premutagenic lesion induced by H2O2 and that catalase-defective bacteria have elevated levels of 8-oxoguanine in chromosome DNA after H2O2 exposure. Coffee produced a similar distribution of mutational events as H2O2 (P > 0.05), suggesting that this compound may be the main cause of the coffee-induced mutagenesis. The presence of plasmid pRW144 did not affect the frequency of H2O2 induced G:C-->T:A transversions, but caused an increase in A:T-->T:A transversions and a decrease in -1 base frameshifts. Although the frequencies of G:C-->T:A transversions were similar in all three induced spectra (H2O2 and coffee +/- pRW144), differences were observed in location of mutations throughout the target gene. PMID- 10474830 TI - The involvement of chromatin condensation in camptothecin-induced chromosome breaks in G0 human lymphocytes. AB - In the present study we evaluated campthotecin (CPT)-induced chromosomal damage in human lymphocytes in the G0 phase of the cell cycle as revealed by the premature chromosome condensation technique. The results obtained here indicate that CPT was able to induce chromosome fragments in the G0 phase of the cell cycle of human lymphocytes as detected in prematurely condensed chromosomes. This result appears to be rather surprising, since the DNA lesions produced by CPT (e.g. 'protein concealed' DNA single-strand breaks) should not produce any damage in G0. A possible explanation for this result could come from much evidence to suggest that chromatin condensation processes are significantly involved in the conversion of DNA lesions into chromosome breaks in prematurely condensed chromosomes. The unexpected clastogenic behaviour of CPT can be explained taking into account the chromosome condensation induced by mitosis promoting factors when human lymphocytes are fused in G0, thus converting the 'protein concealed' DNA single-strand breaks induced by CPT into chromosome breaks. The same perspective should be taken into consideration for breaks induced by CPT under normal physiological conditions in the G2 phase of the cell cycle. PMID- 10474831 TI - Inhibitory action of melatonin on H2O2- and cyclophosphamide-induced DNA damage. AB - Melatonin, the pineal gland hormone known for its ability to modulate circadian rhythm, has recently been studied in its several functions. It is believed to inhibit cancer growth, to stimulate the immune system and to act as an antioxidant. In particular, this latter activity is ascribed to two different mechanisms: stimulation of radical detoxifying enzymes and scavenging of free radicals. We used this compound in mammalian cells in vitro to investigate its mechanism of action in modulating DNA damage. Cytogenetic and cytofluorimetric analyses were performed. We show that melatonin is able to modulate chromosome damage (chromosomal aberrations and sister chromatid exchanges) induced by cyclophosphamide. Conversely, its involvement in modulating oxidative processes, thereby reducing DNA damage, is less clear. In particular, melatonin is able to decrease H2O2-induced chromosomal aberrations but not sister chromatid exchanges and has been found to induce oxygen species in a cytofluorimetric test (DCFH assay). PMID- 10474832 TI - A comparison of the effects of diverse mutagens at the lacZ transgene and Dlb-1 locus in vivo. AB - Transgenic assays permit the detection of mutations in any tissue, whereas endogenous mutations can be measured in very few. For this reason comparisons between these loci when both can be measured in the same cells are of considerable interest. Previous comparisons have been inconsistent: usually these loci have responded alike, however, in some cases the endogenous locus has been more sensitive and at other times the transgenic locus has been more sensitive. Here we report a comparison of the lacZ transgene of the MutaMouse and the endogenous Dlb-1 gene in the epithelium of the small intestine after acute exposure to seven mutagens. Benzo[a]pyrene, 5-bromo-2'-deoxyuridine, methyl methane sulphonate, ethyl methane sulphonate, N-ethyl-N-nitrosourea, mitomycin C and N-methyl-N-nitrosourea were all given by gavage to F1 (MutaMouse x SWR) mice. Mutations were quantified 2 weeks after the end of treatment. The data shows that all of the agents induced similar mutant frequencies at the Dlb-1 locus and at the lacZ transgene. The acute treatments generally produced only modest increases in mutant frequency at both loci. The higher background frequency observed at the lacZ transgene reduces the ability of the transgenic assay to detect the same absolute increase in mutant frequency. PMID- 10474833 TI - FISH analysis of 1cen-1q12 breakage, chromosome 1 numerical abnormalities and centromeric content of micronuclei in buccal cells from thyroid cancer and hyperthyroidism patients treated with radioactive iodine. AB - One of the health consequences of the Chernobyl nuclear power plant accident was a radioactive iodine-related increase in the incidence of thyroid cancer in exposed children. This radioisotope is used in the treatment of thyroid cancer and hyperthyroidism patients providing a convenient opportunity to study cytogenetic damage induced by known doses of radioactive iodine in treated patients. We used pancentromeric FISH on micronuclei and chromosome 1 tandem labelling FISH to monitor overall chromosome breakage and loss, 1q12 breakage and decondensation and chromosome 1 numerical abnormalities in buccal cells from 31 radioactive iodine-exposed hyperthyroidism and thyroid cancer patients. The overall outcome of the study, with 250,000 buccal cells analysed, is that there was no radioactive iodine-related increase in the frequency of micronuclei, 1q12 breakage, 1q12 decondensation or chromosome 1 numerical abnormalities. In addition, neither age nor gender, health status nor radioactive iodine dose modulated the frequency of the above cytogenetic end points. Although several uncertainties of these emerging molecular cytogenetic methodologies will require further experimentation, we conclude that, at the reported exposure levels, radioactive iodine did not induce detectable chromosome damage in buccal cells from treated patients. PMID- 10474834 TI - Effect of Tn10/Tn5 transposons on the survival and mutation frequency of halogen light-irradiated AB1157 Escherichia coli K-12. AB - We show here that the Tn10/Tn5 transposon when inserted into the chromosome of strain AB1157 makes the bacteria more sensitive to and less mutable by halogen light irradiation. These effects are most probably caused by depletion of UmuD and UmuC proteins since: (i) transformation of the transposon-bearing bacteria with plasmids harbouring umuD'C (or umuDC) leads to recovery of the original survival and mutation frequencies; (ii) insertion of Tn10/Tn5 into chromosomal DNA has no effect on the level of mutation induced by ethyl methane-sulphonate treatment, a mutagen whose activity is umuDC-independent; (iii) the decline in survival is in about the same range for Tn10-bearing bacteria as for bacteria with deleted umuDC. However, whereas transformation of bacteria deleted in umuDC with plasmids carrying umuD'C/umuDC leads to full recovery of halogen light induced mutability, recovery of survival is poor. This suggests that the mechanisms leading to umuDC-dependent mutagenesis and umuDC-dependent protection of cell survival are different. None of these effects occurs in bacteria bearing the Tn9 transposon in their DNA. PMID- 10474836 TI - Transgenic mouse mutation assay systems can play an important role in regulatory mutagenicity testing in vivo for the detection of site-of-contact mutagens. AB - Transgenic mouse mutation assays, such as MutaMouse (lacZ, CD2F1) and Big Blue (lacI, B6C3F1), afford the opportunity to evaluate the mutagenic potential of chemicals in any target organ in vivo. This paper discusses published data collected from the analysis of the skin, stomach and lung DNA after topical, oral and inhalation exposure, respectively. These data indicate that both MutaMouse and Big Blue should play an important part in the evaluation of genotoxicity in vivo, particularly where the endpoint or target tissue available in the more conventional tests is inappropriate. It is concluded that there is a distinct role for this type of assay in genetic toxicology testing. For substances applied to the skin or dosed orally or by inhalation and which are unlikely to reach either the bone marrow or the liver, then data derived from these assays may be more relevant to an assessment of possible risk to man than the currently used unscheduled DNA synthesis in liver and cytogenetics assays in bone marrow or peripheral blood. PMID- 10474835 TI - Mutations at G:C base pairs predominate after replication of peroxyl radical damaged pSP189 plasmids in human cells. AB - The mutagenicity of peroxyl radicals, important participants in lipid peroxidation cascades, was investigated using a plasmid-based mutational assay system. Double-stranded pSP189 plasmids were incubated with a range of concentrations of the water-soluble peroxyl radical generator 2,2'-azobis(2 amidinopropane) hydrochloride (AAPH). Following replication in human Ad293 cells, the plasmids were screened for supF mutations in indicator bacteria. Exposure to peroxyl radicals caused strand nicking and a decrease in transfection efficiency, which was accompanied by a significant increase in supF mutants. Each of these effects was abolished in the presence of the water-soluble vitamin E analogue Trolox. Automated sequencing of 76 AAPH-induced mutant plasmids revealed that substitutions at G:C base pairs were the most common changes, accounting for 85.5% of all identified mutations. Of these, most comprised G:C-->T:A transversions (53.5%), with lesser contributions by G:C-->A:T transitions (23.9%) and G:C-->C:G transversions (22.5%). Collectively, these data confirm our previous findings concerning the spectrum of mutations produced upon bacterial replication of peroxyl radical-damaged phage DNA and extend them by showing that such damage has mutagenic consequences during replication in more complex eukaryotic systems. PMID- 10474837 TI - [Stormy and unpredictable. Surgical treatment of lumbar intervertebral disk displacement]. PMID- 10474838 TI - [Surgical approach in lumbar intervertebral disk displacement. Topographical principles and characteristics]. AB - For the operative procedure in lumbar disc extrusion it is important to expose the prolapsed disc and sequestrum without disintegration of the musculature and branches of r. dorsalis nervi spinalis or provoking lesions in the spinal canal. A classification helps to determine exact description of localization of disc material in craniocaudal and mediolateral directions. It can be correlated with the interlaminar window as a structure seen in the operative procedure. A speculum helps to limit the exposure for an interlaminar as well as a lateral access, thus reaching the structures concerned in the spinal canal with minimal irritation of the surrounding tissues. For interlaminar access partial excision of the lig. flavum is sufficient. The extrusions in the foraminal region that can be exposed laterally from a medial skin incision, as well as from a paramedial are between m. multifidus and m. longissimus. In revision surgery, the upper laminar corner is advantageous for entering the spinal canal. PMID- 10474839 TI - [Surgical treatment of lumbar intervertebral disk displacement. Indications and methods]. AB - The decision about operative or non-operative treatment of herniated lumbar discs has to be based on the results of clinical and neurological examination. Results of imaging techniques as CT or MRI are secondary. Various percutaneous endoscopic or open techniques have been described. The most appropriate technique is determined by the anatomical situation of the dislocated lumbar disc tissue. Intradiscal percutaneous therapies such as percutaneous nucleotomy and laser decompression are possible, but they revealed poor results in prospective randomized and controlled studies. Chemonucleolysis remains as an effective intradiscal technique at the time, but is afflicted with side effects and complications. Endoscopic transforaminal or interlaminary techniques are still experimental. Whenever open surgery is necessary, microsurgical techniques are preferred. If a certain technique is to be applied, there has to be the correct indication, technical support for and experience in using the choosen method. PMID- 10474840 TI - [Open lumbar intervertebral disk operation. Technique and results]. AB - Various operative techniques are available for the treatment of lumbar disc herniation. The conventional lumbar discotomy is currently considered to be the standard procedure. Several large statistical studies have shown that the results in the majority of cases are very good and the complication rate is low if it is carried out correctly. A particular advantage of this technique is that almost all indications can be handled with a procedure that can be extended if necessary. Despite the availability of newer refined techniques (microdiscectomy or percutaneous procedures), the open lumbar discotomy still has an outstanding role in the management of lumbar disc herniation. Failure results more frequently from false indications or inadequate preoperative planning and less from technical variations. PMID- 10474841 TI - [Microsurgical lumbar intervertebral disk operation. Technique and complications]. AB - Lumbar Microdiscetomy requires special instruments and provides a good view with a narrow approach. The microscope is the best surgical aid. Alternatives are a retractorintigrated light or headlamp. A specific complication of lumbar microdiscectomy--the wrong level--can be minimized by an exact preoperative planning. Other complications like dural lesions and exessive bleeding are less frequent with the microscope because of the better view. The most severe complications that can occur with any lumbar disc operation, major vessel or visceral injury, can be avoided by lusing the new rongeur with a depth guard. PMID- 10474842 TI - [Percutaneous nucleotomy procedures in lumbar intervertebral disk displacement. Current status]. AB - The discussion on the results of open spinal surgery, in particular, on complications as a result of open surgery, e.g., the so-called post-discotomy syndrome, has led to the development of less invasive methods, especially for treatment of lumbar disc prolapses. Percutaneous nucleotomy procedures are based on a variety of intradiscal diagnostic and therapy methods, such as discography, retroperitoneal spinal disc fenestration or chemonucleolysis. Since the 1970s, various methods such as mechanical percutaneous nucleotomy, automated percutaneous nucleotomy, intradiscal laser procedures and, in part, endoscopic intradiscal procedures have been developed and also used clinically. The clinical results vary from 30% good/very good results to almost 100% good/very good results, depending on the author and study. It is obvious that not many prospective randomized studies using either conservatively or surgically treated control groups have been conducted. The results of these studies on intradiscal therapy methods are often analyzed more critically and with lower success rates than the partially retrospective stuides of a single method, which often have large case numbers, but do not always meet the demanding criteria for scientific study design. To attain good results, percutaneous nucleotomy procedures can only be used to treat select cases of lumbar disc disorders according to strict indication criteria. At the present time it can be concluded that, in comparison to endoscopic und microsurgical methods, percutaneous nucleotomy procedures must be restricted to a small number of cases. PMID- 10474843 TI - [Chemonucleolysis. Development, experiences, prospects]. AB - Concerning the so-called minimal invasive procedures currently available for the treatment of lumbar disc prolapse (percutaneous endoscopic discectomy, APLD, laser decompression, chemonucleolysis), the intradiscal application of Chymopapain represents the method with the longest period of clinical use and experience. Long-term studies have shown good clinical results. When considering of the indication and the few contraindications--particularly allergic diathesis- chemonucleolysis provides a low-risk, efficient, minimally invasive therapy that closes the therapeutic window between conservative and open surgical treatment. PMID- 10474844 TI - [Transforaminal endoscopic microdiscectomy. Indications and short-term to intermediate-term results]. AB - 101 patients with lumbar disc herniation were treated by transforaminal endoscopic microdiscectomy between 5/94 and 6/97. Caused by technical problems the procedure could not be successfully completed in 3 patients. They must be excluded from the presented study as 9 others, who were lost for follow up. So 89 patients were followed with a median follow-up time of 28 months (15-56 months). 69 patients (78%) were satisfied. 16 patients needed a second operation. In 11 of these 16 patients it was done by an open procedure, in 5 by transforaminal endoscopic microdiscectomy again. 4 of these 5 patients showed a good or satisfying result. As in open procedures the most important prognostic factor is a promptly disappearing radicular leg pain. Only 6 of 61 patients with a radicular leg pain disappearing within one week were reoperated, but 10 of 28, who showed a leg pain persisting longer than one week postoperatively. PMID- 10474845 TI - [Neuromuscular scoliosis]. PMID- 10474846 TI - Hypometabolism in the limbic system of cancer patients observed by positron emission tomography. AB - Brain images obtained by a positron emission tomography with 18F fluorodeoxyglucose from 19 pretreatment cancer patients with variable cancers except brain cancers were compared with those from 17 inpatients with benign diseases. Relative reduction in regional cerebral metabolism of glucose was found mainly in the limbic structures including the bilateral orbitofrontal cortex, lateral frontal cortex, cingulate gyrus, thalamus, hippocampus and parahippocampal gyrus, insulae, and basal ganglia. The regional metabolism tended to be low in the patients who had severe pains which required opiates and morphines, and tended to be preserved in the patients who were already disclosed of their malignancy based on previous studies such as biopsy. Our findings would support that psychological deficits in cancer patients are associated with abnormalities in regional brain metabolism in the limbic system. PMID- 10474847 TI - Gender as a predictor of burden and psychological distress of elderly husbands and wives of cancer patients. AB - The aim of the study is to explore whether gender is a cause of differences in the perception of burden on the part of 39 elderly wife caregivers and 21 elderly husband caregivers of cancer patients. The patients were being treated at an oncological medical center in northern Israel. At the time of interview the patients had no evidence of the disease. Stepwise regression analysis indicated that caregiver gender (i.e., husbands) and age, as well as the patient's psychological distress, predicted greater burden, although gender did not predict greater psychological distress. PMID- 10474848 TI - A group cognitive behaviour therapy programme with metastatic breast cancer patients. AB - One-hundred and twenty-four patients with metastatic breast cancer were randomised to either a group Cognitive Behaviour Therapy (CBT) intervention, or to a no-therapy control group condition. Both groups received standard oncological care; however, therapy recipients also attended eight weekly sessions of group CBT, followed by a family night, and three further monthly sessions. Patients completed the 'Profile of Mood States' (POMS) and the Coopersmith Self esteem Inventory (CSI) before and after therapy, and at 3 and 6 month follow-up periods. Outcome data in the period following therapy showed reduced depression and total mood disturbance, as well as improved self-esteem amongst therapy participants, relative to a no-therapy control group. These improvements were no longer evident at the 3 or 6 month follow-up assessments. We also report on the difficulties associated with conducting a group intervention with this patient cohort. PMID- 10474849 TI - Description of a group Cognitive Behaviour Therapy programme with cancer patients. AB - Cognitive Behaviour Therapy (CBT) has already been shown to be highly effective in the treatment of various psychological disorders within mental health populations; however, it has not been widely tested in the treatment of cancer patients. In the last decade there has been growing interest in the application of CBT interventions within psycho-oncology, and some studies have reported on its efficacy, both with individuals and in group-therapy contexts. To date there have been few descriptions of how a CBT programme for cancer patient groups can be structured and delivered. We present a description of a 12-session CBT programme that has been developed by our centre, for delivery to cancer patient groups. PMID- 10474850 TI - Depression and cancer: an appraisal of the literature for prevalence, detection, and practice guideline development for psychological interventions. AB - This paper reviews the current literature concerning the prevalence and incidence of depression in the general population and within the population of people living with cancer where depression is defined using current standards and guidelines. Reviews of recent work where the treatment of depression was the focus of study are also presented. Finally, studies that have specifically addressed the psychological treatment of depression among cancer patients are critically reviewed. Suggestions for practice guideline development of psychological interventions are offered based on the review. PMID- 10474851 TI - A systems model for adaptation to somatic distress among cancer survivors. AB - Applied coping research has generally failed to fulfill its goal of providing an empirical basis for clinical interventions, and research on coping with cancer is no exception. This can be attributed in large measure to the wide gap between coping theory and coping research. Theories of stress and adaptation are complex systems formulations that present conceptual and methodological challenges and thus make testing comprehensive models difficult. The present paper reviews arguments for a microanalytic strategy through which researchers can increase coverage of relevant variables from broad systems models of stress and coping by concentrating their resources on selected, high-frequency, high-stress problems. The utility of this approach for formulating problem-specific systems models is illustrated using the example of coping with somatic distress among cancer survivors. PMID- 10474852 TI - An evaluation of a psychosocial intervention for survivors of childhood cancer: paradoxical effects of response shift over time. AB - This study examined the impact of response shift on a psychosocial treatment evaluation of 22 young adult cancer survivors. An age-matched cohort of 54 healthy controls were included in the study to provide a comparison for normative levels and structure of quality-of-life (QOL). It was found that this evaluation of a psychosocial intervention for young adult cancer survivors was notably influenced by response shift phenomenon. Standard analyses suggested that the intervention had no impact on measured aspects of well-being. It did appear to yield an immediate gain in reported global QOL, but seemed to cause a significant decline over time. By considering response shift, it was highlighted that an apparently deleterious effect on QOL was largely a function of response shift. This response shift effect was reflected not only in changes in internal standards, but also in values and in conceptualization of QOL. The intervention seemed to have normalized survivors' conceptualization of QOL so that it was increasingly similar to their age-matched cohort. Future psychosocial intervention research should explicitly consider response shift in a randomized treatment evaluation. PMID- 10474853 TI - Development of quality of life questionnaire in Japan: quality of life assessment of cancer patients receiving chemotherapy. AB - The Japanese Quality of Life Research Group has developed a general questionnaire suitable for assessing the Quality of Life (QOL) in patients undergoing chemotherapy. The questionnaire covers four major categories: (1) daily activities, (2) physical condition, (3) social activities, and (4) mental and psychological status. The State-Trait Anxiety Inventory (STAI), Self-Rating Depression Scale (SDS), and Performance Status (PS) were used as external measures of quality of life and for the validation of our tool. On the basis of two basic surveys and two studies we selected 22 questions from a larger set of items. Validity and reliability were verified for the final 22-question form. This questionnaire, named the QOL Questionnaire for Cancer Patients Treated with Anticancer Drugs (QOL-ACD), can be used to detect changes in QOL over time. Its use as an additional outcome measure in Phase III chemotherapy trials should be encouraged. PMID- 10474854 TI - A careful investigation of an important phenomenon. PMID- 10474855 TI - Clarification regarding comments about a hypothesis. PMID- 10474857 TI - [Gene frequencies and heterozygosity of the population of Donetsk Province, Ukraine by the alleles of the ABO and Rhesus systems]. AB - Frequency and heterozygosity indices of AB0 and Rh gene systems in the population of Donetsk Province were calculated. Uneven distribution of the genes was found and heterozygosity indices of the population were 0.554-0.573 for AB0 and 0.410 0.499 for Rh. Heterozygosity in this population was higher than average heterozygosity in total population of Ukraine as a result of intensive migrations and prevalence of heterolocal marriages over homolocal ones. PMID- 10474856 TI - [Corpus uteri cancer morbidity and a genetic-mathematical analysis of the proband's pedigrees]. AB - Steady increase of the number of women that primary fell ill with uterus cancer has noted in Ukraine in 1980-1994. The clinic-genealogical analysis of the 262 families with uterus cancer patients of the Kiev region was made. Multifactorial type of heredity predominates. Share in general tendency to malignant tumors of the genetic component was 45.64 +/- 7.20 and of the environment factors was 54.36 +/- 7.20. Closest relatives of probands with the uterus cancer have to be subjected to systematic inspection. PMID- 10474858 TI - [The mutational spectra of gene p53 in different types of tumors]. AB - The comparative analysis of the frequencies of nucleotide exchanges in mutational spectra of gene p53 (5-8 exons) between germline cancer-prone families (Li Fraumeni syndrome), between somatic mutations in the tumors of different histogenesis and cell lines, obtained from them, was carried out. The nucleotide positions with high level of mutation events (mutational "hot spots"), typical for germline mutational spectra, tissue-specific patterns of their disappearing and appearing of new ones in solid tumors in vivo, nearly complete absence of "hot spots" in lymphomas and cell lines in vitro were revealed. The obtained results allowed to suggest, that one of the leading factor controlling hot-spots distribution in 5-8 exons of gene p53 is the specificity of cell division conditions in vivo and in vitro. PMID- 10474859 TI - [An analysis of chimeric SMN genes--new possibilities in the study of the molecular genetic nature of mutations and in the diagnosis of spinal muscular atrophy (SMA)]. AB - Results of analysis of chimeric SMN genes among some high SMA-risk families from Ukraine using the EcoRV and DdeI restriction enzyme hydrolysis of PCR products is presented. Chimeric cen/telSMN gene was detected in probands with homozygous deletions of telSMN exon 7 only, as well in proband with absent of homozygous deletion of exons 7 and/or 8 of the SMN gene. Effectivity of approach of detection of chimeric SMN genes based on the EcoRV and DdeI restriction enzyme analysis of PCR products and mechanisms of formation of chimeric SMN genes are discussed. PMID- 10474861 TI - [The cytoradiography of the DNA and proteins in different types pf cells after the irradiation of rats with decimeter waves]. AB - Microwave (2450 MHz, 1 mW/cm2, continuous regime) influence on DNA and protein synthesis in bone marrow, testes and liver cells of Fisher-line rats at early stage of formation of defence-adaptation reactions was studied. Cytoradiographic analysis of tissue smears and prints using 3H-thimidine and 35S-methionine was made. Type of reaction to microwave irradiation depends on ploidy, rate of differentiation and functional activity of cells. PMID- 10474860 TI - [The cytospectrophotometric study of the DNA content in benign and malignant processes in the breast]. AB - Cytospectrophotometric analysis of DNA content in nuclei of the epithelial cells in fibroadenoma, fibroadenomatous and breast cancer revealed that mean value of DNA content in breast cancer is reliably higher than in benign processes in breast. Modal class in fibroadenoma and fibroadenomatous formed by di- and tetraploid cells, in breast cancer-tetraploid and higher depending on histological structure of tumor. Ratio of aneuploidy is higher in invasive ductal carcinoma and scirrhus [correction of skir] in comparison with invasive lobular carcinoma and other carcinomas, that evidences higher aggressiveness of these tumors. PMID- 10474868 TI - [Xenotransplantation]. PMID- 10474869 TI - [Parenteral and enteral nutrition in palliative medicine]. AB - Artificial nutritional support does not alter the natural course in patients with malignant disease. The outcome of these patients is mainly determined by the type and stage of the underlying tumor. Progress of the underlying disease is often paralleled by malnutrition which in turn facilitates complications and may reduce survival and quality of life. Nutritional support can be applied to maintain body weight, immune function and quality of life. Enteral nutrition can be applied with a functioning gastrointestinal tract and has been proven to be superior compared to parenteral nutrition. Maintenance of intestinal mucosal function due to enteral substrate application prevents disruption of intestinal barrier function as well as the overgrowth of intestinal microorganisms. Using a step-by step approach dietetic counselling in combination with augmented oral caloric intake should be the first measure. The next step to take is nutritional support by enteral tube feeding using formula diets. Parenteral feeding should only be used if other options to support caloric intake have failed. PMID- 10474870 TI - [Xenotransplantation. Possibilities of animal breeding]. AB - The pig is the most likely donor organism for xenotransplantation of organs to humans. However, since this constellation is discordant, hyperacute rejection needs to be overcome. This review summarises current strategies of genetically modifying pigs for xenotransplantation. Limitations of the classical method of DNA-microinjection and new perspectives arising from the possibility of cloning animals from cultured cells are discussed. PMID- 10474871 TI - [Mechanisms of xenotransplant rejection]. AB - The recent shortage of allogeneic human organs in transplantation medicine has led to a renewed interest in evaluating the feasibility of xenotransplantation, particularly of porcine origin. Discordant xenotransplants between phylogenetically distant species such as pig and primates are challenged by a series of strong rejection reactions. Hyperacute rejection is dominantly mediated by humoral responses of the immune system (natural antibodies, complement cascade) and the activation of coagulation factors, whereas delayed xenograft rejection and T-cell mediated responses are also mediated by elements of the cellular immune system. The mechanisms of chronic xenograft rejection remain to be elucidated. The present review is focused on the mechanisms of rejection and the immunobiology of discordant, vascularized xenotransplants. We attempt to provide a brief description of the dynamics, the humoral and cellular mediators, and the pathogenesis of xenograft rejection. Finally, an overview on potentially therapeutic strategies to overcome xenograft rejection is presented. PMID- 10474872 TI - [Risk of infection causes by viruses in xenotransplantation]. AB - The risk of transferring exogenous pig viruses to man during organ xenotransplantation can be controlled by keeping the pig donors pathogen-free. A risk of mobilizing pig endogenous retroviruses and a "xenozoonosis"-infection in human organ recipients cannot be excluded according to recently reported virological cell culture experiments. The present state of research, however, does not allow an answer to the question of whether or not a disease might be caused by such an infection and whether such a virus might be contagious to third persons. PMID- 10474873 TI - [Xenotransplantation of hDAF-transgenic swine hearts]. AB - Hearts of transgenic pigs expressing a human regulator of complement activation, decay accelerating factor (hDAF), were transplanted either heterotopically into the abdomen of cynomolgus monkeys or orthotopically into baboons. None of these transgenic hearts was hyperacutely rejected. Immunosuppression with a combination of cyclosporine A, cyclophosphamide and steroids produced a maximum survival of 62 days (median 40 days) in the heterotopic model. Transgenic hearts transplanted into the orthotopic position allowed a maximum survival of 9 days (median 2.5 days). A more effective and less toxic immunosuppressive protocol for the prevention of accelerated xenograft rejection is the subject of ongoing research. The use of organs from transgenic pigs may help to solve the problem of donor shortage in clinical allotransplantation. PMID- 10474874 TI - [Experimental xenogenic lung transplantation]. AB - Lung transplantation has become a valuable therapeutical option for patients with end stage vascular and parenchymal pulmonary disease. The main limitation is the donor organ shortage, which is for lung transplantation even more pronounced than for other solid organs. Therefore, the search for alternative sources in the field of xenotransplantation as potential bridging to human allograft transplantation has emerged over the last few years. Pulmonary xenotransplantation at this stage is still an experimental procedure. Different concordant and discordant models have been investigated for specific organ functions, but in subhuman models no long-term function could be achieved to date. The purpose of this article is to present recent results in pulmonary xenotransplantation models. PMID- 10474875 TI - [Human xenogenic kidney transplantation from the clinical viewpoint]. AB - Pig-to-primate transplantations of hDAF-transgenic kidneys achieved survival times of up to 70 days. Pilot projects for clinical kidney xenotransplantation seem to be possible in the near future. The problems of hyperacute rejection seem to be solved. Delayed xenograft rejection remains a serious problem. Further medical, ethical and practical issues have to be solved, before Phase-I-studies can be anticipated. In contrast to other organs, kidney xenotransplantation is associated with low operative risk. Xenotransplant dysfunction or rejection could be treated with conventional hemodialysis. PMID- 10474876 TI - [Xenotransplantation of the liver]. AB - The development of pigs transgenic for human regulators of complement activation resulted in the nearly total elimination of episodes of hyperacute rejection following discordant solid organ xenotransplantation. Following discordant heart or kidney transplantation, in subhuman primates, graft survival rates of up to several months can be observed. In contrast to these organs, the xenotransplantation of the liver is associated with the inherent problem of the immunological and metabolic compatibility of the large variety of xenoproteins generated. Based on a review of data mainly derived from experimental ex-vivo xenoliver perfusions in patients with hepatic coma, whole organ orthotopic or heterotopic liver xenotransplantation currently is not likely to become a relevant option for the treatment of patients with endstage liver failure. In contrast, clinical studies utilizing different forms of bioartificial liver assist devices are currently underway. Based on preliminary data published, this form of liver support therapy might enter the clinic in the near future. PMID- 10474877 TI - [Transplantation of porcine Langerhans islets for therapy of type I diabetes. The way to clinical application]. AB - Transplantation of isolated pancreatic islets provides an interesting alternative to the present cure for diabetes: insulin injections and pumps. These are characterized by an insufficient glucose haemostasis and in the long run can induce kidney failure, blindness, heart failure, and amputations. Up to now more than 293 allogeneic islet transplantations have been performed in diabetics with chronical kidney failure. Despite some success, no real breakthrough has been yet achieved, though great efforts are being made to improve the various methodological steps on the way to clinical transplantation. The use of animal (xenogeneic) organs could be a solution to overcome the shortage of allogeneic donors. The current experimental and clinical research focuses on the use of pigs as organ donors, which have a number of advantages over the immunologically more compatible primates. This article reports on success and open questions concerning the efforts to isolate porcine islets for future clinical transplantation: the search for a suitable pig breed, the various isolation steps, purification and in vitro culture, transplantation models using-small and large animals, first clinical trials, and immunological reactions against the xenogeneic tissue. In addition, strategies to circumvent tissue rejection and future perspectives are discussed. PMID- 10474878 TI - [Xenotransplantation from the ethical viewpoint. An outline]. AB - The duty to save and to preserve lives on one hand, and the scarcity of human donor organs on the other hand call for a search for new organ sources, including xenogenic ones. Xenotransplantation, however, is not only in need of medical research, but also of ethical analysis. The latter is not to be considered a substitute for moral intuition, but rather a foundation of it by way of a critical evaluation of the ethical principles and reasons involved. This basically demands an analysis of the legitimacy of the aims and of the acceptability of the means for xenotransplantation. It includes safeguarding informed consent; risk assessment and the protection of not only the recipient, but also others; the question of limitation of personal rights; allocation problems; and last but not least animal protection. The aim is to clarify the ethical status of xenotransplantation in general and the question of a moratorium regarding clinical trials due to unsolved problems of infectivity and immunosuppression in particular by way of an integrative approach to both scientific developments and ethical analysis. PMID- 10474879 TI - [Identifying the recurrent laryngeal nerve by intraoperative neuromonitoring]. AB - 301 patients underwent thyroid surgery in 1998 by using the intraoperative neuromonitoring. The documentation was done prospectively. The system of intraoperative neuromonitoring consists of a stimulation circuit and an electromyographic record. We placed the deriving electrode transligamentally through the cricothyroid membrane. By relaxometry we investigated the influence of the relaxation level on the electromyographic record of the vocal muscle. Using a standardized operative technique we found a side-related rate of primary palsy of 2.3% (n = 13), from which 61.5% (n = 8) showed to be only temporary during a postoperative follow-up period. The intraoperative neuromonitoring with the purpose of identification of the recurrent laryngeal nerve is a safe and reliable method. PMID- 10474880 TI - [Significance of combined anterior cruciate ligament and meniscus injury]. AB - The combined lesion of the anterior cruciate ligament (ACL) with simultaneous or secondary meniscal tears has severe consequences for the knee joint. By a synopsis of the literature and own experimental investigations in sheep it can be shown that in ACL deficient knees secondary meniscal lesions occur, what finally ends in osteoarthrosis. The combined lesion of the ACL and the meniscus results in a more severe damage of the joint than the sum of the single lesions. ACL reconstruction in these cases is very important, because meniscal tears cannot heal in unstable knees. On the other hand resections of the menisci have to be avoided, because the loss of these secondary stabilizers and synergists of the ACL increases the risk of joint degeneration. PMID- 10474881 TI - [From Murphy's button to the Valtrac Ring. 100 years in search of a paradigm]. AB - In the context of continuous development in the area of anastomotic sutures leading to exceptional results in surgical practice, the biofragmentable anastomosis ring (BAR) described in 1985 by Hardy et al. represents a breakthrough in a 100 years' search of a paradigm. The first anastomotic button created in 1892 by J.B. Murphy was at once accepted as a quick and safe method of intestinal anastomosis. In 1896 Czerny demanded the following: "The task of technology is ... to create buttons with material that is entirely or partly dissolved in the intestinal lumen." Polyglycolic acid--developed in the sixties and now in widespread use for resorbable surgical sutures--was the material to fulfill the requirements already stated in the relevant literature 100 years ago, namely in the form of Hardy's BAR, which represents a redesigned Murphy button exploiting the recent biotechnological developments of this century. PMID- 10474882 TI - [Transabdominal preperitoneal hernia repair (TAPP). Results of 1,000 completed operations]. AB - It was the aim of this report to evaluate the laparoscopic transabdominal preperitoneal hernia repair (TAPP) which has been standardized at our department. Along with the demographic characterisation of 795 patients with 1000 inguinal hernia repairs we report about complications and early recurrences. The patient data were collected prospectively. The rate of follow-up amounted to 79.9% with an average follow-up of 1 year. The complications were divided into intraoperative, minor, major, as well as severe ones. In 30 repairs minor complications (3%) were detected. Major (n = 28) and severe (n = 9) complications were detected in 3.7% of the cases. There were two deaths, 3 patients with an intestinal obstruction due to adhesions (2 segmental small bowel resections), two patients with testicular atrophy, two mesh infections, two trocar hernias, 6 surgical revisions for removal of hematomas, one exploration of a testicle, 4 diagnostic laparoscopies for suspected recurrences with a negative result, and 15 patients with a nerve irritation syndrome. The early rate of recurrence was 0.7%. In 6 cases primary hernias had been repaired and in one case a recurrent hernia. The recurrent hernias became apparent in an average of 2 years (minimum 2.5 months, maximum 36 months) after surgery. The results of the clinical study demonstrate an acceptable rate of complications and a low rate of early recurrences. Based on these data we recommend the laparoscopic transabdominal preperitoneal technique and see an ideal indication in the case of bilateral, recurrent and femoral hernias. PMID- 10474883 TI - [Report on the 75th Congress of the Society of Bavarian Surgeons e.V. 23-25 July 1998 in Erlangen]. PMID- 10474884 TI - [Stapler hemorrhoidectomy--a new alternative to conventional methods]. PMID- 10474885 TI - [Intensive care medicine in surgery. General principles and special surgical intensive care medicine (1)]. PMID- 10474886 TI - The role of sentence context in accessing partial knowledge of word meanings. AB - Two experiments are reported that test the hypothesis that a reader can make use of the size of the semantic domain activated by a sentence context when inferring the meaning of a partially known word. We investigated words at three levels of knowledge: known, frontier, and unknown (e.g., Durso & Shore, 1991). Experiment 1 demonstrated that participants have knowledge about the meanings of words that they deny are part of the language (the unknown level), and that they make use of relative differences in the size of the semantic domains tapped by two sentences when asked to decide on correct usage of these unknown words. Experiment 2 demonstrated that participants have knowledge about the general semantic constraints operating on their unknown words, even when relative differences in size of semantic domains are controlled. Implications for the role of contextual constraints in vocabulary acquisition are discussed. PMID- 10474887 TI - American English usage frequencies for noun phrase and tensed sentence complement taking verbs. AB - The present research documents the American English usage frequencies for 136 verbs that occur with noun phrase and tensed sentence complements (e.g., accepted, noun phrase complement: The student knew the answer yesterday, tensed sentence complement: The student knew the answer was correct). This class of verbs has been the focus of numerous empirical studies investigating how syntactic ambiguity is resolved during language comprehension (Ferreira & Henderson, 1990; Garnsey, Pearlmutter, Myers, & Lotocky, 1997; Holmes, Stowe, & Cupples, 1989; Kennedy, Murray, Jennings, & Reid, 1989; Kennison, 1995; Rayner & Frazier, 1987; Trueswell, Tanenhaus, & Kello, 1993). The frequencies for noun phrase complements, tensed sentence complements, prepositional phrase complements, infinitival complements, and other usages are provided, as well as the frequencies with which specific verbs occur with the overt complementizer that in tensed sentence complements. PMID- 10474888 TI - Effect of aging on the monochromatic aberrations of the human eye. AB - We measured the contrast sensitivity (CS) of a group of older subjects through natural pupils and compared the results with those from a group of younger subjects. We also measured each subject's monochromatic ocular wave-front aberrations using a crossed-cylinder aberroscope and calculated their modulation transfer functions (MTF's) and root-mean-squared (RMS) wave-front aberrations for fixed pupil diameters of 4 mm and 6 mm and for a natural pupil diameter. The CS at a natural pupil diameter and the MTF computed for a fixed pupil diameter were found to be significantly poorer for the older group than for the younger group. However, the older group showed very similar MTF's and significantly smaller RMS wave-front aberrations compared with the younger group at their natural pupil diameters, owing to the effects of age-related miosis. These results suggest that although monochromatic ocular wave-front aberrations for a given pupil size increase with age, the reduction in CS with age is not due to this increase. PMID- 10474889 TI - Dynamical use of different sources of information in heading judgments from retinal flow. AB - The optic flow arising in the eyes of an observer during self-motion is influenced by the occurrence of eye movements. The determination of heading during eye movements may be based on the pattern of retinal image motion (the retinal flow) or on an additional use of an extraretinal eye-movement signal. Previous research has presented support for either of these hypotheses, depending on the movement geometry and the layout of the visual scene. A special situation in which all previous studies unequivocally have agreed that an extra-retinal signal is required occurs when the visual scene consists of a single frontoparallel plane. In this situation eye movements shift the center of expansion on the retina to a location that does not correspond to the direction of self-movement. Without extraretinal input, human observers confuse the center of expansion with their heading and show a systematical heading estimation error. We reexamined and further investigated this situation. We presented retinal flow stimuli on a large projection screen in the absence of extra-retinal input and varied stimulus size, presentation duration, and orientation of the plane. In contrast to previous studies we found that in the case of a perpendicular approach toward the plane, heading judgments can be accurate. Accurate judgments were observed when the field of view was large (90 degrees x 90 degrees) and the stimulus duration was short (< or = 0.5 s). For a small field of view or a prolonged stimulus presentation, a systematic and previously described error appeared that is related to the radial structure of the flow field and the location of the center of expansion. An oblique approach toward the plane results in an ambiguous flow field with two mathematically possible solutions for heading. In this situation, when the stimulus duration was short, subjects reported a perceived heading midway between these two solutions. For longer flow sequences, subjects again chose the center of expansion. Our results suggest a dynamical change in the analysis or interpretation of retinal flow during heading perception. PMID- 10474890 TI - Spectrally resolved white-light interferometry for measurement of ocular dispersion. AB - Spectrally resolved white-light interferometry was used to measure the wavelength dependence of refractive index (i.e., dispersion) for various ocular components. Verification of the technique's efficacy was substantiated by accurate measurement of the dispersive properties of water and fused silica, which have both been well-characterized in the past by single-wavelength measurement of the refractive index. The dispersion of bovine and rabbit aqueous and vitreous humors was measured from 400 to 1100 nm. In addition, the dispersion was measured from 400 to 700 nm for aqueous and vitreous humors extracted from goat and rhesus monkey eyes. An unsuccessful attempt was also made to use the technique for dispersion measurement of bovine cornea and lens. The principles of white-light interferometry, including image analysis, measurement accuracy, and limitations of the technique, are discussed. In addition, alternate techniques and previous measurements of ocular dispersion are reviewed. PMID- 10474891 TI - Mathematical modeling of retinal birefringence scanning. AB - Retinal birefringence scanning (RBS) is a new technique that is used to detect the fixation of the eye remotely and noninvasively. The method is based on analysis of polarization changes induced by the retina. In this study, the principles of RBS were mathematically modeled to facilitate a better understanding of the origins of the signals obtained. Stokes vector analysis and Mueller matrix multiplication were augmented with Poincare sphere representation. The cornea was modeled as a linear retarder. The foveal area was modeled as a radially symmetric birefringent medium. The model accurately predicted the frequency and phase of RBS signals obtained during central and paracentral fixation. The signal that indicates central fixation during RBS likely results from a combination of the radial birefringence of the Henle fibers and the overlying corneal birefringence. PMID- 10474892 TI - Spatial-scale contribution to the detection of mirror symmetry in fractal noise. AB - We investigated how the detection of mirror symmetry depends on the distribution of contrast energy across spatial scales. Stimuli consisted of vertically symmetric noise patterns with fractal power spectra defined by 1/f beta slopes ( 2 < or = beta < or = 5). While overall rms contrast remained fixed at 25%, symmetry-detection thresholds were obtained by corrupting the signal with variable amounts of noise with identical spectral characteristics. A first experiment measured thresholds as a function of spectral slope, and performance was found to be substantially facilitated in images with power spectra that characterize natural scenes (1.2 < or = beta < or = 3.2). In a second experiment, symmetry was removed from randomly chosen octave bands and replaced by noise with the same spectral profile. Results revealed that only in images with 1/f2 spectra does performance decrease by constant amounts across all frequency bands. Together, the results imply that symmetry mechanisms extract equal amounts of information from constant-octave frequency bands but lack the ability to whiten stimuli whose spectral slopes differ from those of natural scenes. Results are qualitatively well predicted by a multichannel model that (1) relies on spatial filters with equal-volume point-spread functions and constant-octave frequency bandwidths and (2) restricts the computation of symmetry to spatial regions whose dimensions are proportional to the filters' spatial scale. These findings are also consistent with the notion that mechanisms that mediate the perception of form exploit the ability of early vision to reduce second-order redundancy in natural scenes. PMID- 10474893 TI - Separating reflections from images by use of independent component analysis. AB - The image of an object can vary dramatically, depending on lighting, specularities, reflections, and shadows. It is often advantageous to separate these incidental variations from the intrinsic aspects of an image. We describe a method for photographing objects behind glass and digitally removing the reflections from the surface of the glass, leaving the image of the objects behind the glass intact. We describe the details of this method, which employs simple optical techniques and independent component analysis and show its efficacy with several examples. PMID- 10474894 TI - Independent-component analysis of skin color image. AB - The spatial distributions of melanin and hemoglobin in human skin are separated by independent-component analysis of a skin color image. The analysis is based on the skin color model with three assumptions: (1) Spatial variation of color in the skin is caused by two pigments, melanin and hemoglobin; (2) the quantities of the two pigments are mutually independent spatially; and (3) linearity holds among the quantities and the observed color signals in the optical density domain. The results of the separation agree well with physiological knowledge. The separated components are synthesized to simulate the various facial color images by changing the quantities of the two separated pigments. PMID- 10474895 TI - Theoretical development and experimental evaluation of imaging models for differential-interference-contrast microscopy. AB - Imaging models for differential-interference-contrast (DIC) microscopy are presented. Two- and three-dimensional models for DIC imaging under partially coherent illumination were derived and tested by using phantom specimens viewed with several conventional DIC microscopes and quasi-monochromatic light. DIC images recorded with a CCD camera were compared with model predictions that were generated by using theoretical point-spread functions, computer-generated phantoms, and estimated imaging parameters such as bias and shear. Results show quantitative and qualitative agreement between model and data for several imaging conditions. PMID- 10474896 TI - Pupil-size effects in fiber optic stellar interferometry. AB - The effects of finite telescope pupil sizes on the measurement of fringe visibility in fiber optic stellar interferometry are described. It is shown theoretically that the measured fringe visibility is equal to the cross correlation of the magnitude of the source's mutual coherence function with the cross correlation of the telescopes' effective pupil functions. If the telescopes' effective pupil diameters are not small compared with the width of the source's mutual coherence function, then the measured fringe visibility will be correspondingly distorted. The theoretical results are verified experimentally in a fiber optic Mach-Zehnder interferometer. PMID- 10474897 TI - Molecular phylogenetic evidence for a reversible morphogenetic switch controlling the gross morphology of two common genera of green seaweeds, Ulva and Enteromorpha. AB - Ulva and Enteromorpha are two of the most common, ubiquitous, and environmentally important genera of green seaweeds. They are widely regarded as easily distinguishable because of their dramatically different morphologies: Ulva species are flat, lettucelike blades two cell layers thick, and Enteromorpha species form hollow liquid- or gas-filled tubes one cell thick, which may also be highly branched. We present molecular phylogenetic analyses of nuclear ribosomal RNA ITS sequences from 39 samples representing 21 purported species within these two genera. The results clearly indicate that the two genera are not respectively monophyletic and that the characteristic Ulva and Enteromorpha morphologies have arisen independently several times throughout the evolutionary diversification of the group. The analyses demonstrate that this radical change in gross morphology can also happen within clades exhibiting sequence divergence typical of conspecific assemblages of this group. We suggest that this morphological flexibility is the result of some form of developmental switch that results in either blades or tubes, but that this putative switch must be activated relatively infrequently, since there is evidence that some lineages have retained their form for significant periods. This discovery suggests a possible new model system for study of the molecular mechanisms involved in the interplay between environmental stimuli and plant development. PMID- 10474898 TI - Molecular evolution of the CMT1A-REP region: a human- and chimpanzee-specific repeat. AB - The CMT1A-REP repeat consists of two copies of a 24-kb sequence on human chromosome 17p11.2-12 that flank a 1.5-Mb region containing a dosage-sensitive gene, peripheral nerve protein-22 (PMP22). Unequal meiotic crossover mediated by misalignment of proximal and distal copies of the CMT1A-REP in humans leads to a 1.5-Mb duplication or deletion associated with two common peripheral nerve diseases, Charcot-Marie-Tooth disease type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsies (HNPP). Previous molecular hybridization studies with CMT1A-REP sequences suggested that two copies of the repeat are also found in the chimpanzee, raising the possibility that this unique repeat arose during primate evolution. To further characterize the structure and evolutionary synthesis of the CMT1A-REP repeat, fluorescent in situ hybridization (FISH) analysis and heterologous PCR-based assays were carried out for a series of primates. Genomic DNA was analyzed with primers selected to differentially amplify the centromeric and telomeric ends of the human proximal and distal CMT1A REP elements and an associated mariner (MLE) sequence. All primate species examined (common chimpanzee, pygmy chimpanzee, gorilla, orangutan, gibbon, baboon, rhesus monkey, green monkey, owl monkey, and galago) tested positive for a copy of the distal element. In addition to humans, only the chimpanzee was found to have a copy of the proximal CMT1A-REP element. All but one primate species (galago) tested positive for the MLE located within the CMT1A-REP sequence. These observations confirm the hypothesis that the distal CMT1A-REP element is the ancestral sequence which was duplicated during primate evolution, provide support for a human-chimpanzee clade, and suggest that insertion of the MLE into the CMT1A-REP sequence occurred in the ancestor of anthropoid primates. PMID- 10474899 TI - Molecular phylogenetic analysis among bryophytes and tracheophytes based on combined data of plastid coded genes and the 18S rRNA gene. AB - The basal relationship of bryophytes and tracheophytes is problematic in land plant phylogeny. In addition to cladistic analyses of morphological data, molecular phylogenetic analyses of the nuclear small-subunit ribosomal RNA gene and the plastic gene rbcL have been performed, but no confident conclusions have been reached. Using the maximum-likelihood (ML) method, we analyzed 4,563 bp of aligned sequences from plastid protein-coding genes and 1,680 bp from the nuclear 18S rRNA gene. In the ML tree of deduced amino acid sequences of the plastid genes, hornworts were basal among the land plants, while mosses and liverworts each formed a clade and were sister to each other. Total-evidence evaluation of rRNA data and plastid protein-coding genes by TOTALML had an almost identical result. PMID- 10474900 TI - Patterns of molecular evolution among paralogous floral homeotic genes. AB - The plant MADS-box regulatory gene family includes several loci that control different aspects of inflorescence and floral development. Orthologs to the Arabidopsis thaliana MADS-box floral meristem genes APETALA1 and CAULIFLOWER and the floral organ identity genes APETALA3 and PISTILLATA were isolated from the congeneric species Arabidopsis lyrata. Analysis of these loci between these two Arabidopsis species, as well as three other more distantly related taxa, reveal contrasting dynamics of molecular evolution between these paralogous floral regulatory genes. Among the four loci, the CAL locus evolves at a significantly faster rate, which may be associated with the evolution of genetic redundancy between CAL and AP1. Moreover, there are significant differences in the distribution of replacement and synonymous substitutions between the functional gene domains of different floral homeotic loci. These results indicate that divergence in developmental function among paralogous members of regulatory gene families is accompanied by changes in rate and pattern of sequence evolution among loci. PMID- 10474901 TI - Genealogy of families of SINEs in cetaceans and artiodactyls: the presence of a huge superfamily of tRNA(Glu)-derived families of SINEs. AB - Several novel (sub)families of SINEs were isolated from the genomes of cetaceans and artiodactyls, and their sequences were determined. From comparisons of diagnostic nucleotides among the short interspersed repetitive elements (SINEs) in these (sub)families, we were able to draw the following conclusions. (1) After the divergence of the suborder Tylopoda (camels), the CHRS family of SINEs was newly created from tRNA(Glu) in a common ancestor of the lineages of the Suina (pigs and peccaries), Ruminantia (cows and deer), and Cetacea (whales and dolphins). (2) After divergence of the Suina lineage, the CHR-1 SINE and the CHR 2 SINE were generated successively in a common ancestor of ruminants, hippopotamuses, and cetaceans. (3) In the Ruminantia lineage, the Bov-tA SINE was generated by recombination between the CHR-2 SINE and Bov-A. (4) In the Suina lineage, the CHRS-S SINE was generated from the CHRS SINE. (5) In this latter lineage, the PRE-1 family of SINEs was created by insertion of part of the gene for tRNA(Arg) into the 5' region of the CHRS-S family. The distribution of a particular family of SINEs among species of artiodactyls and cetaceans confirmed the most recent conclusion for paraphyly of the order Artiodactyla. The present study also revealed that a newly created tRNA(Glu)-derived family of SINEs was subjected both to recombination with different units and to duplication of an internal sequence within a SINE unit to generate, during evolution, a huge superfamily of tRNA(Glu)-related families of SINEs that are now found in the genomes of artiodactyls and cetaceans. PMID- 10474902 TI - Molecular phylogeny and morphological homoplasy in fruitbats. AB - The present study evaluates the evolutionary framework of the Old World fruitbats based on the cytochrome b and 16S rRNA mitochondrial gene sequences from a wide range of taxa. Phylogenetic analyses indicated that morphology-based subfamilies and most suprageneric groups are nonnatural assemblages. They also support the existence of an endemic African clade of fruitbats. The discrepancy between the evolutionary relationships yielded by molecular and morphological data sets may be, at least in part, explained by the recurrent retention of primitive morphology (Rousettus-like) across different lineages. The maintenance of primitive characters in different groups of flying foxes, as well as morphological convergence in nectar-feeding bats and possibly also in short muzzle bats, may have led to high levels of homoplasy, resulting in misleading taxonomic arrangements. This may be particularly so with respect to high taxonomic levels based on morphological characters. PMID- 10474903 TI - Performance of the relative-rate test under nonstationary models of nucleotide substitution. AB - Relative-rate tests have previously been developed to compare the substitution rates of two sequences or two groups of sequences. These tests usually assume that the process of nucleotide substitution is stationary and the same for all lineages, i.e., uniform. In this study, we conducted simulations to assess the performance of the relative-rate tests when the molecular-clock (MC) hypothesis is true (i.e., there is no rate difference between lineages), but the stationarity and uniformity assumptions are violated. Kimura's and bias-corrected LogDet distances were used. We found that the computation of the variances and covariances of LogDet distances had to be modified, because the constraint that the sum of the frequencies of the 16 nucleotide pair types is equal to 1 must be imposed. Comparison of the rates of two single sequences (Wu and Li's test) or two groups of sequences (Li and Bousquet's test) gave similar results. When the sequences are long (> or = 500 nt), the test based on LogDet distances and their appropriate variances and covariances is appropriate even when the substitution process is not stationary and/or not uniform. That is, at the 5% significance level, the test rejects the MC hypothesis in about 5% of the simulation replicates. In contrast, if the sequences are short (< or = 200 bases) and highly divergent, the LogDet test is very conservative due to overestimation of the variances of the distances. When the uniformity assumption is violated, the relative-rate test based on Kimura's distances can be severely misleading because of differences in base composition between sequences. However, if the uniformity assumption held and so the base frequencies remained similar among sequences, the rate of rejection turned out to be close to 5%, especially with short sequences. Under such conditions, the test using Kimura's distances performs better than the LogDet test. The reason seems to be that these distances are less affected by a reduction in the number of sites than the LogDet distances because they depend on only two parameters. PMID- 10474904 TI - Patterns of genetic diversification within the Adh gene family in the grasses (Poaceae). AB - We investigated the evolutionary dynamics of the Adh gene family within the grasses (Poaceae), with the goal of using molecular evolutionary tools to understand the process of gene family diversification. We analyzed 21 Adh sequences representing a broad array of grasses. Phylogenetic analyses suggested that Adh duplicated into Adh1 and Adh2 before the radiation of the grasses roughly 65 MYA. Gene structure, including intron length, has varied little over this period. Conservation of intron length prompted investigation into the dynamics of intron evolution, particularly the ability of intron sequences to form secondary structures. Intron sequences did not have an extremely high or low minimum free energy of folding relative to permuted sequences, suggesting that individual Adh introns do not evolve under secondary structural constraints. For coding sequences, the diversification of Adh1 and Adh2 was marked by a shift in third-position G + C content. This shift may reflect differential selection for codon use. Diversification between Adh1 and Adh2 was also typified by a shift in nonsynonymous nucleotide substitution rates, but there was no evidence that relatively fast nonsynonymous nucleotide substitution rates in the Adh2 clade were a product of diversifying selection. Gene conversion may have played a role in retarding diversification of Adh1 and Adh2 in rice, but there is no evidence of gene conversion between paralogs in other taxa. Although the reasons for retention of two functional Adh genes remain obscure, we propose that a shift in gene expression was important for the retention of the two Adh gene copies within the grasses. PMID- 10474905 TI - Interspecific hybrid ancestry of a plant adaptive radiation: allopolyploidy of the Hawaiian silversword alliance (Asteraceae) inferred from floral homeotic gene duplications. AB - The polyploid Hawaiian silversword alliance (Asteraceae), a spectacular example of adaptive radiation in plants, was shown previously to have descended from North American tarweeds of the Madia/Raillardiopsis group, a primarily diploid assemblage. The origin of the polyploid condition in the silversword alliance was not resolved in earlier biosystematic, cytogenetic, and molecular studies, apart from the determination that polyploidy in modern species of Madia/Raillardiopsis arose independent of that of the Hawaiian group. We determined that two floral homeotic genes, ASAP3/TM6 and ASAP1, are found in duplicate copies within members of the Hawaiian silversword alliance and appear to have arisen as a result of interspecific hybridization between two North American tarweed species. Our molecular phylogenetic analyses of the ASAP3/TM6 loci suggest that the interspecific hybridization event in the ancestry of the Hawaiian silversword alliance involved members of lineages that include Raillardiopsis muirii (and perhaps Madia nutans) and Raillardiopsis scabrida. The ASAP1 analysis also indicates that the two species of Raillardiopsis are among the closest North American relatives of the Hawaiian silversword alliance. Previous biosystematic evidence demonstrates the potential for allopolyploid formation between members of the two North American tarweed lineages; a vigorous hybrid between R. muirii and R. scabrida has been produced that formed viable, mostly tetraporate (diploid) pollen, in keeping with observed meiotic failure. Various genetic consequences of allopolyploidy may help to explain the phenomenal evolutionary diversification of the silversword alliance. PMID- 10474906 TI - NMR diffusion and relaxation study of drug-protein interaction. AB - In this work, NMR diffusion and relaxation measurements are applied to the study of the interaction between the anti-inflammatory drug salicylate and the human serum albumin (HSA) in solutions. The self-diffusion coefficients and the spin lattice relaxation rates of salicylate are measured as a function of the concentration. The dissociation constant, Kd, for drug/HSA complexes and the number of binding sites, n, are evaluated. PMID- 10474907 TI - [41st annual meeting of the Japan Geriatrics Society. Kyoto, Japan. June 16-18, 1999. Abstracts] [In Process Citation] PMID- 10474908 TI - [The 42nd annual meeting of Japanese Society of Nephrology Yokohama, Japan. June 26-28, 1999. Abstracts] [In Process Citation] PMID- 10474909 TI - The core of competence. PMID- 10474910 TI - Audit of immunization policy and practice of health care workers within National Health Service Trusts in England and Wales. AB - The purpose of this study was to identify the variation in occupational health immunization policies and practice within NHS Trusts throughout England and Wales. Questionnaires were sent to 440 NHS Trusts and 279 were returned (a response rate of 63%). The results were compared with current Department of Health Guidelines. They highlighted the fact that NHS Trusts do not adopt a consistent approach to immunization practice and that these policies often do not reflect Department of Health Guidance. Of those responding, 249 (89%) stated that they would like additional guidance on immunization practice within the NHS workplace. The production of updated, evidence-based guidelines for immunization practice, may help to ensure that a more consistent approach is taken throughout the NHS. This would benefit both the Trusts and their employees who at present may be confused by being given different advice when moving between Trusts. PMID- 10474911 TI - Occupational asthma: a community based study. AB - The incidence and prevalence of occupational asthma has been extensively studied in industry settings and specialist clinics. However, it has been much less studied in the community. This study looked at the general practice notes of asthmatics in an attempt to assess the overall load of occupational asthma in the community. Eighty-six per cent of the patients with adult onset asthma in the practice population studied had at least one occupation recorded in their notes. Nearly a third of these (32%) were in jobs known to be significant causes of occupational asthma, yet a potential link between their occupation and symptoms had only been recorded in 18% of patients in these jobs. Overall 4% of the patients with adult onset asthma had been given a diagnosis of occupational asthma although in nearly half these cases the diagnosis had been made by a general practitioner and not a specialist. PMID- 10474912 TI - Ethical requirements for occupational health research--compliance arrangements for a single company in relation to a recent major nuclear industry study. AB - The media coverage given to occupational health studies in the field of ionizing radiation has, on occasion, been the cause of very real distress to radiation workers and their families. In response to this situation the Chief Medical Officers of the major UK nuclear companies developed an ethical policy for future involvement in research, based on the duty of care which researchers owe to a key customer of such studies: the worker. The policy consists of four principal elements: medical confidentiality; worker information; worker consent and the guarantee of the availability to the workers of pre-publication knowledge of the results. The policy issued in 1991/92 has achieved growing acceptance among researchers and medical journals, though the medical officers involved have been aware of some scepticism, particularly in relation to the practicalities of the dissemination of pre-publication information. The Record Linkage Study published in November 1997 marked a major piece of research work involving data from 120,000 radiation workers that had been carried out since the development of the policy. This paper reports on the successful compliance arrangements to meet the ethical requirements of that study within a single UK nuclear company, and is published to demonstrate that with commitment from researchers, the journal and occupational health staff such ethical requirements, and particularly the need for pre-publication information can be met in full. PMID- 10474913 TI - A review of the healthy worker effect in occupational epidemiology. AB - This review article aims to anatomize sources of the healthy worker effect (HWE) and to summarize advantages and limitations of several approaches frequently proposed to eliminate the HWE. Although the HWE is frequently addressed in the context of selection bias, our review suggests that the selection of occupational cohorts with advantageous health status would preferably be addressed as a source of confounding biases. The authors also conclude that the exclusion of unhealthy workers at employment and the study of active workers are the two main sources of HWE, and that the use of the general population as a comparison group in occupational epidemiology should be avoided if possible. The authors encourage investigators to make distinctions between the underlying factors related to the use of the general population as the comparison group in occupational epidemiology. PMID- 10474914 TI - Health and safety practices among farmers and other workers: a needs assessment. AB - The development of appropriate health and safety interventions for farmers and agri-workers is important world-wide but data on present practices and attitudes to change are lacking. A representative quota sample (n = 1,938) of the Irish population was surveyed on lifestyle practices and workplace risk assessment and control measures, in relation to chemical exposure, manual handling and machinery. Focus group discussions were conducted also with 47 representatives of national farming organizations. As compared with the general workforce, farmers had a significantly (p < 0.01) lower level of assessment of risk hazards associated with manual handling and machinery. Both farmers and employees in workplaces with less than 20 employees reported a significantly lower level of safety training. Male farmers had a particularly negative health profile with only 18% reporting regular dental checks, 26% practising skin protection and 29% taking regular exercise. Discussions indicated that barriers to change included low perceived susceptibility, lack of time and resources. Mental health issues were particularly highlighted. We conclude farmers differ significantly in many instances from the rest of the workforce in regard to occupational health and safety issues and specific interventions in key areas are required for the agri sector. PMID- 10474915 TI - Small bakeries--a cross-sectional study of respiratory symptoms, sensitization and dust exposure. AB - This cross-sectional study investigated the prevalence of respiratory symptoms and sensitization to dust components in 224 individuals in 18 small bakeries in Scotland. Each work practice in the bakeries was characterized by an assessment of dust exposure and assigned to a category with either a direct exposure to flour dust of an indirect exposure to flour dust. We found that work-related respiratory symptoms were significantly associated with specific IgE to wheat flour and amylase but not to exposure category (except for nasal/eye symptoms). However, specific IgE to wheat flour was significantly associated with exposure category. There was a higher prevalence of immunological sensitization, reporting of work-related respiratory symptoms and exposure to dust than in other studies and of the 144 personal dust sample results taken, 21 (14.6%) of the total exceeded 10 mg/m3, the substantial dust concentration as outlined by the COSHH Regulations. Follow-up of those with work-related asthma symptoms (questionnaire response) was inconclusive of the work-relatedness of their symptoms, although it did confirm respiratory morbidity. PMID- 10474916 TI - Exposure of health care workers to pentamidine isethionate. AB - Pentamidine isethionate is currently used for the prophylaxis and treatment of Pneumocystis carinii pneumonia. Its use has been associated with a number of symptoms in staff administering treatment, and there are some additional concerns about possible adverse health effects of long term exposure. The aim of this study was to quantify exposure of health care staff administering nebulized pentamidine to patients. Personal breathing zone and static air samples at the height of the patient's head were collected during the nebulization of pentamidine to nine sequential outpatients attending a haemophilia unit. These were analysed using a standard method allowing the exposure of staff to be estimated. The duration of treatment varied between 15 and 60 min. Personal breathing zone samples showed exposure to be between 2 and 100 micrograms/m3. Static samples showed the concentration of pentamidine in the room varied from 15 to 2,100 micrograms/m3. While these exposures were relatively low, they were higher than some other studies have reported, and may pose some risk of adverse effects to staff. Some simple measures could reduce staff exposure. PMID- 10474917 TI - Medical students and congenital colour vision deficiency: unnoticed problems and the case for screening. AB - The results are given of a questionnaire study to determine the range of difficulties that doctors notice in their work due to congenital colour vision deficiency (CCVD). The study is primarily qualitative. A questionnaire was sent to 40 self-selected doctors, 35 of whom were general practitioners (GPs). All were administered a number of colour vision tests to assess the type and severity of their deficiency. Many difficulties and some ways of overcoming them were reported. Those with a mild deficiency reported fewer difficulties and this relationship was significant. Twenty-three of the doctors also reported difficulties as medical students and their answers are given verbatim. The results are discussed in relation to other studies and data on colour vision. The reasons for and against screening medical students for this deficiency are considered and it is concluded that there is a strong case for screening. PMID- 10474918 TI - Communicating health risk. AB - Health risk communication is a two way interactive process that involves the exchange of information among interested parties about the nature, magnitude, significance, or control of a risk. Although it has only recently become a topic for scientific research, much has been learned in relation to the strategies and the techniques that contribute to effective health risk communication. In parallel, there has probably never been a time of greater need for effective training in health risk communication. The media and the general public are now very hazard conscious, subsequent to apparently regular events in the areas of public health, safety and environmental issues. Public concern regarding such issues is sometimes much less than experts feel to be appropriate, whilst at other times concern has outstripped the concern of the experts involved. Health professionals trained in the techniques of health risk communication are a vital resource in ensuring that the workforce or the population is properly informed so as to exercise appropriate decisions and actions in relation to hazard and risk. PMID- 10474919 TI - Do repetitive tasks give rise to musculoskeletal disorders? AB - Repetitive tasks can undoubtedly cause discomfort and pain, but whether they cause or worsen the pathology causing the pain is most uncertain. Research in this area is difficult as the 'work-related upper limb disorders' do not occur exclusively in workers and because there is no simple, reliable and reproducible test for most 'work-related upper limb disorders'. Furthermore many studies are difficult to interpret as they detect disease by the presence of symptoms: one would expect manual workers to complain of more symptoms than sedentary workers and symptom aggravation does not tell one anything about the causation of the underlying pathology. PMID- 10474920 TI - As others see us. AB - If it is to be fully effective, functional activity in business needs to see itself as an integral part of management and not as some specialist, separate adjunct. The so-called professions often find this difficult to contemplate, let alone achieve. In Unilever's UK operations, occupational health activity and staff are effectively integrated into general management and, as a direct consequence, the business benefits and the participants reach higher levels of job satisfaction. PMID- 10474921 TI - Auditing occupational medicine. AB - An important challenge facing the quality of practice in occupational medicine is a limited evidence-base, but equally important is the need to translate good evidence into high quality practice. Audit has an important role to play in addressing the determinants of variations in practice. Furthermore where the evidence is good enough to permit the development of valid practice guidelines, audit may help in improving education and standards of practice. External audit may have a role to play in ensuring conformity with service-level agreements and especially in addressing issues of quality which some management systems may fail to address. As more literature is published reviewing and achieving a consensus on the evidence-base for the practice of occupational medicine, and as more experience in audit is described, it can make an important contribution to quality in occupational medicine. PMID- 10474923 TI - Latex gloves. PMID- 10474922 TI - Contact dermatitis due to BCDMH in a hydrotherapy pool. PMID- 10474924 TI - Workers should not have their health damaged simply by attempting to earn a livelihood. PMID- 10474925 TI - [The state computerized information system for monitoring the epidemic process in Ukraine. The monitoring technology]. PMID- 10474926 TI - [The current methods for predicting the course of rheumatoid arthritis]. AB - One of the lines of inquiry into rheumatoid arthritis (RA) is a quest for markers capable of prognosticating the course of the illness. It is only genetic factors, the presence of rheumatoid factor, and activity of the illness that are most significant prognostic criteria. Showing much promise among the current approaches is DAS (Disease Activity Score) which relies on four variables (Richey index, number of swollen joints, ESR, and health index). The baseline and cumulative DAS values over a certain period of time can be used as prognostic factors. It is these criteria that the European League Against the Rheumatic Diseases Committee on Standardization of Clinical and Therapeutic Investigations have given approval to. They have been termed EULAR criteria. PMID- 10474927 TI - [Disorders in the functioning of the interferon system in rheumatoid arthritis]. PMID- 10474928 TI - [Baroreceptor regulation in chronic heart failure]. PMID- 10474929 TI - [Telethermography in pulmonology]. PMID- 10474930 TI - [The current concepts of the clinical picture, diagnosis and treatment of papillomavirus lesions of the female genitalia]. PMID- 10474931 TI - [The ecological epidemiological and immunological characteristics of infectious diseases in a large industrial region of the Don Basin (a review of the literature)]. PMID- 10474932 TI - [The effect of ecological and microbiological factors on the health status of pregnant women with pyelonephritis and on their newborn infants]. AB - An analysis was performed of history, course of pregnancy, parturition, and condition of the newborn babies in female patients with pyelonephritis depending on the microbiological factor and environmental situation in the zone of residence. The pregnant women living under adverse environmental conditions display high levels of endocrine, cardiovascular, gastrointestinal disorders, and of tonsillitis. In the majority of cases, pregnancy is noted to be complicated by anemia (76.7%) and fetoplacental incompetence (62.9%), with infants being born in asphyxia presenting with signs of hypotrophy, congenital infection. Of the above infants, 37% develop postnatal inflammatory conditions. Two variants were shown to play a part in the etiology of pyelonephritis: monoetiological one marked by predominance of Staphylococcus aureus and Escherichia coli and polyetiological variant characterized by predominance of Candida fungi, Staphylococcus aureus, and mycoplasma. Irrespective of the microbiological factor, the female patients with pyelonephritis demonstrated high levels of premature birth, had a history of infertility, and were presenting with genital and extragenital pathologies. PMID- 10474933 TI - [The functional status of the thyroid system in those irradiated as a consequence of the accident at the Chernobyl Atomic Electric Power Station (based on screening study data)]. AB - A screening study was made of functional indices for the status of the thyroid system in 1097 persons who took part in the elimination of the effects of the Chernobyl accident (PChAEE) during the "iodine" and "non-iodine" periods of irradiation. In 94.6% of the "iodine" period PChAEE, the basal levels of the thyroid-stimulating hormone (TSH) were in the normal range, 4.79% displayed a rise in the parameter. 90.06% of the "non-iodine" period had normal range TSH, 8.77% exhibited augmented levels. 94.68% of the "iodine" period PChAEE showed normal range for the basal levels of free thyroxin, in 3.72% there was a rise, in 1.6% a decrease in the values. Among the examinees of the "non-iodine" period, the levels of free thyroxin were within normal range in 92%, above norm in 5%, decreased in 3%. The level of antibodies to thyroglobulin in 86.38% of the "iodine" period examinees was in the normal range, in 90.99% of the "non-iodine" period it was also normal. There were no statistically significant differences suggesting effects of radiation factors of the "iodine" period of irradiation on the functional indices for the hypophysis-thyroid system during the first 7 years following irradiation. PMID- 10474934 TI - [The immunological reactivity of children living under conditions of the action of low doses of ionizing radiation and of industrial waste]. AB - We undertook the present studies to gain insights into the immunological reactivity of children exposed to small doses of ionizing radiation and salts of heavy metals. A total of 1500 children were examined who live in Sumy Province areas affected by varying levels of radioactive contamination. Increased morbidity, with thyroid hyperplasia being noted more often than not, has come to be recorded among children residing in the territories contaminated with discharges of salts of heavy metals and small doses of ionizing radiation. Imbalance of the thyroid and immune systems has been disclosed. The degree of immunological disorders was found to be related to specificities of activity of the hypophysis-and-thyroid system. Those children who are permanent residents in health-hazard localities exhibit a significant rise in blood level of interferones. Doing away with thyroid insufficiency will, we believe, help in returning the immune disorders in children to normal. PMID- 10474935 TI - [The role of heredity in the prevalence of arterial hypertension and its risk factors]. AB - Studied in the rural population both men and women residing in the territories of Chernigov Province was prevalence of aggravated heredity in respect of myocardial infarction, cerebral insult, arterial hypertension, diabetes mellitus, and sudden non-murderous death. More frequently encountered in the above population is aggravated heredity in respect to arterial hypertension, with significant predominance being recordable of maternal-lineage heredity. The prevalence of arterial hypertension in probands with hereditary aggravation is much greater than in the population, especially so among women. Positive correlation was disclosed between arterial hypertension with excess consumption of table salt and overweight in probands with hereditary aggravation. PMID- 10474936 TI - [The fatty acid composition of the venous and arterial blood lipids in the complications in the acute period of myocardial infarct]. AB - Lipid fatty-acid composition was determined as was the level of free cholesterol in serum and red cells of venous and arterial blood in patients with myocardial infarction presenting with certain complications. The most significant changes in fatty-acid composition of blood serum lipids were in pulmonary edema and pneumonia, with those in the cell lipid complex being recordable in cardiogenic shock and ventricular fibrillation. Fatty-acid composition of the arterial blood lipids is more balanced compared to the venous blood in pulmonary edema, cardiogenic shock, ventricular fibrillation; the opposite tendency is observed during the development of pneumonia. PMID- 10474937 TI - [The evolution of the clinical course of infectious endocarditis]. AB - As many as 188 records of IE patients were studied as were 67 autopsies of deceased persons. Significant changes in the clinical course of IE were disclosed over the past 10-15 years versus previous 15: more prominent role of staphylococcal and gram-negative flora in the etiology of the illness, with microbial associations-caused IE cases having become more common. Diagnostic value of hemoculture did not appear to show much promise. The following items were found to be important risk factors for IE: prior viral and bacterial infection, invasive interventions, and chronic somatic diseases. There prevailed those IE forms having appeared on the intact heart valves in elderly persons and those at a mature age. In the course of the illness there tended to be more toxic and allergic manifestations, thromboembolic complications, and extensive vascular pathology. PMID- 10474938 TI - [The hemodynamic status of patients with ischemic heart disease under long-term treatment with calcium antagonists]. PMID- 10474939 TI - [The characteristics of the sympathico-adrenal and kinin systems in engineers in the early stages of arterial hypertension]. AB - Overall seventy-two engine-drivers with borderline arterial hypertension (BAH) and stage I arterial hypertension (AH) were studied for the condition of the sympathoadrenal and kinin systems before and after trip. Both groups exhibited a rise in the activity of the sympathoadrenal system before trip, with still greater rise being recordable after trip. BAH and stage I AH subjects demonstrated an increased activity of the kinin system of the kidneys before trip and decreased one after trip. Suppression of the kinin system of the kidneys with simultaneous progression of pressor effects leads to decline in adaptation to physical and psychoemotional overloads in those engine-drivers presenting with early stages of AH. PMID- 10474940 TI - [The effect of local vibration on central and cerebral hemodynamics]. AB - Effects of vibronoise industrial factor on central and peripheral hemodynamics were studied as was the possibility of correction of changes by antagonists in short-action calcium channels. It is shown that 68.9% of those workers under chronic exposure to industrial vibration and noise have hyperkinetic-type blood circulation, 21.3% eukinetic, 9.8% hypokinetic type. Besides, workers exhibit diminution in general cerebral bloodflow presenting with spastic states of great and small arterial vessels. There is noted a length of service-related increase in percentage of persons presenting with eu- and hypokinetic types of circulation and advancing diminution in general cerebral bloodflow with progression of spasm of arterial vessels being recordable. Antagonists of short-action calcium channels have a beneficial effect on central and cerebral hemodynamics. PMID- 10474941 TI - [The characteristics of erythropoiesis in extremely premature neonates in the 1st 2 months of life]. AB - Based on studies of morphometric indices for red blood and morphometric characteristics of erythrocytes, an evaluation was carried out of the condition of erythropoiesis in 109 extremely premature newborn babies (gestational age 26 to 32 weeks, body mass at birth 800-1600 gr). Noted in the above babies were markedly decreased values for the red blood during the first 1 to 1.5 mo of life and apparent relatedness of the rate of decrement to gestational age and concurrent pathology (infections, intraventricular hemorrhage, grave breathing disorders, hypotrophy). Besides, a low reticulocytic response was recordable to the developing anemia and high degree heterogeneity of the cellular (erythroid) population. The data secured suggest failure of erythropoiesis in extremely premature neonates, the reason for which phenomenon might be too little production of erythropoietins, which fact prompts us to look at the pathogenesis of early anemia and potentialities of correcting thereof from a new angle. PMID- 10474942 TI - [The clinico-metabolic characteristics of the course of a chlamydial infection in newborns]. AB - A clinical and laboratory investigation was done in 68 full-term newborn babies, with clinical forms of Chlamydia infection having been identified. There have also been studied cellular and humoral links of immunity, hormonal profile, system of peroxidation, antioxidant defence. In Chlamydia infection in full-term newborns presenting with acute bronchopulmonary pathologies during the acute phase of the illness, signs are noted of activation of processes of free-radical lipid oxidation against the background of retardation of the antioxidant defense, with synthesis of hormones of the thyroidal, hypophyseal and adrenal systems being on the increase. Metabolic changes are directly related to immunologic responsiveness which is characterized by weakness of primary immune response. The degree the above changes make themselves known depend on the clinical form of Chlamydia-associated infection. The identified metabolic changes may be the basis of causative factors for change in capacity of newborn infants with chlamydial infection to adapt. PMID- 10474943 TI - [The development of a coagglutination reaction using staphylococcal peptidoglycan for the diagnosis of staphylococcal infection]. AB - Our objective in this study was to develop and assess the diagnostic value of a coagglutination test with making use of peptidoglycane of Staphylococcus aureus in the identification of diseases of staphylococcal etiology. A total of 166 patients with diseases of staphylococcal etiology were examined. A test was elaborated of coagglutination with making use of peptidoglycane of Staphylococcus aureus for a differential identification of antibodies to peptidoglycan in healthy persons and patients with staphylococcosis. PMID- 10474945 TI - [Tesigraphic studies of the blood serum in psoriasis patients]. AB - 82 patients with psoriasis and 10 apparently healthy subjects (group of control) were examined. After analysis of the psoriasis patients blood serum thesigrammes in this study 3 distinct types were identified: tree-like one (49%), bush-bunch type (44%), and polymorphous type (7%). In a microscopic investigation, thesigrammes of psoriasis patients differed from those of healthy subjects in respect of morphostructural features in spite of macroscopic similarity. While studying thesigrammes during the time-related course of the illness significant differences depending on the stage of psoriasis were disclosed. The results secured permit asserting that the thesigraphic method of investigation into the blood serum can be used for the diagnosis of the condition, objectivization of control over the effectiveness of treatment and prognostication of the course of psoriasis. PMID- 10474944 TI - [The correction of the immunohormonal disorders in patients with hyperplastic processes in the endometrium]. AB - Effects were studied of extremely high-frequency electromagnetic radiation (EHF ER) on indices for the immune and endocrine systems in 48 patients with hyperplastic processes in the endometrium. It is ascertain that along with the established normalization of parameters characterizing the above systems there occurs a return of correlations back to normal, which fact permits using EHF ER as an immunocorrective factor. PMID- 10474946 TI - [The reinnervation of skeletal muscles]. AB - Experiments were conducted to study different methods of reinnervation of predenervated muscle, for which purpose three groups of animals were used. The first group (reinnervation as to Brunelli) served as a kind of control. In the second group, side notches were cut in the nerve-neurotizator before inserting it into the muscle, in the third group, endomysium and perimysium were defibered at the site of insertion into the muscle of the nerve-neurotizator. The obtained results showed that in the second and third groups processes of reinnervation proceed faster by far and in a more efficient way. This is because the nerve neurotizator has side cuttings in it, which fact promotes the ingrowth into the muscle tissue of a great many of nerve conductors, and because the skeletal muscle neurotropism gets enhanced at the expense of the connective tissue being formed in sites of defiberation of endomysium and perimysium. The secured results permit recommending the proposed ways and means for reinnervation of denervated muscles in a clinical setting as more efficacious than that by Brunelli. PMID- 10474948 TI - [The diagnosis and treatment of acute pancreatitis in patients with atherosclerosis of the vessels]. AB - As many as 988 patients with acute pancreatitis were given hospital treatment at the clinic of general surgery, its total duration being three years. Atherosclerosis of vessels was identified in 616 (62%). Difficulties with clinical diagnosis occurred in patients with moderately severe to severe degree of atherosclerosis (62%). Laparoscopy appeared to be undoubtedly superior to other diagnostic aids employed. Conservative therapy was found to be effective in 557 patients. Postoperative case mortality was 6 patients with destructive forms of pancreatitis. PMID- 10474947 TI - [The quantum gravitational therapy of myocarditis]. AB - Complex therapy of myocarditis of rheumatic and non-rheumatic genesis using intravascular laser irradiation of blood, quercitrol, and enterosgel has an antiinflammatory, antioxidant action, improves myocardial contractility, is endowed with an antiaggregatory activity. The above therapeutic complex permits the reduction of the non-steroid antiinflammatory drugs intake as well as of the average time of hospital treatment by 2 to 3 days, it also makes for an earlier medical and social rehabilitation of patients. PMID- 10474949 TI - [The optimization of treatment outcomes in patients with the Mallory-Weiss syndrome]. AB - Examined in the study were results of conservative and surgical treatment of 202 patients presenting with Mallory-Weiss syndrome. Etiological factors of the syndrome were studied, investigations designed to study the regional bloodflow of the stomach in the zone of ruptures and intact areas, the motor and acid producing functions of the stomach were conducted. Based on the conducted general clinical and special examination of patients with Mallory-Weiss syndrome the importance was proved of ischemic atrophic gastritis, discoordination of the motor and evacuatory function of the esophagus and duodenum in the genesis of the above syndrome, which disorders tend to be more manifest in sudden impacts of predisposing factors. In this connection, an etiopathogenetic approach has been developed towards treating patients such as mentioned above. Surgical treatment was being undertaken only when specifically indicated. PMID- 10474950 TI - [Dynamic radiation telethermometry in the diagnosis of acute appendicitis]. AB - Density was studied of heat flow from the area of the anterior abdominal wall in 167 patients with acute appendicitis. A novel methodological approach has been developed to the diagnosis of acute appendicitis making use of remote thermometry with the unit "Radiation heat flows meter". Recording of heat flows from the region of the anterior abdominal wall helps in the diagnosis of acute appendicitis, allows some preliminary judgement about the degree of affection of the organ, permits choosing relevant policy of managing a particular patient. The method can be employed in planned and urgent surgery. PMID- 10474951 TI - [An analysis of the information value of certain factors in the prognosis of the severe form of bronchial asthma in adolescents]. AB - A total of 150 children of pubertal age, who had bronchial asthma were examined. An in-depth analysis of those factors promoting the development of a grave form of the condition in the examined children permitted the selection of five of them, that have statistically significant effects on formation of this pathology: sex, concurrent skin afflictions, early onset of the illness. Improper treatment, abuse of antibiotics. With the aid of the logistic regression technique there has been developed a mathematical model that permits the prognostication of severity of bronchial asthma course at pubertal age to be carried out on an individual basis. PMID- 10474952 TI - [The prevention and treatment of a Candida-mycosis type of dysbacteriosis in chronic bronchitis in older patients]. AB - Changes were studied in the clinical picture and microecological setting of the large intestine in elderly and old patients with chronic bronchitis (n = 17) against the background of antibacterial therapy that leads to the development of dysbacteriosis, including that with increased content of Candida. Shown in elderly and old patients with chronic bronchitis is efficacy of the outlined treatment option for the prophylaxis of candidiasis type dysbacteriosis (diphlucan, linaex, decamevite, nitroxoline). PMID- 10474953 TI - [The local immunity of the urinary system in patients with recurrent urolithiasis]. AB - 51 patients with urolithiasis complicated by pyelonephritis in the active phase of inflammation were studied for the condition of local immunity by determining the urine content of secretory immunoglobulin A (SIgA) under conditions of combined treatment with making use of phlogenzyme, a drug of II-generation systemic enzymotherapy (SE). Recordable in this patients population was a marked increase in the urine level of SIgA. Incorporation in a combined treatment of phlogenzyme results in normalizing the status of the urinary system local immunity. Evidence has been obtained on the lack of parallelism in the dynamics between the serum IgA content and urine concentration of SIgA, which fact suggests independence of local immunity. Our theory is that an appreciable increase in the urine level of SIgA in patients with urolithiasis concurrent with pyelonephritis may have an important part to play in the genesis of nephrolithiasis. PMID- 10474954 TI - [The characteristics of the pathogenesis and clinical forms of the course of glomerulonephritis in adolescent patients]. AB - Overall, 138 patients at teen age and youths were studied for the clinical course of their glomerulonephritis. In adolescence, the onset of glomerulonephritis presents as urinary syndrome with hematuria and nephrotic syndrome. Acute glomerulonephritis with hematuria is accompanied by a low level of proteinuria and slow progression. Apparent benefit from the treatment occurred in 73 +/- 5% of patients (complete and partial clinical and laboratory remission), unsatisfactory results of treatment were recorded in 27 +/- 5% of patients. Persistence of nephrotic syndrome in patients demonstrating unsatisfactory results of treatment coincide with far greater frequency of unfavourable morphological forms of glomerulonephritis (mesangiocapillary and focal-segmentary glomerulosclerosis (hyalinosis), changes in canaliculi and interstitium, with loss of the functional reserves of the kidneys and effect of plasmapheresis on hyperlipidemia being recordable. PMID- 10474955 TI - [The immunological aspects of a hip joint lesion in childhood]. AB - The paper presents an in-depth analysis of immunological mechanisms of the hip joint synovitis development in children. Studied in the pediatric patients with transient coxitis, Perthes disease, and tuberculous coxitis (n = 54, 15, and 15 respectively) with the aid of flow cytofluorometer FACStar PLUS was the subpopulation of immunocompetent cells with a wide spectrum of MCAB. Controls (n = 35) were sex and age-matched. Comparative analysis of immunological parameters in the above children disclosed a toxic-and-allergic origination of transient synovitis with predominant affection of the T-cell link of immunity (T-helpers failure and activation of T-suppressors). Joint mucose injury was noted to be developing against the background of reduction of serum IgA. Activation of non specific mechanisms (natural killers, increase in SDH activity) is of compensatory character. Mechanisms of activation of cell processes (expression of panmitogenic receptor CD4+ antigen) in Perthes disease warrant further studies. In patients with tuberculous coxitis, immunological incompetence develops in the presence of cytotoxic reactions and activation of the B-link, with the production of IgM being on the increase but with no essential changes in the IgG fraction. It is necessary that selective stimulators of T-helpers be included into the treatment scheme together with drugs capable of exerting a selective effect on the T-link of immunity. PMID- 10474956 TI - [New approaches in the treatment of speech disorders in children with an organic brain lesion]. AB - Speech disorders encountered in children presenting with neurological pathology make their medical and social rehabilitation a real challenge. Overall sixty children were examined. Of these, 38 had infantile cerebral paralysis, 22 that being presented with sequelae of the hypoxic affection of the brain in ante- and perinatal period, who exhibited speech disorders (alalia, dysarthria, retarded speech development). Described in the paper are features of bloodflow disorders in intracranial and vertebral arteries as evidenced by ultrasonic Doppler technique and changes in brain bioelectrical activity according to findings from electroencephalomapping. In 53 patients, a positive effect was obtained as a result of therapy conducted in accordance with the stimulation type techniques (electroscalp therapy against the background of intramuscular administration of nicotinic acid plus laser therapy, transcutaneous electrostimulation, employment of such drug preparations as cogitum, nero-force, sirdalud). PMID- 10474957 TI - [The use of sorption therapy in the combined treatment of exudative erythema multiforme]. AB - Polymethylsiloxan, a sorbent, was used in combination with enterosorption with thermal alkaline mineral water in a complex treatment of patients (n = 37) with multiform exudative erythema. During the 45-min therapeutic session the sorbent applicator was changed three times. The above therapy makes for rapid healing of erosions, reduces treatment time periods, prevents recurrences irrespective of the causative factors of inductors of the disease under study. PMID- 10474958 TI - [Serum sickness in diphtheria]. AB - As many as 2247 patients with different clinical forms of diphtheria were examined. Antidiphtheric serum (ADS) was administered in 1556 children, the dosage being determined by condition of the patient. Serum sickness developed at day 7 to 9 in 24 (1.5%); 10 patients were found to run a mild course, 14- moderately severe. 6 patients had allergic reactions: 3--to antibiotic (penicillin), urticaria type, 1--to pertussoid-tetanic anatoxin, 2 had pollinosis type reaction. Thus, serum sickness has practical value, which fact requires a detailed allergic history together with skin tests to be performed before the administration of ADS. PMID- 10474959 TI - [Diagnostic problems in diphtheria infection]. AB - The problem of diagnosis of diphtherial infection has come to acquire especially significance at the present stage because here in Ukraine we have no legitimate standard procedure for identification of a case of diphtheria and because interpretation of results of bacteriological and serological studies in patients who might have diphtheria is often faulty. The diagnosis of atypical cases of mild localized diphtheria in the vaccinated subjects relies on the presence of chronic ENT-affections and a history of predisposition to sore throat. It is necessary that parallel culturing be done of smears obtained from the nasopharynx for diphtheria and pathogen microflora in order that no possibility of angina of other microbe etiology might be entertained. Isolation of nontoxicogenic strains C diphtheria in the absence of clinical signs of diphtheria should not prompt us to diagnose a patients as having this problem. Serologic investigation of the patients serum in the passive hemagglutination test for the presence of antitoxic antibodies can be recommended for use to verify the vaccination history in those having been taken ill rather than to aid in the differential diagnosis of diphtheria and bacteria-carrier states. Results of bacteriological and serological investigations do not give much ground for rejection or confirmation of the diagnosis made. PMID- 10474960 TI - [The course of pregnancy and labor in women with combined anemia of pregnancy and late gestosis]. AB - The course of pregnancy in anemia of pregnancy current with late gestosis is characterized by an early development of fetoplacental insufficiency, a greater risk for hypertrophy of the fetus, hemorrhage during the third and early postpartum period, septic puerperal complications. Early correction of the above pathological states can diminish the number of disorders induced by them in the organism of the mother and fetus alike. PMID- 10474961 TI - [The diagnosis and treatment of background and precancerous diseases of the vulva]. AB - Methodological approaches have been developed toward raising the informative value of cytological diagnosis. There have been identified pathogenetic variants of background and precancerous affections of the vulva, their leading factors being viral infection, function disorders of the hypothalamic centres, hypoestrogenemia, hepatic and thyroidal abnormalities. The effectiveness of the treatment scheme developed with taking account of the above-stated items is 97.3%. PMID- 10474962 TI - [The use of enterosorption and plasmapheresis during the chemotherapy of lung cancer patients]. AB - As many as 44 patients with stage IIIB and IV carcinoma of the lung (n = 27 and 17 respectively) were studied for clinical and laboratory manifestations of endogenous intoxication syndrome during the administration of CAPF chemotherapy treatments. The use for detoxicating purposes of siliceous enterosorbents and plasmapheresis both alone and combined resulted in alleviation of gastrointestinal and hematological complications and permitted improving the quality of the patients' lives. PMID- 10474963 TI - [Kaopectate in the combined treatment of patients with intestinal infections]. AB - The paper presents results of multiple-modality treatment of patients with acute intestinal infections with making use of the drug preparation Kaopectate incorporating activated attapulgite which is a natural mixture of hydrates of magnesium silicate and silicate of alumina. The above drug has been shown to be endowed with a antidiarrheal and detoxifying activities. PMID- 10474964 TI - [The use of Flamikar in patients with chronic hepatitis of alcoholic etiology]. AB - Clinical efficacy was studied of flamicar in patients with chronic hepatitis of alcohol etiology. Flamicar by 21-day courses was found to lower the degree of clinical symptoms of the illness, making for improvement of somatic condition of the patient, returning the indices for the functional state of the liver to normal. Thus, flamicar is an effective remedy for patients with chronic alcohol hepatitis. PMID- 10474965 TI - [The acute course of generalized sarcoidosis]. PMID- 10474966 TI - [The current views on the role and importance of phytotherapy in the structure of scientific medicine]. AB - Phytotherapy, in spite of a long-continued employment in empirical and scientific medicine, does not have distinct scientific doctrine, conceptual strategies and tactics of use in practical treatment. Ways and means of phytotherapy are more often than not regarded as adjuncts to pharmacotherapy. In connection with formation during recent decades of new concepts of pathogenetic essence of pathological processes as of combined disturbances in biochemical reactions,--at the membrane and cell level in the first place, the role and value of phytotherapy as an independent method and means of metabolic therapy is considered in a new perspective. PMID- 10474967 TI - [The use of reflexotherapy in the initial forms of cerebrovascular insufficiency in persons who suffered as a consequence of the accident at the Chernobyl Atomic Electric Power Station]. AB - A comprehensive evaluation has been done in 120 patients presenting with primary forms of cerebrovascular insufficiency (PFCVI), who had become victims of Chernobyl accident. PFCVI diagnosis was made on the basis of findings from the clinical investigation, rheoencephalography, electroencephalography, echoencephalography, iridodiagnosis, electropuncture diagnosis as recommended by Nakatani, variation pulsometry, study into the vegetative nervous system. Functional inadequacies were disclosed in the bodily regulatory-adaptive system, specifically, the system of regulation of cerebral circulation. Cerebral symptoms in PFCVI are a reflection, first of all, of the brain nonspecific systems dysfunction. All this was taken into account in working out schemes of treatment for the above-named category of patients, incorporating lasero- and acupuncture. Exposure to the treatments described resulted in a noticeable improvement in 78% of the patients, and improvement in the rest of the series. High efficiency of the proposed approach to the treatment of PFCVI patients having become victims of Chernobyl accident suggests its pathogenetic significance. PMID- 10474968 TI - [Experience in the use of a method of intracutaneous oxygen and zonal reflexotherapy in the practical treatment and rehabilitation of persons who suffered as a consequence of the accident at the Chernobyl Atomic Electric Power Station]. PMID- 10474969 TI - [The effect of reflexotherapy on the hemostatic system of patients with glomerulonephritis]. AB - A total of 104 patients with different forms, variants and stages of glomerulonephritis were both examined and treated. Acupuncture has a normalizing effect on the system of hemostasis which was manifested by a decrease in the level of hyperfibrinemia, thrombinemia, activation of fibrinolysis and impairment of aggregation properties of blood plates. PMID- 10474970 TI - [The characteristics of the work activities, their autonomic support and the work capacity of coal mine workers ill with chronic bronchitis]. AB - In miners with chronic bronchitis, diminution of bodily energy reserves does not permit optimizing their functional state in full measure. Patients are provided with ample opportunity to realize various forms of behaviour in order that they might continue their labour activities under usual conditions they are accustomed to. Exclusion from the activity those labour operations of high energy content, lowering of density of work, and increase in duration/number of breaks for rest- all such measures serve to secure professional activity. From the standpoint of self-preservation of the organism the above-named changes can be regarded as quite expedient since they minimize the action of adverse factors of the labour process. Nevertheless, this does not save the miners weakened by their illnesses from development of unfavourable states of overstrain during the process of their work. PMID- 10474971 TI - [The basis of the terms "therapeutic-prophylactic and medical institutions"]. PMID- 10474972 TI - [The scientific and methodological aspects of creating and introducing uniform standards for medical technologies]. PMID- 10474973 TI - [The 70th anniversary of the founding of the Department of the Propedeutics of Internal Diseases of the National Medical University]. PMID- 10474974 TI - Set-up error & organ motion uncertainty: a review. AB - Conformal radiotherapy allows improvement in the treatment outcome due to increased targeting accuracy through advanced beam shaping techniques to precisely conform radiation dose to the geometry of the tumour. Treatment set-up and organ motion uncertainties are unavoidable factors that are limiting increases in accuracy and have to be accounted for in conformal treatment planning. The magnitudes of set-up errors and organ motion uncertainties for specific sites, and using various set-up techniques, have been quantified in the literature. However, the parameters used with these measurements and the presentation of the data has differed between studies for the same site. The purpose of this paper is to analyse and combine the published material into a uniform format and to display typical reported values of set-up and organ motion uncertainties. Values measured under similar conditions were averaged across studies. The results of this analysis illustrate (1) variability in the parameters used for measurements across studies, (2) typical motion ranges of the prostate, kidneys, liver and diaphragm, (3) typical means and standard deviations for set-up errors associated with the prostate, pelvis, brain, head and neck, thorax, rectum and breast and (4) a brief review of the common methods to lower or account for these uncertainties. PMID- 10474975 TI - A method to predict the effect of organ motion and set-up variations on treatment plans. AB - Dose distributions calculated by commercial treatment planning systems do not allow incorporation of the effects of patient position variation or organ motion throughout the course of radiation therapy treatment. We have established a convolution-based method, which enables us to display dose distributions using a commercial treatment planning system that can take into account target movement. An example of the method applied to a prostate treatment plan is presented. For the method to be of clinical use it requires assessment of the parameters leading to target movement in a scientific manner in the same treatment department that it is to be used. It is not sufficient to rely on published data especially that relating to set-up accuracy as this has been shown to vary widely from centre to centre. We believe that with appropriate movement data, a convolution-based approach can lead to more optimal radiation margins around clinical target volumes (CTV). Optimal margins will help prevent geometric misses as well as ensure that the amount of critical late reacting normal tissues surrounding the CTV irradiated is minimised. Optimal margins cannot be guaranteed with the more conventionally used "rule of thumb" techniques for placing a planning target volume around the CTV. PMID- 10474976 TI - An investigation into the impact of anatomical variation upon mean glandular dose produced within a standard breast. AB - By using the EGS4 Monte Carlo computer program the normalised mean glandular dose (MGD) was calculated for a breast model that is intended to reflect the composition of an "average" breast. The reliability of the calculation was established by comparing the predictions with previously published values for a breast model. The breast model used was then altered in order to reflect the possible extremes in glandular distribution that could occur within a compressed breast. These results show that the MGD could vary by up to a factor of four depending upon where the majority of glandular tissue is located within the breast. The impact of this variation in MGD upon risk assessment within mammography is then discussed. PMID- 10474977 TI - A survey of radiation doses to adults from diagnostic radiology in Victoria. AB - This study examines doses to adult patients from diagnostic radiology. Measurements were made at 37 hospitals and private radiology centres in the state of Victoria. Skin entrance doses in air were measured for the exposure factors used by the radiographer for an average size patient for the following procedures: abdomen AP; pelvis AP; lumbar spine AP, lat, LSJ; and skull AP, lat. There was a large range of doses observed for each particular procedure. Factors contributing to the range of doses were identified. Guidance levels for adult radiography based on the third quartile values of the skin entrance dose have been adopted for use in Victoria. PMID- 10474978 TI - Real-time rotation vectors. AB - Rotation vectors are a useful way of describing eye position without reference to arbitrary axes of rotation since any eye position can be reached from the reference position by rotation about a single axis. A real-time display of rotation vectors would not only help to acquire more reliable data, but would also widen the range of possible eye movement experiments. We describe a novel PC based data acquisition and analysis system which calculates and displays rotation vectors, velocity vectors and Listing's plane in real-time using voltages obtained from a two field coil system. The system was implemented using LabVIEW and optimised using Code Interface Nodes. Off-line processing can be sped up by varying parameters that indicate the amount of available RAM. During processing Listing's plane data can be rotated horizontally, vertically and torsionally. A computer controlled laser target changes position randomly every half second and so the targets are evenly spread, producing an appropriate range of eye positions which are used to calculate Listing's plane. PMID- 10474979 TI - ECG of the month. Turned about. Dextrocardia. PMID- 10474980 TI - ENT issues in pregnancy. AB - Pregnancy initiates a unique set of physiologic changes in a woman's body. These changes are commonly manifested as complaints concerning the head and neck. From common findings such as rhinitis, epistaxis, and cutaneous lesions to more rare disorders of the inner ear, physicians should be familiar with these conditions for optimal reassurance, expectant management, or treatment of the gravid female. It is especially important to be aware of the possible effects of any medication, including over-the-counter remedies, on the mother and fetus and to consult the treating obstetrician prior to writing any prescriptions. Many of these disorders will be discussed with an emphasis on rhinitis during pregnancy to illustrate the complexity and controversy surrounding many of the conditions. Finally, there will be a directed focus on commonly used pharmaceuticals in otolaryngology and their safety during pregnancy. PMID- 10474981 TI - The journal 100 years ago. July 1849 and 1899. PMID- 10474982 TI - Characteristics of women in Louisiana who give birth without receiving prenatal care. AB - The percentages of Louisiana female residents who did not receive any prenatal care before giving birth in 1985, 1990, 1992, and 1995 are derived from birth certificate data. The amount of change between the percentages of women giving birth without receiving prenatal care was compared over time at the state and regional level for various characteristics of the mothers and infants. These characteristics include the mother's age, race, marital status, and education, and the infant's birth weight. The results are interpreted along with the expectations that the percentages of women giving birth without prenatal care should be declining over time. Specific information can be identified about where, and for whom, these percentages are increasing. PMID- 10474983 TI - Can low volume liver transplant centers be successful? The Regional Transplant Center of Willis-Knighton & Louisiana State University Medical Center. The first 120 liver transplants. AB - The development of orthotopic liver transplantation represents this century's most significant advance in the management of liver disease. In the 1980s the vast majority of liver transplants were performed at several large centers; however, in this decade, improvements in techniques and success rates have allowed live transplantation to expand to regional centers across the country, particularly in the southeast. This proliferation of regional centers and the widening disparity between organ availability and numbers of recipients have created tremendous controversy at the national level regarding the allocation scheme used to distribute livers to recipients. The large programs today are advocating change to a national waiting list which would eliminate local priority and jeopardize the existence of smaller regional centers. Furthermore, the large programs favor establishing a limited number of megacenters where all liver transplants would take place, arguing that low volume centers cannot perform liver transplants with acceptable complication and survival rates. At the Regional Transplant Center of Willis-Knighton Hospital and Louisiana State University Medical Center in Shreveport (WK/LSUMC) we performed 122 liver transplants between July 1, 1991 and December 31, 1997. The purpose of this study was to examine our complication and survival rates and compare them to national averages. The actuarial graft survival at 1, 2, and 3 years in this series compared to the national average respectively was 76% and 70%, 66% and 66%, 62% and 62%. The actuarial patient survival (WK/LSUMC vs National) at 1, 2, and 3 years was 80% and 80%, 75% and 75%, 70% and 74%. The rate of retransplantation was 8% with a national average of 10% to 20%. Our rate of graft primary non function was 5% with the national average being 2% to 10%. The rate of vascular thrombosis of the graft in this series was 2% with a national rate of 5%. The differences in these results were not statistically significant (P < .05). Low volume transplant centers can perform liver transplant successfully, allowing the regionalization of the treatment of choice for end-stage liver disease. PMID- 10474984 TI - Radiotherapy of acral lentiginous melanoma of the foot. AB - Four previously unreported cases of acral lentiginous melanoma of the foot treated by radiation are reported. All patients, for a variety of reasons, were not suitable for complete surgical excision. Three large doses of radiation were employed using the 0, 7, 21 technique. Local response of the melanoma to radiotherapy was excellent and all patients were well palliated by the treatment. This treatment of acral lentiginous melanoma is recommended in patients with this diagnosis who, due to age, medical condition, or the presence of metastasis, are not suitable for standard treatment (surgery). PMID- 10474985 TI - Personality types in academic medicine. AB - Based on Swiss physician-scholar Carl G. Jung's theory of psychological types proposed in the 1920s, Kathleen Cook Briggs and her daughter Isabel Briggs Myers developed the Myers-Briggs Type Indicator (MBTI) three decades later. They applied Jung's dynamic theory to determine how persons take in information, make decisions, and communicate thoughts and feelings. Medical students were of special interest to their research and much has been written since then about the use of the MBTI in medicine. In this study, results of MBTIs administered to 1797 freshmen students at Louisiana State University School of Medicine--New Orleans from 1988 to 1998 are compared with those reported by the MBTI developers and others over the years and throughout the United States. Findings indicate some noteworthy shifts in the psychological profile of medical students over time and among schools that may be due to changes in the delivery of health care, the increase in technology in the practice of medicine, and the dramatic increase of women in medicine. PMID- 10474986 TI - The epidemiology of stroke in Mississippi and the United States. AB - Given current trends, Mississippi will not come close to meeting the National Health Promotion and Disease Prevention Objectives for the Year 2000 for reducing stroke mortality (Table). Without major population-based public health initiatives, stroke will remain a leading cause of morbidity, disability, and mortality and a major source of health care costs in the state for the foreseeable future. PMID- 10474987 TI - Endoscopic treatment of inverted papilloma. AB - Endoscopic resection of inverted papilloma involving the nasal septum and lateral wall of the nose is indicated in selected cases. Careful endoscopic monitoring of the patient every 2-3 months is mandatory. Traditional surgery is reserved for more extensive lesions, recurrent lesions, or patients who have developed squamous cell carcinoma arising from an inverted papilloma. An important part of the procedure is careful informed consent. The patient should be made aware of the possibility of recurrent lesion and the need for more extensive surgery in the future as the result of any recurrence. PMID- 10474988 TI - The year 2000 problem: guidelines for protecting your patients and practice. American Medical Association. PMID- 10474989 TI - Confidentiality and privilege of physician peer review. PMID- 10474990 TI - Endodontists are people, too. PMID- 10474991 TI - Court rules dentists to be judged by standards of honest marketplace. PMID- 10474992 TI - Natural remedies recommended for the management of oral health. AB - This paper discusses various practices recommended by homeopaths, herbalists, spiritual and natural healers for the management and maintenance of oral health. It is intended as a partial guide to educate the dental professional on self administered, over-the-counter remedies that are easily available to the general population. Since few if any clinical studies exist assessing the efficacy or side effects of these ingested products, little is known about the beneficial or potentially harmful course these remedies may take on the human body. PMID- 10474993 TI - Current periodontal plastic procedures around teeth and dental implants. AB - Periodontal plastic procedures are performed to prevent or correct anatomical, developmental, traumatic or plaque-induced defects of the gingiva, alveolar mucosa or bone. The majority of these procedures are performed in combination with restorative and/or orthodontic therapy, with the primary goal of enhancing esthetics. In this review, some of the more prominent techniques currently available to address mucogingival deficiencies, including pedicle grafts, free soft tissue grafts and combination grafts, are illustrated. In addition, potential complications associated with periodontal plastic procedures will be discussed. PMID- 10474994 TI - Caries and dentin. Myths & consequences. AB - The dentin-pulp complex is sensitive to infection. Restorative treatment should function as a Band-Aid protecting the dentin-pulp complex from bacteria and their byproducts. This article describes different ways of treating the exposed dentin pulp complex in such a way that bacterial infection can be avoided. PMID- 10474995 TI - [Progress in the study of tubulo-interstitial nephropathies]. PMID- 10474996 TI - [Structures and functions of the renal tubules and interstitium]. PMID- 10474997 TI - [Immunological mechanisms involved in renal tubular and interstitial lesions]. PMID- 10474998 TI - [Pathology of tubulo-interstitial nephropathies]. PMID- 10474999 TI - [Renal tubular dysfunctions and clinical symptoms of tubulo-interstitial nephropathies]. PMID- 10475000 TI - [Etiology and physiopathology of acute and chronic tubulo-interstitial nephritis]. PMID- 10475001 TI - [Tubulo-interstitial lesions in glomerular diseases]. PMID- 10475002 TI - [Idiopathic tubulo-interstitial nephritis]. PMID- 10475003 TI - [Infectious tubulo-interstitial nephritis]. PMID- 10475004 TI - [Drug-induced tubulo-interstitial nephropathies]. PMID- 10475005 TI - [Toxic tubulo-interstitial nephropathies]. PMID- 10475006 TI - [Tubulo-interstitial nephropathies caused by electrolyte disturbances]. PMID- 10475007 TI - [Metabolic tubulo-interstitial nephropathies]. PMID- 10475008 TI - [Radiation nephritis]. PMID- 10475009 TI - [Mechanism of the development of tubulo-interstitial nephropathies due to collagen diseases, angiitis, or granuloma and its significance]. PMID- 10475010 TI - [Tubulo-interstitial nephropathies caused by obstructive nephrosis]. PMID- 10475011 TI - [Mechanism of the development of tubulo-interstitial nephropathies caused by proteinuria]. PMID- 10475012 TI - [Clonality of infiltrative T cells in tubular dysfunction caused by Sjogren's syndrome]. PMID- 10475013 TI - [Clinical significance of tubulo-interstitial lesions and therapeutic strategies: discussion]. PMID- 10475014 TI - [Case of immunocytoma 4 years after and immunoblastic lymphoma 6 years after complete remission of Hodgkin's disease]. PMID- 10475015 TI - [Case of glomerular nephritis with C1q uniquely deposited in the mesangial region]. PMID- 10475016 TI - [Autopsy case of sudden death due to acute diffuse pulmonary thromboembolism that was caused by giant uterine myoma]. PMID- 10475017 TI - [Case of anaphylaxis caused by tiquizium bromide (Thiaton)]. PMID- 10475018 TI - [Case of immunotactoid glomerulopathy (ITG) associated with acute active glomerular lesion]. PMID- 10475019 TI - [Long-time survival of a patient with malignant pleural mesothelioma with infiltration of many plasmocytes as a unique feature]. PMID- 10475020 TI - [Clinical application of the new action of macrolide preparations to respiratory diseases]. PMID- 10475021 TI - [Current status and management of home dialysis programs]. PMID- 10475022 TI - [Prevention of hepatic carcinogenesis by retinoids]. PMID- 10475023 TI - [Role of the government for development of umbilical cord blood bank system in Japan]. AB - In Japan, development of umbilical cord blood transplantation is still on its preliminary stage. However, the nation-wide requests for dissemination of cord blood is growing. The Ministry of Health and Welfare established the "Committee on Umbilical Cord Blood Transplantation" last year and the committee has made the action plan to develop the nation-wide cord blood bank system. Our society is going to start the collaborative projects cooperated with medical professionals, politicians and community people. PMID- 10475024 TI - [Expectation of cell therapy in the 21st century]. AB - The era of stem cell transplantation is now moving from PBSCT to umbilical cord blood transplantation. In compensate demerits of those therapies, another new way would be found for the next generation. For instance, utilization of more immature stem cells with multi potential hematopoietic activity, combined transplantation of functional lymphocytes like natural killer cells for prevention of GVHD, effective introduction of HSV-tk gene, standardization for cell processing, and establishment of international cooperative center for cell processing. We will be needed to consider the genome therapy in the next century. PMID- 10475025 TI - [The current status and future aspects of umbilical cord blood banking system in Japan]. AB - Since 1995, when the first, non-governmental umbilical cord blood bank (CB bank) was created in Kanagawa prefecture, 9 local CB banks have been established in Hokkaido, Kanto, Tokai, Kinki and Kyusyu district. According to the experiences of organizing these local banks and of collecting the data of cord blood cell transplantation by 2 grant study groups, The Ministry of Health and Welfare is now creating national cord blood banking system, which will be opened in the coming April. PMID- 10475026 TI - [Collection of umbilical cord blood to separate the peripheral blood stem cells during the third stage of labor]. AB - In order to collect the umbilical cord blood for the separation of peripheral blood stem cells, obstetrician have to get the document of informed consent from the pregnant women for the umbilical cord blood donation during pregnancy. After the fetal delivery, umbilical cord blood is easier collected from the umbilical vein of the placenta in utero than that of the delivered placenta. After the sterilization of umbilical cord, the special bag attached with tube and needle is more useful to collect the blood disinfectively and abundantly than the syringe. Blood stem cells should be separated as soon as possible after collection, but umbilical blood may be stored in the refrigerator until 24 hours after the collection, if the immediate separation is impossible. There is the positive correlation between the collected blood volume or the preserved hours until the separating management and the separated stem cells. Hearing on the health of the child should be followed until 6 months after delivery. PMID- 10475027 TI - [Harvest techniques for cord blood after delivery of placenta]. AB - We describe harvest techniques for cord blood after delivery of the placenta. The umbilical vein is punctured or catheterized, and then placental/cord blood is aspirated with syringes contained anticoagulants. Other method of collection, which has been undertaken in the Cord Blood Bank of the New York Blood Center, is to drain blood into a collection-bag by gravity. The collection bags with anticoagulants are manufactured by TERUMO and BAXTER HEALTHCARE CO. PMID- 10475028 TI - [Quality control of cryopreserved cord blood cells for transplantation]. AB - Quality control (QC) of cryopreserved cord blood (CB) cells is very important for the safety of transplantation. In this article, the cell processing of CB cells, the laboratory tests for CB specimens, and the system of QC in CB bank is discussed. PMID- 10475029 TI - [Current status of cord blood banks in the world]. AB - Since the first successful transplantation of placental/umbilical cord blood (PCB) in 1988 at St. Louis Hospital in Paris, the effort has been made to establish large scale cord blood banks for allogeneic transplantation around the world. Recent reports from New York Blood Center which stocks 9000 units and shipped 700 units to US and other countries indicates that PCB transplantation is as effective as bone marrow transplantation. There are many cord blood banks in the world now and the number of PCB units stocked in the world are increasing rapidly. The methods for collection and processing of PCB, and test items for screening for viral infections, etc. differ greatly among banks. Therefore, the quality of PCB units stocked worldwide has become an important issue and some major banks are trying to introduce GMP or ISO system for the quality assurance of PCB. In this chapter, we introduce the activities of major cord blood banks in the world. Guidelines or regulation for hematopoietic stem cell processing including PCB has been enacted in some countries though the situation of support from their governments differ. International registry for PCB transplantation have been developed by EUROCORD. The international network of cord blood banks, NETCORD, has been established recently with major banks in Europe, United States and Japan to ensure the rapid supply of safe and efficient PCB units to the patients throughout the world. Obtaining information of international activities of cord blood banks in the world and cooperation with them should bring the National Cord Blood Bank in Japan into concordance with a developing world standards. PMID- 10475030 TI - [Clinical results of cord blood transplantation]. AB - Nearly 1000 cases have received cord blood transplantation in the world since the first successful transplantation by Gluckman. 79 cases have been reported to the Ministry of Health in Japan. Results of sibling transplantation have been good and the survival rates are as good as those in bone marrow transplantation. On the other hands, results of unrelated transplantation seem to be poor due to lower engraftment rate and higher transplant-related mortality. Number of attempts in adults is increasing, but the initial results do not look so good, probably because many transplants were done at the advanced stages, specially in adults. Further analysis and follow-up are needed for more accurate evaluation of this treatment modality. PMID- 10475031 TI - [Cord blood transplantation for adults]. AB - Current reports from Europe and USA show that unrelated cord blood transplantation (CBT) gives good results in treating with children with malignancies and other disorders. However, there are not enough patients in the adults to perform a comparison with other stem cell transplantations. In unrelated CBT, HLA disparity is not a limiting factor and the incidence and the severity of acute and chronic GVHD is lower, even in the state of HLA 2 or 3 antigen-mismatched transplant, than in other stem cell transplantation, but the number of cells infused is important. At this stage, unrelated CBT is an optional source of stem cells in adults patients with malignancies who lack HLA compatible donors in bone marrow donor registries. However, the choice of cord blood units containing large number of cells and good disease status at transplantation might improve results. PMID- 10475032 TI - [Cord blood transplantation and ex vivo expansion of hematopoietic stem cells]. AB - Umbilical cord blood has provides an alternative source to bone marrow for transplantation in children and some adults. However it has been thought to be necessary to expand hematopoietic stem cells in cord blood for adult-size transplantation. Flow-cytometric analysis revealed that most immature hematopoietic progenitors express gp130 but not interleukin-6 receptor (IL-6R). We established an ex vivo expansion system of human hematopoietic stem/progenitor cells using soluble IL-6R (sIL-6R). A combination of sIL-6R/IL-6 complex and SCF expanded CFU-Mix approximately 60-fold in both serum-containing and serum-free cultures by day 14. Addition of anti-gp130 mAbs and anti-IL-6R mAb to the above cultures dose-dependently inhibited the expansion of progenitors, suggesting that gp130 signalling initiated by the sIL-6R/IL-6 complex is important for significant expansion of human hematopoietic stem/progenitor cells. Addition of thrombopoetin and/or Flk2/Flt3 ligand (FL) to culture with sIL-6R/IL-6/SCF augmented the expansion of not only hematopoietic progenitors assayable in clonal culture but also hematopoietic stem cells estimated by NOD/SCID mice. Recent evidence shows that hematopoietic stem cells first occur and expand in aorta gonad-mesonephros(AGM) region at 10 to 11 days post coitum(dpc) in murine, which suggests that AGM region at this stage provides a microenvironment suitable for development for hematopoietic stem cells. We reported here on a novel stromal cell line derived from the AGM region at day 10.5 dpc, which supported for 6 weeks generation of human multipotential hematopoietic progenitors from cord blood CD34+38- primitive hematopoietic cells in a co-culture system. This cell line is expected to elucidate molecular mechanisms regulating early hematopoiesis, and pave the way for developing strategies for ex vivo expansion of human transplantable hematopoietic stem cells. PMID- 10475033 TI - [Peripheral blood stem cell transplantation--practices of mobilization and harvest]. AB - Recently, mobilized peripheral blood stem cells (PBSC) are increasingly used as an alternative to bone marrow for engrafting procedures. Chemotherapy followed by recombinant human granulocyte-colony stimulating factor (rhG-CSF) or rhG-CSF alone are the most commonly used PBSC mobilization schedules. Current timing of apheresis has now been yet decided on time by flowcytometric analysis of CD34 positivity of peripheral blood whole white cell count. Because apheresis procedure has been established quite well, PBSC harvest procedure becomes fast, safe and stable. PMID- 10475034 TI - [Preservation, thawing and infusion of hematopoietic stem cells]. AB - Peripheral blood stem cells can be stored by the following 3 methods: liquid storage, non-rate controlled freezing and rate controlled freezing. Methods of processing of these cells including thawing, ex vivo purging and infusion are described in detail. PMID- 10475035 TI - [Ex vivo and in vivo purging for PBSCT]. AB - Relapse due to either residual host disease or reinfused tumor cells remains the principal cause of treatment failure after autologous stem cell transplantation. It is very attractive to remove contaminated tumor cells from autologous grafts prior to transplant and many purging techniques including hyperthermia we developed, have been reported on this subject. However, there are only limited data suggesting that purging autografts have any favorable effect on relapses or disease-free survival. Drug-treated purging and CD34 positive cell selections that remove substantial numbers of T cells or destroy stem cells may have adverse effects such as delayed hematopoietic or T cell recovery. Therefore, there is a critical need for large, well-designed trials to re-evaluate ex vivo and in vivo purging effects on relapses and disease-free survival after autologous stem cell transplantation. PMID- 10475036 TI - [Allogeneic peripheral blood stem cell transplantation]. AB - Allogeneic peripheral blood stem cell transplantation (allo-PBSCT) has been increasingly used as an alternative to allogeneic bone marrow transplantation (allo-BMT). Medication of granulocyte colony stimulating factor (G-CSF) and apheresis are well tolerated by donors and supply adequate numbers of stem cells for the engraftment. Patients engraft sooner using PBSCT compared to allo-BMT. Allo-PBSCT is a safe alternative to allo-BMT and has distinct advantages for donors and patients. Faster engraftment results in fewer transfusion, shorter hospitalization, and decreased cost. However future research to determine if long term side effects from G-CSF will negatively affect donors is essential. Data regarding durability of hematopoiesis and incidence for graft versus host disease warrant further analysis. PMID- 10475037 TI - [PBSCT and GVHD]. AB - Allogeneic peripheral blood stem cell transplantation (PBSCT) has been increasingly used as an alternative to allogeneic bone marrow transplantation. Allo-PBSCT can provide rapid engraftment of neutrophils and platelets. Although the recipients of allogeneic PBSCT are infused 10-fold T cells compared with BMT, there is no evidence for a significant difference between PBSCT and BMT with regard to incidence and severity of acute graft-versus-host disease (GVHD). On the other hand, several reports have indicated a high risk for developing chronic GVHD after allogeneic PBSCT as opposed to BMT. PMID- 10475038 TI - [DNA typing methods of HLA]. AB - Human leukocyte antigens (HLA) are a group of cell membrane proteins encoded by the HLA gene complex on the short arm of the chromosome 6, with a high degrees of polymorphism, and a total of about 650 alleles have been identified at HLA-A, B, C and DR loci up to now, which regulate the immune response by self/not-self discrimination. A number of DNA-based typing methods using specific primers, specific probes or restriction enzymes have been developed. In HLA allele typing, a few methods which are simple in handling and able to determine the alleles automatically, have been available. PMID- 10475039 TI - [Serological typing of HLA]. AB - Micro-lymphocyte cytotoxicity test (LCT) have been developed by Dr. Terasaki at 1964 and usually used for serological typing up to date. HLA typing has been complicated by the extensive diversity of and the sequence homology among HLA alleles. However, it is need to match HLA types between donor and patient for hematopoietic stem/progenitor cell transplantation, therefore accurate HLA typing is required. Recently, it is able to perform accurate lymphocyte separation and HLA typing of much blood samples by using immunomagnetic beads and computer analysis, respectively. PMID- 10475040 TI - [Tests for donors of cord blood and bone marrow]. AB - To date blood transfusion is considered to be one of tissue or organ transplantation. In the hematopoietic stem cell transplantation, donors of bone marrow, peripheral blood stem cells (PBSC) or cord blood should be tested to keep safety of donors and patients. Donors of PBSC, cord blood and bone marrow should be selected on the results of HLA class I and class II. Furthermore, donors of bone marrow should be tested for general anesthesia in order to keep safety of donors. In the present paper, we described the lists of tests for patients and donors of blood, PBSC, cord blood and bone marrow before and after stem cell transplantation. PMID- 10475041 TI - [Detection of hematopoietic stem cells by flow cytometry, hematology analyzer or in vitro culture method]. AB - We describe 3 different ways to determine the number of hematopoietic stem or progenitor cells in peripheral blood and harvest juice for peripheral blood stem cell transplantation. Flow cytometric analysis of CD34 positive cells gives us relatively quick and reliable results, although it's quite expensive. Automated hematology analyzer, SE-9000, exhibits real time appearance of stem cell population in a least expensive way. Colony formation in vitro such as CFU-GM is a direct way to detect hematopoietic committed cells, however this is time consuming. We have to choose proper ways for each institute. PMID- 10475042 TI - [Screening test for hematopoietic stem cells in umbilical cord blood by automated hematology analyzer, SE-9000]. AB - An IMI channel on automated hematology analyzer SE-9000 was used for detection of stem cells in cord blood in comparison with other standard methods such as flow cytometry analysis and progenitor assays. IMI count in cord blood samples correlated with the numbers of CD34+ cells, CFU-GM and BFU-E. Also the cell count in HPC area showed a correlation with CD34+ cells number, but to lesser extent than IMI count, and there was no significant correlation with the numbers of progenitor cells. Cord blood contains higher frequency of immature subclasses of progenitor cells, and it might explain the difference in the result of HPC module between cord blood and peripheral blood stem cell sample. The results demonstrated a usefulness of the measurement of IMI-positive cells to estimate stem cell content in cord blood samples as a screening test for cell banking. PMID- 10475043 TI - [Future development of cord blood transplantation]. AB - In the present paper, we described disadvantages and advantages and future state of cord blood transplantation. One of most important disadvantages is a limited number of nucleated cells obtained from cord blood for transplantation. In order to overcome with the limited number of the cells, ex vivo expansion is considered to be essential for cord blood cell transplantation to adult patients. We also should develop the techniques of antigen presenting cells expanded ex vivo such as dendric cells pulsed with tumor specific antigen. Furthermore, we should create and supply unique cells from cord blood using genetic manipulation for the treatment of hematological disorders such as red cells without any red-cell antigen those can be transfused the patient with any blood type. PMID- 10475044 TI - [Thoughts about the domestic dog as the catalyst for relations between humans and a body contact object for humans]. AB - Based on an intensive analysis of literature, the study summarizes for the first time important but seldom realized reasons for dog keeping by humans. Firstly, the significance of the companion animal domesticated dog as catalyst of numerous different social contacts and communicative interactions of the dog keepers is emphasized. Secondly, the problem of total devotion of dog owners to the animal is discussed including its negative aspects, and the psychogram of a "typical dog owner" is described. Thirdly, the role of the domesticated dog as object of body contact (petting object) for humans is characterized, i.e., the positive effects of tactile stimuli for the emotional and physical well-being of dog owners as well as their animals are presented. All themes are finally discussed in the light of the historical dimension of dog domestication, whereby it is obvious that one of the first uses of this mammalian species was that as an object of affectional touch and hugging to serve basic human emotional needs. In this way it becomes also evident how deeply the dog depends on its owner. PMID- 10475045 TI - [Chlamydia abortion in sheep: possibilities for serological diagnosis using a competitive ELISA and insight into the epidemiologic situation in Switzerland]. AB - 466 sheep sera out of 19 flocks in Switzerland were examined by a competitive enzyme linked immunosorbent assay (cELISA) for antibodies against Chlamydia psittaci "serotype 1" ("ovine enzootic abortion"). Since numerous positive reactors were found in flocks without abortion history, 30 fecal samples out of two of these flocks were examined by PCR for evidence of chlamydial DNA. One of these samples turned out to contain DNA of Chlamydia psittaci "serotype 1". These results suggest, that in Switzerland "serotype 1" of Chlamydia psittaci is widespread not only as cause of chlamydial abortion but also as latent intestinal infection in sheep. The resulting difficulties for serological diagnosis of chlamydial abortion and possible solutions based on the cELISA are discussed. The complement fixation test (CFT), still considered as standard method for serological examination for Chlamydiae, has additionally been applied. PMID- 10475046 TI - [Case presentation from small animal medicine. Diabetes mellitus in a cat]. AB - This case report is about a 9 year old female spayed domestic shorthair cat with a previously diagnosed diabetes mellitus. After an insulin treatment for a short period of time, the cat was given an oral hypoglycemic agent. Four years after initiating therapy the cat was presented with diabetic ketoacidosis. Various aspects of the etiology, pathophysiology and treatment of diabetic ketoacidosis are discussed. PMID- 10475047 TI - [Mode of onset and two years outcome of a broad spectrum of affective disorders]. AB - In recent years emphasis has tended to be placed on assessing and diagnosing low grade affective disorders. If mild affective symptoms are also considered a broad spectrum of affective disorders, this spectrum can be seen, not only in the manifestation of major symptoms of such illnesses as manic-depressive illness, but also in symptoms of various mental disorders. The present study was undertaken to analyze the clinical features of a broad spectrum of affective disorders in a prospective follow-up study. Ninety mentally ill patients who visited our outpatient clinic were followed for 2 years, to investigate the mode of onset, the course and outcome of their disorders. Using a semistructured interview method specially developed for this survey, these patients were divided into a broad spectrum of affective disorder group and the other groups. Forty nine patients were allocated to a spectrum of affective disorder group characterized by four-day (or longer) persistence of at least one of the following symptoms, depressed mood, loss of interest or elevated, expansive, or irritable mood. Of the forty-nine patients there were only two patients with manic symptoms. And the forty-seven patients with depressive symptoms were compared with the other groups as a main subject. The other groups were composed of eighteen patients with psychotic symptoms (the psychotic group) and 23 patients with neurotic symptoms (the neurotic group). The psychotic and neurotic groups did not satisfy the criteria shown above. There was no significant differences in male-to-female ratio or age between these three groups. When the mode of onset of symptoms were compared, the percentage of cases in whom symptoms could be identified early (within 10 days after onset) was 64.4% in the depressive group, 41.2% in the psychotic group and 30.4% in the neurotic group. The percentage of cases who remitted 2 years later was 70.2%, 38.9%, 45.5% in the depressive group, psychotic and neurotic groups, respectively. The GAS score (mean +/- SD), assessed 2 years later, was 76.2 +/- 12.5, 62.8 +/- 11.7, 78.2 +/- 9.9 points for these three groups, respectively. These results suggest that a broad spectrum of depressive disorders develops more acutely and patients with this spectrum are more likely to recover from symptoms and in function. Thus, the course of disorders was more favorable in the depressive group than in the psychotic group or the neurotic group. PMID- 10475048 TI - [In Process Citation] PMID- 10475049 TI - [Chaucer and the language of science]. AB - The study of the medieval sciences and their technical vocabulary has been considered essential to an adequate understanding of Chaucer's works ever since the nineteenth century. But any attempt of evaluating Chaucer's scientific vocabulary is confronted with several major difficulties. The well-known fact that the Norman Conquest caused the breakdown of an established literary standard and eventually led to the dominance of French and the suppression of the native vernacular as a means of instruction and formal education up to the middle of the fourteenth century is equally responsible for the comparative lateness of Middle English literature on the artes. This limitedness of comparative material also contributes to the difficulty in evaluating Chaucer's scientific lexicon and its use in fictional contexts. The still unsatisfactory state of scholarship in the field of Middle English prose enhances these problems. Concerning Chaucer, three separate possibilities are discussed: the poet follows an already established native tradition, he makes use of existing French or Latin terminology, or he coins a new word. Especially the third case poses intricate problems which are connected with the question of the status of words, since it is often impossible to draw precise limits between different categories.--In addition to that, Chaucer's use of the hermeneutic technique of etymology and the linguistic importance of variant readings are dealt with. PMID- 10475050 TI - ["Medicinarn aspernari impietas est."--On the relationship of the Reformation and academic medicine in Wittenberg]. AB - Philipp Melanchthon's university reform at Wittenberg which was due to Reformation mainly concerned the Arts faculty, but also affected the medical education. Influenced by Melanchthon's orations about medical subjects and by his textbooks on natural philosophy and anthropology, Wittenberg medicine was taught within the theological framework of evolving Protestantism. This was particularly true for the anatomical education: In the context of the protestant distinction between gospel and law anatomy was considered as part of the law. Anatomical knowledge was therefore regarded necessary for all students, not only for those becoming physicians. Furthermore it seems that the adoption of new anatomical findings (e.g. Vesal's corrections of Galen) was more promoted than hindered by the theological context of Wittenberg anatomy. PMID- 10475051 TI - [The physicians of Friedrich von Hardenberg and Sophie von Kuhn]. AB - Relatively little is known about the physicians who attended the German writer Friedrich von Hardenberg (1772-1801) and his fiancee Sophie von Kuhn (1782-1797) during the years immediately preceding their deaths. An attempt is made here to gather more information about these doctors on the basis of archival and other neglected sources. However, due to the fragmentary nature of the evidence it cannot be determined what, if any, connections exist between the treatment methods of these physicians and Hardenberg's thinking on medicine. PMID- 10475052 TI - [Friedrich Wilhelm I and porphyria]. AB - Historical evidence has been collected attempting to diagnose members of royal houses, perhaps most publicized by Macalpine and Hunter (1969) for George III and his assumed porphyria, claiming that his insanity was a classic case of thereof. This rare metabolic disease presents with a variety of signs and symptoms: skin disease, abdominal pain, tachycardia, and neuro-psychiatric findings. The porphyrias are hereditary and since George III and Frederick William I share ancestors it seemed reasonable to investigate if the latter may also have suffered from porphyria. The pathography of both kings is meticulous, showing for both that abdominal pain, erratic behavior, restlessness, and discolored urine were frequently observed and complete recovery interictally was common. Intercurrent illnesses, fasting, alcohol and even tobacco smoking have been shown to be inducers of attacks and these risk factors are well documented in royal history.--The diagnosis of porphyria was not recognized then and other names were used, such as Cachexia hypochondriaca, Asthma spasmodico flatulentum, dolores arthritici.--We propose that Frederick William I suffered from an inducible porphyria. PMID- 10475053 TI - [Galen on the topic of abnormalities]. AB - Galen on deformities--Galen's treatment of problems of deformities has its primary source in his twofold teleological conviction that nature acts purposefully and that the human organism and his parts serve their purposes in an optimal way. Thus he defines deformities as functionally deviant body parts. Operative therapy therefore is indicated only if an abnormal part of the body hinders its normal function. On the basis of scientific physiological concepts of his time Galen blames avoidable diatetic failures of the parents as causes of deformities. Galen's engagement on this point reveals the influence of his own experiences as a physician. Galen's statements on deformities show that his teleological concept of nature, his practical medical experience and scientific theories of his time enable him to draw fruitful theoretical and practical conclusions for matters of definition, indication for treatment and prophylaxis of deformities. PMID- 10475054 TI - ["This helps; this is good". Horse books of Tibetan Himalayas]. AB - The present paper summarizes our recent investigations of the so-called horse books from the High Himalayas in Nepal. These books are written in tibetan language and are essentially dealing with hippology and hippiatry and to a lesser extent with topics such as pharmacology, anatomy, methods of diagnosis, divination and magical practices for horse races. The therapeutic methods of treatment in tibetan veterinary medicine are guided by the concepts of human medicine which, on the one hand, are related to the Ayurvedic System, on other hand, to the Traditional Chinese Medicine. Concerning the structure of these manuscripts a lot of similarities could be observed with corresponding treatises on horse management and medicine from Europe. PMID- 10475055 TI - [Handwritten documents of 'Antidotarius magnus']. AB - The 'Antidotarius magnus'--compiled about 1080 by the archbishop of Salerno, Alphanus--deals with the pharmacological methods of healing and contains nearly 1073 antidots. We have discovered 13 Latin manuscripts of the 'Antidotarius magnus' in the libraries of Basel, Bern, Cambridge, Erfurt, Florence, London, Oxford, Paris and Parma. One should remember that the MS Taurin.I.VI.24 had been totally destroyed in 1904, but this manuscript is still noted in the catalogues of Pasin, Giacosa and Thorndike/Kibre. The most remarkable manuscript--the MS Palat.lat.747--is preserved in the National Library at Florence. The MS Palat.lat.747 is dated to 1153 and seems to be similar to the archetype. We are currently preparing a critical edition based on the oldest manuscripts. PMID- 10475056 TI - [Antitumor compounds isolated from higher plants]. AB - A lot of anticancer agents have been isolated from natural sources, especially from microorganisms and plants. However, there is no special type of compounds for cancer therapy. Various types of substances are effective for various types of cancers and tumors: for instance, alkaloids, lignans, terpenes and steroids, etc. In this report, the authors will describe especially about higher plants. PMID- 10475057 TI - [A cytoprotective chloride channel in gastric parietal cells]. AB - This review summarizes the regulatory mechanisms and physiological functions of the novel sub-pS Cl- channel (0.3 pS) that is present abundantly in the basolateral membrane of rabbit gastric parietal cells. The sub-pS Cl- channel is voltage-independent and inhibited by NPPB, a Cl- channel blocker. We found that this gastric Cl- channel is linked to three important physiological roles. First, the sub-pS Cl- channel has a housekeeping role through dominating the cell membrane potential. Although several types of cation channels are present, they do not significantly contribute to the membrane potential in the parietal cells. Second, the Cl- channel is activated by prostaglandin E2 via the EP3 receptor/Ca2 /nitric oxide (NO)/cGMP pathway. A vasodilator ecabapide also activates the channel by increasing the intracellular cGMP content. The NO/cGMP pathway mediated opening of the sub-pS Cl- channel is essential for cytoprotection against ethanol-induced damage in the gastric parietal cells. The NO/cGMP elicited cytoprotection is abolished by NPPB. To our knowledge, this Cl- channel is the first identified target for the cytoprotective NO/cGMP pathway. Third, the sub-pS Cl- channel is inhibited by the GTP-binding protein-mediated intracellular production of superoxide anion. Hydrogen peroxide and hydroxyl radicals have no effect on the channel activity. The intracellular superoxide anion acts as a messenger in the negative regulatory mechanism of the sub-pS Cl- channel. The similar sub-pS Cl- channel is also found in rat gastric parietal cells. PMID- 10475058 TI - [Applications of analytical ultracentrifuge to molecular biology and pharmaceutical science]. AB - Analytical ultracentrifuges, XL-A and XL-I, developed by Beckman Company now find a broad application not only in universities but also in industries. Especially they are utilized conveniently in industries aiming at the development of proteins as a therapeutic drug or in those targeting drugs composed of small molecules developed on the basis of the structures of proteins. Sedimentation techniques can be used 1) to determine the molecular weight of proteins in solution, 2) to examine protein aggregation, 3) to evaluate the molecular shape of proteins, 4) to study the interaction of proteins, e.g. between ligands and receptors, and 5) to obtain insight into biological functions of homologous proteins. Application of this technique to molecular biology and pharmaceutical science will be reviewed with ample examples. PMID- 10475060 TI - The Association for Patient-Oriented Research (APOR): ameliorating the crisis in clinical investigation. PMID- 10475059 TI - [Molecular identification of cytokinin-specific binding protein]. AB - Synthetic phenylurea derivatives such as N-phenyl-N'-(4-pyridyl)urea (4PU) and N (2-chloro-4-pyridyl)-N'-phenylurea (4PU30) have strong cytokinin activities. Using tritiated 4PU30 as a probe, we found the presence of a cytokinin-specific binding protein (CSBP) with high affinity for 4PU30 (Ka for 4PU30 = 4 x 10(10) M 1) in the soluble fraction of etiolated mung bean seedlings. We purified CSBP by the use of 4PU-Sepharose 4B, an affinity gel ligated with 4PU. Analysis of its cDNA revealed that CSBP was a novel member of a major pollen allergen/pathogenesis-related protein family with a calculated molecular weight of 17 kDa. Recombinant CSBP was expressed in Escherichia coli was confirmed to bind specifically to cytokinins. PMID- 10475061 TI - The patched signaling pathway in tumorigenesis and development: lessons from animal models. AB - The identification of mutations in the human homolog of the Drosophila segment polarity gene Patched in basal cell carcinoma has sparked intense interest in the role of this gene in human disorders. The transmembrane protein Patched is a receptor for the morphogene Sonic Hedgehog. Sonic Hedgehog/Patched signaling involves another transmembrane protein, Smoothened, and its intracellular effectors, including the proto-oncogene GLI1. During the past 2 years it has become evident that mutations in Patched or in one of the components of its signaling pathway contribute to the formation of several common human tumors. It is now well established that Patched is a tumor suppressor gene. The Sonic Hedgehog/Patched/Smoothened signaling pathway is thus rapidly emerging as one of the most important regulators of oncogenic transformation. This pathway also plays an important role during mammalian embryonic development. This dual role is especially visible in humans with inherited Patched mutations. Such patients suffer from Gorlin, or nevoid basal cell carcinoma, syndrome and exhibit a variety of developmental defects accompanied by a predisposition to tumor formation. Activating mutations in Sonic Hedgehog and Smoothened lead to similar phenotypes as do loss-of function mutations in Patched. By means of transgenic and gene targeting technologies the respective mutations have been expressed in the mouse. Such mutant mouse strains exhibit many symptoms observed in humans. These strains are useful models to study the pathogenesis of several common human tumors and developmental defects. Furthermore they provide important tools to study the Sonic Hedgehog/Patched/Smoothened signaling at the molecular and biochemical level. PMID- 10475062 TI - Gene expression profiling, genetic networks, and cellular states: an integrating concept for tumorigenesis and drug discovery. AB - Genome-wide expression monitoring, a novel tool of functional genomics, is currently used mainly to identify groups of coregulated genes and to discover genes expressed differentially in distinct situations that could serve as drug targets. This descriptive approach. however, fails to extract "distributed" information embedded in the genomic regulatory network and manifested in distinct gene activation profiles. A model based on the formalism of boolean genetic networks in which cellular states are represented by attractors in a discrete dynamic system can serve as a conceptual framework for an integrative interpretation of gene expression profiles. Such a global (genome-wide) view of "gene function" in the regulation of the dynamic relationship between proliferation, differentiation, and apoptosis can provide new insights into cellular homeostasis and the origins of neoplasia. Implications for a rational approach to the identification of new drug targets for cancer treatment are discussed. PMID- 10475063 TI - The coactivators p300 and CBP have different functions during the differentiation of F9 cells. AB - We have characterized an element (differentiation response element, DRE) in the promoter region of the c-jun gene that is both necessary and sufficient for retinoic acid (RA) and adenovirus early region (E1A) mediated up-regulation of c jun gene expression during the differentiation of F9 cells. The DRF complex, which binds specifically to DRE, is composed of the E1A-associated protein p300 and the activation transcription factor-2 (ATF-2) as a DNA-binding subunit of the DRF. The molecular association of p300 and ATF-2 enhances the transcription of the c-jun gene, which requires protein kinase C alpha mediated phosphorylation of Ser-121 of ATF-2 within its p300 interaction domain. We used antisense oligodeoxynucleotides (AS-ODNs) capable of binding specifically to the mRNA for either p300 or CBP to examine the individual roles of p300 and CBP during the RA induced differentiation, exit from the cell cycle, and apoptosis of F9 cells. F9 cells treated with AS-ODNs specific for p300 mRNA became resistant to RA-induced differentiation, while cells incubated with AS-ODNs specific for CBP mRNA were still able to differentiate. Despite their similarities p300 and CBP appear to have distinct functions during the differentiation of F9 cells. These results suggest that ATF-2 and p300 cooperate in the control of transcription by forming a protein complex in response to RA or E1A, and that the phosphorylation of ATF-2 and p300 is probably a signaling event in the pathway that leads to the transactivation of the c-jun gene in F9 cell differentiation. PMID- 10475064 TI - The quest for novel bioactive peptides utilizing orphan seven-transmembrane domain receptors. AB - Various sorts of bioactive molecules including hormones, neurotransmitters, and chemokines transmit signals into cells by binding to so-called seven transmembrane-domain receptors (7TMRs). The recent progress in cDNA and genome DNA analyses has brought the discovery of numerous genes encoding ligand-unknown "orphan" 7TMRs. We have developed a strategy to identify the ligands of orphan 7TMRs by monitoring specific signal transductions induced in cells expressing orphan 7TMRs. Employing this method, we succeeded in identifying the natural ligands of the orphan 7TMRs, hGR3, and APJ. The ligand peptide identified for hGR3 was found to show a specific prolactin release promoting activity in rat anterior pituitary cells in in vitro culture and was therefore named "prolactin releasing peptide." We named another novel bioactive peptide "apelin," for "APJ endogenous ligand." Although the biological functions of apelin are still under investigation, APJ reportedly acts as a coreceptor in the process of human immunodeficiency virus infection. We believe that the identification of orphan 7TMR ligands will provide clues to reveal the unknown regulatory mechanisms of various physiological phenomena and opportunities for novel drug discovery in the future. PMID- 10475065 TI - Inverse regulation of vascular endothelial growth factor and VHL tumor suppressor gene in sporadic renal cell carcinomas is correlated with vascular growth: an in vivo study on 29 tumors. AB - Tumors associated with the VHL (von Hippel-Lindau) disease, such as hemangioblastomas and renal carcinomas and their sporadic counterparts, are cystic and well vascularized. Mutations of the VHL tumor-suppressor gene and elevated levels of vascular endothelial growth factor (VEGF) have been described in these tumors. The upregulation of VEGF has been shown in vitro as a consequence of alteration of the VHL gene. No comprehensive in vivo analysis has yet been carried out of the factors affecting tumor growth, vascularization, VEGF, and VHL expression. We performed immunohistochemistry and mRNA studies on primary sporadic renal carcinomas and matching normal renal tissue. We semiquantitatively analyzed 29 renal carcinomas (22 clear cell, 5 chromophilic, 2 chromophobic tumors) for VHL mRNA, and VEGF expression for morphology and tumor size. Immunohistochemistry was carried out for VEGF protein expression, vascularization, and macrophage infiltration. Vascularization of the chromophilic renal carcinomas was lower than that of the clear cell type of renal carcinoma. Low VEGF protein expression was seen in four of the five chromophilic renal carcinomas. We found two groups of clear cell renal cell carcinoma: one with reduced VHL mRNA and increased VEGF mRNA, and the other without significantly altered VHL or VEGF mRNAs. Tumor vascularization was correlated with VEGF protein and seemed to be independent of macrophage infiltration. Our in vivo findings support the inverse relationship between the regulation of VHL and that of VEGF. Our data also indicate that there may be an VHL-independent pathway for the induction of tumor vascularization. PMID- 10475066 TI - The Glu-298-->Asp (894G-->T) mutation at exon 7 of the endothelial nitric oxide synthase gene and coronary artery disease. AB - We examined associations between the endothelial nitric oxide synthase (eNOS) gene Glu-298-->Asp (894G-->T) mutation and the occurrence and severity of angiographically defined coronary artery disease (CAD). eNOS mediates basal vascular wall nitric oxide production, and altered nitric oxide production has been implicated in atherosclerosis. The newly identified eNOS Glu-298-->Asp mutation in exon 7 is common and likely to be functional. It was found to be associated with myocardial infarction (MI) in Japanese but not in whites. We genotyped 763 white Australians undergoing coronary angiography for the eNOS Glu 298-->Asp mutation. The frequencies of the eNOS GG, TG and TT genotypes were 47.8%, 41.2% and 11.0% in men and 45.2%, 41.1% and 13.7% in women with CAD, and were not significantly different from those without CAD (43.2%, 40.7% and 16.0%, P=0.423 in men; 40.2%, 48.1% and 11.7%, P=0.582 in women). The mutation was also not associated with MI (P=0.469 in males; P=0.389 in females) or with the number of significantly stenosed vessels (P=0.954; P=0.734). The "T" allele frequency (32.5%) was much greater than that reported for the Japanese population (7.8% in controls and 10.0% in MI patients). In conclusion, the eNOS Glu-298-->Asp mutation is common, occurring with an allele frequency of 32.5%, but is not associated with either the occurrence or severity of CAD in the Australian population or with other established coronary risk factors assessed in our study. The mutation is significantly more frequent in the Australian than in the Japanese. PMID- 10475067 TI - Are cardiac troponins the myocardial markers for the new millennium? PMID- 10475068 TI - Leucocyte activation markers in clinical practice. AB - The response against tissue injury and infection begins with the early activation of molecular and cellular elements of the inflammatory and immune response. Severe tissue injury, necrosis, and infection induce imbalanced inflammation associated with leukocyte over-stimulation and excessive or dysregulated release of cellular mediators. Clinical and experimental studies have shown that these mediators are directly related to progressive post-injury complications. Persistent increased levels of pro-inflammatory mediators produce tissue injury. Excessive production and activity of anti-inflammatory mediators cause anergy and/or immune dysfunction with increased susceptibility to infection. Leukocyte activation is assessed by cell surface phenotype expression, cellular mediators determination, or by measuring functional responses using isolated cells. Potential routine clinical uses are: evaluation of severity and prognosis in critically ill patients, immunomonitoring of sepsis, and detection of tissue injury, necrosis, and infection. In practice, the determination of cellular activation markers is restricted by a limited number of automated methods and by the cost of reagents. The availability of flow cytometry and immunoassay automated systems can contribute to a wider use in practice. Here we review the immunopathophysiology of polymorphonuclear neutrophil, monocyte, macrophage, and lymphocyte activation in response to tissue injury and infection. In addition, laboratory methods for their determination, and clinical applications in practice, are discussed. PMID- 10475069 TI - Hepatitis C virus-specific DNA sequences in human DNA: differentiation by means of restriction enzyme analysis at the DNA level in healthy, anti-HCV-negative individuals. AB - This study aimed to look for further HCV-specific sequences at the DNA level of healthy, HCV-negative individuals. Here, the sequence section from nt 57 to nt 328 within the 5'-NCR was assayed. Different combinations of primers were used in the nested PCR without a preceding reverse transcriptase step, resulting in fragments of the expected molecular weight size and also shorter and longer ones. It shows that the major part of the 5'-non-coding region (5'-NCR) of the hepatitis C virus genome is present in the DNA fraction from peripheral blood mononuclear cells (PBMCs) of healthy, anti-HCV-negative individuals tested. Furthermore we designed experiments to prove the specificity of these findings, by using restriction enzymes for digest assays of the target DNA before PCR (pre PCR digest) and of the products after PCR (post-PCR digest). In conclusion, our study indicates that the main part of the internal ribosome entry site (IRES) structure of HCV at least is contained in the DNA of the individuals tested. PMID- 10475070 TI - Characterization of monoclonal antibodies to human protein 1/Clara cell 10 kilodalton protein. AB - Human protein 1/Clara cell Mr 10,000 protein consists of two identical subunits of seventy amino acid residues each. In the present study, eight clones of monoclonal antibodies against native protein 1 were prepared and their respective epitopes were immunochemically and immunohistochemically characterized using native protein 1, truncated recombinant protein 1 and synthesized peptides. Among the clones, three designated as TY-5, TY-7 and TY-8 recognized amino acid residues 7-16, residues 19-28, and residues 39-46, respectively, all of which comprise the hydrophobic cavity of protein 1, possibly associated with chemical binding function. With the exception of TY-4, the remaining clones recognized residues 61-68 which are exposed to solvent. The epitope of TY-4 remains undetermined. Proper selection and combination of clones and recombinant protein 1 may be useful for fundamental and clinical studies of protein 1. PMID- 10475071 TI - Rapid analysis of parathion in biological samples using headspace solid-phase micro-extraction (HS-SPME) and gas chromatography/mass spectrometry (GC/MS). AB - A simple and rapid method for the analysis of parathion in biological samples is presented. The method consists of the extraction of parathion from blood samples by headspace solid-phase micro-extraction (SPME), followed by capillary gas chromatography and mass spectrometry detection. The recoveries in the blood samples after addition of ammonium sulphate and sulphuric acid were between 85% and 89% compared to samples prepared in water. Linearity was established over a concentration range of 0.1-5 microg/g blood with acceptable coefficients of correlation and limits of detection reached 0.02-0.05 microg/g. The time for an analysis is 57 minutes for one sample, including the extraction step. In conclusion, HS-SPME in combination with GC/MS is an effective method for the determination and quantification of parathion-ethyl and parathion-methyl in biological material. PMID- 10475072 TI - Semi-automated rapid isoelectric focusing of apolipoproteins C from human plasma using Phastsystem and immunofixation. AB - Apolipoproteins (apo) C-I, C-II, and C-III play crucial roles in intravascular lipid metabolism. Whereas apo C-II is an obligate cofactor for lipoprotein lipase, apo C-III was shown to inhibit its action. Apo C-I can be a potent cofactor of human lecithin:cholesterol acyltransferase. Structural mutants and deficiencies of apo C-II lead to hypertriglyceridemia. A similar phenotype is associated with apo C-III mutants and is inducible by overexpression of human apo C-III in transgenic animals. No structural variant has so far been reported for apo C-I. The present paper describes a rapid semi-automated procedure for isoelectric focusing analysis of these C-apolipoproteins from whole plasma or serum and their visualization by immunofixation and silver staining. The procedure allows detection of charged variants of C-apolipoproteins. As applied to 295 patients with coronary heart disease and 85 controls, it also serves to detect deficiency syndromes of these apolipoproteins. The procedure provides reliable, easy and quick analysis of C-apolipoproteins applicable as a routine or screening procedure not restricted to specialized laboratories. PMID- 10475073 TI - The c-erbB-2 oncogene amplification by competitive PCR in aneuploid cell clones flow sorted from breast cancer samples. AB - The amplification of c-erbB-2 oncogene has been reported to have clinical relevance as a prognostic index in breast cancer. However, controversies still remain about its interpretation, mainly due to the inaccuracy of methods used for this purpose and to the unpredictable variability of the ratio between cancer and normal cells. Accurate quantitative assay, combined with strategies for selection or enrichment of tumor cell populations, could shed a new light on the relationships between molecular alterations and their clinical relevance. In this study, amplification of c-erbB-2 was measured by competitive PCR in 21 aneuploid breast cancers using a multiple DNA competitor both in whole homogenized cancer cells and in aneuploid enriched clones obtained after flow cytometry cell sorting. Most breast cancers (10/12) carrying c-erbB-2 oncogene amplification showed a significant increase in copy number in sorted aneuploid clones, and 2/9 apparently not amplified in basal samples were found to be amplified after being sorted for the aneuploid population. A general concordance between amplification and c-erbB-2 overexpression was found. The mean degree of amplification in sorted aneuploid clones is increased in breast cancers with the highest levels of c-erbB 2 protein overexpression. These data indicate that in breast cancers the amplification of c-erbB-2 oncogene is mainly associated with aneuploid cells. PMID- 10475074 TI - The index of individuality is often a misinterpreted quantity characteristic. AB - The concept of the "index of individuality" was introduced by Eugene Harris in 1974. The index of individuality, calculated as (CV(A)2 + CV(I)2)(1/2)/CV(G), where CV(A), CV(I), and CV(G) are analytical, within-subject, and between-subject coefficients of variation respectively, has been used by many to investigate the utility of conventional population-based reference values. For a high index of individuality, > 1.4, it has been said that reference intervals will be more useful than for a low index, < 0.6. The validity of these concepts is investigated here and a number of our findings are at odds with the generally held opinion. The index of individuality has no impact on the fraction of individuals classified using population-based reference values, as long as the change in concentration from the usual state is of the same absolute magnitude and one sample is assayed to detect disease. However, when a measurement falling outside a reference limit is repeated in order to verify the finding, the index of individuality has considerable influence. For quantities with very low indices, the repeat test result, will be close to the first and give no new information, whereas for quantities with high indices, a repeat test will decrease the number of true positives and false positives. PMID- 10475075 TI - Assessment of package inserts for diagnostic kits. AB - To assure the quality of service in laboratory medicine, it is necessary to implement a quality system which comprises the entire testing process. The use of quality reagents is an important aspect of the process. Despite the fact that it is the responsibility of the laboratory to ensure the quality of the analytical system (including reagents) and since it is impossible to evaluate all commercial diagnostic kits, the laboratory often depends on statements issued by the manufacturer to select the most appropriate diagnostics for a particular laboratory. In this study we report the results of the analysis of information provided in 887 package inserts enclosed in the more widely used commercial diagnostic kits, following the Standard for the labelling of clinical laboratory materials of the European Committee for Clinical Laboratory Standards (ECCLS). Only a third of these were in agreement with the guidelines of ECCLS Standard, reporting complete and correct information. We believe that it is necessary to implement a constructive cooperation between manufacturers of diagnostic materials and clinical laboratories to produce a more uniform approach to improvements in laboratory quality assurance. PMID- 10475076 TI - External quality assessment in clinical chemistry: review of the situation in Croatia with particular reference to equipment. AB - The effect of the use of new flame photometers (Ciba Corning 480, Chiron, Salzburg, Austria) and multichannel analyzers (BM Hitachi 904 or BM Hitachi 911, both Boehringer GmbH, Mannheim, Germany) on analytical quality was assessed in a number of medical biochemical laboratories (n=58) at health centers and general hospitals that provide primary health care services for the respective catchment areas. The laboratories were supplied during 1996 and 1997 with new equipment, as part of the First Croatian Health Project, Primary Health Care Subproject, carried out by the Croatian Ministry of Health and Croatian Institute of Health Insurance. We evaluated analytical performance of these laboratories for 19 analytes, according to the results reported in the Croatian External Quality Assessment Program for medical biochemical laboratories, performed four times per year. Changes of percentages of particular methods used by the participating laboratories were observed for inorganic phosphate, total bilirubin, urea and creatinine. Such changes were even more pronounced for enzymes, where changes in the measurement procedure such as incubation temperature and buffers were observed in the methods used. Evaluation of method performance revealed that after the introduction of new equipment interlaboratory variation decreased, so that an increasing proportion of laboratories included in the Project produced results within the target limits. PMID- 10475077 TI - Point-of-care testing: the views of the working group of the Dutch Association of Clinical Chemistry. AB - There is clearly a demand for near-patient laboratory testing. Modern technological developments support its implementation, but these are not the only preconditions that need to be satisfied in order to successfully introduce decentralized testing. In part 2 of this paper we give a definition of decentralized testing. Part 3 presents the advantages and disadvantages. This list is a summary taken from the suggested literature. Part 4 examines the formal position of the clinical chemist. Of major importance to the situation in the Netherlands are the Care Institutions Quality Act and the Individual Health Care Professions Act. Part 5 discusses professional standards. Part 6 outlines practical considerations which might support the implementation of decentralized testing. The clinical chemist should play an active role in drawing up a discussion paper for his own hospital. PMID- 10475078 TI - Preliminary evaluation of the new hematology analyzer COULTER GEN-S in a university hospital. AB - The COULTER GEN-S system (COULTER Corp, Miami, USA) is an automated hematology instrument that is designed to provide a complete hematological profile including white blood cells (WBC), complete blood count (CBC) differential count (diff) and the reticulocyte parameters. It was evaluated in our laboratory over a one month period. A preliminary study was performed using the GEN-S software revision 1D. The evaluation had two purposes: 1) evaluation of the GEN-S specifications; 2) comparison of its analytical performance with the hematology analyzer currently used in our laboratory. The first part of the evaluation showed that the COULTER GEN-S is reproducible, has linearity beyond the specifications given by the manufacturer and produces stable results up to 48 hours after blood collection. The evaluation of analytical performance included: 1) a comparison between the GEN-S CBC and diff numerical results and the COULTER STKS (COULTER Corp, Miami, USA). These comparisons showed that the results given by the two methods are similar and suggested that the COULTER GEN-S could replace the current hematology instrument in use in our laboratory; 2) a performance analysis, to measure the system's ability to detect morphologic abnormalities, when compared to the reference blood smear examination. This part of the evaluation was performed on both normal and abnormal samples. The results of this analysis showed that the sensitivity of the GEN-S was excellent, especially regarding blast cells, immature granulocytes, nucleated red blood cells (NRBCs) and platelet clumps when using the complete suspect flagging system of the instrument. In addition, the use of the review criteria in our laboratory allowed to detect all hematological diseases. The overall false negative rate was 0.9%. Thus we consider that the COULTER GEN-S system is suited for use in medium-to large hospital laboratories which perform more than 100 CBC/day. Overall, the instrument had excellent performance, with a throughput of more than 100 samples/h. Its user friendly workstation has complete patient data management, which is in compliance with good laboratory practices in France. PMID- 10475079 TI - Use of biochemical markers in acute coronary syndromes. IFCC Scientific Division, Committee on Standardization of Markers of Cardiac Damage. International Federation of Clinical Chemistry. AB - This paper presents evidence and suggestions from the IFCC Committee on "Standardization of Markers of Cardiac Damage" (C-SMCD) on the use of biochemical markers for the triage diagnosis of acute coronary syndromes. There is general agreement that both 'early' and 'definitive' biochemical markers of myocardial damage are necessary and that these assays must be available with a turnaround time of 1 h or less. Currently, myoglobin is the marker that most effectively fits the role as an 'early' marker, whereas 'definitive' markers are cardiac troponins. Since the sensitivity of the initial electrocardiogram is only 50% for detecting myocardial infarction, the use of biochemical markers may significantly contribute to the early diagnosis and become relevant when the electrocardiogram is not diagnostic. In addition, new sensitive biochemical markers, particularly the cardiac troponins, are presently the best to detect the presence of minor myocardial cell damage. With regard to this, two decision limits are probably needed for the optimal use of troponins: a low abnormal value suggesting the presence of myocardial damage and a higher value suggesting the diagnosis of myocardial infarction according to traditionally used criteria. Properly designed studies should be performed to establish limits for each commercially available troponin assay. Finally, it is recognized that there is no need for the use of any biochemical marker when the clinical diagnosis is unequivocal, other than for diagnosing reinfarction, estimating the infarct size, and monitoring thrombolytic therapy. PMID- 10475080 TI - A new Clethrionomys-derived hantavirus from Germany: evidence for distinct genetic sublineages of Puumala viruses in Western Europe. AB - Puumala (PUU) viruses are the predominant etiologic agents of hantavirus infections in Europe. The most important reservoir is the bank vole, Clethrionomys glareolus (Cg), belonging to the subfamily Arvicolinae of the Muridae family. Here we report on the molecular characterization of the first rodent-derived sequence (PUU/Cg-Erft) from Germany. Comparison of the S and M segment coding regions revealed 92.5 and 92.8% identity, respectively, with PUU/H 9013, a human isolate from France. However, only 83.1% identity was found with the S segment of a previously reported PUU sequence from a German HFRS case (PUU/H-Berkel) indicating the co-existence of two distinct sublineages in Germany. Phylogenetic and alignment analyses of S and M segment coding regions enabled us to assign PUU viruses/sequences to at least six distinct genetic sublineages. Membership was defined by nucleotide sequence differences of < 8%, whereas a diversity of > 14% clearly outgrouped a virus/sequence. Based on S segment sequences the sublineage represented by Clethrionomys rufocanus-derived viruses from Japan diverged at a well supported node from the clade harbouring all Clethrionomys glareolus-derived European PUU viruses. A correlation between genetic relationship and geographic origin of PUU viruses was observed which may support a co-evolution of PUU viruses with distinct subspecies of their reservoir host. PMID- 10475081 TI - A persistent variant of influenza C virus fails to interact with actin filaments during viral assembly. AB - C/AA-pi virus, a variant of influenza C/Ann Arbor/1/50 virus, establishes persistent infections in MDCK cells, characterized by low levels of progeny production. During viral assembly, nucleoprotein (NP) was found homogeneously distributed over cytoplasmic and nuclear compartments and matrix (M) protein was likewise localized in a barely structured fashion. In contrast, infections with nonpersistent influenza A, B and C viruses produced cytoplasmic granular structures, which typically consisted of colocalized NP and M proteins. Studies on the in vitro interaction between NP and M proteins revealed identical binding capacities comparing influenza C wild-type virus with the persistent variant. Cytochalasin D treatment of infected cells demonstrated that NP protein of the wild-type virus, but not of the persistent variant, was distinctly associated with cellular actin filaments. Moreover, the assembly characteristics of wild type virus were modulated in the presence of recombinant persistent-type NP protein towards a behaviour similar to persistent infection. Cell type specificity was particularly illustrated in C/AA-pi virus-infected Vero cells, which did not support viral persistence, but produced granular wild-type-like complexes. Thus, interaction between NP, M and actin proteins (i) is a basic part of the viral assembly process, (ii) is dominantly modulated by NP protein and (iii) is specifically altered in the case of persistent infection. PMID- 10475082 TI - Conversion of US3-encoded protein kinase gene from pseudorabies virus in a diploid gene located within inverted repeats by genetic recombination between the viral genome isomers. AB - The pseudorabies virus (PRV) genome consists of two components, long (U(L)) and short (U(S)) regions. The U(S) region is the only one capable of inverting itself relative to the U(L) region during productive infection, generating two equimolecular isomeric forms of viral DNA. Here we describe a recombinant virus (gIp2) generated by genetic recombination between pseudorabies viral isomers. This recombination event was observed in the parental virus gIS8, which was obtained by insertion of the alpha4-TK herpes simplex virus type 1 (HSV1) gene. The growth of gIS8 virus in the presence of 5-bromodeoxyuridine (BrdU) yielded gIp2. This was generated by nonhomologous recombination either between the two viral genomic isomers of gIS8, P and I(U/S), or between the same P isomer using nonhomologous and homologous recombination, with loss of the HSV1 sequences and duplication of the PRV US3-encoded protein kinase gene. Virus gIp2 is negative for TK, gI, gE, 11K and 28K and shows an in vitro replication capacity in neuronal cells approximately 22 times lower than that of parental virus gIS8, and similar to that of the Bartha vaccine virus strain in monkey kidney and human neuronal cells. PMID- 10475083 TI - Sequence comparisons of medium RNA segment among 15 California serogroup viruses. AB - The complete nucleotide sequences have been determined for the M segment of 12 California (CAL) serogroup bunyaviruses. A method is described here of long reverse transcription-polymerase chain reaction (RT-PCR) that yields the full length medium (M) RNA genomic segment. A phylogenetic tree was constructed by comparison of the open reading frames (ORFs) in the M RNA segment of 15 CAL serogroup viruses. Three distinct branches were identified and they are represented by the California encephalitis (CE), Melao (MEL), and Trivittatus (TVT) complexes. These groups correspond to those previously established by small (S) RNA genomic sequences. In addition, except for Inkoo virus, the predicted relationship among these viruses agreed with those found by serology. PMID- 10475084 TI - The S gene of canine coronavirus, strain UCD-1, is more closely related to the S gene of transmissible gastroenteritis virus than to that of feline infectious peritonitis virus. AB - To gain insight into the genetic relationships among six canine coronavirus (CCV) strains, the variable region of the spike (S) protein gene was sequenced. The CCV strains were: two ATCC reference strains, the Insavc-1 vaccine strain, the National Veterinary Services Laboratories (Ames, IA) challenge strain, and two California field isolates (UCD-1 and UCD-2) from the 1970s. All six strains, downstream of the nucleocapsid (N) protein gene, had sufficient size for an ORF 7b, and thus, none were transmissible gastroenteritis virus (TGEV)-like since TGEV lacks ORF 7b. By sequence analysis of the variable domain at the 5' end of the S gene, five of the six CCV strains had a high degree of identity with feline infectious peritonitis virus (FIPV). However, one CCV field isolate (UCD-1) was different and had a high degree of identity with the 5' end of the TGEV S gene. This suggests that RNA recombination occurred at this site between antigenically related coronaviruses. The low passage field isolates, UCD-1 and UCD-2, varied in their initial infectivity for swine testicular cells suggesting that sequence differences in the variable domain of the S gene may account for biological variation among CCVs. PMID- 10475085 TI - Sequence comparison of the L2 and S10 genes of bluetongue viruses from the United States and the People's Republic of China. AB - Bluetongue virus (BTV) infection of ruminants is endemic throughout much of the US and China. The S10 and a portion of the L2 gene segments of Chinese prototype strains of BTV serotypes 1, 2, 3, 4, 12, 15, and 16 were sequenced and compared to the same genes of prototype and field strains of BTV from the US. Phylogenetic analysis of the S10 gene segregated the Chinese viruses into a monophyletic group distinct from the US viruses, whereas similar analysis of the L2 gene segregated strains of BTV according to serotype, regardless of geographic origin. PMID- 10475086 TI - Analysis of the intergenic region of tomato spotted wilt Tospovirus medium RNA segment. AB - The intergenic region (IGR) of the medium (M) RNA of tomato spotted wilt Tospovirus (TSWV) isolates naturally infecting peanut (groundnut), pepper, potato, stokesia, tobacco and watermelon in Georgia (GA) and a peanut isolate from Florida (FL) was cloned and sequenced. The IGR sequences were compared with one another and with respective M RNA IGRs of TSWV isolates from Brazil and Japan and other tospoviruses. The length of M IGR of GA and FL isolates varied from 271 to 277 nucleotides. The M IGRs of TSWV from potato and stokesia, and tobacco and watermelon were identical with each other in their length and sequence. IGR sequences were more conserved (95-100%) among the populations of TSWV from GA and FL, than when compared with those of TSWV isolates from other countries (83-94%). The conserved motif (CAAACTTTGG) present in the IGRs of both M and small (S) RNAs of a Brazilian isolate of TSWV was also conserved in the isolates studied. Cluster analysis of the IGR sequences showed that all GA and FL isolates are closely clustered and are distinct from the TSWV isolates from other countries as well as from other tospoviruses. PMID- 10475087 TI - Bovine papillomaviral gene expression in equine sarcoid tumours. AB - The sarcoid is a benign locally invasive dermal fibroblastic lesion, commonly affecting horses and donkeys. The aetiology of the equine sarcoid is equivocal. Bovine papillomaviral (BPV) DNA (type 1/2) is frequently demonstrable in equine sarcoid tumour biopsies. However, the exact role of the virus in the disease process and its contribution to the phenotypic differences in sarcoids is not known. It was sought to assess the transcriptional activity of BPV-1 found in sarcoid tissues. Of 20 tumours examined, 18 were positive for E2 expression and ten positive for L1 expression. Viral oncogenes E5, E6 and E7 transcripts were detected in 16, nine and 12 tumours, respectively. This study demonstrates BPV gene expression in equine sarcoids and provide the first evidence for a direct involvement of the virus in the pathogenesis of sarcoids. PMID- 10475088 TI - Characterization of the immunodominant cross-reacting epitope of visna maedi virus and caprine arthritis-encephalitis virus capsid antigen. AB - Maedi visna (MV) and caprine arthritis-encephalitis (CAE) are two retroviral infections distributed world wide. Antigenic cross reactions between the viruses have been demonstrated in gag and env encoded structural proteins. Antigens from ovine lentiviruses are easier to produce in cell culture systems and therefore have been used in the development of diagnostic tests for both infections. Antigenically relevant epitopes have been characterised in the transmembrane protein, but little information is available on the immunodominant and cross reacting epitopes in the major capsid antigen (p25). In this study four different recombinant subunits of ovine lentivirus p25 were tested against sera from infected goats and a detailed characterisation of the immunodominant subunit was carried out. Highest ELISA absorbances were obtained with a 29 amino acid subunit located in the N'-terminal half of p25. Through the analysis of overlapping peptides spanning this region we identified a 17 amino acid sequence that can be used in the development of a highly standardized synthetic peptide-based assay. PMID- 10475089 TI - Improvement in functions of the central nervous system by estrogen replacement therapy might be related with an increased nitric oxide production. AB - Estrogen promotes neurons growth, prevents neuronal cell atrophy and regulates synaptic plasticity. Administration of estrogen protects neurons against oxidative stress, excitotoxins, and beta-amyloid-induced toxicity in cell culture. It has been shown that estrogen treatment reduces the serum monoamino oxidase levels and might regulate learning and memory. Nitric oxide (NO) is a retrograde messenger and long-term potentiation can be block using NO-synthase inhibitors or can be prevent with NO-scavengers. NO synthase is widespread in the central nervous system and acts as neurotransmitter/neuromodulator. The actions of serotonin, bradykinin, endothelin, acetylcholine and noradrenaline might be linked to NO formation. Estrogen induces activity of constitutive NO synthase and estrogen replacement therapy in postmenopausal women increases significantly circulating nitrite plus nitrate levels. The effect of estrogen on NO synthesis is rapid and is maintained with repeated administration. We demonstrated the effects of estrogen replacement therapy in Andean postmenopausal women were associated with a significantly increase in plasma levels of nitrite plus nitrate. Our hypothesis is that beneficial effect of estrogen replacement therapy on involutive depression in postmenopausal women is mediated by increase in NO production by central nervous system. PMID- 10475090 TI - Clinical performance of vascular grafts lined with endothelial cells. AB - The replacement of arteries with purely synthetic vascular prostheses often leads to the failure of such reconstructions when small-diameter or low-flow locations are concerned, due in part to the thrombogenicity of the internal graft surface. In order to improve long-term patency of these grafts, the concept of endothelial cell seeding has been suggested because this metabolically active endothelial surface plays major roles in preventing in vivo blood thrombosis and because vascular grafts placed in humans do not spontaneously form an endothelial monolayer whereas they do in animal models. The composite structure resulting from the combination of biologically active cells to prosthetic materials thus creates more biocompatible vascular substitutes. To achieve endothelialization of synthetic vascular grafts, previous efforts aimed at "one-stage" procedure (adding autologous endothelial cells to the graft at the time of implantation) in the 1980's seemed clinically feasible but results of reported clinical trials were controversial and mostly disappointing. An alternative method is an in vitro complete and preformed endothelial lining at the time of implantation: the "two stage" procedure which implies harvest and culture of autologous endothelial cells. Up to date, the latter approach demonstrated its superiority in terms of significantly increased patency of the grafts that underwent endothelialization eight years earlier. PMID- 10475091 TI - The vitronectin receptor plays a role in the adhesion of human cytotrophoblast cells to endothelial cells. AB - During placental development in higher primates trophoblast cells invade maternal blood vessels and migrate along the luminal surface of endothelium. In the present study, the adherence of human cytotrophoblast cells to endothelial cells has been characterized to test the hypothesis that vitronectin receptors (alpha(v) integrins) play a role in intra-luminal trophoblast migration. Adherence was measured using a quantitative fluorescence-based assay and was found to increase in a time-dependent fashion up to about 2 h after which it leveled off. Adhesion was detectable at 4 degrees C but was greatly reduced compared to that seen at 37 degrees C. Adhesion was partially blocked by antibodies against alpha(v)beta3/beta5 integrin, beta1 integrin and by antibodies against P-selectin. Antibodies against beta3 integrin subunits had no effect. Adhesion was reduced by galactose-6-phosphate and fructose-6-phosphate. Flow cytometric analysis revealed alpha(v) integrin on the surface of cytotrophoblast and endothelial cells. Beta1 integrin was detected on the surface of endothelial cells and on cytokine-stimulated cytotrophoblast cells. Beta3 and beta5 integrins were not detected on the surface of either cell type, although beta3 was detected using permeabilized endothelial cells. These results raise the possibility that alpha(v) integrins expressed by both cytotrophoblast cells and endothelial cells, and P-selectin expressed by endothelial cells, may be important in facilitating trophoblast adhesion and migration along the uterine microvasculature. PMID- 10475092 TI - TGF-beta downmodulates cytokine-induced monocyte chemoattractant protein (MCP)-1 expression in human endothelial cells. A putative role for TGF-beta in the modulation of TNF receptor expression. AB - The expression of chemokines, including monocyte chemoattractant protein (MCP)-1, by many cell types contributes to the pathogenesis of inflammatory diseases. We examined MCP-1 expression in human umbilical vein endothelial cells (EC) following cytokine treatment. We specifically compared the effect of TGF-beta 1 on this cytokine-induced expression, as TGF-beta has been shown to have immunosuppressive effects on EC. EC expressed MCP-1 mRNA and protein in response to TNF alpha, IFN gamma or IL-1beta, but not TGF-beta1. TGF-beta1 in cotreatment with either TNF alpha or IL-1beta, but not IFN gamma, significantly decreased MCP 1 mRNA and protein expression, as compared to TNF alpha or IL-1beta treatment alone. Pretreatment with TGF-beta had no effect on any cytokine-induced MCP-1 expression. TGF-beta had no effect on MCP-mRNA stability. Examination of TNF receptor expression by flow cytometry revealed that TNF alpha treatment caused a decrease of p75 expression on the cell surface. The p55 receptor was not detected at the cell surface, but was localized intracellularly by confocal microscopy. Treatment of EC with TGF-beta alone decreased p75 surface expression and in cotreatment with TNF alpha, caused an additive decrease in p75 surface expression, as compared to TNF alpha treatment alone. Whereas mRNA expression for both receptors was increased with TNF alpha treatment, this was decreased with TGF-beta/TNF alpha cotreatment, as compared to TNF alpha treatment alone. Thus, the expression of TNF receptors was also down-modulated by TGF-beta. These findings indicate additional mechanisms by which TGF-beta exerts immunosuppressive properties on EC. PMID- 10475093 TI - Endothelin-1, endothelin receptors and ecNOS gene transcription in vital organs during traumatic shock in rats. AB - Endothelin-1 (ET-1) is a vasoconstrictor peptide that may play an important role in the pathophysiology of severe trauma. We examined ET-1 gene expression in vital organs (i.e., heart, lungs, kidneys, liver and small intestine) during murine traumatic shock using ribonuclease protection assays. Our data show that ET-1 mRNA was significantly increased in the lungs two hours after trauma when compared with control anesthetized rats. There was also a significant increase in ET-1 transcripts occurring in the kidneys, heart and liver. During these experimental conditions, we also observed statistically significant increased endothelin type B (ET(B)) receptor mRNA expression in the lung, heart, liver, kidney and small intestine. Expression of endothelial constitutive nitric oxide synthase (ecNOS) gene, which is functionally coupled to ET(B) receptor, also was increased in vital organs during traumatic shock. Endothelin type A (ET(A)) receptor gene expression was slightly decreased in the lung, liver and small intestine. These results suggest that ET-1 and ET(B) mRNA expression are mainly increased in the lung and other vital organs and may play a functional role in the pathophysiology of murine traumatic shock. PMID- 10475094 TI - High glucose induces enhanced monocyte adhesion to valvular endothelial cells via a mechanism involving ICAM-1, VCAM-1 and CD18. AB - Upon induction of experimental hyperglycemia (i.e. diabetes) pathological modifications are early detected (approximately 7 days) at the level of the cardiac valves leading rapidly to the development of valvular atheroma. Monocyte adhesion to the vascular endothelium is one of the initial event at the onset of atherosclerosis. We questioned whether high glucose enhances monocyte adhesion to the valvular endothelial cells (VEC) so as to explain, in part, the accelerated atheroma formation that occur in diabetic conditions. To this purpose we compared the adhesion of monocytes to VEC cultured in 5.5 mM (normal) glucose (NG) or in 33 mM (high) glucose (HG) or in high mannitol (HM) (27.5 mM mannitol plus 5.5 mM glucose), a concentration known to simulate the hyperosmolar effect of high glucose. After incubation for 30 min at 37 degrees C, the adhesion of monocyte cell line (U937 cells) to VEC was quantitated by a fluorimetric assay or by direct counting. Statistical data showed a significant increased adhesion of monocytes to VEC grown in HG (up to 4 fold) or in HM (up to 2.7) when compared to normal conditions. Using a battery of specific monoclonal antibodies molecules it was found that the increased adhesion of monocytes to VEC grown in high glucose was specifically inhibited (p < 0.05) by anti-ICAM-1, anti-VCAM-1 and anti-CD18 monoclonal antibodies. Together, the results indicate that high glucose induces enhanced monocyte adhesion to VEC via a mechanism involving in part an osmotic effect and mainly the cell adhesion molecules: ICAM-1, VCAM-1 and CD18. PMID- 10475095 TI - Apoptosis induced by lack of hemodynamic forces is a general endothelial feature even occuring in immortalized cell lines. AB - Confluent monolayers of primary endothelial cells display a high viability and an apparently constant cell density. However upon prolonged cultivation the monolayer degenerates with increasing numbers of senescent cells finally representing the whole culture. Recently we showed that lack of hemodynamic forces induces apoptosis in organ cultures as well as in confluent monolayers of human umbilical cord vein endothelial cells (HUVEC). The apoptosis started at a low level and was counteracted by a continuous proliferation of the remaining cells. Here we show that the induction of apoptosis by lack of hemodynamic forces is a general characteristic of vascular endothelial cells, valid for endothelial cells from various organs and species: human umbilical cord vein endothelial cells (HUVEC), human microvascular placental endothelial cells (HPEC) and bovine aorta endothelial cells (BAEC). Furthermore apoptosis due to the lack of hemodynamic forces can also be induced in various endothelial cell lines: EA.hy 926 derived from HUVEC and PBMEC-A1 derived from PBMEC. However degeneration of confluent monolayers does not occur with these cell lines even in monolayers kept for several weeks. This indicates that the degeneration of normal endothelial cell monolayers is caused by depletion of the proliferation potential of the endothelial cells. PMID- 10475096 TI - Apoptosis inducers endotoxin and Fas-ligation enhance the expression of vascular endothelial growth factor in human endothelial cells. AB - Endothelial cells in culture were exposed during four hours to the apoptosis inducing agents endotoxin (lipopolysaccharide, LPS) and Fas-ligand mimicking antibody in various concentrations. With addition of a deletion primer as internal standard a competitive RT-PCR was performed to measure semi quantitatively the expression of mRNA of Vascular endothelial growth factor (VEGF). It appeared that endothelial cells survive increasing amounts of LPS and show a concentration- and time-dependent increase in the expression of VEGF-mRNA. The same effect was found with Fas-ligation, although at high concentrations Fas ligation induced no further increase, but even a decrease of VEGF expression, possibly related to cell damage. Apoptotic cells were rarely observed after LPS stimulation, but simultaneous incubation with a blocking antibody to VEGF resulted in a significant increase in apoptosis. We hypothesize that endothelial cells are resistant to apoptosis induction by autocrine expression of VEGF under stress conditions. PMID- 10475097 TI - Official satellite symposium of the forum meeting of the European Neuroscience Association. Signal transduction pathways in the blood-brain barrier. Hotel Intercontinental, Berlin, Germany. PMID- 10475098 TI - Heightened IgE response to mite antigens in inflammatory neuropathies. AB - In order to clarify the IgE response to common environmental antigens, we measured the serum total IgE and allergen-specific IgE in 50 patients with Guillain-Barre syndrome (GBS), nine patients with Fisher syndrome (FS), 14 patients with chronic inflammatory demyelinating polyneuropathy (CIDP), 18 patients with mononeuritis multiplex (MNM), 43 patients with neurodegenerative disorders and 82 healthy controls by ELISA. The total IgE level was significantly higher in patients with GBS (median = 135 U/ml, P<0.05), CIDP (median = 175 U/ml, P<0.05) and MNM (median= 199 U/ml, P<0.05), than in the healthy controls (median = 79 U/ml), but not in those patients with neurodegenerative disorders. The specific IgE to Dermatophagoides pteronyssinus was significantly higher in the patients with GBS (56%, P<0.01) and MNM (72%, P<0.005) than in the healthy controls (32%). The level of specific IgE to Dermatophagoides farinae tended to be higher in the patients with GBS than in the healthy controls (0.05 or =10 mm. This is a report of a 54-year-old woman with endometrial cancer who presented with nausea, vomiting and abdominal pain 1 week following laparoscopically-assisted vaginal hysterectomy, bilateral salpingo-oophorectomy, and lymphadenectomy. Abdominal radiographs and computed tomography demonstrated small bowel obstruction and herniation through a 5-mm trocar site. Reduction of the hernia and closure of the fascial incision were performed at exploratory laparotomy with normal recovery. Bowel herniation can occur through 5-mm trocar sites following prolonged operative laparoscopy. The peritoneum and fascia of these incisions should be closed. PMID- 10475122 TI - Primary intravenous paclitaxel and platinum chemotherapy for high-risk Stage I epithelial ovarian carcinoma. AB - OBJECTIVE: To evaluate the efficacy of intravenous (i.v.) paclitaxel and platinum chemotherapy in patients with high-risk Stage I epithelial ovarian carcinoma. METHODS: We performed a retrospective chart review of all patients with Stage I ovarian cancer treated at our institution between March 1993 and June 1995. RESULTS: Twenty patients received adjuvant paclitaxel-containing chemotherapy for Stage I ovarian carcinoma after comprehensive surgical staging. Five patients (25%) had Stage IA disease and 15 patients (75%) had Stage IC disease. Tumor grades were: 1, five patients (25%); 2, nine patients (45%); and 3, six patients (30%). Histologic cell types were: clear-cell, ten (50%); endometrioid, five (25%); mucinous, three (15%); and serous, two (10%). Nineteen patients (95%) were treated with i.v. paclitaxel and platinum chemotherapy. One patient (5%) received i.v. paclitaxel alone. Eighteen patients (90%) had five cycles of chemotherapy, while two patients (10%) had three. The 96 total cycles were associated with nine episodes (9%) of significant toxicity: fever, four (4%); severe nausea and vomiting, two (2%); Clostridium difficile enteritis, one (1%); congestive heart failure, one (1%); and anemia, requiring blood transfusion, one (1%). With a median follow-up of 36 months (range 24-50 mos), all 20 patients are alive, and 19 (95%) are disease-free. The one patient (5%) treated with i.v.++paclitaxel alone developed an abdominal recurrence 22 months after diagnosis. CONCLUSION: Primary i.v.++paclitaxel and platinum chemotherapy in patients with high-risk Stage I epithelial ovarian carcinoma is reasonably well tolerated and may improve survival. Larger studies with long-term follow-up are needed. PMID- 10475123 TI - A rare case of solitary metastasis of cervical cancer to the colon after radiation therapy. PMID- 10475124 TI - Paget's disease of mammary skin without nipple involvement. A report of three cases. PMID- 10475125 TI - Benign and malignant mammary tumors induced by DMBA in female Wistar rats. AB - This study pretends to characterize 7, 12-dimetylbenz[a]anthracene-induced benign and malignant tumors. One hundred and twenty female Wistar rats were randomly allocated to two groups: Control Group and Induction Group; IG animals were given a single dose of DMBA and killed 24 weeks after. Other tumors besides breast tumors were diagnosed, mainly tumors of the salivary glands and ovarian benign epithelial tumors. Incidence of breast disorders was about 60%. Macroscopic mammary tumors varied in dimension from 2 mm to 55 mm. Malignant breast tumors (n = 56) were essentially invasive ductal carcinomas (91.1%), G1 (92.2%), presenting histologic characteristics of good prognosis. Predominant benign breast disorders consisted of glandular (68.6%) and atypical (20%) hyperplasias reproducing histologic types of human breast diseases. Different individual susceptibility to DMBA apparently occurs; while some rats never developed neoplasias, others exhibited several tumors. PMID- 10475126 TI - Characterization of a MAb to ovarian cancer and its possible diagnostic application. AB - We report the distribution pattern of the target antigen of an IgG1 monoclonal antibody, Ki-OC III raised in BALB/C mice against solubilised ovarian adenocarcinoma, in normal tissue and in a collection of human tumour types. Special reference was made to benign and malignant ovarian tumours. The reactive antigen protein purified to homogeneity for a planned amino acid analysis showed three bands between 37-40 kDa. Immunohistochemical analysis revealed a specificity of 98% and a sensitivity of 77%. The tracer kinetics of the radiolabelled antibody were tested on a human ovarian carcinoma cell line in athymic mice. The results were compared to cell lines derived from breast and stomach carcinomas as well as to a human glioblastoma cell line. The results show the preferential uptake by ovarian cancer cells followed by breast cancer cell lines. In animal models, scintigraphic monitoring and direct measurement of the radio-labelled monoclonal antibody showed a preferential accumulation in tumours, in addition to high level signals in the liver, kidney, spleen and heart which were related to degradation, excretion and high circulation. The Ki-OC III reactive antigen could be a potential candidate for immunomonitoring of ovarian and possibly also of breast cancer, for in vivo tumour imaging as well as for histopathologic examinations. PMID- 10475127 TI - Estimation of the usefulness of neoplastic markers TPS and CA 125 in diagnosis and monitoring of ovarian cancer. AB - PURPOSE: We have undertaken an attempt to compare the suitability of tumor markers TPS (tissue polypeptide specific antigen) and CA125 for diagnosis and monitoring of ovarian cancer patients. METHODS AND MATERIAL: The studies were performed on 33 patients treated for ovarian cancer in the Department of Oncology, Karol Marcinkowski School of Medicine, Poznan from 1995-1996, Serum levels of TPS and CA125 were determined before surgery and at each chemotherapy course. CONCLUSION: Estimation of the neoplastic markers TPS and CA125 is suitable for diagnosis of ovarian cancer. Parallel use of TPS and CA125 in ovarian cancer patients increases sensitivity of the diagnosis. Estimation of TPS is highly suitable and estimation of CA125 is of low value in detection of mucinous ovarian cancer. Serum levels of the neoplastic markers TPS and CA125 decrease after total or debulking surgery for ovarian cancer. Serum TPS and CA125 levels reflect the course of the neoplastic process during chemotherapy. PMID- 10475128 TI - Epidemiology of preinvasive lesions. AB - OBJECTIVE: Review of epidemiological data on pre-invasive cervical lesions. MATERIAL AND METHODS: Literature review and analysis of data from our Department. RESULTS: Prevalence of data on preinvasive cervical lesions varies widely and depends on factors such as differences among countries or regions and among ethnic groups, and especially, differences in the type of population studied. Most important risk factors are: number of sexual partners, smoking, contraceptive use, HPV, age at first intercourse, and screening. CONCLUSIONS: In order to reduce risk, pap smears should be performed regularly, safe sex practices should be recommended, and the use of tobacco products should be avoided. PMID- 10475129 TI - Transplacental effects of IgG generated against soluble 53 kDa protein on the splenic lymph system of rat progeny exposed to carcinogen: rate of apoptosis, proliferation of lymphocytes and expression of Fas and Fas ligand proteins. AB - BACKGROUND: We have previously shown that vaccination with IgG generated against the soluble 53 kDa (s53) protein modified the splenic response to carcinogens. Here we studied whether such immunization could affect the splenic lymphatic system of the offspring. METHODS: Offspring of normal female rats or of rats immunized with anti-s53 IgG were exposed to a carcinogen (dimethyl benz(a)antracene). After 4 months, their spleens were resected and evaluated immunohistochemically for lymphocyte proliferation, apoptosis and apoptosis related proteins (Fas and Fas ligand), in tumor-free and tumor-bearing animals. RESULTS: Spleens of progeny of unvaccinated rats had a significant decrease in the areas of follicles, germinal centers and the mantle layer after exposure to carcinogens, while maternal vaccination resulted in a significant expansion of the progeny's splenic follicles and germinal centers, the zones of B cell proliferation. The area of periarterial lymph sheaths (PALS) expanded in these offspring, reflecting activation of the T-zone. Maternal vaccination also resulted in a significant rise of Fas ligand-positive lymphocytes in the follicles and PALS of their tumor-free offspring. Tumorigenesis stimulated the Fas activity of B and T cells in the spleens, and this was much enhanced by maternal vaccination. CONCLUSIONS: Maternal vaccination before pregnancy results in altered morphological and functional attributes of the splenic immune system of the offspring. This increased immunoreactivity could reduce the risk of tumors in progeny of vaccinated mothers. PMID- 10475130 TI - Analysis of p53 and K-ras genes and their proteins in a sarcoma botryoides of the uterine cervix. AB - Several data indicate that the activation of oncogenes and growth factors as well as inactivation of the tumor suppressor genes are implicated in the development of human neoplasms, including sarcomas. In the present study we described a case of the extremely rare, but highly malignant neoplasm of the female genital tract known as sarcoma botryoides of the uterine cervix and assessed, using molecular and an immunohistochemical analysis, p53 and K-ras alterations in the tumor. A point mutation in exon 6 of the p53 tumor suppressor gene was found but no K-ras gene point mutations at codons 12, 13 and 61 were detected using molecular analysis. p53 protein was overexpressed in more than half of the neoplastic cells, however, ras p21 protein expression was not immunohistochemically detected. Our data indicate that p53, but not K-ras gene alterations may play a role in the development and progression of sarcoma botryoides of the uterine cervix. PMID- 10475131 TI - The value of serum CA 125 and association CA 125/CA 19-9 in endometrial carcinoma. AB - One hundred and twelve women with endometrial carcinoma were studied with serum sampling to determine preoperative and postoperative levels of CEA, CA 15-3, CA 19-9, TPA and CA 125. After surgical treatment 88 patients had stage I, 8 stage II, 14 stage III and 2 stage IV disease. Before treatment the sensitivity of CEA, CA 15-3, CA 19-9, TPA and CA 125 was 22.3% (25/112), 32.1% (36/112), 22.3% (25/112), 45.5% (51/112), 33.9% (38/112), respectively. According to pathological stage a statistically significant difference between intrauterine (96 cases) and extrauterine disease (16 cases) was noted only for CA 125 (28.1% vs. 68.7%) and CA 15.3 (28.1% vs. 56.2%). In relation to histological grading CA 125 rises progressively from well-differentiated cases to poorly-differentiated tumors. During the follow-up the most reliable marker was CA 125: values more than 35 U/ml of this marker resulted positive in 50% of relapsed cases and only in 5.1% of disease-free cases, thus demonstrating a high specificity. The association of various markers during the follow-up allowed us to reveal interesting results only for the CA 125/CA 19-9 combination. In fact the combined use of these markers permitted a high sensitivity (83.3%), with only 12.8% false positive cases, so with a high specificity. PMID- 10475132 TI - An unusual case of malignant Brenner tumor in association with low-grade urothelial carcinoma of the urinary bladder. A case report. AB - We present a case of a 45-year-old Caucasian female with a malignant Brenner tumor arising at the site of the right ovary 21 years after right adnexectomy performed for ectopic pregnancy. Several months after the presentation of the malignant Brenner tumor the patient was diagnosed with low-grade urothelial carcinoma of the urinary bladder. All clinical, biochemical, and pathological findings with the emphasis on the differential diagnosis are summarized. A brief survey of histological theories and classification of Brenner tumors as well as therapeutic approaches with their results are given. We have found a few papers [1, 2] dealing with the coexistence of Brenner tumors with urothelial ones, but to our knowledge the coexistence of both malignant tumors has not been published previously. PMID- 10475133 TI - Anogenital intraepithelial lesions in HIV positive patients. Report of 3 cases with 3-year follow-up. AB - Three cases of HIV-positive immunocompromised women, complicated by anogenital intraepithelial lesions are presented in the present paper. Two patients, aged 42 and 33, had a combination of cervical intraepithelial neoplasia grade 3 (CIN 3) and anal intraepithelial neoplasia grade 2 (AIN 2). The other one, aged 26, presented an association of CIN 2 and AIN 1. All lesions were HPV-associated and proved by colposcopically-guided biopsies. PMID- 10475134 TI - Acquired protein C deficiency following cisplatinum-navelbine administration for locally advanced breast cancer. Case report. AB - Thromboembolic events have recently been reported following diverse regimens of chemotherapy for breast cancer. This is a report of a 39-year-old woman, a diagnosed case of locally advanced breast cancer, who received many regimens of chemotherapy. She presented with deep venous thrombosis 2 months after starting the cisplatinum-navelbine regimen. Protein C deficiency was the only abnormal coagulation test that normalized after cessation of chemotherapy. PMID- 10475135 TI - Benign cystic mesothelioma of the peritoneum an immunopathological study of three cases. PMID- 10475136 TI - Uterine leiomyosarcoma in a postmenopausal woman treated with tamoxifen: case report. PMID- 10475137 TI - Intraoperative assessment of depth of myometrial invasion in endometrial carcinoma. AB - OBJECTIVE: To evaluate the adequacy of intraoperative assessment of depth of myometrial invasion in patients with endometrial adenocarcinoma. METHODS: Of the 58 evaluable cases, depth of myometrial invasion was estimated by gross examination of fresh tissue by an experienced surgeon and a pathologist and on the frozen section by the same pathologist. This was compared with the depth of invasion on the final microscopic examination performed by another pathologist. RESULTS: The depth of invasion was accurately predicted by the surgeon in 89.7% of the patients, while the pathologist's accurate prediction rates on fresh tissue and frozen section were 86.2% and 91.4%, respectively. The accurate prediction rate gradually diminished for both the surgeon and the pathologist as the histologic grade increased. Frozen section examination was reliable in grade I cancer (100%), while gross examination of the surgeon and the pathologist had a significant error rate in predicting accurate depth of invasion (7.6%-33%). CONCLUSION: If frozen section shows that myometrial invasion in patients with grade 1 endometrial carcinoma is less than 1/3, lymphadenectomy may be omitted. In all other cases radical surgery and surgical staging is mandatory to avoid undertreatment. PMID- 10475138 TI - Minimal injury mastectomy in the surgical treatment of stage I and II breast cancer patients. A 20-year experience. AB - The main purpose of the present study is to enhance what we coined minimal injury mastectomy (MIM) as a reliable, efficient and acceptable procedure in the surgical treatment of breast cancer besides other therapeutic options currently used in the setting of adjuvant treatment (e.g. radiation therapy, chemotherapy or hormonal manipulations). We analysed retrospectively data of 142 patients with stage I and II breast cancer. Fifty-five of them had stage I, and the remaining 87 patients had stage II disease. Patients were submitted to minimal injury mastectomy, which is a broad segmental mastectomy, with limited or no axillary lymphadenectomy, thus minimizing locoregional trauma. Radiotherapy was given on a selective basis. Postoperative follow-up period ranged from one to 19.5 years (mean 103.5 months). Cosmetic results, shoulder function, texture of the breast and patient psychology were excellent in all the patients. In 55 stage I patients, six local recurrences occurred (12.72%), a mean 49.9 months after MIM, with ultimate local control of all patients and a remarkable survival after treatment of recurrences. In 87 stage II patients, six local (6.8%) and 16 systemic recurrences (18.7%) occurred, the latter after a mean period of 54.4 months postoperatively. It is concluded that with the minimal injury mastectomy, locoregional injury is minimized, preserving the regional immunological functions and by preventing lymphedema of the remaining breast, systemic resistance mechanisms are maintained and other adverse factors are avoided. PMID- 10475139 TI - Mechanism-based PK/PD model for the lymphocytopenia induced by endogenous and exogenous corticosteroids. AB - OBJECTIVE: Lymphocytopenia is a sensitive surrogate marker for the immunological effects of corticosteroids. This pharmacokinetic/pharmacodynamic (PK/PD) study investigated whether the circadian variation of blood lymphocytes observed after placebo is secondary to the circadian rhythm of endogenous cortisol, and developed based on this relationship an improved PK/PD model for a more sensitive description of the effect of low-dose corticosteroid therapy on blood lymphocytes considering the net activity of the exogenous corticosteroid budesonide and endogenous cortisol. METHODS: In an open, parallel study design, 3 mg oral budesonide or placebo were given at 8.00 a.m., 4.00 p.m. and midnight to two groups of 12 volunteers. Lymphocyte counts and serum concentrations of budesonide and cortisol were monitored for 24 hours. A mechanism-based PK/PD model which considered the non-linear protein binding of cortisol and the budesonide-induced cortisol suppression was employed to relate changes in blood lymphocytes to free cortisol levels after placebo and to the net activity of free budesonide and free endogenous cortisol after active treatment. RESULTS: The circadian rhythm of blood lymphocytes observed after placebo could inversely be related to the circadian rhythm of serum cortisol. After budesonide administration, lymphocyte counts could accurately be linked to the net activity of budesonide and endogenous cortisol. The resulting EC50 values for the effect of budesonide on cortisol, budesonide on lymphocytes and cortisol on lymphocytes were 0.063 +/- 0.034, 0.22 +/- 0.13 and 26.3 +/- 15.0 ng/ml (placebo group 15.4 +/- 3.4 ng/ml), respectively. CONCLUSIONS: The presented mechanism-based PK/PD model suggests that blood lymphocytes are under physiological control of cortisol. It further indicates that endogenous and exogenous corticosteroids and their pharmacological interaction need to be considered for modeling the effects of low doses of exogenous corticosteroids on the immune system. PMID- 10475140 TI - Comparison of tilidine/naloxone, tramadol and bromfenac in experimental pain: a double-blind randomized crossover study in healthy human volunteers. AB - AIM: The analgesic efficacy and safety of single oral doses of two centrally acting compounds, the combination of 50 mg tilidine and 4 mg naloxone (Valoron N) and 50 mg tramadol (Tramal), were compared to 25, 50 and 75 mg of the non steroidal antiinflammatory bromfenac in experimental pain. SUBJECTS AND METHODS: It was a placebo-controlled double-blind 6-way crossover study design with 12 human volunteers. Acute pain was generated by electrical tooth pulp stimulation. Treatment effects were determined by recording somatosensory-evoked potentials and by subjective pain rating. RESULTS: The tilidine/naloxone combination clearly was the most potent medication in this study, followed by bromfenac 75 mg, which produced an early pain relief. Tramadol produced poor analgesia, as did bromfenac 25 and 50 mg. There was no dose-response relationship for bromfenac. Control of plasma levels revealed pronounced interindividual differences in peak plasma concentrations for bromfenac, but not for tramadol. Tilidine/naloxone exerted adverse effects in 9, tramadol in 3 volunteers. Under medication with 25 and 50 mg bromfenac, respectively, only one subject reported adverse effects. No adverse effects were experienced with 75 mg bromfenac or placebo. CONCLUSION: The results support previous conclusions about the analgesic efficacy of tilidine/naloxone and tramadol in experimental pain. Moreover, the findings suggest that 75 mg bromfenac might be suitable for fast but short relief of pain of non-inflammatory genesis. PMID- 10475141 TI - Bioavailability and selected pharmacokinetic parameters of clindamycin hydrochloride after administration of a new 600 mg tablet formulation. AB - The study was conducted to investigate the pharmacokinetics and relative bioavailability of clindamycin after administration of two oral clindamycin HCl formulations. A new tablet preparation containing 600 mg clindamycin (Clinda-saar 600, test) was compared to a marketed capsule containing 300 mg clindamycin (Sobelin 300, reference). Both preparations revealed comparable in vitro dissolution profiles with high batch conformity and homogeneity. Twenty healthy male volunteers received single doses of 600 mg clindamycin (test: 1 tablet, reference: 2 capsules) in an open, randomized, two-period crossover design. Blood samples were drawn up to 14 h p.a. and clindamycin plasma concentrations were measured using a sensitive and specific HPLC-UV method. Pharmacokinetic characteristics were similar for both preparations, arithmetic mean values (standard deviation) were computed as: AUC(0-infinity) 12.2 (4.2) and 13.1 (4.6) microg x h/ml, Cmax 3.1 (0.8) and 3.4 (0.8) microg/ml, t(max) 0.83 (0.24) and 0.85 (0.34) h, t(1/2) 2.3 (0.4) and 2.3 (0.6) h for test and reference, respectively. Mean relative bioavailability (point estimate) was 93% for AUC and 91% for Cmax. 90% confidence intervals for AUC and Cmax were within the predefined bioequivalence acceptance limits. Bioequivalence of test and reference preparations could be demonstrated. Single doses of 600 mg clindamycin orally were well tolerated without relevant differences between both preparations. PMID- 10475142 TI - Absolute bioavailability, pharmacokinetics, renal and biliary clearance of distigmine after a single oral dose in comparison to i.v. administration of 14C distigmine-bromide in healthy volunteers. AB - AIM: The aim of the study was to determine the absolute bioavailability and pharmacokinetics after a single dose oral administration in comparison to i.v. administration of 14C-labelled distigmine-bromide (14C-Ubretid) in healthy male volunteers. RESULTS: After the intravenous administration, distigmine is eliminated from the body by renal excretion (85%), and for a small fraction by biliary excretion in the feces (4%). This situation is reversed after an oral administration, where 6.5% of the dose is recovered from the urine and 88% from the feces. This means that distigmine after oral administration is hardly absorbed, the calculated bioavailability is 4.65%. CONCLUSION: The mean absorption time (MAT) after oral administration was 10 h, influencing the t(1/2alpha) (1.4 vs 4.5 h) and the t(1/2beta) (60 vs 70 h) to higher values than after the i.v. administration (p < 0.05). PMID- 10475143 TI - Pharmacokinetic variability of nimodipine disposition after single and multiple oral dosing to hypertensive renal failure patients: parametric and nonparametric population analysis. AB - OBJECTIVES: To explore the contribution of renal failure to nimodipine overall pharmacokinetic variability after single and multiple oral dosing and to develop a population pharmacokinetic model by means of the nonparametric expectation maximization (NPEM2) algorithm based on sampled individual drug concentrations close to the estimated patients' C(SS)avs (NPEM2-C(SS)av). PATIENTS, MATERIALS AND METHODS: 24 hypertensive patients with normal and reduced renal function, without clinical and laboratory data for hepatic dysfunction, were enrolled in the study and their nimodipine plasma levels were analyzed by means of a parametric and nonparametric population pharmacokinetic modeling using a maximum a posteriori Bayesian (MAPB) estimator in an iterative two-stage Bayesian population modeling program and NPEM2-algorithm. RESULTS: Comparison of parameter dispersion revealed higher variability of nimodipine disposition after the first dose than at steady-state except for apparent volume of distribution at steady state, V(SS)/F, whose variability increased from 98% to 223%. The most variable was mean residence time, MRT, whose coefficient of variation (CV) was 288% after the first dose and decreased by more than 2 times at steady-state, followed by terminal elimination half-life, t(1/2el), with CV = 171% after the first dosing and decreasing by more than 3 times at steady-state. Concerning the impact of renal failure on disposition parameters variability, patients with slightly to moderately reduced renal function, creatinine clearances between 51 to 80 and 25 to 50 ml/min, resp., stated higher variation than patients with more definitively altered renal function. The validation of NPEM2-C(SS)av population model was performed by using a set of 272 individual plasma drug concentrations, including trough levels as well as concentrations belonging to mono-exponential elimination phases after single and multiple dosing. Bayesian forecasting, using 4 trough levels per patient as Bayesian priors, revealed highly significant correlation between observed and population model predicted drug concentrations (r = 0.526, p < 0.0001). The predictive performance of NPEM2-C(SS)av population model was characterized by low bias (mean error = -0.48 microg/l, 95% CI = -0.99-0.04 microg/l), and good precision (root mean squared error = 4.32 microg/l, 95% CI = 2.53-11.17 microg/l). CONCLUSIONS: As predicted for high hepatic clearance drugs [Rowland 1985], nimodipine parameters variability decreased after reaching steady state. NPEM2-C(SS)av population model demonstrated high accuracy and precision in predicting drug levels from terminal exponential phase including trough levels at steady-state. PMID- 10475144 TI - Pharmacokinetics and hemodynamic effects of diltiazem in healthy volunteers: comparing resting with the effect of exercise. AB - PURPOSE: To determine the steady-state plasma concentrations of diltiazem (DTZ) and hemodynamic effect in humans at rest and during exercise. METHODS: Healthy volunteers (10 F, mean age 22, and 11 M, mean age 24) were recruited. Prior to receiving DTZ, each volunteer performed two 3-minute stages of treadmill exercise according to the Bruce protocol. Intra-arterial BP and ECG recordings were obtained before, during and immediately post exercise. Each volunteer then received DTZ 60 mg qid for one week. The same exercise protocol was repeated 1 h after the last dose. Steady-state plasma concentrations of DTZ were determined by a previously reported HPLC. RESULTS: DTZ decreased resting DBP from 84 +/- 13 to 79 +/- 10 mmHg (p > 0.05), and HR from 89 +/- 11 to 82 +/- 13 bpm (p < 0.05). During exercise, an average of 32 and 10% increase in SBP and DBP, respectively, and a 47% increase of HR was found (p < 0.05). DTZ limited these increases to 21% for SBP, 5% for DBP, and 44% for HR (p < 0.05 for drug effect). Steady-state plasma DTZ concentrations were 141 +/- 56 ng/ml. CONCLUSION: DTZ significantly decreased resting HR but not BP in health volunteers. It decreased both hemodynamic variables during exercise. Thus, the hemodynamic effects of diltiazem are more profound during exercise, and may be more useful surrogate markers for calcium antagonists and other cardiovascular agents in healthy volunteer studies. PMID- 10475145 TI - Effect of the interaction of TSH and insulin on the stimulation of 2-deoxyglucose uptake in FRTL-5 cells. AB - Since thyroid glycogen stores are low, the uptake of glucose is very important in order to maintain cell function (house-keeping). Previous studies have shown that TSH and insulin, independently, are regulators of this parameter. Since their corresponding mechanisms of action are different, we investigated the possible effect of the interaction between TSH and insulin on the stimulation of 2 deoxyglucose (2-DOG) uptake, a non metabolizable derivative of glucose. Confluent FRTL-5 cells were submitted to different treatments, usually for 72 h. In one series of experiments the concentration of TSH was kept constant, at 1 U/l, and the addition of insulin, from 0.16 to 1.6 micromol/l caused a progressive synergic increase in DOG uptake. When insulin concentration was kept constant, increasing amounts of TSH, from 0.5 to 10 U/l), also caused a synergic stimulation of DOG uptake. The effect of insulin was mimicked by IGF-1 (1-10 nmol/l), while that of TSH was mimicked by forskolin. Timecourse studies showed that TSH had a peak at 3 h of incubation, while insulin caused a progressive increase for up to 72 h. At short incubation times, up to 6 h, an additive effect of TSH and insulin was observed, while at longer times the interaction was synergic. The present results suggest that the interaction between the cAMP and the tyrosine kinase pathways on DOG uptake would involve two different mechanisms. At early times the effects of both hormones are additive, while in longer periods it becomes synergic. PMID- 10475146 TI - Evaluation of goiter endemia by ultrasound in schoolchildren in Val Sarmento (Italy). AB - Ultrasonography is an excellent and objective method for assessing thyroid volume, especially in children where clinical evaluation is inaccurate. The aim of this study was to evaluate the presence of goiter by thyroid ultrasound and palpation in 244 schoolchildren, 6 to 14 years old, living in some rural villages of Val Sarmento, a mountain area of Basilicata, Italy. In 1996 we revealed the presence of endemic goiter in 25% of the schoolchildren evaluated by palpation, according to World Health Organization (WHO) criteria, and in 15.9% of the schoolchildren evaluated by ultrasonography (7.5 MHz linear probe). The median urinary iodine excretion, taken from an extemporaneous sample of the first urines in the morning, was 62.2 microg/l. This study includes Val Sarmento, an area with mild-moderate grade (Grade I) of iodine deficiency, suggesting the need for iodine prophylaxis. Furthermore, it proves that the measurement of thyroid volume by ultrasonography is an essential instrumental method for a correct epidemiological study of endemic goiter, particularly in areas where there is mild iodine deficiency. PMID- 10475147 TI - Event-related brain potentials in male hypogonadism. AB - Several studies based on psychometric tests have determined an impairment of cognitive functions in patients with androgen deficiency. However, little is known about event-related potentials (ERPs) alterations in male hypogonadism. We investigated alterations of ERP in male hypogonadism before and 3 months after gonadotropin treatment. ERPs were elicited in 20 untreated male patients with idiopathic hypogonadotropic hypogonadism (IHH) (mean age: 21.1+/-1.4 years) and in a group of 30 male controls with comparable mean age and educational level. ERP recordings were repeated 3 months after hCG/hMG treatment. Untreated hypogonadal patients had longer mean P300 latencies and increased P300 amplitudes when compared to those in controls (321.6+/-18.5 vs 299.3+/-20.1 msec, p=0.0002; 12.15+/-4.47 vs 9.38+/-3.02 microV, p=0.011, respectively). The mean P300 latencies did not change significantly 3 months after gonadotropin treatment, while P300 amplitudes were decreased significantly. P300 latencies did not correlate with serum testosterone and other hormone levels. We conclude that prolongation of P300 latencies and increased P300 amplitudes are associated with male hypogonadism, but P300 prolongation is not reversed 3 months after gonadotropin treatment. These findings confirm the occurrence of cognitive defects in hypogonadal patients and would support the hypothesis that perinatal androgen deficiency contributes to an insufficient cognitive development. PMID- 10475148 TI - Natriuretic peptides receptors in human aldosterone-secreting adenomas. AB - Atrial natriuretic peptide (ANP) and B-type or brain natriuretic peptide (BNP) inhibit aldosterone secretion in humans both in vitro and in vivo. Unresponsiveness of aldosterone-secreting adenomas (aldosteronomas) to ANP in vitro and in vivo, might be due to reduced expression of the biologically-active natriuretic peptide receptor type A (NPr-A) and/or increased expression of the clearance receptor for natriuretic peptides (NPr-C). Therefore, we have analyzed NPr gene expression and ANP binding sites in human adrenals and aldosteronomas. Using reverse transcription and polymerase chain reaction, we cloned and characterized cDNAs for NPr-A, NPr-C, and the receptor (NPr-B) for the C-type natriuretic peptide (CNP). Total RNA from three normal human adrenals (obtained at surgery from patients with renal cancer) and five aldosteronomas were used for Northern analysis. NPr-A mRNA (approximately 4 kb) and NPr-B mRNA (approximately 4 kb) were expressed without significant differences in adrenals and in aldosteronomas except in an aldosteronomas that contained only very low amounts of NPr mRNAs. The gene expression of NPr-C was barely detectable both in adrenals and in aldosteronomas. ANP binding sites were analyzed by autoradiography with 125I-labeled ligand in other six aldosteronomas. Only one of the adenomas analyzed showed ANP binding sites with density of granules similar to nonadenomatous glomerulosa, whereas the others had significantly reduced densities. In summary, aldosteronomas express the genes encoding for NPr but mainly NPr-A, similarly to control adrenals. On the contrary, the binding sites for ANP are greatly reduced in most aldosteronomas. A somatic mutation or a post transcriptional defect that reduces ANP binding sites might be present in some aldosteronomas. PMID- 10475149 TI - Derangements of pyruvate dehydrogenase in circulating lymphocytes of NIDDM patients and their healthy offspring. AB - Pyruvate dehydrogenase (PDH) is poorly active in circulating lymphocytes of NIDDM patients; in vitro, it is unresponsive to insulin at 5 microU/ml and activated at 50 microU/ml, instead of activated and inhibited as in healthy controls. This study examines whether healthy offspring of NIDDM patients with a family history for this disease have these alterations. Twenty seven healthy offspring (23+/-10 yr, median 18 yr) and their parents (13 diabetic with a family history for NIDDM and 11 healthy without this history) were enrolled. Twenty healthy individuals without the history and matched for age and gender with the offspring served as controls. Minimum levels for enzyme activity before and after cell stimulation with insulin at 5 microU/ml were computed for a 95% CI with no more than 5% of the controls excluded. Increased or unvaried enzyme activity in response to insulin at 50 microU/ml was defined as abnormal. All NIDDM parents and 11/27 offspring had below normal enzyme activity and defective and reversed enzyme response to insulin at 5 and 50 microU/ml; three offspring had altered enzyme response to insulin at both concentrations, four to insulin at 5 microU/ml, three to insulin at 50 microU/ml and six, together with the healthy parents, had no alterations. We conclude that in healthy individuals a family history for NIDDM is frequently signaled, irrespective of age, by molecular derangements, with an apparent genetic background, in their circulating lymphocytes. PMID- 10475150 TI - The role of somatostatin (octreotide) in the regulation of melatonin secretion in healthy volunteers and in patients with primary hypothyroidism. AB - Somatostatin has been found in the pineal gland of several animal species, which suggests that it may be involved in the regulation of melatonin secretion. Whether somatostatin has regulatory influence on melatonin secretion in man has never been unequivocally shown. We studied the nocturnal melatonin secretion in 8 healthy volunteers, and 6 women with untreated primary hypothyroidism, a disease state that is associated with increased nocturnal secretion of melatonin. The participants were given subcutaneous injections at 18:00 h and 23:00 h of either saline or octreotide (Sandostatin; each injection 50 microg). During the nights when the healthy volunteers were given octreotide, melatonin secretion was similar to that recorded during administration of saline. Also the urinary excretion of melatonin was of similar magnitude at these two occasions. By contrast, the GH secretion was significantly lower the nights the healthy controls were given octreotide (GH AUC 22.6+/-5.4 mU/l x h during octreotide and 126.6+/-21.9 mU/l x h during saline; p<0.01). The patients with hypothyroidism also showed similar nocturnal melatonin secretion during octreotide and saline. Urinary excretion of melatonin also remained unchanged, as did GH secretion. The total nocturnal secretion of TSH was, however, significantly reduced by octreotide (TSH AUC 562+/-136 mU/l x h during octreotide and 851+/-185 mU/l x h during saline; p<0.05), thus suggesting that 100 microg of octreotide should be sufficient to inhibit also the pinealocytes if their function were regulated by somatostatin. Since exogenous somatostatin--in the form of octreotide--fails to influence nocturnal secretion and urinary excretion of melatonin in normal subjects and in patients with primary hypothyroidism, it is reasonable to assume that endogenous somatostatin may not be an important regulator of melatonin secretion in man. PMID- 10475151 TI - Homologies of the thyroid sodium-iodide symporter with bacterial and viral proteins. AB - We have demonstrated that Na+/I- symporter (NIS), a novel thyroid autoantigen, has local amino acid sequence homologies with the other thyroid autoantigens: Thyroglobulin (Tg), thyroid peroxidase (TPO) and thyrotropin receptor (TSH-R). These homologies concern the 4th, 5th, 6th extracellular loop and the beginning of the intracellular tail. We have expanded our studies and found that there are significant local homologies with other 11 proteins, most of them of bacterial or viral origin (e.g., Streptococcus or Herpes). These homologies concern the 2nd and 4th extracellular loop, and both the beginning and the end of the intracellular tail. These 11 homologies were retrieved by a computer-assisted search and extracted out of a database containing almost 300,000 amino acid sequences. These homologies were of magnitude greater than those concerning the three thyroid autoantigens [identities=51.1+/-7.3% vs 25.3+/-7.8% (mean+/-SD), p<0.001; similarities=70.6+/-10.7% vs 43.3+/-8.5%; p<0.001]. In addition, extensive, not local, homology was found with a number of unknown proteins from invertebrates (Drosophila melanogaster and Caenorhabditis elegans) and bacteria such as Bacillus subtilis and Xanthobacter. Previously, we had found that NIS has no extensive homology with Tg or TPO or TSH-R. This is the first demonstration of both extensive and local homologies between one thyroid autoantigen (NIS) and microbiological proteins. Taken together with data of the literature on the homologies between other thyroid antigens (Tg, TPO, TSH-R) and bacteria, the homologies we have now found reinforce the view that both bacterial and viral infections may trigger autoimmune thyroid diseases. PMID- 10475152 TI - Growth hormone bioactivity and immunoactivity in tall children. AB - In subjects with constitutional tall stature, both low and high GH response to stimulation tests have been observed when measured by commercial kits. To investigate the reason for these conflicting results, we evaluated growth hormone (GH) secretion using different assays as well as GH-binding protein and insulin like growth factor-I (IGF-I) concentrations in tall children. Serum samples were collected from 22 prepubertal constitutionally tall children, aged 2.87-13.25 years, during two pharmacological tests to evaluate serum GH levels measured by both immunofluorometric assay (IFMA) and the Nb2 cell bioassay. Serum IGF-I values were evaluated by RIA. Circulating low affinity (LA) and high affinity (HA) GH-binding proteins (GHBPs) were evaluated by FPLC gel filtration. Considering the highest serum GH levels as measured by IFMA, the 22 tall subjects were divided into two groups: group A including 16 children with blunted serum GH peak levels (5.78+/-0.68 ng/ml) and group B including 6 subjects with normal serum GH peak values (15.73+/-1.56 ng/ml). No differences were observed in serum GH peak levels as measured by the Nb2 cell bioassay between group A (14.77+/-1.54 ng/ml) and group B (16.03+/-1.96 ng/ml), and between both groups and 11 age-and sex-matched controls (12.25+/-1.19 ng/ml). In group A, the Nb2 cell bioassay/IFMA ratio of serum GH peak levels (0.29+/-0.08) was significantly higher (p<0.05) than in group B (0.07+/-0.01). No differences were found in serum LA-GHBP and HA GHBP as well as in IGF-I concentrations between the 16 patients of group A and the 6 of group B. Likewise, no difference in auxological parameters was found between the two groups. The biological activity of GH evaluated using the Nb2 cell bioassay is similar in tall children with a low GH response as measured by IFMA in comparison with those with a normal GH response, and is in agreement with both the auxological data and serum IGF-I concentrations. PMID- 10475153 TI - Co-existence of osteogenesis imperfecta and hyperparathyroidism. AB - Osteogenesis imperfecta (OI) and hyperparathyroidism (HTP) are disorders affecting the skeletal system and calcium metabolism not evidently related to one another. We report a case in which both OI and HPT were present. Our female patient presented with hypercalcaemia (S-Ca2+ 1.59 mmol/l; normal range 1.15 1.30) and 4-gland parathyroid hyperplasia at 30 years of age. Since her first year she had fractures, blue sclera, hypermobile joints, short stature (height 1.51 m, weight 49.5 kg) but normal hearing, and dentiogenesis imperfecta (tooth disease caused by defective formation of dentin) was absent. This patient bears many similarities with the 5 patients reported previously but it is the only patient, to our knowledge, with OI and early onset of HPT (30 year old female). We have found the OI to be type 1. A minor improvement of the rate of bone turnover 10 months after parathyroidectomy indicates the HPT to be primary and suggests the OI type 1 and pHPT to be two different calcium metabolic diseases incidentally occurring in the same patient. PMID- 10475154 TI - Pheochromocytoma and sub-clinical Cushing's syndrome during pregnancy: diagnosis, medical pre-treatment and cure by laparoscopic unilateral adrenalectomy. AB - The coexistence of pheochromocytoma and primary adrenal Cushing's syndrome of the same adrenal gland has rarely been reported. We describe here the case of a female patient presenting with mild Cushing's stigmata, hypertension and diabetes mellitus in whom we diagnosed a pheochromocytoma of the left adrenal gland with coexisting non-ACTH-dependent cortisol hypersecretion. While hormonal work-up was still in progress, the patient became pregnant and wanted to carry her pregnancy to full-term. A laparoscopic adrenalectomy in the 17th week of gestation was decided upon and the patient accordingly prepared for surgery by pre-treatment with phenoxybenzamine. Successful surgery--the first ever reported laparoscopic resection of a pheochromocytoma in pregnancy--without perioperative complications was performed under general anesthesia, with the patient receiving peri- and post operative hydrocortisone substitution. Pathohistological examination revealed a pheochromocytoma with positive immunostaining for interleukin-6 (IL-6) and negative immunostaining for ACTH, vasoactive intestinal polypeptide (VIP) and cytochrome P450, and with no signs of malignancy. A paracrine stimulation of the ipsilateral adrenal cortex by IL-6 produced by the pheochromocytoma, leading to cortical hyperplasia and subclinical Cushing's syndrome, is suggested by the positive immunostaining for IL-6 and the MRI findings. Post-operatively, secondary adrenal insufficiency ensued, necessitating continuing hydrocortisone replacement over 12 months. Hypertension resolved after surgery, and diabetes after the uncomplicated vaginal delivery at term. PMID- 10475155 TI - Diabetes insipidus and increased serum levels of leptin and lactate-dehydrogenase (LDH) in an adolescent boy with a primary intracranial germinoma. Case report and an endocrinological revaluation of literature. AB - A 16-year-old boy presented with a four-month history of polyuria-polydipsia and a diplopia which had reverted after treatment. The neuroimaging studies performed had been strongly suggestive of an optic nerve glioma, while endocrinological investigation (beta-hCG 420 IU/L) has lead to the correct diagnosis later confirmed at the immunohystochemical analysis performed at biopsy. The high serum level of hCG was unaffected by bromocriptine nor octreotide, while the PRL level (80.0 microg/L) was reduced only by bromocriptine. Among the several tumor markers which may be secreted by such lesions, ours is the first reported case of an elevation of serum LDH for a primary intracranial germinoma. Moreover, the elevated value of serum leptin reported by us might be due to the insensitivity of the hypothalamic structures to endogenous leptin. PMID- 10475156 TI - Beta-casein in cow's milk: a major antigenic determinant for type 1 diabetes? PMID- 10475158 TI - "The dwarf" (Sebastiano de Morra). Velazquez. Sevilia (1599)-Madrid (1660), Spain. PMID- 10475157 TI - Controversial topics series: milk proteins and diabetes. PMID- 10475159 TI - Separate mechanisms for short- and long-term memory. AB - It has been assumed for over a century that short-term memory (STM) processes are in charge of cognition while long-term memory (LTM) is being formed, a process that takes hours. A major question is whether STM is merely a step towards LTM, or a separate entity. Recent experiments have shown that many treatments with specific molecular actions given into the hippocampus, entorhinal or parietal cortex immediately after one-trial avoidance training can effectively block STM without affecting LTM formation. This shows that STM and LTM involve separate mechanisms. Some treatments even affect STM and LTM in opposite directions. Others, however, influence both memory types similarly, suggesting links between the two both at the receptor and at the post-receptor level. Drug effects on working memory (WM) were also studied. In some brain regions WM is affected by receptor blockers that alter either STM or LTM; in others it is not. This suggests links between the three memory types at the receptor level. The anterolateral prefrontal cortex is crucial for WM and LTM but is not involved in STM. The hippocampus, entorhinal and parietal cortex are crucial for the three types of memory, in some cases using different receptors for each. The amygdala is not involved in WM or STM, but it plays a key role in the modulation of the early phase of LTM. PMID- 10475160 TI - 6-hydroxydopamine lesion in the A8 cell group of cats produces a short-lasting decreased accuracy in goal-directed forepaw-movements. AB - Recently, feline studies have shown that a lesion in the retrorubral area, which includes the dopaminergic A8 cell group, produces motor programming deficits inherent to a hypofunction of the A9 system. A hypofunction in the striatal terminal area of A9 fibers, in turn, is known to produce a hypofunction of its first-order output station, namely the substantia nigra pars reticulata (SNR). The integrity of the SNR allows animals to execute (1) 'postural adjustments that rely on proprioceptive stimuli that originate in body parts at rest' and (2) 'non externally guided' targeting movements. In view of these considerations, the (dys)function of the SNR of cats with a bilaterally 6-hydroxydopamine lesion of A8 cells in the retrorubral area was tested in an experimental set-up that allows the assessment of changes in these functions. The A8 lesion produced: (a) a short lasting increase in the number of accurate targeting movements as well as an increase in the time required for the collection of six pellets: these deficits disappeared 4-7 days after the lesion; (b) a long-lasting disappearance of (1) 'postural adjustments that rely on proprioceptive stimuli that originate in body parts at rest' and (2) 'non-externally guided targeting movements'; and (c) a long-lasting display of a new strategy that allowed the lesioned cat to collect its pellets despite of its other deficits. These data led to the conclusion that a lesion of A8 cells even disrupts the function of the SNR, being one of the outputstations of the A8 cell group. PMID- 10475161 TI - GABA(A), D1, and D5, but not progestin receptor, antagonist and anti-sense oligonucleotide infusions to the ventral tegmental area of cycling rats and hamsters attenuate lordosis. AB - In hamsters, progesterone (P) in the hypothalamus and ventral tegmental area (VTA) is necessary for receptivity; in rats, hypothalamic P induces receptivity and midbrain P further enhances it. How P exerts its effects in the VTA on lordosis is of interest because few estrogen-induced P receptors (PRs) have been identified there. Sexual receptivity of rats and hamsters is enhanced when P's actions in the VTA are restricted to the membrane and when the gamma-aminobutyric acid (GABA)A agonist, muscimol, is infused into the VTA, but attenuated with infusions of the GABA(A) antagonist, bicuculline. The dopamine (DA) agonist. SKF38393, rapidly enhances receptivity when infused intravenously; this effect can be blocked by both DA receptor (DR) and PR antagonists. This study investigated the importance of PRs, glutamic acid decarboxylase (GAD), the enzyme responsible for GABA production, GABA(A) receptors (GBRs), and DRs in the VTA of cycling rats and hamsters for the expression of lordosis. Proestrous and diestrous animals implanted with bilateral VTA cannulae were pre-tested for receptivity, infused with either an antagonist (RU38486 (20 microg), bicuculline (100 ng), SCH23390 (100 ng)), anti-sense oligonucleotide (against PR (250 ng), GAD (500 ng), D1 (500 ng), D5 (250 ng)), or control infusions to each cannulae and re-tested. Vehicle and scrambled oligonucleotides were infused as controls and elicited similar effects. Antagonists of GBRs and DRs significantly reduced lordosis on post-tests compared to the PR antagonist and control conditions in rats and hamsters. Lordosis was significantly reduced, compared to controls, only by anti-sense oligonucleotides for GAD and D1- and D5-DR subtypes. These data suggest that in the VTA GABAergic and dopaminergic neurons may be more important in the mediation of sexual receptivity than neurons containing intracellular PRs. PMID- 10475162 TI - Dopamine antagonism in a novel-object recognition and a novel-object place conditioning preparation with rats. AB - Access to novel objects, similar to drugs of abuse, can enhance a place preference in rats. In the present experiments, the dopamine D1 receptor antagonist SCH-23390 blocked an increase in place preference conditioned by access to novel objects at doses that did not interfere with object interaction (0.01 and 0.03 mg/kg) or produce a place aversion in controls. However, eticlopride, a D2/D3 dopamine receptor antagonist, only blocked the conditioned increase in place preference at a dose (0.3 mg/kg) that impaired object interaction. In contrast, neither SCH-23390 nor eticlopride blocked preference for the novel object in an object recognition task at doses that did not interfere with object interaction. These experiments provide further evidence that the neural processes controlling learned associations between novel stimuli and the environment overlap with drugs of abuse. PMID- 10475163 TI - Complete locomotor recovery following corticospinal tract lesions: measurement of ground reaction forces during overground locomotion in rats. AB - The corticospinal tract is a prominent descending pathway in rodents which is thought to be involved in motor control. The purpose of this study was to investigate whether the lesions of the corticospinal tract affected overground locomotion in rats using a method of assessment which is relatively novel in this field, the evaluation of ground reaction forces. Ground reaction forces, i.e. the forces acting through the limbs on the ground, are a non-invasive, sensitive and quantitative method with which to assess gait compensation and the function of individual limbs during locomotion. We compared the three-dimensional ground reaction forces produced by locomoting rats with a unilateral corticospinal tract transection to those of control rats. Corticospinal-lesioned animals showed transient locomotor deficits 24-48 h after the lesion which quickly recovered to normal symmetrical locomotion. The initial locomotor deficits consisted of an asymmetric gait during which abnormal braking forces were produced during the dual contact time of the forelimb contralateral to the lesion (the impaired forelimb) and the ipsilateral (unimpaired) hindlimb. Normal forces were produced during the dual contact time of the ipsilateral (unimpaired) forelimb and the contralateral (impaired) hindlimb. The presence of the initial deficits may be a result of inflammation in the area of the lesion or may reflect a loss of normal locomotor contribution of the corticospinal tract. If the latter, the nature of the locomotor deficit suggests that (a) the forelimb may be more influenced by corticospinal lesions than is the hindlimb and/or (b) that the asymmetric gait produced is a general compensatory response to unilateral CNS injury in a quadruped. Complete recovery from corticospinal tract transection provides unequivocal evidence that input from the corticospinal tract is not essential for normal overground locomotion in the rat. PMID- 10475164 TI - Inhibitory effects of ginseng total saponin on nicotine-induced hyperactivity, reverse tolerance and dopamine receptor supersensitivity. AB - A single administration of a low dose of nicotine produced hyperactivity in mice. A repeated administration of nicotine developed reverse tolerance to the ambulation-accelerating activity of nicotine and also developed postsynaptic dopamine (DA) receptor supersensitivity. The development of reverse tolerance was evidenced by an increased ambulatory response to nicotine, and the development of postsynaptic DA receptor supersensitivity was evidenced by the enhanced response in ambulatory activity to apomorphine, a DA receptor agonist. Administration of ginseng total saponin (GTS) prior to and during the nicotine treatment in mice inhibited not only nicotine-induced hyperactivity and reverse tolerance, but also postsynaptic DA receptor supersensitivity in nicotine-induced reverse tolerant mice. These results suggest that inhibition by GTS of nicotine-induced hyperactivity and reverse tolerance may be closely related with the inhibition of the dopaminergic activation induced by nicotine and that the development of nicotine-induced reverse tolerance may be associated with enhanced DA receptor sensitivity. PMID- 10475165 TI - Effects of environmental enrichment on cognitive function and hippocampal NGF in the non-handled rats. AB - In this study we examine whether exposure to differential housing after weaning would counteract the effects of postnatal handling (H) or nonhandling (NH) treatment by affecting learning and memory processes in young rats. In addition, we seek to determine if experience in enriched environment would alter hippocampal nerve growth factor (NGF) levels which is one of the factors known to be involved in the regulation of the survival and differentiation of developing basal forebrain neurones. Rats were either exposed to handling treatment, or left undisturbed starting day 1 after birth through day 21. After weaning on day 22, we exposed half of the H rats and half of the NH rats to environmental enrichment for 60 days. The other respective half of the rats was housed in isolated environmental condition (IC). Behavioural measures were taken in open field test, and spatial water maze test. Exposure to enriched environment following postnatal handling and nonhandling increased hippocampal NGF levels, and improved cognitive function in the both groups, with NH rats being more responsive to the effects of enrichment. Our results suggest that environmental enrichment has the potential to prevent or reduce the cognitive and neurochemical deficits in the adult animals associated with nonhandling. PMID- 10475166 TI - Long-term voluntary ethanol consumption affects neither spatial nor passive avoidance learning, nor hippocampal acetylcholine release in alcohol-preferring rats. AB - Long-term ethanol consumption in humans and laboratory animals is associated with morphological and functional alterations of brain structures involved in cognitive processes. In the present experiments, we assessed whether voluntary long-term consumption of ethanol by alcohol-preferring (sP) rats under free choice condition with water (also) caused alterations in memory performance and hippocampal acetylcholine (ACh) release in vivo. A group of sP rats were offered a 10% v/v ethanol solution in a free choice with water for 36 weeks; controls had only tap water available. After withdrawal of ethanol, rats were tested in one trial passive avoidance test and thereafter were trained in a food-reinforced radial arm maze task for 12 days. One day after the last session in the radial arm maze, rats were implanted with a microdialysis probe in the dorsal hippocampus and dialysate concentrations of ACh were measured. No significant differences were observed between sP drinking and control rats in retention latencies in the passive avoidance test, in radial arm-maze performance or in basal levels of hippocampal ACh release. These results show that long-term ethanol consumption by sP rats is not associated with cognitive impairments or with alterations in the hippocampal cholinergic function. To the extent that chronic ethanol intoxication can be considered a causal factor in the development of memory and neurochemical alterations, these results suggest that sP rats self regulate ethanol consumption so as to avoid intoxication. These findings may challenge the notion that sP rat lines can be considered a valid model of human alcoholism. PMID- 10475167 TI - Effects of hippocampal lesions on spatial operant discrimination in pigeons. AB - In experiment 1, pigeons were trained on spatial or color autodiscrimination. Presentation of one of two keys or one of two colors was followed by food presentation. However, the other side of the keys or the other color was not. The hippocampal lesions disturbed the acquisition of spatial discrimination but not of color discrimination. In experiment 2, pigeons were preoperatively trained the spatial autodiscrimination, then received the hippocampal lesions. The subjects maintained the discrimination. These results suggest that the avian hippocampus plays a crucial role in acquisition of spatial discrimination. PMID- 10475169 TI - Time structure of a goal-directed bimanual skill and its dependence on task constraints. AB - The aim of the study was to elucidate the underlying principles of bimanual coordination and to establish quantitative coordination criteria. Healthy human subjects were instructed to open a loaded drawer with the left hand and to grasp, lift and reinsert with the right hand a small peg in the drawer recess. This bimanual goal-oriented task was executed promptly and consistently after a few trials. The temporal structure of the individual limb actions was assessed for computing interlimb synchronization and temporal correlation. In all subjects, both hands were well synchronized at the goal with high intermanual correlation in reaching the goal (event times of drawer opening and grasping the peg). This temporal goal-invariance was independent of movement speed and of the highly variable timing of the individual hands and persisted when subjects were blindfolded. Unilateral loading of the pulling hand and cutaneous anesthesia of the left index finger and thumb used for grasping the drawer handle significantly increased the pull-phase. This slowing of the left hand was matched by an adaptive delay of the right non-disturbed hand, thus preserving goal invariance. As a working hypothesis, we propose that multimodal sensory signals generated in the leading arm be transmitted centrally to re-parameterize the non-disturbed arm. PMID- 10475168 TI - Stress- and corticosteroid-induced modulation of the locomotor response to morphine in rats. AB - Stress alters the sensitivity to drugs of abuse and is, therefore, considered to be an important contributory factor to drug-seeking behaviour. There is only a limited amount of information available on stress-induced alterations in the behavioural response to opioids. We thus evaluated the influences of different stressors (restraint, handling, social defeat) on the locomotor effects induced by morphine. Further the importance of additional factors such as the number of stress events or the delay between stress and locomotor testing on stress-induced alterations were evaluated. Since these modulatory effects of stress on the locomotor effects of morphine might be mediated via the release of endogenous corticosteroids we also tested the influence of repeated intermittent and chronic administration of corticosterone (CORT) and the synthetic corticosteroid dexamethasone (DEX) on the locomotor response to morphine. Enhanced morphine induced locomotor activity was observed in response to the repeated application (three times) of all stressors: restraint, handling and social defeat. An augmentation of the locomotor effects of a low (1 and 5 mg/kg) but not of a high dose (10 mg/kg) of morphine was seen after three, but not after one stress event. In addition, the repeated application of restraint stress (three times) resulted in an augmentation of morphine-induced locomotor stimulation 3 days, but not 1 or 10 days , after the last stress event. Similarly the repeated intermittent and chronic administration of corticosteroids, in particular of DEX, increased morphine's efficacy in stimulating locomotor activity. Our results show that stress is able to alter the locomotor stimulant effects of morphine in rats--a phenomenon called stress-induced behavioural sensitization. Moreover, these stress-induced alterations depend upon temporal factors such as number of stress events and the interval between stress and locomotor testing. Further, stress induced CORT-release seems to be involved in stress-induced behavioural sensitization to morphine. PMID- 10475171 TI - Intrahippocampal injections of cysteamine improve the retention of a bar-pressing task in mice. AB - Cysteamine was used as a tool aimed at investigating the role of central somatostatin (SS-14) and was shown to modulate learning in a task-dependent manner. However, direct arguments have not yet been provided to support the hypothesis that impairments and facilitation of learning produced by cysteamine are both mediated by the hippocampus. Mice were given daily intrahippocampal injections of artificial cerebrospinal fluid (CSF) or cysteamine at doses of either 2.5 microg/0.2 microl or 25 microg/0.2 microl 1 h prior to each learning session of a bar-pressing task, for which the acquisition was previously shown to be improved by systemic injections. The results showed that, with respect to CSF, the mice injected with cysteamine learned the bar pressing task faster whereas no evidence of changes in locomotor activity was provided. Moreover, the results showed that retention was specifically increased in the two groups injected with cysteamine. It is argued that the action of cysteamine on the hippocampus is sufficient to modulate specifically learning-memory processes in a task-dependent manner. In conclusion, the blockade of some hippocampal information processing function by cysteamine is discussed to understand the bidirectional effects of drugs on learning and memory. PMID- 10475170 TI - Involvement of the serotonergic neuronal system in phencyclidine-induced place aversion in rats. AB - The possible involvement of the serotonergic neuronal system in aversive motivation produced by phencyclidine [1-(1-phenylcyclohexyl)piperidine; PCP] was investigated using a place-conditioning paradigm in rats. PCP (4 mg/kg, i.p.) produced place aversion in this task as reported previously (Kitaichi K, Noda Y, Hasegawa T, Furukawa H, Nabeshima T. Acute phencyclidine induces aversion, but repeated phencyclidine induces preference in the place conditioning test in rats. Eur J Pharmacol 1996;318:7-9). The blockade of serotonin2A (5-HT2A) receptors using the antagonist ritanserin (3 and 10 mg/kg, p.o.) significantly attenuated this aversive property of PCP whereas lesions of serotonergic neurons using 5,7 dihydroxytryptamine (5,7-DHT, 100 microg/animal, i.c.v.) failed to affect it. Repeated PCP treatment (10 mg/kg, i.p. for 14 days), which is enough to diminish the stereotyped 5-HT2A receptor-mediated head-twitch behavior, also decreased the place aversion. These results suggest that the serotonergic neuronal system, specifically the 5-HT2A receptor, may play a critical role in producing PCP induced place aversion. PMID- 10475172 TI - Two different properties of short- and long-term memory. AB - Rats were trained in one-trial step-down inhibitory avoidance and tested 1.5-9 h later or 1-4 days later in order to evaluate short- and long-term memory respectively. No extinction was observed when the animals were tested repeatedly at 1.5, 3, 4.5 and 6 h; when they were tested repeatedly at 9, 24, 48, 72 and 96 h, there was partial extinction. Exposure to an open field for 2 min, 1 h after training did not affect retention scores of animals tested at 1.5 or 3 h, had a slight amnestic effect on groups tested at 4.5 or 6 h, and markedly reduced retention scores in animals tested at 9 or 24 h from training. Thus, there were two clear behavioural differences between memory measured in the first few hours or in the following few days, which suggests that both involve different mechanisms. PMID- 10475173 TI - The spectrum of disability. PMID- 10475174 TI - Contemporary approach to management of unstable angina. PMID- 10475175 TI - Future of reperfusion therapy for acute myocardial infarction. PMID- 10475176 TI - Transmission of drug-resistant strains of HIV-1: unfortunate, but inevitable. PMID- 10475177 TI - Typhoid fever and childhood vaccine strategies. PMID- 10475178 TI - Efficacy of sunscreens in protection against skin cancer. PMID- 10475179 TI - An ideal husband in the wild(e)? PMID- 10475180 TI - Long-term low-molecular-mass heparin in unstable coronary-artery disease: FRISC II prospective randomised multicentre study. FRagmin and Fast Revascularisation during InStability in Coronary artery disease. Investigators. AB - BACKGROUND: Short-term treatment with subcutaneous low-molecular-mass heparin in addition to aspirin is effective in unstable coronary-artery disease. We assessed the efficacy of long-term treatment with dalteparin in patients managed with a non-invasive treatment strategy. METHODS: 2267 patients from three Scandinavian countries (median age 67 years, 68% men) with unstable coronary-artery disease were randomly assigned to continue double-blind subcutaneous dalteparin twice daily or placebo for 3 months, after at least 5 days' treatment with open-label dalteparin. The composite primary endpoint was death or myocardial infarction. Analysis was by intention to treat. FINDINGS: During the 3 months of double-blind treatment, there was a non-significant decrease in the composite endpoint of death or myocardial infarction of 6.7% and 8.0% in the dalteparin and placebo groups, respectively (risk ratio 0.81 [95% CI 0.60-1.10], p=0.17). At 30 days, this decrease was significant (3.1 vs 5.9%, 0.53 [0.35-0.80]; p=0.002). In the total cohort there was at 3 months a decrease in death, myocardial infarction, or revascularisation (29.1 vs 33.4%, 0.87 [0.77-0.99]; p=0.031). The initial benefits were not sustained at 6-month follow-up. INTERPRETATION: Long-term dalteparin lowers the risk of death, myocardial infarction, and revascularisation in unstable coronary-artery disease at least during the first month of therapy. These early protective effects could be used to lower the risk of events in patients waiting for invasive procedures. PMID- 10475181 TI - Invasive compared with non-invasive treatment in unstable coronary-artery disease: FRISC II prospective randomised multicentre study. FRagmin and Fast Revascularisation during InStability in Coronary artery disease Investigators. AB - BACKGROUND: In unstable coronary-artery disease early invasive procedures are common, despite lack of evidence for the superiority of this approach. We compared an early invasive with a non-invasive treatment strategy in unstable coronary-artery disease. METHODS: In a prospective randomised multicentre study, we randomly assigned 2457 patients in 58 Scandinavian hospitals (median age 66 years, 70% men) an early invasive or non-invasive treatment strategy with placebo controlled long-term low-molecular-mass heparin (dalteparin) for 3 months. Coronary angiography was done within the first 7 days in 96% and 10%, and revascularisation within the first 10 days in 71% and 9% of patients in the invasive and non-invasive groups, respectively. We followed up patients for 6 months. Analysis was by intention to treat. FINDINGS: After 6 months there was a decrease in the composite endpoint of death or myocardial infarction of 9.4% in the invasive group, compared with 12.1% in the non-invasive group (risk ratio 0.78 [95% CI 0.62-0.98], p=0.031). There was a significant decrease in myocardial infarction alone (7.8 vs 10.1%, 0.77 [0.60-0.99]; p=0.045) and non-significantly lower mortality (1.9 vs 2.9%, 0.65 [0.39-1.09]; p=0.10). Symptoms of angina and re-admission were halved by the invasive strategy. Results were independent of the randomised dalteparin treatment. The greatest advantages were seen in high risk patients. INTERPRETATION: The early invasive approach should be the preferred strategy in most patients with unstable coronary-artery disease who have signs of ischaemia on electrocardiography or raised biochemical markers of myocardial damage. PMID- 10475182 TI - Single-bolus tenecteplase compared with front-loaded alteplase in acute myocardial infarction: the ASSENT-2 double-blind randomised trial. AB - BACKGROUND: Bolus fibrinolytic therapy facilitates early efficient institution of reperfusion therapy. Tenecteplase is a genetically engineered variant of alteplase with slower plasma clearance, better fibrin specificity, and high resistance to plasminogen-activator inhibitor-1. We did a double-blind, randomised, controlled trial to assess the efficacy and safety of tenecteplase compared with alteplase. METHODS: In 1021 hospitals, we randomly assigned 16,949 patients with acute myocardial infarction of less than 6 h duration rapid infusion of alteplase (< or = 100 mg) or single-bolus injection of tenecteplase (30-50 mg according to bodyweight). All patients received aspirin and heparin (target activated partial thromboplastin time 50-75 s). The primary outcome was equivalence in all-cause mortality at 30 days. FINDINGS: Covariate-adjusted 30 day mortality rates were almost identical for the two groups--6.18% for tenecteplase and 6.15% for alteplase. The 95% one-sided upper boundaries of the absolute and relative differences in 30-day mortality were 0.61% and 10.00%, respectively, which met the prespecified criteria of equivalence (1% absolute or 14% relative difference in 30-day mortality, whichever difference proved smaller). Rates of intracranial haemorrhage were similar (0.93% for tenecteplase and 0.94% for alteplase), but fewer non-cerebral bleeding complications (26.43 vs 28.95%, p=0.0003) and less need for blood transfusion (4.25 vs 5.49%, p=0.0002) were seen with tenecteplase. The rate of death or non-fatal stroke at 30 days was 7.11% with tenecteplase and 7.04% with alteplase (relative risk 1.01 [95% CI 0.91 1.13]). INTERPRETATION: Tenecteplase and alteplase were equivalent for 30-day mortality. The ease of administration of tenecteplase may facilitate more rapid treatment in and out of hospital. PMID- 10475183 TI - Daily sunscreen application and betacarotene supplementation in prevention of basal-cell and squamous-cell carcinomas of the skin: a randomised controlled trial. AB - BACKGROUND: The use of sunscreens on the skin can prevent sunburn but whether long-term use can prevent skin cancer is not known. Also, there is evidence that oral betacarotene supplementation lowers skin-cancer rates in animals, but there is limited evidence of its effect in human beings. METHODS: In a community-based randomised trial with a 2 by 2 factorial design, individuals were assigned to four treatment groups: daily application of a sun protection factor 15-plus sunscreen to the head, neck, arms, and hands, and betacarotene supplementation (30 mg per day); sunscreen plus placebo tablets; betacarotene only; or placebo only. Participants were 1621 residents of Nambour in southeast Queensland, Australia. The endpoints after 4.5 years of follow-up were the incidence of basal cell and squamous-cell carcinomas both in terms of people treated for newly diagnosed disease and in terms of the numbers of tumours that occurred. Analysis of the effect of sunscreen was based only on skin cancers that developed on sites of daily application. All analyses were by intention to treat. FINDINGS: 1383 participants underwent full skin examination by a dermatologist in the follow-up period. 250 of them developed 758 new skin cancers during the follow-up period. There were no significant differences in the incidence of first new skin cancers between groups randomly assigned daily sunscreen and no daily sunscreen (basal cell carcinoma 2588 vs 2509 per 100,000; rate ratio 1.03 [95% CI 0.73-1.46]; squamous-cell carcinoma 876 vs 996 per 100,000; rate ratio 0.88 [0.50-1.56]). Similarly, there was no significant difference between the betacarotene and placebo groups in incidence of either cancer (basal-cell carcinoma 3954 vs 3806 per 100,000; 1.04 [0.73-1.27]; squamous-cell carcinoma 1508 vs 1146 per 100,000; 1.35 [0.84-2.19]). In terms of the number of tumours, there was no effect on incidence of basal-cell carcinoma by sunscreen use or by betacarotene but the incidence of squamous-cell carcinoma was significantly lower in the sunscreen group than in the no daily sunscreen group (1115 vs 1832 per 100,000; 0.61 [0.46 0.81]). INTERPRETATION: There was no harmful effect of daily use of sunscreen in this medium-term study. Cutaneous squamous-cell carcinoma, but not basal-cell carcinoma seems to be amenable to prevention through the routine use of sunscreen by adults for 4.5 years. There was no beneficial or harmful effect on the rates of either type of skin cancer, as a result of betacarotene supplementation. PMID- 10475184 TI - Transmission of antiretroviral-drug-resistant HIV-1 variants. AB - BACKGROUND: Resistance of HIV-1 to antiretroviral drugs is the main cause of antiretroviral-treatment failure. We assessed the transmission of drug-resistant variants among individuals with primary HIV-1 infection. METHODS: Population based sequencing of the viral reverse-transcriptase and protease genes derived from plasma viral RNA was done in 82 consecutive individuals with documented primary HIV-1 infection from January, 1996, to July, 1998. Phenotypic resistance to protease inhibitors was assessed by recombinant virus assay in individuals with two or more mutations associated with resistance to protease inhibitors. FINDINGS: Zidovudine-resistance mutations were detected in seven (9%) of 82 individuals. Mutations associated with resistance to other reverse-transcriptase inhibitors (RTIs) were detected in two individuals. Primary-resistance mutations associated with protease inhibitors (V82A, L90M) were detected in three (4%) of 70 individuals; two of these had also RTI-resistance mutations. Decreased sensitivity to three or four protease inhibitors was seen in three individuals, one of whom was infected with HIV-1 variants that harboured 12 mutations associated with resistance to multiple RTI and protease inhibitors. INTERPRETATION: To introduce the best antiretroviral treatment, resistance testing should be done in recently HIV-1-infected individuals. PMID- 10475186 TI - Collapse with a streptococcal infection. PMID- 10475185 TI - Typhoid fever in children aged less than 5 years. AB - BACKGROUND: Calculation of the incidence of typhoid fever during preschool years is important to define the optimum age of immunisation and the choice of vaccines for public-health programmes in developing countries. Hospital-based studies have suggested that children younger than 5 years do not need vaccination against typhoid fever, but this view needs to be re-examined in community-based longitudinal studies. We undertook a prospective follow-up study of residents of a low-income urban area of Delhi, India, with active surveillance for case detection. METHODS: A baseline census was undertaken in 1995. Between Nov 1, 1995, and Oct 31, 1996, we visited 8172 residents of 1820 households in Kalkaji, Delhi, twice weekly to detect febrile cases. Blood samples were obtained from febrile patients, and those who tested positive for Salmonella typhi were treated with ciprofloxacin. FINDINGS: 63 culture-positive typhoid fever cases were detected. Of these, 28 (44%) were in children aged under 5 years. The incidence rate of typhoid per 1000 person-years was 27.3 at age under 5 years, 11.7 at 5-19 years, and 1.1 between 19 and 40 years. The difference in the incidence of typhoid fever between those under 5 years and those aged 5-19 years (15.6 per 1000 person-years [95% CI 4.7-26.5]), and those aged 19-40 years (26.2 [16.0 36.3]) was significant (p<0.001 for both). The difference between the incidence of typhoid at 5-19 years and the incidence at 19-40 years was also significant (10.6 [6.3-14.8], p<0.001). Morbidity in those under 5 and in older people was similar in terms of duration of fever, signs and symptoms, and need for hospital admission. INTERPRETATION: Our findings challenge the common view that typhoid fever is a disorder of school-age children and of adults. Typhoid is a common and significant cause of morbidity between 1 and 5 years of age. The optimum age of typhoid immunisation and the choice of vaccines needs to be reassessed. PMID- 10475187 TI - Acute effect of nitric oxide on Raynaud's phenomenon in scleroderma. AB - Intra-arterial infusions of L-arginine and sodium nitroprusside significantly decreased the occurrence of laboratory-induced Raynaud's phenomenon in scleroderma patients. Raising the concentration of nitric oxide may be of therapeutic value in this population. PMID- 10475188 TI - Acne fulminans in late-onset congenital adrenal hyperplasia. AB - Acne may be the only clinical sign of androgen excess in men. We report a boy with acne fulminans and androgen excess due to late-onset congenital adrenal hyperplasia. PMID- 10475189 TI - Sublingual therapy for cobalamin deficiency as an alternative to oral and parenteral cobalamin supplementation. AB - Effectiveness of sublingual cobalamin-replacement therapy was studied in 18 people with cobalamin deficiency. Administration was efficacious and convenient, and compliance was high. PMID- 10475190 TI - Decrease of vancomycin-resistant enterococci in poultry meat after avoparcin ban. AB - In Italy, 18 months after the ban of avoparcin, the percentage of poultry meat samples containing vanA gene-positive vancomycin-resistant enterococci fell from 14.6% to 8%. PMID- 10475191 TI - Trends in mortality from cancer in the European Union, 1955-94. AB - Over the past few years, age-standardised mortality rates have levelled off or declined for most cancer sites in the European Union, with the main exception of lung cancer in women. PMID- 10475192 TI - Cytotoxic T lymphocyte antigen-4 (CTLA-4) gene polymorphism confers susceptibility to thyroid associated orbitopathy. AB - Smoking and demographic variables are known to be risk factors for the development of thyroid-associated orbitopathy (TAO) among patients with Graves' hyperthyroidism, but a firm genetic basis for TAO has not been established. We show that the presence and severity of TAO are associated with an allele of the cytotoxic T lymphocyte antigen-4-gene. PMID- 10475193 TI - Osteopenia in patients with clinically silent coeliac disease warrants screening. AB - Bone-mineral density was measured in 19 clinically silent coeliac patients. The density was low, comparable with that in coeliac cases, and improved by gluten free diet. PMID- 10475194 TI - Ceasing of movement-disorder attacks immediately after the onset of pregnancy: possible effect of human chorionic gonadotropin. AB - In a woman with paroxysmal kinesiogenic choreoathetosis, attacks ceased within a few days after conception. An effect of human chorionic gonadotrophin is assumed, since this hormone decreased sodium currents and excessive action potential generation in an experimental approach. PMID- 10475196 TI - Braving the peaks? Be prepared. PMID- 10475195 TI - You are better off running than walking with acute vestibulopathy. AB - Patients with acute vestibular disorder balance better when running than when standing or walking slowly. We suggest that the automatic spinal locomotor programme suppresses destabilising vestibular input. PMID- 10475197 TI - Bakhsheesh culture contributes to Turkey's misery. PMID- 10475198 TI - UN condemns Taliban's continued human rights abuse in Afghanistan. PMID- 10475199 TI - UK inquiry opened into human trials of biological and chemical weapons. PMID- 10475200 TI - Inmates in Irish prisons face drug abuse and disease. PMID- 10475201 TI - Variant CJD fears prompt growing number of countries to ban British blood donations. PMID- 10475202 TI - Spain offers free HIV tests to pregnant women. PMID- 10475203 TI - Disability in cross-cultural perspective: rethinking disability. PMID- 10475204 TI - The myth of disability in developing nations. PMID- 10475205 TI - Disability and rehabilitation: a context for understanding the American Indian experience. PMID- 10475206 TI - Community-based disability services in the USA: a paediatric perspective. PMID- 10475207 TI - The role of the family: disability and rehabilitation in rural Jamaica. PMID- 10475208 TI - A clash of cultures: reflections of a physician with a disability. PMID- 10475209 TI - Nuclear medicine's role in infection and inflammation. AB - Nuclear medicine imaging techniques can help in patient evaluation where infectious and non-infectious inflammatory disorders are suspected. When selected and tailored to the clinical situation, most techniques already in use or available soon provide information with high sensitivity. However, almost all currently available techniques lack the specificity to discriminate between infectious and non-infectious inflammation. In undiagnosed fever, this non specificity may be an advantage since fever of unknown origin is caused by infection in only about 25% of cases, but in the postoperative patient the reliable differentiation between infection and sterile inflammation is highly relevant to clinical management. The range of radiopharmceuticals to investigate infectious and non-microbial inflammatory disorders is expanding and developments in protein/peptide chemistry and in labelling technology should lead to agents with very high specific activities. Nuclear medicine has to add specificity to its already high sensitivity if it is to distinguish both categories of inflammatory disorder. PMID- 10475210 TI - Bosnia and Herzegovina and Croatia: the medicalisation of the experience of war. PMID- 10475211 TI - Bosnia and Herzegovina and Croatia: the medicalisation of the experience of war. PMID- 10475212 TI - Bosnia and Herzegovina and Croatia: the medicalisation of the experience of war. PMID- 10475213 TI - Nimesulide and COX-2 inhibitors. PMID- 10475214 TI - A plea for preservation of the pelvic autonomic nerves. PMID- 10475215 TI - False-negative rates in sentinel-node in breast cancer. PMID- 10475216 TI - False-negative rates in sentinel-node in breast cancer. PMID- 10475217 TI - Diagnosis of relative adrenal insufficiency in critically ill patients. PMID- 10475218 TI - Long-term use of immunoglobulin infusion device. PMID- 10475219 TI - Maternal mortality and mode of delivery. PMID- 10475220 TI - Sclerosing pancreatocholangitis. PMID- 10475221 TI - Antibacterial activity of hyperforin from St John's wort. PMID- 10475222 TI - Antibacterial activity of hyperforin from St John's wort. PMID- 10475223 TI - Antibacterial activity of hyperforin from St John's wort. PMID- 10475224 TI - Late effects of transplantation. Gerard Socie, for the Late Effects Working Party of the European Group for Blood and Marrow Transplantation. PMID- 10475225 TI - Erythropoietic quality of maternal milk. PMID- 10475226 TI - Belgium and statistics on mortality. PMID- 10475227 TI - Use of CS spray in hypoglycaemia. PMID- 10475228 TI - Androstenedione and dehydroepiandrosterone for athletes. PMID- 10475229 TI - Is there a Hispanic race? PMID- 10475230 TI - A metered way of life. PMID- 10475231 TI - A metered way of life. PMID- 10475232 TI - Drugs and the art of Fred Tomaselli. PMID- 10475233 TI - US online pharmacies strive for respectability. PMID- 10475234 TI - The Nobel chronicles. 1969: Max Delbruck (1906-81); Salvador Luria (1912-91); and Alfred Hershey (1908-97). PMID- 10475235 TI - Use of monoclonal antibodies for the diagnosis and treatment of bladder cancer. AB - Although the management of cancer by exploiting properties distinguishing neoplastic and normal cells has always been an attractive concept, it was the development of hybridoma technology and the resulting tumor-associated monoclonal antibodies (MAbs) that offered new prospects for this strategy. Twenty years later, some of the applications of MAbs in oncology are now part of the everyday diagnosis and treatment, while others are the subject of intensive investigation. We reviewed the current applications of MAbs in the diagnosis and treatment of bladder cancer and attempted to put the issue into perspective, with particular presentation of their therapeutic potential. PMID- 10475236 TI - Immunohistochemical analysis of the D-type cyclin-dependent kinases Cdk4 and Cdk6, using a series of monoclonal antibodies. AB - Cellular signal transduction cascades triggered by mitogenic or antiproliferative cues eventually converge on a biochemical mechanism centered around the retinoblastoma tumor suppressor (pRb), the so-called RB pathway that governs G1 phase progression and guards the commitment to enter S phase. pRb, together with its immediate upstream regulators, the D-type cyclins, their partner cyclin dependent kinases Cdk4 and Cdk6, and the Cdk inhibitors, form a functional unit that is involved in major decisions about cellular fate, and whose components, including the proto-oncogenic cyclin D-dependent kinases, are commonly deregulated in many types of cancer. We report here the production and characterization of a series of 12 monoclonal antibodies (MAbs) that specifically recognize either Cdk4 or Cdk6. These antibodies are proving to be invaluable molecular probes for defining abundance, subcellular localization, binding partners, and ultimately the function(s) of these cell cycle-regulatory kinases. Localization of the target epitopes was mapped by peptide enzyme-linked immunoadsorbent assay (ELISA), and two antibodies recognizing sequences adjacent to N-terminus of Cdk4 can discriminate between the wild-type protein and the oncogenic, melanoma-associated R24C mutant of this kinase. Individual antibodies of our panel recognize distinct pools of Cdk4/6, a feature reflected by their differential applicability in immunoblotting, immunoprecipitation, kinase assays, and immunostaining including immunohistochemistry on archival paraffin-embedded tissue sections. Collectively, the antibodies described in this study provide the means for immunochemical analyses of the cyclin D-dependent kinases in human and animal cells, and represent useful molecular tools that should help better understand the biological roles of Cdk4 and Cdk6 in normal cell-cycle control, and their oncogenic activity in tumor cells. PMID- 10475237 TI - Inhibitory mechanism of anti-P30-52 monoclonal antibody against human immunodeficiency virus type 1 (HIV-1) multiplication in infected MT-4 cells. AB - We have studied the immunological role of human immunodeficiency virus type 1 (HIV-1) p17 because the p17 antibody titer is high in asymptomatic patients and decreases with disease progression. Previously we found that monoclonal antibody (MAb) reactive to the p17-derived peptide 30 to 52 amino acids in length, namely P30-52 MAb, had cross-reactivity to the third variable region of the envelope glycoprotein of HIV-1 (Env V3) and also inhibited the viral multiplication of the supernatant of HIV-1-infected MT-4 cells co-cultured with the MAb. The relation between the cross-reactivity of the P30-52 MAb and the inhibitory mechanism is not clear; however, P30-52 might be useful for the development of therapeutic and vaccination strategies. In the present study, we examined the suppressive mechanism of the P30-52 MAbs, and found that the copy number of HIV-1 RNA in HIV 1-infected MT-4 cells was not reduced by the addition of the P30-52 MAbs, and the expression of RNAs of p17 was slightly enhanced 3 hr after the infection, although that of Env V3 was the same as the control level. In contrast, the expression of cellular p17 DNA and p17 protein was reduced by the addition of the P30-52 MAbs. In conclusion, the P30-52 MAbs did not suppress the HIV-1 mRNA level in the infected cells, but might inhibit DNA synthesis, and consequently bring about a reduction of p17 protein synthesis and a decrease of infectivity of the supernatant. The results demonstrated that the P30-52 MAb could be used as immunotherapeutic substance for HIV-1. PMID- 10475238 TI - Expression and characterization of anti-NCA-95 scFv (CEA 79 scFv) in a prokaryotic expression vector modified to contain a Sfi I and Not I site. AB - The CEA 79 antibody has been used in bone marrow scintigraphy for the differential diagnosis of skeletal tumors and the evaluation of the bone marrow status of patients with various hematological disorders. The specific localization of radio-labeled CEA 79 antibody in bone marrow depends on its reactivity with NCA-95 (nonspecific cross-reacting antigen-95) present on the surface and in the cytosol of human granulocytes and myelopoietic cells. To make a CEA 79 scFv molecule that would be less immunogenic and more penetrating than the intact mouse immunoglobulin, we constructed a pRSET Sfi I/Not I expression vector. The scFv gene was then excised from a pCANTAB 5 E phage display vector by digestion with Sfi I and Not I and inserted into the pRSET Sfi I/Not I expression vector. Upon transformation of a BL21(DE3)pLysS strain of E. coli, CEA 79 scFv became expressed in inclusion bodies requiring a renaturation process for solubilization. The final yield of CEA 79 scFv was 5 mg per a liter of culture. The refolded CEA 79 scFv exhibited an affinity (Kd = 2.1 x 10(-9) M) equivalent to that of the original CEA 79 antibody (K(d) = 3.3 x 10(-9) M) and the same immunoreactivity to CEA and NCA-95 in Western blots and in immunohistochemical staining experiments. PMID- 10475239 TI - Characterization of epitopes on human interleukin-2 using phage displayed-peptide libraries: insights into antibody-peptide interactions. AB - We have characterized the binding epitopes of two monoclonal antibodies (MAbs) reacting with human Interleukin-2 (IL-2), using a phage display peptide library. The first antibody (CB-IL2.1) recognizes the sequence LSFL, amino acid 72 to amino acid 80, numbered in the IL-2. The second antibody (CB-IL2.2) binds the sequence TTFM (amino acids 101 to 104) located at the opposite site of the four helix bundle of IL-2. Enzyme-linked immunoadsorbent assay (ELISA) and Western blot using different IL-2 protein construct expressed in bacteria and phage display demonstrate the specificities of this antibody. The data presented here show that the antibodies characterized in this study are raised against linear epitopes and suggest that these epitope are accessible from the outside in the native IL-2 molecule. PMID- 10475240 TI - Selection of specific phage from display libraries: monoclonal antibody against VCS M13 helper phage coat protein III (gIIIp). AB - Screening of specific phage is often hampered by nonspecific binding either of the VCS M13 helper phage to the solid phase absorbent or to the polyclonal antibodies used for selection. The former is improved by increasing the stringency for selection. However, the available polyclonal anti-VCS M13 antibodies often react with immobilized antigen nonspecifically, making it difficult to distinguish between positive and negative clones. To improve this selection process, a monoclonal antibody (MAb) was produced which recognizes ligand-coat protein three (gIIIp) on the helper phage VCS M13. This MAb is highly sensitive and specific, and it is useful for selecting relevant clones. This reagent should find widespread application in identifying interactive clones from a variety of phage display libraries. PMID- 10475241 TI - Novel cross-reactive anti-idiotype antibodies with properties close to the human intravenous immunoglobulin (IVIg). AB - The most important link between the immune network theory and clinically useful therapies so far is the use of human intravenous immunoglobulin (IVIg) in the treatment of autoimmune diseases. Although still controversial, one of the main mechanisms that has been postulated for the in vivo effects of IVIg, is the selection of immune repertoires through idiotypic interactions. We describe here anti-idiotype IgG monoclonal antibodies (MAbs), which were obtained by immunization of syngeneic mice (Balb/c) with an anti-ganglioside antibody. These anti-idiotype MAbs show multiple idiotypic connections and share some of the properties of the IVIg pool. The antiidiotype (Ab2) MAbs B7 and 34B7 showed heterogeneous binding with the idiotypes of several anti-ganglioside antibodies, MAbs obtained from splenocytes of nonimmunized newborn mice, F(ab')2 fragments of IgG human myeloma proteins, and nonimmunoglobulin antigens. The recognition pattern of the B7 MAb to the idiotypes of human immunoglobulins was also studied using a phage display library obtained from the variable region genes of an asymptomatic AIDS patient and also F(ab')2 fragments obtained from an IVIg pool of healthy human donors. We also demonstrated that these MAbs produced some of the in vitro effects reported for the human IVIg pool, such as the inhibition of cell proliferation of human B and T cell lines and of normal human lymphocytes activated with different mitogens. Another striking property of the MAb B7 was its ability to induce a dose-dependent specific antibody T-cell response in vivo in syngeneic mice. Both anti-idiotype MAbs showed anti-metastatic effect in vivo when injected intravenously to mice inoculated with MB16-F10 melanoma cells. The antimetastatic effect of the antiidiotype MAbs was not observed in athymic mice inoculated with the same tumor. This kind of antibody can become an interesting tool for further exploration of the role of idiotypic network connections in the regulation of the immune system and to study the effects of interventions on network connectivity in experimental autoimmune disease, using a reagent better chemically defined than the IVIg pool. PMID- 10475242 TI - Use of a PPAR gamma-specific monoclonal antibody to demonstrate thiazolidinediones induce PPAR gamma receptor expression in vitro. AB - Troglitazone and rosiglitazone (BRL49653), members of the thiazolidinedione (TZD) class of antidiabetic drugs, are peroxisome proliferator-activated receptor gamma (PPARgamma) ligands that induce adipocyte differentiation and increase the expression of PPARgamma protein. Here, we report the characterization of a PPARgamma specific monoclonal antibody (MAb), PgammaA53.25, and its use to monitor PPARgamma expression in the noncommitted pluripotent murine mesenchymal stem cell line, C3H10T1/2, treated with TZDs. MAb PgammaA53.25 was raised against a region in the N-terminal domain of human PPARgamma shared by splice variants PPARgamma1 and PPARgamma2. It recognizes immunizing antigen in enzyme-linked immunoadsorbent assay (ELISA), and does not cross-react with the N-terminal domains of PPARalpha or PPARdelta. In Western blotting, PgammaA53.25 reacts with the immunizing antigen as well as distinct protein bands corresponding to the molecular weight of full length PPARgamma from C3H10T1/2 cells and rat tissue lysates. In fluorescent microscopy, PgammaA53.25 immunostains nuclei of C3H10T1/2 cells treated with PPARgamma ligands. The fluorescence intensity of the treated cells is TZD dose-dependent, and correlates with lipid accumulation consistent with adipogenesis. Based on these results, we propose that MAb PgammaA53.25 will be a useful tool for elucidating the role of PPARgamma in fatty acid metabolism and adipocyte differentiation. PMID- 10475243 TI - Production and characterization of monoclonal antibodies to the steroid receptor coactivator AIB1. AB - The steroid coactivator AIB1 (Ampified In Breast cancer 1) is a member of the SRC 1 family of transactivation coactivators that is amplified in about 7% of breast tumor samples. Hybridomas were established that secreted monoclonal antibodies (MAbs) to a recombinant glutathione-S-transferase (GST) fusion protein expressing the steroid receptor interacting domain of AIB1 (GST-AIB.T1:a.a. 605-1294). Four MAbs from these hybridomas were characterized and designated AX15.1, AX15.2, AX15.3, and AX15.4. The MAbs were shown to be specific to AIB1 and did not cross react with two similar coactivators SRC-1 and TIF2 as shown in Western blot analysis and enzyme-linked immunoadsorbent assay (ELISA). Western blot analysis using the four MAbs showed specific recognition of AIB1 protein as a 160 kDa band in lysates from cell lines containing AIB1 gene-amplification. The MAbs immunoprecipitated in vitro-translated AIB1 and cellular AIB1 from metabolically labeled cells. The results show that these newly described MAbs are useful in studies of AIB1 function and expression. PMID- 10475244 TI - Monoclonal antibodies to progesterone: characterization and selection for enzyme immunoassay in bovine milk. AB - Thirty-one stabile murine monoclonal antibody (MAb) producing cell lines to progesterone were generated by using a short and a long immunization protocol. Long-term immunization with high doses of 11alpha-hydroxyprogesterone hemisuccinate-bovine serum albumin (11alpha-OH-P-HS-BSA) antigen led to very good antibody response in Balb/c mice. The donor mouse produced antiserum with a high titre of 1/250,000. Eleven MAbs were selected for further characterization since they showed high sensitivities (<35 pg/well to inhibit 50% of the tracer) in bridge homologous enzyme immunoassay (EIA). The results were compared to the donor mouse polyclonal antiserum. The MAbs and the donor mouse antiserum were generally found to be highly specific, when tested with 30 different steroids. Employing MAb 9C11, with affinity constant, K(alpha), to 11alpha-OH-P-HS of 1.1 x 10(10) M(-1), a bridge heterologous microtitre plate EIA for milk progesterone was developed, using the second-antibody coating technique and horseradish peroxidase (HRP) as an enzyme label. The assay is simple and convenient to use, as it permits direct addition of undiluted milk samples, at the same time maintaining high sensitivity, high precision, and a wide range of optical density (OD) values. The major advantage of the assay developed, compared to previously published direct addition milk progesterone immunoassays, is that progesterone concentrations, measured by the EIA, were not influenced by changing milk fat concentrations, even when milk samples containing up to 10% of milk fat were used for analysis. PMID- 10475245 TI - Primer mixture enhances PCR detection of Mycoplasma/Acholeplasma contaminants in cell cultures. PMID- 10475246 TI - Fetal thymic organ culture in rotating bioreactors. PMID- 10475248 TI - An embryonic cell line from the Caribbean fruit fly, Anastrepha suspensa (Diptera: Tephritidae) PMID- 10475247 TI - Differential response of the tail and body epidermis of Rana catesbeiana tadpoles in vitro to the anticancer drug, cisplatin. PMID- 10475249 TI - Differential response of embryonic stem cells and teratocarcinoma cells to carnosine. PMID- 10475250 TI - UV-induced but P53 independent apoptotic death in CHO.K1 cells is promoted by M phase inhibitors. PMID- 10475251 TI - Influence of antiestrogens on EGF- and IGF-I-mediated proliferation of human breast cancer cells. PMID- 10475252 TI - Behavior of a cell line derived from normal human hepatocytes on non physiological and physiological-type substrates: evidence for enhancement of secretion of liver-specific proteins by a three-dimensional growth pattern. AB - The behavior of a recently described cell line, HH25, derived from normal human hepatocytes, has been investigated on several different substrates--tissue culture plastic, glass, a thin layer of rat-tail collagen I, and thin layers or thick gels of extracellular matrix derived from the Engelbreth-Holm-Swarm murine sarcoma (EHS matrix). Cellular morphology, proliferation, and secretion of three hepatocyte-specific proteins (albumin, alpha1 acid glycoprotein, and alpha1 antitrypsin) have been examined. There were no differences in morphology, proliferation, or differentiated function in the cells on either plastic, glass, collagen, I, or a thin layer of EHS matrix, but on a thick EHS matrix gel the cells altered their morphology (forming three-dimensional colonies with canalicular-like structures) and their production of albumin and alpha1 acid glycoprotein was enhanced. This suggests that the enhanced differentiated function is associated with the morphological change (occurring only on the thick EHS gel) rather than with receptor-mediated cell-matrix interactions (which can also occur on the thin layer of EHS matrix). This cell line is therefore a good in vitro cellular model for the investigation of the roles of morphological changes and of cell-cell and cell-matrix interactions in the control of human hepatocyte behavior without the need for an extensive source of primary tissue. PMID- 10475253 TI - Characterization of the rat adrenal medulla cultured in vitro. AB - A wide variety of experimental animal models have been used to investigate the mechanisms of synthesis, storage, and release of catecholamines. Whereas in vivo experimental models are situated at one end of the spectrum, cell culture models are situated at the other end. In the present study, we have characterized various aspects of the rat adrenal medulla cultured in vitro as a whole tissue, aiming to establish a new experimental model in between in vivo animal models and cell culture models. We adapted a bottle rotator system commonly used for culturing rodent whole embryos. Changes in histology, activities and mRNA levels of catecholamine-synthesizing enzymes, and concentrations of catecholamines in the adrenal medulla were studied. In addition, the effects of cholinergic stimulation on catecholamine release from the adrenal medulla were examined. Overall the results indicate that various aspects of the adrenal medulla become stable after 4 d of culture and the adrenal medulla at this stage releases catecholamines in response to cholinergic stimulation. The whole adrenal medulla culture system may be a useful tool for investigating catecholamine-related functions dependent on intercellular reactions or communications. PMID- 10475254 TI - Lipopolysaccharide of Escherichia coli, polyamines, and acetic acid stimulate cell proliferation in intestinal epithelial cells. AB - Our aim was to examine whether lipopolysaccharide of Escherichia coli, polyamines of dietetic and/or bacterial origin, and products of the bacterial metabolism influence cell proliferation in epithelial cells from the colon and small intestine. Lipopolysaccharide of Escherichia coli 0111:B4 was incubated with cultures from human colonic mucosa. The mitoses were arrested with Vincristine and the total number of metaphases per crypt was counted. In addition, lipopolysaccharide was incubated with a human colonic epithelial cell line from adenocarcinoma (LS-123 cells) and with a nontransformed small intestinal cell line from germ-free rats (IEC-6 cells) for 24 h. In the last 4 h, the cells were labeled with tritiated thymidine. The cells were incubated with putrescine, cadaverine, and spermidine at 10(-11)-10(-3) M and with acetic acid (10(-5)-10( 1) M), acetaldehyde (10(-10)-10(-4) M) and ammonium chloride (1-20 mM). Lipopolysaccharide of Escherichia coli increased the number of arrested metaphases in human colonic crypts and DNA synthesis in L-123 and IEC-6 cells (P < 0.001). All polyamines increased DNA synthesis in the colonic and small intestinal cell lines, the effects being more marked for putrescine (P < 0.001). The higher concentrations of acetic acid increased DNA synthesis in both epithelial cell lines (P < 0.001). Acetaldehyde slightly decreased DNA synthesis in LS-123 cells at cytotoxic concentrations. Ammonium chloride did not significantly affect DNA synthesis. The final concentration of nonionized ammonia was less than 3%. It is concluded that lipopolysaccharides of Escherichia coli and intraluminal factors derived from microorganisms increase cell proliferation in human colonic crypts and intestinal epithelial cell lines. PMID- 10475255 TI - Suppression of Epstein-Barr virus reactivation in lymphoblastoid cells cultured in simulated microgravity. AB - Rotating-wall vessels allow for the growth of cells in simulated microgravity. Lymphoblastoid cells cultured in rotating-wall vessels exhibited significant differences in the expression of both early and late Epstein-Barr Virus (EBV) antigens. Viral protein expression (as measured by indirect immunofluorescence) was significantly suppressed in cells cultured in simulated microgravity. A significantly greater percentage of P3HR-1 cells and Daudi cells were positive for the expression of BamH1-Z-DNA fragment of Epstein-Barr replication activator (ZEBRA), early antigen restricted (EA-R), and viral capsid antigen (VCA) in cells cultured in static tissue culture flasks as compared to cells cultured in rotating-wall vessels. We observed a 7, 11, and 25-fold reduction, respectively, for EA-R, VCA, and ZEBRA protein in P3HR-1 cells cultured in simulated microgravity. Additionally, suspension cultures of P3HR-1 cells exhibited significantly greater ZEBRA antigen expression than cells cultured in rotating wall vessels. As an independent confirmation of the reduction in ZEBRA-protein production in simulated microgravity in P3HR-1 cells, ZEBRA-mRNA was quantitated by reverse transcription polymerase chain reaction. We observed between a 4 to 10 fold reduction in ZEBRA-mRNA in cells cultured in simulated microgravity as compared to cells cultured at 1 x g in tissue culture flasks. Rotating-wall vessels, by virtue of providing a simple culture environment triggering marked differences in viral activation, provide a model whereby both host and viral factors involved in regulating the maintenance of EBV latency can be examined. PMID- 10475256 TI - Improved soft-agar colony assay in a fluid processing apparatus. AB - The standard method for quantitating bone marrow precursor cells has been to count the number of colony-forming units that form in semisolid (0.3%) agar. Recently we adapted this assay for use in hardware, the Fluid Processing Apparatus, that is flown in standard payload lockers of the space shuttle. When mouse or rat macrophage colony-forming units were measured with this hardware in ground-based assays, we found significantly more colony growth than that seen in standard plate assays. The improved growth correlates with increased agar thickness but also appears to be due to properties inherent to the Fluid Processing Apparatus. This paper describes an improved method for determining bone marrow macrophage precursor numbers in semisolid agar. PMID- 10475257 TI - The modulation of cell shape influences porcine preadipocyte differentiation. PMID- 10475258 TI - Cryopreserved normal macrophages as a control for assays of macrophage function. PMID- 10475259 TI - Further optimization of culture method for rat keratinocytes: titration of glucose and sodium chloride. AB - The present study further improved the serum-free method of culturing rat keratinocytes. To obtain the best growth of rat keratinocytes, we modified our previous serum-free medium (MCDB153 based medium), particularly the amounts of glucose and sodium chloride (NaCl). Titration experiments showed the optimal concentration to be 0.8 mM for glucose and 100 mM for NaCl. This modification eliminated the requirement for albumin, which had been essential for colony formation when our previous medium was used. Titration of glucose and NaCl, followed by adjustment of essential amino acids and growth factors, produced a new formulation. More satisfactory and better growth was achieved with the new medium than with the previous medium. Accumulation of monoalkyldiacylglycerol (MADAG) was consistently noted in this study, representing the unusual lipid profile. A tendency toward normalization was, however, noted with the neutral lipid profile of keratinocytes cultivated in the new medium: lower production of MADAG was obtained with the new formulation, rather than the previous one. PMID- 10475260 TI - Method to analyze collagenase and gelatinase activity by fibroblasts in culture. AB - The net amount of collagen produced and deposited by fibroblasts in cell culture is determined by the rate of collagen synthesis as well as the rate of collagen degradation. Although collagen synthesis can be analyzed by several techniques, it is more difficult to measure collagen degradation. Breakdown of collagen depends upon the activity of a family of structurally and catalytically related mammalian enzymes termed matrix metalloproteinases (MMPs). Interstitial collagenase (MMP1) initiates the cleavage of fibrillar collagen, whereas gelatinases (MMP2 and MMP9) digest the denatured collagen fragments. A method has been developed to quantitate the activity of collagenase (MMP1) and gelatinase (MMP9) in conditioned medium from fibroblast cell cultures. The assay, which uses the fluorogenic substrate Dnp-Pro-Cha-Gly-Cys(Me)-His-AlaLys(Nma)NH2, is technically simple and amenable to high throughput analysis. Addition of specific inhibitors of the metalloproteinases allows for simultaneous measurement of both collagenase and gelatinase activity. PMID- 10475261 TI - A stromal cell line from rainbow trout spleen, RTS34ST, that supports the growth of rainbow trout macrophages and produces conditioned medium with mitogenic effects on leukocytes. AB - A rainbow trout spleen cell line, RTS34, was developed from a long-term hemopoietic culture. This cell line consisted of a mixed stromal cell layer with an associated cell population of macrophage-like cells that formed proliferative foci and released nonadherent progeny cells into the culture medium. A stromal cell line, RTS34st, was isolated from the RTS34 cell line. RTS34st cultures contained cells with fibroblast-like and epithelial-like morphologies and showed enhanced [3H]thymidine incorporation in response to either FBS or rainbow trout serum. The combination of FBS and trout serum was synergistic. Conditioned medium from RTS34st stimulated thymidine incorporation by peripheral blood and head kidney leukocytes, but not by leukocytes from the spleen. In addition, RTS34st provided a hemopoietic inductive microenvironment for immature precursor cells, selectively supporting the growth of macrophage-like cells. Therefore, RTS34st appears useful for studying the different roles of the stroma in regulating hemopoiesis in fish. PMID- 10475262 TI - Bio-stretch, a computerized cell strain apparatus for three-dimensional organotypic cultures. AB - In the present study, a unique mechanical strain apparatus for three-dimensional organotypic cultures was developed into a computerized system. It consists of a personal computer running Windows-based software, the Bio-Stretch Manager, a Bio Stretch Controller, and three sets of magnet boards. Cells are cultured on a Gelfoam sponge that is placed in a 35 mm petri dish with one end glued to the dish, and the other end attached to a coated steel bar. The petri dish is placed in front of a magnet, and the movement of the steel bar is controlled by dynamically changing the magnetic field. Up to five stretch patterns of variable frequency, duty cycle, and magnitude can be designed for each stretch regimen. Three different stretch regimens can be tested simultaneously. The operational characteristics of sponges were examined. Attachment of cells to the sponges was observed on several cell types. These features provide wide options for using this system to study the effects of mechanical stretch on cells. PMID- 10475263 TI - The establishment of heliothine cell lines and their susceptibility to two baculoviruses. AB - A total of eight cell lines were established from Helicoverpa armigera (3) and H. punctigera (5) embryos and ovaries. Cell lines were established and grown in TC100 and/or TC199-MK containing 10% fetal bovine serum. The serum-free medlium ExCell 400 was also used, with and without 10% supplemental fetal bovine serum, but failed to generate cell lines from fat bodies, embryos, or ovarian tissues. Cell lines consisted of heterogenous cell types ranging from oval to fibroblast like. This is the first report on the successful establishment of cell lines from H. punctigera. Cell lines from the two species were distinguishable from each other by DAF-PCR, and noticeable differences in minor bands were observed among cell lines from the same species. All of the established cell lines from both species were susceptible to HzSNPV but did not replicate more virus than that of a H. zea cell line (BCIRL-HZ-AM1-A11). However, an H. punctigera cell line (HP1) replicated AcMNPV to the highest titer (1.0 X 10(8) 50% tissue culture infective dose/ml), and only one of the H. armigerm cell lines (HA1) was susceptible to this virus. PMID- 10475264 TI - Exposure to hyperbaric oxygen induces cell cycle perturbation in prostate cancer cells. AB - Cell cycle synchronization of tumor cells by exposure to hyperbaric oxygenation (HBO) may increase the efficacy of chemotherapy or radiation by placing cells into a chemosensitive portion of the cycle. The purpose of the current study was to examine oxygen pressure-dependent relationships with respect to the cell cycle in prostate tumor cells in vitro. LNCaP cells were grown in an incubator at 21% O2 and then exposed to 100% oxygen at pressures up to 6 atmospheres (atm) for 1.5 h. Cells were then returned to the incubator and evaluated for DNA content by propidium iodide and new DNA synthesis with a pulse-chase experiment. Cell cycle effects were evaluated by flow cytometry. Exposure to HBO increased the percentage of cells synthesizing new DNA in a dose-dependent fashion: 0 atm, 44%; 6 atm, 65%. Cells that synthesize new DNA accumulate in G2/M as a function of partial pressure of oxygen. These results suggest that HBO induces cells to enter the cell cycle and accumulate in G2/M. Cell cycle synchronization and entry of senescent cells into the cell cycle suggest that HBO may be a useful adjuvant to chemotherapy or radiation in the treatment of prostate cancer. There are two potential mechanisms of action that may make HBO efficacious in the treatment of prostate cancer. HBO may potentiate cancer chemotherapeutic agents that cause damage to DNA during DNA synthesis or HBO may inhibit cell division causing accumulation in G2/M. PMID- 10475266 TI - Transformation of lepidopteran and coleopteran insect cell lines by Glyptapanteles indiensis polydnavirus DNA. AB - Recently investigators showed that polydnavirus DNA from the parasitic wasp Glyptapanteles indiensis could transform gypsy moth L. dispar cell lines in vitro (McKelvey et al., 1996). Here we show GiPDV DNA is capable of transforming in vitro to varying degrees lepidopteran (IPLB-TN-R2, IPLB-SF-21, IAL-PID2, IPLB HvT1) and coleopteran (IPLB-DU182E) insect cell lines derived from various somatic tissue types. An insect cell line derived from dipteran Aedes albopictus (C7/10) could not be transformed with G. indiensis polydnavirus. PMID- 10475265 TI - In vitro analysis of venom from the wasp Nasonia vitripennis: susceptibility of different cell lines and venom-induced changes in plasma membrane permeability. AB - The lethal effects of crude venom prepared from the ectoparasitic wasp Nasonia vitripennis were examined with cultured cells from six insect and two vertebrate species. Venom caused cells from Sarcophaga peregrina (NIH SaPe4), Drosophila melanogaster (CRL 1963), Trichoplusia ni (TN-368 and BTI-TN-5B1-4), Spodoptera frugiperda (SF-21AE), and Lymantria dispar (IPL-Ldfbc1) to round up, swell, and eventually die. Despite similar sensitivities and overlapping LC50 values [0.0004 0.0015 venom reservoir equivalents (VRE)/microl], profound differences were noted at the onset of cytotoxicity among the six insect cell lines: over 80% of the NIH SaPe4 and SF21AE cells were nonviable within 1 h after addition of an LC99 dose of venom, whereas the other cells required a 5-10-fold longer incubation period to produce mortality approaching 100%. In contrast, cells from the grass frog, Rana pipiens (ICR-2A), and goldfish, Carassius auratus (CAR), showed little sensitivity to the venom: six venom reservoir equivalents were needed to induce 50% mortality in ICR-2A cells [50% lethal concentration (LC50) = 0.067 VRE/microl), and 9 VRE did not yield sufficient mortality in CAR cells for us to calculate an LC50. All susceptible cells showed similar responses when incubated with wasp venom: retraction of cytoplasmic extensions (when present), blebbing of the plasma membrane, swelling of the plasma and nuclear membranes, condensation of nuclear material, and eventual cell death attributed to lysis. The rate of swelling and lysis in NIH SaPe4 and BTI-TN-5B1-4 cells exposed to venom appeared to be dependent on the diffusion potential of extracellular solutes (Na+ = choline > sucrose > or = raffinose > K+), which is consistent with a colloid osmotic lysis mechanism of cell death. When T. ni cells were cotreated with venom and the K+ channel blocker 4-aminopyridine, cell swelling and lysis increased with increasing drug concentration. In contrast, cells from S. peregrina were protected from the effects of the venom when treated in a similar manner. Addition of certain divalent cations (Zn+2 and Ca+2) to the extracellular media 1 h postvenom incubation rescued both BTI-TN-5B1-4 and NIH SaPe4 cells, suggesting that protection was gained from closure of open pores rather than prevention of pore formation. Venom from N. vitripennis displayed no hemolytic activity toward sheep erythrocytes, supporting the view that venom intoxication is not by a nondiscriminate mechanism. A possible mode of action of the venom is discussed. PMID- 10475267 TI - Improved recovery of active radiolabeled TGFbeta1 by TGFbeta receptor type III affinity chromatography. PMID- 10475268 TI - Authentication of two cell lines developed from the larval and pupal ovaries of Spodoptera litura by rRNA based methods. PMID- 10475269 TI - SV40 large T antigen immortalization of rat hepatic stellate-like cells. PMID- 10475270 TI - Comparison of cell line maintenance procedures on insect cells used for producing baculoviruses. AB - A gypsy moth cell line, IPLB-LdEIta, maintained under various conditions was tested for susceptibility to and productivity of two baculoviruses, the Autographa californica nucleopolyhedrovirus (AcMNPV) and Lymantria dispar nucleopolyhedrovirus (LdMNPV). The results suggest that cells maintained in serum containing medium (modified TC100) were more susceptible (on the basis of titers in an endpoint assay) to LdMNPV than cells maintained in a serum-free medium (ExCell 400). Such a difference was not apparent with AcMNPV. Similarly, little difference existed in the proportion of cells containing occlusion bodies (OBs) a wk after inoculation with AcMNPV (i.e., the percent infected) in any LdEIta strains, although one combination of cells and medium (cells maintained in ExCell 400 but infected in TC100) showed a lower percent infection with LdMNPV. Even though the percentage of cells infected varied little, the number of OBs produced varied by 3 logs with AcMNPV and 11/2 logs with LdMNPV. In each case, cells normally grown in ExCell 400 and infected in the same medium produced the lowest number of OBs. However, productivity was improved when cells normally grown in ExCell 400 were infected in TC100. Even more interesting was that cells normally grown in TC100 produced more AcMNPV OBs when infected in ExCell 400 medium. This suggests that changing culture medium (regardless of the normal maintenance medium) can stimulate virus production. In addition to examining virus productivity in LdEIta cells in both serum-containing and serum-free media, I also tested a strain maintained at low temperature (17 degrees C) for over a yr. This maintenance protocol was not detrimental for LdMNPV productivity and was slightly stimulatory for production of AcMNPV. PMID- 10475271 TI - A protein from Naegleria amoebae causes apoptosis in chick embryo and CHO cells after they become confluent. AB - Exposure for less than an hour to a protein isolated from Naegleria amoebae initiates a process that has no apparent effect on the appearance or growth of chick embryo or CHO cell cultures for 4 to 9 days; after the development of confluency, at some unknown signal, all of the cells undergo an apoptotic death within a 12- to 24-hour period. Abnormalities detected among the last mitotic cells include chromosomal breakage and early reversal in metaphase to telo/interphase daughter nuclei with irregular shapes. Additional events in the dying cultures include the development of a cytoplasmic amoebic-related immunogen, gross DNA fragmentation, cell blebbing, shrinkage, and apoptotic body formation. Culture death included all cells, those present in confluent cultures when the protein was added, and in other cultures, those formed during a more than 30-fold increase in cells as the cultures became confluent. The increase in the number of cells followed by the uniformity and synchrony of their death pattern indicates that the signal to kill has increased and spread throughout the culture; upon an unknown condition related to confluency, events are initiated that lead to the unusual apoptotic death of the culture. PMID- 10475272 TI - High density micromass cultures of embryonic limb bud mesenchymal cells: an in vitro model of endochondral skeletal development. AB - To study the mechanisms regulating endochondral skeletal development, we examined the characteristics of long-term, high density micromass cultures of embryonic chicken limb bud mesenchymal cells. By culture Day 3, these cells underwent distinct chondrogenesis, evidenced by cellular condensation to form large nodules exhibiting cartilage-like morphology and extracellular matrix. By Day 14, extensive cellular hypertrophy was seen in the core of the nodules, accompanied by increased alkaline phosphatase activity, and the limitation of cellular proliferation to the periphery of the nodules and to internodular areas. By Day 14, matrix calcification was detected by alizarin red staining, and calcium incorporation increased as a function of culture time up to 2 to 3 wk and then decreased. X-ray probe elemental analysis detected the presence of hydroxyapatite. Analogous to growth cartilage developing in vivo, these cultures also exhibited time-dependent apoptosis, on the basis of DNA fragmentation detected in situ by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL), ultrastructural nuclear morphology, and the appearance of internucleosomal DNA degradation. These findings showed that cellular differentiation, maturation, hypertrophy, calcification, and apoptosis occurred sequentially in the embryonic limb mesenchyme micromass cultures and indicate their utility as a convenient in vitro model to investigate the regulatory mechanisms of endochondral ossification. PMID- 10475273 TI - Parabolic growth patterns in 96-well plate cell growth experiments. AB - In preparing for the routine use of the ubiquitous in vitro cell growth inhibition assay for the study of anticancer agents, we characterized the statistical properties of the assay and found some surprising results. Parabolic well-to-well cell growth patterns were discovered, which could profoundly affect the results of routine growth inhibition studies of anticancer and other agents. Four human ovarian cell lines, A2780/WT, A2780/DX5, A2780/DX5B, and A121, and one human ileocecal adenocarcinoma cell line, HCT-8, were seeded into plastic 96-well plates with a 12-channel pipette, without drugs, and grown from 1-5 d. The wells were washed with a plate washer, cells stained with sulforhodamine B (SRB), and dye absorbance measured with a plate reader. Variance models were fit to the data from replicates to determine the nature of the heteroscedastic error structure. Exponential growth models were fit to data to estimate doubling times for each cell line. Polynomial models were fit to data from 10-plate stacks of 96-well plates to explore nonuniformity of cell growth in wells in different regions of the stacks. Each separate step in the assay was examined for precision, patterns, and underlying causes of variation. Differential evaporation of water from wells is likely a major, but not exclusive, contributor to the systematic well-to-well cell growth patterns. Because the fundamental underlying causes of the parabolic growth patterns were not conclusively found, a randomization step for the growth assay was developed. PMID- 10475275 TI - Primary cultures of heart cells from the scallop Pecten maximus (mollusca bivalvia). AB - Primary cultures of Pecten maximus heart cells, isolated by an enzymatic procedure, were routinely obtained with a high level of reproducibility in a simple medium based on sterile seawater. Cells attached to the plastic substratum without the need to add a special factor. The number of adhering cells gradually increased with the time of culture. Two types of adhering cells were observed: epitheliallike cells and fibroblastlike cells, which were more numerous. The latter cells were identified as myocytes by electron microscopy and immunofluorescent staining. Results obtained by autoradiography, after incorporation of [14C]leucine, [3H]thymidine, and [14C]acetate, confirmed functional activity of the cells. These cultures were maintained viable in vitro during at least 1 mo. PMID- 10475274 TI - A novel human astrocyte cell line (A735) with astrocyte-specific neurotransmitter function. AB - Studies of brain cell function and physiology are hampered by the limited availability of immortal human brain-derived cell lines, as a result of the technical difficulties encountered in establishing immortal human cells in culture. In this study, we demonstrate the application of recombinant DNA vectors expressing SV40 T antigen for the development of immortal human cell cultures, with morphological, growth, and functional properties of astrocytes. Primary human astrocytes were transfected with the SV40 T antigen expression vectors, pSV3neo or p735.6, and cultures were established with an extended lifespan. One of these cultures gave rise to an immortal cell line, designated A735. All the human SV40-derived lines retained morphological features and growth properties of type 1 astrocytes. Immunohistochemical studies and Western blot analysis of the intermediate filament proteins and glutamine synthetase demonstrated a differentiated but immature astrocyte phenotype. Transport of gamma-amino butyric acid and glutamate were examined and found to be by a glial-specific mechanism, consistent with the cell lines' retaining aspects of normal glial function. We conclude that methods based on the use of SV40 T antigen can successfully immortalize human astrocytes, retaining key astrocyte functions, but T antigen induced proliferation appeared to interfere with expression of glial fibrillary acidic protein. We believe A735 is the first documented nontumor-derived human glial cell line which is immortal. PMID- 10475276 TI - 1,25-dihydroxyvitamin D3 inhibits parathyroid hormone-related peptide mRNA expression in fetal rat long bones in culture. AB - When fetal rat long bones are incubated in the presence of 10(-8) M 1,25 dihydroxyvitamin D3 [1,25(OH)2D3], steady-state parathyroid hormone-related peptide (PTHrP) mRNA levels are decreased. This decrease is temporary: it is observed as soon as after 3 h of exposure and reaches a nadir after 6 h. At that time, PTHrP mRNA levels are significantly lower in the experimental than in the control bones. However the inhibitory effect vanishes after 24 h, despite continuous exposure to 1,25(OH)2D3 for even 48 h. This is the first report showing that PTHrP mRNA expression can be regulated in rat fetal long bones in vitro by 1,25(OH)2D3. PMID- 10475277 TI - Activity spectra of Bacillus thuringiensis delta-endotoxins against eight insect cell lines. AB - Eight continuous insect cell lines were tested for susceptibility to the delta endotoxins of several lepidopteran-active strains and cloned-gene products of Bacillus thuringiensis. The assays were performed on cells suspended in agarose gel, which allowed the toxins activated at pH 10.5 to be applied directly in a high-pH buffer without causing solvent toxicity to the cells. The responses of the cell lines to the various toxins produced activity spectra that were used to identify functionally similar and dissimilar toxin proteins. IPRI-CF-1 and FPMI MS-5, derived from neonate larvae of Choristoneura fumiferana and Manduca sexta, respectively, exhibited the greatest sensitivity to the toxins tested, whereas B. thuringiensis subsp. entomocidus had the broadest in vitro host range. Analysis of activity spectra led to the identification of the particular Cry protein that was responsible for the broad toxicity of this subspecies and demonstrated a distinct difference in toxin composition between two strains of subsp. sotto. The identical spectra observed for subsp. kurstaki HD-1 and NRD-12 is consistent with insect bioassay data obtained previously by other workers and supports the conclusion that there is virtually no difference in activity between these two strains. The in vitro assay system, referred to as the "lawn assay" and used to test B. thuringiensis activated toxins against insect cell lines, is particularly useful in mode-of-action studies and as a rapid, preliminary test for the presence of specific cytolytic proteins, rather than as a method for screening toxins of wild-type strains for insecticidal activity. The response of cells in vitro to B. thuringiensis toxins is often very different from that of the insect from which the cells were derived. PMID- 10475278 TI - What determines anesthetic induction dose? It's the front-end kinetics, doctor! PMID- 10475279 TI - Cardiac output is a determinant of the initial concentrations of propofol after short-infusion administration. AB - Indicator dilution theory predicts that the first-pass pulmonary and systemic arterial concentrations of a drug will be inversely related to the cardiac output. For high-clearance drugs, these first-pass concentrations may contribute significantly to the measured arterial concentrations, which would therefore also be inversely related to cardiac output. We examined the cardiac output dependence of the initial kinetics of propofol in two separate studies using chronically instrumented sheep in which propofol (100 mg) was infused IV over 2 min. In the first study, steady-state periods of low, medium, and high cardiac output were achieved by altering carbon dioxide tension in six halothane-anesthetized sheep. The initial area under the curve and peak value of the pulmonary artery propofol concentrations were inversely related to cardiac output (R2 = 0.57 and 0.66, respectively). For the systemic arterial concentrations, these R2 values were 0.68 and 0.71, respectively. In our second study, transient reductions in cardiac output were achieved in five conscious sheep by administering a short infusion of metaraminol concurrently with propofol. Cardiac output was lowered by 2.2 L/min, and the area under the curve to 10 min of the arterial concentrations increased to 143% of control. IMPLICATIONS: The initial arterial concentrations of propofol after IV administration were shown to be inversely related to cardiac output. This implies that cardiac output may be a determinant of the induction of anesthesia with propofol. PMID- 10475280 TI - Adverse cardiac outcomes after noncardiac surgery in patients with prior percutaneous transluminal coronary angioplasty. AB - In this retrospective cohort study, we compared adverse cardiac outcomes after noncardiac surgery among patients with prior percutaneous transluminal coronary angioplasty (PTCA), patients with nonrevascularized coronary artery disease (CAD), and normal controls. Inpatient hospital discharge abstracts from all nonfederal acute care hospitals in Washington State linked to death certificates were evaluated. Patients > or =45 yr old with prior PTCA who underwent noncardiac surgery from 1987 to 1993 were matched by age, sex, surgery type, and discharge year to 686 patients with CAD and to 2155 normal controls (no CAD). We compared risk for adverse cardiac outcomes (death, myocardial infarction, angina, congestive heart failure, malignant dysrhythmia, cardiogenic shock, coronary artery bypass graft, or PTCA) within 30 days. Patients with PTCA had twice the risk of adverse cardiac outcome as normal controls (odds ratio [OR] 1.98; P < 0.001), with a higher risk of angina (OR 7.84), congestive heart failure (OR 2.06), and myocardial infarction (OR 3.86) but a lower risk of death (OR 0.46; P < 0.001). Patients with PTCA had half the risk of adverse cardiac outcome as patients with CAD (OR 0.50; P < 0.001), including less risk of angina (OR 0.51) and congestive heart failure (OR 0.40; P < 0.001), but no difference in myocardial infarction (P = 0.304) or death (P = 0.436). No difference was found between 142 patients with recent PTCA (< or =90 days before noncardiac surgery) matched to patients with CAD (OR 0.90; P = 0.396). Patients revascularized by PTCA >90 days before noncardiac surgery seem to have a lower risk of poor outcome than nonrevascularized patients, although not as low as normal controls. For recent PTCA patients, the lack of difference compared with CAD patient outcomes requires a larger sample size for verification. Present findings do not lend support to a role for prophylactic PTCA to improve noncardiac surgery outcomes. This investigation did not control for CAD severity, medical management, or comorbidities. Study of these factors is needed before the clinical implications of PTCA for noncardiac surgical risk can be completely assessed. IMPLICATIONS: Hospital records showed patients with prior percutaneous transluminal coronary angioplasty were twice as likely as healthy patients to have an adverse cardiac outcome after noncardiac surgery, although their risk was reduced by half compared with patients with untreated coronary artery disease. Further study of the role of percutaneous transluminal coronary angioplasty in modulating noncardiac surgery risk is needed. PMID- 10475281 TI - Isoflurane administration before ischemia and during reperfusion attenuates ischemia/reperfusion-induced injury of isolated rabbit lungs. AB - To investigate the effects of isoflurane on ischemia/ reperfusion (IR)-induced lung injury, we administered isoflurane before ischemia or during reperfusion. Isolated rabbit lungs were divided into the following groups: control (n = 6), perfused and ventilated for 120 min without ischemia; ISO-control (n = 6), 1 minimum alveolar anesthetic concentration (MAC) isoflurane was administered for 30 min before 120 min continuous perfusion; IR (n = 6), ischemia for 60 min, followed by 60 min reperfusion; IR-ISO1 and IR-ISO2, ischemia followed by reperfusion and 1 MAC (n = 6) or 2 MAC (n = 6) isoflurane for 60 min; ISO-IR (n = 6), 1 MAC isoflurane was administered for 30 min before ischemia, followed by IR. During these maneuvers, we measured total pulmonary vascular resistance (Rt), coefficient of filtration (Kfc), and lung wet to dry ratio (W/D). The results indicated that administration of isoflurane during reperfusion inhibited an IR induced increase in Kfc and W/D ratio. Furthermore, isoflurane at 2 MAC, but not 1 MAC, significantly inhibited an IR-induced increase in Rt. The administration of isoflurane before ischemia significantly attenuated the increase in IR-induced Kfc, W/D, and Rt. IMPLICATIONS: Our results suggest that the administration of isoflurane before ischemia and during reperfusion protects against ischemia reperfusion-induced injury in isolated rabbit lungs. PMID- 10475282 TI - Aortic and radial pulse contour: different effects of nitroglycerin and prostacyclin. AB - Through vasorelaxation, nitroglycerin is considered to reduce arterial wave reflection and to cause a more pronounced decrease in systolic pressure in the aorta (AoSAP) than in the radial artery (RaSAP). Our aim was to study how radial and aortic pulse wave configurations and the gradient (RaSAP-AoSAP) were affected by nitroglycerin and by prostacyclin, and how these changes correlated to stroke volume, vascular resistance/impedance, and wave reflection. Prostacyclin has not been studied in this context and was chosen because, in contrast to nitroglycerin, it does not reduce stroke volume and reduces afterload by arteriolar dilation. In 18 patients (53-81 yr old; heavily premedicated before coronary artery surgery), blood pressure was measured in both the radial artery and the ascending aorta (tipmanometry), and cardiac output was measured by thermodilution. Mean arterial pressure was reduced stepwise with each drug (mean total decrease 10-12 mm Hg). The initial RaSAP-AoSAP gradient (6 mm Hg) was increased 10 mm Hg by nitroglycerin and was not affected by prostacyclin. The nitroglycerin-induced increase in systolic gradient RaSAP-AoSAP correlated to decreases in stroke volume index, mean arterial pressure, and arterial elastance, but not to decrease in pulse wave augmentation. Thus, decreases in stroke volume index, not wave reflection, seem to be the main reason for an increased RaSAP AoSAP when nitroglycerin is used in the elderly, hypertensive patient. IMPLICATIONS: We studied ascending aortic and radial pulse contours in patients scheduled for coronary artery surgery. The radial pulse wave can be used for interpretation of central hemodynamic changes during nitroglycerin-, but not prostacyclin-, induced hypotension. PMID- 10475283 TI - The effects of heparin coating of oxygenator fibers on platelet adhesion and protein adsorption. AB - Platelet adhesion on the cardiopulmonary bypass oxygenator membrane is associated with impaired hemostasis. We investigated the effects of heparin coating of the oxygenator membrane on protein adsorption and platelet adhesion on the surface. Noncoated and heparin-coated polypropylene membranes were incubated in whole blood with small- (1 U/mL) or large-dose (5 U/mL) heparin as an anticoagulant for 3 h at 37 degrees C. The amount of platelets adhering on each fiber was assessed by using enzyme immunoassays using monoclonal antibodies directed against CD42b (GP Ib) and CD61 (GP IIb/IIIa). Platelet activation was assessed by measuring plasma guanosine monophosphate 140 levels. The amount and composition of the adsorbed proteins on the surface were analyzed by using a bicinchoninic acid protein assay and by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting technique. The heparin coating of the fibers significantly reduced platelet adhesion on the surface. However, platelet activation was reduced by heparin coating only with small-dose heparinization. The adsorption of platelet adhesive proteins such as fibrinogen and von Willebrand factor was not altered, whereas that of fibronectin was increased by heparin coating. We conclude that heparin coating of the oxygenator fibers can decrease platelet adhesion without affecting adsorption of major adhesive proteins. Surface heparin coating is associated with an increased fibronectin adsorption on the fibers. IMPLICATIONS: Heparin coating can reduce platelet adhesion and activation in the presence of small-dose heparinization, potentially reducing the inflammatory response and activation of thrombosis and fibrinolysis. PMID- 10475284 TI - Use of abciximab-modified thrombelastography in patients undergoing cardiac surgery. AB - Thrombelastography (TEG) is a reliable coagulation monitoring system that can guide blood product transfusion in cardiac surgery. The maximum amplitude (MA) of TEG measures clot strength, which is dependent on both fibrinogen level and platelet function. Inhibition of platelet function with abciximab-fab is suggested to permit quantitative assessment of the contribution of fibrinogen to clot strength. We hypothesized that abciximab-modified TEG permits prediction of plasma fibrinogen levels and that the difference of standard MA and abciximab modified MA (deltaMA) is a correlate for platelet function. We correlated abciximab-modified MA with plasma fibrinogen levels and deltaMA with platelet count in patients undergoing coronary revascularization. Correlation between plasma fibrinogen levels and abciximab-modified MA was significant (adjusted r2: 0.8; P < 0.0001). Correlation of deltaMA with platelet count was not significant when calculated in millimeters (adjusted r2: 0.04; P = 0.73). However, when deltaMA was calculated in dynes per square centimeter (deltaGMA), it correlated significantly with platelet count (adjusted r2: 0.51; P < 0.0001). We conclude that abciximab-modified TEG may therefore help to discriminate between hypofibrinogenemia and platelet dysfunction as a cause of decreased MA. IMPLICATIONS: We examined the use of abciximab-modified thrombelastography in patients undergoing cardiac surgery. Modification of thrombelastography with abciximab-fab allows prediction of fibrinogen levels, despite coagulation altered by cardiac surgery. The difference of standard maximum amplitude and abciximab modified maximum amplitude correlates with platelet function when expressed in dynes per square centimeter. PMID- 10475285 TI - An assessment of the safety of short-term amiodarone therapy in cardiac surgical patients with fentanyl-isoflurane anesthesia. AB - In previously published case reports and a retrospective study, investigators have noted that amiodarone may cause substantial hemodynamic instability when combined with fentanyl-containing anesthesia regimens. We performed the present study to evaluate the safety of short-term amiodarone therapy when combined with a fentanyl-containing anesthesia regimen in a randomized, double-blinded, placebo controlled format. After institutional approval and written informed consent, patients scheduled to undergo coronary artery bypass grafting or valvular surgery were randomly allocated to receive amiodarone (3.4 g over 5 days or 2.2 g over 24 hours) or placebo before surgery. Four indicators for hemodynamic instability were assessed: 1) a net increase in fluid balance during surgery of >2 L; 2) use of dopamine at a rate >10 microg x kg(-1) x min(-1); 3) use of other vasopressive catecholamines; and 4) use of a phosphodiesterase inhibitor or intraaortic balloon pump. Systolic, diastolic, and central venous pressures were measured before fentanyl administration, before cardiopulmonary bypass (CPB), and after separation from CPB. Overall, 84 patients (45 patients in the amiodarone group, 39 in the placebo group) were enrolled and completed the study. There were no significant differences between the two groups in any indicator for hemodynamic instability or the indicators of instability combined. After CPB, there was a significantly lower systolic blood pressure in the amiodarone group compared with the placebo group (112 +/- 12 vs 117 +/- 14 mm Hg; P = 0.049). However, there was a trend toward smaller IV fluid requirements during surgery in the amiodarone group compared with the placebo group (438 +/- 867 vs 907 +/- 1640 mL; P = 0.09). We found no increased risk of hemodynamic compromise after short-term amiodarone therapy among patients receiving a fentanyl-containing anesthesia regimen during open heart surgery. IMPLICATIONS: In previous retrospective studies and case reports, investigators have identified a possible risk of hemodynamic compromise when patients receiving chronic amiodarone therapy are given anesthesia regimens containing fentanyl. We performed a prospective, randomized, double-blinded study to evaluate the hemodynamic effects of short-term amiodarone therapy during fentanyl-isoflurane anesthesia for open heart surgery. No adverse hemodynamic effects of amiodarone were identified. PMID- 10475286 TI - Reduction of blood loss and transfusion requirement by aprotinin in posterior lumbar spine fusion. AB - Aprotinin reduces blood loss in many orthopedic procedures. In posterior lumbar spine fusion, blood loss results primarily from large vein bleeding and also occurs after the wound is closed. Seventy-two patients undergoing posterior lumbar spine fusion were randomly assigned to large-dose aprotinin therapy or placebo. All patients donated three units of packed red blood cells (RBCs) preoperatively. Postoperative blood loss was harvested from the surgical wound in patients undergoing two- and/or three-level fusion for reinfusion. The target hematocrit for RBC transfusion was 26% if tolerated. Total (intraoperative and 24 h postoperative) blood loss, transfusion requirements, and percentage of transfused patients per treatment group were significantly smaller in the aprotinin group than in the placebo group (1935 +/- 873 vs 2809 +/- 973 mL per patient [P = 0.007]; 42 vs 95 packed RBCs per group [P = 0.001]; 40% vs 81% per group [P = 0.02]). Hematological assessments showed an identically significant (a) intraoperative increase in both thrombin-antithrombin III complexes (TAT) and in activated factor XII (XIIa) and (b) decrease in activated factor VII (VIIa), indicating a similar significant effect on coagulation in patients of both groups (P = 0.9 for intergroup comparisons of postoperative VIIa, XIIa, and TAT). Intraoperative activation of fibrinolysis was significantly less pronounced in the aprotinin group than in the placebo group (P < 0.0001 for intergroup comparison of postoperative D-dimer levels). No adverse drug effects (circulatory disturbances, deep venous thrombosis, alteration of serum creatinine) were detected. Although administered intraoperatively, aprotinin treatment dramatically reduced intraoperative and 24-h postoperative blood loss and autologous transfusion requirements but did not change homologous transfusion in posterior lumbar spine fusion. IMPLICATIONS: In our study, aprotinin therapy significantly decreased autologous, but not homologous, transfusion requirements in posterior lumbar spine fusion. PMID- 10475287 TI - Fast-track cardiac anesthesia in patients with sickle cell abnormalities. AB - We conducted a retrospective review of 10 patients with sickle cell trait (SCT) and 30 patients (cohort control) without SCT undergoing first-time coronary artery bypass graft surgery with cardiopulmonary bypass. Demographic, perioperative management, and outcome data were collected. Both groups were matched according to age, weight, duration of surgery, and preoperative hemoglobin (Hb) concentration. Distribution of gender, medical conditions, pharmacological treatment, and preoperative left ventricular function were similar between the groups. The comparisons were analyzed in respect to postoperative blood loss and transfusion rates, as well as duration of intubation, intensive care unit, and hospital length of stay (LOS). All patients underwent fast-track cardiac anesthesia. A combination of cold crystalloid and blood cardioplegia was used. The lowest nasopharyngeal temperature was 33 degrees C. There were no episodes of significant hypoxemia, hypercarbia, or acidosis. None of the patients had sickling crisis during the perioperative period. The postoperative blood loss was 687 +/- 135 vs 585 +/-220 mL in the SCT and control groups, respectively. The trigger for blood transfusion during cardiopulmonary bypass was hematocrit <20% and Hb <75 g/L postoperatively. Three SCT patients (30%) and 10 control patients (33%) received a blood transfusion. Median extubation time was 4.0 vs 3.9 h; intensive care unit LOS was 27 vs 28 h; and hospital LOS was 6.0 vs 5.5 days in the SCT and control groups, respectively. There were no intraoperative deaths. One patient in the SCT group died from multiorgan failure 2 mo after surgery. IMPLICATIONS: Fast-track cardiac anesthesia can be used safely in patients with sickle cell trait undergoing first time coronary artery bypass graft surgery. Extubation time and intensive care unit and hospital length of stay are comparable to those of matched controls, and blood loss and transfusion requirements are not increased. A hematocrit of 20% seems to be a safe transfusion trigger during cardiopulmonary bypass in these patients. PMID- 10475288 TI - Propofol-induced alterations in myocardial beta-adrenoceptor binding and responsiveness. AB - Propofol (iv) depresses cardiovascular function in both humans and animals. However, the mechanism underlying this action has not been well described. The present study was designed to test the hypothesis that this effect of propofol results in part from an antagonism of adrenergic control of the heart. Experiments examined effects of propofol on: 1) [3H]CGP12177 (a beta-adrenoceptor antagonist) binding in rat myocardial membranes; and 2) the inotropic and chronotropic actions of isoproterenol in rat left atrial muscle and right atria, respectively. Propofol (25-200 microM) increased the apparent dissociation constant for [3H]CGP12177 without affecting binding site density. Similarly, 200 microM propofol increased the 50% effective concentration values for the dose dependent positive chronotropic and inotropic actions of isoproterenol in right and left atria, and depressed the maximum increase in spontaneous rate elicited by this beta-adrenoceptor agonist. Other experiments demonstrated that propofol does not alter muscarinic receptor binding as monitored using [3H]quinuclidi nylbenzilate. In conclusion, these results indicate that propofol can decrease cardiac beta-adrenoceptor responsiveness; however, the concentrations of propofol required suggest that this action contributes to the cardiovascular depression produced by this anesthetic only during large-dose bolus injection. IMPLICATIONS: Experiments in membranes and cardiac preparations isolated from rat heart demonstrate that relatively high concentrations of propofol (25-200 microM) are required to antagonize beta-adrenoceptor binding and tissue responsiveness. PMID- 10475289 TI - Wolff-Parkinson-White syndrome simulating inferior myocardial infarction in a cocaine abuser for urgent dilation and evacuation of the uterus. PMID- 10475290 TI - Anesthetic and perioperative management of adult transplant recipients in nontransplant surgery. PMID- 10475291 TI - Comparison of vital capacity induction with sevoflurane to intravenous induction with propofol for adult ambulatory anesthesia. AB - We compared vital capacity inhaled induction (VC) with sevoflurane with i.v. induction with propofol for adult ambulatory anesthesia. Patients were randomly assigned to receive either 8% sevoflurane in 75% N2O/O2 from a primed circuit (VC, 32 patients) or propofol 2-mg/kg bolus (i.v., 24 patients). Times to loss of consciousness (response to command) and induction side effects (airway, hemodynamic, motor) were assessed. Anesthesia was maintained with sevoflurane/N2O via a face mask for both groups. At the end of surgery, recovery times were measured and psychomotor function tests were performed. Patients were also asked to assess the quality of their anesthesia. Of the VC patients, 59% lost responsiveness in one breath, taking 39 +/- 3 s. All VC patients completed the induction, and all measures of induction time were significantly shorter for VC than for i.v. Induction side effects were different in the two groups (cough and hiccough for VC versus movement and blood pressure changes for i.v.), but overall incidences were similar. There were no significant differences in any index of early or intermediate recovery. Mild nausea occurred more often with VC, but no antiemetics were needed, and discharge was not delayed. Patients' assessments of the quality of induction or wake up were not significantly different between VC and i.v. Thus, VC induction with sevoflurane is an acceptable alternative to propofol i.v. induction of general anesthesia for adult ambulatory surgical patients. IMPLICATIONS: A vital capacity induction with sevoflurane produced a faster loss of consciousness and had side effects, recovery times, and patient satisfaction similar to that of a propofol induction in adults undergoing ambulatory surgery. PMID- 10475292 TI - The effects of antagonizing residual neuromuscular blockade by neostigmine and glycopyrrolate on nausea and vomiting after ambulatory surgery. AB - The effects of neostigmine on the incidence of postoperative nausea and vomiting (PONV) are controversial. In this study, we evaluated the effects of neostigmine and glycopyrrolate on the incidence of PONV and the need for antiemetics in patients undergoing ambulatory surgery. One hundred healthy patients undergoing outpatient surgical procedures were included in the study. A standardized anesthetic technique was used for all patients. Patients were randomized to receive either mivacurium (n = 50) or rocuronium (n = 50) to achieve muscle paralysis. Bolus doses of mivacurium 2-4 mg or rocuronium 5-10 mg were administered to maintain one or two twitches of the train-of-four stimulation of the ulnar nerve at the wrist. After surgery, residual neuromuscular blockade was reversed with neostigmine 2.5 mg i.v. and glycopyrrolate 0.5 mg i.v. only if clinically deemed necessary (i.e., fade on train-of-four stimulation, inadequate tidal volume, reduced hand grip, or inability to maintain head lift). The incidence of PONV and the need for antiemetics were recorded in the post anesthesia care unit (PACU), in the phase II unit, and 24 h after surgery. We compared patients who received neostigmine (n = 40) for reversal of residual neuromuscular blockade with those who did not (n = 60). More patients receiving rocuronium required reversal drugs than those receiving mivacurium (68% vs 10%). There were no differences in the incidence of nausea (18% vs 15%), vomiting (8% vs 12%), and the need for antiemetics (13% in both the groups) in the PACU between patients who received neostigmine and those who did not. In addition, the duration of PACU stay and the time to home-readiness were also similar between the groups. We conclude that, compared with rocuronium, the use of mivacurium decreases the need for reversal drugs. In addition, reversal of residual neuromuscular blockade with neostigmine does not increase the incidence of PONV or the need for antiemetic medications in patients undergoing ambulatory surgery. IMPLICATIONS: In this study, we showed that the incidence of postoperative nausea and vomiting and the need for antiemetics do not increase with the use of neostigmine and glycopyrrolate for reversal of residual muscle paralysis. PMID- 10475293 TI - The efficacy of a simulated intravascular test dose in sevoflurane-anesthetized children: a dose-response study. AB - A recent study demonstrated that changes in both heart rate (HR; positive if > or = 10bpm increase) and T-wave amplitude (positive if > or = 25% increase) reliably detect accidental intravascular injection when a full test dose containing epinephrine 0.5 microg/kg is injected intravascularly. We designed this study to prospectively determine whether a smaller dose of epinephrine would produce reliable HR and T-wave changes in sevoflurane-anesthetized children. We studied 80 ASA physical status I infants and children (6-72 mo) undergoing elective surgeries during 1.0 minimum alveolar anesthetic concentration sevoflurane and 67% nitrous oxide in oxygen. After the administration of i.v. atropine 0.01 mg/kg, the patients were randomly assigned to receive either i.v. saline (n = 20), an i.v. test dose (0.1 mL/kg) consisting of 1% lidocaine with 1:200,000 epinephrine (epinephrine 0.5 microg/kg group, n = 20), an i.v. test dose (0.05 mL/kg) (epinephrine 0.25 microg/kg group, n = 20), or an i.v. test dose (0.025 mL/kg) (epinephrine 0.125 microg/kg group, n = 20) via a peripheral vein to simulate the intravascular injection of the test dose. HR and systolic blood pressure were recorded every 20 and 30 s, respectively, and T-wave amplitude of lead II was continuously recorded for subsequent analysis. After the i.v. injection of the test dose, all children in the epinephrine 0.5 and 0.25 microg/kg groups developed positive responses based on the peak T-wave amplitude, whereas all children in the epinephrine 0.5 microg/kg group and 17 children (85%) in the epinephrine 0.25 microg/kg group elicited a positive response according to the peak HR criterion. No false-positive responses were observed with saline injections. Children in the epinephrine 0.125 microg/kg group showed clinically unacceptable efficacy based on either criterion. We conclude that the efficacies of detecting an intravascular injection of the test dose based on the hemodynamic and T-wave criteria are reduced with smaller doses of epinephrine and that HR and T-wave changes are still useful indicators in most patients if epinephrine 0.25 microg/kg is accidentally injected intravascularly. IMPLICATIONS: To determine whether an epidurally administered local anesthetic has been unintentionally injected into a blood vessel, a small dose of epinephrine is often added to a local anesthetic. We found that an increase in T-wave amplitude > or = 25% in lead II and a heart rate increase > or = 10 bpm are useful indicators for detecting the accidental intravascular injection of a small dose of epinephrine in sevoflurane-anesthetized children. PMID- 10475294 TI - Pressure-limited ventilation of infants with low-compliance lungs: the efficacy of an adult circle system versus two free-standing intensive care unit ventilator systems using an in vitro model. AB - We compared the efficacy of a Drager Narkomed GS (North American Drager, Telford, PA) equipped with an adult circle system with two free-standing infant ventilator systems (Servo 300; Siemens Medical Systems, Danvers, MA and Babylog 8000; North American Drager) to deliver minute ventilation (VE) using pressure-limited ventilation to a test lung set to low compliance. To simulate a wide variety of potential patterns of ventilation, VE was measured at peak inspiratory pressures (PIP) of 20, 30, 40, and 50 cm H2O and at respiratory rates (RR) of 20, 30, 40, and 50 breaths/min. Each measurement was made three times; the average was used for data analysis using the multiple regression technique. Delivered V(E) was positively correlated with both PIP (P = 0.001) and RR (P = 0.001). Only minimal differences in VE were observed between the circle and the two free-standing systems. At lower RR and PIP, the Babylog 8000 system delivered slightly higher VE than the circle system, whereas at higher RR and PIP, the Babylog 8000 delivered slightly lower VE than the circle system; these differences in VE were not statistically significant (P = 0.45). The Servo 300 delivered slightly higher VE than the circle system in all test conditions, but these differences were not statistically significant (P = 0.09). None of the differences in delivered VE between the Servo 300 and the circle system are of clinical importance. IMPLICATIONS: Our laboratory investigation suggests that pressure-limited ventilation delivered by a standard adult circle system compares favorably with that of freestanding infant ventilators used in pressure-limited mode. Changing from an adult circle system to a free-standing pressure-limited ventilator may not substantially improve ventilation of a low-compliance infant lung; the efficacy of such a practice should be investigated. PMID- 10475295 TI - Neonatal resuscitation with the laryngeal mask airway in normal and low birth weight infants. PMID- 10475296 TI - The imposed work of breathing is less with the laryngeal mask airway compared with endotracheal tubes. PMID- 10475297 TI - Levobupivacaine for ilioinguinal/iliohypogastric nerve block in children. PMID- 10475298 TI - Intraoperative breathing circuit obstruction caused by albuterol nebulization. PMID- 10475299 TI - Which clinical anesthesia outcomes are important to avoid? The perspective of patients. AB - Healthcare quality can be improved by eliciting patient preferences and customizing care to meet the needs of the patient. The goal of this study was to quantify patients' preferences for postoperative anesthesia outcomes. One hundred one patients in the preoperative clinic completed a written survey. Patients were asked to rank (order) 10 possible postoperative outcomes from their most undesirable to their least undesirable outcome. Each outcome was described in simple language. Patients were also asked to distribute $100 among the 10 outcomes, proportionally more money being allocated to the more undesirable outcomes. The dollar allocations were used to determine the relative value of each outcome. Rankings and relative value scores correlated closely (r2 = 0.69). Patients rated from most undesirable to least undesirable (in order): vomiting, gagging on the tracheal tube, incisional pain, nausea, recall without pain, residual weakness, shivering, sore throat, and somnolence (F-test < 0.01). IMPLICATIONS: Although there is variability in how patients rated postoperative outcomes, avoiding nausea/vomiting, incisional pain, and gagging on the endotracheal tube was a high priority for most patients. Whether clinicians can improve the quality of anesthesia by designing anesthesia regimens that most closely meet each individual patient's preferences for clinical outcomes deserves further study. PMID- 10475300 TI - Evolution of the French public's knowledge and attitudes regarding postoperative pain, cancer pain, and their treatments: two national surveys over a six-year period. AB - Pain management has become a notable feature of public health policy and mass media communication in France over the past few years. To assess the evolution of the knowledge and attitudes of the French population with respect to pain management and morphine use, telephone surveys using similar questionnaires were conducted in 1990 (n = 1001) and 1996 (n = 1006). The proportion of respondents who would take pain management adequacy into consideration when selecting a surgical facility increased from 52% to 81% (P < 0.001), as did the proportion who associated morphine with pain treatment (from 44% to 80%; P < 0.001) or who would not be afraid of becoming addicted to morphine after it had been prescribed for pain relief (from 26% to 69%; P < 0.001). However, the proportion of respondents who agreed that morphine can be prescribed to patients with pain increased only slightly. In 1996, 58% of the respondents believed that their knowledge had improved over the past 5 yr and associated this improvement first with television, followed by written press articles and by interaction with physicians. Increased awareness of pain management possibilities among the public may generate increased demand on health professionals to provide adequate and precise information addressing each patient's needs. IMPLICATIONS: The results of two representative surveys conducted over a 6-yr interval show significant improvements of knowledge and attitudes regarding pain and its management in the French general population. However, these results point to the need for additional specific information that should be provided through patient-physician interactions. PMID- 10475301 TI - Ketamine suppresses proinflammatory cytokine production in human whole blood in vitro. AB - The production of proinflammatory cytokines, such as tumor necrosis factor (TNF) a, interleukin (IL)-6, and IL-8, increases in patients with sepsis; marked production causes organ failure and septic shock. We previously reported that ketamine suppressed lipopolysaccharide (LPS)-induced TNF-alpha production in mice. However, there are no reports on the effect of ketamine on cytokine production in human whole blood. Therefore, in this study, we investigated the efficacy of ketamine on LPS-induced TNF-alpha, IL-6, and IL-8 production and recombinant human (rh) TNF-a-induced IL-6 and IL-8 production in human whole blood. After adding different doses of ketamine to whole blood, the blood was stimulated with LPS or rhTNF. After incubation, the plasma TNF-alpha activity and IL-6 and IL-8 concentrations were measured using the L929 cell cytotoxic assay or an enzyme-linked immunoassay. Ketamine significantly suppressed LPS-induced TNF alpha production at concentrations >20 microg/mL. At concentrations >100 microg/mL, ketamine also significantly suppressed both LPS-induced and rhTNF induced IL-6 and IL-8 production. In this study, we demonstrated that ketamine directly inhibits the production of proinflammatory cytokines such as TNF-alpha, IL-6, and IL-8 in human whole blood. IMPLICATIONS: We found that ketamine suppressed lipopolysaccharide-induced tumor necrosis factor alpha, interleukin (IL)-6, and IL-8 production and recombinant human tumor necrosis factor-induced IL-6 and IL-8 production in human whole blood. Ketamine directly suppresses proinflammatory cytokine production. PMID- 10475302 TI - Percutaneous tracheostomy and Murphy's law: an eye for trouble. PMID- 10475303 TI - Femoral nerve palsy: an unusual complication after femoral vein puncture in a patient with severe coagulopathy. PMID- 10475304 TI - Metabolic and hemodynamic changes during recovery and tracheal extubation in neurosurgical patients: immediate versus delayed recovery. AB - Delayed recovery has been advocated to limit the postoperative stress linked to awakening from anesthesia, but data on this subject are lacking. In this study, we measured oxygen consumption (V(O2)) and plasma catecholamine concentrations as markers of postoperative stress. We tested the hypothesis that delayed recovery and extubation would attenuate metabolic changes after intracranial surgery. Thirty patients were included in a prospective, open study and were randomized into two groups. In Group I, the patients were tracheally extubated as soon as possible after surgery. In Group II, the patients were sedated with propofol for 2 h after surgery. V(O2), catecholamine concentration, mean arterial pressure (MAP), and heart rate (HR) were measured during anesthesia, at extubation, and 30 min after extubation. V(O2) and noradrenaline on extubation and mean V(O2) during recovery were significantly higher in Group II than in Group I (V(O2) for Group I: preextubation 215 +/- 46 mL/min, recovery 198 +/- 38 mL/min; for Group II: preextubation 320 +/- 75 mL/min, recovery 268 +/- 49 mL/min; noradrenaline on extubation for Group I: 207 +/- 76 pg/mL, for Group II: 374 +/- 236 pg/ mL). Extubation induced a significant increase in MAP. MAP, HR, and adrenaline values were not statistically different between groups. In conclusion, delayed recovery after neurosurgery cannot be recommended as a mechanism of limiting the metabolic and hemodynamic consequences from emergence from general anesthesia. IMPLICATIONS: In this study, we tested the hypothesis that delayed recovery after neurosurgery would attenuate the consequences of recovery from general anesthesia. As markers of stress, oxygen consumption and noradrenaline blood levels were higher after delayed versus early recovery. Thus, delayed recovery cannot be recommended as a mechanism of limiting the metabolic and hemodynamic consequences from emergence after neurosurgery. PMID- 10475305 TI - Cerebral venous and tissue gases and arteriovenous shunting in the dog. AB - Cerebral venous blood gas values have been used to indicate brain tissue oxygenation. However, it is not clear how cerebral tissue and venous measures may vary under physiologic conditions caused by arteriovenous shunt. The purpose of this study was to measure brain tissue and local cerebral venous oxygen (PO2) and carbon dioxide (P(CO2)) partial pressure during changes in ventilation and to calculate shunt fraction. Eight dogs were anesthetized with isoflurane. After a craniotomy, a Neurotrend probe (Diametrics Inc., St. Paul, MN) that measures P(O2), P(CO2), pH, and temperature was inserted into brain tissue, and a small vein that drained the same tissue was catheterized. Arterial, cerebral venous, and brain tissue P(O2) and Pco2 were measured during random changes in ventilation to produce five different levels of inspired oxygen (room air, 40%, 60%, 80%, 95%) at each of three different end-tidal Pco2 (20 mm Hg, 40 mm Hg, 60 mm Hg). Arteriovenous shunt was calculated from oxygen and C(O2) content in artery, vein, and tissue, representing capillary. Tissue P(CO2) was 8 mm Hg greater than vein Pco2 during hypocapnia and this difference increased to 20 mm Hg during hypercapnia. Vein P(O2) was 8 mm Hg higher than tissue P(O2) during hypocapnia, and this difference increased to 40 mm Hg during hypercapnia. Shunt fraction increased from 10%-20% during hypocapnia to 50%-60% during hypercapnia. These results show that brain vein and tissue P(O2) and P(CO2) differ because of arteriovenous shunting and this difference is increased as end-tidal P(CO2) increases. IMPLICATIONS: We found, in dogs, that the gradient between brain venous and tissue P(O2) and PCO2 is increased with increased arterial P(CO2). The divergence between tissue and venous gases can be described by arterial to venous shunting. PMID- 10475306 TI - Nitrous oxide increases normocapnic cerebral blood flow velocity but does not affect the dynamic cerebrovascular response to step changes in end-tidal P(CO2) in humans. AB - We sought to clarify the effect of nitrous oxide (N2O) on the immediate responses of cerebral vasculature to sudden changes in arterial carbon dioxide tension in healthy humans. By use of a transcranial Doppler ultrasonography, blood flow velocity in the middle cerebral artery (V(MCA)) was measured during a step increase followed by a step decrease in end-tidal CO2 tension (PET(CO2)) between normo- and hypercapnia while subjects inspired gas mixtures containing 70%O2 + 30% N2 (control) and 70% O2 + 30% N2O (N2O) separately. During the control condition, both step increase and decrease in PET(CO2) produced rapid exponential changes in V(MCA). An increase in V(MCA) produced by the step increase in PET(CO2) was smaller (P < 0.001) and slower (P < 0.001) than a decrease in V(MCA) induced by the step decrease in PET(CO2). These general features of the dynamic cerebrovascular response were not affected by substitution of N2O for N2 in the inspired gases although N2O increased baseline V(MCA) by 15% (P < 0.001) compared with the control condition. We conclude that N2(O) in itself does not affect the dynamic cerebrovascular response to arterial CO2 changes, although it produces static mild cerebral vasodilation. IMPLICATIONS: This study suggests that nitrous oxide does not affect the dynamic cerebrovascular reactivity to acute arterial carbon dioxide (CO2) changes, i.e., exponential changes in cerebral blood flow in response to step changes in alveolar CO2 tension, although it does produce a mild increase in normocapnic cerebral blood flow velocity. PMID- 10475307 TI - The effects of propranolol on heterogeneity of rat cerebral small vein oxygen saturation. AB - beta-Adrenergic receptors are involved in altering cerebral metabolism and blood flow. This study was performed to determine whether propranolol would alter the microregional O2 balance in the brain. Rats were anesthetized with 1.4% isoflurane. Isotonic sodium chloride solution (control group), propranolol 2 mg/kg (low propranolol group) or propranolol 20 mg/kg (high propranolol group) was administered IV to the rats. Twenty minutes later, regional cerebral blood flow (rCBF) was measured using the 14C-iodoantipyrine autoradiographic technique. Small (diameter <70 microm) arterial and venous oxygen saturation (SaO2 and SvO2, respectively) was determined using microspectrophotometry in the alternate slices of the tissue sections used to measure rCBF. In both the low and high propranolol groups, average cortical rCBF was 35% lower than that in the control group. The average O2 consumption of the cortex of the propranolol groups was significantly lower than control (low propranolol: -41%, high propranolol: -49%). In all groups, SaO2 was almost identi-cal. The heterogeneity of the microregional SvO2 expressed as the coefficient of variation (CV = 100 x sD/mean) was significantly lower in the propranolol groups (low propranolol: 8.0+/-2.3, high propranolol: 7.3 +/- 2.9) than in the control group (13.4 +/- 3.5). The proportion of cortical veins with Svo2 <55% was significantly smaller in the low and high propranolol groups (4 of 60 and 3 of 60, respectively) than that in the control group (15 of 60). In the other brain regions, the data followed a similar pattern. Our data demonstrated that propranolol is effective in decreasing O2 consumption, improving microregional O2 balance, and reducing its heterogeneity in the brain. IMPLICATIONS: Our data suggest that the linkage of O2 supply and consumption is not tightly coupled under isoflurane anesthesia. beta-Adrenergic blockers may tighten this linkage and reduce the number of low O2-saturated microregions. PMID- 10475308 TI - The effects of N(G)-nitro-L-arginine-methyl ester on neurologic and histopathologic outcome after transient spinal cord ischemia in rabbits. AB - Little is known about the role of nitric oxide in the pathophysiology of spinal cord ischemia. We evaluated the effects of nitric oxide synthase (NOS) inhibition by N(G)-nitro-L-arginine-methyl ester (L-NAME) in rabbits whose abdominal aorta was occluded for 20 min (Experiment 1) or 25 min (Experiment 2). In Experiment 1, the L-NAME group (n = 6) received 3 mg/kg i.v. L-NAME, followed by an i.v. infusion of 3 mg x kg(-1). h(-1) until 6 h after reperfusion. Ischemia was induced 20 min after the start of L-NAME. The phenylephrine group (n = 6) received phenylephrine to maintain comparable blood pressure. The control group (n = 6) received saline. In Experiment 2, L-NAME (3 mg/kg i.v. L-NAME, followed by an i.v. infusion of 3 mg x kg(-1). h(-1) until 6 h after reperfusion) and phenylephrine groups (n = 6 each) were studied. Ischemia was induced 100 min after the start of L-NAME. Forty-eight hours after reperfusion, hindlimb motor function and histopathology of the spinal cord were examined. In Experiment 1, L NAME and phenylephrine both improved neurologic outcome, with higher intraischemic blood pressures than saline. In Experiment 2, L-NAME worsened the neurologic and histopathologic outcome compared with phenylephrine. Attenuation of damage by L-NAME in Experiment 1 may be attributable to an intraischemic blood pressure increase. The worse outcome with L-NAME in Experiment 2 suggests that NOS inhibition exacerbates ischemic spinal cord damage. IMPLICATIONS: Nonselective inhibition of nitric oxide synthase activity has aggravating effects on the neurologic and histopathologic outcome after transient spinal cord ischemia. PMID- 10475309 TI - A comparison of the placental transfer of ropivacaine versus bupivacaine. AB - This study compares the placental transfer of ropivacaine and bupivacaine using the dual perfused, single cotyledon human placental model. We studied the effects of maternal/fetal protein binding, maternal ropivacaine concentration, and fetal pH on ropivacaine transfer. At a clinically relevant maternal concentration (1 microg/mL), the calculated transfer ratios (local anesthetic percent transfer/antipyrine percent transfer) of ropivacaine (0.82 +/- 0.03) and bupivacaine (0.74 +/- 0.01) were comparable at the completion of the perfusion experiment (120 min). When the perfusates were modified to simulate actual in vivo plasma protein binding values, the maternal-to-fetal transfer of ropivacaine and bupivacaine decreased significantly (P < 0.05) as indicated by transfer ratios of 0.42% +/- 0.07% and 0.40% +/- 0.03%, respectively. No saturation of the transfer process was observed for either drug at the maternal concentrations investigated. The placental transfer of both local anesthetic agents increased significantly as the fetal pH decreased. This investigation shows that ropivacaine and bupivacaine cross the human placenta at a similar rate, despite their differences in lipophilicity and stereochemistry. Placental transfer of both compounds is highly influenced by maternal and fetal protein concentration and the fetal pH. IMPLICATIONS: The placental transfer of ropivacaine was shown to be similar to that of bupivacaine, and is thus highly influenced by the degree of maternal and fetal protein binding and fetal pH. PMID- 10475310 TI - Anesthetic management of the parturient with protein S deficiency and ischemic heart disease. PMID- 10475311 TI - The effectiveness of preemptive analgesia varies according to the type of surgery: a randomized, double-blind study. AB - The reliability of preemptive analgesia is controversial. Its effectiveness may vary among anatomical areas or surgical types. We evaluated preemptive analgesia by epidural morphine in six surgery types in a randomized, double-blind manner. Pain intensity was rated using a visual analog scale, a verbal report, and a measurement of postsurgical morphine consumption. Preemptive analgesia was effective in limb surgery and mastectomy, but ineffective for gastrectomy, hysterectomy, herniorrhaphy, and appendectomy. Relief of postsurgical pain in hemiorrhaphy was more rapid than that in the other surgery types. Preemptive analgesia was effective in limb surgery and mastectomy, but not in surgeries involving laparotomy, regardless of whether the surgery was major (gastrectomy and hysterectomy) or minor (herniorrhaphy and appendectomy). These results suggest that viscero-peritoneal nociception is involved in postsurgical pain. The abdominal viscera and peritoneum are innervated both heterosegmentally (in duplicate or triplicate by the vagus and/or phrenic nerves) and segmentally (by the spinal nerves). Therefore, supraspinal and/or cervical spinal neurons might be sensitized, despite the blockade of the segmental nerves with epidural morphine. The rapid retreat of the pain after hemiorrhaphy suggests that central sensitization remits soon after minor surgery, but that in appendicitis, it may be protracted by additional noxious stimuli, such as infection. IMPLICATIONS: Epidural preemptive analgesia was reliably effective in limb and breast surgeries but ineffective in abdominal surgery, suggesting involvement of the brainstem and cervical spinal cord via the vagus and phlenic nerves. PMID- 10475312 TI - Acute phase histopathological study of spinally administered midazolam in cats. AB - Midazolam may be a useful analgesic when administered intrathecally. However, neurotoxicity must be excluded. The purpose of this study was to investigate whether spinally administered midazolam induces acute-phase histopathological or inflammatory reactions of the spinal cord. A lumbar laminectomy was performed on 40 cats, and their spinal cords were exposed. Midazolam 10 mg (2 mL, n = 20 cats) or saline 2 mL (20 cats) was administered directly to the spinal cord. At 1, 2, 4, or 6 h after the administration, cats were killed, and the lumbar spinal cord was removed and fixed in 10% formalin. Histology was examined using light microscopy with hematoxylin and eosin staining. Both groups showed slight to moderate changes in the spinal cord, but no severe damage was observed. Inflammatory reactions were seen in only one cat in the saline group with slight neutrophil infiltration. These changes were not different between the midazolam group and the saline group. In conclusion, up to 6 h after direct exposure to midazolam, no acute histological damage or inflammatory reaction of the spinal cord was seen in cats. IMPLICATIONS: Spinally administered midazolam, even in large doses, does not cause acute neurotoxicity or inflammation of the spinal cord. PMID- 10475313 TI - The effect of spinal analgesia on visceral nociceptive neurons in caudal medulla of the rat. AB - A population of neurons resident in the caudal ventrolateral medulla are excited by noxious cutaneous and visceral stimuli from large portions of the body. These neurons act as monitors of ascending nociceptive information, and we hypothesized that they would be inhibited by spinally administered analgesics in a clinically relevant fashion. Rats were anesthetized with oxygen/ halothane. The caudal medulla was surgically exposed, and a catheter placed into the intrathecal space overlying the lower thoracic spinal cord via the surgical site. Single medullary neurons were characterized for responses to cutaneous and visceral (colorectal distension) stimuli. The effects of i.v. and intrathecally administered morphine and lidocaine were determined. The intrathecal infusion of morphine for 6 days before testing was also used as a pretreatment. Colorectal distension-evoked responses of medullary nociceptive neurons were inhibited in a dose-dependent, naloxonereversible fashion by intrathecal and i.v. morphine (50% effective dose values: 3.5 and 440 microg/kg, respectively). Intrathecal lidocaine abolished responses to colorectal distension and produced a spinal level at doses producing minimal effects when administered systemically. Prior treatment with an infusion of morphine produced tolerance to the effects of subsequent intrathecal morphine administration. These findings support the use of this preparation as a neurophysiologic model of spinal analgesia. IMPLICATIONS: Neurons in the brainstem, isolated electrophysiologically, were used as whole body monitors of pain-related activity in the rat. As a neurophysiologic model of nociception, this preparation may prove useful for the study of regionally administered analgesics and local anesthetics. PMID- 10475314 TI - Comparison of ropivacaine 0.2% and lidocaine 0.5% for intravenous regional anesthesia in volunteers. AB - A longer acting local anesthetic such as ropivacaine may offer advantages over lidocaine for IV regional anesthesia (IVRA). The objective of this investigation was to determine whether the use of ropivacaine improves the quality and duration of IVRA. In a randomized, double cross-over design, 10 volunteers received lidocaine 0.5% or ropivacaine 0.2% for IVRA of the upper extremity on two separate days with a standard double-cuff technique. Sensation to pinprick, response to tetanic stimuli, and tourniquet pain were assessed on a 0-10 verbal numeric score scale at 5-min intervals throughout the period of tourniquet inflation. Motor function was evaluated by grip strength. After release of the second (distal) cuff, pinprick sensation, motor strength, and systemic side effects were evaluated at 3, 10, and 30 min. No significant differences were observed for onset times of anesthesia and times to proximal (38 +/- 3 and 36 +/- 3 min) or distal (34 +/- 13 and 36 +/- 13 min) tourniquet release after the administration of ropivacaine and lidocaine, respectively. However, postdeflation hypoalgesia and motor blockade were prolonged with ropivacaine, and postdeflation light-headedness, tinnitus, and drowsiness were more prominent with lidocaine. We conclude that ropivacaine may be an alternative to lidocaine for IVRA. It may result in prolonged analgesia and fewer side effects after tourniquet release. IMPLICATIONS: In this study, volunteers received lidocaine 0.5% or ropivacaine 0.2% for IV regional anesthesia on two study days. Ropivacaine and lidocaine provided similar surgical conditions. However, after release of the distal tourniquet, prolonged sensory blockade and fewer central nervous system side effects were observed with ropivacaine. PMID- 10475315 TI - The relative potency of oral transmucosal fentanyl citrate compared with intravenous morphine in the treatment of moderate to severe postoperative pain. AB - Pharmacokinetic studies have shown that oral transmucosal absorption of fentanyl is relatively rapid compared with gastrointestinal absorption, and it results in increased bioavailability. We designed this study to establish the relative potency of oral transmucosal fentanyl citrate (OTFC) compared with i.v. morphine in 133 postoperative patients. The morning after surgery, patients randomly received one dose of either OTFC (200 or 800 microg) and a placebo i.v. injection or i.v. morphine (2 or 10 mg) and an oral transmucosal placebo unit. Pain intensity, pain relief, time to meaningful pain relief, and time to remedication were recorded. Median time to onset of relief was approximately 5 min for all groups. Over the first hour, little difference among treatment groups was seen for pain intensity and pain relief. By 2 h after study drug administration, 800 microg of OTFC and 10 mg of i.v. morphine generally produced similar analgesia, which was better than the smaller doses. Duration of analgesia with the larger doses (800 microg of OTFC and 10 mg of morphine) was similar and longer that produced by the smaller doses. The larger doses of OTFC and morphine produced better and more sustained analgesia than 200 microg of OTFC or 2 mg of morphine. IMPLICATIONS: The relative potency of oral transmucosal fentanyl citrate (OTFC) to i.v. morphine was 8-14:1. In this postoperative setting, OTFC produced rapid pain relief similar to that produced by i.v. morphine. The larger doses of OTFC (800 microg) and morphine (10 mg) produced better and more sustained analgesia than 200 microg of OTFC or 2 mg of morphine. PMID- 10475316 TI - Peribulbar anesthesia with either 0.75% ropivacaine or a 2% lidocaine and 0.5% bupivacaine mixture for vitreoretinal surgery: a double-blinded study. AB - No study has evaluated the efficacy of ropivacaine in peribulbar block for ophthalmic surgery. The purpose of this prospective, randomized, double-blinded study was to compare ropivacaine and a lidocaine-bupivacaine mixture in peribulbar anesthesia. Sixty ASA physical status I or II patients scheduled for elective vitreoretinal surgery were randomized to receive a peribulbar block with 8 mL of either 0.75% ropivacaine (ropivacaine group, n = 30) or a 1:1 mixture of 2% plain lidocaine and 0.5% plain bupivacaine (lido-bupivacaine group, n = 30). Time required for onset of surgical anesthesia, quality of postoperative analgesia, incidence of side effects, and analgesic consumption were recorded. Surgical block was achieved after 8 +/- 5 min in the lido-bupivacaine group and after 10 +/- 5 min in the ropivacaine group (P = 0.23). A 3-mL supplemental injection 15 min after block placement was required in 6 patients in the lido bupivacaine group (20%) and in 10 patients in the ropivacaine group (33%) due to inadequate motor block (P = 0.38). On Postoperative Day 1, 26 patients in the ropivacaine group (87%) reported no pain at the verbal rating score, compared with 18 patients in the lido-bupivacaine group (60%) (P = 0.005). We conclude that 0.75% ropivacaine may be a suitable choice when performing peribulbar anesthesia for vitreoretinal surgery. IMPLICATIONS: Quick onset of block with prolonged postoperative analgesia is an important goal in regional anesthesia for ophthalmic surgery. Evaluating clinical properties of 0.75% ropivacaine and a 1:1 mixture of 2% lidocaine and 0.5% bupivacaine for peribulbar anesthesia, we demonstrated that ropivacaine has an onset similar to that of the lidocaine bupivacaine mixture and provides a better quality of postoperative analgesia. PMID- 10475317 TI - Fentanyl pretreatment does not impair the reliability of an epinephrine containing test dose during propofol-nitrous oxide anesthesia. AB - We designed this study to determine the hemodynamic responses to and the efficacy of a simulated IV test dose during propofol anesthesia based on the conventional heart rate (HR; positive if increase > or =20 bpm), the modified HR (positive if increase > or =10 bpm), and the systolic blood pressure (SBP; positive if increase > or =15 mm Hg) criteria. Eighty healthy patients were randomized to receive 2 mg/kgpropofol or propofol plus fentanyl (100 microg) at the induction of anesthesia (n = 40 each). After endotracheal intubation, anesthesia in both groups was maintained with propofol 8 mg x kg(-1) x h(-1) and 67% nitrous oxide in oxygen. Each group of patients was further divided into a test dose group receiving 1.5% lidocaine 3 mL plus epinephrine 15 microg (1:200,000) or a saline group (n = 20 each) receiving 3 mL of isotonic sodium chloride solution i.v. HR and SBP were monitored for 4 min after the i.v. injection of the study drug. The i.v. injection of the test dose produced a HR increase > or =20 bpm in 20 and 17 patients in the propofol and propofol-fentanyl groups, respectively, whereas all patients receiving the test dose and none receiving saline had HR increases > or =10 bpm. Therefore, in the propofol-fentanyl group, sensitivity, specificity, positive predictive value, and negative predictive value were 85%, 100%, 100%, and 87% according to the conventional HR criterion, and all were 100% according to the modified HR criterion. In the propofol group, 100% efficacy was obtained based on both HR criteria. However, all patients receiving the test dose and none receiving saline developed a SBP increase > or =15 mm Hg, resulting in 100% efficacy based on the conventional SBP criterion in both groups. Our results indicate that both the modified HR criterion and the SBP criterion are clinically applicable during propofol anesthesia with or without supplemental fentanyl. IMPLICATIONS: To determine whether an epidural catheter is in a blood vessel, an epidural test dose containing 15 microg of epinephrine is used. We found that, during propofol anesthesia with or without fentanyl, a heart rate increase > or =10 bpm and a systolic blood pressure increase > or =15 mm Hg are reliable indicators for detecting accidental intravascular injection. PMID- 10475318 TI - The preoperative administration of intravenous dextromethorphan reduces postoperative morphine consumption. AB - We evaluated the effect of dextromethorphan on postoperative pain management. Sixty ASA physical status I-III female patients undergoing major abdominal surgery underwent standardized general anesthesia. Thirty patients received an i.v. infusion of dextromethorphan 5 mg/kg before anesthetic induction (Pre group), whereas the remaining 30 patients received the same volume of isotonic sodium chloride solution, followed by a postoperative i.v. infusion of dextromethorphan 5 mg/kg (Post group). Patients in the Pre group received the same volume of isotonic sodium chloride solution postoperatively. All patients were then treated with patient-controlled i.v. analgesia, which administered a 0.6-mg bolus of morphine on demand (maximal 4 h dose 20 mg). The mean visual analog pain score during cough or movement and at rest were similar in the two groups in the first 3 days postoperatively. However, Post group patients consumed more morphine than Pre group patients during the first 2 days (P < 0.01). The sedation scores, patient satisfaction, and the incidence of morphine-related side effects were similar between the two groups. We conclude that the preoperative administration of dextromethorphan 5 mg/kg reduces postoperative morphine consumption compared with postoperative administration. IMPLICATIONS: In this double-blinded study, we found that the preoperative administration of i.v. dextromethorphan 5 mg/kg, compared with postoperative administration, reduces postoperative morphine consumption, which may provide clinical evidence of preemptive or preventive analgesic effects of dextromethorphan. PMID- 10475319 TI - Lack of rapid development of opioid tolerance during alfentanil and remifentanil infusions for postoperative pain. AB - Studies in animals and volunteers have suggested the development of acute tolerance to opioid analgesics. In this article, we present data from patients who regulated their own target-controlled infusions of alfentanil and remifentanil to provide analgesia in the immediate postoperative period. Fifty one patients received alfentanil for 24 h after cardiac surgery, and 30 patients received remifentanil for 6 h after orthopedic surgery. Satisfactory analgesia, defined as a rating of < or =3 on an 11-point visual analog scale, was obtained by patients after each type of surgery. The target concentrations of the opioids required to produce postoperative analgesia and the cumulative opioid doses administered over the course of the clinical observation suggest there was no tolerance to the analgesic effects of the opioids. The requirements for both analgesic drugs in individual patients had a large variation (>200%). We conclude that our results may indicate an absence of tolerance to opioids in postoperative analgesia. Nonetheless, our data show that the postoperative requirement for these rapidly acting drugs is qualitatively similar to that for other opioids in that dosage escalation does not occur. IMPLICATIONS: The development of acute tolerance to opioid analgesics has been suggested based on experimental studies in animals and volunteers. Our report from patients who self-controlled their analgesic requirements by using target-controlled infusions of alfentanil and remifentanil for postoperative analgesia provides no evidence of tolerance to opioids. PMID- 10475320 TI - Potentiation of narcosis after intravenous lidocaine in a patient given spinal opioids. PMID- 10475321 TI - Mexiletine as an adjuvant analgesic for the management of neuropathic cancer pain. PMID- 10475322 TI - Postoperative obstructive apnea. AB - We studied electromyography (EMG) of the geniohyoid muscle (Gh) and diaphragm (Di) in 12 postoperative, premedicated (flunitrazepam 2 mg PO), asymptomatic patients who snored after recovering from general anesthesia, the induction of which was partly achieved by i.v. midazolam. After extubation of the trachea, integrated EMG activity of Gh (E-Gh(MTA)) and Di (E-Di(MTA)) were measured. For Gh, tonic and phasic activity were distinguished. Patients were studied during obstructive apnea, at the end of apnea, while breathing through an artificial Guedel airway, and during quiet breathing 5 min after flumazemil. All patients experienced episodes of postoperative upper airway obstruction and nine became apneic. Flumazenil restored consciousness and predominant tonic E-GhMTA associated with upper airway patency in all patients. Reduced tonic E-GhMTA characterized postoperative obstructive apnea. Resolution of apnea required a burst of both tonic and phasic E-GhMTA associated with intense E-Di(MTA). Breathing through the Guedal airway resulted in patent airway in 8 of 10 patients and was associated with low tonic and phasic E-GhMTA and reduced E-Di(MTA). In this study, we demonstrated that the tonic pharyngeal muscular support modulates airway patency in the postoperative period. Because it is reversed by flumazemil, benzodiazepines are certainly the main cause of airway obstruction in these patients. IMPLICATIONS: Upper airway obstruction during recovery from general anesthesia induced by i.v. midazolam is associated with low tonic pharyngeal muscular support, which modulates upper airway patency in the postoperative period. PMID- 10475323 TI - The elimination of sodium and potassium hydroxides from desiccated soda lime diminishes degradation of desflurane to carbon monoxide and sevoflurane to compound A but does not compromise carbon dioxide absorption. AB - Normal (hydrated) soda lime absorbent (approximately 95% calcium hydroxide [Ca(OH)2], the remaining 5% consisting of a mixture of sodium hydroxide [NaOH] and potassium hydroxide [KOH]) degrades sevoflurane to the nephrotoxin Compound A, and desiccated soda lime degrades desflurane, enflurane, and isoflurane to carbon monoxide (CO). We examined whether the bases in soda lime differed in their capacities to contribute to the production of these toxic substances by degradation of the inhaled anesthetics. Our results indicate that NaOH and KOH are the primary determinants of degradation of desflurane to CO and modestly augment production of Compound A from sevoflurane. Elimination of these bases decreases CO production 10-fold and decreases average inspired Compound A by up to 41%. These salutary effects can be achieved with only slight decreases in the capacity of the remaining Ca(OH)2 to absorb carbon dioxide. IMPLICATIONS: The soda lime bases used to absorb carbon dioxide from anesthetic circuits can degrade inhaled anesthetics to compounds such as carbon monoxide and the nephrotoxin, Compound A. Elimination of the bases sodium hydroxide and potassium hydroxide decreases production of these noxious compounds without materially decreasing the capacity of the remaining base, Ca(OH)2, to absorb carbon dioxide. PMID- 10475324 TI - A demonstration of the concentration and second gas effects in humans anesthetized with nitrous oxide and desflurane. AB - In the present study, we explored both the existence of and the basis for the concentration and second gas effects. Groups of six normocapnic patients were given one of three gas mixtures via a nonrebreathing system: 65% nitrous oxide (N2O) plus 4% desflurane; 5% N2O plus 4% desflurane; or 65% N2O plus 0.5% desflurane plus 2% xenon (Xe). End-tidal carbon dioxide (CO2) was held constant by adjustments in ventilation. Confirming the existence of the concentration effect, the end-tidal (F(A)) concentration of N2O increased toward the inspired (F(I)) concentration more rapidly (i.e., F(A)/F(I) increased more rapidly) when the inspired concentration was 65% than when it was 5%. The F(A)/F(I) for desflurane also increased more rapidly when desflurane was given with 65% rather than 5% N2O, confirming the existence of the second gas effect. The small uptake of the second gas (desflurane) did not influence its own F(A)/F(I) or that of N2O; that is, the administration of 4%, rather than 0.5%, desflurane did not increase the rate of rise of F(A)/F(I) of either N2O or desflurane. One of the bases of the concentration and second gas effects, a concentrating of residual gases, was confirmed: administration of Xe with 65% N2O produced an F(A)/F(I) for Xe that exceeded 1.0. Patient sex did not seem to influence the rate of rise of F(A)/F(I) of either N2O or desflurane. Finally, we unexpectedly found that, despite an equal solubility in blood, the rise in F(A)/F(I) for N2O exceeded that for desflurane, perhaps because of differences in tissue solubilities and intertissue diffusion. IMPLICATIONS: As predicted by the concentration and second gas effects, increasing the inspired concentration of nitrous oxide accelerated its rate of rise and the rate of rise of concurrently administered desflurane in humans. PMID- 10475325 TI - The effect of olprinone compared with milrinone on diaphragmatic muscle function in dogs. AB - We compared the effect of olprinone with milrinone on the contractility of fatigued diaphragms in dogs. Animals were divided into four groups of 10 each. In each group, diaphragmatic fatigue was induced by intermittent supramaximal bilateral electrophrenic stimulation at a frequency of 20 Hz applied for 30 min. After producing fatigue, Group I received only maintenance fluids; Group II was given a bolus injection (50 microg/kg) followed by continuous infusion (0.5 microg x kg(-1) x min(-1)) of milrinone; Group III was infused with olprinone (10 microg/kg initial dose plus 0.3 microg x kg(-1) x min(-1) maintenance dose); Group IV was infused with nicardipine (5 microg x kg(-1) x min(-1)) during olprinone administration. After the fatigue-producing period in each group, transdiaphragmatic pressure (Pdi) at low-frequency (20 Hz) stimulation decreased from the prefatigued values (P < 0.05), whereas there was no change in Pdi at high-frequency (100-Hz) stimulation. In Groups II and III, during study drug infusion, Pdi at both stimuli increased from fatigued values (P < 0.05). The increase in Pdi was larger in Group III than in Group II (P < 0.05). In Group IV, the augmentation of Pdi by olprinone was abolished in the fatigued diaphragm with an infusion of nicardipine. We conclude that olprinone is more effective than milrinone for the improvement of contractility in he fatigued diaphragm and that the potentiating mechanism of olprinone may be closely related to the transmembrane calcium movement. IMPLICATIONS: Diaphragmatic fatigue may contribute to the development of respiratory failure. Compared with milrinone, olprinone improves the contractility in fatigued diaphragm in dogs. PMID- 10475326 TI - The influence of renal function on the pharmacokinetics and pharmacodynamics and simulated time course of doxacurium. AB - Doxacurium's clearance (C1) is markedly decreased in patients with renal failure undergoing kidney transplantation. However, no studies have determined the influence of renal function (as assessed by creatinine clearance [CrCl]) on its pharmacokinetics in patients without renal failure. We studied 53 patients aged 19-59 yr. During N2O/isoflurane anesthesia, doxacurium was infused over 10 min, plasma was sampled for up to 6 h, and twitch tension was measured. A three compartment model was fit to plasma concentration data and an effect compartment model to twitch data. Mixed-effects modeling was used to determine the influence of covariates, including CrC1, on doxacurium's pharmacokinetic/pharmacodynamic parameters. Obesity decreased both doxacurium's Cl (1.1% per percent above ideal body weight [IBW]) and its neuromuscular junction sensitivity (0.4% per percent above IBW). Cl increased 0.6% per mL/min increase in CrCl. In addition, the rate constant for equilibration between plasma concentration and effect decreased 46% per 1% increase in isoflurane, central compartment volume decreased 86% per 1% increase in isoflurane concentration, and slow distributional Cl decreased 69% per mg/ 100 mL increase in serum albumin. Simulations showed that the latter two covariates influence the time course of bolus doxacurium administration minimally. Both obesity and renal dysfunction prolong doxacurium's recovery markedly. When dosing is based on IBW, effects of CrCl on neuromuscular recovery are smaller compared with dosing based on actual weight. Therefore, obese patients should be dosed based on IBW. No further dosage adjustment is necessary for patients with renal dysfunction; however, recovery will take longer in patients with moderate-to-severe renal dysfunction. IMPLICATIONS: We examined the factors influencing doxacurium's pharmacokinetic and pharmacodynamic characteristics. Both creatinine clearance and obesity significantly influence its time course. The effect of obesity is minimized if patients are dosed based on ideal body weight. PMID- 10475327 TI - Massive subcutaneous emphysema and sudden airway compromise after postoperative vomiting. PMID- 10475328 TI - Propofol is effective in chemotherapy-induced nausea and vomiting: a case report with quantitative analysis. PMID- 10475329 TI - Respiratory mechanics during laparoscopic cholecystectomy. PMID- 10475330 TI - A laryngeal mask airway "tip". PMID- 10475331 TI - Heart rate control and ischemia. PMID- 10475332 TI - Transesophageal echocardiographic diagnosis of aortic pseudoaneurysm after combined aortic valve replacement and coronary artery bypass graft surgery. PMID- 10475333 TI - Intraarticular "analgesics": are they safe? PMID- 10475334 TI - Combitube: a new proposal for a confusing nomenclature. PMID- 10475335 TI - Concept for easy fiberoptic intubation via a laryngeal airway mask. PMID- 10475336 TI - Salivary gland enlargement caused by chemical agents. PMID- 10475337 TI - Acute obstruction during manual ventilation caused by an end-tidal rubber cap in the reservoir bag. PMID- 10475340 TI - Female sexual dysfunction: incidence, pathophysiology, evaluation, and treatment options. PMID- 10475338 TI - The importance of dose-response in study design. PMID- 10475341 TI - Oral pharmacotherapy for erectile dysfunction: current perspectives. PMID- 10475342 TI - Urologic applications of gene therapy. PMID- 10475343 TI - Acellular collagen matrix as a possible "off the shelf" biomaterial for urethral repair. AB - OBJECTIVES: To determine whether a naturally derived collagen-based tissue substitute developed for urethral reconstruction would be suitable for urethral repair in an animal model. Several urethral conditions often require nongenital tissues for reconstruction, such as skin grafts or mucosal grafts from the bladder or buccal regions. However, the use of these tissues for urethroplasty may be associated with additional procedures for graft retrieval, prolonged hospitalization, and morbidity. METHODS: A ventral urethral defect was created in 10 male rabbits. The acellular collagen matrix, obtained and processed from porcine bladder submucosa, was trimmed and used to replace the urethral defect in an onlay fashion. Serial urethrography was performed pre- and postoperatively at 0.5, 1, 2, 3, and 6 months. The animals were sacrificed 0.5, 1, 2, 3, and 6 months after surgery. The retrieved implants were analyzed grossly, histologically, and with immunocytochemistry. RESULTS: All animals survived until being sacrificed without any noticeable voiding dysfunction. Serial urethrograms confirmed the maintenance of a wide urethral caliber without any signs of strictures. Gross examination at retrieval showed normally appearing tissue without any evidence of fibrosis. Histologically, the implanted matrices contained host cell infiltration and generous angiogenesis by 2 weeks after surgery. The presence of a confluent transitional cell layer was confirmed by immunocytochemical analyses using pancytokeratin antibodies. Anti-alpha actin antibodies demonstrated the migration of unorganized muscle fiber bundles 2 months after implantation and organized muscle bundles 6 months after implantation. CONCLUSIONS: The acellular collagen matrix appears to be a useful material for urethral repair in the rabbit. The matrix can be processed easily, has good characteristics for tissue handling and urethral function, and has the advantage of being an "off the shelf" material. PMID- 10475344 TI - Ablation of canine prostate using transurethral intraprostatic absolute ethanol injection. AB - OBJECTIVES: Despite extensive research involving numerous treatments for benign prostatic hyperplasia (BPH), the ideal modality has yet to be discovered. This study evaluated chemoablation of the prostate using transurethral intraprostatic absolute ethanol injection (AEI) in an in vivo canine model. METHODS: Eight mongrel dogs, 7 to 10 years old, underwent transurethral intraprostatic AEI with various ethanol volumes (10 to 26 mL/animal, mean 19.9). Injection was performed using a 20-gauge, passive deflection, hollow-core needle, introduced cystoscopically by way of a perineal urethrotomy. Oral antibiotics were administered perioperatively. Blood alcohol levels were determined. The canines were kept alive for 1 hour (n = 1), 7 days (n = 2), and 21 days (n = 5) after the treatment. The dogs were observed twice daily for a minimum of 30 minutes to determine continence. At least one spontaneous voiding was recorded at each observation. Before the dogs were sacrificed, the prostate and surrounding tissues were harvested, with gross and microscopic examination performed by a single pathologist. RESULTS: Seven and 21 days after AEI, the prostates demonstrated necrosis and cavity formation. Deep injection resulted in cavity formation in a subcapsular location. Superficial injection resulted in cavity formation that was confluent with the urethra and resulted in a widened urethral lumen. No complications directly related to AEI were seen, and systemic absorption of ethanol was minimal. CONCLUSIONS: AEI can effectively ablate prostatic tissue in canines with minimal systemic absorption. No disruption of the prostatic capsule or injury to the bladder urothelium and urethral sphincter was identified. Human studies of intraprostatic AEI for BPH adenomatous tissue chemoablation are ongoing at our institution. PMID- 10475345 TI - Immunolocalization of inducible and constitutive nitric oxide synthases in human bladder cancer. AB - OBJECTIVES: Nitric oxide (NO) is synthesized by the enzyme family of NO synthases (NOS) and plays an important role in tumor growth and angiogenesis. NO generation by inducible NOS (iNOS) also influences the cytotoxicity of macrophages and tumor induced immunosuppression. Before now, the expression of iNOS and constitutive NOS in bladder carcinoma tissue had not been determined. METHODS: Bladder carcinoma tissue specimens were procured from 18 patients (mean age 69.7 years) undergoing transurethral resection. In every patient, tumor biopsies were compared with biopsies of benign bladder regions. Histochemical NADPH-diaphorase staining and NOS immunohistochemistry were performed on all tissue specimens. RESULTS: Positive NADPH-diaphorase staining was detected in all sections from bladder carcinoma tissue. NOS immunohistochemistry showed a different pattern. The malignant epithelial cells were highly iNOS positive. Specimens of bladder mucosa outside of the malignant regions showed only a weak positive iNOS immunostaining. The endothelial cells of abundant precapillary vessels in the stroma of bladder tumors showed a highly positive endothelial NOS (eNOS) immunostaining compared with the stroma of nonmalignant bladder tissue. Neuronal NOS immunoreactivity was only found in nitrinergic fibers in the fibromuscular stroma. CONCLUSIONS: Bladder carcinoma tissue had a high iNOS content; benign tissue did not. NO generation from iNOS in the malignant epithelium and from eNOS in tumor stroma may play different roles in tumor angiogenesis and tumor-induced immunosuppression. PMID- 10475346 TI - Evaluation of a new once-daily formulation of oxbutynin for the treatment of urinary urge incontinence. Ditropan XL Study Group. AB - OBJECTIVES: To evaluate in a 1 6-center, single-treatment study once-daily controlled-release oxybutynin (Ditropan XL) for urinary urge incontinence. METHODS: Two hundred fifty-six participants with urge incontinence or mixed incontinence with a significant urge component were treated. After baseline measurements, participants converting from conventional oxybutynin started Ditropan XL at their previous oxybutynin dose; others started at 5 mg/day. Doses were adjusted until participants reached a maintenance dose that produced continence or the best balance between continence and side effects. This dose was continued for 12 weeks. Effectiveness was assessed by urinary diaries. RESULTS: Effectiveness was achieved across all doses studied (5 to 30 mg/day), with 70.8% of participants using maintenance doses of 5 to 15 mg/day. Mean urge incontinence episodes per week decreased from 18.8 at baseline to 3.9 in maintenance week 1, 2.7 in week 4, and 2.8 at the end of the study. For those participants who reported urge incontinence episodes at baseline but were free of urge incontinence at maintenance week 1, 31% remained free of urge incontinence at every subsequent assessment. Participants who converted from other medications showed symptomatic improvement after conversion. At some time during the study, 58.6% of participants reported dry mouth, with 23.0% of participants rating it moderate or severe. Only 1.6% of participants discontinued the medication because of dry mouth. CONCLUSIONS: Ditropan XL treatment reduced the number of incontinence episodes. Maximum benefit was demonstrated by maintenance week 4 and was sustained through 12 weeks of maintenance therapy. PMID- 10475347 TI - Quantifying comorbidity in a disease-specific cohort: adaptation of the total illness burden index to prostate cancer. AB - OBJECTIVES: Numerous studies have demonstrated the importance of comorbid illness when analyzing medical outcomes. The purpose of this study was to adapt a generic comorbidity index, the Total Illness Burden Index (TIBI), for use in men with prostate cancer, and to evaluate the usefulness of the new instrument in adjusting for the impact of comorbidity on functional outcomes in a prostate cancer cohort. METHODS: The TIBI uses patients' self-report of symptoms and diagnoses to determine not only the presence but also the severity of comorbidities in each of 16 body system domains. To create the TIBI-P (prostate cancer modification), some domains were added and others were modified according to clinical criteria. The TIBI-P was completed by 1638 men with prostate cancer followed up longitudinally in 29 urology practices in the United States. TIBI-P scores were calculated for each patient and analyzed with scores on the SF-36 quality-of-life questionnaire and with patient report of days confined to bed. RESULTS: After adjusting for age and income, lower SF-36 scale scores and increases in confinement to bed were associated with a greater burden of comorbid illness as measured by the TIBI-P, independent of the extent of prostate cancer. The TIBI-P explained 24% of the variance in the SF-36 physical functioning domain score. CONCLUSIONS: The TIBI-P is a powerful measure of the impact of comorbid illness on the quality of life and functioning among patients with prostate cancer. This index may prove valuable in research on clinical and economic outcomes of prostate cancer. PMID- 10475348 TI - Does the rate of extracorporeal shock wave delivery affect stone fragmentation? AB - OBJECTIVES: To evaluate the effect of the rate of shock wave delivery on stone fragmentation, because the optimal rate of shock wave administration has not yet been established. METHODS: Standard phantom, ball-shaped, ceramic stones were placed in a net-like basket with a hole size of 2.2 mm and immersed in a specially designed water bath coupled with the Econolith 2000 lithotripter. One hundred eighteen stones (mean diameter 9.5 mm) were used. Shock waves were delivered at rates of 30, 60, 90, 120, and 150 shocks/min and at intensities of 15, 20, and 22.5 kV (electrohydraulic). The number of shocks required for complete fragmentation, determined by all fragmented particles falling through the basket holes, was recorded. RESULTS: The most effective (fewer shocks needed for complete stone fragmentation) rate of shock wave delivery was 60 shocks/min. A statistically significant difference was demonstrated between the mean number of shocks required for complete stone fragmentation at the rate of 60 shocks/min and faster rates at all energy levels (P <0.01) but not between the rate of 60 shocks/min and the rate of 30 shocks/min at all energy levels. CONCLUSIONS: The rate of shock wave administration during extracorporeal shock wave lithotripsy seems to influence stone disintegration. We demonstrated that extracorporeal shock wave lithotripsy is most effective when waves are delivered at 60 shocks/min. PMID- 10475349 TI - Effective analgesia for extracorporeal shock wave lithotripsy: transcutaneous electrical nerve stimulation. AB - OBJECTIVES: Extracorporeal shock wave lithotripsy (ESWL) has revolutionized the treatment of urinary stone disease. However, the most appropriate analgesia offering pain-free treatment, minimal side effects, and adequate cost effectiveness remains to be established. This prospective study was performed to evaluate the efficacy of transcutaneous electrical nerve stimulation (TENS) during ESWL using third-generation lithotripters. METHODS: Two pairs of stimulator electrodes were placed paravertebrally at L1 and near the lithotripter shock tube before ESWL. Treatment was carried out as follows: (a) shock wave administration was begun (no current = sham TENS); (b) in the case of severe pain, TENS was begun; and (c) if patients experienced no pain relief, analgesic drugs were given intravenously. RESULTS: Of 149 patients, 92 (62%) did not need any analgesia (neither TENS nor medication). In 42 (72%) of the remaining 57 patients, a TENS-related, pain-relieving response was observed. ESWL-induced pain could be reduced by 39.2%. The degree of fragmentation assessed by two urologists was found to be 90% for patients receiving TENS compared with a retrospectively analyzed control group (94%, n = 100). CONCLUSIONS: Two different theories explaining TENS-related analgesia are known: segmental (spinal) and supraspinal (central) inhibition. Since we did not observe any analgesic effect in patients having both pairs of electrodes attached around the shock tube (n = 30), supraspinal inhibition obviously accounts for the abovementioned pain relief. We conclude that TENS is a noninvasive, cost-effective method to achieve side-effect free analgesia in ESWL using third-generation lithotripters. PMID- 10475350 TI - Is the laparoscopic approach justified in patients with xanthogranulomatous pyelonephritis? AB - OBJECTIVES: Xanthogranulomatous pyelonephritis (XGP) is an atypical form of chronic renal infection. The treatment of choice is open nephrectomy, which is challenging, given the extent of the disease and the not uncommon involvement of the renal hilum and contiguous structures. We compared our experience with laparoscopic nephrectomy for histologically confirmed XGP with the open approach. METHODS: Review of all nephrectomy specimens at Washington University School of Medicine from July 1990 to March 1998 disclosed 9 patients with a pathologic diagnosis of unilateral XGP, of whom 5 patients underwent laparoscopic nephrectomy and 4 underwent open nephrectomy. XGP was suspected preoperatively in 56% of the patients. RESULTS: For the laparoscopic group, the average operating room time was 360 minutes, average blood loss was 260 mL, and complications occurred in 60% of patients (1 conversion to open, 1 ileus, 1 pulmonary embolus). For the open group, the average operating room time was 154 minutes, average blood loss was 438 mL, and there were no complications. Both groups were similar regarding time to oral intake, analgesia requirement, hospital stay, and time to complete recovery. CONCLUSIONS: Our early experience demonstrates that the benefits of laparoscopic nephrectomy, at present, do not extend to patients with XGP. Conventional open surgery is quicker, associated with fewer complications, and results in a similar use of analgesics, hospital stay, and recovery time. PMID- 10475351 TI - Autologous blood use in percutaneous nephrolithotomy. AB - OBJECTIVES: Preoperative autologous blood (AUB) donation has decreased patient exposure to allogenic blood (ALB) products and associated infectious risk. The risk of contracting hepatitis C and human immunodeficiency virus is 1 in 103,000 and 1 in 678,000, respectively, after receiving 1 U ALB. Elective surgical procedures require surgeons to offer preoperative AUB donation in California. Unused AUB is discarded. We report our use of AUB obtained for percutaneous nephrolithotomy. METHODS: A retrospective study of 144 consecutive patients who underwent 193 percutaneous nephrolithotomies between January 1994 and April 1998 at one of four teaching hospitals at the University of California, San Francisco was performed. Preoperative AUB donation, transfusion rates, hemoglobin levels, blood use, and costs were analyzed. RESULTS: Ninety-six units of blood were collected from 63 patients (44%) and were available for 70 procedures (36%). The overall transfusion rate per procedure was 7%, with 13 patients receiving a total of 24 U, 7 AUB and 17 ALB. Eighty-nine units (92.7%) of AUB were discarded, and the transfusion rate in donors and nondonors was similar. There was no significant difference in preoperative hemoglobin or operative blood loss between donors and nondonors. The 13 transfused patients had a lower preoperative hemoglobin ( 11.5 versus 12.8 g/dL; P = 0.029) and higher operative blood loss as measured by hemoglobin level (3.2 versus 1.6 g/dL; P <0.001). Blood bank charges for ALB and AUB were $ 119/U and $244 to $498/U, respectively, depending on transportation and thawing charges. CONCLUSIONS: Routine preoperative blood donation adds substantial cost for minimal benefit, given the low infectious risk of ALB and the two- to fourfold higher cost of AUB. In our series, women had an increased incidence of blood transfusion compared with men. AUB donation may provide peace of mind but is rarely used and is discarded 93% of the time. PMID- 10475352 TI - Preliminary study on urinary cytokine levels in interstitial cystitis: does intravesical bacille Calmette-Guerin treat interstitial cystitis by altering the immune profile in the bladder? AB - OBJECTIVES: To obtain preliminary urinary cytokine data on subjects with active interstitial cystitis (IC), subjects with IC in remission after bacille Calmette Guerin (BCG), and control (non-IC) subjects. IC is a severe, debilitating bladder disease of unknown etiology and no cure. In controlled clinical trials, intravesical BCG has been shown to be an effective and durable treatment for IC. The durability of this treatment led us to speculate on the mechanism by which intravesical BCG may treat IC. Evidence exists that IC may be mediated by an abnormal immune profile within the bladder. Intravesical BCG is known to stimulate the immune system of the bladder. METHODS: Fresh voided urine was collected from 15 subjects with active IC, 9 subjects with IC who received intravesical BCG and had been in remission for an average of 2.6 years, and 11 non-IC subjects. The urine was immediately centrifuged, aliquoted, and frozen in liquid nitrogen. At the time of urine collection, a validated IC questionnaire was completed. The enzyme-linked immunosorbent assay technique was used to determine levels of urinary cytokines interleukin (IL)-2, IL-4, IL-6, IL-8, IL 10, IL-12, tumor necrosis factor (TNF), human granulocyte-macrophage colony stimulating factor (hGM-CSF), IL-1beta, and interferon-gamma (IFN-gamma). RESULTS: Cytokines IL-4, IL-10, IL-12, TNF, hGM-CSF, IL-1beta, and IFN-gamma were not detected. Significant elevations in symptom scores and IL-2, IL-6, and IL-8 were found in the urine of subjects with active IC compared with subjects with IC in remission and control subjects. The urinary cytokine levels and symptom scores were identical in the IC group who had received BCG and the control group. CONCLUSIONS: Elevations in symptom scores and urinary cytokine levels were seen in subjects with active IC, suggesting an abnormal immune profile in this disease. Subjects with IC in remission after receiving BCG had identical cytokine levels and symptom scores as non-IC control subjects. Intravesical BCG may be effective in treating IC by correcting an aberrant immune imbalance in the bladder, leading to long-term symptomatic improvement. A prospective study is ongoing to further investigate the role of the immune system in IC. PMID- 10475353 TI - Grading pelvic prolapse and pelvic floor relaxation using dynamic magnetic resonance imaging. AB - OBJECTIVES: With significant vaginal prolapse, it is often difficult to differentiate among cystocele, enterocele, and high rectocele by physical examination alone. Our group has previously demonstrated the utility of magnetic resonance imaging (MRI) for evaluating pelvic prolapse. We describe a simple objective grading system for quantifying pelvic floor relaxation and prolapse. METHODS: One hundred sixty-four consecutive women presenting with pelvic pain (n = 39) or organ prolapse (n = 125) underwent dynamic MRI. The "H-line" (levator hiatus) measures the distance from the pubis to the posterior anal canal. The "M line" (muscular pelvic floor relaxation) measures the descent of the levator plate from the pubococcygeal line. The "O" classification (organ prolapse) characterizes the degree of visceral prolapse beyond the H-line. RESULTS: The image acquisition time was 2.5 minutes per study. Each study cost $540. In the pain group, the H-line averaged 5.2 +/- 1.1 cm versus 7.5 +/- 1.5 cm in the prolapse group (P <0.001). The M-line averaged 1.9 +/- 1.2 cm in the pain group versus 4.1 +/- 1.5 cm in the prolapse group (P <0.001). Incidental pelvic pathologic features were commonly noted, including uterine fibroids, ovarian cysts, hydroureter, urethral diverticula, and foreign body. CONCLUSIONS: The HMO classification provides a straightforward and reproducible method for staging and quantifying pelvic floor relaxation and visceral prolapse. Dynamic MRI requires no patient preparation and is ideal for the objective evaluation and follow-up of patients with pelvic prolapse and pelvic floor relaxation. MRI obviates the need for cystourethrography, pelvic ultrasound, or intravenous urography and has become the study of choice at our institution for evaluating the female pelvis. PMID- 10475354 TI - Intravesical therapy with pertussis toxin before radical cystectomy in patients with bladder cancer: a Phase I study. AB - OBJECTIVES: To ascertain the side effects of intravesical instillation of pertussis toxin (PTX) because it inhibits tumor cell motility in vitro and in vivo and seems to be a promising therapeutic approach against cancer. METHODS: We initiated a Phase I study and measured the effect of intravesical instillation of PTX before radical cystectomy. Study end points were PTX-related side effects. PTX was instilled at five dose levels, starting with 14 microg and continuing to 72 microg. RESULTS: Fifteen patients, with a median age of 64 years, were included in the study. Intravesical instillation of PTX was without local or systemic side effects (grade 0, according to National Cancer Institute toxicity criteria). CONCLUSIONS: Treatment with PTX was safe and well tolerated without any significant local or systemic toxicity in dosages up to 72 microg. Therefore, the influence of PTX on local tumor should be evaluated in a Phase II study. PMID- 10475355 TI - Predictability of urodynamic findings based on the Urogenital Distress Inventory 6 questionnaire. AB - OBJECTIVES: The Urogenital Distress Inventory (UDI-6) is a validated 6-item questionnaire that assesses lower urinary tract symptoms, including incontinence, in women. Similar indexes developed in men to evaluate symptoms of benign prostatic hyperplasia have failed to show a relationship with urodynamic (UD) parameters indicating bladder outlet obstruction (BOO). In this study, we sought to determine whether UDI-6 responses could predict information obtained during UD evaluations. METHODS: All women referred to our clinic with lower urinary tract complaints who completed a UDI-6 questionnaire and subsequently underwent UD evaluation were included (n = 128). UD findings used for analysis included Valsalva leak point pressure (VLPP), maximum flow rate (Qmax), and detrusor pressure at Qmax (PdetQmax). BOO was defined as Qmax of 1 5 mL/s or less and PdetQmax of greater than 20 cm H2O; detrusor overactivity (DO) was defined as any rise in detrusor pressure associated with urge during filling. RESULTS: The most common chief complaints were incontinence (mixed, 26.6%; stress, 20.3%; and urge, 13.3%), urgency/frequency (14.1%), and symptomatic prolapse (10.1%). There was a moderate correlation between a positive response to question 3 (stress urinary incontinence [SUI]) and leakage with strain or cough during UD evaluation (correlation coefficient = 0.51). In fact, most patients answering that SUI was moderately or greatly bothersome were found to have stress-induced leakage during the UD evaluation (82%), which differed significantly from those who reported no bother (Fisher's exact test, P = 0.0006). Severity of leakage assessed by VLPP, however, did not correlate with the severity assessed by any question. With regard to BOO in women, most patients who answered that incomplete emptying was their most bothersome symptom had BOO (61%), and most women with a different main complaint were unobstructed (73%, P <0.002). Finally, 30 of 36 women who answered that leakage related to urgency was moderately or greatly bothersome were found to have DO, which was significantly different than the incidence of DO in women who did not report this complaint (correlation coefficient = 0.38, P <0.001). CONCLUSIONS: Unlike similar indexes used to assess lower urinary tract symptoms in men, specific items from the UDI-6 may provide predictive information regarding UD findings in women, particularly with regard to SUI, BOO, and DO. However, if VLPP is considered vital to planning treatment, UD studies will still be required, since no question could estimate the severity of incontinence as determined by serial VLPP measurement. PMID- 10475356 TI - High-grade inflammation in prostate cancer as a prognostic factor for biochemical recurrence after radical prostatectomy. Pathologist Multi Center Study Group. AB - OBJECTIVES: To assess the prognostic value of prostatic stromal inflammation in surgically treated localized prostate carcinoma for biochemical recurrence-free survival. METHODS: Stromal prostatic inflammation grading was studied in 161 patients who underwent radical prostatectomy for prostate cancer without involvement of the lymph nodes and who did not receive preoperative or postoperative radiotherapy or hormonal therapy until recurrence occurred. Inflammation was graded as high-grade inflammation if confluence of inflammatory cell infiltrate and/or glandular epithelium disruption associated with interstitial inflammatory infiltrate were present and as low-grade inflammation otherwise. Each specimen was graded separately first in the stroma surrounding nonmalignant glands and second in the stroma surrounding malignant glands. Biochemical recurrence based on serum prostate-specific antigen (PSA) level was defined as two successive PSA measurements greater than 1 ng/mL. RESULTS: Malignant tissue was significantly less involved in high-grade inflammation than benign adjacent tissue (9.3% and 19.9%, respectively; P <0.01). In a univariate Kaplan-Meier analysis, the 5-year recurrence-free survival rate for patients with high-grade and low-grade classified prostates was 61.0% and 66.7% in benign tissue and 27.0% and 65.3% in malignant tissue, respectively, with a significant difference between grades only in malignant tissue (P <0.02). In a multivariate analysis controlling for Gleason grade, preoperative serum PSA, pathologic stage, and inflammation grade in malignant tissue, the latter factor remained significantly predictive of biochemical recurrence (P = 0.03). CONCLUSIONS: Patients with high-grade inflammation surrounding malignant glands in radical prostatectomy specimens had significantly more postoperative biochemical recurrence than patients with low-grade inflammation. PMID- 10475357 TI - Comparison of once and twice daily dosage forms of Pygeum africanum extract in patients with benign prostatic hyperplasia: a randomized, double-blind study, with long-term open label extension. AB - OBJECTIVES: To compare the efficacy and safety of Pygeum africanum extract, 50 mg twice daily and 100 mg once daily. METHODS: Patients with symptomatic benign prostatic hyperplasia (BPH) entered a 2-month randomized, parallel-group, double blind, comparative phase (group A, 50 mg twice daily; group B, 100 mg once daily), followed by a 10-month, open phase (100 mg once daily). Main efficacy assessment parameters included International Prostate Symptom Score (IPSS), quality of life (QOL), and maximum urinary flow rate (Qmax). RESULTS: Two hundred nine patients completed the comparative phase in compliance with the protocol; 174 were included in the open phase. Both treatments had similar efficacy. IPSS (baseline 17 in both groups) improved by 38% in group A and 35% in group B. QOL improved by 28% in both groups. Qmax increased by 1.63 mL/s (16%) in group A and 2.02 mL/s (19%) in group B. After 12 months, the IPSS fell from 16 (baseline) to 9 (-46%). Half of the patients had an IPSS below 8. Mean Qmax increased by 1.65 mUs (15%). The safety profile was similar between groups and study phases. CONCLUSIONS: P. africanum extract at 50 mg twice daily and 100 mg once daily proved equally effective and safe at 2 months. Further improvements in efficacy with a satisfactory safety profile were documented after 12 months. PMID- 10475358 TI - Role of microvessel density in predicting recurrence in pathologic Stage T3 prostatic adenocarcinoma. AB - OBJECTIVES: Extraprostatic extension of prostatic adenocarcinoma (pathologic Stage T3) increases the risk of recurrence after radical prostatectomy compared with organ-confined prostate cancer. Use of microvessel density in predicting cancer recurrence in Stage pT3 cancer is poorly understood. We evaluated known predictors of recurrence, including Gleason grade, preoperative serum prostate specific antigen (PSA), DNA ploidy, seminal vesicle involvement, and surgical margin status in comparison with optimized microvessel density (OMVD) and area weighted microvessel density (AWMVD) in patients with Stage pT3 prostate cancer. METHODS: Between 1987 and 1989, 290 previously untreated patients underwent radical prostatectomy and were found to have pathologic Stage T3 adenocarcinoma. No patient received adjuvant therapy. Embedded prostatectomy specimens from 211 patients with sufficient tissue for immunohistochemical staining with factor VIII related antigen were studied by computer-assisted digital image analysis for OMVD and AWMVD. The correlation of Gleason grade, preoperative PSA, DNA ploidy, seminal vesicle involvement, surgical margin positivity, OMVD, and AWMVD with clinical or biochemical failure was assessed using the Cox proportional hazards model. Biochemical failure was defined as a postoperative increase in PSA greater than 0.2 ng/mL, and clinical failure was defined as a positive biopsy or metastasis on bone scan. RESULTS: The mean follow-up +/- SD for all patients was 7.1 +/- 1.8 years, with 43 deaths (9 due to prostate cancer) and 124 cases of clinical and/or biochemical recurrence. The mean OMVD was 65.0 +/- 17.3, and the mean AWMVD was 8.2 +/- 5.3. OMVD and AWMVD were not predictors of cancer recurrence or significantly associated with DNA ploidy or preoperative PSA. AWMVD was associated with Gleason grade (P = 0.003). The estimated relative risk (adjusted for other cancer variables) of clinical and biochemical recurrence associated with a change in OMVD from the 25th percentile (53.5) to the 75th percentile (75.4) was 1.14 (95% confidence interval 0.92 to 1.42). The estimated relative risk (adjusted) of clinical and biochemical recurrence associated with a change in AWMVD from the 25th percentile (4.8) to the 75th percentile (10.4) was 1.17 (95% confidence interval 0.97 to 1.42). Gleason grade, preoperative PSA, DNA ploidy, and seminal vesicle involvement were predictors of clinical and/or biochemical recurrence in univariate and multivariate analyses. CONCLUSIONS: Microvessel density, assessed by OMVD and AWMVD, did not predict recurrence in patients with pathologic Stage T3 adenocarcinoma of the prostate (TNM Stage T3N0M0). DNA ploidy, Gleason grade, preoperative PSA, and seminal vesicle involvement remained the best predictors of clinical and/or biochemical recurrence in this group of patients. PMID- 10475359 TI - Sensitivity and specificity of sextant biopsies in the detection of prostate cancer: preliminary report. AB - OBJECTIVES: To determine the true-negative and false-negative rates of sextant prostate biopsies, the most common method of prostate cancer diagnosis. METHODS: Forty-three men scheduled for prostatectomy as part of a surgical procedure for bladder pathologic findings agreed to participate in this study. All patients had normal digital rectal examination findings. Immediately before prostatectomy all patients underwent sextant biopsies. The location, amount, and Gleason grade of any cancer identified on the biopsies were recorded. After surgery, the prostate was serially sectioned. The location, grade, and volume of any prostatic adenocarcinoma identified was recorded and compared with the results of the biopsy specimens. RESULTS: There were 33 patients without prostate cancer in either the biopsies or the prostatectomy specimen. No patients had cancer on the biopsies and no cancer in the prostatectomy specimen. In 6 patients, cancer was found in both the biopsies and the prostatectomy specimens; these cancers were 0.9, 2.1, 2.8, 3. 1, 4.2, and 6.5 cc in volume. In the remaining 4 patients, there was no cancer on the biopsies but the prostatectomy specimen revealed cancers of 0.05, 0.1, 0.3, and 2.5 cc. The overall sensitivity for sextant biopsies was 60.0%, with a specificity of 100%. When only cancers greater than 2 cc or cancers in the peripheral zone were considered, the sensitivity rose to 83.3% and 71.4%, respectively, with a minimal decrease in specificity (97.3% and 97.2%, respectively). In contrast, when transition zone cancers were evaluated, the sensitivity fell to 33.3%. CONCLUSIONS: Sextant biopsies are fairly sensitive for the detection of tumors greater than 2 cc and those in the peripheral zone; however, repeat biopsies should be strongly considered in patients with a high clinical suspicion for prostate cancer and negative initial sextant biopsies. PMID- 10475360 TI - Which patients with newly diagnosed prostate cancer need a computed tomography scan of the abdomen and pelvis? An analysis based on 588 patients. AB - OBJECTIVES: Although a computed tomography (CT) scan of the abdomen and pelvis is often recommended as part of the staging evaluation for newly diagnosed prostate cancer, most scans are negative for metastases. We hypothesized that biopsy Gleason score, serum prostate-specific antigen (PSA) levels, and clinical stage could predict for a positive CT scan and that a low-risk group of patients could be identified in whom CT might be omitted. METHODS: All patients who had both pathologic review of their prostate cancer biopsies and abdominopelvic CT scans at our institution between January 1990 and May 1996 were studied. Gleason score, PSA, and stage were evaluated by univariate (chi-square) and multivariate (logistic regression) analyses for their ability to predict for a positive CT. RESULTS: Of 588 patients, 41 (7%) had a positive CT scan. Multivariate analysis showed Gleason score, PSA, and clinical stage to be significant independent predictors of a positive CT scan, all P <0.001. The odds ratios for a positive CT scan were 6.17 (95% confidence interval [CI] = 1.58 to 24) for Gleason score 8 to 10 versus 2 to 6; 2.25 (CI = 1.24 to 4) for PSA greater than 50 versus 0 to 15 ng/mL; 2.08 (CI = 1.70 to 3.21 ) for Stage T2c-T4 versus T2b or lower. All 244 patients with Gleason score 2 to 7, PSA 1 5 ng/mL or less, and clinical Stage T2b or less had negative CT scans. Of the other 174 patients with a Gleason score of 2 to 7, 8 (5%) had a positive CT scan. Of the 1 26 patients with a Gleason score of 8 to 10, 28 (22%) had a positive CT scan. CONCLUSIONS: Gleason score, PSA, and clinical stage were independent predictors for a positive CT scan of the abdomen and pelvis in patients with newly diagnosed prostate cancer. In this cost conscious era, we can decrease expenditure by obviating the need for a CT scan in low-risk patients (clinical Stage T2b or less, Gleason score 2 to 7, and PSA 15 ng/mL or less). A CT scan should be considered in all other patients. PMID- 10475361 TI - Does androgen suppression enhance the efficacy of postoperative irradiation? A secondary analysis of RTOG 85-31. Radiation Therapy Oncology Group. AB - OBJECTIVES: To evaluate the effect of immediate androgen suppression in conjunction with standard external beam irradiation (RT) versus RT alone on a group of men after prostatectomy who had indications for adjuvant treatment. METHODS: A national prospective randomized trial (Radiation Therapy Oncology Group [RTOG] 85-31) comparing standard external beam RT plus immediate androgen suppression versus external beam RT alone with delayed hormonal treatment at relapse was initiated for patients with locally advanced adenocarcinoma of the prostate. One hundred thirty-nine of the patients in this trial had indications for adjuvant treatment after prostatectomy (eg, capsular penetration, seminal vesicle involvement). Seventy-one of the patients received RT with immediate androgen suppression (luteinizing hormone-releasing hormone [LHRH] agonist); 68 patients received RT alone with hormonal manipulation instituted only at the time of relapse. RESULTS: With a median follow-up of 5 years, the estimated progression-free survival rate (failure defined as prostate-specific antigen [PSA] greater than 0.5 ng/mL) was 65% for the men who received combination therapy and 42% for those treated by RT alone with hormones reserved for relapse (P = 0.002). Differences in the rates of freedom from biochemical relapse were observed when failure was defined as PSA of 1.0 to 3.9 ng/mL (71% versus 46%; P = 0.008) and PSA greater than 4.0 ng/mL (76% versus 55%; P = 0.05), respectively. No differences were observed between the groups with respect to the end points of local control, distant failure, and overall survival. The use of immediate androgen suppression (ie, LHRH agonists) and the absence of pathologic nodal involvement were independently associated with prolongation of freedom from biochemical relapse by multivariate analysis. CONCLUSIONS: Patients with prostate cancer and indications for postoperative RT should be considered for combined RT and hormonal manipulation. Because statistically significant advantages for this experimental approach could not be defined for all end points studied (in particular, overall survival), efforts should be made to enroll these patients in the recently activated RTOG trial (96-01) comparing RT plus placebo to the combination of RT plus Casodex in the postoperative setting. PMID- 10475362 TI - Sexual function and bother after radical prostatectomy or radiation for prostate cancer: multivariate quality-of-life analysis from CaPSURE. Cancer of the Prostate Strategic Urologic Research Endeavor. AB - OBJECTIVES: To measure the effect of treatment choice (pelvic irradiation [XRT] versus radical prostatectomy [RP] with or without nerve sparing) on sexual function and sexual bother during the first 2 years after treatment. METHODS: We studied sexual function and sexual bother in 438 men recently diagnosed with early-stage prostate cancer and treated with XRT or RP with or without nerve sparing. Outcomes were assessed with the University of California, Los Angeles Prostate Cancer Index, a validated health-related quality-of-life instrument that includes these two domains. To minimize the influence of other factors, we adjusted for age, comorbidity, general health, and previous treatment for erectile dysfunction. All subjects were drawn from CaPSURE, a national, longitudinal data base. RESULTS: Sexual function improved over time during the first year in all treatment groups; however, during the second year, sexual function began to decline in the XRT group. Older patients who received XRT showed substantial declines in sexual function throughout the 2 years, and older patients who underwent RP experienced a return of very low baseline sexual function. Sexual function was improved by the use of nerve-sparing procedures or erectile aids. Alterations in sexual bother were ameliorated by many factors, including age, general health perceptions, and sexual function. CONCLUSIONS: Patients undergoing XRT or RP with or without nerve sparing all showed comparable rates of improvement in sexual function during the first year after treatment for early-stage prostate cancer. However, in the second year after treatment, patients treated with XRT began to show declining sexual function; patients treated with RP did not. PMID- 10475363 TI - Comparative quality-of-life analysis after radical prostatectomy or external beam radiation for localized prostate cancer. AB - OBJECTIVES: To determine and compare quality-of-life (QOL) evaluations from patients who received external beam radiation therapy or radical prostatectomy for the treatment of localized prostate cancer, and to compare differences in QOL assessments for urinary and sexual function after radical prostatectomy as reported by patient and physician. METHODS: Two hundred three patients treated by radical prostatectomy and 257 patients treated by external beam irradiation, all beyond 12-month follow-up after therapy, responded to a QOL questionnaire. The difference in responses with regard to bladder, bowel, and sexual function, overall satisfaction with treatment, and choice of the same treatment were assessed. Satisfaction with and choice of the same treatment were also specifically assessed according to bowel and bladder function and current disease status. The medical records of patients treated by radical prostatectomy were reviewed by an independent data manager to record the physician's assessment of continence and sexual function for comparison with that patient's assessment as noted in the questionnaire. RESULTS: Problems with urinary continence were more frequent among patients treated by radical prostatectomy; problems with gastrointestinal function were more frequent after irradiation. Sexual dysfunction was similar in both groups, although surgical patients experienced a greater impact on sexual relationships. The physician estimates of urinary continence were more favorable than the patient-reported outcomes. However, the physician estimate of sexual function closely approximated that of the patient. Preservation of sexual function among patients who underwent nerve-sparing surgery was disappointingly low. Only for the response to the question dealing with difficulty in achieving an erection was there a statistically significant benefit for patients receiving nerve-sparing versus non-nerve-sparing procedures. Patient satisfaction with and choice of the same treatment varied according to function and current disease status. Patients who had incontinence or bowel dysfunction or had evidence of recurrent disease were statistically less likely to choose the same treatment again when compared with functional and disease-free counterparts. Because irradiated patients were on average 6 years older than surgical patients, responses were adjusted for age; adjustment for age did not alter results. CONCLUSIONS: QOL is determined by the treatment received, by the assessment source, and by the patient's function and disease status at the time of assessment. Prospective and longitudinal studies will more accurately quantify immediate and chronic alterations in QOL. Uniformity of evaluation through consolidation of QOL instruments will permit more accurate cross-series and cross treatment comparisons. PMID- 10475364 TI - PSA, PSA density, PSA density of transition zone, free/total PSA ratio, and PSA velocity for early detection of prostate cancer in men with serum PSA 2.5 to 4.0 ng/mL. AB - OBJECTIVES: To enhance the specificity of prostate cancer (PCa) detection and reduce unnecessary biopsies in men with prostate-specific antigen (PSA) levels of 2.5 to 4.0 ng/mL, we prospectively evaluated various PSA-based diagnostic parameters. METHODS: This study included 273 consecutive men with serum PSA of 2.5 to 4.0 ng/mL referred for early PCa detection or lower urinary tract symptoms. All men underwent prostate ultrasound and sextant biopsy with two additional transition zone (TZ) biopsies. If the first biopsies were negative, repeated biopsies were performed at 6 weeks. Total PSA, PSA density (PSAD), PSA density of the transition zone (PSA-TZ), free/total PSA ratio (f/t PSA), and PSA velocity (PSAV) were determined, and the sensitivity, specificity, and predictive values of these various parameters were calculated. RESULTS: Of 273 patients, 207 had histologically confirmed benign prostatic hyperplasia (BPH) and 66 had PCa. f/t PSA and PSA-TZ were the most powerful predictors of PCa, followed by PSA, PSAD, and PSAV. Areas under the receiver operating characteristic curves for f/t PSA and PSA-TZ were 74.9% and 70.1%, respectively. With a 95% sensitivity for PCa detection, an f/t PSA cutoff of 41% and a PSA-TZ cutoff of 0.095 would result in the lowest number of unnecessary biopsies (29.3% and 17.2% specificity for f/t PSA and PSA-TZ, respectively) compared with all other PSA-related parameters evaluated. CONCLUSIONS: Compared with standard total PSA assays, f/t PSA and PSA TZ significantly enhance the sensitivity and specificity of PCa detection in a referral patient population with a total PSA of 2.5 to 4.0 ng/mL. PMID- 10475365 TI - Plasma levels of vascular endothelial growth factor are increased in patients with metastatic prostate cancer. AB - OBJECTIVES: Vascular endothelial growth factor (VEGF) is a cytokine that plays an important role in tumor angiogenesis. VEGF is overexpressed in many human cancers, including prostate cancer, but circulating levels of VEGF in patients with prostate cancer have not been reported. In this study, we analyzed plasma concentrations of VEGF in a cohort of patients with prostate cancer and compared them with a normal population. METHODS: Twenty-six healthy, cancer-free individuals and 80 patients with prostate cancer (54 patients with localized prostate cancer and 26 patients with metastatic prostate cancer [bone or lymph node positive]) were analyzed in this study. Blood was drawn in the same fashion from all individuals and deposited in tubes containing ethylenediaminetetraacetic acid as anticoagulant. Plasma was extracted and VEGF concentrations were determined using a quantitative sandwich enzyme immunoassay technique. RESULTS: Median plasma VEGF was 28.5 pg/mL (interquartile range 19.3 to 57.0) in patients with metastases; 7.0 pg/mL (interquartile range 0 to 26.5) in patients with localized disease, and 0 pg/mL (interquartile range 0 to 24) in controls. These differences were statistically significant (P <0.001). When compared group by group, the metastatic group had significantly higher plasma VEGF than the localized disease group and the control group (P = 0.003 and P <0.001, respectively). There was a tendency for plasma VEGF to be higher in the localized disease group than in the control group, a trend that almost reached statistical significance (P = 0.038). Using a cutoff of 18 pg/mL, the sensitivity and specificity of the test in differentiating between patients with and without metastatic disease was 81% and 71%, respectively. The odds of metastatic disease were almost 10 times greater for patients with VEGF values greater than 18 pg/mL than for those with values less than 18 pg/mL. There was no correlation between age and plasma VEGF values or between plasma VEGF and serum prostate-specific antigen (PSA). However, patients with serum PSA greater than 20 ng/mL had significantly higher plasma VEGF values than patients with serum PSA less than 20 ng/mL (P <0.001). No direct relation was found between Gleason sum and plasma VEGF, although VEGF levels were higher in patients with Gleason sums of 8 to 10 than in patients with lower Gleason sums. CONCLUSIONS: Our study indicates that patients with metastatic prostate cancer have higher plasma VEGF levels than patients with localized disease or healthy controls. A larger prospective study is needed to confirm the predictive utility of VEGF. PMID- 10475366 TI - Significance of small foci of Gleason score 7 or greater prostate cancer on needle biopsy. AB - OBJECTIVES: With increased screening for prostate cancer, we have noted a greater number of patients with small foci of Gleason score 7 or greater prostate cancer on needle biopsy. The significance of these findings is unknown. METHODS: We studied 57 men with small foci of Gleason score 7 or greater on needle biopsy. Tumor length was less than 1.5 mm in all but 2 cases. In those 2 cases, there were two minute (less than 0.5 mm) foci of cancer separated by 1.8 mm. The length of cancer ranged from 0.2 to 1.8 mm (mean 0.63 mm). In all cases, only one core was involved. RESULTS: Thirty-three men underwent radical prostatectomy (RP), 14 received radiation, 8 underwent surveillance, and 2 received hormonal therapy. Men who underwent RP were younger (62 years) than those who had radiotherapy (69.1 years), who were younger than those who underwent surveillance (74.5 years). The mean prostate-specific antigen (PSA) for men undergoing RP was 8.0 ng/mL (range 1.4 to 22). Preoperative serum PSA values did not predict organ confined status. Needle biopsy grades were as follows: 3 + 4 = 7 (n = 30); 4 + 3 = 7 (n = 17); 4 + 4 = 8 (n = 7); 5 + 4 = 9 (n = 1); and 5 + 5 = 10 (n = 2). We were able to review slides in 27 of the RP specimens, of which 24 were well sampled. Of these 24 cases, 33% had positive margins and 33% were not organ confined; the median tumor volume was 0.5 cc (mean 1.04). No difference in RP tumor volume was found between tumors with needle biopsy Gleason primary grade 3 and those with 4 or greater. The percentage of Gleason pattern 4 on needle biopsy weakly correlated with the percentage of Gleason pattern 4 in the RP specimen (P = 0.04). However, the percentage of Gleason pattern 4 only in the RP specimen, but not in the biopsy, correlated with whether the tumor was organ confined. CONCLUSIONS: The likelihood of having organ-confined disease with small foci of Gleason score 7 or greater on needle biopsy appears to be equivalent to that calculated from the Partin Tables for greater amounts of Gleason score 6 cancer on needle biopsy. In men who are considering RP, small foci of Gleason score 7 or greater adenocarcinoma on needle biopsy should not necessarily be considered an adverse finding. PMID- 10475367 TI - Age as a prognostic factor for disease recurrence after radical prostatectomy. AB - OBJECTIVES: To assess whether age has an impact on biochemical recurrence after radical prostatectomy for localized adenocarcinoma of the prostate. METHODS: Four hundred eighty-nine consecutive patients who underwent radical retropubic prostatectomy and did not have metastases to the lymph nodes were retrospectively analyzed. Disease recurrence was defined as a serum prostate-specific antigen greater than 0.2 ng/mL and rising on at least two postoperative measurements. Biochemical progression was compared in patients 70 years old or younger and older than age 70. The Kaplan-Meier estimator and Cox's proportional hazards model were employed to investigate the impact of age on time to recurrence. Neoadjuvant androgen deprivation was treated as a stratification variable in the Cox models. RESULTS: The mean follow-up was 25.4 +/- 20.8 months. The Gleason score and extent of cancer in the pathologic analyses of the prostatectomy specimen was not significantly different between the two groups. Biochemical recurrence was detected in 12% of patients 70 years old or younger and in 25% of those older than 70 (P = 0.01). In multivariate analyses, after adjusting for all prognostic factors, younger age (70 years or younger) was independently associated with a longer time to recurrence (P <0.02). CONCLUSIONS: Our results suggest that age per se may be an independent prognostic factor for disease recurrence after radical prostatectomy. This implies that patients 70 years old or younger are more likely to benefit from surgery. This information may be useful when counseling patients with clinically localized carcinoma of the prostate. PMID- 10475368 TI - Variable response to intracavernous prostaglandin E1 testing for erectile dysfunction. AB - OBJECTIVES: Anxiety and apprehension may negatively influence the erectile response to the first intracavernous injection with vasoactive agents. This may result in too many false-positive diagnoses of vascular insufficiency if the first injection ever made in a patient is used for color Doppler duplex sonography (CDS) evaluation. METHODS: One hundred sixty-eight consecutive patients (aged 18 to 75 years) with erectile dysfunction underwent a standardized evaluation, including the intracavernous injection test (ICIT) stimulated with 10 microg prostaglandin E1. Responses were recorded on a four-point scale: no response = 0, tumescence = 1, rigidity sufficient for intercourse = 2, full erection = 3. ICIT was repeated after 10 days and combined with CDS. The clinically assessed response to ICIT was correlated with end-diastolic flow velocity. RESULTS: Of 168 patients, 114 (68%) responded equally to the first and second ICIT, but 45 (27%) had an improved response, from tumescence to full erection in the second test (P <0.0001); in 9 (5%), the response deteriorated. The overall mean response was 1.6 (95% confidence interval 1.5 to 1.7) and 1.9 (95% confidence interval 1.7 to 2.0) (P <0.0001) for the first and second test, respectively. Of 168 patients, 89 (53%) responded with erections sufficient for intercourse when tested the first time and 104 (62%) did so after the second injection. CONCLUSIONS: Erectile response to diagnostic intracavernous injection of prostaglandin E1 significantly improved in the second compared with the first test. Therefore, cautious interpretation of CDS is advised when patients are injected for the first time because too many false-positive tests may result. PMID- 10475369 TI - Short-term analysis of the effects of as needed use of sertraline at 5 PM for the treatment of premature ejaculation. AB - OBJECTIVES: Pharmacotherapy using selective serotonin reuptake inhibitors (SSRIs) for men with primary premature ejaculation is promising. In particular, the strategy of taking a pill "as needed" may offer an attractive option. To investigate the possibility of self-therapy for the treatment of premature ejaculation, we compared the efficacy of sertraline taken as needed with that of continuous medication. METHODS: Since 1996, we have treated 24 men with sertraline on an as needed basis for primary premature ejaculation. Sertraline was chosen from among the SSRIs because a large dose need not be divided and because peak plasma levels occur 4 to 8 hours after oral administration, making 5 PM a suitable time for administration (by which time a man may know whether sexual intercourse is likely to occur later that evening). Each patient was started on 50 mg daily for 2 weeks, and the dose was then adjusted to 50 or 100 mg on the day of intercourse only (PRN). RESULTS: After 6 weeks, 18 men were still taking medication, and 6 had dropped out. Among the 18, the mean ejaculation latency was 23 +/- 19 seconds before treatment, 5.9 +/- 4.2 minutes after 2 weeks of 50 mg daily, 5.1 +/- 3.8 minutes after 2 weeks of 50 or 100 mg PRN, and 4.5 +/- 2.7 minutes after 4 weeks of 50 or 100 mg PRN. Mean sexual satisfaction scores (5, extremely satisfied; 0, extremely unsatisfied) for men were 0.8 +/- 0.8 before treatment, 3.8 +/- 1.2 after 2 weeks of 50 mg daily, 3.4 +/- 1.0 after 2 weeks of 50 or 100 mg PRN, and 3.2 +/- 0.7 after 4 weeks of 50 or 100 mg PRN. For their partners, mean sexual satisfaction scores were 1.1 +/- 0.7 before treatment, 3.2 +/- 1.6 after 2 weeks of 50 mg daily, 3.1 +/- 1.4 after 2 weeks of 50 or 100 mg PRN, and 3.3 +/- 1.2 after 4 weeks of 50 or 100 mg PRN. Side effects were intermittent excessive delay of ejaculation in 1 patient, fatigue in 2, and numbness in 1. CONCLUSIONS: If our results are supported by additional long-term clinical studies, self-therapy with sertraline taken PRN at 5 PM for the treatment of premature ejaculation could be as attractive as self injection therapy for the treatment of erectile dysfunction. PMID- 10475370 TI - Retroperitoneal recurrences after retroperitoneal lymph node dissection for low stage nonseminomatous germ cell tumors. AB - OBJECTIVES: To evaluate the incidence and sites of retroperitoneal recurrence after modified retroperitoneal lymph node dissection (RPLND) for low-stage nonseminomatous germ cell tumors (NSGCTs). METHODS: A retrospective review of 88 patients who underwent RPLND for Stage I or II NSGCTs between 1971 and 1991 was performed to determine the incidence and site of any retroperitoneal recurrence. RESULTS: Six retroperitoneal recurrences (four isolated and two in conjunction with failures at other sites) were found with a minimum follow-up of 5 years. All 6 patients had left-sided primary tumors, with three recurrences near the left renal hilum and two near the right renal hilum. All recurrences were at or outside the boundaries of the surgical dissection. Only one retroperitoneal recurrence may have been prevented by performing a bilateral dissection. CONCLUSIONS: Although uncommon, local recurrences can occur after RPLND. The right and especially the left renal hilum appear to be at higher risk of failure, possibly because of incomplete dissection in these areas. A more complete dissection in these areas may further decrease the local recurrence rate. Overall, the data presented and the studies reviewed suggest that modified RPLND reliably removes metastatic tumor with a low failure rate. If failures do occur, they are usually outside of the template and would not necessarily be prevented with a complete bilateral infrahilar dissection. PMID- 10475372 TI - Performing flexible ureteroscopy through a cystoscopy introducer sheath. AB - Performing flexible ureteroscopy through a cystoscope sheath positioned with its tip near the orifice of the ureter prevents buckling of the ureteroscope and decreases the friction against the ureteroscope. This results in more precise translation of the surgeon's movements to the tip of the instrument and facilitates the procedure. PMID- 10475371 TI - Gastric pouch acid secretion in response to physiologic digestive function. AB - OBJECTIVES: To evaluate acid secretion and histologic features of the gastric segment used for bladder replacement in bladder cancer. METHODS: Nine patients were investigated a mean of 36 months after gastric pouch surgery with modified shamfeeding and feeding. We determined urinary acidity, urinary pH, serum gastrin, and serum pancreatic polypeptide values and tried to find a relationship between feeding and acid secretion in the gastric pouch. In 6 patients, biopsy and histopathologic examination were performed. RESULTS: In 8 patients, urinary acidity increased after feeding following the rise of gastrin; in 1 patient no increase of gastrin or acidity was observed. In 7 patients, urinary acidity did not change after modified shamfeeding, indicating vagal denervation after surgery. This group showed a minimal urinary pH of above 4 after feeding. On the other hand, in 2 patients an increased acid secretion was observed after modified shamfeeding, indicating the possible presence of residual vagal innervation. In this group, the acid secretion in the pouch was higher, reaching the minimal pH level earlier after feeding. Histopathologic examination showed no major structural changes of gastric mucosa. CONCLUSIONS: Most patients are vagally denervated after gastric pouch surgery, and the gastric segment continues its original gastrointestinal function by a hormonal pathway. Our data indicate, however, that in some patients, the gastric pouch keeps a residual vagal innervation. We therefore suggest that nerve fibers present in the blood supply of the gastric segment be interrupted to avoid the complications associated with increased acid secretion of the gastric pouch. PMID- 10475373 TI - Computed tomography three-dimensional reconstruction in the diagnosis of traumatic renal artery thrombosis. PMID- 10475374 TI - Expression of CD44 adhesion molecules in nonpapillary renal cell carcinoma and normal kidneys. AB - OBJECTIVES: To analyze the relationship between the alterations in the expression of the CD44 gene in nonpapillary renal cell carcinoma (RCC) and several clinicopathologic factors. METHODS: The expression of the CD44 gene in 10 human RCC cell lines, 60 nonpapillary RCC tumor samples, and 15 normal kidney samples was investigated by reverse-transcription polymerase chain reaction (RT-PCR) using a set of primers capable of amplifying all CD44 variant isoforms. The results were analyzed with respect to several clinicopathologic factors. RESULTS: Nine of the 10 human RCC cell lines predominantly expressed the standard CD44 isoform (CD44s); CD44v10 was the major isoform in the 10th RCC cell line. The 15 normal kidney samples revealed the identical CD44 gene expression pattern; that is, CD44s, CD44v8-10, and CD44v10 were detectable in normal kidneys, and among them, CD44s was expressed most dominantly. In the 60 nonpapillary RCC samples, CD44s, CD44v8-10, and CD44v10 were the major isoforms in 46 (77%), 11 (18%), and 3 (5%) cases, respectively. Furthermore, the incidence of the predominant expression of CD44v8-10 in high-stage RCC was significantly higher than that in low-stage RCC. CD44s was more frequently expressed as a major isoform in clear cell RCC than in other histologic types of RCC. CONCLUSIONS: The results of this study show that the alternative splicing pattern of CD44 gene in RCC is different in each histologic type of RCC and suggest that CD44v8-10 upregulation in the progression of nonpapillary RCC is important. PMID- 10475375 TI - Expression of vascular endothelial growth factor receptors in human prostate cancer. AB - OBJECTIVES: We recently reported the expression and cytokine regulation of vascular endothelial growth factor (VEGF) in human prostate cancer (PCa). VEGF exerts its angiogenic and pro-tumorigenic properties by way of two high affinity receptors, fms-like tyrosine kinase 1 (FLT-1) and fetal liver kinase 1 (FLK-1). We hypothesized that these receptors are expressed and control VEGF functions in the PCa microenvironment. Herein, we evaluate the expression of these receptors in ex vivo PCa tissue, benign prostatic hypertrophy (BPH) tissue, and cultured PCa cell lines. METHODS: Ex vivo PCa specimens were obtained from patients undergoing radical retropubic prostatectomy. Specimens were selected to contain both PCa and BPH tissue (n = 15). Immunohistochemical analysis using antihuman FLT-1 and FLK-1 was performed and specimens were analyzed to characterize the expression and distribution of both receptors. Immunocytochemical analysis for FLT-1 and FLK-1 was also performed on cultured PCa cell lines (DU-145 and LNCaP). RESULTS: PCa cells expressed the VEGF receptor FLT-1 in 100% of specimens evaluated. Expression of FLK-1 was variable and related to tumor grade; high grade tumors displayed little or no FLK-1 expression. Vascular endothelial cells (VECs) within areas of PCa consistently expressed both FLT-1 and FLK-1 receptors. FLT-1 and FLK-1 were both expressed in BPH tissue. FLT-1 was expressed in the glandular epithelial cells in BPH, but in most cases FLK-1 was localized specifically to the basal cell layer of hypertrophic glands. FLT-1, but not FLK 1, was expressed by the DU-145 and LNCaP cell lines. CONCLUSIONS: Although they are differentially expressed, both FLT-1 and FLK-1 are present in PCa and BPH. Expression of receptors on VECs of tumor vessels supports the well-established role of VEGF in paracrine stimulation of VECs in the tumor microenvironment. The expression of FLT-1 and FLK-1 on tumor cells themselves suggests a potential autocrine function for VEGF (such as regulating tumor cell proliferation). These findings imply that a novel dual role may exist for VEGF, such that it is involved in tumor cell activation (autocrine), in addition to paracrine actions whereby it regulates endothelial cell functions and subsequent neovascular development. PMID- 10475376 TI - In vitro evaluation of calphostin C as a novel agent for photodynamic therapy of bladder cancer. AB - OBJECTIVES: Calphostin C, a highly specific protein kinase C inhibitor, induces apoptosis in the presence of visible light. We report the photoactivatable cytotoxicity of calphostin C in a series of well-characterized human bladder cancer cell lines: RT4, UM-UC-3, and 5637. METHODS: The human bladder cancer cell lines RT4, UM-UC-3, and 5637 were chosen on the basis of their p53, pRb and 9p21 deletion status. Using standard tissue culture techniques, the cytotoxicity of 10 to 100 nM calphostin C in combination with increasing exposures of visible light was examined. Controls consisted of cells treated with calphostin C without visible light and cells exposed to visible light without calphostin C treatment. Cell viability was determined by MTT assay. The induction of apoptosis by activated calphostin C was determined by 4,6-diamidino-2-phenylindole (DAPI) staining/fluorescence microscopy of nuclei. RESULTS: In the absence of light, calphostin C did not demonstrate a cytotoxic effect on any of the cell lines tested. Increasing the duration of light exposure resulted in a concomitant decrease in cell viability. Significant cell death was seen with calphostin C concentrations as low as 10 nM. These studies also demonstrated that calphostin C induced apoptosis by a mechanism independent of p53 and pRb status and the presence or absence of 9p21 deletions. CONCLUSIONS: We demonstrated the ability of activated calphostin C to induce apoptosis in a light-dependent and concentration-dependent fashion in a bladder cancer model system. Activated calphostin C cytotoxicity is independent of tumor genetic background and the status of p53 and pRb. Further development of calphostin C as a photosensitizer for photodynamic therapy of superficial bladder cancer may be warranted. PMID- 10475377 TI - Sildenafil in postmenopausal women with sexual dysfunction. PMID- 10475378 TI - Impact of sildenafil citrate (Viagra) on the DSM-IV diagnosis of male erectile disorder due to psychological factors. PMID- 10475379 TI - Detection of prostatic specific membrane antigen messenger RNA using immunobead reverse transcriptase polymerase chain reaction. AB - The present study was performed to detect circulating prostatic carcinoma (PC) cells using a novel three-step immunobead reverse transcriptase (RT) polymerase chain reaction (PCR) assay for prostatic specific membrane antigen (PSMA) messenger RNA (mRNA). The sensitivity and specificity of this technique was assessed and the incidence of immunobead RT-PCR positivity correlated with progressive metastatic disease and serum prostatic specific antigen (PSA) levels. Fifty peripheral blood (PB) samples from 46 patients with PC were incubated with magnetic beads coated with Ber-EP4 antibody directed against the human epithelial antigen a membrane antigen widely expressed by epithelial cells. The epithelial cell-enriched magnetic fraction was then subjected to mRNA isolation using oligo deoxythymidine (dT) magnetic beads. Nested RT-PCR for PSMA was performed on the mRNA oligo-dT complex and the identity of the RT-PCR products was confirmed by Southern blotting. Twenty-one PB samples from 8 control subjects without PC were also evaluated. Three-step immunobead PSMA RT-PCR was able to detect one PC cell per 1 mL of PB. The positivity rate of the RT-PCR assay was significantly higher (11 of 25; 44%) in patients with metastatic tumor than in patients with non metastatic disease (1 of 21; 5%) (P = 0.003). In patients with metastatic PC, RT PCR positivity was much higher in patients with progressive disease (10 of 13; 77%) than in patients with responding or stable disease (1 of 12; 8%) (P = 0.001). There was a statistically significant correlation between immunobead PSMA PCR positivity and high levels of serum PSA (P = 0.005). All control subjects without PC tested negative for PSMA PCR. The three-step immunobead RT-PCR for PSMA can detect circulating PC cells with high specificity and sensitivity. Preliminary data show a strong correlation between immunobead PCR positivity, the presence of progressive metastatic disease, and high levels of serum PSA. PMID- 10475380 TI - Rapid detection of mutated E-cadherin in peritoneal lavage specimens from patients with diffuse-type gastric carcinoma. AB - Tumor cells in abdominal lavage specimens from patients with gastric carcinoma strongly predict subsequent peritoneal metastasis and poor prognosis. Reverse transcription (RT)-polymerase chain reaction (PCR) detection of wild-type E cadherin has been claimed to be superior to conventional cytology for the detection of patients who subsequently develop peritoneal metastases. The present study tested this hypothesis and determined whether or not the detection of mutated, tumor-specific E-cadherin messenger RNA in abdominal lavage specimens serve as a useful diagnostic tool. Preoperative lavage specimens from 52 patients with diffuse-type gastric carcinoma and from 5 patients with benign disease were analyzed by conventional cytology and by RT-PCR for amplification of E-cadherin. Tumor cells were detected by cytology in 8 (15.3%) of the 52 patients with gastric cancer. The E-cadherin was detected in all 57 samples by RT-PCR. Two of these had abnormal E-cadherin amplification products confirmed to be mutations by direct sequencing, which were identical in the primary tumors. These findings suggest that the detection of wild-type E-cadherin is not sufficiently tumor specific. Also, for diffuse gastric carcinomas with confirmed E-cadherin mutations, detection of mutant E-cadherin by RT-PCR is a potentially valuable method for tumor cell detection in lavage specimens. PMID- 10475381 TI - Analysis of K-ras, N-ras, H-ras, and p53 in lung neuroendocrine neoplasms. AB - This study screened 11 samples of typical carcinoid (TC), 4 samples of atypical carcinoid (AC), 1 sample of large cell neuroendocrine carcinoma (LCNEC), and four metastases for point mutations in exons 5 to 8 of the p53 gene, and exons 1 and 2 of the K-ras. H-ras, and N-ras genes using polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) and direct sequencing and by immunohistochemistry for p53. Exon 1 of K-ras was mutated in two samples of low grade AC and a metastasis from one of these tumors (GAT12 and AGT12, respectively). No mutations in N-ras or H-ras were found. Mutations in exons 5 and 8 of the p53 gene were identified in a high-grade AC and a LCNEC. Positive immunostaining for p53 was present in three samples, with only one genotypic mutation shown (LCNEC). In conclusion, point mutations of the p53 gene were infrequent in these pulmonary neuroendocrine tumors, did not correlate in all samples with immunostaining, and were associated with the higher-grade tumors. Second, the presence of K-ras mutations seems to be associated with the higher grade carcinomas. Third, N-ras and H-ras mutations were not found with these pulmonary neuroendocrine tumors. PMID- 10475382 TI - Combined polymerase chain reaction approach for clonality detection in lymphoid neoplasms. AB - The present study analyzes the efficiency of a combination of four immunoglobulin heavy chain (IgH) gene polymerase chain reaction (PCR) primer systems and a multiplex T-cell receptor gamma chain (TRG) gene PCR for detection of clonality in 409 samples (234 paraffin sections, 175 bone marrow aspirates) of different lymphomas. Using the four IgH PCR systems together, clonality was detected in all samples of B-cell chronic lymphocytic leukemias, hairy cell leukemias, common acute lymphoblastic leukemias, and Burkitt-like B-cell lymphomas. Clonality was detected in all bone marrow aspirates with lymphoplasmacytoid immunocytoma, mantle cell lymphoma, marginal zone B-cell lymphoma, and unclassifiable low-grade B-cell lymphomas. The combined IgH gene PCR approach allowed clonality detection in 78.2% of myelomas, 75% of Burkitt lymphomas, 74.4% of diffuse large B-cell lymphomas, 68.7% of follicular center lymphomas, 50% of posttransplant lymphomas, 28.6% of anaplastic large cell lymphomas, 29% of T-cell lymphomas, and 18.8% of Hodgkin diseases. The combination of the four IgH gene primer systems with the multiplex TRG gene PCR allowed detection of clonality in 84.2% of B-cell neoplasms, 92.1% of T-cell non-Hodgkin lymphomas, and 18.8% of Hodgkin diseases, which was much more efficient than single PCR protocols. PMID- 10475383 TI - Definition of a region of loss of heterozygosity at chromosome 11q in cervical carcinoma. AB - Chromosomal deletions at segment 11q23-q24 have been identified in a variety of human epithelial tumors, including cervical carcinoma (CC), indicating the presence in this region of at least a tumor suppressor gene (TSG) involved in the development of these neoplasms. To localize the 11q deletion target more precisely, 54 primary cervical carcinomas were examined for loss of heterozygosity (LOH) using a panel of microsatellite DNA markers mapping to 11p.15 and spanning region 11q23-qter. Nineteen tumors were found to have LOH at chromosome 11q. The highest frequency of LOH was observed at locus APOC-3, located in 11q23.1-q23.2, which was deleted in 42% of the informative cases. In contrast, LOH was infrequent at distal 11q in current series of CC. The smallest common region of loss included APOC-3 and was defined distally by marker D11S925 in region 11q23. The present data strongly suggest that the 11q suppressor gene(s) involved in cervical tumorigenesis is likely to be located at chromosome region 11q22-q23. PMID- 10475384 TI - Rapid test for identification of a human papillomavirus 16 E6 L83V variant. AB - A variant of human papillomavirus (HPV) 16 has been shown recently to be more prevalent in invasive cervical carcinoma than in preinvasive lesions. This HPV 16 variant possesses a common mutation (T to G) in nucleotide 350 (codon 83) of the E6 gene, resulting in an amino acid shift, L83V, in the E6 protein. This mutation was believed to signify preinvasive cervical lesions with a high probability of progression to invasive carcinoma. The purpose of the present investigation is to describe a rapid method for the detection of this variant HPV 16, E6 (L83V). Paraffin blocks of 18 gynecologic biopsy specimens were collected, all displaying the morphology of cervical intraepithelial neoplasia (CIN I-III) and a positive HPV 16 test. Sections from these blocks were used for DNA extraction. A DNA sequence of the E6 gene containing 176 bases (including codon 83) was amplified by polymerase chain reaction (PCR) and analyzed by non-radioactive single-strand conformational polymorphism (SSCP) analysis. A divergent SSCP pattern was observed in 7 of the HPV 16 positive biopsy specimens. A DNA sequence analysis of the PCR products revealed the conversion of Leu to Val in codon 83 of the E6 gene, correlating to the divergent band pattern. This PCR-SSCP method can be used to test for HPV 16 in women who are at serious risk of developing invasive cervical carcinoma. PMID- 10475385 TI - Processed pseudogene from the von Hippel-Lindau disease gene is located on human chromosome 1. AB - The von Hippel-Lindau (VHL) disease gene is a tumor suppressor located at 3p25 26. While amplifying intron 1 of this gene, a smaller-than-expected product was found. This fragment was sequenced and was approximately 78% similar in sequence to the VHL gene and completely lacked sequence from the intron. No stop codons were found in the sequenced region. Using this DNA fragment as a probe for Northern blot hybridization analysis, no evidence was found for expression of a unique RNA. Because of the lack of intron 1 sequence and the likely lack of expression, the new sequence is most probably a part of a VHL processed pseudogene. The putative pseudogene was mapped to human chromosome band 1q12 using the polymerase chain reaction with template DNA from human/rodent somatic cell hybrids, a radiation hybrid panel, and a set of primers that were chosen to be maximally divergent from the genuine VHL gene. The human/rodent somatic cell hybrid DNAs were then used on Southern blots to determine which human bands are from the pseudogene and which are from the functional gene. This knowledge is valuable in interpreting Southern blot evidence of VHL gene abnormalities. PMID- 10475386 TI - Thoracic Surgery Directors Association Award. Angiogenesis in transmyocardial revascularization: comparison of laser versus mechanical punctures. AB - BACKGROUND: Transmyocardial laser revascularization (TMLR), which has been shown to reduce angina in clinical trials, was originally based on the belief that laser channels are unique and can remain patent. An increasing body of evidence indicates otherwise, and transmyocardial revascularization (TMR) angiogenesis is currently thought to be induced by nonspecific inflammatory response to tissue injuries. We tested the hypothesis that mechanical transmyocardial revascularization (TMMR) may induce angiogenic responses similar to that seen with lasers. METHODS: Ameroid constrictors were implanted around proximal circumflex arteries of porcine hearts. Six weeks later, they were randomly assigned (n = 5 each) to receive 10 transmural channels in the ischemic zone by a carbon dioxide laser (group I) or by a needle (group II). A third group (group III) had 30 needle channels in the same area, while a control group (group IV) received no TMR. The hearts were harvested 1 week later, and, using immunohistochemistry, vascular endothelial growth factor (VEGF) expression was studied and quantified by computerized morphometric analysis. Densities of vascular structures positively stained for VEGF per high-power field (HPF) were also compared. RESULTS: Virtually no TMR channels remained patent histologically. Group III had a significant higher level of total VEGF expression (14.18+/-0.78 mm2) compared with group I (7.07+/-2.06 mm2, p < 0.001) and group II (4.74+/-3.35 mm2, p < 0.001). Vascular density was significantly elevated in all treatment groups compared with the control (group I, 7.7+/-0.8/HPF vs group II, 4.5+/ 2.3/HPF vs group III, 8.1+/-0.6/HPF vs group IV, 1.1+/-0.5/HPF). CONCLUSIONS: In view of the significant cost implications, our findings that needle punctures may also induce angiogenic response comparable with that with laser suggest that it is justifiable and desirable to include a TMMR arm for comparison with TMLR in future clinical trials. PMID- 10475387 TI - Surgeons' assessment of symptoms suggesting extrathoracic metastases in patients with lung cancer. Canadian Lung Oncology Group. AB - BACKGROUND: In patients with apparently operable non-small cell lung cancer (NSCLC), clinicians often omit investigation for M disease in asymptomatic patients. Previous investigations have not specified in detail what is meant by "symptomatic," and this could differ between surgeons. We have investigated the extent to which surgeons' criteria differ for presence of symptoms. METHODS: Participating surgeons from seven centers, enrolled patients they judged "asymptomatic" in a randomized trial of investigational strategies for NSCLC. Patients completed a structured questionnaire describing symptoms of the central nervous system (CNS). In 685 patients, we documented CNS symptom recurrence after resectional surgery over 1 year of follow-up. RESULTS: Two centers enrolled only patients without even the mildest symptoms. Three centers took an intermediate approach, occasionally classifying patients with mild symptoms as "asymptomatic" and thus enrolling them in the trial. Two centers classified an appreciable number of patients with minimal symptoms, and occasionally with more than minimal symptoms, as "asymptomatic." Patients with even mild CNS symptoms were more likely to subsequently present with CNS metastases. CONCLUSIONS: Thoracic surgeons differ in their ideas of what may constitute the symptoms of M disease. Patients with structured questionnaire results that suggest symptoms of CNS disease are more likely to have CNS symptom recurrence after resectional surgery. PMID- 10475388 TI - Safe pulmonary resection after chemotherapy and high-dose thoracic radiation. AB - BACKGROUND: Pulmonary resection after high-dose thoracic irradiation is reported to be associated with a high morbidity and mortality, and has been considered to be prohibitive. METHODS: We report safe pulmonary resection in 19 consecutive patients receiving neoadjuvant therapy that included greater than 59 Gy thoracic radiation. The mean thoracic radiation dose was 61.8 Gy (range 59.5-66.5) and mean age was 52 years (range 36-72 years). Cell type was adenocarcinoma (6), squamous (7), and other non-small cell lung cancer (NSCLC) (6). Sixteen of 19 patients received concurrent chemotherapy. Median time from end of treatment to surgical resection was 89 days (range 22-258 days). Surgical resection included 13 lobectomies and six pneumonectomies (four right, two left). RESULTS: A complete pathologic response was seen in 8 of 19 (42%) patients. Three patients required intraoperative transfusion of blood. Mean intensive care unit stay was 2.0 days (range 1-8 days), and mean length of stay (LOS) was 8.0 days (range 3-18 days). There were four postoperative complications; one bronchopulmonary fistula, one subarachnoid-pleural fistula, and 2 patients with prolonged atelectasis. There was no incidence of acute respiratory distress syndrome (ARDS) or operative mortality. CONCLUSIONS: Pulmonary resection, including pneumonectomy, after chemotherapy and high-dose thoracic radiation may be performed safely with a low rate of intraoperative and postoperative complications. PMID- 10475389 TI - Surgical aspects of resection for suspected invasive pulmonary fungal infection in neutropenic patients. AB - BACKGROUND: Morbidity and mortality of early resection of invasive pulmonary fungal disease in neutropenic patients are still considered prohibitive for surgical treatment. METHODS: We retrospectively analyzed results of 28 (16 men, 12 women; mean age, 38.9 years) consecutive neutropenic hematologic patients who had lung resections for suspicion of invasive pulmonary fungal disease. RESULTS: We did 28 lung resections (19 lobectomies, one bilobectomy, eight single or multiple wedge resections including three video-assisted wedge resections). The disease was proved histologically in 22 (78.6%) cases. Intraoperative difficulties, such as diffuse oozing or mycotic infiltration, and solid postinflammatory adhesions were encountered in 5 (17.8%) and 6 (21.4%) patients respectively. In one case (3.6%) it lead to a major intraoperative hemorrhage. There were no intraoperative deaths, overall 30-day mortality rate was two of 28 (7.1%), overall 90-day mortality rate was seven of 28 (25%), with one death (3.6%) possibly related to surgery. Minor surgery-related complications were seen in ten (35.7%) cases, major surgery-related complications occurred in three (10.7%) cases. Twelve of 22 patients (54.5%) with proven invasive fungal infection are currently alive (mean follow-up, 32.3 months). CONCLUSIONS: Surgery related complications and mortality are acceptable for this high risk group of patients. Resection should be carried out early for diagnostic as well as therapeutic reasons. PMID- 10475390 TI - Evaluation of TMN classification for lung carcinoma with ipsilateral intrapulmonary metastasis. AB - BACKGROUND: Staging for lung cancer based on the TNM classification is an important predictive factor for prognosis. Recently, lung cancer with ipsilateral intrapulmonary metastasis (PM) was reclassified according to the revision of the TNM classification. To evaluate the prognostic importance of the new staging system for PM, we analyzed the postoperative survival of patients with non-small cell lung carcinoma. METHODS: Of 1,002 consecutive patients who underwent operation for primary lung cancer between June 1984 and December 1996, we reviewed the medical record of 889 patients who underwent complete resection for non-small cell lung cancer. RESULTS: We considered 89 patients (10.0%) to have synchronous ipsilateral PM. After reclassification to the former staging system revised in 1992, 5 patients were classified as stage I, 29 as stage IIIA, 48 as stage IIIB, and 7 as stage IV. In the new staging system revised in 1997, 48 patients were recategorized as stage IIIB, and 41 as stage IV. The 5-year survival of patients without PM (49.5%) was significantly better than that of patients with PM in primary-tumor lobe (29.6%, p = 0.002) or in nonprimary-tumor ipsilateral lobe (23.4%, p = 0.0002). Although the survival of patients with stage IV cancer without PM was significantly worse than that of patients with the new (1997) stage IV cancer with PM (p = 0.02), it was similar to that of patients with the former (1992) stage IV cancer with PM. The survival of PM patients with N0 or N1 disease was significantly better than that of PM patients with N2 or N3 disease (p = 0.001). Furthermore, in patients with the new (1997) stage IIIB cancer, the survival of N0 disease was better than that of N2 disease (p = 0.007). CONCLUSIONS: Inasmuch as the prognosis of non-small cell carcinoma in patients with PM strongly correlated with N factor rather than PM factor, N factor should be reflected in a staging designation. We therefore consider the new TNM classification for PM reclassified in 1997 to be less acceptable for surgical-pathologic staging than the revision in 1992. PMID- 10475391 TI - Anomalous systemic arterial supply to normal basal segments of the left lower lobe. AB - BACKGROUND: Anomalous arterial supply to the normal basal segments of the lower lobe without sequestration is a rare congenital abnormality, and whether it belongs to the broad spectrum of sequestration disorders remains controversial. METHODS: The cases of all 4 patients who were treated surgically by us were reviewed together with 8 previously reported cases. RESULTS: The anomalous artery originated from the descending thoracic aorta, distributed to the basal segments of the left lower lobe, and drained to a normal inferior pulmonary vein in each case. The anomalous artery was thick and elastic walled. From the review of all 12 cases, male gender, left side, descending thoracic aorta as the aberrant arterial origin, absence of pulmonary blood flow to the basal segments, and normal pulmonary venous drainage were predominant. Despite some differences, the findings seemed closely related to intralobar sequestration. Surgical treatments were lung resection, anastomosis, and ligation of the anomalous artery. CONCLUSIONS: This anomaly is probably one type of sequestration complex. Both aortic and pulmonary arterial angiographic studies are needed to plan the definitive surgical procedure. PMID- 10475392 TI - Prevention of postoperative air leakage from lungs using a purified human collagen membrane-polyglycolic acid sheet. AB - BACKGROUND: Human amnion has useful biomedical applications because it contains a large amount of human collagen fibers. We prepared purified human collagen membrane (HCM) from human amnion and used it to develop a new sheet by combining it with synthetic bioabsorbable polyglycolic acid (PGA) mesh. We evaluated its efficacy in preventing air leakage from the lungs of dogs. METHODS: In 20 dogs, HCM-PGA sheet (n = 5), sheets using fibrin glue with a separate application method (n = 5), a mixed application method (n = 5), and fibrin glue alone (n = 5), were used as dressing materials after partial lung resection. RESULTS: The HCM-PGA sheet using fibrin glue with a separate application method was shown to be significantly more effective by an air leakage pressure test than the other three methods. These results indicate that the HCM-PGA sheet is useful for preventing air leakage from the lung. CONCLUSIONS: The HCM-PGA sheet is more effective than conventional fibrin glue for controlling postoperative air leakage. PMID- 10475394 TI - Accountability: the future application of cardiothoracic surgical data in a changing health care environment. PMID- 10475393 TI - Significance of P53 and Rb protein expression in surgically treated non-small cell lung cancers. AB - BACKGROUND: Alterations of the P53 or Rb gene are among the most frequently observed genetic changes in primary lung cancer. Nevertheless, there has been no final conclusion on the relationship between P53 or Rb protein expression and clinico-pathological parameters in primary lung cancer. A large-scale study was performed to examine the clinicopathological and prognostic significance of P53 and Rb expressions in 207 surgically resected non-small cell lung cancer (NSCLC) patients. METHODS: Tumor specimens were obtained from 207 primary NSCLC surgically resected from January 1990 through December 1994. The avidin-biotin peroxidase method was used to determine the expression of P53 or Rb of tumor cells using anti-P53 or anti-Rb monoclonal antibodies. The relationships between P53 or Rb protein expression and the clinicopathological parameters were analyzed. RESULTS: Expression of P53 or Rb protein was detected in 115 (55.6%) and 136 (65.7%) of the 207 lung tumors, respectively. P53 had significantly higher positive results in patients with regional lymph node metastasis and advanced tumor stage. Rb expression was significantly lower in lung cancers with a macro- or microscopic picture of tumor necrosis. Additionally, an inverse correlation between the expression of Rb and P53 was found. By multiple variate analysis, P53 expression and pathological stage were independent, significant prognostic factors. Further analysis demonstrated P53 expression was an independent prognostic factor in stage 1, but not in other stages. CONCLUSIONS: P53 expression is especially useful as a prognostic factor in stage 1 lung cancer. PMID- 10475395 TI - Using database information in your clinical practice. AB - BACKGROUND: Clinical database information is in wide demand, but it is not always used to its full potential. Clinicians must learn to be the experts and to assert leadership in the use of their own data. METHODS: Clinical databases provide unique perspectives on the full process of care for a heterogeneous population of patients. They focus beyond individual providers, to their interaction within a complex system of care. Exploring questions that arise from this data can identify system issues, which are invisible to individual practitioners or specialties using other traditional review methods. RESULTS: Clinical database information helped our hospital staff identify problems in their approach to a small subset of high risk coronary bypass patients. Multiple system changes resulted in a dramatic reduction in mortality. Collateral impact on all care reduced overall mortality from 4.5% to below 2%. CONCLUSIONS: The greatest opportunities for improvement in patient care often lie in the areas where specialties or teams interface, eg, in overlapping or transferring care. The whole system of care for each patient must be optimized, not just the individual specialty components. Clinical database information provides a way to evaluate and improve the overall process and results of the broader system of patient care. PMID- 10475396 TI - Measuring for improvement: from Toyota to thoracic surgery. AB - BACKGROUND: Measuring quality has become a high priority in the era of managed care. Nevertheless, it can be counterproductive to use the same methods for measuring improvement in surgical procedures and processes as we use for measurement in basic research. Techniques of statistical process control have been used for many years to measure process improvement in industry and are now being applied to health care. METHODS: Examples of using statistical process control charts to monitor coronary artery bypass grafting mortality, intensive care unit admission time, and length of stay are reviewed. RESULTS: The major advantage of using control chart methodology is that it allows one to determine whether the process being evaluated is in fact stable and to detect when significant or special cause variation occurs. CONCLUSIONS: Summary statistics currently provided to purchasers of care and regulatory agencies do not ensure that the processes being evaluated are stable. We need to look at data over time with statistically validated methods such as control charts to better monitor our processes of care and thereby provide accurate statistics. PMID- 10475397 TI - Innovative uses of a cardiothoracic database. AB - This report describes the activities of the Northern New England Cardiovascular Disease Study Group. The group consists of representatives from six institutions: Eastern Maine Medical Center in Bangor, and Maine Medical Center in Portland, ME; Optima Health Care in Manchester and Dartmouth-Hitchcock Medical Center in Lebanon, NH; Fletcher Allen Health Care in Burlington, VT; and Beth Israel Deaconess Medical Center in Boston, MA. The Northern New England Cardiovascular Disease Study Group maintains a voluntary primary cardiac surgical database that has risk-stratified information on more than 60,000 consecutive patients who have undergone open heart surgical procedures in northern New England since 1987. In 1991, the group reported that significant variation in mortality rate existed between centers, a difference not explained by case mix. The finding led to a regional retrospective review of deaths in an effort to identify "mode of death." Mode of death is defined as that event that started the chain of events ultimately leading to the death of the patient. The most common mode of death was found to be low cardiac output syndrome. This information has led to a regional effort toward prevention, early recognition, and successful treatment of low cardiac output syndrome in the perioperative period. PMID- 10475398 TI - Closing the loop: optimizing physicians' operational and strategic behavior. AB - Clinical databases are essential elements in optimizing medical care. They are no finality by themselves, but essential elements in the generation of knowledge. Optimal medical care starts with optimal care based on existing knowledge. This care continues with the registration of the variability in morbidity, comorbidity, and therapy, but also the registration of the early and late outcome. This should then allow the generation of structured inferences based on this registration and the closure of the loop, by treating patients according to this newly created evidence. PMID- 10475399 TI - The Society of Thoracic Surgeons National Database: current status and future directions. AB - The Society of Thoracic Surgeons National Database, established seven years ago by thoracic surgeons for self improvement and quality assurance, now has 1,181,464 patients registered, including 897,914 coronary artery bypass operations. Risk-adjusted 30 day mortality for coronary bypass procedures, unadjusted mortality for other cardiothoracic procedures, unadjusted morbidity and length of stay as well as several processes of care are measured. There has been a progressive decrease in operative mortality and length of stay over the past seven years. Deaths, complications, and lengths of stay are stratified according to estimated risk of death. Definitions have been refined in conjunction with the American College of Cardiology. The database is being increasingly utilized for state analyses and is in demand by other organizations and third party carriers. Logistic regression analysis is now utilized for development of the risk models. The database has been useful for health care policy decisions and can be useful for our Professional Affairs Committee in their dealings with government. Other uses include measuring access to care and cost. Data quality improvement measures have been put in place, as well as data manager education. The General Thoracic and Congenital data acquisition packages are being modified and improved, and a goal is to begin collecting longitudinal data to demonstrate the long term efficacy of thoracic procedures. The data elements have been decreased from 500 to 200+ core variables for simplification. With the changing healthcare environment and emphasis on cost cutting, collecting valid data by a national specialty group enhances the monitoring of quality of care, thus protecting our patients from overzealous cutbacks. Data is essential to document the efficacy, quality and cost-effectiveness of the procedures we perform and is a necessary tool for each of us to have to assure the quality and continued success of our practices. PMID- 10475400 TI - An essential role for NF-kappaB in the cardioadaptive response to ischemia. AB - BACKGROUND: Ischemic preconditioning (IP) is the phenomenon whereby brief episodes of ischemia protect the heart against a subsequent ischemic stress. We hypothesize that activation of the transcription factor NF-kappaB mediates IP. METHODS: Rabbits were randomly allocated to one of three groups: (1) 45 minutes of myocardial ischemia followed by 2 hours of reperfusion (I/R); (2) three cycles of 5-minute ischemia and 5 minutes of reperfusion followed by I/R (IP + I/R); or (3) IP in the presence of ProDTC, a specific NF-kappaB inhibitor, followed by I/R (IPProDTC + I/R). Infarct size, indices of regional contractility, and NF-kappaB activation were determined. RESULTS: In preconditioned rabbits (IP + I/R), infarct size was reduced 83% compared with both I/R alone and IPProDTC + I/R groups (p < 0.05). Throughout reperfusion, preconditioned myocardium showed enhanced regional contractile function compared with I/R and IPProDTC + I/R groups (p < 0.05). Gel shift analysis showed NF-kappaB activation with IP that was blocked by ProDTC. I/R and IPProDTC + I/R groups showed NF-kappaB activation with I/R that was absent in preconditioned animals. CONCLUSIONS: The cytoprotective effects induced by IP require activation of NF-kappaB. PMID- 10475401 TI - Results of graft patency by immediate angiography in minimally invasive coronary artery surgery. AB - BACKGROUND: Although minimally invasive direct coronary artery bypass (MIDCAB) is being employed for revascularization of the left anterior descending coronary artery (LAD) with the left internal mammary artery (LIMA), little objective data exist regarding graft patency. Because the procedure is performed on a beating heart through a limited access approach, concerns have been raised regarding the ability to perform as accurate an anastomosis compared with conventional coronary artery bypass (CAB). METHODS: A prospective study of consecutive patients undergoing MIDCAB LIMA to LAD was undertaken. All procedures were performed through a limited anterior thoracotomy incision with a stabilization device. Selective angiography of the LIMA graft was performed intraoperatively or in the immediate postoperative period. RESULTS: One hundred and three patients underwent the MICAB procedure. Angiographic evaluation of the anastomosis was obtained in 100 patients (97%). Angiographic graft patency was 99%, with perfect graft patency (no stenosis greater than 50%) being 91%. Three grafts were revised in the operating room. One patient underwent reoperation and 3 more underwent percutaneous transluminal coronary angioplasty. There were two noncardiac mortalities (1.9%), both with patent grafts. CONCLUSIONS: Immediate graft patency after MIDCAB is acceptable, and comparable with conventional CAB data, although meaningful comparison is difficult. The significance of early angiographic findings and the role for early angiography remain to be defined. PMID- 10475402 TI - Stroke in cardiac surgical patients: determinants and outcome. AB - BACKGROUND: Despite improving outcomes in cardiac surgical patients, stroke continues to remain a major complication. Few prospective studies are available on postoperative stroke. The present study was conducted to elucidate the incidence and predictors of stroke in a large group of cardiac surgical patients. METHODS AND RESULTS: Prospective data collected on 4,941 patients undergoing cardiac surgery were subjected to univariate and logistic regression analyses (98.4% men; 72% older than 60 years; 9.1% with history of prior stroke; 80.4% underwent isolated coronary artery bypass grafting). Stroke predictors include history of stroke and hypertension, older age, systolic hypertension, bronchodilator and diuretic use, high serum creatinine, surgical priority, great vessel repair, use of inotropic agents after cardiopulmonary bypass, and total cardiopulmonary bypass time (p < 0.05 for all comparisons). Median intensive care unit and hospital stays were longer, and hospital mortality and 6-month mortality were higher for patients with stroke (p < 0.001). CONCLUSIONS: Stroke after cardiac surgical procedures is a morbid event. Identification of predictors and development of strategies to modify these factors should lead to a lower incidence of stroke. PMID- 10475403 TI - Total arterial revascularization with an internal thoracic artery and radial artery T graft. AB - BACKGROUND: Proximal anastomosis of the radial artery to the side of the internal thoracic artery (ITA) permits complete arterial revascularization in most patients, with the aim of improving long-term results of coronary artery bypass through greater long-term graft patency. The short-term results, however, have yet to be defined. We therefore reviewed our early experience with this grafting strategy. METHODS: Between October 1, 1993, and September 1, 1998, 649 patients aged 30 to 85 years (mean, 60+/-10 years) had primary coronary artery bypass using an ITA and radial artery in a T-graft configuration. Left ventricular function was severely depressed (ejection fraction <35%) in 12%, and left main stenosis was present in 14%. RESULTS: A total of 937 distal anastomoses were performed with the left ITA (1.4 per patient) and 1,452 with the radial artery (2.2 per patient). There was one perioperative death (0.2%). There were 32 (5%) q wave myocardial infarctions, and 14 patients (2%) had transient low output syndrome. There was one episode of hypoperfusion corrected by lengthening the left ITA. Angiography for clinical indications in 27 patients 1 to 35 months postoperatively (mean, 9.5+/-8.3 months) demonstrated a distal anastomotic patency of 100% for ITA and 82% for radial artery grafts. CONCLUSIONS: Complete arterial revascularization can be achieved with an ITA and radial artery T-graft with low operative risk and acceptable early patency. These results support the continued investigation of this grafting strategy. PMID- 10475404 TI - Routine use of bilateral skeletonized internal mammary arteries for myocardial revascularization. AB - BACKGROUND: Complete arterial myocardial revascularization without the use of saphenous veins grafts was primarily performed on selected patient populations such as the young and nondiabetic. In a recently developed surgical technique, the internal mammary artery is dissected gently as a longer skeletonized artery, providing greater versatility for complete arterial revascularization, without saphenous veins grafts. METHODS: We prospectively evaluated the impact of the routine use of double skeletonized internal mammary artery in 472 patients who underwent coronary artery bypass grafting between April 1996 and June 1997. Their average age was 65 years (30 to 87 years), 383 (83%) were men, and 89 (17%) women. One hundred sixty-nine (36%) of the patients were older than 70 years, and 145 (31%) were diabetic. The average number of grafts was 3.2 per patient (two to six grafts). RESULTS: Operative mortality was 1.7% (n = 8). The mortality of urgent and elective patients was 0.7% (3 of 410 patients), and that of emergency operations was 8.1% (5 of 62 patients; p < 0.01). There were three (0.6%) perioperative infarcts, and 6 patients (1.3%) sustained strokes. Sternal wound infection occurred in 8 patients (1.7%). Postoperative follow-up (1 to 25 months) was available in 462 patients (99%). Two-year actuarial survival was 96.8%, and 92% of the surviving patients are well and free of angina. Neither diabetes mellitus nor old age (>70 years) were significant independent predictors of any early or late untoward events. None of the 70 diabetic patients more than 65 years of age developed sternal wound infection. Chronic lung disease was found to be the only independent predictor for sternal infections. CONCLUSIONS: Routine use of bilateral skeletonized internal mammary artery is a safe replacement for the current myocardial revascularization technique even in the old and diabetic patients. PMID- 10475405 TI - Nitecapone as an additive to crystalloid cardioplegia in patients who had coronary artery bypass grafting. AB - BACKGROUND: Nitecapone has been shown to have a protective effect against ischemia-reperfusion injury in experimental heart transplantation and in Langendorff preparations. This prospective, randomized study assessed the effects of nitecapone in patients who had coronary artery bypass grafting. METHODS: Thirty patients with normal myocardial function were randomly divided into control patients (n = 15), who received crystalloid (Plegisol) cardioplegia, and nitecapone patients, who received nitecapone in a 50 microM solution (n = 15) in Plegisol. Cardioplegia was administered as an initial dose of 15 mL/kg of body mass after cross-clamping and 2 mL/kg every 15 minutes. Simultaneous coronary sinus and aortic blood samples, and myocardial biopsies were taken at 1, 5, and 10 minutes after unclamping. Hemodynamics were measured invasively for 24 hours and with transesophageal echocardiography for 3 hours after cardiopulmonary bypass. RESULTS: There were no adverse effects. The incidence of ventricular arrhythmias was significantly lower in the treatment group during the recovery period (p = 0.02). Cardiac output and stroke volume did not differ significantly between the groups. The conjugated dienes gradient between the aorta and the coronary sinus increased significantly during the first minute of reperfusion in the control group (p = 0.02) compared with the nitecapone group. Myeloperoxidase activity in myocardial biopsies was higher in the control group (2.3 times higher at 5 minutes and 3.2 times higher at 10 minutes) than in the nitecapone group (p = 0.13). CONCLUSIONS: Nitecapone did not exert any significant hemodynamic effects in patients with normal ejection fraction. PMID- 10475406 TI - Aortic Carpentier-Edwards supraannular porcine bioprosthesis: a 12-year experience. AB - BACKGROUND: After 35 years of cardiac valve replacement, the ideal substitute remains to be found. Homografts are considered best but, due to their scarcity, cannot meet the need of valve replacement. Artificial valves (mechanical or biological) remain the most commonly used but controversy is still present as to the better choice. We tested the Carpentier-Edwards bioprosthesis for its efficacy in valve replacement operations. METHODS: From 1983 to 1995, 1,108 consecutive patients had an isolated aortic valve replacement with a porcine Carpentier-Edwards bioprosthesis, model 2650 supraannular valve. Mean age was 73.8+/-8.3 years. Aortic stenosis was the most common lesion (1,049 patients, 94.7%). The follow-up of 980 operative survivors was 96% complete and represented a total of 4,735 patient-years (maximum, 13.8 years; mean, 4 years and 10 months). RESULTS: Actuarial survival including operative mortality (128 patients, 11.6%) was 43.6%+/-2.3% at 10 years and 27.3%+/-3.3% at 12 years and, at that time, was not statistically different from those of the normal French population matched for age and sex. Structural deterioration of the valve was observed in 27 patients, an actuarial freedom of 94.2%+/-1.5% at 10 years and 83.8%+/-4.5% at 12 years. Hazard function revealed a stable and low risk of structural deterioration until 10 years and significantly increased risk after that. Young age was found to be an increasing risk factor of deterioration. Reoperation for valve-related complications was necessary in 30 patients, an actuarial freedom of 94.5%+/-1.4% at 10 years. CONCLUSIONS: The Carpentier-Edwards porcine supraannular valve affords a good durability up to 10 years, with a low rate of reoperation. The risk of structural deterioration decreases with older age. It is our valve of choice in elderly patients. PMID- 10475407 TI - Treatment of moderate mitral regurgitation and coronary disease by coronary bypass alone: late results. AB - BACKGROUND: In cases of moderate mitral regurgitation and coronary artery disease operative strategy continues to be debated between coronary artery bypass grafting alone and concomitant valve replacement or repair. We previously reported on 58 patients with moderate mitral regurgitation who had coronary artery bypass grafting between 1977 and 1983. We present the late results for this original cohort (test group), and a matched control group of coronary artery bypass grafting patients without mitral regurgitation (n = 58). METHODS AND RESULTS: In the original cohort, the hospital mortality rate was 3.4% (2 of 58), and 80.4% (45 of 56) of hospital survivors were alive at the time of initial follow-up (mean, 4.3+/-2.3 years). Hospital mortality in the control group was 6.9% (4 of 58 patients). Follow-up was 98.2% (108 of 110 patients) complete, with a mean follow-up time of 10.3+/-5.5 years. Kaplan-Meier curves for hospital survivors showed similar 5- and 10-year survival rates between the two groups (p = 0.59). On multivariate analysis, age 65 years or more, congestive heart failure class III or IV, and pulmonary capillary wedge pressure more than 17 mm Hg were significant (p < 0.05) independent predictors of diminished survival in the test group. CONCLUSIONS: Patients with moderate mitral regurgitation and coronary artery disease treated solely with coronary artery bypass grafting had acceptable early and late results. Moderate mitral regurgitation at the time of revascularization does not always warrant operative correction. PMID- 10475408 TI - Initial experience with MIDCAB grafting using the gastroepiploic artery. AB - BACKGROUND: Minimally invasive direct coronary artery bypass grafting with the gastroepiploic artery can be used in primary operations and reoperations to revascularize the inferior or anterior surface of the heart. METHODS: Patients who had symptomatic coronary artery disease limited to a single coronary distribution were selected. Coronary targets were grafted with the pedicled gastroepiploic artery through a small midline epigastric incision. Patients were followed with scheduled outpatient clinic visits, Doppler examination, and selective recatheterization. RESULTS: Between May 1995 and November 1997, 74 patients underwent gastroepiploic artery minimally invasive direct coronary artery bypass grafting; 33 (45%) had a primary operation and 41 (55%), a reoperation. Grafting was performed to the distal right coronary artery (n = 38), the posterior descending artery (n = 28), or the distal left anterior descending coronary artery (n = 8). There were six deaths (8%) within 30 days after operation. Twenty patients (28%) underwent recatheterization; there were two graft occlusions, two graft stenoses, and five anastomotic stenoses. Of 60 patients seen 2 or more weeks after operation, 53 (88%) had resolution of anginal symptoms at a mean follow-up of 10.9 months (range, 0 to 30 months). CONCLUSIONS: Inferior minimally invasive direct coronary artery bypass grafting with the gastroepiploic artery avoids the risks of repeat sternotomy, aortic manipulation, and cardiopulmonary bypass. Patency rates, however, were lower than expected, and there is significant morbidity and mortality associated with high-risk patients undergoing the procedure. Continued follow-up is essential to evaluate long-term graft patency and patient survival. PMID- 10475409 TI - Fast track recovery of elderly coronary bypass surgery patients. AB - BACKGROUND: To ascertain whether early extubation and fast-track treatment protocols are feasible in elderly patients, we analyzed 487 consecutive patients who had isolated coronary artery bypass grafting between January 1995 and June 1997, constituting the experience of a single surgeon. METHODS: Management consistently applied to all patients emphasized early extubation protocol, tepid cardioplegia and normothermic bypass to reduce pump times, early mobilization and chest tube removal, and protocol treatment of atrial fibrillation. Elderly patients at least 70 years old (n = 176, mean age 75 years) were compared with younger patients (n = 311, mean age 58 years). RESULTS: The hospital mortality rate was 0.8% (4 of 487 patients), and there was no difference in the operative mortality rate of the older cohort versus the younger cohort (0.6% versus 0.9%; p > 0.05). Older patients had a higher incidence of peripheral vascular disease, congestive heart failure, prior strokes, renal failure, and cerebrovascular disease (p < 0.05). Early extubation was achieved in 71% of younger patients versus 57% of older cohort (95% confidence interval, 14%+/-9%; p = 0.002). Older patients had significantly higher incidence of postoperative atrial fibrillation (27% versus 14%; 95% CI, 13%+/-7%; p < 0.001), a factor responsible for shorter length of stay among younger patients (5.6+/-2.8 days versus 7.2+/-3.7 days; 95% CI, 1.6+/-0.3 days; p < 0.001). Nonetheless discharge before the fifth postoperative day was achieved in 34% of the elderly patients. CONCLUSIONS: Although elderly patients have a higher acuity of illness, critical pathways for accelerated discharge are safe and feasible in most elderly patients. PMID- 10475410 TI - Influence of polymorphonuclear leukocytes and plasma on coronary vasomotion after ischemia. AB - BACKGROUND: The role of plasma and neutrophils in endothelial dysfunction following myocardial ischemia was investigated. METHODS: Isolated rabbit hearts were perfused at constant pressure with a modified Krebs-Henseleit solution. After a 30-minute global ischemia the hearts were perfused at the onset of reperfusion for 10 minutes with either NaCl (group I, n = 6), autologous plasma alone (group II, n = 5), autologous polymorphonuclear leukocytes alone (PMN, group III, n = 6), or PMN and plasma in combination (group IV, n = 5). Before and after ischemia the effects of intracoronary endothelial dependent and independent vasodilation by acetylcholine (1 x 10(-7)mol/L) and glycerol trinitrate (1 x 10( 6) mol/L) were investigated. RESULTS: A similar increase in coronary flow was induced in groups I, II, and III by acetylcholine and glycerol trinitrate before and after ischemia. In contrast, in group IV the endothelial dependent vasodilation was significantly depressed (p < 0.05). In groups II and IV a moderate but significant reduction in the recovery of the left ventricular pressure was observed after ischemia and reperfusion. CONCLUSIONS: These results suggest that after myocardial ischemia, plasma is required for neutrophil mediated endothelial dysfunction. PMID- 10475412 TI - Warm or cold continuous blood cardioplegia provides similar myocardial protection. AB - BACKGROUND: This study was performed to investigate the effect of temperature of blood cardioplegia on the recovery of postischemic cardiac function. METHODS: Pigs on cardiopulmonary bypass were subjected to global ischemia (30 minutes), followed by cold (n = 10) or warm (n = 11) continuous antegrade blood cardioplegia (45 minutes) delivered at 55-60 mm Hg. RESULTS: Global left ventricular function, evaluated by preload recruitable stroke work, decreased with cold cardioplegia from 91 (85-103) [mean (quartile interval)], at baseline, to 73 (55-87) erg x 10(3)/mL postbypass (p = 0.03), but was unchanged after warm cardioplegia; 110 (80-132) to 109 (71-175) erg x 10(3)/mL (p > 0.5). However, the difference between treatment effects was not significant (p = 0.25). Diastolic function, evaluated by end-diastolic pressure-volume relation, deteriorated without any difference between groups. Mean cardioplegic flow was similar between groups. Coronary vascular resistance increased at constant rate during warm cardioplegic delivery, but remained unchanged with cold cardioplegia (p = 0.001 between regression coefficients). CONCLUSIONS: No significant difference was found in postischemic functional recovery comparing cold and warm continuous blood cardioplegia. Cold cardioplegia is therefore preferred due to added safety of hypothermia. PMID- 10475411 TI - Antegrade crystalloid cardioplegia vs antegrade/retrograde cold and tepid blood cardioplegia in CABG. AB - BACKGROUND: This study evaluated the myocardial protective strategies in isolated coronary bypass surgeries. METHODS: One hundred and twenty-eight patients were prospectively randomized to 3 techniques of myocardial protection; group I (n = 47) antegrade/retrograde tepid blood cardioplegia, group II (n = 40) antegrade/retrograde cold blood cardioplegia with topical cooling, group III (n = 41) antegrade crystalloid cardioplegia with topical cooling. RESULTS: The incidence of spontaneous defibrillation was significantly higher in group I (p < 0.001) while the incidence of low cardiac output was not different between the 3 groups. The incidence of ventricular arrhythmia was higher in group III (p < 0.016 group III vs I). There was no significant statistical difference in hemodynamic recovery between the 3 groups. CK-MB levels were significantly lower in group I versus the other 2 groups, (p = 0.0013, 0.04). Acid release and oxygen extraction were higher in group II than in group I (p = 0.06) during cardioplegia and reperfusion. Lactate release was less in group I at the release of aortic cross-clamp, and reperfusion. There was no significant difference between the 3 groups in ICU stay, ventilation time, or hospital complications. CONCLUSIONS: Tepid blood cardioplegia showed superiority in metabolic and functional recovery, whereas crystalloid cardioplegia had the highest incidence of postoperative arrhythmias. There was no significant statistical difference between the 3 groups in hospital mortality and morbidity. PMID- 10475413 TI - Transmyocardial laser revascularization does not denervate the canine heart. AB - BACKGROUND: Transmyocardial laser revascularization has been used as an indirect approach to improve myocardial perfusion in patients with refractory angina not amenable to conventional therapy. Three mechanisms have been proposed for its therapeutic effects: direct perfusion of the ischemic myocardium through patent channels; induction of angiogenesis; and regional denervation. We sought to determine whether transmyocardial laser revascularization modifies afferent and efferent axonal function within the affected myocardium. METHODS: Studies were performed in 9 dogs that were artificially ventilated and underwent thoracotomy. Changes in ventricular dynamics and intrinsic cardiac neuronal activity were monitored before and after creating 20 transmural channels in the left ventricular ventral free wall with a holmium:yttrium-aluminum-garnet laser in response to three stimuli: application of veratridine or bradykinin to the epicardial sensory neurites of intrinsic cardiac afferent neurons; sympathetic or parasympathetic efferent neuronal activation either electrically (4 V, 10 Hz, 5 ms) or chemically (nicotine, 5 microg/kg intravenously), and direct cardiomyocyte beta-adrenergic receptor stimulation (isoproterenol hydrochloride, 5 microg intravenously). RESULTS: Sensory neurites of right atrial afferent neurons in the studied epicardial region responded similarly to chemical stimulation before and after transmyocardial laser revascularization. Transmyocardial laser treatment did not reduce local ventricular contractile responses to direct activation of sympathetic or parasympathetic efferent neurons by electrical or chemical means, nor did it affect cardiomyocyte augmentor responses elicited by exogenous beta adrenergic receptor challenge. CONCLUSIONS: As transmyocardial laser revascularization does not affect afferent or efferent axonal function in the affected ventricle, the efficacy of this form of therapy cannot be ascribed to local denervation. PMID- 10475414 TI - Coronary revascularization without cardiopulmonary bypass in patients with pectus excavatum. AB - BACKGROUND: Coronary revascularization in patients with pectus excavatum is technically difficult through a median sternotomy secondary to the posterior displacement of the sternum and the asymmetric angulation that it produces. The role of minimally invasive coronary artery bypass grafting (MIDCABG) in this subset of patients was evaluated. METHODS: In 1998, four patients with pectus excavatum underwent revascularization of the left anterior descending artery without cardiopulmonary bypass through a left anterior minithoracotomy. RESULTS: All patients underwent the procedure without intraoperative complications and postoperative angiography demonstrated adequate graft patency. CONCLUSIONS: The advantages of MIDCABG in patients with pectus excavatum is the superior exposure to the LAD and LIMA and avoidance of a median sternotomy and cardiopulmonary bypass. This procedure is deemed safe and effective in patients with such deformities of the chest wall. PMID- 10475415 TI - Aprotinin inhibits thrombin formation and monocyte tissue factor in simulated cardiopulmonary bypass. AB - BACKGROUND: Aprotinin reduces perioperative bleeding after open heart surgery, primarily by inhibiting fibrinolysis. In addition, the drug has both procoagulant and anticoagulant effects that involve complex reactions of coagulation proteins and cells that are incompletely understood. This study tests the hypothesis that aprotinin has an anticoagulant effect on the extrinsic coagulation pathway. METHODS: Human heparinized blood was recirculated through a membrane oxygenator with and without high concentrations of aprotinin (18.4 microM). Serial plasma samples were obtained at intervals up to 240 minutes. RESULTS: Aprotinin significantly reduced the progressive increase in prothrombin fragments (F1.2) and thrombin-antithrombin complex beginning immediately. Aprotinin also significantly reduced monocyte expression of tissue factor and Mac-1. Aprotinin did not significantly reduce factor VII or factor VIIa. CONCLUSIONS: During simulated cardiopulmonary bypass, aprotinin immediately inhibits kallikrein and thrombin formation via the intrinsic coagulation pathway. Later, aprotinin inhibits monocyte expression of tissue factor and the extrinsic coagulation pathway. The ability of aprotinin to inhibit monocyte tissue factor provides a means to reduce thrombin formation in blood aspirated from the wound during open heart surgery. PMID- 10475416 TI - A sprayable hemostat containing fibrillar collagen, bovine thrombin, and autologous plasma. AB - BACKGROUND: A new sprayable hemostat, CoStasis hemostatic device (Cohesion Technologies Inc, Palo Alto, CA), consisting of collagen, thrombin, and autologous plasma was tested versus fibrin sealant and collagen hemostatic sponges. Performance was also monitored as fibrinogen and platelets were depleted from the plasma. In addition, the strength of the CoStasis gel, its sealing ability, its fibrillar structure, and its platelet-aggregating ability were investigated. METHODS: Hemostatic performance was determined with an in vivo bleeding rabbit kidney and spleen model. Differential scanning calorimetry and electron microscopy were used to analyze collagen structure. Sealing ability was determined with a burst-test apparatus. RESULTS: In the in vivo model, CoStasis was superior to fibrin sealant and collagen sponges in achieving a rapid time to hemostasis. The formulation continued to perform well when either platelets or fibrinogen was depleted. CoStasis formed weaker gels than fibrin sealant and could withstand only modest pressures. The collagen in the formulation had a fibrillar structure that was shown to aggregate human platelets. CONCLUSIONS: CoStasis, with the two platelet activators collagen and thrombin in addition to the thrombin-catalyzed formation of fibrin and the sealing properties of the soft gel, provides an excellent atraumatic hemostat. PMID- 10475417 TI - Tracheostomy in cardiosurgical patients: surgical tracheostomy versus ciaglia and fantoni methods. AB - BACKGROUND: Patients requiring prolonged mechanical ventilation are not uncommon in a cardiosurgical intensive care unit. Elective tracheostomy is considered the airway treatment of choice in these patients. METHODS: To evaluate different techniques for tracheostomy, we prospectively investigated 120 patients who had conventional open (n = 40), minimally invasive percutaneous dilatational (n = 40), or translaryngeal (n = 40) tracheostomy techniques. The main areas of investigation included oxygenation index (partial pressure of arterial oxygen divided by fraction of inspired oxygen), complications, infection, and cost. RESULTS: The oxygenation index decreased in almost every patient, regardless of the technique used, but the extent of decrease was significantly lower in both minimally invasive techniques compared with the conventional method. Overall complication rate was 12.5% both in open tracheostomy and in percutaneous dilatational tracheostomy, whereas no complications occurred in translaryngeal tracheostomy procedures. Bacterial contamination of the tracheostomy site was found in 35% of the open tracheostomies, whereas no infection was seen in percutaneous dilatational or translaryngeal tracheostomies. In terms of costs, PDT ($506) and TLT ($362) were both much cheaper than open tracheostomy ($699). CONCLUSIONS: Percutaneous dilatational and translaryngeal tracheostomies are safe and cost-effective procedures that can be done easily at the patient's bedside and thus are attractive alternatives to conventional surgical tracheostomy in long-term airway access in a cardiosurgical intensive care unit. PMID- 10475418 TI - On-pump versus off-pump coronary revascularization: evaluation of renal function. AB - BACKGROUND: Coronary revascularization with cardiopulmonary bypass has the potential risk of renal dysfunction related to the nonphysiologic nature of cardiopulmonary bypass. Recently, there has been a revival of interest in performing myocardial revascularization on the beating heart and we investigated whether this prevents renal compromise. METHODS: A prospective, randomized, controlled trial was performed in 50 patients (45 males, mean age 61+/-3.7 years) undergoing elective coronary artery bypass grafting. Patients were randomly assigned to conventional revascularization with cardiopulmonary bypass (on pump) or beating heart revascularization (off pump). Glomerular and tubular function were assessed up to 48 hours postoperatively. RESULTS: There were no deaths, myocardial infarctions or acute renal failure in either group. Glomerular filtration as assessed by creatinine clearance and the urinary microalbumin/creatinine ratio was significantly worse in the on pump group (p < 0.0004 and 0.0083, respectively). Renal tubular function was also impaired in the on pump group as assessed by increased N-acetyl glucosaminidase activity (p < 0.0272). CONCLUSIONS: These results suggest that off pump coronary revascularization offers a superior renal protection when compared with conventional coronary revascularization with cardiopulmonary bypass and cardioplegic arrest in first time coronary bypass patients. PMID- 10475419 TI - Effects of cyanosis and hypothermic circulatory arrest on lung function in neonatal lambs. AB - BACKGROUND: Lung function is often impaired after cardiac surgery and cardiopulmonary bypass (CPB), particularly in chronically cyanotic patients. This study aimed to evaluate lung function in a surgically created chronic cyanotic neonatal lamb model after CPB and deep hypothermic circulatory arrest (DHCA) and to assess the role of nitric oxide (NO) in the pathogenesis of increased pulmonary vascular resistance. METHODS: A chronic cyanosis model was surgically created in 7 lambs (4.7+/-0.8 days old) by anastomosing the pulmonary artery (PA) to the left atrium (LA). Another 7 lambs underwent a sham operation (control). One week later, the animals underwent shunt takedown and CPB with 90 minutes of DHCA at 18 degrees C. Cardiac index (CI), pulmonary vascular resistance index (PVRI), lung dynamic compliance (Cdyn), alveolar-arterial oxygen difference (AaDO2), left atrial plasma nitrate/nitrite (NO metabolites) levels, and pulmonary cGMP production (concentration difference between LA and PA) were measured before CPB and at 1 and 2 hours after reperfusion. RESULTS: The cyanosis model consistently produced significantly lower arterial oxygen tension (34.8+/ 2.3 vs 93.1+/-8.8 torr in control, p < 0.001) and Qp/Qs (0.6+/-0.1 vs 1.0+/-0.0 in control, p < 0.001) than controls. Postoperative PVRI was significantly lower in the cyanosis group than in controls, although CPB with DHCA significantly elevated PVR in both cyanotic and control animals. There were no significant differences in AaDO2 and Cdyn after CPB between groups. The level of NO metabolites did not change before or after CPB in either cyanotic or acyanotic animals. NO metabolite levels tended to be higher in the cyanotic animals (p = 0.08). There was no significant difference in pulmonary cGMP production between both groups. CONCLUSIONS: These findings suggest that CPB with DHCA, per se, does not affect NO production in cyanotic or acyanotic neonatal lambs but causes increased PVR in both groups. Chronic cyanosis does not result in reduced pulmonary function after CPB with DHCA, and is associated with lower PVR. The mechanism may involve an increased NO production in cyanotic animals. PMID- 10475420 TI - Pediatric coronary artery bypass for Kawasaki congenital, post arterial switch, and iatrogenic lesions. AB - BACKGROUND: Pediatric coronary artery bypass (PCAB) has been recently employed for expanding indications to treat acquired, congenital, post arterial switch, and other iatrogenic pediatric coronary artery problems. METHODS: Between 1987 and 1998, 3 infants and 13 children (n = 16, mean age 6.1 years, range 2 months 18 years) underwent one or more internal thoracic artery (ITA) to coronary artery (CA) bypass grafts for Kawasaki disease (n = 4), congenital lesions (n = 3), post arterial switch (n = 4), and other iatrogenic obstructions (n = 5). Proximal left main CA arterioplasty was performed concurrently with ITA-CA bypass in 4 patients. RESULTS: Survival is 93.8%. All bypass grafts in surviving patients are patent 2 months-11 years postoperation. The 11 elective patients are well (NYHA I II). The 5 emergent operations were performed in 2 infants and 3 adolescents who had poor ventricular function prior to ITA-CA bypass due to iatrogenic injuries in 3, congenital critical left main stenosis in 1, and intraoperative iatrogenic coronary injury in 1. The 3 adolescents fared worse, resulting in death in the first, cardiac transplantation in the second, and full recovery in the third. The 2 infants have steadily improving ventricular function. CONCLUSIONS: ITA-CA bypass can be successfully performed in infants and children for expanding elective and life-saving indications with excellent results. Poor preoperative ventricular function often persists, especially in those older children with iatrogenic injuries, and may result in death or cardiac transplantation. PMID- 10475421 TI - Infant cardiopulmonary bypass: a procoagulant state. AB - BACKGROUND: Procoagulant activity after cardiopulmonary bypass (CPB) in infants may predispose to thrombotic and bleeding complications. The induction of tissue factor and prothrombinase activity on endothelial cell membranes is a primary step in the activation of the extrinsic clotting cascade. The purpose of this study is to characterize the fibrinolytic and endothelial procoagulant state in infants undergoing congenital cardiac repairs with and without CPB. METHODS: Fourteen infants (aged 1 to 12 weeks) underwent repair of congenital cardiac defects. Two patients had closed procedures (controls) and 12 had open cardiac procedures. Serum samples were taken before and after CPB, 1, 4, and 24 hours after CPB. Tissue plasminogen activator, plasminogen activator inhibitor-1, interleukin-1beta, interleukin-6, plasma tissue factor, and factor V levels were measured. Human umbilical vein endothelial cell cultures were incubated with serum taken from the above time points and assayed for induction of tissue factor and prothrombinase activity. RESULTS: Control patients had no change from preoperative values in any of the parameters examined. In experimental patients, tissue plasminogen activator levels peaked at 1 hour after CPB and then decreased to normal by 24 hours. Plasminogen activator inhibitor-1 levels peaked at 4 hours after CPB and returned to baseline by 24 hours. The plasma of all patients had no intrinsic tissue factor activity. Induction of tissue factor activity on umbilical vein endothelial cells peaked immediately and again at 24 hours, whereas prothrombinase activity peaked early and stayed elevated. Serum factor V levels were significantly reduced after CPB, but returned to near baseline levels by 24 hours. CONCLUSIONS: Cardiopulmonary bypass is associated with derangement of the coagulation and fibrinolytic systems in infants. The serum of these patients promotes the induction of endothelial procoagulant activity, suggesting that there may be a hypercoagulable state in the postbypass period. PMID- 10475422 TI - Intermediate-term results in pediatric aortic valve replacement. AB - BACKGROUND: Aortic valve replacement (AVR) in children is now more commonly performed with human tissue valves. METHODS: The results of 100 consecutive pediatric AVRs (50 mechanical, 50 human) were reviewed. RESULTS: There were five perioperative deaths in the mechanical group and one in the human group (p = 0.2). Late complications in the mechanical group included 4 late deaths, 2 cases of endocarditis, 3 thromboembolic complications, and 10 reoperations on the aortic valve. In the human group, there were no late deaths, 2 reoperations for allograft aortic valve deterioration (both in Marfan's patients), and 1 reoperation for allograft pulmonary valve stenosis. Four-year actuarial survival was 83% in the mechanical group and 98% in the human group (p = 0.02). Four-year actuarial survival free of all valve-related complications was 61% in the mechanical group and 88% in the human group (p = 0.008). CONCLUSIONS: Human valves in children requiring AVR provide superior intermediate-term survival and freedom from valve-related complications compared to mechanical valves. Marfan's syndrome may represent a rare remaining contraindication for human AVR in children. PMID- 10475423 TI - Current results with pediatric heart transplantation. AB - BACKGROUND: Cardiac transplantation is an accepted treatment for children with end-stage heart failure or complex or inoperable congenital defects. METHODS: Since 1988, 95 transplants have been performed in 89 children aged 4 days to 18 years (median 6.9 years, 42 patients 0-5 years). Fifty-eight (61%) had congenital or acquired heart disease, 31 (33%) had idiopathic cardiomyopathy, and 6 (6%) were retransplants. Fifty-seven of the patients had prior cardiac surgery with a range of one to eight procedures (mean 3.4 procedures/patient). At the time of transplantation, 53 (56%) were United Network for Organ Sharing (UNOS) status I, including 23 children on mechanical ventilation and 4 with mechanical circulatory support. RESULTS: Thirty-day survival in this group was 96%. Posttransplant results showed a median time of ventilation of 1 day (mean 3.0+/-5.7 days), median duration of inotropic support of 2 days (mean 2.7+/-2.3 days), median intensive care unit (ICU) stay of 4 days (mean 6.9+/-9.6 days), and median hospitalization of 9 days (mean 14.3+/-13.9 days). Follow-up from 1 month to 10.3 years (mean 3.1 years) has demonstrated a 1-year actuarial survival of 79% and a 5-year actuarial survival of 69%. Rejection, both acute and chronic, accounted for the vast majority of deaths. CONCLUSIONS: Pediatric heart transplantation can be accomplished with excellent early survival despite multiple prior cardiac operations and relative severity of illness. Parameters such as postoperative ventilation, inotropic support, ICU stay, and hospitalization can be kept at reasonable levels with acceptable long-term results, although rejection remains a serious problem. PMID- 10475424 TI - Pediatric open heart operations without diagnostic cardiac catheterization. AB - BACKGROUND: Echocardiography allows for an adequate noninvasive preoperative evaluation in an increasing proportion of pediatric patients before open heart operations. The present study assessed the diagnostic accuracy of both invasive and noninvasive preoperative evaluation in children with congenital heart disease. METHODS: We prospectively evaluated the accuracy of preoperative noninvasive or invasive diagnostic methods. Preoperatively established diagnosis was compared with the intraoperative diagnosis made by surgical inspection and routine perioperative transesophageal echocardiography. RESULTS: During the study period of 30 months (ending in December 1997) 209 open-heart procedures were performed. Eighty-one patients (39%) were in the first year of life at the time of surgery, and 43% of all patients had symptoms. Noninvasive preoperative diagnosis using echocardiography was done exclusively in 142 patients (68%). Of the 67 children who had preoperative catheterization, 4 (6%) showed an additional intraoperative finding that modified the surgical approach in 2 of them. In the 142 patients who had echocardiographic preoperative assessment, the surgeons were confronted with a previously undiagnosed finding in 12 patients (8.5%). The finding was considered significant (prolongation of cardiopulmonary bypass time) in 2 patients and might have affected the outcome in 1 of them, a neonate with transposition of the great arteries and a preoperatively undiagnosed intramural coronary artery, who died postoperatively despite a technically adequate repair. CONCLUSIONS: In many infants and children, diagnostic work-up before open heart operations could be adequately based on an exclusively noninvasive basis by relying on echocardiography alone. PMID- 10475425 TI - The long-term results of commissure plication annuloplasty for congenital mitral insufficiency. AB - BACKGROUND: Mitral valve repair in the pediatric population remains demanding because of a diversity of apparatus anomalies and the young age of the patients. METHODS: We reviewed our clinical results for mitral valve repairs for congenital mitral insufficiency. Forty-nine consecutive patients aged 2 months to 34 years (mean, 4.4 years) had mitral valve repair between June 1984 and December 1996. Forty-one patients (83.7%) had associated cardiac anomalies. The predominant pathologies for the regurgitations were chordal anomalies in 34 patients (69%), annular dilatation in 8 (16%), and leaflet anomalies in 7 (14%). Mitral valve repair included commissure plication annuloplasty in 43 patients (88%), modified DeVega in 11, cleft closure in 5, plication of the anterior leaflet in 3, triangular resection of the anterior leaflet in 2, chordal shortening in 1, and placement of artificial chordae in 1. Several combined techniques were required in 19 patients. RESULTS: There were no early or late deaths. The follow-up period was from 6 to 166 months (mean, 88.4 months). Forty-seven patients (95.9%) were in New York Heart Association class I. The long-term echocardiographic studies showed that 2 of 30 patients without reoperation had moderate regurgitation. The actuarial freedom from reoperation was 85.6% (95% confidence limits, 72.8%, 98.4%) at 13 years. Five patients (10.2%) required valve replacement from 13 days to 75 months after the valve repair. Two patients had cerebral ischemic events as a result of cardiomegaly and atrial fibrillation. CONCLUSIONS: Valve repair for congenital mitral insufficiency gave adequate results in combination with commissure plication annuloplasty and other techniques with excellent long-term functional status. PMID- 10475426 TI - Valved homograft conduit repair of the right heart in early infancy. AB - BACKGROUND: Valved homograft conduit repair in neonates and young infants creates a physiologically normal biventricular circulation, and unlike shunts, avoids surgery on the branch pulmonary. METHODS: Retrospective chart review was used for 84 patients operated on between 1990 and 1995 (mean age 26+/-28 days, mean weight 3.3+/-0.8 kg) undergoing homograft conduit repair in the first 3 months of life. Cases were divided into simple and complex, eg, absent pulmonary valve syndrome or associated interrupted arch. Mean homograft size was 9.0+/-2 mm. RESULTS: Early mortality was 4.7% (simple) and 30% (complex). Mean hospital stay was 18 days. Mean follow-up was 34 months. Thirty-seven (47%) patients underwent conduit replacement. Median time to reoperation was 3.1 years. Mean size of replacement homograft was 17+/-2 mm. There were no deaths at reoperation. Mean hospital stay at conduit change was 6.3 days. Probability of survival at 5 years is 75%. CONCLUSIONS: Biventricular repair employing a conduit can be performed safely in noncomplex anomalies in the first 3 months of life. Time interval until repeat surgery is relatively short but equal or greater than that with most palliative procedures. PMID- 10475427 TI - Left ventricular growth in selected hypoplastic left ventricles: outcome after repair of coarctation of aorta. AB - BACKGROUND: Models that predict survival in neonates with left ventricular hypoplasia and critical aortic stenosis may not be applicable to neonates with left ventricular hypoplasia and coarctation. METHODS AND RESULTS: We report 8 infants with severe aortic coarctation and left ventricular hypoplasia. Mean age was 18 days (range 1-48 days), and mean weight was 3.5 kg (range 2.7-4.3 kg). Associated diagnoses included mild aortic stenosis (4), ventricular septal defect (2), and venous anomalies (2). All had coarctation repair as a primary procedure (3 of these had concomitant intracardiac procedures); 7 had subsequent operations. All are alive and well 1.1-6.7 years (mean 3.1 years) after the first surgery. Progressive increases were observed in aortic and mitral diameters, and in left ventricular dimensions, areas, and volumes when the preoperative, earliest postoperative, and most recent echocardiograms were compared. CONCLUSIONS: Despite severe left ventricular hypoplasia, a two-ventricle repair is possible in selected cases. The prognostic criteria for left ventricular hypoplasia in critical aortic stenosis may not be applicable to infant coarctation. Relief of coarctation may result in the growth of the very small left ventricle, especially when the aortic root and mitral diameters are satisfactory. PMID- 10475428 TI - Prosthetic valve replacement in Ebstein's malformation: 30-year follow-up. AB - This report addresses a 42-year-old asymptomatic patient who underwent prosthetic replacement of his tricuspid valve 31 years previously for Ebstein's malformation. A ball valve prosthesis was implanted to replace the abnormal valve. Furthermore, he had concomitant closure of a large atrial septal defect with a perforated patch together with plication of the atrialized ventricular segment. The patient is asymptomatic, and it remains noteworthy that he has not been on coumadin for about 30 years. PMID- 10475429 TI - AP window and anomalous origin of right coronary artery from the window. AB - Aortopulmonary window (APW) is a rare malformation. We recently operated on a child with APW, ventricular septal defect, right aortic arch, and anomalous right coronary artery from the APW. This patient also had a chromosomal abnormality. He underwent the repair of this complex lesion in a staged operation. PMID- 10475430 TI - Norwood procedure without circulatory arrest. AB - In the Norwood procedure for hypoplastic left heart yndrome, the distal descending thoracic aorta was cannulated just superior to the diaphragm through median sternotomy. In combination with cerebral perfusion through the graft anastomosed to the innominate artery, which was used as a systemic-to-pulmonary shunt later, this technique enabled us to completely avoid circulatory arrest and deep hypothermia throughout the operation. PMID- 10475431 TI - Gastric tube-to-tracheal fistula closed with a latissimus dorsi myocutaneous flap. AB - A gastric tube-to-airway fistula is a very rare complication after esophageal reconstruction. A patient with a gastric tube-to-tracheal fistula that developed more than 9 years after surgery for cancer of the cervical esophagus was treated with transposition of a pedicled latissimus dorsi myocutaneous flap. Careful perioperative respiratory management helped save the patient's life. PMID- 10475432 TI - Right ventricular thrombosis early after bidirectional Glenn shunt. AB - Thrombosis in the right ventricle occurred early after bidirectional superior cavopulmonary shunt in 2 patients with pulmonary atresia with intact ventricular septum and major right ventricular coronary artery communication, and perioperative brain infarction occurred in 1 patient. Clinicians should be aware of the hazards of this potentially lethal complication, and transfusion of platelets and fresh plasma should be minimized. Although the hemodynamic state is good, echocardiography should be performed frequently and strict anticoagulation should be started as early as possible. PMID- 10475433 TI - A malignant primary cardiac pheochromocytoma. AB - We report the case of a malignant primary cardiac pheochromocytoma treated by adjuvant cytotoxic chemotherapy after surgical resection, with a 5-year survival. There is no specific chemotherapy for malignant pheochromocytoma, but because it has the same embryologic origin as neuroblastoma, we used similar chemotherapy. Because of unexpected malignancy potential, we think that total and meticulous resection of the tumor must be done. PMID- 10475434 TI - Tension pneumocephalus after excision of posterior mediastinal mass. AB - Although paraplegia after a posterolateral thoracotomy has been described as a serious complication, the occurrence of tension pneumocephalus after excision of posterior mediastinal mass has also been reported as another life-threatening complication. We report one such case. PMID- 10475436 TI - Combined Rastelli and atrial switch procedure: a 10-year follow-up. AB - We performed a combined atrial (Mustard) and ventricular (Rastelli) repair on a previously palliated patient with situs inversus, atrioventricular discordance, ventricular septal defect, and pulmonary atresia. The suitability and durability of this operative strategy is supported by the satisfactory hemodynamic and functional status of the patient 10 years later. PMID- 10475435 TI - Epidural abscess: a delayed complication of esophageal stenting for benign stricture. AB - A case of anterior cervical epidural abscess associated with perforation of an endoscopically placed esophageal stent is presented. Although delayed esophageal perforation is a known complication of endoscopic stenting, no cases presenting with epidural abscess have yet been reported. The increasing application of endoscopic stenting for benign esophageal strictures provides greater opportunity for this type of delayed complication. PMID- 10475437 TI - Rapid two-stage repair of S,L,L, ventricular septal defect, pulmonary atresia, Ebstein anomaly of the tricuspid valve, and stenotic pulmonary arteries. AB - The repair of a corrected transposition of the great arteries, ventricular septal defect, and pulmonary atresia is presented. An Ebstein anomaly of the tricuspid valve, dextrocardia, and severe distortion of the pulmonary arteries complicated the surgical procedure, which was performed in two stages. Reconstruction of the pulmonary arteries and a bidirectional cavopulmonary anastomosis were performed first; Rastelli and hemi-Mustard procedures completed the correction. The rationale and the possible indications of this "one and a half ventricle" repair are discussed. PMID- 10475438 TI - Separate revascularization of the visceral arteries in thoracoabdominal aneurysm repair. AB - We successfully repaired a ruptured aortic patch containing the visceral arteries after thoracoabdominal aneurysm repair in a young patient with Marfan syndrome. This unexpected and tricky situation was treated by separate revascularization of each visceral artery using small-caliber prosthetic grafts as interposition between the aortic prosthesis and the ostia of the visceral arteries. PMID- 10475439 TI - An unusual iatrogenic cause of right coronary air embolism. AB - A 62-year-old woman undergoing redo mitral valve replacement was noted to have persistent intracardiac air following standard deairing procedures. Transesophageal echocardiography (TEE) identified air bubbles entering the left atrium from the right superior pulmonary vein. Exploration of the pleural cavity revealed a fistula between the pulmonary parenchyma and the right superior pulmonary vein caused by the atriotomy closure suture transfixing the edge of the lung, which was repaired with immediate disappearance of the air emboli. This demonstrates that transesophageal echocardiography is an invaluable aid to ensuring complete deairing after open heart procedures. PMID- 10475440 TI - Heparin-induced thrombocytopenia in coronary bypass surgery. AB - We report on a 51-year-old man with severe two-vessel coronary disease and an ejection fraction of 15% who presented with myocardial ischemia and heparin induced thrombocytopenia after coronary angioplasty. Before coronary bypass surgery, the antithrombin agent argatroban was used for anticoagulation and an intraaortic balloon pump was inserted. Direct coronary bypass surgery was performed to the left anterior descending artery and to the posterior descending artery using the 'Octopus' tissue stabilization device (Manfrotto, Feltre, Italy). The postoperative course was uneventful and associated with normal platelet counts. The patient was discharged on the 6th postoperative day. PMID- 10475441 TI - Immune-deficient bronchiectasis associated with X-linked lymphoproliferative disease. AB - Bronchiectasis may occur with various congenital and acquired immunodeficiency diseases. The association of bronchiectasis and the X-linked lymphoproliferative disease (XLP), also known as Duncan's disease is unknown. We describe the case of a 39-year-old man with XLP, the oldest surviving, who developed chronic bronchiectasis with hemoptysis and required a pneumonectomy to control his symptoms. PMID- 10475442 TI - Left ventricular fibroma masquerading as postinfarction myocardial rupture. AB - A large left ventricular fibroma was encountered perioperatively for what was presumed to be a sealed ventricular rupture after thrombolytic therapy for an acute myocardial infarction. We review the pertinent literature concerning the diagnosis of ventricular rupture and this rare benign tumor of the heart. PMID- 10475443 TI - Transcatheter closure of a patent foramen ovale following mitral valve replacement. AB - We report the successful closure of a postoperative patent foramen ovale in a patient who underwent coronary artery bypass grafting and mitral valve replacement for severe mitral insufficiency. The postoperative course was complicated by severe hypoxemia due to a large patent foramen ovale. The patient underwent transcatheter closure with the Das Angel Wings transcatheter occluder (Microvena Corporation, White Bear Lake, MN) with immediate improvement. PMID- 10475444 TI - Images in cardiothoracic surgery. Herniation of emphysematous bulla through a chest tube site. PMID- 10475445 TI - Vascular clamp for hemostasis and stabilization during minimally invasive direct coronary artery bypass. AB - A soft vascular clamp was used for hemostasis and stabilization of the operative field during minimally invasive direct coronary artery bypass (MIDCAB). The instrument was gently applied so that it clamps the coronary artery by grasping the adjacent myocardium. The method offered dry and stable operative field without a special instrument or technique. The surgical results have been satisfactory. We found application of the vascular clamp to be very helpful for MIDCAB. PMID- 10475446 TI - Emergency cannulation for proximal perfusion in descending thoracic aorta procedures. AB - Acute cardiac failure during descending thoracic aorta operations, although rare, may have catastrophic consequences. Under these circumstances, the use of partial veno arterial bypass is advantageous, allowing an assisted perfusion of both proximal and distal circulation districts. Traditionally, the ascending aorta or the aortic arch are the preferred sites of cannulation for proximal arterial reinfusion, but some limitations, such as extensive calcifications or extreme fragility of these segments, may hamper or at least delay this action. Herein, we describe a simple technique for rapid cannulation of proximal aorta in emergency circumstances. PMID- 10475447 TI - Transmanubrial approach with antero-lateral thoracotomy for apical chest tumor. AB - Transmanubrial osteomuscular sparing approach (TMA) has been recently proposed for the treatment of apical chest tumor to allow a safer subclavian artery control with a less invasive procedure for the patient. The present technique combines the antero-lateral muscle-sparing thoracotomy with TMA for lung cancer patients in whom extended resection of cervico-thoracic structures as well as anatomic lung resection and radical lymph nodes dissection are required. PMID- 10475448 TI - Coronary artery disease in identical twins. AB - The incidence of coronary artery disease in identical twins is unknown. The knowledge of ischemic heart disease in this patient population is represented by case reports. There is considerable controversy regarding the environmental and genetic factors that play a role in the pathogenesis of this disorder. The natural history of coronary artery disease in an asymptomatic twin with a symptomatic counterpart is unclear. We present a case report and literature review of coronary artery disease in identical twins. On the basis of our patient and the information of other patients, asymptomatic twins of symptomatic counterparts require aggressive assessment and management for occult coronary artery disease. PMID- 10475449 TI - The Society of Thoracic Surgeons National Congenital Heart Surgery Database Report: analysis of the first harvest (1994-1997). AB - This analysis summarizes the first report of the Society of Thoracic Surgeons National Congenital Heart Surgery Database Committee in association with Summit Medical Systems. Twenty-four centers joined the program at various dates of entry resulting in 18,894 enrolled patient records. This report compiled the relevant clinical features of 18 congenital heart categories over a 4-year period (1994 1997), which included 8,149 patient records. The data analyses are largely descriptive in character. Missing data points were described and not omitted in the analysis. Statistical analysis was not performed due to missing data points in some categories. Certain trends, however, could be identified and are discussed. The first Society of Thoracic Surgeons National Congenital Heart Surgery Database Report has succeeded in establishing a finite record that can be improved to establish universal national and international utility, risk stratification, and scholarly outcome analyses. PMID- 10475450 TI - Gender and coronary artery bypass mortality. PMID- 10475451 TI - Iatrogenic mammary arteriovenous fistula caused by a sternal wire. PMID- 10475452 TI - Bilateral internal thoracic arteries and diabetes. PMID- 10475453 TI - To use or not to use propranolol for prophylaxis of postthoracotomy arrhythmias. PMID- 10475454 TI - Video-assisted minimal access in excision of left atrial myxoma. PMID- 10475455 TI - Minding the chaperones. PMID- 10475456 TI - Asymptomatic traumatic aortic rupture. PMID- 10475457 TI - Troponin I and autotransfusion of shed mediastinal blood in redo cardiac operation. PMID- 10475458 TI - Coronary endarterectomy on beating heart. PMID- 10475459 TI - Avoiding injury to a patent left internal thoracic artery during reoperative coronary artery bypass grafting. PMID- 10475460 TI - Inferior epigastric artery. PMID- 10475461 TI - A miniature implantable axial flow ventricular assist device. AB - BACKGROUND: Since 1984, in collaboration with NASA engineers, we developed an axial flow pump that is 86 mm long, 22 mm wide, weighs 95 g, produces a flow of 5 to 6 L/min against a 100-mm Hg pressure at about 10,000 rpm, and requires less than 10 W of power. METHODS: The pump has been implanted in 9 calves with the inlet cannula inserted into the left ventricle and the outlet cannula, consisting of an albumin-coated Dacron graft, attached by end-to-side anastomosis to the descending thoracic aorta. RESULTS: All animals showed normal behavior until they were killed 1 to 3 months after operation. At autopsy, systemic studies of vital organs demonstrated no evidence of thromboembolism; the rpm of the pump was maintained between 9,000 and 10,000; the wattage ranged between 7 and 9; the output between 4 and 5 L/min; the hemoglobin was maintained between 32 and 35 mg/dL; the plasma-free hemoglobin ranged between 0.5 and 3 mg/dL; the BUN ranged between 8 and 14 mg/dL; the creatinine remained less than 1 mg/dL; and bilirubin studies were within normal limits. Bearing wear-tear tests up to about 5 months have been negative. CONCLUSIONS: The performance characteristics of the pump implanted in calves up to 90 days are highly gratifying, particularly in terms of pump output of 5 L/min, an index of hemolysis well within normal limits, and absence of thromboembolism. PMID- 10475462 TI - Mechanical circulatory support systems: past, present, and future. PMID- 10475463 TI - Ventricular assist device programs: design and function. AB - Ventricular assist device programs can be structured in a variety of models and can make use of personnel derived from a broad spectrum of clinical and hospital based practice. When aggressively and proactively negotiated with payers, these programs can be cost-effective and thoroughly self-sustaining. PMID- 10475464 TI - The logistics and cost-effectiveness of circulatory support: advantages of the ABIOMED BVS 5000. AB - BACKGROUND: In 1994, the ABIOMED BVS 5000 was incorporated into our acute cardiac assist armamentarium. This report is a general overview of our experience. A hypothetical cost analysis focusing on specific devices and device-related personnel contrasted the BVS 5000 with our prior model of centrifugal pump use. METHODS: In 3 years, 22 patients were supported with the BVS 5000, as a biventricular assist device in 40%, right ventricular assist device in 27%, and left ventricular assist device in 32%. Indications were postcardiotomy support in 12, acute myocarditis in 2, bridge to transplant in 4, and failed heart transplant in 4. The cost analysis was performed retrospectively. The actual cost of disposable blood pumps, including replacement pumps, and cannulae constituted the BVS cost. The hypothetical centrifugal costs included the disposables, replacement cones, as well as the labor costs of the continuous perfusionist coverage. RESULTS: Of the 22 patients, 10 (45%) were weaned and 13 (59%) were successfully discharged. Five patients were transplanted while on BVS 5000 support, accounting for a higher rate of discharge. Comparison of "actual" BVS costs with "projected" centrifugal costs revealed differences based upon the intended application of the BVS. In bridge-to-transplant patients with long duration of support, the daily cost of support was dramatically lower with the BVS 5000. For short-term postcardiotomy support, acute myocarditis, or failed transplant, the differences were small. CONCLUSIONS: Because the BVS 5000 was readily managed by the intensive care unit nursing staff, this system displaced centrifugal systems in our program. Outcome measures of weaning and successful discharge were improved relative to our prior experience with centrifugal pumps. Even without taking indirect costs into account, the hypothetical cost analysis supported continued use of the BVS system for acute cardiac assistance. PMID- 10475465 TI - Acute and temporary ventricular support with BioMedicus centrifugal pump. AB - BACKGROUND: Short-term ventricular and pulmonary support can be provided by the Medtronic BioMedicus (Eden Prairie, MN) centrifugal pump, which is available in most cardiovascular surgery centers. This versatile pump can provide support during cardiopulmonary resuscitation, cardiopulmonary bypass, extracorporeal membrane oxygenation, and ventricular assistance. A common use of the pump is to provide ventricular assistance for patients after cardiotomy or cardiogenic shock. METHODS: From January 1986 to September 1995, 141 patients at The Methodist Hospital in Houston, Texas were placed on the BioMedicus centrifugal pump after postcardiotomy cardiac failure. Patient treatment and postimplant complications are discussed. RESULTS: Fifty-four percent of the patients were weaned; however, only 22% survived to discharge. There was a very high mortality rate in the early stage after support was discontinued, after weaning, and after device removal. CONCLUSIONS: A high incidence of complications and death is likely related to the period of attempted weaning from cardiopulmonary bypass before the initiation of ventricular support. When weaning a patient from the pump during cardiopulmonary bypass or during ventricular assistance, it is important to optimize preload, after-load, ventricular function, and cardiac rhythm. In patients who have had postcardiotomy support, avoiding fluid overload, low colloid oncotic pressure, hypoperfusion, and use of excessive inotropic and vasoactive medications improve results. PMID- 10475466 TI - Extracorporeal membrane oxygenation for adult cardiac support: the Allegheny experience. AB - BACKGROUND: A mix of cardiac assist options is necessary to meet the diverse indications for cardiac support in a comprehensive heart failure program. At our institution, an adult extracorporeal membrane oxygenation (ECMO) system comprising a centrifugal pump and hollow fiber membrane oxygenator is used for short-term and temporary cardiac assist. METHODS: Between December 1991 and August 1997, 82 adult cardiac patients were supported on ECMO. Indications for cardiac assist included postcardiotomy cardiogenic shock (PCCS, 55 patients), high-risk cardiology intervention (27 patients), perioperative cardiac graft failure (4 patients), and emergency cardiac resuscitation (6 patients). Data for analysis were collected by prospective completion of standardized ECMO report forms and retrospective review of hospital charts. RESULTS: The ECMO system was inexpensive to operate, uncomplicated to implant, and adaptable for diverse indications. Survival in PCCS was 20 of 55 patients (36%), with an increased survival rate of 56% (18 of 32 patients) in patients with PCCS after isolated coronary bypass. Catheter-based revascularizations were successfully performed in 26 of 27 (96%) high-acuity patients temporarily supported by ECMO, and 23 of 27 patients (85%) survived to discharge. Survival in the cardiac graft failure group was 2 of 4 (50%). No patient supported on ECMO for cardiac resuscitation survived. CONCLUSIONS: ECMO provides good cardiopulmonary and end-organ support; survival rates are similar to or higher than those seen with centrifugal pump support in comparable patient populations. PMID- 10475467 TI - Current strategy of temporary circulatory support for severe cardiac failure after operation. AB - BACKGROUND: After open heart operations about 1% of patients still need temporary circulatory support for severe cardiac failure, and over half of those patients die during or after the support. This study assessed the efficacy of the current strategy of circulatory support. METHODS: We assessed clinical outcome of 64 consecutive patients (1.5% of pump cases) who had temporary circulatory support associated with perioperative variables. The results were analyzed by logistic regression analysis. RESULTS: The weaning and discharge rates were 50.0% and 26.7% with venoarterial bypass, 76.2% and 57.1% with biventricular bypass, 87.5% and 37.5% with isolated left ventricular bypass, and 60.0% and 40.0% with pulsatile left ventricular assistance, respectively. Logistic regression analysis identified presupport cardiogenic shock (odds ratio, 9.922) and support type (14.684) as factors significantly associated with nonweaning from the temporary support, and cardiogenic shock (28.268), support duration (2.948), and support type (14.184) as factors significantly associated with mortality during or after the circulatory support. CONCLUSIONS: The current strategy of temporary circulatory support improved clinical outcome of patients with severe cardiac failure. Early application of circulatory support before profound cardiogenic shock and proper selection of the support type might be key factors for successful circulatory support postoperatively after operation. PMID- 10475468 TI - Centrifugal pumps: description of devices and surgical techniques. AB - BACKGROUND: Because of simplicity of application, universal access, and low cost, centrifugal pumps are commonly used for short-term mechanical cardiac assist. Indications and techniques for application of this technology continue to evolve. METHODS: The clinical experience with 151 patients undergoing centrifugal mechanical cardiac assist at the University of Missouri-Columbia has been reviewed. We have compared commonly available centrifugal pumping systems in vitro and in vivo for characteristics that might distinguish them. RESULTS: Centrifugal pumps have been found to be well suited for use in surgery on the thoracic aorta, for extracorporeal membrane oxygenation and for postcardiotomy cardiac mechanical assist. Complications associated with centrifugal mechanical assist are predictable and common but potentially can be reduced by improved surgical techniques and anticoagulation strategies. In vitro and in vivo experimentation with available centrifugal pumps reveals nuances characteristic of each of the devices. CONCLUSIONS: All centrifugal pumps presently available are less destructive to blood cellular elements compared with roller pumps. With familiarity, all can function satisfactorily for short-term mechanical assist with no compelling evidence that favors any particular centrifugal pump system clinically available. Centrifugal pumps are ideally suited for left heart bypass during surgery on a thoracic aorta and for short-term application as may be required for postcardiotomy mechanical assist. Centrifugal pump technology should be part of the armamentarium of all cardiothoracic surgeons. PMID- 10475469 TI - Cardiopulmonary support and extracorporeal membrane oxygenation for cardiac assist. AB - BACKGROUND: Use of cardiopulmonary bypass for emergency resuscitation is not new. In fact, John Gibbon proposed this concept for the treatment of severe pulmonary embolism in 1937. Significant progress has been made since, and two main concepts for cardiac assist based on cardiopulmonary bypass have emerged: cardiopulmonary support (CPS) and extracorporeal membrane oxygenation (ECMO). The objective of this review is to summarize the state of the art in these two technologies. METHODS: Configuration of CPS is now fairly standard. A mobile cart with relatively large wheels allowing for easy transportation carries a centrifugal pump, a back-up battery with a charger, an oxygen cylinder, and a small heating system. Percutaneous cannulation, pump-driven venous return, rapid availability, and transportability are the main characteristics of a CPS system. Cardiocirculatory arrest is a major predictor of mortality despite the use of CPS. In contrast, CPS appears to be a powerful tool for patients in cardiogenic shock before cardiocirculatory arrest, requiring some type of therapeutic procedures, especially repair of anatomically correctable problems or bridging to other mechanical circulatory support systems such as ventricular assist devices. CPS is in general not suitable for long-term applications because of the small bore cannulas, resulting in significant pressure gradients and eventually hemolysis. RESULTS: In contrast, ECMO can be designed for longer-term circulatory support. This requires large-bore cannulas and specifically designed oxygenators. The latter are either plasma leakage resistent (true membranes) or relatively thrombo-resistant (heparin coated). Both technologies require oxygenator changeovers although the main reason for this is different (clotting for the former, plasma leakage for the latter). Likewise, the tubing within a roller pump has to be displaced and centrifugal pump heads have to be replaced over time. ECMO is certainly the first choice for a circulatory support system in the neonatal and pediatric age groups, where the other assist systems are too bulky. ECMO is also indicated for patients improving on CPS. Septic conditions are, in general, considered as contraindications for ECMO. CONCLUSIONS: Ease of availability and moderate cost of cardiopulmonary bypass-based cardiac support technologies have to be balanced against the significant immobilization of human resources, which is required to make them successful. PMID- 10475470 TI - Temporary pulsatile ventricular assist devices and biventricular assist devices. AB - BACKGROUND: During the past years several systems for mechanical circulatory support have become available. In this study we describe our experience with short-term and mid-term application of the ABIOMED and Thoratec device. METHODS: Since 1990 the ABIOMED BVS and since 1992 the Thoratec VAD have been applied to 75 and 103 patients, respectively, with postcardiotomy heart failure, as a bridge to-transplant procedure, and with different other indications. RESULTS: In the ABIOMED collective 25 of 50 patients (50%) with postcardiotomy heart failure and 1 of 4 patients with miscellaneous other indications could be discharged from hospital, 7 of 14 bridge-to-transplant patients (50%) underwent transplantation with a posttransplant survival of 86%. In the Thoratec collective 6 of 10 patients (60%) with postcardiotomy heart failure and 4 of 8 patients (50%) with miscellaneous indications could be discharged from hospital, 48 bridge-to transplant patients (74%) underwent transplantation with a posttransplant survival of 90%. CONCLUSIONS: The results show the versatility of the Thoratec VAD for short-term and mid-term application in patients with postcardiotomy heart failure and as a bridge-to-transplant procedure. The use of the ABIOMED device is not indicated for bridging patients to transplantation. Although in case of postcardiotomy heart failure, Thoratec is also superior to ABIOMED, the high costs of the Thoratec VAD limits its wide acceptance in this patient cohort. PMID- 10475471 TI - Bridge to transplantation: the Penn State experience. AB - BACKGROUND: During the past decade, ventricular assist devices as a bridge to transplantation have moved from the experimental arena to accepted therapy. Our institution has been at the forefront of the development of this technology and consequently has had extensive experience with the devices that are currently approved by the Food and Drug Administration for use as a bridge to heart transplantation. METHODS: The successful management of patients with assist devices hinges on patient and device selection as well as perioperative management strategies. The routine use of agents such as aprotinin, vasopressin, milrinone, and inhaled nitric oxide has contributed to successful management of these patients. We present our perspectives on the advantages and disadvantages of the Thermo-Cardiosystems HeartMate 1000 IP device and the Thoratec (Pierce Donachy) system. We also discuss our protocols and methods for patient selection, preoperative preparation, intraoperative strategy, and postoperative management that have resulted in improved patient outcomes. RESULTS: More than 60 device implantation procedures have been performed since the inception of our bridge to transplantation program. During this time, two thirds of our patients were successfully bridged to transplantation. Of these patients, 92% were alive at 1 month after transplantation, and 83% were alive at 1 year after transplantation. CONCLUSIONS: Both support systems are effective in supporting patients to heart transplantation. We have developed a preference for the Thermo-Cardiosystems HeartMate 1000 IP device because of its portability and associated better quality of life. However, the Thoratec device is the more versatile device, and circumstances exist when its use is clearly advantageous. In our institutional experience, outcome for bridging to transplantation has not been device dependent. PMID- 10475472 TI - 100 long-term implantable left ventricular assist devices: the Columbia Presbyterian interim experience. AB - BACKGROUND: The use of left ventricular assist devices (LVADs) as bridge to transplantation is now accepted as a standard of care for a subset of end-stage heart failure patients. Our interim experience with both pneumatically and electrically powered ThermoCardiosystems LVADs is presented to outline the benefits and limitations of device support as well as discuss its potential role as bridge to recovery and as destination therapy. METHODS AND RESULTS: Detailed records were kept prospectively for all patients undergoing LVAD insertion. One hundred LVADs were inserted over 7 years into 95 patients, with an overall survival rate of 75% and a transplantation rate of 70%. Four patients underwent device explant for recovered myocardial function. Three patients received LVADs as destination therapy in the ongoing REMATCH (Randomized Evaluation of Mechanical Assist Treatment for Congestive Heart failure) trial. Overall mean patient age was 51 years, and mean duration of support was 108 days. There were 25 device-related infections including the drive line, device pocket, and blood contacting surfaces. Cerebral vascular accidents and other embolic events occurred in 7 patients with six deaths. There were four device malfunctions and nine graft-related hemorrhages, resulting in six reoperations and three deaths. CONCLUSIONS: The use of long-term implantable LVADs will likely not be limited to bridge to transplantation. The REMATCH trial has commenced to study the role LVADs may have as an alternative to medical management. Furthermore, as the issues of myocardial recovery are examined, the "bridge to recovery" may be an important additional role for these assist devices. PMID- 10475473 TI - Bridge to transplant with the Novacor left ventricular assist system. AB - The Novacor left ventricular assist system is an implantable, electrically powered, wearable system that has been successfully used as a bridge to transplant since 1984. The pump is placed anterior to the left posterior rectus sheath with the percutaneous drive line positioned in the right lower abdominal wall. The seamless polyurethane blood sac is compressed by two solenoid-driven pusher plates. The inflow and outflow valves are porcine prostheses. The use of wearable external power packs facilitate patient mobility and rehabilitation before transplantation and may provide long-term support to selected patients as an alternative to transplantation. PMID- 10475474 TI - Implantation technique for the CardioWest total artificial heart. AB - The CardioWest total artificial heart is a pneumatically driven device that totally replaces the failing ventricles. It is currently undergoing clinical investigation as a bridge to heart transplantation in several centers throughout the world. A bilateral ventriculectomy is performed and the device is implanted. Blood flows are usually maintained at 6-8 L/min. Approximately 130 patients have undergone bridge to transplant with this device. Patient selection and excellent surgical technique are required for a successful outcome. A detailed description of the implantation technique is presented to facilitate the use of this technology. PMID- 10475475 TI - Preventing, minimizing, and managing postoperative bleeding. AB - BACKGROUND: Most bleeding in cardiovascular surgery is biological, not surgical, the result of disseminated intravascular coagulation in its latter phases. Disseminated intravascular coagulation bleeding affects all levels of the coagulation systems, requiring stabilization of the platelet system, balancing the procoagulant system and its regulation, and stopping the fibrinolytic system's activation. METHODS: The article discusses the multisystem protocol put into place at the Cardiovascular Surgery Department of La Pitie Hospital in Paris to diagnose and treat DIC bleeding so as to slow down causes of its occurrence and prevent further deterioration of the hemostatic systems. RESULTS: All biological bleeding was controlled, permitting detection of other causes of bleeding and allowing rational use of blood products. No thromboembolic accident occurred. There was an absence of iatrogenic bleeding. The protocol also detects disseminated intravascular coagulation in patients who did not bleed to take early and frequently lifesaving measures. CONCLUSIONS: This demonstrates the need to treat bleeding through a multisystem approach, monitoring its evolution by means of biological tests to be able to provide appropriate treatment. PMID- 10475476 TI - Infection during circulatory support with ventricular assist devices. AB - BACKGROUND: This study is a retrospective analysis of infections in patients supported by ventricular assist devices (VADs) as a bridge to cardiac transplantation. METHODS: Infections were assigned to four classes. Class I were patient-related nonblood infections, class II were blood-borne infections, class III were percutaneous site infections, and class IV were infections of intracorporeal VAD components. RESULTS: The cumulative number and incidence of infections were higher during the early VAD experience than in more recent patients (p < 0.05), although the cumulative number and incidence of class II and IV infections were similar in earlier and later patients. There were 28 serious (ie, class II and IV) infections in 9 patients who died, and 35 serious infections in 11 patients who survived until transplantation. Three of 4 patients with class IV infections died. A larger cumulative number of infections (ie, total class I-IV) was associated with more fungal isolates (p < 0.001) and more class II and IV infections (p < 0.02). Positive fungal cultures were obtained in 16 patients, but there were only 3 class III and 1 class IV fungal isolates. CONCLUSIONS: Infection remains an important problem for patients with VADs. Bloodstream infections (class II) can often be controlled by appropriate therapy. However, intracorporeal device infections (class IV) are associated with substantial morbidity and mortality. Optimal implant techniques together with optimal wound care, appropriate use of prophylactic antibiotics, and avoidance of infection in indwelling catheters remain the most practical means for minimizing the risk of VAD infection. PMID- 10475477 TI - Pseudointima in inflow conduits of left ventricular assist devices. AB - BACKGROUND: Explant analysis of left ventricular assist systems (LVAS) should permit a better evaluation of long-term evolution of materials and tissue healing in patients supported by mechanical devices and a precise understanding of embolic phenomena, observed clinically. METHODS: Five Novacor LVAS and their conduits have been explanted after 156 days (range 61-226 days) of mechanical support. The pseudo-intima (PI) developed in the inflow and outflow conduits was characterized microscopically, using monoclonal antibodies. RESULTS: The morphological aspects of PI were quite different in the inflow and outflow conduits. Blood coagulation between the basal surface of the PI and the Dacron tube, irregular collagen type I matrix with plasma infiltration, macrophages, and neutrophil granulocyte elastase characterized the nonadherent, loose, and potentially thrombogenic PI growth in the inflow conduit. The PI from collagen types I and IV with circumferentially oriented alpha-smooth muscle cell actin positive cells was anchored to the outflow conduits. CONCLUSIONS: The observations, which have to be confirmed by a more extensive study on a larger number of specimens, suggest the role of the biomaterial itself, as well as the configuration, physical characteristics, and rheology in the conduit. They also suggest that thromboembolic complications of LVAS may eventually be related to this host tissue response. PMID- 10475478 TI - Bridging from acute to chronic devices. AB - Mechanical circulatory support used as a bridge to chronic transplantation is now a firmly established treatment modality. Trials evaluating circulatory assist for permanent circulatory assist device implications are underway. A population of patients is beginning to emerge who are resuscitated using readily available "acute" devices and need long-term supporting "chronic" device to bridge either cardiac transplantation or permanent device implantation. Careful management of this new acute-to-chronic bridge group will maximize the chances of a successful long-term outcome. The best strategies for management may only unfold by later analysis of larger populations. This article summarizes the current utilization of acute and chronic devices, as well as the small but growing population of acute-to-chronic bridge patients, and suggests what seem today to be sound management principles for them. PMID- 10475479 TI - Permanent ventricular assist device support versus cardiac transplantation. AB - BACKGROUND: Although cardiac transplantation provides excellent therapy for some patients with terminal heart failure, the results are limited by the scarcity of donor organs, reduced long-term survival, and comorbid conditions. Current experience with temporary left ventricular assist devices suggest that a permanent, totally, or near totally implantable device may be a viable alternative. METHODS: We analyzed data from the 1997 International Society for Heart and Lung Transplantation (ISHLT) Registry and other literature on heart transplantation and compared survival and complication rates with our experience and that of others with temporary ventricular assist devices. From these data, we attempted to identify those patients who would benefit most from permanent left ventricular assist systems (LVASs). RESULTS: Among heart transplant candidates, United Network for Organ Sharing (UNOS) status II, O blood type, weight >180 lb, older age, and preformed antibodies are negative factors for receipt of donor hearts. Of patients transplanted, women and nonwhites have poorer outcomes. Success with wearable LVAS's suggest some of these patients might be better served with an LVAS than with cardiac transplantation. CONCLUSIONS: Because LVAS's could be made readily available without the need for a waiting list, they might compete well with the strategy of cardiac transplantation. PMID- 10475480 TI - Left ventricular assist system as a bridge to myocardial recovery. AB - BACKGROUND: Despite recent advances in medical therapy, heart transplantation, and mechanical circulatory support, the mortality of patients with congestive heart failure remains high. METHODS: Retrospective data on 5 patients were obtained from our hospital's medical records. Each patient was supported by a left ventricular assist system (LVAS) because of severe congestive heart failure. The duration of LVAS support averaged 229 days (range, 46 to 447 days). In 3 patients, the LVAS was removed electively after the patient showed recovery of myocardial function. In the other 2, it was removed because of a malfunction. RESULTS: In response to LVAS support, hemodynamic variables were significantly improved. The mean cardiac index increased from 1.45 to 2.69 L x min(-1) x m(-2) (p < 0.001) and the mean left ventricular ejection fraction increased from 0.144 to 0.288 (p < 0.025). All patients were in New York Heart Association functional class IV at LVAS implantation and class I at its explantation. One patient died of noncardiac-related causes 10 days after LVAS removal. The remaining 4 patients are alive and well 35, 33, 14, and 2 months after LVAS removal. CONCLUSIONS: In select patients with severe congestive heart failure, mechanical unloading with an LVAS can result in recovery of myocardial function. These patients can return to a normal physical status, thereby avoiding heart transplantation. More research is required to determine optimal modes of LVAS support, to predict which patients are likely to recover, and to assess long-term outcomes. PMID- 10475481 TI - Cardiac recovery in dilated cardiomyopathy by unloading with a left ventricular assist device. AB - BACKGROUND: Lasting recovery from intractable end-stage dilated cardiomyopathy, which occurs with ventricular unloading, has recently been demonstrated in 5 patients. Here our extended clinical experience with the "weaning" concept is presented. METHODS: In 19 patients (23 to 65 years) with intractable end-stage dilated cardiomyopathy, ventricular assist devices were explanted after support periods of up to 26 months, when repeat off-pump studies had shown either restoration of cardiac function (left ventricular ejection fraction, > 45%) and dimensions (left ventricular internal diameter in diastole, < 55 mm) or partial recovery (left ventricular ejection fraction between 35% and 40%) with serious complications on the device. At the time of device placement left ventricular ejection fraction was below 20% and left ventricular internal diameter in diastole more than 64 mm and bridge-to-transplantation had been planned. RESULTS: Seven patients with persistently restored cardiac function for more than 8 months and 5 patients for less than 5 months after weaning were studied. Five patients with recurrent heart failure died within 4 to 8 months after explantation. Four patients had to be transplanted and 2 died for reasons unrelated to cardiac function. An individual optimal left ventricular ejection fraction and left ventricular internal diameter in diastole was reached before pump removal was actually conducted in all patients. These parameters gradually deteriorated until pump removal. CONCLUSIONS: Lasting recovery can be reached by ventricular unloading in a subset of patients with intractable end-stage dilated cardiomyopathy. Obviously, there is an individual optimum of recovery that cannot be further improved by prolonged unloading. PMID- 10475482 TI - Dynamic cardiomyoplasty: at the crossroads. AB - BACKGROUND: Dynamic cardiomyoplasty remains a promising, but still unproven surgical treatment for patients with end-stage heart failure. Lack of a clear survival advantage and ongoing misunderstanding of its mechanism of action have hindered its acceptance as a treatment alternative for patients with end-stage heart failure. This review seeks to update current clinical results and practice of dynamic cardiomyoplasty and to present its likely mechanism of action. METHODS: The method involved a literature review. RESULTS: More than 600 patients have undergone dynamic cardioplasty since 1985. Improvement in average New York Heart Association class was noted in 80% to 85% of hospital survivors. Operative mortality has decreased from 31% in Phase I to less than 3% in the ongoing Phase III trial. Clinical work as well as recent animal work supports the hypothesis that through a combination of long-term elastic constraint and active dynamic assist, dynamic cardiomyoplasty decreases myocardial wall stress associated with the remodeling process of progressive heart failure. CONCLUSIONS: Though dynamic cardiomyoplasty can be shown to limit the remodeling process of heart failure in animal studies and some patients, its ultimate role in the treatment of heart failure will depend on the outcome of randomized, controlled studies. PMID- 10475483 TI - Arizona experience with CardioWest Total Artificial Heart bridge to transplantation. AB - BACKGROUND: We hypothesized that bridge to transplantation with the CardioWest Total Artificial Heart would succeed in a large percentage of patients. Further, we hypothesized that this success rate would not be significantly decreased by infection or thromboembolism. METHODS: From 1993 to March 1999, 24 patients received implants with the intention of bridge to transplantation. Data were collected prospectively. Heparin, coumadin, aspirin, ticlopidine, dipyridamole, and pentoxifylline were used for anticoagulation. RESULTS: Four patients died while on device support. Nineteen of 23 patients (83%) were transplanted. All 19 survived long term. One patient remains on CardioWest Total Artificial Heart support 6 weeks after implant. There was one stroke on the day of transplantation. There was a second stroke on the day of implantation. Neither stroke caused significant residual deficits. Both were in close relationship to an operative procedure. There were no serious device-related infections. CONCLUSIONS: The CardioWest Total Artificial Heart salvaged 20 of 24 critically ill patients. Neither infections nor neurologic problems were significant. We believe it is the device of choice for decompensating patients with biventricular failure who have adequate body and heart size. PMID- 10475484 TI - The Hemopump in 1997: a clinical, political, and marketing evolution. AB - BACKGROUND: The Hemopump (Medtronic, Inc, Minneapolis, MN) was conceived in 1975 and designed in 1982 as a temporary, extracorporeal cardiac assist system. Although it has been used clinically in Europe, it is not currently available in the United States. METHODS: In vitro and in vivo testing of the Hemopump began in 1983. Clinical investigations have included studies of patients in cardiogenic shock, Hemopump-supported coronary artery bypass operations in Sweden, and European studies of percutaneous transluminal coronary angioplasty (PTCA) with Hemopump support. RESULTS: The Hemopump has demonstrated positive hemodynamic effects in patients. Laboratory and clinical studies have shown that the nonpulsatile axial flow generates flows of up to 4.5 L/min while maintaining adequate perfusion of other organs. In Europe, hemopumps have been used successfully to support coronary bypass and PTCA. CONCLUSIONS: The Hemopump system is simple, inexpensive, and well tolerated by the blood elements. Moreover, its design allows flexibility in supporting patients during cardiopulmonary bypass (in lieu of conventional techniques) and high risk angioplasty, as well as in rescuing patients with low cardiac output. PMID- 10475485 TI - Heart booster: a pericardial support device. AB - BACKGROUND: Congestive heart failure is a pervasive disease afflicting millions of people. For many, their quality of life can be significantly improved by a pericardial device that can enhance cardiac output without the added risk of thromboembolism associated with direct blood contact. METHODS: A cardiac assist device with tubular elements is wrapped around the heart. Fluid is pumped into and out of the wrap causing contraction and dilation of its circumference. This contracting and relaxing action generates cardiac assistance without the need to contact blood. In vitro characterization and in vivo studies in calves were conducted to demonstrate the characteristics of the device. RESULTS: In vitro characterization with the device wrapped around one-half of a ventricle to simulate left ventricular support demonstrated outputs of 6.5 L/min at physiological afterloads. In vivo studies in calves demonstrated both cardiac output and afterload enhancements when the device is activated. CONCLUSIONS: This study provides a first demonstration of a device that provides cardiac support by a contraction and relaxation scheme with the device wrapped around the epicardium of the heart. The main feature of this actuation method is the potential for building a small device for implantation without blood contact. PMID- 10475486 TI - The AB-180 circulatory support system: summary of development and plans for phase I clinical trial. AB - BACKGROUND: The AB-180 circulatory support system is a small, durable, efficient centrifugal pump with low thrombogenic potential. The device was designed to provide a fully implantable, left ventricular assist system for short-term support to address the issues of systemic anticoagulation, thrombus formation, infection, and cost. METHODS: Extensive bench and animal studies were performed to validate the mechanical integrity of the device and its functionality as an implant. RESULTS: These studies demonstrated anticoagulation requirements, established operating guidelines, incorporated safety systems, and demonstrated safety and efficacy. CONCLUSIONS: The AB-180 fulfills the stated goals on initial evaluation. A phase I human trial is underway. PMID- 10475487 TI - Development of a totally implantable biventricular bypass centrifugal blood pump system. AB - BACKGROUND: During the past 2 years, the development of a totally implantable biventricular bypass rotary blood pump system has been made. METHODS: An extracorporeal gyro centrifugal pump, the CIE3, was miniaturized and developed into the PI601, a totally implantable plastic pump. Two-day anatomic and hemodynamic feasibility studies demonstrated that these two pump systems were easily implantable inside a calf's abdominal wall, directly under the diaphragm. The priming volume of the pump was 20 mL, with sufficient cardiac outputs at approximately 2,000 rpm and requiring less than 10 W of power. Two-week antithrombogenic screening tests also revealed these pump systems to be quite antithrombogenic. In addition, 1-month system reliability studies demonstrated fail-safe reliable performances. RESULTS AND CONCLUSIONS: Encouraged by these preliminary studies, the PI601 model was converted to the permanently implantable titanium gyro pump PI702 model. The long-term implantations were initiated approximately 3 months ago, and two such long-term LVAD studies are currently underway with no sign of difficulty (October 10, 1997). They were followed 283 days and 72 days, respectively. Both terminated due to functional inflow obstruction. There were no blood clots or emboli at autopsy. PMID- 10475488 TI - A ventricular assist device powered by conditioned skeletal muscle. AB - BACKGROUND: We are developing and testing a new ventricular assist device (VAD) to be powered by conditioned skeletal muscle. METHODS: To evaluate the VAD hardware and to develop a muscle training regimen, 8 calves have been used in studies in which the right latissimus dorsi muscle was employed. The experiments were carried out to an approximately 4-month duration. RESULTS: There was significant conversion of type II (fast twitch) to type I (slow twitch) muscle fibers. This did not correlate well, however, with device performance. The device stroke volumes ranged from approximately 17 to 90 cc. This variability of outcome occurred despite the fact that identical hardware, surgical procedures, and training regimens were employed. CONCLUSIONS: The results from the first eight studies lead us to speculate that perfusion may be important even when the muscle is working at pressures much lower than systemic blood pressure levels. In an attempt to augment tissue perfusion, we plan to investigate thermally induced angiogenesis as a possible mechanism for increasing blood flow to the tissue. PMID- 10475489 TI - Progress with the HeartSaver Ventricular Assist Device. AB - BACKGROUND: Ventricular assist devices (VADs) have been shown to be effective for short- or long-term circulatory support. Devices are either being adapted or newly designed for longer term or permanent support, with the goal to provide patients with improved quality of life. Since 1990, a program has been in place to develop a totally implantable, permanent VAD. METHODS: A multidisciplinary team is developing this VAD with specific goals in mind: (1) that it have an intrathoracic position, (2) that it be a totally implantable device without any percutaneous connections, and (3) that it be possible to communicate with the device from remote locations. These goals would allow for complete patient mobility and flexibility for follow-up. RESULTS: The electrohydraulically actuated VAD combines the blood pump, volume displacement chamber, energy converter, and internal electronic module into a single compact unit. The device called the HeartSaver VAD is powered by a transcutaneous energy transfer system and can be remotely monitored and controlled. Prototypes of different versions of the device have been tested in vitro and in vivo with satisfactory performance. CONCLUSIONS: The prototypes of the HeartSaver VAD have functioned well under test conditions and fulfilled the outlined goals. Further development and testing of the design are being conducted before clinical availability. PMID- 10475490 TI - Development of the Nimbus/University of Pittsburgh innovative ventricular assist system. AB - BACKGROUND: Nimbus Inc, and the University of Pittsburgh's McGowan Center for Artificial Organ Development have been collaborators on rotary blood pump technology initiatives since 1992. Currently, a major focus is an innovative ventricular assist system (IVAS) that features an implantable, electrically powered axial flow blood pump. In addition to the blood pump, a major development item is the electronic controller and the control algorithm for modulating pump speed in response to varying physical demand. METHODS: Methods used in developing the IVAS include computational fluid dynamic modeling of the pump's interior flow field, flow visualization of the flow field using laser-based imaging, computer simulation of blood pump-physiological interactions, vibroaccoustic monitoring, and an extensive in vivo test program. RESULTS: Results to date, which are presented below, include successful in vivo tests of blood pumps with blood immersed bearings, and feasibility demonstration of vibroacoustic monitoring in this application. CONCLUSIONS: This unique blend of industrial experience and technologies with the University-based Research and Development Center has greatly enhanced the progress made on this IVAS project. PMID- 10475491 TI - Midnight basketball for girls. PMID- 10475492 TI - Received social support for sexually transmitted disease-related care-seeking among adolescents. AB - OBJECTIVE: To describe the types and sources of social support received by adolescents obtaining care at a large urban sexually transmitted diseases (STD) clinic. METHOD: A total of 140 females and 82 males (ages 13-20 mean, 17.6 years) indicated whether they had received any of 11 types of social support, and, if so, from whom. Types of support included: companionship to clinic, advice on symptom interpretation, advice to seek clinical care, advice on potential sources of care, help making appointments, prior provision of medications, supportive talking, help talking to sex partner, provision of money, provision of transportation, and help getting STD protection. RESULTS: Eighty percent received at least one type of social support of whom 77% of subjects reported at least two types of support. The most frequently received support was information about symptom interpretation and appropriate clinic use (47% for each); 41% were accompanied to their clinic visit, and 37% received emotional support. A total of 15% of men but only 4% of women (p < .05 by Chi-square) received medication (usually antibiotics) but women were more likely to receive financial help (5% vs. 0% for women and men, respectively; p < .05). Although women obtained support earlier in the care-seeking process than men, there were no other significant gender differences in types of received support. Friends and sex partners were the most frequently cited sources of companionship and transportation, but parents provided transportation, information, medicine, and money for 15-20% of subjects receiving these types of support. CONCLUSION: Most adolescents receive a social support as part of seeking care for STD-related problems. PMID- 10475493 TI - Risky sexual behaviors among adolescents with alcohol use disorders. AB - Little is known about risky sexual activity among adolescents with alcohol use disorders. This study of 371 adolescent drinkers found that those with alcohol disorders were more likely than other drinkers to be sexually active, to have greater numbers of partners, and to initiate sexual activity at slightly younger ages. Independent of alcohol group, females were more likely than males to have unprotected sexual encounters. PMID- 10475494 TI - Changes in incarcerated adolescents' human immunodeficiency virus knowledge and selected behaviors from 1988 to 1996. AB - Data were collected from juveniles in Alabama and Florida correctional facilities. The samples were closely demographically matched. Analysis indicated significant increases in knowledge about human immunodeficiency virus between 1988 and 1996 and increased sexual risk behaviors. Knowledge levels were lower for ethnic minority groups and boys, suggesting ethnic- and gender-specific pedagogical aids are needed. PMID- 10475495 TI - HIV prevention in street youth. AB - Homeless adolescents have remained an underserved population throughout the human immunodeficiency/acquired immune deficiency syndrome epidemic. This article reviews the recent literature investigating human immunodeficiency virus (HIV) risk behavior among street youth. Prevalence rates of both adolescent homelessness and HIV seropositivity are unknown. However, data from a number of samples document a high prevalence of HIV risk behavior, sexually transmitted diseases, and alcohol/drug use among homeless adolescents. A number of individual and social factors, often associated with street survival, propel adolescents toward high-risk behavior. For some adolescents, testing HIV positive is perceived as advantageous in the procurement of basic needs such as food and shelter. HIV risk-reduction interventions must take into consideration the cause of homelessness, access to and participation in shelter services, and individual factors (such as the effects of sexual orientation and ethnicity) that frequently have not been systematically included in previous research. HIV risk for many homeless adolescents stems directly from their state of homelessness. National policies and funding are needed to address the health needs of these youth. PMID- 10475496 TI - Adolescents' recall of sexual behavior: consistency of self-report and effect of variations in recall duration. AB - PURPOSE: To describe the relationship between adolescents' 2-week, 2-month, and 12-month recall of sexual behavior; to assess the variability of adolescents' self-reported sexual behaviors over a period of 1 year; and to draw conclusions regarding the use of recall periods in measuring self-reported sexual behavior in adolescents. METHODS: Data from 296 African-American adolescents (age 12-19 years; 28% male) were analyzed. Baseline data comprise 2-week, 2-month, and 12 month recall of number of partners and frequency of condom-protected and unprotected vaginal, oral, and anal sex. Self-reported frequency of refusal of unprotected sex during the 2-week and 2-month recall periods are also included. To assess variability in self-reports of number of partners and frequency of behaviors over time, repeated measures of 2-week and 2-month recall were collected from a subset of the sample (n = 129; 24% male). RESULTS: The strength of correlation among responses from the three recall periods was dependent upon (a) the difference in length of the recall periods, and (b) the nature of the construct being recalled (e.g., number of partners vs. number of behaviors). Longitudinally, the variability of 2-week recall responses was generally larger than the variability in 2-month recall responses. CONCLUSIONS: Consistent estimates of adolescents' sexual behavior over a 1-year period may be obtained from several assessments of 2-week recall, or from relatively fewer assessments of 2-month recall data. PMID- 10475497 TI - High school athletic participation, sexual behavior and adolescent pregnancy: a regional study. AB - PURPOSE: To determine whether high school athletic participation among adolescents in Western New York was associated with reduced rates of sexual behavior and pregnancy involvement. METHODS: A secondary analysis of data from the Family and Adolescent Study, a longitudinal study of a random sample of adolescents (ages 13-16 years) from 699 families living in households in Western New York. A general population sample was obtained with characteristics closely matching the census distributions in the area. Interview and survey methods provided data on athletic participation, frequency of sexual relations during the past year, and risk for pregnancy. Bivariate correlations were used to examine relationships among athletic participation, demographic and control variables, and measures of sexual behavior and pregnancy rates. Next, path analyses were done in order to test for hypothesized relationships between athletic participation, sexual behavior, and pregnancy involvement while controlling for age, race, income, family cohesion, and non-athletic forms of extracurricular activity. Variables that were significantly associated with sexual behavior and/or pregnancy involvement were presented for both sexes within the resulting multivariate models. RESULTS: Lower income and higher rates of sexual activity were associated with higher rates of pregnancy involvement for both sexes. Family cohesion was associated with lower sexual activity rates for both sexes. For girls, athletic participation was directly related to reduced frequency of sexual behavior and, indirectly, to pregnancy risk. Male athletes did not exhibit lower rates of sexual behavior and involvement with pregnancy than male non-athletes. Boys who participated in the arts, however, did report lower rates of sexual behavior and, indirectly, less involvement with pregnancy. CONCLUSIONS: Female adolescents who participated in sports were less likely than their non-athletic peers to engage in sexual activity and/or report a pregnancy. Among male adolescents, athletic participation was unrelated to sexual behavior and pregnancy involvement. Teen pregnancy prevention efforts for girls should consider utilizing sport as a strategic tool. PMID- 10475498 TI - Subsequent infection among adolescent African-American males attending a sexually transmitted disease clinic. AB - PURPOSE: To identify predictors of subsequent infection among a sample of 15- to 19-year-old African-American males attending an urban sexually transmitted disease (STD) clinic in the Midwest. METHODS: During a 14-month period, 562 youth participated in a brief STD prevention intervention designed to promote condom use. They completed self-administered questionnaires (baseline, posttest, 30-day and 6-month) on their sexual and condom use behavior in the past month. Infection data (baseline, 5 years before, and 12 months after baseline) were obtained from clinic and state surveillance records. Logistic regression was used to predict infection within 6 and 12 months of the baseline visit. RESULTS: Within 12 months of the baseline, 31.3% were treated for an infection, of whom 1.4% returned within 30 days, an additional 17.1% within 6 months, and the remaining 12.8% within the last 6 months. The 12-month rate was 1.6-1.7 times higher than the rates reported for older STD clinic attendees. Subsequent infection was positively associated with age at first intercourse, number of children fathered, infection prior to and at the index visit, exchange of sex for drugs in the past year, and perceived risk of infection within the year; it was negatively associated with frequency of condom use with one's steady partner. CONCLUSIONS: Sexually transmitted disease clinic staff routinely obtain information from young African-American males that can be used to identify individuals who are most likely to become reinfected. Because repeaters account for a disproportionate number of infections, prevention efforts tailored to their needs would have a corresponding impact on STD rates. PMID- 10475499 TI - Cultural factors affecting diet and pregnancy outcome of Mexican American adolescents. AB - PURPOSE: To describe the nutrition knowledge, cultural beliefs, food intake, and attitudes about weight gain of Mexican American adolescents and their relationship to pregnancy weight gain and infant's birth weight. METHODS: A convenience sample of 46 pregnant adolescents, who were self-identified as Mexican American, primigravidas, and aged 13-18 years, was eligible. Two personal interviews were conducted with each participant, at the second trimester (18-22 weeks' gestation) and third trimester (30-34 weeks' gestation). The measurements were nutrient intake, nutrition knowledge, and attitudes toward weight gain. The proxy measure for acculturation was the length of residence in the United States [Group 1 (G1), n = 14; 3-12 months), G2 (n = 19; 12-48 months), and G3 (n = 13; 48-216 months). Descriptive statistics and analysis of variance were performed on the data as appropriate. Qualitative methods were used to describe cultural beliefs, behaviors, and attitudes during pregnancy. RESULTS: The mean age of the sample was 16.34 +/- 1.56 years. A total of 21.74% (n = 10) was American-born and 78.26% (n = 36) was Mexican-born. G3 was significantly (p < .05) the youngest at the time of conception, gained the most weight during pregnancy, was more knowledgeable about nutrition, was more educated, and was single and living with parents. There were no differences regarding the adequacy of diets during pregnancy among the three groups, and all diets adhered to up to 85% of the mean adequacy ratio (MAR). The mean total weight gain was 31.83 +/- 9.88 lb. There were no statistically significant differences of birth weight; the mean value for the three groups was 7.23 +/- 0.98 lb. CONCLUSIONS: The most powerful factors that reportedly contributed to good food practices during pregnancy were maternal concern about the well-being of the baby, role of motherhood, and family support system. It was found that with acculturation, adolescents lost most of the traditional Mexican cultural beliefs related to pregnancy, and their attitudes about weight gain were more negative. There was no relationship between nutrition knowledge and diet adequacy during pregnancy; they chose the traditional foods thought most nutritious by parents and family, for the benefit of their baby. PMID- 10475500 TI - Pseudocyesis in an adolescent using the long-acting contraceptive Depo-Provera. PMID- 10475501 TI - Outcome of external cephalic version after 36 weeks' gestation without tocolysis. AB - OBJECTIVE: Despite its efficacy, external cephalic version (ECV) at term is not universally employed. Published series of Caucasian women are small and concerns regarding safety and spontaneous version remain. We review the outcomes of 26 months of a breech clinic in a Dublin teaching hospital. METHODS: All women with known breech presentation at 36+ weeks were referred unless another indication for cesarean section existed. Unstable lie, fetal compromise, antepartum hemorrhage, and patient refusal were the only contraindications to ECV. One operator attempted all versions, without tocolysis. RESULTS: Three hundred seventy women were referred and in 356 (95%), version was attempted at a mean gestation of 37 + 3 weeks; 195 (55%) were nulliparous and 161 (45%) were multiparous. The success rate was 43%, including 33% of nulliparous and 54% of multiparous women. There were three perinatal deaths (0.8%), all unrelated to the version. Minor complications were rare, although two women were delivered by cesarean section shortly after unsuccessful version. Ninety-three percent of successful versions were cephalic at delivery, as were 4% of unsuccessful versions; 12% of infants with a cephalic presentation after successful version were delivered by cesarean section. CONCLUSIONS: From this large series, ECV is extremely safe. Spontaneous version after unsuccessful ECV and reversion after successful ECV are unusual. PMID- 10475502 TI - Fetal acoustic stimulation in early labor and pathological fetal acidemia: a preliminary report. AB - OBJECTIVE: To determine if a nonreactive response to fetal acoustic stimulation in early labor can predict a significantly higher risk of umbilical arterial pH <7.10 or <7.00. METHODS: Fetal acoustic stimulation was applied to the fetuses of term parturients (gestational age > or =37 weeks) with cervical dilation of < or =5 cm. The responses to stimulation were correlated with cesarean delivery for fetal distress and umbilical arterial pH. Student's t-test, Chi-square, and Fisher exact test were used; P < 0.05 was considered significant. Relative risks (RR) and 95% confidence intervals (CI) were calculated. RESULTS: The study population contained 271 subjects, of which 90% (244) had a reactive response following acoustic stimulation and 10% (27) a nonreactive response. The maternal demographics, time interval from stimulation to delivery (8.3 +/- 8.7 vs. 8.3 +/- 8.4 h; P = 1.00) were similar in the two groups. Compared to those with a reactive response, patients with a nonreactive response had a significantly greater risk for: 1) cesarean delivery for fetal distress (2.0% vs. 11.1%; P = 0.03, RR 4.1, 95% Cl 1.5, 60.5), 2) umbilical arterial pH <7.10 (2.0% vs. 14.8%; P = 0.007, RR 5.0, 95% CI 2.2, 11.6), and 3) umbilical arterial pH <7.00 (0.8% vs. 7%; P = 0.05, RR 5.0, 95% CI 1.8, 15.2). CONCLUSION: A nonreactive response to fetal acoustic stimulation in early labor is associated with a significantly increased risk for cesarean delivery for fetal distress and neonatal acidosis. This finding extends the potential value of acoustic stimulation as an intrapartum admission screening test. PMID- 10475503 TI - Matrix metalloproteinases-9 in preterm and term human parturition. AB - OBJECTIVE: Spontaneous rupture of the fetal membranes occurs after the commencement of labor in 90% of cases. Recent evidence indicates that the process of parturition requires not only an increase in myometrial contractility and cervical ripening, but also degradation of extracellular matrix in fetal membranes (i.e., leakage of fibronectin into cervico-vaginal secretions). This study was undertaken to determine if parturition is associated with in vivo evidence of increased bioavailability of matrix metalloproteinases-9 (MMP-9) and its inhibitor, tissue inhibitor of metalloproteinases-1(TIMP-1). METHODS: A cross sectional study was conducted with women in the following categories: 1) midtrimester (n = 25); 2) preterm labor and intact membranes in the absence of intraamniotic infection (n = 78); 3) term not in labor (n = 25); and 4) term with intact membranes in labor (n = 25). MMP-9, and TIMP-1 were measured using sensitive and specific immunoassays. RESULTS: 1) Spontaneous labor at term was associated with a significant increase in MMP-9 but not in TIMP-1.2) Women with preterm labor who delivered prematurely had significantly higher concentrations of MMP-9 but not TIMP-1 in amniotic fluid than those with preterm labor who delivered at term. 3) The concentrations of TIMP-1 decreased with advancing gestational age. In contrast, MMP-9 concentrations did not change with advancing gestational age. CONCLUSIONS: Spontaneous human parturition is associated with specific changes in the enzymatic machinery responsible for extracellular matrix degradation. PMID- 10475504 TI - Effects of preconceptual caffeine exposure on pregnancy and progeny viability. AB - OBJECTIVE: A previous study demonstrated for the first time that a drug such as caffeine, administered prior to ovulation and genomic activation, causes a quantitative difference in growth-promoting energy utilization in a proportion of 5-day-old blastocysts. The objective of the present study was to investigate whether developmental changes induced by caffeine administered throughout the estrus cycle prior to fertilization are sustained throughout pregnancy and after birth. METHODS: Caffeine was administered to rats throughout the estrus cycle prior to fertilization, with control and experimental groups subdivided into preimplantation and postimplantation categories. Preimplantation fertilization rate was assessed on day 4 of pregnancy by a pregnancy-induced elevation in maternal plasma progesterone concentration, or by flushing each uterine horn on day 5 of pregnancy to determine the presence or absence of a litter. Postimplantation fetuses were collected on gestational day 12 or allowed to go to term. RESULTS: Preconceptual caffeine exposure significantly reduced maternal fertility by the failure of a proportion of the litters to implant, rather than curtailing preimplantation development or postimplantation losses. Postnatal mortality between weeks 0 and 1 was elevated and the weekly incremental growth rate of the pups from week 3 through week 7 was significantly reduced in the preconceptually caffeine-treated offspring. Experimental females reached puberty at the same age as the controls but at a significantly lower body weight. Gestation length, hirthweight, litter size, sex ratio, and anogenital distance (a measure of prenatal androgenization) were not affected by preconceptual caffeine treatment. CONCLUSIONS: It was concluded that the reduced fertility rate in preconceptually caffeine-exposed rats was due to the failure of litters to implant rather than to a reduced fertilization rate, which was normal. It was further concluded that the growth rate over the neonatal and prepubertal periods of surviving pups in the caffeine-treated group was subnormal. PMID- 10475505 TI - Outcome of patients undergoing transabdominal cerclage: a descriptive study. AB - OBJECTIVE: Transabdominal cerclage is now evolving as an alternative to transvaginal cerclage in patients with cervical incompetence. The purpose of our study was to evaluate and describe our experience in patients selected to undergo transabdominal cerclage placement. METHODS: Outcome data for all patients who underwent transabdominal cerclage from January, 1990, through December, 1994, was collected. Indications for transabdominal cerclage included patients with prior failed vaginal cerclage, extremely shortened cervix, or anatomical defects judged unsuitable for transvaginal cerclage. RESULTS: Eleven patients underwent 12 transabdominal cerclage procedures. The mean gestational age at the time of cerclage placement was 12.5 weeks (range 10-14 weeks). The mean gestational age at delivery was 34 weeks. There were two neonatal deaths, which were related to extreme prematurity. The mean birthweight was 2,622 g. The fetal salvage rate was 83%. CONCLUSIONS: These findings suggest that in a highly select group of patients, the transabdominal cerclage can be a safe and effective procedure. PMID- 10475506 TI - High prevalence of activated protein C resistance due to factor V leiden mutation in cases of intrauterine fetal death. AB - OBJECTIVE: To test a possible association between activated protein C resistance and intrauterine fetal death. METHODS: The activated protein C anticoagulant activity and factor V R506Q mutation were assessed in 14 nonpregnant women with a history of intrauterine fetal death and 14 healthy controls. RESULTS: Four women in the study group were heterozygotes for the factor V mutation and none of the controls. The mean activated protein C activity of the study group was statistically significantly lower than that of the controls (P = 0.013). CONCLUSION: Resistance to activated protein C activity may be of etiologic importance in some cases of intrauterine fetal death. PMID- 10475507 TI - Congenital ischemic forearm necrosis associated with a compound presentation. AB - An infant born in a compound presentation had ischemic forearm necrosis requiring amputation. Bilateral renal vein thrombosis predating labor and delivery was also diagnosed. The possibility of congenital ischemic necrosis in an extremity should not affect the obstetrical management of compound presentation, but if this complication occurs a search for an underlying coagulopathy is important. PMID- 10475508 TI - Is the vacuum safe for preterm vaginal delivery? PMID- 10475509 TI - Allergic contact dermatitis in children and adolescents. AB - From a clinical point of view, the prevalence of allergic contact dermatitis (ACD) among children and adolescents seems to be low. However, many children have dermatitis, most often atopic dermatitis. In selected cases, ACD is suspected, and the child is tested. The question remains, whether the prevalence of ACD in children really is low or whether the possibility of ACD is not sufficiently considered. During the last decade, reports have appeared on series of children and adolescents with contact allergy and ACD. Few cases have been reported in infants, but the development of contact allergy and ACD increases with age. Most studies include selected groups of children and adolescents with suspected ACD. Few studies have examined unselected populations, and most consider only the prevalence of contact allergy without evaluating the clinical relevance, e.g., the prevalence of ACD. Furthermore, no follow-up studies exist. Therefore, the incidence and prevalence of contact allergy and ACD in children and adolescents is largely unknown. PMID- 10475510 TI - Laser Doppler perfusion imaging for the documentation of flare-up in contact allergy to gold. AB - A healed or healing allergic contact dermatitis is easily reactivated to a flare up if the patient is provoked systemically with his particular allergen. 5 patients with contact allergy to gold were patch tested with a serial dilution of gold sodium thiosulfate. 1 to 2 months later, when positive test reactions were healed, the patients were given 1 intramuscular injection of gold sodium thiomalate. This resulted in a flare-up of patch tests as well as a transient fever and toxicoderma-like reactions. The flaring tests were registered by digital photographs and laser Doppler imaging (LDI). An increased cutaneous blood flow in the formerly positive patch tests was discernible within 1 h after provocation, increased to a vigorous maximum after 4-6 h, and subsided during the following few days. The earliest increase and the highest peak of blood flow were observed in the test areas of the strongest original test concentration. LDI provides a most suitable technique for studying the dynamics of this rapidly developing inflammation of a mainly dermal character. PMID- 10475511 TI - Effects of various grit-containing cleansers on skin barrier function. AB - Products intended for individuals in contact with strongly adhering dirt often contain grit. Various clinical test methods have been developed for evaluating the potential of personal washing products to induce skin irritation. In the present study, differences in the irritant effects of washing products containing naturally-derived grit and synthetic grit were investigated in a forearm wash test. The forearms of 16 test subjects were washed in a total of 18 treatments (4 per day for 4 days, with 2 treatments on the 5th day). Treatment consisted of continuous washing for 2 min by a technician, who gently slid his fingertips with the lather up and down the forearm. Non-invasive instrumental measurements of skin barrier function were performed. Repetitive washing for 1 week lead to increased TEWL values, skin redness and decreased stratum corneum hydration. Results indicate differences in irritancy potential due to different types of grit, their surface and concentration. It is concluded that the repeated wash test seems to be adequate for rating personal washing products that contain grit. PMID- 10475512 TI - Hypersensitivity syndrome due to 2 anticonvulsant drugs. AB - Drug-induced hypersensitivity syndrome is a multiorgan-system reaction characterized by fever, pleomorphic eruption, lymphadenopathy, eosinophilia, lymphocytosis and hepatitis. We report a drug hypersensitivity syndrome in a 6 year-old Tunisian child treated for epileptic absences with sodium valproate and ethosuximide. Imputability of these 2 drugs is probable because of the chronological and clinical features. Positive rechallenge with ethosuximide confirmed the toxicity of this drug. Sodium valproate was also responsible because patch testing was positive and followed by a generalized eruption. Human herpesvirus 6 (HHV6) antibody titers increased significantly within 15 days. There was a favourable outcome after discontinuation of the drugs and corticosteroid therapy. Our case is interesting because this drug hypersensitivity syndrome occurred with non-aromatic anticonvulsant drugs. It is the 1st case with ethosuximide and the 2nd with sodium valproate. We also observed a reactivation of HHV6 infection that may contribute to the development of this hypersensitivity syndrome. PMID- 10475513 TI - Cross-reactivity patterns of palladium and nickel studied by repeated open applications (ROATs) to the skin of guinea pigs. AB - Cross-reactivity is usually studied with patch test techniques, but the relevance of a single 1-2 day exposure under occlusion can be questioned. To study relevance, animals were induced with PdCl2 or NiSO4 according to the guinea pig maximization test method and then treated for 10 days according to the repeated open application test (ROAT) method. Animals induced with PdCl2 reacted in the ROATs to PdCl2 (100%) but rarely to NiSO4. Animals induced with NiSO4 reacted in ROATs to the same degree with NiSO4 and PdCl2 (23-30%). The concordance between pre-ROAT patch test results and ROAT outcome was high for PdCl2 (100%) and low (10-40%) for NiSO4. Patch testing seems to overestimate the risk of skin reactions when guinea pigs sensitive to PdCl2 are treated topically with NiSO4. The finding from patch test studies that animals induced with NiSO4 react only to NiSO4 but not to PdCl2 was not confirmed. Repeated open applications more adequately mimic exposure conditions than does patch testing. PMID- 10475514 TI - Reproducibility of irritant patch test reactions to sodium lauryl sulfate in a double-blind placebo-controlled randomized study using clinical scoring. Results from a study group of the German Contact Dermatitis Research Group (Deutsche Kontaktallergie-Gruppe, DKG). AB - Reading of doubtful patch test reactions can be improved by comparing them to defined negative, allergic and irritant controls. For the latter, an irritant patch test is needed that gives sufficiently reproducible results. In our double blind, placebo-controlled, randomized study, we have analysed the synchronous reproducibility of patch tests with 0%, 0.0625%, 0.125%, 0.25%, 0.5% and 1.0% sodium lauryl sulfate (SLS). Tests and readings were done according to ESCD guidelines. 139 patients (75 women, 64 men) 18 to 77 years old were tested. The % of positive patients as well as the % of reproducible positive reactions increased with rising concentrations of SLS to a maximum of approximately 90% and 85%, respectively, with 1% SLS. Time courses of reactions were also related to the concentration of SLS: with 1% SLS, plateau-type time courses were most common (65%), followed by decrescendo- (23%) and crescendo-type reactions (12%). With 1% SLS, very similar mild to moderate reactions were obtained on days 2 and 3. The reaction threshold and reproducibility of tests were not related to age and sex of patients or to their own assessment of skin irritability. We conclude that 1% SLS is appropriate for an irritant patch test that gives sufficiently reproducible results. PMID- 10475515 TI - In vitro lymphocyte reactivity to gold compounds in the diagnosis of contact hypersensitivity. AB - The use of the lymphocyte transformation test (LTT) in the diagnosis of contact hypersensitivity to gold was studied in 8 patients who had positive patch tests to gold salts, and in 8 control subjects who were negative to such patch tests. Gold sodium thiosulfate and gold chloride were added to cultures of lymphocytes, which were labeled by 3H-thymidine after 96 h. The lymphocyte stimulation index was calculated as the beta-counts in stimulated cultures divided by those in control cultures. The index was statistically significantly higher for the patient group (p=0.005-0.04) than for the control group. Levels of interferon gamma (IFN-gamma) were determined for the supernatants of the lymphocyte cultures. An index IFN-gamma, which is defined as the level of IFN-gamma in stimulated cultures divided by that in control cultures, was statistically significantly higher for the patient group (p=0.01-0.006). The LTT stimulation index showed specificity and sensitivity between 67 and 80%, the respective values for Index IFN-gamma being between 73 and 100% when the patch test was used as a reference method. Evaluation of lymphocyte reactivity might be of future interest in the diagnosis of allergic reactions to gold if the sensitivity and specificity can be improved. PMID- 10475516 TI - Evaluation of the fragrance mix in the European standard series. PMID- 10475517 TI - Pentaerythritol-esterified gum rosin as a sensitizer in Granuflex hydrocolloid dressing. PMID- 10475518 TI - Occupational contact dermatitis caused by polyfunctional aziridine crosslinker: duct tubing for airconditioning. PMID- 10475519 TI - Occupational allergic contact dermatitis eczema caused by obeche and teak dusts. PMID- 10475520 TI - Occupational allergic contact dermatitis from N,N-methylene-bis-5-methyl oxazolidine in a nylon spin finish. PMID- 10475521 TI - Allergic contact dermatitis from dithranol. PMID- 10475522 TI - Active sensitization to sodium metabisulfite in hydrocortisone cream. PMID- 10475523 TI - Contact allergy to clotrimazole: an unusual allergen. PMID- 10475524 TI - Prevalence of natural rubber latex allergy in hairdressers. PMID- 10475525 TI - Allergic contact dermatitis from dithio-2,2-bis(benzmethylamide). PMID- 10475526 TI - Contact urticaria and systemic anaphylaxis from codfish. PMID- 10475527 TI - Occupational protein contact dermatitis from glucoamylase. PMID- 10475528 TI - Protein contact dermatitis from midge larvae (Chironomus thummi thummi). PMID- 10475529 TI - Exposure-pattern dermatitis due to 2-aminothiophenol and 2-aminophenyldisulfide. PMID- 10475530 TI - Exposure-pattern dermatitis caused by N-(2-amino-4,6-dichloro-5-pyrimidinyl) formamide. PMID- 10475532 TI - All the evidence points to alcohol being implicated in the recent changes in mortality in Russia. PMID- 10475531 TI - Alcohol and mortality in Russia: is it different than elsewhere? AB - PURPOSE: To discuss the hypothesis that alcohol, and binge drinking in particular, is a major determinant of the recent mortality fluctuations in Russia. METHODS: Discussion based on published literature. RESULTS: The hypothesis is based on circumstantial evidence. The changes in mortality coincided with introduction and collapse of the Soviet anti-alcohol campaign. The largest relative changes in mortality were observed for "alcohol-related causes" (a specific diagnostic category used in Russia) as well as violent and accidental deaths, but the largest absolute changes were observed for cardiovascular causes which had the largest impact on all-cause mortality. There is no direct support for the hypothesis. Available estimates of alcohol consumption in Russia are low, and the only published study on alcohol and mortality conducted in Russia produced negative results. Increase in drinking prevalence alone would not explain the mortality rise; increase in relative risk related to alcohol would also be needed. The biological mechanisms which could underlie the presumed strong effects of alcohol on heart disease are not clear. On the other hand, binge drinking has not been addressed adequately by research so far. CONCLUSIONS: Until a well designed study is conducted in Russia, the hypothesis remains debatable. PMID- 10475533 TI - Alcohol consumption and disability pension among middle-aged men. AB - PURPOSE: To analyze the relation between alcohol consumption and the risk of disability pension among middle-aged men. METHODS: In the mid-seventies, complete birth-year cohorts of middle-aged male residents in Malmo, Sweden, were invited to participate in a general health survey. The 3751 men with complete data who constituted the cohort in this study were followed for 11 years. Alcohol consumption was estimated from the scores obtained from a test designed to identify subjects with alcohol related problems. RESULTS: Of the 498 men granted disability pension during follow-up, 48 stated to be teetotalers. The cumulative incidence of disability pension among teetotalers was 19%, whereas, it was 12% and 16%, respectively, among men with low and high alcohol consumption. The adjusted relative risk (RR) for acquiring a disability pension (using the group with low alcohol consumption as reference) was 1.8 among abstainers and 1.3 among men with high alcohol consumption. CONCLUSIONS: Alcohol overconsumption, as well as teetotalism, showed a positive relation to disability pension, and a moderate alcohol intake was found to be beneficial with respect to the risk of future disability pension. PMID- 10475534 TI - Changing conditions and treatments in a dynamic cohort of ambulatory HIV patients: the HIV outpatient study (HOPS). AB - PURPOSE: Most HIV-infected persons are now treated as ambulatory patients. Obtaining continually updated data about these patients' changing conditions, therapies, and reimbursement is essential to health care provision and planning. The systematic tracking of patient medical and laboratory information in an ongoing commercial data collection program (The Health Research Network) allows clinicians to better understand health outcomes, practice patterns, and epidemiologic trends for their patients. METHODS: To evaluate trends in conditions and therapies of ambulatory HIV-infected patients, we analyzed such data electronically and prospectively collected in the HIV Outpatient Study (HOPS) from 1992 through 1996 from 1876 patients seen in 11,755 clinic visits to ten HIV clinical practices. RESULTS: Patients were as likely to be diagnosed with Mycobacterium avium complex ([MAC] 5.4 cases per 100 person-years) or wasting syndrome (7.8 cases per 100 person-years), as Pneumocystis carinii pneumonia ([PCP]; 7.6 cases per 100 person-years) or Kaposi sarcoma ([KS]; 6.9 cases per 100 person-years). A nested analysis showed that HIV-infected cigarette smokers were at substantially greater risk of pneumonia (relative hazard [RH] = 2.3), bronchitis (RH = 1.7) and hairy leukoplakia (RH = 1.9) than nonsmokers. By 1996, 35 (56%) of 62 patients with PCP, 9 (30%) of 30 patients with other pneumonias, 28 (90%) of 31 patients with KS, 35 (73%) of 48 patients with MAC, and 24 (63%) of 38 patients with cytomegalovirus retinitis were treated without hospitalization. CONCLUSIONS: The HOPS provides continually updated information on the changing characteristics, conditions, and therapy of ambulatory HIV infected patients. PMID- 10475535 TI - Serum ascorbic acid and cardiovascular disease prevalence in U.S. adults: the Third National Health and Nutrition Examination Survey (NHANES III). AB - PURPOSE: To examine the relation between serum ascorbic acid concentration, which reflects dietary and supplement intake, and the prevalence of cardiovascular disease. METHODS: We analyzed data from 7658 men and women enrolled in the Third National Health and Nutrition Examination Survey (NHANES III). We calculated odds ratios and 95% confidence intervals (CI) to estimate the relative prevalence of cardiovascular disease, defined as self-reported angina, myocardial infarction, or stroke. Because we detected an interaction between serum ascorbic acid concentration and alcohol intake, we performed analyses stratified by drinking status. RESULTS: Among participants who reported no alcohol consumption, serum ascorbic acid concentrations were not independently associated with cardiovascular disease prevalence. Among participants who consumed alcohol, serum ascorbic acid concentrations consistent with tissue saturation (1.0-3.0 mg/dl) were associated with a decreased prevalence of angina (multivariate odds ratio (OR): 0.48; 95% CI: 0.23% to 1.03; p for trend = 0.06), but were not significantly associated with myocardial infarction or stroke prevalence. CONCLUSIONS: These results suggest the possibility of a biologic interaction between ascorbic acid and alcohol and that higher intakes of ascorbic acid may be associated with a decreased risk of angina among drinkers. PMID- 10475536 TI - Work and leisure time physical activity and mortality in men and women from a general population sample. AB - PURPOSE: The purpose of this study was to examine the long-term relationships between total physical activity and mortality from all causes and coronary heart disease (CHD) in the general population. METHODS: A prospective design was used, following participants for 29 years, beginning in 1960. The study population consisted of a randomly selected sample of white male (n = 698) and female (n = 763) residents of Buffalo, New York with a 79.0% participation rate and follow-up rates of 96.0% and 90.2% in males and females, respectively. At baseline, comprehensive information was obtained regarding participants' usual physical activity at work and during leisure time. RESULTS: As of December 31, 1989, three hundred and two (43.3%) men and 276 (41.0%) women died, 109 (15.6%) and 81 (10.6%) from CHD, respectively. In men, a significant interaction was found between activity and body mass index (BMI) for both outcomes. In women, a significant activity by age interaction was observed. In non-obese men (BMI<27.02), activity was inversely associated with all-cause [relative risk (RR) = 0.59; 95% confidence interval (CI), 0.39-0.89] and CHD mortality (RR = 0.39; 95% CI, 0.18-0.83), independent from the effects of age and education. No such associations were found in obese men and increased risks could not be ruled out. Among women, younger participants (age <60 years) had a significantly reduced risk of CHD death with increased activity (RR = 0.26; 95% CI, 0.07-0.99). No other significant associations were observed. CONCLUSIONS: Physical activity favorably influences mortality risks in non-obese men and younger women. Gender specific factors should be considered for potential effect modification. PMID- 10475537 TI - Women, employment status, and hypertension: cross-sectional and prospective findings from the Atherosclerosis Risk in Communities (ARIC) Study. AB - PURPOSE: This study examined the cross-sectional and prospective associations between employment status and hypertension among middle-aged, African-American (AA) and European-American (EA) women participating in the Atherosclerosis Risk in Communities Study. METHODS: Employed women and homemakers from the baseline examination (1987-89) were included in the cross-sectional study (n = 7351). Associations between employment and the incidence of hypertension ascertained at visit 2 (1990-92) were determined among those who at baseline, had low-normal blood pressure (not hypertensive and systolic blood pressure (SBP) < or = 120 mm Hg systolic and diastolic blood pressure (DBP) < or =80 mm Hg (n = 3194). Logistic regression analysis was used to examine the association between employment status and hypertension by ethnicity, taking into account covariates. RESULTS: At baseline, employed women were less likely to be hypertensive (SBP > or =140 mm Hg or DBP > or =90 mm Hg or current use of antihypertensive drugs) than were homemakers (prevalence odds ratio) (POR) = 0.70; 95% confidence interval (CI) = 0.62-0.79), controlling for age, body mass index, and education. Among the subgroup who had low-normal blood pressure at baseline, employed women were less likely to develop hypertension during the three-year time period than were homemakers (odds ratio (OR) = 0.68; 95% CI = 0.44-1.05). The inverse association was stronger among AA (RR = 0.37; 95% CI = 0.16-0.88) than EA (OR = 0.83; 95% CI = 0.50-1.38) women. CONCLUSIONS: These findings suggest that the inverse association between hypertension and employment status is not due to a healthy worker effect, and that employment may confer protection against incident hypertension in women. PMID- 10475538 TI - Cancer prevention among urban southwestern American Indian women: comparison to selected Year 2000 national health objectives. AB - PURPOSE: The health issues which have historically faced American Indians are related to infectious disease, but today many chronic diseases play a large role in the health status of this special population. For example, existing data indicate that American Indians have the poorest cancer survival of any group in the U.S. (34% vs. 50% for U.S. Whites). Regular participation in cancer screening and lifestyle factors such as smoking, diet, and alcohol have been found to significantly contribute to cancer risk. However, available information about these behaviors is sparse and suffers from limitations. The purpose of this study was to assess the prevalence of cancer risk behaviors and provider recommendation for cancer screening among urban American Indian women. METHODS: A random household cross-sectional survey was administered to 519 adult American Indian women in Phoenix, Arizona. Smoking and weight status, alcohol consumption, physical activity, and provider referral were assessed by self-report. RESULTS: The results indicate that weight (69.6% were overweight) and excessive alcohol consumption (12.1 reported chronic drinking) were significant health issues in this population. The prevalence of provider referral for smoking cessation (14.9% among ever smokers) and breast as well as cervical cancer screening (30.1% and 55.5%, respectively) were below the 75% level set forth in the Year 2000 Goals. CONCLUSIONS: The results of this study suggest that increased cancer prevention and control efforts are needed in this special population to address the link between diet, weight, and long-term disease prevention. PMID- 10475539 TI - Neonatal tumors. Introduction. PMID- 10475540 TI - Neuroblastoma in the neonate. AB - Neuroblastoma is the most common malignant neoplasm occurring in the neonate and arguably the most variable in its presentation. This review examines differences in the tumor's clinical course in the context of known biologic determinants of behavior. Tumors that would go undetected except for prenatal ultrasonography or postnatal neuroblastoma screening characteristically undergo spontaneous regression or maturation and require little or no treatment. Most low stage tumors with favorable biological features are effectively treated with surgery alone. The more bulky locoregional (stage 3) tumors require chemotherapy in addition to surgery. The most challenging tumors are those that are disseminated at birth (stages 4 and 4S). Management plans for these tumors are based on the biological characteristics of the tumor and (in the case of stage 4S tumors) on the presence and extent of functional embarrassment of vital organs. PMID- 10475541 TI - Congenital and neonatal leukemia. AB - Congenital and neonatal leukemia occur rarely, yet carry high mortality rates and pose special problems for the perinatologist and hematologist. Although the etiology is unknown, the presence of leukemia at birth suggests genetic abnormalities and possibly intrauterine exposures to drugs or other toxins as contributing factors. Specific chromosomal rearrangements that are common in congenital leukemia have recently been identified and promise to enhance our understanding of these enigmatic diseases. The differential diagnosis is broad and includes many disorders that occur frequently in the neonatal period. Infants diagnosed with congenital or neonatal leukemia require thorough investigative workup and extensive supportive care. Although the prognosis is poor, recent use of high-intensity multiagent chemotherapy regimens has produced promising results. PMID- 10475542 TI - Neonatal brain tumors: a review. AB - This article presents two cases of infants with brain tumors and reviews the literature pertinent to congenital and neonatal brain tumors. Information regarding epidemiology, presentation, prognosis, and clinical management are also addressed with specific regard to differences between neonatal and childhood brain tumors. An appeal is made to consider (1) coordination of the care of these children through pediatric multidisciplinary neuro-oncology programs; (2) enrollment, whenever possible, in clinical trials; and (3) submission of available tumor tissue to pediatric tumor banks to assure its availability to interested researchers. PMID- 10475543 TI - Soft tissue sarcomas of infancy. AB - Malignancy occurring during the neonatal period (defined as the first 28 days of life) is over 3 times the incidence of other pediatric age groups. Of all neoplasia occurring in infants, benign and malignant, 25% are soft tissue tumors. Differentiating the benign lesions from the 15% that are malignant can be difficult. This article discusses the epidemiology, differential diagnosis, evaluation, and treatment of infants with soft tissue sarcomas. Fibrosarcoma and rhabdomyosarcoma are also discussed at length. The authors review other rare tumors as well. The impact on diagnosis of molecular techniques is included when appropriate. A multidisciplinary team approach for treatment of these infants is recommended. PMID- 10475544 TI - Wilms' tumor and related abnormalities in the fetus and newborn. AB - Mesoblastic nephroma, a benign tumor, is the most common renal neoplasm in neonates. Wilms' tumor (WT) may occur in newborn infants, but is more common in older children. The molecular genetics of WT involves one or more genes located on Chromosome #11 and probably other locations not yet elicidated. Germline mutations cause less than 5% of WTs; most WTs are sporadic. Precursor lesions to WT called nephrogenic rests may be detected before evolution to WT by imaging studies. Developmental anomalies comprising several different syndromes are associated with nephrogenic rests and predisposition to WT. Prospective surveillance for WT may be feasible in high risk infants identified on the basis of physical findings followed by testing for predisposing gene defects and periodic imaging of the kidneys and other organs at risk until the period of risk has ended. PMID- 10475545 TI - The histiocytoses of infancy. AB - The histiocytoses comprise a rare and incompletely understood group of diseases that are characterized by an abnormal proliferation of histiocytes, mononuclear phagocytic, and antigen-presenting cells belonging to the reticuloendothelial system. The two major types of histiocytoses, which often present during infancy and require significant medical attention, are Langerhans cell histiocytosis and primary hemophagocytic lymphohistiocytosis. Their clinical manifestations frequently overlap, complicating their distinction, but a prompt and accurate diagnosis is essential to deliver the optimal treatment and maximize the chances for a favorable outcome. For Langerhans cell histiocytosis, careful risk stratification is critical for the appropriate administration of therapy. Patients with good prognostic factors may need only observation as their disease spontaneously regresses or minimal intervention. Poor prognostic factors mandate more intensive treatment. Patients with primary hemophagocytic lymphohistiocytosis require chemotherapeutic induction of their disease into a state of remission followed by stem cell transplantation, which currently offers the only known cure. Improvements on current therapies for the histiocytoses will depend on continued advances in the understanding of these enigmatic diseases. PMID- 10475546 TI - The infant with a vascular tumor. AB - Vascular birthmarks look alike in various tints of blue, pink, or red. These anomalies are either vascular tumors or vascular malformations. Hemangioma is the most common vascular tumor of infancy, typically appearing in the neonatal period. Hemangiomas grow rapidly during the first year of life and regress slowly during childhood. Treatment is indicated for problematic or endangering hemangiomas, usually the administration of corticosteroids or interferon-alfa, and in some instances, embolization or surgical resection. The more aggressive and rare vascular tumors, kaposiform hemangioendothelioma and tufted angioma, are often associated with thrombocytopenia and coagulopathy (Kasabach-Merritt phenomenon) and less responsive to pharmacological therapy. Vascular tumors, particularly hemangioma, are often confused with vascular malformations; although etiology, histology, and clinical behavior are quite different. Vascular lesions should be biopsied if there is any suspicion of malignancy. PMID- 10475547 TI - Cystic hygroma in the fetus and newborn. AB - Cystic hygromas are developmental abnormalities of the lymphoid system that occur at sites of lymphatic-venous connection, most commonly in the posterior neck. They are frequently associated with karyotypic abnormalities, various malformation syndromes, and several teratogenic agents. The disease course of an infant with cystic hygroma is unpredictable. When diagnosed prenatally, the overall prognosis is poor. Cystic hygroma diagnosed after birth is usually associated with a good prognosis. This article reviews the embryologic, genetic, and pathologic correlates of these lymphatic system abnormalities, as well as the clinical course and outcome of the fetus and newborn with a cystic hygroma. Management strategies are reviewed, including newer nonsurgical therapies for the neonate with a cystic hygroma. PMID- 10475548 TI - Reoperation results after segmental prosthetic replacement of bone and joint for limb salvage. AB - Thirty-eight patients who had undergone reoperations because of complications or implant failure subsequent to a prosthetic replacement for limb salvage procedure for the treatment of musculoskeletal neoplasm were followed until death or a minimum of 2 years, then reviewed. The mean follow-up time before reoperation was 35 months, and the mean follow-up time after reoperation was 51 months. The major indication for reoperation was aseptic loosening (34%). The reoperative procedures included 19 major revisions of the prosthesis and 19 other reconstructions. There were 7 complications, including 1 local tumor recurrence. Twelve patients died of metastatic disease at a mean of 16 months after reoperation. The probability for patient survival was 74% beyond 3 years and 67% after 5 years. The surgical staging, site of tumor involvement, and type of reoperation all correlated with long-term patient survival. Overall, 72% of the patients achieved excellent or good functional results: a rate that was slightly inferior to that of the initial arthroplasty. Despite this, 84% of the implants demonstrated either excellent or good performance at 5 years. Therefore, reoperation because of a failed initial prosthesis or for other reasons after a limb salvage procedure using custom-designed or modular-segmental bone and joint implants seems feasible with no significant effect on subsequent functional performance or on patient survival. PMID- 10475549 TI - Economic evaluations in the hip arthroplasty literature: lessons to be learned. AB - Readers are increasingly encountering articles dealing with health economic evaluations that compare various surgical strategies, leaving orthopaedists with the challenge of determining which program is cost-efficient and truly pertains to their setting. This study carries out a systematic review of the literature to appraise the quality, quantity, and type of economic evaluation as it pertains to the hip arthroplasty literature. To identify all relevant articles, we conducted a comprehensive computerized bibliographic search of Medline from 1966 to 1996. This search produced 1,611 abstracts that were screened. Studies that were incorporated met the following inclusion criteria: i) formal economic analysis, ii) an intervention specific to hip arthroplasty, and iii) the perspective of the study was evident (ie, patient, provider, society). These studies were appraised with regards to methodologic soundness based on 8 established economic principles. Only 68 articles from the 138 retrieved met the study criteria. Only 2 of the 68 articles met all 8 criteria of a comprehensive economic evaluation. The hip arthroplasty literature is deficient in methodologically sound economic evaluations. Several guidelines are introduced to aid orthopaedists in appraising the various economic studies, and recommendations are made to improve the quality of these studies in the orthopaedic literature. We suggest that the generation of such information should rank high on the priority list of the orthopaedic profession, granting agencies, and governments. PMID- 10475550 TI - Total knee arthroplasty for steroid-induced osteonecrosis. AB - There have been only a few reports that evaluate the outcome of total knee arthroplasty in patients with steroid-induced osteonecrosis of the knee. We retrospectively reviewed 31 total knee arthroplasties in 24 patients with confirmed steroid-induced osteonecrosis of the knee. The average follow-up was 64 months. Of surviving knees, 92% had significant improvement in knee scores. Five knees (16%) required a revision procedure. Reasons for revision were aseptic loosening in 3 knees and sepsis in 2 knees. Complications not requiring revision occurred in 6 of 31 knees (19%). Survivorship of total knee arthroplasty to revision in patients with steroid-induced avascular necrosis of the knee was 84% at 5 years. Although there was a slightly higher complication rate, this may, in part, be due to the severity of the patients' underlying disease processes. Total knee arthroplasty can be a successful procedure for chronically ill patients with steroid-induced osteonecrosis. PMID- 10475551 TI - Primary total hip arthroplasty with a cementless porous-coated anatomic total hip prosthesis: 10- to 12-year results of prospective and consecutive series. AB - We performed a prospective study in 108 consecutive patients (116 hips) who were followed for a minimum of 10 years (10-12 years) after primary total hip arthroplasty using an uncemented porous-coated anatomic (PCA) hip prosthesis. The average age of the patients at operation was 48.4 years (range, 19-85 years), and the diagnosis was avascular necrosis of the femoral head in 46 hips, neglected femoral neck fracture in 27, osteoarthrosis secondary to childhood pyogenic arthritis in 24, childhood tuberculous arthritis in 5, and miscellaneous in 14. The average preoperative Harris Hip Score was 55 points, which improved to 87 points at 11 years. Seventy-five hips (65%) were excellent, 11 (9%) were good, and 30 (39%) were poor. The overall rate of revision was 15% (17 of 116 hips). The rate of revision of the femoral component was 11% (13 of 116 hips), and the rate of revision of the acetabular component was 15% (17 of 116 hips). The prevalence of thigh pain was 28% at 11 years. The increase in the incidence of aseptic loosening of the femoral component was found to explain the high incidence of severe thigh pain at 11 years' follow-up. At 11 years, there was femoral osteolysis in 69 hips (59%) and acetabular osteolysis in 65 hips (56%). At 6 years, 20 hips (17%) showed definite wear of the polyethylene liner. At 11 years, 81 hips (70%) showed definite wear of the polyethylene liner. Because the complication rate of the PCA hip prosthesis with respect to loosening, osteolysis, and excessive wear in the polyethylene liner is high, we abandoned the use of this implant. PMID- 10475552 TI - Efficacy of irrigation for removal of particulate debris after cemented total knee arthroplasty. AB - We studied the amount of particulate debris removed with pulsatile lavage irrigation before and after component implantation in 13 consecutive patients undergoing primary cemented total knee arthroplasty (TKA) done by a single surgeon. Before component implantation, the knees were copiously lavaged with 3 L of pressurized irrigant; all fluid was collected in 1 aliquot using standard wall suction canisters. After cementing the components in place, another 3 L of pressurized irrigant was used; this fluid was collected in 3 sequentially labeled 1-L aliquots. Collected fluids were centrifuged, and the residue was washed, recentrifuged, and dried. Residual particulate debris was quantitated by weight. An average of 537 mg/L (range, 16-1,406 mg/L) of debris were removed before implantation with 3 L of irrigation. An average of 217 mg/L (range, 31-999 mg/L), 52 mg/L (range, 0-189 mg/L), and 49 mg/L (range, 1-185 mg/L) of debris was removed after implantation with each of the additional liters. Using analysis of variance testing, there was a statistically significant difference between the amount of debris removed with 3 L and after 4 L (P = .02) and 5 L (P = .03) of irrigant. There was no statistical difference between irrigation with 5 L and 6 L of irrigation (P = .92). The residua particulate debris was also analyzed to determine the relative amounts of bone-soluable organics and polymethyl methacrylate (PMMA). Before implantation, the residual debris, by weight, consisted of 79% bone and 21% soluable organics. We found on average that after implantation of components the specimens contained 53% bone and 47% PMMA and soluable organics by weight. We believe that despite careful implantation and meticulous cement technique, large amounts of debris, including bone and PMMA, remain after TKA, which require at least several liters of pulsatile lavage to remove. Removal of this particulate debris may decrease third-body polyethylene wear. PMID- 10475553 TI - Humeral hemiarthroplasty of the elbow joint in young patients with rheumatoid arthritis: a report on 7 arthroplasties. AB - This article evaluates the early results of 7 humeral hemiarthroplasties of the elbow joint in 5 female rheumatoid patients using only the humeral component of the capitellocondylar prosthesis. The follow-up period was 25 to 109 months. All patients were 50 years old or younger; 3 patients were between 22 and 26 years. Because all patients were rather young, the intention was to perform a procedure that conserves a maximal amount of bone stock for future salvage procedures. One prosthesis had to be removed because of an infection unrelated to the primary procedure. The remaining bone stock provided a stable and pain-free excision arthroplasty. The other arthroplasties showed good pain relief; however, postoperative motion was not as reliable as described for total capitellocondylar prostheses. PMID- 10475554 TI - The Insall-Burstein total knee replacement in osteoarthritis: a 10-year minimum follow-up. AB - A total of 99 Insall-Burstein posterior stabilized (IBPS) knee replacements were implanted in 86 osteoarthritic patients. We reviewed 60 knees with a 10- to 15 year follow-up. Using the Knee Society score, 35 knees (58%) were excellent, 15 (25%) good, 4 (7%) fair, and 6 (10%) poor. Flexion at follow-up was 106 degrees on average. Moderate patellofemoral crepitation was present in 5 knees (9%), and 11 knee required excision of a synovial nodule proximal to the patella. Radiographic analysis showed 5 osteolytic lesions (8%) around well-fixed tibial and femoral components and minimal (1 mm) narrowing of the medial polyethylene thickness in 7 knees (12%). There were 6 (10%) failures requiring reoperation because of aseptic loosening (4 knees), deep infection (1 knee), and recurrent patellar dislocation (1 knee). Survivorship analysis using revision as the endpoint showed a cumulative success rate of 92% at 10 years. In this study, the IBPS knee has shown good long-term results with low rates of aseptic loosening and no failures attributable to polyethylene wear. PMID- 10475555 TI - Changes in knee alignment after total knee arthroplasty. AB - Changes in limb alignment after total knee arthroplasty were evaluated in 20 knees replaced with the Miller Galante knee system. The mean follow-up period was 87.4 months. Seventeen of the 20 knees were in the varus position on the initial postoperative radiographs, but the alignment significantly changed to become even more aligned toward varus during the follow-up period. The thickness of the ultra high-molecular-weight polyethylene (UHMWPE) also decreased significantly in the medial femorotibial joint. The wear of the UHMWPE possibly changed the alignment, and the postoperative alignment had a positive correlation with the wear rate. The components should be implanted so that the mechanical axis intersects the center of the components to prevent worsening of alignment as well as to minimize any such wear. PMID- 10475556 TI - Total hip arthroplasty in patients on long-term renal dialysis. AB - End-stage renal failure patients on long-term renal dialysis who underwent total hip arthroplasty (THA) were followed. Fifteen hips were implanted in 12 patients. There was a high mortality (58%) and high overall early complication rate (58%) with a deep infection rate of 13%. Of patients, 76% (n = 11) had good clinicoradiologic outcome of the THA before their death or at their latest follow up. THA in patients on dialysis, however, should be reserved for those among this group who are expected to have a better life expectancy. PMID- 10475557 TI - Patient-reported health status in total joint replacement. AB - A total of 176 consecutive patients undergoing either total hip or total knee arthroplasty were given SF-36 surveys at 4 time intervals: preoperatively and at 3 months, 6 months, and 1 year postoperatively. Transformed SF-36 scores were compared at the preoperative and 1-year time intervals between several demographic variables, including age, gender, number of sides replaced (unilateral vs bilateral procedure), and type of joint replacement surgery (knee or hip). Joint replacement surgery seemed to correct various self-reported health status differences that appeared preoperatively in demographic groups. PMID- 10475558 TI - 5- to 8-year results of the Freeman press-fit hip arthroplasty without HA coating: a clinicoradiologic study. AB - Sixty-one consecutive patients (68 hips) with an average age of 49.7 years (range, 21-72 years) had the Freeman uncemented hip arthroplasty without HA coating performed in the Southern General Hospital between 1987 and 1990. These patients were reviewed retrospectively for a mean duration of 66 months (range, 3 103 months). A high incidence of significant thigh pain (40.4%) at 5 to 8 years was noted in this series. The average 5- to 8-year femoral subsidence was 5.4 mm (range, 0-16.1 mm) in 45 hips. Seventeen hips (29.3%) required revision at the time of analysis. Nine hips (15.5%) were revised for aseptic loosening at an average duration of 54 months after the index operation. The conclusion drawn from this series is that the Freeman press-fit hip produced unsatisfactory medium term results. PMID- 10475559 TI - Gait affects tibial component fixation. AB - The prognosis of fixation in patients with total knee arthroplasty was classified as either good or poor based on migration results over 4 to 8 years using roentgen stereophotogrammetry. Gait analysis with the Vicon system was performed in 27 asymptomatic patients (1 bilateral), selected according to gender, implant design, age, and fixation principle to form two equivalent groups with different prognoses. The poor prognosis group walked with a predominantly flexing moment and higher moment peaks in the sagittal plane compared with the good prognosis group, in which moments were abnormally small. We conclude that individual gait patterns and subsequent differences in joint loading affect tibial component fixation. PMID- 10475560 TI - Autonomic neuropeptides in the interface membrane of aseptic loose hip prostheses. AB - We analyzed the presence of autonomic nerve fibers in the interface membranes (n = 9) surrounding aseptic loosened hip prostheses by immunohistochemistry. The study focused on the autonomic messengers neuropeptide Y (NPY), tyrosine hydroxylase (TH), the rate-limiting enzyme in the synthesis of noradrenaline (NA), and vasoactive intestinal polypeptide (VIP). Protein gene product (PGP) 9.5, a general marker of peripheral nerve fibers, was also analyzed to establish the neuronal character of the immunoreactive structures. PGP 9.5-positive and NPY positive nerve fibers were identified in all 9 samples, and VIP-immunoreactive and TH-immunoreactive fibers were found in 7. There was a difference in the distribution of nerve fibers both between and within the samples. Among the neuropeptides analyzed, NPY was most abundant. NPY-positive and TH-positive fibers were predominantly found around the blood vessel walls forming varicose nerve terminals. VIP-positive fibers were mainly observed as thin varicose nerve terminals with no relationship to blood vessels. Autonomic neuropeptides exert not only vasoactive and immunoregulatory effects, but also have been found to have direct effects on bone tissue. Moreover, the autonomic nervous system has been strongly implicated in nociception and inflammation. Neuronal NPY, TH, and VIP in the interface membrane may prove to contribute to the pathologic mechanisms leading to aseptic loosening of hip prostheses. PMID- 10475561 TI - Patellar contact forces with and without patellar resurfacing in total knee arthroplasty. AB - Patellar contact forces were measured in 7 fresh-frozen cadaver knees during continuous flexion-extension cycling for the normal knee and after total knee arthroplasty without and with patellar resurfacing using 2 prosthesis systems with different designs for the femoral and patellar components. Analysis of variance with specimen as a repeated measure was used to examine the effect of contact surface. When compared to normal knees, total knee arthroplasty without patellar resurfacing resulted in no change in patellar contact forces. The magnitude of the inferosuperior patellar shear force increased significantly after patellar replacement with an oval, biconcave patellar component and increased further after replacement with an oval, domed patellar component. There were no significant changes in the mediolateral and anteroposterior contact force components after patellar replacement. PMID- 10475562 TI - Torsional stability of primary total knee replacement tibial prostheses: a biomechanical study in cadaveric bone. AB - The significance of torsional loads on the tibial component of total knee replacement prostheses is not clearly established. This study investigated the micromotion induced by torsion, under physiologic loads. Three forms of fixation were sequentially tested in cadaveric tibiae: press-fit, press-fit plus 4 peripheral corticocancellous screws, and horizontally cemented. The implanted tibia was loaded with an axial force between 0.5 and 2.2 kN and a cyclic torque of +/- 5 Nm. The relative motion between the prosthesis and the bone was measured. Our results show little difference between the 3 methods of fixation at high axial loads. At low axial loads, however, the press-fit prosthesis may permit micromotions high enough to interfere (theoretically) with bone ingrowth. Motion is reduced when the same prosthesis is augmented with screws. The cemented prosthesis produces the lowest average micromotion. Our study indicates that rotational micromotion is unlikely to contribute to the failure of ingrowth into the uncemented tibial prosthesis under normal physiologic loads. At low axial loads, which may be encountered in the immediate postoperative period, ingrowth may be compromised. Thus, initial rotational stability is an important consideration in uncemented total knee replacement design and postoperative management. In poor-quality tibiae, cementation may be the only method of fixation to provide sufficient torsional stability. PMID- 10475563 TI - Plasticity-induced damage layer is a precursor to wear in radiation-cross-linked UHMWPE acetabular components for total hip replacement. Ultra-high-molecular weight polyethylene. AB - The mechanism for the improved wear resistance of cross-linked ultra-high molecular-weight polyethylene (UHMWPE) remains unclear. This study investigated the effect of cross-linking achieved by gamma irradiation in nitrogen on the tribologic, mechanical, and morphologic properties of UHMWPE. The goal of this study was to relate UHMWPE properties to the wear mechanism in acetabular-bearing inserts. Wear simulation of acetabular liners was followed by detailed characterization of the mechanical behavior and crystalline morphology at the articulating surface. The wear rate was determined to be directly related to the ductility, toughness, and strain-hardening behavior of the UHMWPE. The concept of a plasticity-induced damage layer is introduced to explain the near-surface orientation of the crystalline lamellae observed in the wear-tested acetabular liners. Cross-linking reduces abrasive wear of acetabular components by substantially reducing--but not eliminating--the plasticity-induced damage layer that precedes abrasive wear. PMID- 10475564 TI - The use of a flexible biodegradable cement restrictor in total hip arthroplasty. PMID- 10475565 TI - Inguinal abscess: an unusual presentation of infection around total hip replacement. AB - We describe 2 patients who presented with inguinal abscesses arising from infected total hip replacement. Recognition of the source of the infection was delayed. Both infections were eventually controlled after incision and drainage with removal of the infected prosthesis. PMID- 10475566 TI - Traumatic aneurysm of the inferior lateral geniculate artery after total knee replacement. AB - A 55-year old man incurred a false aneurysm of the inferior lateral geniculate artery after total knee replacement. The false aneurysm appeared as a pulsatile mass with compressive neuropathy of the posterior tibial nerve. The development of this complication and treatment are discussed. PMID- 10475567 TI - Severe polyethylene wear in uncemented acetabular cup system components: a report of 5 cases. AB - We report 5 cases of severe failure of the polyethylene liner at 4 to 7.4 years after uncemented anatomic medullary locking (AML) total hip arthroplasty, using the acetabular cup system (ACS). The mode of failure was by a combination of wear through to the metal backing and liner fracture. Four of the 5 cases were associated with a 32-mm head. The ACS design is flawed because of a lack of hemispherical geometry resulting in rim loading on a region of the liner that has the thinnest polyethylene. All patients with this type of implant should be monitored for evidence of excessive polyethylene wear. PMID- 10475568 TI - Factors affecting femoral bone remodeling after cementless total hip arthroplasty. AB - We performed a postmortem comparison of femurs from two patients who had bilateral cementless total hip arthroplasties with femoral prostheses of different stiffness implanted in their right and left hips. Radiographs of transverse sections of the four femurs demonstrated that all the prostheses were bone ingrown with the most ingrowth occurring distally where the porous coating contacted diaphyseal bone. In both patients, dual-energy x-ray absorptiometry analysis revealed that the femur implanted with the stiffer prosthesis had a 65% to 79% greater loss of proximal periprosthetic bone than the femur implanted with the more flexible prosthesis. One patient, however, had a dramatically greater total loss of bone from side to side than the other patient. In this patient, we believe that it was host factors more than the differences in stem stiffness that affected the bone-remodeling pattern. Although the two femurs with the stiffer prostheses had the greatest bone loss, the two femurs with the more flexible prostheses demonstrated radiographic signs of cantilever bending of the prosthetic stem and failure of proximal osseointegration. We are not aware of any other bilateral human postmortem analysis that so clearly illustrates the effect of stem stiffness on bone remodeling. PMID- 10475569 TI - NADPH-diaphorase and NOS-1 positive ganglion cells are found in the rat vallate papilla/von Ebner gland complex. AB - The nervous system of the vallata papilla and von Ebner glands was investigated in the rat tongue. Cells involved in the production of nitric oxide were identified by immunohistochemical detection of neuronal nitric oxide synthase type-1 and by cytochemical detection of NADPH-diaphorase. The analysis of serial sections showed that a ganglion composed of about 180-190 neuronal cells was present between the vallata papilla and von Ebner glands. These cells were positive for nitric oxide synthase type-1 and NADPH-diaphorase. From the ganglion, we observed nitrergic fibres running: (a) in the lamina propria of the receptor-free mucosa; (b) just below the gustatory epithelium; (c) in the von Ebner glands; and (d) around the vascular system of the vallata papilla. Our study suggests that the nitrergic ganglion cells may mediate interactions between chemoreceptorial systems in the vallata papilla and secretory cells in the von Ebner glands and that nitric oxide could be involved in the regulation of the blood supply to the vallata papilla and in the regulation of the von Ebner glands. PMID- 10475570 TI - Dystrophin and utrophin complexed with different associated proteins in cardiac Purkinje fibres. AB - Abnormal dystrophin expression is directly responsible for Duchenne and Becker muscular dystrophies. In skeletal muscle, dystrophin provides a link between the actin network and the extracellular matrix via the dystrophin-associated protein complex. In mature skeletal muscle, utrophin is a dystrophin-related protein localized mainly at the neuromuscular junction, with the same properties as dystrophin in terms of linking the protein complex. Utrophin could potentially overcome the absence of dystrophin in dystrophic skeletal muscles. In cardiac muscle, dystrophin and utrophin were both found to be present with a distinct subcellular distribution in Purkinje fibres, i.e. utrophin was limited to the cytoplasm, while dystrophin was located in the cytoplasmic membrane. In this study, we used this particular characteristic of cardiac Purkinje fibres and demonstrated that associated proteins of dystrophin and utrophin are different in this structure. We conclude, contrary to skeletal muscle, dystrophin-associated proteins do not form a complex in Purkinje fibres. In addition, we have indirect evidence of the presence of two different 400 kDa dystrophins in Purkinje fibres. PMID- 10475571 TI - Neutrophil gelatinase-associated lipocalin in normal and neoplastic human tissues. Cell type-specific pattern of expression. AB - Neutrophil gelatinase-associated lipocalin (NGAL) has recently been identified in myeloperoxidase-negative neutrophil granules. Members of the lipocalin family are thought to bind and transport small lipophilic molecules such as retinoids and roles in cell regulation have been proposed. Recently, NGAL has also been demonstrated in the colonic mucosa in certain pathologic conditions. The aim of this study was to examine the distribution of NGAL in normal and neoplastic tissues by immunohistochemistry. Interestingly, NGAL was found in a variety of normal and pathological human tissues. A cell type-specific pattern of expression was seen in bronchus, stomach, small intestine, pancreas, kidney, prostate gland, and thymus. The comparative analysis of the putative rat homologue neu-related lipocalin showed a very similar pattern of expression with the exception of pancreas and kidney. Neoplastic human tissues showed a very heterogeneous expression of NGAL protein. High NGAL levels were found in adenocarcinomas of lung, colon and pancreas. In contrast, renal cell carcinomas of various subtypes and prostate cancers contained low NGAL levels. Lymphomas and thymic tumours were negative for NGAL immuno-labeling. Knowledge about the location of NGAL in normal cells and in disease states provides the first clues towards understanding its biological function. PMID- 10475572 TI - Localization of nitric oxide-related substances in the quail ovary during folliculogenesis. AB - In the present study, nitric oxide (NO)-related substances, namely NO synthase (NOS), L-citrulline, cGMP and nitrotyrosine, have been localized in the quail ovary, using NADPH-diaphorase staining and immunohistochemical methods. The results indicate the presence of the NOS isoforms, showing distinct cell-specific distribution patterns in the quail ovary. Inducible NOS is primarily present in leukocytes, endothelial NOS in granulosa cells, and neuronal NOS in nerve cells, oocytes, interstitial cells and granulosa cells of pre-hierarchal follicles and of the germinal disc region of pre-ovulatory follicles. NOS activity, indicated by the presence of L-citrulline, is observed in oocytes, nerve cells, interstitial cells and a few granulosa cells of pre-hierarchal follicles. Detection of accumulated cGMP indicates that granulosa cells of pre-hierarchal and of pre- and post-ovulatory follicles, the theca interna of pre-ovulatory follicles, and oocytes are main targets of NO. Nitrotyrosine, a marker of peroxynitrite activity, is mainly localized in atretic follicles and in post ovulatory follicles. It is concluded that the quail ovary possesses a NO/NOS system, and that NO may be considered as a mediator involved in various ovarian processes, including atresia. PMID- 10475573 TI - Calcified microspheres as biological entities and their isolation from bone. AB - Calcified microspheres, about 1 microm in diameter, appear at sites of bone formation where they invest the collagenous matrix, become confluent and disappear. Evidence that the particle boundaries are not lost with compaction but merely deformed is supported in section by the granular histochemical staining of the inorganic phase for bone salt, lipid, fibronectin and acid phosphatase in osteomalacic, acid-etched and normal human bone. Their persistence as discrete objects is confirmed by the application of methods for their isolation from the collagenous matrix of immature mouse calvarium and mature bovine femur. Five methods have been used to extract them and include (i) biochemical, (ii) chemical, (iii) mechanical, (iv) pyrogenous and (v) biological separation. Under the optical microscope, all isolates consisted of similar discrete objects and bridged assemblies, whose birefringence varied with treatment. After decalcification, their organic 'ghosts' remained. Each isolated microsphere had a complex substructure of clusters of non-collagenous calcified filaments surrounding a less dense centre. The filaments were 5 nm in diameter with a 5 nm periodicity and regular fine interfilamentous connections. It is concluded that the microspheres are independent, complex, pervasive and central to the containment (i.e. packaging) of calcium phosphate in bone. Their extraction will enable further analysis. PMID- 10475574 TI - Thrombospondin is sequentially expressed and then de-expressed during early pregnancy in the rat uterus. AB - The expression of thrombospondin on Day 1, Day 3 and Day 6 of pregnancy has been examined in the rat, using light microscopic immunoperoxidase and electron immunogold techniques. The glycoprotein was expressed in the apical, lateral and basal uterine epithelium on Days 1 and 3 but was then de-expressed at the time of implantation on Day 6. We propose that these data suggest a role for thrombospondin in remodellig the uterine epithelium during the plasma membrane transformation, but that it does not play a part in attachment and implantation. PMID- 10475575 TI - Ultracytochemical study of guanylate cyclases A and B in light- and dark-adapted retinas. AB - The ultracytochemical localization of guanylate cyclases A and B activity has been studied after stimulation with atrial natriuretic peptide and C-type natriuretic peptide in light- and dark-adapted retinas and pigmented epithelium. The results showed that both peptides stimulated guanylate cyclases A and B activity in light-adapted retinas only. Guanylate cyclases A and B activity was detected on plasma membrane of body of photoreceptors, bipolar, horizontal and ganglion cells, on plasma membranes of interneuronal connections at plexiform layers and on the plasma membrane of fibres at the nerve fibres layer. Independently of the light-or dark-adapted state, the pigmented epithelium also presented guanylate cyclases A and B activity on basal and lateral plasma membranes. PMID- 10475576 TI - Binding properties of the galactose-detecting lectin Pseudomonas aeruginosa agglutinin (PA-IL) to skeletal muscle fibres. Quantitative precipitation and precipitation inhibition assays. AB - Pseudomonas aeruginosa agglutinin (PA-IL) staining and the influence of various carbohydrates on lectin binding to muscle sections were investigated quantitatively using a scanning and integrating microspectrophotometre. A strong dose-dependent inhibition of PA-IL staining in the sections was recorded with galabiose (Galalpha1-4Gal) while lactose (Galbeta1-4Glc) had no inhibitory effect. The affinity of PA-IL to Galalpha1 carbohydrates was studied by ELISA using immobilized glycoconjugates in which Galalpha1 glycans were attached to bovine serum albumin or ceramide. PA-IL exhibited strong binding to both simple glycoconjugates having a single Galalpha moiety and to di- and trisaccharides with terminal Galalpha1 at the non-reducing end. In all cross-sectioned muscle fibres incubated with PA-IL, the staining was present as a honeycomb-shaped network through the entire cytoplasm. Further, a dense punctuate staining could be shown in most fibres. A similar staining pattern was noticed after incubation with a monoclonal antibody against ryanodine receptors and with biotinylated ryanodine suggesting that the network could represent the sarcoplasmic reticulum. Further, Western blots of a sarcoplasmic reticulum preparation showed multiple bands after incubation with PA-IL. It may therefore be proposed that glycoconjugates carrying terminal Galalpha1 show affinity for PA-IL and are located to the sarcoplasmic reticulum. PMID- 10475578 TI - Optimizing electrode sites for segmental bioimpedance measurements. AB - Recent advances in the application of bioelectrical impedance analysis (BIA) have indicated that a more accurate approach to the estimation of total body water is to consider the impedance of the various body segments rather than simply that of the whole body. The segmental approach necessitates defining and locating the physical demarcation between both the trunk and leg and the trunk and arm. Despite the use of anatomical markers, these points of demarcation are difficult to locate with precision between subjects. There are also technical problems associated with the regional dispersion of the current distribution from one segment (cylinder) to another of different cross-sectional area. The concept of equipotentials in line with the proximal aspects of the upper (and lower) limbs along the contralateral limbs was investigated and, in particular, the utility of this concept in the measurement of segmental bioimpedance. The variation of measured segmental impedance using electrode sites along these equipotentials was less than 2.0% for all of the commonly used impedance parameters. This variation is approximately equal to that expected from biological variation over the measurement time. It is recommended that the electrode sites, for the measurement of segmental bioelectrical impedance in humans, described herein are adopted in accordance with the proposals of the NIH Technology Assessment Conference Statement. PMID- 10475577 TI - Characterization of common carotid artery blood-flow waveforms in normal human subjects. AB - Knowledge of human blood-flow waveforms is required for in vitro investigations and numerical modelling. Parameters of interest include: velocity and flow waveform shapes, inter- and intra-subject variability and frequency content. We characterized the blood-velocity waveforms in the left and right common carotid arteries (CCAs) of 17 normal volunteers (24 to 34 years), analysing 3560 cardiac cycles in total. Instantaneous peak-velocity (Vpeak) measurements were obtained using pulsed-Doppler ultrasound with simultaneous collection of ECG data. An archetypal Vpeak waveform was created using velocity and timing parameters at waveform feature points. We report the following timing (post-R-wave) and peak velocity parameters: cardiac interbeat interval (T(RR)) = 0.917 s (intra-subject standard deviation = +/- 0.045 s); cycle-averaged peak-velocity (V(CYC)) = 38.8 cm s(-1) (+/-1.5 cm s(-1)); maximum systolic Vpeak = 108.2 cm s(-1) (+/-3.8 cm s( 1)) at 0.152 s (+/-0.008 s); dicrotic notch Vpeak = 19.4 cm s(-1) (+/-2.9 cm s( 1)) at 0.398 s (+/-0.007 s). Frequency components below 12 Hz constituted 95% of the amplitude spectrum. Flow waveforms were computed from Vpeak by analytical solution of Womersley flow conditions (derived mean flow = 6.0 ml s(-1)). We propose that realistic, pseudo-random flow waveform sequences can be generated for experimental studies by varying, from cycle to cycle, only T(RR) and V(CYC) of a single archetypal waveform. PMID- 10475579 TI - The effects of motion on parametric fMRI analysis techniques. AB - Subject motion during the time course of functional activation studies has been shown to cause spurious signals which can mimic 'true' activation. Therefore, the importance of motion correction has been widely recognized. Correction with post processing using image registration software is common practice in functional imaging and analysis. Many image registration algorithms, developed for analysis requirements other than fMRI, assume rigid body motion. Although these techniques are now routinely used by a number of groups, rigid body coregistration has not yet been shown to reduce the effects of motion to an acceptable level in fMRI analysis, i.e. the effects on resulting correlation analysis directly. In this paper we have used volume data to assess rigid body co-registration in terms of motion artefacts for the different correlation approaches used in fMRI. We have developed a new way of visualizing motion effects in correlation analysis based on generating a scatter plot of correlation score against local image gradient. This technique has been tested on fMRI data sets from a functional paradigm suffering from motion correlated artefacts, with and without rigid body motion correction. Although we do not attempt to estimate the actual residual motion, this technique can be used to verify the results of analysis and select regions of relatively unambiguous activation. This paper assesses directly the rigid body assumption and proves the need for, and effectiveness of, co-registration for all correlation based analysis techniques. The specific differences between the popular correlation forms used are investigated and explained. We show that for certain forms of correlation analysis the effects of motion, while not removed altogether, are effectively statistically eliminated. PMID- 10475580 TI - Effects of CO2 on dynamic cerebral autoregulation measurement. AB - Arterial pCO2 is known to influence cerebral autoregulation but its effect on the dynamic relationship between mean arterial blood pressure (ABP) and mean cerebral blood flow velocity (CBFV), obtained from spontaneous fluctuations in ABP, has not been established. In 16 normal subjects, ABP was measured non-invasively (Finapres), CBFV was estimated with Doppler ultrasound in the middle cerebral artery, and end-tidal CO2 (EtCO2) was measured with an infrared capnograph. Recordings were made before, during and after breathing a mixture of 5% CO2 in air. The coherence function, amplitude and phase frequency responses, and impulse and step responses for the effects of ABP on CBFV were calculated by spectral analysis of beat-to-beat changes in mean ABP and CBFV before (mean CO2 5.55 +/- 0.38 kPa), during (6.43 +/- 0.31 kPa) and after 5% CO2 (5.43 +/- 0.26 kPa). During 5% CO2, the coherence function and the amplitude frequency response were significantly increased for frequencies below 0.05 Hz and the phase was reduced for the frequency range 0.02-0.1 Hz. The impulse and step responses indicated that 5% CO2 reduces the efficiency of the autoregulatory mechanism. A 20.7% average increase in CBFV induced by a 14.4% increase in EtCO2 was found to be mediated by a 25.9% reduction in critical closing pressure, while the change in resistance area product was non-significant. PMID- 10475581 TI - Latex vessels with customized compliance for use in arterial flow models. AB - The physical characteristics of the arterial wall exert a major influence over blood flow patterns and the pulse wave velocity is strongly affected by the elasticity of the vessel. We have developed a technique for manufacturing latex tubing which has physical characteristics similar to those of human arteries. Tubes were produced by painting a plastic rod of diameter 7 x 10(-3) m (the internal diameter of the superficial femoral artery) with liquid latex. The number of coats applied controlled the thickness, and hence the elasticity of the resulting tube. Values of compliance and pulse wave velocity were similar to those observed in vivo. The tubing was tested in a model of the femoral arterial circulation and the typical triphasic arterial now was seen. The mean flow in the vessels was also analysed, and found to be higher for the more elastic vessels, supporting existing mathematical theories and qualitative physiological data. These vessels provide a reliable method of producing physiologically accurate test segments for use in a range of arterial flow models. PMID- 10475582 TI - Modelling the relationship between peripheral blood pressure and blood volume pulses using linear and neural network system identification techniques. AB - The relationships between peripheral blood pressure and blood volume pulse waveforms can provide valuable physiological data about the peripheral vascular system, and are the subject of this study. Blood pressure and volume pulse waveforms were collected from 12 normal male subjects using non-invasive optical techniques, finger arterial blood pressure (BP, Finapres: Datex-Ohmeda) and photoelectric plethysmography (PPG) respectively, and captured to computer for three equal (1 min) measurement phases: baseline, hand raising and hand elevated. This simple physiological challenge was designed to induce a significant drop in peripheral blood pressure. A simple first order lag transfer function was chosen to study the relationship between blood pressure (system input) and blood volume pulse waveforms (system output), with parameters describing the dynamics (time constant, tau) and input-output gain (K). Tau and K were estimated for each subject using two different system identification techniques: a recursive parameter estimation algorithm which calculated tau and K from a linear auto regressive with exogenous variable (ARX) model, and an artificial neural network which was trained to learn the non-linear process input-output relationships and then derive a linearized ARX model of the system. The identification techniques allowed the relationship between the blood pressure and blood volume pulses to be described simply, with the neural network technique providing a better model fit overall (p < 0.05, Wilcoxon). The median falls in tau following the hand raise challenge were 26% and 31% for the linear and neural network based techniques respectively (both p < 0.05, Wilcoxon). This preliminary study has shown that the time constant and gain parameters obtained using these techniques can provide physiological data for the clinical assessment of the peripheral circulation. PMID- 10475583 TI - Microspectrophotometric analysis of respiratory pigments using a novel fibre optic dip probe in microsamples. AB - A microspectrophotometer system to monitor the reduction of mitochondrial respiratory pigments in cell extracts and permeabilized cells has been developed. The novel optical fibre set-up uses visible spectrophotometry to measure the reduction of mitochondrial electron carriers. The basis of the system is an Ocean Optics S1000 spectrometer, a broadband tungsten based light source, input and output coupling fibre optics and a fibre optic dip-probe which requires less than 20 microl of sample for analysis. The spectral range of the system is from 250 to 850 nm with a spectral resolution of 0.5 nm. Data are presented for the reduction of purified cytochrome c by the reducing agent sodium dithionite and the reduction of cytochrome c by isolated mitochondria using sodium succinate as substrate. Reduction of cytochrome c by digitonin permeabilized cultured mouse cells, C2C12, is also shown. The effect of temperature on cytochrome c reduction in these assays is also demonstrated. The optical design of the probe system is optimized to maintain maximum light throughput and spectral resolution. The key features of the system are small sample size, front-end adaptability, high sensitivity and fast multispectral acquisition which are essential for observing these biological reactions in vivo. PMID- 10475584 TI - Appropriate corrections to glomerular filtration rate and volume of distribution based on the bolus injection and single-compartment technique. AB - The bolus injection, single-compartment technique for measuring GFR overestimates the true value. Nevertheless, assuming that for a given indicator the area under the first exponential of the plasma clearance curve is constant from subject to subject, the observed (uncorrected) value can be corrected by multiplication with a 'sliding' factor, the value of which is a nonlinear function of GFR. Several second-order polynomials, based on pre-determined relationships between simultaneously determined two-compartment and one-compartment GFR, have been described for correcting GFR (GFR correction). It is, however, theoretically more rational to use a factor which depends on the rate constant, alpha2, of the terminal exponential of the clearance curve. We have therefore determined a set of linear equations from retrospectively analysed multiple-sample inulin, 99mTc DTPA and 51Cr-EDTA clearance curves to enable correction of GFR using alpha2. A set of linear equations is also developed to correct the volume of distribution (Vd) of the indicator (close to extracellular fluid volume for these indicators), which is also overestimated by the one-compartment technique. At low levels of GFR, alpha2-corrected GFR is similar to uncorrected GFR for all three indicators. As GFR increases, however, uncorrected GFR progressively overestimates (alpha2 corrected GFR. The overestimation is greater for inulin than for 99mTc-DTPA or 51Cr-EDTA. In the one-compartment approximation, Vd is overestimated more than GFR, and again the greatest overestimation is seen with inulin. In a prospective study of 129 patients undergoing routine measurement of GFR with 51Cr-EDTA, alpha2 correction using a factor based on retrospective EDTA data gave values of GFR which were higher than values obtained from GFR correction using a previously published polynomial (also based on EDTA clearances) by 15% in children and 12.5% in adults when uncorrected GFR was 150 ml/min/1.73 m2. Moreover, the ratio of uncorrected GFR to GFR-corrected GFR was higher in children than adults. We conclude that alpha2 is a more rational variable with which to correct two-sample or three-sample GFR than GFR itself, that the correction formulae are not interchangeable between inulin on the one hand and EDTA and DTPA on the other, and that the relative magnitudes of the corrections given by alpha2 correction versus GFR correction are different for children and adults. PMID- 10475585 TI - Fructose-2,6-bisphosphate and control of carbohydrate metabolism in eukaryotes. AB - Fructose-2,6-bisphosphate is an important intracellular biofactor in the control of carbohydrate metabolic fluxes in eukaryotes. It is generated from ATP and fructose-6-phosphate by 6-phosphofructo-2-kinase and degraded to fructose-6 phosphate and phosphate ion by fructose-2,6-bisphosphatase. In most organisms these enzymatic activities are contained in a single polypeptide. The reciprocal modulation of the kinase and bisphosphatase activities by post-translational modifications places the level of the biofactor under the control of extra cellular signals. In general, these signals are generated in response to changing nutritional states, therefore, fructose-2,6-bisphosphate plays a role in the adaptation of organisms, and the tissues within them, to changes in environmental and metabolic states. Although the specific mechanism of fructose-2,6 bisphosphate action varies between species and between tissues, most involve the allosteric activation of 6-phosphofructo-1-kinase and inhibition of fructose-1,6 bisphosphatase. These highly conserved enzymes regulate the fructose-6 phosphate/fructose-1,6-bisphosphate cycle, and thereby, determine the carbon flux. It is by reciprocal modulation of these activities that fructose-2,6 bisphosphate plays a fundamental role in eukaryotic carbohydrate metabolism. PMID- 10475586 TI - Manganese deficiency and toxicity: are high or low dietary amounts of manganese cause for concern? AB - Manganese is an essential trace element that is required for the activity of several enzymes. Manganese is also quite toxic when ingested in large amounts, such as the inhalation of Mn-laden dust by miners. This review examines Mn intake by way of the food supply and poses the question: Is there reason to be concerned with Mn toxicity or deficiency in free-living populations in North America? Although much remains to be learned of the functions of Mn, at present there are only a few vaguely described cases of Mn deficiency in the medical literature. Given the heterogeneity of the North American food supply, it is difficult to see the possibility of more than greatly isolated and unique instances of Mn deficiency. However, low Mn-dependent superoxide dismutase activity may be associated with cancer susceptibility, and deserves further study. There may be reasons, however, to be concerned about Mn toxicity under some very specialized conditions. Increasing numbers of young people are adopting a vegetarian lifestyle which may greatly increase Mn intake. Iron deficiency may increase Mn absorption and further increase the body-burden of Mn, especially in vegetarians. Mn is eliminated primarily through the bile, and hepatic dysfunction could depress Mn excretion and further contribute to the body burden. Would such a combination of events predispose substantial numbers of people to chronic Mn toxicity? At present, there is no definite proof of this occurring, but given the state of knowledge at the present time, more studies with longer time-frames and more sensitive methods of analysis are needed. PMID- 10475587 TI - Evidence for A1 and A2B adenosine receptors in baby hamster kidney (BHK) cells. AB - Adenosine is known to produce biphasic effects in the renal tissues via adenosine receptors. However, the presence of more than one subtype of adenosine receptor on a type of kidney cell or tissue has not been conclusively demonstrated. To address this issue, we investigated the presence of A1 and A2 adenosine receptors in baby hamster kidney (BHK) cells by use of radioligand binding and the reverse transcription-polymerase chain reaction. Ligand binding studies with (3H)-DPCPX revealed a single class of binding site with a K(D) of 9.2 +/- 2.0 nM, a Bmax of 1.7 +/- 0.2 pmol/mg protein and a pharmacological profile characteristic of A1 adenosine receptor on the BHK cell membrane. As the presence of A2 adenosine receptors could not be conclusively determined by ligand binding studies, the more sensitive method of RT-PCR was employed. The presence of A1 and A2B adenosine receptors was detected by RT-PCR with specific primers and the subsequent sequencing of the resultant amplification product. The sequences obtained were 75-90% homologous to the respective adenosine receptor mRNA of rat, mouse and human. PMID- 10475588 TI - Production and characterization of monoclonal antibodies to porcine brain pyridoxal kinase. AB - Six monoclonal antibodies that recognize porcine brain pyridoxal kinase have been selected and designated as PK67, PK86, PK91, PK144, PK252 and PK275. A total of six monoclonal antibodies recognizing different epitopes of the enzyme were obtained, of which four inhibited the enzyme activity. When total proteins of porcine brain homogenate separated by SDS-PAGE were subjected to monoclonal antibodies, a single reactive protein band of molecular weight 39 kDa which comigrated with purified porcine pyridoxal kinase was detected. Using the anti pyridoxal kinase antibodies as probes, the cross reactivities of brain pyridoxal kinase from human and other mammalian tissues and from avian sources were also investigated. Among human and all animal tissues tested, immunoreactive bands on Western blots appeared to have the same molecular mass of 39 kDa. These results indicate that mammalian brains contain only one major type of immunologically similar pyridoxal kinase, although some properties of the enzymes reported previously differed from one another. PMID- 10475589 TI - Vanadium and diabetes. AB - Vanadium is an ultratrace element, widely distributed in nature, yet with no presently known specific physiological function in mammals. The apparent role of vanadium in regulation of intracellular signaling, as a cofactor of enzymes essential in energy metabolism, and as a possible therapeutic agent in diabetes is of increasing interest as more and more research reports present evidence of vanadium's potentially unique biological function. In this mini-review, the author summarizes current knowledge of the bioinorganic chemistry of vanadium, the basic features of diabetes mellitus and its metabolic sequelae, and the in vitro and in vivo effects of both inorganic and organically-chelated vanadium compounds. Results of clinical trials to date, as well as kinetic studies of tissue uptake are covered. Examples of ways to enhance the positive effects of vanadium as an oral therapeutic adjunct in diabetic control, while minimizing potential toxicity, are compared with regard to desirable features and possible drawbacks. PMID- 10475590 TI - Copper and signal transduction: platelets as a model to determine the role of copper in stimulus-response coupling. AB - Platelets from copper-deficient rats have been used as a model to investigate the role of copper in receptor-mediated cellular responses. Copper deficiency doubles the rate of dense granule secretion and increases myosin association with the platelet cytoskeleton following thrombin stimulation. Mechanisms underlying the effects of copper deficiency on thrombin-induced signals that elicit dense granule secretion involve suppression of protein kinase C activity and impairment of Ca2+ release from intracellular stores. Copper deficiency also reduces the cellular GTP content of platelets. This may limit receptor effector coupling through GTP-dependent regulatory proteins leading to protein kinase C activation and the release of Ca2+ from intracellular stores. The reduction in GTP content during copper deficiency results from its utilization to maintain cellular ATP levels in response to severely inhibited cytochrome c oxidase activity in platelet mitochondria. Thus, the role of copper in maintaining normal signal transduction may be indirectly related to its biological function in mitochondria. PMID- 10475591 TI - Captopril increased mitochondrial coenzyme Q10 level, improved respiratory chain function and energy production in the left ventricle in rabbits with smoke mitochondrial cardiomyopathy. AB - The aim of the study was to show whether the ACE inhibitor captopril is able to protect the heart against the deleterious effect of passive cigarette smoking on left ventricular mitochondria. Four groups of rabbits were investigated: control (C), passive smoking of three cigarettes twice daily/30 minutes (S), control + captopril (7.5 mg/kg body weight twice daily) (Cap), and smoking + captopril (SCap) as in group 2 and 3. Three weeks lasting passive smoking impaired oxidative phosphorylation, diminished cytochrome oxidase activity and increased the mitochondrial F1-ATPase protein concentration. Moreover, the level of coenzyme Q10 (CoQ10) and coenzyme Q9 were decreased. Simultaneous treatment with captopril prevented partly the decrease of CoQ10 level, deterioration of oxidative phosphorylation, diminution of cytochrome oxidase activity and enhancement of F1-ATPase level. We conclude that captopril protected the myocardium against the harmful effect of passive smoking in rabbits. PMID- 10475592 TI - Influence of dietary nucleotides on plasma immunoglobulin levels and lymphocyte subsets of preterm infants. AB - We examined the effects of nucleotide supplementation to a preterm adapted milk formula on the lymphocyte subsets and plasma IgG, IgM and IgA levels in preterm infants for the first three months of life. Two groups of preterm infants received a milk formula or the same formula supplemented with CMP, AMP, UMP, GMP and IMP to mimic the concentration of acid-soluble nucleotides found in human milk. Blood samples were obtained at birth, 10 days, 20-30 days and 3 months of age. Preterm infants fed the nucleotide formula exhibited higher plasma levels of IgM in all postnatal study periods than neonates fed the standard formula; moreover, IgA was also higher at 3 months of age in nucleotide formula fed infants. No major differences were seen between groups for IgG levels and lymphocyte subsets. Thus, dietary nucleotides appear to exert actions on immature human neonate lymphocytes enhancing the in vivo production of Ig which may have a role in the defense capacity of neonates. PMID- 10475593 TI - Pediatric pain: new directions from a developmental perspective. PMID- 10475594 TI - Pediatric fibromyalgia and dizziness: evaluation of vestibular function. AB - Twelve children with fibromyalgia and complaints of chronic dizziness were evaluated with both clinical office maneuvers of vestibular function and laboratory tests composed of electronystagmography and sinusoidal harmonic acceleration rotary chair testing. All test results were normal for spontaneous nystagmus with or without visual fixation, oculocephalic reflex, dynamic visual acuity, head-shaking nystagmus, Quix test, and Dix-Hallpike maneuver. Electronystagmography test results were essentially normal for saccades, gaze, Dix-Hallpike, pendular tracking, and caloric evaluation. Rotary chair testing was normal in all 12 patients. These findings suggest that central (brainstem) and peripheral vestibular (inner ear) mechanisms do not account for the complaints of dizziness in the pediatric patient with fibromyalgia. The common musculoskeletal abnormalities of fibromyalgia may affect their proprioceptive orientation, therefore giving them a sense of imbalance. PMID- 10475595 TI - Management of pain from heel lance with lidocaine-prilocaine (EMLA) cream: is it safe and efficacious in preterm infants? AB - Hospitalized preterm infants undergo multiple painful heel lances. A two-phase, randomized, controlled trial was undertaken to determine the safety and efficacy of lidocaine-prilocaine 5% cream (EMLA, Astra Pharmaceuticals, L.P, Westborough, MA) for relieving pain from heel lance. One hundred twenty infants were randomly assigned to receive 0.5 g of EMLA or placebo cream for 30 minutes (Phase 1) or 60 minutes (Phase 2) before a routine heel lance. Efficacy was assessed using the Premature Infant Pain Profile (PIPP). Safety was determined by methemoglobin concentration 8 hours after EMLA application and by clinical signs of methemoglobinemia. No significant differences existed on PIPP scores between EMLA and placebo groups in Phase 1 (p < .480) or Phase 2 (p < .831). No infant had any clinical signs of methemoglobinemia. The mean methemoglobin concentration was 1.19% (.47). Approximately 10% of infants had minor skin reactions, and approximately 20% of EMLA-treated infants had blanching at the application site. The authors conclude that EMLA is safe but not efficacious for relieving pain from heel lance in preterm infants. PMID- 10475596 TI - Clinimetric evaluation of the pain observation scale for young children in children aged between 1 and 4 years after ear, nose, and throat surgery. AB - This study assessed the reliability, validity, and responsiveness of a new pain measure for children aged 1 to 4 years that was developed from the Children's Hospital of Ontario Pain Scale and its Neonatal Infant Pain Scale. Pain in 311 children, aged 1 to 4 years, was measured by two observers at fixed intervals after adenotonsillectomy (n = 114), adenotomy (n = 109), or insertion of ventilation tubes (grommets) (n = 88) until discharge using a dichotomous pain scale of 9 behavioral and physiological categories. The scale proved to be strongly homogeneous. The interobserver agreement was substantial for 7 items. On these final 7 items, the ability to distinguish between patients with differing degrees of pain and the sensitivity to detect changes over time within each patient were substantial. The resulting Pain Observation Scale for Young Children is reliable and easy to use for assessment of short- and longer-lasting pain after ear, nose, and throat surgery and may be used for assessing pain with other conditions. PMID- 10475597 TI - Children with cognitive impairment: parent report of pain and coping. AB - Information about pain in children with cognitive impairment is lacking. To gather pain-relevant information in this population, parents of 145 children with borderline to profound cognitive impairment were interviewed regarding their children's pain expression, experience, treatment, and coping behavior. Descriptions of pain expression and coping behavior were associated with the level of cognitive impairment. Children with mild to moderate cognitive impairment were more likely to be described as directly communicating their pain and exhibiting procedural coping strategies similar to those observed in children without cognitive impairments. More than half of the parents reported that their children experienced pain differently than did children without cognitive impairment, with the majority perceiving decreased pain sensitivity and greater pain tolerance. Finally, one third of parents felt that their children's pain was treated differently than that of other children. Half of these parents believed that health care providers had difficulties assessing and treating their children's pain. Results contribute to the developing foundation of information about pain in this special pediatric population. PMID- 10475598 TI - Pain in children with significant neurological impairment. AB - The pain experience in the child with a significant neurological impairment is complex and confusing, and it raises many questions about the very nature of pain itself. Early work in this field suggests that the pain experience may be blunted. The neurological impairment associated with conditions such as cerebral palsy may alter the neurological system and hence the ability to comprehend and communicate pain; there is no evidence to date that this reflects true pain insensitivity or indifference. From recent work, the emerging body of evidence supports a relationship between the pain system and the motor, sensory, and autonomic systems and demonstrates how alterations to these systems may have a profound and unique impact on the pain experience. Beyond the altered neurological substrate, communication disabilities and social/environmental factors also seem to alter the pain experience. Establishing a clear pain history, including baseline information of child-specific patterns of behaviors and ongoing comparative use of this information over time, can provide clinically meaningful measures. Pain management should be directed at the underlying sources of pain and should include the analgesic ladder for everyday pain, opioids for acute/procedural pain (+/- benzodiazepine adjuvants), and antispasticity medications for high tone. With appropriate monitoring, demand and regional analgesic techniques can provide effective and safe postoperative pain control. The lack of basic and clinical knowledge in this field, however, adds another challenge to the clinician. PMID- 10475599 TI - Importance of sleep in the management of pediatric pain. AB - This article outlines several aspects of sleep regulation relevant to pediatric pain management. A broad range of connections between sleep and pain are described: (1) pain can interfere with the quality and quantity of children's sleep; (2) insufficient sleep (quality or quantity) can cause daytime sequelae (behavioral and emotional changes) that interfere with the coping skills necessary for effective pain management; (3) fear and anxiety often have a negative impact on both pain and sleep; (4) feelings of safety and control frequently have a positive effect on both sleep and pain symptoms; (5) adequate sleep seems to promote both physiological (tissue repair) and psychological (transient cessation of the perception of pain signals) processes relevant to recovery from pain, injury, and illness; and (6) treatment approaches to pediatric sleep and pain problems show considerable overlap with respect to many pharmacological as well as cognitive-behavioral interventions. Given these multiple links, a better understanding of sleep--and its importance in physical and mental health--is likely to be of value to clinicians and researchers working in areas of pediatric pain management. One specific hypothesis to be addressed is the possible contribution of sleep disruption as a step in the progression to some chronic pain syndromes. PMID- 10475600 TI - Long-term effects of pain in infants. AB - Pain and stress have been shown to induce significant physiological and behavioral reactions in newborn infants, even in those born prematurely. Infants who are born prematurely or seriously ill are commonly exposed to multiple painful and stressful events as part of their prolonged hospitalizations and required medical procedures. There is now evidence that these early events not only induce acute changes, but that permanent structural and functional changes may also result. This article reviews the growing body of evidence of likely long term effects of early pain and stress on the human infant. It is hoped that a better understanding of this literature will promote more responsive and sensitive management of infants and young children during their encounters with the medical community and will ultimately facilitate the healthy growth and development of all children. PMID- 10475601 TI - Children's memory for pain. AB - Although there is a rich body of research on the development of children's memory for the details of personally experienced events, relatively little is known about age-related changes in the ability to remember pain. This gap in the literature is surprising, given that studies of children's memory for painful experiences are relevant to our basic understanding of cognitive development, pain perception, and--in some situations--patient management. This article examines what is known about children's memory for pain, given its inherent importance, working from the vantage point of the literature on the development of autobiographical memory. In doing so, the authors make use of an informal information-processing framework to organize their thoughts about the acquisition, retention, and distortion of information about painful experiences. Nonetheless, the authors recognize that this framework will no doubt need to be modified to take into account the complex memory representations--containing somatosensory, affective, and contextual information--that are established after exposure to painful stimulation. After the treatment of the literature, the authors discuss its implications for the clinical management of pain in pediatric settings. PMID- 10475602 TI - Review of juvenile primary fibromyalgia and chronic fatigue syndrome. AB - This article reviews the current literature on childhood fibromyalgia and chronic fatigue syndrome. In doing so, it questions assumptions about the presumed nature of the disorders-that they are distinct from each other and are duplicates of their adult counterparts. It also attempts to synthesize the available data to reach some preliminary judgments about these disorders: that fibromyalgia and chronic fatigue syndrome may be related in children and may not be duplicates of the adult disorders; that psychological and psychosocial factors are unlikely contributors to the etiology of these disorders; and that the evidence is increasingly pointing to a role for genetic factors in their etiology. A discussion of the research into treatments for childhood fibromyalgia and chronic fatigue syndrome highlights the lack of well-designed, controlled studies. Finally, directions for future research are offered where results of the current literature are unclear. PMID- 10475603 TI - "By the way,...her foot is turned out.". PMID- 10475604 TI - Overexpression of the Helix-Loop-Helix protein Id2 blocks T cell development at multiple stages. AB - The Id proteins are inhibitors of basic-Helix-Loop-Helix transcription factor function that have been implicated in the control of cell differentiation and proliferation. To study the role of Id proteins in the control of T cell development, we generated transgenic mice that overexpress the Id2 protein in thymocytes. We detect a significant expansion of the early CD4(-)CD8(+)TCR(-) thymocyte stage and a depletion of the thymocytes of the subsequent developmental stages. These data indicate that the overexpression of Id2 leads to a stage specific developmental block early in thymopoiesis. In addition, progeny mice from five of the six Id2 transgenic founder lines succumb to aggressive T cell hyperproliferation that resembles lymphoma. Thus, overexpression of the Id2 protein has profound effects on T cell development and oncogenesis, consistent with the hypothesis that the bHLH proteins play critical roles in these processes. PMID- 10475605 TI - Gene structure of the P100 serine-protease component of the human Ra-reactive factor. AB - The Ra-reactive factor (RaRF) is a complement dependent anti-microbial factor that reacts with numerous microorganisms such as viruses, bacteria, fungi and protozoa. It is a complex of a mannan-binding lectin (MBL) and the serine protease, P100 (MASPI). P100 activates the C4 component of the complement system and its domain organization is similar to C1r and C1s. In this study, determination was made of the structure of the human P100 gene which was found longer than 67 kbp and to be comprised of 16 exons. Its non-protease region consisted of 10 exons, as in the case of C1r and C1s, and the introns were found present in the boundary separating two CUB domains, an EGF-like domain and two CCP domains and each CUB and CCP domain contained extra internal introns. The serine protease region was comprised of 6 exons in contrast to C1r and C1s, either of which consists of a single exon. The exon-intron structure was found to reflect the evolution of these molecules and P100 to have derived earlier in the stage of evolution than C1r or C1s. PMID- 10475606 TI - Loss of splice consensus signal is responsible for the removal of the entire C(H)1 domain of the functional camel IGG2A heavy-chain antibodies. AB - The molecular basis for the absence of the C(H)1 domain in naturally occurring heavy-chain antibodies of the camelids was assessed by determining the entire Camelus dromedarius gamma2a heavy-chain constant gene. The organization of the camel gamma2a constant heavy-chain gene obtained from a liver genomic library appears to be typical of all other mammalian gamma genes sequenced to date. It contains the switch, CH1, hinge, CH2, CH3, M1 and M2 exons. In contrast to the case in mouse and human heavy chain diseases, the camel gamma2a gene shows no major structural defect, and its equivalent CHI exon is intact. However, sequence analysis has revealed that the splicing site, immediately after the CH1 exon, is defective due to point mutations, especially the G(+1) to A(+1) transversion seems to be detrimental. It is concluded that the loss of the splice consensus signal is responsible for the removal of the entire CH1 domain in camel gamma2a heavy-chain immunoglobulins. Additionally, a closer analysis of the hinge exon suggests the possible involvement of transposons in the genetic variation of mammalian Cgamma hinges. PMID- 10475607 TI - Immunosuppressory mini-regions of HLA-DP and HLA-DR. AB - Our previous studies showed, that the,TPQRGDVYT, QRGDVYT and RGDVYT fragments, located in the beta164-172 loop of HLA-DQ, strongly suppress the humoral and cellular immune response, while their shorter analogs, RGDV, RGDVY, and QRGDVY, show only weak stimulatory activity in respect to humoral immunological response. The fragments contain the RGDVY sequence that is analogous to thymopentin (pentapeptide RKDVY, an immune system activator) as well as the RGD sequence, known for its importance for cellular association phenomena. Based on the crystal structure of HLA-DR1, we also designed and synthesized a cyclic analog C*RGDVYC* (where C* indicates Cys participating in disulfide bridge) with restricted conformation, which strongly suppresses both humoral and cellular immune response. In the present study we synthesized and tested the immunological properties of the linear and cyclic HLA-DP and HLA-DR counterparts of all the above HLA-DQ fragments. Although the results show that the linear HLA-DP fragments possess moderate immunosuppressory potency, their conformationally restricted analog, C*QGDVYC*C shows a considerable suppression of both humoral and cellular immune response. The nonapeptide fragment of HLA-DR, VPRSGEVYT and particularly its cyclic analog C*SGEVYC*, are strong suppressors of the humoral response. PMID- 10475608 TI - Isolation and sequence of a cDNA coding for the heavy chain constant region of IgG from the Australian brushtail possum, Trichosurus vulpecula. AB - A brushtail possum (Trichosurus vulpecula) mesenteric lymph node cDNA library was screened with a South American short-tailed opossum (Monodlelphis domestica) immunoglobulin gamma heavy chain constant region (Cgamma) probe, resulting in the isolation of a 1518 nucleotide cDNA clone. The sequence corresponds to exons 1-3 of Cgamma. The Australian marsupial (T. vulpeculla) sequence is 70% identical at the amino acid level with the American marsupial (M. domestica) sequence, but less similar to the eutherian mammals (45-50%). These data provide the opportunity to compare the evolution of IgG between orders of marsupials separated by at least 75 million years and confirm the appearance of IgG prior to the metatherian/eutherian divergence. PMID- 10475609 TI - Calcium-induced ERK activation in human T lymphocytes. AB - We have previously shown that stimulation of B lymphocytes with calcium ionophores lead to the phosphorylation and enzymatic activation of ERK2. We have now determined that stimulation of human primary and Jurkat T lymphocytes with ionomycin also results in the activation of ERK1 and 2 as determined by; shifts in the mobility of this enzyme on SDS PAGE gels, the binding of an antibody that recognizes only the activated form of this enzyme, and increased ability to phosphorylate myelin basic protein (MBP). Another calcium ionophore, A23187, also induced activation of ERK1 and 2 in human primary and Jurkat T lymphocytes demonstrating that this is a general effect of calcium ionophores and is not limited to ionomycin. The activation of ERK1 and 2 by calcium ionophores was rapid, transient, and occurred in a dose-dependent manner. Activation of ERK1 and 2 by increases in intracellular calcium were blocked by the MEK inhibitor PD98059. These data point to a new role for calcium fluxes in T lymphocytes. PMID- 10475610 TI - Signal joint formation is inhibited in murine scid preB cells and fibroblasts in substrates with homopolymeric coding ends. AB - During B and T lymphocyte development, immunoglobulin and T cell receptor genes are assembled from the germline V, (D) and J gene segments (Lewis, S.M., 1994. The mechanism of V(D)J joining: lessons from molecular, immunological and comparative analyses. Adv. Immunol. 56, 27-150). These DNA rearrangements, responsible for immune system diversity, are mediated by a site specific recombination machinery via recognition signal sequences (RSSs) composed of conserved heptamers and nonamers separated by spacers of 12 or 23 nucleotides (Lewis, S.M., 1994. The mechanism of V(D)J joining: lessons from molecular, immunological and comparative analyses. Adv. Immunol. 56, 27-150). Recombination occurs only between a RSS with a 12mer spacer and a RSS with a 23mer spacer (Lewis, S.M., 1994. The mechanism of V(D)J joining: lessons from molecular, immunological and comparative analyses. Adv. Immunol. 56, 27-150). RAG1 and RAG2 proteins cleave precisely at the RSS-coding sequence border leading to flush signal ends and coding ends with a hairpin structure (Eastman, M., Leu, T., Schatz, D., 1996. Initiation of V(D)J recombination in vitro obeying the 12/23 rule. Nature 380, 85-88; Roth, D.B., Menetski, J.P., Nakajima, P.B., Bosma, M.J., Gellert, M., 1992. V(D)J recombination: broken DNA molecules with covalently sealed (hairpin) coding ends in scid mouse thymocytes. Cell 983-991: Roth, D.B., Zhu, C., Gellert. M., 1993. Characterization of broken DNA molecules associated with V(D)J recombination. Proc. Natl. Acad. Sci. USA 90, 10,788-10,792; van Gent, D., McBlane, J.. Sadofsky, M., Hesse, J., Gellert, M., 1995. Initiation of V(D)J recombination in a cell-free system. Cell 81, 925-934). Signal ends join, forming a signal joint. The hairpin coding ends are opened by a yet unknown endonuclease, and are further processed to form the coding joint (Lewis, S.M., 1994. The mechanism of V(D)J joining: lessons from molecular, immunological and comparative analyses. Ad. Immunol. 56, 27-150.) The murine scid mutation has been shown to affect coding joints, but much less signal joint formation. In this study we demonstrate that the murine scid mutation inhibits correct signal joint formation when both coding ends contain homopolymeric sequences. We suggest that this finding may be due to the function of the SCID protein as an assembly component in V(D)J recombination. PMID- 10475611 TI - Selenium effects on gallium arsenide induced biochemical and immunotoxicological changes in rats. AB - The influence of selenium (6.3 and 12.6 micromol/kg, intraperitoneally) on the disposition of gallium and arsenic and a few gallium arsenide (GaAs) sensitive biochemical variables was studied in male rats. Concomitant administration of Se and GaAs (70 micromol/kg, orally, 5 days a week for 4 weeks) significantly prevented the accumulation of arsenic while, the gallium concentration reduced moderately in the soft organs. The biochemical (haematopoietic and liver) and immunological variables however, responded less favorably to selenium administration. Most of the protection was however observed with the dose of 12.6 micromol rather than at 6.3 micromol. The results thus suggest a few beneficial effects of selenium in preventing the appearance of signs of GaAs toxicity like preventing inhibition of blood delta-aminolevulinic acid dehydratase (ALAD), hepatic malondialdehyde (MDA) formation and the accumulation of gallium and arsenic concentration. PMID- 10475612 TI - Oxygen activation during peroxidase catalysed metabolism of flavones or flavanones. AB - Flavonoids containing phenol B rings, e.g. naringenin, naringin, hesperetin and apigenin, formed prooxidant metabolites that oxidised NADH upon oxidation by peroxidase/H2O2. Extensive oxygen uptake occurred which was proportional to the NADH oxidised and was increased up to twofold by superoxide dismutase. Only catalytic amounts of flavonoids and H2O2 were required indicating a redox cycling mechanism that activates oxygen and generates H2O2. NADH also prevented the oxidative destruction of flavonoids by peroxidase/H2O2 until the NADH was depleted. These results suggest that prooxidant phenoxyl radicals formed by these flavonoids cooxidise NADH to form NAD radicals which then activated oxygen. Similar oxygen activation mechanisms by other phenoxyl radicals have been implicated in the initiation of atherosclerosis and carcinogenesis by xenobiotic phenolic metabolites. This is the first time that a group of flavonoids have been identified as prooxidants independent of transition metal catalysed autoxidation reactions. PMID- 10475613 TI - In vitro metabolism of alachlor by human liver microsomes and human cytochrome P450 isoforms. AB - Alachlor (2-chloro-N-methoxymethyl-N-(2,6-diethylphenyl)acetamide) is a widely used pre-emergent chloroacetanilide herbicide which has been classified by the USEPA as a probable human carcinogen. The putative carcinogenic metabolite, 2,6 diethylbenzoquinone imine (DEBQI), is formed through a complex series of oxidative and non-oxidative steps which have been characterized in rats, mice, and monkeys but not in humans. A key metabolite leading to the formation of DEBQI is 2-chloro-N-(2,6-diethylphenyl)acetamide (CDEPA). This study demonstrates that male human liver microsomes are able to metabolize alachlor to CDEPA. The rate of CDEPA formation for human liver microsomes (0.0031 +/- 0.0007 nmol/min per mg) is significantly less than the rates of CDEPA formation for rat liver microsomes (0.0353+/-0.0036 nmol/min per mg) or mouse liver microsomes (0.0106 +/- 0.0007). Further, we have screened human cytochrome P450 isoforms 1A1, 1A2, 2B6, 2C8, 2C9, 2C18, 2C19, 2D6, 2E1, and 3A4 and determined that human CYP 3A4 is responsible for metabolism of alachlor to CDEPA. Further work is necessary to determine the extent to which humans are able to metabolize CDEPA through subsequent metabolic steps leading to the formation of DEBQI. PMID- 10475614 TI - Investigation of benzene oxide in bone marrow and other tissues of F344 rats following metabolism of benzene in vitro and in vivo. AB - This study examines the initial activation of benzene, exploring key aspects of its metabolism by measurement of benzene oxide (BO) and BO-protein adducts in vitro and in vivo. To assess the potential influence of various factors on the production of BO, microsomes were prepared from tissues that were either targets of benzene toxicity, i.e. the bone marrow and Zymbal glands, or not targets, i.e. liver and kidneys, of control and acetone-treated F344 rats. No BO or phenol was detected in microsomal preparations of bone marrow or Zymbal glands (less than 0.007 nmol BO/mg protein and 0.7 nmol phenol/mg protein). On the other hand, BO and phenol were readily detected in preparations of liver and kidney microsomes and acetone pretreatment resulted in a 2-fold (kidney) increase or 3.7-fold (liver) increase in production of these metabolites. Initial rates of BO production in the liver isolates were 30 (control) to 50 (acetone-treated) times higher than in the corresponding kidney tissues. The estimated half-life of BO in bone marrow homogenates was 6.0 min and the second-order reaction rate constant was estimated to be 1.35 x 10(-3) l (g bone marrow)(-1) (h)(-1). These kinetic constants were used with measurements of BO-bone marrow adducts in F344 rats, receiving a single gavage dosage of 50-400 mg benzene (kg body weight)(-1) (McDonald, T.M., et al. (1994), Cancer Res. 54, 4907-4914), to predict the bone marrow dose of BO. Among the rats receiving 400 mg (kg body weight) (-1), a BO dose of 1.13 x 10(3) nM BO-h was estimated for the bone marrow, or roughly 40% of the corresponding blood dose predicted from BO-albumin adducts. Together these data suggest that, although BO is not produced at detectable levels in the bone marrow or Zymbal glands of F344 rats, BO is rapidly distributed via the bloodstream to these tissues where it may play a role in toxicity. PMID- 10475615 TI - Cellular thiols status and cell death in the effect of green tea polyphenols in Ehrlich ascites tumor cells. AB - Epidemiological studies suggest that the consumption of green tea may help prevent cancers in humans, and also breast and prostate cancers in animal models are reduced by green tea, and several mechanisms have been proposed for these effects. In this study the relationship between cellular sulfhydryl (SH) groups and the cytotoxicity of green tea polyphenols in Ehrlich ascites tumor cells was examined. It was found that in the presence of green tea extract (GTE) (100 microg/ml) and one of its polyphenolic components, epigallocatechin (EGC; 100 microM), both cellular non-protein (GSH) and protein-sulfhydryl (PSH) levels were significantly decreased and this was associated with a decrease in cell viability. Replenishing the thiol levels by using N-acetylcysteine (NAC) caused a recovery in cell viability, but this recovery was dependent on the time of thiol replenishment in the presence of EGC (initial 15 min). These results identify SH groups as a novel target of green tea polyphenols cytotoxicity in tumor cells, and a regulatory role for green tea in terms of reducing sulfhydryls in tumor inhibition. PMID- 10475616 TI - Apoptosis and its significance in MDS: controversies revisited. PMID- 10475617 TI - A phase III comparison of high dose ARA-C (HIDAC) versus HIDAC plus mitoxantrone in the treatment of first relapsed or refractory acute myeloid leukemia Southwest Oncology Group Study. AB - The aim of this study is to determine whether the addition of mitoxantrone to high dose cytarabine improves the outcome of treatment in patients with relapsed or refractory acute myeloid leukemia (AML). One hundred and sixty-two eligible patients, 14-76 years of age, with AML either in first relapse or that failed to respond to initial remission induction therapy, with no CNS involvement were randomized to receive therapy with cytarabine 3 gm/M2 i.v. over 2 h every 12 h for 12 doses on days 1-6 (Arm I) (HIDAC); or HIDAC plus mitoxantrone 10 mg/M2 i.v. daily on days 7 9 (Arm II) (HIDAC + M). Patients achieving complete remission were treated with three courses of consolidation including HIDAC (Ara-C 3 gm/M2 i.v. 12 h days 1 3; 2 gm/M2 over age 50) alone (ARM I) or with mitoxantrone (10 mg/M2 i.v. day 1) (ARM II). Among 162 patients (81 HIDAC, 81 HIDAC + M) evaluated for induction toxicity, there were 10 (12%) induction deaths with HIDAC and 13 (17%) with HIDAC + M (2-tailed P = 0.65). Most early deaths were due to infection and/or hemorrhage. Among 162 patients evaluated for responses to induction therapy, 26/81 (32%) HIDAC and 36/81 (44%) HIDAC + M patients achieved complete remission (two-tailed P = 0.15). Although this difference was not statistically significant in univariate analysis, it was after adjusting for the effects of WBC and PMN percentage in multivariate analysis (P=0.013). Median survivals from study entry were 8 months (HIDAC) and 6 months (HIDAC + M); 2-tailed logrank P = 0.58. Among 48 patients registered for consolidation, the median disease-free survivals from that registration were 8 months with HIDAC and 11 months with HIDAC + M (P = 0.60). There were three treatment-related deaths during consolidation (1 HIDAC, 2 HIDAC + M), all due to infections. In this randomized trial, the addition of mitoxantrone to high-dose cytarabine was associated with a trend toward a higher CR rate. There was less evidence for an advantage in disease-free or overall survival, although any such conclusion is limited by the size of the study. PMID- 10475618 TI - The prognostic significance of 13q14 deletions in chronic lymphocytic leukemia. AB - Although chronic lymphocytic leukemia (CLL) is the most common leukemia in adults, little is known about the molecular abnormalities underlying it and their prognostic significance. Using a battery of six microsatellite markers from 13q12.3-14.3 between BRACA2 gene and the Rb gene, we assayed loss of heterozygosity (LOH) in 78 CLL patients. We found deletion in 13q14 in 29 patients (37%) between D13S153 and the AFMa 301wb5. Classical cytogenetics was less sensitive, as it detected the 13q14 deletion in only one out of 69 patients (1%) in whom adequate metaphases were obtained. We found no significant difference in survival between patients with and patients without 13q14 LOH. In subset of patients with low beta2-microglobulin levels, those with 13q14 LOH had significantly shorter survival than did patients with low beta2-microglobulin levels but no 13q14 LOH. Also patients in early Rai stages (0-II) with 13q14 LOH had shorter survival period (P = 0.05) than did patients without LOH. These data confirm the prevalence of 13q14 deletion in CLL and suggest that this deletion may help identify more aggressive disease in patients presenting with early stage disease. PMID- 10475619 TI - A colorimetric assay to evaluate the chemotherapeutic response of children with acute lymphoblastic leukemia (ALL) employing achatininH: a 9-O-acetyl sialic acid binding lectin. AB - Employing a 9-O-acetyl sialic acid binding lectin, Achatinin(H) (ATNH), we have reported a non-invasive, blood based lymphoproliferation assay which measures the maximal lymphoproliferative dose (MLD) of ATN(H) to assess the status of 9-O acetylated sialoglycoconjugates (9-OAcSGs) in patients with Acute lymphoblastic leukemia (ALL) (Mandal C, Sinha D, Sharma V, Bhattacharya DK. O-acetyl sialic acid binding lectin, as a probe for detection of subtle changes on the cell surface induced during acute lymphoblastic leukemia [ALL] and its clinical application. Ind J Biochem Biophys 1997;34:82; Sinha D, Mandal C, Bhattacharya DK. Development of a simple blood based lymphoproliferation assay to assess the clinical status of patients with acute lymphoblastic leukemia. Leuk Res 1999;13:309-312; Sinha D, Mandal C, Bhattacharya DK. A novel method for prognostic evaluation of childhood acute lymphoblastic leukemia. Leukemia 1999;13[in press]). Although the expression of 9-OAcSGs clearly serves as an index of treatment outcome, the assay has limitations in that it requires radioisotopes, i.e. [3H]-TdR. Therefore a colorimetric assay was developed as an alternative approach. The pre-treatment MLD, as measured by the colorimetric assay, was 0.15 +/- 0.02 microg which progressively increased during consolidation therapy (1.40 +/- 0.39 microg), maintenance therapy (4.20 +/- 1.60 microg) and in followed-up cases (5.20 +/- 0.43 microg) but sharply declined following relapse (0.25 +/- 0.02 microg). The colorimetric assay also showed a good correlation with radiometric assay (r = + 0.93) and their mean coefficient of inter-assay precision were also comparable (15.53% versus 14.86%). We therefore propose that the colorimetric assay is a safe, non-radiometric, user friendly alternative for assessing individual chemotherapeutic responses in childhood ALL. PMID- 10475620 TI - Treatment of unfavorable myelodysplastic syndrome with all-trans-retinoic acid and subcutaneous Ara C. AB - Thirty patients with high-risk myelodysplastic syndrome, defined as > 5% bone marrow blasts, were treated with a combination of oral all-trans-retinoic acid (45 mg/m2 daily) and subcutaneous AraC (10 mg/m2) on days 1-14 of each 28-35 day cycle repeated for 2-6 cycles. Complete remission lasting 9, 12, and 15 months was achieved in three patients. Partial and minor response did not translate into meaningful clinical improvement, like complete responders. Overall incidence of leukemia transformation and survival of this cohort of patients was no different from the expected outcome for a group of patients with similar characteristics. PMID- 10475621 TI - Delphi-panel analysis of appropriateness of high-dose therapy and bone marrow transplants in chronic myelogenous leukemia in chronic phase. AB - BACKGROUND: It is uncertain which people with chronic myelogenous leukemia (CML) in chronic phase should receive conventional treatment (interferon and/or chemotherapy) versus high-dose therapy and a bone marrow transplant. There are no randomized trials comparing these approaches and analyses of data from non randomized studies are complex, contradictory without sufficient detail to allow subject-level treatment decisions. OBJECTIVE: Determine appropriateness of high dose therapy and bone marrow transplants in persons with CML in chronic phase with specific features. Develop a treatment algorithm. PANELISTS: nine leukemia experts from diverse geographic sites and practice settings. EVIDENCE: Boolean MEDLINE searches of chronic myelogenous leukemia and chemotherapy and/or transplants. CONSENSUS PROCESS: We used a modified Delphi-panel group judgment process. Age, prognostic score, disease duration, and type of conventional therapy and response were permuted to define 90 clinical settings. Each panelist rated appropriateness of high-dose therapy and a transplant versus conventional therapy on a 9-point ordinal scale (1, most inappropriate, 9, most appropriate) considering three types of donors: (1) HLA-identical siblings; (2) alternative donors (HLA-matched related or unrelated people other than an HLA-identical sibling); and (3) autotransplants. An appropriateness index was developed based on median rating and amount of disagreement. Relationship of appropriateness indices to permuted clinical variables was considered by analysis of variance and recursive partitioning. Preference between donor types was analyzed by comparing mean appropriateness indices of similar settings and a treatment algorithm developed. CONCLUSIONS: In people with CML in chronic phase and an HLA-identical sibling donor and in those with an alternative donor (but no HLA-identical sibling), a transplant was rated appropriate in those with a < or = partial cytogenetic response to interferon and uncertain or inappropriate in all other settings. Autotransplants were rated uncertain or inappropriate in all settings. Most of the variance in appropriateness ratings between different clinical settings was accounted for by response to interferon: complete versus < or = partial response. An HLA-identical sibling donor, when available, was always preferred to an alternative donor or autotransplant. In people without an HLA identical sibling, an alternative donor was favored over an autotransplant at higher appropriateness indices and the converse at lower appropriateness indices. PMID- 10475622 TI - Detection of apoptosis in acute promyelocytic leukemia cells in vivo during differentiation-induction with all-trans retinoic acid in combination with chemotherapy. AB - In two patients with acute promyelocytic leukemia (APL) treated with all-trans retinoic acid (ATRA) in combination with chemotherapy, we demonstrated that cells with apoptotic morphology were recognized in a small fraction of whole blood cells (0.2-0.4%) at the regression phase of leukocytosis with mostly maturing aberrant granulocytes of APL clone origin. Moreover, in a light-density cell fraction ( < 1.077) of peripheral blood or marrow cells obtained from three patients, DNA fragmentation was detected by agarose gel electrophoresis temporarily. These findings may suggest that, during the treatment with ATRA, a small fraction of APL cells maturing toward the stage of terminal differentiation underwent apoptosis in vivo, which might be enriched by the combination of chemotherapy to ATRA administration. PMID- 10475623 TI - A novel B-CLL specific nuclear protein (p44/46). AB - Our previous results indicated some diversities in electrophoretic patterns of proteins from different cellular fractions, i.e. nuclear, mitochondrial, microsomal and cytosolic isolated from mononuclear cells from the peripheral blood of B cell chronic lymphocytic leukemia (B-CLL) patients and healthy donors. Major differences were observed in electrophoretic banding of nuclear proteins from normal and transformed cells, especially in molecular mass region of 37 52 kDa. Electrophoretically-specific nuclear protein with molecular mass of 44/46 kDa of cells originating from B-CLL patients was used for raising polyclonal antiserum. As it was determined by Western blot technique (with alkaline phosphatase) obtained antiserum recognized 44/46 kDa antigen of nuclear fraction from B-CLL and acute lymphoblastic leukemia (ALL) cells, but not from normal ones. Our preliminary data were revealed that this antiserum shows no crossreactivity with leukemic nuclear proteins of patients with T cell chronic lymphocytic leukemia (T-CLL) and neither with nuclear polypeptides from either normal or cancerous (adenocarcinoma) stomach and colon mucosa. Immunological analysis was shown that higher expression of this particular antigen seems to correlate with progression of B-CLL. PMID- 10475624 TI - Constitutive protein expression of monocyte chemotactic protein-1 (MCP-1) by myelomonocytic cell lines and regulation of the secretion by anti- and proinflammatory stimuli. AB - We have investigated the protein expression of the chemokine monocyte chemotactic/chemoattractant protein-1 (MCP-1) in various human myelomonocytic leukemia cell lines. Applying specific ELISA, we demonstrated that this chemokine is produced constitutively by the cell lines HL-60, ML-2, MONO-MAC-6 and MUTZ-3 ranging between 440 and 1400 pg/ml MCP-1 per million cells. In the culture medium of two other unstimulated cell lines, MONO-MAC-1 and THP-1, almost no MCP-1 was detected. Stimulation of HL-60 and MONO-MAC-6 with lipopolysaccharide (LPS), and stimulation of ML-2 and MUTZ-3 with 12-tetradecanoyl phorbol 13-acetate (TPA) dramatically increased the MCP-1 level in the culture medium. The highest amount of MCP-1 (> 80 ng/ml within 24 h) was achieved by TPA stimulation of MUTZ-3 cells. Out of 15 cytokines tested for induction or enhancement of MCP-1 secretion, interleukin-3 (IL-3), IL-6, interferon-gamma (IFN-gamma), granulocyte macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF) and tumor necrosis factor (TNFalpha) were able to augment (twofold to 12-fold) the MCP-1 level in the culture medium of MONO-MAC-6 cells. While the antinflammatory cytokines IL-4, IL-10 and IL-13 failed to suppress MCP-1 secretion, the glucocorticoid dexamethasone strongly inhibited the MCP-1 production of unstimulated and stimulated MONO-MAC-6 cells. Thus, several regulatory elements are involved in MCP-1 secretion. Despite the quantitative differences of MCP-1 production among the cell lines analyzed, our results demonstrated a constitutive secretion in differentiation-arrested myelomonocytic leukemia cell lines and emphasize the usefulness of these malignant cell lines as models to study MCP-1 secretion and regulation. PMID- 10475626 TI - Apoptosis and its role in myelodyslastic syndrome. PMID- 10475627 TI - Behavioural responses of cockerels to semen collection and their influence on semen characteristics. AB - One-hundred-and-twenty cockerels of Strain L2 Taiwan Country chicken from the National Chung-Hsing University were tested for their behavioural responses to semen collection at 21 to 30, 34 to 43 and 57 to 66 weeks of age. At every age period, semen was collected from every cockerel 4 times within a 1-week period. 2. Cockerel performances on most semen traits, including volume, concentration, total number of sperm, ease of collection and sensitivity to massage were very consistent within each age period. 3. Males that did not respond nervously to handling or were less sensitive to massage appeared to produce less semen. However, contamination was more serious in males that responded very nervously or were very sensitive to massage. 4. Sensitivity to massage, like semen volume, is an invariant character as indicated by the high correlation between measures taken at different age periods. PMID- 10475625 TI - Chronic myeloid leukemia with expression of ALL-type BCR/ABL transcript: a case report and review of the literature. AB - CML with exclusive expression of ALL-type bcr/abl has only been rarely described. In some cases, the presence of this fusion gene has been associated to a differentiated subtype of CML that share some features with CMML, while in another case this molecular hallmark has been associated to a bad prognosis of the disease with a blast phase as clinical presentation or an early transformation to blast phase. We report a case of a 30-year-old woman who was diagnosed of CML in chronic phase in May 1989. She received treatment first with busulfan, achieving hematological remission and afterwards with interferon and Hydroxiurea. In February 1998, she was admitted at our hospital for an ABSCT. Then, molecular studies were performed. Multiplex PCR revealed the presence of a 481 bp product identified as the ela2 bcr/abl transcript and confirmed by sequencing. After 9 years from diagnosis, the patient remains in hematological remission and in good clinical condition. PMID- 10475628 TI - Influence of stocking density on behavioural, health and productivity traits of turkeys in large flocks. AB - 1. The effect of stocking density on welfare traits of turkeys was studied in 2 experiments. In each experiment 2,633 sexed BUT turkey poults were assigned to 3 rooms, 135 m2 with 1 treatment per room. Because of the large flock size (675,878 and 1080 birds in T1, T2 and T3 respectively) treatments were not replicated 2. Floor space allowances varied according to treatment; for the males: 24 dm2, 18.5 dm2 and 15 dm2 until week 12 and 40 dm2, 31 dm2 and 25 dm2 from week 12; for the females: 16 dm2, 12.3 dm2 and 10 dm2. 2. The scan sampling method in experiment 1 and the focal sampling method in experiment 2 were used to record behaviour at week 6, 9, 12 (males and females), and 16 (males). Birds' ethogram was divided into 7 mutually exclusive behaviours: standing/walking, resting, feeding, drinking, pecking at the environment, pecking at another bird, and preening. Position changes in the pen and the frequency of disturbances of resting birds by other birds were recorded in experiment 2. 3. Gait was assessed at week 12 (females) and week 16 (males). Prevalence of lesions, breast (males), hip and foot (males and females), were recorded at slaughter. Birds were weighed at week 12 (males and females) and week 16 (males). 4. Stocking density had little influence on behaviour except on the frequency of disturbances of resting birds by other birds, which tended to be more frequent at the highest density. 5. Gait deteriorated as stocking density increased. Hip and foot lesions were more frequent at the highest density. Bodyweight decreased significantly with decreasing floor space. 6. The results suggest that turkey welfare was poorer at the highest density than at the 2 lower stocking densities. PMID- 10475629 TI - Spectral sensitivity of the domestic fowl (Gallus g. domesticus). AB - 1. The spectral sensitivity of 7 broiler fowl was determined in a behavioural test. 2. Initially the birds were trained to receive a food reward by pecking at a clear, Perspex panel behind which was a light stimulus (circular, diameter = 9 mm), originating from a tungsten-halogen lamp. Subsequently, they were trained to choose between 2 panels only 1 of which was lit; the assignation of light and dark on each panel was randomly ascribed between trials. The colour of the lit panel was determined by the wavelength of the light transmitted through one of 13 closely defined narrow bandwidth filters between 32665 years) were randomized to receive once daily medication for 4 days with a 16-day interval between treatment periods. Assessments of attention, memory and motor control were made prior to dosing on each day, at 2, 4, 6 and 8 h after dosing on days 1 and 4, and at 24 and 48 h following the last dose. On day 1, detectable impairment was observed at all time points in both groups. On day 4, haloperidol treated subjects showed increased impairment compared with day 1 and this was sustained throughout the 48 h of testing. Olanzapine treated subjects showed reduced day 4 deficit (compared with day 1), with no significant difference from placebo beyond 6 h post dose. These results suggest that both haloperidol and olanzapine have a measurable initial effect on cognitive and psychomotor function in elderly volunteers. However, acute effects associated with olanzapine decrease with repeated dosing and show substantial adaptation within 4 days. In contrast, effects seen with haloperidol are sustained and increase with repeated dosing over the same period. PMID- 10475722 TI - Single dose pharmacodynamics of thioridazine and remoxipride in healthy younger and older volunteers. AB - Phenothiazines are widely used in older patients, but little experimental work has been carried out in this age group. Two groups of healthy volunteers, a younger group (Y: six males and six females, aged 20-42 years) and an older group (O: six males and eight females, aged 65-77 years) took part in a randomized double-blind three-period crossover study in which they received by mouth single doses of thioridazine (Y: 50 mg; O: 25 mg) remoxipride (Y: 100 mg; O: 50 mg) or placebo. Measures of central nervous system (CNS) and haemodynamic function were carried out before drug administration and at 1.5-h intervals up to 9 h post dose, and blood samples were collected over a 24-h period. No significant differences in dose-corrected pharmacokinetic variables were found between the two groups. There was evidence of marked CNS depressant effects of thioridazine from both objective and subjective measures. The effects for remoxipride were similar, though generally less marked. After allowance was made for dose, there was little indication of any difference in degree of CNS depression between the two age groups. Haemodynamic measures showed orthostatic reductions in blood pressure with thioridazine which were particularly marked in the older group, who also showed lower compensatory increases in pulse rate. These results indicate potential problems with orthostatic hypotension with thioridazine in older patients. CNS depression may also be a problem, especially in patients with compromised cholinergic function. PMID- 10475723 TI - The 5-HT1A receptor agonist flesinoxan increases aversion in a model of panic like anxiety in rats. AB - Acute systemic administration of the selective serotonin (5-HT)1A receptor full agonist flesinoxan enhanced the sensitivity of rats to the panic-like aversion elicited by local stimulation of the dorsolateral periaqueductal grey (dPAG). This experimental paradigm in rats has previously been validated as a simulation of acute anxiety with particular relevance to panic disorder. The dose-dependent decrease in threshold for acute fear responses recorded in rats following intraperitoneal administration of flesinoxan (1-10 mg/kg) was similar to that induced by the panic precipitating agent yohimbine and opposite to the threshold increase induced by the antipanic drug alprazolam. The proaversive effect of flesinoxan observed in rats is consistent with the reported aggravation of the condition of panic patients following oral flesinoxan treatment. Thus, the model adequately detects drug-induced panicogenic-like properties. Data suggest that selective activation of 5-HT1A receptors (pre- and/or post-synaptic in brain and/or periphery) following systemic administration of 5-HT1A receptor full agonists exacerbates aversion in animals or patients with panic anxiety; activation of these receptor subtypes may probably mediate the panicogenic action reported under certain circumstances with non-selective 5-HT mimetics. PMID- 10475724 TI - Effects of D-cycloserine, a positive modulator of N-methyl-D-aspartate receptors, and ST 587, a putative alpha-1 adrenergic agonist, individually and in combination, on the non-delayed and delayed foraging behaviour of rats assessed in the radial arm maze. AB - The present study investigated whether alpha-1 adrenergic and glutamatergic N methyl-D-aspartate (NMDA) receptor-mediated mechanisms interact in memory processes, by examining the effects of individual and combined systemic administration of ST 587, a putative alpha-1 agonist, and D-cycloserine (DCS), a partial agonist at the glycine-B binding site of the NMDA receptor, on the performance of rats in non-delayed and delayed (4-6 h) foraging behaviour in the radial arm maze task, using the delayed non-matching to sample (DNMTS) version. The results indicated that DCS (5.0 mg/kg) decreased working memory errors, i.e. the number of re-entries into the previously visited arms during the sampling phase. In addition, both ST 587 (100 microg/kg) and DCS (10 mg/kg), when administered alone 30 min before a sampling phase, improved retention of this task as reflected by the increased number of correct choices before the first error during the retention phase. The combined administration of ST 587 and DCS, however, did not lead to better retention in the DNMTS task compared with the administration of each of the drugs alone. Combinations of sub-threshold doses of ST 587 (50 or 75 microg/kg) and DCS (5.0 or 7.5 mg/kg) also did not improve retention in this task. DCS (5.0 or 7.5 mg/kg) increased activity as indicated by the increased number of arm entries in a given time during the sampling phase. These findings suggest that the systemic administration of a positive modulator of the NMDA receptor facilitates hippocampal-dependent memory functions, but that these effects are not enhanced by combined administration with an alpha-1 agonist, even though the alpha-1 agonist is effective when given alone. The results support the idea that NMDA receptors modulate both mnemonic and non mnemonic functions in the brain. PMID- 10475725 TI - Impulsivity: a discussion of clinical and experimental findings. AB - Impulsivity can often be an important clinical problem in psychiatry and neurology. In psychiatry, the manifestation of impulsive behaviour in syndromes such as personality disorders, attention deficit hyperactivity disorder and in substance abuse may be different, and this has led to conflicting definitions. There has also been a tendency to concentrate on the nature of the behavioural manifestation (problems with the law, aggression, drug use, behavioural problems in school) rather than shared psychological processes, and to ignore the fact that impulsivity can also have positive aspects. In a normal population, the personality trait of impulsivity has been analysed using personality inventory questionnaires. Analysis of these data lead to the suggestion that impulsivity as commonly defined and understood may be made up of several independent factors, which may have separate biological bases. These self-rating questionnaires have been complemented by objective tests that are now often computerized, and which have been used in man (e.g. with criminal offenders, children, or patients who have undergone brain surgery). Some of these tests, such as the differential reinforcement of low rates procedure or the delay of reinforcement procedure, have also been used to study impulsivity in animals. Analysis of the behavioural principles of these tests suggests that they too may reflect different aspects of impulsivity. Many different biological systems have been proposed to contribute to the neurobiological basis of impulsivity. The serotonergic neurotransmitter system has recently received the most attention, with evidence of its involvement coming from animal studies as well as from studies in psychiatric patients. The frontal lobes have been proposed to play an important role in regulating impulsivity, although it unclear how specific this is. None of this biological knowledge has yet led to reliable pharmacotherapy for excessive impulsivity and, as yet, there is little understanding of the mechanisms by which those drugs, which have been found empirically to have some efficacy (e.g. the psychomotor stimulants in attention deficit hyperactivity disorder), exert their therapeutic effect. By bringing together knowledge from different areas of research it is hoped that a cross fertilization will be achieved, which will lead to a sharpening of concepts, an improvement in methodology and the stimulation of biological studies. PMID- 10475726 TI - An acute ischaemic event associated with the use of venlafaxine: a case report and proposed pathophysiological mechanisms. AB - Venlafaxine, a structurally novel antidepressant, belongs to a new generation of antidepressants--the serotonin/noradrenaline reuptake inhibitors. In clinical trials, venlafaxine was found safe and effective in most patients. However, significant changes in vital signs (hyper- and hypotension) and cardiac conduction abnormalities were observed in a few patients, notably in elderly patients. We present a case of an elderly woman with a pre-existing history of ischaemic heart disease, who was treated with venlafaxine, and developed acute myocardial ischaemia within the first week of treatment. This is the first report of a possible association between an acute cardiovascular event and venlafaxine. The association of venlafaxine treatment with ischaemic events could be explained by its unique pharmacological and haemodynamic properties. PMID- 10475727 TI - The use of dexamethasone in elderly patients with antidepressant-resistant depressive illness. AB - Many depressed patients do not respond to first-line antidepressant treatment. Dexamethasone is a synthetic steroid which may have antidepressant properties. Its use in two elderly patients with resistant depression is reported. Both patients appeared to benefit from the treatment. The possible modes of action of this treatment, and its potential benefits to the elderly, are discussed. PMID- 10475728 TI - Olanzapine for mixed episodes of bipolar disorder. PMID- 10475729 TI - Effects of bromocriptine and haloperidol on prepulse inhibition of the acoustic startle response in man. PMID- 10475730 TI - Rapid tranquillisation in isolated units, i.m. medication preferable to i.v. PMID- 10475731 TI - Dopamine receptors, antipsychotic action and schizophrenia. PMID- 10475732 TI - Papers resulting from presentations made at the 1998 European Society for Clinical Virology meeting: Progress in Clinical Virology IV, held in Hamburg, Germany. PMID- 10475733 TI - [Informed consent in nuclear medicine]. PMID- 10475734 TI - [Radioisotopic studies in the diagnosis of obstructive uropathy]. PMID- 10475735 TI - [Radiolabelling of human IgG with ultraviolet light having its own reactor]. PMID- 10475737 TI - Modification of blood-pressure-measuring devices and the protocol of the British Hypertension Society. PMID- 10475736 TI - [Thyroid scintigraphy]. PMID- 10475738 TI - Potency and bioavailability of therapeutic formulations during their field usages. PMID- 10475739 TI - USAN Council. List No.418. New names. Edodekin alfa. PMID- 10475740 TI - USAN Council. List No.418. New names. Etanercept. PMID- 10475741 TI - Proceedings of the International Symposium on The Atopy Syndrome in the 3rd Millennium. Zurich, Switzerland, October 2-3, 1998. PMID- 10475742 TI - Recent advances in the healthy effects of nonalcoholic wine compounds. Proceedings of a meeting. Florence, Italy, November 26-28, 1998. PMID- 10475743 TI - Is it better to find Adam or study spermatogenesis? --a difference opinion. PMID- 10475744 TI - Your genome--just another credit card to carry around. PMID- 10475745 TI - An unusual pattern of invertase activity development in the thermophilic fungus Thermomyces lanuginosus. AB - In the thermophilic fungus Thermomyces lanuginosus, invertase displays an unusual pattern of development: the induced activity begins to diminish even before any substantial quantity of sucrose has been utilized or an appreciable amount of biomass has been produced. Despite this pattern of invertase activity, neither the growth rate nor the final mycelial yield is affected adversely. T. lanuginosus invertase is a thiol protein and the enzyme is active when specific sulfhydryl group(s) is in the reduced state. Measurements of reduced coenzyme and glutathione pools in sucrose-growth mycelia excluded oxidative stress as the primary reason for the observed decline in invertase activity. Rather, this unusual pattern of invertase is considered to be due to its localization in the hyphal tips. At the early stage of growth, the number of hyphal tips per unit mass of mycelium is maximum, whereas at later times their numbers do not increase in proportion to the biomass. As a result invertase activity shows an apparent inverse relationship with biomass. The enzyme activity disappears when the inducing carbon source is consumed and growth is completed. PMID- 10475746 TI - Cellular mechanisms and molecules in infection immunity. Papers from the 5th Elsinore Meeting on Infection Immunity. Elsinore, Denmark, 29 May-2 June 1998. PMID- 10475747 TI - Proceedings of the 2nd International Conference Novel Approaches to the Control of the Helminth Parasites of Livestock. Baton Rouge, Louisiana, USA. 22-26 March 1998. PMID- 10475748 TI - [Progress and continuing education in medicine. 1: Cancer diagnosis, therapy, and after care]. PMID- 10475749 TI - Stability of IS6110 restriction fragment length polymorphism patterns of multidrug-resistant Mycobacterium tuberculosis strains. PMID- 10475750 TI - Monoclonal antibody 13F1 produces annular immunofluorescence patterns on Cryptococcus neoformans serotype AD isolates. PMID- 10475751 TI - A plea for caution with regard to applicability of PCR for direct detection. PMID- 10475752 TI - Intestinal myiasis caused by Eristalis tenax. PMID- 10475753 TI - Dialogue or monologue? The limits to lessons that can be learned. PMID- 10475754 TI - Pseudotumour after arteriovenous malformation embolisation. PMID- 10475755 TI - False negative polymerase chain reaction on cerebrospinal fluid samples in tuberculous meningitis established by culture. PMID- 10475756 TI - False negative polymerase chain reaction on cerebrospinal fluid samples in tuberculous meningitis. PMID- 10475757 TI - A novel mutation of the myelin P(o) gene segregating Charcot-Marie-Toothdisease type 1B manifesting as trigeminal nerve thickening. PMID- 10475758 TI - Intracranial extracerebral follicular lymphoma mimicking a sphenoid wing meningioma. PMID- 10475759 TI - Determinants of the copper concentration in cerebrospinal fluid. PMID- 10475760 TI - Solitary intracranial myofibroma in a child. PMID- 10475761 TI - Axonal polyneuropathy and encephalopathy in a patient with verotoxin producing Escherichia coli (VTEC) infection. PMID- 10475762 TI - Crying spells as symptoms of a transient ischaemic attack. PMID- 10475763 TI - Continuous drop type of orthostatic hypotension. PMID- 10475764 TI - Respiratory aspects of neurological disease. PMID- 10475765 TI - Idiopathic cerebellar ataxia associated with celiac disease: lack of distinctive neurological features. PMID- 10475766 TI - Procainamide for faecal incontinence in myotonic dystrophy. PMID- 10475767 TI - Flail arm syndrome of Vulpian-Bernhart's form of amyotrophic lateral sclerosis. PMID- 10475768 TI - Pain after whiplash. PMID- 10475769 TI - Adenosine and cardioprotection in the diseased heart. AB - Biological and mechanical stressors such as ischemia, hypoxia, cellular ATP depletion, Ca2+ overload, free radicals, pressure and volume overload, catecholamines, cytokines, and renin-angiotensin may independently cause reversible and/or irreversible cardiac dysfunction. As a defense against these forms of stress, several endogenous self-protective mechanisms are exerted to avoid cellular injury. Adenosine, a degradative substance of ATP, may act as an endogenous cardioprotective substance in pathophysiological conditions of the heart, such as myocardial ischemia and chronic heart failure. For example, when brief periods of myocardial ischemia precede sustained ischemia, infarct size is markedly limited, a phenomenon known as ischemic preconditioning. We found that ischemic preconditioning activates the enzyme responsible for adenosine release, ie, ecto-5'-nucleotidase. Furthermore, the inhibitor of ecto-5'-nucleotidase reduced the infarct size-limiting effect of ischemic preconditioning, which establishes the cause-effect relationship between activation of ecto-5' nucleotidase and the infarct size-limiting effect. We also found that protein kinase C is responsible for the activation of ecto-5'-nucleotidase. Protein kinase C phosphorylated the serine and threonine residues of ecto-5' nucleotidase. Therefore, we suggest that adenosine produced via ecto-5' nucleotidase gives cardioprotection against ischemia and reperfusion injury. Also, we found that plasma adenosine levels are increased in patients with chronic heart failure. Ecto-5'-nucleotidase activity increased in the blood and the myocardium in patients with chronic heart failure, which may explain the increases in adenosine levels in the plasma and the myocardium. In addition, we found that further elevation of plasma adenosine levels due to either dipyridamole or dilazep reduces the severity of chronic heart failure. Thus, we suggest that endogenous adenosine is also beneficial in chronic heart failure. We propose potential mechanisms for cardioprotection attributable to adenosine in pathophysiological states in heart diseases. The establishment of adenosine therapy may be useful for the treatment of either ischemic heart diseases or chronic heart failure. PMID- 10475770 TI - Quantitative analysis of surface P-wave morphology in isthmus ablation for type 1 atrial flutter: differentiation between complete isthmus block and slow isthmus conduction. AB - Changes in P-wave morphology in inferior leads during atrial pacing at the margins of the carvo-tricuspid isthmus have been reported to be useful for predicting the creation of isthmus block in radiofrequency (RF) ablation of type I atrial flutter (AFL). However, it is not known whether these changes in P-wave morphology allow the clinician to differentiate between complete isthmus block and slow isthmus conduction. P-wave morphology during low lateral right atrial (LLRA) pacing, as well as during coronary sinus ostium (PCS) pacing, was evaluated prior to ablation, during slow isthmus conduction, and after complete isthmus block in 30 patients with AFL. Changes in P-wave morphology during LLRA pacing were not sufficient to differentiate between complete isthmus block and slow isthmus conduction. While changes in P-wave morphology in lead II from inverted to biphasic during PCS pacing were observed in both slow isthmus conduction and complete isthmus block, the ratio of the positive component to the total P-wave amplitude (P-wave ratio) was significantly different between slow isthmus conduction (20+/-17%) and complete isthmus block (40+/-11%) (P<0.0001). When the P-wave ratio in lead II during PCS pacing was more than 75% of the F wave ratio in lead II during AFL, bilateral complete isthmus block was predicted with a sensitivity of 88%, a specificity of 71%, a positive predictive value of 75%, and a negative predictive value of 85%. These results indicate that a P-wave ratio greater than 20% or a P-wave ratio during PCS pacing greater than 75% of the F-wave ratio during AFL may predict a bidirectional complete isthmus block. PMID- 10475772 TI - Increased exhaled nitric oxide and impaired oxygen uptake (VO2) kinetics during exercise in patients with chronic heart failure. AB - Vascular endothelial function is abnormal in patients with congestive heart failure (CHF). Exhaled nitric oxide (NO) output is a marker of pulmonary endothelial NO release. The present study examined the relation between exhaled NO output and oxygen uptake (VO2) kinetics at the onset of exercise, which reflects blood flow response. Sixteen patients with CHF and 7 volunteers underwent constant bicycle exercise. Oxygen deficit and time constant for VO2 increment at the onset of exercise were analyzed. Exhaled NO concentration was measured by a chemiluminescence analyzer and exhaled NO output was calculated by multiplexing ventilation. Exhaled NO output was significantly greater in the CHF group than in the control group at rest (86+/-65 nl min(-1) m(-2) vs 298+/-135 nl min(-1) m(-2), p<0.001) and during exercise (152+/-98 nl min(-1) m(-2) vs 455+/ 190 nl min(-1) m(-2), p<0.001). However, the %increase of NO output was significantly smaller in the CHF group than in the control group (70+/-26% vs 109+/-85%, p<0.05). Oxygen deficit was significantly greater in the CHF group than in the control group (240+/-70 ml vs 372+/-107 ml, p<0.01) and the time constant for VO2 increment was also significantly prolonged in the CHF group (35.1+/-8.0 s vs 50.1+/-16.3 s, p<0.05). Exhaled NO output during exercise significantly correlated with oxygen deficit (r=0.67, p<0.001) and the time constant for VO2 increment (r=0.74, p<0.001). Increased NO output played a counter-regulatory role in the impaired blood flow in CHF. PMID- 10475771 TI - Expression of cytokine and adhesion molecule mRNA in atherectomy specimens from patients with coronary artery disease. AB - Coronary arteriosclerosis is an underlying condition in acute myocardial infarction (AMI), unstable angina pectoris (UAP) and stable angina pectoris (SAP), and is also related to restenosis (RS) following coronary intervention. To investigate the pathogenesis of this condition, a quantitative reverse transcriptase polymerase chain reaction was used to determine relative levels of mRNA for interleukin (IL)-1beta, IL-6, IL-8, transforming growth factor beta (TGF beta), intercellular adhesion molecule (ICAM)-1, E-selectin and vascular cell adhesion molecule (VCAM)-1 using directional coronary atherectomy (DCA) specimens. Eleven patients with AMI, 7 with UAP, 10 with SAP and 6 with RS following a previous coronary intervention underwent DCA. The mRNA intensity for each molecule was expressed by comparing it with that of beta-actin mRNA. The AMI and UAP patients showed high frequencies of mRNA for IL-1beta, IL-8, TGF-beta, and ICAM-1 together with strong intensities of expression, whereas SAP patients showed decreased mRNA expression for these molecules. Increased IL-6 mRNA expression was observed only in AMI samples. Specimens from RS patients revealed an accumulated expression of proinflammatory cytokines, except for IL-6, as well as of TGF-beta. The study suggests that variation in mRNA expression may reflect the pathophysiology of specific types of coronary artery disease, and remodeling following vascular injury. PMID- 10475773 TI - Mechanism of increase in anaerobic threshold during recovery phase in patients with acute myocardial infarction. AB - The contribution of venous return to the increase in anaerobic threshold (AT) in patients with acute myocardial infarction (AMI) was investigated. Twenty-seven patients with uncomplicated AMI underwent supine and sitting cardiopulmonary exercise testing, and 10 out of these 27 patients performed constant-workload tests in which cardiac output before and after a phase II program was measured. Data from 8 patients were eliminated because of restenosis after direct percutaneous transluminal coronary angioplasty. The increase in AT was significantly greater in the sitting position than in the supine position, 15.0+/ 9.9% and 6.3+/-6.0% respectively. Average O2 pulse at AT changed non significantly from 8.6+/-1.5 to 8.7+/-1.2 ml min(-1) beat(-1) in the supine position, but it showed a significant increase from 8.1+/-1.3 to 9.2+/-1.3 ml min(-1) beat(-1) when measured in the sitting position. In the constant-workload studies, stroke index showed a significant increase during both supine and sitting exercise. Percent increase in the stroke index from rest to exercise did not differ significantly in the supine position, but did differ significantly in the sitting position. These results strongly suggest that increased venous return as well as improvement in cardiac pump function play a major role in the mechanism of AT increase in an upright position throughout the recovery phase in AMI patients. PMID- 10475775 TI - The time from anaerobic threshold (AT) to respiratory compensation point reflects the rate of aerobic and anaerobic metabolism after the AT in chronic heart failure patients. AB - The significance of the time from anaerobic threshold to respiratory compensation point (RCP-AT time) in patients with chronic heart failure was investigated. Thirty-seven patients with chronic heart failure (New York Heart Association class II or III) were enrolled into the study. Cardiopulmonary exercise testing was performed using breath-by-breath gas sampling. A bicycle ergometer was used, and incremental exercise testing was carried out. Anaerobic threshold, respiratory compensation point (RCP), and the slope of oxygen uptake (VO2) as a function of work rate (deltaVO2/deltaWR) were measured. A positive correlation (r=0.53) between RCP-AT time and deltaVO2/deltaWR was found. RCP-AT time was corrected for the whole exercise period (ramp exercise-RCP point), and the correlation between corrected RCP-AT time and deltaVO2/deltaWR was still present (r=0.46). There was no correlation between RCP-AT time and anaerobic threshold. These findings suggest that RCP-AT time is a new parameter that reflects the rate of the aerobic and anaerobic metabolism after AT. PMID- 10475774 TI - Magnesium dynamics and sympathetic nervous system activity in patients with chronic heart failure. AB - This study was undertaken in patients with heart failure to investigate the relation between plasma norepinephrine (NE) concentration and Mg dynamics. The study subjects comprised 16 patients with chronic heart failure (mean age 64.9+/ 10.0 years). Cardiopulmonary exercise testing was performed on all patients, and anaerobic threshold (AT), peak oxygen uptake (peak VO2) and peak exercise time were measured. Resting and peak values of plasma NE concentration and serum and erythrocyte magnesium concentration were also measured. The results were as follows: the serum Mg concentration was increased significantly immediately after exercise (p<0.01), and the erythrocyte Mg concentration showed a tendency to decrease (p<0.1). The resting plasma NE level was inversely correlated with AT (p<0.05, r=-0.57), peak VO2 (p<0.05, r=-0.55) and peak exercise time (p<0.01, r= 0.62). When the plasma NE concentration at rest was analyzed in 2 groups of patients, ie, those with higher than average and those with lower than average concentrations, the resting erythrocyte Mg concentration was significantly lower in the high-NE group (2.2+/-0.3 mg/dl) than in the low-NE group (2.7+/-0.5 mg/dl) (p<0.05). The data indicate that patients with chronic heart failure associated with high NE levels at rest who showed low exercise tolerance have intracellular hypomagnesemia, which may be caused by Mg migration from intracellular to extracellular spaces. PMID- 10475776 TI - Time course of expression and localization of heat shock protein 72 in the ischemic and reperfused rat heart. AB - While the role of heat shock protein 72 (HSP72) in protection of the ischemic heart has been studied extensively, the expression and localization of HSP72 in the ischemic and reperfused heart remains poorly understood. This study examined the expression and distribution of HSP72 after various periods of ischemia and reperfusion in the non-stress-pretreated rat heart. The distribution of HSP72 in the myocardium was examined immunohistochemically using a specific monoclonal antibody for detection of this protein. The levels of HSP72 mRNA in the myocardium were estimated by the reverse transcriptase polymerase chain reaction (RT-PCR) method. Rats were subjected to either permanent occlusion of the left coronary artery or 30 min of occlusion followed by reperfusion, and they were sacrificed 30 min, 4 h, 24 h and 7 days thereafter. Both permanent ischemia and reperfusion induced expression of HSP72 mRNA in the ischemic myocardium from 30 min to 24 h after the onset of ischemia/reperfusion. The maximum mRNA level, which appeared at 4 h, was much higher in the reperfusion group than in the permanent ischemia group. Myocytes exhibited definite immunoreactivity of HSP72 at 4 h and 24 h in both groups, and microvessels exhibited strong immunoreactivity at 4 h in the reperfusion group. This study revealed the time course of expression and distribution of HSP72 in the ischemic and reperfused hearts in vivo. PMID- 10475777 TI - Photodynamic therapy of atherosclerosis using YAG-OPO laser and Porfimer sodium, and comparison with using argon-dye laser. AB - We performed photodynamic therapy (PDT) using the Yttrium Aluminium Garnet Optical Parametric Oscillated (YAG-OPO) laser in cases of atherosclerosis, and examined its efficacy in vivo. We also performed PDT using an Argon-dye (Ar-dye) laser with the same output, and compared the efficacies. Following balloon denudation injury of the thoracoabdominal aorta, rabbits were raised on a cholesterol diet for 16 weeks, producing atheroma in that region. At 24 h following the administration of Photofrin 5 mg/kg, PDT was performed, and animals were sacrificed at 1 day, 1 week, and 2 weeks following the procedure to examine its efficacy. This was compared with the efficacy of PDT using the Ar-dye laser. Following PDT using a YAG-OPO laser, an increase in the vessel lumen was seen due to reduction of the hypertrophic intima and media, without the appearance of inflammatory cells. This result was seen more strongly in PDT using the pulse wave YAG-OPO laser than with the continuous wave Ar-dye laser, affecting not just the intima but also the media. These data demonstrated that PDT can effectively regress atherosclerotic lesions. PMID- 10475778 TI - Gadolinium suppresses stretch-induced increases in the differences in epicardial and endocardial monophasic action potential durations and ventricular arrhythmias in dogs. AB - We tested whether acute pressure overloading of the left ventricle (LV) had spatially different effects on repolarization, thereby causing arrhythmias. The effects of gadolinium (Gd3+), a nonspecific blocker of stretch-activated channels were also examined. In anesthetized dogs, 5 s clamping of the ascending aorta (AC), separated by 5-min intervals, was repeated while monophasic action potentials (MAPs) were recorded from the LV endocardium and epicardium. Gd3+ was injected into the left atrium before the second (500 micromol) and third AC (2500 micromol) (n=10). In a separate group (n=7), the effects of Gd3+ in the presence of verapamil were examined. Epicardial MAP durations at 50% and 90% repolarization (APD50; APD90) shortened in response to LV pressure rise and elongation of the segment length induced by the first AC, whereas endocardial MAP durations remained unchanged. Thus, the difference in APD50 and APD90 increased. Consistent with these changes, premature ventricular contractions (PVCs) developed. Gd3+ had no effect on baseline MAP durations, however it prevented an AC-induced increase in the difference by suppressing epicardial MAP shortening. Gd3+ also reduced PVCs in a dose-dependent manner at plasma concentrations of 1-4 micromol/L. The effects were also evident after administration of verapamil. Thus, gadolinium suppressed an increase in the spatial dispersion of repolarization and arrhythmias via a mechanism of action different from that of verapamil. PMID- 10475779 TI - Effects of immunosuppressants on platelet-derived growth factor-A chain mRNA expression and coronary arteriosclerosis in rat cardiac allografts. AB - Graft coronary arteriosclerosis (GCA) that results in proliferative and obstructive lesions limits the long-term success of cardiac transplantation. Despite extensive study, the pathogenic mechanisms underlying GCA are still unclear and therapeutic strategies for this condition have been inadequate. In this study, we compared the therapeutic effectiveness of cyclosporine A (CsA), 15 deoxyspergualin (DSG), and Multiglycosidorum tripterygii (MT) on GCA. In addition, we studied the correlation between the extent of GCA and the degree of platelet-derived growth facter (PDGF)-A chain mRNA expression in cardiac grafts. Lewis rats receiving heterotropic heart transplants from Wistar King donors were treated with 10 mg kg(-1) day(-1) of CsA (n=7), 5 mg kg(-1) day(-1) of DSG (n=7) or 30 mg kg(-1) day(-1) of MT (n=7) respectively. Histological evaluation of coronary arteriosclerosis and Northern blot analysis of cardiac allograft PDGF-A chain mRNA expression were conducted on day 60 after transplantation. Varying levels of GCA were observed in the 21 transplanted hearts. Significant differences in both the degree of PDGF-A mRNA expression and the extent of GCA were found among the 3 groups. GCA was significantly reduced in allografts treated with MT or DSG in comparison with the level seen in CsA-treated grafts. A significant correlation was found between PDGF-A chain mRNA expression and the grade of arterial intimal thickening (r=0.76, p<0.05) as well as with the incidence of diseased vessels (r=0.82, p<0.01). Our results indicate that both MT and DSG are more effective in the treatment of GCA than CsA. In our cardiac allografts, the degree of PDGF-A chain mRNA expression correlated well with the extent of GCA, suggesting that PDGF-A may play an important role in the development of transplant-related GCA. PMID- 10475780 TI - Coronary artery bypass grafting in the acute phase after renal transplantation: report of a case. AB - To the best of our knowledge, only 3 cases of coronary artery bypass grafting (CABG) performed under cardiopulmonary bypass (CPB) on patients in the chronic phase after renal transplantation have been reported in Japan. The first case of a patient who underwent CABG in the acute phase after renal implantation in Japan is herein described. Perioperatively, oral immunosuppressive agents were discontinued and they were given intravenously. Cyclosporin A (Cy-A) was administered via a continuous intravenous infusion in the acute phase after renal transplantation and closely monitored, because the blood concentration of Cy-A can vary a great deal during the perioperative period. This case report serves to demonstrate that as long as appropriate immunosuppressive drugs are perioperatively administered, CABG under CPB can be safely performed on patients who have undergone renal transplantation without subsequent rejection, infection, or renal damage, even during the acute phase. PMID- 10475781 TI - A chronic, massive thrombus in the right main pulmonary artery: a case report and echocardiographic analysis. AB - A chronic, large thrombus in the right main pulmonary artery (PA) was detected in a 54-year-old woman with a history of surgical repair of atrial septal defect. Color flow imaging revealed a prominent red signal along the right border of the markedly dilated PA. Pulsed Doppler echocardiography showed that the red signal was caused by flow reversal occurring during systole. According to the physics of blood flow, flow reversal probably represents secondary, helical flow, which may be related to thrombus formation. PMID- 10475782 TI - A case in which stent insertion is considered to have triggered contrast medium induced coronary vasospasm. AB - A Gianturco-Roubin II (GR-II) stent was inserted in a 75-year-old man who developed restenosis of the right coronary artery (RCA) after percutaneous transluminal coronary angioplasty (PTCA). Although the vessel became partially occluded after 7 months, it was redilated by PTCA. Follow-up angiography of the RCA and left coronary artery (LCA) was performed 3 months later. Chest pain with bradycardia and hypotension occurred immediately after this examination, and ST elevation appeared in ECG leads II, III, and aVF. Repeat angiography of the RCA confirmed complete occlusion due to a spasm at a site proximal to the GR-II stent. The spasm was resolved by intracoronary infusion of isosorbide dinitrate (ISDN), and PTCA was carried out for extensive recurrent restenosis of the RCA; however, vascular dissection developed at the distal end of the GR-II stent. Therefore, a Palmaz-Schatz (P-S) stent was placed such that its proximal end overlapped the distal end of the GR-II stent. Follow-up angiography 3 months later showed no restenosis, but an episode of vasospasm similar to the previous one occurred immediately after left ventriculography. The RCA was completely occluded proximal to the GR-II stent because of spasm. Although this spasm was gradually relieved by intracoronary infusion of ISDN, marked spasm was also observed distal to the P-S stent; complete relief was achieved by infusion of additional ISDN. PMID- 10475783 TI - Scintigraphic evidence for a specific long-chain fatty acid transporting system deficit and the genetic background in a patient with hypertrophic cardiomyopathy. AB - The mechanism of cardiac uptake of long-chain free fatty acids has not been fully determined. We encountered a hypertrophic cardiomyopathy patient who showed a lack of cardiac uptake of 2 different types of long-chain fatty acid analogues on the scintigraphic images. Flow cytometric analysis revealed no platelet or monocyte CD36 molecule expression (type I CD36 deficiency) and his CD36 gene showed homozygous mutation for 478C to T substitution, leading to an abnormal CD36 amino acid sequence. These findings strongly suggest that a specific transporting system rather than a simple diffusion is commonly involved in the cardiac uptake of long-chain free fatty acids in humans, and that the CD36 protein is the most likely candidate for the specific transporter and to explain scintigraphic defects on fatty acid imaging. PMID- 10475784 TI - Hyperkalemia probably reverses the antiarrhythmic effects of amiodarone: a case report. AB - Sustained monomorphic ventricular tachycardia (VT) developed in a 58-year-old man with acute myocardial infarction and end-stage renal disease. Amiodarone was effective in preventing VT recurrence. Sustained VT was not induced during an electrophysiologic study. However, VT recurred during accidental hyperkalemia, which was caused by the change of dialysis therapy from peritoneal dialysis to hemodialysis. VT subsided with correction of hyperkalemia. Thereafter, VT did not recur as long as the serum potassium concentration was kept within the normal range. Several months later, the patient died suddenly because poor dietary compliance resulted in an increase in his potassium concentration. This case suggests that hyperkalemia may reverse the potent antiarrhythmic effects of amiodarone. PMID- 10475785 TI - Transcatheter embolization of arteriovenous malformations in Cowden disease. AB - A patient with Cowden disease and multiple arteriovenous malformations (AVMs) that resulted in high output heart failure is described. Cowden disease is a familial syndrome characterized by endodermal, mesodermal and ectodermal dysplasia causing benign and malignant tumors of the skin, breast, gastrointestinal tract, and thyroid gland. Our patient had gastrointestinal polyposis, a right renal tumor, a left lung tumor, an adenomatous goiter, and typical dermatologic findings such as facial papules, acral keratosis, gingival papillomatosis and hemangiomas. AVMs were observed in the pelvis, cervical vertebra, liver, and right supraclavicular area. Transcatheter embolization was performed 7 times for the pelvic AVMs, but the effect decreased with repetition and the patient died of heart failure 2 years after the first embolization. The serum levels of tissue plasminogen activator (t-PA), platelet-derived growth factor (PDGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), and transforming growth factor beta1 were high, suggesting that these angiogenic molecules may play a role in the pathogenesis of AVMs in Cowden disease. PMID- 10475786 TI - Primary pericardial synovial sarcoma with detection of the chimeric transcript SYT-SSX. AB - We report a case of a 19-year-old woman with a primary pericardial synovial sarcoma that extended from the right ventricular free wall to the posterior aspect of the left anterior thoracic wall. Synovial sarcoma was diagnosed by the detection of the chimeric transcript SYT-SSX using reverse transcriptase polymerase chain reaction (RT-PCR). This transcript is generated by reciprocal translocation between chromosomes X and 18, and is specific to synovial sarcoma that usually occurs in the extremities of young adults. When pathological and immunohistochemical diagnosis of synovial sarcoma is difficult, the molecular biological technique using RT-PCR becomes a powerful method of confirmation of this neoplasm. PMID- 10475787 TI - [Incidence of anesthesia-related complications in Japan during 1997]. PMID- 10475788 TI - [Present condition and problems of autopsy in japan]. PMID- 10475789 TI - Errors in reported association between Tamm-Horsfall protein and IgG. PMID- 10475790 TI - [Experimental study of artificial disk heart valves]. AB - The paper provides the results of an experimental study of domestic artificial EMIKS disk cardiac valves during simulated pulsing flow. The operation valves was examined in the aortic position by photochromic visualization. The patterns of flow velocities beyond the valves during the whole operating cycle were obtained. The models of EMIKS valves at opening angles of 65 degrees and 80 degrees and the model with acoustic clearance in the design of a disk movement stopper were exploded. The findings may be useful for cardiac valves designers in evaluating the impact of designing features of a valve on its hydrodynamic characteristics. PMID- 10475791 TI - [Interference testing in certification of medical equipment]. AB - The paper considers requirements for noise immunity of medical equipment, types of electromagnetic disturbances, and a list of medical apparatuses intended for obligatory certification testing for noise immunity. The paper presents experimental data on the impact of electromagnetic disturbances on different types of medical apparatuses. PMID- 10475792 TI - Ceiling fan injuries: the Townsville experience. PMID- 10475793 TI - [Effect of body temperature on changes in the blood Il-6 and IL-8 concentrations during open heart surgery]. PMID- 10475794 TI - [Study of the coating and reinforcing effect of Advaseal, a new synthetic sealant, on esophageal sutures]. PMID- 10475796 TI - Constrictive pericarditis. PMID- 10475795 TI - [Antitumor effect of a matrix metalloproteinase inhibitor, TA-2516 (Marimastat), on a nude mouse model of intraperitoneal seeding of stomach cancer]. PMID- 10475797 TI - Clinical outcomes after hepatitis C infection from contaminated anti-D immune globulin. PMID- 10475798 TI - Clinical outcomes after hepatitis C infection from contaminated anti-D immune globulin. PMID- 10475801 TI - Chronic myeloid leukemia. PMID- 10475799 TI - Rapidly fatal infection with Ehrlichia chaffeensis. PMID- 10475800 TI - A common B-cell precursor in composite lymphomas. PMID- 10475803 TI - The pathogenesis of melanoma induced by ultraviolet radiation. PMID- 10475802 TI - The pathogenesis of melanoma induced by ultraviolet radiation. PMID- 10475805 TI - Disease management. PMID- 10475804 TI - Disease management. PMID- 10475806 TI - Geriatrics and the limits of medicine. PMID- 10475808 TI - Lorazepam for the prevention of recurrent seizures related to alcohol. PMID- 10475809 TI - Lorazepam for the prevention of recurrent seizures related to alcohol. PMID- 10475811 TI - Subarachnoid hemorrhage in a patient with cerebral malaria. PMID- 10475810 TI - Smoking cessation. PMID- 10475807 TI - False diagnosis of maple syrup urine disease owing to ingestion of herbal tea. PMID- 10475812 TI - Preoperative pulmonary evaluation. PMID- 10475813 TI - Preoperative pulmonary evaluation. PMID- 10475814 TI - Preoperative pulmonary evaluation. PMID- 10475816 TI - Eating disorders. PMID- 10475815 TI - Eating disorders. PMID- 10475817 TI - Views of managed care. PMID- 10475818 TI - Views of managed care. PMID- 10475819 TI - Views of managed care. PMID- 10475820 TI - Views of managed care. PMID- 10475821 TI - Alopecia associated with indinavir therapy. PMID- 10475822 TI - Treatment of high-risk cervical cancer. PMID- 10475823 TI - Treatment of high-risk cervical cancer. PMID- 10475824 TI - Passive smoking and coronary heart disease. PMID- 10475825 TI - Passive smoking and coronary heart disease. PMID- 10475826 TI - Passive smoking and coronary heart disease. PMID- 10475827 TI - Passive smoking and coronary heart disease. PMID- 10475828 TI - Passive smoking and coronary heart disease. PMID- 10475829 TI - Passive smoking and coronary heart disease. PMID- 10475830 TI - Acute myocardial infarction after the use of sildenafil. PMID- 10475832 TI - Clinical features of neonates with hyperinsulinism. PMID- 10475834 TI - Educating others about issues surrounding end-of-life care. PMID- 10475833 TI - Managed care arrives in Latin America. PMID- 10475835 TI - ["Stop and train". Ambulatory lung training program in the Zurich central railway station]. PMID- 10475831 TI - Subaortic obstruction after sildenafil in a patient with hypertrophic cardiomyopathy. PMID- 10475836 TI - [Female mortality: lessons from geopathological comparisons]. AB - Worldwide a highly significant correlation exists between male and female mortality, particularly for all-cause, cardiovascular and cerebrovascular mortality, but to a lesser degree for cancer. This shows that the same factors are the major determinants of mortality in both sexes. Mortality from all causes over the age range 35-84 years can adequately be expressed by Gompertz equations (log mortality versus age), of which the intercepts are lower and the slopes higher in women compared to men. The demonstrates that ageing progresses more slowly in women. Maximum life expectancy is slightly higher in women. The various causes of mortality are competitive. PMID- 10475838 TI - Smallpox: setting the research agenda. PMID- 10475837 TI - Many of modes of transport for an embryo's signals. PMID- 10475839 TI - E-publication proposal. PMID- 10475840 TI - E-publication proposal. PMID- 10475841 TI - E-publication proposal. PMID- 10475842 TI - Chiron's licensing policy. PMID- 10475843 TI - Development of DNA sequencer. PMID- 10475845 TI - A new molecular window on early life. PMID- 10475844 TI - How chaperones protect virgin proteins. PMID- 10475846 TI - Deep Green rewrites evolutionary history of plants. PMID- 10475848 TI - Animal research. 50 monkeys taken from Indian lab. PMID- 10475847 TI - Biotechnology. New genes boost rice nutrients. PMID- 10475850 TI - The man who would spin genes into gold. PMID- 10475849 TI - Mining the genome for drugs. PMID- 10475851 TI - 6th Meeting on Toxicology of the French Society for the Study of Toxins and Symposium with the Society of Neurosciences. Paris, France, 9-11 December. Abstracts. PMID- 10475853 TI - Bioinformatics for all! ABRF meeting: Bioinformatics and Biomolecular Technologies: linking genomes, proteomes and biochemistry, Durham, North Carolina, USA, 19-22 March 1999. Association of Biomolecular Resource Facilities. PMID- 10475852 TI - Cerebral venous thrombosis: nothing, heparin, or local thrombolysis? PMID- 10475855 TI - The latest in signal transduction, 1999. Specificity in Signal Transduction, Keystone, Colorado, USA, 9-14 April 1999. PMID- 10475856 TI - The versatility of RNA structure and function. Jacques Monod Conference: New insights into the mechanism of mRNA translation: the significance of RNA structure, Aussois, France, 22-26 March 1999. PMID- 10475854 TI - Cutting in a lipid bilayer? Keystone Symposium on Molecular Mechanisms of Alzheimer Disease, Taos, New Mexico, USA, 3-8 March 1999. PMID- 10475857 TI - Age-related geriatric medicine or integrated medical care? PMID- 10475858 TI - The management of atrial fibrillation in elderly patients. PMID- 10475859 TI - Intensive care for very elderly patients: outcome and risk factors for in hospital mortality. AB - OBJECTIVES: To evaluate outcome and risk factors, particularly the Acute Physiology and Chronic Health Evaluation (APACHE) II scoring system, for in hospital mortality in very elderly patients after admission to an intensive care unit (ICU). METHODS: Retrospective chart review of patients > or =85 years admitted to the ICU. We recorded age, sex, previous medical history, primary diagnosis, date of admission and discharge or death, APACHE II score on admission, use of mechanical ventilation and inotropics, and complications during ICU admission. RESULTS: 104 patients > or =85 years (1.3% of all ICU admissions) were studied. The ICU and in-hospital mortality rates for these patients were 22 and 36% respectively. Factors correlated with a greater in-hospital mortality were: an admission diagnosis of acute respiratory failure (chi2; P = 0.007), the use of mechanical ventilation (chi2; P = 0.00005) and inotropes (chi2; P = 0.00001), complications during ICU admission (chi2; P = 0.004), in particular acute renal failure (chi2; P = 0.005), and an APACHE II score > or =25 (chi2; P = 0.001). The APACHE II scoring system and the use of inotropes were independently correlated with mortality. CONCLUSION: ICU and in-hospital mortality are higher in very elderly patients, particularly in those with an APACHE II score > or =25. The most important predictors of mortality are the use of inotropes and the severity of the acute illness. PMID- 10475861 TI - Why do so few older people with aortic stenosis have valve replacement surgery? AB - BACKGROUND: Many older patients with severe aortic stenosis do not have valve replacement surgery. OBJECTIVE: To determine the proportion of older people with symptomatic aortic stenosis referred for specialist assessment and the reasons for non-referral. METHODS: Retrospective study of all patients over 75 attending the geriatric department of an English teaching hospital. Confirmation was by reviewing all echocardiographic reports. RESULTS: Of 40 patients with symptomatic aortic stenosis, only four had undergone surgery. Seven patients refused operation; five were medically unfit (for reasons other than heart failure) and reasons for not referring could not be found in 13 casenotes. Of the 15 patients referred to a cardiologist, eight were considered suitable for surgery. CONCLUSION: Only one in five patients over 75 with echocardiographically confirmed aortic stenosis was considered for surgery. Prospective studies are needed to determine why patients and geriatricians do not seek specialist assessment for this treatable condition. PMID- 10475860 TI - The association of visual field deficits and visuo-spatial neglect in acute right hemisphere stroke patients. AB - BACKGROUND: Visuo-spatial neglect (VSN) after stroke is associated with a poor prognosis for rehabilitation. The co-existence of a visual field deficit (VFD) with VSN may be associated with impaired visuo-spatial functioning and thereby poor functional outcome. OBJECTIVE: To determine whether the presence of a VFD (i) exacerbates VSN and (ii) influences recovery of VSN. METHODS: A prospective study of consecutive acute (<7 days), right-hemisphere stroke patients who were able to undergo detailed assessment of visuo-spatial functioning and visual fields. Clinical assessment and a standardized neuropsychological test was administered by one observer, followed by independent assessment of visual fields by a second observer. Patients were followed up for 12 weeks with 4-weekly re assessments. RESULTS: 44 consecutive patients (23 women) with a first in a lifetime, acute hemisphere stroke were recruited. Twenty had VSN and VFD, seven VSN only, one VFD only and 17 had normal visual fields and no neglect. The finding of a VFD was significantly associated with the presence of VSN (P<0.0001). Patients with both VFD and VSN had a significantly lower score on the behavioural inattention test. One month post-stroke, this difference was no longer significant. Recovery of VSN and VFD was maximal in the first month, however VSN recovery continued for up to 12 weeks. Patients with VSN and a VFD on admission had a greater mortality at 1 and 3 months. CONCLUSION: The presence of a VFD does appear to exacerbate neglect in the acute stroke patient; this effect is no longer seen after 1 month. Recovery of VSN continues independent of a VFD. Patients with neglect and a VFD have an increased mortality, probably because of greater neurological impairment. PMID- 10475863 TI - The attitudes of patients to integrated medical care. AB - AIM: To establish the attitude of inpatients to integrated medical care. SETTING: A district general hospital which had recently adopted the model of integrated medical care. METHODS: 205 patients of all ages were interviewed and asked about their general views and their personal experience of integrated medical care and their preference had they been given a choice. Those patients who had previously been admitted before integration were asked if they felt the quality of care had changed. RESULTS: Integrated medical wards were preferred by most patients and this preference was greater in older patients. However, 30% identified patients on their wards who they felt should be segregated on account of confusion, frailty or youthfulness. Of the 69 who had been admitted before integration, 88% commented that the quality of care was either the same or better. CONCLUSION: Integrated medical care was the preferred model of care, especially by elderly patients. The special needs of some patients were, however, recognized. Perhaps the ideal model is a needs-related policy within a fully integrated system. PMID- 10475862 TI - Effect of immobilization on vitamin D status and bone mass in chronically hospitalized disabled stroke patients. AB - OBJECTIVE: To assess the influence of immobilization upon vitamin D status and bone mass in chronically hospitalized, disabled, elderly patients following stroke. DESIGN: cross-sectional study. SETTING: Department of geriatric neurology in a Japanese hospital. SUBJECTS: 129 chronically hospitalized, disabled, elderly stroke patients and 28 age-matched controls. RESULTS: We observed a deficiency of both 1,25-dihydroxyvitamin D (1,25-[OH]2D; 24.3 pg/ml) and 25-hydroxyvitamin D concentrations (25-OHD; 11.7 ng/ml) in stroke patients compared with controls. A high serum ionized calcium (mean; 2.648 mEq/l) was an independent determinant of the Barthel index (66) and 1,25-[OH]2D. When the patients were categorized into three groups by 25-OHD level (deficient, insufficient and sufficient), there was no difference in the mean 1,25-[OH]2D levels. Parathyroid hormone levels were normal or low and did not correlate with 25-OHD. Serum bone turnover variables and bone mineral density (BMD) of the second metacarpal in patients were significantly decreased compared to control subjects. Independent determinants of BMD included Barthel index, 25-OHD and 1,25-[OH]2D. CONCLUSIONS: 1,25-[OH]2D deficiency in immobilized stroke patients is not caused by substrate (25-OHD) deficiency but by hypercalcaemia. Immobilization-induced hypercalcaemia may inhibit parathyroid hormone secretion and thus 1,25-[OH]2D production, resulting in decreased BMD. Immobilization itself also may be responsible for decreased BMD. Exogenous 1,25-[OH]2D (calcitriol) rather than dietary vitamin D supplementation may be required in disabled elderly stroke patients who have a deficiency of 1,25-[OH]2D in order to prevent hip fractures, which frequently occur in this population. PMID- 10475864 TI - Goal attainment scaling as a measure of clinically important change in nursing home patients. AB - OBJECTIVES: To assess the feasibility, validity and responsiveness of an individualized measure--goal attainment scaling-in long-term care. DESIGN: Prospective descriptive study. SETTING: One academic and three community-based long-term care facilities. SUBJECTS: 53 nursing-home patients seen in consultation between July 1996 and June 1997. INTERVENTION: Specialized geriatric medicine consultation. MAIN OUTCOME MEASURES: Effect size and relative efficiency of the Barthel index, hierarchical assessment of balance and mobility, global deterioration scale, axis 8 (behaviour) of the brief cognitive rating scale, cumulative illness rating scale and the goal attainment scale. RESULTS: Mean goal attainment scale at follow-up was 46+/-7. The goal attainment scale was the most responsive measure, with an effect size of 1.29 and a relative efficiency of 53.7. The goal attainment scale did not correlate well with the other measures ( 0.22 to 0.17). CONCLUSION: Goal attainment scaling is a feasible and responsive measure in long-term care. Although fewer problems in nursing-home patients than elderly inpatients are susceptible to intervention, clinically important goals can be achieved in this population. PMID- 10475866 TI - Is there a relationship between balance, gait performance and muscular strength in 75-year-old women? AB - OBJECTIVE: To see if there is a relationship between clinical and laboratory tests of balance, muscular strength and gait in elderly women. DESIGN: A randomized population-based study. SETTINGS: Malmo, Sweden. METHODS: We investigated balance with a simple test of standing on one leg, as well as a computerized balance platform. Muscular strength was tested by computerized dynamometer. Extension and flexion of the knee and dorsiflexion of the ankle were tested. We measured the time and number of steps taken to walk a certain distance and the subjects' height and weight. PARTICIPANTS: 418 randomly selected 75-year old women, of whom 230 took part. RESULTS: There was no relation between the computerized balance tests and any of the other tests. The non-computerized balance test was correlated with gait time and number of steps (r = -0.50, P<0.001 and r = -0.40, P<0.001, respectively). Tests of extension and flexion, strength of the knee and ankle dorsiflexion were related to gait, speed and number of steps. Heavy women had poorer balance when assessed by the non computerized test (r = -0.32, P<0.001) and with the computerized, stable platform, eyes-open test (r = 0.27, P<0.001) and eyes-closed test (r = 0.44, P<0.001). The heavier an individual was, the slower her gait and the shorter her steps, despite having stronger knee muscles. CONCLUSION: There is no relationship between the simple balance tests and computerized platform tests. Muscle strength of the leg is not necessarily linked to balance, but rather to gait performance. PMID- 10475867 TI - The association between demographic factors, disease severity and the duration of symptoms at clinical presentation in elderly people with dementia. AB - OBJECTIVE: To determine the association between factors unrelated to the disease process, the duration of symptoms and the degree of cognitive or functional impairment in elderly patients presenting with dementia. METHOD: The living situation, educational level, age, gender and diagnosis based on standardized criteria were recorded for 209 elderly patients presenting to a memory clinic with dementia. Cognitive and functional deficits were measured with the cognitive section of the Cambridge Mental Disorders of the Elderly Examination combined with the Mini-Mental State Examination and the abbreviated version of the Blessed dementia scale, respectively. RESULTS: 129 patients had a diagnosis of probable Alzheimer's disease, 19 had probable ischaemic vascular dementia and 61 had mixed dementia. There was no effect of diagnosis on duration of symptoms or dementia severity at the time of presentation. Patients living with a son or daughter were more functionally impaired than those living alone or with a spouse. Males had higher cognitive scores but did not have milder functional deficits. Patients with only a primary-school education had a trend towards lower cognitive scores at presentation but did not have more functional deficits. CONCLUSIONS: The gender of the patient and the relationship to the carer are associated with cognitive and functional scores at the time of presentation in patients with dementia. PMID- 10475868 TI - Mortality in relation to urinary and faecal incontinence in elderly people living at home. AB - OBJECTIVE: To examine the relationship between incontinence and mortality in elderly people living at home. DESIGN: Of the randomly selected people aged 65 years and older living in Settsu city, Osaka in October 1992, 1405 were contacted and constituted the study cohort. Follow-up for 42 months was completed for 1318 (93.8%; 1129 alive, 189 dead). MEASURES: Data on general health status, history of health management, psychosocial conditions and urinary and faecal incontinence were collected by interview during home visits at the time of enrolment. RESULTS: From the Kaplan-Meier analysis, the estimated survival rates decreased with a decline in continence in both the 65-74 and 75 years and older age groups. From the Cox proportional hazards model, unadjusted hazard ratios of minor, moderate and severe incontinence for mortality, compared with continence, were 2.27, 2.96 and 5.94, respectively. Multivariate analysis yielded adjusted hazard ratios of minor, moderate and severe incontinence of 0.99, 1.17 and 1.91, respectively, leaving severe incontinence as the significant factor, when other indicators are controlled. CONCLUSIONS: Incontinence is related to mortality and severe incontinence represents an increased risk factor for mortality in elderly people living at home. PMID- 10475865 TI - Hand-grip strength predicts incident disability in non-disabled older men. AB - OBJECTIVES: To verify if hand-grip performance in older men is a predictor of disability. DESIGN: Population-based prospective study. SETTING: A sample from the Italian rural cohorts of the FINE study (Finland, Italy, Netherlands Elderly), representative of the general population of elderly men surveyed in 1991 and 1995. PARTICIPANTS: 140 men aged 71-91 years who reported no disability in performing activities of daily living (ADLs), instrumental activity of daily living (IADLs) and mobility activities at baseline examination and provided information on their functional status at follow-up 4 years later. MEASUREMENTS: Disability was defined as needing help in performing ADLs, IADLs and mobility. Hand-grip strength was evaluated at baseline by a mechanical dynamometer. RESULTS: After adjusting for potential confounding variables, a lower concentration of high-density lipoprotein cholesterol was the only factor predicting disability in men aged 76 years or younger and only reduced hand-grip strength predicted incident disability in men 77 years or older. CONCLUSION: Poor hand strength as measured by hand-grip is a predictor of disability in older people. The hand-grip test is an easy and inexpensive screening tool to identify elderly people at risk of disability. PMID- 10475869 TI - Carer distress in the general population: results from the Sydney Older Persons Study. AB - OBJECTIVE: To assess distress in a sample of carers who were selected from a community survey rather than recruited via community-service agencies. METHODS: A community survey was carried out on 630 people aged 75 or over living in Sydney, Australia. Informants nominated by these elderly people were divided into full carer (n = 21), partial carer (n = 187) and non-carer groups (n = 344). Informants completed the General Health Questionnaire (a continuous measure of psychiatric symptoms), the life satisfaction index (a measure of well-being) and the interpersonal bonding measure (a measure of quality of the relationship with the elderly person). Elderly participants had a medical examination, were assessed for disability and were questioned about use of services. RESULTS: Elderly people who had a full carer were more disabled and had more medical diagnoses. Full, but not partial, carers reported more psychiatric symptoms and lower life satisfaction. In multivariate analysis, the main determinant of carer distress was a relationship in which the carer felt controlled by the elderly person. CONCLUSION: When carers are selected from a population-based sample, only those who are full carers are more distressed. However, relationship factors are the most important determinant of distress. PMID- 10475870 TI - Cholesterol and long-term mortality after acute myocardial infarction in elderly patients. AB - METHOD: We investigated the association of total serum cholesterol concentrations and subsequent overall and coronary mortality in 304 patients aged > or =65 discharged from hospital after acute myocardial infarction. RESULTS: There was no association between total cholesterol concentrations and mortality due to either coronary heart disease or to all causes in all patients or, separately, in men, women, patients younger than 75 and patients aged 75 years and older. PMID- 10475871 TI - Patterns of self-reported health care use in injured and uninjured older adults. AB - METHOD: Using cross-sectional data obtained from the 1994 National Population Health Survey, we have compared self-reported health-care use between injured and non-injured older adults. RESULTS: Over 12 months, 9% of non-institutionalized people over 65 reported an injury. The commonest injuries were sprains or strains and fractures and half of these were caused by a fall. Older people sustaining injuries are more likely to spend a night in hospital and receive physiotherapy. PMID- 10475872 TI - Appendix abscess: a surgical giant presenting as a geriatric giant. AB - CASE REPORT: A women aged 102 years presented with falls and was found to have an atypical presentation of appendicitis. CONCLUSION: This illustrates the non specific presentation of disease in old age and the importance of a careful medical assessment of people who have fallen. PMID- 10475873 TI - Cervical osteoarthropathy: an unusual cause of dysphagia. AB - PRESENTATION: A 72-year-old man complained of progressive dysphagia for solids associated with a sensation of foreign body in his throat for 2 years. A barium swallow showed a bridging osteophyte between C4 and C5 vertebrae indenting the oesophagus posteriorly and displacing it anteriorly. OUTCOME: He refused surgical intervention and was given dietary advice. After 6 months, his weight was steady and he was able to swallow semi-solid food without difficulty. PMID- 10475874 TI - 'Ikigai' in older Japanese people. PMID- 10475875 TI - Drug-induced agranulocytosis in older people. A case series of 25 patients. PMID- 10475877 TI - Rectal examinations in elderly subjects. PMID- 10475876 TI - Do we overlook respiratory symptoms and airflow obstruction in elderly medical admissions? PMID- 10475880 TI - Driving safety: motivating messages. PMID- 10475878 TI - Inter-rater reliability of the clock-drawing test. PMID- 10475879 TI - Heart failure: a diagnostic and therapeutic dilemma in elderly patients. PMID- 10475881 TI - Too many congresses? PMID- 10475882 TI - Clinical polymorphism of respiratory allergy. PMID- 10475883 TI - The water depression flowmeter. PMID- 10475886 TI - Prospective evaluation of risk factors for bloodstream infection in patients receiving home infusion therapy. AB - BACKGROUND: Intravenous therapy in the outpatient and home settings is commonplace for many diseases and nutritional disorders. Few data are available on the rate of and risk factors for bloodstream infection among patients receiving such therapy. OBJECTIVE: To determine rates of and risk factors for bloodstream infection among patients receiving home infusion therapy. DESIGN: Prospective, observational cohort study. SETTING: Cleveland, Ohio, and Toronto, Ontario, Canada. PATIENTS: Patients receiving home infusion therapy through a central or midline catheter. MEASUREMENTS: Primary laboratory-confirmed bloodstream infection. RESULTS: Among 827 patients (988 catheters), the most common diagnoses were infections other than HIV (67%), cancer (24%), nutritional and digestive disease (17%), heart disease (14%), receipt of bone marrow or solid organ transplants (11%), and HIV infection (7%). Sixty-nine bloodstream infections occurred during 69,532 catheter-days (0.99 infections per 1000 days). In a Cox regression model with time-dependent covariates, independent risk factors for bloodstream infection were recent receipt of a bone marrow transplant (hazard ratio, 5.8 [95% CI, 3.0 to 11.3]), receipt of total parenteral nutrition (hazard ratio, 4.1 [CI, 2.3 to 7.2]), receipt of therapy outside the home (for example, in an outpatient clinic or physician's office) (hazard ratio, 3.6 [CI, 2.2 to 5.9]), use of a multilumen catheter (hazard ratio, 2.8 [CI, 1.7 to 4.7]), and previous bloodstream infection (hazard ratio, 2.5 [CI, 1.5 to 4.2]). Rates of bloodstream infection per 1000 catheter-days varied from 0.16 for patients with none of these 5 risk factors to 6.77 for patients with 3 or more risk factors. Centrally inserted venous catheters were associated with a higher risk than implanted ports were, but the difference was not statistically significant. CONCLUSION: Bloodstream infections seem to be infrequent among outpatients receiving infusions through central and midline catheters. However, the rate of infection increases with bone marrow transplantation, parenteral nutrition, infusion therapy in a hospital clinic or physician's office, and use of multilumen catheters. Compared with implanted ports or peripherally inserted catheters, centrally inserted venous catheters may confer greater risk for bloodstream infection. PMID- 10475885 TI - Serum total homocysteine concentrations in the third National Health and Nutrition Examination Survey (1991-1994): population reference ranges and contribution of vitamin status to high serum concentrations. AB - BACKGROUND: The concentration of circulating total homocysteine is a sensitive marker of inadequate folate and vitamin B12 status. Elevated homocysteine concentrations are associated with an increased risk for vascular disease. OBJECTIVE: To identify reference ranges for serum total homocysteine concentration in U.S. residents and quantify the contribution of circulating vitamin concentrations to high homocysteine concentrations. DESIGN: Cross sectional prevalence study. SETTING: United States. PATIENTS: A nationally representative sample of 3563 male participants and 4523 female participants 12 years of age or older who participated in the third National Health and Nutrition Examination Survey. MEASUREMENTS: Reference ranges (5th and 95th percentiles) for the total homocysteine concentration were defined among participants who were folate- and vitamin B12-replete and had normal creatinine concentrations. A high total homocysteine concentration was defined as one that exceeded the sex specific 95th percentile for the reference sample (participants 20 to 39 years of age). The population attributable risk percentage was calculated to determine the contribution of low folate (<11 nmol/L) and vitamin B12 (<185 pmol/L) concentrations to a high homocysteine concentration. RESULTS: Reference ranges for serum total homocysteine concentration increased with age; these ranges were 4.3 to 9.9 micromol/L for male participants and 3.3 to 7.2 micromol/L for female participants 12 to 19 years of age and from 5.9 to 15.3 micromol/L for men and 4.9 to 11.6 micromol/L for women 60 years of age or older. A high homocysteine concentration was defined as at least 11.4 micromol/L for male participants and at least 10.4 micromol/L for female participants. Approximately two thirds of the cases of high homocysteine concentrations were associated with low vitamin concentrations. CONCLUSIONS: Upper reference limits for the serum total homocysteine concentration increased with age and were higher for male participants than for female participants at all ages. In most cases, high homocysteine concentrations were associated with low serum vitamin concentrations. PMID- 10475884 TI - Nonfasting plasma total homocysteine level and mortality in middle-aged and elderly men and women in Jerusalem. AB - BACKGROUND: Elevated plasma total homocysteine level has been associated with cardiovascular disease in many studies, mostly in Europe and North America. Data on persons from other areas and on associations with overall mortality are sparse. OBJECTIVE: To determine the relation of plasma homocysteine level to all cause and cause-specific mortality. DESIGN: Prospective observational study with 9- to 11-year follow-up. SETTING: A free-living, multiethnic Jewish population in western Jerusalem, Israel. PARTICIPANTS: 1788 residents of Jerusalem (808 men and 980 women) who were at least 50 years of age and were examined between 1985 and 1987 as part of the Kiryat Yovel Community Health Study. MEASUREMENTS: Nonfasting plasma homocysteine level was determined in frozen stored samples. Deaths during follow-up were identified by linkage with the national population registry. RESULTS: Plasma homocysteine levels exceeded 14 micromol/L in 28% of men and 20% of women. During the study period, 405 deaths occurred. In multivariate Cox models that controlled for possible confounders, a nonmonotonic increase in mortality hazard ratios was associated with ascending quintile of homocysteine level: 1.0, 1.4, 1.3, 1.5, and 2.0 (P < 0.001 for trend). The relation was similar for cardiovascular and noncardiovascular causes of death (excluding cancer). The association was weaker when deaths that occurred during the first 5 years of follow-up were excluded; corresponding hazard ratios for ascending quintile of homocysteine level were 1.0, 1.0, 1.2, 1.1, and 1.6 (P = 0.063 for trend). Age- and sex-adjusted percentages of deaths "attributable" to elevated plasma homocysteine level (> or = 14 micromol/L) were 12.5% (95% CI, 6.7% to 18.8%) for all deaths, 16.0% (CI, 7.2% to 25.6%) for deaths during the first 5 years of follow-up, and 8.3% (CI, 1.5% to 16.1%) for later deaths. CONCLUSIONS: A mildly to moderately elevated nonfasting plasma homocysteine level is a substantial risk marker for death from all causes. The association seems to be stronger during the first 5 years of follow-up. PMID- 10475887 TI - Normalization of hyperhomocysteinemia with L-thyroxine in hypothyroidism. AB - BACKGROUND: Hyperhomocysteinemia is an independent risk factor for coronary, peripheral, and cerebrovascular disease. Elevated plasma homocysteine levels were described in a preliminary report on primary hypothyroidism. OBJECTIVE: To determine whether restoration of euthyroidism by L-thyroxine replacement therapy would reduce or normalize plasma homocysteine levels. DESIGN: Prospective cohort study. SETTING: Outpatient endocrinology department of a tertiary center. PATIENTS: 14 patients (10 women and 4 men; 25 to 77 years of age): 4 with newly diagnosed chronic (Hashimoto) hypothyroidism and 10 who had been rendered acutely hypothyroid (thyroid-stimulating hormone level > 25 mU/L) by total thyroidectomy for thyroid carcinoma. MEASUREMENTS: Total plasma homocysteine levels were measured at baseline and 3 to 9 months later, after euthyroidism had been attained by L-thyroxine replacement therapy. RESULTS: Median baseline plasma homocysteine levels in both sexes (women, 11.65 micromol/L [range, 7.2 to 26.5 micromol/L]; men, 15.1 micromol/L [range, 14.1 to 16.3 micromol/L]) were higher (P = 0.002) than those in healthy female (n = 35) and male (n = 36) volunteers (women, 7.52 micromol/L [range, 4.3 to 14.0 micromol/L]; men, 8.72 micromol/L [range, 5.94 to 14.98 micromol/L]). Eight patients (57%) had baseline plasma homocysteine levels that exceeded the upper limit of sex-specific reference ranges. Upon attainment of euthyroidism, all patients had a diminution in plasma homocysteine levels. The median overall change of -5.5 micromol/L (range, -15.4 to -1.8 micromol/L) corresponds to a difference of -44% (range, -58% to -13%) (P < 0.001). Homocysteine levels returned to normal in 7 of the 8 patients with elevated pretreatment values. CONCLUSIONS: Hypothyroidism may be a treatable cause of hyperhomocysteinemia, and elevated plasma homocysteine levels may be an independent risk factor for the accelerated atherosclerosis seen in primary hypothyroidism. PMID- 10475889 TI - Update in cardiology. PMID- 10475890 TI - Homocyst(e)ine and cardiovascular disease: a critical review of the epidemiologic evidence. AB - PURPOSE: To review epidemiologic studies on the association between homocyst(e)ine level and risk for cardiovascular disease and the potential benefits of homocysteine-decreasing therapies. DATA SOURCES: Computerized and manual searches of the literature on total homocysteine levels and cardiovascular disease. STUDY SELECTION: Prospective studies and major retrospective epidemiologic studies evaluating the association between homocyst(e)ine levels and cardiovascular disease and the association between blood levels or dietary intake of folate, vitamin B6, and vitamin B12 and cardiovascular disease. DATA EXTRACTION: Relevant data on patient population, plasma homocyst(e)ine levels, duration of follow-up, and main results were extracted from studies that met the inclusion criteria. DATA SYNTHESIS: The designs and results of studies included in this review are summarized. A formal meta-analysis was not performed because the studies were heterogeneous in method and design. CONCLUSIONS: Results of epidemiologic studies suggest that moderately elevated plasma or serum homocyst(e)ine levels are prevalent in the general population and are associated with an increased risk for cardiovascular disease, independent of classic cardiovascular risk factors. Simple, inexpensive, nontoxic therapy with folic acid, vitamin B6, and vitamin B12 reduces plasma homocyst(e)ine levels. Although the association between homocyst(e)ine levels and cardiovascular disease is generally strong and biologically plausible, the data from the prospective studies are less consistent. In addition, epidemiologic observations of an association between hyperhomocyst(e)inemia and cardiovascular risk do not prove the existence of a causal relation. Therefore, the effectiveness of folate, vitamin B6, and vitamin B12 in reducing cardiovascular morbidity and mortality requires rigorous testing in randomized clinical trials. Several such trials are under way; their results may greatly affect cardiovascular morbidity and mortality, given the simplicity and low cost of vitamin therapy. PMID- 10475891 TI - Potential new cardiovascular risk factors: left ventricular hypertrophy, homocysteine, lipoprotein(a), triglycerides, oxidative stress, and fibrinogen. AB - The 1996 Bethesda Conference acknowledged left ventricular hypertrophy, hyperhomocysteinemia, lipoprotein(a) excess, hypertriglyceridemia, oxidative stress, and hyperfibrinogenemia as possible new cardiac risk factors. This review summarizes the current literature that supports these conditions as cardiac risk factors. Left ventricular hypertrophy is an independent risk factor for vascular disease. Improvement or progression of left ventricular hypertrophy influences subsequent cardiovascular complications. Clinical trials are under way to assess the potential benefit of decreasing homocysteine levels. The role of lipoprotein(a) excess in vascular disease is controversial. The atherogenic potential of lipoprotein(a) seems to be neutralized by effective reduction of low density lipoprotein cholesterol levels. Increasing evidence supports an independent role of hypertriglyceridemia in cardiovascular disease and a possible clinical benefit from decreasing triglyceride levels. Among antioxidant micronutrients, supplementation with vitamin E has been shown to be beneficial in primary and secondary prevention studies. Data supporting the use of other antioxidants are much weaker. Preliminary evidence suggests that reducing fibrinogen levels in patients with high baseline levels and coronary disease may be beneficial. Despite the potential relation between new risk factors and cardiovascular disease, routine clinical application of these conditions as cardiovascular risk factors would be premature. Evidence is needed that these conditions extend prognostic ability beyond conventional risk factors and that modification of these conditions can reduce the risk for cardiovascular events. PMID- 10475892 TI - Homocysteine in health and disease. PMID- 10475893 TI - Medical Writings revisited. PMID- 10475888 TI - Nonfasting plasma total homocysteine levels and stroke incidence in elderly persons: the Framingham Study. AB - BACKGROUND: Total homocysteine levels are associated with arteriosclerotic outcomes. OBJECTIVE: To determine whether total homocysteine levels predict incident stroke in elderly persons. DESIGN: Prospective population-based cohort study with 9.9 years of follow-up. SETTING: Framingham, Massachusetts. PATIENTS: 1947 Framingham Study participants (1158 women and 789 men; mean age +/- SD, 70 +/- 7 years). MEASUREMENTS: Baseline total homocysteine levels and 9.9-year stroke incidence. RESULTS: The quartiles of nonfasting total homocysteine levels were as follows: quartile 1, 4.13 to 9.25 micromol/L; quartile 2, 9.26 to 11.43 micromol/L; quartile 3, 11.44 to 14.23 micromol/L; quartile 4, 14.24 to 219.84 micromol/L. During follow-up, 165 incident strokes occurred. In proportional hazards models adjusted for age, sex, systolic blood pressure, diabetes, smoking, and history of atrial fibrillation and coronary heart disease, relative risk (RR) estimates comparing quartile 1 with the other three quartiles were as follows: quartile 2 compared with quartile 1--RR, 1.32 (95% CI, 0.81 to 2.14); quartile 3 compared with quartile 1--RR, 1.44 (CI, 0.89 to 2.34); quartile 4 compared with quartile 1--RR, 1.82 (CI, 1.14 to 2.91). The linear trend across the quartiles was significant (P < 0.001). CONCLUSION: Nonfasting total homocysteine levels are an independent risk factor for incident stroke in elderly persons. PMID- 10475894 TI - Dietary vitamin A intake and risk for hip fracture. PMID- 10475895 TI - Vulnerable plaque. PMID- 10475897 TI - Vulnerable plaque. PMID- 10475898 TI - Decrease in endothelin-1 plasma levels during the menstrual cycle and after ethinylestradiol treatment. PMID- 10475896 TI - Vulnerable plaque. PMID- 10475899 TI - Centennial physician: Richard Perry, MD, struggles with the future. PMID- 10475900 TI - Experimental and modeling studies of diffusion in immobilized cell systems. A review of recent literature and patents. PMID- 10475901 TI - [Treatment of the chronic form of Chagas disease. Is the etiologic treatment efficient?]. PMID- 10475902 TI - Minor labial salivary glands. Proceedings of a satellite symposium to the XVth Jubilee Arkovy Congress of Hungarian Dentists. Budapest, Hungary, August 28, 1998. PMID- 10475903 TI - Rapid solid phase extraction cleanup for pesticide residues in fresh fruits and vegetables. PMID- 10475904 TI - A one-step ASE extraction method for TCDD TEQ determination. PMID- 10475905 TI - Suitability of high-hardness COMBO medium for ecotoxicity testing using algae, daphnids, and fish. PMID- 10475906 TI - Significant organotin contamination of sediment and tissues of milkfish in brackish water ponds. PMID- 10475907 TI - Heavy metal pollution in water, sediments, and earthworms from the Ebro River, Spain. PMID- 10475908 TI - Surface water transport of lead at a shooting range. PMID- 10475909 TI - Mercury levels in mole crabs Hippa cubensis, Emerita brasiliensis, E. portoricensis, and Lepidopa richmondi (Crustacea: Decapoda: Hippidae) from a sandy beach at Venezuela. PMID- 10475910 TI - Selenium and copper in vegetables and fruits grown on long-term impacted soils from Valparaiso region, Chile. PMID- 10475911 TI - Biodegradation of polyvinylchloride (PVC) by white rot fungi. PMID- 10475912 TI - Plasma aspartate aminotransferase activity in the catfish Clarias albopunctatus exposed to sublethal zinc and mercury. PMID- 10475914 TI - Effects of ammonium nitrate, sodium nitrate, and urea on red-legged frogs, Pacific treefrogs, and African clawed frogs. PMID- 10475915 TI - Acute lethal and sublethal effects of a neem-based insecticide on nontarget aquatic insects in stream channels. PMID- 10475913 TI - Acute-sublethal copper effects on phagocytosis and lysozyme activity in the earthworm Amynthas hawayanus. PMID- 10475916 TI - Coexposure to epichlorohydrin on the elimination of urinary metabolites of dimethylformamide. PMID- 10475917 TI - Effect of sublethal concentrations of monocrotophos on erythropoietic activity and certain hematological parameters of fish Anabas testudineus (Bloch). PMID- 10475918 TI - Acute toxicity of carbofuran to adult and juvenile flathead chubs. PMID- 10475919 TI - Growth differences in cultured populations of Brachionus plicatilis Muller caused by heavy metal stress as function of microalgal diet. PMID- 10475920 TI - Identification of ammonia and volatile phenols as primary toxicants in a coal gasification effluent. PMID- 10475921 TI - Estimation of the hazard concentration of industrial wastewaters using algal bioassay. PMID- 10475923 TI - Radiologic evaluation of dysphagia. PMID- 10475925 TI - Three-dimensional intraluminal sonography in the evaluation of gastrointestinal diseases. PMID- 10475924 TI - Three-dimensional endoluminal ultrasound: a new method for the evaluation of gastrointestinal tumors. AB - BACKGROUND: The purpose of the present study was to evaluate the feasibility of three-dimensional endoluminal ultrasound of gastrointestinal tumors. METHODS: Sixteen patients with esophageal, gastric, or colorectal tumors underwent endoscopic ultrasound. Three-dimensional ultrasound data were obtained from multiple serial images of a miniprobe (360 degrees, 12.5 MHz) and processed on a PC-based 3D workstation. RESULTS: Adequate three-dimensional ultrasound scans were obtained in eight patients with esophageal cancer and five patients with colorectal cancer. Three-dimensional image processing enabled visualization of the data as a multiplanar display or as a life-like three-dimensional view. The availability of arbitrary scan planes improved the assessment of local tumor spread and the spatial relation of the tumor to relevant adjacent structures (e.g., major vessels). Three-dimensional presentations provided realistic views of the anatomy and facilitated the interpretation of the ultrasound images. CONCLUSIONS: Three-dimensional display and the ability to review endoluminal ultrasound data interactively may improve the staging of gastrointestinal tumors. These preliminary data encourage further evaluation of this technique. PMID- 10475926 TI - Acute diverticulitis of the small bowel: CT findings. AB - We present the computed tomographic (CT) findings in two cases of small bowel diverticulitis, one affecting the jejunum and the other a Meckel's diverticulum. The main CT finding was that of a mass with an air-fluid collection in contiguity with small bowel loops. PMID- 10475927 TI - Diffusion-weighted MR imaging with single-shot echo-planar imaging in the upper abdomen: preliminary clinical experience in 61 patients. AB - BACKGROUND: To determine the potential ability of diffusion-weighted magnetic resonance (MR) imaging with single-shot echo-planar imaging (DW imaging) in the upper abdomen by apparent diffusion coefficient (ADC) and signal:intensity ratio (SIR) measurements. METHODS: DW imaging was performed in 61 clinical patients. ADCs in the liver, pancreas, spleen, kidney, and different pathological conditions were calculated. Spleen-to-liver SIR and segmental intensity difference of the liver (SID) were also calculated. RESULTS: The mean ADCs (mm(2)/s) were 2.28 x 10(-3) +/- 0.07 in the liver, 1.44 x 10(-3) +/- 0.05 in the spleen, 1.94 x 10(-3) +/- 0.19 in the pancreas, and 5.76 x 10(-3) +/- 0.06 in the kidney. The mean ADC of cirrhotic liver was 1.96 x 10(-3) +/- 0.62, which was lower than that of normal liver. Other pathologic conditions also showed ADCs different from those of normal tissues. All DW images showed significantly higher spleen-to-liver SIRs and SIDs than did T2-weighted images (p < 0.05). CONCLUSION: The mean ADCs obtained with DW imaging were different in each upper abdominal organ and with each pathologic condition. DW images showed better soft tissue contrast than did T2-weighted images with regard to SIR and CNR in depicting and characterizing upper abdominal disorders. PMID- 10475928 TI - Necrotic granuloma of the liver by human fascioliasis: imaging findings. AB - We report a case of necrotic granuloma of the liver by human fascioliasis. The lesion showed unusual findings at sonography, computed tomography, and magnetic resonance imaging that have not been reported previously. PMID- 10475922 TI - The oblique pharyngogram: value in the assessment of dysphagia. AB - BACKGROUND: To evaluate the diagnostic value of oblique views of the pharynx in patients with dysphagia. METHODS: One hundred thirty-three patients with symptoms referable to the cervical region underwent pharyngoesophography that included views of the pharyngoesophageal junction filmed at three frames per second and spot films of the pharynx obtained in distended frontal, lateral, and both oblique projections. Examination was completed with assessment of the entire esophagus and gastric cardia. RESULTS: The oblique views identified abnormalities not shown on the standard views in 5% of patients. The oblique views proved useful in 12%, where the lower pharynx was obscured in the lateral projection by large shoulders, and in 18% to assess the valleculae when this region was obscured by the occiput and mandible in the frontal projection. In 12%, the oblique views proved useful in demonstrating normal structures when artifacts raised the possibility of lesions on the standard projections. In 10%, poor technique impaired visualization of pharyngeal structures on the standard projections, but repeat swallows in the oblique projections proved adequate to assess these regions. Three (27%) of the 11 cases of cervical esophageal webs were best seen on oblique views, and in another three patients the webs were visible only on oblique views. CONCLUSIONS: Oblique views are of value in the assessment of the pharynx. There will be instances when the standard projections are inadequate, and these alternative views will complement the evaluation of this region. The addition of oblique views will sometimes improve the confidence of the interpretation of normality or assist evaluation of the extent of an abnormality. PMID- 10475929 TI - Hepatic imaging studies on patients with visceral larva migrans due to probable Ascaris suum infection. AB - Visceral larva migrans (VLM) is a disease usually observed in children in which the larvae of animal parasites invade and reside in human tissues for long periods. Although the common causal species of VLM are Toxocara canis and T. cati, we identified three adult patients with VLM, probably due to Ascaris suum, whose diagnosis was made by specific immunoserological tests. The patients complained of respiratory symptoms, and laboratory tests showed pronounced eosinophilia, but neither larvae nor eggs were detected in stool samples. We present the findings of various imaging studies of the patients. Multiple small hypoechoic mass lesions were demonstrated by ultrasound tomography, which disappeared after anti-helminthic therapy. Hepatic mass lesions were detected as low-density areas on computed tomography, as high signal intensities on T2 weighted magnetic resonance images, as space-occupying regions in liver scintigraphy, and as yellow-white nodules in laparoscopy. Although biopsied liver tissue specimens showed marked infiltrations of eosinophiles in the portal tracts and hepatic sinusoids, neither larvae nor eggs could be identified. PMID- 10475932 TI - Multinodular fatty change in the liver in patients with chronic hepatic porphyria. AB - We present five cases of chronic hepatic porphyria (CHP) without skin lesions, in whom a multinodular fatty change in the liver was incidentally detected by ultrasound. Discovery prompted further examinations, leading to the diagnosis of CHP. Although relatively rare, the possibility of CHP must be considered when encountering patients with irregular (multinodular) fatty infiltration of the liver of unknown cause. We also discuss the differential diagnosis. PMID- 10475930 TI - Biliary ascariasis: CT, MR cholangiopancreatography, and navigator endoscopic appearance--report of a case of acute biliary obstruction. AB - A case of acute common bile duct obstruction caused by a roundworm in a 6-year old girl is reported. Computed tomography (CT) and magnetic resonance imaging (MRI) showed "bull's-eye" and "eye-glass" appearances of the ascaris in the common bile duct. On reformation of the transverse CT and coronal MR images, the tubular Ascaris was better depicted. MRI and navigator endoscopic demonstrations of the common bile duct ascaris have not been described previously in the literature. These CT and MRI findings may be helpful in the diagnosis of Ascaris. PMID- 10475931 TI - Optimal phases of dynamic CT for detecting hepatocellular carcinoma: evaluation of unenhanced and triple-phase images. AB - BACKGROUND: To determine the optimal phases of dynamic computed tomography (CT) for detecting hepatocellular carcinoma (HCC). METHODS: Fifty-two patients with 85 HCC nodules were examined by means of unenhanced and triple-phase CT images of the whole liver. The time for obtaining the arterial-phase images was 25-55 s after intravenous bolus injection of contrast material, the time for obtaining the portal venous-phase images was 65-100 s, and the time for obtaining late phase images was 145 s to 4 min. Detectability of the HCC nodules for all phases was statistically compared. RESULTS: The detection rates for the arterial- and late-phase images were significantly higher than for the unenhanced and portal venous-phase images (p < 0.01). The combination of arterial- and late-phase images showed the same number of HCC nodules in the same number of patients as did the combination of unenhanced and triple-phase images. CONCLUSION: The combination of the arterial- and late-phase imagings was best for detecting HCC nodules. PMID- 10475933 TI - Prediction of the histologic type of bile duct cancer by using intraductal ultrasonography. AB - BACKGROUND: Patients with papillary adenocarcinoma survive longer than do patients with other histologic types of bile duct tumors. We evaluated the usefulness of intraductal ultrasonography (IDUS) for predicting the histology. METHODS: Preoperative tumor assessment was performed by using IDUS through a percutaneous tract or the transpapillary route in 37 patients with extrahepatic bile duct cancer. In 30 of 37 patients, imaging results were compared prospectively with histologic findings in resected specimens. Probes 2.0 mm in diameter and 20 MHz in frequency were mainly used. When IDUS showed a "narrow based polypoid pattern" or a "papillary surface pattern," the patients were judged as having papillary adenocarcinoma. RESULTS: The accuracy, sensitivity, and specificity of IDUS in predicting papillary adenocarcinoma were 90%, 89%, and 90%, respectively. When intraductal ultrasonography showed a papillary surface pattern or a narrow-based polypoid pattern, lymph node metastases and perineural invasion were rarely seen when compared with other patients with bile duct cancer (p < 0.05). CONCLUSION: IDUS is useful for assessing the histologic type of bile duct cancer. PMID- 10475934 TI - Spontaneous thrombosis of a pseudoaneurysm complicating pancreatitis. AB - Patients with a visceral aneurysm are at high risk for acute transpapillary, intra-, or retroperitoneal hemorrhage, necessitating either surgical or endovascular therapy. We report an instance of spontaneous thrombosis of a pseudoaneurysm complicating pancreatitis before endovascular treatment could be performed. Causality and the literature of spontaneous thrombosis in pseudoaneurysms are discussed. PMID- 10475935 TI - Rupture of a pancreatic pseudocyst into the portal venous system. AB - Pseudocyst formation is a well-known complication of acute or chronic pancreatitis. We report a case in which pseudocyst ruptured into the splenic and portal veins. PMID- 10475936 TI - Hydrocolonic sonography in the evaluation of colonic lesions. AB - BACKGROUND: The present study was done to assess the sensitivity, specificity, ease of examination, and limitations of hydrocolonic sonography (HCS) to evaluate a heterogeneous spectrum of colonic disorders. METHODS: In a prospective study, 100 patients (53 male, 47 female) with clinical suspicion of colonic abnormality were examined by conventional abdominal sonography, HCS, and colonoscopy on the same day. The patients then underwent appropriately planned barium studies. Histopathologic diagnosis was established by colonoscopic/excision biopsies or fine needle aspiration cytology (FNAC) whenever indicated and results correlated. RESULTS: In 94% of patients, it was possible to evaluate the entire colon from the rectosigmoid junction to the cecum. Redundant parts of the colon were not well evaluated by this technique. The rectum could not be adequately evaluated, and rectal examination was not included in the study. HCS can evaluate in great detail both the colonic lumen and wall and, hence, could suggest the nature of the lesion in the majority (87.9%) of cases based on characteristic changes in wall stratification, echogenicity, lumenal changes, and site and length of involvement, although distinguishing between benign and malignant lesions was not always possible. In 93% of patients with tuberculous colitis/ileocolitis, the normal wall stratification was no longer in evidence, with moderate hypoechoic thickening of the wall (average = 8.34 mm). Malignant lesions (93.7%) showed grossly thickened bowel wall with loss of stratification and hypoechoic/heterogeneous echo texture. Intralumenal polypoid masses also were seen in 87.5% of cases, and there was extension beyond the adventitia and involvement of pericolonic tissues in 75% of cases. All patients with ulcerative colitis had only mild hypoechoic wall thickening; the five-layer structure could be clearly discerned in 87.5% of cases. HCS had an overall sensitivity of 90.9% and a specificity of 94.7% in this study. CONCLUSIONS: HCS can be advantageously used for diagnosis and differential diagnosis of inflammatory and malignant colonic lesions and also for follow-up of patients with chronic inflammatory large bowel diseases such as ulcerative colitis. PMID- 10475937 TI - Hydrocolonic sonography. PMID- 10475938 TI - Disseminated peritoneal plasmacytoma following cardiac transplantation. AB - Posttransplant lymphoproliferative disorders (PTLD) are an important comorbidity in immunosuppressed transplant patients. We describe a unique case of PTLD, a disseminated peritoneal plasmacytoma in a 56-year-old cardiac transplant patient presenting with ascites. The associated computed tomographic findings include omental thickening, mesenteric stranding, and ascites. PMID- 10475939 TI - Adrenal endothelial cyst with massive central calcification. AB - A 55-year-old woman was treated for a rare case of adrenal endothelial cyst that was detected incidentally during management of abdominal pain. Computed tomography of the tumor displayed massive central calcification and a fluid-fluid level by intracystic hemorrhage. Magnetic resonance imaging showed an intracystic papillary part with calcification. After surgery, the tumor was diagnosed as an angiomatous type of adrenal endothelial cyst. PMID- 10475940 TI - Cervical carcinoma: efficacy of thin-section oblique axial T2-weighted images for evaluating parametrial invasion. AB - BACKGROUND: To investigate the efficacy of thin-section oblique axial T2-weighted images in the assessment of parametrial invasion by cervical carcinoma. METHODS: One hundred parametria of 50 patients with cervical carcinoma were evaluated with pathologic correlation. We compared the sensitivity, specificity, and diagnostic accuracy in the assessment of parametrial invasion by cervical carcinoma between axial T2-weighted images and thin-section oblique axial T2-weighted images. RESULTS: Thin-section oblique axial T2-weighted images provided accurate cross sections of the cervix with excellent detail and detected parametrial invasion more accurately than did axial T2-weighted images showing cross sections of the trunk. Although the sensitivity, specificity, and accuracy for parametrial invasion were 46.4%, 91.7%, and 79.0%, respectively, on axial T2-weighted images, the corresponding values were 67.9%, 97. 2%, and 89.0%, respectively, on thin section oblique axial T2-weighted images. There were statistically significant differences in the sensitivity (p = 0.014), specificity (p = 0.046), and accuracy (p = 0.002) in detecting parametrial invasion between these two types of images. CONCLUSIONS: Thin-section oblique axial T2-weighted images are useful for the assessment of parametrial invasion by cervical carcinoma. PMID- 10475941 TI - Endometrial carcinoma: efficacy of thin-section oblique axial MR images for evaluating cervical invasion. AB - BACKGROUND: To investigate the efficacy of thin-section oblique axial magnetic resonance (MR) images in evaluating cervical invasion by endometrial carcinoma. METHODS: Sixty-seven patients with endometrial carcinoma were evaluated with pathologic correlation. We compared the accuracy in the assessment of cervical invasion by endometrial carcinoma between parasagittal MR images and thin-section oblique axial MR images by using T2-weighted and contrast-enhanced T1-weighted pulse sequences. RESULTS: Cervical invasion by endometrial carcinoma was confirmed by pathologic examination. Cervical invasion was seen in 16 patients. The accuracy rates of parasagittal T2-weighted images, thin-section oblique axial T2-weighted images, parasagittal contrast-enhanced T1-weighted images, and thin section oblique axial contrast-enhanced T1-weighted images were 74.7%, 89.5%, 82.0%, and 95.5%, respectively. Statistically significant differences were seen between parasagittal T2-weighted images and thin-section oblique axial T2 weighted images (p = 0.002) and between parasagittal contrast-enhanced T1 weighted images and thin-section oblique axial contrast-enhanced T1-weighted images (p = 0.003). CONCLUSION: Thin-section oblique axial MR images are considered to be useful for the assessment of cervical invasion by endometrial carcinoma. PMID- 10475944 TI - Venous ulcer reappraisal: insights from an international task force. Veines International Task Force. AB - An international task force made up of a panel of 16 experts was mandated to review and objectively evaluate all aspects of chronic venous disease of the leg. All available publications on chronic venous disease of the leg from 1983 to 1997 were identified through computerized search. Three different screenings were then performed in order to select only relevant papers providing moderate to strong scientific evidence. Final conclusions and further therapeutic recommendations were made based on these publications. Compression, medications, local therapies, sclerotherapy and surgery are the existing therapeutic options for which the following recommendations can be made. Compression: Properly applied bandages both fixed and stretched have been shown to be effective. Compression stockings may be used. Compression needs to be in excess of 35 mm Hg. Medications: Although preliminary results have shown a beneficial effect of several vasodilators and oral micronized purified flavonoid fraction, the evidence for the efficacy of medications on venous ulcer healing is still limited and further studies are required before recommendations can be made. There is no evidence to routinely administer antibiotics. Local therapies have an as yet unproven adjunctive role. Sclerotherapy is unlikely to be effective unless there is superficial venous insufficiency, in which it may have an as yet unproven role. SURGERY: Patients with active venous ulcer and sapheno-femoral or sapheno-popliteal junction incompetence benefit from surgical treatment. In the absence of reflux or following deep vein thrombosis, there is no evidence to support surgical treatment. PMID- 10475945 TI - 1996-97 Global Anthrax Report. AB - While there is a general decrease in the number of anthrax outbreaks, and thus of human cases, worldwide this is still a disease that is extensively under diagnosed and under-reported. However, it is now very infrequent to rare in Canada, the United States, and many countries in Europe. An increasing number of countries are now free. At the other extreme, it is a significant problem in West Africa, Spain, Greece, Turkey, Albania, Romania and in Central Asia. In spite of the textbooks, livestock and wildlife deaths do occur, sometimes commonly, without any 'diagnostic' extravasation of blood and, if not realised, infected carcasses get recycled into meat and bone meals for feed. PMID- 10475946 TI - A national register of historic and contemporary anthrax foci. AB - Anthrax in Russia has for a long time posed a serious problem for public health and veterinary services. At the beginning of the century, 40-60 thousand cases of this infection were annually reported in the country in agricultural animals and about 10-20 thousand cases in people where each fourth (25%) was dying. In the Russian Federation the registration of anthrax foci is obligatory for veterinary as well as for sanitary-epidemiological services. So our initial project, funded by the International Technical and Scientific Center in Moscow, has envisaged the collection of all data of known anthrax foci, including the village name, agricultural council, region, oblast and year of occurrence. The objective is to assemble a reference handbook, "Register of stable anthrax sites in the Russian Federation", containing organized information on more than 10 000 anthrax foci occurring during the past 100 years. Such a study makes it possible to identify regions characterized by the highest concentrations of stationary anthrax sites in Russia, to identify trends in expressed activity of such sites through the periodic emergence of disease in humans and animals, and to determine the factors contributing to the formation of such trends. In doing this, it makes it possible to develop contingency plans for different risk locations (i.e. high risk of persistent infection, high risk of sporadic occurrence, low risk areas, etc.) in terms of anthrax in Russia, to identify high risk areas and develop a differentiated strategy of vaccination and other control strategies, and to develop preventive recommendations to reduce risk in high risk areas. It is now important to develop the second step of the project: to develop in depth studies of particular clusters to determine which factors are associated with Russian anthrax outbreaks. Maps will greatly enhance the value of this work in terms of spatial analysis. Furthermore, this supplementary project will allow the incorporation of powerful Geographic Information System (GIS) electronic mapping technology so that natural geographic features, such as soil type, climate, etc., can be compared with anthrax distributions in Russia using standard GIS and statistical analysis. At the present time, it is imperative to develop a detailed understanding of the world's distribution of anthrax and what geographic factors determine its prevalence. PMID- 10475947 TI - Anthrax explodes in an Australian summer. AB - Anthrax occurred on 83 properties in an area of north central Victoria between 26 January and 26 March in the summer of 1997. Anthrax had not been recorded in the outbreak area since records were initiated in 1914, although anthrax did occur in the general area in the 1880s to 1890s. Standard Australian control measures were applied to the properties, including quarantine, tracing movements of animals on and off affected properties, secure disposal of carcases by burning, enhanced surveillance of stock generally in the area and the use of local disaster control procedures including an alert of health authorities. As affected property numbers began to increase dramatically from 8 February, it was decided to use blanket area vaccination to control the disease. By 26 February, the epidemic curve had returned to the base line and a buffer vaccination zone of 457 farms holding 78,649 cattle was formed by early March 1997. Between 26 January and 26 March when the outbreak was declared over, 202 cattle and 4 sheep were confirmed to have died of anthrax. Between 27 March and early November a further 26 cattle were confirmed as dying due to anthrax and 14 of these had not had previous vaccination, including four young calves and one horse. One new property within the vaccination buffer zone had an anthrax case in a cow in early November 1997. By mid-November 1997, all previously infected and all neighbouring properties within 1 km were compulsorily re-vaccinated, as were all calves when two months of age and all introduced cattle. In 1998, only two confirmed cases of anthrax were diagnosed; both were vaccinated calves on farms which had had multiple cases during the outbreak. The public reaction and attention fueled by unprecedented media attention led to intense international scrutiny from countries where anthrax is a particular zoonotic problem. Very strong representations had to be made about the safety of livestock and livestock products that came from Victoria. This event has demonstrated that there is a need to review OIE and other requirements and recommendations covering anthrax where strict restrictions are placed on livestock and livestock products to protect livestock and human populations against anthrax infection. PMID- 10475948 TI - Identification of Bacillus anthracis strains in China. AB - As part of a project to establish a reference strain collection, phenotypic, biochemical and genetic analyses were carried out on 84 strains of Bacillus anthracis, 81 of them of Chinese origin from various sources. Particular differences from reports on isolates of other origins were the possession of fimbriae and single polar flagella with consequent motility in 77, self agglutination by 64 and failure to ferment maltose in 60 of the Chinese strains. The findings were considered to be of significance to identification, classification and evolution of B. anthracis. PMID- 10475949 TI - Meso-scale ecology of anthrax in southern Africa: a pilot study of diversity and clustering. AB - It has only recently been possible to detect sufficient genetic diversity among anthrax isolates to allow genotype grouping (Keim et al. 1997). Early results of such grouping suggest that the southern African subcontinent may be the geographical origin of Bacillus anthracis. This report describes a pilot investigation of the genetic diversity of a study group of isolates from the Kruger National Park, South Africa, and efforts to detect spatio-temporal clustering within the study group. This study has also served as further validation for the newly developed Multi-Locus VNTR Analysis (MLVA), designed to simplify genotyping of B. anthracis isolates. The results reveal a diverse range of genotypes within the park allied with three genotype reference groups, and show that the MLVA procedure is robust for rapid analysis of B. anthracis genotypes. We also observed multiple genotype groups within epidemics and between geographically and temporally close epidemic episodes. This is in contrast to earlier characterizations of anthrax epidemics. The result of a Mantel test for time-space clustering indicates clustering of the anthrax isolates selected for the study. PMID- 10475950 TI - A review of anthrax in Canada and implications for research on the disease in northern bison. AB - During the first half of the century, the majority of anthrax outbreaks in Canada occurred in the southern portions of Ontario and Quebec and were often associated with pastures contaminated by effluent from textile industries dealing with imported animal materials. In 1952, introduction of Federal regulations requiring disinfection of these materials greatly reduced the incidence of anthrax in eastern Canada. Since 1962, domestic outbreaks of the disease have been reported almost exclusively in cattle in the western prairie provinces. Between 1962 & 1993, nine anthrax epizootics have been recorded in the bison herds of the Northwest Territories and northern Alberta resulting in the deaths of at least 1309 animals. During the northern epizootics there has been a strong sex bias in mortalities with the majority of carcasses being sexually mature bulls. The northern epizootics occur during drought conditions in the late summer, preceded by a wet spring, and end with the arrival of coolers weather. It has been hypothesized that stress factors associated with these meteorological conditions coupled with breeding stress during the late summer rut may predispose the bulls to infection. Alternatively, the meteorological conditions may work to concentrate anthrax spores in the environment into low lying wallows preferentially utilized by the bulls. Recent genetic analyses of Bacillus anthracis isolates from Canada and the United States have identified that, while closely related to isolates from domestic outbreaks, isolates from northern bison epizootics form their own distinct strain. This suggests that the establishment of anthrax in northern Canada was a singular event that occurred prior to the first recognized epizootic in 1962. A review of the agricultural history of northern Canada has identified several situations in the first half of the century which may have provided the opportunity for the transfer of anthrax from cattle to the indigenous bison. PMID- 10475951 TI - Antibody-based systems for the detection of bacillus anthracis in environmental samples AB - There is a necessity for rapid immunodiagnostic techniques for the detection and identification of Bacillus anthracis in environmental specimens. The technology available for accomplishing this ranges in complexity from a simple dipstick type assay to complex biosensors. We have developed antigen capture dipstick assays for a series of infectious agents including an assay for B. anthracis protective antigen and one for B. anthracis spores. These immunochromatographic assays use colloidal gold to visualize the reaction and take approximately 15 min to perform. We will also describe our current effort in the development of two antigen detection biosensors and discuss the sensitivity and specificity of the assays in environmental specimens. PMID- 10475952 TI - Molecular diversity in Bacillus anthracis. AB - Molecular typing of Bacillus anthracis has been extremely difficult due to the lack of polymorphic DNA markers. We have identified nine novel variable number tandemly repeated loci from previously known amplified fragment length polymorphism markers or from the DNA sequence. In combination with the previously known vrrA locus, these markers provide discrimination power to genetically characterize B. anthracis isolates. The variable number tandem repeat (VNTR) loci are found in both gene coding (genic) and non-coding (non-genic) regions. The genic differences are 'in frame' and result in additions or deletion of amino acids to the predicted proteins. Due the rarity of molecular differences, the VNTR changes represent a significant portion of the genetic variation found within B. anthracis. This variation could represent an important adaptive mechanism. Marker similarity and differences among diverse isolates have identified seven major diversity groups that may represent the only world-wide B. anthracis clones. The lineages reconstructed using these data may reflect the dispersal and evolution of this pathogen. PMID- 10475953 TI - Fluorescent detection techniques for real-time multiplex strand specific detection of Bacillus anthracis using rapid PCR. AB - Speed is a key area in our development of PCR assays for Bacillus anthracis. We believe that the strand specific detection of amplicons within 10 min is a realistic goal and that this will be achieved through fluorescent in-tube assays. We have used the Idaho LightCycler to study and develop candidate assays for B. anthracis. New strand specific fluorescent methods have been developed and a number of formats have been studied for speed and sensitivity. Internal controls have been developed as a method of improving our assay confidence. In this communication we will introduce the field of rapid PCR whilst discussing previous work in the areas described above, the development of our own rapid assay and a novel internal control system for B. anthracis. This work used PCR assays and hardware that are either commercially available, or have been previously described in open literature publications. PMID- 10475954 TI - The Ba813 chromosomal DNA sequence effectively traces the whole Bacillus anthracis community. AB - Plasmid genes that are responsible for virulence of Bacillus anthracis are important targets for the DNA-based detection of anthrax. We evaluated the distribution of the Ba813 chromosomal DNA sequence (Ba813) within closely related Bacillus species. Ba813 was systematically identified from 47 strains or isolates of B. anthracis tested, thus indicating its reliability as a tracer for that species. From the 60 strains of closely related Bacillus spp. examined, three bona fide B. cereus and one bona fide B. thuringiensis were found to harbour Ba813. This marker was also detected in Bacillus sp. isolates that were present at high levels in soil samples collected in a place where an anthrax outbreak had occurred. The significance and the possible function of the Ba813 locus is discussed. PMID- 10475955 TI - Polymerase chain reaction-ELISA to detect Bacillus anthracis from soil samples limitations of present published primers. AB - A polymerase chain reaction (PCR)-ELISA technique to detect Bacillus anthracis from soil samples has been developed. The application of streptavidine-coated microtitre plates as well as plates covered with covalently linked oligonucleotides as catching probes led to a test sensitivity of about 100 fg pure genomic DNA or of less than 10 spores seeded into 100 g soil material. Some non-suspicious soil samples collected from different locations yielded positive results with presently published primers or probes targeting the B or C gene of pX02 and with primers targeting the chromosomal sequence B813. The former PCR products were sequenced. The number and mode of base exchanges led to the assumption that there will exist at least one unknown soil organism with high similarity within highly conserved capsule-encoding genes. PMID- 10475956 TI - Definitive identification of Bacillus anthracis--a review. AB - The word 'problem' is seen with some frequency in relation to clear differentiation between Bacillus anthracis and B. cereus. In fact, although the close relationship of these two species is undisputed, it is only in the case of a few borderline isolates, rarely encountered in practice, that any sort of identification problem exists. Until recently this was only important to the taxonomist who found it unsatisfactory not to be able to identify definitively such isolates. To most others, if the isolate was unable to produce anthrax in a laboratory animal, it was discarded as irrelevant without being named, or it was called B. cereus or given a name such as B. anthracis similis, or even a totally unrelated name. More recently, in view of the new light in which B. anthracis is increasingly seen, resulting from its putative association with bioaggression, clear identification has become a more critical issue. This paper reviews the current state of the art and suggests the way forward for the future. PMID- 10475957 TI - Characterization of the exosporium of Bacillus cereus. AB - Exosporium components from endospores of Bacillus cereus ATCC 10876 were purified and separated by gel electrophoresis. Several of the proteins for which N terminal sequences were recovered were found to have homologues in protein databases which have been demonstrated to have enzymic activity in other organisms. Amongst these is a zinc metalloprotease, immune inhibitor A, already described in B. thuringiensis. This has been shown to be present in an active 73 kDa form on the exosporium of B. cereus. Other proteins associated with the exosporium include the molecular chaperone GroEL and a homologue of RocA (1 pyrroline-5-carboxylate dehydrogenase (EC 1.5.1.12)) of B. subtilis. Although these are unlikely to represent integral structural proteins of the exosporium, the observation that they are selectively present in the spore surface layer suggests that this layer may have functional significance. PMID- 10475959 TI - The capsule of bacillus anthracis, a review AB - The capsule of Bacillus anthracis, composed of poly-D-glutamic acid, serves as one of the principal virulence factors during anthrax infection. By virtue of its negative charge, the capsule is purported to inhibit host defence through inhibition of phagocytosis of the vegetative cells by macrophages. In conjunction with lethal toxin and oedema toxin, whose target cells include macrophages and neutrophils, respectively, the capsule allows virulent anthrax bacilli to grow virtually unimpeded in the infected host. Spores germinating in the presence of serum and elevated CO2 release capsule through openings on the spore surface in the form of blebs which may coalesce before sloughing of the exosporium and outgrowth of the fully encapsulated vegetative cell. It has not been established that spore encapsulation plays a role in the early events of anthrax infection. The capsule appears exterior to the S-layer of the vegetative cell and does not require the S-layer for its attachment to the cell surface. The three membrane associated enzymes required for synthesis of the capsule are encoded by the 60 MDa pX02 plasmid. The cistrons are arranged in the order capB, capC and capA and encode for proteins of 44, 16 and 46 kDa, respectively. The synthesis of capsule and toxin is, in part, under bicarbonate regulation by interaction of transacting proteins of the atxA gene on the 100-MDa pX01 toxin-encoding plasmid and the acpA gene on the pX02 plasmid. Therefore, capsule synthesis is enhanced in the presence of the atxA gene on the pX01 plasmid. An additional protein (with a predicted size of 51 kDa) is encoded by the dep gene located downstream from the cap region and appears to be a depolymerase that catalyses the hydrolysis of poly D-glutamic acid into lower molecular weight polyglutamates. Although the biological function of the Dep protein is unknown, it has been proposed that the low molecular weight polyglutamates produced by the action of the enzyme may act to inhibit host defence mechanisms. PMID- 10475958 TI - Molecular recognition specificity of Bacillus anthracis spore antibodies. AB - The sensitivity and specificity of polyclonal and monoclonal antibodies raised against anthrax spore preparations has been assessed by Western blotting. None of the antibodies studied were completely specific in recognizing the anthrax spore surface. A polyclonal serum recognized a wide range of spore surface epitopes and demonstrated limited cross-reaction with the near-neighbour species Bacillus cereus spore surface. Two monoclonal antibodies studied demonstrated more extensive cross-reaction with distant-neighbour species B. globigii and B. subtilis. These monoclonal antibodies did not react with spore surface epitopes but did bind strongly to vegetative cell epitopes in all four Bacillus species studied. PMID- 10475960 TI - Bacillus anthracis surface: capsule and S-layer. AB - Two abundant surface proteins, EA1 and Sap, are components of the Bacillus anthracis surface layer (S-layer). Their corresponding genes have been cloned, shown to be clustered on the chromosome and sequenced. EA1 and Sap each possess three 'S-layer homology' motifs. Single and double disrupted mutants were constructed. EA1 and Sap were co-localized at the cell surface of both the non capsulated and capsulated bacilli. When present, the capsule is exterior to, and completely covers, the S-layer proteins, which form an array beneath it. Nevertheless, the presence of these proteins is not required for normal capsulation of the bacilli. Thus both structures are compatible, and yet neither is required for the correct formation of the other. Bacillus anthracis has, therefore, a very complex cell wall organization for a gram-positive bacterium. PMID- 10475961 TI - The S-layer homology domain as a means for anchoring heterologous proteins on the cell surface of Bacillus anthracis. AB - Bacillus anthracis synthesizes two S-layer proteins, each containing three S layer homology (SLH) motifs towards their amino-terminus. In vitro experiments suggested that the three motifs of each protein were organized as a structural domain sufficient to bind purified cell walls. Chimeric genes encoding the SLH domains fused to the levansucrase of Bacillus subtilis were constructed and integrated on the chromosome. Cell fractionation and electron microscopy studies showed that both heterologous polypeptides were targeted to the cell surface. In addition, surface-exposed levansucrase retained its enzymatic and antigenic properties. Preliminary results concerning applications of this work are presented. PMID- 10475962 TI - Sequence, assembly and analysis of pX01 and pX02. AB - Bacillus anthracis plasmids pX01 and pX02, harboured by the Sterne and Pasteur strains, respectively, have been sequenced by random 'shotgun' cloning and high throughout sequence analysis. These sequences have been assembled (Sequencher) to generate a circulate pX01 plasmid containing 181 656 bp and a single linear (gapped) pX02 contig containing at least 93.479 bp. Initial annotation suggests that the two plasmids combined contain at least 200 potential open reading frames (ORFs) with < 40% having significant similarity to sequences registered in open databases. Collectively, only 118 566 bp of the pX01 DNA (65%) represent predicted coding regions. This value is similar to published gene densities for other plasmids and is indicative of the larger intergenic spaces in plasmids vs those found in the chromosomes of the parental microbes (85-93% gene density). A 70 kbp region including the toxin genes (cya, lef and pag) is distinct from the remainder of the pX01 sequence: (1) it has a lower gene density (58 vs 70%) than the remaining 111 kbp; (2) it contains all but one of the co-regulated transcriptional fusions identified by transposon mutagenesis (Hoffmaster & Koehler 1997) and (3) it contains a significantly higher proportion of positive BLAST scores (62 vs 20%) for putative ORFs. These data suggest different origins for the two regions of pX01. PMID- 10475963 TI - Genetic comparison of Bacillus anthracis and its close relatives using amplified fragment length polymorphism and polymerase chain reaction analysis. AB - Amplified fragment length polymorphism (AFLP) analysis allows a rapid, relatively simple analysis of a large portion of a microbial genome, providing information about the species and its phylogenetic relationship to other microbes (Vos et al. 1995). The method simply surveys the genome for length and sequence polymorphisms. The AFLP pattern identified can be used for comparison to the genomes of other species. Unlike other methods, it does not rely on analysis of a single genetic locus that may bias the interpretation of results and does not require any prior knowledge of the targeted organism. Moreover, a standard set of reagents can be applied to any species without using species-specific information or molecular probes. We are using AFLP analysis to rapidly identify different bacterial species. A comparison of AFLP profiles generated from a large battery of Bacillus anthracis strains shows very little variability among different isolates (Keim et al. 1997). By contrast, there is a significant difference between AFLP profiles generated for any B. anthracis strain and even the most closely related Bacillus species. Sufficient variability is apparent among all known microbial species to allow phylogenetic analysis based on large numbers of genetically unlinked loci. These striking differences among AFLP profiles allow unambiguous identification of previously identified species and phylogenetic placement of newly characterized isolates relative to known species based on a large number of independent genetic loci. Data generated thus far show that the method provides phylogenetic analyses that are consistent with other widely accepted phylogenetic methods. However, AFLP analysis provides a more detailed analysis of the targets and samples a much larger portion of the genome. Consequently, it provides an inexpensive, rapid means of characterizing microbial isolates to further differentiate among strains and closely related microbial species. Such information cannot be rapidly generated by other means. AFLP sample analysis quickly generates a very large amount of molecular information about microbial genomes. However, this information cannot be analysed rapidly using manual methods. We are developing a large archive of electronic AFLP signatures that is being used to identify isolates collected from medical, veterinary, forensic and environmental samples. We are also developing the computational packages necessary to rapidly and unambiguously analyse the AFLP profiles and conduct a phylogenetic comparison of these data relative to information already in our database. We will use this archive and the associated algorithms to determine the species identity of previously uncharacterized isolates and place them phylogenetically relative to other microbes based on their AFLP signatures. This study provides significant new information about microbes with environmental, veterinary and medical significance. This information can be used in further studies to understand the relationships among these species and the factors that distinguish them from one another. It should also allow the identification of unique factors that contribute to important microbial traits, including pathogenicity and virulence. We are also using AFLP data to identify, isolate and sequence DNA fragments that are unique to particular microbial species and strains. The fragment patterns and sequence information provide insights into the complexity and organization of bacterial genomes relative to one another. They also provide the information necessary for the development of species-specific polymerase chain reaction primers that can be used to interrogate complex samples for the presence of B. anthracis, other microbial pathogens or their remnants. PMID- 10475964 TI - The native virulence plasmid combination affects the segregational stability of a theta-replicating shuttle vector in Bacillus anthracis var. New Hampshire. AB - The segregational stability of a small, theta-replicating, non-mobilizable shuttle plasmid (pAEX-5E) was determined in fully virulent (pX01+/pX02+), partially cured (pX01+/pX02- and pX01-/pX02+) and fully cured (pX01-/pX02-) derivatives of Bacillus anthracis var. New Hampshire. Under the growth conditions used (L-broth, 37 degrees C, aerobic, batch culture), pAEX-5E remained segregationally stable in the pX01-/pX02+ and pX01-/pX02- derivatives for in excess of 100 culture generations, but was expelled from the pX01+/pX02+ and pX01+/pX02- derivatives (100% loss occurred after 101+/-3.8 and 54+/-6.0 culture generations, respectively). In the presence of antibiotic selection pressure to maintain pAEX-5E (5 microg erythromycin ml-1) no comparable loss of pX01 or pX02 was observed over 100 generations of growth in any of the derivatives of B. anthracis. Under these conditions the pX01+/pX02- derivative had an extended culture doubling time (td+/-S. E. of the mean) of 75.3 +/- 1.4 min compared with 47.3 +/- 1.1, 46.2 +/- 0.86 and 43.2 +/- 1.2 min for the pX01+/pX02+, pX01-/pX02+ and pX01-/pX02- derivatives, respectively. That antibiotic resistance was pAEX-5E mediated was confirmed using a second antibiotic marker (kanamycin). After100 generations of growth in the presence of erythromycin, colonies were shown to have retained kanamycin resistance. Southern blot analysis, in conjunction with plasmid rescue to Escherichia coli confirmed that, after 100 culture generations in the presence of antibiotic selection pressure, pAEX-5E had remained structurally stable and had not integrated into the B. anthracis genome. PMID- 10475965 TI - Control of virulence gene expression in Bacillus anthracis. AB - The atxA gene is an important regulator of virulence gene expression in Bacillus anthracis. atxA positively regulates expression of the three genes encoding the anthrax toxin proteins and at least one gene is required for capsule production. Here we report that an atxA-null mutant exhibits phenotypes unrelated to toxin and capsule synthesis. An atxA-null mutant grows poorly on minimal media and sporulates more efficiently than the parent strain. Numerous transposon-generated promoter-lacZ fusions at distinct loci on pXO1 exhibit CO2-enhanced atxA dependent expression similar to that observed for the toxin genes. We also report that the atxA-activated pagA gene (encoding the protective antigen toxin protein) is co-transcribed with a 300-bp gene, pagR, located downstream of pagA. The predicted protein product of pagR has some amino acid sequence similarity to transcriptional regulators in other organisms. Our data indicate that pagR represses expression of pagA and atxA. pagR also controls expression of some CO2/atxA-activated transcriptional fusions on pXO1 that do not correspond to the toxin genes. Regulation of these fusions and pagA and pagR may be due to changes in AtxA levels, or may be independent of atxA expression. PMID- 10475966 TI - Crystallographic studies of the anthrax lethal toxin AB - Anthrax lethal toxin comprises two proteins: protective antigen (PA; MW 83 kDa) and lethal factor (LF; MW 87 kDa). We have recently determined the crystal structure of the 735-residue PA in its monomeric and heptameric forms (Petosa et al. 1997). It bears no resemblance to other bacterial toxins of known three dimensional structure, and defines a new structural class which includes homologous toxins from other Gram-positive bacteria. We have proposed a model of membrane insertion in which the water-soluble heptamer undergoes a substantial pH induced conformational change involving the creation of a 14-stranded beta barrel. Recent work by Collier's group (Benson et al. 1998) lends strong support to our model of membrane insertion. 'Lethal factor' is the catalytic component of anthrax lethal toxin. It binds to the surface of the cell-bound PA heptamer and, following endocytosis and acidification of the endosome, translocates to the cytosol. We have made substantial progress towards an atomic resolution crystal structure of LF. Progress towards a structure of the 7:7 translocation complex between the PA heptamer and LF will also be discussed. PMID- 10475967 TI - Mechanism of membrane translocation by anthrax toxin: insertion and pore formation by protective antigen AB - Proteolytic activation of receptor-bound protective antigen (PA) at the cell surface removes PA20, allowing PA63 to oligomerize and form a ring-shaped heptameric prepore. The prepore binds edema factor (EF) and lethal factor (LF) and, after endocytosis and trafficking of the complex to an acidic, vesicular compartment, it undergoes membrane insertion and mediates translocation of EF/LF to the cytosol. Data from membrane conductance experiments support a model of membrane insertion in which the 2beta2-2beta3 loop of PA, which is disordered in native PA and the prepore, forms a 14-stranded transmembrane beta-barrel. Recent studies on the process of prepore-to-pore conversion and our current understanding of the mechanism of pH-dependent translocation will be described. PMID- 10475968 TI - Anthrax toxin fusion proteins for intracellular delivery of macromolecules AB - The dominant role played by the anthrax toxin in Bacillus anthracis pathogenesis shows that the toxin has evolved to be an efficient system for delivering its two catalytic protein components, oedema factor and lethal factor (LF), into the cytosol of host cells. This system involves binding of the protective antigen (PA) toxin component to a ubiquitous (and still unidentified) receptor, proteolytic activation at the cell surface, internalization by endocytosis and translocation through an early endosome membrane to the cytosol (Leppla 1995). We and colleagues showed that the system can be exploited to deliver heterologous polypeptides to the cytosol (Arora et al. 1992; Milne et al. 1995). This work used the catalytic domains of other toxins which are normally translocated across membranes (Arora & Leppla 1994). Immunity to intracellular pathogens depends on the cytosolic processing of antigens to produce peptides that are presented on the cell surface bound to MHC Class I molecules. The anthrax toxin delivery system provides a way to mimic this process. We made a fusion protein containing the (non-catalytic) amino terminal domain of LF and the gp120 envelope glycoprotein of HIV-1. Administration of this recombinant protein along with PA to antigen-presenting cells sensitized them to cytolysis by cytotoxic T-cells specific to gp120 peptides (Goletz et al. 1997). Further exploitation of the anthrax toxin system as a cell-targeting reagent would be facilitated by achieving cell type specificity. The recent determination of the PA structure (Petosa et al. 1997) allows rational engineering to modify or replace the receptor-binding domain with specific ligand structures. A model system was produced by fusing a c-Myc peptide to the carboxyl terminus of PA so as to target hybridoma cells expressing cell surface antibodies to this peptide. Killing of the hybridoma cells was shown to be specific by competition with the peptide and with non-toxic mutants of PA (Varughese et al. 1998). PMID- 10475969 TI - Lethal toxin actions and their consequences. AB - After entry of infectious anthrax spores into the body, host-specific signals induce spore germination, outgrowth of vegetative bacilli and the expression of lethal toxin and other virulence factors. Anthrax lethal toxin (LeTx) is a virulence factor responsible for the major pathologies seen during systemic anthrax infections. Injection of sterile LeTx into test animals mimics the shock and sudden death seen during active bacterial infections. Once large levels of LeTx are produced within the body, destruction of bacteria by administration of antibiotics is usually unsuccessful. The LeTx is believed to be secreted into the bloodstream where it circulates freely throughout the body and binds and enters host cells. Once in the cytoplasm, the lethal factor acts as a zinc metalloprotease disrupting normal homoeostatic functions. Macrophages are a uniquely sensitive cell type that seem to be vital global mediators of toxin induced pathologies. Removal of macrophages from mice renders them insensitive to LeTx challenge. Low levels of lethal toxin induce macrophage production, in vitro, of the shock-inducing cytokines TNF and Il-1beta. Higher levels of LeTx cause over-production of reactive oxygen intermediates, bursting of macrophages and release of mediators of shock. We believe that agents capable of blocking key steps of the lethal toxin cascade may prove useful in combating anthrax pathologies. PMID- 10475970 TI - Anthrax lethal factor cleaves the N-terminus of MAPKKS and induces tyrosine/threonine phosphorylation of MAPKS in cultured macrophages AB - The lethal toxin (LeTx) of Bacillus anthracis is the major virulence factor responsible for the death of infected animals and for cytolysis of cultured macrophages. Its catalytic component, LF, contains the characteristic zinc binding motif of metalloproteases, it binds zinc and indirect evidence suggests that this hydrolytic activity is essential for LeTx cytotoxicity (Limpel et al. 1994; Kochi et al. 1994). To identify substrates of LF, we have used the yeast two-hybrid system, employing an LF inactive mutant as bait. This approach has led to the identification of the MAP kinase kinases (MAPKKs) Mek1 and Mek2 as proteins capable of specific interaction with LF. LF cleaves Mek1 and Mek2 within their N-terminus in vitro and in vivo, hydrolysing a Pro8-Ile9 and a Pro10-Arg11 peptide bond in Mek1 and Mek2, respectively (Vitale et al. 1998), similarly to that found with a different approach by Duesbery et al. (1998). The removal of the amino terminus of MAPKKs eliminates the 'docking site' involved in the specific interaction with MAPKs and interferes with the phospho-activation of the MAPKs ERK1 and ERK2, which become phosphorylated in cultured macrophages following toxin challenge. We are currently investigating the relevance of MAPKKs cleavage for LeTx cytotoxicity and the consequences for the activity of the MAP pathway. PMID- 10475971 TI - Anthrax lethal factor causes proteolytic inactivation of mitogen-activated protein kinase kinase. AB - A search of the National Cancer Institute's Anti-Neoplastic Drug Screen for compounds with an inhibitory profile similar to that of the mitogen-activated protein kinase kinase (MAPKK) inhibitor PD098059 yielded anthrax lethal toxin. Anthrax lethal factor was found to inhibit progesterone-induced meiotic maturation of frog oocytes by preventing the phosphorylation and activation of mitogen-activated protein kinase (MAPK). Similarly, lethal toxin prevented the activation of MAPK in serum stimulated, ras-transformed NIH3T3 cells. In vitro analyses using recombinant proteins indicated that lethal factor proteolytically modified the NH2-terminus of both MAPKK1 and 2, rendering them inactive and hence incapable of activating MAPK. The consequences of this inactivation upon meiosis and transformed cells are also discussed. PMID- 10475972 TI - Experiences with vaccination and epidemiological investigations on an anthrax outbreak in Australia in 1997. AB - Between January and February 1997, there was a severe outbreak of anthrax on 83 properties in north-central Victoria, Australia. Vaccination was used as a major tool to control the outbreak by establishing a vaccination buffer zone 30 km by 20 km. In all, 78, 649 cattle in 457 herds were vaccinated in a three week program. In the face of the outbreak, there was a delay before vaccination was able to stop deaths. In the 10 days following vaccination 144 cases of confirmed anthrax occurred and 38 cases occurred more than 10 days after vaccination. When all cattle on at-risk properties were revaccinated in October and early November 1997, there were only two confirmed cases of anthrax in vaccinated seven and nine month old calves in the following anthrax season. Investigations into the epidemiology of the outbreak were unable to establish a single major association for the spread of the disease by flies, biting insects, carrion scavengers, wind, manufactured feed, milk factory tanker routes, veterinary visits, animal treatments, movements of personnel between farms or burning of carcases. The weather conditions in the outbreak area were part of a long dry spell with periods of high daily and night temperatures, continuing high humidity over the period and higher than normal soil temperatures. It is possible that extensive earth works in the district involving irrigated pasture renovation and water channel and drainage renovation could have disturbed old anthrax graves. It is postulated that these works released spores that were dispersed in the preceding wet winter across poorly drained areas that formed the axis for the outbreak. The earth moving renovations establishing irrigation in the area were conducted in the late 1890s, and before the occurrence of anthrax outbreaks were recorded. The axis of the outbreak was the major stock route for cattle and sheep moving from southern Victoria to northern Victoria and southern New South Wales, and undoubtedly there would have been extensive anthrax outbreaks before vaccine became available in the 1890s. In respect of other outbreaks, the events in Victoria most resembled outbreaks of anthrax recorded in the United States of America in the 1950s, 1960s and 1970s. PMID- 10475973 TI - Antigen delivery by attenuated Bacillus anthracis: new prospects in veterinary vaccines. AB - This report summarizes the recent investigations on the use of Bacillus anthracis as a live vector for delivery of antigens. Recombinant strains were constructed by engineering the current live Sterne vaccine. This vaccine, used to prevent anthrax in cattle, causes side-effects due to anthrax toxin activities. Bacteria producing a genetically detoxified toxin factor were devoid of lethal effects and were as protective as the Sterne strain against experimental anthrax. Moreover, B. anthracis expressing a foreign antigen controlled by an in vivo inducible promoter were able to generate either antibody or cellular protective responses against heterologous diseases. PMID- 10475974 TI - Clinical aspects, diagnosis and treatment of anthrax AB - There are three clinical presentations of anthrax in humans: cutaneous (>95% of cases), orogastric and inhalational. The infectious form, the spore, enters the body and is thought to germinate within macrophages either at the site of inoculation (cutaneous or orogastric) or in the regional lymph node (inhalational). The bacillus then synthesizes its antiphagocytic capsule and the lethal and oedema toxins which interfere with the non-specific host defences leading to the characteristic locally destructive lesion and spread by lymphatics to the systemic circulation and other organs. The cutaneous form begins as a papule which progresses over several days to a vesicle and then ulcerates. There is often oedema, sometimes massive, probably due to the oedema toxin that surrounds the lesions which then develop a characteristic black eschar. The patient may be febrile with mild to severe systemic symptoms of malaise, headache and toxicity. Oropharyngeal anthrax presents with severe sore throat or an ulcer in the oropharyngeal cavity associated with neck swelling, fever, toxicity and dysphagia. Gastrointestinal anthrax begins with anorexia, nausea, vomiting and abdominal pain which may be similar to an acute abdomen. There may be diarrhoea and ascites, both of which may be haemorrhagic. Inhalational anthrax begins with non-specific symptoms of malaise, fever, myalgia and non-productive cough. After a period of 2-3 days, this is followed by a sudden onset of severe respiratory distress associated with diaphoresis, cyanosis and increased chest pain. There may be a widened mediastinum and pleural effusions on chest X-ray. Death follows in 24-36 h from respiratory failure, sepsis and shock. The diagnosis of anthrax is easy if it is considered. The organism is readily observed by Gram or Wright stain in local lesions or blood smear and can be easily cultured from the blood and other body fluids. However, because of its rarity, it is not often included in the differential diagnosis and in inhalational disease the diagnosis is rarely made until the patient is moribund. More rapid diagnostic tests are under development. Penicillin, combined with supportive care, remains the mainstay of treatment, although the organism is susceptible in vitro to many antibiotics. In recent years, there have been significant advances in our knowledge of the organism and its toxins and it is anticipated that similar progress will be made in the future in developing more rapid diagnostic tests and new modalities of treatment. PMID- 10475976 TI - Immune correlates of protection against anthrax AB - Bacillus anthracis protective antigen (PA) has been produced from a recombinant B. subtilis and its efficacy, when combined with the Ribi adjuvant (MPL-TDW-CWS) or alhydrogel, has been compared with that of the licensed UK human vaccine, in guinea pigs challenged with aerosolized Ames strain spores. Recombinant PA combined with the Ribi adjuvant performed as well as PA from B. anthracis cultures in previous reports (Ivins & Welkos 1986; Ivins et al. 1990; Turnbull et al. 1991; Jones et al. 1996; McBride et al. 1998) giving protection in 100% of animals exposed to the highest challenge dose of the Ames strain of B. anthracis that can be administered practically (retained lung doses of approximately 106 spores). In attempts at identifying markers of protection in immunized individuals, rPA in combination with the Ribi adjuvant induced a marker IgG2 response in guinea pigs with no significant differences in IgG1 levels when compared with other vaccine formulations (McBride et al. 1998). In BALBc mice, rPA with the Ribi adjuvant induced a higher IgG2a response compared with rPA with anhydrogel and the human vaccine. To examine the role of anti-PA-specific antibodies in protection, guinea pig sera is being passively transferred into guinea pigs and SCID mice, followed by protection. Similarly, B- and T lymphocytes from immunized BALB/c mice are being separately and passively transferred into SCID mice with subsequent challenge. The neutralizing ability of the PA-specific antibodies is being studied using an in vitro macrophage lysis assay. PMID- 10475975 TI - In vitro correlate of immunity in an animal model of inhalational anthrax AB - The incidence of anthrax in humans is extremely low. Human vaccine efficacy studies for inhalational anthrax cannot be conducted. The identification of a correlate of protection that predicts vaccine efficacy is crucial for determining the immune status of immunized humans. This surrogate marker of immunity can only be established by using an appropriate animal model. Numerous studies showed that protective antigen (PA) is the principle protective antigen in naturally- or vaccine-induced immunity. However, attempts to correlate the quantity of anti-PA antibodies with protective immunity in the guinea pig model for anthrax and various vaccine formulations have failed. In these studies, we used the licensed anthrax vaccine adsorbed (AVA) in rabbits. Groups of New Zealand white rabbits, 10 or 20 per group, were immunized intramuscularly (two doses, 4 weeks apart) with varying doses of AVA, ranging from a human dose to 1:256 dilution in sterile phosphate-buffered saline (PBS). Control rabbits received PBS/Alhydrogel according to the same schedule. Each rabbit was bled 2 weeks after the second dose, and antibody levels to PA measured by both the quantitative anti-PA IgG ELISA and the toxin-neutralizing antibody (TNA) assay. Rabbits were aerosol challenged 10 weeks from day 0 with a lethal dose of Ames spores. All the rabbits that received the undiluted and 1:4 dilution of vaccine survived, whereas those receiving the higher dilutions of vaccine (1:16, 1:64 and 1:256) had deaths in their groups. All the controls died. Rabbit survival was compared with the antibody response. Statistical models were used to test for significance of the peak antibody responses to predict survival. Results showed that both the amount of anti-PA IgG and TNA titres present in the sera at the time of the peak antibody response were significant (P < 0.0001) predictors of survival. These results demonstrate that the humoral immune response to AVA can predict protection in the rabbit model of inhalational anthrax. PMID- 10475977 TI - Human immune responses to the UK human anthrax vaccine. AB - The IgG anti-protective antigen subclass antibody response of individuals who had been infected with anthrax was compared with that of healthy individuals immunized with the UK licensed anthrax vaccine. The predominant subclass in both groups was IgG1. In addition, IgG3 was seen in convalescent serum while vaccinees produced IgG2, IgG3 and IgG4 subclass. The significance of these results is discussed. Further work is required to determine the role of antibodies in mediating protective immunity in man. PMID- 10475978 TI - Expression of the protective antigen of Bacillus anthracis by Lactobacillus casei: towards the development of an oral vaccine against anthrax. AB - Bacillus anthracis is the causative organism of the disease anthrax. The ability of the organism to form resistant spores and infect via the aerosol route has led to it being considered as a potential biological warfare agent. The current available human vaccines are far from ideal, they are expensive to produce, require repeated doses and may invoke transient side-effects in some individuals. There is also evidence to suggest that they may not give full protection against all strains of B. anthracis. A new generation of anthrax vaccine is therefore needed. The use of Lactobacillus as a vector for expression of heterologous proteins from pathogens supplies us with a safe system, which can be given orally. Lactobacilli are commensals of the gut, generally regarded as safe and have intrinsic adjuvanticity. Oral vaccines may stimulate the mucosol immune system to produce local IgA responses in addition to systemic responses. These vectors are delivered at the mucosal surface, the site where the infection actually occurs and where the first line of defence lies. The gene encoding the protective antigen (PA) of B. anthracis, an immunogenic non-toxic component of the two toxins produced, is being cloned into different homologous vectors and subsequently transformed to various Lactobacillus strains. High intracellular expression levels for the PA in Lact. casei were achieved. Mucosal antigen presentation and humoral and cellular immune responses following immunization with transformants expressing PA in various ways (intracellular, surface-anchored and extracellular) are being studied. PMID- 10475979 TI - Presentation of protective antigen to the mouse immune system: immune sequelae. AB - Protective antigen (PA), the major protective component of the existing vaccine, is a potent immunogen. Protective antigen in alhydrogel induced a high serum IgG titre (> log10 4) in both the C57B16 and Balb/c mouse and the predominant subclass of antibody induced was IgG1, indicating that the response to PA was predominantly Th2 directed. When plasmid DNA encoding PA was used to immunize the Balb/c mouse, a low serum IgG titre was detected ( 0.999 for mercaptopurine and r > 0.998 for thioguanine) were satisfactory. The capsule excipients did not interfere with quantification of the mercaptopurine or thioguanine peak. CONCLUSION: The two antineoplastic agents could be assayed rapidly by the same capillary zone electrophoretic method with acceptable accuracy and precision. This method is suitable for routine control of capsules prepared for paediatric patients. PMID- 10475986 TI - Educational campaign to improve the prevention of postoperative venous thromboembolism. AB - BACKGROUND: Recent studies have revealed considerable scope for improvement in preventing postoperative venous thromboembolism. This project comprised a baseline assessment of the use of preventive measures within the Royal Hobart Hospital, followed by the implementation and evaluation of an educational program directed at hospital staff. AIM: To determine whether the Royal Hobart Hospital's guidelines for the prophylaxis of postoperative venous thromboembolism were being utilized effectively and, if necessary, attempt to improve use of the guidelines through an educational programme. METHODS: Data were collected retrospectively from the medical records of 250 surgical patients undergoing a procedure during February 1997. Patients were classified as being at a low, medium or high risk of venous thromboembolism using two sets of criteria. The percentage of patients receiving the appropriate prophylaxis was determined. An educational programme to promote the hospital's guidelines for the prophylaxis of postoperative venous thromboembolism was then implemented. Included were presentations to staff, posters placed throughout the hospital and the wide distribution of the results and a glossy card showing the hospital's guidelines. Follow-up data were subsequently collected from another 250 surgical patients. RESULTS: Only 59% of patients received appropriate prophylaxis according to the hospital's approved guidelines, with little change when those patients with possible contraindications to thromboprophylaxis were excluded. When those patients at a high risk of venous thromboembolism were examined, only 25% were prescribed the recommended preventive measures. There was no difference between elective versus emergency surgery and the use of appropriate prophylaxis. Following the implementation of the educational programme, data collection from another 250 surgical patients revealed a significant increase (P < 0.05 by chi-square test) in the level of appropriate thromboprophylaxis to 70% of patients, with 77% of the high-risk patients being prescribed the recommended preventive measures. CONCLUSIONS: A pharmacy-based educational intervention significantly improved adherence to the hospital's guidelines for the prophylaxis of postoperative venous thromboembolism. PMID- 10475987 TI - Programme to improve the use of drugs in older people and involve general practitioners in community education. AB - As part of a multi-pronged approach to improving the quality of drug use in the elderly, a pharmacist was contracted by the Division of General Practice (Northern Tasmania) to develop educational material and implement two academic detailing sessions for general practitioners on the issues of adverse drug reactions and drug interactions in older people. The project aimed to involve general practitioners in community education after optimizing relevant therapeutic knowledge and standardizing prescribing practices. Sixteen general practitioners were involved in the project and 13 of these agreed to participate in academic detailing. The pharmacist developed prescribing guidelines for general practitioners and discussed these and illustrative case studies at the academic detailing sessions. General practitioner-conducted education sessions were completed by nine general practitioners to groups of carers, general practitioners, nurses and older people. Despite the relatively low numbers of general practitioners involved, the results of the project were encouraging. Academic detailing by the pharmacist was well received by the general practitioners, who indicated they would be willing to participate in further sessions. Pre- and post-project multiple-choice tests on therapeutic issues in the elderly indicated a strong trend for an increase in knowledge. Analysis of general practitioners' patient records found a statistically significant decline in the median number of medications prescribed per patient during the project. There was also a statistically significant decline in prescribing of 'indicator' medications, particularly psychoactive drugs and nonsteroidal anti-inflammatory drugs in patients resident in nursing homes. The project demonstrated that academic detailing by a pharmacist can be effective as part of a combined approach to improve the quality of drug use in older people. PMID- 10475988 TI - Stability of ceftazidime in a viscous eye drop formulation. AB - OBJECTIVE: To assess the stability of an extemporaneously prepared ceftazidime eye-drop. METHOD: Ceftazidime was formulated at a concentration of 5% w/v as an eye drop, using Sno Tears, an artificial tear solution containing polyvinyl alcohol, as a vehicle. Two batches of the formulation were stored in 10 ml eye drop bottles at 7 and 25 degrees C for up to 14 days. Ceftazidime and pyridine, its degradation product, were determined at intervals by HPLC. RESULTS: A yellow coloration was evident after 7 days at 7 degrees C and after 24 h at 25 degrees C. Ceftazidime lost approximately 35% after 7 days storage at 25 degrees C. At 7 degrees C, the mean time to 10% degradation, determined by linear regression, was 11 and 8 days for the two batches. However, the lower 95% confidence limits were 8 and 5 days, respectively. Pyridine levels increased during storage. The mass balance between ceftazidime remaining and pyridine formed was close to 100% during the early part of storage. By the end of storage, the balance had reduced to around 95% at 7 degrees C and 80% at 25 degrees C. This discrepancy may be due to sorption of pyridine to the butyl rubber bottle closure. The pH remained in the range 6-7 throughout the storage period. CONCLUSION: The formulation may be stored for 5 days in the refrigerator. PMID- 10475989 TI - An evaluation of prostaglandin E2 vaginal gel use in practice. AB - OBJECTIVE: The purpose of this study was to investigate the effectiveness of prostaglandin E2 vaginal gel as used in practice, rather than its efficacy as assessed in randomised, controlled, clinical trials. This product is used to ripen the cervix prior to induction of labour, sometimes making unnecessary the use of the standard treatment for induction, artificial rupture of the membranes (ARM) plus oxytocin. In this study, effectiveness of the gel was assessed in terms of changes in mode of delivery, and in particular the risk of Caesarean section. METHODS: An historical control was used and the risk of Caesarean section for women induced in the 1990/91 (before the introduction of the gel) was compared with that for women induced in 1992/93 (after the introduction of the gel). Maternal characteristics which may have been different in the two groups and factors which might influence the risk of Caesarean section were controlled for statistically using logistic regression, thus reducing any bias towards one group. RESULTS: After adjusting for the factors which had a significant effect on the process of labour from induction to birth, it was found that the risk of Caesarean section was not significantly lower in the 1992/93-time period, when the gel was in regular use, from that in the 2 years prior to its introduction (Odds ratio 1.09, CI95% 0.88, 1.36). CONCLUSION: Following the introduction of PGE2 gel, no difference in effectiveness, as measured in terms of mode of delivery, was detected in this study of practice, which included patients with more complex obstetric problems. PMID- 10475990 TI - Turn of tide for oral vitamin B12 treatment. PMID- 10475991 TI - Bleeding complications in secondary stroke prevention by antiplatelet therapy: a benefit-risk analysis. AB - This review analyses the benefit-risk ratio of antiplatelet drugs in secondary stroke prevention and is based on the published data from eight large stroke prevention trials. In patients with prior transient ischaemic attack (TIA) or stroke, aspirin prevented one to two vascular events (stroke, AMI, or vascular death) per 100 treatment-years with an excess risk of fatal and severe bleeds of 0. 4-0.6 per 100 treatment-years. The gastrointestinal bleeding risk was significantly lower with ticlopidine and clopidogrel, which were both somewhat more effective than aspirin in the prevention of vascular events. The combination of dipyridamole and aspirin prevented 2.82 strokes at the expense of an excess risk of 0.61 (95% CI = 0.27-0.95) fatal or severe bleeds per 100 treatment-years. In the acute phase of stroke, the aspirin-associated risk of haemorrhagic complications was much increased compared with that in the stable phase after stroke, with 0.48 (95% CI = 0.13-0.83) fatal or severe bleeds per 100 treated patients for the first 4 weeks after stroke in the Chinese Acute Stroke Trial and 0.41 (95% CI = 0. 05-0.77) in the International Stroke Trial. Still, there was a net benefit with the prevention of about one death or non-fatal ischaemic stroke per 100 treated patients. PMID- 10475992 TI - A rise in plasma creatinine that is not a sign of renal failure: which drugs can be responsible? AB - This is a review of the available information about drugs which cause an increase in plasma creatinine concentration without decreasing glomerular filtration rate (GFR). The GFR is the main, but not the single, determinant of the plasma creatinine levels. Several drugs, such as cimetidine, trimethoprim, corticosteroids, pyrimethamine, phenacemide, salicylates and active vitamin D metabolites, have been reported to increase plasma creatinine without influencing its glomerular filtration. Cimetidine, trimethoprim, pyrimethamine and salicylates can inhibit secretion of creatinine by the proximal tubule. Corticosteroids and vitamin D metabolites probably modify the production rate and the release of creatinine. The exact mechanism of phenacemide-creatinine interaction is not fully explained. These drug-induced alterations in plasma creatinine concentration have clinical significance when GFR is estimated by using plasma creatinine. PMID- 10475993 TI - Intensive rehabilitation of emotionally distressed patients after coronary by pass grafting. AB - OBJECTIVES: To evaluate whether the status of emotionally distressed post coronary by-pass surgery patients can be improved by a comprehensive, in-patient rehabilitation programme. DESIGN: Cross-sectional. SETTING: Rehabilitation hospital. SUBJECTS: One hundred and fifty-two post coronary by-pass surgery patients referred to an intensive rehabilitation programme. The study group was divided into two subgroups, according to the level of emotional distress. Forty three (30%) out of 142 patients who completed the Nottingham Health Profile were considered to be distressed. INTERVENTIONS: The comprehensive 4-week inpatient rehabilitation programme consisted of daily physical exercise, lectures about coronary disease and risk factors, psychological support and nutrition counselling. MAIN OUTCOME MEASURES: Physical fitness, blood lipids and quality of life questionnaires. RESULTS: The two-way analysis of variance showed that emotionally distressed patients achieved equally good results as those obtained by nondistressed patients in performance regarding the maximal exercise stress test, maximum rate pressure product achieved, or the level of resting heart rate. Significant improvement in psychological well-being as assessed by a seven-point rating scale was observed in both groups. Both groups of patients were also equally successful in weight reduction and lowering of total cholesterol and triglycerides. Of all the patients who were employed at admission, twice the number of patients in the distressed group were in receipt of a permanent disability pension and half the number of patients was employed, compared to the initially nondistressed group, at 1-year follow-up. CONCLUSIONS: Distressed patients were very successful in improving their functional status and reducing risk factors when offered an intensive rehabilitation programme but they were much less successful in returning to work. Secondly, patients who were emotionally distressed after surgery did not differ in regard to disease status or physical capacity from nondistressed patients. They did, however, experience more angina pectoris both in daily life and when exposed to a maximal exercise stress test. Finally, the presence of emotional distress was easily detected by any measure of psychological status. PMID- 10475994 TI - Effects of age, gender and metabolic factors on endothelium-dependent vasodilation: a population-based study. AB - OBJECTIVES: A progressive decline in endothelium-dependent vasodilation (EDV) in the human forearm with age has previously been reported. The aim of this study was to evaluate the interplay between age, gender and metabolic factors on EDV in healthy subjects in a population-based study. SETTING: Tertiary university hospital. SUBJECTS AND DESIGN: Thirty-six healthy men and 30 women, aged 20-69 years, underwent measurements of forearm blood flow (FBF) at rest and during local infusions of 2 and 4 microg min-1 of metacholine (evaluating EDV) and 5 and 10 microg min-1 of sodium nitroprusside (evaluating endothelium-independent vasodilation, EIDV) and during reactive hyperaemia by venous occlusion plethysmography. RESULTS: Age was inversely related to EDV (r = - 0.41, P < 0.05 in men; r = - 0.61, P < 0.01 in women) and maximal FBF during reactive hyperaemia in both men and women. EIDV was significantly related to age in an inverse way in women only. EDV was more pronounced in females than in males before menopause (48 +/- 3 SD years, 635 +/- 186 vs. 502 +/- 269% in males, P < 0.05), but similar in women and men thereafter (374 +/- 141 vs. 370 +/- 185% in men). The slope of the regression line for the relationship between age and EDV was flatter in premenopausal than in postmenopausal women (- 2.3 vs. - 6.4), whilst this slope was similar in younger and older men (- 5.5 vs. - 5.3). In multiple regression analysis, fasting blood glucose levels and the waist/hip ratio remained the only significant predictors of EDV in men (P < 0.01 for both), whilst age was the only significant independent predictor of EDV in women (P < 0.01). CONCLUSION: The interplay between age and metabolic factors as determinants of endothelial function is different in healthy men and women. PMID- 10475995 TI - The health care costs of heart failure in Sweden. AB - AIM: Heart failure is a common and serious condition requiring extensive health care resources. The aim of this study is to estimate the total treatment costs of heart failure in Sweden. METHODS AND RESULTS: The study is a prevalence-based cost-of-illness study. It includes costs of institutional care (hospitals and nursing homes), outpatient care, surgery and drugs. The costs are estimated based on official Swedish statistics, and on various clinical and epidemiological studies. The results are expressed in 1996 prices. The total annual treatment costs for heart failure are approximately Swedish kronor (SEK) 2000-2600 million, or nearly 2% of the Swedish health care budget. Institutional care is the single largest component, amounting to SEK 1300-1900 million, or about 65-75% of the costs of heart failure treatment. CONCLUSIONS: The results from this study indicate that heart failure is a costly condition. Efforts to develop effective management programmes that can reduce the need for expensive institutional care, without a negative impact on quality of life, morbidity and mortality, should be given high priority. PMID- 10475996 TI - Declining prevalence of angina pectoris in middle-aged men and women. A population-based study within the Northern Sweden MONICA Project. Multinational Monitoring of Trends and Cardiovascular Disease. AB - OBJECTIVES: To describe trends in the prevalence of angina pectoris in northern Sweden, between 1986 and 1994. DESIGN: Cross-sectional population studies. SETTING: Northern Sweden MONICA Project in Norrbotten and Vasterbotten counties, Sweden. SUBJECTS: Randomly selected men and women in the age group 35-64 years, total of 2459 men and women. MAIN OUTCOME MEASURES: Comparison of the prevalence of angina pectoris in 1986 and 1994 as measured by the Rose questionnaire. RESULTS: The proportion with a history of myocardial infarction decreased amongst the participants from 4.6% to 2.0% (P < 0.001) between 1986 and 1994. The prevalence of angina pectoris in men was essentially unchanged (3.4% in 1986 to 3.1% in 1994 (chi2 = 0.02; P = 0.87), whereas it declined significantly in women from 5. 9% to 2.8% (chi2 = 6.32; P = 0.01). In both men and women, the highest prevalence of Rose-positive persons was found in the oldest age group. In 1986 the Rose-positive subgroup had a significantly higher proportion with high cholesterol (>/=6.5 mmol L-1) as compared with the Rose-negative subgroup, 64% vs. 48% (chi2 = 5.04; P = 0.02). In both surveys high blood pressure was more common in the Rose-positive group (1986: chi2 = 13.2; P < 0.001 and 1994: chi2 = 9.8; P = 0.002). CONCLUSIONS: In women, but not in men, the prevalence of angina pectoris decreased significantly between 1986 and 1994. During the same time period the proportion of people with high cholesterol decreased in northern Sweden. In both surveys, individuals with angina pectoris had more frequent hypertension. PMID- 10475997 TI - The measurement of venous haematocrit in patients with polycythaemia vera. AB - OBJECTIVE: In clinical practice, patients with polycythaemia vera (PV) are monitored by measurement of venous packed cell volume (PCV). However, whereas treatment recommendations are still based upon studies in which the results were obtained with the centrifuged microhaematocrit, currently in most instances automated blood cell counters are used to calculate PCV. In a group of patients with polycythaemia we therefore compared the results obtained by the microhaematocrit method with PCV calculated by haematology analysers. DESIGN: The study was carried out on a prospective basis. Duplicate venous blood samples were collected. The centrifuged microhaemotocrit was obtained by using an IEC Micro-MB Centrifuge. Depending on different routine methods used in the participating hospitals, the blood cell counter PCV was calculated using Coulter STKS, Bayer Technicon H2 or H3. SETTING: Patients were included from four Swedish university hospitals: Akademiska (Uppsala), Huddinge and Karolinska (Stockholm) and Sahlgrenska (Goteborg). SUBJECTS: Seventy-four patients with PV and 10 patients with secondary polycythaemia were included and a total of 150 duplicate blood samples were analysed from these subjects. RESULTS: In the 150 measurements the mean blood cell counter calculated PCV was 0.448 +/- 0.037; the mean for centrifuged microhaematocrit was 0.467 +/- 0. 037 and the difference between means was highly significant (P = 6.8 x 10-25). The means for centrifuged haematocrit and calculated PCV differed significantly in the groups of PV patients treated with phlebotomy only, hydroxyurea or radiophosphorous (P < 0.0001, respectively). In PV patients treated with alpha-interferon and in patients with secondary polycythaemia the difference in means did not reach statistical significance (P = 0.07 and P = 0.13, respectively). The groups of patients with MCV <80 fL and >/=80 fL both presented significant differences between means for calculated PCV and centrifuged haematocrit. CONCLUSIONS: If PV patients are monitored with blood cell counter calculated PCV it appears that the therapeutic goal should be to maintain the calculated PCV below 0.43, provided the local differences in calculated PCV and centrifuged haematocrit are of the same magnitude as in this study. PMID- 10475998 TI - Increased QT dispersion during hypoglycaemia in patients with type 2 diabetes mellitus. AB - OBJECTIVES: To study effects of insulin-induced hypoglycaemia on the cardiac repolarization, using QT interval measurements, in patients with type 2 diabetes. DESIGN: Hypoglycaemia was induced by an i.v. insulin-infusion and blood glucose was clamped at 2.7 mmol L-1 for 60 min (T = 90-150 min) in two experiments, with (+GLIB) and without (-GLIB) glibenclamide. In a third experiment, with similar hyperinsulinaemia, glucose was clamped at a euglycaemic level (;5 mmol L-1). ECG was continuously recorded for arrhythmia-monitoring, and 12-lead ECGs were recorded at T = 0 and 150 min. QT intervals were measured, and we determined QT dispersion (difference between the maximum and the minimum QT interval) reflecting interlead variability of repolarization. SUBJECTS: Thirteen patients with type 2 diabetes, on combined insulin and glibenclamide treatment, were studied during hypoglycaemia, and eight of them participated in the euglycaemic experiment. RESULTS: No significant arrhythmias were seen during hypoglycaemia but the mean QT intervals and QT dispersion increased significantly (P < 0.001), with no differences between -GLIB and +GLIB. During the euglycaemic clamp all QT measurements remained unchanged. Serum potassium decreased significantly (P < 0.001) during all three clamps, but the decrease was more pronounced during hypoglycaemia. The change in potassium was not correlated to the degree of QT prolongation or QT dispersion. CONCLUSIONS: Significant changes in the repolarization of the heart can be seen during hypoglycaemia in patients with type 2 diabetes, indicating an increased risk of arrhythmia at low blood glucose levels. PMID- 10475999 TI - Quality of life after cerebrovascular stroke: a systematic study of patients' preferences for different functional outcomes. AB - OBJECTIVES: To elicit valid quality of life estimates and the highest acceptable treatment risk of different outcomes after stroke. This is a prerequisite for rational medical decision-making, especially when considering treatments like thrombolysis. SUBJECTS: Healthy people, non-stroke medical patients and stroke survivors aged 20-84 years (n = 158) INTERVENTIONS: Subjects were interviewed by a physician using three different methods ('standard gamble', 'time trade-off' and 'direct scaling') supported by an interactive computer program. MAIN OUTCOME MEASURES: We measured utility, a numerical value ranging from 0.00 (death) to 1.00 (perfect health), representing the strength of the patient's preference for an outcome. When using the standard gamble method, risk is also introduced into the measurement. RESULTS: People's preferences for stroke outcomes varied widely, and the estimates were influenced by assessment method. We found that previous stroke, marital status and age were the only independent variables influencing the utility given. Subjects in our population over the age of 45 were very comparable to the real population at risk for acute stroke regarding these three variables, and they assigned a median utility of 0.91 (10th percentile, 0.65; 90th percentile, 0.99) to a minor stroke and 0.61 (10th percentile, 0.08; 90th percentile, 0.95) to a major stroke using the standard gamble method. CONCLUSIONS: Most people do not feel that suffering from stroke is an overwhelming catastrophe and they do not accept treatment options with very high risks. PMID- 10476000 TI - Elevated serum levels of methylmalonic acid and homocysteine in elderly people. A population-based intervention study. AB - OBJECTIVES: There is uncertainty amongst clinicians about the definitions of cobalamin and folate deficiency and therefore about the indications for treatment. In this report we present the results of systematic cobalamin and folic acid treatment based upon serum cobalamin, total homocysteine (tHcy) and methylmalonic acid (MMA) analyses in a population-based sample. SUBJECTS: A 20% random sample of persons 70 years or older in a defined geographical area were invited to a survey (n = 266). Sixty-nine persons who had serum cobalamin < 300 pmol L-1 and serum MMA >/=0.37 micromol L-1 or serum tHcy >/=15 micromol L-1 and who had no cobalamin or folic acid substitution were selected for treatment. INTERVENTIONS: Initially all 69 patients were given cobalamin orally or intramuscularly. Those who remained high in tHcy were in addition given folic acid treatment. MAIN OUTCOME MEASURES: Serum cobalamin, serum MMA and serum tHcy. RESULTS: After 6 months of cobalamin treatment, serum MMA became normal in 13 out of 15 persons. Mean serum tHcy decreased but was normalized in only 15 out of 56 persons. After 3 months of folic acid treatment added to those who still had an abnormal serum tHcy, serum tHcy had normalized in all but one person. CONCLUSIONS: Cobalamin treatment normalizes increased MMA values and combined cobalamin and folic acid treatment normalizes tHcy, suggesting a pretreatment deficiency of tissue cobalamin and folate in spite of normal serum cobalamin and folate values in the majority of cases. PMID- 10476001 TI - Heparin-induced thrombocytopenia (HIT) due to heparin flushes: a report of three cases. AB - Three cases of heparin-induced thrombocytopenia (HIT) are reported that were provoked by daily heparin flushes of central venous access devices. Each case had confounding features that delayed recognition of the problem. A review of the literature revealed only 29 previously reported cases of HIT secondary to heparin flushes. However, the true incidence of this problem is unknown. A high index of suspicion and confirmatory laboratory tests are necessary to make the diagnosis. PMID- 10476002 TI - Effects of light exposure and sleep displacement on dim light melatonin onset. AB - The purpose of the study was to induce in two different ways, a phase-angle difference between the circadian pacemaker and the imposed sleep-wake cycle in humans, we intended to: (i) shift the circadian pacemaker by exposure to bright light and keep the timing of the sleep-wake cycle fixed; and (ii) keep the timing of the circadian pacemaker fixed by a constant light-dark cycle and displace sleep. We monitored dim light melatonin onset (DLMO), core body temperature and sleep. DLMO was delayed significantly after 3 days of a 3-h delayed sleep-phase when compared with 3 days of sleep at a normal or 3-h advanced sleep-phase. The shifts in DLMO were not accompanied by shifts in body temperature, changes in waking-up time or by a change in the duration of the first rapid eye movement (REM) sleep episode. Three days of light exposure in the morning or evening resulted in shifts in DLMO of similar magnitude, but this was accompanied by shifts in the rhythm of body temperature, changes in waking-up time and in the duration of the first REM sleep episode. We conclude that the changes observed after light exposure reflect shifts in the circadian pacemaker. In contrast, we propose that the changes observed in DLMO after sleep displacement are not mediated by the circadian pacemaker. These results raise some doubts about the reliability of DLMO as a marker of circadian phase in cases of sleep disturbances. Finally, we initiate a search for changes in sleep that might be responsible for the unexpected effects on DLMO. PMID- 10476003 TI - Comparison between subjective and actigraphic measurement of sleep and sleep rhythms. AB - Sleep is often assessed in circadian rhythm studies and long-term monitoring is required to detect any changes in sleep over time. The present study aims to investigate the ability of the two most commonly employed methods, actigraphy and sleep logs, to identify circadian sleep/wake disorders and measure changes in sleep patterns over time. In addition, the study assesses whether sleep measured by both methods shows the same relationship with an established circadian phase marker, urinary 6-sulphatoxymelatonin. A total of 49 registered blind subjects with different types of circadian rhythms were studied daily for at least four weeks. Grouped analysis of all study days for all subjects was performed for all sleep parameters (1062-1150 days data per sleep parameter). Good correlations were observed when comparing the measurement of sleep timing and duration (sleep onset, sleep offset, night sleep duration, day-time nap duration). However, the methods were poorly correlated in their assessment of transitions between sleep and wake states (sleep latency, number and duration of night awakenings, number of day-time naps). There were also large and inconsistent differences in the measurement of the absolute sleep parameters. Overall, actigraphs recorded a shorter sleep latency, advanced onset time, increased number and duration of night awakenings, delayed offset, increased night sleep duration and increased number and duration of naps compared with the subjective sleep logs. Despite this, there was good agreement between the methods for measuring changes in sleep patterns over time. In particular, the methods agreed when assessing changes in sleep in relation to a circadian phase marker (the 6-sulphatoxymelatonin (aMT6s) rhythm) in both entrained (n = 30) and free-running (n = 4) subjects. PMID- 10476004 TI - Word recall correlates with sleep cycles in elderly subjects. AB - Morning recall of words presented before sleep was studied in relation to intervening night sleep measures in elderly subjects. Night sleep of 30 elderly subjects aged 61-75 years was recorded. Before sleep, subjects were presented with a list of paired non-related words and cued recall was asked immediately after the morning awakening. Recall positively correlated with average duration of NREM/REM cycles, and with the proportion of time spent in cycles (TCT) over total sleep time (TST). No significant correlations were found with other sleep or wake measures. These results suggest the importance of sleep structure for sleep-related memory processes in elderly adults. PMID- 10476005 TI - Circadian variation of EEG power spectra in NREM and REM sleep in humans: dissociation from body temperature. AB - In humans, EEG power spectra in REM and NREM sleep, as well as characteristics of sleep spindles such as their duration, amplitude, frequency and incidence, vary with circadian phase. Recently it has been hypothesized that circadian variations in EEG spectra in humans are caused by variations in brain or body temperature and may not represent phenomena relevant to sleep regulatory processes. To test this directly, a further analysis of EEG power spectra - collected in a forced desynchrony protocol in which sleep episodes were scheduled to a 28-h period while the rhythms of body temperature and plasma melatonin were oscillating at their near 24-h period - was carried out. EEG power spectra were computed for NREM and REM sleep occurring between 90-120 and 270-300 degrees of the circadian melatonin rhythm, i.e. just after the clearance of melatonin from plasma in the 'morning' and just after the 'evening' increase in melatonin secretion. Average body temperatures during scheduled sleep at these two circadian phases were identical (36.72 degrees C). Despite identical body temperatures, the power spectra in NREM sleep were very different at these two circadian phases. EEG activity in the low frequency spindle range was significantly and markedly enhanced after the evening increase in plasma melatonin as compared to the morning phase. For REM sleep, significant differences in power spectra during these two circadian phases, in particular in the alpha range, were also observed. The results confirm that EEG power spectra in NREM and REM sleep vary with circadian phase, suggesting that the direct contribution of temperature to the circadian variation in EEG power spectra is absent or only minor, and are at variance with the hypothesis that circadian variations in EEG power spectra are caused by variations in temperature. PMID- 10476006 TI - The effects of leptin on REM sleep and slow wave delta in rats are reversed by food deprivation. AB - Leptin (ob protein) is an adipose tissue derived circulating hormone that acts at specific receptors in the hypothalamus to reduce food intake. The protein is also critically involved in energy balance and metabolic status. Here the effect of leptin on sleep architecture in rats was evaluated because food consumption and metabolic status are known to influence sleep. Sprague-Dawley rats were chronically implanted with electrodes for EEG and EMG recording and diurnal sleep parameters were quantified over 9-h periods following leptin administration. Murine recombinant leptin (rMuLep) was administered systemically to rats that either had undergone 18 h of prior food deprivation or had received food ad libitum. In the normally fed rats, leptin significantly decreased the duration of rapid eye movement sleep (REMS) by about 30% and increased the duration of slow wave sleep (SWS) by about 13%, the latter effect reflecting enhanced power in the delta frequency band. These results are consistent with studies that have linked changes in metabolic rate with effects on sleep. Leptin administration has previously been shown to alter neuroendocrine parameters that could have mediated these changes in sleep architecture. Unexpectedly, prior food deprivation negated the effect of leptin on both REMS and SWS, a result that emphasizes the significance of the apparent coupling between sleep parameters and energy status. PMID- 10476007 TI - Atrial natriuretic peptide levels and pulmonary artery pressure awake, at exercise and asleep in obstructive sleep apnoea syndrome. AB - Elevated nocturnal plasma atrial natriuretic peptide (ANP) levels were found in patients with obstructive sleep apnoea (OSA). The purpose of our study was to examine the secretion of ANP during the night and to measure changes in oxygen saturation, pulmonary artery pressure and intrathoracic pressure swings in patients with OSA. Moreover, we analysed the secretion of ANP and the pulmonary artery pressure in different behavioural states, e.g. awake, at exercise and asleep. Consecutive apnoeas in non-rapid eye movement (NREM) sleep at the beginning, middle and end of the sleep study were analysed in six patients with obstructive sleep apnoea. In addition, we measured the plasma levels of ANP. The apnoea duration was significantly longer (P< 0.05) at the middle of the sleep study than at the beginning or end. Correspondingly, the end-apnoeic oxygen saturation and end-apnoeic oesophageal pressure were both significantly lower (P< 0.05) in the middle of the sleep study than at the beginning or end. No significant differences were found in the end-apnoeic systolic transmural pulmonary artery pressure (P(PATM)) and the levels of ANP. Evaluation of the ANP levels during different behavioural states revealed that the asleep levels were slightly, but not significantly, higher than the awake levels (0.235+/-0.088 vs. 0.207+/-0.057 nmol/L). However, the highest levels were found during exercise (0.334+/-0.170 nmol/L) with a significant difference compared with the awake and asleep levels. These data suggest that volume effects may be a potent factor in liberating ANP during exercise, but the role of OSA in ANP secretion when asleep is questionable. PMID- 10476008 TI - Periodic limb movements and obstructive sleep apneas before and after continuous positive airway pressure treatment. AB - Periodic limb movements during sleep (PLMS) and obstructive sleep apnea syndrome (OSAS) are two common sleep disorders. The similarity in periodicity of periodic limb movements (PLMs) and obstructive sleep apneas (OSAs) led us to hypothesize the existence of a common central generator responsible for the periodicity of both OSAs and PLMs. In order to test this hypothesis, we compared apnea periodicity before continuous positive airway pressure (CPAP) treatment with PLMs periodicity during CPAP treatment in 26 OSA patients, consecutively recorded and treated in our sleep laboratory. The investigation on CPAP was performed twice, once during the initial evaluation and once during a follow-up evaluation after 3 months of home treatment with CPAP. Our results showed that, in this sample, 16 patients out of 26 had an association of OSAS and PLMS, defined as the occurrence of at least 5 PLMs per hour of sleep. The mean apnea interval - measured as the time between the beginning of two successive apneas - was 43.1 s (+/-15.2, SD) and the mean PLM interval - calculated in the same way - was 29.6 s (+/-15.2) during the baseline night, 28.5 s (+/-15.7) during the first CPAP night, and 29.8 s (+/-14.8) during the second CPAP night. Thus, the periodicity of the two phenomena (apneas and PLMs) was different, both before and after CPAP treatment (P< 0.05). When considering the interval between the end of an event (apnea or PLM) and the beginning of the next one the mean apnea interval was 19.5 s (+/-11. 6), and the mean PLM interval was 28.1 s (+/-15.3) during the untreated night, 26.6 s (+/-16) during the first CPAP night and 27.9 s (+/-15) during the second CPAP night. The shortening of apnea intervals with this method of measuring intervals reflects the longer duration of apneas as compared to PLMs. Again the intervals between PLMs were not different between each other but the intervals between apneas were different from the intervals between PLMs (P< 0. 05) These results show that the periodicity of PLMs is different from that of OSAs, suggesting that sleep apneas and PLMs are not generated by a common central generator. PMID- 10476009 TI - Sleep/wake abnormalities in patients with periodic leg movements during sleep: factor analysis on data from 24-h ambulatory polygraphy. AB - Periodic Leg Movements (PLM) in sleep occur in a wide variety of sleep/wake disorders but their relationship with sleep disturbance, and notably with the concomitant existence of a 'restless legs' syndrome (RLS) remains unclear. We performed 24-h ambulatory polygraphy in a population of 54 consecutive, unselected patients with PLMs (Coleman's index greater than 5/h) who complained of different sleep disorders. A Principal Component Analysis (PCA) was conducted on seven variables from the sample, namely PLM index, patient's age, sleep stage changes per hour, sleep depth index (SWS+PS/TST), diurnal sleep time, number of awakenings exceeding 2 min and presence of a RLS. PCA yielded four independent factors. The PLM index and the changes of sleep stage clustered in a single factor, linking therefore sleep fragmentation to the frequency of PLMs. The second factor appeared to reflect a circadian sleep/wake disorder, combining diurnal sleep time with the number of long night awakenings. The third factor was mainly loaded by the patients' age and the sleep depth index, thus reflecting a well known relationship. Finally, the variable reflecting the existence of a RLS appeared isolated in a single factor, independent from the three previously described. These results confirm and extend the link between PLMs and sleep fragmentation, as well as the recently described dissociation between PLMs and diurnal somnolence. On the other hand, our analysis suggests that in PLM patients the concomitant existence of a RLS is not related to the frequency of occurrence of PLMs, at least when these latter are quantified independently of their arousal index. PMID- 10476011 TI - Multiprofessional working: learning from experience. PMID- 10476010 TI - Reversal of the sleep/wake cycle disorder of sleeping sickness after trypanosomicide treatment. AB - To determine whether the circadian disruption of the sleep/wake cycle observed in sleeping sickness, human African trypanosomiasis (HAT), can be reversed after trypanosomicide treatment, 10 Congolese patients infected by Trypanosoma brucei gambiense underwent 24-h polysomnographic recordings before treatment with melarsoprol and after each of three weekly treatment sessions. Polysomnography consisted of a continuous recording of the electroencephalogram, electromyogram and electro-oculogram on a Minidix Alvar polygraph. Sleep traces were analysed in 20-sec epochs for wakefulness, REM sleep, and NREM sleep [stages 1, 2, 3, 4; stages 3 and 4 representing slow-wave sleep (SWS)]. As previously described (Buguet et al. 1993), the 24-h distribution of the sleep/wake cycle was disturbed proportionally to the severity of the illness. The overall amounts of each sleep/wake stage did not change after treatment. However, the patterns of occurrence of sleep episodes, REM sleep and SWS phases were determinant in the evaluation of treatment efficacy. The trypanosomicide action of melarsoprol led to a reduction in the number of sleep episodes, except in one patient whose health condition worsened during the third treatment session: sleep onset REM sleep phases (SOREMPs) decreased and the number of SWS episodes during a sleep episode increased. We conclude that in HAT, the reversibility of the sleep/wake cycle alteration and that of sleep structure constitute the basis for an evaluation of the healing process. PMID- 10476012 TI - Patients in medical education: from passive participants to active partners. PMID- 10476013 TI - Teacher development in hospital medicine and dentistry. PMID- 10476014 TI - Objectives for the undergraduate teaching of psychiatry: survey of doctors and students. AB - OBJECTIVES: The principal aim was to assess the psychiatric topics that doctors and students considered most important for undergraduate teaching. Differences between doctors and students, men and women, physicians/students with or without an interest in psychiatry were examined. DESIGN: A mailed questionnaire was used concerning the knowledge and skills of psychological/psychiatric medi- cine considered to be needed in medical practice. SETTING: The Medical School of the University of Geneva. SUBJECTS: Doctors and undergraduate medical students in their last 2 years of medical training. RESULTS: Both doctors and students agreed on most topics, even though the students tended to give all items a higher rating. Both groups agreed on the importance of the following main topics: the doctor-patient relationship, identification and management of the principal psychiatric disorders and their associated risks and problems of a psychosocial nature. Those doctors showing an interest in psychiatry tended to accentuate the importance attached to interpersonal skills. The male and female doctors and students expressed very similar opinions. The female doctors, however, tended to attach greater importance to relational-emotional aspects and to disorders affecting children and adolescents than did their male colleagues, which is probably a reflection of the specific role that women still play within our society. When asked to assess the current teaching they received in medical school, the students considered that certain important aspects of psychiatry were insufficiently taught. CONCLUSION: These results confirm the importance of teaching psychiatry with an emphasis on problems encountered in general practice. PMID- 10476015 TI - Do morning report format changes affect educational content? AB - OBJECTIVE: The goal of this prospective observational study was to determine if format changes in a paediatric morning report conference affected the educational content presented. DESIGN: Case presentations during paediatric morning report at the University of Iowa during the academic years 1995-96 (Format 1 - informal presentations) and 1996-97 (Format 2 - formal presentations) were analysed for demographics, clinical venues where patients were encountered, case diagnoses and ensuing discussion. A 10-item questionnaire of participants' attitudes was conducted during May 1997. SETTING: Department of Pediatrics, University of Iowa. SUBJECTS: Paediatric residency trainees. RESULTS: The number of cases was significantly decreased from 4 to 3.2 per morning report session. A wide variety of patient ages and all clinical venues were represented in both formats. Case diagnoses covered 30 of 31 Pediatrics Review and Education Program (PREP, American Academy of Pediatrics, Elk Grove Village, IL, USA) Content Specification Headings during Format 1 vs. 29 of 31 during Format 2. The most common headings were the same. Patient evaluations were discussed less and patient clinical presentations and pathophysiology were discussed more during Format 2. Participants rated Format 2 higher; 54% of participants wanted to continue this format and 16% wanted a combination of both formats. CONCLUSIONS: This research suggests that while format changes in paediatric morning report resulted in a 20% decrease in the number of cases presented, only relatively small changes in the overall educational content occurred. Changes in educational programmes should be accompanied by educational evaluation. PMID- 10476016 TI - Teaching clinically experienced physicians communication skills. A review of evaluation studies. AB - CONTEXT: Interest in the teaching of communication skills in medical schools has increased since the early seventies but, despite this growing interest, relatively limited curricular time is spent on the teaching of communication skills. The limited attention to the teaching of these skills applies even more to the physicians' clinical years, when attention becomes highly focused on biomedical and technical competence. Continuing training after medical school is necessary to refresh knowledge and skills, to prohibit decline of performance and to establish further improvements. OBJECTIVE: This review provides an overview of evaluation studies of communication skills training programmes for clinically experienced physicians who have finished their undergraduate medical education. The review focuses on the training objectives, the applied educational methods, the evaluation methodology and instruments, and training results. METHODS: CD-ROM searches were performed on MedLine and Psychlit, with a focus on effect-studies dating from 1985. RESULTS: Fifteen papers on 14 evaluation studies were located. There appears to be some consistency in the aims and methods of the training programmes. Course effect measurements include physician self-ratings, independent behavioural observations and patient outcomes. Most of the studies used inadequate research designs. Overall, positive training effects on the physicians' communication behaviour are found on half or less of the observed behaviours. Studies with the most adequate designs report the fewest positive training effects. CONCLUSION: Several reasons are discussed to explain the limited findings. Future research may benefit from research methods which focus on factors that inhibit and facilitate the physicians' implementation of skills into actual behaviours in daily practice. PMID- 10476017 TI - Medical students in ENT outpatient clinics: appointment times, patient satisfaction and student satisfaction. AB - OBJECTIVES: Outpatient clinics are increasingly important in medical education. The effect of students on clinic times and patient satisfaction, as well as their own satisfaction, were studied. DESIGN: A prospective, non-randomized, controlled study using adult patient questionnaires, medical student questionnaires and clinic time sheets. SETTING: Two teaching hospital ENT clinics. SUBJECTS: Medical students and adult patients. RESULTS: Three hundred and twenty-five patient questionnaires were collected (77% response), including 135 student encounters. Students did not affect appointment durations (19 min +/- 0.48 (standard error)) except at centre B (35 min +/- 1.1, P < 0.0001) where patient numbers were cut for teaching. Patient satisfaction, generally high, was not affected by students, appointment duration or gender of doctor or patient. It was slightly higher in the lower social classes (rs = 0.20, P = 0. 003) and older patients (rs = 0.17, P = 0.002). Student acceptability scores were not affected by student numbers (up to four), social class or time spent alone with students. They were higher if time was spent alone with the doctor (75.3% +/- 4.9) than not (63.0% +/- 1.8, P = 0.024). Thirty-six per cent of patients preferred to have a student present; only 9% preferred not. Student satisfaction was higher at centre B (73.7% +/- 2.3) where appointments were longer and students spent more time alone with patients than centre A (64.3% +/- 2.3, P = 0.0052). CONCLUSIONS: Clinic appointments are not necessarily longer in the presence of students. When students have the chance to see patients alone during longer consultations, student satisfaction is higher. Patient satisfaction, generally high, is not altered by the presence of students, but patients given time alone with their doctor are more accepting of students. These findings have resource implications for the planning of NHS clinics in teaching hospitals. PMID- 10476018 TI - Patients teach students: partners in arthritis education. AB - CONTEXT: A large metropolitan teaching hospital within The Northern Clinical School, University of Sydney. OBJECTIVE: To assess whether students taught by trained patients (Patient Partners) acquire the same levels of competence in musculoskeletal examination skills for arthritis as students taught by Consultant Rheumatologists. SUBJECTS: Year four medical students in a six-year Undergraduate Medical Programme. METHOD: Students randomized to eight tutorial groups were taught musculoskeletal examination skills in a 75-90 minute tutorial. Four groups were taught by Consultants with an untrained patient present and four groups were taught by Patient Partners. RESULTS: Students' mean self-ratings of skill before and after their tutorial were summed. For both groups, self-ratings before the tutorial were similar. After the tutorial both groups showed substantial gains in levels of skill. Patient Partners' ratings of students' taught by either Consultants or Partners were comparable. CONCLUSIONS: Patient Partners are at least equal to Consultant Rheumatologists in the teaching of musculoskeletal examination techniques for arthritis. PMID- 10476019 TI - Patient-centred consultations: a comparison of student experience and understanding in two clinical environments. AB - OBJECTIVES: To explore students' perspectives on doctor-patient communication. DESIGN: We interviewed students in focus groups before and after their placements. In semistructured interviews they were asked about their experiences of learning through clinical contact, with particular emphasis on patient-centred consultations, on the wards and during their community attachment. Students whose clinical skills training remained hospital-based were also interviewed to compare the range of experiences. SETTING: Leeds University. SUBJECTS: Third-year medical students. RESULTS: During hospital training students are rarely introduced, either through teaching or observation, to the idea of patient-centred consultations. In contrast both the principles underpinning and/or the practice of such consultations were routinely encountered during the general practice placement. Students considered the community attachment to have helped them appreciate the value of exploring patients' concerns both in terms of enhancing the patients' overall healthcare experience and promoting effective clinical management. While they considered the nature and purpose of hospital care to be qualitatively different from that conducted in the community, the relevance of adopting a patient-centred approach on the wards was confirmed. CONCLUSIONS: Recommendations for clinical training are made. The relevance and best use of community and hospital-based attachments need to be evaluated further. PMID- 10476020 TI - Students conducting consultations in general practice and the acceptability to patients. AB - OBJECTIVES: The General Medical Council has recommended that medical students should gain more experience in general practice. The study set out to determine patients' reactions to consultations conducted by a medical student alone prior to seeing their GP. DESIGN: A random sample of patients attending general practice surgeries in the Oxford area completed a questionnaire following consultation with a medical student. SETTING: Six general practice teaching surgeries. SUBJECTS: Fifth-year medical students. RESULTS: Of 130 responders 98% experienced no disadvantage in seeing the student; 35% considered that there were advantages in seeing the student; 98% said that they would be prepared to consult with a student again; 85% expressed no concerns about the gender of the student. CONCLUSIONS: The results of this study are very reassuring concerning the acceptability to patients of consulting with medical students and are more favourable than those reported for studies of students being present in consultations by GPs. PMID- 10476021 TI - Patients as teachers: a qualitative study of patients' views on their role in a community-based undergraduate project. AB - OBJECTIVES: Patients have been used in clinical medical education for many years with, traditionally, a relatively passive role. Following the General Medical Council recommendations for curricular change and the development of more community-based teaching, 'ordinary patients' in the community are increasingly being partnered with undergraduate students for particular projects. Very little research has been undertaken on patients' perceptions of this role. DESIGN: Semi structured interviews were carried out with 20 people to explore the views of patients taking part in a community-based undergraduate medical student project (the 'patient study') at Newcastle Medical School about their role as teachers of medical students, what they felt they had gained from participating, any problems or concerns and suggestions for change or improvement. SETTING: Newcastle Medical School, UK. SUBJECTS: Second-year medical students. RESULTS: Two major themes emerged. First, patients saw themselves in active roles as teachers: as experts in their medical condition; as exemplars of their condition; and as facilitators of the development of students' professional skills and attitudes. Secondly, patients felt they had benefited from participation, through talking about their problems; learning more about themselves; the satisfaction of helping; and from receiving gifts. In addition, a number of other issues were identified including interpersonal dynamics, gender and ethnic differences, inadequate briefing of participants and whether such community-based patient involvement might, in some situations, be felt to be exploitative. CONCLUSIONS: The study has shown that patients see themselves clearly as having specific contributions to make to medical students' education and training. This has implications for the further development of community-based teaching. PMID- 10476022 TI - Parents-as-teachers: design and establishment of a training programme for paediatric residents. AB - OBJECTIVES: To improve the training of paediatric residents in general and to provide more meaningful insights into family functioning. DESIGN: The programme is one facet of a 1-month developmental disabilities rotation for paediatric PL2 residents. It incorporates elements of family-centred care whereby physicians learn through home and community site visits about the importance of developing partnerships with parents and service providers in order to give better care to children with chronic conditions. SETTING: Gillette Specialty Health Care Center, St Paul, Minnesota. SUBJECTS: Paediatric PL2 residents and a parent group. RESULTS: Of 18 residents, 11 completed written and 18 completed oral feedback evaluations. There were infrequent but consistent complaints relating to logistics; more positive comments were made about the programme and individuals involved. CONCLUSIONS: In the minds of all participants, the Parents-as-Teachers (PAT) programme has been highly successful, as documented by short-term objective and subjective parent and resident feedback. The PAT experience represents a unique opportunity for parents and physicians-in-training to improve their mutual understanding. While not analysed in this report, the inclusion of a complementary programme involving on-site teaching in various community agencies also appears to be of value. PMID- 10476023 TI - Adolescent girls as simulators of medical illness. PMID- 10476024 TI - Modern medicine and the pursuit of cure. PMID- 10476025 TI - The plasmid prophage N15: a linear DNA with covalently closed ends. AB - Coliphage N15 is a temperate bacteriophage whose prophage is a linear plasmid molecule with covalently closed ends (telomeres). The N15 prophage provided the first example of such DNA in prokaryotes and, up to now, it is the only known example of a linear plasmid in Escherichia coli. The linear N15 mature phage DNA has single-stranded cohesive ends. The phage and plasmid prophage DNAs are circularly permuted. The nucleotide structure of the telomere-forming site tel RL in phage DNA corresponds to the structures of the terminal hairpin loops. It suggests a unique mechanism for conversion of the circular phage DNA to the linear plasmid form, which is performed by the prokaryotic telomerase (protelomerase). The results of a comparison of the protelomerase with integrases lead us to suggest that these proteins may have evolved from a common ancestor. The mechanism of plasmid N15 replication is unknown. We propose that the protelomerase participates in linear plasmid replication, acting as a resolvase of replicative intermediates that are tail-to-tail linear dimers. The sequence analysis of the N15 DNA showed that it represents an evolutionary 'link' between plasmids F, P1, P4 and lambdoid bacteriophages. PMID- 10476026 TI - Novel sensing mechanisms and targets for the cAMP-protein kinase A pathway in the yeast Saccharomyces cerevisiae. AB - The cAMP-protein kinase A (PKA) pathway in the yeast Saccharomyces cerevisiae plays a major role in the control of metabolism, stress resistance and proliferation, in particular in connection with the available nutrient conditions. Extensive information has been obtained on the core section of the pathway, i.e. Cdc25, Ras, adenylate cyclase, PKA, and on components interacting directly with this core section, such as the Ira proteins, Cap/Srv2 and the two cAMP phosphodiesterases. Recent work has now started to reveal upstream regulatory components and downstream targets of the pathway. A G-protein-coupled receptor system (Gpr1-Gpa2) acts upstream of adenylate cyclase and is required for glucose activation of cAMP synthesis in concert with a glucose phosphorylation-dependent mechanism. Although a genuine signalling role for the Ras proteins remains unclear, they appear to mediate at least part of the potent stimulation of cAMP synthesis by intracellular acidification. Recently, several new targets of the PKA pathway have been discovered. These include the Msn2 and Msn4 transcription factors mediating part of the induction of STRE-controlled genes by a variety of stress conditions, the Rim15 protein kinase involved in stationary phase induction of a similar set of genes and the Pde1 low-affinity cAMP phosphodiesterase, which specifically controls agonist-induced cAMP signalling. A major issue that remains to be resolved is the precise connection between the cAMP-PKA pathway and other nutrient-regulated components involved in the control of growth and of phenotypic characteristics correlated with growth, such as the Sch9 and Yak1 protein kinases. Cln3 appears to play a crucial role in the connection between the availability of certain nutrients and Cdc28 kinase activity, but it remains to be clarified which nutrient-controlled pathways control Cln3 levels. PMID- 10476027 TI - Molecular switches--the ON and OFF of bacterial phase variation. AB - The expression of most bacterial genes is controlled at the level of transcription via promoter control mechanisms that permit a graded response. However, an increasing number of bacterial genes are found to exhibit an 'all-or none' control mechanism that adapts the bacterium to more than one environment. One such mechanism is phase variation, traditionally defined as the high frequency ON<-->OFF switching of phenotype expression. Phase variation events are usually random, but may be modulated by environmental conditions. The mechanisms of phase variation events and their significance within the microbial community are discussed here. PMID- 10476028 TI - Cloning of the mspA gene encoding a porin from Mycobacterium smegmatis. AB - Porins form channels in the mycolic acid layer of mycobacteria and thereby control access of hydrophilic molecules to the cell. We purified a 100 kDa protein from Mycobacterium smegmatis and demonstrated its channel-forming activity by reconstitution in planar lipid bilayers. The mspA gene encodes a mature protein of 184 amino acids and an N-terminal signal sequence. MALDI mass spectrometry of the purified porin revealed a mass of 19 406 Da, in agreement with the predicted mass of mature MspA. Dissociation of the porin by boiling in 80% dimethyl sulphoxide yielded the MspA monomer, which did not form channels any more. Escherichia coli cells expressing the mspA gene produced the MspA monomer and a 100 kDa protein, which had the same channel-forming activity as whole-cell extracts of M. smegmatis with organic solvents. These proteins were specifically detected by a polyclonal antiserum that was raised to purified MspA of M. smegmatis. These results demonstrate that the mspA gene encodes a protein of M. smegmatis, which assembles to an extremely stable oligomer with high channel forming activity. Database searches did not reveal significant similarities to any other known protein. Southern blots showed that the chromosomes of fast growing mycobacterial species contain homologous sequences to mspA, whereas no hybridization could be detected with DNA from slow growing mycobacteria. These results suggest that MspA is the prototype of a new class of channel-forming proteins. PMID- 10476029 TI - Inactivation of the gene (cpe) encoding Clostridium perfringens enterotoxin eliminates the ability of two cpe-positive C. perfringens type A human gastrointestinal disease isolates to affect rabbit ileal loops. AB - Previous epidemiological studies have implicated Clostridium perfringens enterotoxin (CPE) as a virulence factor in the pathogenesis of several gastrointestinal (GI) illnesses caused by C. perfringens type A isolates, including C. perfringens type A food poisoning and non-food-borne GI illnesses, such as antibiotic-associated diarrhoea and sporadic diarrhoea. To further evaluate the importance of CPE in the pathogenesis of these GI diseases, allelic exchange was used to construct cpe knock-out mutants in both SM101 (a derivative of a C. perfringens type A food poisoning isolate carrying a chromosomal cpe gene) and F4969 (a C. perfringens type A non-food-borne GI disease isolate carrying a plasmid-borne cpe gene). Western blot analyses confirmed that neither cpe knock-out mutant could express CPE during either sporulation or vegetative growth, and that this lack of CPE expression could be complemented by transforming these mutants with a recombinant plasmid carrying the wild-type cpe gene. When the virulence of the wild-type, mutant and complementing strains were compared in a rabbit ileal loop model, sporulating (but not vegetative) culture lysates of the wild-type isolates induced significant ileal loop fluid accumulation and intestinal histopathological damage, but neither sporulating nor vegetative culture lysates of the cpe knock-out mutants induced these intestinal effects. However, full sporulation-associated virulence could be restored by complementing these cpe knock-out mutants with a recombinant plasmid carrying the wild-type cpe gene, which confirms that the observed loss of virulence for the cpe knock-out mutants results from the specific inactivation of the cpe gene and the resultant loss of CPE expression. Therefore, in vivo analysis of our isogenic cpe mutants indicates that CPE expression is necessary for these two cpe-positive C. perfringens type A human disease isolates to cause GI effects in the culture lysate:ileal loop model system, a finding that supports CPE as an important virulence factor in GI diseases involving cpe-positive C. perfringens type A isolates. PMID- 10476030 TI - Delayed nucleoid segregation in Escherichia coli. AB - To study the role of cell division in the process of nucleoid segregation, we measured the DNA content of individual nucleoids in isogenic Escherichia coli cell division mutants by image cytometry. In pbpB(Ts) and ftsZ strains growing as filaments at 42 degrees C, nucleoids contained, on average, more than two chromosome equivalents compared with 1.6 in wild-type cells. Because similar results were obtained with a pbpB recA strain, the increased DNA content cannot be ascribed to the occurrence of chromosome dimers. From the determination of the amount of DNA per cell and per individual nucleoid after rifampicin inhibition, we estimated the C and D periods (duration of a round of replication and time between termination and cell division respectively), as well as the D' period (time between termination and nucleoid separation). Compared with the parent strain and in contrast to ftsQ, ftsA and ftsZ mutants, pbpB(Ts) cells growing at the permissive temperature (28 degrees C) showed a long D' period (42 min versus 18 min in the parent) indicative of an extended segregation time. The results indicate that a defective cell division protein such as PbpB not only affects the division process but also plays a role in the last stage of DNA segregation. We propose that PbpB is involved in the assembly of the divisome and that this structure enhances nucleoid segregation. PMID- 10476031 TI - Insertion of a Yop translocation pore into the macrophage plasma membrane by Yersinia enterocolitica: requirement for translocators YopB and YopD, but not LcrG. AB - The Yersinia survival strategy is based on its ability to inject effector Yops into the cytosol of host cells. Translocation of these effectors across the eukaryotic cell membrane requires YopB, YopD and LcrG, but the mechanism is unclear. An effector polymutant of Y. pseudotuberculosis has a YopB-dependent contact haemolytic activity, indicating that YopB participates in the formation of a pore in the cell membrane. Here, we have investigated the formation of such a pore in the plasma membrane of macrophages. Infection of PU5-1.8 macrophages with an effector polymutant Y. enterocolitica led to complete flattening of the cells, similar to treatment with the pore-forming streptolysin O from Streptococcus pyogenes. Upon infection, cells released the low-molecular-weight marker BCECF (623 Da) but not the high-molecular-weight lactate dehydrogenase, indicating that there was no membrane lysis but, rather, insertion of a pore of small size into the macrophage plasma membrane. Permeation to lucifer yellow CH (443 Da) but not to Texas red-X phalloidin (1490 Da) supported this hypothesis. All these events were found to be dependent not only on translocator YopB as expected but also on YopD, which was required equally. In contrast, LcrG was not necessary. Consistently, lysis of sheep erythrocytes was also dependent on YopB and YopD, but not on LcrG. PMID- 10476032 TI - Stimulation of transposition of the Mycobacterium tuberculosis insertion sequence IS6110 by exposure to a microaerobic environment. AB - The Mycobacterium tuberculosis-specific insertion sequence IS6110/986 has been widely used as a probe because of the multiple polymorphism observed among different strains. To investigate transposition of IS6110, a series of artificially constructed composite transposons containing IS6110 and a kanamycin resistance marker were constructed. The composite transposons were inserted into a conditionally replicating, thermosensitive, Escherichia coli-mycobacterial shuttle vector and introduced into M. smegmatis mc2155. Lawns of transformants were grown at the permissive temperature on kanamycin-supplemented agar and subsequently prevented from further growth by shifting to the non-permissive temperature. Under normal atmospheric conditions, kanamycin-resistant papillae appeared after only about 5-6 weeks of incubation. However, these events were not associated with transposon mobilization. In contrast, lawns that were exposed to a 48 h microaerobic shock generated kanamycin-resistant papillae after only 6-14 days. These events were generated by conservative transposition of the IS6110 composite transposon into the M. smegmatis chromosome, with loss of the shuttle vector. In common with other IS3 family elements, transposition of IS6110 is thought to be controlled by translational frameshifting. However, we were unable to detect any significant frameshifting within the putative frameshifting site of IS6110, and the level of frameshifting was not affected by microaerobic incubation. The finding that transposition of IS6110 is stimulated by incubation at reduced oxygen tensions may be relevant to transposition of IS6110 in M. tuberculosis harboured within TB lesions. PMID- 10476033 TI - Ambient pH signal transduction in Aspergillus: completion of gene characterization. AB - Completing the molecular analysis of the six pal genes of the ambient pH signal transduction pathway in Aspergillus nidulans, we report the characterization of palC and palH. The derived translation product of palH contains 760 amino acids with prediction of seven transmembrane domains in its N-terminal moiety. Remarkably, a palH frameshift mutant lacking just over half the PalH protein, including almost all of the long hydrophilic region C-terminal to the transmembrane domains, retains some PalH function. The palC-derived translation product contains 507 amino acids, and the null phenotype of a frameshift mutation indicates that at least one of the C-terminal 142 residues is essential for function. Uniquely among the A. nidulans pH-signalling pal genes, palC appears to have no Saccharomyces cerevisiae homologue, although it does have a Neurospora crassa expressed sequence tag homologue. In agreement with findings for the palA, palB and palI genes of this signalling pathway, levels of the palC and palH mRNAs do not appear to be pH regulated. PMID- 10476034 TI - Sequence-selective interactions with RNA by CspB, CspC and CspE, members of the CspA family of Escherichia coli. AB - The CspA family of Escherichia coli comprises nine homologous proteins, CspA to CspI. CspA, the major cold shock protein, binds RNA with low sequence specificity and low binding affinity. This is considered to be important for its proposed function as an RNA chaperone to prevent the formation of secondary structures in RNA molecules, thus facilitating translation at low temperature. The cellular functions of other Csp proteins are yet to be fully elucidated, and their sequence specific binding capabilities have not been identified. As a step towards identification of the target genes of Csp proteins, we investigated the RNA binding specificities of CspB, CspC and CspE by an in vitro selection approach (SELEX). In the present study, we show that these proteins are able to bind preferentially to specific RNA/single-stranded DNA sequences. The consensus sequences for CspB, CspC and CspE are U/T stretches, AGGGAGGGA and AU/AT-rich regions, especially AAAUUU, respectively. CspE and CspB have Kd values in the range 0.23-0.9 x 10(-6) M, while CspC has 10-fold lower binding affinity. Consistent with our recent findings of transcriptional regulation of cspA by CspE, we have identified a motif identical to the CspE consensus. This motif is the putative CspE-mediated transcription pause recognition site in a 5' untranslated region of the cspA mRNA. PMID- 10476035 TI - Characterization of a morphological checkpoint coupling cell-specific transcription to septation in Bacillus subtilis. AB - Early in the process of spore formation in Bacillus subtilis, asymmetric cell division produces a large mother cell and a much smaller prespore. Differentiation of the prespore is initiated by activation of an RNA polymerase sigma factor, sigmaF, specifically in that cell. sigmaF is controlled by a regulatory cascade involving an anti-sigma factor, SpoIIAB, an anti-anti-sigma factor, SpoIIAA, and a membrane-bound phosphatase, SpoIIE, which converts the inactive, phosphorylated form of SpoIIAA back to the active form. SpoIIE is required for proper asymmetric division and much of the protein is sequestered into the prespore during septation. Importantly, activation of sigmaF is dependent on formation of the asymmetric septum. We have now characterized this morphological checkpoint in detail, using strains affected in cell division and/or spoIIE function. Surprisingly, we found that significant dephosphorylation of SpoIIAA occurred even in the absence of septation. This shows that the SpoIIE phosphatase is at least partially active independent of the morphological event and also that cells can tolerate significant levels of unphosphorylated SpoIIAA without activating sigmaF. We also describe a spoIIE mutant in which the checkpoint is bypassed, probably by an increase in the dephosphorylation of SpoIIAA. Taken together, the results support the idea that sequestration of SpoIIE protein into the prespore plays an important role in the control of sigmaF activation and in coupling this activation to septation. PMID- 10476036 TI - Norfloxacin-induced DNA cleavage occurs at the dif resolvase locus in Escherichia coli and is the result of interaction with topoisomerase IV. AB - The dif locus is a site-specific recombination site located within the terminus region of the chromosome of Escherichia coli. Recombination at dif resolves circular dimer chromosomes to monomers, and this recombination requires the XerC, XerD and FtsK proteins, as well as cell division. In order to characterize other enzymes that interact at dif, we tested whether quinolone-induced cleavage occurs at this site. Quinolone drugs, such as norfloxacin, inhibit the type 2 topoisomerases, DNA gyrase and topoisomerase IV, and can cleave DNA at sites where these enzymes interact with the chromosome. Using strains in which either DNA gyrase or topoisomerase IV, or both, were resistant to norfloxacin, we determined that specific interactions between dif and topoisomerase IV caused cleavage at that site. This interaction required XerC and XerD, but did not require the C-terminal region of FtsK or cell division. PMID- 10476037 TI - The beta-barrel domain of FhuADelta5-160 is sufficient for TonB-dependent FhuA activities of Escherichia coli. AB - FhuA in the outer membrane of Escherichia coli serves as a transporter for ferrichrome, the antibiotics albomycin and rifamycin CGP4832, colicin M, and as receptor for phages T1, T5 and phi80. The previously determined crystal structure reveals that residues 160-714 of the mature protein form a beta-barrel that is closed from the periplasmic side by the globular N-proximal fragment, residues 1 159, designated the cork. In this study, deletion of the cork resulted in a stable protein, FhuADelta5-160, that was incorporated in the outer membrane. Cells that synthesized FhuADelta5-160 displayed a higher sensitivity to large antibiotics such as erythromycin, rifamycin, bacitracin and vancomycin, and grew on maltotetraose and maltopentaose in the absence of LamB. Higher concentrations of ferrichrome supported growth of a tonB mutant that synthesized FhuADelta5-160. These results demonstrate non-specific diffusion of compounds across the outer membrane of cells that synthesize FhuADelta5-160. However, growth of a FhuADelta5 160 tonB wild-type strain occurred at low ferrichrome concentrations, and ferrichrome was transported at about 45% of the FhuA wild-type rate despite the lack of ferrichrome binding sites provided by the cork. FhuADelta5-160 conferred sensitivity to the phages and colicin M at levels similar to that of wild-type FhuA, and to albomycin and rifamycin CGP 4832. The activity of FhuADelta5-160 depended on TonB, although the mutant lacks the TonB box (residues 7-11) previously implicated in the interaction of FhuA with TonB. CCCP inhibited tonB dependent transport of ferrichrome through FhuADelta5-160. FhuADelta5-160 still functions as a specific transporter, and sites in addition to the TonB box are involved in the TonB-mediated response of FhuA to the proton gradient of the cytoplasmic membrane. It is proposed that TonB interacts with the TonB box of FhuA and with the beta-barrel to release ferrichrome from the FhuA binding sites and to open the channel in FhuA. For transport of ferrichrome through the open channel of FhuADelta5-160, interaction of TonB with the beta-barrel is sufficient to release ferrichrome from the residual binding sites at the beta-barrel and to induce the active conformation of the L4 loop at the cell surface for infection by the TonB-dependent phages T1 and phi80. PMID- 10476038 TI - A gene required for the regulation of photosynthetic light harvesting in the cyanobacterium Synechocystis 6803. AB - A gene required for the short-term regulation of photosynthetic light harvesting (the state transition) has been identified in the cyanobacterium Synechocystis sp. PCC6803. The open reading frame is designated sll1926 in the complete Synechocystis gene sequence. The deduced amino acid sequence has no homologues in current sequence databases and no recognizable sequence motifs. It encodes a putative integral membrane protein of 16 kDa, which we have designated RpaC (regulator of phycobilisome association C). Fluorescence measurements of an insertional inactivation mutant of rpaC (Deltasll1926) show that it is specifically unable to perform state transitions. Deltasll1926 has approximately wild-type levels of PS1, PS2 and phycobilisomes. Measurements of oxygen evolution and uptake show Deltasll1926 to have no deficiency in electron transport rates. In vitro [gamma-32P]-ATP labelling experiments suggest that RpaC is not the 15 kDa membrane phosphoprotein previously implicated in state transitions. Deltasll1926 grows more slowly than the wild type only at very low light intensities. PMID- 10476039 TI - Tn5053 family transposons are res site hunters sensing plasmidal res sites occupied by cognate resolvases. AB - DNA sequence database search revealed that most of Tn5053/Tn402 family transposons inserted into natural plasmids were located in putative res regions upstream of genes encoding various resolvase-like proteins. Some of these resolvase genes belonged to Tn3 family transposons and were closely related to the tnpR genes of Tn1721 and a recently detected Tn5044. Using recombinant plasmids containing fragments of Tn1721 or Tn5044 as targets in transposition experiments, we have demonstrated that Tn5053 displays striking insertional preference for the res regions of these transposons: more than 70% of Tn5053 insertion events occur in clusters inside the target res regions, while most remaining insertion events occur no further than 200 base pairs away from both sides of the res regions. We demonstrate that Tn5053 insertions (both into and outside a res region of the target plasmid) require the presence of a functional cognate resolvase gene either in cis or in trans. To our knowledge, this is the first case when a site-specific recombination system outside a transposon has been shown to be involved in transposition. PMID- 10476040 TI - Pseudomonas aeruginosa AlgZ, a ribbon-helix-helix DNA-binding protein, is essential for alginate synthesis and algD transcriptional activation. AB - The Pseudomonas aeruginosa algD gene is the first gene of an operon encoding most of the enzymes necessary for biosynthesis of the exopolysaccharide alginate. Transcriptional activation of algD results in the high-level synthesis of alginate, an important P. aeruginosa virulence factor with antiphagocytic and adherence properties. Previously, we have identified a protein(s), AlgZ, expressed in mucoid P. aeruginosa CF isolates that specifically bound to sequences located 280 bp upstream of the algD promoter. Mutagenesis of the AlgZ DNA binding site and transcription assays were used to show that AlgZ was an activator of algD transcription. In the current study, the monomeric size of AlgZ was estimated to be between 6 kDa and 15 kDa by electroelution of a protein preparation from an SDS-PAGE gel and analysis of the fractions via protein staining and electrophoretic mobility shift assays. A biochemical enrichment procedure, resulting in a 130-fold enrichment for AlgZ, was devised, the protein identified and a partial amino-terminal sequence obtained. Using the P. aeruginosa Genome Project database, a complete sequence was obtained, and algZ was cloned and expressed in Escherichia coli. Expression of algZ was sufficient for the observed AlgZ DNA binding previously observed from extracts of P. aeruginosa. A protein database search revealed that AlgZ is homologous to the Mnt and Arc repressors of the ribbon-helix-helix family of DNA-binding proteins. An algZ deletion mutant was constructed in the mucoid CF isolate FRD1. The resulting strain was non-mucoid and exhibited no detectable algD transcription. As an indirect role in transcription would probably result in some residual algD transcription, these data suggest that AlgZ is an integral activator of algD and support the hypothesis that both AlgZ and the response regulator AlgR are involved in direct contact with RNA polymerase containing the alternative sigma factor, AlgT. The cloning of algZ is a crucial step in determining the mechanism of algD activation. PMID- 10476041 TI - Expression and heat-responsive regulation of a TFIIB homologue from the archaeon Haloferax volcanii. AB - Multiple divergent genes encoding the eukaryal-like TFIIB (TFB) transcription initiation factor have been identified in the archaeon Haloferax volcanii. Expression of one of these TFB-encoding genes, referred to here as tfb2, was induced specifically in response to heat shock at the transcription level. A time course for tfb2 induction demonstrated that mRNA levels increased as much as eightfold after 15 min at 60 degrees C. A transcription fusion of the tfb2 promoter region with a stable RNA reporter gene confirmed the heat responsiveness of the tfb2 core promoter, and immunoblot analysis using antibodies generated against a recombinant His-tagged TFB2 showed that the protein levels of one TFB increased slightly in response to elevated temperatures. An archaeal consensus TATA element (5'-TTTATA-3') was located 110 bp upstream of the translation start site and appeared to be used for both basal and heat shock-induced expression. The long DNA leader region (79 bp) preceding the predicted AUG translation start codon for TFB2 contained a T-rich sequence element located 22 bp downstream of the transcription start site. Using an in vivo transcription termination assay, we demonstrated that this T-rich element can function as a sequence-dependent transcription terminator, which may serve to downregulate expression of the tfb2 gene under both non-heat shock and heat shock conditions. PMID- 10476042 TI - CADASIL: hereditary disease of arteries causing brain infarcts and dementia. AB - Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) begins with migraine with aura in approximately one third of the patients. More severe symptoms of recurrent strokes usually appear at 30-50 years of age. However, well before the first stroke, CADASIL may be diagnosed on the basis of characteristic hyperintensities in T2-weighted magnetic resonance images. Multiple lacunar infarcts located mainly in the basal ganglia and frontal white matter lead to a cognitive decline and finally to dementia. These infarcts result from a thickening and fibrosis of the walls of the small and medium-sized penetrating arteries with consequent obliteration and/or thrombosis. Although the symptoms are almost exclusively neurological, the arteriopathy is generalized. Thus, basophilic, periodic acid-Schiff-positive and, in electron microscopy, osmiophilic material accumulates between degenerating smooth muscle cells. This occurs even in dermal arteries, which renders skin a useful target for diagnostic biopsy. Presently, no specific therapy is available. CADASIL is caused by missense point mutations in the Notch3 gene, which encodes a transmembrane receptor protein. Each gene defect leads to either a gain or loss of a cysteine residue in the extracellular, N-terminal domain of the molecule, which most probably results in conformational alteration. The function of Notch3 in adults and the pathogenesis of CADASIL are still unknown. PMID- 10476043 TI - Laminar distribution of pick bodies, pick cells and Alzheimer disease pathology in the frontal and temporal cortex in Pick's disease. AB - Lesions in Alzheimer's disease (AD) and dementia with Lewy bodies (DLB) have distinct laminar distributions in the cortex. The objective of the present study was to test the hypothesis that the lesions characteristic of Pick's disease (PD) and AD have distinctly different laminar distributions in cases of PD. Hence, the laminar distribution of Pick bodies (PB), Pick cells (PC), senile plaques (SP) and neurofibrillary tangles (NFT) was studied in the frontal and temporal cortex in nine patients with PD. In 57% of analyses of individual cortical areas, the density of PB was maximal in the upper cortex while in 25% of analyses, the distribution of PB was bimodal with density peaks in the upper and lower cortex. The density of PC was maximal in the lower cortex in 77% of analyses while a bimodal distribution was present in 5% of analyses. The density of NFT was maximal in the upper cortex in 50% of analyses, in the lower cortex in 15% of analyses, with a bimodal distribution in 4% of analyses. The density of SP did not vary significantly with cortical depth in 86% of analyses. The vertical densities of PB and PC were negatively correlated in 12/21 (57%) of brain areas. The maximum density of PB in the upper cortex was positively correlated with the maximum density of PC in the lower cortex. In 17/25 (68%) of brain areas, there was no significant correlation between the vertical densities of PB and NFT. The data suggest that the pathogenesis of PB may be related to that of the PC. In addition, although in many areas PB and NFT occur predominantly in the upper cortex, the two lesions appeared to affect different neuronal populations. PMID- 10476044 TI - Cell death and oxidative stress in gliomas. AB - In gliomas, apoptosis and necrosis are determined by a number of promoting and inhibiting factors including oxidative cell stress mediated by nitric oxide synthases (NOS) and reduced by superoxide dismutases. Therefore, in 46 gliomas (including astrocytomas, oligodendrogliomas, oligo-astrocytomas, and glioblastomas), the relationship of apoptosis and necrosis and the expression of apoptosis-promoting (p53, bax, Fas, Fas-L) and inhibiting (bcl-2) factors as well as of different isoforms of NOS (NOSb, NOSe, NOSi) and manganese superoxide dismutase (MnSOD) were studied. Apoptosis was measured in situ by the TUNEL method while expression profiles of apoptosis-related and oxidative stress associated factors were determined by immunohistochemistry. As a defining criterion, necrosis was restricted to glioblastomas while apoptosis increased with tumour malignancy (P=0.017) in all types of gliomas. Glial tumour cells displayed upregulation of bax, Fas, Fas-L, p53, and bcl-2 but with no significant correlation with malignancy. There was also a strong expression of NOS isoforms with upregulation of NOSe in all and of NOSb and NOSi in nearly 50% of the tumour specimens but only NOSb expression correlated significantly with tumour malignancy (P=0.004). Likewise, MnSOD was strongly expressed in all gliomas but was not correlated with tumour grade. There was a wide variability of expression in each tumour type without significant correlation between apoptosis and expression of apoptosis-associated or oxidative stress-related factors indicating that the network of regulating factors may be too complex for clear associations. PMID- 10476045 TI - Morphological correlates of neurological dysfunction in macaques infected with neurovirulent simian immunodeficiency virus. AB - The pattern of neurological disease caused by human immunodeficiency virus (HIV) infection of the central nervous system (CNS) was investigated using a macaque model of acquired immune defiency syndrome (AIDS). Seven of nine macaques inoculated with neurovirulent simian imunodeficiency virus (SIVmac ) developed AIDS within 3 months. Four of these had clinically obvious neurological disease and extensive conduction defects in the form of latency increases in evoked potential (EP) responses. Neuropathologically, all four animals had disseminated white matter disease in the form of multifocal, perivascular and nodular parenchymal mononuclear cell infiltrates, along with extensive involvement of the cortical grey matter, leptomeninges and intracranial portions of cranial nerves. A brisk multinucleated giant cell (MGC) response was a frequent accompaniment in the affected areas. Three of the animals in this group also showed spongiform vacuolation in the occipital grey matter, a lesion described only rarely in HIV encephalitis. In the remaining three animals, there was only minimal evidence of overt neurological impairment or conduction defects. These animals had only mild to moderate neuropathological changes and lesions were virtually confined to the white matter regions of the brain. MGC responses were rare or absent in the CNS of these animals. Neuropathological findings in this SIVmac model have therefore shown good correlation with the severity of clinical and neurophysiological changes, and are reminiscent of HIV-1 encephalitis. More importantly, white matter involvement was a consistent finding in the affected macaques, regardless of the duration and severity of disease, or type of virus inoculated, suggesting an unusual susceptibility for lentiviral infection in these regions of the macaque CNS. PMID- 10476046 TI - Low amyloid (Abeta) plaque load and relative predominance of diffuse plaques distinguish argyrophilic grain disease from Alzheimer's disease. AB - Argyrophilic grain disease constitutes one cause of late-onset dementia. Its classification among dementia disorders is still unclear because most of the reported argyrophilic grain disease cases are associated with neurofibrillary lesions (e.g. neurofibrillary tangles) which are also typical of Alzheimer's disease. In the present study we determine whether argyrophilic grain disease is associated with the senile plaques of Alzheimer's disease. The distribution and density of senile plaques was systematically investigated in 11 demented argyrophilic grain disease cases using Abeta immunohistochemistry and stereological techniques, and the results were compared with 11 Alzheimer's disease cases. All subjects with argyrophilic grain disease exhibited neurofibrillary changes corresponding to Braak stages I-III. Three of the 11 argyrophilic grain disease cases (27%) were completely devoid of Abeta deposits. In argyrophilic grain disease cases with senile plaques, the average total plaque load was significantly lower (1%) than in Alzheimer's disease (3.1%) (P<0. 005). The regional distribution of the senile plaques and the proportion of diffuse vs. primitive or mature plaques in argyrophilic grain disease resembled values of senile plaques reported in non-demented elderly subjects, and was significantly different from Alzheimer's disease. Similarly the immunocytochemical profile of the Abeta deposition in argyrophilic grain disease resembled that of non-demented elderly subjects rather than that of subjects with Alzheimer's disease. As all argyrophilic grain disease cases under investigation were demented, including those devoid of senile plaques, the present study further supports the thesis that dementia in argyrophilic grain disease correlates more with the density and distribution of argyrophilic grains than with associated lesions of the Alzheimer type. PMID- 10476047 TI - Ultrastructural changes in peripheral nerve in hereditary motor and sensory neuropathy-Lom. AB - Ultrastructural observations have been made on nerve biopsy specimens from five cases of hereditary motor and sensory neuropathy-Lom (HMSNL). A number of features that distinguish it from other hereditary demyelinating neuropathies were identified. Teased fibre studies were not feasible but examination of longitudinal sections by electron microscopy demonstrated demyelination/remyelination. Severe progressive axonal loss was a conspicuous feature. There was no indication of axonal atrophy. Hypertrophic onion bulb changes were present in the younger patients which later regressed, probably secondary to axonal loss. Myelin thickness was generally reduced in relation to axon diameter, indicating hypomyelination, and partial ensheathment of axons by Schwann cells was observed. The Schmidt-Lanterman incisures were atypical in extending for long lengths along the internode. Uncompacted myelin with a periodicity greater than that observed in other neuropathies in which it occurs was a feature, as was the accumulation of pleomorphic material in the adaxonal Schwann cell cytoplasm. An unusual finding was the presence of intra-axonal accumulations of irregularly arranged curvilinear profiles. These resemble those that have been described in experimental vitamin E deficiency. The amount of endoneurial collagen was markedly increased and some endoneurial blood vessels showed a non-specific basal laminal reduplication. PMID- 10476048 TI - CD44 expression in primary and recurrent oligodendrogliomas and in adjacent gliotic brain tissue. AB - CD44 expression was evaluated in 114 primary and recurrent oligodendrogliomas (46 primary oligodendrogliomas grade II and 15 recurrences grade II; 17 primary anaplastic oligodendrogliomas and nine recurrent oligodendrogliomas grade III; 14 glioblastomas with oligodendroglial growth pattern and 13 tumour recurrences grade IV). CD44 expression was found to correlate with tumour grading (P<0.001) and with survival (Kaplan-Meier Log Rank P<0.01, median survival 28 months in oligodendrogliomas with CD44 expression vs. 108 months in CD44-negative tumours). However, multivariate Cox regression analysis of grading and CD44 expression revealed that CD44 expression had no prognostic relevance independent of histological grading. Characterization of CD44 positive cells by double labelling with GFAP revealed that in addition to oligodendroglioma cells, reactive astrocytes within the tumour, at the invasive margin and along the pathways of oligodendroglioma invasion in the subpial matrix, and in the vicinity of vessels, frequently expressed CD44. It is suggested that in analogy to carcinoma invasion where a tumour-induced production of hyaluronan was found in fibroblasts at the invasive margin of the tumour, in the brain reactive astrocytes may produce hyaluronan which would facilitate the adhesion of new CD44-positive astrocytic processes but which would also promote tumour invasion. PMID- 10476049 TI - Galectin-3 and galectin-3-binding site expression in human adult astrocytic tumours and related angiogenesis. AB - Using computer-assisted microscopy, the present work aimed to quantitatively characterize the level of the histochemically detectable expression of galectin-3 and galectin-3-binding sites in sections of a series of 84 astrocytic tumours (including 22 grade II, 21 grade III and 41 grade IV specimens) and seven non tumoural specimens used as controls. The presence of galectin-3 and reactive sites for this lectin were monitored by means of a specific polyclonal anti galectin-3 antibody (aGal3) and biotinylated galectin-3 (Gal3), respectively. The pattern of expression of galectin-3-binding sites is compared to the pattern of expression of laminin (a potential galectin-3 ligand) revealed using a biotinylated anti-laminin antibody (aLam). Three variables quantitatively characterizing histochemical staining reactions were evaluated by means of computer-assisted microscopy for each of the 3 probes under study (aGal3, Gal3 and aLam). The labelling index (LI) is the percentage of tissue area specifically stained by a histochemical probe. The mean optical density (MOD) denotes staining intensity. The concentration heterogeneity (CH) feature expresses the concentrational spread of individual fields. The data obtained in the present study show that: (i) white matter of a non-tumoural brain expresses galectin-3 (and also galectin-3-binding sites); (ii) the level of galectin-3 expression significantly decreases in the majority of tumour astrocytes from low to high grade astrocytic tumours; while (iii) some tumour cell clones expressing high amounts of galectin-3 emerged with increasing levels of malignancy; and (iv) the level of accessible galectin-3-binding sites was apparently not heavily modified in the course of malignancy progression. In conclusion, the results obtained in the present study show that human astrocytic tumours are very heterogenous in their galectin-3 levels of expression. If high levels of galectin-3 determine the invasiveness potential of a tumour cell, then within a heterogenous tumour the presence of even a small, but actively proliferating number of tumour cell clones expressing high levels of galectin-3 can be expected to lead to tumour invasiveness. PMID- 10476050 TI - Evidence of blood-brain barrier dysfunction in human cerebral malaria. AB - Patients infected with the malaria parasite Plasmodium falciparum may develop a diffuse reversible encephalopathy, termed cerebral malaria. It is unclear how the intraerythrocytic parasite, which sequesters in the cerebral microvasculature but does not enter the brain parenchyma, induces this neurological syndrome. Adhesion of parasitized red blood cells in the brain microvasculature is mediated by specific receptors on the host endothelium, including intercellular adhesion molecule (ICAM)-1, CD36 and CD31. Leucocyte binding to cerebral endothelial cells in culture induces intracellular signalling via ICAM-1. The hypothesis that parasitized red blood cells binding to receptors on cerebral endothelial cells causes changes in the integrity of the blood-brain barrier was tested. Immunohistochemistry was used to examine the blood-brain barrier in human cerebral malaria, with antibodies to macrophage and endothelial activation markers, intercellular junction proteins, and plasma proteins. The distribution of the cell junction proteins occludin, vinculin and ZO-1 were altered in cerebral malaria cases compared to controls. While fibrinogen was the only plasma protein detected in the perivascular space, there was widespread perivascular macrophage activation, suggesting that these cells had been exposed to plasma proteins. It was concluded that functional changes to the blood-brain barrier occur in cerebral malaria, possibly as a result of the binding of parasitized red blood cells to cerebral endothelial cells. These changes require further examination in vitro. PMID- 10476051 TI - Intracranial cavity volume can be accurately estimated from the weights of intracranial contents: confirmation by the dental plaster casting method. AB - Intracranial cavity volume is used to evaluate brain size relative to the intracranial space. This volume can theoretically be obtained from the weights and densities of the brain and surrounding cerebrospinal fluid (weight method). However, the accuracy of this method has not been examined. In this study, we examined the reliability of the weight method, by comparing the intracranial cavity volumes of 41 post-mortem cases obtained by the weight method (ICVw) with those obtained by a dental plaster casting method (ICVcast) which was shown to be unbiased. The ICVw was not significantly different from the ICV cast (P=0.49, paired t-test), and the standard error of difference was 18 ml (1.3% of ICVcast). These results show that the weight method is reliable, and applicable to routine autopsies. PMID- 10476052 TI - Leptomeningeal melanoma and Creutzfeldt-Jakob disease in a patient with chronic lymphocytic leukaemia. AB - A 78-year-old woman with known chronic lymphocytic leukaemia (CLL) was admitted to a psychiatric unit because of rapidly declining cognitive function. Clinical examination also revealed cerebellar signs and she later became akinetic and mute. She deteriorated and died of bronchopneumonia. The histology from the post mortem confirmed the presence of CLL in the lymph nodes and she was also found to have diffuse leptomeningeal melanoma. In addition, there was extensive prion protein deposition in the cerebral cortex, but without significant spongiosis. The astrocytosis that was present appeared superficial only. Furthermore, prion protein appeared to be co-expressed with betaA4 in the form of plaques. The patient therefore had evidence of sporadic Creutzfeldt-Jakob disease (CJD) in addition to meningeal melanoma and CLL. This case further illustrates the importance of employing prion protein immunohistochemistry in suspected cases of CJD, especially where the histology is atypical. PMID- 10476053 TI - A hookworm allergen which strongly resembles calreticulin. AB - Immmoglobulin E-rich plasma from patients from Papua New Guinea infected with Necator americanus has been used to probe an adult N. americanus cDNA library for the presence of hookworm allergens. Using this approach, one hookworm allergen has been identified as calreticulin, which was subsequently expressed in Escherichia coli. Little serological cross reactivity was seen between the recombinant calreticulins of this hookworm and its host. Prospective roles for hookworm calreticulin in the host-parasite relationship are discussed in depth. PMID- 10476054 TI - Local and systemic cytokine expression during experimental chronic Trypanosoma cruzi infection in a Cebus monkey model. AB - Cebus apella is an acceptable model for chronic chagasic cardiomyopathy (CCC), since it is possible to experimentally induce cardiac lesions after 1 year of Trypanosoma cruzi infection. The T. cruzi Y strain, shown previously to produce CCC in C. apella monkeys, was used to experimentally infect 10 monkeys. Parasitological, serological and clinical parameters were monitored during a 19 month follow-up, and systemic cytokine responses were assessed sequentially in five monkeys selected according to the differential parasitemia pattern exhibited. Ten additional monkeys, infected with the same strain for 5, 10 and 12 years, were analysed cross-sectionally. Three monkeys/time point and one uninfected control animal were sacrificed for gross pathology, histology, presence of parasites, and local cytokine gene expression. Elevated expression of interleukin (IL)-4 was observed throughout the study in monkeys that had persistent, high parasitemias, whereas a high level of interferon (IFN)-gamma was seen in monkeys that controlled parasitemias soon after infection. Chronically infected monkeys expressed a nonpolarized, Th0-type response. Cardiac tissue collected from a monkey that succumbed to acute infection had elevated levels of proinflammatory cytokine [IL-1beta, IL-6, tumour necrosis factor-alpha] and interstitial cell adhesion molecule (ICAM)-1, platelet-derived growth factor (PDGF)-alpha, transforming growth factor (TGF)-beta and IL-10 transcripts. Cytokine production in cardiac tissue of chronically infected monkeys was also characterized by elevated expression of ICAM-1, PDGF-alpha and TGF-beta, which correlated with the detection of T. cruzi DNA by polymerase chain reaction. PMID- 10476055 TI - Vaccination against Leishmania major in a CBA mouse model of infection: role of adjuvants and mechanism of protection. AB - Gp63 is a major surface protein of Leishmania promastigotes. Its protective efficacy has been tested in several experimental models using different mouse strains, gp63 forms, adjuvants and routes of immunization, giving rise to conflicting results. This investigation was designed to determine whether these discrepancies could be ascribed to differing experimental procedures, and to compare gp63-induced protection with that achieved using live promastigotes. Preliminary experiments demonstrated that gp63 was an extremely potent immunogen compared to a standard antigen (ovalbumin). Protection against Leishmania major infection afforded by gp63 inoculation was studied in CBA mice. Injection of gp63 in saline, or of CFA, BCG, and C. parvum without antigen, induced significant protection. When gp63 and adjuvants were combined, results differed depending on the site of vaccination relative to that of the challenge infection. Vaccination with gp63 plus adjuvants in the tail (i.e. close to the site of infection) led to a stronger reduction of lesion size than the basal level of protection elicited by adjuvants alone, except in the case of CFA. Surprisingly however, when the antigen was injected at a distance from the site of infection (immunization in the hind foot pads, infection in the rump), the protective effect of gp63 was decreased by the adjuvants. Finally, vaccination at either site using live parasites (radioattenuated or virulent promastigotes) resulted in most instances in better protection than achieved by any protocol using gp63 and adjuvants. While anti-gp63 T cells proliferated in vitro in response to L. major-infected bone marrow-derived macrophages, they were unable to activate macrophages for parasite killing. This is in contrast with lymphocytes from mice immunized with live parasites, which both proliferated and stimulated significant killing of the microorganisms within 48 h. PMID- 10476056 TI - Secondary alveolar echinococcosis in lymphotoxin-alpha and tumour necrosis factor alpha deficient mice: exacerbation of Echinococcus multilocularis larval growth is associated with cellular changes in the periparasitic granuloma. AB - The availability of mice carrying a deletion of LT-alpha and tumour necrosis factor (TNF)-alpha genes enabled us to investigate the role of the TNF during alveolar echinococcosis. We compared the growth rate of Echinococcus multilocularis in LT-alphaTNF-alpha +/+ mice to that of mice having either no or only one LT-alphaTNF-alpha functionnal allele. LT-alphaTNF-alpha -/- mice harboured a significantly higher parasite burden than did the other two populations at 5, 10, and 15 weeks of infection, and they did not survive thereafter. Liver metacestodes removed from these mice were alive and the dehydrogenase activities of peritoneal metacestodes were decreased. Liver lesions regressed in most wild-type mice. Indeed, dead parasites were cordoned by granulomas containing numerous macrophages and lymphocytes leading to focal liver fibrosis at an early stage of infection. In contrast, most of LT-alphaTNF-alpha /- mice harboured metacestodes interspersed with leucocytes, realising purulent abscesses with secondary extensive irregular fibrosis at a late stage of infection. Heterozygous mice had behavioural characteristics intermediate between homozygous mutants and wild-type mice. Levels of E. multilocularis-specific delayed-type hypersensitivity and serum antibodies were slightly decreased in LT alphaTNF-alpha -/- mice. This study shows that TNF-alpha and/or LT-alpha genes play an essential role in the immune protection mechanisms against E. multilocularis at the site of infection. PMID- 10476058 TI - Hormonal regulation of a cysteine proteinase gene, EPB-1, in barley aleurone layers: cis- and trans-acting elements involved in the co-ordinated gene expression regulated by gibberellins and abscisic acid. AB - The synthesis of EPB, a cysteine proteinase responsible for the degradation of seed endosperm storage proteins in barley (Hordeum vulgare), is induced by gibberellins (GA) and repressed by abscisic acid (ABA). The EPB gene family consists of two very similar members, EPB-1 and EPB-2, with the former being more highly induced by GA. We have functionally characterized the cis-acting elements in the EPB-1 promoter and determined that a gibberellin response element (GARE), a pyrimidine box and an upstream element are necessary for GA induction. By comparison with the promoters of alpha-amylase genes, which are also induced by GA, we suggest that GARE is coupled with the upstream element and the pyrimidine box to form a GA response complex. In addition, we have shown that the 3' untranslated/untranscribed region of the EPB-1 gene is required for a low background expression in the absence of GA. Constitutive expression of a transcription factor, GAMyb, in the absence of GA leads to the transactivation of EPB-1 expression in a dosage dependent manner with the highest level comparable to that in fully GA-induced tissue. Co-expression of a truncated version of GAMyb containing only the DNA binding domain blocks the GA-induction of EPB-1, further supporting the role of GAMyb in the regulation of gene expression. Although ABA is very effective in blocking the GA induction of EPB-1, it has no effect on the GAMyb-mediated expression of EPB-1. We suggest that ABA acts upstream of the formation of functional GAMyb which co-ordinates the hormonal regulation of a diverse group of genes in cereal aleurone layers, including those encoding EPB and alpha-amylases. PMID- 10476057 TI - Cloning and expression of a cDNA encoding an elongation factor 1beta/delta protein from Echinococcus granulosus with immunogenic activity. AB - A cDNA clone (Eg EF-1beta/delta) of Echinococcus granulosus has been isolated by an expression library screened with immunoglobulin (Ig)E of sera from patients with cystic echinococcosis (CE). The Eg EF-1beta/delta was identified on the basis of sequence homology to the subunits beta or delta of the elongation factor 1. The amino acid sequence deduced from this open reading frame is 244 residues long with a predicted molecular mass of 31 kDa. In Southern blot under high stringent condition, Eg EF-1beta/delta hybridized to genomic DNA of E.granulosus at two bands of 4 and 2.5 Kb. In immunoblotting analysis, the Eg EF-1beta/delta protein shows immunological reactivity with sera from CE patients: 51.7% of sera contained IgE, 41.7% IgG and 18.3% IgG4 specific to the recombinant protein. We identify the Eg EF-1beta/delta by immunoblotting with specific monoclonal antibody both in protoscoleces and in sheep hydatid fluid. The higher percentage of humoral immune response against Eg EF-1beta/delta observed in CE patients with calcified cysts than in patients with active cysts indicates the possible release of the protein in the hydatid fluid after protoscoleces degeneration suggesting the possible use of this antigen in the immunosurveillance of CE. Overall, these findings seem to assign to Eg EF-1beta/delta a key role in the allergic disorders and in the complex host-parasite relationship in CE. PMID- 10476059 TI - Decreased expression of sucrose phosphate synthase strongly inhibits the water stress-induced synthesis of sucrose in growing potato tubers. AB - Water stress stimulates sucrose synthesis and inhibits starch synthesis in wild type tubers. Antisense and co-suppression potato transformants with decreased expression of sucrose-phosphate synthase (SPS) have been used to analyse the importance of SPS for the regulation of this water-stress induced change in partitioning. (i) In the absence of water stress, a 70-80% decrease in SPS activity led to a 30-50% inhibition of sucrose synthesis and a slight (10-20%) increase of starch synthesis in tuber discs in short-term labelling experiments with low concentrations of labelled glucose. Similar changes were seen in short term labelling experiments with intact tubers attached to well-watered plants. Provided plants were grown with ample light and water, transformant tubers had a slightly lower water and sucrose content and a similar or even marginally higher starch content than wild-type tubers. (ii) When wild-type tuber slices were incubated with labelled glucose in the presence of mannitol to generate a moderate water deficit (between -0.12 and -0.72 MPa), there was a marked stimulation of sucrose synthesis and inhibition of starch synthesis. A similar stimulation was seen in labelling experiments with wild-type tubers that were attached to water-stressed wild-type plants. These changes were almost completely suppressed in transformants with a 70-80% reduction of SPS activity. (iii) Decreased irrigation led to an increase in the fraction of the dry-matter allocated to tubers in wild-type plants. This shift in allocation was prevented in transformants with reduced expression of SPS. (iv) The results show that operation of SPS and the sucrose cycle in growing potato tubers may lead to a marginal decrease in starch accumulation in non-stressed plants. However, SPS becomes a crucial factor in water-stressed plants because it is required for adaptive changes in tuber metabolism and whole plant allocation. PMID- 10476060 TI - Antisense inhibition of the GDP-mannose pyrophosphorylase reduces the ascorbate content in transgenic plants leading to developmental changes during senescence. AB - GDP-mannose pyrophosphorylase (GMPase, EC 2.7.7.22) catalyses the synthesis of GDP-D-mannose and represents the first committed step in the formation of all guanosin-containing sugar nucleotides found in plants which are precursors for cell wall biosynthesis and, probably more important, the synthesis of ascorbate. A full-length cDNA encoding GMPase from S. tuberosum was isolated. Transgenic potato plants were generated in which the GMPase cDNA was introduced in antisense orientation to the 35S promoter. Transformants with reduced GMPase activity were selected. Transgenic plants were indistinguishable from the wild-type when held under tissue culture conditions, however, a major change was seen 10 weeks after transfer into soil. Transgenic plants showed dark spots on leaf veins and stems with this phenotype developing from the bottom to the top of the plant. In case of the line with the strongest reduction, all aerial parts finally dried out after 3 months in soil, in contrast to the wild-type plants which did not start to senesce at this time. This coincides with a reduction of ascorbate contents in the transgenic plants, which is in agreement with the recently proposed pathway of ascorbate biosynthesis. Furthermore, leaf cell walls of the transgenic potato plants had mannose contents that were reduced to 30-50% of the wild-type levels, whereas the composition of tuber cell walls was unchanged. The glycosylation pattern of proteins was unaffected by GMPase inhibition, as studied by affinoblot analysis. PMID- 10476061 TI - AtAMT1 gene expression and NH4+ uptake in roots of Arabidopsis thaliana: evidence for regulation by root glutamine levels. AB - The mechanisms involved in regulating high-affinity ammonium (NH4+) uptake and the expression of the AtAMT1 gene encoding a putative high-affinity NH4+ transporter were investigated in the roots of Arabidopsis thaliana. Under conditions of steady-state nitrogen (N) supply, transcript levels of the AtAMT1 gene and Vmax values for high-affinity 13NH4+ influx were inversely correlated with levels of N provision. Following re-supply of NH4NO3 to N-starved plants, AtAMT1 mRNA levels and 13NH4+ influx declined rapidly but remained high when the conversion of NH4+ to glutamine (Gln) was blocked with methionine sulfoximine (MSX). This result demonstrates that end products of NH4+ assimilation, rather than NH4+ itself, are responsible for regulating AtAMT1 gene expression. Consistent with this hypothesis, AtAMT1 gene expression and NH4+ influx were suppressed by provision of Gln alone, or together with NH4NO3 plus MSX. Furthermore, AtAMT1 transcript levels and 13NH4+ influx were negatively correlated with root Gln concentrations, following re-supply of N to N-starved plants. In addition to this level of control, the data suggest that high cytoplasmic [NH4+] may inhibit NH4+ influx. PMID- 10476062 TI - Suppression of pea nuclear topoisomerase I enzyme activity by pea PCNA. AB - Proliferating cell nuclear antigen (PCNA), a highly conserved DNA polymerase accessory protein of eukary- otic kingdom, has not been studied thoroughly in bio chemical terms in plants. We describe the isolation of the cDNA encoding PCNA from the pea cDNA library using the PCR approach. The cDNA was used for expression of pea PCNA in bacteria as a fusion protein (GST.PCNA) with the GST tag at the amino terminal end. The GST.PCNA stimulated the partially purified pea DNA polymerases approximately 30-fold. The stimulation was due to the oligomeric form of GST.PCNA. The pea PCNA interacted with the recombinant type I pea topoiso merase as well as the native pea nuclear topoisomerase I and repressed the DNA relaxation activities. However, the DNA binding activity of Topo I remained undisturbed in the presence of high amounts of PCNA, thereby signify- ing that the catalysis of Topo I was probably affected by PCNA. PMID- 10476063 TI - Deficiency in phytoalexin production causes enhanced susceptibility of Arabidopsis thaliana to the fungus Alternaria brassicicola. AB - The phytoalexin-deficient Arabidopsis mutant pad3-1, which is affected in the production of the indole-type phytoalexin camalexin, has previously been shown not to display altered susceptibility to either the bacterium Pseudomonas syringae (Glazebrook & Ausubel 1994; Proc. Natl. Acad. Sci. USA, 91: 8955-8959) or the biotrophic fungi Peronospora parasitica (Glazebrook et al. 1997; Genetics, 146: 381-392) and Erysiphe orontii (Reuber et al. 1998; Plant J. 16: 473-485). We now show that this mutant is markedly more susceptible than its wild-type parental line to infection by the necrotrophic fungus Alternaria brassicicola, but not to Botrytis cinerea. A strong camalexin response was elicited in wild type plants inoculated with either Alternaria brassicicola or Botrytis cinerea, whereas no camalexin could be detected in pad3-1 challenged with these fungi. Hence, PAD3 appears to be a key determinant in resistance to at least A. brassicicola. The induction of salicylate-dependent and jasmonate/ethylene dependent defense genes was not reduced in Alternaria-challenged pad3-1 plants compared to similarly treated wild-type plants. Camalexin production could not be triggered by exogenous application of either salicylate, ethylene or jasmonate and was not, or not strongly, reduced in mutants with defects in perception of these defense-related signal molecules. Camalexin-production appears to be controlled by a pathway that exhibits little cross-talk with salicylate-, ethylene- and jasmonate-dependent signalling events. PMID- 10476064 TI - Transcription activation mediated by the bZIP factor SPA on the endosperm box is modulated by ESBF-1 in vitro. AB - A modified in vitro transcription system has been used to study the function of the cloned bZIP transcription factor SPA and the binding activity ESBF I in activating transcription from the bifactorial endosperm box region of the wheat prolamin LMWG-1D1 gene. Recombinant SPA expressed in Escherichia coli activated transcription from the endosperm box motif, and this was dependent upon the binding of the nuclear protein ESBF I. ESBF I did not activate transcription independently, but potentiated SPA-mediated transcriptional activation. ESBF I is likely to be the equivalent of, or contain the recently characterised DOF class of, Zn-finger protein called WPBF. These data provide new information about the interplay of members of the bZIP and DOF transcription factor families in regulating expression from bifactorial sites found in a variety of plant promoters. PMID- 10476065 TI - Flavonoid hydroxylase from Catharanthus roseus: cDNA, heterologous expression, enzyme properties and cell-type specific expression in plants. AB - We investigated the P450 dependent flavonoid hydroxylase from the ornamental plant Catharanthus roseus. cDNAs were obtained by heterologous screening with the CYP75 Hf1 cDNA from Petunia hybrida. The C. roseus protein shared 68-78% identity with other CYP75s, and genomic blots suggested one or two genes. The protein was expressed in Escherichia coli as translational fusion with the P450 reductase from C. roseus. Enzyme assays showed that it was a flavonoid 3', 5'-hydroxylase, but 3'-hydroxylated products were also detected. The substrate specificity was investigated with the C. roseus enzyme and a fusion protein of the Petunia hybrida CYP75 with the C. roseus P450 reductase. Both enzymes accepted flavanones as well as flavones, dihydroflavonols and flavonols, and both performed 3'- as well as 3'5'-hydroxylation. Kinetics with C. roseus cultures on the level of enzyme activity, protein and RNA showed that the F3'5'H was present in dark-grown cells and was induced by irradiation. The same results were obtained for cinnamic acid 4-hydroxylase and flavanone 3beta-hydroxylase. In contrast, CHS expression was strictly dependent on light, although CHS is necessary in the synthesis of the F3'5'H substrates. Immunohistochemical localization of F3'5'H had not been performed before. A comparison of CHS and F3'5'H in cotyledons and flower buds from C. roseus identified CHS expression preferentially in the epidermis, while F3'5'H was only detected in the phloem. The cell-type specific expression suggests that intercellular transport may play an important role in the compartmentation of the pathways to the different flavonoids. PMID- 10476066 TI - Cloning and expression of a cDNA encoding homospermidine synthase from Senecio vulgaris (Asteraceae) in Escherichia coli. AB - The enzyme homospermidine synthase catalyzes the NAD+-dependent conversion of 2 mol putrescine into homospermidine. Instead of putrescine, spermidine can substitute for the first putrescine moiety in plants, in which case diaminopropane instead of ammonia is released. The enzyme facilitates the formation of the 'uncommon' polyamine homospermidine which is an important precursor in the biosynthesis of pyrrolizidine alkaloids. The first plant homospermidine synthase was purified to apparent chemical homogenity from the root tissue culture Senecio vernalis (Asteraceae) (Bottcher et al. 1994, Can. J. Chem. 72, 80-85; Ober 1997, Dissertation). Four endopeptidase LysC fragments were sequenced from the purified protein. With the aid of degenerate primers against these peptides, a cDNA encoding homospermidine synthase was now cloned and characterized from Senecio vulgaris. The nucleotide sequence of the cloned cDNA revealed an open reading frame of 1155-base pairs containing 385 amino acids with a predicted Mr of 44500. GenBank research revealed that the deduced amino acid sequence shows 59% identity to human deoxyhypusine synthase. The homospermidine synthase encoding cDNA was subcloned into the expression vector pet15b and overexpressed in E. coli. The recombinant enzyme formed upon expression catalyzed homospermidine synthesis. PMID- 10476067 TI - Short Communication: An apospory-specific genomic region is conserved between Buffelgrass (Cenchrus ciliaris L.) and Pennisetum squamulatum Fresen. AB - Twelve molecular markers linked to pseudogamous apospory, a form of gametophytic apomixis, were previously isolated from Pennisetum squamulatum Fresen. No recombination between these markers was found in a segregating population of 397 individuals (Ozias-Akins et al. 1998, Proc. Natl Acad. Sci. USA, 95, 5127-5132). The objective of the present study was to test if these markers were also linked to the aposporous mode of reproduction in two small segregating populations of Cenchrus ciliaris (= Pennisetum ciliare (L.)Link), another apomictic grass species. Among 12 markers (sequence characterized amplified regions, SCARs), six were scored as dominant markers between aposporous and sexual C. ciliaris genotypes (presence/absence, respectively). Five were always linked to apospory and one showed a low level of recombination in 84 progenies. Restriction fragment length polymorphisms (RFLPs) were observed between sexual and apomictic phenotypes for three of the six remaining SCARs from P. squamulatum when used as probes. No recombination was observed in the F1 progenies. Preliminary data from megabase DNA analysis and sequencing in both species indicate that an apospory specific genomic region (ASGR) is highly conserved between the two species. Although C. ciliaris has a smaller genome size to P. squamulatum, a higher copy number for markers linked to apospory found in the former may impair the progress of positional cloning of gene(s) for apomixis in this species. PMID- 10476068 TI - Technical Advance: Stable chloroplast transformation in potato: use of green fluorescent protein as a plastid marker. AB - We describe here the development of a reproducible plastid transformation system for potato and regeneration of plants with uniformly transformed plastids. Two distinct tobacco-specific plastid vectors, pZS197 (Prrn/aadA/TpsbA) and pMON30125 (Prrn/GFP/Trps16:PpsbA/aadA/TpsbA), designed for integration into the large single copy and inverted repeat regions of the plastid genome, respectively, were bombarded into leaf explants of potato line FL1607. A total of three transgenic lines were selected out of 46 plates bombarded with pZS197 and three transgenic lines out of 104 plates were obtained with pMON30125. Development of a high frequency leaf-based regenera- tion system, a stringent selection scheme and optimization of biolistic transformation protocol were critical for recovery of plastid transformants. Plastid-expressed green fluorescent protein was used as a visual marker for identification of plastid transformants at the early stage of selection and shoot regeneration. The establishment of a plastid transformation system in potato, which has several advantages over routinely used nuclear transformation, offers new possibilities for genetic improvement of this crop. PMID- 10476069 TI - Technical Advance: Differential extraction of hydrophobic proteins from chloroplast envelope membranes: a subcellular-specific proteomic approach to identify rare intrinsic membrane proteins. AB - Identification of rare hydrophobic membrane proteins is a major biological problem that is limited by the specific biochemical approaches required to extract these proteins from membranes and purify them. This is especially true for membranes, such as plastid envelope membranes, that have a high lipid content, present a wide variety of specific functions and therefore contain a large number of unique, but minor, proteins. We have optimized a procedure, based on the differential solubilization of membrane proteins in chloroform/methanol mixtures, to extract and concentrate the most hydrophobic proteins from chloroplast envelope membrane preparations, while more hydrophilic proteins were excluded. In addition to previously characterized chloroplast envelope proteins, such as the phosphate/triose phosphate translocator, we have identified new proteins that were shown to contain putative transmembrane alpha-helices. Moreover, using different chloroform/methanol mixtures, we have obtained differential solubilization of envelope proteins as a function of their hydrophobicity. All the proteins identified were genuine chloroplast envelope proteins, most of them being localized within the inner membrane. Our procedure enables direct mapping (by classical SDS-PAGE) and identification of hydrophobic membrane proteins, whatever their isoelectric point was, that are minor components of specific subcellular compartments. Thus, it complements other techniques that give access to peripheral membrane proteins. If applied to various cell membranes, it is anticipated that it can expedite the identification of hydrophobic proteins involved in transport systems for ions or organic solutes, or it may act as signal receptors or to control metabolic processes and vesicle trafficking. PMID- 10476070 TI - Technical Advance: Confocal measurement of the three-dimensional size and shape of plant parenchyma cells in a developing fruit tissue. AB - Parenchyma cells from the inner mesocarp of a grape berry (Vitis vinifera L. cv. Chardonnay) were visualised in three-dimensions within a whole mount of cleared, stained tissue using confocal laser scanning microscopy and digital image reconstruction. The whole berry was fixed, bisected longitudinally, cleared in methyl salicylate, stained with safranin O and mounted in methyl salicylate. Optical slices were collected at 1.0 um intervals to a depth of 150 um. Neighbouring z-series were joined post-collection to double the field-of-view. Attenuation at depth of the fluorescent signal from cell walls was quantified and corrected. Axial distortion due to refractive index mismatch between the immersion and mounting media was calibrated using yellow-green fluorescent microspheres and corrected. Transmission electron microscopy was used to correct fluorescent measurements of cell wall thickness. Digital image reconstructions of wall-enclosed spaces enabled cells to be rendered as geometric solids of measurable surface area and volume. Cell volumes within the inner mesocarp tissue of a single grape berry exhibited a 14-fold range, with polysigmoidal distribution and groupings around specific size classes. Cell shape was irregular and the planes of contact were rarely flat or simple. Variability in cell shape was indicated by the range in surface area to volume ratios, from 0.080 to 0.198 um-1. Structural detail at the internal surface of the cell wall was apparent. The technique is applicable to a wide range of morphometric analyses in plant cell biology, particularly developmental studies, and reveals details of cell size and shape that were previously unattainable. PMID- 10476071 TI - Cloning and characterisation of PGA1 and PGA2: two G protein alpha-subunits from pea that promote growth in the yeast Saccharomyces cerevisiae. AB - We report here on the cloning and characterization of two G protein alpha subunits from pea: PGA1 and PGA2. Based on DNA gel blot analysis, PGA1 and PGA2 are the only Galpha homologous sequences in pea. RT-PCR analysis reveals that PGA1 and PGA2 transcripts are present in a variety of adult pea tissues. However, PGA2 mRNA is consistently detected at a lower level than PGA1 and demonstrates some degree of tissue specificity relative to PGA1. In the apical bud of pea seedlings, PGA1 and PGA2 transcripts decrease in response to 24 h of white light following growth for 6 days in darkness. The G protein mediated, yeast mating pathway was used to analyse the function of PGA1 and PGA2 in vivo. PGA1 downregulates the mating pathway, but through a mechanism that is independent of Gbetagamma sequestration. Unexpectedly, both PGA1 and PGA2 promote growth through a mating pathway independent mechanism. PMID- 10476072 TI - Arabidopsis ovule is the target for Agrobacterium in planta vacuum infiltration transformation. AB - The visual marker GUS has been utilized in this study to understand the Arabidopsis thaliana vacuum infiltration transformation process by Agrobacterium tumefaciens. High transformation frequencies of up to 394 transgenic seeds per infiltrated plant were achieved. The results showed that the majority of the transgenic seeds from single infiltrated plants were from independent transformation events based on Southern analysis, progeny segregation, distribution of transgenic seeds throughout the infiltrated plants and the microscopic analysis of GUS expression in ovules of infiltrated plants. GUS expression in mature pollen and anthers was monitored daily from 0 to 12 days post-infiltration. In addition, all ovules from a single infiltrated plant were examined every other day. GUS expression frequencies of up to 1% of pollen were observed 3-5 days post-infiltration, whereas frequencies of up to 6% were detected with ovules of unopened flowers 5-11 days post-infiltration. Most importantly, transgenic seeds were obtained only from genetic crosses using infiltrated plants as the pollen recipient but not the pollen donor, demonstrating Agrobacterium transformation through the ovule pathway. PMID- 10476073 TI - CYP78A5 encodes a cytochrome P450 that marks the shoot apical meristem boundary in Arabidopsis. AB - The normal development of shoot structures depends on controlling the growth, proliferation and differentiation of cells derived from the shoot apical meristem. We have identified the CYP78A5 gene encoding a putative cytochrome P450 monooxygenase that is the first member of the CYP78 family from Arabidopsis. This gene is strongly expressed in the peripheral regions of the vegetative and reproductive shoot apical meristems, defining a boundary between the central meristematic zone and the developing organ primordia. In addition, CYP78A5 shows a dynamic pattern of expression during floral development. Overexpression of CYP78A5 affects multiple cell types, causing twisting and kinking of the stem and defects in floral development. To define the relationship of CYP78A5 to genes controlling meristem function, we examined CYP78A5 expression in plants mutant for SHOOT MERISTEMLESS, ZWILLE and ARGONAUTE, and have found that CYP78A5 expression is altered in these mutant backgrounds. We propose that CYP78A5 has a role in regulating directional growth in the peripheral region of the shoot apical meristem in response to cues established by genes regulating meristem function. PMID- 10476074 TI - The AtSUC1 sucrose carrier may represent the osmotic driving force for anther dehiscence and pollen tube growth in Arabidopsis. AB - The Arabidopsis AtSUC1 protein has previously been characterized as a plasma membrane H+-sucrose symporter. This paper describes the sites of AtSUC1 gene expression and AtSUC1 protein localization and assigns specific functions to this sucrose transporter in anther development and pollen tube growth. RNase protection assays revealed AtSUC1 expression exclusively in floral tissue, which was confirmed by analyses of AtSUC1 promoter-beta-glucuronidase (GUS) plants. In situ hybridizations identified AtSUC1 expression in anther connective tissue, in funiculi and in fully developed pollen grains. Indirect immuno-fluorescence analyses with anti-AtSUC1 antiserum confirmed AtSUC1 protein localization in the connective tissue and funiculi. In mature pollen grains, however, despite high AtSUC1 mRNA levels no AtSUC1 protein was found. Only after pollination of stylar papillae was AtSUC1 protein detected inside the pollen and later inside the growing pollen tubes, suggesting a translation of pre-existing AtSUC1 mRNA after pollination. Pollen germination analyses underlined the important role of sucrose for pollen tube growth. The data presented suggest a role of AtSUC1 in the controlled dehiscence of Arabidopsis anthers. It is postulated that an important function of AtSUC1 is the cell-specific modulation of water potentials. PMID- 10476075 TI - The Arabidopsis blue light receptor cryptochrome 2 is a nuclear protein regulated by a blue light-dependent post-transcriptional mechanism. AB - Cryptochrome 2 is a flavin-type blue light receptor mediating floral induction in response to photoperiod and a blue light-induced hypocotyl growth inhibition. cry2 is required for the elevated expression of the flowering-time gene CO in response to long-day photoperiods, but the molecular mechanism underlying the function of cry2 is not clear. The carboxyl domain of cry2 bears a basic bipartite nuclear localization signal, and the cry2 protein was co-fractionated with the nucleus. Analysis of transgenic plants expressing a fusion protein of CRY2 and the reporter enzyme GUS (GUS-CRY2) indicated that the GUS-CRY2 fusion protein accumulated in the nucleus of transgenic plants grown in dark or light. The C-terminal domain of cry2 that contains the basic bipartite nuclear localization signal was sufficient to confer nuclear localization of the fusion protein. Phenotypic analysis of transgenic plants expressing the fusion protein GUS-CRY2 demonstrated that GUS-CRY2 acts as a functional photoreceptor in vivo, mediating the blue light-induced inhibition of hypocotyl elongation. These results strongly suggest that cry2 is a nuclear protein. Although no obvious light regulation was found for the nuclear compartmentation of GUS-CRY2 fusion protein, the abundance of GUS-CRY2 was regulated by blue light in a way similar to that of cry2. PMID- 10476076 TI - Nuclear localization of the Arabidopsis blue light receptor cryptochrome 2. AB - The cryptochrome blue light photoreceptor family of Arabidopsis thaliana consists of two members, CRY1 and CRY2 (PHH1). CRY2 contains a putative nuclear localization signal (NLS) within its C-terminal region. We examined whether CRY2 is localized in the nucleus and whether the C-terminal region of CRY2 is involved in nuclear targeting. Total cellular and nuclear protein extracts from Arabidopsis were subjected to immunoblot analysis with CRY2-specific antibodies. Strong CRY2 signals were obtained in the nuclear fraction. Fusion proteins consisting of the green fluorescent protein (GFP) and different fragments of CRY2 were expressed in parsley protoplasts and the localization of the fusion proteins was determined by fluorescence and confocal laser scanning microscopy. GFP fusions containing the entire CRY2 protein or its C-terminal region were found exclusively in the nucleus. We conclude from these results that CRY2 is localized in the nucleus and that nuclear localization is mediated by the C-terminal region of CRY2. PMID- 10476077 TI - MAP kinase activation by hypoosmotic stress of tobacco cell suspensions: towards the oxidative burst response? AB - Hypoosmotic stress activates a phosphorylation-dependent oxidative burst. In-gel kinase assays were performed to characterize the protein kinases that could be implicated in osmoregulation and in the activation of the oxidative burst. Hypoosmotic stress activated several kinases among which 50 and 46 kDa proteins displayed mitogen-activated protein kinase (MAP kinase) properties. They phosphorylated myelin basic protein in the absence of calcium, were recognized by antibodies directed against human MAP kinases, and were phosphorylated on tyrosine. Immunoprecipitation with an antibody directed against the tobacco MAP kinase Ntf4 showed that at least one of the activated kinases would be Ntf4-like. Apigenin, a MAP kinase and cyclin-dependent kinase inhibitor which prevents the hypoosmotically induced oxidative burst (Cazale et al. 1998; Plant Physiol. 116, 659-669), inhibited these kinases in vitro suggesting that they may play a role in the activation of the oxidative burst. Like the oxidative response, activation of the kinases depended on extracellular calcium influx and protein kinases sensitive to staurosporine and 6-DMAP. However, kinase activation did not depend on effluxes through anion channels or on the oxidative burst. Two-dimensional in gel kinase assays revealed the presence of three protein kinases with an apparent molecular mass of 50 kDa and one of 46 kDa, all four being activated by hypoosmotic stress. The same kinases were also activated by oligogalacturonides and salicylic acid, underlying the importance of these MAP kinases as common components of different signaling pathways triggered by different extracellular stimuli. PMID- 10476079 TI - Yariv reagent treatment induces programmed cell death in Arabidopsis cell cultures and implicates arabinogalactan protein involvement. AB - Arabinogalactan proteins (AGPs) are a family of highly glycosylated, hydroxyproline-rich glycoproteins implicated in various aspects of plant growth and development. (beta-D-glucosyl)3 and (beta-D-galactosyl)3 Yariv phenylglycosides, commonly known as Yariv reagents, specifically bind AGPs in a non-covalent manner. Here (beta-D-galactosyl)3 Yariv reagent was added to Arabidopsis thaliana cell suspension cultures and determined to induce programmed cell death (PCD) by three criteria: (i) DNA fragmentation as detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) of DNA 3'-OH groups; (ii) inter- nucleosomal DNA fragmentation as visualized by genomic Southern blotting; and (iii) structural changes characteristic of PCD including cytoplasmic shrinkage and condensation, chromatin condensation and nuclear membrane blebbing. These findings implicate AGP involvement in PCD in plants, presumably by perturbation of AGPs located at the plasma membrane-cell wall interface. PMID- 10476078 TI - Dimerization and DNA binding of auxin response factors. AB - Auxin response factors (ARFs) are transcription factors that bind with specificity to TGTCTC auxin response elements (AuxREs) found in promoters of primary/early auxin response genes. ARFs are encoded by a multi-gene family, consisting of more than 10 genes. Ten ARFs have been analyzed by Northern analysis and were found to be expressed in all major plant organs and suspension culture cells of Arabidopsis. The predicted amino acid sequences indicate that the 10 ARFs contain a novel amino-terminal DNA binding domain and a carboxyl terminal dimerization domain, with the exception of ARF3 which lacks this dimerization domain. All ARFs tested bind with specificity to the TGTCTC AuxRE, but there are subtle variations in the sequence requirements at positions 5 (T) and 6 (C) of the AuxRE. While the amino-terminal domain of about 350 amino acids is sufficient for binding ARF1 to TGTCTC AuxREs, this domain is not sufficient for the binding of some other ARFs to palindromic AuxREs. Our results suggest that ARFs must form dimers on palindromic TGTCTC AuxREs to bind stably, and this dimerization may be facilitated by conserved motifs found in ARF carboxyl terminal domains. Dimerization in at least some cases may dictate which ARF(s) are targeted to AuxREs. PMID- 10476080 TI - In vivo analysis of plastid psbA, rbcL and rpl32 UTR elements by chloroplast transformation: tobacco plastid gene expression is controlled by modulation of transcript levels and translation efficiency AB - 5' and 3' untranslated regions (UTRs) of plastid RNAs act as regulatory elements for post-transcriptional control of gene expression. Polyethylene glycol-mediated plastid transformation with UTR-GUS reporter gene fusions was used to study the function of the psbA, rbcL and rpl32 UTRs in vivo. All gene fusions were expressed from the same promoter, i.e. the promoter of the 16S-rRNA gene, such that variations in RNA and protein levels would be due to the involved UTR elements alone. Transgenic tobacco lines containing different combinations of UTRs showed fivefold variation in the uidA-mRNA level (RNA stability) and approximately 100-fold differences in GUS activity, a measure of translation activity. The rbcL 5'-UTR conferred greater mRNA stability than the psbA 5'-UTR on uidA transcripts. In contrast, the psbA 5'-UTR enhanced translation of GUS to a much greater extent compared to the rbcL 5'-UTR. The psbA 5'-UTR also mediated light-induced activation of translation which was not observed with other constructs. Deletion mutagenesis of an unanalysed terminal sequence element of the psbA 5'-UTR resulted in a twofold drop in uidA-mRNA level and a fourfold decrease in translation efficiency. Exchange of 3'-UTRs results in up to fivefold changes of mRNA levels and does not significantly influence translation efficiency. The mechanical impacts of these results on plastid translation regulation are discussed. PMID- 10476081 TI - Rhizobium nod factors induce increases in intracellular free calcium and extracellular calcium influxes in bean root hairs AB - Application of Nod factors to growing, responsive root hairs of the bean Phaseolus vulgaris induces marked changes in both the intracellular cytosolic free calcium (Ca2+) and in the influx of extracellular [Ca2+]. The intracellular [Ca2+], which has been measured by ratiometric imaging in cells microinjected with fura-2-dextran (70 kDa), elevates within 5 min from approximately 400 nM to 1500 nM in localised zones in the root hair apex. Of particular note is the observation that the elevated regions of [Ca2+] appear to shift position during short time intervals. Increases in and fluctuations of the intracellular [Ca2+] are also observed in the perinuclear region after 10-15 min treatment with Nod factors. The extracellular Ca2+ flux, detected with the non-invasive, calcium specific vibrating electrode, is inwardly directed and also increases quickly in response to Nod factors from 13 pmol cm-2 s-1 to 28 pmol cm-2 s-1. Chitin oligomers, which are structurally similar but biologically inactive when compared to the active Nod factors, fail to elicit changes in either intracellular or extracellular Ca2+. The similar timing and location of the intracellular elevations and the increased extracellular influx provide support for the idea that Ca2+ participates in secretion and cell wall remodelling, which occur in anticipation of root hair deformation and curling. PMID- 10476082 TI - A synthetic gene coding for the green fluorescent protein (GFP) is a versatile reporter in Chlamydomonas reinhardtii. AB - The use of Chlamydomonas reinhardtii as a model system has been hindered by difficulties encountered in expressing foreign genes. We have synthesised a gene encoding green fluorescent protein (GFP) adapted to the codon usage of C. reinhardtii (cgfp). After verifying the gene was functional in Escherichia coli, the cgfp was fused in frame to the phleomycin resistance gene ble from Streptoalloteichus hindustanus and expressed in C. reinhardtii under control of the rbcS2 promoter and intron sequences. The GFP-fluorescence was seen only in the nucleus demonstrating the nuclear accumulation of the Ble-GFP fusion protein. The cgfp was also fused to the chlamyopsin gene, cop, and expressed in C. reinhardtii under control of the cop promoter. The eyespot became fluorescent indicating that the opsin-GFP fusion protein was correctly directed into the eyespot along with the endogenous unmodified opsin. We conclude that cgfp provides a useful tool to visualize protein synthesis and localisation in vivo in C. reinhardtii and possibly in related green algal species. PMID- 10476083 TI - Accurate and high resolution in situ hybridization analysis of gene expression in secondary stem tissues. AB - Accurate in situ hybridization analysis in secondary stem tissues of plants has been hindered by specific characteristics of these tissues. First, secondary cell walls non-specifically bind probes used for in situ hybridization thus preventing gene expression analysis in the lignified regions of the stem, such as the xylem. Second, the mRNA in the cambial meristem and its recent derivatives are prone to inadequate fixation when conventional techniques are used. Here we describe an in situ hybridization technique which uses fast freezing and freeze substitution to cryoimmobilize the mRNA followed by embedding in a methacrylate resin for high resolution analysis of gene expression. By using a transgenic poplar line harbouring rolC:uidA, rolC:iaaM, the gene expression pattern could be compared with histochemical GUS staining. This in situ hybridization technique results in superior preservation of cellular contents, retention of mRNA in all cell types in the poplar stem, a significant reduction of non-specific binding to secondary cell walls and a resolution not previously possible in secondary tissues. This technique will be particularly valuable for the expression analysis of genes involved in xylogenesis and wood formation. PMID- 10476084 TI - Severe neutropenia as an adverse effect of methimazole in the treatment of hyperthyroidism. AB - Ms. K., a white, 47-year-old female with a history of hyperthyroidism had been treated with methimazole daily for a period of 9 years. She presented with a 2 day history of fever higher than 103 degrees F and cellulitis of the right arm after a scratch injury. White blood cell count (WBC) was noted at 0.4 x 10(3)/microL and neutrophils at 5.6%, indicating agranulocytosis. Methimazole was discontinued by the patient with the onset of symptoms. Appropriate intravenous antibiotic therapy and reverse isolation were provided in the acute-care setting, as well as administration of the granulocyte colony-stimulating factor (G-CSF) filgrastim. No recovery of the granulocyte count or improvement of clinical condition was noted until her sixth day of admission, at which time her WBC increased to 2.6 x 10(3)/microL. The administration of intravenous antifungals and antibiotics prevented overwhelming sepsis, while giving the G-CSF the opportunity to stimulate growth of granulocytes to finally fight the offending organisms and save this patient. PMID- 10476085 TI - Community-acquired pneumonia in the elderly. PMID- 10476086 TI - Culturalizing health care for a culturally diverse population: the Amish. AB - Advanced practice nurses (APNs) are often the major source of primary care for culturally diverse groups. Working with patients who have values that differ from the mainstream culture can make delivery of health care complex and challenging. The purpose of this qualitative study was to use grounded theory methods to explore the process by which APNs develop and maintain therapeutic relationships with a culturally unique group, the Amish. Nine APNs who practiced primarily with Amish patients were interviewed about their strategies to promote therapeutic relationships. The process of culturalizing health care emerged as the strategy used by the APNs. Specific themes were learning the culture, developing a relationship, individualizing care, being comfortable with abilities, and working two systems. Rather than expecting patients to conform to the mainstream cultural beliefs and practices, APN informants used a variety of methods that allowed Amish patients to blend self-care practices with conventional (i.e., English care). PMID- 10476088 TI - The San Antonio Biethnic Children's Blood Pressure Study: anthropometric findings. AB - Cardiovascular diseases and diabetes are very prevalent today, and early detection of the precursors of these diseases is important. Increased weight and body mass indexes (BMIs) are known to be risk factors for these adult diseases. The objectives of this portion of the study were to obtain anthropometric measurements known to correlate with children's blood pressure levels and to compare these measurements among Mexican-American (M-A) females and males and non Hispanic white (W) females and males. These data will allow healthcare providers to know normal weights and BMIs for different ethnic groups so that early detection of risk factors can be done. Anthropometric measurements (weight, height, BMI, subscapular and triceps skinfold thickness, and arm circumference) were obtained on 4,195 M-A and 2,039 W children in kindergarten through 12th grade in two school districts in the San Antonio, Texas, area. Data were statistically analyzed and related to age. M-A males had a tendency to be heavier than W males throughout the ages studied. M-A females had a tendency to be heavier than W females until 11 years of age, but that trend was reversed between ages 14 and 18. For all four subgroups, there was a marked skewness toward obesity based on BMI (kg/m2). Height of M-A children beginning as early as 5 years of age was slightly but significantly (P > or = .05) less than that of W children of the same gender. M-A males and females had either a tendency for or significantly higher values (P > or = .05) for BMI than W counterparts. M-A children had larger subscapular and triceps skinfold thickness than their W counterparts, with females showing greater values than males. The data indicate that there are several important differences in weight, height, BMI, and skinfold thickness between M-A and W children and between boys and girls as noted above. There are also considerable increases in weight with a relatively stable height in all four subgroups since the last published growth charts. PMID- 10476087 TI - A nursing screening program for diabetes in northern New Mexico. AB - This paper presents the results of a community screening program conducted by nurses in northern New Mexico. Seven hundred and ninety-nine persons were screened for diabetes following American Diabetes Association guidelines. Screening procedures included having participants complete a questionnaire and testing them for capillary blood glucose levels. Initial results found a 5.7% positive screening rate. A large proportion of those who tested positive (80%) were unavailable for follow-up procedures. PMID- 10476089 TI - Nursing scholarship: role of faculty practice. AB - As nursing education has moved into the academic setting, nursing faculty have focused on the traditional components of scholarship in the faculty role: research and publication, teaching, and service. However, dramatic changes have occurred in higher education and health care forcing nursing faculty to reexamine their roles in both systems. A survey was conducted to explore the views held by deans and faculty of colleges of nursing on scholarship and to describe ways in which these views on scholarship reflect faculty practice. Principal findings of this study indicated that nursing deans and faculty view scholarship as the generation, dissemination, application, and advancement of nursing knowledge. The components of scholarship in the nursing faculty role were described as research and publication, teaching, service, faculty practice, and presentations. The strongest theme to emerge from the study was the role conflict and fragmentation experienced by faculty as they tried to balance all the components of scholarship. The topic of faculty practice was considered to be an important issue in nursing, with the majority of nursing faculty involved in faculty practice in environments where it is not accommodated, mandated, or formally expected. The findings of this study suggest that faculty practice roles could be considered a component of scholarship as long as scholarly outcomes are demonstrated. PMID- 10476090 TI - Ask for a nurse practitioner. PMID- 10476091 TI - Nursing town and nursing gown: time, space, and the reinvention of nursing through collaboration. AB - This paper argues that nursing can reinvent itself through collaboration between the clinical domain and the university and that this reinvention requires new ways of thinking about the relationship between town and gown. The paper analyzes the metaphor of the town/gown distinction and the constitution of time and space and their changing interrelationship. It offers a description of the learning environment of collaboration, which identifies some of the challenges that face nursing in each area. It then presents a way of envisioning the reinvention of nursing in this context, which will facilitate learning. PMID- 10476092 TI - A professional curriculum vitae will open career doors. AB - In today's challenging healthcare environment, it is essential for nurse practitioners to be able to describe themselves professionally on paper to compete for practice and academic opportunities. Nurse practitioners are competing with physician assistants as well as physicians for primary and acute care positions. A carefully compiled curriculum vitae will present the individual in the best light possible to help open career doors and enhance chances of success. Preparing a curriculum vitae will serve to highlight relevant professional accomplishments, whatever the setting, toward the fulfillment of professional goals. This article reviews the current professional print and electronic literature on preparing a curriculum vitae to assist the nurse practitioner in developing this vital document. PMID- 10476093 TI - Taking charge of your computer: bookmarks and cookies. PMID- 10476094 TI - Profiles of editorial board members: New Zealand to San Diego. PMID- 10476095 TI - Get 'stroke smart'. PMID- 10476096 TI - Surviving billing scrutiny. PMID- 10476097 TI - OTC treatment for candidiasis. PMID- 10476098 TI - Getting it right the first time. PMID- 10476099 TI - An expanding landscape. Osteoporosis. Treatment options today. AB - Choices for osteoporosis therapy have expanded within the past 5 years. This article provides an overview of currently available therapy options. Exogenous estrogen can prevent and treat osteoporosis and is available in several delivery routes. Calcitonin is also designed to reduce bone loss in osteoporosis. Bisphosphonates such as alendronate prevent bone resorption by inhibiting osteoclasts and causing increased osteoclast cell death. Raloxifene is a selective estrogen receptor modulator and is the newest osteoporosis medication on the market. It may also have beneficial effects on breast cancer risk. All postmenopausal women should obtain 1,000 mg to 1,500 mg of calcium and 400 IU to 800 IU of vitamin D every day--regardless of any prescription therapy regimen for osteoporosis. They should also perform weight-bearing exercise, such as walking, for 20 to 30 minutes every day or for 1 hour three times a week. PMID- 10476100 TI - Diabetes in the older adult. Special considerations for age-related pathology. PMID- 10476101 TI - A behavioral approach to Alzheimer's disease. The progressively lowered stress threshold model. PMID- 10476102 TI - Pharyngitis in children. Determining the etiology is essential. PMID- 10476103 TI - Parkinson's disease. Pieces of the diagnostic puzzle. PMID- 10476104 TI - Practical issues in asthma management. Self-care plans for adult patients. PMID- 10476105 TI - Chronic venous insufficiency. Breaking the cycle of ulceration. PMID- 10476106 TI - Candidiasis in children with HIV. A protocol for diagnosis and treatment. PMID- 10476107 TI - Yes Virginia, there is a glut. PMID- 10476108 TI - Life beyond the year 2000. PMID- 10476109 TI - The physical and psycho-social experiences of patients attending an outpatient medical oncology department: a cross-sectional study. AB - The aim of this study was to assess the prevalence and predictors of physical symptoms, anxiety, depression and perceived needs among patients receiving treatment at an outpatient medical oncology department using a cross-sectional survey. It was carried out at the outpatient clinic of an academic medical oncology department, which sees around 150-180 outpatients each week; 201 patients were selected. These patients answered questions to assess their levels of anxiety and depression (Hospital Anxiety and Depression Scale), perceived needs (Cancer Needs Questionnaire) and the frequency and severity of 15 physical symptoms. Fatigue, nausea, appetite loss and vomiting were the most commonly experienced and most debilitating physical symptoms. Approximately 25% of participants had borderline or clinical levels of anxiety and depression. Although relatively low levels of perceived needs were reported, physical and psychological needs were the most common. Levels of each outcome measure tended to be predictive of each other. Medical oncology outpatients experience a wide range of physical and psycho-social problems which appear, to some extent, interrelated. PMID- 10476110 TI - Patients' preferences for video cassette recorded information: effect of age, sex and ethnic group. AB - The emotional turmoil patients endure following a diagnosis of cancer can impair their ability to retain complex treatment-related information. Manoeuvres which increase the intensity of information have been shown to increase the amount retained. Providing details of treatment in a video format is one method of intensifying information provision, but the attitudes of patients to this format have not previously been evaluated. In this pilot study, the attitudes of 300 patients to video directed information were evaluated via questionnaires, of which 210 (70%) were returned. Eighty-nine per cent had easy access to a video cassette player. A highly significant number felt that the video would be very helpful or helpful (78%) compared to not helpful, worrying or equivocal 21% (P < 0.0001). This trend was particularly strong in patients < 60 years (83% versus 17%) (P < 0.0001) and those from ethnic groups (95% versus 5%) (P < 0.0001). As a result of this trial, a 20-min film (HEP) has been commissioned. It describes details of the two main treatments for cancer after surgery, namely chemotherapy and radiotherapy, shows patients actually having treatment, and explains the common side-effects and ways to alleviate them. Patients satisfaction with the film and its effect on anxiety and depression are currently being evaluated in an international prospective randomized trial. If it proves advantageous for patients--in view of the ethnic group bias in this study--it will be translated into the ethnic languages of the UK. PMID- 10476111 TI - Two models of care as evaluated by a group of women operated on for breast cancer with regard to their perceived well-being. AB - The main aim of this study was to evaluate two different models of care for breast cancer patients with regard to the patients' perceived well-being. Twenty nine patients were treated in an established care model and 115 patients were treated at a surgical breast clinic with an increased personnel continuity and a short hospital stay. Two questionnaires were used. The Sense of Coherence (SOC) Scale and a study-specific questionnaire concerning perceptions of the hospital stay, information received, body image, social support, pain, health and psycho sociological well-being. Regression analyses were used to study the effects of the care models and the SOC on the patients' perceived well-being. The results showed that the care model with high personnel continuity had a significant positive effect on the patients' emotional state, mental well-being and perception of postoperative pain evaluated 1 year after surgery. The strongest predictors of the patients' well-being postoperatively were their perceived well being before surgery. Furthermore, the stronger the SOC the more positive were the patients' emotional perceptions, perceived general health and mental well being after surgery. The duration of the hospital stay did not show any effect on the patients' well-being. It is concluded that an early preoperative psycho social assessment of the patients is of importance in planning the care, to help patients cope with the disease and its treatment. It is suggested that a care model with high personnel continuity for breast cancer patients could facilitate that assessment. PMID- 10476112 TI - The contribution of molecular genetics to the understanding and management of cancer: potential future applications and implications for nurses. AB - Cancer is an immense medical problem and as a cause of mortality it is second only to cardiovascular disease. Much of the current understanding of cancer is owed to epidemiologists who have discovered a number of causative factors implicated in its development. These causative factors can be divided into genetic, chemical, physical, viral, radiation, immune and hormonal factors. Further advances in understanding have been made over the past decade from contributions made by the field of molecular biology. From investigation and examination of the molecular processes involved in the development of cancer it is becoming increasingly clear that changes in the genetic material of the cell nuclei are the final common pathway to cancer, whatever the initial aetiology. This article will attempt to elucidate the contribution of molecular genetics to the understanding of the likely mechanisms of carcinogenesis, the management of cancer, potential future applications and directions and the implications for nurses arising from this relatively new and evolving field of knowledge. PMID- 10476113 TI - Cerebral metastases in malignant mesothelioma: a case report. AB - A case of a 61-year-old man with metastatic malignant mesothelioma is described. Four months after diagnosis the patient commenced chemotherapy with liposomal doxorubicin as part of an EORTC phase II trial. He developed signs of intracerebral metastases after his fourth cycle of chemotherapy and died shortly after. Malignant mesothelioma is traditionally viewed as a disease that spreads locally but metastasizes rarely. We describe in detail this case and suggest that metastases in this disease are not as uncommon as originally proposed. PMID- 10476114 TI - Public participation in the design of educational programmes for cancer nurses: a case report. AB - Genuine involvement of the public in planning, monitoring and evaluation of health care is a prerequisite to a better quality of service. Similarly users of health services and their carers can contribute much to the planning and delivery of professional education, although this contribution has only recently been acknowledged in any strategic way by nursing educational and professional bodies. This paper provides one example of user and career involvement in the design of continuing education and higher education in cancer care nursing within the University of Leeds School of Healthcare Studies. The author reflects upon the positive aspects and some of the challenges presented in the achievement of public involvement in nurse education. PMID- 10476115 TI - Granulocytic sarcoma of orbit preceding acute myeloid leukaemia: a case report. AB - A rare case of granulocytic sarcoma of orbit preceding the onset of acute meyloblastic leukaemia is presented. A 7-year-old boy presented with proptosis and no other systemic complaints. The appearance of an enlarged muscle in initial tomographic imaging with a normal peripheral blood picture caused a diagnostic problem. Biopsy suggested a diagnosis of granulocytic sarcoma. Subsequent bone marrow study revealed the diagnosis of acute myeloid leukaemia. PMID- 10476116 TI - Caring for people with cancer. PMID- 10476117 TI - Development, implementation, and ongoing monitoring of pathways for the treatment of gastrointestinal cancer at a comprehensive cancer center. AB - The purpose of this article is to explain the process used in the development and implementation of care pathways in a comprehensive cancer center in the Southwest. The pathways are a major component of the disease management process that defines a multidisciplinary standard of practice for a specific episode of care and measure outcomes as a basis for quality and cost improvement. Patients may be on several pathways as they progress through cancer treatment. PMID- 10476118 TI - Photodynamic therapy: a cancer treatment for the 21st century. AB - Esophageal cancer is on the rise and successful treatment continues to be elusive. New treatment modalities are needed to inhibit tumor growth and stop recurrence. This manuscript will highlight photodynamic therapy, a promising new modality for the treatment of esophageal cancer. PMID- 10476119 TI - Education for the gastroenterology cancer patient. AB - Education for the adult gastroenterology cancer patient requires incorporation of standards of care from both an oncology and a gastroenterology focus. In addition, standards of oncology education and role delineations for the gastroenterology/endoscopy nurse provide optimal support for the nurse-teacher working with patients with cancer of the gastrointestinal (GI) tract. The development of a teaching plan and a philosophy for teaching provide the basis for the delivery of care relative to the learning needs of this patient population. PMID- 10476120 TI - Guideline for the use of high-level disinfectants and sterilants for reprocessing of flexible gastrointestinal endoscopes. Society of Gastroenterology Nurses and Associates, Inc. AB - Glutaraldehyde, 0.2% peracetic acid, 7.5% hydrogen peroxide, and 0.08% peracetic acid/1% hydrogen peroxide are cleared by the FDA for reprocessing flexible gastrointestinal endoscopes. Each product has advantages and disadvantages. All require adherence to published reprocessing protocols to maintain the integrity of equipment while providing the public with endoscopic instruments that are safe and effective. All chemicals must be handled with respect. Selection of a product must be weighted against the needs of a particular setting, taking into consideration factors such as compatibility, toxicity, environmental controls, and cost. PMID- 10476121 TI - Kathy's gifts. PMID- 10476122 TI - Making cancer history: a patient's goal. PMID- 10476123 TI - Chemotherapy drugs: an overview for the digestive system. PMID- 10476124 TI - Dynamic aging. PMID- 10476125 TI - The Geriatric Depression Scale (GDS). PMID- 10476126 TI - Elder abuse revisited. AB - 1. There are many types of elder abuse, some of which are never recognized or reported. 2. Medical neglect is a type of elder abuse in health care. 3. Nurses in all health care settings need to be aware of signs and symptoms of elder abuse and take action as indicated. 4. Suspected elder abuse must legally and ethically be reported to the appropriate supervisor and state agencies. PMID- 10476127 TI - Breast cancer in older women: treatment, psychosocial effects, interventions, and outcomes. AB - It is estimated that 48% of women with breast cancer are age 65 and older; yet, scant research focuses on this cohort. Recent advances in breast cancer management have raised many questions regarding appropriate treatment of older patients with cancer. Despite recent emphasis on medical interventions for older women, there is little focus on the unique clinical presentation and psychological sequelae of breast cancer in this population. While elderly women who survive breast cancer seem to cope better than their younger counterparts, their distress must not be overlooked. Interventions such as use of social support, spirituality, and exercise demonstrate therapeutic benefits for older women with cancer and should be explored. Nurses play a key role in educating and supporting older women with breast cancer and can help dispel many age-related myths and misconceptions. PMID- 10476128 TI - Physical and pharmacologic restraints in long-term care facilities. AB - This study examined the effects of education on the attitudes and practices of long-term care staff toward use of restraints. The intervention, a 1-day educational seminar, used a collaborative team of speakers from the Utah Survey Agency and medical professions. Seminar goals were threefold: first, to provide information about best practices for managing behaviors of individuals with dementia in long-term care settings; second, to provide an explanation of the Omnibus Budget Reconciliation Act regulations pertaining to restraint use; and third, to present alternative strategies to link best practice guidelines to the provision of care. Results showed significant changes in participants' attitudes toward use of restraints. Participants reported replicating the seminar for nursing home staff, revisiting facility policies on restraints, and modifying resident care plans. PMID- 10476129 TI - Pain assessment in older adults who are cognitively impaired. PMID- 10476130 TI - Comparison of three instruments in predicting accidental falls in selected inpatients in a general teaching hospital. AB - Accidental patient falls are becoming a major cause of concern for hospitalized inpatients. It is well known that patients who fall once during their hospital stay are more likely to fall again and that fall rates tend to be higher in hospitalized elderly individuals. Concerned health care team personnel recognize that many accidental patient falls may be predicted and, thus, prevented. The best tool to predict falls has not been determined yet. The purpose of this study was to compare the abilities of the Morse Fall Scale (MFS), the Functional Reach (FR) test, and the nurses' clinical judgment in predicting those inpatients on a rehabilitation unit and a geriatric medical ward who were most likely to fall. A total of 98 patients were screened in a 3-month period, with each patient undergoing all three instruments the same day. The results showed that the two objective standardized tests (i.e., MFS, FR) were time consuming and often inconvenient and were no better at prediction than the clinical judgments made by the primary nurses. PMID- 10476131 TI - What types of clinical experiences do you think should be included in geriatric nursing courses at the undergraduate level? PMID- 10476132 TI - State of the society. PMID- 10476133 TI - Presidential address. Welcome to the future! PMID- 10476134 TI - Home intravenous therapy down under. AB - Health service research traditionally has relied on quantitative methodologies that view the question from the researcher's perspective. Exploring the question of what it is like to receive intravenous therapy at home has required investigation into what is known about home i.v. therapy and the impact on the patient in particular. The purpose of this study was to describe and understand the recipient's lived experience of home i.v. therapy using a research approach that preserves the uniqueness of the experience from the recipient's perspective. The method involved conducting in-depth interviews with a sample of 26 recipients of home i.v. therapy throughout Tasmania, Australia. Interviews were analyzed using an interpretative process, with the participants describing a process that included the main themes of "having a life" and "the hazards of hospitalization." The main goals of the work are to heighten nurses' and other healthcare professionals' understanding and awareness of home i.v. therapy from the consumer perspective and to address the lack of patient/consumer advocacy and exploration within the evaluative literature available on this subject. PMID- 10476135 TI - Developing a protocol to prevent and treat pediatric central venous catheter occlusions. AB - Catheter occlusions are a common occurrence in pediatric patients with central venous catheters. These occlusions are attributable to many factors, such as mechanical problems caused by catheter and patient size, clot formation caused by blood product administration and laboratory sampling, drug precipitation, and lipid residues. Because of the significant patient risk and cost involved in replacing occluded central venous catheters, a multidisciplinary team used a quality improvement approach to determine the effectiveness of urokinase in pediatric patients. Data from the project enabled development of a decision tree for urokinase use. Statistics from the pilot test revealed that 85% of the catheter occlusions were related to thrombotic events, and urokinase was effective in all cases. However, 15% of central line occlusions were related to medication precipitates and were not effectively cleared with urokinase. Given the evidence that urokinase is effective only on fibrin-related occlusions, the decision tree was modified to improve success with occlusions caused by medication precipitates. PMID- 10476136 TI - Patient confidentiality in the electronic age. AB - This article presents an overview of confidentiality issues as they relate to patient information stored or disseminated electronically. The three related issues of confidentiality, privacy, and security are defined, and how those issues have changed with the advent of computerization is discussed. Topic discussed are the risks versus the benefits of computerizing; threats to the organization and the patient; and the legal, organizational, and technological solutions that can and should be implemented to ensure all patient-related information is kept confidential and secure. PMID- 10476137 TI - Intravenous immunoglobulin use in pediatric patients. AB - New trends in home intravenous therapy for the pediatric population are being developed. One trend in particular is proving to be both exciting and promising. This trend is the use of intravenous immunoglobulin in the homecare setting. This article explores the most commonly seen disorders and treatments using this specific intravenous prescription. Safe nursing practice and future trends also are discussed. PMID- 10476138 TI - Intravenous push medications in the home. AB - The intravenous push method for drug administration is not a new idea. In an era of quality, good patient outcomes, and cost efficiency it is important to look at economical and safe methods of drug delivery. This article reviews different methods of drug delivery and compares outcomes. The focus of the study was a comparison of minibag versus i.v. push drug delivery for anti-infective self administration in the home setting. Case reports totalling 1116 from 50 sites were analyzed. The most frequently used drugs were ceftriaxone, cefazolin, and ceftazidime. Rates for all complications were 1.89/1000 catheter days for i.v. push and 1.69/1000 days for minibag delivery method. The phlebitis rate was 0.6/1000 catheter days for i.v. push and 0.79/1000 catheter days for minibag delivery method. Client satisfaction rates were comparable for all types of drug delivery methods. PMID- 10476139 TI - Tuberculosis. An overview. AB - This article provides an explanation of the emergence of multidrug resistant tuberculosis. Combination drug therapy, early identification, isolation, treatment, complication issues, and direct observation therapy are discussed. Infection control measures, including the importance of properly fitting personal protective equipment and routine testing of healthcare workers, are reviewed. PMID- 10476140 TI - Cross-cultural nursing: the cultural perspective. PMID- 10476141 TI - Transcultural versus cross-cultural. PMID- 10476142 TI - Transcultural nursing at Y2K: some thoughts and observations. PMID- 10476143 TI - What is transcultural nursing and culturally competent care? PMID- 10476144 TI - Cultural openness: intrinsic to human care. PMID- 10476145 TI - Cultural competence and psychiatric-mental health nursing. PMID- 10476146 TI - Culturally competent care. PMID- 10476147 TI - Global health awareness and transcultural relationships. PMID- 10476149 TI - Infant feeding styles of West Indian women. AB - An ethnographic field study design was used to explore infant feeding among 20 West Indian women on the island of St. Croix, United States Virgin Islands. Two styles of infant feeding emerged from the data, "older style" and "contemporary style." Three patterns of combined breast and bottle feeding were identified: day/night, supplemental, and random. Older style mothers followed a day/night pattern, whereas contemporary style mothers used a supplemental pattern. A folk explanatory model of infant feeding was constructed and included why infants were fed in certain ways, as well as the meaning of feeding practices. The health care system had an impact on both styles of infant feeding through hospital practices and provision of formula through the Women, Infants, and Children (WIC) program. PMID- 10476148 TI - Engaged mothering: the transition to motherhood for a group of African American women. AB - Using grounded theory methodology, 17 first-time African American mothers were interviewed to elicit their experiences of pregnancy and motherhood. Participants had a mean age of 30 years, were mostly married, employed, middle income, college educated, and all received adequate prenatal care. Engaged Mothering was identified as the core category, denoting the active, involved, and mutual process in which a woman prepares to be a mother, cares for herself and her infant, and dreams about and plans for the future to have a good life for her child. Strategies women used in this process included getting ready, dealing with the reality, settling in, and dreaming. Conditions of intentionality of the pregnancy and prior history of miscarriage or health problems of the mother affected the process. Women described the effects of racism on their daily lives and on the criteria they used to choose providers. Nursing interventions are proposed based on these results. PMID- 10476150 TI - Meanings and practices of health among married Thai immigrant women in Sweden. AB - The aim of this qualitative study was to explore and explain the meanings and practices of health among Thai immigrant women in Sweden with Swedish husbands. In agreement with Leininger's theory of cultural care, data were collected through observation-participation and ethnographic interviews with 10 key informants and 16 general informants living in Uppsala. The principal meanings of health of the Thai immigrant women were abstracted from the raw data using structural analysis. They were found to be (a) state of well-being, (b) absence of illness, (c) ability to perform daily role activities, and (d) adaptation to their new life situation. The principal factors for the promotion of health among these women were (a) mental support from family, (b) nutrition, (c) physical exercise, (d) no smoking, and (e) modest drinking. Knowledge among health care professionals of meanings and practices of health, such as those found here, is vital for the provision of culturally congruent care in a multicultural society such as Sweden. PMID- 10476152 TI - The meaning and experience of health ministry within the culture of a congregation with a parish nurse. AB - A growing number of nurses are working with congregations in churches, synagogues, and mosques, trying to promote health through integrating religious beliefs and health knowledge. The concept of health ministry guides nursing practice when working with congregations. This ethnographic study aims to understand the meaning and experience of health ministry within the culture of a congregation with a parish nurse examining both emic and etic perspectives. Utilizing participant observation, interviews, and reviews of written documents, an ethnography was created of an ethnically diverse, urban United Methodist congregation with a volunteer parish nurse. Two forms of health ministry were found within the congregation, which the investigator named extrinsic and intrinsic health ministry. Extrinsic health ministry included activities whose explicit purpose was to promote health. Intrinsic health ministry included activities and experiences within congregational life whose express purpose was something other than health promotion, yet participants identified it as promoting their health. Implications for transcultural nurses include using knowledge of congregational cultures to facilitate culturally congruent health ministry. PMID- 10476151 TI - Culture care meanings and experiences of postpartum depression among Jordanian Australian women: a transcultural study. AB - This study discovers, describes, and explains the personal experiences, perceptions, and care meanings of Jordanian women who have suffered postpartum depression. Most postpartum cases often are misdiagnosed as exclusively psychological and untreated by health care professionals without consideration to the cultural meanings of this problem. Understanding the experiences of these women is important, as their expressions often are contextually and culturally influenced. Using Leininger's Theory of Culture Care Diversity and Universality, a purposive sample of 22 Jordanian women diagnosed with postpartum depression, living in Sydney, were interviewed. The ethnonursing research method and data analysis procedures were used. Results revealed that Jordanian mothers experienced severe loss of control over emotions of loneliness, hopelessness, and feelings of being a bad mother. Three major themes focusing on the care meanings and experiences of Jordanian women are discussed: (a) Care means strong family support and kinship during the postpartum period, (b) care is carrying out and fulfilling traditional gender roles as mother and wife, and (c) care is preservation of Jordanian childbearing customs as expressed in the celebration of the birth of the baby. PMID- 10476153 TI - Faculty experiences teaching Native Americans in a university setting. AB - Nursing faculty told their stories about the beginnings of a Native American Family Nurse Practitioner Recruitment Program. Through hermeneutical analysis, the authors' findings reveal a strong academic worldview, active in maintaining professional standards. This traditional view tends to override efforts to provide individualized programs that are culturally relevant to the Native American worldviews. By understanding the value conflicts active in this study, nursing faculty can begin dialogue to create new learning experiences that are more culturally relevant. PMID- 10476154 TI - Pica: sorting it out. AB - Pica, a culture-bound illness, has occurred for centuries. The ingestion of nonfood substances such as starch, cornstarch, clay, dirt, and other material is fairly common, although the distribution of the condition varies by cultural and socioeconomic factors. The underlying cause of pica is not known, although the condition often is associated with pregnancy. There is conflicting evidence about the association of nutrient deficiencies and pica. This article presents a clinical example of pica in a pregnant 33-year-old African American woman. Implications for culturally appropriate care are discussed. PMID- 10476158 TI - Founder's focus--transcultural nursing leadership: vital for the third millennium. PMID- 10476155 TI - How to search for information on transcultural nursing and health subjects: Internet and CD-ROM resources. AB - With the proliferation of electronic resources available to search for subjects related to transcultural nursing and health, nurses must keep abreast of computer based tools that enable them to quickly and efficiently obtain information on a variety of topics. This article provides suggestions for narrowing and focusing a search on transcultural nursing and related subjects using key terms indexed in the Cumulative Index to Nursing and Allied Health Literature, International Nursing Index, Medline, Index Medicus, and Psychological Abstracts/PsychLit. Information about accessing the Native American Resource Information Service, Education Resources Information Center, Infotrac/Expanded Academic Index, and Sociological Abstracts also is provided. In the article, selected examples of Internet sites of interest in transcultural nursing and health are identified with their universal resource locator, and they are annotated. Web sites for U.S. government agencies, organizations, and commercial groups that concern transcultural nursing and health are cited. Global transcultural health and nursing Internet resources also are included. PMID- 10476159 TI - "Ethnic cleansing": the ultimate tragedy of intolerance. PMID- 10476160 TI - Multimeasure pain assessment in an ethnically diverse group of patients with cancer. AB - The purposes of this study were (a) to describe the relationship between pain perception and ethnic identity and socioeconomic status, (b) to evaluate the intercorrelations between pain measures in different ethnic groups, and (c) to determine whether ethnicity or socioeconomic status influences patient's pain control beliefs and satisfaction with the pain management provided. The sample consisted of 51 English-, Korean-, or Spanish-speaking participants experiencing cancer pain who were 18 years and older and were having a Karnofsky score of no less than 30. The Visual Analogue Scale (VAS), Memorial Pain Scale (MPS), and Face Scale (FS) were used to measure pain perception. In all pain analyses, Hispanics, African Americans, and Anglos did not differ significantly. The data suggest that the pain scales used in this study are appropriate for use in a multicultural population. PMID- 10476161 TI - Hearing our voices: assessing HIV prevention needs among Asian and Pacific Islander women. AB - The purpose of this study was to (a) assess the impact of HIV/AIDS on the Asian Pacific Islander (API) community and changes in their behavior due to AIDS, (b) identify perception of risk, HIV risk behaviors, factors contributing to those behaviors, barriers to HIV prevention, and the types of prevention programs that would benefit their community, and (c) describe culturally appropriate considerations when designing HIV prevention strategies for API women. Thirty API adults participated in three different groups. Focus group interviewing methods were used, guided by the Health Belief Model. The women had numerous concerns about HIV that placed them at risk for infection, such as their inability to talk with their sexual partners about condom use due to the cultural and taboo nature of sexual topics. All groups concluded that for HIV prevention interventions to be successful, they must be tailored to the cultural and specific needs of API women. PMID- 10476162 TI - Transcultural nursing study of emic and etic care in the home. AB - This study describes 18 informants' experiences with home care by multidisciplinary health care providers using participant observation as research methodology. Study findings support the salience of culture as the context for interpreting, valuing, and defining satisfaction with care. In contrast to hospital care, home care was perceived by informants as enhancing their ability to remain in their valued setting, supportive of their restorative and health maintenance needs. In particular, all informants viewed their home as the milieu that allows for continuity of their lifestyles, family relationships, and cultural values. PMID- 10476163 TI - Perinatal death: uncovering the needs of midwives and nurses and exploring helpful interventions in the United States, England, and Japan. AB - Perinatal death is a crisis for midwives and nurses as well as for bereaved parents and extended families. Surveys and interviews conducted in the United States, England, and Japan described the needs and responses of nurses and midwives as they coped with their own feelings while caring for bereaved parents. Results emphasized common needs of caregivers for increased knowledge, mentored experience, communication skills, and personal support to confidently provide sensitive care to families. Although need for education regarding cultural specific care was revealed, participants identified helpful strategies of care for bereaved parents that could extend and improve care universally. PMID- 10476164 TI - The essence of Native American nursing. AB - This article emerged at the "Gathering of Wisdom: American Indian Nursing Summit III," which took place in October 1997 on the Flathead Indian Reservations in Polson, Montana. This gathering described and clarified the distinctiveness of Native American nursing. Nursing literature has explored the essence of nursing from a variety of perspectives. This article describes a process of discovery in which seven themes emerged on the essence of Native American nursing. These themes, in ascending order of significance, are caring, traditions, respect, connection, holism, trust, and spirituality. These themes are connected and interwoven throughout the center of Native American culture and can provide a creative outlook for transcultural nursing and nursing in general. Furthermore, these themes provide a vehicle for care delivery innovation. PMID- 10476165 TI - The use of folk healing and healers by six Latinos living in New England: a preliminary study. AB - The purpose of this qualitative study is to describe Latinos' use of folk medicine and the values associated with it in the context of mainstream health care. The research questions focused on the meaning of folk medicine in the Latino culture and how participants made decisions regarding the use of folk medicine or modern biomedicine. Interviews were conducted with six Latinos living in a New England city who had used folk medicine. Three themes were identified: being sick, healers and healing, and faith and believing. The participants in this study valued their cultural folk medicine and folk healers. However, none relied exclusively on folk medicine. All used mainstream health care providers for specific purposes. A major reason for not using mainstream health care providers more frequently was the perceived lack of holistic care and the use of medicines that are not natural. PMID- 10476166 TI - The Cummins model: a framework for teaching nursing students for whom English is a second language. AB - The health care system requires nurses with the language ability and the cultural knowledge to meet the health care needs of ethnic minority immigrants. The recruitment, admission, retention, and graduation of English as a Second Language (ESL) students are essential to provide the workforce to meet the demands of the multicultural community. Yet, ESL students possess language difficulties that affect their academic achievement in nursing programs. The application of the Cummins Model of language proficiency is discussed. The Cummins Model provides a framework for nursing faculty to develop educational support that meets the learning needs of ESL students. PMID- 10476168 TI - President's message. PMID- 10476167 TI - Utilising culturally congruent strategies to enhance recruitment and retention of Australian indigenous nursing students. AB - Recruitment and retention of Aboriginal and Torres Strait Islander (ATSI) nursing students has been the concern of many faculties of nursing across Australia for some time. Multiple factors and issues have been raised to address recruitment, and most important retention, of ATSI students in undergraduate nursing programs. This article, through a review of the literature, explores and describes discoveries and discusses the importance of culturally meaningful strategies and knowledge as significant in addressing this core issue. Strategies for change in relation to curriculum design and faculty education in transcultural nursing are described. PMID- 10476169 TI - AT&T language line: educating transcultural nurses on federal mandates and interpreter use. PMID- 10476170 TI - Y2K: are you ready? PMID- 10476171 TI - Are we ignoring the lessons of history? PMID- 10476172 TI - Navigating the literature. PMID- 10476173 TI - Clinical practice guidelines for pressure ulcer prevention can prevent malpractice lawsuits in older patients. AB - OBJECTIVE: To evaluate the impact of implementation of and compliance with practice guidelines for pressure ulcer (PU) prevention using medical malpractice litigation data. SETTING AND SUBJECTS: Forty-nine plaintiffs whose respective compensations ($14,418,770 in 35 plaintiffs) or dismissals had been reported in 2 legal databases. PU verdicts and settlements for plaintiffs 60 years of age and older were evaluated using the American Geriatric Society's Clinical Practice Guidelines, "Pressure Ulcers in Adults: Prediction and Prevention." METHODS: Litigation analysis was used to identify the effect, implementation of, and compliance with PU prevention practice guidelines on malpractice awards in PU lawsuits. Data were obtained using fact patterns from 2 legal databases, LEXIS and WESTLAW. Potential decreases in plaintiff awards and prevention of disability were calculated assuming that health care defendants had modified their behavior to conform to the practice guidelines. Possible increases in defendant awards were used to estimate the added risks to health care professionals of adopting these guidelines as the standard of care. MAIN OUTCOME MEASURES: Projected changes in verdicts, monetary awards expressed in dollars, and disability score. RESULTS: Had health care defendants followed these guidelines, $11,389,989 might have been saved in 20 lawsuits. Violations of guidelines appeared to "cluster" together, with many plaintiffs alleging breeches of several interrelated guidelines. It appears that improving the level of care required to remedy 1 guideline variation could improve the outcomes for the entire cluster. In contrast, the use of the guidelines in court as the standard of care against defendant health care professionals might have contributed to changing only 4 of 14 defense verdicts. CONCLUSIONS: Use of clinical pathways in these settings can benefit both caregivers and patients by favorably modifying preventive practice patterns while decreasing vulnerability to litigation. Conversely, the continuing threat of fault-based litigation against substandard practitioners and facilities provides an ongoing safeguard of patient rights and reduces the risk of subsequent disability. PMID- 10476174 TI - The lived experience of having an ileoanal reservoir: a phenomenologic study. AB - OBJECTIVE: Ileoanal reservoir (IAR) surgery or ileal pouch anal anastomosis is a relatively new surgical technique for people with ulcerative colitis or familial adenomatous polyposis. Little attention has been given to the experiences of people undergoing the procedure. The purpose of this qualitative study was to understand the lived experience of persons who have had construction of an IAR. DESIGN: The study was grounded in the phenomenologic approach of Van Manen to capture the lived experience of having an IAR as perceived by those persons in their everyday world. SETTING AND SUBJECTS: A purposive sample of 10 individuals with direct and personal knowledge of the experience were interviewed in detail. Subjects were recruited by contact with an IAR support group and a local WOCN group. All interviews were conducted in a private setting in a health care facility, school, or the participant's home. METHODS: Data collection involved face-to-face interviews lasting from 1 to 2 hours. Interviews were audiotaped and transcribed. MAIN OUTCOME MEASURES: Demographic data from the interviews were tabulated. Analyses of transcripts revealed 10 essential thematic categories with multiple theme clusters of the IAR experience related to lived body, time, space, and relationships. Analyses were completed by the researcher and a faculty mentor. RESULTS: The particular thematic categories that evolved from the data were (1) restricted life world, (2) living with uncertainty and fear, (3) seeking control, (4) vicious cycles: crisis and normalcy, (5) seeking and giving support, (6) alienation from the body, (7) living with bodily alterations, (8) the gift of time, (9) role and relationship changes, and (10) the end of the tunnel but relative results. All 10 theme categories were correlated with a literature review and other sources. CONCLUSION: The study provides a portrait of courage and survival for individuals experiencing major surgical interventions and bodily invasion associated with IAR after years of living with UC. Implications for nurses and health care personnel and questions for future research are presented. PMID- 10476175 TI - Ileoanal procedures: acute and long-term management issues. AB - The pelvic pouch procedure has become the preferred surgical treatment for patients with ulcerative colitis and familial adenomatous polyposis. Even though this operation removes the diseased colon and allows defecation through the anus, the outcome is not "perfect." Rather, ongoing management issues occur that require attention. This article reviews acute and chronic management issues related to the creation and management of the pelvic pouch. PMID- 10476177 TI - Descriptive basics: measures of central tendency. PMID- 10476176 TI - Use and costs of incontinence pads in female study volunteers. Continence Program for Women Research Group. AB - OBJECTIVES: We examined the use and cost of incontinence pads and the relationship to factors such as age, duration of incontinence, diurnal frequency, incontinence severity indices, urodynamic diagnosis, and quality of life. SUBJECTS AND SETTING: Three hundred fifteen women with urinary incontinence who volunteered to participate in 1 of 3 incontinence studies (behavioral intervention, estrogen supplementation, or surgery) were analyzed. Subjects were community-dwelling women aged 45 years and older living in 3 cities in the southeastern United States. METHODS: Pad use was recorded on a daily diary. The type of pads used was reported on the history. Average price of pad types was assessed at local stores and reported in 1995 dollars. Statistical comparisons used nonparametric methods. MAIN OUTCOME MEASURES: The number of pads used per week and annual cost of pads in 1995 dollars. RESULTS: Seventy-seven percent of subjects used pads at baseline. Median cost per year for the entire cohort was $46 (interquartile range $3-$138). For pad users, median annual cost was $76 (interquartile range $36-$177), with costs being greater for women with detrusor instability than those with pure genuine stress incontinence (median $135-$138 versus $63). This increased cost was likely associated with the greater use of special incontinence products among women with detrusor instability. For the entire cohort, cost and usage did not differ by urodynamic diagnosis. Cost and pad usage were significantly associated with number of incontinent episodes and quality of life, but not with age, pad weight, or duration of incontinence. CONCLUSIONS: The majority of incontinent women who sought treatment used absorbent pads at least once per week, with menstrual pads being the most common type of pad. The annual cost of pad usage was not as high as in previous estimates. PMID- 10476178 TI - Management of a neonate with necrotizing entero-colitis and eight prolapsed stomas in a dehisced wound. PMID- 10476179 TI - What have you done for your customers lately? PMID- 10476180 TI - Communication: an outcome of case management. AB - Case management is a healthcare delivery model emphasizing coordination and networking of patient care services across multidisciplinary providers and various care settings. This requires effective communication to overcome traditional barriers to information exchange. This study investigates communication as an outcome of case management. The classic communication model of sender-message-channel-receiver is used as a conceptual framework. Using a closed medical record review, the amount and type of referral information exchanged between a hospital using a case management model to coordinate care and a home health agency accepting the case-managed patient is described. In addition, the influence of selected organizational and medical condition factors on communication is explored. PMID- 10476181 TI - Developing case management in a nursing home environment. PMID- 10476182 TI - Integration of nutrition screening into case management practice. AB - Nurse case managers who are responsible for identifying the needs of those requiring case management intervention are ideal professionals to recognize patients who are at risk of developing malnutrition or who are already malnourished. The primary objective of case management is to provide proactive, comprehensive assessment of individuals before a preventable health crisis occurs. The objectives of nutrition screening complement those of case management because the underlying benefit to the organization and the patient is the decrease in morbidity and mortality rates, length of stay, and cost. This article provides information on nutrition screening and its impact on patient outcomes, techniques for integrating nutrition screening into case management practice, tools for conducting nutrition screening, and information that may be helpful in communicating nutrition-related needs to providers and other healthcare professionals. PMID- 10476183 TI - Case management program for breast cancer education. PMID- 10476184 TI - Pain among the elderly: an action plan. AB - Pain is the most frequently communicated nursing-sensitive patient outcome among the elderly. An effective action plan to decrease pain among the elderly should encompass both medically based interventions and psychosocial techniques. Nurses who are familiar with the latest information about pain and who are cognizant of and comfortable with their own views about pain are likely to bring about a positive patient outcome. Further research will add to the current knowledge base while laying the groundwork for new interventions. PMID- 10476185 TI - Ethical dilemmas in artificial nutrition and hydration: decision-making. PMID- 10476187 TI - Caring sciences. PMID- 10476186 TI - Team case management of chronically mentally ill veterans: a group therapy approach. AB - This article describes a team case management model using a group therapy approach developed and implemented for chronically mentally ill veterans. The model was developed to address inadequacies in the system and to provide mentally ill veterans with more frequent interventions and an organized support system to maintain them in the community. Two hundred seventy-five patients enrolled in the case management system attend weekly groups led by two or three interdisciplinary staff. In addition to serving as primary therapists and case managers, the interdisciplinary staff broker services, linking patients with needed services within the Veterans Affairs system and the community. The results indicate that the case management model has improved the lives of those enrolled by improving quality of life, medication and treatment compliance, social relationships, and general well-being. Use of emergency room services has decreased, as well as psychiatric symptomatology. For some diagnostic categories, the number of hospital readmissions and days hospitalized also decreased. PMID- 10476188 TI - Patients' fears in coronary arteriography. AB - This study was concerned with the objects of fear and the intensity of fears related to coronary arteriography (CA) in 378 patients scheduled for coronary arteriography at a university hospital. Intensity of fear was measured on a 10 point Likert-type scale which listed 26 objects of fear. The patients were asked to assess the intensity of their fears both before and after CA. The results showed that the intensity of fear varied depending upon the object of fear. The highest intensity was recorded for fear of coronary artery bypass surgery. A significant increase was found in fear of lying flat in bed after CA and fear of not receiving social support. Fears of uncertainty about the illness, the CA procedure, results of CA, pain, coronary artery bypass surgery and coronary angioplasty decreased significantly after CA. Significant associations were found between objects of fear and patient demographics. It is concluded that patients' fears should be assessed individually both before and after CA. Special attention should be paid to the fears of women, patients under 45 years, upper and lower level employees and the unemployed. PMID- 10476189 TI - Patient-perceived quality of life after coronary bypass surgery. Experienced problems and reactions to supportive care one year after the operation. AB - In an interview conducted one year after coronary bypass surgery (CABG), 115 patients (92 males, 23 females) were asked to discuss their present quality of life, problems related to the operation and their reactions to the supportive care following the operation. The main purpose of this study was to assess whether the patients' own presentation of these questions revealed information not elucidated through a standard questionnaire which included the Nottingham Health Profile and other questions on physical, emotional and social well-being. About one quarter of the interviewed patients expressed dissatisfaction concerning their present quality of life, information which corresponded well with the findings in the questionnaires. Reported limitation in physical activity did not, however, match exercise test findings. The interviews alone revealed additional information concerning well-being. Most important were problems with wound healing and difficulties describing and interpreting pain and other discomfort in the thoracic area. Several patients also expressed dissatisfaction with the postoperative supportive care. We conclude that there is a need for a complementary approach to estimate quality of life, and that this could be included in an extended nursing care program designed to meet the varying needs of patients after CABG. PMID- 10476190 TI - Structure and content in Norwegian nursing care documentation. AB - In 1994, the Norwegian Board of Health (NBH) published recommendations for nursing care documentation. The two-fold purpose of the present study was to see if 5 wards in 2 Norwegian hospitals fulfilled the proposed NBH recommendations and guidelines regarding documentation, and to evaluate them in terms of the proposed structure and key words of the VIPS model. Results showed that all nursing records (n = 55) had an admission assessment. A nursing care plan was present in 62% of the records. Nursing goals were lacking in the remaining 38%, diagnosis and planned interventions were absent in 18%, and 45% of the diagnoses lacked information concerning patient progress or outcome. The nursing care plans were updated in only 40% of the records and discharge notes were present in 35%, confirming that NBH recommendations were not met in this sample. The key words of the VIPS model covered all information present in the records, and high interrater reliability was obtained for the majority of key words categorized by two independent researchers. It is suggested that the VIPS model components and key words can contribute to a reliable and uniform model for nursing care documentation and enhance comprehensive and systematic documentation, which is presently lacking in Norwegian records. PMID- 10476191 TI - Problem-focused coping and satisfaction with activities of daily living in individuals with muscular dystrophy and postpolio syndrome. AB - The problems, problem-focused coping and satisfaction with activities of daily living in a total of 119 persons with muscular weakness are described. The study encompasses three groups: 33 persons with various types of muscular dystrophy, 46 with myotonic dystrophy and 40 with symptoms relating to the post-polio syndrome. A self-report instrument for Assessment of Problem-focused Coping (APC) was used. Most of the problems reported were connected with mobility and transportation and work, but the subjects used problem-focused coping relatively infrequently and few differences were found between the different groups or between the sexes. The most widely used problem-focused coping strategy was 'Devices and tricks'. The highest degree of satisfaction was noted for personal care and the lowest for mobility and transportation. The APC provides information about patients' evaluation of everyday situations, forming a complement to functional measurements in the field of rehabilitation. PMID- 10476192 TI - Pain in persons with post-polio. The Swedish version of the Multidimensional Pain Inventory (MPI). AB - Increasing muscular atrophy and joint instability in the post-polio syndrome (PPS) leads to muscle and joint pain. The aim of this study was to describe how persons with post-polio syndrome (PPS) perceive their pain and how the pain affects their everyday lives. The Multidimensional Pain Inventory (MPI-S) was used together with supplementary questions concerning location and verbal description of the pain. The study group comprised 37 persons with PPS who had had pain for an average of 19 years. Most commonly reported was pain from the joints of the extremities, followed by pain from the lower back. The verbal description suggests that pain is a palpable health problem for this study group. The results of the MPI-S show that women had the most pain and that the younger the persons are, the more the pain involves negative stress experiences. Activities such as outdoor work, social and other activities away from home involve difficulties, according to the results. The study group could manage their pain, experienced social support, and especially women managed to carry out household chores despite their pain. The MPI-S demonstrates acceptable reliability for this study group. PMID- 10476193 TI - Nursing interventions in crisis-oriented and long-term psychiatric home care. AB - Psychiatric nurses in The Netherlands are moving out of residential mental health institutions and are pioneering home care for the acutely and chronically mentally ill. The purpose of this study was to identify the interventions nurses currently use and to describe the differences between crisis-oriented and long term psychiatric home care. Data was collected of 159 nursing care plans from four participating crisis-oriented and two long-term psychiatric home care teams. All stated nursing activities were identified and subsequently labelled and classified using the Nursing Intervention Classification (NIC). Results revealed that in both crisis-oriented and long-term psychiatric home care, nurses used a wide range of nursing interventions. Medication Management, Coping Assistance and Activity Therapy were the most frequently undertaken nursing interventions in both types of care. Within crisis-oriented care, Emotional Support and Self esteem Enhancement dominated, whereas long-term care focused on Socialization Enhancement and Home Maintenance Assistance. The results will be used for further research and for standardization of nursing care plans within these categories of nursing practice. PMID- 10476194 TI - Recurrent infections and musculoskeletal disorders--signs of generally increased care demands? A descriptive study of medical records. AB - In this study we describe and analyse the health status, health-care consumption and the characteristics of persons visiting the primary care centre because of frequent symptoms of respiratory tract infections (RTI) (n = 83) and disorders related to the musculoskeletal system (DMS) (n = 166) as described in medical records for a three-year period. When all the medical diagnoses, health-care consumption, sick leave and other information had been gathered, the findings were that more women (73%) than men fulfilled the inclusion criteria. On average, the group with the highest frequency of symptoms paid 8.3 visits/three years to the physician compared with 1.0 for the controls (p < 0.05). The average total sick-leave attributable to RTI was 29.4 days/3 years and 8.1 days/3 years for the controls. Forty percent of patients in the frequent symptoms group had regular contact with children at work compared with only 18% of the controls. Subjects in the former group were usually manual workers with an increased physical and psychological workload compared with the controls. This group of subjects probably represents a category of vulnerable patients and their total situation should be further investigated. PMID- 10476195 TI - Senses of responsibility. A challenge for occupational and physical therapists in the context of ongoing professionalization. AB - The aim of this paper was to delineate different senses of responsibility in the therapeutic relationship between occupational and physical therapists and their patients, in the context of ongoing professionalization. To assist the analysis of the therapists' responsibilities, Ozar's guild and interactive relationship models were employed. The questions with regard to each model asked how the model might be used in order to describe the therapists' professional development in general, and specifically their professional responsibilities. Based on earlier findings, the professional development of the therapy groups has developed in line with both the guild and interactive models, i.e. some therapists have adopted the former and others the latter. Whether therapists in general, in recent years, have shifted their paradigm, and consequently also their choice of relationship model, is a question that necessitates further research. However, based on the results from the analysis here, it seems relevant to propose that therapists, with a renewed sense of urgency, should use Ozar's models as tools for reflection on the development of their professional responsibilities in the therapist-patient relationship, in the frame of a rehabilitation paradigm. PMID- 10476196 TI - Assessing the core of nursing. PMID- 10476197 TI - Assessing your publication potential. AB - To be published in a professional nursing journal is an admirable goal and one that is obtainable if you have skills, knowledge, and tenacity and if you want to share information with others. Everything we do or accomplish in life is built on steps, and that translates into taking one step after another. Writing for publication is no different; this article enumerates and describes the 11 steps to becoming published. Self-answer questions direct the reader to consider whether to compose solo or with a co-author(s). The nursing journal review processes and options of where to submit queries or completed articles are covered. Books to help writers begin writing or refining their work are included. This article states that the first rule of becoming published is "where there's a will, there's a way." It concludes with the dictate, start now. Procrastination has stifled more projects than any other obstacle. Pick up your pen or turn on your computer and go to work. PMID- 10476198 TI - Postoperative pain management in the non-Hispanic white and Mexican American older adult. AB - The purpose of this study was to assess use of pain relief measures and satisfaction with postoperative pain management among non-Hispanic white and Mexican American older adults after abdominal surgery. Findings revealed interpatient and intraethnic diversity, endorsing the importance of careful patient assessment and examining patient satisfaction for quality assurance/improvement. PMID- 10476199 TI - Assessing graduate perioperative education. AB - You may be one of many perioperative nurses who are considering graduate education. Although there are some obstacles to this goal, they are not insurmountable. This article examines the motivators and obstacles to master's level nursing education. The educational options, assessment criteria, and admission process are described. PMID- 10476200 TI - Assessing the practicality of research findings for clinical practice. AB - Reading research articles can be challenging, but it also can be stimulating. Once you get a system for reviewing articles and interpreting their worth and value to your practice, you will find much usable information. Reviewing articles with the focus of assessing their importance to clinical practice could be done with a research committee of your local AORN chapter. It also would be an interesting and informational chapter program to have a panel discuss the relevance of two or three studies to perioperative practice. This process and type of information also can be used if your institution is developing research teams. Assessing the practicality of research to your practice is essential. Research is not a mystery; it is the answer to a mystery. It is necessary to the future of perioperative nursing. Change practice with empirical evidence! PMID- 10476201 TI - Infection control assessment in the perioperative setting. AB - Perioperative nursing staff are continually involved in various forms of assessment related to patient care. Inherent in the surgical setting is the effort to minimize the risk of nosocomial infection to the patient and, at the same time, provide a safe environment for the health care professional. Perioperative staff must consider the multiple factors that influence infection outcomes and risks in the surgical setting and be familiar with strategies for personal protection against communicable disease. PMID- 10476202 TI - Assessment: use it to establish an outpatient clinic. AB - If faced with the task of establishing an adult genitourinary outpatient clinic in an already existing ambulatory surgery setting, would you know where to begin? Careful, considered assessment is crucial to a successful outcome. PMID- 10476203 TI - Preoperative nursing assessment of the adult patient. AB - The preoperative patient assessment must be very focused, providing detail and specificity in several essential assessment areas. The preoperative patient assessment helps to determine whether the patient is a high surgical risk. Patients who are determined to be high surgical risks are at a higher risk for perioperative and postoperative mortality and morbidity. PMID- 10476204 TI - RNA repair as a novel approach to genetic therapy. PMID- 10476205 TI - Politically correct gene therapy? A "clean environment" improves gene delivery to the brain! PMID- 10476206 TI - Utilization of variant-type of human alpha-fetoprotein promoter in gene therapy targeting for hepatocellular carcinoma. AB - We previously reported that the retroviral vector (LNAFW0.3TK) expressing the herpes simplex thymidine kinase (HSVtk) gene under the control of the 0.3 kb human alpha-fetoprotein (AFP) promoter provided the ganciclovir (GCV)-mediated cytotoxicity in the high AFP-producing (HuH-7) but not in the low AFP-producing (huH-1/cl.2) human hepatoma cells. In the present study, we constructed the retroviral vector (LNAFM0.3TK) in which the HSVtk gene expression is regulated by the variant-type of the 0.3 kb human AFP promoter with a G-to-A substitution at nucleotide -119, a point mutation responsible for hereditary persistence of human AFP and the vector was applied to three human hepatoma cell lines HuH-7, huH 1/cl.2 and intermediate AFP-producing cells (PLC/PRF/5). By the reporter gene transfection assay, the activity of the variant-type of the promoter was much higher than that of the wild-type of the promoter in both HuH-7 and huH-1/cl.2 cells. Consistent with this, LNAFM0.3TK infection could sensitize huH-1/cl.2 cells, as well as HuH-7 and PLC/PRF/5 cells to GCV, but did not affect cell growth of nonhepatoma cells (HeLa). In addition, the bystander effect was achieved more efficiently by LNAFM0.3TK infection than LNAFW0.3TK infection in HuH-7 cells. These results suggest that the variant-type of the human AFP promoter ensures the therapeutic gene expression in gene therapy particularly for the low AFP-producing hepatoma cells. PMID- 10476207 TI - Variation in the immune response to adenoviral vectors in the brain: influence of mouse strain, environmental conditions and priming. AB - E1-deleted adenoviral vectors expressing the bacterial beta-galactosidase gene were inoculated into the brain of unprimed and primed C3H.He or C57BL/6J mice housed under either conventional or specific-pathogen-free (SPF) conditions. The kinetics of immune responses to both the vector and the transgene were investigated. In mice previously sensitized to adenovirus, the leukocyte infiltrate in the brain was dominated by CD8+ T cells, whereas in unprimed mice CD4+ T cells were present at higher levels. As expected, antibody titres to both adenovirus and beta-galactosidase were higher in primed mice than in unprimed mice after intracranial inoculation. C3H.He mice consistently made higher antibody responses than C57BL/6J mice. Although adenoviral vectors induced an inflammatory response under all conditions, mice housed in SPF facilities exhibited less inflammation than conventional mice and transgene expression persisted for longer. Irrespective of whether the mice had been deliberately primed to adenovirus, antibody titres were consistently lower in SPF mice compared with conventional mice. This study clearly demonstrates that environmental conditions, as well as previous priming to adenovirus, will affect both the quality and duration of the immune response triggered by gene delivery to the brain. PMID- 10476208 TI - Metabolic instability of plasmid DNA in the cytosol: a potential barrier to gene transfer. AB - Inefficient nuclear delivery of plasmid DNA is thought to be one of the daunting hurdles to gene transfer, utilizing a nonviral delivery system such as polycation DNA complex. Following its internalization by endocytosis, plasmid DNA has to be released into the cytosol before its nuclear entry can occur. However, the stability of plasmid DNA in the cytoplasm, that may play a determinant role in the transfection efficiency, is not known. The turnover of plasmid DNA, delivered by microinjection into the cytosol, was determined by fluorescence in situ hybridization (FISH) and quantitative single-cell fluorescence video-image analysis. Both single- and double-stranded circular plasmid DNA disappeared with an apparent half-life of 50-90 min from the cytoplasm of HeLa and COS cells, while the amount of co-injected dextran (MW 70,000) remained unaltered. We propose that cytosolic nuclease(s) are responsible for the rapid-degradation of plasmid DNA, since (1) elimination of plasmid DNA cannot be attributed to cell division or to the activity of apoptotic and lysosomal nucleases; (2) disposal of microinjected plasmid DNA was inhibited in cytosol-depleted cells or following the encapsulation of DNA in phospholipid vesicles; (3) generation and subsequent elimination of free 3'-OH ends could be detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay (TUNEL), reflecting the fragmentation of the injected DNA; and finally (4) isolated cytosol, obtained by selective permeabilization of the plasma membrane, exhibits divalent cation dependent, thermolabile nuclease activity, determined by Southern blotting and 32P-release from end-labeled DNA. Collectively, these findings suggest that the metabolic instability of plasmid DNA, caused by cytosolic nuclease, may constitute a previously unrecognized impediment for DNA translocation into the nucleus and a possible target to enhance the efficiency of gene delivery. PMID- 10476209 TI - Murine MHC class II locus control region drives expression of human beta glucocerebrosidase in antigen presenting cells of transgenic mice. AB - Gaucher disease is the most prevalent lysosomal storage disorder in humans, resulting from an inherited deficiency of the enzyme glucocerebrosidase. Although the enzyme is ubiquitously expressed, cells of the reticuloendothelial system are particularly affected since they accumulate the undigested glucosylceramide substrate through their role in scavenging and breaking down cell debris. Gaucher disease is an attractive target for somatic gene therapy. To test the ability to express the enzyme in the affected cell types we have generated transgenic mice expressing human glucocerebrosidase under the control of the murine major histocompatibility complex (MHC) class II Ead locus control region (LCR). The four transgenic lines express the human enzyme in a copy number-dependent manner, independent of the integration site of the transgene. Over-expression of the human enzyme in mice did not result in any abnormal phenotype or pathology during the period of observation (> 2 years). The enzyme is expressed in B cells, monocytes, dendritic cells, thymic epithelial cells, and macrophages in various tissues: the peritoneal cavity, bone marrow, spleen, kidney, gastrointestinal tract, Kupffer cells in the liver and alveolar macrophages in lungs. Expression in the brain was limited to perivascular macrophages and was not seen in microglial cells. Therefore, the MHC class II LCR could potentially be of use in somatic gene therapy for type 1 Gaucher disease. PMID- 10476210 TI - Electropermeabilization of skeletal muscle enhances gene transfer in vivo. AB - This work demonstrates that electrical muscle stimulation markedly increases the transfection efficiency of an intramuscular injection of plasmid DNA. In soleus or extensor digitorum longus muscles of adult rats the percentage of transfected fibers increased from about 1 to more than 10. The number of transfected fibers and the amount of foreign protein produced could be graded by varying the number or duration of the electrical pulses applied to the muscle. The stimulation had to be applied when DNA was present in the muscle. When dextran was injected together with the plasmid DNA, it was also taken up by the transfected fibers. Stimulation-induced membrane permeabilization and increased DNA uptake were therefore probably responsible for the improved transfection. The stimulation caused some muscle damage but the fibers regenerated rapidly. The described method, which is simple, efficient, and reproducible, should become valuable for basic research, gene therapy and DNA vaccination. PMID- 10476211 TI - Adenoviral reporter gene transfer to the human trabecular meshwork does not alter aqueous humor outflow. Relevance for potential gene therapy of glaucoma. AB - Obstruction of the aqueous humor outflow from the anterior chamber of the eye leads to an elevation of intraocular pressure in glaucoma, the second major cause of blindness worldwide. Our goal is to be able to modulate aqueous humor outflow resistance by gene transfer to the cells of the trabecular meshwork (TM). We have previously shown that adenoviral vectors are able to transfer a reporter gene to the TM of postmortem human donors. However, assessing gene therapy for glaucoma requires models that can monitor changes in aqueous humor outflow facility (C = flow/pressure). In this study we used four replication-deficient adenoviruses in two such perfusion models. In the first model, whole porcine eyes were infected, perfused at constant pressure and flow changes recorded for 5 h. In the second one, anterior segments from human eyes were infected, perfused at constant flow and pressure changes recorded for 3 days. A single dose of 10(8) adenovirus plaque forming units (pfu) causes a reduction in C while single doses of 10(7), 10(6) and 10(5) p.f.u. do not affect outflow facility and retain positive gene transfer. These findings indicate that adenovirus, at effective doses, could become useful vectors for gene therapy of glaucoma. PMID- 10476212 TI - Complete cure of established murine hepatocellular carcinoma is achievable by repeated injections of retroviruses carrying the herpes simplex virus thymidine kinase gene. AB - Although xenotransplantation of retrovirus-producing cells into a tumor has been shown to be effective for the treatment of cancer, injections of recombinant retroviruses are much more feasible for clinical applications. We established a clone producing retroviruses carrying the herpes simplex virus thymidine kinase (HSVtk) gene with titers of up to 4 x 10(7) colony-forming units/ml, and examined the effectiveness of in vivo gene therapy against cancer. Syngeneic mice were inoculated subcutaneously with murine hepatocellular carcinoma (HCC) cells, BNL1ME A.7R.1, and the treatment was initiated after tumors were established. When mice were given an intratumoral injection of HSVtk-carrying retroviruses or their producing cells followed by ganciclovir (GCV) treatment, significantly prolonged survival periods were observed. When mice were treated with repeated intratumoral injections of HSVtk-carrying retrovirus-producing cells, significant antitumor responses and some cures were induced by GCV treatment. Furthermore, repeated intratumoral injections of HSVtk-carrying retroviruses and GCV treatment resulted in complete regression of established HCC tumors in all animals used in the experiment. Mice that completely eradicated tumors exhibited protective immunity against wild-type HCC tumors. These results suggest that repeated injections of HSVtk-carrying retroviruses followed by GCV treatment is a potent modality for the treatment of solid tumors. PMID- 10476213 TI - The extra- and intracellular barriers to lipid and adenovirus-mediated pulmonary gene transfer in native sheep airway epithelium. AB - Gene transfer to the respiratory epithelium is currently suboptimal and may be helped by the identification of limiting biological barriers. We have, therefore, developed an ex vivo model which retains many of the characteristics of in vivo native airways including mucociliary clearance, mucus coverage and an intact cellular structure. Using this model we have demonstrated several barriers to gene transfer. Liposome-mediated gene transfer was inhibited by normal mucus, with removal of this layer increasing expression approximately 25-fold. In addition both liposome and adenovirus were inhibited by CF sputum. The apical membrane represented a significant barrier to both agents. Adenovirus-mediated expression could be significantly augmented by increasing contact time or by pre treatment of tissues with a nominally calcium-free medium. The presence of these extracellular and plasma membrane barriers appeared to be the key parameters responsible for the approximately three log difference in gene expression found in vitro compared with our ex vivo model. Cytoskeletal elements and the cell cycle also influenced in vitro gene transfer, and represent further barriers which need to be overcome. PMID- 10476214 TI - Enhanced pancreatic tumor regression by a combination of adenovirus and retrovirus-mediated delivery of the herpes simplex virus thymidine kinase gene. AB - We have evaluated the effectiveness of combining the different characteristics of retrovirus and adenovirus to apply the herpes simplex virus thymidine kinase gene (HSVtk) and ganciclovir (GCV) treatment for gene therapy of pancreatic cancer. Transduction of NP-18 human pancreatic cells in culture by either the adenoviral vector (ADV/tk) or the retroviral vector (Rv/tk) followed by GCV treatment resulted in a GCV dose-dependent cytotoxic effect. A bystander effect was determined, both in NP-18 cultures and in xenogeneic cell mixtures of NP-18 and PA317 cells. Studies in vivo indicated that the effectiveness of tumor regression after HSVtk gene transfer and GCV treatment was dependent first on the tumor size at the time of viral injection and secondly, in large tumors, on the type of virus administered. The administration of the viral combination (ADV/tk + vector producer cells VPC-Rv/tk) was the best approach tested and resulted in a dramatic reduction in tumor mass after 4 days of GCV treatment which was maintained for the treatment period. Remarkably, two animals presented a complete eradication of the tumor. Thus, the HSVtk/GCV system when administered using a viral combination (ADV/tk + VPC-Rv/tk), may be a promising suicide gene therapy for pancreatic carcinomas. PMID- 10476215 TI - Adenoviral delivery of CTLA4Ig into myeloid dendritic cells promotes their in vitro tolerogenicity and survival in allogeneic recipients. AB - Dendritic cells (DC) are highly specialized antigen-presenting cells (APC) that initiate and modulate immune responses. They are essential for naive T cell activation, but may also play roles both in central and peripheral tolerance. Blockade of costimulatory pathways that provide the crucial second signal for lymphocyte activation is one strategy to augment the potential tolerogenicity of DC. Here, in vitro propagated DC were transduced using an adenoviral (Ad) vector to express the gene encoding cytotoxic T lymphocyte antigen 4-immunoglobulin (CTLA4lg), which blocks interaction of CD80 and CD86 on DC with CD28 on T cells. Supernatants of AdCTLA4lg-transduced DC strikingly inhibited mixed leukocyte reactions (MLR) induced by non-transduced DC. Whereas transduction of marker genes (LacZ or enhanced green fluorescence protein (EGFP)) did not alter their potent allostimulatory activity, DC transduced with CTLA4lg exhibited striking reductions in cell surface staining for CD86, but not MHC class II, and were poor stimulators of T cell proliferation and cytotoxic T lymphocyte (CTL) responses. In addition, they induced alloantigen-specific T cell hyporesponsiveness. They were detected, following local injection, in significantly increased numbers in the lymphoid tissue of unmodified allogeneic recipients. This is the first report of the functional properties of DC genetically engineered to express CTLA4lg. PMID- 10476216 TI - Hepatoma-specific antitumor activity of an albumin enhancer/promoter regulated herpes simplex virus in vivo. AB - Targeting viral vectors to appropriate cell types so that normal cells are not adversely affected is an important goal for gene therapy. Previously, we described a novel approach to viral gene therapy using a conditional, replication competent herpes simplex virus (HSV), where replication and associated cytotoxicity are limited to a specific cell-type by the regulated expression of an essential immediate-early viral gene product. In this report we analyze the hepatoma-specific replication, cytotoxicity and anti-tumor effect of recombinant HSV G92A, regulated by the albumin enhancer/promoter. G92A efficiently replicated in vitro in two human hepatoma cell lines expressing albumin, but not in four human non-hepatoma, albumin-non-expressing tumor cell lines, while all cell lines were equally susceptible to a tissue nonspecific HSV recombinant, hrR3. In vivo, G92A replicated well in subcutaneous xenografts of human hepatoma cells (Hep3B) in athymic mice, but not in non-hepatoma subcutaneous tumors (PC3 and HeLa), whereas, hrR3 replicated well in both tumor types. Intratumoral inoculation of G92A inhibited the growth of established subcutaneous hepatoma tumors in nude mice, but not prostate tumors. Replication-competent viral vectors controlled by cell-specific transcriptional regulatory sequences provide a new therapeutic strategy for tumor therapy. PMID- 10476217 TI - Macrophage inflammatory protein-2 gene therapy attenuates adenovirus- and acetaminophen-mediated hepatic injury. AB - Profound hepatocellular injury is often a consequence of adenovirus-mediated gene therapy or acetaminophen ingestion. The aim of the present study was to examine the role of a CXC chemokine, macrophage inflammatory protein-2 (MIP-2), in the hepatotoxic response by mice infected with adenovirus and challenged with acetaminophen. CD1 mice that received a replication-defective human type 5 adenovirus vector (Ad70-3) intravenously exhibited hepatic injury that peaked at 24 h after infection. In contrast, mice that received a similar adenovirus vector containing a rodent MIP-2 cDNA insert had no hepatic injury at any time after infection. The combination of Ad70-3 infection and an intraperitoneal challenge with 400 mg/kg of acetaminophen was fatal in 50% of the mice, but only 10% of the AdMIP-2 group receiving acetaminophen were similarly affected. Furthermore, AdMIP 2 mice had significantly lower hepatic injury and serum aminotransaminases compared with the Ad70-3 group. However, AdMIP-2 infection in mice lacking the CXC chemokine receptor that binds MIP-2, CXCR2, did not attenuate any of the markers of liver injury after adenovirus and acetaminophen challenge. AdMIP-2 treatment of CD1 mice was also associated with significantly decreased leukocyte infiltration into the liver and an earlier increase in hepatic 3H-thymidine incorporation compared with the control group. Taken together, these data demonstrate that MIP-2 has a protective role in both adenovirus- and acetaminophen-mediated hepatotoxicity, and suggest that MIP-2 may promote rapid hepatic regeneration following acute hepatic injury. PMID- 10476218 TI - Dynamic changes in the characteristics of cationic lipidic vectors after exposure to mouse serum: implications for intravenous lipofection. AB - Intravenous gene delivery via cationic lipidic vectors gives systemic gene expression particularly in the lung. In order to understand the mechanism of intravenous lipofection, a systematic study was performed to investigate the interactions of lipidic vectors with mouse serum emphasizing how serum affects the biophysical and biological properties of vectors of different lipid compositions. Results from this study showed that lipidic vectors underwent dynamic changes in their characteristics after exposure to serum. Addition of lipidic vectors into serum resulted in an immediate aggregation of vectors. Prolonged incubation of lipidic vectors with serum led to vector disintegration as shown in turbidity study, sucrose-gradient centrifugation analysis and fluorescence resonance energy transfer (FRET) study. Vector disintegration was associated with DNA release and degradation as shown in EtBr intercalation assay and DNA digestion study. Serum-induced disintegration of vectors is a general phenomenon for all cationic lipidic vectors tested in this study. Yet, vectors of different lipid compositions vary greatly in the rate of disintegration. There is an inverse correlation between the disintegration rate of lipidic vectors and their in vivo transfection efficiency. Vectors with a rapid rate of disintegration such as those containing dioleoyl-phosphatidylethanolamine (DOPE) poorly stayed in the lung and were barely active in transfecting cells. In contrast, cholesterol-containing vectors that had a rapid aggregation and a slow disintegration were highly efficient in transfecting cells in vivo. The results of this study explain why cationic lipidic vectors of different lipid compositions have a dramatic difference in their in vivo transfection efficiency. These results also suggest that the study of the interactions of lipidic vectors with serum may serve as a predictive model for the in vivo efficiency of a lipidic vector. Further study of the numerous interactions of lipidic vectors with serum might lead to the development of a vector which can deliver a gene to target cells in a tissue-specific manner. PMID- 10476219 TI - PEGylated DNA/transferrin-PEI complexes: reduced interaction with blood components, extended circulation in blood and potential for systemic gene delivery. AB - We investigated the in vitro and in vivo properties of DNA/transferrin polyethylenimine (800 kDa) complexes before and after covalent coupling of poly(ethylene glycol) (PEG). Upon incubation with plasma, the positively charged non-PEGylated DNA complexes form aggregates. Plasma proteins such as IgM, fibrinogen, fibronectin and complement C3 were found to bind to non-PEGylated DNA complexes. At DNA concentrations relevant for in vivo gene delivery a strong aggregation of erythrocytes was also observed. PEGylation of the complexes strongly reduces plasma protein binding and erythrocyte aggregation. Furthermore, PEGylated complex size was stabilized and had a reduced surface charge. Prolonged circulation in the blood of the PEGylated complexes was also observed when injected intravenously. In tumor bearing mice, application of non-PEGylated complexes through the tail vein resulted in reporter gene expression in tail and lung, but severe toxicity was observed in some mice. In contrast, PEGylated complexes mediated reporter gene transfer to the tumor without significant toxicity. PMID- 10476220 TI - Transfer of the murine interleukin-12 gene in vivo by a Semliki Forest virus vector induces B16 tumor regression through inhibition of tumor blood vessel formation monitored by Doppler ultrasonography. AB - To elucidate further the potential of a Semliki Forest virus (SFV) vector in vivo for gene therapy, we constructed a vector, SFV-IL12, to transfer murine IL-12 genes into tumors. A single intratumoral injection of established B16 murine melanoma with SFV-IL12 resulted in a significant inhibition of tumor growth, while injection with SFV-LacZ had no effect. This antitumoral activity correlated with an increase of IFN gamma production, MIG and IP-10 mRNA expression, both at the tumor site and at the periphery. In contrast, no increase in CTL- or NK cell mediated cytotoxic response could be detected, ruling out the involvement of T and NK cell cytotoxicity. To determine how the transfer to IL-12 genes induced tumor regression, the antiangiogenic-activity of SFV-IL12 was investigated using Doppler ultrasonography (DUS). SFV-IL12 inhibited in situ neovascularization within the tumor, without affecting the resistance index of pre-existing intratumoral blood flows. In addition, histological analysis of SFV-IL12-treated tumors showed massive tumor necrosis induced by SFV-IL12 treatment. These data indicate that SFV-IL12 inhibits tumor growth through its antiangiogenic activity, demonstrated for the first time in vivo by DUS, and suggest that the SFV vector may be a novel valuable tool in tumor gene transfer. PMID- 10476221 TI - Design of a muscle cell-specific expression vector utilising human vascular smooth muscle alpha-actin regulatory elements. AB - The facility to direct tissue-specific expression of therapeutic gene constructs is desirable for many gene therapy applications. We describe the creation of a muscle-selective expression vector which supports transcription in vascular smooth muscle, cardiac muscle and skeletal muscle, while it is essentially silent in other cell types such as endothelial cells, hepatocytes and fibroblasts. Specific transcriptional regulatory elements have been identified in the human vascular smooth muscle cell (VSMC) alpha-actin gene, and used to create an expression vector which directs the expression of genes in cis to muscle cells. The vector contains an enhancer element we have identified in the 5' flanking region of the human VSMC alpha-actin gene involved in mediating VSMC expression. Heterologous pairing experiments have shown that the enhancer does not interact with the basal transcription complex recruited at the minimal SV40 early promoter. Such a vector has direct application in the modulation of VSMC proliferation associated with intimal hyperplasia/restenosis. PMID- 10476222 TI - Co-delivery of T helper 1-biasing cytokine genes enhances the efficacy of gene gun immunization of mice: studies with the model tumor antigen beta-galactosidase and the BALB/c Meth A p53 tumor-specific antigen. AB - DNA-based immunization is currently being investigated as a new method for the induction of cellular and humoral immunity directed against viral disease and cancer. In the present study we characterized and compared the immune responses induced in mice following particle-bombardment of the skin ('gene gun' immunization) with those elicited by intracutaneous injection of a recombinant adenoviral vector. Using the well characterized beta-galactosidase (beta gal) model Ag system we find that both in vivo gene transfer systems elicit potent and long-lasting anti-beta gal-specific CD8+ and CD4+ T cell responses. However, gene gun immunization predominantly promotes the production of anti-beta gal antibodies of the gamma 1 isotype, indicative of a Th2-biased immune response, while intradermal injection of recombinant adenovirus primarily leads to the production of anti-beta gal gamma 2a antibodies, indicative of a Th1-biased immune response. Since viral infections are generally associated with the production of large amounts of IFN-alpha and IL-12, we investigated whether administration of expression plasmids encoding these Th1-associated cytokines along with antigen-encoding cDNA can influence the nature of the immune response resulting from gene gun immunization. We observed that co-delivery of IFN-alpha or IL-12 resulted in increased production of anti-beta gal gamma 2a antibodies. This suggests a shift towards a Th1 phenotype of the resulting immune response, thus mimicking a viral infection. Importantly, gene gun immunization of mice with a naturally occurring tumor antigen, the tumor-specific p53 mutant antigen expressed by the chemically induced BALB/c Meth A sarcoma, required co-delivery of IL-12 for the induction of effective antitumor immunity. These results have important implications for the design of clinically relevant gene gun immunization strategies for tumor immunotherapy. PMID- 10476223 TI - Inhibition of adenovirus-mediated gene transfer by bronchoalveolar lavage fluid. AB - Human and animal trials with recombinant adenovirus have been discouraging, since the level of recombinant gene expression was low. Nonspecific and specific immune response mediated by, for example, macrophages, T cells and immunoglobulins may prevent infection or cause death of infected cells. We analyzed the effect of bronchoalveolar lavage fluid (BAL) on the efficiency of adenoviral infection in vitro. A total of 26 BAL samples of randomly selected patients was examined. Adenovirus-mediated gene transfer was quantified using AdCMV. Null, a recombinant adenovirus, in a modified titer assay based on immunocytochemical detection of infected 293 cells. In addition, the concentration of anti-adenovirus-type 5 IgA, IgG and IgM antibodies in BAL was determined by ELISA. 53.8% of the BAL samples (14 out of 26) reduced adenoviral infectivity by at least 50% (factor of inhibition > or = 2). All BAL samples effecting, a reduction in adenoviral infectivity contained detectable amounts of anti-adenovirus-type 5-IgA antibodies. However, the correlation between the concentration of IgA antibody and the strength of inhibition was weak (r = 0.336). Even high levels of anti adenovirus-type 5-IgM, IgG or IgA antibodies did not influence adenoviral infectivity consistently. This observation indicates that BAL contains (a) anti adenovirus-type 5 antibodies which are not directed against adenoviral epitopes responsible for the viral adherence and uptake process; and/or (b) inhibitors of viral infectivity different from antibodies. PMID- 10476224 TI - Factors affecting blood clearance and in vivo distribution of polyelectrolyte complexes for gene delivery. AB - Self-assembling polycation/DNA complexes represent a promising synthetic vector for gene delivery. However, despite considerable versatility and transfectional activity in vitro, such materials are quickly eliminated from the bloodstream following intravenous injection (plasma alpha half-life typically less than 5 min). For targeted systemic delivery a more prolonged plasma circulation of the vector is essential. Here we have examined factors contributing to rapid elimination of poly(L-lysine) (pLL)/DNA complexes from the bloodstream, and implicate the binding of proteins to the polyelectrolyte complexes as a likely cause for their blood clearance. pLL/DNA complexes reisolated from serum associate with several proteins, depending on their net charge, although the major band on SDS-PAGE co-migrates with albumin. Serum albumin binds to pLL/DNA complexes in vitro, forming a ternary pLL/DNA/albumin complex which regains some ethidium bromide fluorescence and fails to move during agarose electrophoresis. Albumin also causes increased turbidity of complexes, and reduces their zeta potential to the same level (-16 mV) as is measured in serum. We propose that rapid plasma elimination of polycation/DNA complexes results from their binding serum albumin and other proteins, perhaps due to aggregation and phagocytic capture or accumulation of the ternary complexes in fine capillary beds. PMID- 10476225 TI - Lipoplex size is a major determinant of in vitro lipofection efficiency. AB - The inhibition effect of serum on the transfection efficiency of cationic liposome-DNA complexes (lipoplexes) is a major obstacle to the application of this gene delivery vector both in vitro and in vivo. The size of the lipoplexes, as they are presented to targeted cells, is found to be the major determinant of their effectiveness in transfection. The transfection efficiency and the cell association and uptake of lipoplexes with CHO cells was found to increase with increasing lipoplex size. The influence on the transfection efficiency of lipoplexes by their cationic lipid:DNA ratios, types of liposomes, incubation time in polyanion containing media, and time of serum addition, are mediated mainly through size. Lipoplexes at a 2:1 charge ratio grow in size in media containing polyanions. The size growth may be arrested by adding serum to the incubation media. By using large lipoplexes, especially those made from multilamellar vesicles, the serum inhibition effect may be overcome. PMID- 10476226 TI - Inhibition of HIV-1 by an anti-integrase single-chain variable fragment (SFv): delivery by SV40 provides durable protection against HIV-1 and does not require selection. AB - Human immunodeficiency virus type 1 (HIV-1) encodes several proteins that are packaged into virus particles. Integrase (IN) is an essential retroviral enzyme, which has been a target for developing agents to inhibit virus replication. In previous studies, we showed that intracellular expression of single-chain variable antibody fragments (SFvs) that bind IN, delivered via retroviral expression vectors, provided resistance to productive HIV-1 infection in T lymphocytic cells. In the current studies, we evaluated simian-virus 40 (SV40) as a delivery vehicle for anti-IN therapy of HIV-1 infection. Prior work suggested that delivery using SV40 might provide a high enough level of transduction that selection of transduced cells might be unnecessary. In these studies, an SV40 expression vector was developed to deliver SFv-IN (SV(Aw)). Expression of the SFv IN was confirmed by Western blotting and immunofluorescence staining, which showed that > 90% of SupT1 T-lymphocytic cells treated with SV(Aw) expressed the SFv-IN protein without selection. When challenged, HIV-1 replication, as measured by HIV-1 p24 antigen expression and syncytium formation, was potently inhibited in cells expressing SV40-delivered SFv-IN. Levels of inhibition of HIV-1 infection achieved using this approach were comparable to those achieved using murine leukemia virus (MLV) as a transduction vector, the major difference being that transduction using SV40 did not require selection in culture whereas transduction with MLV did require selection. Therefore, the SV40 vector as gene delivery system represents a novel therapeutic strategy for gene therapy to target HIV-1 proteins and interfere with HIV-1 replication. PMID- 10476227 TI - Efficient CFTR expression from AAV vectors packaged with promoters--the second generation. AB - Gene therapy studies of cystic fibrosis (CF) have shown that AAV-based vector was efficient in transferring but not in expressing the CFTR cDNA in the target cells. The levels of CFTR gene expression were limited by the small packaging capacity of AAV because it had been difficult to package the CFTR cDNA with an efficient promoter. In the present study we have developed a new generation of AAV/CFTR vectors which contain efficient short promoters to express the CFTR gene in target cells. To do so, we reduced the size of the CFTR cDNA by determining the minimal untranslated regions required for expression of CFTR cDNA. We also identified short and efficient promoters that could be packaged with the down sized CFTR cDNA into a novel AAV vector that had a maximal packaging capacity. Functional analyses showed that the new vectors were packaged efficiently and expressed higher levels of CFTR than a vector in which the CFTR gene was driven by the ITR sequence of AAV. Transduction of airway epithelial cells containing [symbol: see text] 508 mutation with the new vectors demonstrated efficient expression of the wild-type CFTR and correction of the CF phenotype. In contrast, no significant CFTR expression was detected in cells infected with the vector that express the CFTR gene from the ITR. These findings support the notion that the AAV can be developed into an efficient vector to transduce the CFTR gene and vectors expressing higher levels of CFTR from an efficient promoter should provide better efficacy for gene therapy of cystic fibrosis. PMID- 10476228 TI - Enhancement of calcium phosphate-mediated transfection by inclusion of adenovirus in coprecipitates. AB - Although coprecipitates of plasmid DNA and calcium phosphate (DNA:CaPi coprecipitates) were one of the first methods used for transfection of mammalian cells, they are inefficient for many cell types. Based on the recent finding that incorporating recombinant adenovirus in CaPi coprecipitates enhances expression of virus-encoded transgenes, we tested the hypothesis that including adenovirus in DNA:CaPi coprecipitates would increase the efficiency of transfection. We found that including adenovirus at the time of coprecipitate formation increased transgene expression in several cell types. Only a short incubation with cells was required, and the coprecipitates could be delivered in the presence of serum. Inclusion of adenovirus in coprecipitates did not increase DNA uptake by cells, and inactivated virus was also effective. Neutralizing anti-hexon antibody attenuated the enhancement produced by incorporating virus. These data suggest that the virus enhanced expression at a step after cellular uptake, probably by increasing DNA release from endosomes. The DNA:CaPi:Ad coprecipitates were at least as effective as complexes of DNA with adenovirus and polethylenimine or Lipofectin, but produced less cellular toxicity. The results suggest that DNA:CaPi:Ad coprecipitates have advantages for in vitro gene transfer and provide an attractive vehicle for investigating cellular mechanisms of gene transfer. PMID- 10476229 TI - Gene delivery to the myocardium by intrapericardial injection. AB - Several studies have demonstrated the feasibility of gene transfer into the heart muscle. However, all the available data also indicate that the extent of transfection remains limited. As an alternative method to intravascular administration, we have developed a novel strategy which uses the pericardial sac. When a replication-deficient adenovirus containing the cDNA encoding a bacterial beta-galactosidase is injected into the pericardial sac of adult Wistar rats the staining is exclusively restricted to the pericardial cell layers. However, injecting a mixture of collagenase and hyaluronidase together with the virus, leads to a large diffusion of the transgene activity, reaching up to 40% of the myocardium. Transgene expression is predominant in the left ventricle and the interventricular septum but limited in the right ventricle. In vivo echocardiographic measurements of the left ventricular diameters at end diastolic and end systolic times show no difference between virus- and sham-injected animals, thus indicating a good clinical tolerance to this strategy of virus delivery. The same protocol has been used with the same efficiency in mice, which leads us to propose injection into the pericardial sac as an effective and harmless method for gene transfer into the heart muscle. PMID- 10476230 TI - In vivo targeted gene transfer into liver cells mediated by a novel galactosyl-D lysine/D-serine copolymer. AB - A novel synthetic polypeptide designed as a DNA binding-molecule for liver specific, receptor-mediated, gene transfer was used to selectively introduce reporter genes into liver cells in the form of plasmid DNA-ligand complexes. The polypeptide was a D-lysine/D-serine copolymer (Lys/Ser = 33/36 or 53/60) modified with a polyethylene glycol 5000 at the carboxyl-terminus (PLSP). In addition, the lysine epsilon-amino groups were covalently bound to galactose (galactosyl-PLSP), a ligand for the asialoglycoprotein receptor on hepatocytes. Solutions of DNA ligand complex were prepared by adding solutions of DNA and galactosyl-PLSP in a mixing ratio of DNA:galactosyl-PLSP = 1:3 (w/w). Following injection of the DNA ligand complex into mice via the tail vein, high levels of luciferase and enhanced green fluorescent protein, which were encoded by the reporter genes, were observed in liver. In contrast, luciferase activity in kidney, spleen, lung and heart was negligible. The high levels of gene expression obtained with DNA/galactosyl-PLSP complexes were achieved without partial hepatectomy or administration of lysosomotrophic agents. Thus, the synthetic Lys/Ser copolymer used in the present study appears to be a promising new tool which enhances the efficacy of receptor-mediated gene transfer into hepatocytes and which may provide another step toward the clinical practice of organ-specific gene therapy. PMID- 10476231 TI - Antigen-specific membrane fusion mediated by the haemagglutinin protein of influenza A virus: separation of attachment and fusion functions on different molecules. AB - Using genetic engineering techniques, two strategies for changing the receptor binding specificity of the influenza A virus haemagglutinin (HA) protein whilst preserving its membrane fusion function, have been explored. The aim was to investigate whether the HA protein could be developed as an attachment/entry protein for targeting enveloped virus gene therapy vectors to specific cell populations. In the first strategy, a single chain antibody Fv region (scFv) specific for the hapten NIP was inserted between HA1 codons 139 and 145, to create a scFv-HA chimaeric protein. This protein was shown to possess anti-NIP binding activity, but membrane fusion activity could not be demonstrated. The possibility that linking the scFv domain directly to HA may have inhibited the HA fusion function led to the development of the second strategy. This involved separating the receptor-binding and membrane fusion functions of HA on to two different molecules. The feasibility of this strategy was tested by looking for fusion between NIP-conjugated red blood cells which lacked sialic acid (the HA protein's natural receptor) and Chinese hamster ovary cells that expressed both the above anti-NIP scFv-HA chimaeric protein (as a non-fusigenic, receptor binding molecule) and wild-type HA protein (as a fusigenic, non-binding molecule) on their surface. Cell-to-cell fusion was observed in this system, indicating that the receptor-binding function of HA can be transferred to an adjacent molecule, and also changed in its specificity, without compromising its membrane fusion activity. This finding strongly suggests that the development of a two molecule attachment and entry system for retargeting enveloped virus gene therapy vectors, based on HA, is a viable proposition. PMID- 10476232 TI - 5-Azacytidine prevents transgene methylation in vivo. AB - Retroviral sequence can silence transgene expression in vitro and in vivo. We report that this effect can be efficiently prevented by in vivo administration of the demethylating agent 5-azacytidine (aza-C). We engineered the U937 human cell line with a retroviral vector consisting of the thymidine kinase suicide gene (tk), which induces sensitivity to ganciclovir (gcv) and through an IRES sequence, the bacterial beta-galactosidase gene (lacZ) as a marker gene. About 90% of the U937 cells expressed the transgene. By injecting the transduced U937 cells in severe combined immunodeficient disease (SCID) mice, we generated a tumor which, during in vivo treatment with aza-C, maintained the high expression of lacZ and tk genes at the baseline values. LacZ-positive cells in the tumour masses after death was weak (1-2%) in the control group, while in mice treated with aza-C it was maintained at 90%. The delay in tumour onset was significantly longer when animals were treated with both aza-C and gcv (P < 0.0001) compared with animals treated with gcv or with aza-C alone. The prevention of silencing phenomena has important implications for gene therapy, because an efficient transduction associated with appropriate drug therapy, might be a powerful strategy for successful application of gene therapy protocols. PMID- 10476233 TI - Maternal mortality: lessons from the confidential enquiry. PMID- 10476234 TI - Sildenafil: desired and undesired effects. PMID- 10476235 TI - Methadone: where are we now? AB - Methadone is now widely used as opioid replacement therapy because it reduces crime and prevents spread of disease by the intravenous route. However, it is potentially toxic, especially to drug naive patients, and requires careful prescribing. PMID- 10476236 TI - Alternative ways of using and abusing drugs and complicity of doctors. AB - Many drugs that have been prescribed by doctors are not taken as directed. Altered dose, frequency and route of administration may lead to misuse of drugs that have been prescribed in good faith. Awareness of the abuse potential of prescribed and over-the-counter drugs is the first point of intervention, followed by monitoring of each doctor's practice. PMID- 10476237 TI - Psychiatric aspects of alcohol misuse. AB - Alcohol and psychiatric disorder have a complicated relationship. Certain important psychiatric syndromes arise from the toxic effects of alcohol and thiamine deficiency. Liver disease and hypoglycaemia are also associated with their own psychiatric syndromes. Many psychiatric patients also turn to alcohol as an easily available 'medication'. PMID- 10476238 TI - Treatment of psychiatric aspects of alcohol misuse. AB - Psychiatric disorders and alcohol misuse often coexist. This article outlines the clinical management of these comorbid disorders. It discusses and suggests therapeutic recommendations. PMID- 10476239 TI - Techniques in conduit harvesting. AB - Coronary artery bypass grafting is a commonly performed operation for the treatment of ischaemic heart disease. The success of it is largely dependent upon the quality and type of graft used. This article describes the application of and rationale behind techniques commonly used in conduit harvesting. PMID- 10476240 TI - Nuclear cardiology in clinical practice. AB - This article is an overview of nuclear cardiology for the busy clinician. We discuss the use of myocardial perfusion scans, radionuclide ventriculography, positron emission tomography scans and the interpretation of nuclear scan reports. PMID- 10476242 TI - Interferon beta-1a in relapsing-remitting multiple sclerosis. AB - The accumulated results from multiple trials have unequivocally demonstrated beneficial effects of interferon beta-1a on disease activity in relapsing remitting multiple sclerosis, both for clinical and magnetic resonance endpoints. New data suggest that low doses administered once weekly are relatively ineffective and that higher, more frequent doses are required for optimal benefit. PMID- 10476241 TI - High intensity training and the heart. AB - Absence may make the heart grow fonder, but exercise makes the heart grow stronger. This review discusses the cardiac impact of high intensity training, and discusses the possible mechanisms underlying the risk and benefit of such training. PMID- 10476243 TI - Surgery for voice. AB - Preoperative assessment in a multidisciplinary voice clinic, using stroboscopy, is essential. In the treatment of benign vocal fold mucosal disorders precise phonosurgical techniques, using high magnification which respects the multilayered structured of the vocal fold, are mandatory. A paralysed vocal fold can be assessed by electromyography and medialization thyroplasty procedures offer excellent 'fine tuning' of the voice. PMID- 10476244 TI - Professional competence in medicine. AB - Professional competence in medicine is under the microscope following a year of government and media attention directed at the performance of doctors and outcomes for patients. The ability of the profession to self-regulate has been questioned and the roles of the state, the universities, the Royal colleges and the postgraduate deans are shifting. This paper provides a context for considering these important changes. PMID- 10476245 TI - Has surgical subspecialization gone too far? AB - Subspecialization is currently fashionable, but with present hospital organization, the need to cover emergencies and the lack of convincing evidence for its superiority, the tide is on the ebb and the place of the general surgeon is secure. PMID- 10476246 TI - Surgical training for the next millennium. AB - The Calman reforms in training have meant that surgical training now consists of a 2-year basic training period followed by a 6-year period of higher surgical training. This article addresses the problems faced by the new Calman surgical trainees and proposes some new measures which could be introduced for surgical training in the next millennium. PMID- 10476247 TI - Improving senior house officer training in accident and emergency medicine: the benefits of a diploma. AB - Accident and emergency is a fascinating specialty providing exposure to a wide range of acute and chronic conditions from all branches of medicine. To ensure a minimal standard of training nationally we propose the introduction of a diploma for junior doctors. PMID- 10476248 TI - An unusual cause of obstructive sleep apnoea. PMID- 10476249 TI - Osteoporotic fracture complicating Guillain-Barre syndrome. PMID- 10476250 TI - Cervical spine infection presenting as angina. PMID- 10476251 TI - High- vs low-dose ACE inhibition in chronic heart failure. PMID- 10476252 TI - Evaluation of treatment of recurrent small-cell lung cancer. PMID- 10476253 TI - Confidential enquiry into maternal mortality. PMID- 10476254 TI - Confidential enquiry into maternal mortality. PMID- 10476255 TI - Physician-assisted suicide: the debate continues. PMID- 10476257 TI - A rare cause of tracheal obstruction unrelieved by tracheal intubation. PMID- 10476258 TI - Last chance for PEPs. PMID- 10476259 TI - Fixed nail plates versus sliding hip systems for the treatment of trochanteric femoral fractures: a meta analysis of 14 studies. AB - In order to compare accurately fixed nail plates with sliding implants 13 randomised, quasi-randomised or comparative studies, involving a total of 2,855 patients, which compared these implants, were systematically reviewed. In addition the results of a further 214 patients treated in a prospective audit are presented. Results showed an increased risk of cut-out (13% versus 4%), non-union (2% versus 0.5%), implant breakage (14% versus 0.7%) and re-operation (10% versus 4%) for fixed nail plates in comparison with the sliding implants. In addition patients treated with fixed nail plates had a higher mortality and the survivors were more likely to have residual pain in the hip and impaired mobility. The continued use of fixed nail plates cannot be justified given the unequivocal superiority of the sliding hip screw. PMID- 10476260 TI - The association between seniority of Accident and Emergency doctor and outcome following trauma. AB - The actual survival of patients treated following trauma in four Scottish Teaching hospitals during five years was compared with predicted survival according to TRISS analysis. The data were analysed according to the seniority of the Accident and Emergency (A&E) doctor treating each patient. The group of patients treated by a consultant had a significantly better outcome (p < 0.05) than the group of patients treated by junior doctors. Analysis of outcome according to the grade of junior A&E doctor suggested a step-wise improvement in outcome with seniority, thus supporting the concept that an improved outcome is associated with experience and seniority. These results support calls for A&E consultants to be increasingly involved in the management of patients with major trauma. Such increased involvement would require an increase in the number of A&E consultants. PMID- 10476261 TI - Should we operate on nonagenarians with hip fractures? AB - Studies on fractured necks of femurs usually have a cut off point at the age of 85. However, in view of an ageing populace it seems reasonable to look at the nature of hip fractures in the nonagenarians. In previous studies mortality and morbidity of 90-year-olds has been high. It has also been suggested that operating on the demented and institutionalised is not particularly effective. A 4-year review of 164 nonagenarian patients with hip fractures was therefore conducted at York district hospital. Access to the records of 136 (83%) patients was obtained with limited admission and mortality data from a further 28. Mortality rates at six months were 43% (24% in hospital) increasing to 46% at one year. Sixty-two patients had some complication of which 35 were dead within 6 months. Sixty-two per cent of patients returned to their pre admission residence. The mortality rates in this review are lower than in some previous studies and of those that survived 6 months 53% had no postoperative complications. It would therefore seem reasonable to operate on nonagenarians. PMID- 10476262 TI - Detection of pneumoperitoneum by ultrasound examination: an experimental and clinical study. AB - PURPOSE OF THE STUDY: The aim of the study is to show the value and reliability of ultrasound examination in detecting free intraabdominal air after blunt abdominal trauma. METHODS: The experimental part of this study includes the evaluation of the sonographic morphology of air within liquid and solid tissues. Subsequently air was injected into the abdominal cavities of 10 cadavers and sonography utilized for localization and assessment of the minimal detectable air volume. The clinical part of the study reports on the sonographic detection of hollow viscus perforation in 10 patients, who were admitted to our institution because of acute abdominal problems. RESULTS: Sonography is a reliable method for the diagnosis of free intraabdominal air. Best position for ultrasound examination of the abdomen is supine with the thorax slightly elevated (10-20 degrees). The optimal probe position is in the right paramedian epigastric area in the longitudinal direction. In all the 10 patients we were able to diagnose pneumoperitoneum preoperatively by ultrasound assessment. PMID- 10476263 TI - Do peripheral blood counts have any prognostic value following trauma? AB - BACKGROUND: Both lymphocytosis and lymphopenia have been observed following trauma and each has been claimed to predict mortality. An understanding of the early temporal changes in leucocyte counts may help to explain why such discrepancies have been found. The purpose of this study was to determine the early serial changes in peripheral leucocyte counts following injury. METHODS: A whole blood cell counter was used to measure serial total leucocyte, granulocyte, lymphocyte and monocyte counts from 20 patients in the first 3 h following blunt injury. Four to six peripheral blood samples were taken from each subject and grouped into 20 min intervals. RESULTS: Granulocytosis, lymphocytosis and monocytosis were evident within 40 min of injury. A biphasic granulocyte and monocyte response was observed in the first 3 h following trauma. Lymphocytes showed a linear resolution towards normal (regression coefficient -0.022; p < 0.01) with some individuals developing a lymphopenia. No correlation with injury severity was observed. CONCLUSION: Rapid mobilisation and subsequent redistribution of leucocytes occurs early following injury. Temporal changes will affect any predictive value of circulating leucocytes and studies must be precise with respect to blood sampling time. PMID- 10476264 TI - Good outcomes of open pelvic fractures. AB - While pelvic fractures are common injuries, those which communicate with an open wound are much less commonly encountered. These injuries have been associated with high morbidity and mortality in previous studies. We investigated our experience with open pelvic fractures retrospectively over a 79-month period. A total of 17 patients were identified, representing 2.7% of all pelvic fractures seen during this time period. There were no fatalities among the 15 patients who were not in cardiac arrest on arrival. Associated injuries were present in all patients. Functional outcome was good in eight patients and fair in seven. Although open pelvic fractures are severe injuries, survival in our study was 100% in those not arriving in cardiac arrest. PMID- 10476265 TI - Abnormal scaphoid fat pad: is it a reliable sign of fracture scaphoid. AB - In a retrospective study, initial radiographs of 40 proven cases of scaphoid fractures and 40 cases where fractures of scaphoid were eventually excluded were examined in a 'blinded' fashion. In none of these radiographs was a fracture line demonstrated by people who did not know the outcome. In 4 patients with proven fracture, normal scaphoid fat pads were demonstrated; a 10% 'false negative' result and in 9 patients without fracture, abnormal scaphoid fat pads were seen; a 23% 'false positive' result. The authors conclude that the scaphoid fat pad sign is a poor predictor of scaphoid fracture, even though a positive scaphoid fat pad sign indicates that an underlying fracture is likely. PMID- 10476266 TI - An improved cementing technique for revision hip surgery after peri-prosthetic fractures. AB - Replacement of the femoral component by a long-stemmed implant is one possible solution to the management of periprosthetic femoral shaft fractures. A problem often encountered, due to the extensive disruption of the femoral canal, is difficulty in achieving adequate cement pressurisation. We have overcome this by applying a split 60 ml syringe sleeve around the femoral shaft and holding it in position with clamps. This method has been used on five patients of mean age 87.7 (range 82-94) years with fractures that occurred at an average of 7.2 (range 4 17) years after the primary hip surgery. Two of the five patients achieved independent mobility and all of the patients were sufficiently independent to be discharged from the acute orthopaedic unit. We hope this cheap, effective method of supporting the bone fragments, maintaining alignment and achieving an acceptable level of pressurisation will lead to a decrease in the inevitable morbidity and mortality associated with a problem which as a result of an ageing population is likely to become increasingly common. PMID- 10476267 TI - Parameters of multiple organ dysfunction fail to predict secondary amputation following limb salvage in multiply traumatized patients. AB - OBJECTIVE: The purpose of this retrospective analysis was to evaluate whether systemic parameters that are used to characterize multiple organ dysfunction could also be used to predict the optimal time for amputation in patients failing limb salvage surgery following severe extremity injury. METHODS: The principal criterion for the study group was a lower limb amputation following a type IIIb or IIIc open tibial shaft fracture in multiply traumatized patients. This group was then divided into one group of primary amputation (group A) and one group of secondary amputation (group B). Beside these groups a third group of total traumatic lower limb amputation was recruited (group C). Data analysis included demographics, injury severity according to the ISS, evaluation of the limb injury by three different salvage scores (HFS, MESS and NISSSA) and organ function monitoring by the Denver MOD Score over a 14-day period posttrauma or up to 7 days after secondary amputation. RESULTS: Within the period 1987-1997 a total of 15 patients were recruited for group A (primary amputation), 10 patients for group B (secondary amputation) and nine patients for group C (traumatic amputation). The MOD score was only positive for pulmonary dysfunction, also reflected by the Horovitz quotient, in those patients that died later in either group. Mortality was higher in group A (three out of 15) compared with group B (one out of 10), which may be due to a higher ISS in group A (mean ISS 28.2 vs. 21.0 of group B). Although the MOD score of all recovered patients revealed no significant difference between group A and B, secondary amputation resulted in significantly longer demand of mechanical ventilation. According to our results secondary amputation may lead to transiently decreased pulmonary function but does not necessarily end in multiple organ dysfunction. The need for amputation in failed limb salvage was not indicated by systemic parameters. CONCLUSION: The right time for secondary amputation in order to prevent subsequent pulmonary dysfunction cannot be predicted by parameters otherwise indicating organ dysfunction. As the risk of secondary amputation for developing pulmonary dysfunction apparently cannot be estimated the decision for amputation or limb salvage should be made initially after trauma and should be the definite one. PMID- 10476268 TI - Early continuous noninvasive haemodynamic monitoring after severe blunt trauma. AB - BACKGROUND: Invasive haemodynamic parameters obtained by pulmonary artery (PA) catheterization from survivors' patterns were reported to provide criteria for therapeutic goals in high-risk elective surgery and accidental injuries. This approach is limited because PA catheterization requires critical care conditions; however, noninvasive methods can provide early information anywhere in the hospital. OBJECTIVES: To evaluate the feasibility of using noninvasive haemodynamic monitoring of patients with severe blunt trauma immediately after emergency department (ED) admission and to describe the early time course of haemodynamic events in survivors and nonsurvivors of blunt trauma. SETTING: A large, academic, level-I trauma centre. DESIGN: Prospective, descriptive haemodynamic study. PATIENTS AND METHODS: 38 severely injured patients, 22 (58%) survivors and 16 (42%) nonsurvivors, with ISS > 15 were monitored by: (a) an improved thoracic bioelectric impedance device that estimated cardiac output noninvasively and continuously, (b) simultaneous arterial oxygen saturation by pulse oximetry, (c) noninvasive blood pressure measurement and (d) transcutaneous oxygen and carbon dioxide sensors. The patients were monitored as soon as possible upon arrival at the ED and continued during the first 24 h or more after admission. When the patient reached the ICU, monitoring by PA catheterization was undertaken to validate the noninvasive methods and for continued diagnostic evaluations. RESULTS: Cardiac output estimations by thermodilution and bioimpedance were well correlated; r = 0.91. Survivors started with high cardiac index (CI) values that subsequently rose to over 4 L/min/m2; arterial oxygen saturation (SaO2), transcutaneous oxygen tension and transcutaneous-oxygen tension-to-inspired-fraction-of-oxygen-concentr ati on (PtcO2/FiO2) values were normal in survivors and higher than those of the nonsurvivors. In the 1st h after admission, nonsurvivors' blood pressures were higher than normal and higher than that of the survivors, but in the 2nd and 3rd h, both groups were in the normal range; thereafter, nonsurvivors' values were lower than survivors' and often lower than normal. CONCLUSIONS: The noninvasive haemodynamic monitoring system provides reasonably accurate, continuous, on-line, real-time display of haemodynamic data that show marked differences in the early patterns of survivors and nonsurvivors. The study suggests noninvasive monitoring may be used for early detection and correction of posttraumatic circulatory deficits. PMID- 10476269 TI - The assessment of acute knee injuries by Senior House Officers in the Accident and Emergency department. AB - This study in a large inner city Accident & Emergency (A&E) department investigated the awareness of new Senior House Officers (SHO's) of the major symptoms and signs of significant trauma to the knee. They were able to identify a knee effusion clinically but were unable to differentiate a haemarthrosis from a reactive effusion. An acute haemarthrosis is suggestive of significant damage such as fracture, cruciate ligament rupture, major meniscal tear, synovial tear or patella dislocation (Maffulli N, Binfield PM, King JB, Good CJ. J Bone Joint Surg [Br] 1993;75(6):945-949 [1]; Noyes FR, Basset RW, Grood ES, Butler DL. J Bone Joint Surg [Am] 1980;62-A:687-695 [2]; Jain AS, Swanson AJG, Murdoch G. Injury 1983;15:178-181.] [3]). In many cases a haemarthrosis may be the only abnormality demonstrable in the acute setting (Dehaven KE. Am J Sports Med 1980;8:9-14 [4]; Visuri T, Koskenvuo M, Dahlstrom S. Milit Med 1993;158(6):378 381 [5]). If this diagnosis is missed then patients may be inappropriately sent home without suitable treatment and follow up. Greater emphasis should be placed on the differentiation between reactive effusion and haemarthrosis in the knee during the teaching of orthopaedics both at medical school and also on arrival at the A&E department. PMID- 10476270 TI - Early failure of spiral blade using with an unreamed femoral nail: a case report. PMID- 10476271 TI - Bilateral upper ureteric disruptions caused by a traffic accident. PMID- 10476272 TI - Parallel twin blade technique for scar revision. PMID- 10476273 TI - AO/ASIF unreamed femoral nail: a minimally invasive approach to its insertion. PMID- 10476274 TI - Is routine removal of intramedullary nails justified. AB - This study investigates the outcome of 100 cases of lower limb intramedullary nail removal. 12 femoral and 25 tibial nails were removed for pain and 4 tibial nails for infection. For 22 tibial nails and 38 femoral nails there was no recorded indication. There where 3 operative complications, 2 abandoned procedures and 1 tibial fracture (nail extraction without prior removal of the distal locking screw). In 9 out of 16 cases anterior knee pain improved after tibial nail removal. 4 patients, previously asymptomatic, developed anterior knee pain following tibial nail removal. 40 patients who were discharged without crutches returned in pain and had to be given crutches to alleviate leg pain. 62 patients took a mean of 11 days sick leave. We conclude that all complications were avoidable. Intramedullary nail removal is safe. Patients with anterior knee pain should be told that their pain may persist and that knee pain may even arise. Most patients will require crutches and an average of 2 weeks away from work. PMID- 10476275 TI - The unreamed tibial nail in the treatment of distal metaphyseal fractures. AB - In recent years biological surgical solutions have been recommended in cases of distal tibial fractures, with the aim of reducing damage to soft tissues and to bone vascular supply. Between the years 1991-1995, 52 patients suffering from fractures of the distal tibial metaphysis were treated in our department with an unreamed tibial nail (UTN). Fractures were categorized in accordance with the AO Classification. 32 fractures had no articular involvement (43A1, 43A2, 43A3) and 20 included intra-articular extension (43C1, 43C2). 32 fractures had significant metaphyseal comminution (43A2, 43A3, 43C2). 12 were open fractures. All the fractures were treated by means of UTN using distal locking. In 13 patients an additional percutaneous interfragmentary fixation was also applied. 22 patients underwent an additional operation in order to facilitate bone union (dynamization, bone grafting and/or fibulectomy). In 50 of the 52 patients the fracture united with a very good range of knee and ankle motion. In 2 patients non-union with breakage of the UTN occurred and in two open fractures post operative infections were observed. Our experience with the treatment of fractures of the distal tibia, including intra-articular fractures with no comminution, points at an excellent functional outcome with a low incidence of complications. PMID- 10476276 TI - Implant failure of the gamma nail. AB - The gamma nail is a temporary implant characterised by a limited life expectancy under continuous dynamic stress. We reviewed a series of 839 patients with gamma nail stabilisation and found two fatigue fractures (0.2%) at the aperture of the distal locking holes. This complication has not been described in the literature. Metallurgic and scanning electron microscopic examinations proved that the fatigue zones occurred at the clover-leaf grooves, which is where the diameter of the gamma nail is reduced. The clover-leaf diameter is of no biomechanical use in gamma nail stabilisation. We suggested product modification of the gamma nail to produce implants with a round diameter instead of a clover-leaf shape. A modified implant is already in use at our institution. PMID- 10476277 TI - Hospital separation rates from osteoporotic and non-osteoporotic fractures in metropolitan and rural Australia. AB - Rural populations of the United Kingdom and Scandinavia have lower hospital discharge and incidence rates for hip fractures than urban ones. We compared hospital separations from fractures in rural health regions of Victoria, Australia, with those in metropolitan regions. Fractures were grouped into those commonly (1A), often (1B) or not often (2) associated with osteoporosis. For group 1A fractures, we found rural separation rates were significantly higher among both men and women who were aged either 20-24 or 45-64. The rural separation rates for group 1B fractures were also significantly higher in these age groups but only among men. For group 2 fractures, rural separation rates were significantly higher among young adults. Fractures of the neck, trunk, radius and ulna, but not neck of the femur, contributed to the rural excess of group 1A fractures. If rural adults are not hospitalised more often, risk factors for group 1 and group 2 fractures, which in the middle aged includes osteoporosis, may be more prevalent in rural Victoria. Our findings suggest that international studies of rural-urban differences in fracture rates are needed which include a range of anatomical sites. PMID- 10476278 TI - Conservative treatment of displaced fractures of the olecranon in the elderly. AB - We have analysed the functional results of the conservative treatment of displaced olecranon fractures in 12 patients with a mean age of 81.8 (73-90) years. The patients were evaluated at the time of the final follow up which was 15.2 (6-33) months. Physical examination and radiographic study were performed. None of the patients were limited in their daily activities. 67% of the patients were asymptomatic with an acceptable range of motion of their elbows. The clinical results were good in 8 cases, fair in 3 and poor in one, in spite of 9 cases of pseudoarthrosis. The grade of satisfaction was excellent in 11 patients and bad in only one. Conservative treatment of displaced olecranon fractures provides satisfactory results in old patients. PMID- 10476279 TI - Looking at trauma and deaths: Diyarbakir city in Turkey. AB - There were 329 trauma related deaths in Diyarbakir in 1997. Of these 226 were male (69%) and 103 were female (31%). The median age was 20 years old (range 1-82 years). Of the deaths, 30.5% were under 10, 51% were under 20 and 67% were under 30 years old. Two hundred and eleven deaths occurred in the hospital while 118 deaths occurred prehospital. Seventy-seven percent of hospital deaths (191) occurred in the first day. The most common cause of death was multiple injuries (151, 46%). Head injuries were the main reason for 128 deaths (46%). The most common mechanism of death was motor vehicle accident (131, 40%). The second was falls from a residential building (117, 33.7%). PMID- 10476280 TI - External fixation for open proximal ulnar fractures. AB - We describe our experience with the use of a pyramidal configuration external fixator in the management of open fractures of the proximal ulna. A prospective study was conducted. 21 patients with open fractures of the proximal ulna (Gustilo-Anderson Grade II and III A) were treated by the application of a new fixator frame. The soft tissue injury was managed with the help of the plastic surgeons. All the patients were followed up clinically and radiologically for a total duration of 3 years after the injury. They were rated on a combined objective and subjective assessment scale by an independent observer. All the patients were rated as good to excellent at the end of a 3 year follow up. The main advantages were simplified wound care and ease of supplementary plastic procedures. No vascular or neurological complications were encountered. The average time taken for union with the fixator was 9 weeks. All the fractures united well. Mobility and function of the elbow were preserved in all cases. PMID- 10476281 TI - Femoral shaft fractures in children treated by closed reduction and early spica cast with incorporated supracondylar Kirschner wires: a long-term follow-up results. AB - 38 children between 1 and 10 years of age with femoral shaft fractures were treated by closed reduction and early spica cast with incorporated supracondylar Kirschner wires and followed up prospectively. Overrides of maximum 15 mm, medial and anterior angulations less than 15 degrees and lateral angulation up to 5 degrees were considered acceptable. Posterior angulation and malrotation were not accepted. Of the 38 children examined at a mean of 65.6 months (range, 58-80 months) after fracture, none had any residual skeletal deformity and joint stiffness. At long-term follow-up, maximal shortening was 11 mm and overgrowth was 6 mm. The only factor associated with unacceptable shortening was shortening > 15 mm at the time of spica cast application. We believe that close follow-up during the first 3 weeks after cast application is important in order to achieve an acceptable final outcome. On the other hand this method of treatment is simple, safe and effective. It dramatically decreases hospital stay and cost of treatment and allows rapid return of the patients to their family environment. PMID- 10476283 TI - A novel method of incorporating windows in a cast. PMID- 10476282 TI - Primary surgical treatment of war injuries of major joints of the limbs. AB - We present the results of primary surgical treatment of war injuries of major joints in 339 patients, wounded in the territory of the former Yugoslavia in the period June 1991-October 1995. The total number of surgically treated joints was 358. The injuries were inflicted by the fragments of explosive devices in 176 (51.9%) patients and by bullets in 125 (36.9%) patients. A single wound was present in 160 (47.2%) and several wounds in 176 (51.9%) patients; 276 (77.1%) patients had injuries of articular bone. The most frequently injured joints were knee joint 167 (46.6%) and elbow joint 72 (20.1%). Associated neurovascular injuries were present in 68 (18.9%) patients. Postoperative bone fragment stabilization was required in 254 (70.9%) joints. Stabilization was by plaster of Paris in 148 (58.2%) and external fixation in 85 (33.4%) patients. Postoperative course was uneventful in 262 (77.3%) while postoperative complications occurred in 77 (22.7%) patients. Amputations were performed in 15 (4.4%) patients. After the primary surgical treatment 254 (74.9%) patients were discharged and directed to rehabilitation or home care and 81 (23.9%) patients required additional reconstructive surgery for bone and soft tissue defects. PMID- 10476284 TI - Use of knitted oxidized cellulose (Nu-knit) for the definitive packing of grade III liver fracture. PMID- 10476285 TI - Sesamoid displacement: a rare cause of 'clicking thumb'. PMID- 10476286 TI - Attitudes towards protective eye-wear: ocular trauma from lawn mowers. PMID- 10476287 TI - Scapular neck fractures; an update of the concept of floating shoulder. PMID- 10476288 TI - Unusual distal radial epiphyseal injury in a child. PMID- 10476289 TI - LeMaire procedure for anterior cruciate instability. PMID- 10476290 TI - A safe technique for distal biceps repair using a suture anchor and a limited anterior approach. AB - We describe a safe and simple method of repairing an acute rupture of the distal biceps tendon using a single limited (3 cm) anterior approach, a suture anchor, and the use of a plastic sheath (arthroscopy cannula or barrel of a 2 ml syringe) to protect the adjacent neural structures and to remove the need for extensive dissection and retraction in order to protect these structures. We have used this technique on three patients to date and all have regained a full range of flexion and extension at the elbow and pronation and supination of the forearm at a minimum of 6 months follow-up. There have been no neurovascular complications. PMID- 10476291 TI - Trauma in pregnant women: analysis of maternal and fetal mortality. AB - Twenty-seven traumatised pregnant women were analysed retrospectively over a period of 9 years. Mean age was 23.7 years (16-42 years). Gestational age ranged from 10 to 40 weeks (mean, 21.5 weeks), with most victims (46.1%) being in the second trimester. The predominant mechanism (65.3%) was blunt abdominal injury due to an automobile accident (the patient being run over or collision). At admission, 8 (30.7%) patients had haemodynamic alterations. 6 patients (23.0%) presented vaginal bleeding and 4 of these were haemodynamically normal. We analysed maternal mortality, fetal mortality and their causes. We also compared the median RTS and TRISS values for the groups with maternal-fetal survival and the group with maternal-fetal death. Fetal death occurred in all pregnant women admitted with vaginal bleeding. Maternal mortality due to haemorrhagic shock was 11.5%. Fetal mortality was 30.7%, with 37.5% of these deaths being caused by maternal death. The major cause of fetal mortality was a detached placenta (50.0%). The trauma indices, RTS and TRISS, were significantly lower (p = 0.0025 and p < 0.0001) in the group of maternal-fetal death but they were not of prognostic value in terms of fetal mortality. PMID- 10476292 TI - Exchange nailing for femoral shaft aseptic non-union. AB - Although rare, non-union of femoral shaft fractures is a cause of significant morbidity. In aseptic non-union, excellent union rates have historically been reported following reamed exchange femoral nailing. However, recently, a high incidence of failure requiring additional procedures has been reported. In light of these concerns and a recent change in our practice to the use of thin solid nails we undertook a retrospective study to determine the efficacy of exchange nailing with these modern nails in our hands. We reviewed records and radiographs of 25 patients who had a reamed exchange femoral nailing for established aseptic non-union. 24 patients (96%) united after exchange without the need for an additional procedure. The mean time to union was 29.75 weeks. Patients who had open bone grafting performed at the same procedure tended to unite quicker, but this did not achieve statistical significance (p = 0.14). Union times were not affected by smoking habits or nail type. This study demonstrates that reamed exchange nailing for aseptic femoral non-union remains an effective treatment. We believe that the nail type is less important than the biological effects of reaming, bone grafting and dynamization. PMID- 10476293 TI - The unstable distal radial fracture one year post Kapandji intrafocal pinning. AB - Kapandji pinning has been proposed as the treatment of choice for unstable Colles' fractures. The aim of this paper is to evaluate our experience treating unstable Colles' type fractures using this technique. Over a nine month period, 36 patients with Colles' type fractures were treated operatively at St. Vincent's Hospital. 22 of these fractures were deemed unstable and were treated using percutaneous intrafocal Kapandji pinning. 20 of these patients were recalled for review at a mean of 11.3 month post injury. At this stage the wrist was examined clinically and radiologically. Initial satisfactory correction of deformity was achieved by this technique. Between the time of wire removal and final review, however, there was significant recurrence of dorsal angulation (P < 0.05), but no significant radial shortening on radiographs. The patients had a satisfactory clinical result in spite of these radiological parameters. PMID- 10476294 TI - The Mennen plate: unsuitable for elderly femoral peri-prosthetic fractures. AB - The Mennen plate has been advocated for femoral shaft and femoral peri-prosthetic fractures. We utilised this fixation system in five patients with peri-prosthetic femoral shaft fractures. The operative technique was that described by the CMW laboratories. The postoperative regimen deviated from that recommended, i.e. prolonged bed-rest, since the patients were elderly and complicated by chest problems and pressure sores. Once the surgical wounds had healed, at an average of 11 days (range 10-14 days), one patient achieved non-weight-bearing mobilisation. Four patients were managed with bed-to-chair transfer only. All five plates failed at an average of 32 days (range 15-42 days). Only one patient had an identifiable accident to account for the failure. Two patients underwent revision hip arthroplasty and two had revision fixation with Dall-Miles (Howmedica), and Cable Ready (Zimmer) plate and cables. Femoral shaft peri prosthetic fractures are not adequately stabilised with the Mennen plate system, and prolonged recumbency in such an elderly population often worsens pre-existing medical problems. We advocate the use of one of the many available plate and cable fixation systems for such fractures in the elderly and in those in whom revision arthroplasty may not be possible. PMID- 10476295 TI - Peri-prosthetic femoral shaft fractures treated with plate and cable fixation. AB - Management of post-operative peri-prosthetic femoral shaft fractures remains difficult and controversial, with no widely accepted form of treatment. Fractures at the tip of a femoral component are associated with poor union rates. We present a series of 15 fractures at the component tip extending proximally in association with a loose cement mantle. All 15 patients were treated with open reduction and internal fixation using a plate and cable system. They were followed to clinical and radiological union, at an average of 3.5 months (range 3 5 months). At final review examination, 13 patients had achieved pre-fracture mobility, with one patient requiring a single walking stick, and one requiring a Zimmer frame. Three patients under-went revision surgery after successful fracture union and rehabilitation, with long stemmed femoral components. The plate and cable procedure is not technically demanding and avoids hip exposure. This form of internal fixation provides immediate fracture stability, allowing early ambulation, hence avoiding the complications and difficulties of other methods of treatment. Such fractures should be acutely treated to achieve fracture union with a plate, cable, and screw internal fixation technique. The loose components can be revised electively, preferably by a revision hip surgeon. PMID- 10476296 TI - Expectant management of haemoperitoneum. AB - Nonoperative management was successful in 81 of the 90 patients including two patients with penetrating injury. Nine patients were operated on after an initial period of observation, eight because of continued bleeding and one because of features of peritonitis. There were four deaths. These were due to multisystem injuries and not directly related to haemoperitoneum alone. Nonoperative management is a relatively safe and effective method in management of selected cases of haemoperitoneum. PMID- 10476297 TI - Admission level of Gc-globulin predicts outcome after multiple trauma. AB - BACKGROUND: Actin is the dominating protein in mammalian cells. Release of excessive amounts of actin into the circulation may result in a condition resembling multiple organ failure. The purpose of this study was to determine if admission levels of Gc-globulin can predict survival after multiple trauma. Also, we wanted to compare the predictive ability of Gc-globulin with that of the TRISS Like scoring system. METHODS: Fifty-seven patients with a median ISS 18 (16-75) were included. All patients had a blood sample taken median 42 min after the injury (19-110 min). Serum Gc-globulin was measured by rocket immunoelectrophoresis. RESULTS: On admission, all patients had significantly reduced levels of Gc-globulin compared with normal controls. Gc-globulin was significantly higher in the group of survivors (n = 41), compared with non survivors (n = 16). Median 237 mg/l vs. 188 mg/l (P < 0.01). The predictive ability of Gc-globulin regarding death was similar to that of TRISS-Like with positive predictive values of 69%, a negative predictive value of 84%, a sensitivity of 56% and a specificity of 90%. CONCLUSIONS: The predictive value of Gc-globulin regarding survival was similar to that of an established scoring system. Gc-globulin, alone or in combination with other parameters, may serve as a routine tool for early identification of patients at risk after severe injury, increasing the possibility of early intervention. PMID- 10476298 TI - Parachuting for charity: is it worth the money? A 5-year audit of parachute injuries in Tayside and the cost to the NHS. AB - All parachute injuries from two local parachute centres over a 5-year period were analysed. Of 174 patients with injuries of varying severity, 94% were first-time charity-parachutists. The injury rate in charity-parachutists was 11% at an average cost of 3751 Pounds per casualty. Sixty-three percent of casualties who were charity-parachutists required hospital admission, representing a serious injury rate of 7%, at an average cost of 5781 Pounds per patient. The amount raised per person for charity was 30 Pounds. Each pound raised for charity cost the NHS 13.75 Pounds in return. Parachuting for charity costs more money than it raises, carries a high risk of serious personal injury and places a significant burden on health resources. PMID- 10476299 TI - Tibial spine fractures: an analysis of outcome in surgically treated type III injuries. AB - We analysed the outcome of open reduction and internal fixation of type III tibial spine fractures, assessing treatment and determining a treatment protocol. A total of 10 patients presented over 3 years to our institution with a mean age of 15 years (range 10-21), a male-to-female ratio of 8:2. left to right 6:4 and anterior to posterior spine fracture 9:1. Only one patient had associated meniscal injury noted at arthroscopy (no treatment required). The mode of injury was road traffic accidents four, sports injuries three and falls three. The mean follow-up was 9 months. There were seven excellent results and three good results. Those patients with good results exhibited either minimal quadriceps weakness, extensor lag (< 10 degrees) or antero-posterior laxity. This reflects the experience of other authors in dealing with these injuries in younger patients. There is widespread agreement that types I and II should be treated by plaster cast alone and that is also the policy at our institution. We recommend a routine treatment protocol in type III injuries of (1) examination under anaesthesia, (2) arthroscopy (evaluating the fracture, cruciate integrity and other associated injuries), (3) open reduction and screw fixation and (4) vigorous physiotherapy/rehabilitation of all type III fractures, as we feel this provides the best possible outcome in these injuries. PMID- 10476300 TI - What are the requirements for medical cover at motor racing circuits? AB - Motor racing is perceived as a dangerous sport but few data are available on the incidence and nature of injuries sustained. The medical service requirement at one regional motor racing circuit was assessed by determining the incidence of injuries, the medical interventions required and the need for hospital referral and admission over a 5-year period. Five hundred and twenty-one patients, including support staff and spectators, attended the medical centre, of whom 14% were referred to hospital and 4% required admission. Each competitor had a 4% chance of requiring on-circuit medical attention, 0.6% chance of hospital referral and 0.17% chance of admission per race. Most major accidents involved more than two drivers. Twenty sustained major trauma including five pelvic fractures and two intraabdominal haemorrhages. Emergency intervention included intubation and ventilation in five. There were three deaths from a total of 9000 competitors (mortality rate 0.033%). This study shows that despite the nature of the sport, the mortality rate remains low with prompt skilled medical intervention. Medical personnel should include those competent in dealing with minor medical complaints as well as those with advanced airway management and resuscitation skills. Although national motor sport guidelines recommend a minimum of two attending doctors this would have been insufficient for multivehicle accidents. PMID- 10476301 TI - Simultaneous rupture of both peroneal tendons after corticosteroid injection: operative treatment. PMID- 10476302 TI - Skin burn associated with paint on synthetic plaster cast. PMID- 10476303 TI - Abdominal aortic rupture in a child after blunt trauma on a soccer field. PMID- 10476304 TI - Arterial injury complicating femoral shaft fracture: control of primary and secondary profunda femoral bleeding by transcatheter embolisation. PMID- 10476305 TI - Inflatable trays for the prevention of contamination in surgery. PMID- 10476306 TI - Raised intracompartmental pressures and compartment syndromes. PMID- 10476307 TI - Sleep disorders in older people. AB - Sleep complaints are common among older people. As there are often multiple contributing factors, insomnia should be considered a symptom, and not a diagnosis. There is a high prevalence of sleep apnea and nocturnal myoclonus. When these primary sleep disorders are suspected, the patient should be referred for polysomnography. Use of hypnotics should be discouraged for chronic insomnia. More research is needed to clarify the role of light therapy and melatonin in the treatment of sleep disorders in older people. PMID- 10476308 TI - AKEntAnnos. The Sardinia Study of Extreme Longevity. AB - This paper describes an epidemiological study performed in all centenarians living in Sardinia, a large island located in the Mediterranean sea, 120 Km from the Italian coast. Due to its long-standing isolation, low immigration rate, high endogamy and rather uniform lifestyle, Sardinia offers an ideal setting in which to study the genetic traits associated with extreme longevity and successful aging. A total of 233 potentially eligible centenarians were traced in the entire territory. Of these, 66 died prior to being interviewed, 11 were not found and unknown, and 15 refused to be interviewed. A multidimensional home interview was administered to 141 centenarians, and an equivalent number of 60-year-old controls matched for gender and area of residence. Furthermore, 41 living siblings of the centenarians, and 41 age- and sex-matched controls for these siblings were also studied. The prevalence of centenarians was 13.56 per 100,000, and the female/male ratio was approximately 2. Prevalence and female/male ratio were consistent across the four Sardinian municipalities and are, respectively, higher and lower than those reported in other population-based surveys. A number of methodological problems confronted in doing the field work, and plans for future analysis of this rich dataset are discussed. PMID- 10476309 TI - Factors related to the length of in-hospital stay of geriatric patients. AB - The aim of this study was to evaluate factors predicting length of stay in hospital of geriatric patients. Study participants were 402 patients (183 males and 219 females) consecutively admitted to four geriatric wards located in Chieti, Perugia, Pescara and Prato, Italy. Information on potential predictors of length of stay was collected; in particular, we assessed the presence and severity of specific chronic medical conditions, level of physical function, cognitive status, and depressive symptoms. Moreover, information on family and social support was obtained. In general, participants were old, often cognitively impaired and physically disabled. Average length of stay ranged from 9.4 +/- 3.3 days (Perugia) to 14.1 +/- 7.2 days (Chieti), and was statistically different across centers (p < 0.001). None of the specific medical diagnoses was associated with different length of stay. However, higher comorbidity score (p < 0.001), living alone (p < 0.01), lower MMSE score (p = 0.03), and poor functional status (p = 0.05) were all associated with longer length of stay. When these variables were included in a multivariate model predicting length of stay, differences between centers were no longer statistically significant. Findings of this study show that specific medical diagnoses are not adequate instruments to estimate length of stay in geriatric units. Other assessment systems based on extension of the social network, comorbidity, and the cognitive as well as the functional level need to be developed. PMID- 10476310 TI - Orthostatic hypotension in Alzheimer's disease: result or cause of brain dysfunction? AB - In Alzheimer's disease (AD), an association was found between autonomic dysfunction and frontal hypoperfusion in brain during orthostatic testing. To ascertain whether frontal hypoperfusion is dependent on longitudinal effects of hemodynamic disturbances, or contributes to them, we studied the relationship between the presence of orthostatic hypotension (OH) and resting cerebral blood flow (CBF) in late stages of AD. Twelve women with senile dementia of Alzheimer type (SDAT), and 15 non-demented women (mean age 82.6 years, SD 3.8 vs 81.8 years, SD 3.5) were examined with the orthostatic test. Four of 12 patients with SDAT, and 9 controls had OH (defined as systolic blood pressure fall > or = 20 mmHg). CBF was determined under resting conditions using 600 Mbq 99mTc HMPAO single photon emission computerized tomography (SPECT), and quantified in cortical areas in relation to cerebellum. In patients with SDAT and OH, CBF was lower in frontal and parieto-frontal cortical areas than in SDAT patients without OH. The former group was younger and had a shorter dementia duration. No significant differences in CBF were observed between controls with vs without OH. No differences in SDAT patients with or without OH were observed in the Berger dementia scale or Katz' ADL index. No difference in incidence of symptoms related to autonomic disturbances (diarrhea, obstipation, dysphagia, vertigo) was observed in either the SDAT or control group with regard to OH presence. We conclude that during the course of AD, OH can contribute to frontal brain changes and may exacerbate the disease. The further involvement of frontal dysfunction in aggravating blood pressure dysregulation in the elderly is discussed. PMID- 10476311 TI - Independence in instrumental activities of daily living and its effect on mortality. AB - Factors relating to six-year mortality in a representative sample of seventy-year old Jerusalem residents (N = 605) were investigated using logistic regression techniques. Around 16.3% of the study population died during the six-year post interview period. Bivariate analysis found elevated mortality related to being male, having more than one IADL dysfunction, more than two ADL dysfunctions, financial problems, no social support in times of emergencies, bad self-rated health status, cognitive impairment, confinement to bed during the fortnight prior to interview, and lack of regular exercise. Logistic regression controlling for gender, various clinical diagnoses, financial state, social support and smoking status showed IADL (ROR = 4.57, 95% CI 1.51, 13.90), cognitive impairment (ROR = 3.99, 95% CI 1.85, 8.59) and having been bed-sick a week or more during the preceding fortnight (ROR = 6.60, 95% CI 1.00, 43.86) to be independent predictors of mortality. All persons who had a cognitive problem and were dysfunctional in more than two IADL categories, and 93.8% of persons who had been bed-sick and had more than one IADL dysfunction died during the study period. Combined measures of these three easily obtainable variables could prove a cheap and efficient method of identifying at-risk elderly persons in order to provide them with specific programs aimed at decreasing functional decline, and hence mortality. PMID- 10476312 TI - Reduced glucose effectiveness as a feature of glucose intolerance: evidence in elderly type-2 diabetic subjects. AB - One of the factors determining glucose tolerance is glucose disappearance independent from the dynamic insulin (glucose effectiveness); the debate on its role in the development of Type-2 diabetes is still open. The aim of the present study was to evaluate insulin delivery, insulin sensitivity (SI), and glucose effectiveness (SG) in a group of elderly Type-2 diabetic patients (D, 4/6 F/M, age 67 +/- 2 years, 64 +/- 2 kg, BMI 23.8 +/- 0.5 kg/m2), compared to young controls (C, 4/6 F/M, 25 +/- 2 years, 72 +/- 4 kg, 23.7 +/- 1.1 kg/m2) and elderly controls (E, 2/4 F/M, 73 +/- 3 years, 63 +/- 4 kg, 23.1 +/- 0.5 kg/m2). We performed oral (OGTT) and intravenous (FSIGT) glucose tolerance tests. The OGTT showed that C and E were normotolerant, while D had a markedly reduced glucose tolerance. This was also confirmed in the FSIGT where the glucose tolerance index (KG) was 0.6 +/- 0.1% min-1 in D vs 1.8 +/- 0.2 in C and 1.5 +/- 0.2 in E (p < 0.0002). Total insulin area of D and the overall insulin delivery were not different from those of the control groups. The early phase area was instead significantly reduced (0.19 +/- 0.02 mU min/mL vs 0.61 +/- 0.06 of C and 0.46 +/- 0.06 of E, p < 0.001) given the reduction in the dynamic first-phase insulin delivery (0.86 +/- 0.17 min(microU/mL)/(mg/dL) vs 3.95 +/- 0.61 in C (p < 0.005) and 2.61 +/- 0.66 (p < 0.001) in E). SI of D was 3.4 +/- 0.4 10(-4) min 1/(microU/mL), not different from that of C (4.7 +/- 0.6) and E (3.5 +/- 0.2). This study showed a marked difference between SG of D and that of both control groups [0.010 +/- 0.001 min-1 vs 0.026 +/- 0.004 (p < 0.001) of C and 0.020 +/- 0.003 (p < 0.002) of E], mostly due to the zero-insulin component GEZI which was 0.006 +/- 0.001 in D vs 0.021 +/- 0.004 in C and 0.016 +/- 0.003 in E (p < 0.003). In the elderly groups, when taken together, SG exhibited a positive correlation with the area under insulin concentration during the early phase and with KG (r = 0.69, p = 0.0032 and r = 0.90, p = 0.0001, respectively), demonstrating the importance of the first-phase insulin delivery in modulating glucose effectiveness and glucose tolerance. PMID- 10476313 TI - Color discrimination, color naming and color preferences in 80-year olds. AB - The aim of the present study was to investigate color discrimination, color naming and color preference in a random sample of 80-year-old men and women. Knowledge of color perception in old age can be of value when using color contrast, cues and codes in the environment to promote orientation and function. The color naming test indicated that the colors white, black, yellow, red, blue and green promoted recognition to the highest degree among all subjects. A gender related difference, in favor of women, occurred in naming five of the mixed colors. Women also used more varied color names than men. Color discrimination was easier in the red and yellow area than in the blue and green area. This result correlates positively with visual function on far sight, and negatively with diagnosis of a cataract. The preference order for seven colors put blue, green and red at the top, and brown at the bottom, hence agreeing with earlier studies, and indicating that the preference order for colors remains relatively stable also in old age. This result should be considered when designing environments for old people. PMID- 10476314 TI - Drug use in homes for the aged. A comparison between mentally intact and mentally impaired residents. AB - The aim of this study was to analyze drug use in 347 residents in homes for the aged in Bergen, Norway. All drugs prescribed on a regular schedule were assessed, the prevalence of potentially harmful drug combinations, and inappropriately prescribed drugs were studied and related to mental capacity of the residents. Mental capacity was assessed by means of the Clinical Dementia Rating scale (CDR). The median number of drugs used was 4.0 (range 0-11, 95% CI 3.0-4.0). Mentally impaired residents consumed fewer drugs than mentally intact ones. In a logistic regression analysis the use of NSAIDS, beta-blockers and anxiolytics was significantly lower in mentally impaired residents (OR 0.37, 95% CI 0.17-0.80, OR 0.35, 95% CI 0.13-0.95, and OR 0.45, 95% CI 0.21-0.94, respectively), and the use of laxatives and vitamin B-12 higher (OR 2.19, 95% CI 1.04-4.62, and OR 5.08, 95% CI 1.11-23.25, respectively). Twenty percent of mentally intact and 21% of mentally impaired residents have potentially harmful two-by-two drug combinations, and drugs augmenting sedating properties were dominating. The prevalence of inappropriately used drugs was significantly lower in mentally impaired residents (20%) than in mentally intact ones (33%). Antidepressants with anticholinergic properties, benzodiazepines with long half-life and phenothiazines were the most frequent. PMID- 10476315 TI - Disability is associated with malnutrition in institutionalized elderly people. The I.R.A. Study. Istituto di Riposo per Anziani. AB - Several factors, such as disability, malnutrition, weight loss, and the interactive effect of diseases and aging have been associated with morbidity and mortality in the elderly population. Nevertheless, the relationship between disability and biological parameters has not been extensively investigated as a primary focus. In a cross sectional survey, 344 institutionalized elderly subjects were evaluated. Disability was measured according to the Katz index, and patients were divided into three groups: low (0-1 lost ADL), mild (2-4 lost ADL), and severe (5-6 lost ADL). Anthropometric, metabolic, and nutritional parameters were assessed; age, gender, number of pathologies, and number of drugs were also recorded. Data were analyzed by multiple comparison of means according to Scheffe, and by multivariate logistic regression analysis. An impairment in functional status was associated with several modifications in biological parameters. Logistic regression analysis showed that severe disability (5-6 lost ADL) was associated with low waist/hip ratio (< 0.9 vs > 0.9, OR: 1.56, CI 95%: 1.08-2.25), high body resistance (> 625 vs < 575 omega, OR: 1.39, CI 95%: 1.38 1.39), low plasma albumin levels (< 3.5 vs > 4.0 g/dL, OR: 6.02, CI 95%: 5.18 6.85), and low plasma transferrin levels (< 200 vs > 250 mg/dL, OR: 5.47, CI 95%: 4.56-4.58) independently of age, gender, comorbidity, and other confounding factors. Our results indicate that severe disability in ADL is strongly associated with anthropometric and biohumoral parameters suggesting the presence of malnutrition. A careful evaluation of the nutritional state appears to be of primary importance, and efforts to improve nutritional status are needed in approaching disabled elderly patients. PMID- 10476316 TI - Vitamin D status, parathyroid function and femoral bone density in an elderly Swedish population living at home. AB - The aim of this study was to determine vitamin D status and bone mineral density (BMD) in elderly, independent Scandinavians. A cross-sectional examination was conducted in a sample of 104 subjects (mean age 84.5 years), for possible correlations among anthropometric data, 25-hydroxyvitamin D, 1,25 dihydroxyvitamin D, intact parathyroid hormone (PTH) and femoral neck BMD. Daily dietary calcium and vitamin D intakes were below the recommended levels. Five percent of the subjects were taking calcium, and 30% vitamin D supplements. Previous fragility fracture was reported in 30% of the men, and 55% of the women. Higher mean values of serum 25-hydroxyvitamin D (p = 0.03) and femoral neck BMD (p = 0.03) were recorded in subjects spending > or = 3 hours outdoors weekly. Independently of time spent outdoors, subjects taking daily supplements of vitamin D (on average 5 micrograms) had higher 25-hydroxyvitamin D (p < 0.001) levels, without significant changes in femoral neck BMD values. Serum levels of intact PTH (reference range 8-51 ng/L) were elevated in 41%, of which 5% had mild primary hyperparathyroidism. Serum levels of 25-hydroxyvitamin D (reference range 10-65 ng/mL) and 1,25-dihydroxyvitamin D (reference range 15-55 pg/mL) were below the reference ranges in 4% and 5% of the subjects, respectively. When serum levels of 25-hydroxyvitamin D were lower than approximately 30 ng/mL, the serum intact PTH values began to increase from a level of 43 pg/mL. This threshold most probably reflected a more relevant value of vitamin D insufficiency, indicating that 45% of our subjects rather than 4% actually had hypovitaminosis. Multiple regression analysis demonstrated femoral neck BMD to be significantly and positively associated with higher body mass index, male gender, no history of fragility fracture and 25-hydroxyvitamin D (R2 = 0.39). It is concluded that in this sample of healthy elderly people who regularly spend time outdoors, vitamin D levels leading to secondary hyperparathyroidism seem to be a major cause of osteoporosis. Correcting chronic dietary calcium deficiency is likely to eliminate another factor contributing to poor bone health. PMID- 10476317 TI - Chemokines in inflammatory states. AB - Chemokines probably mediate inflammation in asthma by acting on endothelial cells, alveolar cells, neutrophils, eosinophils, basophils, mast cells, monocytes, and lymphocytes, which are inhibited by corticosteroids. In 1995, we found that MCP-1 provokes mast cell aggregation and [3H]5HT-release in cultured mast cells. In another study, MCP-1 and RANTES revealed to have a potent chemoattractive effect on basophilic cells originating from the rat skin. In this inflammatory model, RANTES also attracted eosinophils and macrophages along with basophilic cells. The effect of RANTES on inducing HDC mRNA was dose dependent. MCP-1 and RANTES provoked histamine release in intradermal mast cells and prostaglandin D2 generation. These effects clearly show that RANTES and MCP-1 are mediators of acute inflammatory responses. In chronic inflammatory reactions, MCP 1 is also present as we show in a study recently published by our group. In this paper, we found that MCP-1, strongly mediates the recruitment of mononuclear cells in the granuloma formed by KMnO4. In addition, MCP-1 mediated a parasitic infection caused by Trichinella spiralis in mice. Our data strongly demonstrate that chemokines, such as RANTES and MCP-1, mediate acute inflammatory response. PMID- 10476318 TI - Complications of allergic rhinitis. AB - With unfortunate high frequency, clinicians consider allergic rhinitis to be more of a nuisance than an illness. When in fact, allergic rhinitis is not only a very common disease process, affecting up to a cumulative frequency of 42% of the U.S. population by age 40, but can lead to significant short-term and long-term medical complications. Poorly controlled symptoms of allergic rhinitis may contribute to sleep loss, secondary daytime fatigue, learning impairment, decreased overall cognitive functioning, decreased long-term productivity and decreased quality of life. Additionally, poorly controlled allergic rhinitis may also contribute to the development of other related disease processes including acute and chronic sinusitis, recurrence of nasal polyps, otitis media/otitis media with effusion, hearing impairment, abnormal craniofacial development, sleep apnea and related complications, aggravation of underlying asthma, and increased propensity to develop asthma. Treatment of allergic rhinitis with sedating antihistamine therapy may result in negative neuropsychiatric effects that contribute to some of these complications. Sedating antihistamines may also be dangerous to use in certain other settings such as driving or operating potentially dangerous machinery. In contrast nonsedating antihistamines have been demonstrated to result in improved performance in allergic rhinitis. PMID- 10476319 TI - Pharmacotherapy to prevent the complications of allergic rhinitis. AB - Allergic rhinitis is an immunologic disease with effects that extend beyond the symptoms that occur subsequent to allergen exposure. A reduced quality of life and medical conditions such as asthma, sinusitis and otitis media are well recognized complications of allergic rhinitis. Craniofacial abnormalities, nasal ployps, and respiratory infections have been linked to allergic rhinitis, but the evidence is conflicting. This article reviews the complications of allergic rhinitis, their prevalences, possible mechanisms for their relationship to allergic rhinitis, and the prevention of these complications via pharmacologic treatment of allergic rhinitis. PMID- 10476320 TI - The future of allergy practice: a different perspective from another academician. PMID- 10476321 TI - Unusual responses to contact allergens. PMID- 10476322 TI - Changing patterns in academic allergy-immunology. AB - In evaluation of the current Allergy-Immunology (AI) Program of the Department of Medicine at Northwestern University Medical School and in planning for the future, it appeared that our assessment of changes in the AI program since its inception might be of value to other AI academic programs. Further, we might receive suggestions from other academic AI programs, and we request such advice. PMID- 10476323 TI - Chronic allergic-like dermatopathies in nickel-sensitive patients. Results of dietary restrictions and challenge with nickel salts. AB - Nickel frequently contaminates foods. In sensitized patients, dietary nickel can cause a relapse of contact eczema and also widespread chronic dermatopathies quite similar to those triggered by authentic food allergy (IgE-mediated), from atopic dermatitis to chronic urticaria with angioedema. The present study was intended to evaluate the the results of an elimination diet and of the oral challenge test with nickel salts in a population of adults suffering from chronic urticaria or angioedema, pruritus or atopic dermatitis, and concomitant contract sensitization to nickel salts. The study involved a population of adult patients (112 subjects, 106 women and 6 men, aged from 16 to 58, mean age 29 +/- 10) with widespread allergic-like dermatopathies and contact sensitization to nickel salts (positive patch test). All of these subjects were prescribed a low nickel diet for four weeks. The patients who recovered or whose clinical manifestations greatly improved underwent an oral double-blind, placebo-controlled challenge: they were administered two successive, noncumulative doses of 10 and 20 mg nickel sulphate hexahydrate, respectively equal to 2.23 and 4.47 mg of elemental nickel. A search for specific IgE and the check on skin reactivity by skin-prick test against nickel were carried out in the patients who had shown particularly severe reactions after the oral challenge. A low nickel diet was effective in controlling the symptoms in 44 patients (39.28%, among whom there was one man). The oral double-blind, placebo-controlled challenge test was positive in all the patients who had favourably responded to the elimination diet, except one. In the patients with anaphylactoid reactions on the oral challenge, skin-prick tests were negative and no serum-specific IgE antibodies against nickel were found. Such findings appear to demonstrate that, in some patients with concomitant contact allergy, intolerance to ingested nickel salts might be the real cause of the onset and perpetuation of widespread, chronic, allergic-type dermatopathies. PMID- 10476324 TI - Upper airway epithelial cells support eosinophil survival in vitro through production of GM-CSF and prostaglandin E2: regulation by glucocorticoids and TNF alpha. AB - Production of GM-CSF by epithelial cells has been implicated in eosinophil survival within the airways, although GM-CSF promotes neutrophil and monocyte survival as well. Using primary cultures of human airway epithelial cells, we undertook a comprehensive examination of factors that enhance eosinophil survival or apoptosis. Unstimulated epithelial cells were compared to epithelial cells stimulated with TNF-alpha in the presence or absence of dexamethasone. A striking increase in survival was observed when peripheral blood eosinophils were cultured with supernatants derived from unstimulated and TNF-alpha-stimulated epithelial cells. Cultured epithelial cells were examined for transcripts of cytokines shown to enhance eosinophil survival (GM-CSF, IL-3, IL-5, IL-13, and IFN-gamma), and transcripts for cytokines promoting apoptosis (IL-10 and TGF-beta). GM-CSF transcripts, but not the other cytokines, were present in unstimulated epithelial cells, and levels were increased with TNF-alpha stimulation. TNF-alpha stimulation increased the levels of GM-CSF and PGE2 in epithelial cell supernatants and dexamethasone suppressed the TNF-alpha induced increases. The survival effects of the TNF-alpha-stimulated supernatants were effectively blocked by neutralizing antibodies to GM-CSF or by dexamethasone treatment of epithelial cells. Selectivity of GM-CSF for eosinophil versus neutrophil survival was demonstrated and suggests that epithelial cell regulation of GM-CSF and PGE2 contribute to eosinophil survival in vitro and may contribute to eosinophil accumulation in allergic disease. PMID- 10476325 TI - The use of inhaled corticosteroids in asthma. AB - Inhaled corticosteroids are the most important therapeutic agents for the pharmacological control of pulmonary inflammation in asthma. There is concern, however, about the occurrence of side effects with the long-term use of inhaled corticosteroids. Because of the potential seriousness of some of these side effects, patients should be monitored carefully and preventively treated for the side effects. Various noncorticosteroid medications have been recommended in guidelines as substitutes for inhaled corticosteroids for daily use as long-term controllers in asthma, e.g., sustained-release theophylline, long-acting beta agonists, leukotriene modifiers, cromolyn, and nedocromil. However, of the long term controller medications recommended in the guidelines, only inhaled corticosteroids have to date, been shown clinically to reduce asthma fatalities and to prevent asthma induced lung remodeling. PMID- 10476326 TI - Asthma among the famous. Moses Gunn (1929-1993) American actor. PMID- 10476327 TI - Asthma among the famous. Teresa Teng (1953-1995) Taiwanese vocalist. PMID- 10476328 TI - Health and the right to privacy. PMID- 10476329 TI - If I am only for myself, what am I? A communitarian look at the privacy stalemate. PMID- 10476330 TI - Direct contracts, data sharing and employee risk selection: new stakes for patient privacy in tomorrow's health insurance markets. PMID- 10476331 TI - Online without a net: physician-patient communication by electronic mail. PMID- 10476332 TI - Telemedicine and integrated health care delivery: compounding malpractice liability. PMID- 10476333 TI - Cyber-malpractice: legal exposure for cybermedicine. PMID- 10476334 TI - Informed consent in the electronic age. PMID- 10476335 TI - Reflective choice in health care: using information technology to present allocation options. PMID- 10476336 TI - Broadcasting clinical guidelines on the Internet: will physicians tune in? PMID- 10476337 TI - The siren song of the elderly: Florida's nursing homes and the dark side of Chapter 400. PMID- 10476338 TI - [Current status and trends in the development of medical information systems for morphological research]. AB - The time of cooperative action of the information medical systems is coming. Standardization of clinical laboratory information systems is one of the most difficult but important problems for clinical laboratory community and technicians. Electronic data exchange requires agreement on the data element format by which healthcare institutions can exchange. Computerization of health care service raises the problem of security because the risk of violation of medical privacy is dramatically increasing. Unauthorized users can access, copy, alter, delete or distort hundreds or thousands of medical records within minutes. Information can be faulted by individuals or system failure. It is necessary to discuss and make a final decision of joining some standard for the purpose of integration. PMID- 10476340 TI - [Hyperplastic changes and aldosterone content in the adrenal cortex in essential hypertension and primary aldosteronism]. AB - Autopsy and operative material (adrenalectomy for hyperaldosteronism) was studied to elucidate morphology, incidence of nodules, aldosterone content in the adrenal of patients with essential hypertension (EH). It was established than nodular masses in the adrenals in the form of micro and macronodules are present in 80% of EH patients. Aldosterone content in the adrenals in both nodules and in the adjacent cortex is significantly higher than in the adrenals of patients without EH. This fact as well as increased cell nuclei size in the fascicular and glomerular zones indicate high secretory activity of the adrenals in EH. Clinicomorphological comparisons in patients after adrenalectomy because of hyperaldosteronism syndrome allow to conclude that grave forms of EH may be followed by aldosteronism syndromes with nodular hyperplasia of the adrenal cortex being the basis of the syndrome. Indications to adrenalectomy require clear criteria of differential diagnosis with primary aldosteronism. PMID- 10476339 TI - [Changes in suprachiasmatic and paraventricular hypothalamic nuclei in essential hypertension]. AB - Suprachiasmatic and paraventricular hypothalamic nuclei (SCN and PVN, respectively) were studied in humans with essential hypertension (EH) and in healthy individuals who had normal blood pressure and died by accident (control group). Immunohistochemistry, hybridization in situ using computer image analysis have shown that EH patients have decreased number of vasopressin (VP) positive cells in SCN, high number of corticotropin-releasing hormone (CRH) producing neurones in PVN and increased amount of mRNA for CRH in them. A negative linear correlation was found between the number of CRH-producing cells in PVN, amount of mRNA for CRH in them and the number of VP-synthesizing cells in SCN. The presence of GABA in VP-producing cells in SCN together with the data obtained suggest the presence of certain "disinhibition" of CRH-producing cells in PVN in EH which could cause enhanced synthesis of ACTH in anterior hypophysis and increased secretion of corticosteroids by the adrenal gland. PMID- 10476341 TI - [Features of atherosclerosis in the ascending human aorta and the link between its development and the main risk factors of ischemic heart disease in patients after aortocoronary shunting]. AB - Atherosclerosis was found in all ascending aorta biopsies of 125 patients aged 42 to 65 years who underwent aortocoronary bypass surgery. Lipid spots only were found in 91.2% of cases, in 11 patients (8.8%) there were lipid plaques. Three main types of lipid spots were recognized: type I--primary extracellular lipoidosis (40.0%); type II--mainly intracellular lipoidosis (24.8%) and type III -cell lipoidosis with a pronounced component of secondary extracellular lipoidosis (26.4%). A specific feature of ascending aorta atherosclerosis is a frequent combination of intima lipoidosis with media lipoidosis: 92% in type I, 100% in type II and 93.9% in type III. This is probably an important way of lipid elimination from the intima and this predetermines a "mild" course of atherosclerosis in this part of the aorta. Types II and III of lipoidosis occurred more frequently in lipid metabolism disturbances and only under these conditions clusters of foam cells were observed. Arterial hypertension and smoking have a leading role in development of intimal hyperplasia. When assessing atherosclerosis activity in the operated patients not only risk factors of ischemic heart disease but also the results of aorta biopsies studies should be taken into consideration. PMID- 10476343 TI - [Status of the adrenal medulla in various forms of cardiomyopathy]. AB - Adrenal medulla was studied quantitatively by a histofluorescent method and by that of Wood in 20 persons who had died at the age 35-50 years. The autopsy material was obtained from 11 dead bodies (control group--accident death) and those who had dilated and alcoholic cardiomyopathy (DCMP and ACMP, respectively). Catecholamine luminescence intensity (CLI) was higher in ACMP compared to control. On the contrary, CLI was low in chromaffin adrenal tissue in DCMP. The histofluorescent results corresponded to those obtained by a light microscopy by Wood method. The differences in results concerning catecholamines obtained in ACMP and DCMP were probably due to the differences in their activities in patients when they were alive (low in DCMP and high in ACMP). PMID- 10476342 TI - [Clinico-morphologic results of laser myocardial revascularization in ischemic heart disease]. AB - Transmyocardial laser revascularization was applied in 58 patients with ischemic heart disease. The objective criterion of the method efficiency was an increase in myocardium perfusion due to newly formed blood vessels under influence of laser irradiation. The laser treatment has improved cardiac function and general condition of the patients. The surgery did not exert considerable influence on the contractile capacity of the myocardium in the patients treated. PMID- 10476344 TI - [Metabolism of androgens in various histological variants of bone tumors]. AB - The role of androgens in human normal and neoplastic bone tissues is still unclear. The paper presents data on metabolism of androgens in homogenates of malignant (osteosarcoma, chondrosarcoma, Ewing and giant cell) and benign primary tumors from 46 male and female patients aged 14-58 years. Using two substrates (testosterone and 5 alpha-dihydrotestosterone) for the first time are shown activities of main enzymes of androgen metabolism in all tumor types. 5 alpha reductase activity was similar in all tumors, 3 alpha-hydroxysteroid dehydrogenase activity was the highest while that of 17 beta-hydroxysteroid dehydrogenase was the lowest. Unknown metabolite(s) of 5 alpha dehydrotestosterone was discovered which may be hydroxy-metabolites of 5 alpha androstane-3 beta, 17 beta-diol. A principal scheme of androgen metabolism in human neoplastic tissue is proposed. PMID- 10476345 TI - [Adrenocortical tumors: mitotic index and nuclear size as criteria for differential diagnosis and prognosis]. AB - A comparative study of the mitotic index (MI) and karyometric indices in 15 adrenocortical tumors with clinical syndrome of hypercorticism allowed to distinguish between adenomas and carcinomas. Carcinomas with hypercorticoidism symptoms have common features with inactive carcinomas but are different by lower expression of malignancy criteria (polymorphism, atypia, necrotic foci, capsule and vessel invasion), higher MI, increasing deficiency of cell division and larger nuclear size of tumor cells. Such tumors may be included into the prognostic group of risk. PMID- 10476346 TI - [Stromal vessels of invasive ductal mammary gland carcinoma]. AB - With the use of antibodies against factor VIII, vascular bed (number, surface and caliber of vessels) was studied in 64 cases of invasive ductal carcinoma and in normal breast tissue. The trend to a decrease of the vessels number with growing size of the tumor and emergence of lymph node metastases was observed. No difference was found in the development of vascular bed depending on the degree of malignancy, clinical course, tumor or normal breast tissue. PMID- 10476347 TI - [27 Clinico-anatomic conferences of the Department of Pathology of the Clinics and Chair of Pathology, I.M. Sechenov Moscow Medical Academy for 1994-1997 (Protocols Nos. 383-409)]. AB - The majority of the conferences were topical with discussion of most urgent problems of clinical pathology. Analysis of each autopsy was preceded by the report of a leading expert in the subject discussed. PMID- 10476348 TI - [Modern computer data base "necropsy protocol": potential and prospects]. AB - Basic principles in developing computer variant of necropsy protocol with wide illustrative possibilities are presented. Perspectives in the use of this information base for research and practice and for teaching the staff of the research and clinical institutions are demonstrated. PMID- 10476349 TI - [Histologic tissue processing in an automated microwave histoprocessor]. AB - A new vacuum histoprocessor has been tested as well as new more effective protocols to work with this processor. The device allows histoprocessing of 50 samples simultaneously of surgery, biopsy or autopsy material during 1-1.5 h. Apart from the standard protocols, new protocols with a much lower (5-10 times) expenditure of the chemicals can be implemented due to combination of microwaves and vacuum. Each protocol was tested repeatedly (20 and more times) with stable good quality of microscopic image. PMID- 10476350 TI - [Pathomorphologic changes in the sympathetic portion of the autonomic nervous system and cardiovascular pathology]. AB - Neurohistochemical and ultrastructural study of the sympathetic innervation of the cardiovascular system indicated its important role in the age alterations and genesis of certain diseases. Early beginning (from the age of 30) of the involution of the heart adrenergic plexus is confirmed in healthy persons. Focal desympathization of the myocardium is detected in sudden cardiac death. A decrease of sympathetic plexus density in zones undergoing atherosclerotic changes is of importance in the genesis of atherosclerosis. In arterial hypertension a primary stage of sympathetic neurones hypertrophy is changed into the phase of the mediators exhaustion in the sympathetic plexus. Myocardiopathies are characterized by progressing myocardial desympathization. An important problem of cardiosurgery is methods of heart reinnervation at its transplantation. PMID- 10476351 TI - [Embryologic and pathogenetic aspects of the common atrioventricular canal development]. AB - Atrioventricular canal (AVC) is an inherited defect the embryological basis of which is deficiency of the affluent part of the interventricular septum (IVS). Folds of the atrioventricular (AV) valves are formed from the myocardium and not from the endocardial thickening but much later than the IVS formation. Under the conditions of the affluent part of IVS the mode of connection of the anterior fold of the left AV valve creates the narrowing of the left ventricular effluent part. Endocardial thickenings play a role of a glue fixing corresponding structural components of AV valves and primary heart partitions. The degree of sticking together determines great variants of the defect anatomy. Important deficiency of the affluent part of IVS is possible this making the function difficult due to space change of the endocardial thickenings. The common valve ring with freely floating bridge-like folds is frequently revealed in such case. The notion "deficiency of the endocardial thickenings" has the only manifestation as an isolated splitting of the anterior fold of the mitral valve and exhibits main features of AVC. PMID- 10476352 TI - [Helicobacter heilmannii in the pathology of the stomach and duodenum]. AB - Historical aspects of the discovery of the spiral-like non-spirochetal bacteria belonging to Helicobacter and called helicobacter heilmannii (Hh)--previous name Gastrospirillum hominis--and producing under some conditions, like Helicobacter pylori, analogous gastroduodenal pathology (chronic gastritis, gastric ulcer, MALT-lymphoma and stomach cancer) are presented. Microbiological and structural features of this agent are considered as well as its relations with an apical surface of epitheliocytes, difficulties in diagnosis and methods of optimization of the bacteria detection in the biopsy material. The data on a wide spread of Hh in many representatives of the animals and a proven information about transmission of this infection from domestic animals (dogs and cats) to humans are available. The causes of a weak resistance of these bacteria to standard eradication schemes are analysed. PMID- 10476353 TI - Oral candidiasis in children with immune suppression: clinical appearance and therapeutic considerations. AB - Children and adolescents with immune compromise and suppression are particularly susceptible to the development of oral candidiasis. In fact, oral candidiasis is the most common oral manifestation in HIV-infected children. Oral candidiasis has been linked to a depressed immune system, more rapid progression to AIDS, more advanced stage of disease in AIDS, and decreased survival. Several different forms of candidiasis may be recognized clinically. These forms are 1) pseudomembraneous candidiasis; 2) erythematous (atrophic) candidiasis; 3) papillary hyperplasia; 4) chronic hyperplastic candidiasis; 5) angular cheilitis; and 6) median rhomboid glossitis. Diagnosis of candidiasis is primarily based upon clinical appearance; in some cases, however, exfoliative cytology and/or biopsy of the lesion may be necessary. It is also possible to culture the lesion to determine the specific subtype of candidia and to evaluate the susceptibility of the fungus to specific antifungal agents. Both topical and systemic treatment by antifungal medications are readily available. PMID- 10476354 TI - Periodontitis in the child and adolescent. AB - Early onset periodontitis (EOP) which affects individuals thirty-five and younger is characterized by a rapid rate of bone loss and disease progression with defects in host response and a specific etiological microbial flora. Within this classification, there are three subsets of the disease: prepubertal periodontitis, juvenile periodontitis, and rapidly progressive periodontitis. The characteristics of each disease are described along with illustrative cases that will help clinicians diagnose EOP in their patients. Currently prescribed treatment modalities and guidelines from the literature are also discussed in this paper. EOP may represent a complex set of diseases that are likely to be reclassified as further advances in microbiological and genetic research are made. Clinicians should be aware of such changes in order to diagnose, treat, and refer their patients for comprehensive care. PMID- 10476355 TI - Cephalometric changes during self-correction of primary anterior crossbite. AB - Longitudinal growth changes were investigated cephalometrically for sixty-one Japanese girls who had anterior crossbite at the initial visit (ages 3 years and 11 months on an average). Cephalograms were taken annually from the initial visit to the end of the observation period (ages 7 years 6 months on an average). Eleven of them (Group N1) showed self-correction of anterior crossbite during the primary dentition; ten (Group N2) showed self-correction when the permanent incisors erupted and forty (Group R) did not show self-correction. There was no statistical skeletal difference between Groups N1 and N2 during the observation period. Groups N1 and N2 were larger, however, than Group R in A'-Ptm'/Pog'-Go (p < 0.05) and Group N1 was smaller than Group R in Pog'-Go/N-S (p < 0.01) at the end of the observation period. In summary, favorable growth of the maxilla relative to the mandible leads to self-correction of primary crossbite and when mandible size relative to the cranial base is large, primary crossbite does not show self-correction. PMID- 10476356 TI - The Innsbruck Sensorimotor Activator and Regulator (ISMAR): construction of an intraoral appliance to facilitate ingestive functions. AB - Oral sensorimotor therapy is practiced widely with children who have neuromotor impairments, such as cerebral palsy and eating problems. Although improvement in ingestive skills can be achieved in the short term (5 months), long-term effects (over 12 months) remain to be examined. Interventions with intraoral appliances are used in children with moderate impairments of the oral-motor system and offer an opportunity for long-term treatment. Instead of the daily oral sensorimotor exercises, which must be provided by a qualified therapist, the intraoral appliance is worn during the night, so that the "therapy" is initiated and controlled by the child. The purpose of this paper is to describe the appliance: its prescription, fabrication and therapeutic use. A case study illustrates that improvement in ingestive skills, efficiency of eating, and marked weight gain can be achieved. PMID- 10476357 TI - The effect of dental training on the reactions of mentally handicapped children's behavior in the dental office. AB - This study was planned to observe the negative reactions shown by mentally handicapped children during the dental examination and prophylaxis, and to confirm whether there was a difference in these reactions after dental training. Twenty mentally handicapped children (between 6-18 years) were examined by a dentist and their reactions were reported, before dental training was given. Later, a special educator and a dentist trained the children about the dental process. After the training the children were examined and their reactions were reevaluated. The results obtained from the first and the second appointments were compared using McNemar Test. There were significant differences between the first and the second results. PMID- 10476358 TI - Children with disabilities: more than just numbers. AB - Listing of the numbers of children with disabilities provide little to no insight in the difficulties faced by children with disabilities and their families. A review is provided of the emotional and psychological hurdles that need to be considered as pediatric dentists continue to provide the necessary services. PMID- 10476359 TI - A "dirty secret": the abuse of children with disabilities. AB - The media and professional and lay literature are replete with the tragic stories of the abuse and neglect of children. Limited references, however, are made to the even more unbelievable reality that children with disabilities are maltreated at a rate that may be as great as four to ten times that of children without disabilities. The characteristics that make children with disabilities more vulnerable to abuse, the nature of the abuse, the variation by gender, the financial costs of abuse and the reality that this abuse may occur in the "best of families" are reviewed in an effort to increase practitioner awareness of the "dirty secret." PMID- 10476360 TI - Regional odontodysplasia: report of a case involving two separate affected areas. AB - The purpose of this study was to report in a 12-year old white child a rare clinical condition called regional odontodysplasia. The diagnosis was based on clinical and radiographic findings. Due to the patient's concern over appearance, the treatment involved extracting the affected teeth and subsequently placing partial removable prostheses, thus providing the patient better function and appearance. The teeth that were removed were examined histologically, both in routine preparation stained by hematoxyline and eosin, and in ground sections. PMID- 10476361 TI - A study of the clinical, histopathologic and ultrastructural aspects of enamel agenesis: report of case. PMID- 10476362 TI - Incontinentia pigmenti (Bloch-Sulzberger syndrome): report of case. AB - Incontinentia Pigmenti is an inherited disorder with predominantly ectodermal abnormalities. The dental effects, delayed eruption, hypodontia, and microdontia, are very similar to anhidrotic ectodermal dysplasia. It is important that children with incontinentia pigmenti gain access to specialist dental care including pediatric dentistry, orthodontics, prosthodontics and oral surgery. PMID- 10476363 TI - Menkes disease and Wilson disease: two sides of the same copper coin. Part I: Menkes disease. PMID- 10476364 TI - Adaptive motor strategy for squatting in spastic diplegia. AB - Motor strategies, defined by kinetic, kinematic and/or muscle activation patterns, reflect neural planning of movement, which takes into account central as well as peripheral constraints. Major alteration is expected in cerebral palsy, a condition characterized by abnormal posture and movement secondary to early lesion of the brain. The objective of this study was to characterize the motor strategies involved in disruption of posture in cerebral palsy of the spastic diplegia type and compare them with normal controls. The optoelectronic ELITE system was used to record and analyse the movement of squatting from the standing position with the arms extended forward in 11 children with spastic diplegia aged between 3 and 12 years and 11 age-matched normal controls. Normal children maintained gaze and arm horizontality and trunk verticality throughout the movement. The knee followed an oblique trajectory. Its angular velocity profile showed a short, single-peaked, ascending phase. The onset of movement was preceded by deactivation of the semimembranous muscle. In diplegic children, gaze and arm horizontality and trunk verticality were lost. The ankle was rigidified, resulting in spatial fixation of the knee. The ascending phase of the knee velocity profile was prolonged and multi-peaked. There was widespread muscle co contraction from the outset of movement. No anticipatory deactivation was evidenced, but anticipatory bursts appeared in the soleus. Patients with cerebral palsy have to organize a limited motor repertoire from a restricted neural potential. Consequent motor strategies presently demonstrated in spastic diplegia are distinct and appear as an original alternative to those of normal subjects. PMID- 10476365 TI - Menkes disease: study of the mitochondrial respiratory chain in three cases. AB - Mitochondrial oxidative metabolism in three patients with typical Menkes disease was studied. In two cases, a general decrease in all of the respiratory chain complex activities (I, II, III and IV) was observed. However, in the most severe case, these activities were entirely normal. Our results emphasize the diversity of the cellular expression of Menkes disease which can, in some cases, be associated with a mitochondrial encephalomyopathy. PMID- 10476366 TI - Rapid onset dystonia-parkinsonism in a 14-year-old girl. AB - A painful dystonia of rapid onset and associated parkinsonian features is described in a girl aged 14 years. The condition is refractory to treatment and has led to severe neurological disability. Her father had presented with a similar picture. PMID- 10476367 TI - Botulinum toxin for amelioration of knee contracture in Duchenne muscular dystrophy. AB - An 11-year-old non-ambulant boy with Duchenne muscular dystrophy developed tightness in his left knee flexors, which caused difficulties in standing exercises. Botulinum toxin A (BTX-A) was injected into the medial and lateral hamstring muscles and the range of motion increased by 20 degrees but after 5 months, when the pharmacological effect of BTX-A had vanished, an increase of only 5 degrees in range compared with the initial finding was left. It is concluded that there may be a role for BTX-A in controlling contractures in Duchenne muscular dystrophy. PMID- 10476368 TI - Secondary supratentorial anaplastic astrocytoma following treatment of medulloblastoma. AB - The development of secondary tumours is a rare but well known late effect of radiation therapy of lesions in the central nervous system. Most radiation induced tumours are of mesenchymal origin, but on rare occasions gliomas can occur. We describe a patient in whom a supratentorial anaplastic astrocytoma developed 15 years after surgery and radiotherapy for a childhood posterior fossa medulloblastoma. A concise review of the pertinent literature is given. PMID- 10476369 TI - 1,3-Dichloropropene. PMID- 10476370 TI - 1,2-Dimethylhydrazine. PMID- 10476371 TI - Hydrazine. PMID- 10476372 TI - Isoprene. PMID- 10476373 TI - Isopropanol. PMID- 10476374 TI - Malonaldehyde (malondialdehyde). PMID- 10476375 TI - 4,4'-Methylenediphenyl diisocyanate and polymeric 4,4'-methylenediphenyl diisocyanate. PMID- 10476376 TI - Methyl methanesulfonate. PMID- 10476377 TI - 2-Nitropropane. PMID- 10476378 TI - 1,3-Propane sultone. PMID- 10476379 TI - beta-Propiolactone. PMID- 10476380 TI - Resorcinol. PMID- 10476381 TI - 1,1,1,2-Tetrachloroethane. PMID- 10476382 TI - Tetrafluoroethylene. PMID- 10476383 TI - 1,1,2-Trichloroethane. PMID- 10476384 TI - Vinylidene chloride. PMID- 10476385 TI - N-vinyl-2-pyrrolidone and polyvinyl pyrrolidone. PMID- 10476386 TI - Xylenes. PMID- 10476387 TI - Acetamide. PMID- 10476388 TI - Acrylic acid. PMID- 10476389 TI - Allyl chloride. PMID- 10476390 TI - Allyl isovalerate. PMID- 10476391 TI - 1,4-Benzoquinone (para-quinone). PMID- 10476392 TI - 1,4-Benzoquinone dioxime. PMID- 10476393 TI - Benzyl acetate. PMID- 10476394 TI - Bis(2-chloroethyl)ether. PMID- 10476395 TI - 1,2-Bis(chloromethoxy)ethane. PMID- 10476396 TI - 1,4-Bis(chloromethoxymethyl)benzene. PMID- 10476397 TI - Bis(2-chloro-1-methylethyl)ether. PMID- 10476398 TI - Bis(2,3-epoxycyclopentyl)ether. PMID- 10476399 TI - Bisphenol A diglycidyl ether. PMID- 10476400 TI - Bromochloroacetonitrile. PMID- 10476401 TI - Bromodichloromethane. PMID- 10476402 TI - Bromoethane. PMID- 10476403 TI - Bromoform. PMID- 10476404 TI - beta-Butyrolactone. PMID- 10476405 TI - Carbazole. PMID- 10476406 TI - Chloroacetonitrile. PMID- 10476407 TI - Chlorodibromomethane. PMID- 10476408 TI - Chlorodifluoromethane. PMID- 10476409 TI - Chloroethane. PMID- 10476410 TI - Chlorofluoromethane. PMID- 10476411 TI - 2-Chloro-1,1,1-trifluoroethane. PMID- 10476412 TI - Cyclohexanone. PMID- 10476413 TI - Decabromodiphenyl oxide. PMID- 10476414 TI - Dibromoacetonitrile. PMID- 10476415 TI - Dichloroacetonitrile. PMID- 10476416 TI - Dichloroacetylene. PMID- 10476417 TI - trans-1,4-dichlorobutene. PMID- 10476418 TI - 1,2-Dichloropropane. PMID- 10476419 TI - 1,2-Diethylhydrazine. PMID- 10476420 TI - Diethyl sulfate. PMID- 10476421 TI - Diglycidyl resorcinol ether. PMID- 10476422 TI - Diisopropyl sulfate. PMID- 10476423 TI - 1,1-Dimethylhydrazine. PMID- 10476424 TI - Dimethyl hydrogen phosphite. PMID- 10476425 TI - 3,4-Epoxy-6-methylcyclohexylmethyl 3,4-epoxy-6-methylcyclohexane carboxylate. PMID- 10476426 TI - cis-9,10-Epoxystearic acid. PMID- 10476427 TI - Ethyl acrylate. PMID- 10476428 TI - Glycidaldehyde. PMID- 10476430 TI - Isopropyl oils. PMID- 10476429 TI - Hexamethylphosphoramide. PMID- 10476431 TI - Lauroyl peroxide. PMID- 10476432 TI - Methyl acrylate. PMID- 10476433 TI - 2-Methylaziridine (propyleneimine). PMID- 10476434 TI - Methyl iodide. PMID- 10476435 TI - Morpholine. PMID- 10476436 TI - 1,5-Naphthalene diisocyanate. PMID- 10476437 TI - Pentachloroethane. PMID- 10476438 TI - Phenyl glycidyl ether. PMID- 10476439 TI - Tetrakis(hydroxymethyl)phosphonium salts. PMID- 10476440 TI - Trichloroacetonitrile. PMID- 10476441 TI - Triethylene glycol diglycidyl ether. PMID- 10476442 TI - Tris(2-chloroethyl) phosphate. PMID- 10476443 TI - 1,2,3-Tris(chloromethoxy)propane. PMID- 10476444 TI - Vinylidene fluoride. PMID- 10476445 TI - Acrylonitrile. PMID- 10476446 TI - 1,3-Butadiene. PMID- 10476447 TI - Chloroprene. PMID- 10476448 TI - Dichloromethane. PMID- 10476449 TI - Acetaldehyde. PMID- 10476450 TI - Aziridine. PMID- 10476451 TI - Benzoyl peroxide. PMID- 10476452 TI - n-Butyl acrylate. PMID- 10476453 TI - gamma-Butyrolactone. PMID- 10476454 TI - Caprolactam. PMID- 10476455 TI - Carbon tetrachloride. PMID- 10476456 TI - Catechol. PMID- 10476457 TI - alpha-Chlorinated toluenes and benzoyl chloride. PMID- 10476458 TI - 1,2-Dibromo-3-chloropropane. PMID- 10476459 TI - 1,2-Dichloroethane. PMID- 10476460 TI - Dimethylcarbamoyl chloride. PMID- 10476461 TI - Dimethylformamide. PMID- 10476462 TI - Dimethyl sulfate. PMID- 10476463 TI - 1,4-Dioxane. PMID- 10476464 TI - Epichlorohydrin. PMID- 10476465 TI - 1,2-Epoxybutane. PMID- 10476466 TI - Ethylene dibromide (1,2-dibromoethane). PMID- 10476467 TI - Hydrogen peroxide. PMID- 10476468 TI - Hydroquinone. PMID- 10476469 TI - Methyl bromide. PMID- 10476470 TI - Methyl chloride. PMID- 10476471 TI - Phenol. PMID- 10476472 TI - Polychlorophenols and their sodium salts. PMID- 10476473 TI - 1,1,2,2-Tetrachloroethane. PMID- 10476474 TI - Toluene. PMID- 10476475 TI - Toluene diisocyanates. PMID- 10476476 TI - 1,1,1-Trichloroethane. PMID- 10476477 TI - Tris(2,3-dibromopropyl) phosphate. PMID- 10476478 TI - Vinyl bromide. PMID- 10476479 TI - [Clinical diagnosis of bronchial asthma]. PMID- 10476480 TI - [Diagnostics in bronchial asthma]. PMID- 10476481 TI - [Bronchial asthma and chronic obstructive lung disease (COPD). Differences and similarities]. PMID- 10476482 TI - [Occupational and environmental bronchial asthma]. PMID- 10476483 TI - [Immunologic basis of bronchial asthma]. PMID- 10476484 TI - [Long-term treatment of bronchial asthma in adults]. PMID- 10476485 TI - [Disorders of sodium and water balance. Diagnosis and therapy]. PMID- 10476486 TI - [55-year old patient with fatigue, ascites, recurrent diarrhea and eosinophilia]. PMID- 10476487 TI - [38-year old woman with malaise and weight loss]. PMID- 10476488 TI - [On the contribution U.O. Wenzel, B.Pfalzer, I.Meinertz, R.A. Stahl (1999). A 73 year old woman with hypertensive crisis. Internist 40:205-9. Case report]. PMID- 10476489 TI - [Effects of excess iodine after the use of contrast media]. PMID- 10476490 TI - [Osteoporosis prophylaxis in men? Adverse effects of total androgen blockade in carcinoma of the prostate]. PMID- 10476491 TI - [Diagnosis of Clostridium difficile as cause of diarrhea]. PMID- 10476492 TI - [Antithrombotic therapy. Modern therapeutic approaches]. PMID- 10476493 TI - New enhancement procedures to optimize cataract and refractive surgery. PMID- 10476494 TI - Phacoemulsification after vitrectomy. PMID- 10476495 TI - Effect of lidocaine on photophobia. PMID- 10476496 TI - Preventing intraoperative ocular damage. PMID- 10476497 TI - Consultation section. Cataract surgical problem. PMID- 10476498 TI - Superficial lamellar keratectomy using an automated microkeratome to excise corneal scarring caused by photorefractive keratectomy. AB - Corneal haze is a recognized complication of excimer laser photorefractive keratectomy. When severe, it can result in the formation of a dense corneal scar, with a reduction in best corrected visual acuity. In extreme cases, medical treatment and repeated phototherapeutic excimer ablations may fail to restore corneal transparency. In these patients, penetrating keratoplasty (PKP) has been used to restore visual function. We describe a technique for excising a superficial corneal scar using an automated microkeratome. The resultant corneal surface is quite smooth, and good visual acuity may be restored without resorting to PKP. PMID- 10476499 TI - Suction posterior capsulorhexis. AB - A technique for creating a posterior capsulorhexis during phacoemulsification is presented. It can be used in cases with posterior capsule tears or opacities. The free edge of the capsule is grasped with suction using a 2 mL syringe and a 27 gauge Rycroft cannula introduced via the paracentesis. The edge is then manipulated to produce a continuous curvilinear opening in the posterior capsule. The combination of a closed eye plus the use of a viscoelastic agent in the anterior chamber and capsular bag minimizes the possibility of vitreous prolapse during the maneuver. Occlusion of the cannula tip by the posterior capsule reduces the risk of vitreous aspiration. In-the-bag intraocular lens implantation is readily achieved. PMID- 10476500 TI - Excimer ablation design and elliptical transition zones. AB - PURPOSE: To describe the rationale behind elliptical and other transition designs used with the excimer laser. SETTING: Casey Eye Institute, Oregon Health Sciences University, Portland, Oregon, USA. METHODS: Ablation zone designs were analyzed for the number of transition points for myopia and hyperopia. The advantages and disadvantages of elliptical transition zones are demonstrated graphically, with an emphasis on smooth ablation zone design to maximize the optics and biologic tolerance by the eye. RESULTS: The use of an individualized elliptical transition maximizes a circular effective optical zone and can enhance the smoothness of the transition zone while minimizing excessive tissue removal. CONCLUSION: Elliptical transition zones may improve the optics and biologic tolerance of excimer laser treatments. PMID- 10476501 TI - Results of laser in situ keratomileusis in hyperopic compound astigmatism. AB - PURPOSE: To evaluate the efficacy and safety of steepening the flatter meridian with laser in situ keratomileusis (LASIK) to correct hyperopic compound astigmatism using the Schwind-Keratom laser with the MultiScan System and the active tracking system for centering the ablation. SETTING: Instituto Barraquer de America, Bogota, Colombia. METHODS: From June to December 1997, LASIK was performed in 111 eyes to correct congenital hyperopic compound astigmatism. Preoperatively, the mean spherical equivalent was +2.58 diopters (D) (range +1.50 to +3.50 D) in 37 eyes (low dioptric group), with a mean sphere of +3.66 D (range +1.75 to +5.50 D) and a mean cylinder of -2.11 D (range -0.50 to -5.00 D); +4.71 D (range +3.51 to +6.00 D) in 50 eyes (middle dioptric group), with a mean sphere of +5.58 D (range +4.00 to +7.00 D) and a mean cylinder of -1.83 D (range -0.50 to -4.50 D); and +7.26 D (range +6.01 to +10.00 D) in 24 eyes (high dioptric group), with a mean sphere of +8.25 D (range +6.50 to +10.00 D) and a mean cylinder of -1.98 D (range -0.50 to -4.00 D) in 24 eyes. Postoperative results were measured at 10 days and 6 months. RESULTS: At 6 months, the mean residual subjective spherical defect was +0.32 D (range -0.50 to +1.00 D) in the low dioptric group, with a mean cylinder of -0.61 D (range 0.00 to -1.75 D); all eyes were within +/- 1.00 D of emmetropia. The mean defect was +0.82 D (range 0.00 to +2.50 D) in the middle dioptric group, with a mean cylinder of -0.61 D (range 0.00 to -2.50 D); 80% of eyes were within +/- 1.00 D of emmetropia. The mean defect was +1.10 D (range -0.50 to +3.00 D) in the high dioptric group, with a mean cylinder of -1.39 D (range 0.00 to -3.25 D); 77% of eyes were within +/- 1.00 D of emmetropia. In the study group as a whole, 90% of eyes had a cylinder correction within +/- 1.00 D of emmetropia. Uncorrected visual acuity (UCVA) was 0.50 (20/40) or better in 23.4% of eyes preoperatively and 0.50 (20/40) or better in 71.0% 6 months postoperatively. CONCLUSIONS: Steepening the flatter meridian with the MultiScan System safely and effectively corrected hyperopic astigmatism. The rapid recovery of UCVA was due not only to correction of the ametropia, but also to the centering provided by the active tracking system. PMID- 10476502 TI - Minus-power intraocular lenses to correct refractive errors in myopic pseudophakia. AB - PURPOSE: To evaluate the effectiveness of a secondary, piggyback, minus-power intraocular lens (IOL) to correct the refractive error in patients with myopic pseudophakia. METHODS: In this prospective noncomparative cohort study, 51 myopic pseudophakic patients received implantation of a minus-power IOL as a secondary procedure to correct residual pseudophakic myopia. RESULTS: The mean residual myopia of -3.05 diopters (D) was reduced to -0.38 D. All eyes were within +/- 1.00 D of the desired refraction. Uncorrected visual acuity was 20/40 or better in 72% of eyes, and best corrected visual acuity was 20/40 or better in 96%. Uncorrected visual acuity improved by 2 or more lines in 85% of eyes and by 5 or more lines in 65%. CONCLUSION: Clinical outcomes can now be improved in patients with myopic pseudophakia whose previous options (i.e., lens exchange or refractive surgery) were more traumatic or less predictable. PMID- 10476503 TI - Effect of age at time of cataract surgery on subsequent axial length growth in infant eyes. AB - PURPOSE: To determine whether removal of the crystalline lens and placement of an intraocular lens (IOL) in human infant eyes retard the growth of the pseudophakic eye. METHODS: A unilateral lensectomy with placement of a posterior chamber IOL in the sulcus was performed in 11 infants between 2 and 4 months of age. Axial length measurements of both eyes were obtained preoperatively and postoperatively. RESULTS: Patients were followed for a mean of 5.6 years. In 7 patients, the mean axial growth was 0.46 mm less in the pseudophakic eye than in the fellow eye (range 0.15 to 0.70 mm). In 1 patient, there was no interocular axial length difference and in 3, the pseudophakic eye was longer. When measurements from the only patient with microphthalmia were excluded, the interocular difference in axial growth was highly significant (signed rank test, P = .006). Median visual acuity of the pseudophakic eyes at the last follow-up was 20/60 (range 20/30 to 20/200). The final visual acuity in the pseudophakic eyes did not correlate with the degree of interocular axial length difference (P > .05). CONCLUSIONS: Our study suggests there may be a reduction in axial growth in infantile eyes following cataract extraction and IOL implantation. This effect probably reduces the magnitude of the myopic shift in these eyes. PMID- 10476504 TI - Combined cataract and glaucoma surgery: trabeculectomy versus endoscopic laser cycloablation. AB - PURPOSE: To determine whether combined cataract surgery with endoscopic laser cycloablation produces less inflammation than cataract surgery combined with a filtering procedure. SETTING: Taylor Regional Hospital (surgeries) and EyeSight Associates (examinations), Warner Robins, Georgia, USA. METHODS: A randomized prospective study was conducted of 58 eyes of 58 patients comparing endoscopic laser cycloablation performed through a cataract incision at the time of cataract surgery with combined trabeculectomy and cataract surgery. RESULTS: Mean follow up was 2 years. At the final available visit, 30% of endoscopic laser patients achieved intraocular pressure control (below 19 mm Hg) without medication and 65% with medication. Forty percent of trabeculectomy patients achieved control without medication and 52% with medication. Four endoscopic laser patients (14%) and 3 trabeculectomy patients (10%) were considered treatment failures (required additional surgical intervention). CONCLUSION: Endoscopic laser cycloablation performed through a cataract incision was a reasonably safe and effective alternative to combined cataract and trabeculectomy surgery, providing an option for cataract patients who have glaucoma requiring surgical intervention. PMID- 10476505 TI - Late pigmented-membrane formation on silicone intraocular lenses after phacoemulsification with or without trabeculectomy. AB - PURPOSE: To determine the incidence of late inflammatory membrane formation with pigment precipitates on foldable silicone or poly(methyl methacrylate) (PMMA) intraocular lenses (IOLs) after phacoemulsification with or without simultaneous trabeculectomy and to identify probable causative factors. SETTING: Eye Department, Bnai-Zion Medical Center, and the Faculty of Medicine, the Technion, Haifa, Israel. METHODS: In this retrospective study, 155 eyes (140 patients) were divided into 4 groups by the surgery performed: Group 1, phacoemulsification combined with trabeculectomy and foldable silicone IOL implantation (12 eyes); Group 2, phacoemulsification combined with trabeculectomy and PMMA IOL implantation (15 eyes); Group 3, phacoemulsification alone with foldable silicone IOL implantation (66 eyes); Group 4, phacoemulsification alone with PMMA IOL implantation (62 eyes). Preoperative, intraoperative, and postoperative data were compared. RESULTS: Late inflammatory membranes were found on the anterior surface of 33% of the IOLs in Group 1, 3% in Group 3, and none in Groups 2 and 4. Membranes developed 3 to 4 months after surgery, were resistant to topical corticosteroid treatment and required repeated neodymium:YAG laser treatments. No correlation with preoperative, intraoperative, or postoperative factors was found. CONCLUSION: Foldable silicone IOLs may induce late postoperative inflammatory membranes with pigment precipitates, especially after combined phacoemulsification and trabeculectomy. PMID- 10476506 TI - Functional improvement after phacoemulsification cataract surgery. AB - PURPOSE: To evaluate subjective functional visual improvement after phacoemulsification and intraocular lens (IOL) implantation and identify patient characteristics and treatment features associated with no functional improvement. SETTING: Academic referral practice, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas, USA. METHODS: A retrospective review was performed of the surgical results in 106 consecutive eyes of 102 patients who completed the American Society of Cataract and Refractive Surgery (ASCRS) Cataract Data Collection Form preoperatively and 1 month postoperatively. One surgeon performed all the surgery. The relationship of patient characteristics and treatment features and reported visual function was analyzed using an analysis of variance and the Wilcoxon rank sum and Fisher exact tests. RESULTS: One month postoperatively, all patients demonstrated improved visual acuity. However, 15 patients (14%) reported unimproved or decreased visual function. No statistically significant intergroup difference (P > .05) was detected in mean preoperative, postoperative, and fellow eye best spectacle corrected visual acuities or refractive errors; reported severity of preoperative symptoms or satisfaction with vision; reported preoperative daily activity functioning or prevalence of hypertension and diabetes; type of anesthesia, lens material, or wound closure used; intraoperative or postoperative complication rates. However, patients reporting unimproved vision tended to be older and less satisfied with their quality of life and medical care than those reporting improved vision (P = .02, .03, .01, respectively). CONCLUSIONS: Self-reported visual function generally improved after phacoemulsification and IOL implantation; however, some older patients less satisfied with their quality of life and medical care reported no functional improvement despite improved visual acuity. These patients might benefit from an extended preoperative discussion of postoperative expectations. PMID- 10476507 TI - Comparison of 4 topical anesthetic agents for effect and corneal toxicity in rabbits. AB - PURPOSE: To compare the onset time, duration of action, corneal toxicity, and corneal epithelial healing time of 4 topical anesthetic agents in rabbits. SETTING: Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada. METHOD: Fifty-six rabbits were treated with 4 topical anesthetics (bupivacaine, lidocaine, procaine, and benzocaine) at different concentrations and different pH of solutions. Corneal sensation, corneal toxicity, and corneal epithelial healing time were measured. RESULTS: The onset time of all 4 anesthetic agents was within 1 minute; however, bupivacaine and lidocaine produced significantly longer action than procaine or benzocaine (P < .05). Buffered bupivacaine and lidocaine had a significantly longer anesthetic effect than that of the nonbuffered solutions (P < .05). No significant effect on corneal epithelial healing time or corneal toxicity was observed. CONCLUSION: Topical bupivacaine and lidocaine had a longer anesthetic effect, particularly in buffered solutions. No significant corneal toxicity was observed. PMID- 10476508 TI - Scleral perforation during retrobulbar and peribulbar anesthesia: risk factors and outcome in 50,000 consecutive injections. AB - PURPOSE: To measure the frequency of scleral perforation and identify related risk factors during local anesthetic injection for intraocular surgery. SETTING: Multispecialty eye hospital. METHODS: All patients (n = 50,000) having retrobulbar (26,857) or peribulbar (23,143) injections at the King Khaled Eye Specialist Hospital were reviewed. Cases of scleral perforation were analyzed for potential technical and ophthalmic risk factors, management of injuries, and visual and anatomic outcomes. Mean follow-up was 14.4 months (range 8 to 24 months). RESULTS: Seven (0.014%) needlestick injuries were identified, all of which had posterior staphyloma as the only identifiable risk factor. Applying a previously measured prevalence of 10.7% for posterior staphyloma in our surgery patients gave a scleral perforation rate of 0.13% (7 of 5350) for staphylomatous eyes. All perforated globes had originally planned cataract extraction within 8 weeks of injury. Additional management consisted of observation (2 cases), cryotherapy (2 cases), and vitreoretinal procedures for retinal detachment (3 cases) and subretinal hemorrhage (1 case). At last follow-up, all retinas were attached and 3 cases (42.8%) had a visual acuity of worse than 20/160. Both cases requiring multiple retinal detachment surgeries developed proliferative vitreoretinopathy and poor visual acuity. CONCLUSIONS: Eyes with posterior staphyloma sustained needlestick injuries at a rate of 1 in 760 compared with 0 injection perforations in more than 44,000 nonstaphylomatous eyes. PMID- 10476509 TI - Cluster of diplopia cases after periocular anesthesia without hyaluronidase. AB - PURPOSE: To describe a cluster of cases of iatrogenic diplopia after cataract surgery that occurred in 1998, when hyaluronidase was unavailable for use in periocular anesthetic regimens. SETTING: The clinical practices of the authors. METHODS: This study comprised a retrospective chart review. RESULTS: Twenty-five cases of transient or permanent diplopia were reported. Of these, 13 eyes had retrobulbar and 10 had peribulbar injections; in 2 cases the injection technique was unknown. The inferior rectus was affected in 19 eyes; of these, 1 had a temporary palsy and 18 had permanent restriction. Temporary paresis developed in the lateral rectus in 5 cases and the superior rectus in 2. Eleven cases were submitted by 4 anterior segment surgeons, who collectively had a zero incidence of iatrogenic postoperative diplopia in the preceding 4 to 11 years of practice (approximately 6900 cases). CONCLUSION: Hyaluronidase may be more important than previously suspected in preventing anesthetic-related damage to the extraocular muscles. The inferior rectus muscle is particularly vulnerable, presumably because of the injection technique. PMID- 10476510 TI - Anesthetic dose and analgesic effects of sub-Tenon's anesthesia in cataract surgery. AB - PURPOSE: To compare the analgesic effects of different doses of sub-Tenon's anesthesia in cataract surgery by assessing patient response to visceral stimulus. SETTING: Inouye Eye Hospital, Tokyo, Japan. METHODS: A prospective study was done of 1019 eyes of 1019 patients having phacoemulsification and posterior chamber intraocular lens implantation. They received a 1.0 mL (391 eyes), 2.0 mL (366 eyes), or 3.0 mL (262 eyes) anesthetic infiltration into the sub-Tenon's space. Pain scores were recorded when the anterior chamber was irrigated with an acetylcholine chloride solution to attain miosis after lens implantation. RESULTS: The distribution of pain scores was significantly different among the 3 groups (P < .0001, Kruskal-Wallis test). Multiple comparison revealed that the 3.0 mL anesthetic infiltration offered significantly higher analgesic effects than the 2 lower doses. The 3.0 mL sub-Tenon's anesthesia effectively blocked the visceral stimulus. CONCLUSION: For cataract surgery, 3 mL is the optimal dose of anesthetic solution in sub-Tenon's anesthesia. PMID- 10476511 TI - Driving simulation study: bilateral array multifocal versus bilateral AMO monofocal intraocular lenses. AB - PURPOSE: To determine whether differences exist in the driving performance of patients with bilateral Array multifocal intraocular lenses (IOLs) and those with bilateral AMO monofocal IOLs under low-contrast environmental conditions. SETTING: The Iowa Driving Simulator at the Center for Computer Aided Design, the University of Iowa, Iowa City, Iowa, USA. METHODS: This prospective study was a test-operator-masked, parallel-group comparison of the driving performance of 33 bilateral multifocal IOL patients and 33 bilateral monofocal IOL patients from the U.S. Array Multifocal study. Driving performance was evaluated under 3 poor visibility conditions (clear weather at night, clear weather at night in the presence of a glare source, and fog). Measures of performance included recognition rates and distances for signs, as well as detection rates, distances, and avoidance behaviors for hazards. Contrast acuity and sensitivity were also measured to evaluate possible correlations with driving performance. RESULTS: No statistically significant differences between the IOL groups were found in 26 of 30 comparisons (86.7%). The monofocal group performed better than the multifocal group in comparisons in which there were statistically significant differences: the percentage of correctly recognized warning signs at night in clear weather (P = .028), sign recognition distances for guide (P = .030) and warning (P = .036) signs in fog, and the detection distance for 1 of 4 hazards (suitcase; P = .026). Correlation coefficients between driving performance and low-contrast acuity and sensitivity were statistically significant; however, they were low and not likely predictive of driving performance. CONCLUSION: Differences between patients with bilateral multifocal IOLs and those with bilateral monofocal IOLs were detected; however, the results indicate no consistent difference in driving performance and safety. PMID- 10476512 TI - Long-term visual results of bifocal intraocular lens implantation. AB - PURPOSE: To evaluate the long-term visual results and variability in contrast sensitivity in patients with heparin-surface-modified (HSM) diffractive bifocal intraocular lenses (IOLs). SETTING: Institute of Ophthalmology, University of Catania, Italy. METHODS: In this prospective study, visual acuity (distance and near) and contrast sensitivity were measured in 35 patients who had phacoemulsification with bifocal diffractive IOL (model 811E, Pharmacia) implantation. Patient satisfaction was also evaluated using a questionnaire. Mean follow-up was 20 months (range 18 to 24 months). RESULTS: At the last examination, mean distance visual acuity was 0.79 +/- 0.2 (SD) without correction and 1.0 +/- 0.15 with best correction. Mean uncorrected near visual acuity was J1.6 +/- J0.77 and with best distance correction, J1.19 +/- J0.49. No statistically significant changes in visual acuity were evident at the last follow-up (Student t test). No changes were found in contrast sensitivity reduction over time, nor were late postoperative complications noted. Overall patient satisfaction was rated as good by 94.3% of patients with best distance correction. CONCLUSIONS: The diffractive bifocal HSM IOL provided good visual performance for distance and near over time. In relation to the low rate of postoperative complications, the slight contrast sensitivity reduction was stable during follow-up. PMID- 10476513 TI - Material properties of ophthalmic sutures after sterilization and disinfection. AB - PURPOSE: To assess the effects that sterilization and disinfection have on the tensile properties of commonly used ophthalmic sutures. SETTING: University of Bristol, Bristol, United Kingdom. METHODS: The sutures 10-0 nylon, 10-0 polypropylene (Prolene), 8-0 virgin silk, and 8-0 polyglactin 910 (Vicryl) were subjected to thermal or chemical sterilization/disinfection including autoclaving, boiling, and immersion in alcohol or glutaraldehyde. Measurements of tensile strength, suture extension to the point of failure, and suture stiffness were then performed. RESULTS: Nylon and Prolene were little affected by all sterilization and disinfection methods. Virgin silk and Vicryl were weakened by thermal methods of sterilization and disinfection. Both Vicryl, and to a lesser extent virgin silk, appeared to be strengthened by immersion in alcohol or glutaraldehyde. CONCLUSION: The casual reuse of ophthalmic sutures is not recommended. However, when there is no alternative, nylon and Prolene monofilaments can be autoclaved or immersed in fresh activated glutaraldehyde, and Vicryl and virgin silk can be immersed in fresh activated glutaraldehyde without significant loss of tensile strength. PMID- 10476514 TI - Extracapsular cataract extraction with a sutureless incision for dense cataracts. AB - PURPOSE: To evaluate the use of extracapsular cataract extraction (ECCE) via a sutureless incision for dense cataracts. SETTING: Nagata Eye Clinic, Nara, Japan. METHODS: This retrospective study comprised 51 eyes of 45 consecutive patients with dense cataracts who had ECCE with a sutureless incision between January 1996 and April 1998. A scleral incision from 6.0 to 8.5 mm was made at 12 o'clock or between 9 and 12 o'clock (oblique incision). Measures of outcome included postoperative visual acuity, surgically induced astigmatism (polar value method and vector analysis), complications, and changes in corneal endothelial cell density and morphology. RESULTS: Self-sealing was achieved in 45 eyes (88.2%), but additional sutures were required in 6 (11.8%). Intraoperative complications included posterior capsule rupture in 3 eyes (5.9%) and iris prolapse in 2 (3.9%). Corneal flattening against the preoperative steep meridian was observed in the 12 o'clock incision group (0.24 diopter [D] +/- 1.23 [SD]) and in the oblique incision group (0.17 +/- 0.89 D). By vector analysis, the surgically induced vector was 1.41 +/- 0.72 D in the 12 o'clock incision group and 1.02 +/- 0.66 D in the oblique incision group. After surgery, the mean cell loss was 8.2% +/- 12.5%. There were no significant differences, however, between other preoperative and postoperative morphometric indexes. CONCLUSION: This fast, safe, and inexpensive technique may be a viable treatment for dense cataracts with large, hard nuclei. PMID- 10476515 TI - Postoperative inflammation: extracapsular cataract extraction versus phacoemulsification. AB - PURPOSE: To quantitatively compare postoperative inflammation after extracapsular cataract extraction (ECCE) with that after phacoemulsification in an Asian population. SETTING: Singapore National Eye Center, Singapore. METHODS: In this prospective, randomized, double-masked clinical trial, patients having cataract surgery were randomized to receive ECCE (n = 16) or phacoemulsification (n = 18). Diabetics were excluded. Two surgeons performed both types of surgery and implanted a 6.0 mm optic intraocular lens. Inflammation was assessed qualitatively by slitlamp grading of cells and flare and quantitatively using the Kowa flare meter. One independent postoperative investigator performed the slitlamp examination and laser flare meter readings. RESULTS: The ECCE and phacoemulsification groups were comparable (P > .05) in age, sex, ethnicity, and preoperative flare levels. The combined slitlamp inflammatory scores (anterior chamber cells and flare) and mean laser flare meter readings showed the ECCE group had significantly higher mean flare measurements than the phacoemulsification group at days 4 (P = .0012), 8 (P = .0013), 15 (P = .0013), 30 (P = .0004), and 60 (P = .0164). Flare levels in the ECCE group returned to preoperative values by the second month; the phacoemulsification group achieved preoperative levels by 1 month. The clinical inflammatory assessment score correlated closely to the flare level readings. CONCLUSION: Phacoemulsification induced less inflammation than ECCE, with the difference most marked in the first month after surgery. PMID- 10476516 TI - Pseudophakic cystoid macular edema treated with high-dose intravenous methylprednisolone. AB - PURPOSE: To evaluate the efficacy of high-dose methylprednisolone in the treatment of pseudophakic cystoid macular edema (CME). SETTING: University affiliated hospital. METHODS: Four patients with pseudophakic CME who reported decreased visual acuities were treated with oral prednisolone (20 mg daily for 5 days), oral acetazolamide (500 mg daily for 5 days), and topical dexamethasone 0.1% and diclofenac 0.1% (4 times a day for 14 days). The CME did not resolve. Next, they were treated with high-dose (1000 mg daily) intravenous methylprednisolone for 3 days. RESULTS: In 3 of 4 eyes, the CME resolved and visual acuity improved. CONCLUSION: High-dose methylprednisolone may be effective in the treatment of pseudophakic CME. PMID- 10476517 TI - Experimental staining of the anterior lens capsule in albino rabbits. AB - PURPOSE: To study the value and safety of staining the anterior lens capsule in albino rabbits. SETTING: Ophthalmology Department, Cairo University, and Ophthalmic Pathology Department, Research Institute of Ophthalmology, Cairo, Egypt. METHODS: The experiment was divided into 3 stages. First, the capacity of different concentrations of crystal violet solution to stain the anterior lens capsule of postmortem albino rabbit eyes was tested. The toxicity of different concentrations of the dye (2% to 0.25%) was then tested to determine the highest concentration that was nontoxic to the cornea and trabecular meshwork. The third step was to detect possible toxicity of lower concentrations (0.1% to 0.05%). Different dye concentrations were injected into the anterior chamber of the rabbit eyes. The eyes were examined after 1 and 3 days, 1 week, and 1 month using light microscopy and scanning and transmission electron microscopy. RESULTS: Different concentrations of crystal violet stained the anterior lens capsule, allowing for easy capsulorhexis. The use of the 2% and 1% concentrations was accompanied with irreversible damage to all corneal layers. The use of the 0.5% concentration caused damage to stromal keratocytes and endothelium. The use of the 0.25% concentration did not damage any corneal layer or the trabecular meshwork. Lower concentrations of 0.1% and 0.05% also stained the capsule, providing good visibility for successful capsulorhexis, and were less toxic to the corneal endothelium. CONCLUSION: Staining the anterior lens capsule with 0.25% to 0.05% concentrations of crystal violet solution caused no injury to the cornea or trabecular meshwork in albino rabbit eyes. PMID- 10476518 TI - Endophthalmitis after phakic intraocular lens implantation to correct high myopia. AB - A 36-year-old woman had uneventful implantation of an angle-supported anterior chamber phakic intraocular lens (IOL) to correct high myopia. On the first postoperative day, she developed infectious endophthalmitis likely associated with intraoperative contamination. Group B beta-hemolytic Streptococcus endophthalmitis was confirmed by microbiologic studies. To our knowledge, this is the first documented case of infectious endophthalmitis after anterior chamber phakic IOL implantation to correct high myopia. PMID- 10476519 TI - Treatment of traumatic cyclodialysis with vitrectomy, cryotherapy, and gas endotamponade. AB - An aphakic patient with severe chronic hypotony had an alternative treatment of a traumatic cyclodialysis cleft: a 3-port pars plana vitrectomy, cryotherapy of the cleft, and fluid-gas exchange with subsequent supine positioning. The therapeutic principle was mechanical apposition of the detached ciliary muscle to the scleral spur by the gas bubble and scar induction by cryotherapy. Intraocular pressure increased to within normal ranges, and visual acuity improved over a 15 month follow-up. PMID- 10476520 TI - Traumatic cataract presenting with unilateral nasal hemianopsia. AB - A 56-year-old man developed a nasal field defect in his left eye 3 months after a traumatic accident. An examination showed a posterior subcapsular cataract in the left eye with no neurologic deficits. Humphrey 24-2 visual field testing revealed a nasal hemianopsia in the left eye. After cataract extraction and intraocular lens implantation, the patient's visual field returned to normal. This case shows that a cataract can present with a localized visual field deficit, which may be corrected by cataract extraction. PMID- 10476521 TI - What is your diagnosis? Tracheal rupture. PMID- 10476522 TI - Antihistamines in the management of allergic pruritus in dogs and cats. AB - Antihistamines clearly have a place in the management of pruritus in the atopic dog and cat and, in this paper, important aspects of pharmacology and pharmacokinetics, adverse effects and precautions, and clinical use of these compounds are reviewed. Successful use of antihistamines is dependent on, among other considerations, the adherence to recommended doses and frequencies of administration, the recognition and control of concurrent and secondary factors, the willingness to try several different compounds, and the realisation that these agents are best employed in a preventive fashion. PMID- 10476523 TI - Measurement of titres of naturally occurring alloantibodies against feline blood group antigens in the UK. AB - Measurement of anti-A and anti-B haemagglutinating alloantibody titres was performed on serum samples from both pedigree (n = 61) and non-pedigree (n = 43) cats previously typed using a commercial blood typing kit. All 40 type B cats had anti-A antibody with titres ranging from 4 to 1600. Among the 61 type A sera tested, an anti-B agglutination titre of greater than 2 was recorded in 16.4 per cent. There was no significant association between serum alloantibody titre and cat breed or gender (P > 0.05). PMID- 10476524 TI - Observations on the development of congestive heart failure in Irish wolfhounds with dilated cardiomyopathy. AB - Records of previous electrocardiographic (ECG) and echocardiographic examinations of 39 Irish wolfhounds with cardiac failure, obtained during a 10-year survey, were examined to establish whether previously detected abnormalities might be prognostic indicators for the development of congestive heart failure (CHF) due to dilated cardiomyopathy (DCM) in later life. Eighteen dogs (46 per cent) had had atrial fibrillation (AF) at their first examination. Other ECG abnormalities detected at previous examinations were ventricular and supraventricular premature contractions, first and second degree atrioventricular block, P mitrale and left anterior fascicular and right bundle branch blocks. All 39 dogs had developed AF by the time of onset of CHF. The mean age at which CHF was diagnosed was 77 months in males and 86 months in females. The mean age at which AF was first detected was 45 months in males and 59 months in females, and the mean time from the first detection of AF to CHF was 27 months in males and 24 months in females. Previous echocardiographic examinations in eight dogs, over a period of up to five years, revealed progressive left atrial and left ventricular dilatation. Fractional shortening was reduced to below the normal range at the onset of CHF compared with previous examinations in three cases but increased in five. It appears, therefore, that certain ECG abnormalities, especially AF, and/or progressive ventricular and atrial dilatation, may be indicators of 'occult' DCM in wolfhounds. PMID- 10476525 TI - Free skin grafting for treatment of distal limb skin defects in cats. AB - The technique and results of free skin grafts have been described in dogs, horses, rabbits, goats and mice. The procedure in cats is, however, described only indirectly in papers relating to dogs. A standard technique has been developed by the authors for use in cats, and is reported for 17 grafts in 16 cats with traumatic injury to the legs resulting in large areas of skin loss. This paper describes the preparation of the wound for grafting, the harvesting of the graft, graft placement, postoperative care and the results of the application of this technique. The success rate in this series of cases was high. This was attributed to proper preparation of the recipient site, collection of the graft and postoperative bandaging. The results suggest that the success rate of free skin grafts in cats is considerably higher than that achieved by the present authors in dogs, and reported for dogs by other surgeons in the literature. PMID- 10476526 TI - Pericardial mesothelioma in a dog: long-term survival after pericardiectomy in combination with chemotherapy. AB - A six-year-old male crossbred dog was presented with clinical signs of right sided heart failure. Echocardiography demonstrated a pericardial effusion with cardiac tamponade, while pericardiocentesis and cytology did not reveal any evidence of malignancy. Pericardial drainage was performed twice over a period of three months to resolve haemodynamic impairment before a subtotal pericardiectomy was performed. Biopsy of parietal and visceral pericardium confirmed the diagnosis of pericardial mesothelioma. Intrathoracic cisplatin combined with intravenous doxorubicin were administered, although neutropenia, mild azotaemia and alopecia were noted as adverse reactions to these drugs. Intravenous cisplatin was repeated 45 days later after the signs of nephrotoxicity had resolved. The dog was still free of disease after 27 months. Intrathoracic chemotherapy after pericardiectomy and early diagnosis are recommended to improve prognosis, having achieved long-term survival in the present case. PMID- 10476527 TI - Intra-abdominal cryptococcosis in two dogs. AB - Intra-abdominal cryptococcosis was diagnosed in two young dogs. The first, an entire male border collie, was presented with vomiting. An abdominal mass detected during physical examination proved to be cryptococcal mesenteric lymphadenitis on exploratory laparotomy. The second dog, a female neutered giant schnauzer, was presented with neurological signs suggestive of encephalopathy. Intestinal cryptococcal granulomas were detected in an extensive diagnostic investigation which included abdominal ultrasonography. The gastrointestinal tract was considered the most likely portal of entry for cryptococcal organisms in both cases. Both dogs were treated using surgery and multiagent antifungal chemotherapy. The first case succumbed despite therapy, while the second dog was treated successfully as gauged by return to clinical normality and a substantial decline in the cryptococcal antigen titre which continued to fall after cessation of treatment. PMID- 10476528 TI - Transient diabetes insipidus in a dog with acromegaly. AB - A nine-year-old female beagle with acromegaly and extreme polyuria and polydipsia during dioestrus is described. It was demonstrated that polyuria was related to an inadequate rise of arginine-vasopressin (AVP) levels after water deprivation and stimulation with hypertonic saline. Administration of AVP did not lead to a significant increase in urine osmolality or reduction of urine volume. Clinical signs, except for bony changes, completely disappeared following ovariohysterectomy. PMID- 10476529 TI - An overall win on machines. PMID- 10476530 TI - The nurse as a moral agent in modern health care. PMID- 10476531 TI - Virginia M. Ohlson: international icon in public health nursing. Interview by Shirley H. Fondiller. PMID- 10476532 TI - Breakthroughs in nursing education: looking back, looking forward. PMID- 10476533 TI - Creating community-based care for the new millennium. PMID- 10476534 TI - Harnessing innovative technologies: what can you do with a shoe? PMID- 10476535 TI - Disseminating our breakthroughs: enacting a strategic framework. PMID- 10476536 TI - Suffer--one definition with commentary. PMID- 10476537 TI - Nursing research and the erosion of care. PMID- 10476538 TI - Lung cancer highlights. PMID- 10476539 TI - Hematologic malignancies. PMID- 10476540 TI - Progress in gynecologic oncology. PMID- 10476541 TI - Breast cancer highlights. PMID- 10476542 TI - Cancers in children infected with the human immunodeficiency virus. AB - The AIDS epidemic continues unabated in Africa, Asia, and South America, and since patients survive longer, the number of chronically immunocompromised individuals is increasing in Europe and the United States. The number of children with HIV infection who will ultimately develop a malignancy is not known. Currently, tumors represent about 2% of the AIDS-defining events in children in the United States, but the incidence might be different in developing countries. The most common tumors in HIV-infected children are non-Hodgkin's lymphoma, smooth muscle tumors (leiomyosarcomas), and Kaposi's sarcoma (only in Africa). This article provides an overview of epidemiology and clinical and pathological presentations, as well as preliminary data regarding treatment options in children with HIV-associated malignancies. PMID- 10476543 TI - A retrospective review of blood transfusions in cancer patients with anemia. AB - BACKGROUND: The factors contributing to blood transfusions in patients with anemia of chronic disease are not well documented in the literature. We analyzed all blood transfusion events within a single oncology practice to determine if certain chemotherapy drugs, cancer types, or other factors necessitated more frequent transfusions. PATIENTS AND METHODS: Out of 331 patients receiving chemotherapy, 103 (31%) patients received a blood transfusion in 1995. Each of these charts was reviewed and sorted by diagnosis, treatment medications, and past transfusion and/or treatment history. Hemoglobin levels were obtained for each transfusion received in 1995. RESULTS: The average hemoglobin at time of transfusion was 7.9 g/dl. Higher hemoglobin levels at transfusion were observed for patients over the age of 60 and patients who received prior chemotherapy. Lower hemoglobin levels at transfusion were observed for patients receiving Epoetin Alfa and sarcoma patients. The average number of red blood cell (RBC) units transfused in 1995 was 5.1 per patient. More units were given to patients receiving etoposide, while fewer units were given to those receiving ifosfamide. We created a transfusion severity index (TSI) to jointly measure these two variables. CONCLUSION: The results of this study identify transfusion needs associated with certain groups of cancer patients and with certain types of chemotherapy drugs. PMID- 10476544 TI - Race, ethnicity, and the patient-caregiver relationship. AB - Shortly before his death in 1995, Kenneth B. Schwartz, a cancer patient at Massachusetts General Hospital (MGH), founded The Kenneth B. Schwartz Center at MGH. The Schwartz Center is a non-profit organization dedicated to supporting and advancing compassionate health care delivery, which provides hope to the patient, gives support to caregivers, and encourages the healing process. The center sponsors the Schwartz Center Rounds, a monthly multidisciplinary forum where caregivers reflect on important psychosocial issues faced by patients, their families, and their caregivers, and gain insight and support from fellow staff members. Racial discrimination is a pervasive problem with multiple damaging effects. It is naive to believe that medicine is somehow immune to race-based practices, but there is a growing literature detailing poorer disease-specific outcomes in minority populations for a range of illnesses. A recent study in the New England Journal of Medicine has implicated physician prejudice as a significant contributing factor. The March 1999 Schwartz Center Rounds sought to explore the influence of ethnic bias on the patient-provider interaction and the quality of health care delivery. Using a different format with a current affairs video clip and an interactive panel discussion, participants were encouraged to identify the often subconscious racial prejudices which may undermine their relationships with patients. Staff members were challenged to think creatively about how institutions and individuals might promote "cultural competence" and a more equitable health care environment. PMID- 10476545 TI - Apoptosis. AB - Mechanisms in Hematology is a book with an accompanying interactive CD-ROM designed to assemble basic concepts that underlie clinical understanding and progress. It is presented as a concise text with a series of diagrams that distill diffuse information into a compact form. The interactive CD, in particular, brings many of the processes "to life" as details of the more complex pathways are conveyed in clear visual images. The text begins with the basic molecular biology that underlies hematological and oncological physiology/pathology--cell signaling, adhesion molecules, and apoptosis. This is followed by sections, among others, on hematopoiesis, iron, B12, and folate metabolism, neutrophil function, immunoproteins, chemotherapy, and coagulation. With the permission of the authors and publisher. The Oncologist has reproduced the section on apoptosis, which we think our readers will enjoy. PMID- 10476546 TI - The molecular perspective: methotrexate. PMID- 10476547 TI - Tsetse and trypanosomiasis control problems in south-east Uganda: past, present and alternative strategies. AB - Tsetse and trypanosomiasis in South-East Uganda are reviewed. This paper examines why in nearly 100 years no appreciable progress has been made in tsetse and trypanosomiasis control. It points out that tsetse control strategies in the past have relied on sophisticated technologies such as aerial spraying, which are inappropriate to Uganda's economic and environmental situation. With the vector, G. f. fuscipes, being peridomestic and the transmission cycle undoubtedly domestic animal-fly-man, community participation using appropriate technologies such as low-cost traps/targets and integrating farming activity with tsetse control seem to be the most appropriate approach in South-East Uganda. Savings in expenditure on vector control are discussed in light of diminishing resources. PMID- 10476548 TI - Shared vector-borne zoonoses of the Old World and New World: home grown or translocated? AB - Humans inhabiting the Old World and New World share a wide variety of pathogens. Processes that result in the disjunct biogeographic distribution of pathogens with common vertebrate reservoirs or vectors are more difficult to unravel than those influencing the distribution of infections spread only through human-to human transmission. The origins of species and complexes of tick-borne bacteria are unclear. The agent of Lyme borreliosis may have speciated in the New World following geographical isolation of ticks harboring ancestral spirochetes; the subsequent spread to Europe of B. burgdorferi sensu stricto may have occurred within historical times. Other tick-borne agents, such as the ehrlichiae causing human granulocytic ehrlichiosis, are genetically very similar in the Old World and New World. As the taxonomic distinctions among these related agents of human and veterinary importance appear increasingly blurred, the processes leading to the current discontinuous geographic distributions will also become the source of continuing speculation. Accumulating data suggest an Old World origin for a group of bacteria that include B. elizabethae, a human pathogen first identified from the New World. The potential public health significance of these newly described organisms is undefined, but of international interest as their vertebrate reservoir has been introduced throughout the world. PMID- 10476549 TI - [Evolutionary ecology and epidemiology of interactions between Anopheles mosquitoes and malaria]. AB - The transmission of malaria is largely determined by two parameters: the biting rate of the mosquito vector and its mortality. In this paper, data on the interactions among these parameters are reviewed to describe possible evolutionary mechanisms underlying the parasite's life cycle. In particular, in contrast to conventional wisdom about medical entomology, the author suggests that malaria parasites are not always expected to minimise the damage they inflict on their mosquito host. Rather, when they have developed into the infectious stage, they can increase their transmission by manipulating the mosquito to bite more frequently; this, however, is associated with a higher risk of being killed by the human host. This example illustrates that parameters determining malaria transmission can only be understood by integrating ecological and evolutionary ideas into more traditional epidemiology. Such an evolutionary view of malaria and mosquitoes will eventually lead to a better understanding of the epidemiology of malaria and may help to predict the effect of malaria control. PMID- 10476550 TI - Can molecular biology contribute usefully to vector control? AB - It is argued that further work in molecular entomology should be driven by the practical problems that vector controllers have, not by what molecular biologists can and would like to do. The usefulness is discussed of molecular approaches to (1) insecticide resistance, (2) identification of sibling species, sporozoites and the origin of infections, vector populations and blood meals, (3) genetic control by sterile males or population replacement. PMID- 10476551 TI - [Fulminant septicemia in acute bacterial endocarditis]. PMID- 10476552 TI - When should you refer in suspected endometriosis? PMID- 10476553 TI - A four-year-old boy with hyperactivity. PMID- 10476554 TI - Managing bleeding in pregnancy. PMID- 10476555 TI - Diagnosing and treating polycystic ovary syndrome. PMID- 10476556 TI - Key developments in obstetrics. PMID- 10476557 TI - How long should a woman take HRT? PMID- 10476558 TI - Diagnosis and treatment of chronic pelvic pain. PMID- 10476559 TI - Head lice--developing a practical approach. PMID- 10476561 TI - Preparing for the new format oral exam. PMID- 10476560 TI - Travelling with heart disease. PMID- 10476562 TI - Cervical screening: making it better. PMID- 10476563 TI - A patient with a complaint against a practice partner. PMID- 10476564 TI - Key developments in women's health. PMID- 10476565 TI - A GP's guide to choosing combined pills. PMID- 10476566 TI - Current fibroid management. PMID- 10476567 TI - Managing breast pain. PMID- 10476568 TI - The diagnosis and treatment of hirsutism. PMID- 10476569 TI - Helping women through the perimenopause. PMID- 10476570 TI - When to treat raised lipids. PMID- 10476571 TI - The patient with polyarthralgia. PMID- 10476572 TI - GPs need open-access spirometry in COPD. PMID- 10476573 TI - A post-MI man who questions his medication. PMID- 10476574 TI - Who benefits from oxygen therapy? PMID- 10476575 TI - Key developments in respiratory disease. PMID- 10476576 TI - Addressing the causes of hyperventilation. PMID- 10476577 TI - Tuberculosis: an eminently curable enemy. PMID- 10476578 TI - Managing allergic alveolitis. PMID- 10476579 TI - Pitfalls in the treatment of asthma. PMID- 10476580 TI - Self-management plans in asthma. PMID- 10476581 TI - Breast disease: refer or a reassure? PMID- 10476582 TI - Preparing for multiracial practice. PMID- 10476583 TI - Tinea pedis. PMID- 10476584 TI - A strategy for success in the MRCGP. PMID- 10476585 TI - Models and remodeling: mechanisms and clinical implications. AB - Ventricular remodeling is a process by which the size, shape and composition of cardiac chambers as well as the thickness and composition of the walls are altered in response to physical loads and/or receptor activation, whether created by loss or overload of cardiac myocytes, or the effects of external hormonal or chemical factors. Involved in this process are hypertrophy, dilation, myocyte loss whether due to necrosis or apoptosis, and myocyte hyperplasia. The present review discusses the dynamic and plastic structure of the heart in its capacity to respond to loading. The mechanisms by which myocyte growth as well as myocyte loss are mediated offer therapeutic opportunities to alter these events. PMID- 10476586 TI - Coronary angioplasty and coronary artery remodeling. AB - Coronary artery remodeling is an active process of structural changes in vascular wall structures, in response to varied physiologic or pathologic stimuli. In the past few years, the advent of a new imaging modality such as intravascular ultrasound has allowed in vivo studies, increasing our understanding of this phenomenon. This article reviews some fundamental concepts of coronary vascular remodeling in atherosclerosis and after angioplasty. PMID- 10476587 TI - Homocysteine and risk for atherothrombotic events. AB - Homocysteine is formed during the metabolism of methionine. Plasma concentration of homocysteine depends on the activity of specific enzymes and/or serum concentration of folate, vitamin B6 and B12. Increased levels of plasma homocysteine are known to be an independent risk factor for atherothrombosis. Genetic and non-genetic causes are involved in the etiology of hyperhomocysteinemia. Several biochemical mechanisms underlie the vascular damage and the risk of thromboembolism associated with hyperhomocysteinemia. The overproduction of oxygen free radicals generated from the oxidation of homocysteine may be a major cause of endothelial injury and of the alterations in clotting and vascular function. New strategies have been suggested for the treatment of hyperhomocysteinemia, but more epidemiological data and clinical trials are needed. PMID- 10476588 TI - Use of N-acetylcysteine in the management of coronary artery diseases. AB - Thrombolysis after acute myocardial infarction may lead to a number of adverse effects (reperfusion injury) such as myocardial stunning, arrhythmias and even myocardial damage and extension of the infarct size. Some recent clinical studies have demonstrated that the intravenous infusion of N-acetylcysteine during thrombolysis was associated with a decrease in infarct size and better preservation of left ventricular function, probably due to antioxidant and free radical scavenger properties of N-acetylcysteine. Short- and long-term studies indicated that also in patients with unstable angina pectoris and threat of infarct, the intravenous or oral administration of N-acetylcysteine in association with nitroglycerin is highly effective in decreasing the risk of worsening, mainly by preventing the occurrence of acute myocardial infarction. PMID- 10476590 TI - Dobutamine-induced T wave positivization after uncomplicated myocardial infarction: a marker of myocardial viability and higher cardiac risk. AB - BACKGROUND: There is evidence that after uncomplicated acute myocardial infarction, T wave positivization during stress testing may unveil myocardial viability. We evaluated in a prospective study the clinical value of T wave positivization during dobutamine stress echocardiography in patients with recent, first uncomplicated acute myocardial infarction. METHODS: Two hundred twenty-nine patients, who underwent dobutamine stress echocardiography within 10 days of uncomplicated acute myocardial infarction, were selected for exhibiting negative T waves in the infarct area. A mean follow-up of 2.1 +/- 1 years (up to 6 years) was obtained. RESULTS: T wave positivization appeared during dobutamine test in 76 (33%) patients. The agreement of T wave positivization for myocardial viability was 65% (95% confidence interval 59-71). Compared to myocardial viability during dobutamine stress echocardiography, the combination with T wave positivization was more sensitive (55 vs 24%, 95% confidence interval 46-64 vs 17 33) for predicting cardiac events, albeit less specific. Kaplan-Meier survival curves showed 47 (62%) cardiac events in patients with T wave positivization and 70 (46%) cardiac events in the remaining patients (p < 0.05). Soft (n = 91) prevailed over hard (n = 26) cardiac events. CONCLUSIONS: T wave positivization during dobutamine stress echocardiography after uncomplicated acute myocardial infarction identifies patients at higher cardiac risk, and is more sensitive, albeit less specific, for cardiac events than viability alone. T wave positivization is helpful in the case of inconclusive stress echocardiography. The pathophysiology of T wave positivization and its relative value among other variables warrant further analysis. PMID- 10476589 TI - Usefulness of platelet glycoprotein IIb/IIIa inhibitors in coronary stenting for reconstruction of complex lesions: procedural and 30 day outcome. AB - BACKGROUND: Intracoronary stent implantation during percutaneous transluminal coronary angioplasty (PTCA) has shown favorable results, reducing acute complications associated with PTCA, such as coronary artery dissection and abrupt or threatened vessel closure. However, treatment of lesions with a complex morphology and diffuse disease, requiring long or multiple coronary stents, is still associated with a poorer outcome. We investigated the hypothesis that abciximab might lead to a different outcome in patients with complex coronary lesions, which require long or multiple stent implantation. METHODS: One hundred and six patients were randomized to receive either a combination of abciximab (bolus and 12 hour infusion) and weight-adjusted low-dose heparin or weight adjusted heparin alone and followed up to 30 days. RESULTS: The procedural success rate was 100% in both groups of patients. In the control group a composite rate of major adverse events such as any death irrespective of cause, Q wave or non-Q wave myocardial infarction, acute or subacute stent thrombosis and urgent revascularization of 15.3% was shown at 30-day follow-up. The use of abciximab reduced the composite adverse event rate to 3.7% (76% absolute reduction, p < 0.05). CONCLUSIONS: The use of abciximab during high risk stenting is safe and reduces the risk of cardiac events at 30-day follow-up as compared to standard treatment with heparin. A longer follow-up period is warranted to confirm the beneficial effects observed at 30 days with abciximab in this setting. PMID- 10476591 TI - Magnetic resonance contrast enhancement with gadolinium-DTPA in patients with angina and angiographically normal coronary arteries: effect of chronic beta blockade. AB - BACKGROUND: The syndrome of angina and normal coronary arteries (syndrome X) comprises a heterogeneous group of patients with typical chest pain, a positive exercise test, angiographically smooth coronary arteries and no evidence of spasm. Magnetic resonance imaging (MRI) has been used to detect areas of myocardial ischemia and/or recent necrosis both in animal and human studies. Most of these studies have been conducted after intravenous administration of the paramagnetic contrast medium gadolinium-DTPA (Gd-DTPA), that is considered a sensitive marker of extracellular, probably ischemic in origin, edema. On the basis of these data, we used MRI to evaluate the possibility of myocardial Gd DTPA deposition at rest in patients with syndrome X, and to assess the effects of oral treatment with atenolol. METHODS: We have studied 24 patients with syndrome X, 10 patients with coronary artery disease and 10 age-matched control subjects. The protocol was similar in all study subjects. Exercise testing and MRI were undertaken off therapy after coronary arteriography. Following MRI, patients underwent a 10 day treatment period with atenolol and repeated exercise stress test and MRI while on therapy. RESULTS: In all patients with syndrome X and coronary artery disease were observed effort diagnostic ST-segment changes that were associated with angina in 9 (37%) and 7 (70%) patients, respectively. Of 24 patients with syndrome X, 16 (66.6%) showed areas of myocardial enhancement after Gd-DTPA in comparison to the precontrast imaging. In 4 out of 10 patients with coronary artery disease (40%), Gd-DTPA accumulation was documented. Finally, focal Gd-DTPA myocardial enhancement was not observed in any normal control subject. After beta-blockade, 22 (92%) patients with syndrome X and 2 (20%) with coronary artery disease did not show any ischemic ST-segment changes on effort; 14 syndrome X patients (88%) and 2 coronary artery disease patients (50%) showed complete disappearance of the previously Gd-DTPA enhanced areas on MRI. CONCLUSIONS: Patients with syndrome X often exhibit regional accumulation of Gd DTPA on MRI. The agent is believed to trace interstitial water accumulation as occurs during ischemia and its accumulation is reduced or abolished by treatment with atenolol, probably by different mechanisms. It is likely that an overactivation of the sympathetic outflow to the cardiovascular system can induce most of the abnormalities observed in syndrome X patients. In this context, beta blockers probably represent the mainstay of the medical treatment of this condition. PMID- 10476592 TI - A simple assay for platelet-mediated hemostasis in flowing whole blood (PFA-100): reproducibility and effects of sex and age. AB - BACKGROUND: A simple assay for platelet-mediated hemostasis in flowing blood (PFA 100) has become available. Whole blood is aspirated through the central opening of a membrane coated with platelet agonists; the time required for a platelet thrombus to occlude the opening is defined as closure time; the shorter the closure time the greater the platelet aggregability. There are limited data on the normal range of values for this test, and the effects of sex and age are not known. The aim of this study was to determine the effects of sex and age on closure time in normal volunteers and to assess the reproducibility of the test in our laboratory. METHODS: Closure time using collagen/adenosine-5'-diphosphate was measured in 62 apparently healthy individuals 35 to 75 years of age (11 men and 17 women < 55 years and 22 men and 12 women > 55 years). RESULTS: Closure time was 96.6 +/- 24 s in men and 93.1 +/- 16 s in women (p = 0.20). In the entire group, closure time did not significantly correlate with age (r = -0.17, p = 0.18). However, men < 55 years tended to have a longer closure time than men > 55 years (109.7 +/- 23 vs 90.0 +/- 22 s, p = 0.08), whereas in women closure time was similar in those younger or older than 55 years (93.1 +/- 16 vs 93.3 +/- 18 s, respectively). In 20 samples tested in duplicate, the mean closure time was 80.9 +/- 10 s on the first determination and 81.5 +/- 12 s on the second (r = 0.89, p < 0.001). There was no significant correlation between closure time and hematocrit, platelet number, mean platelet volume, or leukocyte count. CONCLUSIONS: The platelet function analyzer PFA-100 showed a good reproducibility in apparently healthy subjects. No significant difference in closure time was found between men and women, nor between subjects younger or older than 55 years, although a tendency towards shorter values was found in older compared with younger men. PMID- 10476593 TI - [The effects of verapamil on the postischemic changes in the coronary microcirculation: the role of nitric oxide]. AB - BACKGROUND: The aim of this study was to evaluate, in the model of isolated working rat heart, the effects of verapamil on postischemic changes in cardiac mechanical function and microvascular coronary permeability, and the possible role of nitric oxide. METHODS: We used 72 male Wistar rats, weighing 250-300 g, divided into six groups: Group A, hearts perfused with modified Krebs-Henseleit solution (KH); Group B, hearts perfused with KH + verapamil 0.25 microM; Group C, hearts perfused with KH + verapamil 0.5 microM; Group D, hearts perfused with KH + verapamil 1 microM; Group E, hearts perfused with KH + NG-nitro-L-arginine methyl ester (L-NAME) 30 microM; Group F, hearts perfused with KH + verapamil 0.25 microM + L-NAME 30 microM. Hemodynamic parameters, necrosis enzyme release and fluoroscein isothiocyanate-albumin extravasation were evaluated. RESULTS: We observed a clear preservation of cardiac mechanical function and microvascular function in Group B (low dose verapamil) compared to groups A (control), C and D (high dose verapamil); the inhibition of nitric oxide-synthase in the presence of verapamil, obtained in Group F, elicited a loss of myocardial protective effects of verapamil. CONCLUSIONS: Our data suggest that low dose verapamil is effective on postischemic damage reduction and most probably nitric oxide plays a determinant role in this effect. PMID- 10476594 TI - Thrombosis of a mitral valve prosthesis resulting from Staphylococcus epidermidis endocarditis. AB - A 70-year-old man with a Duromedics mitral valve prosthesis had two episodes of infective endocarditis caused by enterococcus (1994 and 1996). Colonoscopy revealed five polyps. Surgical resection was performed and 2 days later the patient had dyspnea and fever. Because of a suspected valve thrombosis, intravenous heparin was given which resulted in hematic effusion in the Douglas' cul-de-sac. Intravenous heparin was withdrawn but the patient continued to have a worsening dyspnea, hyperthermia and hypotension. The patient was transferred to our Institution in cardiogenic shock. Acute thrombosis of the valve was diagnosed by echocardiography, and the patient died before transesophageal Doppler echocardiography was performed. Post mortem examination revealed mitral valve infective thrombosis. In patients with valvular prostheses, endocarditis is an added thromboembolic risk. PMID- 10476595 TI - The coronary circulation: quo vadis? AB - This review examines past and present observations concerning the structure and function of the coronary circulation in health and disease. These observations are considered in the context of how we might proceed to increase understanding of this circulation in the future. The coronary microcirculation with its intimate relationship of capillaries and myocytes and the coronary venous circulation are identified as parts of the coronary circulation in need of further study. Observations using a laser Doppler velocimeter placed within the beating myocardium are presented. Such devices measure the velocity of red cells continuously (red cell flux) and can demonstrate that tissue hematocrit and hence oxygen delivery can be regulated independently of total epicardial arterial volume flow. Implications for the understanding of the pathophysiology of coronary artery disease are presented. As considerations are given to molecular genetic techniques to revascularize the ischemic myocardium we will require a more complete knowledge of coronary circulatory dynamics, myocardial support tissues' responses, and cardiac myocyte interactions to design appropriate interventions. The clinical trial is an appropriate clinical tool to measure effectiveness but a blunt instrument to determine pathophysiology. The purpose of the review is to suggest that advances in measurement of end points are required to permit the right question to be posed. PMID- 10476596 TI - Abdominal aortic aneurysms: current management. AB - Optimal management of abdominal aortic aneurysm (AAA) remains a challenging surgical problem. Over the last decade surgical and anesthetic improvements have provided perioperative mortality in the 2% range, when elective AAA repair was performed in single Institutions with large vascular experience. However, community- or national-based mortality rates for elective AAA surgery may be as high as 11% or more. Mortality rates associated with ruptured aneurysms remain as high as 90%. AAA prophylactic resection should be indicated when the risk of rupture exceeds the surgical risk. Although the risk of rupture correlates strongly with the diameter of the AAA, there is evidence that other factors can increase the rupture risk: hypertension, chronic pulmonary disease, aneurysm morphology, etc. Establishing a single threshold diameter for AAA repair appears naive. Moreover, AAA primarily affects older patients with other comorbidities that shorten life expectancy and increase perioperative risks: coronary artery disease, renal and pulmonary insufficiency, peripheral artery disease, etc. So that, proper management of individual AAA is based on balancing the perioperative risk, the risk of rupture, and life expectancy. In the subgroup of young healthy patients with additional risk factors for AAA rupture, elective repair at a smaller size (4 to 5.5 cm) may be beneficial if low surgical risk can be assured. In the last decade endovascular repair for AAA treatment has emerged. These less invasive endovascular techniques for AAA repair offer some advantages in terms of reduced patient stress, analgesic requirement, respiratory dysfunction, blood loss, need for intensive care and reduced hospitalization with an early technical success similar to that of open surgical treatment. However, there are no prospective, randomized studies evaluating endovascular treatment of AAA. Moreover, long-term results on the durability of these new techniques are needed to assess endovascular repair as an alternative treatment to prevent the risk of AAA rupture. PMID- 10476597 TI - Abnormal transport of inorganic phosphate in left ventricular mitochondria from spontaneously hypertensive rats. AB - INTRODUCTION: The aim of this study was to investigate the kinetic properties of inorganic phosphate (Pi) translocator in intact mitochondria isolated from the hypertrophied left ventricular tissue of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) at the ages of 5 and 24 weeks, before and after the development of hypertension. METHODS: The dependence of the Pi uptake rate on substrate concentration was measured in both absence and presence of mersalyl by spectroscopic techniques. RESULTS: Saturation characteristics were found (Km 250.0 +/- 25.0 and 15.0 +/- 1.5 microM for 5- and 24-week-old SHR, and 300.0 +/- 30.0 and 40.0 +/- 4.5 microM for WKY rat mitochondria, respectively, p < 0.05; Vmax 1.2 +/- 0.16 and 0.1 +/- 0.01 delta A/min x mg mitochondrial proteins for 5- and 24-week-old SHR, and 4.1 +/- 0.39 and 1.4 +/- 0.12 delta A/min x mg mitochondrial proteins for 5- and 24-week-old WKY rats, respectively, p < 0.05). When Pi carrier activity was measured using concentrations which are assumed to be in the cytosol under physiological conditions, Pi carrier velocity was 1.1 and 0.1 in SHR and 4.6 and 1.4 delta A/min x mg mitochondrial proteins in WKY, at 5 and 24 weeks, respectively. CONCLUSIONS: The significant decrease in the activity of the Pi carrier could imply that pressure overload is critical in SHR. Nevertheless, as decreased activity was found in SHR also at an early age when animals do not show stable increased blood pressure levels, we suggest that other factors might contribute to the abnormalities of Pi transport in mitochondria. An altered gene expression possibly related to a primary defect in this strain or, alternatively, to an abnormal regulation of protein synthesis might be proposed as additional factors affecting Pi carrier activity. The results of this study, together with previous data of the literature showing abnormalities in energy production mechanisms, allow us to hypothesize a profound rearrangement of energy metabolism at the mitochondrial level in this model of left ventricular hypertrophy and hypertension. PMID- 10476598 TI - Diagnostic value of exercise electrocardiography for predicting a positive scintigraphic test in patients with complete right bundle branch block. AB - BACKGROUND: The diagnostic value of exercise electrocardiography (ECG) in patients with complete right bundle block (cRBBB) remains controversial. The aim of this study was to investigate the diagnostic accuracy of exercise ECG for predicting ischemia in the presence of cRBBB. METHODS: From a series of 1300 consecutive patients attending for exercise ECG and 99mTc-tetrofosmin single photon emission computed tomography (SPECT), we identified 38 male patients with cRBBB and 38 matched controls with normal intraventricular conduction. Patients with left ventricular hypertrophy or previous revascularization were excluded. Exercise ECG (modified Bruce protocol) was considered positive at > or = 1 mm ST horizontal or downsloping depression in > or = 2 adjacent leads. SPECT imaging at rest and post-exercise was performed at least 48 hours apart. RESULTS: The odds ratio for ischemia comparing patients with positive and negative exercise ECGs was 11.0 (95% confidence interval 2.49-48.64, p = 0.002) in controls and 2.49 (95% confidence interval 0.64-9.08, p = 0.32) in cRBBB. The prior probability of ischemia was 0.37 in controls and 0.58 with cRBBB. The posterior probability after a positive test was 0.65 in controls and 0.68 in cRBBB. Thus, the utility of the test (posterior probability minus prior probability) was greater in controls (+0.28) than in cRBBB (+0.1). This difference was most pronounced in the anterior leads V1-V4 (controls +0.63 vs cRBBB +0.01) but less significant in the lateral leads V5-V6 (controls +0.26 vs cRBBB +0.21). Similar analysis also indicated reduced diagnostic value of negative exercise ECGs in cRBBB patients. CONCLUSIONS: The diagnostic value of exercise ECG is reduced in patients with cRBBB, although ST-segment changes in the lateral ECG territory provide superior predictive accuracy to those in the anterior leads. The use of SPECT as a first line investigation may be justified in patients with cRBBB. PMID- 10476599 TI - [The diagnostic and therapeutic procedures in pulmonary embolism: a survey in the Veneto Region]. AB - BACKGROUND: Pulmonary embolism is one of the most frequent cardiopulmonary diseases, but it is often under- or misdiagnosed. In order to address this issue and to identify flow charts that are commonly used in pulmonary embolism diagnosis and treatment, 191 clinical wards of internal medicine, cardiology, geriatrics, pneumology and intensive care units, located in the Veneto Region, were surveyed. METHODS: An anonymous questionnaire was mailed to each ward in order to collect clinical diagnostic information on all pulmonary embolisms which occurred during 1993. Among the returned questionnaires, 114 (59.6%) had usable information for the analysis. RESULTS: The vast majority of participating centers reported in 1993 less than 10 pulmonary embolism events. No significant differences were observed between internal medicine, geriatrics, pneumology wards and intensive care units. The reported events, however, were slightly higher in the divisions of cardiology with an annual average of 12 events per center. First level diagnostic procedures, such as ECG, chest X-ray and arterial blood gas analysis were chosen and performed in all patients. Interestingly, Doppler echocardiography, which is often not included in official guidelines for pulmonary embolism diagnosis, was performed in 56% of the participating centers. On the contrary, ventilation-perfusion lung scanning, which is considered highly predictive in many diagnostic algorithms, was underutilized (35% perfusion scan, 20% ventilation scan). This underuse was probably due to technical and organizational difficulties. Pulmonary angiography, the most accurate procedure for the diagnosis of pulmonary embolism, was performed in 28% of the patients. During the acute phase, intravenous heparin was commonly used; 91% of patients received the infusion continuously, 4% intermittently. Thrombolysis was performed in 25% of the patients. The preferred drugs were recombinant tissue-plasminogen activator (67%), followed by urokinase (20%) and streptokinase (13%). To start thrombolytic therapy, 20% of the interviewed clinicians considered sufficient the evidence of clinical manifestations of pulmonary embolism confirmed by echocardiographic data. At discharge, prescription of oral anticoagulant drugs was common (78%) for at least 6 months (47%). Standardized procedures for the diagnosis and treatment of pulmonary embolism were already implemented in 13% of the participating centers. CONCLUSIONS: These data suggest a common effort to define unanimous conventional protocols in the management of pulmonary embolism. It should be underlined, however, that a particular attention to the clinical manifestations and a productive collaboration among clinicians with different expertise are required to improve the diagnosis and treatment of pulmonary embolism. PMID- 10476600 TI - Pulmonary embolism: opinions and facts. PMID- 10476601 TI - Automatic activity of a decremental long atrioventricular accessory pathway: an unusual feature. AB - The electrophysiological properties of decremental atrioventricular and atriofascicular pathways are not completely understood. We report the case of a patient with fast reentrant tachycardia due to a decremental long atrioventricular pathway, who showed a slow automatic tachycardia arising from the same pathway that was successfully eliminated by radiofrequency catheter ablation. PMID- 10476602 TI - [Recommendations for cardiovascular assessment and risk reduction in diabetes- 1999. Societa Italiana di Diabetologia, the Diabetes and Atherosclerosis Study Group, Associazione Medici Diabetologi]. PMID- 10476603 TI - Experiments carried out over the Web. PMID- 10476604 TI - Illusory inferences: a novel class of erroneous deductions. AB - The mental model theory postulates that reasoners build models of the situations described in premises, and that these models normally make explicit only what is true. The theory has an unexpected consequence: it predicts the occurrence of inferences that are compelling but invalid. They should arise from reasoners failing to take into account what is false. Three experiments corroborated the systematic occurrence of these illusory inferences, and eliminated a number of alternative explanations for them. Their results illuminate the controversy among various current theories of reasoning. PMID- 10476605 TI - The relationship between language-processing and visual-processing deficits in developmental dyslexia. AB - Some research on developmental dyslexia focuses on linguistic abnormalities such as poor reading of nonwords or poor reading of exception words. Other research focuses on visual abnormalities such as poor performance on psychophysical tasks believed to assess the functioning of the magnocellular and parvocellular layers of the lateral geniculate nucleus (LGN). Little is known about what the relationships are between these two types of abnormalities. We measured nonword reading, exception word reading, and performance with Ternus apparent movement displays (the perception of which is believed to depend upon the magnocellular and parvocellular pathways) in dyslexic children and children without reading difficulties. Our results indicate that performance on the Ternus task is related to nonword reading ability but not to exception word reading ability. We offer two alternative interpretations of these findings. According to the first of these, nonword reading requires a serial left-to-right allocation of covert attention across the letter string being read and the neural systems involved in this attentional process also play a part in responses to the Ternus display. According to the second, poor nonword reading and abnormal Ternus performance are not directly related: perinatal/neurodevelopmental insult has affected the LGN (influencing Ternus performance) and the adjacent medial geniculate nucleus (MGN; affecting phonological ability) and the MGN abnormalities may be more functionally related to poor nonword reading. PMID- 10476606 TI - Synchronous change and perception of object unity: evidence from adults and infants. AB - Adults and infants display a robust ability to perceive the unity of a center occluded object when the visible ends of the object undergo common motion (e.g. Kellman, P.J., Spelke, E.S., 1983. Perception of partly occluded objects in infancy. Cognitive Psychology 15, 483-524). Ecologically oriented accounts of this ability focus on the primary of motion in the perception of segregated objects, but Gestalt theory suggests a broader possibility: observers may perceive object unity by detecting patterns of synchronous change, of which common motion is a special case. We investigated this possibility with observations of adults and 4-month-old infants. Participants viewed a center occluded object whose visible surfaces were either misaligned or aligned, stationary or moving, and unchanging or synchronously changing in color or brightness in various temporal patterns (e.g. flashing). Both alignment and common motion contributed to adults' perception of object unity, but synchronous color changes did not. For infants, motion was an important determinant of object unity, but other synchronous changes and edge alignment were not. When a stationary object with aligned edges underwent synchronous changes in color or brightness, infants showed high levels of attention to the object, but their perception of its unity appeared to be indeterminate. An inherent preference for fast over slow flash rates, and a novelty preference elicited by a change in rate, both indicated that infants detected the synchronous changes, although they failed to use them as information for object unity. These findings favor ecologically oriented accounts of object perception in which surface motion plays a privileged role. PMID- 10476607 TI - Annual report of the National Influenza Surveillance Scheme, 1998. AB - The National Influenza Surveillance Scheme includes data from sentinel general practice consultations for influenza-like illness, laboratory reports of influenza and absenteeism rates from a national employer. The 1998 season was dominated by an increase in influenza A in all States and Territories and low influenza B activity. All influenza A isolates were characterised as influenza A (H3N2). Peak activity in 1998 was recorded in July and August. Data are coordinated, analysed and disseminated at a national level and published in Communicable Diseases Intelligence during the influenza season. PMID- 10476608 TI - Annual report of the Australian Gonococcal Surveillance Programme, 1998. AB - The Australian Gonococcal Surveillance Programme examined 3,583 isolates of Neisseria gonorrhoeae in 1998. Again in 1998 the rates and sites of infection and antibiotic susceptibility patterns varied considerably between regions, reflecting considerable differences between rural and urban gonorrhoea in Australia. Resistance to the penicillin and quinolone groups of antibiotics was highest in urban centres, but penicillins remained suitable for use in many parts of rural Australia. Quinolone-resistant gonococci continued to be concentrated in New South Wales (NSW) where sustained domestic transmission of these strains was maintained but at a lower rate than in 1997. Endemic transmission of Quinolone resistant gonococci in homosexually active men was found for the first time. Quinolone-resistant gonococci in other centres continued to be isolated mostly from overseas travellers and at a lower frequency. All isolates remained sensitive to spectinomycin and ceftriaxone. Strains showing high level tetracycline resistance increased by 300% in NSW and were acquired predominantly through local contact. A significant increase in the number of isolates was recorded in NSW and Victoria in 1998, this increase being mainly attributable to an increase in gonorrhoea in homosexually active males. Strains examined in South Australia (SA), NSW and Victoria were predominantly from male patients and rectal and pharyngeal isolates were common. In other centres the male to female ratio was lower, and most isolates were from the genital tract in rates similar to those occurring in 1997. PMID- 10476609 TI - Rotavirus diversity: what surveillance will tell us. PMID- 10476610 TI - Enterovirus 71 outbreak in Western Australia associated with acute flaccid paralysis. Preliminary report. PMID- 10476612 TI - Meeting summary for NASSHP, July 17-19, 1998. North American Society for the Study of Hypertension in Pregnancy. PMID- 10476611 TI - Communicable diseases surveillance. PMID- 10476613 TI - Nitric oxide synthase activity and Doppler parameters in the fetoplacental and uteroplacental circulation in preeclampsia. AB - OBJECTIVE: We investigated the hypothesis that changes in blood flow in the uteroplacental and fetoplacental circulation in preeclampsia are associated with an abnormality of placental or uterine placental bed nitric oxide (NO) synthesis. METHODS: We measured pulsatility indices on Doppler waveform analysis from uterine and umbilical arteries in 20 patients with preeclampsia and 14 healthy pregnant controls before elective cesarean section. During cesarean section, biopsies from the uterine placental bed and the placenta were taken and the nitric oxide synthase (NOS) activity was measured by the [3H] L-arginine-[3H] L citrulline conversion assay in these samples. RESULTS: The NOS activity was significantly lower in the uterine placental bed in comparison to the placental tissue (p < 0.01). Placental NOS activity was similar between patients with preeclampsia and healthy controls and in the groups with either a pathological or a normal Doppler flow in the umbilical artery. In the uterine placental bed however, NOS activity from patients with preeclampsia was significantly lower (p < 0.01), whereas the blood flow resistance in the uterine arteries was elevated (p < 0.01) in comparison to healthy controls. CONCLUSIONS: Our data show that pathological Doppler waveforms in the uterine arteries of patients with preeclampsia are paralleled by diminished NOS activity in the uterine placental bed. Therefore, the compromised NO production in the uterine placental bed may play an important role in the impaired uteroplacental blood flow and potentially in some pathological features of preeclampsia such as intervillous thrombosis formation and fetal growth retardation. PMID- 10476614 TI - Renal handling of endogenous human chorionic gonadotrophin in preeclampsia. AB - OBJECTIVE: To test the hypothesis that the elevated maternal serum concentration of human chorionic gonadotrophin (hCG) in preeclampsia is due to altered renal handling of the hormone. SETTING: Department of Obstetrics and Gynecology, Al Ain Hospital, United Arab Emirates, a tertiary center affiliated with the Faculty of Medicine and Health Sciences, UAE University. METHODS: The renal clearances and handling of endogenous creatinine and human chorionic gonadotrophin were compared in 14 normotensive volunteer and 14 preeclamptic mothers who received oral hydration at 34-37 weeks' gestation. The hCG content in the placentas was estimated immunohistochemically after delivery. RESULTS: Maternal serum concentration of hCG (p = 0.0057), the placental hCG immunopositive cell (p < 0.0001), and syncytial knot counts (p < 0.0001) were significantly higher in preeclamptic mothers. The renal clearances of endogenous creatinine and hCG and fractional hCG clearance were not significantly different in both groups. Significantly increased amounts of hCG were filtered (p = 0.007) and excreted (p = 0.007) by preeclamptic mothers. Only a small but fixed proportion of the filtered load of hCG is excreted in both groups and there was a positive correlation (r = 0.5, p = 0.005) between filtered and excreted loads of hCG. CONCLUSION: The results indicate increased placental content of hCG in preeclampsia. The resultant increased maternal serum concentration is probably sustained by the mechanism of renal handling of the hormone. PMID- 10476615 TI - Serum and placental lipid peroxides in chronic hypertension during pregnancy with and without superimposed preeclampsia. AB - OBJECTIVE: To evaluate the circulating levels of lipid peroxides and vitamin E, and the placental levels of lipid peroxides in chronic hypertensive pregnant women, with and without superimposed preeclampsia, as compared to controls and women with primary preeclampsia. METHODS: Lipid peroxides were measured in serum and placenta by the thiobarbituric acid method and high-pressure liquid chromatography (HPLC), and vitamin E by HPLC. Patients were 36 healthy pregnant women, 34 previously nonhypertensive women diagnosed with preeclampsia, 20 women with uncomplicated chronic hypertension, and 11 women with chronic hypertension complicated by superimposed preeclampsia. RESULTS: Lipid peroxides in serum and placental tissue were significantly increased, and vitamin E levels in serum were significantly decreased in women with primary preeclampsia and superimposed preeclampsia, as compared to controls. The group of uncomplicated chronic hypertension presented with similar values of lipid peroxides and vitamin E to controls. CONCLUSIONS: Our results support the clinical assumption that chronic hypertension aggravated by the development of proteinuria represents a superimposed condition associated with placental disease. The data further support the concept that increased lipid peroxides are not merely associated with the presence of hypertension in pregnancy, but they are implicated in the pathophysiology of preeclampsia. PMID- 10476616 TI - Plasma glutathione S-transferase Pi 1-1 measurements in the study of hemolysis in hypertensive disorders of pregnancy. AB - OBJECTIVE: To investigate the value of plasma glutathione S-transferase Pi1 1(GSTP1-1) measurements in the assessment of hemolysis in hypertensive disorders of pregnancy. METHODS: Plasma GSTP1-1 and haptoglobin levels and serum lactate dehydrogenase (LDH) activity were measured in 81 healthy nonpregnant female blood donors between 20 and 40 years of age, 41 women during uncomplicated normotensive pregnancy, 35 women with pregnancy-induced hypertension, 67 women with preeclampsia, and 34 women with the HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome. Levels in hypertensive pregnancies were compared to levels in normotensive pregnancy, and levels in normotensive pregnancy were compared to levels in blood donors by the Mann-Whitney U-test. RESULTS: Median GSTP1-1 and LDH levels were significantly increased (p < 0.01) and haptoglobin significantly decreased (p < 0.01) in preeclampsia and the HELLP syndrome as compared to normotensive pregnancy. Both GSTP1-1 and LDH levels were significantly higher in normotensive pregnant women as compared to nonpregnant women (p < 0.0001). The percentage of preeclamptic patients (26.9%) or patients with the HELLP syndrome (73.5%) with elevated GSTP1-1 levels was lower than those with elevated LDH (38.8% and 100%, respectively) or decreased haptoglobin levels (41.8% and 97%, respectively). CONCLUSIONS: We conclude that plasma GSTP1-1 levels may provide useful information on hemolysis in hypertensive disorders of pregnancy in addition to serum LDH activity and plasma haptoglobin levels and that the degree of hemolysis in hypertensive disorders of pregnancy, especially in the HELLP syndrome, is probably less prominent than generally assumed. PMID- 10476617 TI - Trial of prophylactic administration of TXA2 synthetase inhibitor, ozagrel hydrochloride, for preeclampsia. AB - OBJECTIVE: In an attempt to investigate the prophylactic effect of a thromboxane A2 (TXA2) synthetase inhibitor on pregnant women with a high risk of preeclampsia, the following clinical study was undertaken. METHODS: Forty pregnant women were randomly allocated to control or treatment groups. Ozagrel Hydrochloride (400 mg/day, orally) and placebo were started at 20 weeks of gestation and continued until delivery. RESULTS: Seventeen of 20 (85%) women in the control group developed preeclampsia, whereas 9 of 20 (45%) in the treatment group developed preeclampsia. Ozagrel Hydrochloride significantly (p < 0.01) reduced the occurrence of preeclampsia, and the incidence of both hypertension (p < 0.05) and proteinuria (p < 0.01) was significantly less in the treatment group compared with the control group. One month after administration, the mean plasma concentration of TXB2, a metabolite of TXA2, was significantly decreased (p < 0.01) to 62.4 +/- 13.6%, whereas that of 6-keto prostaglandin F1 alpha, a metabolite of PGI2, was significantly increased (p < 0.01) to 206.7 +/- 52.8%. There were no maternal or fetal side effects observed. CONCLUSIONS: It seems likely that Ozagrel Hydrochloride could be used for the prevention of preeclampsia in high-risk pregnant women. PMID- 10476618 TI - Calcium and low-dose aspirin prophylaxis in women at high risk of pregnancy induced hypertension. AB - OBJECTIVE: The objectives of the study were to confirm the validity of using oscillometric measurement of MAP in the left lateral position to identify those at high risk for developing pregnancy-induced hypertension (PIH), and to assess and compare the efficacy of prophylaxis with low-dose aspirin or calcium supplementation in high-risk patients. STUDY DESIGN: A prospective study in pregnancy; 500 normotensive, primigravid Chinese women were recruited in the second trimester of pregnancy on the basis of 80 mm Hg > or = MAP < 106 mm Hg in the antenatal clinic. They were then screened by Dinamap in a research setting, measuring MAP in the left lateral position after rest and using a cutoff value of 60 mm Hg for inclusion in the randomized study. Randomization was divided into three groups: control, low-dose aspirin, and calcium supplementation. After delivery, patients were classified as either having remained normotensive or having developed PIH, with or without proteinuria. RESULTS: The incidence of both proteinuric and nonproteinuric PIH was significantly lower in patients screened out as low risk than in those selected as high risk using a critical value of 60 mm Hg for left lateral MAP (p < 0.05). The incidence of proteinuric PIH was significantly lower in patients given low-dose aspirin than in the control group (p < 0.05). However, the confidence intervals for the effect were wide, comparable with aspirin having no effect or leading to a 16-fold reduction in the risk of preeclampsia. For those given calcium supplementation, the reduction was not significant. There was no significant difference in the incidence of nonproteinuric PIH between the control group and the two groups receiving prophylaxis. CONCLUSION: Oscillometric measurement of second-trimester left lateral MAP is a valid predictor of proteinuric PIH. Low-dose aspirin may offer a degree of protection from proteinuric PIH in these high-risk women. Calcium supplementation was not shown to significantly reduce the incidence of PIH. PMID- 10476619 TI - Blood pressure and vasoactive hormones in mild preeclampsia and normal pregnancy. AB - OBJECTIVE: Changes in vasoactive hormones are reported to play an important role in the pathogenesis of preeclampsia linking placental hypoperfusion with hypertension, systemic disease, and proteinuria. We, therefore, studied diurnal patterns of vasoactive hormones in mild preeclampsia. METHODS: Venous blood samples were drawn every 2 h over 25 h for measurements of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), norepinephrine, renin activity, and aldosterone and two urine collections (12 h each) for stable prostaglandin metabolite measurements. The patients were nine women with mild preeclampsia and, for comparison, nine control women matched for gestation and parity. RESULTS: Women with mild preeclampsia had higher norepinephrine levels throughout 25 h, and lower systemic prostacyclin production as measured by the urinary 2,3-dinor-6 keto PGF1 alpha excretion during the daytime. Plasma endothelin and ANP and BNP concentrations tended to be higher throughout 25 h in preeclampsia, but differences between the two groups did not reach levels of statistical significance. Plasma renin activity and aldosterone did not differ between the groups. Whereas control women exhibited a diurnal variation in plasma norepinephrine, ANP, BNP, and aldosterone, and in both urinary prostacyclin and thromboxane A2 metabolites, this was less distinct or absent in patients with mild preeclampsia. CONCLUSIONS: We conclude that mild preeclampsia is associated with elevated plasma norepinephrine levels, lower systemic daytime production of prostacyclin, and blunting of the normal diurnal variation for a number of indices including plasma levels of BNP, ANP, norepinephrine, and aldosterone, and urinary prostacyclin metabolites. PMID- 10476620 TI - Using baseline data to improve care of the patient with type 2 diabetes mellitus. AB - Diabetes mellitus affects approximately 16 million people in the United States. Despite the publication of multiple guidelines, the delivery of care has not substantially changed. This article describes a program at Advocate Health Care to improve the delivery of care to individuals with Type 2 diabetes mellitus in an ambulatory setting. Using a variety of continuous quality improvement tools, this project enhances partnerships between patients and health care providers. PMID- 10476621 TI - Patient satisfaction with emergency department triage nursing care: a multicenter study. AB - This descriptive, correlational study examines relationships between (1) individual patient and nurse characteristics and (2) patient satisfaction with triage nursing care, patient satisfaction with the triage nurse, and patient's intention to return to a specific Emergency Department. The convenience sample consisted of Urgent/Delayed patients (N = 378) triaged in an urban academic medical center, a public hospital in a small city, and a Catholic hospital in a small city. Analysis of variance revealed significantly higher levels of patient satisfaction at the academic medical center, whereas higher levels of intent to return were reported by subjects from the Catholic-affiliated hospital. Educational preparation of the triage nurse was identified as a significant predictor of both patient satisfaction with triage nursing care and loyalty to a specific. PMID- 10476622 TI - Preventing adverse outcomes: a population focus. AB - This article discusses the concept of risk from an epidemiological viewpoint as well as that of vulnerable populations. Three components of resource availability are pertinent to the concept of risk of a population: social status, social capital, and human capital. These components are illustrated through an example of a project to reduce preventable adverse health outcomes and increase competency in self-care among residents of an inner-city community. Activities and outcomes of the project are discussed. PMID- 10476623 TI - Improving patient care through implementation of nurse-driven restraint protocols. AB - Nationally, much attention has been placed on the indiscriminate application and abuse of restraint usage. This was the impetus for health care institutions across the country to relook at the policy, practices, and procedures regarding restraints. Our health care system made changes to our restraint policy, practice guidelines, and procedures in an effort to assure protection of the patients' health and safety while preserving their dignity, rights, and well-being. The mission was to pursue a restraint-appropriate environment by restraining only those patients who were assessed as being at risk of harming self and to protect the patient or others from injury. Our overall goal was to reduce restraint usage. This article describes the current policies, practice guidelines, and procedures for identifying clinically appropriate and adequately justified situations for restraint usage. The focus is on implementation of nurse-driven restraint protocols to improve patient care. All efforts directed at improvements in restraint usage and management of a patient in restraints has reduced our overall numbers of patients in restraints as well as significantly reduced risk of incidence for patients in restraints. PMID- 10476624 TI - Reengineering the role of a nurse manager in a patient-centered care organization. AB - The Department of Veterans Affairs Medical Center in New York City expanded the role of a nurse manager to a manager and leader of the interdisciplinary team within a structure reorganized to focus on patients. As the literature noted the critical nature of the role of a middle manager, an interdisciplinary team reached the consensus that a registered nurse with progressive clinical and head nurse experience should be the first-line manager in a patient-centered care organization. The process of reengineering the role of a Patient Care Team Coordinator (PCTC), the support systems designed to develop the new leadership role, and the benefits associated with the changes implemented are discussed. The functional statement for the PCTC's position is included, as well as an organizational structure to show how staff are grouped to promote the continuity and coordination of care provided to patients. PMID- 10476625 TI - Analyzing hospital readmissions using statewide discharge databases. AB - Hospital readmissions are an important indicator of the outcomes of care as well as a source of unnecessary health care expenditures. This study focused on development of a uniform algorithm for identification of hospital readmission data. It involved development of a uniform definition of readmissions which could be applied to multiple statewide computer databases. Through this approach, comparable readmit data were generated for use in benchmarking and quality improvement activities. PMID- 10476626 TI - Relationships among educational material readability, client literacy, perceived beneficence, and perceived quality. AB - Illiteracy remains a pervasive problem that compromises quality health care, limits understanding of health information, and potentially leads to poor health outcomes. This study demonstrated that participants' measured reading level was significantly lower than both their stated years of education and readability of an educational pamphlet. A positive relationship was identified between measured reading level and pamphlet helpfulness (client-perceived beneficence). Participants perceived care quality as good to very good. Quality client education requires use of either educational materials appropriate for the measured reading level of clients or alternatives to written material. PMID- 10476627 TI - Tilling the soil: nurturing the seeds of patient and family education. AB - Effective patient and family education has become most essential in today's changing health care environment. The importance of patient education is reflected in the organizational performance standards of the Joint Commission on Accreditation of Healthcare Organizations. To keep abreast of these rapid changes, we decided to evaluate and enhance our existing cardiac education program. This article presents our analysis that effective patient and family education must be a coordinated effort across the cardiac continuum. New teaching tools were established to assist in increasing the patients' knowledge base, and consequently empower patients to make better decisions regarding their care. PMID- 10476628 TI - [The importance of modernization and the role of the scientific society. II. The course of modernization of continuous certification in anesthesia and resuscitation. The mandate of SIAARTI/AAROI. Documentation of the provisional regulation of ACC (awaiting the conclusion of the experimental phase)]. PMID- 10476629 TI - [The role of immunological mechanisms in the development of the late sequelae of nuclear disasters]. AB - Effects, both specific and non-specific, were analysed of ionizing radiation in a comparative evaluation of medical sequels of the Chernobyl disaster and of the Semipalatinsk Range nuclear test. A hypothesis is advanced assuming that development of all major bodily morbid conditions is directly related to the dysfunction of certain complex of organs, the thyroid gland in the first place, the thymus gland known to be a central organ of systemic immunogenesis. A breach in the thymus link triggers harmful cascade reactions of neuroimmunoendocrinological regulation, appeared to be responsible for the pathogenesis of practically all pathologies disclosed. PMID- 10476630 TI - [Changes in the L-serine and L-threonine dehydrogenase activities in the blood serum of those who worked in the cleanup of the aftermath of the accident at the Chernobyl Atomic Electric Power Station who became ill with chronic acalculous cholecystitis]. AB - Activity was studied of blood serum plasmic enzymes L-serine and L-threonine dehydrogenazes (SDG and ThDG) in 92 liquidators of aftermath of the Chernobyl atomic power plant breakdown, presenting with chronic non-calculous cholecystitis during the stage of moderately severe exacerbation with no clinical and laboratory and sonographic signs of affection of the liver. A quarter of the examinees demonstrated an increased activity of the enzymes under study, which fact is regarded by the authors as a preclinical sign of reactive hepatitis. Recommendations are given as to the outpatient registration and prophylactic management and therapy of those persons having taken part in the elimination of the effects of the Chernobyl accident, presenting with biliary pathologies. PMID- 10476631 TI - [The indices of the peripheral blood leukocytes in subjects working within a 30 kilometer area of the Chernobyl Atomic Electric Power Station and in the workers of a chemical plant]. AB - Changes in leukocytes are analyzed together with the pattern of leukogrammes, morphological and cytochemical parameters characterizing leukocytes of the peripheral blood in those persons working within the zone of radiation control and in workers of chemical plants. Criteria of high informative value have been established for identification of those groups of individuals running a risk for development of hematological pathology. PMID- 10476633 TI - [The indices of the reproductive function status of the female population of the Kryvyy Rih basin]. AB - A study was made on the disordered course of pregnancy, complicated labour, and pathological processes in those parturient women being in labour for the first time in their life residing in relatively pure and contaminated regions of Krivoi Rog. There were three age groups of parturient women: those ranging between 18 to 25 years, 26 to 30 years old, and greater than 30 years old. Most commonly seen in the Krivoi Rog industrial region is an increase in chronic intrauterine hypoxia of the fetus together with anemias of pregnancy, with the rise of the above pathologies being recordable both by years (1991-1995) and contaminated region compared to the relatively pure one. PMID- 10476632 TI - [The dynamics of the catecholamine level in the biological fluids of patients with chronic posttraumatic osteomyelitis living in different ecological regions]. AB - Overall, sixty-seven patients living in different ecological regions were examined to study effects of ecological factors on the catecholamine concentration in biological fluids in patients with chronic posttraumatic osteomyelitis (CPO). The data obtained showed that CPO causes abnormalities of catecholamine metabolism, which get even more aggravated under the influence of an adverse ecological factor, an abnormal radiation impact in particular, this being confirmed by analysis of findings in those patients residing in different ecological regions. PMID- 10476634 TI - [Sanatorium-health resort treatment in the correction of the functional disorders in children with cardiovascular pathology who live in a large industrial city]. AB - The incidence rate of functional disorders in the cardiovascular system as well as in the vegetative nervous system was found out to be on the increase in those children with cardial pathology residing in a large industrial centre. Sanatorium health resort treatment was noted to make for improvement of vegetative regulation of the cardiac activity, enhancement of compensatory-and-adaptive potentialities of the organism, which facts warrant a wider use of health resorts in the rehabilitative treatment of children experiencing additional environmental loads of a large industrial centre. PMID- 10476636 TI - [The diagnostic value of different methods for determining Helicobacter pylori]. PMID- 10476635 TI - [The diagnostic criteria of diphtherial myocarditis]. PMID- 10476637 TI - [Estrogen receptors and stomach cancer (a review of the literature and the author's own studies)]. PMID- 10476638 TI - [The characteristics of the course of glomerulonephritis in patients in different age groups]. PMID- 10476639 TI - [The current drug treatment of patients with cerebral venous dystonia]. PMID- 10476640 TI - [The pathophysiology of endogenous intoxication in gestoses]. PMID- 10476641 TI - [Recombinant erythropoietin and early preterm neonatal anemia]. PMID- 10476642 TI - [Low-intensity electromagnetic radiation in medicine: a factor in resonance therapy or a nonspecific adaptogen?]. PMID- 10476643 TI - [The effect of the thrombocytes on the formation of the hemostatic potential in the venous and arterial beds in stable stenocardia]. AB - The coagulative activity was studied of the arterial and venous blood thrombocyte and thrombocyte-free plasma in 16 patients with stable angina. Changes in the general index of thrombophilia permitted the identification of activation of the hemostatic potential in the venous and arterial beds. The results secured suggested the important part that platelets have in formation of the hemostatic potential in both the venous and arterial beds in stable stenocardia. Discussed in the article is the role of the lungs in stabilization of the hemostatic potential in the arterial bed. PMID- 10476644 TI - [The morphometric indices of the myocardium in young hypertension patients with excessive body mass]. AB - Morphometric indices were measured for the myocardium with the aid of echocardiography in 25 young hypertensives, their age ranging between 15 to 30 years. Of these, 4 had normal body mass, 21-above normal. A significant augmentation in the thickness of the left ventricular posterior wall was measurable as was that of the diameter of the left auricular cavity and left ventricular end-diastolic dimension in overweight patients, which fact suggests early formation of hypertrophy of the left ventricle together with dilatation of its cavity in these patients. PMID- 10476645 TI - [Helicobacter pylori prevalence among patients with gastroduodenal diseases in Kiev]. AB - It is for the first time in Ukraine that a study has been made on the epidemiology of H. pylori infection among patients with disorders of the stomach and duodenum. The diagnosis of the infection was done with the aid of various invasive and noninvasive tests, 13C-urea breathing test, a unique for Ukraine and countries of the Western Europe method of examination included. Ascertained among the above category of patients was wide prevalence of H. pylori, which fact confirms its leading etiological role in disorders of the stomach and duodenum once again. PMID- 10476646 TI - [Pancreatic blood flow function in patients with duodenal peptic ulcer]. AB - The condition was studied of bloodflow in 29 patients with duodenal ulcer, operated on because conservative therapy proved to be ineffective. In patients with uncomplicated duodenal ulcer, disorders of the pancreas bloodflow are at their greatest in the area of the head of the gland at the level of arterioles, with arteriolospasm phenomena being most noticeable. The secured data are to be taken account of in selection of therapeutic measures to be instituted in the patients. PMID- 10476647 TI - [The semiotics of the pain and dyspeptic syndromes in motor disorders of the digestive organs in children and adolescents]. AB - Overall 304 children and adolescents with gastro-duodenal pathology were studied for some aspects of clinical manifestations of the pain and dyspeptic syndromes as related to the character of disordered gastroduodenal motility by making use of techniques of phase polygastroduodenometry. Pathogenetic interrelationship was disclosed of clinical manifestations of the pain and dyspeptic syndromes according to the variant of gastroduodenal dysmotility (dysphasic hyper- or hypomotile dyskinesia of the stomach and duodenum). PMID- 10476648 TI - [The nonspecific reactivity of patients with tuberculosis of the respiratory organs and different haptoglobin phenotypes]. AB - In patients with haptoglobin (Hp) 2-2, the incidence rate of tuberculosis of respiratory organs was found out to be 1.7 times as high as among patients with Hp 2-1 and 1-1; in those persons with aggravated heredity this parameter was 3.4 times as high. Based on the above data, in 414 first diagnosed cases with different phenotype pulmonary tuberculosis a comparative analysis has been done of certain factors of nonspecific bodily reactivity, which showed that in patients with Hp 2-2 versus Hp 2-1 blood levels of total haptoglobin, ceruloplasmin get decreased while total proteolytic activity is noted to be on the increase. The lysozyme concentration is significantly lower than in patients with Hp 2-1 and remains within the normal range. Poorly manifest reaction to the infectious agent of ceruloplasmin, total Hp, lysozyme which have distinct effects and are interrelated with mechanisms of immune defense creates a favourable background in subjects with Hp 2-2 for origination, development and progressing of the tuberculous process. PMID- 10476649 TI - [The characteristic of the initial hemostatic status of patients requiring surgical treatment for lung diseases]. AB - Overall 164 patients were examined, who had been assigned to surgical treatment for nonspecific pyo-inflammatory diseases, tuberculosis, and pulmonary neoplasia, with the purpose of assessing the baseline hemostatic abnormalities. The presence of hypercoagulation and of hyperfibrinogenemia was proved to be the case against the background of low activity of fibrinolysis in all patients but the ratio of the above components and the mechanism of origination of abnormalities differed depending on the kind of the underlying disorder. A differentiated approach is recommended towards selecting corrective means for dealing with hemostatic inadequacies in treating the above-named conditions. PMID- 10476650 TI - [Cytomegalovirus infection and glomerulonephritis]. AB - A total of 56 patients were studied with different forms of glomerulonephritis (GN). Of these, 15 subjects presented with acute, 41--chronic form of GN. The control group was 10 essentially healthy subjects. In the GN patient group, IgG to CMV was identifiable in 68 percent of patients, IgM--in 54 percent; there were only 20 percent of those demonstrating immunoglobulins to CMV among healthy subjects. CMV infection was most common among patients with nephrotic and latent forms of ChGN. Clinically, the infected patients showed a marked proteinuria, hematuria, and imbalance in the system of immunity manifested by a rise in the number of T-helpers, reduction in T-suppressors, IgG, activity of complement and system of phagocytosis. PMID- 10476651 TI - [A sex life and the use of alcoholic drinks]. AB - Illicit sexual relationship and drinking of alcoholic drinks were brought into statistically significant "competitive" relationship in a testing examination of 957 men and women on the basis of the system analysis of the secured findings, which fact has a peculiar effect on both the age of the start of sexual life and the first occurrence of being infected with a venereal disease. Established in the above examination was also the character of correlation between the rate and ways for formation of types of drinking alcoholic drinks and level of education, age, and social status. PMID- 10476652 TI - [The clinico-laboratory criteria of brain death in neurosurgical patients]. AB - With the purpose of developing scientifically substantiated criteria for brain death complex of clinical and physiological, biochemical, physicochemical, and morphological studies was initiated in 55 patients in a critical state because of a dangerous craniocerebral injury, tumours, and vascular affections of the brain. Generalized in the paper are well-known parameters characterizing irreversibility of changes. The following items are to be regarded as the principal criteria for brain death at the present stage of development of medicine: death of truncus cerebri and irreversibility of metabolic disturbances in brain activity. PMID- 10476653 TI - [Catecholamine metabolism in patients with neurological manifestations of spinal osteochondrosis]. AB - Catecholamines content was measured in blood plasma and urine in 104 patients presenting with different neurological syndromes of vertebral osteochondrosis. The results secured were compared to the values of vegetative cardiovascular tests in the same patients. Patients presenting with reflex vegetative muscular and-tonic, reflex vegetovascular manifestations of vertebral osteochondrosis, and those patients presenting with radix symptomatology as well as those patients with reflex neurodystrophic manifestations (though less evident in the latter) were all diagnosed as having developed relatively high values for blood plasma content of norepinephrine at the expense of degeneration of peripheral vegetative nerve endings and reduction of the normal recapture of this mediator from the blood and its urine elimination. Diagnosing of high values for blood plasma and urine content of norepinephrine against the background of a striking inhibition of indices of vegetative cardiovascular tests in patients with vertebrogenic radix symptomatology suggest involvement into the bodily process of compensation and adaptation of suprasegmental vegetative nerve structures. PMID- 10476654 TI - [The antibiotic sensitivity of the tonsillar staphylococcal and streptococcal flora in children who are frequently ill]. AB - The frequency of recovery was studied as was sensitivity to antibiotics of staphylococci and streptococci of the faucial tonsils over the last 7 years in those children with chronic tonsillitis who are in poor health. In always ailing children, it is Staphylococcus aureus that is recovered from the tonsils more often than not, with pathogenic streptococcus being found on more rare occasions. Sensitivity of pathogenic streptococci to benzyl penicillin sodium salt has gotten much lower in recent years. This accounts for a low efficiency of penicillin therapy of streptococcal infection, which fact is to be considered in designing therapeutic measures to be instituted in those children being in poor health. PMID- 10476655 TI - [The determination of the cellular immunity indices in papillomavirus infection of the female genitalia]. AB - The authors studied the impact of HPV infection and HPV-associated cellular intraepithelial neoplasia (CIN) and cancer in situ on indicators of cellular immunity, such as levels of T(CD3+)- and B (CD19+)-lymphocytes as well as those of their subpopulations (CD4+, CD8+, CD3+DR+, CD3-DR+). Female patients presenting with benign states and CIN II as well as those with CIN III and cancer in situ induced by HPV exhibit an apparent decrease in CD3+DR+T-lymphocyte count in their peripheral blood. In benign states and CIN I as well as in CIN III an cancer in situ levels of CD4(+)-cells are noted to be on the decrease while those of CD8+ T-lymphocytes get appreciably increased. Female patients with benign states and CIN I as well as those with CIN III and cancer in situ have a reduction in the index CD4/CD8 compared to the values in controls. The results obtained may have a prognostic significance and will, we believe, help in developing methodological approaches to the immunocorrection of HPV infections and HPV-associated disorders of female genital organs. PMID- 10476656 TI - [The hemodynamic status of patients with infectious myocarditis]. AB - Acute infectious myocarditis is accompanied by disturbances in the contractile function of the left ventricle, its degree being related to the gravity of the disease course. Improvement in the condition of the left ventricular contractile function correlates both with degree of severity of the illness and methodological approaches to the conducted therapy. PMID- 10476657 TI - [Food risk factors and arterial hypertension in a rural population of Chernigov Province]. AB - The prevalence of consumption of foodstuffs in the rural population of Chernigov Province was studied as were aspects of diet among the population depending on the presence of arterial hypertension (AH), for which purpose a special questionnaire was used permitting the objective assessment to be carried out of the character of nutrition. Most prevalent among the rural population is consumption of predominantly animal fat, eggs, sour-milk foodstuffs, sugar, and legumina. In AH patients, a not-rational diet is manifested by a high consumption of saturated fats and inadequate intake of polyunsaturated fatty acids, vitamines, and mineral substance. PMID- 10476658 TI - [Marfan syndrome and pregnancy]. AB - Reported in the paper are three cases of women at reproductive age presenting with Marfan's syndrome. They all underwent urgent surgery because of a threat of rupture of dissecting aneurysm during pregnancy (n = 1). All experienced uneventful recovery. Recommendations are giver as to observation over such patients. The author has reached the conclusion that in case of impending or already developed dissection it is necessary that cesarean section be done on an urgent basis to save life of the mother and fetus alike, with surgical correction of aortic aneurysm and of aortic incompetence being indicated following the cesarean operation. PMID- 10476659 TI - [The pathomorphology and pathogenetic problems of liver involvement in icterohemorrhagic leptospirosis]. AB - Morphologic findings on the liver during different time periods of Leptospira infection suggest the leading role in the pathogenesis of the damaged organ early in the course of the disease of a toxic vascular affection of the microcirculatory bed presenting with hyperpermeability of the vascular wall, interstitial edema, dyscomplexation of the liver crosspieces together with inflow of bile to the sinusoidal capillaries. To a lesser extent jaundice is related to the hepatocyte dystrophic and necrotic changes as evidenced by the absence of marked disturbances in the protein-synthetizing function of the liver and low enzymatic activity. At week 2 to 3 of the illness against the background of diminution of the edema and partial or complete restoration of the liver constitution (especially so in less afflicted peripheral portions of the lobes) jaundice with high bilirubinemia is caused by a toxic affliction of hepatocytes and advancing centrolobular cholestasis. High values for bilirubin might be related not only to grave alternative changes in hepatocytes but to microcirculatory abnormalities as well, which fact accounts for the absence in leptospirosis of a direct relationship of the blood level of bilirubin to the degree of affliction of the hepatic cells. PMID- 10476660 TI - [The use of substitute surfactant therapy in treating acute destructive pneumonia in children]. AB - The article presents results of employment of replacement surfactant therapy with the exogenous surfactant preparation sukrim as treatment of the infiltrative destructive form of acute purulent destructive pneumonia in children. Based on clinical, laboratory, and roentgenological findings a comparative evaluation was done of the course of disease in 20 patients having received substitution surfactant therapy and in 25 those having not been given it. Employment of the preparation suchrim and a short-term course of antibiotic therapy combined in treatment of acute destructive pneumonia in pediatric patients proved to be a highly efficient treatment option for this pathology. PMID- 10476661 TI - [The effect of the ultraviolet irradiation of the blood and of hemosorption on the biochemical signs of endogenous intoxication in asthma patients]. AB - Effects were studied of ultraviolet irradiation of blood and of hemosorption on nonspecific signs of inflammation and endotoxic syndrome such as serum level of medium-weight molecules and percentage of their hydrophobic portion, the content of sialic acids and seromucoid, the binding capacity of blood proteins, and extent of conformational alterations in albumin. It has been ascertained that medium-molecular weight substances are a factor in the pathogenesis of endotoxic syndrome in patients with bronchial asthma, which fact is a reflection of exacerbation of the illness. It is suggested that medium-molecular weight substances might have a part in the decline of the blood protein transport function and in progressing of the conformational alterations in the principal transport protein albumin. Hemosorption and ultraviolet irradiation of blood make for alleviation of endotoxic syndrome and offer advantage of shortening the time the remission supervenes in. The above techniques have direct antiinflammatory and detoxicating effects. PMID- 10476662 TI - [The reasons for the ineffective treatment of patients with newly diagnosed destructive pulmonary tuberculosis]. AB - As many as 350 patients with ineffective treatment of freshly detected destructive pulmonary tuberculosis were analyzed in retrospection. The following four groups of causes of ineffective treatment of these cases were identified: tardy detection and late start of the treatment (49.71%); features of the tuberculous process, the extent to which it has spread in particular; dangerous complications and intercurrent illnesses (66.57%); a not-rational treatment of a patient with tuberculosis (85.14%); untimely discontinuance of treatment (38.86%). In a major proportion of patients these causes tended to be combined, which fact is to be kept in mind in one's daily work for such causes to be cleared away. PMID- 10476663 TI - [The characteristics of the course of acute and chronic glomerulonephritis in adults of different ages]. AB - A total of 846 patients with glomerulonephritis were examined. Of these, 219 were diagnosed as having developed acute glomerulonephritis (AGN), 627 were patients with chronic glomerulonephritis (ChGN). AGN is marked by a debut predominantly in the pubertal period and early adulthood, the incidence of the uric syndrome gets decreased with aging while that of the nephritis syndrome is noted to be on the increase in AGN patients. The course of ChGN depends on the age of the patient having developed the illness: the older the age group that such a patient belongs in, the more unfavourable the disease course is expected to be: developed early in the course of the disease is the hypertonic stage and that of chronic renal insufficiency as well, with the average length of life getting shortened beginning with the onset of the illness. PMID- 10476664 TI - [The clinico-psychopathological characteristics of affective disorders]. AB - Based on the study of 228 depressive patients presenting with various nosological entities of the disorder a detailed clinical and psychopathological characterization was done of depressive disorders. The authors makes out five clinical variants related to features of the pathological process and principal clinical manifestations of the disorder in question. The role of different social and psychological factors in genesis of present-day depression is analyzed. PMID- 10476665 TI - [The structure, severity and dynamics of disability as a consequence of chronic posttraumatic osteomyelitis]. AB - Medical documentation was studied as were experts' reports in 256 patients and invalids. The specific weight of those patients with posttraumatic osteomyelitis assigned to expert medical evaluation was noted to be on the increase (4.5% in 1995 versus 2.7% in 1992), with posttraumatic osteomyelitis of the lower extremity being most prevalent (90.6%). The problem of invalidity secondary to chronic posttraumatic osteomyelitis is still urgent, which fact necessitates developing innovative treatment modalities for dealing with injuries and their complications. PMID- 10476666 TI - [The effect of the parameters of the therapeutic protocol on the efficacy of the adjuvant chemotherapy of breast cancer]. AB - A 5-year relapse-free survival was analyzed in two groups of patients with stage II breast cancer, having received adjuvant chemotherapy (AChTh). The efficiency of adjuvant monochemotherapy with thiophosphamide is inferior to results of employment with the same purpose of six cycles of CMP. The results of AChTh with thiophosphamide appeared to be related to the total dose of the drug. With its course dose of up to 200 mg the value for a 5-yr relapse-free survival is half as much as the efficiency of treatment of those patients having received more than 210 mg of the drug. Similarly, deviations from the requirement of the standard CMP protocol (100-40-600, 6 cycles with a 29-day interval) toward reduction of single doses of cytostatics and diminution of the number of chemotherapy cycles are accompanied by a significant worsening of long-term results of supplementary treatment. Efficiency of AChTh in circumscribed affection of regional lymphatic nodes is much higher than in the presence of four and more lymph-nodal metastases. PMID- 10476667 TI - [The intensive drug treatment of disseminated ovarian ovarian cancer]. AB - Profound endogenous intoxication is a feature of extensive ovarian cancer (OC). In modern oncological gynecology perfection of means and ways of disintoxication is achieved through stepping up of polychemotherapy. This involves a combined use of antioxidants and enterosorption enhancing the efficiency of polychemotherapy, improving OC patients' quality of life. PMID- 10476668 TI - [A case of the successful use of Motilium in treating reflux disease against a background of the hypothalamic syndrome of traumatic origin]. PMID- 10476669 TI - [Iron-deficiency anemia as a hematological mask of myxedema]. AB - An atypical course of myxedema manifested by iron-deficiency anemia is described that proved to be a diagnostic challenge. Pathogenetic mechanisms of origination are analyzed. PMID- 10476670 TI - [The efficacy of the multienzyme preparation Wobe-Mugos E in lymphoproliferative diseases]. AB - The drug Wobe-Mugos E was prescribed to patients with chronic lymphoid leukosis and multiple myeloma during and following the administration of chemotherapy treatment, 3 tablets on a three-times daily schedule 30 to 45 minutes before meals for 2 to 4 weeks. A positive effect was manifested by improvement in morphofunctional properties of erythrocytes, detoxicating and antioxidant effects. There were no cases of intolerability or side effects of the drug in the examined patients. PMID- 10476671 TI - [The pharmacological activity and clinical use of oxytocin and melanostatin]. PMID- 10476672 TI - [The clinico-laboratory effects of acupuncture in patients with glomerulonephritis]. AB - As many as 104 patients with different clinical forms and variants of glomerulonephritis were examined and treated with acupuncture. This mode of treatment was found out to positively influence the level of hemoglobin, ESR, eosinophilia, making for diminution of proteinuria and erythrocyturia, returning hemostasis and blood biochemical spectrum (proteinogrammes, levels of cholesterol, uric acid) to normal, with the level of arterial pressure being normalized in patients with mild hypertension or reduced in those with moderately severe to severe hypertension. Acupuncture is recommended as a monotherapeutic modality in the treatment of uric syndrome with a hematuric component in a combined treatment of nephrotic syndrome and hypertension, which attempt will, we believe, help in preventing side effects of the drugs employed. PMID- 10476673 TI - [The functional characteristics of the immune system, of lipid peroxidation and of the endogenous antioxidant system in miners working in deep coal mines]. AB - A study was made of the condition of the immune status and lipid peroxidation in 77 essentially healthy underground miners of a coal mine that showed a decline in cellular as well as humoral links of immunity and increase in amounts of circulating immune complexes. Moreover, the miners exhibited a rise in conjugated dienes, malonic dialdehyde, and catalase. The workers were divided into groups depending on how difficult were jobs they were assigned to. A relatedness was found out of enhancement of processes of lipid peroxidation and activity of antienzymatic defence to strenuousness of work performed. PMID- 10476674 TI - [Peripheral autonomic insufficiency in workers who sustain the chronic effect of an occupational vibration-noise factor]. AB - The development was studied of vegetative disorders in workers under chronic exposure to occupational vibration and noise. In the workmen, the segmentary apparatus of the vegetative nervous system (VNS) gets affected, with the peripheral vegetative insufficiency syndrome being developed. During the early stages of occupational contact with a vibronoise factor there develops an affection of the sympathetic portion of VNS but the longer the service duration the greater is the share of parasympathetic disorders. One of the most efficient ways for preventing a vibronoise affection of VNS are well-done preliminary and regular health checks, observance of sanitary-and-health labour standards, and introduction of innovative technological ways and means designed to offer defence against vibration and noise. PMID- 10476675 TI - [The principles of medical care development during the transition to mandatory medical insurance]. PMID- 10476676 TI - [The organization of the work of the mobile emergency medical care teams at medical first aide stations]. PMID- 10476677 TI - Separation of dorsal and ventral dopaminergic neurons from embryonic rat mesencephalon by buoyant density fractionation: disassembling pattern in the ventral midbrain. AB - The dopaminergic neurons of the ventral mesencephalon, though physically mixed with non-dopamine neurons, are organized into dorsal and ventral 'tiers' with regard to their ontogeny, efferent projections and their relative position in the various mesencephalic sub-nuclei. We have employed buoyant density fractionation to separate the dopaminergic neurons of the two compartments and compare their subsequent phenotype development with respect to their expression of the gene encoding tyrosine hydroxylase, the rate-limiting enzyme in the catecholamine biosynthetic pathway. Using immunocytochemistry, separately and combined with in situ hybridization, we demonstrate here that sedimentation of cell suspensions from E19 rat ventral mesencephalon on 5-step Percoll gradients produces cell fractions enriched in ventral and dorsal tier DA neurons, respectively. PMID- 10476678 TI - Probing intracellular dynamics in living cells with near-field optics. AB - Near-field optics (NFO) overcomes the diffraction limit of light microscopes and permits visualization of single molecules. However, despite numerous applications of NFO in the physical sciences, there is still a paucity of applications in the neurosciences. In this work, the authors have developed NFO probes to image intracellular dynamic processes in living cells. This is the first time a NFO probe has been inserted inside a living cell to deliver light to a spatially controlled region for optical measurements and to record cellular responses to external stimuli. Two different optical detection systems (CCD camera and avalanche photon detection) were developed to monitor cellular responses to drug administration in two different cell types. NG108-15 neuroblastoma cells and vascular smooth muscle cells (VSMC) were penetrated with NFO probes. Intracellular Ca2+ increases post drug stimulation were detected by NFO probes. The cells were loaded with either fura-2/AM or fluo-3/AM calcium dyes. VSMC were stimulated with angiotensin II, resulting in a precise area of intracellular Ca2+ increase. Different response profiles of Ca2+ increases were observed after ionomycin and bradykinin administration in NG108-15 cells. Responsive heterogeneities due to ionomycin among different cells of the same type were recorded. The results show that NFO probes make possible real-time visualization of intracellular events. With refinement, intracellular NFO probes offer the potential of probing cell function with fast temporal and excellent spatial resolutions. PMID- 10476679 TI - In vivo predegeneration of peripheral nerves: an effective technique to obtain activated Schwann cells for nerve conduits. AB - In vivo predegeneration of peripheral nerves is presented as a convenient and effective method to obtain activated Schwann cells and an enhanced cell yield following in vitro cultivation. The experiments conducted in rats were aimed at clinical use in gaining Schwann cell suspensions for filling artificial conduits in order to bridge peripheral nerve gaps. The rat sciatic nerve used as a model was transected distally to the spinal ganglia. Predegeneration in vivo was allowed to take place for 1, 2, 3 and 4 days and up to 1, 2 and 3 weeks. The nerve was then resected and prepared for cell cultivation. Schwann cells cultivated from the contralateral untreated nerve served as control. Immunostaining for S100, nerve growth factor receptor and the adhesion molecules N-cadherin and L1 was used to characterize the general state of the cultures. Viability was assessed by fluorescein fluorescence staining, and the proliferation index was determined by bromodeoxyuridine-DNA incorporation. The Schwann cells from predegenerated nerves revealed an increased proliferation rate compared to the control, whereas fibroblast contamination was decreased. Best results were obtained 1 week after predegeneration. PMID- 10476680 TI - A submersion method to induce hypoxic damage in organotypic hippocampal cultures. AB - Organotypic brain slices cultured on semi-porous membranes is an increasingly popular in vitro preparation for studying mechanisms of ischemic brain damage. To model in vivo hypoxia, cultured brain slices are exposed to anaerobic atmosphere by placing them into a special incubator. This requirement limits the use of in vitro ischemic models to highly specialized laboratories. Here, we describe a simple method that reproduces hypoxic injury, where cultured hippocampal slices are submerged into glucose-free deoxygenated medium for 1 h. The extent and distribution of hippocampal neuronal loss obtained with this treatment resembled that caused by hypoxia in living tissue in situ, i.e. CA1 pyramidal cell layer was most vulnerable and dentate granular cell layer was least susceptible to hypoxia as measured with fluorescence of the viability marker propidium iodide (PI). Electrophysiologic functional impairment determined by field recordings of CA1 pyramidal neurones temporally coincided with the extent of neuronal death. In addition, known neuroprotective treatments, such as hypothermia and phenytoin application ameliorated neuronal damage in a pattern similar to previously published reports. Therefore, the present in vitro model of ischemia is simple, reliable and of low cost. It is well suited for short and long-term studies of the mechanisms of hypoxic brain damage. PMID- 10476682 TI - Measurement of the limiting equivalent conductivities and mobilities of the most prevalent ionic species of EGTA (EGTA2- and EGTA3-) for use in electrophysiological experiments. AB - In many experimental biological situations, chelating agents like EGTA (ethylene glycol-bis-(beta-amino-ethyl ether) N,N,N',N'-tetra-acetic acid) are commonly used to control or suppress the concentration of divalent ions like Ca2+. The evaluation of liquid junction potentials in electrophysiological measurements, and particularly in patch-clamp situations, requires information about the ions within the solution. Where there is a significant concentration of EGTA present, it is necessary to know the values of the relative mobility of at least the most predominant ionic species of EGTA in order to complete these calculations. EGTA, with four negative charges with different pKas, can therefore exist as four differently charged ions in solution (EGTA-, EGTA2-, EGTA3- and EGTA4-) or as uncharged, although between pH 5.5 and 8 it is almost exclusively EGTA2-. We have measured limiting equivalent conductivities of the most common ionic forms of EGTA (EGTA2- and EGTA3-) encountered at physiological pHs. These were 35.9 +/- 0.7 and 56 +/- 2.5 S cm2 equiv(-1) respectively. Their mobilities relative to K+ were 0.24 +/- 0.01 for EGTA2- and 0.25 +/- 0.01 for EGTA3-. Thus for typical electrophysiological solutions, the contribution of EGTA to the liquid junction potential should be small (e.g. approximately 0.4 mV). PMID- 10476681 TI - Visualization of synaptic specializations in live C. elegans with synaptic vesicle protein-GFP fusions. AB - Synaptic specializations are difficult to visualize at the light microscope level in living preparations. To circumvent this problem, synaptic vesicle proteins were fused to green fluorescent protein (GFP) and expressed in C. elegans neurons. C. elegans synaptobrevin-GFP and synaptogyrin-GFP fusion proteins were observed to target to synaptic sites. This localization allowed the visualization of synaptic specializations in living animals with light microscopy. Restricted expression of synaptobrevin-GFP fusions in subsets of neurons enables the visualization of individual presynaptic varicosities. The cell biology underlying the sorting of synaptic vesicle proteins, trafficking of vesicles to terminals, and the development of presynaptic specializations is now more amenable to forward genetic analysis using these synaptic markers. PMID- 10476683 TI - Targeted expression of human CuZn superoxide dismutase gene in mouse central nervous system. AB - Copper zinc superoxide dismutase (CuZnSOD) is an important enzyme for the detoxification of reactive oxygen species. Particularly in the central nervous system (CNS), reactive oxygen species are often associated with acute brain injuries and chronic neurodegeneration. It has been demonstrated in vivo that there is an inverse correlation between CuZnSOD activity and neuronal death after acute brain injury. To further understand the protective role of CuZnSOD upon neurons, we have generated transgenic mouse lines with targeted expression of the human CuZnSOD gene (SOD1) that is driven by a rat neuron-specific enolase gene promoter in neurons of the CNS. The transgenic SOD1 expression was restricted to the CNS identified by reverse transcriptase polymerase chain reaction and SOD gel electrophoresis assays. The CuZnSOD activity was significantly increased in the brain stem of the transgenic mice. Immunostaining of human CuZnSOD activity showed that Purkinje cells in the cerebellar cortex were the most intensely stained neurons in the CNS of the transgenic mice. PMID- 10476684 TI - Functional activation of punch-cultured magnocellular neuroendocrine cells by glutamate receptor subtypes. AB - To provide a simple means to isolate and study the cellular functions of small groups of neurons, we developed a modified 'punch' culture procedure that facilitates acute and long-term in vitro physiological studies. Primary 'punch' cultures of magnocellular neuroendocrine cells (MNCs) from the supraoptic nucleus (SON) were established and the basic physiological effects of subtype-specific glutamate receptor agonists were characterized. MNCs from the punch cultures established a mature morphology in culture with extensive outgrowth of axons and varicosities. After 8 days, a single cultured SON punch produced an average of 10.0 +/- 2.1 pg AVP and contained an average of 222 +/- 53 vasopressin neurophysin immunoreactive cells. Patch clamp recordings from MNCs demonstrated the presence of N-methyl-D-aspartate (NMDA)-sensitive and DL, alpha-amino-3 hydroxy-5-methylisoxazole propionic acid (AMPA)-receptors. Stimulation of metabotropic receptors with 1S,3R ACPD induced acute or gradual increases in intracellular calcium. NMDA, AMPA and metabotropic receptors all contributed to the secretion of vasopressin from the punch cultures with an agonist rank order potency of: NMDA (10 microM) > AMPA (1 microM) = 1S,3R ACPD (100 microM) > kainate (10 microM). This culture preparation should be useful for cellular studies of small groups of neuroendocrine and other cells. PMID- 10476685 TI - Measurement of neurotrophin 4/5 in rat tissues by a sensitive immunoassay. AB - Neurotrophin 4/5 (NT4/5) is a member of the neurotrophin family known to exert survival and other effects on a variety of neurons including those within the motor, sensory and central populations. Although mRNA(NT4/5) has been found in various effector tissues of the rat and human, the concentration of NT4/5 protein in tissues has not been reported previously due to lack of suitable methodology. We present here a quantitative two-site enzyme-linked immunosorbent assay for the estimation of NT4/5 in pre- and postnatal rat tissues. The assay was performed using a combination of polyclonal and monoclonal antibodies to recombinant human NT4/5. Tissue samples were extracted at neutral pH. Results show that the assay is highly specific for NT4/5 with a sensitivity of 1 pg/ml, and reproducible with intra- and inter-assay variation coefficients of 3.0 and 6.3%, respectively. NT4/5 was found in most embryonic tissues examined at gestation day 17 and 21, but was rarely detectable in postnatal tissues, with the notable exception of the testis. The availability of an immunoassay for the estimation of NT4/5 protein in rat tissues should contribute to the understanding of the physiology of this little understood neurotrophic factor. PMID- 10476686 TI - Differential sensitivities of the two visual pathways of the chick to labelling by fluorescent retrograde tracers. AB - This study investigates the neurone structure-specific differences of sensitivities of fluorescent tracers. The tracers were used for retrograde labelling of contralateral projections in the two visual pathways of the chick. Rhodamine B Isothiocyanate (RITC), Fluorogold (FG) and True blue (TB) were injected into either the visual Wulst (thalamofugal pathway) or the nucleus rotundus (Rt; tectofugal pathway) and the retrogradely labelled neurones in the nucleus geniculatus lateralis pars dorsalis (GLd) or the optic tectum, respectively, were counted. Differential retrograde labelling in the two pathways was observed. In the thalamofugal pathway, both the contralateral and ipsilateral GLd cells were labelled by all three tracers (RITC, FG and TB). However, in the tectofugal pathway, whereas RITC labelled both the ipsilateral and contralateral tectal neurones, FG or TB labelled effectively only the ipsilateral tectal neurones. It was clear that FG and TB were taken up by the nerve endings and transported part-way along the axon but failed to be transported to the cell bodies of the contralateral tectal neurones. In addition, red beads and green beads were also injected into Rt and the differential labelling was also observed. Red beads labelled both ipsilateral and contralateral tectal neurones but green beads labelled only the ipsilateral tectal neurones. Since the contralateral tectal projections consist of divergent axon collaterals, the present study suggests that various retrograde tracers are not transported in these axon collaterals to label cell bodies. The contralaterally projecting neurones in the thalamofugal pathway are not axon collaterals and they were labelled by all of the tracers used. PMID- 10476687 TI - Progesterone-induced blocking factor inhibits degranulation of natural killer cells. AB - PROBLEM: During the first trimester of pregnancy, nonclassical (CD3-, CD56+, CD16 , perforin [P]bright+) natural killer (NK) cells comprise the major decidual lymphocyte population. These cells, in spite of their high perforin content, exert a low cytolytic activity. Peripheral blood lymphocytes of healthy pregnant women produce progesterone-induced blocking factor (PIBF), which inhibits NK activity. PIBF-producing cells are likely to be present in decidua and might contribute to low decidual NK activity. METHOD OF STUDY: Decidual cells obtained from elective pregnancy termination were double labeled for CD56 and PIBF. We tested the effect of PIBF on perforin liberation by activated peripheral blood NK cells. RESULTS: Sixty percent of decidual lymphocytes were CD56 + and expressed PIBF at the same time. PIBF-treated and untreated peripheral blood NK cells were incubated with K-562 cells, and perforin content of target conjugated NK cells was detected with immunocytochemistry. PIBF treatment of peripheral blood lymphocytes significantly reduced lysis of K-562 cells. Among target bound lymphocytes in PIBF-treated samples, we found a significantly (P < 0.01) higher rate of P+ cells than in untreated samples. CONCLUSIONS: These data suggest that PIBF inhibits cytotoxicity of NK cells via a block of degranulation, and since decidual NK cells are PIBF+, it cannot be ruled out that this effect of PIBF contributes to low decidual NK activity. PMID- 10476688 TI - Decidual macrophages are the population of decidual adherent cells which regulates perforin expression in cytolytic cells. AB - PROBLEM: We have shown that addition of decidual adherent cells (DAC) to the culture of decidual lymphocytes (DL) prevents the downregulation of perforin expression in these cells. Because DAC are a mixture of various cell populations, the aim is to analyze immunophenotypic characteristics of DAC and to determine which cell population is involved in the regulation of perforin expression. METHOD OF STUDY: First trimester pregnancy decidual cells were obtained by enzymatic tissue digestion. Decidual cells and peripheral blood lymphocytes (PBL) were centrifuged on Ficoll-Hypaque density gradient and cultured overnight to obtain adherent cells, which were analyzed by flow cytometry and immunocytochemically. RESULTS: Almost all peripheral blood adherent cells (PBAC) (ca 90%) expressed monocyte/macrophage markers but only 10-20% of DAC. The rest of DAC expressed markers of stromal cells. HLA-DR depleted population of DAC (stromal cells only) could not prevent downregulation of perforin expression in cultured DL and PBL. CONCLUSION: Decidual macrophages are involved in the regulation of perforin expression in DL. PMID- 10476689 TI - The role of gamma/delta T-cell receptor-positive cells in pregnancy: part II. AB - PROBLEM: We have previously demonstrated a significantly increased ratio of gamma/delta T-cell receptor (TCR)-positive progesterone receptor(PR)-positive cells in the peripheral blood of healthy pregnant women compared to that of recurrent aborters or non-pregnant individuals. Treatment of pregnancy lymphocytes with a pan anti-gamma/delta TCR antibody inhibits progesterone induced blocking factor (PIBF) production, increases natural killer (NK) activity, and alters the cytokine profile. The present study was aimed at investigating the role of the different gamma/delta subpopulations in these phenomena. METHOD OF STUDY: Peripheral blood lymphocytes from healthy pregnant women were incubated with either anti-gamma1.4 and delta1, or anti-gamma9 and delta2 antibodies. The effect of these treatments on PR induction and interleukin (IL)-10 and IL-12 expression were tested by immunocytochemistry. NK activity of anti-gamma/delta treated lymphocytes was also determined. RESULTS: In peripheral blood of healthy pregnant women, the most frequently occurring chain combination was gamma1.4/delta1, whereas in recurrent aborters, the gamma9/delta2 combination was predominant. Treatment of normal pregnancy lymphocytes with a mixture of gamma1.4 and delta1 antibodies resulted in a significantly reduced NK activity and increased PR and IL-10 expression, whereas treatment with a mixture of gamma9 and delta2 antibodies significantly reduced IL-10 production and slightly increased IL-12 production and NK activity. These data suggest the presence of two functionally distinct subpopulations in the peripheral blood of pregnant women. PMID- 10476690 TI - Apoptotic nuclei within the uterine decidua of first trimester pregnancy arise from CD45 positive leukocytes. AB - PROBLEM: The Fas/Fas-ligand (FasL) system is discussed to be one of the main components responsible for the formation and maintenance of an immune privilege. As we were able to detect FasL on fetal trophoblast cells infiltrating the maternal uterine tissue and Fas on decidual leukocytes (Hammer et al., Am J Reprod Immunol, 1999; 41: 41-51), we investigated whether apoptosis occurs in the maternal decidua during the first trimester of pregnancy. METHOD OF STUDY: A TUNEL-assay was performed in combination with immunofluorescence to detect and characterize cells undergoing apoptosis. RESULTS: Apoptotic nuclei mainly arising from CD45+ leukocytes could be detected in the trophoblast-invaded decidua basalis as well as in the non-invaded decidua parietalis. However, while in the decidua parietalis the apoptotic leukocytes are scattered within the whole functional stroma, in the decidua basalis they are concentrated in the area around the so-called "necrotic foci" and especially in the primary feto-maternal contact zone. where trophoblast cells from the cell columns begin to invade the maternal tissue. CONCLUSION: Because of the irregular distribution of apoptotic CD45+ leukocytes in the decidua basalis we think that some of the maternal leukocytes are activated and killed by fetal trophoblast cells which express FasL. PMID- 10476691 TI - Progress in characterization of pre-implantation factor in embryo cultures and in vivo. AB - PROBLEM: Pre-implantation factor (PIF), a small, embryo-derived peptide is detected in the maternal serum prior to implantation and is associated with successful pregnancy outcome. However, its identity is not known. METHOD OF STUDY: PIF was isolated from mouse embryo conditioned media and from pregnant porcine sera, using high-performance liquid chromatography (HPLC) followed by mass spectrometry. RESULTS: Conditioned culture media was separated by gel filtration chromatography followed by reversed phase chromatography. At each step, PIF activity was determined by the lymphocyte/platelet binding autorosette assay (LPBA). Mass spectrometry yielded a single peak with a mass of 1300 Da. The peptide is, however, present in very low concentrations (fM), which has so far precluded complete identification. Pregnant porcine sera that exhibit potent PIF activity were deproteinated by acetone and further fractionated by reversed phase HPLC. Active fractions contain peptides of molecular masses 523 and 551 Da. CONCLUSION: PIF, likely to be peptides, represents a novel substance related to pregnancy initiation and maintenance. PMID- 10476692 TI - Cytokines and the regulation of apoptosis in reproductive tissues: a review. AB - PROBLEM: To determine the role of apoptosis-regulating genes bax and bcl-2 in reproduction. METHOD OF STUDY: Review of literature and current data. RESULTS: The bcl-2 family of apoptotic regulatory gene products interact and form dimers of anti- and pro-apoptotic proteins (e.g., bcl-2 and bax respectively), the ratio of which determines cell death or survival. Menses is associated with increased apoptosis in the glands, a change in bcl-2:bax ratio and increased levels of the pro-apoptotic cytokine TNFalpha. Apoptosis occurs in all placental cell types and increases from first to third trimester. Placental apoptosis is induced by TNFalpha in vitro and increased levels in utero characterize most failing pregnancies, intra-uterine growth restriction (IUGR) and labour. An increased bcl 2:bax ratio and apoptosis in the syncytiotrophoblast characterizes failing first trimester pregnancies. Apoptosis in the syncytiotrophoblast is also associated with IUGR. In a rat model, maternal vitamin A deficiency perturbs fetal development. This is associated with a placental infiltrate of TNFalpha positive neutrophils (day 20) and increased placental apoptosis in areas of infiltration. A similar infiltrate occurs in a mouse model of early pregnancy loss. In the fetal membranes, clusters of bcl-2 negative chorion trophoblast cells undergo apoptosis. This may allow passage of myometrial stimulatory factors that induce labour. CONCLUSION: The ratio of bcl-2:bax is crucial in the regulation of apoptosis, particularly in the human placenta. Changes in trophoblast apoptosis characterize (1) early pregnancy failure, (2) IUGR and (3) pre-term and term labour. Regardless of gestational age, TNFalpha plays a major role in the induction of placental apoptosis. PMID- 10476693 TI - The possible role of classical human leukocyte antigens in recurrent miscarriage. AB - PROBLEM: If immunological factors play a role in the pathogenesis of recurrent miscarriage (RM), it is likely that associations between alleles of classical human leukocyte antigen (HLA) genes and RM exist. Our aim was to investigate HLA C alleles in RM couples and HLA-DR and -DQ polymorphism in women with unexplained RM. METHOD OF STUDY: HLA-C alleles were investigated in 35 RM and 30 control couples and HLA-DR and -DQ allogenotypes were investigated in 234 RM patients and 360 controls. All HLA investigations were undertaken by DNA based methods. RESULTS: We found no difference between the RM and control couples in the degree of paternal incompatibility for maternal HLA-C alleles and the distribution of the two HLA-C supertypic specificities that are recognized differently by p58 killer cell inhibitory receptor (KIR) positive natural killer (NK) cells was similar in the two groups. In 97 women with at least four previous miscarriages, significantly higher frequencies of the HLA-DR1,DQ5 and -DR3,DQ2 haplotypes were found compared with 360 controls (P < 0.05 after correction for multiple comparisons). Among 94 RM patients followed prospectively, those with HLA-DR1 and/or -DR3 had a 62% miscarriage rate compared with only 29% among those without these alleles (P < 0.05). A large family study indicated that HLA-DR1 and/or -DR3 positive sisters and brothers' wives of probands with RM had an odds ratio of 5.0 for miscarrying their pregnancies compared with corresponding HLA-DR1 and -DR3 negative relatives. Finally, a meta-analysis of relevant studies based on a MEDLINE search showed that HLA-DR1, -DR3, and -DR4 were significantly increased in Caucasian women with RM. CONCLUSIONS: HLA-DR1, -DR3, and maybe -DR4 show association to RM in Caucasian women whereas no association to classical HLA class I genes including HLA-C can be detected in RM couples. The mechanism by which class II alleles confer susceptibility to RM might be by predisposing to hypersecretion of certain cytokines, e.g., tumor necrosis factor (TNF)-alpha at the feto-maternal interface. PMID- 10476694 TI - Inhibition of trophoblast adhesion to endothelial cells by the sera of women with recurrent spontaneous abortions. AB - PROBLEM: May anti-phospholipid or other autoantibodies interfere with trophoblast endothelial cells interaction in women with unexplained pregnancy losses? METHODS OF STUDY: The sera of 72 women with recurrent spontaneous abortions (RSA) containing antibodies to endothelial cells (28), trophoblast (14), and cardiolipin (10) or lacking antibodies (25), and 26 controls were examined in an inhibition assay of trophoblast adhesion to endothelial cells using an ELISA based on the recognition of trophoblast by antibodies to cytokeratin. RESULTS: Adhesion of trophoblast to endothelial cells was time- and dose-dependent. Patients and control sera inhibited trophoblast adhesion with mean values of 37% and 7%, respectively. Inhibition above 2SD of the mean control value was still observed in 58% of the patients sera and 8% of the control sera. Sera containing antibodies to endothelial cells had higher inhibitory effect (38%) than those with antibodies to trophoblast (23%) and cardiolipin (28%) or lacking antibodies (26%). CONCLUSIONS: Antibodies and other undefined factors in the sera of women with RSA inhibit adhesion of trophoblast to endothelial cells. PMID- 10476695 TI - TGF beta 2 mRNA expression and pregnancy failure in mice. AB - PROBLEM: We describe here a pattern of transforming growth factor (TGF) beta2 mRNA expression at the fetomaternal interface in mice with high rate of resorptions as well as its expression following maternal immunopotentiation. METHOD OF STUDY: TGF beta 2 mRNA expression was evaluated in the uteroplacental units of mice with spontaneous (CBA/J x DBA/2J mouse combination) or cyclophosphamide (CP)-induced pregnancy loss. The effect of immunopotentiation on TGF beta 2 mRNA expression was determined in CP-treated females who underwent nonspecific immunostimulation with xenogeneic (rat) leukocytes. A quantitative analysis of TGF beta 2 mRNA level was performed using RNase protection assay. Distribution of TGF beta 2 mRNA transcripts at the fetomaternal interface was studied by in situ hybridization analysis. RESULTS: RNase protection analysis revealed four TGF beta 2 specific mRNA forms (330, 270, 230, and 170 bp) in the uteroplacental units of mice with either normal or decreased reproductive performance. A significant decrease (about 50%) in the level of TGF beta 2 mRNA was registered in the uteroplacental unit of mice with pregnancy loss as compared to the control mice. TGF beta 2 transcripts were abundant in the uterine epithelium and stroma. A specific hybridization signal was detected also in metrial gland cells and it was found to be substantially lower in CP-treated as compared to intact mice. In the resorbing uteroplacental unit, the expression of TGF beta 2 mRNA was completely lost in the uterine epithelium, and the number of TGF beta 2 mRNA-positive metrial gland cells was lower as compared to the control. Immunopotentiation decreased the resorption rate in mice with CP-induced pregnancy loss and caused a dramatic increase in TGF beta 2 mRNA expression: the level of TGF beta 2 mRNA was found to be higher by 2.0-3.2 fold in the uteroplacental unit of immunized as compared to nonimmunized CP-treated mice. CONCLUSIONS: These data suggest that distortion of TGF beta 2 expression at the fetomaternal interface may be associated with pregnancy failure. It seems that beneficial effect of maternal immunostimulation may at least partly be due to the strong increase in TGF beta 2 mRNA expression at the fetomaternal interface. PMID- 10476696 TI - Reactions to market study executive summary. PMID- 10476697 TI - Reactions to market study executive summary. PMID- 10476698 TI - Reactions to market study executive summary. PMID- 10476699 TI - Reactions to market study executive summary. PMID- 10476700 TI - Feline vaccination protocol needs further scrutiny. PMID- 10476701 TI - Questions validity of equine vaccine field studies. PMID- 10476702 TI - The licensure catch-22. PMID- 10476703 TI - Concerned about drug use in food animals. PMID- 10476704 TI - What is your diagnosis? Extensive alveolar pattern in the caudodorsal and caudoventral aspects of the right and left lungs. PMID- 10476705 TI - Age-related changes to nutritional requirements and digestive function in adult dogs and cats. PMID- 10476706 TI - Effects of furosemide on athletic performance and exercise-induced pulmonary hemorrhage in horses. PMID- 10476707 TI - Evolution of the law on liability for sale of diseased animals. PMID- 10476708 TI - Estimation of the proportions of dogs and cats that are surgically sterilized. AB - OBJECTIVE: To determine an estimate of the proportions of dogs and cats in Texas that are surgically sterilized and whether those proportions differed according to species and sex of the animal, level of responsibility of the owner, or geographic location. DESIGN: Cross-sectional study. ANIMALS: 43,831 dogs and cats > or = 6 months old. PROCEDURE: Information on sterilization rates was provided by 14 licensing agencies and 16 animal shelters in diverse regions of Texas. Univariate and multivariate analyses were used to compare sterilization rates among subpopulations of animals (dogs vs cats, males vs females, sheltered vs licensed, rural vs urban location). RESULTS: Overall, 12,893 (29.4%) of the animals (26.9% of dogs and 32.6% of cats) were sterilized. Proportions of animals sterilized were significantly different among subpopulations. CONCLUSIONS AND CLINICAL RELEVANCE: Although the cause of pet overpopulation is multifaceted, failure of owners to spay and castrate their animals is a major contributing factor. Significant differences in sterilization rates among subpopulations of dogs and cats suggest that organizations encouraging spaying and castration should use motivational techniques specific for the pet owners they are targeting. PMID- 10476709 TI - Assessment of degree of oxidative stress and antioxidant concentrations in dogs with idiopathic dilated cardiomyopathy. AB - OBJECTIVE: To assess degree of oxidative stress and antioxidant concentrations in dogs with idiopathic dilated cardiomyopathy (IDCM). DESIGN: Prospective study. ANIMALS: 18 dogs with IDCM and 16 healthy control dogs. PROCEDURE: Concentrations of malondialdehyde (an indicator of oxidative stress); vitamins A, C, and E; glutathione peroxidase; and superoxide dismutase were measured. RESULTS: Glutathione peroxidase concentration was significantly increased in dogs with IDCM, compared with control dogs. Vitamin A and superoxide dismutase concentrations were not significantly different between groups. A negative correlation was found between disease severity and plasma vitamin E concentration. Disease severity was not correlated with concentrations of other antioxidants. Medications did not significantly affect oxidant or antioxidant concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: The change in glutathione peroxidase concentration and the correlation between vitamin E concentration and disease severity suggest that the oxidant-antioxidant system may play a role in development of IDCM. PMID- 10476710 TI - Pericardial effusion and cardiac tamponade secondary to brodifacoum toxicosis in a dog. AB - Hemorrhage resulting from ingestion of anticoagulant rodenticides may be evident at any traumatized site or in any body cavity. It is important for clinicians to include coagulopathies among the differential diagnoses for pericardial effusion and to evaluate clotting function before routine pericardiocentesis is performed. PMID- 10476711 TI - Eosinophilic dermatitis with edema in nine dogs, compared with eosinophilic cellulitis in humans. AB - A unique eosinophilic dermatitis with edema in dogs is characterized by extremely erythematous coalescing macules and plaques with associated edema, and is similar to eosinophilic cellulitis (Wells' syndrome) in humans. Histopathologic features include a profound eosinophilic dermal infiltrate, focal areas of collagen fiber degeneration surrounded by eosinophils (flame figures), dilated vessels, and dermal edema. Etiopathogenesis is unknown, but a hypersensitivity reaction to medications, arthropod bites, or other foreign antigens is suspected. PMID- 10476712 TI - Results of physical inspection before races and race-related characteristics and their association with musculoskeletal injuries in Thoroughbreds during races. AB - OBJECTIVE: To estimate the relative risk of injury among horses deemed to be at increased risk of injury on the basis of prerace physical inspection findings and to examine the association of injury during races with race-related characteristics. DESIGN: Cohort study. ANIMALS: 2,187 Thoroughbred horses that started 3,227 races in Kentucky. PROCEDURE: All race starts for which a horse was deemed to be at increased risk of injury on the basis of prerace physical inspection findings and a random sample of race starts for which horses were not deemed at increased risk of injury were included in the study. Findings of prerace physical inspection, race-related characteristics, and outcome of the race (race results and whether the horse incurred an injury) were recorded for each race start. Race starts in which a horse incurred an injury during a race were compared with race starts in which injuries were not incurred to identify factors associated with injury during races. RESULTS: Abnormality of the suspensory ligament of the forelimbs detected during prerace physical inspection, racetrack, class of race (claiming race < or = $25,000 vs other classes), and distance of race (< 7 furlongs vs other distances) were significantly associated with increased risk of injury. CONCLUSIONS AND CLINICAL RELEVANCE: Prerace physical inspection findings, particularly abnormalities of the suspensory ligament, may be used to identify horses at increased risk of injury during races. Rate of injury differed among racetracks, and horses in certain types of races (lower-priced claiming races and races of shorter distance) may be at increased risk of injury during races. PMID- 10476713 TI - Frequency of musculoskeletal injuries and risk factors associated with injuries incurred in Quarter Horses during races. AB - OBJECTIVE: To determine the frequency and anatomic location of musculoskeletal injuries incurred by Quarter Horses during races and to compare data from injured horses and matched control horses. DESIGN: Matched case-control study. ANIMALS: 97 Quarter Horses that sustained a musculoskeletal injury during races and 291 horses from the same races that were not injured. PROCEDURE: Data examined included racing history, race-entrant characteristics, racing events determined by analysis of videotapes of races, and, when performed, results of prerace physical inspections. Data for injured horses were compared with data for control horses, using conditional logistic regression. RESULTS: Incidence of a catastrophic injury among Quarter Horses during races was approximately 0.8/1,000 race starts, whereas incidence of musculoskeletal injury during racing was approximately 2.2/1,000 race starts. Odds of musculoskeletal injury were approximately 8 times greater among horses assessed to be at increased risk of injury on the basis of results of prerace physical inspection than for horses not considered to be at increased risk of injury. Evidence was lacking that 2-year old horses were at increased risk of injury or that sex influenced the risk of injury among Quarter Horses during races. CONCLUSIONS AND CLINICAL RELEVANCE: Incidence of racing injury among Quarter Horses appears to be lower than that observed among Thoroughbreds. Regulatory veterinarians can identify horses at increased risk of injury on the basis of prerace physical inspection, indicating that these inspections could be used to reduce the risk of injury during races. PMID- 10476715 TI - Chyloperitoneum and abdominal adhesions in a miniature horse. AB - Chyloperitoneum is a potential cause of colic in horses. Although rare in horses, chyloperitoneum can develop secondary to tearing of mesenteric adhesions and rupture of mesenteric lymphatic vessels. The prognosis for horses with chyloperitoneum depends on the underlying cause and the response to treatment. PMID- 10476714 TI - Effect of furosemide on performance of Thoroughbreds racing in the United States and Canada. AB - OBJECTIVE: To determine the effect of furosemide on performance of Thoroughbreds racing on dirt surfaces at tracks in the United States and Canada. DESIGN: Cross sectional study. ANIMALS: All Thoroughbreds (n = 22,589) that finished a race on dirt surfaces at tracks in the United States and Canada between June 28 and July 13, 1997 in jurisdictions that allowed the use of furosemide. PROCEDURE: Race records were analyzed by use of multivariable ANOVA procedures and logistic regression analyses to determine the effect of furosemide on estimated 6-furlong race time, estimated racing speed, race earnings, and finish position. Principal component analysis was used to create orthogonal scores from multiple collinear variables for inclusion in the models. RESULTS: Furosemide was administered to 16,761 (74.2%) horses. Horses that received furosemide raced faster, earned more money, and were more likely to win or finish in the top 3 positions than horses that did not. The magnitude of the effect of furosemide on estimated 6-furlong race time varied with sex, with the greatest effect in males. When comparing horses of the same sex, horses receiving furosemide had an estimated 6-furlong race time that ranged from 0.56 +/- 0.04 seconds (least-squares mean +/- SE) to 1.09 +/- 0.07 seconds less than that for horses not receiving furosemide, a difference equivalent to 3 to 5.5 lengths. CONCLUSIONS AND CLINICAL RELEVANCE: Because of the pervasive use of furosemide and its apparent association with superior performance in Thoroughbred racehorses, further consideration of the use of furosemide and investigation of its effects in horses is warranted. PMID- 10476716 TI - Percutaneous lung biopsy in cattle. AB - OBJECTIVE: To determine safety of lung biopsy in healthy cattle and compare 3 biopsy instruments for suitability of use. DESIGN: Prospective study. ANIMALS: 60 healthy cows. PROCEDURE: Bilateral lung biopsy through the ninth intercostal space was attempted in all cows. The procedure was performed with cows standing and restrained by an assistant. To study the short-term effects of lung biopsy, 50 cows were slaughtered 1 day after the procedure and examined. To study the long-term effects of lung biopsy, the remaining 10 cows were examined daily for 10 days and then slaughtered. Three biopsy instruments--the Autovac biopsy cannula, the Tru-Cut needle, and the Bard Biopty-System--were evaluated. RESULTS: 2 cows collapsed momentarily during the procedure but recovered quickly. The remaining cows did not develop adverse effects, and general attitude and condition were not affected. Postmortem examination of the 50 cows slaughtered 1 day after the procedure revealed lesions < or = 5.0 mm long and with a surface area < or = 4.0 mm2 at 108 of 154 (71%) biopsy sites involving the pulmonary pleura. Biopsy specimens were obtained during 48 of 50 (96%) procedures in which the Tru-Cut needle was used, during 38 of 39 (97%) procedures in which the Bard Biopty-System was used, and during only 1 of 11 biopsy procedures in which the Autovac cannula was used. At postmortem examination of the 10 cows slaughtered 10 days after the procedure, lesions were seen as small scars; adhesions were not observed. CONCLUSIONS AND CLINICAL RELEVANCE: Percutaneous lung biopsy is a safe procedure in healthy cows. PMID- 10476717 TI - Feline interstitial cystitis. PMID- 10476718 TI - Sporotrichosis. PMID- 10476719 TI - Nosocomial bloodstream infections in United States hospitals: a three-year analysis. AB - Nosocomial bloodstream infections are important causes of morbidity and mortality. In this study, concurrent surveillance for nosocomial bloodstream infections at 49 hospitals over a 3-year period detected >10,000 infections. Gram positive organisms accounted for 64% of cases, gram-negative organisms accounted for 27%, and 8% were caused by fungi. The most common organisms were coagulase negative staphylococci (32%), Staphylococcus aureus (16%), and enterococci (11%). Enterobacter, Serratia, coagulase-negative staphylococci, and Candida were more likely to cause infections in patients in critical care units. In patients with neutropenia, viridans streptococci were significantly more common. Coagulase negative staphylococci were the most common pathogens on all clinical services except obstetrics, where Escherichia coli was most common. Methicillin resistance was detected in 29% of S. aureus isolates and 80% of coagulase-negative staphylococci. Vancomycin resistance in enterococci was species-dependent--3% of Enterococcus faecalis strains and 50% of Enterococcus faecium isolates displayed resistance. These data may allow clinicians to better target empirical therapy for hospital-acquired cases of bacteremia. PMID- 10476720 TI - Surveillance of antimicrobial use and antimicrobial resistance in United States hospitals: project ICARE phase 2. Project Intensive Care Antimicrobial Resistance Epidemiology (ICARE) hospitals. AB - The search for the means to understand and control the emergence and spread of antimicrobial resistance has become a public health priority. Project ICARE (Intensive Care Antimicrobial Resistance Epidemiology) has established laboratory based surveillance for antimicrobial resistance and antimicrobial use at a subset of hospitals participating in the National Nosocomial Infection Surveillance system. These data illustrate that for most antimicrobial-resistant organisms studied, rates of resistance were highest in the intensive care unit (ICU) areas and lowest in the outpatient areas. A notable exception was ciprofloxacin- or ofloxacin-resistant Pseudomonas aeruginosa, for which resistance rates were highest in the outpatient areas. For most of the antimicrobial agents associated with this resistance, the rate of use was highest in the ICU areas, in parallel to the pattern seen for resistance. These comparative data on use and resistance among similar areas (i.e., ICU or other inpatient areas) can be used as a benchmark by participating hospitals to focus their efforts at addressing antimicrobial resistance. PMID- 10476721 TI - National epidemiology of mycoses survey (NEMIS): variations in rates of bloodstream infections due to Candida species in seven surgical intensive care units and six neonatal intensive care units. AB - Candida species are the fourth most frequent cause of nosocomial bloodstream infections, and 25%-50% occur in critical care units. During an 18-month prospective study period, all patients admitted for > or = 72 hours to the surgical (SICUs) or neonatal intensive care units (NICUs) at each of the participant institutions were followed daily. Among 4,276 patients admitted to the seven SICUs in six centers, there were 42 nosocomial bloodstream infections due to Candida species (9.8/1,000 admissions; 0.99/1,000 patient-days). Of 2,847 babies admitted to the six NICUs, 35 acquired a nosocomial bloodstream infection due to Candida species (12.3/1,000 admissions; 0.64/1,000 patient-days). The following were the most commonly isolated Candida species causing bloodstream infections in the SICU: Candida albicans, 48%; Candida glabrata, 24%; Candida tropicalis, 19%; Candida parapsilosis, 7%; Candida species not otherwise specified, 2%. In the NICU the distribution was as follows: C. albicans, 63%; C. glabrata, 6%; C. parapsilosis, 29%; other, 3%. Of the patients, 30%-50% developed incidental stool colonization, 23% of SICU patients developed incidental urine colonization, and one-third of SICU health care workers' hands were positive for Candida species. PMID- 10476722 TI - Antimicrobial resistance in key bloodstream bacterial isolates: electronic surveillance with the Surveillance Network Database--USA. AB - To assess the prevalence of antimicrobial-resistant pathogens among the most common bloodstream isolates, we examined antimicrobial susceptibility data from The Surveillance Network Database-USA, an electronic surveillance system that collects data from 118 clinical microbiology laboratories across the United States. Between 1995 and 1997, resistance to both vancomycin and ampicillin was much more prevalent among Enterococcus faecium than Enterococcus faecalis, suggesting the need for laboratories to identify to species. When staphylococcal isolates were examined for reduced susceptibility to vancomycin (minimum inhibitory concentration = 4 microg/mL), the frequency was highest in methicillin resistant coagulase-negative staphylococci. We also learned that nonsusceptibility to ceftazidime in Klebsiella pneumoniae was more prevalent among isolates from blood (12.7%) than among isolates from urine (7.1%) or respiratory sources (9.3%). Although antimicrobial resistance is low overall for isolates of Escherichia coli from blood, the prevalence of cefoxitin resistance among ceftazidime-resistant strains (61.9%) suggests the action of mechanisms other than extended-spectrum beta-lactamase. PMID- 10476723 TI - Optimal use of modern antibiotics: emerging trends. AB - Development of new antimicrobial drugs is an essential component in the effort to remain ahead of emerging microbial resistance. However, when new antibiotics are used with unrestrained enthusiasm, a predictable consequence is the further expansion of resistance. This problem is well known to the infectious diseases specialist and is increasingly appreciated by the nonspecialist and the public. A far more sensible strategy is to identify new ways to use these drugs to increase the duration of their usefulness. New methods to optimize antibiotic selection, dose, and duration of therapy are being investigated, and application of some of these strategies has been shown to have a favorable impact on resistance. Much of the classic thinking of how to use antibiotics is changing, and these newer strategies may result in prolongation of the era of the "antibiotic miracle." PMID- 10476724 TI - Photo quiz. Diagnosis: tularemia. PMID- 10476725 TI - Baker's cyst infection: case report and review. PMID- 10476726 TI - Editorial response: Why a duck?--Or for that matter, why a cyst? PMID- 10476727 TI - Reduction in functional antibody activity against Streptococcus pneumoniae in vaccinated elderly individuals highly correlates with decreased IgG antibody avidity. AB - The pneumococcal polysaccharide vaccine is recommended as a means of preventing invasive disease in the elderly. We compared responses to the 23-valent polysaccharide vaccine in 46 previously unvaccinated, healthy, institutionalized elderly persons (mean age, 85.5 years) with those in 12 healthy younger adults (mean age, 37 years) by measuring prevaccination and postvaccination serum IgG antibody concentrations (by ELISA), functional antibody activity (by opsonophagocytosis), IgG antibody avidity, and passive protection in mice. Postvaccination IgG antibody concentrations for two serotypes (6B and 19F) of the five studied (4, 6B, 14, 19F, and 23F) were significantly lower in elderly than in younger adults; however, opsonophagocytic activity was significantly reduced for all serotypes in the elderly. Sera with reduced opsonophagocytic activity (titer, <64) correlated with low IgG antibody avidity and protected mice poorly against pneumococcal challenge. In elderly persons receiving polysaccharide vaccination, there was a significant reduction in the functionality of postvaccination antibodies, and this appeared to increase with advanced age. PMID- 10476728 TI - Editorial response: Predicting protection against encapsulated pathogens. PMID- 10476729 TI - Prosthetic joint infection: when can prosthesis salvage be considered? AB - Management of prosthetic joint infection (PJI) remains a therapeutic challenge. We retrospectively studied 69 infected total hip or knee arthroplasties managed between 1980 and 1996 in our institution. Treatment failure, defined as relapse of PJI in the first year following the last antimicrobial treatment, occurred for 14 patients (20.3%). None of the potentially contributive parameters analyzed was significantly predictive of treatment failure. Of the subgroup of 34 patients with PJI who initially underwent debridement with retention of the prosthesis, the 13 (38.2%) who did not require further surgical treatment had symptoms for a significantly shorter duration before debridement (4.85 vs. 54.24 days; P < .0001). Because debridement with retention of the prosthesis rarely enables control of PJI, this therapeutic approach should be considered only when the duration of symptoms is very short. PMID- 10476730 TI - Editorial response: Orthopedic prosthesis salvage. PMID- 10476731 TI - Escherichia coli O157:H7 outbreak associated with an improperly chlorinated swimming pool. AB - A cluster of gastrointestinal illnesses, including one case of hemolytic-uremic syndrome and one culture-confirmed Escherichia coli O157:H7 infection, followed a trailer park pool party. We interviewed a cohort of party attendees and park residents. A primary case was defined as the first gastrointestinal illness within a household between 5 July and 20 July in which the titer of IgG antibody to E. coli O157 (if determined) was elevated. Of 51 party attendees and trailer park residents, 18 developed a gastrointestinal illness, including 10 who met the definition of a primary case. Swimming in the pool significantly increased the risk of primary illness (relative risk = 6.3; 95% confidence interval = 1.8 18.9). No other exposure was significantly associated with primary illness, after pool exposure was controlled for. The implicated pool had little to no chlorine added during the period of 4-10 July. This outbreak provides new evidence of the importance of proper pool maintenance in controlling the spread of E. coli O157:H7. PMID- 10476733 TI - Editorial response: Treatment of cystic echinococcosis--improving but still limited. PMID- 10476732 TI - Long-term evaluation of patients with hydatidosis treated with benzimidazole carbamates. AB - Four hundred forty-eight patients with 929 Echinococcus granulosus hydatid cysts received 3- to 6-month continuous cycles of mebendazole or albendazole treatment and underwent prolonged follow-up by clinical visits and imaging studies (range, 1-14 years) to assess the long-term outcome of treatment. Degenerative changes and relapse were assessed by imaging techniques. At the end of therapy, 74.1% of the hydatid cysts showed degenerative changes. These were more frequent in albendazole-treated than in mebendazole-treated cysts (82.2% vs. 56.1%; P < .001). During long-term follow-up, 104 cysts (22%) had degenerative changes that progressed, whereas 163 cysts (approximately 25%) relapsed. The percentages of relapses in the two drug-treated groups were almost the same. Relapses occurred more frequently in type II cysts of the liver. Cysts recurred most often (78.5%; P < .001) within the first 2 years after treatment ended. Further chemotherapy cycles induced degenerative changes in >90% of relapsed cysts without inducing more frequent or more severe side effects than those observed during the initial cycles. PMID- 10476734 TI - Antibiotic prescribing by pediatricians for respiratory tract infection in children. AB - To examine antimicrobial prescribing rates for viral respiratory tract infections by primary care pediatricians in the greater Toronto area, charts were reviewed for the week of 17-21 February 1997 at 61 pediatricians' offices. Antibiotics were considered appropriate if the diagnosis was compatible with bacterial infection. A total of 3,585 patient visits were reviewed. The common cold was the most common respiratory tract syndrome leading to an office visit (1,317 visits). The overall rate of appropriate antibiotic prescribing was 89.5%. There was no significant difference in prescribing when physicians were compared by year of graduation from medical school, sex, or location of training. Diagnostic codes (ICD-9 [International Classification of Diseases, 9th edition] codes) did not match the chart diagnosis in 41% of cases. Toronto primary care pediatricians appear to have a lower rate of inappropriate antibiotic prescribing than do primary care physicians in other regions of Canada and the United States. PMID- 10476735 TI - Editorial response: Judicious use of antimicrobial agents for respiratory tract infections in children--why are the pediatricians in Toronto doing so well? PMID- 10476736 TI - Penicillin resistance and other predictors of mortality in pneumococcal bacteremia in a population with high human immunodeficiency virus seroprevalence. AB - Rates of invasive disease caused by penicillin-resistant pneumococci are rising. Previous reports have found no association between resistant pneumococci and increased mortality. To evaluate the impact of penicillin resistance and other variables on mortality, we retrospectively studied all cases of pneumococcal bacteremia identified by our microbiology laboratory from 1 January 1992 through 31 December 1996. There were 462 cases of pneumococcal bacteremia in 432 patients. The mean age was 35 years; 55% of the cases occurred in male patients, 58% were in black patients, and 40% were in Hispanic patients. One-half of the cases occurred in patients with documented human immunodeficiency virus (HIV) infection. Penicillin resistance was first noted in 1994 and increased yearly, accounting for 17% of 1996 isolates. Of all resistant isolates, 65% were resistant to penicillin at a high level. The overall mortality was 17%. On multivariate analysis, high-level penicillin resistance, older age, severe disease, multilobar infiltrates and/or effusion(s) on chest roentgenogram, and Hispanic ethnicity were independent predictors of mortality in pneumococcal bacteremia. In HIV-infected patients, a CD4 cell count below the median just missed statistical significance. This is the first report demonstrating penicillin resistance as an independent predictor of mortality among patients with pneumococcal bacteremia. PMID- 10476737 TI - Vitamin A deficiency and other nutritional indices during pregnancy in human immunodeficiency virus infection: prevalence, clinical correlates, and outcome. Women and Infants Transmission Study Group. AB - Vitamin A levels in plasma and other nutritional indices were measured during pregnancy for 449 women enrolled in a multicenter cohort study of mother-to infant transmission of human immunodeficiency virus type 1 (HIV-1). During the third trimester, 29.6% of the women had low (20 to <30 microg/dL) and 11.1% had very low (<20 microg/dL) vitamin A levels. Vitamin A and body mass index, serum albumin levels, and hemoglobin levels were weakly correlated. After adjustment for other covariates, women with low and very low vitamin A levels before the third trimester were more likely to deliver infants with low birth weight (<2500 g) than were those with higher levels (odds ratio [OR], 4.58; 95% confidence interval [CI], 1.57-13.4; and OR, 6.99; 95% CI, 1.09-45.0, respectively). However, there was no statistically significant association between vitamin A level and mother-to-infant transmission of HIV-1. Anemia and low body mass index before the third trimester were associated with an increased risk of transmission in univariate analyses but not in multivariate analyses. PMID- 10476738 TI - Enteroaggregative Escherichia coli as a cause of traveler's diarrhea: clinical response to ciprofloxacin. AB - The purpose of this study was to determine the role of enteroaggregative Escherichia coli (EAEC) in the development of traveler's diarrhea and the clinical response of patients with EAEC diarrhea following treatment with ciprofloxacin. Sixty-four travelers with diarrhea and no other recognized enteropathogen were enrolled in treatment studies in Jamaica and Mexico from July 1997 to July 1998. EAEC was isolated from 29 travelers (45.3%). There was a significant reduction in the duration of posttreatment diarrhea in the 16 patients treated with ciprofloxacin, as compared with that in the 13 patients who received placebo (mean of 35.3 versus 55.5 hours; P = .049). There was a nonsignificant reduction in the mean number of unformed stools passed during the 72 hours after enrollment in the ciprofloxacin-treated group (5.6), as compared with that in the placebo group (7.5) (P = .128). This study provides additional evidence that EAEC should be considered as a cause of antibiotic-responsive traveler's diarrhea. PMID- 10476739 TI - Postoperative spondylodiskitis: etiology, clinical findings, prognosis, and comparison with nonoperative pyogenic spondylodiskitis. AB - We studied 31 cases of postoperative pyogenic spondylodiskitis (POS), comparing them with 72 cases of nonpostoperative pyogenic spondylodiskitis (NPOS). POS represents 30.1% of cases of pyogenic spondylodiskitis. The onset of symptoms occurred an average (+/-SD) of 27.7 (+/- 25.3) days following surgery. Predisposing factors were less frequent in POS than NPOS cases (P = .002). Neurological complications and inflammatory signs in the spine were more frequent with POS than with NPOS (P = .002 and P < .00001). Coagulase-negative Staphylococcus and anaerobic bacteria were more frequent in POS than in NPOS (P = .0001 and P = .05). Percutaneous bone biopsies yielded the etiology in 66.7% of cases, open bone biopsies in 100%, blood cultures in 55.6%, and cultures of adjacent foci in 94.4%. Eleven patients (35.5%) were cured with antimicrobial treatment, but surgical treatment was necessary in 64.5%. No relapses or deaths were recorded. Seventeen patients (54.8%) had severe functional sequelae, which were associated with inflammatory signs in the spine (P = .033), higher levels of leukocytosis (P = .05), higher erythrocyte sedimentation rates (P = .05), and paravertebral abscesses (P = .04). PMID- 10476740 TI - Morbidity associated with long-term use of totally implantable ports in patients with AIDS. AB - To determine the morbidity associated with long-term use of a totally implantable central venous access device (Port-A-Cath [PAC]) in patients with AIDS, we studied 68 consecutive patients with AIDS requiring 79 such devices for long-term use, inserted over a period of 5 years. The total number of PAC-days was 20,159. At least one PAC-related complication occurred with 40 of 79 PACs (50.6% [95% confidence interval (CI): 39.6%-61.6%]), and 16 devices (20.2% [95% CI, 11.4% 29.0%]) had to be removed because of complications. Device-related infection occurred with 33 of 79 PACs (41.7% [95 CI, 30.8%-52.6%]). The predominant infection occurring with PACs was chamber infection, with an incidence of 0.16 per 100 PAC-days. The predominant organisms isolated from patients with chamber infections but also from those with device-related bacteremia were gram-positive cocci (79.4%). The presence of neutropenia (odds ratio [OR] = 9.72; 95% CI, 3.0 31.3; P < .001) and a CD4 cell count lower than 0.025 x 10(9)/L (OR = 6.14; 95% CI, 1.9-19.2; P = .002) were independent predictors of infection. The antibiotic lock technique was associated with decreased device loss when compared with isolated systemic antibiotic therapy (OR = 0.05; 95% CI, 0.0-0.59; P = .008). This technique may be useful to treat PAC infection in patients with AIDS, for whom the risk of PAC-related complications is very high. PMID- 10476741 TI - Clinical characteristics and molecular epidemiology associated with imipenem resistant Klebsiella pneumoniae. AB - Eight patients were infected or colonized with imipenem-resistant Klebsiella pneumoniae (IRKP) from December 1994 to November 1995. Initial Klebsiella isolates were susceptible to imipenem but resistant to all cephalosporins, aminoglycosides, and beta-lactam inhibitor combinations. All patients had been in the surgical intensive care unit and had undergone abdominal surgery or tracheostomy during hospitalization. The average age of the patients was 71 years (range, 41-81 years). All patients were treated with imipenem for 5 to 36 days, and IRKP was recovered from each during or after therapy. Pulsed-field gel electrophoresis (PFGE) of the IRKP isolates revealed three distinct clonal patterns. Paired sequential isolates of imipenem-susceptible K. pneumoniae and IRKP from two patients had identical PFGE patterns, suggesting the development of clonal stepwise resistance to imipenem during therapy. Thus, imipenem resistance in Klebsiella may occur when this agent is used for treatment of infection due to ceftazidine- and aminoglycoside-resistant strains. PMID- 10476742 TI - Comparative value of colonic biopsy and intraluminal fluid culture for diagnosis of bacterial acute colitis in immunocompetent patients. Infectious Colitis Study Group. AB - We compared the yield of intraluminal fluid culture to that of biopsy specimens obtained during colonoscopy for the diagnosis of bacterial colitis in 93 immunocompetent patients with a recent episode of diarrhea and macroscopic lesions of colitis. Stool culture findings were also available for 68 patients. At least one bacterial pathogen was isolated from the biopsy specimen, intraluminal fluid, or stool from 48 patients (51.6%). Salmonella species, Clostridium difficile, Klebsiella oxytoca, Shigella species, and Campylobacter species were recovered from 16 (17.2%), 15 (16.1%), 8 (8.6%), 7 (7.5%), and 4 (4.3%) of the patients, respectively. One Shigella species and one K. oxytoca strain were isolated from biopsy specimens but not from intraluminal fluid, and intraluminal fluid was the only positive specimen in 12 cases (yielding 1 Salmonella species, 2 Shigella species, 2 K. oxytoca, and 7 C. difficile isolates). In nine cases out of 10, toxin B was detected only in intraluminal fluid. A correlation of 91.2% was observed between stool and intraluminal fluid cultures for Salmonella, Shigella, and Campylobacter species isolations. Culture of biopsy specimens adds little to the diagnosis of infectious colitis, and stools and intraluminal fluids appear to have comparable value. PMID- 10476743 TI - Lack of efficacy of oral bacitracin plus doxycycline for the eradication of stool colonization with vancomycin-resistant Enterococcus faecium. AB - In a prospective observational cohort study designed to assess the role of oral bacitracin solution plus doxycycline in the eradication of intestinal carriage of vancomycin-resistant Enterococcus faecium (VREF) in patients on a renal ward, rectal swab specimens were obtained from 15 treated and 24 control patients. Cultures of the rectal swabs were negative for 15 (100%) of the antibiotic treated vs. eight (33.3%) of the untreated patients (P < .001) on day 14. However, follow-up for a mean of 127 and 130 days revealed 9 of 15 (60%) and 15 of 24 (62.5%) in the treated and untreated cohorts (P = .86), respectively, carried VREF intermittently or persistently. Quantitative VREF stool cultures in the treated cohort revealed an initial 3.1-log10/g decrease, but there was an increase to pretreatment levels at 2-4 and 5-7 weeks post-treatment (7.8 and 7.4 log10/g). Oral bacitracin and doxycycline were not efficacious in reducing the carriage of VREF beyond the 2-week interval during which they were given. PMID- 10476744 TI - Clostridium difficile colitis associated with infant botulism: near-fatal case analogous to Hirschsprung's enterocolitis. AB - We present the first five reported cases of Clostridium difficile-associated diarrhea (CDAD) in children with infant botulism caused by Clostridium botulinum. We compare two fulminant cases of colitis in children with colonic stasis, the first caused by infant botulism and the second caused by Hirschsprung's disease. In both children, colitis was accompanied by hypovolemia, hypotension, profuse ascites, pulmonary effusion, restrictive pulmonary disease, and femoral-caval thrombosis. Laboratory findings included pronounced leukocytosis, hypoalbuminemia, hyponatremia, coagulopathy, and, when examined in the child with infant botulism, detection of C. difficile toxin in ascites. CDAD recurred in both children, even though difficile cytotoxin was undetectable in stool after prolonged initial therapy. Four children who had both infant botulism and milder CDAD also are described. Colonic stasis, whether acquired, as in infant botulism, or congenital, as in Hirschsprung's disease, may contribute to the susceptibility to and the severity of CDAD. PMID- 10476745 TI - A comparison of chloramphenicol, trimethoprim-sulfamethoxazole, and doxycycline with doxycycline alone as maintenance therapy for melioidosis. AB - A prospective, open, randomized, comparative treatment trial was conducted to compare the therapeutic efficacy of the conventional four-drug combination (chloramphenicol, trimethoprim-sulfamethoxazole, and doxycycline) with that of doxycycline alone in oral maintenance treatment of melioidosis. Adult Thai patients with culture-confirmed melioidosis were randomized to receive treatment with either regimen for a minimum of 12 weeks, usually following intravenous treatment of severe disease. The main outcome measure was culture-confirmed relapse. One hundred sixteen patients were enrolled; 109 had culture-confirmed melioidosis, and 87 were considered evaluable (43 had received doxycycline). Culture-confirmed relapse occurred in one patient randomized to the conventional regimen and in 11 (25.6%) randomized to the doxycycline regimen (P = .009), and treatment failed for 8 (18.2%) versus 20 (46.5%), respectively (P = .009). Adverse effects occurred in 26% of patients overall. Doxycycline alone cannot be recommended for a first-line regimen of oral maintenance treatment of melioidosis. PMID- 10476746 TI - Comparison of imipenem and ceftazidime as therapy for severe melioidosis. AB - An open, prospective, randomized, comparative treatment trial was conducted to compare the therapeutic efficacy of high-dose intravenous imipenem and ceftazidime for acute severe melioidosis. Adult Thai patients with suspected acute, severe melioidosis were randomized to receive either imipenem, at a dosage of 50 mg/(kg x d), or ceftazidime, at a dosage of 120 mg/(kg x d), for a minimum of 10 days. The main outcome measures were death or treatment failure. Of the 296 patients enrolled, 214 had culture-confirmed melioidosis, and 132 (61.7%) of them had positive blood cultures. Mortality among patients with melioidosis was 36.9% overall. There were no differences in survival overall (P = .96) or after 48 hours (P = .3). Treatment failure after 48 hours was more common among patients treated with ceftazidime (P = .011). Both treatments were well tolerated. Imipenem is a safe and effective treatment for acute severe melioidosis and may be considered an alternative to ceftazidime. PMID- 10476748 TI - Coxiella burnetii pericarditis: report of 15 cases and review. AB - Q fever is characterized by its clinical polymorphism, and pericarditis associated with Q fever has occasionally been described. Herein we report 15 cases of Coxiella burnetii pericarditis, 9 from our data bank and 6 encountered within the past 12 months. Three patients presented with life-threatening tamponade. We compare our cases with the 18 previously reported and with 60 Q fever-matched controls at our center. This study showed that Q fever pericarditis can present as acute as well as chronic disease; we describe relapse after 6 months in association with a serological profile compatible with the chronic form of disease (phase I C. burnetii IgG titer of > or = 800). Discriminant factors among patients and controls are age of > 52 years (adjusted odds ratio [OR], 5.66), the occurrence of general symptoms such as arthralgias or myalgias (adjusted OR, 6.54), and a normal erythrocyte sedimentation rate (adjusted OR, 16.37). No specific symptoms or underlying cardiac predispositions are observed. PMID- 10476747 TI - Human granulocytic ehrlichiosis in Northern California: two case descriptions with genetic analysis of the Ehrlichiae. AB - We report two cases of human granulocytic ehrlichiosis (HGE) that occurred in northern California in summer 1998. Patients had fever, malaise, and myalgia, reported tick bites, had moderate thrombocytopenia, and had normal or slightly elevated liver enzyme activities. Ehrlichial inclusions were observed in the blood of one patient, and HGE-agent DNA was amplified by PCR from both patients. Genetically, the strains resembled horse isolates from northern California. The close spatial and temporal proximity of the two new cases may be due to a nidus of infection in the area or heightened surveillance by local physicians. PMID- 10476749 TI - Prediction of microbial infection and mortality in medical patients with fever: plasma procalcitonin, neutrophilic elastase-alpha1-antitrypsin, and lactoferrin compared with clinical variables. AB - Fever suggests the likelihood of severe microbial infection. Abnormal temperature, tachycardia, tachypnea, and abnormal white blood cell counts define the systemic inflammatory response syndrome (SIRS). In 300 hospitalized medical patients with fever, we determined clinical variables and procalcitonin, elastase alpha1-antitrypsin, and lactoferrin levels in plasma. Of the patients, 71% had clinical infection (by clinical judgment) and 44% had microbial infection (by microbiological testing). SIRS occurred in 95%, and the 28-day mortality rate was 9%. The sensitivity for predicting microbial infection, bacteremia, and mortality was less but the specificity was greater for supranormal procalcitonin, elastase alpha1-antitrypsin, and lactoferrin levels than for SIRS. The area under the receiver operating characteristic curve (AUC) for microbial infection was higher for procalcitonin and elastase-alpha1-antitrypsin levels than for clinical variables and lactoferrin level. The AUC for bacteremia was also higher for inflammatory factors (>0.70; P < .001) than for clinical variables. The AUC for mortality (P < .05) was 0.79 for the respiratory rate, 0.69 for elastase-alpha1 antitrypsin level, 0.65 for heart rate, 0.61 for procalcitonin level, and 0.60 for white blood cell count. In febrile medical patients, plasma procalcitonin and elastase-alpha1-antitrypsin levels may predict microbial infection and bacteremia better than (and mortality as well as) do clinical symptoms. PMID- 10476750 TI - Risk factors for melioidosis and bacteremic melioidosis. AB - A case-control study was conducted in four hospitals in northeastern Thailand to identify risk factors for melioidosis and bacteremic melioidosis. Cases were patients with culture-proven melioidosis, and there were two types of controls (those with infections, i.e., with community-acquired septicemia caused by other bacteria, and those without infection, i.e., randomly selected patients admitted with noninfectious diseases to the same hospitals). Demographic data, clinical presentations, and suspected risk factors were analyzed. Diabetes mellitus, preexisting renal diseases, thalassemia, and occupational exposure, classified by the soil and water risk assessment, were confirmed to be significant risk factors for melioidosis and bacteremic melioidosis. Only diabetes mellitus was a significant factor associated with bacteremic melioidosis, as compared with nonbacteremia. A significant interaction was found between diabetes mellitus and occupational exposure. Thus, diabetic rice farmers would be the most appropriate population group for targeted control measures such as vaccination in the future. PMID- 10476751 TI - Assessment of measuring circulating levels of interleukin-6, interleukin-8, C reactive protein, soluble Fc gamma receptor type III, and mannose-binding protein in febrile children with cancer and neutropenia. AB - Circulating levels of interleukin (IL)-6, IL-8, soluble Fc gamma receptor type III (sFc gammaRIII), mannose-binding protein (MBP), and C-reactive protein (CrP) were assessed among febrile children with cancer and neutropenia. Levels of IL-6, IL-8, sFc gammaRIII, MBP, and CrP were measured in serum from 56 pediatric cancer patients at the time of admission for 121 episodes of febrile neutropenia (88 febrile episodes without identifiable source, 5 clinically documented infections, 20 episodes of bacteremia due to gram-positive and 5 due to gram-negative organisms, and 3 fungal infections). IL-6 and IL-8 levels were higher in patients with either bacteremia due to gram-negative organisms or fungal infections than in patients with febrile episodes without an identifiable source (P < .00001 for each). IL-6 and IL-8 levels were higher in children with bacteremia due to gram negative organisms than in those with bacteremia due to gram-positive organisms (P = .0011 and P = .0003, respectively). The measured levels of CrP, MBP, and sFc gammaRIII were not useful for identifying the type of infection. These preliminary results show the potential usefulness of IL-6 and IL-8 as early indicators for life-threatening infections in febrile cancer patients with neutropenia. PMID- 10476752 TI - Influence of environmental temperature on incidence of indinavir-related nephrolithiasis. AB - We analyzed the influence of temperature, humidity, and atmospheric pressure on the 1-year incidence of nephrolithiasis among human immunodeficiency virus type 1 infected patients treated with indinavir. One hundred three patients (13.6%) developed 326 episodes of nephrolithiasis. Eighty-two patients (79.6%) had more than one episode (range, two to seven episodes). The overall incidence ranged from 0 to 10.2 episodes per 100 patients exposed per month. There was a significant correlation between temperature and the overall incidence of nephrolithiasis and the incidence of recurrences but not with the incidence of first episodes. Nephrolithiasis was not related to humidity or atmospheric pressure. Our data support the standard recommendation of drinking at least 1.5 L of water daily to prevent nephrolithiasis in most patients treated with indinavir irrespective of meteorologic factors. However, the risk of nephrolithiasis is higher for a certain subgroup of patients when the environment is hot irrespective of adequate water intake. PMID- 10476753 TI - Clinical characteristics of Chlamydia pneumoniae infection as the sole cause of community-acquired pneumonia. AB - The clinical characteristics of 26 patients with community-acquired pneumonia due to Chlamydia pneumoniae as the only identified pathogen who required hospitalization were evaluated. Most patients (18) had reinfection based on serological results. The mean age of the patients was 55 years (38 years, patients with primary infection; 63 years, patients with reinfection), and the gender representation was equal. Generally, illness was mild and associated with limited temperature elevation and nonspecific symptoms. The presence of comorbid illnesses and the requirement of supplemental oxygen therapy were the most common criteria for hospital admission. PMID- 10476754 TI - Pulmonary toxoplasmosis in bone marrow transplant recipients: report of two cases and review. AB - Toxoplasma gondii may cause disseminated disease in bone marrow transplant (BMT) recipients. Pulmonary toxoplasmosis in BMT patients is rarely described. Mortality rates of >90% have been previously reported. Since pulmonary toxoplasmosis is extremely difficult to diagnose, it is very often detected only at autopsy. Two cases of pulmonary toxoplasmosis in BMT recipients that were diagnosed by visualization of T. gondii tachyzoites in bronchoalveolar lavage fluid and by a new semi-nested PCR method amplifying 18S rRNA from bronchoalveolar lavage fluid are presented, and the literature on pulmonary toxoplasmosis in BMT patients is reviewed. PMID- 10476755 TI - Survey of three bacterial louse-associated diseases among rural Andean communities in Peru: prevalence of epidemic typhus, trench fever, and relapsing fever. AB - Typhus and other louse-transmitted bacterial infections in Peruvian sierra communities are known to occur but have not recently been assessed. In this study, 194 of 1,280 inhabitants of four villages in Calca Province in the Urubamba Valley were included. Thirty-nine (20%) of the 194 volunteers had antibodies to Rickettsia prowazekii, whereas 24 (12%) had antibodies to Bartonella quintana and 2 against Borrelia recurrentis. There was a significant correlation between the presence of infesting ectoparasites and antibodies to R. prowazekii, as well as between antibodies to R. prowazekii and ectoparasite infestation and fever in the previous 6 months. The proportion of inhabitants infested with ectoparasites was significantly higher in the highest-altitude village than in the other three villages. Two volunteers' antibody levels suggested a recent typhus infection, but only B. quintana DNA was amplified from lice. Epidemic typhus remains extant in the area, and B. quintana infections were encountered and documented for the first time in South America. PMID- 10476756 TI - Respiratory viral antigens in autopsy lung tissue specimens from patients with cancer or myocardial infarction. AB - Using immunoenzyme histochemical analysis, we retrospectively examined lung tissue specimens obtained at autopsy from 118 patients with cancer who had received chemotherapy and 20 patients who had died after myocardial infarction. Respiratory viral antigens were demonstrated in lung tissue specimens from eight of 118 cancer patients and two of 20 myocardial infarction patients. Most of the patients with demonstrable viral antigens were febrile and had signs of pulmonary infection, but in no case was pulmonary viral infection considered clinically. The following viral antigens were demonstrated: influenza A virus (6 patients), respiratory syncytial virus (2), influenza B virus (1), and parainfluenza virus type 1 (1). PMID- 10476757 TI - Alterations in serum levels of lipids and lipoproteins with indinavir therapy for human immunodeficiency virus-infected patients. AB - Alterations in lipid metabolism have been associated with the use of protease inhibitors. Sequential lipid analyses were performed on serum samples from human immunodeficiency virus-infected antiretroviral-naive patients who received indinavir in combination with two nucleoside reverse transcriptase inhibitors. Serum levels of cholesterol, triglycerides, high-density lipoproteins (HDLs), and low-density lipoproteins (LDLs) were measured at baseline and at periodic intervals. After 48 weeks of indinavir therapy, mean serum levels +/- SD rose as follows: cholesterol, from 167.2 +/- 36.0 to 206.3 +/- 32.4 mg/dL (P < .0005); triglycerides, from 110.4 +/- 47.5 to 158.4 +/- 72.5 mg/dL (P < .0101); and LDLs, from 106.6 +/- 35.1 to 136.1 +/- 31.6 mg/dL (P = .0029). There was no significant change in the serum HDL fraction. Mean serum lipoprotein (a) levels +/- SD rose from 6.5 +/- 1.4 to 9.6 +/- 2.0 mg/dL after 30 weeks (P = .0695). Potential mechanisms for the noted increases include alterations in serum lipoprotein lipase activity or changes in hepatic lipid metabolism. The clinical significance of these changes remains to be determined. PMID- 10476758 TI - Therapeutic options for malacoplakia secondary to Escherichia coli infection. PMID- 10476759 TI - A "query" pancreatitis in a young shepherdess: an uncommon manifestation of acute Q fever. PMID- 10476760 TI - Prolonged fever caused by parvovirus B19-induced meningitis: case report and review. PMID- 10476761 TI - Cardiac cirrhosis with cellulitis caused by Burkholderia cepacia bacteremia. PMID- 10476762 TI - Escherichia coli meningitis in a human immunodeficiency virus-infected man after outpatient hemorrhoidectomy. PMID- 10476763 TI - Symptomatic junctional bradycardia after treatment with nelfinavir. PMID- 10476764 TI - Disseminated infection after bacille Calmette-Guerin instillation for treatment of bladder carcinoma. PMID- 10476765 TI - Chronic Chlamydia pneumoniae infection: a treatable cause of chronic fatigue syndrome. PMID- 10476766 TI - Acute fever and petechial rash associated with influenza A virus infection. PMID- 10476767 TI - Ecthyma secondary to herpes simplex virus infection. PMID- 10476768 TI - Severe hepatic failure related to nevirapine treatment. PMID- 10476769 TI - Antigenic drift of influenza A (H3N2) virus in a persistently infected immunocompromised host is similar to that occurring in the community. PMID- 10476770 TI - Takayasu's arteritis in a human immunodeficiency virus-infected adolescent. PMID- 10476771 TI - Stavudine-induced macrocytosis during therapy for human immunodeficiency virus infection. PMID- 10476772 TI - Refractory craniofacial actinomycetoma due to Streptomyces somaliensis that required salvage therapy with amikacin and imipenem. PMID- 10476774 TI - Pseudomonas aeruginosa--still a frequent pathogen in patients with cancer: 11 year experience at a comprehensive cancer center. PMID- 10476773 TI - Failure of treatment of pneumonia associated with highly resistant pneumococci in a child. PMID- 10476775 TI - Central nervous system disturbances following clarithromycin ingestion. PMID- 10476776 TI - Topical paromomycin/methylbenzethonium chloride plus parenteral meglumine antimonate as treatment of American cutaneous leishmaniasis: controlled study. PMID- 10476777 TI - Congenital tuberculosis in a neonatal intensive care. PMID- 10476778 TI - Incidence and spectrum of AIDS-defining illnesses among persons treated with antiretroviral drugs. PMID- 10476779 TI - Ultrastructural changes of photoreceptor synaptic ribbons in relation to time of day and illumination. AB - PURPOSE: Electron microscopic sections through rod and cone ribbon synapses reveal mainly rodlike synaptic ribbon profiles, but a few unusual spherical and club-shaped profiles also occur. To elucidate the meaning of the latter two forms, the authors have investigated these ribbon synapses at different times during the 24-hour cycle and under various lighting conditions. METHODS: The various types of ribbon profiles were counted, and their sizes were measured by means of transmission electron microscopy in retinas of male BALB/c mice exposed to 12 hours light (lights on at 6 AM) and 12 hours dark (LD 12:12), continuous light, or continuous darkness for 4 days. RESULTS: A 24-hour study of mice exposed to LD 12:12 showed that spherical and club-shaped profile numbers ranged from 0% to 29%, depending on the time of day. They reached a maximum at 3 hours after light onset, followed by a gradual decrease to approach zero at night and reappearing after light onset the next morning. After 4 days of continuous light, the spherical profiles were significantly decreased in number (examined at 9 AM). After continuous darkness, the spherical and club-shaped profiles were significantly reduced in number. Administration of 4 hours of light after 92 hours of continuous darkness restored the number of spherical and club-shaped profiles to normal values. The rodlike ribbon profiles were found to be longer in darkness than in light. In rod terminals containing spherical profiles, the rodlike ribbon profiles were shorter than in terminals without spherical profiles. CONCLUSIONS. The club-shaped and the spherical profiles were related to the turnover of the synaptic ribbons. Soon after light exposure in the morning, the synaptic ribbons formed distal swellings, giving rise to club-shaped profiles and a decrease in length. The swellings appeared to bud off, thus forming spherical synaptic bodies. This article discusses whether these changes are signs of degradation of spent ribbons, or whether they play a physiological role related to the inactivation of the ribbon synapses after light exposure. PMID- 10476780 TI - Effects of hyaluronan lyase, hyaluronidase, and chondroitin ABC lyase on mammalian vitreous gel. AB - PURPOSE: To determine the effects of enzymes on mammalian vitreous gel and to thus infer the structural roles of hyaluronan and chondroitin sulfate in the gel. METHODS: The wet weights of bovine vitreous gels were compared before and after incubation with Streptomyces hyaluronan lyase, chondroitin ABC lyase, testicular hyaluronidase, or buffer alone. The extent of hyaluronan depolymerization was determined by chromatography and that of chondroitin sulfate depolymerization by western blot analysis. RESULTS: After digestion with Streptomyces hyaluronan lyase (30 U/gel), the gel wet weight was the same as that of controls (incubated with buffer alone) despite 94% of the hyaluronan having been depolymerized; when digested with 100 U/gel, the gel wet weight decreased (to 57% of original wet weight versus 86% for controls, P = < 0.001) and hyaluronan was completely depolymerized. Chondroitin ABC lyase digestion (0.2 U/gel) resulted in a slight reduction in gel wet weight (90% versus 96%, P = < 0.001) and depolymerization of 88% of the hyaluronan; the presence of fully digested chondroitin sulfate chains was established. Digestions with 100 and 500 U/gel of testicular hyaluronidase resulted in a decrease (P = < 0.001, both cases) in gel wet weight (53% versus 82%, 100 U/gel; 57%, versus 86%, 500 U/gel) with 75% and 97% hyaluronan depolymerization, respectively. CONCLUSIONS: Depolymerization of all vitreous hyaluronan and of chondroitin sulfate resulted in gel wet weight reduction but not gel destruction. Digestion with 30 U/gel of Streptomyces hyaluronan lyase revealed a small pool (6%) of relatively enzyme-resistant hyaluronan that specifically contributed toward maintaining gel wet weight. PMID- 10476781 TI - Refractive errors in a black adult population: the Barbados Eye Study. AB - PURPOSE: To describe the prevalence of refractive errors in a black adult population. METHODS: The Barbados Eye Study, a population-based study, included 4709 Barbados-born citizens, or 84% of a random sample, 40 to 84 years of age. Myopia and hyperopia were defined as a spherical equivalent <-0.5 diopters and >+0.5 diopters, respectively, based on automated refraction. Analyses included 4036 black participants without history of cataract surgery. Associations with myopia and hyperopia were evaluated in logistic regression analyses. RESULTS: The prevalence of myopia was 21.9% and was higher in men (25.0%) than in women (19.5%). The prevalence of hyperopia was 46.9% and was higher in women (51.8%) than in men (40.5%). The prevalence of myopia decreased from 17% in persons 40 to 49 years of age to 11% in those 50 to 59 years of age, but increased after 60 years of age. The prevalence of hyperopia increased from 29% at 40 to 49 years of age to 65% at 50 to 59 years of age, and tended to decline thereafter. A higher prevalence of myopia was positively associated (P < 0.05) with lifetime occupations requiring nearwork, nuclear opacities, posterior subcapsular opacities, glaucoma, and ocular hypertension. Factors associated with hyperopia were the same as for myopia, except for occupation, and in the opposite direction. CONCLUSIONS: High prevalences of myopia and hyperopia were found in this large black adult population. The prevalence of myopia (hyperopia) increased (decreased) after 60 years of age, which is inconsistent with data from other studies. The high prevalence of age-related cataract, glaucoma, and other eye conditions in the Barbados Eye Study population may contribute to the findings. PMID- 10476782 TI - Lacrimal gland HGF, KGF, and EGF mRNA levels increase after corneal epithelial wounding. AB - PURPOSE: To evaluate the effect of corneal epithelial wounding on lacrimal gland expression of hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), and epidermal growth factor (EGF) in the rabbit model. METHODS: Rabbits had corneal epithelial scrape injuries, and the lacrimal gland was removed at different times after wounding. HGF, KGF, and EGF mRNA expression was examined by quantitative RNase protection assay. HGF, KGF, and EGF proteins were detected in rabbit lacrimal tissue using immunoprecipitation and western blot analysis. RESULTS: HGF mRNA and EGF mRNA were significantly increased in rabbit lacrimal gland tissue within 8 hours after corneal epithelial injury. The increase in KGF mRNA expression was small and reached significance I clay after corneal injury. Lacrimal gland expression peaked at 3 days after wounding for each growth factor mRNA, the same day, on average, that the epithelial defect healed. After the peak increase in expression, there was a progressive decline in expression of each growth factor mRNA, but production was still increased compared with prewound levels. HGF protein, KGF protein, and EGF proteins were detected in rabbit lacrimal gland tissue. CONCLUSIONS: Levels of HGF, KGF, and EGF mRNAs increase in rabbit lacrimal gland tissue in response to corneal epithelial wounding. The results of this study are consistent with the existence of a cornea-nervous system-lacrimal gland regulatory loop modulating expression of these growth factor mRNAs. The lacrimal gland is a likely source of increased HGF and EGF proteins detected in tears in previous studies. PMID- 10476783 TI - Experimental autoimmune keratitis induced in rats by anti-cornea T-cell lines. AB - PURPOSE: Idiopathic inflammation of the cornea, keratitis, has been proposed to result from an autoimmune process, but thus far no convenient animal model of keratitis exists. An attempt was made to establish an animal model for keratitis, to investigate possible autoimmune mechanisms. METHODS: T-cell lines were established from lymph node cells removed from rats immunized with bovine corneal epithelium (BCE) extract. After restimulation in vitro with BCE or a specific corneal antigen, the cells were transferred by intraperitoneal injection into naive rats, rats subjected to total body irradiation, or rats in which only one eye was irradiated. RESULTS: Neither direct immunization with corneal antigens nor transfer of activated anti-corneal T-cells into naive rats gave any signs of keratitis. Irradiation alone did not induce corneal inflammation. Transfer of corneal-specific activated T cells into irradiated rats produced keratitis starting around day 4 and culminating around day 8. The disease was self-limiting and the severity dependent on the dose and site of radiation. Keratitis was characterized by corneal haze, conjunctival and episcleral hyperemia, episcleral hemorrhages, chemosis, corneal infiltrates, and vascularization. Immunohistochemistry showed T-cell and macrophage infiltration of epithelium and stroma in the affected corneas. CONCLUSIONS: Thus, keratitis may be produced by T cells reactive to corneal antigens, provided that the target tissue has been made susceptible by irradiation. The effectiveness of T-cell vaccination in preventing adoptive keratitis suggests that systemic as well as local tissue factors may regulate the disease process. PMID- 10476784 TI - Interferon-gamma induces apoptosis and expression of inflammation-related proteins in Chang conjunctival cells. AB - PURPOSE: The purpose of this study was to investigate the effect of interferon (IFN)gamma on cell viability, cell growth, and apoptosis and on expression of apoptotic and inflammation-related proteins in epithelial conjunctival cells in vitro. Some aspects of transduction pathways of IFNgamma-induced alterations were also investigated, especially the role of protein kinase C (PKC) and IFNgamma transcriptional factor STAT1. METHODS: A human conjunctival cell line was treated with different concentrations (30 and 300 U/ml) of human recombinant IFNgamma. After 24, 48, and 72 hours of treatment, cell viability and relative cell number were studied with 3-(4,5-dimethylthiazol-2yl)2,5-diphenyl tetrazolium bromide (MTT) and crystal violet colorimetric assays. The apoptotic process was sought by phase-contrast microscopy, 4',6'-diamidino-2-phenylindole dihydrochloride (DAPI) staining, and transmission electron microscopy and was confirmed by DNA electrophoresis and immunoblotting of poly(ADP-ribose) polymerase (PARP). The cell cycle and expression of apoptotic proteins Fas, bax, and p53; of inflammation-related proteins HLA-DR and intercellular adhesion molecule (ICAM) 1; and of IFNgamma signal-transducing factor STAT1 were evaluated by flow cytometry and/or western blot analysis. To investigate PKC-related transduction pathways, two PKC modulators, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and staurosporine, were applied for 3 hours, followed by IFNgamma treatment for 72 hours. Moreover, the effects of PKC depletion were studied after a 24-hour application of TPA, also followed by IFNgamma treatment for 72 hours. Then, Fas, ICAM-1, and HLA-DR expressions were studied by flow cytometry. RESULTS: IFNgamma at 30 U/ml induced no change in cell cycle and in cell viability. Cell viability significantly decreased after 48 hours of treatment with 300 U/ml IFNgamma, associated with cell cycle alterations (decrease in number of cells in the S-M phase), apoptotic chromatin condensation and fragmentation, ladder pattern on DNA electrophoresis assay, and cleavage of PARP. Moreover, IFNgamma-treated cells overexpressed plasma membrane Fas, HLA-DR, and ICAM-1 in a dose- and time dependent manner, and STAT1 in both nuclear and cytosolic cell fractions. Only 300 U/ml IFNgamma-treated cells overexpressed bax, whereas Bcl-2 and p53 proteins were not modified. HLA-DR and Fas were upregulated after addition of staurosporine or after PKC-depleting treatment and repressed with TPA. Staurosporine, PKC depletion, and TPA all enhanced ICAM-1 expression. CONCLUSIONS: In our model, IFNgamma induced expression of inflammatory molecules and apoptotic mediators, cell growth arrest, and apoptosis of Chang conjunctival cells. Moreover, our results suggest that activation of PKC is not involved in some IFNgamma cellular effects that possibly imply the upregulation and nuclear translocation of STAT1. IFNgamma-induced apoptosis could explain in part the recently reported coexistence of inflammation and programmed cell death in ocular surface inflammatory disorders such as Sjogren's syndrome. PMID- 10476785 TI - On the role of kerato-epithelin in the pathogenesis of 5q31-linked corneal dystrophies. AB - PURPOSE: Recently, the authors identified a gene, BIGH3, in which different mutations cause a group of hereditary corneal dystrophies: lattice type I and IIIA (CDLI and CDLIIIA), granular Groenouw type I (CDGGI), Avellino (CDA), and Reis-Bucklers' (CDRB). All these disorders are characterized by the progressive accumulation of corneal deposits with different structural organization. Experiments were conducted to determine the role of kerato-epithelin (KE), the product of BIGH3, in the pathogenesis of the diseases. METHODS: KE-15 and KE-2, two rabbit antisera raised against peptides from the 69-364 and 426 - 682 amino acid regions of KE respectively, were used for immunohistology of the corneas obtained after keratoplasty in six CDLI patients, three CDGGI patients, and one CDA patient. RESULTS: The nonamyloid deposits observed in CDGGI stained intensively with KE-15 and KE-2, whereas the amyloid deposits in all analyzed CDLI corneas reacted to KE-2 but not to KE-15. In the CDA cornea, where amyloid and nonamyloid inclusions were present, positive staining with both antisera was observed. CONCLUSIONS: Pathologic amyloid and nonamyloid deposits observed in CDLI, CDGGI-, and CDA-affected corneas are caused by KE accumulation. Different staining patterns of amyloid and nonamyloid deposits observed with antibodies against the amino and carboxyl termini of KE suggest that two mechanisms of KE misfolding are implicated in the pathogenesis of 5q31-linked corneal dystrophies. PMID- 10476786 TI - Secretory peptides TFF1 and TFF3 synthesized in human conjunctival goblet cells. AB - PURPOSE: The objective of this study was to determine whether human conjunctival epithelium synthesizes TFF-peptides (formerly P-domain peptides, trefoil factors), a family of mucin-associated secretory peptides of the gastrointestinal tract. METHODS: Expression of TFF-peptides in human conjunctiva was monitored by reverse transcription-polymerase chain reaction analysis. Antisera specific for TFF-peptides were used for immunohistochemistry to determine the presence and distribution of TFF-peptides in human conjunctiva. RESULTS: mRNA expression of TFF1 and TFF3, but not TFF2, was detected in human conjunctiva. TFF1 and TFF3 but not TFF2 are stored in conjunctival goblet cells only as revealed by immunofluorescence. CONCLUSIONS: Goblet cells of the human conjunctiva synthesize TFF1 and TFF3. These peptides, together with the secretory ocular mucin MUC5AC, may contribute to the rheological properties of the tear film. They also may influence healing of corneal wounds due to their motogenic properties. PMID- 10476787 TI - Human anti-transforming growth factor-beta2 antibody: a new glaucoma anti scarring agent. AB - PURPOSE: Currently available anti-scarring regimens for glaucoma filtration surgery have potentially blinding complications and thus the need for alternative and safer agents. The effects of a new antibody to transforming growth factor (TGF)-beta2 on in vitro and in vivo conjunctival scarring and after glaucoma filtration surgery were investigated. METHODS: The activity of a novel recombinant monoclonal neutralizing antibody (mAb) to human TGF-2 (rhAnti-TGF beta2 mAb) was studied in conjunctival fibroblast-mediated proliferation, migration, and collagen contraction. Its safety in subconjunctival administration was assessed in vivo, and, in a rabbit model of glaucoma filtration surgery, its effects on conjunctival scarring and filtration surgery outcome were investigated. RESULTS: The rhAnti-TGF-beta2 mAb effectively inhibited TGF-beta2 mediated conjunctival scarring activity in vitro, at 50% inhibitory concentrations (IC50) of less than 1 nM. It significantly improved glaucoma filtration surgery outcome in an animal model of aggressive conjunctival scarring compared with control (P = 0.0291) and was clinically safe, nontoxic, and well tolerated after subconjunctival administration. CONCLUSIONS: Subconjunctival rhAnti-TGF-beta2 mAb treatment significantly affects surgical outcome and effectively reduces conjunctival scarring both in vitro and in vivo. It appears safe for subconjunctival administration and when compared with mitomycin-C treatment histologically, much less destructive to local tissue. rhAnti-TGF-beta2 mAb may have potential as a new anti-scarring agent for use in glaucoma filtration surgery. PMID- 10476788 TI - AlphaB-crystallin in the trabecular meshwork is inducible by transforming growth factor-beta. AB - PURPOSE: Because in glaucomatous eyes transforming growth factor-beta (TGF-beta) and alphaB-crystallin are increased in the anterior eye segment, the effect of TGF-beta1 and TGF-beta2 on the expression of alphaB-crystallin and its corresponding mRNA was studied in human trabecular meshwork (TM) cells. METHODS: Monolayer cultures of "cribriform" and "corneoscleral" TM cells of 5 human donors (12-73 years of age) were treated with either 1.0 ng/ml TGF-beta1, TGF-alpha2, or 5 X 10(-7) dexamethasone (DEX) for 12 to 96 hours. Induction of alphaB-crystallin and the related mRNA was investigated by western and northern blot analyses. For comparison, human foreskin fibroblasts (HFF) and NIH 3T3 cells were treated in the same way as the TM cells. RESULTS: An increase of alphaB-crystallin mRNA was observed after treatment of TM cells with TGF-beta1 and TGF-beta2, whereas DEX had no effect. In the cribriform TM cells with a high basal level, the enhancement ranged between 2 and 3 times; whereas in the corneoscleral TM cells alphaB-crystallin mRNA increased between 5 and 6 times. Using western blot analysis, the increase of alphaB-crystallin expression in the cribriform TM cells was only small compared with the significant increase in the corneoscleral TM cells. Treatment of HFF and NIH 3T3 cells with TGF-beta did not induce alphaB crystallin mRNA. CONCLUSIONS: This is the first time to show that alphaB crystallin is not only induced by stress factors but also by TGF-beta in TM cell cultures. The difference in induction of mRNA and protein seems to be dependent on alphaB-crystallin concentration before treatment. PMID- 10476789 TI - Ganglion cell losses underlying visual field defects from experimental glaucoma. AB - PURPOSE: To investigate the relationship between ganglion cell losses and visual field defects caused by glaucoma. METHODS: Behavioral perimetry and histology data were obtained from 10 rhesus monkeys with unilateral experimental glaucoma that was induced by argon laser treatments to their trabecular meshwork. After significant visual field defects had developed, the retinas were collected for histologic analysis. The ganglion cells were counted by light microscopy in cresyl violet-stained retina sections, and the percentage of ganglion cell loss (treated to control eye counts) was compared with the depth of visual field defect (treated to control eye thresholds) at corresponding retinal and perimetry test locations. Sensitivity losses as a function of ganglion cell losses were analyzed for Goldmann III, white and Goldmann V, and short- and long-wavelength perimetry test stimuli. RESULTS: The relationship between the proportional losses of ganglion cells and visual sensitivity, measured with either white or colored stimuli, was nonlinear. With white stimuli, the visual sensitivity losses were relatively constant (approximately 6 dB) for ganglion cell losses of less than 30% to 50%, and then with greater amounts of cell loss the visual defects were more systematically related to ganglion cell loss (approximately 0.42 dB/percent cell loss). The forms of the neural-sensitivity relationships for visual defects measured with short- or long-wavelength perimetry stimuli were similar when the visual thresholds were normalized to compensate for differences in expected normal thresholds for white and colored perimetry stimuli. CONCLUSIONS: Current perimetry regimens with either white or monochromatic stimuli do not provide a useful estimate of ganglion cell loss until a substantial proportion have died. The variance in ganglion cell loss is large for mild defects that would be diagnostic of early glaucoma and for visual field locations near the fovea where sensitivity losses occur relatively late in the disease process. The neural sensitivity relationships were essentially identical for both white and monochromatic test stimuli, and it therefore seems unlikely that the higher sensitivity for detecting glaucoma with monochromatic stimuli is based on the size-dependent susceptibility of ganglion cells to injury from glaucoma. PMID- 10476790 TI - Risk factors for late presentation in chronic glaucoma. AB - PURPOSE: To identify the risk factors for having advanced glaucomatous visual field loss on the first visit at three hospital eye services. METHODS: This was a hospital-based, case-control study involving patients newly diagnosed with glaucoma at first visit to one of three ophthalmic departments in the United Kingdom. Patients with a previous history of ocular hypertension or any documented suspicion of glaucoma (within the hospital eye service) were excluded. RESULTS: Occupational group, initial intraocular pressure (IOP), family history of glaucoma, method of referral to hospital, and the number of years since the last visit to an optometrist were found to be independently associated with late presentation. A linear trend of increasing odds of late attendance was associated with increasing Standard Occupational Classification. Those in managerial (category II) and skilled (category III) groups estimated (95% confidence intervals) to be, respectively. 0.2 (0.00, 0.16) and 0.27 (0.1, 0.8) as likely to attend with advanced glaucomatous field loss as unskilled (category V) people with similar initial IOP, family history, referral route, and time since last optometrist visit. The data strongly suggest an association between IOP and advanced field loss at initial hospital examination. There was a 1.2 (1.12, 1.28) increase in the OR of late presentation per unit increase in millimeters of mercury after adjustment for the other mentioned factors. People with a family history of glaucoma were estimated to be almost one third (adjusted OR, 0.29 [0.12, 0.74]) as likely to have advanced field loss as those with no family history. People referred by any source other than an optometrist who has made the correct diagnosis of glaucoma were 4.5 times (adjusted OR, 4.53 [1.52, 13.48]) more likely to be late attenders than patients so referred but similar in other mentioned factors. These data also provide strong evidence that the more years since the last visit to an optometrist, the greater the likelihood of having advanced glaucomatous visual field loss on the first visit to the eye service (adjusted OR per year, 1.25 [1.10, 1.42]). CONCLUSIONS: These data strongly suggest that certain subgroups of people with glaucoma were at greater risk of having advanced and irremediable field loss on first visiting the eye services studied. PMID- 10476791 TI - Fas-Fas ligand-mediated apoptosis within aqueous during idiopathic acute anterior uveitis. AB - PURPOSE: Despite ocular immune privilege, (auto)immune-mediated acute anterior uveitis (AAU) is relatively common. However, although relapses of AAU are usually self-limiting, possible regulatory mechanisms remain undefined in humans. Experimentally, Fas-Ligand (FasL)-mediated apoptosis of Fas+ inflammatory cells contributes to the immune privilege within the anterior chamber and provides an explanation for the success of corneal allograft transplantation. Therefore, whether such mechanisms regulate the immune response in AAU was investigated. METHODS: Aqueous and peripheral blood samples from consecutive patients presenting with idiopathic AAU were obtained with consent. Leukocytic phenotype was analyzed by flow cytometry, and apoptosis was determined by both flow cytometry and TdT-dUTP terminal nick-end labeling analysis. Presence of soluble Fas and FasL was determined by western blot analysis and enzyme-linked immunosorbent assay and compared with control aqueous from patients undergoing cataract surgery. The ability of the aqueous to induce apoptosis in a Fas+ Jurkat cell line was also determined. RESULTS: During AAU aqueous-infiltrating Fas+ cells included CD3+ T cells and granulocytes, whereas FasL+ cells comprised predominantly of non-CD3+ T cells. Higher levels of functional soluble FasL were found in aqueous of AAU patients than in normal aqueous, capable of inducing apoptosis in 68.9% +/- 7.6% of Fas+ lymphoid cells. Compared with peripheral blood, the CD4+ T cells infiltrate within aqueous showed significantly increased CD69 and CD25(IL-2r) expression. Flow cytometric analysis of aqueous showed that 9.32% +/- 1.2% of infiltrating non-granulocyte CD45+ cells were apoptotic, confirmed as T cells on subsequent three-color flow cytometric analysis. CONCLUSIONS: Taken together with published experimental data, the present study provides evidence for FasL-mediated apoptotic cell death contributing to the local immune regulation of ocular inflammatory disease and provides a mechanism to account for the self-limiting clinical course of AAU. PMID- 10476792 TI - Specific aqueous humor factors induce activation of regulatory T cells. AB - PURPOSE: To examine the possibility that aqueous humor-induced regulatory T cells could function in vivo, these T cells were examined for their ability to suppress adoptive transfer of delayed-type hypersensitivity (DTH). To begin to understand the mechanisms by which aqueous humor induces activation of regulatory T cells, alpha-melanocyte stimulating hormone (MSH) and transforming growth factor (TGF) beta2 were examined for ability to induce regulatory T cells. METHODS: Primed T cells were treated with aqueous humor and assayed for regulatory activity by injecting them intravenously along with DTH-mediating T cells into syngeneic mice. Antigen-pulsed antigen-presenting cells (APCs) were injected into the pinna of the mouse ear, and swelling was measured 24 hours later. Primed T cells were also activated in vitro in the presence of alpha-MSH, TGF-beta1, or TGF-beta2 and were assayed for proliferation and TGF-beta production along with suppressing DTH. RESULTS: Aqueous humor-treated T cells suppressed inflammation mediated by DTH T cells. Maximum regulatory T cell activity was induced when primed T cells were activated in vitro in the presence of alpha-MSH followed 4 hours later with active TGF-beta2. Such T cells proliferated, produced TGF-beta, and suppressed DTH, suggesting that alpha-MSH and TGF-beta2 induce activation of regulatory T cells. No regulatory T cell activity could be induced in the presence of TGF beta1. CONCLUSIONS: The ocular microenvironment constitutively produces immunoregulatory factors that suppress the induction of inflammatory activity and promotes regulatory T cell activity. Such regulatory T cells can further contribute to maintaining the normal immunosuppressive ocular microenvironment through their ability to suppress activation of other inflammatory T cells. PMID- 10476793 TI - Inhibition of endotoxin-induced uveitis and potentiation of local TNF-alpha and interleukin-6 mRNA expression by interleukin-13. AB - PURPOSE: To investigate the effect of systemic injections of interleukin (IL)-13 on the development of endotoxin-induced uveitis (EIU) in the rat. METHODS: EIU was induced in Lewis rats by a single footpad injection of lipopolysaccharide (LPS; 350 microg/kg) from Salmonella typhimurium. Rats were treated with a subcutaneous injection in the back of recombinant human IL-13 (50 microg/kg in 0.2 ml of saline) performed 30 minutes before LPS injection and 6 and 10 hours afterward. At 23 hours after LPS injection, EIU was evaluated by slit-lamp examination and by counts of inflammatory cells on cryostat sections after specific immunostaining. The expression of nitric oxide synthase (NOS)-II in ocular tissues was determined by dual immunofluorescent staining and the release of nitrite in aqueous humor by Griess reaction. Cytokine gene expression in the iris/ciliary body, choroid, and retina was evaluated by reverse transcription polymerase chain reaction. RESULTS: At 24 hours after LPS injection, significant clinical inhibition of ocular inflammation and fibrin deposition in the eye was observed in IL-13-treated rats. Quantitative analysis of ocular tissues revealed a significant decrease of OX42+ cells (microglia, activated macrophages, dendritic cells, and polymorphonuclear leukocytes) and ED-1+ cells (monocytes/macrophages and dendritic cells). No effect on ED2+ cells (resident tissue macrophages) was found. Treatment with IL-13 decreased nitrite levels in aqueous humor and enhanced the expression of tumor necrosis factor-alpha (TNF alpha) and IL-6 mRNA in ocular tissues. CONCLUSIONS: Interleukin-13 treatment inhibits LPS-induced ocular inflammation with inhibition of nitrite release and increased TNF and IL-6 production in the eye. These results confirm the role of the NO pathway in the pathogenesis of EIU and suggest the involvement of TNF and IL-6 in the downregulation of ocular inflammation. PMID- 10476794 TI - The role of antibody to human beta4 integrin in conjunctival basement membrane separation: possible in vitro model for ocular cicatricial pemphigoid. AB - PURPOSE: To demonstrate the specific binding of autoantibodies present in the sera of patients with ocular cicatricial pemphigoid (OCP) to human beta4 integrin present in the normal human conjunctiva (NHC) and to study the role of OCP autoantibodies and antibody to human beta4 integrin in the pathogenesis of subepithelial lesion formation in OCP. METHODS: Indirect immunofluorescence assay and in vitro organ culture method using NHC were used. Sera and IgG fractions from 10 patients with OCP; immunoaffinity-purified OCP autoantibody; antibodies to human beta4, beta1, alpha6, and alpha5 integrins; and sera from patients with pemphigus vulgaris, bullous pemphigoid (BP), and chronic atopic and chronic ocular rosacea cicatrizing conjunctivitis; and normal human serum (NHS) were used. RESULTS: Nine of 10 OCP sera or IgG fractions, immunoaffinity-purified OCP autoantibody, antibodies to human beta4 and alpha6 integrins, and sera from patients with BP showed homogenous, smooth linear binding along the basement membrane zone (BMZ) of the NHC. NHS, antibodies to other integrins, and sera from patients with chronic cicatrizing conjunctivitis from other causes showed no such binding. When NHC was first absorbed with OCP sera and then reacted with anti beta4 antibodies or vice versa, the intensity of the BMZ binding was dramatically reduced or completely eliminated, indicating that there were autoantibodies in OCP sera specific for the beta4 integrin. BMZ separation developed 48 to 72 hours after addition of total OCP sera, IgG fractions from OCP sera, immunoaffinity purified autoantibodies from sera of patients with OCP, or anti-beta4 antibodies to the NHC cultures, but not after addition of normal control sera, sera from patients with chronic cicatrizing conjunctivitis from causes other than OCP, or sera from patients with OCP in clinical remission. CONCLUSION: Circulating anti beta4 integrin antibody may have an important role in the pathogenesis of OCP. PMID- 10476795 TI - Lens epithelium and fiber Na,K-ATPases: distribution and localization by immunocytochemistry. AB - PURPOSE: To use immunofluorescence and immunogold techniques to identify the catalytic subunits of the Na,K-ATPases of the lens and to determine their location in the cells of the epithelium and cortex of bovine and human lenses. METHODS: Frozen sections of capsulated and decapsulated bovine and human lenses were prepared, blocked, and treated with affinity-purified polyclonal rabbit antibodies to the Na,K-ATPase catalytic subunit isoforms with subsequent treatment with fluorescein isothiocyanate-labeled goat anti-rabbit IgG and visualization of the fluorescence by light microscopy. An immunogold-labeled goat anti-rabbit IgG was used to detect, by electron microscopy, the binding of the same affinity-purified polyclonal antibodies to thin sections of decapsulated lenses that had been fixed and embedded in Lowicryl K4M. The results were confirmed by staining of western blot analysis of sodium dodecyl sulfate polyacrylamide gel separations of enriched membrane preparations from bovine and human lenses. RESULTS: The three common catalytic subunits of the Na,K-ATPases are present in the plasma membranes of lens epithelium, lens fibers, or both. The data indicate a polarized distribution of the alpha1 and alpha3 catalytic subunit isoforms in central epithelium. In the cortical fibers, the alpha2 isoform is present around the interdigitations. The alpha3 isoform is found in the interdigitation-free regions of human cortical fibers. CONCLUSIONS: This unique distribution of Na,K-ATPases precludes the popular pump-leak model for lens monovalent cation homeostasis. The functional significance of the distribution of Na,K-ATPases in the lens epithelium and superficial fibers is currently under investigation. PMID- 10476796 TI - Characterization of alpha2 adrenergic receptor subtypes in human ocular tissue homogenates. AB - PURPOSE: To determine the predominant alpha2 adrenergic receptor subtypes present in the human eye. METHODS: Saturation- and competition-receptor- binding experiments were performed with the radioligand [3H]RX821002 in human ciliary body, retinal pigmented epithelium-choriocapillaris, iris, and neurosensory retina. The affinities of various adrenergic antagonists in these ocular tissues were compared with their affinities for the cloned alpha2A, alpha2B, and alpha2C adrenergic receptor subtypes. RESULTS: The density of alpha2 adrenergic receptors was highest in the iris (440 femtomoles/mg protein), lowest in the neurosensory retina (14 femtomoles/mg protein), and intermediate in the other two tissues (approximately 90 fmol/mg protein). The drug affinities in all four human ocular tissues were highly correlated (correlation coefficients between 0.94 and 0.97) with the affinities for the human alpha2A adrenergic receptor subtype and poorly correlated (correlation coefficients between 0.15 and 0.66) with the alpha2B and alpha2C subtypes. CONCLUSIONS: In agreement with previous studies in several animal species, the alpha2 adrenergic receptors in the human ciliary body, retinal pigmented epithelium-choriocapillaris, iris, and neurosensory retina are predominately of the alpha2A subtype. PMID- 10476797 TI - Light and choroidal PO2 modulation of intraretinal oxygen levels in an avascular retina. AB - PURPOSE: To determine the influence that choroidal oxygen level and outer retinal oxygen demand have on oxygen availability to the inner retina when the choroid is the only source of retinal oxygenation. This condition prevails in avascular retinas and in vascularized retinas suffering vascular occlusion. METHODS: Oxygen sensitive microelectrodes were used to measure the oxygen tension as a function of depth in the naturally avascular retina of anesthetized and mechanically ventilated guinea pigs (n = 6). Choroidal PO2 was manipulated by varying the ventilation gas mixture, and outer retinal oxygen consumption was modulated by light-dark adaptation. Individual PO2 profiles were fitted to a multilayer mathematical model of PO2 distribution, and pairs of profiles at different choroidal PO2 levels, or under light and dark conditions, were fitted to an intraretinal PO2 difference model. Both models reflect the purely choroidal supply of retinal oxygenation. RESULTS: An increase in choroidal PO2 produced an equivalent increase in all retinal layers. Light induced a decreased oxygen consumption in the region of the inner segments of the photoreceptors, which resulted in a significant increase in PO2 in this layer, flowing on unattenuated to all inner retinal layers. The intraretinal PO2 distribution and the light- and ventilatory-induced changes in PO2 were consistent with theoretical predictions of the mathematical models. CONCLUSIONS: The present experimental studies confirm that when the choroid is the only source of retinal oxygenation, the full effect of increased choroidal oxygen level or reduced uptake in the outer retina passes through to the inner retinal layers if the oxygen utilization by the inner retina remains constant. PMID- 10476798 TI - Ultrastructural localization of light-induced lipid peroxides in the rat retina. AB - PURPOSE: Localization of light-induced lipid peroxides in the rat retina at an ultrastructural level as benzidine-reactive substances. METHODS: Long-Evans rats with nondilated pupils were exposed to intense light of 6000 lux for 12 or 24 hours. Control animals were kept under physiological light conditions. Rats with dilated pupils were exposed to a light intensity of 50 lux or 150,000 lux for 1 hour. For ultrastructural localization the enucleated eyes were fixed in a 0.1-M cacodylate buffer (pH 7.4) containing 2% glutaraldehyde for 2 hours. Pieces of the superior part of the central eyecup were incubated overnight with tetramethylbenzidine (TMB; pH 3.0) at 4 degrees C, postfixed with 1.5% OSO4, and embedded for electron microscopy. RESULTS: In animals exposed to intense light, electron-dense structures appeared exclusively throughout the rod outer segments after an irradiation of 6000 lux for 24 hours or 150,000 lux for 1 hour and were absent in animals with nondilated pupils kept at physiological light conditions. Dilation of the pupils leads to the appearance of electron-dense structures after just 1 hour of 50 lux, whereas rats with nondilated pupils withstand even a 12 hour irradiation with 6000 lux. No electron-dense structures were found when no TMB was used in incubation. CONCLUSIONS: The appearance of electron-dense structures in the rod outer segments depends on the incubation with TMB and intensive light exposure of the rat. Dilation of the pupils lowers the threshold for the emergence of electron-dense structures significantly. This strongly supports the view that light-induced lipid peroxides in the rat retina are localized at an ultrastructural level as benzidine-reactive substances. This protocol presents a tool for the generation and ultrastructural localization of lipid peroxides in rat retinas. PMID- 10476799 TI - Effects of photodynamic therapy using verteporfin on experimental choroidal neovascularization and normal retina and choroid up to 7 weeks after treatment. AB - PURPOSE: To study the long-term effects of photodynamic therapy (PDT), using liposomal benzoporphyrin derivative (BPD) or Verteporfin, on experimental choroidal neovascularization (CNV) and on normal retina and choroid (with no CNV) in the cynomolgus monkey eye. METHODS: Photodynamic therapy was performed in 8 cynomolgus monkey eyes with experimental CNV induced by laser injury. The effect of PDT on normal retina and choroid (with no CNV) was studied in 9 monkey eyes. Liposomal BPD was administered intravenously (0.375 mg/kg) either as a bolus, as a slow infusion over 32 minutes, or as a fast infusion over 10 minutes. Photodynamic therapy was performed using light at a wavelength of 689 or 692 nm, with an irradiance of 600 mW/cm2 and fluence of 150 J/cm2. Follow-up studies, including fundus photography and FA, were performed at 24 hours after PDT and then weekly. Indocyanine green and BPD angiography were performed in selected cases. Tissues were examined with light and electron microscopy at the end of follow-up. RESULTS: Twenty-three of the 32 areas of CNV treated with PDT showed absence of angiographic leakage at 24 hours. Twenty-eight areas of CNV were followed for 4 weeks; 22 of 28 showed absence of angiographic leakage at 2 weeks; and 20 of 28 at 4 weeks of follow-up. Forty spots on the normal retina and choroid were treated with PDT and were followed for 4 to 7 weeks. These spots showed pigment-laden cells in the outer retina, variably pigmented retinal pigment epithelium (RPE) in the treated area, intact neurosensory retina, and reperfusion of the choriocapillaris. CONCLUSIONS: Photodynamic therapy leads to absence of angiographic leakage for at least 4 weeks in experimental CNV in the monkey model. In the normal monkey eye the RPE and choriocapillaris show generalized recovery with preservation of the neurosensory retina 7 weeks after PDT. PMID- 10476800 TI - The interpretation of optical coherence tomography images of the retina. AB - PURPOSE: To determine the relationship between optical coherence tomography (OCT) images of the retina and retinal substructure in vitro and in vivo. METHODS: In vitro, OCT images of human and bovine retina were acquired after sequential excimer laser ablation of the inner retinal layers. Measurements of bands in the OCT images were compared with measurements of retinal layers on histology of the ablated specimens. In vivo, OCT images were acquired of retinal lesions in which there was a displacement of pigmented retinal pigment epithelial (RPE) cells: retinitis pigmentosa and laser photocoagulation (eight eyes each). RESULTS: The mean thickness of human inner OCT bands (131 microm; 95% confidence interval [CI], 122-140 microm) was 7.3 times that of the retinal nerve fiber layer (RNFL). This band persisted despite ablation greater than 140 microm. The inner aspect of the outer OCT band corresponded to the apical RPE, but the mean thickness of this band in human tissue (55 microm; 95% CI, 48-62 microm) was 2.6 times the thickness of the RPE-choriocapillaris complex. OCT measurement of total retinal thickness was accurate (coefficient of variance, 0.05) and precise (coefficient of correlation with light microscopy, 0.98). Hyperpigmented lesions gave rise to high signal, attenuating deeper signal; hypopigmented lesions had the opposite effect on deeper signal. CONCLUSIONS: The inner band is not RNFL specific, partly consisting of a surface-related signal. The location, not thickness, of the outer band corresponds to RPE melanin. Given the additional effect of polarization settings, precise OCT measurement of specific retinal layers is currently precluded. PMID- 10476801 TI - Thresholds for visible lesions in the primate eye produced by ultrashort near infrared laser pulses. AB - PURPOSE: To evaluate the effects of near-infrared (near-IR) ultrashort laser pulses on the retinas of rhesus monkey eyes and to perform threshold measurements for minimum visible lesions (MVLs) at pulse widths ranging from nanoseconds to femtoseconds. METHODS: Near-infrared single laser pulses were placed within the macular area of live rhesus monkey eyes for five different pulse widths (7 nsec; 80, 20, and 1 psec; and 150 fsec). One visible wavelength of 530 nm at 100 fsec was also included in the study. Visible lesion thresholds (MVL-ED50) were determined 1 hour and 24 hours after exposure. Fluorescein angiography thresholds (FAVL-ED50) were also determined using a probit analysis of the dosage. Thresholds were calculated as that dosage causing a 50% probability for damage, and the fiducial limits were calculated at the 95% confidence level. RESULTS: For all pulse widths, the 24-hour MVL-ED50 was lower than the 1-hour MVL-ED50, and they both decreased with decreasing pulse width. Thresholds at the 1-hour reading decreased from 28.7 microJ at 7 nsec to 1.8 microJ at 150 fsec, whereas thresholds at 24 hours decreased from 19.1 microJ at 7 nsec to 1.0 microJ at 150 fsec. The doubled 1060-nm wavelength of the 530-nm threshold decreased from 0.36 to 0.16 microJ after 24 hours. FAVL-ED50s were much higher than MVL-ED50s, showing that FA was not as sensitive in determining damage levels. CONCLUSIONS: Laser pulse widths less than 1 nsec in the near-IR are capable of producing visible lesions in rhesus monkey eyes with pulse energies between 5 and 1 microJ. Also, the near-IR thresholds for these pulse widths are much higher than for the visible wavelengths. As with visible wavelengths, FA is not as sensitive in determining threshold levels as is visually observing the retina through a fundus camera. PMID- 10476802 TI - Identification of a nervous tissue-specific chondroitin sulfate proteoglycan, neurocan, in developing rat retina. AB - PURPOSE: To identify the expression of neurocan, a nervous tissue-specific chondroitin sulfate proteoglycan, in retina and to elucidate its changes during development. METHODS: Expressional changes of neurocan mRNAs in developing rat retinas were investigated by a semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). The localization and characterization of neurocan core proteins were also investigated with the use of Western blot analysis and immunohistochemistry. RESULTS: Gene expression of neurocan was identified in retinas by RT-PCR. Semiquantitative analysis using Southern blot analysis revealed that mRNA expression for neurocan increased at increasing postnatal stages and that it reached its peak around postnatal day 7 (P7). Immunohistochemical studies demonstrated that in differentiating rat retinal (neuroblast) cells weak neurocan immunoreactivities were observed throughout the retina on embryonal days 14 (E14) and E16. During the early postnatal period, the immunoreactivities became most conspicuous in the inner and outer plexiform layers on P7 through P14. In adult retinas, only faint immunostaining was detected. Immunoblot analysis showed two positive bands of 220- and 150-kDa core glycoproteins after treatment with chondroitinase ABC. Further immunoblot analysis revealed that the expression of these two immunolabeled variants was regulated differently during retinal development. CONCLUSIONS: The temporal and spatial regulation of expression of neurocan and its proteolytic variant during retinal development suggest that it may play a role in differentiation and neural network formation. PMID- 10476803 TI - Survival of axotomized retinal ganglion cells in peripheral nerve-grafted ferrets. AB - PURPOSE: Peripheral nerve (PN) grafting to the optic nerve stump stimulates not only axonal regeneration of the axotomized retinal ganglion cells (RGCs) into the grafted PN but also their survival. The purpose of the present study was to determine the number, distribution, and soma diameter of only surviving RGCs without regenerated axons and surviving RGCs with regenerated axons in PN-grafted mammals. METHODS: A segment of PN was grafted to the optic nerve stump of adult ferrets. Two months after the PN grafting, surviving RGCs with regenerated axons were retrogradely labeled with granular blue (GB) and stained with RGC-specific antibody C38. Surviving RGCs without regenerated axons were identified as C38 positive cells without GB labeling. RESULTS: Twenty-one percent of RGCs survived axotomy after PN grafting in the area centralis (AC), whereas 47% survived in the peripheral retina. Twenty-six percent of surviving RGCs in the AC exhibited axonal regeneration, which was higher than that in the peripheral retina. Soma diameter histograms revealed that RGCs with regenerated axons showing both GB and C38 positivity were in the large soma diameter ranges. In contrast, the soma diameter distribution of surviving RGCs that did not have regenerated axons showed a peak in the smaller soma diameter ranges. CONCLUSIONS: The present data suggest that PN grafting promotes survival of axotomized RGCs more effectively in the peripheral retina than in the AC. Among surviving RGCs, the larger cells exhibited axonal regeneration into the grafted PN, whereas the axons of smaller cells did not to regenerate in either the AC or the peripheral retina. PMID- 10476804 TI - TIMP-3 in Bruch's membrane: changes during aging and in age-related macular degeneration. AB - PURPOSE: To assess the distribution, content, and function of tissue inhibitor of metalloproteinases (TIMP)-3 during aging in normal eyes for comparison with the levels observed in eyes with age-related macular degeneration (AMD). METHODS: Donor tissues analyzed included 36 normal eyes (14-96 years old) and 15 AMD eyes (74 -98 years old). A tissue strip including the fovea was used for immunohistochemistry. Western blot analysis was performed on extracts of the retinal pigment epithelium (RPE)- choroid complex from the posterior part of each eye. Immunoreactivity of TIMP-3 bands in each western blot was densitometrically quantitated. The inhibitory function of TIMP-3 was evaluated with reverse zymography. RESULTS: TIMP-3 was present uniformly across Bruch's membrane in the normal samples. In samples from donors more than 50 years of age, immunostaining was intense. TIMP-3 content ranged from 92 to 1061 ng/cm2 and increased with age (r = 0.66). In AMD eyes, TIMP-3 distribution in Bruch's membrane was abundant in areas of continuous soft drusen but absent in areas below RPE atrophy. TIMP-3 levels in AMD eyes were significantly higher than in age-matched normal eyes (577 versus 877 ng/cm2; P = 0.009). Inhibitory activity correlated well with TIMP-3 content (r = 0.82) and was also significantly higher in AMD eyes than in age matched normal eyes (P < 0.001). CONCLUSIONS: During normal aging, TIMP-3 content in Bruch's membrane of the macula shows a significant increase. TIMP-3 content in AMD eyes was elevated relative to that of age-matched normal eyes. Higher levels of TIMP-3 may contribute to the thickening of Bruch's membrane observed in AMD. PMID- 10476805 TI - Immunohistochemical localization of somatostatin receptor subtypes sst1 and sst2 in the rat retina. AB - PURPOSE: To investigate the distribution of somatostatin receptor subtypes sst1 and sst2 in the rat retina by immunohistochemistry and to characterize further the neurotransmitters of the sst1- and sst2-immunoreactive cells. METHODS: Polyclonal antibodies raised against sst1 and sst2 receptors were applied to 12 microm cryostat sections of rat retinas fixed in paraformaldehyde. Further, immunofluorescence double labeling was performed for the sst1 and sst2 receptors with somatostatin, tyrosine hydroxylase (TH) and glutamate decarboxylase (GAD). RESULTS: Immunoreactivity for sst1 was present in somatostatinergic amacrine cells located in the inner nuclear layer (INL) and in displaced amacrine cells in the ganglion cell layer of the retina. Also, a small number of ganglion cells were sst1 immunoreactive. Immunoreactivity for sst2 was observed in many medium sized amacrine cells in the middle part of the INL, with a central process projecting to the sublaminae of the inner plexiform layer. Furthermore, sst2 immunoreactivity was found in large amacrine cells of the INL. These cells also contained TH. Inner segments of cone receptors were stained with the sst2 antiserum. Immunostaining for sst2, and to a minor extent for sst1, was found in Muller cell fibers. None of the somatostatin receptors colocalized with GAD. CONCLUSIONS: These findings suggest that the sst1 receptor may function as an autoreceptor on retinal somatostatinergic cells. The presence of sst2 receptors on the TH-immunoreactive amacrine cells indicates an influence of somatostatin on the secretion of dopamine in rat retina. PMID- 10476806 TI - Light treatment enhances photoreceptor survival in dystrophic retinas of Royal College of Surgeons rats. AB - PURPOSE: To determine whether treatment with bright light elicits a protective response that enhances photoreceptor survival in Royal College of Surgeons (RCS) rats with inherited retinal degeneration. METHODS: RCS rats were illuminated for 10 to 12 hours with 130 foot-candles (fc) of white or green light. Untreated littermates that were kept under low cyclic light levels were used as control subjects. Photoreceptor survival was determined by quantitative analysis of photoreceptor nuclei and ultrastructural assessment of cellular organization. Basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF) gene expression were determined at the mRNA and protein levels. RESULTS: Treatments of RCS rats with a single dose of bright light on postnatal day 23 (P23) greatly enhanced photoreceptor survival. Ultrasturctural analysis revealed intact inner segments in light-treated retinas, whereas in untreated retinas only remnants of inner segments were observed. By P42, numerous viable nuclei were counted in the posterior retina of light-treated rats, whereas most of the remaining nuclei in untreated RCS rat retinas were highly pyknotic. At 2.5 days after treatment with a single dose of bright light, bFGF gene expression was significantly higher than in untreated RCS rat retinas. By P42, bFGF protein levels were still significantly higher in the treated retinas. CONCLUSIONS: Exogenous bFGF has been shown to promote photoreceptor survival in the RCS rat retina. Thus, the increased bFGF expression that was measured in the light-treated RCS rat retinas may be a protective response to light stress, which supports the observed rescue of photoreceptors in light-treated RCS rat retinas. PMID- 10476807 TI - N-methyl-D-aspartate (NMDA)--induced apoptosis in rat retina. AB - PURPOSE: The involvement of apoptosis in N-methyl-D-aspartate (NMDA)-induced excitotoxicity in adult rat retinas was examined. METHODS: Excitotoxic loss of inner retinal elements was induced by intravitreal injections of various concentrations of neutralized NMDA in adult albino Lewis rats. Tissue responses were quantified by measuring the inner retinal thickness (IRT) in plastic sections of the retinas and cell counts in the retinal ganglion cell layer in flatmount preparations of the whole retinas. Internucleosomal DNA fragmentation, a hallmark of apoptosis, was assayed with agarose DNA gel electrophoresis. The in situ TdT-mediated biotin-dUTP nick end labeling (TUNEL) method was used to locate nicked DNA in paraffin sections of the retinas. Ultrastructural changes of the degenerating cells were examined by electron microscopy. The efficacy of Ac-Tyr Val-Ala-Asp-CMK (YVAD-CMK), a peptidyl caspase inhibitor, and 3-aminobenzamide (ABA), an inhibitor of poly(ADP-ribose) polymerase (PARP), in ameliorating the loss of inner retinal elements was evaluated using morphometry to examine the apoptotic pathways. RESULTS: Intravitreal injection of NMDA induced a dose dependent loss of inner retinal elements as evidenced by the measurements of IRT and RGCCs. There were time- and dose-related appearances of internucleosomal fragmentation of retinal DNA and a time-related appearance of TUNEL-positive nuclei in the inner retinas after intravitreal NMDA injection. Ultrastructural features consistent with classic apoptotic changes were noted in degenerating cells in the retinal ganglion cell layer and the inner nuclear layer. Control retinas given vehicle, N-methyl-L-aspartate (the L-isomer of NMDA), or NMDA plus MK-801, a specific antagonist, did not show these changes. Simultaneous administration of NMDA and YVAD-CMK or ABA abolished or attenuated the loss of RGCCs in the posterior retinas. CONCLUSIONS: NMDA-induced excitotoxicity involved apoptosis and caspases and PARP may play important roles in the pathways. PMID- 10476808 TI - Photoreceptor autophagy: effects of light history on number and opsin content of degradative vacuoles. AB - PURPOSE: To investigate whether regulation of rhodopsin levels as a response to changed lighting environment is performed by autophagic degradation of opsin in rod inner segments (RISs). METHODS: Groups of albino rats were kept in 3 lux or 200 lux. At 10 weeks of age, one group was transferred from 3 lux to 200 lux, another group was switched from 200 lux to 3 lux, and two groups remained in their native lighting (baselines). Rats were killed at days 1, 2, and 3 after switching. Another group was switched from 3 lux to 200 lux, and rats were killed at short intervals after the switch. Numbers of autophagic vacuoles (AVs) in RISs were counted, and immunogold labeling was performed for opsin and ubiquitin in electron microscopic sections. RESULTS: The number of AVs increased significantly after switching from 3 lux to 200 lux at days 1 and 2 and declined at day 3, whereas the reverse intensity change did not cause any increase. Early time points after change from 3 lux to 200 lux showed a significant increase of AVs 2 and 3 hours after switching. Distinct opsin label was observed in AVs of rats switched to 200 lux. Ubiquitin label was present in all investigated specimens and was also seen in AVs especially in 200-lux immigrants. CONCLUSIONS: Earlier studies had shown that an adjustment to new lighting environment is performed by changes in rhodopsin levels in ROSs. Autophagic degradation of opsin or rhodopsin may subserve, at least in part, the adaptation to abruptly increased habitat illuminance by removing surplus visual pigment. PMID- 10476809 TI - Remodeling of retinal capillaries in the diabetic hypertensive rat. AB - PURPOSE: To document the effect of sustained systemic hypertension on the integrity and ultrastructural morphology of retinal capillaries in diabetic and nondiabetic rats. METHODS: Normotensive (strain Wistar-Kyoto; WKY) and genetically hypertensive (spontaneously hypertensive; SHR) rats were rendered diabetic by intravenous streptozotocin injection. At 20 weeks of diabetes, endothelial cells, pericytes, and extracellular matrix were evaluated by ultrastructural morphometry. Serum albumin was localized by immunofluorescence microscopy. RESULTS: The endothelial cell layer was markedly thinner in the diabetic normotensive animals. The number of intercellular junctions was reduced in both the nondiabetic and diabetic hypertensive group but less so in the diabetic normotensive group. No significant endothelial cell loss was noted in either of the experimental groups, whereas the number of pericytes and the number of their cytoplasmic processes were reduced in diabetic and hypertensive animals. Significant thickening of the basement membrane and increased permeability to serum albumin were observed in diabetic and hypertensive rats and were strongly enhanced in the combined diseases. CONCLUSIONS: Endothelial thinning and shape changes from an elaborate to a simpler form as well as rounding up of the pericytes and loosening of their vascular sheaths indicate remodeling of the vascular wall during chronic diabetes and sustained hypertension, before a characteristic vasculopathy becomes manifest. The combination of diabetes and hypertension enhances these features, as well as basement membrane thickening and breakdown of the blood-retinal barrier. PMID- 10476810 TI - Development of VEP Vernier acuity and grating acuity in human infants. AB - PURPOSE: To compare the developmental sequences of two basic measures of pattern vision, Vernier acuity and grating acuity, using steady state visual-evoked potentials (VEPs) and an analysis designed to isolate pattern-specific responses from those due to motion in the Vernier stimulus. METHODS: The authors recorded VEPs from 57 healthy full-term infants and 4 adults. The grating acuity stimulus was a sinusoidal grating, temporally modulated (appearance-disappearance) at a rate of 3 Hz, with spatial frequency decreasing in linear steps during each 10 second trial. The Vernier acuity stimulus was a vertical square-wave grating with portions of each bar temporally modulated to make offsets appear and disappear at a rate of 3 Hz. Vernier offset size changed in log steps from small to large offsets. The authors recorded each observer's electroencephalogram (EEG) during multiple presentations of each stimulus type, and the EEG was digitized and filtered to obtain the amplitude and phase of the response at the first two harmonics of the stimulus temporal frequency. Thresholds were estimated with an extrapolation technique that took into account the signal-to-noise ratio and phase of the response. RESULTS: VEP Vernier acuity and grating acuity develop at different rates, with grating acuity approaching adult levels earlier than Vernier acuity. The within-subject relationship between VEP Vernier acuity and grating acuity follows the same developmental trajectory established by previous psychophysical studies of humans and monkeys. CONCLUSIONS: This VEP technique provides a rapid estimate of Vernier acuity in infants. VEP Vernier acuity remains strikingly immature throughout the first year of life, similar to behavioral Vernier acuity. Because Vernier acuity is a sensitive measure of amblyopia, this VEP test may be useful in the future to identify amblyopia and to follow its treatment progress in pediatric patients. PMID- 10476811 TI - Synthesis and release of docosahexaenoic acid by the RPE cells of prcd-affected dogs. AB - PURPOSE: Dogs affected with progressive rod-cone degeneration (prcd) have reduced levels of docosahexaenoic acid (DHA, 22:6n-3) in their plasma and rod photoreceptor outer segments (ROS). Dietary supplementation of DHA has failed to increase the ROS DHA levels to that of unaffected control dogs. The present study was undertaken to test the hypothesis that prcd-affected dogs have a reduced capacity for the synthesis and/or release of DHA in retinal pigment epithelial (RPE) cells. METHODS: RPE cells (first passage cultures) from prcd-affected and normal dogs were incubated with [3H]eicosapentaenoic acid (EPA, 20:5n-3) for 24 and 72 hours. After incubation, the radiolabeled fatty acids in the cells and media were analyzed. RESULTS: DHA and all its metabolic intermediates were detected in RPE cells from prcd-affected and normal dogs. No significant difference was found in the amount of products (including DHA) synthesized between normal and affected RPE cells at either time point. In the culture media, RPE cells from prcd-affected dogs released significantly more DHA than cells from normal dogs after 72-hour incubation, but not after 24-hour incubation. CONCLUSIONS: RPE cells from prcd-affected dogs can synthesize and release DHA at least as efficiently as cells from normal dogs. Therefore, synthesis of DHA from its precursor and its release from RPE cells does not appear to contribute to the reduction in ROS DHA levels found in prcd-affected animals. PMID- 10476812 TI - Increase in orthotopic murine corneal transplantation rejection rate with anterior synechiae. AB - PURPOSE: To evaluate the immunologic effect of anterior synechiae (AS) in a murine model of corneal transplantation. METHODS: Orthotopic penetrating keratoplasty with 12 interrupted sutures was performed on C57BL/6 donor mice and BALB/c recipient mice without AS (AS- group). In contrast to suturing in the AS- group, 3 of the 12 sutures were placed to create AS (AS+ group). The average graft opacity scores and rejection rates of both groups were compared. Cytotoxic T-lymphocyte (CTL) reactions and delayed hypersensitivity (DH) were evaluated 3 weeks after transplantation. Corneal cytokine expression was evaluated. RESULTS: The opacity scores of the AS+ group were consistently greater than those of the AS- group, and the rejection rate of the AS+ group was significantly greater than that of the AS- group (86% versus 54%, P = 0.03). The AS+ group had significantly higher CTL activity compared with the AS- group. There was no significant difference in DH between the two groups. The cytokine expression pattern in the AS+ group became similar to that of the AS- group in which the grafts were rejected. CONCLUSIONS: These findings indicate that AS impairs ocular immune privilege by mediating CTL activity, but without intensifying the DH response. Therefore, AS is a critical risk factor in allograft rejection in a murine model of corneal transplantation. PMID- 10476813 TI - Bioactivity of peptide analogs of the neutrophil chemoattractant, N-acetyl proline-glycine-proline. AB - PURPOSE: The release of N-acetyl-proline-glycine-proline (PGP), a chemoattractant resulting from direct alkaline hydrolysis of corneal proteins, is believed to be the initial trigger for neutrophil invasion into the alkali-injured cornea. The purpose of this study is twofold: (1) to compare the activity of N-acetyl-PGP with the bioactivities of other similar synthetic peptides in an effort to uncover information about this chemoattractant molecule, and (2) to test these peptide analogs as potential antagonists of N-acetyl-PGP. METHODS: The polarization assay was used to measure the potential chemotactic response of human neutrophils to peptides. Bioactivity was expressed as the peptide concentration required to produce 50% neutrophil polarization (EC50). Antagonist activity was expressed as the peptide concentration required to produce 50% inhibition (ID50) of polarization activated by N-acetyl-PGP. RESULTS: Peptide bioactivities (EC50) were ranked as follows: APGPR (0.34 mM) > N-acetyl-PGP (0.5 mM) > N-(PGP)4-PGLG (3 mM) = t-Boc-PGP (3 mM) > N-acetyl-PG (3.4 mM) > N-methyl PGP (15 mM) = PGP (15 mM) > peptides without detectable activity (t-Boc-PGP-OMe, N-acetyl-P, PG, PGG, GP, GG and gly-pro-hyp). Peptides with no detectable bioactivity were tested as potential antagonists of neutrophil polarization induced by N-acetyl-PGP. Gly-Pro-Hyp inhibited N-acetyl-PGP activation of polarization at 20 mM (ID50). No other synthetic peptide demonstrated a capacity for inhibition. CONCLUSIONS: The minimum requirement to elicit bioactivity was the presence of PGP alone or derivatives of PG in which the N-terminal proline is blocked. Using this approach, active and inactive mimetic peptides of N-acetyl PGP were produced. The most active peptide, APGPR, was equal to or slightly greater than N-acetyl-PGP, suggesting that more potent analogs might be designed. Gly-pro-hyp was the only inactive peptide analog to inhibit the chemoattractant. PMID- 10476814 TI - Effect of metalloproteinase inhibitor on corneal cytokine expression after alkali injury. AB - PURPOSE: Interleukin (IL)-1alpha and IL-6 levels in the cornea are greatly elevated during the early stages after an alkali burn in mice. The authors investigated the effect of synthetic inhibitor of matrix metalloproteinase (SIMP) on the expression of inflammatory cytokines in alkali-burned murine corneas and evaluated the clinical appearance of the eyes. METHODS: After 0.5N NaOH-alkali burns to 400 corneas of ICR mice, 200 received 400 microg/ml of SIMP topically 4 times a day while 200 corneas were similarly treated with vehicle only. At days 4, 7 and 14 after injury, each cornea was assigned a clinical score for corneal opacity, corneal epithelial defect, hyphema and cataract. Extracts of injured corneas in each group were then assayed for cytokine production using ELISA systems for IL-1alpha, IL-1beta, IL-6 and tumor necrosis factor-alpha (TNF alpha). RESULTS: The levels of IL-1alpha, IL-1beta and IL-6 were significantly lower in the SIMP-treated group than in the vehicle-treated group 7 days after the burn. However, levels of these cytokines were similar in the SIMP and non SIMP groups at days 4 and 14. Levels of TNF-alpha did not differ between both groups at any postinjury time. In the SIMP-treated corneas, there was less opacification and hyphema formation and epithelial regeneration was faster. CONCLUSIONS: Topical application of SIMP in alkali-burned murine corneas reduced the expression of IL-1alpha, IL-1beta, and IL-6 and lessened the severity of the injury. PMID- 10476815 TI - Asymmetric responses in cortical visually evoked potentials to motion are not derived from eye movements. AB - PURPOSE: Normal neonates and many adults after abnormal visual development have directional preferences for visual stimulus motions; i.e., they give better responses for optokinetic nystagmus (OKN) and visually evoked potentials (VEPs) in one direction than to those in the opposite direction. The authors tested whether the VEP responses were asymmetrical because of abnormal eye movements. METHODS: VEPs were recorded from the visual cortices of five macaque monkeys: one normal, one neonate, and three reared with alternating monocular occlusion (AMO). They were lightly anesthetized, followed by paralysis to prevent eye movements. They then had "jittered" vertical grating patterns presented in their visual fields. The steady state VEPs were analyzed with discrete Fourier transforms to obtain the amplitudes and phases of the asymmetries. RESULTS: The normal, control monkey had small, insignificant amplitudes of its asymmetrical Fourier component and random phases that were not 180 degrees out of phase across the left and right eyes. The neonatal monkey and the AMO monkeys all had large, significant asymmetries that were approximately 180 degrees out of phase between the left and right eyes. CONCLUSIONS: The neonate and abnormally reared monkeys continued to have asymmetrical responses even after their eyes were paralyzed. Therefore, eye movements cannot be the source of the asymmetrical amplitudes of the VEPs, and the visual cortex is at least one source responsible for asymmetries observed in neonates and adults reared under abnormal visual inputs. PMID- 10476816 TI - Elevation of human intraocular pressure at night under moderate illumination. AB - PURPOSE: An endogenous elevation of intraocular pressure (IOP) occurs at night in healthy young adults. The authors studied whether or not this IOP elevation can be detected under moderate illumination. METHODS: Twenty-five healthy volunteers, ages 18 to 25 years, were housed overnight in a sleep laboratory under a strictly controlled light-dark environment. Intraocular pressure was measured in the supine position every 2 hours, using a pneumatonometer. An 8-hour sleep period was assigned to each volunteer according to individual's accustomed sleep cycle. In the early part of this assigned period, sleep was encouraged with room lights off. Researchers performed IOP measurements at two time points with the aid of night vision goggles. In the middle to the late part of the assigned period, lights were turned on twice for a 1-hour interval. The light intensity was the same as before the bedtime. At the ending of each light period, IOP was measured under illumination. RESULTS: Average IOP was significantly higher in the assigned sleep period versus outside the period. The trough of mean IOP occurred just before the bedtime, and then IOP gradually increased and peaked at the end of the 8-hour assigned sleep period. The difference between the trough and peak IOP was 3.5 +/- 0.7 mm Hg (mean +/- SEM, n = 25). Within the assigned sleep period, the average IOP determined under illumination was significantly higher than the average IOP preceding the illumination. CONCLUSIONS: Elevation of IOP occurred during the assigned sleep period with two 1-hour light exposures of moderate intensity. Environmental light at night had no significant effect on the nocturnal IOP elevation in healthy young adults. PMID- 10476817 TI - Localization of cannabinoid CB1 receptors in the human anterior eye and retina. AB - PURPOSE: To determine the presence and distribution of CB1 cannabinoid receptors within the human eye. METHODS: A subtype-specific affinity-purified polyclonal antibody to the cannabinoid CB1 receptor was used to determine CB1 localization. Postmortem human eyes were fixed in methacarn and embedded in paraffin. Sagittal sections were mounted on slides and immunostained using antibodies to the CB1 receptor. Antibody binding was detected either by using peroxidase conjugated secondary antibodies and developing with diaminobenzidine or by using fluorescent secondary antibodies. RESULTS: Strong CB1 receptor labeling was detected in the ciliary epithelium, the corneal epithelium, and endothelium of the anterior human eye. Strong-to-moderate levels of CB1 staining were found in the trabecular meshwork and Schlemm's canal. Moderate labeling was detected in the ciliary muscle and in the blood vessels of the ciliary body. Moderate-to-light labeling also was detected in the sphincter papillae of the anterior human eye. Staining for CB1 receptors also was detected in human retina. The two synaptic layers of the retina and the inner and outer plexiform layers, were both moderately stained for CB1. In addition, moderate labeling was detected in the inner nuclear layer, and the ganglion cell layer. Strong labeling was detected in the outer segments of photoreceptors. No staining was observed in the corneal stroma or in the choroid. CONCLUSIONS: The wide distribution of cannabinoid CB1 receptors in both the anterior eye and the retina of humans suggests that cannabinoids influence several different physiological functions in the human eye. PMID- 10476819 TI - Vascular adhesion molecules in vitreous from eyes with proliferative diabetic retinopathy. AB - PURPOSE: To investigate whether proliferative vitreoretinopathy (PDR) is associated with a selective increase in vitreous levels of soluble vascular cell adhesion molecules that mediate leukocyte extravasation and interaction with endothelium during processes of inflammation and neovascularization. METHODS: Vitreous from 55 patients undergoing vitrectomy for treatment of PDR complicated by vitreous hemorrhage and/or traction retinal detachment was assayed for the presence of the soluble vascular cell adhesion molecules sICAM-1, sVCAM-1, and sE selectin using a standard enzyme-linked immunosorbent assays (ELISA). Vitreous from 12 cadaveric eyes matching age and sex of the patients were used as control samples. RESULTS: Vitreous levels of sICAM-1, sVCAM-1, and sE-selectin were significantly higher in eyes with PDR than in control cadaveric vitreous, and levels of all three molecules did not relate to the type or duration of diabetes mellitus. However, eyes with either traction retinal detachment alone or both traction retinal detachment and vitreous hemorrhage exhibited significantly higher levels of sICAM-1 and sE-selectin than eyes with vitreous hemorrhage alone. Vitreous levels of sVCAM-1 were similar in eyes with either vitreous hemorrhage or traction retinal detachment alone. CONCLUSIONS: The present observations suggest that molecular inflammatory mechanisms may contribute to processes of neovascularization and fibrosis observed in PDR, possibly not as the causative event, but as a result of endothelial, Muller, and retinal pigment epithelial cell activation. The results also indicate that retinal detachment amplifies the existing inflammation within the diabetic retina. Identification of any abnormalities in the production and control of specific adhesion molecules could have important implications in the design of new therapeutic regimens to treat and prevent this sight-threatening complication of diabetes mellitus. PMID- 10476818 TI - Regional differences in retinal vascular reactivity. AB - PURPOSE: Although glaucomatous visual field defects are more common in the superior field than in the inferior field, microaneurysms are more frequent in the superior than in the inferior retina in diabetic retinopathy. The authors hypothesized that differences in vascular hemodynamics in the two areas might contribute to these phenomena. METHODS: The blood flow response to hyperoxia and hypercapnia was evaluated in peripapillary retinal tissue superior and inferior to the optic nerve head using confocal scanning laser Doppler flowmetry. In 14 young, healthy persons, blood flow was measured while breathing room air and during isocapnic hyperoxia (100% O2 breathing) and isoxic hypercapnia (PCO2 increased 15% above baseline). Histograms were generated from pixel-by-pixel analysis of retinal portions of superior and inferior temporal quadrants of the entire image. RESULTS: Baseline blood flow in the inferior temporal quadrant was significantly greater than in the superior temporal quadrant (P < 0.05). However, the inferior region failed to increase in perfusion during hypercapnia and experienced significant mean blood flow reduction; flow reduction in the pixels at the 25th, 50th, 75th, and 90th percentile of flow; and an increased percentage of pixels without measurable flow, during hyperoxia (each P < 0.05). In contrast, in the superior temporal region, hyperoxia failed to reduce blood volume, velocity, or flow, whereas hypercapnia significantly increased mean flow; increased flow in the pixels at the 25th, 50th, 75th, and 90th percentile of flow; and reduced the percentage of pixels without measurable flow (each P < 0.05). CONCLUSIONS: The inferior temporal quadrant of the peripapillary retina is, in comparison with the superior temporas region, less responsive to vasodilation and more responsive to vasoconstriction. These differences could contribute to different susceptibility to visual field defect or vascular dysfunction in the superior and inferior retina. PMID- 10476820 TI - Acuity recovery and cone pigment regeneration after a bleach in patients with retinitis pigmentosa and rhodopsin mutations. AB - PURPOSE: To assess visual acuity recovery times and cone photopigment regeneration kinetics after a bleach in the fovea of patients with dominant retinitis pigmentosa due to rhodopsin mutations. METHODS: The authors measured acuity recovery times by computerized photostress testing in 13 patients with dominant retinitis pigmentosa and one of eight rhodopsin mutations. The authors also measured their time constants of cone photopigment regeneration with a video imaging fundus reflectometer to determine whether acuity recovery time depended on pigment regeneration kinetics. These values were compared with those of normal subjects, by the Mann-Whitney U test. The relationship between acuity recovery time and the time constant of cone photopigment regeneration among the patients was quantified by the Spearman rank correlation. RESULTS: The visual acuity recovery times, which averaged 22.0 seconds for the patients with retinitis pigmentosa and 11.2 seconds for the normal subjects, were significantly slower for the patient group (P < 0.001). The time constants of cone pigment regeneration, which averaged 172 seconds for the patients with retinitis pigmentosa and 118 seconds for the normal subjects, also were significantly slower for the patient group (P = 0.043). The authors also found a significant, positive correlation between the visual acuity recovery time and the time constant of pigment regeneration for the patients with retinitis pigmentosa (r = 0.65, P = 0.017). CONCLUSIONS: A slowing of foveal visual acuity recovery and cone pigment regeneration, which are related to each other, can occur in patients with retinitis pigmentosa, due to a rod-specific gene defect. PMID- 10476821 TI - Using interleukin 10 to interleukin 6 ratio to distinguish primary intraocular lymphoma and uveitis. PMID- 10476822 TI - Is lipofuscin eliminated from cells? PMID- 10476823 TI - A strange complex of diffuse congenital cardiovascular disease and cardiomyopathy, with localised myocardial calcification. AB - This unique case is of a man, followed clinically since infancy, who had a ventricular septal defect which closed spontaneously, a small arterial duct, and a minor degree of aortic coarctation, all without obvious symptoms. He later developed progressive cardiac failure which was attributed to some obscure form of congenital cardiomyopathy. He died at the age of 45 years. Necropsy showed a grossly abnormal arrangement of ventricular myocardial fascicles and bands, with absence of the papillary muscles causing tricuspid and mitral regurgitation. The various malformations are considered to be a gross example of a diffuse congenital cardiovascular disease complex, to the best of our knowledge previously undescribed. PMID- 10476824 TI - 'You didn't see them, but now you do!': use of balloon occlusion angiography in the identification of systemic venous anomalies before and after cavopulmonary procedures. AB - We describe the importance of angiographic identification of collateral venous channels by balloon occlusion venography after bidirectional cavopulmonary connections. Use of the balloon occlusion technique is essential for identification of these vessels, as they can easily be missed by standard venous angiography, with possible clinical consequences for postoperative management. PMID- 10476825 TI - Morphology of the elastic pulmonary arteries in pulmonary hypertension: a quantitative study. AB - Recent studies of intravascular ultrasound of the pulmonary arteries suggest that the technique can detect intimal and medial thickening in patients with pulmonary hypertension, potentially providing a method of assessing severity of pulmonary vascular disease in life. A major drawback of the technique is that only the elastic pulmonary arteries are accessible to current ultrasound catheters. The aim of this study was to determine whether morphological changes in vessels accessible to intravascular imaging reflect severity of pulmonary vascular disease and are of a sufficient degree to be detectable by current ultrasound catheters. Morphometric studies of the elastic pulmonary arteries were performed in specimens of lungs from 24 patients who had died with pulmonary hypertension (aged 3 weeks-9 years) and compared with measurements from infants who had died from sudden infant death syndrome. Morphological changes evident in the elastic pulmonary arteries in pulmonary hypertension included luminal dilation and medial thickening but these changes were too variable to be predictive of the severity of peripheral pulmonary vascular disease. Intimal thickening and atherosclerosis were present only in those with advanced pulmonary hypertensive disease. The changes of medial thickening and luminal dilation, nonetheless, are too variable to be predictive of the severity of peripheral pulmonary vascular disease. Thus, imaging by intravascular ultrasound may help confirm advanced pulmonary vascular disease, but is unlikely to differentiate less severe pulmonary vascular disease when the patient may still potentially be operable. PMID- 10476826 TI - Transcatheter embolization of congenital coronary arterial fistulas in adults. AB - In this report, we describe our experience with transcatheter occlusion of congenital coronary arterial fistulas in adults. From November 1992 to November 1996, 5 symptomatic patients, aged from 47 to 70 years, underwent transcatheter occlusion of fistulas using a retrograde arterial approach. All had chest pain or dyspnea on exertion. Detachable balloons were used in 4 patients, and Gianturco coils in 1. Detachable balloons were implanted through a Debrun system, while the coils were implanted through a 5 French right coronary Judkins catheter. Both were passed through an 8 French guiding catheter (Amplatz II). Each patient had a single fistula. The fistulas originated from the right coronary artery in 3 patients, and from the circumflex artery in 2. They drained into the pulmonary trunk in 3 patients, into the right atrium in 1, and into a bronchial artery in the other. All fistulas were occluded completely in the catheterization laboratory, and the procedures were uncomplicated. At follow up, 3 patients underwent coronary angiography, and there was no evidence of recanalization. Transcatheter embolization in adults of single congenital coronary fistulas with detachable balloons and coils is safe and effective and can be regarded as an acceptable alternative to surgery. PMID- 10476827 TI - Effect of radiofrequency catheter ablation of accessory pathways on autonomic tone in children. AB - BACKGROUND: Radiofrequency catheter ablation is standard treatment for children with re-entrant supraventricular tachycardias. Autonomic changes have been noted after such ablation for atrioventricular nodal re-entry tachycardia, but not as well documented with atrioventricular re-entry over an accessory pathway. METHODS AND RESULTS: In 10 normal paediatric volunteers and 12 children referred for electrophysiologic testing and radiofrequency ablation of supraventricular tachycardia, non-invasive autonomic function tests and tilt-table testing were performed, and the variability in 24-h heart rate was analysed. Patients with supraventricular tachycardia underwent these tests both 24-72 h before and 24 h after ablation. Patients with tachycardia underwent additional autonomic testing to assess the sensitivity of baroreceptors and the intrinsic heart rate with autonomic blockade immediately before and after ablation. One non-invasive autonomic function test, namely handgrip, demonstrated significant differences (p < 0.05) in diastolic blood pressure before and after ablation, though these values did not differ from controls. Significant decreases were noted in two indexes of the variability of heart rate before and after ablation (p < 0.05). Certain tilt test variables also demonstrated significant differences between controls and those with tachycardia subsequent to ablation. Intracardiac testing demonstrated changes (p < 0.05) in sinus cycle lengths, effective refractory periods and/or blood pressures at baseline and during testing of the sensitivity of baroreceptors before and after ablation. These changes were consistent with increased sympathetic or decreased parasympathetic tone. With autonomic blockade, these differences were abolished. CONCLUSIONS: Catheter ablation of accessory pathways in children was associated with changes consistent with increased sympathetic or decreased parasympathetic tone. These autonomic changes persisted 24 h after the ablation procedure. PMID- 10476828 TI - Right ventricular diastolic function after repair of tetralogy of Fallot: its relationship to the insertion of a 'transannular' patch. AB - Examined was the effect of surgical technique, particularly the insertion of a transannular patch, on right ventricular diastolic function, and the relationship of forward flow in the pulmonary arteries during late diastole to right ventricular diastolic function in patients with tetralogy of Fallot. Transtricuspid, superior caval venous and pulmonary arterial Doppler spectrals were obtained and compared between 44 patients who had been repaired with a transannular patch; 14 patients who had been repaired with muscular resection and/or pulmonary valvotomy; six who had been repaired with an infundubular patch; and 32 normal children. The velocities of forward flow during late diastole in the pulmonary arteries of normal children ranged from 19.8 to 29.4 cm s(-1) (mean 24.9 +/- 2.8 cm s(-1)) throughout the respiratory cycle. Restrictive right ventricular physiology, defined on the basis of increased forward flow in the pulmonary arteries during late diastole (> 30 cm s(-1)) was present in 25 (57%) of 44 patients with tetralogy of Fallot repaired using a transannular patch. Right ventricular volume was 50.1 +/- 23.7 cm3 in patients with a restrictive right ventricle and 64.9 +/- 21.4 cm3 in patients in whom the ventricle was non restrictive (p < 0.03). QRS duration was 140 +/- 18 and 156 +/- 24 ms in patients with restrictive and non-restrictive right ventricular physiology respectively (p < 0.003). Restrictive physiology was not encountered in patients with tetralogy in whom the pulmonary valve had been preserved. It is concluded that right ventricular restriction is present in many patients with tetralogy of Fallot at mid-term follow-up subsequent to repair using a 'transannular' patch. Restriction is associated with smaller right ventricular size and less prolongation of the QRS complex. PMID- 10476829 TI - Difficulties generated by the small, persistently patent, arterial duct. AB - Over recent years, echo-Doppler cardiography has shown that a small, sometimes silent, arterial duct exists in more patients than previously recognized. To know the incidence of an arterial duct subsequent to therapy, we studied retrospectively our patients undergoing open-heart surgery and surgical or catheter closure. Three groups of patients were studied: those with patency of the duct subsequent to open heart surgery without any sign of patency before or during surgery, those with persistent duct after surgical ligation and those with persistent patency after attempted catheter occlusion with the Rashkind device. In the first group (of 431 children) four (0.9%) had persistence of this duct, of which three were silent. In the second group, patency persisted in four of 100 patients (4%), three being silent. In the last group there were five persisting shunts, three producing no murmur, in 30 patients (17%). We compared our results with those reported in the literature and conclude that echo-Doppler cardiography is needed to detect persistent shunting across a duct after therapy, since most of the residual ducts in this study were silent. This means that clinical findings alone cannot be relied upon, and careful echo-Doppler cardiography is essential. Also, the process of closure of a persistent duct by surgical ligation or transcatheter intervention is no guarantee of success. The risk of infective endocarditis is important in such persistent ducts and, at present, it is unknown either for a small, silent duct or in a persistent duct that remains open after attempted transcatheter closure, but now is in association with a foreign body. PMID- 10476830 TI - Seasonality in live births with congenital heart disease in Malta. AB - This study was carried out to analyse seasonal variations in live births with congenital heart disease in the overall population of Malta. Included were all patients diagnosed as having congenital heart disease by echocardiography, cardiac catheterization, surgery or post mortem, by 1 year of age, and who were born between in Malta between 1990 and 1994. Results were analysed by X2 and by Edwards' cyclic method. There were 231 cases of live born congenital heart disease, which were divided into 114 cases not requiring intervention and 117 cases requiring intervention. Although the overall prevalence of congenital heart disease did not demonstrate any seasonal variation, the lesions requiring intervention showed a significant peak in September by Edwards' method (p = 0.03), which was enhanced by seasonal analysis (p = 0.003). These results were confirmed by conventional X2 and X2 for trend. The lesions not requiring intervention, which were comprised almost entirely (96%) of mild pulmonary stenosis and small ventricular septal defects, failed to show any seasonal trend. The literature regarding such seasonal trends in these malformations and statistical analysis of seasonality of congenital heart disease are reviewed. An environmental factor such as a maternal viral infection or treatment of such infections during the first trimester of pregnancy from November to January may precipitate congenital heart disease in predisposed Maltese foetuses. PMID- 10476831 TI - Relationship of the dimension of cardiac structures to body size: an echocardiographic study in normal infants and children. AB - Normalization of the dimensions of cardiac structures to the size of the body, using so-called Z scores, is becoming increasingly common in the management of infants and children with congenital heart disease. Current published nomograms for the ascertainment of Z scores for cardiac structures in childhood are based largely on normal data obtained in formalin-fixed hearts. Since decisions concerning management are frequently based on the findings of cross-sectional echocardiograms, the dimensions of 15 cardiac structures were measured using cross-sectional echocardiography in 125 normal infants and children. Regression equations were derived relating cardiac dimensions to the size of the body. The expression of size with the highest correlation to cardiac dimensions was body surface area. Nomograms were then developed from which the Z score of a cardiac structure could be estimated from a knowledge of the body surface area and the echocardiographically derived measurement. PMID- 10476832 TI - Jesse Efrem Edwards, MD. PMID- 10476833 TI - Double-outlet right ventricle with intact ventricular septum in a foetus with trisomy-18. AB - A rare case of double-outlet right ventricle with intact ventricular septum diagnosed by foetal echocardiography at 21 weeks of gestation is described. Amniocentesis revealed trisomy-18. The cardiac diagnosis was confirmed at autopsy. PMID- 10476834 TI - Divided left atrium associated with supravalvar mitral ring. AB - Reported is a case with a rare association of divided left atrium, supramitral stenosing ring of the left atrium, connection of the left superior caval vein to the roof of the left atrium, unroofed coronary sinus with an interatrial communication at the mouth of the unroofed sinus and ventricular septal defect. The need for a complete echocardiographic examination in the presence of pulmonary venous obstruction is emphasized. Surgery was successful in spite of significant preoperative pulmonary hypertension. PMID- 10476835 TI - Divided right atrium. Diagnosis by echocardiography, and considerations on the functional role of the Eustachian valve. AB - A child presented at birth with severe cyanosis. Echocardiography showed hypoplasia of the right heart with a right-to-left shunt at atrial level. A conservative approach was adopted initially, and the situation improved over a few months, with reversal of the atrial shunt. Surgery was successfully performed at 4 years of age after further echocardiography revealed a congenitally large Eustachian valve and an atrial septal defect. PMID- 10476836 TI - Sequential segmental approach to fetal congenital heart disease. AB - The sequential segmental approach is now universally used in the diagnosis of congenital heart disease. Its utilization during fetal sonography has not been well described. In this review, we show how the fetal heart can be approached in a sequential segmental manner by using six basic sonographic views. The transverse view of the fetal upper abdomen is obtained to determine the arrangement of the abdominal organs, which, in most cases, provides the important clues to the determination of the atrial arrangement. The four-chamber view is obtained to evaluate the atrioventricular junctions. The views of the left and right ventricular outflow tracts are obtained to evaluate the ventriculoarterial junctions. The three-vessel view and the aortic arch view are obtained for the evaluation of the arrangement and size of the great arteries, which provides the additional clues to the diagnosis of the abnormalities involving the ventriculoarterial junctions and the great arteries. The standard protocol to acquire these six basic views is also introduced. PMID- 10476837 TI - Intravascular ultrasonography of a patent arterial duct. PMID- 10476838 TI - Elastic modulus and hardness of cortical and trabecular bone lamellae measured by nanoindentation in the human femur. AB - The mechanical properties of bone tissue are determined by composition as well as structural, microstructural and nanostructural organization. The aim of this study was to quantify the elastic properties of bone at the lamellar level and compare these properties among osteonal, interstitial and trabecular microstructures from the diaphysis and the neck of the human femur. A nanoindentation technique with a custom irrigation system was used for simultaneously measuring force and displacement of a diamond tip pressed 500 nm into the moist bone tissue. An isotropic elastic modulus was calculated from the unloading curve with an assumed Poisson ratio of 0.3, while hardness was defined as the maximal force divided by the corresponding contact area. The elastic moduli ranged from 6.9 +/- 4.3 GPa in trabecular tissue from the femoral neck of a 74 yr old female up to 25.0 +/- 4.3 GPa in interstitial tissue from the diaphyseal cortex of a 69 yr old female. The mean elastic modulus was found to be significantly influenced by the type of lamella (p < 10(-6)) and by donor (p < 10(-6)). The interaction between the type of lamella and the donor was also highly significant (p < 10(-6)). Hardness followed a similar distribution as elastic modulus among types of lamellae and donor, but with lower statistical contrast. It is concluded that the nanostructure of bone tissue must differ substantially among lamellar types, anatomical sites and individuals and suggests that tissue heterogeneity is of potential importance in bone fragility and adaptation. PMID- 10476839 TI - Femoral strength is better predicted by finite element models than QCT and DXA. AB - Clinicians and patients would benefit if accurate methods of predicting and monitoring bone strength in-vivo were available. A group of 51 human femurs (age range 21-93; 23 females, 28 males) were evaluated for bone density and geometry using quantitative computed tomography (QCT) and dual energy X-ray absorptiometry (DXA). Regional bone density and dimensions obtained from QCT and DXA were used to develop statistical models to predict femoral strength ex vivo. The QCT data also formed the basis of a three-dimensional finite element (FE) models to predict structural stiffness. The femurs were separated into two groups; a model training set (n = 25) was used to develop statistical models to predict ultimate load, and a test set (n = 26) was used to validate these models. The main goal of this study was to test the ability of DXA, QCT and FE techniques to predict fracture load non-invasively, in a simple load configuration which produces predominantly femoral neck fractures. The load configuration simulated the single stance phase portion of normal gait; in 87% of the specimens, clinical appearing sub-capital fractures were produced. The training/test study design provided a tool to validate that the predictive models were reliable when used on specimens with "unknown" strength characteristics. The FE method explained at least 20% more of the variance in strength than the DXA models. Planned refinements of the FE technique are expected to further improve these results. Three-dimensional FE models are a promising method for predicting fracture load, and may be useful in monitoring strength changes in vivo. PMID- 10476840 TI - The association between negative muscle work and pedaling rate. AB - The objective of this research was to use a pedal force decomposition approach to quantify the amount of negative muscular crank torque generated by a group of competitive cyclists across a range of pedaling rates. We hypothesized that negative muscular crank torque increases at high pedaling rates as a result of the activation dynamics associated with muscle force development and the need for movement control, and that there is a correlation between negative muscular crank torque and pedaling rate. To test this hypothesis, data were collected during 60, 75, 90, 105 and 120 revolutions per minute (rpm) pedaling at a power output of 260 W. The statistical analysis supported our hypothesis. A significant pedaling rate effect was detected in the average negative muscular crank torque with all pedaling rates significantly different from each other (p < 0.05). There was no negative muscular crank torque generated at 60 rpm and negligible amounts at 75 and 90 rpm. But substantial negative muscular crank torque was generated at the two highest pedaling rates (105 and 120 rpm) that increased with increasing pedaling rates. This result suggested that there is a correlation between negative muscle work and the pedaling rates preferred by cyclists (near 90 rpm), and that the cyclists' ability to effectively accelerate the crank with the working muscles diminishes at high pedaling rates. PMID- 10476841 TI - Application of a fiber-reinforced continuum theory to multiple deformations of the annulus fibrosus. AB - Accurate tissue stress predictions for the annulus fibrosus are essential for understanding the factors that cause or contribute to disc degeneration and mechanical failure. Current computational models used to predict in vivo disc stresses utilize material laws for annular tissue that are not rigorously validated against experimental data. Consequently, predictions of disc stress resulting from physical activities may be inaccurate and therefore unreliable as a basis for defining mechanical-biologic injury criteria. To address this need we present a model for the annulus as an isotropic ground substance reinforced with two families of collagen fibers, and an approach for determining the material constants by simultaneous consideration of multiple experimental data sets. Two strain energy functions for the annulus are proposed and used in the theory to derive the constitutive equations relating the stress to pure stretch deformations. These equations are applied to four distinct experimental protocols and the material constants are determined from a simultaneous, nonlinear regression analysis. Good agreement between theory and experiment is achieved when the invariants are included within multiple, separate exponentials in the strain energy function. PMID- 10476842 TI - Contact analysis of biphasic transversely isotropic cartilage layers and correlations with tissue failure. AB - Failure of articular cartilage has been investigated experimentally and theoretically, but there is only partial agreement between observed failure and predicted regions of peak stresses. Since trauma and repetitive stress are implicated in the etiopathogenesis of osteoarthritis, it is important to develop cartilage models which correctly predict sites of high stresses. Cartilage is anisotropic and inhomogeneous, though it has been difficult to incorporate these complexities into engineering analyses. The objectives of this study are to demonstrate that a transversely isotropic, biphasic model of cartilage can provide agreement between predicted regions of high stresses and observed regions of cartilage failure and that with transverse isotropy cartilage stresses are more sensitive to convexity and concavity of the surfaces than with isotropy. These objectives are achieved by solving problems of diarthrodial joint contact by the finite-element method. Results demonstrate that transversely isotropic models predict peak stresses at the cartilage surface and the cartilage-bone interface, in agreement with sites of fissures following impact loading; isotropic models predict peak stresses only at the cartilage-bone interface. Also, when convex cartilage layers contacted concave layers in this study, the highest tensile stresses occur in the convex layer for transversely isotropic models; no such differences are found with isotropic models. The significance of this study is that it establishes a threshold of modeling complexity for articular cartilage that provides good agreement with experimental observations under impact loading and that surface curvatures significantly affect stress and strain within cartilage when using a biphasic transversely isotropic model. PMID- 10476843 TI - Evaluation of a testing method for the fatigue performance of total knee tibial trays. AB - Catastrophic failure of tibial baseplates due to fatigue fracture have occurred in patients. The International Standards Organisation (ISO) have proposed an endurance test to ensure a safety level for trays with respect to this problem. At present the magnitude of the applied load has not been defined. The purpose of this study is to evaluate the ISO test by comparing the results of two tibial trays: one with a clinical history of fracture of the tibial baseplate in some cases (Kinematic) and one without such a record (Kinemax). A load of 2000 N was selected, representing the peak force during the normal walking cycle. Finite element analysis (FEA) predicted that both the Kinematic and Kinemax trays would rapidly fail for two defined contact positions. Mechanical testing performed on both these components confirmed the FEA predictions. However, under service conditions, the Kinematic tray has been reported to have a failure of only 1-2%, the failure occurring at a mean of 54 months for early fracture and 7 years for late fracture. There are no published reports of failed Kinemax trays. It is concluded that the ISO test is an exaggeration of reality when a physiological load is used. As a result of our study we propose that a load of 500 N would effectively screen components with respect to the ISO test conditions for the likelihood of fracture. PMID- 10476844 TI - Lubrication of the human ankle joint in walking with the synovial fluid filtrated by the cartilage with the surface zone worn out: steady pure sliding motion. AB - A mixture model of synovial fluid filtration by cartilage in the human ankle joint during walking is presented for steady sliding motion of the articular surfaces. In the paper the cartilage surface zone is assumed worn out. The same model has been recently applied to the squeeze-film problem for the human hip joint loaded by the body weight during standing (Hlavacek, Journal of Biomechanics 26, 1145-1150, 1151-1160, 1993; Hlavacek and Novak, Journal of Biomechanics 28, 1193-1198, 1199-1205, 1995). The linear biphasic model for cartilage (elastic porous matrix + ideal fluid) due to Prof. V. C. Mow and his co workers and the biphasic model for synovial fluid (viscous fluid + ideal fluid), as used in the above-mentioned squeeze-film problem, are applied. For the physiologic parameters of the ankle joint during walking, a continuous synovial fluid film about 1 microm thick is maintained under steady entraining motion according to the classical model without the fluid transport across the articular surface. This is not the case in the filtration model with the cartilage surface zones worn out. On the contrary, this filtration model indicates that synovial fluid is intensively filtrated by such cartilage, so that no continuous fluid film is maintained and a synovial gel layer, about 10(-8) m thick, develops over the majority of the contact. Thus, if the cartilage surface zones are worn out, boundary lubrication should prevail in the ankle joint under steady sliding motion for the mean values of loading and the sliding velocity encountered in walking cycle. PMID- 10476845 TI - A fabric-dependent fracture criterion for bone. AB - A fracture criterion for bone tissue is proposed. Bone material is considered to be anisotropic and its properties are described by invoking the concept of directional variation of porosity. The fracture criterion is expressed as a scalar-valued function of the stress tensor and it incorporates an orientation dependent distribution of compressive/tensile strength. The proposed mathematical framework is applied to a numerical analysis of fracture in the proximal femur due to a fall from standing height. The risk of fracture is assessed in the context of two different porosity distributions, simulating a healthy and an osteoporotic bone. PMID- 10476846 TI - Experimental and numerical study of pulsatile flows through stenosis: wall shear stress analysis. AB - Different shapes of pulsatile flows through a model of stenosis are experimentally and numerically modeled to validate both methods and to determine the wall shear stress temporal evolution downstream from the stenosis. Two dimensional velocity measurements are performed in a 75% severity stenosis using a pulsed Doppler ultrasonic velocimeter. Finite element package is employed for the transient numerical simulations. Polynomial method, based on the experimental velocity values, is proposed to determine the wall shear stress temporal evolution. There is a good agreement between the numerical and experimental results. The wall shear stress temporal analysis shows oscillating wall shear stress values during the cycle with high wall shear stress values at the throat of about 120 dyn/cm2, and low values downstream from the stenosis of about - 2.5 dyn/cm2. The key result of the study is that the presence of the stenosis leads the artery to work in a direction which is opposite to the direction of a healthy artery. PMID- 10476847 TI - Glenoid cancellous bone strength and modulus. AB - The objectives of this study were to determine the strength and modulus of glenoid cancellous bone, including regional variations. The motivations were: to select a suitable bone substitute for standardized testing of glenoid prosthesis loosening, to assist in shoulder prosthesis design and to provide input data for finite element analyses. Ten glenoids from eight cadavers (mean age, 81) were tested by in situ indentation. Mean strength ranged from 6.7 to 17 MPa for the ten glenoids, the overall mean being 10.3 MPa. Mean E moduli ranged from 67 to 171 MPa for the individual glenoids, the overall mean being 99 MPa. These values are likely at the lower end of what would be expected for normal bone since strength and modulus decrease with age and the available specimens were older. These values may be appropriate for prosthesis design, however, since mechanical properties are reduced in rheumatoid arthritic bone. Regional trends were very similar for modulus and strength. The strongest region was postero-superior. The central column, correlating with the keel position in many glenoid components, was weaker than both the anterior and posterior regions but deeper. A large drop in strength and modulus below the subchondral layer emphasizes the importance of maintaining this layer during prosthetic replacement. PMID- 10476848 TI - Biomechanical influences on balance recovery by stepping. AB - Stepping represents a common means for balance recovery after a perturbation to upright posture. Yet little is known regarding the biomechanical factors which determine whether a step succeeds in preventing a fall. In the present study, we developed a simple pendulum-spring model of balance recovery by stepping, and used this to assess how step length and step contact time influence the effort (leg contact force) and feasibility of balance recovery by stepping. We then compared model predictions of step characteristics which minimize leg contact force to experimentally observed values over a range of perturbation strengths. At all perturbation levels, experimentally observed step execution times were higher than optimal, and step lengths were smaller than optimal. However, the predicted increase in leg contact force associated with these deviations was substantial only for large perturbations. Furthermore, increases in the strength of the perturbation caused subjects to take larger, quicker steps, which reduced their predicted leg contact force. We interpret these data to reflect young subjects' desire to minimize recovery effort, subject to neuromuscular constraints on step execution time and step length. Finally, our model predicts that successful balance recovery by stepping is governed by a coupling between step length, step execution time, and leg strength, so that the feasibility of balance recovery decreases unless declines in one capacity are offset by enhancements in the others. This suggests that one's risk for falls may be affected more by small but diffuse neuromuscular impairments than by larger impairment in a single motor capacity. PMID- 10476849 TI - A discussion on the threshold limit for hemolysis related to Reynolds shear stress. AB - Turbulence-related damage to blood is a major problem with the use of prosthetic devices, such as mechanical heart valves. An often-cited paper by Sallam and Hwang (1984). Biorheology 21, 783-797) quantified the threshold for hemolysis to be about 400 N m(-2), a value that has hitherto contributed to the evaluation of the potential dangerousness of a medical implantable device. We propose a discussion of the mentioned experiment, based on the application of stress analysis concepts to the original measurements: this is necessary to assess the peak turbulence shear stress value that could have been found in Sallam and Hwangs experiment, with a suitable orientation of the measurement axes. The result of our theoretical discussion is that the threshold value of 400 N m(-2) could probably be considerably underestimated: following this point of view, a 3 D stress analysis shows that the peak turbulence shear stress at the inception of hemolysis should be at least 600 N m(-2). This result, obtained on the basis of the study of RBCs' response to a turbulent environment, indicates that blood particles are probably more resistant to short-time shear stresses than it was thought. PMID- 10476850 TI - Hip endoprosthesis for in vivo measurement of joint force and temperature. AB - Friction between the prosthetic head and acetabular cup increases the temperature in hip implants during activities like walking. A hip endoprosthesis was instrumented with sensors to measure the joint contact forces and the temperature distribution along the entire length of the titanium implant. Sensors and two inductively powered telemetry units are placed inside the hip implant and hermetically sealed against body fluids. Each telemetry unit contains an integrated 8-channel telemetry chip and a radio frequency transmitter. Force, temperature and power supply data are transmitted at different frequencies by two antennas to an external twin receiver. The inductive power supply is controlled by a personal computer. Force and temperature are monitored in real time and all data are stored on a video tape together with the patient's images. This paper describes the design and accuracy of the instrumented implant and the principal function of the external system components. PMID- 10476851 TI - Limb locomotion--speed distribution analysis as a new method for stance phase detection. AB - The stance phase is used for the determination of many parameters in motion analysis. In this technical note the authors present a new kinematical method for determination of stance phase. From the high-speed video data, the speed distribution of the horizontal motion of the distal limb is calculated. The speed with the maximum occurrence within the motion cycle defines the stance phase, and this speed is used as threshold for beginning and end of the stance phase. In seven horses the results obtained with the presented method were compared to synchronous stance phase determination using a force plate integrated in a hard track. The mean difference between the results was 10.8 ms, equalling 1.44% of mean stance phase duration. As a test, the presented method was applied to a horse trotting on the treadmill, and to a human walking on concrete. This article describes an easy and safe method for stance phase determination in continuous kinematic data and proves the reliability of the method by comparing it to kinetic stance phase detection. This method may be applied in several species and all gaits, on the treadmill and on firm ground. PMID- 10476852 TI - Confined and unconfined stress relaxation of cartilage: appropriateness of a transversely isotropic analysis. AB - Previous studies have shown that stress relaxation behavior of calf ulnar growth plate and chondroepiphysis cartilage can be described by a linear transverse isotropic biphasic model. The model provides a good fit to the observed unconfined compression transients when the out-of-plane Poisson's ratio is set to zero. This assumption is based on the observation that the equilibrium stress in the axial direction (deltaz) is the same in confined and unconfined compression, which implies that the radial stress deltar = 0 in confined compression. In our study, we further investigated the ability of the transversely isotropic model to describe confined and unconfined stress relaxation behavior of calf cartilage. A series of confined and unconfined stress relaxation tests were performed on calf articular cartilage (4.5 mm diameter, approximately 3.3 mm height) in a displacement-controlled compression apparatus capable of measuring delta(z) and delta(r). In equilibrium, delta(r) > 0 and delta(z) in confined compression was greater than in unconfined compression. Transient data at each strain were fitted by the linear transversely isotropic biphasic model and the material parameters were estimated. Although the model could provide good fits to the unconfined transients, the estimated parameters overpredicted the measured delta(r). Conversely, if the model was constrained to match equilibrium delta(r), the fits were poor. These findings suggest that the linear transversely isotropic biphasic model could not simultaneously describe the observed stress relaxation and equilibrium behavior of calf cartilage. PMID- 10476853 TI - A new friction tester of the flexor tendon. AB - We have developed a new device to measure the friction force and calculate the friction coefficient between a rabbit flexor tendon, a pulley and a proximal phalanx. The flexor digitorum fibularis tendon of a rabbit was taken intact with the proximal phalanx, and tendon pulleys were attached to both ends of the bone. Both ends of the tendon were clamped to acrylic plates and connected to stainless steel plates equipped with strain gauges. A pretension of 1.96 N was applied so as not to loosen the tendon. The proximal phalanx was fixed to an acrylic plate on the actuator, which gave 8 mm of transfer to the acrylic plate at a speed of 2 mm/s. The interface between the tendon and the surrounded tissue created the friction force, when the load was applied on the distal pulley. The friction force could be obtained from the difference between the tension of both ends of the tendon, which was measured with strain gauges and sampled with a personal computer. The friction force and the friction coefficient were calculated from the measured force and the applied load. The load and the pre-loading time, which was defined as loading duration before gliding, were varied in order to observe the change of the friction coefficient. The friction coefficient was not affected by the load and increased with the pre-loading time. The value of mu(s) ranged from 0.027 to 0.111 (0.072 +/- 0.023), and that of (mu)d ranged from 0.010 to 0.069 (0.039 +/- 0.014) (pre-loading time was 5 s). Our method will allow for the examination of various surgical treatments and lubricants. Moreover, it can be applied to other tissues of any animals with similar structures to the rabbit's digitorum. PMID- 10476854 TI - In regards to the "ISB recommendations for standardization in the reporting of kinematic data". PMID- 10476855 TI - A short and efficient synthesis of 4-[2,2-dimethyl-4(tol-4-yl)benzochrom-3-en-7 yl]benzoic acid - a potent retinoic acid receptor antagonist. AB - An efficient synthesis of a potent RAR antagonist is described starting from disubstituted beta-naphthol. The functional groups on 2 and 3 positions of the beta-naphthol 5 were elaborated into benzochromanone 8. The title compound was prepared by Suzuki coupling of the left and right hand pieces. PMID- 10476856 TI - A rapid method to identify exo-protease inhibitors. AB - A new approach to the evaluation of exo-protease inhibitor candidates is presented. The application of new water-soluble substrates that release organic soluble fluorescent groups upon proteolytic cleavage allows amplification of the assay signal via concentration of the cleavage product. A combinatorial library of disubstituted xanthenes designed to resemble a known inhibitor was screened and a new HLE inhibitor (Ki = 79 microM) was identified. PMID- 10476857 TI - Rational herbicide design by inhibition of tryptophan biosynthesis. AB - Compounds designed to mimic the tryptophan synthase alpha subunit reactive intermediate were found to be potent inhibitors of the enzyme. These compounds are herbicidal and the herbicidal mode of action was demonstrated to be due to disruption of tryptophan biosynthesis. PMID- 10476859 TI - Synthesis and electrochemical studies of a new iron tetra-catecholamide complex. AB - A new tetra-catecholamide compound N5,N6-thiodipropanoyl-bis[N1,N10-bis(2,3 dihydroxybenzoyl-spermidi ne)] (H8L) has been synthesised as an iron chelator of Fe (III). Cyclic voltammogram of the iron complex H2LFe run under an argon atmosphere shows a quasi-reversible redox process with E0 = -430 mV vs. SCE in CH3OH/H20 (60/40). This value approaches the range of biological reductants and consequently the complex may mimic the release of iron from enterobactin to the agents which are directly involved in cell metabolism. PMID- 10476858 TI - Synthesis and pharmacological activity of diarylindole derivatives. Cytotoxic agents based on combretastatins. AB - Taking into account the structure of Combretastatins, we have synthesized and assayed for cytotoxic activity of new indole derivatives. Two aryl groups are maintained in the cis orientation required for activity by means of an indole moiety built up on less active ketoderivatives used as starting materials. PMID- 10476860 TI - Electrochemical and esr spin trap studies of a new iron tetra-catecholamide complex. AB - A new siderophore, N5,N6-thiodipropanoyl-bis[N1,N10-bis(2,3-dihydroxy benzoyl spermidine)]-Fe (III) complex or H2LFe has been synthesised. The reaction of the reduced form of this complex with dioxygen has been investigated through electrochemical study and revealed the formation of a new species assumed to be H2O2. This species has been confirmed by esr spectroscopy using the diamagnetic compound 5-deutero-2,2,5-trimethylpyrrolidine-1-hydroxyl as spin trap. The resulting persistent radical is 5-deutero-2,2,5-trimethylpyrrolidine-1-yloxy (aN = 16.58 G, aD = 3.49 G). PMID- 10476861 TI - Synthesis and cytotoxic activity of N-(2-pyridylsulfenyl)urea derivatives. A new class of potential antineoplastic agents. AB - Starting from a 3D-model for the antineoplastic activity of diarylsulfonylureas several new features were proposed and tested. Both types of assayed compounds, the N-(2-pyridylsulfonyl)urea and N-(2-pyridylsulfenyl)urea derivatives, inhibited by 50% the growth of the CCRF-CEM cell line at a dosage near to 1 microM. The N -(2-pyrimidinyl) derivative of the sulfenylurea 6c showed a better profile against HT-29, K-562 and HTB-54 tumor cell lines than the corresponding sulfonylurea 6b. Structural modifications on aryl systems affected differently to the cytotoxic activity shown by the compounds against each cell line. PMID- 10476862 TI - Synthesis and antitumoral activities of marine ent-chromazonarol and related compounds. AB - Efficient syntheses of ent-isozonarol (6a), ent-isozonarone (7a) and ent chromazonarol (8) from (-)-sclareol (12) are described. 6a and 7a show a significative antitumoral activity. PMID- 10476863 TI - Synthesis and biological evaluation of conformationally restricted Gabapentin analogues. AB - A series of conformationally restricted Gabapentin analogues has been synthesised. The pyrrolidine analogue (R)-2-Aza-spiro[4.5]decane-4-carboxylic acid hydrochloride (3a) had an IC50 of 120 nM, similar to that of Gabapentin (IC50 = 140 nM), at the Gabapentin binding site on the alpha2delta subunit of a calcium channel. Compound (3a) also reversed carrageenan induced hyperalgesia in rats. PMID- 10476864 TI - Potent, easily synthesized huperzine A-tacrine hybrid acetylcholinesterase inhibitors. AB - Hybrid acetylcholinesterase inhibitors composed of a key fragment of huperzine A and an intact tacrine unit were prepared. The syntheses are quite direct, proceeding in a maximum of 4 linear steps from commercially available starting materials. The optimum hybrid inhibitor (+/-)-9g is 13-fold more potent than (-) huperzine A, and 25-fold more potent than tacrine. PMID- 10476865 TI - Synthesis of new (benzimidazolyl)piperazines with affinity for the 5-HT1A receptor via Pd(0) amination of bromobenzimidazoles. AB - The synthesis of a new family of (benzimidazolyl)piperazines has been developed through Pd(0) mediated amination of 4- and 6-bromobenzimidazole derivatives. Preliminary studies showed that some of these compounds are potent 5-HT1A receptor ligands. PMID- 10476866 TI - 8-acenaphthen-1-yl-1-phenyl-1,3,8-triaza-spiro[4.5]decan-4-one derivatives as orphanin FQ receptor agonists. AB - A series of 8-acenaphthen-1-yl-1-phenyl-1,3,8-triaza-spiro[4.5]decan+ ++-4-one derivatives 1 was studied with respect to the binding affinity for the orphanin FQ (OFQ) and opioid (mu, kappa, delta) receptors. The influence of stereochemistry as well as the substitution pattern of the phenyl-ring in position 1 on the affinity for the orphanin FQ receptor and selectivity to opioid (mu, kappa, delta) receptors is discussed. The most interesting compound 1c was tested for its anxiolytic-like properties in vivo. PMID- 10476867 TI - Synthesis and dopamine receptor modulating activity of unsubstituted and substituted triproline analogues of L-prolyl-L-leucyl-glycinamide (PLG). AB - Triprolines Pro-Pro-Pro-NH2 (4), Pro-Pro-D-Pro-NH2 (5), Pro-Pro(trans-3-Me)-D-Pro NH2 (6), and Pro-Pro(cis-3-Me)-D-Pro-NH2 (7) were made as conformationally constrained analogues of Pro-Leu-Gly-NH2. Triprolines 4-6 produced significant increases in the high- and low-affinity state ratio (RH/RL) of the dopamine receptor, but only 4 was found to increase apomorphine induced rotations in 6 hydroxydopamine-lesioned rats. PMID- 10476868 TI - Structure-based design and synthesis of a novel class of Src SH2 inhibitors. AB - The structure-based design and synthesis of a novel class of 2,4-disubstituted thiazoles as Src SH2 inhibitors is described. Initial results are presented, including the X-ray and NMR analysis of one thiazole inhibitor bound to Lck and Src SH2. PMID- 10476869 TI - Structure-activity relationships of a novel class of Src SH2 inhibitors. AB - The structure-activity relationships (SAR) of a novel class of Src SH2 inhibitors are described. Variation at the pY+1 and pY+3 side chain positions using 2,4- and 2,5-substituted thiazoles and 1,2,4-oxadiazoles as scaffolds resulted in inhibitors that bound as well as the standard tetrapeptide Ac-pYEEI-NH2. PMID- 10476870 TI - Hydroxyoxazolidines as alpha-aminoacetaldehye equivalents: novel inhibitors of calpain. AB - The synthesis of [1-[(5-hydroxy-4-(phenylmethyl)-3-oxazolidinyl)carbonyl]-2 ethylpropy lcarbamic acid phenylmethyl ester (2; MDL 104,903), a potent inhibitor of calpain, is described. Synthesis of related compounds, which offer insights into the mechanism of action for 2, are also described, as is an O-acetyl prodrug derivative of 2. PMID- 10476871 TI - P2-achiral, P'-extended alpha-ketoamide inhibitors of calpain I. AB - A series of potent P2-achiral, P'-extended alpha-ketoamide inhibitors of calpain I is described. PMID- 10476872 TI - Dopamine-selective response in membrane potential by homooxacalix[3]arene triether host incorporated in PVC liquid membrane. AB - Selectivities of membrane potential changes for catecholamines and inorganic cations were investigated with lipophilic derivatives of calix[6]arene and related hosts incorporated in poly(vinyl chloride) (PVC) matrix liquid membranes. Homooxacalix[3]arene triether displayed an excellent selectivity for dopamine against other catecholamines (adrenaline, noradrenaline) and also against inorganic cations (K+, Na+). PMID- 10476873 TI - Synthesis and estrogen receptor binding affinity of a porphyrin-estradiol conjugate for targeted photodynamic therapy of cancer. AB - A tetraphenylporphyrin-C11-beta-estradiol conjugate has been synthesized. Competitive binding assay of the conjugate with estrogen receptor (ER)-ligand binding domain showed that the conjugate binds specifically to the protein with high affinity. Potential use of this conjugate to selectively deliver cytotoxic porphyrins to ER-positive cells in various carcinomas is discussed. PMID- 10476874 TI - Synthesis and biological evaluation of novel 1beta-methylcarbapenems having a new moiety at C-2. AB - The synthesis and biological activity of the novel series of 1 beta methylcarbapenems 1a-f, bearing a variety of 3",4"-disubstituted pyrrolidinamides as substituents at C-2, are described. Of these carbapenems, diol 1a showed the most potent and well balanced antibacterial activity against Gram-positive and Gram-negative. 1a was also evaluated for pharmacokinetics and in vivo therapeutic efficacy in systemic infections. PMID- 10476876 TI - Molecular structure of FR901277, a novel inhibitor of human leukocyte elastase, and its binding mode simulation. AB - X-ray crystal structure analysis of FR901277, a novel inhibitor of human leukocyte elastase, was performed and revealed that the lipophilic side chains are located towards the outside of the molecule. Binding simulation using computational methods showed that these lipophilic moieties could bind to the hydrophobic binding pockets of HLE. PMID- 10476875 TI - Substituted indoles as potent and orally active 5-lipoxygenase activating protein (FLAP) inhibitors. AB - This paper reports on the SAR investigation of inhibitors of 5-lipoxygenase activating protein (FLAP) based on MK-0591. Emphasis was made on modifications to the nature of the link between the indole and the quinoline moieties, to the substitution pattern around the two heterocycles and to possible replacements of the quinoline moiety. Lead optimization culminated in (3-[1-(4-chlorobenzyl)-3-(t butylthio)-5-(pyridin-2-ylmethoxy)-ind ol-2-yl]-2,2-dimethylpropanoic acid (18k), as a potent inhibitor of leukotriene biosynthesis that is well absorbed and active in functional models. PMID- 10476877 TI - NMR-based discovery of phosphotyrosine mimetics that bind to the Lck SH2 domain. AB - Using an NMR-based screen, a series of novel phosphotyrosine mimetics were discovered that bind to the SH2 domain of Lck. These compounds may serve as useful leads for the design of nonpeptide inhibitors of SH2 domains with improved bioavailability and metabolic stability compared to the natural ligands that contain phosphotyrosine. PMID- 10476878 TI - Naphthazarin derivatives (II): formation of glutathione conjugate, inhibition of DNA topoisomerase-I and cytotoxicity. AB - 6-(1-Hydroxyalkyl)-5,8-dimethoxy-1,4-naphthoquinones, expressing a higher reactivity in conjugation with glutathione, showed a greater potency in the inhibition of DNA topoisomerase-I and the cytotoxicity against L1210 cells than 2 (1-hydroxyalkyl)-DMNQ derivatives, implying the participation of electrophilic arylation in the bioactivities. In further study 6-(1-Hydroxyalkyl)-5,8-dimethoxy 1,4-naphthoquinones with an alkyl group of shorter chain length (C2-C6) exerted a greater bioactivities than those with longer chain length(>C6). PMID- 10476879 TI - Selective A1-adenosine receptor antagonists identified using yeast Saccharomyces cerevisiae functional assays. AB - Evaluation of a biased "library" of pyrrolo[2,3-d]pyrimidines using yeast-based functional assays expressing human A1- and A2a-adenosine receptors, led to the A1 selective antagonist 4b. A direct correlation between yeast functional activity and binding data was established. Practical compounds with polar residues at C-4 of the pyrrolopyrimidine system required H-bond donor functionality for high potency. PMID- 10476880 TI - Opsin shift in an aldolase antibody. AB - An antibody-retinal assembly that mimics the opsin shift (OS) of the naturally occurring visual pigments is reported. Both experiments and calculations show that the aldolase antibody 33F12 covalently binds all-trans retinal via a protonated Schiff base with a lysine residue. This chromophore, which exhibits a remarkable opsin red shift (140 nm), represents a useful model system for studying the factors that contribute to the OS. PMID- 10476881 TI - 8-aminoquinolines as anticoccidials - Part III. AB - Analogues of the antimalarial pentaquine, 1, in which the nature of the side chain on the 8-amino position was varied, were prepared and evaluated for anticoccidial activity both in vitro and in vivo. Specifically, both the inter nitrogen distance and the nature of the terminal amino group were investigated. Novel analogues of equal or improved efficacy in vitro and in vivo to pentaquine were discovered. PMID- 10476882 TI - Probing the role of interfacial residues in a dimerization inhibitor of HIV-1 protease. AB - The importance of each side chain of a cross-linked interfacial peptide inhibitor of HIV-1 protease was evaluated using an alanine scanning approach. Whereas the parent inhibitor has an IC50 value of 350 nM, values for the mutations reported here range from 280-9200 nM. The relative importance or each residue was thus assigned and correlated to the solvent accessible surface area (SASA) exposed upon mutation. PMID- 10476883 TI - Investigations into self-association of vancomycin covalent dimers using surface plasmon resonance technology. AB - Covalent dimers of vancomycin linked through the vancosamine sugar moieties of the glycopeptide antibiotic have been synthesized in one step in 67-69% yield. The propensity for self-association of these and related vancomycin covalent dimers is evaluated using surface plasmon resonance technology. PMID- 10476884 TI - Synthesis of a phostone glycomimetic of the endothelin converting enzyme inhibitor phosphoramidon. AB - The phostone analog of phosphoramidon, an inhibitor of endothelin converting enzyme, was synthesized from L-rhamnose. Coupling of the cyclic phosphonic acid with the dipeptide H-Leu-Trp-OMe gave, after deprotection and purification by reverse-phase HPLC, the desired phostone which exhibited an IC50 of 5.05+/-2.7 microM. PMID- 10476885 TI - Synthesis and biological activity of substituted bis-(4-hydroxyphenyl)methanes as N-type calcium channel blockers. AB - Voltage activated calcium channel (VACC) blockers have been demonstrated to have utility in the treatment of stroke and pain. A series of aminomethyl substituted phenol derivatives has been identified with good functional activity and selectivity for N-type VACC's over sodium and potassium channels. The methods of synthesis and preliminary pharmacology are discussed herein. PMID- 10476886 TI - Structure-activity relationship at the proximal phenyl group in a series of non peptidyl N-type calcium channel antagonists. AB - Selective N-Type Voltage Sensitive Calcium Channel (VSCC) antagonists have shown utility in several models of pain and ischemia. We report the structure-activity relationship at the proximal phenyl group in a series of non-peptidyl VSCC blockers. PMID- 10476887 TI - Influence of estrogen and osteopenia/osteoporosis on clinical periodontitis in postmenopausal women. AB - BACKGROUND: In Western societies, more than one-third of the female population above age 65 suffers from signs and symptoms of osteoporosis, a disorder characterized by low bone mass. Estrogen deficiency is the dominant pathogenic factor for osteoporosis in women. The impact of estrogen deficiency and osteopenia/osteoporosis on periodontitis is unclear, partially due to the lack of longitudinal studies evaluating clinical signs of gingival inflammation and periodontitis progression. The purpose of this investigation was to analyze prospectively the influence of serum estradiol levels and osteopenia/osteoporosis on common clinical measurements of periodontal disease over a 2-year period. METHODS: Fifty-nine moderate/advanced adult periodontitis patients and 16 non periodontitis subjects, all within 5 years after menopause at baseline, completed the study. Serum estradiol levels (E2) were measured yearly by 125I radioimmunoassay, and osteopenia/osteoporosis was determined by dual energy x-ray absorptiometry of the lumbar spine. Posterior interproximal clinical measurements were obtained every 6 months for the periodontitis patients, including explorer detectable supragingival plaque, bleeding on probing (BOP) and relative clinical attachment level (RCAL). Baseline probing depths, smoking history, and demographic data also were collected. RESULTS: Data indicated that baseline demographic measurements and bone mineral density (BMD) of the lumbar spine were not different between E2-deficient and E2-sufficient subjects. Smoking activity (packs smoked/day, years smoked) was higher in periodontitis patients (P=0.0001). E2-sufficient periodontitis subjects had a higher frequency of supragingival plaque without increasing gingival inflammation. E2 status did not influence the percentage of sites losing RCAL for either periodontitis or non-periodontitis groups, but when non-smoking osteopenic/osteoporotic periodontitis patients were evaluated, E2-deficient subjects had more BOP (43.8% versus 24.4%, P<0.04) and a trend toward a higher frequency of > or =2.0 mm RCAL loss (3.8% versus 1.2%, P<0.1) than E2-sufficient subjects. CONCLUSIONS: These data suggest that E2 supplementation (serum E2>40 pg/ml) is associated with reduced gingival inflammation and a reduced frequency of clinical attachment loss in osteopenic/osteoporotic women in early menopause. PMID- 10476889 TI - Modification of an osteoconductive anorganic bovine bone mineral matrix with growth factors. AB - BACKGROUND: Osteoconductive anorganic bovine bone mineral matrix material has been used clinically in bone regeneration procedures. Platelet-derived growth factor-BB (PDGF-BB) and insulin-like growth factor (IGF-I) are important anabolic growth factors for bone. It was the aim of these studies to 1) examine the interaction of this bone graft material with PDGF-BB and IGF-I and 2) determine if the combination of growth factors with the matrix could stimulate osteoblastic cell proliferation. METHODS: Adsorption of PDGF-BB and IGF-I was done using 125I radio-labeled growth factors. The PDGF-BB or IGF-I was incubated with the anorganic bovine bone matrix, and the amount of adsorbed growth factor was measured. In the desorption studies, radiolabeled growth factors were adsorbed to the matrix material. The samples were incubated in buffer for various time periods, and the amount remaining on the matrix was measured to calculate the percentage of released growth factor. The biological activity was tested in an in vitro assay with primary culture neonatal rat osteoblastic cells. Porous bone matrix with known amounts of adsorbed PDGF-BB or IGF-I was produced. The osteoblastic cells were cultured on the bone mineral matrix, with and without adsorbed growth factor, and proliferation was assessed by 3H-thymidine incorporation. RESULTS: Both PDGF-BB and IGF-I adsorbed to bone mineral matrix in a concentration-dependent fashion. The affinity of IGF-I for the material was 10 fold greater than PDGF-BB. In the experiments that measured the release of the initially adsorbed growth factors, approximately 50% of the PDGF-BB and 10% of the IGF-I were released after 10 days. PDGF-BB adsorbed to the matrix material significantly (P <0.05, ANOVA) enhanced the proliferation of cultured osteoblastic cells compared to the mineralized matrix alone. However, IGF-I adsorbed to the matrix material did not significantly enhance cell proliferation. CONCLUSIONS: These results suggest that PDGF-BB can be adsorbed to the anorganic bovine bone mineral matrix and that this growth factor subsequently enhances the osteogenic properties of this bone graft material. IGF-I also adsorbed to the graft material; however, it was not readily released and it did not produce significant effects in the biologic assay. It appears that it may be clinically feasible to adsorb PDGF to anorganic bovine bone and that this combination of bone growth factor and mineral matrix has the potential for clinical applications. PMID- 10476888 TI - Relationships between longitudinal changes in radiographic alveolar bone height and probing depth measurements: data from postmenopausal women. AB - BACKGROUND: The relationship between loss of radiographic alveolar bone height and probing attachment loss has been studied by a number of investigators, with mixed results. Recent studies have found weak correlations and have suggested that the relationship between bone loss and attachment loss is complex, perhaps because changes in bone height and attachment level are separated in time. METHODS: The 85 patients in this report were part of a prospective estrogen replacement interventional study. All patients were in good oral health at entry and received annual oral prophylaxis as part of the study. Standard probing measurements were made with a pressure-sensitive probe at 6 sites on each tooth. Vertical bite-wing radiographs were taken of each patient, radiographs were digitized, and 6 linear measurements (corresponding to probing site measurements) were made from the cemento-enamel junction to the alveolar crest. These procedures were performed at baseline and at annual intervals; this study reports results after 2 years. Data were analyzed both by individual site and by averaging identical sites from all measured teeth for each patient. RESULTS: Very weak direct relationships between change in alveolar bone height and change in attachment level were found in both the site data (r2=0.0022; P = 0.189) and the patient average data (r2=0.031; P= 0.104). CONCLUSIONS: The changes in these patients were probably due to systemic changes in bone health rather than to periodontal disease. However, the weak correlations between changes in attachment level and bone height are similar to recent studies of periodontal disease. Our results support suggestions in the literature that the link between changes in attachment and alveolar bone height is complex, perhaps because changes in the 2 tissue types are separated by a considerable time delay. PMID- 10476890 TI - A comparison of the efficacy of 2 powered toothbrushes in affecting plaque accumulation, gingivitis, and gingival bleeding. AB - BACKGROUND: The purpose of this 30-day blinded, parallel-design study was to compare the effect of 2 powered toothbrushes, the Rowenta MH700 and the Braun Plak Control Ultra, on reducing plaque accumulation, gingivitis, and gingival bleeding in a cohort of 60 healthy adults. METHODS: After baseline evaluation of plaque, gingivitis, and gingival bleeding, subjects were randomly assigned to one of the experimental groups, provided written and verbal toothbrushing instructions, and had their teeth polished. At 2 weeks (follow-up 1) and 4 weeks (follow-up 2), all clinical parameters were again evaluated. RESULTS: The Braun group demonstrated a nearly significant reduction in gingival index (GI) and a significant reduction in GI at follow-up 2. The Rowenta group demonstrated significant reductions in GI, plaque index (PI), and bleeding index (GBI) at both follow-up 1 and 2 examinations. At follow-up 1, the Braun group demonstrated a nearly significant reduction in GI, a significant reduction in PI, and a non significant reduction in GBI. At follow-up 2, the Braun group demonstrated a significant reduction in GBI, but not a significant reduction in PI. The reduction in GI for the Rowenta group was significantly greater (P values of 0.0001 and 0.0001, respectively) than that demonstrated in the Braun group. However, the Rowenta group did not demonstrate a significantly greater reduction in PI (P values of 0.7135 and 0.3184 for follow-up 1 and follow-up 2, respectively) or GBI than the Braun group at either examination (P values of 0.0663 and 0.3397 for follow-up 1 and 2, respectively). CONCLUSIONS: The results of this study support the findings of numerous other studies that powered toothbrushes have great potential to remove plaque and improve gingival health and that the improvement can be demonstrated in a relatively short period of time. PMID- 10476891 TI - Vascular endothelial growth factors and progression of periodontal diseases. AB - BACKGROUND: Tissues become hemorrhagic and edematous coincident to periodontal diseases; however, there is little information concerning the biologic mechanisms which may produce these changes. Vascular endothelial growth factor (VEGF) is a macromolecule which enhances blood vessel growth and permeability. However, there is no information concerning gingival VEGF concentrations within normal or diseased gingiva. The purpose of this study was to assess changes in gingival concentrations of VEGF during initiation and progression of periodontal diseases and compare them to changes in the number of blood vessel profiles and concentration of recognized markers of periodontal disease severity (interleukin 6[IL-6]). METHODS: Normal (non-hemorrhagic gingiva adjacent to a < or =3 mm gingival sulcus) and inflamed gingiva (hemorrhagic gingiva adjacent to a < or =3 mm, 4 to 6 mm, or >6 mm periodontal pocket) were studied. VEGF and IL-6 concentrations were assessed by ELISA and the number of blood vessels determined by histomorphometric techniques. Data were placed into one of the following groups: < or =3 mm, normal; < or =3 mm, diseased; 4 to 6 mm, diseased; and >6 mm, diseased. These groups were compared by factorial ANOVA and Scheffe comparisons. In addition, groups were compared by simple and multiple regression and regression ANOVA to determine possible correlations between them. RESULTS: VEGF and IL-6 concentrations were significantly lower within normal than within diseased gingiva. The number of blood vessel profiles and mean IL-6 concentrations were highest in diseased tissues adjacent to >6 mm sulci and were significantly correlated with sulcular depth (P <0.001). In contrast, VEGF concentrations were highest within diseased gingiva adjacent to 4 to 6 mm periodontal pockets (P <0.001) and were not correlated with sulcular depth. CONCLUSIONS: VEGF may be a factor in initiation and progression of gingivitis to periodontitis, possibly by promoting expansion of the vascular network coincident to progression of the inflammation. PMID- 10476892 TI - Use of lyodura for bone augmentation of osseous defects around dental implants. AB - BACKGROUND: Lyodura has been used in periodontology and maxillofacial surgery to overcome different clinical conditions. The use of lyodura to induce new bone formation in humans has not been widely reported. The purpose of this article is to describe the use of lyodura as a resorbable barrier to promote bone formation in osseous defects around dental implants. METHODS: The study group consisted of 22 healthy patients (12 women and 10 men), with a mean age of 32 (range 20 to 45). A total of 44 implants (9 titanium screw-type and 35 hydroxyapatite-coated cylinder implants) were placed. Of these, 27 implants were placed in the maxilla and 17 in the mandible. At 27 extraction sites bony vertical defects were measured on the buccal, lingual, mesial and distal sides of the implant. In the remaining 17 cases, where either immediate or staged implantation was performed, and a dehiscence was present, its vertical length was measured from the alveolar crest. All bony defects were grafted with autogenous bone harvested from the tuberosity. Each site was covered with lyodura. RESULTS: Healing was uneventful; no inflammation, infection, or soft tissue dehiscence was observed. At all extraction sites and in 76% of the dehiscence defects, hard tissue appearing clinically similar to bone completely filled the defect. Of those defects not completely filled, a mean vertical bone increase of 2.5 mm was recorded. A mean vertical bone increase of 2.6 mm was achieved at extraction sites and a mean increase of 2.79 mm was achieved at dehiscence sites. CONCLUSIONS: Lyodura can safely and effectively be used for guided bone regeneration at both extraction sites and for dehiscence defects. PMID- 10476893 TI - Microbiological response of localized sites with recurrent periodontitis in maintenance patients treated with tetracycline fibers. AB - BACKGROUND: Whether adjunctive tetracycline fibers can provide an additive effect to scaling and root planing in treating non-responsive sites in maintenance subjects is still controversial. Recolonization of the bacteria from untreated sites or from the extracrevicular region may explain the insignificant response to local therapy. The purpose of the present study was to evaluate the microbiological response of sites treated with tetracycline fibers combined with scaling and root planing. METHODS: The study was conducted in a split-mouth design. Thirty patients on maintenance therapy having at least 2 non-adjacent sites in separate quadrants with probing depths between 4 to 8 mm with bleeding on probing, or aspartate aminotransferase enzyme levels > 800 microIU in the gingival crevicular fluid, were treated with scaling and root planing plus tetracycline fibers or with scaling and root planing only. Subgingival plaque samples were collected at baseline, and 1, 3, and 6 months following treatment. A. actino-mycetemcomitans, C. rectus, B. forsythus, E. corrodens, F. nucleatum, P. gingivalis, and P. intermedia were detected by culture, immunofluorescence, or PCR technique. RESULTS: There was a reduction of total bacterial cell count, as well as of certain periodontal pathogens, following treatment. The prevalence of A. actinomycetemcomitans, B. forsythus, and P. gingivalis and the mean proportions of C. rectus, P. intermedia, F. nucleatum, and P. gingivalis decreased after therapy, but there was no statistically significant difference between the 2 treatment groups with respect to bacterial proportions or the number of positive sites. Besides, the pathogens could not be eliminated from the periodontal pocket, and recolonization of the pocket was noted at 3 months post treatment. CONCLUSIONS: Bacteria located within the cheek, tongue mucosa, saliva, or untreated sites may contribute to reinfection of the pockets and explain the insignificant response to local tetracycline therapy. PMID- 10476894 TI - The effect of postsurgical naproxen and a bioabsorbable membrane on osseous healing in intrabony defects. AB - BACKGROUND: Previous reports in the literature have shown that non-steroidal anti inflammatory drugs (NSAID) may affect osseous tissues by either stimulating or inhibiting bone formation. This effect can be drug specific and different NSAIDs may produce opposite results. There are also reports showing that NSAIDs inhibit bone loss due to inflammatory disease process. The purpose of this randomized, controlled, blinded, clinical investigation was to determine the effect of a one week course of postsurgical naproxen on the osseous healing in intrabony defects. METHODS: Twenty-four vertical osseous defects in 24 patients were treated with either a bioabsorbable membrane plus twice daily postsurgical naproxen 500 mg for one week (test or GPN group) or with a polylactide bioabsorbable membrane alone (control or GA group). Twelve patients were included in each group. Treatment was performed on either 2- or 3-wall or combination defects. All measurements were taken from a stent by a calibrated, blinded examiner and open measurements were repeated at the 9-month second stage surgery. Power analysis to determine superiority of naproxen treatment showed that a 12 per group sample size would yield 87% power to detect a 2.0 mm difference and 64% power to detect a 1.5 mm difference. RESULTS: Open defect measurements from baseline to 9 months showed a statistically significant (P < 0.05) mean defect fill of 1.96 +/- 1.27 mm and 2.04 +/- 1.71 for the GPN and GA groups, respectively. This corresponded to a mean defect fill of 42% and a mean defect resolution of approximately 75% for both groups. The differences between GPN and GA groups were not statistically significant (P > 0.05). Defect fill of > or = 50% was seen in 6 defects (50%) in the GPN group and in 5 defects (42%) in the GA group. CONCLUSIONS: The administration of postsurgical naproxen failed to produce osseous healing that was statistically superior to that obtained with polylactide bioabsorbable membranes alone. PMID- 10476895 TI - The effect of postsurgical antibiotics and a bioabsorbable membrane on regenerative healing in Class II furcation defects. AB - BACKGROUND: The purpose of this randomized, controlled, blinded, clinical investigation was to determine the effect of postsurgical antibiotics on osseous healing in Class II furcation defects. METHODS: Twenty-four Class II furcation defects in 24 patients were treated with either a polylactide bioabsorbable membrane, demineralized freeze-dried bone allograft (DFDBA) plus antibiotics (GBA or test group) or with a polylactide membrane and DFDBA alone (GB or control group). Twelve patients were included in each group. The antibiotic regimen consisted of ciprofloxacin 250 mg twice daily and metronidazole 250 mg tid for 1 week followed by a 7-week regimen of doxycycline hyclate 50 mg daily. Treatment was performed on either mandibular buccal or lingual, or maxillary buccal Class II furcation defects. Defects were randomly selected by a coin toss for treatment and all open and closed measurements were performed by a blinded examiner. Final open and closed measures from a stent were repeated at the 9-month second stage surgery. Power analysis to determine superiority of antibiotic treatment showed that a 12 per group sample size would yield 93% power to detect a 1.5 mm difference and 64% power to detect a 1 mm difference. RESULTS: Mean open horizontal probing depth reductions at 9 months were greater for the GBA group than for the GB group (2.92+/-1.78 versus 2.50+/-1.62 mm); however, these differences were not statistically significant. Seven of 12 furcations (58%) in the GBA group demonstrated >50% vertical defect fill at 9 months compared to 8 of 12 furcations (67%) in the GB group. There were no significant differences in mean open horizontal probing depth reduction between smokers and non-smokers in either the GBA or GB groups. Membrane exposure did not appear to affect regenerative healing in either the GBA or GB groups. CONCLUSIONS: The administration of postsurgical antibiotics did not produce statistically superior osseous healing of Class II furcation defects. This result may be attributable to membrane design which facilitates connective tissue ingrowth, thereby preventing bacterial downgrowth and contamination of the newly regenerated tissues. PMID- 10476896 TI - Resistance profile survey of 50 periodontal strains of Actinobacillus actinomyectomcomitans. AB - BACKGROUND: Antibiotic resistance has been increasingly described among bacterial species colonizing periodontal pockets, particularly in Prevotella and Porphyromonas spp. strains producing beta-lactamases, and frequently associated with resistance to tetracycline and erythromycin. These resistance genes may be carried on motile genetic elements, or transposons, capable of interspecies and intergeneric transmission among bacterial strains colonizing a same ecological niche. The aim of this prospective study was to determine the resistance profile of Actinobacillus actinomycetemcomitans and the prevalence of A. actinomycetemcomitans strains producing beta-lactamases in periodontal pockets. METHODS: Fifty strains of A. actinomycetemcomitans were isolated from 42 patients with adult periodontitis. No patient had periodontal or antibiotic therapy in the previous 6 months. Bacterial samples were collected from periodontal pockets > or =5 mm, appropriately diluted, inoculated onto selective medium (chocolate blood agar with bacitracin 75 microg/ml and vancomycin 5 microm/ml) and incubated for 5 days at 37 degrees C in air with 5% CO2. After conventional identification, susceptibility testing to 11 antibiotics was performed by the broth dilution method, in trypticase soy broth supplemented with yeast extract, hemin, and 0.1% NaHCO3 to maintain microaerophilic conditions in the microtitration plate wells by CO2 formation. RESULTS: No strain demonstrated resistance to amoxicillin, amoxicillin-clavulanic acid combination, pristinamycin, or ciprofloxacin at the breakpoint, but 40% of the strains were slightly resistant to penicillin G, and 4% were resistant to erythromycin, 90% to spiramycin, 18% to clarythromycin, 4% to tetracycline, 72% to metronidazole, and 12% to ornidazole. Amoxicillin, followed by tetracycline and erythromycin, was the most effective antibiotic on A. actinomycetemcomitans. The phenotypic research of a beta-lactamase was negative for all the strains tested. CONCLUSIONS: In this work, most A. actinomycetemcomitans strains were resistant to metronidazole, but the amoxicillin-metronidazole association may be of interest against subgingival anaerobic and capnophilic mixed flora. Pristinamycin and ciprofloxacin appeared as effective alternative monotherapies against A. actinomycetemcomitans. The threat of beta-lactam antibiotic resistance related to beta-lactamase production is currently not a problem with A. actinomycetemcomitans as it has been reported in oral anaerobes. PMID- 10476897 TI - Disposition of alendronate following local delivery in a rat jaw. AB - BACKGROUND: Recently, we have shown that local delivery of alendronate reduced significantly bone resorption activated by surgical separation of periosteum from bone. These results advocate the use of local application of alendronate in bone surgeries to prevent regional bone resorption at the surgery site. Here we investigated the efficacy of absorbtion of alendronate by the bone from a gelatin sponge soaked with radiolabeled alendronate applied topically at the surgical site. METHODS: Following elevation of the mucoperiosteal flap next to premolars and molars of the rat mandible, a gelatin sponge soaked with 10 microl of radiolabled alendronate (1 microCi/mg) was applied to exposed bone on one side. The local absorbtion of alendronate and its disposition in the contralateral side of the mandible as well as in the tibia bone were analyzed. RESULTS: The results show that 10% of total alendronate content of the gelatin sponge was absorbed in the bone locally (in the surgical site), while 0.2% was disposed in the tibia. Of interest is the fact that the surgical wound in the contralateral side increased the disposition of alendronate up to 2%. This finding is most likely the result of extravasation and diffusion of alendronate due to surgical wounding. CONCLUSION: This study strongly supports our notion that local delivery of alendronate and its affinity to bone may become a very important treatment modality to prevent resorption of bone during dental and orthopedic procedures. PMID- 10476898 TI - Long-term evaluation of 282 implants in maxillary and mandibular molar positions: a prospective study. AB - BACKGROUND: Placement of implants into molar positions presents diagnostic, surgical and prosthetic challenges. There are few reported studies for implants placed into molar positions. The purpose of this prospective longitudinal study is to report long-term clinical outcomes for 282 implants placed into molar positions. METHODS: Two-hundred-twelve patients received 282 implants. Implant size, location, jaw shape, and bone quality were recorded for all implants placed into molar positions. Seventy implants were inserted in maxillae and 212 in mandibles. Marginal bone level changes in maxillae and mandibles were measured from non-standardized periapical radiographs taken at abutment connection and an average follow-up of 3.9 years. Mesial-distal implant measurements were made from the top of the implant cylinder to the first point of bone to implant contact. In mandibles, 39 implants were used for single molar replacements, 67 implants were placed into excellent bone quality (type I) and 113 were in good bone quality (type 11); 145 implants were placed into bone with moderate bone resorption (type B); 166 implants were placed in first molar positions and 46 in second molar sites. RESULTS: At 6 years the cumulative success rate (CSR) for mandibular implants is 91.5%, and the success rate from the 2 to 3 year follow-up is 100%. Of the 70 implants placed in maxillae, 16 replaced single molars. Thirty-two implants were placed in jaw shape B with type 2-bone quality. For maxillary implants, the 6-year CSR was 82.9% and the success rate remained steady at 100% after the 2 to 3 year follow-up. For maxillary implants, at abutment connection the average marginal bone level was 1.67 mm, while at follow-up it was 1.98 mm. These differences were statistically significant (P = 0.04), but are not considered to be clinically significant. For mandibular implants, at abutment connection the mean marginal bone level as measured from radiographs was 2.11 mm, and at follow-up was 2.02 mm. This slight gain in bone level was not statistically significant and is not considered to be clinically significant. CONCLUSIONS: Results of this prospective longitudinal study of implants placed into molar positions indicates favorable clinical outcomes. These CSR rates (91.5% mandibles, 82.9% maxillae) are less than what has been reported for implants placed into mandibular and maxillary anterior segments. Differences in outcomes between anterior and posterior locations may be related to bone quality and quantity. PMID- 10476899 TI - Involvement of cyclooxygenase-2 in interleukin-1alpha-induced prostaglandin production by human periodontal ligament cells. AB - BACKGROUND: Human periodontal ligament (PDL) cells produce prostaglandin (PG) E2 in response to proinflammatory cytokines. However, the mechanism of PGE2 production is not well understood. The purpose of the present study was to investigate the involvement of cyclooxygenase (COX)-1 and COX-2 in PGE2 production by PDL cells stimulated with a proinflammatory cytokine, interleukin 1alpha (IL-1alpha), and to examine the regulation of PGE2 production by cell-cell interaction of human gingival keratinocytes and PDL cells. METHODS: The levels of PGE2 in the culture media of PDL cells stimulated with IL-1alpha or culture media of human gingival keratinocytes were determined by an enzyme-linked immunosorbent assay. Expression of COX-1 and -2 mRNA and protein was studied by Northern blot analysis and Western blot analysis, respectively. RESULTS: IL-1alpha-stimulated PDL cells produced PGE2 in a time-dependent manner. Indomethacin, a non-selective COX-1/COX-2 inhibitor, and NS-398, a selective COX-2 inhibitor, completely inhibited PGE2 production by the IL-1alpha-stimulated cells. COX-2 mRNA was detected after IL-1alpha stimulation, although it was not detected in unstimulated cells. There was no difference in expression of COX-1 mRNA between unstimulated cells and IL-la-stimulated cells. Expression of COX-2 protein in IL 1alpha-stimulated cells was increased, compared with that in unstimulated cells, whereas COX-1 protein expression was almost the same in both the cells. Treatment of IL-1alpha-stimulated PDL cells with dexamethasone, known to inhibit COX-2 expression, prevented PGE2 production and COX-2 mRNA expression. Addition of the culture media of human gingival keratinocytes to PDL cells increased PGE2 production. The PGE2 production was depressed by treatment of the cells with IL-1 receptor antagonist and anti- IL-1alpha antibody, not with anti-IL-1beta antibody. The PGE2 production was also inhibited by treatment with NS-398 and dexamethasone. CONCLUSIONS: We suggest that PDL cells stimulated with IL-1alpha produce PGE2 through de novo synthesis of COX-2 and that the cell interaction of gingival keratinocytes and PDL cells controls COX-2 expression and PGE2 production via IL-1alpha or 1alpha IL-la-like factor(s). Selective COX-2 inhibitors, which have the advantage of reduced gastric toxicity, may provide a useful approach to treatment of periodontal disease. PMID- 10476900 TI - Improvement of multiple facial gingival recession by non-surgical and supportive periodontal therapy: a case report. AB - We report a case of multiple facial gingival recession which was improved by non surgical and supportive periodontal therapy. The patient, a 28-year-old Japanese woman, presented for treatment of multiple facial gingival recession ranging from 1 to 4 mm on teeth 5 through 12 and 19 through 30. Periodontal plastic surgery to cover the exposed multiple root surfaces was suggested. However, because of emotional problems, the patient did not wish to undergo the procedures and instead accepted non-surgical periodontal therapy including oral hygiene instruction, scaling, and root planing. The exposed root sites were monitored at periodic maintenance visits, and gradual improvement through a coronal increase of the gingival margin was noted. The possible etiologic factors and healing process associated with this case are discussed. PMID- 10476901 TI - Gastric and gingival localization of mucosa-associated lymphoid tissue (MALT) lymphoma. An immunohistochemical, virological and clinical case report. AB - Non-Hodgkin's lymphomas (NHL) of the mucosa-associated lymphoid tissue (MALT) are characterized by their mucosal and glandular tissue localization. The case described here falls into the European-American classification of a low-grade B cell lymphoma of the MALT type, with a gingival lesion 2 years after a gastric lesion. The pathogenetic mechanisms of NHL in oral MALT and the diagnosis and treatment are discussed. PMID- 10476902 TI - Destructive membranous periodontal disease (Ligneous periodontitis). AB - Generalized membranous gingival enlargement due to accumulation of amyloid or fibrin-like material is a rare, destructive and poorly defined disease entity. Some patients also show extraoral manifestations. The lesion is an involvement of periodontal tissues caused by the same process as ligneous conjunctivitis. In this report, 3 new cases, two of whom are siblings, are presented. Defective fibrinolysis and abnormal wound healing seem to be the main pathogenetic mechanism of this unusual disease, which should be evaluated systemically considering other mucosal involvement. PMID- 10476903 TI - Rotated split palatal flap for soft tissue primary coverage over extraction sites with immediate implant placement. Description of the surgical procedure and clinical results. AB - Immediate implant placement after tooth extraction is becoming a common procedure in implant-supported oral rehabilitation. However, lack of primary full flap closure can jeopardize final results. A surgical approach that would enable predictable primary soft tissue closure over implants placed into fresh extraction sockets is described and evaluated. This technique is based on a rotated deep split thickness palatal flap (RSPF) containing periosteum and connective tissue, covering the implant and/or a barrier membrane. In 29 patients, 33 consecutive implants were placed immediately post extraction of 1 or 2 anterior or premolar maxillary teeth. Patients were divided in 2 groups: Group A (15 patients; n = 18 sites) where no barrier membrane was used and Group B (14 patients, n = 15 sites) where an occlusive resorbable collagen membrane was used. Distance between the alveolar crestal bone and the coronal aspect of the implant was measured at time of implant placement (Group A: mean 1.9 mm, SD 1.16; Group B: mean 4.6 mm, SD 1.18) and at second stage surgery (Group A: mean 0.3 mm, SD 0.46; Group B: mean 0. 7 mm, SD 0. 7). The difference between both records (crestal bone formation) was calculated (Group A: 1.7 mm, SD 1.03; Group B: 3.9 mm, SD 1.12) and found to be statistically significant (P <0.0001). Crestal bone formation, relative to the initial bone crest-implant distance at time of implant placement was approximately 85% in both groups. In 4 sites (2 in each group), where the cover screws were exposed before second stage surgery, complete crestal bone regeneration did not occur. Use of a barrier membrane may be obviated in appropriate cases while placing implants into fresh extraction sites. This procedure offers a predictable treatment approach in achieving complete soft tissue coverage, while allowing for healing of bony defects in immediate implantation procedures. PMID- 10476904 TI - Diabetes and periodontal diseases. AB - This position paper on diabetes mellitus was prepared by the Research, Science and Therapy Committee of The American Academy of Periodontology. It is intended to: 1) update members of the dental profession on the diagnosis and medical management of patients with diabetes mellitus; 2) summarize current knowledge on the relation between diabetes mellitus and periodontal diseases; 3) provide an overview of factors in diabetic patients relevant to understanding the pathogenesis of periodontal diseases in these subjects; 4) outline special considerations associated with treatment of periodontal diseases in diabetic patients; and 5) discuss possible approaches to the management of diabetic emergencies in the dental office. PMID- 10476905 TI - Variability in beta-adrenergic receptor population in cultured chicken muscle cells. PMID- 10476906 TI - Growth inhibition of human liver cancer cells by alpha-fetoprotein antisense strategy. PMID- 10476907 TI - Analysis of cytochrome P450 and phase II conjugating enzyme expression in adult male rat hepatocytes. AB - The induction of cytochrome P450 (CYP450) and Phase II conjugating enzymes by prototypical hepatic enzyme inducers was studied in adult male rat hepatocytes. Hepatocytes were suspended and cultured in diluted Matrigel in a basal serum-free Dulbecco's modified Eagle medium and exposed to the prototypical liver enzyme inducers, 3-methylcholanthrene, phenobarbital, hydrocortisone, and clofibrate for 48 h. Total RNA and microsomes were isolated and prepared, respectively, at 72 h. The expression of CYP1A1, CYP1A2, CYP2B1, CYP2C11, CYP2E1, CYP3A1, CYP3A2, CYP4A1, fatty acyl-CoA oxidase, uridine diphosphate-glucuronosyltransferase, glutathione-S-transferase, and sulfotransferase was determined at the mRNA level with reverse transcriptase polymerase chain reaction (RT-PCR). The expression of CYP1A1, CYP2B1, CYP2C11, CYP2E1, and CYP4A1 was also measured at the apoprotein level by Western immunoblotting. Using these culture and expression analysis techniques, we have found that the expression of these metabolic enzymes can be maintained in culture for up to 7 d at the mRNA and apoprotein levels. In addition, hepatocytes were found to respond to chemical enzyme inducers with marked increases in enzyme expression at either the mRNA or protein level and in a concentration-related fashion. Cells were responsive to enzyme induction as early as 24 h after initial plating. The results obtained from this investigation indicate that the presence of diluted Matrigel (at a concentration of 0.35 mg/ml), the use of low concentrations of insulin (1 microM), hydrocortisone (0.1 microM), and serum-free culture medium can maintain the differentiated phenotype and responsiveness of cultured hepatocytes to chemical-induced metabolic enzyme expression. Under the conditions used in this study, enzyme induction in adult male rat hepatocytes shows close agreement with enzyme induction observed in the livers of rats exposed to these or similar prototypical enzyme inducers. Rat hepatocytes cultured in the presence of diluted Matrigel coupled with enzyme mRNA expression analysis with RT-PCR are proven to be a valuable and important in vitro toxicological approach to assess the chemical-induced changes in expression of liver CYP450 and Phase II conjugating enzymes. PMID- 10476908 TI - Effects of hepatocyte growth factor (HGF) and activin A on the morphogenesis of rat submandibular gland-derived epithelial cells in serum-free collagen gel culture. AB - To study the mechanisms of morphogenesis in salivary gland regeneration, we have established the RSMG-1 cell line derived from submandibular gland (SMG) of 10-wk old Wistar female rats in serum-free culture. Our finding that RSMG-1 cells originated from duct cells was based on morphology and immunohistochemical results. In three-dimensional serum-free collagen gel culture, HGF induced branching morphogenesis of RSMG-1 cells. Histological examination revealed that HGF-induced branching structure exhibited well-formed lumina. This morphology closely resembles that found in vivo. The cells also expressed activin A. Exogenously added activin A at a high concentration reduced HGF-induced branching morphogenesis. These findings suggest that the morphogenesis of the salivary gland is modulated by HGF and activin A. Our results show that the RSMG-1 cell line may be useful in studies of salivary gland regeneration. PMID- 10476909 TI - Growth and phenotypic characterization of porcine coronary artery smooth muscle cells. AB - Vascular smooth muscle cell (VSMC) proliferation significantly contributes to atherosclerotic plaque formation and limits the success rate of percutaneous transluminal coronary angioplasty. We derived a population of porcine coronary artery SMCs to characterize VSMC proliferation and phenotype in preparation to study the molecular actions of VSMC mitogens and antiproliferative agents. Growth assays were designed to minimize the estrogen content in the culture medium, since this steroid hormone significantly influences VSMC growth and the expression of VSMC mitogens and their receptors. Culture conditions were identified such that this criterion was achieved while maintaining a significant VSMC growth rate. Cells cultured in serum-free medium, regardless of growth factor supplements, did not remain adherent to a plastic culture substrate, nor did they proliferate. Dextran-coated charcoal (DCC)-treated sera, including fetal bovine, calf, and porcine, supported VSMC adhesion, but not growth. Whole fetal bovine serum (FBS) produced the best proliferative response. A type-I collagen coated culture surface significantly enhanced VSMC growth, but only in culture medium containing non-DCC-treated FBS. Flow cytometry analyses confirmed the mitogenic effects of this substrate. The VSMCs exhibited a morphological change on type-I collagen, but this was not accompanied by a change in VSMC phenotype. Our data indicate that culture of these porcine coronary artery SMCs in 2.5% FBS plus 10 ng platelet-derived growth factor-BB per ml in phenol red-free medium on type-I collagen may be the optimal conditions for studying the molecular aspects of VSMC mitogens and antiproliferative agents. PMID- 10476910 TI - Alteration of cytoskeletal structure, integrin distribution, and migratory activity by phagocytic challenge in cells from an ocular tissue--the trabecular meshwork. AB - The trabecular meshwork is a specialized tissue in the anterior chamber of the eye that regulates the aqueous humor outflow and controls the intraocular pressure. Cells in the trabecular meshwork are believed to be essential for maintenance of the outflow system, and their malfunctioning may lead to elevation of intraocular pressure and development of glaucoma. These cells are avid phagocytes. Using an in vitro tissue culture system, we have previously shown that bovine trabecular meshwork cells exhibited a short-term loss of cell-matrix adhesiveness after exposure to latex microspheres. The current study showed that 4 h after phagocytosis, the cytoskeletal structure in trabecular meshwork cells was disrupted, the formation of focal contact formation was limited, and the cellular migratory activity was increased. These in vitro responses paralleled those that occur in vivo. By 24 h, all the changes demonstrated returned to normal. Our data suggest that the short-term loss in cell-matrix cohesiveness observed after phagocytic challenge may be related to the reorganization of cytoskeletal structures and the decline of focal contact formation. The altered cell migration may also be interlinked. PMID- 10476911 TI - N5 endometrial stromal cell line: a model system to study decidual prolactin gene expression. AB - Prolactin gene expression in extrapituitary tissues, such as decidua and lymphocytes, is regulated by a distinct promoter approximately 6 kb upstream of the pituitary prolactin gene transcription start site. Here we describe studies in a human endometrial stromal cell line, N5, that was immortalized by transfection with an SV40 mutant and which expresses the prolactin gene driven by the extrapituitary promoter. The N5 cells have phenotypic features of primary cultures of decidualized human endometrial stromal cells and secrete low levels of prolactin and insulin-like growth factor binding protein-1 (IGFBP-1), both of which are markers of decidualized endometrial stromal cells. As in primary cultures of endometrial stromal cells, treatment of N5 cells with progesterone and estradiol alone or in combination with prostaglandin E2 stimulated the synthesis and release of prolactin. Transient transfection of the N5 cells with an expression vector containing - 2927/ + 66 bp of the decidual prolactin promoter coupled to a luciferase reporter gene resulted in a 20 to 25-fold increase in luciferase activity, a magnitude similar to that which occurs in primary cultures of endometrial stromal cells decidualized in vitro by treatment with progesterone and estradiol. Luciferase expression levels were similar in untreated N5 cells and N5 cells treated with progesterone and estradiol. Taken together, these results indicate that the N5 human endometrial stromal cell line has phenotypic characteristics of normal decidualized stromal cells and is a useful model to study regulation of decidual prolactin gene expression. PMID- 10476912 TI - Establishment and cellular characteristics of a hepatocyte cell line (OUMS-31) derived from an acatalasemic mouse. AB - Liver cell lines with very low catalase activity were established from an acatalasemic mouse. Hepatocytes isolated by a collagenase-liver-perfusion technique were cultured in Williams' E medium supplemented with 10% fetal bovine serum. The acatalasemic liver cell line showed approximately 20% of the catalase activity of a normal mouse liver cell line, whereas its glutathione peroxidase activity was approximately equal to that of the normal liver cell line. DNA sequence analysis of this cell line showed the same mutation in the catalase gene as is seen in the acatalasemic mouse. Our observation of intracellular content of hydrogen peroxide (H2O2) radical and increased susceptibility of the cells to H2O2 were compatible with the existence of low catalase activity in the acatalasemic mouse. This hepatocyte cell line should be useful for studying effects of oxidative radical stress at the cellular level. PMID- 10476913 TI - Initial aspects of mineralization at the dentino-enamel junction in embryonic mouse incisor in vivo and in vitro: a tem comparative study. AB - The frontier between the enamel organ and the dental papilla, the future dentino enamel junction, undergoes coordinated modifications. The mineralization of the extracellular matrix starts within the predentine, which is a prerequisite for the formation of the first enamel crystallites in vivo. We investigated the dentino-enamel junction using the embryonic mouse incisor as a model. Our data showed that the notion of the dentino-enamel junction should not be restricted to the thin interface classically described. A temporo-spatial survey from the epithelio-mesenchymal junction to the dentino-enamel junction delineated a clear sequence of events characterized by the early deposition of electron-dense granules, followed by the appearance of patches of stippled material at the dentino-enamel junction. The first tiny enamel crystallites appeared in the vicinity of this material which presented a well-ordered alignment. The comparison of data obtained in vivo on 17-, 18-, 19-d-old embryonic incisors with those obtained in vitro using 15-d-old embryonic incisors cultured for 7 d emphasizes the relevance of this sequence. Helicoidal growing crystals were observed in cultured tooth germs but never in vivo. PMID- 10476914 TI - Structure-activity relationship of polyphenols on inhibition of chemical mediator release from rat peritoneal exudate cells. AB - The effect of phenolic compounds in foodstuffs on histamine and leukotriene B4 (LTB4) release from rat peritoneal exudate cells and their antioxidative activity were examined to assess their antiallergenic activities. Among them, triphenols such as pyrogallol and gallic acid inhibited histamine release from the cells, but diphenols did not. On the other hand, o- and p-diphenols such as catechol and hydroquinone with strong antioxidative activity inhibited LTB4 release as strongly as pyrogallol, but an m-derivative resorcinol with weak antioxidative activity did not. Though carboxylated compounds and their noncarboxylated counterparts were antioxidative, the former exerted a much weaker inhibitory effect on the LTB4 release than the latter. In flavonols, only myricetin with a triphenolic B ring strongly inhibited histamine release, but all flavonols strongly suppressed LTB4 release irrespective of the number of OH groups in the B ring. Among flavonoids with an o-diphenolic B ring, flavonol and flavone with a C4-carbonyl group strongly inhibited LTB4 release, whereas the activity of anthocyan without C4-carbonyl was much weaker than the above compounds. These results suggest that triphenolic structure is essential for the inhibition of histamine release. On the other hand, antioxidative activity and membrane permeability of phenolic compounds seemed to be essential for the inhibition of LTB4 release. In addition, the C4-carbonyl group seemed to be important for strongly inhibiting LTB4 release. PMID- 10476916 TI - Stable transformation of lepidopteran cultured cells with novel cloning method. PMID- 10476915 TI - New adipogenic cell lines derived from C3H10T1/2. PMID- 10476917 TI - Toxicity of ifosfamide and its metabolite chloroacetaldehyde in cultured renal tubule cells. AB - Renal injury is a common side effect of the chemotherapeutic agent ifosfamide. Current evidence suggests that the ifosfamide metabolite chloroacetaldehyde may contribute to this nephrotoxicity. The present study examined the effects of ifosfamide and chloroacetaldehyde on rabbit proximal renal tubule cells in primary culture. The ability of the uroprotectant medication sodium 2 mercaptoethanesulfonate (mesna) to prevent chloroacetaldehyde-induced renal cell injury was also assessed. Chloroacetaldehyde (12.5-150 microM) produced dose dependent declines in neutral red dye uptake, ATP levels, glutathione content, and cell growth. Coadministration of mesna prevented chloroacetaldehyde toxicity while pretreatment of cells with the glutathione-depleting agent buthionine sulfoximine enhanced the toxicity of chloroacetaldehyde. Ifosfamide (1000-10,000 microM) toxicity was detected only at concentrations of 4000 microM or greater. Analysis of media collected from ifosfamide-treated cell cultures revealed the presence of several ifosfamide metabolites, demonstrating that renal proximal tubule cells are capable of biotransforming this chemotherapeutic agent. This primary renal cell culture system should prove useful in studying the cause and prevention of ifosfamide nephrotoxicity. PMID- 10476918 TI - Characterization of a new tissue-engineered human skin equivalent with hair. AB - We designed a new tissue-engineered skin equivalent in which complete pilosebaceous units were integrated. This model was produced exclusively from human fibroblasts and keratinocytes and did not contain any synthetic material. Fibroblasts were cultured for 35 d with ascorbic acid and formed a thick fibrous sheet in the culture dish. The dermal equivalent was composed of stacked fibroblast sheets and exhibited some ultrastructural organization found in normal connective tissues. Keratinocytes seeded on this tissue formed a stratified and cornified epidermis and expressed typical markers of differentiation (keratin 10, filaggrin, and transglutaminase). After 4 wk of culture, a continuous and ultrastructurally organized basement membrane was observed and associated with the expression of laminin and collagen IV and VII. Complete pilosebaceous units were obtained by thermolysin digestion and inserted in this skin equivalent in order to assess the role of the transfollicular route in percutaneous absorption. The presence of hair follicles abolished the lag-time observed during hydrocortisone diffusion and increased significantly its rate of penetration in comparison to the control (skin equivalent with sham hair insertion). Therefore, this new hairy human skin equivalent model allowed an experimental design in which the only variable was the presence of pilosebaceous units and provided new data confirming the importance of hair follicles in percutaneous absorption. PMID- 10476919 TI - Spontaneous activation of endothelial cells: a central role for endogenous IL 1alpha. AB - Endothelial cells assume divergent physiologic postures, from a 'quiescent' to an 'activated' state, and this ability is thought to critically regulate homeostasis of the vascular milieu. While establishing endothelial cell lines, we observed that most of the endothelial cells exhibited 'spontaneous' activation with morphological changes once the cells surpassed confluence. In the present study, we investigated whether accumulation and subsequent availability of certain factors in the cells might induce morphologic changes indicating activated phenotype in confluent endothelial cells. Cell lysate from nonactivated confluent endothelial cells activated autologous target cells, whereas culture supernatants did not. Stimulatory activity of the cell lysate was dependent on the concentration of cell lysate, i.e., nonactivated endothelial cell lysate at 25% concentration induced a substantial level of morphologic change. The 'spontaneous' as well as the cell lysate-induced change in morphology of endothelial cells was inhibited by neutralization of interleukin (IL)-1alpha with anti-IL-1alpha antibody. Correspondingly, cell lysate from confluent non-IL-1 expressing endothelial cells did not alter the morphology of autologous confluent cells even at a higher concentration. Cells that spontaneously changed their morphology correspondingly upregulated IL-1alpha and IL-8 transcripts. The results indicate that the accumulation and availability of endogenous IL-1alpha is responsible for the 'spontaneous' activation of endothelial cells in culture. Ultimately, through this property, endothelial cells may facilitate normal in vivo host responses, including the regulation of leukocyte trafficking. PMID- 10476920 TI - The establishment of new cell lines from Pseudaletia unipuncta with differential responses to baculovirus infection. AB - Six insect cell lines from Pseudaletia unipuncta embryos were established and characterized, and their susceptibility to Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) infection was investigated. These embryonic P. unipuncta cell lines had characteristics distinct from each other in morphology and growth, and showed differential responses to AcMNPV infection. Among the six cell lines, two were highly susceptible to virus infection. One of these two cell lines, BTI-Pu-A7S, produced over 100 AcMNPV occlusion bodies per cell, on average. Three cell lines showed an apoptotic response following AcMNPV infection. One cell line did not support complete virus replication through the late phase of virus growth and did not exhibit apoptosis. The P. unipuncta cell lines could be distinguished from SF21 and BTI-Tn-5B1-4 cells by their isozyme markers. PMID- 10476921 TI - Characterization of a cell line established from diethylstilbestrol-induced renal tumors in Syrian hamsters. AB - This article describes HKT-1097, a new cell line established from renal tumors induced by the protracted administration of diethylstilbestrol (DES) to male Syrian golden hamsters. Cell culture was initiated from tumor samples obtained from two 14-mo.-old animals which had undergone exposure to DES for a period of 11 mo. The HKT-1097 cell line was characterized between Passages 16 and 22 with respect to cell morphology, growth properties, karyology, and the presence of estrogen receptors. Moreover, immunostaining with a panel of antisera was performed to identify the cytological profile of the cell line and establish a parallel with tumor tissue in vivo. HKT-1097 cells are fibroblastoid; their most distinctive feature is that they exhibit strikingly long processes. The HKT-1097 cell line grows as a monolayer with a tendency toward a less stringent density dependent inhibition of growth. The modal chromosome number is 44, but more than 50% of the cells are aneuploid, suggesting a substantial degree of karyotype instability. HKT-1097 cells express estrogen receptors. They contain immunoreactive vimentin and desmin, but appear negative upon cytokeratin immunostaining. In addition, these cells express glial fibrillary acidic protein and other markers of the neuroectodermal lineage, but lack neurofilament protein. Insofar as the same lineage markers have been demonstrated in DES-induced Syrian hamster kidney tumors (SHKT), we conclude that HKT-1097 cells retain some of the original tumor cell phenotype. The current observations suggest that estrogen induced SHKT derive from the renal interstitium and point to an involvement of neuroectodermal cells in the development of these neoplasms. PMID- 10476922 TI - Human endothelial cells cultured on microporous filters used for leukocyte transmigration studies form monolayers on both sides of the filter. AB - A growing number of studies on the mechanism of leukocyte transendothelial migration use endothelial cells grown on microporous filters as an in vitro model of endothelium. Ultrastructural examination of such a model system previously demonstrated that human pulmonary artery endothelial cells (HPAEC) formed confluent monolayers on both sides of the 3-microm-pore filter (Mackarel et al., 1999). To determine whether this was a characteristic specific to pulmonary artery endothelial cells, the growth characteristics of a human pulmonary microvascular endothelial cell type (HMVEC-L) and the widely used human umbilical vein endothelial cells (HUVEC) on 3-microm microporous filters were examined by transmission electron microscopy (TEM). Similar to HPAEC, HMVEC-L and HUVEC were also found to grow on both sides of the filter. All three endothelial cell types were capable of migrating through the 3 microm pores of the filter to form a monolayer on the filter underside. The endothelial cells on the underside were orientated in an inverted position with the luminal surface facing away from the filter. Such 'bilayer' formation was observed at a range of seeding densities and in different culture media. Despite the presence of a bilayer of endothelial cells, TEM demonstrated that neutrophils migrated successfully across the cell filter-cell system. Previous transmigration reports in which an in vitro model similar to ours was used have often assumed only one layer of endothelial cells. The observations reported here indicate that while endothelial cells on microporous filters are useful models for examining leukocyte-endothelial interactions, they are not appropriate for studies examining endothelial cell 'sidedness.' PMID- 10476923 TI - Bioassay development: the implications of cardiac myocyte motility in vitro. AB - Cardiac myocytes cultured over microfabricated extracellular recording devices can be used to assay bioactive compounds. However, electrophysiological signals recorded from these devices vary in amplitude with time. Theoretically, changes in signal amplitude arise from myocytes being moved over recording sites by cocultured fibroblasts. To test this, neonatal rat cardiac myocytes were cultured at high densities and low densities on fibronectin-coated glass. After 36.5 h, myocytes were identified by their rhythmic contractions and then time-lapse recorded for 3.5 h. Length, width, and angle of orientation was then determined every 30 min for five cells in low density and five cells in high-density culture. Low-density cells had mean lengths of 65.3 microm and widths of 35.1 microm, whereas cells in high-density culture had greater mean lengths of 74.2 microm and lower mean widths of 24.3 microm. Length, width, and angle of orientation of cells in low- and high-density culture changed by 4.1%, 11.8%, and 2.7 degrees, and 6.4%, 10%, and 4.6 degrees, respectively, every half hour. We found no evidence of myocyte-fibroblast interactions influencing cell position or shape in low density, but in high density, we found evidence that fibroblast myocyte interactions could transiently influence cell shape. We conclude that fibroblast-independent changes in cell shape are largely responsible for the changes in signal amplitude recorded from cardiac myocytes cultured on microfabricated extracellular recording devices. However, there is some evidence that myocyte-fibroblast interactions may augment this process in high-density culture. The implications of these findings for bioassay development are discussed. PMID- 10476925 TI - Immunomodulation and therapeutic effects of the oral use of interferon-alpha: mechanism of action. AB - It is now well accepted that type 1 interferons (IFNs), IFN-alpha and IFN-beta, in addition to being molecules with powerful antiviral activity, play a critical role in modulating immune responses to foreign and self-antigens. This review of the literature documents the immunomodulatory effects of IFN-alpha and discusses its position and importance in the cytokine cascade. In addition, this review attempts to organize the literature describing local and systemic immunomodulatory effects of orally administered low doses of IFN-alpha, and provide a physiological explanation for the mechanism of action. Evidence suggests that, early in the process of antigen presentation to T helper (Th) cells, IFN-alpha derived principally from the antigen-presenting cells (APC) provides an important signal for Th precursor differentiation in favor of a Th1 immune response. IFN-alpha, perhaps via upregulation of the high-alphaffinity interleukin-12beta1/beta2 (IL-12beta1/beta2) receptor, renders Th1 cells responsive to IL-12 resulting in production of high levels of IFN-gamma crucial to the development of Th1 immune responses. In addition to being instrumental in the development of Th1 immune responses, IFN-alpha appears to be the major cytokine responsible for the amplification of the CD8+ T cell response and resistance to viral infections. Orally administered IFN-alpha induces similar Th1 cytokine responses in buccal mucosal lymph nodes (LN), including upregulation of IFN-gamma expression and downregulation of IL-4. Moreover, reports of systemic immune effects such as decreased autoimmune responses, increased antiviral and antibacterial responses, and generalized immune function changes after oral IFN alpha administration are consistent with the known immunomodulatory role of IFN alpha in a physiological setting. Responses to orally administered low doses of IFN-alpha also adhere to the principle of low-dose priming and high-dose anergy that dictates the cellular and cytokine responses to exogenously added cytokines both in vivo and in vitro. These observations collectively suggest that IFN-alpha administered to mucosal-associated immune tissue replicates the known physiological role of IFN-alpha, including regulation of CD4+ Th1 immunomodulatory cells and activation of CD8+ effector cells, which are both crucial to development of protective immune responses. What remains to be determined is how local mucosal immune responses to IFN-alpha given orally are translated into systemic immune responses and resistance to disease. This important question, the answer to which will have profound implications for new immunotherapies for immune-based diseases, is the focus of current research. PMID- 10476924 TI - The presence of transcription factors in chicken albumin, yolk and blastoderm. AB - Embryonic development is determined by preset intrinsic programs and extrinsic signals. To explore the possibility that transcription factors are present at the onset of development, preparations of yolk, albumin, and blastoderm from unfertilized and fertilized white Leghorn chicken eggs were screened by a panel of 16 transcription factor antibodies with Western blot techniques. Yolk was positive for 13 transcription factors, whereas blastoderm was positive for 10, and albumin was positive for 5. In yolk, several transcription factors, GATA-2, E2F-1, MyoD, and TFIID, were developmentally regulated. These results indicate that intracellular yolk and extracellular albumin contain transcription factors which presumably influence early chick embryonic development from prefertilization to the late blastoderm stage. Thus, the utility of preset maternal transcription factors within yolk and albumin complement maternally derived mRNA to determine the early development of the zygote. PMID- 10476926 TI - Systemic effects of orally administered interferons and interleukin-2. AB - Orally administered interferons (IFN-alpha, IFN-beta, and IFN-gamma) have been shown to exert a number of systemic effects. Orally administered IFNs exert dose dependent suppressive effects on the peripheral white blood cell (WBC) count. The suppression of the peripheral WBC count is mediated by a suppression of the function of the bone marrow, as measured in an in vitro bone marrow colony forming assay. The peripheral WBC and bone marrow suppressive effects of orally administered IFNs are at least as potent as those occurring with parenterally administered IFNs. However, the mechanism by which orally administered IFNs exert these peripheral WBC suppressive and bone marrow suppressive effects differs significantly from that of parenterally administered IFNs: orally administered IFN is not detectable in the serum, the effect of orally administered IFN is not blocked by circulating antibody, the effect of orally administered IFN can be adoptively transferred by injection with peripheral white blood cells from donor mice, and the effect of orally administered IFN develops more slowly than that of parenterally administered interferon. Orally administered IFN-alpha employed alone and in synergistic combination with intraperitoneally administered IFN gamma can exert an antitumor effect. Finally, orally administered interleukin-2 can exert a suppressive effect on both the peripheral white blood cell count and on the bone marrow. These observations suggest that the oral route may be an effective and novel mechanism for the efficacious administration of IFNs and other lymphokines/cytokines. PMID- 10476927 TI - Autoimmunity is a type I interferon-deficiency syndrome corrected by ingested type I IFN via the GALT system. AB - Type I interferons (IFN-alpha/beta), products of the innate immune system, can modulate immune function whereas proinflammatory IFN-gamma (type II IFN), a product of the acquired immune system upregulates inflammation and enhances cell mediated immunity. We have proposed a unifying hypothesis of the origin of autoimmunity as a type I IFN immunodeficiency syndrome involving inadequate regulation of the acquired immune system product IFN-gamma by the IFN-alpha/beta innate immune system. The common theme of ingested type I IFNs in autoimmunity is inhibition of proinflammatory type II IFN systemically or at the target organ. In multiple sclerosis (MS) and insulin-dependent diabetes mellitus (IDDM) at the target organ, and in rheumatoid arthritis (RA) as a regulator of other proinflammatory cytokines, IFN-gamma is the nexus of inflammation in autoimmunity. Ingested type I IFNs counteract type II IFN, overcome the relative lack of type I IFN activity, and ameliorate autoimmunity. The administration of type I IFNs (IFN-alpha/beta) via the gut offers an exciting alternative to systemic application for overcoming the type I IFN immunodeficiency in autoimmunity. Successful use of ingested type I IFN in three separate prototypical autoimmune diseases suggests a broad antiinflammatory therapeutic profile for this technology. PMID- 10476929 TI - The oropharyngeal delivery of interferons: where are we and where do we need to go? AB - The controversial concept of oral delivery of interferon (IFN) is reviewed in relation to possible mechanisms of action. Although there is a rational basis and some positive clinical results for believing that it may work, problems remain to understand which is the most effective IFN preparation and why. Owing to a minimal cost and lack of toxicity, the oral as well as other unconventional routes of IFN delivery deserve to be evaluated because they may become useful. PMID- 10476928 TI - Oral use of interferon. AB - Interferon-alpha (IFN-alpha) given orally has biological activity in humans and other animals. The dose providing the most benefit delivers IFN-alpha to the oral mucosa in a concentration (10(2)-10(3) IU), similar to that naturally produced in the nasal secretions during respiratory infections. In contrast, conventional IFN therapy employs parenteral doses of > 10(6) IU and, for this reason, orally administered IFN therapies have been called low-dose treatments. Efficacy in both animal disease models and human studies has been reported, and the mechanisms whereby oral administration has a systemic effect are under active study in a number of laboratories. PMID- 10476930 TI - Low-dose oral use of interferon inhibits virally induced myocarditis. AB - Cytomegalovirus (CMV) infection has been associated with the development of myocarditis in humans. Our established mouse model for CMV myocarditis allows detailed investigation of the immunopathogenic mechanisms and therapies for cardiovascular disease. The type I interferons (IFN-alpha/beta) are part of the innate immune response to CMV infections. Previously, we have reported that daily treatment with low doses of murine IFN-alpha/beta administered by the oral mucosal route significantly reduces early virus replication of murine CMV in the spleen and liver of infected mice. The oral-mucosal route provides an alternate delivery system to the current modes of IFN administration and is associated with fewer side effects. Since prophylactic treatment with type 1 IFNs may result in both antiviral and immunomodulatory effects that may lessen the development of disease, we wished to study the effect of IFN-alpha/beta on the development of myocarditis. Low-dose oral use of type I IFN (10 IU/day for 7 days prior to virus infection) did not abrogate myocarditis but suppressed the inflammatory response in both the acute and chronic phase of the disease. Furthermore, low-dose oral use of IFN was as effective at inhibiting myocarditis as a single injection of a high dose of IFN (20,000 IU) on the day of virus infection. These findings indicate the need for evaluation of low-dose use of oral IFN in the development of improved clinical therapies for the treatment of cardiovascular disease. PMID- 10476931 TI - Efficacy of low-dose oral use of type I interferon in cytomegalovirus infections in vivo. AB - Oral administration of type I interferons (IFNs; murine IFN-alpha and IFN-beta) reduces early replication of murine cytomegalovirus (MCMV) in both the spleen and liver of MCMV-infected BALB/c mice. Examination of a range of doses of IFN (1 to 1000 IU) showed that 10 IU administered daily for 1 week prior to virus infection was optimal for inhibition of MCMV replication. Furthermore, low-dose orally administered IFN (10 IU/day) was effective in mice challenged with lethal and sublethal virus inocula. The antiviral efficacy of low-dose orally administered IFN was not restricted by either the route of virus inoculation or the mouse genotype. Analysis by immunohistochemistry of IFN-alpha receptor-bearing cells of the gastrointestinal tract revealed predominant staining of perivascular smooth muscle and the lamina propria of the anterior tongue, small intestine and rectum. These tissues, dense in IFN-alpha receptor-bearing cells, are likely to be the sites of interaction of the orally administered IFNs with the mucosal immune system. In conclusion, we propose that low-dose oral use of type I IFN therapy may have broad applications in the treatment of CMV infections. PMID- 10476932 TI - Oral use of interferon-alpha delays the onset of insulin-dependent diabetes mellitus in nonobese diabetes mice. AB - Insulin-dependent diabetes mellitus (IDDM) in the nonobese diabetes (NOD) mouse model is thought to be an autoimmune CD4 Th1-like cell-mediated disease. We tested the efficacy of oral use of interferon-alpha (IFN-alpha) therapy on IDDM in NOD mice. Using urine and blood sugar levels as indicators of IDDM, oral administration of murine IFN-alpha (100 IU/body) to NOD mice significantly delayed the onset of symptomatic diabetes. However, oral use of IFN-alpha did not prevent diabetic NOD mice from losing weight once NOD mice were symptomatic, suggesting that orally administered IFN-alpha is a prophylactic rather than therapeutic approach to the management of IDDM. PMID- 10476933 TI - Interferon action on avian viruses. I. Oral administration of chicken interferon alpha ameliorates Newcastle disease. PMID- 10476934 TI - Suppression of late asthmatic response by low-dose oral administration of interferon-beta in the guinea pig model of asthma. AB - We investigated the anti-asthmatic effects of low-dose oral and subcutaneous administration of interferon-beta (IFN-beta) on an ovalbumin (OVA)-sensitized and challenged guinea pig model of asthma. Subcutaneous administration of IFN-beta suppressed the eosinophil infiltration by 14.2% of the control and the respiratory resistance (Rrs) by 58.2% at 2.0 MIU/kg. Oral administration of IFN beta inhibited the late asthmatic response (LAR) by suppressing the increase of Rrs by 29% of the control at 1 IU/ml and the eosinophil infiltration into the trachea and lung by 34.6% at the optimum dosage of 10 IU/ml. Both subcutaneous and oral administration could not inhibit the early asthmatic response (EAR). Additionally we found 2',5'-oligoadenylate synthetase (2',5'-OAS) induction by low-dose oral administration (LDOA) of IFN-beta to the same extent as by subcutaneous administration in whole blood in vivo. These data suggest that LDOA of IFN-beta would have some clinical benefit for patients with asthma. PMID- 10476936 TI - Low-dose oral administration of interferon-alpha for the treatment of immune mediated keratoconjunctivitis sicca in dogs. AB - This preliminary study was designed to evaluate the effectiveness and dosage of oral use of interferon-alpha (IFN-alpha) in the treatment of naturally occurring, immune-mediated, canine keratoconjunctivitis sicca (KCS). Dogs with chronic immune-mediated KCS were selected from the two clinic populations. All medication, except topical artificial tears, was discontinued at least 2 weeks prior to beginning the clinical trial. IFN-alpha was administered orally once daily to the dogs by their owners as the sole therapy for the KCS. Examinations of the dogs were performed every 2 weeks for the duration of the trial (12 weeks). Each dog was given either two or three separate, escalating doses (20, 40, 80 IU of the IFN-alpha. A favorable response was observed in 55% (11/20) of all dogs treated. Clinical findings of those dogs that responded included increased wetting of the eyes, decreased mucus discharge, and fewer signs of discomfort. There was a nearly significant difference (p = 0.08) in pretreatment mean Schirmer's tear test (STT) between the dogs that responded (6.4 +/- SEM 0.62 mm/min) and those that did not respond (4.7 +/- SEM 0.69 mm/min) to the orally administered IFN-alpha. Seven of 11 dogs with favorable outcomes had an increased STT of at least 5 mm/min after treatment with oral IFN-alpha and the group had a post-treatment STT (10.5 +/- SEM 1.4 mm/min) significantly greater than baseline (p = 0.0004). The post-treatment STT of the dogs that did respond was significantly greater (p < 0.01) than the post-treatment mean STT of dogs that did not respond. All dogs that responded did so with the 20 or 40 IU dose of IFN alpha. No side effects were noted and all dogs tolerated the treatment well. PMID- 10476935 TI - Effects of oral administration of interferon-alpha on antibody production in mice with induced tolerance. AB - In vivo systemic effects and the immunomodulating potential of the oral administration of murine interferon-alpha (IFN-alpha) were investigated through mRNA expression of both IFN-alpha-inducible factors, interferon regulatory factor 1 (IRF-1) and 2,5-adenylate synthetase [2-5(A) synthetase] and 2-5(A) synthetase enzymatic activity in spleen and antibody production. The daily administration of IFN-alpha (0.1, 1, 10, and 100 IU/body) for 1 week augmented IRF-1 and 2-5(A) synthetase mRNA expression levels, as well as 2-5(A) synthetase enzymatic activity in spleen cells but not in cervical lymph nodes. The in vivo immunomodulating potential of the oral administration of IFN-alpha was also evaluated through antibody production in mice with induced tolerance. Ovalbumin (OVA) was administered intraperitoneally (i.p.) to induce systemic antibody production on day 0 when OVA feeding was initiated. The OVA was fed every 2-3 days for a total of 14 doses to suppress serum antibody levels. Oral administration of murine IFN-alpha was initiated on day 0 and was continued for 5 consecutive days weekly for 5 weeks (24 doses). On every sampling date (days 10, 17, 24, and 32), specific antibody levels in the IFN-alpha-administered groups were significantly higher than those in the control (nonadministered) group. This was especially noted in early phases (days 10 and 17) of antibody production when the levels of antibody in serum from the IFN-alpha-administration groups were equivalent to those of the nontolerance group. Altogether, it is suggested that oral use of IFN-alpha can elicit immunomodulating actions (e.g., antibody levels) by affecting the systemic immune system(s). PMID- 10476937 TI - Natural human interferon-alpha administered orally as a treatment of bovine respiratory disease complex. AB - Natural human interferon-alpha (nHuIFN-alpha) from three sources was given orally to 368 calves experiencing a natural outbreak of bovine respiratory disease complex (BRDC). In one study, 200 calves were given one treatment daily for 3 days of placebo or 20, 200, or 2,000 IU of nHuIFN-alpha before shipment. Calves treated with 20 or 200 IU had a significant (p < 0.05) weight gain benefit for the first 21 days in the feedlot, if they had rectal temperatures <40 degrees C when treated with nHuIFN-alpha. In a second trial, 168 mixed-breed calves (five groups randomized to 31-36 calves/group) were treated with one dose of placebo or 200 or 400 IU of nHuIFN-alpha after shipment to the feedlot. Using this regimen, a dose of 200 IU per calf significantly (p < 0.08) decreased the number of sick calves per group and delayed development of BRDC. Results of these studies demonstrate that oral administration of nHuIFN-alpha, which may partially mimic the nasally secreted IFN response reported during BRDC, may be beneficial in cattle. PMID- 10476938 TI - Mucosal cytokine therapy: marked antiviral and antitumor activity. AB - Mucosal administration of the Th1 stimulatory cytokines interleukin-2 (IL-2), IL 12, IL-15, IL-18, or granulocyte-macrophage colony-stimulating factor (GM-CSF) induced antiviral activity in mice challenged systemically with a lethal dose of encephalomyocarditis virus (EMCV) similar to that observed following parenteral administration. In contrast, mucosal administration of the Th2 stimulatory cytokines IL-4, IL-5, IL-10, or IL-13 did not affect significantly the survival of EMCV-infected animals. Mucosal administration of IL-2 or IL-12 also exerted a marked antitumor activity in mice inoculated intravenously with Friend erythroleukemia cells. Recombinant IL-2 and IL-18, but none of the other recombinant cytokines tested, induced low levels of IFN in vitro. Polyclonal antibodies to both mouse and human interferon-alpha/beta (IFN-alpha/beta) abrogated the antiviral activity of IL-2 in vivo, even though the anti-human IFN alpha/beta antibody did not neutralize mouse IFN-alpha/beta, and neither antibody bound to IL-2. IL-15 did not exhibit antiviral activity in IFN-alpha/beta R-/- mice, which are deficient in natural killer (NK) cell activity. These results suggest that mucosal Th1 cytokine therapy induces a soluble factor or activates a specific cell population in the lymphoid or epithelial tissue of the oropharyngeal cavity, which potentiates elimination of virus-infected or neoplasic cells systemically. PMID- 10476939 TI - Oral use of interferon-alpha stimulates ISG-15 transcription and production by human buccal epithelial cells. AB - ISG-15 is a 15-kDa protein encoded by an interferon (IFN)-stimulated gene (ISG), which is transcriptionally regulated by IFN-alpha and IFN-beta. Considered as part of the cytokine network, ISG-15 has the potential to amplify the immunomodulatory effects of these IFNs by enhancing IFN-gamma production, natural killer cell proliferation, and lymphokine-alphactivated killer cell cytotoxicity. To understand better the mechanism(s) of action of orally administered IFN-alpha, we have studied the effect of IFN-alpha on ISG-15 gene expression by human buccal epithelial cells (BEC). For in vitro studies, ISG-15 mRNA and protein levels were measured in BEC incubated for 0.5, 2, and 9 h with 100 or 1,000 IU/ml of human lymphoblastoid IFN-alpha. For in vivo studies, ISG-15 mRNA was measured in BEC samples collected at baseline, and 0.5, 2, and 9 h after 5-20 min of oral rinsing with 10 ml of IFN-alpha (1,000 IU/ml). ISG-15 mRNA was measured by reverse transcriptase polymerase chain reaction (RT-PCR), and ISG-15 protein production by Western Blot analysis. IFN-alpha augmented BEC ISG-15 gene expression in a concentration dependent manner both in vivo and in vitro. We conclude that orally administered IFN-alpha exerts its immunomodulatory effects in humans in part by upregulating the production of ISG-15 by BEC, thereby enhancing the immune reactivity of mucosa-associated lymphocytes. PMID- 10476940 TI - Interferon-alpha upregulates gene expression of aquaporin-5 in human parotid glands. AB - Aquaporins are a family of homologous membrane proteins that function as highly selective water channels. Aquaporin-5 (AQP5) is uniquely present in lacrimal and salivary glands, where it accounts for normal tear and saliva production. We tested the hypothesis that orally administered human interferon-alpha (HuIFN alpha) benefits persons with xerostomia by augmenting the production of AQP5 protein by parotid gland epithelium. Cells from three human parotid glands were cultured with and without human lymphoblastoid IFN-alpha, and assayed for AQP5 mRNA levels by reverse transcriptase polymerase chain reaction (RT-PCR), and AQP5 protein levels by Western blot. Intracellular localization of AQP5 protein was done using confocal microscopy. The functional integrity of the glandular tissue was confirmed by RT-PCR analysis of alpha-amylase 1 and basic proline-rich protein transcripts. AQP5 was constitutively expressed in human parotid gland tissue, with AQP5 protein restricted to the plasma membranes and cytoplasmic vesicles of acinar cells. IFN-alpha augmented AQP5 transcription and protein production in a concentration-dependent manner, and increased the size of intensity of staining of AQP5-containing cytoplasmic vesicles in acinar cells. We conclude that IFN-alpha upregulates AQP5 gene expression in human parotid acinar cells in vitro. To our knowledge, this is the first demonstration that IFN-alpha regulates the gene expression of an aquaporin. PMID- 10476941 TI - Low-dose oral use of human interferon-alpha in cancer patients. AB - In a double-blind placebo-controlled trial, 57 adult subjects with disseminated malignancies were given orally low doses of recombinant human interferon-alpha (rHuIFN-alpha) at 0.05 IU, 0.5 IU, or 5.0 IU/kg body weight. The objective was to determine the efficacy of orally administered rHuIFN-alpha on appetite stimulation and/or weight loss prevention in anorectic cancer patients. Almost two-thirds (64%) of the subjects given 5.0 IU/kg reported an increase in appetite or body weight after 5 weeks in contrast to only 29% of the placebo-treated subjects. However, at the end of the 91-day trial, no significant differences in appetite enhancement or weight gain were noted between these two groups. Additionally, the 5.0 IU/kg treated group experienced half as many deaths as the control group by the conclusion of this 91-day trial. PMID- 10476942 TI - Treatment of primary Sjogren's syndrome with low-dose natural human interferon alpha administered by the oral mucosal route: a phase II clinical trial. IFN Protocol Study Group. AB - The purpose of this investigation was to examine the safety and efficacy of four dosages of natural human interferon-alpha (nHuIFN-alpha) delivered over a 12-week period orally in lozenges (150 IU and 450 IU, once [QD] or three times [TID] daily) compared to placebo in subjects with primary Sjogren's syndrome. This randomized, double-blinded clinical trial demonstrated that nHuIFN-alpha at a dose of 150 IU administered TID by oral lozenge significantly improved stimulated whole saliva output compared to placebo after 12 weeks of treatment. The 150 IU TID dose also was suggestive of benefit for 5 of 7 subjective measures of oral and ocular comfort. IFN lozenges demonstrated a good safety profile, with no serious adverse events found in any treatment group. There were no significant differences between the placebo and the four doses of IFN for adverse events by total number, organ system, severity, dropouts, and number judged to be related to treatment. In conclusion, these results demonstrated that the use of 150 IU IFN lozenges TID for 12 weeks in subjects with primary Sjogren's syndrome improved salivary output and decreased complaints of xerostomia without causing significant adverse medical events. PMID- 10476943 TI - Oral administration of interferon-alpha induces a transient decline in oral mucosal immunoglobulins and an increase in interleukin-5. AB - Although administration of interferon-alpha (IFN-alpha) via the oral-mucosal route has shown efficacy in a variety of human and animal diseases, the mechanism of action of orally administered IFN is not clearly understood. To assess the possibility that IFN-alpha given via a lozenge alters the local mucosal immune system, immunoglobulins (Ig) and cytokines were measured in salivary secretions. Volunteers were given low doses of IFN-alpha and saliva was collected over a 24-h period. IgA and precursor IgM were measured by sandwich enzyme-linked immunosorbent assay (ELISA). Salivary concentrations of interleukin-5 (IL-5), the T helper cytokine primarily responsible for the switch from IgM to IgA, were also determined. After oral administration of IFN-alpha, there was an initial decline in IgM and IgA followed by a return to baseline levels by 8-24 h. This change in Ig concentration was associated with a gradual increase in IL-5, consistent with the return of Ig to baseline as a result of modulation by Ig-mediating cytokines. PMID- 10476944 TI - Reduction of morning stiffness and improvement in physical function in fibromyalgia syndrome patients treated sublingually with low doses of human interferon-alpha. AB - One hundred and twelve fibromyalgia syndrome (FMS) patients were randomized into one of four demographically similar groups (n = 28/group). Sequential primary FMS patient volunteers were to receive daily sublingual placebo or interferon-alpha (IFN-alpha) at 15, 50, or 150 IU. After a screening evaluation, analgesic or sedative hypnotic medications were withdrawn. Two weeks later, daily IFN-alpha or placebo was initiated with follow-up evaluations at 2-week intervals ending with week 6. One primary, three secondary, and seven tertiary variables were assessed. Study outcome was based on improvement in the tender point index (TPI). The TPI did not improve with any IFN-alpha dose. However, significant improvement was seen in morning stiffness and in physical function with the 50 IU IFN-alpha (p < 0.01). None of the other outcome means changed significantly and no adverse events were attributable to IFN-alpha therapy. PMID- 10476945 TI - Lymphocyte markers and natural killer cell activity in fibromyalgia syndrome: effects of low-dose, sublingual use of human interferon-alpha. AB - A clinical study was designed to utilize flow cytometric immunophenotyping and chromium release from cultured tumor target cells to characterize peripheral blood mononuclear leukocyte (PBML) subpopulations and natural killer activity in healthy normal controls (n = 18) and in patients with fibromyalgia syndrome (FMS) at baseline (n = 124) and again after 6 weeks of treatment with low-doses of orally administered human interferon-alpha (IFN-alpha). Volunteer subjects discontinued all analgesic and sedative hypnotic medications for 2 weeks prior to the baseline phlebotomy. Laboratory measures included a complete blood count; a phenotypic analysis of PBML by flow cytometry; and in vitro natural killer (NK) cell activity. After baseline blood sample collection, the FMS patients were randomized to one of four parallel treatment groups (n = 28/group) to receive sublingual IFN-alpha (15 IU, 50 IU, 150 IU), or placebo every morning for 6 weeks. The tests were repeated at week 6 to evaluate treatment effects. At baseline, FMS patients exhibited fewer lymphocytes and more CD25+ T lymphocytes than did normal controls. By week 6, the main significant and consistent change was a decrease in the HLA-DR+ CD4+ subpopulation in the 15 IU and 150 IU treatment groups. These data do not support an immunologically dysfunctional PBML phenotype among patients with FMS as has been observed in the chronic fatigue syndrome. PMID- 10476946 TI - Wellcome Trust loses battle to expand its genome campus. PMID- 10476947 TI - A million volunteers join the online search for extraterrestrial life. PMID- 10476948 TI - Leadership wrangle forces Salk Institute restructuring. PMID- 10476949 TI - California initiative to probe transgenics in large animals. PMID- 10476950 TI - Kansas makes a monkey of itself. PMID- 10476951 TI - Alois Alzheimer and the amyloid debate. PMID- 10476952 TI - Mechanics of the ribosome. PMID- 10476953 TI - Cognitive neuroscience. How do you pay attention? PMID- 10476954 TI - Heart disease. Good cholesterol news. PMID- 10476955 TI - Signal transduction. Rhapsody in G proteins. PMID- 10476956 TI - Eladio Vinuela (1937-99) PMID- 10476957 TI - Parasitoid behaviour and Bt plants. PMID- 10476958 TI - Prelude or requiem for the 'Mozart effect'? PMID- 10476959 TI - Prelude or requiem for the 'Mozart effect'? PMID- 10476960 TI - Structure of a bacterial 30S ribosomal subunit at 5.5 A resolution. AB - The 30S ribosomal subunit binds messenger RNA and the anticodon stem-loop of transfer RNA during protein synthesis. A crystallographic analysis of the structure of the subunit from the bacterium Thermus thermophilus is presented. At a resolution of 5.5 A, the phosphate backbone of the ribosomal RNA is visible, as are the alpha-helices of the ribosomal proteins, enabling double-helical regions of RNA to be identified throughout the subunit, all seven of the small-subunit proteins of known crystal structure to be positioned in the electron density map, and the fold of the entire central domain of the small-subunit ribosomal RNA to be determined. PMID- 10476961 TI - Placement of protein and RNA structures into a 5 A-resolution map of the 50S ribosomal subunit. AB - We have calculated at 5.0 A resolution an electron-density map of the large 50S ribosomal subunit from the bacterium Haloarcula marismortui by using phases derived from four heavy-atom derivatives, intercrystal density averaging and density-modification procedures. More than 300 base pairs of A-form RNA duplex have been fitted into this map, as have regions of non-A-form duplex, single stranded segments and tetraloops. The long rods of RNA crisscrossing the subunit arise from the stacking of short, separate double helices, not all of which are A form, and in many places proteins crosslink two or more of these rods. The polypeptide exit channel was marked by tungsten cluster compounds bound in one heavy-atom-derivatized crystal. We have determined the structure of the translation-factor-binding centre by fitting the crystal structures of the ribosomal proteins L6, L11 and L14, the sarcin-ricin loop RNA, and the RNA sequence that binds L11 into the electron density. We can position either elongation factor G or elongation factor Tu complexed with an aminoacylated transfer RNA and GTP onto the factor-binding centre in a manner that is consistent with results from biochemical and electron microscopy studies. PMID- 10476962 TI - Tracking the evolution of insecticide resistance in the mosquito Culex pipiens. AB - The evolution of pesticide resistance provides some of the most striking examples of darwinian evolution occurring over a human life span. Identification of resistance alleles opens an outstanding framework in which to study the evolution of adaptive mutations from the beginning of pesticide application, the evolution of interactions between alleles (dominance) or between loci (epistasis). Here we show that resistance alleles can also be used as markers to dissect population processes at a microevolutionary scale. We have focused on the antagonistic roles of selection and migration involved in the dynamics of local adaptation with reference to allelic frequencies at two resistance loci in the mosquito Culex pipiens. We find that their frequencies follow an annual cycle of large amplitude (25%), and we precisely unravel the seasonal variation of migration and selection underlying this cycle. Our results provide a firm basis on which to devise an insecticide treatment strategy that will better control the evolution of resistance genes and the growth of mosquito populations. PMID- 10476963 TI - Illusory shifts in visual direction accompany adaptation of saccadic eye movements. AB - A central problem in human vision is to explain how the visual world remains stable despite the continual displacements of the retinal image produced by rapid saccadic movements of the eyes. Perceived stability has been attributed to 'efferent-copy' signals, representing the saccadic motor commands, that cancel the effects of saccade-related retinal displacements. Here we show, by means of a perceptual illusion, that traditional cancellation theories cannot explain stability. The perceptual illusion was produced by first inducing adaptive changes in saccadic gain (ratio of saccade size to target eccentricity). Following adaptation, subjects experienced an illusory mislocalization in which widely separated targets flashed before and after saccades appeared to be in the same place. The illusion shows that the perceptual system did not take the adaptive changes into account. Perceptual localization is based on signals representing the size of the initially-intended saccade, not the size of the saccade that is ultimately executed. Signals representing intended saccades initiate a visual comparison process used to maintain perceptual stability across saccades and to generate the oculomotor error signals that ensure saccadic accuracy. PMID- 10476964 TI - Electrophysiological measurement of rapid shifts of attention during visual search. AB - The perception of natural visual scenes that contain many objects poses computational problems that are absent when objects are perceived in isolation. Vision researchers have captured this attribute of real-world perception in the laboratory by using visual search tasks, in which subjects search for a target object in arrays containing varying numbers of non-target distractor objects. Under many conditions, the amount of time required to detect a visual search target increases as the number of objects in the stimulus array increases, and some investigators have proposed that this reflects the serial application of attention to the individual objects in the array. However, other investigators have argued that this pattern of results may instead be due to limitations in the processing capacity of a parallel processing system that identifies multiple objects concurrently. Here we attempt to address this longstanding controversy by using an electrophysiological marker of the moment-by-moment direction of attention-the N2pc component of the event-related potential waveform--to show that attention shifts rapidly among objects during visual search. PMID- 10476965 TI - Global and fine information coded by single neurons in the temporal visual cortex. AB - When we see a person's face, we can easily recognize their species, individual identity and emotional state. How does the brain represent such complex information? A substantial number of neurons in the macaque temporal cortex respond to faces. However, the neuronal mechanisms underlying the processing of complex information are not yet clear. Here we recorded the activity of single neurons in the temporal cortex of macaque monkeys while presenting visual stimuli consisting of geometric shapes, and monkey and human faces with various expressions. Information theory was used to investigate how well the neuronal responses could categorize the stimuli. We found that single neurons conveyed two different scales of facial information in their firing patterns, starting at different latencies. Global information, categorizing stimuli as monkey faces, human faces or shapes, was conveyed in the earliest part of the responses. Fine information about identity or expression was conveyed later, beginning on average 51 ms after global information. We speculate that global information could be used as a 'header' to prepare destination areas for receiving more detailed information. PMID- 10476966 TI - Caudal is the Hox gene that specifies the most posterior Drosophile segment. AB - The homeobox gene caudal (cad) has a maternal embryonic function that establishes the antero-posterior body axis of Drosophila. It also has a conserved late embryonic and imaginal function related to the development of the posterior body region. Here we report the developmental role of cad in adult Drosophila. It is required for the normal development of the analia structures, which derive from the most posterior body segment. In the absence of cad function, the analia develop like the immediately anterior segment (male genitalia), following the transformation rule of the canonical Hox genes. We also show that cad can induce ectopic analia development if expressed in the head or wing. We propose that cad is the Hox gene that determines the development of the fly's most posterior segment. cad acts in combination with the Hedgehog (Hh) pathway to specify the different components of the analia: the activities of cad and of the Hh pathway induce Distal-less expression that, together with cad, promote external analia development. In the absence of the Hh pathway, cad induces internal analia development, probably by activating the brachyenteron and even-skipped genes. PMID- 10476967 TI - Notch signalling controls pancreatic cell differentiation. AB - The pancreas contains both exocrine and endocrine cells, but the molecular mechanisms controlling the differentiation of these cell types are largely unknown. Despite their endodermal origin, pancreatic endocrine cells share several molecular characteristics with neurons, and, like neurons in the central nervous system, differentiating endocrine cells in the pancreas appear in a scattered fashion within a field of progenitor cells. This indicates that they may be generated by lateral specification through Notch signalling. Here, to test this idea, we analysed pancreas development in mice genetically altered at several steps in the Notch signalling pathway. Mice deficient for Delta-like gene 1 (Dll1) or the intracellular mediator RBP-Jkappa showed accelerated differentiation of pancreatic endocrine cells. A similar phenotype was observed in mice over-expressing neurogenin 3 (ngn 3) or the intracellular form of Notch3 (a repressor of Notch signalling). These data provide evidence that ngn3 acts as proendocrine gene and that Notch signalling is critical for the decision between the endocrine and progenitor/exocrine fates in the developing pancreas. PMID- 10476968 TI - A C. elegans Ror receptor tyrosine kinase regulates cell motility and asymmetric cell division. AB - Ror kinases are a family of orphan receptors with tyrosine kinase activity that are related to muscle specific kinase (MuSK), a receptor tyrosine kinase that assembles acetylcholine receptors at the neuromuscular junction. Although the functions of Ror kinases are unknown, similarities between Ror and MuSK kinases have led to speculation that Ror kinases regulate synaptic development. Here we show that the Caenorhabditis elegans gene cam-1 encodes a member of the Ror kinase family that guides migrating cells and orients the polarity of asymmetric cell divisions and axon outgrowth. We find that tyrosine kinase activity is required for some of the functions of CAM-1, but not for its role in cell migration. CAM-1 is expressed in cells that require its function, and acts cell autonomously in migrating neurons. Overexpression and loss of cam-1 function result in reciprocal cell-migration phenotypes, indicating that levels of CAM-1 influence the final positions of migrating cells. Our results raise the possibility that Ror kinases regulate cell motility and asymmetric cell division in organisms as diverse as nematodes and mammals. PMID- 10476969 TI - Bid-deficient mice are resistant to Fas-induced hepatocellular apoptosis. AB - The protein Bid is a participant in the pathway that leads to cell death (apoptosis), mediating the release of cytochrome c from mitochondria in response to signals from 'death' receptors known as TNFR1/Fas on the cell surface. It is a member of the proapoptotic Bcd-2 family and is activated as a result of its cleavage by caspase 8, one of a family of proteolytic cell-death proteins. To investigate the role of Bid in vivo, we have generated mice deficient for Bid. We find that when these mice are injected with an antibody directed against Fas, they nearly all survive, whereas wild-type mice die from hepatocellular apoptosis and haemorrhagic necrosis. About half of the Bid-deficient animals had no apparent liver injury and showed no evidence of activation of the effector caspases 3 and 7, although the initiator caspase 8 had been activated. Other Bid deficient mice survived with only moderate damage: all three caspases (8 and 37) were activated but their cell nuclei were intact and no mitochondrial cytochrome c was released. We also investigated the effects of Bid deficiency in cultured cells treated with anti-Fas antibody (hepatocytes and thymocytes) or with TNFalpha. (fibroblasts). In these Bid-/- cells, mitochondrial dysfunction was delayed, cytochrome c was not released, effector caspase activity was reduced and the cleavage of apoptosis substrates was altered. This loss-of-function model indicates that Bid is a critical substrate in vivo for signalling by death receptor agonists, which mediates a mitochondrial amplification loop that is essential for the apoptosis of selected cells. PMID- 10476970 TI - Activation of the ERK/MAPK pathway by an isoform of rap1GAP associated with G alpha(i) AB - Many receptors for neuropeptides and hormones are coupled with the heterotrimeric G(i) protein, which activates the p42/44 mitogen-activated protein kinase (ERK/MAPK) cascade through both the alpha- and betagamma-subunits of G(i). The betagamma-subunit activates the ERK/MAPK cascade through tyrosine kinase. Constitutively active G(alpha)i2 (gip2) isolated from adrenal and ovarian tumours transforms Rat-1 fibroblasts and also activates the ERK/MAPK cascade by an unknown mechanism. The ERK/MAPK pathway is activated by Ras, and is inhibited when the low-molecular-mass GTP-binding protein Rap1 antagonizes Ras function. Here we show that a novel isoform of Rapl GTPase-activating protein, called rap1GAPII, binds specifically to the alpha-subunits of the G(i) family of heterotrimeric G-proteins. Stimulation of the G(i)-coupled m2-muscarinic receptor translocates rap1GAPII from the cytosol to the membrane and decreases the amount of GTP-bound Rap1. This decrease in GTP-bound Rap1 activates ERK/MAPK. Thus, the alpha-subunit of G(i) activates the Ras-ERK/MAPK mitogenic pathway by membrane recruitment of rap1GAPII and reduction of GTP-bound Rap1. PMID- 10476971 TI - Extracellular sodium regulates airway ciliary motility by inhibiting a P2X receptor. AB - The mucociliary system is responsible for clearing inhaled particles and pathogens from the airways. This important task is performed by the beating of cilia and the consequent movement of mucus from the lungs to the upper airways. Because ciliary motility is enhanced by elevated intracellular calcium concentrations, inhibition of calcium influx could lead to disease by jeopardizing mucociliary clearance. Several hormones and neurotransmitters stimulate ciliary motility, one of the most potent of which is extracellular ATP (ATP0), which acts by releasing calcium ions from internal stores and by activating calcium influx. Here we show that, in airway ciliated cells, extracellular sodium ions (Na+(0)) specifically and competitively inhibit an ATP0 gated channel that is permeable to calcium ions, and thereby attenuate ATP0 induced ciliary motility. Our finding points to a physiological role for Na+(0) in ciliary function, and indicates that mucociliary clearance might be improved in respiratory disorders such as chronic bronchitis and cystic fibrosis by decreasing the sodium concentration of the airway surface fluid in which the cilia are bathed. PMID- 10476972 TI - A new pathway for polyketide synthesis in microorganisms. AB - Chalcone synthases, which biosynthesize chalcones (the starting materials for many flavonoids), have been believed to be specific to plants. However, the rppA gene from the Gram-positive, soil-living filamentous bacterium Streptomyces griseus encodes a 372-amino-acid protein that shows significant similarity to chalcone synthases. Several rppA-like genes are known, but their functions and catalytic properties have not been described. Here we show that a homodimer of RppA catalyses polyketide synthesis: it selects malonyl-coenzyme-A as the starter, carries out four successive extensions and releases the resulting pentaketide to cyclize to 1,3,6,8-tetrahydroxynaphthalene (THN). Site-directed mutagenesis revealed that, as in other chalcone synthases, a cysteine residue is essential for enzyme activity. Disruption of the chromosomal rppA gene in S. griseus abolished melanin production in hyphae, resulting in 'albino' mycelium. THN was readily oxidized to form 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin), which then randomly polymerized to form various coloured compounds. THN formed by RppA appears to be an intermediate in the biosynthetic pathways for not only melanins but also various secondary metabolites containing a naphthoquinone ring. Therefore, RppA is a chalcone-synthase-related synthase that synthesizes polyketides and is found in the Streptomyces and other bacteria. PMID- 10476973 TI - Mastery of fundamental motor skills among New South Wales school students: prevalence and sociodemographic distribution. AB - Mastery of fundamental motor skills among children and adolescents is a potentially important contribution to satisfying participation in sports, games and other physical activities and may enhance the development of an active lifestyle. However, few attempts have been made to determine the prevalence of fundamental motor skill mastery among young Australians. The NSW Schools Fitness and Physical Activity Survey, 1997 (N = 5518) randomly selected schools proportionally from all three education sectors and selected students in Years 4, 6, 8 and 10. Performance on six fundamental motor skills (run, vertical jump, catch, overhand throw, forehand strike and kick) was assessed qualitatively. The prevalence of mastery and near mastery of each skill and mastery of each skill component is reported for boys and girls in each school year. The findings indicate that the prevalence of mastery and near mastery of each of the fundamental motor skills was generally low. There were no differences between students from urban or rural schools and the prevalence of skill mastery was directly associated with socioeconomic status more consistently among girls than among boys. Greater curriculum time and resourcing and training of teachers is required to increase the proportion of students who have mastered the skills fundamental to common sports, games and other physical activities. PMID- 10476974 TI - The influence of plyometric training on the freestyle tumble turn. AB - This study examined the effects of a plyometric training program on freestyle tumble turns. Thirty-eight age group swimmers were assigned to a control group which swam 1.5 hours, three times per week for 20 weeks; or an experimental group which supplemented 1.25 hours of swimming with 15 minutes of plyometrics for the same time frame. The same coach conducted all swimming and plyometric sessions to ensure uniformity. Swimming performance was assessed from 50 m time. Freestyle turning performance was measured by 2.5 m round trip time (RTT), 5 m RTT, wall contact time and selected kinematic and kinetic variables associated with the turn. A Plyopower system was also used to test jump height and velocity. Repeated measures, multivariate analysis of variance showed no significant differences between the groups (pre-, mid- and post-intervention) over the period of the study for any swimming, kinetic or plyopower measures. Thus, equal benefits were derived from normal practice time in the water or land based plyometric exercises. PMID- 10476975 TI - Athletic injury and minor life events: a prospective study. AB - Minor life events were examined in an attempt to determine their contribution to athletic injury risk. Male and female athletes (N = 98), from both team and individual sports (hockey, volleyball, and triathlon) were examined over the course of a competitive season. A high rate of injury was noted among these athletes (30%-46% of the members of each team sustained at least one injury during the season). The injured athletes were found to have a significant increase in minor life events for the week prior to injury. No significant changes in minor life events occurred for the non-injured athletes. The results of the present study provide substantial evidence for a link between minor life events and athletic injury. PMID- 10476976 TI - The use of physical fitness scores and anthropometric data to predict selection in an elite under 18 Australian rules football team. AB - This study was conducted to determine if anthropometric and fitness testing scores can be used to discriminate between players that were selected or not selected in an elite Under 18 Australian Rules Football side. A training squad of 40 Australian Rules Football players was assessed on a battery of standard anthropometric and fitness tests just prior to the selection of the 30 man player roster for the upcoming season. Results showed that the selected players were significantly (P<0.05) taller and had greater upper body strength than non selected players. A discriminant analysis was performed which predicted with an accuracy of 80% whether each player was successful or unsuccessful in gaining selection. This suggested that physical conditioning and anthropometric measurements do play an important part in determining selection in elite junior Australian Rules Football teams. However the discriminant function predicted non selected players (90.9%) better than it predicted selected players (75.9%). Selected Under 18 players were found to be similar to the values reported for elite to sub-elite senior players on height, sit and reach, CMJ and perhaps aerobic fitness, but considerably less than the senior players on 3RM bench press and body mass. PMID- 10476977 TI - An analysis of movement and discomfort of the female breast during exercise and the effects of breast support in three cases. AB - Breast pain is common during exercise, occurring in up to 56% of subjects in some surveys. This pain is mainly associated with the movement of breast tissue. In an attempt to analyse this movement and the resulting pain, the movement of the female breast tissue was quantified in four conditions of breast support ("sports bra", "fashion bra", "crop top" and bare breasted) during four different activities (running, jogging, aerobics march and walking). These activities represented general patterns of exercise for adult females in Australian society. All three subjects were healthy, active, young women of varying breast size (12B, 14B & 14C) that were typical of young women. Two of the subjects had noted breast pain whilst exercising. The results showed that wearing external support for the breast tissue reduced absolute vertical movement and maximum downward deceleration force on the breast. Support also reduced perceived pain. When compared to other forms of breast support a "sports bra" (brassiere designed for breast support whilst exercising) provided superior support for the breast in relation to the amplitude of movement, deceleration forces on the breast, and perceived pain. The data indicates that adult females should wear appropriate breast support to reduce perceived breast discomfort or pain. Of the three garments examined in this study, the fitted sports bra provided superior support and pain reduction. PMID- 10476978 TI - The development of a locus of control measure predictive of injured athletes' adherence to treatment. AB - The Sports Rehabilitation Locus of Control (SRLC) scale was developed to facilitate study of the behaviour of athletes recovering from injury. The SRLC was built specifically to assess locus of control in injured athletes by adapting selected items from the Multidimensional Health Locus of Control (MHLC) questionnaire developed by Wallston, Wallston, & DeVellis (1978), and in accordance with their rationale for the construction of domain-specific LOC scales. The resultant locus of control measures (internal, powerful others, and chance) were administered (along with the MHLC) to a sample of 145 sports-injured athletes. Good internal consistency (alphas ranged from .72 to .79) and test retest reliability (correlations ranged from .75 to .85) was shown by the SRLC scales, and the pattern of inter-scale correlations supported the three-factor conceptualisation of locus of control on which the SRLC was based. In the subsequent initial validation study, the relationship between SRLC scale scores and adherence to treatment was tested with a sample of injured athletes. Encouragingly, internality was positively associated p<.05 with treatment adherence. PMID- 10476979 TI - The nature and circumstances of tackle injuries in rugby union. AB - This study describes the nature and circumstances of injury occurring in rugby union tackles (33% of 569 injury events) using data from the Rugby Injury and Performance Project (RIPP) and provides supplementary information on the nature of tackles involving injury from analysis of videotape of tackle injury events. The most common tackle injuries in the RIPP data were sprains/strains (41%) followed by haematomas/bruising (26%). The most frequently injured body sites were the head/neck/face (22%) and the knee (17%). The ball carrier and tackler were injured in tackles in similar proportions in both RIPP and New Zealand Rugby Football Union (NZRFU) video tackle incidents. Both players were most often in motion in the tackle at the time of injury with approximately 70% of injuries occurring when the injured player was running or diving/falling to the ground. Tackle injury was most often caused by impact with another player rather than impact with the ground. The use of protective padding may reduce the risk of impact injury. The majority of tackle injuries were associated with stopping tackles to the trunk which were from the front (63%), rather than from the side or behind. Thus consideration should be given to coaching strategies or to rule changes which reduce the likelihood or prohibit front-on tackles. PMID- 10476980 TI - The future of public health. PMID- 10476981 TI - NIDCR combats oral complications of cancer treatment. PMID- 10476982 TI - Agencies target diabetes. PMID- 10476984 TI - Women's health exhibit to tour US. PMID- 10476983 TI - Health education guides target Hispanic Americans. PMID- 10476985 TI - More research needed to ensure environmental justice. PMID- 10476986 TI - WHO launches Vision 2020 to combat avoidable blindness. PMID- 10476987 TI - WHO tags 1999-2000 vaccine, prepares flu pandemic plan. PMID- 10476988 TI - WHO pushes first cervical cancer vaccine. PMID- 10476989 TI - Building the next generation of healthy people. PMID- 10476990 TI - HRSA's Models That Work Program: implications for improving access to primary health care. AB - The main objective of the Models That Work Campaign (MTW) is improving access to health care for vulnerable and underserved populations. A collaboration between the Bureau of Primary Health Care (BPHC) at the Health Resources and Services Administration (HRSA) and 39 cosponsors--among them national associations, state and federal agencies, community-based organizations, foundations, and businesses- this initiative gives recognition and visibility to innovative and effective service delivery models. Models are selected based on a set of criteria that includes delivery of high quality primary care services, community participation, integration of health and social services, quantifiable outcomes, and replicability. Winners of the competition are showcased nationally and hired to provide training to other communities, to document and publish their strategies, and to provide onsite technical assistance on request. PMID- 10476991 TI - Medicaid managed care and public health data. AB - In summary, there are a number of ways in which state public health data can be of value in the design of Medicaid managed care plans. At the level of the purchaser, such as a state Medicaid agency, public health data can assist in decision-making around pricing policy and can be useful in prioritizing interventions for those conditions that most severely affect the covered population. Quality assurance standards such as the HEDIS clinical performance measures can be used to define a baseline of prevention-oriented services or, by adding additional customized data points, to emphasize a particular service. From the standpoint of the managed care plan, public health data can be useful in understanding the needs of a community it serves or would like to serve and in estimating the prevalence of various conditions in that community that will influence the premium it will charge. Thus, there are multiple routes through which public health goals and priorities can be incorporated into managed care and can leverage the power of managed care to improve the public's health. PMID- 10476992 TI - Sharing responsibility for the public's health. PMID- 10476993 TI - Development of a surveillance program for occupational pesticide poisoning: lessons learned and future directions. AB - The authors describe the growth from 1987 through 1996 of the Occupational Pesticide Poisoning Surveillance Program at the Texas Department of Health. The program was initially based on a Sentinel Event Notification System for Occupational Risks (SENSOR) model, using sentinel providers to report cases, supplementing the passive reporting by physicians that was required by law. The model was evaluated after five years, and significant changes were implemented to improve case ascertainment. Current active surveillance methods emphasize collaboration with a number of agencies and organizations for identification of cases and follow-up. The number of confirmed occupational cases increased from 9 workers in 1987 to 99 workers in 1996. The evolution from a passive system to an active surveillance program expanded the number of reported cases and strengthened inter-agency collaborations. PMID- 10476994 TI - Viral meningitis in child care center staff and parents: an outbreak of echovirus 30 infections. AB - OBJECTIVE: A report of five cases of viral meningitis among adults with children enrolled in a child care center prompted an investigation of risk factors for viral transmission from children to adult household members. METHODS: To determine recent echovirus 30 (E30) infections, the authors conducted a serologic survey. To determine risk factors for infection among adult household members, they conducted a retrospective cohort study using written questionnaires. RESULTS: Recent E30 infections were found in 84% of children tested, 57% of adult household members tested, and 47% of staff members tested. Infected adults were more likely than infected children to have clinical meningitis. Among adult household members, changing diapers was a risk factor for recent infection. Women who changed > or = 90 diapers per month had a higher infection rate than women who changed fewer diapers; in contrast, men who changed > or = 90 diapers per month had a lower infection rate than men who changed fewer diapers. Handwashing was protective: there was a negative correlation between handwashing after diaper changes and E30 infection among adults with infected children in diapers. CONCLUSIONS: Because child care centers can be a source of enteroviral infections among adult household members, adults with viral meningitis should be questioned about their children's day care or preschool attendance. The importance of handwashing should be stressed to adults with children in day care. PMID- 10476995 TI - Asthma prevalence among American Indian and Alaska Native children. AB - OBJECTIVES: Although asthma is the most common chronic childhood illness in the United States, little is known about its prevalence among American Indian and Alaska Native (AI/AN) children. The authors used the latest available household survey data to estimate the prevalence of asthma in this population. METHODS: The authors analyzed data for children ages 1 through 17 years from the 1987 Survey of American Indians and Alaska Natives (SAIAN) and the 1987 National Medical Expenditure Survey (NMES). At least one member of each AI/AN household included in the SAIAN was eligible for services through the Indian Health Service. RESULTS: The weighted prevalence of parent-reported asthma was 7.06% among 2288 AI/AN children ages 1-17 (95% CI 5.08, 9.04), compared with a US estimate of 8.40% for children ages 1-17 based on the 1987 NMES (95% CI 7.65, 9.15). The AI/AN sample was too small to yield stable estimates for a comparison between AI/AN children and all US children when the data were stratified according to household income and metropolitan vs non-metropolitan residence. The unadjusted asthma prevalence rates were similar for AI/AN children and for children in the NMES sample. CONCLUSIONS: In 1987, the prevalence of parent-reported asthma was similar for AI/AN children in the SAIAN sample and for children in the NMES sample. More recent data are needed to better understand the current prevalence of asthma among AI/AN children. PMID- 10476996 TI - Domestic violence and sexually transmitted diseases: the experience of prenatal care patients. AB - OBJECTIVES: The authors analyzed interview responses of patients at a prenatal care clinic to explore whether women who had been victims of sexual and physical abuse were more likely than non-victimized women to have experienced a sexually transmitted disease (STD). METHODS: A consecutive sample of 774 prenatal patients of a large health department in North Carolina were interviewed concerning a variety of health issues, including violence and STDs. Logistic regression analysis was used to model the women's STD status as a function of their experiences of sexual and physical abuse, controlling for several potentially confounding factors. RESULTS: Thirty percent of the women reported having experienced at least one STD, with the most common infections being chlamydia and gonorrhea. Twenty-eight percent of the women reported having been victims of abuse; 16% reported physical abuse only, while 12% reported both physical and sexual abuse. The majority of violence was domestic in nature, perpetrated by the victims' husbands, boyfriends, male friends, and relatives. After controlling for confounding variables, the authors found that women who reported both physical and sexual abuse were significantly more likely to have experienced STDs than non victims (odds ratio [OR] = 2.25; 95% confidence interval [CI] 1.37, 3.69). The logistic regression analysis also showed a relationship of borderline statistical significance between non-sexual physical abuse and STDs. CONCLUSIONS: Health care providers should routinely screen patients for both abuse and STDs, and they should assist identified women in accessing appropriate health, social, and legal services. PMID- 10476997 TI - Cross-matching TB and AIDS registries: TB patients with HIV co-infection, United States, 1993-1994. AB - OBJECTIVES: Because of limited reporting of HIV status in case reports to the national tuberculosis (TB) surveillance system, the authors conducted this study to estimate the proportion of US TB cases with HIV co-infection and to describe demographic and clinical characteristics of co-infected patients. METHODS: The 50 states, New York City, and Puerto Rico submitted the results of cross-matches of TB registries and HIV-AIDS registries. The authors determined the number of TB cases reported for 1993-1994 that were listed in HIV-AIDS registries and analyzed data on demographic and clinical characteristics by match status. RESULTS: Of 49,938 TB cases reported for 1993-1994, 6863 (14%) were listed in AIDS or HIV registries. The proportions of TB-AIDS cases among TB cases varied by reporting area, from 0% to 31%. Anti-TB drug resistance was higher among TB-AIDS cases, particularly resistance to isoniazid and rifampin (multidrug resistance) and rifampin alone, In some areas with low proportions of multidrug-resistant TB cases, however, the difference in multidrug resistance between TB-AIDS patients and non-AIDS TB patients was not found. CONCLUSIONS: The proportion of TB cases with HIV co-infection, particularly in some areas, underscores the importance of the HIV-AIDS epidemic for the epidemiology of TB. Efforts to improve HIV testing as well as reporting of HIV status for TB patients should continue to ensure optimum management of coinfected patients, enhance surveillance activities, and promote judicious resource allocation and targeted prevention and control activities. PMID- 10476998 TI - Extreme exposure: the Sydney to Hobart yacht race and the role of prediction. PMID- 10476999 TI - The Initiative to Eliminate Racial and Ethnic Health Disparities is moving forward. PMID- 10477000 TI - Results of proton therapy of uveal melanomas treated in Nice. AB - PURPOSE: To present the first results of uveal melanomas treated with the Medicyc Cyclotron 65 MeV proton beam facility in Nice, analyzing the factors that affect the cause-specific survival (CSS), metastatic rate, and reporting the visual outcome. METHODS AND MATERIALS: This study concerns 538 patients referred by French institutions between June 1991 and December 1996. The eye and tumor parameters were measured using ultrasonography and angiography. Since 1994, CT scans were performed in most patients to help determine the axial length and the shape of the ocular globe. Tantalum clips were inserted around the tumor by the referring ophthalmologist. There were 349 posterior pole tumors (64.9%), 130 equatorial tumors (24.1%), and 59 ciliary body tumors (11%). Two hundred four patients (37.9%) had T1 or T2 tumors, and 334 patients (62.1%) had T3 or T4 tumors. The median tumor diameter was 14.6 mm, and the median tumor height was 5.1 mm. All patients received 52 Gy (57.20 Gy Co-equivalent dose) on 4 consecutive days. The data were analyzed by December 1997. RESULTS: The CSS was 77.4% at 78 months, the overall survival was 73.8% and the local control was 89.0%. The CSS was not influenced by the patient age or the site of the tumor. It was 81.5% for T1 and T2 tumors, versus 75% for T3 and T4 tumors (P = 0.035). It was found that the tumor diameter, rather than the height, was the most important parameter affecting outcome. The metastatic rate was 8%. It depended on the T stage, tumor diameter and thickness, but not the tumor site. Thirty-eight enucleations were performed, most of them due to tumor progression and/or glaucoma. One-third of the patients in whom visual acuity was adequately scored before and after treatment had a stable, if not improved vision, and half the patients retained useful vision after treatment. CONCLUSION: The outcome of patients suffering from uveal melanoma and treated with high-energy protons compares favorably with other techniques of treatment. The tumor dimensions affected CSS and metastatic rate. Even though two-thirds of patients had posterior pole tumors, half of them retained useful vision. PMID- 10477001 TI - Carcinoma of the larynx treated with hypofractionated radiation and hyperbaric oxygen: long-term tumor control and complications. AB - PURPOSE: To evaluate the long-term outcome with respect to local control, survival, and complications in a cohort of patients with locally advanced laryngeal carcinoma treated with hypofractionated radiation and hyperbaric oxygen at 4 atmospheres of pressure (HBO-4). METHODS AND MATERIALS: Between January 1970 and August 1982, 45 patients with locally advanced carcinoma of the larynx were treated with primary radiation using a unique hypofractionated schedule of 2 fractions of 11 Gy separated by 21 days, with concomitant HBO-4 during each radiotherapy session. To avoid seizures, discomfort and other complications of HBO-4, each session was performed under general anesthesia. All patients had pathologically confirmed squamous cell carcinoma of the glottic (23) or supraglottic larynx (22) and were staged as follows: T2-5, T3-24, T-4-16; N0-26, N1-4, N2-13, N3-1. Patients were treated with opposed lateral wedged fields of 4 6 MV photons, with a median field size of 5.5 x 9.75 to a total median dose of 22.5 Gy. RESULTS: As of February 1998, follow-up was complete on all but one patient, who relocated to another country after 8 years. Complete clinical responses were observed in 39 (87%) of the cases. The 10-year local control rate for all 45 patients was 58%, and local control for the complete responders was 69%. Three patients underwent laryngectomy for complications and were found to have no pathological evidence of disease in the laryngectomy specimen. The 10 year survival of the overall population was 27%. The 10-year voice preservation rate for the the 39 complete responders was 55%. Acute mucosal and skin reactions were modest and acceptable. Significant late complications occurred in 14 patients consisting of severe fibrosis, necrosis, pharyngeal fistula, with 3 patients requiring laryngectomy for complications. The actuarial rate of severe complications at 5 years was 42%. CONCLUSIONS: The response rate and long-term tumor control rate obtained with this treatment program were comparable to more protracted radiation schedules with or without systemic chemotherapy. The complication rate was high resulting in an adverse therapeutic ratio. The radiobiologic interpretation of this clinical data, and implications for hypoxia directed therapy, are discussed. PMID- 10477002 TI - Smart (simultaneous modulated accelerated radiation therapy) boost: a new accelerated fractionation schedule for the treatment of head and neck cancer with intensity modulated radiotherapy. AB - PURPOSE: To report the initial experience in the definitive treatment of head and neck carcinomas using SMART (Simultaneous Modulated Accelerated Radiation Therapy) boost technique. Radiation was delivered via IMRT (Intensity Modulated Radiotherapy). The following parameters were evaluated: acute toxicity, initial tumor response, clinical feasibility, dosimetry and cost. METHODS AND MATERIALS: Between January 1996 and December 1997, 20 patients with primary head and neck carcinomas were treated with SMART boost technique. The treatment fields encompassed two simultaneous targets. The primary target included palpable and visible disease sites. The secondary target included regions at risk for microscopic disease. Daily fractions of 2.4 Gy and 2 Gy were prescribed and delivered to the primary and secondary targets to a total dose of 60 Gy and 50 Gy, respectively. Lower neck nodes were treated with a single conventional anterior portal. This fractionation schedule was completed in 5 weeks with 5 daily fractions weekly. Toxicity was evaluated by RTOG acute toxicity grading criteria, evidence of infection at immobilization screw sites, subjective salivary function, weight loss, and the need for treatment split. Mean follow-up was 15.2 months. Initial tumor response was assessed by clinical and radiographical examinations. Clinical feasibility was evaluated by the criteria: time to treat patient, immobilization, and treatment planning and QA time. In dosimetry, we evaluated the mean doses of both targets and normal tissues and percent targets' volume below goal. To evaluate cost, Medicare allowable charge for SMART boost was compared to those of conventional fractionated and accelerated radiotherapy. RESULTS: ACUTE TOXICITY: None of the patients had a screw site infection and all patients healed well after completion of radiotherapy. Sixteen of 20 patients (80%) completed the treatment within 40 days without any split. Sixteen patients (80%) had RTOG Grade 3 mucositis while 10 patients (50%) had Grade 3 pharyngitis. Three of 20 patients (15%) had weight loss greater than 10% of their pretreatment weight. Ten patients (50%) required intravenous fluids, tube feeding or both. Nine patients (45%) reported moderate xerostomia with significant relief reported within 6 months. INITIAL TUMOR RESPONSE: 19 patients (95 %) had complete response (CR) while one had partial response (PR). The patient with PR had stable disease on imaging at 12 months follow-up. Two patients were found to have lung metastases at 2 months and 5 months follow-up. To date, there have been two local recurrences in the complete responders. Both patients had nasopharyngeal primary; one was retreated with radioactive Cesium-137 implant and the other died from the disease. CLINICAL FEASIBILITY: The average treatment time for a three-arc treatment was 17.5 minutes and 2.5 minutes for each additional arc. Eleven patients (55%) had four arc treatment while six patients (30%) had five-arc treatment and three patients (15%) had three-arc treatment. Immobilization was reproducible within less than 2 mm. The treatment planning, QA and documentation prior to treatment averaged 2 days. DOSIMETRY: The mean doses to the primary and secondary targets were 64.4 Gy and 54.4 Gy, respectively; 8.9% of the primary target volume and 11.6% of the secondary target volume were below prescribed dose goal. The mean dose delivered to the mandible was 30 Gy, spinal cord 17 Gy, ipsilateral parotid 23 Gy, and contralateral parotid 21 Gy. COST: Total Medicare allowable charge for SMART boost was $7000 compared to $8600 (conventional) and $9400 (accelerated fractionation). CONCLUSIONS: SMART boost technique is an accelerated radiotherapy scheme that can be delivered with acceptable toxicity. It allows parotid sparing as evidenced both clinically and by dosimetry. Initial tumor response has been encouraging. It is clinically feasible and cost saving. A larger population of patients and a long-term fol PMID- 10477003 TI - A three-dimensional CT-based target definition for elective irradiation of the neck. AB - INTRODUCTION: Elective treatment of the clinically node-negative neck by radiation results in excellent control rates. However, radiation therapy with its organ-preserving properties is not without morbidity. Side effects of elective neck irradiation are mainly due to damage of the major and minor salivary glands, resulting in the dry mouth syndrome. Given that RT is the preferred treatment modality in case of elective treatment of the neck in many institutions, it is of utmost importance to try and reduce the associated sequelae of RT. MATERIAL AND METHODS: With the introduction of CT-planning systems and the development of 3D conformal radiation therapy (3D CRT) techniques, it has become feasible to deliver adequate doses of radiation to the target (neck) and at the same time saving (parts of) the salivary glands from doses beyond tolerance. A prerequisite for these techniques is that they require a precise knowledge of the target (i.e., of the elective neck) on CT. To be able to correlate borders of the surgical levels in the neck (I-VI) with structures seen on CT, an anatomical study, using two fixed (phenol, formaldehyde) human cadavers, was performed. Subsequently, the 6 potential lymph node regions in the neck on CT were defined. RESULTS AND DISCUSSION: The reference for the current 3D CT-based definition of the lymph node regions in the neck is the official report of the American Academy of Otolaryngology, describing, based on surgical anatomy, the lymph node groups in the neck by Levels I-VI. The present investigation depicts reproducible landmarks on transversal CT images, corresponding to anatomical reference structures known from surgical levels (I-VI) and, this way, CT-based lymph node regions (1-6) were constructed. PMID- 10477004 TI - Avoidance of treatment interruption: an unrecognized benefit of accelerated radiotherapy in oropharyngeal carcinomas? AB - PURPOSE: To assess the impact of treatment interruption on the potential gain in locoregional control obtained with accelerated radiotherapy (RT) compared with conventionally fractionated RT in patients with oropharyngeal carcinomas. METHODS AND MATERIALS: 152 patients treated with radical RT for oropharyngeal carcinomas between 1979 and 1996 were retrospectively analyzed. According to the American Joint Committee on Cancer (AJCC) staging system, there were 6/30/43/73 stages III/III/IV. Sixty-one patients were treated with a conventional RT schedule (median dose 70 Gy in 35 fractions), and 91 patients with either of two 5/5.5 week accelerated RT schedules (median dose 69.6-69.9 Gy in 41 fractions). Discounting weekends, RT was interrupted for 2 consecutive days or more in 53 patients (median duration 11 days, range 2-97), including 67% of the patients in the conventional RT group and 13% in the accelerated RT group. Median follow-up for surviving patients was 55 months (range 23-230). The Cox proportional hazards model was used for the multivariate analysis of factors influencing locoregional control. RESULTS: In univariate analysis, factors associated with a significant decrease in locoregional control included WHO performance status > or =1, advanced AJCC stages (III and IV), conventional RT fractionation, overall treatment time > or =44 days (median), and RT interruption. In the multivariate analysis, when introduced into the model individually, the three significant therapeutic factors remained significant after adjustment for the forced clinical variables. However, when the three therapeutic factors were introduced together into the model, beside the AJCC stage (P = 0.017), only RT interruption remained a significant independent adverse prognostic factor (P = 0.026). CONCLUSIONS: This multivariate analysis highlights the potential negative impact of treatment gaps on locoregional control in oropharyngeal carcinomas. This suggests that treatment interruption may be an even more important parameter than the type of RT schedule per se. Thus, when assessing the relative merit of two RT schedules, inclusion of the other therapeutic factors in a multivariate model is mandatory in order to avoid misinterpretation of the results. PMID- 10477005 TI - Quality of life in patients cured from a carcinoma of the head and neck by radiotherapy: the importance of the target volume. AB - PURPOSE: To assess the health-related quality of life (QOL) of long-term survivors of carcinomas of different subsites of the head and neck following curative radiotherapy (RT). PATIENTS AND METHODS: Patients continuously free from recurrence or second primary tumors treated 1988-1994 were contacted 5.1 to 5.9 years after RT and asked to fill in the EORTC QLQ-C30 core questionnaire and the H&N cancer module. RT had been restricted to the glottis (group A; carcinomas of the vocal cord T1-2 N0), or had included bilateral neck nodes and the primary tumor outside the nasopharynx (group B; AJC Stage II to IV) or within the nasopharynx, respectively (group C; Stage II to IV). Response rate was 97% (group A; n = 41), 69% (group B; n = 26) and 71% (group C; n = 12), respectively. The groups were different with respect to age (older in group A), alcohol consumption (absent in group C) and proportion of females (more in group C). RESULTS: Patients with nasopharyngeal cancer reported the highest morbidity on the H&N module (dry mouth, sticky saliva, trismus, problems with teeth, trouble eating). However, these symptoms did not have a high impact on global QOL or function scores on the QLQ-C30 core questionnaire. Patients in group B reported a lower global QOL but less severe symptoms on the module. CONCLUSION: The high morbidity of patients treated for a nasopharyngeal cancer may be explained by the location of the target volume which included the bilateral temporo-mandibular joints and the salivary glands. These patients require appropriate care during follow-up and will probably profit most from new RT techniques with sparing of normal tissues. PMID- 10477006 TI - The efficacy of early adjuvant radiation therapy for pT3N0 prostate cancer: a matched-pair analysis. AB - PURPOSE: This study examines the effect of adjuvant radiation therapy (RT) on outcome in patients with pT3N0 prostate cancer and makes comparisons to a matched control group. METHODS AND MATERIALS: At our center, 149 patients undergoing radical prostatectomy were found to have pT3N0 prostate cancer, had an undetectable postoperative prostate-specific antigen (PSA) level, and had no immediate hormonal therapy. Fifty-two patients received adjuvant RT within 3 to 6 months of surgery. Ninety-seven underwent radical prostatectomy alone and were observed until PSA failure. From these two cohorts, we matched patients 1:1 according to preoperative PSA (<10 ng/ml vs. >10 ng/ml), Gleason score (<7 vs. > or =7), seminal vesicle invasion, and surgical margin status. Seventy-two patients (36 pairs) were included in the analysis. Median follow-up time was 41 months. We calculated a matched-pairs risk ratio for cumulative risk of PSA relapse (a rise above 0.2 ng/ml). RESULTS: After controlling for the prognostic factors by matching, there was an 88% reduction (95% confidence interval [CI]: 78 93%) in the risk of PSA relapse associated with adjuvant RT. The 5-year freedom from PSA relapse rate was 89% (95% CI: 76-100%) for patients receiving adjuvant RT as compared to 55% (95% CI: 34-79%) for those undergoing radical prostatectomy alone. CONCLUSIONS: These data suggest that adjuvant RT for pT3N0 prostate cancer may significantly reduce the risk of PSA failure as compared to radical prostatectomy alone. Its effect on clinical outcome awaits further follow-up. PMID- 10477007 TI - Urinary morbidity following ultrasound-guided transperineal prostate seed implantation. AB - PURPOSE: To assess the urinary morbidity experienced by patients undergoing ultrasound-guided, permanent transperineal seed implantation for adenocarcinoma of the prostate. METHODS AND MATERIALS: Between September 1992 and September 1997, 693 consecutive patients presented with a diagnosis of clinically localized adenocarcinoma of the prostate, and were treated with ultrasound-guided transperineal interstitial permanent brachytherapy (TPIPB). Ninety-three patients are excluded from this review, having received neoadjuvant antiandrogen therapy. TPIPB was performed with 125I in 165 patients and with 103Pd in 435 patients. Patients treated with implant alone received 160 Gy with 125I (pre TG43) or 120 Gy with 103Pd. One hundred two patients received preimplant, pelvic external beam radiation (XRT) to a dose of either 41.4 or 45 Gy because of high-risk features including PSA > or = 10 and/or Gleason score > or = 7. Combined modality patients received 120 Gy and 90 Gy, respectively for 125I or 103Pd. All patients underwent postimplant cystoscopy and placement of an indwelling Foley catheter for 24-48 h. Follow-up was at 5 weeks after implant, every 3 months for the first 2 years, and then every 6 months for subsequent years. Patients completed AUA urinary symptom scoring questionnaires at initial consultation and at each follow-up visit. Urinary toxicity was classified by the RTOG toxicity scale with the following adaptations; grade 1 urinary toxicity was symptomatic nocturia or frequency requiring none or minimal medical intervention such as phenazopyridine; grade 2 urinary toxicity was early obstructive symptomatology requiring alpha-blocker therapy; and grade 3 toxicity was considered that requiring indwelling catheters or posttreatment transurethral resection of the prostate for symptom relief. Log rank analysis and Chi-square testing was performed to assess AUA score, prostate size, isotope selection, and the addition of XRT as possible prognosticators of postimplant urinary toxicity. The prostate volume receiving 150% of the prescribed dose (V150) was studied in patients to assess its correlation with urinary toxicity. RESULTS: Median follow-up was 37 months (range 6-68). Within the first 60 days, 37.3% of the patients reported grade 1 urinary toxicity, 41% had grade 2, and 2.2% had grade 3 urinary toxicity. By 6 months, 21.4% still reported grade 1 urinary toxicity, whereas 12.8% and 3% complained of grade 2 and 3 urinary difficulties, respectively. Patients with a preimplant AUA score < or = 7 had significantly less grade II toxicity at 60 days compared to those with an AUA score of >7 (32% vs. 59.2%, respectively, p = 0.001). Similarly, prostatic volumes < or = 35 cc had a significantly lower incidence of grade II urinary toxicity (p = 0.001). There was no difference in toxicity regarding the isotope used (p = 0.138 at 60 days, p = 0.45 at 6 months) or the addition of preimplant XRT (p = 0.069 at 60 days, p = 0.84 at 6 months). Twenty-eight patients (4.7%) underwent TURP after 3 isotope half-lives for protracted obstructive symptoms. Five of these men (17%) developed stress incontinence following TURP, but all patients experienced relief of their obstructive symptoms without morbidity at last follow-up. The percent of the prostate receiving 150% of the prescribed dose (V150) did not predict urinary toxicity. CONCLUSIONS: TPIPB is well tolerated but associated with mild to moderate urinary morbidity. Pretreatment prostatic volume and AUA scoring were shown to significantly predict for grade 2 toxicity while the use of preimplant, pelvic XRT and isotope selection did not. Patients undergoing TURP for protracted symptoms following TPIPB did well with a 17% risk of developing stress incontinence. V150 did not help identify patients at risk for urinary morbidity. As transperineal prostate implantation is used more frequently the associated toxicities and the definition of possible pretreatment prognostic factors is necessary to PMID- 10477008 TI - Treatment-related toxicity from a randomized trial of the sequencing of doxorubicin and radiation therapy in patients treated for early stage breast cancer. AB - PURPOSE: There is concern that breast cancer patients treated with left-sided radiation therapy (XRT) and doxorubicin (DOX) may have an increased risk of cardiac toxicity. In addition, the effect of different sequencing of XRT and chemotherapy (CT) on the likelihood of cardiotoxicity, as well as cellulitis, arm edema, or brachial plexopathy, is not well understood. We reviewed the records of patients treated on a randomized trial testing the sequencing of CT and XRT to determine if there was an increase in cardiac events or other complications in patients treated with a total dose of DOX of 180 mg/m2 and XRT, comparing patients with treatment to the left breast and the right breast, and comparing patients treated with initial CT and initial RT. MATERIALS AND METHODS: From June 1984 to December 1992, 244 patients with clinical stage I or II breast cancer were randomized following conservative surgery to receive CT (4 cycles of CAMFP at 3 week intervals) either before or after XRT (45 Gy to the entire breast, followed by a boost of 16 Gy; nodal radiation therapy was optional). Two hundred thirty-one patients were evaluable for the development of cardiac toxicity. The median age at diagnosis was 45 years (range, 20-68). CT doses were: doxorubicin, 45 mg/m2 IV bolus, d 3; methotrexate, 200 mg/m2 IV, d 1 and 15; 5-fluorouracil, 500 mg/m2 IV, d 1; cyclophosphamide, 500 mg/m2 IV, d 1; prednisone 40 mg p.o., d 1-5. A cardiac event was defined as a myocardial infarction or clinical evidence of congestive heart failure. Median follow-up time was 53 months. RESULTS: No cardiac events were observed for patients with either left- or right-sided breast cancer. The sequencing of CT and XRT had no significant effect on the risk of cardiac toxicity, cellulitis, arm edema or brachial plexopathy. CONCLUSIONS: We observed no evidence of an increased risk of cardiac toxicity from the addition of left breast tangential irradiation to DOX at a total dose of 180 mg/m2. Additional follow-up is needed to exclude possible late events. In addition, the sequencing of CT and XRT does not appear to affect the risk of cellulitis, arm edema, or brachial plexopathy. PMID- 10477009 TI - Risk factors for local recurrence after breast-conserving therapy for invasive carcinomas: a case-control study of histological factors and alterations in oncogene expression. AB - PURPOSE: Many studies have focused on histological risk factors for local recurrence (LR) after breast-conserving therapy (BCT). In addition to histological factors, we studied alterations in the expression of various proteins in relation to LR using a case-control approach. METHODS AND MATERIALS: Ninety-nine LR occurred in a patient cohort of 1,481 tumors treated with BCT. These patients were randomly matched, each with two controls. Matching was performed for age group (< or = 50 and > 50 years), pN stage, and follow-up time. Histology slides were reviewed. Immunohistochemical staining was performed for the following proteins: bcl-2, CD31, cyclin D1, E-cadherin, EGF receptor, ER, PR, Ki-67, c-erbB2/neu, and p53. Statistical analyses were performed using conditional logistic regression. RESULTS: Sixty-six cases and 139 controls with invasive carcinoma remained for analysis. The following variables were significant risk factors for LR: young age (p = 0.006), high nuclear grade (p = 0.04), high mitotic count (p = 0.03), extensive DCIS around the tumor (p = 0.02) but not within the tumor, poorly differentiated type of DCIS (p = 0.03), > 20% ki 67 positive cells (p = 0.006), and PR negativity (p = 0.03). When the analysis was performed for patients < or = and > 50 years, these risk factors were found in the older patients, but not in the younger patients. CONCLUSION: High mitotic count and Ki-67 positivity are risk factors for LR. EDCIS surrounding the invasive tumor is a risk factor for LR, especially when of poorly differentiated type. Age is an important risk factor for LR independent of other risk factors, including alterations in oncogene expression. PMID- 10477010 TI - High-dose-rate brachytherapy for early breast cancer: an ambulatory technique. AB - PURPOSE: To evaluate tolerance and efficiency of a boost dose delivered by high dose-rate brachytherapy (HDRB) in conservative treatment of breast cancer. To evaluate the feasibility of brachytherapy on an out-patient basis. METHODS: One hundred and six patients with T1-T2, N0-N1 breast cancers (108 breasts) have been treated with lumpectomy, external irradiation (45 Gy in 5 weeks), and a boost dose on the tumor bed with HDR iridium brachytherapy. Two fractions of 5 Gy were delivered 6 or 24 hours apart. Implantation was done during the lumpectomy (group A: 24 cases) or 3 to 4 weeks after the end of external irradiation (group B: 84 cases). For group B, the application was performed on local anesthesia, and did not require hospitalization. Characteristics of the population were as follows: T1: 77 (71.3%); T2: 31 (28.7%); median tumor size: 1.5 cm; histology: intraductal carcinomas (DCIS): 14 (13%); infiltrative ductal carcinomas (IDC): 84 (77.8%); others: 10 (9.2%). For IDC, surgical margins were found positive in 15 cases, and an extensive intraductal component was present in 22 cases. RESULTS: All ambulatory HDR implants were performed as planned. No immediate toxicity was noticed, except 5 local hematomas. With a median follow-up of 45 months, 5 local relapses were observed (5-year local relapse rate: 5.1%). Only histological grade III was significantly correlated with local relapse. The 5-year disease-free survival and overall survival were respectively 93.8% and 93.3%. Cosmetic result was evaluated in 87 cases, and was good or excellent in 48 cases (63.2%), acceptable in 27 cases, and poor in 5 cases. CONCLUSION: HDRB allows the boost dose to be performed on an out-patient basis. It seems to offer the same local control as other boost techniques for localized breast cancer with acceptable cosmetic results. PMID- 10477011 TI - Patterns of failure and overall survival in patients with completely resected T3 N0 M0 non-small cell lung cancer. AB - BACKGROUND: Previous studies of patients with surgically resected non-small cell lung cancer and chest wall invasion have shown conflicting results with respect to prognosis. Whether high-risk subsets of the T3 N0 M0 population exist with respect to patterns of failure and overall survival has been difficult to ascertain, owing to small numbers of patients in most series. METHODS AND MATERIALS: A retrospective review was performed to determine patterns of failure and overall survival for patients with completely resected T3 N0 M0 non-small cell lung cancer. From 1979 to 1993, 92 evaluable patients underwent complete resection for T3 N0 M0 non-small cell lung cancer. The following potential prognostic factors were recorded from the history: tumor size, location, grade, histology, patient age, use of adjuvant radiation therapy (18 of 92 patients), and type of surgical procedure (chest wall or extrapleural resection). RESULTS: The actuarial 2- and 4-year overall survival rates for the entire cohort were 48% and 35%, respectively. The actuarial local control at 4 years was 94%. Neither the type of surgical procedure performed nor the addition of thoracic radiation therapy impacted local control or overall survival. CONCLUSION: Patients with completely resected T3 N0 M0 non-small cell lung cancer have similar local control and overall survival irrespective of primary location, type of surgery performed, or use of adjuvant radiation therapy. Additionally, the tumor recurrence rate and overall survival found in this study support the placement of this group of patients in Stage IIB of the 1997 AJCC lung staging classification. PMID- 10477012 TI - Clinical and dosimetric predictors of radiation-induced esophageal toxicity. AB - PURPOSE: To evaluate the incidence, severity, and clinical/dosimetric predictors of acute and chronic esophageal toxicities in patients with non-small cell lung cancer (NSCLC) treated with high-dose conformal thoracic radiation. METHODS AND MATERIALS: Ninety-one patients with localized NSCLC treated definitively with high-dose conformal radiation therapy (RT) at Duke University Medical Center (DUMC) were reviewed. Patient characteristics were as follows: 53 males and 38 females; median age 64 yr (range 46-82); stage I--16, II--3, IIIa--40, IIIb--30, X--2; dysphagia pre-RT--6 (7%). Treatment parameters included: median corrected dose-78.8 Gy (range 64.2-85.6); BID fractionation-58 (64%); chemotherapy-43 (47%). Acute and late esophageal toxicities were graded by RTOG criteria. Using 3D treatment planning tools, the esophagus was contoured in a uniform fashion, the 3D dose distribution calculated (with lung density correction), and the dose volume (DVH) and dose-surface histograms (DSH) generated. At each axial level, the percentage of the esophageal circumference at each dose level was calculated. The length of circumferential esophagus and the maximum circumference treated to doses >50 Gy were assessed. Patient and treatment factors were correlated with acute and chronic esophageal dysfunction using univariate and multivariate logistic regression analyses. RESULTS: There were no acute or late grade 4 or 5 esophageal toxicities. Ten of 91 patients (11%) developed grade 3 acute toxicity. On univariate analysis of clinical parameters, both dysphagia pre-RT (p = 0.10) and BID fractionation (p = 0.11) tended toward significantly predicting grade 3 acute esophagitis. None of the dosimetric parameters analyzed significantly predicted for grade 3 acute esophagitis. Twelve of 66 assessable patients (18%) developed late esophageal toxicity. Of the clinical parameters analyzed, only dysphagia pre-RT (p = 0.06) tended toward significantly predicting late esophageal toxicity. On univariate analyses, the effects of percent organ volume treated >50 Gy (p = 0.05), percent surface area treated >50 Gy (p = 0.05), length of 100% circumference treated >50 Gy (p = 0.04), and maximum percent of circumference treated >80 Gy (p = 0.01) significantly predicted for late toxicity of all grades. On multivariate analysis, percent organ volume treated >50 Gy (p = 0.02) and maximum percent of circumference treated >80 Gy (p = 0.02) predicted for late toxicity. CONCLUSIONS: Late esophageal toxicity following aggressive, high-dose conformal radiotherapy is common but rarely severe. Dosimetric variables addressing the longitudinal and circumferential character of the esophagus have biologic rationale and are predictive of late toxicity. Further studies are needed to assess whether these parameters are better predictors than those derived from traditional DVHs. PMID- 10477013 TI - High dose rate transperineal interstitial brachytherapy for cervical cancer: high pelvic control and low complication rates. AB - PURPOSE: To report the clinical outcome for cervical carcinoma treated with external beam pelvic radiotherapy and interstitial high dose rate (IS-HDR) brachytherapy. METHODS AND MATERIALS: Between July 1991 and June 1996, 62 patients with locally advanced stage cervical carcinoma or early stage carcinoma that precluded satisfactory tandem and ovoid insertion were treated. Most patients received 36 Gy (range: 25 Gy-45 Gy) external beam radiotherapy (EBRT) to the pelvis prior to brachytherapy implant. EBRT was continued, with central shielding, to a dose of 50 Gy to the pelvic sidewalls. HDR Iridium-192 brachytherapy was given in 6 fractions of 5.5 to 6.0 Gy. The mean follow-up was 40 months. RESULTS: Stage distribution was: Stage IB (12), Stage IIA (1), Stage IIB (26), Stage IIIA (6), Stage IIIB (13), and Stage IVA (4). The overall local tumor control was 94%. Local control rates by FIGO stage were Stage I (12/12) 100%, Stage II (25/27) 93%, Stage III (18/19) 95%, and Stage IV (3/4) 75%. The regional pelvic control rates were overall 81%, Stage I (12/12) 100%, Stage II (22/27) 81%, Stage III (15/19) 79%, and Stage IV (1/4) 25%. Distant metastasis developed in 20 patients (32%). The actuarial 5-year disease-free survival was for all patients 48%, Stage I 81%, Stage II 47%, Stage III 39%, and Stage IV O%. Grade 3-4 delayed morbidity resulting from treatment, occurred in 6.5% (4/62) of patients. A fistula without local recurrence occurred in 1.6% (1/62) patients. CONCLUSIONS: We report excellent local and regional pelvic control results using a 6 fraction IS-HDR brachytherapy protocol for cervical carcinoma. The incidence of severe complications is low and suggests that a consistent brachytherapy technique and multiple HDR fractions are therapeutically advantageous to patients treated for cervical carcinoma. PMID- 10477014 TI - Mature results of a pilot study of pelvic radiotherapy with concurrent continuous infusion intra-arterial 5-FU for stage IIIB-IVA squamous cell carcinoma of the cervix. AB - PURPOSE: To evaluate the long-term results of continuous infusion intra-arterial 5-fluorouracil (CI IA 5-FU) given with concurrent pelvic radiotherapy (RT) for FIGO stage IIIB-IVA carcinoma of the cervix. METHODS AND MATERIALS: Between 1965 and 1974, 27 patients with extensive FIGO Stage IIIB (22 patients) or Stage IVA (5 patients) squamous cell carcinoma of the cervix were treated with CI IA 5-FU and RT. Twenty-one patients (78%) had bilateral pelvic wall involvement, 25 (93%) had massive tumors (> or =8 cm in diameter), 7 (27%) had involvement of the lower one-third of the vagina, and 15 (56%) presented with hydronephrosis. All patients underwent routine clinical staging, transperitoneal para-aortic lymph node dissection, and bilateral hypogastric artery catheter placement. 5-FU was continuously infused at a dose rate of 10 mg/kg/day on Days 1-15 of RT. The median dose of 5-FU was 376 mg/m2/day (range 270-692). All patients received concurrent pelvic RT to a median dose of 50 Gy at 2.0 Gy per fraction. Only 4 patients received intracavitary RT. The median follow-up of surviving patients was 190 months. RESULTS: The overall 5-year survival rate was 37%. For the 22 patients with FIGO Stage IIIB disease, the 5-year survival rate was 41%. The survival rate for 18 patients treated with only external beam radiation and chemotherapy for Stage IIIB disease was 33%. Four of 10 patients treated with only 50 Gy of external beam radiation and CI IA 5-FU were long-term survivors. Acute complications, including hematologic toxicity and skin reactions, were severe, with 1 death from neutropenic sepsis. Severe late complications were only observed in patients treated with > or =60 Gy of external beam radiation. CONCLUSIONS: While this series is small, the fact that 4 patients with massive Stage IIIB tumors survived after a total radiation dose of only 50 Gy suggests that RT with CI IA 5-FU deserves further study. Modifications in dose, technique, and route of administration should reduce toxicity, and the addition of intracavitary RT should improve the local effectiveness of combined treatment. PMID- 10477015 TI - Changes in oxygenation during radiotherapy in carcinoma of the cervix. AB - PURPOSE: The aim of this study was to investigate changes in tumor oxygenation, assessed by polarographic needle electrode measurements, following fractionated external beam radiotherapy in carcinoma of the cervix. METHODS AND MATERIALS: Normal and tumor tissue oxygenation was measured in 19 patients prior to radiotherapy and after 40-45 Gy of external beam radiotherapy delivered in 20 fractions over 4 weeks. All measurements were performed during anesthesia. RESULTS: There was no significant difference in the level of normal tissue oxygenation pre- and post radiotherapy. The individual patient median tumor pO2 values ranged from 0 to 31 mmHg pre-radiotherapy and 1 to 61 mmHg post radiotherapy. The mean of the 19 median pO2 values increased from 8 (SD +/- 10) mmHg to 20 (+/- 20) mmHg following external beam radiotherapy. The increase was significant by paired Wilcoxon test (p = 0.011). There was also a significant fall in the proportion of values < 5 mmHg (p = 0.040). Although this value remained constant, or fell, in the majority of patients (15/19), it increased in 4 tumors. Tumor size pre- and postradiotherapy did not correlate with the level of pretreatment oxygenation; neither did the change in tumor size and change in level of oxygenation. CONCLUSION: The level of tumor oxygenation increased in the majority of patients (15/19) following 40-45 Gy of radiotherapy in carcinoma of the cervix. PMID- 10477017 TI - Long-term intellectual outcome in children with posterior fossa tumors according to radiation doses and volumes. AB - PURPOSE: To analyze the relationship between craniospinal irradiation (CSI) and intellectual outcome in children with posterior fossa (PF) tumors. METHODS AND MATERIALS: A neuropsychological evaluation was performed retrospectively in 31 children, aged 5-15 years, who had received radiotherapy for PF tumors, and who had been off therapy for at least 1 year. Factors evaluated for impact on intellectual outcome were: socioeconomic status, disease presentation, histology, complications, chemotherapy, age at radiotherapy, interval between radiotherapy and testing, and radiation doses and volumes. Patients were divided into 3 subgroups according to the CSI doses (0 Gy [i.e., PF irradiation only], 25 Gy, and 35 Gy), with 11, 11, and 9 patients, respectively. RESULTS: Long-term cognitive impairment occurred in most of the patients, even after PF irradiation only. Moreover, there was a significant correlation between the full-scale IQ score (FSIQ) and the CSI dose, with mean FSIQ scores at 84.5 (SD = 14.0), 76.9 (SD = 16.6), and 63.7 (SD = 15.4) for 0 Gy, 25 Gy, and 35 Gy of CS1, respectively. A marked drop in verbal comprehension scores was noted in children who had received the higher dose. CONCLUSION: This preliminary study further supports the rationale for de-escalation of CSI doses and volumes in standard risk PF tumors. PMID- 10477016 TI - A phase II trial of high-dose bromodeoxyuridine with accelerated fractionation radiotherapy followed by procarbazine, lomustine, and vincristine for glioblastoma multiforme. AB - PURPOSE: To conduct a Phase II study to evaluate the long-term efficacy and safety of high-dose 5'-bromodeoxyuridine (BrdU) and accelerated radiotherapy followed by procarbazine, lomustine (CCNU), and vincristine (PCV) chemotherapy in patients with glioblastoma multiforme. METHODS AND MATERIALS: Between 1994 and 1996, 88 patients were enrolled to receive 1.9 Gy of radiation three times a day for two 5-day cycles separated by 2 weeks; each 5-day cycle was preceded by a continuous 96-hour infusion of BrdU at a dose of 2.1 g/m2/day. After radiotherapy, patients received PCV chemotherapy. RESULTS: Median survival for all 88 patients was 50 weeks. Seventy (79.5 %) received one or more courses of PCV; their median survival was 57 weeks. Covariates predictive of improved survival were gross total versus subtotal resection or biopsy (p = 0.0048) and radiation dose > or = 56 Gy (p = 0.019). While receiving BrdU, 47 patients (53%) suffered grade 3 or 4 thrombocytopenia or leukopenia; 22 patients (25%) suffered grade 3 or 4 dermatologic toxicity. CONCLUSION: Survival was not extended in patients with glioblastoma or gliosarcoma who received BrdU at the dose and administration schedule used in this study. The BrdU dose used in this study resulted in substantial myelosuppressive and dermatologic toxicity. PMID- 10477018 TI - Interferon gamma in survivors of the Chernobyl power plant accident: new therapeutic option for radiation-induced fibrosis. AB - BACKGROUND: One of the remarkable clinical consequences of the Chernobyl accident was skin involvement, leading to extensive cutaneous fibrosis. Apart from surgery, no established treatment is available. METHODS: A group of survivors, working in or present at the accident site on April 26, 1986, and a few days thereafter, were examined, treated, and followed-up in 6-month intervals from September 1991 to November 1995. Eight individuals were identified as suffering from excessive cutaneous fibrosis. Skin thickness was measured with high frequency (20 MHz) ultrasound in a clinically well-defined target skin lesion, in addition to histologic confirmation of radiation fibrosis. Interferon gamma was scheduled for all patients on a low-dose regimen (3 x 50 microg/week s.c.). In 2 patients, interferon was discontinued after the first injection, due to withdrawal of consent. In 6 patients, interferon was continued for 30 months, with 1 injection weekly for a further 6 months. Treatment was discontinued in November 1994. Four patients in the treated group and 1 of the 2 patients treated only once ("untreated patients") were reexamined 1 year later. RESULTS: In all individuals treated for 36 months, a significant (p < 0.005) reduction of radiation fibrosis could be determined, in contrast to a significant (p < 0.005) increase in the 2 untreated patients. Follow-up 1 year after discontinuation of the interferon treatment demonstrated significant (p < 0.005) recurrence of fibrosis. CONCLUSION: Low-dose interferon appears to be a safe and effective treatment of cutaneous radiation fibrosis following accidental exposure to high doses of ionizing radiation. Long-term supportive therapy may be required. PMID- 10477019 TI - Radiation oncology training in the United States: report from the Radiation Oncology Resident Training Working Group organized by the Society of Chairman of Academic Radiation Oncology Programs (SCAROP). AB - PURPOSE: In response to the major changes occurring in healthcare, medical education, and cancer research, SCAROP addressed issues related to post-graduate education that could enhance existing programs and complement the present system. METHODS AND MATERIALS: SCAROP brought together a Working Group with a broad range of representatives organized in subcommittees to address: training, curriculum, and model building. RESULTS: The Working Group emphasized the importance of training physicians with the necessary clinical, scientific, and analytical skills, and the need to provide expert radiation oncology services to patients throughout the United States. Opportunities currently exist for graduates in academic medicine, although there may be limited time and financial resources available to support academic pursuits. CONCLUSIONS: In the face of diminishing resources for training and education and the increased scope of knowledge required, a number of models for resident training are considered that can provide flexibility to complement the present system. This report is intended to initiate dialogue among the organizations responsible for radiation oncology resident education so that resident training can continually evolve to meet the needs of cancer patients and take advantage of opportunities for progress through innovative cancer care and research. PMID- 10477020 TI - Description of the role of nonphysician practitioners in radiation oncology. AB - PURPOSE: With changes in reimbursement and a decrease in the number of residents, there is a need to explore new ways of achieving high-quality patient care in radiation oncology. One mechanism is the implementation of nonphysician practitioner roles. The purpose of this paper is to describe the roles and responsibilities of clinical nurse specialists (CNSs), nurse practitioners (NPs), and physician assistants (PAs) currently working in the field of radiation oncology in the United States. METHODS AND MATERIALS: A nationwide mailing was sent to elicit responses to an 8-page self-report questionnaire. RESULTS: The final sample of 86 included 45 (52%) CNSs, 31 (36%) NPs, and 10 (12%) PAs. Two thirds worked in private practice settings. Most of the nonphysician practitioners frequently obtained histories (57-90%) and ordered laboratory studies (52-68%). However, NPs and PAs were more likely than CNSs to frequently perform "medical" services such as perform physical exams (42-80% vs. 19-36%), order radiologic studies (50% vs. 17%), and prescribe medication (60-84% vs. 26%). CNSs were more likely to provide "supportive" services such as develop educational materials, participate in quality improvement initiatives, and develop policies and procedures. CONCLUSIONS: Nonphysician practitioners are not substituting for physicians, but rather are working in collaboration with them, performing designated tasks. PMID- 10477021 TI - Oxygen tension measurements of tumors growing in mice. AB - PURPOSE: Clinical studies using the Eppendorf histograph have shown that patients whose tumors have a low pO2 have worse local control after radiotherapy, and have higher metastatic rates. Because preclinical studies of methods of overcoming, or exploiting, hypoxia generally use transplanted tumors in mice, we have compared the oxygenation of mouse tumors with human tumors to determine the appropriateness of the transplanted mouse model for such preclinical studies. METHODS AND MATERIALS: We evaluated the oxygenation status of subcutaneous (s.c.) tissue and of 12 intradermally (i.d.)- and 7 s.c.-growing mouse or human transplanted tumors in mice using the Eppendorf histograph, and compared the values obtained with measurements of human head and neck nodes. RESULTS: The normal tissue pO2 profile of air-breathing mice showed a nearly Gaussian distribution (38.2+/-14.9 mmHg). Breathing 10% O2 or carbogen resulted in dramatic changes in normal tissue oxygenation. Tumors growing intradermally in the back of air-breathing mice were extremely hypoxic and resistant to expected changes in oxygenation (carbogen breathing, size, and use of anesthetics). Tumors growing s.c. in the foot showed higher oxygen profiles with marked changes in oxygenation when exposing the animals to different levels of oxygen. However, the oxygenation of the mouse tumors transplanted in either site was only a fraction of that of the majority of human tumors. CONCLUSION: Experimental mouse tumors are markedly hypoxic, with median values of 10-20% of those of human tumors. Hence, mouse tumors are probably good models for the most hypoxic human tumors that respond poorly to radiotherapy; however, caution has to be exercised in extrapolating data from mouse to man. PMID- 10477022 TI - Radioprotection of salivary glands by S-2-(3-aminopropylamino) ethylphosphorothioic (amifostine) obtained in a rabbit animal model. AB - BACKGROUND: Impairment of salivary gland function following high-dose radioiodine treatment (HDRIT) is a well-recognized side effect of the treatment. Because differentiated thyroid cancer has an excellent prognosis, reduction of long-term side-effects is mandatory. Therefore, the aim of this study was to investigate the radioprotective effect of amifostine in a rabbit animal model. METHODS: Salivary gland scintigraphy was performed in a total of 16 New Zealand White rabbits. Uptake of 99-Tc-pertechnetate was calculated in percentage of injected activity as a quantitative measure of both salivary gland and thyroid function. Reproducibility of salivary gland scintigraphy was evaluated in one rabbit without any intervention. Fifteen rabbits were studied prior to and up to 6 months after high-dose radioiodine treatment applying 2 GBq 131I. Ten animals received 200 mg/kg amifostine prior to high-dose radioiodine therapy, and 5 served as controls. Salivary glands were examined histopathologically. RESULTS: Variation coefficient of parenchymal function was less than 3.8% in salivary glands. Prior to HDRIT, thyroid uptake was 0.417+/-0.373% and 0.421+/-0.241% in control and amifostine-treated rabbits, respectively. Four weeks after HDRIT, complete ablation of the thyroid was achieved in both groups. Prior to HDRIT, uptake of 99mTc-pertechnetate in salivary glands of five control rabbits was not significantly different from ten amifostine-treated rabbits. In control rabbits 6 months after HDRIT, parenchymal function was reduced significantly (p < 0.0001) by 75.3+/-5.3% and 53.6+/-17.4% in parotid and submandibular glands, respectively. In contrast, in amifostine-treated rabbits, parenchymal function was reduced by 10.6+/-3.4% and 6.5+/-4.3% (p > 0.05) in parotid and submandibular glands, respectively. Histopathologically, marked lipomatosis was observed in control animals but was negligible in amifostine-treated animals. CONCLUSION: Parenchymal damage in salivary glands induced by high-dose radioiodine treatment can be significantly reduced by amifostine in this rabbit animal model. This corresponds to data obtained in patients with differentiated thyroid cancer. PMID- 10477023 TI - Investigation of hypersensitivity to fractionated low-dose radiation exposure. AB - PURPOSE: Hypersensitivity to cell killing of exponentially growing cells exposed to X-rays and gamma rays has been reported for doses below about 0.5 Gy. The reported results have been interpreted to suggest that a dose of 0.5 Gy or less is not sufficient to trigger an inducible repair mechanism. The purpose of this study was to examine this suggested hypersensitivity after multiple low doses (0.3 Gy) of gamma rays where a) the effect would be expected to be significantly magnified, and b) the effect might be of clinical relevance. METHODS AND MATERIALS: C3H 10T1/2 mouse embryo cells were grown to confluence in culture vessels. While in plateau phase of growth, cells were exposed to 6 Gy of gamma rays, delivered in either 6 Gy, 3 Gy, 2 Gy, 1 Gy, or 0.3 Gy well-separated fractions. Corresponding experiments were performed with V-79 and C3H 10T1/2 cells in exponential growth. Cells were replated at low density and assayed for clonogenicity. RESULTS: The results of this study were not inconsistent with some hypersensitivity at low doses, in that 20 fractions each of 0.3 Gy produced a slightly lower (though nonsignificant) surviving fraction compared with the same dose given in 2-Gy fractions. However, the results of the 20 x 0.3 Gy exposures also agreed well with the standard linear-quadratic (LQ) model predictions based on high dose per fraction (1-6 Gy) data. In addition, effects of cellular redistribution were seen which were explained quantitatively with an extended version of the LQ model. CONCLUSIONS: These experiments were specifically designed to magnify and probe possible clinical implications of proposed "low dose hypersensitivity" effects, in which significant deviations at low doses from the LQ model have been suggested. In fact, the results at low doses per fraction were consistent with LQ predictions based on higher dose per fraction data. This finding is in agreement with the well-documented utility of the LQ approach in estimating isoeffect doses for alternative fractionation schemes, and for brachytherapy. PMID- 10477024 TI - Abutment region dosimetry for serial tomotherapy. AB - PURPOSE: A commercial intensity modulated radiation therapy system (Corvus, NOMOS Corp.) is presently used in our clinic to generate optimized dose distributions delivered using a proprietary dynamic multileaf collimator (DMLC) (MIMiC) composed of 20 opposed leaf pairs. On our accelerator (Clinac 600C/D, Varian Associates, Inc.) each MIMiC leaf projects to either 1.00 x 0.84 or 1.00 x 1.70 cm2 (depending on the treatment plan and termed 1 cm or 2 cm mode, respectively). The MIMiC is used to deliver serial (axial) tomotherapy treatment plans, in which the beam is delivered to a nearly cylindrical volume as the DMLC is rotated about the patient. For longer targets, the patient is moved (indexed) between treatments a distance corresponding to the projected leaf width. The treatment relies on precise indexing and a method was developed to measure the precision of indexing devices. A treatment planning study of the dosimetric effects of incorrect patient indexing and concluded that a dose heterogeneity of 10% mm(-1) resulted. Because the results may be sensitive to the dose model accuracy, we conducted a measurement-based investigation of the consequences of incorrect indexing using our accelerator. Although the indexing provides an accurate field abutment along the isocenter, due to beam divergence, hot and cold spots will be produced below and above isocenter, respectively, when less than 300 degree arcs were used. A preliminary study recently determined that for a 290 degree rotation in 1 cm mode, 15% cold and 7% hot spots were delivered to 7 cm above and below isocenter, respectively. This study completes the earlier work by investigating the dose heterogeneity as a function of position relative to the axis of rotation, arc length, and leaf width. The influence of random daily patient positioning errors is also investigated. METHODS AND MATERIALS: Treatment plans were generated using 8.0 cm diameter cylindrical target volumes within a homogeneous rectilinear film phantom. The plans included both 1 and 2 cm mode, optimized for 300 degrees, 240 degrees, and 180 degrees gantry rotations. Coronal oriented films were irradiated throughout the target volumes and scanned using a laser film digitizer. The central target irradiated in 1 cm mode was also used to investigate the effects of incorrect couch indexing. RESULTS: The dose error as a function of couch index error was 25% mm(-1), significantly greater than previously reported. The clinically provided indexing system yielded 0.10 mm indexing precision. The intrinsic dose distributions indicated that more heterogeneous dose distributions resulted from the use of smaller gantry angle ranges and larger leaf projections. Using 300 degrees gantry angle and 1 cm mode yielded 7% hot and 15% cold spots 7 cm below and above isocenter, respectively. When a 180 degree gantry angle was used, the values changed to 22% hot and 27% cold spots for the same locations. The heterogeneities for the 2 cm mode were 70% greater than the corresponding 1 cm values. CONCLUSIONS: While serial tomotherapy is used to deliver highly conformal dose distributions, significant dosimetric factors must be considered before treatment. The patient must be immobilized during treatment to avoid dose heterogeneities caused by incorrect indexing due to patient movement. Even under ideal conditions, beam divergence can cause significant abutment-region dose heterogeneities. The use of larger gantry angle ranges, smaller leaf widths, and appropriate locations of the gantry rotation axis can minimize these effects. PMID- 10477025 TI - Initial clinical experience with a video-based patient positioning system. AB - PURPOSE: To report initial clinical experience with an interactive, video-based patient positioning system that is inexpensive, quick, accurate, and easy to use. METHODS AND MATERIALS: System hardware includes two black-and-white CCD cameras, zoom lenses, and a PC equipped with a frame grabber. Custom software is used to acquire and archive video images, as well as to display real-time subtraction images revealing patient misalignment in multiple views. Two studies are described. In the first study, video is used to document the daily setup histories of 5 head and neck patients. Time-lapse cine loops are generated for each patient and used to diagnose and correct common setup errors. In the second study, 6 twice-daily (BID) head and neck patients are positioned according to the following protocol: at AM setups conventional treatment room lasers are used; at PM setups lasers are used initially and then video is used for 1-2 minutes to fine-tune the patient position. Lateral video images and lateral verification films are registered off-line to compare the distribution of setup errors per patient, with and without video assistance. RESULTS: In the first study, video images were used to determine the accuracy of our conventional head and neck setup technique, i.e., alignment of lightcast marks and surface anatomy to treatment room lasers and the light field. For this initial cohort of patients, errors ranged from sigma = 5 to 7 mm and were patient-specific. Time-lapse cine loops of the images revealed sources of the error, and as a result, our localization techniques and immobilization device were modified to improve setup accuracy. After the improvements, conventional setup errors were reduced to sigma = 3 to 5 mm. In the second study, when a stereo pair of live subtraction images were introduced to perform daily "on-line" setup correction, errors were reduced to sigma = 1 to 3 mm. Results depended on patient health and cooperation and the length of time spent fine-tuning the position. CONCLUSION: An interactive, video based patient positioning system was shown to reduce setup errors to within 1 to 3 mm in head and neck patients, without a significant increase in overall treatment time or labor-intensive procedures. Unlike retrospective portal image analysis, use of two live-video images provides the therapists with immediate feedback and allows for true 3-D positioning and correction of out-of-plane rotation before radiation is delivered. With significant improvement in head and neck alignment and the elimination of setup errors greater than 3 to 5 mm, margins associated with treatment volumes potentially can be reduced, thereby decreasing normal tissue irradiation. PMID- 10477026 TI - Computer-assisted decision making in portal verification--optimization of the neural network approach. AB - PURPOSE: Conventional portal verification requires that a qualified radiation oncologist make decisions as to the set-up acceptability. This scheme is no longer sustainable with the large numbers of images available on-line and stringent time constraints. Therefore the objective of this study was to develop, optimize, and evaluate on clinical data an artificial intelligence decision making tool for portal verification. The tool, based on the artificial neural network (ANN) approach, should approximate, as closely as possible, portal verification assessments made by a radiation oncologist expert. METHODS AND MATERIALS: A total of 328 electronic portal images of tangential breast irradiations were included in the study. A radiation oncologist expert evaluated these images and rated the treatment set-up acceptability on a scale from 0 to 10. Translational and rotational errors in the placement of the radiation field boundaries formed seven-dimensional feature vectors that represented each of the 328 portal images/treatments. The feature vectors were used as inputs to a three layer, feedforward ANN. The neural network was trained on the oncologist's ratings. RESULTS: The rms discrepancy between the ANN and the expert's ratings was 1.05 rating points. Using the decision threshold equal to 5 for both sets of ratings, the ANN classifier was capable of detecting 100% of the portals classified as "unacceptable" by the expert. Only 6.5% of portals acceptable to the oncologist were misclassified as "unacceptable" by the ANN. CONCLUSION: The results of this study indicate the feasibility of using the ANN portal image classifier as an automated assistant to the radiation oncologist. Its role would be to recommend an appropriate decision as to the acceptability or otherwise of a given treatment set-up depicted in a portal image. PMID- 10477027 TI - Portal imaging based definition of the planning target volume during pelvic irradiation for gynecological malignancies. AB - PURPOSE: Geometrical accuracy in patient positioning can vary substantially during external radiotherapy. This study estimated the set-up accuracy during pelvic irradiation for gynecological malignancies for determination of safety margins (planning target volume, PTV). METHODS AND MATERIALS: Based on electronic portal imaging devices (EPID), 25 patients undergoing 4-field pelvic irradiation for gynecological malignancies were analyzed with regard to set-up accuracy during the treatment course. Regularly performed EPID images were used in order to systematically assess the systematic and random component of set-up displacements. Anatomical matching of verification and simulation images was followed by measuring corresponding distances between the central axis and anatomical features. Data analysis of set-up errors referred to the x-, y-,and z axes. Additionally, cumulative frequencies were evaluated. RESULTS: A total of 50 simulation films and 313 verification images were analyzed. For the anterior posterior (AP) beam direction mean deviations along the x- and z-axes were 1.5 mm and -1.9 mm, respectively. Moreover, random errors of 4.8 mm (x-axis) and 3.0 mm (z-axis) were determined. Concerning the latero-lateral treatment fields, the systematic errors along the two axes were calculated to 2.9 mm (y-axis) and -2.0 mm (z-axis) and random errors of 3.8 mm and 3.5 mm were found, respectively. The cumulative frequency of misalignments < or =5 mm showed values of 75% (AP fields) and 72% (latero-lateral fields). With regard to cumulative frequencies < or =10 mm quantification revealed values of 97% for both beam directions. CONCLUSION: During external pelvic irradiation therapy for gynecological malignancies, EPID images on a regular basis revealed acceptable set-up inaccuracies. Safety margins (PTV) of 1 cm appear to be sufficient, accounting for more than 95% of all deviations. PMID- 10477028 TI - A new tool for dose conformity evaluation of radiosurgery treatment plans. AB - PURPOSE: A novel method for dose conformity evaluation of treatment plans produced by the stereotactic radiosurgery treatment planning system is postulated. METHODS AND MATERIALS: By consolidating the information contained in the integral dose-volume histogram and the treatment volume ratio, a plot of treatment volume ratio versus percentage dose may be considered as a useful tool for plan evaluation. To validate the suggested argument, two simple experiments simulating the conformal and nonconformal cases were conducted on the geometric phantom that is commercially available from Radionics. An actual patient treatment plan is also included to explore the effectiveness of the proposed parameters. It is an attempt to establish the baseline of a conformal plan. RESULTS: A plot showed the ability to give the user an idea whether the size of the collimator was adequate to cover the delineated lesion when the user-defined criteria had been in place. Two parameters, namely take-off dose (TOD) and take off volume (TOV) were defined. The former was defined as the maximum dose level found on the surface of the target volume. The TOD is also the maximum possible dose to be received by the adjacent normal tissue. The latter was defined as the percentage of the target volume that received the TOD. Another parameter, irradiated percentage volume (IPV), was defined here as the percentage of the target volume receiving at least the prescribed dose. When the prescribed dose is also the TOD, the IPV becomes the TOV. They were proved to be effective in evaluating the dose conformity. Another term known as equivalent fall-off distance (EFOD) was defined as the equivalent radial distance calculated between two isodose lines. In fact, the dose fall-off rate can also act as a measuring index for plan comparison, because a fast dose fall-off rate is often a requirement for radiosurgery in order to minimize the risk of radiation damage to the surrounding structures. The two phantom studies showed consistent results with the theoretical predictions. The ability of the plot was further explored in the patient treatment plan studies. It was demonstrated that the plot had a remarkable ability to check whether the hot spot is in the vicinity of the lesion. A baseline of a conformal plan was also established; for example, a plan is said to be conformal if its IPV has attained a value of not less than 95% and its associated TVR is not greater than 2. CONCLUSION: The proposed method has demonstrated the effectiveness in dose conformity evaluation. It supplements the integral dose-volume histogram to provide a complete information of a treatment plan in terms of dose uniformity and conformity. PMID- 10477029 TI - A method for implementing the American Association of Physicists in Medicine Task Group-43 dosimetry recommendations for 125I transperineal prostate seed implants on commercial treatment planning systems: regarding Luse et al. IJROBP 37:737 741; 1997. PMID- 10477030 TI - Quantification of the steepness of the dose response relation. PMID- 10477031 TI - Link between local recurrence and distant metastases in human breast cancer. PMID- 10477032 TI - In response to Brock, IJROBP 43:473-474, 1999. PMID- 10477033 TI - Regarding Weiss et al., IJROBP 41:103-109; 1998. PMID- 10477034 TI - What can the investigation of phosphoinositide signaling system in platelets of schizophrenic patients tell us? AB - Disturbances in the regulation of the phosphoinositide signaling system have been proposed as a possible biological marker of schizophrenia. This review considers the laboratory investigations of phosphoinositide metabolism in platelets of schizophrenic patients. We suggest that alterations in the inositol phosphate level and a disturbance of calcium homeostasis may be common denominators for the multiple factors implicated in the pathogenesis of schizophrenia. In addition, these abnormalities may account for the diverse clinical and biochemical manifestations of schizophrenia. PMID- 10477035 TI - Plasma thromboxane and prostacyclin are linear related and increased in patients presenting with acute myocardial infarction. AB - The role of prostanoids in patients with ischemic heart disease including acute myocardial infarction (AMI) has been recognized. However, there is very limited knowledge of the baseline TXB2 and 6-keto-PGF1a plasma levels in patients with AMI before therapy has been administered. We compared plasma levels of TXB2 and 6 keto-PGF1a by enzyme immunoassay in 18 AMI patients before thrombolysis, with those of 13 healthy controls. Plasma levels of TXB2 (319.78+/-16.50 pg/ml) and 6 keto-PGF1a (536.72+/-56.71 pg/ml) were heterogeneous, but significantly higher in the AMI patients than in controls (175.92+/-17.29 pg/ml and 192.08+/-26.11 pg/ml, respectively). In some patients, long-term aspirin therapy mildly inhibits baseline prostanoid levels, however, limited data prevents us from further speculations on this issue. Although, the contributions by prostanoids to the pathogenesis of AMI have been well proposed, their plasma concentrations are not uniformly elevated, and it is still unclear whether the resultant changes are indicative of clinically meaningful effects. PMID- 10477036 TI - The beneficial effects of omega-3 and omega-6 essential fatty acid supplementation on red blood cell rheology. AB - Twenty healthy, non-smoking subjects were enrolled into a study to investigate the effects of dietary supplementation with essential fatty acid (EFAs) on red blood cell rheology. Ten subjects were given 3 months dietary supplementation with long chain polyunsaturated EFAs containing omega-3 and omega-6 EFAs while 10 others were given placebo (sunflower oil). Venous sampling was performed at 0 and 12 weeks and red blood cell (RBC) aggregation and deformability measured by a filtration system. The results showed a reduction in RBC aggregation in the group given omega-3 and omega-6 EFAs but not in the placebo group. This may be related to changes in the RBC membrane and surface receptor characteristics. Such EFAs may be useful in Raynaud's phenomenon. PMID- 10477037 TI - Dose-dependent effects of dietary gamma-linolenic acid on rat spleen lymphocyte functions. AB - Feeding rodents a diet rich in evening primrose oil (EPO), which contains 5-10 g gamma-linolenic acid (GLA)/100 g total fatty acids, has been shown to decrease lymphocyte proliferation and natural killer cell activity. However, EPO contains a very high level of linoleic acid which itself can affect lymphocyte functions and it is not clear to what extent the effects of EPO can be attributed to GLA. The current study investigated the effect of two levels of GLA in the rat diet upon immune cell functions; the level of linoleic acid was maintained below 30 g/100 g total fatty acids. Weanling rats were fed on high fat (178 g/kg) diets which contained 4.4 g or 10 g GLA/100 g total fatty acids in place of a proportion of linoleic acid. The total polyunsaturated fatty acid content and the n-6 to n-3 polyunsaturated fatty acid ratio of the diet were maintained at 35 g/100 g total fatty acids and 7, respectively. The fatty acid compositions of the serum and of spleen leukocytes were markedly influenced by that of the diet, with an increase in the proportions of GLA and dihomo-gamma-linolenic acid when the diets containing GLA were fed; these diets also increased the proportion of arachidonic acid in spleen leukocytes. Spleen lymphocyte proliferation in response to concanavalin A was significantly reduced (by 60%) by feeding the diet containing the higher level of GLA, but not by the diet containing the lower level of GLA. Spleen natural killer cell activity and prostaglandin E (PGE) production by spleen leukocytes were not significantly affected by inclusion of GLA in the diet, although there was a tendency towards decreased natural killer cell activity by cells from rats fed the high GLA diet. Thus, this study shows that dietary GLA is capable of altering the fatty acid composition of cells of the immune system and of exerting some immunomodulatory effects, but that the level of GLA in the diet must exceed 4.4 g/100 g total fatty acids for these effects to become apparent. PMID- 10477038 TI - Polyunsaturated fatty acids inhibit Mg2+ -ATPase in basolateral membranes from rat enterocytes. AB - It is known that certain polyunsaturated fatty acids of the n-6 family, for example linoleic and arachidonic acids, can activate both Na+, K+-ATPase and Ca2+ ATPase. These enzymes drive active absorption processes in the duodenal enterocyte. This study presents data which show a 30-50% inhibition of Mg2+ ATPase activity in enterocyte basolateral membrane preparations by linoleic and gamma-linolenic acids (also a member of the n-6 family.) Mg2+-ATPase activity has several possible roles in the enterocyte: involvement in Mg2+ and Ca2+ absorption (as part of Ca2+-ATPase and also myosin I activity) as well as control of phospholipid distribution in the membrane by a class of Mg2+-ATPases called 'flippases'. The action of linoleic and gamma-linolenic acids on basolateral membrane Mg2+-ATPase may thus modulate several cellular transport processes. PMID- 10477039 TI - Prostaglandin E2 9-keto reductase from bovine term placenta. AB - The aim of the following study was to determine the activity and physico-chemical properties of prostaglandin E2 9-keto reductase from bovine placenta. Placental tissues obtained immediately after parturition were subjected to purification procedure consisting of homogenization, affinity chromatography, gel filtration and allowed to electrophoresis. The activity of enzyme was measured spectrophotometrically. The purification procedures receive 135-fold purified enzyme preparate of the molecular weight of 45 kDa with the following kinetic values: Michaelis constant for PGE2, 117 microM and max velocity 183 pmol/min. The activity of enzyme was also detected with 20 alpha-hydroxypregn-4en-3-one and with 9,10-phenanthrenquinone (Michaelis constant 22 microM and 6 microM, respectively). The determination of physico-chemical properties of prostaglandin E2 9-keto reductase, performed for the first time in bovine placenta, should aid the understanding of the metabolism of prostaglandins and their biological importance in physiological and pathological conditions in cattle. PMID- 10477040 TI - Pretreatment with eicosapentaenoic acid prevented hypoxia/reoxygenation-induced abnormality in endothelial gap junctional intercellular communication through inhibiting the tyrosine kinase activity. AB - Eicosapentaenoic acid (EPA) may protect against atherosclerotic disease, and modulation of endothelium function is one possible mechanism. Hypoxia/reoxygenation (H/R) is a potential risk factor for the pathogenesis of atherosclerosis, and it causes endothelial dysfunction. To evaluate whether EPA may improve the endothelial dysfunction under the condition of H/R, we examined endothelial gap junctional intercellular communication (GJIC), which is said to be important for the endothelium to maintain its normal function. The results indicate that H/R induced a temporal reduction in GJIC after 2 h of reoxygenation in cultured human umbilical vein endothelial cells (HUVEC). This reduction in GJIC was not observed in cells pretreated with 3 microg/ml EPA for 2 days. The results of immunofluorescence show that 2 h reoxygenation caused an increased production of tyrosine-phosphorylated proteins, which was inhibited by EPA pretreatment. Immunoprecipitation demonstrated that tyrosine residues of connexin 43 (Cx43), an important gap junctional protein in HUVEC, were phosphorylated by H/R. However, pretreatment with EPA significantly suppressed this increased phosphorylation. The protective effect of EPA on the reduction in GJIC was also observed in cells treated with 1.5 mM vanadate, a tyrosine phosphatase inhibitor. These data suggest that EPA may ameliorate the H/R-induced GJIC abnormality via inhibition of the tyrosine kinase activation. PMID- 10477041 TI - Nitric oxide in cyclosporine A-induced hypertension: endothelin receptors gene expression. AB - Cyclosporine A (CsA) is an immunosuppressive agent, which also causes hypertension. The effect of CsA on vascular responses was determined in spontaneously hypertensive rats and isolated rat aortic rings. Male rats weighing 250-300 g were given either CsA (25 mg/kg/day) in olive oil or vehicle by i.p. injection for 7 days. CsA administration produced a 27% increase (P < 0.001) in mean arterial pressure (MAP) which reached a plateau after 3 days. Conversely, the level of nitrate/nitrite, metabolites of nitric oxide (NO), decreased by 44% (P < 0.001) in the urine. In the presence of endothelin (ET) 10(-9) M, thoracic aortic rings from rats treated with olive oil, L-Arginine (L-Arg) or L-Arg+CsA showed a 100% increase (P < 0.001) in tension compared to the aortic rings from rats treated with CsA alone; aortic rings from rats treated with CsA alone did not respond to ET. The effects of CsA were reversed in both in vivo and in vitro by pretreatment with L-Arg (10 mg/kg/day ip), the precursor of NO. There were no changes in MAP and tension in rats treated with L-Arg alone. Possible explanation for lack of response to ET of aortic rings from CsA treated rats may be that CsA affected ET signalling pathway; ET receptors mRNA (messenger ribonucleic acid) gene expression was inhibited in aortic rings of rats treated with CsA. In summary, CsA inhibits endothelial NO formation, with resulting increases in MAP, and this inhibition can be overcome by parenteral administration of L-Arg. PMID- 10477042 TI - Endothelin-induced prostacyclin production in rat aortic endothelial cells: role of calcium. AB - Endothelin (ET) is a potent vasoconstrictor peptide, released from endothelial cells, which is associated with prostaglandin (PG) release. The mechanism by which ET causes the release of PG is not clearly understood. We used rat aortic endothelial cells to investigate the role of calcium (Ca2+) in ET-1-induced prostacyclin (PGI2) release. ET-1 (10(-9) M) produced a significant increase in PGI2 release. Pretreatment of rat aortic endothelial cells with different doses (10(-9) M and 10(-6) M) of diltiazem (voltage-sensitive L-type calcium channel blocker) produced significant inhibition of ET-1- and PDBu-induced PGI2 release. Inhibition was first noted at 10(-9) M and was complete at 10(-6) M. Conversely, pretreatment of rat aortic endothelial cells with different doses (10(-9) M and 10(-6) M) of calcium channel blockers (thapsigargin, an intracellular calcium channel blocker or conotoxin, a voltage-sensitive N-type calcium channel blocker) produced no changes on ET-1- or PDBu-induced PGI2 release. These results provide further support for the concept that PKC mediates ET-induced PGI2 release in rat aortic endothelial cells via an increase in intracellular calcium and this increase is due to the influx of extracellular calcium and not to the release of calcium from the sarcoplasmic reticulum. PMID- 10477043 TI - Eicosapentaenoic acid modulates arachidonic acid metabolism in rat alveolar macrophages activated by silica. AB - Eicosapentaenoic acid (EPA) was used to modulate the activation of alveolar macrophages, to examine its potential anti-inflammatory effect in addition to its anti-arteriosclerotic or anti-thrombotic effects. Wistar strain rat alveolar macrophages (2 x 10(6) cell) obtained by bronchoalveolar lavage were preincubated with EPA (0-20 microM), and further incubated with 1 mg of silica for 90 min. Leukotriene (LT) B4 and LTB5 of the supernatant were analyzed by reverse phase HPLC. EPA inhibited the production of LTB4 dose-dependently. The production of LTB5, a metabolite from EPA, was increased at low concentrations of EPA (0-10 microM) and decreased at high concentrations (>10 microM). These results suggest that EPA is competitive with arachidonic acid (AA) at low concentrations, and that EPA may inhibit AA metabolism via inhibition of 5-lipoxygenase or phospholipase A2 at high concentrations. PMID- 10477044 TI - In vitro and in vivo pharmacological characterization of SB 201993, an eicosanoid like LTB4 receptor antagonist with anti-inflammatory activity. AB - Leukotriene B4 (LTB4) and 12-(R)-hydroxy-5,8,10,14-eicosatetraenoic acid (12-[R] HETE) have been postulated to contribute to the pathophysiology of inflammatory diseases. SB 201993, (E)-3-[[[[6-(2-carboxyethenyl)-5-[[8-(4-methoxyphenyl)octyl] oxy]-2-pyridinyl] methyl] thio] methyl] benzoic acid, identified from a chemical series designed as ring-fused analogs of LTB4, was evaluated as an antagonist of LTB4- and 12-(R)-HETE-induced responses in vitro and for anti-inflammatory activity in vivo. SB 201993 competitively antagonized [3-H]-LTB4 binding to intact human neutrophils (Ki = 7.6 nM) and to membranes of RBL 2H3 cells expressing the LTB4 receptor (RBL 2H3-LTB4R; IC50 = 154 nM). This compound demonstrated competitive antagonism of LTB4- and 12-(R)-HETE-induced Ca2+ mobilization responses in human neutrophils (IC50s of 131 nM and 105 nM, respectively) and inhibited LTB4-induced Ca2+ mobilization in human cultured keratinocytes (IC50 = 61 nM), RBL 2H3-LTB4R cells (IC50 = 255 nM) and mouse neutrophils (IC50 = 410 nM). SB 201993 showed weak LTD4-receptor binding affinity (Ki = 1.9 microM) and inhibited 5-lipoxygenase (IC50 of 3.6 microM), both in vitro and ex vivo. In vivo, SB 201993 inhibited LTB4-induced neutrophil infiltration in mouse skin and produced dose-related, long lasting topical anti inflammatory activity against the fluid and cellular phases of arachidonic acid induced mouse ear inflammation (ED50 of 580 microg/ear and 390 microg/ear, respectively). Similarly, anti-inflammatory activity was also observed in the murine phorbol ester-induced cutaneous inflammation model (ED50 of 770 and 730 microg/ear, respectively, against the fluid and cellular phases). These results indicate that SB 201993 blocks the actions of LTB4 and 12-(R)-HETE and inhibits a variety of inflammatory responses; and thus may be a useful compound to evaluate the role of these mediators in disease models. PMID- 10477045 TI - A nitric oxide synthase inhibitor, L-NAME, attenuates saccharin drinking in a two choice test in water-deprived rats. AB - The nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L NAME) [0 (veh)], 10, 25, and 50 mg/kg s.c.) was administered to water-deprived, saccharin-preferring, rats in a 30-min two-bottle choice test of 0.1% sodium saccharin and tap water in a within subjects design. Saccharin intake was selectively attenuated in a dose-related manner with increasing dose of L-NAME, reaching statistical significance at 25 and 50 mg/kg L-NAME when compared to vehicle control condition (p < 0.01). In contrast, water intake was not appreciably affected. Total fluid intake was attenuated as well. Neither saccharin nor water intake in a second group of animals was significantly affected by the inactive isomer, D-NAME, suggesting a stereospecific action. These data suggest that a taste factor might contribute to the well-documented hypophagic action of NOS inhibitors in a number of animal species. The possibility that such effect might be mediated through a serotonergic mechanism is considered. PMID- 10477046 TI - Effects of restricted food access on diurnal fluctuation of behaviors and biochemical functions in hereditary microphthalmic rats. AB - The characteristics in circadian rhythms of spontaneous locomotor activity, and some metabolic properties were examined in microphthalmic mutant rats of the Donryu strain under ad lib or restricted food access conditions. The growth of microphthalmic rats was retarded compared to that of normal-sighted rats from the same strain. Under a 12:12-h light:dark (LD) cycle with free access to food, normal-sighted rats showed basically nocturnal patterns of the locomotor activity rhythms, but most of microphthalmic rats manifested free-running rhythms and a few of them showed arrhythmic. When food access was restricted only for 6 h in the light period of the LD cycle, the normal and hereditary blind rats generated gradually new patterns of the locomotor activities in which the animals showed to be more active in the light period. Plasma glucose concentration in normal rats showed a peak after food consumption, but microphthalmic mutants exhibited no periodic changes of the glucose levels. Responses of the biochemical parameters of protein and mineral metabolism to restricted food access in the mutants did not differ from those in normal rats. These results suggest that microphthalmic mutant rats show the free-running circadian rhythm of locomotor activity due to a complete lack of their optic nerve and visual input to the circadian clock, but the mutants maintained the ability to shift their circadian phase induced by restricted food access similar to that in control rats, and also that the mutants have almost normal properties of biochemical and physiological functions except for glucose metabolism. PMID- 10477047 TI - Synergistic anorectic effect of dehydroepiandrosterone and d-Fenfluramine on the obese Zucker rat. AB - Fasted obese, female Zucker rats accustomed to eating a single high fat meal within 1 h a day were treated with intraperitoneal injections of dehydroepiandrosterone (DHEA) and dextrofenfluramine (d-fen), either individually or in combination. Caloric intake was measured over a 1-h period 2 h after drug administration, and results compared to that of vehicle-treated controls. At 50 mg/kg body weight, DHEA did not affect food intake. At doses of < or = 2 mg/kg d fen did not affect food intake. Together, however, DHEA 50 mg/kg and d-fen < or = 2 mg/kg significantly decreased food intake. At doses of > or = 3 mg/kg d-fen diminished caloric intake by itself, and the addition of DHEA significantly augmented this effect. Neurotransmitter levels in select regions of the hypothalamus of animals treated using a similar drug protocol showed several changes in the levels of serotonin and its metabolite 5 hydroxyindole acetic acid (5-HIAA). It is hypothesized that DHEA augments the production of serotonin while d-fenfluramine enhances its release, and together these two actions may account for the synergistic action of DHEA and d-fenfluramine. PMID- 10477048 TI - Morning sickness and salt intake, food cravings, and food aversions. AB - Evidence for an association between early pregnancy sickness and offspring salt (NaCl) preference has been obtained from studying offspring as young adults and as infants. To determine whether the association between early pregnancy sickness and salt preference of offspring is secondary to familiar similarity in salt preference, the present study examined the self-reported salt intake and dietary cravings and aversions of pregnant women. Women who reported little or no vomiting (n = 108) were compared to women who reported moderate to severe vomiting (n = 21) during pregnancy. The women's self-reported salt use and reported cravings and aversions for common food were measured via survey for time periods prior to and during their current pregnancy. Women did not differ in reported salt use prior to pregnancy as a function of their pregnancy symptoms. Women reported more aversions during, than prior to, pregnancy (p < 0.05). Women with more severe vomiting reported a greater number of aversions (p < 0.05) both prior to and during pregnancy. There was a significant association between experiencing cravings and aversions prior to pregnancy and experiencing craving and aversions during pregnancy (p < 0.05). These findings do not provide evidence for an association between dietary levels of sodium and the likelihood of experiencing severe pregnancy symptoms. Therefore, these data do not support the suggestion that reported elevations in salt preference in offspring of women with moderate to severe vomiting during pregnancy are mediated by familial dietary practices. PMID- 10477049 TI - Effect of predatory stress on sucrose intake and behavior on the plus-maze in male mice. AB - In this study, the effect of the exposure of male mice to sensory stimuli from rats was assessed on both sucrose intake and the elevated plus-maze tests. CDl male mice were trained in the sucrose intake task (the prestress phase) and, subsequently, distributed into two groups. The stressed group was accommodated in the same room as rats and the control group with mice (the stress phase). After being transferred, animals were tested on sucrose intake and the plus-maze (acute tests) and retested three times a week for sucrose intake and once on plus-maze on the last day (chronic tests). After acute exposure to the predator, the only difference between stressed and control animals was a higher number of fecal boli left on the plus-maze by the former. During the chronic phase, stressed animals showed a lower level of sucrose intake and higher level of anxiety than controls. In conclusion, this study shows that chronic exposure of male mice to stimuli from rats reduces the sensitivity to the rewarding properties of sucrose and prevents the habituation to the plus-maze observed in controls. Thus, this study suggests that exposure of mice to sensory stimuli from rats may provide an animal model of stress, and that these species should not be routinely housed together. PMID- 10477050 TI - Previous training in the water maze: differential effects in NMRI and C57BL mice. AB - It has been shown that acquisition rates in the water maze vary across strains of mice, although the differential effects of previous experience in this spatial task have been scarcely evaluated. The aim of the present study was to evaluate the effects of training in the water maze at an early age (2 months) in two strains of mice (NMRI and C57BL) using a longitudinal study. Mice with or without previous training were tested when they were 6 months, and retested when 10 months old. The results showed that trained NMRI mice performed better than all the other groups, both at test and retest, indicating that previous training had more beneficial effects in NMRI than in C57BL mice. These results demonstrate that the effects of an early training in the water maze may be influenced by the characteristics of the strain of mice. It could have implications in longitudinal studies evaluating effects of pharmacological or behavioral manipulations. PMID- 10477051 TI - Effects of aging on the acquisition and extinction of excitatory conditioning in Drosophila melanogaster. AB - Young (7-day-old), middle-aged (28-day-old), and old (49-day-old) Drosophila melanogaster were compared for acquisition, then extinction, of excitatory conditioning of the Proboscis Extension Response. The contribution of nonassociative processes (sucrose-induced Central Excitatory State, Pseudoconditioning) in the elicitation of conditioned responses was simultaneously assessed. Old flies displayed a faster and stronger acquisition than middle-aged ones, whereas no significant acquisition was stated in young flies. The influence of nonassociative processes on the acquisition was of minor importance, even though Central Excitatory State increased with age. Old flies proved also to be the slowest to extinguish. Because nonassociative contamination was not involved during extinction, one may wonder whether extinction was delayed in old flies mainly due to a higher level of acquisition or to a behavioral rigidity. PMID- 10477052 TI - Effects of the 5-HT1A receptor agonist 8-OH-DPAT on operant food intake in food deprived pigs. AB - The effects of the 5-HT1A agonist 8-hydroxy-2 (di-n-propylamino)tetralin (8-OH DPAT) were investigated on operant food intake in food-deprived pigs. In Experiment 1, 8-OH-DPAT (5-20 microg/kg) administered intravenously (i.v.) 15 min prior to the occurrence of feeding produced a dose-related decrease in operant food intake in pigs that had been fasted overnight. The effects were mainly apparent during the first 30 min after the start of the feeding period. In Experiment 2, 8-OH-DPAT (25 and 50 microg/kg, i.v.) administered 60 min prior to the occurrence of feeding in pigs that were fasted overnight also produced significant decreases in food intake. The effects were mainly apparent during the first 30-40 min after the start of the feeding period. In Experiment 3, 8-OH-DPAT (20 microg/kg, i.v.) significantly increased operant feeding in satiated pigs during the first 30 min after administration. These results show that 8-OH-DPAT has complex effects on feeding behaviour in pigs, increasing operant food intake in satiated pigs, while producing a reduction in food intake in food-deprived animals. PMID- 10477053 TI - Negative affect and voluntary alcohol consumption in Wistar-Kyoto (WKY) and Sprague-Dawley rats. AB - Based on the assumption that the Wistar-Kyoto (WKY) rat strain represents an animal model for depressive behavior, the purported relationship between depression and alcohol consumption was investigated in three experiments. WKY rats consumed more alcohol than Sprague-Dawley (S-D) rats when offered a choice between a 7% alcohol solution and tap water. Subsequently, the severity of stress induced stomach ulcers was significantly less in WKY rats that had access to alcohol. In Experiment 2, WKY and S-D rats were assigned to either an alcohol access treatment or to a water-only treatment for 27 days and subsequently observed in the open-field test (OFT) and the elevated plus-maze (EPM). Access to alcohol reduced response latency in the OFT, and increased the percent time in the open arm and the total number of arm entries in the EPM for WKY rats. In Experiment 3, the antidepressant, imipramine, reduced alcohol consumption in both strains and significantly increased percent time in the open arms of the EPM for WKY rats. These studies support the assumption that depression and alcohol consumption may be related. PMID- 10477054 TI - Conditioned flavor avoidance, preference, and indifference produced by intragastric infusions of galactose, glucose, and fructose in rats. AB - The postingestive reinforcing and satiating effects of intragastric (i.g.) infusions of 16% galactose, glucose, and fructose were compared in adult female rats. In Experiment 1, food-restricted rats were trained to drink (30 min/day) flavored solutions (the CS+Gal and CS+Glu) paired with intragastric (i.g.) infusions of galactose and glucose; other flavors (the CS-) were paired with IG water infusions. In subsequent choice tests, the rats strongly preferred (91%) the CS+Glu to the CS-, but avoided the CS+Gal (21%) in favor of the CS-. In Experiment 2, the rats were trained with a CS+Fru paired with i.g. fructose infusions and a CS- paired with i.g. water. In the choice test they consumed similar amounts of CS+Fru and CS- (CS+Fru preference = 51%). In other choice tests they preferred the CS+Glu (>80%) to CS+Fru and CS+Gal, and the CS+Fru (81%) to CS+Gal. Satiation tests were performed in Experiment 3 by adapting the rats to drink a 3% sugar + 0.2% saccharin solution paired with i.g. water infusions (30 min/day). On different test days 16% sugar instead of water was infused. IG galactose, glucose, and fructose produced comparable reductions in sugar+saccharin intake in the first test session. These findings demonstrate that, while the three sugars had similar satiating effects, they differed substantially in their postingestive flavor conditioning effects. The glucose and fructose results confirm prior data indicating that only glucose generates potent postingestive reinforcing stimuli. The galactose-induced flavor avoidance indicates that this sugar has a negative postingestive consequence. This may be due to the slow and incomplete hepatic metabolism of this sugar in adult rats. Conceivably, galactose intolerance may contribute to the lactose avoidance in adult animals. PMID- 10477055 TI - Control of motor seizures by brotizolam with maintenance of stable refractory periods for self-stimulation. AB - In recent years, we have been pursuing our mapping investigations of the substrate for brain-stimulation reward in regions of the anterior hypothalamic and lateral preoptic areas. However, one problem is that stimulation of these sites often generates overt seizures so that their suppression via a pharmacological means would be very useful. The sedative-hypnotic benzodiazepine, brotizolam, is reportedly a long-lasting anticonvulsant. Hence, its effects on motor seizures elicited from stimulation of the lateral preoptic area were evaluated in the first experiment. Both tested doses (5.0 and 7.5 mg/kg) of the drug were shown to significantly decrease the number, and marginally, the severity of stimulation-induced seizures; furthermore, this effect was relatively long lasting, up to about 3 h. The higher dose of brotizolam did not alter the single-pulse thresholds for self-stimulation, a requirement for evaluations of poststimulation excitability, the purpose of the second experiment. Here, our interest was in documenting whether the membrane properties of the stimulated neurons, as assessed by refractory periods, were altered by brotizolam. No differences in the time course of recovery were observed; refractoriness began between 0.4 and 0.8 ms, and reached 50% recovery by 2.0 ms, which is consistent with the pattern of poststimulation excitability typically measured at these sites. Thus, in addition to its long-lasting suppression of motor seizures in rats, brotizolam does not alter the time course of recovery from refractoriness of the neurons that mediate brain-stimulation reward in the lateral preoptic area. PMID- 10477056 TI - Effects of food deprivation and metabolic fuel utilization on food hoarding by jirds (Meriones shawi). AB - Food hoarding plays an important role in the energetic repertoire of a variety of mammalian species. Both food hoarding and food intake have been examined in rodents using several energetic challenges including food deprivation, treatment with metabolic fuel blockers, and enhancement of fuel storage. In the present experiment, we examined food hoarding by female jirds (Meriones shawi), a desert rodent species occupying the arid steppes and desert regions of Egypt. Jirds are prodigious hoarders in the field; however, virtually nothing is known about their hoarding within controlled laboratory settings. In the present study, the effects of food deprivation as well as alterations in metabolic fuel utilization (i.e., 2 deoxy-D-glucose and isophane insulin) on food hoarding and food intake were tested in female jirds using a simulated burrow system. Jirds decreased body mass and increased food consumption following either 32 or 56-h food deprivation. Food hoarding, however, was virtually abolished after food deprivation and treatment with 2-DG. In contrast, isophane insulin treatment had no effect on food consumption or hoarding in this species. Taken together, the present results suggest that total body mass (fat), rather than short-term metabolic fuel utilization, regulates both food consumption and hoarding in female jirds. In addition, these results provide a novel set of appetitive responses to these energetic challenges in small mammals. PMID- 10477057 TI - Heritability of hunger relationships with food intake in free-living humans. AB - The heritability of the before and after meal self-ratings of hunger and thirst and their relationship to food intake was investigated with 110 identical and 102 fraternal same-sex and 53 fraternal mixed-gender adult twin pairs who were paid to maintain 7-day food intake diaries. From the diary reports, the total and meal intakes of food energy and the amounts of the macronutrients ingested were estimated. Subjects rated their subjective levels of hunger on a seven-point scale both before and after eating. Linear structural modeling was applied to investigate the nature and degree of genetic and environmental influences and revealed significant genetic influences on the self-rated levels of hunger both before and after the meals and the change in hunger over the meal. In addition, the relationship between hunger and intake was influenced by the genes. Significant heritabilities were found for the correlations, and the slopes of the regression lines relating before-meal hunger to meal size and also the change in hunger over the meal to intake. This indicates that hunger's effect on intake and, in turn, intake's effects on hunger are to some extent heritable. This suggests that the level of subjective experience and the individuals responses to them are influenced by the genes and become part of the total package of genetically determined physiological, socio/cultural, and psychological processes that regulate energy balance. PMID- 10477058 TI - The effects on feeding of galanin and M40 when injected into the nucleus of the solitary tract, the lateral parabrachial nucleus, and the third ventricle. AB - Several reports indicate that central injection of galanin stimulates feeding, and that there is macronutrient specificity in this response. In addition, the galanin receptor antagonist, M40, reduces food intake when injected centrally. The nucleus of the solitary tract (NTS) and the lateral parabrachial nucleus (PBN) contain galanin receptors, and are involved in the control of food intake. Hence, we sought to compare the feeding response to galanin injection into these areas with that of third ventricle (3V) galanin injection. The feeding response to injection of galanin was greatest for the 3V. Hindbrain injection of galanin stimulated food intake only at the beginning of the dark period. NTS injection of M40 inhibited intake of a macronutrient diet in food-deprived rats, but was ineffective at reducing dark-onset feeding or deprivation-induced chow intake. 3V injection of M40 did not reduce deprivation-induced intake. PBN injection of galanin at dark onset had no effect in a group of fat-preferring rats. These results suggest that hindbrain galanin may contribute to feeding by inhibiting satiety, and that hypothalamic galanin receptors are involved with stimulation of intake. Furthermore, the absence of a consistent pattern of the stimulation of macronutrient intake suggests that galanin may not be a significant effector of macronutrient selection during individual meals. PMID- 10477059 TI - Chemosensory detectability of 1-butanol and 2-heptanone singly and in binary mixtures. AB - Using 1-butanol and 2-heptanone as stimuli, we measured detectability (i.e., psychometric) functions for the odor, nasal pungency, and eye irritation of these two substances alone and in binary mixtures. Nasal pungency responses were tested in subjects lacking olfaction (i.e., anosmics) for whom odors do not interfere. Eye irritation responses were tested in normosmics and anosmics, and found to be similar in both groups so their results were pooled. When all stimuli--single and mixtures--were transformed into concentration units of one (or the other) chemical, a single function could fit all data from the same sensory end point with a correlation coefficient of 0.91 or higher. The outcome lends support, as a first approximation, to the notion of chemosensory agonism, in the sense of dose additivity, between the members of binary mixtures presented at perithreshold levels. PMID- 10477060 TI - Water-deprivation prevents morphine-, but not LiCl-induced, suppression of sucrose intake. AB - Intake of a saccharin-conditioned stimulus (CS) can be suppressed following pairing with an aversive agent such as lithium chloride (LiCl) or x-rays (referred to as a conditioned taste aversion or CTA), a highly rewarding sucrose solution (referred to as an anticipatory contrast effect), or a drug of abuse such as morphine or cocaine. Although the suppressive effects of LiCl and sucrose are clear examples of aversive and appetitive conditioning, respectively, it is not certain which properties (aversive or appetitive) mediate the suppressive effects of drugs of abuse. It is known, however, that the suppressive effects of a rewarding sucrose US are attenuated when using a caloric sucrose CS in food deprived rats, while LiCl induced CTAs are much less effected. Standard CTA testing typically is conducted in water-deprived rather than food-deprived rats and, although LiCl is known to suppress intake of a sucrose CS in water-deprived rats, the suppressive effects of drugs of abuse have not been evaluated under these conditions. The present experiment, then, compared the suppressive effects of a standard dose of morphine (15 mg/kg) and a matched dose of LiCl (0.009 M) on intake of a sucrose CS in water-deprived and free-feeding rats. The results showed that both drugs suppressed intake in free-feeding subjects, but only the aversive agent, LiCl, reduced CS intake in the water-deprived rats. This finding dissociates the suppressive effects of morphine and LiCl and, in so doing, aligns the suppressive effects of morphine with those of an appetitive sucrose US. PMID- 10477061 TI - Chorda tympani responses in two inbred strains of mice with different taste preferences. AB - Behavioral studies suggest that there are significant differences in the taste systems of the inbred mouse (Mus musculus) strains: C57BL/6J (B6) and DBA/2J (D2). In an attempt to understand the biological basis of the behavioral differences, we recorded whole-nerve chorda tympani responses to taste solutions and compared the results to intake of similar solutions in nondeprived mice. Stimuli included a test series composed of 0.1 M sodium chloride, 0.3 M sucrose, 10 mM sodium saccharin, 3 mM hydrochloric acid, and 3 mM quinine hydrochloride, as well as concentration series for the same substances. Neural activity of the chorda tympani that was evoked by sucrose, saccharin, or NaCl was greater in B6 than D2 mice; and neural threshold for sucrose was lower in B6 mice, but neural thresholds for HCl and quinine were lower in D2 mice. B6 mice drank more sucrose and saccharin but less quinine than D2 mice; thus, sucrose and saccharin preference were positively correlated, but NaCl and quinine aversiveness were negatively correlated with the chorda tympani results. Nonetheless, genes involved in the structuring of taste receptors and/or the chordae tympani, which transduce taste stimuli having diverse perceptual qualities, differ for the two mouse strains. PMID- 10477062 TI - Comparison of the effects of a high-fat and high-carbohydrate soup delivered orally and intragastrically on gastric emptying, appetite, and eating behaviour. AB - To investigate the effects of fat and carbohydrate on appetite, food intake and gastric emptying with and without the influence of orosensory factors, a group of nine healthy, fasted male subjects took part in two separate paired experiments involving high-fat and high-carbohydrate radiolabelled soup preloads. In the first experiment subjects received direct intragastric isocaloric infusions of either a high-fat tomato soup or a high-carbohydrate tomato soup (400 kcal in 425 mL) over 15 min, on two occasions. In the second paired experiment subjects ingested the same high-fat and high-carbohydrate soup over 15 min. In both experiments ratings of hunger and fullness were recorded over a period of 135 min and gastric emptying was measured by scintigraphy. Food intake was evaluated from a test meal (yoghurt drink) given 2 h after the end of the soup infusion/ingestion. When soup was administered intragastrically (Experiment 1) both the high-fat and high-carbohydrate soup preloads suppressed appetite ratings from baseline, but there were no differences in ratings of hunger and fullness, food intake from the test meal, or rate of gastric emptying between the two soup preloads. When the same soups were ingested (Experiment 2), the high-fat soup suppressed hunger, induced fullness, and slowed gastric emptying more than the high-carbohydrate soup and also tended to be more effective at reducing energy intake from the test meal. The results of these studies demonstrate that orosensory stimulation plays an important role in appetite regulation, and also indicate that subtle differences in orosensory stimulation produced by particular nutrients may profoundly influence appetite and gastrointestinal responses. PMID- 10477063 TI - Food entrainment to 4-h T cycles in rats kept under constant lighting conditions. AB - The effect of 4-h feeding cycles on the feeding pattern of rats kept under constant light and constant darkness, was analysed. In both cases, the scheduled pattern of food-approach behavior elicited dissociation of the feeding activity into several components associated to the feeding times (CAFT), which coexisted with the previous circadian free-running rhythms. In LL rats and in the rest phase of DD animals, the CAFT was characterized by an increased number of food approaches confined exclusively to the period of food availability, with poor anticipatory activity. However, in the active phase of some DD rats, CAFT included an additional anticipatory activity. The interaction between the CAFT and the free-running rhythms involved modulation of the CAFT, depending on the free-running periodicity. After termination of the feeding-restriction schedule, some DD and LL animals retained the CAFT for at least 3 days. However, when the access to food was blocked 10 days after ending the restricted schedule, rats did not show any feeding activity associated to the previous feeding times. A spontaneous feeding pattern similar to that imposed by the previous feeding schedule emerged immediately after food deprivation in two of the DD animals. Although these results are compatible with the existence of a food-entrainable pacemaker, the existence of a multioscillatory light-entrainable pacemaker with some oscillators entrained by food pulses and others free-running may explain our results. PMID- 10477064 TI - Survival of multidrug-resistant Salmonella typhimurium DT104 in egg powders as affected by water activity and temperature. AB - A study was done to determine if a four-strain mixture of multidrug-resistant Salmonella typhimurium definitive type 104 (DT104) cells and a four-strain mixture of S. typhimurium non-DT104 cells differed in ability to survive in whole egg powder, whole egg powder supplemented with corn syrup solids (38%) and salt (1.9%), egg yolk powder, and egg white powder as affected by a(w)(0.29-0.37 and 0.51-0.61) during storage at 13 or 37 degrees C for 8 weeks. Rates of inactivation of S. typhimurium DT104 and non-DT104 cells were similar within each set of test parameters. With the exception of whole egg powder supplemented with corn syrup solids and salt, death was enhanced at a(w) 0.29-0.37 compared to a(w) 0.51-0.61 when powders were stored at 13 degrees C. Survival of cells in whole egg powder supplemented with corn syrup solids and salt was significantly (P < or = 0.05) higher compared to survival in other egg powders stored at 13 degrees C. The opposite trend occurred in powders at a(w) 0.5-0.61 stored at 37 degrees C. Survival of S. typhimurium DT104 and non-DT104 cells at initial populations of 5.01-5.39 log10 cfu/g of egg white powder containing 4.9, 6.1, or 8.2% moisture at 54 or 82 degrees C for 7 days or 8 h, respectively, was determined. Rates of inactivation of DT104 and non-DT104 cells did not differ. Both cell types were detected in egg white powder containing 4.9% moisture but not in powder containing 8.2% moisture when held at 54 degrees C for 7 days. Heating at 82 degrees C for 8 h failed to eliminate 5 log10 S. typhimurium per g of egg white powder, regardless of the moisture content. PMID- 10477065 TI - Flow cytometry and RT-PCR for rotavirus detection in artificially seeded oyster meat. AB - A flow cytometry (FC)-based method was developed for the detection of rotavirus in oyster meat using simian rotavirus SA11 as a model. To study virus recovery, oyster meat was injected with rotavirus and the oyster extract used to infect MA104 cell monolayers. Following varying periods of infection, the cells were recovered and reacted with the monoclonal antibody M60 which is specific for the rotavirus group A serotypes 1-4 outer capsid protein, VP7, followed by a second antibody (anti mouse IgG-FITC). A FACScan FC was used to estimate the number of infected cells as well as the level of infection. To evaluate the sensitivity of the method, non-inoculated oysters were processed following the same extraction protocol and, at the end, they were seeded with the same amount of virus used for oyster inoculation. This seeded oyster extract was then used to infect MA104 cells and the number of infected cells determined using the same FC procedure. A semi-nested two-step PCR for detection of rotavirus nucleic acid was undertaken to compare the sensitivity of FC with RT-PCR. Using FC, as little as 0.02 flow cytometry units (fcu) (number of infected cells counted by FC) could be detected after 72 h of cell infection. This is a very similar limit of sensitivity to that obtained with RT-PCR. Both methods are approximately 100 times more sensitive than the plaque-forming units (pfu) assay. PMID- 10477066 TI - The effect of waste water reuse in irrigation on the contamination level of food crops by Giardia cysts and Ascaris eggs. AB - In Marrakech, raw sewage has been used for farming purposes for several decades for many types of crops. This study aimed to determine the contamination level of Giardia cysts and Ascaris eggs for crops designated for human consumption. Collected crops in irrigated fields were turnip, marrow, squash, potatoes, pepper and eggplant. Field trials were also carried out on four crops, coriander, carrots, mint and radish, using three water types for irrigation, i.e. raw waste water, treated waste water (sedimentation and 16 days retention) and fresh water. Giardia cysts were detected at a level of 5.1 cysts/kg in potatoes, while Ascaris eggs were observed in numbers varying between 0.18 eggs/kg in potatoes and 0.27 eggs/kg in turnip. Field trials confirmed that irrigation of crops by raw waste water leads to contamination. Giardia and Ascaris were isolated in coriander at concentrations of 254 cysts/kg and 2.7 eggs/kg, respectively; mint was also highly contaminated with numbers reaching 96 cysts/kg and 4.63 eggs/kg. Carrots and radish were contaminated and respective numbers observed for Giardia were 155 and 59.1 cysts/kg; Ascaris was discovered in numbers of 0.7 and 1.64 eggs/kg, respectively. However, cultures irrigated with treated waste water and fresh water were free from contamination. Cysts and eggs on coriander persisted for a maximum of 8 days. PMID- 10477067 TI - Comparison of effects of Wasabia japonica and allyl isothiocyanate on the growth of four strains of Vibrio parahaemolyticus in lean and fatty tuna meat suspensions. AB - Lean tuna meat suspensions (LEAN), with a fat content of 0.006%, and fatty tuna meat suspension (FATTY), with a fat content of 3.0% were inoculated with four strains of Vibrio parahaemolyticus and wasabi (Wasabia japonica Matsumura) or allyl isothiocyanate (AIT) was added before incubation at 37 degrees C. During the incubation, viable Vibrio counts were determined on TCBS agar plates. Both LEAN and FATTY suspensions were inoculated with V. parahaemolyticus AOTO-81, (1.28+/-0.20) x 10(2) CFU/ml, followed by addition of 20 mg wasabi/ml, and incubation for 8 h. The viable Vibrio counts were (7.76+/-5.93) x 10(5) CFU/ml in LEAN and (3.50+/-2.65) x 10(1) CFU/ml in FATTY. When the same strain, at (1.18+/ 0.22) x 10(2) CFU/ml, was incubated for 8 h with 50.9 microg AIT/ml, viable Vibrio counts were (4.79+/-1.78) x 10(4) CFU/ml in LEAN and (1.80+/-1.30) x 10(1) CFU/ml in FATTY. Growth of the other three strains with wasabi or AIT was shown to be less in FATTY than in LEAN. These results indicate that growth of V. parahaemolyticus is inhibited more in FATTY than in LEAN by wasabi and allyl isothiocyanate. PMID- 10477068 TI - Combination of vaccination and competitive exclusion to prevent Salmonella colonization in chickens: experimental studies. AB - Vaccination and competitive exclusion (CE) represent accepted prophylactic measures to control Salmonella infections in chickens. To use the advantages of both the CE technique and vaccination with live Salmonella vaccines the combination of these methods was studied. In three experiments, SPF chickens were pre-treated using combined or unique administration of CE and vaccination with a live Salmonella typhimurium strain on days 1 and 2 of life and challenged with the antibiotic resistant, but otherwise isogenic mutant of this Salmonella typhimurium strain on days 3, 15 or 40 of life. The caecal colonization of both the vaccine and the challenge strain and the antibody response after infection were examined to evaluate the protective effects of the different combinations. The exclusion effect of CE cultures against Salmonella infection could be seen in very young chicks and was still considerable on day 40 of life of the birds. The Salmonella wild-type strain used as vaccine alone also resulted in a substantial protective effect against homologous challenge. The combined administration of competitive exclusion and immunization using the Salmonella wild-type strain as vaccine resulted in a considerable additional protective effect above the level of the respective exclusive application of these prophylactic measures. Administration of the Salmonella vaccine strain prior to or simultaneously with the CE culture produced the best protective effect, because such combinations ensure an adequate persistence of the vaccine strain as prerequisite for the expression of colonization inhibition effects and a strong immune response. The full exploitation of this potential using attenuated live Salmonella vaccines will require the presence of high inhibitory and immunogenic properties in the vaccine strain after attenuation of a selected parent strain. The combination of competitive exclusion and vaccination as a new measure in integrated control programmes against Salmonella infection in poultry could result in a considerable increase of protection in both very young and older chickens. PMID- 10477069 TI - Occurrence of Vibrio and other pathogenic bacteria in Mytilus galloprovincialis (mussels) harvested from Adriatic Sea, Italy. AB - Sixty-two samples of Mytilus galloprovincialis (mussels) harvested from approved shellfish waters in the Adriatic Sea were examined for the presence of Vibrio, Salmonella, Campylobacter, and verocytotoxin producing Escherichia coli. Vibrio spp. were isolated from 48.4% of samples; the species most frequently found were V. alginolyticus (32.2%) and V. vulnificus (17.7%), followed by V. cincinnatiensis (3.2%), V. parahaemolyticus (1.6%), V. fluvialis (1.6%) and V. cholerae non-O1 (1.6%). V. parahaemolyticus resulted negative to Kanagawa phenomenon and to PCR amplification of tdh gene. V. cholerae resulted negative to PCR amplification of sto gene. No Salmonella, Campylobacter, or E. coli verocytotoxin-producing strains were isolated. The results of this study suggest the potential risk of ingesting raw or undercooked mussels due to the frequent presence of potentially pathogenic Vibrio species. PMID- 10477070 TI - Inhibitory effect of seven Allium plants upon three Aspergillus species. AB - Antifungal activity and minimal fungicidal concentration (MFC) of extracts of garlic, bakeri garlic, Chinese leek, Chinese chive, scallion, onion bulb and shallot bulb against Aspergillus niger, A. flavus and A. fumigatus were examined. These Allium plants possessed antifungal activity, with garlic showing the lowest MFC. With the exception of scallion, the inhibitory effect of Allium plants against three Aspergillus species decreased with increasing incubation and heating temperature (P < 0.05). Acetic acid treatments of the extracts increased the inhibitory effect for all plants against three fungi (P < 0.05), and there was no significant difference in this effect among the three pH (2, 4, 6) treatments (P > 0.05) investigated. Acetic acid, at pH = 4, plus heat treatments of the extracts resulted in a greater inhibitory effect for all Allium plants against the three fungi than heat treatment alone (P < 0.05). Treatments of the extracts with NaCl, at concentrations of 0.2 and 0.4 M, did not affect the inhibitory effect of the plant extracts. The combination of acetic acid plus Allium plants was indicated to be an effective way to inhibit fungal growth. PMID- 10477071 TI - Modelling the overall effect of pH on the apparent heat resistance of Bacillus cereus spores. AB - A simple overall model is proposed to describe the effect of both the pH of the heating menstruum and the pH of the recovery medium on the apparent spore heat resistance of Bacillus cereus. Applied to foods making up both heating and recovery media, the model can be reduced to only two parameters. Its goodness of fit and its robustness enable it to be applied to the optimisation of heat treatments. However. further experiments should be undertaken to validate the model for other species and to determine the parameters related to reference species such as Clostridium botulinum. PMID- 10477072 TI - A predictive model for the non-thermal inactivation of Salmonella enteritidis in a food model system supplemented with a natural antimicrobial. AB - Home-made taramasalad, a traditional Greek appetizer, was inoculated with Salmonella enteritidis supplemented with different concentrations of oregano essential oil (0.0, 0.5, 1.0, 2.0% v/w) and stored at different temperatures (5, 10, 15, 20 degrees C). The product's pH was adjusted from 4.3 to 5.3 with lemon juice. At each combination of the environmental factors, the bacterial counts were modelled as a function of time in order to estimate the kinetic parameters of the pathogen. For comparison, two different models were used. A reduction of Salmonella enteritidis was observed in all cases and its death rate depended on the pH, the storage temperature and the essential oil concentration. Death responses as a function of pH, storage temperature and concentration of oregano essential oil were described using a quadratic function which was then used to predict the death of Salmonella enteritidis in home-made taramasalad of different compositions. PMID- 10477073 TI - Bacterial populations associated with a sorghum-based fermented weaning cereal. AB - Microbiological surveys, to determine the quality and safety, were conducted on 45 sorghum samples comprising dry powders (n = 15) and corresponding fermented (n = 15) and cooked fermented porridge (n = 15) samples collected from households in an informal settlement of the Gauteng Province of South Africa. Mean aerobic plate counts, Gram-negative counts and bacterial spore counts of sorghum powder samples decreased in fermented and cooked fermented porridge samples. However, mean lactic acid bacteria counts increased in fermented porridge samples, but decreased slightly in cooked fermented porridge samples. The mean pH value of sorghum powder samples decreased in fermented and cooked fermented porridge, respectively. Bacillus (B.) cereus was detected in all 15 sorghum powder samples, while Escherichia (E.) coli was detected in 53%, Clostridium perfringens in 27%, Listeria monocytogenes in 13% and Aeromonas spp., Salmonella spp., Staphylococcus aureus, Shigella spp. and Yersinia spp., each in 7% of sorghum powder samples. Of the fermented porridge samples, 40% contained B. cereus and 7% contained E. coli. None of the pathogens tested for were detected in cooked fermented porridge samples. B. cereus (53%), B. subtilis (21%), B. thuringiensis (13%), B. licheniformis (10%) and B. coagulans (3%) were identified from 120 isolates randomly selected from spore count plates of the highest dilution showing growth. PMID- 10477074 TI - Detection of small round structured viruses in artificially contaminated water using filter adsorption and reverse transcription polymerase chain reaction. AB - The small round structured viruses (SRSV) are common causes of gastroenteritis worldwide. Fecally contaminated water is an important vehicle for transmission, but detection of SRSV in environmental samples has been hampered by the lack of sensitive detection methods. The present work describes the detection of SRSV in artificially contaminated deionized water and raw drinking water. SRSV-containing fecal extracts were added to water and virus was recovered by filter adsorption elution, followed by flocculation. RNA was extracted and SRSV were detected by the use of reverse transcription polymerase chain reaction. The sensitivity of the method corresponded to a positive SRSV detection in 500 ml deionized water with an estimated concentration of 0.5-5 virus particles per ml. PMID- 10477075 TI - Genomic typing of Listeria monocytogenes strains by automated laser fluorescence analysis of amplified fragment length polymorphism fingerprint patterns. AB - The genetic relationship between isolates of Listeria monocytogenes belonging to different serotypes was determined and the suitability of automated laser fluorescent analysis (ALFA) of amplified fragment length polymorphism (AFLP) fingerprints was assessed by genomic typing of 106 L. monocytogenes isolates belonging to serotypes 1/2a, 1/2b, 1/2c, 3a, 3b, 3c, 4a, 4ab, 4b, 4c, 4d, 4e, 1, and 7. Digitised AFLP fingerprints were obtained that showed approximately 50 clearly distinguishable selectively amplified EcoRI/MseI bands for each strain. The coefficient of similarity between the profiles was determined by simple matching (Ssm). Based on these coefficients of similarity the investigated strains clustered in two genomic groups. The first group consisted of strains belonging to serotype 1/2a, 1/2c, 3a and 4a, while the second group was comprised of strains belonging to serotypes 1/2b, 3b, 4ab, 4b, 4e and 1. The average simple matching coefficient of similarity between strains of the second group was 92%, which was 4% higher than within group 1. Hence, the serotypes which are responsible for the majority of the listeriosis cases, 1/2a, 1/2b and 4b, fall into two distinct genetic groups, in concordance with their flagellar antigen type. The discriminatory power of AFLP in combination with automation of the analysis of the fingerprint profiles by ALFA makes AFLP-ALFA highly suitable for typing L. monocytogenes. PMID- 10477076 TI - Bacteriological investigation of an outbreak of Clostridium perfringens food poisoning caused by Japanese food without animal protein. AB - An outbreak of Clostridium perfringens food poisoning occurred in a senior citizen's home in Japan. Japanese food, spinach boiled with fried bean curd, was considered to be the causative food as a result of the detection of the C. perfringens enterotoxin gene by nested PCR. The number of enterotoxin-positive C. perfringens was enumerated as 4.3 x 10(5)/g in the causative food by the MPN method combined with nested PCR. By cultivation, enterotoxin-positive C. perfringens was isolated from all the fecal specimens of patients tested and the causative food. The isolates from patients were serotypable, heat-resistant and the majority produced enterotoxin, however most isolates from the causative food were nonserotypable, enterotoxin-negative and heat-sensitive. PMID- 10477077 TI - Synthesis and biological activity of a photoaffinity-biotinylated pheromone biosynthesis activating neuropeptide (PBAN) analog. AB - To study the mode of action of pheromone-biosynthesis activating neuropeptide (PBAN) at the receptor level and for receptor purification, we synthesized and tested the biologic properties of a photoaffinity biotinylated PBAN analog N-[N (4-azido-tetrafluorobenzoyl)-biocytinyloxyl-succinimide (Atf-Bct-NHS-PBAN). The Atf-Bct-NHS-PBAN was separated from unreacted reagent and synthetic Hez-PBAN by high-performance liquid chromatography. Conjugated biotin was detected by using enzyme-linked assay as well as tricine sodium dodecyl sulfate polyacrylamide gel electrophoresis. The biologic activity of purified Atf-Bct-NHS-PBAN was confirmed using both in vivo and in vitro pheromonotropic bioassays. These observations indicate that Atf-Bct-NHS-PBAN is a full agonist of PBAN action in pheromone glands and may be used to study PBAN receptors by employing avidin coupled to various reporter groups. PMID- 10477078 TI - Structure-activity studies on cysteine-substituted neurokinin A analogs. AB - A complete series of analogs of tyrosine modified neurokinin A ([Tyr1]-NKA or [Tyr0]-NKA) has been synthesized by substituting each natural residue with 1-Cys. These analogs were tested for their ability to bind recombinant neurokinin-2 (NK 2) receptor. Substitution of Phe6 with Cys completely abolished binding of the analog to the receptor. Substitution of residues in the carboxyl-terminal region of the peptide (Met10, Leu9, Gly8, Val7) and Asp4 with Cys gave reductions in binding affinity of between 23- and 250-fold. Molecular dynamics simulations of these analogs suggest that changes in peptide structure and flexibility are not large contributors to the losses in receptor binding affinity. Reductions in binding affinity are therefore more confidently ascribed to losses of peptide receptor interactions. PMID- 10477079 TI - Alteration of neurotensin receptors in MPTP-treated mice. AB - We examined the sequential changes in neurotensin receptors in the striatum and substantia nigra of mouse brains lesioned with 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) by receptor autoradiography, in comparison with the alterations in dopamine uptake sites. The mice received four intraperitoneal injections of MPTP (10 mg/kg) at 1-h intervals and then the brains were analyzed at 6 h and 1, 3, 7, and 21 days after the treatments. [3H]Neurotensin and [3H]mazindol were used to label neurotensin receptors and dopamine uptake sites, respectively. [3H]Neurotensin binding was significantly decreased in the striatum from 6 h to 21 days after MPTP treatment. In the substantia nigra, pars reticulata also showed a significant decrease in [3H]neurotensin binding from 3 to 21 days post-MPTP treatment. However, no significant change in [3H]neurotensin binding was observed in the pars compacta even after 21 days. On the other hand, [3H]mazindol binding was markedly decreased in the striatum and substantia nigra from 6 h to 21 days after MPTP treatment. These results indicate that neurotoxin MPTP can produce a severe decrease in neurotensin receptors and dopamine uptake sites in the striatum and substantia nigra of mice. Thus, our findings provide evidence that the dysfunction in neurotensin receptors may be involved in the degenerative processes causing Parkinson's disease. PMID- 10477080 TI - Neuropeptide Y release by pumiliotoxin-B in the electrically-stimulated mouse vas deferens: an immunohistochemical study. AB - Morphologic and immunohistochemical studies were conducted to ascertain whether pumiliotoxin-B (PTX-B), an indolizine alkaloid from the skin of the Neotropical dendrobatid frog, Dendrobates pumilio, affects the anatomic and immunohistochemical features of the electrically stimulated mouse vas deferens preparations. PTX-B, at a concentration of 1 microM, consistently decreased the density pattern of neuropeptide Y (NPY)-immunoreactive nerve fibers contained within the circular muscular layer. The alkaloid also induced striking morphologic changes. It enlarged the lumen of the vasa and relaxed the muscular wall. Pretreatment with prazosin or haloperidol affected neither the release of NPY nor the morphologic changes; pretreatment with tetrodotoxin and guanethidine abolished NPY release and prevented the PTX-B-induced morphologic changes. PTX-B had no appreciable effect on the density and distribution pattern of nerve fibers immunostained for vasoactive intestinal polypeptide, substance P, calcitonin gene related peptide, enkephalin, pancreatic polypeptide, 5-hydroxy-tryptamine and tyrosine hydroxylase. PMID- 10477081 TI - Differential involvement of PKA and PKC in regulation of catecholamine enzyme genes by PACAP. AB - Roles of protein kinase A (PKA) and protein kinase C (PKC) in regulation of tyrosine hydroxylase, dopamine beta-hydroxylase, and phenylethanolamine N methyltransferase expression by pituitary adenylate cyclase-activating polypeptide (PACAP) were determined in primary cultured bovine chromaffin cells. DBH up-regulation by PACAP was reduced by H-89 and not further increased by forskolin showing involvement of cAMP/PKA. It was not mediated by PKC, as 12-O tetradecanoylphorbol-13-acetate and sphingosine exerted no effect. Tyrosine hydroxylase induction by PACAP was mediated by both kinases. The PACAP-activated PKA up-regulated phenylethanolamine N-methyltransferase expression whereas PKC caused down-regulation. PACAP increased tyrosine hydroxylase and dopamine beta hydroxylase activities, but slightly lowered phenylethanolamine N methyltransferase activity, resulting in a preferential rise in norepinephrine over epinephrine. PMID- 10477082 TI - Gluconeogenesis activation after intravenous angiotensin II in freely moving rats. AB - Intravenous (IV) administration of angiotensin II (0.95 nmol/100 g body weight) produced a marked increase in plasma glucose of 20 h fasted rats. To investigate the possibility of a stimulation of gluconeogenesis, conscious unrestrained rats were continuously infused with [14C]bicarbonate, 60 microl/min (0.18 microCi/min), and label incorporation into circulating glucose was determined before and after angiotensin injection. The rate of 14C incorporation into blood glucose of fed rats increased significantly after angiotensin II administration, a 279% increase after 20 min (P < 0.01). In conclusion, the results of the present study show that the hyperglycemia induced by intravenous (IV) administration of angiotensin II is accompanied by an activation of gluconeogenesis, as evidenced by a rapid and marked increase in the rate of 14CO2 incorporation into circulating glucose. PMID- 10477083 TI - Characterization of a functional AT1A angiotensin receptor in pancreatoma AR4-2J cells. AB - Functional angiotensin receptors were characterized in the rat pancreatic acinar cell line AR4-2J. Angiotensin II stimulated a dose-dependent release of amylase and production of inositol phosphates. Results of high-performance liquid chromatography separation of inositol phosphates indicated that angiotensin stimulated the rapid accumulation of inositol 1,3,4-trisphosphate. Angiotensin II and angiotensin III were at least an order of magnitude more potent than angiotensin I in the stimulation of amylase release. The angiotensin II stimulated amylase release was blocked by losartan, a selective AT1 angiotensin antagonist. The selective AT2 angiotensin receptor ligands CGP42112 did not alter angiotensin II-stimulated amylase released. However, CGP42112 stimulated amylase release at micromolar concentrations with a potency similar to angiotensin I. Analysis of mRNA expression by reverse transcription polymerase chain reaction suggested that AT1A was the predominant type-I angiotensin receptor expressed in the AR4-2J cells. PMID- 10477084 TI - MCF-7 breast cancer cells express normal forms of all vasopressin receptors plus an abnormal V2R. AB - We have previously provided evidence that an autocrine loop involving vasopressin is present in perhaps all breast cancers. This study now shows MCF-7 breast cancer cells express mRNAs for all currently recognized vasopressin receptor subtypes (V1a, V1b, and V2). Cloning and DNA sequencing over the entire open reading frame of each mRNA revealed that normal sequences representing each receptor were present. However, in addition, an abnormal mRNA for the V2 receptor, expected to give rise to a truncated 'diabetic' protein, was also expressed. Western analysis revealed that all three normal mRNAs gave rise to proteins of sizes compatible with them being functional receptors. The abnormal V2 receptor mRNA also gave rise to proteins. PMID- 10477085 TI - Antagonistic analogs of growth hormone releasing hormone (GHRH) inhibit cyclic AMP production of human cancer cell lines in vitro. AB - Antagonistic analogs of growth hormone-releasing hormone (GHRH) inhibit growth of various human cancers both in vivo and in vitro. GHRH, vasoactive intestinal peptide (VIP), and pituitary adenylate cyclase-activating peptide stimulate cyclic AMP (cAMP) release from various human cancer cell lines in vitro. Thus, in the present study, we investigated the effects of antagonistic analogs of GHRH on the GHRH- and VIP-induced cAMP release from cultured human cancer cells in a superfusion system. Various human cancer cell lines were exposed to human GHRH(1 29)NH2 (2-20 nM) or VIP (0.1-5 nM) repeatedly for 12 min or continuously for 96 min. GHRH antagonist MZ-5-156 at 100 to 200 nM concentration inhibited the GHRH- or VIP-induced cAMP release from mammary (MDA-MB-468), prostatic (PC-3), and pancreatic (SW-1990 and CAPAN-2) cancer cells. These results show that antagonistic analogs of GHRH suppress the stimulatory effects of GHRH and VIP on the cAMP production of various cancer cells. Because cAMP is a potent second messenger controlling many intracellular functions, including the stimulation of cell growth, an inhibition of autocrine/paracrine action of GHRH by the GHRH antagonists may provide the basis for the development of new methods for cancer treatment. PMID- 10477086 TI - Intracisternal sauvagine is more potent than corticotropin-releasing factor to decrease gastric vagal efferent activity in rats. AB - Consecutive intracisternal (ic) injections of corticotropin-releasing factor (CRF) (21, 63, and 126 pmol, ic) or sauvagine (2.1, 6.3, and 21 pmol, ic) decreased gastric vagal efferent multiunit discharge (GVED) to 82%, 75% and 69% and 71%, 40% and 21%, respectively, from preinjection basal levels (taken as 100%). The inhibitory action was dose related (magnitude and duration of the response, 7-45 min). The CRF antagonist, [D-Phe12,Nle21,38,Calpha-MeLeu37]-rCRF12 4 1 (6.25 nmol, ic) increased GVED by 43.5+/-4.3% and blocked the decrease in GVED induced by CRF (21 pmol, ic) for >90 min with a complete recovery after 3 h. Vehicles (injected intracisternally) had no effect. These data indicate that: 1) CRF injected intracisternally decreases GVED through the activation of CRF receptors and sauvagine is more potent than CRF to inhibit GVED; and 2) endogenous CRF exerts an inhibitory tone on basal GVED in urethane-anesthetized rats undergoing surgery. PMID- 10477087 TI - Thrombin-stimulated increases in cytosolic Ca2+ level and gonadotropin-releasing hormone release in GT1-7 neurons. AB - The effects of thrombin on cytosolic calcium levels ([Ca2+]cyt), and on gonadotropin-releasing hormone (GnRH) release, were characterized in cultured GT1 7 neurons. GnRH release from GT1-7 neurons was pulsatile with an average pulse amplitude of 14.3+/-5.8 pg x min x ml(-1) and an average pulse duration of 21.3+/ 4.2 min. The [Ca2+]cyt response to 0.005 to 0.2 U/ml thrombin was saturable and concentration dependent (EC50 = 0.0268 U/ml). Ethyleneglycotetraacetic acid (EGTA) chelation of extracellular Ca2+ resulted in an approximately 70% attenuation of thrombin-stimulated increase in [Ca2+]cyt. By use of a special superfusion system, a 5-min exposure to 0.1 U/ml thrombin significantly increased the amplitude (193.2+/-67.8 pg x min x ml(-1); P = 0.001) but not the duration (22.5+/-2.4 min; P = 0.8) of GnRH release. These results suggest that thrombin increases [Ca2+]cyt and GnRH release from GT1-7 neurons via specific membrane bound receptors. PMID- 10477088 TI - The proenkephalin A-processing product peptide E, which encompasses two enkephalin sequences, has a much lower opioid activity than beta-endorphin. AB - Peptide E is a 25-amino acid peptide derived from proenkephalin A that was originally isolated from the bovine adrenal medulla. Bovine peptide E (BPE), which possesses a Met- and a Leu-enkephalin sequence at its N- and C-terminus, respectively, has been described as a highly potent and selective mu-opioid receptor agonist. Paradoxically, the frog counterpart of peptide E (FPE), which exhibits only two amino acid substitutions (Met15-->Gln and Leu25-->Met) compared with BPE, was found to be totally devoid of antinociceptive activity. To decipher this apparent discrepancy, we have decided to compare the structural and pharmacological characteristics of FPE, BPE, and the chimeric peptide [Gln15]BPE (Q15BPE). In methanol, all three peptides exhibited virtually the same conformation, the central region of each peptide (residues 10-20) being involved in a regular helix. Intracerebroventricular administration of FPE, BPE, or Q15BPE, at doses up to 1000 ng per mouse, did not induce any analgesic effects, as evaluated by the hot plate and writhing tests, whereas, in the same tests, beta-endorphin at a dose of 100 ng provoked profound analgesia. Concomitant administration of FPE, BPE, or Q15BPE (100 ng) with the aminopeptidase-N inhibitor bestatin (50 microg) or the endopeptidase 24-11 inhibitor thiorphan (10 microg) did not produce analgesic responses. Antinociceptive effects were only observed when very high doses of FPE, BPE, and Q15BPE (10000 ng per mouse) were administered. These data clearly demonstrate that, contrary to what has been previously reported, peptide E is virtually devoid of opioid activity. PMID- 10477089 TI - Electron microscopic observation of mu-opioid receptor in the rat area postrema. AB - A simple preembedding avidin-biotin-peroxidase complex technique was used to study the ultrastructural localization of mu-opioid receptor in the rat area postrema. By using low concentrations of the first antiserum for incubation with a short reaction time to 3,3'-diaminobenzidine, the immunostaining was faint at the light microscopic level. However, at the electron microscopic level, strong immunoreaction was observed. Mu-Opioid receptors were found to be localized on the postsynaptic membrane of dendrites, extrasynaptic plasma membrane, and the surface of the small, clear vesicles in axon terminals. Of the total 283 immunopositive profiles observed, 68.2% (193 of 283) were dendrites, 29.3% (83 of 283) were axon terminals, and 2.5% (7 of 283) were myelinated axons. No immunostained neuron bodies were found in the present study; 109 mu-opioid receptor immunoreactive dendrites received synapses (56.5%, 109 of 193) from nonimmunoreactive (84.4%, 92 of 109) or immunoreactive (15.6%, 17 of 109) axon terminals, whereas 84 dendrites (43.5%, 84 of 193) were found without receiving synapses. The present study shows that the mu-opioid receptor in the area postrema plays a role mainly at the synapses. PMID- 10477090 TI - 2,2'-Bispyridyl disulfide rapidly induces intramolecular disulfide bonds in peptides. AB - A linear peptide containing two reduced cysteine residues can be rapidly converted to its oxidized cyclic form containing an intramolecular disulfide bond by adding an excess of 2,2'-bispyridyl disulfide (2,2'-dipyridyl disulfide or 2,2'-dithiodipyridine) to conventional buffer solutions. The reactants and products are easily separated by reverse-phase chromatography. This reaction will find wide application in forming intramolecular disulfide bonds because of its selectivity for free sulfhydryl groups, quickness, safety, and applicability under acidic conditions. PMID- 10477091 TI - Signaling pathways mediating gastrin's growth-promoting effects. AB - In addition to its fundamental role in stimulating gastric acid secretion, the peptide hormone gastrin induces growth-promoting effects on diversity of target cells. Various mechanisms, including endocrine, paracrine, and autocrine, have been proposed for gastrin's growth-promoting actions. The mitogenic effects of gastrin are mediated by specific cell surface receptors activated after gastrin binding. The functionally defined receptors for gastrin include cholecystokinin A (CCKA) receptor, which is discriminating for sulfated CCK8; cholecystokinin B (CCKB)/gastrin receptor, which binds gastrin17 sulfated, and nonsulfated CCK8 with nearly equal affinities; cholecystokinin C (CCKC), which is a low-affinity gastrin binding protein; and novel, high-affinity receptors selective for amidated gastrin, processing intermediates of gastrin, or both. The signaling pathways mediating gastrin's stimulation of the CCKB/gastrin receptor have been progressively outlined, and the pathways mediating other receptors have been slowly emerging. Engagement of the gastrin receptor initiates various biochemical and molecular events, including recruitment and activation of tyrosine kinases, activation of the phospholipase C signaling pathway leading to phosphoinositide breakdown, intracellular calcium mobilization and protein kinase C stimulation, activation of the mitogen-activated protein kinase pathway, and induction of early response genes. Current emphasis is on understanding the functional significance of processing intermediate forms of gastrin, and the receptor subtypes and pathways that promote the trophic/mitogenic effects of the different molecular forms of gastrin. PMID- 10477092 TI - Full-size digitized mammography will probably be a major step forward in breast imaging. PMID- 10477093 TI - Full field digital mammography scanner. AB - We describe the key features of a pre-production, slot-scan digital mammography system. A number of these units have been used in clinical studies over the past year for the purpose of demonstrating their equivalence to the conventional film screen devices. Since the clinical data has not yet been fully analyzed, it is not possible to make definitive claims. However, with hundreds of patients examined, the results appear to leave very little doubt the SenoScan digital mammography system will prove equivalent to the conventional technology. The detector developed for this system has a sensitive area 1.0 cm wide by 22 cm long. It is constructed by abutting four charge-coupled-device (CCD) chips, which are optically coupled to thallium-doped cesium iodide scintillator by means of a thin fiber optic plate. Scanning is accomplished by attaching the detector to a rigid arm that swings in an arc, with the axis of rotation collinear with the X ray tube focal spot. The total scan time for the 30 cm image width is less than 6 s, with an effective exposure time of either 0.2 or 0.4 s. Two resolution modes are available: 0.054 mm or 0.027 mm square pixel size; in the latter mode both the image length and width are halved, as is the scan velocity, so that the scan time remains the same. To compensate for the low X-ray utilization efficiency of the slot geometry, a tungsten rhenium target X-ray tube is employed. It is rated at 8 kW on the 0.3 mm focal spot; when used with a heat exchanger, it has been found to provide the patient throughput needed in a busy clinical practice. PMID- 10477094 TI - Digital mammography performed with computed radiography technology. AB - Introduced by Fuji Photo Film Japan in the early 1980s, computed radiography (CR) technology has developed considerably since then to become the mature widely installed technology it is today (about 7500 systems worldwide). Various mammographic examinations require high performance results to which CR complies on demand or following some procedures such as geometrical magnification carried out during the examination. The basic CR principles and digital image processing as well as technical improvements are detailed in this study, which also includes a synthesis of the articles on CR mammographic applications referenced in the bibliography, focusing on strong points, limits and current methods of surpassing these limits. New CR technology development perspectives in mammography and computed assisted diagnosis (CAD) algorithms will allow wider use of this method in the near future. PMID- 10477095 TI - Full-field digital mammography designed as a complete system. AB - Although mammography is currently considered by many to be the best tool for the early detection of breast cancer, conventional film-screen imaging is still far from perfect. A fundamental limitation of film-screen mammography is the fact that the detection, display and storage devices are one and the same, making it impossible to separately optimize each device. In addition to a potentially equivalent or better image quality compared to the film-screen technology, full field digital mammography (FFDM) will provide the radiologist with numerous advantages: digital image management, digital data transfer and new medical applications. Nevertheless, to fulfil the potential of digital mammography, an FFDM unit must not only provide an outstanding technology for X-ray detection, but should be designed as a complete system. Each component of the FFDM system should be designed to provide or to increase the current image quality, the current ease of use and the current throughput: the X-ray tube, the gantry, the image receptor packaging, the detector, the acquisition electronics, the softcopy review workstation, the user interface, as well as the network capabilities. PMID- 10477096 TI - The promise of computer aided detection in digital mammography. AB - Computer-aided detection (CAD) is a new technology now being implemented in many clinics to reduce the false negative rate in mammography screening. A large clinical study has been completed which shows that a substantial false negative (miss) rate exists in screening mammography, a significant fraction of the missed cancers are not subtle, and CAD has high sensitivity to these missed cancers. Full field digital mammography is now coming on the scene, but has not yet been proven in clinical practice. The authors believe that full acceptance of the new digital technology depends not merely on demonstrations of 'substantial equivalence' to film-screen technology, but rather on more complete exploitation of the unique advantages of digital technology, and that CAD can play a key role. These advantages derive from CAD's ability to quickly (in near real-time) perform analytical computations on digital information that is not readily available to the radiologist until after the cost of film-processing has occurred. PMID- 10477097 TI - Quiz case 9. Breast emphysema. PMID- 10477098 TI - Spiral CT of the abdomen: increased diagnostic potential. AB - Over the last 10 years, spiral CT has become the optimal method of performing all CT within the abdomen and pelvis. Not only has the technique improved the quality of individual examinations, it has also opened up many new diagnostic possibilities which are currently being evaluated and compared with the concurrent advances in ultrasound and MRI. These new diagnostic possibilities are discussed in this review article. So too are the economic advantages of the shorter data-acquisition times which also make the examination much better tolerated by the patient. PMID- 10477099 TI - A rhinolith which is mimicking a nasal benign tumor. PMID- 10477100 TI - Thrombocytopenic thrombotic purpura: severe clinic with no CT, minor MRI, but a SPECT correlate. AB - A 28-year-old woman with primarily therapy refractory TTP was followed neuroradiologically over 6 months. Despite pronounced neurological and neuropsychiatric symptoms including hemiparesis and aphasia she had unremarkable CT scans on two occasions. Three MRI exams showed no correlate for her neurological symptoms except a small petechial cortical hemorrhage in the right parietooccipital gyrus which may account for her TTP-related anxiety disorder. A cerebral HMPAO-SPECT showed long-standing right-sided hypoperfusion compatible with residual vasculature changes. The possible causes for the clinico neuroradiological discrepancies are discussed in view of the literature. PMID- 10477101 TI - Negative captopril renography on patients with renin mediated hypertension due to page kidney and reninoma. AB - Through a mechanism similar to renal artery stenosis, patients with reninoma and page kidney also suffered from renin mediated hypertension. Captopril renograms performed on our patients with the latter two conditions, however, did not yield diagnostic findings. Therefore, equivocal or negative captopril renography cannot serve to rule out conditions with elevated renin other than renal artery stenosis. PMID- 10477102 TI - Intracranial meningiomas: correlations between MR imaging and histology. AB - The authors have examined the relationship between magnetic resonance imaging (MRI) and histopathological features in 35 surgically verified intracranial meningiomas. Tumor signals on T1-weighted images were rather similar regardless of the histologic subtype of the tumors. On T2-weighted images, hypointense meningiomas were mainly fibroblastic and hyperintense tumors were mainly syncytial and angioblastic, and partly transitional. Isointense tumors were mainly transitional and partly fibroblastic and syncytial. The authors conclude that the signal intensity of the MRI may be useful in the preoperative characterization of intracranial meningiomas. T1-weighted images may predict the presence of cysts and intratumoral blood vessels; whereas T2-weighted images can give information about histological subtype, vascularity and consistency. Meningiomas hyperintense to the cortex on T2 are usually soft, more vascular and more frequently of syncytial or angioblastic subtype; tumors hypointense or hypo isointense on T2 tend to have a more hard consistency and are more often of fibroblastic or transitional subtype. PMID- 10477103 TI - Permeability of iodinated and MR contrast media through two types of hemodialysis membrane. AB - The clearance of three iodinated contrast media (CM) and three MR-CM through two kinds of hemodialysis (HD) membranes were investigated in vitro. All three MR-CM are not only injected intravenously and mainly secreted through the kidney, but also now commercially available. Each of the six CM showed significantly higher clearance with one kind of HD membrane, with a larger pore size, than that with the other. There were also large differences in clearance between the three iodinated CM with both kinds of HD membranes, but not between the three MR-CM with either kind of membrane. Thus, in order that iodinated CM be removed from the body as soon as possible in HD patients, it is important to consider the choice not only among the iodinated CM, but also between the type of HD membrane that is used. Concerning MR-CM, only the choice of HD membrane is important. PMID- 10477104 TI - Age-related decrease in brain-derived neurotrophic factor gene expression in the brain of the zitter rat with genetic spongiform encephalopathy. AB - The zitter rat exhibits a progressive degradation in neuronal cells and genetic spongiform encephalopathy with age. In order to elucidate the involvement of the expression of the neurotrophic factor in neuropathology of the rat, we quantified mRNA levels of neurotrophic factors (nerve growth factor, brain-derived neurotrophic factor (BDNF), and neurotrophin-3, ciliary neurotrophic factor and glial-cell-line-derived neurotrophic factor) in the zitter rat brain. Expression of the BDNF gene was lower in the zitter rat brain (cerebrum, cerebellum, and brainstem regions). Interestingly, kinase activity of mitogen activated protein kinase (MAPK) Erk2 involved in the expression of BDNF was also down regulated, despite an unchanging expression of MAPK protein. These results show the possible involvement of a MAPK pathway in BDNF mRNA reduction in the zitter rat brain. PMID- 10477105 TI - Blockade of dopamine D2, but not of D1 receptors in the rat globus pallidus induced Fos-like immunoreactivity in the caudate-putamen, substantia nigra and entopeduncular nucleus. AB - In the present study, we investigated Fos-like immunoreactivity (FLI) evoked by pallidal dopamine (DA) D1 and D2 receptor blockade in the caudate-putamen (CPu), substantia nigra (SN) and entopeduncular nucleus (EP), i.e. major target areas of pallidal efferents. Results demonstrate that infusion of the selective D1 antagonist SCH23390 (1 and 4 microg/0.5 microl) into the globus pallidus (GP) did not induce FLI in the CPu, SN and EP. In contrast, intrapallidal infusion of a low dose of the selective D2 antagonist S(-)-sulpiride (15 microg/0.5 microl) induced FLI restricted to the CPu. A higher dose of intrapallidal S(-)-sulpiride (25 microg/0.5 microl) induced FLI in the CPu as well as in the SN and EP. These findings add further evidence to notion that the GP plays a central role in the basal ganglia circuitry and demonstrate an involvement of extrastriatal DA via D2 receptors. PMID- 10477106 TI - Free-running circadian period does not shorten with age in female Syrian hamsters. AB - It has been reported that the free-running period of circadian rhythms shortens with age in mammals, including humans, and this shortening has been suggested to be the underlying cause of early morning awakening and difficulty maintaining sleep in older people. A recent study found that the free-running period of male hamsters does not change with age. The present study extends those findings to female hamsters. We studied the locomotor activity rhythm of 22 female hamsters kept in constant conditions from early adulthood until their death, and compared their data to those from male hamsters. We found no shortening of free-running period with age in the female hamsters, and no difference in free-running period between females and males. In contrast, mean activity level and amount of time per cycle spent running declined with age in females and males. These findings demonstrate that the free-running period in hamsters does not systematically shorten with age, and suggest that alternative explanations for the observed age related advance of sleep-wake times in humans should be explored. PMID- 10477107 TI - Transient upregulation of osteopontin mRNA in hippocampus and striatum following global forebrain ischemia in rats. AB - We investigated the spatial and temporal expression of osteopontin (OPN) mRNA following transient forebrain ischemia in rats. Experiments were carried out using a four-vessel occlusion model for forebrain ischemia. The transient induction of OPN mRNA after global ischemia occurred earlier in the striatum than in the hippocampus. It was pronounced in the dorsomedial striatum close to the lateral ventricle and in the CA1 subfield and the subiculum of the hippocampus before microglial cells became more reactive. It also could be detected in the dentate hilus and to a marginal extent in the CA3. Our results suggest that the hippocampus and the striatum following global forebrain ischemia upregulate OPN mRNA in different spatiotemporal profiles. PMID- 10477108 TI - Left hemisphere activation during processing of morphologically complex word forms in adults. AB - Functional neuroanatomy of the processing of morphologically complex words was studied by measuring regional brain activity by positron emission tomography (PET) during encoding of auditorily presented inflected versus monomorphemic Finnish nouns. Significant increases of activation occurred particularly in the left inferior posterior frontal lobe, corresponding to Broca's area. This suggests that besides their role in the production of grammatical morphology documented earlier, Broca's area and adjacent regions are important for the input processing of morphologically complex words. PMID- 10477109 TI - Cortical asymmetries of the human somatosensory hand representation in right- and left-handers. AB - Hemispheric asymmetry is known for higher brain functions like language and attention. We tested whether such an asymmetry also exists in the representation of elementary sensory functions. Magnetic source imaging was used to compare the cortical somatosensory hand representation in seven right- and five left-handed individuals. In all right-handers the representation of the dominant hand was larger than the contralateral one in the corresponding hemispheres. In contrast, only two out of five left-handers revealed a larger representation of the dominant left hand compared to the right one. In agreement with previous findings on the lateralization of language and attention, there is a strong correlation between handedness and the extent of the cortical hand representation in right-, but not in left-handers. We conclude that a profound functional hemispheric asymmetry also exists in primary sensory cortices. PMID- 10477110 TI - Accumulation of mercury in neurosecretory neurons of mice after long-term exposure to oral mercuric chloride. AB - Inorganic mercury (HgCl2) was administered to adult mice in drinking water (20 mg/l). Animals were sacrificed after one or two years and fixed by whole-body perfusion. Sections of the hypothalamus and neurohypophysis were subjected to silver acetate autometallography for visualization of mercury at light and electron microscopy levels. Mercury deposits, which can be seen by light microscopy as black granules, were found to accumulate within neuronal perykaria of the supraoptic and paraventricular nuclei. Electron microscopy demonstrated that mercury deposits in neurosecretory neurons were detected exclusively within lysosomes. Mercury was also present in small vesicles, 40-70 nm in diameter, and in endocytic vacuoles within the axon terminals of the neurohypophysis. No mercury could be seen in sections obtained from control animals that had been drinking uncontaminated water. Mechanisms involved in uptake and transport of mercury to neuronal bodies are discussed. PMID- 10477112 TI - Differential activation of c-Jun N-terminal protein kinase and p38 in rat hippocampus and cerebellum after electroconvulsive shock. AB - Electroconvulsive shock (ECS), an effective treatment for psychiatric diseases, has been reported to induce immediate-early genes (IEGs) and to activate p42 and p44 MAPKs (ERK-1 and ERK-2) in rat brain. In this study, we examined the activation of the other members of MAPK family, c-Jun N-terminal protein kinase (JNK/SAPK) and p38. Following ECS, the phosphorylation of p38 was substantially increased in both hippocampus and cerebellum, but the increase of JNK phosphorylation was observed only in hippocampus. We also investigated the phosphorylation of their upstream kinases, SEK-1, MKK6 and MKK3. In both hippocampus and cerebellum, the phosphorylation of MKK6 showed closer correlation with p38 phosphorylation than that of MKK3. However, SEK-1, known as upstream kinase of JNK and p38 in vitro, corresponded with none of MAPKs. These results, with previous reports on the activation of ERK, indicate that ECS activates three MAPKs differentially in rat hippocampus and cerebellum, and suggest the possibility that unknown MAPKK may be involved in the activation of JNK in rat brain after ECS. PMID- 10477111 TI - Regulated secretion of amyloid precursor protein by TrkA receptor stimulation in rat pheochromocytoma-12 cells is mitogen activated protein kinase sensitive. AB - We have shown recently in the pheochromocytoma PC-12 cell line, that the activation of the high-affinity receptor for nerve growth factor (NGF), tyrosine kinase receptor (TrkA), results in increased secretion of the amyloid precursor protein (APP) into the culture medium. In order to reveal through which TrkA associated signaling pathway the secretory APP processing is mediated, signaling cascades activated by TrkA stimulation were selectively inhibited under conditions of selective TrkA stimulation via non-NGF mechanisms and APP secretion into the culture medium was followed by Western analysis. Our data demonstrate, that activation of mitogen activated protein (MAP) kinase alone is sufficient to promote APP secretion, whereas inhibition of MAP kinase will reduce APP secretion only when phospholipase Cgamma or phosphatidylinositol 3-kinase are additionally inhibited. This suggests that pharmacological manipulations activating the MAP kinase pathway may result in increased secretory APP processing. PMID- 10477113 TI - Enhanced Fos expression in the hippocampus of El mice after short-term vestibular stimulation. AB - The El mouse is an animal model for human epilepsy. The mouse manifest seizures in response to periodically repeated vestibular stimuli such as being tossing-up. Although this technique has been traditionally employed to accelerate this disease in the mouse, its meaning remained obscure. The present study was conducted to estimate the effects of tossing-up stimuli on expression of c-fos, a well-known marker of neuronal activation. Expression of c-fos was significantly increased even after single tossing-up specimen. It was blocked by pretreatment of the mouse with MK-801, a NMDA receptor antagonist. A marked expression of the oncoprotein was observed in the granule cell layer of dentate gyrus and CA1-3 pyramidal cell layer in the hippocampus of the mouse. These results suggest that the El mice are genetically hyper-sensitive to the vestibular stimuli. PMID- 10477114 TI - Expression of inducible nitric oxide synthase immunoreactivity in rat brain following chronic hypoxia: effect of aminoguanidine. AB - To clarify the role of inducible nitric oxide synthase (iNOS) in the histopathological changes that occur in the brain after exposure of rats to normobaric hypoxia (10% O2 in N2) for 2 weeks, we examined the localization of iNOS and the effect of aminoguanidine, a relatively selective iNOS inhibitor, on the histological outcome. Animals were divided into a hypoxia group, an aminoguanidine-treated hypoxia group and a normoxic control group. The hypoxia group showed severe ischemic changes and prominent angiogenesis in the CA1 hippocampus and cerebral cortex. Aminoguanidine significantly reduced the ischemic change and angiogenesis in these regions, and also reduced iNOS immunoreactive cells compared to the hypoxia group. These findings suggest that iNOS activity could play a role in the neuropathological alterations induced by chronic hypoxia. PMID- 10477115 TI - Expanded expression of heme oxygenase-1 (HO-1) in the hypothalamic median eminence of aged as compared with young rats: an immunocytochemical study. AB - This study was performed to examine the differences in expression of heme oxygenase protein with age using immunocytochemistry. We compared the contents of HO-1 and HO-2 between young and aged rats using immunocytochemical methods. Stronger HO-1 expression was detected in the internal layer of the median eminence (ME) of aged than of young rats. Moreover, the cells expressing HO-1 were larger in the aged than the young animals. Electron microscopy indicated these cells with HO-1-like immunoreactivity (HO-1-LI) to be astrocytes. These findings suggested that the expression of HO-1 increased in the ME with age. The significance of this increased expression of HO-1 with age will be discussed briefly. PMID- 10477116 TI - Interaction of kyotorphin and brain peptide transporter in synaptosomes prepared from rat cerebellum: implication of high affinity type H+/peptide transporter PEPT2 mediated transport system. AB - High-affinity type H+/peptide cotransporter PEPT2 is preferentially expressed in the kidney, and is responsible for reabsorption of di- and tripeptides in epithelial tubules. Interestingly, PEPT2 has been recently cloned from rat brain. However, there is very little information available on the peptide transporter activity in the brain. In the present study, we investigated the interaction of kyotorphin (L-tyrosyl-L-arginine) with the peptide transporter using synaptosomes prepared from rat cerebellum. The activity of the peptide transporter was assessed by measuring the uptake of radiolabeled glycyl-sarcosine (Gly-Sar), which is a prototypical substrate for the peptide transporter, in the presence of H+-gradient. Kyotorphin competitively inhibited the uptake of Gly-Sar with an inhibitory constant (Ki) of 30 +/- 4 microM in rat cerebellum synaptosomes. This uptake property is very close to that of PEPT2. Carnosine (beta-alanyl-L histidine) also inhibited the uptake of Gly-Sar, on the other hand, TRH did not interact with the peptide transporter. RT-PCR using specific primers revealed that PEPT2 mRNA exists in cerebellum in rat. Taken collectively, these results indicate that the functional peptide transport system in rat cerebellum might be the high affinity transporter PEPT2. PMID- 10477117 TI - Opposite regulation by glucocorticoids of the alpha 1B- and alpha 2A adrenoreceptor mRNA levels in rat cultured anterior hypothalamic slices. AB - In this study we investigated whether the expression of alpha1B- and alpha2A adrenoreceptor mRNAs is differently modulated by glucocorticoids in rat cultured anterior hypothalamus slices. Using a semi-quantitative reverse transcription polymerase chain reaction assay, the level of the alpha1B-adrenoreceptor mRNA was significantly reduced in slices cultured in steroid free-medium when compared with that measured in standard medium (i.e. containing basal adrenosteroid plasma concentrations). In contrast, the expression of the alpha2A-adrenoreceptor mRNA was markedly increased. Finally, the ratio of alpha1B- versus alpha2A-mRNA levels was about 1.7 and 0.7 in standard and steroid-free medium, respectively. These responses were completely reversed by supplementation with corticosterone. These findings provide the first evidence that in vitro glucocorticoids may regulate, in an opposite manner, the expression of the alpha1B-and alpha2A-adrenoreceptor mRNAs in the hypothalamus. This kind of regulation could be related to steroid dependent changes in the noradrenergic control of neuroendocrine secretions. PMID- 10477118 TI - Evidence for activation of microglia in patients with psychiatric illnesses. AB - Activation of microglia/macrophages is a key event in response to pathological changes in the CNS. HLA-DR is a valuable immunohistochemical marker that specifically reacts with activated microglia cells. In order to elucidate a potential role of microgliosis in severe psychiatric illnesses, post-mortem frontal cortex and hippocampus of patients with schizophrenia (n = 14) and affective disorder (n = 6) and control specimens (n = 13) were studied. Additionally Alzheimer's disease cases (n = 8) were included as a human model system with typical neurodegenerative alterations and microglia activation. All patient groups revealed subjects with abundant microglia immunostaining (schizophrenia, three patients; affective disorder, one patient; Alzheimer's disease, four patients) in both gray and white matter. This finding provides evidence for distinct neuropathological changes in brains of patients with schizophrenia and affective disorder. The activation of microglia cells, which represent a major part of the brain immune response, may help to unravel the pathophysiological processes in severe psychiatric illnesses. PMID- 10477119 TI - Alpha-2 macroglobulin gene in early- and late-onset Alzheimer disease. AB - Alpha-2-macroglobulin (A2M) is a proteinase inhibitor that is present in senile plaques and may play a role in metabolism of amyloid beta (A beta) peptide. Recently it was reported that inheritance of the deletion allele (A2M-2) confers increased risk for late-onset Alzheimer disease (AD) with significance of this effect similar to the epsilon4 allele of apolipoprotein E (APOE). We examined the distribution of A2M genotypes and alleles in a cohort of 146 AD patients and 160 age-matched non-demented individuals. There was no evidence for association in the total sample or in subsets stratified by age or APOE epsilon4 status. These results suggest that this polymorphism is not a strong genetic risk factor for either early- or late-onset forms of the disorder. However, they do not exclude the possibility that an AD susceptibility allele is located elsewhere in A2M or a nearby gene. PMID- 10477120 TI - Genetic association between alpha-2 macroglobulin and Japanese sporadic Alzheimer's disease. AB - Alpha-2 macroglobulin (encoded by the gene A2M) is a serum pan-protease inhibitor that may be related with the pathogenesis of Alzheimer's disease (AD) because of its ability to mediate amyloid beta degradation. Recently, several groups have reported that the five-nucleotides deletion in A2M gene at the 5' splice site of exon 18 might increase risk for AD. In the present study, therefore, this mutation was studied in 69 healthy controls and 55 sporadic AD cases by polymerase chain reaction- restriction fragment length polymorphism method. The allelic frequencies with the deletion (A2M-2) are 9.4 and 6.4% in the control and AD groups, respectively. There is no significant difference in the A2M-2 frequencies between the controls and sporadic AD cases. This is the first report to study the frequencies of A2M-2 in Japanese AD cases, suggesting its no genetic association with sporadic AD. PMID- 10477121 TI - Association study of serotonin-6 receptor variant (C267T) with schizophrenia and aggressive behavior. AB - The relative abundance of serotonin type 6 receptors (5-HT6) in some limbic regions and the high affinity of some antipsychotics to 5-HT6 receptors suggest that they might be involved in the pathogenesis of schizophrenic disorders. In a population-based association study, we tested the hypothesis that the allelic variant, C267T, of the human 5-HT6 gene confers susceptibility to schizophrenic disorders and associated aggressive behavior. We genotyped 5-HT6 receptors in 186 patients with schizophrenic disorders and 163 controls. The results demonstrated no significant difference in genotype or allele frequencies between patients with or without aggressive behaviors. However, genotype distribution was significantly different between schizophrenic patients and control subjects. This suggests that the 5-HT6 gene may play a role in the pathogenesis of schizophrenic disorders. PMID- 10477122 TI - Selective inhibition of rat mesangial cell proliferation by a synthetic peptide derived from the sequence of the C2 region of PKCbeta. AB - RACK (receptor for activated C-kinase) is a protein that binds and translocates protein kinase C (PKC) to the appropriate cellular organelles. The binding of RACK has been mapped to C2 region of PKC. A number of peptides from the C2 region of PKCbeta have been shown to inhibit the translocation and activation of PKCbeta. This investigation was undertaken to study the role of PKCbeta in rat mesangial cell proliferation mediated by a number of mitogens. Exposure of rat mesangial cells to thrombin, endothelin, epidermal growth factor, and phorbol 12,13-dibutyrate resulted in increased [3H]thymidine incorporation. Pretreatment of mesangial cells with Ro 32-0432 (selective PKC inhibitor) inhibited the proliferation mediated by all the above mitogens, suggesting that these mitogens mediated proliferation through PKC. Experiments were performed to further evaluate the involvement of PKCbeta in this process by using the peptide derived from the C-2 region of PKCbeta as a tool. The data suggest that although the peptide (P) alone had no effect on basal- or mitogen-mediated proliferation, the peptide in the presence of a carrier peptide (PC) inhibited proliferation mediated by endothelin. In the same experiment, proliferation mediated by epidermal growth factor, thrombin and phorbol dibutyrate was unaffected, suggesting that in rat mesangial cells, endothelin mediated proliferation through the activation of PKCbeta. PMID- 10477123 TI - Antimicrobial peptides from the Brazilian frog Phyllomedusa distincta. AB - Different peptides were purified by chromatographic procedures from the skin secretory glands of the frog Phyllomedusa distincta. These are the first peptides reported from this frog species. Their primary structure was determined by a combination of automated Edman degradation and mass spectrometry. Peptide Q2 contains 25 amino acid residues, peptide Q1 and L have 28 each, peptide M contains 31, and peptide K has 33 amino acid residues. They all showed potent antimicrobial activity against Gram-negative and Gram-positive bacteria, presenting minimal inhibitory concentrations from 0.6 to 40 microM, when tested against Enterococcus faecalis, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Peptides K, L, and Q1 were chemically synthesized and shown to be active. PMID- 10477124 TI - Locustatachykinin isoforms in the locust: distribution and quantification in the central nervous system and action on the oviduct muscle. AB - The presence of locustatachykinin (LomTK)-like immunoreactivity is demonstrated in the central nervous system (CNS) of Locusta migratoria with the use of a polyclonal antiserum raised against LomTK1. By developing a radioimmunoassay with the same antiserum, we have demonstrated picomolar amounts of LomTK-like material in the tissues of the central nervous system. In contrast, only femptomolar amounts of LomTK-like material are associated with the oviduct tissue. The relative amounts of the different LomTK isoforms in the brain and the abdominal ganglionic chain were examined by separating the native peptides on high performance liquid chromatography and comparing their retention times to synthetic LomTK standards. The amounts of the different isoforms of LomTK differed between and within the two regions of the central nervous system. However, the ratios of the different isoform amounts were similar between the two regions. The myostimulatory activities of LomTKs 1 to 4 were characterized by using the locust oviduct bioassay. LomTKs 1, 2, and 3 appeared to be more efficacious than LomTK4. PMID- 10477125 TI - Structure, distribution, and biological activity of novel members of the allatostatin family in the crayfish Orconectes limosus. AB - In the central and peripheral nervous system of the crayfish, Orconectes limosus, neuropeptides immunoreactive to an antiserum against allatostatin I (= Dipstatin 7) of the cockroach Diploptera punctata have been detected by immunocytochemistry and a sensitive enzyme immunoassay. Abundant immunoreactivity occurs throughout the central nervous system in distinct interneurons and neurosecretory cells. The latter have terminals in well-known neurohemal organs, such as the sinus gland, the pericardial organs, and the perineural sheath of the ventral nerve cord. Nervous tissue extracts were separated by reverse-phase high-performance liquid chromatography and fractions were monitored in the enzyme immunoassay. Three of several immunopositive fractions have been purified and identified by mass spectroscopy and microsequencing as AGPYAFGL-NH2, SAGPYAFGL-NH2, and PRVYGFGL NH2. The first peptide is identical to carcinustatin 8 previously identified in the crab Carcinus maenas. The others are novel and are designated orcostatin I and orcostatin II, respectively. All three peptides exert dramatic inhibitory effects on contractions of the crayfish hindgut. Carcinustatin 8 also inhibits induced contractions of the cockroach hindgut. Furthermore, this peptide reduces the cycle frequency of the pyloric rhythms generated by the stomatogastric nervous system of two decapod species in vitro. These crayfish allatostatin-like peptides are the first native crustacean peptides with demonstrated inhibitory actions on hindgut muscles and the pyloric rhythm of the stomatogastric ganglion. PMID- 10477126 TI - Neuroendocrine peptides (insulin, pancreatic polypeptide, neuropeptide Y, galanin, somatostatin, substance P, and neuropeptide gamma) from the desert tortoise, Gopherus agassizii. AB - The traditional view that Testudines (tortoises and turtles) should be regarded as the surviving clade of the anapsid reptiles rather than classified with the diapsid reptiles (snakes, lizards, and crocodiles) has recently been challenged. Neuropeptide Y, neuropeptide gamma, and somatostatin-14 were isolated from an extract of the brain, substance P and galanin from an extract of the intestine, and insulin and pancreatic polypeptide from an extract of the pancreas of the desert tortoise, Gopherus agassizii. Despite that crocodilians did not appear until the late Triassic, the amino acid sequences of the tortoise peptides resemble those of the American alligator quite closely. The primary structures of neuropeptide Y, somatostatin-14, and neuropeptide gamma are the same in tortoise and alligator. The primary structures of substance P, insulin, galanin, and pancreatic polypeptide in the two species differ by 1, 3, 5, and 8 amino acid residues, respectively. Although fewer neurohormonal peptides from squamates (lizards and snakes) have been characterized, the primary structures of neuropeptide gamma, insulin, and pancreatic polypeptide from the Burmese python and the desert tortoise differ by 3, 8, and 18 residues, respectively. The data suggest, therefore, a closer phylogenetic relationship between Testudines and Crocodilians than that derived from 'classical' analyses based on morphological criteria and the fossil record. PMID- 10477128 TI - Prior pregastric food stimulation and gastrin-releasing peptide1-27 (GRP) synergize to inhibit sham feeding. AB - The hypothesis that prior pregastric food stimulation is sufficient to reveal an inhibitory effect of gastrin-releasing peptide1-27 (GRP) on sham feeding was tested in 11 male rats equipped with chronic gastric cannulas. Rats were sham fed a high-carbohydrate solution during a 45-min test session, after 17-h food deprivation. GRP (16 or 32 microg/kg) or saline was injected intraperitoneally either at the onset or 5 or 15 min after the onset of sham feeding. This allowed for a 0-, 5-, or 15-min period of pregastric food stimulation before GRP or saline injections. Sham intake was recorded every 5 min, and behavior was observed every minute. GRP inhibited sham feeding when it was administered after 5 or 15 min of prior pregastric food stimulation, but not when it was administered at test onset. A nonsignificant increase in resting behavior and decrease in feeding behavior were associated with the decrease in sham feeding. No anomalous behaviors were noted. We conclude that a synergy between GRP and prior pregastric, presumably oral, food stimulation is sufficient to inhibit sham feeding. PMID- 10477127 TI - Bombesin induces a reduction of somatostatin inhibition of adenylyl cyclase activity, Gi function, and somatostatin receptors in rat exocrine pancreas. AB - To analyze the effect of bombesin on the somatostatin (SS) mechanism of action in the exocrine pancreas, male Wistar rats (250-270 g) were injected intraperitoneally with bombesin (10 microg/kg) three times daily at 8-h intervals for 7 or 14 days. Bombesin attenuated the ability of SS to inhibit forskolin stimulated adenylyl cyclase activity in pancreatic acinar membranes. However, it did not decrease the ability of forskolin to stimulate the adenylyl cyclase catalytic subunit. The ability of 5'-guanylylimidodiphosphate [Gpp(NH)p] (a nonhydrolyzable GTP analog) to inhibit forskolin-stimulated adenylyl cyclase activity was diminished in pancreatic acinar cell membranes from bombesin-treated rats. Bombesin administration did not affect the ADP-ribosylation of a 41-kDa G protein catalyzed by pertussis toxin. The maximal SS binding capacity of pancreatic acinar membranes from bombesin-treated rats was decreased when compared with controls at the two time periods studied. The bombesin/gastrin releasing peptide antagonist [D-Tpi6,Leu13psi(CH2NH)Leu14]bombesin (6-14) (RC 3095) (10 microg/kg i.p.), injected three times daily at 8-h intervals for 7 or 14 days, had a similar effect to that of bombesin on the SS mechanism of action. The combined administration of bombesin and its antagonist RC-3095 had a greater effect on the SS receptor-effector system than when administered separately. The present study indicates that the pancreatic SS receptor-effector system may be regulated by bombesin in vivo. PMID- 10477129 TI - Gastrin-releasing peptide suppresses independent but not intraoral intake. AB - Independent and intraoral intake tests have been used to separate the effects of various substances on the appetitive and consummatory phases of ingestive behavior. This study compared the ability of gastrin-releasing peptide1-27 (GRP) to suppress intraoral intake of nutrient solutions versus independent intake of the same solutions from a bottle. In a series of experiments, adult male Sprague Dawley rats implanted with anterior sublingual chronic intraoral catheters were injected intraperitoneally with saline control or 28 microg/kg GRP before 20-min intraoral and 20-min one-bottle intake tests of a sucrose (0.1 M) and milk solution (1.2 kcal/ml). GRP potently reduced independent intake of both sucrose and milk from a bottle but had no significant effect on intraoral intake of either solution. From these results, we conclude that GRP affects appetitive related aspects of the feeding process to reduce food intake. PMID- 10477130 TI - Effects of estradiol, progesterone, and norethisterone on regional concentrations of galanin in the rat brain. AB - Concentrations of immunoreactive galanin were compared in eight gross brain regions of ovariectomized female rats treated with either estradiol, estradiol + progesterone, estradiol + norethisterone, or placebo. Higher concentrations with estradiol treatment compared with placebo were found in the pituitary (357%), frontal cortex (162%), occipital cortex (174%), hippocampus (170%), and median eminence (202%). A more profound difference with addition of progesterone or norethisterone was seen in the pituitary (529% and 467%, respectively). Sex steroids, particularly estradiol, modulate galanin concentrations not only in reproductive, but also in nonreproductive, brain regions. PMID- 10477131 TI - In vivo effects of 2-substituted [Mpa1Sar7Arg8]-oxytocin antagonists on postpartum rat. AB - A set of oxytocin antagonists consisting of [Mpa1Sar7Arg8]-oxytocin substituted by various conformationally restricted or bulky D amino acids at position 2 were synthetized and biologically tested. In in vivo pharmacological investigations, the effects of these peptides were examined on the spontaneous motor activity of postpartum rat. Three of the newly prepared peptides proved at least as effective in inhibiting uterine contractions as clinically investigated atosiban. PMID- 10477132 TI - Intravenous lysine vasopressin lowers body temperature in normal and febrile pigs. AB - Vasopressin has been implicated as a centrally acting endogenous antipyretic. However, in several species, including the pig, plasma vasopressin concentrations increase during the early stages of fever. This experiment investigated the effects of intravenous lysine vasopressin on core temperature in normal and febrile swine. Lysine vasopressin (20 microg/pig) stimulated cortisol release and induced a 60-min hypothermic episode in normal animals, although a 10-fold lower dose was without effect. The peptide also delayed the pyretic response to bacterial endotoxin (20 microg intravenously). It is speculated that the hypothermic action of circulating vasopressin may involve nitric oxide. PMID- 10477133 TI - Modulatory effects of PLG and its peptidomimetics on haloperidol-induced catalepsy in rats. AB - A behavioral model of dopaminergic function in the rat was used to examine the anticataleptic effects of L-prolyl-L-leucyl-glycinamide (PLG) and peptidomimetic analogs of PLG. Administration of 1 mg/kg PLG intraperitoneally significantly attenuated haloperidol (1 mg/kg)-induced catalepsy (as measured by the standard horizontal bar test), whereas doses of 0.1 and 10 mg/kg PLG did not. Eight synthetic PLG peptidomimetics (Calpha, alpha-dialkylated glycyl residues with lactam bridge constraint [1-4] and without [5-8]) were tested in the same manner (at a dose of 1 microg/kg) and categorized according to their activity, i.e. very active (5), moderately active (2, 3, 4, and 6), and inactive (1, 7, and 8). The catalepsy-reversal action of the diethylglycine-substituted peptidomimetic 5 was examined further and found to exhibit a U-shaped dose-response effect with an optimal dose of 1 microg/kg. The similarity between the effects of PLG and the synthetic peptidomimetics suggests a common mechanism of action. Finally, the synthetic peptidomimetics examined here, particularly peptidomimetic 5, were more effective than PLG in attenuating haloperidol-induced catalepsy. PMID- 10477134 TI - Induction of adrenomedullin by hypoxia in cultured human coronary artery endothelial cells. AB - To explore the role of adrenomedullin (ADM) in pathophysiology of ischemic heart disease, we investigated the effects of hypoxia on the production and secretion of ADM in cultured human coronary artery endothelial cells. Treatment with hypoxia (5% CO2/94% N2/1% O2) for 6 and 12 h increased expression levels of ADM mRNA 2.2-fold and fivefold compared with the normoxia control, respectively. The levels of immunoreactive ADM in the media were increased by 12-h hypoxia about fivefold compared with the control (39.0+/-1.1 fmol/10(5) cells per 12 h under hypoxia and 7.9+/-0.4 fmol/10(5) cells per 12 h under normoxia; P<0.01, n = 4, mean +/- SEM). Reverse-phase high-performance liquid chromatography of the extracts of culture media under normoxia and hypoxia showed one major peak eluting in the position of human ADM standard. The production and secretion of ADM were increased in cultured human coronary artery endothelial cells under hypoxia. ADM may therefore play an important pathophysiological role in ischemic heart disease. PMID- 10477135 TI - Peptides and the origin of life. AB - Considering the state-of-the-art views of the geochemical conditions of the primitive earth, it seems most likely that peptides were produced ahead of all other oligomer precursors of biomolecules. Among all the reactions proposed so far for the formation of peptides under primordial earth conditions, the salt induced peptide formation reaction in connection with adsorption processes on clay minerals would appear to be the simplest and most universal mechanism known to date. The properties of this reaction greatly favor the formation of biologically relevant peptides within a wide variation of environmental conditions such as temperature, pH, and the presence of inorganic compounds. The reaction-inherent preferences of certain peptide linkages make the argument of 'statistical impossibility' of the evolutionary formation of the 'right' peptides and proteins rather insignificant. Indeed, the fact that these sequences are reflected in the preferential sequences of membrane proteins of archaebacteria and prokaryonta distinctly indicates the relevance of this reaction for chemical peptide evolution. On the basis of these results and the recent findings of self replicating peptides, some ideas have been developed as to the first steps leading to life on earth. PMID- 10477136 TI - Immunolocalization of the four prostaglandin E2 receptor proteins EP1, EP2, EP3, and EP4 in human kidney. AB - Four prostaglandin E2 receptor subtypes designated EP1, EP2, EP3, and EP4 have been shown to mediate a variety of effects of prostaglandin E2 (PGE2) on glomerular hemodynamics, tubular salt and water reabsorption, and on blood vessels in the human kidney. Despite the important role of renal PGE2, the localization of PGE2 receptor proteins in the human kidney is unknown. The present study used antipeptide antibodies to the EP1 to EP4 receptor proteins for immunolocalization in human kidney tissue. Immunoblot studies using these antibodies demonstrated distinct bands in membrane fraction from human kidney. By means of immunohistochemistry, expression of the human EP1 receptor subtype protein in renal tissue was detected mainly in connecting segments, cortical and medullary collecting ducts, and in the media of arteries and afferent and efferent arterioles. The human EP2 receptor subtype protein was detectable only in the media of arteries and arterioles. The human EP3 receptor subtype protein was strongly expressed in glomeruli, Tamm-Horsfall negative late distal convoluted tubules, connecting segments, cortical and medullary collecting ducts, as well as in the media and the endothelial cells of arteries and arterioles. Staining of the human EP4 receptor subtype protein was observed in glomeruli and in the media of arteries. However, no signal of either receptor subtype was detected in the thick ascending limb, the macula densa, or in adjacent juxtaglomerular cells. These results support the concept that PGE2 modulates specific functions in different anatomical structures of the human kidney. PMID- 10477137 TI - Immunolocalization of protein kinase C isoenzymes alpha, beta1 and betaII in rat kidney. AB - Protein kinase C (PKC) significantly contributes to the control of renal function, but little is known about the renal function or localization of PKC isoenzymes. Therefore, the localization of PKC isoenzymes alpha, betaI, and betaII was studied in rat kidney. Immunoblot analysis identified immunoreactive bands corresponding to PKC a, betaI, and betaII in total cell extracts of both renal cortex and medulla. Immunohistochemistry using confocal laser scanning microscopy revealed immunostaining for PKC alpha within the glomerulus including podocytes and mesangial cells. PKC betaI was detected in mesangial cells, whereas anti-PKC betaII labeled neither podocytes nor mesangial cells. PKC betaII, however, was detected in cells within the mesangial area, which expressed MHC II, a marker for antigen-presenting cells. None of the three isoforms was detected in glomerular endothelial cells. A prominent immunostaining with anti-PKC alpha and betaI was localized to the brush border of S2 and S3 segments of proximal tubule, whereas S 1 segments were not stained. Along the loop of Henle, both PKC a and PKC betaI were found in the luminal membrane of cortical and medullary thick ascending limb. In addition, anti-PKC betaI labeled the luminal membrane of thin limbs. In the cortical collecting duct (CCD), immunofluorescence for PKC alpha was observed at the apical membrane of both peanut agglutinin (PNA)-negative cells and part of PNA-positive cells, whereas in the medullary collecting duct (MCD), PKC a was detected at the basolateral membrane. In comparison, PKC betaI was localized at the luminal membrane of PNA-positive cells only in CCD and at the luminal membrane of MCD. Unlike PKC a or betaI, there was (1) no detectable immunostaining with anti-PKC betaII in the proximal tubule, the loop of Henle, or the CCD and (2) a distinct staining for PKC betaII of interstitial cells in cortex and medulla (including MHC II-positive dendritic cells). Furthermore, PKC betaII was detected in the luminal membrane of MCD. In summary, a distinct and differential expression pattern for PKC alpha, betaI, and betaII was shown in rat kidney, which may contribute to a better understanding of the specific role of these isoenzymes in the control of renal function. PMID- 10477138 TI - Ser-322 is a critical site for PKC regulation of the MDCK cell taurine transporter (pNCT). AB - Previous studies have shown that the Madin-Darby canine kidney cell taurine transporter (pNCT) is downregulated by protein kinase C (PKC) activation. In this study, it is hypothesized that the highly conserved serine-322 (Ser-322) located in the fourth intracellular segment (S4) may play an important role in the function of taurine transporter, which is modulated by PKC phosphorylation. It is demonstrated that Ser-322 is the critical site of PKC phosphorylation, as determined by site-directed mutagenesis. When Ser-322 of pNCT was changed to alanine (S322A) and this mutant was evaluated in an oocyte expression system, taurine transport activity increased threefold compared with control (wild-type pNCT). Activation of PKC by the active phorbol ester 12-myristate 13-acetate did not influence taurine transport by mutant S322A. Kinetic analysis showed that the mutation of Ser-322 essentially changed the Vmax, rather than the Km, of the transporter. Mutation of all other PKC consensus sites did not affect transporter activity when expressed in the oocyte system. Western blot analysis showed that expression of taurine transporter protein was similar in oocytes injected with either wild-type or mutant pNCT cRNA, indicating that the enhanced taurine transport activity by mutant S322A was not caused by a greater amount of transporter expressed in the oocyte. Furthermore, this study demonstrated that the taurine transporter was phosphorylated after PKC activation, and this effect was not observed in mutant S322A. In conclusion, Ser-322 is critical in PKC regulation of taurine transporter activity. The steady-state taurine transporter activity is tightly controlled by endogenous PKC phosphorylation of Ser-322, which is located in the fourth intracellular segment of the taurine transporter. PMID- 10477139 TI - Short-term pravastatin mediates growth inhibition and apoptosis, independently of Ras, via the signaling proteins p27Kip1 and P13 kinase. AB - Growth factor-stimulated DNA synthesis in a variety of cell lines has been shown to be decreased after overnight (or longer) treatment with the 3-hydroxy-3 methylglutaryl CoA reductase inhibitors, the statins. Although this anti mitogenic effect had been presumed to be the result of the impairment of Ras lipidation, a stable modification (T1/2 approximately 20 h), this study provides new data demonstrating that brief (approximately 1 h) pretreatment of rat vascular smooth muscle cells with 100 microM pravastatin before platelet-derived growth factor-BB (PDGF-BB) stimulation results in attenuation of DNA synthesis through a Ras-independent mechanism. PDGF-BB-stimulated PDGF-beta receptor tyrosine phosphorylation, Ras activity, and mitogen-activated protein/extracellular signal-regulated kinase activity are unaffected by from 10 min to 1 h of pravastatin incubation, while Raf activity is markedly increased after 1 h of pravastatin. Phosphatidylinositol-3 kinase activity and phosphorylation of its downstream effector Akt are decreased after 1 h pravastatin incubation. Rho is stabilized by pravastatin, and ADP-ribosylation of Rho by C3 exoenzyme decreases PDGF-stimulated phosphatidylinositol-3 kinase activity, mimicking the effect of pravastatin on this signaling protein. Levels of the cyclin-dependent kinase inhibitor p27Kip1 are increased when cells were preincubated with pravastatin for 1 h and then exposed to PDGF, and apoptosis is induced by pravastatin incubation times as short as 1 to 4 h. Thus, short-term, high-dose pravastatin inhibits vascular smooth muscle cell growth and induces apoptosis independently of Ras, likely by means of the drug's effect on p27Kip1, mediated by Rho and/or phosphatidylinositol-3 kinase. This work demonstrates for the first time that the statins may be therapeutically useful when applied for short periods of time such that potential toxicity of long-term statin use (such as chronic Ras inhibition) may be avoided, suggesting future therapeutic directions for statin research. PMID- 10477140 TI - High glucose stimulates proliferation and collagen type I synthesis in renal cortical fibroblasts: mediation by autocrine activation of TGF-beta. AB - Renal tubular epithelial cells and interstitial fibroblasts are active participants in tubulointerstitial fibrosis, the best correlate of decreased glomerular filtration in diabetic nephropathy. It was reported previously that high ambient glucose stimulates transforming growth factor-beta (TGF-beta) mRNA and bioactivity, promotes cellular hypertrophy, and increases collagen synthesis in proximal tubular cells. This study evaluates the effects of high glucose and TGF-beta on the behavior of murine renal cortical fibroblasts (TFB) in culture. High glucose (450 mg/dl) significantly increased [3H]-thymidine incorporation (by 60 to 80% after 24 to 72 h) and cell number, without significantly increasing cell death when compared with normal glucose (100 mg/dl). There also was a transient increase in the mRNA of the c-myc and egr-1 early-response genes. Exogenous TGF-beta1 was promitogenic rather than antiproliferative in contrast to other renal cell types. Northern blot analysis demonstrated constitutive expression of TGF-beta1, -beta2, and -beta3 transcripts. Exposure to high glucose increased all three TGF-beta isoforms in a time-dependent manner. High glucose as well as exogenous TGF-beta1 also increased [3H]-proline incorporation, alpha2(I) collagen mRNA, and type I collagen protein (measured by immunoassay). Treatment with a neutralizing pan-selective monoclonal anti-TGF-beta antibody markedly attenuated the stimulation by high ambient glucose of thymidine incorporation, TGF-beta1 mRNA, and type I collagen mRNA and protein levels. It is concluded that high ambient glucose and exogenous TGF-beta1 share similar actions on renal fibroblasts. Moreover, the stimulation of cell proliferation and collagen type I synthesis in these cells by high ambient glucose are mediated by activation of an autocrine TGF-beta system. PMID- 10477141 TI - Focal segmental glomerulosclerosis: prognostic implications of the cellular lesion. AB - The cellular lesion (CELL), seen in some patients with primary focal segmental glomerulosclerosis (FSGS), comprises proliferation, hypertrophy, and pathologic changes in the cells overlying the glomerular scar. The prognosis of the cellular lesion was retrospectively studied in 100 patients with FSGS (43 had FSGS-CELL and 57 had FSGS without the cellular lesion (classic segmental scar [CS]). Patients with the FSGS-CELL lesion were more often black and severely proteinuric and developed more end-stage renal disease (ESRD). Nephrotic patients with FSGS CELL (n = 39) were more proteinuric at presentation than patients with FSGS-CS (n = 36). ESRD developed more frequently in patients with the FSGS-CELL (17 of 39, 44% versus 5 of 36, 14%, P = 0.005), and patients with extensive FSGS-CELL (> or = 20% glomeruli) were mainly black (94%), severely nephrotic (67%, >10 g/d), and had a poor response to treatment (23% remission). In nephrotic patients, initial serum creatinine, interstitial expansion > or = 20%, and CELL independently predicted ESRD. However, the rates of remission in treated nephrotic patients with FSGS-CELL and FSGS-CS were the same (9 of 17, 53% versus 17 of 39, 52%), and patients in both groups who achieved a remission had a 5-yr survival of 100%. Steroid treatment was the only variable that predicted remission. Patients with the FSGS-CELL have an increased prevalence of ESRD, but the improved prognosis associated with remission is so significant that a therapeutic trial is warranted in all nephrotic FSGS patients, regardless of the presence of the cellular lesion. PMID- 10477142 TI - Transduction of renal cells in vitro and in vivo by adeno-associated virus gene therapy vectors. AB - There has been an increasing interest recently in the possibility of treating renal diseases using gene therapy. The ability to pursue gene therapy for renal diseases has been limited by the availability of an adequate system for gene delivery to the kidney. Adeno-associated virus (AAV) is a defective virus of the parvovirus family that has a number of properties attractive for renal gene delivery: recombinant AAV contains no viral genes; expression of genes delivered by these vectors does not activate cell-mediated immunity; the virus is able to transduce nondividing as well as dividing cells; and both wild-type and recombinant AAV integrate into the host chromosome resulting in long-term gene expression. Studies were performed to determine whether AAV can deliver reporter genes to kidney cells in vitro and in vivo. These studies show that AAV can deliver reporter genes with approximately equal efficiency to human mesangial, proximal tubule, thick ascending limb, collecting tubule, and renal cell carcinoma cells in primary culture. Immortalized mouse mesangial cells are transduced at a much greater efficiency. Transduction can be enhanced by pharmaceutical agents up to sevenfold in primary cells (transducing up to 20% of primary cells per well) and as much as 400-fold in immortalized mesangial cells. AAV delivered in vivo by intraparenchymal injection results in at least 3 mo of reporter gene expression in tubular epithelial, but not glomerular or vascular, cells at the injection site. These data indicate that AAV can deliver genes to renal cells both in vitro and in vivo resulting in prolonged gene expression, and thus AAV can be a useful tool for renal gene delivery. PMID- 10477143 TI - The angiotensin-converting enzyme genotype and microalbuminuria in autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (AD-PKD) has a variable clinical course. Clinical parameters associated with a worse prognosis are hypertension and proteinuria or microalbuminuria (MA). Because chronic stimulation of the renin-angiotensin system is likely to be present in ADPKD patients, the effect of the angiotensin-converting enzyme insertion/deletion (ACE I/D) genotype on the variability of these clinical parameters was examined in untreated ADPKD patients. Proteinuria and MA were determined in 24-h urine collections. BP measurements were performed with an ambulatory monitor, over 24 h. With analysis of covariance, the ACE genotype was found to be significantly associated with MA, corrected for age, gender, GFR, mean arterial pressure, body surface area, and urinary Na+ excretion (P < 0.05). The patients homozygous for the deletion (DD) had the highest rate of MA (P < 0.05) compared to the patients homozygous for the insertion (II). There was no relationship between the ACE genotype and BP or renal function. A significant positive correlation was found between MA and mean arterial pressure (r = 0.31, P < 0.05), whereas a significant negative correlation was found between MA and renal function (r = -0.28, P < 0.05). In conclusion, in ADPKD patients, MA is partly determined by the ACE I/D polymorphism. Because MA is associated with an enhanced progression toward renal failure, the ACE genotype could help in identifying patients at risk for a worse prognosis. PMID- 10477144 TI - Worldwide ethnic distribution of the G protein beta3 subunit 825T allele and its association with obesity in Caucasian, Chinese, and Black African individuals. AB - Recently, it was demonstrated that one allele (825T) of the gene encoding the G protein beta3 subunit (GNB3) is associated with hypertension in Germans. This study investigates a possible association with obesity in young male Germans, Chinese, and black South Africans with low, intermediate, and high 825T allele frequencies, respectively. In each of these three distinct cohorts, the 825T allele frequency was increased significantly in overweight (body mass index [BMI] > or =25 kg/m2) and obese individuals (BMI >27 kg/m2) compared to those with normal weight. The 825T allele frequencies in these three BMI groups were, respectively, 29.5, 39.3, and 47.7% in Germans, 46.8, 53.9, and 58.6% in Chinese, and 83.1, 87.7, and 90.9% in South Africans. In each of these three distinct groups, the 825T allele was significantly associated with obesity with odds ratios between 2 and 3. More urban than rural black Africans were overweight despite similar 825T allele frequencies in both populations, which underscores the role of both genetic and environmental factors. BP values in young male whites increased significantly with increasing BMI values but were independent of the C825T polymorphism, suggesting that hypertension associated with the 825T allele could be a consequence of obesity. Genotyping of 5254 individuals from 55 native population samples from Africa, the Americas, Europe, Asia, Australia, and New Guinea demonstrated highest 825T allele frequencies in black Africans (82%) and intermediate values in east Asians (47%). It is anticipated that high frequencies of the 825T allele in Africans and Asians may contribute to an obesity and hypertension epidemic if Westernization of lifestyles continues. PMID- 10477145 TI - Regulation of renal laminin in mice with type II diabetes. AB - This study examines the regulation of renal laminin in the db/db mouse, a model of type II diabetes characterized by extensive remodeling of extracellular matrix. Immunohistochemistry demonstrated an increase in the contents of laminin chains including beta1 chain in the mesangium and tubular basement membranes at 1, 2, 3, and 4 mo of diabetes. Immunofluorescence with an antibody against the recently discovered laminin alpha5 chain showed that in the normal mouse, the protein had a restricted distribution to the glomerular and tubular basement membranes with scant expression in the mesangium of older mice. In the diabetic mouse, the laminin alpha5 chain content of the glomerular and tubular basement membranes was increased, with marked expression in the mesangium. Northern analysis revealed a significant decrease in the renal cortical contents of alpha5, beta1, and gamma1 chain mRNA in the diabetic mice compared to control, at each of the time points. In situ hybridization showed decreased abundance of alpha5 transcripts in the glomeruli of diabetic mice compared to nondiabetic controls. Analysis of mRNA changes by Northern and in situ hybridization studies demonstrated that the reduction in laminin transcripts involved both glomerular and tubular elements. These observations demonstrate that laminin accumulation in the db/db mice with type II diabetes is due to nontranscriptional mechanisms. Because previous investigations in rodents with type I diabetes have shown that the increase in renal laminin content was associated with a corresponding increment in laminin chain transcript levels, it appears that the mechanisms underlying augmentation in renal matrix laminin content may be distinct in the two types of diabetes. PMID- 10477146 TI - Role of caspases in hypoxia-induced necrosis of rat renal proximal tubules. AB - The role of the caspases, a newly discovered group of cysteine proteases, was investigated in a model of hypoxia-induced necrotic injury of rat renal proximal tubules. An assay for caspases in freshly isolated rat proximal tubules was developed. There was a 40% increase in tubular caspase activity after 15 min of hypoxia in association with increased cell membrane damage as indicated by a threefold increase in lactate dehydrogenase release. The specific caspase inhibitor Z-Asp-2,6-dichlorobenzoyloxymethylketone (Z-D-DCB) attenuated the increase in caspase activity during 15 min of hypoxia and markedly decreased lactate dehydrogenase release in a dose-dependent manner. In the proximal tubules, Z-D-DCB also inhibited the hypoxia-induced increase in calpain activity, another cysteine protease. In contrast, when Z-D-DCB was added to purified calpain in vitro, there was no inhibition of calpain activity. The calpain inhibitor (2)-3-(4-iodophenyl)-2-mercapto-2-propenoic acid (PD150606) also inhibited the hypoxia-induced increase in caspase activity in proximal tubules, but did not inhibit the activity of purified caspase 1 in vitro. In these experiments, caspase activity was detected with the fluorescence substrate Ac-Tyr Val-Ala-Asp-7-amido-4-methyl coumarin (Ac-YVAD-AMC), which is preferentially cleaved by caspase 1. However, minimal caspase activity was detected with the fluorescence substrate Ac-Asp-Glu-Val-Asp-7-amido-4-methyl coumarin (Ac-DEVD AMC), which is cleaved by caspases 2, 3, and 7. The present study in proximal tubules demonstrates that (1) caspase inhibition protects against necrotic injury by inhibition of hypoxia-induced caspase activity; and (2) caspase 1 may be the caspase involved. Thus, although the role of caspases in apoptotic cell death is well established, this study provides new evidence that caspases contribute to necrotic cell death as well. PMID- 10477147 TI - Renal expression of aquaporins in liver cirrhosis induced by chronic common bile duct ligation in rats. AB - Semiquantitative immunoblotting was used to investigate the expression levels of the four major renal aquaporins, the Na-K-2Cl cotransporter of the thick ascending limb, the type 3 Na-H exchanger, and the Na-K-ATPase in kidneys from rats with cirrhosis secondary to common bile duct ligation (CBDL). These rats had significant water retention and hyponatremia. In contrast to models of cirrhosis induced by carbon tetrachloride, aquaporin-2 expression in CBDL-induced cirrhosis was decreased. Thus, these results show that in the setting of extracellular fluid volume expansion, excessive water retention with hyponatremia can occur in the absence of increases in aquaporin-2 abundance. In addition, the expression levels of the two basolateral collecting duct aquaporins (aquaporin-3 and -4) were decreased in CBDL rats relative to sham-operated control rats. Similarly, the Na-K-2Cl cotransporter of the thick ascending limb and the type 3 Na-H exchanger showed decreases in expression. In contrast, the expression levels of aquaporin-1 and the all subunit of the Na-K-ATPase were not decreased. Thus, dysregulation of multiple water channels and ion transporters may play a role in water balance abnormalities associated with CBDL-induced cirrhosis in rats. PMID- 10477148 TI - Clinical presentation and follow-up of 30 patients with congenital nephrogenic diabetes insipidus. AB - Congenital nephrogenic diabetes insipidus is characterized by insensitivity of the distal nephron to arginine vasopressin. Clinical knowledge of this disease is based largely on case reports. For this study, data were collected on clinical presentation and during long-term follow-up of 30 male patients with congenital nephrogenic diabetes insipidus. The majority of patients (87%) were diagnosed within the first 2.5 yr of life. Main symptoms at clinical presentation were vomiting and anorexia, failure to thrive, fever, and constipation. Three older patients were diagnosed as a result of events not directly related to the disease. Except for a possibly milder phenotype in patients with a G185C mutation, no clear relationship between clinical and genetic data could be found. Most patients were on hydrochlorothiazide-amiloride treatment without significant side effects. Two patients suffered from severe hydronephrosis with a small rupture of the urinary tract after a minor trauma, and two patients experienced episodes of acute urine retention. Height SD scores for age remained below the 50th percentile in the majority of patients, whereas weight for height SD scores showed a catch-up after several years of underweight. PMID- 10477150 TI - Heart rate variability during chronic hemodialysis and after renal transplantation: studies in patients without and with systemic amyloidosis. AB - The present study was undertaken to compare heart rate variability (HRV) values in patients on maintenance hemodialysis with no evidence of ischemic or hypertensive heart diseases to those of age- and gender-matched healthy individuals and those of patients after renal transplantation. To assess the effects of a common confounding factor, HRV values were also determined in patients with systemic amyloidosis, in chronic hemodialysis, and after successful renal transplantation. Spectral analyses of RR intervals from continuous electrocardiogram recordings were performed to quantify ultra low frequency, very low frequency, low frequency, and high frequency powers. HRV determinations were all significantly reduced in uremic patients undergoing hemodialysis compared with the healthy control subjects, especially in those with systemic amyloidosis. Renal transplantation normalized HRV in most patients; HRV, however, remained reduced in isolated amyloidosis patients with cardiac or adrenal involvement. HRV circadian day/night differences were preserved in hemodialysis patients and after renal transplantation in those without amyloidosis but not in those with amyloidosis. These data suggest that reduced HRV in chronic hemodialysis patients may precede other manifestations of cardiovascular disease. In uremic patients with amyloidosis, a more severe form of autonomic failure may occur. Successful transplantation corrects HRV abnormalities in most patients, suggesting that the autonomic dysfunction of uremia is caused by humoral factors reversed by the normalization of the renal function. PMID- 10477149 TI - Mycophenolate mofetil for maintenance therapy of Wegener's granulomatosis and microscopic polyangiitis: a pilot study in 11 patients with renal involvement. AB - Successful maintenance therapy with mycophenolate mofetil (MMF) 2 g/d and low dose oral corticosteroids (OCS) over a period of 15 mo was given to patients with Wegener's granulomatosis (WG) (n = 9) and microscopic polyangiitis (MPA) (n = 2). All patients had severe generalized disease with pauci-immune necrotizing glomerulonephritis and received standard induction therapy with oral cyclophosphamide and OCS for a mean of 14 wk until remission was achieved. Of 11 patients, only one WG patient relapsed in the 14th month of maintenance therapy. Maintenance therapy with MMF was able to further reduce grumbling disease activity as measured by the Birmingham vasculitis activity score (BVAS2) and proteinuria that were still present by the end of induction therapy. OCS could be reduced to a median daily dose of 5 mg and discontinued in three patients. Possible drug-related adverse effects were transient and included abdominal pain, respiratory infection, diarrhea, leukopenia, and a cytomegalovirus-colitis in one patient that was successfully treated with ganciclovir. It is concluded that MMF in combination with low-dose OCS is well tolerated and effective for maintenance therapy of WG and MPA. Long-term treatment with MMF in these diseases is attractive because of its low toxicity. MMF will have to be studied further and compared with cyclophosphamide or azathioprine maintenance therapy in randomized trials. PMID- 10477151 TI - Increased erythrocyte phosphatidylserine exposure in chronic renal failure. AB - The appearance of phosphatidylserine, an aminophospholipid normally confined to the inner monolayer, at the outer leaflet of red cell membrane may have several pathophysiologic implications. This study examines erythrocyte phosphatidylserine exposure in chronic renal failure (CRF) patients on conservative treatment or on dialysis, to assess possible alterations to phospholipid asymmetry in a condition associated with a state of deranged red cell function. A significant increase in phosphatidylserine-expressing erythrocytes was found in undialyzed patients with CRF (2.32%) and patients on hemodialysis (3.06%) and on peritoneal dialysis (2.14%) compared with control subjects (0.68%). In undialyzed CRF patients, a strong correlation (r = 0.903) was found between the percentage of phosphatidylserine-expressing red cells and the serum creatinine concentration. The increased exposure of phosphatidylserine in uremic erythrocytes may be due to inhibition of phosphatidylserine transport from the outer to the inner leaflet of plasma membrane and may promote an increased erythrophagocytosis. In reconstitution experiments, normal erythrocytes showed an increase in phosphatidylserine-expressing cells when incubated in uremic plasma (3.2% after 2 h versus 1.1% at beginning of incubation), whereas phosphatidylserine-positive uremic erythrocytes decreased when resuspended in normal plasma (2.03% after 2 h and 1.65% after 8 h versus 2.9% at beginning of incubation). Preliminary characterization of the putative uremic compound(s) indicates a molecular weight between 10,000 and 20,000, as well as heat instability. These findings show an impairment of erythrocyte membrane phospholipid asymmetry in CRF patients, regardless of the dialysis treatment. Such abnormality seems related to the uremic state and could contribute to the red cell pathology present in CRF. PMID- 10477152 TI - Activation of 5-lipoxygenase and related cell membrane lipoperoxidation in hemodialysis patients. AB - Lipid peroxidation was shown at the membrane level in peripheral blood cells of patients hemodialyzed on cuprophan dialyzers, and was mainly attributable to the generation of conjugated hydroperoxides in the lipid bilayer. The oxidative index (i.e., the A234/205 ratio) of membrane lipids was 3.2-fold higher in hemodialysis patients than in healthy control subjects, and also the level of leukotriene B4 was significantly increased (up to 1.7-fold over control). Both membrane peroxidation and release of leukotriene B4 were linked to upregulation of 5 lipoxygenase activity (up to 2.4-fold over control) and expression at the protein level (up to 1.9-fold). Vitamin E, the most important lipophilic antioxidant, prevented both membrane peroxidation and release of leukotriene B4 by inhibiting 5-lipoxygenase activity without affecting enzyme expression. Similar results were observed in patients hemodialyzed on polymethylmetacrylate membranes, but in this case the activation of 5-lipoxygenase was less pronounced. The use of a purified 5-lipoxygenase demonstrated that vitamin E was a reversible inhibitor of enzyme activity (IC50 = 35 +/- 4 microM), further characterized as noncompetitive (Ki = 30 +/- 3 microM). Taken together, the results reported here shed some light on the mechanism responsible for the oxidative damage in hemodialysis. Moreover, the beneficial effect of vitamin E described here may have relevance for the therapy of patients with kidney disease. PMID- 10477153 TI - In vitro and in vivo immunomodulatory effects of RDP1258, a novel synthetic peptide. AB - Peptides derived from certain regions of human class I MHC molecules are known to have immunomodulatory effects. In particular, amino acid residues 75-84 of the HLA-B7 and HLA-B2702 molecules have demonstrated allele nonspecific immunosuppression in several animal transplant models. There is evidence that these effects are mediated by binding to intracellular heat shock proteins, including heme oxygenase-1. A new derivative of these peptides, RDP1258, was developed using a novel computer-assisted rational design technique. In vitro, RDP1258 peptide inhibited rat heme oxygenase activity in a dose-dependent manner. Similar to observations made with other in vitro heme oxygenase inhibitors, in vivo administration of RDP1258 peptide to naive rats resulted in upregulation of splenic heme oxygenase activity. The effects of the peptide on alloimmune responses were then tested. Addition of RDP1258 to rat and human mixed leukocyte reactions inhibited proliferation in a dose-dependent manner. In a rat renal transplantation model, peptide therapy combined with a sub-therapeutic dose of cyclosporin A significantly prolonged allograft survival. These data provide further evidence that modulation of the heat shock protein heme oxygenase by rationally designed peptides affects immune effector functions and may allow the development of novel immunomodulatory strategies in organ transplantation. PMID- 10477154 TI - Apoptosis of tubular epithelial cells in donor kidney biopsies predicts early renal allograft function. AB - Acute renal failure (ARF) is a serious complication in the early postoperative period after kidney transplantation. In an effort to identify subjects at risk, several donor-, recipient-, and procedure-related factors have been studied. Because no morphologic parameter predictive of delayed graft function has been identified to date, this study was conducted to determine whether the number of apoptotic cells in donor biopsies obtained before engraftment is predictive of the development of posttransplant ARF. Donor biopsies of patients with "biopsy proven" acute tubular damage but no signs of rejection (n = 23) showed significantly higher counts of apoptotic tubular epithelial cells when compared to patients with immediate transplant function (n = 44) or early rejection (n = 22). In all groups, a significantly higher percentage of apoptotic cells was found in the distal compared to the proximal tubule. The expression of bcl-2 and proliferating cell nuclear antigen was not different among the groups. Late allograft function was not affected by early ARF as serum creatinine values were similar in all three groups after 6 mo. These data suggest that the number of apoptotic renal tubular epithelial cells in donor biopsies before engraftment is predictive of the early postoperative course in patients undergoing kidney transplantation. PMID- 10477155 TI - Effect of protein A immunoadsorption in nephrotic syndrome of various etiologies. AB - Protein A immunoadsorption (IA) has proved effective in reducing proteinuria in patients with nephrotic syndrome after recurrence of focal and segmental glomerulosclerosis (FSGS) in kidney transplants. The effect of IA in nephrotic syndrome of other etiologies remains unknown. Nine patients with nephrotic syndrome secondary to membranous nephropathy (four cases), diabetes mellitus (one case), IgA nephropathy (two cases), and amyloidosis (two cases) had three to five IA of 2.5 plasma volumes over 4 to 8 d. Patients received no concomitant immunosuppressive treatment, and antihypertensive drugs were left unchanged. Proteinuria decreased from 12.64 +/- 5.49 to 3.35 +/- 2.2 g/24 h (mean +/- SD) in all patients after three to five IA. Hematocrit decreased from 37.32 to 32.64% (12.5% hemodilution) and serum albumin from 25.43 to 18.6 g/L (26.4% decrease). Proteinuria returned to baseline levels within 1 mo, as described in recurrent FSGS following transplantation. When serum albumin balance was controlled by albumin infusion after IA in two patients, comparable decreases in proteinuria were observed. Therefore, IA is effective in producing short-term reduction of proteinuria in nephrotic syndromes related not only to FSGS but also to membranous and IgA nephropathies, diabetes mellitus, and amyloidosis, which suggests that IA removes a nonspecific circulating hemodynamic-altering or permeability-increasing factor. PMID- 10477156 TI - How they begin and how they end: classic and new theories for the development and deterioration of congenital anomalies of the kidney and urinary tract, CAKUT. AB - CAKUT are problems that often require surgical intervention or, in the worst case, lead to renal failure and the need for dialysis and/or renal transplantation. It is believed that these anomalies share a common genetic cause and to date there has been no good animal model with which to study these abnormalities. Although the abnormal interaction between the ureteral bud and metanephric blastema leads to renal hypodysplasia, vesicoureteral reflux, and ectopic ureters to name a few, the genetic and biochemical modulation of urinary tract development is not understood. Studies using the mouse strain mutant for angiotensin type 2 (AT2) receptors have given new insight into this mystery. The animals show defective apoptosis of undifferentiated mesenchymal cells in the area surrounding the developing kidney and urinary tract. This abnormal apoptosis may well interfere with the normal interaction between the ureteral bud and metanephric blastema resulting in CAKUT. This abnormal interaction would theoretically lead to preexisting intrinsic abnormalities of the kidney, which are programmed and take effect early in embryonic development. In the worst cases, the renal abnormalities would lead to progressive deterioration of renal function. Undoubtedly, there are more genes and biochemical modulators involved in this process other than the RAS and AT2 receptors. Our current animal model gives new and unique possibilities with which to study development of the kidney and urinary tract and ultimately seek ways of preventing an often debilitating disease process. PMID- 10477157 TI - An indistinct balance: the safety and efficacy of parenteral iron therapy. AB - Recombinant epoetin therapy and correction of the chronic anemia of renal failure have greatly reduced the number of red cell transfusions and hence the propensity to iron overload. The majority of HD patients require intravenous iron therapy to achieve the hematocrit levels that correspond to improved outcome measures. Although the short-term benefits of intravenous iron have been clearly defined, the long-term risks of intravenous iron are less well-defined. Iron overload before the availability of epoetin constituted a serious problem; our review of the literature does not decisively conclude that these patients had more serious bacterial infections or increased mortality when compared with their non-iron overloaded counterparts, unless chronic transfusion-related hepatic disease was superimposed. Specifically, no data unequivocally confirm that iron overload from parenteral iron contributes to all-cause patient morbidity or mortality. Furthermore, therapy that maintains intravenous iron optimal iron stores and replaces iron losses associated with the dialytic procedure does not engender iron overload in the carefully monitored patient. Optimized anemia therapy in ESRD requires individualized and specific application of epoetin and iron for each patient, and significant cost savings can result from such a strategy. Prospective studies are clearly necessary to define those parameters that reflect adequacy of iron storage in renal failure patients. We should develop alternative means of iron delivery and develop monitors that accurately discriminate between patients who will respond to additional iron therapy and those who will not. Whether ferritin should be supplanted by another parameter and whether iron itself poses an increased risk to those patients it has so beneficially served are issues that must be resolved. Until these answers are known, the importance of carefully crafted iron therapy cannot be overstated. PMID- 10477158 TI - 1998 Presidential Address. PMID- 10477159 TI - Disease of the month. The Churg Strauss Syndrome. AB - The Churg Strauss Syndrome is an eosinophil-associated small vessel vasculitis. Although its pathogenesis may be distinctive and the association with severe late onset asthma typical, the clinical features during the vasculitic phase widely overlap with those of the other forms of necrotizing vasculitis, and no single clinical or histologic feature is pathognomic of the condition. Renal involvement is common, although usually mild, and even when severe it tends to respond well to treatment. The prognosis for both patient and renal survival with adequate treatment is in general good. The optimal treatment strategy, however, is uncertain, and may differ from that for the other vasculitides. In particular, in contrast to Wegener's granulomatosis, the need for routine cyclophosphamide treatment is unconfirmed and requires further study. PMID- 10477160 TI - Active transport of sodium as the source of electric current in the short circuited isolated frog skin. Reprinted from Acta. Physiol. Scand. 23: 110-127, 1951. PMID- 10477161 TI - ATP-based tumor chemosensitivity testing: assisting new agent development. AB - Chemotherapy of cancer based on cytotoxic agents has proved successful in the treatment of many cancers. The number of agents available to the oncologist has grown steadily and drug combinations are in widespread use. The perceived success of these combinations makes the introduction of new agents difficult. For any new agent, multiple phase II and III trials are likely to be needed. Since phase II/III trials usually only address single issues, the cost of introducing a new agent is substantial. Multiple studies are required with different tumor types to define the activity profile of a new drug, followed by adjusted combinations to define the role of the new drug in conjunction with older ones. Recent advances in the understanding of cancer at a molecular level are already leading to new agent design. The next problem is how to introduce and use these agents. One possible approach is to trial the drugs with tumor cells ex vivo, using a chemosensitivity assay such as the ATP-based chemosensitivity assay which is designed to mimic the situation within the tumor accurately enough to examine issues of dose response, sequence and timing in many different tumors. The avoidance of cell lines ensures relevance and the sensitivity of some of these methods allows large numbers of mechanistically logical permutations to be tested with material from small numbers of patients. The results may be used to choose the most effective combinations for clinical testing in a limited number of subsequent phase II/III trials, saving money and time, while permitting new agents to be introduced faster. PMID- 10477162 TI - Heterogeneity of chemosensitivity of metastatic cutaneous melanoma. AB - Advanced melanoma has a poor prognosis and chemotherapy provides little benefit for most patients. This may be related to heterogeneity of chemosensitivity as well as frequent constitutive resistance to individual cytotoxic drugs. We have therefore examined the heterogeneity of chemosensitivity in metastatic cutaneous melanoma specimens using an ex vivo ATP-based chemosensitivity assay (ATP-TCA). Melanoma deposits (n=55) in skin or lymph node were tested using the ATP-TCA, performed in three separate laboratories. Analysis of the data collected (based on an arbitrary sensitivity index < 300) shows considerable heterogeneity of chemosensitivity. The most active single cytotoxic agents in the assay were identified as cisplatin, treosulfan, paclitaxel, vinblastine, gemcitabine and mitoxantrone. There was also a limited direct inhibition of melanoma cell growth by interferon-alpha2b, although this agent is known to have a number of indirect biological antitumor effects. Exposure of tumor cells to combinations of drugs at the concentrations tested as single agents showed the most active combinations to be treosulfan+gemcitabine, cisplatin+paclitaxel and vinblastine+paclitaxel. There was considerable heterogeneity of chemosensitivity: some tumors responded well to one agent or combination, while others showed no response to this and instead responded to one of the alternatives tested. Occasional highly resistant tumors showed no response to any of the single agents or combinations tested. The degree of heterogeneity observed suggests that the ATP-TCA could be used to select patients who might benefit from specific chemotherapeutic agents alone or in combination. This provides the rationale for future randomized controlled trials of ATP-TCA-directed chemotherapy versus physician's choice to determine whether assay-directed chemotherapy can improve patient response and survival. PMID- 10477163 TI - Selective cell kill of the combination of gemcitabine and cisplatin in multilayered postconfluent tumor cell cultures. AB - Both gemcitabine (2',2'-difluorodeoxycytidine, dFdC) and cisplatin (cis-diammine dichloroplatinum) have significant anticancer activity against ovarian, head and neck, and non-small cell lung cancer (NSCLC). dFdC can be incorporated into DNA and RNA, and inhibit DNA repair, while cisplatin can form Pt-DNA adducts. We previously observed schedule-dependent synergism of the combination of dFdC and cisplatin in monolayer cell cultures. We now evaluated whether the combination would also enable selective cell kill in multilayered postconfluent cell cultures, since each compound showed variable activity in multilayered cells. The combination was tested in multilayered cultures from cell lines with a different histological origin: the human head and neck squamous cell carcinoma cell line UMSCC-22B (22B), the human NSCLC cell line H322, and ADDP, a cisplatin-resistant variant of the human ovarian cancer cell line A2780. Sensitivity of the multilayered cells was dependent on exposure duration and sequence of the drug combinations, which were added in a constant molar ratio (dFdC:cisplatin 1:100), with a total exposure time of 96 h. The type of interaction was related to the degree of resistance of the cell lines to either dFdC or cisplatin. Thus, the very sensitive 22B cells only showed an additive effect when cells were preincubated for 24 h with dFdC prior to exposure to the combination. In contrast, in the resistant ADDP and H322 cells, synergism was the most common profile (three out of four schedules tested). This is of special relevance when we take into account that these drugs only show cytostatic effects when administered alone, whereas the combination produced cytotoxic cell killing. In conclusion, combining dFdC with cisplatin can be at least additive, but synergistic in multilayered postconfluent cells resistant to dFdC and cisplatin. PMID- 10477164 TI - Phase II study of oral trofosfamide as palliative therapy in pretreated patients with metastatic soft-tissue sarcoma. AB - This phase II study investigated the activity of continuously administered oral trofosfamide in chemotherapy-pretreated patients with metastatic soft-tissue sarcoma (STS). Trosfosfamide is an oxazaphosphorine with ifosfamide as the predominant metabolite. Eighteen patients with a median age of 60 years were treated with trofosfamide given as continuous oral treatment. Starting dose was 300 mg/day for 7 days and subsequently 150 mg/day. All patients had previously received at least one chemotherapy regimen including doxorubicin and ifosfamide. Three patients achieved partial responses (18%) and nine a disease stabilization (53%) for an overall response rate of 18% (95% CI: 0.5-35%). Median progression free interval was 4 months (0-17 months) and median overall survival was 10 months (4-39+) months. Toxicity was generally mild. Only one WHO grade III nausea, but no other non-hematologic WHO grade III/IV toxicity occurred. Leukopenia WHO grade III/IV was observed in four patients (22%). No thrombocytopenia <50,000/microl and no neutropenic infection was seen. Continuously administered oral trofosfamide is a well-tolerated palliative treatment in anthracycline/oxazaphosphorin-pretreated patients with advanced STS achieving responses and/or disease stabilization in up to 70% of patients. PMID- 10477165 TI - Cisplatin-resistant derivatives of murine L1210 leukemia cells are not susceptible to growth-inhibiting and apoptosis-inducing actions of transforming growth factor-beta1. AB - Murine L1210 leukemia cells possessing an increased resistance to cisplatin were found to be refractory to transforming growth factor (TGF)-beta1-induced growth inhibition, while the parental L1210 cells were strongly inhibited by this cytokine. Growth inhibition was estimated on the basis of [3H]thymidine incorporation, cell counting and colony-forming assay. Cisplatin-resistant L1210 cells were also shown to be much more resistant than the parental cells to both cisplatin- and TGF-beta1-induced apoptosis. These results suggest the existence of cross-resistance to cisplatin and TGF-beta1 in the studied leukemia cells. PMID- 10477166 TI - A double-blind randomized study comparing intramuscular (i.m.) granisetron with i.m. granisetron plus dexamethasone in the prevention of delayed emesis induced by cisplatin. The Italian Multicenter Study Group. AB - Granisetron has been shown to exert a beneficial therapeutic effect in the prophylaxis and treatment of acute nausea and vomiting due to chemotherapy. However, limited data regarding its efficacy in the prevention and treatment of delayed emesis are available. A total of 532 patients entered this multicenter double-blind study, aimed at comparing the efficacy and safety of intramuscular (i.m.) granisetron with that of i.m. granisetron plus dexamethasone. Complete response and total control were evaluated for 3 days following the first 24 h after cisplatin administration in two groups of patients: 262 treated with granisetron 3 mg i.m. daily (plus placebo), and 265 with granisetron at the same dose plus dexamethasone 8 and 4 mg twice daily. The rate of complete response was 58.0% in the granisetron group and 78.9% in the granisetron plus dexamethasone group over days 1-3 (p<0.01). Similarly, over the same period total control was 44.7% with granisetron alone and 65.3% with granisetron plus dexamethasone (p<0.01). Local and systemic tolerability of the i.m. therapy with granisetron were satisfactory. In conclusion, granisetron plus dexamethasone showed good protection against delayed emesis due to emetogenic chemotherapy. PMID- 10477167 TI - DNA methylation of retinoic acid receptor beta in breast cancer and possible therapeutic role of 5-aza-2'-deoxycytidine. AB - The retinoic acid receptor beta (RARbeta), a putative tumor suppressor gene, has been reported to be poorly expressed in breast cancer. In this report using the methylation-specific PCR reaction we observed DNA methylation in the promoter region of RARbeta in several primary breast tumors. DNA sequence analysis showed that the positions of 5-methylcytosine in the RARbeta promoter region was almost identical to that reported previously by our laboratory for human DLD-1 colon carcinoma cells (Anti-Cancer Drugs 1998; 9: 743). Several other cancer-related genes have been also reported to be silenced by DNA methylation, including the p16 tumor suppressor gene, E-cadherin, an invasion suppressor gene and the estrogen receptor gene in breast cancer cell lines. Since breast cancer cells have several potential target genes for the DNA methylation inhibitor, 5-aza-2' deoxycytidine (5-Aza-CdR), we investigated the in vitro antineoplastic activity of this analog on the human breast cancer cell line MDA-MB-231. We report that 5 Aza-CdR is a potent growth inhibitor and a potent cytotoxic agent against the breast carcinoma cells. These results suggest that 5-Aza-CdR may be an interesting agent to investigate in patients with breast cancer resistant to conventional chemotherapy. PMID- 10477168 TI - Microtubule-disrupting effects of gallium chloride in vitro. AB - Gallium chloride (GaCl3), an antitumor agent with antagonistic action on iron, magnesium and calcium, was tested for its ability to alter the polymerization of purified tubulin (2.2 mg/ml) in a cell-free system in vitro. GaCl3 (250 microM) does not mimic the effect of 10 microM paclitaxel and, therefore, is not a microtubule (MT)-stabilizing agent that can promote tubulin polymerization in the absence of glycerol and block MT disassembly. In contrast, GaCl3 mimics the effect of 1 microM vincristine (VCR) and inhibits glycerol-induced tubulin polymerization in a concentration-dependent manner (IC50: 125 microM), indicating that GaCl3 is a MT de-stabilizing agent that prevents MT assembly. However, 150 microM GaCl3 must be used to match or surpass the inhibitions of tubulin polymerization caused by 0.25 microM of known MT de-stabilizing agents, such as colchicine (CLC), nocodazole, podophyllotoxin, tubulozole-C and VCR. The inhibitory effect of 250 microM GaCl3 persists in the presence of up to 9 mM MgCl2, suggesting that the exogenous Mg2+ cations absolutely required for the binding of GTP to tubulin and MT assembly cannot overcome the antitubulin action of Ga3+ ions of a higher valence. The binding of [3H]vinblastine (VBL) to tubulin (0.5 mg/ml) is inhibited by unlabeled VBL but enhanced by concentrations of GaCl3 > 200 microM. However, increasing concentrations of GaCl3 mimic the ability of cold CLC to reduce the amount of [3H]CLC bound to tubulin, suggesting that GaCl3 may interact with the CLC binding site to inhibit tubulin polymerization. The binding of [3H]GTP to tubulin is decreased by unlabeled GTP but markedly enhanced by GaCl3, especially when concentrations of this metal salt of 32 microM or higher are added to the reaction mixture before rather than after the radiolabeled nucleotide. These data suggest that changes in protein conformation following GaCl3 binding might increase the interactions of tubulin with nucleotides and Vinca alkaloids. After a 24 h delay, the viability of GaCl3 treated L1210 leukemic cells is reduced in a concentration-dependent manner at days 2 (IC50: 175 microM), 3 (IC50: 35 microM) and 4 (IC50: 16 microM). Since GaCl3 (100-625 microM) increases the percentage of mitotic cells at 2-4 days, it might arrest tumor cell progression in M phase, but its antimitotic activity is much weaker than that of 0.25 microM VCR. Because the concentrations of GaCl3 that inhibit tubulin polymerization also increase the mitotic index and decrease the viability of L1210 cells in vitro, the antitubulin and antimitotic effects of GaCl3 might contribute, at least in part, to its antitumor activity. PMID- 10477169 TI - Tricyclic pyrone analogs: a new synthetic class of bifunctional anticancer drugs that inhibit nucleoside transport, microtubule assembly, the viability of leukemic cells in vitro and the growth of solid tumors in vivo. AB - Tricyclic pyrones (TPs) may represent a novel synthetic class of microtubule (MT) de-stabilizing anticancer drugs previously shown by us to inhibit macromolecule synthesis, tubulin polymerization, and the proliferation of leukemic and mammary tumor cells in vitro. A linear skeleton with a N-containing aromatic ring attached at C3 of the top A-ring, a central pyran B-ring and a six-membered bottom C-ring with no alkylation at C7 are required for the antitumor activities of the lead compounds, a 3-pyridyl benzopyran (code name H10) and its somewhat weaker 2-pyridyl regioisomer (code name H19). Increasing concentrations of H10 do not alter the binding of [3H]vinblastine and [3H]GTP to tubulin but mimic the ability of unlabeled colchicine (CLC) to reduce the amount of [3H]CLC bound to tubulin, suggesting that TPs may interact with the CLC binding site to inhibit tubulin polymerization. Exogenous Mg2+ cations absolutely required for the binding of GTP to tubulin and MT assembly cannot overcome the antitubulin action of H10. H10 reduces the viability of L1210 cells in vitro (IC50: 0.5 microM) but its antitumor activity may be related to its ability to inhibit tubulin polymerization and rapidly increase the mitotic index rather than to induce DNA cleavage and apoptosis. The anticancer potential of TPs in vivo is demonstrated by the fact that i.p. injections of the water-soluble H10-HCl decrease the growth of solid tumors in mice inoculated s.c. with Lewis lung carcinoma. A critical finding is that the antimitotic H10 is a bifunctional anticancer drug, which also blocks the cellular transport of nucleosides (IC50: 6 microM) to inhibit DNA synthesis. Since few CLC site-binding antimitotic agents are active in solid tumor models in vivo, the ability of these new MT destabilizing TPs to totally block nucleoside transport might be valuable in polychemotherapy to arrest tumor cells at several phases of their cycle, potentiate the action of antimetabolites and sensitize multidrug-resistant tumor cells. PMID- 10477170 TI - Variable intrinsic sensitivity of human tumor cell lines to raltitrexed (Tomudex) and folylpolyglutamate synthetase activity. AB - The cytotoxic effects of Tomudex (TX) were investigated on a panel of 15 human tumor cell lines expressing a spontaneous sensitivity to the tested agent. We determined the basal cellular amount of relevant cellular factors potentially related to the cytotoxic efficacy of or resistance to TX. We selected thymidylate synthase (TS) as the target for TX, basal reduced folates (RF), because RF may compete with TX for a common site on the TS molecule. We also tested folylpolyglutamate synthetase (FPGS) because this is the enzyme which transforms the drug into its active polyglutamated form. Results were as follows. There was a wide inter-cell line variability in IC50 values for TX and there were marked differences between cell lines for all tested biochemical parameters. No link was observed between basal cellular TS activity and TX cytotoxic efficacy. There was an inverse relationship between reduced folate cellular content and TX IC50 values; this relationship did not, however, reach statistical significance. The only significant relationship was found between basal cellular FPGS activity and TX IC50r = -0.56, p = 0.03. Tumor cells with a relatively high FPGS activity were more sensitive to TX cytotoxic effects and vice versa. Along with previous results which showed that acquired resistance to TX is accompanied by a decrease in FPGS activity, the present data are strongly indicative of a prominent role played by FPGS activity in the intrinsic sensitivity to TX. Means to up-regulate FPGS activity with pharmacological or tumor-specific genetic approaches are recommended so as to optimize TX antitumor activity. PMID- 10477171 TI - Altered tyrosine phosphorylation of ERK1 MAP kinase and other macrophage molecules caused by Leishmania amastigotes. AB - The involvement of tyrosine phosphorylation during macrophage infection with Leishmania amazonensis amastigotes was investigated. PTK antagonists such as genistein, herbimycin A, geldanamycin and tyrphostin 25 had no significant effect on adhesion to, or entry into, murine peritoneal macrophages, but increased parasite intracellular survival. LPS-induced tyrosine phosphorylation of target host proteins assessed by immunoprecipitation and Western blot was impaired or reversed by living amastigotes soon after 60 min-infection. Such reversion was not due to parasite-secreted molecules but was contact-dependent, as assessed by cytochalasin D treatment of macrophage monolayers prior to infection. Paraformaldehyde-fixed or sodium vanadate-treated amastigotes exerted no significant effect on overall macrophage tyrosine phosphorylation. Immunoprecipitation of proteins employing 4G10 anti-phosphotyrosine antibody followed by Western blotting revealed that tyrosine phosphorylation of 120, 85, 60, 44 and 35 kDa proteins was selectively reversed by amastigote infection. Inhibition, measured by densitometry was from about 66-100% of uninfected cells. None of these proteins was immunoprecipitated from amastigote-infected macrophage lysates but all of them except for p85 were recovered after treatment of parasites with 100 microM sodium orthovanadate prior to infection, a treatment that inhibits Leishmania amastigote protein ecto-phosphatase. The 44 kDa protein was identified as ERK1 MAP kinase (MAPK) by Western blot. Amastigote infection also decreased tyrosine phosphorylation induced by zymosan particles. Vanadate treatment of amastigotes prior to infection significantly decreased parasite intracellular survival. The action of a putative leishmanial ecto-protein phosphatase (PPase) is suggested. PMID- 10477172 TI - Intragenic recombinants of Plasmodium falciparum identified by in situ polymerase chain reaction. AB - We report an in situ PCR technique for visualising amplified DNA of blood forms of Plasmodium falciparum on microscope slides by fluorescence microscopy. The method is used to assess the changes in frequency of different alleles of the MSP1 gene in cultures of the progeny of a cross. We show that parasites with a recombinant form of this protein possess an initial growth advantage before declining in numbers over the long-term. PMID- 10477173 TI - Molecular and biochemical characterization of a protein kinase B from Trypanosoma cruzi. AB - A Trypanosoma cruzi gene, PKB, coding for a putative protein kinase was cloned and sequenced. Analysis of the sequence showed that the encoded protein (called PKB) corresponds to a relatively novel subgroup of Ser/Thr protein kinases denominated protein kinases B (PKB), related to A and C protein kinases (RAC), or protein kinases of the transforming retrovirus AKT8 (Akt) in which the catalytic domains show similarity to corresponding domains of protein kinases A and protein kinases C. Unlike mammalian enzymes belonging to the same subgroup, PKB did not have a pleckstrin (PH)-homologous domain. PKB was expressed in Escherichia coli and the recombinant protein was found to be a Thr-specific protein kinase that required Mn2+ for activity and used ATP as phosphate donor (Km = 1.8 microM). Classical protein kinase A and protein kinase C modulators and inhibitors were found to have only marginal or no effect on PKB activity. Antisera raised against the recombinant protein recognized PKB in Western blotting analysis of cell extracts as a membrane bound protein. Evidence was obtained suggesting the presence of a Cys-linked acyl anchor. Northern and Western blotting analysis showed that PKB was constitutively expressed in the lag, exponential and stationary phases of T. cruzi epimastigote growth, as well as in the amastigote and metacyclic trypomastigote stages of differentiation. This is the first description of the existence of a protein kinase B in trypanosomatid protozoa. PMID- 10477174 TI - Differential regulation of ESAG transcripts in Trypanosoma brucei. AB - In Trypanosoma brucei, several genes termed ESAGs for expression site-associated genes are contained within the polycistronic transcription units of the VSG gene, and their transcription is coordinately regulated during the parasite life-cycle. Since the VSG mRNA is characterized by a drastic destabilization under conditions where translation is inhibited, we examined if this post-transcriptional control also applies to the ESAG mRNAs. While the ESAG 7/6 mRNA behaved like the VSG mRNA, the ESAG 8 and ESAG 3 mRNAs did not. We ascribe this differential behaviour to the residual transcription that still occurs only in the ESAG 7/6 region of the VSG unit under conditions where this unit is down-regulated. PMID- 10477175 TI - A new potent antigen from Echinococcus granulosus associated with muscles and tegument. AB - An immunoscreening of a cDNA library derived from the adult stage of the parasitic platyhelminth Echinococcus granulosus has been carried out with sera from infected dogs. The EgA31 clone encodes a fibrous protein which shares some sequence elements with paramyosins. The corresponding gene is present as a single copy in the genome. As revealed by an antibody obtained against a fusion protein produced in bacteria, the polypeptide has a molecular weight of 66 kDa. This polypeptide is present at all developmental stages studied and is a potent antigen during an infection by the adult stage in the dog and during cyst growth in human patients. By immunohistology, it was shown that it is present in the tegument and subtegumental parenchyma of the adult with a main location in the region of the suckers where it rapidly accumulates at the time of the head evagination. It is also present in the germinal layer of the cyst and on the protoscolex. PMID- 10477177 TI - Regulation of Entamoeba invadens encystation and gene expression with galactose and N-acetylglucosamine. AB - Encystation of Entamoeba invadens parasites is prevented by the presence of free galactose or N-acetylglucosamine in the encystation medium. Galactose prevents the formation of amoeba cellular aggregates which develop during the early phase of encystation, suggesting the presence of functional cell surface galactose binding molecules, whereas N-acetylglucosamine allows aggregation to occur and prevents cyst formation at a later point. While studying sugar inhibition of amoeba encystation, it was found that high efficiency encystation required the inclusion in encystation medium of precise amounts of polyvalent galactose terminated molecules, and these molecules could be supplied by serum or by defined glycoconjugates, including mucin. Addition of free galactose to encystation medium prevented the accumulation of three transcripts which are normally upregulated during encystation, and N-acetylglucosamine prevented accumulation of one of the transcripts. These results suggest the presence of distinct sugar-sensitive pathways that regulate differentiation of the amoeba trophozoite into infectious cysts. PMID- 10477176 TI - Expression of Trypanosoma cruzi surface antigen FL-160 is controlled by elements in the 3' untranslated, the 3' intergenic, and the coding regions. AB - The FL-160 surface antigen gene family of T. cruzi consists of hundreds of members of 160 kDa glycoproteins expressed in trypomastigotes, but not in epimastigotes. Steady-state levels of FL-160 mRNA were 80 to 100-fold higher in trypomastigotes than in epimastigotes, yet transcription rates were equivalent between the lifecycle stages. Luciferase reporter constructs demonstrated that the 3' untranslated region (UTR) and intergenic region (IR) following the coding sequence of FL-160 was sufficient to generate 8-fold higher luciferase expression in trypomastigotes compared with epimastigotes. Transfection of 3' UTR/IR deletion constructs revealed cis-acting elements which conferred a trypomastigote specific expression pattern similar to that of FL-160. Parasites treated with translation and transcription inhibitors, cyclohexamide and Actinomycin D, respectively, displayed a stage-specific pattern of FL-160 mRNA degradation. Epimastigotes, but not trypomastigotes, treated with the inhibitors accumulated a 1.4 Kb FL-160 cleavage product. The cleavage site mapped to a 31 base poly-purine tract in the FL-160 coding region. The first 526 aa of FL-160, containing the 31 base poly-purine tract and several smaller tracts, were fused to green fluorescent protein (GFP) and expressed from the T. cruzi tubulin locus. Stable transformants expressed 4-fold more FL-160:GFP fusion mRNA and 12-fold more fusion protein in the trypomastigote stage than in the epimastigote stage suggesting post-transcriptional and translational control elements. These data reveal at least two distinct control mechanisms for trypomastigote-specific expression of FL-160 surface glycoproteins, one involving the 3' UTR/IR and one involving the coding region of FL-160. PMID- 10477178 TI - Expression and characterization of elastase inhibitors from the ascarid nematodes Anisakis simplex and Ascaris suum. AB - Two elastase inhibitors, ASPI-1 and ASPI-2, from the parasitic nematode Anisakis simplex, have been isolated and characterized. Because these inhibitors are similar in size (60 amino acids in length) and primary sequence (52 and 47% identical) to the Ascaris suum chymotrypsin/elastase inhibitor-1 (AsC/E-1), we suggest that these Anisakis elastase inhibitors belong to the same unique class of canonical inhibitors formed by the family of Ascaris inhibitors (Huang K, Strynadka NCJ, Bernard VD, Peanasky RJ, James MG. Structure 1994;2:679-689). To compare ASPI-1 with AsC/E-1, we expressed both inhibitors in Pichia pastoris and found that: (1) the association constant of rASPI-1 with porcine pancreatic elastase (PPE) is similar to native inhibitor (Ka = 4.5 x 10(9) and 6.4 x 10(9) M(-1), respectively); (2) rASPI-1 is a potent inhibitor of PPE and human leukocyte elastase (Ka = 1.6 x 10(9) M(-1)); and (3) it is only a very weak inhibitor of chymotrypsin (CHYM) (Ka = 1.2 x 10(6) M(-1)). In contrast to the Anisakis inhibitor, however, rAsC/E inhibitor-1 is a very strong inhibitor of both PPE (Ka = 3.5 x 10(10) M(-1)) and CHYM (Ka = 3.6 x 10(12) M(-1)). We also found that the determined reactive sites (P1-P'1) of rASPI-1 and rAsC/E-1, as recognized by PPE, are Ala 28-Met 29 and Leu 31-Met 32, respectively. These P1 P'1 residues of AsC/E-1 constitute the same reactive site as that also recognized by CHYM (Peanasky RJ, Bentz Y, Homandberg GA, Minor ST, Babin DR. Arch Biochem Biophys 1994;232:135-142). The difference in specificities of ASPI-1 and AsC/E-1 toward their cognate serine proteases may be attributed to the P1 and P'3 residues in the inhibitors. Elastase, which recognizes both alanine and leucine, canaccommodate both ascarid inhibitors, whereas chymotrypsin, which prefers bulky, hydrophobic residues, only recognizes the Ascaris C/E inhibitor-1. PMID- 10477179 TI - Antisense oligonucleotides targeting malarial aldolase inhibit the asexual erythrocytic stages of Plasmodium falciparum. AB - A major obstacle in the global effort to control malaria is the paucity of anti malarial drugs. This is compounded by the continuing emergence and spread of resistance to old and new anti-malarial drugs in the malarial parasites. Here we describe the anti-malarial effect of phosphorothioate antisense (AS) oligodeoxynucleotides (ODNs) targeting the aldolase enzyme of Plasmodium falciparum, using the asexual blood stages of the parasite grown in vitro. The blood stages of P. falciparum depend almost entirely on the energy produced by their own glycolysis. Aldolase, the fourth enzyme of the glycolytic pathway, is highly upregulated during the malarial 48-h life cycle. We found that the mRNA of this enzyme can be inhibited, in a sequence specific manner, using AS-ODN to the splice sites on the pre-mRNA of malarial aldolase. At the enzyme level, both specific AS-ODNs for the splice sites, as well as for the translation initiation site on mature mRNA, can inhibit aldolase enzyme activity within the trophozoites of P. falciparum. Furthermore, this downregulation of the malarial aldolase results in a reduction in the production of ATP within the parasite. Finally, the treatment reduces parasitemia. In summary, AS-ODNs targeting the aldolase gene of P. falciparum can interfere with the blood-stage life cycle of this parasite in vitro by inhibiting the expression of the enzyme aldolase which results in decreased malarial glycolysis and energy production. Thus, we conclude that blockade of the expression of malarial glycolytic enzymes using specific AS-ODNs has the potential of a new anti-malarial strategy. PMID- 10477180 TI - Characterization of a SR protein from Trypanosoma brucei with homology to RNA binding cis-splicing proteins. AB - The protozoan parasite Trypanosoma brucei relies on trans-splicing to process its mRNAs. A novel nuclear serine/arginine (SR)-rich trypanosomal protein (TSR1) was characterized which contains two RNA recognition motifs. The TSR1 protein appears to be homologous to RNA-binding SR proteins of the cis-splicing machinery from higher eukaryotes. Moreover, in the yeast two-hybrid system, TSR1 is able to interact with the human splicing factors involved in the recognition of the 3' splicing site (U2AF35/U2AF65). In both procyclic and bloodstream forms of T. brucei, TSR1 was found to localize in the nucleus. In the bloodstream stage TSR1 showed the speckles pattern characteristic of SR proteins involved in cis splicing. Moreover, TSR1 was able to specifically bind the spliced leader (SL) RNA involved in trans-splicing in trypanosomes by the yeast three-hybrid system. These and other observations suggest that TSR1 may be involved in trans-splicing in T. brucei. PMID- 10477181 TI - Stage-specific expression of 14-3-3 in asexual blood-stage Plasmodium. AB - This paper reports the identification of 14-3-3 in Plasmodium. 14-3-3 is an evolutionarily conserved protein that is most noted as a mediator in signal transduction events and cell cycle regulation. The complete cDNA (approximately 2.6 kb) and gDNA (approximately 3.4 kb) of a Plasmodium knowlesi 14-3-3 (Pk14-3 3) is reported. The gene has three introns; two near the beginning and one close to the end of the coding sequence. Also reported, is the gDNA of the Plasmodium falciparum homologue (Pf14-3-3). Unlike in many other organisms, where multiple gene copies and different functional isoforms exist, Plasmodium 14-3-3 is encoded as a single-copy gene. Northern blot analyses show that the Pk14-3-3 transcript in asexual blood stages begins to be expressed in the ring-stage, predominates in young trophozoites, and thereafter declines. An antiserum produced against recombinant Pk14-3-3 reacts via immunoblot and immunoprecipitation with the approximately 30 kDa and the approximately 32 kDa Pk14-3-3 and Pf14-3-3 proteins, respectively. Protein expression in P. knowlesi closely mimics the pattern of the transcript. PMID- 10477182 TI - A Rab1 homologue with a novel isoprenylation signal provides insight into the secretory pathway of Theileria parva. AB - As a first step in developing compartment-specific markers for protein trafficking within Theileria parva, we have isolated cDNAs encoding homologues of the small GTP binding proteins Rab1 and Rab4. The T. parva homologue of Rab1 (TpRab1), a protein which regulates vesicular transport between the endoplasmic reticulum and cis golgi in other organisms, was unusual in that it contained a unique 17 amino acid C-terminal extension. The C-terminal motif sequence KCT (XCX) contrasted with the CXC or XCC motifs which act as as signals for isoprenylation by geranylgeranyl in most Rab proteins, including all known Rab1 homologues, in containing only a single cysteine. [C14]mevalonic acid lactone and [H3]geranylgeranyl pyrophosphate were specifically incorporated into recombinant TpRab1 in vitro, demonstrating that the novel motif was functional for isoprenylation. Recombinant TpRab1 bound radiolabeled GTP, and this binding was inhibited by excess unlabeled GTP and GDP and also partially by ATP. The TpRab1 gene contained four short (34-67 bp) introns with a distinct pattern of occurrence within the protein sequence as compared to the introns of other lower eukaryote Rab1 genes. Immunofluorescence microscopy using antiserum specific for the novel C-terminal peptide in combination with labelling of cells using the nucleic acid-staining dye DAPI, indicated that TpRab1 was located in the vicinity of the schizont nucleus within the infected lymphocyte. PMID- 10477183 TI - Purification and characterisation of a trypsin-like serine oligopeptidase from Trypanosoma congolense. AB - Trypanosoma brucei contain a serine oligopeptidase (OP-Tb) that is released into (and remains active in) the blood of trypanosome-infected animals. Here a similar enzyme from Trypanosoma congolense is described. This oligopeptidase, called OP Tc, was purified using three-phase partitioning, and ion-exchange and affinity chromatography. OP-Tc is inhibited by alkylating agents, by serine peptidase specific inhibitors including 3,4-dichloroisocoumarin, 4-(2 aminoethyl)benzenesulfonylfluoride and diispropylfluoro-phosphate and by other peptidase inhibitors including leupeptin, antipain and peptidyl chloromethyl ketones. Reducing agents such as dithiothreitol enhanced activity as did heparin, spermine and spermidine. The enzyme has trypsin-like specificity since it cleaved fluorogenic peptides that have basic amino acid residues (Arg or Lys) in the P1 position. Potential substrates without a basic residue in P1 were not hydrolysed. Although OP-Tc has weak arginine aminopeptidase activity, the enzyme clearly preferred substrates that had amino acids in the P2 and P3 positions. Overall, OP Tc appears to be less efficient than OP-Tb because it usually displayed lower k(cat)/Km values for the substrates tested. However, like OP-Tb, the best substrate for OP-Tc was Cbz-Arg-Arg-AMC (Km = 0.72 microM, k(cat) = 96 s(-1)). OP Tc preference for amino acids in the P2 position was (Gly,Lys,Arg) > Phe > Leu > Pro. The results also suggest that the P3-binding site has hydrophobic characteristics. OP-Tc may not be a naturally immunodominant molecule because neither IgG nor IgM anti- OP-Tc antibodies were detected in the blood of experimentally infected cattle. PMID- 10477184 TI - Characterisation of a polymorphic Tc1-like transposable element of the parasitic nematode Haemonchus contortus. AB - Hctc1, a member of the Tc1-family of transposable elements was isolated from the parasitic nematode Haemonchus contortus. Hctc1 is 1590 bp long, is flanked by 55 bp inverted repeats and carries a single open reading frame of a 340 amino acid transposase-like protein. Hctc1 is similar to Tc1 of Caenorhabditis elegans and elements Tcb1 and Tcb2 of Caenorhabditis briggsae in the inverted terminal repeats, the open reading frame, as well as the target insertion sequence. Furthermore, the copy number of Hctc1 is comparable with the Tc1 copy number in low copy strains of C. elegans. The sequence of Hctc1 is highly variable in H. contortus due to deletions, insertions and point mutations, with at least five distinct length variants of Hctc1. Most of the Hctc1 variation was within rather than between H. contortus populations. The high level of sequence variation is probably due to variation generally found for members of the Tc1-family, as well as a high background level of genetic variation of H. contortus. PMID- 10477185 TI - Analysis of Plasmodium falciparum PfEMP-1/var genes suggests that recombination rearranges constrained sequences. AB - The var genes of Plasmodium falciparum encode a family of parasite erythrocyte surface antigens, the PfEMP-1 proteins, which function as adhesion ligands for host endothelial and erythrocyte receptors. PfEMP-1 is extremely polymorphic although the extent of this variation in naturally transmitted parasite populations is unclear. We have identified 56 different sequences from the Duffy binding-like (DBL-1) domain of var genes amplified from six different P. falciparum clones isolated from patient infections in a Sudanese village in October-November 1989. These clones have been compared with 25 PfEMP-1 sequences expressed from different var gene loci by the 3D7A clone and 48 PfEMP-1 sequences from different isolates in endemic areas such as Kenya, Brazil, Gambia, Vietnam and Vanuatu to analyse diversity in clonal, local and 'global' P. falciparum populations. Evidence that certain conserved sequences recur in clones from one Sudanese village and in isolates from all over the world suggests that var gene diversity is the result of recombinational reshuffling of a subset of conserved, presumably ancestral sequences. Recurrence of particular var sequence blocks thus leads to 'overlaps' in the PfEMP-1 sequence repertoire of different P. falciparum clones. PMID- 10477186 TI - Structural studies on the polar glycoinositol phospholipids of Trypanosoma (Schizotrypanum) dionisii from bats. AB - The polar glycoinositol phospholipids (GIPLs) of a Trypanosoma species that belongs to the Schizotrypanum subgenus were purified by reversed-phase and normal phase liquid chromatography and analysed by negative-ion mode electrospray-mass spectrometry (ES-MS). The phosphatidylinositol moieties were released by nitrous acid deamination and identified as ceramide- and alkylacylglycerol-containing species. The structures of the GIPLs were determined using chemical treatments, sequential exoglycosidase digestions and positive-ion mode ES-MS-MS. All of the GIPLs were based on the same Man alpha1-2Man alpha1-2Man alpha1-6Man alpha1-4(NH2 CH2CH2-HPO3-)GlcN-PI core with single terminal Galf residue substitutions either on the terminal nonreducing Man or on the second alphaMan residue from the inositol and with either ethanolamine phosphate or 2-aminoethylphosphonate on the third alphaMan residue from the inositol. The T. (S.) dionisii GIPLs are compared with those of T. (S.) cruzi, a closely related species of the Schizotrypanum subgenus. PMID- 10477188 TI - YAC contigs and restriction maps of chromosomes 4 and 5 from the cloned line 3D7 of Plasmodium falciparum. PMID- 10477187 TI - Expression of hydrophilic surface proteins in infective stages of Leishmania donovani. PMID- 10477189 TI - A refined localization of the mitochondrial DNA primase in Crithidia fasciculata. PMID- 10477190 TI - Intensive case management of high-risk patients in a family medicine residency setting. AB - BACKGROUND: As the health care system evolves, health care delivery systems have begun to share risk for the care of patient populations. The prototype for such a system has long been the uncompensated care population in a hospital's service area. This article describes a successful case management pilot program in a family medicine residency setting. METHODS: Nineteen high-risk patients were cared for by a case management team for a period of 3 to 4 months. The case management team consisted of a medical director, 2 resident assistant medical directors, 1 registered nurse case manager, and 1 social worker. RESULTS: Case management resulted in an annualized decrease of 51 percent of inpatient days and 46 percent of charges. This resulted in an annualized savings of $166,083 in charges to the health care system. CONCLUSION: Intensive case management of the sickest of the sick results in a substantial reduction in morbidity and cost. Family medicine residency programs are ideally situated to oversee case management of this population and potentially other populations in a shared-risk environment. PMID- 10477191 TI - Simple home program for frozen shoulder to improve patients' assessment of shoulder function and health status. AB - BACKGROUND: The purpose of this investigation was to test the hypothesis that a simple home program can improve the self-assessed shoulder function and health status of a group of patients with frozen shoulders. METHODS: A case series using a one-group pretest, posttest design analyzing 41 patients from a single orthopedic practice who had a frozen shoulder were included in this study. The patients completed the Simple Shoulder Test (SST) and the Medical Outcomes Study Short-Form Health Survey (SF-36) questionnaire at the time of initial consultation, had treatment consisting of education regarding frozen shoulder and home stretching instructions, and were asked to complete the same questionnaires mailed every 6 months. Initial results were compared with previously published control values to establish level of impairment, and follow-up results were compared with the initial results to determine the extent of improvement. RESULTS: Patients initially had serious deficits in the 12 shoulder functions inventoried by the SST and were also compromised in their general health status as reflected by the SF-36 scores. At follow-up, 4 of 10 SST functions were improved (P < 0.001). The SF-36 health status scores of physical function, comfort, and physical role function were also improved (P < 0.001). CONCLUSION: These data suggest that this home program for frozen shoulder can lead to improved self-assessed shoulder function and health status in patients similar to those in the study population. PMID- 10477192 TI - Practice patterns of family physicians in practice-based research networks: a report from ASPN. Ambulatory Sentinel Pratice Network. AB - BACKGROUND: Practice-based research networks are growing and undertaking larger and more complex studies to inform the clinical practice of family physicians. We describe a study that compares clinical behaviors of physicians in the Ambulatory Sentinel Practice Network (ASPN), a large national practice-based research network, with those from the National Ambulatory Medical Care Survey (NAMCS). METHODS: A survey, replicating NAMCS, was conducted among 129 family physician members of ASPN. Nested logistic regression was used to determine which services could predict ASPN membership after adjustment for common and easily observed patient and physician characteristics. RESULTS: Of 20 specific patient services, only 4 were predictive of membership in ASPN. Of these 4, 2 were screening or diagnostic services; ASPN physicians were 1.18 times more likely to obtain a blood pressure measurement and 0.60 times as likely to order a culture for streptococcal pharyngitis. ASPN physicians were 2.30 times more likely to provide family planning counseling and 1.66 times more likely to provide smoking cessation counseling after adjusting for patient smoking status. CONCLUSIONS: We conclude that there are minimal differences in the practice patterns of family physicians participating in a large national practice-based research network and those included in the probability sample of NAMCS. Additional work is needed to examine further those characteristics of the phenomena observed in practice-based research network research that might affect generalizability of results to the larger community of practicing family physicians. PMID- 10477193 TI - Review of latex allergy. AB - BACKGROUND: Allergy to latex has become an increasing concern to many patients and health care providers. Health care providers should become aware of this problem and develop strategies for treatment and prevention. METHODS: We searched MEDLINE from 1990 to 1998 for the topics of latex allergy using the key words "latex" and "allergy." Other sources were found from back-referencing these references and from the Internet. RESULTS: Latex allergy is emerging as a clinical and occupational health problem. Understanding the clinical and immunologic features, which range from dermatitis to anaphylaxis, can provide the basis of preventive and therapeutic strategies. CONCLUSION: The clinician should be able to recognize latex allergy and educate patients about the potential hazards of latex-containing products. Providers can develop a plan for protecting patients and health care staff from latex exposure as well as treating allergic reactions when they occur. PMID- 10477194 TI - Using opioid analgesics to manage chronic noncancer pain in primary care. AB - BACKGROUND: Opioid analgesics have been considered the drugs of choice for the treatment of moderate to severe cancer and postoperative pain. Cultural attitudes and concerns about abuse have inhibited their use for a larger population of patients who have chronic noncancer pain. METHODS: Medical literature was searched from 1990 to 1998 using the key words "opioid analgesics," "opioid abuse," and "chronic pain." I have also drawn from more than 20 years of experience prescribing opioid analgesics for chronic pain. A case series analysis of 30 patients using opioid analgesics for periods ranging from 6 months to 17 years was performed. RESULTS AND CONCLUSIONS: The World Health Organization has published a "three-step ladder" guide to treatment of cancer pain graded from mild to severe; this guide can be readily applied to the treatment of pain from all sources. Opioid analgesics are the mainstay of treatment for moderate to severe pain. Chronic pain patients offer some difficult challenges for busy primary care physicians. The unique pharmacologic characteristics of opioid medications are important in outpatient management. A four-sheet office management protocol helps to focus efficiently on important clinical issues related to pain control, to monitor for opiate abuse, and to incorporate pain management more effectively in the overall primary care plan of the patient. PMID- 10477195 TI - Rational use of cholinesterase activity testing in pesticide poisoning. AB - BACKGROUND: The use of acetylcholinesterase (AChE) activity testing in pesticide poisoning often falls on family physicians when evaluating a suspected poisoning or when monitoring the health of pesticide applicators. METHODS: A review of the literature and consideration of three illustrative cases shows misunderstandings in the pathophysiology of the enzyme and in procedures for effective testing and monitoring of AChE levels. RESULTS AND CONCLUSIONS: The physiologic characteristics of acetylcholine neurotransmission are described and related to carbamate and organophosphate poisoning. Pre-exposure monitoring is described using the California plan. A 23 percent variance in AChE levels exists among normal patients. It is necessary, therefore, to establish baseline levels to overcome individual variance. The practice of measuring of AChE levels in acute poisoning is limited. In employees who have been monitored and for whom baseline AChE levels have been established, a diagnosis of poisoning can be made by comparing postexposure AChE levels with baseline levels. If there is no baseline level recorded, and if the offending chemical is in question, the clinician must base treatment on the clinical signs and symptoms. PMID- 10477196 TI - Critical appraisal of the literature. PMID- 10477197 TI - Ear cellulitis and rash in a patient with rheumatoid arthritis and diabetes. PMID- 10477198 TI - Hepatic adenoma following short period of oral contraceptive use. PMID- 10477199 TI - Aneurysmal bone cyst: rapid deterioration in a long-standing tumor. PMID- 10477200 TI - Birth and death: through a child's eyes. PMID- 10477201 TI - Case management in family practice: assuring cost-effective care for high-risk patients. PMID- 10477202 TI - Pediatric admissions by family physicians. PMID- 10477203 TI - Family physicians as generalists. PMID- 10477204 TI - Blood glucose control in pregnancy. PMID- 10477205 TI - Sudden death in young patients with Type 1 diabetes: a consequence of disease, treatment or both? PMID- 10477206 TI - Is undetected autonomic dysfunction responsible for sudden death in Type 1 diabetes mellitus? The 'dead in bed' syndrome revisited. AB - AIMS: Sudden nocturnal death in young persons with Type 1 diabetes mellitus has been recently described, and is known as the 'dead in bed' syndrome. Its aetiology is unknown, and we have therefore explored the details of all papers recording the syndrome, to formulate a hypothesis of causation. METHODS: Literature review of 'dead in bed' reports as well as of nocturnal hypoglycaemia, and autonomic dysfunction in relation to baroreceptor-cardiac reflex sensitivity. RESULTS: Clinical reports of 'dead in bed' cases strongly suggest that nocturnal hypoglycaemia is a likely precipitant, but that the death is sudden and probably arrhythmic. Ventricular dysrhythmias may occur in the context of early autonomic neuropathy, with relative sympathetic overactivity, in young Type 1 diabetic persons. CONCLUSION: We conclude that the 'dead in bed' syndrome probably occurs in Type 1 diabetic persons with early autonomic neuropathy, resulting in relative sympathetic overactivity. In such persons, risks of ventricular dysrhythmias will be compounded by nocturnal hypoglycaemia, which may be associated with an increase in the electrocardiographic Q-T interval, and Q-T dispersion. This could lead to the observed sudden death in undisturbed beds. Further research in this area is urgently needed, in particular into the possible protective use of drugs that modulate the autonomic nervous system. PMID- 10477207 TI - Painful diabetic neuropathy. AB - Chronic painful diabetic neuropathy causes symptoms that can last for many years and severely impair the quality of life of affected patients. This review describes the epidemiology, pathophysiology and treatment of chronic neuropathic pain. Particular emphasis is placed on a comprehensive review of the management of painful symptoms through a detailed review of the published literature using a variety of databases particularly Medline and EMBASE. PMID- 10477208 TI - Twinning project: Israel and Georgia--the birth of a diabetes-in-pregnancy centre in Georgia. AB - AIMS: To determine the effectiveness of a joint Israeli-Georgian twinning project aimed at introducing modern methods of perinatal care to Georgian women with pregestational diabetes mellitus. METHODS: A Diabetes-in-Pregnancy Centre was established in Georgia (in the former USSR). Thirty-two women with Type 1 diabetes mellitus participated in the first stage of the study (January 1997-June 1998). All were maintained under strict metabolic surveillance starting at least three months prior to conception and were given organized instruction in methods of self-monitoring of glucose levels, insulin dose adjustment, dietary management and close fetal surveillance throughout pregnancy. A second stage of the project was started in January 1998 and aimed at the diagnosis of gestational diabetes mellitus (GDM). RESULTS: Of the 32 women, 20 had had a total of 44 prior pregnancies without proper perinatal care (1990-96); only five (11.3%) ended in the birth of a healthy infant. On entry to the study, all 32 patients had unsatisfactory metabolic indices. HbA1c levels decreased significantly from the preconception period (P<0.001) and were maintained at the lower level throughout pregnancy; insulin doses decreased significantly until the third trimester (P<0.01) and then increased (P<0.001). Since January 1997, 21 women have become pregnant. Eighteen have given birth, 12 (67%) by Caesarean section and six (33%) by vaginal delivery. Gestational age at birth was 36-39 weeks, and birth weight ranged from 2300 to 4100 g. The only neonatal complications were mild respiratory distress syndrome and hypoglycaemia. There were no significant maternal complications in the 236 women screened, eight were diagnosed as having GDM and 12 impaired glucose tolerance. They were actively managed to a successful outcome. CONCLUSIONS: The establishment of the Diabetes-in-Pregnancy Centre in the Republic of Georgia has significantly reduced the prior high pre-programme perinatal morbidity and mortality as well as the incidence of maternal complications in pre-GDM as well as in GDM. PMID- 10477209 TI - Metformin treatment leads to an increase in basal, but not insulin-stimulated, glucose disposal in obese patients with impaired glucose tolerance. AB - AIMS: This study was initiated to test the hypothesis that metformin treatment leads to enhanced glucose disposal at ambient insulin concentrations. METHODS: Nineteen obese patients with impaired glucose tolerance (IGT) were treated with either metformin or placebo in a randomized, double-blind, placebo-controlled, cross-over study. Insulin secretion and insulin resistance were quantified using the homeostasis model assessment (HOMA) and insulin-stimulated glucose disposal were measured by determining the steady-state plasma glucose (SSPG). RESULTS: The average benefit of metformin was 0.6 mmol/l for glucose (95% confidence interval (CI) 0.2-0.9 P = 0.002), 2.8 pmol/l for insulin (95% CI 0.2-5.4, P = 0.019). Insulin resistance, as quantified by HOMA, was improved by 1.1 (95% CI 0.2-2.0, P = 0.004), without any change in insulin secretion. Basal and insulin-stimulated glucose oxidation were comparable in the placebo and metformin-treated groups at the end of each treatment period, as was the SSPG concentration. However, both systolic and diastolic blood pressures fell significantly following metformin administration as compared to treatment with placebo. CONCLUSIONS: These results indicate that metformin administration to patients with IGT is associated with enhanced glucose disposal at baseline insulin concentrations and a fall in blood pressure. In contrast, neither glucose oxidation nor glucose disposal were increased in association with metformin treatment under conditions of physiological hyperinsulinaemia. PMID- 10477210 TI - Relative hypoleptinaemia in women with mild gestational diabetes mellitus. AB - AIMS: There is increasing evidence suggesting that leptin plays a major role in the regulation of energy homeostasis, as well as in the neuroendocrine and reproductive systems. Leptin is synthesized in the human placenta. The aim of this study was to relate serum leptin levels during pregnancy to glucose tolerance, body mass index (BMI) and specific metabolic variables, such as specific insulin and proinsulin. METHODS: A 2-h 75 g oral glucose tolerance test was performed in 221 pregnant women at 22-29 weeks of gestation (median 25th week). Serum leptin was measured using a radioimmunoassay. In 49 women, sequential leptin measurements were performed (during pregnancy and post partum (median 5 months)). RESULTS: During pregnancy serum leptin was significantly related to body weight (r = 0.49), BMI (r = 0.51), fasting immunoreactive insulin (r = 0.46), specific insulin (r = 0.43) and proinsulin (r = 0.29) (all P-values <0.0001). In women with mild gestational diabetes (GDM, n = 55), leptin levels were lower compared to women with normal glucose tolerance (n = 166) after adjusting for BMI and fasting insulin (26.9 vs. 19.4 ng/ml, P = 0.0001). Leptin was significantly higher during pregnancy compared to post partum (mean +/- SE: 24.3+/-1.5 vs. 19.6+/-1.6 ng/ml, P = 0.0003), even after adjustment for changes in BMI and changes in fasting insulin (P = 0.013). CONCLUSIONS: Leptin levels are elevated in pregnancy. Women with mild GDM presented with relative hypoleptinaemia compared to women with normal glucose tolerance. PMID- 10477211 TI - Is glycation of low density lipoproteins in patients with Type 2 diabetes mellitus a LDL pre-oxidative condition? AB - AIMS: The study aimed to evaluate whether low density lipoprotein (LDL) in diabetic patients is more glycated and susceptible to oxidation than in non diabetic subjects and investigated the hypothesis that LDL glycation is associated with an increased plasma concentration of LDL- (a circulating electronegatively charged LDL), proposed as an index of in vivo oxidation. METHODS: LDL glycation was measured by a competitive enzyme immunoadsorbent assay, using a monoclonal antibody against glycated apoB in 24 Type 2 diabetic patients and 12 healthy controls. LDL- was separated by ion-exchange HPLC in LDL samples obtained after sequential preparative ultracentrifugation (density range 1.019-1.063). In vitro LDL susceptibility to oxidation was evaluated by following the kinetics of conjugated diene formation and by measuring the lag-phase time in the presence of copper (Cu2+) ions. RESULTS: The percentages of glycated apoB (3.33+/-2.54% vs. 1.24+/-0.71%) and of LDL- (3.88+/-1.49% vs. 2.34+/-1.03%) in total LDL were significantly higher in diabetic patients (P<0.01 for both). LDL- was positively correlated with glycated apoB (r = 0.68, P<0.001). LDL isolated from Type 2 diabetic patients showed a significant decrease (P<0.001) in the resistance to oxidative stress, as indicated by the shorter lag-phase time (91+/ 12.6 vs. 120+/-24.5 min). The lag-phase time was inversely correlated with glycated apoB (r = -0.65, P<0.001) and LDL- concentrations (r = -0.69, P<0.001). CONCLUSIONS: In this population of Type 2 diabetic patients, LDL were more glycated, more susceptible to in vitro oxidation and had a higher percentage of electronegative LDL. The glycation of apoB is proposed to be associated with a significative increase of in vivo and in vitro LDL oxidation. PMID- 10477212 TI - Gastrointestinal symptoms in Chinese patients with Type 2 diabetes mellitus. AB - AIMS: To examine and compare gastrointestinal (GI) symptoms in Hong Kong Chinese Type 2 diabetic outpatients and non-diabetic control subjects. METHODS: A total of 149 Chinese Type 2 diabetic patients (66 men and 83 women, age (mean +/- SD) 46.8+/-11.1 years) newly referred to the diabetes clinic of the Prince of Wales Hospital, Hong Kong were examined. Sixty-five age and sex-matched non-diabetic subjects were recruited from the community as controls (22 men and 43 women, age (mean +/- SD) 46.5+/-6.6 years, P = 0.820). All patients were interviewed regarding GI symptoms over the past year, using a questionnaire that covered 14 items. A scoring system from 0 to 4 was used to grade severity. RESULTS: Diabetic patients had higher blood pressure, fasting plasma glucose and glycated haemoglobin and were more often smokers than control subjects. Of the 149 diabetic subjects, 105 (70.5+/-45.8%) had GI symptoms while only 20 (30.8%) of the 65 control subjects had GI symptoms (P<0.001). The respective percentages of upper and lower GI symptoms in diabetic and normal subjects were 44.3% vs. 24.6% (P = 0.006) and 54.4% vs. 13.9% (P<0.001). The three commonest GI symptoms in diabetic patients were diarrhoea (34.9%), constipation (27.5%) and epigastric fullness (16.8%). After adjustment for age, sex, duration of diagnosed diabetes and smoking, patients with or without metformin had similar percentages or scores for GI symptoms. On multivariate analysis using age, body mass index, fasting plasma glucose, glycated haemoglobin, duration of diagnosed diabetes and presence of peripheral neuropathy as independent variables, duration of diabetes was the only independent parameter associated with total score for GI symptoms (beta = 0.116, P = 0.003), for upper GI symptoms (beta = 0.073, P = 0.005) and for lower GI symptom (beta = 0.043, P = 0.020). CONCLUSIONS: Up to 70% of the Chinese Type 2 diabetic outpatients have GI symptoms, which is a much higher rate than in non diabetic control subjects. Duration of diabetes is the most important factor associated with the presence of such GI symptoms. PMID- 10477213 TI - Oral candidal carriage and infection in insulin-treated diabetic patients. AB - AIM: To evaluate candidal load and carriage of candidal species in 414 insulin treated diabetes mellitus patients with and without clinical signs of infection. Host factors that could influence candidal load in diabetic patients with oral candidosis were also investigated. METHODS: Candidal species were recovered from 414 insulin-treated diabetes mellitus patients attending two hospital diabetic clinics, using an oral rinse technique. RESULTS: Seventy-seven per cent of diabetic patients carried Candida species in their oral cavity, with C. albicans being the species most frequently isolated. C. dubliniensis was found for the first time in this patient group. Forty per cent of patients colonized with candidal species had no clinical signs of oral candidosis. Where oral candidosis was present, erythematous candidosis was the most common clinical presentation. Candidal load was not associated with age, sex or glycaemic control. However, it was significantly increased in those patients who were tobacco smokers, and non significantly increased in those patients who wore dentures, or who had clinical signs of oral candidosis. CONCLUSION: The epidemiology of oral candidal carriage and infections in diabetic patients is complex and includes species which have not been previously reported in this group of patients. The development of oral candidosis in insulin-treated diabetes mellitus patients is not the result of a single entity, but rather, a combination of risk factors. PMID- 10477214 TI - The use of digital cameras in a mobile retinal screening environment. AB - AIMS: To assess how successfully digital camera technology might be incorporated into a mobile screening environment. METHODS: One hundred and ninety-seven people had their fundi photographed using a Topcon/Imagenet digital system and 534 using a Canon CR5/Ris-Lite system in addition to concurrent 45 degrees CR4NM Polaroid photography. One hundred and eighteen randomly selected patients were also sent for 7 field stereo photography as a gold standard. An acceptability questionnaire was answered by a random sample of those photographed. RESULTS: For the detection of any retinopathy, digital pictures had a sensitivity of 0.74 (95% confidence interval (CI) 0.68-0.80) whilst Polaroid was 0.72 (95% CI 0.66-0.78) and for referable retinopathy digital pictures had a sensitivity of 0.85 (95% CI 0.80 0.90) and Polaroid was 0.90 (95% CI 0.86-0.94). A concurrent ophthalmoscopic evaluation improved the sensitivity to 0.92 (95% CI 0.86-0.98) for detection of any retinopathy whilst the sensitivity at the referral level was improved to 0.95 (95% CI 0.91-0.99). Twenty-nine of 176 respondents experienced flash discomfort with the Polaroid system with only four of 154 describing 'some' discomfort from the digital systems which have a lower flash power (10 W vs. 300 W) and a faster recovery time. CONCLUSIONS: This study indicates that digital systems are a feasible and acceptable alternative to Polaroid-based cameras for use in a mobile environment. PMID- 10477215 TI - Do diabetes clinic attendees stay out of hospital? A matched case-control study. AB - AIMS: To examine whether routine care for diabetes mellitus influences the risk of hospital admission. METHODS: All people with diabetes in five randomly selected general practices in the city of Leicester were identified from medical records and prescribing information. Cases with a hospital admission between 1992 and 1995 but no admission in the preceding 2 years were compared with age-matched controls in a nested study. RESULTS: The variables significantly associated with an increased risk of admission were duration of diabetes in years (OR 1.07, 95% confidence interval (CI) 1.03-1.11) and number of non-diabetic drugs prescribed (OR 1.51, 95% CI 1.27-1.79). Having attended a hospital clinic in the previous 2 years was associated with reduced risk of admission (OR 0.30, 95% CI 0.14-0.65), whilst having been seen for a diabetes review in general practice was not (OR 0.91, 95% CI 0.41-1.99). Similar results were found for both diabetes-related and unrelated admissions. CONCLUSIONS: Although general practice-based review was not associated with a change in the risk of admission, attendance at a hospital clinic was associated with a decreased risk of admission. These results may be explained by the characteristics of those who attend hospital clinics, as well as by the possible effectiveness of access to specialist services in reducing admissions. PMID- 10477216 TI - Contra-indications to metformin therapy are largely disregarded. AB - AIMS: To investigate the current metformin treatment practice and in particular to examine the consideration given to its contraindications. METHODS: A cross sectional analysis of 308 consecutive Type 2 diabetic patients (mean age 66+/ 11.3 years) previously treated with metformin on an outpatient basis and admitted to a German general hospital during the period from 1 January 1995 to 31 May 1998 because of acute disease or in order to optimize their diabetes management. All patients underwent a basic investigation comprising a documentation of their medical history, a physical examination, an electrocardiogram, and an extensive laboratory profile; 34% also had acute coronary angiography. RESULTS: On admission to hospital, 73% of the patients were found to have contra-indications, risk factors, or intercurrent illnesses necessitating discontinuation of metformin; 51% of these patients had several of these conditions. As major contra indications to metformin, renal impairment was present in 19% of all patients, heart failure in 25%, respiratory insufficiency in 6.5%, and hepatic impairment in 1.3%. The risk factors to metformin included advanced coronary heart disease in 51%, atrial fibrillation in 9.8%, chronic alcohol abuse in 3.3%, advanced peripheral vascular disease in 2%, and pregnancy in 0.7%. As intercurrent illnesses, cerebral ischaemia occurred in 9.8% under metformin treatment and malignancies were diagnosed in 6.5%. The patients with contra-indications or requiring caution to metformin were significantly older and had previously been treated with more cardiovascular medication than those without such reservations (P<0.001). CONCLUSIONS: Despite the considerable risk of lactic acidosis in the majority of patients, no cases were observed. PMID- 10477217 TI - Laboratory-based calculation of coronary heart disease risk in a hospital diabetic clinic. AB - AIMS: To develop an estimation of risk of coronary heart disease (CHD) based on the Framingham equation for use in a diabetes clinic, given concerns about the accuracy of the Sheffield risk tables in this setting. METHODS: A computer program using the Framingham equation based on patients' age, sex, systolic blood pressure, smoking history, presence of diabetes and left ventricular hypertrophy was applied to requests for lipid screening of patients attending the diabetes clinics of Birmingham Heartlands Hospital. The calculated risks for the population were compared with those estimated from the Sheffield tables. RESULTS: Of 1060 patients with diabetes mellitus, 215 (20%) had an annual CHD risk > or =3%, which is considered to be the threshold at which lipid-lowering drugs are cost-effective. Only 24 of these 215 patients (11%) were correctly identified by the Sheffield tables, which we conclude have an unacceptably low sensitivity in diabetes mellitus. CONCLUSIONS: A laboratory-based CHD risk calculation system is a practical alternative to the Sheffield system and may have a greater sensitivity in the diabetic clinic. PMID- 10477218 TI - Episodic hyperglycaemia in pregnant women with well-controlled Type 1 diabetes mellitus: a major potential factor underlying macrosomia. AB - AIMS: To test the common assumption that pregnant women who are sufficiently motivated to achieve very good HbA1c levels will record home blood glucose data accurately. METHODS: A new device was used to download information from electronic blood glucose meters to assess the extent of selectivity in patient glucose diary-keeping. RESULTS: In an index case, a woman with excellent ambient HbA1c (5.9%; upper limit of normal 6.1%) was observed to have 68% of preprandial blood glucose readings above the target range of 3.5-6.5 mmol/l and a mean (+/- SD) level of 8.9+/-3.9 mmol/l in the corresponding period. No such impression was conveyed by the home monitoring diary. Six pregnant women with well controlled Type 1 diabetes (mean HbA1c 6.6+/-0.2%) exhibited between 42 and 68% of preprandial readings above the target range. CONCLUSIONS: The frequency of hyperglycaemia has hitherto been underestimated in well controlled pregnant women whose near-perfect home monitoring record is apparently corroborated by near normal HbA1c levels. These observations provide a hypothesis for understanding of the disappointing continuance of macrosomia despite excellent HbA1c levels throughout pregnancy. PMID- 10477219 TI - Adult strabismus surgery: the rehabilitation of a disability. PMID- 10477220 TI - Color doppler imaging of the central retinal artery in premature infants undergoing examination for retinopathy of prematurity. AB - PURPOSE: Recent attempts have been made to quantify blood flow velocity in the central retinal artery (CRA) of adults using color Doppler imaging (CDI). Although retinal vascular abnormalities are the hallmark of severe retinopathy of prematurity (ROP), normal values have not been established for CRA blood flow velocity in premature infants. METHODS: CDI of the CRA was successfully performed on 43 eyes in 22 infants (postconceptional ages 32 to 39 weeks) before the infants underwent examination for ROP. Peak systolic velocity (PSV) and end diastolic velocity were recorded from at least 1 eye of each patient. Pourcelot's resistive index was then calculated for each eye studied. RESULTS: Mean PSV for patients with no ROP (n = 6) was 7.2 +/- 1.5 cm/s, whereas those with any degree of ROP excluding plus disease (n = 9) had a mean PSV of 8.9 +/- 1.8 cm/s. Of the patients with ROP and plus disease (n = 7), the mean PSV was 7.0 +/- 1.6 cm/s. There were no statistically significant differences among these 3 groups (P= .08). CONCLUSIONS: CDI can be successfully performed on preterm infants and yields values lower than those previously reported in healthy adult subjects. PSV in the CRA may be higher in subjects with ROP in the absence of plus disease; however, further study is needed to determine whether these differences are significant. PMID- 10477221 TI - The mobius sequence: a relook. AB - BACKGROUND: The constellations of findings often referred to as Mobius syndrome might be better described as Mobius sequence, because the term sequence defines a cascade of secondary events after an embryonic insult from heterogeneous causes. Classic clinical findings include evidence of sixth and seventh cranial nerve involvement, often with associated malformations of limbs, craniofacial structures, and other cranial nerves. METHODS: A prospective study was undertaken in Sweden of 25 patients who showed characteristic findings of Mobius sequence. RESULTS: Of the patients who did not have strabismus surgery, 10 patients had straight eyes in the primary position, 7 had esotropia, 2 had exotropia, and 1 had hypertropia. All had significant limitation of abduction, except 1 patient with exotropia who showed minimal underaction on abduction but a large limitation of adduction. In the description in these early cases, some patients manifested a clinical pattern resembling a horizontal gaze paresis. Narrowing of the palpebral fissure on adduction similar to that seen in Duane syndrome was observed in a few cases. Two patients had ptosis. Nineteen patients had diminished facial expression bilaterally, often asymmetric, and 6 cases appeared to be unilateral. Seven patients had abnormal tearing. Associated systemic findings included Poland anomaly (2), club feet or other limb anomalies (8), micrognathia (8), tongue malformations (17), cleft palate (5), and speech problems (18). An unusual finding was autism syndrome (6) or autism-like syndrome (1). CONCLUSIONS: The associated findings in Mobius sequence may give further clues to the location and timing of the developmental disturbance. The wide range of ocular motility patterns suggests that the previous concept of a lesion solely in the sixth nerve nucleus is an inadequate explanation for these findings. PMID- 10477222 TI - The negative psychosocial impact of strabismus in adults. AB - INTRODUCTION: The goal in the treatment of strabismus is to realign the visual axes to eliminate diplopia, produce or restore binocular vision, expand the binocular visual field, and/or provide a normal appearance. The treatment of strabismus in adults who do not experience diplopia or who do not have binocular potential has sometimes been regarded as "cosmetic." Many adults with strabismus have stated that it has had a negative effect on their lives. This study was designed to ascertain whether there is a negative social bias against adult patients with strabismus that could affect interpersonal relationships and limit employment opportunities. METHODS: An orthotropic person was photographed in primary, right, and left gaze. These photographs were then digitally altered to produce an esotropia and exotropia of similar magnitude. Subjects were then asked to evaluate a single photograph with reference to personality characteristics that are important for social interaction and employment capability. RESULTS: Overall, the strabismic faces were judged significantly more negatively, across 11 descriptive characteristics, than the nonstrabismic face. The effect of esotropia was worse than exotropia. CONCLUSION: Strabismus creates a significant negative social prejudice. These biases can have a detrimental impact on socialization and employability. Treatment of strabismus may positively alter perceived characteristics of individuals and improve their ability to socialize normally and obtain employment. Therefore, the treatment of strabismus should not be considered cosmetic even when there is no hope of improving binocular vision. PMID- 10477223 TI - The clinical and morphologic spectrum of optic nerve hypoplasia. AB - PURPOSE: The purpose of this study was to characterize the clinical and morphologic spectrum of all children referred for optic nerve hypoplasia to a tertiary referral hospital in Sweden during a 9-year period. SUBJECTS AND METHODS: A retrospective review was undertaken of the charts of 117 children (age range, 0.25-16 years), treated at the Children's Hospital, Goteberg between 1988 and 1996, after the diagnosis of optic nerve hypoplasia. Ocular fundus morphologic condition was evaluated by digital image analysis of fundus photographs in 50 children, and neuroimaging was performed in 57 children. RESULTS: Of the 117 children with optic nerve hypoplasia, 66 (56%) were boys and 51 (44%) were girls. Preterm birth occurred in 24 (20%), and 14 (12%) were born small for gestational age. Additional diagnoses, such as fetal alcohol syndrome, septo-optic dysplasia, perinatal adverse events, and neuropsychiatric disorders, were made in 88%; 7% had unilateral optic nerve hypoplasia. Most of the children had small optic disc, cup, and neuroretinal rim areas, as well as retinal vascular abnormalities; 75% were visually impaired, and a high incidence of nystagmus and strabismus was found among these children. CONCLUSION: This study indicates that optic nerve hypoplasia has a wide clinical and morphologic spectrum and is associated with a broad range of disorders of the central nervous system. It is suggested that differences in the etiology and timing of the lesion as well as associated lesions may explain this spectrum of optic nerve hypoplasia in children. PMID- 10477224 TI - The relationship between stereopsis and monocular optokinetic optokinetic nystagmus after infantile cataracts. AB - PURPOSE: Visual deprivation disrupting binocular development, such as that occurring with congenital cataract, is reported to cause asymmetric monocular optokinetic nystagmus (MOKN), as well as poor sensory and motorfusional outcome. We wanted to determine if symmetric MOKN could develop in cases of congenital cataract with good fusional outcome. METHODS: We tested MOKN (with video and electro-oculographic recordings) and stereoacuity on 5 patients with good visual acuity and satisfactory ocular alignment after surgery for congenital cataract. RESULTS: Stereoacuity was better than 50 seconds of arc in 1 case of monocular cataract and 2 cases of bilateral cataract. These case patients had symmetric MOKN. In a monocular cataract case, symmetric MOKN was observed in spite of questionable stereoacuity (at least 500 arc/s). One patient showed asymmetric MOKN, despite good visual acuity, and stereoacuity of 200 arc/s. CONCLUSIONS: Patients with congenital cataract can have symmetric MOKN and good stereopsis. These cases suggest that MOKN symmetry develops along with good stereopsis, but the quality of stereopsis necessary for development of MOKN symmetry remains unclear. PMID- 10477225 TI - Stereopsis after primary in-the-bag posterior chamber implantation in children. AB - PURPOSE: The purpose of this study was to evaluate visual acuity and binocular function after primary posterior chamber intraocular lens (IOL) implantation in children. PATIENTS AND METHODS: A retrospective chart review of 39 eyes of 31 children was performed. Fifteen eyes with traumatic, 17 with developmental, and 7 with congenital cataracts without any other ophthalmologic problems were examined before and after cataract surgery (irrigation/aspiration procedure with implantation of a posterior chamber IOL). Twelve eyes also received a posterior capsulorrhexis and anterior vitrectomy. The mean age at surgery was 6.9 +/- 3 years (range, 3-12 years). RESULTS: Twenty (51%) of 39 eyes achieved a best corrected postoperative visual acuity of 20/40 or better (range, 20/200-20/20). The mean postoperative visual acuity was 20/40 in the traumatic and developmental cataract groups and 20/100 in the congenital cataract group. There was a positive correlation between cataract morphology and visual acuity (P<.05). Bilateral cataracts had a better postoperative visual acuity than unilateral cataracts (P <.005). Nineteen (70%) of 27 eyes in which no primary posterior capsulorrhexis had been performed had posterior capsule opacification. Stereopsis was found in 10 of the 31 patients: 43% of the traumatic cataract group, 30% of the developmental cataract group, and 14% of the congenital cataract group. CONCLUSIONS: After capsular bag-fixated IOL, visual acuity and binocular function in children older than 3 years were favorable and the complication rate, excluding posterior capsule opacification, was low. PMID- 10477226 TI - Therapeutic outcomes of cryotherapy versus transpupillary diode laser photocoagulation for threshold retinopathy of prematurity. AB - INTRODUCTION: We undertook this study to compare the clinical outcomes in patients treated for threshold retinopathy of prematurity (ROP) with transpupillary diode laser photocoagulation versus transscleral cryotherapy. METHODS: A retrospective chart review was performed of patients treated for threshold ROP at our institution between 1988 and 1997. Cryotherapy was used to treat patients before 1992, and diode laser was used thereafter. One hundred fifteen eyes of 63 patients underwent cryotherapy, and 130 eyes of 70 patients underwent laser treatment. Because strong concordance exists between fellow eyes treated for ROP, statistical analysis was done for right eye outcomes only. Demographics, short-term complications, and treatment parameters were compared on the entire cohort of patients. Structural outcomes were compared between a group of 79 cryotreated eyes (39 OD, 40 OS) and a group of 113 laser-treated eyes (56 OD, 57 OS). Mean cycloplegic refraction was compared between a group of 38 cryotreated eyes (18 OD, 20 OS) and a group of 90 laser-treated eyes (46 OD, 44 OS). Visual acuity for preverbal and nonverbal children was estimated and converted to a Snellen visual acuity equivalent. Geometric mean visual acuity was then compared between a group of 44 cryotreated eyes (22 OD, 22 OS) and a group of 92 laser-treated eyes (47 OD, 45 OS). Patients were included in each outcome analysis if adequate documentation was present in the patient record. RESULTS: Demographics and short-term complications were not significantly different between the 2 groups. In the statistical group, 22 of 39 right eyes (56.4%) in the cryotherapy group versus 49 of 56 right eyes (87.5%) in the laser group had resolution of ROP after treatment (P = .0008). Mean spherical equivalent cycloplegic refraction at 12 months of age and over the follow-up period was not significantly different between the cryotherapy and laser groups. Estimated geometric mean visual acuity in the cryotherapy group was 20/103 and in the laser group was 20/49 at 12 months of age (P= .0099). CONCLUSIONS: Diode laser photocoagulation was associated with a better long-term structural outcome and visual acuity compared with cryotherapy for the treatment of threshold ROP. Refractive error was not significantly different between the 2 groups over an equivalent follow-up period. PMID- 10477227 TI - Visual acuity after cycloplegia in children: implications for atropine penalization. AB - BACKGROUND: Atropinization of the sound eye is an alternative to patching in the treatment of amblyopia. Whether atropine treatment can induce a switch in fixation depends on the refractive error of the sound eye, visual acuity of the amblyopic eye, distance from the fixation target, and presence of any optical correction or penalization. General guidelines are needed on the basis of refractive error and visual acuity in the amblyopic eye to predict which patients may potentially benefit from atropine penalization. METHODS: Refractive error and visual acuity at distance (6 m) and/or at near (33 cm) were recorded in a normal eye of 126 consecutive children (mean age, 8.2 years), 30 to 60 minutes after receiving cyclopentolate 1%. Visual acuity was plotted versus refractive error at distance and at near, and best-fit curves were calculated. RESULTS: There was a consistent, reproducible relationship between refractive error and visual acuity after cycloplegia at both distance and near in healthy children. CONCLUSIONS: The results of this study can be used to quickly determine whether atropine penalization has the potential for success on the basis of a patient's visual acuity in the amblyopic eye and refractive error in the sound eye. When adequate hyperopia is present in the sound eye, one should consider testing for fixation preference or initiating a therapeutic trial of atropine. Those children with insufficient hyperopia in the sound eye relative to visual acuity in the amblyopic eye can be spared the time, expense, and potential side effects of atropine penalization. PMID- 10477228 TI - Incidence of presumed cytomegalovirus retinitis in HIV-infected pediatric patients. AB - INTRODUCTION: Large population studies of adult patients suggest an incidence of cytomegalovirus (CMV) retinitis as high as 19% to 20% as a late complication of adult HIV infection. We conducted this prospective study of a large cohort of HIV infected children to determine the incidence of CMV retinitis in HIV-infected children. METHODS: From January 1984 to August 1997, 173 HIV-infected children were followed up for an average of 55.3 months (13-164 months). The patients were seen in the Department of Pediatrics at least once every 6 months. Ophthalmologic examinations were initiated when a patient's CD4 count dropped below 50 or sooner if required for ophthalmologic or other indications. Ophthalmologic examination was then repeated every 6 months. RESULTS: A total of 116 (67%) of 173 patients underwent ophthalmologic examination. Four (3.4%) of 116 patients had CMV retinitis at a mean time of 17.3 months (8-38 months) after their CD4 counts dropped below 20. None of the 4 patients with CMV retinitis had subjective visual complaints despite advanced retinitis. Three patients had bilateral and 1 patient had unilateral CMV retinitis. CONCLUSIONS: CMV retinitis occurred infrequently in HIV-infected pediatric patients and was diagnosed only in patients with a CD4 count below 20. Routine ophthalmologic screening examinations may not be necessary in pediatric patients until the CD4 count is below 20. Because children may not complain of decreased vision, at-risk children should undergo frequent ophthalmologic examination. PMID- 10477229 TI - Successful treatment of tractional corectopia using 2 mJ of energy with an Nd:YAG laser. AB - Although the Nd:YAG laser is most commonly used to perform posterior capsulotomies after cataract surgery, it has also been used to treat a variety of other anterior segment abnormalities including tractional corectopia, iridocorneal adhesions, persistent pupillary membranes, and posterior synechiae. Numerous reports on the use of the Nd:YAG laser to treat structures in the anterior segment have emphasized the need to use higher pulse energy for pupillary membranes, compared with the lower settings required for posterior capsulotomy. Steinert and Puliafito noted that single pulses of 4 to 12 mJ may be required to treat pupillary membranes "in a manner similar to that of a stonemason chipping at marble" in their description treating a membrane considerably thicker than what we describe. We report the successful treatment of tractional corectopia due to an anterior membrane strand in a child with only 2 mJ of total energy. PMID- 10477230 TI - Subretinal lesions in subacute sclerosing panencephalitis. AB - Subacute sclerosing panencephalitis (SSPE) is a rare progressive neurologic disorder. A 9-year-old boy was seen who had progressive neurocognitive decline, myoclonic jerking of the extremities, and an abnormal result of an electroencephalogram (EEG). Ophthalmoscopic examination revealed multifocal subretinal lesions. The diagnosis of SSPE was made on the basis of the clinical examination and elevated serum and spinal fluid measles titer. We describe subretinal lesions in a patient with SSPE. PMID- 10477231 TI - Bilateral vitreous hemorrhages in an infant with low fibrinogen levels. AB - The finding of retinal or vitreous hemorrhage in a child under age 3 years may cause significant controversy with regard to the etiology, because it raises the suspicion of nonaccidental injury. Blood dyscrasias have been documented to cause retinal and vitreous hemorrhages in adults and children, but they have rarely been reported to be the cause of retinal hemorrhages in neonates. We report on a patient with a low plasma fibrinogen level who had bilateral retinal hemorrhage that proceeded to vitreous hemorrhage. This subtle abnormality of the blood clotting cascade caused significant retinal and vitreous hemorrhage in a child without risk factors for abuse. PMID- 10477232 TI - Vertical latent nystagmus component and vertical saccadic asymmetries in subjects with dissociated vertical deviation. PMID- 10477233 TI - The neurology of traumatic "dissociative" amnesia: commentary and literature review. AB - BACKGROUND: The relationship between traumatic emotional stress, hippocampal injury, memory loss, and traumatic ("dissociative") amnesia was examined. METHOD: A survey of the research on emotional trauma, learning, memory loss, glucocosteroid stress hormones, and the hippocampus was conducted, and animal and human studies were reviewed. RESULTS: It is well documented and has been experimentally demonstrated in animals and humans that prolonged and high levels of stress, fear, and arousal commonly induce learning deficits and memory loss ranging from the minimal to the profound. As stress and arousal levels dramatically increase, learning and memory deteriorate in accordance with the classic inverse U-shaped curve. These memory deficits are due to disturbances in hippocampal activation and arousal, and the corticosteroid secretion which can suppress neural activity associated with learning and memory and induce hippocampal atrophy. Risk and predisposing factors include a history of previous emotional trauma or neurological injury involving the temporal lobe and hippocampus, the repetitive and prolonged nature of the trauma, and age and individual differences in baseline arousal and level of cortisol. CONCLUSIONS: Although some victims may be unable to forget, amnesia or partial memory loss is not uncommon following severe stress and emotional trauma. Even well publicized national traumas may induce significant forgetting. Memory loss is a consequence of glucocosteroids and stress-induced disturbances involving the hippocampus, a structure which normally plays an important role in the storage of various events in long-term memory. PMID- 10477234 TI - Predicting child maltreatment recurrences during treatment. AB - The primary purpose of the child protective services system is to protect children from the recurrence of child maltreatment. Understanding more about what predicts recurrence may help us more adequately target interventions to reduce the risk of future maltreatment. OBJECTIVE: The specific objective of this study was to identify correlates of recurrence during CPS intervention for families who were provided continuing intervention following a confirmed index report of physical abuse or neglect. METHOD: This nonconcurrent prospective study selected 446 subject families who met study eligibility requirements from 1,181 families randomly selected from the 2,902 families who had experienced a substantiated report of child abuse or neglect during the sampling year. Data were collected and coded from archival sources for 5 years following the index report. Each record was coded by two research analysts to increase inter-rater reliability. Data were analyzed with survival analysis methods: (1) Kaplan Meier and (2) the Cox Proportional Regression Model. RESULTS: Predictors of recurrence were child vulnerability, family stress, partner abuse, social support deficits, and an interaction between family stress and social support deficits. CONCLUSIONS: Implications of this and earlier research suggest that increasing social supports may help families cope with life events that increase stress and the risk of continued child maltreatment; that collaborations between CPS and domestic violence agencies are needed; and that screening maltreated children for mental health problems and other disabilities and assuring that children with these needs and their families get effective treatment may reduce the likelihood of continued maltreatment. PMID- 10477235 TI - Common errors of reasoning in child protection work. AB - OBJECTIVE: Repeated public inquiries into child abuse tragedies in Britain demonstrate the level of public concern about the services designed to protect children. These inquiries identify faults in professionals' practice but the similarities in their findings indicate that they are having insufficient impact on improving practice. This study is based on the hypothesis that the recurrent errors may be explicable as examples of the typical errors of human reasoning identified by psychological research. METHODS: The sample comprised all child abuse inquiry reports published in Britain between 1973 and 1994 (45 in total). Using a content analysis and a framework derived from psychological research on reasoning, a study was made of the reasoning of the professionals involved and the findings of the inquiries. RESULTS: It was found that professionals based assessments of risk on a narrow range of evidence. It was biased towards the information readily available to them, overlooking significant data known to other professionals. The range was also biased towards the more memorable data, that is, towards evidence that was vivid, concrete, arousing emotion and either the first or last information received. The evidence was also often faulty, due, in the main, to biased or dishonest reporting or errors in communication. A critical attitude to evidence was found to correlate with whether or not the new information supported the existing view of the family. A major problem was that professionals were slow to revise their judgements despite a mounting body of evidence against them. CONCLUSIONS: Errors in professional reasoning in child protection work are not random but predictable on the basis of research on how people intuitively simplify reasoning processes in making complex judgements. These errors can be reduced if people are aware of them and strive consciously to avoid them. Aids to reasoning need to be developed that recognize the central role of intuitive reasoning but offer methods for checking intuitive judgements more rigorously and systematically. PMID- 10477236 TI - Street children: a comparative perspective. AB - OBJECTIVE: This paper examines the findings from recent studies of street children in Ethiopia. METHODS: Following a discussion of the term "street children," comparisons are drawn between Latin American and Ethiopian street children in terms of gender, age, reasons for going to the streets, family relations and structure, delinquency, drug use, groups and the outcomes of street life. In particular, the victimization of street children in Ethiopia is examined. RESULTS: Widespread abuse of street children was reported. More than half of the street boys questioned reported being "regularly" physically attacked. Street life is also highly victimogenic for street girls. Sexual offences, in particular, were widespread. Forty four percent had been raped and a further 26% had been sexually attacked. CONCLUSIONS: Similarities between Latin American street children and their Ethiopian counterparts regarding gender, background and street life experiences are noted. Comparisons concerning the victimization of street children were not possible, as this is an issue that is relatively unexamined in the Latin American context. Also, issues such as the developmental outcomes of street life and the process of leaving street life remain to be examined. PMID- 10477237 TI - Are researchers bound by child abuse reporting laws? AB - OBJECTIVE: To discuss issues concerning mandatory reporting of child abuse in research settings. METHOD: An overview of existing Federal and State statutes regarding mandatory reporting of child abuse is presented. A critical review of the literature addresses the following issues: (1) whether researchers have a moral duty to place the health and safety of children above concerns about confidentiality and the benefits of obtaining new knowledge; (2) whether the Certificate of Confidentiality preempts reporting requirements; (3) whether researchers who are not health professionals (such as child developmentalists, psychobiologists, neuroscientists) should be required to report; and (4) whether researchers should be required to expand their protocols to include more in-depth investigation of potential abuse. RESULTS: Existing child abuse reporting laws do not specifically designate researchers as among the category of individuals mandated to report suspected child abuse. Currently, Human Subject Protection Committees and Federal funding agencies are tending to interpret reporting laws as applying to researchers, including requiring that research subjects are informed of this responsibility in consenting procedures. It is unclear whether the Certificate of Confidentiality preempts child abuse reporting laws. CONCLUSION: The authors recommend that legislatures specifically designate researchers as mandated reporters to ensure more uniform reporting practices in research settings. For both investigators and Human Subject Protection Committees, inclusion of researchers among the categories of those mandated to report would also help address issues of immunity from civil and criminal liability for "good faith" reports that turn out to be false and injurious. PMID- 10477238 TI - Typology and correlates of sexual abuse in children and youth: multivariate analyses in a prevalence study in Geneva. AB - OBJECTIVE: Child sexual abuse (CSA) encompasses a variety of distinct situations, which should be differentiated. There is a need for a working classification, taking into account various parameters. METHODS: The Geneva CSA prevalence study was based on 1116 randomly selected adolescents aged 14 to 16. Secondary analyses of the 252 reported cases of abuse was carried out using a multivariate approach combining multiple correspondence analysis and automatic classification, leading to descriptive analyses and to a new classification of CSA. RESULTS: Global prevalence of CSA, using a broad definition, was 10.9% for boys, and 33.8% for girls. Classification of cases of abuse differentiated 5 classes: (A) exhibitionism or other abuse without contact, by an adult (proportion of cases in boys: 22%; girls: 32%); (B) abuse of an adolescent boy by a peer without contact (62%); (C) abuse of an adolescent girl by a peer with physical contact (38%); (D) abuse inside the family, repeated (boys: 10%, girls: 16%); (E) child abuse with genital penetration, lasting 2 years or more (boys: 7%; girls: 14%). CONCLUSIONS: CSA is a complex and heterogeneous problem. Reported abuse situations can be classified into five classes, with different degrees of severity, causes, and consequences. Classification is essential in clinical research as well as in the design and conception of prevention programs. PMID- 10477239 TI - Childhood abuse recollections in a nonclinical population: forgetting and secrecy. AB - OBJECTIVE: This study investigated the relationship of interrupted memories of childhood abuse with the secrecy of the abuse. METHODOLOGY: Fifteen hundred people were randomly selected from the membership of the American Counseling Association and sent a questionnaire regarding childhood abuse history. Four hundred and twenty-three usable questionnaires were returned and analyzed. RESULTS: Thirty-two percent of the sample reported childhood abuse. Fifty-two percent of those reporting abuse also noted periods of forgetting some or all of the abuse. On the two survey items assessing secrecy, 76% of respondents reporting childhood abuse indicated there had been a time when no one but themselves and their abuser knew about the abuse; 47% indicated that an abuser tried to get them to keep the abuse secret. Forty percent endorsed both secrecy items. Respondents who reported forgetting abuse also reported one or both elements of secrecy more frequently than those who reported continuous memories of abuse. CONCLUSION: These findings are consistent with those of other studies that suggest that, among adults reporting childhood abuse, the experience of forgetting some or all abuse is common. Secrecy of the abuse appears to be associated with the experience of forgetting childhood abuse for many individuals. PMID- 10477240 TI - Psychologists' beliefs about the prevalence of childhood sexual abuse: the influence of sexual abuse history, gender, and theoretical orientation. AB - OBJECTIVE: This study examined the influence of sexual abuse history, gender, theoretical orientation, and age on beliefs about the prevalence of childhood sexual abuse among clinical and counseling psychologists. METHOD: A mail survey design was used in this study. Participants were randomly selected from the American Psychological Association membership database. There were 615 psychologists who completed self-report measures on beliefs about the prevalence of childhood sexual abuse and demographic characteristics. RESULTS: Overall, clinicians' scores on the prevalence of childhood sexual abuse were moderate. There were significant gender differences on beliefs, suggesting that women were more likely believe that childhood sexual abuse is a common occurrence compared to men. Multiple regression analysis indicated that clinician characteristics (history of sexual abuse, gender, and theoretical orientation) were significantly related to beliefs about the prevalence of childhood sexual abuse. However, these characteristics only accounted for a small amount of the overall variance predicting beliefs. CONCLUSIONS: These results suggest that clinicians do not hold extreme beliefs regarding the prevalence of childhood sexual abuse. Moreover, certain clinician characteristics are associated with their beliefs, which in turn, may impact their clinical judgment and treatment decisions. Furthermore, much of the variance was unaccounted for in the model indicating that psychologists' beliefs are complex and are not unduly influenced by their personal characteristics. Implications for clinical practice and future research are discussed. PMID- 10477241 TI - The timing of academic difficulties among maltreated and nonmaltreated children. AB - OBJECTIVE: In this study we examined the relationship between child maltreatment and the timing of academic difficulties. METHOD: This study uses survival analysis to investigate the timing of risk of experiencing an academic difficulty for the first time. Three types of academic difficulties were examined-grade repetitions, poor English grades, and poor math grades. The sample included approximately 300 maltreated and 300 nonmaltreated children aged 5-18. RESULTS: Maltreated children displayed greater risk than nonmaltreated children of repeating a grade and receiving a poor English and mathematics grade for the first time across most elementary years. Maltreated and nonmaltreated children share the same peaks in risk: first grade for their first grade repetitions and kindergarten for their first poor English and math grade. The best-fitting discrete time hazards models suggested underlying temporal patterns of risk vary according to the type of academic difficulty. For instance, maltreated children were at substantially higher risk than nonmaltreated children of repeating kindergarten and first grade. From second through sixth grade, maltreated and nonmaltreated children were indistinguishable in their risk of repeating a grade for the first time. In contrast, discrete-time hazards modeling showed that while the absolute risk of receiving a poor English or mathematics grade changes across the elementary years, the relative risk by maltreatment status does not. CONCLUSIONS: While maltreated and nonmaltreated children share the same peaks in risk, the relative risk changes across time for grade repetitions but not for the first occurrence of a poor English or mathematics grade. In summary, this study highlights the importance of time in understanding the relationship between child maltreatment and academic difficulties. PMID- 10477242 TI - Concerns about allegations of child sexual abuse against teachers and the teaching environment. AB - OBJECTIVE: The major objective was to determine teachers' awareness of the potential for child abuse allegations against themselves and the effects on the teaching environment. METHOD: Of 3,000 questionnaires sent out to a random sample of New York state teachers, 515 were returned. The responding teachers were similar to New York state teachers in terms of gender, age, and racial/ethnic distribution. RESULTS: A large percent of the respondents (56%) were aware of false allegations made against a teacher in their school district. About a third (36.5%) expressed concern that a child abuse allegation could be made against them. In response to a vignette, forty-two (42%) advised a new teacher against being alone in a room with a student; 62% advised against casual touching; 70% advised against hugging or putting an arm around a student. Males more than female teachers, especially those teaching upper grades, advised against such contact. The more teachers expressed concern about abuse allegations against themselves, the more teachers advised against contact. CONCLUSIONS: Fear of abuse allegations are salient for teachers. Fears may cause teachers to limit contact with students with potentially adverse consequences for students and the teaching environment. PMID- 10477243 TI - Fish consumption and coronary heart disease mortality. A systematic review of prospective cohort studies. AB - OBJECTIVES: To review all prospective cohort studies examining the relationship between fish intake and coronary heart disease mortality, and to assess the strength and consistency of their findings. DESIGN: Systematic review of studies based on individual records of fish or n-3 polyunsaturated fatty acid consumption and coronary heart disease death. Studies were given scientific quality scores and divided into categories of high, intermediate, or insufficient quality. MAIN OUTCOME MEASURE: Coronary heart disease mortality. RESULTS: Eleven studies were identified. The cohorts counted a total of 116764 individuals. Of four studies judged to be of high quality, the two largest (n = 44895 and 20051) were performed in populations at low risk of coronary heart disease. They found no protective effect of fish consumption. The other two high-quality studies were relatively small (n = 852 and 1822) and included individuals at higher risk. They both found an inverse relationship between fish consumption and coronary heart disease death, suggesting that 40-60 g fish per day is optimal and associated with a risk reduction of 40-60%. Results of four studies of intermediate quality support that fish consumption is inversely associated with coronary heart disease mortality in high-risk populations only. Three studies were judged to be of insufficient quality to be used for drawing conclusions. CONCLUSIONS: Fish consumption is not associated with reduced coronary heart disease mortality in low-risk populations. However, fish consumption at 40-60 g daily is associated with markedly reduced coronary heart disease mortality in high-risk populations. The underlying biochemical mechanism is not known and causal inference premature. PMID- 10477244 TI - Differences between polyunsaturated fatty acid status of non-institutionalised elderly women and younger controls: a bioconversion defect can be suspected. AB - OBJECTIVE: To evaluate the polyunsaturated fatty acid (PUFA) status in non institutionalised elderly women and to detect a possible essential fatty acid bioconversion defect. DESIGN AND SUBJECTS: The fatty acid composition of total plasma lipids, plasma triglycerides (TG), cholesterol esters (CE), phospholipids (PL), and erythrocytes was determined by capillary column gas-liquid chromatography in a sample of 200 non-institutionalised healthy elderly women over 75 years of age. The data were compared with those of a control group of 50 young female volunteers aged 20-48 y. RESULTS: In elderly women, the n-6 series precursor, linoleic acid (18:2 n-6), was lower in TG and CE (P = 0.029 and 0.014, respectively). In CE, this fatty acid was highly correlated with vegetable and vegetal fat intakes (P < 0.0001), suggesting a lower dietary supply than in controls. Higher percentages of 16:1 n-7 were found in all the plasma lipid fractions in elderly women, especially in CE (P < 0.0001). The ratios 20:4 n 6/20:3 n-6 and 22:6 n-3/20:5 n-3 were significantly lower in PL from elderly women (P < 0.005 and P < 0.002, respectively), raising the question of the efficiency of the terminal steps of 20:4 n-6 and 22:6 n-3 biosynthesis. Dietary investigations in elderly women indicated that a high dietary protein intake via meat probably contributed to the supply of 20:4 n-6 and thus maintained the status of this fatty acid, despite the suspected altered biosynthesis. CONCLUSION: The PUFA status in the elderly women group could be more fragile and dependent on exogenous supply of long-chain PUFAs than previously suspected. PMID- 10477245 TI - Effect of synthetic triglycerides of myristic, palmitic, and stearic acid on serum lipoprotein metabolism. AB - OBJECTIVES: To determine relative effects of diets high in synthetic sources of myristic (14:0), palmitic (16:0) or stearic (18:0) acid on concentrations and metabolism of serum lipoproteins. DESIGN: Eighteen healthy women participated in a three-way cross-over study for five week periods separated by seven week washout periods, diets were assigned in random order. SUBJECTS: Premenopausal women, not on medication, were from three races (Caucasian, African-American, Asian) and four apolipoprotein E phenotype groups (3/3, 3/2, 4/3, and 4/2). INTERVENTION: During the first week the subjects consumed a baseline diet providing 11 energy (en)% saturated fat, 10en% polyunsaturated fat and 14en% monounsaturated fat. Followed by test diets with 19en% saturated fat (including 14en% test saturated fatty acid), 3en% polyunsaturated fat, and 14en% monounsaturated fat for four weeks. Synthetic fats (trimyristin, tripalmitin, and tristearin) were used in blends with natural fats and oils. RESULTS: Mean concentrations of serum total, esterified and LDL cholesterol were significantly lower after 18:0 than after 16:0 (n = 16-18, P < 0.01 for treatment effect). Myristic acid (14:0) had an intermediate effect. Receptor-mediated degradation of 125I-LDL in mononuclear cells obtained from the subjects was lower after 16:0 than after 14:0 and 18:0 (n = 16-18, P=0.05 for treatment effect). Differences in the digestibilities of the fats were not a major factor in the results. Strong cholesterolemic responses to the 16:0 diet were partly explained by apoE phenotype. CONCLUSIONS: As noted previously, stearic acid was neutral compared to 14:0 and 16:0. In contrast to studies involving natural fats, 14:0, fed as a synthetic triglyceride, was less cholesterolemic than 16:0 in a majority of subjects. ApoE phenotype influenced the cholesterolemic response particularly when diets high in 16:0 were eaten. PMID- 10477246 TI - Age and health indications assessed by silhouette photographs. AB - OBJECTIVE: To test the hypothesis that overweight and central fat distribution make people look older and have poorer health. DESIGN: We asked 201 male and 161 female observers aged 28-67 y with body mass index (BMI) 17-45 kg/m2, to estimate the age of eight silhouette photographs of female volunteers with known BMI and waist and hips. Silhouette photographs were presented individually and grouped in three trios for analysis: trio A, each subject had waist to hip ratio = 0.7, BMI for each subject was 22, 24, 29 kg/m2; trio B, each subject had BMI = 22 kg/m2, waist to hip ratio for each subject was 0.66, 0.73, 0.99; and trio C, each subject had BMI = 29 kg/m2, waist to hip ratio for each subject was 0.76, 0.88, 0.99. The observers were then asked to judge the likelihood of developing heart disease, health and longevity of the silhouettes which were presented in four pairs controlling either for BMI or waist to hip ratio. RESULTS: The age of a silhouette with BMI 22 kg/m2 was estimated to be 35 y, 24 kg/m2 to be 45 y, and 30 kg/m2 to be 55 y, when waist to hip ratio was matched at 0.7. When BMI was controlled at either 22 or 29 kg/m2, higher waist to hip ratio led to a greater age estimation. An increase from a 65 cm to 88 cm waist circumference led to an estimate of 25 y older. Narrow hips was also judged as older. The silhouettes judged the more likely to be healthy and live longer were those with the lower waist to hip ratio (by 70% observers), and the lower BMI (by 90% observers). Waist to hip ratio appeared less influential when BMI of the silhouettes was high. The observers' own BMI, age or sex had little influences on their assessments of the age and health status of the silhouette photographs. CONCLUSIONS: Using novel sets of silhouette photographs, it was shown that overweight or central fat distribution and narrow hips suggest a person is older and has poorer health: People with BMI 29.7 kg/m2 appear to be 15-18 y older than those with BMI 22.2 kg/m2, each extra cm on the waist makes women appear to look a year older, and progressively less healthy. BMI and age of the observers had little influences on the findings. PMID- 10477247 TI - The association of diet and thrombotic risk factors in healthy male vegetarians and meat-eaters. AB - OBJECTIVE: The aim of this study was to assess thrombosis tendency in subjects who were habitual meat-eaters compared with those who were habitual vegetarians. DESIGN: Cross-sectional comparison of habitual meat-eaters and habitual vegetarians. SETTING: Free living subjects. SUBJECTS: One hundred and thirty-nine healthy male subjects (vegans n = 18, ovolacto vegetarians n = 43, moderate-meat eaters n = 60 and high-meat-eaters n = 18) aged 20-55 y who were recruited in Melbourne. OUTCOME MEASURES: Dietary intake was assessed using a semi quantitative Food Frequency Questionnaire. The parameters of thrombosis were measured by standard methods. RESULTS: Saturated fat and cholesterol intakes were significantly higher and polyunsaturated fat (PUFA) was significantly lower in the meat-eaters compared with vegetarians. In the meat-eaters, the platelet phospholipids AA levels were significantly higher than in the vegetarians, but there was no increase in ex vivo platelet aggregation and plasma 11-dehydro thromboxane B2 levels. Vegetarians, especially the vegans, had a significantly increased mean collagen and ADP stimulated ex vivo whole blood platelet aggregation compared with meat-eaters. The vegan group had a significantly higher mean platelet volume than the other three dietary groups. However, meat-eaters had a significantly higher cluster of cardiovascular risk factors compared with vegetarians, including increased body mass index, waist to hip ratio, plasma total cholesterol (TC), triacylglycerol and LDL-C levels, ratio of TC/HDL-C and LDL-C/HDL-C and plasma factor VII activity. CONCLUSIONS: Consumption of meat is not associated with an increased platelet aggregation compared with vegetarian subjects. PMID- 10477248 TI - Milk hypersensitivity in young adults. AB - OBJECTIVE: To estimate the prevalence of milk hypersensitivity in Finnish adults. DESIGN: Cross-sectional study. SUBJECTS: Two hundred men and 206 women aged 27 y randomly recruited from the population register in southwestern Finland. INTERVENTIONS: The subjects were interviewed about their dairy product consumption, abdominal discomfort after dairy product intake and lactose intolerance. From serum samples, serum reactivity to milk protein and milk specific IgG1, IgG2, IgG3 and IgA were measured. RESULTS: About 20% of the subjects reported abdominal discomfort after dairy product intake, whereas only 6.4% had been diagnosed to have lactose intolerance. The amount of milk consumed correlated well with the serum assay results in subjects reporting abdominal discomfort but not in subjects who were free from these symptoms. Among subjects with no record of dairy product restriction or lactose intolerance, those experiencing abdominal discomfort after dairy product intake had significantly higher serum reactivity to milk protein than those without such discomfort. The concentrations of serum milk-specific antibodies did not differ between these two groups. The prevalence of milk hypersensitivity in this population was estimated to be 3-6%. CONCLUSIONS: Milk hypersensitivity may be as common in adults as in infants. The measurement of serum reactivity to milk protein may prove useful in screening milk hypersensitivity in subjects who have not restricted their dairy product consumption. PMID- 10477249 TI - Within- and between-person variation of nutrient intakes of older people in Korea. AB - OBJECTIVES: (i) To examine the components of variation in nutrient intakes for older Korean adults; (ii) To calculate the level of accuracy with varying degree of precision in order to achieve estimates of usual nutrient intakes. DESIGN: Five or six 24 h recalls were made over a four month period on each individual. SETTING: A senior school from a middle-income, urban community in Inchon, Korea. SUBJECTS: Forty-seven males and 36 females over 60 y old with complete records for at least five dietary recalls. RESULTS: The within-person variation of nutrient intakes (21.8-103.2%) was higher and differed more markedly by nutrients than the between-person variation (9.4-63.7%). Males (2.26-11.28) showed larger within-to-between person variance ratios than females (1.73-5.86). The variance ratios were different by nutrients for males, but no such pattern was observed for females. Depending on nutrients, 5-6 recalls over a four month period gave estimates of nutrient intakes within 30-50% of usual intakes. For most nutrients, 100-200 subjects were sufficient to obtain estimates with 10% deviation of group usual intakes. Achieving r=0.9 between observed and usual intakes required 24-54 recalls for males and 8-23 recalls for females. CONCLUSIONS: The degree of precision achieved during assessment of older Korean adults' usual nutrient intakes changes with gender and by the type of nutrients. Achieving similar level of accuracy of the estimates, older Korean adults may require more dietary recalls than the adults in western countries. PMID- 10477250 TI - Retinol, alpha-tocopherol and carotenoids in diabetes. AB - OBJECTIVE: A case-control study was conducted to evaluate the effects of diabetes mellitus on serum levels of vitamin A, alpha-carotene, beta-carotene, alpha tocopherol, serum and urine RBP. SUBJECTS: One hundred and seven patients with Type 2 diabetes mellitus (28-74 y) were recruited from those attending a primary health care clinic in King Khalid University Hospital in Riyadh City (Saudi Arabia). They were matched for age and sex with 143 healthy individuals. METHODS: Fasting blood samples and 10h urine collections were obtained from all subjects. Levels of vitamins and carotenoids in serum measured by high performance liquid chromatography (HPLC), and of retinol binding protein (RBP) in serum and urine by an enzyme-linked immunosorbent assay (ELISA). RESULTS: The mean serum concentrations of retinol, alpha-carotene, and alpha-tocopherol were similar in both groups after correction of lipid soluble vitamins for serum lipids levels. However, serum beta-carotene concentration was significantly higher in control subjects than diabetics (P = 0.002). Serum and urine RBP concentrations were significantly higher in diabetics than in controls (P = 0.0001). In normal subjects (but not diabetics) serum concentrations of retinol and RBP were higher in men than in women (P = 0.02, P = 0.0001 respectively). In both normal and diabetic subjects, serum levels of alpha-tocopherol (P = 0.007) and urine RBP (P = 0.005), were higher in men than women. Urinary excretion of RBP was significantly higher in diabetic patients with renal impairment than other diabetics or controls (P = 0.0001). There was a negative correlation between fasting blood glucose (FBG) concentration and serum beta-carotene (P = 0.008) in the total combined group and a positive correlation between FBG and urinary RBP/creatinine (P = 0.009) in diabetic patients. CONCLUSION: Serum beta-carotene concentration was significantly lower in diabetic patients than controls. Serum retinol concentration in patients with diabetes was normal, yet serum and urine RBP concentrations were significantly higher in diabetics than in controls. PMID- 10477251 TI - Physical activity patterns of rural Senegalese adolescent girls during the dry and rainy seasons measured by movement registration and direct observation methods. AB - OBJECTIVE: To study the physical activity patterns and daily estimates of energy expenditure (EE) of rural adolescent Senegalese girls. DESIGN: Physical activity was monitored using a portable accelerometer during four consecutive days and within the same time, by direct observation during two consecutive days. Adolescent girls were followed during the dry season (June, n = 40) and again during the rainy season (September, n = 30) SETTING: The Niakhar district in the central part of Senegal. SUBJECTS: Forty adolescents (13.3+/-0.5 y) drawn from a sample of 221 rural girls followed as part of a longitudinal study on growth and nutrition during puberty. RESULTS: Reliability of movement counts was acceptable (intraclass correlation, R = 0.71). There was a linear relationship between movement counts and observed scores. Predicted physical activity levels were high: 1.90+/-0.12 Mets (EE: 9.03+/-0.77 MJ). Physical activity levels derived from movement counts during the day are higher in the rainy season than during the dry season. This coincided with a depression in nutritional indicators during the rainy season apparent in the overall sample. Sleep duration appeared to be short in both seasons (6-6.5h per night). Senegalese adolescents participated in daily household tasks but time spent in productive activities, agricultural or handicraft, was less than 1 h per day. CONCLUSIONS: Estimates of activity-related EE of Senegalese adolescents were greater than those of adolescents from developed countries but closed to recent estimates for rural areas in developing countries. Energy requirements drawn from developed countries do not necessarily apply to African adolescent girls. PMID- 10477252 TI - Carotenoids in young and elderly healthy humans: dietary intakes, biochemical status and diet-plasma relationships. AB - OBJECTIVE: To determine dietary carotenoid concentrations using an established and newly developed food frequency questionnaire (FFQ) method, to determine plasma carotenoid concentrations and to determine the relationship between these dietary and plasma variables in 24-45 y and > or = 65 y groups. DESIGN: Descriptive assessment of (FFQ), 7-d estimated records, and plasma carotenoids and their relationships in 24-45 y and > or = 65 y groups. SETTING: Free living urban adults in Ireland. SUBJECTS: Sixty-four volunteers aged 24-45 y and 54 volunteers aged > or = 65 y. RESULTS: Beta-carotene was the predominant plasma carotenoid, but older groups had lower plasma concentrations of several carotenoids compared to younger groups (P < 0.005). Beta-carotene and lycopene were the major dietary carotenoids reported by estimated records and FFQ. Several estimated record and plasma carotenoid concentrations were positively associated in younger groups but not in older groups. FFQ overestimated dietary carotenoids relative to estimated records (P < or = 0.05), generally did not reflect estimated record carotenoid concentrations and showed positive associations with plasma carotenoids only in older men. Neither of the dietary methods revealed a positive association between plasma and dietary beta-carotene concentrations, whereas beta-cryptoxanthin was strongly associated. CONCLUSIONS: Dietary and plasma concentrations of individual carotenoids are documented in young and elderly groups of a European country. Estimated record data reveals positive associations between diet and plasma carotenoids in younger, but not elderly groups. Further work examining diet-plasma relationship in older groups and developing a common FFQ suitable for use in several European countries is required. PMID- 10477253 TI - Nutrient intakes and cholesterol values of the parents in a prospective randomized child-targeted coronary heart disease risk factor intervention trial- the STRIP project. AB - OBJECTIVE: To analyze food consumption, nutrient intakes and serum cholesterol concentrations of the parents in a child-targeted CHD intervention trial, during which the age of children increased from 7 months to 5 y. DESIGN AND SUBJECTS: The children were randomized to an intervention group (n = 540) or a control group (n = 522) at six months of age. The intervention families were counseled at 3-6 month intervals to reduce their child's intake of saturated fat and cholesterol. Dietary issues were discussed with the control families only briefly. The parents' food consumption was analyzed by 24 h dietary recall at the child's age of 7 and 13 months and at 2, 3, 4, and 5 y. Nutrient intakes were calculated using the Micro-Nutrica program. RESULTS: The mothers and fathers of the intervention children used less butter, more margarine and more skim milk than those of the control children (P < 0.001 for all measurements). After the onset of counseling, the intervention mothers consumed continuously less fat (1.4 E% less at the child's age of 5 y), less saturated fat (1.5 E% less at the child's age of 5 y) and more polyunsaturated fat (0.5 E% more at the child's age of 5 y) than the control mothers (P = 0.008, P < 0.001 and P < 0.001 for trend, respectively). After the child's age of 13 months the intervention fathers also had a continuously lower fat intake (2.4 E% less at the child's age of 5 y) and consumed less saturated fat (1.5 E% less at the child's age of 5 y) than the control fathers (P < 0.001 for trend for both measurements). The serum cholesterol concentration of the intervention mothers was consistently lower than that of the control mothers during the intervention (at child's age of 5 y 4.86 and 5.09 mmol/L, respectively; P for trend = 0.03), while the values of the intervention and control fathers showed no differences. CONCLUSIONS: Continuous dietary intervention begun in infancy and focused on modification of the child's diet according to the current principles of preventive cardiology, was accompanied by a moderate decrease in the intake of total and saturated fat in the parents, but serum cholesterol concentration diminished consistently only in the mothers of the intervention children. PMID- 10477254 TI - Diet in relation to socioeconomic status in patients with coronary heart disease. AB - OBJECTIVES: Nutrient intake, food consumption and fatty acid profile of serum cholesteryl esters (CE) were investigated in relation to education in patients with coronary heart disease (CHD). DESIGN: A cross-sectional survey in CHD patients selected from hospital records at least six months after hospitalisation. Food record, a questionnaire on dietary fat and fatty acid profile of CE were used to examine diet composition. Years of education, divided into three categories, were used to indicate socioeconomic status. SETTING: The district of Kuopio University Hospital in Finland. SUBJECTS: One hundred and nine patients with coronary artery bypass surgery, 106 patients with coronary angioplasty, 101 patients with acute myocardial infarction and 99 patients with acute myocardial ischaemia aged 61 y (33-74 y) (mean (range)). RESULTS: Men with a low level of schooling had a higher intake of total and saturated fat, a lower consumption of vegetables and fruits, more frequently used butter or butter based spread and less frequently used oil compared to the diets of men with middle or high education. Men with low or middle education had a lower intake of alcohol than men with a high level of education. Highly-educated women had a lower proportion of myristic acid in CE than low-educated women and a similar trend between myristic acid and educational level was found in men. CONCLUSIONS: The educational level of men with CHD influenced their nutrient intake and food consumption, but in women with CHID its impact seemed to be weaker. PMID- 10477255 TI - Antioxidant status in children with protein-energy malnutrition (PEM) living in Cairo, Egypt. AB - OBJECTIVES: Free radicals are implicated in many diseases. The rise in free radicals associated with antioxidant deficiency results in tissue damage. The pathogenesis of oedema and anaemia commonly found in children with protein-energy malnutrition (PEM) has been suggested to be caused by an imbalance between the production of toxic radicals and their safe disposal. The aim of this study was to evaluate antioxidant status in children with PEM. DESIGN: A total of 68 children (age range: 3 months to 3 years) living in Cairo, Egypt were recruited. Forty-six of these subjects had different degrees of PEM; they were admitted at the Abo-Elrish Hospital, Cairo. Of these, 26 children had kwashiorkor (KWO) and twenty had marasmus (MAR). Twenty-two age and sex matched healthy well-nourished children were recruited from the local community, and used as controls. METHODS: The antioxidant status of the study population was determined by measuring copper zinc superoxide dismutase (Cu-Zn, SOD) in red blood cells, glutathione peroxidase (GPX) in whole blood, and ceruloplasmin in plasma. In addition, the plasma levels of trace-elements involving antioxidant activities, such as copper (Cu), zinc (Zn), and selenium (Se) were determined, along with a select group of vitamins. The latter included vitamin A (retinol), vitamin E (alpha-tocopherol) and vitamin C (ascorbic acid). RESULTS: The mean whole blood GPX activity along with plasma levels of vitamins A, E and C as well as ceruloplasmin, Cu and Se were all lower in children with either KWO or MAR than their corresponding control subjects. The erythrocyte SOD activity, on the other hand, was increased while the plasma Zn concentrations were either increased or not changed in the malnourished children. It was of interest that while haemoglobin concentrations were decreased, the plasma free iron (Fe) levels were significantly increased in children with KWO. CONCLUSIONS: The significant increase in red cell SOD activity associated with the decrease in plasma ceruloplasmin, antioxidant vitamins and the whole blood GPX activity in PEM children suggest that these children are potentially susceptible to high oxidative stress. An elevated plasma Fe concentrations, especially with KWO may augment the harmful effect of free radicals with a clinical consequence of oedema. PMID- 10477256 TI - Crystal structure of human mitochondrial NAD(P)+-dependent malic enzyme: a new class of oxidative decarboxylases. AB - BACKGROUND: Malic enzymes catalyze the oxidative decarboxylation of malate to pyruvate and CO2 with the concomitant reduction of NAD(P)+ to NAD(P)H. They are widely distributed in nature and have important biological functions. Human mitochondrial NAD(P)+-dependent malic enzyme (mNAD-ME) may have a crucial role in the metabolism of glutamine for energy production in rapidly dividing cells and tumors. Moreover, this isoform is unique among malic enzymes in that it is a cooperative enzyme, and its activity is controlled allosterically. RESULTS: The crystal structure of human mNAD-ME has been determined at 2.5 A resolution by the selenomethionyl multiwavelength anomalous diffraction method and refined to 2.1 A resolution. The structure of the monomer can be divided into four domains; the active site of the enzyme is located in a deep cleft at the interface between three of the domains. Three acidic residues (Glu255, Asp256 and Asp279) were identified as ligands for the divalent cation that is required for catalysis by malic enzymes. CONCLUSIONS: The structure reveals that malic enzymes belong to a new class of oxidative decarboxylases. The tetramer of the enzyme appears to be a dimer of dimers. The active site of each monomer is located far from the tetramer interface. The structure also shows the binding of a second NAD+ molecule in a pocket 35 A away from the active site. The natural ligand for this second binding site may be ATP, an allosteric inhibitor of the enzyme. PMID- 10477258 TI - High-efficiency Semliki Forest virus-mediated transduction in bovine adrenal chromaffin cells. AB - Adrenal chromaffin cells are commonly used in studies of exocytosis. Progress in characterizing the molecular mechanisms has been slow, because no simple, high efficiency technique is available for introducing and expressing heterologous cDNA in chromaffin cells. Here we demonstrate that Semliki Forest virus (SFV) vectors allow high-efficiency expression of heterologous protein in chromaffin cells. PMID- 10477259 TI - Salicylate-enhanced activation of transcription factors induced by interferon gamma. AB - Salicylate enhanced the interferon-gamma-dependent activation of two transcription factors in a murine macrophage cell line: signal transducer and activator of transcription (STAT)1 and interferon-gamma-responsive factor 1. Salicylate alone did not activate these transcription factors. This enhancement was reflected by increased DNA-binding activities and was the consequence of prolonged tyrosine phosphorylation of these transcription factors following interferon-gamma treatment. However, salicylate did not directly inhibit protein tyrosine phosphatase activity in nuclear extracts of interferon-gamma-treated cells. The enhanced activation of STAT1 resulted in increased induction of mRNA encoding interferon regulatory factor-1. These results not only demonstrate that aspirin and its metabolite salicylate may have pro-inflammatory as well as anti inflammatory effects but also raise the possibility that new cellular targets may be identified for modulating the Janus kinase-STAT signalling pathway. PMID- 10477260 TI - Ornithine and glutamate decarboxylases catalyse an oxidative deamination of their alpha-methyl substrates. AB - Ornithine decarboxylase (ODC) from Lactobacillus 30a catalyses the cleavage of alpha-methylornithine into ammonia and 2-methyl-1-pyrroline; glutamate decarboxylase (GAD) from Escherichia coli catalyses the cleavage of alpha methylglutamate into ammonia and laevulinic acid. In our analyses, 2-methyl-1 pyrroline and laevulinic acid were identified by HPLC and mass spectroscopic analysis, and ammonia was identified by means of glutamate dehydrogenase. Molecular oxygen was consumed during these reactions in a 1:2 molar ratio with respect to the products. The catalytic efficiencies (k(cat)/K(m)) of the reactions catalysed by ODC and GAD were determined as 12500 and 9163 M(-1).min( 1) respectively. When the reactions were performed under anaerobic conditions, no ammonia, 2-methyl-1-pyrroline or laevulinic acid was produced to a significant extent. The formation of ammonia and O(2) consumption (in a 1:2 molar ratio with respect to ammonia) were also detected during the reaction of ODC and GAD with putrescine and gamma-aminobutyrate respectively. Taken together, these findings clearly indicate that ODC and GAD catalyse an oxidative deamination of their decarboxylation products, a reaction similar to that catalysed by dopa decarboxylase (DDC) with alpha-methyldopa [Bertoldi, Dominici, Moore, Maras and Borri Voltattorni (1998) Biochemistry 37, 6552-6561]. Furthermore, this reaction was accompanied by a decarboxylation-dependent transamination occurring for GAD, DDC and ODC with a frequency of approx. 0.24%, 1% and 9% respectively compared with that of oxidative deamination. PMID- 10477257 TI - Repression of gene expression by oxidative stress. AB - Gene expression is modulated by both physiological signals (hormones, cytokines, etc.) and environmental stimuli (physical parameters, xenobiotics, etc.). Oxidative stress appears to be a key pleiotropic modulator which may be involved in either pathway. Indeed, reactive oxygen species (ROS) have been described as second messengers for several growth factors and cytokines, but have also been shown to rise following cellular insults such as xenobiotic metabolism or enzymic deficiency. Extensive studies on the induction of stress-response genes by oxidative stress have been reported. In contrast, owing to the historical focus on gene induction, less attention has been paid to gene repression by ROS. However, a growing number of studies have shown that moderate (i.e. non cytotoxic) oxidative stress specifically down-regulates the expression of various genes. In this review, we describe the alteration of several physiological functions resulting from oxidative-stress-mediated inhibition of gene transcription. We will then focus on the repressive oxidative modulation of various transcription factors elicited by ROS. PMID- 10477261 TI - Characterization of prenylated protein methyltransferase in Leishmania. AB - Prenylated protein methyltransferase, an enzyme involved in the post translational modification of many signalling proteins, has been characterized in a parasitic flagellated protozoan, Leishmania donovani. The activity of this enzyme was monitored by the methylation of an artificial substrate, an S prenylated cysteine analogue, with S-adenosyl-l-[methyl-(3)H]methionine as methyl donor. More than 85% of the methyltransferase activity was associated with membranes. The enzyme methylates N-acetyl-S-trans, trans-farnesyl-l-cysteine and N-acetyl-S-all-trans-geranylgeranyl-l-cysteine, but N-acetyl-S-trans, trans geranyl-l-cysteine only very weakly. In contrast with the enzyme from mammals, the leishmanial enzyme had a greater affinity for the farnesylated substrate than for the geranylgeranylated one. Activity in vitro was not modulated by cAMP, protein kinase C activator or guanosine 5'-[gamma-thio]triphosphate. An analysis of the endogenous substrates showed that the carboxymethylated proteins were also isoprenylated. The main carboxymethylated proteins have molecular masses of 95, 68, 55, 46, 34-23, 18 and less than 14 kDa. Treatment of cells with N-acetyl-S trans,trans-farnesyl-l-cysteine decreased the carboxymethylation level, whereas treatment with guanosine 5'-[gamma-thio]triphosphate increased the carboxymethylation of various proteins, particularly those of molecular masses 30 20 kDa. PMID- 10477262 TI - Cellular stress in xenopus kidney cells enhances the phosphorylation of eukaryotic translation initiation factor (eIF)4E and the association of eIF4F with poly(A)-binding protein. AB - Eukaryotic initiation factor (eIF) 4E binds to the 5'-cap structure of eukaryotic mRNA and has a central role in the control of cell proliferation. We have shown previously that the stimulation of cultured Xenopus kidney cells with serum resulted in the activation of protein synthesis, enhanced phosphorylation of eIF4E and increased binding of the adapter protein, eIF4G, and poly(A)-binding protein (PABP) to eIF4E to form the functional initiation factor complex, eIF4F/PABP. We now show that cellular stresses such as arsenite, anisomycin and heat shock also result in increased phosphorylation of eIF4E, eIF4F complex formation and the association of PABP with eIF4G, in conditions under which the rate of protein synthesis is severely inhibited. In contrast with reported effects on mammalian cells, the stress-induced increase in eIF4F complex formation occurs in the absence of changes in the association of eIF4E with its binding proteins 4E-BP1 or 4E-BP2. The stress-induced changes in eIF4E phosphorylation were totally abrogated by the p38 mitogen-activated protein (MAP) kinase inhibitor SB203580, and were partly inhibited by the phosphoinositide 3 kinase inhibitor LY294002 and the mammalian target of rapamycin (mTOR) inhibitor rapamycin. However, eIF4E phosphorylation was unaffected by extracellular signal regulated protein kinase (MAP kinase) inhibitor PD98059. These results indicate that cellular stresses activate multiple signalling pathways that converge at the level of eIF4F complex formation to influence the interactions between eIF4E, eIF4G and PABP. PMID- 10477263 TI - Structural determination of a 5-acetamido-3,5,7, 9-tetradeoxy-7-(3 hydroxybutyramido)-L-glycero-L-manno-nonulos onic acid-containing homopolysaccharide isolated from Sinorhizobium fredii HH103. AB - The structure of a polysaccharide from Sinorhizobium fredii HH103 has been determined. This polysaccharide was isolated by following the protocol for lipopolysaccharide extraction. On the basis of monosaccharide analysis, methylation analysis, fast atom bombardment MS, matrix-assisted laser desorption ionization MS, electron-impact high-resolution MS, one-dimensional (1)H-NMR and (13)C-NMR and two-dimensional NMR experiments, the structure was shown to consist of a homopolymer of a 3:1 mixture of 5-acetamido-3,5,7, 9-tetradeoxy-7-[(R)- and (S)-3-hydroxybutyramido]-l-glycero-l-manno-nonulosonic acid. The sugar residues are attached via a glycosidic linkage to the OH group of the 3-hydroxybutyramido substituent and thus the monomers are linked via both glycosidic and amidic linkages. In contrast with the Sinorhizobium K-antigens previously reported, which are composed of a disaccharide repeating unit, the K-antigen polysacharide of S. fredii HH103 is a homopolysaccharide. PMID- 10477264 TI - Defective kinetics of cytochrome c oxidase and alteration of mitochondrial membrane potential in fibroblasts and cytoplasmic hybrid cells with the mutation for myoclonus epilepsy with ragged-red fibres ('MERRF') at position 8344 nt. AB - We have investigated pathogenic effects of the tRNA(Lys) A8344G mutation associated with the syndrome myoclonus epilepsy with ragged-red fibres (MERRF) by using fibroblasts and fibroblast-derived cytoplasmic hybrid cells harbouring different percentages of mutated mitochondrial DNA (mtDNA). The activity of cytochrome c oxidase (COX) in patient fibroblasts with 89% mutated mtDNA was decreased to 20% of the control levels. COX exhibited altered kinetics, with a decreased V(max) for both the low-affinity and high-affinity phases; however, the K(m) values were not significantly changed. The substrate-dependent synthesis of ATP was decreased to 50% of the control. Analysis of the mitochondrial membrane potential, DeltaPsi, in digitonin-treated cells with tetramethylrhodamine methyl ester (TMRM) with the use of flow cytometry showed a 80% decrease in DeltaPsi at state 4 and an increased sensitivity of DeltaPsi to an uncoupler in fibroblasts from the patient. The investigation of transmitochondrial cytoplasmic hybrid clones derived from the patient's fibroblasts enabled us to characterize the relationship between heteroplasmy of the MERRF mutation, COX activity and DeltaPsi. Within the range of 87-73% mutated mtDNA, COX activity was decreased to 5-35% and DeltaPsi was decreased to 6-78%. These results demonstrate that the MERRF mutation affects COX activity and DeltaPsi in different proportions with regard to mutation heteroplasmy and indicate that the biochemical manifestation of the MERRF mutation exerts a very steep threshold of DeltaPsi inhibition. PMID- 10477265 TI - Inhibition of glycogenolysis in primary rat hepatocytes by 1, 4-dideoxy-1,4-imino D-arabinitol. AB - 1,4-Dideoxy-1,4-imino-d-arabinitol (DAB) was identified previously as a potent inhibitor of both the phosphorylated and non-phosphorylated forms of glycogen phosphorylase (EC 2.4.1.1). In the present study, the effects of DAB were investigated in primary cultured rat hepatocytes. The transport of DAB into hepatocytes was dependent on time and DAB concentration. The rate of DAB transport was 192 pmol/min per mg of protein per mM DAB(medium-concentration). In hepatocytes, DAB inhibited basal and glucagon-stimulated glycogenolysis with IC(50) values of 1.0+/-0.3 and 1.1+/-0.2 microM, respectively. The primary inhibitory effect of DAB on glycogenolysis was shown to be due to inhibition of glycogen phosphorylase but, at higher concentrations of DAB, inhibition of the debranching enzyme (4-alpha-glucanotransferase, EC 2.4.1.25) may have an effect. No effects on glycogen synthesis were observed, demonstrating that glycogen recycling does not occur in cultured hepatocytes under the conditions tested. Furthermore, DAB had no effects on phosphorylase kinase, the enzyme responsible for phosphorylation and thereby activation of glycogen phosphorylase, or on protein phosphatase 1, the enzyme responsible for inactivation of glycogen phosphorylase through dephosphorylation. PMID- 10477266 TI - Effects of disrupting the 21 kDa subunit of complex I from Neurospora crassa. AB - We have cloned and inactivated in vivo, by repeat-induced point mutations, the nuclear gene encoding a 21 kDa subunit of complex I from Neurospora crassa. Mitochondria from the nuo21 mutant lack this specific protein but retain other subunits of complex I in approximately normal amounts. In addition, this mutant is able to assemble an almost intact enzyme. The electron transfer activities from NADH to artificial acceptors of mitochondrial membranes from nuo21 differ from those of the wild-type strain, suggesting that the absence of the 21 kDa polypeptide results in conformational changes in complex I. Nevertheless, complex I of nuo21 is able to perform NADH:ubiquinone reductase activity, as judged by the observation that the respiration of mutant mitochondria is sensitive to inhibition by rotenone. We discuss these findings in relation to the involvement of complex I in mitochondrial diseases. PMID- 10477267 TI - Acetate stimulates flux through the tricarboxylic acid cycle in rabbit renal proximal tubules synthesizing glutamine from alanine: a 13C NMR study. AB - Although glutamine synthesis has a major role in the control of acid-base balance and ammonia detoxification in the kidney of herbivorous species, very little is known about the regulation of this process. We therefore studied the influence of acetate, which is readily metabolized by the kidney and whose metabolism is accompanied by the production of bicarbonate, on glutamine synthesis from variously labelled [(13)C]alanine and [(14)C]alanine molecules in isolated rabbit renal proximal tubules. With alanine as sole exogenous substrate, glutamine and, to a smaller extent, glutamate and CO(2), were the only significant products of the metabolism of this amino acid, which was removed at high rates. Absolute fluxes through the enzymes involved in alanine conversion into glutamine were assessed by using a novel model describing the corresponding reactions in conjunction with the (13)C NMR, and to a smaller extent, the radioactive and enzymic data. The presence of acetate (5 mM) led to a large stimulation of fluxes through citrate synthase and alpha-oxoglutarate dehydrogenase. These effects were accompanied by increases in the removal of alanine, in the accumulation of glutamate and in flux through the anaplerotic enzyme pyruvate carboxylase. Acetate did not alter fluxes through glutamate dehydrogenase and glutamine synthetase; as a result, acetate did not change the accumulation of ammonia, which was negligible under both experimental conditions. We conclude that acetate, which seems to be an important energy-provider to the rabbit renal proximal tubule, simultaneously traps as glutamate the extra nitrogen removed as alanine, thus preventing the release of additional ammonia by the glutamate dehydrogenase reaction. PMID- 10477268 TI - Model of 2,3-bisphosphoglycerate metabolism in the human erythrocyte based on detailed enzyme kinetic equations: in vivo kinetic characterization of 2,3 bisphosphoglycerate synthase/phosphatase using 13C and 31P NMR. AB - This is the first in a series of three papers [see also Mulquiney and Kuchel (1999) Biochem. J. 342, 579-594; Mulquiney and Kuchel (1999) Biochem. J. 342, 595 602] that present a detailed mathematical model of erythrocyte metabolism which explains the regulation and control of 2,3-bisphosphoglycerate (2,3-BPG) metabolism. 2,3-BPG is a modulator of haemoglobin oxygen affinity and hence plays an important role in blood oxygen transport and delivery. This paper presents an in vivo kinetic characterization of 2,3-BPG synthase/phosphatase (BPGS/P), the enzyme that catalyses both the synthesis and degradation of 2,3-BPG. Much previous work had indicated that the behaviour of this enzyme in vitro is markedly different from that in vivo. (13)C and (31)P NMR were used to monitor the time courses of selected metabolites when erythrocytes were incubated with or without [U-(13)C]glucose. Simulations of the experimental time courses were then made. By iteratively changing the parameters of the BPGS/P part of the model until a good match between the NMR-derived data and simulations were achieved, it was possible to characterize BPGS/P kinetically in vivo. This work revealed that: (1) the pH-dependence of the synthase activity results largely from a strong co operative inhibition of the synthase activity by protons; (2) 3-phosphoglycerate and 2-phosphoglycerate are much weaker inhibitors of 2,3-BPG phosphatase in vivo than in vitro; (3) the K(m) of BPGS/P for 2,3-BPG is significantly higher than that measured in vitro; (4) the maximal activity of the phosphatase in vivo is approximately twice that in vitro, when P(i) is the sole activator (second substrate); and (5) 2-phosphoglycollate appears to play no role in the activation of the phosphatase in vivo. Using the newly determined kinetic parameters, the percentage of glycolytic carbon flux that passes through the 2, 3-BPG shunt in the normal in vivo steady state was estimated to be 19%. PMID- 10477270 TI - Model of 2,3-bisphosphoglycerate metabolism in the human erythrocyte based on detailed enzyme kinetic equations: computer simulation and metabolic control analysis. AB - This is the third of three papers [see also Mulquiney, Bubb and Kuchel (1999) Biochem. J. 342, 565-578; Mulquiney and Kuchel (1999) Biochem. J. 342, 579-594] for which the general goal was to explain the regulation and control of 2,3 bisphosphoglycerate (2,3-BPG) metabolism in human erythrocytes. 2,3-BPG is a major modulator of haemoglobin oxygen affinity and hence is vital in blood oxygen transport. A detailed mathematical model of erythrocyte metabolism was presented in the first two papers. The model was refined through an iterative loop of experiment and simulation and it was used to predict outcomes that are consistent with the metabolic behaviour of the erythrocyte under a wide variety of experimental and physiological conditions. For the present paper, the model was examined using computer simulation and Metabolic Control Analysis. The analysis yielded several new insights into the regulation and control of 2,3-BPG metabolism. Specifically it was found that: (1) the feedback inhibition of hexokinase and phosphofructokinase by 2, 3-BPG are equally as important as the product inhibition of 2,3-BPG synthase in controlling the normal in vivo steady state concentration of 2,3-BPG; (2) H(+) and oxygen are effective regulators of 2,3-BPG concentration and that increases in 2,3-BPG concentrations are achieved with only small changes in glycolytic rate; (3) these two effectors exert most of their influence through hexokinase and phosphofructokinase; (4) flux through the 2,3-BPG shunt changes in absolute terms in response to different energy demands placed on the cell. This response of the 2,3-BPG shunt contributes an [ATP] stabilizing effect. A 'cost' of this is that 2, 3-BPG concentrations are very sensitive to the energy demand of the cell and; (5) the flux through the 2,3-BPG shunt does not change in response to different non-glycolytic demands for NADH. PMID- 10477269 TI - Model of 2,3-bisphosphoglycerate metabolism in the human erythrocyte based on detailed enzyme kinetic equations: equations and parameter refinement. AB - Over the last 25 years, several mathematical models of erythrocyte metabolism have been developed. Although these models have identified the key features in the regulation and control of erythrocyte metabolism, many important aspects remain unexplained. In particular, none of these models have satisfactorily accounted for 2,3-bisphosphoglycerate (2,3-BPG) metabolism. 2,3-BPG is an important modulator of haemoglobin oxygen affinity, and hence an understanding of the regulation of 2,3-BPG concentration is important for understanding blood oxygen transport. A detailed, comprehensive, and hence realistic mathematical model of erythrocyte metabolism is presented that can explain the regulation and control of 2,3-BPG concentration and turnover. The model is restricted to the core metabolic pathways, namely glycolysis, the 2,3-BPG shunt and the pentose phosphate pathway (PPP), and includes membrane transport of metabolites, the binding of metabolites to haemoglobin and Mg(2+), as well as pH effects on key enzymic reactions and binding processes. The model is necessarily complex, since it is intended to describe the regulation and control of 2,3-BPG metabolism under a wide variety of physiological and experimental conditions. In addition, since H(+) and blood oxygen tension are important external effectors of 2,3-BPG concentration, it was important that the model take into account the large array of kinetic and binding phenomena that result from changes in these effectors. Through an iterative loop of experimental and simulation analysis many values of enzyme-kinetic parameters of the model were refined to yield close conformity between model simulations and 'real' experimental data. This iterative process enabled a single set of parameters to be found which described well the metabolic behaviour of the erythrocyte under a wide variety of conditions. PMID- 10477271 TI - Characterization and polyanion-binding properties of purified recombinant prion protein. AB - Certain polysulphated polyanions have been shown to have prophylactic effects on the progression of transmissible spongiform encephalopathy disease, presumably because they bind to prion protein (PrP). Until now, the difficulty of obtaining large quantities of native PrP has precluded detailed studies of these interactions. We have over-expressed murine recombinant PrP (recPrP), lacking its glycophosphoinositol membrane anchor, in modified mammalian cells. Milligram quantities of secreted, soluble and partially glycosylated protein were purified under non-denaturing conditions and the identities of mature-length aglycosyl recPrP and two cleavage fragments were determined by electrospray MS. Binding was assessed by surface plasmon resonance techniques using both direct and competitive ligand-binding approaches. recPrP binding to immobilized polyanions was enhanced by divalent metal ions. Polyanion binding was strong and showed complex association and dissociation kinetics that were consistent with ligand directed recPrP aggregation. The differences in the binding strengths of recPrP to pentosan polysulphate and to other sulphated polyanions were found to parallel their in vivo anti-scrapie and in vitro anti-scrapie-specific PrP formation potencies. When recPrP was immobilized by capture on metal-ion chelates it was found, contrary to expectation, that the addition of polyanions promoted the dissociation of the protein. PMID- 10477272 TI - Signal transduction by beta1 integrin receptors in human chondrocytes in vitro: collaboration with the insulin-like growth factor-I receptor. AB - We have examined the mechanism by which collagen-binding integrins co-operate with insulin-like growth factor-I (IGF-I) receptors (IGF-IR) to regulate chondrocyte phenotype and differentiation. Adhesion of chondrocytes to anti-beta1 integrin antibodies or collagen type II leads to phosphorylation of cytoskeletal and signalling proteins localized at focal adhesions, including alpha-actinin, vinculin, paxillin and focal adhesion kinase (FAK). These stimulate docking proteins such as Shc (Src-homology collagen). Moreover, exposure of collagen type II-cultured chondrocytes to IGF-I leads to co-immunoprecipitation of Shc protein with the IGF-IR and with beta1, alpha1 and alpha5 integrins, but not with alpha3 integrin. Shc then associates with growth factor receptor-bound protein 2 (Grb2), an adaptor protein and extracellular signal-regulated kinase. The expression of the docking protein Shc occurs only when chondrocytes are bound to collagen type II or integrin antibodies and increases when IGF-I is added, suggesting a collaboration between integrins and growth factors in a common/shared biochemical signalling pathway. Furthermore, these results indicate that focal adhesion assembly may facilitate signalling via Shc, a potential common target for signal integration between integrin and growth-factor signalling regulatory pathways. Thus, the collagen-binding integrins and IGF-IR co-operate to regulate focal adhesion components and these signalling pathways have common targets (Shc-Grb2 complex) in subcellular compartments, thereby linking to the Ras-mitogen activated protein kinase signalling pathway. These events may play a role during chondrocyte differentiation. PMID- 10477273 TI - Production and functional activity of a recombinant von Willebrand factor-A domain from human complement factor B. AB - Factor B is a five-domain 90 kDa serine protease proenzyme which is part of the human serum complement system. It binds to other complement proteins C3b and properdin, and is activated by the protease factor D. The fourth domain of factor B is homologous to the type A domain of von Willebrand Factor (vWF-A). A full length human factor B cDNA clone was used to amplify the region encoding the vWF A domain (amino acids 229-444 of factor B). A fusion protein expression system was then used to generate it in high yield in Escherichia coli, where thrombin cleavage was used to separate the vWF-A domain from its fusion protein partner. A second vWF-A domain with improved stability and solubility was created using a Cys(267)-->Ser mutation and a four-residue C-terminal extension of the first vWF A domain. The recombinant domains were investigated by analytical gel filtration, sucrose density centrifugation and analytical ultracentrifugation, in order to show that both domains were monomeric and possessed compact structures that were consistent with known vWF-A crystal structures. This expression system and its characterization permitted the first investigation of the function of the isolated vWF-A domain. It was able to inhibit substantially the binding of (125)I labelled factor B to immobilized C3b. This demonstrated both the presence of a C3b binding site in this portion of factor B and a ligand-binding property of the vWF-A domain. The site at which factor D cleaves factor B is close to the N terminus of both recombinant vWF-A domains. Factor D was shown to cleave the vWF A domain in the presence or absence of C3b, whereas the cleavage of intact factor B under the same conditions occurs only in the presence of C3b. PMID- 10477274 TI - GA2/GM2/GD2 synthase localizes to the trans-golgi network of CHO-K1 cells. AB - UDP-GalNAc:lactosylceramide/GM3/GD3 beta-1,4-N-acetylgalactosaminyltransferase (GalNAc-T) transforms its acceptors into the gangliosides GA2, GM2 and GD2. It is well established that it is a Golgi-located glycosyltransferase, but its sub Golgi localization is still unclear. We addressed this question in Chinese hamster ovary K1 cell clones stably transfected with a c-myc-tagged version of GalNAc-T which express the enzyme at different levels of activity. In these cell clones we examined the effect of brefeldin A (BFA) on the synthesis of glycolipids (in metabolic-labelling experiments) and on the sub-Golgi localization of the GalNAc-T (by immunocytochemistry). We found that in cell clones expressing moderate levels of activity, GalNAc-T immunoreactivity behaved as the trans-Golgi network (TGN) marker mannose-6-P receptor (M6PR) both in BFA treated and untreated cells, and that BFA completely blocked the synthesis of GM2, GM1 and GD1a. On the other hand, in cell clones expressing high levels of activity and treated with BFA, most GalNAc-T immunoreactivity redistributed to the endoplasmic reticulum, as did the medial-Golgi marker mannosidase II, and the synthesis of GM2, GM1 and GD1a was not completely blocked. These results indicate that GalNAc-T is a TGN-located enzyme and that the mechanism that localizes it to this compartment involves steps that, when saturated, lead to its mislocalization to the cis-, medial- or trans-Golgi. Changes of Golgi membrane properties by modification of local glycolipid composition due to the activity of the expressed enzyme were not the main cause of mislocalization, since it persists when glycolipid synthesis is inhibited with d, l-threo-1-phenyl-2-hexadecanoylamino-3 pyrrolidino-1-propanol-HCl. PMID- 10477275 TI - Localization of a carboxylic residue possibly involved in the inhibition of vacuolar H+-pyrophosphatase by N, N'-dicyclohexylcarbodi-imide. AB - A vacuolar H(+)-pyrophosphatase (EC 3.6.1.1) that catalyses PP(i) hydrolysis and the electrogenic translocation of protons from the cytosol to the vacuole lumen, was purified from etiolated hypocotyls of mung bean seedlings (Vigna radiata L.). Group-specific modification was used to identify a carboxylic residue involved in the inhibition of vacuolar H(+)-pyrophosphatase. Carbodi-imides, such as N,N' dicyclohexylcarbodi-imide (DCCD) and 1-ethyl-3-(3-dimethylamino-propyl)carbodi imide, and Woodward's reagent K caused a progressive decline in the enzymic activity of vacuolar H(+)-pyrophosphatase in a time- and concentration-dependent manner. The stoichiometry of labelling of the vacuolar H(+)-pyrophosphatase by [(14)C]DCCD determined that DCCD modifies one carboxylic residue per subunit of the enzyme. Protection studies suggest that the DCCD-reactive carboxylic residue resides at or near the substrate-binding site. Furthermore, peptide mapping analysis reveals that Asp(283), located in the putative loop V of a tentative topological model of vacuolar H(+)-pyrophosphatase on the cytosolic side, was labelled by radioactive [(14)C]DCCD. Cytosolic loop V contains both DCCD sensitive Asp(283) and a conserved motif sequence, rendering it a candidate for the catalytic site of vacuolar H(+)-pyrophosphatase. A topological picture of the active domain of vacuolar H(+)-pyrophosphatase is tentatively proposed. PMID- 10477276 TI - Interaction with amylopectin influences the ability of granule-bound starch synthase I to elongate malto-oligosaccharides. AB - This paper examines the properties in soluble form of two isoforms of starch synthase. One of these, granule-bound starch synthase I (GBSSI), is responsible for the synthesis of amylose inside the amylopectin matrix of the starch granule in vivo. The other, starch synthase II (SSII), is involved in amylopectin synthesis. Both isoforms can use amylopectin and malto-oligosaccharide as substrates in vitro. As well as acting as a substrate for GBSSI, amylopectin acts as an effector of this isoform, increasing the rate at which it elongates malto oligosaccharides and promoting a processive rather than distributive mode of elongation of these compounds. The affinity of GBSSI for amylopectin as an effector is greater than its affinity for amylopectin as a substrate. The rate and mode of elongation of malto-oligosaccharides by SSII are not influenced by amylopectin. These results suggest that specific interaction with amylopectin in the matrix of the starch granule is a unique property of GBSSI and is critical in determining the nature of its products. PMID- 10477277 TI - Caspase-1 (interleukin-1beta-converting enzyme) is inhibited by the human serpin analogue proteinase inhibitor 9. AB - The regulation of caspases, cysteine proteinases that cleave their substrates after aspartic residues, is poorly understood, even though they are involved in tightly regulated cellular processes. The recently discovered serpin analogue proteinase inhibitor 9 (PI9) is unique among human serpin analogues in that it has an acidic residue in the putative specificity-determining position of the reactive-site loop. We measured the ability of PI9 to inhibit the amidolytic activity of several caspases. The hydrolysis of peptide substrates by caspase-1 (interleukin-1beta-converting enzyme), caspase-4 and caspase-8 is inhibited by PI9 in a time-dependent manner. The rate of reaction of caspase-1 with PI9, as well as the rate of substrate hydrolysis of the initial caspase-PI9 complex, shows a hyperbolic dependence on the concentration of PI9, indicative of a two step kinetic mechanism for inhibition with an apparent second-order rate constant of 7x10(2) M(-1).s(-1). The hydrolysis of a tetrapeptide substrate by caspase-3 is not inhibited by PI9. The complexes of caspase-1 and caspase-4 with PI9 can be immunoprecipitated but no complex with caspase-3 can be detected. No complex can be immunoprecipitated if the active site of the caspase is blocked with a covalent inhibitor. These results show that PI9 is an inhibitor of caspase-1 and to a smaller extent caspase-4 and caspase-8, but not of the more distantly related caspase-3. PI9 is the first example of a human serpin analogue that inhibits members of this class of cysteine proteinases. PMID- 10477278 TI - The dihydrosphingosine-1-phosphate phosphatases of Saccharomyces cerevisiae are important regulators of cell proliferation and heat stress responses. AB - We have identified YSR2 and YSR3 of Saccharomyces cerevisiae as genes encoding dihydrosphingosine-1-phosphate phophatases which are involved in regulation of sphingolipid metabolism [Mao, Wadleigh, Jenkins, Hannun and Obeid (1997) J. Biol. Chem. 272, 28690-28694]. In this study, we explored the physiological roles that these enzymes may have in S. cerevisiae. Deletion of either YSR2, YSR3 or both did not affect viability or growth rate of yeast cells. However, overexpression of YSR2 significantly prolonged the doubling time of cell growth, whereas overexpression of YSR3 affected cell growth only slightly. Cell cycle analysis suggested that overexpression of either YSR2 or, to a lesser extent, YSR3 caused cell cycle arrest at the G1 phase. Disruption of YSR2, but not YSR3, conferred increased thermotolerance. On the other hand, overexpression of either YSR2 or YSR3 diminished thermotolerance. Using labelled dihydrosphingosine and dihydrosphingosine-1-P (DHS-1-P), we found that overexpression of YSR2 significantly increased ceramide formation, whereas deletion of YSR2, YSR3, or both, accumulated DHS-1-P, and deletion of YSR2 decreased ceramide formation. Together, these results show that the phenotypes of YSR2 are associated with changes in endogenous levels of the different sphingolipids. Green fluorescent protein tagging showed that in the exponentially growing cells, YSR2 and YSR3 had the same cellular localization to endoplasmic reticulum. However, YSR2 and YSR3 differ in mRNA levels: YSR2 had significantly higher mRNA levels than YSR3. This discrepancy might result in the functional differences that these proteins exhibited. In addition, this study implicates sphingolipids and their metabolism in the regulation of growth and heat stress responses of the yeast S. cerevisiae. PMID- 10477279 TI - Differential effects of fibroblast growth factor (FGF) 9 and FGF2 on proliferation, differentiation and terminal differentiation of chondrocytic cells in vitro. AB - Fibroblast growth factor (FGF) 9 was compared with FGF2 in its ability to influence proliferation, differentiation, terminal differentiation and apoptosis in a rat calvaria-derived cell line (RCJ 3.1C5.18) that spontaneously undergoes chondrocyte differentiation in vitro. Like FGF2, FGF9 promoted proliferation, but to a lesser extent. In contrast to FGF2, which blocked chondrocytic differentiation, FGF9 had no effect on differentiation but inhibited terminal differentiation. FGF9 also stimulated expression of the mitotic inhibitor p21 to a greater extent than FGF2. Neither ligand influenced apoptosis. The results indicate that FGF9 could account for many of the physiological responses attributed to FGF-receptor activation in the growth plate. PMID- 10477280 TI - Expression of translationally controlled tumour protein is regulated by calcium at both the transcriptional and post-transcriptional level. AB - We have investigated how the programme of protein synthesis is altered in response to a loss of calcium homoeostasis in Cos-7 cells using a differential proteome mapping approach. Exposure of the cells to the calcium ionophore A23187 or thapsigargin, or alternatively, expression of a viral glycoprotein reported to deplete intracellular calcium stores, resulted in the up-regulated expression of a characteristic set of proteins. One of these is the translationally controlled tumour protein (TCTP), a cytoplasmic protein whose expression has not previously been linked to calcium perturbation. Quantitative Northern blot assay demonstrated that steady-state mRNA abundance of TCTP was also increased under these conditions. Clamping the cytosolic calcium concentration by the introduction of bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetra-acetic acid (BAPTA) into cells did not affect the increase in steady-state levels of TCTP mRNA observed in response to ionophore. Therefore depletion of endoplasmic reticulum (ER) calcium, but not elevation of the cytosolic calcium concentration, was responsible for increased transcription of the TCTP gene. However, the presence of BAPTA significantly attenuated the ionophore-mediated increase in levels of the protein. Moreover, the level of TCTP in ionophore-treated cells increased in advance of a detectable increase in the corresponding mRNA abundance. These results indicate that expression of TCTP is regulated at two distinct levels in response to the concentration of calcium in different cellular compartments. Whereas depletion of the ER store causes an increase in TCTP mRNA abundance, increased cytosolic calcium concentrations regulate gene expression at the post transcriptional level. PMID- 10477281 TI - Iron-dependent regulation of transferrin receptor expression in Trypanosoma brucei. AB - Transferrin is an essential growth factor for African trypanosomes. Here we show that expression of the trypanosomal transferrin receptor, which bears no structural similarity with mammalian transferrin receptors, is regulated by iron availability. Iron depletion of bloodstream forms of Trypanosoma brucei with the iron chelator deferoxamine resulted in a 3-fold up-regulation of the transferrin receptor and a 3-fold increase of the transferrin uptake rate. The abundance of expression site associated gene product 6 (ESAG6) mRNA, which encodes one of the two subunits of the trypanosome transferrin receptor, is regulated 5-fold by a post-transcriptional mechanism. In mammalian cells the stability of transferrin receptor mRNA is controlled by iron regulatory proteins (IRPs) binding to iron responsive elements (IREs) in the 3'-untranslated region (UTR). Therefore, the role of a T. brucei cytoplasmic aconitase (TbACO) that is highly related to mammalian IRP-1 was investigated. Iron regulation of the transferrin receptor was found to be unaffected in Deltaaco::NEO/Deltaaco::HYG null mutants generated by targeted disruption of the TbACO gene. Thus, the mechanism of post transcriptional transferrin receptor regulation in trypanosomes appears to be distinct from the IRE/IRP paradigm. The transferrin uptake rate was also increased when trypanosomes were transferred from medium supplemented with foetal bovine serum to medium supplemented with sera from other vertebrates. Due to varying binding affinities of the trypanosomal transferrin receptor for transferrins of different species, serum change can result in iron starvation. Thus, regulation of transferrin receptor expression may be a fast compensatory mechanism upon transmission of the parasite to a new host species. PMID- 10477282 TI - Distribution of the src-homology-2-domain-containing inositol 5-phosphatase SHIP 2 in both non-haemopoietic and haemopoietic cells and possible involvement of SHIP-2 in negative signalling of B-cells. AB - The termination of activation signals is a critical step in the control of the immune response; perturbation of inhibitory feedback pathways results in profound immune defects culminating in autoimmunity and overwhelming inflammation. FcgammaRIIB receptor is a well described inhibitory receptor. The ligation of B cell receptor (BCR) and FcgammaRIIB leads to the inhibition of B-cell activation. Numerous studies have demonstrated that the SH2-domain-containing inositol 5 phosphatase SHIP (referred hereto as SHIP-1) is essential in this process. The cDNA encoding a second SH2-domain-containing inositol 5-phosphatase, SHIP-2, has been cloned [Pesesse, Deleu, De Smedt, Drayer and Erneux (1997) Biochem. Biophys. Res. Commun. 239, 697-700]. Here we report the distribution of SHIP-2 in mouse tissues: a Western blot analysis of mouse tissues reveals that SHIP-2 is expressed in both haemopoietic and non-haemopoietic cells. In addition to T-cell and B-cell lines, spleen, thymus and lung are shown to coexpress SHIP-1 and SHIP 2. Moreover, SHIP-2 is detected in fibroblasts, heart and different brain areas. SHIP-2 shows a maximal tyrosine phosphorylation and association to Shc after ligation of BCR to FcgammaRIIB but not after stimulation of BCR alone. Our results therefore suggest a possible role for SHIP-2 in the negative regulation of immunocompetent cells. PMID- 10477283 TI - nSec-1 (munc-18) interacts with both primed and unprimed syntaxin 1A and associates in a dimeric complex on adrenal chromaffin granules. AB - The target-SNARE syntaxin 1A is an essential component of the core machinery required for regulated exocytosis (where SNARE is the soluble N-ethylmaleimide sensitive fusion protein-attachment protein receptor). Syntaxin 1A interacts with a variety of other proteins, two of which, N-ethylmaleimide-sensitive fusion protein (NSF) and alpha-soluble NSF attachment protein (alpha-SNAP) have been suggested to impart a conformational rearrangement on this protein during a reaction referred to as priming. We have studied the effect of the primed state on the binding properties of syntaxin 1A and we have confirmed that primed syntaxin 1A no longer associated with alpha-SNAP or its cognate vesicle-SNARE, vesicle-associated membrane protein (VAMP). Under such conditions, however, it retained the ability to bind to nSec-1. It has been demonstrated that nSec-1, a regulatory protein also involved in neuronal exocytosis, binds syntaxin 1A with high affinity in vitro, although evidence for this physical interaction occurring in vivo has proven elusive. We analysed the subcellular distribution of these two proteins in fractions from bovine adrenal medulla and detected syntaxin 1A and nSec-1 in both plasma membrane and chromaffin-granule fractions. Using a cross linking approach with chromaffin-granule membranes we detected a putative dimeric complex composed of approx. 54% total granule membrane nSec-1 and approx. 30% total syntaxin 1A. The results of this study therefore suggest the possibility of nSec-1 interactions with primed syntaxin 1A and demonstrate a potentially significant interaction of syntaxin 1A and nSec-1 on the membranes of chromaffin granules. PMID- 10477284 TI - Editing of non-cognate aminoacyl adenylates by peptide synthetases. AB - Non-ribosomally formed peptides display both highly conserved and variable amino acid positions, the variations leading to a wide range of peptide families. Activation of the amino acid substrate proceeds in analogy to the ribosomal biosynthetic mechanism generating aminoacyl adenylate and acyl intermediates. To approach the mechanism of fidelity of amino acid selection, the stability of the aminoacyl adenylates was studied by employing a continuous coupled spectrophotometric assay. The apo-form of tyrocidine synthetase 1 (apo-TY1) was used, generating an l-phenylalanyl-adenylate intermediate stabilized by the interaction of two structural subdomains of the adenylation domain. Adenylates of substrate analogues have shown variable and reduced degrees of stability, thus leading to an enhanced generation of pyrophosphate due to hydrolysis and continuous adenylate formation. Discrimination of the non-aromatic amino acids l Leu and l-Met, or l-Phe analogues such as p-amino- and p-chloro-l-Phe derivatives, as well as the stereospecific selection of l-Phe, is supported by less-stable adenylate intermediates exhibiting elevated susceptibility to hydrolysis. Breakdown of the l-phenylalanyl intermediate utilizing 2'-deoxy-ATP as the nucleotide substrate was significantly enhanced compared with the natural analogue. Apo-TY1 engineered at positions involved in adenylate formation showed variable protection against hydrolysis. The results imply that stability of the aminoacyl intermediates may act as an essential factor in substrate selection and fidelity of non-ribosomal-peptide-forming systems. PMID- 10477285 TI - Cloning and sequencing of the cDNA species for mammalian dimeric dihydrodiol dehydrogenases. AB - Cynomolgus and Japanese monkey kidneys, dog and pig livers and rabbit lens contain dimeric dihydrodiol dehydrogenase (EC 1.3.1.20) associated with high carbonyl reductase activity. Here we have isolated cDNA species for the dimeric enzymes by reverse transcriptase-PCR from human intestine in addition to the above five animal tissues. The amino acid sequences deduced from the monkey, pig and dog cDNA species perfectly matched the partial sequences of peptides digested from the respective enzymes of these animal tissues, and active recombinant proteins were expressed in a bacterial system from the monkey and human cDNA species. Northern blot analysis revealed the existence of a single 1.3 kb mRNA species for the enzyme in these animal tissues. The human enzyme shared 94%, 85%, 84% and 82% amino acid identity with the enzymes of the two monkey strains (their sequences were identical), the dog, the pig and the rabbit respectively. The sequences of the primate enzymes consisted of 335 amino acid residues and lacked one amino acid compared with the other animal enzymes. In contrast with previous reports that other types of dihydrodiol dehydrogenase, carbonyl reductases and enzymes with either activity belong to the aldo-keto reductase family or the short-chain dehydrogenase/reductase family, dimeric dihydrodiol dehydrogenase showed no sequence similarity with the members of the two protein families. The dimeric enzyme aligned with low degrees of identity (14-25%) with several prokaryotic proteins, in which 47 residues are strictly or highly conserved. Thus dimeric dihydrodiol dehydrogenase has a primary structure distinct from the previously known mammalian enzymes and is suggested to constitute a novel protein family with the prokaryotic proteins. PMID- 10477287 TI - Building the heart piece by piece: modularity of cis-elements regulating Nkx2-5 transcription. AB - Heart formation in Drosophila is dependent on the homeobox gene tinman. The homeobox gene Nkx2-5 is closely related to tinman and is the earliest known marker for cardiogenesis in vertebrate embryos. Recent studies of cis-regulatory elements required for Nkx2-5 expression in the developing mouse heart have revealed an extraordinary array of independent cardiac enhancers, and associated negative regulatory elements, that direct transcription in distinct regions of the embryonic heart. These studies demonstrate the modularity in cardiac transcription, in which different regulatory elements respond to distinct sets of transcription factors to control gene expression in different compartments of the developing heart. We consider the potential mechanisms underlying such transcriptional complexity, its possible significance for cardiac function, and the implications for evolution of the multichambered heart. PMID- 10477286 TI - Calcium-dependent properties of CIB binding to the integrin alphaIIb cytoplasmic domain and translocation to the platelet cytoskeleton. AB - The alphaIIbbeta3 integrin receives signals in agonist-activated platelets, resulting in its conversion to an active conformation that binds fibrinogen, thereby mediating platelet aggregation. Fibrinogen binding to alphaIIbbeta3 subsequently induces a cascade of intracellular signalling events. The molecular mechanisms of this bi-directional alphaIIbbeta3-mediated signalling are unknown but may involve the binding of proteins to the integrin cytoplasmic domains. We reported previously the sequence of a novel 22-kDa, EF-hand-containing, protein termed CIB (calcium- and integrin-binding protein) that interacts specifically with the alphaIIb cytoplasmic domain in the yeast two-hybrid system. Further analysis of numerous tissues and cell lines indicated that CIB mRNA and protein are widely expressed. In addition, isothermal titration calorimetry indicated that CIB binds to an alphaIIb cytoplasmic-domain peptide in a Ca(2+)-dependent manner, with moderate affinity (K(d), 700 nM) and 1:1 stoichiometry. In aggregated platelets, endogenous CIB and alphaIIbbeta3 translocate to the Triton X-100-insoluble cytoskeleton in a parallel manner, demonstrating that the cellular localization of CIB is regulated, potentially by alphaIIbbeta3. Thus CIB may contribute to integrin-related functions by mechanisms involving Ca(2+) modulated binding to the alphaIIb cytoplasmic domain and changes in intracellular distribution. PMID- 10477288 TI - Bix4 is activated directly by VegT and mediates endoderm formation in Xenopus development. AB - The maternal T-box gene VegT, whose transcripts are restricted to the vegetal hemisphere of the Xenopus embryo, plays an essential role in early development. Depletion of maternal VegT transcripts causes embryos to develop with no endoderm, while vegetal blastomeres lose the ability to induce mesoderm (Zhang, J., Houston, D. W., King, M. L., Payne, C., Wylie, C. and Heasman, J. (1998) Cell 94, 515-524). The targets of VegT, a transcription activator, must therefore include genes involved both in the specification of endoderm and in the production of mesoderm-inducing signals. We recently reported that the upstream regulatory region of the homeobox-containing gene Bix4 contains T-box binding sites. Here we show that expression of Bix4 requires maternal VegT and that two T box binding sites are necessary and sufficient for mesodermal and endodermal expression of reporter genes driven by the Bix4 promoter in transgenic Xenopus embryos. Remarkably, a single T-box binding site is able to act as a mesoderm specific enhancer when placed upstream of a minimal promoter. Finally, we show that Bix4 rescues the formation of endodermal markers in embryos in which VegT transcripts have been ablated but does not restore the ability of vegetal pole blastomeres to induce mesoderm. These results demonstrate that Bix4 acts directly downstream of VegT to specify endodermal differentiation in Xenopus embryos. PMID- 10477289 TI - Engrailed-1 and netrin-1 regulate axon pathfinding by association interneurons that project to motor neurons. AB - During early development, multiple classes of interneurons are generated in the spinal cord including association interneurons that synapse with motor neurons and regulate their activity. Very little is known about the molecular mechanisms that generate these interneuron cell types, nor is it known how axons from association interneurons are guided toward somatic motor neurons. By targeting the axonal reporter gene &tgr;-lacZ to the En1 locus, we show the cell-type specific transcription factor Engrailed-1 (EN1) defines a population of association neurons that project locally to somatic motor neurons. These EN1 interneurons are born early and their axons pioneer an ipsilateral longitudinal projection in the ventral spinal cord. The EN1 interneurons extend axons in a stereotypic manner, first ventrally, then rostrally for one to two segments where their axons terminate close to motor neurons. We show that the growth of EN1 axons along a ventrolateral pathway toward motor neurons is dependent on netrin-1 signaling. In addition, we demonstrate that En1 regulates pathfinding and fasciculation during the second phase of EN1 axon growth in the ventrolateral funiculus (VLF); however, En1 is not required for the early specification of ventral interneuron cell types in the embryonic spinal cord. PMID- 10477290 TI - Pax6 induces ectopic eyes in a vertebrate. AB - We report here that misexpression of the transcription factor Pax6 in the vertebrate Xenopus laevis leads to the formation of differentiated ectopic eyes. Multiple molecular markers indicated the presence of mature lens fiber cells, ganglion cells, Muller cells, photoreceptors and retinal pigment epithelial cells in a spatial arrangement similar to that of endogenous eyes. Lineage tracing experiments showed that lens, retina and retinal pigment epithelium arose as a consequence of the cell-autonomous function of Pax6. These experiments also reveal that the cell autonomous activity of misexpressed Pax6 causes the ectopic expression of a number of genes including Rx, Otx2, Six3 and endogenous Pax6, each of which has been implicated in eye development. The formation of ectopic and endogenous eyes could be suppressed by coexpression of a dominant-negative form of Pax6. These data show that in vertebrates, as in the invertebrate Drosophila melanogaster, Pax6 is both necessary and sufficient to trigger the cascade of events required for eye formation. PMID- 10477291 TI - Control of segmental expression of the cardiac-restricted ankyrin repeat protein gene by distinct regulatory pathways in murine cardiogenesis. AB - Although accumulating evidence suggests that the heart develops in a segmental fashion, the molecular mechanisms that control regional specification of cardiomyocytes in the developing heart remain largely unknown. In this study, we have used the mouse cardiac-restricted ankyrin repeat protein (CARP) gene as a model system to study these mechanisms. The CARP gene encodes a nuclear co regulator for cardiac gene expression, which lies downstream of the cardiac homeobox gene, Nkx 2.5, and is an early marker of the cardiac muscle cell lineage. We have demonstrated that the expression of the gene is developmentally down regulated and dramatically induced as part of the embryonic gene program during cardiac hypertrophy. Using a lacZ/knock-in mouse and three lines of transgenic mouse harboring various CARP promoter/lacZ reporters, we have identified distinct 5' cis regulatory elements of the gene that can direct heart segment-specific transgene expression, such as atrial versus ventricular and left versus right. Most interestingly, a 213 base pair sequence element of the gene was found to confer conotruncal segment-specific transgene expression. Using the transgene as a conotruncal segment-specific marker, we were able to document the developmental fate of a subset of cardiomyocytes in the conotruncus during cardiogenesis. In addition, we have identified an essential GATA-4 binding site in the proximal upstream regulatory region of the gene and cooperative transcriptional regulation mediated by Nkx2.5 and GATA-4. We have shown that this cooperative regulation is dependent on binding of GATA-4 to its cognate DNA sequence in the promoter, which suggests that Nkx2.5 controls CARP expression, at least in part, through GATA-4. PMID- 10477292 TI - Shade avoidance responses are mediated by the ATHB-2 HD-zip protein, a negative regulator of gene expression. AB - The ATHB-2 gene encoding an homeodomain-leucine zipper protein is rapidly and strongly induced by changes in the ratio of red to far-red light which naturally occur during the daytime under the canopy and induce in many plants the shade avoidance response. Here, we show that elevated ATHB-2 levels inhibit cotyledon expansion by restricting cell elongation in the cotyledon-length and -width direction. We also show that elevated ATHB-2 levels enhance longitudinal cell expansion in the hypocotyl. Interestingly, we found that ATHB-2-induced, as well as shade-induced, elongation of the hypocotyl is dependent on the auxin transport system. In the root and hypocotyl, elevated ATHB-2 levels also inhibit specific cell proliferation such as secondary growth of the vascular system and lateral root formation. Consistent with the key role of auxin in these processes, we found that auxin is able to rescue the ATHB-2 lateral root phenotype. We also show that reduced levels of ATHB-2 result in reciprocal phenotypes. Moreover, we demonstrate that ATHB-2 functions as a negative regulator of gene expression in a transient assay. Remarkably, the expression in transgenic plants of a derivative of ATHB-2 with the same DNA binding specificity but opposite regulatory properties results in a shift in the orientation of hypocotyl cell expansion toward radial expansion, and in an increase in hypocotyl secondary cell proliferation. A model of ATHB-2 function in the regulation of shade-induced growth responses is proposed. PMID- 10477293 TI - Transplacental delivery of the Wnt antagonist Frzb1 inhibits development of caudal paraxial mesoderm and skeletal myogenesis in mouse embryos. AB - Axial structures (neural tube/notochord) and surface ectoderm activate myogenesis in the mouse embryo; their action can be reproduced, at least in part, by several molecules such as Sonic hedgehog and Wnts. Recently, soluble Wnt antagonists have been identified. Among those examined only Frzb1 was found to be expressed in the presomitic mesoderm and newly formed somites and thus its possible role in regulating myogenesis was investigated in detail. When presomitic mesoderm or newly formed somites were cultured with axial structures and surface ectoderm on a feeder layer of C3H10T1/2 cells expressing Frzb1, myogenesis was abolished or severely reduced in presomitic mesoderm and the three most recently formed somites. In contrast, no effect was observed on more mature somites. Inhibition of myogenesis did not appear to be associated with increased cell death since the final number of cells in the explants grown in the presence of Frzb1 was only slightly reduced in comparison with controls. In order to examine the possible function of Frzb1 in vivo, we developed a method based on the overexpression of the soluble antagonist by transient transfection of WOP cells with a Frzb1 expression vector and injection of transfected cells into the placenta of pregnant females before the onset of maternofoetal circulation. Frzb1, secreted by WOP cells, accumulated in the embryo and caused a marked reduction in size of caudal structures. Myogenesis was strongly reduced and, in the most severe cases, abolished. This was not due to a generalized toxic effect since only several genes downstream of the Wnt signaling pathway such as En1, Noggin and Myf5 were downregulated; in contrast, Pax3 and Mox1 expression levels were not affected even in embryos exhibiting the most severe phenotypes. Taken together, these results suggest that Wnt signals may act by regulating both myogenic commitment and expansion of committed cells in the mouse mesoderm. PMID- 10477294 TI - Functional association of retinoic acid and hedgehog signaling in Xenopus primary neurogenesis. AB - Previous work has shown that the posteriorising agent retinoic acid can accelerate anterior neuronal differentiation in Xenopus laevis embryos (Papalopulu, N. and Kintner, C. (1996) Development 122, 3409-3418). To elucidate the role of retinoic acid in the primary neurogenesis cascade, we investigated whether retinoic acid treatment of whole embryos could change the spatial expression of a set of genes known to be involved in neurogenesis. We show that retinoic acid expands the N-tubulin, X-ngnr-1, X-MyT1, X-&Dgr;-1 and Gli3 domains and inhibits the expression of Zic2 and sonic hedgehog in the neural ectoderm, whereas a retinoid antagonist produces opposite changes. In contrast, sonic and banded hedgehog overexpression reduced the N-tubulin stripes, enlarged the neural plate at the expense of the neural crest, downregulated Gli3 and upregulated Zic2. Thus, retinoic acid and hedgehog signaling have opposite effects on the prepattern genes Gli3 and Zic2 and on other genes acting downstream in the neurogenesis cascade. In addition, retinoic acid cannot rescue the inhibitory effect of Notch(ICD), Zic2 or sonic hedgehog on primary neurogenesis. Our results suggest that retinoic acid acts very early, upstream of sonic hedgehog, and we propose a model for regulation of differentiation and proliferation in the neural plate, showing that retinoic acid might be activating primary neurogenesis by repressing sonic hedgehog expression. PMID- 10477295 TI - Inhibition of TGF-beta receptor signaling in osteoblasts leads to decreased bone remodeling and increased trabecular bone mass. AB - Transforming growth factor-beta (TGF-beta) is abundant in bone matrix and has been shown to regulate the activity of osteoblasts and osteoclasts in vitro. To explore the role of endogenous TGF-(beta) in osteoblast function in vivo, we have inhibited osteoblastic responsiveness to TGF-beta in transgenic mice by expressing a cytoplasmically truncated type II TGF-beta receptor from the osteocalcin promoter. These transgenic mice develop an age-dependent increase in trabecular bone mass, which progresses up to the age of 6 months, due to an imbalance between bone formation and resorption during bone remodeling. Since the rate of osteoblastic bone formation was not altered, their increased trabecular bone mass is likely due to decreased bone resorption by osteoclasts. Accordingly, direct evidence of reduced osteoclast activity was found in transgenic mouse skulls, which had less cavitation and fewer mature osteoclasts relative to skulls of wild-type mice. These bone remodeling defects resulted in altered biomechanical properties. The femurs of transgenic mice were tougher, and their vertebral bodies were stiffer and stronger than those of wild-type mice. Lastly, osteocyte density was decreased in transgenic mice, suggesting that TGF-beta signaling in osteoblasts is required for normal osteoblast differentiation in vivo. Our results demonstrate that endogenous TGF-beta acts directly on osteoblasts to regulate bone remodeling, structure and biomechanical properties. PMID- 10477296 TI - The Drosophila chiffon gene is required for chorion gene amplification, and is related to the yeast Dbf4 regulator of DNA replication and cell cycle. AB - The Drosophila chorion genes encode the major protein components of the chorion (eggshell) and are arranged in two clusters in the genome. To meet the demand for rapid chorion synthesis, Drosophila ovary follicle cells amplify the chorion gene clusters approximately 80-fold. Amplification proceeds through repeated firing of one or more DNA replication origins located near the center of each gene cluster. Hypomorphic mutant alleles of the chiffon gene cause thin, fragile chorions and female sterility, and were found to eliminate chorion gene amplification. Null alleles of chiffon had the additional phenotypes of rough eyes and thin thoracic bristles: phenotypes often associated with disruption of normal cell cycle. The chiffon locus was cloned by chromosomal walking from the nearby cactus locus. A 6.5 kb transcript was identified and confirmed to be chiffon by sequencing of mutant alleles and by phenotypic rescue with genomic transformation constructs. The protein predicted by translation of the 5.1 kb chiffon ORF contains two domains related to the S. cerevisiae Dbf4 regulator of DNA replication origin firing and cell cycle progression: a 44 residue domain designated CDDN1 (43% identical) and a 41 residue domain designated CDDN2 (12% identical). The CDDN domains were also found in the S. pombe homolog of Dbf4, Dfp1, as well as in the proteins predicted by translation of the Aspergillus nimO gene and specific human and mouse clones. The data suggest a family of eukaryotic proteins related to Dbf4 and involved in initiation of DNA replication. PMID- 10477297 TI - Cell autonomous and non-cell autonomous functions of Otx2 in patterning the rostral brain. AB - Previous studies have shown that the homeobox gene Otx2 is required first in the visceral endoderm for induction of forebrain and midbrain, and subsequently in the neurectoderm for its regional specification. Here, we demonstrate that Otx2 functions both cell autonomously and non-cell autonomously in neurectoderm cells of the forebrain and midbrain to regulate expression of region-specific homeobox and cell adhesion genes. Using chimeras containing both Otx2 mutant and wild-type cells in the brain, we observe a reduction or loss of expression of Rpx/Hesx1, Wnt1, R-cadherin and ephrin-A2 in mutant cells, whereas expression of En2 and Six3 is rescued by surrounding wild-type cells. Forebrain Otx2 mutant cells subsequently undergo apoptosis. Altogether, this study demonstrates that Otx2 is an important regulator of brain patterning and morphogenesis, through its regulation of candidate target genes such as Rpx/Hesx1, Wnt1, R-cadherin and ephrin-A2. PMID- 10477298 TI - Characterization of the early development of specific hypaxial muscles from the ventrolateral myotome. AB - We have previously found that the myotome is formed by a first wave of pioneer cells generated along the medial epithelial somite and a second wave emanating from the dorsomedial lip (DML), rostral and caudal edges of the dermomyotome (Kahane, N., Cinnamon, Y. and Kalcheim, C. (1998a) Mech. Dev. 74, 59-73; Kahane, N., Cinnamon, Y. and Kalcheim, C. (1998b) Development 125, 4259-4271). In this study, we have addressed the development and precise fate of the ventrolateral lip (VLL) in non-limb regions of the axis. To this end, fluorescent vital dyes were iontophoretically injected in the center of the VLL and the translocation of labeled cells was followed by confocal microscopy. VLL-derived cells colonized the ventrolateral portion of the myotome. This occurred following an early longitudinal cell translocation along the medial boundary until reaching the rostral or caudal dermomyotome lips from which fibers emerged into the myotome. Thus, the behavior of VLL cells parallels that of their DML counterparts which colonize the opposite, dorsomedial portion of the myotome. To precisely understand the way the myotome expands, we addressed the early generation of hypaxial intercostal muscles. We found that intercostal muscles were formed by VLL-derived fibers that intermingled with fibers emerging from the ventrolateral aspect of both rostral and caudal edges of the dermomyotome. Notably, hypaxial intercostal muscles also contained pioneer myofibers (first wave) showing for the first time that lateral myotome-derived muscles contain a fundamental component of fibers generated in the medial domain of the somite. In addition, we show that during myotome growth and evolution into muscle, second-wave myofibers progressively intercalate between the pioneer fibers, suggesting a constant mode of myotomal expansion in its dorsomedial to ventrolateral extent. This further suggests that specific hypaxial muscles develop following a consistent ventral expansion of a 'compound myotome' into the somatopleure. PMID- 10477299 TI - Dual role of the basic helix-loop-helix transcription factor scleraxis in mesoderm formation and chondrogenesis during mouse embryogenesis. AB - Scleraxis is a basic helix-loop-helix (bHLH) transcription factor shown previously to be expressed in developing chondrogenic cell lineages during embryogenesis. To investigate its function in embryonic development, we produced scleraxis-null mice by gene targeting. Homozygous mutant embryos developed normally until the early egg cylinder stage (embryonic day 6.0), when they became growth-arrested and failed to gastrulate. Consistent with this early embryonic phenotype, scleraxis was found to be expressed throughout the embryo at the time of gastrulation before becoming restricted to chondrogenic precursor cells at embryonic day 9.5. At the time of developmental arrest, scleraxis-null embryos consisted of ectodermal and primitive endodermal cell layers, but lacked a primitive streak or recognizable mesoderm. Analysis of molecular markers of the three embryonic germ layers confirmed that scleraxis mutant embryos were unable to form mesoderm. By generating chimeric embryos, using lacZ-marked scleraxis null and wild-type embryonic stem cells, we examined the ability of mutant cells to contribute to regions of the embryo beyond the time of lethality of homozygous mutants. Scleraxis-null cells were specifically excluded from the sclerotomal compartment of somites, which gives rise to the axial skeleton, and from developing ribs, but were able to contribute to most other regions of the embryo, including mesoderm-derived tissues. These results reveal an essential early role for scleraxis in mesoderm formation, as well as a later role in formation of somite-derived chondrogenic lineages, and suggest that scleraxis target genes mediate these processes. PMID- 10477300 TI - Proteolysis of cubitus interruptus in Drosophila requires phosphorylation by protein kinase A. AB - The Hedgehog signal transduction pathway is involved in diverse patterning events in many organisms. In Drosophila, Hedgehog signaling regulates transcription of target genes by modifying the activity of the DNA-binding protein Cubitus interruptus (Ci). Hedgehog signaling inhibits proteolytic cleavage of full-length Ci (Ci-155) to Ci-75, a form that represses some target genes, and also converts the full-length form to a potent transcriptional activator. Reduction of protein kinase A (PKA) activity also leads to accumulation of full-length Ci and to ectopic expression of Hedgehog target genes, prompting the hypothesis that PKA might normally promote cleavage to Ci-75 by directly phosphorylating Ci-155. Here we show that a mutant form of Ci lacking five potential PKA phosphorylation sites (Ci5m) is not detectably cleaved to Ci-75 in Drosophila embryos. Moreover, changes in PKA activity dramatically altered levels of full-length wild-type Ci in embryos and imaginal discs, but did not significantly alter full-length Ci5m levels. We corroborate these results by showing that Ci5m is more active than wild-type Ci at inducing ectopic transcription of the Hh target gene wingless in embryos and that inhibition of PKA enhances induction of wingless by wild-type Ci but not by Ci5m. We therefore propose that PKA phosphorylation of Ci is required for the proteolysis of Ci-155 to Ci-75 in vivo. We also show that the activity of Ci5m remains Hedgehog responsive if expressed at low levels, providing further evidence that the full-length form of Ci undergoes a Hedgehog-dependent activation step. PMID- 10477301 TI - The Drosophila dead ringer gene is required for early embryonic patterning through regulation of argos and buttonhead expression. AB - The dead ringer (dri) gene of Drosophila melanogaster is a member of the recently discovered ARID-box family of eukaryotic genes that encode proteins with a conserved DNA binding domain. dri itself is highly conserved, with specific orthologs in the human, mouse, zebrafish and C. elegans genomes. We have generated dri mutant alleles to show that dri is essential for anterior-posterior patterning and for muscle development in the embryo. Consistent with the mutant phenotype and the sequence-specific DNA-binding properties of its product, dri was found to be essential for the normal early embryonic expression pattern of several key regulatory genes. In dri mutant embryos, expression of argos in the terminal domains was severely reduced, accounting for the dri mutant head phenotype. Conversely, buttonhead expression was found to be deregulated in the trunk region, accounting for the appearance of ectopic cephalic furrows. Curiously, dri was found also to be required for maintenance of expression of the ventrolateral region of even-skipped stripe four. This study establishes dri as an essential co-factor in the regulated expression of specific patterning genes during early embryogenesis. PMID- 10477302 TI - In vivo transplantation of mammalian neural crest cells into chick hosts reveals a new autonomic sublineage restriction. AB - The study of mammalian neural crest development has been limited by the lack of an accessible system for in vivo transplantation of these cells. We have developed a novel transplantation system to study lineage restriction in the rodent neural crest. Migratory rat neural crest cells (NCCs), transplanted into chicken embryos, can differentiate into sensory, sympathetic, and parasympathetic neurons, as shown by the expression of neuronal subtype-specific and pan-neuronal markers, as well as into Schwann cells and satellite glia. In contrast, an immunopurified population of enteric neural precursors (ENPs) from the fetal gut can also generate neurons in all of these ganglia, but only expresses appropriate neuronal subtype markers in Remak's and associated pelvic parasympathetic ganglia. ENPs also appear restricted in the kinds of glia they can generate in comparison to NCCs. Thus ENPs have parasympathetic and presumably enteric capacities, but not sympathetic or sensory capacities. These results identify a new autonomic lineage restriction in the neural crest, and suggest that this restriction preceeds the choice between neuronal and glial fates. PMID- 10477303 TI - Developmental changes in the response of trigeminal neurons to neurotrophins: influence of birthdate and the ganglion environment. AB - Previous studies have shown that most neurons in cultures established during the early stages of neurogenesis in the embryonic mouse trigeminal ganglion are supported by BDNF whereas most neurons cultured from older ganglia survive with NGF. To ascertain to what extent these developmental changes in neurotrophin responsiveness result from separate phases of generation of BDNF- and NGF responsive neurons or from a developmental switch in the response of neurons from BDNF to NGF, we administered BrdU to pregnant mice at different stages of gestation to identify neurons born at different times and studied the survival of labelled neurons in dissociated cultures established shortly after BrdU administration. Most early-generated neurons responded to BDNF, neurons generated at intermediate times responded to both factors and late-generated neurons responded to NGF, indicating that there are overlapping phases in the generation of BDNF- and NGF-responsive neurons and that late-generated neurons do not switch responsiveness from BDNF to NGF. To ascertain if early-generated neurons do switch their response to neurotrophins during development, we used repeated BrdU injection to label all neurons generated after an early stage in neurogenesis and studied the neurotrophin responsiveness of the unlabelled neurons in cultures established after neurogenesis had ceased. The response of these early-generated neurons had decreased to BDNF and increased to NGF, indicating that at least a proportion of early-generated neurons switch responsiveness to neurotrophins in vivo. Because early-generated neurons do not switch responsiveness from BDNF to NGF in long-term dissociated cultures, we cultured early trigeminal ganglion explants with and without their targets for 24 hours before establishing dissociated cultures. This period of explant culture was sufficient to enable many early-generated neurons to switch their response from BDNF to NGF and this switch occurred irrespective of presence of target tissue. Our findings conclusively demonstrate for the first time that individual neurons switch their neurotrophin requirements during development and that this switch depends on cell interactions within the ganglion. In addition, we show that there are overlapping phases in the generation of BDNF- and NGF-responsive neurons in the trigeminal ganglion. PMID- 10477304 TI - Directionality of wingless protein transport influences epidermal patterning in the Drosophila embryo. AB - Active endocytotic processes are required for the normal distribution of Wingless (Wg) protein across the epidermal cells of each embryonic segment. To assess the functional consequences of this broad Wg distribution, we have devised a means of perturbing endocytosis in spatially restricted domains within the embryo. We have constructed a transgene expressing a dominant negative form of shibire (shi), the fly dynamin homologue. When this transgene is expressed using the GAL4-UAS system, we find that Wg protein distribution within the domain of transgene expression is limited and that Wg-dependent epidermal patterning events surrounding the domain of expression are disrupted in a directional fashion. Our results indicate that Wg transport in an anterior direction generates the normal expanse of naked cuticle within the segment and that movement of Wg in a posterior direction specifies diverse denticle cell fates in the anterior portion of the adjacent segment. Furthermore, we have discovered that interfering with posterior movement of Wg rescues the excessive naked cuticle specification observed in naked (nkd) mutant embryos. We propose that the nkd segment polarity phenotype results from unregulated posterior transport of Wg protein and therefore that wild-type Nkd function may contribute to the control of Wg movement within the epidermal cells of the segment. PMID- 10477305 TI - Head versus trunk patterning in the Drosophila embryo; collier requirement for formation of the intercalary segment. AB - Whereas the segmental nature of the insect head is well established, relatively little is known about the genetic and molecular mechanisms governing this process. In this paper, we report the phenotypic analysis of mutations in collier (col), which encodes the Drosophila member of the COE family of HLH transcription factors and is activated at the blastoderm stage in a region overlapping a parasegment (PS0: posterior intercalary and anterior mandibular segments) and a mitotic domain, MD2. col mutant embryos specifically lack intercalary ectodermal structures. col activity is required for intercalary-segment expression both of the segment polarity genes hedgehog, engrailed, and wingless, and of the segment identity gene cap and collar. The parasegmental register of col activation is controlled by the combined activities of the head-gap genes buttonhead and empty spiracles and the pair-rule gene even skipped; it therefore integrates inputs from both the head and trunk segmentation systems, which were previously considered as being essentially independent. After gastrulation, positive autoregulation of col is limited to cells of anterior PS0. Conversely, heat-pulse induced ubiquitous expression of Col leads to disruption of the head skeleton. Together, these results indicate that col is required for establishment of the PS(-1)/PS0 parasegmental border and formation of the intercalary segment. Our data support neither a simple combinatorial model for segmental patterning of the head nor a direct activation of segment polarity gene expression by head-gap genes, but rather argue for the existence of parasegment-specific second order regulators acting in the head, at a level similar to that of pair-rule genes in the trunk. PMID- 10477306 TI - Transposon Tn21, flagship of the floating genome. AB - The transposon Tn21 and a group of closely related transposons (the Tn21 family) are involved in the global dissemination of antibiotic resistance determinants in gram-negative facultative bacteria. The molecular basis for their involvement is carriage by the Tn21 family of a mobile DNA element (the integron) encoding a site-specific system for the acquisition of multiple antibiotic resistance genes. The paradigm example, Tn21, also carries genes for its own transposition and a mercury resistance (mer) operon. We have compiled the entire 19,671-bp sequence of Tn21 and assessed the possible origins and functions of the genes it contains. Our assessment adds molecular detail to previous models of the evolution of Tn21 and is consistent with the insertion of the integron In2 into an ancestral Tn501 like mer transposon. Codon usage analysis indicates distinct host origins for the ancestral mer operon, the integron, and the gene cassette and two insertion sequences which lie within the integron. The sole gene of unknown function in the integron, orf5, resembles a puromycin-modifying enzyme from an antibiotic producing bacterium. A possible seventh gene in the mer operon (merE), perhaps with a role in Hg(II) transport, lies in the junction between the integron and the mer operon. Analysis of the region interrupted by insertion of the integron suggests that the putative transposition regulator, tnpM, is the C-terminal vestige of a tyrosine kinase sensor present in the ancestral mer transposon. The extensive dissemination of the Tn21 family may have resulted from the fortuitous association of a genetic element for accumulating multiple antibiotic resistances (the integron) with one conferring resistance to a toxic metal at a time when clinical, agricultural, and industrial practices were rapidly increasing the exposure to both types of selective agents. The compendium offered here will provide a reference point for ongoing observations of related elements in multiply resistant strains emerging worldwide. PMID- 10477307 TI - Genetics of O-antigen biosynthesis in Pseudomonas aeruginosa. AB - Pathogenic bacteria produce an elaborate assortment of extracellular and cell associated bacterial products that enable colonization and establishment of infection within a host. Lipopolysaccharide (LPS) molecules are cell surface factors that are typically known for their protective role against serum-mediated lysis and their endotoxic properties. The most heterogeneous portion of LPS is the O antigen or O polysaccharide, and it is this region which confers serum resistance to the organism. Pseudomonas aeruginosa is capable of concomitantly synthesizing two types of LPS referred to as A band and B band. The A-band LPS contains a conserved O polysaccharide region composed of D-rhamnose (homopolymer), while the B-band O-antigen (heteropolymer) structure varies among the 20 O serotypes of P. aeruginosa. The genes coding for the enzymes that direct the synthesis of these two O antigens are organized into two separate clusters situated at different chromosomal locations. In this review, we summarize the organization of these two gene clusters to discuss how A-band and B-band O antigens are synthesized and assembled by dedicated enzymes. Examples of unique proteins required for both A-band and B-band O-antigen synthesis and for the synthesis of both LPS and alginate are discussed. The recent identification of additional genes within the P. aeruginosa genome that are homologous to those in the A-band and B-band gene clusters are intriguing since some are able to influence O-antigen synthesis. These studies demonstrate that P. aeruginosa represents a unique model system, allowing studies of heteropolymeric and homopolymeric O-antigen synthesis, as well as permitting an examination of the interrelationship of the synthesis of LPS molecules and other virulence determinants. PMID- 10477308 TI - Function and regulation of yeast hexose transporters. AB - Glucose, the most abundant monosaccharide in nature, is the principal carbon and energy source for nearly all cells. The first, and rate-limiting, step of glucose metabolism is its transport across the plasma membrane. In cells of many organisms glucose ensures its own efficient metabolism by serving as an environmental stimulus that regulates the quantity, types, and activity of glucose transporters, both at the transcriptional and posttranslational levels. This is most apparent in the baker's yeast Saccharomyces cerevisiae, which has 20 genes encoding known or likely glucose transporters, each of which is known or likely to have a different affinity for glucose. The expression and function of most of these HXT genes is regulated by different levels of glucose. This review focuses on the mechanisms S. cerevisiae and a few other fungal species utilize for sensing the level of glucose and transmitting this information to the nucleus to alter HXT gene expression. One mechanism represses transcription of some HXT genes when glucose levels are high and works through the Mig1 transcriptional repressor, whose function is regulated by the Snf1-Snf4 protein kinase and Reg1 Glc7 protein phosphatase. Another pathway induces HXT expression in response to glucose and employs the Rgt1 transcriptional repressor, a ubiquitin ligase protein complex (SCF(Grr1)) that regulates Rgt1 function, and two glucose sensors in the membrane (Snf3 and Rgt2) that bind glucose and generate the intracellular signal to which Rgt1 responds. These two regulatory pathways collaborate with other, less well-understood, pathways to ensure that yeast cells express the glucose transporters best suited for the amount of glucose available. PMID- 10477310 TI - Gliding motility in bacteria: insights from studies of Myxococcus xanthus. AB - Gliding motility is observed in a large variety of phylogenetically unrelated bacteria. Gliding provides a means for microbes to travel in environments with a low water content, such as might be found in biofilms, microbial mats, and soil. Gliding is defined as the movement of a cell on a surface in the direction of the long axis of the cell. Because this definition is operational and not mechanistic, the underlying molecular motor(s) may be quite different in diverse microbes. In fact, studies on the gliding bacterium Myxococcus xanthus suggest that two independent gliding machineries, encoded by two multigene systems, operate in this microorganism. One machinery, which allows individual cells to glide on a surface, independent of whether the cells are moving alone or in groups, requires the function of the genes of the A-motility system. More than 37 A-motility genes are known to be required for this form of movement. Depending on an additional phenotype, these genes are divided into two subclasses, the agl and cgl genes. Videomicroscopic studies on gliding movement, as well as ultrastructural observations of two myxobacteria, suggest that the A-system motor may consist of multiple single motor elements that are arrayed along the entire cell body. Each motor element is proposed to be localized to the periplasmic space and to be anchored to the peptidoglycan layer. The force to glide which may be generated here is coupled to adhesion sites that move freely in the outer membrane. These adhesion sites provide a specific contact with the substratum. Based on single-cell observations, similar models have been proposed to operate in the unrelated gliding bacteria Flavobacterium johnsoniae (formerly Cytophaga johnsonae), Cytophaga strain U67, and Flexibacter polymorphus (a filamentous glider). Although this model has not been verified experimentally, M. xanthus seems to be the ideal organism with which to test it, given the genetic tools available. The second gliding motor in M. xanthus controls cell movement in groups (S-motility system). It is dependent on functional type IV pili and is operative only when cells are in close proximity to each other. Type IV pili are known to be involved in another mode of bacterial surface translocation, called twitching motility. S-motility may well represent a variation of twitching motility in M. xanthus. However, twitching differs from gliding since it involves cell movements that are jerky and abrupt and that lack the organization and smoothness observed in gliding. Components of this motor are encoded by genes of the S-system, which appear to be homologs of genes involved in the biosynthesis, assembly, and function of type IV pili in Pseudomonas aeruginosa and Neisseria gonorrhoeae. How type IV pili generate force in S-motility is currently unknown, but it is to be expected that ongoing physiological, genetic, and biochemical studies in M. xanthus, in conjunction with studies on twitching in P. aeruginosa and N. gonorrhoeae, will provide important insights into this microbial motor. The two motility systems of M. xanthus are affected to different degrees by the MglA protein, which shows similarity to a small GTPase. Bacterial chemotaxis-like sensory transduction systems control gliding motility in M. xanthus. The frz genes appear to regulate gliding movement of individual cells and movement by the S-motility system, suggesting that the two motors found in this bacterium can be regulated to result in coordinated multicellular movements. In contrast, the dif genes affect only S-system-dependent swarming. PMID- 10477313 TI - Fungal resistance to plant antibiotics as a mechanism of pathogenesis. AB - Many plants produce low-molecular-weight compounds which inhibit the growth of phytopathogenic fungi in vitro. These compounds may be preformed inhibitors that are present constitutively in healthy plants (also known as phytoanticipins), or they may be synthesized in response to pathogen attack (phytoalexins). Successful pathogens must be able to circumvent or overcome these antifungal defenses, and this review focuses on the significance of fungal resistance to plant antibiotics as a mechanism of pathogenesis. There is increasing evidence that resistance of fungal pathogens to plant antibiotics can be important for pathogenicity, at least for some fungus-plant interactions. This evidence has emerged largely from studies of fungal degradative enzymes and also from experiments in which plants with altered levels of antifungal secondary metabolites were generated. Whereas the emphasis to date has been on degradative mechanisms of resistance of phytopathogenic fungi to antifungal secondary metabolites, in the future we are likely to see a rapid expansion in our knowledge of alternative mechanisms of resistance. These may include membrane efflux systems of the kind associated with multidrug resistance and innate resistance due to insensitivity of the target site. The manipulation of plant biosynthetic pathways to give altered antibiotic profiles will also be valuable in telling us more about the significance of antifungal secondary metabolites for plant defense and clearly has great potential for enhancing disease resistance for commercial purposes. PMID- 10477322 TI - Evaluation of aminoalkylmethacrylate nanoparticles as colloidal drug carrier systems. Part II: characterization of antisense oligonucleotides loaded copolymer nanoparticles. AB - Aminoalkylmethacrylate methylmethacrylate copolymer nanoparticles were evaluated for their use as potential drug carrier systems. Their cytotoxicity, as well as the loading of antisense oligonucleotides that were employed as anionic model drugs depended on the substitution of the basic aminoalkyl copolymer. Toxic influences on the integrity of cell membranes depended on aminoalkyl groups located on the particle surfaces. Toxicity was observed either by 3-[4,5 dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assays using African green monkey kidney (AGMK) cells or by a hemolysis test, where the efflux of haemoglobin from disrupted erythrocytes was measured. The cytotoxic effects were increased by the elongation of the N-alkyl chain by four additional methylene groups. Lipophilic polymethylmethacrylate (PMMA) homopolymer nanoparticles showed a negative surface charge and, therefore, were not suitable for the adsorption of anionic drugs. The surface charge was changed to positive values by the incorporation of basic monomers. Consequently, the loading efficacy was increased by raising the basic copolymer portion. Additionally, a pH-dependent loading behaviour of oligonucleotides was observed. Substitution of the amino nitrogen protons by methyl groups led to a decreased oligonucleotide loading and to a reduced cytotoxicity. Nanoparticles with permanent positively charged quarternary ammonium groups showed a high pH-independent loading efficacy, but also possessed a high cytotoxic potential. In this study, cationic copolymer nanoparticles containing 30% (w/w) methylaminoethyl-methacrylate (MMAEMC) were found to be optimal with regard to biocompatibility and carrier properties for hydrophilic anionic antisense oligonucleotides. A significant portion of adsorbed oligonucleotides were protected from enzymatic degradation. The cellular uptake of oligonucleotides into Vero cells was significantly enhanced by this methylaminoethyl-methacrylate derivative. PMID- 10477323 TI - In vitro/in vivo correlations of dissolution data of carbamazepine immediate release tablets with pharmacokinetic data obtained in healthy volunteers. AB - The aim of the study was to select a dissolution test method for carbamazepine (CBZ) immediate release tablets, giving the best in vitro/in vivo correlations (IVIVC) and to determine the potential of this method as an estimate for bioequivalence testing. Four 200 mg CBZ products which are sold on the Dutch market, covering the innovator and three generic products, were selected. They had been tested in a randomised, fourway cross-over bioavailability study in healthy volunteers. Their dissolution rate behaviour in vitro was investigated in two dissolution media: (1) 1% sodium lauryl sulphate in water (SLS), in accordance with the United States Pharmacopeia (USP); (2) 0.1 mol/l Hydrochloric acid in water (HC). In the bioavailability study these products had shown no large differences in the extent of absorption (AUC(0-infinity);) but large differences in absorption rate. The products now also showed large differences in dissolution rate in vitro in both dissolution media, the rank order being the same as for the absorption rate. It was concluded that the absorption rate in vivo depends on the dissolution rate in vivo. 'Level C' IVIVC according to the USP were optimised by plotting percentages dissolved on selected time points (D values) or their reciprocals (1/D values), against several pharmacokinetic parameters primarily related to the absorption phase and against AUC(0-infinity). In this way for each IVIVC the optimum D or 1/D value, was calculated. For both media no meaningful IVIVC were obtained with AUC(0-infinity), but favourable IVIVC were obtained with the parameters primarily related to the absorption phase. In the bioavailability study indicated above it was found that, among the pharmacokinetic characteristics primarily related to the absorption phase, C(max) is the most promising in expressing rate of absorption in bioequivalence testing in single dose studies with CBZ immediate release tablets. Consequently, C(max) was selected for expressing rate of absorption. The most favourable IVIVC were obtained with D(20) in SLS versus C(max). From this IVIVC and the requirements for bioequivalence (AUC(0-infinity): 0.8-1.25 and C(max) : 0.75-1.35; 90% confidence interval), a specification for dissolution testing in SLS was calculated as follows: 'after 20 minutes, 34-99% dissolved'. Owing to the fact that the rate of absorption in vivo depends on i.a. the dissolution rate in vivo, it can be concluded that with this specification bioequivalence with respect to both rate of absorption and extent of absorption is ensured. As this specification is comparable with the USP specification: 'not less than 75% dissolved after 1 h', it is concluded that the USP specification is suitable to ensure bioequivalence of CBZ immediate release tablets. PMID- 10477312 TI - Helicobacter pylori physiology predicted from genomic comparison of two strains. AB - Helicobacter pylori is a gram-negative bacteria which colonizes the gastric mucosa of humans and is implicated in a wide range of gastroduodenal diseases. This paper reviews the physiology of this bacterium as predicted from the sequenced genomes of two unrelated strains and reconciles these predictions with the literature. In general, the predicted capabilities are in good agreement with reported experimental observations. H. pylori is limited in carbohydrate utilization and will use amino acids, for which it has transporter systems, as sources of carbon. Energy can be generated by fermentation, and the bacterium possesses components necessary for both aerobic and anaerobic respiration. Sulfur metabolism is limited, whereas nitrogen metabolism is extensive. There is active uptake of DNA via transformation and ample restriction-modification activities. The cell contains numerous outer membrane proteins, some of which are porins or involved in iron uptake. Some of these outer membrane proteins and the lipopolysaccharide may be regulated by a slipped-strand repair mechanism which probably results in phase variation and plays a role in colonization. In contrast to a commonly held belief that H. pylori is a very diverse species, few differences were predicted in the physiology of these two unrelated strains, indicating that host and environmental factors probably play a significant role in the outcome of H. pylori-related disease. PMID- 10477314 TI - Role of alpha(v) integrins in adenovirus cell entry and gene delivery. AB - Adenoviruses (Ad) are a significant cause of acute infections in humans; however, replication-defective forms of this virus are currently under investigation for human gene therapy. Approximately 20 to 25% of all the gene therapy trials (phases I to III) conducted over the past 10 years involve the use of Ad gene delivery for treatment inherited or acquired diseases. At present, the most promising applications involve the use of Ad vectors to irradicate certain nonmetastatic tumors and to promote angiogenesis in order to alleviate cardiovascular disease. While specific problems of using Ad vectors remain to be overcome (as is true for almost all viral and nonviral delivery methods), a distinct advantage of Ad is the extensive knowledge of its macromolecular structure, genome organization, sequence, and mode of replication. Moreover, significant information has also been acquired on the interaction of Ad particles with distinct host cell receptors, events which strongly affect virus tropism. This review provides an overview of the structure and function of Ad attachment (coxsackievirus and Ad receptor [CAR]) and internalization (alpha(v) integrins) receptors and discusses their precise role in virus infection and gene delivery. Recent structure studies of integrin-Ad complexes by cryoelectron microscopy are also highlighted. Finally, unanswered questions arising from the current state of knowledge of Ad-receptor interactions are presented in the context of improving Ad vectors for future human gene therapy applications. PMID- 10477311 TI - Metabolism and genetics of Helicobacter pylori: the genome era. AB - The publication of the complete sequence of Helicobacter pylori 26695 in 1997 and more recently that of strain J99 has provided new insight into the biology of this organism. In this review, we attempt to analyze and interpret the information provided by sequence annotations and to compare these data with those provided by experimental analyses. After a brief description of the general features of the genomes of the two sequenced strains, the principal metabolic pathways are analyzed. In particular, the enzymes encoded by H. pylori involved in fermentative and oxidative metabolism, lipopolysaccharide biosynthesis, nucleotide biosynthesis, aerobic and anaerobic respiration, and iron and nitrogen assimilation are described, and the areas of controversy between the experimental data and those provided by the sequence annotation are discussed. The role of urease, particularly in pH homeostasis, and other specialized mechanisms developed by the bacterium to maintain its internal pH are also considered. The replicational, transcriptional, and translational apparatuses are reviewed, as is the regulatory network. The numerous findings on the metabolism of the bacteria and the paucity of gene expression regulation systems are indicative of the high level of adaptation to the human gastric environment. Arguments in favor of the diversity of H. pylori and molecular data reflecting possible mechanisms involved in this diversity are presented. Finally, we compare the numerous experimental data on the colonization factors and those provided from the genome sequence annotation, in particular for genes involved in motility and adherence of the bacterium to the gastric tissue. PMID- 10477324 TI - Synthesis and characterization of an acylated di-peptide (Myr-Trp-Leu) with modified transmucosal transport properties. AB - In order to improve the buccal absorption of a dipeptide model compound, Tryptophan-Leucine (Trp-Leu), we have synthesised a lipophilic derivative by myristoylation of the N- terminal amino group of Trp-Leu. The acylated peptide (Myr-Trp-Leu) was characterized by HPTLC, purified and isolated by chromatography on a silica gel column. Its structure was confirmed by (13)C and (1)H NMR and mass spectroscopy. The increased lipophilicity of the Myr-Trp-Leu was compared to that of the native peptide by chromatography and by its partition coefficient between n-octanol and saline phosphate buffer. In addition, the sensitivity towards hydrolytic enzymes was studied. The interaction of Trp-Leu with liposomes as model membranes was also studied. The phase transition temperature of dipalmitoylphosphatidylcholine (DPPC) was lowered in the presence of Myr-Trp-Leu, while it was increased in the presence of native parent peptide. Permeation experiments performed in vitro with pig buccal mucosa showed that the Myr-Trp-Leu accumulated in the tissue at the various concentrations tested. In contrast, the native peptide was able to pass through the membrane at all concentrations used. Lipophilic modification of the peptide by acylation drastically changes its behaviour towards tissue systems. PMID- 10477325 TI - The use of carrageenan in mixture with microcrystalline cellulose and its functionality for making tablets. AB - The modulation of tableting and release behavior of combinations of kappa carrageenan Gelcarin((R)) GP 911 NF and microcrystalline cellulose (MCC) Avicel((R)) PH 101 has been evaluated. Graded binary mixtures were tableted to a maximum relative density of 0.850 at the maximum displacement of the upper punch. Additionally, ternary mixtures with the same ratios of kappa-carrageenan and MCC and a constant percentage of theophylline monohydrate (20% (v/v)) were tested for their release behavior. Tablets produced from pure kappa-carrageenan deformed more elastically than pure MCC, the tablets produced were stable but not at the same degree as those made from MCC. Scanning electron microscopy (SEM) pictures showed that for MCC a smooth surface of the tablets resulted, tablets made from kappa-carrageenan showed less 'fusion' and thus more mechanical interlocking is responsible for their stability. Binary mixtures showed a continuous change in compaction properties from plastic to elastic deformation. All ternary mixtures with theophylline deformed more plastically than the binary mixtures, the change in deformation properties remained the same. Theophylline reduced the crushing strength due to its different fracture properties. The ternary mixtures showed different release mechanisms: Fast release up to 20% (v/v) kappa-carrageenan, slower release starting from 30% (v/v). The kinetics of release tended at 70% (v/v) more clearly towards zero-order kinetics. This change in release is in accordance with a change in swelling of tablets made of the binary mixtures. PMID- 10477327 TI - Stability prediction of amorphous benzodiazepines by calculation of the mean relaxation time constant using the Williams-Watts decay function. AB - The enthalpic relaxation of three amorphous benzodiazepines, diazepam, temazepam and triazolam was studied using differential scanning calorimetry for ageing temperatures which were below the glass transition temperature, and ageing times up to 16 h. Experimental determination of the relaxation enthalpy and the heat capacity change, both accompanying the glass transition, enabled us to calculate the extent of relaxation of the amorphous drugs at specific ageing conditions. Fitting of the relaxation function to the Williams-Watts two parameter decay function led to calculation of the mean relaxation time constant tau and the molecular relaxation time distribution parameter beta. The mean relaxation time constants for the three drugs increased from approximately ten h at the glass transition temperature with more than eight orders of magnitude at 66 K below the glass transition temperature. It was found that the benzodiazepines exhibited significant molecular mobility until approximately 50 K below the glass transition temperature; below this temperature molecular mobility becomes unimportant with respect to the shelf life stability. Hence the presented procedure provides the formulation scientist with a tool to set storage conditions for amorphous drugs and glassy pharmaceutical products. PMID- 10477326 TI - Formulation of biphasic release tablets containing slightly soluble drugs. AB - A new biphasic release system for slightly soluble drugs has been proposed. To enhance the dissolution rate, the drug was milled with a superdisintegrant. Then, double-layer tablets were prepared. One layer was formulated to release the drug in a very short time (fast-release). The other consisted of an extended-release hydroxypropylmethylcellulose (HPMC) matrix. Different HPMC concentrations (10, 16 and 22%) and viscosity grades (Methocel K4, K15 and K100M) were used to obtain different release rates of the drug from the extended-release layer, ketoprofen and praziquantel were used as slightly soluble model drugs. The in vitro dissolution tests of the prepared double-layer systems, showed the desired biphasic behaviour: the drug contained in the fast releasing layer dissolved within the first 15 min, while the drug contained in the prolonged-release layer was released at different times, depending on the formulation of the hydrophilic matrix. In particular, an increase in the percentage and viscosity grade of HPMC, in the extended release layer, leads to a decrease in the drug delivery rate and produces a wide range of different release rates from only a few hours up to 24 h. PMID- 10477328 TI - Preparation of alginate gel beads containing chitosan nicotinic acid salt and the functions. AB - Calcium-induced alginate gel beads containing chitosan salt (Alg-CS) was prepared using nicotinic acid (NA), a drug for hyperlipidemia, and investigated its two functions in gastrointestinal tract, (a) NA release from Alg-CS, (b) uptake of bile acids into Alg-CS. The amount of NA incorporated in Alg-CS increased according to increment of CS content. NA was rapidly released from Alg-CS in diluted HCl solution (pH 1.2) or physiological saline without disintegration of the beads. When Alg-CS was placed in bile acid solution it took bile acid into itself. About 80% of taurocholic acid dissolved in the medium was taken into Alg CS. According to increment of bile acid concentration, the uptake amount increased and an approximately linear relationship existed among them. PMID- 10477309 TI - Bioenergetics of the Archaea. AB - In the late 1970s, on the basis of rRNA phylogeny, Archaea (archaebacteria) was identified as a distinct domain of life besides Bacteria (eubacteria) and Eucarya. Though forming a separate domain, Archaea display an enormous diversity of lifestyles and metabolic capabilities. Many archaeal species are adapted to extreme environments with respect to salinity, temperatures around the boiling point of water, and/or extremely alkaline or acidic pH. This has posed the challenge of studying the molecular and mechanistic bases on which these organisms can cope with such adverse conditions. This review considers our cumulative knowledge on archaeal mechanisms of primary energy conservation, in relationship to those of bacteria and eucarya. Although the universal principle of chemiosmotic energy conservation also holds for Archaea, distinct features have been discovered with respect to novel ion-transducing, membrane-residing protein complexes and the use of novel cofactors in bioenergetics of methanogenesis. From aerobically respiring Archaea, unusual electron-transporting supercomplexes could be isolated and functionally resolved, and a proposal on the organization of archaeal electron transport chains has been presented. The unique functions of archaeal rhodopsins as sensory systems and as proton or chloride pumps have been elucidated on the basis of recent structural information on the atomic scale. Whereas components of methanogenesis and of phototrophic energy transduction in halobacteria appear to be unique to Archaea, respiratory complexes and the ATP synthase exhibit some chimeric features with respect to their evolutionary origin. Nevertheless, archaeal ATP synthases are to be considered distinct members of this family of secondary energy transducers. A major challenge to future investigations is the development of archaeal genetic transformation systems, in order to gain access to the regulation of bioenergetic systems and to overproducers of archaeal membrane proteins as a prerequisite for their crystallization. PMID- 10477329 TI - Densitometric thin layer chromatographic analysis of tretinoin and erythromycin in lotions for topical use in acne treatment. AB - A TLC-method was developed to analyse tretinoin and erythromycin in a lotion in the presence of several excipients. Erythromycin was separated on a silica gel plate and a mobile phase with dichloromethane, methanol and ammonia 25% (60:6:1 (v/v/v)), tretinoin on a C(18) RP plate with acetonitrile and water (50:25 (v/v)) as mobile phase, adding 1 ml acetic acid for the separation of the excipients and erythromycin. The derivatization for both was done with a dipping reagent, consisting of anisaldehyde, sulphuric acid and acetic acid (respectively 1, 2 and 10% (v/v/v)) and dissolved in chloroform/alcohol 94% v/v (60:30 (v/v)) for erythromycin and alcohol 94%/water (50:40 (v/v)) for tretinoin. These TLC-systems were quantitatively evaluated in terms of stability of the colour, precision, accuracy and calibration, proving the utility in the analysis of the lotion. PMID- 10477330 TI - A calorimetric investigation into the interaction between paracetamol and polyethlene glycol 4000 in physical mixes and solid dispersions. AB - The solubility, heat of solution and dissolution rate of paracetamol and polyethyelene glycol 4000 (PEG 4000) systems have been studied in order to clarify the nature of the interaction between the two components during dissolution of solid dispersions. The logarithmic solubility of paracetamol demonstrated a non-linear increase with concentration of PEG 4000, while linear relationships between heat of solution in water and concentration were seen for both individual components. However, the heat of solution of paracetamol was found to decrease with increasing concentrations of PEG 4000. Similarly, the heats of solution in water of physical mixes and solid dispersions prepared using two manufacturing protocols were found to be lower than the theoretical values calculated from those corresponding to the individual components. Drug release studies showed a marked increase in paracetamol dissolution rate when prepared as a solid dispersion, with behaviour consistent with carrier controlled dissolution observed at low drug contents which was ascribed to enhanced dissolution of the drug into the diffusion layer of the PEG 4000. The implications of the understanding of this mechanism for the choice of carrier and manufacturing protocol for solid dispersion products is discussed. PMID- 10477331 TI - Aqueous ethyl cellulose dispersion containing plasticizers of different water solubility and hydroxypropyl methyl-cellulose as coating material for diffusion pellets II: properties of sprayed films. AB - This study investigates the properties of sprayed films prepared from aqueous ethyl cellulose dispersions (ECD) containing hydroxypropyl methylcellulose (HPMC) and plasticizers of different water solubility in order to clarify the drug release mechanisms of pellets coated with the respective material. It is of special interest to measure the migration of the water soluble components as well as the physical properties of the swollen ethyl cellulose film. Swelling experiments with sprayed films in 0.1 N-HCl at 37 degrees C show that fairly water soluble plasticizers and the pore forming agent (HPMC) migrated rapidly and almost completely out of the films. The water insoluble plasticizers remain predominantly in the film and the migration rate of HPMC is reduced in a release medium of high ionic strength. The glass transition temperature (T(g)) and the softening temperature (T(s)) of these films after swelling are dependent on the water solubility of the plasticizer. The T(g) of ECD films plasticized with triethyl citrate is above the swelling temperature of 37 degrees C after migration of the plasticizer, transforming the polymer in the glassy state. In contrast, dibutyl phthalate-containing ECD films demonstrate a T(g) below the swelling temperature, leaving the polymer in the rubbery state. The mechanical properties of dry and wet films are studied as a function of the state of curing of the films and of the swelling temperature. On contact with water, a pronounced shrinkage of ECD/HPMC films plasticized with water insoluble plasticizers is observed. All these results are used to explain the different drug release mechanisms of the coated pellets and to enable the prediction and optimization of drug release-rates from coated pellets. PMID- 10477332 TI - Formation and structure of stably dispersed particles composed of retinal with dipalmitoylphosphatidylcholine: coexistence of emulsion particles with bilayer vesicles. AB - In order to develop an intravenous formulation of all-trans-retinal (vitamin A aldehyde, VAA) for the treatment of night blindness, VAA and dipalmitoylphosphatidylcholine (DPPC) were sonicated and the dispersions in the VAA mole fraction range of 0.1-0.7 were stable at room temperature for 3 days. In order to clarify the dispersal mechanism, the dispersed particles were characterized and the interaction between VAA and DPPC was investigated using several physicochemical techniques. Dynamic light scattering measurements showed that the diameter of the dispersed particles was 50-70 nm. A limited amount of VAA is incorporated into DPPC bilayer membranes (approximately 5 mole%). The trapped aqueous volume inside the particles was determined fluorometrically using the aqueous space marker calcein and the volume in the VAA/DPPC particles was decreased remarkably with the addition of VAA into small unilamellar vesicles of DPPC. The decline in the fraction of vesicular particles was also confirmed by fluorescence quenching of N-dansylhexadecylamine in the DPPC membrane by the addition of the quencher CuSO(4). These results indicate that the excess VAA separated from the DPPC bilayers is stabilized as emulsion particles by the DPPC surface monolayer. The monolayer-bilayer equilibrium of VAA/DPPC mixtures was estimated by measurement of spreading and collapse pressures. The results showed that the coexistence of emulsion particles (surface monolayer of DPPC+core of VAA) with vesicular particles (bilayer) was critically important for the formation of the stably dispersed particles of the lipid mixture. PMID- 10477333 TI - Evaluation of cytotoxicity and absorption enhancing effects of melittin - a novel absorption enhancer. AB - Melittin is the major active ingredient in bee venom and has been widely studied for its membrane-fusion property. We have explored the possibility of determining a concentration range of melittin where it is relatively safe to be used as an adsorption enhancer. Melittin's potential use as an adsorption enhancer for mannitol was determined using Caco-2 cells as the model epithelial membrane. The results indicated that at concentrations below 2.4 (M melittin is safe. Using a melittin concentration of 1.5 (M there was 3.5 times increased transepithelial transport of mannitol across Caco-2 cell monolayers. PMID- 10477334 TI - Lymphocyte DNA damage in cigarette factory workers measured by the Comet assay. AB - To investigate whether there were separate and combined effects of occupational exposure to tobacco dust and smoking on lymphocyte DNA damage, 148 workers from a cigarette manufacturing factory (107 occupationally exposed to tobacco dust from the production department and 41 unexposed controls who were managerial workers) were included in the study. The Tail Moment (TM) of Comet assay was used to measure DNA damage. The two groups had similar mean age, mean duration of work and smoking prevalence. The exposed workers had a larger TM than that of the controls (mean+/-S.D.: 43.43+/-13. 77 vs. 38.89+/-8.98, p<0.05). Smokers had significantly larger TM than non-smokers (47.25+/-14.02 vs. 38.90+/-10.75, p<0.001). Analysis of variance after adjustment for age and gender showed that occupational exposure and smoking had a significant and independent effect on Tail Moment (p=0.025 and p=0.002, respectively) and there was a significant positive two way interaction between the two factors (p=0.019). Age and gender had no significant effect on TM. The present study suggests that smoking and tobacco dust exposure can induce lymphocyte DNA damage and there is a synergistic effect of tobacco dust exposure and smoking on DNA damage. PMID- 10477336 TI - Mutagenic and genotoxic effects of theophylline and theobromine in Salmonella assay and in vivo sister chromatid exchanges in bone marrow cells of mice. AB - The mutagenic and genotoxic effects of two methylxanthines, theophylline (TH) and theobromine (TB), were assessed in the Ames mutagenicity assay (in strains TA97a, TA100, TA102 and TA104) and in vivo sister chromatid exchanges (SCEs) in bone marrow cells of mice. These are the two most commonly used nervous system stimulators throughout the world. TH is used in the long-term treatment of asthma. Bacterial mutagenicity assay showed very weak mutagenic effects of both drugs in Salmonella strains TA102 and TA104 only in certain concentrations when S9 was added to it. No mutagenic effects were observed in any other strains used in this assay either with or without metabolic activation. But results of in vivo SCE assay indicate that these two drugs can induce significant SCE in bone marrow cells of mice. PMID- 10477335 TI - Mutagenicity of commercial Monascus fermentation products and the role of citrinin contamination. AB - Pigments produced as secondary metabolites by various isolates of moulds belonging to the genus Monascus have been used traditionally as colorants in Oriental food. Modern food industry has rediscovered these moulds as promising source for natural colorants. However, recent studies evidence that one of the secondary metabolites produced by Monascus is identical in structure to the mycotoxin citrinin. Thus, a sensitive HPLC method was developed to analyse these food colorants for contamination with citrinin. The mycotoxin could be detected in all the commercial Monascus samples at concentrations varying between 0.2 to 17.1 microg/g. In addition, the mutagenicity of commercial Monascus samples applying Salmonella-microsome assay and Salmonella-hepatocyte-assay was investigated and compared to the results obtained with citrinin. Citrinin and two Monascus extracts induced a positive dose depending mutagenic response in the Salmonella-hepatocyte-assay applying strain TA-98. However, no mutagenicity could be detected in the Salmonella-microsome assay, neither with nor without S9-mix, for citrinin and Monascus extracts, applying TA-98, TA-100, TA-1535, TA-1538 and TA-97. These findings provide further evidence that citrinin requires complex cellular biotransformation to exert mutagenicity. PMID- 10477337 TI - Cytotoxic and genotoxic effects of energetic compounds on bacterial and mammalian cells in vitro. AB - The mutagenicity and toxicity of energetic compounds such as 2,4, 6 trinitrotoluene (TNT), 1,3,5-trinitrobenzene (TNB), hexahydro-1,3, 5-trinitro 1,3,5-triazine (RDX) and octahydro-1,3,5,7-tetranitro-1,3, 5,7-tetrazocine (HMX), and of amino/nitro derivatives of toluene were investigated in vitro. Mutagenicity was evaluated with the Salmonella fluctuation test (FT) and the V79 Chinese hamster lung cell mutagenicity assay. Cytotoxicity was evaluated using V79 and TK6 human lymphoblastic cells. For the TK6 and V79 assays, TNB and 2, 4,6 triaminotoluene were more toxic than TNT, whereas RDX and HMX were without effect at their maximal aqueous solubility limits. The primary TNT metabolites (2-amino 4,6-dinitrotoluene, 4-amino-2, 6-dinitrotoluene, 2,4-diamino-6-nitrotoluene and 2, 6-diamino-4-nitrotoluene) were generally less cytotoxic than the parent compound. The FT results indicated that TNB, TNT and all the tested primary TNT metabolites were mutagenic. Except for the cases of 4-amino-2,6-dinitrotoluene and 2,4-diamino-6-nitrotoluene in the TA98 strain, addition of rat liver S9 resulted in either no effect, or decreased activity. None of the tested compounds were mutagenic for the V79 mammalian cells with or without S9 metabolic activation. Thus, the FT assay was more sensitive to the genotoxic effects of energetic compounds than was the V79 test, suggesting that the FT might be a better screening tool for the presence of these explosives. The lack of mutagenicity of pure substances for V79 cells under the conditions used in this study does not preclude that genotoxicity could actually exist in other mammalian cells. In view of earlier reports and this study, mutagenicity testing of environmental samples should be considered as part of the hazard assessment of sites contaminated by TNT and related products. PMID- 10477338 TI - Geographical variations in the concentration of biliary free fatty acids with anti-mutagenic action. AB - The concentrations and compositions of free fatty acids (FFAs) in human bile, especially of inhibitory free fatty acids (IFFAs), were analyzed in terms of anti mutagenic effects in relation to the mutagenic activity of bile. Bile samples were collected from patients with cholelithiasis residing in either Niigata or Kochi prefectures of Japan, regions characterized as the highest and lowest risk areas for gallbladder cancer (GBC), respectively. Biliary FFAs and IFFAs were analyzed by high-performance liquid chromatography and mutagenicity was examined in by the Ames test (TA98+S9mix) after blue rayon treatment. There was a tendency for higher biliary FFA and IFFA concentrations in the Kochi subjects, but the proportion of IFFA to the total FFA concentration did not differ between the two areas. There was an inverse correlation between the concentrations of IFFAs and the numbers of revertant colonies in both Niigata and Kochi subjects. However, at a dose of 591 micromol/l, (calculated based on the average amount of IFFAs absorbed in blue rayon) IFFAs did not exhibit anti-mutagenic actions in the blue rayon extracts. Within this range, more positive samples were seen in Niigata than in Kochi, suggesting the presence of more active mutagen(s) in Niigata samples. PMID- 10477339 TI - Illumination of human keratinocytes in the presence of the sunscreen ingredient Padimate-O and through an SPF-15 sunscreen reduces direct photodamage to DNA but increases strand breaks. AB - On illumination with simulated sunlight, the UVB-absorbing sunscreen chemical 2 ethylhexyl-4-dimethylaminobenzoate (Padimate-O) generates excited species which inflict non-ligatable strand breaks on DNA in vitro and it also becomes mutagenic to yeast in vivo. Padimate-O is known to penetrate human skin but its effects on human cells are not clear. Here, we first simulate the sunlight which penetrates human skin and use it to illuminate human keratinocytes. The DNA damage observed in terms of UV-endonuclease-sensitive sites (ESS) and direct strand breaks per kilobase (kb) of DNA per joule per square metre agrees well with that predicted from action spectra based on monochromatic light. Using plasmid DNA in vitro, we find a very similar pattern of results. Next, we simulate the spectrum that results when the incident light is first attenuated by a film of sunscreen (SPF 15; 2 mg/cm(2)) containing benzophenone-3 (a UVA absorber), octyl methoxycinnamate (a UVB absorber), and Padimate-O. If the sunscreen is not in contact with keratinocytes it reduces direct DNA damage from sunlight (ESS). However, any Padimate-O in contact with the cells substantially increases indirect damage (strand breaks) even though the film of sunscreen reduces direct photodamage. We estimate that applying an SPF-15 sunscreen which contains Padimate-O to human skin followed by exposure to only 5 minimum erythemal doses (MED) of sunlight could, while suppressing the formation of ESS, increase strand breaks in cells under the epidermis by at least 75-fold compared to exposure to 1 MED in the absence of sunscreen. PMID- 10477340 TI - DNA damage detected by the comet assay in the white blood cells of workers in a wooden furniture plant. AB - The study was aimed at the assessment of genotoxic effects in workers of a wooden furniture manufacture, based on the level of DNA damage in white blood cells (WBC). The alkaline single cell gel electrophoresis assay (known as the comet assay) in individual cells was adapted for detecting damaged DNA in WBC. The level of DNA damage was determined as the percentage of cells with comets. It was assessed in cells before and after incubation in RPMI 1640 medium and CO(2) at 37 degrees C for 1 h to repair DNA breaks. Thirty-five woodworkers and 41 control subjects were studied. In the woodworkers, significantly more cells with DNA damage (21.5%) were observed than in the control persons (9.7%). A slight but significant decrease in the level of DNA damage was found in the WBC of woodworkers after incubation (17.2%). Significantly higher levels of damaged DNA was observed in woodworkers who either smoked (22.1%) or did not smoke cigarettes (20.8%) than in smokers (13.2%) and non-smokers (7.0%) from the control group. After incubation, a slight decrease in the level of DNA damage was found in both smoking and non-smoking woodworkers compared to the respective subjects in the control group. The increased levels of DNA damage observed in the woodworkers could be associated with the occupational exposure to wood dust in the furniture manufacture. PMID- 10477341 TI - A simple and rapid method for analyzing the Monascus pigment-mediated degradation of mutagenic 3-hydroxyamino-1-methyl-5H-pyrido[4,3-b]indole by in-capillary micellar electrokinetic chromatography. AB - A simple and rapid method is described for analyzing the Monascus pigment mediated degradation of 3-hydroxyamino-1-methyl-5H-pyrido[4, 3-b]indole (Trp-P 2(NHOH)). We used the in-capillary micellar electrokinetic chromatography (MEKC). During the electrophoresis, the mutagen and the pigment, due to their different migration velocities, mix for a certain period of time to interact, and then they are separated and quantified. Using this technique, we were able to demonstrate that Trp-P-2(NHOH) is degraded by the pigment. The degradation was pigment-dose dependent, and because the pigment was recovered unchanged, it was deduced that the pigment acted catalytically for the degradation. The entire MEKC procedure takes 8 min. PMID- 10477342 TI - Mutagenesis of a single AT basepair in mice transgenic for PhiX174 am3, cs70. II. Brain. AB - Most cell divisions in the mouse brain have ceased by 14 days after birth. Therefore, spontaneous mutations that occur in brain cells can be assumed to be fixed by replication during brain development. Spontaneous and ethylnitrosourea (ENU)-induced reverse mutations at a single AT base pair were measured in brain tissue by using mice transgenic for PhiX174 am3, cs70. The line (am54) has 50 PhiX genomes per haploid genome integrated in a tandem array and is maintained by random breeding on a C57BL/6 background. For mutagenesis studies, homozygous am54 males were mated to non-transgenic C57BL6/J females. Four-day old offspring from this cross were treated with 50 mg/kg ENU and were euthanized at 68-80 days of age. The ENU-treated animals had a significantly higher frequency of am3 revertants in brain than did concurrent controls. In a second experiment, hemizygous male offspring (85 to 94 days old) were treated with 150 mg/kg ENU and euthanized at various post-injection intervals (3, 10 and 110 days). The revertant frequencies 3 and 10 days after treatment were not significantly different from control values. At the 110 days post-injection interval, however, the average revertant frequency in the treated group was significantly lower than controls. In a second study animals were euthanized 3, 10 and 74 days after treatment. Two groups (3 and 74 days post-injection) also showed a significant decrease in the revertant frequency as compared to controls. Additional sets of adult animals were treated with 50 and 150 mg/kg ENU and were euthanized 195 to 201 days after treatment. The average revertant frequency of the animals that were treated with 50 or 150 mg/kg ENU was not significantly different from the control value. Thus, although an increase in mutant frequency is detected in the PhiX174 system when neonatal mice are treated with ENU, treatment of mature mice with ENU did not result in an increase in the mutant frequency. Moreover, under certain conditions, ENU-produced a significantly lower mutant frequency than was observed in the control animals. This decrease in the revertant frequency among the treated animals was likely due to selective killing of cells with a higher spontaneous revertant frequency than cells with lower frequency. PMID- 10477343 TI - Pluripotent stem cells of the mouse as a potential in vitro model for mammalian germ cells. Sister chromatid exchanges induced by MMC and ENU in undifferentiated cell lines compared to differentiated cell lines. AB - We tried to develop an in-vitro test system which could serve as a model for mammalian germ cells in vivo. Two pluripotent cell types were used, because they express some germ cell specific immunological and biochemical markers: (1) Embryonal carcinoma cells (ECC) of the line P19 had been isolated from a teratocarcinoma of murine primordial germ cells (PGC). (2) Embryonal stem cells (ESC) are obtained from the inner cell mass of mouse blastocysts. Sister chromatid exchanges (SCE) induced by mitomycin C and ethylnitrosourea (ENU) were analysed in the two undifferentiated cell lines, ECC and ESC, to detect differences in their sensitivity compared with differentiated cell lines of the mouse. Neither of the model cell lines have shown a greater sensibility after exposure to MMC and ENU. In contrary, the carcinoma cell line was able to tolerate higher concentrations of these genotoxicants. Therefore, SCE analysis in the ECC and ESC lines used does not provide a suitable model for genotoxicity testing on mammalian germ cells. PMID- 10477344 TI - Comparative in vitro and in vivo assessment of genotoxic effects of etoposide and chlorothalonil by the comet assay. AB - The alkaline single cell gel electrophoresis (comet) assay was used to assess in vitro and in vivo genotoxicity of etoposide, a topoisomerase II inhibitor known to induce DNA strand breaks, and chlorothalonil, a fungicide widely used in agriculture. For in vivo studies, rats were sacrificed at various times after treatment and the induction of DNA strand breaks was assessed in whole blood, bone marrow, thymus, liver, kidney cortex and in the distal part of the intestine. One hour after injection, etoposide induced DNA damage in all organs studied except kidney, especially in bone marrow, thymus (presence of HDC) and whole blood. As observed during in vitro comet assay on Chinese hamster ovary (CHO) cells, dose- and time-dependent DNA effects occurred in vivo with a complete disappearance of damage 24 h after administration. Even though apoptotic cells were detected in vitro 48 h after cell exposure to etoposide, such a result was not found in vivo. After chlorothalonil treatment, no DNA strand breaks were observed in rat organs whereas a clear dose-related DNA damage was observed in vitro. The discrepancy between in vivo and in vitro models could be explained by metabolic and mechanistic reasons. Our results show that the in vivo comet assay is able to detect the target organs of etoposide and suggest that chlorothalonil is devoid of appreciable in vivo genotoxic activity under the protocol used. PMID- 10477345 TI - Protective effect of the yeast glucomannan against cyclophosphamide-induced mutagenicity. AB - Glucomannan (GM) isolated from Candida utilis with molecular weight 30 kDa was administered either intraperitoneally or orally prior to cyclophosphamide (CP) injection and its effect on the frequency of micronuclei was evaluated in polychromatic erythrocytes of mouse bone marrow. GM administration by either route decreased significantly (p<0.002) the clastogenic effect of CP. The protective effect was concentration-dependent, with a higher decrease achieved by 200 mg/kg than by 100 mg/kg b. wt. (body weight). The fact that GM was effective also at oral administration is indicative of the passage of GM molecules through the wall of the gastrointestinal tract. The important characteristics of GM isolated from C. utilis, such as good water solubility, relatively small molecular weight (30 kDa), and antimutagenic effect exerted also at oral administration, appear to be promising features for its prospective use as a natural protective agent. PMID- 10477346 TI - Molecular analysis of mutations induced by a benzene metabolite, p-benzoquinone, in mouse cells using a novel shuttle vector plasmid. AB - Human population has been continually exposed to benzene which is present in our environment as an essential component of petroleum. p-Benzoquinone (p-BQ) is one of the benzene metabolites and is thought to be an ultimate toxic or carcinogenic substance. For molecular analysis of carcinogen-induced mutations in mouse cells, we constructed a new shuttle vector plasmid pNY200 that has supF gene as a target of the mutations and replicates in mouse and in Escherichia coli cells. In p-BQ treated pNY200 propagated in mouse cells, base substitutions were induced predominantly at G:C sites, and the major mutation was G:C-->A:T transition. Many tandem base substitutions were also induced at CC:GG sequences. By a postlabeling analysis and a polymerase stop assay, we confirmed that p-BQ adducts formed in DNA and mutation sites roughly correspond to the sites where the adducts were formed. Comparing data of pNY200 in mouse cells with those of the similar shuttle vector plasmid pMY189 in human cells should be important for extrapolation of data from mouse to human, because carcinogenicity of chemicals is tested in mice. PMID- 10477347 TI - Participation of the pyrazine cation radical in the formation of mutagens in the reaction of glucose/glycine/creatinine. AB - Two classes of the free radical Maillard intermediates, the pyrazine cation radical and the carbon-centered radicals, are detected in the reaction of Glc (glucose)/Gly (glycine) by electron spin resonance and spin trapping technique. Profile of the generation of the pyrazine cation radical in the reaction with different ratios of the reactants was found to be similar to that of the formation of mutagens in the subsequent reaction with creatinine. By contrast, profile of the generation of the carbon-centered radicals was not consistent with that of the mutagen formation. Thiol antioxidants and unsaturated fatty acids (or their esters) effectively scavenged the pyrazine cation radical generated in the reaction of Glc/Gly, and inhibited the formation of the mutagens in the reaction of Glc/Gly and creatinine. Ethanol, a sulfide and a saturated fatty acid were not effective to scavenge the pyrazine cation radical and did not inhibit the mutagen formation. The pyrazine cation radical rather than the carbon-centered radicals may play an important role in the mutagen formation. Thiol antioxidants and unsaturated fatty acids can be evaluated as inhibitors of the pyrazine cation radical-derived formation of the mutagens. PMID- 10477348 TI - Male-mediated dominant lethal mutations in mice following prooxidant treatment. AB - This study's primary aim is to examine if prooxidant treatment has the propensity to induce dominant lethal (DL) type mutations in a randomly bred closed colony of CFT-Swiss mice. Initially, graded doses of both organic hydroperoxides viz., t butyl hydroperoxide (tbHP), and cumene hydroperoxide (cHP) were administered (i.p.) to adult males and the mortality data was analysed to determine the LD(50) values. cHP was relatively more toxic compared to tbHP. The computed LD(50) values were 1500 and 3000 micromol (kg body weight)(-1) for cHP and tbHP, respectively. Subsequently, adult males were administered (i.p.) with 1/10 LD(50) doses of hydroperoxide (HP) (tbHP--30 micromol (100 g body weight)(-1) and cHP - 15 micromol (100 g body weight)(-1)) on 5 consecutive days and were mated with virgin females for a period of 5 weeks to characterise the male-mediated DL mutations. Male-based analysis of the three major variables viz., implantations, live embryos and dead implants (DI) were carried out to assess the DL-type response induction. While tbHP induced significant increases (2- to 5-fold) in the incidence of DI during the first 4 weeks, cHP induced a marginal increase only during the first week. These results suggest that prooxidants induce DL-type effect only in specific post-meiotic stages of spermatogenesis and stress the need to further investigate the implications of chronic oxidative stress on the male reproductive system. PMID- 10477349 TI - Cell cycle specificity of cytogenetic damage induced by 3,4-epoxy-1- butene. AB - 3,4-epoxy-1-butene (EB), a primary metabolite of butadiene, is a direct-acting "S dependent" genotoxicant that can induce sister chromatid exchanges (SCEs) and chromosome aberrations (CAs) in cycling cells in vitro. However, EB is almost inactive when splenic or peripheral blood lymphocytes are exposed at the G(0) stage of the cell cycle. To investigate whether repair of DNA lesions is responsible for the lack of cytogenetic responses seen after G(0) treatments, we used cytosine arabinoside (ara-C) to inhibit DNA polymerization during DNA repair. If enough repairable lesions are present, double-strand breaks should accumulate and form chromosome-type ("S-independent") deletions and exchanges. This is exactly what occurred. EB induced chromosome deletions and dicentrics at the first division following treatment, when the EB exposure was followed by ara C. Without ara-C treatment, there was no induction of CAs. These experiments indicate that the relatively low levels of damage induced by EB in G(0) lymphocytes are removed by DNA repair prior to DNA synthesis and thus, before the production of SCEs or chromatid-type aberrations. PMID- 10477350 TI - Study of a rat skin in vivo micronucleus test: data generated by mitomycin C and methyl methanesulfonate. AB - We have developed an in vivo micronucleus (MN) test that uses rat skin as the target organ. Sample preparation involves cold-treating the epidermis with trypsin, peeling it off with a fine forceps, treating it in hypotonic solution, and staining it with acridine orange (A.O.). We evaluated the assay using mitomycin C (MMC) and methyl methanesulfonate (MMS) as model clastogens, applying them as single and repeat treatments. Both chemicals induced a significant, dose dependent increase in MN frequency in basal cells. One treatment per day for 3 days was optimal for MN induction. PMID- 10477351 TI - Genotoxic effects of triphenyltin acetate and triphenyltin hydroxide on mammalian cells in vitro and in vivo. AB - Two organotin pesticides, triphenyltin acetate (TPTA) and triphenyltin hydroxide (TPTH), were evaluated for their ability to induce micronuclei (MN) and sister chromatid exchange (SCE) in vitro using cultured Chinese hamster ovary (CHO) cells and in vivo BALB/c mouse erythrocytes. Both pesticides induced a dose dependent increase but only TPTH induced a significant increase in MN at the highest dose (150 ng/ml) tested in CHO cells. With adding S9 microsomal fractions, both pesticides induced a meaningful MN induction at 150 ng/ml and a dose-dependent significant increase in SCE. In vivo MN induction in erythrocytes was conducted by treating BALB/c mice orally or intraperitoneally with these pesticides either in a single or triple treatments. Oral gavage (p.o.) of TPTA resulted in a dose-related significant increase of MN induction in peripheral blood and of TPTH induced a significant increase in micronucleated reticulocyte (MNRETs) only in a single treatment. Intraperitoneal administration of TPTA or TPTH, however, resulted in meaningless random increases in MN though these increases might be attributable to toxic effects. The MNRETs levels in the treatment with both pesticides were independent to the sampling time. This study demonstrated that TPTA and TPTH was potential chromosome mutagens. PMID- 10477352 TI - Genotoxicity studies on the azo dye Direct Red 2 using the in vivo mouse bone marrow micronucleus test. AB - The clastogenicity of the azo dye Direct Red 2 (DR2) has been investigated using the murine bone marrow micronucleus assay. A potent dose-dependent response was observed following oral gavage of DR2 up to 4 mg/kg, after which significant toxicity to the erythroid compartment was observed. The route of administration had a significant effect on the frequency of micronucleus formation: intraperitoneal injection was approximately two-fold less clastogenic than the equivalent dose delivered orally (p<0.05). The requirement for activation of DR2 by intestinal microflora was indicated by the fact that mice given acid-treated water prior to administration of DR2 showed a significant reduction (40%; p<0.001) in micronucleated polychromatic erythrocyte formation. The implications of these findings for the health and safety of occupationally exposed workers are discussed. PMID- 10477353 TI - Enhancement of bleomycin-induced micronucleus formation in V79 cells as a rapid and sensitive screen for non-covalent DNA-binding compounds. AB - Non-covalent drug/DNA interactions are difficult to study and because of this, the significance of such interactions from a safety standpoint and their contribution to positive genetic toxicology test findings is poorly understood. It is shown in the present study that such interactions may be detected and quantified in Chinese hamster V79 cells by an adaptation of the bleomycin amplification assay. This assay measures the ability of a test compound to enhance the DNA damaging activity of the antibiotic bleomycin using micronucleus formation as an endpoint. Results are presented examining the bleomycin amplification activity of known intercalating agents, groove-binding agents and other structurally diverse classes of compounds for which intercalative status has not been reported. The assay reveals a strong and predictable SAR for amplification activity based on number and orientation of aromatic rings. Moreover, excellent correlations are observed between DNA binding (viscometric analyses) and DNA amplification in V79 cells for a series of seven experimental compounds. The assay is shown to be useful in understanding the genotoxicity of marketed antihistamines and to help explain genetic toxicology findings observed in a series of novel pharmaceutical entities. It is proposed that assessment of bleomycin amplification activity of novel compounds in early genotoxicity prescreening may provide important information upon which to base synthesis of compounds with minimal or no genotoxic liability. PMID- 10477354 TI - Neoplastic potential of rat tracheal epithelial cell lines induced by 1 nitropyrene and dibenzo(a,i)pyrene. AB - Our previous study showed that both 1-nitropyrene (1-NP) and dibenzo(a,i)pyrene (DBP) induced enhanced growth variants (EGVs) in primary cultures of rat tracheal epithelial (RTE) cells exposed in vivo. Cell lines were established from some of the EGVs. Further studies, using anchorage-independent growth in soft agar and tumorigenicity in athymic nude mice, were performed to determine the neoplastic potential of EGVs induced by 1-NP and DBP. Results show that three of five from DBP- and five of five from 1-NP-induced cell lines displayed anchorage independent growth. The colony forming efficiency (CFE) from DBP-induced cell lines was 0.067 per thousand and CFE from 1-NP-induced cell lines was 0.151 per thousand. There is a significant difference between the two CFEs (mu = 12.08, P<0. 01). Two of five DBP- and five of five 1-NP-induced cell lines produced squamous cell carcinomas (SCC) in nude mice. The rate of tumorigenicity counted by injected sites was 20% (6/30) for DBP-induced cell lines and 57% (17/30) for 1 NP-induced cell lines. There is a significant difference between the results of tumorigenicity from the cell lines induced by the two different compounds (chi(2)=8.53, P<0.01). Neither of the two cell lines from spontaneously developed foci grew in soft agar or produced SCC in nude mice. It seems that the neoplastic potential of transformed RTE cells induced by 1-NP was higher than that of DBP. PMID- 10477355 TI - Effect of blood storage on radiation-induced micronuclei in human lymphocytes. AB - To evaluate the effect of blood storage on the yield of micronuclei (MN) in both irradiated (in vivo and ex vivo) and unirradiated peripheral blood lymphocytes (PBL), we applied the MN assay in cytokinesis-blocked (CB) PBL obtained from healthy subjects (n=11), and from cancer patients (n=10) who were undergoing fractionated partial-body radiotherapy (xRT). The heparinized blood samples were exposed to 137Cs-irradiation (0 Gy or 2 Gy) immediately after blood collection and were stored upright in test tubes either at room temperature (22 degrees C) or in the refrigerator (5 degrees C). Duplicate whole blood cultures from each sample were set up at 0 h, 96 h, and 120 h after ex vivo irradiation. Giemsa (10%) stained slides were prepared from each culture. MN yield was determined per 1000 binucleated cells. As compared to that obtained from the corresponding fresh blood samples, we found that (1) the 22 degrees C blood storage temperature did not affect MN yields in PBL of either healthy subjects or cancer patients up to 96 h, either with or without ex vivo irradiation; and (2) while blood samples were stored at 5 degrees C, the MN yield increased significantly in PBL of healthy subjects (with or without ex vivo irradiation) at 120 h, and in cancer patients (with ex vivo irradiation) at 96 h and 120 h. Since handling of the blood sample is important for CBMN assay during shipment or in the laboratory, our findings showed that blood storage at 22 degrees C or at 5 degrees C up to 96 h appeared to provide insignificant variations of the MN results as compared to fresh blood samples. However, the 96 h of blood storage at 5 degrees C elevated the MN frequency in ex vivo irradiated PBL of cancer patients who were undergoing xRT. PMID- 10477356 TI - Ames assays and unscheduled DNA synthesis assays on 2, 4-dichlorophenoxyacetic acid and its derivatives. AB - 2,4-dichlorophenoxyacetic acid and several of its derivatives (collectively known as 2,4-D) are herbicides used to control a wide variety of broadleaf and woody plants. The genetic toxicity in vitro of 2,4-D and seven of its salts and esters were examined by employing gene mutation in bacteria (Ames test) and induction of DNA damage and repair in rat hepatocytes. In addition, an in vivo unscheduled DNA synthesis (UDS) assay was performed on 2,4-D. There were no indications of genotoxic potential for 2,4-D acid, or any of its derivatives, in these assays. These results are consistent with the reported lack of carcinogenic potential for 2,4-D in both mice and rats. PMID- 10477357 TI - Evaluation of the genotoxicity of 2,4-dichlorophenoxyacetic acid and its derivatives in mammalian cell cultures. AB - 2,4-dichlorophenoxyacetic acid and its derivatives (collectively known as 2,4-D) are herbicides used to control a wide variety of broadleaf and woody plants. The genetic toxicity of an ester (2,4-D 2-butoxyethylester) and two salts (2,4-D isopropylamine and 2,4-D triisopropanolamine) was investigated in cultured mammalian cells. The end points used were the induction of chromosomal aberrations in primary cultures of rat lymphocytes and forward mutations at the HGPRT locus of Chinese hamster ovary cells. There was no evidence of genotoxicity for the test materials in the experimental systems used. These results were consistent with the general lack of genotoxic potential for 2,4-D in a number of other test systems. PMID- 10477358 TI - In vivo micronucleus assays on 2,4-dichlorophenoxyacetic acid and its derivatives. AB - The potential for 2,4-D and seven of its salts and esters to induce cytogenetic abnormalities in mammalian cells in vivo was investigated in the mouse bone marrow micronucleus test. All the test materials were administered to male and female mice by oral gavage and the frequencies of micronucleated polychromatic erythrocytes (MN-PCE) in the bone marrow were determined at intervals of 24, 48 and 72 h following dosing. There were no significant increases in the incidence of MN-PCE in the treated mice at any of the bone marrow sampling times. These results are consistent with the reported lack of in vitro genetic toxicity for these materials in various in vitro genotoxicity assays as well as the absence of carcinogenic potential for 2,4-D in both mice and rats. PMID- 10477359 TI - Antimutagenic effects and possible mechanisms of action of vitamins and related compounds against genotoxic heterocyclic amines from cooked food. AB - Possible antimutagenic activity of 26 vitamins and related compounds - ascorbic acid, beta-carotene, cyanocobalamin, folic acid, nicotinic acid, nicotinamide, pantothenic acid, pyridoxale, pyridoxamine, pyridoxine, retinal, retinol, retinoic acid, retinyl acetate, retinyl palmitate, riboflavin, riboflavin 5' phosphate, flavin adenine dinucleotide (FAD), alpha-tocopherol, alpha-tocopherol acetate, vitamins K(1), K(3), K(4), 1, 4-naphthoquinone, and coenzyme Q(10) - was tested against six heterocyclic amine (HCA) mutagens, i.e., 2-amino-3-methyl imidazo[4, 5-f]quinoline (IQ), 2-amino-3,4-dimethyl-imidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethyl-imidazo[4,5-f]quinoxaline (MeIQx), 2-amino-1-methyl 6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino-6-methyl-dipyrido[1,2-a:3',2' d]imidazole (Glu-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) in the Salmonella/reversion assay using tester strains Salmonella typhimurium TA 98 and TA 100. Retinol, retinal, riboflavin, riboflavin 5'-phosphate, FAD, vitamins K(1), K(3), K(4), 1, 4-naphthoquinone, and coenzyme Q(10) caused a concentration dependent decrease in the mutagenicity of all six mutagens in both tester strains. Quantification of antimutagenic potencies by calculating ID(50)1000; vitamin K(1): 401-740; vitamin K(3) (menadione): 85-590; vitamin K(4): 45-313; 1,4-naphthoquinone: 170-290; coenzyme Q(10): 490-860. In general, there were no major differences between HCAs tested except in part with Trp-P-2 nor between the two tester strains. In enzyme kinetic experiments with Salmonella, retinol, vitamins K(3), and K(4) behaved as competitive inhibitors of IQ induced mutagenesis. However, at the highest concentration of menadione (200 nmol/plate) and of riboflavin 5'-phosphate (2000 nmol/plate), non-competitive inhibition was observed. At other concentrations of riboflavin 5'-phosphate and at all concentrations of FAD, meaningful interpretation of enzyme kinetics were not possible. Reduction of the activity of 7-ethoxy- and 7-methoxyresorufin-O dealkylases with IC(50) values of 2.03-30.8 microM indicated strong inhibition of 1A1 and 1A2 dependent monooxygenases by menadione and retinol. Riboflavin 5' phosphate and FAD were less effective (IC(50): 110-803.7 microM). Nicotinamide adenine-dinucleotidephosphate (NADPH) cytochrome P-450 reductase was not affected by retinoids but stimulated by naphthoquinones and both riboflavin derivatives up to about 50 and 80%, respectively. Again, the mutagenic activity of N-hydroxy-2 amino-3-methyl-imidazo[4,5-f]quinoline (N-OH-IQ) in Salmonella was not suppressed by K-vitamins but marginally reduced by retinol, retinal, and FAD but distinctly by riboflavin 5'-phosphate. In various experiments designed for modulation of the mutagenic response, inhibition of metabolic activation of IQ to N-OH-IQ was found to be the only relevant mechanism of antimutagenesis of menadione while a weak contribution of an other way seemed possible for retinol and FAD. PMID- 10477361 TI - Creatine supplementation and swimming performance. AB - The purpose of this study was to determine if oral creatine (CR) ingestion, compared to a placebo (PL), would enable swimmers to maintain a higher swimming velocity across repeated interval sets over 2 weeks of supplementation. Fourteen female and 18 male university swimmers consumed a PL during a 2-week baseline period. Using a randomized, double-blind design, during the next 2 weeks subjects consumed either CR or PL. Swimming velocity was assessed twice weekly during 6 X 50-m swims and once weekly during 10 X 25-yd swims. There was no effect of CR on the 10 X 25-yd interval sets for men and women and no effect on the 6 X 50-m interval sets for women. In contrast, for men, CR significantly improved mean overall swimming velocity in the 6 X 50-m interval after 2 weeks of supplementation, whereas PL had no effect. Although ineffective in women, CR supplementation apparently enables men to maintain a faster mean overall swimming velocity during repeated swims each lasting about 30 s; however, CR was not effective for men in repeated swims each lasting about 10 - 15 s. PMID- 10477360 TI - The effect of a carbohydrate--arginine supplement on postexercise carbohydrate metabolism. AB - The effect of a carbohydrate-arginine supplement on postexercise muscle glycogen storage was investigated. Twelve well-trained cyclists rode for 2 hr on two separate occasions to deplete their muscle glycogen stores. At 0, 1, 2, and 3 hr after each exercise bout, the subjects ingested either a carbohydrate (CHO) supplement (1 g carbohydrate/kg body weight) or a carbohydrate-arginine (CHO/AA) supplement (1 g carbohydrate/kg body mass and 0.08 g arginine-hydrochloride/kg body weight). No difference in rate of glycogen storage was found between the CHO/AA and CHO treatments, although significance was approached. There were also no differences in plasma glucose, insulin, or blood lactate responses between treatments. Postexercise carbohydrate oxidation during the CHO/AA treatment was significantly reduced compared to the CHO treatment. These results suggest that the addition of arginine to a CHO supplement reduces the rate of CHO oxidation postexercise and therefore may increase the availability of glucose for muscle glycogen storage during recovery. PMID- 10477362 TI - A comparison of the gastric emptying characteristics of selected sports drinks. AB - The purpose of this study was to determine the effects of repeated ingestion of drinks containing varying concentrations of carbohydrate on gastric emptying rate during steady-state exercise. On five separate occasions, 14 subjects cycled for 90 min at an average power output of 151 +/- 2 W. At 15-min intervals, subjects ingested 227 +/- 3 ml of either water, 4% carbohydrate (CHO), 6% CHO, or 8% CHO. Gastric volume was determined prior to each drink and at 90 min using the modified double-sampling technique. Gross gastric volumes were significantly greater and mean gastric emptying rates and the percentage of ingested beverage emptied from the stomach were significantly less for 8% CHO. These data indicate that repeated ingestion of an 8% CHO beverage during exercise significantly reduces gastric emptying rate, whereas lower concentrations of carbohydrate do not. In addition, beverage osmolality is not as important as beverage energy content in influencing gastric emptying rate at these carbohydrate concentrations. PMID- 10477363 TI - Calcium losses resulting from an acute bout of moderate-intensity exercise. AB - The purpose of this study was to compare daily calcium (Ca) losses in sweat (S) and urine (U) on an exercise day (E) with losses on the preceding day (i.e., a rest day) during which no exercise (NE) was performed. Ten healthy male volunteers (23.9 +/- 3.2 years) performed a single bout of moderate exercise (running at 80% HRmax) for 45 min in a warm (32 degrees C, 58% relative humidity) environment on E. When E and NE were compared, neither Ca intake (1,232 +/- 714 and 1, 148 +/- 482 mg, respectively) nor urinary Ca excretion (206 +/- 128 and 189 +/- 130 mg, respectively) were different (p >.05). Sweat Ca losses during the exercise bout averaged 45 +/- 12 mg. The results indicate that, although a small amount of Ca is lost in sweat during 45 min of moderate-intensity exercise, measured (sweat and urine losses combined) Ca losses (251 +/- 128 and 189 +/- 130 mg) were not different (p >.05) between days (E and NE, respectively). These data suggest that moderate exercise for up to 45 min in a warm, humid environment does not markedly increase Ca intake requirements. PMID- 10477365 TI - Surgery with computerized virtual reality for the automatic detection of tumors. AB - We present a novel and highly accurate system based on informatics engineering capable of automatic detection of tumors directly in the operating field. The system can identify the outlines of the tumor, determine whether it is malignant or not, detect lymphadenopathy and determine whether nodes are metastasized or not. The highly elaborate system, based on artificial vision, has been used in 30 gastric and 5 pancreatic neoplasms, among other tumor types. Images of the surgical field were recorded with a video camera connected to a computer, which was operated by the engineer. Questions asked by the surgeon during the procedure were processed immediately and sent to the virtual reality helmet worn by the surgeon, to the TV monitor in the operating room, or to both. The system is based on purely physical and mathematical processes that work reliably; in this sense it is free from errors and is self-consistent, operator errors or hardware failure excepted. In all cases tested here the system correctly identified the tumor as benign or malignant, revealed the extension of the tumor, and detected lymph node metastases. In every case these results were confirmed by histological examination. PMID- 10477364 TI - Decreased resting metabolic rate in ballet dancers with menstrual irregularity. AB - We studied 21 ballet dancers aged 19.4 +/- 1.4 years, hypothesizing that undernutrition was a major factor in menstrual irregularity in this population. Menstrual history was determined by questionnaire. Eight dancers had always been regular (R). Thirteen subjects had a history of menstrual irregularity (HI). Of these, 2 were currently regularly menstruating, 3 had short cycles, 6 were oligomenorrheic, and 2 were amenorrheic. Subjects completed a weighed dietary record and an Eating Attitudes Test (EAT). The following physiological parameters were measured: body composition by anthropometry, resting metabolic rate (RMR) by open-circuit indirect calorimetry, and serum thyroid hormone concentrations by radioimmunoassay. R subjects had significantly higher RMR than HI subjects. Also, HI subjects had lower RMR than predicted by fat-free mass, compared to the R subjects. Neither reported energy intake nor serum thyroid hormone concentrations were different between R and HI subjects. EAT scores varied and were not different between groups. We concluded that in ballet dancers, low RMR is more strongly associated with menstrual irregularity than is current reported energy intake or serum thyroid hormone concentrations. PMID- 10477366 TI - Colon cancer: p53 expression and DNA ploidy. Their relation to proximal or distal tumor site. AB - Overexpression of nuclear p53 and DNA ploidy were analyzed in a series of 65 colorectal adenocarcinomas and correlated with standard clinical and pathological variables (Dukes stage, tumor site, histological grade and type, and nature of the tumor margins). Immunohistochemical tests were done with the DO-7 monoclonal antibody, using formalin-fixed tissue samples and an antigen retrieval solution. Levels of p53 expression were evaluated using a semiquantitative grading system (CAS 200, BD). Nuclear staining of more than 15% of neoplastic cells was observed in 35 samples (53.8%), which were classified as p53-positive. DNA content was measured by flow cytometry in samples of fresh tissue. Tumor site had a significant direct relationship with DNA ploidy (p < 0.01) and p53 expression (p < 0.001). Proximal tumors were more frequently diploid than were distal tumors (78.6% vs 32%). Moreover, distal neoplasms showed more p53 expression than proximal tumors (64.6% vs 14.3%). However, there was no correlation between the other clinical or pathological variables and the pathological parameter p53 expression and DNA ploidy. Our data support the hypothesis that mechanisms of colorectal carcinogenesis may differ in proximal and distal neoplasms. PMID- 10477367 TI - Cleaning and disinfection of gastrointestinal endoscopes. Comparative analysis of two disinfectants. AB - Gastrointestinal endoscopy can lead to infectious complications, and endoscopes must be disinfected to prevent them. AIM: to evaluate three methods of disinfection: 1) usual cleaning technique and immersion in glutaraldehyde phenolate (GP); 2) meticulous cleaning and immersion in GP, and 3) meticulous cleaning and immersion in hydrogen peroxide. METHOD: thirty endoscopes (15 gastroscopes and 15 colonoscopes) were disinfected with each method. Samples were taken following endoscopic exploration, after cleaning and after disinfection, and were cultured. The number of positive culture (cfu/ml > 1) was counted. RESULTS: the rate of contamination of endoscopes did not decrease significantly after cleaning with method 1 (66 vs 60%), but did decrease with method 2 (38 vs 16%) and method 3 (53 vs 17%). The contamination rate after cleaning was significantly lower with methods 2 and 3 (p < 0.005). This rate was also lower after disinfection (p < 0.025). Method 3 achieved 0% contamination following disinfection. CONCLUSIONS: the greatest decrease in contamination rate was achieved with conscientious cleaning followed by disinfection. Both disinfectants yielded similar results, although hydrogen peroxide produced a higher level of disinfection. PMID- 10477368 TI - Influence of nonsteroidal anti-inflammatory drugs on clinical course in upper gastrointestinal tract bleeding. AB - To establish the influence of nonsteroidal anti-inflammatory drugs (NSAIDs) on the clinical course of upper gastrointestinal bleeding (UGIB), we designed a longitudinal cohort study to assess the effect of prior use of these drugs on several outcome variables in patients with UGIB. RESULTS: 164 (46.6%) of 352 patients with UGIB consumed NSAIDs in the week previous to the onset of bleeding. This group had significantly fewer previous episodes of peptic ulcer and UGIB (p < 0.01), more associated comorbidity, and increased use of steroids (p < 0.05). On endoscopic examination, gastric ulcer was the most frequent lesion in patients with NSAIDs as compared to duodenal ulcer in patients without NSAID use (p < 0.0001). There were no differences in the multivariate analysis between groups regarding lowest hemoglobin level, need for transfusion, stability of bleeding, surgical or endoscopic management, depth of lesions and hospital stay. CONCLUSIONS: a relevant proportion of patients admitted for UGIB had used NSAIDs during the previous week. The most frequent lesion in this group was gastric ulcer. In our study the clinical course of NSAID-associated UGIB was similar to that in other nonvariceal types of bleeding. PMID- 10477369 TI - [Are all H. pylori "bad"?]. AB - The infection by Helicobacter pylori arises a question of great interest: should the infection be considered a disease in all cases? or, on the contrary, is the microorganism harmful in only some circumstances? The response to these questions is followed by therapeutic implications of great importance, as it is followed by the decision of administering eradication therapy to every patient or only to some individuals. In this article the arguments supporting each of the aforementioned options are reviewed. Firstly, the aphorism recently stated <> suggests that all microorganisms are harmful and, therefore, generalized H. pylori eradication therapy should be administered to all infected subjects. Among the arguments supporting this attitude are the following: the H. pylori <> carrier status does not exist; a high risk of developing peptic ulcer exists in infected patients; H. pylori positive patients are at risk of suffering gastric adenocarcinoma and lymphoma; and, finally, the eradication of the microorganism would reduce the potential reservoir of the infection. On the other hand, the expression <> summarises a more conservative position, that suggests the existence of <>, <> H. pylori, advising the eradication of the microorganism in only some cases. Several arguments supporting this idea, and, therefore, against the generalized use of eradication therapy, have been proposed: H. pylori has colonized humans since immemorial time and, therefore, it can not be very harmful to the human; the bacterium will induce peptic ulcer or gastric cancer in only a minority of infected subjects; the beneficial effect of eradication has been definitively demonstrated in few entities as gastroduodenal ulcer and MALT lymphoma; some strains could be beneficial to the human; generalized administration of antibiotic therapy will cause problems; the H. pylori vaccine represents a more rational alternative; and, finally, the incidence of H. pylori infection is spontaneously decreasing in some regions. In summary, there are strong arguments supporting both of the strategies that are reviewed in the present article, and, therefore, the dilemma <> remains. PMID- 10477370 TI - [Sump syndrome. Treatment by endoscopic biliary "rendez-vous" procedure]. AB - Sump syndrome consists on an entity in which the distal portion of the biliary tree does not drainage by the major papilla, so food and bile are retained and infections could appear leading to cholangitis. The real incidence of the sump syndrome after side-to-side choledocoduodenostomy is unknown, but has been described less than 10%. Several procedures have been used to solve it but at present endoscopic sphincterotomy is of choice the treatment of this condition that is followed by a good outcome. When major papilla can not be negotiated, an endoscopic approach consisting in a biliary endoscopic <> procedure could be used, negotiating the biliary tree by the side-to-side choledocoduodenostomy in this case. We report a clinical case of a patient with a side-to-side choledocoduodenostomy because of choledocolithiasis who developed a sump syndome. He was treated by a biliary endoscopic <> procedure because a papillary stenosis did not allow a transpapillar access to the biliary tree. PMID- 10477371 TI - A peculiar increase in frequency of medical emergencies after opening of a war isolated enclave in Bosnia. PMID- 10477372 TI - [Low-grade MALT gastric lymphomas: antibiotics or gastrectomy]. PMID- 10477373 TI - [Acute acalculous cholecystitis following aortic surgery]. PMID- 10477374 TI - [Vesico-appendicular fistula caused by appendicitis]. PMID- 10477375 TI - [The word pouchitis]. PMID- 10477376 TI - [Dietary fibre intake and colon cancer]. AB - This paper summarizes the contradictory epidemiological data concerning the association between vegetal fibres intake and colorectal cancer incidence. The key to these discrepancies might reside in the different amounts and ratios of butyrate and other short chain fatty acids (SCFA) produced by colonic anaerobic bacteria during fermentation of fibers from different origins, wheat bran fibers being better producers and therefore more protective. Butyrate, an established histone deacetylase inhibitor, acts by favoring an active chromatin configuration of the P21WAF1/CIP1 gene which, as an inhibitor of cyclin-dependent protein kinases, blocks the cell cycle. PMID- 10477377 TI - [PKC 412]. PMID- 10477378 TI - [Androgen-independent prostate carcinoma and androgen-receptor: recent progress in molecular genetics]. AB - Prostate cancer is an androgen-dependent tumor which presents an androgen independent regrowth after clinical regression in response to antiandrogen treatment. Four hypotheses have been developed to understand how androgen signal transduction pathway mediate androgen-independent tumor progression: over expression of the wild-type androgen-receptor gene, androgen-receptor gene mutation, excessive recruitment of transcriptional co-activator ARA-70 and a cross-talk between the androgen-receptor and the growth factor receptor pathways. In this work, C. Sawyers's group elegantly demonstrates, in LAPC-4 androgen independent prostate cancer sublines, that forced hyperexpression of HER-2/Neu receptor tyrosine kinase allowed androgen-independent growth, that HER-2/Neu activated the androgen-receptor pathway in the absence of androgens and synergized with low levels of androgen to superactivate the pathway. These important data could have therapeutic implications for the management of androgen independent prostate cancer. PMID- 10477379 TI - [Surrogate endpoints]. AB - The main endpoint of a phase III trial in oncology must be relevant clinically. In practice, this endpoint is either the disease-free or the overall survival, which requires both long term follow-up and a large number of patients. In phase II trials, it is essential to proceed rapidly, and one therefore uses a surrogate endpoint. Surrogate endpoints are adequate for phase II trials. Their use for phase III trials should be restricted to the rare situations where their validity has been established for the therapeutic and a under study. This surrogate endpoint must be faster to obtain than the clinical endpoint it replaces. Ideally, it should capture all of the treatment effect on the main endpoint, i.e. there should be no effect of the treatment on survival, once the value of the surrogate endpoint is known. PMID- 10477380 TI - [Merkel cell carcinoma: diagnostic and treatment]. AB - Merkel cell carcinoma (MCC) is an uncommon neuro-endocrine tumor of the sun exposed skin predominantly observed in white patients in the sixth decade of the life. In electron microscopy MCC characteristically contains dense core secretory granules. This tumor expresses both epithelial (keratins of low molecular weight of the simple epithelial type, epithelial membrane antigen) and neuro-endocrine markers (neuron-specific enolase, chromogranin A). The analysis of available literature indicates that MCC has a propensity for involvement of regional lymphatics (12% of patients at the date of initial diagnosis and 50 to 66% afterwards) and for metastases to distant sites (20 to 52%). The prognosis of metastatic disease is poor. The overall survival from the diagnosis of metastases is 6 months. The optimal treatment of locoregional stages should include excision and post-operative radiation. The role of chemotherapy in this tumor is unclear but recent data suggest a high chemosensitivity. PMID- 10477381 TI - [Updating 1999 of Standards, Options and Recommendations (SOR) for the clinical use of erythropoietin in oncology. FEDERATION OF THE FRENCH CANCER CENTRES (FNCLCC)]. AB - CONTEXT: The <> (SOR), started in 1993, are a collaborative project between the Federation of the French Cancer Centres (FNCLCC), the 20 French Cancer Centres and specialists from French Public Universities, General Hospitals and Private Clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and outcomes for cancer patients. The methodology is based on literature review and critical appraisal by a multidisciplinary experts group, with feedback from specialists in cancer care delivery. The initial guidelines are being updated in case of new evidence. OBJECTIVES: To update the clinical practice guideline [39] with definitions of new Standards, Options and Recommendations for the use of recombinant human erythropoietin (rHuEPO) in oncology. METHODS: Data have been identified by literature search using Medline, Current Contents, Embase, Cancerlit (march 1996-march 1999). The main end points considered were hemoglobin level, haematocrit, quality of life, transfusion requirements, incidence and length of hospital stays, efficacy of cancer treatment, safety and costs. Once the guideline was updated and defined, the document was submitted to 42 reviewers for peer review, and to the medical committees of the 20 French Cancer Centres for review and agreement. RESULTS: The new key recommendations are: 1) The use of recombinant human erythropoietin in oncology is an alternative to treat chemotherapy-induced anemia when the chemotherapeutic regimen contains platinum; 2) Cancer-induced anemia reduces patients' quality of life. Treatment of anemia by transfusions of erythropoietin may improve quality of life; 3) We recommend assessment of haemoglobin levels during radiation therapy and the possible use of erythropoietin to optimise the efficacy of radiation therapy; 4) Erythropoietin is effective in others pathologies (multiple myeloma, non-Hodgkin lymphoma, non platinum based chemotherapy.) and also in pediatric patients but the risk/benefit ratio for anemia therapy (i.e. transfusion or erythropoietin therapy) must be analysed for each individual; 5) We recommend an economic analysis of the need of erythropoietin within the context of the french health care system. PMID- 10477382 TI - [Standards, Options and Recommendations (SOR) for good practices in dentistry for head and neck cancer patients. Federation of the French Cancer Centres (FNCLCC)]. AB - CONTEXT: The Standards, Options and Recommendations (SOR), initiated in 1993, is a collaborative project between the Federation of the French Cancer Centres (FNCLCC), the 20 French Cancer Centres and specialists from French Public Universities, General Hospitals and Private Clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and outcomes for cancer patients. The methodology is based on literature review and critical appraisal by a multidisciplinary experts group, with feedback from specialists in cancer care delivery. OBJECTIVES: To develop clinical practice guidelines for dentistry and oral hygiene in head and neck cancer patients. METHODS: Data have been identified by literature search using Medline (up to January 1999) and personal reference lists. The main end points considered were risk factors for treatment related late effects, safety and quality of life, efficacy of dental preventative measures and treatment. Once the guidelines were defined, the document was submitted to reviewers for peer review and to the medical committees of the 20 French Cancer Centres for review and agreement. RESULTS: The key recommendations are: 1) before receiving radiotherapy, surgery and chemotherapy for head and neck cancer, patients must benefit from a multidisciplinary approach including dental evaluation; 2) the patients must be informed of precautions and educated about oral hygiene; 3) after radiotherapy, the most important dental late effect to prevent is radionecrosis, in accordance with the oral and dental state, the dentist may propose conservation or extraction of teeth, fluoridation and regular follow-up; 4) during chemotherapy, the principal complications are mucositis, haemorrhage and infection risk; 5) after surgery, the dentist may propose prosthetic measures with the aim functional, aesthetic and psychological benefit; 6) in the particular case of children, treatment and prevention are the same as for adults but the follow-up is specific because of the dental development. PMID- 10477383 TI - [First treatment for brain metastases by stereotactic radiosurgery]. AB - Local control of brain metastases is better with first treatment by stereotactic radiosurgery than with radiosurgery for recurrence. We reported a retrospective analysis of the influence of clinical and technical factors on local control and survival after radiosurgery realised in first intention. From January 1994 to December 1997, 26 patients presenting with 43 metastases underwent radiosurgery. The median age was 61 years and the median Karnofsky index 70. Primary sites included: lung (12 patients), kidney (7 patients), breast (2 patients), colon (1 patient), melanoma (2 patients), osteosarcoma (1 patient), it was unknown for one patient. Seven patients had extracranial metastases. Twenty-one sessions of radiosurgery have been realized for one metastase, and 9 for two, three or four lesions. The median diameter was 21 mm and the median volume 1.8 cm3. The median peripheral dose to the lesion was 14 Gy, and the median dose at the isocenter 20 Gy. Forty-two metastases were evaluable for response analysis. The overall local control rate was 90.5% and the 1-year, 2- and 3-year actuarial rates were 85% and 75%. In univariate analysis, theorical radioresistance was significantly associated with better local control (100% versus 77%, p < 0.05). All patients were evaluable for survival. The median survival rate was 15 months. Four patients had a symptomatic oedema (RTOG grade II). Two lesions have required a surgical excision. In conclusion, low dose radiosurgery (14 Gy delivered at the periphery of metastasis) can be proposed in first intention for brain metastases, in particularly for theorical radioresistant lesions. PMID- 10477384 TI - [Intraperitoneal high dose chemotherapy as consolidation treatment for advanced ovarian carcinoma: a pilot study]. AB - Consolidation treatment of advanced ovarian carcinoma, especially the place of intraperitoneal chemotherapy, remains a controversial subject. From January 1988 to July 1995, 39 patients, median age 54 years, received intraperitoneal chemotherapy as consolidation treatment after second-look surgery. At the time of intraperitoneal chemotherapy, 30 patients had no residual disease. Intraperitoneal drug administration used a Tenckoff catheter or a lumbar needle. Treatment combined 5 fluorouracil 1 g/m2 and cisplatin 200 mg/m2, associated with a systemic sodium thiosulfate rescue as nephroprotector. A pharmacological analysis was done for 9 patients: the exposure of peritoneal cavity to cisplatin exceeded that of the plasma by 11 fold. Hematologic and nephrologic toxicity were acceptable. The median follow-up is 43 months. The disease free survival is 36,6 months, but 48,5 months if no residual disease at the time of intraperitoneal chemotherapy. Consolidation treatment by intense intraperitoneal chemotherapy is a feasible approach and might be beneficial in chemosensitive patients devoid of macroscopic remnants, but must be compared with others approaches. PMID- 10477385 TI - [Standardization and quality control in the evaluation of proliferation parameters in T1T2, N0N1, M0 breast cancer : multicentric retrospective study I. DNA synthesis enzyme activities]. AB - As part of a clinical research project co-ordinated in Grenoble, six French institutions (CRLCC Angers, CHU Grenoble, Hospices civils Lyon, AP Marseille, CRLCC St-Cloud, CHU Tours) grouped together in order to study the following proliferative parameters in primary breast cancer: DNA synthesis enzymes [thymidine kinase (TK), thymidylate synthase (TS)], signal transduction enzyme [protein tyrosine kinase (PTK)] and S-phase fraction (%S). TK, TS and PTK were measured in cytosols using radio-enzymatic biochemical methods. S-phase was estimated using flow cytometry. The first step consisted in standardization and technical validation of the measurements. The second step consisted in the clinical validation by using a retrospective series of 1,003 breast cancers T1T2, N0N1, M0. We report the results of the first step, together with the distributions of the variables and their relationship with classical clinical variables: 1) Using standardized methods and a cytosolic control, a good reproducibility of measurements was obtained, whether assays were performed in one (TS, PTK) or in several laboratories (TK). 2) Significantly different distributions of TK and TS were observed between the different centres mainly due to different conditions of storage of tumours and cytosols. 3) A highly significant correlation was observed between TK, TS and PTK. Highest TK, TS and PTK levels were observed in tumours with high histological grade or receptor negative tumors. This study clearly illustrates the importance of quality assurance of multicentre studies. PMID- 10477386 TI - [Standardization and quality control in the evaluation of proliferation parameters in T1T2, N0N1, M0 breast cancer: multicentric retrospective study II. DNA-ploidy and S-phase fraction]. AB - As part of a clinical research project, proliferative parameters were studied in primary breast cancer: standardization and technical validation of thymidine kinase (TK), thymidylate synthase (TS) and protein tyrosine kinase (PTK) are described. A total of 633 frozen tumor specimens, available in four institutions, was analyzed in three flow cytometry laboratories for DNA content and percentage of S-phase cells (%S) measurement. 1) The standardization step consisted in developing a common protocol for sample preparation; then, common cell suspensions were analyzed in order to perform an inter-laboratory control. Objective guidelines were elaborated to interpret DNA histograms in breast carcinoma. 2) DNA-aneuploidy was observed in 61% of cases of the retrospective series. Compared with DNA-aneuploid tumors, mean %S was significantly lower in case of DNA-diploidy (respectively: 6.4% and 2.2%, p < 0.001). When compared between the four institutions, %S distributions did not differ significantly. 3) %S is strongly correlated with TK, TS and PTK and high percentages were also observed in high grade tumors or tumor without hormone receptors. These results show that a standardization in using flow cytometers and DNA software allows multicenter studies. PMID- 10477387 TI - [Spring American Congress 1999. 1. AACR 90th annual congress (Philadelphia, 10-14 April, 1999). American Association of Cancer Research]. PMID- 10477388 TI - Adenosine - cyclic AMP pathways and cytokine expression. AB - Adenosine and cAMP are potent modulators of immune-triggered cytokine production. Their effects overlap with regard to the inhibition of the pro-inflammatory cytokines TNF-alpha, IFN-gamma, IL-12, and the stimulation of production of the major anti-inflammatory cytokine IL-10. They may tentatively be considered to be upregulators of the production of Th2 cytokines (IL-10, IL-6), but downregulators of the production of Th1 cytokines (IL-2 and IFN-gamma). Cytokines produced in common by Th0, Th1 and Th2 cells are affected as well, although the low quantity and heterogeneity of the contemporary experimental data do not allow unambiguous conclusions to be drawn. Nevertheless, IL-3, IL-4, MIP-1alpha/beta and GM-CSF have usually been found to be inhibited, IL-5 stimulated, while IL-1 remains largely unaffected by adenosine or cAMP. These effects, and in particular the inhibition of TNF-alpha and stimulation of IL-10 expression, might be of therapeutic value in a variety of pathophysiological conditions. PMID- 10477389 TI - Induction and large-scale expansion of CD8+ tumor specific cytotoxic T lymphocytes from peripheral blood lymphocytes by in vitro stimulation with CD80 transfected autologous melanoma cells. AB - Human melanoma cell lines may induce a specific T cell response against tumor cells in vitro. However, after repeated restimulation with autologous tumor cells, expansion of CTL is limited and often apoptosis of the T cells occurs. In order to improve conditions inducing primary T cell responses and thus allowing further expansion of tumor specific T cells for an adoptive transfer, we transfected human melanoma cells with the B7.1 gene (CD80), known to be a potent costimulatory molecule for T cell activation. CD80 expression on melanoma cells resulted in improved primary T cell activation, especially of CD8+ T cells. Furthermore, restimulation with CD80+ tumor cells gave rise to long term proliferating CD8+ T cell lines demonstrating an 100-fold expansion of T cells compared to the 20-30-fold increased numbers obtained with the controls (parental tumor cells +/- anti-CD28). T cells stimulated with CD80+ melanoma cells were found to display a MHC class I-restricted cytotoxic activity against the autologous tumor cells. In conclusion, these studies demonstrate the requirement of costimulation in generating large numbers of tumor specific T cells in vitro that may be used for an adoptive transfer in tumor immunotherapy. PMID- 10477390 TI - Interleukin-6 receptor signaling. II. Bio-availability of interleukin-6 in serum. AB - Interleukin-6 (IL-6) is used as a growth factor by various tumor cells. It binds to a gp80 specific receptor (IL-6R) and then to a gp130 transducing chain. Both receptor chains are released as soluble functional proteins which circulate in biological fluids. With a view to studying the physiological role of these soluble receptors, both proteins were purified from human plasma. Surface plasmon resonance was used to measure the kinetic constants of equilibria between IL-6 and natural sIL-6R, and between the IL-6/sIL-6R complex and soluble gp130. Kd values were found to be 0. 9 and 2.3 nM respectively. Soluble natural IL-6R and gp130 were also found to interact with a Kd of 2.8 nM in the absence of IL-6. By using these Kd values, a mathematical simulation predicted that 1) within a large range of IL-6, sIL-6R and sgp130 concentrations, free IL-6 represents 30% of the total circulating cytokine, 2) sIL-6R overconcentrations lead to dramatic changes of the concentration of free IL-6, 3) increased concentrations of sgp130 should produce an efficient buffering effect on the IL-6/sIL-6R complex without incidence on the level of free IL-6. According to this model, the IL-6/sIL-6R complex appears to be an important support of IL-6 signaling in the most commonly encountered in vivo situations. The concentration of this complex is directly under the control of the concentration of sIL-6R; its bio-availability should be efficiently buffered by increased sgp130 concentrations. PMID- 10477391 TI - Subtherapeutic doses of interleukin-15 augment the antitumor effect of interleukin-12 in a B16F10 melanoma model in mice. AB - Interleukin-12 (IL-12) is a potent immunoregulatory cytokine that exhibits antitumor activity in many experimental tumor models. In the present study, we investigated the ability of IL-15, a cytokine sharing many functions of IL-2, to modulate antitumor effectiveness of IL-12 against B16F10 melanoma in mice. In a model of locally growing tumor, intratumoral (i.t.) administration of IL-12, in three cycles of five consecutive daily injections (0.1 mug) followed by 2 days of rest, led to considerable delay of tumor development but no curative response was achieved. When combined with IL-12, subtherapeutic doses of IL-15 (0.4 mug) pontentiated the antitumor effects of IL-12 and induced complete tumor regressions in 50% of mice. Similar results were obtained in a model in which tumor-bearing mice were intravenously co-injected with melanoma cells to induce metastases. Combined administration of IL-12 and IL-15 yielded greater antitumor activity than injections of either cytokine alone and resulted in prolonged survival of mice bearing locally growing tumor and metastases. Studies of immunological parameters in mice treated with both IL-12 and IL-15 have shown enhanced NK activity (against YAC-1 cells) in the spleen and stimulation of both NK activity and specific anti-B16F10 cytotoxic effector cells in tumor-draining lymph nodes (LN). The strong antitumor effect of the IL-12 + IL-15 combination correlated with a high serum level of IFN-gamma in the treated mice. Moreover, increased expression of IL-15Ralpha was demonstrated in LN lymphocytes isolated from mice injected with IL-12. This result together with findings of other authors showing enhanced expression of IL-12 receptor by IL-15 [1] suggests that the augmentation of the antitumor effect during the course of IL-12/IL-15-based therapy could result from reciprocal upregulation of receptors by both cytokines and synergistic effects on IFN-gamma induction. PMID- 10477392 TI - Effect of tumor growth factor-beta on NK receptor expression by allostimulated CD8+ T lymphocytes. AB - We have examined the influence of the immunosuppressive cytokine TGF-beta on NK receptor expression by T lymphocytes upon allogeneic activation. Using the primary mixed lymphocyte reaction (MLR), our data show that allostimulation induced the expression of CD94/NKG2-A on alloactivated CD8+ T cells. This expression was increased in the presence of TGF-beta whereas IL-15 had no significant effect. The blockage of CD94 and NKG2-A resulted in increased lysis of targets by alloactivated cytotoxic T cells. This increase was dependent on the activation state of T cells. Using PCR, we also demonstrated that TGF-beta had no effect on the transcription of non-inhibitory NKG2 molecules. The present results show that allostimulation can induce CD94 and further point out the role of TGF beta in the induction of the CD94/NKG2-A receptor on alloactivated T cells. PMID- 10477393 TI - The intestinal microflora regulates cytokine production positively in spleen derived macrophages but negatively in bone marrow-derived macrophages. AB - Besides its role as a barrier against potential pathogens, intestinal flora is presumed to protect the host by priming the immunological defense mechanisms. In this respect, the influence of intestinal flora on macrophage precursors was examined, and its modulating effect was compared on LPS-induced cytokine production by macrophages derived from bone marrow and spleen precursors (BMDM and SDM respectively). The regulation of IL-1, IL-6, TNF-alpha and IL-12 production in macrophages from germ-free and from three groups of flora associated mice, conventional, conventionalized and E. coli-mono-associated mice, was investigated. The whole flora inhibited IL-1, TNF-alpha and IL-12 secretion by BMDM, whereas it had a stimulatory effect on IL-12 secretion by SDM. Implantation of E. coli alone enhanced cytokine secretion by BMDM but had a more limited effect than whole flora on SDM, enhancing only TNF-alpha and IL-12 secretion. Study of expression of mRNA showed a correlation with protein secretion for IL-6 but not for TNF-alpha and IL-1. IL-12 enhancement in BMDM seemed to be dependent on regulation of p35 mRNA expression while it was correlated to increased p40 mRNA expression in SDM. The results demonstrated that intestinal flora modulated bone marrow and spleen macrophage cytokine production in a differential manner and suggested a role for bacteria other than E. coli among the whole flora. The contrasting effects exerted by the intestinal flora on bone marrow and spleen precursors are an interesting observation in view of the different functions of these organs in immunity. The finding that intestinal flora enhanced IL-12 production in spleen is also potentially important since this cytokine is implicated in the determination of the relative levels of Th1 and Th2 responses and plays a pivotal role in host defense against intracellular microorganisms. PMID- 10477394 TI - Impairment of HIV polymorphonuclear leukocyte transmigration across T84 cell monolayers: an alternative mechanisms for increased intestinal bacterial infections in AIDS? AB - Our objective was to study the influence of HIV infection of polymorphonuclear leukocytes (PMN) on transepithelial migration. To date, reports of functional PMN chemotaxis in AIDS are contradictory. This is the first attempt to assess this function via an in vitro model allowing transmigration of neutrophils through an intestinal epithelial barrier. PMN were isolated from 45 HIV-infected patients and 45 healthy volunteers. PMN transmigration across T84 epithelial cells was initiated by applying either various concentrations of formyl-met-leu-phe peptide (f-MLP) or interleukin-8 and assayed by quantification of myeloperoxidase activity. CD11b, CD18, and CD47 expression on PMN was compared before and after transepithelial migration by flow cytometry analysis. CD11b expression was studied by electron microscopy. Apoptosis of transmigrated HIV PMN and control PMN was investigated by morphology and DNA fragmentation characterization. Compared to control PMN, HIV PMN exhibited a decrease in transepithelial migration that directly correlated with CD4+ counts. Basal and transepithelial migration-mediated expression of CD11b, CD18, and CD47 were unmodified in HIV PMN compared to control PMN. Electron microscopy labeling confirmed no difference in CD11b expression on HIV and control PMN. The index of apoptosis in transmigrated HIV PMN and control PMN was identical. These data provide evidence of a defect in the f-MLP-induced chemotaxis of PMN from HIV-infected patients across an intestinal epithelial barrier. This defective migration is not due to a quantitative modification of CD11b, CD18 and CD47 on HIV PMN suggesting a more subtle alteration. The impairment in the transmigration function may contribute in vivo to an increased susceptibility to intestinal bacterial infection in HIV infected patients. PMID- 10477395 TI - Rat tumour necrosis factor-alpha: expression in recombinant Pichia pastoris, purification, characterization and development of a novel ELISA. AB - Tumour necrosis factor-alpha is a pro-inflammatory cytokine involved in many aspects of acute phase and immune responses. Species specificity in the biological action and receptor binding of TNF-alpha make it desirable to use homologous reagents in experimental models, both in vivo and in vitro. As the rat is the model of choice in many investigations on fever, trauma and pathology, there is a need for specific rat reagents. In this paper, we describe the production of recombinant rat TNF-alpha in milligram quantities, using a methylotrophic yeast expression system, Pichia pastoris. Recombinant TNF-alpha was produced intracellularly in a soluble form, cells were lysed and the protein purified by ammonium sulphate precipitation, Sephadex G75 fractionation and finally, ion-exchange chromatography. The purified recombinant rat TNF-alpha had a molecular mass of 17401.38 +/- 0.38 Da, which is within 1 Da of the value predicted by the sequence data, taking into account N-acetylation of the initial methionine residue and a single disulphide bridge between amino acids 70 and 101. Recombinant rat TNF-alpha was shown to be 20 x fold more biologically active in the WEHI cytotoxicity assay, than the human standard preparation. Polyclonal antibodies were raised against purified recombinant rat TNF-alpha, these reagents were used to develop a novel enzyme-linked immunosorbant assay (ELISA). The ELISA was sensitive to 10 pg.ml- 1 rat TNF-alpha and was specific for TNF-alpha, showing no cross-reactivity with rat IL-1alpha, rat IL-1beta, rat IL-1Ra or rat IL-6. The ELISA was used to measure TNF-alpha in the plasma of rats injected with bacterial endotoxin and in cultures of rat white blood cells. The ELISA was shown to be a robust method suitable for use in assaying samples generated in both in vivo or in vitro experiments. PMID- 10477396 TI - Cellular distribution and relative amounts of vascular endothelium growth factor mRNA in granulosa cells from human preovulatory follicles. AB - Cells were obtained from patients undergoing in vitro fertilization. They were cultured and those producing vascular endothelium growth factor (VEGF) were detected by flow cytometry; relative amounts of mRNA were detected by RT-PCR and measured by PCR Elisa after RT-PCR products were biotinylated. Most of the granulosa cells produced VEGF. This production was maintained over 5 days in culture without adding hCG. The two diffusible forms, VEGF 121 and 165, were the most abundant. VEGF 145, which is specific to the reproductive system, was less abundant. VEGF 189, which is not freely secreted, was not produced by granulosa cells; small amounts were only detected in preparations containing leukocytes. TNF-alpha decreased VEGF production; the effect of TNF-alpha was neutralized by 10 nM staurosporine. Thus, the VEGF in human preovulatory follicles is mostly in the granulosa cells. These cells are therefore a major source of VEGF at ovulation and may play a key role in physiological and pathological processes which involve changes in vascular permeability and/or angiogenesis. The data also suggest that TNF-alpha via protein kinase C modulates the production of VEGF. PMID- 10477397 TI - The role of interferon regulatory factor-1 in cytokine-induced mRNA expression and cell death in murine pancreatic beta-cells. AB - Combinations of cytokines, including interleukin-1beta (IL-1beta) and interferon gamma (IFN-gamma), induce nitric oxide (NO) production and cell death in pancreatic islet cells. We have previously shown that these events are preceded by increased expression of the transcription factor interferon regulatory factor 1 (IRF-1). We utilized an IRF-1 knockout mouse (IRF-1-/-) to investigate the role of IRF-1 in cytokine-induced islet- and beta-cell gene expression and cell death. For this purpose, pancreatic islets or FACS-purified beta-cells were isolated from wild type (wt) or IRF-1-/- mice. These cells were exposed for different time points to IL-1beta (50 U/mI), IFN-gamma (1,000 U/ml) and/or TNF-alpha (1,000 U/ml) before being harvested for determination of viability (by nuclear dyes) and mRNA expression (by RT-PCR with specific primers). Following a 24 hours exposure to IL-1beta or IL-1beta + IFN-gamma, pancreatic islets isolated from IRF-1-/- mice presented a 30-50% reduction in medium nitrite accumulation and inducible NO synthase (iNOS) expression. Interestingly, both wt and IRF-1-/- purified beta cells failed to produce NO in response to IL-1beta alone, but presented a similar increase in nitrite accumulation and iNOS expression following exposure to IL 1beta + IFN-gamma. The basal expression of MHC class I mRNA was lower in IRF-1-/- islet cells (30% reduction), but there was a similar 2-4 fold-increase in MHC expression in islet cells from both strains following cytokine exposure. IL-1beta induced serine protease inhibitor-3 (SPI-3; a putative cellular "defense" protein) mRNA expression in both wt and IRF-1-/- islets or beta-cells. IFN-gamma decreased the IL-1beta-induced SPI-3 expression in wt islets or beta-cells, but induced a 5-fold increase in the expression of this mRNA in IRF-1-/- islets cells, suggesting that IRF-1 mediates an inhibitory effect of IFN-gamma on SPI-3 expression. Treatment of whole islets for 3 days with IL-1beta + IFN-gamma induced significantly more islet cell death in wt than in IRF-1-/- mice (respectively 85 +/- 3% versus 31 +/- 4% dead cells). On the other hand, prolonged exposure (3-9 days) of FACS-purified beta-cells to the same cytokines, or a combination of 3 cytokines, led to a similar increase in cell death in both IRF-1-/- and wt islets. In conclusion, IRF-1 contributes to cytokine-induced islet iNOS expression and cell death. These effects are absent in purified beta cells, suggesting that IRF-1 may mediate its effects on whole islets via activation of non-endocrine cells (e.g. macrophages and ductal cells) present in these preparations. PMID- 10477398 TI - Development of antibodies to interferon beta in patients: technical and biological aspects. AB - There are now several papers describing the development of antibodies to interferons (IFN) in patients undergoing IFN therapy. Moreover, there is increasing evidence to indicate that the development of antibodies to IFN may be associated with a failure of the beneficial effects of the therapy. This paper will review and discuss what is currently known about the technical, and biological aspects of antibodies to IFN, with particular reference to antibodies to IFN beta that develop during therapy. Three main considerations arise from the data. Firstly, a standardized quantitative assay to detect antibody to IFN must be agreed upon. Only when results can be compared, both qualitatively and quantitatively, will it be possible to monitor fully the ability of antibodies to cause a relapse during treatment. Secondly, sufficient data are now available to provide a rationale for monitoring the presence of anti-IFN antibodies in patients treated with IFN. This approach may allow a better understanding of the disease reactivation state observed in numerous patients treated with IFN. Finally, approaches aimed at limiting the immunogenicity of IFN preparations and/or strategies designed to circumvent antibody-mediated resistance to IFN treatment are required. PMID- 10477399 TI - Cytokines in sepsis due to Candida albicans and in bacterial sepsis. AB - Cytokines, such as tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and tumor necrosis factor-soluble receptor (TNF-sR), and adhesion molecules, e.g. vascular adhesion molecule-1 (VCAM-1) and E-selectin, play an important role in the pathogenesis of bacterial sepsis. Experimental data on cytokine expression during candidaemia are controversial. In this study, plasma concentrations of cytokines and adhesion molecules were compared between patients with sepsis due to Candida albicans and bacterial sepsis. Plasma levels of TNF-alpha, TNF-sR, IL 6, VCAM-1 and E-selectin, were determined in 20 patients with sepsis due to C. albicans, in 20 patients with bacterial sepsis, and in 20 controls on days 1, 7 and 14. On day 1, elevated plasma levels of TNF-alpha, TNF-sR and IL-6 were detected in both sepsis groups compared to controls. On day 1, VCAM-1 levels were higher, and E-selectin levels were lower in patients with Candida sepsis than in patients with bacterial sepsis (p < 0.05). At any time, VCAM-1 levels were significantly greater in patients with Candida sepsis than in patients with bacterial sepsis (p < 0.05). Non-survivors, regardless of the etiology of sepsis, had higher blood levels of IL-6, TNF-sR and E-selectin than survivors. The cytokines, TNF-alpha, IL-6 and TNF-sR, and the adhesion molecules, VCAM-1 and E selectin, are involved in sepsis due to C. albicans as in bacterial sepsis. PMID- 10477400 TI - Tumor necrosis factor-alpha and its soluble receptors in plasma and cerebrospinal fluid of multiple sclerosis patients treated with methylprednisolone. AB - Demyelination is the main pathological feature of multiple sclerosis (MS), a chronic inflammatory disease of the central nervous system. Tumor necrosis factor alpha (TNF-alpha) can cause myelin damage and contribute to MS pathogenesis. We measured plasma and cerebrospinal fluid (CSF) levels of TNF-alpha and its soluble receptors, TNF-sRp55 and TNF-sRp75, in 18 patients with active MS, and in neurological and healthy controls. The same determinations were repeated on plasma and on CSF samples that were collected after the MS patients had ended a six-day treatment with high-dose methylprednisolone (MP). Pre- and post-treatment plasma and CSF TNF-alpha levels, when detectable, and those of TNF-sRp75, did not vary, and were similar to those of controls. CSF TNF-sRp55 levels were higher in acute MS patients than in controls. Post-treatment CSF TNF-sRp55 levels were higher than in the active phase of the disease. The MS patients, who clinically improved, tended to have the highest CSF TNF-sRp55 levels. The increase was due to intrathecal TNF-sRp55 synthesis. Although it is involved in MS pathogenesis, TNF-alpha is not detectable in plasma or in CSF samples from MS patients in various phases of the disease. A better marker of disease activity seems to be CSF TNF-sRp55 levels. The increased CSF levels of TNF-sRp55 in response to MP circumstantially suggest that this receptor could partially account for the beneficial effects of MP in acute MS. PMID- 10477401 TI - A small synthetic molecule capable of preferentially inhibiting the production of the CC chemokine monocyte chemotactic protein-1. AB - Blocking chemokine production or action is a major target for pharmacological intervention in different human diseases. Bindarit (2-methyl-2-[[1-(phenylmethyl) 1H-indazol-3yl]methoxy]propan oic acid) dose-dependently inhibited MCP-1 and TNF alpha production induced in vitro in monocytes by LPS and Candida albicans. It did not affect the production of the cytokines IL-1, IL-6, or the chemokines IL 8, MIP-1alpha and RANTES. In the air pouch model in mice, oral treatment reduced monocyte recruitment and local MCP-1 production, induced by carrageenan or IL-1 injection. In NZB/W mice, a model of lupus nephritis, oral treatment prolonged survival and delayed the onset of proteinuria. The results presented here show that bindarit is a preferential inhibitor of the production of MCP-1 in vitro and in vivo and suggest that its beneficial effects in models of joint and kidney inflammation are related to its anti-MCP-1 action. It is therefore possible to selectively and differentially regulate chemokines by targeting their production with small synthetic molecules. PMID- 10477402 TI - [Non-tuberculous bacterial pleuropneumopathies in children in Abidjan]. AB - We carried out a retrospective study to investigate epidemiological and clinical aspects of patient management in a hospital environment. We studied 90 cases of non-tuberculous bacterial pleuropneumopathies in children, observed over a 5-year period, from January 1 1991 to December 31 1995, at the Pediatric Medical Unit of Yopougon University Hospital. Diagnosis was based on the association of an infectious syndrome with respiratory and digestive disorders, and was confirmed by X ray. Bacteria were isolated from pleural pus and identified in fifty-one cases. Non-tuberculous bacterial pleuropneumopathies accounted for 5.22% of all pneumopathies in children, and the sex ratio for the patients affected was 1.9. Protein and energy malnutrition, anemia, being very young and climatic factors (harmattan and rainy seasons) were identified as major risk factors. The three main bacteria identified were Staphylococcus aureus (21 cases), Streptococcus pneumoniae (19 cases) and Haemophilus influenzae (6 cases). In this series of patients, 20% of the Staphylococcus aureus strains isolated were methicillin resistant. Patients were generally treated with a combination of penicillin A and M and aminoside, with or without a pleural drain. The outcome was favorable in 77.8% of cases, with no severe sequelae detected by X ray. Six of the children died. Bacterial pneumopathies are still common in Sub-Saharan Africa and the patient being very young is indicative of a poor prognosis. PMID- 10477403 TI - [Effect of Onchocerca volvulus infestation on plasma vitamin A concentration in school children in a rural region of Cameroon]. AB - Vitamin A deficiency is known to be associated with immune dysfunction and common childhood infections. However, little is known about the relationship between vitamin A deficiency and onchocerciasis in children. The aim of this study was to determine the prevalence of vitamin A deficiency and to investigate the relationship between vitamin A status and onchocerciasis. A total of 231 children, aged 6 to 15 years, were randomly selected between March 1995 and April 1996 at Yambassa and Balamba (central province of Cameroon). They were examined: we determined their vitamin A status and whether they had onchocerciasis. We diagnosed onchocerciasis by skin biopsy and the detection of antibodies against Onchocerca volvulus in the blood. We found that 101 of the 231 children examined (43.73%) had palpable nodules and/or microfilariae and the remaining 130 (56.27%) had been exposed to the parasite but had no clinical signs of infestation. Some children tested negative for skin microfilariae but positive by ELISA. Thus, 197 (85.28%) children were found to be infested with O. volvulus (group A) and the remaining 34 (14.72%) were found to have been exposed to the parasite but to have no clinical signs of onchocerciasis (group B). Plasma vitamin A concentrations were marginal, with concentrations below 0.7 mumol/l (20 mug/dl) recorded for 82.25% of the subjects. Children with onchocerciasis were more likely to have low vitamin A status. The mean plasma vitamin A concentration of infested children (0.52 +/- 0.14 mumol/l) was significantly lower (p < 0.05) than that of the children exposed but not infested. The parasite, O. volvulus, uses the vitamin A present in host tissues during its development, leading to a decrease in plasma retinol concentration. PMID- 10477404 TI - [Myths concerning malarial transmission among Amazonian Indians and their relation with 2 types of transmission encountered in the Amazonian forest]. AB - Among the Indians Desana's (Tukano amerindians) in the Upper Rio Negro, the interseasonal variation of the malarial fevers were associated with two myths (localised in two distinguishable places). One myth associates the malarial with the rivers which contain "malaria pots". Conception based on an observation of localised water collection in the banks and the rocky rapids ("banks and rocky's fever"). The transmission and the anophelian density present variation between the seasons in relation to the river's level. Another myth associates malarial fevers in the forest, with the song of a frog ("fever's frog") and the flowering and fructification of a tree (Poaqueira sericea Thul.). There is in South America a particular type of forest malaria, known as "Bromelia malaria" and denounced in human and/or simian transmission. This forest malaria is transmitted by the a sub genus anopheles (Kerteszia) which larval breeding are areal in the canopy. The breeding places are found in the forest in the epiphyte bromeliads. To understand this type of transmission, we must take reference to the previous endomological data at the Upper Oyapock Wayapi (Tupi amerindians). This Bromelia malaria could fluctuated according larval density variation, related to washing of epiphytes (end of the rainy season) or to their flowering (end of the dry season). The "fever's frog" myth collected at the Desana's in the Upper Rio Negro can be related to the existence of Bromelia malaria in this amazonian habitat. These myths showed the perfect adaptation of the amerindians to their environment and their complete knowledge of the neotropical forest. PMID- 10477405 TI - [The effect of nutritional management on the mortality of malnourished children, uninfected and infected with the human immunodeficiency virus]. AB - Weight loss is a major complication in children infected with HIV. Very few studies have focused on the nutritional management of malnourished HIV-positive children, particularly in developing countries, although there have been some studies in adults. Therefore, the aim of this retrospective study was to evaluate, as a function of HIV status, the effect of a nutritional rehabilitation program on the mortality of malnourished children in an Ivory Coast nursery. We studied 193 malnourished children over the age of 15 months from January 1 1994 to December 31 1996; 41 % of these children (80 of 193) were HIV-positive. The nutritional rehabilitation program was introduced in 1995. It had a beneficial effect in HIV-negative children because the setback rate (the number of deaths and transfers x 1,000/the number of child-months at risk) decreased significantly over the three years of the study (1994: 130; 1995: 113; 1996: 26; p < 0.05). The rate in HIV-positive children did fall slightly, but this decrease was not statistically significant. These results demonstrate the difficulties involved in the nutritional management of malnourished HIV-positive children. However, recent studies have suggested that nutritional rehabilitation (by mouth) combined with total vitamin and mineral supplementation may be more effective. Given the frequency of malnutrition in HIV-positive children, clinical studies aimed at improving the nutritional management of these children should be a priority in developing countries. PMID- 10477406 TI - [Blood lipid and apolipoprotein concentrations in a healthy urban population of Northern Cameroon: variation with age and sex]. AB - We determined plasma lipids and apolipoproteins A-I and B in 142 subjects from Ngaoundere, in the north of Cameroon, as a means of evaluating atherosclerotic risks. The mean triglyceride concentration was 0.836 +/- 0.174 g/l; mean total cholesterol concentration was 1.276 +/- 0.251; mean HDL cholesterol concentration was 0.162 +/- 0.222; mean apolipoprotein A-I concentration was 0.577 +/- 0.096 and mean apolipoprotein B concentration was 0.836 +/- 0.174. These values differ from those reported for Caucasian populations but are similar to those reported in other studies in Cameroon. All lipid factors except HDL cholesterol concentration increased significantly with age (p < 0. 05). There were also differences between the sexes, with mean triglyceride, LDL cholesterol, HDL cholesterol and apolipoprotein B concentrations significantly higher (p < 0.01) in men than in women. There was no significant difference between the sexes, however, in total cholesterol and apolipoprotein A-I concentrations. Thus advanced age and being male are risk factors for atherosclerosis in this population. PMID- 10477407 TI - [Maternal HIV infection and the anthropometric characteristics of children at birth in Burkina Faso. DITRAME Study Group]. AB - The aim of this study was to investigate the relationship between maternal HIV status and the anthropometric characteristics of children at birth. A cross sectional study was conducted in Bobo-Dioulasso from January 1995 to May 1996. We included all mother-child pairs seen postpartum for whom the HIV status of the mother had been determined at an antenatal visit and anthropometric measurements had been taken for the child at birth. Birth weights below 2,500 g were classed as "low", body lengths of less than 47 cm at birth were described as "small birth size" and head circumferences of less than 33 cm were classed as "small". We included 956 mothers and 956 children in the survey. Low birth-weights were recorded more frequently among children born to HIV-infected mothers than among those born to uninfected mothers (23. 37% versus 15.6%; p = 0.006). Mean birth weight, birth size and head circumference did not differ significantly between the children of HIV-infected and uninfected mothers. In multivariate analysis, HIV infection and primiparity were independently associated with low birth weight. Maternal HIV infection was the only factor associated with small birth size. There was no relationship between head circumference and maternal HIV status. Thus, maternal HIV infection appears to be associated with low birth weight and small birth size. PMID- 10477408 TI - [The value of ultrasound in the diagnosis of thyroid diseases in Mauritania (171 cases)]. AB - We investigated thyroid abnormality in Mauritania by carrying out a prospective ultrasound study of 171 patients. One hundred and thirty four patients underwent thyroid hormone determination (T3, T4 and TSH) and 123 had complementary plain film X rays of the chest and neck. These ultrasound examinations accounted for 3.8% of all ultrasound examinations performed over the study period. Thyroid abnormalities were more frequent in young adults aged 20 to 40 years (67.4%) and in women (83%) in Mauritania. Thyroid diseases affected people from savanna and mountain regions in particular. Diffuse goiters were the most frequent problem (76.3%) and most were multinodular or heterogeneous forms with necrosis or hemorrhage. One case of homogeneous diffuse goiter was found to be an infiltrating vesicular adenocarcinoma with metastatic adenopathy. Nodular goiters were mostly cystic, with septation (51.9%) and could be mistaken for hydatid cysts, which are endemic to the region. Further study of these aspects of thyroid diseases in Mauritania are required. PMID- 10477409 TI - [International bibliographic databases--Current Contents on disk and in FTP format (Internet): presentation and guide]. AB - This paper aims to provide technical and practical advice about finding references using Current Contents on disk (Macintosh or PC) or via the Internet (FTP). Seven editions are published each week. They are all organized in the same way and have the same search engine. The Life Sciences edition, extensively used in medical research, is presented here in detail, as an example. This methodological note explains, in French, how to use this reference database. It is designed to be a practical guide for browsing and searching the database, and particularly for creating search profiles adapted to the needs of researchers. PMID- 10477410 TI - [Methodological assessment of nutritional problems as part of a health care initiative for hospitalized children]. AB - A health care initiative has been set up in a French regional hospital to improve the management of children admitted to the pediatric unit. It involves screening these children, aged between 6 months and 5 years, for nutritional problems. Health care professionals are first trained. They then measure and weigh the children according to a predetermined protocol and calculate an indicator (weight/height ratio) using the EPI-INFO computer program. The medical team is notified if a child is found to be over- or underweight and decisions are taken as to whether further investigations will be carried out. The family are interviewed and the child's eating habits are observed. Nutritional advice is given and the child's doctor is informed. This initiative has been planned and analyzed according to recognized good practice in community programs. All the health care professionals involved in the scheme participated during the year of the study. In total, 1,004 children were admitted to the hospital and 964 were included in the study (97. 8%). Of these, 142 children (14.7%) were included after their discharge from the hospital. Sixty-nine children were identified as being over- or underweight, with 25 (2.6%) underweight and 44 (4.6%) overweight. Sixty-five of these children were identified before their discharge from hospital. The notification rate was only 64.5% and the treatment rate was similar. Efficacy was not assessed. This initiative requires the active participation and awareness of the entire health care team. Improvement should be possible with better computer programs, regular data collection and improved efficiency, which is often not the case when dealing with obesity problems. PMID- 10477411 TI - [Representations of mother to child transmission of AIDS, perception of the risk and health information messages in Burkina Faso]. AB - In Burkina Faso, in rural Mossi areas, popular contemporary representations of the transmission of HIV from mother to child are based on the idea that "the disease is in the blood" and that the fetus is conceived by "mixing the blood" of its parents. Infection of the child is seen as inevitable and systematic and is believed to occur in utero. Maternal milk is thought to have the same potential for infection because "milk is blood" but its role in transmission is seen as secondary, with transmission occurring before birth. However, breast feeding is believed to be responsible for the transmission of the disease in two ways: 1) by infected women transmitting the illness to healthy children via their milk and 2) by healthy women becoming infected by breast feeding infants born to infected mothers. The belief that transmission is systematic and the fear that the child will contaminate others leads to the widely held view within the population that no care should be taken of children born to women with AIDS and that such children should be abandoned and left to die. These representations have recently developed in the population based on preexisting beliefs relating to the physiology and role of blood in the transmission of diseases and the health information that has been distributed. Health information messages are largely responsible for the representations described above. Indeed, the description of the modes of HIV transmission in such messages has been based on the simplified statement of the triad, sex, blood and mother-child, with no indication of the relative risks of transmission for each. Hence, this incomplete information, interpreted in terms of popular conceptions about contagion, has resulted in maximal probability being attributed to each of the listed modes of transmission. Health information messages are the principal means of communicating scientific information to the general population. The stakes associated with the quality and correctness of the information supplied are therefore very high. The notion of the risk of transmission and statement of the level of risk are essential to any explanation of the modes of transmission of HIV. These ideas cannot be neglected because they are essential to the correct understanding of transmission and to the logical management of individual risk. PMID- 10477412 TI - What is the real clinical utility of echocardiography? A prospective observational study. AB - This was a prospective, structured interview to evaluate physician expectations of echocardiography and the subsequent impact on patient care. The setting involved requests for echocardiograms in patients admitted to a tertiary care teaching hospital. Measurements included assessment of the diagnostic and therapeutic implications of echocardiography perceived by the physicians and subsequently reported and confirmed by blinded chart review. From January to May 1997, 542 echocardiograms were performed on 500 inpatients (age 62 +/- 17 years; 56% men). Referring physicians were mainly house staff (83%) and from medical services. The main indications were evaluation of left ventricular function (54%) or valve function (16%). At the time of the request, 89% of physicians believed that echocardiography was needed to guide future investigations or treatment, although in 24% of cases, physicians could not provide details of such. A reported change in treatment occurred in 57% and was validated by chart review in 38%. Changes occurred more frequently in patients in the intensive care unit versus those not in the intensive care unit (54% vs 37%, P =.02) but were similar between attending physicians and house staff (frequency of change 41% vs 39%, P = not significant) and between those with and those without previous echocardiograms (38% vs 39%, P = not significant). The utility of the echocardiogram to influence treatment decisions in hospitalized patients is high, especially in critically ill patients. However, this impact is less than is anticipated at the time of the initial request. Further studies involving more select populations of patients are required. PMID- 10477413 TI - How accurate is visual assessment of synchronicity in myocardial motion? An In vitro study with computer-simulated regional delay in myocardial motion: clinical implications for rest and stress echocardiography studies. AB - Asynchronicity in echocardiographic images is normally assessed visually. No prior quantitative studies have determined the limitations of this approach. To quantify visual recognition of myocardial asynchronicity in echocardiographic images, computer-simulated delay phantom loops were generated from a 3.3 MHz digital image data from a normal left ventricular short-axis heart cycle acquired at 55 frames per second. Six expert observers visually assessed 30 abnormal and 3 normal loops with differing computer-induced delay patterns on 3 occasions and in this optimally simulated environment could recognize only single delays of 89 ms or more. This was improved to 71 ms or more by use of side-by-side (normal versus abnormal) comparative review. Thus visual assessment of clinically important regional delay in rest or stress echo images is limited. PMID- 10477414 TI - Dobutamine stress echocardiography: improved endocardial border definition and wall motion analysis with tissue harmonic imaging. AB - BACKGROUND: We performed a study to determine whether tissue harmonic imaging (THI) facilitates wall motion analysis at rest and whether these benefits extend through the stages of a dobutamine stress echocardiography (DSE) study. We also assessed the impact of THI on the feasibility of DSE in technically difficult patients. Finally we tested the hypothesis that THI by improving endocardial border definition (EBD) could enhance the interobserver agreement between trainees and experienced operators for interpreting DSE studies. METHODS: Twenty unselected patients underwent DSE by standard protocol. Parasternal and apical views were obtained with the use of fundamental mode (FND) and THI at baseline, low dose, and peak stress. Segmental EBD was characterized as 1 to 4 (1 = excellent) and segmental wall motion was characterized as 1 to 4/x (1 = normal, x = unable to interpret) by a consensus of 2 experienced observers. A trainee in stress echocardiography independently scored all segments, and these results were compared with the consensus of the experienced readers. RESULTS: EBD improved with THI in 26 +/- 6.7 of 48 segments per patient (54%, 95% confidence interval [CI] 0.40 to 0.68) and deteriorated with THI in only 2 +/- 2.7 (4%, 95% CI 0 to 0.09). Of the total of 48 segments per patient, a mean of 10 +/- 5.7 (21%, 95% CI 0.10 to 0.31) were of inadequate quality to be interpreted for wall motion on FND, and this changed to 4 +/- 3.4 (6%, 95% CI 0.06 to 0.12) on THI (P <.001). EBD improved in a similar degree in all DSE stages 53%, 54%, and 53% for rest, low dose, and peak stress, respectively. Six of the 20 study patients were deemed unsuitable for DSE on FND, and all were changed to suitable subjects on THI. Of the 205 segments deemed unsuitable for interpretation on FND, 140 (68%) were of the anterior and lateral walls of the LV. Improvement with THI was also more prominent on these walls. The mean coefficient of agreement (kappa) for wall motion analysis was 0.82 +/- 0.14 on FND and improved to 0. 92 +/- 0.09 on THI (P <.001). CONCLUSIONS: THI dramatically improves EBD and the ability to confidently score segmental wall motion. Interobserver agreement is also significantly enhanced. These benefits extend to the peak stage of a DSE study. Routine use of THI may enhance the diagnostic accuracy of DSE and extend its application to technically difficult patients previously deemed unsuitable. PMID- 10477415 TI - A comparison of the interpretation of digitized and videotape recorded echocardiograms. AB - Conventional echocardiograms are typically recorded on videotape and later reviewed and interpreted by a physician. Although videotape recording is an excellent medium for this purpose, it does have several disadvantages, which may be overcome by digital storage. This study compared the diagnostic accuracy of digitized and videotape recorded echocardiograms. Echocardiographic examinations (n = 110) were recorded simulta-neously on videotape and were digitized with a commercially available frame grabber system. Images were transmitted by an Ethernet link to the network-based computer system and compressed with a nondestructive compression algorithm. Images were reviewed on a personal computer. Images were interpreted by 3 observers, and differences in interpretation were documented. There were 274 findings in 110 patients. Exact agreement in interpretation was found in 83%. A major discrepancy occurred in 2%, and a minor discrepancy occurred in 15%. Most discrepancies occurred in the setting of valvular heart disease. When compared with a consensus interpretation, no significant difference was seen in the number of errors between the digital and videotape interpretation. We conclude that the interpretation of a properly recorded digitized echocardiographic examination yields interpretations equivalent to those of videotape recordings. PMID- 10477416 TI - Coronary flow reserve of normal left anterior descending artery in patients with ischemic heart disease: A transesophageal Doppler study. AB - The aim of this study was to investigate the flow reserve of a normal left anterior descending coronary artery (LAD) in patients with coronary artery disease (CAD) of other epicardial vessels by Doppler transesophageal echocardiography (TEE). Thirty-one consecutive patients (age 59 +/- 8 years; 23 men) referred for TEE were considered. Eighteen patients had CAD and a 70% or greater LAD stenosis (group 1); 13 patients had right and/or circumflex CAD (>/=70% stenosis) and normal or minimally diseased LAD (group 2). Ten patients (age 54 +/- 11 years) with normal coronary arteries constituted group 3. Baseline and adenosine (0.160 microg/kg per minute intravenously over 60 minutes) flow velocities in the LAD were measured by pulsed Doppler examination during TEE. Peak and mean systolic and diastolic flow velocities were calculated. Adenosine/baseline peak and mean velocity ratios were used for evaluating blood flow reserve in the LAD. Heart rate and arterial pressure values were similar in the 3 groups at baseline and during adenosine infusion. Baseline and adenosine related flow velocities were comparable in the 3 groups. Peak and mean diastolic velocity ratios were lower in groups 1 and 2 compared with group 3 (peak velocity ratio 1.68 +/- 0.81 and 1.93 +/- 0.35 vs 2.62 +/- 0.32, P <. 05; mean velocity ratio 1.71 +/- 0.86 and 2.01 +/- 0.41 vs 2.84 +/- 0.74, P <.05), whereas no differences were found between groups 1 and 2. No significant differences were found in systolic flow velocity ratios among the 3 groups. Patients with ischemic heart disease have a reduced diastolic flow velocity reserve in the LAD independent from the presence of significant LAD stenosis. Thus the adenosine TEE Doppler study should be considered a screening test for CAD rather than for LAD disease. PMID- 10477417 TI - Echocardiographic parameters of the freestyle stentless bioprosthesis in aortic position: the European experience. AB - The objective of this study was to determine normal Doppler and 2-dimensional characteristics of the Freestyle stentless aortic bioprosthesis. The Freestyle aortic bioprosthesis is a new type of aortic xenograft, and experience is limited. We therefore determined the normal range of echocardiographic and Doppler examinations of this valve. Three hundred thirty-nine consecutive patients with a Freestyle aortic bioprosthesis underwent an echocardiographic and Doppler examination according to a common protocol. Investigations were done within 4 weeks after operation, after 3 to 6 months, and after 1, 2, and 3 years. With a valve size from 19 to 27 mm, mean gradients decreased from 7.9 +/- 5.1 mm Hg at discharge to 5.5 +/- 3. 8 mm Hg after 3 to 6 months (P <.001). Thereafter, gradients remained stable. Effective orifice area 1 year after implantation was 1.59 +/- 0.58 cm(2) for the 21-mm valves, 1.92 +/- 0.74 cm(2) for the 23-mm valves, 2.03 +/- 0.64 cm(2) for the 25-mm valves, and 2.52 +/- 0.72 cm(2) for the 27-mm valves (P <.001). The performance index, the ratio of the measured effective orifice area in the patient divided by the effective orifice area measured in vitro, increased from 67% +/- 20% at discharge to 82% +/- 29% after 1, 2, and 3 years. Performance index was especially very high in the smaller sized valves. After implantation with the subcoronary technique or root-inclusion technique, small cavities could be seen between the native aortic root and the Freestyle valve. Doppler values were evaluated for the Freestyle stentless porcine bioprostheses in the aortic root. Gradients appear to be close to those measured in native valves over a time period of 3 years. PMID- 10477418 TI - The value of assessing pulmonary venous flow velocity for predicting severity of mitral regurgitation: A quantitative assessment integrating left ventricular function. AB - Although alteration in pulmonary venous flow has been reported to relate to mitral regurgitant severity, it is also known to vary with left ventricular (LV) systolic and diastolic dysfunction. There are few data relating pulmonary venous flow to quantitative indexes of mitral regurgitation (MR). The object of this study was to assess quantitatively the accuracy of pulmonary venous flow for predicting MR severity by using transesophageal echocardiographic measurement in patients with variable LV dysfunction. This study consisted of 73 patients undergoing heart surgery with mild to severe MR. Regurgitant orifice area (ROA), regurgitant stroke volume (RSV), and regurgitant fraction (RF) were obtained by quantitative transesophageal echocardiography and proximal isovelocity surface area. Both left and right upper pulmonary venous flow velocities were recorded and their patterns classified by the ratio of systolic to diastolic velocity: normal (>/=1), blunted (<1), and systolic reversal (<0). Twenty-three percent of patients had discordant patterns between the left and right veins. When the most abnormal patterns either in the left or right vein were used for analysis, the ratio of peak systolic to diastolic flow velocity was negatively correlated with ROA (r = -0.74, P <.001), RSV (r = -0.70, P <.001), and RF (r = -0.66, P <.001) calculated by the Doppler thermodilution method; values were r = -0.70, r = 0.67, and r = -0.57, respectively (all P <.001), for indexes calculated by the proximal isovelocity surface area method. The sensitivity, specificity, and predictive values of the reversed pulmonary venous flow pattern for detecting a large ROA (>0.3 cm(2)) were 69%, 98%, and 97%, respectively. The sensitivity, specificity, and predictive values of the normal pulmonary venous flow pattern for detecting a small ROA (<0.3 cm(2)) were 60%, 96%, and 94%, respectively. However, the blunted pattern had low sensitivity (22%), specificity (61%), and predictive values (30%) for detecting ROA of greater than 0.3 cm(2) with significant overlap with the reversed and normal patterns. Among patients with the blunted pattern, the correlation between the systolic to diastolic velocity ratio was worse in those with LV dysfunction (ejection fraction <50%, r = 0.23, P >.05) than in those with normal LV function (r = -0.57, P <.05). Stepwise linear regression analysis showed that the peak systolic to diastolic velocity ratio was independently correlated with RF (P <.001) and effective stroke volume (P <.01), with a multiple correlation coefficient of 0.71 (P <.001). In conclusion, reversed pulmonary venous flow in systole is a highly specific and reliable marker of moderately severe or severe MR with an ROA greater than 0.3 cm(2), whereas the normal pattern accurately predicts mild to moderate MR. Blunted pulmonary venous flow can be seen in all grades of MR with low predictive value for severity of MR, especially in the presence of LV dysfunction. The blunted pulmonary venous flow pattern must therefore be interpreted cautiously in clinical practice as a marker for severity of MR. PMID- 10477419 TI - Accuracy of Doppler methods for estimating peak-to-peak and peak instantaneous gradients across coarctation of the aorta: An In vitro study. AB - Although data exist that address the attempt to correlate noninvasive Doppler derived pressure gradients with invasive catheter pressure gradients in patients with coarctation of the aorta, few data exist about stiffness of the proximal descending aorta (precoarctation) and its relation to these pressure measurements. In this study, an in vitro flow model of a simulated neonatal aorta with a coarctation was developed. Three proximal descending aortas of different stiffnesses were used. The stiffness index of the proximal descending aorta was calculated as beta = ln [systolic pressure/diastolic pressure/(systolic diameter diastolic diameter)]. We evaluated pressure gradients obtained by continuous wave Doppler and standard catheter methods and looked at acceleration of flow velocity determined by pulsed wave Doppler in the 3 precoarctation segments of differing stiffnesses. Pressures in the proximal descending aorta (precoarctation) increased with increasing stiffness, ranging from 105 mm Hg (soft) to greater than 300 mm Hg (stiff). Continuous wave Doppler instantaneous pressure gradients overestimated the catheter instantaneous pressure gradients substantially (mean 41% +/- 19%). The stiffer the precoarctation segment, the more the degree of overestimation: soft, 0% to 63% (= 3.47); medium, 13% to 54% (beta = 4.42); and stiff, 43% to 66% (beta = 5.91). Inclusion of the precoarctation velocity [V1] component in the Bernoulli equation did not significantly improve the correlation or the agreement. An additional observation was that pullback catheter peak-to-peak gradients were higher than simultaneous peak-to-peak gradients. In the stiff aorta, this difference could be greater than 22 mm Hg (>19%). Acceleration of flow velocity toward the coarctation was evident by pulsed wave Doppler interrogation. Increasing the stiffness of the precoarctation segment also increased the degree of acceleration within this proximal segment: soft, 0.4 to 0.8 m/s; medium, 0.5 to 1. 4 m/s; and stiff, 0.7 to 1.5 m/s. These data suggest that increasing stiffness of the proximal descending aorta can alter the continuous wave detected Doppler gradient and although the gradient itself has increased, it may not predict accurately the true severity of the localized, most severely obstructed segment. PMID- 10477420 TI - Multiplane transesophageal echocardiographic acquisition of ascending aortic flow velocities: A comparison with established techniques. AB - Acquisition of ascending aortic flow velocities with monoplane transesophageal echocardiography (TEE) have been problematic because of limitations of available imaging planes and alignment of the Doppler beam with aortic flow. The rotatable imaging array of multiplane TEE (Multi TEE) may provide improved alignment with ascending aortic blood flow. The purpose of this study was to establish the validity of maximal aortic flow velocities (VMax) and velocity time integrals (VTI) obtained by a Multi TEE continuous wave Doppler technique by comparison with those obtained by established echocardiographic techniques, suprasternal Doppler (SSD), and monoplane TEE (Mono TEE). Forty-five patients scheduled for elective surgery were prospectively studied. Multi TEE-obtained VMax and VTI were significantly greater (P <.05), 120 +/- 28.9 cm/s and 25.8 +/- 7 cm, than those obtained by the SSD method, 100.2 +/- 28.6 cm/s and 19.8 +/- 6.8 cm, respectively. Bias analysis revealed that Multi TEE better assessed VMax (mean difference -19.7, SD of the difference of 28 cm/s) and VTI (mean difference -5.9, SD of the difference of 6.4 cm) than the SSD method. Multi TEE exhibited values for VMax 10% or greater than those obtained by SSD in 18 (48. 6%) of 37 patients, and Multi TEE was 10% or greater than SSD in 23 (67%) of 37 patients for VTI determination. Values obtained by Multi TEE and Mono TEE showed close agreement. Multi TEE provides a favorable alignment for continuous wave Doppler interrogation of aortic flow and compared favorably to established techniques. This technique expands the utility of TEE to evaluate aortic valvular function and cardiac performance. PMID- 10477421 TI - Adherence of albunex to an apical left ventricular thrombus. AB - A previously unrecognized left ventricular thrombus developed a distinct echogenic rim after intravenous injection of albumin microbubbles despite no visible opacification of the left-sided chambers. Absence of visible left ventricular opacification suggests a low density of microbubbles crossing the pulmonary vasculature and a high affinity of the microbubbles for the thrombus or its endothelial surface. These findings support previous observations that albumin microbubbles demonstrate transient adherence to abnormal endothelium. PMID- 10477422 TI - Evidence of patent ductus arteriosus and right-to-left shunt by finger pulse oxymetry and Doppler signals of agitated saline in abdominal aorta. AB - The diagnosis of patent ductus arteriosus (PDA) with right-to-left shunt in the adult by using transthoracic echocardiography (TTE) is difficult because of limited windows. We report a case of a 23-year-old woman who was referred for treatment of primary pulmonary hypertension but was found to have a PDA with a right-to-left shunt. On examination she had asymmetric hypoxia of the upper extremities on finger pulse oxymetry. Transthoracic echocardiography revealed a large right atrium and no atrial or ventricular septal wall defects. The diagnosis of PDA with Eisenmenger's syndrome was made after evidence of a right to-left shunt was confirmed by Doppler signals of intravenous agitated saline in the abdominal aorta. This case illustrates the utility of Doppler signals from agitated saline in the abdominal aorta and finger pulse oxymetry, in addition to physical examination, in the diagnosis of PDA with right-to-left shunt. PMID- 10477423 TI - Morphologic analysis of a strand recovered from a prosthetic mitral valve: No evidence of fibrin. AB - We report the first morphologic analysis of a linear mobile structure (strand) detected by transesophageal echocardiography on a bioprosthetic mitral valve and then recovered at surgery. Electron microscopy showed it to consist of a sparsely cellular component, with extracellular amorphous or fibrillary areas. Collagen was largely responsible for the fibrillary appearance. PMID- 10477424 TI - A case for routine screening of coronary artery origins during echocardiography: fortuitous discovery of a life-threatening coronary anomaly. AB - Anomalous origin of the left main coronary artery from the right sinus of Valsalva with retropulmonary course is a rare congenital abnormality. It is associated with a high incidence of sudden cardiac death, particularly among young, athletic individuals. Many of these individuals do not have symptoms before sudden death, and the diagnosis is usually made at postmortem examination. We present a case of a 15-year-old boy who was evaluated for a systolic click with routine 2-dimensional echocardiography. The anomalous coronary artery was serendipitously identified, allowing surgical intervention. Coronary artery origin and proximal course should be visualized on routine echocardiography in the pediatric population. PMID- 10477425 TI - Myxoma of the mitral valve: diagnosis by 2-dimensional and 3-dimensional echocardiography. AB - In this report we describe a 39-year-old patient who had left-sided hemiparesis. In search of a source of embolism, we performed transthoracic echocardiography, which did not show any abnormalities. Transesophageal echocardiography revealed a small tumor of the posterior mitral leaflet. Three-dimensional transesophageal echocardiography was subsequently performed and demonstrated more accurate information about the size, the morphology, and the attachment point of the tumor. Furthermore, the reconstruction provided excellent spatial visualization of the pathomorphology of the mitral valve and was a useful addition for optimal preoperative diagnostic management. The tumor was excised, and histologic examination confirmed the myxomatous character of the tumor. Mitral valve myxomas are rare. This is the first case reported of a mitral valve myxoma being visualized by 3D echocardiography. PMID- 10477426 TI - 6th International HUGO Mutation Database Meeting, March 27, 1999, Brisbane, Australia. AB - The 6th International HUGO Mutation Database Meeting was held on March 27, 1999, in conjunction with the Human Genome Meeting HGM 1999, at the Brisbane Conference and Exhibition Centre, Brisbane, Australia. Meeting highlights are described. PMID- 10477427 TI - Protein misfolding and degradation in genetic diseases. AB - Investigations of genetic diseases such as cystic fibrosis, alpha-1-antitrypsin deficiency, phenylketonuria, mitochondrial acyl-CoA dehydrogenase deficiencies, and many others have shown that enhanced proteolytic degradation of mutant proteins is a common molecular pathological mechanism. Detailed studies of the fate of mutant proteins in some of these diseases have revealed that impaired or aberrant folding of mutant polypeptides typically results in prolonged interaction with molecular chaperones and degradation by intracellular proteases before the functional conformation is acquired. This appears to be the case for many missense mutations and short in-frame deletions or insertions that represent a major fraction of the mutations detected in genetic diseases. In some diseases, or under some circumstances, the degradation system is not efficient. Instead, aberrant folding leads to accumulation of protein aggregates that damage the cell. Mechanisms by which misfolded proteins are selected for degradation have first been delineated for the endoplasmatic reticulum; this process has been termed "protein quality control." Similar mechanisms appear to be operative in all cellular compartments in which proteins fold. Within the context of genetic diseases, we review knowledge on the molecular processes underlying protein quality control in the various subcellular compartments. The important impact of such systems for variability of the expression of genetic deficiencies is emphasised. PMID- 10477428 TI - Molecular basis of the neuronal ceroid lipofuscinoses: mutations in CLN1, CLN2, CLN3, and CLN5. AB - The neuronal ceroid lipofuscinoses (NCLs), also referred to as Batten disease, are a group of neurodegenerative disorders characterised by the accumulation of an autofluorescent lipopigment in many cell types. Different NCL types are distinguished according to age of onset, clinical phenotype, ultrastructural characterisation of the storage material, and chromosomal location of the disease gene. At least eight genes underlie the NCLs, of which four have been isolated and mutations characterised: CLN1, CLN2, CLN3, CLN5. Two of these genes encode lysosomal enzymes, and two encode transmembrane proteins, at least one of which is likely to be in the lysosomal membrane. The basic defect in the NCLs appears to be associated with lysosomal function. PMID- 10477429 TI - TP53 mutation and haplotype analysis of two large African American families. AB - Two large apparently unrelated African American families with a high incidence of breast cancer and other tumors characteristic of Li-Fraumeni breast sarcoma cancer family syndrome were studied. Mutation screening revealed that in both families the affected members carried a germline mutation of the TP53 gene at codon 133 (ATG--> ACG, M133T). In order to determine whether an ancestral haplotype was shared by these two families, polymorphic markers within and flanking the TP53 gene were studied. Haplotype analysis using five markers revealed an identical haplotype shared by the two families. Loss of heterozygosity at the TP53 locus in the probands' tumor tissues from each family was observed; in each case, the retained allele carried the common haplotype. The frequency of this haplotype in the general African American population is <0.003. This unique haplotype, combined with the rare TP53 mutation, suggests that these African American families share a common ancestry. This finding suggests that other African Americans may be carriers of this mutation and thus may be at risk of early-onset breast cancer or other cancers characteristic of the Li-Fraumeni breast sarcoma cancer family syndrome. The finding of recurring mutations in African Americans may facilitate carrier screening and identification in this population. PMID- 10477430 TI - Screening for mutations in the uroporphyrinogen decarboxylase gene using denaturing gradient gel electrophoresis. Identification and characterization of six novel mutations associated with familial PCT. AB - The two porphyrias, familial porphyria cutanea tarda (fPCT) and hepatoerythropoietic porphyria (HEP), are associated with mutations in the gene encoding the enzyme uroporphyrinogen decarboxylase (UROD). Several mutations, most of which are private, have been identified in HEP and fPCT patients, confirming the heterogeneity of the underlying genetic defects of these diseases. We have established a denaturing gradient gel electrophoresis (DGGE) assay for mutation detection in the UROD gene, enabling the simultaneous screening for known and unknown mutations. The established assay has proved able to detect the underlying UROD mutation in 10 previously characterized DNA samples as well as a new mutation in each of six previously unexamined PCT patients. The six novel UROD mutations comprise three missense mutations (M01T, F229L, and M324T), two splice mutations (IVS3-2A-->T and IVS5-2A-->G) leading to exon skipping, and a 2 bp deletion (415-416delTA) resulting in a frameshift and the introduction of a premature stop codon. Heterologous expression and enzymatic studies of the mutant proteins demonstrate that the three mutations leading to shortening or truncation of the UROD protein have no residual catalytic activity, whereas the two missense mutants retained some residual activity. Furthermore, the missense mutants exhibited a considerable increase in thermolability. The six new mutations bring to a total of 29 the number of disease-related mutations in the UROD gene. The DGGE assay presented greatly improves the genetic diagnosis of fPCT and HEP, thereby facilitating the detection of familial UROD deficient patients as well as the discrimination between familial and sporadic PCT cases. PMID- 10477431 TI - Molecular analyses of the vasopressin type 2 receptor and aquaporin-2 genes in Brazilian kindreds with nephrogenic diabetes insipidus. AB - Nephrogenic diabetes insipidus (NDI) is associated with germline mutations in two genes: vasopressin receptor type 2 (V2(R)) in X-linked NDI, and the water channel aquaporin-2, in autosomal-recessive disease. Genetic heterogeneity is further emphasized by reports of phenotypically abnormal individuals with normal structural genes. We analyzed both genes in five Brazilian families and the aquaporin-2 gene in two Swedish families with clinical and laboratory diagnosis of NDI, by a combination of denaturing gradient gel electrophoresis (DGGE) and direct DNA sequencing. A novel polymorphism in the aquaporin-2 gene (S167S), but no disease-associated mutations in any tested individual from all seven families, was detected. In two Brazilian families, frameshift mutations were detected in the V2(R) gene: one leading to a premature stop after codon 36 and the other to a longer peptide (462 aa instead of the 373 aa wild-type protein). In two other Brazilian families, probable disease-associated missense mutations were detected: an alanine to proline at codon 163 (A163P) and an asparagine to aspartic acid at codon 85 (D85N). In one Brazilian family, both genes were structurally normal and the aquaporin-2 gene was also normal in the two Swedish kindreds. This report further extends the mutational spectrum of NDI and suggests that there are other mutational or epigenetic events inactivating the two known genes or even novel genes that underlie NDI. PMID- 10477432 TI - Identification of 12 novel mutations and two new polymorphisms in the arylsulfatase A gene: haplotype and genotype-phenotype correlation studies in Spanish metachromatic leukodystrophy patients. AB - Arylsulfatase A (ARSA) deficiency is the main cause of metachromatic leukodystrophy (MLD), a lysosomal disorder with no specific treatment. In view of the importance of genetic counseling, analyses of mutations and polymorphisms, including the ARSA pseudodeficiency allele, were carried out in 18 unrelated Spanish MLD patients. A systematic search allowed us to identify 100% of the alleles involving 17 different mutations, 12 of which are novel: G32S, L68P, R84W, P94A, G99V, P136S, W193X, H227Y, R288H, G308D, T327I, and IVS6-12C-->G. Two new polymorphisms, 2033C>T and 2059C>T, were identified in intron 6 which, in combination with two polymorphisms previously described (2161C>G and 2213C>G), gave rise to four different haplotypes in the control population. In addition, we also studied polymorphism 842G>T. Linkage disequilibrium was detected between mutations IVS2+1G-->A, D255H, and T327I and specific haplotypes, suggesting a unique origin for these mutations. Moreover, mutation T327I was always associated with the T allele of the new rare variant A210A (893C>T). The distribution of mutation D255H (frequency 19.4%) among patients with different MLD clinical presentation revealed a clear genotype-phenotype correlation paralleling that reported for mutation IVS2+1G-->A (frequency 25%). Among the novel mutations, only P136S and R288H occurred on a background of the ARSA pseudodeficiency allele. Screening 182 normal chromosomes identified a frequency of 8.8% of this allele; moreover, we identified two unrelated subjects with the polyA- mutation in the absence of the N350S mutation, and this infrequent haplotype reinforced the heterogeneity of conditions with ARSA deficiency. PMID- 10477434 TI - Molecular basis of late-life globoid cell leukodystrophy. AB - Globoid cell leukodystrophy is an autosomal recessive inherited disease caused by deficiency of the lysosomal enzyme galactocerebrosidase (GALC). Although the severe, rapidly progressing infantile form is the most common, late-onset forms have been described. We investigated the molecular basis of GALC deficiency in a patient with a late-life mild form of globoid cell leukodystrophy who survived into the eighth decade. Since material suitable for mutation analysis was no longer available from the proband, her GALC genotype was reconstructed by analyzing this gene in her six obligate carrier offspring. One allele contained the mutation 809G>A (G270D) in the 1637C background, while the other allele contained three sequence variants: 1609G>A (G537R), 1873G>A (A625T), and 1650T>A (V550V) in the 1637T background. These mutations were confirmed in the proband's genomic DNA isolated from a sural nerve biopsy. Expression studies indicated that the G537R is a disease-causing mutation, as it resulted in no GALC activity, either alone or together with the A625T. This A625T sequence variant did not affect the enzyme activity, at least when expressed in the 1637T background. The mild clinical phenotype was likely to be associated with the 809G>A, since residual GALC activity, about 17% of the control activity, was detected in the expression studies of this mutation. This mutation has been found in several other patients with late-onset GLD. PMID- 10477433 TI - E-cadherin germline mutations define an inherited cancer syndrome dominated by diffuse gastric cancer. AB - To extend earlier observations of germline E-cadherin mutations in kindreds with an inherited susceptibility to diffuse gastric cancer, we searched for germline E cadherin mutations in five further families affected predominantly by diffuse gastric cancer and one family with a history of diffuse gastric cancer and early onset breast cancer. Heterozygous inactivating mutations were found in the E cadherin gene in each of these families. No mutation hotspots were identified. These results demonstrate that germline mutation of the E-cadherin gene is a common cause of hereditary diffuse gastric cancer and suggest a role for these mutations in the incidence of breast cancer. PMID- 10477435 TI - Determination of the carrier frequency of the common GJB2 (connexin-26) 35delG mutation in the Belgian population using an easy and reliable screening method. AB - Mutations in the gene GJB2, encoding the gap-junction protein connexin-26, have been shown to be a major cause of nonsyndromic recessive deafness (NSRD). A single mutation in the GJB2 gene accounts for the majority of NSRD in many different populations. This mutation represents a deletion of a guanine within a stretch of six Gs between nucleotide positions +30 and +35 of the GJB2 cDNA (35delG). Molecular detection of the 35delG mutation is usually performed by direct sequencing analysis of PCR products, or by allele-specific PCR analysis. To screen for this mutation, we developed an easier and more reliable method, based on the principle of PCR-mediated site-directed mutagenesis (PSDM), followed by a BsiYI digestion. We tested 360 unrelated unaffected Belgian individuals for heterozygosity of the 35delG mutation and found a carrier frequency of 1 in 40 (95% CI, 1 in 30 to 1 in 60). As our new screening method is simple and reliable in use, and detects a mutation responsible for a significant part of NSRD, it may find widespread use in DNA diagnostics. PMID- 10477436 TI - Deep intronic mutations are rarely a cause of hemophilia B. PMID- 10477437 TI - A common 3-bp deletion in the DYT1 gene in Russian families with early-onset torsion dystonia. AB - Hereditary torsion dystonia represent a clinically and genetically heterogeneous group of movement disorders. The most severe and frequent form of hereditary torsion dystonia is early-onset generalized dystonia, DYT1. The DYT1 gene (Ozelius et al., 1997) encodes an ATP-binding protein torsin A. A unique 3-bp deletion (GAG) was found in the heterozygous state in almost all patients with early-onset dystonia from different populations. We observed 39 patients with early-onset generalized torsion dystonia belonging to 22 families from Russia. Seven families were of Ashkenazi Jewish (AJ) ethnic background, and other patients originated from the Slavonic population of Russia. The GAG deletion was identified in 24 affected persons from 15 families (68.2% of the families studied). In all the 7 families of AJ origin the disease was found to be caused by the deletion. In Slavs, the deletion was identified in 8 of 15 families (53%). In two deletion-positive families we observed the co-occurrence of typical early onset generalized dystonia and atypical phenotypes-either isolated postural hand tremor or stutter. PMID- 10477438 TI - Racial differences in the frequencies of cardiac beta(1)-adrenergic receptor polymorphisms: analysis of c145A>G and c1165G>C. PMID- 10477439 TI - A noval nonsense mutation (Y849X) in the CFTR gene of a CF patient from southern Italy. PMID- 10477440 TI - Urban air particulate inhalation alters pulmonary function and induces pulmonary inflammation in a rodent model of chronic bronchitis. AB - Epidemiological studies have reported increased morbidity in human populations following inhalation of elevated levels of urban particulate matter. These responses are especially prevalent in populations with chronic obstructive pulmonary diseases, including chronic bronchitis. Toxicological studies have reported altered pulmonary function and increased pulmonary inflammation following particulate inhalation in the laboratory setting. However, most of these studies have utilized artificial particles that may not accurately mimic outdoor air pollutant conditions. Few studies have utilized actual urban air particle samples in inhalation studies. In the present study, the effects of inhaled concentrated urban air particulates on pulmonary function and pulmonary inflammation are addressed. Normal rats and rats with chronic bronchitis induced by approximately 200 ppm SO(2) for 6 wk were subsequently subjected to filtered air or concentrated air particles (CAPs). Twelve rats per group in 4 groups (48 rats total) were exposed for 5 h/day for 3 consecutive days. The CAPs aerosol levels were 206, 733, and 607 microg/m(3) (MMAD = 0.18 microm, sigma(g) = 2.9) on days 1, 2, and 3, respectively. Following the final day of exposure, pulmonary function parameters, including peak expiratory flow (PEF), tidal volume (TV), respiratory frequency (RF), and minute volume (MV), were measured and compared to preexposure baseline levels. Twenty-four hours following the final day of exposure, bronchoalveolar lavage was performed for total cell counts, differential cell counts, and total lavage protein levels. Pulmonary responses to CAPs in chronic bronchitic animals indicated a significant increase in tidal volume as well as peak expiratory flow. In CAPs-exposed animals without underlying bronchitis, significantly increased tidal volume was observed. Significant pulmonary inflammation was observed in the CAPs-exposed animals, particularly those with chronic bronchitis. Significant increases in neutrophils, lymphocytes, and total lavage protein were observed. These results suggest two distinct mechanistic responses to inhaled particles: a stress-type pulmonary function response marked by increases in flow and volume, that is, deeper breathing; and acute pulmonary inflammation marked by cellular influx, particularly neutrophils. From these data it is concluded that inhaled urban air particles alter pulmonary breathing parameters and increase pulmonary inflammation. PMID- 10477441 TI - A comparison of biomarkers of ozone exposure in human plasma, nasal lavage, and sputum. AB - We examined ozone-induced upper and lower airway inflammatory responses and the concentrations of hydroxylated salicylate metabolites using nasal lavage fluid and induced sputum, in order to identify noninvasive and sensitive biomarkers for ozone exposure and effects. A time course for plasma concentration of 2, 3 dihydroxybenzoic acid (2,3-DHBA, a salicylate metabolite and an indicator for hydroxyl radical) in response to 0.12 ppm ozone was also studied. Healthy, young, nonsmoking volunteers were given acetylsalicylic acid (ASA, 975 mg) or placebo orally. Subjects were exposed to ozone (0.12 or 0.4 ppm) or filtered air in an environmental chamber for 2 h, while performing intermittent exercise. Blood was collected hourly over a 4-h period. After exposure, nasal lavage fluid was collected, and sputum was induced using hypertonic saline. Results show that in sputum the percentage of neutrophils was significantly higher after the subjects were exposed to 0.4 ppm ozone (p<.05) than after they were exposed to filtered air or 0.12 ppm ozone. The absolute number and the percentage of macrophages were significantly lower at 0.4 ppm ozone than for filtered air control or 0.12 ppm ozone. The percentage of lymphocytes in sputum was also significantly lower at 0.4 ppm ozone than for filtered air control or 0.12 ppm ozone. The sputum cellular responses to ozone were not significantly altered by ASA treatment. In nasal lavage, cell counts and differentials did not change significantly after exposure to ozone in comparison to filtered air control. The cellular data indicate an acute inflammation developed during ozone exposure in the lower respiratory tract. The concentrations of total protein and interleukin-8 and the activity of N-acetyl-beta-D-glucosaminidase (a lysosomal enzyme) in nasal lavage and sputum did not change significantly following exposure to ozone in comparison to filtered air control. Plasma 2,3-DHBA concentration increased significantly following exposure to 0.12 ppm ozone in an exposure-dependent temporal pattern. Salicylate metabolites in nasal lavage fluid and sputum did not increase significantly following exposure to ozone. There was a marked variation of 2,3 DHBA concentrations in airway fluids. Data suggest that plasma 2,3-DHBA is a sensitive marker indicating acute ozone exposure, even at an ozone concentration that causes minimal observable airway effects in healthy subjects. PMID- 10477442 TI - Cytokine profile of human bronchoalveolar macrophages and bronchial epithelial cells in response to inhalation particles of the cyclosporine derivative IMM 125. AB - Administration of antiasthmatic drugs in the form of inhalation particles may alter the cytokine network in the airways, independently of their pharmacological actions. Changes induced by drugs not well tolerated may potentially contribute to the immunopathology of the disease, a strongly undesirable effect. In this study, cell viability assays and characterization of the cellular profile of cytokines and chemokines were performed in order to investigate the response of human bronchoalveolar macrophages and bronchial epithelial cells in culture to inhalation particles of the cyclosporine derivative IMM 125. Interleukin 1beta (IL-1beta), tumor necrosis factor alpha (TNFalpha), and IL-8 were assayed by enzyme-linked immunosorbent assay (ELISA) in the supernatants of bronchoalveolar macrophages, and RANTES, granulocyte--macrophage colony-stimulating factor (GM CSF), and IL-8 in those of bronchial epithelial cells. Cells were studied both under basal and stimulated conditions (lipopolysaccharide and TNFalpha were used for activating macrophages and epithelial cells, respectively). The immunosuppressant FK 506 and the glucocorticoid Budesonide served as comparison. IMM 125 did not affect cell viability (except at high concentrations and long time periods). Moreover, IMM 125 did not induce an increase in the secretion of any of the cytokines and chemokines measured with respect to nontreated cells, except for a slight increase in IL-8, an effect that was also observed for FK 506, Budesonide, and inert latex particles, and was therefore regarded as nonspecific. Furthermore, IMM 125 significantly decreased the secretion of TNFalpha, IL-1beta by macrophages, and GM-CSF by epithelial cells, suggesting an antiinflammatory potential. In conclusion, the present in vitro results point to a good tolerance of human airways to IMM 125 inhalation particles. PMID- 10477443 TI - A 2-year inhalation study of phosphine in rats. AB - Phosphine is a highly toxic gas used as a fumigant, a dopant in semiconductor manufacturing, and in the production of organophosphines. In a chronic toxicity and oncogenicity study of phosphine, 60 male and female F344 rats per group were exposed via whole-body inhalation for 6 h/day, 5 days/wk for up to 104 wk to mean concentrations of 0, 0.3, 1, or 3 ppm phosphine. Three parts per million was considered the maximum exposure level because of lethality seen at higher exposure levels in previous repeat dose studies. Ten rats per sex per group were sacrificed after 52 wk of exposure. Survivors were sacrificed after 104 wk of exposure. There were no phosphine-related effects seen on clinical observations, body weight, food consumption, hematology, clinical chemistry, urinalysis, or ophthalmology. There were no phosphine-related macroscopic findings or effect on absolute or relative organ weights. No histomorphologic alterations attributable to phosphine exposure were seen. In conclusion, under the conditions of this study, there were no treatment-related changes suggestive of a toxic or carcinogenic effect seen in rats following 52 wk or 2 yr of whole-body inhalation exposure to 0.3, 1, or 3 ppm phosphine. PMID- 10477445 TI - Time-course analysis of type II cell hyperplasia and alveolar bronchiolization in rats treated with different particulates. AB - In this study, we compared the morphological reaction patterns in rat lungs following a single intratracheal instillation of 20 mg quartz, 20 mg coal mine dust (15.3% quartz), or 25 mg talc. Control animals received a single dose of 0.5 ml saline solution intratracheally. Investigations by light microscopy, morphometry, and DNA image cytometry were carried out 3, 6, 12, and 18 mo after dust administration. During the investigation period, we observed a temporary increase in the number, area, and proliferative activity of the type II cells, which differed in intensity among the three dusts. After 18 mo, however, type II cells in treated animals did not differ from control animals. On the other hand, the expansion of a multifocal alveolar bronchiolization as putative preneoplastic lesion had progressed enormously by the end of the test (1-3% of the investigated lung area). Consistent with this, the proliferative activity of the epithelial cells in terminal bronchi of the coal mine dust- and quartz-treated animals was enhanced by the end of the 18-mo investigation period, while the reaction to talc was minimal (0.2% of totally investigated lung area). Our data suggest that in bronchiolo-alveolar regions, especially in the epithelium of terminal bronchi, there is an overshoot regeneration after cell damage that leads to an alveolar bronchiolization. PMID- 10477444 TI - Short-term inflammatory responses following intratracheal instillation of fine and ultrafine carbon black in rats. AB - Ultrafine carbon black (ufCB) 14 nm in diameter and fine carbon black (CB) 260 nm in diameter were instilled intratracheally in rats at mass of 125 microg, and the bronchoalveolar lavage (BAL) profile at 6 h was assessed. UfCB generated a 50% neutrophil alveolitis 6 h after intratracheal instillation compared to CB, which showed similar activity to the phosphate-buffered saline (PBS) vehicle control. UfCB instillation also produced a marked increase in lactate dehydrogenase (LDH) levels in BAL fluid, which was associated with increased epithelial permeability measured as total protein. In contrast, CB had much less of an effect in increasing BAL protein. Although both particle types caused a decrease in glutathione (GSH) in lung tissue compared to control, the greatest depletion was seen in ufCB-treated animals. To investigate time response, bronchoalveolar lavage was carried out at 6 h, 24 h, and 7 days after a single 125-microg instillation of ufCB. Neutrophil influx was relatively persistent and was still maintained 7 days later. Tumor necrosis factor (TNF) production by BAL leukocytes increased gradually postinstillation, whereas NO production became significantly higher at 24 h after instillation and remained at raised levels up to 7 days. Higher doses of CB caused more inflammation than the ufCB. Thus, in the instillation model, a localized dose of particle over a certain level causes the particle mass to dominate the response, rather than the surface area. In contrast to the effect of CB, which showed a dose-related increasing inflammatory response, ufCB at the highest dose caused less of a neutrophil influx than at the lower dose. Six hours after intratracheal instillation, the threshold dose for neutrophil influx occurred at 50 microg. Calculation of surface area of particles instilled suggested that this was likely to be an overload-inducing dose of particles, as gauged from recent experiments with inhaled particles. In summary, this study provides evidence in a rat instillation model that ufCB has greater ability than CB to produce lung inflammation and oxidant stress at a relatively low dose of 125 microg. At high doses, however, BAL is not a reliable indicator of pulmonary response, since the overall response seems to scale to mass or volume of instilled particulate without an influence of surface area. PMID- 10477446 TI - BHA diet enhances the survival of mice exposed to phosgene: the effect of BHA on glutathione levels in the lung. AB - Phosgene-induced pulmonary edema formation has been under investigation for many years. One mechanism of protection may involve the use of antioxidants. Previously, it has been shown that butylated hydroxyanisole (BHA) treatment can enhance glutathione (GSH) levels. The present study focused on dietary supplementation in mice using BHA, a phenolic compound used in food preservation. Three groups of male CD-1 mice were studied: group 1, control animals fed with Purina rodent chow 5002; group 2, fed 0.75% BHA (w/w) in 5002; and group 3, fed 1.5% BHA (w/w) in 5002. Mice were fed for 22 days. On day 23 mice were exposed to 32 mg/m(3) phosgene for 20 min in a whole-body exposure chamber. Survival rate (SR) and odds ratio (OR) were determined at 12 and 24 h. In mice that died within 12 h, the lungs were removed immediately and lung wet weights (WW), dry weights (DW), lung wet weight/body weight ratio (LWW/BW), and lung tissue total glutathione (GSH) were assessed. For 12-h data, 6 mice from the 1.5% BHA group were sacrificed for lung tissue measurements. The SR for 0.75% BHA was 80% at 12 h and 55% at 24 h, compared with 36% and 23%, respectively, for controls. For 1.5% BHA, the 12- and 24-h SR were 100% and 92%, respectively. Odds ratios of 6.9 for 0.75% BHA and 46.6 for 1.5% BHA at 12 h and 4.0 and 42 for 0. 75% and 1.5% BHA, respectively, at 24 h were significantly (chi2) higher than control diet phosgene-exposed mice. Dietary pretreatment with 0.75% and 1.5% BHA significantly enhanced lung tissue GSH, 1.8-fold (p < or =.01) and 5.8-fold (p < or =.01), respectively, compared with phosgene-exposed control diet. Both BHA-supplemented diets significantly reduced WW. Only 1.5% BHA reduced DW, a measure of lung hyperaggregation. and LWW/BW compared with control diet. In air-exposed controls, BHA induced a dose-responsive decrease in WW, DW, LWW/BW ratio, and GSH. In conclusion, dietary pretreatment with BHA at the two dose levels reduced lung edema and lethality by enhancing lung tissue GSH in mice exposed to phosgene. PMID- 10477447 TI - Langerhans' cell histiocytosis in adults: another orphan disease? PMID- 10477448 TI - Clonogenic potential and phenotypic analysis of CD34+ cells mobilized by different chemotherapy regimens. AB - BACKGROUND AND OBJECTIVE: Since limited data concerning quantitative and qualitative differences of CD34+ cells collected after different mobilization schedules are available, we investigated phenotype, proliferative capacity and primitive progenitor cell content of CD34+ cells mobilized with four different regimens. DESIGN AND METHODS: The number, phenotype, and progenitor cell content of CD34+ cells were investigated in 46 patients mobilized with cyclophosphamide (CY) 7 g/m(2) plus granulocyte colony-stimulating factor (G-CSF, 5 microg/kg) (CY7+G-CSF) (n=16), CY 4 g/m(2) plus G-CSF (CY4+G-CSF) (n=8), IVE [ifosphamide (2.5 g/m(2) for 3 d), etoposide (150 mg/m(2) for 3 d), epirubicin (100 mg/m(2) on day 1)] plus G-CSF (IVE+G-CSF) (n=9), or G-CSF (10 microg/kg) alone (n=13). RESULTS: The number of CD34+ cells collected per liter of processed blood was significantly higher in the CY7+G-CSF group than in the CY4+G-CSF and G-CSF groups (p 60 years; AIDA GIMEMA for APL patients. All patients submitted to the AML10 and AML13 protocols and those patients submitted to the AIDA protocol with difficult peripheral vein access had a central venous catheter (CVC) sited. Patients treated as in-patients were discharged at the end of consolidation chemotherapy provided they were in a good clinical condition. They were routinely evaluated on an out-patient basis twice weekly. In the event of any complication they were referred to the Emergency Unit of our Department dedicated to out-patients with hematologic diseases. RESULTS: One hundred and eleven patients with AML were eligible for intensive chemotherapy. After achievement of complete remission they received a total of 133 consolidation courses and in 127 instances they were followed on an out-patient basis during the aplastic phase. There were 69 cases (54%) of rehospitalization, 68 because of fever and only one because of severe anemia. Rehospitalization occurred in 90%,70% and 38% of courses in AML10, AML13 and AIDA protocols, respectively. Only one patient died: the cause of death was a brain hemorrhage. Coagulase negative staphylococci and viridans streptococci were the organisms most frequently isolated from blood. Most coagulase negative staphylococci were isolated in patients submitted to AML10 and AML13 protocols, who had an indwelling CVC. Empiric once-a-day antibacterial therapy with ceftriaxone and amikacin was effective in 75% of the cases and made early discharge possible in 28% of the cases with antibiotic therapy continued in an out-patient setting. Overall, patients were managed out of the hospital for 66% of the period of post-consolidation neutropenia (77%, 48% and 50% of the post consolidation neutropenia period in patients treated with AIDA, AML10 and AML13 protocols, respectively). INTERPRETATION AND CONCLUSIONS: Thanks to the availability of an emergency unit specifically dedicated to out-patients with hematologic diseases, selected out-hospital management of AML patients during post-consolidation cytopenia is a feasible, well accepted and cost-saving option, and can contribute to lower the risk of developing severe nosocomial infections. The empiric therapy with once-a-day ceftriaxone plus amikacin was effective, with the exception of staphylococcal infections, and made it possible to discharge patients early to continue treatment in an out-patient setting. PMID- 10477456 TI - Community respiratory virus infections in patients with hematologic malignancies. AB - BACKGROUND AND OBJECTIVE: The main difficulty of PCR-based clonality studies for B-cell lymphoproliferative disorders (B-LPD) is discrimination between monoclonal and polyclonal PCR products, especially when there is a high background of polyclonal B cells in the tumor sample. Actually, PCR-based methods for clonality assessment require additional analysis of the PCR products in order to discern between monoclonal and polyclonal samples. Heteroduplex analysis represents an attractive approach since it is easy to perform and avoids the use of radioactive substrates or expensive equipment. DESIGN AND METHODS: We studied the sensitivity and specificity of heteroduplex PCR analysis for monoclonal detection in samples from 90 B-cell non Hodgkin's lymphoma (B-NHL) patients and in 28 individuals without neoplastic B-cell disorders (negative controls). Furthermore, in 42 B-NHL and in the same 28 negative controls, we compared heteroduplex analysis vs the classical PCR technique. We also compared ethidium bromide (EtBr) vs. silver nitrate (AgNO(3)) staining as well as agarose vs. polyacrylamide gel electrophoresis (PAGE). RESULTS: Using two pair consensus primers sited at VH (FR3 and FR2) and at JH, 91% of B-NHL samples displayed monoclonal products after heteroduplex PCR analysis using PAGE and AgNO(3) staining. Moreover, no polyclonal sample showed a monoclonal PCR product. By contrast, false positive results were obtained when using agarose (5/28) and PAGE without heteroduplex analysis: 2/28 and 8/28 with EtBr and AgNO(3) staining, respectively. In addition, false negative results only appeared with EtBr staining: 13/42 in agarose, 4/42 in PAGE without heteroduplex analysis and 7/42 in PAGE after heteroduplex analysis. INTERPRETATION AND CONCLUSIONS: We conclude that AgNO(3) stained PAGE after heteroduplex analysis is the most suitable strategy for detecting monoclonal rearrangements in B-NHL samples because it does not produce false-positive results and the risk of false-negative results is very low. PMID- 10477457 TI - C677T substitution in the methylenetetrahydrofolate reductase gene as a risk factor for venous thrombosis and arterial disease in selected patients. AB - BACKGROUND AND OBJECTIVE: Hyperhomocysteinemia, due to a combination of genetic and environmental factors, is considered to be a risk factor for vascular disease. Individuals with the thermolabile variant of methylenetetrahydrofolate reductase (MTHFR), due to homozygous C677T MTHFR gene mutation, have significantly raised plasma levels of homocysteine and may be at increased risk of vascular disease. However, it is still controversial a direct association between C677T homozygosity and the occurrence of vascular disease is still controversial. DESIGN AND METHODS: To clarify the contribution of C677T MTHFR mutation in arterial occlusive disease (AOD) or venous thromboembolism (VTE), we performed a case-controlled study including 160 cases with AOD and 180 cases with VTE attending our referral center and compared them with 200 matched healthy controls. MTHFR gene mutation was evaluated by PCR and odds ratios (OR) and the 95% confidence intervals (CI) were used to estimate the risk for venous or arterial thrombosis. RESULTS: There was a high prevalence of homozygotes for the mutated MTHFR allele among the whole group of cases with arterial disease (OR = 2.35, p = 0.001). Considering the AOD cases with and those without associated risk factors for arterial disease separately the difference remained significant only in the latter group (p = 0.168 and P<0.001 respectively). In contrast, the prevalence of mutated homozygotes among the whole group of cases with VTE was not significantly different from that in the control group (OR = 1.67; p = 0.070). Excluding VTE cases with inherited thrombophilia or with circumstantial risk situations the value increased in both subgroups (OR = 2.26; p = 0.006 and OR = 2.03; p = 0.033 respectively). Considering only VTE cases with neither inherited thrombophilia nor circumstantial risk situations the risk increased further (OR = 2.57; p = 0.017). INTERPRETATION AND CONCLUSIONS: These data suggest that in selected patients homozygosity for the MTHFR mutation increases the risk of both arterial and venous thromboses and that differences in selection criteria for the patient group may be responsible in part for the controversial association of the MTHFR mutation and vascular disease. PMID- 10477458 TI - IgG reactivity to phospholipid-bound beta(2)-glycoprotein I is the main determinant of the fraction of lupus anticoagulant activity quenched by addition of hexagonal (II) phase phospholipid in patients with the clinical suspicion of antiphospholipid-antibody syndrome. AB - BACKGROUND AND OBJECTIVE: Autoantibodies to beta(2)-glycoprotein I (beta(2)-GPI) and/or prothrombin (FII) have been involved in the expression of lupus anticoagulant (LA) activity, an in vitro phenomenon associated with an increased risk of arterial and/or venous thromboembolic events. However, LA activity sustained by anti-FII antibodies has a much weaker association with thrombosis than LA activity sustained by anti-beta(2)-GPI antibodies. Because assays aimed at detecting LA activity are now commercially available, we evaluated the relative sensitivity to anti-FII and anti-beta(2)-GPI antibodies of a commercial LA assay in a consecutive series of patients with the clinical suspicion of anti phospholipid antibody (APA) syndrome. DESIGN AND METHODS: One hundred and ten consecutive patients with the clinical suspicion of APA syndrome (primary in 39) and 36 healthy controls were evaluated for the presence of LA activity (LA, Staclot, Stago), anticardiolipin antibodies (Quanta Lite aCL IgG, IgM, Inova Diagnostics), and IgG binding to solid-phase and/or phospholipid (PL)-bound beta(2)-GPI and FII by ELISA assays developed an optimized in our laboratory. Odds ratios for the association of IgG binding activity with LA and the aCL IgG status were calculated. In LA patients, dependency of LA potency (as assessed by clotting time prolongation in absence or presence of hexagonal phospholipid) on autoantibody titers was analyzed by the generalized linear model. Total IgG fractions were purified from selected patients to evaluate their ability to inhibit prothrombin activation at low FII concentration. RESULTS: Anticardiolipin antibodies (aCL) of the IgG or IgM type were found in 64 and 23 patients and LA activity in 49 patients. Anti-beta(2)-GPI and anti-FII (solid-phase and PL-bound) IgG titers exceeding by more than 3 standard deviations the mean values observed in control subjects were found in 46 and 47 patients and in 56 and 30 patients respectively, with the highest titers detected in the subgroup of patients with both LA and aCL IgG. The relative risk of LA for patients free of anti-FII and/or anti-beta(2)-GPI IgG was 0.03 after stratification for the aCL IgG status. Anti beta(2)-GPI (solid-phase and PL-bound) IgG (RR 34.4 and 12.6) and anti-FII (solid phase) IgG (RR 6.33) were all associated with LA activity. However, when taking into account co-existence of anti-FII and anti-beta(2)-GPI IgG in the same patients, the relative risk of LA for patients with isolated anti-FII IgG (solid phase and/or PL-bound) was 0.50, whereas it ranged from 4.24 to 8.70 for all the antibody combinations including anti-beta(2)-GPI IgG. Anti-beta(2)-GPI (PL-bound) and aCL IgG titers were the only significant predictors of LA potency determined in absence phospholipid (anti-beta(2)-GPI IgG) or in presence of hexagonal phospholipid (aCL IgG). Total IgG fractions purified from 12 patients (6 with anti-FII IgG) did not significantly inhibit factor II activity up to a 150-fold molar excess. INTERPRETATION AND CONCLUSIONS: These results highlight the high prevalence of anti-FII and anti-beta(2)-GPI IgG in patients with the clinical suspicion of APA syndrome and particularly in the subgroup of patients with LA activity. The fraction of LA activity which can be quenched by addition of hexagonal phospholipid is, however, only dependent on IgG directed to PL-bound beta(2)-GPI. Other antibodies associated with anticardiolipin IgG may explain residual clotting time prolongation observed in the presence of hexagonal phospholipid. PMID- 10477459 TI - Mobilization and selection of peripheral blood hematopoietic progenitors in children with systemic sclerosis. AB - BACKGROUND AND OBJECTIVE: Autologous transplant of lymphocyte-depleted peripheral blood stem cells has been proposed for treatment of patients with severe autoimmune disease. However, until now, no data are available on the safety and feasibility of both stem cell collection and selection in pediatric patients with these disorders. We report on three children affected by systemic sclerosis with lung involvement, who received chemotherapy and granulocyte colony-stimulating factor (G-CSF) to mobilize autologous peripheral blood progenitors. DESIGN AND METHODS: The priming regimen consisted of cyclophosphamide (CY, 4 g/m(2)) and G CSF (lenograstim, 10 microg/kg/day starting 2 days after cyclophosphamide administration until stem cell collection). Leukapheresis was performed when WBC and CD34+ cell count were at least 2 x 10(9)/L and 0.03 x 10(9)/L, respectively. In the first patient, positive selection of CD34+ cells was performed through the Ceprate SC stem cell concentrator (CellPro, Bothell, WA, USA). In the remaining 2 children, progenitor cells were also purged with negative selection of CD4+ and CD8+ lymphocytes performed by means of the Isolex 300i device (Baxter). RESULTS: All patients tolerated the priming regimen well and did not present any sign of autoimmune disease exacerbation. Collection was successful in all children and the number of CD34+ cells before selection ranged between 10.7 x 10(6) and 17.6 x 10(6)/kg of patient body weight. The selection of haematopoietic stem cells in the 3 patients resulted in at least 2. 6-log T-cell depletion of the cell content, with a recovery of the initial value of CD34+ cells comprised between 21 and 44%. After, a preparative regimen consisting of CY (200 mg/kg over 4 days) and Campath-1 G in vivo (10 mg/day for 2 consecutive days), patients were transplanted using cryopreserved lymphocyte-depleted progenitor cells. In all cases, a prompt hematopoietic engraftment was observed. INTERPRETATION AND CONCLUSIONS: Taken together these data suggest that mobilization, collection and selection of hematopoietic progenitors are safe and feasible in children with autoimmune disease. PMID- 10477460 TI - Autologous transplantation in multiple myeloma: a GITMO retrospective analysis on 290 patients. Gruppo Italiano Trapianti di Midollo Osseo. AB - BACKGROUND AND OBJECTIVE: Autologous transplantation is a better treatment for multiple myeloma (MM) than chemotherapy, but uncertainty remains about patient selection, optimal timing of autograft, conditioning regimen, need for a second autograft, and role of maintenance. To provide partial answers to these questions we assessed the results of autologous transplantation in a large cohort of patients whose data were reported to the GITMO registry. DESIGN AND METHODS: We retrospectively analyzed data from 290 patients with MM (M = 150; F = 140; median age 52 years, range 19-70; stage I = 34, stage II = 75, stage III = 167) reported to the GITMO. At the time of autograft, 20% were in CR, 66% in PR, while the remaining had non-responsive or progressive disease. Median time between diagnosis and transplant was 16 months (1-90). Seventy-two patients (26%) had been planned to receive a double autograft, but this was actually done in only 35 (12%). The conditioning was chemotherapy in 90%. Peripheral blood was the only source of stem cells in 94%, and purging was applied in 10% of cases. For statistical analysis of data, differences between patient subsets were analyzed using the chi-square test, while the Kaplan-Meier method was used to estimate event-free survival (EFS) and survival (OS) probabilities. The Cox model was used for multivariate analysis. RESULTS: Following the autograft, 116 patients (40%) were in CR, 144 (50%) in PR, 24 (8%) did not respond or progressed and 6 (2%) died before response evaluation. Transplant-related mortality occurred in 3%. At a median follow-up of 23 months, 223 (77%) patients are alive, 71 (24%) of them in CR, and 67 (23%) patients have died at a median time of 20 months (0-70). OS and EFS at 6 years are 47% and 28%, respectively, but the EFS curve shows no plateau. In multivariate analysis, age, beta2-microglobulin level and status at transplant emerged as significant prognostic factors for both OS and EFS, while time from diagnosis to transplant showed borderline significance. INTERPRETATION AND CONCLUSIONS: Based on the prognostic factors identified in multivariate analysis, we were able to assess the weight of a single prognostic factor or their combinations on transplant outcome. We also calculated the probability of OS and EFS by the number of factors at the time of autograft. Autologous transplantation is a safe and effective procedure, not only in sensitive patients, but also in resistant cases, provided they are <55 years of age and have low beta2-microglobulin. It should be applied early after the diagnosis of multiple myeloma, following the delivery of brief primary chemotherapy. PMID- 10477461 TI - Jugular vein thrombosis: a rare presentation of atypical chronic myeloproliferative disorder in a young woman. AB - Venous thromboembolism is common in subjects with chronic myeloproliferative disorders and is a recognized presenting feature of occult myeloproliferation. We report the case of a young woman who presented with acute thrombosis in the right jugular vein and pulmonary embolism. Splenomegaly and myeloid proliferation with bone marrow fibrosis, in the absence of the criteria for typical myeloproliferative disorders, allowed a diagnosis of an atypical form of chronic myeloproliferative disorder. This form carries a high risk of thrombosis and venous thromboembolism can be the presenting feature, though the course is often indolent. Acute thrombosis in the right jugular vein has not been so far described in these subjects. The outcome of young people with myelofibrosis is unpredictable, but a normal level of hemoglobin and the absence of blast cells and constitutional symptoms at presentation identifies subjects with a low probability of rapid disease progression. PMID- 10477462 TI - Waldenstrom's macroglobulinemia revealed by atypical blood lymphoid cells. PMID- 10477463 TI - Multiple myeloma with Auer-rod-like inclusions. PMID- 10477464 TI - Mansonella perstans filariasis in a HIV patient: finding in bone marrow. PMID- 10477465 TI - Genetic heterogeneity of hereditary stomatocytosis syndromes showing pseudohyperkalemia. PMID- 10477466 TI - In vitro drug-induced cytotoxicity predicts clinical response to high-dose chlorambucil in B-cell chronic lymphocytic leukemia. PMID- 10477467 TI - Extrinsic and intrinsic cholinergic systems of the vascular wall. AB - Immunohistochemical, biochemical and functional studies have revealed two separate cholinergic systems in the arterial vascular wall. Endothelial cells represent the ubiquitous intrinsic, intimal system; they contain the acetylcholine-synthesizing enzyme, choline acetyltransferase, release a choline ester, and contain functional muscarinic receptors. Perivascular autonomic nerve fibres represent the extrinsic, adventitial system. These axons are not ubiquitous but show a highly selective distribution among and even within organs, and utilize co-mediators (NO, neuropeptides) in an organ-specific pattern. We put forward the hypothesis that the intrinsic, intimal system serves as a general regulator of basal vascular tone and wall structure responding to local, luminal stimuli, whereas the perivascular nerve fibres act on top of this basal tone by providing fine tuning in response to reflex activation due to systemic demands. PMID- 10477468 TI - The enteric nervous system: region and target specific projections and neurochemical codes. AB - The goal of this report is to summarise the current knowledge on the projection pathways of enteric neurones innervating the muscle and mucosa in different regions of the gut. Combination of neuronal tracing, immunohistochemical and electrophysiological methods has allowed researchers to gain insight into the enteric hardwiring of specific target tissue in the gut. A polarised innervation pattern of the circular muscle was demonstrated for the stomach fundus/corpus and the ileum with descending pathways being primarily nitrergic while ascending pathways were primarily cholinergic. This characteristic hardwiring is thought to set in part the functional basis for peristalsis. A similar polarised innervation pathway was found for the enteric innervation of the mucosa in the stomach and large intestine but not in the small intestine. In both the stomach (myenteric neurones) and in the proximal and distal colon (submucosal neurones), ascending pathways to the mucosa are primarily cholinergic while descending pathways are primarily non-cholinergic. In the colon, results suggest that activation of both pathways induces a cross potentiation of cholinergic and vasoactive intestinal polypeptidergic mediated secretion. Furthermore, a large population of myenteric neurone s projecting to the mucosa in the small and large intestine are probably intrinsic primary afferent neurones sensitive to mechanical as well as chemical stimuli. PMID- 10477470 TI - Role of Interstitial Cells of Cajal and their relationship with the enteric nervous system. AB - The term 'Interstitial cells of Cajal' (ICC) designates several groups of mesenchymal cells present along the gastro-intestinal tract (GI), in close association with smooth muscle cells and elements of the enteric nervous system (ENS). For years, transmission electron microscopy (TEM) has been the only reliable tool to study ICC. Whilst TEM remains the golden standard for identification of ICC, the observation that the tyrosine kinase receptor c-kit plays a crucial role in their development recently resulted in numerous immunohistochemical studies and also led to a better characterization of their roles. ICC form extensive networks of electrically coupled cells and certain groups of ICC are currently regarded as the source of the spontaneous slow waves of the gut musculature (pacemaker cells). Other ICC appear to be involved in the transduction of the relaxation of smooth muscle triggered by nitric oxide. Abnormal distribution of ICC has been reported in several human diseases and abnormal functioning of ICC might actually be involved in many disorders of GI transit. This review addresses (1) the morphology and relationships of ICC along the GI tract in man and mouse, mainly based on data from immunohistochemistry and confocal microscopy, (2) the emerging role of ICC in the pathophysiology of human diseases, like infantile hypertrophic pyloric stenosis (a common disorder with a dysfunction of the pyloric sphincter), Hirschsprung's disease (aganglion-osis coli) and intestinal pseudo-obstruction, (3) developmental issues, (4) recent reports suggesting a possible link between ICC and gastrointestinal stromal tumors. PMID- 10477469 TI - Species-dependent features of Dogiel type II neurones in the mammalian enteric nervous system. AB - Although autonomic gastrointestinal reflex movements, which occur in all mammalian species, have been described almost a century ago, little was known on the mechanisms underlying this behaviour. Recently, however, intrinsic primary afferent neurones, functioning as the first relay in the reflex arches embedded in the intestinal wall, have been identified in the guinea pig ileum. In guinea pig, such neurones display a Dogiel type II morphology and behave electrophysiologically as slow AHP neurones. In other gastrointestinal regions, in both guinea pig and rat, Dogiel type II cells are also encountered, but the strong correlation with slow AHP neuronal features seems less strict. In large mammals, a correlation of the cellular morphology with intracellular el ectrophysiological recordings has only been obtained in the pig small intestine. Surprisingly, in these experiments aberrant electrophysiological behaviour of Dogiel type II neurones is even more striking since the majority of these cells display electrophysiological features considered typical of S neurones. Furthermore, in those rare cases in which a slow afterhyperpolarization (AHP) could be recorded in porcine Dogiel type II cells, its amplitudes were negligible. This has led us to the conclusion that the differences in electrophysiological behaviour of neurones with comparable morphology in different species are most probably due to the modulating influence of the neurotransmitter substances present. This seems to be the most likely hypothesis in view of the considerable differences in neurotransmitter content of neurones with comparable functions throughout the species. PMID- 10477471 TI - Central autonomic control of the heart, angina, and pathogenic mechanisms of post myocardial infarction depression. AB - Depression can develop in 20% of the patients with a myocardial infarction (MI). Pathobiological mechanisms underlying the development of mood disorders in these patients are unknown. Since post-MI depression has been associated with increased risk of mortality we hypothesized that dysfunction of limbic circuitry is part of the pathogenic processes. Both mood and cardiovascular functions are controlled by the limbic system. Here, we will review a set of experiments that support this hypothesis. Using the retrograde transneuronal transport of pseudorabies virus central autonomic cardiomotor circuitry was identified and Fos protein expression was used for characterization of networks participating in cardiac pain perception, evaluation, and initiation of coping responses. A modified conscious rat model of acute heart pain was employed for induction of cerebral Fos protein expression. Experiments investigating the effects of MI on cerebral activity in the rat showed a selective regional endothelial leakage mainly in the prefrontal cortex, and most severely in the anterior cingulate cortex. This effect was mimicked with intravenous injections of recombinant Tumor Necrosis Factor alpha, which led to the hypothesis that post-MI depression evolves from selective dysfunction of the prefrontal, anterior cingulate cortex in response to (excessive) release of mediators of inflammation. Evidence is provided that cingulate cortex dysfunction may underlie occurrence of mood disorders and derangement of cardiac autonomic control, which would explain the increased risk of mortality associated with post-MI depression. PMID- 10477472 TI - Minority distrust of medicine: a historical perspective. AB - Recent philosophical work has disclosed a host of problems in our apparently natural ways of classifying things. The contemporary classification of certain groups as "minorities" exemplifies some of these problems. I argue that these classifications are arbitrary and misleading. Through examining several of the most significant ethical moments in the history of modern medicine, including the thought and conduct of Nazi physicians, the Tuskegee study, Beecher's questioning of post-war research practices and Percival's enunciation of a universalist ethic for physicians, I make a case against racial and ethnic classification of patients. Such classifications can play a destructive role in determining the sort of health care which minorities receive. Embracing them, even with the intent of improving the lot of those who do not fare well in the present health care environment, is subversive of the egalitarian stance which has been central to medical ethics since Hippocrates. PMID- 10477473 TI - Trusting under pressure. AB - This essay explores the idea that it is possible for a patient to feel ill at ease with a health care professional, even though there is no active ill will on the part of the professional. Noting that the relationship between the patient and the health care professional, especially in the case of the physician, is an asymmetrical one, I suggest that it is incumbent upon professionals to take extra steps to insure that the patient feels at ease in the staff-patient encounter, notwithstanding the good will that health professionals may be assumed to have toward patients generally. PMID- 10477474 TI - Diagnosing death: What's trust got to do with it? AB - Physicians licensed to practice medicine have enjoyed socially endorsed, legally underwritten status-trust to a remarkable degree. However, such trust is not endorsed equally by all segments of American society, most notably, by African Americans. Because physicians underappreciate this fact, they fail to understand how routine medical behavior can disproportionately exacerbate African Americans' pre-existing suspicions. On the other hand, overinterpretation of this fact needlessly risks despair. A theory of trust provides guidance in resolving clinical conflicts. PMID- 10477475 TI - Distrust, social justice, and health care. AB - What steps, if any, should be taken to eliminate the distrust that many racial minorities feel toward the health care system in the United States? Is this distrust an issue of social justice or should it be viewed as an instance where people unreasonably fail to take advantage of existing opportunities? I argue that this distrust is an issue of social justice and that the state does have an obligation to eliminate or mitigate it, especially in the area of public health. PMID- 10477476 TI - Affirmative action and the allocation of health care. AB - The justifications of affirmative action, the compensatory, corrective and redistributive, have been widely recognized in legal thinking. They have been applied, principally, to employment practices. They can also be applied to health care. Arguments for affirmative action in health care allocation shift the burden of proof to those who deny that racism is the root cause of differential morbidity and mortality experienced by, for example, African Americans. At the very least, these arguments mandate much needed research into the causes of minorities' poor health. Without such research, racism remains the presumptive cause of, and affirmative action the appropriate remedy for, the health care problems minorities face. PMID- 10477477 TI - Trust, patient well-being and affirmative action in medical school admissions. AB - This article reviews the current legal arguments for and against affirmative action in medical education. It concludes that many of the traditional legal defenses for race-based admissions are imperiled or defunct. The author suggests that the best and probably most viable justification for affirmative action policies is the one that recognizes that racial diversity in the medical profession is important because it provides the physicians with whom minority patients can feel safe and comfortable. Trust is a central component of the doctor-patient relationship and is the prerequisite, in many cases, to the individual physician's ability to practice good medicine. Unfortunately, minorities' historical and current experience with the medical profession and health delivery system frequently breeds suspicion rather than faith. As a result, society and the medical profession have a compelling interest and duty to produce physicians who can inspire trust in patients. In the short-to-medium term, race-conscious admissions policies may be necessary to fulfill this duty. PMID- 10477478 TI - Asian Patients' distrust of western medical care: one perspective. AB - Asian patients receive, with significant frequency, suboptimal medical care. The sources of shortcomings in the treatment of the Asian American patient are examined in this paper. I argue that it is mainly a failure to interpret patient behavior correctly which causes suboptimal treatment. Such failure stems not from prejudice, but from a lack of understanding of, much less respect for, the systems of thought about health and illness which form the basis for the traditional or tradition-influenced Asian American patient's approach to illness. "Noncompliant" patient behavior is misunderstood if the physician does not grasp the roots of such behavior in a system of beliefs which is not his own. Misunderstanding begets further "noncompliance," initiating a downward spiral. The way out of such spirals lies, I argue, in seeking a more adequate understanding of the patient's beliefs and their behavioral consequences. PMID- 10477479 TI - On establishing trust. AB - Mutual understanding and trust are most easily attained between persons who share a common cultural background. Persons of color often see physicians as members of an alien culture who are not to be trusted. Physicians can and should transcend cultural barriers and secure the trust of patients of color. To do this, a common culture is not required; effort, kindness and sympathy are. PMID- 10477480 TI - The Hispanic, gay, lesbian, bisexual and HIV-infected experience in health care. AB - This presentation, relying both on personal experience and an array of studies, surveys the problems minorities face in trying to obtain adequate health care. From another viewpoint, these are problems that physicians have in trying to provide health care to persons they do not understand and cannot really see or hear. And the problems multiply when the patient is a minority in more than one sense. The gay, Hispanic, HIV+ patient, for example, is removed from the average physician's comprehension to a degree that is itself almost incomprehensible. Treating patients as they ought to be treated requires that physicians overcome many layers of prejudice and unfounded assumptions. Failure to overcome such prejudices distorts medical practice. PMID- 10477481 TI - Patients' rights and professional responsibilities: the moral case for cultural competence. AB - A right to health care can be derived from basic ethical principles. The empirical evidence revealing significant racial inequities in health status, access to health services, quality of care received and outcomes of health services is reviewed. The need for health care providers to acquire cultural competence in order to fulfill their professional responsibilities is discussed; the insight, knowledge and discipline required to function effectively in the context of cultural differences are described. The broader implications of cultural competence for institutional and public policy, research and professional education are outlined. PMID- 10477482 TI - Stop experimenting on my baby! AB - Having a small sick baby in a neonatal intensive care unit can be an extremely difficult experience for any family. A minority family brings to this setting the additional burden of a concern that racism may affect the care their child receives. While the technology may be overwhelming, the unique rules and an apparent disparity in the enforcement of these rules can suggest discrimination. In some cases, these parental perceptions lead to a charge of experimentation. An increased understanding by health care providers of the cultural differences and life experiences that families bring to stressful situations can improve communication. PMID- 10477483 TI - 'Tis better to receive than to give: the relative failure of the African American community to provide organs for transplantation. AB - There is a serious, continuous and increasing shortfall of organs, especially kidneys, for the purpose of transplantation. This shortfall is especially remarkable in African American populations. Because the incidence of hypertension (HTN) and associated end-stage renal diseases (ESRD) is 17 times greater in African Americans, this minority group, which comprises only 12% of the U.S. population, represents 34% of the dialysis population and 30% of the national kidney waiting list. Furthermore, while black individuals comprise 22-24% of kidney recipients, they comprise only 8-11% of donors. Because of the histocompatibility differences between the races and because tissue matching is part of the United Network for Organ Sharing (UNOS) allocation formula, African Americans wait twice as long for kidneys as Caucasians. Also, because they get more poorly matched organs, their kidney transplant graft survival is 10-20% lower than that for other racial groups. The African American community is unaware of the special needs in members of their own race. Steps must be taken to increase minority awareness of the need for well-matched transplant organs and their involvement in the donation process. PMID- 10477484 TI - Blood transfusion and the pregnant Jehovah's witness patient: avoiding a dilemma. AB - The pregnant Jehovah's Witness patient's refusal of lifesaving transfusion creates a conflict for the physician. While legal steps may be initiated to address the problem, a medical approach stressing prophylaxis which anticipates and avoids the ethical dilemma of managing a hemorrhaging pregnant Jehovah's Witness is preferable. PMID- 10477485 TI - African Americans and the medical establishment. AB - The African American community's response to the AIDS epidemic has reflected the profound mistrust of the medical establishment which many African Americans feel. Among African Americans, the belief that the epidemic originated in a genocidal plot is widespread. It is thought that organized medicine has been significantly involved in this plot. If we look at African Americans' historical relationship to the medical establishment from the era of slavery to the recent past, the suspicious attitudes which make such beliefs possible can be seen as an intelligible response to a new disease which disproportionately affects African Americans. Successful medical and public health responses to the epidemic have depended and will continue to depend upon overcoming the historical legacy of suspicion and gaining the trust of the community. PMID- 10477486 TI - Resolving conflicts: misconceptions and myths in the care of the patient with sickle cell disease. AB - Sickle cell disease is an autosomal recessive disease that primarily affects persons of African ancestry. The hallmark of the disease is hemolytic anemia and vaso-occlusive crisis. Patients often have recurrent and severely painful episodes that necessitate the use of opioids. The reluctance of some health care providers to prescribe narcotics has resulted in adversarial relationships with some patients. The socio-cultural disparity between patients and providers may play a role. However, the lack of knowledge and understanding of the underlying pathophysiology of the disease and pain are the key issues. Education, research and hands-on experience, resulting in changes in attitudes and behaviors, will ultimately lead to a more empathic approach to the sickle cell patient. PMID- 10477487 TI - Erythema nodosum. PMID- 10477488 TI - Novel genetic polymorphisms in DNA repair genes: O(6)-methylguanine-DNA methyltransferase (MGMT) and N-methylpurine-DNA glycosylase (MPG) in lung cancer patients from Poland. AB - Individuals with a decreased DNA repair capacity are at increased cancer risk. The aim of our investigation was to detect genetic polymorphisms in DNA repair genes. Two genes, MPG and MGMT, involved in repair of alkylated purines, have been selected. The genetic polymorphisms in the coding exons 2, 3 and 4 of MPG and in the enhancer region of MGMT were searched for in DNA samples from a group of 33 non-small-cell lung cancer (NSCLC) patients from Poland. The PCR products were sequenced with fluorescently labeled terminators and separated on automatic sequencer. Two polymorphisms in MPG were found: in exon 2: CGC-->TGC, (8603C>T, Genbank Accession Z69720) and in exon 3: CCG-->CCA, (12235G>A, Genbank Accession Z69720). The polymorphism in exon 2 results in amino acid substitution (Arg>Cys). Three polymorphisms within or around 59 bp enhancer of MGMT were detected: 1) 1034A>G (Genbank Accession X61657), 2) 1099C>T (Genbank Accession X61657), 3) 79G>T (Genbank Accession U95038). Polymorphism 2 is located in the 59-bp enhancer sequence, within a palindrome GGTGCGCACC. Polymorphism 3 destroys an inverted repeat GGGTGGGGGGCCGCCCTGACCCCCACCC that contains two PuF binding sequences GGGTGGG separated by Sp1 site. The nature and location of these polymorphisms is consistent with the hypothesis that they may have functional significance. PMID- 10477489 TI - A novel frameshift mutation (1651ins5) in exon 10 of the CFTR gene can be misinterpreted as a DeltaF508 mutation. PMID- 10477490 TI - A novel missense mutation Ile193Val in cholesteryl ester transfer protein gene. PMID- 10477491 TI - A novel missense mutation Thr316Ala in lysosomal acid lipase gene in Japanese population. PMID- 10477492 TI - Identification of the new polymorphism IVS1-91 C-->T in the beta globin gene. PMID- 10477493 TI - Mutation profile in the beta-myosin heavy chain gene in hypertensive hypertrophic heart disease. PMID- 10477494 TI - Run-on mutation and three novel nonsense mutations identified in the PAX6 gene in patients with aniridia. PMID- 10477495 TI - Consensus statement: nutritional morbidity in children with cancer. PMID- 10477496 TI - Issues of health care mistrust in East Harlem. AB - Barriers to the delivery of health care services to the Latino community are described and analyzed. Fear of deportation, unfamiliarity with modern medicine, inability to speak English and reliance upon traditional curing practices all contribute to the harmful underutilization of medical services which we see in the Latino community. Health care providers who serve that community need to overcome these obstacles by expanding their cultural awareness and increasing the effectiveness of their communication. PMID- 10477497 TI - Medicine and mobility: challenge for the next century. AB - Advances in air traffic are the key factors that have enabled society during the present century to change from local orientation to globalization. At the end of the 20th century, millions can individually travel across continents into different cultures, climates, environments, and reach their destination within hours or, if it is real remote, in a few days. Advances in telecommunication and informatics technologies have furthermore allowed us to be in contact in real time wherever on, below or above earth s surface humans are. These progresses influence all aspects of human life. They also bring numerous chances, challenges and problems for medicine. Therefore, the different aspects of increased mobility should be viewed in a concerted fashion, that integrates all aspects of medicine that are influenced by the increased mobility of society. This new field of medicine, "Medicine and Mobility", will have a central role to fulfill the main task of medicine during the next century, namely to make individualized medicine possible, i.e. to bring individualized medical expertise to the patient, wherever he/she may be. This will end the present expertise-oriented medicine of the 20th Century, in which the patient has to come to the special expertise. This change of medical practice will have huge impacts not only on medicine, but on economies, cultures and be one of the driving forces for our main task of the next century, to peacefully unite our whole globe. PMID- 10477498 TI - Preparing for Mars: the physiologic and medical challenges. AB - As the twentieth century closes, retrospectives cite the Apollo moon missions as one of the important events of the past 100 years. A trip to Mars, however, would be even more challenging and significant. A round-trip Mars journey would require nearly three years away from Earth, a significant leap in complexity compared to the two week long Moon trips or the record-breaking fourteen-month flight on Mir. What would be the physiologic and medical challenges of a Mars flight? Two key areas of physiology present the greatest potential problems--calcium metabolism and radiation exposure. Data from Mir missions show that bone loss continues in space despite an aggressive countermeasure program. Average losses were 0.35% per month, but some load bearing areas lost >1% per month. A 1% loss rate, if it continued unabated for 30 months, could produce osteoporosis. Smaller losses could still increase fracture risk. Some bone loss can be well tolerated, particularly if the bone can be regained after the mission. But the effectiveness of post-flight rehabilitation to restore the density and quality of bone after spaceflight is not well known. Bone loss estimates are based on continuous weightlessness exposure, but this is not a requirement for a Mars trip. Most of the time on a Mars trip will be spent in the 1/3 Earth's gravity environment on Mars, and either intermittent or continuous artificial gravity can be provided for the transit between planets (although at an engineering cost). The dosing of the gravity exposure (e.g. the level and duration), however, has not been established. Radiation protection also requires a balance between engineering cost and human health. Excessive shielding could add billions of dollars to the cost of a mission. Trips in interplanetary space, however, expose the crew to heavy high-energy particles from cosmic rays (HZE particles), which have a high linear energy transfer. This high energy leads to significant biological damage (e.g. chromosomal aberrations, cancer induction). A recent report from the Committee on Space Biology and Medicine notes that only one systematic study of cancer induction from high-energy particles has been conducted (using the mouse Harderian gland). Predictions of cancer risk and acceptable radiation exposure in space are extrapolated from minimal data. Other areas of physiology also present problems, such as muscle loss, cardiovascular deconditioning, and vestibular adaptation. Despite all the issues, however, a focussed, aggressive research program that uses the resources of the International Space Station should pave the way for mankind's greatest adventure--a trip to Mars. PMID- 10477499 TI - Calcium metabolism in microgravity. AB - Unloading of weight bearing bones as induced by microgravity or immobilization has significant impacts on the calcium and bone metabolism and is the most likely cause for space osteoporosis. During a 4.5 to 6 month stay in space most of the astronauts develop a reduction in bone mineral density in spine, femoral neck, trochanter, and pelvis of 1%-1.6% measured by Dual Energy X-ray Absorption (DEXA). Dependent on the mission length and the individual turnover rates of the astronauts it can even reach individual losses of up to 14% in the femoral neck. Osteoporosis itself is defined as the deterioration of bone tissue leading to enhanced bone fragility and to a consequent increase in fracture risk. Thinking of long-term missions to Mars or interplanetary missions for years, space osteoporosis is one of the major concerns for manned spaceflight. However, decrease in bone density can be initiated differently. It either can be caused by increases in bone formation and bone resorption resulting in a net bone loss, as obtained in fast looser postmenopausal osteoporosis. On the other hand decrease in bone formation and increase in bone resorption also leads to bone losses as obtained in slow looser postmenopausal osteoporosis or in Anorexia Nervosa patients. Biomarkers of bone turnover measured during several missions indicated that the pattern of space osteoporosis is very similar to the pattern of Anorexia Nervosa patients or slow looser postmenopausal osteoporosis. However, beside unloading, other risk factors for space osteoporosis exist such as stress, nutrition, fluid shifts, dehydration and bone perfusion. Especially nutritional factors may contribute considerably to the development of osteoporosis. From earthbound studies it is known that calcium supplementation in women and men can prevent bone loss of 1% bone per year. Based on these results we studied the calcium intake during several European missions and performed an experiment during the German MIR 97 mission where we investigated the effects of high calcium intake (>1000 mg/d) and vitamin D supplementation (650 IU/d) on the calcium and bone metabolism during 21 days in microgravity. In the MIR 97 mission high calcium intake and vitamin D supplementation led to high ionized calcium levels and a marked decrease in calcitriol levels together with decreased bone formation and increased bone resorption markers. Our conclusion from the MIR 97 mission is that an adequate calcium intake and vitamin D supplementation during space missions is mandatory but, in contrast to terrestrial conditions, does not efficiently counteract the development of space osteoporosis. PMID- 10477501 TI - Research on humans during space flights: ethical problems. AB - Experiments on humans during space flights call for ethical reflection, which is more than a mere assessment of the risks and a concurrent ethical evaluation of manned space flight as such. Although it may be desirable to harmonize ethical standards for international space flights especially against the background of the construction of an International Space Station (ISS) the application of ethical standards for human experiments on earth to the situation in space is not (at all) straight forward. The analysis shows that an ethical evaluation of experiments in space involving humans cannot be made solely on grounds of those principles which apply in comparable cases of terrestrial experiments. The very special conditions in space necessitate further ethical considerations. From this it may of course not be inferred that the current practice of strict ethical evaluation of experiments on humans becomes obsolete once we leave the planet earth. On the contrary, it means that widely accepted principles--like free and informed consent, or data protection and privacy--may need to be amended in order to take proper account of the special conditions in space and the need to achieve a consensus among the different persons involved. PMID- 10477500 TI - Human immune cells as space travelers. AB - Experiments in space have shown that T lymphocyte function is altered in more than 50% of space crew members. There is strong evidence that such effect is due to stress rather than to weightlessness per se. However the health of astronauts was never threatened so far. Experiments in-vitro with cultures of human peripheral blood lymphocytes (not from astronauts) have shown that T cell function is dramatically reduced. Recent work with the random positioning machine, a new instrument to simulate conditions similar to microgravity, indicate that there are direct gravitational effects on the genetic expression of interleukin-2 and of its receptor in T lymphocytes. PMID- 10477502 TI - A study on the effect of oxygen bolus breathing or pressure breathing on oxygen saturation, at rest or under physical strain (bicycle ergometer) at 14,000 ft altitude. AB - At an altitude of about 3000 m even healthy people suffer from hypoxemia due to the low oxygen partial pressure. As a physiological countermeasure the oxygen partial pressure is increased. In the present study the following procedures are looked into: 1. Pressurized breathing, 2. Oxygen bulus breathing. It was found out that both procedures are appropriate to increase the oxygen partial pressure efficiently. However, bolus breathing is more effective, since an almost 100% hemoglobin saturation may be achieved even with small bolus volumes when the subject is at rest or under little physical strain. Pressurized breathing seems to be an appropriate emergency measure when a sudden pressure loss occurs in an aircraft for a short while provided that the subject is familiar with the breathing technique. PMID- 10477503 TI - Lung function requirements in flying duty the problem of bronchial hyperresponsiveness in military aircrew. AB - Uncompromised lung function is essential for fitness to fly. Under hypobaric conditions there is an increased risk of hypoxemia. G-forces, positive pressure breathing and anti-G maneuvers cause physical stress to the lung tissue and altered pulmonary blood flow. Breathing with pure oxygen, dry cabin air and ozone can cause airway irritation. Chemically and physically by irritating agents may be present. Emergencies such as smoke in the cockpit or inhalation of tear gas can rapidly compromise the pulmonary system in susceptible persons. Sudden incapacitation may occur. Trapped gases may cause overinflation and lung rupture in rapid decompression. Applicants for military duty have to pass basic lung function tests routinely. Preselection of aircrew candidates tends to be even stricter. Asthma and obstructive lung disease are disqualifying. Trained aircrew with late onset of pulmonary problems can be waived under certain restrictions in many cases. Some national regulations exclude even applicants with allergies. Due to aeromedical experience we should always be aware of the latent unspecific bronchial hyperresponsiveness (BHR). BHR is one of the characteristics of asthma bronchiale. If BHR exists there is an increased risk of later development of asthma bronchiale, especially together with perennial allergies such as against house dust mite. Under certain conditions BHR can become symptomatic and aeromedically relevant. In some cases we saw an exacerbation under medication, mostly under beta-receptor-blockers. In one case even under betablocker containing eye drops. In the Gulf War 1991 a number of allied military personnel had to be withdrawn because of bronchospastic symptoms. This can be explained among others by medication with physostigmine. Physostigmine is a systemically active cholinergic drug which is prophylactically used under threat of chemically warfare agents. In individuals with latent BHR physostigmine will lower the threshold for bronchial reactions considerably and even cause manifest bronchospasm. CONCLUSIONS: We recommend an unspecific bronchial challenge test in the selection of personnel for duties where uncompromised lung function under all environmental conditions is essential. PMID- 10477504 TI - Effects of an antihypertensive medication on functional capacity under simulated flight-typical stress-conditions. AB - A model to investigate the functional capacity (psychomental performance) under stressful conditions was developed. Twenty eight patients with mild hypertension receiving Nitrendipine (20 mg) for 30 days were tested under hypoxic (16% oxygen) and/or orthostatic (-30 mmHg lower body negative pressure) conditions using a subset of the AGARD battery. The main effect was a decreasing performance of the grammatical reasoning task (GRT) under hypoxia or the combination of hypoxia and orthostasis. A simultaneous application of stressors while performing psychometric test batteries may be useful to reveal pharmaceutical influences on human performance and may help to recommend the use of drugs in occupational medicine. PMID- 10477505 TI - New refractive surgery procedures in ophthalmology and the influence on Pilot's fitness for flying. AB - During the last years more and more procedures came up to render people with refractive errors possible to see without contact lenses or glasses. The different procedures for hyperopia, myopia and astigmatism are performed in increasing numbers in hospitals, laser-centres and in private practise. Modern radial keratotomy was introduced in the late 70 by the Russian Fjodorov. Because of the many complications this procedure was replaced by other procedures. Photorefractive keratectomy (PRK) was introduced in 1985, using an excimer laser to vaporize corneal tissue. Because of the side effects as pain and glare and the limited indication area Laser in situ keratomileusis (LASIK) was developed. In this procedure a corneal lamella is cut in, flapped back and the excimer laser vaporize the corneal tissue. These two procedures are mostly applied nowadays. But there are also new techniques that are still in an experimental state. One of that is the implementation of an intrastromal corneal ring that can reduce myopia up to 4 diopters. During the laser thermo keratoplasty 8 to 16 laser applications are performed in the periphery of the cornea to reduce hyperopia up to + 5 diopters. Another refractive surgery procedure is the phakic intraocular lens implantation for high myopia and +3 to +10 diopters hyperopia and the clear lens extraction with intraocular lens implantation for high hyperopia. The Joint Aviation Authority Requirements allow a hyperopia and myopia for commercial pilots of 3 diopters. Some pilot candidates with higher refractive errors undergo refractive surgery. But there are also pilots who are customers of the refractive operating ophthalmologists to get rid of the crutch glasses. This paper gives an overview about the refractive procedures that are performed nowadays on pilots and pilot candidates and gives information about the relevant indications and complications of refractive procedures in the aviation environment. PMID- 10477506 TI - Melatonin and jet-lag. AB - Caused by time shift, a desynchronisation of the body clock from external zeitgebers occurs after transmeridian flight which leads to disturbances of sleep and circadian rhythms. These disturbances are not pathological and diminish within days. To achieve a faster resynchronisation than naturally, the hormone melatonin is often taken by business people and travelers, and--to some extent- by aircrew. The usefulness of the melatonin intake for alleviation of jet-lag is intensively discussed. Most field studies reporting about a favourable influence of melatonin on jet-lag, have been performed using questionnaires; few studies monitoring physiological circadian functions have found a better adjustment under melatonin treatment. However, from laboratory experiments is known that external melatonin is indeed capable to influence the circadian system. With respect to the efficacy of melatonin on better sleep and performance, there is a lack of information from field studies, and laboratory studies do not provide consistent results. Unequivocal estimations of the dosage of melatonin for best efficacy are not yet performed, although a range of different dosages have been tested. Recommendations about dosage, duration of medication and time of intake (which is of major importance for efficacy) do not rely on systematic examinations of the drug. Adverse effects of melatonin on sleepiness and impaired performance directly after intake of the drug are known. From studies is also derived that an inappropriate timing of intake causes sleep disturbances and unfavourable shifts of the circadian system. The administration of melatonin for influencing sleep and circadian rhythms cannot be recommended for aircrew. Flight physicians should refer to adverse and side effects of the hormone melatonin. Before any general recommendations for the use of melatonin can be presented, genuine clinical studies following good clinical practice should be performed. PMID- 10477507 TI - Origin of back pain during bedrest: A new hypothesis. AB - Back pain during bed rest has frequently been reported. Until recently, studies concentrated mainly on length changes of the spine. Functional aspects such as reduced movements of the back, although anecdotally reported, could neither be quantified nor were they taken into account as a potential cause of back pain. In the present study the geometry of the spinal columns of eight healthy men was continuously recorded over 24 h before, during, and after a seven-day 6 degree head-down tilt period (Series 1). The approach is based on the measurement of skin distances between four pairs of miniaturized ultrasound transmitters and receivers fixed on the back in parallel to the thoracic and lumbar spine. After a four-week break, the entire study was repeated with the same subjects (Series 2). The major findings are: 1. The length increase of the spine segments investigated was virtually identical in both series. 2. During the bedrest period significantly greater pain occurred in Series 1 as compared to Series 2. Pain was generally confined to the lumbar region about 10 cm lateral of the spinous processes. 3. Pain was most prominent when trunk motions were minimal, while movements over the maximal range of the spine transiently reduced the pain remarkably. 4. During bed rest, the amplitude of voluntary movements of the lumbar segment was reduced in particular during Series 1. It is concluded that back pain during bed rest is induced by reduced amplitudes of spine movements rather than by increases in the spine length. It is suggested that isometric muscle contractions of low intensity are a primary course of back pain. Regular, slow, large-amplitude movements of the spine are recommended as a countermeasure. PMID- 10477508 TI - Telemedical support of prehospital emergency care in mass casualty incidents. AB - In the German emergency medical service system (EMSS) medical treatment can be improved in most of mass casualty incidents (MCI). Currently, the incident commander who is responsible for classification of the victims (depending on their urgency and condition, the so called triage) and ordered transportation uses paper-based documentation. Triage tags are used to identify and classify patients and gather treatment information. This can cause problems in medical treatment and in transportation of injured victims. Object-oriented modelling, simulation, and visualisation of processes can show deficits in treatment and data processing and thereby help to optimise medical workflow and logistics. If documentation by paramedics and emergency physicians is done electronically, all patient records could be send to a telemedical centre for central data administration. A telemedical supported triage tag helps identifying victims and managing detailed identification protocols. The paper-based documentation in emergency would become obsolete, if hospitals can query all protocols, diagnoses, and findings from the telemedical centre. Safety and security aspects can be guaranteed. The complete medical treatment workflow can be supported by telemedicine. Therefore, in case of MCI, telemedicine can optimise medical treatment and exonerate the paramedics from unnecessary documentation. PMID- 10477509 TI - Travel medicine--the next 10 years. AB - In summary, the development of travel medicine in the next years will be characterized by new risks, and on the other hand by new methods of therapy and prophylaxis. Formally, travel medicine will be established as an interdisciplinary special discipline, while the know-how in travel medicine will be consolidated by certificates. PMID- 10477510 TI - Postdoctoral patterns, career advancement, and problems. AB - Postdoctoral appointments can have different functions and meanings, depending on the field and whether the postdoc is a man or a woman. The Ph.D.'s-Ten Years Later study confirmed that in biochemistry, the postdoc, not the Ph.D., has become the general proving ground for excellence both in academia and industry. Because they spent a longer time in these "mandatory" postdocs, biochemists had the largest proportion of untenured faculty 10 to 13 years after the Ph. D. In mathematics, where substantially fewer postdoctoral positions are available, Ph.D.'s taking postdocs are more likely to obtain faculty positions, but this is true only for men. University administrators should be accountable for monitoring the total time spent in these positions and should provide administrative assistance for skills training, career growth, and the job search. In addition, creative solutions concerning the dual-career couple phenomenon are necessary. PMID- 10477511 TI - Single-Mode Photonic Band Gap Guidance of Light in Air. AB - The confinement of light within a hollow core (a large air hole) in a silica-air photonic crystal fiber is demonstrated. Only certain wavelength bands are confined and guided down the fiber, each band corresponding to the presence of a full two-dimensional band gap in the photonic crystal cladding. Single-mode vacuum waveguides have a multitude of potential applications from ultrahigh-power transmission to the guiding of cold atoms. PMID- 10477512 TI - Spatially Inhomogeneous Metal-Insulator Transition in Doped Manganites. AB - Scanning tunneling spectroscopy was used to investigate single crystals and thin films of La(1-x)Ca(x)MnO(3) (with x of about 0.3), which exhibit colossal magnetoresistance. The different spectroscopic signatures of the insulating (paramagnetic) and metallic (ferromagnetic) phases enable their spatial extent to be imaged down to a lateral scale of the order of 10 nanometers. Above the bulk transition temperature T(c), the images show mostly insulating behavior. Below T(c), a phase separation is observed where inhomogeneous structures of metallic and more insulating areas coexist and are strongly field dependent in their size and structure. Insulating areas are found to persist far below T(c). These results suggest that the transition and the associated magnetoresistance behavior should be viewed as a percolation of metallic ferromagnetic domains. PMID- 10477513 TI - Stable five- and six-coordinated silicate anions in aqueous solution. AB - Addition of aliphatic polyols to aqueous silicate solutions is shown to yield high concentrations of stable polyolate complexes containing five- or six coordinated silicon. Coordinating polyols require at least four hydroxy groups, two of which must be in threo configuration, and coordinate to silicon via hydroxy oxygens at chain positions on either side of the threo pair. The remarkable ease by which these simple sugar-like molecules react to form hypervalent silicon complexes in aqueous solution supports a long-standing supposition that such species play a significant role in the biological uptake and transport of silicon and in mineral diagenesis. PMID- 10477514 TI - Seismic Velocity and Density Jumps Across the 410- and 660-Kilometer Discontinuities. AB - The average seismic velocity and density jumps across the 410- and 660-kilometer discontinuities in the upper mantle were determined by modeling the observed range dependence in long-period seismic wave arrivals that reflect off of these interfaces. The preliminary reference Earth model (PREM) is within the computed 95 percent confidence ellipse for the 410-km discontinuity but outside the allowed jumps across the 660-kilometer discontinuity. Current pyrolite mantle models appear consistent with the constraints for the 410-kilometer discontinuity but overpredict amplitudes for the 660-kilometer reflections. The density jump across the 660-kilometer discontinuity is between 4 and 6 percent, below the PREM value of 9.3 percent commonly used in mantle convection calculations. PMID- 10477515 TI - Improved Weather and Seasonal Climate Forecasts from Multimodel Superensemble. AB - A method for improving weather and climate forecast skill has been developed. It is called a superensemble, and it arose from a study of the statistical properties of a low-order spectral model. Multiple regression was used to determine coefficients from multimodel forecasts and observations. The coefficients were then used in the superensemble technique. The superensemble was shown to outperform all model forecasts for multiseasonal, medium-range weather and hurricane forecasts. In addition, the superensemble was shown to have higher skill than forecasts based solely on ensemble averaging. PMID- 10477516 TI - Formation of Ordered Nanoscale Semiconductor Dots by Ion Sputtering. AB - A formation process for semiconductor quantum dots based on a surface instability induced by ion sputtering under normal incidence is presented. Crystalline dots 35 nanometers in diameter and arranged in a regular hexagonal lattice were produced on gallium antimonide surfaces. The formation mechanism relies on a natural self-organization mechanism that occurs during the erosion of surfaces, which is based on the interplay between roughening induced by ion sputtering and smoothing due to surface diffusion. PMID- 10477517 TI - KDR receptor: a key marker defining hematopoietic stem cells. AB - Studies on pluripotent hematopoietic stem cells (HSCs) have been hindered by lack of a positive marker, comparable to the CD34 marker of hematopoietic progenitor cells (HPCs). In human postnatal hematopoietic tissues, 0.1 to 0.5% of CD34(+) cells expressed vascular endothelial growth factor receptor 2 (VEGFR2, also known as KDR). Pluripotent HSCs were restricted to the CD34+KDR+ cell fraction. Conversely, lineage-committed HPCs were in the CD34+KDR- subset. On the basis of limiting dilution analysis, the HSC frequency in the CD34+KDR+ fraction was 20 percent in bone marrow (BM) by mouse xenograft assay and 25 to 42 percent in BM, peripheral blood, and cord blood by 12-week long-term culture (LTC) assay. The latter values rose to 53 to 63 percent in LTC supplemented with VEGF and to greater than 95 percent for the cell subfraction resistant to growth factor starvation. Thus, KDR is a positive functional marker defining stem cells and distinguishing them from progenitors. PMID- 10477519 TI - Historical Genetics: The Parentage of Chardonnay, Gamay, and Other Wine Grapes of Northeastern France. AB - The origins of the classic European wine grapes (Vitis vinifera) have been the subject of much speculation. In a search for parental relationships, microsatellite loci were analyzed in more than 300 grape cultivars. Sixteen wine grapes that have long been grown in northeastern France, including 'Chardonnay', 'Gamay noir', 'Aligote', and 'Melon', have microsatellite genotypes consistent with their being the progeny of a single pair of parents, 'Pinot' and 'Gouais blanc', both of which were widespread in this region in the Middle Ages. Parentage analysis at 32 microsatellite loci provides statistical support for these relationships. PMID- 10477518 TI - Whole-genome shotgun optical mapping of Deinococcus radiodurans. AB - A whole-genome restriction map of Deinococcus radiodurans, a radiation-resistant bacterium able to survive up to 15,000 grays of ionizing radiation, was constructed without using DNA libraries, the polymerase chain reaction, or electrophoresis. Very large, randomly sheared, genomic DNA fragments were used to construct maps from individual DNA molecules that were assembled into two circular overlapping maps (2.6 and 0.415 megabases), without gaps. A third smaller chromosome (176 kilobases) was identified and characterized. Aberrant nonlinear DNA structures that may define chromosome structure and organization, as well as intermediates in DNA repair, were directly visualized by optical mapping techniques after gamma irradiation. PMID- 10477520 TI - Differential stimulation of PKC phosphorylation of potassium channels by ZIP1 and ZIP2. AB - Targeting of protein modification enzymes is a key biochemical step to achieve specific and effective posttranslational modifications. Two alternatively spliced ZIP1 and ZIP2 proteins are described, which bind to both Kvbeta2 subunits of potassium channel and protein kinase C (PKC) zeta, thereby acting as a physical link in the assembly of PKCzeta-ZIP-potassium channel complexes. ZIP1 and ZIP2 differentially stimulate phosphorylation of Kvbeta2 by PKCzeta. They also interact to form heteromultimers, which allows for a hybrid stimulatory activity to PKCzeta. Finally, ZIP1 and ZIP2 coexist in the same cell type and are elevated differentially by neurotrophic factors. These results provide a mechanism for specificity and regulation of PKCzeta-targeted phosphorylation. PMID- 10477521 TI - In vivo protein transduction: delivery of a biologically active protein into the mouse. AB - Delivery of therapeutic proteins into tissues and across the blood-brain barrier is severely limited by the size and biochemical properties of the proteins. Here it is shown that intraperitoneal injection of the 120-kilodalton beta galactosidase protein, fused to the protein transduction domain from the human immunodeficiency virus TAT protein, results in delivery of the biologically active fusion protein to all tissues in mice, including the brain. These results open new possibilities for direct delivery of proteins into patients in the context of protein therapy, as well as for epigenetic experimentation with model organisms. PMID- 10477522 TI - Inhibitors of the nonmevalonate pathway of isoprenoid biosynthesis as antimalarial drugs. AB - A mevalonate-independent pathway of isoprenoid biosynthesis present in Plasmodium falciparum was shown to represent an effective target for chemotherapy of malaria. This pathway includes 1-deoxy-D-xylulose 5-phosphate (DOXP) as a key metabolite. The presence of two genes encoding the enzymes DOXP synthase and DOXP reductoisomerase suggests that isoprenoid biosynthesis in P. falciparum depends on the DOXP pathway. This pathway is probably located in the apicoplast. The recombinant P. falciparum DOXP reductoisomerase was inhibited by fosmidomycin and its derivative, FR-900098. Both drugs suppressed the in vitro growth of multidrug resistant P. falciparum strains. After therapy with these drugs, mice infected with the rodent malaria parasite P. vinckei were cured. PMID- 10477523 TI - Functional interaction of BRCA1-associated BARD1 with polyadenylation factor CstF 50. AB - Polyadenylation of messenger RNA precursors requires a complex protein machinery that is closely integrated with the even more complex transcriptional apparatus. Here a polyadenylation factor, CstF-50 (cleavage stimulation factor), is shown to interact in vitro and in intact cells with a nuclear protein of previously unknown function, BRCA1-associated RING domain protein (BARD1). The BARD1-CstF-50 interaction inhibits polyadenylation in vitro. BARD1, like CstF-50, also interacts with RNA polymerase II. These results indicate that BARD1-mediated inhibition of polyadenylation may prevent inappropriate RNA processing during transcription, perhaps at sites of DNA repair, and they reveal an unanticipated integration of diverse nuclear events. PMID- 10477524 TI - Control of circadian rhythms and photoperiodic flowering by the Arabidopsis GIGANTEA gene. AB - Photoperiodic responses in plants include flowering that is day-length-dependent. Mutations in the Arabidopsis thaliana GIGANTEA (GI) gene cause photoperiod insensitive flowering and alteration of circadian rhythms. The GI gene encodes a protein containing six putative transmembrane domains. Circadian expression patterns of the GI gene and the clock-associated genes, LHY and CCA1, are altered in gi mutants, showing that GI is required for maintaining circadian amplitude and appropriate period length of these genes. The gi-1 mutation also affects light signaling to the clock, which suggests that GI participates in a feedback loop of the plant circadian system. PMID- 10477525 TI - Real-time tracking of memory formation in the human rhinal cortex and hippocampus. AB - A fundamental question about human memory is which brain structures are involved, and when, in transforming experiences into memories. This experiment sought to identify neural correlates of memory formation with the use of intracerebral electrodes implanted in the brains of patients with temporal lobe epilepsy. Event related potentials (ERPs) were recorded directly from the medial temporal lobe (MTL) as the patients studied single words. ERPs elicited by words subsequently recalled in a memory test were contrasted with ERPs elicited by unrecalled words. Memory formation was associated with distinct but interrelated ERP differences within the rhinal cortex and the hippocampus, which arose after about 300 and 500 milliseconds, respectively. These findings suggest that declarative memory formation is dissociable into subprocesses and sequentially organized within the MTL. PMID- 10477526 TI - Role of angiotensin receptor blockers in heart failure: not yet RESOLVD. PMID- 10477527 TI - Incremental prognostic value of post-stress left ventricular ejection fraction and volume by gated myocardial perfusion single photon emission computed tomography. AB - BACKGROUND: The incremental prognostic value of post-stress left ventricular ejection fraction (EF) and volume over perfusion has not been investigated. METHODS AND RESULTS: We identified 1680 consecutive patients who underwent rest Tl-201/stress Tc-99m sestamibi gated single photon emission computed tomography (SPECT) and who were followed-up for 569+/-106 days. Receiver-operator characteristics analysis defined an EF<45%, an end-systolic volume (ESV) >70 mL, and an end-diastolic volume >120 mL as optimal thresholds, yielding moderate sensitivity and high specificity in the prediction of cardiac death. Patients with an EF> or = 45% had mortality rates <1%/year, despite severe perfusion abnormalities, whereas patients with an EF<45% had high mortality rates, even with only mild/moderate perfusion abnormalities (9.2%/year; P<0.00001). Similarly, an ESV< or = 70 mL was related to a low cardiac death rate (<1.2%/year), even for patients with severe perfusion abnormalities, whereas patients with an ESV>70 mL and only mild/moderate perfusion abnormalities had high death rates (8.2%/year; P<0.00001). Patients with an EF<45% and an ESV< or = 70 mL had low cardiac death rates (1.7%/year); those with an EF<45% but an ESV>70 mL had high death rates (7.9%/year; P<0.02). Multivariate Cox proportional hazards regression showed that perfusion variables and ESV were independent predictors of overall coronary events, whereas EF and ESV demonstrated incremental prognostic values over prescan and perfusion information in predicting cardiac death and cardiac death or myocardial infarction. CONCLUSIONS: Post-stress EF and ESV by gated-SPECT have incremental prognostic values over prescan and perfusion information in predicting cardiac death, and they provide clinically useful risk stratification. PMID- 10477528 TI - Derivation and prospective validation of a simple index for prediction of cardiac risk of major noncardiac surgery. AB - BACKGROUND: Cardiac complications are important causes of morbidity after noncardiac surgery. The purpose of this prospective cohort study was to develop and validate an index for risk of cardiac complications. METHODS AND RESULTS: We studied 4315 patients aged > or = 50 years undergoing elective major noncardiac procedures in a tertiary-care teaching hospital. The main outcome measures were major cardiac complications. Major cardiac complications occurred in 56 (2%) of 2893 patients assigned to the derivation cohort. Six independent predictors of complications were identified and included in a Revised Cardiac Risk Index: high risk type of surgery, history of ischemic heart disease, history of congestive heart failure, history of cerebrovascular disease, preoperative treatment with insulin, and preoperative serum creatinine >2.0 mg/dL. Rates of major cardiac complication with 0, 1, 2, or > or = 3 of these factors were 0.5%, 1.3%, 4%, and 9%, respectively, in the derivation cohort and 0.4%, 0.9%, 7%, and 11%, respectively, among 1422 patients in the validation cohort. Receiver operating characteristic curve analysis in the validation cohort indicated that the diagnostic performance of the Revised Cardiac Risk Index was superior to other published risk-prediction indexes. CONCLUSIONS: In stable patients undergoing nonurgent major noncardiac surgery, this index can identify patients at higher risk for complications. This index may be useful for identification of candidates for further risk stratification with noninvasive technologies or other management strategies, as well as low-risk patients in whom additional evaluation is unlikely to be helpful. PMID- 10477529 TI - Purple grape juice improves endothelial function and reduces the susceptibility of LDL cholesterol to oxidation in patients with coronary artery disease. AB - BACKGROUND: In vitro, the flavonoid components of red wine and purple grape juice are powerful antioxidants that induce endothelium-dependent vasodilation of vascular rings derived from rat aortas and human coronary arteries. Although improved endothelial function and inhibition of LDL oxidation may be potential mechanisms by which red wine and flavonoids reduce cardiovascular risk, the in vivo effects of grape products on endothelial function and LDL oxidation have not been investigated. This study assessed the effects of ingesting purple grape juice on endothelial function and LDL susceptibility to oxidation in patients with coronary artery disease (CAD). METHODS AND RESULTS: Fifteen adults with angiographically documented CAD ingested 7.7+/-1.2 mL. kg(-1). d(-1) of purple grape juice for 14 days. Flow-mediated vasodilation (FMD) was measured using high resolution brachial artery ultrasonography. Susceptibility of LDL particles to oxidation was determined from the rate of conjugated diene formation after exposure to copper chloride. At baseline, FMD was impaired (2.2+/-2. 9%). After ingestion of grape juice, FMD increased to 6.4+/-4.7% (P=0.003). In a linear regression model that included age, artery diameter, lipid values, and use of lipid-lowering and antioxidant therapies, the effect of grape juice on FMD remained significant (mean change 4.2+/-4.4%, P<0.001). After ingestion of grape juice, lag time increased by 34.5% (P=0.015). CONCLUSIONS: Short-term ingestion of purple grape juice improves FMD and reduces LDL susceptibility to oxidation in CAD patients. Improved endothelium-dependent vasodilation and prevention of LDL oxidation are potential mechanisms by which flavonoids in purple grape products may prevent cardiovascular events, independent of alcohol content. PMID- 10477530 TI - Comparison of candesartan, enalapril, and their combination in congestive heart failure: randomized evaluation of strategies for left ventricular dysfunction (RESOLVD) pilot study. The RESOLVD Pilot Study Investigators. AB - BACKGROUND: We investigated the effects of candesartan (an angiotensin II antagonist) alone, enalapril alone, and their combination on exercise tolerance, ventricular function, quality of life (QOL), neurohormone levels, and tolerability in congestive heart failure (CHF). METHODS AND RESULTS: Seven hundred sixty-eight patients in New York Heart Association functional class (NYHA FC) II to IV with ejection fraction (EF) <0.40 and a 6-minute walk distance (6MWD) <500 m received either candesartan (4, 8, or 16 mg), candesartan (4 or 8 mg) plus 20 mg of enalapril, or 20 mg of enalapril for 43 weeks. There were no differences among groups with regard to 6MWD, NYHA-FC, or QOL. EF increased (P=NS) more with candesartan-plus-enalapril therapy (0.025+/-0.004) than with candesartan alone (0.015+/-0.004) or enalapril alone(0.015+/-0.005). End diastolic (EDV) and end-systolic (ESV) volumes increased less with combination therapy (EDV 8+/-4 mL; ESV 1+/-4 mL; P<0.01) than with candesartan alone (EDV 27+/-4 mL; ESV 18+/-3 mL) or enalapril alone (EDV 23+/-7 mL; ESV 14+/-6 mL). Blood pressure decreased with combination therapy (6+/-1/4+/-1 mm Hg) compared with candesartan or enalapril alone (P<0.05). Aldosterone decreased (P<0.05) with combination therapy (23.2+/-5.3 pg/mL) at 17 but not 43 weeks compared with candesartan (0.7+/-7.8 pg/mL) or enalapril (-0.8+/-11. 3 pg/mL). Brain natriuretic peptide decreased with combination therapy (5.8+/-2.7 pmol/L; P<0.01) compared with candesartan (4. 4+/-3.8 pmol/L) and enalapril alone (4.0+/-5.0 pmol/L). CONCLUSIONS: Candesartan alone was as effective, safe, and tolerable as enalapril. The combination of candesartan and enalapril was more beneficial for preventing left ventricular remodeling than either candesartan or enalapril alone. PMID- 10477531 TI - Origin of oscillatory kinetics of respiratory gas exchange in chronic heart failure. AB - BACKGROUND: Respiratory gas exchange measurements in patients with chronic heart failure (CHF) at rest and during exercise commonly reveal prominent slow oscillations in ventilation (V(E)), measured oxygen uptake (VO(2)), and carbon dioxide production (VCO(2)), whose origin is not clear. Voluntary simulation of periodic breathing (PB) in normals has been reported to generate a different pattern of oscillations in gas exchange from that seen in spontaneous PB. This necessitates hypothesizing that PB is caused by a primary oscillation in tissue metabolism or in cardiac output. METHODS AND RESULTS: We developed an automated method by which normal controls could be guided to breathe according to a PB pattern. The resultant metabolic oscillations closely matched those seen in spontaneous PB and had several interesting properties. At low workloads (including rest), the oscillations in VO(2) were as prominent as those in V(E) in both spontaneous PB (alpha(VO2)/alpha(VE)=0.92+/-0.04) and voluntary PB (0.93+/ 0.07). However, at increased workload, the oscillations in VO(2) because less prominent than those in V(E) in spontaneous PB (intermediate workload 0.63+/ 0.05, high workload 0.57+/-0.04; P<0.001) and voluntary PB (intermediate 0.66+/ 0.03, high 0.48+/-0.03; P<0.001). There was no difference in the relative size of metabolic oscillations between voluntary and spontaneous PB at matched workloads (P>0.05 at low, intermediate, and high workloads). Furthermore, VO(2) peaked before V(E) in both spontaneous and voluntary PB. This time delay varied from 6.4+/-0.4 s at low ventilation, to 11.3+/-0.9 s at high ventilation (P<0.0001). CONCLUSIONS: The magnitude and phase pattern of oscillations in gas exchange of spontaneous PB can be obtained by adequately matched voluntary PB. Therefore, the gas exchange features of PB are explicable by primary ventilatory oscillation. PMID- 10477532 TI - Prediction of coronary and cerebrovascular morbidity and mortality by direct continuous ambulatory blood pressure monitoring in essential hypertension. AB - BACKGROUND: The goal of this study was to assess the prognostic value of ambulatory versus clinic blood pressure measurement and to relate cardiovascular risk to ambulatory systolic and diastolic blood pressure levels. METHODS AND RESULTS: The study population consisted of 688 patients 51+/-11 years of age who had undergone pretreatment 24-hour intra-arterial ambulatory blood pressure monitoring on the basis of elevated clinic blood pressure. A total of 157 first events were recorded during a 9.2+/-4.1-year follow-up period. The predictive value of a regression model containing age, sex, race, body mass index, smoking, diabetes mellitus, fasting cholesterol level, and previous history of cardiovascular disease was significantly improved by the addition of any ambulatory systolic or diastolic blood pressure parameter (whether 24-hour, daytime, or nighttime mean) or pulse pressure, whereas inclusion of baseline clinic blood pressure variables did not enhance the prediction of events. The most predictive models contained the ambulatory systolic blood pressure parameters. In the model containing 24-hour mean ambulatory systolic blood pressure (P=0.001), age (P<0.001), male sex (P<0.001), South Asian origin (P=0.008), diabetes mellitus (P=0. 05), and previous cardiovascular disease (P<0.001) were additional independent predictors of events. Whereas 24-hour ambulatory systolic blood pressure was linearly related to the incidence of both coronary and cerebrovascular events, 24-hour ambulatory diastolic blood pressure exhibited a positive linear relationship with cerebrovascular events but a curvilinear relationship with coronary events. CONCLUSIONS: Ambulatory blood pressure is superior to clinic measurement for the assessment of cardiovascular risk; there is no reduction in coronary risk at lower levels of ambulatory diastolic blood pressure. PMID- 10477533 TI - Splicing mutations in KCNQ1: a mutation hot spot at codon 344 that produces in frame transcripts. AB - BACKGROUND: Long-QT syndrome is a monogenic disorder that produces cardiac arrhythmias and can lead to sudden death. At least 5 loci and 4 known genes exist in which mutations have been shown to be responsible for the disease. The potassium channel gene KCNQ1, previously named KVLQT1, on chromosome 11p15.5 is one of these. METHODS AND RESULTS: We initially analyzed one family using microsatellite markers and found linkage to KCNQ1. Mutation detection showed a G to C change in the last base of exon 6 (1032 G-->C) that does not alter the coded alanine. Restriction digest analysis in the family showed that only affected individuals carried the mutation. A previous report suggested that a G to A substitution at the same position may act as a splice mutation in KCNQ1, but no data was given to support this hypothesis nor was the transcription product identified. We have shown by reverse-transcription polymerase chain reaction that 2 smaller bands were produced for the KCNQ1 gene transcripts in addition to the normal-sized transcripts when lymphocytes of affected individuals were analyzed. Sequencing these transcripts showed a loss of exon 7 in one and exons 6 and 7 in the other, but an in-frame transcript was left in each instance. We examined other families in whom long-QT syndrome was diagnosed and found another unreported splice-site mutation, 922-1 G-->C, in the acceptor site of intron 5, and 2 of the previously reported 1032 G-->A mutations. All these showed a loss of exons 6 and 7 in the mutant transcripts, validating the proposal that a consensus sequence is affected in the exonic mutations and that the integrity of the base at position 1032 is essential for correct processing of the transcript. CONCLUSIONS: The 6 cases already reported in the literature with the 1032 G-->A transition, the novel 1032 G-->C transversion, and a recent G-->T transversion at the same base show that codon 344 is the second most frequently mutated after codon 341, suggesting at least two hotspots for mutations in KCNQ1. PMID- 10477534 TI - Aerobic exercise training can reverse age-related peripheral circulatory changes in healthy older men. AB - BACKGROUND: The age-related decline in maximal oxygen consumption is attenuated by habitual aerobic exercise. However, the relative effects of training on central and peripheral responses to exercise in older subjects are not known. The present study assessed the contribution of central and peripheral responses to the age-associated decline in peak oxygen consumption and compared the effect of exercise training in healthy older and younger subjects. METHODS AND RESULTS: Ten older and 13 younger men underwent invasive measurement of central and peripheral cardiovascular responses during an upright, staged cycle exercise test before and after a 3-month period of exercise training with cycle ergometry. At baseline, cardiac output and AV oxygen difference during exercise were significantly lower in older subjects. With training, the older and younger groups increased maximal oxygen consumption by 17.8% and 20.2%, respectively. Peak cardiac output was unchanged in both groups. Systemic AV oxygen difference increased 14.4% in the older group and 14.3% in the younger group and accounted for changes in peak oxygen consumption. Peak leg blood flow increased by 50% in older subjects, whereas the younger group showed no significant change. There was no change in peak leg oxygen extraction in the older group, but in the younger group, leg AV oxygen difference increased by 15.4%. CONCLUSIONS: These findings suggest that the age-related decline in maximal oxygen consumption results from a reversible deconditioning effect on the distribution of cardiac output to exercising muscle and an age-related reduction in cardiac output reserve. PMID- 10477535 TI - Raloxifene acutely relaxes rabbit coronary arteries in vitro by an estrogen receptor-dependent and nitric oxide-dependent mechanism. AB - BACKGROUND: Selective estrogen receptor modulators (SERMs) have been defined as compounds that display tissue specificity with regard to estrogenic effects. They appear to share the beneficial effects of estrogen on bone and lipids but are not associated with an increased risk of breast or uterine carcinoma. Estrogen relaxes coronary arteries and has long-term protective effects on the vascular system. The effect of SERMs on the coronary vasculature is unknown. We therefore investigated the effects of the SERM raloxifene on isolated rabbit coronary arteries. METHODS AND RESULTS: Rings of coronary artery from adult male and nonpregnant female New Zealand White rabbits were suspended in organ baths containing Krebs solution; isometric tension was then measured. Raloxifene induced coronary arterial relaxation in male and female coronary arteries by an endothelium-dependent and estrogen receptor-dependent mechanism involving nitric oxide. Raloxifene also had a direct calcium antagonistic effect on the coronary myocyte. Estrogen, however, induced only endothelium-independent coronary arterial relaxation. The endothelium-dependent component of relaxation induced by raloxifene 10(-6) mol/L resulted in almost 100% (79+/-7% versus 43+/-3%, P<0.001) more relaxation than that induced by estrogen 10(-6) mol/L. CONCLUSIONS: These data demonstrate that raloxifene has vascular relaxing properties. The surprising finding is that the receptor-dependent effects via the endothelium are observed in coronary arteries from both male and female animals. PMID- 10477536 TI - Treatment of experimental viral myocarditis with interleukin-10. AB - BACKGROUND: The T helper cell type 2-associated cytokine interleukin (IL)-10 has a variety of immunomodulatory properties. However, the effects of the cytokine on viral myocarditis remain unclear. METHODS AND RESULTS: We studied the effects of recombinant human IL-10 (rhIL-10) fully active on mouse cells in a murine experimental model of acute viral myocarditis caused by the encephalomyocarditis virus (EMCV). Four-week-old DBA/2 mice were inoculated with EMCV (day 0). rhIL-10 (10 microg/mouse) was administered once daily, starting on day 0, and control mice received vehicle only. Survival rates were determined on day 14. Myocardial histopathology, cytokine levels in the heart by ELISA assay, and myocardial virus concentration were examined on day 6, and the expression levels of myocardial inducible nitric oxide synthase (iNOS) mRNA were measured by competitive polymerase chain reaction. The 14-day survival in mice treated with rhIL-10 was significantly higher (80%) than in the control group (30%, n=10 in each, P<0.05). rhIL-10 treatment significantly attenuated myocardial lesions and suppressed tumor necrosis factor-alpha and IL-2 in the heart. rhIL-10 treatment had little effect on myocardial virus concentration. The expression levels of myocardial iNOS mRNA were significantly decreased in the group treated with rhIL-10 (8.6+/ 4.7 amol/mg total RNA in treated versus 26.5+/-7.1 amol/mg total RNA in control mice, P<0.05). CONCLUSIONS: These findings provide new insights into the in vivo effects of IL-10 on viral infection and suggest a therapeutic effect of IL-10 on viral myocarditis. PMID- 10477537 TI - Endothelial dysfunction and collagen accumulation: two independent factors for restenosis and constrictive remodeling after experimental angioplasty. AB - BACKGROUND: Constrictive remodeling plays a prominent role in restenosis after balloon angioplasty, but its regulation remains unclear. Because endothelial dysfunction and changes in extracellular matrix have been reported after angioplasty, this study was designed to simultaneously evaluate endothelial function and collagen and elastin changes after restenosis and arterial remodeling. METHODS AND RESULTS: Atherosclerosis was induced in femoral arteries of 22 New Zealand White rabbits by air-desiccation and a high-cholesterol diet. One month later, angioplasty was performed. Histomorphometry and in vitro assessment of endothelial function were performed 4 weeks after angioplasty. Restenosis correlated with constrictive remodeling (r=0.60, P=0.01) but not with neointimal growth (r=-0.06, P=0.79). Restenosis correlated with an impaired relaxation to acetylcholine (ACh; r=0.61, P=0.02) but not with the response to the endothelium-independent vasodilator sodium nitroprusside (r=-0.25, P=0.40). Restenosis correlated positively with collagen accumulation (r=0.69, P=0.004) and inversely with elastin density (r=-0.48, P=0.05). Relaxations to ACh were significantly more decreased in arteries with constrictive remodeling than in those with enlargement remodeling (3.7+/-7.9% versus 35.5+/-15.0%, P=0.04). Neointimal collagen density was significantly higher in arteries with constrictive remodeling than in those with enlargement remodeling (34.5+/-4.5% versus 18.2+/-4.7%, P=0.03). Endothelial function and collagen and elastin density were independent predictors of restenosis in the study. CONCLUSIONS: These results demonstrate that the severity of restenosis after angioplasty correlated with both defective endothelium-dependent relaxation and increased collagen density. PMID- 10477538 TI - Involvement of cardiotrophin-1 in cardiac myocyte-nonmyocyte interactions during hypertrophy of rat cardiac myocytes in vitro. AB - BACKGROUND: The mechanism responsible for cardiac hypertrophy is currently conceptualized as having 2 components, mediated by cardiac myocytes and nonmyocytes, respectively. The interaction between myocytes and nonmyocytes via growth factors and/or cytokines plays an important role in the development of cardiac hypertrophy. We found that cardiac myocytes showed hypertrophic changes when cocultured with cardiac nonmyocytes. Cardiotrophin-1 (CT-1), a new member of the interleukin-6 family of cytokines, was identified by its ability to induce hypertrophic response in cardiac myocytes. In this study, we used the in vitro coculture system to examine how CT-1 is involved in the interaction between cardiac myocytes and nonmyocytes during the hypertrophy process. METHODS AND RESULTS: RNase protection assay revealed that CT-1 mRNA levels were 3. 5 times higher in cultured cardiac nonmyocytes than in cultured cardiac myocytes. We developed anti-CT-1 antibodies and found that they significantly inhibited the increased atrial and brain natriuretic peptide secretion and protein synthesis characteristic of hypertrophic changes of myocytes in the coculture. In addition, non-myocyte-conditioned medium rapidly elicited tyrosine phosphorylation of STAT3 and induced an increase in natriuretic peptide secretion and protein synthesis in cultured cardiac myocytes; these effects were partially suppressed by anti-CT-1 antibodies. Finally, the hypertrophic effects of CT-1 and endothelin-1, which we had previously implicated in the hypertrophic activity in the coculture, were additive in cardiac myocytes. CONCLUSIONS: These results show that CT-1 secreted from cardiac nonmyocytes is significantly involved in the hypertrophic changes of cardiac myocytes in the coculture and suggest that CT-1 is an important local regulator in the process of cardiac hypertrophy. PMID- 10477539 TI - Relation of atrial refractoriness to upper and lower limits of vulnerability for atrial fibrillation/flutter following implantable ventricular defibrillator shocks. AB - BACKGROUND: Implantable ventricular cardioverter defibrillator (ICD) shocks can cause atrial fibrillation/flutter (AF). This study investigated the pathogenesis of AF after ICD shocks in a canine model. METHODS AND RESULTS: The study was conducted in 8 dogs. In 5 dogs (group 1), truncated exponential (8 ms, 78% tilt) monophasic and biphasic shocks were delivered through a bipolar epicardial (patch) or endocardial lead. After the last S1 of atrial pacing at a cycle length of 350 ms, shocks of 0.1 to 7.6 A (0.005 to 27.7 J) were delivered, timed to the atrial effective refractory period (AERP). Ventricular defibrillation thresholds were also determined. In 3 dogs (group 2), the effect of the open versus closed chest technique on AF induction was tested in the endocardial biphasic shock configuration. AF was induced in all 8 dogs and in all waveforms and configurations. Mean AF duration was 11.5+/-6 s, with a mean ventricular rate of 184+/-37 bpm. Ventricular shocks could induce AF only if they were timed between an AERP of -60 to 40 ms, -40 to 60 ms, -40 to 60 ms, and -20 to 60 ms in the epicardial monophasic, epicardial biphasic, endocardial monophasic, and endocardial biphasic configurations, respectively. The mean+/-SD of the upper limit of vulnerability (ULV) for AF induction (in J) was 5. 2+/-0.6, 3.5+/-0.4, 5.2+/-1.2, and 2.5+/-0.1 for the epicardial monophasic, epicardial biphasic, endocardial monophasic, and endocardial biphasic configurations, respectively (P<0.05). The lower limit of vulnerability (LLV) was 0.8+/-0.1, 0.8+/-0.1, 0.9+/ 0, and 0.6+/-0 for the epicardial monophasic, epicardial biphasic, endocardial monophasic, and endocardial biphasic configurations, respectively (P=NS). The ventricular defibrillation threshold (in J) for all wave forms and configurations was higher than the ULV (P<0. 05). CONCLUSIONS: (1) An atrial LLV and ULV exist for ventricular ICD shock-induced AF; (2) the shock-induced AF is related to both shock intensity and its timing to AERP; and (3) avoiding this atrial window of vulnerability may minimize the risk of post-ICD shock AF. PMID- 10477540 TI - Aortocoronary bypass surgery in a patient with aortic calcification. PMID- 10477541 TI - Diabetes mellitus: a major risk factor for cardiovascular disease. A joint editorial statement by the American Diabetes Association; The National Heart, Lung, and Blood Institute; The Juvenile Diabetes Foundation International; The National Institute of Diabetes and Digestive and Kidney Diseases; and The American Heart Association. PMID- 10477542 TI - Diabetes and cardiovascular disease: a statement for healthcare professionals from the American Heart Association. PMID- 10477543 TI - Cholesterol embolism as a complication of aortic dissection. PMID- 10477544 TI - Interference with the signaling capacity of CC chemokine receptor 5 can compromise its role as an HIV-1 entry coreceptor in primary T lymphocytes. PMID- 10477545 TI - The B-oligomer of pertussis toxin deactivates CC chemokine receptor 5 and blocks entry of M-tropic HIV-1 strains. AB - Infection of target cells by HIV-1 requires initial binding interactions between the viral envelope glycoprotein gp120, the cell surface protein CD4, and one of the members of the seven-transmembrane G protein-coupled chemokine receptor family. Most primary isolates (R5 strains) use chemokine receptor CCR5, but some primary syncytium-inducing, as well as T cell line-adapted, strains (X4 strains) use the CXCR4 receptor. Signaling from both CCR5 and CXCR4 is mediated by pertussis toxin (PTX)-sensitive G(i) proteins and is not required for HIV-1 entry. Here, we show that the PTX holotoxin as well as its binding subunit, B oligomer, which lacks G(i)-inhibitory activity, blocked entry of R5 but not X4 strains into primary T lymphocytes. Interestingly, B-oligomer inhibited virus production by peripheral blood mononuclear cell cultures infected with either R5 or X4 strains, indicating that it can affect HIV-1 replication at both entry and post-entry levels. T cells treated with B-oligomer did not initiate signal transduction in response to macrophage inflammatory protein (MIP)-1beta or RANTES (regulated upon activation, normal T cell expressed and secreted); however, cell surface expression of CCR5 and binding of MIP-1beta or HIV-1 to such cells were not impaired. The inhibitory effect of B-oligomer on signaling from CCR5 and on entry of R5 HIV-1 strains was reversed by protein kinase C (PKC) inhibitors, indicating that B-oligomer activity is mediated by signaling events that involve PKC. B-oligomer also blocked cocapping of CCR5 and CD4 induced by R5 HIV-1 in primary T cells, but did not affect cocapping of CXCR4 and CD4 after inoculation of the cultures with X4 HIV-1. These results suggest that the B-oligomer of PTX cross-deactivates CCR5 to impair its function as a coreceptor for HIV-1. PMID- 10477546 TI - Alteration of interleukin 4 production results in the inhibition of T helper type 2 cell-dominated inflammatory bowel disease in T cell receptor alpha chain deficient mice. AB - T cell receptor alpha chain-deficient (TCR-alpha(-/-)) mice are known to spontaneously develop inflammatory bowel disease (IBD). The colitis that develops in these mice is associated with increased numbers of T helper cell (Th)2-type CD4(+)TCR-betabeta (CD4(+)betabeta) T cells producing predominantly interleukin (IL)-4. To investigate the role of these Th2-type CD4(+)betabeta T cells, we treated TCR-alpha(-/-) mice with anti-IL-4 monoclonal antibody (mAb). Approximately 60% of TCR-alpha(-/-) mice, including those treated with mock Ab and those left untreated, spontaneously developed IBD. However, anti-IL-4 mAb treated mice exhibited no clinical or histological signs of IBD, and their levels of mucosal and systemic Ab responses were lower than those of mock Ab-treated mice. Although TCR-alpha(-/-) mice treated with either specific or mock Ab developed CD4(+)betabeta T cells, only those treated with anti-IL-4 mAb showed a decrease in Th2-type cytokine production at the level of mRNA and protein and an increase in interferon gamma-specific expression. These findings suggest that IL 4-producing Th2-type CD4(+)betabeta T cells play a major immunopathological role in the induction of IBD in TCR-alpha(-/-) mice, a role that anti-IL-4 mAb inhibits by causing Th2-type CD4(+)betabeta T cells to shift to the Th1 type. PMID- 10477547 TI - Distinct role of antigen-specific T helper type 1 (Th1) and Th2 cells in tumor eradication in vivo. AB - The role of T helper type 1 (Th1) and Th2 cells in tumor immunity was investigated using Th cells induced from ovalbumin (OVA)-specific T cell receptor transgenic mice. Although Th1 cells exhibited stronger cytotoxicity than Th2 cells, both cell types completely eradicated tumors when transferred into mice bearing A20 tumor cells transfected with the OVA gene (A20-OVA). Th1 cells eradicated the tumor mass by inducing cellular immunity, whereas Th2 cells destroyed the tumor by inducing tumor necrosis. Both Th1 and Th2 cells required CD8(+) T cells to eliminate tumors, and neither of these cells were able to completely eliminate A20-OVA tumors from T and B cell-deficient RAG2(-/-) mice. Mice cured from tumors by Th1 and Th2 cell therapy rejected A20-OVA upon rechallenge, but CD8(+) cytotoxic T lymphocytes were induced only from spleen cells prepared from cured mice by Th1 cell therapy. Moreover, we demonstrated that Th1 and Th2 cells used distinct adhesion mechanisms during tumor eradication: the leukocyte function-associated antigen (LFA)-1-dependent cell cell adhesion step was essential for Th1 cell therapy, but not for Th2 cell therapy. These findings demonstrated for the first time the distinct role of antigen-specific Th1 and Th2 cells during eradication of established tumors in vivo. PMID- 10477548 TI - The requirement of membrane lymphotoxin for the presence of dendritic cells in lymphoid tissues. AB - Although several cytokines, including tumor necrosis factor (TNF), can promote the growth of dendritic cells (DCs) in vitro, the cytokines that naturally regulate DC development and function in vivo have not been well defined. Here, we report that membrane lymphotoxin (LT), instead of TNF, regulates the migration of DCs in the spleen. LTalpha(-/-) mice, lacking membrane LTalpha/beta and LTalpha(3), show markedly reduced numbers of DCs in the spleen. Unlike wild-type mice and TNF(-/-) mice that have densely clustered DCs in the T cell zone and around the marginal zone, splenic DCs in LTalpha(-/-) mice are randomly distributed. The reduced number of DCs in lymphoid tissues of LTalpha(-/-) mice is associated with an increased number of DCs in nonlymphoid tissues. The number of splenic DCs in LTalpha(-/-) mice is restored when additional LT-expressing cells are provided. Blocking membrane LTalpha/beta in wild-type mice markedly diminishes the accumulation of DCs in lymphoid tissues. These data suggest that membrane LT is an essential ligand for the presence of DCs in the spleen. Mice deficient in TNF receptor, which is the receptor for both soluble LTalpha(3) and TNF-alpha(3) trimers, have normal numbers of DCs. However, LTbetaR(-/-) mice show reduced numbers of DCs, similar to the mice lacking membrane LT alpha/beta. Taken together, these results support the notion that the signaling via LTbetaR by membrane LTalpha/beta is required for the presence of DCs in lymphoid tissues. PMID- 10477549 TI - Autoantigen-specific B cell activation in Fas-deficient rheumatoid factor immunoglobulin transgenic mice. AB - In systemic autoimmune disease, self-tolerance fails, leading to autoantibody production. A central issue in immunology is to understand the origins of activated self-reactive B cells. We have used immunoglobulin (Ig) transgenic mice to investigate the regulation of autoreactive B cells with specificity for self IgG2a (the rheumatoid factor [RF] specificity) to understand how normal mice regulate RF autoantibodies and how this fails in autoimmune mice. We previously showed that normal mice do not tolerize the AM14 RF clone, nor do they appear to activate it. Here we show that in Fas-deficient autoimmune mice, the picture is quite different. RF B cells are activated to divide and secrete, but only when the autoantigen is present. Thus, B cells that are ignored rather than anergized in normal mice can be stimulated to produce autoantibody in Fas-deficient mice. This demonstrates a novel developmental step at which intact Fas-Fas ligand signaling is required to regulate B cells in order to prevent autoimmunity. These data also establish the relevance of ignorant self-specific B cells to autoantibody production in disease and prove that in the case of the RF specificity, the nominal autoantigen IgG2a is the driving autoantigen in vivo. PMID- 10477550 TI - An expanded peripheral T cell population to a cytotoxic T lymphocyte (CTL) defined, melanocyte-specific antigen in metastatic melanoma patients impacts on generation of peptide-specific CTLs but does not overcome tumor escape from immune surveillance in metastatic lesions. AB - It is not known if immune response to T cell-defined human histocompatibility leukocyte antigen (HLA) class I-restricted melanoma antigens leads to an expanded peripheral pool of T cells in all patients, affects cytotoxic T lymphocyte (CTL) generation, and correlates with anti-tumor response in metastatic lesions. To this end, a limiting dilution analysis technique was developed that allowed us to evaluate the same frequency of peptide-specific T cells as by staining T cells with HLA-peptide tetrameric complexes. In four out of nine patients, Melan-A/Mart 1(27-35)-specific CTL precursors (CTLp) were >/=1/2,000 peripheral blood lymphocytes and found mostly or only in the CD45RO(+) memory T cell subset. In the remaining five patients, a low (<1/40,000) peptide-specific CTLp frequency was measured, and the precursors were only in the CD45RA(+) naive T cell subset. Evaluation of CTL effector frequency after bulk culture indicated that peptide specific CTLs could be activated in all patients by using professional antigen presenting cells as dendritic cells, but CTLp frequency determined the kinetics of generation of specificity and the final number of effectors as evaluated by both limiting dilution analysis and staining with HLA-A*0201-Melan-A/Mart-1 tetrameric complexes. Immunohistochemical analysis of 26 neoplastic lesions from the nine patients indicated absence of tumor regression in most instances, even in patients with an expanded peripheral T cell pool to Melan-A/Mart-1 and whose neoplastic lesions contained a high frequency of tetramer-positive Melan-A/Mart-1 specific T cells. Furthermore, frequent lack of a "brisk" or "nonbrisk" CD3(+)CD8(+) T cell infiltrate or reduced/absent Melan-A/Mart-1 expression in several lesions and lack of HLA class I antigens were found in some instances. Thus, expansion of peripheral immune repertoire to Melan-A/Mart-1 takes place in some metastatic patients and leads to enhanced CTL induction after antigen presenting cell-mediated selection, but, in most metastatic lesions, it does not overcome tumor escape from immune surveillance. PMID- 10477551 TI - A novel element upstream of the Vgamma2 gene in the murine T cell receptor gamma locus cooperates with the 3' enhancer to act as a locus control region. AB - Transgenic expression constructs were employed to identify a cis-acting transcription element in the T cell receptor (TCR)-gamma locus, called HsA, between the Vgamma5 and Vgamma2 genes. In constructs lacking the previously defined enhancer (3'E(Cgamma1)), HsA supports transcription in mature but not immature T cells in a largely position-independent fashion. 3'E(Cgamma1), without HsA, supports transcription in immature and mature T cells but is subject to severe position effects. Together, the two elements support expression in immature and mature T cells in a copy number-dependent, position-independent fashion. Furthermore, HsA was necessary for consistent rearrangement of transgenic recombination substrates. These data suggest that HsA provides chromatin-opening activity and, together with 3'E(Cgamma1), constitutes a T cell specific locus control region for the TCR-gamma locus. PMID- 10477552 TI - Abnormal chemokine-induced responses of immature and mature hematopoietic cells from motheaten mice implicate the protein tyrosine phosphatase SHP-1 in chemokine responses. AB - Chemokines regulate a number of biological processes, including trafficking of diverse leukocytes and proliferation of myeloid progenitor cells. SHP-1 (Src homology 2 domain tyrosine phosphatase 1), a phosphotyrosine phosphatase, is considered an important regulator of signaling for a number of cytokine receptors. Since specific tyrosine phosphorylation of proteins is important for biological activities induced by chemokines, we examined the role of SHP-1 in functions of chemokines using viable motheaten (me(v)/me(v)) mice that were deficient in SHP-1. Chemotactic responses to stromal call-derived factor 1 (SDF 1), a CXC chemokine, were enhanced with bone marrow myeloid progenitor cells as well as macrophages, T cells, and B cells from me(v)/me(v) versus wild-type (+/+) mice. SDF-1-dependent actin polymerization and activation of mitogen-activated protein kinases were also greater in me(v)/me(v) versus +/+ cells. In contrast, immature subsets of me(v)/me(v) bone marrow myeloid progenitors were resistant to effects of a number of chemokines that suppressed proliferation of +/+ progenitors. These altered chemokine responses did not appear to be due to enhanced expression of CXCR4 or lack of chemokine receptor expression. However, expression of some chemokine receptors (CCR1, CCR2, CCR3, and CXCR2) was significantly enhanced in me(v)/me(v) T cells. Our results implicate SHP-1 involvement in a number of different chemokine-induced biological activities. PMID- 10477553 TI - Somatic mutation and light chain rearrangement generate autoimmunity in anti single-stranded DNA transgenic MRL/lpr mice. AB - Antibodies to single-stranded (ss)DNA are expressed in patients with systemic lupus erythematosus and in lupus-prone mouse models such as the MRL/Mp-lpr/lpr (MRL/lpr) strain. In nonautoimmune mice, B cells bearing immunoglobulin site directed transgenes (sd-tgs) that code for anti-ssDNA are functionally silenced. In MRL/lpr autoimmune mice, the same sd-tgs are expressed in peripheral B cells and these autoantibodies gain the ability to bind other autoantigens such as double-stranded DNA and cell nuclei. These new specificities arise by somatic mutation of the anti-ssDNA sd-tgs and by secondary light chain rearrangement. Thus, B cells that in normal mice are anergic can be activated in MRL/lpr mice, which can lead to the generation of pathologic autoantibodies. In this paper, we provide the first direct evidence for peripheral rearrangement in vivo. PMID- 10477554 TI - High frequencies of naive Melan-A/MART-1-specific CD8(+) T cells in a large proportion of human histocompatibility leukocyte antigen (HLA)-A2 individuals. AB - Using fluorescent HLA-A*0201 tetramers containing the immunodominant Melan-A/MART 1 (Melan-A) tumor-associated antigen (Ag), we previously observed that metastatic lymph nodes of melanoma patients contain high numbers of Ag-experienced Melan-A specific cytolytic T lymphocytes (CTLs). In this paper, we enumerated and characterized ex vivo Melan-A-specific cells in peripheral blood samples from both melanoma patients and healthy individuals. High frequencies (>/=1 in 2,500 CD8(+) T cells) of Melan-A-specific cells were found in 10 out of 13 patients, and, surprisingly, in 6 out of 10 healthy individuals. Virtually all Melan-A specific cells from 6 out of 6 healthy individuals and from 7 out of 10 patients displayed a naive CD45RA(hi)/RO(-) phenotype, whereas variable proportions of Ag experienced CD45RA(lo)/RO(+) Melan-A-specific cells were observed in the remaining 3 patients. In contrast, ex vivo influenza matrix-specific CTLs from all individuals exhibited a CD45RA(lo)/RO(+) memory phenotype as expected. Ag specificity of tetramer-sorted A2/Melan-A(+) cells from healthy individuals was confirmed after mitogen-driven expansion. Likewise, functional limiting dilution analysis and interferon gamma ELISPOT assays independently confirmed that most of the Melan-A-specific cells were not Ag experienced. Thus, it appears that high frequencies of naive Melan-A-specific CD8(+) T cells can be found in a large proportion of HLA-A*0201(+) individuals. Furthermore, as demonstrated for one patient followed over time, dramatic phenotype changes of circulating Melan-A specific cells can occur in vivo. PMID- 10477555 TI - Mycobacterium tuberculosis expresses a novel pH-dependent divalent cation transporter belonging to the Nramp family. AB - Mammalian natural resistance-associated macrophage protein (Nramp) homologues are important determinants of susceptibility to infection by diverse intracellular pathogens including mycobacteria. Eukaryotic Nramp homologues transport divalent cations such as Fe(2+), Mn(2+), Zn(2+), and Cu(2+). Mycobacterium tuberculosis and Mycobacterium bovis (bacillus Calmette-Guerin [BCG]) also encode an Nramp homologue (Mramp). RNA encoding Mramp induces approximately 20-fold increases in (65)Zn(2+) and (55)Fe(2+) uptake when injected into Xenopus laevis oocytes. Transport is dependent on acidic extracellular pH and is maximal between pH 5.5 and 6.5. Mramp-mediated (65)Zn(2+) and (55)Fe(2+) transport is abolished by an excess of Mn(2+) and Cu(2+), confirming that Mramp interacts with a broad range of divalent transition metal cations. Using semiquantitative reverse transcription PCR, we show that Mramp mRNA levels in M. tuberculosis are upregulated in response to increases in ambient Fe(2+) and Cu(2+) between <1 and 5 microM concentrations and that this upregulation occurs in parallel with mRNA for y39, a putative metal-transporting P-type ATPase. Using a quantitative ratiometric PCR technique, we demonstrate a fourfold decrease in Mramp/y39 mRNA ratios from organisms grown in 5-70 microM Cu(2+). M. bovis BCG cultured axenically and within THP-1 cells also expresses mRNA encoding Mramp. Mramp exemplifies a novel prokaryotic class of metal ion transporter. Within phagosomes, Mramp and Nramp1 may compete for the same divalent cations, with implications for intracellular survival of mycobacteria. PMID- 10477557 TI - Studies in B7-deficient mice reveal a critical role for B7 costimulation in both induction and effector phases of experimental autoimmune encephalomyelitis. AB - The importance of B7 costimulation in regulating T cell expansion and peripheral tolerance suggests that it may also play a significant regulatory role in the development of autoimmune disease. It is unclear whether B7 costimulation is involved only in the expansion of autoreactive T cells in the periphery, or if it is also required for effector activation of autoreactive T cells in the target organ for mediating tissue injury and propagating autoimmune disease. In this study, the role of B7-CD28 costimulation and the relative importance of B7 costimulators for the induction and effector phases of experimental autoimmune encephalomyelitis (EAE) induced by myelin oligodendrocyte glycoprotein (MOG) peptide were examined. Wild-type, B7-1/B7-2-deficient mice, or CD28-deficient C57BL/6 mice were immunized with MOG 35-55 peptide. Mice lacking both B7-1 and B7 2 or CD28 showed no or minimal clinical signs of EAE and markedly reduced inflammatory infiltrates in the brain and spinal cord. However, mice lacking either B7-1 or B7-2 alone developed clinical and pathologic EAE that was comparable to EAE in wild-type mice, indicating overlapping functions for B7-1 and B7-2. Resistance to EAE was not due to a lack of induction of T helper type 1 (Th1) cytokines, since T cells from B7-1/B7-2(-/-) mice show reduced proliferative responses, but greater interferon gamma production compared with T cells from wild-type mice. To study the role of B7 molecules in the effector phase of the disease, MOG 35-55-specific T lines were adoptively transferred into the B7-1/B7-2(-/-) and wild-type mice. Clinical and histologic EAE were markedly reduced in B7-1/B7-2(-/-) compared with wild-type recipient mice. These results demonstrate that B7 costimulation has critical roles not only in the initial activation and expansion of MOG-reactive T cells, but also in the effector phase of encephalitogenic T cell activation within the central nervous system. PMID- 10477556 TI - Measuring recent thymic emigrants in blood of normal and HIV-1-infected individuals before and after effective therapy. AB - The role of the thymus in HIV-1 pathogenesis remains unclear. We developed an assay to quantify the number of recent thymic emigrants in blood based on the detection of a major excisional DNA byproduct (termed alpha1 circle) of T cell receptor rearrangement. By studying 532 normal individuals, we found that alpha1 circle numbers in blood remain high for the first 10-15 yr of life, a sharp drop is seen in the late teen years, and a gradual decline occurs thereafter. Compared with age-matched uninfected control individuals, alpha1 circle numbers in HIV-1 infected adults were significantly reduced; however, there were many individuals with normal alpha1 circle numbers. In 74 individuals receiving highly active antiretroviral therapy, we found no appreciable effect on alpha1 circle numbers in those whose baseline values were already within the normal range, but significant increases were observed in those with a preexisting impairment. The increases in alpha1 circle numbers were, however, numerically insufficient to account for the rise in levels of naive T lymphocytes. Overall, it is difficult to invoke thymic regenerative failure as a generalized mechanism for CD4 lymphocyte depletion in HIV-1 infection, as alpha1 circle numbers are normal in a substantial subset of HIV-1-infected individuals. PMID- 10477558 TI - N-formylpeptides induce two distinct concentration optima for mouse neutrophil chemotaxis by differential interaction with two N-formylpeptide receptor (FPR) subtypes. Molecular characterization of FPR2, a second mouse neutrophil FPR. AB - The N-formylpeptide receptor (FPR) is a G protein-coupled receptor that mediates mammalian phagocyte chemotactic responses to bacterial N-formylpeptides. Here we show that a mouse gene named Fpr-rs2 encodes a second N-formylpeptide receptor subtype selective for neutrophils which we have provisionally named FPR2. The prototype N-formylpeptide fMLF induced calcium flux and chemotaxis in human embryonic kidney (HEK) 293 cells stably transfected with FPR2. The EC(50)s, approximately 5 microM for calcium flux and chemotaxis, were approximately 100 fold greater than the corresponding values for mouse FPR-transfected HEK 293 cells. Consistent with this, fMLF induced two distinct concentration optima for chemotaxis of normal mouse neutrophils, but only the high concentration optimum for chemotaxis of neutrophils from FPR knockout mice. Based on these data, we hypothesize that high- and low-affinity N-formylpeptide receptors, FPR and FPR2, respectively, may function in vivo as a relay mediating neutrophil migration through the high and low concentration portions of N-formylpeptide gradients. PMID- 10477559 TI - Cutting edge: SIV Nef protein utilizes both leucine- and tyrosine-based protein sorting pathways for down-regulation of CD4. AB - The Nef protein is unique to primate lentiviruses and is closely linked to accelerated pathogenesis in both human and monkey hosts. Nef acts to down regulate CD4 and MHC class I, two receptors important for immune function. A recent report demonstrated the presence of two tyrosine motifs in SIV Nef that contribute to its ability to down-regulate CD4 and to associate with clathrin adaptors. These tyrosine motifs are not present in HIV-1 Nef, which instead utilizes a leucine-based motif for its down-regulation of CD4. We now report that SIV Nef also contains a conserved leucine-based motif that contributes to CD4 down-regulation, functions to stimulate internalization, and contributes to the association of SIV Nef with clathrin adaptors AP-1 and AP-2. These results demonstrate that SIV Nef differs from HIV-1 Nef by its ability to use two parallel pathways of the protein-sorting machinery based on either tyrosine or leucine motifs. PMID- 10477560 TI - A dominant role for the thymus and MHC genes in determining the peripheral CD4/CD8 T cell ratio in the rat. AB - During their development, immature CD4CD8 double positive thymocytes become committed to either the CD4 or CD8 lineage. The final size of the peripheral CD4 and CD8 T cell compartments depends on thymic output and on the differential survival and proliferation of the respective T cell subsets in the periphery. Our results reveal that the development of the distinct peripheral CD4/CD8 T cell ratio between Lewis and Brown Norway rats originates in the thymus and, as shown by the use of radiation bone marrow chimeras, is determined by selection on radio resistant stromal cells. Furthermore, this difference is strictly correlated with the MHC haplotype and is the result of a reduction in the absolute number of CD8 T cells in Brown Norway rats. These data suggest that the distinct CD4/CD8 T cell ratio between these two rat strains is the consequence of differential interactions of the TCR/CD8 coreceptor complex with the respective MHC class I haplotypes during selection in the thymus. PMID- 10477561 TI - Role of the stress kinase pathway in signaling via the T cell costimulatory receptor 4-1BB. AB - 4-1BB is a member of the TNFR superfamily expressed on activated CD4+ and CD8+ T cells. 4-1BB can costimulate IL-2 production by resting primary T cells independently of CD28 ligation. In this study, we report signaling events following 4-1BB receptor aggregation using an Ak-restricted costimulation dependent T cell hybridoma, C8.A3. Aggregation of 4-1BB on the surface of C8.A3 cells induces TNFR-associated factor 2 recruitment, which in turn recruits and activates apoptosis signal-regulating kinase-1, leading to downstream activation of c-Jun N-terminal/stress-activated protein kinases (JNK/SAPK). 4-1BB ligation also enhances anti-CD3-induced JNK/SAPK activation in primary T cells. Overexpression of a catalytically inactive form of apoptosis signal-regulating kinase-1 in C8.A3 T cells interferes with activation of the SAPK cascade and with IL-2 secretion, consistent with a critical role for JNK/SAPK activation in 4-1BB dependent IL-2 production. Given the ability of both CD28 and 4-1BB to induce JNK/SAPK activation, we asked whether hyperosmotic shock, another inducer of this cascade, could function to provide a costimulatory signal to T cells. Osmotic shock of resting primary T cells in conjunction with anti-CD3 treatment was found to costimulate IL-2 production by the T cells, consistent with a pivotal role for JNK/SAPK in T cell costimulation. PMID- 10477562 TI - Differential coupling of second signals for cytotoxicity and proliferation in CD8+ T cell effectors: amplification of the lytic potential by B7. AB - The role of second signals delivered through B7/CD28 interactions in T cell activation is well documented. However, once CTLs are elicited, TCR-mediated cytotoxicity appears to be uncoupled from the requirement for costimulatory signals. In this study, we show an uncoupling across a broad range of concentrations of peptide, thus demonstrating that cytolysis is a TCR-mediated response that is fully independent of costimulatory signals. However, the same T cell effectors remain fully responsive to B7 engagement, which is able to amplify Ag-mediated proliferation and cytolytic capacity. B7 expression by targets results in an IL-2-mediated proliferative expansion of the effectors concurrent with the elimination of the targets. Thus, costimulation of effectors results in a vast expansion in lytic units over time, which does not occur in the absence of IL-2 or B7. Both TCR-derived and second signals appear to be necessary to achieve this result. These results suggest that B7-expressing APC or a cohort of IL-2 producing helper cells would functionally extend the duration and effectiveness of the cytotoxic response occurring in localized immune responses. PMID- 10477563 TI - Intracellular signaling of gp34, the OX40 ligand: induction of c-jun and c-fos mRNA expression through gp34 upon binding of its receptor, OX40. AB - We investigated the intracellular signaling events of OX40 ligand (gp34), a member of the TNF family. To elucidate the intracellular signaling via gp34, we prepared a model system in which a human gp34-transfected mouse epithelial cell line was stimulated with a recombinant soluble form of OX40. We demonstrated that OX40 binding resulted in increase in c-jun and c-fos mRNA levels in this transfectant by Northern blot analysis, which was blocked by the pretreatment with anti-gp34 Ab. The studies with various gp34 deletion mutants showed that the cytoplasmic portion including the amino acid sequence 16-21 (RPRFER) was required for the induction of c-jun and c-fos mRNA expression. Furthermore, OX40 binding induced c-jun mRNA expression also in HUVECs, which in our previous study have been shown to express gp34 and interact with activated T cells through the OX40/gp34 pathway. On the other hand, c-fos mRNA was detectable neither in unstimulated HUVECs nor in gp34-stimulated HUVECs. These results indicate that the OX40/gp34 system generates two-way signals and may elicit biological effects on vascular endothelial cells. PMID- 10477564 TI - Involvement of the SHP-1 tyrosine phosphatase in regulation of T cell selection. AB - The selection events shaping T cell development in the thymus represent the outcome of TCR-driven intracellular signaling cascades evoked by Ag receptor interaction with cognate ligand. In view of data indicating TCR-evoked thymocyte proliferation to be negatively modulated by the SHP-1 tyrosine phosphatase, a potential role for SHP-1 in regulating selection processes was investigated by analysis of T cell development in H-Y TCR transgenic mice rendered SHP-1 deficient by introduction of the viable motheaten mutation or a dominant negative SHP-1-encoding transgene. Characterization of thymocyte and peripheral T cell populations in H-Y TCR-viable motheaten mice revealed TCR-evoked proliferation as well as the positive and negative selection of H-Y-specific thymocytes to be enhanced in these mice, thus implicating SHP-1 in the negative regulation of each of these processes. T cell selection processes were also augmented in H-Y TCR mice carrying a transgene driving lymphoid-restricted expression of a catalytically inert, dominant-negative form of SHP-1. SHP-1-negative effects on thymocyte TCR signaling were not influenced by co-cross-linking of the CD28 costimulatory and/or CTLA-4 inhibitory receptors and appear, accordingly, to be realized independently of these comodulators. These observations indicate that SHP-1 raises the signaling threshold required for both positive and negative selection and reveal the inhibitory effects of SHP-1 on TCR signaling to be cell autonomous. The demonstrated capacity for SHP-1 to inhibit TCR-evoked proliferation and selection indicate SHP-1 modulatory effects on the magnitude of TCR-generated signal to be a key factor in determining the cellular consequences of TCR-ligand interaction. PMID- 10477565 TI - Peptide affinity and concentration affect the sensitivity of M3-restricted CTLs induced in vitro. AB - In vitro stimulation of mouse splenocytes with hemagglutinin (HA) 173-190, a peptide derived from influenza virus hemagglutinin (A/JAP/305/57, H2N2), induces CTLs that are directed to the MHC class Ib molecule, H2-M3. M3 preferably binds peptides bearing an N-terminal formylmethionine. In this study, we show that several related nonformylated peptides can induce anti-HA CTLs in vitro: MLIIW (the minimal epitope), derived from HA186-190 at the C-terminal end of HA173-190; MLIIWG; MLIIWGV; and MLIIWGI, as well as formylated MLIIW. The heptamer peptides correspond to a polymorphism of HA192 in H2 strains of influenza; they have the highest relative affinities for M3 of the nonformylated peptides and higher affinities than some formylated mitochondrial peptides. Depending on the affinity of the peptide, a range of concentrations can be used to induce CTLs. One nanomolar of the high affinity f-MLIIW peptide can induce anti-HA CTLs, whereas 100-fold more of the lower affinity MLIIW peptide is needed. Lines induced with high concentrations (1 microM or greater) of f-MLIIW recognize Ag poorly, and the most efficient CTLs are induced with the lowest concentrations of peptide. Analysis with a panel of anti-TCRVbeta Abs shows that different T cells respond to high vs low peptide; the repertoire of cells responding to higher concentrations is more diverse, consistent with the expansion of more, but less efficient, clones. Thus, peptide affinity and concentration should be considered together for generating efficient antipeptide CTLs in vitro. PMID- 10477566 TI - Human dendritic cells very efficiently present a heterologous antigen expressed on the surface of recombinant gram-positive bacteria to CD4+ T lymphocytes. AB - Recombinant Streptococcus gordonii expressing on the surface the C-fragment of tetanus toxin was tested as an Ag delivery system for human monocyte-derived dendritic cells (DCs). DCs incubated with recombinant S. gordonii were much more efficient than DCs pulsed with soluble C-fragment of tetanus toxin at stimulating specific CD4+ T cells as determined by cell proliferation and IFN-gamma release. Compared with DCs treated with soluble Ag, DCs fed with recombinant bacteria required 102- to 103-fold less Ag and were at least 102 times more effective on a per-cell basis for activating specific T cells. S. gordonii was internalized in DCs by conventional phagocytosis, and cytochalasin D inhibited presentation of bacteria-associated Ag, but not of soluble Ag, suggesting that phagocytosis was required for proper delivery of recombinant Ag. Bacteria were also very potent inducers of DC maturation, although they enhanced the capacity of DCs to activate specific CD4+ T cells at concentrations that did not stimulate DC maturation. In particular, S. gordonii dose-dependently up-regulated expression of membrane molecules (MHC I and II, CD80, CD86, CD54, CD40, CD83) and reduced both phagocytic and endocytic activities. Furthermore, bacteria promoted in a dose dependent manner DC release of cytokines (IL-6, TNF-alpha, IL-1beta, IL-12, TGF beta, and IL-10) and of the chemokines IL-8, RANTES, IFN-gamma-inducible protein 10, and monokine induced by IFN-gamma. Thus, recombinant Gram-positive bacteria appear a powerful tool for vaccine design due to their extremely high capacity to deliver Ags into DCs, as well as induce DC maturation and secretion of T cell chemoattractans. PMID- 10477567 TI - Characterization of RANTES- and aminooxypentane-RANTES-triggered desensitization signals reveals differences in recruitment of the G protein-coupled receptor complex. AB - The trafficking of lymphocyte populations is a complex process controlled by a vast array of molecules. In this process, cells must be able to sense small changes in chemoattractant gradients. Migration through a chemotactic gradient probably employs an on-off mechanism in which chemokine receptor desensitization, internalization, and recycling may be important steps. This multistep process requires the coordinated action of many factors, including G protein-coupled receptor kinases, arrestins, clathrin, and GTP-hydrolyzing proteins such as dynamin. In this report, we show that RANTES and its derivative, aminooxypentane (AOP)-RANTES, a potent RANTES antagonist as well as an inhibitor of HIV-1 infection, both promote CCR5 desensitization involving G protein-coupled receptor kinases-2 and beta-arrestin equally well. An important difference between the two molecules is that (AOP)-RANTES is more efficient than RANTES in promoting Ser/Thr phosphorylation of the receptor and association of G protein-coupled receptor kinases-2, beta-arrestin, and clathrin to the CCR5. After stimulation with either ligand, we observe rapid, transient association of dynamin to CCR5, implicating this protein in receptor sensitization, but this association is faster and longer lasting following (AOP)-RANTES stimulation. In summary, we show that chemokine receptor internalization takes place through the formation of clathrin vesicles and involves dynamin activity. We provide compelling evidence that the differences between RANTES and (AOP)-RANTES in G alpha i activation condition subsequent signaling events, including internalization and receptor recycling. PMID- 10477568 TI - Beta-glucan, a "specific" biologic response modifier that uses antibodies to target tumors for cytotoxic recognition by leukocyte complement receptor type 3 (CD11b/CD18). AB - beta-Glucans were identified 36 years ago as a biologic response modifier that stimulated tumor rejection. In vitro studies have shown that beta-glucans bind to a lectin domain within complement receptor type 3 (CR3; known also as Mac-1, CD11b/CD18, or alphaMbeta2-integrin, that functions as an adhesion molecule and a receptor for factor I-cleaved C3b, i.e., iC3b) resulting in the priming of this iC3b receptor for cytotoxicity of iC3b-opsonized target cells. This investigation explored mechanisms of tumor therapy with soluble beta-glucan in mice. Normal mouse sera were shown to contain low levels of Abs reactive with syngeneic or allogeneic tumor lines that activated complement, depositing C3 onto tumors. Implanted tumors became coated with IgM, IgG, and C3, and the absent C3 deposition on tumors in SCID mice was reconstituted with IgM or IgG isolated from normal sera. Therapy of mice with glucan- or mannan-rich soluble polysaccharides exhibiting high affinity for CR3 caused a 57-90% reduction in tumor weight. In young mice with lower levels of tumor-reactive Abs, the effectiveness of beta glucan was enhanced by administration of a tumor-specific mAb, and in SCID mice, an absent response to beta-glucan was reconstituted with normal IgM or IgG. The requirement for C3 on tumors and CR3 on leukocytes was highlighted by therapy failures in C3- or CR3-deficient mice. Thus, the tumoricidal function of CR3 binding polysaccharides such as beta-glucan in vivo is defined by natural and elicited Abs that direct iC3b deposition onto neoplastic cells, making them targets for circulating leukocytes bearing polysaccharide-primed CR3. Therapy fails when tumors lack iC3b, but can be restored by tumor-specific Abs that deposit iC3b onto the tumors. PMID- 10477569 TI - Effector CD4 T cells are biochemically distinct from the memory subset: evidence for long-term persistence of effectors in vivo. AB - Memory T cell responses are believed to be mediated by long-lived memory T cells that arise directly from a subset of short-lived, activated effector T cells that have reverted to the resting state. Although widely accepted, definitive proof that memory T cells arise from effectors is lacking because of the inability to reliably distinguish these subsets based on known phenotypic or functional parameters. We have used a biochemical approach to distinguish effector and memory CD4 T cell subsets and follow the differentiative fate of effector cells in vivo. When examined biochemically, effector and memory CD4 T cells are strikingly distinct and exhibit qualitative and quantitative differences in tyrosine phosphorylation. These effector-specific patterns were identical in effectors derived either from naive CD4 T cells (primary effectors) or memory CD4 T cells (memory effectors). To monitor the fate of effector cells in vivo, Ag activated CD4+ TCR-transgenic T cells were transferred into irradiated BALB/c mice. These TCR-transgenic CD4 T cells persisted in adoptive hosts for several months, gave a recall response to Ag, yet exhibited effector-specific biochemical profiles. These results suggest that a subset of effector CD4 T cells can persist in vivo and contribute to long-term immunity by mediating secondary immune responses. PMID- 10477570 TI - Migration of antigen-presenting B cells from peripheral to mucosal lymphoid tissues may induce intestinal antigen-specific IgA following parenteral immunization. AB - Parenterally administered immunizations have long been used to induce protection from mucosal pathogens such as Bordetella pertussis and influenza virus. We previously found that i.m. inoculation of mice with the intestinal pathogen, rotavirus, induced virus-specific Ab production by intestinal lymphocytes. We have now used adoptive transfer studies to identify the cell types responsible for the generation of virus-specific Ab production by gut-associated lymphoid tissue (GALT) after i.m. immunization. Three days after i.m. immunization with rotavirus, cells obtained from the draining peripheral lymph nodes of donor mice were transferred into naive recipient mice. We found that intestinal lymphocytes produced rotavirus-specific Igs (IgM, IgA, and IgG) 2 wk after transfer of either unfractionated cells, or unfractionated cells rendered incapable of cellular division by mitomycin C treatment. Additional studies demonstrated that rotavirus specific IgA, but not IgG, was produced by intestinal lymphocytes after transfer of purified B cells. Ig allotype analysis revealed that rotavirus-specific IgA was produced by intestinal B cells of recipient origin, suggesting that migration of Ag-presenting B cells from peripheral lymphoid tissues to GALT may contribute to the generation of mucosal IgA responses after parenteral immunization. Strategies that promote Ag uptake and presentation by B cells may enhance mucosal IgA production following parenteral immunization. PMID- 10477571 TI - Vasoactive intestinal peptide synergizes with TNF-alpha in inducing human dendritic cell maturation. AB - We investigated the effects of different neuropeptides on human dendritic cells (DC) maturation. Immature DC, derived from monocytes cultured for 6 days with IL 4 plus GM-CSF, have been exposed to somatostatin, substance P, or vasoactive intestinal peptide (VIP). Among these neuropeptides, only VIP induces the production of bioactive IL-12 and the neoexpression of CD83 on a fraction of the DC population, with an effect significant at 100 and 10 nM, respectively. These effects of VIP are dose-dependent, unaffected by polymixin B, and partly prevented by a VIP receptor antagonist. Although the effects of VIP alone remain modest, it synergizes with TNF-alpha to induce DC maturation. In the presence of a suboptimal concentration of TNF-alpha, which has no detectable effect on DC by itself, VIP induces the production of high levels of bioactive IL-12, the neoexpression of CD83 on almost all the DC population (with an effect significant at 10 and 0.1 nM, respectively), and the up-regulation of various adhesion and costimulatory molecule expression. Moreover, DC exposed to VIP plus a suboptimal concentration of TNF-alpha are as potent as mature DC obtained by treatment with an optimal concentration of TNF-alpha in stimulating allogenic T cell proliferation. Our data suggest that, in inflammatory sites, VIP may cooperate with proinflammatory mediators, such as TNF-alpha, to induce DC maturation. PMID- 10477572 TI - IL-4-producing gamma delta T cells that express a very restricted TCR repertoire are preferentially localized in liver and spleen. AB - IL-4-producing gamma delta thymocytes in normal mice belong to a distinct subset of gamma delta T cells characterized by low expression of Thy-1. This gamma delta thymocyte subset shares a number of phenotypic and functional properties with the NK T cell population. Thy-1dull gamma delta thymocytes in DBA/2 mice express a restricted repertoire of TCRs that are composed of the V gamma 1 gene product mainly associated with the V delta 6.4 chain and exhibit limited junctional sequence diversity. Using mice transgenic for a rearranged V gamma 1J gamma 4C gamma 4 chain and a novel mAb (9D3) specific for the V delta 6.3 and V delta 6.4 murine TCR delta chains, we have analyzed the peripheral localization and functional properties of gamma delta T cells displaying a similarly restricted TCR repertoire. In transgenic mice, IL-4 production by peripheral gamma delta T cells was confined to the gamma delta+9D3+ subset, which contains cells with a TCR repertoire similar to that found in Thy-1dull gamma delta thymocytes. In normal DBA/2 mice such cells represent close to half of the gamma delta T cells present in the liver and around 20% of the splenic gamma delta T cells. PMID- 10477573 TI - Identification of a new type of invariant V alpha 14+ T cells and responsiveness to a superantigen, Yersinia pseudotuberculosis-derived mitogen. AB - We examined the expression of the H4 T cell activation marker in thymic T cell subpopulations and found that TCR-alpha beta+ CD4+ thymic T cells are segregated into three subpopulations based upon H4 levels. Thymic T cells with either no or low H4 expression differentiate via the mainstream differentiation pathway in the thymus. H4int thymic T cells, which express a skewed V beta repertoire of V beta 2, -7, and -8 in their TCRs, show the phenotype of NKT cells: CD44high, Ly6Chigh, NK1.1+, and TCR-alpha beta low. H4high thymic T cells also show a skewed V beta repertoire, V beta 2, -7, and -8, and predominantly express an invariant V alpha 14-J alpha 281+ alpha-chain in their TCRs but constitute a distinct population in that they are CD44int, Ly6C-, NK1.1-, and TCR-alpha beta high. Thus, invariant V alpha 14+ thymic T cells consist of ordinary NKT cells and a new type of T cell population. V beta 7+ and V beta 8.1+ invariant V alpha 14+ thymic T cells are present in DBA/2 mice, which carry mammary tumor virus-7-encoded superantigens, in comparable levels to those in BALB/c mice. Furthermore, V beta 7+ invariant V alpha 14+ thymic T cells in DBA/2 mice are in the immunologically responsive state, and Yersinia pseudotuberculosis-derived mitogen-induced V beta 7+ invariant V alpha 14+ thymic T cell blasts from DBA/2 and BALB/c mice exhibited equally enhanced responses upon restimulation with Y. pseudotuberculosis-derived mitogen. Thus, invariant V alpha 14+ thymic T cells that escape negative selection in DBA/2 mice contain T cells as functionally mature as those in BALB/c mice. PMID- 10477574 TI - A novel apoptotic pathway in quiescent lymphocytes identified by inhibition of a post-proline cleaving aminodipeptidase: a candidate target protease, quiescent cell proline dipeptidase. AB - The vast majority of lymphocytes in vivo persist in a quiescent state. These resting lymphocytes are maintained through a cellular program that suppresses apoptosis. We show here that quiescent PBMC, but not activated PBMC or transformed lymphocytes, die in the presence of highly specific post-proline aminodipeptidase inhibitors. This form of death has the hallmarks of apoptosis, such as phosphatidylserine externalization and loss of mitochondrial transmembrane potential. However, it differs from apoptosis induced by gamma irradiation in the same cells or by Fas ligation in transformed lymphocytes in terms of caspase involvement. In addition, the aminodipeptidase inhibitor-induced cell death, but not gamma-irradiation-mediated apoptosis, can be prevented by inhibition of the proteasome complex. The target of these inhibitors is not CD26/DPPIV, but probably a novel serine protease, quiescent cell proline dipeptidase, that we have recently isolated and cloned. These studies will yield a better understanding of the requirements and the mechanisms that mediate quiescent lymphocyte homeostasis in vivo. PMID- 10477575 TI - IL-12 acts selectively on CD8 alpha- dendritic cells to enhance presentation of a tumor peptide in vivo. AB - Previous work has shown that a significant proportion of murine splenic dendritic cells (DC) express a high affinity receptor for IL-12, thus accounting for the adjuvanticity of the cytokine when DBA/2 mice are transferred with syngeneic DC exposed in vitro to rIL-12 and an otherwise poorly immunogenic tumor peptide. In DBA/2 mice, splenic DC consist of 90-95% CD8- and 5-10% CD8+ cells. To detect any difference in IL-12 responsiveness among phenotypically distinct DC subtypes, enriched CD8- (>99% pure) and CD8+ ( approximately 80% pure) populations of DC from DBA/2 spleens were assayed for APC function in vivo following exposure to rIL-12 and tumor peptide in vitro. Unlike unfractionated DC, the CD8- fraction was capable of effective presentation of the peptide even when the cells had not been pretreated with IL-12 before peptide pulsing. The addition of as few as 3% CD8+ cells during pulsing blocked in vivo priming by the CD8- fraction. However, pretreatment of CD8- DC with IL-12 before cell mixing and peptide pulsing ablated the inhibitory effect of the CD8+ fraction. CD8-, but not CD8+, DC showed significant message expression for the beta 1 and beta 2 subunits of the IL-12 receptor. These data suggest that a minority population of CD8+ DC, which appeared to secrete IL-10 in vitro, negatively regulates the induction of T cell reactivity by peptide-loaded CD8- DC in DBA/2 mice. However, the CD8- fraction can be primed by IL-12 to overcome the inhibitory effect of the CD8+ subtype. PMID- 10477576 TI - TNF-alpha-induced growth suppression of CD34+ myeloid leukemic cell lines signals through TNF receptor type I and is associated with NF-kappa B activation. AB - Conflicting results have been reported regarding the effect of TNF-alpha on the growth of human primitive hemopoietic cells. In this study, we have examined the effect of TNF-alpha on the proliferation of several CD34+/CD38+ (KG-1, TF-1) and CD34+/CD38- (KG-1a, TF-1a) myeloid leukemic progenitor cell lines. Our data show that TNF-alpha markedly inhibits the growth of these cells in both liquid and soft agar cultures. Addition of GM-CSF or IL-3 does not prevent TNF-alpha-induced growth inhibition. Flow cytometry analyses of propidium iodide-stained cells demonstrated cell death of all four cell lines, as judged by the presence of cells with hypodiploid DNA content after exposure of cells to TNF-alpha for 4 days. Annexin V assays detected apoptosis in TF-1, but not in TF-1a, KG-1, and KG 1a cells in terms of translocation of phosphatidylserine shortly after TNF-alpha treatment. Neutralizing anti-TNF receptor type I (TNFR-I; p55) Ab almost completely reversed TNF-alpha-induced growth inhibition in both liquid and soft agar cultures, whereas anti-TNFR-II (p75) Ab had only a marginal effect. TNF alpha rapidly induced marked activation of nuclear transcription factor NF-kappa B in all 4 cell lines. The majority of this effect was abolished by the type I receptor Ab, whereas the type II receptor neutralizing Ab had no effect. Our data also show that TNF-alpha is incapable of inducing activation of the mitogen activated protein kinase pathway in these leukemic cell lines. PMID- 10477577 TI - Human rheumatoid factor production is dependent on CD40 signaling and autoantigen. AB - High-affinity pathologic rheumatoid factor (RF) B cells occur in autoimmune diseases such as rheumatoid arthritis, but are deleted in healthy individuals. The reasons for the survival and differentiation of these autoreactive B cells in rheumatoid arthritis are not known. Previous studies in mice transgenic for a human IgM RF have shown that peripheral encounter with soluble human IgG leads to deletion of high-affinity RF B cells; however, deletion can be prevented when concomitant T cell help is provided. This study aimed to further discern the minimal factors necessary not only for the in vivo survival of RF B cells, but also for their differentiation into Ab-secreting cells. The combination of MHC class II-reactive T cells and Ag induced the production of RF in human IgM RF transgenic mice, while either stimulus alone was ineffective. Neutralizing Abs against CD40 ligand (CD40L), but not against IL-4 or IL-15, abrogated IgM-RF production. Moreover, blockade of CD40L-CD40 allowed IgG to delete the RF precursor cells. Most importantly, activating Abs to CD40 could substitute entirely for T cell help in promoting the survival of RF precursors and in stimulating RF synthesis in T cell deficient animals. The data indicate that CD40 signaling alone can prevent deletion of RF B cells by Ag and in the presence of IgG is sufficient to trigger RF synthesis. The results suggest that selective induction of apoptosis in high-affinity RF B cells may be achieved by blockade of CD40L-CD40 interaction. PMID- 10477578 TI - Reciprocal desensitization of CCR5 and CD4 is mediated by IL-16 and macrophage inflammatory protein-1 beta, respectively. AB - The ability of HIV-1 gp120 to inhibit chemokine signaling prompted us to determine whether signaling through CD4 by a natural ligand, IL-16, could alter cellular responsiveness to chemokine stimulation. These studies demonstrate that IL-16/CD4 signaling in T lymphocytes results in a selective loss of macrophage inflammatory protein (MIP)-1 beta/CCR5-induced chemotaxis. There was no effect on monocyte chemoattractant protein-2/CCR1, -2, or -3-induced chemotaxis. Desensitization of CCR5 by IL-16 required at least 10 min of pretreatment; no modulation of CCR5 expression was observed, nor was MIP-1 beta binding to CCR5 altered. Using murine T cell hybridomas transfected to express native or mutated forms of CD4, it was determined that IL-16/CD4 induces a p56lck-dependent signal that results in desensitization of CCR5. The desensitization process is reciprocal and again selective, as prior CCR5 stimulation, but not CCR1, -2, or 3 stimulation, completely inhibits IL-16/CD4-induced T cell migration. Of interest, while p56lck enzymatic activity is not required for IL-16-induced migration, it was required for desensitization of CCR5. These studies indicate the existence of reciprocal receptor cross-desensitization between CD4 and CCR5 induced by two proinflammatory cytokines and suggest a selective relationship between the two receptors. PMID- 10477579 TI - The role of the common cytokine receptor gamma-chain in regulating IL-2 dependent, activation-induced CD8+ T cell death. AB - IL-2-dependent, activation-induced T cell death (AICD) plays an important role in peripheral tolerance. Using CD8+ TCR-transgenic lymphocytes (2C), we investigated the mechanisms by which IL-2 prepares CD8+ T cells for AICD. We found that both Fas and TNFR death pathways mediate the AICD of 2C cells. Neutralizing IL-2, IL 2R alpha, or IL-2R beta inhibited AICD. In contrast, blocking the common cytokine receptor gamma-chain (gamma c) prevented Bcl-2 induction and augmented AICD. IL-2 up-regulated Fas ligand (FasL) and down-regulated gamma c expression on activated 2C cells in vitro and in vivo. Adult IL-2 gene-knockout mice displayed exaggerated gamma c expression on their CD8+, but not on their CD4+, T cells. IL 4, IL-7, and IL-15, which do not promote AICD, did not influence FasL or gamma c expression. These data provide evidence that IL-2 prepares CD8+ T lymphocytes for AICD by at least two mechanisms: 1) by up-regulating a pro-apoptotic molecule, FasL, and 2) by down-regulating a survival molecule, gamma c. PMID- 10477581 TI - SLP-76 binding to p56lck: a role for SLP-76 in CD4-induced desensitization of the TCR/CD3 signaling complex. AB - Nonreceptor protein tyrosine kinases and associated substrates play a pivotal role in Ag receptor stimulation of resting cells and in the initiation of activation-induced cell death (AICD) of preactivated T cells. CD4-associated p56lck has been implicated not only in the activation of primary T cells, but also in the inhibition of T cell responses. We have previously shown that CD4+ T cell clones can be rescued from AICD when surface CD4 is engaged before the TCR stimulus. In this study, we show that prevention of AICD is associated with a CD4 dependent inhibition of TCR-triggered tyrosine phosphorylation of the Src homology 2 domain-containing leukocyte protein of 76 kDa (SLP-76) and Vav. We provide evidence for a SLP-76 interaction with Src homology 3 domains of p56lck and identify amino acids 185-194 of SLP-76 as relevant docking site. In view of the multiple functions of p56lck and SLP-76/Vav in the initiation of TCR/CD3/CD4 signaling, we propose a model for the CD4-dependent inhibition of TCR signaling and AICD of preactivated T cells. Our data suggest that preformed activation complexes of adapter proteins and enzymes in the vicinity of the CD4/p56lck complex are no longer available for the TCR signal when CD4 receptors are engaged before TCR stimulation. PMID- 10477580 TI - Differential control of autoantibodies and lymphoproliferation by Fas ligand expression on CD4+ and CD8+ T cells in vivo. AB - We have previously shown that the gld autoimmune syndrome is suppressed in lethally irradiated gld mice reconstituted with a mixture of normal and gld bone marrow (BM). Furthermore, in vivo depletion of normal Thy-1+ cells restores lymphoproliferation and autoantibody production in such chimeras, suggesting that T cells bearing Fas ligand are responsible for correcting the gld defect. In this study, mixed-BM chimeras lacking either normal CD4+ (B6CD4KO-B6gld) or normal CD8+ T cells (B6CD8KO-B6gld) were generated to determine the contribution of the normal T cell subsets to disease suppression. Lymphoproliferation was completely suppressed in B6CD4KO-B6gld chimeras but only modestly in B6CD8KO-B6gld chimeras. On the other hand, both types of mixed-BM chimeras had incomplete effects on the suppression of serum autoantibodies when compared with B6gld reconstituted with isologous BM. These results suggest that both T cell subsets provide Fas ligand to suppress immune cells responsible for autoantibody production; however, CD8+ T cells are mainly responsible for preventing lymphoproliferation. PMID- 10477582 TI - Role of the costimulatory molecule CD28 in the development of lupus in MRL/lpr mice. AB - MRL/Mpj-lpr/lpr (MRL/lpr) mice develop autoimmune disorders, including lymphoproliferation, glomerulonephritis, autoantibody production, and hypergammaglobulinemia. To investigate the role of the costimulatory molecule CD28 in the development of these disorders, MRL/lpr mice lacking CD28 were generated by gene targeting. Compared with CD28+/+ MRL/lpr mice, CD28-/- MRL/lpr mice showed decreased lymphadenopathy but increased splenomegaly associated with the expansion of abnormal B220+ TCRalphabeta+ T cells. Although levels of IgM Abs were unchanged in CD28-/- MRL/lpr mice, the production of anti-DNA IgG Abs and IgG rheumatoid factors were suppressed. IgG deposition in the glomeruli was markedly decreased, and the development of glomerulonephritis was significantly retarded. Furthermore, renal vasculitis and arthritis were absent in CD28-/- MRL/lpr mice. These results indicate that, although CD28 is not required for the generation of the abnormal T cell population in MRL/lpr mice, it does play an important role in the development of autoimmune disease in these animals. PMID- 10477583 TI - B cell antigen receptor-mediated activation of cyclin-dependent retinoblastoma protein kinases and inhibition by co-cross-linking with Fc gamma receptors. AB - Cross-linking the B cell Ag receptor (BCR) to surface Fc receptors for IgG (Fc gamma R) inhibits G1-to-S progression; the mechanism by which this occurs is not completely known. We investigated the regulation of three key cell cycle regulatory components by BCR-Fc gamma R co-cross-linking: G1-cyclins, cyclin dependent kinases (Cdks), and the retinoblastoma gene product (Rb). Rb functions to suppress G1-to-S progression in mammalian cells. Rb undergoes cell-cycle dependent phosphorylation, leading to its inactivation and thereby promoting S phase entry. We demonstrate in this paper for the first time that BCR-induced Rb phosphorylation is abrogated by co-cross-linking with Fc gamma R. The activation of Cdk4/6- and Cdk2-dependent Rb protein kinases is concomitantly blocked. Fc gamma R-mediated inhibition of Cdk2 activity results in part from an apparent failure to express Cdk2 protein. By contrast, inhibition of Cdk4/6 activities is not due to suppression of Cdk4/6 or cyclins D2/D3 expression or inhibition of Cdk activating kinase activity. Cdk4- and Cdk6-immune complexes recovered from B cells following BCR-Fc gamma R co-cross-linking are devoid of coprecipitated D type cyclins, indicating that inhibition of their Rb protein kinase activities is due in part to the absence of bound D-type cyclin. Thus, BCR-derived activation signals that up-regulate D-type cyclin and Cdk4/6 protein expression remain intact; however, Fc gamma R-mediated signals block cyclin D-Cdk4/6 assembly or stabilization. These results suggest that assembly or stabilization of D-type cyclin holoenzyme complexes 1) is an important step in the activation of Cdk4/6 by BCR signals, and 2) suffice in providing a mechanism to account for inhibition of BCR-stimulated Rb protein phosphorylation by Fc gamma R. PMID- 10477584 TI - Recombinant IFN-alpha (2b) increases the expression of apoptosis receptor CD95 and chemokine receptors CCR1 and CCR3 in monocytoid cells. AB - IFN-alpha-2b, known as potent immune modulator, can either inhibit or enhance immune cell activity within the tightly regulated microenvironment of inflammation, depending upon the concentration of the cytokine and the activation stage of the cell. Chemokine receptors, which not only mediate chemotaxis of immune cells to the site of inflammation but also affect cellular activation by transferring corresponding signals, represent yet another level of immune regulation. Here we demonstrate that IFN-alpha increases the expression of CCR1 and CCR3 in primary mononuclear phagocytes, as well as in the monocytoid cell line U937. Enhanced receptor mRNA expression correlated with functional readouts such as increased intracellular calcium mobilization and cell migration in response to ligands. Expression of CCR2b, CCR4, CCR5, and CXCR4 was unchanged or decreased after IFN-alpha treatment. These observations indicate a differentially regulated cellular signaling relationship of IFN-alpha pathways and chemokine receptor expression. We also provide evidence that, under these conditions, IFN alpha treatment increased the expression of CD95 (Fas, Apo1), resulting in enhanced susceptibility to apoptosis. Taken together, these data add important information for the rational application of IFN-alpha (2b) in immune and cancer therapies. PMID- 10477585 TI - Stochastic acquisition of Qa1 receptors during the development of fetal NK cells in vitro accounts in part but not in whole for the ability of these cells to distinguish between class I-sufficient and class I-deficient targets. AB - Fetal mouse NK cells are grossly deficient in the expression of Ly49 molecules yet show a limited ability to distinguish between wild-type and MHC class I deficient target cells. In this paper we report that during their development in vitro from immature thymic progenitors, a proportion of C57BL/6 fetal NK cells acquires receptors for a soluble form of the nonclassical class I molecule Qa1b associated with the Qdm peptide, but not for soluble forms of the classical class I molecules Kb and Db. The acquisition of these Qa1 receptors occurs in a stochastic manner that is strictly controlled by cytokines, and in particular is strongly inhibited by IL-4. All fetal NK clones tested, including those that lack detectable Qa1 receptors, express mRNA for CD94 and for both inhibitory and noninhibitory members of the NKG2 family. Fetal NK cells lacking receptors for Qa1 (and also for classical class I molecules) cannot distinguish between wild type and class I-deficient blasts but, surprisingly, distinguish efficiently between certain wild-type and class I-deficient tumor cells. A variant line that lacks several members of the NKG2 family kills both types of tumor cell equally well, suggesting the existence of NKG2-containing inhibitory receptors that recognize as yet undefined nonclassical class I molecules of restricted distribution. PMID- 10477586 TI - The human B cell response to IL-13 is dependent on cellular phenotype as well as mode of activation. AB - Normal mature quiescent human B lymphocytes, isolated as a function of buoyant density, require activation for up-regulation of IL-13R constituents. Cell activation through a combination of surface Ig and CD40 receptor ligation leads to the most substantial message production for IL-13Ralpha1. Functional consequences of this receptor variation, in initially quiescent cells, includes demonstrable effects on cellular proliferation in response to ligand exposure. Variations in the method of surface activation, with particular emphasis on the CD40 receptor, reveals that immobilized CD40 ligand may be sufficient, in and of itself, to up-regulate IL-13Ralpha1, which may bear significance for B-lymphocyte bystander proliferation. Regulation of the IL-13Ralpha1 protein and message also differs as a function of cellular phenotype. Although values are greater in memory than naive B cells, as they are initially isolated from extirpated tonsils, variations in the magnitude of message and protein, as a function of surface stimulation, are more substantial in the naive subset. The magnitude of variation in message production in naive cells is associated with a more vigorous proliferative response to IL-13 than seen in memory lymphocytes. The cellular response to IL-13, as a function of activation and phenotype, is the converse of that demonstrated for IL-2. Evaluation of proliferation, receptor message, ligand binding protein production, and the response to putatively synergistic cytokines reveals that IL-2 is the predominant lymphokine utilized by memory cells. This is in contradistinction to IL-13, which along with IL-4, are the predominant moieties for naive lymphocytes. PMID- 10477587 TI - CD40-CD40 ligand costimulation is required for generating antiviral CD4 T cell responses but is dispensable for CD8 T cell responses. AB - This study documents a striking dichotomy between CD4 and CD8 T cells in terms of their requirements for CD40-CD40 ligand (CD40L) costimulation. CD40L-deficient ( /-) mice made potent virus-specific CD8 T cell responses to dominant as well as subdominant epitopes following infection with lymphocytic choriomeningitis virus. In contrast, in the very same mice, virus-specific CD4 T cell responses were severely compromised. There were 10-fold fewer virus-specific CD4 T cells in CD40L-/- mice compared with those in CD40L+/+ mice, and this inhibition was seen for both Th1 (IFN-gamma, IL-2) and Th2 (IL-4) responses. An in vivo functional consequence of this Th cell defect was the inability of CD40L-/- mice to control a chronic lymphocytic choriomeningitis virus infection. This study highlights the importance of CD40-CD40L interactions in generating virus-specific CD4 T cell responses and in resolving chronic viral infection. PMID- 10477588 TI - Selective retention of activated CD8+ T cells by the normal liver. AB - Activation-induced cell death resulting in peripheral deletion of CD8+ T cells is associated with the accumulation of large numbers of apoptotic T cells in the liver. The hypothesis that this accumulation results from the intrahepatic trapping of T cells from the circulating pool predicts that the liver should retain T cells, which subsequently undergo apoptosis. Here we test this prediction. Perfusion of the liver with lymphocyte mixtures showed retention of activated, but neither resting nor apoptosing, T cells. This trapping was selective for CD8+ cells and was mediated primarily by ICAM-1 constitutively expressed on sinusoidal endothelial cells and Kupffer cells. T cells trapped in the liver became apoptotic. The normal liver is therefore a "sink" for activated T cells. PMID- 10477589 TI - Roles of integrins and CD44 on the adhesion and migration of fetal liver cells to the fetal thymus. AB - Adhesion and migration of mouse fetal liver (FL) cells to the thymus were investigated using cells from green fluorescent protein transgenic (GFP+) mice. FL cells from GFP+ embryos at 12 gestational days (E12) of mice were incubated with 2'-deoxyguanosine-treated fetal thymus lobe (from E14) by thymic repopulation (hanging drop) culture methods. GFP+ cells were observed in the thymus lobe at the end of the repopulation culture period. A large part of the infiltrated cells expressed CD44 until day 2 of culture on a permeable membrane, then lost the expression. CD25 expression was observed from day 1 to day 4. Around day 8, GFP+ cells became both CD4+ and CD8+. The results support the early observation of the sequential expression of CD44, CD25, and CD4/8 during the early stages of thymocyte development. When anti-CD44 mAb was added at the beginning of the repopulation culture period, GFP+ FL cells adhered to the surface of the thymus lobe but did not migrate into the thymus. Pretreatment of the thymus with hyaluronidase or hyaluronate produced results similar to the results of anti-CD44 treatment. On the other hand, the addition of anti-integrin alpha4 mAb inhibited adhesion to the thymus, and almost no GFP+ cells were seen on the surface of the thymus lobe. The data suggest that integrin alpha4 and CD44 play different roles, i.e., integrin alpha4 is required for the adhesion of FL cells to the thymus lobe and CD44 is required for the migration of the cells into the thymus. PMID- 10477590 TI - Alloreactive and syngeneic CTL are comparably dependent on interaction with MHC class I alpha-helical residues. AB - The molecular basis for the difference in the strength of T cell responses to self vs alloantigens is unknown, but may reflect how T cells are selected in the thymus. Because T cells with a high affinity for foreign as opposed to self MHC molecules are able to mature, it has been proposed that alloreactive T cells may be more strongly dependent upon interaction with MHC residues than are self restricted T cells. This study was undertaken to rigorously address this hypothesis. Whereas other studies have compared self vs alloantigen recognition of different MHC alleles by a single T cell clone, we have compared self vs alloantigen recognition of a single MHC allele, H-2Ld, by a large panel of self restricted and alloreactive T cell clones. Target cells expressing Ld molecules mutated at several different potential TCR contact residues were analyzed to determine which residues are important for recognition by self-restricted vs alloreactive T cells. We unequivocally demonstrate that self-restricted and alloreactive T cells do not differ, but rather are comparably dependent on interaction with MHC residues. Importantly, both self-restricted and alloreactive T cells are dependent upon the same MHC residues as primary contacts and, in addition, share a common recognition pattern of Ld. Furthermore, our analysis enables us to provide a model for allotype-specific T cell recognition of Ld vs Kb class I molecules. PMID- 10477591 TI - In vivo immunomodulation following intradermal injection with DNA encoding IL-18. AB - IL-18, a recently identified cytokine synthesized by different cell types, including Kupffer cells, activated macrophages, and keratinocytes, induces IFN gamma production by T cells and NK cells. The cDNA encoding IL-18 with its natural signal peptide was cloned under control of the CMV promoter and injected into the skin of mice. A single intradermal injection of this construction led to efficient in vivo expression of IL-18 in cutaneous dermal cells and induced IFN gamma mRNA production, indicating that it was produced in a biologically active form. In addition, a massive cellular infiltrate was observed in the skin 2 days after injection. When the mice were subsequently infected with Mycobacterium bovis bacillus Calmette-Guerin (BCG), they produced lower levels of anti-BCG Abs than control animals. However, in contrast to their lowered humoral immune response, the mice produced higher amounts of Ag-specific IFN-gamma after in vitro restimulation, as compared with the controls. Therefore, injection of DNA encoding IL-18 into the skin modulates both Ag-specific humoral and T cell responses upon mycobacterial infection. It increases the Th1 type response, which may be particularly useful for the development of new immunotherapeutic or immunoprotective approaches against infections by intracellular parasites, such as mycobacteria. PMID- 10477592 TI - A role for IL-16 in the cross-talk between dendritic cells and T cells. AB - Dendritic cells (DCs) in the periphery capture and process Ags, migrate to lymphoid organs, and initiate immune responses in T cells. IL-16, the soluble ligand for CD4, is a potent chemoattractant for CD4+ T cells, eosinophils, and monocytes and is mainly derived from activated T cells. Because migration is a fundamental property of DCs, we asked whether IL-16 induces chemotaxis in DCs and whether DCs are a source of IL-16. DCs were generated by culture of monocytes in IL-4 and GM-CSF for 6 days and subsequently highly purified employing magnetic beads. Migration was assayed by nitrocellulose and polycarbonate filter-based assays, and distinction of chemotaxis and chemokinesis was performed by a checkerboard analysis. Messenger RNA and protein data revealed constitutive expression and release of IL-16 by day-6 DCs. Gradients of rIL-16 induced a chemotactic response of DCs. Furthermore, the chemotactic activity of DC supernatant toward DCs themselves and T cells was mainly due to IL-16, because the addition of neutralizing Abs completely abrogated the migratory response. However, after induction of maturation by the addition of TNF-alpha and PGE2 DCs, neither expressed IL-16 mRNA nor produced IL-16 protein. We conclude that IL-16 may play a role in the trafficking of DCs and may be a major chemotactic signal from DCs toward themselves and toward T cells. PMID- 10477593 TI - Human fibroblasts transduced with CD80 or CD86 efficiently trans-costimulate CD4+ and CD8+ T lymphocytes in HLA-restricted reactions: implications for immune augmentation cancer therapy and autoimmunity. AB - Augmenting immunogenicity by genetically modifying tumor cells to express costimulatory molecules has proven to be a promising therapeutic strategy in murine tumor models and is currently under investigation in human clinical trials for metastatic cancer. However, there are significant technical and logistic problems associated with implementing strategies requiring direct gene modification of primary tumor cells. In an effort to circumvent these problems, we are developing a strategy in which the costimulatory signal required for tumor specific T lymphocyte activation is provided by a genetically modified human fibroblast (trans-costimulation). We have evaluated the efficiency of CD80- and CD86-mediated trans-costimulation in the activation of human CD8+ and CD4+ T lymphocytes in MHC class I- and class II-restricted lymphoproliferation reactions. Our studies demonstrate that the efficiency of CD80- or CD86-mediated trans-costimulation of purified human CD8+ and CD4+ T lymphocytes is comparable to cis-costimulation under defined conditions. Moreover, a dose-response relationship consistent with the predicted two-hit kinetics of the reaction was evident in trans-costimulation reactions in which the ratio of target cells expressing either signal 1 or signal 2 was varied incrementally from 1:10 to 10:1. Importantly, the level of cell-surface CD86 required for trans costimulation is equivalent to that constitutively expressed by human peripheral blood monocytes. These results may have significant implications for the clinical implementation of this type of cancer immunotherapy and also raise questions about the possibility of trans-costimulating autoreactive T lymphocytes in vivo. PMID- 10477594 TI - Survival, maturation, and function of CD11c- and CD11c+ peripheral blood dendritic cells are differentially regulated by cytokines. AB - Two types of dendritic cells (DC) are circulating in human blood and can be identified by their differential expression of the myeloid Ag CD11c. In this study, we show that CD11c- peripheral blood (PB)-DC correspond to plasmacytoid DC of lymphoid tissue not only by their surface Ag expression profile but, more impressively, by their peculiar ultramorphology. We also demonstrate that CD11c- and CD11c+ DC differ in the quality of their response to and in their requirement for certain cytokines. Freshly isolated CD11c- cells depend on IL-3 for survival and use autocrine or exogenous TNF-alpha as maturation signal, leading to the appearance of a highly dendritic phenotype, the up-regulation and redistribution of MHC class II from lysosomal compartments to the plasma membrane, the increased expression of costimulatory molecules, and the switch from a high Ag-processing to a low Ag-processing/potent accessory cell mode. Surprisingly, IL-4 efficiently killed freshly isolated CD11c- PB-DC, but did not impair the viability of CD11c+ PB-DC and, together with GM-CSF, induced maturation of these cells. A direct functional comparison revealed that neo-Ag-modified and subsequently matured CD11c- but to a lesser extent CD11c+ DC were able to prime naive Ag-specific CD4+ T cells. Our findings show that two diverse DC types respond to certain T cell derived cytokines in a differential manner and, thus, suggest that suppression or activation of functionally diverse DC types may be a novel mechanism for the regulation of the quantity and quality of immune responses. PMID- 10477595 TI - Effect of vascular endothelial growth factor and FLT3 ligand on dendritic cell generation in vivo. AB - The cytokine FLT3 ligand (FL) enhances dendritic cell (DC) generation and has therefore been proposed as a means to boost antitumor immunity. Vascular endothelial growth factor (VEGF) is produced by a large percentage of tumors and is required for development of tumor neovasculature. We previously showed that VEGF decreases DC production and function in vivo. In this study, we tested the hypothesis that VEGF regulates FL effects on DC generation. In seven experiments, four groups of mice were treated with PBS, VEGF alone (100 ng/h), FL alone (10 microgram/day), or with the combination of FL and VEGF. VEGF and PBS were administered continuously for 14 days via s.c. pumps. FL was given s.c. daily for 9 days, beginning on day 4. Tissues were collected and the number, phenotype, and function of lymph node, splenic, and thymic DCs were analyzed on day 14. As expected, treatment with FL resulted in a marked increase in the number of lymph node and spleen DCs and a smaller increase in thymic DC. Pretreatment of mice with VEGF inhibited these FL effects in lymph nodes and thymus by about 50%, whereas spleen DC numbers were undiminished by VEGF. VEGF treatment in vivo also inhibited the ability of FL to increase the number of hemopoietic precursor cells and the level of maturity exhibited by DC derived from these hemopoietic precursor cells in vitro. VEGF inhibited FL-inducible activation of transcription factor NF-kappaB. These data suggest that VEGF interferes with the ability of FL to promote dendritic cell differentiation from bone marrow progenitor cells in mice and therefore may decrease the therapeutic efficacy of FL in settings where increased numbers of DCs might provide clinical benefits. PMID- 10477596 TI - Selective diapedesis of Th1 cells induced by endothelial cell RANTES. AB - Differentiated CD4 T cells can be divided into Th1 and Th2 types based on the cytokines they produce. Differential expression of chemokine receptors on either the Th1-type or the Th2-type cell suggests that Th1-type and Th2-type cells differ not only in cytokine production but also in their migratory capacity. Stimulation of endothelial cells with IFN-gamma selectively enhanced transmigration of Th1-type cells, but not Th2-type cells, in a transendothelial migration assay. Enhanced transmigration of Th1-type cells was dependent on the chemokine RANTES produced by endothelial cells, as indicated by the findings that Ab neutralizing RANTES, or Ab to its receptor CCR5, inhibited transmigration. Neutralizing Ab to chemokines macrophage-inflammatory protein-1alpha or monocyte chemotactic protein-1 did not inhibit Th1 selective migration. Whereas anti-CD18 and anti-CD54 blocked basal levels of Th1-type cell adherence to endothelial cells and also inhibited transmigration, anti-RANTES blocked only transmigration, indicating that RANTES appeared to induce transmigration of adherent T cells. RANTES seemed to promote diapedesis of adherent Th1-type cells by augmenting pseudopod formation in conjunction with actin rearrangement by a pathway that was sensitive to the phosphoinositol 3-kinase inhibitor wortmannin and to the Rho GTP binding protein inhibitor, epidermal cell differentiation inhibitor. Thus, enhancement of Th1-type selective migration appeared to be responsible for the diapedesis induced by interaction between CCR5 on Th1-type cells and RANTES produced by endothelial cells. Further evidence that CCR5 and RANTES play a modulatory role in Th1-type selective migration derives from the abrogation of this migration by anti-RANTES and anti-CCR5 Abs. PMID- 10477597 TI - TNF-mediated activation of the stress-activated protein kinase pathway: TNF receptor-associated factor 2 recruits and activates germinal center kinase related. AB - TNF-induced activation of stress activated protein kinases (SAPKs, Jun NH2 terminal kinases) requires TNF receptor associated factor 2 (TRAF2). TRAF2 is a potent activator of a 95-kDa serine/threonine kinase termed germinal center kinase related (GCKR, also referred to as KHS1), which signals activation of the SAPK pathway. Consistent with a role for GCKR in TNF- induced SAPK activation, a kinase-inactive mutant of GCKR is a dominant negative inhibitor of TRAF2-induced SAPK activation. Here we show that TRAF2 interacts with GCKR. This interaction depended upon the TRAF domain of TRAF2 and the C-terminal 150 aa of GCKR. The full activation of GCKR by TRAF2 required the TRAF2 RING finger domain. TNF treatment of a T cell line, Jurkat, increased both GCRK and SAPK activity and enhanced the coimmunoprecipitation of GCKR with TRAF2. Similar results were found with the B cell line HS-Sultan. These findings are consistent with a model whereby TNF signaling results in the recruitment and activation of GCKR by TRAF2, which leads to SAPK activation. PMID- 10477598 TI - The mode of ligand recognition by two peptide:MHC class I-specific monoclonal antibodies. AB - The Ig superfamily members TCR and B cell receptor (BCR) share high structural and amino acid homology, yet interact with Ags in a distinct manner. The overall shape of the TCR ligand is rather constant, with the variation coming from the MHC polymorphism and the peptide heterogeneity. Consequently, the TCR alpha- and beta-chains form a relatively flat ligand-binding site that interacts with the peptide:MHC (pep:MHC) ligand in a fixed diagonal orientation relative to the MHC alpha-helices, with the alpha- and beta-chains of the TCR contacting the N and C termini of the pep:MHC complex, respectively. By contrast, the shape of BCR ligands varies dramatically, and the BCR exhibits much greater variability of the Ag-binding site. The mAbs 25-D1.16 (D1) and 22-C5.9 (C5), specific for the OVA 8:H-2Kb complex, allowed us to directly compare how TCR and BCR approach the same ligand. To that effect, we mapped D1 and C5 footprints over the OVA-8:H-2Kb complex. Using peptide variants and mutant MHC molecules, we show that the D1 and C5 contacts with the OVA-8:Kb complex C terminus overlap with the TCR beta-chain footprint, but that this footprint also extends to the regions of the molecule not contacted by the TCR. These studies suggest that D1 and C5 exhibit a hybrid mode of pep:MHC recognition, in part similar to that of the TCR beta-chain and in part similar to the conventional anti-MHC Ab. PMID- 10477599 TI - An activation-responsive element in single C motif-1/lymphotactin promoter is a site of constitutive and inducible DNA-protein interactions involving nuclear factor of activated T cell. AB - Single C motif-1 (SCM-1)/lymphotactin is a C-type chemokine whose expression is activation dependent, cyclosporin A sensitive and restricted to CD8+ T cells, double-negative thymocytes, gammadelta-type T cells, and NK cells. In humans, there are two highly homologous genes encoding SCM-1alpha and SCM-1beta. Here we examined the regulatory mechanism of the SCM-1 genes. The luciferase reporter gene under the control of the 5' flanking region of 0.7 kb was strongly induced upon activation with anti-CD3 or PHA plus PMA only in SCM-1-producer T cell lines through a cyclosporin A-sensitive mechanism. An element termed E1 located at -108 to -95 nt relative to the major transcription start site was found to be critical for the promoter activity. In electrophoretic mobility shift assays using the E1 oligonucleotide as probe, nuclear extracts from unstimulated T and B cell lines formed a constitutive complex termed complex I, while nuclear extracts from stimulated SCM-1-producer T cell lines formed a higher mobility complex termed complex II with a concomitant decrease in complex I. The shift from complex I to complex II seen only in SCM-1-producer T cell lines upon activation was completely suppressed by cyclosporin A. Both complexes were critically dependent on the NF-AT core sequence TTTCC in the E1 element and were partially supershifted by anti-NF-ATp. One-hybrid assays in yeast isolated NF-ATp as an E1 binding protein, and transfection of NF-ATp into T and B cell lines strongly enhanced the activation-dependent SCM-1 promoter activity. Collectively, a unique mechanism involving NF-ATp appears to regulate the cell type-specific and activation-dependent expression of the SCM-1 genes. PMID- 10477600 TI - The importance of the light chain for the epitope specificity of human anti-U1 small nuclear RNA autoantibodies present in systemic lupus erythematosus patients. AB - Abs to U1 RNA are frequently found in patients suffering from systemic lupus erythematosus overlap syndromes and Ab titers correlate with disease activity. We describe the isolation of the first human anti-U1 RNA autoantibodies from a combinatorial IgG library made from the bone marrow of a systemic lupus erythematosus patient. With the use of phage display technology, two anti-U1 RNA single-chain variable fragment (scFv) Abs were selected. Both high affinity anti U1 RNA Ab fragments (Kd approximately 1 nM) recognize stem II of U1 RNA and were derived from the same heavy chain gene (VH3-11) and the same lambda (3r) light chain gene although somatic mutations, predominantly present in the complementarity-determining regions, are different. Experiments, in which the heavy chain genes of both anti-U1 RNA scFvs were reshuffled with the original light chain repertoire of the patient resulted, after selection on stem loop II, in a large number of RNA-binding Ab fragments. All these stem loop II-specific RNA binding clones used a similar, but not identical, 3r lambda light chain. When scFvs were selected from the reshuffled libraries by stem loop IV, representing the other autoantigenic site of U1 RNA, most selected Ab clones did react with stem loop IV, but no longer with stem loop II. The stem loop IV-reactive Ab clones contained different, not 3r-related, light chains. These results point to a major role for the light chain in determining the sequence specificity of these disease-related anti-U1 RNA Abs. The possibility that secondary light chain rearrangements are involved in this autoimmune response is discussed. PMID- 10477601 TI - B lymphocyte selection and age-related changes in VH gene usage in mutant Alicia rabbits. AB - Young Alicia rabbits use VHa-negative genes, VHx and VHy, in most VDJ genes, and their serum Ig is VHa negative. However, as Alicia rabbits age, VHa2 allotype Ig is produced at high levels. We investigated which VH gene segments are used in the VDJ genes of a2 Ig-secreting hybridomas and of a2 Ig+ B cells from adult Alicia rabbits. We found that 21 of the 25 VDJ genes used the a2-encoding genes, VH4 or VH7; the other four VDJ genes used four unknown VH gene segments. Because VH4 and VH7 are rarely found in VDJ genes of normal or young Alicia rabbits, we investigated the timing of rearrangement of these genes in Alicia rabbits. During fetal development, VH4 was used in 60-80% of nonproductively rearranged VDJ genes, and VHx and VHy together were used in 10-26%. These data indicate that during B lymphopoiesis VH4 is preferentially rearranged. However, the percentage of productive VHx- and VHy-utilizing VDJ genes increased from 38% at day 21 of gestation to 89% at birth (gestation day 31), whereas the percentage of VH4 utilizing VDJ genes remained at 15%. These data suggest that during fetal development, either VH4-utilizing B-lineage cells are selectively eliminated, or B cells with VHx- and VHy-utilizing VDJ genes are selectively expanded, or both. The accumulation of peripheral VH4-utilizing a2 B cells with age indicates that these B cells might be selectively expanded in the periphery. We discuss the possible selection mechanisms that regulate VH gene segment usage in rabbit B cells during lymphopoiesis and in the periphery. PMID- 10477602 TI - The destabilization of IL-2 mRNA by a premature stop codon and its differential stabilization by trans-acting inhibitors of protein synthesis do not support a role for active translation in mRNA stability. AB - To investigate the role that translation plays in the stabilization of the IL-2 mRNA, we inhibited protein synthesis in both cis and trans. To block translation in trans, we utilized the inhibitors puromycin (PUR) and cycloheximide (CHX), which differentially effect polysome structure. We found that CHX enhances the stability of IL-2 mRNA in cells stimulated with anti-TCR Ab alone, but it inhibits CD28-induced message stabilization in costimulated cells. In contrast, PUR had a minimal effect on IL-2 mRNA stability in either the presence or absence of costimulation. The differential effects of these two inhibitors suggest that: 1) CHX is unlikely to stabilize the IL-2 mRNA by inhibiting the expression of a labile RNase; 2) CD28-mediated IL-2 mRNA stabilization does not require translation; and 3) IL-2 mRNA decay is not coupled to translation. To block translation in cis, we generated sequence-tagged IL-2 genomic reporters that contain a premature termination codon (PTC). In both the presence and absence of costimulation, these PTC-containing mRNAs exhibit drastically diminished stability. Interestingly, the addition of CHX but not PUR completely restored CD28-mediated stabilization, suggesting that CHX can block the enhanced decay induced by a PTC. Finally, CHX was able to superinduce IL-2 mRNA levels in anti TCR Ab-stimulated cells but not in CD28-costimulated cells, suggesting that CHX may also act by other mechanisms. PMID- 10477604 TI - Role of CD28 in the generation of effector and memory responses required for resistance to Toxoplasma gondii. AB - CD28 deficient (CD28-/-) mice were used to study the role of costimulation in the T cell-mediated, IFN-gamma-dependent mechanism of resistance to Toxoplasma gondii. These mice were resistant to infection with the ME49 strain of T. gondii. Analysis of the immune response of acutely infected CD28-/- mice revealed that IL 12 was required for T cell production of IFN-gamma and this was independent of the CD40/CD40 ligand interaction. A similar mechanism of IL-12-dependent, CD28/B7 independent production of IFN-gamma by T cells was also observed in wild-type mice. Interestingly, although chronically infected wild-type mice were resistant to rechallenge with the virulent RH strain of T. gondii, chronically infected CD28-/- mice were susceptible to rechallenge with the RH strain. This deficiency in the protective memory response by CD28-/- mice correlated with a lack of IL-2 and IFN-gamma in recall responses and reduced numbers of CD4+ T cells expressing a memory phenotype. Together, our findings demonstrate that CD28 is not required for the development of a protective T cell response to T. gondii, but CD28 is required for an optimal secondary immune response. PMID- 10477603 TI - A novel role for HEB downstream or parallel to the pre-TCR signaling pathway during alpha beta thymopoiesis. AB - TCR gene rearrangement and expression are central to the development of clonal T lymphocytes. The pre-TCR complex provides the first signal instructing differentiation and proliferation events during the transition from CD4-CD8-TCR- double negative (DN) stage to CD4+CD8+ double positive (DP) stage. How the pre TCR signal leads to downstream gene expression is not known. HeLa E-box binding protein (HEB), a basic helix-loop-helix transcription factor, is abundantly detected in thymocytes and is thought to regulate E-box sites present in many T cell-specific gene enhancers, including TCR-alpha, TCR-beta, and CD4. Targeted disruption of HEB results in a 5- to 10-fold reduction in thymic cellularity that can be accounted for by a developmental block at the DN to DP stage transition. Specifically, a dramatic increase in the CD4low/-CD8+CD5lowHSA+TCRlow/- immature single positive population and a concomitant decrease in the subsequent DP population are observed. Adoptive transfer test shows that this defect is cell autonomous and restricted to the alpha beta T cell lineage. Introduction of an alpha beta TCR transgene into the HEBko/ko background is not sufficient to rescue the developmental delay. In vivo CD3 cross-linking analysis of thymocytes indicates that TCR signaling pathway in the HEBko/ko mice appears intact. These findings suggest an essential function of HEB in early T cell development, downstream or parallel to the pre-TCR signaling pathway. PMID- 10477605 TI - The CD28/B7 interaction is not required for resistance to Toxoplasma gondii in the brain but contributes to the development of immunopathology. AB - Infection of C57BL/6 mice with Toxoplasma gondii leads to chronic encephalitis characterized by infiltration into the brain of T cells that produce IFN-gamma and mediate resistance to the parasite. Our studies revealed that expression of B7.1 and B7.2 was up-regulated in brains of mice with toxoplasmic encephalitis (TE). Because CD28/B7-mediated costimulation is important for T cell activation, we assessed the contribution of this interaction to the production of IFN-gamma by T cells from brains and spleens of mice with TE. Stimulation of splenocytes with Toxoplasma Ag or anti-CD3 mAb resulted in production of IFN-gamma, which was inhibited by 90% in the presence of CTLA4-Ig, an antagonist of B7 stimulation. However, production of IFN-gamma by T cells from the brains of these mice was only slightly reduced (20%) by the addition of CTLA4-Ig. To address the role of the CD28/B7 interaction during TE, we compared the development of disease in C57BL/6 wild-type (wt) and CD28-/- mice. Although the parasite burden was similar in wt and CD28-/- mice, CD28-/- mice developed less severe encephalitis and survived longer than wt mice. Ex vivo recall responses revealed that mononuclear cells isolated from the brains of chronically infected CD28-/- mice produced less IFN-gamma than wt cells, and this correlated with reduced numbers of intracerebral CD4+ T cells in CD28-/- mice compared with wt mice. Taken together, our data show that resistance to T. gondii in the brain is independent of CD28 and suggest a role for CD28 in development of immune-mediated pathology during TE. PMID- 10477606 TI - Receptor-mediated modulation of murine mast cell function by alpha-melanocyte stimulating hormone. AB - The proopiomelanocortin (POMC)-derived neuropeptide alpha-melanocyte stimulating hormone (alpha-MSH) is known to modulate some aspects of inflammation through direct effects on T cells, B cells, and monocytes. To determine whether alpha-MSH might similarly influence mast cell responsiveness, mast cells were examined to see if they expressed the receptor for alpha-MSH, melanocortin-1 (MC-1), and whether alpha-MSH altered mast cell function. We thus first identified MC-1 on bone marrow cultured murine mast cells (BMCMC) and a murine mast cell line (MCP 5) employing flow cytometry and through detection of specific binding. Subsequent treatment of mast cells with alpha-MSH increased the cAMP concentration in a characteristic biphasic pattern, demonstrating that alpha-MSH could affect intracellular processes. We next examined the effect of alpha-MSH on mediator release and cytokine expression. IgE/DNP-human serum albumin-stimulated histamine release from mast cells was inhibited by approximately 60% in the presence of alpha-MSH. Although activation of BMCMC induced the expression of mRNAs for the inflammatory cytokines IL-1beta, IL-4, IL-6, TNF-alpha, and the chemokine lymphotactin, mRNAs for IL-1beta, TNF-alpha, and lymphotactin were down-modulated in the presence of alpha-MSH. Finally, IL-3-dependent proliferative activity of BMCMC was slightly but significantly augmented by alpha-MSH. Taken together, these observations suggest that alpha-MSH may exert an inhibitory effect on the mast cell-dependent component of a specific inflammatory response. PMID- 10477607 TI - Visualization of polyoma virus-specific CD8+ T cells in vivo during infection and tumor rejection. AB - T cells are critical for clearing infection and preventing tumors induced by polyoma virus, a natural murine papovavirus. We previously identified the immunodominant epitope for polyoma virus-specific CTL in tumor-resistant H-2k mice as the Dk-restricted peptide, MT389-397, derived from the polyoma middle T oncoprotein. In this study, we developed tetrameric Dk complexes containing the MT389-397 peptide to directly visualize and enumerate MT389-397-specific CTL during polyoma virus infection. We found that Dk/MT389 tetramer+CD8+ T cells undergo a massive expansion during primary infection such that by day 7 postinfection these Ag-specific CD8+ T cells constitute approximately 20% of the total and approximately 40% of the activated CD8+ T cells in the spleen. This expansion of Dk/MT389 tetramer+CD8+ T cells parallels the emergence of MT389-397 specific ex vivo cytolytic activity and clearance of polyoma virus. Notably, Dk/MT389 tetramer+CD8+ T cells are maintained in memory at very high levels. The frequencies of Dk/MT389 tetramer+CD8+ effector and memory T cells in vivo match those of CD8+ T cells producing intracellular IFN-gamma after 6-h in vitro stimulation by MT389-397 peptide. Consistent with preferential Vbeta6 expression by MT389-397-specific CD8+CTL lines and clones, Dk/MT389 tetramer+CD8+ T cells exhibit biased expression of this Vbeta gene segment. Finally, we show that Dk/MT389 tetramer+CD8+ T cells efficiently infiltrate a polyoma tumor challenge to virus-immune mice. Taken together, these findings strongly implicate virus induced MT389-397-specific CD8+ T cells as essential effectors in eliminating polyoma-infected and polyoma-transformed cells in vivo. PMID- 10477608 TI - Inverted immunodominance and impaired cytolytic function of CD8+ T cells during viral persistence in the central nervous system. AB - Mice infected with the neurotropic JHM strain of mouse hepatitis virus (JHMV) clear infectious virus; nevertheless, virus persists in the CNS as noninfectious RNA, resulting in ongoing primary demyelination. Phenotypic and functional analysis of CNS infiltrating cells during acute infection revealed a potent regional CD8+ T cell response comprising up to 50% virus-specific T cells. The high prevalence of virus-specific T cells correlated with ex vivo cytolytic activity and efficient reduction in viral titers. Progressive viral clearance coincided with the loss of cytolytic activity, but retention of IFN-gamma secretion and increased expression of the early activation marker CD69, indicating differential regulation of effector function. Although the total number of infiltrating T cells declined following clearance of infectious virus, CD8+ T cells, both specific for the dominant viral epitopes and of unknown specificity, were retained within the CNS, suggesting an ongoing T cell response during persistent CNS infection involving a virus-independent component. Reversed immunodominance within the virus-specific CD8+ T cell population further indicated epitope-specific regulation, supporting ongoing T cell activation. Even in the absence of infectious virus, the CNS thus provides an environment that maintains both unspecific and Ag-specific CD8+ T cells with restricted effector function. Chronic T cell stimulation may thus play a role in preventing viral recrudescence, while increasing the risk of pathological conditions, such as demyelination. PMID- 10477609 TI - Expression of the protein kinase C substrate pleckstrin in macrophages: association with phagosomal membranes. AB - Despite evidence suggesting that protein kinase C (PKC) isoforms are important in phagocytosis by Fcgamma receptors, the mechanisms by which the substrates of these kinases act are largely unknown. We have investigated the role of one PKC substrate, pleckstrin, in cells of the monocyte/macrophage lineage. Pleckstrin expression in mouse macrophages was induced severalfold in response to bacterial LPS and IFN-gamma. In unstimulated cells, the protein was largely confined to the cytosol. Upon ingestion of IgG-opsonized zymosan particles (OPZ), however, pleckstrin accumulated on the phagosomal membrane. This association was transient, being maximal after 15 min and declining thereafter. Similar kinetics of association was also seen for both filamentous actin and the delta isoform of PKC. Ingestion of OPZ was found to induce phosphorylation of pleckstrin. To examine whether phosphorylation was required for phagosomal association, pleckstrin was expressed in CHO-IIA cells that stably express the FcgammaRIIA receptor and are competent for phagocytosis of OPZ. In these cells, both wild type pleckstrin and mutants in which the phosphoacceptor sites had been mutated to either alanine (nonphosphorylatable) or glutamine (pseudophosphorylated) were found to accumulate on OPZ phagosomes. Thus, association of pleckstrin with phagosomes is independent of its phosphorylation. Our findings suggest that pleckstrin may serve as an intracellular adaptor/targeting protein in response to particulate stimuli. By targeting interacting ligands to the phagosomal compartment, pleckstrin may serve to regulate phagocytosis and/or early steps during maturation of the phagosome. PMID- 10477610 TI - Fc-mediated nonspecific binding between fibronectin-binding protein I of Streptococcus pyogenes and human immunoglobulins. AB - Fibronectin-binding protein I (SfbI) from Streptococcus pyogenes plays a key role in bacterial adhesion to, and invasion of, eukaryotic cells. In addition, SfbI exhibits a considerable potential as mucosal adjuvant and can trigger polyclonal activation of B cells. Here, we report that SfbI is also capable of binding human IgG in a nonimmune fashion. SfbI was reactive with IgG1, IgG2, IgG3, and IgG4 isotypes (type IIo IgG-binding profile). The affinity constant (Kd) of the SfbI IgG interaction was in the range of 1-2 x 10(-5) M. Further studies demonstrated that the SfbI binding was mediated by the Fc component of the IgG molecule. Experiments performed using purified recombinant proteins spanning different domains of SfbI showed that the IgG-binding activity was restricted to the fibronectin-binding domains, and in particular to the fibronectin-binding repeats. Finally, the presence of recombinant SfbI resulted in an impairment of both phagocytosis of IgG-coated RBCs and Ab-dependent cell cytotoxicity by macrophages. These results demonstrated for the first time that, in addition to its major role during the colonization process, SfbI may also favor bacterial immune evasion after the onset of the infection by interfering with host clearance mechanisms. PMID- 10477611 TI - Requirement for CD4+ T cells in V beta 4+CD8+ T cell activation associated with latent murine gammaherpesvirus infection. AB - A CD8+ T cell lymphocytosis in the peripheral blood is associated with the establishment of latency following intranasal infection with murine gammaherpesvirus-68. Remarkably, a large percentage of the activated CD8+ T cells of mice expressing different MHC haplotypes express V beta 4+ TCR. Identification of the ligand driving the V beta 4+CD8+ T cell activation remains elusive, but there is a general correlation between V beta 4+CD8+ T cell stimulatory activity and establishment of latency in the spleen. In the current study, the role of CD4+ T cells in the V beta 4+CD8+ T cell expansion has been addressed. The results show that CD4+ T cells are essential for expansion of the V beta 4+CD8+ subset, but not other V beta subsets, in the peripheral blood. CD4+ T cells are required relatively late in the antiviral response, between 7 and 11 days after infection, and mediate their effect independently of IFN-gamma. Assessment of V beta 4+CD8+ T cell stimulatory activity using murine gammaherpesvirus-68-specific T cell hybridomas generated from latently infected mice supports the idea that CD4+ T cells control levels of the stimulatory ligand that drives the V beta 4+CD8+ T cells. As V beta 4+CD8+ T cell expansion also correlates with levels of activated B cells, these data raise the possibility that CD4+ T cell-mediated B cell activation is required for optimal expression of the stimulatory ligand. In addition, in cases of low ligand expression, there may also be a direct role for CD4+ T cell-mediated help for V beta 4+CD8+ T cells. PMID- 10477612 TI - A monoclonal antibody directed against the murine macrophage surface molecule F4/80 modulates natural immune response to Listeria monocytogenes. AB - Whole spleen cell cultures from SCID mice release high levels of IFN-gamma when exposed to heat-killed Listeria monocytogenes (HKL). This microbe-induced and T cell-independent response depends on both macrophages (MPhi) and NK cells: HKL stimulated MPhi release TNF-alpha and IL-12, which together activate NK cells for IFN-gamma release. We show here that this cytokine-mediated activation cascade can be modulated by a mAb against the MPhi surface glycoprotein F4/80. HKL induced IL-12, TNF-alpha, and IFN-gamma in SCID whole spleen cell cultures was inhibited by coincubation with anti-F4/80 mAb whereas IL-1 and IL-10 were enhanced. Both effects were apparent at mRNA and protein release levels. Whereas inhibitory activities were F4/80 Ag specific, stimulatory effects were Fc dependent and nonspecific. Furthermore, cytokine inhibition by anti-F4/80 was only apparent when MPhi and NK cells were present simultaneously and in close vicinity, indicating that direct cell-to-cell contact is a prerequisite. These data suggest a novel pathway for microbe-induced MPhi/NK cell interaction involving direct cell-to-cell signaling and give the first evidence for a functional role of the MPhi surface glycoprotein F4/80. PMID- 10477613 TI - Oral administration of lipopolysaccharide exacerbates collagen-induced arthritis in mice. AB - We investigated whether oral administration of LPS exacerbated collagen-induced arthritis (CIA) in mice, which was an experimental model of autoimmune disease. CIA was induced by s.c. injection of type II collagen emulsified with CFA into the base of the tail (day 0) followed by a booster injection on day 21. To examine the ability of LPS to exacerbate CIA, varying doses of LPS were orally administered on day 50. The results showed that administration of LPS was followed by reactivation of CIA in a dose-related fashion. Histologically, on day 55 there were marked edema of synovium proliferated by day 50, new formation of fibrin, and intense infiltration of neutrophils accompanied with a large number of mononuclear cells. Severe destruction of cartilage and subchondral bone was also observed on day 70. The reactivation of CIA by oral administration of LPS was associated with increase in anti-type II collagen IgG and IgG2a Abs as well as varying kinds of cytokines including IL-12, IFN-gamma, IL-1beta, and TNF alpha. Polymyxin B sulfate given either orally or i.v. suppressed the recurrence of CIA. Increased amounts of LPS were found in sera of mice given the endotoxin orally. LPS from Salmonella enteritidis, Salmonella typhimurium, and Klebsiella pneumoniae and its component, lipid A from Escherichia coli, also reactivated the disease. These findings suggest that LPS from intestinal bacteria may play a role in the exacerbation of autoimmune joint inflammation. PMID- 10477614 TI - Cytosolic phospholipase A2 activation is essential for beta 1 and beta 2 integrin dependent adhesion of human eosinophils. AB - We examined the role of cytosolic phospholipase A2 (cPLA2) during human eosinophil adherence to ICAM-1- or VCAM-1-coated plates. IL-5-stimulated eosinophils adhered to ICAM-1 through the beta 2 integrin CD11b/CD18, while nonstimulated eosinophils did not. By contrast, nonstimulated eosinophils adhered to VCAM-1 through the beta 1-integrin VLA-4/CD29. Both IL-5-induced adhesion to ICAM-1 and spontaneous adhesion to VCAM-1 corresponded temporally to cPLA2 phosphorylation, which accompanied enhanced catalytic activity of cPLA2. The structurally unrelated cPLA2 inhibitors, arachidonyl trifluoromethylketone and surfactin, significantly inhibited eosinophil adhesion to ICAM-1 and VCAM-1 in a concentration-dependent manner. Inhibition of secretory PLA2, 5-lipoxygenase, or cyclooxygenase did not affect eosinophil adhesion. Addition of arachidonic acid to eosinophils after cPLA2 inhibition with arachidonyl trifluoromethylketone or surfactin did not reverse the blockade of adhesion to ICAM-1 or VCAM-1. However, CV-6209, a receptor-specific antagonist of platelet-activating factor, inhibited all integrin-mediated adhesion. The activated conformation of CD11b as identified by the mAb, CBRM1/5, as well as quantitative surface CD11b expression were up regulated after IL-5 stimulation. However, cPLA2 inhibition neither prevented CBRM1/5 expression nor blocked surface Mac-1 up-regulation caused by IL-5. Our data suggest that cPLA2 activation and its catalytic product platelet-activating factor play an essential role in regulating beta 1 and beta 2 integrin-dependent adhesion of eosinophils. This blockade occurs even in the presence of up regulated eosinophil surface integrin. PMID- 10477615 TI - The nonintegrin laminin binding protein (p67 LBP) is expressed on a subset of activated human T lymphocytes and, together with the integrin very late activation antigen-6, mediates avid cellular adherence to laminin. AB - A search for genes expressed in activated T cells revealed that the nonintegrin, 67-kDa laminin binding protein (p67 LBP) is expressed on the surface of a subset (10-15%) of activated peripheral blood T cells. Surface p67 LBP expression is detectable by FACS using the anti-p67 LBP mAb, MLuC5, within 6 h of T cell activation with phorbol dibutyrate and ionomycin, peaks 18-36 h postactivation, and persists for 7-10 days. The subset of T cells expressing p67 LBP is composed of mature, single-positive cells (85% CD4+8-, 15% CD4-8+) of memory cell phenotype (100% CD45 RO+/CD45 RA-). The p67 LBP+ T cells also express the integrin alpha6 chain (CD49f), which is known to associate with p67 LBP on tumor cells. In addition, the p67 LBP+ T cells express the integrin beta1, which associates with alpha6 in the laminin-specific integrin receptor very late activation Ag (VLA)-6 (alpha6beta1). Expression of an exogenous cDNA encoding the 37-kDa LBP precursor (p37 LBPP) confers p67 LBP surface expression on a p67 LBP negative Jurkat T cell line (B2.7). Expression of p67 LBP induces B2.7 transfectants to adhere to laminin, but avid laminin binding depends on coexpression of VLA-6. Taken together, these data indicate that p67 LBP is an activation-induced surface structure on memory T cells that, together with VLA-6, mediates cellular adherence to laminin. PMID- 10477616 TI - Molecular mechanisms underlying IL-4-induced leukocyte recruitment in vivo: a critical role for the alpha 4 integrin. AB - IL-4 is known to induce recruitment of eosinophils and mononuclear leukocytes. In vitro this occurs in part by selective expression of VCAM-1, the ligand for the alpha 4 integrin. The objective of this study was to determine the molecular mechanisms that underlie IL-4-induced leukocyte recruitment in vivo. Mice received an intrascrotal injection of IL-4 (100 ng). Twenty-four hours later, leukocyte rolling, adhesion, and emigration in cremasteric postcapillary venules were examined via intravital microscopy, and expression of VCAM-1 and P- and E selectin was quantitated using a radiolabeled mAb technique. IL-4 increased VCAM 1 expression, but P-selectin and E-selectin remained at constitutive levels. IL-4 induced significant increases in leukocyte adhesion and emigration, with 50% of the emigrated cells being eosinophils and the remainder being mononuclear leukocytes. Leukocyte rolling in IL-4-treated mice was >95% inhibitable using an anti-P-selectin Ab. However, IL-4-induced leukocyte recruitment was unaltered in mice treated chronically with P-selectin Ab or mice deficient in either P selectin or P- and E-selectin, suggesting that the residual rolling supported all of the IL-4-induced recruitment. In IL-4-treated mice following P-selectin blockade, tethering and rolling were not dependent on L-selectin, but were abolished by alpha 4 integrin blockade. These findings show that the alpha 4 integrin can initiate leukocyte-endothelial cell interactions in the absence of selectins under shear conditions in vivo, and that the absence of selectins does not affect recruitment of eosinophils and mononuclear cells to IL-4-treated tissue. PMID- 10477617 TI - Role of 5-lipoxygenase products in the local accumulation of neutrophils in dermal inflammation in the rabbit. AB - Studies were undertaken to define the role of 5-lipoxygenase (5-LO) products and, in particular, of leukotriene (LT) B4 in the polymorphonuclear leukocyte (PMN) emigration process using a rabbit model of dermal inflammation. Our results show that i.v. administration to rabbits of MK-0591, a compound that inhibits LT biosynthesis in blood and tissues when administered in vivo, significantly reduced 51Cr-labeled PMN accumulation in response to intradermally injected chemotactic agonists, including IL-8, FMLP, C5a, and LTB4 itself. In addition, pretreatment of the labeled PMN with MK-0591 ex vivo before their injection in recipient animals was equally effective in reducing 51Cr-labeled PMN emigration to dermal inflammatory sites. These results support a role for de novo synthesis of 5-LO metabolites by PMN for their chemotactic response to inflammatory mediators. Other studies demonstrated that elevated intravascular concentration of LTB4 interferes with PMN extravasation inasmuch as a continuous i.v. infusion of LTB4, in the range of 5-300 ng/min/kg, dose-dependently inhibited extravascular PMN accumulation to acute inflammatory skin sites elicited by the chemoattractants LTB4, FMLP, C5a, and IL-8 and by TNF-alpha, IL-1beta, and LPS; such phenomena may constitute a natural protective mechanism from massive tissue invasion by activated PMN in specific pathologic conditions such as ischemia (and reperfusion). These studies demonstrate additional functions of 5-LO products in the regulation of PMN trafficking, distinct from the well-characterized chemotactic activity of LTB4 present in the extravascular compartment. PMID- 10477618 TI - A novel mechanism of action of chemically modified tetracyclines: inhibition of COX-2-mediated prostaglandin E2 production. AB - Tetracyclines (doxycycline and minocycline) inhibit inducible NO synthase expression and augment cyclooxygenase (COX)-2 expression and PGE2 production. In contrast, chemically modified tetracyclines (CMTs), such as CMT-3 and -8 (but not CMT-1, -2, and -5), that lack antimicrobial activity, inhibit both NO and PGE2 production in LPS-stimulated murine macrophages, bovine chondrocytes, and human osteoarthritis-affected cartilage, which spontaneously produces NO and PGE2 in ex vivo conditions. Furthermore, CMT-3 augments COX-2 protein expression but inhibits net PGE2 accumulation. This coincides with the ability of CMT-3 and -8 to inhibit COX-2 enzyme activity in vitro. The action of CMTs is distinct from that observed with tetracyclines because 1) CMT-3-mediated inhibition of PGE2 production coincides with modification of COX-2 protein, which is distinct from the nonglycosylated COX-2 protein generated in the presence of tunicamycin, as observed by Western blot analysis and 2) CMT-3 and -8 have no significant effect on COX-2 mRNA accumulation. In contrast, CMT-3 and -8 do not inhibit COX-1 expression in A549 human epithelial cells at the level of protein and mRNA accumulation or modification of COX-1 protein. CMT-3 and -8 inhibit the sp. act. of COX-2 (but not COX-1) in cell-free extracts. These results demonstrate differential action of CMT-3 (Metastat) on COX-1 and -2 expression, which is distinct from other tetracyclines. PMID- 10477619 TI - A defective protein kinase C anchoring system underlying age-associated impairment in TNF-alpha production in rat macrophages. AB - The ability of macrophages to secrete cytokines is important in host responses to infections inflammatory stimuli, both of which are altered with aging. In this study, age-associated changes in the release of TNF-alpha from LPS-stimulated rat alveolar macrophages were determined and correlated with a decrease in the level of RACK1, the anchoring protein involved in protein kinase C translocation and activation. Macrophages from aged rats produced approximately 50% less TNF-alpha than those from young rats. This effect was observed independently from the concentration of LPS used and the time considered. The decrease observed was associated with a defective PKC translocation, due to a reduction in the expression of RACK1, whereas no differences were detected in the expression of LPS receptor (CD14) or total PKC isoforms (alpha and betaIotaIota) in old and young rats. Use of RACK1 antisense oligonucleotide reduced the ability of young macrophages to respond to LPS, further supporting the idea that a deficit in RACK1 contributes to the functional impairment in aged macrophages and that age induced macrophage immunodeficiencies are associated with alteration in signal transduction pathways. PMID- 10477620 TI - Curcumin blocks cytokine-mediated NF-kappa B activation and proinflammatory gene expression by inhibiting inhibitory factor I-kappa B kinase activity. AB - NF-kappa B plays a critical role in the transcriptional regulation of proinflammatory gene expression in various cells. Cytokine-mediated activation of NF-kappa B requires activation of various kinases, which ultimately leads to the phosphorylation and degradation of I kappa B, the NF-kappa B cytoplasmic inhibitor. The food derivative curcumin has been shown to inhibit NF-kappa B activity in some cell types. In this report we investigate the mechanism of action of curcumin on cytokine-induced proinflammatory gene expression using intestinal epithelial cells (IEC). Curcumin inhibited IL-1 beta-mediated ICAM-1 and IL-8 gene expression in IEC-6, HT-29, and Caco-2 cells. Cytokine-induced NF kappa B DNA binding activity, RelA nuclear translocation, I kappa B alpha degradation, I kappa B serine 32 phosphorylation, and I kappa B kinase (IKK) activity were blocked by curcumin treatment. Wound-induced p38 phosphorylation was not inhibited by curcumin treatment. In addition, mitogen-activated protein kinase/ERK kinase kinase-1-induced IL-8 gene expression and 12-O-tetraphorbol 12 myristate 13-acetate-responsive element-driven luciferase expression were inhibited by curcumin. However, I kappa B alpha degradation induced by ectopically expressed NF-kappa B-inducing kinase or IKK was not inhibited by curcumin treatment. Therefore, curcumin blocks a signal upstream of NF-kappa B inducing kinase and IKK. We conclude that curcumin potently inhibits cytokine mediated NF-kappa B activation by blocking a signal leading to IKK activity. PMID- 10477621 TI - Glucocorticoids induce apoptosis in human monocytes: potential role of IL-1 beta. AB - Glucocorticoids (GC) are potent anti-inflammatory and immunosuppressive agents that act on a variety of immune cells, including monocytes and macrophages. However, the exact cellular mechanisms underlying this anti-inflammatory capacity are still unknown. In our study, we determined the induction of apoptosis by GC in human monocytes. Peripheral blood monocytes were isolated by density centrifugation methods with a purity of >90% and were cultured in RPMI 1640 medium. Monocyte apoptosis was determined by four independent methods, including annexin-V staining, TUNEL, DNA-laddering, and typical morphology by means of transmission electron microscopy. TNF-alpha and IL-1beta were measured by ELISA. GC receptor was blocked with mifepristone. Caspase 3 was inhibited with caspase-3 inhibitor (DEVD-CHO). Stimulation with different GC at therapeutic concentrations resulted in monocyte apoptosis in a time- and dose-dependent manner. Necrosis was excluded by propidium iodide staining. Proinflammatory cytokines such as IL-1beta and TNF-alpha were down-regulated by GC treatment. Continuous treatment of monocytes with IL-1beta, but not with TNF-alpha, could almost completely prevent GC-induced cell death. The addition of mifepristone or caspase-3 inhibitor could partially abrogate GC-induced apoptosis as well as GC-induced inhibition of IL 1beta. This is the first study to demonstrate induction of apoptosis by GC in human monocytes. GC-induced monocyte apoptosis may be partially mediated through effects on IL-1beta production. It is conceivable that GC exert their anti inflammatory capacity in various diseases, at least in part, by the induction of apoptosis in monocytes. PMID- 10477622 TI - A deficiency of placental IL-10 in preeclampsia. AB - Accommodation of the fetoplacental unit in human pregnancy requires maternal immune tolerance to this "semiallograft". Local antiplacental immunity is modified by synthesis of uncommon histocompatibility Ags (e.g., HLA-G), growth factors, and cytokines by the placenta. Placental interleukins have been identified in reproductive tissues, but their roles in adaptive maternal immunity and determining term pregnancy outcomes have not been fully clarified. This study examined the distribution of IL-10 and TNF-alpha staining in term placentas. Women with proteinuric hypertension (PE, n = 10) were compared with an age matched group with normal pregnancy (NP, n = 14) and gestational hypertension (GH, n = 6). Using immunohistochemistry of parrafin-fixed tissues, trophoblast cells were identified by cytokeratin 7 and cytokeratin 18 staining. The cytokine binding of villous trophoblast cells was scored depending on the extent of circumferential cytoplasm staining (<25%; intermediate or >75%). The cytokine positive decidual cells were scored as a percentage of total extravillous trophoblast cells. There was a reduction in villous IL-10 immunostaining compared with normal term placenta (PE, 10.2 +/- 1.1, mean +/- SEM; NP, 14.07 +/- 1.16 Mann-Whitney U test; p = 0.02). In these patients, there was an increase in TNF alpha immunostaining. Sparse endovascular extravillous trophoblast cells demonstrated nuclear IL-10 staining in 30% of patients with preeclampsia. Serum IL-10 was diminished in women with preeclampsia compared with normal pregnancy. In conclusion, villous trophoblast demonstrated diminished immunostaining of IL 10 in preeclampsia. This abnormality may be associated with heightened maternal antifetal immunity and therefore inadequate placental development in preeclampsia. PMID- 10477623 TI - Cloning the antibody response in humans with inflammatory central nervous system disease: analysis of the expressed IgG repertoire in subacute sclerosing panencephalitis brain reveals disease-relevant antibodies that recognize specific measles virus antigens. AB - The presence of increased IgG in the brains of humans with infectious and inflammatory CNS diseases of unknown etiology such as multiple sclerosis may be a clue to the cause of disease. For example, the intrathecally synthesized oligoclonal bands (OGBs) in diseases such as subacute sclerosing panencephalitis (SSPE) or cryptococcal meningitis have been shown to represent Ab directed against the causative agents, measles virus (MV) or Cryptococcus neoformans, respectively. Using SSPE as a model system, we have developed a PCR-based strategy to analyze the repertoire of IgG V region sequences expressed in SSPE brain. We observed abnormal expression of germline V segments, overrepresentation of particular sequences that correspond to the oligoclonal bands, and substantial somatic mutation of most clones from the germline, which, taken together, constitute features of Ag-driven selection in the IgG response. Using the most abundant or most highly mutated gamma H chain and kappa or lambda L chain sequences in various combinations, we constructed functional Abs in IgG mammalian expression vectors. Three Abs specifically stained MV-infected cells. One Ab also stained cells transfected with the MV nucleoprotein, and a second Ab stained cells transfected with the MV-fusion protein. This technique demonstrates that functional Abs produced from putative disease-relevant IgG sequences can be used to recognize their corresponding Ags. PMID- 10477624 TI - Enhanced autoimmune arthritis in IFN-gamma receptor-deficient mice is conditioned by mycobacteria in Freund's adjuvant and by increased expansion of Mac-1+ myeloid cells. AB - Induction of experimental autoimmune diseases often relies on immunization with the organ-specific autoantigens in CFA, which contains heat-killed mycobacteria. In several of these models, including collagen-induced arthritis, endogenous IFN gamma acts as a disease-limiting factor in the pathogenesis of the disease. Here we show that in collagen-induced arthritis the protective effect of IFN-gamma depends on the presence of mycobacteria in the adjuvant. Omission of mycobacteria inverts the role of endogenous IFN-gamma to a disease-promoting factor. Thus, the mycobacterial component of CFA opens a pathway by which endogenous IFN-gamma exerts a protective effect that supersedes its otherwise disease-promoting effect. Extramedullary hemopoiesis and expansion of the Mac-1+ cell population accompanied the accelerated and more severe disease course in the IFN-gamma receptor knockout mice immunized with CFA. Treatment of such mice with Abs against the myelopoietic cytokines IL-6 or IL-12 inhibited both disease development and the expansion of the Mac-1+ population. We postulate that mycobacteria in CFA stimulate the expansion of the Mac-1+ cell population by a hemopoietic process that is restrained by endogenous IFN-gamma. These results have important implications for the validity of animal models of autoimmunity to study the pathogenesis and to evaluate cytokine-based therapy of autoimmune diseases. PMID- 10477625 TI - Evidence that Langerhans cells in adult pulmonary Langerhans cell histiocytosis are mature dendritic cells: importance of the cytokine microenvironment. AB - Because Langerhans cells (LC) in peripheral tissues are generally "immature" cells with poor lymphostimulatory activity, the contribution of immune responses initiated by LC to the pathogenesis of pulmonary LC histiocytosis (LCH) has been uncertain. In this study we demonstrate that LC accumulating in LCH granulomas are phenotypically similar to mature lymphostimulatory dendritic cells present in lymphoid organs. LC in LCH granulomas intensely expressed B7-1 and B7-2 molecules, whereas normal pulmonary LC and LC accumulating in other pathologic lung disorders did not express these costimulatory molecules. The presence of B7+ LC in LCH granulomas was associated with the expression in these lesions, but not at other sites in the lung, of a unique profile of cytokines (presence of GM-CSF, TNF-alpha, and IL-1beta and the absence of IL-10) that is known to promote the in vitro differentiation of LC into cells expressing a lymphostimulatory phenotype. Finally, LCH granulomas were the only site where CD154-positive T cells could be identified in close contact with LC intensely expressing CD40 Ags. Taken together, these results strongly support the idea that an abnormal immune response initiated by LC may participate in the pathogenesis of pulmonary LCH, and suggest that therapeutic strategies aimed at modifying the lymphostimulatory phenotype of LC may be useful in the treatment of this disorder. PMID- 10477626 TI - Polarized type 1 cytokine profile in bronchoalveolar lavage T cells of patients with hypersensitivity pneumonitis. AB - Hypersensitivity pneumonitis (HP) is characterized by an inflammatory lymphocytic alveolitis comprised of both CD8+ and CD4+ T cells. Animal models suggest that HP is facilitated by overproduction of IFN-gamma, and that IL-10 ameliorates severity of the disease, indicating a Th1-type response. To determine whether a Th1 phenotype in HP also exists clinically, bronchoalveolar lavage (BAL) and peripheral blood (PB) T cells were obtained from HP individuals and analyzed for Th1 vs Th2 cytokine profiles. It was determined that soluble OKT3-stimulated BAL T cells cocultured with alveolar macrophages produced more IFN-gamma and less IL 10 than PB T cells cocultured with monocytes, but no difference was observed in IL-4 production. The monocytic cells did not account for this difference, as CD80 and CD86 expressions were similar, and coculturing PB T cells with alveolar macrophages resulted in no difference in IFN-gamma production. Similarly, there was no difference in IL-12 production between stimulated BAL or PB T cells; however, addition of rIL-12 significantly increased production of IFN-gamma by BAL T cells, but not by PB T cells. This effect was due to a difference in IL-12R expression. High affinity IL-12R were only present in association with BAL T cells. These studies indicate that clinical HP is characterized by a predominance of IFN-gamma-producing T cells, perhaps resulting from a reduction in IL-10 production and an increase in high affinity IL-12R compared with blood T cells. PMID- 10477627 TI - Up-regulation of CCR2 chemokine receptor expression and increased susceptibility to the multitropic HIV strain 89.6 in monocytes exposed to glucocorticoid hormones. AB - Glucocorticoid hormones (GC) are potent antiinflammatory agents widely used in the treatment of diverse human diseases. The present study was aimed at assessing the effect of GC on chemokine receptor expression in human monocytes. Dexamethasone (Dex) up-regulated mRNA expression of the monocyte chemotactic protein (MCP-1, CCL2) chemokine receptor CCR2. The effect was selective in that other chemokine receptors were not substantially affected. Stimulation by Dex was observed after 4 h of exposure at concentrations of 10(-7) to 10(-5) M. Steroids devoid of GC activity were inactive, and the GC receptor antagonist, RU486, inhibited stimulation. Dex did not affect the rate of nuclear transcription, but augmented the CCR2 mRNA half-life. Augmentation of CCR2 expression by Dex was associated with increased chemotaxis. Finally, Dex treatment induced productive replication of the HIV strain 89.6, which utilizes CCR2 as entry coreceptor, in freshly isolated monocytes. Together with previous findings, these results indicate that at least certain pro- and antiinflammatory molecules have reciprocal and divergent effects on expression of a major monocyte chemoattractant, MCP-1, and of its receptor (CCR2). Augmentation of monocyte CCR2 expression may underlie unexplained in vivo effects of GC as well as some of their actions on HIV infection. PMID- 10477628 TI - A common TCR V-D-J sequence in V beta 13.1 T cells recognizing an immunodominant peptide of myelin basic protein in multiple sclerosis. AB - T cell responses to the immunodominant peptide (residues 83-99) of myelin basic protein are potentially associated with multiple sclerosis (MS). This study was undertaken to examine whether a common sequence motif(s) exists within the TCR complementarity-determining region (CDR)-3 of T cells recognizing the MBP83-99 peptide. Twenty MBP83-99-reactive T cell clones derived from patients with MS were analyzed for CDR3 sequences, which revealed several shared motifs. Some V beta 13.1 T cell clones derived from different patients with MS were found to contain an identical CDR3 motif, V beta 13.1-LGRAGLTY. Oligonucleotides complementary to the shared CDR3 motifs were used as specific probes to detect identical target CDR3 sequences in a large panel of T cell lines reactive to MBP83-99 and unprimed PBMC. The results revealed that, in contrast to other CDR3 motifs examined, the LGRAGLTY motif was common to T cells recognizing the MBP83 99 peptide, as evident by its expression in the majority of MBP83-99-reactive T cell lines (36/44) and PBMC specimens (15/48) obtained from randomly selected MS patients. The motif was also detected in lower expression in some PBMC specimens from healthy individuals, suggesting the presence of low precursor frequency of T cells expressing this motif in healthy individuals. This study provides new evidence indicating that the identified LGRAGLTY motif is preferentially expressed in MBP83-99-reactive T cells. The findings have important implications in monitoring and targeting MBP83-99-reactive T cells in MS. PMID- 10477629 TI - Markers of lymphocyte homing distinguish CD4 T cell subsets that turn over in response to HIV-1 infection in humans. AB - In HIV-1 infection, the abrupt rise in CD4 T cells after effective antiretroviral therapy has been viewed as a measure of HIV-1-related CD4 T cell turnover in the steady state. The early (2-4 wk) response is reportedly dominated by CD4 T cells with a memory (CD45RO) phenotype. It is controversial whether the measurement of steady-state kinetics identifies cells that otherwise would have been recruited into a short-lived, virus-producing pool or reflects lymphoid redistribution/sequestration. We performed detailed phenotypic and kinetic analysis of CD4 T cell subsets in 14 patients. Turnover occurs in memory (CD45RO) as well as naive (CD45RA) cells, if the latter are present at baseline. Most of the turnover occurs in those memory (CD45RO) and naive (CD45RA) cells that are programmed for recirculation through lymphoid organs (CD62L+ and CD44low), whereas very little turnover occurs in memory cells (CD45RO) destined for recirculation from blood to tissue (CD62L- and CD44high). Turnover occurs in both activated (CD25+ and HLA-DR+) and nonactivated populations, although it is restricted to CD38-positive cells, indicating that turnover does not measure cells that are already infected. More likely, turnover occurs in cells that replace infected cells or are on their way to becoming infected. Taken together, markers of lymphocyte trafficking better describe cell turnover related to virus replication than do naive and memory markers per se, and lymph organs, not tissue destined cells or peripheral blood cells, appear to be the important site of virus replication and CD4 T cell turnover, destruction, and redistribution. PMID- 10477630 TI - IgG and complement-mediated tissue damage in the absence of C2: evidence of a functionally active C2-bypass pathway in a guinea pig model. AB - In vitro complement-mediated lysis of heavily sensitized sheep erythrocytes by C4 deficient (C4D) guinea pig and C2-deficient (C2D) human sera was demonstrated some years ago. It was postulated that these "complement-bypass" pathways resulted from activation of C1 and components of the alternative pathway. We used normal, C2D, and C4D guinea pigs in a Forssman shock model to test the in vivo relevance of the C2- and C4-bypass pathways of complement activation. High concentrations of both anti-Forssman Ab and C2D or C4D guinea pig serum induced efficient lysis of sheep erythrocytes in vitro. The most efficient lysis was observed when IgG Ab and C2D guinea pig serum were used. Blocking either the classical pathway (treatments with EGTA-Mg2+ or soluble recombinant complement receptor type 1 (sCR1)) or the alternative pathway (treatment with heating at 50 degrees C, sCR1, or soluble recombinant CR1 lacking the first of the four long homologous repeat sequences (sCR1[desLHR-A])) inhibited lysis; both pathways were required for lysis of sheep erythrocytes by C2D and C4D guinea pig sera. i.v. injection of anti-Forssman Ab in normal guinea pigs resulted in rapid death from pulmonary shock, whereas C4D guinea pigs had no adverse effect. Surprisingly, C2D guinea pigs either died in a delayed fashion or had a sublethal reaction. sCR1 treatment prevented Forssman shock in both normal and C2D guinea pigs, whereas sCR1[desLHR-A] prevented Forssman shock only in C2D animals. Our results suggest that the C2-bypass pathway occurs in vivo to produce tissue damage. Activation of complement in the absence of C2 appears to be far more efficient than in the absence of C4. PMID- 10477631 TI - Spinal cord ischemia after abdominal aortic operation: is it preventable? AB - PURPOSE: Spinal cord ischemia after operation on the abdominal aorta is a rare event that is attributed to variations in the spinal cord blood supply. The purpose of this study was to evaluate the possible causes of this devastating event. METHODS: A survey of patients among the members of the Southern Association for Vascular Surgery was performed, and 18 patients were identified with spinal cord ischemia manifested by paraplegia or paraparesis after abdominal aortic operation. RESULTS: Preoperative computed tomographic, magnetic resonance, and aortographic results did not visualize the greater radicular artery (Adamkiewicz's artery) in any patient. Eleven patients underwent resection of infrarenal abdominal aortic aneurysms (AAAs): seven of these patients had tube grafts, three had aortobifemoral grafts, and one had an aortobiiliac graft. Five other patients underwent placement of aortobifemoral grafts, and one patient underwent aortobiiliac graft placement for occlusive disease. One patient underwent suprarenal AAA resection with an interposition graft to a previous aortobiiliac graft. The mean operative time was 3 hours and 39 minutes (range, 2 hours and 45 minutes to 6 hours and 30 minutes), with a mean aortic cross-clamp time of 48 minutes (range, 24 to 97 minutes). Sixteen aortic cross-clamps were placed infrarenally and two suprarenally (one in a case of ruptured AAA, the other a suprarenal AAA). Seventeen proximal anastomoses were end to end. The average minimum systolic blood pressure during the aortic cross-clamping was 96 mm Hg (range, 80 to 130 mm Hg). All the patients had internal iliac artery flow preserved with either prograde perfusion (10 patients) or retrograde perfusion (eight patients), and one patient underwent unilateral internal iliac artery ligation because of aneurysmal disease. One aortobifemoral-graft limb necessitated thrombectomy, but no cases of massive peripheral embolization occurred. When paraplegia was suspected after operation (6 to 20 hours after surgery), five patients underwent lumbar drainage. No clinical improvement was noted. CONCLUSION: Interference with pelvic blood supply from prolonged aortic cross clamping, intraoperative hypotension, aortic embolization, and interruption of internal iliac artery circulation have all been suggested as possible causes of spinal cord ischemia. In this survey, none of these factors proved to be significant as the sole cause of spinal cord ischemia. In the performance of an aortic operation with an end-to-end proximal anastomosis in the presence of severe external or internal iliac artery disease, there may be an increased incidence of spinal cord ischemia despite appropriate surgical techniques to ensure internal iliac perfusion. Spinal cord ischemia after abdominal aortic operations appears to be a tragically unpredictable, random, and unpreventable event. PMID- 10477632 TI - Comparison between the transabdominal and retroperitoneal approaches for aortic reconstruction in patients at high risk. AB - PURPOSE: The purpose of this study was to compare the transabdominal approach with the retroperitoneal approach for elective aortic reconstruction in the patient who is at high risk. METHODS: From January 1992 through January 1997, 148 patients underwent aortic operations: 92 of the patients were classified as American Society of Anesthesia (ASA) class IV. Forty-four operations on the patients of ASA class IV were performed with the transabdominal approach (25 for abdominal aortic aneurysms and 19 for aortoiliac occlusive disease), and 48 operations were performed with the retroperitoneal approach (27 for abdominal aortic aneurysms and 21 for aortoiliac occlusive disease). There were no significant differences between the groups for comorbid risk factors or perioperative care. RESULTS: Among the patients of ASA class IV, eight (8.7%) died after operation (retroperitoneal, 3 [6.26%]; transabdominal, 5 [11.3%]; P =.5). There was no difference between groups in the number of pulmonary complications (retroperitoneal, 23 [47.9%]; transabdominal, 19 [43.2%]; P =.7) or in the development of incisional hernias (retroperitoneal, 6 [12.5%]; transabdominal, 5 [11.3%]; P =.5). The retroperitoneal approach was associated with a significant reduction in cardiac complications (retroperitoneal, 6 [12.5%]; transabdominal, 10 [22.7%]; P =.004) and in gastrointestinal complications (retroperitoneal, 5 [8.3%]; transabdominal, 15 [34.1%]). Operative time was significantly longer in the retroperitoneal group (retroperitoneal, 3.35 hours; transabdominal, 2.98 hours; P =.006), as was blood loss (retroperitoneal, 803 mL; transabdominal, 647 mL; P =.012). The patients in the retroperitoneal group required less intravenous narcotics (retroperitoneal, 36.6 +/- 21 mg; transabdominal, 49.5 +/- 28.5 mg; P =.004) and less epidural analgesics (retroperitoneal, 39.5 +/- 6.4 mg; transabdominal, 56.6 +/- 9.5 mg; P =.004). Hospital length of stay (retroperitoneal, 7.2 +/- 1.6 days; transabdominal, 12.8 +/- 2.3 days; P =.024) and hospital charges (retroperitoneal, $35,587 +/- $980; transabdominal, $54,832 +/- $1105; P =.04) were significantly lower in the retroperitoneal group. The survival rates at the 40-month follow-up period were similar between the groups (retroperitoneal, 81.3%; transabdominal, 78.7%; P =.53). CONCLUSION: In this subset of patients who were at high risk for aortic reconstruction, the postoperative complications were common. However, the number of complications was significantly lower in the retroperitoneal group. Aortic reconstruction in patients of ASA class IV appears to be more safely and economically performed with the retroperitoneal approach. PMID- 10477633 TI - Endovascular repair of abdominal aortic aneurysms in patients with congenital renal vascular anomalies. AB - PURPOSE: The endovascular repair of abdominal aortic aneurysms (AAAs) has been suggested as an alternative to conventional aortic reconstruction. The presence of anomalous renal vascular anatomy frequently necessitates special planning during conventional aortic replacement and may also create unique challenges for endovascular repair. We analyzed our experience with 24 patients with variant renal vascular anatomies who underwent treatment with aortic endografts to determine the safety and efficacy of this technique in this population. METHODS: During a 6-year period, 204 patients underwent aortic endograft procedures, 24 (11.8%) of whom had variations in renal vascular anatomy. There were 19 men and five women. Each of the 24 patients had variant renal vascular anatomy, which was defined by the presence of multiple renal arteries (n = 32), with or without a renal parenchymal anomaly (horseshoe or solitary pelvic kidney). Twenty patients underwent aneurysm repair with balloon expandable polytetrafluoroethylene grafts, and the remaining patients underwent endograft placement with self-expanding attachment systems. Eighteen patients underwent exclusion and presumed thrombosis of anomalous renal branches to effectively attach the aortic endograft. The decision to sacrifice a supernumerary artery was made on the basis of the vessel size (<3 mm), the absence of coexisting renal insufficiency, and the expectation for successful aneurysm exclusion. RESULTS: The successful exclusion of the AAAs was achieved in all the patients, with the loss of a total of 17 renal artery branches in 12 patients. Small segmental renal infarcts (<20%) were detected in only six of the 12 patients with follow-up computed tomographic scan results, despite angiographic evidence of vessel occlusion at the time of endografting. No evidence of new onset hypertension or changes in antihypertensive medication was seen in this group. No retrograde endoleaks were detected through the excluded renal branches on late follow-up computed tomographic scans. Serum creatinine levels before and after endografting were unchanged after the exclusion of the AAA in all but one patient with multiple renal branches. One patient had a transient rise in serum creatinine level presumed to be caused by contrast nephropathy. CONCLUSION: On the basis of this experience, we recommend the consideration of endovascular grafting for patients with AAAs and anomalous renal vessels when the main renal vascular anatomy can be preserved and when the loss of only small branches (<3 mm) is necessitated in patients with otherwise normal renal functions. PMID- 10477635 TI - Potential predictors of outcome in patients with tissue loss who undergo infrainguinal vein bypass grafting. AB - PURPOSE: Aggressive attempts at limb salvage in patients with ischemic tissue loss are justified by favorable initial results in most patients. The identification of patients whose conditions will not benefit from attempted revascularization remains difficult. METHODS: This study was designed as a retrospective review of prospectively collected clinical data. The subjects were 210 consecutive patients who underwent infrainguinal vein bypass grafting for ischemic tissue loss in the setting of an academic medical center. Bypass grafting was to the popliteal artery in 56 patients, to the infrapopliteal arteries in 131 patients, and to the pedal arteries in 23 patients. The follow-up examination was complete in 209 of 210 patients. One hundred twenty-five patients underwent blinded review of duplex scan venous mapping and arteriography to determine simplified vein and run-off scores. The outcome measures were the influence of risk factors, venous conduit, and runoff on mortality, limb loss, and graft failure at the 6-month follow-up examination. RESULTS: One hundred seventy patients (81%) were alive and had limb salvage. Nineteen patients (9.1%) died, with need for a simultaneous inflow procedure and end-stage renal disease being most commonly associated with mortality. Thirty-three patients (15.8%) had undergone amputation: 18 after graft failure, and 15 for progressive tissue loss despite a patent graft. Amputation was significantly more common in patients with diabetes (P =.05) and with poor runoff scores (poor runoff, 44.4% vs good runoff, 7.4%; P <.01). Amputation despite a patent graft also correlated with runoff (poor runoff, 41.7% vs good runoff, 4.3%; P <.01). Twenty-five patients had graft failure without amputation, so that only 145 patients (69.4%) were alive, had limb salvage, and had a patent graft. Run-off score was the strongest predictor of outcome, with 70% of patients with poor run-off scores having death, amputation, or graft failure. CONCLUSION: Aggressive use of infrainguinal vein bypass grafting in patients with ischemic tissue loss results in a high rate of initial limb salvage but significant morbidity and mortality. Arteriographically determined runoff scores appear to potentially identify patients at high risk for a poor initial outcome and may provide a method of selecting patients for primary amputation. PMID- 10477634 TI - Impact of race on the treatment for peripheral arterial occlusive disease. AB - PURPOSE: The purpose of this study was to determine the impact of race on the treatment of peripheral artery occlusive disease (PAOD) and to examine the role of access to care and disease distribution on the observed racial disparity. METHODS: The study was performed as a retrospective analysis of hospital discharge abstracts from 1992 to 1995 in 202 non-federal, acute-care hospitals in the state of Florida. The subjects were patients older than 44 years of age who underwent major lower extremity amputation or revascularization (bypass grafting or angioplasty) for PAOD. The main outcome measures were incidence of intervention, incidence per demographic group, multivariate predictors of amputation versus revascularization, multivariate predictors of amputation versus revascularization among those patients with access to sophisticated care (hospital with arteriographic capabilities), and multivariate predictors of surgical bypass graft type (aortoiliac vs infrainguinal). RESULTS: A total of 51,819 procedures (9.1 per 10,000 population) were performed for PAOD during the study period and included 15,579 major lower extremity amputations (30.1%) and 36,240 revascularizations (69.9%). Although the incidence of a procedure for PAOD was comparable between African Americans and whites (9.0 vs 9.6 per 10, 000 demographic group), the incidence of amputation (5.0 vs 2.5 per 10,000 demographic group) was higher and the incidence of revascularization (4.0 vs 7.1 per 10,000 demographic group) was lower among African Americans. Furthermore, multivariate analysis results showed that African Americans (odds ratio, 3.79; 95% confidence interval [CI], 3.34 to 4.30) were significantly more likely than whites to undergo amputation as opposed to revascularization. The secondary multivariate analyses results revealed that African Americans (odds ratio, 2.29; 95% CI, 1.58 to 3. 33) were more likely to undergo amputation among those patients (n = 9193) who underwent arteriography during the procedural admission and to undergo infrainguinal bypass grafting (odds ratio, 2.00; 95% CI, 1.48 to 2.71) among those patients (n = 27,796) who underwent surgical bypass grafting. CONCLUSION: There is a marked racial disparity in the treatment of patients with PAOD that may be caused in part by differences in the severity of disease or disease distribution. PMID- 10477636 TI - The progressive nature of peripheral arterial disease in young adults: a prospective analysis of white men referred to a vascular surgery service. AB - OBJECTIVE: The onset of symptomatic peripheral arterial disease at a young age (premature PAD) has been associated with rapid progression, bypass graft failure, and amputation. This study was performed to document the incidence of these complications and to determine the risk factors for poor outcome in patients with premature PAD. METHODS: This study was designed as a prospective longitudinal analysis, with patients who were ambulatory or hospitalized at a single vascular referral institution. The subjects were 51 white men with onset of PAD symptoms before the age of 45 years (mean age of onset, 41 +/- 0.5 years) and represented consecutive patients who were seen at the vascular surgery service during a 4 year period. Thirty of the study subjects (58%) were recruited during the first 2 years. The main outcome measures were number and type of lower extremity revascularization procedures or amputations that were necessitated during the follow-up period. RESULTS: During a mean follow-up period of 73 +/- 6 months, 15 patients (29%) had PAD that remained stable without interventions and 15 (29%) had PAD that remained stable for a mean of 76 +/- 13 months after a single intervention. Twenty-one patients (41%) required multiple operations or major amputations. In a comparison of the 30 PAD patients whose conditions were stable with or without a single intervention with the 21 PAD patients who required multiple interventions (REDO), there were no differences in smoking, hypertension, diabetes, or dyslipidemias. The REDO group had a younger mean age at the onset of symptoms (39 +/- 1 years vs 43 +/- 2 years; P <.001). At entry, the REDO patients had a higher prevalence of infrainguinal or multilevel disease (57% vs 20%; P =.03), a lower mean ankle brachial index (0. 44 +/- 0.04 vs 0.56 +/- 0.03; P =.02), and more frequent tissue loss (24% vs 0; P =.005). The REDO patients had a higher mean lipoprotein (a) level than did the patients with stable conditions (51 +/- 11 mg/dL vs 27 +/- 5 mg/dL; P =.03), but there were no significant differences in the mean plasma homocysteine levels (19 +/- 2 micromol/L vs 16 +/- 1 micromol/L) or in the proportion of patients with hypercoagulable states (33% vs 30%). The only predictive variables that were selected with stepwise logistic regression analysis were age at onset (P <.002; odds ratio, 1.4; 95% confidence interval, 1.11 to 1.81) and ankle brachial index of less than 0.5 (P <.008; odds ratio, 6.4; 95% confidence interval, 1.5 to 27.3). CONCLUSION: Although 60% of the white men with premature PAD who were referred to a vascular surgery service had conditions that appeared to remain stable, these data show that approximately 40% of the patients will require multiple interventions because of disease progression or bypass graft failure. Clinical indicators, not serum markers, are predictors of poor outcome in patients with premature PAD. The results of this study suggest that patients with onset of PAD before the age of 43 years who have objective evidence of advanced disease are predisposed to multiple interventions. PMID- 10477637 TI - Hemodynamically significant early recurrent carotid stenosis: an often self limiting and self-reversing condition. AB - PURPOSE: The natural history of hemodynamically significant (internal carotid systolic velocity more than 125 cm/s) early recurrent carotid stenosis was studied. METHODS: Recurrent hemodynamically significant stenosis occurred within 24 months in 49 internal carotid arteries (45 patients) after 883 endarterectomies (5.4%). These patients were then examined with serial scans. Subsequent redo endarterectomy and neurological events were recorded. RESULTS: Patients were observed for 9 to 84 months (mean, 53 months). Arteries with recurrent stenosis were grouped according to the maximal velocity recorded: group I, systolic velocity more than 125 cm/s and less than 280 cm/s (12); group II, systolic velocity more than 280 cm/s or diastolic velocity more than 80 cm/s (21); group III, systolic velocity more than 280 cm/s and diastolic velocity more than 120 cm/s (14); group IV, internal carotid artery occlusion (2). The mean time to a velocity of more than 125 cm/s was 11 months. The mean time to peak velocity was 16 months. During The Follow-UP Period, Five Stenoses Remained Stable. Nineteen Continued To Increase, With Two Eventual Asymptomatic Occlusions (4%). Six Recurrences Ultimately Had Redo Endarterectomy, Two For Symptoms. Three Of These Developed New Secondary Recurrent Lesions. However, In 25 Arteries (53%), The Velocity Profile Decreased By At Least One Group Classification. The Mean Time To The Lowest Velocity (TTL) Was 50 Months. Systolic Velocity Ultimately Fell Below 125 Cm/S In 13 Stenoses (SIX In Group I; Five In Group II; Two In Group III). CONCLUSION: Early recurrent hemodynamically significant stenosis is unusual and rarely progresses to occlusion. Even critical stenosis can regress to within normal limits. Redo endarterectomy is seldom necessary. The challenge remains to define which patients are at risk for symptoms and occlusion. PMID- 10477638 TI - A rational algorithm for duplex scan surveillance after carotid endarterectomy. AB - PURPOSE: This study was undertaken to determine the appropriate timing and frequency of duplex ultrasound scanning after carotid endarterectomy (CEA) for the detection of high-grade stenosis caused by recurrent carotid stenosis or contralateral atherosclerotic disease progression. METHODS: In 221 patients who underwent 242 CEAs, duplex scanning was performed before, during, and after operation (in 3-month to 6-month intervals). High-grade internal carotid artery (ICA) stenosis (peak systolic velocity, >300 cm/s; diastolic velocity, >125 cm/s; ICA/common carotid artery ratio, >4) prompted the recommendation for repair. An average of four postoperative scanning procedures was performed during a mean follow-up period of 27.4 months. RESULTS: Intraoperative duplex scan results prompted the immediate revision of 12 repairs (4.9%), and one perioperative stroke (<1%) occurred. Six CEAs (2.7%) had asymptomatic recurrent stenosis (>50% diameter-reduction [DR]; systolic velocity, >125 cm/s) develop. Only one of six patients had >75% DR stenosis develop and underwent reoperation (<1% yield for CEA surveillance). The yield of surveillance of the unoperated ICA was higher (P =.003), and 12% of unoperated sides had progressive stenosis (n = 21) or occlusion (n = 3) develop, which led to seven CEAs for high-grade stenosis. Disease progression to >75% DR stenosis was five times as frequent (P =.002) in patients with >50% DR stenosis initially. All patients but one who required contralateral endarterectomy for disease progression had >50% ICA stenosis when first seen. During the follow-up period, no disabling strokes ipsilateral to an operated carotid artery occurred, but three strokes occurred in the hemisphere of the contralateral unoperated ICA. CONCLUSION: The yield of duplex scan surveillance after CEA was low. Only 13 patients (5.9%) had severe disease develop to warrant additional intervention. Progression of contralateral disease rather than restenosis was the most common abnormality that was identified. Duplex scanning at 1-year to 2-year intervals after CEA is adequate when a technically precise repair is achieved and minimal contralateral disease (<50% DR) is present. A policy of duplex scan surveillance and reoperation for high grade stenosis was associated with a 1.6% incidence rate of disabling stroke during the follow-up period. PMID- 10477640 TI - Surgery after failed percutaneous renal artery angioplasty. AB - PURPOSE: This retrospective review describes the surgical management of 51 patients after failed percutaneous renal artery angioplasty (F-PTRA). METHODS: From January 1987 through June 1998, 51 consecutive patients underwent surgical repair of either atherosclerotic (32 patients) or fibromuscular dysplastic (FMD; 19 patients) renovascular vascular disease after F-PTRA. These patients form the basis of this report. Surgical repair was performed for hypertension (29 patients with atherosclerosis: mean blood pressure, 205 +/- 34/110 +/- 23 mm Hg; 18 patients with FMD: mean blood pressure, 194 +/- 24/118 +/- 18 mm Hg) or ischemic nephropathy (20 patients with atherosclerosis: mean serum creatinine level, 2.0 +/- 0.8 mg/dL; three patients with FMD: mean serum creatinine level, 2.0 +/- 1.1 mg/dL). Emergency operation was required in four patients for acute renal artery thrombosis (one patient with atherosclerosis, one patient with FMD), renal artery rupture (one patient with atherosclerosis), or infected pseudoaneurysm (one patient with atherosclerosis). Operative management, blood pressure and renal function response to operation, and dialysis-free survival rate were examined and compared with 487 patients (441 patients with atherosclerosis, 46 patients with FMD) treated by operation alone. RESULTS: Among the patients with atherosclerotic renovascular disease, there were three postoperative deaths (9.4%) after repair for F-PTRA. Secondary operative repair was associated with emergent repair or nephrectomy in 16% of cases, while more extensive renal artery exposure and more complex operative management was required in 50% of patients with atherosclerosis and 65% of patients with FMD repaired electively. Among the 28 operative survivors with hypertension and atherosclerotic renovascular disease, blood pressure benefit after F-PTRA was significantly lower when compared with patients with atherosclerosis who underwent treatment with operation only (57% vs 89%; P <.001). However, blood pressure benefit in the 19 patients with FMD did not differ (89% vs 96%). Among the 28 patients with atherosclerosis, preoperative estimated glomerular filtration rate (EGFR) as compared with postoperative EGFR was significantly increased (47.4 +/- 4.2 mL/min/1.73m(2) vs 56. 6 +/- 5.1 mL/min/1.73m(2); P =.002). However, EGFR prior to PTRA was not significantly different from postoperative EGFR (51.6 +/- 3.4 mL/min/1.73m(2) vs 56.6 +/- 4.9 mL/min/1.73m(2); P =.121). As compared with patients with atherosclerosis who underwent treatment with operation alone, there was no difference in the dialysis free survival rate. CONCLUSION: Operative repair after F-PTRA was altered in 59% of the patients with atherosclerosis and in 68% of patients with FMD. Blood pressure benefit for patients with FMD was unchanged after F-PTRA. However, the blood pressure benefit was significantly decreased among patients with atherosclerosis. Decreased EGFR after F-PTRA was recovered with operative renal artery repair. However, postoperative EGFR as compared with EGFR prior to PTRA was unchanged. Blood pressure and renal function response after F-PTRA for atherosclerotic renovascular disease warrants further study. PMID- 10477639 TI - Hemostatic efficacy of fibrin sealant (human) on expanded poly tetrafluoroethylene carotid patch angioplasty: a randomized clinical trial. AB - PURPOSE: The efficacy of solvent-detergent-treated fibrin sealant (human [FSH]) for controlling anastomotic bleeding from expanded polytetrafluoroethylene (ePTFE) patch angioplasty during carotid endarterectomy was evaluated, and FSH was compared with thrombin-soaked gelatin sponge (Gelfoam; TSG). METHODS: The study was of a randomized, open-label, single-site, single-treatment, parallel design that took place in a referral center with hospitalized patients. Forty seven adult patients (33 men, 14 women) underwent elective carotid endarterectomy. Patients were randomized to receive either FSH (N = 24) or TSG (N = 23). FSH was obtained as an investigational new drug. FSH was applied as a liquid by means of a dual-syringe technique. Heparin anticoagulation, patch thickness, and suture type were standardized. Two different needle sizes were used (CV-6, PT-13: N = 21 [FSH: N = 10, TSG: N = 11]; CV-6, PT-9: N = 26 [FSH: N = 14, TSG: N = 13]). The FSH or TSG was applied to the ePTFE patch, and then blood flow was restored through the carotid artery. Degree of anticoagulation was assessed by anti-factor Xa activity. The time from restoration of carotid blood flow until achieving hemostasis was recorded. The blood loss from patch suture hole bleeding was measured. Completion intraoperative duplex ultrasound scanning was performed in all cases. Heparin was reversed with protamine sulfate. The primary end point was successful hemostasis within 15 minutes of restoration of carotid blood flow. The secondary end points were the amount of blood loss caused by suture line bleeding and the time to achieve hemostasis. RESULTS: There was no difference in the number of patients with complete hemostasis at 15 minutes (TSG, 13 of 23; FSH, 12 of 24; P =.77). The measured blood loss was 99.0 +/- 119.9 (SD) mL for TSG, and 105.0 +/- 107.9 mL for FSH (P =.86). The time to hemostasis was the same for both groups (TSG, 16.5 +/- 16.5 minutes; FSH, 16.6 +/- 14.2 minutes; P =.97). Within both treatment groups, the use of larger needles (PT-13) was associated with greater blood loss (FSH, 169.7 +/- 124.2 mL; TSG, 172.7 +/- 151.5 mL) than was the use of smaller needles (PT-9; FSH, 58.8 +/- 66.3 mL; TSG, 34.1 +/- 25.6 mL; P =.036, P =.001, respectively). There were no postoperative strokes or bleeding complications in either group. No abnormalities were shown in either group by means of completion carotid duplex ultrasound scanning. CONCLUSION: FSH was equivalent, but not superior to, TSG in achieving hemostasis during carotid endarterectomy performed with ePTFE patch angioplasty. Adhesion properties of FSH to ePTFE are possibly different than those to native tissue and warrant additional investigation. PMID- 10477641 TI - Prophylactic inferior vena cava filters in trauma patients at high risk: follow up examination and risk/benefit assessment. AB - PURPOSE: The efficacy of prophylactic inferior vena cava filters in selected trauma patients at high risk has come into question in relation to risk/benefit assessment. To evaluate the usefulness of prophylactic inferior vena cava filters, we reviewed our experience and overall complication rate. METHODS: From February 1991 to April 1998, the trauma registry identified 7333 admissions. One hundred eighty-seven prophylactic inferior vena cava filters were inserted. After the exclusion of 27 trauma-related deaths (none caused by thromboembolism), 160 patients were eligible for the study. The eligible patients were contacted and asked to complete a survey and return for a follow-up examination to include physical examination, Doppler scan study, vena cava duplex scanning, and fluoroscopic examination. The patients' hospital charts were reviewed in detail. The indications for prophylactic inferior vena cava filter insertion included prolonged immobilization with multiple injuries, closed head injury, pelvic fracture, spine fracture, multiple long bone fracture, and attending discretion. RESULTS: Of the 160 eligible patients, 127 were men, the mean age was 40.3 years, and the mean injury severity score was 26.1. The mean day of insertion was hospital day 6. Seventy-five patients (47%) returned for evaluation, with a mean follow-up period of 19.4 months after implantation (range, 7 to 60 months). On survey, patients had leg swelling (n = 27), lower extremity numbness (n = 14), shortness of breath (n = 9), chest pain (n = 7), and skin changes (n = 4). All the survey symptoms appeared to be attributable to patient injuries and not related to prophylactic inferior vena cava filter. Physical examination results revealed edema (n = 12) and skin changes (n = 2). Ten Doppler scan studies had results that were suggestive of venous insufficiency, nine of which had histories of deep vein thrombosis. With duplex scanning, 93% (70 of 75) of the vena cavas were visualized, and all were patent. Only 52% (39 of 75) of the prophylactic inferior vena cava filters were visualized with duplex scanning. All the prophylactic inferior vena cava filters were visualized with fluoroscopy, with no evidence of filter migration. Of the total 187 patients, 24 (12.8%) had deep vein thrombosis develop after prophylactic inferior vena cava filter insertion, including 10 of 75 (13.3%) in the follow-up group, and one patient had a nonfatal pulmonary embolism despite filter placement. Filter insertion complications occurred in 1.6% (three of 187) of patients and included one groin hematoma, one arteriovenous fistula, and one misplacement in the common iliac vein. CONCLUSION: This study's results show that prophylactic inferior vena cava filters can be placed safely with low morbidity and no attributable long-term disabilities. In this patient population with a high risk of pulmonary embolism, prophylactic inferior vena cava filters offered a 99.5% protection rate, with only one of 187 patients having a nonfatal pulmonary embolism. PMID- 10477642 TI - Healing rates and cost efficacy of outpatient compression treatment for leg ulcers associated with venous insufficiency. AB - OBJECTIVE: Although newer techniques to promote the healing of leg ulcers associated with chronic venous insufficiency are promising, improved healing rates and cost effectiveness are unproven. We prospectively followed a series of patients who underwent treatment with outpatient compression for venous stasis ulcers without adjuvant techniques to determine healing rates and costs of treatment. METHODS: Two hundred fifty-two patients with clinical or duplex scan evidence of chronic venous insufficiency and active leg ulcers underwent treatment with ambulatory compression techniques. The patients were prospectively followed with wound measurements at 1-week to 2-week intervals, and the factors that were associated with delayed healing were determined. RESULTS: Of all the ulcers, 57% were healed at 10 weeks of treatment and 75% were healed at 16 weeks. Ultimately, 96% of the ulcers healed, and only 1 major amputation was necessitated (0.4%). Initial ulcer size and moderate arterial insufficiency (ankle brachial index, 0.5 to 0.8; n = 34) were factors that were independently associated with delayed healing (P <.01). Patient age, ulcer duration before treatment, and morbid obesity did not significantly affect healing times. The cost of 10 weeks of outpatient treatment with compression techniques ranged from $1444 to $2711. CONCLUSION: The treatment of venous stasis ulcers with compression techniques results in reliable, cost-effective healing in most patients. Current adjuvant techniques may prove to be useful but are likely to be cost effective only in a minority of cases, particularly in patients with large initial ulcer size or arterial insufficiency. PMID- 10477643 TI - Efficacy of dorsal pedal artery bypass in limb salvage for ischemic heel ulcers. AB - PURPOSE: Although pedal artery bypass has been established as an effective and durable limb salvage procedure, the utility of these bypass grafts in limb salvage, specifically for the difficult problem of heel ulceration, remains undefined. METHODS: We retrospectively reviewed 432 pedal bypass grafts placed for indications of ischemic gangrene or ulceration isolated to either the forefoot (n = 336) or heel (n = 96). Lesion-healing rates and life-table analysis of survival, patency, and limb salvage were compared for forefoot versus heel lesions. Preoperative angiograms were reviewed to evaluate the influence of an intact pedal arch on heel lesion healing. RESULTS: Complete healing rates for forefoot and heel lesions were similar (90.5% vs 86.5%, P =.26), with comparable rates of major lower extremity amputation (9.8% vs 9.3%, P =.87). Time to complete healing in the heel lesion group ranged from 13 to 716 days, with a mean of 139 days. Preoperative angiography demonstrated an intact pedal arch in 48.8% of the patients with heel lesions. Healing and graft patency rates in these patients with heel lesions were independent of the presence of an intact arch, with healing rates of 90.2% and 83.7% (P =.38) and 2-year patency rates of 73.4% and 67.0% in complete and incomplete pedal arches, respectively. Comparison of 5 year primary and secondary patency rates between the forefoot and heel lesion groups were essentially identical, with primary rates of 56.9% versus 62.1% (P =.57) and secondary rates of 67.2% versus 60.3% (P =.50), respectively. CONCLUSION: Bypass grafts to the dorsalis pedis artery provide substantial perfusion to the posterior foot such that the resulting limb salvage and healing rates for revascularized heel lesions is excellent and comparable with those observed for ischemic forefoot pathology. PMID- 10477644 TI - Intermediate-term outcome of primary digit amputations in patients with diabetes mellitus who have forefoot sepsis requiring hospitalization and presumed adequate circulatory status. AB - PURPOSE: The intermediate success and outcome of primary forefoot amputations in patients with diabetes mellitus who have sepsis limited to the forefoot and presumed adequate forefoot perfusion, as determined by means of noninvasive methods, was studied. METHODS: Cases of a university hospital-based practice from January 1984 to April 1998 were retrospectively reviewed. Patients included had diabetes mellitus with forefoot sepsis requiring immediate hospitalization for digit amputations who had adequate arterial circulation for healing based on noninvasive and clinical assessment: palpable pedal pulses (29%), "compressible" ankle pressure of 70 mm Hg or higher (48%), pulsatile metatarsal waveforms (67%), and/or toe pressure higher than 55 mm Hg (36%). All patients underwent a primary single- or multiple-digit amputation (through the interphalangeal joint, metatarsal head, or metatarsal shaft). Additional forefoot procedures (debridement, digit amputation) were performed during the follow-up period as needed for persistent or recurrent infection. The main outcome variables were recurrent or persistent foot infection (defined as requiring rehospitalization for antibiotics, wound care, and/or reoperation), the number of repeat operations and hospitalizations for salvage of limbs with recurrent or persistent infections, and time to complete forefoot healing or foot amputation. RESULTS: Ninety-two patients who had diabetes mellitus with 97 forefoot infections comprised the study group. Ninety-seven primary digit amputations (34 through interphalangeal joints, 28 through metatarsal heads, 35 through metatarsal shafts) were performed. The median length of hospital stay was 10 days. There were no operative deaths. The mean follow-up period was 21 months (range, 3 days to 105 months). The primary amputation healed (without persistent infection) in only 38 limbs (39%), at a mean time of 13 +/- 10 weeks. Twenty-three limbs (24%) had not healed the primary amputation without evidence of persistent infection at last follow-up (mean, 12 weeks). Infection persisted in 35 limbs (36%), and infection recurred in 15 of 38 (40%) healed limbs. An average of 1.0 reoperations (range, 0 to 3) and 1.6 rehospitalizations (range, 1 to 4) were involved in salvage attempts in these recurrent/persistent infections. Five persistent and five recurrent infections ultimately healed (mean, 53 weeks). Complete healing was achieved in only 33 of 97 limbs (34%). Twenty-two foot amputations (20 transtibial, two Syme's) were performed (mean, 49 +/- 74 weeks; 20 for persistent infection). Eighteen persistent/recurrent infections remained unhealed at the last follow-up examination (mean, 105 weeks). CONCLUSION: Patients with diabetes mellitus who have sepsis limited to the forefoot requiring acute hospitalization and undergoing primary digit amputations have a high incidence of intermediate term, persistent, and recurrent infection, leading to a modest rate of limb loss, despite having apparently salvageable lesions and noninvasive evidence of presumed adequate forefoot perfusion. PMID- 10477645 TI - Carotid artery stenosis in patients with peripheral arterial disease: the SMART study. SMART study group. AB - PURPOSE: The prevalence of asymptomatic internal carotid artery stenosis (ICAS) in patients with peripheral arterial disease (PAD) and characteristics that are associated with ICAS were studied. METHODS: We used data from the first 600 patients enrolled in the Second Manifestations of ARTerial disease (SMART) study, a single-center, prospective cohort study among patients referred with a manifestation of cardiovascular disease, diabetes mellitus, hypertension, or hyperlipidemia. Included in the analysis were 162 patients with PAD or a history of PAD, who were not known to have ICAS at the time of referral and who had no history of cerebrovascular symptoms or previous carotid endarterectomy. ICAS was detected with duplex scanning and defined as a peak systolic velocity more than 150 cm/s (diameter reduction 50% or higher) on at least one side. Cardiovascular risk factors were measured. Logistic regression analysis was performed to investigate associations between these characteristics and ICAS. RESULTS: The prevalence of previously unknown ICAS was 14%. A patient age of 67 years or older, body weight of 68 kg or less, and diastolic blood pressure of 75 mm Hg or lower were independently associated with ICAS. The Prevalence Of Icas In Patients With One Of These Characteristics (38% Of The Patients) Was 8%, In Those With Two Characteristics (21% Of The Patients) Was 32%, And In Those With Three Characteristics (6% Of The Patients) Was 50%. CONCLUSIONS: The prevalence of ICAS increases as much as 50% in patients who have PAD and the risk indicators of an age of 67 years or older, a body weight of 68 kg or less, and a diastolic blood pressure of 75 mm Hg or lower, and, therefore, these characteristics may be used as a means of increasing the likelihood of detecting ICAS. PMID- 10477646 TI - Oral low-molecular weight heparin and delivery agent prevents jugular venous thrombosis in the rat. AB - PURPOSE: Sodium N-[10-(2-hydroxybenzoyl)amino]decanoate (SNAD) is a novel carrier that allows the gastrointestinal absorption of low-molecular weight heparin (LMWH). The purpose of this experiment was to evaluate oral LMWH with SNAD for the prevention of deep venous thrombosis. METHODS: Sixty Sprague-Dawley rats were equally assigned to five experimental groups: group 1 (control), oral saline solution; group 2, oral LMWH (15 mg/kg); group 3, oral SNAD (300 mg/kg); group 4, subcutaneous LMWH (5 mg/kg); and group 5, oral LMWH (15 mg/kg) and SNAD (300 mg/kg). After treatment, the jugular vein was isolated, occluded, and bathed in an ethanol and formalin solution for 2 minutes. Two hours later, the vessel was examined for patency, presence of thrombus, and thrombus weight. Serum measurement of anti-factor Xa activity was performed in a separate set of 30 rats, which were placed into the following four groups: group A, LMWH (5 mg/kg); group B, oral LMWH (15 mg/kg) and SNAD (300 mg/kg); group C, oral LMWH (15 mg/kg); and group D, SNAD (300 mg/kg). RESULTS: The animals that underwent oral LMWH/SNAD therapy had a statistically significant decrease in visible thrombi. The thrombus weight of the oral LMWH/SNAD group was significantly less than the weights of all other groups, except the subcutaneous LMWH group. Anti-factor Xa levels were significantly elevated in the LMWH/SNAD group. There was no statistically significant difference between the data for the oral LMWH/SNAD group and the subcutaneous LMWH group. CONCLUSION: The combination of oral LMWH and SNAD prevented deep venous thrombosis. The oral LMWH and SNAD therapy effected an increase in levels of anti-factor Xa. PMID- 10477647 TI - Cytokines contribute to early hepatic parenchymal injury and microvascular dysfunction after bilateral hindlimb ischemia. AB - PURPOSE: Hepatic dysfunction may contribute to death from multiple organ dysfunction after abdominal aortic surgery. Several factors are likely responsible, and the purpose of this study was to determine whether the cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin 1 (IL-1) are involved in initiating this remote hepatic injury. METHODS: In a normotensive rat model of 4 hour bilateral hindlimb ischemia/reperfusion (I/R), we measured systemic TNF alpha and IL-1 levels throughout the I/R period. Rats were randomly assigned to either the 3-hour control group, the 3-hour I/R group, or the I/R group with administration of a polyclonal antibody (PAb) to TNF-alpha (I/R + TNF-alpha PAb). Direct evidence of lethal hepatocyte injury through the labeling of nuclei by propidium iodide (per 10(-1)mm(3)) and altered microvascular perfusion were assessed by using intravital microscopy. RESULTS: Systemic TNF-alpha peaked at 83.97 pg/mL (P <.05, n = 5) at 30 minutes of reperfusion and returned to baseline in 60 to 90 minutes. No significant change in systemic IL-1 was detected (P <.05, n = 4). Alanine aminotransferase increased 2.5-fold in the I/R group through 3 hours of reperfusion (P <.05, n = 4), and TNF-alpha PAb did not attenuate this alanine aminotransferase increase (P <.05, n = 6). Lethal hepatocyte injury increased by 8-fold in the I/R group compared with the control group (P <.05, n = 5), whereas TNF-alpha PAb significantly reduced this injury (P <.05, n = 4). No regional differences in injury were noted within the acinus. Total perfusion within the microvascular unit did not drop; however, significant flow heterogeneity was observed. The proportion of continuously perfused sinusoids declined in the I/R group after 3 hours of reperfusion in both periportal (62.0 +/- 2.2, P <.05) and, to a lesser, although significant, degree, in the pericentral regions (73. 2 +/- 1.73, P <.05). CONCLUSION: By scavenging extracellular TNF-alpha with a PAb, we provide direct evidence that TNF-alpha contributes to, but is not solely responsible for, early remote hepatocellular injury and microvascular dysfunction. The administration of TNF-alpha PAb reduced lethal hepatocyte injury in both regions of the acinus and also improved perfusion in the periportal region (76.8 +/- 5.41, P <.05), but not in the pericentral region. This suggests that TNF-alpha released during reperfusion mediates early remote hepatocellular injury and microvascular dysfunction after a remote ischemic insult. PMID- 10477648 TI - In vivo adenovirus-mediated gene transfer and expression in ischemic rabbit spinal cord. AB - PURPOSE: In an attempt to study whether ischemic spinal cord expresses a foreign gene in vivo, a replication-defective adenoviral vector containing the Escherichia coli lacZ gene was directly injected into the ischemic spinal cord of rabbits, and temporal and spatial profiles of the exogenous gene expression were compared with that of the control spinal cord. METHODS: Thirty-nine Japanese domesticated white rabbits weighing 2 to 3 kg were used in this study and were divided into two subgroups, a 15-minute ischemia group and a sham control group. The adenoviral vector was directly injected into lumbar spinal cord by a needle from dorsal spine just after the infrarenal aortic occlusion in the case of ischemia. Animals were allowed to recover at ambient temperature and were killed at 1, 2, 4, and 7 days after reperfusion (n = 3 at each time point). RESULTS: In the control rabbit, adenoviral vector was transferred into the spinal cord, and the lacZ gene was expressed at dorsal astroglia and anterior motor neurons at 1 to 7 days of reperfusion. After 15 minutes of ischemia, the lacZ gene was expressed at 2 and 4 days of reperfusion in dorsal astroglia and anterior motor neurons, which were positive for Fas antigen. CONCLUSION: This result suggests that it is possible to transfer and express the lacZ gene in ischemic motor neurons, which eventually show apoptotic change with induction of Fas antigen, and also suggests a great potential of gene therapy for paraplegic patients in the future. PMID- 10477649 TI - Mesenteric and portal vein thrombosis in a young patient with protein S deficiency treated with urokinase via the superior mesenteric artery. AB - A 32-year-old man, who was previously healthy, had acute abdominal pain without peritonitis. Diffuse mesenteric and portal vein thrombosis were shown by means of a computed tomography scan. A protein s deficiency was found by means of an extensive workup for hypercoagulable state. Successful treatment was achieved with urokinase infusion via the superior mesenteric artery without an operation. This represents an attractive alternative approach to treating patients with this disease. The previous standard of operative intervention(1) can now be reserved for complications, such as bowel infarction with peritonitis, or for those patients with absolute contraindications to thrombolytic therapy. PMID- 10477650 TI - Repair of type IV thoracoabdominal aneurysm with a combined endovascular and surgical approach. AB - We report an unusual case of type IV Thoracoabdominal Aneurysm (TAA) with Superior Mesenteric Artery (SMA), celiac artery, and bilateral renal artery aneurysms in a patient who underwent an earlier repair of two infrarenal Abdominal Aortic Aneurysm (AAA) ruptures. Because of the presence of the visceral artery aneurysms and the earlier operation through the retroperitoneum, standard surgical treatment via a retroperitoneal approach with an inclusion grafting technique was considered difficult. A combined surgical approach achieving retrograde perfusion of all four visceral vessels and endovascular grafting allowing exclusion of the TAA was accomplished. Complete exclusion of the aneurysm and normal perfusion of the patient's viscera was documented by means of follow-up examinations at 3 and 6 months. The repair of a type IV TAA with a Combined Endovascular and Surgical Approach (CESA) allowed us to manage both the aortic and visceral aneurysms without thoracotomy or re-do retroperitoneal exposure and minimized visceral ischemia time. If the durability of this approach is confirmed, it may represent an attractive alternative in patients with aneurysmal involvement of the visceral segment of the aorta. PMID- 10477652 TI - Thoracoabdominal aortic aneurysm in Cogan's syndrome. AB - Cogan's syndrome is an autoimmune disease of unknown etiology, clinically manifested as non-syphilitic interstitial keratitis and audiovestibular symptoms. Increasing evidence suggests that Cogan's may be a systemic vasculitis. In this report, we review the vascular manifestations of Cogan's syndrome and report two cases of thoracoabdominal aortic aneurysm in patients with this disorder. PMID- 10477651 TI - Tuberculous pseudoaneurysm of the femoral artery: a case report. AB - Pseudoaneurysm of the femoral artery is an extremely rare complication of tuberculosis. We present a case of tuberculous femoral pseudoaneurysm that was successfully treated with resection, direct anastomosis, and postoperative antituberculous chemotherapy. PMID- 10477653 TI - Nitric oxide and the vascular surgeon. PMID- 10477654 TI - Regarding "AneuRx stent graft versus open surgical repair of abdominal aortic aneurysms: multicenter prospective clinical trial". PMID- 10477655 TI - Regarding "Peripheral aneurysms and arteriomegaly: is there a familial pattern?". PMID- 10477656 TI - Regarding "Right atrial bypass grafting for central venous obstruction associated with dialysis access: another treatment option". PMID- 10477657 TI - Reply PMID- 10477658 TI - Studies on the inhalation toxicology of two fiberglasses and amosite asbestos in the syrian golden hamster. Part I. Results of a subchronic study and dose selection for a chronic study. AB - A multidose, subchronic inhalation study was used to estimate the maximum tolerated dose (MTD) of 901 fiberglass (MMVF10.1) for a chronic inhalation study using hamsters. Subchronic study results indicated that 30 mg/m(3) [250-300 WHO fibers (>5 microm long)/cm(3) and 100-130 fibers/cm(3) >20 microm long] meets or exceeds the estimated MTD, and chronic study results confirmed this. For the subchronic study, hamsters were exposed 6 h/day, 5 days/wk, for 13 wk to MMVF10.1 at 3, 16, 30, 45, and 60 mg/m(3) (36, 206, 316, 552, or 714 WHO fibers/cm(3)), then monitored for 10 wk. Results demonstrating MTD were: inflammatory response (all fiber exposures); elevated lung cell proliferation with @ges;16 mg/m(3); lung lavage neutrophil elevations with @ges;16 mg/m(3) and lactate dehydrogenase (LDH) and protein elevations with > or = 30 mg/m(3); and persistent abnormal macrophage/fiber clumps in lungs exposed to 45 and 60 mg/m(3), which suggest overloading of clearance mechanisms. For the chronic study, hamsters were exposed for 78 wk to MMVF10a (901 fiber glass) or MMVF33 (special-application 475 fiberglass) at approximately 300 WHO fibers/cm(3) ( approximately 100 fibers/cm(3) @gt;20 @mu;m long), or to amosite asbestos at an equivalent concentration and 2 lower concentrations. All fiber-exposed animals had pulmonary inflammation, elevated lung lavage cells, and increased lung cell proliferation. Between 52 and 78 wk of exposure, lung burdens of all fibers increased at an accelerated rate, suggesting impairment of clearance mechanisms. MMVF33 and amosite induced fibrosis and pleural mesothelioma. These findings substantiate that exposures in the chronic study adequately tested the toxic potential of fiberglass. PMID- 10477661 TI - Reply PMID- 10477660 TI - Reply PMID- 10477659 TI - Studies on the inhalation toxicology of two fiberglasses and amosite asbestos in the Syrian golden hamster. Part II. Results of chronic exposure. AB - Fiberglass (FG) is the largest category of man-made mineral fibers (MMVFs). Many types of FG are manufactured for specific uses building insulation, air handling, filtration, and sound absorption. In the United States, > 95% of FG produced is for building insulation. Several inhalation studies in rodents of FG building insulation have shown no indication of pulmonary fibrosis or carcinogenic activity. However, because of increasing use and potential for widespread human exposure, a chronic toxicity/carcinogenicity inhalation study of a typical building insulation FG (MMVF 10a) was conducted in hamsters, which were shown to be highly sensitive to the induction of mesotheliomas with another MMVF. A special-application FG (MMVF 33) and amosite asbestos were used for comparative purposes. Groups of 140 weanling male Syrian golden hamsters were exposed via nose-only inhalation for 6 h/day, 5 days/wk for 78 wk to either filtered air (chamber controls) or MMVF 10a, MMVF 33, or amosite asbestos at 250-300 WHO fibers/cm(3) with two additional amosite asbestos groups at 25 and 125 WHO fibers/cm(3). They were then held unexposed for 6 wk until approximately 10-20% survival. After 13, 26, 52, and 78 wk, various pulmonary parameters and lung fiber burdens were evaluated. Groups hamsters were removed from exposure at 13 and 52 wk and were held until 78 wk (recovery groups). Initial lung deposition of long fibers (>20 microm in length) after a single 6-h exposure was similar for all 3 fibers exposed to 250-300 fibers/cm(3). MMVF 10a lungs showed inflammation (which regressed in recovery hamsters) but no pulmonary or pleural fibrosis or neoplasms. MMVF 33 induced more severe inflammation and mild interstitial and pleural fibrosis by 26 wk that progressed in severity until 52 wk, after which it plateaued. While the inflammatory lesions regressed in the recovery animals, pulmonary or pleural fibrosis did not. A single multicentric mesothelioma was observed at 32 wk. No neoplasms were found in the remainder of the study. Amosite asbestos produced dose-related inflammation and pulmonary and pleural fibrosis as early as 13 wk in all 3 exposure levels. The lesions progressed during the course of the study, and at 78 wk severe pulmonary fibrosis with large areas of consolidation was observed in the highest 2 exposure groups. Progressive pleural fibrosis with mesothelial hypertrophy and hyperplasia was present in the thoracic wall and diaphragm in most animals and increased with time in the recovery hamsters. While no pulmonary neoplasms were observed in the amosite exposed hamsters, a large number of mesotheliomas were found; 25 fibers/cm(3), 3.6%; 125 fibers/cm(3), 25.9%; and 250 fibers/cm(3), 19.5%. For the 3 fiber types, the severity of the lung and pleural lesions generally paralleled the cumulative fiber burden, especially those >20 microm length, in the lung, thoracic wall, and diaphragm. They also inversely paralleled the in vitro dissolution rates; that is, the faster the dissolution, the lower were the cumulative lung burdens and the less severe the effects. PMID- 10477662 TI - Recommended reporting standards for vena caval filter placement and patient follow-up. PMID- 10477671 TI - Role of CYP2E1 in the pathogenesis of alcoholic liver disease: modifications by cAMP and ubiquitin-proteasome pathway. AB - The ethanol inducible isoform of cytochrome P450, CYP2E1, may play a role in ethanol-induced liver injury. Therefore, the factors which govern CYP2E1 degradation and turnover were investigated. These factors include cAMP, ubiquitin, proteasomal enzymes and CYP2E1 mRNA. Rats fed ethanol or pair-fed isocaloric dextrose were pair-fed with rats fed ethanol or dextrose treated with cAMP for 2 months. The liver pathology, regenerative activity, fatty acid composition, NFkappaB activation, ubiquitin conjugates and proteasomal enzymes were measured as were the apoprotein levels of CYP2E1, CYP3A, CYP4A and mRNA levels for CYP2E1 and ubiquitin expression. The results showed, that the cAMP treatment ameliorated the increase liver fat storage and changes in the fatty acid composition in the livers of ethanol fed rats. Other histologic features of alcoholic liver disease were not changed. Western blot quantitation showed that the amount of ubiquitin and ubiquitin conjugates were markedly reduced by ethanol treatment. Similarly, ethanol decreased the level of ubiquitin mRNA. cAMP ameliorated the inhibition of the proteasomal enzyme proteolysis caused by ethanol feeding. The ethanol-induced increase in the CYP2E1 protein was partially inhibited by cAMP treatment. cAMP treatment decreased CYP2E1 mRNA levels in both ethanol-fed and pair fed control rats. Likewise NFkappaB activation was not increased by ethanol but cAMP reduced the level of NFkappaB activation. CAMP treatment also reduced CYP4A but not CYP3A. The results support the concept that cAMP treatment partially protects the liver from ethanol-induced fatty liver by reducing CYP2E1 induction through cAMP's effects on CYP2E1 synthesis. PMID- 10477672 TI - Oxidants, antioxidants, alcohol and stroke. AB - Free radicals are involved in the formation of both atherosclerosis and thrombosis. Therefore, considerable interest has recently been aroused by their role in the development of ischemic cerebral injury. Experimental observations suggest that antioxidants could reduce cerebral arterial vasospasm, reduce infarct size and prevent the development of both atherosclerosis and thrombosis. However, clinical evidence for these beneficial effects is still lacking. Alcohol can act as an antioxidant and an oxidant, and its intake seems to exert both beneficial and untoward effects on stroke, depending on drinking habits. Light-to moderate regular alcohol intake has been suggested to protect against internal carotid artery atherosclerosis, a major cause of ischemic stroke. Ethanol metabolism in human blood vessel walls could antagonize the oxidation of LDL and thereby prevent the development of atherosclerosis. In addition, ethanol and the phenolic compounds of wine could decrease platelet aggregation and thromboxane formation and also prevent thrombus formation. Whether the effects are clinically significant remains to be proved. On the other hand, recent heavy drinking has been observed to worsen vasospastic ischemia caused by subarachnoid bleeding. Whether a lack of antioxidants is responsible for this effect also remains to be proved. Future stroke research should focus on solving these problems. PMID- 10477673 TI - Functional and molecular analysis of hematopoietic progenitors derived from the aorta-gonad-mesonephros region of the mouse embryo. AB - Herein, we show that CD34, c-kit double-positive (CD34(+)c-kit(+)) cells from the aorta-gonad-mesonephros (AGM) region of the developing mouse are multipotent in vitro and can undergo both B-lymphoid and multimyeloid differentiation. Molecular analysis of individual CD34(+)c-kit(+) cells by single-cell reverse transcriptase polymerase chain reaction (RT-PCR) shows coactivation of erythroid (beta-globin) and myeloid (myeloperoxidase [MPO]) but not lymphoid-affiliated (CD3, Thy-1, and lambda5) genes. Additionally, most cells coexpress the stem cell-associated transcriptional regulators AML-1, PU.1, GATA-2 and Lmo2, as well as the granulocyte colony-stimulating factor receptor (G-CSF-R). These results show that the CD34(+)c-kit(+) population from the AGM represents a highly enriched source of multipotent hematopoietic cells, and suggest that limited coactivation of distinct lineage-affiliated genes is an early event in the generation of hematopoietic stem and progenitor cells during ontogeny. PMID- 10477674 TI - Activation of granulocyte-macrophage colony-stimulating factor and interleukin-3 receptor subunits in a multipotential hematopoietic progenitor cell line leads to differential effects on development. AB - Activation of specific cytokine receptors promotes survival and proliferation of hematopoietic progenitor cells but their role in the control of differentiation is unclear. To address this issue, the effects of human interleukin-3 (hIL-3) and human granulocyte-macrophage colony-stimulating factor (hGM-CSF) on hematopoietic development were investigated in hematopoietic progenitor cells. Murine multipotent factor-dependent cell-Paterson (FDCP)-mix cells, which can self-renew or differentiate, were transfected with the genes encoding the unique alpha and/or shared beta(c) human hIL-3 receptor (hIL-3 R) or hGM-CSF receptor (hGM R) subunits by retroviral gene transfer. Selective activation of hIL-3 Ralpha,beta(c) or hGM Ralpha,beta(c) transfects by hIL-3 and hGM-CSF promoted self-renewal and myeloid differentiation, respectively, over a range of cytokine (0.1 to 100 ng/mL) concentrations. These qualitatively distinct developmental outcomes were associated with different patterns of protein tyrosine phosphorylation and, thus, differential signaling pathway activation. The cell lines generated provide a model to investigate molecular events underlying self renewal and differentiation and indicate that the alpha subunits act in combination with the hbeta(c) to govern developmental decisions. The role of the alpha subunit in conferring specificity was studied by using a chimeric receptor composed of the extracellular hIL-3 Ralpha and intracellular hGM Ralpha subunit domains. This receptor promoted differentiation in response to hIL-3. Thus, the alpha subunit cytosolic domain is an essential component in determining cell fate via specific signaling events. PMID- 10477675 TI - The evidence-based analysis of treatment for chronic myeloid leukemia: An introduction to its methods and clinical implications. PMID- 10477676 TI - An evidence-based analysis of the effect of busulfan, hydroxyurea, interferon, and allogeneic bone marrow transplantation in treating the chronic phase of chronic myeloid leukemia: developed for the American Society of Hematology. AB - Because there are differing opinions regarding treatment of patients in the chronic phase of chronic myeloid leukemia (CML), the American Society of Hematology convened an expert panel to review and document evidence-based benefits and harms of treatment of CML with busulfan (BUS), hydroxyurea (HU), recombinant interferon-alpha (rIFN-alpha), and bone marrow transplantation (BMT). The primary measure for defining efficacy was survival. Analysis indicated a survival advantage for HU over BUS. Observational studies of rIFN-alpha suffer from numerous biases including sample size, variations in study populations, definitions of hematologic and cytogenetic remissions, and dose. That rIFN-alpha is more efficacious than chemotherapy is demonstrated by 6 prospective randomized trials. For patients with favorable clinical features in chronic phase, compared to HU and BUS, rIFN-alpha improves survival by a median of about 20 months. Most evidence suggests that rIFN-alpha is most effective when combined with other drugs and when given during the earliest stage of the chronic phase. Adding cytarabine to rIFN-alpha adds further survival benefit but increases toxicity. Limitations for evaluating the long-term benefits of allogeneic BMT include the retrospective nature of most studies, incomplete documentation of the clinical characteristics of the patients, paucity of the details on patient selection, lack of control groups, and limitations of survival calculations. Survival curves for BMT show that at least half of the patients transplanted remain alive 5 to 10 years after treatment, whereas similar curves for rIFN-alpha show a continuous relapse rate over time with the curves crossing at about 7 to 8 years. Estimates of long-term survival may be confounded by the selection biases mentioned and the analytic methods used. The magnitude of the incremental increase in benefit with BMT must be weighed against the potential serious harm and death that may accompany the procedure in the short term. The best results with BMT have been obtained when it is performed within 1 to 2 years from diagnosis. Since each treatment option involves tradeoffs between benefit and harm, patient choice must be based on the examination of facts presented in an unbiased fashion. Newly diagnosed younger patients and older patients who are candidates for BMT should also be offered information about IFN-based regimens, the tradeoffs involved, and, if possible, share in the treatment decision. Hopefully this analysis will provide the stimulus for evaluation of other important aspects of CML. PMID- 10477677 TI - Association of chromosome arm 9p abnormalities with adverse risk in childhood acute lymphoblastic leukemia: A report from the Children's Cancer Group. AB - Cytogenetic abnormalities of chromosome arm 9p occur frequently in children with acute lymphoblastic leukemia (ALL). We analyzed 201 such cases (11%) in 1,839 children with newly diagnosed ALL treated between 1989 and 1995 on risk-adjusted protocols of the Children's Cancer Group (CCG). The majority of patients (131; 65%) with a 9p abnormality were classified as higher risk. Nearly all patients had complex karyotypes; most cases had deletions of 9p, add/der(9p), a dicentric involving chromosome arm 9p, and/or balanced translocations and inversions involving 9p. Event-free survival (EFS) estimates at 6 years for patients with and without a 9p aberration were 61% (standard deviation [SD] = 5%) and 76% (SD = 2%; P <.0001). In addition, patients with a 9p abnormality had an increased cumulative incidence of both marrow (P =.04) and central nervous system (P =.0001) relapses. Overall survival also was significantly worse for patients with an abnormal 9p (P <.0001). These effects were most pronounced in standard-risk patients (age 1 to 9 years with white blood cell count <50,000/microL): 6-year EFS of 61% (SD = 9%) versus 80% (SD = 2%; P <.0001). Also, a 9p aberration was an adverse risk factor for B-lineage, but not T-lineage patients. The effect of 9p status on EFS was attenuated, but maintained in a multivariate analysis of EFS after adjustment for Philadelphia chromosome status, age, white blood cell (WBC) count, sex, race, and ploidy group (P =.01). Thus, abnormalities of chromosome arm 9p identify a subgroup of standard-risk patients with increased risk of treatment failure. PMID- 10477678 TI - Inheritance of chromosomally integrated human herpesvirus 6 DNA. AB - Human herpesvirus 6 (HHV-6) genome has been detected in several human lymphoproliferative disorders with no signs of active viral infection, and found to be integrated into chromosomes in some cases. We previously reported a woman with HHV-6-infected Burkitt's lymphoma. Fluorescence in situ hybridization showed that the viral genome was integrated into the long arm of chromosome 22 (22q13). The patient's asymptomatic husband also carried HHV-6 DNA integrated at chromosome locus 1q44. To assess the possibility of chromosomal transmission of HHV-6 DNA, we looked for HHV-6 DNA in the peripheral blood of their daughter. She had HHV-6 DNA on both chromosomes 22q13 and 1q44, identical to the site of viral integration of her mother and father, respectively. The findings suggested that her viral genomes were inherited chromosomally from both parents. The 3 family members were all seropositive for HHV-6, but showed no serological signs of active infection. To confirm the presence of HHV-6 DNA sequences, we performed polymerase chain reaction (PCR) with 7 distinct primer pairs that target different regions of HHV-6. The viral sequences were consistently detected by single-step PCR in all 3 family members. We propose a novel latent form for HHV 6, in which integrated viral genome can be chromosomally transmitted. The possible role of the chromosomally integrated HHV-6 in the pathogenesis of lymphoproliferative diseases remains to be explained. PMID- 10477679 TI - Safety of hydroxyurea in children with sickle cell anemia: results of the HUG KIDS study, a phase I/II trial. Pediatric Hydroxyurea Group. AB - Previous studies have determined the short-term toxicity profile, laboratory changes, and clinical efficacy associated with hydroxyurea (HU) therapy in adults with sickle cell anemia. The safety and efficacy of this agent in pediatric patients with sickle cell anemia has not been determined. Children with sickle cell anemia, age 5 to 15 years, were eligible for this multicenter Phase I/II trial. HU was started at 15 mg/kg/d and escalated to 30 mg/kg/d unless the patient experienced laboratory toxicity. Patients were monitored by 2-week visits to assess compliance, toxicity, clinical adverse events, growth parameters, and laboratory efficacy associated with HU treatment. Eighty-four children were enrolled between December 1994 and March 1996. Sixty-eight children reached maximum tolerated dose (MTD) and 52 were treated at MTD for 1 year. Significant hematologic changes included increases in hemoglobin concentration, mean corpuscular volume, mean corpuscular hemoglobin, and fetal hemoglobin parameters, and decreases in white blood cell, neutrophil, platelet, and reticulocyte counts. Laboratory toxicities typically were mild, transient, and were reversible upon temporary discontinuation of HU. No life-threatening clinical adverse events occurred and no child experienced growth failure. This Phase I/II trial shows that HU therapy is safe for children with sickle cell anemia when treatment was directed by a pediatric hematologist. HU in children induces similar laboratory changes as in adults. Phase III trials to determine if HU can prevent chronic organ damage in children with sickle cell anemia are warranted. PMID- 10477680 TI - Sickle cell acute chest syndrome: pathogenesis and rationale for treatment. AB - Acute chest syndrome (ACS) is a leading cause of death in sickle cell disease (SCD). Our previous work showed that hypoxia enhances the ability of sickle erythrocytes to adhere to human microvessel endothelium via interaction between very late activation antigen-4 (VLA4) expressed on sickle erythrocytes and the endothelial adhesion molecule vascular cell adhesion molecule-1 (VCAM-1). Additionally, hypoxia has been shown to decrease the production of nitric oxide (NO) which inhibits VCAM-1 upregulation. Based on these observations, we hypothesize that during ACS, the rapidly progressive clinical course that can occur is caused by initial hypoxia-induced pulmonary endothelial VCAM-1 upregulation that is not counterbalanced by production of cytoprotective mediators, including NO, resulting in intrapulmonary adhesion. We assessed plasma NO metabolites and soluble VCAM-1 in 36 patients with SCD and 23 age-matched controls. Patients with SCD were evaluated at baseline (n = 36), in vaso occlusive crisis (VOC; n = 12), and during ACS (n = 7). We observed marked upregulation of VCAM-1 during ACS (1,290 +/- 451 ng per mL; mean +/- 1 SD) with values significantly higher than controls (P <.0001) or patients either in steady state or VOC (P <. 01). NO metabolites were concomitantly decreased during ACS (9.2 +/- 1.5 nmol/mL) with values lower than controls (22.2 +/- 5.5), patients during steady state (21.4 +/- 5.5), or VOC (14.2 +/- 1.2) (P <.0001). Additionally, the ratio of soluble VCAM-1 to NO metabolites during ACS (132.9 +/- 46.5) was significantly higher when compared with controls (P <.0001) or patients either in steady state or VOC (P <.0001). Although hypoxia enhanced in vitro sickle erythrocyte-pulmonary microvessel adhesion, NO donors inhibited this process with concomitant inhibition of VCAM-1. We suggest that in ACS there is pathologic over expression of endothelial VCAM-1. Our investigations also provide a rationale for the therapeutic use in ACS of cytoprotective modulators including NO and dexamethasone, which potentially exert their efficacy by an inhibitory effect on VCAM-1 and concomitant inhibition of sickle erythrocyte-endothelial adhesion. PMID- 10477681 TI - Interleukin-1beta and tumor necrosis factor-alpha stimulate DNA binding of hypoxia-inducible factor-1. AB - The rate of transcription of several genes encoding proteins involved in O(2) and energy homeostasis is controlled by hypoxia-inducible factor-1 (HIF-1), a heterodimeric DNA binding complex composed of alpha and beta subunits. HIF-1 is considered the primary trans-acting factor for the erythropoietin (EPO) and vascular endothelial growth factor (VEGF) genes. Since EPO gene expression is inhibited by the proinflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), while no such effect has been reported with respect to the VEGF gene, we investigated the effects of IL-1beta and TNF-alpha on the activation of the HIF-1 DNA-binding complex and the amount of HIF-1alpha protein in human hepatoma cells in culture. Under normoxic conditions, both cytokines caused a moderate activation of HIF-1 DNA binding. In hypoxia, cytokines strongly increased HIF-1 activity compared with the effect of hypoxia alone. Only IL-1beta increased HIF-1alpha protein levels. In transient transfection experiments, HIF-1-driven reporter gene expression was augmented by cytokines only under hypoxic conditions. In contrast to their effect on EPO synthesis, neither IL-1beta nor TNF-alpha decreased VEGF production. The mRNA levels of HIF-1alpha and VEGF were unaffected. Thus, cytokine-induced inhibition of EPO production is not mediated by impairment of HIF-1 function. We propose that HIF-1 may be involved in modulating gene expression during inflammation. PMID- 10477682 TI - Phosphatidylinositol 3-kinase is involved in the protection of primary cultured human erythroid precursor cells from apoptosis. AB - Little is known about the physiologic role of phosphatidylinositol 3-kinase (PI 3K) in the development of erythrocytes. Previous studies have shown that the effects of the PI-3K inhibitor wortmannin on erythropoietin (EPO)-dependent cell lines differed depending on the cell type used. Wortmannin inhibited EPO-induced differentiation of some cell lines without affecting their proliferation; however, the EPO-induced proliferation of other cell lines was inhibited by wortmannin. In neither case were signs of apoptosis observed. We have previously reported that signaling in highly purified human colony forming units-erythroid (CFU-E), generated in vitro from CD34(+) cells, differed from that in EPO dependent cell lines. In the current study, we examined the effects of a more specific PI-3K inhibitor (LY294002) on human CFU-E. We found that LY294002 dose dependently inhibits the proliferation of erythroid progenitor cells with a half maximal effect at 10 micromol/L LY294002. LY294002 at similar concentrations also induces apoptosis of these cells, as evidenced by the appearance of annexin V binding cells and DNA fragmentation. The steady-state phosphorylation of AKT at Ser-473 that occurs as a result of PI-3K activation was also inhibited by LY294002 at similar concentrations, suggesting that the effects of LY294002 are specific. Interestingly, the acceleration of apoptosis by LY294002 was observed in the presence or absence of EPO. Further, deprivation of EPO resulted in accelerated apoptosis irrespective of the presence of LY294002. Our study confirms and extends the finding that signaling in human primary cultured erythroid cells is significantly different from that in EPO-dependent cell lines. These data suggest that PI-3K has an antiapoptotic role in erythroid progenitor cells. In addition, 2 different pathways for the protection of primary erythroid cells from apoptosis likely exist: 1 independent of EPO that is LY294002 sensitive and one that is EPO-dependent and at least partly insensitive to LY294002. PMID- 10477683 TI - c-Maf induces monocytic differentiation and apoptosis in bipotent myeloid progenitors. AB - The transcriptional mechanisms that drive colony-forming unit granulocyte macrophage (CFU-GM) myeloid progenitors to differentiate into cells of either the granulocytic or monocytic lineage are not fully understood. We have shown that the c-Maf and c-Myb transcription factors physically interact in myeloid cells to form inhibitory complexes that hinder transactivation of c-Myb target genes through direct binding to Myb consensus sites. These complexes arise in a developmentally regulated pattern, peaking at the promyelocyte stage, or in cell model systems, appearing soon after the induction of monocytic differentiation. We wished to determine if this developmentally related interaction is a consequence of myeloid differentiation or an intrinsic differentiating stimulus. Because the elevated Myb:Maf status seen in differentiating cells can be recapitulated by overexpression of c-Maf in myeloid cell lines, we inducibly expressed the c-Maf cDNA in 2 bipotent human myeloid progenitor cells. Elevated levels of c-Maf protein led to marked increases in Myb:Maf complexes and the accumulation of monocyte/macrophage cells, followed by eventual programmed cell death. Analysis of targets that could mediate these phenotypic changes indicated that c-Maf likely plays a key role in myeloid cell development through dual mechanisms; inhibition of a select set of c-Myb regulated targets, such as Bcl-2 and CD13/APN, coupled with the activation of as yet undefined differentiation promoting genes. PMID- 10477684 TI - Lineage-specific expression of human immunodeficiency virus (HIV) receptor/coreceptors in differentiating hematopoietic precursors: correlation with susceptibility to T- and M-tropic HIV and chemokine-mediated HIV resistance. AB - Human immunodeficiency virus (HIV) entry is mediated not only by the CD4 receptor, but also by interaction with closely related molecules that act as membrane coreceptors. We have analyzed mRNA expression and/or cell membrane exposition of the coreceptors most widely used by diverse HIV-1 strains (CXCR4, CCR5, and CCR3) on purified hematopoietic progenitor cells (HPCs) induced in liquid suspension culture to unilineage differentiation/maturation through the erythroid (E), granulocytic (G), megakaryocytic (Mk), and monocytic (Mo) lineages. Reverse transcriptase-polymerase chain reaction (RT-PCR) and cytofluorimetric analysis showed the presence of both CXCR4 and CCR5 in quiescent HPCs, but failed to detect CCR3-specific transcripts. Chemokine expression in HPC progenies showed that CXCR4 receptor is detected on the majority of MKs from early to late stages of maturation, whereas it is moderately decreased in the Mo lineage. In the G pathway, two distinct cell populations, CXCR4(+) and CXCR4(-), were observed: morphological analysis of the sorted populations showed that the CXCR4(+) cells were largely eosinophils and the CXCR4(-) were granulocytes of the neutrophilic series. Furthermore, in the E pathway, CXCR4 was almost completely absent. CCR5 expression is restricted to Mo cultures, ie, approximately 30% to 80% cells throughout all monocytopoietic differentiation/maturation stages. Finally, CCR3 mRNA is always absent in all the unilineage cultures. Evaluation of CD4 expression by flow cytometry on both quiescent HPCs and differentiating unilineage precursors showed that the CD4 receptor is present on approximately 15% of the starting CD34(+) HPC population, highly expressed in the Mo lineage up to 80% at terminal maturation, present on 20% to 30% of maturing Mks, and not detectable in either the E or G lineage. Expression of CD4 receptor together with CXCR4 and/or CCR5 coreceptor in the four lineages correlates with hematopoietic precursor susceptibility to T-lymphotropic and macrophage (M)-tropic HIV strains infection: (1) CD4(-) G and E cells were resistant to both M-tropic and T lymphotropic strains; (2) HPC-derived Mks were susceptible to T-tropic, but resistant to M-tropic, infection; (3) Mo differentiating cells efficiently replicate both HIV strains. Furthermore, we showed that the CXCR4 and CCR5 ligands (stromal-derived factor 1 and macrophage-inflammatory protein-1alpha [MIP 1alpha], MIP-1beta and RANTES, respectively) inhibit HIV replication in both maturing Mo and Mk cells. Taken together, our data show a lineage-specific modulation of chemokine receptor/coreceptor during hematopoietic cell differentiation and extend previous observations on the relationship between the expression of HIV receptor/coreceptors, susceptibility, and chemokine-mediated resistance to HIV infection. PMID- 10477685 TI - Megakaryocyte growth and development factor-induced proliferation and differentiation are regulated by the mitogen-activated protein kinase pathway in primitive cord blood hematopoietic progenitors. AB - In several erythroleukemia cell lines, activation of mitogen-activated protein kinases (MAPK) by phorbol esters or megakaryocyte growth and development factor (MGDF) is required for induction of megakaryocytic phenotype and growth arrest. To support this model, we have examined the effect of a specific inhibitor of this pathway (PD98059) on human CD34(+) hematopoietic progenitors isolated from cord blood (CB), induced to differentiate along the megakaryocytic lineage in liquid cultures supplemented with rhuMGDF. RhuMGDF induced a sustained activation of MAPK in megakaryocytes and this activation was completely inhibited in the presence of low concentrations of PD98059 (6 to 10 micromol/L). At this concentration, PD98059 induced an increase in cell proliferation, resulting in accumulation of viable cells and a prolongation of the life time of the cultures. This increase correlated with an increase in DNA synthesis rather than with a reduction in apoptosis. This effect was combined with developmental changes indicative of delayed megakaryocytic differentiation: (1) PD98059-treated cells tended to retain markers of immature progenitors as shown by the increased proportion of both CD34(+) and CD41(+)CD34(+) cells. (2) PD98059-treated cultures were greatly enriched in immature blasts cells. (3) PD98059 increased megakaryocytic progenitors able to form colonies in semisolid assays. Thus, the MAPK pathway, although not required for megakaryocyte formation, seems to be involved in the transition from proliferation to maturation in megakaryocytes. Inhibition of MAPK activation also led to an increase in the number and size of erythroid colonies without affecting granulocyte/macrophage progenitor numbers suggesting that, in addition to the megakaryocytic lineage, the MAPK pathway could play a role in erythroid lineage differentiation. PMID- 10477686 TI - Heterodimerization of the alpha and beta chains of the interleukin-3 (IL-3) receptor is necessary and sufficient for IL-3-induced mitogenesis. AB - The high-affinity receptor for interleukin-3 (IL-3) is a complex of the IL-3 binding subunit (alpha(IL-3)) and a larger beta chain-beta(c), or, in the mouse, beta(c) or its close relative beta(IL-3). There is evidence that the critical event that initiates signaling is not the approximation of the cytoplasmic domains of alpha(IL-3) and beta(IL-3), but is, rather, the formation of a beta beta homodimer. Many of these studies involved the analyses of receptor chimeras where the cytoplasmic domains were derived from alpha(IL-3), beta(c) or beta(IL 3), and the extracellular domains were derived from other cytokine receptors, such as the erythropoietin receptor (EpoR). However, evidence that the EpoR may also associate with other receptors clouds the interpretation of these experiments. Therefore, we reevaluated the structure of the functional IL-3R using chimeric receptors with extracellular domains derived not from members of the cytokine-receptor family, but from CD8 or CD16. We show, by expression of these chimeras in Ba/F3 or CTLL-2 cells, that mitogenic signals were only generated by heterodimerization of the cytoplasmic domains of alpha(IL-3) and beta(IL-3). Homodimers of either alpha(IL-3) or beta(IL-3), alone or in combination, were nonfunctional. Furthermore, the ability of heterodimers to stimulate mitogenesis correlated with their ability to induce tyrosine phosphorylation of JAK-2. These data suggest that the physiological activation of the IL-3R involves the generation of simple heterodimers of alpha(IL-3) and beta(IL-3). PMID- 10477687 TI - Long-term ex vivo maintenance and expansion of transplantable human hematopoietic stem cells. AB - We have developed a stromal-based in vitro culture system that facilitates ex vivo expansion of transplantable CD34(+) thy-1(+) cells using long-term hematopoietic reconstitution in severe combined immunodeficient-human (SCID-hu) mice as an in vivo assay for transplantable human hematopoietic stem cells (HSCs). The addition of leukemia inhibitory factor (LIF) to purified CD34(+) thy 1(+) cells on AC6.21 stroma, a murine bone marrow-derived stromal cell line, caused expansion of cells with CD34(+) thy-1(+) phenotype. Addition of other cytokines, including interleukin-3 (IL-3), IL-6, granulocyte-macrophage colony stimulating factor, and stem cell factor, to LIF in the cultures caused a 150 fold expansion of cells retaining the CD34(+) thy-1(+) phenotype. The ex vivo expanded CD34(+) thy-1(+) cells gave rise to multilineage differentiation, including myeloid, T, and B cells, when transplanted into SCID-hu mice. Both murine LIF (cannot bind to human LIF receptor) and human LIF caused expansion of human CD34(+) thy-1(+) cells in vitro, suggesting action through the murine stroma. Furthermore, another human HSC candidate, CD34(+) CD38(-) cells, shows a similar pattern of proliferative response. This suggests that ex vivo expansion of transplantable human stem cells under this in vitro culture system is a general phenomenon and not just specific for CD34(+) thy-1(+) cells. PMID- 10477688 TI - Evidence for extracellular processing of pro-von Willebrand factor after infusion in animals with and without severe von Willebrand disease. AB - Although proteolytic processing of pro-von Willebrand factor (pro-vWF) resulting in free propeptide and mature vWF is known to be initiated intracellularly, vWF released from endothelial cells may contain a high proportion of incompletely processed pro-vWF. Because pro-vWF is only rarely detectable in normal human plasma, we investigated whether extracellular processing of pro-vWF is possible. A recombinant preparation (rpvWF) containing both pro-vWF and mature vWF subunits was infused into 2 pigs and 1 dog with severe von Willebrand disease, 2 mice with a targeted disruption of the vWF gene, and 2 healthy baboons. Total vWF antigen (vWF:Ag), free propeptide, and pro-vWF were measured using enzyme-linked immunosorbent assay techniques in blood samples drawn before and after infusion. vWF:Ag increased promptly. No pro-vWF could be detected when the first postinfusion sample was drawn after 30 minutes (pigs) or 60 minutes (mice), but pro-vWF was detectable for short periods when postinfusion samples were drawn after 15 minutes (dog) or 5 minutes (baboons). In contrast, free propeptide was increased at the first timepoint measured, suggesting that it was generated from the pro-vWF in the rpvWF preparation. vWF multimers were analyzed in the rpvWF preparation and in plasma samples drawn before and after infusion of rpvWF using ultra-high resolution 3% agarose gels to allow separation of homo- and hetero forms of the vWF polymers. Within 30 minutes after infusion in the pigs, 1 hour in the dog and the mice, and within 2 hours in the baboons, the multimer pattern had changed to that typically seen in mature vWF. These data indicate that propeptide cleavage from unprocessed vWF can occur extracellularly in the circulation. The enzyme or enzymes responsible for this cleavage in plasma remain to be identified. PMID- 10477689 TI - Glycoprotein Ib-V-IX, a receptor for von Willebrand factor, couples physically and functionally to the Fc receptor gamma-chain, Fyn, and Lyn to activate human platelets. AB - The adhesion molecule von Willebrand factor (vWF) activates platelets upon binding 2 surface receptors, glycoprotein (GP) Ib-V-IX and integrin alpha(IIb)beta(3). We have used 2 approaches to selectively activate GP Ib using either the snake venom lectin alboaggregin-A or mutant recombinant forms of vWF (triangle upA1-vWF and RGGS-vWF) with selective binding properties to its 2 receptors. We show that activation of GP Ib induces platelet aggregation, secretion of 5-hydroxy tryptamine (5-HT), and an increase in cytosolic calcium. Syk becomes tyrosine phosphorylated and activated downstream of GP Ib, and associates with several tyrosine-phosphorylated proteins including the Fc receptor gamma-chain through interaction with Syk SH2 domains. GP Ib physically associates with the gamma-chain in GST-Syk-SH2 precipitates from platelets stimulated through GP Ib, and 2 Src family kinases, Lyn and Fyn, also associate with this signaling complex. In addition, GP Ib stimulation couples to tyrosine phosphorylation of phospholipase Cgamma2. The Src family-specific inhibitor PP1 dose-dependently inhibits phosphorylation of Syk, its association with tyrosine phosphorylated gamma-chain, phosphorylation of PLCgamma2, platelet aggregation, and 5-HT release. The results indicate that, upon activation, GP Ib is physically associated with FcR gamma-chain and members of the Src family kinases, leading to phosphorylation of the gamma-chain, recruitment, and activation of Syk. Phosphorylation of PLCgamma2 also lies downstream of Src kinase activation and may critically couple early signaling events to functional platelet responses. PMID- 10477690 TI - Discordant expression of tissue factor and its activity in polarized epithelial cells. Asymmetry in anionic phospholipid availability as a possible explanation. AB - Recent studies have shown a discrepancy between the level of tissue factor (TF) expression and the level of TF procoagulant activity on the apical and basolateral surface domains of polarized epithelial cells. The present investigation was performed to elucidate possible reasons for the discordant expression of TF and its activity on the surface of polarized epithelial cells using a human intestinal epithelial cell line, Caco-2 and Madin-Darby canine kidney epithelial cells, type II (MDCK-II). Functional activity of coagulation factor VIIa (VIIa) in complex with TF was 6- to 7-fold higher on the apical than the basolateral surface in polarized Caco-2 cells. In contrast, no significant difference was found in the formation of TF/VIIa complexes between the apical and basolateral surface. Confocal microscopy of Caco-2 cells showed TF expression on both the apical and the basolateral surface domains. Studies with MDCK-II cells showed that the specific functional activity of TF expressed on the apical cell surface was 5-fold higher than on the basolateral surface. To test whether differential expression of TF pathway inhibitor (TFPI) on the apical and basolateral surface could account for differences in TF/VIIa functional activity, we measured cell-surface-bound TFPI activity in Caco-2 cells. Small but similar amounts of TFPI were found on both surfaces. Further, addition of inhibitory anti TFPI antibodies induced a similar enhancement of TF/VIIa activity on both surface domains. Because the availability of anionic phospholipids on the outer leaflet of the cell membrane could regulate TF/VIIa functional activity, we measured the distribution of anionic phospholipids on the apical and basolateral surface by annexin V binding and thrombin generation. The results showed that the anionic phospholipid content on the basolateral surface, compared with the apical surface, was 3- to 4-fold lower. Mild acid treatment of polarized Caco-2 cells, which markedly increased the anionic phospholipid content on the basolateral surface membrane, increased the TF/VIIa activity on the basolateral surface without affecting the number of TF/VIIa complexes formed on the surface. Overall, our data suggest that an uneven expression of TF/VIIa activity between the apical and basolateral surface of polarized epithelial cells is caused by differences in anionic phospholipid content between the two surface domains and not from a polar distribution of TFPI. PMID- 10477691 TI - Dichotomous regulation of myosin phosphorylation and shape change by Rho-kinase and calcium in intact human platelets. AB - Both Rho-kinase and the Ca(2+)/calmodulin-dependent myosin light chain (MLC) kinase increase the phosphorylation of MLC. We show that upon thrombin receptor stimulation by low-dose thrombin or the peptide ligand YFLLRNP, or upon thromboxane receptor activation by U46619, shape change and MLC phosphorylation in human platelets proceed through a pathway that does not involve an increase in cytosolic Ca(2+). Under these conditions, Y-27632, a specific Rho-kinase inhibitor, prevented shape change and reduced the stimulation of MLC phosphorylation. In contrast, Y-27632 barely affected shape change and MLC phosphorylation by adenosine diphosphate (ADP), collagen-related peptide, and ionomycin that were associated with an increase in cytosolic Ca(2+) and inhibited by BAPTA-AM/EGTA treatment. Furthermore, C3 exoenzyme, which inactivates Rho, inhibited preferentially the shape change induced by YFLLRNP compared with ADP and ionomycin. The results indicate that the Rho/Rho-kinase pathway is pivotal in mediating the MLC phosphorylation and platelet shape change by low concentrations of certain G protein-coupled platelet receptors, independent of an increase in cytosolic Ca(2+). Our study defines 2 alternate pathways, Rho/Rho-kinase and Ca(2+)/calmodulin-regulated MLC-kinase, that lead independently of each other through stimulation of MLC-phosphorylation to the same physiological response in human platelets (ie, shape change). PMID- 10477692 TI - Induction of decay-accelerating factor by cytokines or the membrane-attack complex protects vascular endothelial cells against complement deposition. AB - Vascular endothelium is continuously exposed to complement-mediated challenge, and this is enhanced during inflammation. Although the complement-regulatory proteins decay-accelerating factor (DAF), CD59, and membrane cofactor protein (MCP) protect endothelial cells (ECs) against complement-mediated injury, the control of their expression and relative contributions to vascular protection is unclear. We explored the hypothesis that mechanisms exist which induce upregulation of complement-regulatory proteins on ECs to maintain vascular function in inflammation. Tumor necrosis factor alpha (TNFalpha) and interferon gamma (IFNgamma) each increased DAF expression but not CD59 or MCP expression, and a combination of these cytokines was more potent than either alone. Cytokine induced expression depended on increased DAF mRNA and de novo protein synthesis and was maximal by 72 hours. In addition, assembly of the membrane-attack complex (MAC) on ECs induced a 3-fold increase in DAF expression, and this was enhanced by cytokines. DAF upregulation was not inhibited by protein kinase C (PKC) antagonists. The increase in DAF was functionally relevant since it reduced complement 3 (C3) deposition by 40%, and this was inhibited by an anti-DAF monoclonal antibody. These observations indicate that upregulation of DAF expression by cytokines or MAC may represent an important feedback mechanism to maintain the integrity of the microvasculature during subacute and chronic inflammatory processes involving complement activation. PMID- 10477693 TI - Calpain functions in a caspase-independent manner to promote apoptosis-like events during platelet activation. AB - Apoptosis and platelet activation share common morphological and biochemical features. Because caspases are essential mediators of apoptosis, we examined whether platelets contain these proteinases and use them during platelet activation. Human platelets contained caspase-9, caspase-3, and the caspase activators APAF-1 and cytochrome c as shown by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blotting. Upon treatment with cytochrome c and dATP, platelet cytoplasmic extracts recapitulated apoptotic events, including sequential activation of procaspase-9 and procaspase-3 and subsequent proteolysis of caspase substrates. Calcium ionophore-stimulated platelets also recapitulated apoptotic events, including cell shrinkage, plasma membrane microvesiculation, phosphatidyl serine externalization, and proteolysis of procaspase-9, procaspase-3, gelsolin, and protein kinase C-delta. Strikingly, however, these events occurred without caspase activation or release of mitochondrial cytochrome c, suggesting a role for a noncaspase proteinase. Supporting this, inhibition of the calcium-dependent proteinase, calpain, prevented caspase proteolysis, 'apoptotic' substrate cleavage, and platelet microvesiculation. In vitro, purified calpain cleaved recombinant procaspase-9 and procaspase-3 without activating either caspase, confirming the inhibitor studies. These data implicate calpain as a potential regulator of caspases and suggest that calpain, not caspases, promotes apoptosis-like events during platelet activation. PMID- 10477694 TI - Unique antiplatelet effects of a novel S-nitrosoderivative of a recombinant fragment of von Willebrand factor, AR545C: in vitro and ex vivo inhibition of platelet function. AB - The recombinant fragment of von Willebrand factor (vWF) spanning Ala444 to Asp730 and containing an Arg545Cys mutation (denoted AR545C) has antithrombotic properties that are principally a consequence of its ability to inhibit platelet adhesion to subendothelial matrix. Endothelial-derived nitric oxide (NO) can also inhibit platelet function, both as a consequence of inhibiting adhesion as well as activation and aggregation. Nitric oxide can react with thiol functional groups in the presence of oxygen to form S-nitrosothiols, which are naturally occurring NO derivatives that prolong the biological actions of NO. Because AR545C has a single free cysteine (Cys545), we attempted to synthesize the S nitroso-derivative of AR545C and to characterize its antiplatelet effects. We successfully synthesized S-nitroso-AR545C and found that it contained 0.96 mol S NO per mole peptide. S-nitroso-AR545C was approximately 5-fold more potent at inhibiting platelet agglutination than was the unmodified peptide (IC(50) = 0.02 +/- 0. 006 micromol/L v 0.1 +/- 0.03 micromol/L, P =.001). In addition and by contrast, S-nitroso-AR545C was a powerful inhibitor of adenosine diphosphate induced platelet aggregation (IC(50) = 0.018 +/- 0.002 micromol/L), while AR545C had no effect on aggregation. These effects were confirmed in studies of adhesion to and aggregation on extracellular matrix under conditions of shear stress in a cone-plate viscometer, where 1.5 micromol/L S-nitroso-AR545C inhibited platelet adhesion by 83% and essentially completely inhibited aggregate formation, while the same concentration of AR545C inhibited platelet adhesion by 74% and had significantly lesser effect on aggregate formation on matrix (P /=730 days) and were tested for susceptibility to spontaneous apoptosis and anti Fas triggered apoptosis in vitro. Substantial proportions of CD4(+) and CD8(+) cells generated during the first year after transplantation, but not by day 730, exhibited in these assays decreased mitochondrial membrane potential (triangle upPsim) and apoptotic DNA fragmentation. The apoptotic phenotype tended to disappear late in the follow-up period, when substantial absolute numbers of naive (CD45RA(+)/CD62-L(+)) T cells had repopulated the peripheral blood compartment of the BMT patients. The rate of spontaneous cell death in vitro was significantly correlated with lower levels of ex vivo Bcl-2 protein, as assessed by cytofluorometry and Western blot analysis. In contrast, the levels of Bax protein remained unchanged, resulting in dysregulated Bcl-2/Bax ratios. Cell death primarily concerned the expanded CD8(+)/CD45R0(+) subpopulation, although CD45R0(-) subpopulations were also involved, albeit to a lesser extent. These results show that the T-cell regeneration/expansion occurring after BMT is accompanied by decreased levels of Bcl-2 and susceptibility to apoptosis. PMID- 10477708 TI - Detection of abnormal pretransplant clones in progenitor cells of patients who developed myelodysplasia after autologous transplantation. AB - Secondary myelodysplastic syndromes (MDS) have been reported after autologous transplantation. It is not known whether the MDS results from the pretransplant conventional-dose chemotherapy or from the high-dose chemotherapy (HDC) used for the transplant procedure. We performed a multicenter, retrospective analysis of morphologically normal pretransplant marrow or stem cell specimens from 12 patients who subsequently developed myelodysplasia after HDC. To determine if the abnormal clone was present before HDC, we used fluorescence in situ hybridization (FISH) to detect the cytogenetic markers observed at the onset of posttransplant MDS. Cryopreserved, pretransplant bone marrow, peripheral blood stem cell specimens, obtained at the time of harvest, or archival smears were used. Standard cytogenetic analysis had been performed pretransplant in four patients, showing a normal karyotype. In 9 of 12 cases, the same cytogenetic abnormality observed at the time of MDS diagnosis was detected by FISH in the pre-HDC specimens. Our findings support the hypothesis that, in many cases of posttransplant MDS, the stem cell damage results from prior conventional-dose chemotherapy and may be unrelated to HDC or the transplantation process itself. PMID- 10477709 TI - Fusion of a novel gene, BTL, to ETV6 in acute myeloid leukemias with a t(4;12)(q11-q12;p13). AB - The ETV6 gene (also known as TEL) is the main target of chromosomal translocations affecting chromosome band 12p13. The rearrangements fuse ETV6 to a wide variety of partner genes in both myeloid and lymphoid malignancies. We report here 4 new cases of acute myeloid leukemia (AML) with very immature myeloblasts (French-American-British [FAB]-M0) and with a t(4;12)(q11-q12;p13). In all cases, ETV6 was found recombined to a new gene, homologous to the mouse Brx gene. The gene was named BTL (Brx-like Translocated in Leukemia). Reverse transcriptase-polymerase chain reaction (RT-PCR) experiments indicate that the expression of the BTL-ETV6 transcript, but not of the reciprocal ETV6-BTL transcript, is a common finding in these leukemias. In contrast to the majority of other ETV6 fusions, both the complete helix-loop-helix (HLH) and ETS DNA binding domains of ETV6 are present in the predicted BTL-ETV6 fusion protein, and the chimeric gene is transcribed from the BTL promoter. PMID- 10477710 TI - Identification of the molecular genetic defect of patients with methemoglobin M Kankakee (M-Iwate), alpha87 (F8) His --> Tyr: evidence for an electrostatic model of alphaM hemoglobin assembly. AB - We determined that the molecular defect of 2 patients with hemoglobin (Hb) M Kankakee [Hb M-Iwate, alpha87 (F8) His --> Tyr] resides in the alpha1-globin gene. The proportion of Hb M observed is higher than that predicted for an alpha1 globin variant. Our evidence suggests that the greater-than-expected proportion of Hb M-Kankakee results from preferential association of the electronegative beta-globin chains with the alpha(M)-globin chains that are more electropositive than normal alpha-globin chains. PMID- 10477711 TI - Mutational status of Ig V(H) genes provides clinically valuable information in B cell chronic lymphocytic leukemia. PMID- 10477712 TI - Ig V gene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia. AB - Cellular immunophenotypic studies were performed on a cohort of randomly selected IgM(+) B-chronic lymphocytic leukemia (B-CLL) cases for which Ig V(H) and V(L) gene sequences were available. The cases were categorized based on V gene mutation status and CD38 expression and analyzed for treatment history and survival. The B-CLL cases could be divided into 2 groups. Those patients with unmutated V genes displayed higher percentages of CD38(+) B-CLL cells (>/=30%) than those with mutated V genes that had lower percentages of CD38(+) cells (<30%). Patients in both the unmutated and the >/=30% CD38(+) groups responded poorly to continuous multiregimen chemotherapy (including fludarabine) and had shorter survival. In contrast, the mutated and the <30% CD38(+) groups required minimal or no chemotherapy and had prolonged survival. These observations were true also for those patients who stratified to the Rai intermediate risk category. In the mutated and the <30% CD38(+) groups, males and females were virtually equally distributed, whereas in the unmutated and the >/=30% CD38(+) groups, a marked male predominance was found. Thus, Ig V gene mutation status and the percentages of CD38(+) B-CLL cells appear to be accurate predictors of clinical outcome in B-CLL patients. These parameters, especially CD38 expression that can be analyzed conveniently in most clinical laboratories, should be valuable adjuncts to the present staging systems for predicting the clinical course in individual B-CLL cases. Future evaluations of new therapeutic strategies and drugs should take into account the different natural histories of patients categorized in these manners. PMID- 10477713 TI - Unmutated Ig V(H) genes are associated with a more aggressive form of chronic lymphocytic leukemia. AB - Despite having several characteristics of naive B cells, chronic lymphocytic leukemia (CLL) cells have been shown in some cases to have somatically mutated Ig variable region genes, indicating that the cell of origin has passed through the germinal center. A previous study of patients with CLL found an association between lack of somatic mutation and trisomy 12 and, therefore, possibly with a less favorable prognosis. We have sequenced the Ig V(H) genes of the tumor cells of 84 patients with CLL and correlated our findings with clinical features. A total of 38 cases (45.2%) showed >/= 98% sequence homology with the nearest germline V(H) gene; 46 cases (54.8%) showed >2% somatic mutation. Unmutated V(H) genes were significantly associated with V1-69 and D3-3 usage, with atypical morphology; isolated trisomy 12, advanced stage and progressive disease. Survival was significantly worse for patients with unmutated V(H) genes irrespective of stage. Median survival for stage A patients with unmutated V(H) genes was 95 months compared with 293 months for patients whose tumors had mutated V(H) genes (P =.0008). The simplest explanation is that CLL comprises 2 different diseases with different clinical courses. One, arising from a memory B cell, has a benign course, the other, arising from a naive B cell, is more malignant. PMID- 10477715 TI - Regulation of the erythropoietin gene. PMID- 10477714 TI - Promoter elements of vav drive transgene expression in vivo throughout the hematopoietic compartment. AB - To develop a method for targeting expression of genes to the full hematopoietic system, we have used transgenic mice to explore the transcriptional regulation of the vav gene, which is expressed throughout this compartment but rarely outside it. Previously, we showed that a cluster of elements surrounding its promoter could drive hematopoietic-specific expression of a bacterial lacZ reporter gene, but the expression was confined to lymphocytes and was sporadically silenced. Those limitations are ascribed here to the prokaryotic reporter gene. With a human CD4 (hCD4) cell surface reporter, the vav promoter elements drove expression efficiently and stably in virtually all nucleated cells of adult hematopoietic tissues but not notably in nonhematopoietic cell types. In multiple lines, hCD4 appeared on most, if not all, B and T lymphocytes, granulocytes, monocytes, megakaryocytes, eosinophils, and nucleated erythroid cells. Moreover, high levels appeared on both lineage-committed progenitors and the more primitive preprogenitors. In the fetus, expression was evident in erythroid cells of the definitive but not the primitive type. These results indicate that a prokaryotic sequence can inactivate a transcription unit and that the vav promoter region constitutes a potent transgenic vector for the entire definitive hematopoietic compartment. PMID- 10477716 TI - Nuclear factor-kappaB-dependent induction of interleukin-8 gene expression by tumor necrosis factor alpha: evidence for an antioxidant sensitive activating pathway distinct from nuclear translocation. AB - Tumor necrosis factor alpha (TNFalpha) is a pluripotent activator of inflammation by inducing a proinflammatory cytokine cascade. This phenomenon is mediated, in part, through inducible expression of the CXC chemokine, interleukin-8 (IL-8). In this study, we investigate the role of TNFalpha-inducible reactive oxygen species (ROS) in IL-8 expression by "monocyte-like" U937 histiocytic lymphoma cells. TNFalpha is a rapid activator of IL-8 gene expression by U937, producing a 50 fold induction of mRNA within 1 hour of treatment. In gene transfection assays, the effect of TNFalpha requires the presence of an inducible nuclear factor kappaB (NF-kappaB) (Rel A) binding site in the IL-8 promoter. TNFalpha treatment induces a rapid translocation of the 65 kD transcriptional activator NF-kappaB subunit, Rel A, whose binding in the nucleus occurs before changes in intracellular ROS. Pretreatment (or up to 15 minutes posttreatment) relative to TNFalpha with the antioxidant dimethyl sulfoxide (DMSO) (2% [vol/vol]) blocks 80% of NF-kappaB-dependent transcription. Surprisingly, however, DMSO has no effect on inducible Rel A binding. Similar selective effects on NF-kappaB transcription are seen with the unrelated antioxidants, N-acetylcysteine (NAC) and vitamin C. These data indicate that TNFalpha induces a delayed ROS-dependent signalling pathway that is required for NF-kappaB transcriptional activation and is separable from that required for its nuclear translocation. Further definition of this pathway will yield new insights into inflammation initiated by TNFalpha signalling. PMID- 10477717 TI - Macrophage-derived chemokine is localized to thymic medullary epithelial cells and is a chemoattractant for CD3(+), CD4(+), CD8(low) thymocytes. AB - Macrophage-derived chemokine (MDC) is a recently identified CC chemokine that is a potent chemoattractant for dendritic cells, natural killer (NK) cells, and the Th2 subset of peripheral blood T cells. In normal tissues, MDC mRNA is expressed principally in the thymus. Immunohistochemical analysis performed on 5 human postnatal thymuses showed high MDC immunoreactivity, which was selectively localized to epithelial cells within the medulla. To examine the effects of MDC on immature T cells, we have identified cDNA clones for mouse and rat MDC. Expression of MDC in murine tissues is also highly restricted, with significant levels of mRNA found only in the thymus. Thymocytes express high-affinity binding sites for MDC (kd = 0.7 nmol/L), and, in vitro, MDC is a chemoattractant for these cells. MDC-responsive murine thymocytes express mRNA for CCR4, a recently identified receptor for MDC. Phenotypic analysis of MDC-responsive cells shows that they are enriched for a subset of double-positive cells that express high levels of CD3 and CD4 and that have reduced levels of CD8. This subset of MDC responsive cells is consistent with the observed expression of MDC within the medulla, because more mature cells are found there. MDC may therefore play a role in the migration of T-cell subsets during development within the thymus. PMID- 10477718 TI - CCR5 binds multiple CC-chemokines: MCP-3 acts as a natural antagonist. AB - CCR5 was first characterized as a receptor for MIP-1alpha, MIP-1beta, and RANTES, and was rapidly shown to be the main coreceptor for M-tropic human immunodeficiency virus (HIV)-1 strains and simian immunodeficiency virus (SIV). Chemokines constitute a rapidly growing family of proteins and receptor-chemokine interactions are known to be promiscuous and redundant. We have therefore tested whether other CC-chemokines could bind to and activate CCR5. All CC-chemokines currently available were tested for their ability to compete with [(125)I]-MIP 1beta binding on a stable cell line expressing recombinant CCR5, and/or to induce a functional response in these cells. We found that in addition to MIP-1beta, MIP 1alpha, and RANTES, five other CC-chemokines could compete for [(125)I]-MIP-1beta binding: MCP-2, MCP-3, MCP-4, MCP-1, and eotaxin binding was characterized by IC(50) values of 0.22, 2.14, 5.89, 29.9, and 21.7 nmol/L, respectively. Among these ligands, MCP-3 had the remarkable property of binding CCR5 with high affinity without eliciting a functional response, MCP-3 could also inhibit the activation of CCR5 by MIP-1beta and may therefore be considered as a natural antagonist for CCR5. It was unable to induce significant endocytosis of the receptor. Chemokines that could compete with high affinity for MIP-1beta binding could also compete for monomeric gp120 binding, although with variable potencies; maximal gp120 binding inhibition was 80% for MCP-2, but only 30% for MIP-1beta. MCP-3 could compete efficiently for gp120 binding but was, however, found to be a weak inhibitor of HIV infection, probably as a consequence of its inability to downregulate the receptor. PMID- 10477719 TI - Reconstitution of early lymphoid proliferation and immune function in Jak3 deficient mice by interleukin-3. AB - Expansion of early lymphoid progenitors requires interleukin-7 (IL-7), which functions through gamma(c)-mediated receptor activation of Jak3. Jak3 deficiency is a cause of severe combined immunodeficiency (SCID) in humans and mice. IL-3 activates many of the same signaling pathways as IL-7, such as Stat5, but achieves this effect through the activation of Jak2 rather than Jak3. We hypothesized that expansion of an IL-7-responsive precursor population through a Jak3-independent pathway using IL-3 may stimulate early lymphoid progenitors and restore lymphopoiesis in Jak3(-/-) mice. Newborn Jak3(-/-) mice that were injected with IL-3 demonstrated thymic enlargement, a 2- to 20-fold increase in thymocyte numbers, and up to a 10-fold expansion in the number of CD4(+), CD8(+), and B220(+)/IgM(+) splenic lymphocytes, consistent with an effect upon an early lymphoid progenitor population. In contrast to control mice, IL-3-treated Jak3(-/ ) mice challenged with the allogeneic major histocompatibility complex (MHC) class I-bearing tumor P815 developed a specific CD8-dependent cytotoxic T lymphocyte (CTL) response. IL-3-treated mice also mounted influenza-specific CTL responses and survival was prolonged. The beneficial effects of IL-3 are proposed to be produced by stimulation of a lymphoid precursor population of IL 7Ralpha(+)/IL-3Ralpha(+) cells that we identified in wild-type bone marrow. In vitro, we show that an early IL-7R(+) lymphoid progenitor population expresses IL 3R and proliferates in response to IL-3 and that IL-3 activates Stat5 comparably to IL-7. Clinically, IL-3 may therefore be useful treatment for X-linked and Jak3 deficient SCID patients who lack bone marrow donors. PMID- 10477720 TI - The presence of novel amino acids in the cytoplasmic domain of stem cell factor results in hematopoietic defects in Steel(17H) mice. AB - Stem cell factor (SCF) is expressed as an integral membrane growth factor that may be differentially processed to produce predominantly soluble (S) (SCF(248)) or membrane-associated (MA) (SCF(220)) protein. A critical role for membrane presentation of SCF in the hematopoietic microenvironment (HM) has been suggested from the phenotype of the Steel-dickie (Sl(d)) mice, which lack MA SCF, and by studies performed in our laboratory (and by others) using long-term bone marrow cultures and transgenic mice expressing different SCF isoforms. Steel(17H) (Sl(17H)) is an SCF mutant that demonstrates melanocyte defects and sterility in males but not in females. The Sl(17H) allele contains a intronic mutation resulting in the substitution of 36 amino acids (aa's) in the SCF cytoplasmic domain with 28 novel aa's. This mutation, which affects virtually the entire cytoplasmic domain of SCF, could be expected to alter membrane SCF presentation. To investigate this possibility, we examined the biochemical and biologic properties of the Sl(17H)-encoded protein and its impact in vivo and in vitro on hematopoiesis and on c-Kit signaling. We demonstrate that compound heterozygous Sl/Sl(17H) mice manifest multiple hematopoietic abnormalities in vivo, including red blood cell deficiency, bone marrow hypoplasia, and defective thymopoiesis. In vitro, both S and MA Sl(17H) isoforms of SCF exhibit reduced cell surface expression on stromal cells and diminished biological activity in comparison to wild-type (wt) SCF isoforms. These alterations in presentation and biological activity are associated with a significant reduction in the proliferation of an SCF-responsive erythroid progenitor cell line and in the activation of phosphatidylinositol 3-Kinase/Akt and mitogen-activated protein-Kinase signaling pathways. In vivo, transgene expression of the membrane-restricted (MR) (SCF(X9/D3)) SCF in Sl/Sl(17H) mutants results in a significant improvement in peripheral red blood cell counts in comparison to Sl/Sl(17H) mice. PMID- 10477721 TI - Rapid differentiation of a rare subset of adult human lin(-)CD34(-)CD38(-) cells stimulated by multiple growth factors in vitro. AB - Recently, several reports of lineage-negative (lin(-)) CD34(-) cells with in vivo hematopoietic activity have focused interest on the properties and growth factor response characteristics of these cells. We have now identified a combination of 5 growth factors that are necessary and sufficient to stimulate a marked mitogenic and differentiation response by a subset of human lin(-)CD34(-)CD38(-) cells present in normal adult human marrow and granulocyte colony-stimulating factor (G-CSF)-mobilized blood. Less than 0.1% of the cells in highly purified (including doubly sorted) lin(-)CD34(-)CD38(-) cells from these 2 sources formed colonies directly in semisolid medium or generated such cells after 6 weeks in long-term culture. Nevertheless, approximately 1% of the same lin(-)CD34(-)CD38( ) cells were able to proliferate rapidly in serum-free liquid suspension cultures containing human flt-3 ligand, Steel factor, thrombopoietin, interleukin-3 (IL 3), and hyper-IL-6 to produce a net 28- +/- 8-fold increase in total cells within 10 days. Of the cells present in these 10-day cultures, 5% +/- 2% were CD34(+) and 2.5% +/- 0.9% were erythroid, granulopoietic, megakaryocytopoietic, or multilineage colony-forming cells (CFC) (13 +/- 7 CFC per lin(-)CD34(-)CD38(-) pre-CFC). In contrast to lin(-)CD34(+)CD38(-) cells, this response of lin(-)CD34( )CD38(-) cells required exposure to all of the 5 growth factors used. Up to 1.7 x 10(5) lin(-)CD34(-) adult marrow cells failed to engraft sublethally irradiated NOD/SCID-beta(2)M(-/-) mice. These studies demonstrate unique properties of a rare subset of lin(-)CD34(-)CD38(-) cells present in both adult human marrow and mobilized blood samples that allow their rapid proliferation and differentiation in vitro within an overall period of 3 to 4 weeks. The rapidity of this response challenges current concepts about the normal duration and coordinated control of these processes in adults. PMID- 10477722 TI - Identification of a 14-3-3 binding sequence in the common beta chain of the granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and IL-5 receptors that is serine-phosphorylated by GM-CSF. AB - The common beta chain (beta(c)) of the granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and IL-5 receptors is the major signaling subunit of these receptors coupling ligand binding to multiple biological activities. It is thought that these multiple functions arise as a consequence of the recruitment of specific signaling molecules to tyrosine-phosphorylated residues in the cytoplasmic domain of beta(c). However, the contribution of serine phosphorylation in beta(c) to the recruitment of signaling molecules is not known. We show here the identification of a phosphoserine motif in the cytoplasmic domain of beta(c) that interacts with the adaptor protein 14-3-3zeta. Coimmunoprecipitation and pull-down experiments with a glutathione S-transferase (GST):14-3-3zeta fusion protein showed that 14-3-3 directly associates with beta(c) but not the GM-CSF receptor alpha chain. C-terminal truncation mutants of beta(c) further showed that a region between amino acids 544 and 626 in beta(c) was required for its association with 14-3-3zeta. This region contains the sequence (582)HSRSLP(587), which closely resembles the RSXSXP (where S is phosphorylated) consensus 14-3-3 binding site identified in a number of signaling molecules, including Raf-1. Significantly, substitution of (582)HSRSLP(587) for EFAAAA completely abolished interaction of beta(c) with GST-14-3-3zeta. Furthermore, the interaction of beta(c) with GST-14-3-3 was greatly reduced in the presence of a peptide containing the 14-3-3 binding site, but only when (585)Ser was phosphorylated. Direct binding experiments showed that the peptide containing phosphorylated (585)Ser bound 14-3-3zeta with an affinity of 150 nmol/L. To study the regulation of (585)S phosphorylation in vivo, we raised antibodies that specifically recognized (585)Ser-phosphorylated beta(c). Using these antibodies, we showed that GM-CSF stimulation strongly upregulated (585)Ser phosphorylation in M1 myeloid leukemic cells. The proximity of the SHC-binding site ((577)Tyr) to the 14-3-3-binding site ((582)HSRSLP(587)) and their conservation between mouse, rat, and human beta(c) but not in other cytokine receptors suggest that they form a distinct motif that may subserve specialized functions associated with the GM-CSF, IL-3, and IL-5 receptors. PMID- 10477723 TI - Simultaneous antagonism of interleukin-5, granulocyte-macrophage colony stimulating factor, and interleukin-3 stimulation of human eosinophils by targetting the common cytokine binding site of their receptors. AB - Human interleukin-5 (IL-5), granulocyte-macrophage colony-stimulating factor (GM CSF), and IL-3 are eosinophilopoietic cytokines implicated in allergy in general and in the inflammation of the airways specifically as seen in asthma. All 3 cytokines function through cell surface receptors that comprise a ligand-specific alpha chain and a shared subunit (beta(c)). Although binding of IL-5, GM-CSF, and IL-3 to their respective receptor alpha chains is the first step in receptor activation, it is the recruitment of beta(c) that allows high-affinity binding and signal transduction to proceed. Thus, beta(c) is a valid yet untested target for antiasthma drugs with the added advantage of potentially allowing antagonism of all 3 eosinophil-acting cytokines with a single compound. We show here the first development of such an agent in the form of a monoclonal antibody (MoAb), BION-1, raised against the isolated membrane proximal domain of beta(c). BION-1 blocked eosinophil production, survival, and activation stimulated by IL-5 as well as by GM-CSF and IL-3. Studies of the mechanism of this antagonism showed that BION-1 prevented the high-affinity binding of (125)I-IL-5, (125)I-GM-CSF, and (125)I-IL-3 to purified human eosinophils and that it bound to the major cytokine binding site of beta(c). Interestingly, epitope analysis using several beta(c) mutants showed that BION-1 interacted with residues different from those used by IL-5, GM-CSF, and IL-3. Furthermore, coimmunoprecipitation experiments showed that BION-1 prevented ligand-induced receptor dimerization and phosphorylation of beta(c), suggesting that ligand contact with beta(c) is a prerequisite for recruitment of beta(c), receptor dimerization, and consequent activation. These results demonstrate the feasibility of simultaneously inhibiting IL-5, GM-CSF, and IL-3 function with a single agent and that BION-1 represents a new tool and lead compound with which to identify and generate further agents for the treatment of eosinophil-dependent diseases such as asthma. PMID- 10477724 TI - Production of thrombopoietin by human carcinomas and its novel isoforms. AB - Thrombocytosis is occasionally seen in patients with carcinomas and has been assumed to be attributable to interleukin-6 or granulocyte-macrophage colony stimulating factor produced by carcinoma cells. In this study, we clarified whether thrombopoietin (TPO) is involved in carcinoma-associated thrombocytosis. Expression of TPO mRNA was observed in the majority of 27 carcinoma cell lines as determined by reverse transcriptase-polymerase chain reaction (RT-PCR). There were 6 PCR products differing in size; sequence analysis showed the full-length TPO mRNA (TPO-1), 12- and 116-bp deleted variants (TPO-2 and TPO-3, respectively), and 3 novel isoforms (197- and 128-bp deleted forms and a 60-bp insert form of TPO-3; named TPO-4, TPO-5, and TPO-6, respectively). Of 27 lines, 24 expressed TPO-1 mRNA with various other isoforms. Culture supernatants of COS 1 cells transfected with TPO-5 or TPO-6 cDNA did not promote the proliferation of TPO-responsive cells, whereas Western blot analysis on the cell lysates demonstrated TPO-5 but not TPO-6 protein, suggesting poor extracellular secretion (TPO-5) or poor protein synthesis (TPO-6). TPO protein was detected in 10-fold concentrated culture supernatants of cells of these carcinoma lines, with a median concentration of 0.38 fmol/mL as evaluated by enzyme-linked immunosorbent assay. High blood TPO levels were observed with a median value of 3.46 fmol/mL (range, 0.34 to 8.67 fmol/mL) in patients with advanced carcinomas associated with thrombocytosis. These results indicate that thrombocytosis in patients with carcinomas might be caused, at least in part, by TPO produced by carcinoma cells. PMID- 10477725 TI - Transforming growth factor-beta1 (TGF-beta1) induces thrombopoietin from bone marrow stromal cells, which stimulates the expression of TGF-beta receptor on megakaryocytes and, in turn, renders them susceptible to suppression by TGF-beta itself with high specificity. AB - The present study was designed to test the concept that platelets release a humoral factor that plays a regulatory role in megakaryopoiesis. The results showed that, among various hematoregulatory cytokines examined, transforming growth factor-beta1 (TGF-beta1) was by far the most potent enhancer of mRNA expression of bone marrow stromal thrombopoietin (TPO), a commitment of lineage specificity. The TPO, in turn, induced TGF-beta receptors I and II on megakaryoblasts at the midmegakaryopoietic stage; at this stage, TGF-beta1 was able to arrest the maturation of megakaryocyte colony-forming units (CFU-Meg). This effect was relatively specific when compared with its effect on burst forming unit-erythroid (BFU-E) or colony-forming unit-granulocyte-macrophage (CFU GM). In patients with idiopathic thrombocytopenic purpura (ITP), the levels of both TGF-beta1 and stromal TPO mRNA were correlatively increased and an arrest of megakaryocyte maturation was observed. These in vivo findings are in accord with the aforementioned in vitro results. Thus, the results of the present investigation suggest that TGF-beta1 is one of the pathophysiological feedback regulators of megakaryopoiesis. PMID- 10477726 TI - Involvement of the retinoblastoma protein in monocytic and neutrophilic lineage commitment of human bone marrow progenitor cells. AB - The retinoblastoma gene product (pRb) is involved in both cell cycle regulation and cell differentiation. pRb may have dual functions during cell differentiation: partly by promoting a cell cycle brake at G(1) and also by interacting with tissue-specific transcription factors. We recently showed that pRb mediates differentiation of leukemic cell lines involving mechanisms other than the induction of G(1) arrest. In the present study, we investigated the role of pRb in differentiation of human bone marrow progenitor cells. Human bone marrow cells were cultured in a colony-forming unit-granulocyte-macrophage (CFU GM) assay. The addition of antisense RB oligonucleotides (alpha-RB), but not the addition of sense orientated oligonucleotides (SO) or scrambled oligonucleotides (SCR), reduced the number of colonies staining for nonspecific esterase without affecting the clonogenic growth. Monocytic differentiation of CD34(+) cells supported by FLT3-ligand and interleukin-3 (IL-3) was correlated to high levels of hypophosphorylated pRb, whereas neutrophilic differentiation, supported by granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF), was correlated to low levels. The addition of alpha-RB to liquid cultures of CD34(+) cells, supported with FLT3-ligand and IL-3, inhibited monocytic differentiation. This was judged by morphology, the expression of CD14, and staining for esterase. Moreover, the inhibition of monocytic differentiation of CD34(+) cells mediated by alpha-RB, which is capable of reducing pRb expression, was counterbalanced by an enhanced neutrophilic differentiation response, as judged by morphology and the expression of lactoferrin. CD34(+) cells incubated with oligo buffer, alpha RB, SO, or SCR showed similar growth rates. Taken together, these data suggest that pRb plays a critical role in the monocytic and neutrophilic lineage commitment of human bone marrow progenitors, probably by mechanisms that are not strictly related to control of cell cycle progression. PMID- 10477727 TI - Signaling via Src family kinases is required for normal internalization of the receptor c-Kit. AB - Stem cell factor (SCF) exerts its biological effects by binding to a specific receptor, the tyrosine kinase c-Kit, which is expressed on the cell surface. Although normal cellular trafficking of growth factor receptors may play a critical role in the modulation of receptor function, the mechanisms that regulate the distribution of c-Kit on the cell surface and the internalization of c-Kit have not been fully defined. We investigated whether signal transduction via Src family kinases is required for normal c-Kit trafficking. Treatment of the SCF-responsive human hematopoietic cell line MO7e with the inhibitor of Src family kinases PP1 blocked SCF-induced capping of c-Kit and internalization of c Kit. c-Kit was able to associate with clathrin in the presence of PP1, suggesting that entry of c-Kit into clathrin-coated pits occurs independently of Src family kinases. SCF-induced internalization of c-Kit was also diminished in the D33-3 lymphoid cell line in which expression of Lyn kinase was disrupted by homologous recombination. These results indicate that Src family kinases play a role in ligand-induced trafficking of c-Kit. PMID- 10477728 TI - Osteoclast-derived zinc finger (OCZF) protein with POZ domain, a possible transcriptional repressor, is involved in osteoclastogenesis. AB - The differentiation of osteoclasts is regulated by transcription factors expressed in cells of osteoclast lineage. We isolated here a potential transcription factor from a cDNA library of an enriched population of preosteoclasts and osteoclasts. The cDNA encodes a protein with N-terminal POZ domain and C-terminal Kruppel-like zinc fingers. We designate this protein as osteoclast-derived zinc finger (OCZF). OCZF was found to be rat homologue of mouse leukemia/lymphoma-related factor (LRF). Northern blot and in situ hybridization analysis showed OCZF mRNA at a high level in osteoclasts and kidney cells. OCZF had a nuclear targeting sequence and was localized in the nucleus of transfected cells. In addition, OCZF specifically bound to the guanine-rich consensus sequences of Egr-1 and c-Krox. Transient transfection assays indicate that OCZF can repress transcription activity like other POZ domain proteins. Furthermore, antisense but not sense phosphorothioate oligodeoxynucleotides (ODNs) for OCZF cDNA suppressed the formation of osteoclast-like multinucleated cells (MNCs) in bone marrow culture, whereas the same ODNs did not significantly affect the formation of macrophage polykaryons and mononuclear preosteoclast-like cells (POCs). These results suggest that OCZF is a unique transcription factor that plays an important role in the late stage of osteoclastogenesis. PMID- 10477729 TI - ATM is upregulated during the mitogenic response in peripheral blood mononuclear cells. AB - Patients with the human genetic disorder ataxia-telangiectasia (A-T) are characterized by immunodeficiency and a predisposition to develop lymphoid malignancies. The gene mutated in A-T patients, ATM, codes for a high molecular weight protein that is implicated in DNA damage recognition and cell cycle control. The ATM protein does not change in amount or cellular distribution throughout the cell cycle or in response to DNA damaging agents. Because peripheral blood mononuclear cells (PBMCs) are largely in a state of quiescence and can be readily stimulated to enter a proliferative phase and because A-T cells exhibit growth abnormalities and senescence, indicative of a general intracellular defect in signalling, we chose PBMCs to examine the relationship of ATM to the proliferative status of the cell. We show here that ATM protein is present at low levels in freshly isolated PBMCs and increases approximately 6 fold to 10-fold in response to a mitogenic stimulus, reaching a maximum after 3 to 4 days. A similar, but delayed response, was evident in the presence of serum only. This increase in ATM protein was accompanied by an increase in ATM kinase activity. While expression of ATM protein increased during proliferation, ATM mRNA expression was unchanged in stimulated and unstimulated cells and there was no evidence for increased ATM protein stability in the phytohemagglutinin (PHA) treated cells. In keeping with the reduced levels of ATM in quiescent cells, the extent of radiation-induction of the p53 pathway was significantly lower than in mitogen-stimulated cells. Basal levels of p21 were elevated in quiescent cells, and the response to radiation was negligible or reduced compared with proliferating cells over a 2-hour period. Overall, the data suggest that the increase in ATM protein in proliferating cells is due to posttranscriptional regulation and points to a role for ATM in more general signalling. PMID- 10477730 TI - Defensin promotes the binding of lipoprotein(a) to vascular matrix. AB - Retention of lipoproteins within the vasculature is a central event in the pathogenesis of atherosclerosis. However, the signals that mediate this process are only partially understood. Prompted by putative links between inflammation and atherosclerosis, we previously reported that alpha-defensins released by neutrophils are present in human atherosclerotic lesions and promote the binding of lipoprotein(a) [Lp(a)] to vascular cells without a concomitant increase in degradation. We have now tested the hypothesis that this accumulation results from the propensity of defensin to form stable complexes with Lp(a) that divert the lipoprotein from its normal cellular degradative pathways to the extracellular matrix (ECM). In accord with this hypothesis, defensin stimulated the binding of Lp(a) to vascular matrices approximately 40-fold and binding of the reactants to the matrix was essentially irreversible. Defensin formed stable, multivalent complexes with Lp(a) and with its components, apoprotein (a) and low density lipoprotein (LDL), as assessed by optical biosensor analysis, gel filtration, and immunoelectron microscopy. Binding of defensin/Lp(a) complexes to matrix was inhibited (>90%) by heparin and by antibodies to fibronectin (>70%), but not by antibodies to vitronectin or thrombospondin. Defensin increased the binding of Lp(a) (10 nmol/L) to purified fibronectin more than 30-fold. Whereas defensin and Lp(a) readily traversed the endothelial cell membranes individually, defensin/Lp(a) complexes lodged on the cell surface. These studies demonstrate that alpha-defensins released from activated or senescent neutrophils stimulate the binding of an atherogenic lipoprotein to the ECM of endothelial cells, a process that may contribute to lipoprotein accumulation in atherosclerotic lesions. PMID- 10477732 TI - Fibrin fragment induction of plasminogen activator inhibitor transcription is mediated by activator protein-1 through a highly conserved element. AB - Plasminogen activator inhibitor type-1 (PAI-1), a serine protease inhibitor, affects the processes of fibrinolysis, wound healing, and vascular remodeling. We have demonstrated that PAI-1 transcription is induced by D dimer, a plasmin proteolytic fragment of fibrin, supporting its role in negative feedback on peri cellular proteolysis. The focus of this study was to define the mechanism of D dimer's effects on PAI-1 transcription. D dimer increased the binding activity of the transcription factor activator protein-1 components c-fos/junD and c-fos mRNA levels in a time- and concentration-dependent manner to a greater extent than fibrinogen. Both basal and D dimer-induced PAI-1 transcriptional activity were entirely dependent on elements within the -161 to -48 bp region of the PAI-1 gene in fibroblasts. Mutations within the AP-1-like element (-59 to -52 bp) in the PAI 1 gene affected D dimer-induced transcriptional activity, c-fos/junD DNA binding, and basal and c-fos inducible PAI-1 transcriptional activity. Furthermore, expression of either wild-type or mutant c-fos proteins augmented or diminished the response of the PAI-1 promoter (-161 to +26 bp) to D dimer, respectively. D dimer-induced binding of c-fos/junD to the highly conserved and unique AP-1 like element in the PAI-1 gene provides a mechanism whereby specific fibrin fragments control fibrin persistence at sites of inflammation, fibrosis, and neoplasia. PMID- 10477731 TI - Cultured endothelial cells from human arteriovenous malformations have defective growth regulation. AB - Vascular malformations are frequent in newborns, and they persist throughout life, which differentiates them from vascular tumors (eg, hemangiomas). Arteriovenous malformations are high-flow vascular malformations. They are considered nonmalignant but can expand and become a significant clinical risk when extensive. To characterize endothelial cells from arteriovenous malformations (AMEC), we cultured cells obtained from surgical specimens and studied their properties. After selection, the cells that grew out from explants had phenotypic and antigenic features (platelet endothelial cell adhesion molecule, von Willebrand factor) of human endothelial cells. Their spontaneous proliferation rate was higher (1.8 to 6.4 times) than that of human umbilical vein, arterial, or microvascular endothelial cells. The proliferation rate of AMEC was not sensitive to the inhibitory activity of various cytokines (interleukin-1beta, tumor necrosis factor-alpha, transforming growth factor-beta, Interferon-gamma). In basal conditions, intercellular adhesion molecule (ICAM-1) was detected at a higher level of expression (6- to 10-fold) on AMEC, but these cells failed to express E-selectin or the vascular cell adhesion molecule (VCAM 1) after cytokine stimulation. Expression of c-ets-1 proto-oncogene was shown by in situ hybridization. The low response to cytokines, the higher propensity to proliferate, and the ets-1 expression suggest that AMEC have a defective regulation of proliferation that may be due to a reduced apoptotic process. PMID- 10477733 TI - The human platelet alphaIIb gene is not closely linked to its integrin partner beta3. AB - alphaIIbb3 integrin is a heterodimeric receptor facilitating platelet aggregation. Both genes are on chromosome 17q21.32. Intergenic distance between them has been reported to be 125 to 260 kilobasepairs (kb) by pulsed-field gel electrophoresis (PFGE) genomic analysis, suggesting that they may be regulated coordinately during megakaryopoiesis. In contrast, other studies suggest these genes are greater than 2.0 megabasepairs (mb) apart. Because of the potential biological implications of having these two megakaryocytic-specific genes contiguous, we attempted to resolve this discrepancy. Taking advantage of large kindreds with mutations in either alphaIIb or beta3, we have developed a genetic linkage map between the thyroid receptor hormone-1 gene (THRA1) and beta3 as follows: cen-THRA1-BRCA1-D17S579/alphaIIb-beta3-qte r, with a distance of 1.3 centiMorgans (cM) between alphaIIb and beta3 and the two genes being oriented in the same direction. PFGE genomic and YAC clone analysis showed that the beta3 gene is distal and >/=365 kb upstream of alphaIIb. Additional restriction mapping shows alphaIIb is linked to the erythrocyte band 3 (EPB3) gene, and beta3 to the homeobox HOX2b gene. Analysis of alphaIIb(+)-BAC and P1 clones confirm that the EPB3 gene is approximately 110 kb downstream of the alphaIIb gene. Sequencing the region surrounding the human alphaIIb locus showed the Granulin gene approximately 18 kb downstream to alphaIIb, and the KIAA0553 gene approximately 5.7 kb upstream. This organization is conserved in the murine sequence. These studies show that alphaIIb and beta3 are not closely linked, with alphaIIb flanked by nonmegakaryocytic genes, and imply that they are unlikely to share common regulatory domains during megakaryopoiesis. PMID- 10477734 TI - CD34(+) acute myeloid and lymphoid leukemic blasts can be induced to differentiate into dendritic cells. AB - CD34(+) hematopoietic stem cells from normal individuals and from patients with chronic myelogenous leukemia can be induced to differentiate into dendritic cells (DC). The aim of the current study was to determine whether acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) cells could be induced to differentiate into DC. CD34(+) AML-M2 cells with chromosome 7 monosomy were cultured in the presence of granulocyte-macrophage colony-stimulating factor (GM CSF), tumor necrosis factor alpha (TNFalpha), and interleukin-4 (IL-4). After 3 weeks of culture, 35% of the AML-M2 cells showed DC morphology and phenotype. The DC phenotype was defined as upmodulation of the costimulatory molecules CD80 and CD86 and the expression of CD1a or CD83. The leukemic nature of the DC was validated by detection of chromosome 7 monosomy in sorted DC populations by fluorescence in situ hybridization (FISH). CD34(+) leukemic cells from 2 B-ALL patients with the Philadelphia chromosome were similarly cultured, but in the presence of CD40-ligand and IL-4. After 4 days of culture, more than 58% of the ALL cells showed DC morphology and phenotype. The leukemic nature of the DC was validated by detection of the bcr-abl fusion gene in sorted DC populations by FISH. In functional studies, the leukemic DC were highly superior to the parental leukemic blasts for inducing allogeneic T-cell responses. Thus, CD34(+) AML and ALL cells can be induced to differentiate into leukemic DC with morphologic, phenotypic, and functional similarities to normal DC. PMID- 10477735 TI - Isolation of a highly quiescent subpopulation of primitive leukemic cells in chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is characterized by an increased proliferative activity of the leukemic progenitors that produce an elevated number of mature granulocytes. Nevertheless, cell cycle-active agents, even in very high doses, are alone unable to eradicate the leukemic clone, suggesting the presence of a rare subset of quiescent leukemic stem cells. To isolate such cells, we first used Hoechst 33342 and Pyronin Y staining to obtain viable G(0) and G(1)/S/G(2)/M fractions of CD34(+) cells by fluorescence-activated cell sorting (FACS) from 6 chronic-phase CML patients' samples and confirmed the quiescent and cycling status of the 2 fractions by demonstration of expected patterns of Ki-67 and D cyclin expression. Leukemic (Ph(+)/BCR-ABL(+)) cells with in vitro progenitor activity and capable of engrafting immunodeficient mice were identified in the directly isolated G(0) cells. Single-cell reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that many leukemic CD34(+) G(0) cells also expressed BCR-ABL mRNA. CD34(+) from 8 CML patients were also labeled with carboxyfluorescein diacetate succinimidyl diester (CFSE) before being cultured (with and without added growth factors) to allow viable cells that had remained quiescent (ie, CFSE(+)) after 4 days to be retrieved by FACS. Leukemic progenitors were again detected in all quiescent populations isolated by this second strategy, including those exposed to a combination of flt3-ligand, Steel factor, interleukin-3, interleukin-6, and granulocyte colony-stimulating factor. These findings provide the first direct and definitive evidence of a deeply but reversibly quiescent subpopulation of leukemic cells in patients with CML with both in vitro and in vivo stem cell properties. PMID- 10477736 TI - Hodgkin/Reed-Sternberg cells induce fibroblasts to secrete eotaxin, a potent chemoattractant for T cells and eosinophils. AB - Hodgkin's disease is histopathologically characterized by the relative scarcity of neoplastic Hodgkin and Reed-Sternberg cells and for yet unknown reasons by an abundant reactive background of T lymphocytes and often eosinophils. Eotaxin is a CC-chemokine attracting eosinophils and T helper 2 (Th2) cells in allergic inflammation. We now report that eotaxin is strongly expressed in fibroblasts of Hodgkin's disease tissues, whereas Hodgkin/Reed-Sternberg cells do not express this chemokine. In tissue culture, Hodgkin's disease tumor cells induce eotaxin expression in cocultured dermal fibroblasts in a concentration leading to a specific chemotactic response of a Th2 cell clone. Production of tumor necrosis factor-alpha (TNF-alpha) by Hodgkin/Reed-Sternberg cells appears to be responsible for this induction, because blocking of TNF-alpha by neutralizing antibodies prevented fibroblast eotaxin expression. Our data suggest that eotaxin is involved in the pathobiology of Hodgkin's disease by contributing to eosinophil and T-lymphocyte recruitment. PMID- 10477737 TI - The (4;11)(q21;p15) translocation fuses the NUP98 and RAP1GDS1 genes and is recurrent in T-cell acute lymphocytic leukemia. AB - We determined the breakpoint genes of the translocation t(4;11)(q21;p15) that occurred in a case of adult T-cell acute lymphocytic leukemia (T-ALL). The chromosome 11 breakpoint was mapped to the region between D11S470 and D11S860. The nucleoporin 98 gene (NUP98), which is rearranged in several acute myeloid leukemia translocations, is located within this region. Analysis of somatic cell hybrids segregating the translocation chromosomes showed that the chromosome 11 breakpoint occurs within NUP98. The fusion partner of NUP98 was identified as the RAP1GDS1 gene using 3' RACE. RAP1GDS1 codes for smgGDS, a ubiquitously expressed guanine nucleotide exchange factor that stimulates the conversion of the inactive GDP-bound form of several ras family small GTPases to the active GTP-bound form. In the NUP98-RAP1GDS1 fusion transcript (abbreviated as NRG), the 5' end of the NUP98 gene is joined in frame to the coding region of the RAP1GDS1 gene. This joins the FG repeat-rich region of NUP98 to RAP1GDS1, which largely consists of tandem armadillo repeats. NRG fusion transcripts were detected in the leukemic cells of 2 other adult T-ALL patients. One of these patients had a variant translocation with a more 5' breakpoint in NUP98. This is the first report of an NUP98 translocation in lymphocytic leukemia and the first time that RAP1GDS1 has been implicated in any human malignancy. PMID- 10477738 TI - Interleukin-6 regulation of matrix metalloproteinase (MMP-2 and MMP-9) and tissue inhibitor of metalloproteinase (TIMP-1) expression in malignant non-Hodgkin's lymphomas. AB - We showed previously that human malignant non-Hodgkin's lymphomas (NHL) degrade extracellular matrix (ECM) components through the action of metalloproteinases and that elevated expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) correlated with a poor clinical outcome in patients with NHL. In the present study we sought to investigate whether there is any correlation between the expression of gelatinases (MMP-2 and MMP-9), TIMP 1, and the expression of cytokines and growth factors such as interleukin-1beta (IL-1beta), IL-6, IL-10, tumor necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGFbeta), and basic fibroblast growth factor (bFGF) in human NHL. In lymphoma tissues obtained from 32 patients, elevated expression of IL-6 correlated significantly with elevated messenger RNA (mRNA) levels of MMP-9, MMP 2, and TIMP-1. Moreover, in human lymphoid cell lines of B- and T-cell origin (Raji, Jurkat, and NC-37), IL-6 stimulated production of MMP-9 and MMP-2 but not TIMP-1. In the Matrigel invasion assay IL-6 significantly upregulated transmigration of Raji and Jurkat cells, which in turn was inhibited by recombinant human TIMP-1 and anti-MMP-9 and MMP-2 antibodies. We postulate that IL-6 may play a role in the clinical aggressiveness of human NHL by stimulating MMP production. PMID- 10477739 TI - A novel Epstein-Barr virus-like virus, HV(MNE), in a Macaca nemestrina with mycosis fungoides. AB - Epstein-Barr virus (EBV) infection of humans has been associated with the development of lymphoid malignancies mainly of B-cell lineage, although occasionally T-cell lymphomas have been reported. We describe here the characterization of a novel EBV-like virus (HV(MNE)) isolated from a simian T cell lymphotropic virus type I/II (STLV-I/II) seronegative pigtailed macaque (Macaca nemestrina) with a cutaneous T-cell lymphoma. Immunohistochemistry studies on the skin lesions demonstrated that the infiltrating cells were of the CD3(+)/CD8(+) phenotype. Two primary transformed CD8(+) T-cell lines were obtained from cultures of peripheral blood mononuclear cells (PBMC) and skin, and, with time, both cell lines became interleukin-2-independent and acquired the constitutive activation of STAT proteins. Polymerase chain reaction analysis of the DNA from the cell lines and tissues from the lymphomatous animal demonstrated the presence of a 536-bp DNA fragment that was 90% identical to EBV polymerase gene sequences, whereas the same DNA was consistently negative for STLV-I/II sequences. Electron microscopy performed on both cell lines, after sodium butyrate treatment, showed the presence of a herpes-like virus that was designated HV(MNE) according to the existing nomenclature. In situ hybridization studies using EBV Epstein-Barr viral-encoded RNA probes showed viral RNA expression in both CD8(+) T-cell lines as well as in the infiltrating CD8(+) T cells of skin-tissue biopsies. Phylogenetic analysis of a 465-bp fragment from the polymerase gene of HV(MNE) placed this virus within the Lymphocryptovirus genus and demonstrated that HV(MNE) is a distinct virus, clearly related to human EBV and other EBV-like herpesviruses found in nonhuman primates. PMID- 10477740 TI - Arsenic trioxide selectively induces acute promyelocytic leukemia cell apoptosis via a hydrogen peroxide-dependent pathway. AB - Low concentrations of As(2)O(3) (/=75% decrease, while higher concentrations caused >/=96% decrease in proliferation. Loss of PBMC proliferation was not due to either mPEG-induced cytotoxicity, as viability was normal, or cellular anergy, as phytohemagglutinin (PHA)-stimulated mPEG-PBMC demonstrated normal proliferative responses. Addition of exogenous interleukin (IL)-2 also had no proliferative effect, suggesting that the mPEG-modified T cells were not antigen primed. Flow cytometric analysis demonstrates that mPEG modification dramatically decreases antibody recognition of multiple molecules involved in essential cell:cell interactions, including both T-cell molecules (CD2, CD3, CD4, CD8, CD28, CD11a, CD62L) and antigen-presenting cell (APC) molecules (CD80, CD58, CD62L) likely preventing the initial adhesion and costimulatory events necessary for immune recognition and response. PMID- 10477745 TI - Behavior and therapeutic efficacy of beta-glucuronidase-positive mononuclear phagocytes in a murine model of mucopolysaccharidosis type VII. AB - Bone marrow transplantation (BMT) is relatively effective for the treatment of lysosomal storage diseases. To better understand the contribution of specific hematopoietic lineages to the efficacy of BMT, we transplanted beta-glucuronidase positive mononuclear phagocytes derived from either the peritoneum or from bone marrow in vitro into syngeneic recipients with mucopolysaccharidosis type VII (MPS VII). Cell surface marking studies indicate that the bone marrow-derived cells are less mature than the peritoneal macrophages. However, both cell types retain the ability to home to tissues rich in cells of the reticuloendothelial system after intravenous injection into MPS VII mice. The half-life of both types of donor macrophages is approximately 7 days, and some cells persist for at least 30 days. In several tissues, therapeutic levels of beta-glucuronidase are present, and histopathologic analysis demonstrates that lysosomal storage is dramatically reduced in the liver and spleen. Macrophages intravenously injected into newborn MPS VII mice localize to the same tissues as adult mice but are also observed in the meninges and parenchyma of the brain. These data suggest that macrophages play a significant role in the therapeutic efficacy of BMT for lysosomal storage diseases and may have implications for treatments such as gene therapy. PMID- 10477746 TI - In vivo selection of wild-type hematopoietic stem cells in a murine model of Fanconi anemia. AB - Fanconi anemia (FA) is an autosomal recessive disorder characterized by birth defects, increased incidence of malignancy, and progressive bone marrow failure. Bone marrow transplantation is therapeutic and, therefore, FA is a candidate disease for hematopoietic gene therapy. The frequent finding of somatic mosaicism in blood of FA patients has raised the question of whether wild-type bone marrow may have a selective growth advantage. To test this hypothesis, a cohort radio ablated wild-type mice were transplanted with a 1:1 mixture of FA group C knockout (FACKO) and wild-type bone marrow. Analysis of peripheral blood at 1 month posttransplantation showed only a moderate advantage for wild-type cells, but upon serial transplantation, clear selection was observed. Next, a cohort of FACKO mice received a transplant of wild-type marrow cells without prior radio ablation. No wild-type cells were detected in peripheral blood after transplantation, but a single injection of mitomycin C (MMC) resulted in an increase to greater than 25% of wild-type DNA. Serial transplantation showed that the selection occurred at the level of hematopoietic stem cells. No systemic side effects were observed. Our results show that in vivo selection for wild-type hematopoietic stem cells occurs in FA and that it is enhanced by MMC administration. PMID- 10477747 TI - Megabase chromatin domains involved in DNA double-strand breaks in vivo. AB - The loss of chromosomal integrity from DNA double-strand breaks introduced into mammalian cells by ionizing radiation results in the specific phosphorylation of histone H2AX on serine residue 139, yielding a specific modified form named gamma H2AX. An antibody prepared to the unique region of human gamma-H2AX shows that H2AX homologues are phosphorylated not only in irradiated mammalian cells but also in irradiated cells from other species, including Xenopus laevis, Drosophila melanogaster, and Saccharomyces cerevisiae. The antibody reveals that gamma-H2AX appears as discrete nuclear foci within 1 min after exposure of cells to ionizing radiation. The numbers of these foci are comparable to the numbers of induced DNA double-strand breaks. When DNA double-strand breaks are introduced into specific partial nuclear volumes of cells by means of a pulsed microbeam laser, gamma-H2AX foci form at these sites. In mitotic cells from cultures exposed to nonlethal amounts of ionizing radiation, gamma-H2AX foci form band-like structures on chromosome arms and on the end of broken arms. These results offer direct visual confirmation that gamma-H2AX forms en masse at chromosomal sites of DNA double strand breaks. The results further suggest the possible existence of units of higher order chromatin structure involved in monitoring DNA integrity. PMID- 10477748 TI - The 193-kD vault protein, VPARP, is a novel poly(ADP-ribose) polymerase. AB - Mammalian vaults are ribonucleoprotein (RNP) complexes, composed of a small ribonucleic acid and three proteins of 100, 193, and 240 kD in size. The 100-kD major vault protein (MVP) accounts for >70% of the particle mass. We have identified the 193-kD vault protein by its interaction with the MVP in a yeast two-hybrid screen and confirmed its identity by peptide sequence analysis. Analysis of the protein sequence revealed a region of approximately 350 amino acids that shares 28% identity with the catalytic domain of poly(ADP-ribose) polymerase (PARP). PARP is a nuclear protein that catalyzes the formation of ADP ribose polymers in response to DNA damage. The catalytic domain of p193 was expressed and purified from bacterial extracts. Like PARP, this domain is capable of catalyzing a poly(ADP-ribosyl)ation reaction; thus, the 193-kD protein is a new PARP. Purified vaults also contain the poly(ADP-ribosyl)ation activity, indicating that the assembled particle retains enzymatic activity. Furthermore, we show that one substrate for this vault-associated PARP activity is the MVP. Immunofluorescence and biochemical data reveal that p193 protein is not entirely associated with the vault particle, suggesting that it may interact with other protein(s). A portion of p193 is nuclear and localizes to the mitotic spindle. PMID- 10477749 TI - A nuclear action of the eukaryotic cochaperone RAP46 in downregulation of glucocorticoid receptor activity. AB - RAP46 is a eukaryotic cochaperone that associates with several proteins, including the heat shock protein hsp70/hsc70 and the glucocorticoid receptor (GR). Here we show a downregulation of GR-mediated transactivation by RAP46 via a mechanism independent of a cytoplasmic action of this cochaperone. We demonstrate a specific cytoplasmic-nuclear recruitment of RAP46 by the liganded GR that results in inhibition of the transactivation function of the receptor. A repeated sequence motif [EEX(4)](8) at the NH(2) terminus of RAP46 or BAG-1L, a larger isoform of RAP46, is responsible for this downregulation of GR activity. BAG-1, a shorter isoform with only a duplication of the [EEX(4)] sequence, does not inhibit GR activity. The [EEX(4)](8) motif, when linked to an otherwise unrelated protein, abrogated the inhibitory action of endogenous RAP46 on GR-mediated transactivation. The nuclear effects of RAP46 and BAG-1L are specific since GR mediated inhibition of AP-1 activity was not affected. These studies identify the [EEX(4)](8) sequence as a signature motif for inhibition of GR-mediated transactivation and demonstrate a specific nuclear action of a eukaryotic cochaperone in the regulation of GR activity. PMID- 10477751 TI - Depolarization and neurotrophins converge on the phosphatidylinositol 3-kinase Akt pathway to synergistically regulate neuronal survival. AB - In this report, we have examined the mechanisms whereby neurotrophins and neural activity coordinately regulate neuronal survival, focussing on sympathetic neurons, which require target-derived NGF and neural activity for survival during development. When sympathetic neurons were maintained in suboptimal concentrations of NGF, coincident depolarization with concentrations of KCl that on their own had no survival effect, synergistically enhanced survival. Biochemical analysis revealed that depolarization was sufficient to activate a Ras-phosphatidylinositol 3-kinase-Akt pathway (Ras-PI3-kinase-Akt), and function blocking experiments using recombinant adenovirus indicated that this pathway was essential for approximately 50% of depolarization-mediated neuronal survival. At concentrations of NGF and KCl that promoted synergistic survival, these two stimuli converged to promote increased PI3-kinase-dependent Akt phosphorylation. This convergent PI3-kinase-Akt pathway was essential for synergistic survival. In contrast, inhibition of calcium/calmodulin-dependent protein kinase II revealed that, while this molecule was essential for depolarization-induced survival, it had no role in KCl- induced Akt phosphorylation, nor was it important for synergistic survival by NGF and KCl. Thus, NGF and depolarization together mediate survival of sympathetic neurons via intracellular convergence on a Ras PI3-kinase-Akt pathway. This convergent regulation of Akt may provide a general mechanism for coordinating the effects of growth factors and neural activity on neuronal survival throughout the nervous system. PMID- 10477750 TI - Human BUBR1 is a mitotic checkpoint kinase that monitors CENP-E functions at kinetochores and binds the cyclosome/APC. AB - Human cells express two kinases that are related to the yeast mitotic checkpoint kinase BUB1. hBUB1 and hBUBR1 bind to kinetochores where they are postulated to be components of the mitotic checkpoint that monitors kinetochore activities to determine if chromosomes have achieved alignment at the spindle equator (Jablonski, S.A., G.K.T. Chan, C.A. Cooke, W.C. Earnshaw, and T.J. Yen. 1998. Chromosoma. 107:386-396). In support of this, hBUB1 and the homologous mouse BUB1 have been shown to be important for the mitotic checkpoint (Cahill, D.P., C. Lengauer, J. Yu, G.J. Riggins, J.K. Willson, S.D. Markowitz, K.W. Kinzler, and B. Vogelstein. 1998. Nature. 392:300-303; Taylor, S.S., and F. McKeon. 1997. Cell. 89:727-735). We now demonstrate that hBUBR1 is also an essential component of the mitotic checkpoint. hBUBR1 is required by cells that are exposed to microtubule inhibitors to arrest in mitosis. Additionally, hBUBR1 is essential for normal mitotic progression as it prevents cells from prematurely entering anaphase. We establish that one of hBUBR1's checkpoint functions is to monitor kinetochore activities that depend on the kinetochore motor CENP-E. hBUBR1 is expressed throughout the cell cycle, but its kinase activity is detected after cells have entered mitosis. hBUBR1 kinase activity was rapidly stimulated when the spindle was disrupted in mitotic cells. Finally, hBUBR1 was associated with the cyclosome/anaphase-promoting complex (APC) in mitotically arrested cells but not in interphase cells. The combined data indicate that hBUBR1 can potentially provide two checkpoint functions by monitoring CENP-E-dependent activities at the kinetochore and regulating cyclosome/APC activity. PMID- 10477752 TI - H-Ras activation promotes cytoplasmic accumulation and phosphoinositide 3-OH kinase association of beta-catenin in epidermal keratinocytes. AB - The mechanisms underlying downregulation of the cadherin/catenin complexes and beta-catenin signaling during tumor progression are not fully understood. We have analyzed the effect of oncogenic H-Ras on E-cadherin/catenin complex formation/stabilization and beta-catenin distribution in epidermal keratinocytes. Microinjection or stable expression of V12Ras into keratinocytes promotes the loss of E-cadherin and alpha-catenin and relocalization of beta-catenin to the cytoplasm and nucleus. Moreover, these effects are dependent on PI3K (phosphoinositide 3-OH kinase) activity. Interestingly, a strong association of p85alpha and p110alpha subunits of PI3K with beta-catenin is induced in V12Ras expressing keratinocytes, and in vitro binding assays show a direct interaction between beta-catenin and p85alpha. Overexpression of either V12Ras or constitutively active p110alpha induces metabolic stabilization of beta-catenin and promotes its accumulation in cytoplasmic and nuclear pools. In addition, the interaction of beta-catenin with the adenomatous polyposis coli protein is blocked in V12Ras and p110alpha transformants though no changes in glycogen synthase kinase 3 beta activity could be detected. Nevertheless, in V12Ras transformants the in vivo phosphorylation of beta-catenin in Ser residues is strongly decreased. These results indicate that H-Ras activation induces the relocalization and cytoplasmic stabilization of beta-catenin by a mechanism involving its interaction with PI3K. PMID- 10477753 TI - Parameters that specify the timing of cytokinesis. AB - One model for the timing of cytokinesis is based on findings that p34(cdc2) can phosphorylate myosin regulatory light chain (LC20) on inhibitory sites (serines 1 and 2) in vitro (Satterwhite, L.L., M.H. Lohka, K.L. Wilson, T.Y. Scherson, L.J. Cisek, J.L. Corden, and T.D. Pollard. 1992. J. Cell Biol. 118:595-605), and this inhibition is proposed to delay cytokinesis until p34(cdc2) activity falls at anaphase. We have characterized previously several kinase activities associated with the isolated cortical cytoskeleton of dividing sea urchin embryos (Walker, G.R., C.B. Shuster, and D.R. Burgess. 1997. J. Cell Sci. 110:1373-1386). Among these kinases and substrates is p34(cdc2) and LC20. In comparison with whole cell activity, cortical H1 kinase activity is delayed, with maximum levels in cortices prepared from late anaphase/telophase embryos. To determine whether cortical associated p34(cdc2) influences cortical myosin II activity during cytokinesis, we labeled eggs in vivo with [(32)P]orthophosphate, prepared cortices, and mapped LC20 phosphorylation through the first cell division. We found no evidence of serine 1,2 phosphorylation at any time during mitosis on LC20 from cortically associated myosin. Instead, we observed a sharp rise in serine 19 phosphorylation during anaphase and telophase, consistent with an activating phosphorylation by myosin light chain kinase. However, serine 1,2 phosphorylation was detected on light chains from detergent-soluble myosin II. Furthermore, cells arrested in mitosis by microinjection of nondegradable cyclin B could be induced to form cleavage furrows if the spindle poles were physically placed in close proximity to the cortex. These results suggest that factors independent of myosin II inactivation, such as the delivery of the cleavage stimulus to the cortex, determine the timing of cytokinesis. PMID- 10477754 TI - Gamma-synergin: an EH domain-containing protein that interacts with gamma adaptin. AB - The AP-1 adaptor complex is associated with the TGN, where it links selected membrane proteins to the clathrin lattice, enabling these proteins to be concentrated in clathrin-coated vesicles. To identify other proteins that participate in the clathrin-coated vesicle cycle at the TGN, we have carried out a yeast two- hybrid library screen using the gamma-adaptin subunit of the AP-1 complex as bait. Two novel, ubiquitously expressed proteins were found: p34, which interacts with both gamma-adaptin and alpha-adaptin, and gamma-synergin, an alternatively spliced protein with an apparent molecular mass of approximately 110-190 kD, which only interacts with gamma-adaptin. gamma-Synergin is associated with AP-1 both in the cytosol and on TGN membranes, and it is strongly enriched in clathrin-coated vesicles. It binds directly to the ear domain of gamma-adaptin and it contains an Eps15 homology (EH) domain, although the EH domain is not part of the gamma-adaptin binding site. In cells expressing alpha-adaptin with the gamma-adaptin ear, a construct that goes mainly to the plasma membrane, much of the gamma-synergin is also rerouted to the plasma membrane, indicating that it follows AP-1 onto membranes rather than leading it there. The presence of an EH domain suggests that gamma-synergin links the AP-1 complex to another protein or proteins. PMID- 10477755 TI - mini spindles: A gene encoding a conserved microtubule-associated protein required for the integrity of the mitotic spindle in Drosophila. AB - We describe a new Drosophila gene, mini spindles (msps) identified in a cytological screen for mitotic mutant. Mutation in msps disrupts the structural integrity of the mitotic spindle, resulting in the formation of one or more small additional spindles in diploid cells. Nucleation of microtubules from centrosomes, metaphase alignment of chromosomes, or the focusing of spindle poles appears much less affected. The msps gene encodes a 227-kD protein with high similarity to the vertebrate microtubule-associated proteins (MAPs), human TOGp and Xenopus XMAP215, and with limited similarity to the Dis1 and STU2 proteins from fission yeast and budding yeast. Consistent with their sequence similarity, Msps protein also associates with microtubules in vitro. In the embryonic division cycles, Msps protein localizes to centrosomal regions at all mitotic stages, and spreads over the spindles during metaphase and anaphase. The absence of centrosomal staining in interphase of the cellularized embryos suggests that the interactions between Msps protein and microtubules or centrosomes may be regulated during the cell cycle. PMID- 10477756 TI - The role of actin in spindle orientation changes during the Saccharomyces cerevisiae cell cycle. AB - In the budding yeast Saccharomyces cerevisiae, the mitotic spindle must align along the mother-bud axis to accurately partition the sister chromatids into daughter cells. Previous studies showed that spindle orientation required both astral microtubules and the actin cytoskeleton. We now report that maintenance of correct spindle orientation does not depend on F-actin during G2/M phase of the cell cycle. Depolymerization of F-actin using Latrunculin-A did not perturb spindle orientation after this stage. Even an early step in spindle orientation, the migration of the spindle pole body (SPB), became actin-independent if it was delayed until late in the cell cycle. Early in the cell cycle, both SPB migration and spindle orientation were very sensitive to perturbation of F-actin. Selective disruption of actin cables using a conditional tropomyosin double-mutant also led to defects in spindle orientation, even though cortical actin patches were still polarized. This suggests that actin cables are important for either guiding astral microtubules into the bud or anchoring them in the bud. In addition, F actin was required early in the cell cycle for the development of the actin independent spindle orientation capability later in the cell cycle. Finally, neither SPB migration nor the switch from actin-dependent to actin-independent spindle behavior required B-type cyclins. PMID- 10477757 TI - Microtubule targeting of substrate contacts promotes their relaxation and dissociation. AB - We recently showed that substrate contact sites in living fibroblasts are specifically targeted by microtubules (Kaverina, I., K. Rottner, and J.V. Small. 1998. J. Cell Biol. 142:181-190). Evidence is now provided that microtubule contact targeting plays a role in the modulation of substrate contact dynamics. The results are derived from spreading and polarized goldfish fibroblasts in which microtubules and contact sites were simultaneously visualized using proteins conjugated with Cy-3, rhodamine, or green fluorescent protein. For cells allowed to spread in the presence of nocodazole the turnover of contacts was retarded, as compared with controls and adhesions that were retained under the cell body were dissociated after microtubule reassembly. In polarized cells, small focal complexes were found at the protruding cell front and larger adhesions, corresponding to focal adhesions, at the retracting flanks and rear. At retracting edges, multiple microtubule contact targeting preceded contact release and cell edge retraction. The same effect could be observed in spread cells, in which microtubules were allowed to reassemble after local disassembly by the application of nocodazole to one cell edge. At the protruding front of polarized cells, focal complexes were also targeted and as a result remained either unchanged in size or, more rarely, were disassembled. Conversely, when contact targeting at the cell front was prevented by freezing microtubule growth with 20 nM taxol and protrusion stimulated by the injection of constitutively active Rac, peripheral focal complexes became abnormally enlarged. We further found that the local application of inhibitors of myosin contractility to cell edges bearing focal adhesions induced the same contact dissociation and edge retraction as observed after microtubule targeting. Our data are consistent with a mechanism whereby microtubules deliver localized doses of relaxing signals to contact sites to retard or reverse their development. We propose that it is via this route that microtubules exert their well-established control on cell polarity. PMID- 10477758 TI - Myosin Va movements in normal and dilute-lethal axons provide support for a dual filament motor complex. AB - To investigate the role that myosin Va plays in axonal transport of organelles, myosin Va-associated organelle movements were monitored in living neurons using microinjected fluorescently labeled antibodies to myosin Va or expression of a green fluorescent protein-myosin Va tail construct. Myosin Va-associated organelles made rapid bi-directional movements in both normal and dilute-lethal (myosin Va null) neurites. In normal neurons, depolymerization of microtubules by nocodazole slowed, but did not stop movement. In contrast, depolymerization of microtubules in dilute-lethal neurons stopped movement. Myosin Va or synaptic vesicle protein 2 (SV2), which partially colocalizes with myosin Va on organelles, did not accumulate in dilute-lethal neuronal cell bodies because of an anterograde bias associated with organelle transport. However, SV2 showed peripheral accumulations in axon regions of dilute-lethal neurons rich in tyrosinated tubulin. This suggests that myosin Va-associated organelles become stranded in regions rich in dynamic microtubule endings. Consistent with these observations, presynaptic terminals of cerebellar granule cells in dilute-lethal mice showed increased cross-sectional area, and had greater numbers of both synaptic and larger SV2 positive vesicles. Together, these results indicate that myosin Va binds to organelles that are transported in axons along microtubules. This is consistent with both actin- and microtubule-based motors being present on these organelles. Although myosin V activity is not necessary for long-range transport in axons, myosin Va activity is necessary for local movement or processing of organelles in regions, such as presynaptic terminals that lack microtubules. PMID- 10477760 TI - Apical spectrin is essential for epithelial morphogenesis but not apicobasal polarity in Drosophila. AB - Changes in cell shape and position drive morphogenesis in epithelia and depend on the polarized nature of its constituent cells. The spectrin-based membrane skeleton is thought to be a key player in the establishment and/or maintenance of cell shape and polarity. We report that apical beta(Heavy)-spectrin (beta(H)), a terminal web protein that is also associated with the zonula adherens, is essential for normal epithelial morphogenesis of the Drosophila follicle cell epithelium during oogenesis. Elimination of beta(H) by the karst mutation prevents apical constriction of the follicle cells during mid-oogenesis, and is accompanied by a gross breakup of the zonula adherens. We also report that the integrity of the migratory border cell cluster, a group of anterior follicle cells that delaminates from the follicle epithelium, is disrupted. Elimination of beta(H) prevents the stable recruitment of alpha-spectrin to the apical domain, but does not result in a loss of apicobasal polarity, as would be predicted from current models describing the role of spectrin in the establishment of cell polarity. These results demonstrate a direct role for apical (alphabeta(H))(2) spectrin in epithelial morphogenesis driven by apical contraction, and suggest that apical and basolateral spectrin do not play identical roles in the generation of apicobasal polarity. PMID- 10477759 TI - Filamin is required for ring canal assembly and actin organization during Drosophila oogenesis. AB - The remodeling of the actin cytoskeleton is essential for cell migration, cell division, and cell morphogenesis. Actin-binding proteins play a pivotal role in reorganizing the actin cytoskeleton in response to signals exchanged between cells. In consequence, actin-binding proteins are increasingly a focus of investigations into effectors of cell signaling and the coordination of cellular behaviors within developmental processes. One of the first actin-binding proteins identified was filamin, or actin-binding protein 280 (ABP280). Filamin is required for cell migration (Cunningham et al. 1992), and mutations in human alpha-filamin (FLN1; Fox et al. 1998) are responsible for impaired migration of cerebral neurons and give rise to periventricular heterotopia, a disorder that leads to epilepsy and vascular disorders, as well as embryonic lethality. We report the identification and characterization of a mutation in Drosophila filamin, the homologue of human alpha-filamin. During oogenesis, filamin is concentrated in the ring canal structures that fortify arrested cleavage furrows and establish cytoplasmic bridges between cells of the germline. The major structural features common to other filamins are conserved in Drosophila filamin. Mutations in Drosophila filamin disrupt actin filament organization and compromise membrane integrity during oocyte development, resulting in female sterility. The genetic and molecular characterization of Drosophila filamin provides the first genetic model system for the analysis of filamin function and regulation during development. PMID- 10477761 TI - Localized depolymerization of the major sperm protein cytoskeleton correlates with the forward movement of the cell body in the amoeboid movement of nematode sperm. AB - The major sperm protein (MSP)-based amoeboid motility of Ascaris suum sperm requires coordinated lamellipodial protrusion and cell body retraction. In these cells, protrusion and retraction are tightly coupled to the assembly and disassembly of the cytoskeleton at opposite ends of the lamellipodium. Although polymerization along the leading edge appears to drive protrusion, the behavior of sperm tethered to the substrate showed that an additional force is required to pull the cell body forward. To examine the mechanism of cell body movement, we used pH to uncouple cytoskeletal polymerization and depolymerization. In sperm treated with pH 6.75 buffer, protrusion of the leading edge slowed dramatically while both cytoskeletal disassembly at the base of the lamellipodium and cell body retraction continued. At pH 6.35, the cytoskeleton pulled away from the leading edge and receded through the lamellipodium as its disassembly at the cell body continued. The cytoskeleton disassembled rapidly and completely in cells treated at pH 5.5, but reformed when the cells were washed with physiological buffer. Cytoskeletal reassembly occurred at the lamellipodial margin and caused membrane protrusion, but the cell body did not move until the cytoskeleton was rebuilt and depolymerization resumed. These results indicate that cell body retraction is mediated by tension in the cytoskeleton, correlated with MSP depolymerization at the base of the lamellipodium. PMID- 10477762 TI - Regulated actin cytoskeleton assembly at filopodium tips controls their extension and retraction. AB - The extension and retraction of filopodia in response to extracellular cues is thought to be an important initial step that determines the direction of growth cone advance. We sought to understand how the dynamic behavior of the actin cytoskeleton is regulated to produce extension or retraction. By observing the movement of fiduciary marks on actin filaments in growth cones of a neuroblastoma cell line, we found that filopodium extension and retraction are governed by a balance between the rate of actin cytoskeleton assembly at the tip and retrograde flow. Both assembly and flow rate can vary with time in a single filopodium and between filopodia in a single growth cone. Regulation of assembly rate is the dominant factor in controlling filopodia behavior in our system. PMID- 10477763 TI - Regulation of cell contraction and membrane ruffling by distinct signals in migratory cells. AB - Cell migration and wound contraction requires assembly of actin into a functional myosin motor unit capable of generating force. However, cell migration also involves formation of actin-containing membrane ruffles. Evidence is provided that actin-myosin assembly and membrane ruffling are regulated by distinct signaling pathways in the migratory cell. Interaction of cells with extracellular matrix proteins or cytokines promote cell migration through activation of the MAP kinases ERK1 and ERK2 as well as the molecular coupling of the adaptor proteins p130CAS and c-CrkII. ERK signaling is independent of CAS/Crk coupling and regulates myosin light chain phosphorylation leading to actin-myosin assembly during cell migration and cell-mediated contraction of a collagen matrix. In contrast, membrane ruffling, but not cell contraction, requires Rac GTPase activity and the formation of a CAS/Crk complex that functions in the context of the Rac activating protein DOCK180. Thus, during cell migration ERK and CAS/Crk coupling operate as components of distinct signaling pathways that control actin assembly into myosin motors and membrane ruffles, respectively. PMID- 10477764 TI - Afadin: A key molecule essential for structural organization of cell-cell junctions of polarized epithelia during embryogenesis. AB - Afadin is an actin filament-binding protein that binds to nectin, an immunoglobulin-like cell adhesion molecule, and is colocalized with nectin at cadherin-based cell-cell adherens junctions (AJs). To explore the function of afadin in cell-cell adhesion during embryogenesis, we generated afadin(-/-) mice and embryonic stem cells. In wild-type mice at embryonic days 6.5-8.5, afadin was highly expressed in the embryonic ectoderm and the mesoderm, but hardly detected in the extraembryonic regions such as the visceral endoderm. Afadin(-/-) mice showed developmental defects at stages during and after gastrulation, including disorganization of the ectoderm, impaired migration of the mesoderm, and loss of somites and other structures derived from both the ectoderm and the mesoderm. Cystic embryoid bodies derived from afadin(-/-) embryonic stem cells showed normal organization of the endoderm but disorganization of the ectoderm. Cell cell AJs and tight junctions were improperly organized in the ectoderm of afadin( /-) mice and embryoid bodies. These results indicate that afadin is highly expressed in the ectoderm- derived cells during embryogenesis and plays a key role in proper organization of AJs and tight junctions of the highly expressing cells, which is essential for proper tissue morphogenesis. PMID- 10477765 TI - Distinct domains of MuSK mediate its abilities to induce and to associate with postsynaptic specializations. AB - Agrin released from motor nerve terminals activates a muscle-specific receptor tyrosine kinase (MuSK) in muscle cells to trigger formation of the skeletal neuromuscular junction. A key step in synaptogenesis is the aggregation of acetylcholine receptors (AChRs) in the postsynaptic membrane, a process that requires the AChR-associated protein, rapsyn. Here, we mapped domains on MuSK necessary for its interactions with agrin and rapsyn. Myotubes from MuSK(-/)- mutant mice form no AChR clusters in response to agrin, but agrin-responsiveness is restored by the introduction of rat MuSK or a Torpedo orthologue. Thus, MuSK( /)- myotubes provide an assay system for the structure-function analysis of MuSK. Using this system, we found that sequences in or near the first of four extracellular immunoglobulin-like domains in MuSK are required for agrin responsiveness, whereas sequences in or near the fourth immunoglobulin-like domain are required for interaction with rapsyn. Analysis of the cytoplasmic domain revealed that a recognition site for the phosphotyrosine binding domain containing proteins is essential for MuSK activity, whereas consensus binding sites for the PSD-95/Dlg/ZO-1-like domain-containing proteins and phosphatidylinositol-3-kinase are dispensable. Together, our results indicate that the ectodomain of MuSK mediates both agrin- dependent activation of a complex signal transduction pathway and agrin-independent association of the kinase with other postsynaptic components. These interactions allow MuSK not only to induce a multimolecular AChR-containing complex, but also to localize that complex to a primary scaffold in the postsynaptic membrane. PMID- 10477766 TI - Protein kinase C-dependent mobilization of the alpha6beta4 integrin from hemidesmosomes and its association with actin-rich cell protrusions drive the chemotactic migration of carcinoma cells. AB - We explored the hypothesis that the chemotactic migration of carcinoma cells that assemble hemidesmosomes involves the activation of a signaling pathway that releases the alpha6beta4 integrin from these stable adhesion complexes and promotes its association with F-actin in cell protrusions enabling it to function in migration. Squamous carcinoma-derived A431 cells were used because they express alpha6beta4 and migrate in response to EGF stimulation. Using function blocking antibodies, we show that the alpha6beta4 integrin participates in EGF stimulated chemotaxis and is required for lamellae formation on laminin-1. At concentrations of EGF that stimulate A431 chemotaxis ( approximately 1 ng/ml), the alpha6beta4 integrin is mobilized from hemidesmosomes as evidenced by indirect immunofluorescence microscopy using mAbs specific for this integrin and hemidesmosomal components and its loss from a cytokeratin fraction obtained by detergent extraction. EGF stimulation also increased the formation of lamellipodia and membrane ruffles that contained alpha6beta4 in association with F-actin. Importantly, we demonstrate that this mobilization of alpha6beta4 from hemidesmosomes and its redistribution to cell protrusions occurs by a mechanism that involves activation of protein kinase C-alpha and that it is associated with the phosphorylation of the beta4 integrin subunit on serine residues. Thus, the chemotactic migration of A431 cells on laminin-1 requires not only the formation of F-actin-rich cell protrusions that mediate alpha6beta4-dependent cell movement but also the disruption of alpha6beta4-containing hemidesmosomes by protein kinase C. PMID- 10477767 TI - CD38/ADP-ribosyl cyclase: A new role in the regulation of osteoclastic bone resorption. AB - The multifunctional ADP-ribosyl cyclase, CD38, catalyzes the cyclization of NAD(+) to cyclic ADP-ribose (cADPr). The latter gates Ca(2+) release through microsomal membrane-resident ryanodine receptors (RyRs). We first cloned and sequenced full-length CD38 cDNA from a rabbit osteoclast cDNA library. The predicted amino acid sequence displayed 59, 59, and 50% similarity, respectively, to the mouse, rat, and human CD38. In situ RT-PCR revealed intense cytoplasmic staining of osteoclasts, confirming CD38 mRNA expression. Both confocal microscopy and Western blotting confirmed the plasma membrane localization of the CD38 protein. The ADP-ribosyl cyclase activity of osteoclastic CD38 was next demonstrated by its ability to cyclize the NAD(+) surrogate, NGD(+), to its fluorescent derivative cGDP-ribose. We then examined the effects of CD38 on osteoclast function. CD38 activation by an agonist antibody (A10) in the presence of substrate (NAD(+)) triggered a cytosolic Ca(2+) signal. Both ryanodine receptor modulators, ryanodine, and caffeine, markedly attenuated this cytosolic Ca(2+) change. Furthermore, the anti-CD38 agonist antibody expectedly inhibited bone resorption in the pit assay and elevated interleukin-6 (IL-6) secretion. IL 6, in turn, enhanced CD38 mRNA expression. Taken together, the results provide compelling evidence for a new role for CD38/ADP-ribosyl cyclase in the control of bone resorption, most likely exerted via cADPr. PMID- 10477768 TI - Heterophilic binding of L1 on unmyelinated sensory axons mediates Schwann cell adhesion and is required for axonal survival. AB - This study investigated the function of the adhesion molecule L1 in unmyelinated fibers of the peripheral nervous system (PNS) by analysis of L1- deficient mice. We demonstrate that L1 is present on axons and Schwann cells of sensory unmyelinated fibers, but only on Schwann cells of sympathetic unmyelinated fibers. In L1-deficient sensory nerves, Schwann cells formed but failed to retain normal axonal ensheathment. L1-deficient mice had reduced sensory function and loss of unmyelinated axons, while sympathetic unmyelinated axons appeared normal. In nerve transplant studies, loss of axonal-L1, but not Schwann cell-L1, reproduced the L1-deficient phenotype. These data establish that heterophilic axonal-L1 interactions mediate adhesion between unmyelinated sensory axons and Schwann cells, stabilize the polarization of Schwann cell surface membranes, and mediate a trophic effect that assures axonal survival. PMID- 10477769 TI - The functional diversity of epidermal keratins revealed by the partial rescue of the keratin 14 null phenotype by keratin 16. AB - The type I epidermal keratins K14 and K16 are remarkably similar at the primary sequence level. While a structural function has been clearly defined for K14, we have proposed that a function of K16 may be to play a role in the process of keratinocyte activation that occurs after acute injury to stratified epithelia. To compare directly the functions of the two keratins we have targeted the expression of the human K16 cDNA to the progenitor basal layer of the epidermis of K14 null mice. Mice null for K14 blister extensively and die approximately 2 d after birth (Lloyd, C., Q.C. Yu, J. Cheng, K. Turksen, L. Degenstein, E. Hutton, and E. Fuchs. 1995. J. Cell Biol. 129:1329-1344). The skin of mice expressing K16 in the absence of K14 developed normally without evidence of blistering. However, as the mice aged they featured extensive alopecia, chronic epidermal ulcers in areas of frequent physical contact, and alterations in other stratified epithelia. Mice expressing a control K16-C14 cDNA also rescue the blistering phenotype of the K14 null mice with only a small percentage exhibiting minor alopecia. While K16 is capable of rescuing the blistering, phenotypic complementation in the resulting skin is incomplete due to the multiple age dependent anomalies. Despite their high sequence similarity, K16 and K14 are not functionally equivalent in the epidermis and other stratified epithelia and it is primarily the carboxy-terminal approximately 105 amino acids of K16 that define these differences. PMID- 10477775 TI - Recreational physical activity and the risk of cholecystectomy in women. AB - BACKGROUND: Physical activity may be an important determinant of the risk of gallstone disease in women, both independently and as a result of its role in maintaining body weight. METHODS: We prospectively studied recreational physical activity (such as jogging, running, and bicycling) and sedentary behavior (such as spending hours watching television) in relation to the risk of cholecystectomy, a surrogate for symptomatic cholelithiasis, in a cohort of 60,290 women who were 40 to 65 years of age in 1986 and had no history of gallstone disease. As part of the Nurses' Health Study, the women reported on questionnaires mailed to them every two years both their activity level and whether they had undergone cholecystectomy. During a 10-year follow-up period (1986 to 1996), 3257 cases of cholecystectomy were documented. RESULTS: Recreational physical activity was inversely related to the risk of cholecystectomy. The multivariate relative risk for women in the highest as compared with the lowest quintile of physical activity was 0.69 (95 percent confidence interval, 0.61 to 0.78). In contrast, sedentary behavior was independently related to an increased risk of cholecystectomy. As compared with women who spent less than 6 hours per week sitting while at work or driving, women who spent 41 to 60 hours per week sitting had a multivariate relative risk of 1.42 (95 percent confidence interval, 1.06 to 1.89), and women who spent more than 60 hours per week sitting while at work or driving had a multivariate relative risk of 2.32 (95 percent confidence interval, 1.26 to 4.26). These associations persisted after we controlled for body weight and weight change. CONCLUSIONS: In women, recreational physical activity is associated with a decreased risk of cholecystectomy. The association is independent of other risk factors for gallstone disease, such as obesity and recent weight loss. PMID- 10477776 TI - Increased mortality associated with growth hormone treatment in critically ill adults. AB - BACKGROUND: The administration of growth hormone can attenuate the catabolic response to injury, surgery, and sepsis. However, the effect of high doses of growth hormone on the length of stay in intensive care and in the hospital, the duration of mechanical ventilation, and the outcome in critically ill adults who are hospitalized for long periods is not known. METHODS: We carried out two prospective, multicenter, double-blind, randomized, placebo-controlled trials in parallel involving 247 Finnish patients and 285 patients in other European countries who had been in an intensive care unit for 5 to 7 days and who were expected to require intensive care for at least 10 days. The patients had had cardiac surgery, abdominal surgery, multiple trauma, or acute respiratory failure. The patients received either growth hormone (mean [+/-SD] daily dose, 0.10 +/- 0.02 mg per kilogram of body weight) or placebo until discharge from intensive care or for a maximum of 21 days. RESULTS: The in-hospital mortality rate was higher in the patients who received growth hormone than in those who did not (P<0.001 for both studies). In the Finnish study, the mortality rate was 39 percent in the growth hormone group, as compared with 20 percent in the placebo group. The respective rates in the multinational study were 44 percent and 18 percent. The relative risk of death for patients receiving growth hormone was 1.9 (95 percent confidence interval, 1.3 to 2.9) in the Finnish study and 2.4 (95 percent confidence interval, 1.6 to 3.5) in the multinational study. Among the survivors, the length of stay in intensive care and in the hospital and the duration of mechanical ventilation were prolonged in the growth hormone group. CONCLUSIONS: In patients with prolonged critical illness, high doses of growth hormone are associated with increased morbidity and mortality. PMID- 10477778 TI - The risk of recurrent deep venous thrombosis among heterozygous carriers of both factor V Leiden and the G20210A prothrombin mutation. AB - BACKGROUND: Point mutations in the factor V gene (factor V Leiden) and the prothrombin gene (the substitution of A for G at position 20210) are the most common causes of inherited thrombophilia. Whether or not factor V Leiden increases the risk of recurrent deep venous thrombosis is controversial, and there is no information on the risk of recurrence among carriers of both mutations. METHODS: We studied a retrospective cohort of 624 patients who were referred for a first episode of deep venous thrombosis. After excluding 212 patients with other inherited or acquired causes of thrombophilia, we compared 112 patients who were heterozygous carriers of factor V Leiden with 17 patients who were heterozygous for both factor V Leiden and the prothrombin mutation and 283 patients who had neither mutation. The relative risk of recurrent deep venous thrombosis was calculated with use of a proportional-hazards model. RESULTS: Patients who were heterozygous for factor V Leiden alone had a risk of recurrent deep venous thrombosis that was similar to that among patients who had neither mutation (relative risk, 1.1; 95 percent confidence interval, 0.7 to 1.6; P=0.76). In contrast, patients who were heterozygous for both factor V Leiden and the prothrombin mutation had a higher risk of recurrent thrombosis than did carriers of factor V Leiden alone (relative risk, 2.6; 95 percent confidence interval, 1.3 to 5.1; P=0.002). When the analysis was restricted to patients with spontaneous recurrences (i.e., ones that occurred in the absence of transient risk factors for venous thrombosis), the risk among carriers of both mutations, as compared with carriers of factor V Leiden alone, remained high (relative risk, 3.7; 95 percent confidence interval, 1.7 to 7.7; P<0.001), particularly if the first event had also been spontaneous (relative risk, 5.4; 95 percent confidence interval, 2.0 to 14.1; P<0.001). In contrast, the risk of recurrence in the presence of transient risk factors was similar among carriers of both mutations and carriers of factor V Leiden alone. CONCLUSIONS: The risk of recurrent deep venous thrombosis is similar among carriers of factor V Leiden and patients without this mutation. Carriers of both factor V Leiden and the G20210A prothrombin mutation have an increased risk of recurrent deep venous thrombosis after a first episode and are candidates for lifelong anticoagulation. PMID- 10477777 TI - A comparison of enoxaparin with placebo for the prevention of venous thromboembolism in acutely ill medical patients. Prophylaxis in Medical Patients with Enoxaparin Study Group. AB - BACKGROUND: The efficacy and safety of thromboprophylaxis in patients with acute medical illnesses who may be at risk for venous thromboembolism have not been determined in adequately designed trials. METHODS: In a double-blind study, we randomly assigned 1102 hospitalized patients older than 40 years to receive 40 mg of enoxaparin, 20 mg of enoxaparin, or placebo subcutaneously once daily for 6 to 14 days. Most patients were not in an intensive care unit. The primary outcome was venous thromboembolism between days 1 and 14, defined as deep-vein thrombosis detected by bilateral venography (or duplex ultrasonography) between days 6 and 14 (or earlier if clinically indicated) or documented pulmonary embolism. The duration of follow-up was three months. RESULTS: The primary outcome could be assessed in 866 patients. The incidence of venous thromboembolism was significantly lower in the group that received 40 mg of enoxaparin (5.5 percent [16 of 291 patients]) than in the group that received placebo (14.9 percent [43 of 288 patients]) (relative risk, 0.37; 97.6 percent confidence interval, 0.22 to 0.63; P< 0.001). The benefit observed with 40 mg of enoxaparin was maintained at three months. There was no significant difference in the incidence of venous thromboembolism between the group that received 20 mg of enoxaparin (43 of 287 patients [15.0 percent]) and the placebo group. The incidence of adverse effects did not differ significantly between the placebo group and either enoxaparin group. By day 110, 50 patients had died in the placebo group (13.9 percent), 51 had died in the 20-mg group (14.7 percent), and 41 had died in the 40-mg group (11.4 percent); the differences were not significant. CONCLUSIONS: Prophylactic treatment with 40 mg of enoxaparin subcutaneously per day safely and effectively reduces the risk of venous thromboembolism in patients with acute medical illnesses. PMID- 10477779 TI - Donor-derived acute promyelocytic leukemia in a liver-transplant recipient. PMID- 10477780 TI - Images in clinical medicine. Bacillus anthracis meningitis. PMID- 10477781 TI - Anthrax. PMID- 10477782 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 27-1999. An 82-year-old woman with numerous chronic disorders and a recent febrile illness. PMID- 10477783 TI - Exercise and the gallbladder. PMID- 10477785 TI - Correction: Effects of Different Forms of Dietary Hydrogenated Fats on Serum Lipoprotein Cholesterol Levels. PMID- 10477784 TI - Mission critical--integrating clinician-educators into academic medical centers. PMID- 10477786 TI - High levels of GM(1)-ganglioside and GM(1)-ganglioside beta-galactosidase in the parotid gland: a new model for secretory mechanisms of the parotid gland. AB - A new model for the subcellular basis of parotid secretion is presented in this article. GM(1)-ganglioside, typically found in neural tissues, is shown to be abundant in the parotid gland. This ganglioside may play a central role in membrane turnover mechanisms underlying exocytosis/endocytosis in its role as a promoter of membrane fusion or a fusogen. The lysosome and lysosomal hydrolases also play a central role in this model in catabolism of GM(1)-ganglioside. Consequently, high levels of the lysosomal hydrolase acidic beta-galactosidase are demonstrated in the salivary gland. GM(1)-gangliosidosis of the parotid glands, as described in mice, appears to be the first single-gene heritable disease found so far in the salivary glands. PMID- 10477787 TI - Acute viral and bacterial infections of the salivary glands. AB - Acute infection can involve any salivary gland, but it predominately affects the major salivary glands, especially the parotid gland. The anatomic and physiologic factors accounting for the parotid gland's predilection for infection are reviewed. Numerous conditions that are predisposed to acute bacterial sialadenitis and differ from risk factors associated with viral infection are also reviewed. The pathogenesis, diagnostic evaluation, treatment, complications, and prognosis of bacterial infections are discussed and contrasted with those of viral infections. PMID- 10477789 TI - Sialolithiasis. AB - Sialolithiasis is one of the most common problems that afflict the salivary glands and is a major cause of salivary gland dysfunction. Sialolithiasis is frequently encountered in clinical practice. The mechanism for stone formation is incompletely understood. The clinical diagnosis and standard management of sialolithiasis are discussed, and new modalities for treatment are also presented in this article. PMID- 10477788 TI - Chronic inflammatory disorders of the salivary glands. AB - When dealing with salivary gland disease, the otolaryngologist must be familiar with the various chronic inflammatory disorders that patients may be inflicted with. Physicians may have to treat such disorders as chronic sialadenitis, tuberculosis, sarcoidosis, animal scratch disease, actinomycosis, and Sjogren's syndrome. Symptoms, causes, and treatments of these chronic inflammatory disorders are discussed in this article. PMID- 10477790 TI - Noninflammatory, non-neoplastic disorders of the salivary glands. AB - Noninflammatory, non-neoplastic diseases comprise a diverse group of disorders that can present in a variety of ways. In general, a careful history will sharply focus the diagnostic possibilities. Although the diagnosis usually will be certain following the physical examination, additional testing may be needed to facilitate treatment planning. Sialolithiasis, cystic lesions, radiation injury, treatment to the parotid glands, sialodenosis, and other disorders are discussed in this article. PMID- 10477791 TI - Sjogren's syndrome. AB - Sjogren's syndrome is a rather common autoimmune disease primarily characterized by the dysfunction and destruction of exocrine glands associated with lymphocytic infiltrations. The disorder has a quite broad clinical presentation, ranging from glandular disease to systemic involvement and to the development of lymphoid malignancy. This article reviews the current aspects in clinical diagnosis and management and the immunopathogenesis of the disorder. PMID- 10477792 TI - Benign salivary gland neoplasms. AB - Benign salivary gland neoplasms represent a diverse group of neoplasms with varied clinical behaviors. Successful management of these tumors depends on accurate clinical and diagnostic assessment, followed by appropriate therapeutic intervention. An appropriate understanding of the biologic behavior of each tumor type allows for development of an appropriate treatment plan tailored to the individual patient. PMID- 10477793 TI - Malignant salivary gland neoplasms. AB - The treatment of salivary gland malignant neoplasms is challenging because of their infrequency, their unpredictable and varied biologic behavior, and their prolonged risk of recurrence. This article covers the pathophysiology of malignant neoplasms of the salivary glands. Emphasis is placed on the more common neoplasms. Histologic classification and management of neoplasms are also reviewed in this article. PMID- 10477794 TI - Surgery of the salivary glands. AB - Surgery of the salivary glands evolved rapidly after a greater understanding of the anatomy and pathologic processes affecting them was achieved. This article reviews the general and neurovascular anatomy of the major salivary glands. Understanding the anatomy of the salivary glands is a cornerstone of competent salivary gland surgery. The surgical techniques, potential pitfalls of surgery, and avoidance of postsurgical complications have been highlighted. Recent advances in surgical techniques are also presented. PMID- 10477795 TI - Trauma to the salivary glands. AB - Penetrating and blunt trauma to the salivary glands produces potential injury to the salivary tissue, salivary ducts, and to the facial nerve. This article presents a rationale of management for glandular and duct injury, emphasizing surgical and conservative therapy. It also presents management of neural injury including a discussion of surgical options and conservative management with electrical testing. PMID- 10477796 TI - The surgical pathology of salivary gland neoplasms. AB - The accurate and precise classification of salivary gland tumors is imperative. The various types of salivary gland tumors are distinct in their clinical behavior and response to treatment, and the surgical pathology diagnosis weighs heavily on therapeutic decisions and patient prognosis. The surgical pathology of salivary gland tumors is indeed challenging, but it need not be overwhelming. This review emphasizes those morphologic patterns and cellular features that are most helpful in accurately classifying the primary epithelial neoplasms most commonly encountered in the major and minor salivary glands. PMID- 10477797 TI - Management of facial nerve paralysis. AB - Facial paralysis is a dreaded complication of parotid surgery. It can lead to a variety of troubling symptoms in the patient, such as ocular problems and nasal obstruction. It can also have a significant emotional impact on the patient because of facial disfigurement and difficulties with communication, eating, and drinking. Successful outcome for patients inflicted with facial paralysis depends on proper selection of the appropriate rehabilitation technique. This article discusses the acute and late management of facial paralysis resulting from parotid surgery. PMID- 10477798 TI - Mechanisms of hepatic disposition of polystyrene microspheres in rats: effects of serum depend on the sizes of microspheres. AB - To study the mechanisms of the hepatic disposition of polystyrene microspheres (MS), effects of serum on their hepatic disposition characteristics were investigated for MSs with particle sizes of 50 nm (MS-50) and 500 nm (MS-500) by isolated liver perfusion experiments. It was revealed that serum in the perfusate inhibited and promoted the hepatic disposition of MS-50 and MS-500 at 37 degrees C, respectively. However, pre-heating at 56 degrees C or pre-treatment with anti C3 antibody of serum reduced the promotive effect of serum on the hepatic uptake of MS-500, suggesting that the complement system should be involved as opsonins for the hepatic uptake of MS-500. Hepatic disposition of both MSs at 4 degrees C was reduced by the addition of serum into the perfusate, which could be ascribed to the reduction of the surface hydrophobicity of MSs due to the adsorption of serum proteins onto the surface of MSs and to resultant decrease in non-specific disposition to the liver. From these results, serum was found to function both as the opsonin to enhance the hepatic uptake of MSs and as the inhibitor by reducing non-specific interaction between MSs and the plasma membrane. Whether serum promotes or inhibits the hepatic disposition of MSs would be dependent on the particle sizes of MSs. PMID- 10477799 TI - Release of a model molecule from highly concentrated fluorinated reverse emulsions. Influence of composition variables and temperature. AB - Highly concentrated reverse emulsions have been used to study the diffusion of a model molecule entrapped in these gel-emulsions. The influence of several parameters on the release of coumarin from fluorinated gel-emulsions has been investigated, and a computational method has been elaborated to determine the numerical value of the diffusion coefficients. The amount of probe molecule released depends on the initial loading amount, whereas the diffusion coefficient is not influenced by the initial concentration or by the amount of surfactant in the emulsions (in the range of the oil-to-surfactant ratios studied). The predominant factor seems to be the amount of water permitting the increase of the inter-phase area. Moreover, we have shown that the release of coumarin from gel emulsions is in accord with the 'Arrhenius' law and the 'activation energy' deduced can be due to a barrier counteracting the diffusion. PMID- 10477800 TI - Mechanisms controlling diffusion and release of model proteins through and from partially esterified hyaluronic acid membranes. AB - The effects of polymer percent esterification and protein molecular weight on the diffusion of two model proteins, deoxyribonuclease (DNase) and ribonuclease A (RNase A), through and from partially esterified hyaluronic acid membranes were compared. The permeability of the polymer membranes was inversely related to the degree of polymer esterification and the molecular weight of the protein. Transport rates of proteins through the membranes decreased dramatically over narrow ranges of polymer esterification. As expected, the apparent diffusivity of the larger protein in the polymer matrix was more sensitive to changes in membrane hydration than that of the smaller protein. These observations demonstrated the dependence of the mobility of large molecular weight proteins on polymer hydration and chain relaxation. The relationship between protein diffusion through and release from the modified hyaluronate matrices was also investigated using RNase A as a model. The release profiles from fully esterified membranes showed lag behavior and varied with protein load and hyaluronate hydrolysis rates, while release from less esterified membranes was rapid and independent of polymer esterification or hydrolysis. Potential applications of modified hyaluronate matrices in the controlled delivery of proteins are discussed. PMID- 10477801 TI - Drug delivery of anticancer agents: water soluble 4-poly (ethylene glycol) derivatives of the lignan, podophyllotoxin. AB - This paper reports on the synthesis and in vivo oncolytic activity of a series of water-soluble acyl derivatives of polyethylene glycol (PEG) conjugated podophyllotoxin. Some analogs of the polymer conjugate showed significantly better activity in a murine leukemia model than native podophyllotoxin suspended in an intralipid emulsion. Additionally, when tested intravenously against a solid lung tumor (A549) model, some conjugated analogs were equivalent to the podophyllotoxin/intralipid emulsion, while those compounds demonstrating slower rates of plasma hydrolysis (in vitro) appeared to cause greater toxicity. There appeared to be an overall correlation between the in vivo antitumor activity of the conjugate and its rate of hydrolysis in vitro, with those showing faster release possessing greater antitumor activity. In conclusion, the solubilization and predictable release of podophyllotoxin from a PEG carrier was achieved and resulted in some derivatives demonstrating, at a minimum, equivalency with podophyllotoxin when administered on an equal molar basis. Further studies may be warranted to assess the PEG-conjugates pharmacokinetics and therapeutic indices in leukemic models. PMID- 10477802 TI - Effect of polymer/basic drug interactions on the two-stage diffusion-controlled release from a poly(L-lactic acid) matrix. AB - We investigated the effect of drug physico-chemical properties on the release of basic drugs from poly(L-lactic acid) (P(L)LA) cylindrical matrices (rods; 10 mmx1 mm diameter). All the rods were revealed to exhibit two-stage diffusion controlled release profiles resulting from the transformation of P(L)LA from an amorphous to a semicrystalline state in aqueous medium. On the assumption that interactions between polymer carboxyl residues and basic drugs control the drug release rate, we evaluated the strength of these interactions by the drug partition between the polymer and the aqueous medium. In the first release stage, the drugs diffused through the swollen polymer matrix. The polymer-drug interactions shielded the polymer terminal carboxyl residues, thereby resulting in a less hydrated matrix and consequent diminishment of drug diffusion. In the second release stage, the drugs diffused through the water-filled micropores which had developed as a result of polymer crystallization. The stronger polymer basic drug interactions reduced the drug diffusion rate by decreasing not only the porosity of the matrix, but also the drug partition to the water-filled micropores. It was also found that the fractional drug release rate in the second stage increased with drug content of the rod at the pH where both the polymer carboxyl residues and the drugs were ionized. Since the polymer-drug interactions must be close to saturation with increasing drug content, we believe this result to be due to an increase in the ratio of the drug partition to the water-filled micropores. PMID- 10477803 TI - Poly(D,L-lactic-co-glycolic acid) microspheres for sustained delivery and stabilization of camptothecin. AB - Camptothecin (CPT) and its water-insoluble derivatives are known as topoisomerase I inhibitors exhibiting high antitumoral activity against a wide spectrum of human malignancies. Until now clinical application of CPT is restricted by insolubility and instability of the drug in its active lactone form resulting in less antitumor potency and poor bioavailability. For these reasons CPT-loaded microspheres were prepared by the solvent evaporation method using the H-series of poly(D,L-lactide-co-glycolide) (H-PLGA), which contain more carboxylic acid end chains and hydrate faster than the non-H-series. At 1.2% CPT-payload the drug was molecular dispersed throughout the matrix whereas at higher CPT-payload the amount of crystalline CPT-islets increased with the CPT content. The release pattern of CPT was biphasic comprising a first burst effect delivering 20-35% of the payload and increasing with drug-loading. This phase was followed by sustained delivery of CPT releasing 40-75% of the payload within 160 h. In comparison to PLGA-microspheres, the CPT-release rate from H-PLGA was twofold higher and accelerated. The active CPT-lactone was maintained during preparation, storage and release due to hindered diffusion of acidic oligomers among other mechanisms. Thus stabilization and sustained release of CPT from PLGA microspheres might reduce local toxicity combined with prolonged efficacy offering new perspectives in CPT chemotherapy. PMID- 10477804 TI - Design of an intravaginal ring for the controlled delivery of 17 beta-estradiol as its 3-acetate ester. AB - Suitable ester prodrugs of 17beta-estradiol are identified, thus permitting effective sustained and controlled estrogen replacement therapy (ERT) from an elastomeric, silicone intravaginal ring (IVR). IVR devices of reservoir design were prepared by blending silicone elastomer base with n-propylorthosilicate (cross-linker) and 10% w/w of 17beta-estradiol or an ester prodrug, the mix being activated with 0.5% w/w stannous octoate and cured at 80 degrees C for 2 min. A rate-controlling membrane was similarly prepared, without the active agent. IVR devices were of cross-sectional diameter 9 mm, outer diameter 54 mm, with core cross-sectional diameter of 2 mm and core length varied as required. Sink conditions were evident for the 17beta-estradiol esters in 1.0% aqueous benzalkonium chloride solution. The low release rates into 0.9% w/v saline of the lipophilic valerate and benzoate esters were due to their intrinsically low aqueous solubilities. In vivo, these esters failed to raise plasma estradiol above baseline levels in postmenopausal human volunteers, despite good in vitro release characteristics under sink conditions. The best release rates under sink conditions, in combination with substantial aqueous solubilities as indicated by the release rates into saline, were observed for the acetate and propionate esters. A combination of drug release characteristics, short plasma half-life and a toxicologically acceptable hydrolysis product indicated that 17beta-estradiol-3 acetate was the prodrug of choice for IVR delivery of ERT. In vivo, an IVR device releasing 100 microg/day of estradiol as its 3-acetate ester maintained over 84 days a circulating plasma concentration in the region of 300 pmoll(-1), within the clinically desirable range for ERT. PMID- 10477805 TI - Uptake and release of budesonide from mucoadhesive, pH-sensitive copolymers and their application to nasal delivery. AB - Microparticles of novel, bioadhesive graft copolymers of polymethacrylic acid and polyethylene glycol (P(MAA-g-EG)) were prepared. The aims of this study were to investigate the uptake and release kinetics of budesonide from P(MAA-g-EG) in vitro as well as the pharmacokinetics following nasal administration of the polymer contained budesonide. The loading of budesonide into the pH-sensitive polymers was examined using various ethanol solutions. Ethanol was required for drug solubilization but hindered hydrogel swelling at pH 7.2. Maximum loading of the drug in the polymer was obtained using 25% ethanol solutions. The release of budesonide from the polymer swollen in 25% ethanol solutions obeyed classical Fickian release behavior after an initial rapid drug burst. For nasal administration of budesonide-containing P(MAA-g-EG) the plasma concentration of budesonide was kept constant following a peak concentration of the drug approximately 45 min after administration. PMID- 10477806 TI - Local delivery of chlorhexidine using a tooth-bonded delivery system. AB - Films containing 20% w/w chlorhexidine base (particle size 63-125 microm) in poly(epsilon-caprolactone), MW 35,000-45,000, were prepared by solvent evaporation and sections attached to the mesio-lingual and mesio-buccal surfaces of the lower first molar in healthy volunteers. Saliva (<1.5 microl) was collected on Periopaper and chlorhexidine concentrations measured by HPLC were typically higher in the area immediately adjacent to the tooth-bonded film sections and lower at more distant sites. Analysis of variance of chlorhexidine concentrations, adjacent to the film sections, showed concentrations were significantly different on the buccal and lingual sides of the tooth and depended on the time of sampling (n=5, P<0.05). PMID- 10477807 TI - A mathematical model of an aqueous-organic partition-based controlled release system using microporous membranes. AB - A mathematical model with an exact solution is presented for the membrane controlled release of small molecules such as nicotine, caffeine, and benzoic acid initially present in solution in the reservoir of the device. Both hollow fiber and flat membrane device geometries are considered. The reservoir is bounded by a microporous membrane, the pores of which are filled with a pore liquid immiscible with the reservoir phase liquid. At the interface between the reservoir and the pore, the solute partitions between the reservoir and the pore liquid phases, before diffusing outward through the membrane pore. The model results compare well with experimental data. Parametric studies reveal the interaction between system parameters and the controlled release behavior. A high partition coefficient of the solute between the reservoir and pore phases is found to effect pseudo-zero order release for an extended time. Similarly, when the ratio of time constants for transport of the solute through the reservoir and membrane regions is small, a constant release rate is achieved for an extended time. PMID- 10477809 TI - Effect of compression speeds on the compaction properties of a 1:1 paracetamol microcrystalline cellulose mixture prepared by single compression and by combinations of pre-compression and main-compression. AB - A 1:1 blend of paracetamol and microcrystalline cellulose was compacted at different compression speeds by single compression or combinations of pre- and main-compression. The tensile strengths of the tablets decreased from 0.74+/-0.01 to 0.44+/-0.05 MPa as the compression speed was increased from 78 to 390 mm/s when a single compression pressure of 80 MPa was used to compress the tablets. When combinations of pre- and main-compression of 320 and 240 MPa were used to compress the tablets, tensile strengths decreased from 3.12+/-0.67 MPa at a compression speed of 78 mm/s to 1.24+/-0.36 MPa when the compression speed was 390 mm/s. The energies of compression and the ratio of elastic to plastic energies increased with increase in compression speed. This was because the material was becoming more elastic and more energy was required for the elastic expansion leading to a reduction in the energy available for plastic deformation and bond formation which resulted in a decrease in tensile strengths. Pre compression played a major role at high compression speeds. The tensile strengths of tablets (1.2+/-0.08 MPa) compressed with a pre-compression of 160 MPa followed by a main-compression of 80 MPa (compression speed of 390 mm/s) were similar to the tensile strengths of tablets (1.1+/-0.10 MPa) compressed using a single compression of 320 MPa at the same compression speed of 390 mm/s. Thus, combinations of lower pressures can be employed to compress the material to the same tensile strength as a high single compression. PMID- 10477808 TI - Microencapsulation of DNA using poly(DL-lactide-co-glycolide): stability issues and release characteristics. AB - The design of DNA vaccination delivery systems for the targeting of professional antigen presenting cells could be an interesting approach to elicit cytotoxic T cell responses to fight viral infections and in cancer therapy. Stability studies with linear high and low molecular DNA and supercoiled plasmid DNA were performed in order to check their ability to withstand stress conditions applied during formulation processes. DNA was tested for integrity by the PicoGreen assay and transfectivity was assessed in cell culture transfection experiments. Double stranded DNA is extremely stable under physiological conditions in vitro but is rapidly degraded under acidic conditions and high shear forces. Thereby, different stress factors resulted in distinct degradation patterns such as fragmentation and strand separation possibly followed by further decomposition of single-stranded DNA. DNA containing PLGA microparticles as a potential delivery system was prepared by spray-drying. Encapsulation efficiency, DNA stability and burst release varied significantly depending on the different parameters explored in this study. The microencapsulation process was altered to achieve maximal stability of encapsulated DNA by reducing exposure to shear forces and by the addition of NaHCO(3) which acts as a buffering agent and furthermore stabilizes dsDNA against mechanical degradation. Stability of DNA is maintained during the burst release phase, but massive degradation occurred during the second release phase possibly due to acidic catalyzed decomposition. In summary, we feel that microencapsulation of DNA vaccines by spray-drying offers manifold possibilities to design suitable delivery systems in terms of optimizing phagocytosis by APCs and maintaining stability of DNA in phagosomes. PMID- 10477810 TI - Determination of ornidazole in pharmaceutical dosage forms based on reduction at an activated glassy carbon electrode. AB - The electrochemical reduction of ornidazole was studied at a glassy carbon electrode activated by applying a new pretreatment. The dependence of intensities of currents and potentials on pH, concentration, scan rate, nature of the solvent (aqueous media, mixed aqueous-organic systems) and surfactant was investigated. Linear calibration plots were obtained over the concentration ranges 4x10(-6) 6x10(-4) and 6x10(-6)-6x10(-4) mol l(-1) in 0.2 M H(2)SO(4) and acetate buffer (pH 4.7), respectively. The method was applied to the determination of ornidazole in different drug formulations. PMID- 10477811 TI - Beclomethasone dipropionate 250 ug per dose metered dose inhalers: effect of Volumatic spacer on potentially respirable dose. AB - The aerodynamic particle size distribution of beclomethasone dipropionate from 250 ug per dose metered dose inhalers tested in conjunction with a Volumatic spacer, was studied using a multistage liquid impinger. The products tested were Becloforte (Allen and Hanburys), Becloforte (parallel import from Spain), Beclazone 250 (Norton Healthcare) and Filair Forte (3M Health Care). Tests were performed with either zero or 3 s time delay between actuation of the inhaler and sampling. Filair Forte gave significantly lower deposition than all the other products in both the <6.8 and <3.1 um size ranges with both zero and 3 s residence times (49.8% less than Becloforte, UK-origin, at <6.8 um, zero residence time). This was despite fitting well in the Volumatic. Testing a second batch of Filair Forte confirmed the low deposition results. The only other significant differences were greater depositions from Becloforte (Spanish) compared to Beclazone 250 (<6.8 um, zero residence time and <3.1 um, 3 s residence time) and between Becloforte (Spanish) and all the other products (<3.1 um, zero residence time). The magnitude of these differences were, however, less marked than those shown by Filair Forte. Increasing the residence time of the aerosol in the Volumatic to 3 s, simulating how patients may use the inhaler/spacer combination, gave similar results to those obtained with zero residence time. Filair Forte, used with the Volumatic, delivered a relatively low dose in the potentially respirable size range, compared to Becloforte. No substantial differences were seen at <6.8 um between Becloforte (UK or Spanish) or Beclazone 250. Further work would be required to determine the importance of the differences observed between the products in the <3.1 um range. PMID- 10477812 TI - Cefpodoxime-proxetil hydrolysis and food effects in the intestinal lumen before absorption: in vitro comparison of rabbit and human material. AB - The luminal and mucosal deesterification of the prodrug ester cefpodoxime proxetil was studied in human duodenal washings in vitro. Enzymatic hydrolysis of the ester, releasing the active third generation cephalosporin, was observed in luminal washing in the same way as it had previously been observed in the rabbit. Eserine and PMSF and HgCl(2) were potent inhibitors of cefpodoxime-proxetil hydrolysis in luminal washing, suggesting the participation of a cholinesterase in the hydrolysis of cefpodoxime-proxetil. These results are in agreement with our previous findings performed in the rabbit. Moreover, cefpodoxime-proxetil directly decreases the acetylcholinesterase activity when tested by a specific enzymatic method. These observations support the hypothesis that the partial oral bioavailability of cefpodoxime-proxetil results from hydrolysis by luminal cholinesterases. In vitro experiments run with rabbit duodenal washing with food components were compared with the pharmacokinetics of cefpodoxime-proxetil in humans. Amino acids, trace elements and vitamins were potent inhibitors for cefpodoxime-proxetil hydrolysis in duodenal washings. Otherwise, lipids (LTC and mixed LCT/MCT) did not interact. In the human, cefpodoxime-proxetil bioavailability is significantly enhanced when tablets are administered with food. The correlation found between animal results and human results in vitro for prospective investigation of a new prodrug ester could be very useful. An in vitro hydrolysis in intestinal animal washings could allow the potentially degraded condition and the food effect of the luminal tract to be assessed before absorption. PMID- 10477813 TI - Rotating dialysis cell as in vitro release method for oily parenteral depot solutions. AB - The purpose of the study was to investigate an in vitro release method based on a rotating dialysis cell for parenteral oil depot formulations using different model conditions and test formulations. The total amount of drug released from the rotating dialysis cell was in accordance with the theoretical values calculated from the partition coefficients. The release rates were shown to depend on the total amount of drug available for the release process and to follow first order kinetics. The rotating dialysis cell has a potential as in vitro release method for characterization of oily depot formulations for parenteral administration. PMID- 10477814 TI - Freeze-drying of drug-free and drug-loaded solid lipid nanoparticles (SLN). AB - Solid lipid nanoparticles (SLN) of a quality acceptable for i.v. administration were freeze-dried. Dynasan 112 and Compritol ATO 888 were used as lipid matrices for the SLN, stabilisers were Lipoid S 75 and poloxamer 188, respectively. To study the protective effect of various types and concentrations of cryoprotectants (e.g. carbohydrates), freeze-thaw cycles were carried out as a pre-test. The sugar trehalose proved to be most effective in preventing particle growth during freezing and thawing and also in the freeze-drying process. Changes in particle size distribution during lyophilisation could be minimised by optimising the parameters of the lyophilisation process, i.e. freezing velocity and redispersion method. Lyophilised drug-free SLN could be reconstituted in a quality considered suitable for i.v. injection with regard to the size distribution. Loading with model drugs (tetracaine, etomidate) impairs the quality of reconstituted SLN. However, the lyophilisate quality is sufficient for formulations less critical to limited particle growth, e.g. freeze-dried SLN for oral administration. PMID- 10477815 TI - A new generation starch product as excipient in pharmaceutical tablets. III. Parameters affecting controlled drug release from tablets based on high surface area retrograded pregelatinized potato starch. AB - This paper describes the general applicability of a new pregelatinized starch product in directly compressible controlled-release matrix systems. It was prepared by enzymatic degradation of potato starch followed by precipitation (retrogradation), filtration and washing with ethanol. The advantages of the material include ease of tablet preparation, the potential of a constant release rate (zero-order) for an extended period of time and the possibility to incorporate high percentages of drugs with different physicochemical properties. Constant release profiles are the result of solvent penetration into the tablet. For theophylline as test drug, constant release profiles could be realized up to a drug content of 75%. This illustrates the possibility to control the release of highly dosed drugs. Release rates from retrograded pregelatinized starch tablets can be enhanced or decreased to the desired profile by different parameters, like geometries of the tablet, compaction force and the incorporation of additional excipients. For procaine HCl it is demonstrated that larger tablets show slower release rates. The incorporation of soluble excipients like lactose and mannitol results for paracetamol in enhanced release rates. The delivery of bases and their salts can be modified by the incorporation of organic acid or alkaline excipients, as is demonstrated for lidocaine and procaine HCl. PMID- 10477816 TI - Preparation, characterization and cytotoxicity of methylmethacrylate copolymer nanoparticles with a permanent positive surface charge. AB - Methylmethacrylate copolymer nanoparticles containing different cationic comonomers such as N-trimethylammoniumethylmethacrylate (TMAEMC), N dimethylammoniumethylmethacrylate (DMAEMC), N trimethylammoniumpropylmethylacrylamide (MAPTAC) or the anionic comonomer sulfopropylmethacrylate (SPM), respectively, were prepared by free radical polymerization. Particle size was determined by photon correlation spectroscopy (PCS), transmission and scanning electron microscopy (TEM, SEM), and surface charge by microelectrophoresis. Pure poly(methylmethacrylate) nanoparticles served as control. Depending on the method, mean diameters of permanently positively-charged nanoparticles MMA-TMAEMC and MMA-MAPTAC were 243 or 207 nm (PCS), 161 or 201 nm (TEM), and 158 or 197 nm (SEM), respectively. Zeta potential examined in demineralized water or NaCl solution was +63.4 or +32.1 mV for MMA TMAEMC nanoparticles and +49.2 or +32.0 mV for MMA-MAPTAC nanoparticles, respectively. Cytotoxicity of nanoparticles was determined by MTT assay in three different cell cultures including human foreskin fibroblasts (HFF) and two monkey kidney cell lines MA-104 and Vero. Cell viability profiles of TMAEMC and MAPTAC containing nanoparticles were different, showing IC(50) values for MMA-TMAEMC nanoparticles of 189.6+/-11.4 ug/ml (MA-104), 110.9+/-3.1 ug/ml (Vero) and 27.2+/ 4.0 ug/ml (HFF). Cell viability at maximum concentration of 500 ug/ml MMA-MAPTAC nanoparticles was 98.3% (Vero), 85.7% (MA-104), or 94.0% (HFF), respectively. PMID- 10477817 TI - Diffusion of glucose and insulin in a swelling N-isopropylacrylamide gel. AB - The diffusional characteristics for poly(N-isopropylacrylamide) (NiPAAm) gel have been investigated. This gel is a critical gel which means that small changes in the environment influence the gel volume considerably. The effective diffusion coefficients for the solutes glucose and insulin were determined in batch experiments with the solutes diffusing out from small cylindrical gel bodies with diameters of 2.4-2.9 mm and at temperatures below the critical temperature: 10, 20 and 30 degrees C. The effective diffusion coefficients were obtained by fitting the experimental data to a mathematical model considering back-mixing and time delay in the experimental set-up, dilution due to the adsorbed liquid on the gel bodies and partition due to the exclusion effect. The effective diffusion coefficient for glucose increases from 2.7.10(-10) to 4.7.10(-10) m(2)/s when the temperature increases from 10 to 30 degrees C, following the Wilke-Chang relationship. This implies that the effect of the network is negligible compared with the effect of the temperature. However, for a solute with a molecular weight of about 6000 the network has a considerable effect. The effective diffusion coefficient for insulin increases from 4.4.10(-10) to 5.9.10(-10) m(2)/s when the temperature increases from 10 to 30 degrees C, which is less than expected from the Wilke-Chang relationship. This indicates an increased resistance for diffusion inside the gel due to shrinking. PMID- 10477818 TI - Applications of pressure differential scanning calorimetry in the study of pharmaceutical hydrates. I. Carbamazepine dihydrate. AB - The dehydration of carbamazepine dihydrate (C(15)H(12)N(2)O.2H(2)O) was studied by both conventional differential scanning calorimetry (DSC) and by pressure differential scanning calorimetry. Variable temperature powder X-ray diffractometry (VTXRD) and thermogravimetric analysis were used as complementary techniques. By performing DSC at elevated pressures, the dehydration and vaporization endotherms were separated and it was possible to determine the enthalpy of dehydration. Over the range of 100-600 psi, the enthalpy of dehydration was unaffected by pressure. However, the solid-state of the anhydrous phase formed was influenced by the DSC conditions. At ambient pressure, dehydration resulted in the formation of the gamma-form of anhydrous carbamazepine while at elevated pressures, the anhydrous beta-form appeared which converted to the gamma-form at higher temperatures. At high pressures, the water liberated on dehydration is not immediately removed and its presence appears to be responsible for the formation of beta-carbamazepine. VTXRD permitted in situ study of dehydration and this confirmed the DSC results. A specially fabricated sample holder permitted VTXRD studies at elevated pressures. PMID- 10477819 TI - The mobile order solubility equation applied to polyfunctional molecules: The non hydroxysteroids in aqueous and non aqueous solvents. AB - The solubility equation for real solutions derived from the thermodynamics of mobile order in liquids is used to predict the solubility of non-hydroxysteroids in water and in common polar and nonpolar organic solvents. Strictly obtained on a thermodynamic basis, the model allows not only correct predictions of the solubilities from the knowledge of a limited number of characteristics of solutes and solvents, but also enables a better understanding of the solution process and of the factors that determine solubility. Some practical rules are derived which might help to orient the choice of a solvent for liquid pharmaceutical forms. PMID- 10477820 TI - Stabilization of gene delivery systems by freeze-drying. AB - Freeze-drying of three different forms of gene delivery systems was performed using a controlled two-step drying process and 10% sucrose as lyoprotectant. Complexes of pCMVL plasmid with transferrin-conjugated polyethylenimine, adenovirus-enhanced transferrinfection consisting of pCMVL/transferrin-polylysine complexes linked to inactivated adenovirus particles, and a recombinant, E1 defective adenovirus expressing a luciferase reporter gene were tested. Three weeks after freeze-drying the reagents were rehydrated with water and tested for transfection activity. Luciferase gene expression levels were retained at high levels in all three systems, in contrast to reagents stored in solution. The use of the lyoprotectant was essential. In the absence of sucrose the transfection activities dropped by a factor of 100-1000. The data suggest freeze-drying as a useful method for stabilization and storage of standardized batches of transfection agents. PMID- 10477821 TI - Study of the complexation behaviour of gliclazide with partially methylated beta cyclodextrin in solution and solid state. AB - The complexation of Gliclazide (GL) with a partially methylated beta-cyclodextrin was studied. Phase-solubility and (1)H NMR spectroscopy were employed to investigate the complexation behaviour in solution and to demonstrate the complexation in liquid medium with the participation of both azabicyclooctyl and tolyl moieties of GL in the inclusion process. Solid systems prepared by kneading, co-grinding and spray drying have also been checked, using DSC and HSM, for assessing the formation of the inclusion compound. Experimental evidence of the complexation between drug and cyclodextrin was reported for the co-ground and spray-dried systems. PMID- 10477822 TI - Cell and molecular biology of the assembly and secretion of apolipoprotein B containing lipoproteins by the liver. AB - Triglycerides are one of the most efficient storage forms of free energy. Because of their insolubility in biological fluids, their transport between cells and tissues requires that they be assembled into lipoprotein particles. Genetic disruption of the lipoprotein assembly/secretion pathway leads to several human disorders associated with malnutrition and developmental abnormalities. In contrast, patients displaying inappropriately high rates of lipoprotein production display increased risk for the development of atherosclerotic cardiovascular disease. Insights provided by diverse experimental approaches describe an elegant biological adaptation of basic chemical interactions required to overcome the thermodynamic dilemma of producing a stable emulsion vehicle for the transport and tissue targeting of triglycerides. The mammalian lipoprotein assembly/secretion pathway shows an absolute requirement for: (1) the unique amphipathic protein: apolipoprotein B, in a form that is sufficiently large to assemble a lipoprotein particle containing a neutral lipid core; and, (2) a lipid transfer protein (microsomal triglyceride transfer protein-MTP). In the endoplasmic reticulum apolipoprotein B has two distinct metabolic fates: (1) entrance into the lipoprotein assembly pathway within the lumen of the endoplasmic reticulum; or, (2) degradation in the cytoplasm by the ubiquitin dependent proteasome. The destiny of apolipoprotein B is determined by the relative availability of individual lipids and level of expression of MTP. The dynamically varied expression of cholesterol-7alpha-hydroxylase indirectly influences the rate of lipid biosynthesis and the assembly and secretion lipoprotein particles by the liver. PMID- 10477823 TI - Lateral organisation of membrane lipids. The superlattice view. AB - Most biological membranes are extremely complex structures consisting of hundreds or even thousands of different lipid and protein molecules. The prevailing view regarding the organisation of these membranes is based on the fluid-mosaic model proposed by Singer and Nicholson in 1972. According to this model, phospholipids together with some other lipids form a fluid bilayer in which these lipids are diffusing very rapidly laterally. The idea of rapid lateral diffusion implies that, in general, the different lipid species would be randomly distributed in the plain of the membrane. However, there are recent data indicating that the components tend to adopt regular (superlattice-like) distributions in fluid, mixed bilayers. Based on this, a superlattice model of membranes has been proposed. This superlattice model is intriguing because it allows only a limited certain number of 'critical' compositions. These critical compositions could play a key role in the regulation of the lipid compositions of biological membranes. Furthermore, such putative critical compositions could explain how compositionally distinct organelles can exist despite of rapid inter-organelle membrane traffic. In this review, these intriguing predictions are discussed along with the basic principles of the model and the evidence supporting it. PMID- 10477824 TI - Dissociation of surfactant protein B from canine surfactant large aggregates during formation of small surfactant aggregates by in vitro surface area cycling. AB - Pulmonary surfactant isolated by lavage can be separated into large aggregates (LA) and small aggregates (SA). Pulse labeling experiments have shown that the LA subtype is the precursor of the SA subtype. Conversion of LA to SA can be demonstrated in vitro using the technique of surface area cycling. The precise mechanisms of surfactant subtype conversion remain unknown. We have previously reported a decline in surfactant-associated protein B (SP-B) during in vitro subtype conversion of canine surfactant. This led to the hypothesis that SP-B may be degraded by a serine protease 'convertase' during cycling. The current studies used a quantitative slot-blot assay to investigate the fates of SP-A and SP-B during in vitro cycling. These studies confirmed some SP-A is present in SA, but SP-B is confirmed to LA. Conversion leads to an apparent loss of SP-B during cycling. However, SP-B can be recovered from the walls of polypropylene and Teflon tubes by washing with chloroform:methanol. Recovered SP-B migrated on non reducing tricine gels as a single band with an apparent molecular weight of 17 kDa, corresponding to intact SP-B dimer. Reconstitution studies demonstrated that the recovered SP-B retained its surface active properties as determined on a pulsating bubble surfactometer. We conclude in vitro surface area cycling of canine LA results in the dissociation of SP-B from surfactant lipids resulting in an apparent decline in SP-B levels. PMID- 10477825 TI - Enzymatic properties of rat group IIA and V phospholipases A(2) compared. AB - Group IIA and V phospholipases A(2) (PLA(2)s) are known to play a role in inflammatory responses. We have constructed a bacterial expression vector for rat group IIA and V PLA(2)s, over-expressed, folded and purified the proteins with the aim to study and compare the properties of the enzymes in detail. For zwitterionic phospholipid micelles, both enzymes display optimum activity at pH 8. 0 and absolutely require Ca(2+) for enzymatic activity. In the presence of substrate, group V PLA(2) has a high affinity for Ca(2+) (K(Ca2+)=90 microM) while K(Ca2+) of group IIA PLA(2) was found to be 1.6 mM. The absence of substrate only marginally influences the Ca(2+) affinities. In contrast to group IIA PLA(2), group V PLA(2) does not show a jump in the activity profile at substrate concentrations around the critical micelle concentration. Direct binding studies using n-alkylphosphocholines indicate that group V PLA(2) forms protein-lipid aggregates at pre-micellar lipid concentrations in a cooperative and Ca(2+)-dependent manner. This behavior, which is comparable to that observed for the PLA(2) from Naja melanoleuca snake venom, reflects the high affinity of this enzyme for zwitterionic phospholipids. Competitive inhibition by the substrate analogues (R)-2-dodecanoylaminohexanol-1-phosphocholine and its phosphoglycol derivative was tested on zwitterionic micelles as substrate. Group IIA PLA(2) shows a preference for the phosphoglycol inhibitor whereas the phosphocholine inhibitor binds stronger to the active site of group V PLA(2). The enzymatic activity was also measured on zwitterionic liposomes which appear to be much better substrates for group V PLA(2) than for group IIA PLA(2). The overall results suggest that group V PLA(2) is better suited for action on biological membranes than group IIA PLA(2). PMID- 10477826 TI - Transfer and subsequent metabolism of lysolipids studied by immobilizing subcellular compartments in alginate beads. AB - The transfer and subsequent metabolism of lysophosphatidylcholine between subcellular compartments were studied in vitro by embedding membranes in alginate beads. After several experiments to validate the process, it was demonstrated that lysophosphatidylcholine was transferred from microsomes embedded in alginate beads to immobilized chloroplasts, that this transfer involved the partition of this molecule, and that the imported lysophosphatidylcholine was further used as substrate for phosphatidylcholine biosynthesis. More generally, the technique used makes it possible to avoid any cross-contamination between compartments, to evidence a transfer of molecules, and to study the metabolism of the imported molecules in the acceptor compartment. PMID- 10477827 TI - Up-regulation of uterine UCP2 and UCP3 in pregnant rats. AB - Pregnancy produces profound changes in hormone dynamics, thermoregulation and energy metabolism. Uncoupling proteins (UCPs) have been identified in a variety of tissues and UCP1 is known to play important roles in energy homeostasis, while the regulation of UCP2 and UCP3 is still unclear. The present study aimed to investigate the effects of the changes during pregnancy on UCP gene expression in the uterus, as well as in brown adipose tissue (BAT), white adipose tissue (WAT), soleus muscle (Muscle), and liver, throughout the estrus and metestrus periods, at early, middle and late stages in pregnancy, and during post-gestational stages. The expression of uterine UCP2 and UCP3 were up-regulated by 3.2- and 1. 5-fold, respectively, during the late stage of pregnancy with an increase of WAT leptin mRNA expression and exogenous administration of leptin resulted in induction of the uterine UCP2 and UCP3 levels. Contrary to uterine UCPs, UCP1 mRNA expression in BAT was down-regulated by 0.5-fold and there were no remarkable changes in WAT or liver UCP2, or Muscle UCP3 expression throughout the periods. These results indicate that UCP gene expressions during pregnancy are regulated tissue-dependently, and up-regulation of uterine UCP2 and UCP3 mRNA may be due to increased leptin levels. PMID- 10477828 TI - Phosphatidic acid increases inositol-1,4,5,-trisphosphate and [Ca2+]i levels in neonatal rat cardiomyocytes. AB - Phosphatidic acid (PA), which can be synthesized de novo, or as a product of phosphatidylcholine hydrolysis and/or phosphorylation of 1,2-diacylglycerol (DAG), mediates diverse cellular functions in various cell types, including cardiomyocytes. We set out to characterize the effect of PA on intracellular free calcium ([Ca2+]i) and inositol-1,4,5-trisphosphate (IP(3)) levels in primary cultures of neonatal rat cardiomyocytes. Addition of PA led to rapid, concentration and time dependent increases in both IP(3) and [Ca2+]i levels in adherent cells. There was strong correlation in the concentration-response relationships between IP(3) and [Ca2+]i increases evoked by PA. Incubation with the sarcoplasmic reticulum (SR) Ca2+ pump inhibitor, cyclopiazonic acid (CPA), significantly attenuated the PA evoked [Ca2+]i increase but had no significant effect on IP(3) accumulation. The phospholipase C (PLC) inhibitor, D-609, attenuated both IP(3) and [Ca2+]i elevations evoked by PA whereas staurosporine (STS), a potent and non-selective PKC inhibitor, had no significant effect on either. Another PLC inhibitor, U73122, but not its inactive analog, U73343, also inhibited PA evoked increases in [Ca2+]i. Depletion of extracellular calcium attenuated both basal and PA evoked increases in [Ca2+]i. The PLA(2) inhibitors, bromophenylacyl-bromide (BPB) and CDP-choline, had no effect on PA evoked [Ca2+]i responses. Neither the DAG analog, dioctanoylglycerol, nor the DAG kinase inhibitor, R59949, affected PA evoked changes in [Ca2+]i. Taken together, these data indicate that PA, in a manner independent of PKC, DAG, or PLA(2), may enhance Ca2+ release from IP(3) sensitive SR Ca(2+) stores via activation of PLC in neonatal rat cardiomyocytes. PMID- 10477829 TI - The impairment of essential fatty acid metabolism as a key factor in doxorubicin induced damage in cultured rat cardiomyocytes. AB - The clinical use of the antitumoral doxorubicin (DOX) is limited by its cardiotoxicity, which is mediated through different mechanisms. The membrane lipid peroxidation induced by DOX may cause disruption of the unsaturated fatty acyl chains; in the endoplasmic reticulum, containing the system catalyzing the desaturation/elongation of fatty acids, DOX could interfere with the metabolism of linoleic and alpha-linolenic acids. Using primary cultures of neonatal rat cardiomyocytes we demonstrated that the exposure to different concentrations of DOX (10(-5) and 10(-7) M) for 24 h caused an increase in the production of conjugated dienes, an impairment in the desaturation/elongation of essential fatty acids, and a reduction in the cellular content of highly unsaturated fatty acids. Conversely, 1 h exposure to 10(-5) M DOX was sufficient to induce alterations in the desaturation/elongation of linoleic and alpha-linolenic acids, but did not cause either formation of conjugated dienes or modification of the fatty acyl pattern. Therefore, DOX has a dual negative effect, depending on its concentration and on the time of exposure, one directed against the membrane highly unsaturated fatty acids, the other against the system which is required for the synthesis of these fatty acids themselves. These two effects synergically act in causing heart cell damage. PMID- 10477830 TI - Binding of phosphatidylinositol-specific phospholipase C to phospholipid interfaces, determined by fluorescence resonance energy transfer. AB - Dissociation constants for binding of phosphatidylinositol-specific phospholipase C from Bacillus cereus (bcPI-PLC) and the mammalian phosphatidylinositol-specific phospholipase C-delta(1) to lipid interfaces containing phosphoinositol, phosphocholine, and phosphomethanol head groups were determined by fluorescence resonance energy transfer. Dansyl-labeled lipid probes were used as acceptors, with intrinsic tryptophan of the enzyme as the donor. Titration of protein into lipid provided data from which K(d) and N, the limiting number of lipid molecules per protein bound, were calculated by non-linear regression analysis of exact binding equations. These results were compared with apparent K(m) values from kinetic data. K(d) values in the low microM range in terms of lipid monomers or low nM range in terms of binding sites were calculated with good fits of experimental data to theoretical binding curves. bcPI-PLC binds with high affinity to PI interfaces, slightly lower affinity to PC interfaces, and much lower affinity to PM interfaces. The mammalian enzyme also binds with high affinity to PI interfaces, but shows little or no binding with PC interfaces under similar concentration conditions. These K(d) values correlate reasonably with apparent K(m) values from kinetic experiments. PMID- 10477831 TI - Down-regulation of liver and heart specific fatty acid binding proteins by endotoxin and cytokines in vivo. AB - Fatty acid binding proteins (FABPs) are abundantly present in tissues that actively metabolize fatty acids (FA). While their precise physiological function is not known, FABPs have been shown to play a role in the uptake and/or utilization of FA within the cell. FA metabolism is markedly altered during the host response to infection and inflammation. Previous studies have demonstrated that endotoxin or bacterial lipopolysaccharide (LPS) enhances hepatic FA synthesis and re-esterification while inhibiting FA oxidation in liver, heart and muscle. Now, we have examined the in vivo effects of LPS and cytokines on FABPs in liver (L-FABP), heart and muscle (H-FABP). Syrian hamsters were injected with LPS, tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) and the mRNA and protein content for L-FABP and H-FABP were analyzed. 16 h after administration, LPS (100 microg/100 g body weight) produced a 72% decrease in L FABP mRNA levels in liver and this effect was sustained for 24 h. LPS also produced a 41% decrease in the protein content of L-FABP in liver after 24 h of treatment. TNF-alpha and IL-1beta decreased L-FABP mRNA levels in liver by 30 and 45%, respectively. LPS decreased H-FABP mRNA levels in skeletal muscle by 60% and in heart by 65%. LPS also produced a 49% decrease in H-FABP protein content in muscle. Neither TNF-alpha nor IL-1beta had any significant effect on H-FABP mRNA expression in heart and muscle. Taken together, these results indicate that LPS decreases FABP mRNA and protein levels in liver, heart and muscle, tissues that normally utilize FA as their primary fuel, whereas the inhibitory effect of cytokines is limited to the liver. The LPS-induced decrease in L-FABP and H-FABP may be an additional mechanism contributing to the decrease in FA oxidation that is associated with the host response to infection and inflammation. PMID- 10477833 TI - Correction: Case 20-1999. PMID- 10477832 TI - PGHS-2 inhibitors, NS-398 and DuP-697, attenuate the inhibition of PGHS-1 by aspirin and indomethacin without altering its activity. AB - Since the discovery of the inducible form of prostaglandin (PG) H synthase (PGHS), PGHS-2, considerable effort has been made to design selective inhibitors of this isozyme. N-(2-cyclohexyloxy-4-nitrophenyl) methanesulfonamide (NS-398) and 5-bromo-2-(4-fluorophenyl)-3-(4-methylsulfonyl) thiophene (DuP-697) have been shown to interact reversibly with PGHS-1, while irreversibly inhibiting PGHS-2 in a time-dependent manner. In the present study we have tested the effects of DuP 697 and NS-398 on the activity of PGHS-1 and further explored the interactions between these agents and the inhibition of PGHS-1 by aspirin, indomethacin and ibuprofen. Three independent experimental systems, namely bovine aortic endothelial cells (BAEC), human fibroblasts and ram seminal vesicle microsomes were used to investigate the effects of DuP-697 and NS-398 on PGHS-1. The results show that DuP-697 and NS-398, at concentrations ranges which do not inhibit PGHS 1 activity, significantly attenuated the inhibition of PGHS-1 that was caused by aspirin and indomethacin. The same concentrations of DuP-697 and NS-398 did not affect the inhibition of PGHS-1 that was induced by the competitive reversible inhibitors ibuprofen and naproxen. Similar effects of DuP-697 and NS-393 were obtained with ram seminal vesicle microsomes. These results suggest that PGHS-2 inhibitors DuP-697 and NS-398 possibly interact with PGHS-1 at a site different from the enzyme's catalytic site, thus causing attenuation of PGHS-1 inhibition by aspirin and indomethacin without altering PGHS-1 basal activity or the ibuprofen-induced inhibition. PMID- 10477834 TI - Use of older donor livers is associated with more extensive ischemic damage on intraoperative biopsies during liver transplantation. AB - Initial poor graft function is associated with increased morbidity and graft loss after liver transplantation. Donor age is a risk factor for the development of initial poor function. The severity of ischemic damage on intraoperative postreperfusion (0Post) allograft biopsy specimens is predictive of subsequent initial poor function. This study was performed to assess whether donor age is a risk factor for the development of ischemic damage on 0Post biopsy specimens. The records of 94 liver transplantations were reviewed. 0Post biopsy specimens were obtained after complete allograft revascularization. The severity of ischemic damage was graded as follows: 0, none; 1, minimal; 2, mild; 3, moderate; and 4, severe. Grafts were defined as older when donor age was 50 years or older. Other independent variables examined included donor cause of death, length of hospital stay, acidosis, serum alanine aminotransferase level, graft cold ischemia time, and degree of steatosis. Older grafts were associated with higher grades of ischemic damage than younger grafts (2.3 +/- 1.0 v 1.3 +/- 1.1; P =.003). Univariate and multivariate analysis identified donor age of 50 years or older as the only significant predictive variable of the severity of ischemic damage. In 16 transplantations involving older grafts, there was no statistically significant association between the severity of ischemic damage and incidence of initial poor function and graft loss. The use of older liver grafts is associated with more extensive ischemic damage immediately after graft reperfusion. Whether this early lesion identifies among older graft recipients those at risk for a worst outcome remains to be determined. PMID- 10477835 TI - Progressive splenomegaly after epoprostenol therapy in portopulmonary hypertension. AB - Patients with end-stage liver failure, portal hypertension, and associated pulmonary artery hypertension (portopulmonary hypertension [PPHTN]) have a high mortality when undergoing liver transplantation. Successful transplantation in these patients may depend on efforts to reduce pulmonary artery pressure (PAP). To this end, a number of centers are using a continuous intravenous (IV) infusion of epoprostenol, which has been shown to improve symptoms, extend life span, and reduce PAP in patients with primary pulmonary hypertension. We report four cases in which treatment of patients with PPHTN with continuous IV epoprostenol was followed by the development of progressive splenomegaly, with worsening thrombocytopenia and leukopenia. This finding may limit the usefulness of epoprostenol in PPHTN and influence the timing of transplantation in such patients. PMID- 10477836 TI - Standard liver volume in the Caucasian population. AB - Living donor and split-liver transplantation techniques require the calculation of a standard liver volume (SLV) as a reference point for the minimal volume necessary for the recipient. We therefore examined whether a widely used formula developed on the basis of a Japanese population sample was also adequate for the Caucasian population. The documentation of volumes of 1332 autopsy livers from a German Forensic Medicine Department was used to create a formula for an SLV for the Caucasian population. The Japanese formula estimated the Caucasian liver volume to be on average 322.6 +/- 335. 8 g (SD) less than they actually were. The following new formula for the calculation of SLV for Caucasians was established by linear regression analysis: Liver volume (mL) = 1072.8 * body surface area(m2) 345.7. PMID- 10477837 TI - Long-term follow-up after liver transplantation in patients with hepatic iron overload. AB - Patients with hepatic iron overload who undergo orthotopic liver transplantation (OLT) have a worse 1-year survival than those who undergo transplantation for other indications; the long-term outcome in this population is unknown. The purpose of this study is to report long-term follow-up after OLT in a cohort of patients with hepatic iron overload. Five liver transplant centers in the United States reported follow-up data on 37 patients receiving a first liver transplant who had severe hepatic iron overload in their native livers. Kaplan-Meier 5-year survival among these patients was compared with survival data from all age matched liver transplantations reported to the United Network for Organ Sharing (UNOS) over the same time period (1987 to 1993). The 5-year survival rate after OLT was 40% in the hepatic iron overload group compared with an overall survival rate of 62% for all patient groups from the UNOS registry (P =.0009). Although sepsis was the cause of 53% of all deaths occurring within the first year after OLT, cardiac complications accounted for 50% of the late mortality in patients with hepatic iron overload. In conclusion, long-term survival after OLT is significantly decreased in patients with hepatic iron overload. Infectious and cardiac complications are the most common causes of death in these patients. Further studies are needed to define the relationship between hepatic iron overload and mortality and to examine the effect of iron depletion on outcome after OLT in this patient population. PMID- 10477838 TI - Successful withdrawal of prednisone after adult liver transplantation for autoimmune hepatitis. AB - Corticosteroid withdrawal after orthotopic liver transplantation (OLT) represents an attractive therapeutic option for ameliorating post-OLT metabolic complications, although several reports suggest patients who undergo transplantation for autoimmune hepatitis (AIH) may have a greater incidence of acute and chronic rejection when withdrawn from corticosteroid therapy. The aim of this study is to evaluate the success of corticosteroid withdrawal in patients with AIH after OLT. Twenty-six patients underwent successful OLT for AIH. In 21 of these patients, stable maintenance immunosuppression consisted of cyclosporine (CSA) and prednisone (n = 20) or tacrolimus (TAC) and prednisone (n = 1). In this group, a trial of prednisone withdrawal was initiated when patients were 6 months or more post-OLT, with normal liver function, and receiving an average prednisone dosage of 10 mg/d. Five additional patients treated with either TAC (n = 4) or CSA (n = 1) plus mycophenolate mofetil underwent a 14-day taper of prednisone. Overall, 17 of 25 patients (68%) were successfully withdrawn from corticosteroids, with a mean follow-up of 22 months (range, 1 to 34 months). Of the remaining 8 patients, 5 patients received a lower dosage of prednisone or required prednisone to control inflammatory bowel disease. Only 3 patients remained dependent on prednisone to maintain stable liver allograft function. Withdrawal from 10 to 5 mg of prednisone (n = 21) resulted in four episodes of steroid-responsive and two episodes of steroid-resistant rejection in 3 patients, and 18 of 21 patients (86%) were rejection free. Withdrawal from 5 to 0 mg prednisone (n = 17) resulted in eight episodes of steroid-responsive and no episodes of steroid-resistant rejection in 4 patients; 13 of 17 patients (76%) were rejection free. Of the 5 patients in the 14-day prednisone-taper group, 3 patients had steroid-responsive rejection and 1 patient required OKT3. Seventeen of 21 patients (81%) with AIH were successfully withdrawn from corticosteroids. It is notable that corticosteroid withdrawal was associated with a reduction in serum cholesterol levels, decreased use of antihypertensive agents, and reduced need for insulin or oral hypoglycemic agents. We propose corticosteroid withdrawal should be attempted in patients with underlying AIH who undergo OLT because most will benefit without significantly jeopardizing the liver allograft. PMID- 10477839 TI - Inhaled nitric oxide reduces pulmonary artery pressures in portopulmonary hypertension. AB - Pulmonary artery hypertension in association with liver failure (portopulmonary hypertension [PPHTN]) is a significant barrier to liver transplantation because patients with this condition have a very high mortality when transplantation is undertaken. Inhaled nitric oxide (NO), a potent pulmonary vasodilator, reduces pulmonary artery pressure (PAP) in some patients with primary pulmonary hypertension, but its effect in patients with PPHTN is controversial. We investigated the hemodynamic effects of inhaled NO in 6 patients with PPHTN. Five of 6 patients responded to NO inhalation with decreases in PAP and pulmonary vascular resistance of greater than 10%; these decreases were statistically significant at NO concentrations of 10 and 30 ppm. Cardiac output did not significantly change. We conclude that inhalation of NO reduces PAPs in some patients with PPHTN. PMID- 10477840 TI - Evolving concepts in the diagnosis, pathogenesis, and treatment of chronic hepatic allograft rejection. AB - Chronic hepatic allograft rejection is characterized by the histological findings of ductopenia and a decreased number of hepatic arteries in portal tracts in the presence of foam cell (obliterative) arteriopathy. Recent studies have extended the histological spectrum of chronic rejection to include the presence of biliary epithelial atrophy or pyknosis involving the majority of small ducts present in the liver biopsy specimen. Overall, the incidence of chronic rejection in adults appears to be decreasing and is currently approximately 4%. However, the incidence of chronic rejection in pediatric liver transplant recipients has been more stable and ranges from 8% to 12% in most studies. Clinical risk factors associated with chronic rejection include: underlying liver disease, HLA donor recipient matching, positive lymphocytotoxic cross-match, cytomegalovirus infection, recipient age, donor-recipient ethnic origin, male donor into female recipient, number of acute rejection episodes, histological severity of acute rejection episodes, low cyclosporine trough levels, and retransplantation for chronic rejection. Chronic rejection, once diagnosed, frequently leads to graft failure; however, a number of reports indicated 20% to 30% of the patients with this diagnosis may respond to additional immunosuppressive therapy or even resolve spontaneously receiving baseline immunosuppression. Newer immunosuppressive agents, such as tacrolimus and mycophenolate, may successfully reverse chronic rejection, particularly when it is diagnosed in its early histological stages. PMID- 10477841 TI - Outcome of liver retransplantation in children. AB - Irreversible liver graft failure is a life-threatening complication. We reviewed the first 200 pediatric liver transplantations in Birmingham. Forty-one children developed primary graft failure, 9 of whom developed secondary graft failure. The main indications for graft failure were primary nonfunction (PRNF; 8 patients), vascular complications (VASC; 23 patients), and chronic rejection (CHRE; 19 patients). Thirty-two children underwent retransplantation (ReTx) (21 children received reduced grafts; 11 children, whole hepatic grafts). Patient survival was significantly worse for retransplant recipients compared with children receiving a single graft (63% v 76. 5% actuarial patient survival at 1 year; P <.05). Primary graft 1-year actuarial survival was 74% in first grafts compared with 47% for regrafts (P <.05), but improved with time. The graft 1-year survival rate was 55% for whole grafts and 45% for reduced and/or split grafts in the first 100 grafts compared with 83% and 66% in the second 100 grafts, respectively (P <.01). Emergency ReTx within a month of transplantation was associated with more complications and a worse outcome (1-year survival rate, 37%) compared with patients who underwent ReTx later (1-year survival rate, 72%; P <. 01). The incidence of primary graft failure decreased from 33% in the first 100 grafts to 16% in the second 100 grafts (P <.01), as did the incidence of PRNF, which decreased from 8% to 0% (P <.05). Although the rates of graft failure from VASC decreased from 15% to 8% (P =.2) and CHRE decreased from 11% to 8% (P =.6), neither reached statistical significance. The improved results overall are because of advances in surgical techniques, intensive care management, and graft preservation and refinements in immunosuppression. We conclude that ReTx for a child with primary graft failure is justified. PMID- 10477842 TI - Evidence of continuing bone recovery at a mean of 7 years after liver transplantation. AB - Patients with end-stage liver disease have low bone-turnover osteoporosis, and there is often further bone loss of 20% to 30% after orthotopic liver transplantation (OLT). Bone recovery after OLT has been reported, but data are limited. We undertook studies to determine whether bone recovery continues in the long term. Twenty-eight adult patients alive at least 5 years after OLT were studied (14 men, 14 women). Bone mineral density (BMD), serum parathyroid hormone (PTH), osteocalcin, and vitamin D levels were measured pretransplantation, at 3 months, 12 months, a mean of 46 months, and a mean of 85 months (range, 63 to 117 months) after transplantation. When BMD is expressed as a z score, the results were as follows: x0.82 +/- 0.22 pre-OLT; -2.04 +/- 0.27 at 3 months; -1.68 +/- 0.24 at 12 months; -1.23 +/- 0.24 at a mean of 46 months; and -1.0 +/- 0.26 at a mean of 85 months after OLT. The results at 46 and 85 months were significantly greater than the measurement at 3 months after OLT (P <.05). Furthermore, mean BMD (expressed as a z score) returns to the pre-OLT level at a mean of 85 months. At final follow-up, 9 of 28 patients had elevated PTH levels, and 14 of 27 patients had elevated osteocalcin levels. Five patients had spontaneous fractures in the first 12 months after transplantation, and 5 more patients had fractures by final follow-up. Even at 7 years after OLT, there was a significant increase in BMD (expressed as a z score) compared with 3 months after transplantation. Elevation of serum PTH and osteocalcin levels in some patients suggests continuing bone remodeling. PMID- 10477843 TI - Conversion of liver transplant recipients on cyclosporine with renal impairment to mycophenolate mofetil. AB - The management of liver transplant recipients with renal function impairment remains controversial because cyclosporine withdrawal from triple immunosuppression regimens may be followed by graft rejection. A nonnephrotoxic and powerful immunosuppressant such as mycophenolate mofetil (MMF) could allow a reduction of cyclosporine dosage or its withdrawal and an improvement in renal function in these patients. Eleven patients with serum creatinine levels greater than 1.5 mg/dL, normal graft function, and a rejection-free period of at least 1 year started MMF at a dose of 2000 mg/d (reduced in case of adverse events) while cyclosporine dosage was slowly reduced. At last follow-up (63 +/- 5 weeks), 7 patients remained free of cyclosporine (6 of those patients are also free of steroids), 2 patients reduced their cyclosporine dose, and 2 patients developed mild acute rejection that responded to a switch to tacrolimus therapy. Serum creatinine and urea levels in the 7 patients free of cyclosporine decreased from 2.22 +/- 0.13 to 1.90 +/- 0.19 mg/dL (P =.05) and 0.95 +/- 0.10 to 0.60 +/- 0.10 g/L (P <.001), respectively. Creatinine clearance increased from 38.16 +/- 5.60 to 47.01 +/- 6. 76 mL/min (P =.005). Control of arterial hypertension also improved. Tolerance to MMF was good, but 6 patients required dose reductions, mainly because of asymptomatic anemia. In conclusion, in liver transplant recipients with stable graft function, MMF may allow cyclosporine dose reduction or discontinuation, thus improving renal function and the control of arterial hypertension. This change of treatment must be carefully monitored because of the frequent need for MMF dose reduction and the risk for rejection. PMID- 10477844 TI - Influence of high donor serum sodium levels on early postoperative graft function in human liver transplantation: effect of correction of donor hypernatremia. AB - Donor hypernatremia was reported to cause postoperative graft dysfunction in human orthotopic liver transplantation (OLT). However, the effects of the correction of donor hypernatremia before organ procurement have not been confirmed. The aim of this study is to determine whether donor hypernatremia is associated with early graft dysfunction after OLT and to determine the effect of the correction of donor hypernatremia. One hundred eighty-one consecutive OLTs performed between May 1997 and July 1998 were entered onto this study. The cases were divided into three groups according to the donor serum sodium concentration: group A, serum sodium of 155 mEq/L or less before organ procurement (n = 118); group B, peak sodium greater than 155 mEq/L and final sodium 155 mEq/L or less (n = 36); and group C, final sodium greater than 155 mEq/L (n = 27). Graft survival within 90 days after OLT and early postoperative graft function were analyzed. There were no significant differences in donor and recipient variables among the three groups. The frequencies of graft loss were 15 of 118 grafts (12.7%) in group A, 4 of 36 grafts (11.1%) in group B, and 9 of 27 grafts (33.3%; P <.05 v groups A and B) in group C. The liver enzyme values in groups B and C were significantly greater than those in group A postoperatively. The prothrombin times of group C were significantly longer than those of group A for the first 4 postoperative days. Recipients of hepatic allografts from donors with uncorrected hypernatremia had a significantly greater incidence of graft loss compared with recipients of hepatic allografts from normonatremic donors. However, the differences in graft survival were abrogated by the correction of donor hypernatremia before procurement. PMID- 10477845 TI - Hyperbaric oxygen therapy for hepatic artery thrombosis after liver transplantation in children. AB - Early hepatic artery thrombosis (HAT) after pediatric orthotopic liver transplantation (OLT) can cause significant morbidity and mortality, leading to liver failure or septic complications requiring urgent retransplantation. Experimental evidence that hyperbaric oxygen (HBO) may ameliorate hepatic ischemic-reperfusion injury led to this study of HBO in pediatric liver transplant recipients who developed HAT. Children undergoing OLT under primary tacrolimus immunosuppression and University of Wisconsin organ preservation between August 1, 1989, and December 31, 1998, who developed HAT were the basis for this study. Patients who developed HAT between March 1, 1994, and December 31, 1998, were treated with HBO therapy until signs of ischemia resolved (absence of fever, normalizing liver injury test results) or for 2 weeks. The pediatric OLTs performed from August 1, 1989, to February 28, 1994, who developed HAT served as a control group. Primary outcome measures were survival, retransplantation rate, time to retransplantation, incidence of hepatic gangrene, and days to collateral formation. Three hundred seventy-five consecutive pediatric patients underwent 416 OLTs between August 1, 1989, and December 31, 1998. Thirty-one patients (7.5%) developed HAT at a mean time of 8.2 days (range, 1 to 52 days) post-OLT. In 17 patients, HBO treatment was begun within 24 hours of HAT or immediately after the revascularization attempt and performed twice daily for 90 minutes at 2.4 atmospheres pressure. Fourteen patients were treated without HBO. None of the HBO-treated patients developed hepatic gangrene. Eight HBO patients (47%) were bridged to retransplantation at a mean time of 157 days (range, 3 to 952 days) after initial OLT and all survived. Mean time to retransplant in the control group was 12.7 days (range, 1 to 64 days). HBO was well tolerated without significant complications. Although there was no significant difference in survival or retransplantation rates, HBO significantly delayed retransplantation, potentially by hastening the development of hepatic artery collaterals. PMID- 10477846 TI - Reducing the shortage of donor livers: what would It take to reliably split livers for transplantation into two adult recipients? AB - This article examines the scientific, technical, and administrative barriers to splitting donor livers for use in two adults. The main scientific barrier is that cadaveric donor livers at their current level of postoperative function are not sufficiently large to support life in two adult recipients. However, glycogenation of livers from young donors may be a method to overcome this problem in the short term. The three technical obstacles to splitting the liver in the midplane are anatomic anomalies that complicate or prevent splitting, the means to detect these anomalies, and the surgical methods to accomplish the split. Anatomic anomalies affecting the biliary drainage and arterial supply of the liver are the most important limiting technical factors. Administrative accommodations in the current methods of organ allocation will be needed if split liver transplantation in adults is to succeed. A nationwide view of organ allocation requires that the total number of lives saved by the procedure be the priority outcome nationally. If liver transplantation is viewed from this perspective, split-liver transplantation for adults would be a high priority, and incentives should be set to encourage it. PMID- 10477847 TI - Late recurrence and rapid evolution of severe hepatopulmonary syndrome after liver transplantation. AB - Recurrence of hepatopulmonary syndrome (HPS) after orthotopic liver transplantation (OLT) in an adult has never been reported. We describe a 23-year old woman who initially underwent OLT because of debilitating and severe HPS associated with nonalcoholic steatohepatitis (NASH). Although the clinical resolution of HPS was well documented day 117 post-OLT, the reappearance of NASH was documented by liver biopsy. Severe hypoxemia because of recurrent HPS rapidly evolved beginning approximately day 700 post-OLT. Retransplantation was attempted, but the patient died post-OLT of sepsis and/or multiorgan failure. PMID- 10477848 TI - Encephalopathy associated with human herpesvirus 6 in a liver transplant recipient. AB - Recent reports have documented human herpesvirus 6 (HHV-6) as a cause of high fever, bone marrow depression, and rash in liver transplant recipients in the absence of another known pathogen. We describe a 49-year-old liver transplant recipient who developed confusion, occipital headache, and involuntary movements of the limbs 3 weeks after orthotopic liver transplantation. HHV-6 was detected in the peripheral blood using a rapid culture assay. Examination of cerebrospinal fluid by polymerase chain reaction for HHV-6 was also positive. No other pathogens were identified. The patient improved after commencement of intravenous ganciclovir therapy. This case suggests HHV-6 needs to be considered in the differential diagnosis of unexplained confusion in liver transplant recipients. PMID- 10477849 TI - Pulmonic valve endocarditis after orthotopic liver transplantation. AB - Infective endocarditis is a rare complication affecting solid-organ transplant recipients. Isolated pulmonic valve endocarditis is also rare. A case of persistent bacteremia secondary to an isolated pulmonic valve vegetation occurred in a woman 10 days after liver transplantation. A pulmonary vegetectomy was performed as an alternative to valve replacement in addition to long-term antibiotic therapy. PMID- 10477850 TI - Predicting outcome after liver transplantation. PMID- 10477851 TI - The immunoreactive propensity of autoimmune hepatitis: is It corticosteroid dependent after liver transplantation? PMID- 10477852 TI - A laminin and nerve growth factor-laden three-dimensional scaffold for enhanced neurite extension. AB - Agarose hydrogel scaffolds were engineered to stimulate and guide neuronal process extension in three dimensions in vitro. The extracellular matrix (ECM) protein laminin (LN) was covalently coupled to agarose hydrogel using the bifunctional cross-linking reagent 1,19- carbonyldiimidazole (CDI). Compared to unmodified agarose gels, LN-modified agarose gels significantly enhanced neurite extension from three-dimensionally (3D) cultured embryonic day 9 (E9) chick dorsal root ganglia (DRGs), and PC 12 cells. After incubation of DRGs or PC 12 cells with YIGSR peptide or integrin beta1 antibody respectively, the neurite outgrowth promoting effects in LN-modified agarose gels were significantly decreased or abolished. These results indicate that DRG/PC 12 cell neurite outgrowth promoting effect of LN-modified agarose gels involves receptors for YIGSR/integrin beta1 subunits respectively. 1,2-bis(10, 12-tricosadiynoyl)-sn glycero-3-phosphocholine (DC(8,9)PC)-based lipid microcylinders were loaded with nerve growth factor (NGF), and embedded into agarose hydrogels. The resulting trophic factor gradients stimulated directional neurite extension from DRGs in agarose hydrogels. A PC 12 cell-based bioassay demonstrated that NGF-loaded lipid microcylinders can release physiologically relevant amounts of NGF for at least 7 days in vitro. Agarose hydrogel scaffolds may find application as biosynthetic 3D bridges that promote regeneration across severed nerve gaps. PMID- 10477853 TI - Effect of a poly(propylene fumarate) foaming cement on the healing of bone defects. AB - Regeneration of skeletal tissues has been recognized as a new means for reconstruction of skeletal defects. We investigated the feasibility of an injectable and expandable porous implant system for in situ regeneration of bone. Therefore, a composite biodegradable foaming cement based on poly(propylene fumarate) was injected into a critical size defect made in the rat tibia. Animals were divided into two groups comparing the foam in the experimental group against sham-operated animals having a drill hole but no implant in the control group. Eight animals were included in each group. Animals were sacrificed at 1, 3, and 7 weeks postoperatively. Implantation sites were then evaluated with histologic and histomorphometric methods. Results of this study showed that defects did not heal in sham-operated animals. In the experimental group, metaphyseal and cortical defects healed within the first postoperative week by formation of immature woven bone. At the site of the cortical drill hole defect, healing was noted to progress to complete closure by formation of mature bone. Histomorphometry corroborated these findings and showed that metaphyseal bone remodeling peaked at 1 week postoperatively and then decreased as healing of the cortical defect progressed. This suggests that near-complete restoration of the original state of the tibial bone occurred in this animal model supporting the concept of in situ bone regeneration by application of engineered biodegradable porous scaffolds. () () PMID- 10477854 TI - Biomechanical analysis of a chondrocyte-based repair model of articular cartilage. AB - The objective of this study was to evaluate the biomechanical properties of newly formed cartilaginous tissue synthesized from isolated chondrocytes. Cartilage from articular joints of lambs was either digested in collagenase to isolated chondrocytes or cut into discs that were devitalized by multiple freeze-thaw cycles. Isolated cells were incubated in suspension culture in the presence of devitalized cartilage matrix for 3 weeks. Multiple chondrocyte/matrix constructs were assembled with fibrin glue and implanted subcutaneously in nude mice for up to 6 weeks. Testing methods were devised to quantify integration of cartilage pieces and mechanical properties of constructs. These studies showed monotonic increase with time in tensile strength, fracture strain, fracture energy, and tensile modulus to values 5-10% of normal articular cartilage by 6 weeks in vivo. Histological analysis indicated that chondrocytes grown on dead cartilage matrix produced new matrix that integrated individual cartilage pieces with mechanically functional tissue. PMID- 10477855 TI - Meniscus regeneration in a rabbit partial meniscectomy model. AB - Meniscectomy is known to be associated with osteoarthrosis of the knee. The purpose of this study was to compare the natural and augmented repair of menisci in the knees of New Zealand White rabbits. To create a partial defect in the medial meniscus, we used an experimental model that has been well characterized and extensively used in the study of osteoarthrosis and articular cartilage repair. The defect was left untreated or treated with one of the following: a periosteal autograft, a type I collagen sponge, or the same sponge loaded with autologous, bone marrow-derived, cultured mesenchymal stem cells. The natural repair was always incomplete and degenerative changes within these joints were progressive. The periosteal autograft underwent differentiation into a bone and hyaline cartilage composite that was ineffectual as a meniscus and accelerated the degenerative changes in those joints when compared to natural repair controls. There was evidence of a consistent sequence of events in the transformation of the periosteal grafts to a core of cartilage that underwent endochondral ossification. In the last two groups, the collagen sponge functioned as a scaffold that resulted in more abundant repair tissue. The collagen sponge alone supported a largely fibrous repair process. The cultured mesenchymal stem cells were observed to augment the repair process in some specimens to include fibrocartilage histologically similar to normal meniscus. Degenerative changes were present in both of these groups, which indicates that the biomechanical function of the meniscus was not restored, or an irreversible osteoarthrosis cascade was initiated during the repair period. Based on these preliminary studies, further investigation of cell-based meniscus regeneration appears to be warranted. PMID- 10477856 TI - End-to-end anastomosis between tissue-engineered intestine and native small bowel. AB - The purpose of this study was to demonstrate the feasibility of end-to-end anastomosis between tissue-engineered intestine and native small bowel and to investigate the effect of this anastomosis on their growth. Microporous biodegradable polymer tubes were created from a fiber mesh of polyglycolic acid sprayed with 5% polylactic acid. Intestinal epithelial organoid units were harvested from neonatal Lewis rats and seeded onto polymers. These constructs were implanted into the omentum of adult Lewis rats. Three weeks after the implantation, the constructs (n = 7) were anastomosed to the native jejunum in an end-to-end fashion. Ten weeks after implantation, the tissue-engineered intestine was harvested. Four of 7 rats survived for 10 weeks and the overall patency rate of the anastomosis was 78% (11 of 14 anastomosis). The maximal length of the tissue-engineered intestine at week 3 and 10 was 1.80 +/- 0.32 and 1.93 +/- 0.39 cm (mean +/- SD). Histologically, the tissue-engineered intestine was lined with a well-developed neomucosal layer that was continuous with the native intestine. We conclude that anastomosis between tissue-engineered intestine and native small bowel had a moderately high patency rate and had a positive effect on maintenance of the size of the neointestine and development of the neomucosa. PMID- 10477857 TI - Metabolic flux analysis: a powerful tool for monitoring tissue function. AB - In recent years, metabolic flux analysis has been widely used in bioprocess engineering to monitor cell viability and improve strain activity. Metabolic flux analysis refers to a methodology for investigating cellular metabolism whereby intracellular fluxes are calculated using a stoichiometric model for the major intracellular reactions and applying mass balances around intracellular metabolites. A powerful feature of this methodology is its ability to consider cellular biochemistry in terms of reaction networks. By considering the stoichiometry of biochemical reactions, it is possible to estimate the degree of engagement of each pathway participating in overall cellular activity, and hence obtain a comprehensive view of a cell s metabolic state. Given the potential impact of cellular energy metabolism on the function of engineered tissues, such comprehensive analysis of metabolic activity can be an extremely useful tool for tissue engineers. Estimates of intracellular fluxes under various environmental conditions could be used to optimize function in vivo as well as culture conditions in vitro. In this review, we provide a brief theoretical background of metabolic flux analysis and summarize the most widely used experimental approaches to obtain flux data. This review is intended as an overview of the field and as a starting point for tissue engineers wishing to learn about and eventually employ this methodology. PMID- 10477858 TI - In vivo biocompatibility and degradation studies of polyhydroxyoctanoate in the rat: a new sealant for the polyester arterial prosthesis. AB - The present study examined the biocompatibility and degradation properties of poly (beta-hydroxy octanoate) (PHO) as an impregnation substrate on arterial prostheses. PHO-impregnated polyester grafts sterilized by ethylene oxide (EO) or gamma (gamma) radiation, and polyester Dacron(R) prostheses impregnated with fluoropolymer, gelatin, or albumin were implanted subcutaneously in rats for periods ranging from 2 to 180 days. The biocompatibility was assessed by quantifying the alkaline and acid phosphatase secretion while performing histological studies at the tissue/prosthesis interface. The degradation was determined by chemical analysis of the EO and gamma-sterilized PHO after implantation using differential scanning calorimetry (DSC), wide angle x-ray diffraction (WAXD), and size exclusion chromatography (SEC). Alkaline phosphatase activity by the sterilized PHO and by the gelatin and albumin grafts was significantly elevated early after implantation in contrast to that of the Dacron and fluoropolymer grafts that occurred later, at 7 and 5 days, respectively The peak of acid phosphatase activity for all of the grafts occurred between 5 and 10 days postimplantation, with the gamma-sterilized PHO grafts recording the greatest activity. Histological study revealed that the tissue incorporation into the graft wall was earlier and more complete for the Dacron and fluoropolymer grafts after 6 months than for the gelatin and albumin grafts, because the latter induced important inflammatory reactions during the resorption of the cross linked protein substrates. The EO and gamma-sterilized PHO grafts exhibited a similar healing sequence characterized by the development of a collagenous tissue surrounding the prostheses. However, no infiltration of tissue into the graft wall was observed after 6 months, mainly because of the presence of the PHO. Degradation of the EO and gamma-sterilized PHO occurred preferentially by a hydrolytic mechanism as shown by a 30% molecular weight decrease after 6 months. In conclusion, PHO showed good biocompatibility in terms of enzyme activity and tissue reaction. Degradation was a slow, in vivo process controlled primarily by a random hydrolytic reaction and by a local enzymatic attack by macrophages and giant cells. PMID- 10477859 TI - The role of vascular smooth muscle cell integrins in the compaction and mechanical strengthening of a tissue-engineered blood vessel. AB - Vascular smooth muscle cells (VSMC) influence vessel structure and function during normal development, and in disease states. VSMC interactions with extracellular matrix, via cell surface integrins, play an important role in these processes. A greater understanding of the molecular basis of these interactions is also critical to advances in the field of cardiovascular tissue engineering. This study examined the role of VSMC integrins in the spontaneous compaction and eventual strengthening of a rudimentary tissue-engineered blood vessel (TEBV) consisting of a fibrillar type I collagen network populated by human aortic smooth muscle cells. Using integrin subunit-specific antibodies, we demonstrated that anti-beta1 (Mab13 and P4C10) and anti-alpha2 (P1E6) antibodies that inhibit aortic smooth muscle cell (AoSMC) adhesion to collagen, also significantly inhibit TEBV compaction during the 24-hour period following TEBV construction. However, no difference in the tensile stress of antibody-treated and control TEBVs was observed at this time point. In contrast, 72 hours after construction, the inhibitory effect of anti-integrin antibodies on compaction had been overcome but tensile stress was decreased in TEBVs treated with anti-alpha2/anti-beta1 antibodies when compared to controls. These data provide evidence linking VSMC integrins, specifically the alpha2beta1 integrin, with the initial compaction, as well as, the postcompaction strengthening of the TEBV. PMID- 10477860 TI - [Length of work disability and associated factors]. AB - OBJECTIVE: To estimate the mean length of temporary work disability (TD) and to assess some of its possible determinants. METHODS: During 1995, we registered 12,570 cases of TD among people from the Inspection Area of Alcala of Henares. We obtained a random sample of 600 cases. In addition to TD length, we obtained information about patients' sociodemographic characteristics, social security group, diagnosis, and type of primary care. Analysis of variance was used to compare means and Cox regression models were used to estimate risk ratios for factors influencing return to work. RESULTS: The mean and the median of the TD were 44.5 and 11 days respectively. The TD length was less than one month in 72.9% of the cases. The TD length was similar in both sexes and showed a positive association with age (p=0.0002). The General group had higher TD length than the Insalud group and lower than the Autonomous and Housework groups. The Insalud personnel showed an adjusted relative effect (and 95% confidence intervals) in the probability of returning to work of 2 (1.07-3.73) compared to the General group. The adjusted relative risk was 0.89 (0.88-0.90) for an increase of 5 years of age. CONCLUSIONS: The most important factors in the TD length were age and social security group. The improvement in the health system accessibility is one of the factors that may positively affect the probability of returning to work. PMID- 10477861 TI - [Temporary disability: analysis strategies]. AB - OBJECTIVES: To apply different regression models to estimate rate ratios for temporary sick-leave (TSL) which may occur several times in the same individual during a period, and the frequency is not constant for the observation period. SUBJECTS AND METHODS: All workers employed more than 30 days between January 1st of 1992 and June 1st of 1995 were included into the population study. The following period was 1,259 days and the total number of workers included in the study was 2,306. During that period 2,649 TSL episodes were notified, which meant 85,947 lost days. Poisson regression, Generalised Estimating Equations (GEE) and Andersen-Gill modification of Cox regression modify by Wei (WLW) were applied. RESULTS: The highest TSL incidence rates were seen in women, lesser than 30 years old, cleaners, maintenance workers and auxiliary nurses, and those involved in shiftwork. This profile was not modified after applying GEE and WLW regression models, although confidence intervals were widened. CONCLUSIONS: TSL data does not fit Poisson regression assumptions, but GEE and WLW regression models do not appear as alternatives. Other conditional regression models would need to be explored to suitably analyse this data. PMID- 10477862 TI - [Validity of a job stressors measurement scale in nursing staff: the nursing stress scale]. AB - BACKGROUND: The aim of this work was to determine the reliability and construct validity of a scale intended to measure job stressors to which hospital nursing staff may be exposed. SUBJECTS AND METHODS: The nursing stress scale contains 34 stressors. The scale's trans-cultural adaptation was carried out by means of the translation-back translation method. Validation was conducted on a random sample of 201 health professionals in a public hospital in Valencia. The reliability of the scale was assessed after its readministration on a sub-sample of 30 nursing professionals, with a 15 day interval. The construct validity was obtained through the correlation of the scale with another two scales: The 28 item version of Goldberg's General Health Questionnaire and 7 dimensions of the Health questionnaire SF-36. RESULTS: The scale in Spanish language contains 34 items after eliminating the first item from the original scale (N1: computer failure) and including a new item (E1: Frequent job interruptions). The distribution of scores obtained in the initial administration of the scale and fifteen days later do not differ statistically. The Cronbach's alfa coefficient is 0.92 for the total scale and in each of the sub-scales ranges between 0. 83 and 0.49. The correlation between the scale and the GHQ-28 items questionnaire is 0.34. For each one of the seven dimensions of the SF-36 questionnaire the correlations range between -0.21 and -0.31. CONCLUSION: The nursing stress scale is a useful instrument for measuring possible stressors in this collective. It has high internal consistency and construct validity, as does the original American version, however reliability is moderate. PMID- 10477863 TI - [Gender differences in the influence of housework on health]. AB - OBJECTIVE: To analyze the influence of domestic workload on the health of the working population of Catalonia, taking into account its differential impact on males and females. SUBJECTS AND METHODS: The study population were the 2,164 men and 1,215 women interviewed in the Catalonian Health Survey (ESCA) of 1994, with a paid work who were the main responsible for the family unit. The dependent variable was the self-perceived health status that was dichotomized for the bivariate and multivariate analysis. The domestic workload was measured with the number of people living at home (2, 3, 4 or more than 4), living with children younger than 15 and living with older than 65. The adjusting variables were age and occupational social class. The analysis was separated for men and women and crude, adjusted for age, and for all the predictor variables odds ratios (OR) were calculated with their 95% confidence intervals (95%CI). RESULTS: There was no relation between domestic workload and health status among men. Among women the risk or poor health status increased with the number of people living at home with adjusted OR associated to living with more than four persons of 2.35 (95%CI = 1. 33-4.15), as compared with living only with the spouse. Living with older than 65 was protector for women (adjusted OR = 0.63; 95%CI = 0. 39-1.03). CONCLUSIONS: The risk of poor health status associated to the domestic workload among women with paid work, suggests the need of increasing community resources for caring children, as well as, increasing the participation of men in the domestic work. Future health surveys should collect information about the number of people of different age strata living at home. PMID- 10477864 TI - [Social class differences in bladder cancer in Catalonia]. AB - OBJECTIVE: To assess the presence of socio-economic differences in bladder cancer in a heavily industrialised area of Catalonia, Spain and to evaluate risk factors associated with the presence of these differences. METHODS: 218 cases and 344 population controls resident in the area of Valles Occidental, Barcelona, were identified prospectively and interviewed during 1993-1995. Socio-economic level was evaluated through attained education and social class on the basis of the last and longest held occupation. Logistic regression was used to derive adjusted odds ratios and 95% confidence intervals. RESULTS: There was no association between education and bladder cancer risk. Social class, evaluated on the basis of longest held job, was weakly related with bladder cancer, with the highest risk occurring in the highest social classes. This pattern was more pronounced when the analysis was done using the last occupation. Occupational exposures modified minimally these results. Among controls, the overall prevalence of smoking did not differ between social class but the prevalence of heavy-smokers (> 48.76 packs-year) was highest among subjects in social class V (35.5%) and lowest in social class I (10%). Adjusting for smoking enlarged social class differences. CONCLUSION: Similarly to the pattern in other European countries, no pronounced differences by social class were observed for bladder cancer in this area of Catalonia. The evaluation of social class patterns was crucially affected by the type of socioeconomic information examined. Measures of life-time experience should be preferred rather than socio-economic level at the time of the disease. Smoking seems to be one of the main mediators for the occurrence of bladder cancer differentials but other, yet unidentified risk factors, must be associated with bladder cancer risk, especially among the highest social classes. PMID- 10477865 TI - [Public health structures in Spain: a changing panorama]. AB - In the last years the health care system in Spain has undergone very important transformations. However, the public health services have not followed a comparable process. Until 1979 public health structures were based in central services and their provincial units, on one side, and in the resources of local governments on the other. From then on began the process of transferring responsibilities and resources to the regional governments of Autonomous Communities (AC), which today manage most public health services, while the central government keeps as its responsibility the development of basic norms, the administration of public health services at borders and customs, and the general health coordination. The situation in 1995 resembles that of a Federal Country, although the Kingdom of Spain is not formally defined as such. The role of central government services is visibly reoriented towards coordination and the building of consensual and shared structures for health information, for need evaluation, and for policy formulation and evaluation. Although the General Health Law of 1986 considered public health as a main axis for all health agencies, its actual development has been more patchy. Several AC with an Autonomous Health Service have kept public health services separated from it. Besides, many public health functions and activities are developed today from other structures. The processes of change reveal two contradictory aspects. On one side, structures have been upgraded after decentralization, and teams reinforced, with trained and full-time staff. However, there is some dilution of public health responsibility in the periphery, and a lower visibility of the health authority. PMID- 10477866 TI - [A navigational guide for organizing and clarifying the health care debate]. AB - In this brief paper, discussion focuses on options currently under consideration for the reform of the Spanish health care system. After a brief review, readers are advised, in view of the consequences of each option, to consider all the arguments in order to make fully informed decisions. Finally, it is proposed that, in view of what is known and unknown about the effects of different health care policy options, a research agenda be found that avoids both complacency and categorical affirmations regarding this complex topic. Arguments are roughly presented as an algorithm. Simplifications are made for the sake of clarity and the analysis is qualitative. Each nexus of the algorithm covers the most relevant aspects of the economic and health care issues that each alternative involves. In the author's opinion, these points should be kept in mind when making decisions. PMID- 10477868 TI - Commentary PMID- 10477867 TI - Another view of the tamoxifen trial. PMID- 10477869 TI - Laparoscopic management of women with a family history of ovarian cancer. AB - BACKGROUND AND OBJECTIVE: The safety of laparoscopic surgery in women with a family history of ovarian cancer predicted to have benign disease has not been established. The objective of this study was to evaluate the feasibility and complications of operative laparoscopy and to describe the pathologic findings in this patient population. METHODS: Sixty-two consecutive women with a family history of ovarian cancer who elected prophylactic oophorectomy or had predicted benign adnexal disease were offered laparoscopic surgery. Patient characteristics, details of laparoscopic surgery, operative and postoperative complications, and histopathologic findings were recorded. RESULTS: Laparoscopy was converted to laparotomy in 2 patients (3.2%); 1 patient (1.6%) had an operative vascular complication, and 1 patient (1.6%) had postoperative bleeding. Median (range) estimated blood loss, operative time, and hospital stay were 50 ml (50-1,500), 120 min (60-290), and 1 day (0-9), respectively. Histopathologic findings included normal ovaries (n = 20), corpus luteum cyst (n = 16), follicular cyst (n = 8), endometriotic cyst (n = 7), serous cystadenoma (n = 5), epithelial hyperplasia (n = 4), dermoid cyst (n = 1), and fibrothecoma (n = 1). CONCLUSIONS: Laparoscopic surgery is feasible and safe in women with a family history of ovarian cancer predicted to have benign disease and is associated with low blood loss and short hospital stay. Most ovaries removed are either normal or have benign disease. PMID- 10477870 TI - Use of subatmospheric pressure to prevent doxorubicin extravasation ulcers in a swine model. AB - Application of subatmospheric pressure to sites injected with doxorubicin prevented ulcer formation in treated sites (0 ulcers/16 sites) compared to control wounds (10 ulcers/16 sites) in a pig model. BACKGROUND AND OBJECTIVES: Extravasation of doxorubicin hydrochloride (Adriamycin) frequently causes chronic ulcers, which usually progress and expose underlying structures such as tendons and bone. The exact mechanism of action that causes cell death and the chronic ulcers is unknown. METHODS: Eight sites were injected intradermally with doxorubicin on each of 4 pigs. Four sites on each animal served as untreated controls. The remaining four sites were exposed to 125 mm Hg subatmospheric pressure applied 1 h after injection. The sites were observed on a three times per week schedule. Sites that did not develop ulcers were re-injected up to a total of four injections. The animals were observed for 5 weeks. RESULTS: Ten of sixteen control sites developed ulcers. No subatmospheric pressure treated sites developed ulcers. The incidence of ulcer formation was significantly less for treated wounds compared to control wounds at P < 0.001 by Fisher's exact test. CONCLUSIONS: This physical modality appears to successfully prevent ulcer formation after doxorubicin injection. PMID- 10477871 TI - Identification of semaphorin E gene expression in metastatic human lung adenocarcinoma cells by mRNA differential display. AB - Human lung adenocarcinoma cell lines HAL-8Luc and HAL-24Luc differ in their metastatic potential. HAL-8Luc cells metastasize to lungs when injected either intravenously or intramuscularly. in mice while HAL-24Luc cells do not. The differential display method is used to identify genes differentially expressed between the two cell lines and the findings are extensively discussed. BACKGROUND: Lung cancer is the leading form of cancer in most countries, and metastasis is the main cause of death in oncological patients. The metastatic phenotype of tumor cells is the result of genetic events altering the RNA and protein expression of normal cells. Our objective was to identify genes expressed differentially between metastatic and nonmetastatic human lung adenocarcinoma cells that might be used as a prognostic factor. METHODS: The differential display technique was used to compare the RNA expression patterns distinguishing metastatic (HAL-8Luc) and nonmetastatic (HAL-24Luc) human lung adenocarcinoma cells, two genetically close cell lines. RESULTS: Differential expression of three cDNAs was confirmed by Northern blot analysis. Two sequences corresponding to a putative splicing factor and a proliferation-related factor cDNAs were underexpressed in the metastatic cells relative to the nonmetastatic ones. Interestingly, we found that human semaphorin E mRNA was several fold overexpressed in the metastatic cells. This recently identified gene encodes a protein whose expression has been related to several cell survival mechanisms as well as to immunosuppression. CONCLUSION: Our results point to the relevance of semaphorin E in metastatic spread of human lung adenocarcinoma cells. PMID- 10477873 TI - Commentary PMID- 10477872 TI - Initial experience in a community hospital with sentinel lymph node mapping and biopsy for evaluation of axillary lymph node status in palpable invasive breast cancer. AB - BACKGROUND AND OBJECTIVES: To determine the sentinel node detection rate and the accuracy with which the sentinel node histology reflects that of the axilla in a series of patients with palpable invasive breast cancer. METHODS: Forty-four patients with clinically node-negative palpable invasive T1 or T2 breast tumors underwent sentinel node biopsy using isosulfan blue dye, followed immediately by either local excision of the primary lesion with standard axillary lymph node dissection or modified radical mastectomy. All surgeries were performed at Northwest Hospital, Seattle, Washington, between January 1996 and October 1997. RESULTS: The sentinel node was successfully identified in 73% of the patients (32/44). The frequency of sentinel node detection was greater for tumors in the outer quadrants than the inner quadrants (z-test, P < 0.001). Of the 32 patients in whom a sentinel node was identified, 10 (31%) had histologically positive sentinel nodes: 5 (16%) by frozen section, 2 additional patients (6%) after permanent hematoxalin-eosin (H&E) stained sections, and the remaining 3 (9%) after immunohistochemical stains for cytokeratins when the FS and permanent H&E stained sections were benign. Twenty patients had benign axilla. The sentinel node was falsely negative in 2 patients, yielding an accuracy of 93.8%, sensitivity of 83.3%, and negative predictive value of 91%. CONCLUSIONS: Lymphatic mapping is technically feasible for patients with small (T1 or T2) palpable invasive breast tumors. The sentinel node can be reliably identified in the majority of these patients, and its histology reflects that of the axilla with a high degree of accuracy. Immunohistochemical stains and permanent H&E stained sections of the sentinel node increased the test's ability to correctly identify axillary metastases. Improving this sensitivity remains a primary goal, however, if benign sentinel node histology is to be used as a criterion to preclude axillary dissection. PMID- 10477875 TI - Commentary PMID- 10477874 TI - Case of retroperitoneal dedifferentiated mixed-type liposarcoma: comparison of proliferative activity in specimens from four operations. AB - In a case of retroperitoneal dedifferentiated mixed-type liposarcoma, a dedifferentiated component was observed in the so-called mixed-type liposarcoma consisting of well-differentiated and myxoid components. The proliferative activity was compared among the different components of the tumor by immunohistochemical study using the proliferating cell nuclear antigen (PCNA) and MIB-1 monoclonal antibodies. The dedifferentiated component showed higher positivity than the well-differentiated and myxoid components, and tumor progression was most advanced in the dedifferentiated component. In the chronological examination of each component, the labeling indices of PCNA and MIB 1 were significantly higher at the third recurrence than in the primary lesion in all types, indicating that the proliferative activity of the tumor cells increased gradually. Considering the surgical treatment of liposarcoma, an extended resection encompassing normal adjacent tissues is required in cases containing the dedifferentiated component in comparison to the cases containing only well-differentiated or myxoid components. PMID- 10477876 TI - Prevention of recurrent giant-cell tumors of long bones--a new surgical technique. PMID- 10477877 TI - Commentary PMID- 10477878 TI - Gastric transposition with pharyngogastric stapler anastomosis. PMID- 10477879 TI - Current status of tamoxifen use: An update for the surgical oncologist. AB - The surgical oncologist is frequently responsible for the screening and diagnosis of women with breast cancer. In this pivotal role, they are often the first to discuss treatment options, including nonsurgical interventions, with breast cancer patients. Recent long-term clinical trial data provide support for the use of tamoxifen to prevent breast cancer in women at high risk of the disease. A breast cancer risk assessment can help identify women at higher than average risk for the disease, who may be appropriate candidates for chemoprevention. It is important for the surgical oncologist to understand the current indications and evidence regarding the use of tamoxifen for breast cancer prevention and treatment as they counsel their patients on available options. PMID- 10477881 TI - Commentary PMID- 10477880 TI - Total vascular exclusion for liver resections: pros and cons. AB - Dramatic improvements in morbidity and mortality rates following liver resections have been reported in the past decade. Consequently, the indications for hepatectomy are becoming more liberal. Many techniques of liver resection with or without vascular clamping have been reported with excellent clinical results. Total vascular exclusion (TVE) of the liver during parenchymal transection has been advocated susceptible to increase the resectability of tumors that might not be safely approached by other techniques. Cirrhotic livers are probably more vulnerable to ischemic injury related to TVE than normal livers. The indications and technical and metabolic aspects of the technique are reviewed. PMID- 10477882 TI - Calf sonography for detecting deep venous thrombosis in symptomatic patients: experience and review of the literature. AB - PURPOSE: We determined the accuracy of sonography in the detection of isolated calf deep venous thrombosis (DVT) and the rate of indeterminate ultrasound examinations in patients with physical signs or symptoms suggestive of DVT. METHODS: We reviewed the medical literature (MEDLINE) to determine the accuracy of sonography and the frequency of indeterminate studies in detecting isolated calf DVT in patients with physical signs or symptoms suggestive of DVT. A meta analysis was used to derive summary measures of sensitivity, specificity, and accuracy from studies in which 5 or more isolated calf DVT were identified. Frequencies of indeterminate examinations were recorded for studies in which these data were provided, and we pooled these results with our own data for 196 patients. RESULTS: The meta-analysis revealed that sonography correctly identified isolated calf DVT in 49 of 53 extremities (sensitivity, 92.5%; 95% confidence interval, 81.8-97.9%) and correctly identified the absence of calf DVT in 157 of 159 extremities (specificity, 98.7%; 95% confidence interval, 95.5-99. 9%), yielding an accuracy of 97.2% (95% confidence interval, 93.9-99. 0%) for ultrasound examinations considered diagnostic. However, when evaluating our patient population and the literature, we found a substantial number of indeterminate studies (overall rate of 54.6% in 463 extremities), with a wide variation in the reported frequency of indeterminate studies (9.3-82.7%). CONCLUSIONS: Sonography is highly accurate in detecting isolated calf DVT in symptomatic patients, but indeterminate studies occur frequently, with a wide range of reported rates. Each ultrasound laboratory should evaluate its own rate of indeterminate studies. PMID- 10477883 TI - Sonographic detection of occult fractures in the foot and ankle. AB - PURPOSE: The purpose of this retrospective study was to determine whether high resolution sonography can aid in the diagnosis of radiographically occult fractures in the foot and ankle. METHODS: High-resolution sonography with a 10 MHz linear-array transducer was performed in 268 patients with foot and ankle injuries whose initial plain x-ray films were negative for fracture. RESULTS: Twenty-four patients had occult fractures demonstrated by sonography. On sonography, the occult fractures appeared as a discontinuity of cortex echogenicity. The fractures were found at the calcaneus (n = 8), metatarsus (n = 6), talus (n = 3), navicular bone (n = 3), cuboid bone (n = 2), cuneiform bone (n = 1), and lateral malleolus (n = 1). Review of the patients' radiographs revealed tiny fractures at the sonographically identified locations in 2 patients. The first 5 patients underwent bone scans, which confirmed the presence of the fractures. The first 11 patients received follow-up sonographic examination 6 weeks after diagnosis; in all 11, an echogenic line over the previous fracture site, presumably representing callus formation, was noted. CONCLUSIONS: Sonography-a readily available, noninvasive imaging technique-can provide important information about soft tissue injuries and cortical discontinuities in the foot and ankle area. Using this procedure, occult fractures can be identified and delineated, and costly procedures such as MRI can be avoided. PMID- 10477884 TI - Color doppler sonography avoids misinterpretation of the "parallel channel sign" in the sonographic diagnosis of cholestasis. AB - PURPOSE: The intrahepatic "parallel channel sign" on gray-scale sonograms is generally interpreted as representing dilated bile ducts, but it may also be caused by enlargement of intrahepatic arteries. This study was performed to evaluate the incidence of misinterpretation of the parallel channel sign without color Doppler sonography and the characteristics of patients in whom misinterpretation of the parallel channel sign is likely to occur. METHODS: A total of 1,100 patients were examined by sonography. All patients with a parallel channel sign on gray-scale sonograms underwent color Doppler sonography. In addition, laboratory values related to cholestasis were measured. RESULTS: The parallel channel sign was observed in 57 patients (5.2%). In 35 (61%) of these patients, color Doppler sonography revealed blood flow in both lumina, indicating that the parallel channel sign was not caused by enlarged bile ducts. Eighty-six percent of this group had nonbiliary liver disease; the enlarged hepatic artery branches were associated with liver cirrhosis in 63% of this group. Color Doppler sonography confirmed that the other 22 patients (39%) had an enlarged intrahepatic bile duct. CONCLUSIONS: Use of color Doppler sonography can help avoid misinterpretation of the parallel channel sign, especially in patients with nonbiliary liver disease. PMID- 10477885 TI - Effect of color Doppler system on the twinkling sign associated with urinary tract calculi. AB - PURPOSE: The aim of this study was to assess the effect of the color Doppler system on the artifact known as the "twinkling sign." METHODS: Sixty-five patients with at least 1 urinary tract calculus seen on sonography were examined with color Doppler sonography. Two color Doppler systems of 2 different generations were used. Examinations included gray-scale, color Doppler, power Doppler, and spectral evaluations of the calculus with various settings of Doppler gain, velocity range, color filter, focal depth, and acoustic power. RESULTS: A color artifact was present within and/or distal to 29 (39%) of 75 urinary tract calculi when the old generation system was used. Use of the new generation system produced this color artifact within and/or distal to 72 (96%) of the urinary tract calculi. The intensity of the artifact was affected by the acoustic output of the equipment. CONCLUSIONS: The twinkling sign is an artifact that can not only lead to misdiagnosis of vascular flow within a structure but also help differentiate a very small stone from other small echogenic structures when later generation "all-digital" systems are used. The color sensitivity of the system and acoustic power used are important technical factors that affect the production of this artifact. PMID- 10477886 TI - Prenatal sonographic diagnosis of ectopia cordis. AB - PURPOSE: We present a small series of prenatally diagnosed cases of ectopia cordis. METHODS: Four fetuses with prenatally diagnosed ectopia cordis were sonographically evaluated and followed up. RESULTS: The fetuses were diagnosed with ectopia cordis at 9, 13, 21, and 29 weeks' menstrual age. The case diagnosed at 9 weeks is the earliest prenatal diagnosis reported, to our knowledge. The diagnoses were based on the demonstration of a fetal heart outside the thoracic cavity with Doppler waveforms typical of intracardiac flow. One fetus had isolated ectopia cordis, and 3 had other associated anolmalies. The diagnosis was postnatally confirmed in all cases. Therapeutic abortion was done in 2 cases. One infant survived, and the fourth died shortly after birth. Chromosome study was successfully performed in 2 cases and was normal in both of them. CONCLUSIONS: This small series suggests that ectopia cordis can be readily diagnosed in utero as early as the first trimester. Later in pregnancy, sonography provides important information for planning surgical correction. PMID- 10477887 TI - Sonography of the rotator cuff and biceps tendon: technique, normal anatomy, and pathology. PMID- 10477888 TI - Sonographic findings in the anterior interosseous nerve syndrome. AB - This report details previously undescribed sonographic findings in the anterior interosseous nerve syndrome. Loss of muscle bulk, increased reflectivity, reduced perfusion on Doppler sonography, and lack of active contraction of the affected muscles were observed. These findings can aid in the localization of the pathologic process and in the exclusion of tendon rupture. Dynamic observation of muscle function and Doppler changes after exercise can also help identify the muscles involved. Both sonography and MRI may be useful in the evaluation of patients with the anterior interosseous nerve syndrome and other peripheral neuropathies. PMID- 10477889 TI - Echo-enhancing sonography of a large-vessel hemangioma of the neck. AB - Because of the slow flow in the venous spaces of large-vessel hemangiomas, demonstration of color flow signals with conventional color Doppler or power Doppler sonography may be difficult. We report the case of a 22-year-old female patient with a soft tissue tumor containing multiple fluid-filled spaces in the right supraclavicular region. Gray-scale, color, and power Doppler sonography could not differentiate between cystic lymphangioma and large-vessel hemangioma. The intravenous echo-enhancing contrast agent Levovist was administered, and a significant echo-enhancing effect on color and power Doppler imaging was demonstrated in the fluid-filled spaces and lasted for about 3 minutes. Histopathologic study of the excised tumor confirmed the sonographic diagnosis of hemangioma. Levovist appears useful in depicting slow flow in a large-vessel hemangioma. PMID- 10477890 TI - Prenatal sonographic detection of nasopharyngeal teratoma. AB - We present the case of a 34-year-old pregnant woman who had an elevated maternal serum alpha-fetoprotein level and sonographic findings of a semisolid mass protruding from the fetus's oral cavity. The large, heterogeneous mass filled the oropharynx and nasopharynx. Abnormal Doppler waveforms were detected in the umbilical artery of the fetus, who died in utero. Postmortem examination revealed a nasopharyngeal teratoma. PMID- 10477891 TI - Ureteropelvic junction obstruction associated with extrarenal pelvis: A potential cause of cystic abdominal mass anterior to a normal-appearing kidney in the newborn. AB - An extrarenal pelvis is associated with the absence of central sinus echoes on sonography. However, central sinus echoes are normally inapparent in some newborns. Furthermore, true absence of a central sinus may cause calices to simulate normal renal pyramids so that the kidney appears normal in a fetus or newborn. This case illustrates the potential for an obstructed extrarenal pelvis in a fetus or newborn to distend so that it is mistaken on sonography for a cystic anterior abdominal mass unrelated to the kidney. However, the absence of central sinus echoes should suggest the diagnosis of an obstructed extrarenal pelvis if the adjacent cystic abdominal mass is positioned to obstruct an intrarenal pelvis. PMID- 10477892 TI - Sonographic detection of multiple Staphylococcus aureus hepatic microabscesses mimicking Candida abscesses. AB - We report the sonographic, CT, and clinical findings in a patient presenting with clinical sepsis and multiple Staphylococcus aureus hepatic microabscesses. Although contrast-enhanced CT has had a higher sensitivity than sonography in detecting hepatic microabscesses in some studies, this examination was negative in our patient. On sonography, numerous small hypoechoic lesions were present. Some target-like lesions had a striking similarity to Candida albicans microabscesses. The hepatic lesions were believed to be pyogenic liver microabscesses, as several blood cultures were positive for S. aureus. Following prolonged intravenous antibiotic therapy, all the hypoechoic hepatic lesions disappeared, along with the clinical and biochemical signs of sepsis. PMID- 10477893 TI - Performance of thiophilic adsorption chromatography in the purification of rat IgG2b monoclonal antibodies from serum- and protein-free culture supernatants. AB - The usefulness of thiophilic adsorption chromatography for the purification of rat IgG2b monoclonal antibodies has been evaluated. This approach has not shown specificity for immunoglobulins; therefore, to minimize potential interferences, the purification was carried out from supernatants of hybridomas grown in serum- and protein-free conditions. The protein purity of the six final antibody preparations assayed was always >/= 97%. In addition to the easiness of this procedure, which enables one-step antibody purification, the materials employed are rather inexpensive and milligram amounts of antibody can be recovered from 1 L of supernatant. Overall, the purification of rat IgG2b monoclonals under the conditions reported here offers an advantageous alternative to other more expensive and cumbersome methods. PMID- 10477894 TI - Quantitation of meiosis activating sterols in human follicular fluid using HPLC and photodiode array detection. AB - A chromatographic assay for 4,4-dimethyl-5alpha-cholesta-8,14, 24-triene-3beta-ol (FF-MAS), and its reduced species, 4, 4-dimethyl-5alpha-cholesta-8,24-triene 3beta-ol (T-MAS), has been established for analysis of human follicular fluid (huFF). The assay also quantifies lanosterol, free cholesterol and progesterone. It was established using a pool of more than 100 individual follicular fluids from women undergoing in vitro fertilization treatment. Both FF-MAS and T-MAS were found in huFF, and can be quantified with HPLC equipped with photodiode array (PDA) detection. The examination wavelength for each analyte was chosen at the absorption maximum between 200 and 300 nm. Spike-recovery experiments revealed mean recoveries of 91 +/- 7.3% for lanosterol, 103 +/- 5.1% for FF-MAS, 104 +/- 5.5% for T-MAS, 103 +/- 4.5% for free cholesterol and 85 +/- 5.1% for progesterone. The lower recovery value for progesterone was due to a sub-optimal extraction procedure for this particular analyte, as indicated by re-extraction. The minimum amounts of FF-MAS required for quantification were 4 ng/mL and 23 ng/mL for T-MAS and lanosterol. FF-MAS was assayed to approximately 1.6 microM. T MAS and lanosterol was assayed to about half of this value. No esterification of either MAS or lanosterol could be detected in huFF. Less than 10% of cholesterol was underivatized cholesterol, as more than 10 times the amount of free cholesterol could be assayed after extended saponification. This method can be used for evaluating the accumulation of MAS in huFF and its correlation to oocyte quality and fertilization parameters in in vitro fertilization programmes. PMID- 10477895 TI - Molecule imprinting chiral stationary phase. AB - Molecule imprinting polymers (MIPs) with high chiral selectivity for N(alpha) protected amino acids were synthesized in polar solvent using acrylamide (AM) or combined functional monomer methacrylic acid (MAA) + vinyl pyridine (VP). Factors that influence the chiral selectivity of MIPs and mechanisms of the chiral recognition process were investigated. It was found that the rigid structure and the polar functional group of the print molecule were very important for the chiral selectivity of MIPs. For MIPs made using a combined functional monomer, ionic and hydrophobic interaction may be responsible for the chiral recognition process in aqueous media. The number of binding sites on MIPs and dissociation constants for the complex of enantiomers and MIPs were determined by frontal chromatography analysis. PMID- 10477896 TI - Retention-activity relationship studies of benzodiazepines by micellar liquid chromatography. AB - Solute partitioning into lipid bilayers of biological membranes is the basis for drug and metabolite uptake, passive transport across membranes and bioaccumulation. In order to emulate in vitro the partitioning process in the biomembranes, different approaches have been proposed. The use of micellar solutions as mobile phases in reversed-phase liquid chromatography (micellar liquid chromatography, MLC) has proven to be valid in the prediction of the biological activities of local anesthetics, catecholamines and barbiturates. In this paper we focus our attention on benzodiazepines. The retention of benzodiazepines using different concentrations of Brij35 as micellar mobile phase in modified C(18) stationary phase was studied. Quantitative retention-activity relationships (QRAR) in MLC were investigated for these compounds. An adequate correlation between the capacity factors (log k) and the toxicity (log LD(50)) and anxiolitic activity (log ED(50)) of benzodiazepines was obtained. PMID- 10477897 TI - Determination of drug-plasma protein binding using human serum albumin chromatographic column and multiple linear regression model. AB - Reversible attachment to serum proteins plays a significant role in pharmacokinetics and pharmacodynamics, and a clear understanding of this process is fundamental in the development of the rational use of many therapeutics agents. Over the last few years, it has been demonstrated that immobilized human serum albumin (HSA) could be used to estimate plasma protein binding. A series of 40 structurally unrelated pharmaceutical compounds were chromatographed on an immobilized HSA column in order to construct a protein binding 'calibration curve' and multiple linear regression system. When studying the relationship between the chromatographic retention and the percentage of binding determined in vitro, a good correlation can be observed (r(2) = 0.799) using a wide variety of compounds with different binding affinities (from 0 to 99% binding). Using a quantitative structure-retention relationships (QSRR) approach to analysing chromatographic data, the correlation was improved compared to the traditional approach (r(2) = 0.824). PMID- 10477898 TI - Determination of glycyrrhizin in radix glycyrrhizae and its preparations by capillary zone electrophoresis. AB - A capillary zone electrophoresis method was set up for the separation and determination of glycyrrhizin in Chinese medicinal preparations. Concentrations of Na(2)B(4)O(7) were optimized, which showed that glycyrrhizin in the sample could be separated from interference in the running buffer of 30 mmol/L Na(2)B(4)O(7). Using declofenac as internal standard, the simple method was linear in the range 25-300 microg/mL of glycyrrhizin, and good reproducibility was obtained. The extracts of Radix glycyrrhizae and its preparations could be injected directly for analysis without any pretreatment. PMID- 10477899 TI - Chiral HPLC resolution of monoepoxides derived from 6,9-dienes and its application to stereochemistry assignment of fruit-piercing noctuid pheromone. AB - The resolution of insect pheromonal cis-monoepoxy racemates derived from (Z,Z) 6,9-dienes was examined employing chiral HPLC columns, and the results showed that a normal-phase column (Chiralpak AD) was suitable for both 6,7- and 9,10 epoxides with a C(17)-C(23) straight chain, as was a reversed-phase column (Chiralcel OJ-R) for the 6, 7-epoxides. To determine the absolute configuration of each separated enantiomer applying a modified Mosher's method, the epoxy ring was opened by methanolysis, and the (1)H-NMR data of (S)- and (R)-MTPA esters of the methoxyalcohols produced were analyzed. Further, the hydrogenated product of each enantiomer was chromatographed on the OJ-R column referring to the corresponding authentic chiral compounds with a saturated chain, which were prepared by a Sharpless epoxidation reaction. These analyses showed that the levorotatory 6,7- and 9,10-epoxides with shorter t(R)s possess 6S,7R and 9R,10S configuration, respectively, and the dextrorotatory enantiomers with longer t(R)s possess the opposite configuration. Utilizing this chiral HPLC, it was revealed that an abdominal tip extract of the fruit-piercing moth, Oraesia excavata Butler (Lepidoptera: Noctuidae), included (9S,10R)-(Z)-9, 10-epoxy-6-henicosene as a main sex pheromone component. The synthetic 9,10-epoxide with this configuration, which was separated from the racemate, exhibited stronger activity in electrophysiological and field tests against male moths than the enantiomer. PMID- 10477900 TI - Fluorimetric determination of D- and L-lactate derivatized with 4-(N, N dimethylaminosulfonyl)-7-piperazino-2,1,3-benzoxadiazole (DBD-PZ) by high performance liquid chromatography. AB - A highly sensitive method, based on fluorescence derivatization with 4-N,N dimethylaminosulfonyl-7-piperazino-2,1,3-benzoxadiazol e (DBD-PZ), was developed for the determination of D- and L-lactic acid. High-performance liquid chromatographic separation of the lactic acid derivative was achieved using an octadecylsilica column followed by enantiomeric separation on a phenylcarbamoylated beta-cyclodextrin chiral column. The separation factor for D, L-lactic acid derivatives was 1.30 using MeOH/H(2)O (80/20) as a mobile phase, and the detection limits were approximately 360 and 300 fmol on column for D- and L-lactic acid derivative, respectively. The proposed method was applied to determine D- and L-lactate in a wine and a lactic drink. Both D- and L-lactate could be determined simultaneously, with the precisions ranging from 3.50 to 5.78% for intra-day and 4.28-9.92% for inter-day determinations. The relative recovery was in the range of 91.6-100%. PMID- 10477901 TI - Sensitive triple-quadrupole mass spectrometric assay for the determination of BMS 181885, a 5-HT1 agonist, in human plasma following solid phase extraction. AB - A sensitive, selective, accurate, precise and reproducible triple-quadrupole liquid chromatographic-mass spectrometric assay was developed and validated for BMS-181885 (I), a 5HT1 agonist, in human plasma using BMS-181101 as the internal standard (IS). The method involved solid phase extraction of plasma containing I and the IS using Isoelute CN cartridges. The supernatant was then evaporated to dryness at 40 degrees C. The residue was dissolved in 100 microL of the injecting solvent. The HPLC column was ODS-3, 2 x 100 mm. The mobile phase comprised 10 mM ammonium formate (pH = 4) and acetonitrile, 55:45 v/v, used in an isocratic condition. The mass spectrometer was programmed to admit the protonated molecules at m/z 461 (I) and m/z 370 (IS) via the first quadrupole filter and to select reaction monitoring of ions at m/z 152 for I and IS for the quantification. Standard curves were fitted to a weighted quadratic function over the concentration range 0.2-200 ng/mL. The lowest standard concentration (0.2 ng/mL) was experimentally established as the lower limit of quantitation of the assay. The mean predicted quality control concentrations deviated within +/- 11% of the corresponding nominal values; the intra-assay and inter-assay precisions were within 7.0% relative standard deviation. I was stable in the injection solvent at 4 degrees C for at least 24 h and for at least three freeze-thaw cycles. Freezer stability of I in plasma was demonstrated for at least 3 months. The extraction recovery of I was established as 97%. The validated assay was applied to a pharmacokinetic study of I in humans. PMID- 10477902 TI - Determination of melatonin in pharmaceutical formulations and human plasma by gas chromatography-electron impact mass spectrometry. AB - A simple and reliable method for gas chromatography-mass spectrometry (GC-MS) has been developed for the determination of melatonin in commercially available tablet formulations and human plasma. The dissolution of the tablets in ethyl acetate was simply obtained by sonication. The filtrate of the resultant solution, after concentration, was derivatized with pentafluoropropionic anhydride (PFPA) and analysed by GC-MS using splitless injection. The linear response range for melatonin was 5-250 ng on column. The same method can be used, with minor modifications, for the determination of melatonin in spiked human plasma in the range 50-1000 pg/mL plasma. PMID- 10477903 TI - Measurements of free and total PSA, tissue polypeptide-specific antigen (TPS), and CYFRA 21-1 in prostate cancer patients under intermittent androgen suppression therapy. AB - BACKGROUND: The present study evaluated monthly measurements of free and total prostate-specific antigen (PSA), and the tumor proliferation markers tissue polypeptide-specific antigen (TPS) and cytokeratin fragment 21-1 (CYFRA 21-1) in patients with advanced prostate cancer receiving intermittent androgen suppression therapy (IAS). METHODS: Thirty-four men received alternating cycles of 8 month androgen suppression and treatment cessation (mean duration, 10.3 months) until PSA increased to >20 microg/l. Measurements of testosterone, percentage of free PSA, TPS, and CYFRA 21-1 were performed using ELISA and RIA assays. RESULTS: Periods of androgen suppression resulted in reversible reductions of testosterone (from 6 +/- 0.8 to <0.58 ng/ml), PSA (from 31.2 +/- 4.5 to <1.7 microg/l), and prostatic volume (mean reduction, 22.2 +/- 4.6%), indicating apoptotic regression of the tumors. Upon treatment cessation, testosterone increased to 6.1 +/- 0.56 ng/ml within 2 months, followed by an increase of PSA to 5.8 +/- 0.8 microg/l. The mean percentage of free PSA (15.1 +/ 2.6%) exhibited no significant change during the whole IAS cycle. TPS showed a decrease of 50% after 3 months, and CYFRA 21-1 a 25% decrease after 7 months of androgen suppression treatment. During treatment cessation, TPS exceeded the normal cutoff value of 90 U/l late in tumor regrowth (9-11 months), whereas CYFRA 21-1 remained below the normal cutoff value of 3.3 ng/ml. CONCLUSIONS: PSA is the best and most sensitive marker of prostate cancer regression and regrowth during IAS cycles of the markers tested in this study. Free PSA constitutes approximately 15% of total PSA (range, 5-32%), and its percentage showed no significant change during IAS cycles. The TPS and CYFRA 21-1 proliferation marker changes in IAS seem to be related mainly to effects on normal androgen-dependent tissues. PMID- 10477904 TI - CXC-chemokines stimulate invasion and chemotaxis in prostate carcinoma cells through the CXCR2 receptor. AB - BACKGROUND: Metastasis of prostate carcinoma requires invasion through the basement membrane, a thin extracellular matrix that underlies the epithelial cells, which must be breached by tumor cells invading into surrounding tissue. The CXC-chemokines, which have been shown to promote the migration of neutrophils and carcinoma cells, are candidates to influence prostate carcinoma-cell invasion. METHODS: CXC-chemokines were examined for the ability to stimulate prostate cell line PC3 invasion in vitro through a reconstituted basement membrane and long-term migration and short-term adhesion to laminin, a major component of the basement membrane. RESULTS: PC3 cells responded to IL-8 and GROalpha with a 1. 6-2-fold increase in invasion through reconstituted basement membrane. A corresponding 2-3-fold increase in chemotaxis toward IL-8 and GROa was seen on laminin. Anti-CXCR2 antibody inhibited IL-8-stimulated migration. Expression levels of the beta(1) integrins were not changed by IL-8, and alpha(6beta1) integrin was used for both stimulated and baseline migration. In addition to the increases in migration and invasion, 2-6-fold transient increases in adhesion on laminin were seen with both IL-8 and GROalpha. CONCLUSIONS: These results suggest that the CXC-chemokines stimulate migration and invasion in part by altering the activation state of the beta(1) integrins. The CXC-chemokines act on prostate carcinoma cells through the CXCR2 receptor to promote behavior important for metastasis, and as such may be important in prostate carcinoma progression and metastasis. PMID- 10477905 TI - Comparison of urological irrigating fluids containing glycine and mannitol in volunteers. AB - BACKGROUND: We compared symptoms and indices of fluid distribution after experimental administration of glycine and mannitol solutions, since these irrigating fluids are sometimes absorbed by the patient during genitourinary surgery. METHODS: Glycine 1.5% and mannitol 3%, both with ethanol 1% added, were given by intravenous infusion at a rate of 0.5 ml/kg/min during 30 min to 12 male volunteers. Symptoms, cognitive status, hemodynamics, electrocardiogram during 24 hr, computerized tomography of the brain, bioimpedance, blood chemistry, and breath ethanol concentrations were recorded. RESULTS: Glycine was associated with more symptoms than mannitol (P< 0.006), but the cognitive status, computerized tomography examinations, electrocardiograms, and breath ethanol concentrations did not differ between the solutions. The urinary excretion of fluid and sodium was greater after mannitol (P< 0.04), while only the glycine infusions hydrated the cells (P< 0.05). For both fluids, the intravascular and interstitial volumes were below baseline 3 hr after the experiment started (P< 0.01). CONCLUSIONS: Glycine 1.5% had a higher tendency than mannitol 3% to cause symptoms and to accumulate in the cells. PMID- 10477906 TI - In situ hybridization study of PSP94 (prostatic secretory protein of 94 amino acids) expression in human prostates. AB - BACKGROUND: The prostatic secretory protein of 94 amino acids (PSP94), also named beta-microseminoprotein, is one of the major proteins secreted by the human prostate. However, its value as a prognostic marker for prostate cancers is still under debate. The aim of the present study was to examine the expression pattern of this protein in fetal, pubertal, and aged human prostates. METHODS: Nonisotopic in situ hybridization using a digoxigenin-labeled riboprobe for PSP94 and immunohistochemistry were used to demonstrate the expression of PSP94 in different regions or zones of fetal, pubertal, and adult human prostates. Its localization pattern was also compared with those of two other major secretory proteins, prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP), by immunohistochemistry. RESULTS: PSP94 mRNA and its protein were localized to the secretory epithelium of normal pubertal and adult human prostates. No hybridization signal and immunoreactivity of PSP94 were seen in fetal prostates at 6-7 months of gestation, whereas some glandular cells were positive to PSA and PAP immunostainings. In the adult prostates, PSP94 expression was intense in the acini in the peripheral zone, less intense in the transition zone, and variable in the central zone. Such a zonal expression pattern was more apparent in the pubertal prostates. However, no obvious differential expression pattern was observed in the immunohistochemistry of PAP and PSA, which showed a uniform staining of the secretory epithelia of the acini in all anatomic zones. The hybridization signals and immunoreactivity of PSP94 became reduced or lost in premalignant prostatic intraepithelial neoplastic lesions and different grades of prostatic carcinomas. CONCLUSIONS: Fetal prostates at 6-7 months of gestation already synthesize PSA and PAP but not PSP94. The delayed expression of PSP94 appears to correlate with the development of the prostate gland. A differential expression pattern of PSP94 is demonstrated in different anatomical zones, showing that this protein is more expressed and synthesized in the acini in the peripheral zone than in the central and transition zones. However, such a zonal pattern is not seen in the immunohistochemistry of PSA and PAP. The present study also shows that PSP94 is downregulated in different grades of prostate cancers. PMID- 10477907 TI - Regulation of prostatic stromal cell growth and function by transforming growth factor beta (TGFbeta). AB - BACKGROUND: The majority of the overgrowth in benign prostatic hyperplasia (BPH) specimens is comprised of connective tissue. Factors that control stromal growth in the prostate are poorly understood; however, members of the transforming growth factor beta (TGFbeta) family may be of particular importance in the etiology of BPH. METHODS: Thirty-two low-passage stromal cultures were generated from human prostatectomy specimens. Their stromal origin was confirmed and expression of TGFbetas analyzed by duplex reverse transcription-polymerase chain reaction (RT-PCR). Challenge experiments were designed to study the effects of exogenous TGFbeta1 on stromal cell growth and synthesis of extracellular matrix components. RESULTS: The expression of TGFbetas 1, 2, and 3 was demonstrated in all 32 cell strains. The stromal origin of the cell lines was confirmed. Exogenous TGFbeta1 added to stromal cultures resulted in inhibition of cell growth and increased production of type I collagen. CONCLUSIONS: The prostatic stromal cell strains we have developed are a reliable mod- el for investigating prostatic connective tissue biology. The challenge experiments with TGFbeta1 provide further evidence for the involvement of TGFbetas in prostatic enlargement, as modulators of the extracellular matrix in the absence of growth stimulation. PMID- 10477908 TI - "Decoy" of androgen-responsive element induces apoptosis in LNCaP cells. AB - BACKGROUND: In an androgen-dependent manner, the androgen receptor (AR) binds to the androgen-responsive element (ARE) in the regulatory region of target genes. We hypothesize that an "ARE decoy, " a double-stranded oligonucleotide containing the same DNA sequence as ARE, can inhibit prostatic proliferation by competitive inhibition of AR transcriptional activity. METHODS: We synthesized a 23-mer ARE decoy based on the deduced ARE sequence at the promoter region of the human prostate-specific antigen (PSA) gene. The nuclear extract was prepared from LNCaP cells, and DNA-protein interactions were examined by gel shift assay. Then the antiandrogen effect of the ARE decoy was studied in LNCaP cells transfected with the ARE decoy by lipofection. After 24-hr incubation with 10(-9) M dihydrotestosterone (DHT), induction of apoptosis was examined by DNA fragmentation. RESULTS: The gel shift assay demonstrated specific binding of the ARE decoy to the LNCaP nuclear protein which is most likely AR. The transfection experiment showed DNA fragmentation in the ARE decoy-transfected cells despite the presence of DHT, though not in the cells transfected with the control decoy. CONCLUSIONS: The ARE decoy had an antiandrogen effect and induced apoptosis in LNCaP cells. This ARE decoy may become a potential therapeutic tool for prostate cancers when combined with a highly efficient transfection method. PMID- 10477909 TI - Circulating levels of interleukin-6 in patients with hormone refractory prostate cancer. AB - BACKGROUND: Interleukin-6 (IL-6) is a cytokine that plays a central role in host defense due to its wide range of immune and hematopoietic activities. It is found in high levels in human ejaculate, and has recently been found to regulate prostate-specific protein expression in prostate cancer cells through nonsteroidal activation of the androgen receptor. IL-6 may be a candidate mediator of morbidity in patients with metastatic disease. We attempted to evaluate the potential of circulating IL-6 levels as a marker of disease progression. MATERIALS AND METHODS Serum IL-6, prostate specific antigen (PSA), percent free PSA (%fPSA), and prostate-specific membrane antigen (PSMA) were measured using commercially available assays in 407 men, including 15 controls. The rest of the study population had clinical or histologic evidence of prostate diseases, including 41 patients with chronic prostatitis, 167 with benign prostatic hyperplasia (BPH), 8 with high-grade prostatic intraepithelial neoplasia (PIN), 88 with localized prostate cancer, 22 with local recurrence after treatment of primary tumor, 4 with advanced untreated disease (nodal or bony metastases), 23 with advanced hormone dependent disease, and 39 with advanced hormone refractory disease (PSA > 1.0 ng/ml while on hormone treatment and/or evidence of disease progression). None had history of concurrent malignancy or acute inflammatory condition. Kruskal-Wallis analysis of variance and Spearman's correlation analysis were used for statistical analyses. RESULTS: Serum levels of IL-6 were significantly elevated in patients with clinically evident hormone refractory disease (5.7 +/- 1.9 pg/ml) and statistical significance was seen when comparing the elevated serum IL-6 levels to those in normal controls, prostatitis, BPH, and localized and recurrent disease, (P values < 0.01). Compared to serum levels of controls and BPH, PSA was significantly elevated in advanced untreated disease and hormone refractory groups (P < 0.05). Percent fPSA was significantly lower in all cancer patients but the hormone refractory. Serum PSMA was elevated in advanced untreated prostate cancer. Serum IL-6 showed positive correlation with PSMA and negative correlation with serum PSA but did not attain statistical significance. CONCLUSIONS: Serum IL-6 levels are significantly elevated in hormone-refractory prostate cancer patients and may be a surrogate marker of the androgen independent phenotype. PMID- 10477910 TI - Use of a yeast assay to detect functional alterations in p53 in prostate cancer: review and future directions. AB - BACKGROUND: While many studies have suggested that p53 mutations are common in human cancers, the functional activity of these mutant alleles has not yet been fully addressed. We believe that information about the functional status of individual p53 mutants will prove to be important for a better understanding of the role of p53 in tumor development and progression. Ultimately, this information could also influence treatment decisions for individual cancer patients. METHODS: A recently developed yeast functional assay can be used to assess the transactivational activity of p53 mutants. Furthermore, this assay is more sensitive than single strand conformational polymorphism (SSCP) for detection of p53 mutations. In this review, we summarize the mechanism of this new technique and describe its applications in cancer research, with an emphasis on prostate cancer. RESULTS: The use of the yeast functional assay provides a simple, sensitive, and reproducible method for detecting p53 mutations and for determining the transactivational activity and dominant-negative role of individual p53 mutants. CONCLUSIONS: This method may be adapted to analyze other transcriptional factors, including the human androgen receptor. PMID- 10477911 TI - Biologically based analysis of the data for the Colorado uranium miners cohort: age, dose and dose-rate effects. AB - This study is a comprehensive analysis of the latest follow-up of the Colorado uranium miners cohort using the two-stage clonal expansion model with particular emphasis on effects related to age and exposure. The model provides a framework in which the hazard function for lung cancer mortality incorporates detailed information on exposure to radon and radon progeny from hard rock and uranium mining together with information on cigarette smoking. Even though the effect of smoking on lung cancer risk is explicitly modeled, a significant birth cohort effect is found which shows a linear increase in the baseline lung cancer risk with birth year of the miners in the cohort. The analysis based on the two-stage clonal expansion model suggests that exposure to radon affects both the rate of initiation of intermediate cells in the pathway to cancer and the rate of proliferation of intermediate cells. However, in contrast to the promotional effect of radon, which is highly significant, the effect of radon on the rate of initiation is found to be not significant. The model is also used to study the inverse dose-rate effect. This effect is evident for radon exposures typical for mines but is predicted to be attenuated, and for longer exposures even reversed, for the more protracted and lower radon exposures in homes. The model also predicts the drop in risk with time after exposure ceases. For residential exposures, lung cancer risks are compared with the estimates from the BEIR VI report. While the risk estimates are in agreement with those derived from residential studies, they are about two- to fourfold lower than those reported in the BEIR VI report. PMID- 10477912 TI - Characteristics of the cohort of workers at the Mayak nuclear complex. AB - At Branch No. 1 of the Russian State Research Center "Biophysics Institute", a registry has been created of workers at the "Mayak" Production Association, the first nuclear complex in Russia. This registry includes 18,830 persons hired at Mayak's nuclear reactors and radiochemical and plutonium production plant between 1948 and 1972. Twenty-five percent of these workers are women. As of December 31, 1994, the vital status is known for approximately 90% of the cohort members. A total of 5,118 persons have died. The cause for 97% of total deaths has been ascertained. The cohort members were exposed to both external gamma radiation and internal radiation from incorporated plutonium. The plutonium body burden has been measured in 30% of the cohort members with potential for plutonium exposure. External gamma-ray doses were in the range from tenths of milligrays to about 10 Gy, and plutonium body burdens were up to about 260 kBq. In view of the nature of the Mayak worker cohort, it has the potential to provide reasonably precise, quantitative estimates of the long-term health effects associated with chronic low-dose-rate exposure to external gamma radiation as well as internal radiation from plutonium. However, a number of issues must be addressed before credible risk estimates can be obtained from this cohort. These issues include the development of an appropriate internal comparison group and/or external rates and separating of the effects of internal and external exposures on risk estimates. PMID- 10477913 TI - Effects of radiation on incidence of primary liver cancer among atomic bomb survivors. AB - We describe the radiation risk for primary liver cancers between 1958 and 1987 in a cohort of atomic bomb survivors in Hiroshima and Nagasaki, Japan. The analysis is based on a comprehensive pathology review of known or suspected liver neoplasms that generated 518 incident, first primary cases, mostly hepatocellular carcinoma. Excess relative risk from atomic bomb radiation was linear: 0.81 per sievert weighted liver dose (95% CI [0.32, 1.43]; P < 0.001). Males and females had similar relative risk so that, given a threefold higher background incidence in males, the radiation-related excess incidence was substantially higher in males. Excess risk peaked for those with age at exposure in the early 20s; there was essentially no excess risk in those exposed before age 10 or after age 45. Whether this was due to a difference in sensitivity or possible confounding by other factors could not be addressed retrospectively in the full cohort. A paucity of cholangiocarcinoma and hemangiosarcoma cases suggested that they are not significantly associated with whole-body radiation exposure, as they are with the internal alpha-particle-emitting radiological contrast medium Thorotrast. Because most of the radiation-related excess cases occurred among males, it is important to ascertain what factors put men at greater risk of radiation-related liver cancer. PMID- 10477914 TI - Studies of the mortality of atomic bomb survivors. Report 12, part II. Noncancer mortality: 1950-1990. AB - This report updates the data on noncancer mortality for 86,572 atomic bomb survivors with dose estimates in the Radiation Effects Research Foundation's Life Span Study cohort. The primary analyses are based on more than 27,000 noncancer disease deaths that occurred in the cohort between October 1, 1950, and December 31, 1990, 30% more than in the previous report. The present analyses strengthen earlier findings of a statistically significant increase in noncancer disease death rates with radiation dose. Increasing trends are observed for diseases of the circulatory, digestive and respiratory systems. Rates for those exposed to 1 Sv are elevated about 10%, a relative increase that is considerably smaller than that for cancer. However, estimates of the number of radiation-related noncancer deaths in the cohort to date (140 to 280) are 50 to 100% of the number for solid cancer. The data do not yet clarify the shape of the dose response. There is no significant evidence against linearity, but the data are statistically consistent with curvilinear dose-response functions that posit essentially zero risk for doses below 0.5 Sv. Similarly, while the data are consistent with substantial variation in the excess relative risk with age at exposure or attained age, there is no statistically significant dependence on these factors. In view of the small relative risks and the lack of understanding of biological mechanisms, we emphasize consideration of whether the findings could be explained by misclassification, confounding or selection effects. Based on available data, we conclude that such factors are unlikely to fully explain the observed dose response. A significant dose response is also seen for deaths from blood diseases with an excess relative risk that is several times greater than that seen for solid cancer. Particular attention is paid to the possibility that this apparent effect is a consequence of the attribution of leukemia or other cancer deaths to noncancer blood diseases. We find that misclassification does not explain this excess risk. As in earlier reports, suicide rates tend to decrease with increasing dose. PMID- 10477915 TI - Preferential liver irradiation enhances hematopoiesis through a thrombopoietin independent mechanism. AB - Liver synthesizes thrombopoietin, which is a major cytokine involved in the production of hematopoietic cells. The purpose of this study was to examine the effects of preferential liver irradiation on expression of thrombopoietin and production of hematopoietic cells. About 70% of the liver of C57BL6/J mice was irradiated with 20 Gy of gamma rays. Exposure to ionizing radiation enhanced hematopoietic progenitors and megakaryocyte frequency in bone marrow and induced a transient increase in platelet and neutrophil counts that peaked 14 days after irradiation. The concentration of thrombopoietin was increased in serum as early as 5 h after liver irradiation and was still elevated at day 14. By using Northern blot analysis and an RNase protection assay, we showed that thrombopoietin mRNA was increased in the irradiated liver. To determine whether thrombopoietin was involved in the stimulation of hematopoiesis, we irradiated mice in which thrombopoietin deficiency had been induced by homologous recombination. Platelet levels were increased in both heterozygous and homozygous thrombopoietin-deficient mice with a magnitude similar to that obtained in normal mice. In summary, our data demonstrate that local irradiation of the abdomen encompassing the liver leads to stimulation of hematopoiesis through a thrombopoietin-independent mechanism. PMID- 10477916 TI - In vivo and in vitro radioprotective effects of the prostaglandin E1 analogue misoprostol in DNA repair-proficient and -deficient rodent cell systems. AB - The radioprotective effect of a stable prostaglandin E(1) analogue, misoprostol, was studied in cells from mice with severe combined immunodeficiency (SCID) and in normal cells using X-ray-induced chromosomal aberrations and/or cell killing as the end points. The results clearly show misoprostol-induced radioprotective effects in spermatocytes of the first meiotic division when analyzed for X-ray induced chromosomal aberrations. The protective effect was independent of Trp53 (formerly known as p53) status. Since spermatocytes are relatively easy to isolate, this appears to be a suitable in vivo model that will allow biochemical studies of the mechanisms involved in radioprotection mediated by misoprostol. Using transfected CHO-K1 cells that stably express a PGE(2) receptor (CPE cells), significant radioprotection mediated by misoprostol from both chromosome breakage and cell death could be demonstrated under in vitro conditions. In addition, evidence was obtained indicating that the degree of radioprotection was dependent on the cell cycle and that S-phase cells were less responsive to misoprostol mediated radioprotection. These results suggest that CPE cells may be a suitable in vitro model for further studies on the cellular pathways involved in radioprotection by misoprostol in particular and prostaglandins in general. PMID- 10477917 TI - The farnesyltransferase inhibitor FTI-277 suppresses the 24-kDa FGF2-induced radioresistance in HeLa cells expressing wild-type RAS. AB - In this paper, we describe the effect of the inhibitor of farnesyltransferase (FTI-277) on radioresistance induced by the 24-kDa isoform of FGF2 in human cells expressing wild-type RAS. Treatment with FTI-277 (20 microM) for 48 h prior to irradiation led to a significant decrease in survival of radioresistant cells expressing the 24-kDa isoform (HeLa 3A) but had no effect on the survival of control cells (HeLa PINA). The radiosensitizing effect of FTI-277 is accompanied by a stimulation of postmitotic cell death in HeLa 3A cells and by a reduction in G(2)/M-phase arrest in both cell types. These results clearly demonstrate that at least one farnesylated protein is involved in the regulation of the radioresistance induced by the 24-kDa isoform of FGF2. Furthermore, the radiation induced G(2)/M-phase arrest is also under the control of farnesylated protein. This work also demonstrates that FTase inhibitors may be effective radiosensitizers of certain human tumors with wild-type RAS. PMID- 10477918 TI - Chromosome aberrations induced in human lymphocytes by 3.45 MeV alpha particles analyzed by premature chromosome condensation. AB - Premature chromosome condensation (PCC) experiments using human lymphocytes with centromere staining have shown that after exposure to 3.45 MeV alpha-particle radiation, the full number of dicentric chromosomes appears when the cell fusion protocol is applied immediately after irradiation. In this case, the time available for repair and misrepair of DNA damage is only about 30 min. The number of dicentrics does not change with a further increase in the time available for chromatin rearrangement. This fast response confirms the expectation based on our previous experiments using PCC with 150 kV X rays in which the alpha component of the yield of dicentrics was found to appear when the cell fusion protocol was applied immediately after irradiation, whereas the beta component was delayed by several hours. The time constant for rejoining of the excess acentric chromosome fragments is found to be donor-specific and not to differ for alpha particles and X rays, but alpha-particle radiation leaves a larger fraction of the excess acentric fragments unrejoined. The RBEs of the 3.45 MeV alpha-particle radiation compared to 150 kV X rays, evaluated for the alpha component for the yield of dicentrics and for the yield of unrepaired acentric fragments, have almost equal values of about 4. This is consistent with data in the literature on chromosome aberrations observed in metaphase that show the equality of the RBE values for production of dicentrics and acentric fragments. Our experimental results concerning the fast kinetics of the alpha component of the yield of exchange-type chromosome aberrations are not consistent with Lea's pairwise lesion interaction model, and they support the proposed alternative mechanism of lesion-nonlesion interaction between chromatin regions carrying clustered DNA damage and intact chromatin regions. PMID- 10477919 TI - The complexity of radiation-induced DNA damage as revealed by exposure to cell extracts. AB - The rejoining of single-strand breaks (SSBs) induced in plasmid DNA in the presence of 10 mmol dm(-3) Tris scavenger by aluminum K (Al(K)) ultrasoft X rays has been compared with that for SSBs induced by gamma radiation. The Al(K) ultrasoft X rays interact to produce low-energy secondary electrons, which are thought to be the main contributors to the formation of complex damage by low-LET radiations. The rejoining of radiation-induced SSBs was investigated using human whole cell extracts. The efficiency of rejoining of SSBs induced by Al(K) ultrasoft X rays is less than that observed for gamma-ray-induced SSBs. From the similarity of the extent of rejoining of SSBs induced by gamma rays under aerobic and anaerobic conditions, the chemical nature of the stand break termini does not significantly influence SSB rejoining. A simple nick induced in plasmid DNA by gpII protein is rejoined rapidly compared with the slower rejoining processes for radiation-induced SSBs. Therefore, ligation is not rate-determining in processing radiation-induced SSBs. This study provides further evidence that nonrejoining of radiation-induced SSBs reflects the complexity of DNA damage. From comparison of the extent of rejoining of SSBs induced by different radiations, it is inferred that double-strand breaks represent only a minor component of the overall yield of complex damage. PMID- 10477920 TI - Photodynamic properties of meta-tetra(hydroxyphenyl)chlorin in human tumor cells. AB - The photodynamic properties of a second-generation photodynamic sensitizer, meta tetra(hydroxyphenyl)chlorin (mTHPC) were studied by dye-sensitized photoinactivation (650 nm) of HT29 human adenocarcinoma cells in culture. The photocytotoxicity of mTHPC in vitro depended on the presence of molecular oxygen. A strong inhibition of the photocytotoxicity of mTHPC was observed upon addition of sodium azide, a known singlet oxygen quencher. Photocytotoxicity was not inhibited by scavengers of superoxide anion radical, hydrogen peroxide and hydroxyl radicals. We suggest that mTHPC photosensitizes cell killing predominantly by type II, singlet oxygen-mediated photodynamic reactions. Illumination of cells preloaded with mTHPC induced peroxidation of membrane lipids. Inhibition of photoperoxidation by alpha-tocopherol (0.1 mM) present during illumination did not result in any decrease in toxicity, suggesting that reactions of lipid peroxidation play only a minor role in the overall photocytotoxic effect of mTHPC. PMID- 10477921 TI - Lymphocyte subset analyses in blood, spleen and lymph nodes of female Sprague Dawley rats after short or prolonged exposure to a 50 Hz 100-microT magnetic field. AB - Based primarily on the results of in vitro studies, it has been suggested that power-line (50 or 60 Hz) magnetic fields (MFs) may reduce immune function, which could lower resistance to infection or cancer. This study was conducted to evaluate the influence of acute and chronic in vivo exposure to a linearly polarized 50 Hz MF on immune function in female Sprague-Dawley rats. Groups of rats were exposed continuously to the MF at a flux density of 100 microT for periods of 3 days, 14 days or 13 weeks. For each exposure period, one control group of rats was sham-exposed together with each MF-exposed group. Experimental end points included analyses of T-lymphocyte subsets as well as other immune cells involved in cell-mediated immune responses, i.e. natural killer (NK) cells, B lymphocytes, macrophages, and granulocytes in blood, spleen and mesenteric lymph nodes. In addition, immunohistochemical methods were used to detect proliferating and apoptotic cells in the various compartments of spleen tissue. The results obtained failed to demonstrate a significant effect of short or prolonged MF exposure on different types of leukocytes, including lymphocyte subsets. Furthermore, the experiments on the in vivo proliferation activity of lymphocytes and the extent of apoptosis in spleen samples did not indicate a difference between the MF-exposed and sham-exposed groups, indicating that MF exposure does not affect the mechanisms involved in the control of lymphocyte homeostasis. The lack of MF effects in the immune tests used in the present in vivo study makes it highly unlikely that MF exposure induces immunotoxicity, at least under the experimental conditions used. However, the data do not exclude the possibility that functional alterations in T-cell responses to mitogens and in NK cell activity as recently described for MF-exposed rodents may be one mechanism involved in the carcinogenic effects of MF exposure observed in some models of co-carcinogenesis. PMID- 10477922 TI - Repair of radiation-induced DNA strand breaks does not occur preferentially in transcriptionally active DNA. AB - Certain DNA base lesions induced by ionizing radiation or oxidative stress are repaired faster from the transcribed strand of active genes compared to the genome overall. In this study, it was investigated whether radiation-induced DNA strand breaks are preferentially repaired in active genes compared to the genome as a whole in CHO cells. The alkaline unwinding technique coupled to slot-blot hybridization with specific DNA probes was used to study the induction and repair of DNA strand breaks in defined DNA sequences. Results using this technique showed a linear dose response for the formation of radiation-induced DNA strand breaks in the dihydrofolate reductase (DHFR) gene. Furthermore, the half-life of radiation-induced strand breaks was less than 5 min in the DHFR gene, in the ribosomal genes, and in the genome as a whole. These results suggest that the repair of DNA strand breaks is fast and uniform in the genome of mammalian cells. PMID- 10477923 TI - Towards a psychological model of midlife crisis. AB - Midlife crisis in men is seen as a process of intensive and subjectively difficult transition of the self dealing with a reinterpretation of time perspective, the confrontation with death as a future personal event, the re evaluation of life values and goals, and planning the second half of life. Midlife crisis arises on the relationships between the changing sociopsychological situation and internal predispositions. This study was conducted in Poland, using a sample of 144 men (aged 35-45 years). Measures were the Midlife Crisis Questionnaire, the Time Orientation Scale, the Adjective Check List, the modified version of the Ways of Coping Checklist, and the Value crisis Questionnaire. The findings indicated that the midlife crisis consists of three relatively independent dimensions, extracted by factor analysis, namely, (i) intensity of symptoms focused on changes in the self-concept, (ii) psychological maturity, and (iii) acceptance of time passing and death. Necessary and sufficient conditions of the crisis appeared to be (1) value crisis, understood as difficulties in hierarchization, integration, and realization of values, (2) emotion-focused coping versus problem-focused coping, (3) past versus future time orientation and lack of goals for the future, (4) sense of time pressure, (5) some conscientiousness, introversion, and openness to experience. PMID- 10477924 TI - Motivation and future temporal orientation: a test of the self-handicapping hypothesis. AB - Self-handicapping motivation refers to the likelihood a person will project personal ambition into the future, make a pessimistic judgement, and then mobilise effort in the present to avoid an anticipated negative outcome. It should, therefore, be a correlate of future time perspective. This study showed for a sample of 120 first-year students that, whilst future time perspective did strongly predict scores on a measure of self-handicapping motivation, neither variable was a useful predictor of outcome. PMID- 10477925 TI - An attribution-emotion model of helping behavior. AB - To investigate the help-giving judgment of the attribution-emotion model further, data from five manifest variables of controllability, responsibility, anger, sympathy, and help-giving, were subjected to path analysis, which provided an easy interpretation of the model. Participants were 90 men and 81 women in an introductory psychology course. PMID- 10477926 TI - Why adolescent boys dream of becoming professional athletes. AB - A review of studies which investigated drawings of the ideal man and the occupational aspirations of boys (ages 11-18 years) from several countries indicated that becoming a professional athlete was a popular but unrealistic aspiration for many adolescent boys. Boys who were athletes and nonathletes from diverse ethnic groups and nationalities dreamed of becoming professional athletes. In two additional studies in the United States of America, adolescents were asked why they thought boys most often selected professional athlete as a possible future occupation. Adolescents perceived professional athletes as rich, famous, and glorified. Enhancement of status and financial gain were ranked as more important than the desire to play sports. PMID- 10477927 TI - The misery index and suicide. AB - From 1958-1992, the original misery index predicted the American suicide rate better than did a revised misery index. PMID- 10477928 TI - Mental health services in the county jail: a critical partnership? AB - This study was designed to provide a description of individuals incarcerated in a county jail and referred for mental health services. A standardized intake form was completed for 598 inmates who had contact with the mental health counselor. Analysis of the mental health status of inmates suggests that the presence of a counselor in the jail may serve an important function. Specifically, inmates referred to the counselor were not in acute distress. This suggests they may be better served by an on-site counselor rather than through the traditional method of being transported to the hospital emergency room or community mental health center for evaluation. PMID- 10477929 TI - Urban traffic noise and self-reported health. AB - This paper analyzed urban traffic noise effects on health at two different levels: intense noise and moderate noise. 42 residents of the area responded to questions on perceived noise and psychological and behavior disturbance before and after acoustic insulation was constructed. Analysis of self-reports indicated that perceived noise was associated with lower health. Also these perceived effects on health did not decrease after the acoustic isolation work was finished. PMID- 10477930 TI - Comment on the effectiveness of treatment for dissociative identity disorder. AB - Methodological criticisms of a recent study on treatment outcome of dissociative identity disorder are accurate: however, the outcome study is more methodologically sound than any previous work and therefore advances the field. The design of other studies and the conclusions to be drawn from the study under discussion are reviewed. PMID- 10477931 TI - Re-examination of assertiveness and aggressiveness as potential moderators of verbal intentions. AB - This research reports the results of a replication of Swanson and McIntyre's 1998 study of assertiveness and aggressiveness as potential moderators of verbal intentions. A convenience sample of 119 women and 71 men participated. Nonassertive subjects were least likely, while Resort-to-Aggressive subjects were the most likely, to talk with acquaintances and others. Nonassertive and Assertive subjects were significantly more likely than Aggressive ones to make recommendations. Finally, men and women scored equally assertive, but men were more likely to score aggressive. The findings generally support the earlier work. PMID- 10477932 TI - Human-dog interactions in a guide-dog training program. AB - We analyzed dyadic interactions between 12 neutered dogs (6 females and 6 males) and 44 humans (20 women, 14 men, and 10 girls) who were unfamiliar with each other. We also examined the effect of sex differences in dogs and humans as well as age differences in humans on human-dog interactions in a guide-dog training program. Female dogs more actively regulated their distance from humans than male dogs. Dogs made contact with women more frequently than with men, and men made contact with dogs more frequently than women. Girls initiated interactions with dogs more frequently than women; girls formed reciprocal interactions with dogs less frequently than women. PMID- 10477933 TI - Optimism and manic-depressive tendencies. AB - Scores on an optimism scale were negatively associated with both music and depressive tendencies in a sample of 65 college students. PMID- 10477934 TI - Physiological and psychological effects of active-alert hypnosis. AB - 10 female physical education students were given posthypnotic suggestions alluding to economy of effort in exercise. These suggestions were affirmed under self-hypnosis over two weeks. Experimental subjects pedalled on an exercise bicycle for 20 min. at 60% of their maximal work capacity while listening to suggestions from a hypnotist. The Control group (n = 9) showed no differences between mean pretest and posttest scores on physical self-efficacy, trait anxiety, state anxiety, heart rate, blood pressure, and perceived rate of exertion. The average systolic blood pressure of the experimental group was the only positive significant difference between the pretest and posttest scores. PMID- 10477935 TI - The Junior Temperament and Character Inventory: preliminary validation of a child self-report measure. AB - A preliminary effort to validate the Junior Temperament and Character Inventory with a convenience sample of 322 children ages 9 to 12 years is described. PMID- 10477936 TI - Effect of changing alcohol laws in Iceland on suicide rates. AB - After the legalization of strong beer in 1989 in Iceland, the decrease in consumption of spirits was accompanied by a decrease in the suicide rate. PMID- 10477937 TI - Depressive symptomatology and specificity of social support. AB - Psychosocial interventions should be based on research which reflects the multidimensionality of psychosocial constructs. This study investigated six categories of social support which were associated with depressive symptomatology in a sample of 531 college students. The Social Provisions Scale was used to measure social support while the Center for Epidemiological Studies-Depression Scale was used to measure depressive symptomatology. Standard multiple regression analysis was used to analyze the variance contributed by each of the six categories of social support found on the Social Provisions Scale to depressive symptomatology. Specifically, the following categories of social support were significantly associated with depressive symptomatology: Reassurance of Worth (for male and female respondents) and Attachment (for females only). Implications for practice and research are discussed. PMID- 10477938 TI - A cross-cultural comparison of attitudes toward persons with disabilities: Greeks and Greek-Americans. AB - The present study compares the attitudes of 101 Greeks and 98 Greek-Americans toward persons with disabilities. The Attitudes Toward Disabled Persons Scale was used to assess subjects' attitudes. Religiosity, education, and amount and type of contact with persons with disabilities were also measured. Analyses indicated that ethnicity accounted for a significant portion (28%) of the variance, with more positive attitudes among Greek-Americans. Also, opportunity to work with persons with disabilities accounted for 3% of the variance. The other variables did not significantly affect attitudes. PMID- 10477939 TI - Public displays of affection among Asian and Latino heterosexual couples. AB - The present study used naturalistic observation to investigate whether public touching behavior differs as a function of the interactants' race or ethnicity. Heterosexual dyads (22 Asian, 22 Latino; average age = 24 years) were observed while walking through the campus of a large western university. Some support was found for the notion that interactants from "contact" cultures, e.g., Latin America, are more contact-prone than those from "noncontact" cultures, e.g., Asia. Specifically, Latino male-female dyads were more likely to embrace while walking in public than Asian male-female dyads. PMID- 10477940 TI - Effects of humor on test anxiety and performance. AB - The present study evaluated the moderating effects of humor in test items on the hypothesized relationship between test anxiety and performance. Subjects initially completed anxiety scales, as well as coping-humor and sense-of-humor scales. 34 women and 26 men received achievement tests under one of three test conditions: (1) nonhumorous, (2) low-humor (15% of test items), or (3) moderate humor (30% of test items). These test versions were administered under both low, i.e., short quiz, and high, i.e., examination, outcome-value conditions. Humor frequency did not improve the test performance of highly test-anxious subjects under either outcome-value condition. Together with other previous disconfirmatory findings, the present results suggest that the hypothesized moderating role of item humor in the anxiety-performance relationship may be overstated. Ancillary analyses suggest that individual differences in the use of humor as a coping strategy significantly predict examination scores. PMID- 10477941 TI - Predictive validity of the Young Offender Level of Service Inventory for criminal recidivism of male and female young offenders. AB - 164 male and female young offenders were given the Young Offender Level of Service Inventory. Scores from this 76-item risk assessment were used to produce a matched sample of 82 female and 82 male young offenders. Sex did not influence the 1-yr, criminal recidivism rate, a result consistent with other findings. PMID- 10477942 TI - Parents' and adolescents' perceptions of a strong family. AB - Perceptions of family strengths of 40 parents and 20 adolescents were investigated by means of semistructured interviews and Likert-type ratings on commitment, appreciation, spending time together, communication patterns, religious values, and crisis management. Significant differences were found between the perceptions of parents and adolescents for the family characteristics of commitment, time spent together, and crisis management. Although family strength correlated positively with all six identified characteristics, time spent together and appreciation for each other had the highest correlation with family strength. This result supports the notion that sufficient quality time together may be a prerequisite for well-being in family relationships. PMID- 10477943 TI - Exploring effects of different pictorial stimuli on written expression. AB - This study explored how the type of pictorial stimulus affects the quality of an individual's written expression. Cole, Muenz, Ouchi, Kaufman, and Kaufman in 1997 furnished initial evidence supporting Hooper, et al.'s 1994 theory. A pictorial stimulus different from that used by Cole, et al. was developed from Hooper, et al.'s specifications, i.e., pictorial stimuli should be photographs rather than line drawings, should have a clear protagonist and should present a novel problem situation that can be solved in a stepwise manner and compared to a conventional line drawing stimulus (from PIAT-R Written Expression) in its ability to evoke writing samples. It was hypothesized that the "Hooper" stimulus would yield higher scores than an atheoretical stimulus on items assessing structure and cohesiveness of the story, but not on items that assess writing mechanics. Participants comprised 25 men and women aged 17 to 46 years. Results indicate that Hooper, et al.'s theory is more plausible than a conventional line-drawing stimulus. PMID- 10477944 TI - Pet-assisted psychotherapy. AB - This qualitative study explored the utilization of pets by psychotherapists in their clinical practice with clients who suffer from mental disorders. The sample consisted of 13 master and doctoral level practitioners in social work, marriage and family counseling, psychology, and psychiatry who volunteered to be interviewed regarding their work. The results provide a descriptive overview of the psychotherapists' utilization of pets and their attitude toward this technique. The participants consistently reported that pet-assisted psychotherapy is an effective and efficient technique which can be used with a wide range of clinical problems, age groups, and in diverse practice settings. PMID- 10477945 TI - Use of tests and measures in marital and family research. AB - Measures from the marital and family literature were rank ordered by frequency of appearance in the PsycINFO database from 1974-1997. The Family Environment Scale, Dyadic Adjustment Scale, and the Family Adaptability and Cohesion Evaluation Scales were the instruments found to be most used in research. PMID- 10477946 TI - Sentences and rehearsals increase rated similarity: support for assimilation-in salience theory. AB - The present study concerns how organization produces good retention. Exp. 1 indicated that college students rated two words in sentences as more similar than the same two words alone. In Exp. 2 they rated the two words of old presumably rehearsed pairs as more similar than the two words of new pairs. A sentence is a type of organization, and words that are rehearsed together are frequently subjectively organized. Therefore, the two experiments suggest that organized stimuli also assimilate among themselves (and evidence in the literature concurs). Organized stimuli also produce a highly salient (activated) single memorial group (unit) such as the meaning that a simple sentence comes to have according to evidence in the literature. So, the two experiments support the hypothesis that organized stimuli produce good retention, assimilation among themselves, and a highly salient group. Consequently, organized stimuli may also assimilate to the highly salient group and thereby increase in salience, with this increase enabling the good retention. This assimilation-in-salience theory of how organization produces good retention accords with perceptual evidence suggesting that a target increases in salience through its assimilation to a more salient simultaneously present context. PMID- 10477947 TI - Differentiation of self, need fulfillment, and psychological well-being in married men. AB - The contributions of differentiation of self and need fulfillment within the marital relationship to the experience of psychological well-being were examined in a sample of 95 married men. Differentiation of self, interactional-emotional need fulfillment, and sexual need fulfillment were each associated with higher scores on psychological well-being. Perceived interactional-emotional need fulfillment was identified as the most important predictor of well-being, followed by differentiation of self, and then perceived sexual need fulfillment. Regression analysis indicated that collectively these variables accounted for a significant proportion (27%) of the variance in well-being within the sample. Men who perceived their marital partners to be meeting their interactional, emotional, and sexual needs and who are able to maintain interdependent relationships with their partners were more likely to experience positive mental health. The results point to the importance of attending to issues of self differentiation and perceived need fulfillment within the context of the marital relationship to facilitate psychological health in men. PMID- 10477948 TI - Dysphoric mood and preference for immediate versus delayed monetary reinforcement. AB - It has been proposed that depression is the product of deficits in self management skills: self-monitoring, self-evaluation, and self-reinforcement. While interventions based on this theory have shown promise, some of the basic tenets upon which the theory is based lack empirical support. The present experiment tested one such tenet--the claim that depressed individuals select smaller more immediate reinforcers (an impulsive choice) at the expense of larger more delayed reinforcers (a self-control choice). Currently, empirical support for this notion is sparse and contradictory. This study addressed several methodological problems in earlier studies by creating divergent groups based on Beck Depression Inventory scores, employing a task requiring multiple responses and applying a quantitative model to determine reinforcer value. Analyses indicated no systematic difference between participants in the dysphoric and nondysphoric groups in ability to delay reinforcement. Thus, the current results provide no support for the hypothesis that the 36 dysphoric individuals were unable to delay reinforcement relative to the 21 nondysphoric individuals. Because respondents across the sample as a whole showed a self-control preference, however, the data are consistent with findings in the experimental study of choice responding with adult human subjects. Interpretations in terms of sensitivity and pseudosensitivity to the experimental contingencies are explored. PMID- 10477949 TI - The personal characteristics of effective counselors: what 10 experts think. AB - The counseling professional has called for the use of personal characteristics to be used for admissions to counseling programs as well as in the evaluation of counseling students. 10 expert counselors ranked 22 personality characteristics of potential students for importance and responsiveness to training. The most important include empathy, acceptance, and warmth, while the least important include resourcefulness, sympathy and sociability. PMID- 10477950 TI - Knowledge of laws regulating issues related to HIV and AIDS: development of the HIV/AIDS and the law scale. AB - 42 middle-aged and older adults, ranging in age from 51 to 85 years, completed 10 items dealing with the assessment of knowledge regarding laws regulating issues related to HIV and AIDS. Participants also completed 40 items involving knowledge of risks for HIV infection. The Cronbach coefficient alpha and test-retest reliability coefficient on the HIV/AIDS and the Law Scale were .74 and .83, respectively. Over-all, the grand mean for correct answers was 46.9%, whereas the grand means for incorrect answers and "don't know" responses were 13.6% and 39.5%, suggesting substantial lack of knowledge of laws regulating issues related to HIV and AIDS. Women (50%) and younger participants (51 to 66 years old; 48.2%) showed more of this knowledge (50%) than men (43.0%) and older participants (46.2%). The sample reported a substantial amount of knowledge regarding HIV transmission assessed with factual (92.2% correct) and misconception (87.5% correct) items. The correlation between this knowledge and knowledge of laws regulating issues related to HIV and AIDS was .42 (p < .01). Research with this scale using adolescents and young adults as well as the utility of the scale in areas of clinical, legal, and policy development are discussed. PMID- 10477951 TI - Verbal ability predicts abstinence from drugs and alcohol in a residential treatment population. AB - Measures of cognitive ability, depression, anxiety, antisocial personality, as well as length, type and severity of addiction were obtained from 122 substance abusers enrolled in residential treatment programs. Over a subsequent 6-mo. monitoring period, relapse to substance use was detected in 46 subjects. 17 subjects withdrew from treatment for other reasons and their relapse status was unknown. The remaining 59 maintained abstinence from alcohol or drug use throughout the monitoring period. The only variables to differentiate the groups significantly on outcome were IQ and the Verbal subtest from the Shipley Institute of Living Scale. Stepwise discriminant function analysis indicated that the Verbal component alone correctly identified 64.4% of patients who would successfully remain abstinent. PMID- 10477952 TI - Anti-Semitic prejudice in adolescence: an Italian study on shared beliefs. AB - A study was conducted in Bologna and Venice on a sample of 427 subjects to verify whether anti-Semitic prejudice is observed in Italy, if it has direct, violent expression (blatant prejudice), or if it assumes less evident forms (subtle prejudice). The sample was chosen to represent levels of contact with the Jewish community to check the contact hypothesis (Bologna for the no-contact condition and Venice for the contact-condition) in groups of different ages (adolescents, young people, and adults). We predicted a decrease in blatant prejudice by subjects living in contact with Jews, but no differences in subtle prejudice. Also adolescents, more sensitive to polarised choices, could be less tolerant towards Jews. One-way analysis of variance indicated evidence for anti-Semitic prejudice, whose typical expression was a lack of positive emotions towards Jews. In particular, the whole sample denied negative emotions, such as disregard, annoyance, and irritation, and did not report positive emotions, such as attraction or curiosity. PMID- 10477953 TI - A psychometric analysis of the three gunas. AB - The Vedic Personality Inventory was devised to assess the validity of the Vedic concept of the three gunas or modes of nature as a psychological categorization system. The sample of 619 subjects included persons of varying ages and occupations from a middle-size city in southeastern United States, and also of subscribers to a magazine focusing on Eastern-style spirituality. The original 90 item inventory was shortened to 56 items on the basis of reliability and validity analyses. Cronbach alpha for the three subscales ranged from .93 to .94, and the corrected item-total correlation of every item score with its subscale score was greater than .50. Three measures of convergent validity and four measures of discriminant validity provide evidence for construct validity. The loading of every item on the scale is stronger for the intended subscale than for any other subscale. Although each subscale contains congeneric items, the factors are not independent. The nonorthogonality is consistent with Vedic theory. This inventory requires psychometric development and testing cross-culturally as well as to be experimentally implemented in group research and individual assessment. PMID- 10477954 TI - Effects of negative life experiences on phobia onset. AB - BACKGROUND: Conditioning theories, stress theories and social psychological theories each suggest that negative life experiences should influence phobia onset, though the patterns of effects suggested by each type of theory are different. Few previous studies have estimated the effects of a broad enough range of life experiences on onset of multiple types of phobia to evaluate patterns of effects. METHODS: Retrospective data on life experiences and history of phobia from a representative sample of persons 15-54 years old from the US population (the National Comorbidity Survey) are analyzed using discrete-time event history methods. RESULTS: The effects of 12 negative life events and ten chronic childhood adversities on onset of agoraphobia, specific phobia, and social phobia are presented. Three discrete events have unique effects on agoraphobia onset: life threatening accidents, combat in war (for men), and a fire/flood or other natural disaster. Two chronic experiences during childhood have unique effects on specific phobia onset: violence at the hands of one or more adults, and verbal aggression between parents. Sexual assault by a relative and verbal aggression between parents have unique effects on social phobia onset. The effect of sexual assault by a relative on social phobia is confined to women, and to phobias beginning before age 12. CONCLUSIONS: Unpredictable and uncontrollable events that threaten or result in physical harm influence agoraphobia onset. Potentially predictable but difficult to control childhood experiences (e.g., chronic parental violence) influence specific phobia onset. Blame is a likely mediator of the effect of sexual abuse on social phobia. No data on perceptions of predictability and controllability of life experiences, or of blame, were available for analysis. These conclusions are therefore based on speculations about social psychological processes that have been supported by previous research and theory. PMID- 10477955 TI - The prevalence and associates of depressive disorders in the oldest-old Finns. AB - AIM: To describe the prevalence and associates of major depression and minor depression among the Finnish non-demented population aged 85 years and older (n = 339). METHODS: DSM-III-R criteria were used in diagnosing major depression and dementia. Minor depression was diagnosed by the physician in those who did not fulfil the DSM-III-R criteria for major depression, but had still at least two depressive symptoms. In the first phase, cross-tabulation was used to determine relative risks (RR) and their 95% confidential intervals (95% CI). An additive logistic regression model was then used to find the independent associates of depressive disorders. RESULTS: The prevalence of major depression was 8.1% in men and 4.9% in women, and that of minor depression 18.9% in men and 18.5% in women. In men major depression was associated independently with poor physical health and in women with rare contact with family or friends and poor physical health. Minor depression was associated independently with poor physical health and previous myocardial infarction in men and with poor physical health, a poor ability to walk, and smoking in women. CONCLUSIONS: The prevalence of depressive disorders is quite high among the oldest-old Finns. The factors associated with major and minor depression are largely similar. Although the results suggest that psychosocial stress factors affect the development of both major and minor depression in the oldest-old, no conclusions about causality can be made. PMID- 10477956 TI - Diagnosing mental disorders in primary care: the General Health Questionnaire (GHQ) and the Symptom Check List (SCL-90-R) as screening instruments. AB - BACKGROUND: The treatment of mental disorders in Germany is mainly done by primary care physicians. Several studies have shown that primary care physicians have difficulty in diagnosing these disorders. Recently, several self-report questionnaires have been developed that can be used as screening instruments to identify psychopathology in primary care settings and in the community. The aim of this paper was to investigate the screening properties of the General Health Questionnaire (GHQ-12) and the Symptom Check-List (SCL-90-R) in a primary care setting in Germany. METHOD: A randomly selected sample (n = 408) of adult outpatients from 18 primary care offices in Dusseldorf was screened using the German versions of the GHQ-12 and the SCL-90-R. A structured diagnostic interview (SCID) and an impairment rating (IS) were used as a gold standard to which both questionnaires were compared. Test performance was evaluated by receiver operating characteristic (ROC) analysis. RESULTS: We found no difference in the performance of the general scores of the two questionnaires. Both instruments were able to detect cases. Complex scoring methods offered no advantages over simpler ones for the GHQ-12. ROC analysis confirmed that the SCL-90-R subscales "anxiety" and "depression" showed acceptable concurrent validity for the diagnostic groups anxiety and depression (according to DSM-III-R). CONCLUSIONS: GHQ-12 and SCL-90-R appeared to be useful tools for identifying mental disorders in primary care practice and research. The use of GHQ-12 or SCL-90-R, employed as a first step, supplemented by a second-stage interview, may enhance the detection rate of mental disorder in primary care settings. PMID- 10477957 TI - Markers of inappropriate placement in acute psychiatric inpatient care: a five hospital study. AB - BACKGROUND: Previous researchers in the UK have measured the prevalence of the 'inappropriateness' of acute adult psychiatric inpatient placements by eliciting the judgements of health professionals. We extended the methodology by identifying inpatient sub-groups with stable characteristics that were associated with elevated risk of inappropriate placement, and by calculating the level of that risk. METHODS: A 1-day census was taken in five hospitals. Using a structured instrument, consultant psychiatrists recorded judgements on current placements, recommended alternative placements and stated why alternatives had not been used. Judgements were obtained for 100% of the 261 inpatients. Risk ratios and adjusted odds ratios were calculated to identify sub-groups that were at high risk. RESULTS: Eighty-eight inpatients were inappropriately placed, a point prevalence of 34% (95% CI 28-40%). Consultants recorded that services outside hospital, if available, could have supported most of these inpatients as an alternative to at least part of their hospital stay. Diagnostic rather than socio-demographic or administrative characteristics, in particular primary diagnoses of substance-related disorder, personality disorder, anxiety/stress, and schizophrenia with co-morbid substance misuse, were the strongest markers of elevated risk. CONCLUSIONS: The point prevalence of inappropriate placement within the British mental health-care system is considerable. Diagnostic categories are reasonable pragmatic markers. Further research should aim to identify a strong marker of risk within the numerically large sub-group of schizophrenia without substance misuse. PMID- 10477958 TI - Dealing with extreme environmental degradation: stress and marginalization of Sahel dwellers. AB - BACKGROUND: Psychological aspects of environmental degradation are hardly investigated. In the present study these aspects were examined among Sahel dwellers, who live in environments with different states of degradation. The degradation was assessed in terms of vegetation cover, erosion, and loss of organic matter. METHOD: Subjects came from three cultural groups: Dogon (agriculturalists, n = 225), Mossi (agriculturalists, n = 914), and Fulani (pastoralists, n = 844). Questionnaires addressing marginalization, locus of control, and coping were administered. RESULTS: Environmental degradation was associated with higher levels of stress, marginalization, passive coping (avoidance), a more external locus of control, and lower levels of active coping (problem solving and support seeking). Compared to agriculturalists, pastoralists showed a stronger variation in all psychological variables across all regions, from the least to the most environmentally degraded. Women showed higher scores of stress, (external) locus of control, problem solving, and support seeking than men. The interaction of gender and region was significant for several variables. CONCLUSION: It was concluded that environmental degradation has various psychological correlates: people are likely to display an active approach to environmental degradation as long as the level of degradation is not beyond their control. PMID- 10477959 TI - Parenting style in relation to pathogenic and protective factors of Type A behaviour pattern. AB - BACKGROUND: Studies of type A behaviour pattern suggest that it can be promoted as a whole by certain parental rearing styles. However, the association of the different components of the type A behaviour with specific rearing practices has not been clarified. METHOD: The relationship between parents' rearing style and the different type A behaviour components of their children was analysed in a sample of 312 university students. Parental rearing style was assessed with the EMBU, a Swedish measure originally designed to assess one's recollections concerning one's parents rearing behaviour. Type A pattern was measured by the JAS, a self-administered questionnaire that gives the global type A score and three of its components. RESULTS: Hard Driving was related to Rejection and Favouring Subject in males. Speed-Impatience was related to Rejection and Control in both sexes, and Job Involvement was related to Control and Favouring Subject in females. In a discriminant factor analysis in males, Rejection, Control and Favouring Subject on the part of fathers classified correctly 80% of the subjects identified as having high or low Speed-Impatience and the variables of Rejection and Favouring Subject (also by fathers) classified correctly 69.23% of the subjects identified as high or low Hard Driving. In females, Control and Favouring Subject on the part of mothers and low Rejection by fathers classified correctly 70.37% of the subjects with high or low Job Involvement. CONCLUSION: These results suggest that different rearing characteristics are related to the various components of the type A behaviour pattern. PMID- 10477960 TI - Sex differences in depression after widowhood. Do men suffer more? AB - BACKGROUND: This study focuses on sex differences in depression of the widowed. Previous research showed different results in sex differences and in depression after bereavement. We assessed the effects of widowhood on depressive symptoms for men and women and examined whether environmental strain like social support, finances and housekeeping concerns explain these effects. METHODS: Data were used from a large community-based study of older people in three regions of the Netherlands. Our study sample consists of 2626 widowed and married subjects in the age group of 55-85 years. Depression was measured using the CES-D scale; the various strains were obtained by structured interviews. Multiple linear regression, performed for men and women separately, were used. RESULTS: The results show that widowhood is associated with higher levels of depressive symptoms and that this association is stronger for men than for women. The effect of widowhood is mediated by different types of environmental strain for men and women. However, a strong direct main effect of widowhood on depression remains. The difference in depression rates between men and women is most evident among those widowed for a longer period of time. CONCLUSIONS: It appears that, over time, women adapt to widowhood more successfully than men. From a clinical point of view this is important, as it suggests that men who remain alone after losing their partner are at a higher risk of developing symptoms of chronic depression. PMID- 10477961 TI - [Should utilization of blood in Denmark be changed?]. PMID- 10477963 TI - [Polyneuropathy in critically ill patients]. PMID- 10477962 TI - [Artificial blood]. PMID- 10477964 TI - [Chronic compartment syndrome in the lower leg]. AB - Chronic compartment syndrome (CCS) is characterized by recurrent pain in and functional disability of affected muscles, and is typically encountered in the lower leg. The condition is due to physical activity causing an increased intracompartmental pressure (ICP), which subsequently impedes blood flow and thereby compromises the metabolic demands of the tissues within the compartment. After ceasing of activity the symptoms gradually disappear. The incidence of CCS is uncertain. Diagnosis can be established by measuring the ICP before and after relevant physical activity. Treatment is either restriction in activity or fasciotomy, which in most cases is curative. PMID- 10477965 TI - [Carriers of cystic fibrosis are more susceptible to asthma. The Osterbro study]. AB - We tested the hypothesis that individuals heterozygous for the common cystic fibrosis delta F508 mutation are at risk of obstructive pulmonary disease. We studied a cross-sectional sample from the general population of Copenhagen, aged 20 years and older. We performed spirometry to measure FEV1 and FVC, and genotyped blood samples from 9141 individuals. We identified 250 carriers of the delta F508 mutation (2.7%; 95% CI: 2.5%-3.1%). Nine precent of carriers reported having asthma compared with 6% of non-carriers (chi 2: p = 0.04). Furthermore, among individuals with airway obstruction, the percentage of predicted FEV1 and FVC were significantly lower in participants heterozygous for delta F508 than in non-carriers (49% vs. 58%, p = 0.004 and 70% vs. 82%, p < 0.001, respectively). Cystic fibrosis delta F508 heterozygosity may be over-represented among people with asthma and seems to be associated with decreased pulmonary function in people with airway obstruction who also have asthma. PMID- 10477966 TI - [A quality assessment of notified occupational diseases submitted to the National Occupational Environment Service]. AB - The aim of this study was to assess the quality of a consecutive sample of occupational disease notifications submitted to the National Working Environment Service during 1994. The sample consisted of 860 notifications describing occupational diseases among persons working in companies situated in the county of Vejle. The data information e.g. company name and address, time of employment, harmful exposure and disease, were registered. An overall data quality assessment was performed including evaluation of the etiological connection between described occupational exposure and disease and potential preventive perspectives. The study showed that the notified informations in general were adequate, but doctors need to pay more attention to dose description of the harmful exposure. About 80% of the notifications presented an adequate connection between occupational exposure and disease. Only half of the notifications described preventable and recent (less than five years) harmful exposure. In conclusion, the Danish occupational disease notification system is in general of a high standard, and the National Working Environment Service could make more use of doctors' information provided in these notifications. PMID- 10477967 TI - [Detection of Chlamydia trachomatis infection among young people. The effect of home-sampling and mailing the samples]. AB - Genital Chlamydia trachomatis infection is common and treatable, but often asymptomatic. New diagnostic strategies targeting healthy individuals are therefore needed. The 17 high schools in Aarhus County, Denmark, comprising 8909 students, were cluster-randomized into an intervention group which was offered examination by mailing a urine sample (males and females) and a vaginal flush sample (females) directly to the laboratory, and a control group who were offered examination by a physician. Ninety-three percent (867/928) of the sexually experienced female responders underwent examination in the intervention group, compared with 8% (63/833) in the control group (p < 0.001). The corresponding figures for males were 97%, (430/442) and 2% (4/246), respectively (p < 0.001). Also statistically significantly more infected females and males were found in the intervention group (43 females and 11 males in the intervention group vs. five females and one male in the control group). Home sampling improves the diagnosis of C trachomatis infection among apparently healthy young individuals. PMID- 10477968 TI - [Intraabdominal hemangiopericytoma. Diagnosed during surgery for suspected acute appendicitis]. AB - A 39 year-old febrile man with lower abdominal pain and peritoneal reaction was operated on suspicion of acute appendicitis. During the past eighteen months he had been hospitalized twice due to similar symptoms, both times with spontaneous remission. The operative finding was a large multilobular vascular tumour which on pathological examination was a haemangiopericytoma, with no degree of malignancy. The tumour was removed in toto. PMID- 10477969 TI - [Abscess in the right flank--a late complication of laparoscopic cholecystectomy]. AB - Laparoscopic cholecystectomy (LC) is the treatment of choice in cases of symptomatic gallstones. A 74-year-old female presented with a spontaneously perforated abscess in her right flank and passage through it of gallstones. Four years previously during LC the gallbladder perforated and 10-15 stones were lost intraperitoneally. In case of perforation of the gallbladder during LC it is essential to reduce loss of and remove all gallstones. In case of abdominal symptoms postoperatively, an intra-abdominal abscess must be suspected. PMID- 10477970 TI - [Data collection and analysis]. PMID- 10477971 TI - [Screening for lung cancer]. PMID- 10477972 TI - [Scientific reporting--any future development?]. PMID- 10477973 TI - [One should be careful to comment on something one doesn't know anything about]. PMID- 10477974 TI - Adhesive bonding of fractured anterior teeth: effect of wet technique and rewetting agent. AB - PURPOSE: To compare the fracture strength of intact teeth with that of fractured crowns bonded with adhesive and a resin material on either wet or dry dentin, or dried dentin rewetted with a rewetting agent for various periods of time. MATERIALS AND METHODS: Sheep central incisors were divided into seven groups, each of 10 teeth. Group 1: Intact teeth tested for fracture strength in an Instron testing machine and used as controls. Groups 2-7 were sectioned in a vice equipped with sharp blades, 2.5 mm away from and parallel to the incisal edge. The two fracture surfaces were etched with 37% phosphoric acid for 15 s and rinsed with water for 5-6 s. Group 2: The surfaces were air-dried for 2 s. Group 3: As for Group 2 but the surfaces were rewetted with Aqua-Prep for 2 s and immediately air-blasted to remove excess. Group 4: As for Group 3, but the rewetting agent was applied for 10 s before air-blasting. Group 5: As for Group 4, but the rewetting agent was applied for 20 s. Group 6: As for Group 4, but the rewetting agent was applied for 30 s. Group 7: As for Group 2, but the surfaces were left moist after etching and rinsing by gently shaking the specimens twice. The fracture surfaces of Groups 2-7 were rebonded with All-Bond 2 adhesive system and Aeliteflo composite. After 48 hrs the restored teeth were debonded in the Instron testing machine. Fracture strengths were calculated and compared with the fracture strength of intact teeth (control group). RESULTS: Newman-Keuls' multiple range test revealed that there was a statistically significant difference between the control mean and the means of Groups 2 and 3. All other means were not significantly different. Drying of the dentin surface decreased the bonding strength, and rewetting of the dried dentin for 2 s was not sufficient to rewet the collapsed collagen fibers, as indicated by a decrease of fracture strength. PMID- 10477975 TI - Secondary caries adjacent to amalgam restorations lined with a fluoridated dentin desensitizer. AB - PURPOSE: To determine the in vitro effect of a fluoridated dentin desensitizer solution (DentinBloc) on (1) secondary caries formation, and (2) the interfacial adaptation when used as a cavity liner before amalgam (Dis-persalloy) placement. MATERIALS AND METHODS: 24 macroscopically caries-free molars were selected. Class V cavity preparations on mesial and distal surfaces were placed along the cemento enamel junction, providing an enamel margin and a root surface margin. DentinBloc was applied prior to alloy restoration in the experimental group for 60 s (the excess was removed with cotton pellets), while a copal cavity varnish (Copalite) was used before alloy restoration in the control group. Polarized light evaluation was performed on longitudinal sections (24 caries risk sites for enamel and dentin margins in each group). Alloy-tooth interfaces were evaluated by SEM. RESULTS: Wall lesions were present in 100% of caries-risk sites for the control group, while 58% of enamel risk sites and 50% of root risk sites had wall lesions in the DentinBloc group. Surface lesion depth was reduced 13% at enamel margins and by 18% at root margins in the experimental group when compared to the control group. Relatively small interface gaps between the alloy and tooth surfaces were seen in both groups, but these were less for the root surface margins (2-5 microns) than for the enamel margins (5-15 microns). PMID- 10477976 TI - Efficacy of a triclosan/NaF dentifrice in the control of plaque and gingivitis and concurrent oral microflora monitoring. AB - PURPOSE: To compare the effect of a dentifrice containing 0.3% triclosan and 1100 ppm fluoride and a control dentifrice containing 1100 ppm fluoride on plaque, gingiva and the oral microflora in a long-term study simulating clinical usage. MATERIALS AND METHODS: 159 subjects entered the clinical study and 80 were randomly selected to participate in the microbiological evaluation. 71 subjects completed the detailed evaluation of the oral microflora after 6 months use. Plaque was collected at baseline, 3 months, and 6 months, and examined by darkfield microscopy, Gram stain, immunofluorescence, and selective and non selective media. Changes in antimicrobial susceptibilities were determined for the first 6-month period and for 6 months post-therapy for 68 subjects who completed the entire study. Susceptibilities of whole plaque samples and MIC values for two pre-designated common plaque organisms, A. viscosus and V. parvula were performed. RESULTS: Multivariate ANOVA and non-parametric analyses revealed no statistical differences for any factor tested. No detrimental shifts were found in either; (1) the compositional make up of the normal flora, (2) the periodontopathic or cariogenic flora, or (3) the opportunistic flora in either group of dentifrice users. Both treatments resulted in decreases in Gram positive cocci over time. There was a reduction in spirochetes in the triclosan/fluoride group as compared to the control group. No overgrowth in opportunists, periodontal pathogens, or cariogenic flora was found in either group. No increase in the proportion of the whole plaque flora resistant to triclosan was found nor was an increase in the MIC values of either A. viscosus or V. parvula in either group. Overall, there appeared to be a general decrease in plaque bacteria in both groups over the course of the experiment. PMID- 10477977 TI - A testing system for electric toothbrushes. AB - PURPOSE: To assess in vitro the contributions of mechanical and acoustic microstreaming forces around electric toothbrushes to remove cheese using an experimental model. MATERIALS AND METHODS: Four toothbrushes, Braun Oral-B Plaque Remover (D7), Braun Oral-B Ultra Plaque Remover (D9), Interplak (IP) & Sonicare (SC), were investigated. A model system consisting of a layer of 0.2 mm thick cheese was applied to a microscope slide. The toothbrushes were operated in contact with the slide under loads of 0.3 N dry or with the bristles immersed in 2 mm of water. Turbulence around the moving head was assessed with the bristles placed non-contacting, 1 mm above the slide. An experiment with contact for 2 seconds and then 10 seconds non-contact was also made. The slides were stained and image analyzed. The area of removal was measured together with the "average cleaning/brush contact unit area", (the area of removal was divided by bristle contact area. RESULTS: The removal pattern varied. D9 and SC were efficient when operated dry. When the brushes were operated in water, the D9, SC, and IP were efficient in removing the cheese. With no contact, SC produced disruption of the cheese layer while the other brushes did disturb the surface but did not remove completely the cheese. This disturbance without removal was not recorded by image analysis. When the vibrating brushes were allowed to touch the medium and then moved away, a large amount of removal occurred. Larger amounts of removal took place with the D7, D9 and the SC. The use of brushes with a larger head and bristle contact produced a larger area of removal. However, when the "average cleaning/brush contact unit area" was used, the smaller headed brushes produced a larger amount of removal relative to their size. These differences in the removal characteristics between the four electric toothbrushes in vitro suggest that such a model system may prove useful in testing such devices before they are assessed clinically. PMID- 10477978 TI - Ceramic surface preparations for resin bonding. AB - PURPOSE: To evaluate the effects of hydrofluoric acid (HF) and acidulated phosphate fluoride (APF) primers on differently prepared leucite-containing feldspar ceramic surfaces. MATERIALS AND METHODS: Discs of a leucite-containing feldspar ceramic were prepared following the manufacturer's instructions. Surfaces were either polished, grit blasted or prepared with a diamond bur and then subjected to a range of etching procedures using HF or APF. The surface topography was examined using scanning electron microscopy (SEM). Back-scattered imaging, EDS and X-ray diffraction (XRD) were used for microstructural and compositional analysis. RESULTS: The etching process with HF consisted of a preferential attack of the leucite phase relative to the glass but also removed all features of the previous surface preparation. In contrast, APF etching resulted in the formation of surface deposits, superimposed on the surface roughness created by the prior surface preparations. It was not possible to identify the deposits after APF etching using XRD. PMID- 10477979 TI - Bonding efficiency and interfacial morphology of one-bottle adhesives to contaminated dentin surfaces. AB - PURPOSE: To evaluate the effect of dentin contamination with water, saliva or blood on the bond strength of one-bottle dentin adhesives and interfacial morphology. MATERIALS AND METHODS: The tested materials were Scotchbond 1, One Step, Prime & Bond 2.1 and Syntac SC. The three-step bonding system Scotchbond Multi-Purpose Plus (SMP) was also included as control. Extracted human molars were used. For each tooth, enamel was removed by sectioning with a diamond saw and a smear layer was created by grinding on wet 600 grit SiC paper. Before the adhesive materials were applied and cured, the dentin was dried with air for 3 s, dried with a damp cotton pellet, or contaminated with saliva or blood. Cylinders of the composite resin Z100 were applied onto the adhesive dentin surfaces using Teflon split molds. After 24-hour water storage, the shear bond strength of the specimens was determined using an Instron testing machine at a crosshead speed of 0.5 mm/min. The morphology of the dentin-resin interface was examined using SEM. RESULTS: Wetting of the substrate significantly increased the bond strength produced by One-Step, while the bond strength of Syntac SC was significantly decreased. The bond strength produced by Scotchbond 1, Prime & Bond 2.1 and SMP showed a slight increase or decrease, which was not significant. Saliva contamination decreased the bond strength significantly of only Syntac SC, while blood contamination significantly decreased the bond strengths of all materials. SEM evaluation of the dentin-resin interface showed a relation between resin tag formation and bond strength. PMID- 10477980 TI - Effects of desensitizing agents on human dentin permeability. AB - PURPOSE: To compare the in vitro effects of three desensitizing agents on hydraulic conductance of human dentin: Protect (n = 10), Gluma Desensitizer (n = 10), MS Coat (n = 10) (Pain-Free in the USA). MATERIALS AND METHODS: Dentin discs were prepared from 40 freshly extracted normal human third molars. The pulpal side of the dentin discs was etched with 37% phosphoric acid for 15 s and then rinsed under tap water. The coronal side was sequentially ground and the dentin discs were sonicated for 30 min. The hydraulic conductance was measured filtering 20% serum in phosphate buffered saline under a pressure of 15 cm H2O. The hydraulic conductance of each dentin specimen was measured before using the desensitizing agent and this value was designated as 100%. Thirty dentin discs were treated, the hydraulic conductance was remeasured and expressed as a percentage of the hydraulic conductance of that specimen before treatment. The teeth were stored for 1 month at 37 degrees C in deionized water and the hydraulic conductance of the 40 dentin discs was recorded again. Ten dentin discs were left untreated to serve as a control. RESULTS: No statistical difference was found between the immediate hydraulic conductance of the three groups after treatment. After 1-month storage, the control group showed a statistically higher hydraulic conductance than the three treated groups. There was no statistical difference between the three dentin desensitizing agents evaluated. PMID- 10477981 TI - Ultra-morphological study of the interaction of dental adhesives with carbamide peroxide-bleached enamel. AB - PURPOSE: To evaluate the effects of a carbamide peroxide bleaching agent on interfaces formed by two one-bottle dental adhesives to etched enamel. The null hypotheses tested in this study were that vital bleaching with a commercial 10% carbamide peroxide gel would not (1) increase the concentration of oxygen in the superficial layer of enamel or (2) induce ultra-morphological changes in resin enamel interfaces. MATERIALS AND METHODS: Five extracted human incisors were treated with 10% carbamide peroxide (Opalescence) for 4 h/day for 1 week and were compared with non-bleached teeth for oxygen, calcium, and phosphorus relative concentration using EDS. Mean elemental concentrations were analyzed using a t test (bleached vs. unbleached enamel), one-way ANOVA (for surface location and also for depth) and three-way ANOVA (with bleaching treatment, surface location, and depth as the main factors). For TEM, fifteen extracted human molars were sectioned to obtain two crown halves. After roughening the occlusal surface, one half was bleached with Opalescence while the other half was stored in artificial saliva for 1 week. Enamel was etched for 15 s with a 35% phosphoric acid and bonded with one of three adhesives (Prime & Bond 2.1, Syntac Single-Component, or Scotchbond Multi-Purpose Adhesive-control) and a composite resin (Protect Liner F). Small enamel/resin sticks with a cross-section of 1.0 mm x 1.0 mm were removed and the specimens were processed for TEM observation. RESULTS: Vital bleaching with 10% carbamide peroxide caused no significant changes in relative oxygen concentration of enamel. For calcium and phosphorus, bleaching resulted in significant decreased relative concentrations. Bleaching also resulted in morphological alterations in the most superficial enamel crystallites. Some altered crystallites exhibited electron-lucent cores and reduced thickness of material around the core. PMID- 10477982 TI - Esthetic restoration of primary incisors. AB - A simple and esthetic technique for restoring cariously involved primary maxillary incisors is described. The technique includes mini-pins, a preformed celluloid crown and resin-based composite. PMID- 10477983 TI - 2-year clinical performance of a resin-modified glass ionomer sealant. AB - PURPOSE: To compare the 2-year clinical performance of an experimental resin modified glass ionomer cement sealant (K-512 = Fuji III LC) with that of a light cured resin-based sealant (Delton Opaque) in young adults: MATERIALS AND METHODS: 14 subjects with 47 K-512 and 41 Delton sealants were recalled at 2 years for clinical examination, photographs after enamel etching, impressions and radiographs. RESULTS: K-512 showed 0% complete retention, 62% partial retention, and 38% nil retention. The corresponding percentages for Delton were 32%, 58% and 10%, respectively. There was one instance of fissure caries for K-512 and three for Delton. Sealants deemed to need retreatments because of retention failures were 62% for K-512 and 34% for Delton. The K-512 sealants continued to darken over the study, many becoming slightly darker than the sealed teeth. PMID- 10477984 TI - Retention and caries prevention of Vitremer and Ketac-bond used as occlusal sealants. AB - PURPOSE: To assess the retention and caries incidence of Vitremer (resin-modified glass ionomer) and Ketac-bond (conventional glass ionomer cement), used as occlusal sealants, after 6 and 12 months placement. MATERIALS AND METHODS: The sample consisted of 200 school children, 6-8 years old with sound and unsealed permanent first molars. They were divided into three groups: (a) control group (n = 432 teeth); (b) Vitremer experimental group (n = 200 teeth), and (c) Ketac-Bond experimental group (n = 200 teeth). In the experimental groups, Vitremer was applied on teeth 16 and 46 whereas Ketac-Bond was applied on 26 and 36. RESULTS: The total retention rates for Vitremer after 6 and 12 months were 59% and 36%, respectively. For Ketac-Bond, the total retention rates were 24% and 15%, respectively. No dental caries was recorded during the 12 months for both experimental groups. When effectiveness was measured by sealant retention, there was a significant difference between Vitremer and Ketac-Bond after 6 (P < 0.01) and 12 months (P < 0.05). When effectiveness was measured by caries prevention, no significant difference was found between experimental groups, even when the sealants were partially or totally lost. There was no development of carious lesions after 1-year placement of sealants, but there was a significant difference between experimental and control groups (P < 0.01). PMID- 10477985 TI - Zinc effect on the in vitro formation of calcium phosphates: relevance to clinical inhibition of calculus formation. AB - PURPOSE: To determine the effect of zinc on the in vitro formation of calcium phosphates and its relevance to calculus inhibition. MATERIALS AND METHODS: Different types of calcium phosphate phases (amorphous calcium phosphate, ACP; dicalcium phosphate dihydrate, DCPD; octacalcium phosphate, OCP; and carbonate hydoxyapatite, CHA) were precipitated from solutions containing increasing concentrations of zinc (Zn) ions. The precipitates were characterized using x-ray diffraction, infrared absorption spectroscopy, and scanning electron microscopy. RESULTS: The presence of Zn ions affected the type and amount of calcium phosphate phases formed. Zn, even at concentrations as low as 0.1 mM/L, inhibited the crystal growth of DCPD, OCP and AP; and, at higher concentrations (0.5 mM to 2 mM/L), promoted the formation of amorphous calcium phosphate, ACP, or Zn substituted tricalcium phosphate (beta-TCP) depending on the reaction pH and temperature. PMID- 10477986 TI - Clinical efficacy of two dentin desensitizing agents. AB - PURPOSE: To compare the clinical effect over 3 months of two commercially available desensitizing agents on the level of discomfort of patients with complaints of dentin sensitivity. MATERIALS AND METHODS: An oxalate-containing pre-polymerized resin suspension (Pain-Free), a 0.7% fluoride solution (DentinBloc), and a distilled water placebo were compared in a clinical setting. Ten volunteer patients exhibiting three or more teeth that were sensitive to touch and to a stream of forced air were enrolled in the study (52 teeth total). Each patient's level of sensitivity to tactile stimuli and to a forced air stream was recorded using a visual analog scale (VAS). The tactile stimulus was a metal probe rubbed across the exposed dentin with a constant pressure of 40 grams. A 1 second blast of air from a dental unit air syringe was used to generate the air stream. The desensitizing agents were applied according to manufacturers' guidelines. Sensitivity measurements were recorded at baseline, immediately after treatment, at 1 week, 1 month, and at 3 months. VAS pain scores were analyzed using a repeated measures ANOVA (alpha = 0.05). RESULTS: The pre-polymerized resin suspension (Pain-Free), the fluoride-containing solution (DentinBloc), and the placebo all decreased dentin sensitivity. No significant difference was found among the desensitizing agents at any time period (P > 0.05). PMID- 10477987 TI - Standardized microscopic evaluation of the bonding mechanism of NRC/Prime & Bond NT. AB - PURPOSE: To investigate the in vitro formation of hybrid layer, resin tags and adhesive lateral branches, when using Prime & Bond NT (PBNT) "one-bottle" enamel dentin bonding system on conditioned and unconditioned human dental substrates. MATERIALS AND METHODS: The enamel dentin adhesive was tested on 18 dentin and 18 enamel flat preparations made on facial and occlusal surfaces, respectively, of extracted posterior teeth. Each experimental tooth substrates were randomly divided in six groups of six samples each: Group 1: PBNT on conditioned dentin (CD) with 36% phosphoric acid (PA), Group 2: PBNT on etched enamel (EE) with 36% PA, Group 3: PBNT on CD with NRC non-rinse conditioning (NRC); Group 4: PBNT on EE with NRC, Group 5: PBNT on dentin without any conditioning (UD), Group 6: PBNT on enamel without any conditioning (UE). In the experimental groups, PBNT was applied following the manufacturer's instructions. Over the PBNT, a layer of Dyract AP was placed and light-cured for 20 seconds. All samples were split fractured along their long axis. One half of each sample was deproteinized and decalcified at the interface in order to visualize the hybrid layer and the other was completely dissolved in order to observe the morphology of resin tags with a scanning electron microscope. RESULTS: Groups 1, 2: The tested samples showed a micromechanical bonding mechanism to CD and EE with phosphoric acid. PBNT formed hybrid layer, resin tags and adhesive lateral branches on dentin. In the dentin samples of the first group, characteristic reverse cone-shaped tags with their corresponding adhesive lateral branches were evident. At the enamel site, traditional pattern of etch enamel was always observed with resin tags formation. Groups 3, 4: When the bonding system was applied on dentin conditioned with NRC, a thin hybrid layer was formed, resin tags were also noted but their shape was narrower than that observed in Group 1. At the enamel site, the conditioned surface showed presence of an etched pattern and of resin tags. Groups 5, 6: When PBNT was applied without any previous conditioning, the hybrid layer and resin tags formation were not evident on dentin, and no bonding mechanism was developed with unetched enamel. PMID- 10477988 TI - Debonding of adhesively restored deep Class II MOD restorations after functional loading. AB - PURPOSE: To assess debonding due to functional loading at the cervical dentin margins of deep Class II MOD restorations prepared with a variety of adhesive restoratives in endodontically-treated teeth. MATERIALS AND METHODS: MOD preparations and endodontic treatment were carried out on extracted sound maxillary premolars. The cavities were restored with seven material combinations: Three different resin-based composites (RBC) (Z100, Herculite XRV or Clearfil RP with their respective bonding systems); two bonded amalgam combinations (Valiant with Superbond D or with Panavia 21) and two sandwich combinations [the RBC Tetric with the polyacid-modified RBC (compomer) Compoglass or Z100 with the resin-modified glass ionomer Fuji II LC]. For each group, half of the samples were exposed to mechanical functional loading and the other half served as control. Imperfect bond formation and debonding due to loading was determined by dye penetration. In addition, the dye penetration scores were correlated with the cusp fracture strengths of similarly prepared restorations of a previous study. RESULTS: After functional loading, the RBC in combination with hybridizing dentin bonding agents showed better preservation of marginal integrity than the sandwich restorations, which in turn performed better than the amalgam restorations (P < 0.05). Debonding correlated with reduction in fracture resistance. Under the conditions of this study, debonding of adhesive MOD restorations by functional loading can best be prevented by using RBC in combination with hybridizing dentin bonding systems. PMID- 10477989 TI - Mutans streptococci suppression by chlorhexidine gel in toddlers. AB - PURPOSE: The effect of daily toothbrushing with a chlorhexidine gel on the levels of oral mutants streptococci was evaluated in a group of 37 healthy 1 1/2-year old children. MATERIALS AND METHODS: The microbial samplings were performed with a modified Strip mutans chair-side technique: a sterile cotton pellet on a wooden pin was rolled along the surfaces of the upper primary incisors and then rolled again on a specially prepared plastic strip. The strips were thereafter incubated and cultivated in a selective media. After baseline registrations, the parents were instructed and trained to brush the teeth of their children with a 1% chlorhexidine gel (Corsodyl), twice daily during 14-days. Follow-up samplings were obtained 1 and 3 months after termination of gel treatment. RESULTS: The results revealed a statistically significant (P < 0.01) reduction of mutans streptococci colonization after 1 month compared with baseline. After 3 months, a certain reduction was still evident although not statistically significant from baseline. PMID- 10477990 TI - Bovine dentin as a substitute for human dentin in shear bond strength measurements. AB - PURPOSE: To evaluate the suitability of bovine coronal and root dentin as substitutes for human primary and permanent dentin in shear bond strength tests for dentin adhesives. MATERIALS AND METHODS: 30 bovine permanent central incisors, 30 human primary and 30 human third molars were cut mesiodistally. The pulpal and buccal dentin surfaces of the buccal cuts were milled with a bur to a thickness of 1.0 mm. A dentin adhesive and a hybrid composite were applied exactly according to the instructions given by the manufacturer on each buccal and pulpal surface, except in primary teeth where only the buccal surface was used. Shear bond strength was determined after 24 hours of storage in an aqueous solution. The results were statistically analyzed using the Wilcoxon- or the Mann Whitney-U-test. RESULTS: There were no differences in shear bond strengths between human permanent dentin and bovine coronal dentin (13.3 +/- 6.1 vs. 15.2 +/- 7.6), and between each pulpal and buccal aspect. Significant differences were found between bovine root dentin and human primary dentin (17.4 +/- 8.3 vs. 7.7 +/- 5.0, P < 0.001). Significant differences were also found between bovine root dentin and human permanent dentin (17.4 +/- 8.3 vs. 13.3 +/- 6.1, P < 0.05), and bovine root and coronal dentin (17.4 +/- 8.3 vs. 15.2 +/- 7.6, P < 0.05). Furthermore, significant differences were recorded between human primary and human permanent dentin (7.7 +/- 5.0 vs. 13.3 +/- 6.1, P < 0.001) and bovine coronal dentin (7.7 +/- 5.0 vs. 15.2 +/- 7.6, P < 0.001). PMID- 10477991 TI - Gingiva, teeth and sea salt. AB - This article presents a review of the effects of sea salt on gingival tissues. The beneficial effects of sea salt are described. PMID- 10477992 TI - Caries-preventive effects of sodium and amine fluoride dentifrices. AB - PURPOSE: To determine the comparative effects of sodium (NaF) and amine fluoride dentifrices, as well as the effect of different silica abrasives used in NaF dentifrices, on caries formation in rats. MATERIALS AND METHODS: Harlan Sprague Dawley mixed sex, weanling, littermate rats were fed a cariogenic diet and inoculated with S. sobrinus strain 6715 to induce caries formation. The dentifrices evaluated contained 0.31% NaF with precipitated silica I as the abrasive system; 0.31% NaF with sodium bicarbonate and hydrated silica; placebo; amine fluoride (0.14%) containing silica (Elmex); 0.31% NaF with precipitated silica II (Sensodyne F); and 0.31% NaF with hydrated silica. Dentifrice slurries were administered 7 days per week for 3 weeks at which time the animals were euthanized and the hemijaws removed, coded and stained with a murexide solution. The recovery of S. sobrinus after the 3-week study clearly showed that all of the six groups were infected with S. sobrinus. The Keyes scoring method was then used to assess the caries formation within the six groups. RESULTS: Caries scores showed that all five experimental fluoride dentifrices were equally efficacious statistically in reducing caries formation when compared to the placebo with reductions ranging from 26 to 43%. PMID- 10477993 TI - Fluoride uptake and release by composites and glass ionomers in a high caries challenge situation. AB - PURPOSE: To evaluate the behavior of composite resins and glass ionomer cements with regard to the uptake and release of fluoride, in a high caries challenge situation. MATERIALS AND METHODS: Standard test specimens of glass ionomer cement (Chelon Fil), a resin-modified glass ionomer (Vitremer), two polyacid-modified composite resins (VariGlass and Dyract) and a composite resin (Heliomolar), were submitted for 14 days to demineralization and remineralization cycles in order to simulate a high caries challenge, while from the eighth day onward, a fluoridated dentifrice solution was applied for 5 minutes twice a day and the daily fluoride release of those materials to the mediums was quantified and compared. Fifteen test specimens were prepared for each material, making up a total of 75. RESULTS: All materials studied, except for Heliomolar from day 4 to day 7 in the demineralizing solution, were capable of releasing fluoride in measurable quantities during the whole experiment. The fluoride amounts released by Chelon Fil, Vitremer, VariGlass and Dyract were significantly higher in the demineralizing solution (ANOVA, P < 0.05) than in the remineralizing solution, during almost the entire experiment. The fluoride amounts released by Heliomolar were significantly higher in the remineralizing solution (ANOVA, P < 0.05) than in the demineralizing solution, during almost the entire experiment. All materials studied were capable of uptaking fluoride from the dentifrice solution and of later releasing it to the solution, maintaining the release relatively constant and at a higher level than that seen between days 5 and 7. PMID- 10477994 TI - Shear bond strength of composite resin and amalgam adhesive systems to dentin. AB - PURPOSE: To determine the effects of several multi-purpose dental adhesive systems on the shear bond strength of both composite resin (Herculite XRV) and amalgam (Tytin) to dentin. MATERIALS AND METHODS: 160 extracted human molar teeth were randomly assigned to 16 treatment groups (n = 10/group), embedded in resin, and ground flat on both the occlusal and apical surfaces to expose a 3.0 mm thickness of dentin. In each specimen, a 3.18 mm diameter channel was drilled through the dentin perpendicular to the ground surface. The cylindrical preparations were treated with one of seven adhesive systems or one of two controls (37% H3PO4 or Copalite varnish) according to manufacturers' recommendation, then restored with either composite resin or amalgam. Specimens were thermocycled and subjected to a punch-out shear force using a materials testing machine. RESULTS: All examined multi-purpose adhesive systems provided increased shear bond strength in comparison to the controls. Although the individual performance of each adhesive system varied with specific restorative materials, OptiBond FL demonstrated the highest mean shear bond strength to both composite resin and amalgam. Mean amalgam shear bond strength was significantly greater than mean composite resin strengths. PMID- 10477996 TI - Reliability of outer lesion secondary caries as a predictor of wall lesions. AB - PURPOSE: To assess the reliability of in vitro developed secondary caries outer lesions as an indicator of presence and extent of wall lesions. MATERIALS AND METHODS: Class I occlusal cavities were cut in 132 extracted premolars: 12 teeth were not restored, 120 restored with one of 20 different restoration combinations of silver amalgam, base and varnish. Specimens were aged in 1% NaCl for up to 1 year where after 80 restored and 12 unrestored teeth were subjected to an in vitro bacterial challenge for 36 days, the other 40 specimens were similarly incubated in an acidified broth (pH = 4.0). Sections were prepared for polarized light microscopy and outer and wall lesion areas, outer lesion width, wall lesion length and enamel thickness measured. Data were subjected to Fisher's exact test and Pearson Correlation. RESULTS: There was a significant association (P < 0.05) between outer and wall lesions in all cariously challenged specimen groups. Sensitivity was high 95-96% but specificity was 26% for acid broth, 68% for restored bacterial and 100% for unrestored specimens. Restoration variables impacted widely on specificity. Correlation values were too low to show clinically relevant associations between wall and outer lesion size. Frequency distribution showed that wall lesions extending beyond the dentin-enamel junction were generally associated with potentially remineralizable outer lesions. PMID- 10477995 TI - Effect of surface conditioning on the shear bond strength of compomers to human primary and permanent enamel. AB - PURPOSE: To compare the bond strength of two polyacid-modified composite resins (Compomer) to primary and permanent enamel with and without acid-etching and to test for a possible relationship between the determined shear strength values and area of cohesive failure within the materials on the bonding interface. MATERIALS AND METHODS: Two compomer materials (Dyract and Compoglass) with their respective one-bottle primer/adhesive agents (Prime & Bond 2.0 and Syntac Single Component) were investigated. The parameters tested were shear strength and area of cohesive type of failure after debonding of specimens with respect to the total area of the bonding interface. RESULTS: Mean bond strength to primary enamel ranged from 4.09 MPa (Compoglass) to 5.6 MPa (Dyract), while in permanent enamel the mean values were between 12.3 MPa (Compoglass) and 14.3 MPa (Dyract), when pretreatment was made with bonding agents only. Generally, acid-etching prior to pretreatment improved adhesion significantly (6.0 MPa and 8.1 MPa in primary enamel and 16.0 MPa and 19.4 MPa in permanent enamel for Compoglass and Dyract, respectively). Over one-half of the variation in the values for cohesive failure area was explained by its linear relationship with shear bond strength. PMID- 10477997 TI - Marginal adaptation of compomers in Class I and V cavities in primary molars. AB - PURPOSES: To evaluate in primary molars (1) the adaptation of three compomers in Class I and V cavities at the cavosurface margins and inside the cavity, and (2) the effect of cavity etching on such adaptation. MATERIALS AND METHODS: The three compomers used were Compoglass, Dyract and Hytac. Sixty exfoliated primary molars were divided for the three compomers, which were subdivided into Class I and V (10 each) and subdivided again into etched and non-etched (5 each). Standard Class I and Class V cavities with the gingival margins at cementum were prepared. The cavities were restored with the three compomers used with or without etching the cavity. The restored cavities were finished and polished. The cavosurface margins were inspected under the light microscope. Then the restored teeth were sectioned into two halves. The cut halves were fixed in glutaraldehyde and the cut surfaces smoothed, polished and inspected under the light microscope for adaptation of the restorative material inside the cavities. One halve of each molar was prepared for SEM. RESULTS: The three compomers showed good adaptation at the cavosurface margin except the non-etched Class V cavities restored with Dyract. Compoglass and Dyract were well adapted inside the cavity better than Hytac. Cavity angles were common sites for adhesive pooling for all materials. SEM examination showed good adaptation of the restorative at enamel and cementum interface especially in etched cavities. The dentin-restorative interface showed consistent hybrid layer with resin tags in the etched cavities. However this layer was inconsistent in the non-etched cavities. PMID- 10477998 TI - Sorption and solubility of soft denture liners after accelerated aging. AB - PURPOSE: To determine the sorption and solubility properties of soft liners in relation to the aging process. MATERIALS AND METHODS: Five soft liners (Flexor, Molloplast B, Ufigel L, heat-cured; Simpa, Ufigel P, cold-cured) were used. The sorption and solubility tests were performed as outlined in the American Dental Association (ADA) specification 12 for denture base polymers. Ten specimens of each material were fabricated. One set of five specimens was subjected to accelerated aging process in the Weather-Ometer instrument. The test materials were dried in a desiccator and then immersed in distilled water at 37 +/- 1 degrees C for 15, and 30 days. The sorption behavior of the test specimens was calculated. The solubility behavior of the test materials was determined only after 30 days. RESULTS: Aged materials immersed in water for 15 days showed different sorption properties related to the material. After water immersion for 30 days, the materials showed increased sorption properties. Negative increase in solubility was determined after aging for all liners except Flexor. PMID- 10477999 TI - Wear of composite resin inlays and antagonistic enamel. AB - PURPOSE: To evaluate the wear of composite resin inlays and antagonistic enamel wear when they opposed each other. MATERIALS AND METHODS: Cylindrical-shaped composite disk specimens were fabricated with three composite resin inlay materials and a heat-treated direct composite resin (control). The lingual cusps of extracted human maxillary premolars were placed perpendicular to flatten composite resin inlay specimens. Two body localized wear test was carried out using six specimens per material. The surface area of worn enamel and composite specimens was measured by a computerized analysis system. RESULTS: There were statistical differences between inlay materials and the control with respect to enamel wear, although no differences existed between the inlay materials tested. Worn enamel analysis by SEM indicated that large size quartz filler particles created the coarsest surface. The results suggest that the composite resin inlay materials tested are better than the quartz-containing composite resin in terms of antagonistic enamel wear. PMID- 10478000 TI - Fabrication of resin-bonded three-unit prostheses. AB - This article describes the fabrication of two fiber-reinforced poly-glass ceramic three-unit fixed prostheses. A second-generation laboratory-processed polymer glass ceramic material (Sculpture) that utilizes a resin-impregnated translucent fiber framework (Fiberkor) was used to fabricate two fixed resin-bonded prostheses. The abutment teeth of one appliance required the use of a full coverage preparation and an inlay design. The other prosthesis required an inlay preparation and onlay configuration. Two three-unit provisional prostheses were fabricated and inserted using eugenol-free temporary cement. With the fabrication of the fixed appliances, they were bonded to the abutment teeth with resin cement. The insertion of the two fixed appliances using the polymer-glass ceramic material provided the esthetic advantage of translucency, while avoiding the appearance of metal. PMID- 10478001 TI - Microbiology of carious lesions. AB - Dental caries is a process in which the enamel, and later the dentine, is demineralized by acids produced by bacterial fermentation of carbohydrates. The main indicator bacteria in this process are mutans streptococci and lactobacilli. This article discusses how monitoring of patients' salivary levels of these organisms can help in prevention and treatment of caries in different patient populations. PMID- 10478002 TI - Recognizing and caring for the medically compromised child: 1. Disorders of the cardiovascular and respiratory systems. AB - This paper describes important disorders of the cardiovascular and respiratory systems and the management of dental treatment for patients with these diseases. PMID- 10478003 TI - Basic computing for dental practitioners: 5. Practice management systems. AB - This article examines the specialist software that is commercially available to assist the running of a dental practice. Generically termed 'practice management systems' (PMSs), these programs are designed to help perform many of the clerical, administrative and accounting tasks traditionally carried out manually, in a shorter time and with greater accuracy. PMSs have benefited considerably from advances in computer technology, particularly the increased speed of processors and graphic displays, to the point that they have become a viable alternative to an appointments diary and clinical records. PMID- 10478004 TI - Dilacerated incisors and congenitally displaced incisors: three case reports. AB - Three cases of dilacerated incisors and congenitally displaced incisors are reported. Dilaceration of central incisors occurs following trauma to the deciduous dentition. Generally these teeth are so severely malformed that they have to be extracted. Congenital displacement of the central incisors is an idiopathic condition with a generally favourable prognosis for orthodontic alignment. PMID- 10478005 TI - Assessment of the radiographic image: recognition of normal features. AB - The importance of observation, and perception, based upon an appropriate level of knowledge, is introduced in this article as the basis for examining dental radiographs. Normal anatomical features are displayed in radiographs as relative radiolucencies or radiopacities, their precise appearance varying with the technique used and exposure factors. This article addresses some of the points related to initial examination of radiographs, and the influence on the image of a variety of anatomical features. The author presents a logical approach to examining radiographs and illustrates the key anatomical features in the region of the jaws. PMID- 10478007 TI - The differential diagnosis of sensitive teeth. AB - A major presenting complaint in dental practice is of sensitive teeth. In order to manage the problem well, a correct diagnosis is essential. Whilst it is true that 'common things occur commonly', one should consider all possible causes of pain in taking a history and examining the patient. This paper summarizes how to identify the potential causes of such a complaint. PMID- 10478006 TI - Emergency drugs in dental practice. AB - An increase in the life expectancy of medically compromised patients has resulted in greater numbers of these individuals within our ageing population. As dental surgeons are increasingly expected to provide care for such patients, the likelihood of a medical emergency occurring within the dental surgery may increase. This article reviews the more common emergency drugs that a dentist may have to administer in the event of a medical emergency. The indications, mode of action, method of administration and dose of each agent are discussed. PMID- 10478008 TI - Recognizing and caring for the medically compromised child: 3. Disorders affecting the gastro-intestinal tract and human immunodeficiency virus infection. AB - Oral manifestations of gastro-intestinal disease are commonly under-diagnosed. This paper is the third in the series entitled 'Recognizing and Caring for the Medically Compromised Child' and seeks to facilitate the process of diagnosis and management of disorders affecting the gastro-intestinal tract in terms of both oral and gut signs and symptoms. This paper concludes with a section devoted to the recognition of HIV disease in paediatric patients, both orally and generally, and its implications for management within the dental surgery. PMID- 10478009 TI - The relationship between obstructive sleep apnoea and dentistry: 2. Management. AB - Obstructive sleep apnoea (OSA) is a potentially life-threatening breathing disorder characterized by repeated collapse of the upper airway during sleep, with cessation for breathing. Patients with OSA are at risk of severe cardiovascular and respiratory complications secondary to recurrent hypoxia and hypercapnia in addition to an overall reduced quality of life for sufferers and their families. An overview of the aetiology including pathophysiology, clinical signs and symptoms and diagnosis was presented in the first article in this series. In this article the management of OSA is presented, with particular emphasis on the role of dentistry in contemporary management. PMID- 10478010 TI - Orofacial disease: update for the dental clinical team: 2. Ulcers, erosions and other causes of sore mouth. Part I. AB - Mouth ulcers are one of the most common oral complaints. The term ulcer is used usually where there is damage to both epithelium and lamina propria, and a crater, sometimes made more obvious clinically by swelling caused by oedema or proliferation in the surrounding tissue. The term erosion is usually used where the damage is somewhat more superficial. Most ulcers/erosions are due to local causes such as trauma or burns. Some are caused by aphthae or malignant neoplasms, and a few have aetiology in obvious systemic disease. Ulcers of local cause, drugs, aphthous ulcers, Behcet's syndrome and malignant ulcers are discussed in this article. The next two articles discuss the ulcers due to systemic disease including disorders of the blood, gastrointestinal disorders, skin diseases, connective tissue disease and infective diseases. PMID- 10478011 TI - A practical guide to antibiotic prophylaxis in restorative dentistry. AB - The purpose of this paper is to review the principles and practicalities of antibiotic prophylaxis in medically compromised patients who require restorative dentistry. Guidelines for the management of patients who are susceptible to infection are given. The importance of oral health and the rational use of both local and systemic antimicrobial prophylaxis is described. PMID- 10478012 TI - First impressions: just what do my patients think of me? AB - The success of a dental practice, in terms of both finance and job satisfaction, depends on effective communication with our patients. This paper highlights the key aspects of communication theory which we should include in our day-to-day practising lives. PMID- 10478013 TI - Recognizing and caring for the medically compromised child: 4. Children with other chronic medical conditions. AB - This is the fourth and final part of a series on recognizing and caring for medically compromised children. In this article, an outline of appropriate dental management for children with other more commonly encountered chronic medical conditions is given, together with a description of the disorders and their significance in dentistry. This group includes children with physically handicapping conditions and children with learning difficulties, as well as those who are medically compromised. PMID- 10478014 TI - Seating of composite inlays with ultrasonic vibration. AB - The use of vibration enables the clinician to use high-viscosity luting agents when seating composite inlays and allows quick and efficient placement of the inlay. The film of composite luting agent is achieved in a much shorter time and with lower loads by ultrasonic vibration than using manual methods. In order to achieve a thin lute, the vibrations of the ultrasonic scaler must be directed perpendicular to the surface of the inlay. PMID- 10478015 TI - Orofacial disease: update for the dental clinical team: 2. Ulcers, erosions and other causes of sore mouth. Part II. AB - This third article in the series discusses mouth ulceration in relation to blood dyscrasias, gastrointestinal disorders, skin and connective tissue disease. PMID- 10478016 TI - Assessment of the radiographic image: recognition of abnormal features. AB - Radiographs, due to their ability to pass through materials differentially, are used in dental practice to obtain information that is not available by other means. Recognition of the presence of a variation from normal is the first step to accurate interpretation of abnormalities. This article presents an outline of a method for examining radiographs in order to recognize that an abnormality is present, and to learn from the visual information some of the characteristics of the abnormality. A number of clinical examples are used to illustrate the conclusions that we can draw from the radiographic changes. PMID- 10478017 TI - The influence of alveolar bone grafting on the orthodontic and prosthodontic treatment of patients with cleft lip and palate. AB - The introduction of mixed dentition bone grafting of alveolar clefts means that the alveolar process can be fully restored, permitting adjacent teeth to migrate or be orthodontically moved into the grafted bone. Thus a complete dental arch can be obtained without prosthodontics in the great majority of patients. In addition, oral-nasal fistulae are closed, mucosal recesses eliminated (facilitating oral hygiene) and the long-term periodontal health of the teeth adjacent to the former cleft is improved. Alveolar bone grafting with subsequent orthodontic treatment, together with advances in dental materials, have contributed substantially to the care of patients with alveolar clefts, reducing the need for prosthodontic procedures and allowing completion of the dental treatment at an earlier age. PMID- 10478018 TI - Recent advances in ceramic materials and systems for dental restorations. AB - The unsurpassed aesthetic and biocompatible qualities of ceramic materials still provide the stimulus to seek to overcome their limitations. Much of the materials research since the mid 1960s has been directed towards producing stronger, reinforced restorations with improved marginal accuracy and, although a new wave of ceramic products appeared in the 1980s, most of the materials currently available are developments derived from ideas established more than 20 years ago. This paper outlines some interesting aspects of recent developments and considers the present state of the art with respect to both materials and techniques. PMID- 10478019 TI - Orofacial disease: update for the dental clinical team: 2. Ulcers, erosions and other causes of sore mouth. Part III. AB - This article discusses the various infective causes of mouth ulcers. PMID- 10478020 TI - Career pathways in dentistry. AB - During the next decade we are likely to see the greatest changes in the way in which dentistry is delivered than at any point since the introduction of the National Health Service in 1948. The move in the primary-care sector from a mainly NHS service to a mixed economy, the emergence of more practices operated by bodies corporate and the possibility of high-street specialists will all impact on every aspect of professional life. For those trying to plan a career or looking to change pathways it can be difficult to identify which pathway to follow. The purpose of this article is to explore the possible routes for career development. PMID- 10478021 TI - Handling of mechanical stresses in composite restorations. AB - The origin of stress in adhesive resin composite restorations is attributed to restrained shrinkage during polymerization and is dependent on the configuration of the restoration. Moreover, non-homogeneous deformations during functional loading can damage the interface as well as the coherence of the material. Damage from these stresses can be reduced by application of an elastic lining at the adhesive interfaces and by slowing the initial conversion by two-step light initiation of the resin. The various factors that mediate flow and compliance are discussed. PMID- 10478022 TI - Microscopic features of clinically successful dentine bonding. AB - Clinical success with resin-based bonding systems depends upon meticulous attention to detail at each stage of the bonding process. Cavity margins placed in enamel wherever possible and etched for at least 15 to 30 seconds will ensure a reliable peripheral seal. Dentine conditioning should be brief--a maximum of 15 seconds for surface demineralization only. Diffusion of primers into the full extent of dentinal demineralization is enhanced by application of multiple coats. Any solvent or moisture remaining on a primed dentine surface will prevent adaptation of bonding resin, resulting in non-attachment. Adhesive bonding resin provides a valuable elastic intermediate layer for absorption of stresses of polymerization shrinkage--but it should be of uniform thickness and not air thinned. Incremental insertion of resin composite will reduce polymerization stresses and ensure a fully cured restoration. PMID- 10478023 TI - Basic computing for dental practitioners: 4. Computer applications--spreadsheets and databases. AB - This article, the fourth in a series, discusses the uses of spreadsheets and databases in the dental practice. PMID- 10478025 TI - Burning mouth syndrome: approach to successful management. AB - Burning mouth syndrome is a fairly common condition. Contrary to popular belief, its management is successful in about 70% of cases--provided a structured protocol based on scientific evidence is adopted. Specialist advice must be sought in some cases, but many patients can be successfully treated in the primary care environment. PMID- 10478024 TI - Are the features of ankyloglossia limited to the lingual fraenum? AB - A 16-year-old male patient presented with ankyloglossia accompanied by a complaint of forceful spurting of saliva from the mouth and a unilateral swelling in the submandibular region. Following surgical relief of the ankyloglossia, the spurting of saliva had virtually ceased and the submandibular swelling disappeared without recurrence. PMID- 10478026 TI - The use of myofunctional appliances in the UK: a survey of British orthodontists. AB - A questionnaire was sent out to members of the British Orthodontic Society in an attempt to establish how myofunctional appliances are currently used in the UK and to attempt to explain the differences between provider groups. This article reports the findings of this survey and that, of all the appliances currently available, British orthodontists prefer to use the Twin Block appliance. Furthermore, most of the sample questioned felt that the laboratory cost of many myofunctional appliances meant their provision under the General Dental Service was not practical. PMID- 10478027 TI - Dental education--25 years on: a personal view. AB - Change in dental practice has been complemented by perhaps even more profound change in dental education at all levels. Not only has the structure been transformed but the underlying philosophy has shifted to an emphasis on lifelong learning as the only strategy for professional survival in the world of exponential knowledge growth and shifting values. PMID- 10478029 TI - Prevalence of dental caries: retrospect and prospect. AB - In spite of the general decline in dental caries, as illustrated in national surveys of children's health and by BASCD surveys, considerable unmet need remains. If good oral health is to be ensured for all sections of the UK community, the issues discussed in this article must be addressed. PMID- 10478028 TI - Fluoride: how to maximize the benefits and minimize the risks. AB - This article reviews current thinking on the rational use of fluoride for the safe and effective prevention of dental caries, so that benefits may be maximized and risks minimized. PMID- 10478030 TI - The relationship between obstructive sleep apnoea and dentistry: 1. Aetiology and diagnosis. AB - Obstructive sleep apnoea is a potentially life-threatening breathing disorder characterized by repeated collapse of the upper airway during sleep, with cessation of breathing. Patients are at risk of severe cardiovascular and respiratory complications secondary to recurrent hypoxia and hypercapnia and the overall quality of life for sufferers and their families may be reduced. This article gives an overview of the aetiology (including pathophysiology), clinical signs and symptoms, diagnosis and treatment. The role dentistry can play in contemporary management of the disorder will be discussed in a second article. PMID- 10478031 TI - Assessment of caries risk. AB - This paper examines the importance of caries risk assessment, identifies relevant risk factors, including clinical evidence, which has been considered to be the best predictor of caries risk. PMID- 10478032 TI - A review of burnout in dentists. AB - Burnout, a distinct psychological construct that has a number of features which separate it from occupational stress, is strongly associated with the emotional effects of providing patient- or client-centred services. Depression may be a consequence of prolonged experience of burnout. Recent findings suggest that burnout has features of maladaptive coping in the short term but is, paradoxically, protective in the longer term. Dentists are prone to burnout due to the nature of their work but may be able to prevent it if they can recognize the burnout process and take regular holiday breaks. Burnout is assumed to have an adverse influence on patient care, although no dental studies, as yet, have tackled the issue. PMID- 10478033 TI - The road to specialization. AB - This paper describes the history and current status of specialization within dental practice in the UK. PMID- 10478034 TI - Recognizing and caring for the medically compromised child: 2. Haematological disorders. AB - Haematological disease in his/her patients is of particular relevance to the dental surgeon; the diagnostic signs and symptoms may be manifest in the mouth, and patients with haematological disorders must be managed carefully. This paper, the second in a series on recognizing and caring for medically compromised children, provides a simple classification of the haematological diseases affecting children that a dental practitioner is likely to encounter. In it, the diagnostic features, including oral manifestations, of these disorders are described and current 'best practice' in the management of both general and oral signs and symptoms is discussed. The implications of the general management of haematological disease to dentistry are also discussed. PMID- 10478035 TI - The future of dental materials. AB - This article reviews some of the rapid developments in dental materials that have occurred during the last 25 years, especially adhesive, biocompatible and aesthetic materials. Some innovative research developments, which have exciting possibilities for future dental treatment, are outlined. PMID- 10478037 TI - Orofacial disease: update for the dental clinical team: 1. Diagnosis and management of orofacial disease. AB - The increasingly ageing population, advances in medical and surgical sciences, and the advent of new diseases such as infection with the human immunodeficiency virus (HIV) have highlighted the importance of orofacial diseases. This article begins a new series on current thinking in the areas of orofacial medicine most relevant to the dental care team. PMID- 10478036 TI - Basic computing for dental practitioners: 6. Multimedia and communication. AB - This final article in the series on computing for dental practitioners examines multimedia and the Internet. Multimedia is one area where the greatest advances have been made in recent years; almost all PCs marketed today have multimedia capabilities and more and more software is being written with multimedia attributes. PMID- 10478038 TI - The incidence of periodontal disease and its impact on future dental practice. AB - This paper reviews the prevalence and possible changes in prevalence of periodontal disease from epidemiological investigations carried out within the previous two decades. Although many older studies used unreliable indices, there is some evidence that clinically significant periodontitis is now restricted to between 15 and 20% of the population with certain groups, such as smokers and diabetics, at particular risk. The outlook for general dental practice in the future is also considered. As elderly people will form a greater proportion of the future population, and are more likely to remain dentate, more periodontal treatment may be required. To identify individuals and communities at risk, high quality periodontal epidemiological information is of more importance than ever before. PMID- 10478039 TI - The personality of birth. PMID- 10478040 TI - Home-like hospital birth. Interview by Marion Toepke McLean. PMID- 10478041 TI - Not among strangers. PMID- 10478043 TI - Homebirth: what are the issues? PMID- 10478042 TI - Why homebirth? PMID- 10478044 TI - Home--our birth right. PMID- 10478045 TI - Healing through homebirth. PMID- 10478046 TI - Out of the womb, into the fire. The myth of the safety of hospital childbirth. PMID- 10478047 TI - Ultrasound: more harm than good? PMID- 10478048 TI - Drugs in labour. What effects do they have twenty years hence? PMID- 10478049 TI - Pelvises I have known and loved. PMID- 10478050 TI - Elaine's birth. PMID- 10478051 TI - A declaration of the rights of childbearing women. PMID- 10478052 TI - The missing squatting bar. Childbirth education in the 90s. PMID- 10478053 TI - 7 recommendations for sister midwives. PMID- 10478054 TI - Mourning the professionalization of midwifery. PMID- 10478055 TI - Four birth stories. PMID- 10478056 TI - A birth in Ecuador. PMID- 10478057 TI - All was well: homebirth in Australia. PMID- 10478058 TI - Clara's birth. AB - Advocacy for homebirth is based on the strong assumption that birthing is a physiological process and does not require medical interventions unless things turn "wrong." Let us assume that something might always go wrong, for instance during Clara's birth when the placenta was still retained after three hours. What needs to be done? The moment the midwife entered the house she was endowed with a responsibility for any problem caused by her failure to give proper guidance. With this weight on her shoulder, and according to her training and experience, there was no other way for her than to suggest an intervention regarding the placenta. The two midwives, B, and C., might not agree on risk estimations, the nature of the intervention, whether it should be performed at home or in a hospital. The estimation of abnormalities, evaluation of risks and the procedures with which to handle them are the main practical difference between classic obstetrics and non-interventionist midwifery--by analogy, between allopathy and naturopathy. The rest (positive thinking) is basically literature. A delivery will not remain normal just because we decide it "must" be physiological. Dr. Barua, a professor of obstetrics in Pondicherry, pointed out that normal deliveries are rare--fewer than 10 percent in South India. What we have instead is either pathological or "natural" deliveries in which regenerative processes take care of abnormal situations. Unless she has developed sensitive hands, a birth assistant or midwife must rely on monitoring procedures to evaluate deviations from the normal process. Even with the greatest care, these procedures are intrusive in that they disconnect the parturient from her own sensations. While successful unattended homebirth stories emphasise the extraordinary power and sensitivity of a birthing woman, the whole dream seems to collapse in abnormal or pathological cases. It would have collapsed for Sonia as well, had she not discarded negative suggestions. Dependency on medical experts is an alienation by which the patient is forced to tolerate what is not tolerable to her at a precise moment. The trauma of medicalized birth is not surprisingly associated with rape. It remains that the leitmotif in homebirth literature is more "empowerment," self assertion, militancy for the freedom of choice and ownership of her own body, than "self reliance" which involves a long-term sensitisation. Even outstanding essays a advocating non-medicalized birth may contain a sentence with implicit fear talk defusing the argument. For instance: "And if need be, [the midwife] can transport you to the hospital if you truly need the help that only modern obstetrics can provide...."] The sentence may be countereffective to a reader who is insecure about homebirth. The problem lies in the proximity of "truly" and "only." It was indeed the writer's intention to emphasise "truly" with an interrogative connotation: "Are you sure you need it?" However, the effect is overruled by the assertive strength of "only." The reader's unconscious reading may be that, truly, she needs the help that only modern obstetrics can provide. This is an example of negative suggestions conveyed by confidence building talk about health and pregnancy. Another example was Denis and Sonia's obsession with the placenta retention problem that had been causally exposed by midwife B. Why should her empowerment end suddenly when there is a life threatening complication during the delivery? If the pregnant woman claims a fundamental right to choose her way of birthing in uncomplicated cases, she needs even more self determination when she is in real danger. This paradox is striking evidence that the common sense attitude toward danger is affected by the "allopathic mind." It sets up an insurmountable barrier between the physiological and the pathological, whereby symptoms are failures of the normal regenerative processes. Hence, testimonies of women feeling that their bodies "b PMID- 10478059 TI - Bringing the old ways back. Interview by Clarebeth Loprinzi-Kassel. PMID- 10478060 TI - Informatics in the NHS: paving the way for advanced practice. PMID- 10478061 TI - Critical appraisal. 4. Questioning evidence: its validity and importance. PMID- 10478062 TI - Education for practice. On course for research awareness. PMID- 10478063 TI - Clinical governance. 2. Organisational effectiveness--2. PMID- 10478064 TI - Healthcare open learning. Empowerment through practice-based learning. 2. The balance of power. PMID- 10478065 TI - Learning organizations ... and other benefits of the Senge culture. PMID- 10478066 TI - Concerted action to meet the nation's nursing needs: the first NLN convention. PMID- 10478067 TI - Integrating advanced practice nurses in home care. Recommendations for a teaching home care program. AB - MUCH, if not all, of the health service needs of the frail elderly can, and will, be provided in the community and at home. To be cared for at home is the overwhelming preference of the growing number of older people with chronic illness and those who are physically dependent-persons "aging in place" in their own domiciles, in naturally occurring retirement communities, and in public or privately funded adult housing residences and assisted living facilities. Medicare and Medicaid capitated financing is increasingly directed at programmatic and reimbursement initiatives that promote "independent living." PMID- 10478068 TI - A collaborative effort for sex education in rural school settings. AB - Many students report that sex and sexuality education is, at best, inappropriate and, at worst, nonexistent. Alarming statistics for teen pregnancy and a high incidence of sexually transmitted diseases support the need for education and intervention targeted at a young audience. The traditional approach, to incorporate sex education into health education classes taught by schoolteachers, can be difficult to accomplish in rural school settings where adequate financial resources and community involvement are lacking. Through a partnership funded by the W. K. Kellogg Foundation, a collaborative approach was used in two rural Southeastern communities to deliver a program of sex education to students in fourth through sixth grades. The program united the efforts of community leaders, the school administration, parents, and students and faculty of a university school of nursing. PMID- 10478070 TI - Community-based nursing education. Research study from the NLN Vision for Nursing Education--Hawai'i/phase II. AB - Four sites--Hawai'i, Minnesota, Texas, and Massachusetts--were selected by the NLN to participate in the National League for Nursing Vision for Nursing Education: Phase II Project, a one-year initiative focused on community-based nursing education. This initiative, for the year 1995-1996, was funded by the W. K. Kellogg Foundation and coordinated by a national project director. Each site received $100,000 to be used to develop curricular models for community-based nursing. Other components of the initiative as it took place in Hawai'i--an exploration of regulatory issues regarding community-based nursing education and a description of the process of change--are reported elsewhere. PMID- 10478069 TI - A learning-service community partnership model for pediatric student experiences. AB - Nurse educators are attempting to revise the nursing curriculum to keep pace with the changing focus of health care and prepare graduates for practice in a community-based, community-focused health care system (1,2). An important aspect of the change in emphasis in nursing education is the need to address the primary and chronic health care needs of individuals, families, and aggregate populations. PMID- 10478071 TI - Focus groups. A unique approach to curriculum development. AB - The purpose of this article is to describe the use and effectiveness of focus group methodology for curriculum development. This unique approach grew from the investigators' belief in the application of the principles of primary health care in designing a curriculum for community-based nursing education. The specific content and outcomes of the focus groups in relation to three curricular themes are reported in this journal in the article "Community-Based Nursing Education: Research Study from the NLN Vision for Nursing Education--Hawai'i, Phase II." The current article concentrates on the process and methods used. PMID- 10478072 TI - How does NLNAC respond to increasing reliance on part-time faculty? PMID- 10478073 TI - The problems with recruitment and retention are not going to go away. PMID- 10478074 TI - It's a bug's life. PMID- 10478075 TI - Safely employed? PMID- 10478076 TI - Restraint or protection? The use of bedside rails. PMID- 10478077 TI - Palliative care for the future. PMID- 10478078 TI - Medical device management. PMID- 10478079 TI - Leading nurses into the future. PMID- 10478081 TI - Trust amalgamation. A partnership challenge. AB - Structure and role clarity should be an early priority A rigorous and fair appointments process is required Short term performance will be affected Communicate all the time The 'must be dones' still have to be done The trust has not reached its first birthday PMID- 10478080 TI - Putting children first. PMID- 10478082 TI - [Need for care--an uncalculable quantity]. PMID- 10478083 TI - [Effects of competition in ambulatory care]. PMID- 10478084 TI - [Dialog on return as a management instrument]. PMID- 10478085 TI - [Publicity work, I. What should be emphasized?]. PMID- 10478086 TI - [Nursing case management in the USA]. PMID- 10478087 TI - [Development of the profession, differentiation of the profession]. PMID- 10478088 TI - [Education and counselling as a nursing mandate]. PMID- 10478089 TI - [The right of the employer to make rules]. PMID- 10478090 TI - [How do we assure and create employment in nursing?]. PMID- 10478091 TI - [A dangerous gap]. PMID- 10478092 TI - [The experts are divided on payable quality]. PMID- 10478093 TI - [Documentation in the hospital: between annoying duty and a reasonable obligation]. PMID- 10478094 TI - [Care of the elderly has to be scientifically supported]. PMID- 10478095 TI - [Urinary diversion by ileal conduit. Qualified stoma care helps to avoid complications]. PMID- 10478096 TI - [Equipment-supported respiratory therapy: patients are trained in weaning before surgery]. PMID- 10478097 TI - [Applying a gastric catheter. Its position has to be checked]. PMID- 10478098 TI - [Patient imports from the Third World: African children are supposed to finance German pediatric facilities]. PMID- 10478099 TI - [Developmental psychology: children need adequate care]. PMID- 10478100 TI - [Development by determination of goals: the direction is determined, not the concrete path]. PMID- 10478101 TI - [Understanding care within the framework of flat rates per case: standards are not just an accounting basis]. PMID- 10478102 TI - [Stopping life-support measures: the patient's intentions have to be ascertained without a doubt]. PMID- 10478103 TI - [Asking nursing students about their choice of profession: are we born to be nurses?]. PMID- 10478104 TI - [Nursing in the 16. century: Regina Protmann--a pioneer in community health care]. PMID- 10478105 TI - ["The English Patient": professional care in times of war]. PMID- 10478106 TI - [Transcultural care. The end of wandering. Ambulatory care of aged working migrants in Germany]. PMID- 10478107 TI - [Making nursing visible]. PMID- 10478108 TI - [More money, fewer positions--employers are threatening with more savings]. PMID- 10478109 TI - [Hospice meeting "Death Wish and Impatience at the End of Life": death makes sense for the sake of life]. PMID- 10478110 TI - ["Dienste fur Menschen" is reacting to negative press: a dean becomes the wailing wall for home residents]. PMID- 10478111 TI - [The Hans Bockler Foundation presents study on nursing insurance: need for care remains a risk factor for the poor]. PMID- 10478112 TI - [Looks, personal feeling and presence in nursing: there is no second chance for a first impression]. PMID- 10478113 TI - [Focus on decubitus ulcers: industry is betting on hydroactive wound dressings]. PMID- 10478114 TI - [Comparing baby diapers: less chaffing with superabsorbers?]. PMID- 10478115 TI - [Nursing in pediatric anesthesia: a child is not a small adult]. PMID- 10478116 TI - [Basal stimulation in intensive care: offering elemental stimulation to the patient]. PMID- 10478117 TI - [Changing a colostomy pouch: the type of ostomy determines the procedure]. PMID- 10478118 TI - [A first model try-out in Germany: common basic training for all nursing professions]. PMID- 10478119 TI - [Conflicts between nurses and physicians: lack of time impedes the flow of information]. PMID- 10478120 TI - [Dialog after the return from absence from the department: it is not just about the reduction of absences]. PMID- 10478122 TI - [Nursing documentation: a legal must or a beneficial duty?]. PMID- 10478121 TI - [Communication with demented patients: patient-centered dialog requires special abilities]. PMID- 10478123 TI - [Health care as an educational subject. Integration of health care aspects into nursing education at the Nursing School of the Giessen Justus Liebig University]. PMID- 10478124 TI - Please clarify misinformation about organ transplantation. PMID- 10478125 TI - Saline augmentation mammaplasty: nursing implications. AB - Saline augmentation mammaplasty has dramatically increased in popularity over the past 5 years. Maximizing immediate and long-term results requires skilled perioperative nursing intervention. PMID- 10478126 TI - The adjustable saline augmentation mammaplasty. AB - The use of an expander/implant for augmentation mammaplasty allows for increased patient satisfaction in the postoperative period. Surgical time may be shortened as final volume determination is not an intraoperative concern, and secondary surgery for certain complications such as asymmetry, wrinkling, and capsular contracture can often be avoided. PMID- 10478128 TI - Care of microvascular surgical instruments. AB - Reconstructive microsurgery, a specific category of surgery that uses revascularized tissue to repair functional or cosmetic defects, requires specialized training and continued competence of the surgeon, assistant and surgical team. Proper care of microsurgical instruments is paramount to the success of the surgery. Areas of importance such as microsurgical instrumentation, suture, operating microscope, and producing an atraumatic anastomoses will be explored indepth. PMID- 10478127 TI - Breast restoration decision making. AB - Women diagnosed with breast cancer have many physiological and psychological adjustments to make following mastectomy. The present study described in this article sought to determine the decision-making patterns used by two groups of women. Nurses need to be well informed and professionally confident to provide the individualized care and support deserved by women throughout the breast cancer trajectory. A major outcome of such professional care is confident women who understand their disease, treatments, and methods to manage stresses as they occur. PMID- 10478129 TI - Incorporating music into the surgical environment. AB - Responsibility for patient comfort has always been the concern of the nurse. Conscious sedation and or local anesthesia are commonly becoming the primary method of anesthesia for a variety of outpatient plastic surgical procedures. These surgical methods require the nurse to become aware of nonchemical adjuncts that can assist in providing a safe comfortable atmosphere for the patients. Furthermore, many patients now request nonchemical adjuncts for anxiolysis and sedation as well as for analgesia in the surgical suite. This article will provide the nurse with a basic overview of one such alternative, music, including a brief historical review, relevant research, and guidelines for use of music with the operative client. PMID- 10478130 TI - Communication skills: the art of listening. PMID- 10478131 TI - Spirituality and prayer research: a select annotated bibliography. AB - As the world has become more complex, so too have our ethical conceptualizations about it. In the 1970s, ethical theories and principle-based bioethics dominated. Then clinicians and scholars began to experience the limitations of these two approaches when used alone. In the 1980s, women's voices began to be heard through both feminist ethics and the ethic of care. In addition, virtue ethics and casuistry again gained recognition. During the 1990s and as we are about to enter the 21st century, ethics has expanded to capture the concepts of narrative ethics and spirituality. This select annotated bibliography focuses on spirituality and on prayer research. PMID- 10478132 TI - Thymoma classification. A siren's song of simplicity. PMID- 10478133 TI - Thymoma classification: my opinion. PMID- 10478134 TI - Thymoma classification. The ride of the valkyries? PMID- 10478135 TI - Post fine-needle aspiration histologic alterations of thyroid revisited. PMID- 10478136 TI - Demonstration of distinct antigenic profiles of small B-cell lymphomas by paraffin section immunohistochemistry. AB - The immunoperoxidase technique was used with antibodies against B-cell-associated antigens, including CD20, CD79a, CD10, CD23, CD43, cyclin D1, bcl-2, and kappa and lambda immunoglobulin light chains on formalin-fixed and B5-fixed tissue sections of follicular, small lymphocytic, mantle cell, and marginal zone lymphomas. Results obtained with paraffin section immunohistochemistry for CD20, CD10, CD23, and kappa and lambda light chains were compared with results obtained with flow cytometry or frozen section immunohistochemistry. Cells in all of the lymphoma types were positive for CD20 and CD79a. The antigenic profiles of the B cell lymphomas demonstrated in paraffin sections were lymphoma type distinctive. Intrafollicular lymphocytes in follicular lymphomas were positive for CD10 and bcl-2. Small lymphocytic lymphomas expressed CD43 and CD23 and were negative for CD10 and cyclin D1. Mantle cell lymphomas characteristically expressed CD43 and cyclin D1 and were negative for CD23 and CD10. Marginal zone lymphomas were negative for CD23, CD10, and cyclin D1. All of the antibodies performed better in B5-fixed tissues, but formalin-fixed tissue immunophenotypes were always similar to those obtained on the B5-fixed tissue. These results were possible using well fixed tissue, various antigen retrieval strategies, paraffin section reactive primary antibodies, and sensitive detection systems. Paraffin section immunohistochemistry on sections of routinely fixed tissue can be used similarly to flow cytometry and frozen section immunohistochemistry when classifying the lymphomas of small B lymphocytes. PMID- 10478138 TI - Autopsy findings in umbilical cord blood transplant recipients. AB - Human umbilical cord blood stem cells have been used to reconstitute hematopoiesis in patients with malignant and nonmalignant diseases. The immunologic immaturity of cord blood cells confers peculiar characteristics to these hematopoietic precursors. Autopsy reports from January 1, 1988, through June 30, 1998, were searched for patients who had received an umbilical cord blood transplant (UCBT). Thirty-two patients (19 male, 13 female) were identified with a mean age at autopsy of 13.0 years with a range from 1 to 52 years. Most patients (24) underwent UCBT for treatment of a malignant neoplasm, while the remainder were treated for immunodeficiencies (4), Lesch-Nyhan syndrome (2), Hurler syndrome (1), and Diamond-Blackfan syndrome (1). Sixteen patients had at least 1 infectious complication, and 8 patients had multiple infections. Organisms included mycoses (7 patients), viruses (8 patients), bacteria (3 patients), and Toxoplasma (2 patients). Hemorrhagic complications, such as intra alveolar hemorrhage and gastrointestinal tract hemorrhage, were found in 24 patients. Other frequent findings at autopsy included diffuse alveolar damage (15 patients), hepatic veno-occlusive disease (11 patients), and acute or chronic graft-vs-host disease (9 patients). Patients who have received UCBT represent a unique population of immunosuppressed patients. Infectious and hemorrhagic complications frequently are encountered at autopsy, and pathologists performing autopsies on these patients should be alert to unusual pathogens. PMID- 10478137 TI - Syphilitic lymphadenopathy. Histology and human immunodeficiency virus status. AB - Few reports on syphilitic lymphadenopathy have appeared in 20 years, and none have compared findings in patients with and without human immunodeficiency virus (HIV) infection, despite the recent epidemic spread of syphilis and HIV. Twelve cases of syphilitic lymphadenopathy were studied and grouped according to HIV status. Patients were 21 to 62 years old (median, 29 years); 7 were men, 5 were women. Biopsy sites were cervical (7 cases), inguinal (4), and axillary (1) lymph nodes. All patients had evidence of syphilis. Rapid plasma reagin titers ranged from 1:32 to 1:512. Treponemal hemagglutination was positive in all cases tested. Spirochetes were found with Steiner staining in 2 cases. HIV testing was positive in 4, negative in 2, and unknown in 6 cases. Lymph nodes were enlarged and often fragmented due to capsular fibrosis and chronic inflammation, with focal obliteration of the subcapsular sinus. Follicular and interfollicular hyperplasia was seen in all cases and was usually marked, with prominent vascular proliferation, plasma cells, immuno-blasts, histiocytes, and occasional neutrophils. Follicle lysis and granulomas suggestive of unconfirmed toxoplasmosis were each seen in 1 case, and Kaposi sarcoma in 2, all in HIV positive patients. Lymphoplasmacytic infiltration was marked, especially in interfollicular areas, with peri-vascular plasma cell cuffing in all cases and obliterative endarteritis in about half (7 of 12, 56%). Immunostaining for CD45RO (UCHL-1), CD20 (L26), kappa, lambda, and CD68 (Kp-1) revealed a mixed population of T cells, polyclonal B cells, and interfollicular histiocytes. Distribution of T and B cells (immunoarchitecture) was essentially normal and similar in all cases, regardless of HIV status. Syphilis produces essentially identical findings in lymph nodes in both HIV-positive and HIV-negative patients. The morphologic findings described should prompt evaluation for infection with Treponema pallidum and, in light of the current epidemic, HIV. PMID- 10478139 TI - A novel, rapid, multiparametric approach for flow cytometric analysis of intranuclear terminal deoxynucleotidyl transferase. AB - This report describes a novel, reliable, and simplified approach for determination of intranuclear terminal deoxynucleotidyl transferase (TdT) expression. This approach utilizes standard permeabilization/fixation solutions to reliably detect intracellular antigens with minimal alterations in the light scatter properties of the analyzed cells. In contrast to other described methods, fluorescein isothiocyanate-conjugated anti-TdT antibody is added to previously analyzed and permeabilized cells after a leukemic cell population has been identified using characteristic surface staining patterns. The method eliminates the need for additional sample preparation or cumbersome permeablization steps and can easily be incorporated into any clinical laboratory's existing flow cytometry panels. Sixty-eight cases were analyzed with this method, including 31 acute myelogenous leukemias, 30 acute lymphoblastic leukemias, and 7 chronic lymphoproliferative disorders. To confirm the validity of the method, parallel immunoperoxidase staining and microscopic evaluation of cytocentrifuge test sample preparations were performed. Statistical analysis of the results reveals the method to be highly sensitive and specific, demonstrating exact correlation to the microscopic method. The ease and expeditiousness of this new procedure allows TdT testing to be routinely incorporated into the immunophenotyping repertoire of a busy clinical laboratory. In addition, the method should be readily adaptable to analyze a variety of other clinically relevant intranuclear and intracytoplasmic antigens. PMID- 10478140 TI - Biomarkers of liver regeneration allow early prediction of hepatic recovery after acute necrosis. AB - Acute toxic hepatic necrosis is common and may be fatal. Predicting clinical outcome may be aided by following serum markers that could indicate recovery or may signify massive (substantial) destruction of functional liver mass. Previously, in a published case of chloroform poisoning, we serially assayed serum biomarkers of hepatocellular necrosis (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase) and markers of hepatocellular regeneration (alpha-fetoprotein, retinol-binding protein, gamma glutamyl transferase, and des-gamma-carboxyprothrombin). We noted a decline in necrotic markers and a synchronous elevation in regenerative markers, which could be suggestive of a favorable outcome in similar cases. We now report 6 Amanita mushroom poisonings with favorable outcome and 2 fatal acetaminophen poisonings in which the same markers were observed. Our results further support our hypothesis that a sustained decline in serum markers of hepatocyte necrosis with a concurrent elevation in regenerative markers could aid in prediction of favorable outcome in patients with acute liver injury. PMID- 10478141 TI - Evaluation of four automated methods for determination of whole blood cyclosporine concentrations. AB - Cyclosporine is a widely used and potent immunosuppressant drug with a narrow therapeutic index. Therefore, cyclosporine concentrations should be monitored closely. Various automated immunologic methods for cyclosporine whole blood determinations are available. Two new methods, fluorescence polarization immunoassay (FPIA) for the AxSYM by Abbott Laboratories, Chicago, IL, and the cloned enzyme donor immunoassay (CEDIA) by Boehringer Mannheim, Mannheim, Germany, have been introduced. In addition, Dade Behring improved its enzyme multiplied immunoassay (EMIT) assay. The present study evaluated all 3 new methods in comparison with high-performance liquid chromatography (HPLC) and the FPIA for the TDx analyzer. We measured whole blood cyclosporine concentrations of 179 samples obtained from 127 patients after kidney, bone marrow, heart-lung, and liver transplantation. All 4 automated immunologic methods can be used for routine measurement of cyclosporine whole blood concentrations. Disadvantages, such as higher cross-reactivity (Abbott TDx, CEDIA) or a limited linearity range (EMIT), are accompanied by advantages, such as a high precision (Abbott TDx) or an easy sample handling procedure (CEDIA). Information presented in this article should help to find the most adequate cyclosporine method for each medical laboratory. PMID- 10478142 TI - BsmI polymorphism of the vitamin D receptor gene in hyperparathyroid or hypoparathyroid dialysis patients. AB - Since bone mineral density may be influenced by the polymorphisms of the vitamin D receptor (VDR) gene, we studied whether VDR genotypes might drive the progression toward hyperparathyroidism or hypoparathyroidism in patients with end stage renal disease. On the basis of their parathyroid hormone (PTH) levels, we divided 99 patients undergoing dialysis into 2 groups: 56 patients with hypoparathyroidism (PTH < 104 pg/mL [< 11 pmol/L]) and 43 with hyperparathyroidism (PTH > 261 pg/mL [> 27.5 pmol/L]). The BB polymorphism was more frequent in patients with hypoparathyroidism (34%) than in patients with hyperparathyroidism (16%), but the difference did not reach statistical significance. Patients with the B allele and BB genotype had a significantly lower dialytic age and serum PTH and alkaline phosphatase levels than patients with the b allele and bb genotype. These results suggest that in end-stage renal disease, the BB genotype may mark a higher risk of developing hypoparathyroidism and diminished bone turnover. PMID- 10478143 TI - Distribution of carcinoma in radical prostatectomy specimens in the era of serum prostate-specific antigen testing. Implications for delivery of localized therapy. AB - Prior studies documenting the distribution of cancer in radical prostatectomy specimens were based on specimens resected before the advent of prostate-specific antigen (PSA) testing and were not specifically aimed at defining the incidence of cancer in precise regions of the gland. If localized therapy, such as brachytherapy or conformal radiation, could be minimized in areas of low cancer incidence, such as the anterior prostate, morbidity including injury to adjacent normal tissue might also be minimized. We examined 82 recent prostatectomy specimens and noted all areas of cancer in each gland. Distribution of involvement was evaluated with respect to preoperative serum PSA level, clinical stage, and pathologic features. Prostate cancer was present posteriorly in 78 of 82 cases and anteriorly in 53 of 82 cases. The presence of anterior cancer was strongly associated with elevated PSA level (P = .003), total cancer volume (P = .0002), and extraprostatic extension (P = .006), but was not associated with age, Gleason grade, or clinical stage. We conclude that the incidence of anterior prostate cancer is high. Its presence is strongly correlated with other variables, and thus strategies for selecting patients for limited localized therapy based on preoperative PSA and tumor volume estimates may be warranted. PMID- 10478144 TI - Syndecan-1 expression is induced in the stroma of infiltrating breast carcinoma. AB - Loss of expression of the heparan sulfate proteoglycan syndecan-1 leads to reduced cell adhesion, increased invasive potential, and dysregulated growth of mammary epithelial cells in vitro. We compared syndecan-1 expression in malignant and nonmalignant breast tissues using immunohisto-chemistry with monoclonal antibody B-B4. Staining for syndecan-1 is greatly diminished on malignant cells within infiltrating ductal carcinomas (n = 20) as compared with ductal epithelium of both normal breast (n = 14) and stromal-epithelial neoplasms (n = 10), which exhibit extensive basolateral epithelial staining. Surprisingly, comparison of malignant and nonmalignant breast tissue also reveals a striking difference in expression of syndecan-1 within the stromal compartment. In infiltrating ductal carcinomas, strong staining for syndecan-1 is present both within the connective tissue and on stromal cell surfaces, whereas syndecan-1 expression is absent in the stroma of both normal breast and stromal-epithelial neoplasms. Because syndecan-1 interacts with heparin-binding growth factors such as FGF-2, accumulation of syndecan-1 within the tumor stroma may contribute to the extensive angiogenesis and stromal proliferation characteristic of infiltrating breast carcinoma. Moreover, the induction of syndecan-1 within the stroma, coupled with the loss of syndecan-1 on malignant cells, suggests that changes in syndecan-1 expression are critical in promoting the metastatic phenotype of infiltrating ductal carcinoma of the breast. PMID- 10478145 TI - Subclassifying atypical squamous cells in Thin-Prep cervical cytology correlates with detection of high-risk human papillomavirus DNA. AB - Recent studies have proposed subclassifying ASCUS into "favor reactive" (ASFR), "not otherwise specified" (ASNOS), and "favor squamous intraepithelial lesion (SIL)" (ASFS). This study explored the reproducibility of these diagnoses with Thin-Prep cytology and their association with high-risk human papillomavirus DNA (HRHPV). Three pathologists and 1 cytotechnologist with 2 to 25 years of experience reviewed 144 Thin-Prep (Cytyc, Boxborough, MA) specimens previously diagnosed as normal, ASFR, ASNOS, ASFS, and SIL. Interobserver reproducibility was computed with the kappa statistic. The original laboratory diagnosis was compared with the presence of HRHPV types. Interobserver reproducibility for a normal or SIL diagnosis was very good (kappa = .68 and .63). Reproducibility for ASFR, ASNOS, and ASFS ranged from poor to fair (kappa = .21, .19, and .32). In a weighted analysis, kappa values for ASFR/ASNOS and ASFS/SIL were .36 and .62, respectively. HRHPV-positivity for preparations originally diagnosed as N, ASFR, ASNOS, ASFS, and SIL were 5.7%, 8.8%, 17.4%, 47.8%, and 54.5%, respectively. The difference in index of HRHPV for either N or ASFR and ASFS or SIL was significant (P < .001). Reproducibility for ASCUS is generally poor, but better reproducibility is obtained by combining ASFS with SIL and, to a lesser degree, ASNOS with ASFR. ASFS and SIL confer a similar index of HRHPV and merit similar management. ASFR may be managed with cytologic follow-up; but this may depend upon the individual laboratory. HPV testing, in conjunction with cytologic and biopsy follow-up, appears useful for estimating the significance of ASCUS subgroups in laboratory practice. PMID- 10478146 TI - Prognostic factors in T1 NO MO adenocarcinomas and bronchioloalveolar carcinomas of the lung. AB - There are few prognostic factors for patients with T1 N0 M0 pulmonary conventional and bronchioloalveolar adenocarcinomas (BACs), despite a 25% to 35% failure rate. To identify prognostic factors related to disease-free survival, we retrospectively studied the histologic features of 218 cases of T1 NO MO adenocarcinomas. The mean overall follow-up was 5.9 years, and the 5-year disease free survival was 72%; 148 patients (67.9%) were disease-free, and in 57 (26.1%), nonpulmonary metastases developed. Features significantly associated with decreased 5-year disease-free survival were larger tumor size, increasing central fibrosis, most common and highest nuclear grade, lymphatic vascular space invasion, and more than 50% tumor necrosis. Patients with lymphatic vascular space invasion had a 35% 5-year metastases-free survival. A tumor size of 2 to 3 cm, lymphatic vascular space invasion, highest nuclear grade 3, and increased central fibrosis were associated with metastases. Lymphatic vascular space invasion had the strongest odds ratio of 5.4. These histologic features can stratify patients with T1 N0 M0 neoplasms who have an increased risk of metastases. Future studies are needed to address the usefulness of adjuvant therapy for patients with neoplasms that display these negative factors. PMID- 10478147 TI - Immunochemical microanalysis of amyloid proteins in fine-needle aspirates of abdominal fat. AB - Diagnosis of systemic amyloidosis by fine-needle aspiration biopsy of abdominal fat is well established. A complete diagnosis now must include determination of the chemical type of amyloid. Microextracts of amyloid proteins from 11 Congo red positive aspirate samples were analyzed with immunochemical methods. There was correspondence of the results obtained by immunohistologic and Western blotting analyses in 3 of 4 specimens with kappa light chain amyloid, 5 of 6 with lambda amyloid, and 1 with amyloid A. The method provides rapid and reliable diagnostic information necessary for classification of the chemical type of amyloid required for initiation of specific modes of therapy, with little discomfort to the patient. PMID- 10478148 TI - Comparison of capillary zone and immunosubtraction with agarose gel and immunofixation electrophoresis for detecting and identifying monoclonal gammopathies. AB - Capillary zone electrophoresis (CZE) and immuno-subtraction electrophoresis (ISE) were evaluated for ability to detect and immunotype monoclonal proteins, compared with agarose gel electrophoresis (AGE) and immunofixation electrophoresis (IFE), respectively. Six hundred seventeen serum samples were analyzed with CZE and AGE to determine sensitivity and specificity in detecting IFE-confirmed monoclonal gammopathies. Both techniques detected all monoclonal spikes due to IgM (n = 8), IgG (n = 38), and free light chains (n = 3). Agarose gel electrophoresis, however, detected only 11 of 14 (79%) IgA monoclonal spikes detected with CZE. In a second study, 78 serum samples, 48 of which had a monoclonal gammopathy confirmed with IFE, were evaluated with ISE. Only 60% to 75% of the monoclonal gammopathies were correctly immunotyped with ISE by 4 readers blinded to the IFE immunotype. Thus CZE was more sensitive than AGE in detecting low concentrations of monoclonal proteins, but ISE is less accurate than IFE in determining the immunotype of the monoclonal gammopathy. PMID- 10478149 TI - Serodiagnosis of hepatitis C virus. Effect of new evaluation of cutoff values for enzyme-linked immunosorbent assay in Brazilian patients. AB - With the goal of reducing false-positive results in enzyme-linked immunosorbent assay (ELISA) serodiagnosis of hepatitis C virus in clinical practice, a study was undertaken to establish better cutoff values. We examined 277 serum samples from patients with hepatitis (non-A, non-B; B; autoimmune); subjects with antinuclear antibodies or rheumatoid factor, anticytomegalovirus or Epstein-Barr virus IgG or IgM antibodies, or parasitic disease (Chagas disease, leishmaniasis); and healthy volunteers. Concordant positive results in 2 different immunoblot assays in 250 samples were taken as indicative of true positive, and when negative, of absence of infection. Reactivity in 3 ELISA tests were evaluated for the manufacturer recommended cutoff (CO) and for 2CO, 3CO, and 4CO; and corresponding sensitivity and specificity were calculated for single or combined pairs of ELISA tests. Although CO is adequate for blood bank screening, because it provides maximal sensitivity, the frequently observed false-positive results could be significantly reduced by increasing the cutoff value to 2CO, with no significant loss in sensitivity either in relation to pairs of immunoenzymatic tests or to a single ELISA. PMID- 10478150 TI - More biopsies per block. PMID- 10478151 TI - Deaths, death notices, eulogies and obituaries. PMID- 10478152 TI - Media coverage of health stories often inaccurate, MDs report. PMID- 10478153 TI - Looking for an alternative. PMID- 10478154 TI - Considering the cost of CABG. PMID- 10478155 TI - Home is where the health care is. PMID- 10478156 TI - A straw-man argument? PMID- 10478157 TI - Smoking out the economics of tobacco use. PMID- 10478158 TI - Whiplash cultures. PMID- 10478159 TI - Which physicians make home visits and why? A survey. AB - BACKGROUND: Recent changes in the North American health care system and certain demographic factors have led to increases in home care services. Little information is available to identify the strategies that could facilitate this transformation in medical practice and ensure that such changes respond adequately to patients' needs. As a first step, the authors attempted to identify the major factors influencing physicians' home care practices in the Quebec City area. METHODS: A self-administered questionnaire was sent by mail to all 696 general practitioners working in the Quebec City area. The questionnaire was intended to gather information on physicians' personal and professional characteristics, as well as their home care practice (practice volume, characteristics of both clients and home visits, and methods of patient assessment and follow-up). RESULTS: A total of 487 physicians (70.0%) responded to the questionnaire, 283 (58.1%) of whom reported making home visits. Of these, 119 (42.0%) made fewer than 5 home visits per week, and 88 (31.1%) dedicated 3 hours or less each week to this activity. Physicians in private practice made more home visits than their counterparts in family medicine units and CLSCs (centres locaux des services communautaires [community centres for social and health services]) (mean 11.5 v. 5.8 visits per week), although the 2 groups reported spending about the same amount of time on this type of work (mean 5.6 v. 5.0 hours per week). The proportion of visits to patients in residential facilities or other private residences was greater for private practitioners than for physicians from family medicine units and CLSCs (29.7% v. 18.9% of visits), as were the proportions of visits made at the patient's request (28.0% v. 14.2% of visits) and resulting from an acute condition (21.4% v. 16.0% of visits). The proportion of physicians making home visits at the request of a CLSC was greater for those in family medicine units and CLSCs than for those in private practice (44.0% v. 11.3% of physicians), as was the proportion of physicians making home visits at the request of a colleague (18.0% v. 4.5%) or at the request of hospitals (30.0% v. 6.8%). Physicians in family medicine units and CLSCs did more follow-ups at a frequency of less than once per month than private practitioners (50.9% v. 37.1% of patients), and they treated a greater proportion of patients with cognitive disorders (17.2% v. 12.6% of patients) and palliative care needs (13.7% v. 8.6% of patients). Private practitioners made less use of CLSC resources to assess home patients or follow them. Male private practitioners made more home visits than their female counterparts (mean 12.8 v. 8.3 per week), although they spent an almost equal amount of time on this activity (mean 5.7 v. 5.2 hours per week). INTERPRETATION: These results suggest that practice patterns for home care vary according to the physician's practice setting and sex. Because of foreseeable increases in the numbers of patients needing home care, further research is required to evaluate how physicians' practices can be adapted to patients' needs in this area. PMID- 10478160 TI - Regional variations in the use of home care services in Ontario, 1993/95. AB - BACKGROUND: Although regional variations in the use of many health care services have been reported, little attention has been devoted to home care practices. Given the dramatic shift in care settings from hospitals to private homes, it is important to determine the extent to which home care practices vary by geographic region. METHODS: Data from the Canadian Institute for Health Information and the Ontario Home Care Administration System database were used to assess regional variations in rates of home care use following inpatient care and same-day surgery for the fiscal years 1993, 1994 and 1995. Various measures of regional variation were employed. RESULTS: Of the 2,870,695 inpatient separations and 1,803,307 same-day surgery separations during the study period, 359,972 and 64,541, respectively, were followed by home care. The rate of home care use per 100 separations was 12.5 for inpatients and 3.6 for same-day surgery patients. There was a a 3.5-fold regional variation in the rates of home care use following inpatient care and a 7-fold variation in rates of use following same-day surgery. Additional home care funding to attain calculated target rates was estimated to be $48.9 million (30% of expenditures for patients recently discharged from hospital over the study period). For a 20% increase in service provision it was estimated that an additional injection of $42.2 million is required. INTERPRETATION: The wide regional variations in rates of home care use highlight the importance of modifying home care funding to ensure that all residents of Ontario have equal access to services. To achieve this our estimates suggest that a substantial increase in home care funding is warranted. PMID- 10478161 TI - Going the distance: the influence of practice location on the Ontario Maternal Serum Screening Program. AB - BACKGROUND: The Ontario Maternal Serum Screening (MSS) Program was introduced by the Ontario Ministry of Health as a province-wide pilot project in 1993. The objective of this study was to determine the influence of practice location on Ontario health care providers' use of and opinions regarding MSS, access to follow-up services and recommendations about the program. METHODS: A questionnaire was mailed to a random sample of 2000 family physicians, all 565 obstetricians and all 62 registered midwives in Ontario between November 1994 and March 1995. RESULTS: Among providers who were eligible (those providing antenatal care or attending births) the response rates were 91.4% (778/851), 76.0% (273/359) and 78.0% (46/59) respectively. Fewer respondents in the Northwest region (71.4%) and in rural areas (81.9%) stated that they routinely offer MSS to all pregnant women in their practices compared with respondents in other regions (84.4%-91.5%) and urban centres (90.1%). Fewer respondents in the northern regions (Northeast 49.2%, Northwest 25.0%) than in the Central East region (includes Toronto) (76.6%) felt that follow-up services were readily available. Respondents in the northern regions had less favourable opinions of MSS than those in the other regions in terms of its complexity, cost, the time involved in counselling and the high false-positive rate. More respondents in the Central East region (64.6%) and in urban centres (52.9%) recommended not changing the MSS program than did those in the Northwest (7.1%) and rural areas (39.8%). After provider characteristics were controlled for in a logistic regression analysis, practice location was not the most important factor. Instead, the model showed that respondents who cared for 50 or more pregnant women in the previous year were more likely to offer MSS routinely (OR 2.00, 95% CI 1.21-3.27) and that those who felt that patient characteristics affect the offering of MSS (OR 0.42, 95% CI 0.26-0.67) or that follow-up services were not readily available (OR 0.33, 95% CI 0.20-0.55) were less likely to offer it. INTERPRETATION: Health care providers in northern and rural Ontario were less likely to offer MSS routinely than those in other regions and were more likely to recommend changing or eliminating the program. Providers' concerns about the social and cultural sensitivity of MSS and the availability of follow-up services affected use. PMID- 10478162 TI - Confidential prescriber feedback and education to improve antibiotic use in primary care: a controlled trial. AB - BACKGROUND: Antibiotics are a medication class for which inappropriate prescribing is frequently described. We sought to assess the effectiveness of a mailed intervention combining confidential prescribing feedback with targeted educational bulletins in increasing the use of less expensive, first-line antibiotics by practising physicians. METHODS: The participants were 251 randomly selected primary care physicians from southern Ontario who consented to participate (135 in the feedback group and 116 in the control group). Prescribing data were obtained from the claims database of the Ontario Drug Benefit program, which covers all Ontarians over age 65 years for drugs selected from a minimally restrictive formulary. Confidentially prepared profiles of antibiotic prescriptions coupled with guidelines-based educational bulletins were mailed to the intervention group every 2 months for 6 months. The control group received no intervention until after completion of the study. The main outcome measures were change from baseline in physician's median antibiotic cost and proportion of episodes of care in which a prespecified first-line antibiotic was used first. RESULTS: The median prescription cost of about $11 remained constant in the feedback group but rose in the control group (change of $0.05 v. $3.37, p < 0.002). First-line drug use increased in the feedback group but decreased in the control group (change of 2.6% v. -1.7%, p < 0.01). In a mailed survey of 100 feedback recipients (response rate 76%), 82% indicated that they would participate readily in another, similar program. INTERPRETATION: A simple program of confidential feedback and educational materials blunted cost increases, increased the use of first-line antibiotics and was highly acceptable to Ontario primary care physicians. PMID- 10478163 TI - Assessing the quality of newspaper medical advice columns for elderly readers. AB - BACKGROUND: Medical advice columns in newspapers can provide a valuable service by educating the general public about important health-related issues. However, these columns may be harmful if the advice or information given in them is incomplete, inappropriate or misleading. The objective of this study was to assess the safety and appropriateness of advice given to elderly readers of newspapers in medical advice columns. METHODS: Medical advice columns published in Canadian newspapers in 1995 were identified from a CD-ROM database. The articles that were selected were published in English and contained medical advice pertinent to elderly people about topics that could be found in a textbook of geriatric medicine. Fifty articles, randomly selected from the 109 articles that met these criteria, were independently assessed by 5 geriatricians. A scoring system was used to rate the ability to determine to which population the article applied, how well fact was distinguished from opinion, the degree to which critical issues were addressed, the safety and the appropriateness of the advice. When the kappa statistic for inter-rater agreement was 0.74 or less, a 2 stage Delphi process was used in an attempt to reach consensus. RESULTS: Agreement (kappa > 0.74) was eventually achieved for 232 (92.8%) of the 250 ratings. In 4 (8%) of the articles there was a high probability that the advice given could be applied to the wrong patient population; in 7 (14%) there was a high probability that opinion might be interpreted as fact; and in 11 (22%) the major critical issues were not identified. Of greatest concern, however, the advice in 25 (50%) of the articles was judged to be inappropriate, and in 14 (28%) advice may have been dangerous and potentially life-threatening. INTERPRETATION: Although medical advice columns have the potential to improve the health of elderly readers, a significant percentage of these articles contain inappropriate or even potentially dangerous advice. PMID- 10478164 TI - Who's afraid of the newspaper advice column? PMID- 10478165 TI - Voting ourselves rights: a critique of the Canadian Medical Association Charter for Physicians. PMID- 10478166 TI - Why physicians need a charter. PMID- 10478167 TI - Tuberculosis: 9. Treatment. PMID- 10478168 TI - Elevated levels of serum creatinine: recommendations for management and referral. AB - BACKGROUND: The potential benefits of earlier referral to a nephrologist of patients with elevated levels of serum creatinine include identifying and treating reversible causes of renal failure, slowing the rate of decline associated with progressive renal insufficiency, managing the coexisting conditions associated with chronic renal failure and facilitating efficient entry into dialysis programs for all patients who might benefit. METHODS: A subcommittee of the Canadian Society of Nephrology, which included representatives from family practice and internal medicine, conducted a MEDLINE search for the period 1966 to 1998 using the key words referral and consultation, dialysis, hemodialysis, peritoneal dialysis, renal replacement therapy and kidney diseases. Where published evidence was lacking, conclusions were reached by consensus. GUIDELINES: Earlier referral to nephrologists of patients with elevated creatinine levels is expected to lead to better health care outcomes and lower costs for both the patients and the health care system. All patients with newly discovered renal insufficiency (as evidenced by serum creatinine elevated to a level above the upper limit of the normal range of that laboratory, adjusted for age and height in children) must undergo investigations to determine the potential reversibility of disease, to evaluate the prognosis and to optimize planning of care. All patients with an established, progressive increase in serum creatinine level should be followed with a nephrologist. Adequate preparation for dialysis or transplantation (or both) requires at least 12 months of relatively frequent contact with a renal care team. Nephrologists should provide consultation in a timely manner for any patient with an elevated serum creatinine level. In addition, they should provide advice about what aspects of the condition require particularly urgent or emergency assessment. SPONSORS: This clinical practice guideline has been endorsed by the Canadian Society of Nephrology and the College of Family Physicians of Canada. Meeting, teleconference and travel expenses of the Referral Guideline Subcommittee were covered by The Momentum Program, a collaboration between Baxter Corp. and Janssen Ortho Inc. However, the authors are solely responsible for the editorial content of this article. PMID- 10478170 TI - There's a new sheriff at Tunney's Pasture. PMID- 10478169 TI - Leak of abortion information creates turmoil at Foothills. PMID- 10478171 TI - Alberta MDs line up against "antiphysician" billing restrictions. PMID- 10478172 TI - Consumers rush to buy wares at Canada's travelling "health show". PMID- 10478173 TI - Canadian Medical Association charter for physicians. PMID- 10478174 TI - [Adjuvant chemotherapy for primary breast cancer--review of 1998 St. Gallen's conference]. AB - The 6th International conference on Adjuvant Therapy of Primary Breast Cancer was held in St. Gallen, Switzerland in February 1998, and this is an overview of its guidelines. Since 1995, several specific areas have accumulated important new information and were highlighted at the 1998 conference. These include the following: genetic testing of women at risk; the availability of chemopreventive agents for high-risk and postmenopausal women; changes in diagnostic procedures with an intent to reduce axillary dissection by introducing sentinel-node biopsy; results from trials of preoperative chemotherapy; initiatives to evaluate preoperative endocrine therapies; and debate on the role of increased local control by postmastectomy radiation, its safety and effectiveness. At the conclusion of the Conference, a consensus panel of experts was asked to develop a series of guidelines and recommendations for selection of adjuvant systemic treatments in specific patient populations. The panel reviewed and modified its 1995 guidelines and recommendations based on new evidence that has emerged from clinical research. PMID- 10478175 TI - [Contribution of radiation biology to the development of radiation therapy]. AB - Results of radiation therapy for malignant tumors have steadily improved, and both radiation biology and radiation physics have contributed to this improvement. As examples of such contribution, radiobiologically-elaborated continuous hyperfractionated accelerated radiotherapy (CHART) has been proven to be superior to conventional radiotherapy against non-small cell lung cancer, and a hypoxic cell sensitizer nimorazole has been proven to be effective against pharyngeal and supraglottic laryngeal cancers. Based on laboratory studies, a combination of chemotherapy and radiotherapy has been shown to be superior to radiotherapy alone in many cancers. Radiation biology has also provided important fundamental data for clinical applications of heavy ion and proton beam therapy. In the future, useful predictive assays and radioprotectors are also expected to be developed. Radiation biology should continue to contribute to the further development of clinical radiation therapy. PMID- 10478176 TI - [Current status and future prospects of 3D treatment planning in radiation therapy, focusing on IMRT]. AB - The ultimate goal of radiation therapy is to confine a high dose to the target area while sparing the surrounding normal structures in order to increase the delivered dose and decrease the likelihood of organ injuries. In Japan, the rotational conformal technique, which is a combination of gantry rotation and dynamic movement of multi-leaf collimators (MLC), has been widely used as a standard method for high-precision radiation therapy. The non-coplanar technique in which radiation beams are given in three dimensions has the advantage of dose concentration as well as organ sparing. In intensity modulated radiotherapy (IMRT), an uneven intensity map within a beam is generated with various methods such as "sliding window technique" and "stop and shoot technique". Several intensity modulated beams are combined to create arbitrary dose distribution including concave distribution. Although a substantial number of proton therapy facilities are planned in this country, IMRT should be considered as a competitive rival from the viewpoint of cost-benefit analysis as well as clinical effectiveness. PMID- 10478177 TI - [Present and future in brachytherapy--from the discovery of radium to the 21st century]. AB - Brachytherapy can deliver high doses of radiation to a tumor with only low doses to the normal tissue. Brachytherapy can be classified as intracavitary, intraluminar and interstitial radiotherapy. It can be also divided into three groups according to dose rate: low (LDR), medium (MDR) and high (HDR) dose rates. In recent years, HDR remotely controlled afterloading systems are widespread in Japan. HDR brachytherapy has solved the problem of radiation exposure for medical staff, and patients need not be isolated in highly sealed rooms. Local control rates of T1 and T2 tongue cancer treated with LDR interstitial radiation using 226Ra and 192Ir were 80% and 67%. A phase III trial of HDR versus LDR interstitial brachytherapy for early tongue cancer revealed the same local control rates between the two groups. For uterine cervix cancer, the cause specific survival rates of patients treated with HDR intracavitary brachytherapy were almost the same as those treated with LDR. HDR brachytherapy can be applied against recurrent tumors. Almost half of recurrent tumors can be controlled with HDR treatment. Brachytherapy is widely used for prostate cancer in the USA. LDR brachytherapy using 125I seeds is used for prostate cancer. In Japan, 125I seeds can not be used because of the regulation of radioisotopes, so we treat prostate cancer patients with HDR brachytherapy. The two-year biochemical NED rate is 83%. Brachytherapy has a long history of nearly 100 years. In recent years, the development of an HDR remotely controlled afterloading system and treatment planning system allows us to make a precise treatment plan and a uniform dose distribution. In the next century, HDR-brachytherapy will continue to play an important role in the field of radiotherapy. PMID- 10478178 TI - [Progress of nuclear oncology]. AB - Radionuclides used for cancer patients are produced by cyclotron or nuclear reactor. Thyroid cancer patients with lung and/or bone metastasis have been successfully treated with 131I for more than 50 years. But for the diagnosis of cancer patients, 99mTc is superior to 131I, and many 99mTc labeled compounds have been developed, such as for the diagnosis of bone metastasis, liver and kidney diseases and so on. Recently PET studies using 18F-labeled FDG have been widely performed and excellent results are reported in many cancer patients such as lung, colorectal, malignant lymphoma, malignant melanoma and so on. Radionuclides with beta emitter are suitable for the therapy of various cancers. 131I labeled anti-CD-20 monoclonal antibodies and 131I-MIBG are used for the therapy of lymphoma and pheochromocytoma, respectively. For the palliation of metastatic bone pain, 89Sr, 153Sm or 186Re is promising, because these radionuclides can be administered on an outpatients basis. PMID- 10478179 TI - [Development of radiotherapy--history and future prospects]. AB - There have been two ways in the development of radiotherapy. One is physical development, in which efforts have been made to improve the dose concentration on the tumor while avoiding normal tissue, and the other is biological efforts to increase the radiation sensitivity of tumors. Computer medicine has improved oncological diagnoses and radiation planning, which in turn has made high quality radiotherapy possible. Radiotherapy is superior with regard to organ preservation in the treatment cancer. Elderly patients with cancer should be treated with minimally invasive organ-preserving therapy, and thus radiotherapy has been and will be an important tool for cancer treatment in the past, present and into the next century. PMID- 10478180 TI - [Clinical effect of idarubicin on malignant lymphoma]. AB - The anti-tumor efficacy and safety of idarubicin (IDR) were investigated among patients in whom the standard therapy for malignant lymphoma had failed or who had suffered a recurrence. IDR was administered at a dose of 12 mg/m2 or 15 mg/m2 by a single i.v. bolus, and this dosage regimen was repeated every 3 weeks. Among total of 21 evaluable patients, there were 3 CRs and 1 PR, for an overall response rate of 19.0% (4/21). All patients who attained CR were cases which had recurred after receiving combination chemotherapy including doxorubicin. Adverse reactions were bone marrow suppression, including leucopenia (90.5%), neutropenia (95.2%), and thrombocytopenia (47.6%); gastrointestinal symptoms such as nausea/vomiting (42.9%) and stomatitis (9.5%); alopecia (23.8%) and fever (19.0%). These results indicate that IDR is effective against refractory malignant lymphoma. PMID- 10478181 TI - [Clinical experiences of implantable vascular access device, "Vital Port" for arterial infusion chemotherapy]. AB - We had the opportunity to use "Vital Port", a subcutaneous implantable vascular access port which was developed in the U.S. and has been used clinically in a multicenter study for clinical evaluation. To prevent the result from varying by facility, standardized criteria were made. The access port was implanted in 31 patients, and then intra-arterial infusion chemotherapy was performed. The follow up period was 4 weeks. No complications were observed in any of the cases. Intra arterial infusion chemotherapy was carried out without any problem. This port is lightweight and has good biocompatibility, and the clinical results were evaluated highly. PMID- 10478182 TI - [Comparative study of histopathological effects of preoperative chemotherapy using UFT and in vitro MTT assay of colonoscopy specimens from patients with colorectal cancer]. AB - In previous studies, we demonstrated that the histopathological effects of 5-FU on colorectal cancer corresponded with the clinical effects. We measured the chemosensitivity of colorectal cancer to 5-FU by MTT assay using specimens from colonoscopy, and compared this with the histopathological effects. Forty-five patients with histologically proven colorectal cancer were enlisted for this study. Patients first underwent colonoscopy, and MTT assays were performed. Afterwards, preoperative UFT chemotherapy at a dose of 600 mg/day was given over 10 days before the operation. The histopathological effects of the resected tumors in response to 5-FU were estimated. Seven of 37 patients were judged sensitive to 5-FU by MTT assay. The response rate was 18.9%. On the other hand, 30 of 37 were evaluated as insensitive. Five of 7 patients with sensitivity to 5 FU were also evaluated as sensitive according to the histopathological effects. The sensitivity true positive rate was 71.4%. Twenty-nine patients without sensitivity to 5-FU were also evaluated as insensitive according to the histopathological effects. The specificity true negative rate was 96.7%, and the accuracy rate was 91.9%. The chemosensitivity of colorectal cancer to 5-FU, determined by MTT assay, was nearly consistent with the histopathological effects, so it may also be expected to correspond with the clinical effects. PMID- 10478184 TI - [Evaluation of amrubicin with a 5 day administration schedule in a mouse model]. AB - It was reported that amrubicin hydrochloride (9-aminoanthracycline SM-5887), showed a higher therapeutic activity than doxorubicin against human tumor xenografts implanted into nude mice with a single treatment schedule. In order to find a more effective treatment schedule, the efficacy, toxicity and pharmacokinetic properties with a 5 consecutive day treatment schedule were investigated. The total amount of the maximum tolerated dose and tumor growth inhibiting activity with a 5 day schedule was found to be higher than with a single administration. High levels of amrubicinol, the active metabolite of amrubicin, was detected in the tumor tissue. It was thus assumed that the improved efficacy with the 5-day schedule resulted from the high accumulation of amrubicinol. Bone marrow suppression at the MTD with the 5 day schedule was severer than with a single dose, but recovery was rapid, similar to that following a single dose. In conclusion, it was demonstrated that a 5 day treatment schedule was more effective than a single administration. PMID- 10478183 TI - [Clinical evaluation of granisetron (BRL43694) in nausea and vomiting induced by anticancer drugs--pediatrics]. AB - Granisetron has been used widely for the prevention and treatment of nausea and vomiting associated with anticancer drugs in adult patients with cancer. This multi-center open study was conducted to study the efficacy, safety and usefulness of granisetron in children with cancer. Among 166 evaluable patients, the efficacy rate (percentage of "remarkably effective" or "effective") was 84.9% and the usefulness rate (percentage of "extremely useful" or "useful") was 87.3%. No serious adverse effects were observed. As granisetron 40 micrograms/kg had an excellent antiemetic effect and a high degree of safety against nausea and vomiting associated with anticancer drugs, it was shown to be useful for children with cancer. PMID- 10478185 TI - [Evaluation of antitumor activity of etoposide administered orally for 21 consecutive days against human uterine cancer subcutaneous and/or orthotopic xenografts in nude mice]. AB - The antitumor activity of etoposide (ETP) against human uterine cancer cell lines were investigated in vitro and in vivo. The cytotoxic activity of ETP against HeLa S3, a human cervical cancer cell line, depended on exposure time. The survival rate with 24 h prolonged exposure was reduced to about 1/200 that with 6 h exposure. The time dependency of antitumor activity of ETP against HeLa S3 subcutaneously transplanted in nude mice was studied. The effect of 21 or 28 consecutive days oral administration was greater than that of 5 or 14 consecutive days. Furthermore, a longer administration schedule was less toxic. The antitumor activity of ETP administered orally for 21 consecutive days was compared with that of CDDP, CPT-11 and 5'-DFUR using both human uterine cancer cell lines (TCO 1, SIHA, UCC08JCK) transplanted subcutaneously in nude mice and human uterine cancer cell lines (HeLa S3, UCC08JCK) transplanted into the uterus of nude mice. ETP showed the same antitumor activity as CPT-11 and 5'-DFUR against TCO-1 and UCC08JCK, human uterine cancer cell lines transplanted subcutaneously in nude mice. ETP also showed anticancer activity against two cell lines transplanted into the uterus. The growth inhibition caused by ETP administered orally at 50 mg/kg against HeLa S3 transplanted subcutaneously was 36.7% while that against the same cell line transplanted into the uterus was 58.5%. 5'-DFUR also showed the same antitumor activity as ETP. These results suggest that long term oral administration of ETP is clinically useful for cervical cancer patients. PMID- 10478186 TI - [UFT-E granule compliance in postoperative adjuvant chemotherapy]. AB - UFT-E granules were administered as postoperative adjuvant chemotherapy to patients who had undergone surgery for gastric cancer, colorectal cancer or breast cancer. After treatment for one year, the dose conditions were investigated. The subjects were patients under 75 years of age in whom malignant tumors had been confirmed histologically, who had undergone curative resections, had no marked complications, and from whom personal informed consent had been obtained. As a rule, UFT-E granules were administered orally 450 mg (t.i.d.)/day continuously for one year beginning two weeks after surgery. The dose rate was studied from patient records and the tegafur blood concentrations on the 1st, 2nd, 6th, 8th and 12th month. The appearance of complications or clinical lab test abnormalities was also checked. A total of 19 cases were compliant among the 5 gastric cancer, 10 colorectal cancer, and 7 breast cancer patients. The mean administration period was 459 days (29-879 days), and the mean completion rate was 92.5%. The complications in 4 cases (21.1%) were relatively mild. A comparison of the prescribed dosage and patient records revealed a mean dose rate of 86.3%. From these findings, long-term administration of UFT-E granules with mild side effects is considered feasible. However, to achieve high compliance, it is considered necessary to gain a clear picture of dose conditions from patients records and other sources. PMID- 10478188 TI - [A case of adriamycin and methotrexate-resistant recurrent breast cancer treated with doxifluridine and mitomycin C]. AB - A 49-year-old woman was diagnosed with local recurrence and cervical lymph node and bone metastases 55 months after surgery for breast cancer. She was treated with goserelin acetate and tamoxifen but the disease was assessed as progressive after 8 months. Five courses of CMF therapy were performed but lung, pleural and mediastinal lymph node metastases were detected. Then, five courses of CAF therapy were carried out, but a contralateral breast metastasis was detected and the patient complained of shortness of breath. The CAF therapy was assessed as PD. We attempted administration of doxifluridine (5'-DFUR) and mitomycin C (MMC) on an outpatient basis. After 6 months, no progressive disease was detected and she was relieved of her shortness of breath. The combination therapy was assessed as long NC. Combination therapy with 5'-DFUR and MMC is thus a useful treatment for adriamycin- and methotrexate-resistant breast cancer, especially in terms of quality of life. PMID- 10478187 TI - [Multi-drug resistant breast cancer responding to chemotherapy with docetaxel (taxotere: TXT)]. AB - The patient was a fifty-five year old female who had stage IVb advanced breast cancer with hypoxia due to bilateral pulmonary metastases and lymphangitis. The cancer was adriamycin (ADM) and multi-drug resistant. We administered docetaxel (taxotere: TXT) 60 mg/m2 every 3 weeks. After 2 courses, the pulmonary metastases had become remarkably reduced in size and hypoxia was reduced and performance status (PS) improved. Major toxic defects were grade-4 neutropenia and hair loss. The patient could be discharged from the hospital without O2 inhalation and enjoyed a better quality of life (QOL). This chemotherapy is thought to be effective against ADM and multi-drug resistant breast cancer. PMID- 10478189 TI - [A breast cancer patient with docetaxel-resistant pleural recurrence with remarkable response to doxifluridine and cyclophosphamide]. AB - A 47-year-old female underwent mastectomy after neo-adjuvant chemotherapy including an anthracycline agent for advanced breast cancer. She developed a pleural metastasis on 16 months later, which was refractory to intrathoracic cisplatinum administration and intravenous Docetaxel therapy. Brain metastases were also found. We therefore treated her with whole brain radiation therapy and oral chemotherapy with 5'-DFUR and cyclophosphamide. This combination therapy produced a marked decrease in each metastasis. The adverse effects were not remarkable. This regimen may play an important role not only from the standpoint of its effectiveness against tumor growth but also the quality of life of the patient. PMID- 10478190 TI - [Chemoradiation therapy for advanced esophageal cancers--report of 3 cases]. AB - Three cases of prominent-type advanced esophageal cancer were treated with chemoradiation therapy using a 5-FU analog and low dose CDDP. All cases showed a complete response after the treatment. Only mild bone marrow suppression was found in one case. This protocol will be applied for patients with prominent-type advanced esophageal cancers, especially high risk patients. PMID- 10478191 TI - [Usefulness of combination therapy with expandable metallic stent replacement and FLEP chemotherapy (consisting of 5-FU, leucovorin, etoposide and cisplatin) for advanced esophageal cancer invading the bronchus: a case report]. AB - A 63-year-old male with an esophageal cancer invading the bronchus was treated with radiation therapy (70 Gy) from November 1995, resulting in the disappearance of the lesion. He was later readmitted due to dysphagia from the recurrence of the esophageal cancer. We diagnosed, by endoscopic and chest CT scan findings, that the esophageal cancer was invading the right bronchus. We inserted an expandable metallic stent endoscopically, and the patient could eat well. We performed two courses of FLEP chemotherapy (consisting of 5-FU, leucovorin, etoposide and cisplatin) from November 12, 1997. He then suffered from an esophagobronchial fistula, but was cured by a covered expandable metallic stent replacement. No esophageal stenoses or recurrences were seen endoscopically and he could eat well on December 7, 1998. The standard modality of treatment for patients with advanced esophageal cancer invading adjacent structures is not yet established, and the prognosis for such patients remains quite poor. We performed combination therapy with an expandable metallic stent replacement and FLEP chemotherapy, and improved the patient's quality of life. This form of therapy was thus proved useful for patients with esophageal cancer invading adjacent structures. PMID- 10478193 TI - [Mechanisms of cell adhesion and migration]. AB - In order to understand the molecular mechanisms underlying cancer metastasis, it is important to clarify the mechanisms of cell adhesion and cell migration. When epithelial cells start to migrate, cell-cell junctions are first disrupted. During migration, membrane protrusions, such as lamellipodia and filopodia, are observed externally at the cell front, and retraction at the cell rear. In addition, dynamic reorganization of the actin cytoskeleton, such as disassembly and reassembly of stress fibers at cell-cell and cell-matrix junctions and membrane protrusions, is observed internally. Our experiments have demonstrated that the Rho and Rab family small G proteins coordinately regulate cell adhesion and migration of cultured MDCK cells. The Rho family consists of the Rho, Rac, and Cdc 42 subfamilies. The Rho subfamily regulates stress fiber formation, and integrin-based cell matrix adhesion. Furthermore, the Rac and Cdc 42 subfamilies regulate lamellipodia and filopodia formation, respectively, as well as cadherin based cell-cell adhesion. The detailed modes of action of these small G proteins remain to be clarified; however, it is known that these proteins regulate cell adhesion and migration through reorganization of the actin cytoskeleton. The Rab family has over thirty members. We have found that some Rab family members are involved in HGF- or phorbol ester-induced endocytosis and exocytosis (recycling) of adhesion molecules such as integrin and cadherin. Endocytosis and exocytosis of these adhesion molecules are accompanied by disassembly and reassembly of the actin cytoskeleton, respectively, in a well coordinated manner. Thus, we propose the cooperative roles of the Rho and Rab families in cell adhesion and migration. PMID- 10478192 TI - [A case of local recurrence of rectal cancer responding to local intraarterial infusion therapy]. AB - A 42-year-old male developed pain in the right gluteal region due to local recurrence after curative resection of advanced lower rectal cancer. Radiotherapy (60 Gy) was performed, but satisfactory results were not obtained. Therefore, a reservoir was placed lowing cannulation of the internal iliac artery. The chemotherapy, in addition to intravenous administration of low dose CDDP (20 mg), included local intraarterial infusion therapy with 5-FU (1,500 mg/5 hour) once per week. After 10 courses of this chemotherapy (total dose: CDDP, 200 mg; 5-FU, 15,000 mg), the pain decreased, and the tumor size was reduced without side effects, improving the patient's QOL. At present, multidisciplinary treatments including such chemotherapy and radiotherapy is performed for local recurrence of rectal cancer, but adequate results are often not obtained. Local intraarterial infusion chemotherapy via the internal iliac artery accompanied by changes in blood flow can be safely performed on an outpatient basis, and appears to be effective for local recurrence of rectal cancer. PMID- 10478194 TI - Progress toward a mandatory post-graduate year for dentistry. Introduction. PMID- 10478195 TI - The post-graduate year: lineages, opportunities, dilemmas, and public priorities. PMID- 10478196 TI - Postdoctoral education in dentistry: preparing dental practitioners to meet the oral health needs of America in the 21st century. AB - There has been increasing interest in the organization and accreditation of Postdoctoral General Dentistry Programs (PGD). In addition, numerous national organizations have called for increases in the number of first postdoctoral year (PGY-1) positions and programs. At the same time there has been a movement to incorporate concepts of competency-based education into dental education programs in order to stress the outcomes of education rather then the process. These movements have coincided with an increased recognition that dental education will be affected by the changing demographics of our population, the emerging trends in health care delivery and financing, and the need for an increase in the number of primary care providers in dentistry, trained at an advanced level, who are capable of caring for an increasingly socially diverse and medically complex population in our country in the next century. This paper reviews the history of postdoctoral education programs in dentistry and medicine with a focus on PGD education, describes the changing health care environment in which future dental professionals will practice, and relates the dental postdoctoral experience to that in medicine. A strategy is presented for the dental profession to prepare dental practitioners with the competencies needed for the future and to create enough training opportunities to prepare these practitioners to care for the oral health needs of the nation. This proposal calls for a "National Consensus Development Conference on the Future of Postdoctoral Primary Care Education in Dentistry". This conference would define the strategies necessary to prepare dental practitioners with the competencies needed for the future and develop approaches to create enough training opportunities to prepare these practitioners to care for the oral health needs of the nation. PMID- 10478197 TI - Trends in postdoctoral dental education. PMID- 10478198 TI - A national system to support a mandated PGY-1 year: how to get there from here. Post-graduate year. PMID- 10478199 TI - Public policy consideration in adopting a mandatory PGY-1 year. Post-graduate year. PMID- 10478200 TI - The predoctoral curriculum in an era of required postgraduate dental education; or if only it were true. PMID- 10478201 TI - [Diagnostic imaging in the study of heart aging. Is the "senile heart" a fact?]. PMID- 10478202 TI - [Arthroscopic reconstruction of the posterior cruciate ligament. Magnetic resonance study for bone tunnel positioning and comparison with clinical results]. AB - INTRODUCTION: The success of arthroscopic cruciate ligament reconstruction depends on several factors, such as patient selection, correct surgical execution, and postoperative rehabilitation. Technical considerations include graft choice, positioning, fixation, intercondylar notch enlargement, and new ligament tensioning. Graft acceptance is effected by all these factors. Tunnel position is of great importance both for biomechanical reasons and optimal function of the new ligament, and to avoid stress, friction, abnormal strain, and/or damage to the reconstructed ligament. Many orthopedic and radiographic literature studies discuss the exact site of anterior cruciate ligament insertion for the best possible anatomical reconstruction. In contrast, the debate over insertional area and anatomical landmarks is open for the posterior cruciate ligament (PCL), because of the difficult execution of this type of reconstruction and the smaller number of candidates. MATERIAL AND METHODS: Fifty patients with a healthy PCL underwent MRI of the knee for other diagnostic reasons and we measured the position of PCL insertion at the tibial and femoral condyles. We also examined with MRI 20 surgical patients with a reconstructed PCL. Graft position was assessed with the same method and the results compared with physical findings of joint stability and the IKDC form score. RESULTS: Three main landmarks were found on standard axial, coronal and sagittal MR images: T1 on the tibia, and F1 and F2 on the femur. These points refer to the fibrous ligament center and designate the medial, middle and lateral portion of the tibial plateau, as well as the anterior/posterior and high/low positions on the roof of the intercondylar notch and anteromedial side of the medial condyle, respectively. According to these data, the midline position, whether slightly medial or lateral, of tibial insertion, was clinically less important. On the contrary, correct femoral tunnel positioning was found to effect subsequent joint stability and prompt rehabilitation. CONCLUSIONS: This method for MR measurement is easy and repeatable, and can be used for surgical planning and patient follow up. We found it extremely useful for the correct positioning of bone tunnels, particularly the femoral condyle, in all cases. PMID- 10478203 TI - [Chronic spondylodiscitis. Clinical aspects and imaging features]. AB - INTRODUCTION: The diagnosis of a chronic inflammatory process involving the vertebral body and disk is often very difficult because patient's history, subjective symptoms and physical findings are often unconclusive. Thus imaging techniques play a decisive role. Radiography, tomography, CT and MR have different capabilities and limitations and provide different findings in spondylodiscitis. MATERIAL AND METHODS: We observed 18 cases of spondylodiscitis in the last three years. The responsible microbe, a Staphylococcus aureus from extraosseous sites, was found in two cases at blood culture. Small cell inflammatory infiltration was confirmed with CT-guided biopsy in one case, while the other cases were diagnosed based on constant chronic back pain, feveret, moderate neutrophile leukocytosis or increased erythrosedimentation speed, plus changes in radiographic patterns following antibiotic therapy. RESULTS: Plain radiography and tomography are the techniques of choice to detect or suspect the lesion, which is then studied with CT or MRI. Clear-cut irregularities and erosions on opposing vertebral bodies, reactive bone sclerosis and reduced disk space were typical signs in our series; nine patients presented irregular cavitations(s), like bone caries, surrounded by reactive sclerosis in the body near the frontal vertebral plate. CONCLUSIONS: Together with the imaging patterns of all cases, we studied in detail three cases, relative to physical findings and diagnostic techniques. We also compared the changes in chronic spondylodiscitis with those in intraspongious herniation, intervertebral osteochondritis and severe degenerative arthritis. Bone erosions on the anterior cortical surface of the vertebral body were seen in 50% of our cases and may represent a specific sign of chronic spondylodiscitis if the finding is confirmed in further studies. PMID- 10478204 TI - [Ultrasonography contrast media Levovist and power Doppler in the study of the breast. Methodology, vascular morphology and automatic enhancement quantification with wash-in and wash-out curves]. AB - PURPOSE: To define the technique for contrast-enhanced power Doppler US studies of breast lesions and to identify possible clinical applications. MATERIAL AND METHODS: We studied 51 breast lesions detected at mammography and confirmed at cytology and/or surgical biopsy; 15 were benign lesions and 36 carcinomas, namely 14 T1ab, 29 T1c and 8 T2. We found 14 masses with regular margins, 28 with irregular margins, 1 asymmetric density, 2 architectural distortions and 6 clustered calcifications. US studies were performed with an AU5 Harmonic unit (Esaote Biomedica, Genoa, Italy) equipped with a software for online image storage, analysis and automatic quantification of US signal intensity changes after contrast agent injection, namely wash-in and wash-out contrast enhancement curves. The echocontrast agent Levovist (Schering AG, Berlin, Germany), 4.0 g preparation, was administered by i.v. injection (cubital vein) in two times at a concentration of 400 mg/mL. The first 4 mL of Levovist suspension were injected as a bolus at approximately .5 mL/s to evaluate lesion vascularization and choose the best scanning plane for wash-in and wash-out quantification. The remaining 6 mL of Levovist suspension were injected at approximately 1.0 mL/s and dedicated to wash-in and wash-out recording. RESULTS: The region of interest could not be identified in 2 of 6 calcifications. After Levovist administration, signal enhancement was seen in 36 lesions. Nonsignificant curves were obtained in 7 fibroadenomas, 1 fibrocystic mastopathy and 5 carcinomas. Pathology diagnosed an in situ component around the lesion core (true positives) in 12 carcinomas with perilesional vessels and also 3 more carcinomas with perilesional foci in situ missed at contrast-enhanced US (false negatives). The wash-in/wash-out curves of 30 carcinomas differed from those of the 6 fibroadenomas, in that the former had faster wash-in and an earlier enhancement peak, as well as longer enhancement than the latter. Moreover, fibroadenoma curves are regularly increasing, with moderate variations. As for wash-out, carcinomas exhibited three main patterns, namely a monophasic, a polyphasic and a plateau pattern. DISCUSSION: The pattern of enhancement curves in fibroadenomas is related to straight and regular vessels, while arteriovenous shunts in carcinomas cause early signal intensity peaks. Wash-out is longer in carcinomas than in fibroadenomas because the former present anarchic and tortuous vessels with slow flows. CONCLUSIONS: Levovist enhanced US is a complementary test to study known breast lesions which permits the differential diagnosis of carcinomas and fibroadenomas. Our results justify a larger clinical trial to assess the role of this technique for diagnosis, prognosis and staging purposes. PMID- 10478205 TI - [Possibility of false negative results in the diagnosis of lung neoplasms with computerized tomography]. AB - PURPOSE: To describe the radiological appearance of overlooked malignant pulmonary lesions at CT and to analyze the reasons of misdiagnosis. MATERIAL AND METHODS: Ten patients with pulmonary malignant lesions (PML) overlooked at first CT examination were selected among patients with lung cancer who were referred to our institution between 1994 and 1997. CT scans were evaluated by consensus of two chest radiologists with different experience in chest radiology, who were blinded to the final diagnosis. RESULTS: The overlooked pulmonary lesions were 5 endobronchial cancers and 5 central solitary nodules. The mean diameter of the lesions ranged 1 to 2 cm. Furthermore, 7 patients had associated pleural and/or parenchymal abnormalities. The technical quality of CT examinations was considered good in 2 cases, acceptable in 6 cases, poor in 2 cases. PML were correctly identified in 6/10 cases by the first (more experienced) radiologist and in 4/10 by the second radiologist. Four cases were considered suspect for the presence of PML by the first radiologist, 2 by the second. CONCLUSIONS: Endobronchial location of the tumors and their small size were the most frequent causes of misdiagnosis of PML at chest CT in our series. However, a systematic evaluation of CT scans can reduce the percentage of missed lesions. PMID- 10478206 TI - [Defecography in rectal wall prolapse conditions]. AB - INTRODUCTION: Pelvic floor and rectal prolapse conditions have greatly benefitted by new imaging and instrumental diagnostic approaches, and especially defecography, for both pathophysiological interpretation and differential diagnosis. We investigated the efficacy of defecography in the assessment of rectal prolapse, and in particular the role of videoproctography in diagnosing such dynamic disorders. MATERIAL AND METHODS: We selected 224 patients with rectal prolapse from a series of 1,190 consecutive subjects with evacuation disturbances examined in the last 5 years with defecography combined with videoproctography. The patients were 176 women and 48 men ranging in age 32-79 years (mean: 48). Defecography was carried out with Mahieu's technique, but we changed the filter position slightly. Sixty-seven per cent of our patients had been submitted to sigmoidoscopy, but this examination does not usually show rectal intussusception. Occult blood test in feces and double contrast barium enema were carried out in 42% and 38% of cases, respectively, to exclude any organic conditions of colon. RESULTS: Mucosal prolapse was more frequent than intussusception (71% and 34%, respectively); rectal walls went out through the anus in 12 cases of anorectal intussusception and thus caused external rectal prolapse. Rectal prolapse was associated with other anorectal alterations, such as rectocele, perineal descent and puborectalis muscle syndrome, in 96 cases. DISCUSSION AND CONCLUSIONS: The dynamic changes of ampulla are well depicted by videoproctography, which showed anorectum normalization and spontaneous reduction of invagination after intussusception. Defecography exhibited good capabilities in showing rectal wall function abnormalities. Finally, some features of videoproctography such as low radiation dose, noninvasiveness and ease of execution, make the examination acceptable to patients with anorectal disorders and for the follow-up of rectal prolapse. PMID- 10478207 TI - [Magnetic resonance with endorectal coil in the local staging of prostatic carcinoma. Comparison with histologic macrosections in 40 cases]. AB - PURPOSE: Endorectal coil MRI is widely used in the diagnostic workup of prostate cancer, but diagnostic accuracy rates reported in the literature are quite variable. We report our personal experience with endorectal coil MRI in the local staging of prostate carcinoma. MATERIAL AND METHODS: Forty consecutive patients with histologically proved prostate carcinoma were examined with endorectal coil MRI at high field strength (1.5 T). All patients underwent a sagittal T1-weighted SE location sequence (TR 400, TE 20), an axial T1-weighted SE (TR 400, TE 20), two axial T2-weighted FSE sequences (TR 3000, TE 102, ETL 8) with and without fat suppression, and a coronal T2-weighted FSE sequence (TR 3000, TE 102, ETL 8); an axial Fast Multiplanar Spoiled Gradient Recalled (FMSPGR) dynamic sequence after Gd-DTPA injection was also performed in 18 patients. MR staging of local tumor spread was done according to the current literature criteria. All patients were submitted to radical prostatectomy, and histologic macrosections on the same plane as MR images were obtained from surgical specimens. MR and histologic staging were compared to assess MR accuracy in detecting capsular infiltration, seminal vesicles and apex involvement. The diagnostic yield of Gd-DTPA was also investigated. RESULTS: MRI correctly staged 31 of 40 cases (77.5%). MR accuracy was 80% in detecting capsular infiltration (85.7% sensitivity and 73.6% specificity), 90% in seminal vesicle involvement (91.6% sensitivity, 89.2% specificity) and 72.5% in apex involvement (79.1% sensitivity, 62.5% specificity). Dynamic studies with Gd-DTPA did not improve staging accuracy in any case. DISCUSSION AND CONCLUSIONS: In agreement with most of the current literature, MRI showed moderate overall accuracy in the local staging of prostate carcinoma. Particularly, MRI had good accuracy in detecting seminal vesicle involvement but moderate sensitivity and specificity in demonstrating capsular infiltration and apex involvement. Due to its high cost, MRI should not be routinely used in prostate cancer staging but should be reserved to the patients whose clinical and serological data suggest extraprostatic tumor spread, whose preoperative demonstration could avoid noncurative surgery. PMID- 10478208 TI - [The use of dynamic Doppler color ultrasonography of the penis in the study of erectile dysfunction]. AB - INTRODUCTION: We investigated the diagnostic accuracy of dynamic color Doppler sonography (D-CDS) in men with erectile dysfunctions (ED). Terminal microcirculation alterations and their correlation with erectile response after drug testing were investigated with power Doppler energy. MATERIAL AND METHODS: 134 impotent patients were submitted to CDS during flaccidity: 8 of them (6%) had sparse hyperechoic spots inside the corpora. A standardized intracavernous injection of 10 micrograms alprostadil (PGE1) was followed by audiovisual sexual stimulation. If rigid erection was not achieved after 20 minutes, patients were randomized to redosing of either 10 micrograms PGE1 (phase 2a) or 10 micrograms PGE1 plus 1 mg phentolamine (phase 2b), with reassessment of power Doppler recordings after 20 minutes (peak systolic velocities and the resistive index, RI). RESULTS: A rigid erection (RI > .90) was achieved in 26% of patients after the first injection. As for the remaining patients, the RI was improved in 12% after phase 2a and in 34% after phase 2b (p < .05), with no differences in mean peak systolic velocities. Power Doppler showed two/three orders of distal branches of helicine arterioles with regular morphology, in most of the latter subjects. A negative correlation (r = .68, p < .001) was found between penile fibrosis and the degree of rigidity achieved after the first injection. CONCLUSIONS: Penile sonography in the flaccid state can show calcificic plaques and/or fibrosis of the corpora. Redosing of PGE1 plus phentolamine during D-CDS is a safe procedure and improves diagnostic accuracy in erectile dysfunctions, with significantly fewer non-responders than redosing of PGE1 alone. Power Doppler energy shows altered morphology of helicine arterioles otherwise missed at color Doppler and is thus recommended to make an accurate diagnosis in some men with erectile dysfunctions. PMID- 10478209 TI - [Virtual cystoscopy of bladder neoplasms. Preliminary experience]. AB - PURPOSE: We investigated the comparative sensitivity of virtual and conventional cystoscopy in patients with urinary bladder cancer. MATERIAL AND METHODS: Twelve patients (8 men and 4 women; mean age: 61 years, range: 49-73) with findings of bladder cancer at conventional cystoscopy were submitted to volumetric spiral CT of the urinary bladder. Before the examination, the urinary bladder was distended with 180-200 mL of air. CT findings were then sent to an independent workstation to generate interactive intraluminal views of the bladder. The findings of virtual cystoscopy were compared with those of conventional cystoscopy in 11 patients and with the findings of transurethral resection after urethrotomy in one patient. All lesions had pathologic confirmation. RESULTS: Eighteen (90%) of 20 masses detected on conventional cystoscopy were visualized with virtual cystoscopy. The latter allowed readers to identify 13 of 13 masses (100%) > 1 cm and only 5 of 7 (71%) < 1 cm; three 4-mm masses were missed. Although only a subjective evaluation of lesion size was possible on conventional cystoscopy, there seemed to be good agreement on mass size and site with both techniques. DISCUSSION: Cystoscopy plays a key role in the diagnosis of urinary bladder carcinoma. However cystoscopy is invasive, has a limited field of view and lacks an objective scale; moreover, it is not indicated in patients with severe urethral strictures or active vesical bleeding. In our study, virtual cystoscopy depicted all the masses > 1 cm, and a lesion in a diverticulum with a small opening. Virtual cystoscopy was also very useful in a patient with urethral stricture (who could no be submitted to conventional cystoscopy) where it showed the lesion before transurethral resection after urethrotomy. The virtual technique could also be complementary to conventional cystoscopy in evaluation of bladder base and anterior bladder neck, as well as for postchemotherapy follow up. Unfortunately virtual cystoscopy does not allow biopsy of suspicious lesions. PMID- 10478210 TI - [Nuclear medicine and rheumatology. Diagnostic and therapeutic integration]. PMID- 10478211 TI - [Management of human resources, materials, and organization processes in radioprotection]. AB - INTRODUCTION: The radiologist must learn to face daily management responsibilities and therefore he/she needs the relevant knowledge. Aside from the mechanisms of management accounting, which differ only slightly from similar analysis methods used in other centers, the managing radiologist (the person in charge) is directly responsible for planning, organizing, coordinating and controlling radiation protection, a major discipline characterizing diagnostic imaging. We will provide some practical management hints, keeping in mind that radiation protection must not be considered a simple (or annoying) technical task, but rather an extraordinary positive element for the radiologist's cultural differentiation and professional identity. MATERIAL AND METHODS: The managing radiologist can use the theory and practice of management techniques successfully applied in business, customizing them to the ethics and economics of health care. Meeting the users' needs must obviously prevail on balancing the budget from both a logical and an accounting viewpoints, since non-profit organizations are involved. In radiological practice, distinguishing the management of human from structural resources (direct funding is not presently available) permits to use internal benchmarking for the former and controlled acquisition and planned replacement of technologies in the latter, obviously after evaluation of specific indicators and according to the relevant laws and technical guidelines. RESULTS AND DISCUSSION: Managing human resources means safeguarding the patient, the operator and the population, which can be achieved or improved using benchmarking in a diagnostic imaging department. The references for best practice will be set per tabulas based on the relevant laws and (inter)national guidelines. The physical-technical and bureaucratic-administrative factors involved will be considered as process indices to evaluate the gap from normal standards. Among the different elements involved in managing structural resources, the appropriate acquisition of a piece of radiological equipment is important from both a radiation protection and an economic viewpoints. In the acquisition process, the first and the last steps (technology assessment and planned replacement, respectively) are specifically important for the radiologist and play a major role in global management. In both cases the radiologist must be able to lay out autonomous and objective working projects, also using evaluation algorithms. PMID- 10478212 TI - [Rationale and limits of echo planar magnetic resonance imaging]. AB - INTRODUCTION: The use of Echo Planar Imaging (EPI) techniques has extensively increased in the last few years. The technical improvements in new Magnetic Resonance Imaging (MRI) systems satisfy the instrumental requirements for these complex high-speed methods. The latest commercial systems are provided with standard EPI sequences. The increased data acquisition speed which characterizes EPI opens new and interesting perspectives, although the resolution is lower than the one typically achieved with conventional imaging. MATERIALS AND METHODS: The description of the data sampling process in terms of the k-space is a powerful tool for planning or understanding complex sequence diagrams like EPI. The basic concepts of the method are reported and discussed, pointing out the most noteworthy feature of the k-space, i.e., its Fourier inverse yield to the image. A generic pulse sequence has been considered to explain the sampling of k-space trajectories. Examples of ultrafast imaging techniques are subsequently considered and discussed. CONCLUSIONS: EPI improves the resolution time of MRI, which is an intrinsically slow imaging procedure. Images can be obtained in few tenth of a second so that MRI suits the short temporal range of many interesting physiological processes. Recently the main goal of functional Magnetic Resonance Imaging (fMRI) has been the study of brain activity related to local changes in brain hemodynamics. These changes cause MR signal intensity variations. In the last years, several brain activity studies have been performed by means of Positron Emission Tomography (PET) which involves the administration of radioactive substances; fMRI has the advantage that it is a noninvasive technique, although peripheral nerve stimulation effects should be considered. PMID- 10478213 TI - [Nonsecretory multiple myeloma. Differential diagnosis of a case]. PMID- 10478214 TI - [Epidermoid cyst of the mouth floor with atypical ultrasonography features. Case report]. PMID- 10478215 TI - [Arterioportal fistula caused by liver biopsy demonstrated with magnetic resonance angiography. Report of a case]. PMID- 10478217 TI - [Angioplasty with stent positioning at the level of the carotid bifurcation and dilatation with the kissing balloon technique between the origin of the internal and external carotid arteries. Report of a case]. PMID- 10478216 TI - [Retroperitoneal fibrosis caused by barium: complication of rectal perforation during double contrast enema of the colon. Report of a case]. PMID- 10478218 TI - One case of carbohydrate-deficient glycoprotein syndrome. MR findings. PMID- 10478219 TI - [Pepper syndrome. Magnetic resonance features of a case]. PMID- 10478220 TI - [Labeled aprotinin scintigraphy of amyloid. Report of a case]. PMID- 10478221 TI - Diagnosis please certificate of recognition awarded to Sergio Julio Moguillansky, MD. PMID- 10478222 TI - Radiology and Epcot celebrate new millennium with a patient education and information exhibit. PMID- 10478223 TI - Molecular imaging: exploring the next frontier. PMID- 10478224 TI - Spiral CT angiography of the pulmonary circulation. AB - Spiral computed tomographic (CT) angiography of the pulmonary circulation has emerged recently as a potential useful diagnostic method for the evaluation of the pulmonary circulation. As a minimally invasive examination, this technique is becoming widely available and has progressively replaced conventional and digital pulmonary angiography as the standard diagnostic imaging modality of the pulmonary circulation. The purpose of this review is to capture the current state of the art of the technical aspects of spiral CT angiography, with special emphasis on the post-processing techniques currently available. As CT is responsible for a considerable part of the medical radiation dose applied to the population, the current trends for dose saving will also be emphasized. With regard to the clinical applications of spiral CT angiography, its introduction into the diagnostic work-up of pulmonary embolism has considerably modified the diagnostic algorithms. Our 8-year review of the literature presented herein is expected to provide the readers with a basis for formulating an informed opinion on this topic. In addition, a large number of congenital and acquired disorders of the pulmonary circulation are also relevant to this technique, and these indications are discussed in the context of the corresponding therapeutic options. Owing to the multiple possibilities inherent to this technique, spiral CT has the potential for cost savings without reduction in image quality or diagnostic accuracy. PMID- 10478226 TI - Mucoepidermoid carcinoma of the tracheobronchial tree: radiographic and CT findings in 12 patients. AB - PURPOSE: To determine the radiographic and computed tomographic (CT) findings and clinical features of mucoepidermoid carcinoma of the tracheobronchial tree. MATERIALS AND METHODS: Chest radiographic and CT findings and clinical features of 12 histopathologically proved mucoepidermoid carcinomas in 12 consecutive patients (five male, seven female; age range, 9-72 years; mean age, 36 years) were reviewed retrospectively. RESULTS: The tumors were located at the distal trachea (n = 1) or at a main (n = 2), lobar (n = 1), or segmental (n = 8) bronchus. On chest radiographs, the tumors appeared as central masses with post obstructive pneumonia or peripheral atelectasis in four patients and as solitary pulmonary or endotracheobronchial nodules in eight. At CT, the tumors were all smoothly oval (n = 6) or lobulated (n = 6) in shape (ranging 9-40 mm in diameter), adapting to the branching features of the airways. Punctate calcification within the tumor was seen in six patients. Neither metastasis nor recurrence was seen after the surgical resection (follow-up of 8-103 months; mean, 30 months). CONCLUSION: Mucoepidermoid carcinoma of the tracheobronchial tree, usually located in a segmental bronchus, appears at CT as a smoothly oval or lobulated airway mass. It adapts to the branching features of the airways. PMID- 10478225 TI - Follicular bronchiolitis: thin-section CT and histologic findings. AB - PURPOSE: To evaluate the thin-section computed tomographic (CT) findings of follicular bronchiolitis and compare them with the histologic findings. MATERIALS AND METHODS: Thin-section CT scans obtained in 12 patients (age range, 24-77 years; mean age, 47 years) with follicular bronchiolitis proved at open lung biopsy were reviewed by two observers. Underlying conditions included rheumatoid arthritis (n = 8), mixed collagen vascular disorders (n = 2), autoimmune disorder (n = 1), and acquired immunodeficiency syndrome (n = 1). All patients had thin section CT scans (1.0-1.5-mm collimation, 11 patients; 3.0-mm collimation, one patient; high-spatial-frequency reconstruction algorithm) obtained at 10-mm intervals through the chest. RESULTS: The main CT findings included bilateral centrilobular (n = 12) and peribronchial (n = 5) nodules. All 12 patients had nodules smaller than 3 mm in diameter; six patients also had nodules 3-12 mm in diameter. Areas of ground-glass opacity were present in nine (75%) patients. Histologically, all patients had lymphoid hyperplasia along the bronchioles; eight had peribronchiolar lymphocytic infiltration. CONCLUSION: The cardinal CT feature of follicular bronchiolitis consists of small centrilobular nodules variably associated with peribronchial nodules and areas of ground-glass opacity. PMID- 10478227 TI - Electrocardiographically gated thin-section CT of the lung. AB - PURPOSE: To determine whether electrocardiographic (ECG) gating improves image quality of thin-section computed tomographic (CT) scans of the lung obtained with a subsecond CT scanner. MATERIALS AND METHODS: Thin-section CT was performed in 35 patients by using standard techniques. Three additional sections were obtained in each patient with prospective ECG gating at corresponding levels of the paracardiac lung parenchyma. Non-ECG-gated and ECG-gated sections were then rated in blinded fashion by three experienced radiologists for overall image quality, spatial resolution, and diagnostic value and for different types of respiratory and cardiac motion artifacts. RESULTS: ECG gating helped significantly reduce artifacts caused by cardiac motion (i.e., distortion of pulmonary vessels, double images, or blurring of the cardiac border) (P < .05). ECG gating did not reduce respiratory motion artifacts. In patients with heart rates of less than 76 beats per minute, ECG gating significantly improved overall image quality (P = .041). ECG gating was not perceived to increase the diagnostic value of thin-section CT scans. CONCLUSION: ECG gating improves image quality of thin-section CT scans of the lung by reducing cardiac motion artifacts that may mimic disease. It must be established whether ECG gating can help increase the diagnostic accuracy of thin section CT for the evaluation of subtle parenchymal disease. PMID- 10478228 TI - Human endothelium: endovascular biopsy and molecular analysis. AB - PURPOSE: To develop a safe and reproducible method for harvesting viable vascular endothelium to analyze gene expression at sites of vascular lesions. MATERIALS AND METHODS: Coaxial curved stainless-steel guide wires were used to obtain samples of endothelial cells from large arteries and veins in 29 patients undergoing routine endovascular procedures. Three immunocytochemical markers were used to identify cells as endothelial. Cellular viability was evaluated in terms of cell membrane integrity, energy-dependent uptake of acetylated low-density lipoprotein, and cellular response to lipopolysaccharide. Single-cell reverse transcription polymerase chain reaction (PCR) and immunocytochemistry were used to study endothelial gene expression. RESULTS: Cells with endothelial morphology and immunoreactivity for von Willebrand factor, thrombomodulin, and angiotensin converting enzyme were consistently obtained from iliac and carotid arteries and large veins (average yield [+/- standard error] from 26 iliac arteries, 262 endothelial cells +/- 45, 20%-30% of which were viable). These cells displayed induction of E-selection messenger RNA at PCR after exposure to lipopolysaccharide. Expression of vascular cell adhesion molecule 1 transcripts in endothelial cells increased with patient age (P < .01), whereas expression of intercellular adhesion molecule 1 did not. CONCLUSION: Viable endothelium can be obtained during routine angiography. Immunocytochemical and reverse transcription PCR analyses of these cells allowed determination of transcripts and proteins expressed by endothelium at sites of vascular lesions. Such information could aid in understanding mechanisms of vascular diseases and in clinical decision making. PMID- 10478229 TI - Hepatic intraarterial 131I iodized oil for treatment of hepatocellular carcinoma in patients with impeded portal venous flow. AB - PURPOSE: To evaluate the efficacy and safety of intraarterial hepatic iodine 131 iodized oil for treatment of hepatocellular carcinoma in patients with impeded portal venous flow. MATERIALS AND METHODS: Twenty-four patients (mean age, 61 years) with hepatocellular carcinoma underwent 38 courses of 131I iodized oil (one to three per patient), with a mean dose of 2,146 MBq injected into the proper hepatic artery. Hepatocellular carcinoma manifested as single nodules (n = 8; mean, 7.75 cm), multiple nodules (n = 13; mean, 5.46 cm), or a mass (n = 3) occupying more than two hepatic segments. Portal venous thrombosis was complete (n = 10), right (n = 9), left (n = 2), or multisegmental (n = 1). Two patients had hepatofugal portal flow. RESULTS: Among the 23 patients with evaluable results, response to treatment was partial in three, and disease was stable in 12 and progressive in eight. Estimated actuarial survival rates were 70%, 33%, 12%, and 6% at 3, 6, 9, and 12 months, respectively, with two patients alive at 9 and 11 months. The median survival time was 147 days. Adverse events were the early death of one patient owing to hepatic failure and transient symptomatic hepatic failure after 12 courses in nine patients. CONCLUSION: In this preliminary experience, intraarterial hepatic 131I iodized oil did not demonstrate high efficacy in the treatment of hepatocellular carcinoma in patients with portal venous thrombosis, as side effects were not rare. PMID- 10478230 TI - Multiple fluid collections: CT- or US-guided aspiration--evaluation of microbiologic results and implications for clinical practice. AB - PURPOSE: To determine if patients with multiple fluid collections need every collection aspirated and if cross-contamination is a risk if separate sterile procedures are not followed for each aspiration. MATERIALS AND METHODS: Records from 1,076 imaging-guided percutaneous aspirations and drainages over 39 months were retrospectively reviewed; 124 patients had multiple fluid collections drained, which yielded 287 aspirates. The patients were divided into two groups: those (n = 82) with multiple collections aspirated on any 1 day, and those (n = 61) with multiple collections aspirated over 10 days. Nineteen patients were included in both groups. Gram stain microscopy and culture results were compared between sequential aspirates in each patient, and their potential effects on antimicrobial therapy and theoretic risk for cross-contamination were evaluated. RESULTS: In 82 patients undergoing multiple aspirations on any 1 day, multiple microorganisms differed in 32 patients, which indicated a need for therapy change in 18 (22%) patients. In 61 patients undergoing aspiration on different days, microorganisms differed in 32 patients, which indicated a need for therapy change in 15 (25%) patients. Cross-contamination could have occurred in 28 of 93 (30%) aspirates from patients with a second or subsequent collection if separate sterile procedures had not been undertaken. CONCLUSION: When multiple fluid collections are identified, aspirates from all collections should be obtained through separate sterile procedures to ensure optimal antimicrobial coverage and avoid cross-contamination. PMID- 10478231 TI - CT fluoroscopy-guided abdominal interventions: techniques, results, and radiation exposure. AB - PURPOSE: To evaluate the benefits of computed tomographic (CT) fluoroscopy-guided interventions and assess radiation exposures incurred with CT fluoroscopy. MATERIALS AND METHODS: A 6-month period of use of CT fluoroscopy to guide abdominal biopsy procedures and catheter drainage was analyzed. Efficacy measures and needle placement and procedure room times were compared with those of the preceding 6 months during which conventional CT was used. CT fluoroscopic times and estimated radiation exposures were compared for two CT fluoroscopic methods. RESULTS: The sensitivity and negative predictive values for biopsy procedures and the success rate for needle aspiration or catheter drainages for CT fluoroscopy- 98%, 86%, and 100%, respectively--were not significantly different from those for conventional CT--95%, 80%, and 97%, respectively. Room time was not reduced significantly, but mean needle placement time for CT fluoroscopy (29 minutes; n = 95) was significantly lower than that for conventional CT (36 minutes; n = 93; P < .005). The mean patient dose index was 74 cGy. Limiting CT fluoroscopy to scanning the needle tip rather than scanning the entire needle pass significantly reduced the dose to the patient and the operator. CONCLUSION: Although CT fluoroscopy is a useful targeting technique, significant radiation exposures may result. Therefore, radiologists need to be aware of different methods of CT fluoroscopic guidance and the factors that contribute to radiation exposure. PMID- 10478232 TI - Primary musculoskeletal neoplasms: effectiveness of core-needle biopsy. AB - PURPOSE: To analyze the effectiveness of core-needle biopsy for evaluation of possible primary musculoskeletal neoplasms, which often are evaluated with open biopsy. MATERIALS AND METHODS: Core-needle biopsy was performed at a tertiary care institution in 141 patients suspected of having a mesenchymal neoplasm. In 85 patients, the lesion was in soft tissue; in 56 patients, the lesion was in bone. Eighty-nine patients had a malignant lesion, and 52 had a benign lesion. Twenty-eight patients had undergone previous surgery. RESULTS: In 105 (74%) patients, core-needle biopsy results were concordant with results from specimens subsequently obtained at surgery with respect to tumor histologic features and grade, or they provided sufficient diagnostic information to obviate surgery. In 36 (26%) patients, inaccurate core-needle biopsy results were obtained: In nine, results were imprecise about exact histologic features; in three, results were correct about histologic features but incorrect about tumor grade. In 25 (18%) patients, open biopsy was performed after core-needle biopsy. The accuracy and rate of performance of open biopsy for soft-tissue lesions were not significantly different from those for bone lesions. CONCLUSION: Percutaneous core-needle biopsy can be an effective alternative to open biopsy in the evaluation of possible mesenchymal neoplasms of either bone or soft tissue. Needle biopsy of such lesions, however, is best performed as part of a multidisciplinary team approach to tumor management. PMID- 10478233 TI - Osteoid osteoma: CT-guided percutaneous resection and follow-up in 38 patients. AB - PURPOSE: To reevaluate at medium term the results of computed tomography (CT) guided percutaneous resection of osteoid osteomas. MATERIALS AND METHODS: Thirty eight patients who had undergone treatment by means of this technique were reexamined with a mean follow-up of 3.7 years. The short- and medium-term clinical course and histologic features of the resection specimens were analyzed. RESULTS: The bone fragment could be analyzed in all cases, and the diagnosis of osteoid osteoma was confirmed in 28 patients (74%). A different diagnosis was made in six patients: mucoid cyst, subchondral arthritic geode, fibrous dysplastic lesion, focal osteochondritis, or focal chronic osteomyelitis. Cure was obtained in 32 patients (84%), whatever the cause. Complications, generally minor and transient, were observed in nine patients (24%). The most severe complications were two femoral fractures and one focal chronic osteomyelitis due to Staphylococcus aureus infection. CONCLUSION: The results of this study confirm the efficacy of percutaneous resection of osteoid osteomas and the possibility of using this method for successful treatment of other small bone lesions. PMID- 10478234 TI - Liposclerosing myxofibrous tumor: a radiologic-pathologic-distinct fibro-osseous lesion of bone with a marked predilection for the intertrochanteric region of the femur. AB - PURPOSE: To describe the characteristic features of liposclerosing myxofibrous tumor (LSMFT) of bone. MATERIALS AND METHODS: The radiographs and clinical histories of 39 patients (21 male, 18 female; mean age, 42 years; age range, 15 69 years) with histologically verified LSMFT of bone were retrospectively studied. RESULTS: LSMFT had a predilection for the proximal femur; more than 90% (30 of 33) of the femoral lesions were in the intertrochanteric region. Radiographs showed an indolent growth pattern with a well-defined and often extensively sclerotic margin. The bone contour either was normal or showed mild expansile remodeling. Mineralization within the lesion was not uncommon. Scintigrams showed mild to moderate focal tracer accumulation. Findings at computed tomography reflected those at radiography, whereas magnetic resonance imaging findings were nonspecific. Four (10%) patients had evidence of malignant transformation. CONCLUSION: The radiologic appearance of LSMFT in the intertrochanteric region of the femur is characteristic. The substantial prevalence of malignant transformation associated with LSMFT underscores the need for close observation of this lesion. PMID- 10478235 TI - Glenohumeral relationships during physiologic shoulder motion and stress testing: initial experience with open MR imaging and active imaging-plane registration. AB - PURPOSE: To test the hypotheses that open dynamic magnetic resonance (MR) imaging can (a) be used to evaluate and define normal shoulder motion in active joint motion and muscle contraction and (b) be used in conjunction with physical examination. MATERIALS AND METHODS: With an open-configuration, 0.5-T MR imaging system and active image-plane tracking, 10 shoulders were studied in five asymptomatic subjects to establish normal patterns of glenohumeral motion during abduction and adduction and internal and external rotation. Preliminary studies of physical examination during MR imaging, in which a physician examiner applied mechanical force to the humeral head, were also performed. RESULTS: During abduction and adduction and internal and external rotation maneuvers with active subjects muscle contraction, the humeral head remained precisely centered on the glenoid fossa in all asymptomatic subjects, which is in agreement with findings of previous radiographic studies. Application of force to the humeral head by an examiner was associated with as much as 6 mm of anterior translation and 13 mm of posterior translation. CONCLUSION: Dynamic MR imaging of the glenohumeral joint is possible over a wide range of physiologic motion in vertically open systems. Use of an MR tracking coil enabled accurate tracking of the anatomy of interest. These preliminary measurements of normal glenohumeral motion patterns begin to establish normal ranges of motion and constitute a necessary first step in characterizing pathologic motion in patients with common clinical problems such as instability and impingement. PMID- 10478236 TI - Case 14: intramedullary osteosclerosis. PMID- 10478237 TI - Radiologic staging in patients with endometrial cancer: a meta-analysis. AB - PURPOSE: To apply a meta-analysis to compare the utility of computed tomography (CT), ultrasonography (US), and magnetic resonance (MR) imaging in staging endometrial cancer. MATERIALS AND METHODS: Data were obtained from a MEDLINE literature search and from manual reviews of article bibliographies. Articles were selected that included results in patients with proved endometrial cancer and imaging-histopathologic correlation and that presented data that allowed calculation of contingency tables. Data for the imaging evaluation of myometrial and cervical invasion were abstracted independently by two authors. Data on year of publication, International Federation of Gynecology and Obstetrics (FIGO) stage distribution, and methodologic quality were also collected. A subgroup analysis was performed to compare contrast medium-enhanced MR imaging with nonenhanced MR imaging, US, and CT. RESULTS: Six studies met the inclusion criteria for CT; 16, for US; and 25, for MR imaging. Summary receiver operating characteristic analysis showed no significant differences in the overall performance of CT, US, and MR imaging. In the assessment of myometrial invasion, however, contrast-enhanced MR imaging performed significantly better than did nonenhanced MR imaging or US (P < .002) and demonstrated a trend toward better results, as compared with CT. The lack of data on the assessment of cervical invasion at CT or US prevented meta-analytic comparison with data obtained at MR imaging. Results were not influenced by year of publication, FIGO stage distribution, or methodologic quality. CONCLUSION: Although US, CT, or MR imaging can be used in the pretreatment evaluation of endometrial cancer, contrast enhanced MR imaging offers "one-stop" examination with the highest efficacy. PMID- 10478238 TI - Three-dimensional US of the prostate: early experience. AB - PURPOSE: To assess the feasibility of using a three-dimensional (3D) endorectal transducer at ultrasonography (US) in the prostate gland in a clinical setting. MATERIALS AND METHODS: Sixteen patients underwent 3D imaging of the prostate gland with a 3D endorectal probe following conventional two-dimensional (2D) US and prior to prostatic biopsy. Image acquisition was performed as a volume of data with nearly immediate reconstruction and simultaneous display of sectional anatomy in three orthogonal planes--sagittal plane, transverse or coronal plane, or any arbitrary oblique plane. Images were evaluated for presence of focal lesions, glandular volume, visualization of lateral and anterior portions of the gland, and extraglandular extension of tumor. RESULTS: Three-dimensional US allowed better visualization of the gland and focal lesions, especially on the coronally reconstructed images, which were judged superior to the sagittally or transversely reconstructed images for interpretation in 50% of the patients. Prostatic volumes obtained from 3D US were consistently smaller than volumes obtained from 2D US (20% difference, P = .006). Three-dimensional US was superior to 2D US in depicting tumor presence (nine of 10 right hemispheres, three of eight left hemispheres) and extraglandular extent of disease (three of five hemispheres). CONCLUSION: Three-dimensional endorectal prostatic US appears to be clinically feasible and easy to perform. Added anatomic information from the coronal plane may allow better depiction of tumors and extraglandular spread than is possible with current 2D techniques. PMID- 10478239 TI - Renal blood flow in pigs: changes depicted with contrast-enhanced harmonic US imaging during acute urinary obstruction. AB - PURPOSE: To evaluate contrast agent-enhanced harmonic ultrasonographic (US) imaging and Doppler hemodynamics during acute urinary obstruction. MATERIALS AND METHODS: In 12 piglets, the distal ureter was obstructed for 60 minutes, followed by intravenous injection of furosemide. In six piglets, ureteral pressure was further elevated to mean arterial pressure, and in six other piglets ureteral obstruction was released. Contrast-enhanced harmonic imaging was performed, and interlobar resistive index (RI) and renal blood flow were determined at baseline and during each experimental condition. A bolus injection curve was constructed by plotting mean pixel intensity versus time, and the area under this normalized curve was compared with renal blood flow values. RESULTS: Ureteral obstruction and high ureteral pressure reduced cortical renal blood flow to 88% and 66%, respectively, of baseline values. Administration of contrast agent resulted in marked homogeneous enhancement of the renal cortex. The area under the curve diminished during ureteral obstruction and correlated well with mean cortical blood flow. RI correlated well with renal perfusion pressure but poorly with changes in renal blood flow. CONCLUSION: Contrast-enhanced harmonic US imaging depicts changes in renal blood flow during acute obstruction. Interlobar RI is a good predictor of renal perfusion pressure but not of changes in renal blood flow. PMID- 10478240 TI - Effect of downstream cross-sectional area of an arterial bed on the resistive index and the early systolic acceleration. AB - PURPOSE: To evaluate the effect of the increase in total cross-sectional area of arteries as they branch beyond the main trunks on the resistive index (RI) and early systolic acceleration (ESA). MATERIALS AND METHODS: An essentially noncompliant in vitro model that used a pulsatile pump, blood-mimicking fluid, and a branching tubing network that could be configured to produce a downstream cross-sectional area one, two, four, or eight times that of the feeding vessel was used to investigate the relationship, if any, between arterial bed cross sectional area and the RI and ESA. RESULTS: The mean ESA in the branching network was inversely proportional to cross-sectional area, decreasing by approximately a factor of two for every doubling of the cross-sectional area. The mean RI in the branching network decreased with increasing cross-sectional area, but not as greatly as the ESA did; the mean RI in the bed with eight times the upstream cross-sectional area had an RI that was approximately three-fourths the upstream RI. These relationships are real, as the slopes of the plots (ESA vs cross sectional area, P = .001; RI vs cross-sectional area, P < .02) are significantly different from zero. CONCLUSION: RI and ESA decrease as a result of increasing downstream cross-sectional diameter of the arterial bed. PMID- 10478242 TI - Brachytherapy for the prevention of stenosis in a canine hemodialysis graft model: preliminary observations. AB - PURPOSE: To determine whether gamma brachytherapy can prevent in-stent stenosis in hemodialysis grafts. MATERIALS AND METHODS: Six-millimeter polytetrafluoroethylene arteriovenous grafts were created bilaterally in six dogs. After 1 month, Wallstents spanning the venous anastomosis were placed to accelerate restenosis. Gamma irradiation (12 Gy) was delivered endoluminally to one of the two grafts by using an iridium 192 source; thus, each animal served as its own control. Fistulography was performed monthly for 10 months or until graft thrombosis, with measurement of stenosis at each time point. At the conclusion of the study period, the treated area was examined histologically, and a computer model was used to calculate the volume of intimal hyperplasia. RESULTS: Delayed stent migration resulted in exclusion of one dog. In the remaining five dogs; maximum stenosis across all time intervals was less for the treated side (P < .04), and the volume of intimal hyperplasia was less for the treated side (P < .045). In one animal studied at 1 year, this trend reversed in terms of percentage stenosis but not total neointimal volume. CONCLUSION: Brachytherapy with 192Ir (gamma) delivered at the time of stent placement reduces restenosis in this hemodialysis graft model, but, depending on the parameter evaluated (stenosis vs total volume of neointima), the benefit may wane or even reverse with time. PMID- 10478241 TI - Simultaneous noninvasive determination of regional myocardial perfusion and oxygen content in rabbits: toward direct measurement of myocardial oxygen consumption at MR imaging. AB - PURPOSE: To determine whether myocardial arterial perfusion and oxygen concentration can be quantified simultaneously from the same images by using spin labeling and the blood oxygenation level-dependent (BOLD) effect with fast spin echo (SE) imaging. MATERIALS AND METHODS: A T2-weighted fast SE pulse sequence was written to image isolated, arrested, blood-perfused rabbit hearts (n = 6) at 4.7 T. Perfusion images with intensity in units of milliliters per minute per gram that covered the entire left ventricle with 0.39 x 0.39 x 3.00-mm resolution were obtained in less than 15 minutes with a 32-fold reduction in imaging time from that of a previous study. Estimates of oxygen concentration were made from the same images acquired for calculation of perfusion images. RESULTS: Estimates of regional myocardial oxygen content could be made from the perfusion images; this demonstrated the feasibility of three-dimensional calculation of regional oxygen consumption, which requires concomitant measurement of both oxygen content and flow. Fast SE imaging was shown to be as sensitive to hemoglobin desaturation as standard SE imaging. Perfusion abnormalities and oxygen deficits were easily identified and verified qualitatively with gadopentetate dimeglumine on both perfusion and BOLD images obtained after coronary arterial ligation. CONCLUSION: T2-weighted fast SE imaging combined with perfusion-sensitive spin labeling can be used to measure myocardial arterial perfusion and oxygen concentration. This provides the groundwork for calculation of regional myocardial oxygen consumption. PMID- 10478243 TI - Phases IB and II multidose trial of gadolinium texaphyrin, a radiation sensitizer detectable at MR imaging: preliminary results in brain metastases. AB - PURPOSE: To evaluate magnetic resonance (MR) imaging results after administration of gadolinium texaphyrin, a tumor-selective radiation sensitizer that is detectable at MR imaging, and to determine an appropriate intravenous dose of gadolinium texaphyrin for repeated injections during radiation therapy, the dose limiting toxicity of reiterated doses of gadolinium texaphyrin, the maximal tolerated dose, the biolocalization of gadolinium texaphyrin (as assessed at MR examinations), and the response to treatment. MATERIALS AND METHODS: Ten daily intravenous injections of gadolinium texaphyrin, each followed by whole-brain radiation therapy (total of 10 fractions, 30 Gy), were administered to patients with brain metastases in a multicenter study. At the study institution, 11 patients underwent MR imaging before and after the first injection, after the 10th injection, and 8 weeks after entry into the study. RESULTS: MR imaging revealed selective drug uptake in metastases, without enhancement of normal brain tissue. In 10 patients, tumor uptake was higher after the 10th injection than after the first injection, which indicated accumulation of gadolinium texaphyrin in metastases. One lesion was visible only after the 10th injection and not at the pretherapeutic MR examination with injection of conventional gadolinium-based contrast material. Response to treatment was defined as a reduction in the size of the metastases between the preinjection MR study and the last MR study; seven patients achieved partial remission with tumor regression exceeding 50% of the initial size, and four achieved a minor response with less than 50% tumor regression. CONCLUSION: These preliminary results indicate that gadolinium texaphyrin is tumor selective and that brain metastases can be depicted at MR imaging long after the administration of gadolinium texaphyrin. PMID- 10478244 TI - The radial bands sign. PMID- 10478246 TI - Quantitative diffusion-tensor anisotropy brain MR imaging: normative human data and anatomic analysis. AB - PURPOSE: To obtain normative human cerebral data and evaluate the anatomic information in quantitative diffusion anisotropy magnetic resonance (MR) imaging. MATERIALS AND METHODS: Quantitative diffusion anisotropy MR images were obtained in 13 healthy adults by using single-shot echo-planar MR imaging and a combination of tetrahedral and orthogonal gradient encoding (whole-brain coverage in about 1 minute). White matter (WM) anatomy was assessed at visual inspection, and values were measured in various brain regions. Different anisotropy measures, including total anisotropy (A sigma), were compared on the basis of information content, rotational invariance, and susceptibility to noise. Partial volume and noise effects were simulated. RESULTS: Anisotropy MR images depicted WM features not typically seen on conventional MR images (e.g., external capsule, thalamic substructures, basal ganglia, occipital WM, thickness of the internal capsule). Statistically significant anisotropy differences occurred across brain regions, which were reproducible within and across subjects. A sigma was highest in commissural WM and progressively lower in projection and association WM. This order paralleled that of known resistance to spread of vasogenic edema, which suggested that anisotropy may be sensitive to WM histologic structure. Gray matter (GM) A sigma data were consistent with zero anisotropy, and partial volume WM-GM effects were approximately linear. A sigma image quality could be effectively improved by means of averaging. CONCLUSION: Quantitative diffusion anisotropy images can be obtained rapidly and demonstrate subtle WM anatomy. Different histologic types of WM have significant and reproducible anisotropy differences. PMID- 10478245 TI - Motor neuron diseases: comparison of single-voxel proton MR spectroscopy of the motor cortex with MR imaging of the brain. AB - PURPOSE: To evaluate single-voxel proton magnetic resonance (MR) spectroscopy in detection of abnormality of the upper motor neuron in patients with motor neuron diseases. MATERIALS AND METHODS: In 43 of 50 patients with motor neuron disease and 14 of 14 control subjects, matching sets of MR spectra were obtained in the left and right motor cortex. The ratio of N-acetylaspartate (NAA) to creatine (Cr) was derived from peak area measurements. Mean ratios were calculated for control subjects and several patient groups, including patients with amyotrophic lateral sclerosis (ALS) or primary lateral sclerosis (PLS). MR images were evaluated for corticospinal tract hyperintensity and central sulcus dilatation. RESULTS: Mean NAA/Cr values were significantly different between control subjects and the ALS or PLS groups (P < .05). With an optimal cutoff of 2.5, NAA/Cr values were abnormal in 15 (79%) of 19 patients with ALS, 12 (67%) of 18 patients with PLS, and one (7%) of 14 control subjects. Corticospinal tract hyperintensity, central sulcus enlargement, or both were found in 43% of the ALS group, 24% of the PLS group, and 7% of the control group. CONCLUSION: NAA/Cr values determined at single-voxel proton MR spectroscopy are more sensitive than are standard findings at MR imaging in the detection of upper motor neuron disease. PMID- 10478247 TI - Human acute cerebral ischemia: detection of changes in water diffusion anisotropy by using MR imaging. AB - PURPOSE: To (a) determine the optimal choice of a scalar metric of anisotropy and (b) determine by means of magnetic resonance imaging if changes in diffusion anisotropy occurred in acute human ischemic stroke. MATERIALS AND METHODS: The full diffusion tensor over the entire brain was measured. To optimize the choice of a scalar anisotropy metric, the performances of scalar indices in simulated models and in a healthy volunteer were analyzed. The anisotropy, trace apparent diffusion coefficient (ADC), and eigenvalues of the diffusion tensor in lesions and contralateral normal brain were compared in 50 patients with stroke. RESULTS: Changes in anisotropy in patients were quantified by using fractional anisotropy because it provided the best performance in terms of contrast-to-noise ratio as a function of signal-to-noise ratio in simulations. The anisotropy of ischemic white matter decreased (P = .01). Changes in anisotropy in ischemic gray matter were not significant (P = .63). The trace ADC decreased for ischemic gray matter and white matter (P < .001). The first and second eigenvalues decreased in both ischemic gray and ischemic white matter (P < .001). The third eigenvalue decreased in ischemic gray (P = .001) and white matter (P = .03). CONCLUSION: Gray matter is mildly anisotropic in normal and early ischemic states. However, early white matter ischemia is associated with not only changes in trace ADC values but also significant changes in the anisotropy, or shape, of the water self-diffusion tensor. PMID- 10478248 TI - Brain lesions: when should fluid-attenuated inversion-recovery sequences be used in MR evaluation? AB - PURPOSE: To compare qualitatively and quantitatively the contrast of brain lesions detected with fluid-attenuated inversion-recovery (FLAIR) and intermediate-weighted sequences at magnetic resonance (MR) imaging. MATERIALS AND METHODS: In this prospective study, 47 patients suspected of having a brain lesion underwent MR imaging with FLAIR, intermediate-weighted, and T2-weighted sequences. Qualitative assessment was performed of lesion conspicuity, detection, overall image artifact, and additional clinical information. Contrast and contrast-to-noise ratio (CNR) were calculated between lesions and the normal brain or cerebrospinal fluid (CSF). RESULTS: FLAIR images were equal to intermediate-weighted images for overall lesion conspicuity and detection but were associated more often with image artifacts. Lesion-to-background contrast was significantly higher on FLAIR than on intermediate-weighted images. FLAIR images failed to demonstrate multiple sclerosis (MS) plaques located in the basal ganglia and brain stem. CONCLUSION: Although FLAIR images provided additional information in some cases, they did not have distinct advantages over intermediate-weighted images. When cases of MS are evaluated, intermediate weighted images are preferable to FLAIR images. Except in cases of MS, either FLAIR or intermediate-weighted sequences should be added to T2-weighted sequences at MR imaging. PMID- 10478249 TI - Primary and recurrent early stage laryngeal cancer: preliminary results of 2 [fluorine 18]fluoro-2-deoxy-D-glucose PET imaging. AB - PURPOSE: To evaluate the effectiveness of 2-[fluorine 18]fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) in the identification of early stage (T1 T2) primary and recurrent laryngeal cancer. MATERIALS AND METHODS: Twelve patients with T1 or T2 laryngeal cancer underwent imaging prospectively with PET. Seven patients had new disease, and five had recurrent disease. All patients underwent imaging prior to planned therapy and tissue biopsy. PET images were evaluated by using standardized uptake ratios and visual analysis. RESULTS: Histopathologic evidence of early stage cancer was documented in the 12 patients. One had a carcinoma in situ, nine had T1 tumors, and two had T2 tumors. Of the 12 patients, 10 had vocal cord tumors, one had a hypopharyngeal tumor, and one had a preepiglottic tumor. Eleven (92%) patients with early stage cancer had standardized uptake ratios indicative of malignancy (mean, 4.6; SD, 1.8; 95% CI, 1.2; range, 2.8-7.6). One had false-negative results (standardized uptake ratio = 2.3). Nine underwent CT, and results in the larynx were normal in seven and abnormal in two. CONCLUSION: FDG PET can be used to identify primary and recurrent early stage laryngeal cancer. It may be useful for follow-up after therapy. PMID- 10478250 TI - Staging non-small cell lung cancer with whole-body PET. AB - PURPOSE: To compare the accuracies of whole-body 2-[fluorine 18]fluoro-2-deoxy-D glucose (FDG) positron emission tomography (PET) and conventional imaging (thoracic computed tomography [CT], bone scintigraphy, and brain CT or magnetic resonance [MR] imaging) in staging bronchogenic carcinoma. MATERIALS AND METHODS: Within 20 months, 100 patients with newly diagnosed bronchogenic carcinoma underwent whole-body FDG PET and chest CT. Ninety of these patients underwent radionuclide bone scintigraphy, and 70 patients underwent brain CT or MR imaging. For each patient, all examinations were completed within 1 month. A radiologic stage was assigned by using PET and conventional imaging independently and was compared with the pathologic stage. The accuracy, sensitivity, specificity, and negative and positive predictive values were calculated. RESULTS: PET staging was accurate in 83 (83%) patients; conventional imaging staging was accurate in 65 (65%) patients (P < .005). Staging with mediastinal lymph nodes was correct by using PET in 67 (85%) patients and by using CT in 46 (58%) patients (P < .001). Nine (9%) patients had metastases demonstrated by using PET that were not found with conventional imaging, whereas 10 (10%) patients suspected of having metastases because of conventional imaging findings were correctly shown with PET to not have metastases. CONCLUSION: Whole-body PET was more accurate than thoracic CT, bone scintigraphy, and brain CT or MR imaging in staging bronchogenic carcinoma. PMID- 10478251 TI - Brain tumor volume measurement: comparison of manual and semiautomated methods. AB - PURPOSE: To compare the reliability of two approaches to measuring enhancing brain tumor volumes--the conventional manual trace method and a threshold-based, semiautomated computer software method. MATERIALS AND METHODS: Two operators rated contrast material-enhanced, T1-weighted axial magnetic resonance (MR) image data sets from 16 patients aged 21-71 years with high-grade gliomas. Each MR data set was rated twice by using manual tracing and twice by using the semiautomated method. The semiautomated measurement method involved a thresholding algorithm based on mixture modeling. The data collection time for each method was recorded. Reliability was measured by using inter- and intraoperator agreement indexes. RESULTS: Mean intraoperator agreement indexes (+/- SD) were 0.90 +/- 0.09 (operator 1) and 0.83 +/- 0.15 (operator 2) for the manual trace method and 0.83 +/- 0.17 (operator 1) and 0.84 +/- 0.16 (operator 2) for the semiautomated measurement method. The mean interoperator agreement was 0.85 +/- 0.14 for the manual method and 0.82 +/- 0.18 for the semiautomated method. The semiautomated method was faster than the manual trace method by an average of 4.6 minutes per patient. CONCLUSION: The semiautomated computer method of measuring tumor volume was faster than the manual trace method. Semiautomated computer approaches offer an alternative to manual tracing for measuring serial tumor volumes in patients with high-grade brain neoplasms. PMID- 10478252 TI - Improvement of radiologists' characterization of mammographic masses by using computer-aided diagnosis: an ROC study. AB - PURPOSE: To evaluate the effects of computer-aided diagnosis (CAD) on radiologists' classification of malignant and benign masses seen on mammograms. MATERIALS AND METHODS: The authors previously developed an automated computer program for estimation of the relative malignancy rating of masses. In the present study, the authors conducted observer performance experiments with receiver operating characteristic (ROC) methodology to evaluate the effects of computer estimates on radiologists' confidence ratings. Six radiologists assessed biopsy-proved masses with and without CAD. Two experiments, one with a single view and the other with two views, were conducted. The classification accuracy was quantified by using the area under the ROC curve, Az. RESULTS: For the reading of 238 images, the Az value for the computer classifier was 0.92. The radiologists' Az values ranged from 0.79 to 0.92 without CAD and improved to 0.87 0.96 with CAD. For the reading of a subset of 76 paired views, the radiologists' Az values ranged from 0.88 to 0.95 without CAD and improved to 0.93-0.97 with CAD. Improvements in the reading of the two sets of images were statistically significant (P = .022 and .007, respectively). An improved positive predictive value as a function of the false-negative fraction was predicted from the improved ROC curves. CONCLUSION: CAD may be useful for assisting radiologists in classification of masses and thereby potentially help reduce unnecessary biopsies. PMID- 10478253 TI - Local tumor recurrence following breast-conservation therapy: correlation of histopathologic findings with detection method and mammographic findings. AB - PURPOSE: To correlate histopathologic findings with detection method and mammographic appearance in primary and locally recurrent breast carcinoma after breast-conservation therapy. MATERIALS AND METHODS: Medical records and mammographic findings were retrospectively reviewed; 26 patients with 27 local recurrences after breast-conservation therapy were identified. RESULTS: Primary histopathologic findings included six in situ and 20 invasive carcinomas. Of the 27 recurrences, 19 (70%) were at or adjacent to the lumpectomy site and eight (30%) were elsewhere in the breast. All primary ductal carcinoma in situ (DCIS) cases manifested mammographically as microcalcifications and recurred as DCIS with microcalcifications. Eleven primary invasive carcinomas (10 masses, one case of microcalcifications) were detected only mammographically, three were detected only with physical examination, and six (six masses) were detected with both. Among these 20 recurrences, 14 (five masses, nine cases of microcalcifications) were detected only mammographically, one was detected only with physical examination, and five (five masses) were detected with both. Seventeen (85%) of 20 primary invasive carcinomas recurred invasively: 16 (94%) with similar histopathologic findings and eight (47%) with similar mammographic findings. CONCLUSION: In local recurrence after breast-conservation therapy for DCIS, histopathologic findings, detection method, and mammographic findings are usually similar. Histopathologic findings of primary invasive breast carcinoma and local recurrence are usually similar, but the detection method and mammographic findings vary. This is relevant to the interpretation of new clinical or mammographic findings following lumpectomy. PMID- 10478254 TI - Anterior chest wall: frequency of anatomic variations in children. AB - PURPOSE: To evaluate the frequency of anterior chest wall variations in children. MATERIALS AND METHODS: The computed tomographic (CT) images of 200 consecutive infants and children (114 boys and 86 girls; mean age, 10.5 years; age range, 3 months to 19 years) who underwent chest CT during a 20-month period were evaluated for chest wall variations. Children who had undergone chest wall surgery or were suspected of having a chest wall abnormality were excluded. The frequency of chest wall anomalies was compared with age and sex (Fisher exact test). RESULTS: The CT scans of 65 children (33%) depicted one or more variations in the anterior chest wall: titled sternum (n = 29), prominent convexity of anterior rib or costal cartilage (n = 19), prominent asymmetric costal cartilage (n = 20), well-defined paracostal subcutaneous nodule (n = 4), mild pectus excavatum (n = 4), or mild pectus carinatum (n = 4). The frequency of these findings did not vary significantly with age (P = .96) or sex (P = .36). CONCLUSION: Variations in the anterior chest wall are common, occurring in one third of children, and should be considered normal. These asymptomatic variations should not be considered alarming when palpated at physical examination. PMID- 10478255 TI - Soft-tissue venous malformations in pediatric and young adult patients: diagnosis with Doppler US. AB - PURPOSE: To describe the diagnostic features, appearance, and vascularization pattern of venous malformations (VMs) at Doppler ultrasonography (US). MATERIALS AND METHODS: Between February 1991 and May 1997, 51 soft-tissue VMs were studied with Doppler US in patients between 1 day and 21 years of age (mean age, 9 years). These VMs were located in the maxillofacial region (n = 19), trunk (n = 5), and upper (n = 10) and lower (n = 17) extremities. Twenty-three VMs had venographic confirmation, seven had only histologic confirmation, and 21 had both venographic and histologic confirmation. US was performed with 7.5- or 7-10-MHz linear transducers, a low pulse repetition frequency (mean, 1,680 Hz), and the lowest wall filter (25-50 Hz). RESULTS: At gray-scale US, VMs appeared as hypoechoic, heterogeneous lesions in 82% of cases. All lesions displayed compressibility. In eight lesions (16%), phleboliths were identified, thus confirming the diagnosis of VM. Analysis of vascular flow revealed monophasic, low-velocity flow in 40 VMs (78%), with an average flow velocity of 0.22 kHz. Biphasic flow was noted at the periphery of three lesions, which is indicative of a mixed capillary-venous malformation. The remaining eight lesions did not display any flow. CONCLUSION: In pediatric patients, Doppler US is a noninvasive, easily available, and rapid mode of investigation of vascular lesions and can help confirm the diagnosis of VM when it shows a characteristic flow pattern. PMID- 10478256 TI - Kyphomelic dysplasia: clinical and radiologic long-term follow-up of one case and review of the literature. AB - The authors describe the 17-year follow-up of the (to their knowledge) only adult and only female patient affected with kyphomelic dysplasia so far described in the literature, with assessment of the phenotypic, orthopedic, and radiologic progression of this syndrome. PMID- 10478257 TI - Adult intussusception detected at CT or MR imaging: clinical-imaging correlation. AB - PURPOSE: To determine the clinical presentation in cases of adult intussusception demonstrated at computed tomography (CT) or magnetic resonance (MR) imaging and to correlate the imaging appearance with clinical diagnosis. MATERIALS AND METHODS: Retrospective review of CT and MR images and clinical records of all patients with an intussusception demonstrated on CT or MR images from January 1, 1991, through April 30, 1998. RESULTS: Thirty-three patients had one or more intussusceptions demonstrated on CT (n = 30) or MR (n = 3) images. Twenty-nine patients had enteroenteric intussusceptions, and four had intussusceptions involving the colon. Ten patients (30%) had a neoplastic lead point, including all four of the intussusceptions involving the colon (benign mass, n = 3; malignant mass, n = 7). In 23 cases (70%), no neoplastic lead point was identified. A variety of causes were implicated in these cases, with 16 cases (48%) classified as idiopathic. Enteric intussusceptions in the nonneoplastic group were shorter in length (median, 4 vs 10.8 cm; P = .002), smaller in diameter (median, 3 vs 4 cm; P = .002), and less likely to be associated with obstruction (4.3% vs 50%; P = .02). CONCLUSION: Less than one-third of adult intussusceptions demonstrated at CT or MR imaging were caused by a neoplastic lead point. Almost half of adult cases in this series were idiopathic. PMID- 10478258 TI - Tumorous gastric varices: radiographic findings in 10 patients. AB - PURPOSE: To determine the appearance of tumorous gastric varices on double contrast barium studies and whether these varices have characteristic radiographic features. MATERIALS AND METHODS: Review of radiology files revealed 86 patients with gastric varices diagnosed during double-contrast upper gastrointestinal tract examinations. Of these 86 patients, 12 (14%) had a conglomerate mass of varices, or tumorous varices. Five of the 12 patients had proved gastric varices and five were presumed to have varices on the basis of additional diagnostic test results, clinical follow-up findings, or both. Radiographs from these 10 patients were reviewed retrospectively to determine the size, location, and morphologic features of these lesions. RESULTS: Tumorous varices had a mean size of 6.8 cm (range, 3-11 cm). They involved the posteromedial border of the gastric fundus in eight patients, the central cardiac region in one, and the anterolateral-inferior fundal border in one. Viewed in profile, the varices appeared as smooth submucosal masses with undulating contours and discrete borders. Viewed en face, the varices manifested as a conglomerate of thickened, tortuous folds that faded peripherally into the adjacent mucosa. CONCLUSION: Tumorous gastric varices manifest as remarkably similar findings on double-contrast barium studies, usually appearing en face as a conglomerate of thickened, lobulated folds and in profile as smooth, undulating, submucosal masses on the posteromedial border of the gastric fundus. It is important to be aware of the characteristic features of tumorous gastric varices on double-contrast studies so that they are not mistaken for neoplastic lesions in the stomach. PMID- 10478259 TI - Hepatocellular carcinoma and intrahepatic peripheral cholangiocarcinoma: enhancement patterns with quadruple phase helical CT--a comparative study. AB - PURPOSE: To define the hemodynamic features of hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma by using quadruple phase helical computed tomography (CT) and determine the value of this information in characterizing tumors. MATERIALS AND METHODS: Helical CT of the liver was performed in 45 patients with newly diagnosed HCC or peripheral cholangiocarcinoma. Scans were obtained before and 25 seconds, 70 seconds, and 2-6 minutes after the start of the contrast material injection. The intensity and spatial distribution of contrast material uptake were evaluated during all phases. Time-attenuation curves were established for each lesion. Relative attenuation and lesion conspicuity were assessed. A diagnostic confidence level was assigned to each lesion. RESULTS: In the majority of HCC lesions, a single, early peak of enhancement followed by a continuous decrease in tumor attenuation over time was seen. The greatest tumor conspicuity occurred during the delayed phase. In cholangiocarcinoma, tumor attenuation increased during the delayed phase. In the majority of lesions, the greatest tumor conspicuity was seen during the portal venous phase. In both tumor types, the diagnostic confidence level improved when the delayed phase was used. CONCLUSION: The variation over time in the intensity of contrast enhancement in HCC and cholangiocarcinoma differs sufficiently to make this a useful diagnostic criterion. The delayed phase is particularly important because it amplifies this difference. PMID- 10478260 TI - Abdominal MR imaging with a volumetric interpolated breath-hold examination. AB - PURPOSE: To compare a T1-weighted, three-dimensional (3D), gradient-echo (GRE) sequence for magnetic resonance (MR) imaging of the body (volumetric interpolated breath-hold examination, or VIBE) with a two-dimensional (2D) GRE breath-hold equivalent. MATERIALS AND METHODS: Twenty consecutive patients underwent 1.5-T MR imaging. The examinations included pre- and postcontrast (20 mL gadopentetate dimeglumine) fat-saturated 2D GRE breath-hold imaging and fat-saturated volumetric interpolated breath-hold imaging before, during (arterial phase), and after injection, with thin (2-mm source images) and thick (8-mm reconstruction images) sections. The three images were compared qualitatively and quantitatively (signal-to-noise ratio [SNR] and contrast-to-noise ratio [CNR]). RESULTS: Qualitatively, the 2-mm source images had poorer pancreatic edge definition on precontrast images compared with the other two data sets (P < .05). On gadolinium enhanced images, scores for clarity of pancreatic edge, number of vessels visualized, and arterial ghosting were significantly lower for the postcontrast 2D GRE images. Quantitatively, SNR measurements in the liver, aorta, and renal cortex on pre- and postcontrast images were significantly higher for the 8-mm reconstruction images than for the 2D GRE or 2-mm source images (P < .05). Aorta to-fat CNR was significantly higher on the 8-mm reconstruction images. CONCLUSION: Fat-saturated volumetric interpolated breath-hold images have quality comparable to that of conventional fat-saturated 2D GRE images. PMID- 10478261 TI - Cystic fibrosis: combined hyperpolarized 3He-enhanced and conventional proton MR imaging in the lung--preliminary observations. AB - Four patients with cystic fibrosis (CF) were examined with combined hyperpolarized helium 3-enhanced and conventional proton magnetic resonance (MR) imaging. After inhalation of the polarized 3He gas, single breath-hold, gradient echo images (resonant frequency of 3He) were obtained to depict lung ventilation. Conventional T2-weighted fast spin-echo (hydrogen) images were also obtained to depict morphologic abnormalities. 3He images were successfully and reproducibly generated that showed both morphologic abnormalities and, often more extensive, ventilation abnormalities. 3He MR imaging may provide a method for evaluating progression of pulmonary disease in patients with CF. PMID- 10478262 TI - Pulmonary perfusion: respiratory-triggered three-dimensional MR imaging with arterial spin tagging--preliminary results in healthy volunteers. AB - The authors used a spin-tagging method of magnetic resonance perfusion imaging to measure pulmonary perfusion in eight healthy volunteers with use of a respiratory triggered three-dimensional pulse sequence. The average signal intensity (SI) decrease upon arterial labeling was 24%. The perfusion SI increased by 21% after exercise (P = .02). Focal blood flow abnormalities were observed in a patient with chronic obstructive pulmonary disease. PMID- 10478263 TI - Pulmonary arterial resistance: noninvasive measurement with indexes of pulmonary flow estimated at velocity-encoded MR imaging--preliminary experience. AB - Cardiac output and pulmonary vascular resistance (PVR) were measured in 19 patients by means of catheterization of the right side of the heart. Results were compared with the cardiac output and indexes of pulmonary arterial blood flow estimated with velocity-encoded magnetic resonance (MR) imaging. Correlations were good between estimates with right-sided heart catheterization and those with velocity-encoded MR imaging. By providing accurate pulmonary arterial blood flow measurements, velocity-encoded MR imaging allowed distinction of patients with high PVR from subjects with normal PVR. PMID- 10478264 TI - Pulsed magnetization transfer imaging: evaluation of technique. AB - With use of an established physical model, numeric simulations were performed to evaluate current imaging protocols for the two primary applications of magnetization transfer: cerebral magnetic resonance angiography and neuroimaging of white matter disease. The authors found that the current technique is appropriate in the former but suboptimal in the latter. Further clinical investigations could potentially improve magnetization protocols for neuroimaging. PMID- 10478265 TI - ROC curve analysis in the evaluation of contrast-enhanced color Doppler US. PMID- 10478266 TI - Neurocardiology: the benefits of irregularity. The basics of methodology, physiology and current clinical applications. AB - Cardiovascular variability in heart rate and blood pressure can be used to quantify the autonomic modulation of cardiovascular function. Although the first studies were only published 10 to 20 years ago, there is presently a vast amount of knowledge from numerous publications that allows a better insight into the neural control mechanisms of the heart. The development of methods such as heart rate variability and baroreflex sensitivity prompted a multidisciplinary approach since many aspects were involved: mathematics, signal processing, physiology and cardiology. Therefore, it is obvious that an adequate understanding of certain basic principles is necessary before its clinical potential can be fully exploited. This current review outlines the physiological background and the fundamental methodology used in the field of neurocardiology by presenting the newer insights and developments from both fundamental and clinical sides. Recent publications indicate that neurocardiological applications in (risk stratification of) myocardial infarction, congestive heart failure and heart transplant patients can be expected to enter daily clinical practice in the next millennium. PMID- 10478267 TI - C.D. de Langen--his life and work. PMID- 10478268 TI - C.D. de Langen and nutrition research. PMID- 10478269 TI - Diet-heart research in the first part of the 20th century. PMID- 10478270 TI - On the determinants of mortality at the population level. AB - To attain a long and healthy life has always been one of the major goals of humanity. The Chinese emperors, the Egyptian pharaohs and the Aztec kings all aspired to become immortal. To try to avoid mortality and to decrease morbidity at the population level is the role of public health, taken in its broadest sense, including both preventive and curative medicine. In this review we will try to identify the major determinants of mortality. Only when we are able to identify the major causes of mortality and morbidity can public health be approached in a rational way. The emphasis will be on mortality because, in our opinion, populations with a long life expectancy also have a low age-specific morbidity. PMID- 10478271 TI - Cholesterol lowering by diet: what can be achieved? AB - The cholesterol concentration in the plasma is poorly regulated by the human organism and is therefore easily influenced by lifestyle. Indeed cholesterol levels differ between populations. An important question is how much diet can be held responsible for differences in cholesterol level between e.g. Finland and Japan at the beginning of the Seven Countries Study around 1960. This difference can partly be explained by a difference in saturated fat intake but there was probably an accumulation of both genetic cholesterol elevating factors present in the Finnish society and of cholesterol-lowering factors in the Japanese society. However, controlled trials do show a major effect of diet on cholesterol. With iso-caloric diets low in saturated fat and high in fibre a reduction in total cholesterol of 20-30% can be realized. The reduction can be larger if weight loss also occurs. Further reductions may occur when novel foods are introduced on the market e.g. margarines enriched with plant sterols that give an additional reduction in total cholesterol of about 10%. It can be concluded that diet can reduce total cholesterol levels substantially. The value of novel foods in this context has to be evaluated. PMID- 10478272 TI - Serum cholesterol in cross-cultural perspective. The Seven Countries Study. AB - C.D. de Langen hypothesized already in 1916 that overnutrition by a cholesterol rich diet is associated with hypercholesterolaemia and the occurrence of atherosclerotic complications such as coronary heart disease. It took till the nineteen fifties before the associations between diet, serum cholesterol and coronary heart disease were systematically investigated. Keys started the Seven Countries Study because he was as De Langen interested in the question whether differences in the occurrence of coronary heart disease between populations could be explained by differences in serum cholesterol and diet. The results of the Seven Countries Study showed that serum cholesterol was strongly related to coronary heart disease mortality both at the population and at the individual level. The strength of the association (relative risks) between serum cholesterol and coronary heart disease mortality was similar in different cultures. However, the absolute risks differed substantially. At a serum cholesterol level of 5.2 mmol/l the 25-year coronary heart disease mortality rate was 5 times higher in Northern Europe compared with Mediterranean Southern Europe. At the population level differences in coronary heart disease mortality rates could almost completely be explained by differences in saturated fat (an important determinant of serum cholesterol), flavonoids (strong antioxidants) and cigarette smoking (producer of pro-oxidants). It can be concluded that the relations between diet, serum cholesterol and coronary heart disease are more complex than originally thought. Not only dietary cholesterol but also different fatty acids and antioxidants play a role in the genesis of atherosclerosis. PMID- 10478273 TI - Clinical approaches to healthier diet modifications. AB - There is a strong relation between diet and risk factors for cardiovascular diseases. Generally a high fat consumption will increase the serum cholesterol level. Different fatty acids have different effects on total cholesterol, LDL and HDL lipoproteins and therefore on the risk for cardiovascular diseases. This knowledge was translated into guidelines for a healthy diet and in advices to use less fat. In reality it was not easy for physicians to reach a substantial reduction in total cholesterol by limiting the fat consumption. A recent investigation in Belgium showed that the median reduction obtained by dietary changes through general practitioners was only 3.5%. In medical education not much attention is paid to the communication of the "fat message". In addition, individual patients have frequently multiple risk factors e.g. abdominal obesity, insulin resistance, diabetes, hypertension and a sedentary lifestyle. Therefore in the clinical setting an integrated approach is needed, in which dietary advice is supported by behavioural therapy and suggestions to increase physical activity. Currently there is a debate on the preference of a low fat or a modified fat diet. A low fat diet is rich in carbohydrates and a modified fat diet is rich in mono-unsaturated fatty acids. Recent investigations in diabetic patients are in favour of a modified fat diet. This has the advantage that in practice the possibilities for realizing a low saturated fat diet are increased. PMID- 10478274 TI - Treatment and prevention of coronary heart disease by lowering serum cholesterol levels; from the pioneer work of C.D. de Langen to the third "Dutch Consensus on Cholesterol". AB - In the beginning of this century a possible relation was observed between cholesterol-rich foods, blood cholesterol levels and atherosclerosis by "pioneers" in this field as Anitschkow and De Langen. In the second half of this century a definite link was established between serum cholesterol levels and development of coronary heart disease (CHD). In angiographic studies it has recently been shown that a decrease in total cholesterol as well as in low density lipoprotein cholesterol level results in a retardation of the progression of vascular disease. Furthermore, clinical event intervention trials demonstrated that therapy with cholesterol synthesis inhibitors reduces not only cardiovascular and total mortality, but also other manifestations of CHD. These recent results prompted to revise, for the second time, the Dutch consensus text for lipid lowering therapy, with the following conclusions. Hypercholesterolaemia is treated with a low-saturated fat diet and normalisation of weight. For individuals, this might result in a reduction of the risk for myocardial infarction or death and for the population in a decrease of the mean serum cholesterol concentration and a reduction of the incidence of CHD. The indication for drug therapy is founded on the expected effectiveness to reduce the incidence of (new manifestations of) CHD, which is related to the level of the absolute risk of vascular disease. Treatment with cholesterol synthesis inhibitors must be considered in (a) patients with familial hypercholesterolaemia; (b) all patients with a history of myocardial infarction or other symptomatic vascular disease with a total cholesterol concentration above 5.0 mmol/l and a life expectancy of at least five years; (c) persons without known vascular disease with a combination of diabetes mellitus, hypertension, hypercholesterolaemia, cigarette smoking and high risk for development of CHD, rising from 25% per 10 years at the age of 40 years to 35-40% per 10 years at the age of 70 years, with a life expectancy of at least five years. If these guidelines are followed, the calculated cost-effectiveness is about Dfl. 40,000 per life year gained or less. The consensus committee judges this reasonable in comparison with other therapeutic interventions in the Netherlands. Thus by now, with regard to lipids and atherosclerosis, the definite link has been established between observational medicine and an effective treatment modality which is applicable in daily practise. PMID- 10478275 TI - Diet, cholesterol and coronary heart disease. A perspective. AB - After pioneering, scattered observations in the context of the so-called geographical pathology, collected during the first part of this century, a systematic approach into the search of causality of the association between diet and coronary heart disease has characterized the past 50 years. The possible link of eating habits leading to a diet high in calories, total fat, saturated fatty acids and cholesterol, to coronary events, through the mediation of serum cholesterol, represents the classic diet-heart theory. This theory is supported by results from ecological analyses, prospective studies, animal experiments, clinical observations, controlled trials, biochemical and nutritional studies. This traditional view of the problem has partly changed recently and a number of other dietary components in the possibly causal chain have emerged, with effects not necessarily involving serum cholesterol and lipids. The role of other nutrients in the atherosclerotic process and of precipitating factors (e.g. vitamins, anti-oxidants, fibre, sub-categories of fatty acids, phytosterols, flavonoids, etc.) has been identified but is not always fully understood. At the same time other blood lipids, other blood components and other pathways have been identified between diet and coronary outcomes. New challenges are open for future research since the association of diet with coronary disease is not as simple and linear as supposed. The main issues are the need to explain a number of still unknown mechanisms, to determine which "natural diet" carries the minimum coronary risk and whether "new" foods produced by modern technology are really needed to contrast this epidemic. PMID- 10478276 TI - [Thromboembolism--genetic and acquired risk factors]. AB - Within the last few years, the knowledge of hereditary and acquired risk factors for venous thromboembolism has increased. Antithrombin-, protein C- and protein S deficiency have been known since a long time as hereditary risk factors. Since 1993, three hitherto unknown risk factors have been described, the APC (activated protein C) resistance, hyperhomocysteinemia and a polymorphism in the 3-UT region of the prothrombin gene. These risk factors are relatively common in the normal population (in total 10-15%) and are found in 30-50% of patients with venous thromboembolism. The most important acquired risk factor for thromboembolism is the antiphospholipid antibody syndrome (APLS). The APLS is found in around 3% of patients with thromboembolism, patients with these abnormalities have a high risk for recurrency. The upper mentioned risk factors for thromboembolism do not only increase the risk for spontaneous thrombosis, but also the risk for thrombosis during typical high risk situations, such as surgery, trauma of the lower extremities, pregnancy and delivery. APC resistance and antithrombin deficiency increase the risk for development of thrombosis during oral contraceptive intake. Patients, in whom one of the upper mentioned risk factors have been diagnosed, should receive thrombosis prophylaxis during high risk situations. Not all patients with one thromboembolic event and a known risk factor are candidates for long-term oral anticoagulant treatment. Long-term oral anticoagulant treatment should be introduced after exclusion of major contraindications in patients with recurrent events, patients with a combination of risk factors and a life threatening event. PMID- 10478277 TI - [Deep vein thrombosis--diagnosis and therapy]. AB - For diagnosis of deep vein thrombosis an objective visualising method is indispensable (Duplex sonography, phlebography, MRI). The approval of several preparations of low-molecular-weight heparin made therapy easy: subcutaneous injections of fixed doses which are mainly adjusted to body-weight allow safe and effective anticoagulation without the need of special laboratory testing. Mobile patients should be kept walking with compression of their legs. Secondary prophylaxis consists of oral anticoagulation or alternatively in low-molecular weight heparin and compression stockings. PMID- 10478278 TI - [Diagnosis of pulmonary embolism]. AB - Although clinical assessment (good medical history, identifying patients at risk, recognition of common symptoms and signs, physical findings) and bed-side methods (standard laboratory tests, ECG, chest X-ray, arterial blood gas analysis, echocardiography and compression ultrasound of the lower extremities) for the diagnosis and/or exclusion of pulmonary embolism is highly insensitive and unspecific, the definition of clinical probability (pretest probability) still seems to be of outmost importance. Clinical probability guides initial medical therapy, induces further invasive and non invasive examinations (e.g. perfusion lung scan, spiral CT, gadolinium-enhanced magnetic resonance pulmonary angiography, contrast pulmonary angiography) and is still the final determinant in case of conflicting results of imaging modalities. PMID- 10478279 TI - [Transthoracic sonography in the diagnosis of pulmonary embolism]. AB - In many cases of pulmonary diseases extending up to the pleura, ultrasound helps to identify the etiology of the lesion. There are several sonomorphological criteria to differentiate peripheral pulmonary consolidations. Clinical studies and the sonographic appearance with pathologic correlation showed pulmonary infarctions in location, form and size exactly corresponding with pathological findings. Fresh reperfusable infarcts were homogenous and hypoechoic. Older infarcts were well demarcated, mainly wedge shaped. Triangular pleural based lesions, more roughly structured, were observed with a hyperechoic reflex in the center corresponding to the bronchiole: a sign of segmental involvement. The sensitivity of transthoracic ultrasound in diagnosis of pulmonary embolism was 86 to 94%, the specificity 67 to 87%, positive predictive value 55 to 92%, negative predictive value 91%, accuracy 73 to 91%. In massive central lung embolism, both fresh and old infarctions are found. An imminent larger embolism can be predicted, for instance,in a deep vein thrombosis. With one ultrasound system, we can "kill three birds with one stone": source, way and outcome of pulmonary embolism by bedside examination. PMID- 10478280 TI - [Ischemic stroke: new therapeutic possibilities]. AB - Recent randomized placebo-controlled studies have introduced new concepts in the therapy of acute ischemic stroke, such as thrombolysis either with intraarterial or intravenous administration, lowering of the level of fibrinogen or administration of aspirin. High dosis of heparin may be useful only in certain groups of patients. Stroke units proved to be effective in the acute care of stroke patients. Further advances are achieved by the introduction of new anti platelet drugs in secondary stroke prevention and by precise definitions of the criteria for carotid surgery. PMID- 10478281 TI - [Acute coronary syndromes: physiopathology and therapeutic aspects]. AB - This article discusses recent developments in the field of acute coronary syndromes including pathophysiological mechanisms as well as therapeutic strategies. A plaque disruption is caused by different stimuli in a plaque prone to rupture, i.e. a plaque with a lipid-rich core and high local concentration of inflammatory cells (T-cells, monocytes/macrophages, mast cells). These cells are capable of producing matrix degradation products and can reduce stability of a plaque. Thrombus formation, based on platelet activation and aggregation as well as fibrin formation, is the main consequence of plaque disruption. Depending on the degree of thrombus formation occlusion is followed clinically by unstable angina (subtotal occlusion) or by acute myocardial infarction (total occlusion). Accompanying vasoconstriction may further aggravate the situation. Principles of therapy are thrombus dissolution as well as prevention of new thrombus formation: main goals of thrombolytic therapy in acute myocardial infarction are a prompt (less than 3 hours), complete, and sustained (prevention of early thrombotic reocclusion) reperfusion. PMID- 10478283 TI - Early parental preoccupations and behaviors and their possible relationship to the symptoms of obsessive-compulsive disorder. AB - This study focuses on early parental preoccupations and behaviors (EPPB) surrounding the birth of a new family member. An interview instrument was developed to assess EPPB at eight months prepartum and two weeks and three months postpartum. Measures with adequate psychometric and conceptual properties were used to assess the overall level of parental preoccupations and associated actions as well as three content domains: caregiving (CARE), relationship building (RELATIONSHIP), and anxious intrusive thoughts associated with harm avoidant behaviors (AITHAB). The hypothesis that preoccupations and behaviors would peak for both parents close to the birth of the child was confirmed. Measures of EPPB were distinguished from symptoms of depression and generalized anxiety. Consistent with our a priori hypothesis the content and character of the AITHAB was found to resemble the symptoms of obsessive-compulsive disorder (OCD). The study findings suggest the potential adaptive significance of EPPB and the possible evolutionary origins of OCD. PMID- 10478285 TI - UK primary care databases lead the way. PMID- 10478284 TI - Traditional Chinese medicines: safety hazards. PMID- 10478286 TI - Cardiotoxicity of cytotoxic chemotherapy and its prevention. PMID- 10478287 TI - [Bicentennial of the death of Lazzaro Spallanzani, pioneer of cardiovascular and respiratory physiology]. PMID- 10478288 TI - [Persistent common trunk. Anatamopathological study of 25 specimen]. AB - In order to inform the pathologic features and their associated anomalies twenty five hearts with common trunk were studied with the segmental sequential system. An anatomico-embryological correlation was made to understand the pathological complex of this malformation. The results were: type I truncus (96%), infundibular ventricular septal defect (96%), displastic truncal valve (28%), ventriculo infundibular fold (92%), left coronary artery arising from the posterior wall of the truncus (75%), right coronary artery arising from their anterior wall (96%), coronary arteries arising from opposite Valsalva sinuses in the tetracuspid valves; the biventricular conection of the truncus was balanced in 60%, prevailing on the right (16%) or on the left (16%) ventricles. Among the associated anomalies there were right aortic arch, interruption of the aortic arch, anomalous origin of the left subclavian artery, vascular ring, aneurysm of the sinus of Valsalva, and absence of the left branch of the pulmonary artery. Developmentally common trunk is explained as a failure of truncoconal septation in the embryonic heart; a migration arrest of neural crest cells is implicated in the Di George syndrome. Knowledge of the anatomic features of common trunk and their associated anomalies, provides the morphological basis to interpret correctly the clinical diagnostic imagenology. PMID- 10478289 TI - [The benefit of a coronary stent in the reduction of major cardiac complications in acute coronary syndromes]. AB - To assess the incidence of in-hospital major adverse cardiac events (MACE), we analyzed 694 procedures in 613 consecutive patients during one year period. Patient population included 550 (79.2%) patients with unstable angina, 43 (6.2%) with stable angina and 101 (14.5%) with acute myocardial infarction. Elective percutaneous transluminal coronary angioplasty (PTCA) was performed in 593 (85.4%) patients, rescue PTCA in 7 (1%), and primary PTCA in 94 (13.5%). Angiographic lesion morphology was as follows: type A 30%; type B 58%; type C 12%. We compared patient population who received stent with PTCA-balloon only. Technical success was 95% and clinical success was achieved in 80% of the cases. Overall mortality was 1% in the stent group and 3% in the conventional PTCA. The incidence of MACE was 4% and 15.1% in the stent and angioplasty balloon groups respectively. We found a dramatic impact on reduction of the incidence of acute complications in the groups with stenting for unstable angina (p = 0.0001) and acute myocardial infarction (p = 0.0001). The major clinical advantage of stenting over balloon angioplasty was a lower need for repeated procedures. PMID- 10478290 TI - [Medical-surgical management of left heart valve prosthesis dysfunction due to obstruction. Eleven year's experience]. AB - We report the surgical-medical experience in left side mechanic prosthesis cardiac valve dysfunction. From January 1986 to June 1997 we included 108 consecutive patients (pts), 24 were men and 84 women, mean age 45 years, they underwent 114 surgical procedures, 104 in mitral and 10 in aortic position. RESULTS: The dysfunction was due to obstruction in fixed disc in 107 pts and intermitent in 7 pts. The pathological findings were thrombosis in 92 pts, pannus in 9 pts and both in 13 pts. The time interval between valve replacement and dysfunction was from 1 to 247 months, mean 53.8 +/- 56.7. Seventy five percent of patients received inadequate anticoagulant therapy, the mean INR was 2.06 +/- 0.79. In 76% of patients the functional class was III or IV NYHA. The diagnosis was made by clinical and echocardiographic findings in 96% of patients. The surgical procedures included valve replacement in 103, and toilette in 11 pts. The time of extracorporeal circulation in patients who died us survival patients were (159.9 +/- 108.95 vs 87.32 +/- 27.53 min) and aortic cross-clamp (64.8 +/- 20.69 vs 48.28 +/- 23.71 min) respectively (p < 0.001). The overall mortality was 12 pts (10.5%), all in functional class III or IV NYHA (p < 0.05), five patients died during surgical procedure. In conclusion the diagnosis must be established clinically as well as by echocardiography. The risk factors associated with mortality were pulmonary edema, shock, delayed surgery, surgical time prolonged and functional class III or IV NYHA. PMID- 10478291 TI - [Percutaneous mitral commissurotomy in patients with mitral valve stenosis and thrombosis of the left atrium]. AB - Mitral percutaneous valvotomy (MPV) is an effective therapeutic alternative in the treatment of patients with mitral stenosis (MS) and thrombus in the left atrium have been an absolute contraindication to this procedure. The aim of our study is to evaluate the safety of MPV in patients with MS and thrombus in the appendix of the left atrium (ALA). Between September 1996 and April 1997, we performed ten procedures of MPV with Inoue's technique in patients with MS and thrombus in ALA. Nine females and 1 male were included. Their were 41 +/- 7.6 years old. The Wilkins score in our patients were 8 +/- 1.2. All were treated with oral anticoagulants between three and twelve months but the thrombus did not resolved. With transesophagic echocardiogram and fluoroscopic control we avoided the contact between wires and Inoue's catheter with ALA. Results were positive. Mitral valve area increased from 0.8 +/- 0.2 cm2 to 1.95 +/- 0.3 cm2. There was no case of systemic embolism or others major complications during the MVP and during following one year. CONCLUSION: MPV is a safe and efficient therapeutic alternative in the treatment of selected patients with MS and thrombus in ALA. PMID- 10478292 TI - Infectious endocarditis in pacemaker endocardial leads: report of three cases. AB - Three cases of endocarditis affecting endocardial leads of permanent pacemakers are presented with a review of the literature. Vegetations were identified using transesophageal echocardiography. Infection of pacemaker leads is far less common than infection at the site of the pulse generator with greater morbidity and mortality and generally requiring surgical removal of both electrodes and power source. The most frequent infective agents are stahylococcus varieties. PMID- 10478293 TI - Interruption of the aortic arch in adults. AB - The interruption of the aortic arch is a rare cardiac congenital disease; such patients may occasionally survive to adulthood without surgery. The associated intracardiac malformations may modify survival. Our aim is to report three young adults (18-19 years of age) with interruption of the aortic arch. Two had type C, and the other had type B with subpulmonic ventricular septal defect and pulmonary valve insufficiency. We review 106 cases collected from the medical literature into 3 groups: 1) the whole group of patients; 2) patients with isolated interruption of the aortic arch, and 3) patients with interruption of the aortic arch associated with ventricular septal defect. In the whole group we found 18 cases of interruption of the aortic arch type A, and 25 cases of interruption of the aortic arch type B; 37 cases of isolated interruption of the aortic arch and 43 cases associated with ventricular septal defect. Fifty percent of the patients died before 15 days of life (0.042 years). According to the cumulative frequency graphic, only 5% of the patients survived beyond the age of 5 years. We found no information to relate patient's survival rate and anatomic type of the interruption of the aortic arch. From adolescence, the survival of the patients with interruption of the aortic arch associated to septal ventricular defect was 7%, and a 14% survival was found in patients with isolated interruption of the aortic arch. No statistical difference was found between the means of the ages of these two groups (P > 0.25). PMID- 10478294 TI - [Translumbar coronary angiography and aortography in a patient with Takayasu's arteritis]. AB - Since its introduction by Dos Santos in 1929, arterial angiography by translumbar percutaneous approach has suffered some transformations. Nowadays it has been replaced by other percutaneous approaches and it is indicated only when these routes of access have failed due to aortoiliac or subclavian arteries obstruction. This report presents a patient with Takayasu's Arteritis with severe peripheral arterial obstruction and unstable angina, who underwent coronary arteriography and aortography by translumbar approach. A review of this technique is made. PMID- 10478295 TI - [Nutritional evaluation and functional class in hospitalized cardiopathy patients]. AB - The nutritional state evaluation of any patient with heart disease must include the anthropometric measures, organic metabolic and cellular immunity test. We evaluated the nutritional state of 75 hospitalized patients with heart disease, and its correlation with New York Heart Association class and heart disease type. There was 36 patients (48%) with normal nutritional state, 24 (32%) with grade I malnutrition, 12 (16%) with grade II malnutrition, and 3 (4%) with grade III malnutrition. Of 23 patients with rheumatic valvular heart disease 83.4% have some degree of malnutrition, 37 patients with ischemic heart disease 25% was under nourished. Fifty percent of patients with hypertensive cardiopathy, 75% of the patients with cardiomyopathy and 83% of the 7 patients with other type of heart disease had some degree of malnourishment. There was a direct correlation between nutritional state and functional class, we found no patient in IV class functional with normal nutritional state, or grade I malnutrition. PMID- 10478296 TI - [Cardiology review. Molecular and genetic aspects of cardiopathies]. PMID- 10478297 TI - [New strategies for the classification and treatment of acute ischemic coronary syndromes. A proposal]. PMID- 10478298 TI - [Freud as neuropathologist, his reception in France before 1910]. AB - Even before 1910 Freud in Paris was not unknown. His relation to Charcot first and the reception of his papers by Charcot's followers (mainly Raymond, Marie, Brissaud) later represent a particular chapter of history. This is well documented by Charcot's letters to Freud (hitherto published only in French), by Freud's French articles as well as by many medical theses (Paris, Lyon, Nancy, Bordeaux) discussing the pros and cons. Two topics were focussed. First there was a neurological dispute about Little's disease, its clinical vignette, etiologies, and its spinal and cerebral localisations. Freud argued there to be a nosographic entity. Most of Charcot's disciples, however, pleaded against the idea of an entity for such different pathological forms. This was followed by a discussion about Freuds doctrine of neuroses and their sexual origin, which Freud opposed to the French theories of heredity and degeneration. Both of the discussions were without personal encounter and without a direct exchange of views between the protagonists. Which did not cause it to be less vivid and intense. This proves Freud's standing as an internationally acknowledged expert for neuropathological aspects of the child. However, starting in 1900, Freud retired from the neurological discussion and the Paris scene. The dispute about Little's disease faded away and the discussion about neuroses became occupied by Janet's writings. The neuropathologist Freud eclipsed the psychoanalyst Freud. Hence Freud's repeated complaints not to have been well received by the French. PMID- 10478299 TI - [Persecution in Nazi everyday life. The pathological aspects of extreme stress]. AB - The extensive literature on persecution during the Third Reich lacks a first person account of the psychological distress induced by public Nazi harassments in everyday life--with the only exception of Victor Klemperer's diary. His records are outstanding in their unusual authenticity. They have gained widespread publicity and have been extensively reviewed, but almost exclusively from historical or political perspectives. The following is focused on his personal view. Taking his diaries as a starting point three subjects are treated: It is first discussed what kind of persecutional measures had to be faced by a Jewish person who was neither imprisoned nor had gone underground, but seemed to lead a normal life. Klemperer's records reveal an extreme kind of distress with permanent fear of death very close to that described by psychiatrists as victims of the Nazi persecution. It is then discussed how Klemperer experienced himself under these circumstances. Severe depression, distorted perception of time, stupefaction, egoism, and retreat from his compassion for fellow-sufferers may be understood from a psychodynamic point of view but were of course perceived as demoralization by Klemperer himself. Finally the present paper pursues the potentials of coming through and coping with persecution. Klemperer describes various ways and means of compensating and coping which were subsequently taken away from him with growing persecution. His non-Jewish wife was his only personal supporter. Writing was a constantly effective coping measure to him, initially as an essayist. After he had been banned from the academic community his diary remained his only opportunity for writing. In spite of being under constant fear for his life, he kept on writing. The subjective experiences of this situation are discussed. His recording of everyday life under persecution enabled him to survive it. Hence his diaries had a life-preserving function. PMID- 10478300 TI - [Drug therapy of hyperkinetic diseases in adults]. AB - Attention deficit hyperactivity disorder (ADHD) is often diagnosed in children. Some major studies proved that symptoms can persist until the patients are grown up and that the prevalence of adult ADHD is underestimated. The classification systems ICD-10 and DSM-IV allow a diagnosis for adults, they differ, however, from childhood regarding symptoms. The diagnostic procedure requires extensive exploration and information from parents and school. Diagnostic interview procedures and questionnaires for self and expert rating are available to date only for English speaking countries. Apart from counselling and psychotherapeutic intervention pharmacotherapy is a third option. Especially stimulants proved to be an efficient treatment. Alternatively tricyclic antidepressants can be considered, other agents play only a minor role. Clinical trials trying to evaluate the efficacy of different pharmacotherapeutic agents are lacking. PMID- 10478301 TI - [Karl Ludwig Kahlbaum--his life and work until his recognition as a psychiatrist. The occasion of the 100th anniversary of his death, April 15, 1999]. AB - Papers by Harms [3] and Thieme [19] acknowledge Kahlbaum as a pioneer in the field of children and adolescent psychiatry as well as the work in his asylum in Gorlitz, whereas Katzenstein [8] analyses his contribution to the development of psychiatry as theory. The present article is meant as a supplement to these. It is devoted to Kahlbaum's life before he became accepted as a therapist as well as scientist. As a result, first of all by having investigated his student time in Leipzig, it can prove conjectures that Kahlbaum became interested in psychiatric issues rather late. Furthermore the paper reconstructs the discussion lead around his most well-known scientific work, his nosology of 1863, and substantiates the fact that its author experienced appreciation in this particular field only decades later. PMID- 10478302 TI - [Sports injuries of the nervous system]. AB - Almost 1% of all Germans suffer sports injuries each year, almost 5% of all peripheral nerve lesions are due to sports. A review is given on various activities detailing the specific risks for traumata of the central and peripheral nervous system. Specifically these are volleyball, handball, basketball, American football, soccer, bowling, hockey, baseball, tennis, golf, javelin, fencing, wrestling, judo, boxing, running, jumping, dancing, mountain climbing, weight lifting, gymnastics, horse-back riding, swimming, rowing, skiing, skating, shooting, (motor) biking, car racing, flying, and sports for the disabled. The knowledge of typical traumata should enable the neurologist to rapidly and reliably recognize related lesions and to contribute to their prevention or improvement. PMID- 10478303 TI - [M Adli, T.Bschor, B. Canata. S. Dopfmer, M. Bauer: Lithium in the treatment of acute depression. Fortschr Neurol Psychiat 199866: 43]. PMID- 10478304 TI - Problems of female sexuality: the defensive function of certain phantasies about the body. AB - The author discusses certain modalities employed by the female mind when entering into a relationship with the representations of the self and with those of her own anatomy, i.e. with representations of internal and external reality. The hypothesis is that for women in whom representations of a fragile self predominate, the body is not only felt as a mysterious, receptive cavity, but also as an unreliable organism. Here an equation between the precariousness of the self and that of the body seems to be established and the female body is felt as a wounded body, which, through an opening over which there is no control, may lose its inner contents and let in dangerous aggressors. The absence of a penis, which is discovered later, may become the symbolic representation of this precariousness. The vaginal opening may also be seen in a way that triggers persecutory anxieties. The phantasy that defends against these painful perceptions may be that of secretly and guilty having some qualities of a male body, which are represented by reassuring male sexual attributes. In borderline and psychotic syndromes, the phantasy of having a male body appears to become concrete and almost delusional. In one case of female homosexuality, the unconsciously active phantasy is that of having the attributes of both sexes. These phantasies are defensive manoeuvres connected with early self-loss and non integration anxieties. They are far removed from the maternal and paternal identifications that should pave the way to mature, psychic bisexuality; that is, to the presence of male and female psychic attributes and to the availability of the cathected object of both sexes. PMID- 10478305 TI - Absolute zero: a man who doubts his own love.... AB - In this paper the author explores the relationship, first formulated by Freud, between depression and the experience of losing one's feeling of being able to love. She emphasises the narcissistic organisation underlying depression and depressive anxieties, and argues that the defences used to protect against such anxieties frequently produce the very states of mind they are supposed to be defending against. Using clinical material from a patient in analysis, she describes the way in which a combination of idealisation and a subtle, pervasive contempt for objects creates a brittle, sham relationship with the analyst, and that when this sham relationship is threatened, the patient can feel a catastrophic sense of desolation. Such events threaten the analysis, and often create correspondingly great anxiety in the analyst. At such times the analyst may be tempted to return the analytic relationship to the earlier, seemingly less disturbing state. The author argues that this would be a mistake, because it perpetuates the patient's despair about being able to engage only in fraudulent, ultimately empty relationships, and his fear that the analyst is unable to recognise this despair and to help him with it. PMID- 10478306 TI - Cynicism: its function in the perversions. AB - The author begins by noting the historical importance of perversion in the development of psychoanalysis and its potential for inspiring new ideas. Observing that cynicism is a component of perversion, he discusses some psychoanalytic views of the cynic and the role of the cynical attitude in politics. After a brief reference to the philosophy of the ancient Cynics and to irony, which he distinguishes from cynicism proper, he considers the clinical and theoretical aspects of cynicism and perversion in terms of representation/affect and hallucination/sensual pleasure. The connections with fetishim in particular are explored. The cynic is stated to possess an ethic of the negative, in which beauty is trampled underfoot by linguistic acts permeated with the subject's internal void, and to use a sexual theory translated into ideology. Specific elements of the cynic's logic include, in the author's view, a vindication of the mother's phallus, scepticism about the possibility of a male identification in the patient's father, and premonitions of the inexorable return of the non-human. These points, as well as the radical differences between perversion and cynicism on the one hand and neurosis and psychosis on the other, are illustrated by a literary example and the case history of a frotteur. PMID- 10478307 TI - Some notes on the 'heroic self' and the meaning and importance of its reparation for the creative process and the creative personality. AB - The author discusses a few points in a long clinical history from which he has derived the notion of 'heroic self', describing how it manifested through 'heroic' projective and introjective identifications. He also demonstrates the importance of repairing both the heroic self and the internal objects, and how essential it is for the analyst to differentiate between the heroic self and its megalomaniac psychopathic manifestations, and to recover the heroic self when it appears to be lost, as in the case of severe, quasi-melancholic, depression. By heroic self and heroic projective and introjective identifications, he means the creative person's specific need to associate himself with, to compete with and excel the heroes of his own cultural tradition. The creative person is able to do so via these projections and introjections, and the need is based on the creator's inner awareness of naturally possessing unusual, or even outstanding, gifts; in other words, the creative person has an awareness of being, or of the possibility of becoming, a creative hero. Brief reference is made throughout to other clinical material, and in the final section further support is given to the author's views on the heroic self with short quotations and examples from non psychoanalytic sources that provide further interesting evidence of the existence and functioning of the heroic self. With particular reference to Segal's work in this field (1952, 1991), and her notion of 'meaningful form', the author tries to further develop its implications in the light of the heroic self and of the heroic projective and introjective identifications, as far as the creator and those who respond to and evaluate his or her work are concerned. PMID- 10478308 TI - Listening, understanding and interpreting: reflections on complexity. AB - Beginning with the premise that a contemporary approach to analytic exchanges has become far more complex and multi-faceted than in earlier times, the author addresses listening, understanding and interpreting. The opening section presents the basis and utility of a proposed theory of five motivational systems and identifies the need for a shift from conceptualising 'structures' to systems. The nature of communication during analysis is considered from several standpoints including listening for needs and intentions, the place of theory as a background to listening, an optimal state for analysand and analyst, and the significance of the distinction between inner monologue and spoken discourse. Differing views of free association and narrative, especially questions arising from findings of the 'adult attachment interview', are discussed. The patient's sensitivity to the presence and influence of the analyst and the analyst's recognition of non-verbal as well as verbal communications completes this section. In the final section a brief clinical example is presented to introduce a differentiated depiction of the variety of interventions that analysts employ. Throughout the paper, the author presents his view of listening, understanding and interpreting in dialectic contrast with the many other perspectives held by analysts in this period of theoretical pluralism. PMID- 10478309 TI - The psychoanalytic psychotherapy of a mentally retarded man. AB - In this paper, the author discusses the psychoanalytically informed treatment of a 25-year-old patient, who at the beginning of treatment was severely developmentally retarded. The patient showed repetitive behaviours (tapping his fingers, tying and retying his shoelaces); visual and auditory hallucinations, delusions, vague to absent ego boundaries; inability to do simple maths, tell the time, read or write; and angry vulgar outbursts accompanied by occasional throwing of objects. He was bewildering and unresponsive to various psychological and physical treatments that had been performed since his infancy. The patient had been examined by a variety of medical and psychological specialists, but he seemed to belong in no particular place in the system of health-care providers. All courses of drugs that were tried proved ineffective. This author presents clinical material from a psychoanalytic psychotherapy that resulted in remarkable improvement in the patient's capacity for self-reflective thought as well as in his capacities for more mature object relationships and work. More specifically, the author discusses aspects of the treatment he thinks were most pivotal: (1) the interpretation of sensation (autistic) states; (2) the use of countertransference derived understandings of the patient's intra- and intersubjective experiences to guide his interventions; (3) the use of sensation based communication in a verbal form of play. PMID- 10478310 TI - 'My heart belongs to daddy'. Some reflections on the difference between generations as the organiser of the triangular structure of the mind. AB - The author begins his paper with a historical review of the concept of the difference between generations, which is in his opinion a metaphorical transformation that underpins the three-dimensional functioning of the psychic apparatus by introducing a differentiating intergenerational space between subject and object. He postulates that at the point of intersection between the intersubjective and the intrapsychic the subject clings to the specific fragments of his parents' history that are consistent with a belief about himself and the oedipal couple in which intergenerational links are severed and infantile incestuous wishes are seen as fulfilled. Disavowal of this generation gap is considered to lead to failure of post-oedipal secondary identifications, resulting in disturbance of the triangular structuring of the mind and consequent impairment of the genesis of thought processes. These ideas are compared with related conceptions of other authors and illustrated, with an account of the associated transference/countertransference vicissitudes, by a clinical example of the constellation the author calls 'My heart belongs to daddy', which he sees as a way station in the negotiation of the female Oedipus complex. PMID- 10478311 TI - Freud's early clinical theory (1894-1896): outline and context. AB - The author points out that all Freud's publications and lectures from the period 1894-1896 had the aim of establishing an aetiological theory of the neuroses and psychoses. Freud found that the clinical pictures covered by the theory had a sexual aetiology and were distinguishable from each other by specific mechanisms. Numerous references are presented to show that what the author calls Freud's early clinical theory had its roots in contemporary neuropathology and psychiatry, and the prevailing views in these disciplines are discussed. He was able to build on the work of predecessors such as Mobius in regard to the causal role of psychic factors. Whereas Freud was of his time in attempting to redefine the category of 'neurasthenia' and adducing the familiar concept of 'sexual noxae', his originality lay in the elevation of these factors, hitherto seen as external, to the status of 'true' causes. In the author's view, the rejection met with by Freud's early clinical theory was due to its pre-experiential 'vision' of the sexual aetiology of the neuroses rather than to its content. She notes that Freud differed from his contemporaries on the concept of and systematic part played by sexuality, as well as on the central role of psychic mechanisms (i.e. of defence). PMID- 10478312 TI - The Piggle: confrontations with non-existence in childhood. AB - The author interweaves pieces from D. W. Winnicott's 'The Piggle: An Account of the Psychoanalytic Treatment of a Little Girl' with her own experiences and case material, to explore how confrontations with existence/non-existence can become terrifying realities that overwhelm the child's resources. In the pages of 'The Piggle', we can see a young child 'playing' with the unfathomable notions of beginnings and endings in the wake of the birth of a sibling, which also heralds the death of the extant mother/child relationship. In the case material these issues are explored from the perspective of an adult who had been unable to resolve them without great loss of self. In both cases, parental failures impeded the child's ability to integrate his or her experiences. The work of Matte-Blanco provides a useful terminology for understanding condensations and equivalences in the material; as affect intensifies, the rules of conventional logic recede, and sameness threatens to annihilate distinctions between good and bad, or self and other. Containing these dichotomies within the analytic setting facilitates their integration, in a movement from the fragmentation of the paranoid-schizoid position towards the reconciliation and renunciation of the depressive position. PMID- 10478313 TI - Countertransference. PMID- 10478314 TI - 'What is "applied" in "applied" psychoanalysis?'. PMID- 10478315 TI - '"Dreams that turn over a page": integration dreams with paradoxical regressive content' by Jean-Michel Quinodoz. PMID- 10478316 TI - Role of routine diagnostic laparoscopy in the investigation of infertility. PMID- 10478317 TI - Reproductive outcome after sterilization reversal in women of advanced reproductive age. AB - OBJECTIVE: Our objective was to assess the clinical outcome of tubal reversal in women of advanced reproductive age. METHODS: A multicenter retrospective chart review of 153 patients who underwent a tubal ligation reversal was carried out. Patients were evaluated according to age. All patients had documented ovulation and a partner with a normal semen analysis by WHO criteria. Outcome measures included rates of clinical pregnancy, ectopic pregnancy, spontaneous abortion, and live birth, and the time to conception. RESULTS: Clinical pregnancy rates were significantly lower in women > or = 40 compared to younger groups. The time to conception was significantly shorter for women < 30 compared to women > or = 35. No pregnancies occurred in women > or = 42. CONCLUSIONS: Our data support the judicious use of sterilization reversal for infertile women with no male factor through their early forties. Women > or = 42 years should be especially counseled as to the very low success rates. PMID- 10478318 TI - The outcome of cryopreserved human embryos after intracytoplasmic sperm injection and traditional IVF. AB - OBJECTIVE: Our objective was to analyze the outcome of cryopreserved embryos obtained after intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) in terms of survival rate, implantation rate (IR), total and clinical pregnancy rate (PR) in a retrospective, comparative study. METHODS: Three hundred seventy-five IVF and 463 ICSI surnumerary cleaved embryos, frozen on Day 2 with 1,2-propanediol, were thawed. RESULTS: Thirty-two percent of the thawed IVF embryos survived and 11 pregnancies (8 clinical) were obtained from 68 transfers (16.1%). Fourty-seven percent of the ICSI embryos survived, with 19 pregnancies (18 clinical) from 116 transfers (16.4%). The IR was 8.5% (8/94) in IVF cycles and 10.8% (20/185) in ICSI cycles. CONCLUSIONS: A significantly better survival rate of ICSI embryos was observed but with no difference in PR, preclinical, and clinical abortion rate, or IR. PMID- 10478319 TI - The effect of endometrial polyps on outcomes of in vitro fertilization (IVF) cycles. AB - PURPOSE: Our purpose was to investigate the effect of endometrial polyps on pregnancy outcome in an in vitro fertilization (IVF) program. METHODS: Endometrial polyps less than 2 cm in diameter were suspected by transvaginal ultrasound before oocyte recovery in 83 patients. Forty-nine women (Group I) had standard IVF-embryo transfer, while in 34 women (Group II) hysteroscopy and polypectomy were performed immediately following oocyte retrieval, the suitable embryos were all frozen, and the replacement cycle took place a few months later. RESULTS: Of the 32 hysteroscopies, a polyp was diagnosed in 24 cases (75%) and polypoid endometrium in another 5 patients (15.6%). An endometrial polyp was confirmed by histopathological examination in 14 women (58.3%). The pregnancy rate in group I was similar to the general pregnancy rate of our unit over the same period (22.4 vs 23.4%) but the miscarriage rate was higher (27.3 vs 10.7%, P = 0.08). In Group II, the pregnancy and miscarriage rates were similar to those of the frozen embryo cycles at Bourn Hall (30.4 and 14.3 vs 22.3 and 12.1%, respectively). CONCLUSIONS: Small endometrial polyps, less than 2 cm, do not decrease the pregnancy rate, but there is a trend toward increased pregnancy loss. A policy of oocyte retrieval, polypectomy, freezing the embryos, and replacing them in the future might increase the "take-home baby" rate. PMID- 10478320 TI - Controlled ovarian hyperstimulation adversely affects implantation following in vitro fertilization-embryo transfer. AB - PURPOSE: Our purpose was to determine if controlled ovarian hyperstimulation adversely affects implantation. METHODS: A retrospective comparison of pregnancy rates (PRs) and implantation rates was made between oocyte recipients versus their donors, who shared half of the retrieved oocytes, and regular patients undergoing in vitro fertilization-embryo transfer (IVF-ET) who were not sharing eggs. RESULTS: Higher implantation rates (39.0 vs 22.5%; P < 0.05) were found in recipients compared to donors in the stimulated cycle. However, no differences were seen in PRs or implantation rates in frozen ET cycles. The data for standard IVF patients were almost-identical to those for donors. CONCLUSIONS: Superior implantation rates and PRs in oocyte recipients versus donors were not related to better oocyte quality for recipients because of egg sharing or to a better uterine environment because of similar results with frozen ET in all three groups. An adverse effect of the hyperstimulation regimen best explains the difference. PMID- 10478321 TI - Effects of hydrosalpingeal fluid on murine embryo development and implantation. AB - PURPOSE: Our purpose was to evaluate the effects of various concentrations of hydrosalpingeal fluid (HSF) on the preimplantation development and implantation of murine embryos. METHODS: One-cell mouse embryos were cultured in KSOM culture medium with 0.1, 1.0, 10, or 50% HSF, without and with lactate supplementation. Late-stage embryos were transferred into the uteri of pseudopregnant CD-1 females to determine implantation rates. The embryo transfer technique used was developed by our group and its effectiveness was evaluated during this experiment. RESULTS: Blastocyst development in the 0.1, 1.0, 10, and 50% group was 45, 55.0, 12.5, and 17.5%, respectively, with lactate supplementation, and 35.0, 52.5, 12.5, and 5.0%, respectively, without lactate supplementation, while in the KSOM (control) group it was 63.8%. Blastocyst development was reduced compared to controls in the 10% HSF and 50% HSF groups. Implantation rates for the 0.1 and 1.0% groups with lactate supplementation were 43.0 and 25.0%, respectively, and those with lactate supplementation were 50.6 and 61.8%, respectively, while in the KSOM group the implantation rate was 65.5%. None of the implantation rates were significantly different. CONCLUSIONS: Hydrosalpingeal fluid has a concentration dependent inhibitory effect on in vitro murine embryo development, but it has minimal effects on implantation rates. PMID- 10478322 TI - Requirements for human chorionic gonadotropin and recombinant human luteinizing hormone for follicular development and maturation. AB - PURPOSE: Our purpose was to evaluate the requirements for human chorionic gonadotropin (hCG) and recombinant luteinizing hormone (rec-LH) for follicular development and maturation in mice. METHODS: We carried out ovarian stimulation of immature mice. Output parameters were the preembryos created in vivo and frequency of blastocyst formation in vitro. RESULTS: hCG at 0 to 1 IU resulted in a dose-dependent recovery of preembryos (0 to 39.7 +/- 4.3; mean +/- SE) per mouse. hCG at 1 and 10 hCG gave similar results, whereas higher doses significantly reduced the number of preembryos. Potential for blastocyst formation was independent of hCG dose. hCG and rec-LH together exerted a synergistic effect on the recovery of preembryos. CONCLUSIONS: Optimal follicular development required a combination of 20 IU follicle stimulating hormone and 1-10 IU hCG. The potency of hCG was higher than that of rec-LH, but a synergistic effect of rec-LH and hCG was observed. The results may be pertinent for the development of strategies for ovarian stimulation of women with low levels of endogenous LH. PMID- 10478323 TI - The outcome of sperm retrieval and intracytoplasmic sperm injection in patients with obstructive azoospermia: impact of previous tuberculous epididymitis. AB - PURPOSE: Our purpose was to investigate the influence of previous tuberculous epididymitis in patients with obstructive azoospermia on the outcome of sperm retrieval and intracytoplasmic sperm injection (ICSI). METHODS: Eighty-eight cycles of ICSI were performed in 44 patients with obstructive azoospermia; 16 cycles (7 patients) with tuberculous obstructive azoospermia and 72 cycles (37 patients) with nontuberculous obstructive azoospermia. RESULTS: The rates of fertilization and embryo cleavage were comparable, and there was no significant difference in the clinical pregnancy rate per fresh transfer between the two groups. The rates of embryo implantation and clinical miscarriage were also comparable. CONCLUSIONS: Embryo quality and pregnancy outcome in sperm retrieval and ICSI were comparable in both the tuberculous and the nontuberculous obstructive azoospermia patients. Although there was a preponderance of testicular sperm used in the tuberculous obstructive azoospermia group, our results suggest that previous tuberculous epididymitis in patients with obstructive azoospermia does not affect the outcome of sperm retrieval and ICSI. PMID- 10478324 TI - Assessment of the dominant abnormal form is useful for predicting the outcome of intracytoplasmic sperm injection in the case of severe teratozoospermia. AB - PURPOSE: Our purpose was to investigate the relation between the dominant sperm anomaly and sperm morphology and the outcome of intracytoplasmic sperm injection (ICSI). METHODS: Two hundred ninety-five patients who underwent a total of 181 cycles of in vitro fertilization (n = 168) and/or 217 cycles of ICSI (n = 177) between July 1995 and May 1997 at Keio University Hospital were investigated. RESULTS: The rates of fertilization and pregnancy were 63.3 and 27.8%, respectively, in ICSI cycles with < or = 4% normal forms. When the percentage of strictly normal morphology was < or = 4, the fertilization rate was lower in the case of severely tapered head (13.0%; n = 4) than in the cases of other deformities in ICSI. The acrosomal defect made no difference in the fertilization rate with ICSI. CONCLUSIONS: The predominant abnormal form affects the ICSI outcome in the case of < or = 4% normal forms. PMID- 10478325 TI - Sperm artificially exposed to antisperm antibodies show altered deoxyribonucleic acid. AB - PURPOSE: Our purpose was to assess sperm deoxyribonucleic acid (DNA) integrity after exposure to antisperm antibodies. METHODS: Donor semen were divided and exposed to sera containing IgG, IgA, and IgM antisperm antibodies. Untreated portions served as the control. After incubation (1 hr, 23 degrees C), the sperm were centrifuge-washed, resuspended, and incubated (23 degrees C) for 2, 5, 7, or 9 days. Acridine orange staining and kinematic parameters were measured. The sentinel (17q21 from D17S855) and beta-globin genes were amplified and analyzed using denaturing gradient gel electrophoresis. RESULTS: Sperm preexposed to antisperm antibodies had deleted sentinel gene on days 7 and 9. The beta-globin gene was intact. There were no differences in acridine orange staining. CONCLUSIONS: Sperm artificially exposed to antisperm antibodies resulted in a subtle deletion of genetic material. The DNA alteration process was slow and was undetectable at the gross level. More studies are needed to confirm the findings and determine whether DNA repair mechanisms can reverse the damage. PMID- 10478326 TI - Periovulatory serum inhibin-a in women with a diminished ovarian reserve. PMID- 10478327 TI - Response to Hovav et al. [Diagnostic laparoscopy in primary and secondary infertility (J Assist Reprod Genet 1998;15:535-537)]. PMID- 10478328 TI - Effect of ascorbic acid supplementation on liver and kidney toxicity in cyclophosphamide-treated female albino rats. AB - Effects of ascorbic acid supplementation on the activity of acid phosphatase (ACP), alkaline phosphatase (ALP), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) on liver, kidney and serum in cyclophosphamide-treated female virgin rats were investigated. Oral administration of cyclophosphamide at the dose of 5 mg/kg body weight/day for 12 days resulted in a significant elevation in ACP and ALP activities in liver, kidney and serum. Ascorbic acid supplementation at the dose of 25 mg/kg body weight/day showed a significant protection in the activity of ACP in liver, kidney and serum, but only in ALP activity in kidney. ALP activities in liver and serum were not restored to control level by ascorbic acid supplementation. Activities of GOT and GPT were elevated significantly in liver, kidney and serum after cyclophosphamide treatment, and were protected and restored to control level by ascorbic acid supplementation. PMID- 10478329 TI - Time course observation of thyroid proliferative lesions and serum levels of related hormones in rats treated with kojic acid after DHPN initiation. AB - Time course changes in thyroid proliferative lesions as well as related hormone levels in the blood of male F344 rats given N-bis(2-hydroxypropyl)nitrosamine (DHPN: 2800 mg/kg body weight, single s.c. injection) as an initiation treatment followed by pulverized basal diet containing 0% (Group 2), 2% (Group 3) or 4% (Group 4) kojic acid (KA) were examined at Weeks 1, 2, 4, 8 and 12. As an untreated control group (Group 1), rats were given basal diet for 13 weeks and examined in the same manner. Serum T3/T4 levels in the DHPN + 2% KA and DHPN + 4% KA groups were significantly reduced as compared with the DHPN-alone group at each time point. Serum TSH levels in both DHPN + KA groups were significantly increased at each time point in a treatment period-dependent manner from Weeks 1 to 12, and the extent of elevation was more remarkable in the DHPN + 4% KA group. At Week 2, there were no statistically significant intergroup differences in liver T4-UDP-GT activities on a milligram microsomal protein basis. Histopathologically, no thyroid proliferative lesions were observed in the untreated control group or the DHPN-alone group. However, diffuse follicular cell hypertrophy and decreased colloid in the thyroid were apparent in all rats of the DHPN + KA groups at each time point. In addition, focal follicular cell hyperplasias and adenomas of the thyroid were observed at high incidence in the DHPN + 2% KA group from Week 4 and in the DHPN + 4% KA group from Week 8. Multiplicities of focal follicular cell hyperplasias and adenomas of the thyroid in the DHPN + 2% KA group were significantly greater than those in the DHPN + 4% KA group at Week 8. In the pituitary, an increase in the number of TSH producing cells with expanded cytoplasm was apparent from Weeks 4 to 12 in both DHPN + KA groups. These results strongly suggest that thyroid proliferative lesions were induced by KA administration due to continuous serum TSH stimulation through the negative feedback mechanism of the pituitary-thyroid axis, resulting from depression of serum T3 and T4. PMID- 10478330 TI - Effect of carbaryl on some biochemical constituents of the blood and liver of Clarias batrachus, a fresh-water teleost. AB - Carbaryl, a carbamate pesticide, (LC50 15.08 mg/l for 96 hr, i.e. lethal concentration with 50% mortality) induced perturbations in the levels of certain biochemical components including the activities of some enzymes in the blood and liver of the fresh-water catfish, Clarias batrachus exposed to sublethal concentrations (1, 2 and 4 mg/l) of the pesticide for 96 hr and 15 days. The pesticide caused a decrease in the levels of total protein and glucose with a concomitant increase in the levels of inorganic phosphate and lactic acid in fish serum. However, very little change was recorded in the serum cholesterol level. The treatment of the fish with carbaryl led to a marked increase in the activities of transaminases (GOT and GPT), phosphatases (acid and alkaline) and lactate dehydrogenase in the fish serum, the magnitude of the effect being dependent on the pesticide concentration and duration of exposure. The increase in lactic acid concentration with subsequent decrease in glucose concentration indicates an enhanced rate of glycolysis due to pesticide stress. Furthermore, the significant decrease in the activity of fish liver succinate dehydrogenase suggests that anaerobic metabolism was favored over aerobic oxidation of glucose through Kreb's cycle in order to mitigate the energy crisis for survival. The rise in the activities of transaminases and acid phosphatase due to pesticide intoxication suggest enhanced protein catabolism and probable hepatocellular damage in the organism. PMID- 10478331 TI - Toxicological response of rats to a novel monoamine oxidase type-A inhibitor, (5R)-3-[2-((1S)-3-cyano-1-hydroxypropyl)benzothiazol-6-yl]-5- methoxymethyl-2 oxazolidinone (E2011), orally administered for 13 weeks. AB - (5R)-3-[2-((1S)-3-cyano-1-hydroxypropyl)benzothiazol-6-yl]-5- methoxymethyl-2 oxazolidinone (E2011) is a novel monoamine oxidase type-A (MAO-A) inhibitor. In order to assess toxicological profiles of E2011, doses of 0 (as controls), 30, 100 mg/kg of E2011 were administered to male and female Sprague-Dawley rats once a day for 13 weeks orally by gavage. No mortality or any toxic signs except salivation occurred due to E2011 treatment. Decreased body weight gain and food consumption, increases of alkaline phosphatase and increases of liver weight were the major treatment-related findings observed predominantly in the 100 mg/kg group. Histological examination revealed nuclear enlargement of hepatocytes with appearance of altered cell foci in some cases, and acinar atrophy in Harderian glands in the 100 mg/kg group. Since the histopathological findings in the liver were indicative of an ongoing carcinogenic process, glutathione S-transferase placental form (GST-P) positive hepatic foci were identified immunohistochemically and examined morphometrically. Although GST-P positive hepatic foci were detected in all groups including controls, the number and area of GST-P positive hepatic foci were significantly higher in female rats treated with 100 mg/kg than those in controls. In this paper, possible mechanisms of specific lesions in the liver and Harderian glands will be discussed. PMID- 10478332 TI - Dose-dependent induction of carcinomas and glutathione S-transferase placental form negative eosinophilic foci in the rat liver by di(2-ethylhexyl)phthalate after diethylnitrosamine initiation. AB - The dose-dependence of di(2-ethylhexyl)phthalate (DEHP) hepatocarcinogenicity was investigated in male F344 rats which were initially injected with diethylnitrosamine (200 mg/kg, i.p.) and subjected to partial hepatectomy at week 3. The animals were administered DEHP in the diet at concentrations of 30, 300, 3,000, or 12,000 ppm starting 2 weeks after the DEN injection for up to 46 weeks and killed at weeks 8, 24, 48 and 52. Additional groups were given clofibrate (3,000 ppm in diet) or basal diet instead of the DEHP diet. Incidences of hepatocellular carcinomas were 75% (9/12, P < 0.01) for 12,000 ppm, 10% (1/10) for 3,000 ppm, 7% (1/14) for 300 ppm, 0% (0/13) for 30 ppm, 15% (2/13) for clofibrate, and 8% (1/13) for the basal diet group at week 52, 4 weeks after cessation of chemical feeding. Development of glutathione S-transferase placental form (GST-P) positive foci was only slightly increased by clofibrate administration at week 52 and consistently lower than the control level in the DEHP-treated groups after 24 weeks. In contrast, GST-P negative eosinophilic foci were dose-dependently increased in the more than 300 ppm DEHP and clofibrate treated groups. At the 30 ppm dose level, however, no morphological changes were apparent in the liver. Thus, the non-observed effect level regarding the promotional activity of hepatocarcinogenesis was demonstrated at 30 ppm, the effects being predictable on the basis of development of GST-P negative eosinophilic foci. PMID- 10478334 TI - Availability of sperm examination for male reproductive toxicities in rats treated with boric acid. AB - Sperm morphological examination, computer-assisted sperm analysis (CASA) and histopathologic examination of the testis and epididymis were performed for male rats treated orally with boric acid for 3 weeks at dosage levels of 50, 150 and 500 mg/kg/day, and treated males were mated with untreated females. None of the males treated with 500 mg/kg/day could impregnate untreated females. The fertility index showed a tendency to decrease in males treated with 150 mg/kg/day. At necropsy, the pre-implantation loss rate in females mated with males treated with 150 mg/kg/day was higher than the control values. Upon epididymal sperm analysis using the CASA system, all parameters including the number of sperm and sperm motions were found to be affected in males treated with 500 mg/kg/day, and the number of sperm, percent motile, velocities and amplitude of lateral head displacement (ALH) were affected in males treated with 150 mg/kg/day. Upon sperm morphological examination, head and tail abnormalities were observed in males treated with 150 and 500 mg/kg/day. In the histopathological examination, atrophy of the seminiferous tubules and multinucleated giant cells in the testes were observed in males treated with 500 mg/kg/day. PMID- 10478333 TI - Characterization of epididymal sperm motion and its correlation with stages of target cells in rats given alpha-chlorohydrin, cyclophosphamide or nitrazepam. AB - Epididymal sperm motion in rats was characterized by computer-aided sperm motion analysis (CASA) with its correlation to testicular lesions in the 2-week treatment study, using three compounds which are known to affect different stages of germ cells. Mature male rats were treated daily for 2 weeks with alpha chlorohydrin (alpha-CH, 5 mg/kg), cyclophosphamide (CP, 20 mg/kg) or nitrazepam (NZ, 20, 40, 60 mg/kg). Changes in sperm motion were detected only in the alpha CH and 60-mg/kg NZ-treated groups. Of the sperm motion parameters, velocity and amplitude of lateral head displacement (ALH) were concomitantly reduced in these two groups with good correlation. With respect to the distribution of the values in parameters, however, alpha-CH shifted the values down within a small range with high percentages of motile sperm, while NZ distributed them over a wide range with low percentages of motile sperm. CP treatment showed no histopathological changes in advanced germ cells, though it showed a decrease in the number of early germ cells. NZ treatment affected round and elongating spermatids (approximately step 14) at doses of 20 and 40 mg/kg, and affected also more advanced spermatids (approximately step 19) at the dose of 60 mg/kg. alpha CH treatment did not affect testicular histopathology. These findings indicate that 60-mg/kg NZ treatment reduced sperm motion as a result of lesions affected in elongated spermatids and alpha-CH reduced it by direct effects on epididymal spermatozoa. The present study indicates that in addition to percentage of motile sperm, the velocity and ALH can be useful to detect the changes in sperm motion caused by different actions of NZ and alpha-CH, though each compound showed a distinct distribution pattern of these parameters. PMID- 10478335 TI - A significant relationship between glutathione-peroxidase (GSH-PO) localization and biological action of testosterone in rat ventral prostate. AB - In order to confirm the relationship between glutathione-peroxidase (GSH-PO) localization and biological testosterone action in the rat ventral prostate, immunocytochemical localization of GSH-PO in glandular epithelial cells of the rat ventral prostate was investigated. In the untreated group, GSH-PO was predominantly demonstrated in glandular epithelial cells of the ventral prostate. Intracellular localization of GSH-PO in the glandular epithelial cells was mainly observed in cytoplasmic matrix near the rough endoplasmic reticulum and was occasionally noted as a small granular structure (GSH-PO-positive granule) at the supranuclear region. In a castrated animal, the intensity of GSH-PO staining in the glandular epithelial cells was remarkably decreased. By testosterone administration to the castrated animal, GSH-PO was clearly detected in the glandular epithelial cells. Intracellular localization of GSH-PO was mainly observed in cytoplasmic matrix and the number of GSH-PO-positive granules increased remarkably. These findings suggest that immunostainable GSH-PO in the glandular epithelial cells of the rat ventral prostate is testosterone-dependent, and that its staining pattern is a useful marker for biological testosterone action. PMID- 10478336 TI - Induction of unscheduled DNA synthesis in hairless mouse epidermis by skin carcinogens. AB - Induction of unscheduled DNA synthesis (UDS) in hairless mouse epidermis by six chemicals was determined in an in vivo-in vitro assay by using a liquid scintillation counting method. Test chemicals were applied once onto two areas of the back of female hairless mice after stripping of the stratum corneum with adhesive tape to enhance skin penetration. After exposure, the skin samples were taken and cultured in a medium containing [3H]thymidine with or without hydroxyurea (HU, an inhibitor of replicative DNA synthesis). DNA of the epidermis was extracted, and incorporation of [3H]thymidine into DNA and the DNA content was determined with a liquid scintillation counter and a fluorescence spectrophotometer, respectively. Induction of UDS by chemicals was judged by calculation of the UDS index [(the ratio of DNA synthesis in the presence of HU to that in its absence) x 100]. A good correlation between UDS induction and organ specificity of carcinogens was observed. 4-Nitroquinoline 1-oxide, a skin carcinogen used as a positive control, induced a dose-dependent increase in the UDS index of approximately 12-fold at 2 hr after exposure, while 1,2 epoxydodecane, a non-skin carcinogen applied as a negative control, did not increase the UDS index. Four other skin carcinogens induced dose-dependent increases in the UDS index; N-methyl-N'-nitro-N-nitrosoguanidine and diepoxybutane at 2 hr after exposure, and 7,12-dimethylbenz[a]anthracene and benzo[a]pyrene at 24 hr after exposure. The results suggest that UDS is a good marker of the genotoxicity of skin carcinogens. PMID- 10478337 TI - Role of cocaethylene in toxic symptoms due to repeated subcutaneous cocaine administration modified by oral doses of ethanol. AB - The present study investigated the toxicity of repeated subcutaneous cocaine administrations combined with oral doses of ethanol, and discussed the role of the toxic metabolite cocaethylene. Subcutaneous cocaine (70 mg/kg) was given to male ICR mice at 45 min after an oral administration of either ethanol (maximum 3 g/kg) (cocaine-ethanol group; n = 50) or saline control (cocaine group; n = 30), once per day, for up to 5 days. In the combined cocaine-ethanol group, the total frequency of death was significantly increased (86%) as compared to the cocaine group (40%). In both administration groups, regardless of the day of death, "late" deaths characterized by the late and unexpected onset of fatal symptoms could be differentiated from "early" deaths on the basis of the survival time after the last cocaine injection, the drug concentrations in the tissues at the time of death, and/or the observed physical disorders. In the combined cocaine ethanol group, a late death group with survival times exceeding 12 hr and two early death groups could be differentiated, based on the presence or absence of cocaethylene and the different types of clinical symptoms. In the early death group in which cocaethylene could be detected, the volume of ethanol ingested was not significantly different from the late death group with large ethanol consumption and slow exacerbation of the respiratory and locomotive symptoms. Furthermore, the severity of the cocaine-induced seizures was also similarly decreased by ethanol. In the other early death group in which cocaethylene could not be detected, the volume of ethanol ingested was significantly lower than in the late death group, and seizures as severe as in the cocaine-only group were observed. PMID- 10478339 TI - [Neural stem cells: application for the neural grafting]. PMID- 10478338 TI - Effects of di-isononyl phthalate (DINP) on peroxisomal markers in the marmoset DINP is not a peroxisome proliferator. AB - In the present study the systemic toxic potential of di-isononyl phthalate (DINP) was assessed in a 13-week study in marmosets. Particular attention was given to its potential for hepatic peroxisome proliferation. Three groups of four male and four female marmosets received DINP, by oral gavage administration, at dosages of 100, 500 or 2500 mg/kg/day for 13 weeks. A fourth group served as a concurrent Control group and received the vehicle (1% methylcellulose and 0.5% Tween) only. A fifth group received clofibrate at a dosage of 500 mg/kg/day to provide a positive Control for liver peroxisome activity. At the end of the treatment period, the animals were killed and their livers were removed. 3000 x g supernatant and microsomal subcellular fractions were prepared from homogenised liver by differential centrifugation. The peroxisomal marker enzyme activity, cyanide-insensitive palmitoyl CoA oxidase, was assayed in the former, while cytochrome P450 concentration and lauric acid 11- and 12-hydroxylase activities (selective for CYP2E1 and 4A, respectively) were assayed in the microsomes. No statistically significant changes were seen in any of these parameters measured following DINP treatment, compared with the Control. Clofibrate treatment resulted in an approximately 100% increase (p < 0.01) in both male and female marmoset cyanide-insensitive palmitoyl CoA oxidase activity and a similar increase (p < 0.05) in male (only) lauric acid 11-hydroxylase activity. No other changes were statistically significant at the 5% level. These data provide no evidence that DINP was acting as a peroxisome proliferator when administered to marmosets under the conditions of the study. PMID- 10478340 TI - [Techniques of facial nerve reanimation: unilateral hypoglossal-facial nerve anastomosis and great auricular nerve grafting]. PMID- 10478341 TI - [Microsurgical results of paraclinoid aneurysms of the internal carotid artery: microsurgery versus intravascular surgery]. AB - Since paraclinoid aneurysms exhibit considerable variations in the surrounding skull base structures, some skill and experience are considered essential for their microsurgical treatment. On the other hand, intravascular coil embolization would be free from these extravascular factors. Fifty-two aneurysms in 48 patients were treated microsurgically. They were divided into an early series treated before 1995 and a late series treated after 1996. Three aneurysms in 3 patients were treated by intravascular surgery. Preoperative neuroimages, topographic anatomy and surgical results were compared retrospectively. The results in the microsurgical late series were better than those in the early series. While 89% of the patients exhibited excellent results with two patients left with partial visual field defect (7%) and one death (3%) in the early series, the rates in the late series were 95%, 5% and 0%, respectively. Complications and failure in neck clipping were considered to be due to such topography as, 1) C3 aneurysms extending into the anterior clinoid process, 2) multiple aneurysms, 3) ophthalmic artery originating near the dome, 4) atheroma or calcification at the neck, 5) marked medical shift of C2 segment, and 6) tight adhesion of the dural ring to the dome. Coil embolization was successful in two aneurysms, while it was given up in one. We currently propose that intravascular surgery is indicated for cases 1), 4), 5) and 6), and microsurgery would be more advantageous for cases where the neck is broad and where the topography concerning aneurysmal multiplicity or the branching site of the ophthalmic artery is not fully understood by preoperative imaging. PMID- 10478343 TI - [Experimental study of water leakage in duraplasty using ePTFE sheet as an artificial dura mater]. AB - The authors have evaluated the preventive effect of water leakage from the suture holes of expanded polytetrafluoroethylene (ePTFE) sheet. Six currently available suture materials (4-0 SURGILON, 4 types of Gore-tex suture, DuraClose) were compared. The water leakage was 0.05 cc/min when using Gore-tex suture (CV-6, needle/thread ratio = 1.0), 0.025 cc/min when using DuraClose. With each two material, the water leakage was significantly less (p < 0.05) in comparison with other materials. Scanning electron microscopic examination revealed that the residual suture holes were minimum when using Gore-tex suture (CV-6, needle/thread ratio = 1.0). In conclusion, running suture with Gore-tex suture (CV-6, needle/thread ratio = 1.0) could be effective in preventing water leakage from the suture holes of ePTFE sheet in duraplasty. PMID- 10478342 TI - [Self-expanding stent (Wallstent) supported angioplasty for carotid artery stenosis]. AB - We reported the preliminary results of percutaneous transluminal angioplasty (PTA) supported with a self-expanding stent (Wallstent) for the cervical internal carotid artery (i.c.) stenosis in 6 patients (7 lesions). All were male aged from 60 to 79 (mean 70.8). Three patients were symptomatic and 3 were asymptomatic. The procedure was attempted in patients with severe i.c. stenosis according to criteria of NASCET, ECST or ACAS studies and with high risk if carotid endarterectomy (CEA) were performed. In all cases, vascular access was from the femoral artery and angioplasty was performed without cerebral protection. Successful angioplasty was obtained in all cases with no mortality. Stenosis was improved from 82.7% (74-90%) to 17.4% (10-33%). Perioperative complications were seen in only one case with TIA during the postdilatation period, which was supposed to be due to a distal embolism. The preliminary results of PTA supported with self-expanding stent for severe i.c. stenosis were excellent. These results suggested that some patients with severe i.c. stenosis can be treated safely and effectively by stent-supported PTA. However, it is necessary to mark the indications clear and strict, and to follow up these lesions after the procedure, because of the possibility of perioperative ischemic complications and restenosis. PMID- 10478344 TI - [Cigarette smoking increases the risk of developing a cerebral aneurysm and of subarachnoid hemorrhage]. AB - Despite recent advances in diagnostic and therapeutic techniques, subarachnoid hemorrhage (SAH) is still a serious condition associated with high mortality and morbidity. There are no effective treatments other than surgical intervention. However, another option for decreasing the occurrence of SAH may be prevention of aneurysms formation and of their rupture by controlling risk factors. Cigarette smoking has been recently shown to be one of the major risk factors for SAH. We investigated whether cigarette smoking increased the risk of developing cerebral aneurysms and of SAH. Degree of smoking was investigated in 182 patients with SAH and in 123 patients with an unruptured cerebral aneurysm incidentally detected during investigation of other diseases. Sixty-nine patients with other diseases who were shown to be free of cerebral aneurysms through MR angiography served as controls. Smoking significantly increased the risk of both aneurysm formation and SAH; The odds ratio for SAH was 2.4, and for unruptured cerebral aneurysm 1.7. Smoking especially increased the occurrence of SAH in women and in youngsters. However, smoking did not influence the occurrence of cerebral vasospasm and multiplicity of aneurysms. These data suggest the importance of avoiding smoking to prevent the occurrence of cerebral aneurysms and of SAH. PMID- 10478345 TI - [A case of an intracranial tuberculoma, mimicking meningioma, that developed during treatment with anti-tuberculous agents]. AB - A 54-year old female patient presented with intracranial tuberculoma attached to the dura mater during treatment with anti-tuberculous agents. Compared tomography (CT) and magnetic resonance imaging (MRI) revealed a lesion resembling a cystic meningioma. A granulomatous tumor that adhered to the cerebral cortex and dura was totally removed. Histological diagnosis was epithelioid granuloma. After removal of the tuberculoma and treatment with anti-tuberculous medications, the patient seemed to be cured, and regrowth of the tumor has not been observed to date. Tuberculoma mimicking meningioma is a very rare phenomenon. In the present study, we review intracranial tuberculomas and discuss their radiological features. PMID- 10478346 TI - [Brain surface clear cell ependymoma: case report]. AB - We report a case of brain surface clear cell ependymoma. A 13-year-old boy presented with complaints of right hypesthesia. Computed tomography and magnetic resonance image showed a left fronto-parietal cystic, calcified mass lesion. He underwent total resection of the tumor including cyst wall. The tumor located on the surface of the parietal lobe was sharply demarcated from the surrounding brain tissue and there was no continuity with the ventricular wall. Histological examination of the surgical specimens showed oligodendroglioma-like cells that had round unclei, clear cytoplasm which formed perivascular pseudorosettes, and immunoreactivity for glial fibrillary acidic protein (GFAP). Electromicroscopically, microvilli were seen. The findings were compatible with clear cell ependymoma. The cyst wall was lined with a layer of single cuboidal cells and, immunohistochemically, had no basal membrane. The inner surface of the cyst was positive for EMA, and the cuboidal cells were positive for GFAP. We discuss possible mechanisms for tumor growth in our case and the histogenesis of its cyst. PMID- 10478347 TI - [A case of sacral meningeal cyst with Marfan syndrome]. AB - A rare case of sacral meningeal cyst Marfan syndrome is reported. A 40-year-old male who had a past history of Marfan syndrome was admitted to Tokyo Metropolitan Ebara Hospital due to sensory disturbance in the right S1-2 nerve root area on October 1998. On admission, neurological deficits were sensory disturbance and decrease right Achilles reflex. Plain sacral X-ray was normal. Magnetic resonance imaging revealed a cystic mass in the intrasacral space. MR myelography showed the cyst followed the thecal sac. Signal intensity of the cystic mass was the same as the cerebrospinal fluid. In the prone position, laminectomy of the sacrum was carried out. The neck of the meningeal cyst was ligated with the help of an aneurysmal needle. The patient's sensory disturbance disappeared postoperatively. As MRI becomes more frequently used in Marfan syndrome, it is important to keep the presence of such an entity in mind. MR myelography made it easy to diagnose the meningeal cyst. PMID- 10478348 TI - [A case of hemorrhagic non traumatic arteriovenous fistula of the scalp]. AB - Arteriovenous malformation of the scalp in an uncommon disease and treatment for this lesion is difficult and controversial. A twenty-three-year-old male, who reported no history of head trauma, presented with spontaneous hemorrhage from the left temporal scalp. Neurologically he was normal except the tinnitus. Left external carotid angiography demonstrated arteriovenous malformation fed by the superior temporal and posterior auricular arteries. A dilated tortuous occipital artery was also recognized. The lesions were able to be surgically resected, alleviating intraoperative massive hemorrhage by proximal temporary occlusion of the external carotid artery at the left neck. Small feeding arteries from the meningeal artery which were not recognized on preoperative angiography were found at surgery. The arteriovenous fistula was successfully resected without any cosmetic problem. PMID- 10478350 TI - [Lissencephaly]. PMID- 10478349 TI - [Neural tube defects and holoprosencephaly]. PMID- 10478351 TI - [Polymicrogyria]. PMID- 10478352 TI - [Cortical dysplasia]. PMID- 10478353 TI - [Non-Alzheimer degenerative dementias]. PMID- 10478354 TI - [Effects of interelectrode separation in sensory nerve conduction studies]. AB - Sensory nerve action potentials (SNAPs) obtained by conventional sensory nerve conduction studies are greatly influenced by the distance between exploring and reference electrodes. Changes in SNAPs were investigated in 12 healthy median nerves using the orthodromic and antidromic sensory nerve conduction studies with 1-cm, 2-cm, 3-cm, 4-cm, and 5-cm of interelectrode distances. Peak-to-peak (orthodromic) or baseline-to-peak (antidromic) amplitudes and durations between the positive peaks were measured and each value was expressed as a ratio to the recording of 1-cm of interelectrode distance. SNAPs with maximum amplitude were obtained by more than 3-cm of interelectrode distance. Duration of SNAP was increased by an increase of interelectrode distance. Change in amplitude along with the interelectrode distance was larger in the orthodromic method than in the antidromic method. Contrariwise, change in duration was larger in the antidromic method. SNAPs with long duration had a gently sloping configuration, and each peak tended to be indistinct. So, 3-cm of active and reference interelectrode separation should be recommended in performing sensory nerve conduction studies between wrist and finger, because maximum amplitude and clear visualization of peaks are available. PMID- 10478355 TI - [Usefulness of T2* weighted magnetic resonance image in the diagnosis of head injury on chronic stage]. AB - T2* weighted gradient echo image has heightened diagnostic sensitivity to hemorrhage, which is attributed to magnetic susceptibility-induced static field inhomogeneities arising from paramagnetic blood breakdown products such as hemosiderin, which shorten T2*. We examined 4 cases of head injury in chronic stage by T2* weighted images and assessed the clinical application of this sequence for diagnosis of the intracranial lesion. All patients underwent CT scan on acute stage and long-term follow up was performed. In all cases, abnormally low signals in the brain and subarachnoid space were more conspicuous with the T2* weighted image than with any of the conventional sequences. In 2 cases with diffuse axonal injury, of which lesions were not detected on acute stage CT scan, hypointensity area was clearly demonstrated in the cerebral peduncle and corpus callosum on T2* weighted images. Selection of T2* weighted image into the routine MR examination of patients with chronic stage of the head injury is recommended. PMID- 10478356 TI - [Treatment of spontaneous intracranial hypotension with continuous epidural saline infusion: a case report]. AB - We describe that a 27-year-old woman with a severe postural headache for a month due to spontaneous intracranial hypotension was successfully treated with a continuous epidural saline infusion (CESI). She presented with sudden postural headache and back pain. On admission, CT scans of the head and thoracic spine were normal. Spontaneous intracranial hypotension (SIH) was suspected from her symptoms that her headache was worsen by the erect position and relieved by the supine position. She was treated with intravenous infusion (1000 ml/day), but her headache did not resolve. Three weeks later, MR images showed descent of the brain stem and cerebellar tonsil, effacement of the sulci and cisterns, subdural fluid collections and diffuse meningeal enhancement. A lumbar puncture was performed and yielded an opening CSF pressure of 75 mmH2O in the horizontal position. Radionucleotide cisternography showed early accumulation of nucleotide in the bladder, slow ascent along the spinal axis, less than expected activity over the cerebral convexities and early disappearance of radioactivity. As for the treatment, an epidural catheter was placed at the L 1-2 level and CESI of 15 ml/hour was started. Immediately the postural headache resolved and the next day she could walk. CESI was continued for 5 days. MR images after CESI showed normalization of the sulci and cisterns including spinal subarachnoid space, reelevation of the brain stem and cerebellar tonsil and improvement of meningeal enhancement. She remained headache free during 7 months of follow-up period. We propose this method as a safe and effective treatment for SIH. PMID- 10478357 TI - [A case of transient isoniazid-induced myoclonic seizures]. AB - We report a unique patient who had temporary isoniazid-induced myoclonic seizures. A 74-year-old man noticed involuntary movements in the upper extremities when he received isoniazid in pulmonary atypical mycobacteriosis late June 1996. Neurological examination revealed myoclonic jerking in the upper limbs. Although isoniazid treatment had been given to November 1996, the myoclonic seizures were spontaneously ameliorated at 2 weeks following the onset. The second administration of isoniazid was started for the exacerbation of pulmonary atypical mycobacteriosis in February 13, 1999. Immediately he developed myoclonic seizures in the upper extremities. Neurological examination showed myoclonic jerking in the upper extremities and cerebellar ataxia in the lower extremities. These neurological deficits were naturally improved within a week. Brain MRI suggested multiple lacunar infarction. EEG showed slow theta and sharp waves. Somatosensory evoked potential (SEP) revealed giant potentials. There was no renal or liver dysfunction. Serum vitamin B6 levels were decreased slightly. The metabolic studies of serum and cerebrospinal fluid isoniazid concentrations demonstrated slow inactivation of this agent. After the end of isoniazid treatment, EEG and SEP were normal. The results of these studies indicated that slow inactivator of isoniazid and its epileptogenic effects might contribute to the pathogenesis of myoclonic seizures in our patient. PMID- 10478358 TI - [A case of encephalomyeloneuritis and HTLV-I infection]. AB - The patient, a 65-year-old woman, had liver cirrhosis, and had blood transfusion at the age of 49 and 56. Early in September, 1989, she gradually developed numbness of the legs, staggering gait, and apathy with hallucination. In October, she became incontinent and unable to stand, and was admitted to Konan Hospital. On admission, she was disoriented with poor comprehension. Cranial nerves were intact except for horizontal nystagmus on lateral gaze. She had generalized areflexia without pathological reflex. Muscular forces were fairly preserved. Superficial sensations were diminished in the upper limbs as well as below Th-7 level. Deep sensation was abolished in the distal parts of the extremities with athetotic finger postures on arm rising. She had urinary and fecal incontinence. Results of routine laboratory examinations were non-contributory. Chest CT scan and sputum cytology were normal. CSF contained one cell/microliter, 95 mg/dl of protein with positive oligoclonal IgG bands. Anti-HTLV-I antibody was positive in serum and CSF. Urodynamic studies showed neurogenic bladder of supranuclear type. MNCV was slightly decreased. SNAP and SEP were not evoked. On sural nerve biopsy, the density of myelinated fibers was 720/mm2, and that of unmyelinated fibers, 26,978/mm2. ABR and VEP were abnormal. EEG showed diffuse theta waves with paroxysmal delta and sharp waves. T2-weighted MR images of the brain showed patchy areas of high signal intensity in the cerebral white matter. Soon after administration of methylprednisolone, her consciousness became clear. EEG normalized in 4 months. Twenty months after the onset, she became ambulant with crutch, but still has dysuria and sensory deficits in the hands and lower limbs. The possible relationship between encephalomyeloneuritis and HTLV-I infection was discussed. PMID- 10478359 TI - [Temporal lobe epilepsy associated with old intracerebral hemorrhage due to capillary telangiectasis in the temporal lobe: case report]. AB - A case of 45-year-old female, who presented with temporal lobe epilepsy was reported. The patient was found to have old intracerebral hemorrhage due to capillary telangiectasis in the temporal lobe. On the intraoperative electrocorticography, frequent paroxysmal activities were recorded independently both on the medial and lateral aspects of the temporal lobe. Even after resection of the lateral temporal lobe, frequent paroxysmal discharges were noted on the hippocampus. Histologically, there are astrogliosis and hemosiderin deposits in the white matter around the telangiectasis and the old hematoma. It is postulated that the hippocampus gained secondary epileptogenicity. PMID- 10478360 TI - [MRI findings of acute relapsing disseminated encephalomyelitis with late onset]. PMID- 10478361 TI - [Metastatic skull tumor]. PMID- 10478362 TI - [Manichaeism and pathology?]. PMID- 10478363 TI - [Grading of malignant soft tissue tumors. Historical development and current status]. AB - This article describes the historical development of the grading systems for malignant soft tissue tumors. The first attempts to grade these tumors were made in the middle of the nineteen century; a remarkable amount of activity in grading took place in the 1970s which reached a maximum in the 1980s. Reviewing the literature back to the first available publications, five phases in the development of the grading systems for malignant soft tissue tumors could be distinguished. Five commonly used systems were checked for comparing and handling of 339 patients with malignant soft tissue tumors. The use of two multifactorial systems, one simple and one complex, to grade malignant soft tissue tumors should be favored. PMID- 10478365 TI - [Chlamydia pneumoniae--a suspected pathogenetic factor in coronary arteriosclerosis]. PMID- 10478364 TI - [Chondromyxoid fibroma. Morphological variations, site, incidence, radiologic criteria and differential diagnosis]. AB - Chondromyxoid fibroma is a rare benign bone tumor, accounting for less than 1% of all bone tumors. The peak age incidence is the second and third decade of life. Chondromyxoid fibrom occurs in the metaphyseal parts of the major tubular bones, predominantly of the lower extremity. Roentgenograms show, in most cases, a well demarcated radiolucent lesion. The classic histological feature of a chondromyxoid fibroma is stellate or spindle-shaped cells arranged in lobules in a myxoid or chondroid background. Analysis of 40 chondromyxoid fibromas demonstrates the morphological variation of this tumor. Cases were examined for age distribution, localization, and radiological and histological features. In 85% we found the typical histomorphological pattern. Recurrence rate was 12.5%. In four cases the appearance was uncharacteristic and differentiation from other tumors such as chondroblastoma or chondrosarcoma was quite difficult. By adhering to strict histomorphological criteria, definite diagnosis of chondromyxoid fibroma can be made in most cases. PMID- 10478366 TI - [Highly differentiated squamous epithelial carcinoma as a late complication of post-traumatic osteomyelitis]. AB - Squamous cell carcinoma as a late complication of chronic osteomyelitis is a well known phenomenon in traumatology, often occurring as a consequence of bone fractures. The majority of cases are observed in men between 50 and 60 years of age. The time from onset of inflammatory bone disease to malignant transformation differs but usually takes 30 years. In general, prognosis is thought to be favorable when adequate surgical therapy is carried out. Nowadays, it is important to recall this condition, because it has become rare. The cases reported here illustrate the difficulties that may be encountered in diagnosing malignant transformation, especially in lesions that develop in deep tissue layers and which may be responsible for various biopsies failing to reveal the true pathology. PMID- 10478367 TI - [Comparative analysis of various standard immunohistochemical procedures]. AB - Immunohistochemical staining techniques, such as the APAAP, ABC, EnVision and Immuf1p4x methods, play a leading role in the search to solve the questions arising in clinical diagnosis. With the increase in number of techniques for diagnostic examination, the sarch for suitable method is becoming more and more important. The goal of this study was to compare these different treatments with regard to sensitivity and specificity. Furthermore, an analysis of time and cost was made. The primary antibodies used were CD20, CD30 and IgM, as well as receptor antibodies estrogen and progesterone. The variable costs of each technique were calculated based on selected examples. The APAAP method required the most time and was, at the same time, by far the most expensive treatment. The EnVision method required the least time and showed very good staining results, which were comparable to those of the ABC and ImmunoMax techniques. The staining costs could be reduced by about 95% and the staining intensity improved substantially by using expensive receptor antibodies in combination with the Immuno-Max method. This study illustrates the importance of knowing different examination techniques, as well as the costs arising by their usage. PMID- 10478368 TI - [German-Latvian cooperation in the pathology specialty]. AB - In the field of pathology there have been very close connections since the beginning of this century and a regular exchange of ideas between the pathologists of Latvia and Germany. Pathologists of Latvia were members of the German Association of Pathology. The stages of political changes in Europe very much influenced the exchanges of ideas. The following presentation will outline the activities and personalities in the sector of pathology in the time period until 1918--time of the Russian Czar Empire--, during the National State of Latvia and the Second World War (1918-1940), then during the Russian occupation of Latvia (1944-1991) and also the new time period after the political changes. Evaluated in this presentation the yearly support of the Baltic/German Symposiums, assisted by the IAP, which in addition to the activities in pathology also promote and encourage many personal contacts and friendships. PMID- 10478369 TI - [Direct cost of cerebrovascular disease during the first year of follow-up]. AB - INTRODUCTION AND OBJECTIVE: There are not sufficient studies analyzing the health costs of ictus in Spain. We carried out a prospective study to evaluate the health costs incurred by a person with a stroke during the first postictal year. PATIENTS AND METHODS: We included 118 patients from the Stroke Unit of the Hospital Universitario San Carlos in Madrid, admitted between 1 July and 31 December 1996. We studied 90 survivors, of an average age of 68 years, one year after having an stroke and specifically calculated the cost of an average period in hospital, neuroimaging tests, rehabilitation treatment, medical follow-up in the Outpatient Clinic, transport costs and the cost of medicines. RESULTS: The average cost patient/year was: hospital admission (418,203 ptas.), health transport (108,209 ptas.), cost of medicines (74,647 ptas.), follow-up visits (64,496 ptas.), neuroimaging (61,203 ptas.), rehabilitation (58,643 ptas.). The total cost was 79,930,719 ptas. and the average cost patient/year 888,119 ptas. during the first year following the ictus. The use of health resources depended on the variables: handicap (increased in patients with a score < 60 on the Barthel scale), average neurological deficit on the Scandinavian neurological scale and sex (cost greater in women). The clinical follow-up of total infarcts of the territory of the anterior circulation cost twice as much as follow-up in cases of lacunar infarcts. CONCLUSIONS: Cerebrovascular disease is expensive in terms of health-care. Fifty four percent of the health-care expenses are incurred during the acute phase of the ictus and the other 46% during the first year of follow-up. PMID- 10478370 TI - [Perception of long term overload in care-givers of patients who have survived a stroke]. AB - INTRODUCTION: Increased survival of patients with stroke, and reduction in the time spent in hospital, has led to more domiciliary care and extra problems of adjustment for the patient chronically disabled by the stroke and for his carer. MATERIAL AND METHODS: Ninety patient-carer units from the Stroke Unit of the Hospital Universitario San Carlos in Madrid were evaluated one year after the stroke. Eighty carers were assessed on the Zarit Overload Scale, SF-36 Health Questionnaire and the Quality of Life Questionnaire for carers. RESULTS: The carer overload after one year, as measured on the Zarit Scale correlated with the neurological deficit on the Scandinavian Scale (p = 0.003), disability according to the Barthel index (p < 0.0001), female sex (p = 0.0009) and state of mind (p = 0.0001) of the patients. The health SF-36 profile categories most affected were mental health, vitality, pain and general health with average values of 61-62; women scored worse than men in the pain category (p = 0.01) and in physical functions (p = 0.005). Forty two percent recognized that their role was emotionally exhausting and 56% had low morale on the quality of life questionnaire for carers. CONCLUSIONS: Overload of the carers of stroke patients has a considerable effect on their emotional state and social relations, so evaluation is necessary to avoid pathological effects on careers of chronic overload. PMID- 10478372 TI - [Surgery unit for epilepsy]. AB - INTRODUCTION: Only few medically refractory epileptic patients are evaluated for surgical treatment, in spite of the good results obtained at the Centers where epilepsy surgery is performed. OBJECTIVE: We are presenting the way the Gregorio Maranon Hospital Epilepsy Surgery Unit functions and their casuistry. PATIENTS AND METHODS: This Unit is composed by members of the Services of Neurosurgery, Neurology, Neurophysiology, Neuroradiology, Psychology, Neuropediatry, Nuclear Medicine and Neuroanestesiology. Patients are studied by means of protocols. RESULTS: Twenty cases were operated upon. The procedures were temporal lobectomy in 9 cases, amygdalo-hipocampectomy in 4, frontal lobectomy in 3, frontal topectomy in 2, partial hemispherectomy in 1 and vagal nerve stimulation in 1. In 16 of the 19 surgery specimen there were some pathological abnormalities. In the short follow-up 13 patients are seizures free experience significant improvement. PMID- 10478371 TI - [The usefulness of a diagnostic study of mental retardation]. AB - INTRODUCTION: The etiological diagnosis of mental retardation is of interest for several reasons, such as being able to inform parents of the risk of recurrence, and offer the possibility of prenatal diagnosis when possible. It is therefore not surprising that doctors use all means available for this, although the real usefulness of this has not been fully studied. OBJECTIVE: To find the frequency with which an etiological diagnosis is made after relevant investigations. PATIENTS AND METHODS: We describe a retrospective study of persons aged between 0 and 14 years, who were seen by specialists in the hospitals of Gandia and Denia over a period of 7 years. Cases in which the diagnosis was already known or there was isolated motor or language retardation were excluded from the study. RESULTS: An etiological diagnosis was established in 15.4% of the 65 children who fulfilled the criteria of inclusion or exclusion. CONCLUSIONS: The little help obtained from the tests done, and the fact that the number of such tests did not affect the possibility of reaching a diagnosis, made us reconsider the habitual complementary investigations. It would be useful if other centres carried out similar studies so that the results may be compared. PMID- 10478373 TI - [Motor and sensory nerve conduction in patients with carpal tunnel syndrome and diabetic polyneuropathy]. AB - INTRODUCTION: The combination of carpal tunnel syndrome and diabetic polyneuropathy is common, and it is important to establish the correct diagnosis since carpal tunnel syndrome can be successfully treated by surgery, even in diabetic patients. Both conditions have similar clinical features and the usual neurophysiological investigations show very similar results. OBJECTIVE: To determine the differential diagnosis between carpal tunnel syndrome and diabetic polyneuropathyP. PATIENTS AND METHODS: Sensory and motor neuro-conduction studies were done on the median and ulnar nerves of a group of 30 healthy persons (group A), 30 patients with a history of carpal tunnel syndrome (group B) and 30 patients with diabetes mellitus type I or type II (group C) with diabetic polyneuropathy. The physiological variables in which the greatest differences were seen in the three groups were: the speed of sensory conduction in the palm third finger segment of the median nerve, distal latency obtained on stimulation of the fourth finger, distal motor latency of the median nerve and distal latency of the sensory potentials obtained by stimulation at the wrist and recorded at the fourth finger. RESULTS AND CONCLUSIONS: With these variables for prediction, carpal tunnel syndrome was detected in 30% of the patients classified initially on clinical grounds as having diabetic polyneuropathy, and 60% of the patients were correctly reclassified after being initially classified on clinical grounds as having carpal tunnel syndrome. PMID- 10478374 TI - [Tumors of the posterior fossa in children]. AB - INTRODUCTION: Primary tumours of the CNS form 20% of the neoplasias seen in children. They are the second commonest type of cancer seen in childhood, after leukemia. A significant proportion of paediatric cerebral tumours (> 50%) are intratentorial. OBJECTIVE: To find the frequency and clinicopathological behavior of posterior fossa tumours in children seen at our hospital. PATIENTS AND METHODS: From the hospital records, a descriptive, retrospective study was made of 112 children with tumours of the posterior fossa who were attended at the Institute of Neurology and Neurosurgery between January 1980 and December 1997. The variables: age, sex, clinical findings, site and extent of the tumour and its histological description were recorded. The mean, standard deviation, frequency distribution and chi-squared test were used, depending on the type of variable to be analyzed. RESULTS: The age of the patients when the diagnosis was made was between 8 months and 15 years (mean = 8.32 years). The male/female ratio was 59/53 (1.1/1). The commonest site of the tumours was the cerebellum (58.9%). The predominant histological types were medulloblastoma (33.92%) and astrocytoma (19.64%) in the cerebellum, glioma in the brain stem (10.71%) and ependymoma in the i.v. ventricle (8.09%). CONCLUSIONS: Endocranial hypertension and the cerebellar syndrome were the predominant clinical findings in medulloblastoma, cerebellar astrocytoma and ependymoma. In brainstem gliomas there was often involvement of several cranial nerves and long tracts. PMID- 10478375 TI - [Considerations on the concept of epilepsy]. AB - This leading article is written bearing in mind the disparities between different texts and studies on epilepsy. These concern the concept and the definition of epilepsy, although there is now little disparity in the classification of crises and syndromes. We also consider the increasing tendency to include other disorders with their own names and characteristics within the concept of epilepsy. Many of these disorders are also characterized by mental retardation, dementia and neurological deterioration. This contributes to maintain the sombre, ominous concept implicit in the world 'epilepsy' for centuries and is a major factor in the social discrimination of people with epilepsy. We suggest a concept and definition of the disorder differentiating it from epileptic crises and other processes. PMID- 10478376 TI - [Brain stem involvement in eclampsia and HELLP syndrome]. AB - INTRODUCTION: Nowadays, eclampsia is a rare complication of pregnancy and the puerperium. However, it is still one of the main causes of maternal morbi mortality. Systemic findings in eclampsia may sometimes include association with microangiopathic hemolytic anaemia, raised liver enzymes and thrombocytopenia. This clinical combination constitutes the HELLP syndrome (Hemolysis Elevated Liver enzymes Low Platelets). The commonest neurological findings in eclampsia are headache, visual alterations, convulsions and stupor or coma. The presence of clinical symptoms and signs due to brainstem disorders is unusual. CLINICAL CASE: We present the case of a thirty year old woman with eclampsia and associated HELLP syndrome, which briefly affected the brain stem. The patient showed progressive general and neurological improvement. Three months afterwards she still had paresia of the sixth right cranial nerve, due to what was presumed to be a right paramedian infarct of the pons. CONCLUSION: We consider it important not to forget this exceptional association so that suitable treatment may be given to such patients since prognosis is not invariably bad. PMID- 10478377 TI - [Neurological Behcet syndrome. Presentation of three cases]. AB - INTRODUCTION: Behcet's disease is a multisystemic illness in which neurological abnormalities are seen in 5-48% of cases. CLINICAL CASES: We present 3 patients (2 men and 1 women) diagnosed as having Behcet's disease, according to the criteria of the 'International Study Group for Behcet's Disease' of 1990. We describe the clinical findings in these patients, the results of investigations done (MR or CT, evoked potentials-PEV, PESS, PEATC-EEG, ECN, CSF and neuropsychological tests), their course and response to treatment with hyperimmune human gammaglobulim at a dose of 400 mg/kg body weight in 2 patients. All three patients had repeated episodes of meningitis and alterations of the cranial nerves; two patients had epileptic seizures and two had signs of cerebellar disorders and dementia at some time during their illness. One patient died and the other two survived with severe disabilities. Cranial CT showed a cerebral infarct in one patient; evoked potentials showed axon damage in the two patients in whom this was studied. In the only patient in whom ECN was done, signs of axonal polyneuropathy were found. In all three patients there was pleocytosis in the CSF. In the patients in whom hyperimmune human gammaglobulin was given, the results were: one improved after this treatment; in the other there was clinical deterioration, but this was considered to be secondary to a curettage done the day before. CONCLUSION: The presence of neurological findings in Behcet's disease not caused by cerebral vein thrombosis worsens the prognosis of these patients. PMID- 10478378 TI - [Behavior disorders during REM sleep. Two clinical cases]. AB - INTRODUCTION: The behavior disorder occurring during REM (Rapid Eye Movement) sleep is a parasomnia characterized by absence of atonia typical of this phase of sleep, although the other characteristics are maintained, namely rapid eye movements and desynchronization of cortical electrical activity. Clinically it is accompanied by abrupt, often violent movements, which may involve a limb or the trunk in relation to dreams typical of this phase of sleep, and which may interrupt sleep. Many pathological processes have been described, including: the Shy-Dragger syndrome, Parkinson's disease, olivopontocerebellar atrophy, multisystemic atrophy, in relation to certain antidepressant drugs, and most frequently the idiopathic form. CLINICAL CASES: We present two cases, one diagnosed as olivopontocerebellar atrophy and another in which there were no pathological findings. Both were referred to our department for the study of possible sleep disorders. In both cases neurophysiological studies, basically polysomnography with monitorization of various muscle groups and video, led to the diagnosis. CONCLUSIONS: We discuss the diagnostic and physiological criteria, and physiopathological explanations of each case, with special reference to N Methyl-D-Aspartate (NMDA) and non-N-Methyl-D-Aspartate (non-NMDA) receptors. Finally we consider the pharmacological treatment of this disorder. PMID- 10478379 TI - [Peduncular hallucinosis associated with a space occupying lesion of the brain stem]. AB - INTRODUCTION: Traditionally visual hallucinations have been divided into simple and complex. Complex hallucinations due to disorder of the brainstem have been described, and this process referred to as peduncular hallucinosis. CLINICAL CASE: A 74 year old woman with a history of endometrial carcinoma with metastases to bones and lungs, was admitted for progressive ataxia and paraesthesias of the left half of her body. During her admission to hospital she had complex visual hallucinations of cartoon animals, animals, people and geometrical figures. The patient herself realized that they were hallucinations. On cerebral computerized tomography there was a space occupying lesion in the pons-midbrain region of the brainstem. In the clinical context of this particular patient, it was considered to probably be a metastasis from the carcinoma of the endometrium. CONCLUSIONS: Apart from its theoretical interest, peduncular hallucinosis has well defined stereotyped characteristics, and recognition of these may be useful in topographical diagnosis of the lesion. PMID- 10478380 TI - [Dystonic movements: a possible secondary effect of gabapentin]. PMID- 10478381 TI - [Awareness of cerebral vascular disease in the population]. PMID- 10478383 TI - [Additional electrodes on hairy skin in the EEG evaluation of patients with temporal lobe epilepsy]. PMID- 10478382 TI - [Cerebral mucormycosis in a patient with granulocytopenia]. PMID- 10478384 TI - [Etiology and treatment of Chiari I/syringomyelia complex]. PMID- 10478385 TI - [Cranial polyneuritis: a variant of Guillain-Barre syndrome]. PMID- 10478386 TI - [Objective evaluation of muscular elasticity using routine general practice isokinetic test systems]. PMID- 10478387 TI - [Gaining knowledge or recording data by experimental studies?]. PMID- 10478388 TI - [Epidemiology of sports injuries--25-year-analysis of sports orthopedic traumatologic ambulatory care]. AB - In a 25-years period (1972-1997) 30,603 sportsmen, having a total of 34,742 sports related injuries, were treated in our outpatient-department specialized in sports orthopaedics and traumatology. All cases were systematically recorded and analysed right from the beginning. This is the most extensive clinical statistics currently known. The absolutely highest incidence rate of sports-related injuries is seen in common disciplines like soccer with 10,493 (34.3%), skiing with 3632 (11.9%), handball 2307 (7.5%), tennis 1643 (5.4%) and volleyball 1550 (5.1%). 3/4 were male. The distribution of age shows a significant peak between 20 and 29 years of age, whereas in women we found a wide plateau between 10 and 39 years. Compared to earlier investigations we have an increase of injuries in higher age. In 72.4% the lower extremities are mainly effected, followed by the upper extremities (21.8%) and the spine with 3.0%. With increasing tendency knee injuries take main part (36.6%--12,708 cases) followed by injuries of the ankle (19.9%--6920 cases), shoulder (7.7%), lower leg (7.0%) and fingers (5.8%). Main diagnosis were: distortion (32.6%); ruptures of ligaments and menisci (21.5%); fractures (10.5%) and lesions of muscles and tendons (8.8%). Comparing our 15- and 25-years studies we found an interesting significant increase of injuries in skiing, tennis and physical exercising by 25%, in cycling four times. New disciplines like squash, snowboarding, mountainbiking and inlineskating have been added. All together the number of injuries is distributed to 87 different disciplines. In 1998 26.7 million sportsmen (one third of the German population) were member of the German Sports Association (DSB). The number of annual accidents in sports and sporting spare time activities is estimated at about 1.5 2 million, that's 25-30% of all accidents. The analysis of almost 35,000 treated sports injuries and further evaluation of more than 5000 orthopaedic examinations of top athletes are useful for analysis comparing single disciplines. PMID- 10478389 TI - ["Tennis leg": ultrasound differential diagnosis and follow-up]. AB - Tennis leg is a common muscle injury in men older than 40 years participating in racquet sports, alpine skiing and running. It is defined as a partial or total rupture of the medial head of the gastrocnemius muscle which usually is well demonstrable by ultrasound. Main symptom is a sudden sharp pain leading to an obligate immediate stop of sport activity. Clinically pain on local pressure or stretching of the muscle will be reported, local defects can be palpated. 25 patients were prospectively examined by ultrasound according to a new developed grading system. A grade-adapted conservative therapy program led to a wide relief of pain after about two weeks. Full range of sport ability was only regained after at least three weeks. Ultrasonography is able to clearly demonstrate the size and grade of the lesion, thus enabling a well defined therapy program and prognostic conclusions. PMID- 10478390 TI - [Fracture of the pisiform bone in inline skating]. AB - During the last years inline-skating became one of the most familiar kind of sports with more than 12 million inline-skaters. The danger of injuring is often underestimated. Protectors and technique are important for an effective prophylaxis of injury. The upper extremity is mainly endangered. But despite effective protectors there is still the danger of injury. In particular protectors of the hand can shift the force when falling and result in a fracture of the Os pisiforme. This fracture can be missed in standard x-rays and only be diagnosed with special x-rays. PMID- 10478391 TI - [Stress fracture of the ulna in a table tennis player]. AB - This case report describes a stress fracture of the ulna in a 26-year-old professional table-tennis player. X-ray films remained negative so that the diagnostic is based on MRT. This stress fracture is caused by changing the intensity of training and changing the table-tennis-racket. PMID- 10478392 TI - Please put out that cigarette, grandpa. PMID- 10478393 TI - British Columbia's "Tobacco Industry's Poster Child": one part of a bigger picture. British Columbia Ministry of Health and Ministry Responsible for Seniors. PMID- 10478394 TI - Corky, the satellite, and Fishbone Fred. PMID- 10478395 TI - Gutka: a major new tobacco hazard in India. PMID- 10478396 TI - Consensus for tobacco policy among former state legislators using the policy Delphi method. AB - OBJECTIVE: To test a novel approach for building consensus about tobacco control policies among legislators. DESIGN: A pilot study was conducted using a two round, face-to-face policy Delphi method. PARTICIPANTS: Randomly selected sample of 30 former Kentucky legislators (60% participation rate). MAIN OUTCOME MEASURE: Consensus on tobacco control and tobacco farming policies. RESULTS: Former state legislators were more supportive of tobacco control policies than expected, and highly supportive of lessening the state's dependence on tobacco. Former state legislators were in agreement with 43% of the second-round items for which there was no agreement at the first round, demonstrating a striking increase in consensus. With new information from their colleagues, former lawmakers became more supportive of workplace smoking restrictions, limitations on tobacco promotional items, and modest excise tax increases. CONCLUSIONS: The policy Delphi method has the potential for building consensus for tobacco control and tobacco farming policies among state legislators. Tobacco control advocates in other states might consider using the policy Delphi method with policymakers in public and private sectors. PMID- 10478398 TI - Impediments to the enforcement of youth access laws. AB - OBJECTIVE: To recognise obstacles to the implementation of the effective enforcement of tobacco sales laws and to identify measures that could be taken to overcome these obstacles. DESIGN: Interviews were conducted with health department officials in Massachusetts communities to determine why their efforts to prevent illegal sales of tobacco to minors had been only partially successful. SETTING: Urban, suburban, and rural communities in Massachusetts, USA. RESULTS: Organisational problems, court challenges to citations, budgetary threats, and political pressure all combined to reduce the frequency of enforcement inspections to half the intended rate. Political pressure resulted in the exclusion of older youths from compliance tests, further undermining enforcement efficacy. CONCLUSIONS: Suggestions for addressing the problems include obtaining support from the community, keeping elected and court officials informed about plans for merchant education and law enforcement, using efficient enforcement protocols, setting merchant compliance goals, and advising the public and government officials about progress towards those goals, using older youths to make purchase attempts, and testing all merchants frequently. PMID- 10478397 TI - Arizona's tobacco control initiative illustrates the need for continuing oversight by tobacco control advocates. AB - BACKGROUND: In 1994, Arizona voters approved Proposition 200 which increased the tobacco tax and earmarked 23% of the new revenues for tobacco education programmes. OBJECTIVE: To describe the campaign to pass Proposition 200, the legislative debate that followed the passage of the initiative, and the development and implementation of the tobacco control programme. DESIGN: This is a case study. Data were collected through semi-structured interviews with key players in the initiative campaign and in the tobacco education programme, and written records (campaign material, newspapers, memoranda, public records). RESULTS: Despite opposition from the tobacco industry, Arizonans approved an increase in the tobacco tax. At the legislature, health advocates in Arizona successfully fought the tobacco industry attempts to divert the health education funds and pass preemptive legislation. The executive branch limited the scope of the programme to adolescents and pregnant women. It also prevented the programme from attacking the tobacco industry or focusing on secondhand smoke. Health advocates did not put enough pressure at the executive branch to force it to develop a comprehensive tobacco education programme. CONCLUSIONS: It is not enough for health advocates to campaign for an increase in tobacco tax and to protect the funds at the legislature. Tobacco control advocates must closely monitor the development and implementation of tax-funded tobacco education programmes at the administrative level and be willing to press the executive to implement effective programmes. PMID- 10478399 TI - Passive smoking as well as active smoking increases the risk of acute stroke. AB - OBJECTIVE: To estimate the relative risk of stroke associated with exposure to environmental tobacco smoke (ETS, passive smoking) and to estimate the risk of stroke associated with current smoking (active smoking) using the traditional baseline group (never-smokers) and a baseline group that includes lifelong non smokers and long-term (> 10 years) ex-smokers who have not been exposed to ETS. DESIGN AND SETTING: Population-based case-control study in residents of Auckland, New Zealand. SUBJECTS: Cases were obtained from the Auckland stroke study, a population-based register of acute stroke. Controls were obtained from a cross sectional survery of major cardiovascular risk factors measured in the same population. A standard questionnaire was administered to patients and controls by trained nurse interviewers. RESULTS: Information was available for 521 patients with first-ever acute stroke and 1851 community controls aged 35-74 years. After adjusting for potential confounders (age, sex, history of hypertension, heart disease, and diabetes) using logistic regression, exposure to ETS among non smokers and long-term ex-smokers was associated with a significantly increased risk of stroke (odds ratio (OR) = 1.82; 95% confidence interval (95% CI) = 1.34 to 2.49). The risk was significant in men (OR = 2.10; 95% CI = 1.33 to 3.32) and women (OR = 1.66; 95% CI = 1.07 to 2.57). Active smokers had a fourfold risk of stroke compared with people who reported they had never smoked cigarettes (OR = 4.14; 95% CI = 3.04 to 5.63); the risk increased when active smokers were compared with people who had never smoked or had quit smoking more than 10 years earlier and who were not exposed to ETS (OR = 6.33; 95% CI = 4.50 to 8.91). CONCLUSIONS: This study is one of the few to investigate the association between passive smoking and the risk of acute stroke. We found a significantly increased risk of stroke in men and in women. This study also confirms the higher risk of stroke in men and women who smoke cigarettes compared with non-smokers. The stroke risk increases further when those who have been exposed to ETS are excluded from the non-smoking reference group. These findings also suggest that studies investigating the adverse effects of smoking will underestimate the risk if exposure to ETS is not taken into account. PMID- 10478401 TI - An evaluation of a theatre production to encourage non-smoking among elementary age children: 2 Smart 2 Smoke. AB - OBJECTIVE: To evaluate the impact of a theatre production on smoking-related attitudes, norms, and intentions of children in grades 1-6 (aged 6-12 years). DESIGN: Seventeen schools were randomly selected among 160 that were participating in the implementation of the theatre production 2 Smart 2 Smoke. Schools that participated in the theatre production after 3 December 1997 were assigned as control schools. Assignment of schools to a given date for the theatre production was a random process. Students in grades 1-6 were surveyed before and after the theatre production and associated activities. The data were examined for pretest-posttest differences and intervention-control differences. The school was the unit of analysis. SETTING: Elementary schools in the Twin Cities metropolitan area. PARTICIPANTS: Students in grades 1-6 in 17 elementary schools. INTERVENTION: Two plays 2 Smart 2 Smoke for grades 1-3 (6-8 year olds) and grades 4-6 (9-12 year olds), respectively, with follow-up activities for the classroom and home. A national theatre company performed the plays at the schools. MAIN OUTCOME MEASURES: Intention to smoke in the future, normative expectations about how many people smoke, functional meanings of smoking, expected outcomes of smoking. RESULTS: 10% more students reported that they would never smoke a cigarette after the theatre production. Students in grades 4-6 showed changes in the functional meanings and expected outcomes of smoking. Students in grades 1-3 showed changes in normative expectations. CONCLUSIONS: Further research on the impact of live theatre productions as a smoking prevention strategy is recommended. PMID- 10478400 TI - Development of smoking by birth cohort in the adult population in eastern Finland 1972-97. AB - OBJECTIVE: To analyse the dynamics of smoking prevalence, initiation, and cessation in relation to sex, age, birth cohort, study year, and educational level. DESIGN: Six independent cross-sectional population surveys repeated every five years between 1972 and 1997. SETTING: The provinces of North Karelia and Kuopio in eastern Finland. SUBJECTS: Independent random samples of 18,088 men and 19,200 women aged 25-64 years. Those comprising the oldest birth cohort were born in 1913-17 and those in the youngest were born in 1968-72. RESULTS: Among men the prevalence of smoking decreased over time, but the cohort effect observed in smoking initiation was obscured by the changes in smoking cessation. Differences between the educational categories were small. Among women the prevalence of smoking increased during the study period. This was mainly caused by the less highly educated, in whom smoking initiation clearly increased in successive birth cohorts, but a more moderate cohort effect was also present among the more highly educated women. CONCLUSIONS: In men decreased initiation and increased cessation contributed to the downward trend in smoking prevalence, whereas among women, changes in smoking were mostly caused by augmented initiation in successive birth cohorts. During the study period educational inequalities in smoking widened, as the less highly educated came increasingly to form the smoking population. PMID- 10478402 TI - Quit and Win campaigns as a long-term anti-smoking intervention in North Karelia and other parts of Finland. AB - OBJECTIVE: To evaluate Quit and Win campaigns repeated in North Karelia and rest of Finland. DESIGN: Repeated comparisons of participation rates, abstinence rates, and other measures between North Karelia and the rest of Finland. SUBJECTS: Adult daily smokers in Finland participating in the Quit and Win contests in 1986-1997. INTERVENTIONS: Quit and Win smoking cessation campaigns targeted at adult daily smokers throughout Finland in 1986, 1989, 1994, 1996, and 1997, including more intensive activities in North Karelia. MAIN OUTCOME MEASURES: Participation rates, self reported six-month abstinence rates, other effectiveness measures (% of smokers who attended, intended, tried, and succeeded in cessation). RESULTS: North Karelia's participation rates were significantly higher in each campaign compared with the rest of Finland. The abstinence rates in North Karelia were also higher, the difference being significant in 1986 and 1994 (p < 0.05). In the target population in 1996 over 75% of smokers in North Karelia, compared with 40% of smokers surveyed elsewhere, reported awareness of the campaign (p < 0.001). Approximately 9% of the smokers in North Karelia and 6% elsewhere intended to participate (p = NS). Over 2% in North Karelia, compared with less than 1% elsewhere, tried to quit (p < 0.001). Among the targeted group, 0.3% of North Karelian smokers were complete abstainers throughout the 12 months of follow up, compared with an average of 0.1% in other areas (p < 0.001). CONCLUSIONS: The Quit and Win campaign is a feasible cessation method in long term community-wide programmes. Intensified community activities are associated with higher success. In repeat campaigns, high participation and abstinence rates can be maintained. PMID- 10478403 TI - Smoking status by proxy and self report: rate of agreement in different ethnic groups. AB - OBJECTIVE: To examine the rate of agreement between proxy and self report of smoking status in Hispanics compared with other ethnic groups. DESIGN: Data source is the 1990 California Tobacco Survey (CTS) which includes proxy and self reported smoking status. The CTS is a random digit dialed survey conducted in 57,244 households. A sample of 10,011 adults was included in the analyses. MAIN OUTCOME MEASURE: Percentages of agreement and disagreement between self report and report by other member of the household on smoking status. RESULTS: Cohen's kappa coefficients of agreement on smoking status between self report and proxy report was highest in non-Hispanic whites and African Americans (kappa = 0.91), followed by Asian Americans (kappa = 0.82) and Hispanics (kappa = 0.76). Among adults identified as current smokers by proxy, a lower percentage of Hispanics compared with non-Hispanics indicated that they were current smokers (odds ratio (OR) = 3.74, 95% confidence intervals (CI) = 3.28 to 4.20). Furthermore, agreement between proxy and self report was also lower in Hispanics of low acculturation compared with Hispanics with a high level of acculturation (OR = 0.40, 95% CI = 0 to 0.94). CONCLUSIONS: The agreement between self reported and proxy reported smoking status is higher among non-Hispanics compared with Hispanics. Smoking rates in different ethnic groups that are estimated by telephone surveys including proxy and self report might not be comparable. PMID- 10478404 TI - Determinants of smoking behaviour among adolescents in Semarang, Indonesia. AB - OBJECTIVES: To determine the smoking prevalence and to examine the determinants of smoking behaviour among male adolescents in Semarang, Indonesia. DESIGN: A random sample of schools in Semarang (population 1.5 million) was obtained using a stratified sampling procedure (strata based on type of school and district). A total of 149 schools were selected (response rate 72%). Within the schools 186 classes were selected, targeting the 11, 13, 15, and 17 year olds. An anonymous, self administered questionnaire was filled in by all students present at the day of the survey (total sample size 6276). OUTCOME MEASURES: Smoking prevalence, exposure to family and peer smoking, and other variables that may be related to smoking. Logistic regression was used to examine the determinants of smoking behaviour. Only male students were included in the analysis. RESULTS: Smoking increased dramatically between the ages of 11 and 17, from 8.2% to 38.7%. The variance explained by the regression model increased from 19.8% for 11 year olds to 53% for 17 year olds. The smoking behaviour of best friends was the most powerful determinant of smoking, and this was consistent across the age groups. Best friends' attitudes towards smoking and older brothers' smoking behaviour were also important determinants of smoking. CONCLUSIONS: Smoking prevalence among male adolescents in Semarang, Indonesia is high. Effective smoking prevention programmes should take into account the dominant influence of peers in the onset and maintenance of smoking behaviour. In general, school related items had a less important role in predicting smoking behaviour than expected. PMID- 10478405 TI - Smoking prevalence of female nurses in the national hospitals of Japan. AB - OBJECTIVE: To estimate the prevalence of smoking and the attitudes towards the restriction of smoking at work among female nurses in the national hospitals in Japan. DESIGN: Questionnaires mailed to 14 randomly selected national hospitals and sanitariums in Japan in 1993. SUBJECTS: 2207 female nurses. MAIN OUTCOME MEASURES: Smoking status and history, and attitudes towards the restriction of smoking at work. RESULTS: The prevalence of smoking among female nurses was 18.6%, which was higher than the age-adjusted prevalence of the general female population using this study's subjects as a standardised population. Banning smoking in the hospital in which they worked was supported by 15.0%, whereas 81.6% supported the restriction of smoking. CONCLUSIONS: The results showed that smoking is more common among female nurses than among the general female adult population. The survey suggested that nurses favour restriction, but not banning, of smoking. PMID- 10478406 TI - Curbing the epidemic: governments and the economics of tobacco control. The World Bank. PMID- 10478407 TI - Blowing smoke: how cigarette manufacturers argued that nicotine is not addictive. PMID- 10478408 TI - Tobacco Free Initiative, ASH Australia, Brown and Williamson, smoke free children in Ireland, Henley order, smoke signals, and smoking among Hong Kong's children. PMID- 10478409 TI - A girl and her chew: one woman's story of spit tobacco use. PMID- 10478410 TI - The death toll from tobacco: a crime against humanity? PMID- 10478411 TI - The "success" of Philip Morris' campaign on environmental tobacco smoke in The Netherlands. PMID- 10478412 TI - The United States Navy attracts young women who smoke. PMID- 10478413 TI - Eliminating the nicotine in cigarettes. PMID- 10478414 TI - The future of tobacco product regulation and labelling in Europe: implications for the forthcoming European Union directive. AB - The European Commission has announced that it is considering legislation concerning further restrictions on cigarette tar and nicotine yields, as well as new provisions to regulate additives and the labelling of tobacco products. This report considers these issues and their relation to public health. In particular, we argue that further reductions in tar and nicotine yields as measured by the International Standards Organisation/Federal Trade Commission (ISO/FTC) method will be largely cosmetic and certainly misleading to consumers. If a new directive uses the ISO/FTC methodology as a basis for regulation, it risks lending further official support to the concept of "low tar" cigarettes, which may be used by smokers as an alternative to smoking cessation. Although new regulations based on the ISO/FTC methodology may appear to offer health gains, these will be illusory and there may even be negative health consequences, as has been the case with these tests up to the present. We therefore make the following recommendations for the way forward. PMID- 10478415 TI - Ultrastructural study of sheep lacrimal glands. AB - Sheep lacrimal glands are mixed glands, consisting of tubulo-acinar units succeeded by ducts of simple morphology. The secretory portions consist of three cell types: mucous, seromucous and serous, which may be intermingled in the same acinus or may form acini wholly made of only serous or mucous cells. Mucous cells show a rough endoplasmic reticulum that is reduced to a few cisternae located near the cell base and among the interstices of the secretory droplets. Mucous granules appear uniformly electron-lucent. Serous cells display a typical structure; serous granules can be uniformly electron-dense or composed of dense inclusions dispersed in an electron-lucent matrix. The seromucous granules have a bizonal substructure: a dense core is embedded in a highter matrix. Secretory acini are succeeded by intercalated ducts; the epithelium of these ducts gradually increases in height to form a kind of excretory duct, without the intervention of striated ducts. PMID- 10478416 TI - Safety evaluation of the SAG2 rabies virus mutant in Tunisian dogs and several non-target species. AB - The safety of the SAG2 rabies virus, a highly attenuated mutant of the SAD strain intended to vaccinate dogs by the oral route, was evaluated in local Tunisian dogs and in five other local species likely to consume vaccine baits. These species were the domestic cat (Felis catus), the jackal (Canis aureus), the jerboa (Jaculus orientalis), the merion (Meriones sp.) and the gerbil (Gerbillus campestris). The vaccine was administered orally to 21 dogs, 11 cats and eight jackals and orally or intramuscularly to 62 wild rodents of the above-mentioned species. Seven dogs, one cat, five jackals all juvenile and with poor health status) and two rodents died for intercurrent causes. The others were observed for 60-180 days. No animal showed any rabies symptom. Seroneutralizing antibodies were observed in all experimental groups, only after vaccination, with the highest rate being observed in jackals and rodents. The rabies virus was detected in the oral cavity of three cats 6 h after oral instillation, but was not isolated later either in saliva or in salivary glands. Tissue samples (brain and salivary glands) from dead or euthanized animals were examined for the rabies virus antigen by a fluorescent antibody test. No rabies antigen was detected. These trials confirm the safety of the SAG2 strain on the Tunisian species already demonstrated by other authors on many other target and non target species. PMID- 10478417 TI - The effects of injectable sodium selenite on immune function and milk production in Sardinian sheep receiving adequate dietary selenium. AB - The aim of this preliminary study was to determine the effects of selenium (Se) injection on Se status, cell-mediated immunity (CMI), milk yield and milk somatic cell count (MSCC) of ewes fed adequate amounts of Se, and on Se status, passive immunization and CMI of their offspring. Thirty days before lambing, 36 Sardinian ewes were assigned to one of three groups. One group (NT) was not treated; a second group (BL) was given 5 mg of Se on day 30 before lambing; a third group (BLL) was given 2.5 mg of Se on day 30 before lambing and at lambing. Selenium was given intramuscularly as sodium selenite. Selenium status was assessed by measuring glutathione peroxidase activity of erythrocytes (GSHpx-E). The CMI was measured by determining the increases in double skinfold thickness after intradermal injection of phytohaemagglutinin (PHA). Compared to their NT counterparts, ewes belonging to BL and BLL groups and their offspring had significantly higher GSHpx-E (P < 0.01). The GSHpx-E values of lambs were positively related to those of their mothers (P < 0.0005). Ewes of group BL had a greater (P < 0.01) response to PHA 6 h after injection than ewes of the NT group. Lambs born to BL and BLL ewes had a greater (P < 0.0001) response to PHA 24 h after injection. Responses of ewes and lambs to PHA 24 h after injection were positively related (P < 0.05). Serum immunoglobulin at 10 days of age did not differ significantly among the three groups of lambs. Compared to the NT group, milk yield on day 70 of lactation was significantly higher in BL ewes (P < 0.05). The MSCC was not affected significantly by Se injection. Immunoresponsiveness and milk yield might represent additional and appropriate criteria to consider when re-evaluating Se requirements of dairy sheep. PMID- 10478418 TI - Changing reactivity of caprine and ovine mononuclear phagocytes throughout part of the life cycle of Oestrus ovis: assessment through spontaneous and inductible NO production. AB - The in vitro reactivity of monocyte-derived macrophages (MDM) from Oestrus ovis (O. ovis) artificially infested lambs and kids was determined by measuring their production of nitric oxide (NO) during the course of infestation. In both species, crude antigenic preparations obtained from O. ovis first instar larvae (L1) were found to significantly (P < 0.01) inhibit this NO production, whereas O. ovis second instar (L2) extract stimulated it. Furthermore, this NO production by MDM decreased during infestation and was related to blood eosinophilia. It appears that crude antigenic extract from O. ovis modified the NO activity of macrophages from lambs and kids infested with O. ovis larvae. PMID- 10478419 TI - Hyperimmune bovine colostrum treatment of moribund Leopard geckos (Eublepharis macularius) infected with Cryptosporidium sp. AB - Therapy based on the protective passive immunity of hyperimmune bovine colostrum (HBC) was applied to 12 moribund Leopard geckos (Eublepharis macularius) infected with Cryptosporidium sp. The geckos were lethargic and moderately to severely emaciated, weighing on average 36% of the baseline body weight value. Seven gastric HBC treatments at 1-week intervals each decreased the relative output of Cryptosporidium sp. oocysts and the prevalence of oocyst-positive fecal specimens. Histologically, after 8 weeks of therapy, seven out of 12 geckos had only single developmental stages of Cryptosporidium sp. in the intestinal epithelium, and three, one and one geckos had low, moderate and high numbers, respectively, of the pathogen developmental stages. The HBC therapy was efficacious in decreasing the parasite load in moribund geckos. Morphometric and immunologic analysis of Cryptosporidium sp. oocyst isolates originating from Leopard geckos (E. macularius) demonstrated differences between gecko-derived oocyst isolates and isolates of C. serpentis recovered from snakes. PMID- 10478420 TI - Heat inactivation of the neuraminidase and haemagglutinin estimated in the agglutination-separation reactions using red blood cells sensitized with Newcastle disease virus. AB - The agglutination-separation (AS) reactions estimate the effects of heat on the release of altered Newcastle disease virus (NDV) and HN glycoprotein spikes from red blood cells (RBC) sensitized with NDV (SRBC), the inactivations of the neuraminidase (NA), then the haemagglutinin (HA) in a direct assay. Heating SRBC for 1.5 min at 56 degrees C inactivated the NA by 50%; after 4.5 min no separation occurred indicating 100% inactivation of the NA. Heating a suspension of NDV for 78 min inactivated the NA 50% as assayed by cleavage of fetuin. Comparatively, the AS test was up to 52-fold (78 min/1.5 min = 52) more efficient in detecting NA inactivation than was the basic reference test where cleavage of fetuin was assayed. The HA was 50% inactivated after 18 min of heating and 100% inactivated after 36 min as no agglutination was seen. Free HA on SRBC was agglutinated by and thus was titrated with the sialic acid on NRBC. The large area of RBC increased the efficiency of the AS test when compared with tests using suspensions of NDV. At 51-60 degrees C all NA and HA inactivations were sequential, and invariably the NA was more heat labile than the HA. The release of altered NDV and HN spikes was inhibited with mild heat although the separation of SRBC and NRBC continued. Biological purifications showed that the heat stability of the HA and the lability of the NA were genetically stable. PMID- 10478421 TI - Cross-reactive antigens of Haemonchus contortus adult worms in Teladorsagia circumcincta infected goats. AB - The adult Haemonchus contortus somatic antigens responsible for cross-reactivity have been analysed using serum samples from goat kids infected and reinfected with Teladorsagia circumcincta. Goat kids infected with T. circumcincta had similar serum ELISA values against somatic antigens of H. contortus as goats infected with H. contortus itself. Immunoblotting confirmed this extensive cross reactivity particularly in the molecular weight range 105-29 kDa. However, peptides with high (195, 152 and 119 kDa) or low (23 kDa) molecular weight were only faintly recognized by heterologous sera. PMID- 10478422 TI - Effect of field capture on the measurement of cellular immune responses in wild ferrets (Mustela furo), vectors of bovine tuberculosis in New Zealand. AB - Ferrets are recognised as significant wildlife vectors of bovine tuberculosis (Tb) in New Zealand. Disease management strategies, such as the development of a protective wildlife vaccine, could be assisted by the ability to measure pertinent cellular immune responses among wild animals. In the present study, we investigated whether it is possible to measure in vitro lymphocyte reactivity in wild-caught ferrets, and also determined levels of physiological stress in these animals, and we compared these responses to those observed in laboratory maintained domesticated ferrets. Over a 12-month period, 80 ferrets were live captured from a Tb-endemic region (Otago, southern New Zealand); cardiac blood was withdrawn on-site, and mononuclear cell cultures were successfully established from 75 of these animals. Lymphocyte transformation (LT) responses to T cell and T/B cell mitogens (Concanavalin A [Con A] and pokeweed mitogen) were measured via uridine incorporation assay. The magnitude of these responses did not differ significantly between animals that had been captured in wire-framed cage traps and those captured using soft-jawed leg-hold traps. Levels of serum cortisol and glucose (as indicators of physiological and oxidative stress, respectively) were highest in animals captured using leg-hold traps. In comparison to domesticated ferrets, wild-caught ferrets had lower overall LT responses to Con A, but significantly higher levels of serum cortisol. Finally, 10/80 animals captured from the wild were severely diseased (Tb+), as evidenced by gross tuberculous lesions at autopsy. Successful mononuclear cell cultures were established from nine of these animals; LT responses to Con A were significantly lower in Tb+ ferrets than in either wild-caught/non-diseased (Tb-) or domesticated ferrets. These results demonstrate that it is possible to measure cellular immune responses from the blood of wild-caught ferrets, but that field capture and disease status may have detrimental effects on in vitro T cell function, possibly due to the influence of physiological stress. PMID- 10478423 TI - Effects of dimerized lysozyme (KLP-602) on the cellular and humoral defence mechanisms in sheatfish (Silurus glanis): in vitro and in vivo study. AB - This study examined the effects of the dimerized lysozyme (KLP-602) on the immunocompetence cell activity in sheatfish (Silurus glanis) and its influence in vivo on the non-specific defence mechanisms and protection against motile aeromonad septicaemia (MAS). The in vitro study showed that the lysozyme dimer (KLP-602), at concentrations between 5 and 50 micrograms/mL of medium significantly (P < 0.05) increased the respiratory burst activity and potential killing activity of pronephric macrophages, as well as the proliferative ability of pronephric lymphocytes stimulated by ConA and LPS. The in vivo study showed that injecting lysozyme dimer (Lydium-KLP) intraperitoneally at doses of 50 micrograms/kg bw stimulated cell-mediated and humoral-mediated imunity. On day 5, after application of Lydium-KLP in vivo, a statistically higher (P < 0.05) respiratory burst activity and potential killing activity of blood and pronephros phagocytes were observed. A higher proliferative ability of blood and pronephros lymphocytes stimulated by Concanavaline A (ConA) or lipopolysaccharide (LPS) was also observed. At the same time, the myeloperoxidase activity in the PMN cells and the lysozyme activity and total Ig levels in serum were significantly higher (P < 0.05), compared to the control group. A challenge test with Aeromonas hydrophila showed that dimerized lysozyme increased the protection against MAS. Dimerized lysozyme stimulates non-specific cellular and humoral mechanisms and protection against MAS in sheatfish. PMID- 10478424 TI - [Effect of the number of health meals before an infectious meal on the vectorial competence of Glossina morsitans morsitans infected by Trypanosoma congolense IL 1180]. AB - The purpose of this work was to assess the influence of several healthy meals (0, 1 and 2) prior to the infectious one on the vectorial competence of Glossina morsitans morsitans (Mall). The teneral flies (< 32 h old) of this line were divided into three groups. The tsetse flies of group A received no meal. The ones of group B received one healthy meal on day 1, whereas those from group C were given two consecutive healthy meals on days 1 and 2. All the flies were experimentally infected with Trypanosoma congolense IL 1180 when the maximum age reached 32 h for flies with no meal, 56 h for those with one healthy meal and 80 h for those who received two healthy meals. When both sexes were considered, the meso-procyclic and metacyclic indexes as well as the vectorial competence (VC) of the flies receiving no meal were 0.99 +/- 0.01, 0.96 +/- 0.02 and 0.95 +/- 0.03. Considering the flies which were fed one healthy meal, the respective values were 0.42 +/- 0.13, 0.50 +/- 0.01 and 0.21 +/- 0.06, whereas the values for the flies receiving two healthy meals were 0.45 +/- 0.11, 0.29 +/- 0.19 and 0.13 +/- 0.05. The meso-procyclic and metacyclic indexes as well as the VC in both sexes were more important in the flies which received no meal than those fed with one or two healthy meals. The meso-procyclic and metacyclic indexes and VC did not show any significant differences between the flies fed one or two healthy meals, whereas the metacyclic index of male flies which received one healthy meal was significantly higher than those fed two healthy meals. These results indicate that the number of non-infected (healthy) meals prior to an infected meal reduces the interaction between G. m. morsitans infected and T. congolense. PMID- 10478425 TI - B-cell epitopes recognized by Chinese water buffaloes (Bos buffelus) on the 22 kDa tegumental membrane-associated antigen (Sj-22) of the Asiatic bloodfluke, Schistosoma japonicum. AB - The 22.6 kDa tegumental membrane-associated antigen of schistosomes is of recognized importance in immunity to schistosomiasis. In China, bovines are known to play an important role in the transmission of Schistosoma japonicum. Ten buffaloes (Bos buffelus) were vaccinated with a recombinant form (reSj-22) of the S. japonicum 22.6 kDa tegumental antigen (Sj-22) and the sera were used to identify and map possible linear B-cell epitopes on this molecule using a series of 18 overlapping synthetic peptides (P1-P18). Sera from all of the ten vaccinated buffaloes reacted strongly with Sj-22 in western blots and in ELISA, while sera from a further ten adjuvant (Quil A) control buffaloes did not. Four peptides (P3, P8, P9 and P10) were predominantly recognized by at least 90% of the buffalo sera. This pattern of recognition is similar to that obtained in a previous study we undertook in mice immunized with the same antigen whereby peptides 3, 8, 9 and 10 were recognized by over 80% of CBA strain mice. The peptide most frequently recognized by mice (peptide 6), and mapping to an EF-hand calcium binding domain, was recognized by six of the ten vaccinated buffaloes. The major difference between buffaloes and mice occurred with peptide 1 which was recognized very frequently by all three strains of mice tested but was only weakly recognized by three of the ten buffaloes. This study provides a valuable reference for further study on the immunity stimulated by the 22.6 kDa tegumental antigen in the murine model and a natural bovine host of Schistosomiasis japonica. PMID- 10478426 TI - [Analysis based medical training therapy for the spine (FPZ concept): quality assurance in the scope of evidence-based medicine]. PMID- 10478427 TI - [Physical therapy with analgesic effects in inflammatory rheumatic diseases]. AB - In praxis, one is not only concerned about the linear connection between rheumatic inflammation and pain therapy--pharmacological or physical treatment- but about an optimal connection of the whole spectrum of symptoms regarding the joint status with different options of actions on one hand, and the treatment procedure of choice beside pain therapy on the other hand. The understanding of the mode of action regarding physiotherapeutic methods with analgetic effects is due to the knowledge about the pathophysiologic relationship between inflammation and pain. On one hand, the information of pain from the periphery causes a sensibilisation of central nervous areas and therefore allodynia as well as irradiation of the painful area. On the other hand, biochemical processes lead to a neurogenic and alogenic inflammation due to the impact on the pain nerves. Beside the arthritis itself, the physiotherapy of a rheumatic-inflamed painful joint has to take into account the components arthralgia, irritation of the capsula, periarthropathia, and allogenic inflammation. Physiotherapeutic pain treatment has three modes of action: a causal effect on the structural level, suppression of over-excitation of the tissue as well as the central nervous transmission areas, and activation of the decelerating pain modulating systems. Each treatment procedure with analgetic effects like cooling, heating, localized mechanical methods, the activation techniques with proprio- and exteroceptive effects (TENS, Maitland-mobilisation), the electrotherapeutic option, and physical training therapy is characterized with its specific mode of action. The dosage of physiotherapeutic techniques has to be taken into account since it has to remain within physiologic limits and the tissue's irritability must not be reached. PMID- 10478428 TI - [Preconditions for disease-specific exercise therapy]. AB - The exercise therapy is a therapy of physiological reactions. Moving impulses can lead to direct or indirect functional reactions in different systems (i.e. neuromotor, cardiovascular etc.). As an aim within the therapy, exercise is used either under adaptive substituting or compensatory relieving. In order to reach economic and effective functional results, many aspects have to be considered many aspects, such as executing-, planning- and control criteria. They are demonstrated by means of selected samples of diseases and injuries of the locomotor apparatus. PMID- 10478429 TI - [Thermotherapy in rheumatic diseases]. AB - Heat and cold treatment are often used in the physical therapy of rheumatic diseases. Thermotherapy influences perfusion, muscular tone, pain, and inflamed arthritic joints. Cold applications are preferable in treating active arthritis. Heat treatment is advantageous in the management of less active arthritis and non inflammatory rheumatic disorders. Current investigations demonstrate that thermotherapy influences the endocrine--an immune systems. PMID- 10478430 TI - [Occupational therapy in the diseases of the locomotor system]. AB - The spectrum of occupational therapy (OT) services in rheumatology includes the restoration of disturbed muscle function, psychosocial aspects, instruction and training in joint protection and manufacturing of orthotic devices as well as advice and training with technical aids and adapting the home and workplace according to the patient's disability. OT is especially indicated in inflammatory rheumatic diseases with polyarticular involvement, but also in degenerative joint diseases, various forms of soft tissue rheumatism and spinal disorders. It should be prescribed by a physician and functional therapist together with regard to the specific location of functional derangement and is to be dosed individually according to the work-load applied. A wide range of techniques involving different materials--including textiles, leather, wood, clay--enable individual joints or functional entities to be exercised specifically. Integration and coordination of OT with the other elements of multidisciplinary therapy, e.g. pharmacological treatment, physiotherapy, orthopedic surgery is the important task of the rehabilitation team. The objective measurement of therapeutic effects serve the purposes of demonstrating the quality of the results and also of motivating the patient. The methods used will depend on the clinical picture, the structures damaged, and the time available for the therapist. PMID- 10478431 TI - [Effectiveness of back-school programs in controlled trials. A review of the literature]. AB - Backschools are utilized for primary, secondary and tertiary prevention of nonspecific low back pain while there is considerable doubt concerning their effectiveness. We present the results of controlled clinical trials that examine the effectiveness of back schools as reported in recent systematic reviews of the literature. An extensive literature search for systematic reviews summarizing the results of controlled trials was performed. Publications which satisfied a number of inclusion criteria concerning methodology and content were critically appraised. Their main results and conclusions are presented and discussed. PMID- 10478432 TI - [Current status and perspectives of rehabilitation of musculoskeletal diseases]. AB - Musculoskeletal diseases play a prominent role within the system of rehabilitation due to their high prevalence, the often progressive course they take, and the high amount of disabilities caused by them in occupational and everyday activities. In Germany, almost 50% of about 1 million cases of in patient rehabilitation per year belong to this diagnostic group. At present, the repercussions of recent changes in legislation and the resulting decrease by about 30% in the use of rehabilitation have considerable influences on the system of in-patient rehabilitation. In addition, rehabilitation hospitals are exposed to a growing competition by the development of out-patient rehabilitation facilities. The present article describes the actual situation, analyses central dimensions of the process of rehabilitation in musculoskeletal diseases as well as the evidence on the efficacy of current rehabilitation programmes, and then informs on the systematic activities aiming at quality assurance in rehabilitation. PMID- 10478433 TI - [Smoking: health care and politics in flux]. AB - The cigarette is the only legally sold product with carcinogenic, cardiovascular damaging and addictive effects. With a yearly profit of several billions, the tobacco industry supports politicians to solve their tasks, whereas these polticians, do not promote projects against smoking and the protection of nonsmokers, despite of some less compulsory statements. Up to now, the tobacco industry denies the health hazard effects occurring to cigarette smokers after two or three decades. On the other hand, the public health insurance do not provide any financial support for smoking prophylaxis and smoking cessation. Instead of this, they have to cover costs for subsequent health injuries, early disability included, which are more than 100-1000 times higher. The solidary community has to finance the pensions for relatives of those numerous smokers, who died early, and whose lives were shortended by provable 5 to 6 years. Politicians refuse to represent the will of a population majority for measures to prevent nonsmokers. PMID- 10478435 TI - [Guideline report on bronchial asthma. 4: Structured guidelines--Abstracts (USA)]. PMID- 10478434 TI - [The problem of evidence-based guidelines using the example of the treatment of obesity]. AB - Despite many therapeutic strategies used by experts and general practitioners, the obesity problem has still increased in the developed countries. Therefore it is necessary to provide evidence-based references for good clinical practice. The guideline presented in Cologne in 1997 was published in an improved form in 1998. However, the consequences and circumstances do not only promise benefit but lead to new questions: Where are the indications for implementation and the participation of the main users, the general practitioners? Why should the procedures of tests and consensus not used additionally? The co-morbidity, biography, genetics, and the number of relapses of patients are probably not emphasized enough, the use of drugs could be considered more carefully. Although the reviewed guideline is important, there is a lack in providing the information and in advice for procedures of co-operation in primary health care. The great therapeutic problem and understanding for the daily practice has not been estimated enough. PMID- 10478436 TI - [Observations of play and verbal behavior of boys with fragile X syndrome in early childhood]. AB - Reports on development and behaviour in boys with fragile-X syndrome support the idea of a characteristic behavioural phenotype in this special population. Preliminary results are presented for 10 boys with fragile-X syndrome in early childhood. Severe mental handicaps and communicative abnormalities are observed less frequently than was expected on the basis of results reported for school-age children or adults. Boys with fragile-X syndrome show goal-directed and cooperative play behaviours in a Montessori play session, but less persistence and organisation than children with normal development or a mental handicap of heterogeneous origin. Results confirmed these behavioural differences as characteristic aspects of a "behavioural phenotype" in children who already in early childhood have fragile-X syndrome. PMID- 10478437 TI - [Applicability of the Child Behavior Checklist in developmentally delayed children]. AB - OBJECTIVES: The Child Behavior Checklist is used to assess behavioral problems in a large unselected sample of children. In the present study we assess the usefulness of the CBCL in the evaluation of the typical behavioral problems usually reported in children with developmental disorders. METHODS: We examined two groups of children which both had language or communication problems. The first group consisted of 34 children with infantile autism, the second group consisted of 34 age-, sex- and IQ-matched children with a specific developmental speech and language disorder. The CBCL was filled out by the parents in both groups as part of the routine diagnostic procedures. RESULTS: Half of the language-impaired children have Total Behavioral Problem scores within the clinical range. Problems are mainly reported on the scales: "Attention Problems", "Social Problems" and "Withdrawn". Two-thirds of the autistic children have deviant scores on the syndrome scales mentioned above. 32 out of 34 autistic children score within the clinical range on the scale "Thought Problems", whereas only one language-impaired child does so. Single item analysis shows a high prevalence of developmental problems (speech problems, enuresis ...) in both groups. CONCLUSION: The CBCL records characteristic behavioral problems in children with developmental disorders. The problematic behaviors are shown on the syndrome scale level as well as on the single item level. Children with developmental disorders and high scores on the "Thought Problems" scale of the CBCL should be evaluated for the presence of a possible pervasive developmental disorder. PMID- 10478438 TI - [Behavior therapy methods in treatment of schizophrenic adolescents]. PMID- 10478439 TI - [100 years research in reading and spelling--what do we know today?]. PMID- 10478440 TI - [Relapses in the course of anorexia nervosa]. PMID- 10478441 TI - [Differential "conversion disorder" diagnosis]. PMID- 10478442 TI - A mixture of organisms affects cholesterol metabolism together with rat cecal flora. AB - The effects of a mixture of organisms on cecal fermentation and cholesterol metabolism in sham-operated and cecectomized rats were investigated. Male F344 rats, allocated into four groups: cecectomized rats fed a mixture of organisms (CEMO), cecectomized rats fed rice bran (CERB), sham-operated rats fed a mixture of organisms (SHMO), and sham-operated rats fed rice bran (SHRB) for 4 weeks. The diets had 0.5% cholesterol and 0.125% sodium cholate added. There were no significant differences in the body weight gain and food intake among the groups. The cecal pH in the SHMO group was significantly lower than that in the other groups. The total cholesterol and (VLDL + IDL + LDL)-cholesterol concentrations in serum were significantly lower in the SHMO group than that in the SHRB group, and the triacylglycerol concentration in the sham-operated rats tended to decrease compared to the cecectomized rats. The fecal cholesterol excretion in the CERB group was higher than that in the other groups, and that in the SHMO group was significantly higher than in the SHRB group. The acetic acid, propionic acid, n-butyric acid, and total short-chain fatty acid concentrations in the cecum contents were significantly higher in the SHMO group than those in the other groups. Streptococcus, Bifidobacterium, and Lactobacillus in the SHMO group tended to be higher than the other groups and Bacteroidaceae in the CEMO and CERB groups were significantly higher than that in the SHMO group. The results demonstrate that the mixture of organisms was fermented with the cecal contents and that the metabolites such as short-chain fatty acid lowered the serum total cholesterol and liver cholesterol concentrations in the rats fed a cholesterol containing diet. PMID- 10478443 TI - Identification of a member of the serralysin family isolated from a psychrotrophic bacterium, Pseudomonas fluorescens 114. AB - An extracellular metalloprotease named No. 114 protease is one of the major secretions of a psychrotrophic bacterium, Pseudomonas fluorescens 114, the cold adaptation mechanism of which has not been identified. In this study, we purified and cloned No. 114 protease, which is a single polypeptide having a molecular mass of 47 kDa. This protease contains a zinc-binding motif (HEXXHXUGUXH: X, arbitrary amino acid; U, bulky hydrophobic amino acid), glycine-rich repeats (GGXGXD) and no cysteine residue, which are the features specifically found in serralysin subfamily. No. 114 protease has its maximum activity at the temperature of 35-40 degrees C, which is about 20 degrees C lower than that of a serralysin from a mesophilic bacterium, Pseudomonas aeruginosa. All these results imply that No. 114 protease from this psychrophilic bacterium is a unique member of the serralysin group characterized by a low optimal temperature. PMID- 10478444 TI - Kinetic studies on the function of all the conserved tryptophans involved inside and outside the QW motifs of squalene-hopene cyclase: stabilizing effect of the protein structure against thermal denaturation. AB - Site-directed mutagenesis experiments were carried out to identify the responsibility of the eight QW motifs for the reaction catalyzed by squalene hopene cyclase (SHC). Alterations of the conserved tryptophans, which are responsible for the stacking structure with glutamine, into aliphatic amino acids gave a significantly lower temperature for the catalytic optimum as for the mutageneses of QW motifs 4, 5a and 5b, which are specifically present in SHCs. However, there was no change in the optimal temperatures of the mutated SHCs targeted at the other five motifs 1, 2, 3, 5c and 6. Thus, reinforcement against heat denaturation can be proposed as a function of the three QW motifs 4, 5a and 5b, but no function could be identified for the QW motifs 1, 2, 3, 5c and 6, although they are commonly found in all the families of prokaryotic SHCs and eukaryotic oxidosqualene cyclases. On the other hand, the three conserved tryptophans of W169, W312 and W489, which are located inside the putative central cavity and outside the QW motifs, were identified as components of the active sites, but also had a function against thermal denaturation. The other two tryptophan residues of W142 and W558, which are located outside the QW motifs, were found not to be active sites, but also had a role for stabilizing the protein structure. It is noteworthy that the mutants replaced by phenylalanine had higher temperatures for the catalytic optimum than those replaced by aliphatic amino acids. The catalytic optimal pH values for all the mutants remained unchanged with an identical value of 6.0. PMID- 10478445 TI - Effects of Maillard reaction products on the oxidative cleavage and polymerization of protein under ascorbic acid-transition metal system. AB - This study investigated the effects of Maillard reaction products (MRPs) on the oxidative cleavage and polymerization of BSA (bovine serum albumin) in an aqueous system. In L-ascorbic acid (AsA) and Cu(II) or Fe(III) reaction system, 50-60% of BSA was cleaved under physiological conditions (37 degrees C, pH 7.2). The oxidative cleavage induced by AsA-Cu(II) system was suppressed to the extent of 32-86% by model melanoidins or brown pigments from amino acids and foodstuffs. In the AsA-Fe(III) system, the oxidative cleavage was inhibited to the extent of 45 93% by melanoidins and brown pigments. However, this cleavage was promoted by amino acid Amadori rearrangement products and brown pigment from soy paste. Therefore, MRPs show both suppression and promotion activity on oxidative cleavage of BSA in the system of AsA and a transition metal. The quantity of Amadori rearrangement moiety (ARM) in melanoidins from Lysine and brown pigments molecules from foods was also measured. From these data, it was estimated that the suppression and/or promotion of oxidative cleavage of BSA did not only depend on the quantity of ARM, but also depended on the chemical structure of ARM in melanoidins or brown pigments. PMID- 10478446 TI - In vitro anti-human immunodeficiency virus activity of polysaccharide from Rhizophora mucronata Poir. AB - A polysaccharide was extracted with 1% sodium carbonate from the bark of Rhizophora mucronata and its antiviral activities against human immunodeficiency virus (HIV) were assessed by an in vitro cell culture system. The anti-HIV activity of the alkaline extract was mainly recovered in the 25-75% ethanol precipitated fraction. Rhizophora mucronata polysaccharide (RMP) protected MT-4 cells from the HIV-induced cytopathogenicity and blocked the expression of HIV antigens. RMP completely inhibited the viral binding to the cell and the formation of syncytium upon cocultivation of MOLT-4/HIV-1IIIB cells and MOLT-4 cells. These results suggest that RMP inhibited early steps of the virus life cycle especially virus adsorption to the cell. PMID- 10478447 TI - Stimulative effect of a casein hydrolysate on exocrine pancreatic secretion that is independent of luminal trypsin inhibitory activity in rats. AB - We have previously demonstrated that proteins could stimulate pancreatic secretion independently of luminal bile-pancreatic juice (BPJ) in a BPJ-diverted rat. To determine whether luminal protease-independent pancreatic secretion occurs in normal rats with BPJ returned to the upper small intestine, we investigated the pancreatic secretory response to intraduodenal instillation of a casein hydrolysate or the synthetic trypsin inhibitor, FOY 305, at concentrations which could almost equally inhibit hydrolysis of the synthetic substrate for trypsin with the luminal content. FOY 305 at 10 micrograms/ml and casein hydrolysate solutions at both 100 and 200 mg/ml similarly inhibited approx. 80% of the tryptic activity in the luminal contents of the proximal small intestine. Intraduodenal administration of casein hydrolysate solutions (100 and 200 mg/ml) significantly increased pancreatic secretion in a dose-dependent manner. However, intraduodenal administration of FOY 305 (10 micrograms/ml) was ineffective for stimulating pancreatic secretion. These results demonstrate that dietary protein enhances pancreatic secretion independently of the masking of luminal trypsin activity in rats. PMID- 10478448 TI - Accelerated evolution in the protein-coding region of galectin cDNAs, congerin I and congerin II, from skin mucus of conger eel (Conger myriaster). AB - Two cDNAs encoding galectins named congerins I and II from the skin mucus of conger eel (Conger myriaster) were isolated and sequenced. Comparison of the nucleotide sequences of congerins I and II showed that the sequence similarities of the 5' and 3' untranslated regions (86 and 88%, respectively) were much higher than those of the protein-coding region (73%). The numbers of nucleotide substitutions per site (KN) for the untranslated regions are smaller than the numbers of nucleotide substitutions per synonymous site (KS) for the protein coding region. Furthermore, nonsynonymous nucleotide substitutions have accelerated more frequently than synonymous nucleotide substitutions in the protein coding region (KA/KS = 2.57). These results suggest that accelerated substitutions have occurred in the protein-coding regions of galectin genes to generate diverse galectins with different molecular properties. Northern blot analysis showed that both congerins were expressed not only in the skin tissues but also in the stomach of conger eel. PMID- 10478449 TI - Isolation and characterization of the glyceraldehyde-3-phosphate dehydrogenase gene of Lentinus edodes. AB - The glyceraldehyde-3-phosphate dehydrogenase (GPD) gene of Lentinus edodes was isolated from a genomic DNA library and cDNA corresponding to this gene was isolated from a mycelium cDNA library. The L. edodes GPD gene was found to encode a 337-aa protein. By comparison of the cDNA and genomic DNA sequences, the presence of eight introns in the GPD gene was confirmed. The putative amino acid sequence of the L. edodes GPD gene product showed high similarity to those of other basidiomycetes. The results of Southern blot analyses suggested that only one copy of the GPD gene is present in the genome of L. edodes. The promoter region was found to contain a CT-rich stretch, two CAAT boxes and a consensus TATA box. In addition, the transcript of the GPD gene was found to be expressed constitutively and strongly. These results suggest that the promoter of the L. edodes GPD gene may be very useful as a component of transformation vectors. PMID- 10478450 TI - Activation of macrophages and neutrophils by an endothelium growth suppressing factor. AB - An endothelial cell growth-suppressing factor (EGSF) was purified from the serum free conditioned medium of the mouse P388D1 culture in the presence of carboxymethylated curdlan. The purified EGSF showed two bands corresponding to the molecular masses of 55 and 63 kDa by silver staining on a SDS-polyacrylamide gel under reducing conditions. This factor strongly suppressed the proliferation of endothelial cells from bovine artery, human umbilical vein, and human dermal vas capillare and this suppression was observed to be reversible. We found that EGSF was a potent chemoattractant for macrophages and neutrophils. EGSF mediated the adhesion of neutrophils to BAEs and transendothelial migration of neutrophils. Macrophages stimulated by EGSF produced nitrite in a dose-dependent manner. EGSF did not affect the proliferation of T lymphocytes. These findings suggest that EGSF acts not only as a potent inhibitor for the growth of endothelial cells but also an activator for macrophages and neutrophils. Thus EGSF plays a role in an inflammatory response in the endothelium. PMID- 10478451 TI - Muscle hypertrophy in rats fed on a buckwheat protein extract. AB - Growing rats were examined for the influence of a buckwheat protein diet on muscle weight and protein. In experiment 1, the rats were fed on a diet containing either casein or a buckwheat protein extract (BWPE) as the protein source (10%, 20% or 30%) for 5 wk. The relative weights (g per kg of body wt) of the gastrocnemius, plantaris and soleus muscles were higher in the BWPE-fed animals than in the casein-fed ones, but were unaffected by the dietary level of protein. These differences were not associated with growth. In experiment 2, the rats were fed on either a casein or BWPE diet at the 20% protein level for 5 wk. BWPE intake significantly elevated the gastrocnemius muscle weight, carcass protein and water, and reduced carcass fat. These results demonstrate that BWPE consumption causes muscle hypertrophy, elevates carcass protein and water, and reduces body fat. PMID- 10478452 TI - High throughput assay to detect compounds that enhance the proton permeability of Candida albicans membranes. AB - We describe a 96-well microtiter plate format assay to detect changes in proton permeability in membranes of the pathogenic yeast, Candida albicans. Candida albicans cells were incubated with the lipophilic ester of 2',7'-bis-(2 carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF), a pH-sensitive fluorescein derivative. Inside the cells, BCECF was released and trapped in the vacuole. Compounds that destroyed membrane integrity increased the pH value of the vacuole due to proton leakage into the cytoplasm. This was paralleled by an increase in BCECF fluorescence intensity, which could be quantified. The test assay was validated with amphotericin B, as well as with other membrane-active compounds known to increase membrane permeability. Possible applications and limitations of this assay in the field of antifungal drug discovery are discussed. PMID- 10478453 TI - Purification, characterization and gene analysis of N-acetylglucosaminidase from Enterobacter sp. G-1. AB - Enterobacter sp. G-1 is a bacterium isolated previously as a chitinase-producing bacterium. We found this bacterium also produced N-acetylglucosaminidase and characterized that in this study. Extracellular N-acetylglucosaminidase of 92.0 kDa was purified near homogeneity by 8.57-fold from Enterobacter sp. G-1. The optimum temperature and the optimum pH of the purified N-acetylglucosaminidase was 45 degrees C and 6.0, respectively. The N-terminal amino acid sequence of 23 residues of N-acetylglucosaminidase was identified. Based on the N-terminal sequence, we amplified pieces of the DNA fragments by PCR. Using these PCR products as probes, we screened the genomic library and successfully isolated the entire N-acetylglucosaminidase gene (designated nag1) from Enterobacter sp. G-1. The nucleotide sequence of the nag1 gene was found to consist of 2,655 bp encoding a protein of 885 amino acid residues. Comparison of the deduced amino acid sequence from the nag1 gene found 97.3% identity with chitobiase from Serratia marcescens, 54.4% identity with N,N'-diacetylchitobiase from Vibrio harveyi, and 42.7% identity with N-acetylglucosaminidase (ExoI) from Vibrio furnissii. Enzymatic activity assay of N-acetylglucosaminidase indicated stronger activity toward PNP-GlcNAc than PNP-(GlcNAc)2 or PNP-(GlcNAc)3. PMID- 10478454 TI - Molecular mechanism for pore-formation in lipid membranes by the hemolytic lectin CEL-III from marine invertebrate Cucumaria echinata. AB - The pore-forming activity of CEL-III, a Gal/GalNAc specific lectin from the Holothuroidea Cucumaria echinata, was examined using artificial lipid membranes as a model system of erythrocyte membrane. The carboxyfluorescein (CF)-leakage studies clearly indicated that CEL-III induced the formation of pores in the dipalmitoyl phosphatidyl choline (DPPC)-lactosyl ceramide (LacCer) liposomes effectively but not in the DPPC-glucosyl ceramide (GlcCer) liposomes or DPPC liposomes. Such a leakage of CF was strongly inhibited by lactose, a potent inhibitor of CEL-III, suggesting that the leakage is mediated through the specific binding of CEL-III to the carbohydrate chains on the surface of the liposomes. The leakage of CF from the DPPC-lactosyl ceramide liposomes was pH dependent, and it increased with increasing pH. The immunoblotting analysis and circular dichroism data indicated that upon interaction with liposomes, CEL-III associated to form an oligomer concomitantly with a marked conformational change. Furthermore, channel measurements showed that CEL-III has an ability to form small ion channels in the planar lipid bilayers consisting of diphytanoylphosphatidylcholine and human globoside (Gb4Cer)/LacCer. PMID- 10478455 TI - Conformational state of disulfide-reduced ovalbumin at acidic pH. AB - Ovalbumin assumes a highly ordered molten-globule conformation at pH 2.2. To investigate whether or not such structural nature is related to the existence of an intrachain native disulfide bond, the structural characteristics of disulfide reduced ovalbumin at the acidic pH were compared with those of the native disulfide-intact protein by a variety of analytical approaches. The disulfide reduced protein was found to assume a partially denatured molten globule-like conformation similar to the disulfide-intact counterpart as analyzed by the CD and intrinsic tryptophan fluorescence spectra and by the binding of a hydrophobic probe of anilino-1-naphthalene-8-sulfonate. The results from size-exclusion chromatography also showed that the disulfide-reduced and disulfide-intact proteins have essentially the same compact, native-like hydrodynamic volume. The disulfide-reduced protein was, however, highly sensitive to proteolysis by pepsin at the acidic pH under the proteolytic conditions in which the disulfide-intact protein was almost completely resistant. Furthermore, on a differential scanning calorimeter analysis the disulfide-reduced protein had an endothermic transition at a much lower temperature (Tm = 48.5 degrees C) than the disulfide-intact protein (Tm = 57.2 degrees C). Taken together, we concluded that the intrachain disulfide bond should not be directly related to the highly ordered molten globule conformation of ovalbumin, but that its conformational stability depends on the presence of the disulfide bond. PMID- 10478456 TI - Expression of mature pokeweed antiviral protein with or without C-terminal extrapeptide in Escherichia coli as a fusion with maltose-binding protein. AB - Genomic clones encoding the mature pokeweed antiviral protein with or without C terminal extrapeptide (PAPMC and PAPM), which have been reported to be highly toxic to E. coli cells, were inserted into the expression vector pMAL-p2. The recombinant PAPs (rPAPMC and rPAPM) were successfully expressed in E. coli at 25 degrees C, being exported to the periplasm as soluble fusions with maltose binding protein (MBP). The rPAPs were cleaved from MBP by treatment with factor Xa and subsequently purified with final yields of 4.0 mg/liter (rPAPMC) and 5.5 mg/liter (rPAPM). rPAPM was resistant to protease digestion, but the C-terminal extrapeptide appeared to be susceptible and was partially digested by some protease in E. coli. Both rPAPMC and rPAPM were as active as the native PAPM from pokeweed leaves in depurinating rat liver and E. coli ribosomes, while the activities of rPAPMC on both ribosomes were decreased at least 60-fold by fusion with MBP. PMID- 10478457 TI - Natural antioxidant, chlorogenic acid, protects against DNA breakage caused by monochloramine. AB - Chlorogenic acid prevented a stepwise conversion of plasmid pUC18 DNA, from I- >form II-->form III, induced by 3 mM monochloramine with a half inhibition of 67.4 microM. Chlorogenic acid reacted with monochloramine in a time-dependent manner, and the reaction rate increased with decreasing pH. These results suggest that chlorogenic acid prevents genotoxicity of monochloramine in gastric mucosa. PMID- 10478458 TI - Effect of dietary taurine on endogenous hypercholesterolemia in rats fed on phenobarbital-containing diets. AB - The effect of dietary taurine on endogenous hypercholesterolemia induced by a phenobarbital-containing diet was investigated. Supplemented taurine did not affect the concentrations of serum cholesterol, but further potentiated the accumulation of hepatic cholesterol in the hypercholesterolemic state induced by phenobarbital. It is suggested that taurine might amplify the hepatic cholesterogenesis in phenobarbital-induced hypercholesterolemia. PMID- 10478459 TI - Substrate specificity switching of the flagellum-specific export apparatus during flagellar morphogenesis in Salmonella typhimurium. AB - During flagellar morphogenesis in Salmonella typhimurium, the flagellum-specific anti-sigma factor FlgM is exported out of the cells only after completion of hook assembly. In this study, we examined the export of the flagellar proteins, FlgD (hook capping protein), FlgE (hook protein), FlgK and FlgL (hook-filament junction proteins), FliD (filament capping protein), and FliC (flagellin), before and after completion of hook assembly. Like the FlgM protein, the FlgK, FlgL, FliD, and FliC proteins are exported efficiently only after completion of hook assembly. On the other hand, the FlgD and FlgE proteins are exported efficiently before, but poorly after, hook completion. These results indicate that the export properties are different between these two groups and that their export order exactly parallels the assembly order of the hook-filament structure. We propose that the substrate specificity switching occurs in the flagellum-specific export apparatus upon completion of hook assembly. PMID- 10478460 TI - Carbohydrate-dependent hemolytic activity of the conjugate composed of a C-type lectin, CEL-I, and an amphiphilic alpha-helical peptide, 4(3)-beta Ala2. AB - A lectin-cationic peptide conjugate, 4(3)-CEL-I, was prepared from an invertebrate C-type lectin, CEL-I, and an amphiphilic alpha-helical peptide, 4(3) beta Ala2 [Ac-(Leu-Ala-Arg-Leu)3-beta Ala2]. When 4(3)-CEL-I was incubated with rabbit erythrocytes, hemolysis was observed, especially at basic pH. Inhibition experiment using some carbohydrates suggested that hemolytic activity of 4(3)-CEL I was caused by the interaction between 4(3)-beta Ala2 portion in the conjugate and the lipid bilayer after binding to the carbohydrate chains on the cell surface by the lectin activity of CEL-I. PMID- 10478461 TI - Structural study of PLGA [copoly (DL-lactic/glycolic acid)], a biodegradable polymer for parenteral sustained release preparations, by tandem mass spectrometry. AB - A copoly (DL-lactic/glycolic acid) (PLGA), with a weight-average molecular weight of about 8400, has been characterized using fast atom bombardment (FAB)-tandem mass spectrometry in order to determine the sequence. Because of the large molecular size, PLGA was partially hydrolyzed and the terminal hydroxyl groups in the resulting oligomer mixture acetylated as the indicator. The FAB spectrum of this sample showed a complex ion signal pattern containing monomer to octamer. Diagnostic product ions containing useful information for sequence determination were observed in collision-induced dissociation-MS/MS and MS/MS/MS of these oligomer ions. The results of analysis for dimers through pentamers showed that they have random sequences of lactic and glycolic acid, suggesting that the whole structure of PLGA also has a random sequence. PMID- 10478462 TI - A novel class of inhibitors for human steroid 5 alpha-reductase: phenoxybenzoic acid derivatives. I. AB - In a search for novel nonsteroidal inhibitors of human prostatic 5 alpha reductase, we found a new series of phenoxybenzoic acid derivatives to be potent human prostatic 5 alpha-reductase inhibitors. Among them, 4-(biphenyl-4 yloxy)benzoic acid derivatives (2n, YM-31758), 2o and 2s showed more potent inhibitory activities than finasteride with IC50 values of 0.87, 0.67 and 0.56 nM, respectively. The optimized structures for the phenoxybenzoic acid derivatives 2d-2i were calculated by molecular modeling analysis, and the favorable distance between the carbon of the carboxyl group and the centroid of the phenyl group (benzene ring C) was found to be in the 9-11 A range. PMID- 10478463 TI - AS-924, a novel orally active bifunctional prodrug of ceftizoxime. Synthesis and relationship between physicochemical properties and oral absorption. AB - Ceftizoxime (CZX), a parenteral cephalosporin, has potent and broad antibacterial activity. To improve its oral absorption, we synthesized a series of monofunctional and bifunctional prodrugs of CZX. In rabbits, urinary recovery after oral administration of CZX was improved by esterification of the carboxyl group at the C-4 position with various lipophilic moieties (monofunctional prodrugs), and was further increased by introduction of a hydrophilic L-alanine to the amino group on the thiazole ring at the C-7 position (bifunctional prodrugs). Least-squares analysis showed good parabolic correlations between log P and urinary recovery for monofunctional and bifunctional prodrugs, respectively. AS-924, a bifunctional prodrug with a pivaloyloxymethyl and L alanyl moiety had the best balance of lipophilicity and water-solubility for oral absorption among the prodrugs synthesized. PMID- 10478464 TI - Amino acids and peptides. LIV. Application of 2-adamantyl derivatives as protecting groups to the synthesis of peptide fragments related to Sulfolobus solifataricus ribonuclease. I. AB - The 2-adamantyloxycarbonyl group was employed for the protection of the epsilon amino group of Lys and the hydroxyl group of Tyr, and the 2-adamantyl ester was employed for the protection of the beta-carboxyl group of Asp in order to construct eight peptide segments as building blocks for the preparation of peptide fragments related to the sequence of Sulfolobus solifataricus Ribonuclease. The usefulness of the above protecting groups developed in our laboratory was confirmed. PMID- 10478465 TI - Amino acids and peptides. LV. Application of 2-adamantyl derivatives as protecting groups to the synthesis of peptide fragments related to Sulfolobus solifataricus ribonuclease. II. AB - Segment condensations were performed to construct peptide fragments related to Sulfolobus solifataricus Ribonuclease. At each condensation step, the new protecting groups were stable. The protected peptide fragments were treated with a low-high HF procedure to give the desired peptide fragments. These peptide fragments were also prepared by the solid-phase method, and the obtained peptides were compared with those obtained by the solution method. The peptide fragments obtained by the solution method were identical with those obtained by the solid phase method on analytical HPLC, indicating that the new protecting groups could be easily removed by HF, and no racemization occurred during the synthesis of the protected peptides. PMID- 10478466 TI - Effects of incorporation of various amphiphiles into recipient liposome membranes on inter-membrane protein transfer. AB - To obtain information about the factors governing spontaneous inter-membrane protein transfer, we examined the effects of incorporation of various amphiphilic compounds in dimyristoylphosphatidylcholine (DMPC) liposomes on protein transfer from influenza virus-infected cells to the liposomes, and analyzed the physical properties of these liposome membranes. The incorporation of amphiphilic compounds, negatively charged dicetylphosphate (DCP), dipalmitoylphosphatidylserine (DPPS) or positively charged dimethyldipalmitoylammonium (DMDPA), into DMPC liposomal membranes enhanced protein transfer. The liposomes containing DCP, DPPS or DMDPA were unaffected by osmotic shock caused by external addition of glucose, suggesting a decrease in lipid packing in the liposomal membranes. Furthermore, calorimetric study of these liposomes showed that a phase separation occurred partially in the liposomal membranes. Accordingly, the membranes of DMPC liposomes containing DCP, DPPS and DMDPA should be distorted due to the coexistence of two phases, gel and liquid crystalline, in the membranes. Consequently, the membrane distortion could be responsible for the enhancement effects of the amphiphiles on the inter membrane protein transfer from influenza virus-infected cells to the liposomes. PMID- 10478467 TI - Phenolic constituents of Cassia seeds and antibacterial effect of some naphthalenes and anthraquinones on methicillin-resistant Staphylococcus aureus. AB - Thirteen phenolic glycosides including six new compounds were isolated from seeds of Cassia tora (Leguminosae). The structures of the new compounds, rubrofusarin triglucoside (7), nor-rubrofusarin gentiobioside (9), demethylflavasperone gentiobioside (10), torachrysone gentiobioside (11), torachrysone tetraglucoside (12) and torachrysone apioglucoside (13), were elucidated on the basis of spectroscopic and chemical evidence. The effects of the phenolic glycosides, their aglycones and several other compounds structurally related to them on Escherichia coli K12, Pseudomonas aeruginosa PAO1 and some strains of Staphylococcus aureus were then examined. Among them, torachrysone (15), toralactone (16), aloe-emodin (18), rhein (19) and emodin (20) showed noticeable antibacterial effects on four strains of methicillin-resistant Staphylococcus aureus with a minimum inhibitory concentration of 2-64 micrograms/ml. On the other hand, the phenolic compounds tested did not show strong antibacterial effects on E. coli and P. aeruginosa. PMID- 10478468 TI - Pregnanes and pregnane glycosides from Hoya carnosa. AB - Eleven pregnanes were isolated from the hydrolysate of the CHCl3 extract fractionated from the caules of Hoya carnosa. Among these, six pregnanes, including 19-acetoxydigipurpurogenin II, were new, and their structures were elucidated. The structures of twenty new pregnane tetraosides and pentaosides, named hoyacarnosides A-T, besides three known ones from the CHCl3 extract, were determined. PMID- 10478470 TI - Synthesis of optically active NC-1800, a therapeutic agent for urinary disturbance. AB - A new synthetic method for chiral oxazolidinone derivatives, therapeutic agents for treating urinary disturbance, is described. The condensed compound obtained from chiral 1-amino-3-phenyl-2-propanol and 1-phenyl-3-morpholino-1-propanone was reduced with Me4NBH(OAc)3 to give the intermediate, 1-(3-morpholino-1 phenylpropyl)amino-3-phenyl-2-propanol (MAPP) in 34% diastereomeric excess (d.e.). MAPP was converted to an urethane and purified by recrystallization of its methanesulfonate, to afford a single isomer, (2R)-1-[N-[(1S)-3-morpholino-1 phenylpropyl]-N -ethoxycarbonyl]amino-3-phenyl-2-propanol methanesulfonate (4-A. methanesulfonate). PMID- 10478469 TI - Amino acids and peptides. LIII. Synthesis and biological activities of some pseudo-peptide analogs of PKSI-527, a plasma kallikrein selective inhibitor: the importance of the peptide backbone. AB - Pseudo-peptide analogs of trans-4-aminomethylcyclohexanecarbonyl-L-phenylalanyl-4 aminopheny l acetic acid (PKSI-527, plasma kallikrein selective inhibitor), in which an amide bond (peptide bond) has been replaced by a CH2-NH bond, i.e., trans-4-aminomethylcyclohexanecarbonyl-L-phenylalanyl-psi (CH2-NH)-4-aminophenyl acetic acid (I), trans-4-aminomethylcyclohexanecarbonyl-psi (CH2-NH)-L phenylalanyl-4-aminophenyl acetic acid (II) and trans-4 aminomethylcyclohexanecarbonyl-D-phenylalanyl-psi (CH2-NH)-4-aminophenyl acetic acid (III) were synthesized. These pseudo-peptide analogs did not exhibit any detectable inhibitory activity against plasma kallikrein (PK), plasmin (PL), urokinase (UK), thrombine (TH) or trypsin (TRY). These results indicate that both carbonyl groups in the PKSI-527 are important for the manifestation of potent inhibitory activity against plasma kallikrein. PMID- 10478471 TI - Enzymatic synthesis of (-)- and (+)-acetoxyhexamides and (-)- and (+) hydroxyhexamides. AB - The enantioselective hydrolysis of (+/-)-4-(1-acetoxyethyl)-N (cyclohexylcarbamoyl)-benzenesulfona mides 3 with lipase Amano P from Pseudomonas sp. in a water-saturated solvent gave (R)-4-(1-hydroxyethyl)-N (cyclohexylcarbamoyl)benzenesulfonamide 2 (39%, > 99% ee) and unchanged (S)-3 (50%, 62% ee). On the other hand, enantioselective esterification of (+/-)-2 with lipase Amano P in the presence of vinyl acetate provided (R)-3 (41%, > 99% ee) and unchanged (S)-2 (46%, 78% ee). PMID- 10478472 TI - Glycerophosphocholines of the earthworm, Pheretima asiatica. AB - The lipid composition of the earthworm, Pheretima asiatica (Annelida), was analyzed. Four glycerophospholipids, together with four known glycosphingolipids, were isolated in pure form. The former four were 1-alkyl 2-acyl glycerophosphocholines possessing a C17:0 and/or C18:1 fatty acid residue. Their structures, including the position and geometry of the double bond, were determined on the bases of chemical and spectral data. PMID- 10478473 TI - HPLC analysis of juzen-taiho-to and its variant formulations and their antimetastatic efficacies. AB - Our previous study demonstrated that the oral administration of Juzen-taiho-to resulted in a significant inhibition of the liver metastasis of colon 26-L5 cells as compared with the untreated control, without side effects. We attempted to investigate the relationship between the HPLC pattern (referred to as the fingerprint) of the formulation and its component crude drugs and the inhibition of tumor metastasis in order to obtain the optimal efficacy and constant quality of the formulation. Two Juzen-taiho-to formulations (batches #1 and #2), which were individually prepared using the same 10 crude drugs and the same preparation procedure, showed similar anti-metastatic effects and absorbance patterns by HPLC analysis. Some variant formulations of Juzen-taiho-to, in which one crude drug was substituted with other crude drugs from different sources or places of origin, exhibited reduced efficacy as compared with the original formulation, as well as differences in the fingerprint pattern compared with the original formulation. Juzen (Naimo-Ogi-->Kibana-Ogi), a variant formulation with the substitution of Astragali radix of a different origin and place of harvest, showed significant inhibition of the liver metastasis of tumor cells and a HPLC fingerprint pattern similar to that of the original formulation. Thus, HPLC fingerprint analysis of Kampo medicines may provide a useful basis for obtaining their optimal efficacy as well as constant quality of the formulation, although it has some problems and limitations, such as detectability by and sensitivity to UV absorbance. PMID- 10478474 TI - Structural elucidation of the ceramide moiety of starfish gangliosides by collision-induced dissociation of the sodium ion complex. AB - Collision-induced dissociation (CID) spectra of sodium ion complexes ([M+Na]+ ions), produced by FAB-MS of methyl ester derivatives of ganglioside, indicate the length of the fatty acyl chain of the ceramide moieties without chemical degradation. In the case of a genuine ganglioside, only the fission of the glycosyl linkage of sialic acid was prominently observed. PMID- 10478475 TI - Synthesis of conformationally restricted analogs of baclofen, a potent GABAB receptor agonist, by the introduction of a cyclopropane ring. AB - Conformationally restricted analogs of baclofen (2), i.e., 5, 6, and their enantiomers ent-5, and ent-6, the conformations of which were restricted by introducing a cyclopropane ring, were designed as potential GABAB receptor ligands. Reaction of (R)-epichlorohydrin [(R)-7] and (4-chlorophenyl)acetonitrile in the presence of NaNH2 in benzene/tetrahydrofuran gave chiral cyclopropane derivatives 11 and 12, which were then converted into the target compounds 5 and 6, respectively. Their corresponding enantiomers, ent-5 and ent-6, were also synthesized starting from (S)-epichlorohydrin [(S)-7]. PMID- 10478476 TI - Synthesis of pyrazinone ring-containing opioid mimetics and examination of their opioid receptor-binding activity. AB - Cyclization of dipeptidyl chloromethyl ketones gave 6-(4-aminobutyl)-3 carboxyethyl-5-methyl-2(1H)-pyrazinone, 3-(4-aminobutyl)-6-carboxyethyl-5-methyl 2(1H)-pyrazinone, and 3,6-bis(4-aminobutyl)-5-methyl-2(1H)-pyrazinone, which were inserted into the enkephalin sequence to give opioid mimetics. Thus, it was confirmed that a pyrazinone ring can be easily inserted into a peptide sequence in order to evaluate structural components required for biologically active peptides. PMID- 10478477 TI - dnaK and the heat stress response of Pseudomonas syringae pv. glycinea. AB - The dnaK gene from Pseudomonas syringae pv. glycinea PG4180 was cloned and sequenced. The dnaK coding region was 1,917 bp and contained a putative sigma 32 heat shock promoter 86 bp upstream of the translational start site. grpE, another heat shock gene, was found immediately upstream of the putative dnaK promoter. The predicted amino acid sequence of dnaK showed relatedness to the ATPase and substrate binding domains commonly found in heat shock proteins, as well as the highly conserved signature sequence motifs belonging to the Hsp70 protein family. Furthermore, the PG4180 dnaK gene complemented an Escherichia coli dnaK mutant for growth at temperatures above 37 degrees C, indicating that a fully functional dnaK homologue had been cloned from P. syringae pv. glycinea. All attempts to eliminate dnaK function by insertion mutagenesis failed, possibly because DnaK performs essential functions in P. syringae pv. glycinea. Expression of dnaK in P. syringae pv. glycinea PG4180 was investigated by constructing dnaK::uidA transcriptional fusions; expression of dnaK increased markedly when cells were preincubated at 18 degrees C and then shifted to 35 degrees C. An anti-DnaK monoclonal antibody was used to detect DnaK; in P. syringae pv. glycinea race 4, DnaK levels followed cell density during a 6-h incubation at 26 degrees C. When cells were shifted from 26 degrees C to either 32 or 38 degrees C, DnaK levels increased transiently, and then decreased rapidly. Although the cells continued to grow when incubated at 32 degrees C, growth was not supported at 38 degrees C. Our results indicate that P. syringae pv. glycinea responds to heat shock by producing DnaK, but DnaK does not aid in acclimation to sustained elevated temperatures. PMID- 10478478 TI - The response regulator expM is essential for the virulence of Erwinia carotovora subsp. carotovora and acts negatively on the sigma factor RpoS (sigma s). AB - The main virulence factors of Erwinia carotovora subsp. carotovora, the secreted, extracellular cell-wall-degrading enzymes, are controlled by several regulatory mechanisms. We have isolated transposon mutants with reduced virulence on tobacco. One of these mutants, with a mutation in a gene designated expM, was characterized in this study. This mutant produces slightly reduced amounts of extracellular enzymes in vitro and the secretion of the enzymes is also affected. The expM wild-type allele was cloned together with an upstream gene, designated expL, that has an unknown function. The expM gene was sequenced and found to encode a protein with similarity to the RssB/SprE protein of Escherichia coli and the MviA protein of Salmonella typhimurium. These proteins belong to a new type of two-component response regulators that negatively regulate the stability of the Sigma factor RpoS (sigma s) at the protein level. The results of this study suggest that ExpM has a similar function in E. carotovora subsp. carotovora. We also provide evidence that the overproduction of RpoS in the expM mutant is an important factor for the reduced virulence phenotype and that it partly causes the observed phenotype seen in vitro. However, an expM/rpoS double mutant is still affected in secretion of extracellular enzymes, suggesting that ExpM in addition to RpoS also acts on other targets. PMID- 10478479 TI - Isolation of a cDNA encoding a beta-1,4-endoglucanase in the root-knot nematode Meloidogyne incognita and expression analysis during plant parasitism. AB - A beta-1,4-endoglucanase encoding cDNA (EGases, E.C. 3.2.1.4), named Mi-eng-1, was cloned from Meloidogyne incognita second-stage juveniles (J2). The deduced amino acid sequence contains a catalytic domain and a cellulose-binding domain separated by a linker. In M. incognita, the gene is transcribed in the migratory J2, in males, and in the sedentary adult females. In pre-parasitic J2, endoglucanase transcripts are located in the cytoplasm of the subventral esophageal glands. The presence of beta-1,4-endoglucanase transcripts in adult females could be related to the expression of the gene in esophageal glands at this stage. However, cellulase activity within the egg matrix of adult females suggests that the endoglucanase may also be synthesized in the rectal glands and involved in the extrusion of the eggs onto the root surface. The maximum identity of the predicted MI-ENG-1 catalytic domain with the recently cloned cyst nematode beta-1,4-endoglucanases is 52.5%. In contrast to cyst nematodes, M. incognita pre parasitic J2 were not found to express a beta-1,4-endoglucanase devoid of a cellulose-binding domain. PMID- 10478481 TI - An hrcU-homologous gene mutant of Xanthomonas campestris pv. glycines 8ra that lost pathogenicity on the host plant but was able to elicit the hypersensitive response on nonhosts. AB - Transposon mutagenesis was used to isolate nonpathogenic mutants of Xanthomonas campestris pv. glycines 8ra, which causes bacterial pustule disease in soybean. A 6.1-kb DNA region in which a mutation gave loss of pathogenicity was isolated and found to carry six open reading frames (ORFs). Four ORFs had homology with hrcU, hrcV, hrcR, and hrcS genes of Ralstonia solanacearum and X. campestris pv. vesicatoria. One nonpathogenic mutant, X. campestris pv. glycines H80, lost pathogenicity on soybean but was able to elicit the hypersensitive response (HR) on nonhost pepper and tomato plants. This mutant still multiplied as well as the wild type in the leaves or cotyledons of soybean. Although the DNA and amino acid sequences showed high homology with known hrp genes, the hrcU-homolog ORF is not required for HR induction on nonhost plants, pepper and tomato, or for the multiplication of bacteria in the host plant. This gene was only required for the pathogenic symptoms of X. campestris pv. glycines 8ra on soybean. PMID- 10478482 TI - Oligogalacturonide-mediated induction of a gene involved in jasmonic acid synthesis in response to the cell-wall-degrading enzymes of the plant pathogen Erwinia carotovora. AB - Identification of Arabidopsis thaliana genes responsive to plant cell-wall degrading enzymes of Erwinia carotovora subsp. carotovora led to the isolation of a cDNA clone with high sequence homology to the gene for allene oxide synthase, an enzyme involved in the biosynthesis of jasmonates. Expression of the corresponding gene was induced by the extracellular enzymes from this pathogen as well as by treatment with methyl jasmonate and short oligogalacturonides (OGAs). This suggests that OGAs are involved in the induction of the jasmonate pathway during plant defense response to E. carotovora subsp. carotovora attack. PMID- 10478483 TI - Synthesis and triplex forming properties of pyrimidine derivative containing extended functionality. AB - Two pyrimidine nucleosides have been synthesized containing extended hydrogen bonding functionality. In one case the side chain is based upon semicarbazide and in the second monoacetylated carbohydrazide was employed. DNA sequences could be prepared using both analogue nucleosides in a reverse coupling protocol, and provided that the normal capping step was eliminated and that the iodine-based oxidizing solution was replaced with one based upon 10-camphorsulfonyl oxaziridine. Both derivatives exhibited moderate effects in targeting selectively C-G base pairs embedded within a polypurine target sequence. PMID- 10478484 TI - Increased cytotoxicity and decreased in vivo toxicity of FdUMP[10] relative to 5 FU. AB - The efficacy of treatment with 5-Fluorouracil (5-FU) is limited, in part, by its inefficient conversion to 5-Fluoro-2'-deoxyuridine-5'-O-monophosphate (FdUMP). We present data indicating that FdUMP[10], designed as a pro-drug for intracellular release of FdUMP, is cytotoxic as a consequence of uptake of the multimeric form. FdUMP[10] is stable in cell culture medium, with more than one-half of the material persisting as multimers of at least six nucleotides after a 48 h incubation at 37 degrees C. FdUMP[10] is more than 400 times more cytotoxic than 5-FU towards human colorectal tumor cells (H630). FdUMP[10] also has decreased toxicity in vivo, with doses as high as 200 mg/kg/day (qdx3) administered to Balb/c mice without morbidity, compared to a maximum tolerated dose of 45 mg/kg/day for 5-FU using the same protocol. FdUMP[10] shows reduced sensitivity to OPRTase- and TK-mediated drug resistance, relative to 5-FU and FdU, respectively, and is much more cytotoxic than 5-FU towards cells that overexpress thymidylate synthase. Thus, FdUMP[10] is less susceptible to resistance mechanisms that limit the clinical utility of 5-FU. The increased cytotoxicity, decreased toxicity in vivo, and reduced sensitivity to drug resistance of FdUMP[10], relative to 5-FU, indicates multimeric FdUMP is potentially valuable as an anti-neoplastic agent, either as a single agent, or in combination with 5 FU. PMID- 10478485 TI - A phosphate bound universal linker for DNA synthesis. AB - A uridine-based linker immobilized onto polystyrene beads at the 5' terminus via a phosphodiester group and then used as a universal DNA synthesis support gives post synthesis DNA cleavage in 8 hrs or less without alkali metal salts. DNA produced with the new support was analyzed by HPLC, MALDI mass spectroscopy and PAGE. Each analysis showed DNA of equivalent quality to that produced with standard CPG supports, without contaminating materials resulting from linker or support backbone decomposition. PMID- 10478486 TI - Synthesis and biophysical studies of modified oligonucleotides containing acyclic amino alcohol nucleoside analogs. AB - Novel serine derivative of thymine was prepared and incorporated into oligonucleotides. These modified oligonucleotides were studied for their binding affinity with complementary DNA/RNA. PMID- 10478487 TI - Substrate/inhibitor properties of human deoxycytidine kinase (dCK) and thymidine kinases (TK1 and TK2) towards the sugar moiety of nucleosides, including O'-alkyl analogues. AB - Nucleoside analogues with modified sugar moieties have been examined for their substrate/inhibitor specificities towards highly purified deoxycytidine kinase (dCK) and thymidine kinases (tetrameric high-affinity form of TK1, and TK2) from human leukemic spleen. In particular, the analogues included the mono- and di-O' methyl derivatives of dC, dU and dA, syntheses of which are described. In general, purine nucleosides with modified sugar rings were feebler substrates than the corresponding cytosine analogues. Sugar-modified analogues of dU were also relatively poor substrates of TK1 and TK2, but were reasonably good inhibitors, with generally lower Ki values vs TK2 than TK1. An excellent discriminator between TK1 and TK2 was 3'-hexanoylamino-2',3'-dideoxythymidine, with a Ki of approximately 600 microM for TK1 and approximately 0.1 microM for TK2. 3'-OMe-dC was a superior inhibitor of dCK to its 5'-O-methyl congener, consistent with possible participation of the oxygen of the (3')-OH or (3')-OMe as proton acceptor in hydrogen bonding with the enzyme. Surprisingly alpha-dT was a good substrate of both TK1 and TK2, with Ki values of 120 and 30 microM for TK1 and TK2, respectively; and a 3'-branched alpha-L-deoxycytidine analogue proved to be as good a substrate as its alpha-D-counterpart. Several 5'-substituted analogues of dC were good non-substrate inhibitors of dCK and, to a lesser extent, of TK2. Finally, some ribonucleosides are substrates of the foregoing enzymes; in particular C is a good substrate of dCK, and 2'-OMe-C is an even better substrate than dC. PMID- 10478489 TI - Lactotroph hyperplasia in the pituitaries of female mice expressing high levels of bovine growth hormone. AB - PEPCK/bGH transgenic mice have very high blood levels of foreign GH, and prominent reproductive disturbances, especially in females. To obtain a deeper insight into the causes of these abnormalities, pituitaries of PEPCK/bGH transgenics were studied by immunocytochemistry, electron microscopy and in situ hybridization (ISH) techniques. Pituitary weights were significantly reduced (P < 0.05) in transgenic males, while in transgenic females they were increased without reaching significance compared to nontransgenic controls. In both sexes, GH cells were inhibited, as previously described in other lines of GH transgenic mice. In females, PRL cells were increased by 37% compared to controls. Ultrastructurally, the lactotrophs had characteristics of stimulation and PRL mRNA was increased by 35%. In males the increase in the number of PRL immunoreactive cells was not significant, the PRL mRNA signal did not differ from controls, and there were no changes in their ultrastructure. Only in females ACTH cells were significantly reduced (P < 0.05) in number and unchanged in males; however, POMC mRNA signal was increased in both genders and reached significance (P < 0.05) in males. In females, but not in males, the percentage of LH cells was lower than in control mice. In conclusion, the high blood bGH levels induced sex related changes in transgenic mice from the present line. The infertility of PEPCK/bGH transgenic females may be attributed to lactotroph hyperplasia and marked reduction in number of gonadotrophs. PMID- 10478490 TI - Turnip mosaic potyvirus resistance in Nicotiana benthamiana derived by post transcriptional gene silencing. AB - The coat protein (CP) gene of turnip mosaic potyvirus isolate ESC8 (TuMV-ESC8) was cloned and sequenced. Comparisons of the 867-nucleotide (nt) CP region with those of 11 TuMV isolates showed 86.7-89.3% nucleotide identity and 92.4-95.5% amino acid identity. The CP gene was cloned into a plant expression vector and transformed into Nicotiana benthamiana plants via Agrobacterium tumefaciens mediated leaf disk transformation. Progeny from R0 lines was screened for resistance to TuMV-ESC8. Five of 29 tested lines showed TuMV protection in more than 50% of their progeny. Interestingly, some of the resistant plants transformed with the CP gene of TuMV displayed mild mosaicism in the new growing leaves at the later stages of evaluation; but these mosaic symptoms disappeared when the leaves were fully expanded. Collective data from steady-state RNA analysis and nuclear run-on assay of a line showed that the resistance was RNA mediated through the post-transcriptional gene silencing mechanism. PMID- 10478491 TI - Enrichment and efficient screening of ES cells containing a targeted mutation: the use of DT-A gene with the polyadenylation signal as a negative selection maker. AB - Gene targeting in embryonic stem (ES) cells via homologous recombination can occur at very low frequency. In order to enrich homologous recombinants before screening, a negative selection marker, such as thymidine kinase (TK) and diphtheria toxin A fragment (DT-A), has been commonly used. In this study, we developed a negative selection marker using DT-A gene with polyadenylation signal and it was designated DT-ApA. To determine the difference in targeting efficiency of the negative selections, we constructed three different targeting vectors for each negative selection (first, TK at the 3' end, second, TK at both the 5' and 3' ends < 2 X TK >, and third, DT-ApA at the 5' end of the homologous sequences). Gene targeting experiments using these constructs clearly showed that negative selection using DT-ApA was more efficient than that using TK for homologous recombination and that negative selection using DT-ApA was as efficient as that using 2 X TK. Considering the fact that the use of DT-ApA is more convenient for construction of targeting vectors than that of 2 X TK, DT-ApA is an efficient negative selection marker. In addition, we examined long and accurate PCR (LA PCR) for screening gene targeted clones. The use of LA-PCR with genomic DNAs from ES cell clones facilitated simple detection of homologous recombinants, suggesting that the screening with LA-PCR is compatible with the use of longer homologous sequences of both arms in vector design. Our results indicate that the use of DT-ApA for negative selection together with the application of LA-PCR for screening ensures efficient and time-saving screening for homologous recombinants. PMID- 10478488 TI - Internal ribosome entry sites (IRESs): reality and use. AB - IRESs are known to recruit ribosomes directly, without a previous scanning of untranslated region of mRNA by the ribosomes. IRESs have been found in a number of viral and cellular mRNAs. Experimentally, IRESs are commonly used to direct the expression of the second cistrons of bicistronic mRNAs. The mechanism of action of IRESs is not fully understood and a certain number of laboratories were not successful in using them in a reliable manner. Three observations done in our laboratory suggested that IRESs might not work as functionally as it was generally believed. Stem loops added before IRESs inhibited mRNA translation. When added into bicistronic mRNAs, IRESs initiated translation of the second cistrons efficiently only when the intercistronic region contained about 80 nucleotides, and they did not work any more effectively with intercistronic regions containing at least 300-400 nucleotides. Conversely, IRESs inserted at any position into the coding region of a cistron interrupted its translation and initiated translation of the following cistron. The first two data are hardly compatible with the idea that IRESs are able to recruit ribosomes without using the classical scanning mechanism. IRESs are highly structured and cannot be scanned by the 40S ribosomal subunit. We suggest that IRESs are short-circuited and are essentially potent stimulators favoring translation in particular physiological situations. PMID- 10478492 TI - Association of the 5'HS4 sequence of the chicken beta-globin locus control region with human EF1 alpha gene promoter induces ubiquitous and high expression of human CD55 and CD59 cDNAs in transgenic rabbits. AB - Whatever its field of application, animal transgenesis aims at a high level of reproducible and stable transgene expression. In the case of xenotransplantation, prevention of hyperacute rejection of grafts of animal origin requires the use of organs expressing human inhibitors of complement activation such as CD55 (DAF) and CD59. Pigs transgenic for these molecules have been produced, but with low and variable levels of expression. In order to improve cDNA expression, a vector containing the 5'HS4 region from the LCR of the chicken beta-globin locus and the promoter and the first intron from the human EF1 alpha gene, was used to co express human CD55 and CD59 cDNAs in transgenic rabbits. The transgenic lines with the 5'HS4 region displayed dramatically enhanced CD55 and CD59 mRNA concentrations in brain, heart, kidney, liver, lung, muscle, spleen and aortic endothelial cells in comparison with the transgenic lines without the 5'HS4 region. In the absence of the 5'HS4 region, only some of the transgenic lines displayed specific mRNAs and at low levels. Human CD55 and CD59 proteins were detectable in mononuclear cells from transgenic rabbits although at a lower level than in human mononuclear cells. On the other hand, primary aortic endothelial cells from a bi-transgenic line were very efficiently protected in vitro against human complement-dependent lysis. Transgenic rabbits harbouring the two human inhibitors of complement activation, CD55 and CD59, can therefore be used as new models in xenotransplantation. Moreover, the vector containing the 5'HS4 region from the LCR of the chicken beta-globin locus seems appropriate not only for xenotransplantation but also for any other studies involving transgenic animals in which cDNAs have to be expressed at a high level in all cell types. PMID- 10478493 TI - Expression of human blood clotting factor VIII in the mammary gland of transgenic sheep. AB - By targeting the expression of sequences encoding non-milk proteins to the mammary gland of transgenic farm animals, the organ could serve as a 'bioreactor' for producing pharmacologically active proteins on a large scale. Here we report the generation of transgenic sheep bearing a fusion gene construct with the human blood clotting factor VIII (hFVIII) cDNA under the transcriptional control of a 2.2 kb fragment of the mammary gland specific promoter of the ovine beta Lactoglobulin (beta-Lac) gene. Six founder animals were generated bearing a hFVIII cDNA construct with the introns of the murine metallothionein (MtI) gene (beta-Lac/hFVIII-MtI). Founders transmitted the transgene in a Mendelian fashion and two transgenic lines were generated. Ten out of 12 transgenic F1-females expressed rhFVIII mRNA in exfoliated mammary epithelial cells isolated from the milk. But only in transgenic F1 ewes 4010 and 603 hFVIII clotting activity estimated at 4-6 ng/ml was detected in defatted milk. Furthermore, the presence of rhFVIII-protein in ovine milk was demonstrated by a specific band at approximately 190 kD following immunoprecipitation and immunoblotting. Transgenic founder 395 expressed rhFVIII mRNA in biopsied mammary gland tissue, in exfoliated mammary cells as well as ectopically in brain, heart, spleen, kidney and salivary gland, suggesting that the employed beta-Lac promoter fragment lacks essential sequences for directing expression exclusively to the mammary gland. A rhFVIII standard preparation (rhFVIIIstd) was rapidly sequestered in a saturable fashion in ovine milk, thus rendering it largely inaccessible to immunoprecipitation although its biological activity was retained. Recovery of hFVIIIstd was dependent on milk donor, storage temperature and dilution of milk sample. PMID- 10478494 TI - The emotional impact of sexual violence research on participants. AB - Participation in research on sexual trauma may provoke disturbing memories and distressing emotions. Despite the proliferation of research on sexual violence during the last decade, little is known about the effects of study involvement on participants. Based on a review of the literature, the author's experiences as a sexual as a violence, researcher, and reflections of women who have participated in such research, this article explores the emotional impact of sexual violence research on participants. Although the risk of lasting harm stemming from participation in trauma research is a legitimate concern, the benefits of confiding a traumatic experience to a trustworthy other seem to outweigh the immediate distress that accompanies discussion of painful experiences. A useful framework for understanding the responses of research participants who talk or write about traumatic experiences is discussed. PMID- 10478495 TI - Relationships between aggressive behavior in cognitively impaired nursing home residents and use of restraints, psychoactive drugs, and secured units. AB - This study examined relationships between aggressive behavior in cognitively impaired nursing home residents and physical restraints, psychoactive drugs, and placement on a secured unit. Data were obtained from 116 residents who were consistently aggressive as measured by the Ryden Aggression Scale 2. Subjects averaged 9.5 aggressive behaviors per day. Forty-seven percent of subjects were restrained, and 62% were regularly receiving psychoactive drugs. Use of restraints, antipsychotics, and placement on a secured unit were all significantly related to increased physical aggression scores. Four variables accounted for 23% of the variance in physical aggression scores: location on a secured unit, not receiving an antidepressant, being restrained, and number of psychotropic and/or anxiolytic medications administered. Significantly lower physical aggression scores were noted for subjects receiving antidepressants. PMID- 10478496 TI - The development and clinical testing of an outpatient program for women with bulimia nervosa. AB - This article describes the development of a cost-effective treatment program for women with Bulimia Nervosa (BN). A comprehensive review of the literature was undertaken to determine the most effective programs. The Time Limited Cognitive Behavioral Program (TLCBP) was developed and clinically tested on two groups of women to determine its cost-effectiveness. The program consisted of a 12-week intervention phase followed by a 14-week support/reinforcement phase. The clinical findings were that 9 of the 10 clients were abstinent following the 12 week intervention. Other symptoms improved following the 14-week phase of support/reinforcement. Utilizing an Advanced Practice Psychiatric-Mental Health Registered Nurse (APRN) significantly reduced the cost of the treatment, as did the use of a group model approach. APRN's have bio-behavioral knowledge as well as research skills. They are in a unique position to conduct outcome research with women with BN, as well as with clients who have other disorders with both biological and behavioral symptoms. PMID- 10478497 TI - Testing a model of family stress and coping based on war and non-war stressors, family resources and coping among Lebanese families. AB - This study was undertaken to describe the objective stressors, perceived stress, coping, and resources of families living in Beirut during the Lebanese war (1975 1991) and to test a model predicting the relationships of these variables to family adaptation. The sample consisted of 438 families chosen at random. Independent variables included objective stressors and perceived stress. The mediating variables were family resources and coping strategies. The dependent variables were health and interactional indicators of family adaptation: physical and psychological health, depression, and interpersonal and marital relationships. Findings provided support for the theoretical framework. Multiple regression analyses revealed that perceived stress, rather than the objective occurrence of events, predicted family adaptation. Family resources, particularly social support, positively impacted family adaptation and was associated with increased use of cognitive coping. The findings provide a theoretical model which, on further testing, can serve as a basis for practice by health professionals when working with traumatized families. PMID- 10478498 TI - The suitability of United States Pharmacopoeia Dispensory Information psychotropic drug leaflets for urban patients with limited reading skills. AB - Although considerable research exist on the readability of patient education materials, few studies have focused on the efficacy of information drug leaflets used for psychopharmacology education of patients with limited reading skills. The purpose of this investigation was to evaluate the suitability of United States Pharmacopoeia Dispensary Information (USP-DI) drug leaflets (N = 42) for educating urban in-patients at a psychiatric unit using measurement criteria of the Suitability Assessment for Materials (SAM) instrument. The findings showed the USP-DI information drug leaflets were unsuitable as an educational tool for psychiatric in-patients with poor reading ability. PMID- 10478499 TI - Self-resolution of alcohol problems as a process of investing and re-investing in self. AB - Resolution of alcohol problems without formal treatment or participation in support groups was studied using a grounded theory approach. Thirteen former problem drinkers and their close associates were interviewed, and verbatim transcripts were analyzed using grounded theory methods. The core of self resolving alcohol problems involves investing and/or reinvesting in self to gain or regain the essence of who or what one wants to be. Self-resolvers tend to independently change for their own benefit rather than the benefit of others, and personal resources may promote or delay the process. The effort required to self resolve drinking problems varies, and close associates tend to be minority investors in the process. Input from health-care providers regarding the health related hazards of drinking are largely ignored because threats to who or what one wants to be are perceived as more significant. As such, clinical interventions should emphasize personally relevant threats to self versus focusing solely on the physical consequences of alcohol abuse. PMID- 10478500 TI - [Quality of life of psychiatric patients]. PMID- 10478502 TI - Pan American Conference on Antimicrobial Resistance in the Americas. PMID- 10478501 TI - Leading causes of mortality on the United States--Mexico border. PMID- 10478503 TI - Blood safety in the Americas. PMID- 10478504 TI - News: partnership for measles eradication in the Americas by the year 2000 PAHO and the CDC of the United States. PMID- 10478505 TI - Norms and standards in epidemiology: guidelines for epidemiological surveillance. PAHO Communicable Diseases Program. PMID- 10478507 TI - Preparatory meeting of the PAHO/WHO Regional Advisory Committee on Health Statistics (CRAES). PMID- 10478508 TI - An academic-community partnership: a model of service and education. AB - To meet the challenge of preparing nurses for delivery of health care that is directed toward health promotion and focused on populations at the community level, it is critical that academicians develop new methods to educate their students. In this article, I describe an innovative clinical practice model in which an academic-community partnership was created between a college of nursing and a neighborhood grade school and parish. The purpose of the partnership is to provide needed health services to clients, at the same time giving students the opportunity to practice population-focused care in the community. The benefits of the partnership are numerous, including improved client health status, increased access to health promotion services, and enhanced student learning. PMID- 10478509 TI - A comparison of HIV/AIDS knowledge among high school freshmen and senior students. AB - Statistical data show that heterosexual transmission of AIDS among teenagers is a significant problem. The purpose of this study was to assess and compare the level of knowledge concerning HIV/AIDS among high school freshmen- and senior level students and to determine the association between certain demographic variables and the students' knowledge level. A convenience sample of 169 freshmen and 274 senior high school students were surveyed at a local area high school (N = 443). A 49-item knowledge questionnaire, used by the high school district HIV/Abstinence program, and a demographic questionnaire developed by the investigators were used to collect the data. The findings revealed that both the freshmen and senior students had several misconceptions about HIV/AIDS, including the modes of transmission, the sure way of preventing the sexual transmission, donating blood, and the usual causes of death for people with AIDS. A comparison of the mean knowledge scores among freshmen and senior students indicated there was no significant difference among the two groups. The mean knowledge score of the seniors was significantly affected by whether they had received HIV/AIDS education. However, this was not the case for the freshmen. African American students, especially the female students, had significantly lower knowledge scores than other ethnic groups. PMID- 10478510 TI - Solution-focused approaches to tobacco reduction with disadvantaged prenatal clients. AB - Despite high rates of smoking among socioeconomically disadvantaged pregnant women, few tobacco cessation programs have been geared for this client group. This article describes newly developed strategies for addressing tobacco reduction with disadvantaged, high-risk prenatal clients enrolled in a community based Pregnancy Outreach Program in a northern Canadian city. The new tobacco reduction strategies were based on "solution-focused" approaches to interviewing and counseling. Process evaluations conducted with program staff revealed that solution-focused approaches provided effective, client-centered, respectful ways of discussing tobacco reduction with disadvantaged clients. Solution-focused approaches also positively impacted clients' sense of self-efficacy regarding tobacco reduction. The benefits of using solution-focused approaches as an alternative approach to tobacco reduction with disadvantaged prenatal clients are discussed. PMID- 10478511 TI - Motivating the well elderly to exercise. AB - Most older adults hope to remain independent until death, but their inactivity, poor nutrition, and disabling diseases contribute to the loss of functions that are necessary for independence. Although exercise has been shown to improve a person's functioning and decrease health care costs, two thirds of persons over age 65 do not exercise regularly. The purpose of this article is to describe intervention strategies and evaluate their usefulness in motivating adherence to an exercise program designed specifically for well-elderly community dwellers. Attendance records were reviewed and informal discussions were held to determine the usefulness of the selected intervention strategies. Fifty-seven percent of the group's members attended at least one half of the 52 sessions indicating that our strategies were effective. Information about the exercise strategies that we used can help community health providers to promote health by designing exercise programs that will entice the elderly to attend on a regular basis. PMID- 10478512 TI - Parkinson's disease and the U.S. health care system. AB - Parkinson's disease is an expanding healthcare problem in our society. With the burgeoning elderly population in this country, it is important for community health nurses to understand the management of these patients. Although the cause of idiopathic Parkinson's disease is still unknown, there is much new research developing in this area. Local and national foundations are available to assist the patient and caregiver in education, referrals, and support groups. The effects of this disease process on the patient and family are profound. New government legislation may offer a promising link to researchers and practitioners nationwide. Healthcare implications are varied, and nurses need to keep abreast of changes in Medicare and managed care systems to ensure that homebound clients receive needed services. Community health nurses provide a vital link in the healthcare system for the Parkinson's patient. PMID- 10478513 TI - Leukotriene modifiers in the management of asthma. AB - Leukotrienes play a major role in the underlying pathophysiology of asthma. Leukotriene-receptor antagonists and direct leukotriene-synthesis inhibitors may be important new agents in maintenance therapy for asthma. Their role may also extend to attenuating exercise-induced and allergen-induced bronchospasm. This article reviews the effectiveness of available leukotriene modifiers in various clinical studies, reviews their adverse effects profile, and discusses their potential roles in the management of asthma. PMID- 10478514 TI - Efficacy and safety of azelastine nasal spray for the treatment of allergic rhinitis. AB - Azelastine hydrochloride is a nasally administered antihistamine that is effective and safe for the treatment of perennial and seasonal allergic rhinitis. In addition to acting as a histamine H1-receptor antagonist, azelastine also inhibits the production or release of many chemical mediators of the allergic response such as leukotrienes, free radicals, and cytokines. After nasal administration, azelastine is systemically absorbed with a bioavailability of about 40%. The side effects of azelastine are drowsiness, headache, and bitter taste. Azelastine has a rapid onset of action with a benefit in about 2 hours and a prolonged duration of activity (12 to 24 hours). Studies have shown azelastine to be more effective than placebo in terms of reduction of the major and total symptom complexes of allergic rhinitis. Comparison studies have demonstrated that azelastine is as effective as ebastine, loratadine, cetirizine hydrochloride, and terfenadine at symptom reduction, with varying results when compared with the corticosteroids budesonide and beclomethasone. Although there are conflicting studies, some have demonstrated that azelastine reduces the nasal congestion of allergic rhinitis. This feature that distinguishes it from oral antihistamines is of great interest because corticosteroids are known to be quite effective for the relief of nasal congestion, whereas the antihistamines are effective for the sneezing, itchy eyes, itchy nose, and watery eyes, but not the congestion. Azelastine nasal spray seems to be an efficacious treatment for allergic rhinitis with a rapid onset and long duration of activity, but without the systemic adverse effects of traditional sedating antihistamines. PMID- 10478515 TI - Seasonal allergic conjunctivitis: overview and treatment update. AB - Seasonal allergic conjunctivitis is rarely associated with permanent vision impairment; however, it is a relatively common condition that may compromise the quality of life. It may, in extreme cases, impair daily activities, including work. Numerous treatment options have become available for the relief of acute symptoms. Avoidance should always be the first line in therapy but, in most cases, is not practical, especially with pollen allergies. The use of saline eyedrops is simple and nontoxic, and it is effective in up to 30% to 35% of cases. It can and should be added to all other remedies to reduce adverse effects and cost by decreasing the need for other therapeutic options. Antihistamines and decongestants are useful treatment choices for the majority of cases. Ketorolac tromethamine may be helpful in relieving pruritus, but it offers little advantage over topical antihistamines. Corticosteroids may be used for severe cases for a limited time. If topical corticosteroids are being considered, an ophthalmologist should be consulted. Cromolyn sodium and lodoxamide ophthalmic solution may be helpful in the prophylaxis of symptoms during the allergy season, but these agents require frequent dosing. Olopatadine hydrochloride is a mast cell stabilizer and antihistamine that can be dosed twice a day. Immunotherapy is effective and should be offered to those who are intolerant or have allergic conjunctivitis refractory to medications. PMID- 10478516 TI - Behcet's disease and pheochromocytoma. AB - A 46-year-old black female with a history of Behcet's Disease was admitted with a diagnosis of abdominal pain. She had complained of palpitations, diaphoresis, anxiety, and fatigue for years which had been variously attributed to the perimenopause and her Behcet's. During her hospital evaluation, she had an episode of supraventricular tachycardia associated with severely elevated blood pressure. Subsequent evaluation revealed a large pararenal mass which was found to be a pheochromocytoma. Search of six databases revealed this to be only the second case report in the English literature of a pheochromocytoma in association with Behcet's Disease. PMID- 10478517 TI - Human monocytic ehrlichiosis in a thirteen year old: a case report. PMID- 10478518 TI - Infection with Chlamydia pneumoniae and atherosclerosis: a review. AB - There are numerous studies that were able to find C. pneumoniae infections as a contributing factor in atherosclerosis pathogenesis. Positive serology for C. pneumoniae was found in most studies in patients with atherosclerosis (IgG, IgA, IgM). Most studies were able to correlate an elevated IgA antibody titer rather than the IgG titer to the risk for atherosclerosis. C. pneumoniae and its components (DNA, antigens) were detected in atherosclerotic plaques using immunohistochemistry, PCR, electron microscopy and cell cultures. C. pneumoniae has been located in endothelium, smooth muscle cells and macrophages of arterial wall with atherosclerosis but not in normal arteries. Cellular models have shown that C. pneumoniae is able to replicate in endothelium, macrophages and smooth muscle cells. A high C. pneumoniae antibody titer was found to correlate with high level of LDL and triglycerides and low level of HDL. C. pneumoniae infection increases platelet adhesion and adhesion molecules at the surface of endothelium. C. pneumoniae could be a cofactor for atherosclerosis combined with high level of lipids as shown in an animal model but not alone. Strong cellular and humoral immunity have been found in men with atherosclerosis and positive C. pneumoniae titers. This organism could be found in diverse arteries with atherosclerosis. One particular C. pneumoniae strain (AR 39) appears to be more frequently involved in atherosclerosis. Antibiotic treatment with azithromycin appears to be protective against atherosclerosis complications. However, there is as yet no conclusive evidence that C. pneumoniae causes atherosclerosis, but most likely it may enhance this process. Two large randomized clinical trials are currently underway evaluating azithromycin treatment in patients with atherosclerosis which will hopefully give us answers about the role of antibiotic treatment. PMID- 10478519 TI - Does infection cause atherosclerosis--and most other diseases? PMID- 10478522 TI - [Leiomyomas of small bowel]. AB - The analysis of our own country and foreign literature, devoted to the problems of clinical aspects, diagnosis and treatment of leiomyomas of the small bowel for the last 48 years has been carried out. The authors present their personal experience in follow-up of 6 cases of the small bowel leiomyomas, the main clinical manifestations of which being recurrent intestinal bleedings and in lesser degree--small bowel obstruction. The results of preoperative examination evidence that besides thorough physical, endoscopical and ultrasound examination the necessity may arise to use for diagnosis angiographic and computed tomographic methods. The subject of special attention of each clinical physician should be the danger of malignant transformation of benign leiomyomatous neoplasms of the small bowel, which the authors encountered in 2 patients. PMID- 10478523 TI - [Experience of surgical treatment for stomach ulcer]. AB - The authors have analysed the results of surgical treatment of 290 patients who underwent resection of the stomach with preservation of the pylorus for ulcer disease of the stomach with its complications, as well as polyps and benign tumors of the stomach. In 57 patients combined chronic duodenal obstruction (CDO) was revealed. In 39 of them, with CDO being compensated; simultaneously the ligament of Treitz was cut and pullthrough of duodenal junction was carried out by Strong procedure, and in 18 patients with subcompensated CDO transversal antiperistaltic duodenostomy by Ya.D. Vitebsky was carried out. 12 patients with paraesophageal hiatal hernia and with gastro-esophageal reflux disease with incompetence of the cardia, simultaneously anterior crurornaphy and fundoplication by Nissen was performed. Together with conventional methods for the diagnosis of combined CDO, the disease of the liver and the bile ducts, the pancreas, hydroultrasonographic examination of the duodenum on the background of medicamentous relaxation and contrasting of the duodenum with Barium suspension is used. Long-term results from 1 to 14 years have shown efficacy of this method in 97% of patients. The absence of the duodeno-gastral reflux and relapses of gastric ulcer is due to preservation of the pyloric sphincter muscle, the effective diagnostic procedures and adequate correction of CDO. PMID- 10478524 TI - [Early reoperations after resection of stomach in gastroduodenal ulcers]. AB - The material is based on the 20 years' experience in urgent surgical treatment of 2146 patients with gastroduodenal ulcers (GDU). 120 patients of them underwent relaparotomy. The main reasons for the repeated operations were insufficiency of the sutures of the duodenal stump, peritonitis, bleeding, abscesses of the abdominal cavity. Owing to new tactics of surgical treatment for GDU the authors managed to reduce the number of relaparotomies from 9.3 to 3.7% and total mortality rate from 5 to 0.7%. PMID- 10478525 TI - [Diet regimes in acute pancreatitis]. AB - A method of determination of the duration of curative fasting in acute pancreatitis, depending on the degree of severity of patients' condition is described. The dependence between the probability of exacerbation of the process after resumption of feeding, the degree of severity of the condition at admittance, the duration of the fast vs degree of the severity of the condition in which the feeding was resumed was studied, for this purpose the method for calculation of so called coefficient of the degree of severity of acute pancreatitis was developed. 130 patients with acute pancreatitis underwent the treatment according to this method. PMID- 10478526 TI - [Heterotopic replantation of fingers after traumatic amputation]. AB - Available are the results of the treatment of 93 patients with multiple traumatic amputations of the fingers, in whom the heterotopic digital replantation was carried out. A total of 367 fingers were amputated, 187 of them have been replanted, which made up 51% of replantability. 148 fingers (79.1%) were replanted heterotopically and 39 (20.9%)--orthotopically. The viability was restored in 167 fingers, which made up 89.3%. Of 148 fingers, which have been replanted heterotopically, 88.6% have survived, and of 39 fingers, being replanted orthotopically, 92.4% survived. The analysis of functional results of the digital replantations in hand comprised the study of the hand biomechanics, blood supply and innervation. A total of 31 patients was followed up and examined in long term period from 8 months to 6 years after replantation. The biomechanics of the hand was assessed according to its capacities for seize and pinch as well as to the extent of movements in the joints of the restoration of digital innervation was assessed by the study of temperature and discrimination sensitivity. For evaluation of blood supply the US dopplerography and scintigraphy were used. The authors came to the conclusion that there was no substantial difference in the function of operated fingers both in orthotopic and heterotopic replantations. Taking into account that heterotopic replantation of the fingers resulted in more active digital function, it is safe to speak about more pronounced increase in the function of the hand in cases of heterotopic replantations of fingers in their multiple traumatic amputation. PMID- 10478527 TI - [Primary tumors of heart]. AB - The analysis of 36 years' experience in surgical treatment of 162 patients with benign and malignant primary tumors of the heart, verified by morphological examination, is presented. The possibility of their timely and intra vital revealing is noted in cases of clinical alertness which makes possible to suspect this disease and purposefully examine these patients using preferably echocardiography and computed tomography. The scope of the operations and their success depend on morphological characteristics of the tumors and the extent of neoplastic process. However, by now short- and long-term results of surgical treatment evidence, that predominant part of this category of patients is potentially curable, provided that these neoplasms of the heart were revealed at an initial stage. PMID- 10478528 TI - [Diagnosis and treatment of early bleedings after open-heart surgery]. AB - On the basis of practical experience of the clinic for Cardiovascular Surgery and up-to-date literature data the causes of early bleedings after open-heart surgery are summarized. The main clinical, roentgenological and laboratory features of the bleeding and the methods for the assessment of their origin are demonstrated. The authors consider pathophysiologic causes of hemocoagulation disturbances after surgery with artificial circulation. Characteristics of clinical and laboratory tests for coagulation and hemostasis disturbances and the algorhythms for their interpretation are described. Both nonspecific and pathogenetic methods for management of coagulopathy bleedings in early postoperative period have been described and discussed. PMID- 10478529 TI - [Combined operations for cholelithiasis]. AB - The experience of 1560 combined operations performed in N.N.Burdenko clinic from 1974 to March 1999 for cholelithiasis is analysed. The results of 7461 cholecystectomies are analysed depending on patients' age (17.8% of patients aged 65 years and older), urgency of the operations (there were 35.6% of patients of this age group who underwent surgery under strictly urgent conditions), and complicacy of the operation (combined operations were carried out in 837 patients with cholecystitis). Last years in the clinic each 10th cholecystectomy is carried out as an additional procedure in more complicated surgery, or it is combined with less complicated one. Thus, in 389 patients with more complicated diseases than cholelithiasis (approximately in a half of them tumors of various locations were observed), cholecystectomy was performed as a combined procedure (1st group), and in 448 patients (2nd group) it was the main procedure. In the 1st group 27% of patients were aged 65 years and older, the overwhelming majority of the operations were carried out by the same approach, mortality rate being 3.3%. In the 2nd group elderly patients made up 20.7%; 68.3% of patients were operated by two approaches, lethality rate being 0.2%. Thus, lethality after 837 combined operations made up 1.7%, total mortality rate after 7461 cholecystectomies made up 0.29%. PMID- 10478530 TI - Surgical treatment of diabetic osteoarthropathy, complicated by pyogenic process in the feet. AB - The data on the results of combined treatment of 540 patients with diabetic osteoarthropathies (DOAP) complicated by pyonecrotic lesions of the feet (PNLF) are presented. The authors classification of these lesions is proposed which considers location and the stage of the disease. Surgical treatment was used in 387 (71.6%) patients. The analysis of the clinical experience in surgical treatment and the results obtained in patients with DOAP and PNLF allouvs to subdivide surgical procedures in the patients in to the following categories: 1) the initial prophylaxis of development of pyonecrotic processes; 2) measures of secondary prophylaxis of the progression of pyonecrotic processes; 3) amputation of the shin and the thigh. As a result of the complex of curative and prophylactic measures, comprizing surgical treatment, the number of favourable results of treatment in patients of the lest group increased by 24%, and the number of unfavourable outcomes decreased by 25%, the mortality rate in the test group lowered 2, 4 times. PMID- 10478531 TI - [Surgical aid for population in conditions of compulsory medical insurance]. AB - A brief review of economic experiments in Russian health-protection service is carried out. It is demonstrated that their ineffectiveness is stipulated by the absence of the analysis of the causes of low surgical activity. The experiment in intensification of surgical activity, which has been carried out in Moscow hospital N 40, shous the ways for correction of present situation. An additional financing source such as CMI may substantially improve the state of surgical clinics, provided that optimal conditions for surgeons' work were created. PMID- 10478532 TI - [Choice of hemorrhoids treatment]. PMID- 10478533 TI - [Hepatobiliary surgery in 2009]. PMID- 10478534 TI - [Pilot experience of pyloroduodenal area hole closure by preparation "Takho_Comb" without preliminary suture]. PMID- 10478535 TI - [Surgical treatment of congenital biliary cysts]. PMID- 10478536 TI - [Thromboembolic complications, venous hemodynamics and lower extremities in laparoscopic operations]. PMID- 10478537 TI - [Surgery in ancient civilization of Asia]. PMID- 10478538 TI - [Maze procedure for a lone atrial fibrillation]. AB - Between 1992. 2 and 1997. 12, the maze procedure for lone fibrillation or flutter (lone af) was performed in 8 patients including 2 patients with a sustained atrial fibrillation, 5 patients with a paroxysmal atrial fibrillation and 1 patient with a sustained atrial flutter. All patients had suffered from a drug resistance lone af which induced sever symptom that is the dyspnea, palpitation and fatigue. Therefore patients requested to receive the maze procedure, and they agreed with the informed concent. After the maze operation, the normal sinus rhythm was recovered in 7 of 8 patients (87.5%). In only 1 patient, a paroxysmal atrial fibrillation remained but his symptoms improved after surgery. So this operation is a good choice of a treatment for a drug resistance lone af. PMID- 10478539 TI - [A case report of double roentgenographically occult lung cancer revealed by postsurgical pathological study]. AB - A 66-year-old man admitted to our hospital as a roentgenographically occult lung cancer (ROLC) detected by sputum cytology of class IV. A differential brushing of all branches of the bronchi was performed and squamous cell carcinoma was detected only from the right B8 segmental bronchus. Right lower lobectomy was performed and the microscopic findings of surgical specimen revealed the squamous cell carcinomas were seen at not only B8 bronchus but also B7 bronchus. The frequency of multicentricity of ROLC is reported to be high, and a differential bronchial brushing of all bronchi is a very powerful method to diagnose synchronous multiple lung cancer. However, we failed to detect a cancer lesion of B7 segmental bronchus in this case. Since, the outcomes of surgical treatments for either synchronous or metachronous multiple primary lung cancer are satisfactory, limited surgical treatments might be appropriate as an initial treatment for a ROLC. PMID- 10478540 TI - [A case of lateral femoral circumflex artery as a free graft for coronary artery bypass surgery]. AB - A 46-year-old man had a three-vessel coronary disease. We performed quadruple coronary artery bypass grafting (CABG) with the left internal thoracic artery (LITA), right gastroepiploic artery (RGEA), saphenous vein and lateral femoral circumflex artery (LFCA). Postoperative coronary angiogram showed that the LFCA bypass graft was patent and supplied sufficient blood to the anastomosed vessel. There was no stenosis at the anastomotic site. However, the LFCA graft showed a string sign. Long-term follow-up and angiographic studies is necessary to establish the use of LFCA as an arterial free graft for coronary revascularization. PMID- 10478541 TI - [New operative technique to resect of peripheral bronchogenic carcinoma with chest wall invasion using Kent's retractor]. AB - We report the new operative technique for resection of peripheral bronchogenic carcinoma with chest wall invasion using Kent's retractor and hook suspender. Standard muscle sparing thoracotomy cannot obtain the adequate operative field in the posterior chest wall under the scapula because of sparing the latissimus dorsi muscle. To obtain a sufficient operative field at the operation for en bloc chest wall resection, especially with the area of the posterior chest wall under the scapula, resection of the trapezius muscle, rhomboideus major muscle, and latissimus dorsi muscle has been added to muscle-sparing thoracotomy. Through our new technique using Kent's retractor and hook suspender that has been used in the abdominal surgery, assistant surgeon can be eliminated in favor of hanging up the scapula, which is able to be performed much easier to obtain a good operative field. To use the Kent's retractor and hook suspender, the scapula is lifted upward and cranially. With these procedures, the proximal portion of the first, second and third ribs in the posterior chest wall covering by the scapula is able to resect from vertebrocostal junction with muscle-sparing thoracotomy. We examined the apparatus for a case of the right upper lung carcinoma with invasion to the posterior chest wall. We performed right upper lobectomy with chest wall in the area of the second, third and forth ribs with suspending the scapula by Kent's retractor after muscle-sparing thoracotomy. PMID- 10478542 TI - [Efficacy of intravenous administration of atrial natriuretic peptide after cardiac surgery in neonates and infants]. AB - Intravenous infusion of alpha-human atrial natriuretic polypeptide (hANP) and furosemide was performed in 12 patients (4: TGA, 3: univentricular heart, 3: HLHS, 2: VSD) after cardiopulmonary bypass. Their mean age at the operation was 68 days (6 patients in neonate), the mean body weight was 3.3 kg. All patients were treated with nitroglycerin in dose of 2 micrograms/kg/min and chlorpromazine in dose of 2.8 micrograms/kg/min and catecholamine in mean dose of 6.5 micrograms/kg/min. The criteria of indication for hANP was poor effect of furosemide alone. The hANP was given for 6-144 hours in dose of 0.1-0.2 microgram/kg/min. With the hANP and furosemide infusion, urine volume increased from 85.0 +/- 14.2 ml/kg/day to 107.9 +/- 25.3 ml/kg/day (p < 0.05), whereas the systemic arterial pressure, the central venous pressure and the renal function were unchanged. We conclude that the combination of the hANP and the furosemide is very effective in neonates and infants. PMID- 10478543 TI - [Is left ventricular regional wall motion after CABG influenced by cardioplegic protection?]. AB - This clinical study was conducted to determine whether different techniques of cardioplegic protection reflected left ventricular regional wall motion after CABG. A total of 43 patients with more than 90% stenosis of the LAD (seg 6 and/or 7) before CABG, who had patent grafts were allocated to two groups: namely, the crystalloid group, comprised of 23 patients given cold crystalloid cardioplegia and topical ice slush, and the blood group, comprised of 20 patients given tepid blood cardioplegia delivered intermittently antegrade. Each group was divided into two subgroups according to whether the left ventricular regional wall motion showed no change or deterioration after CABG. We also examined the relationship between the grading of the collateral artery before CABG and the postoperative ventricular regional wall motion. The number of patients who showed deterioration after CABG was higher in the crystalloid group than in the blood group (p = 0.008). Moreover, patients in the crystalloid group whose collateral artery had been graded as 0 before CABG tended to show deterioration of left ventricular local wall motion after CABG (p = 0.07). Whereas those patients in the blood group did not. In conclusion, the incidence of deterioration after CABG was higher in the crystalloid group than in the blood group. PMID- 10478544 TI - [Efficacy of terminal warm blood cardioplegia in combination with Bretschneider HTK solution for myocardial protection]. AB - Many reports indicated that terminal warm blood cardioplegia (TWBC) was useful for reduction of the reperfusion injury in cardiac operations. We introduced Bretschneider-HTK solution (B-HTK) from April 1992, and combined use of the TWBC and the B-HTK was performed from May 1996. The purpose of this study was to evaluate clinical efficacy of the TWBC in combination with B-HTK for myocardial protection. In patient with valve operations from May 1995 to May 1997, clinical results were compared between the B-HTK with TWBC group (34 patients) and the B HTK alone group (37 patients). Between two groups, there was no significant difference in patient characteristics, preoperative NYHA class, cardiopulmonary bypass time, aortic cross clamp time, minimum rectal temperature, rewarming and circulatory support time, incidence of IABP and blood concentration of max CPK, max GOT. Furthermore, the cardiac index in 1 postoperative day showed some trend to be higher in the combined TWBC group than that in the B-HTK alone group. But there was no significant difference between two groups in the LV shortening fraction at 14 postoperative day. Incidence of DC defibrillation was significantly lower in the combined TWBC group than that in the B-HTK alone group. These results suggested that combination of the TWBC might be useful for reduction of the reperfusion injury following myocardial protection with B-HTK solution. PMID- 10478545 TI - [Clinical comparison of diffuse malignant mesothelioma of the pleura and pseudomesotheliomatous carcinoma of the lung for each case]. AB - Because we experienced each 1 operative case of diffuse malignant mesothelioma of the pleura and pseudomesotheliomatous carcinoma of the lung discovered with pleural effusion, clinical comparison investigated both. The first case was suspected diffuse malignant mesothelioma of the pleura before operation, and we performed pleuropneumonectomy. But the pathologic diagnosis was adenocarcinoma of the lung, what is so called pseudomesotheliomatous carcinoma. The second case had inhaled asbestos and his pleural effusion revealed high concentrations of hyaluronic acid. Thoracoscopic biopsy showed malignant mesothelioma, and we performed pleuropneumonectomy. The pathologic final diagnosis was diffuse malignant mesothelioma of the pleura. In clinical differential diagnosis of diffuse malignant mesothelioma of the pleura and pseudomesotheliomatous carcinoma of the lung, history of inhalation of asbestos and concentrations of hyaluronic acid in pleural effusion are helpful. And thoracoscopic biopsy is necessary in established diagnosis. PMID- 10478546 TI - [Progressive mitral regurgitation which is necessitated the mitral valve replacement after partial left ventriculectomy (Batista procedure): a case report]. AB - The patient is 61-year-old woman who underwent partial left ventriculectomy, (Batista procedure) due to dilated cardiomyopathy and multiple thromboembolism. Although postoperative course was uneventful, she has had clinical symptoms of the left heart failure due to the increased mitral valve regurgitation at the early postoperative period, gradually. Even though mitral valve regurgitation was severe, it was not apt to re-dilatate the left ventricular capacity evaluated by echocardiography. She underwent the mitral valve replacement on the 92nd postoperative day, and was once possible for weaning from cardiopulmonary bypass under the support of IABP. However, she died on the 19th postoperative day caused by sepsis. It is important to evaluate the accurate mitral valve regurgitation preoperatively for Batista procedure. Although there was the mild mitral valve regurgitation, it is essential to repair or replace the mitral valve for Batista procedure. PMID- 10478547 TI - [A case of large congenital right ventricular outflow aneurysm]. AB - A rare case of large congenital right ventricular outflow aneurysm is reported. A 5-year-old girl had been diagnosed as having atrial septal defect and pulmonary valvular stenosis. A large cavity in front of right ventricular outflow was noticed by her follow-up echocardiography. The non-contracting cavity was confirmed as an aneurysm originating from right ventricular outflow tract by angiocardiogram. Surgical resection was performed successfully with concomitant atrial septal defect closure and pulmonary valvular commissurotomy. Histological examination revealed that the wall of the ventricular outflow aneurysm consisted of dense fibrous tissue, contained no myocardium, and the cavity was lined with endocardium. It is reported that rupture of congenital ventricular aneurysm may occur even if there is no symptom. Surgical resection will be recommended especially when increasing in size is diagnosed. PMID- 10478548 TI - [Double chambered right ventricle in a 62-year-old female: a surgical case report]. AB - A 62-year-old female was admitted with a chief complaint of transient syncope on exertion. Angiography in right ventricle revealed a defect caused by an anomalous muscle bundle and a pressure gradient of 151 mmHg was observed between the two chambers by cardiac catheterization. Resection of the anomalouse muscle bundle was undertaken using a lower median sternotomy starting at the 2nd intercostal space level and through the outflow tract right ventriculotomy. Patch plasty was also undertaken in the outflow tract. Post-operative course was uneventful and pressure gradient had disappeared at the post-operative catheterization. A rare case of DCRV in a 62-year-old patient with a pressure gradient of 151 mmHg in the right ventricle was reported. PMID- 10478549 TI - [A case of left atrial thrombus complicated with coarctation of the aorta]. AB - The case was a sixty one year old female who was diagnosed as having acute arterial embolism of right upper arm and underwent embectomy. The detailed examination which followed revealed left atrial myxoma and coarctation of the aorta. Due to the risk of embolism, an emergency operation was performed to install a right axillofemoral bypass with an 8 mm artificial graft. Left atrial myxoma was then excised in an operation using an artificial heart-lung apparatus. Pathological examinations of excised tumor offered no trace of tumor but only of left atrial thrombus. Postoperative results were satisfactory. The pressure difference between upper and lower arms was improved to 0.8 in pressure index from preoperative 0.5 and it became possible to control the blood pressure of upper arm within the level of 130 to 150 mmHg. Axillofemoral bypass is an effective method for a case like the above where a site of coarctation of the aorta cannot be directly repaired. PMID- 10478550 TI - [Reoperation of tetralogy of Fallot for the transannular patch neo-intimal stenosis: a case report]. AB - A 3-year-old male patient underwent right ventricular outflow tract reconstruction with a glutaraldehyde-preserved equine pericardium for tetralogy of Fallot. Because of progressive severe pulmonary restenosis with over systemic right ventricular pressure, tricuspid regurgitation, and abnormal high echoic shadow in the distal main pulmonary artery on echocardiogram, he required reoperation a year after the first correction. In the reoperative findings, the pseudointima was thickened heavily and detached from glutaraldehyde-preserved equine pericardial patch. The patch was removed and the right ventricular outflow was reconstructed widely to the pulmonary bifurcation with porcine pericardial patch again. Patho-histological findings showed foreign body giant cells and macrophages in the pseudointima. Four years after the reoperation, echocardiogram shows 41 mmHg for the right ventricular pressure and 22 mmHg for the pressure gradient of right ventricular outflow tract, and the patient is doing well now. PMID- 10478551 TI - [A case of successful palliative operation of asplenia syndrome with total anomalous pulmonary venous return in young infant]. AB - We reported a successful palliative operation for asplenia syndrome with total anomalous pulmonary venous return (TAPVR Ia) in an infant. The boy was suffering from cyanosis and tachypnea. He was diagnosed as asplenia syndrome with TAPVR and hiatus hernia. After he was admitted to our hospital, pulmonary congestion gradually progressed in a month. At 58 days of age, a palliative operation (repair of TAPVR and pulmonary artery banding with band of 20 mm in length) was performed. The postoperative course was uneventful. At 114 days of age, he underwent curative operation for hiatus hernia without cardiac failure. Postoperative cardiac catheterization at 179 days of age showed appropriate pulmonary artery pressure. We emphasize that pulmonary artery banding which is tighter than usual well controls pulmonary blood flow, although the length of the band in each case should be considered individually. PMID- 10478552 TI - [Two case reports of bilateral synchronous pneumothorax treated in a single stage bilateral thoracoscopic surgery]. AB - We report here two cases of bilateral synchronous pneumothorax treated by video assisted thoracoscopic surgery. The first case was a 15-year-old male and the second case was a 22-year-old male. Bilateral thoracoscopic surgery by using of endo-stapler was performed at one stage and Nd-YAG Laser ablation to both the emphysematous surroundings and multiple small blebs was added. This procedure is minimally invasive as well as effective for the treatment of bilateral synchronous pneumothorax. PMID- 10478553 TI - [A case of mediastinal myelolipoma]. AB - Myelolipoma is a benign tumor composed of mature adipose tissue and hematopoietic tissue. It was mainly found in the adrenal grand, but there have been reports of extra-adrenal locations. Only 7 cases of mediastinal myelolipoma have been reported ever. We have experienced a case of mediastinal myelolipoma surgically resected from a 55-year-old-man. He has visited our department because of chest pain. Chest X-ray showed typical pneumothorax, but abnormal mediastinal shadow was remarked. Chest CT showed a well circumscribed mass with fat and soft-tissue attenuation in posterior mediastinum. Lung bullectomy and mediastinal tumor resection under VATS was performed. A blue-red tumor 4 cm in diameter was resected en bloc. Pathological examination showed its composition of mature adipose tissue and hematopoietic tissue. Diagnosis was extra-adrenal myelolipoma. No recurrent nor abnormality was found in 8 months since surgery. PMID- 10478555 TI - [A case of thymic carcinoid with multiple endocrine neoplasm (MEN)-type I]. AB - We presented a case of thymic carcinoid with MEN type I. A 43-year-old woman who followed at MEN type I for 4 years was pointed out an abnormal shadow by chest X ray. Chest CT levealed the presence of two anterior mediastinal tumors. Extended total thymectomy was performed through the median sternotomy. There were 3 tumors in thymus. Histological examination revealed three tumors in the thymus and all of the tumor were diagnosed carcinoid. Our experience suggests that CT or MRI of the chest should be considered as part of clinical screening in patients with MEN type I. PMID- 10478554 TI - [An effective case of a new biological adhesive agent, gelatin-resorcinol formaldehyde-glutaralhyde glue (GRFG-glue) in treating refractory pulmonary fistula following lobectomy for pulmonary aspergilloma]. AB - We report a successful case of thoracoscopic therapy using a new biological adhesive agent, Gelatin-Resorcinol Formaldehyde glue (GRFG glue) for refractory pulmonary fistula. A 69-year-old male underwent right upper lobectomy for lung aspergilloma. Air leakage began 11 days after lobectomy. Closing alveolar fistula was performed 28 days after first operation. Relapsing air leakage began 2 days after second operation. The insertion of fibrin glue through thoracoscope at two times was not effective for refractory pulmonary fistula. But the insertion of GRFG glue was effective to close the fistula completely. PMID- 10478556 TI - [Cases of sternal metastasis except from breast cancer]. AB - Surgical resection is the option for sternal metastasis from breast cancer. But surgical treatment is the subject of controversy for sternal metastasis except from breast cancer. Four cases of secondary malignant tumors of the sternum except from breast cancer were reported. Primary lesions included thyroid cancer, gastric cancer, malignant fibrous histiocytoma of the chest wall and multiple myeloma. In 3 cases, operation were done and they survived 11 months, 13 months and 3 years respectively. In case of secondary malignant tumor of the sternum except from breast cancer, surgical treatment should be considered. PMID- 10478557 TI - [A case of superior mediastinal neurinoma resected from the supra-clavicular area]. AB - We report a case of superior mediastinal neurinoma that was resected from the supra-clavicular area. An abnormal shadow on the chest X-ray film was pointed out in a 53-year-old male. A left supra-clavicular mass was subsequently also discovered. We underwent extirpation of the tumor through supra-clavicular approach and confirmed to be a neurinoma histologically. We consider supra clavicular approach which allows a good view might be useful. PMID- 10478558 TI - Low-affinity NMDA receptor channel blockers inhibit acquisition of intravenous morphine self-administration in naive mice. AB - Experimental evidence suggests that NMDA receptor antagonists modulate behavioral effects of morphine in models assessing abuse potential of drugs. The present study sought to evaluate the ability of NMDA receptor channel blockers to affect the acquisition of morphine i.v. self-administration in drug- and experimentally naive mice. DBA/2 mice were allowed to self-administer morphine (0.125-4.0 mg/ml) or saline during the 30-min test. Each nose-poke of the active mouse resulted in a 1.6-microl infusion to both the active mouse and the passive (yoked control) mouse. In vehicle-treated mice, differences between operant activity of active and passive mice were most obvious when active mice were allowed to self-inject morphine at the concentration of 0.5 mg/ml (the optimum concentration). Pretreatment with MRZ 2/579 (1-amino-1,3,3,5,5-pentamethyl-cyclohexan hydrochloride; 1, 3.2 and 10 mg/kg) shifted the optimum concentration to 0.75 mg/ml. Memantine (1-amino-3,5-dimethyladamantane hydrochloride; 0.3, 1, 3.2 and 10 mg/kg) suppressed both the morphine intake and the difference in nose-poke activity of active vs. passive mice across all tested concentrations of morphine. Dizocilpine ((+)-5-methyl-10,11-dihydro-5H-dibenzocyclohepten-5,10-imine maleate; 0.1 mg/kg) was ineffective. Taken together with earlier reports, the present results suggest that low-affinity NMDA receptor channel blockers--in contrast to dizocilpine--attenuate the rewarding potential of morphine. PMID- 10478559 TI - Site-specific NMDA receptor antagonists produce differential effects on cocaine self-administration in rats. AB - The effects of site-specific NMDA receptor antagonists on intravenous cocaine self-administration were examined in rats trained to self-administer cocaine (0.25 mg/infusion) on a fixed ratio (FR) 5 schedule with a 20-s time-out (TO) after each reinforcer. The non-competitive NMDA receptor antagonists, dizocilpine (MK-801, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate) (0.05-0.2 mg/kg i.p.) and memantine (1,3-dimethyl-5-amino adamantane hydrochloride) (2.5-20 mg/kg i.p.), dose-dependently decreased cocaine self-administration, while the competitive NMDA receptor antagonist, CGP 39551 (DL-(E)-2-amino-4-methyl-5-phosphono-3-pentanoic acid carboxyethylester) (2.5-15 mg/kg i.p.), and the NMDA/glycine receptor antagonist, L-701,324 (7-chloro-4 hydroxy-3(3-phenoxy)-phenyl-2(H)quinolone) (1.25-10 mg/kg p.o.), were without effect. Under a progressive ratio (PR) schedule, dizocilpine (0.15 mg/kg i.p.) increased the number of cocaine infusions in a manner similar to increasing the unit dose of cocaine, suggestive of potentiation of cocaine reward. Conversely, memantine (10 mg/kg i.p.) produced rate-decreasing effects on the PR schedule. These results demonstrate that NMDA receptor antagonists acting at different modulatory sites of the NMDA receptor do not share dizocilpine's cocaine reward enhancing effects although they are all known to be effective blockers of NMDA receptor activity. PMID- 10478560 TI - Intravesical resiniferatoxin desensitizes rat bladder sensory fibres without causing intense noxious excitation. A c-fos study. AB - In this study the desensitizing power of increasing concentrations of resiniferatoxin applied topically to the bladder mucosa, and the irritating properties of the most effective desensitizing dose, were determined with the aid of the spinal expression of the proto-oncogene c-fos. Desensitization was assessed by the decrease in the number of Fos-immunoreactive spinal neurons induced by the intravesical instillation of 1% acetic acid, when the latter was preceded by resiniferatoxin in concentrations between 1 and 1000 nM. Irritation, as shown by the noxious excitation of vesical sensory innervation, was measured by the c-fos response evoked by a single application of resiniferatoxin. As to the desensitizing power, resiniferatoxin produced a dose-dependent effect with a maximum at 100 nM, which decreased Fos-immunoreactive cell numbers to less than 10% of controls. No further decrease of c-fos activation occurred at 1000 nM. As to the irritating power, the saturation dose of resiniferatoxin (100 nM) produced a very weak c-fos activation in lumbosacral spinal cord segments. These data show that in an effective desensitizing concentration, resiniferatoxin is virtually devoid of nociceptive effects, in agreement with current clinical observations. PMID- 10478561 TI - Effects of conditioned fear stress on serotonin neurotransmission and freezing behavior in rats. AB - In an attempt to clarify the role of the brain serotonergic system in the psychopathology of anxiety, we examined the effect of a psychological stress, conditioned fear stress, on extracellular serotonin (5-hydroxytryptamine, 5-HT) concentrations in the rat medial prefrontal cortex using the method of in vivo microdialysis, while simultaneously observing conditioned fear stress-induced freezing behavior, an index of anxiety. Conditioned fear stress increased extracellular 5-HT levels in the medial prefrontal cortex, and this 5-HT level increase was followed by a resolution of the freezing behavior. A dose of 10 mg/kg of a selective 5-HT reuptake inhibitor, citalopram, administered 60 min before exposure to conditioned fear stress increased extracellular 5-HT concentrations immediately and potently, reducing freezing behavior. These findings strongly suggest that facilitation of brain 5-HT neurotransmission decreases anxiety, which is in agreement with the clinical reports that selective 5-HT reuptake inhibitors are effective in the treatment of anxiety disorders. PMID- 10478562 TI - Electrophysiological effects of E-5842, a sigma1 receptor ligand and potential atypical antipsychotic, on A9 and A10 dopamine neurons. AB - Extracellular single unit recording techniques were used to study the effects of the novel potential atypical antipsychotic E-5842, (4-(4-fluorophenyl)-1,2,3,6 tetrahydro-1-[4-(1,2,4-triazol-1-il)bu tyl]pyridine citrate), a preferential sigma1 receptor ligand, on the activity of dopamine cells in substantia nigra pars compacta (A9) and ventral tegmental area (A10) in anesthetized rats. Acute i.v. administration of E-5842 (up to 3.2 mg kg(-1)) did not change the spontaneous activity of the dopamine neurons, which still responded to the inhibitory effect of a subsequent administration of high dose of apomorphine. Acute administration of E-5842 (20 mg kg(-1), i.p.) did not change the number of spontaneously active A9 or A10 dopamine cells. Chronic administration of E-5842 (20 mg kg(-1) day(-1) x 21 days, s.c.) decreased the number of spontaneously active A10 but not A9, dopamine neurons. This effect was reversed by the administration of apomorphine, thus, indicating a possible depolarization inactivation phenomenon. Our results suggest an influence of E-5842 on dopaminergic neurotransmission, although the exact mechanism remains unknown. The effect of E-5842 on A10 is similar, in some ways, to the effects observed with several atypical antipsychotics and suggest the atypicality of the compound and that E-5842 may exert its antipsychotic effects without causing significant extrapyramidal side effects. PMID- 10478563 TI - Protective effect of imidaprilat, a new angiotensin-converting enzyme inhibitor against 1-methyl-4-phenylpyridinium ion-induced *OH generation in rat striatum. AB - We examined the antioxidant effects of angiotensin-converting enzyme inhibitor on 1-methyl-4-phenylpyridinium ion (MPP+)-induced hydroxyl radical (*OH) formation in extracellular fluid of rat striatum. Rats were anesthetized and sodium salicylate in Ringer's solution (0.5 nmol microl(-1) min(-1) was infused through a microdialysis probe to detect the generation of *OH, as reflected by the non enzymatic formation of 2,3-dihydroxybenzoic acid (DHBA) in the striatum. MPP+ clearly produced an increase in *OH formation in a concentration-dependent manner. When imidaprilat was infused in MPP+ -pre-treated animals, the formation of dopamine and 2,3-DHBA significantly decreased, as compared with that in the MPP+ -only-treated group. We compared the ability of two non-SH-containing angiotensin-converting enzyme inhibitors (imidaprilat and enalaprilat) with an SH containing angiotensin-converting enzyme inhibitor (captopril) to scavenge *OH. All three angiotensin-converting enzyme inhibitors were able to scavenge *OH generated by the action of MPP+. However, the changes produced by captopril and enalaprilat were not significant. When dopamine was administered to the MPP+ -pre treatment group, a marked elevation was observed, showing a positive linear correlation between dopamine and *OH formation (2,3-DHBA) in the dialysate. Moreover, when iron (II) was administered to the MPP+ -pre-treatment group, the same results were obtained: a positive linear correlation (R2 = 0.989) between the release of dopamine and 2,3-DHBA (R2 = 0.989) in the dialysate. When corresponding experiments were performed with imidaprilat-pre-treated animals, the level of 2,3-DHBA decreased. These results suggested that angiotensin converting enzyme inhibitors may protect against MPP+ -induced *OH formation in the rat striatum. PMID- 10478564 TI - Influence of isoprostanes on vasoconstrictor effects of noradrenaline and angiotensin II. AB - The isoprostanes, 8-iso-prostaglandin F2alpha and 8-iso-prostaglandin E2, which are released in vivo by free radical-catalyzed peroxidation of arachidonic acid, are potent vasoconstrictors. Increased formation of 8-iso-prostaglandin F2alpha has been detected in human cardiovascular diseases, in which enhanced plasma levels of noradrenaline and angiotensin II have harmful vasoconstrictor effects. Therefore, we investigated the influence of perfusions with the thromboxane A2 mimetic, U 46619, and with the isoprostanes, 8-iso-prostaglandin F2alpha, 8-iso prostaglandin E2, 8-iso-prostaglandin E1 and 8-iso-prostaglandin F3alpha, on the vasoconstrictor effects of noradrenaline and angiotensin II in the isolated perfused rabbit ear. Our results demonstrate that perfusions with U 46619, 8-iso prostaglandin E2 and 8-iso-prostaglandin F2alpha, at a subthreshold concentration (30 nM), amplified the vasoconstrictions induced by noradrenaline or angiotensin II significantly. In addition, the results show that U 46619, 8-iso-prostaglandin F2alpha, 8-iso-prostaglandin E2 and 8-iso-prostaglandin E1, which were applied as a bolus, induced much more pronounced vasoconstrictions than prostaglandin F2alpha, prostaglandin E2 and prostaglandin F3alpha. Prostaglandin E1 and 8-iso prostaglandin F3alpha, showed no effects. In conclusion, it can be assumed that the powerful vasoconstrictions induced by 8-iso-prostaglandin E2 and 8-iso prostaglandin F2alpha and their potentiating effects on vasoconstrictions induced by noradrenaline or angiotensin II might be of pathophysiological relevance in cardiovascular diseases. PMID- 10478565 TI - Progesterone modulates estradiol actions: acute effects at physiological concentrations. AB - The progestin element of hormone replacement therapy may reduce the cardioprotective actions of the estrogen component. Only high concentrations (microM) of progesterone directly relaxed U46619 (9,11-dideoxy-9alpha, 11alpha methanoepoxy prostaglandin F2alpha)-pre-contracted porcine coronary artery rings. A low concentration of progesterone (1 nM), with no effects of its own, shifted the relaxation curves of bradykinin and calcium ionophore A23187 to the right while not affecting those of sodium nitroprusside and levcromakalim. The negative influence that 1 nM progesterone exerted on bradykinin- and A23187-mediated relaxation was diminished when 1 nM 17beta-estradiol was concomitantly added to the bathing medium. Conversely, the potentiating actions of 1 nM 17beta-estradiol on relaxations elicited by sodium nitroprusside and levcromakalim were reduced following simultaneous treatment with the same concentrations of progesterone. These findings represent the first evidence for an acute in vitro vascular effect of progesterone at a physiologically relevant concentration and concur with previous in vivo reports demonstrating that progesterone may diminish the beneficial effects of estrogens. PMID- 10478566 TI - The effect of cis-Diamminedichloroplatinum II on Na+ and K+ transport in the rabbit cortical collecting duct. AB - cis-Diamminedichloroplatinum II (CDDP) is an antineoplastic drug against solid malignant tumors. However, its clinical use is limited by nephrotoxicity. CDDP also causes hypokalemia and in vivo microperfusion method have demonstrated that luminal CDDP increases K+ secretion by hyperpolarization of the transepithelial voltage difference through stimulating Na+ transport in the distal segments. However, there is no direct evidence for this mechanism. We therefore examined the effect of luminal CDDP on Na+ and K+ transport in the rabbit cortical collecting duct (CCD) using in vitro isolated tubular microperfusion. Luminal CDDP hyperpolarized the transepithelial voltage difference (V(T)) in a dose dependent manner at concentrations from 10(-5) M to 10(-3) M and at 10(-3) M CDDP, V(T) was hyperpolarized from -11.6+/-2.3 mV to -16.6+/-3.3 mV (P<0.001). A concentration of 10(-5) M ouabain, 10(-4) M amiloride and 2 mM BaCl2 all completely abolished CDDP-induced hyperpolarization. To confirm the mechanism, Na+ and K+ flux were measured in the presence of 10(-3) M CDDP. CDDP decreased net K+ secretion from -22.2+/-5.7 to -15.2+/-2.9 pmol mm(-1) min(-1) (P<0.01) without any effect on the lumen-to-bath isotope flux of Na+ (52.6+/-10.6 to 52.1+/-10.7 pmol mm(-1) min(-1)). These data suggest that luminal CDDP hyperpolarizes V(T) primarily by inhibiting K+ conductance but did not influence Na+ transport of the luminal membrane. We conclude that the CCD does not play a role in CDDP-induced hypokalemia when CDDP is applied from the luminal side. PMID- 10478568 TI - Relaxant activity in rat aorta and trachea, conversion to a muscarinic receptor antagonist and structure-activity relationships of new K(ATP) activating 6-varied benzopyrans. AB - To characterize ATP-sensitive channels (K(ATP) channels) benzopyrans with different substituents at position 6 were synthesized as new K(ATP)-activators. Their relaxant potencies were determined in rat aorta and trachea. In aorta, pEC50-values (-log, M) ranged from 7.37 to 5.43; in trachea, pEC50-values were 0.3 to 0.8 log units lower. Functional data were compared with binding data obtained in calf tracheal cells using the cyanoguanidine [3H]P1075 (N-cyano-N' 1,1-dimethyl[2,3(n)-3H]propyl)-N11-(3-pyridinyl)guanidine) as radioligand. A high correlation (r = 0.96) between pEC50- and pKD-values indicated that tracheal relaxation produced by benzopyrans is mediated via K(ATP) channels without signal amplification. The permanently charged trimethylammonium derivative designed as a probe for the membrane site of action completely lost its affinity for K(ATP) channels, but converted to an antagonist for muscarinic acetylcholine receptors (pK(B) = 6.12+/-0.10), as confirmed in radioligand binding studies (pK(D) = 5.77+/-0.04). Structure-activity analyses revealed that the 6-substituent influences biological activity by a direct receptor interaction of its own and not indirectly by withdrawing electrons from the benzopyran nucleus. The variance of the biological activity is primarily determined by electrostatic properties, but desolvation energies additionally contribute. PMID- 10478567 TI - Buspirone functionally discriminates tissues endowed with alpha1-adrenoceptor subtypes A, B, D and L. AB - The affinity for functional alpha1-adrenoceptor subtypes of buspirone in comparison with its close structural analogs and selective alpha1D-adrenoceptor antagonists, BMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8 azaspiro[4.5]dec ane-7,9-dione) and MDL 73005EF (8-[2-(1,4-benzodioxan-2 ylmethylamino)ethyl]-8-azaspiro+ ++[4.5]decane-7,9-dione), was determined, namely at subtype A in rat vas deferens and perfused kidney, at subtype B in guinea-pig and mouse spleen, at subtype L in rabbit spleen, and at subtype D in rat aorta and pulmonary artery against noradrenaline-evoked contractions. BMY 7378 and MDL 73005EF were confirmed as 30- and 20-fold selective antagonists, respectively, for alpha1D- over both alpha1A- and alpha1B-adrenoceptors. Buspirone was a weak antagonist without intrinsic activity at alpha1A-adrenoceptors in rat vas deferens (pA2 = 6.12), at alpha1B-adrenoceptors in guinea-pig and mouse spleen (pA2 = 5.54 and 5.59) and at alpha1L-adrenoceptors in rabbit spleen (pA2 = 4.99), but caused partial vasoconstriction in rat kidney that was attenuable by the subtype D-selective adrenoceptor antagonist BMY 7378, but hardly by the subtype A selective adrenoceptor antagonist B8805-033 ((+/-)-1,3,5-trimethyl-6-[[3-[4-((2,3 dihydro-2-hydroxymethyl)-1,4-be nzodioxin-5-yl)-1-piperazinyl]propyl]amino] 2,4(1H,3H)-pyrimidinedion e), confirming the additional presence of alpha1D adrenoceptors mediating rat renal vasoconstriction. Buspirone behaved as a partial agonist at alpha1D-adrenoceptors in rat aorta (pD2 = 6.77, intrinsic activity (i.a.)= 0.40) and pulmonary artery (pD2 = 7.16, i.a. = 0.59). With buspirone as agonist in these tissues, the pA2 values of subtype-discriminating antagonists were consistent with their alpha1D-adrenoceptor affinity determined in rat aorta against noradrenaline and with published binding data on cloned alpha1d-adrenoceptors. The results provide pharmacological evidence that (1) in functional preparations for the A subtype, like rat vas deferens and perfused kidney, for the B subtype, like guinea-pig and mouse spleen, and for the L subtype, like rabbit spleen, buspirone is a weak antagonist without intrinsic activity, but (2) behaves as a partial agonist in rat aorta and pulmonary artery as models for the D subtype and (3) detects an additional vasoconstrictor alpha1D adrenoceptor in rat kidney. Buspirone, like its close analogs BMY 7378 and MDL 73005EF, thus might also be a useful tool for functionally discriminating alpha1D from alpha1A-, alpha1B- and alpha1L-adrenoceptors in various tissues. PMID- 10478569 TI - Inhibitory prostanoid EP receptors in human non-pregnant myometrium. AB - Prostanoid EP receptor agonists relaxed cloprostenol-stimulated contraction of human non-pregnant myometrium in vitro with pEC50 values of (n = 4): prostaglandin E2, 7.8+/-0.2 > 1-OH prostaglandin E1, 7.2+/-0.3 > misoprostol, 6.6+/-0.1 > 16,16-dimethyl prostaglandin E2, 6.3+/-0.7 > butaprost, 5.7+/-0.3 > 11-deoxy prostaglandin E1, 5.5+/-0.2 = AH13205 ((+)-trans-2-[4 (1hydroxyhexyl)phenyl]-5-oxocyclopentaneheptano ic acid), 5.5+/-0.2. The EP4 receptor antagonist AH23848B ([1alpha(z), 2beta5alpha]-(+/-)-7-[5-[[(1,1' biphenyl)-4-yl]methoxy]-2-(4-morph olinyl)-3-oxo-cyclopentyl]-4-heptenoic acid) (29 microM) had no effect on concentration-effect curves to the EP receptor agonists. The mixed prostanoid receptor antagonist AH6809 (6-isopropoxy-9 oxaxanthene-2-carboxylic acid) competitively antagonised prostaglandin E2 with a pA2 of 5.6+/-0.2. AH6809 (42 microM) antagonised misoprostol, 11-deoxy prostaglandin E1, and the prostanoid DP receptor agonist BW245C (5-(6 carboxyhexyl)-1-(3-cyclohexyl-3-hydroxypropyl)hydantoin) with apparent pA2 values of 5.6+/-0.3, 5.1+/-0.9 and 5.9+/-0.4 (n = 4), respectively, but was ineffective against the IP receptor agonist cicaprost (n = 4). The prostanoid DP receptor antagonist BW A868C (3-benzyl-5-(6-carboxyhexyl)-1-(2-cyclohexyl-2 hydroxyethylamino)h ydantoin) (50 nM) had no effect on responses to prostaglandin E2 or misoprostol. The presence of an AH6809-sensitive, AH23848B- and BW A868C insensitive mechanism is consistent with the hypothesis that inhibitory EP receptor agonists cause relaxation of human non-pregnant myometrium by an EP2 receptor-mediated process. PMID- 10478570 TI - Impairment of epithelium-dependent relaxation in coaxial bioassay by reactive oxygen species. AB - The purpose of the present study was to investigate the effects of reactive oxygen species on the activity of epithelium-derived relaxant factor (EpDRF) released by guinea-pig tracheal epithelium. Reactive oxygen species were generated by the electrolysis of the physiological buffer in which the tissues were bathed. Epithelium-dependent relaxation induced by acetylcholine in precontracted rat anococcygeus muscle that was placed in epithelium-intact guinea pig trachea (coaxial bioassay system) was significantly attenuated when the tissues were exposed to electrolysis. Impairment of the acetylcholine response was prevented by incubation with free radical scavengers prior to electrolysis. In isolated rings of guinea-pig trachea, the contractile responses elicited by acetylcholine, histamine and 5-hydroxytryptamine were not altered after electrolysis of the bathing solution. The results of the present study suggested that exposure to reactive oxygen species impaired EpDRF release from guinea-pig trachea epithelium but did not alter the contractility of tracheal smooth muscle. PMID- 10478571 TI - Beta-adrenoceptor reserve in human lung: a comparison between airway smooth muscle and mast cells. AB - The effects of several beta-adrenoceptor agonists on the relaxation of precontracted human bronchial rings and the inhibition of IgE-mediated histamine release from human lung mast cells (HLMC) were studied. For the relaxation of bronchial rings, isoprenaline, fenoterol and terbutaline were full agonists whereas salbutamol was a full agonist in some (two out of six) experiments and a partial agonist in the remainder. For the inhibition of histamine release, relative to isoprenaline, neither fenoterol, terbutaline nor salbutamol was a full agonist. Studies with the irreversible beta-adrenoceptor antagonist, bromoacetylalprenolol menthane, indicated that there was a larger beta adrenoceptor reserve for the relaxation of precontracted bronchial rings than for the inhibition of histamine release from HLMC. Further studies indicated that the isoprenaline inhibition of histamine release was considerably more susceptible to desensitizing treatments than the isoprenaline relaxation of bronchial rings. Collectively, these data suggest that a larger beta-adrenoceptor reserve exists for the relaxation of smooth muscle than the inhibition of histamine release from HLMC and that differences in receptor reserve may contribute to the relative susceptibilities of the two systems to desensitization. PMID- 10478572 TI - Effects of nitric oxide donors GEA 3162 and SIN-1 on ethanol-induced gastric ulceration in rats. AB - Low doses of the intragastrically (i.g.) administered nitric oxide (NO) donors, 1,2,3,4-oxatriazolium,5-amino-3-(3,4-dichlorophenyl)-chloride (GEA 3162; 0.3 mg/kg) and 3-morpholino-sydnonimine (SIN-1; 1 mg/kg), inhibited gastric ulceration induced by ethanol (94%) in anesthetized rats. In contrast, higher doses of these NO donors administered i.g. exacerbated the damage. When administered intravenously, the NO donors had no effect on ethanol-induced gastric lesions although a clear blood pressure-lowering effect was seen. Neither the inhibition nor the exacerbation of ulceration was correlated with changes in blood pressure or prostaglandin E2 release from the mucosal tissue. The relatively small difference between the gastroprotective and damaging doses suggests that orally administered NO donors, especially in the case of GEA 3162, may have a narrow gastric safety margin. PMID- 10478573 TI - Strong antiproliferative effects of baicalein in cultured rat hepatic stellate cells. AB - Recently, antifibrogenetic effects of Sho-saiko-to, a traditional herbal medicine in Japan, have been shown in experimental hepatic fibrosis, and flavonoids in Sho saiko-to are suspected as active ingredients. Thus, we evaluated the effects of baicalein, a major flavonoid in Sho-saiko-to, on proliferation and protein synthesis in cultured rat hepatic stellate cells. Baicalein decreased [3H]thymidine incorporation in cells stimulated with platelet-derived growth factor-B subunit homodimer (PDGF-BB) in a concentration-dependent manner (approximate ED50<10 microM, P<0.0001), and the decrease observed with 10 microM baicalein was greater than those observed with 5 microM retinol or 500 microM 3 isobutyl-1-methylxanthine (IBMX). Baicalein consistently decreased [3H]thymidine incorporation and cell number in cells stimulated with fetal calf serum (ED50<10 microM, P<0.0001), and moderately suppressed [3H]leucine and [3H]proline incorporation (P<0.0001). These results demonstrate the strong antiproliferative effect of baicalein in hepatic stellate cells, showing the possibility of baicalein as an antifibrogenetic drug for hepatic fibrosis. PMID- 10478574 TI - Comparison of vasopressin binding sites in human uterine and vascular smooth muscle cells. AB - Several studies indicate that oxytocin and vasopressin receptors in the human uterus are heterogeneous. We have investigated whether oxytocin and vasopressin bind to separate receptors or one class of receptors in human uterine smooth muscle cells. [3H]d(CH2)5Tyr(Me)AVP, the vasopressin V1A receptor selective radioligand, was used for comparison of vasopressin binding sites in human uterine and vascular smooth muscle cell membranes. Both membrane preparations exhibited one class of high-affinity binding sites with Kd values of 6.44 and 0.47 nM, Bmax values of 166 and 34.8 fmol/mg protein for uterine and vascular smooth muscle cells, respectively. In vascular preparations, the selective vasopressin V1A receptor antagonist, SR 49059 ((2S) 1-[(2R 3S)-(5-chloro-3-(2 chlorophenyl)- -(3.4-dimethoxybenzenesulfonyl)-3-hydroxy-2,3-dihydro-1H-indole-2- carbonyl]-pyrrolidine-2-carboxamide), showed high affinity with Ki value of 0.98 nM, confirming that these receptors belong to the vasopressin V1A receptor subtype. On the contrary, in uterine preparations, binding of [3H]d(CH2)5Tyr(Me)AVP was more effectively displaced by oxytocin and the oxytocin receptor selective antagonist, L-371257, (1-[1-[4-[ N-Acetyl-4-piperidinyl)oxy]2 methoxybenzoyl]piperidin-4-yl]- 4H-3,1-benzoxazin-2(1H)-one), than vasopressin and SR 49059, suggesting that binding may be due to cross-reaction with the oxytocin receptors. These results suggest that human uterine smooth muscle cells express only a high density of oxytocin receptors. PMID- 10478575 TI - Adrenaline influences the release of interleukin-6 from murine pituicytes: role of beta2-adrenoceptors. AB - In this study, we examined the effect of adrenaline and interleukin-1beta on interleukin-6 secretion from cultured murine neurohypophyseal cells. Cells were cultured from neurohypophyses of 3- to 5-week-old mice and experiments were performed after 13 days in culture. Interleukin-6 was measured in 24-h samples using a sandwich fluoroimmunoassay. Unstimulated cells released 19+/-3 fmol interleukin-6/neurohypophysis/24 h (mean +/- S.E.M., n = 42). Adrenaline and interleukin-1beta increased the release of interleukin-6 from the cells in a concentration-dependent manner. Incubation with adrenaline (10(-6) M) or interleukin-1beta (11 pM) induced maximal secretion of interleukin-6, resulting in a 2.2-fold and 19.8-fold increase, respectively (P<0.01). The action of adrenaline (10(-6) M) and interleukin-1beta (1.1 pM) was examined separately and together. The sum of the effect of the two compounds given alone was significantly less (P<0.05) than the effect when adrenaline and interleukin-1beta were given together. We examined the effect of the beta-adrenoceptor antagonist propranolol (3.4x10(-6) M), the beta2-adrenoceptor antagonist (+/-)-1-[2,3 (Dihydro-7-methyl-1H-inden-4-yl)oxy]-3-[(1-methyl-eth yl)amino]-2-butanol (ICI 118551) (10(-7) M) and the beta1-adrenoceptor antagonist atenolol (10(-7) M and 10(-6) M) on the adrenaline-stimulated release of interleukin-6. Propranolol and ICI 118551 completely blocked the action of adrenaline, whereas atenolol was inactive. It is concluded that the stimulatory effect of adrenaline is mediated via beta2-adrenoceptors. PMID- 10478576 TI - High prevalence of anti-alpha-crystallin antibodies in multiple sclerosis: correlation with severity and activity of disease. AB - INTRODUCTION: The presence of T-cell reactivity to alphaB-crystallin in patients with multiple sclerosis (MS) has suggested that this small molecular weight heat shock protein (Hsp) may be an autoantigen in MS. MATERIAL AND METHODS: We have tested the serum of patients with clinically definite MS (n=30), other inflammatory neurological disease (n=22), non-inflammatory neurological disease (n=42) and healthy individuals (n=23) for systemic humoral responses to bovine alphaB-crystallin, to the homologous chaperone protein, alphaA-crystallin, and to another small Hsp, Hsp 27. RESULTS: Sixty-three percent of MS patients exhibited immunoreactivity to alpha-crystallin and this was present in all 4 of 4 non ambulatory patients with MS. In contrast, serum concentrations in MS patients of antibodies to the small Hsp, Hsp27, and to myelin basic protein were negligible (P<0.001). Serum anti-alpha-crystallin immune responses were detected in significantly lower percentages of patients with other inflammatory neurological diseases (32%, P<0.025), and with non-inflammatory neurological diseases (12%, P<0.001). None of the healthy control individuals showed anti-alpha-crystallin reactivity. The concentration of anti-alpha-crystallin antibodies in patients with MS correlated with severe disease (P<0.05) and with active disease (P<0.025). CONCLUSION: Our observations support the notion that anti-alpha crystallin autoimmune responses may contribute to pathogenicity in MS and may represent a mechanism of how recurrent attacks of MS develop subsequent to an isolated demyelinating episode. PMID- 10478577 TI - A health-related quality of life questionnaire for multiple sclerosis patients. AB - OBJECTIVE: The construction of a brief, valid and reliable HRQoL questionnaire for use in multiple sclerosis patients based on generic and disease-specific HRQoL measures. MATERIAL AND METHODS: The Medical Outcomes Study 36-item Short Form Health Survey (SF-36), COOP/ WONCA Charts, and Disability & Impact Profile (DIP) were used in a longitudinal study in 162 patients with multiple sclerosis. RESULTS: Factor analyses identified 2 underlying dimensions of HRQoL, relating to "physical functioning" and "psychological functioning". Selection of the 3 highest loading reliable scales on each factor resulted in a final questionnaire consisting of 3 scales of the SF-36 and 3 scales of the DIP. In total 40 items were selected; completion time is about 10 min. CONCLUSION: The final questionnaire adequately measured 2 dimensions of HRQoL. The length of the questionnaire is acceptable for patients with MS in view of respondent burden. PMID- 10478578 TI - A multidimensional assessment of multiple sclerosis: relationships between disability domains. AB - OBJECTIVE: To test the feasibility of a multidimensional assessment based on both task-related and self-evaluation questionnaire scores in patients with multiple sclerosis (MS); ii) to correlate the results from selective measures with the severity of illness in terms of the Expanded Disability Status Scale (EDSS) score; iii) to assess the relationships between different domains of MS-related disability and handicap. PATIENTS AND METHODS: Eighty-three MS patients (M/F 31/52; age 43.26 +/- 10.9 years, range 21-72) underwent a standard clinical evaluation of motor abilities (by means of the Rivermead Mobility index, Timed Walking Test, Nine Hole Peg test and Hauser Ambulation Index) and cognitive performances (using Digit Symbol, Buschke-Fuld selective remind test, "FAS"-Word Fluency, Wisconsin Card Sorting test and Block design test). The Beck Depression inventory, MS Specific Fatigue Scale, Functional Assessment of MS and London Handicap Scale were applied to evaluate mood, fatigue, quality of life and handicap, respectively. Minimal Record of Disability measures - MRD (i.e. EDSS, Inability Status Scale and Environmental Status Scale) were also applied to test the criterion validity of the selected disability and handicap scales. The Kruskal-Wallis H-test for independent samples tested differences between subgroups with an increasing EDSS score (<3.5, 3.5-6.0, >6.0). The covariance and redundancy of measures included in the multidimensional assessment were evaluated through Factor Analysis. The Multiple Regression Analysis was used to detect the relative impact of either motor or cognitive disabilities and depression on handicap and quality of life. RESULTS: The multimodal assessment took 70 min on average to be performed, being well accepted by patients. Motor abilities worsened as the EDSS score rose, unlike cognitive performances which proved to be similarly impaired at different severity levels. Measures of fatigue and depression were not related to EDSS values. The chosen measures were assigned by Factor Analysis to 4 domains corresponding to motor performance, executive performance, cognitive abilities and quality of life, respectively. Regression analysis showed how handicap and depression independently affect quality of life. While the handicap score is mostly influenced by motor ability, as measured by the Rivermead Mobility Index, the depression score grows independently of any physical or cognitive disability and seems to be related to fatigue self assessment scores. CONCLUSIONS: A multidimensional approach to MS patient assessment allows a more detailed and sensitive evaluation of their disability profile and perceived difficulties, leading to a care programme tailored to the patient's needs. PMID- 10478580 TI - Epidemiological and clinical characteristics of myasthenia gravis in Belgrade, Yugoslavia (1983-1992). AB - This is the first epidemiological study of myasthenia gravis (MG) in the area of Belgrade. During the survey period (1983 1992), 124 incidental cases of MG were observed, producing an average annual incidence rate of 7.1 per million population (women, 8.3; men, 5.8). Age and sex specific incidence rates for females demonstrated a bimodal pattern, with the first peak in the age group between 20 and 40, and the second peak in the age group 70-80. The age-specific rates for males showed unimodal pattern, reaching a maximum in the age group between 60 and 80. There was a tendency of more frequent disease appearance in the urban as opposed to the suburban districts. On the prevalence day, December 31, 1992, the point prevalence rate was 121.5 per million (women, 142.5; men, 98.8). Only for incidental cases, the point prevalence rate was 77.1 (women, 83.2; men, 70.4). The average annual mortality rate was 0.47 per million (females, 0.52; males, 0.42), while cumulative lethality was 5.6 (women, 5.6; men, 5.7). Most frequently initial symptoms were ocular, occurring in 58% patients. Through the period of investigation ocular symptoms were generalized in 68%, most frequently in the first 2 years (62.5%). Thymoma was confirmed in 11.3% of patients. In this group there was equal presence of both sexes, older median age at onset, and more severe clinical course of MG. Associated autoimmune disease was found in 17 out of 124 incidental cases (13.7%). The most common were thyroid diseases (7.3%). Family history of MG was recorded in 2 cases belonging to 1 family (1.6%). PMID- 10478579 TI - Nocturnal subcutaneous apomorphine infusion in Parkinson's disease and restless legs syndrome. AB - OBJECTIVES: Nocturnal disabilities leading to fragmented sleep arising from parkinsonian off period related complications are common, under-reported and are difficult to treat. In this study, we evaluate the use of nocturnal continuous subcutaneous overnight apomorphine infusion in Parkinson's disease and restless legs syndrome. METHODS: Six parkinsonian patients and 2 patients with restless legs syndrome with nocturnal disabilities refractory to conventional oral therapy were assessed using a sleep diary while on standard treatment and during nocturnal apomorphine infusion. Three patients agreed to assessments during placebo infusion with normal saline. RESULTS: Apomorphine led to a dramatic reduction of nocturnal awakenings, nocturnal off periods, pain, dystonia and nocturia in parkinsonian patients. In patients with restless legs syndrome, apomorphine reduced nocturnal discomfort, reduced leg movements and improved pain and spasm scores significantly. Placebo infusion reproduced pain, nocturnal spasms and sleep disruption. CONCLUSION: This study suggests that overnight apomorphine infusion may be effective in overcoming refractory nocturnal disabilities in selected patients with Parkinson's disease and restless legs syndrome. PMID- 10478581 TI - An enzyme-linked immunosorbent assay for antibodies against acid soluble skeletal muscle antigen in myasthenia gravis. AB - An enzyme-linked immunosorbent assay (ELISA) system has been developed for measuring serum antibodies against citric-acid extract of human skeletal muscles. With this assay, 80% of myasthenia patients with thymoma gave positive results. No sera from patients with neurological diseases other than myasthenia gravis (MG) gave positive results. The result of this study indicates that the ELISA system is useful as an adjunct for the diagnosis of MG particularly in the patients associated with thymoma and that the ELISA method can also be used as a prognostic marker following thymectomy in patients with MG. PMID- 10478582 TI - Long-term course and outcome in AIDS patients with cerebral toxoplasmosis. AB - OBJECTIVES: We compared clinical long-term course and outcome in all AIDS patients admitted to our outpatient department from January, 1989 to June, 1998 with toxoplasma encephalitis (TE) as first AIDS-defining infection (n= 106) and in 106 patients with Pneumocystis carinii pneumonia (PCP) as first AIDS-defining disease. MATERIAL AND METHODS: The 2 groups were matched with respect to age, sex, risk group, degree of immunodeficiency as measured by CD4 cell counts and duration of HIV-1-positivity. TE was diagnosed radiologically and by response to treatment. RESULTS: Forty-three TE patients surviving the first TE symptoms > 14 months developed dementive symptoms, leucoencephalopathy in imaging procedures and died from dementia. In contrast only 5 patients surviving PCP for an equally long period showed dementive symptoms. CONCLUSION: Cerebral infections like toxoplasma encephalitis (TE) may negatively influence HIV-1-activity so far latent in the brain. PMID- 10478583 TI - Mechanically elicited nerve root discharge: mechanical irritation and waveform. AB - OBJECTIVE: Intraoperative monitoring is very important for protecting nerve roots during lumbar surgery for spinal degeneration. Our objective was to evaluate the correlation between the type of mechanical irritation and waveform by mechanically elicited EMGs during the surgery. METHOD: Mechanically elicited EMGs were recorded bilaterally on muscle groups innervated by the lumbar nerve roots in the area of surgery in 24 consecutive patients with lumbar spinal degenerative disease. RESULTS: It was confirmed that surgical irritation produced 3 types of waveforms as discharges from nerve roots; short, waning and continuous discharges. Each waveform was easily elicited mechanically and was sensitively related to the type and strength of mechanical stimulation. CONCLUSION: It was indicated that the mechanical irritation on the root could elicit the nerve root discharge with no post-operative nerve root deficit. It may be useful to monitor the mechanically elicited EMGs during the surgery for spinal degeneration. PMID- 10478584 TI - Identification of five spinocerebellar ataxia type 2 pedigrees in patients with autosomal dominant cerebellar ataxia in Taiwan. AB - OBJECTIVES: The autosomal dominant cerebellar ataxias (ADCAs) are a group of genetically diverse neurological conditions linked by progressive deterioration in balance and coordination. Spinocerebellar Ataxia Type 2 (SCA2) is one of the ADCAs and also belongs to a special group caused by the expansion of an unstable CAG repeat encoding a polyglutamine tract. We aimed to investigate the frequency of SCA2 mutation in the ataxia patients referred to the clinic. MATERIALS AND METHODS: We screened 58 families with inherent cerebellar ataxia and 57 normal individuals by the use of radioactive genomic polymerase chain reaction (PCR) method. A simple non-radioactive PCR for rapid detection of the expanded SCA2 alleles via agarose gel electrophoresis was also employed. RESULTS: Eight SCA2 affected patients and 1 at-risk individual in 5 unrelated SCA2 families were identified. The CAG repeats of normal alleles in the sample studied range in size from 16 to 30 repeat units, while those of SCA2 chromosomes are expanded to 34 to 49 repeat units. Our results also showed that unlike SCA 1 and SCA3/MJD, the size distribution of the normal alleles showed few polymorphisms, with the 22 repeat allele accounting for 90.1%. Homozygosity in normal individuals was 80.2%. No overlap in ataxin-2 allele size between normal and expanded chromosomes was observed. CONCLUSION: This is the first report of the SCA2 gene distributions in the population of Taiwan. The SCA2 mutation accounts for 8.6% of ADCA type I families referred to us, intermediate between SCA1(1.7%) and SCA3/MJD (24%) of the ADCA type I families in our collection. PMID- 10478585 TI - The genetic analysis of Turkish patients with Huntington's disease. AB - OBJECTIVE: Exploring the (CAG)n expansion within IT 15 gene in Turkish Huntington's disease (HD) patients and its relation to downstream (CCG)n repeat polymorphism to elucidate population specific haplotypic heterogeneity. METHODS: Twenty-seven patients with clinical diagnosis of HD from 19 families were sampled. The triplet repeats were evaluated by sizing the fluorescent PCR products on an ABI 310 capillary gel electrophoresis unit. RESULTS: The number of (CAG)n repeat expansions (range: 40-76, mean: 45.6+/-7) were inversely correlated with age of onset (r=-0.81, P<0.0001). The (CCG)n polymorphism in HD chromosomes was confined to (CCG)7 in all patients. In normal chromosomes CAG polymorphism was accumulated at a relatively higher range (mean: 19.3+/-2.9) together with the common occurrence of(CCG)7 and (CCG)10 alleles. CONCLUSION: The distribution range of the CAG and CCG repeat polymorphism in normal chromosomes and strong linkage disequilibrium between HD mutation and (CCG)7 haplotype provided a striking similarity to populations of western European descent. PMID- 10478586 TI - Autosomal dominant limb-girdle muscular dystrophy with ankle joint contracture. AB - We herein describe a Japanese family suffering from autosomal dominant limb girdle muscular dystrophy showing a later onset and a predilection for males. All patients developed a plantar flexion contracture of the ankles when no distal muscular weakness was evident. The serum creatine kinase activity was remarkably elevated to 55 times the normal value. These clinical features suggest a novel phenotype. PMID- 10478587 TI - On the prognosis of outcome after stroke. AB - OBJECTIVES: The study was aimed at improving the accuracy of prognosis for recovery of function in patients suffering a first stroke. MATERIALS AND METHODS: Two-hundred and forty-eight patients were enrolled. The mean interval since the stroke was 23 days. Patients entered a rehabilitation program lasting 60 days. The predictive value of 12 factors were analysed, namely motor, cognitive and sphincter subitems of Functional Independence Measure at admission (FIM-a), age, sex, education, body mass index (BMI), depression, neglect, aphasia, ideomotor and constructive apraxia. FIM score at discharge was the dependent variable. RESULTS: A multiple regression revealed that only age, cognitive and sphincter subitems of FIM-a, neglect and ideomotor apraxia were significantly associated with outcome. Moreover, these factors accounted for only 72% of the variance in outcome scores. A decision of unfavourable prognosis on the basis of a FIM-a value lower than 40 was incorrect in 2.8% of the patients in this study and in 8.2% of those having a FIM score lower than 40. CONCLUSIONS: The use of statistical methods to examine the outcome after stroke is useful for expressing probability on a group basis but is unsuitable for determining the prognosis of individual patients. Such data should not be used for fiscal management. A significant minority of patients presenting with a FIM lower than 40 can regain a useful measure of independence. The errors in prognosis based upon available methods, although small, have unacceptable effects in human terms if they lead to the clinical decisions which deny patients rehabilitation. All of the patients should therefore be admitted for rehabilitation after their first stroke. Severe comorbidity requires special attention. PMID- 10478588 TI - The hormone status of patients with multiple sclerosis. PMID- 10478589 TI - Otoneurological expert system for vertigo. AB - We have developed an otoneurological expert system (ONE) to aid the diagnostics of vertigo, to assist teaching and to implement a database for research. The ONE database is set to harvest data on patient history, signs and test results necessary for diagnostic work with vertiginous patients. A method based on pattern recognition was used in the reasoning process. Questions about symptoms, signs and test results are weighted and scored for each disease and the most likely disease is recognized from defined disease profiles. Missing information and uncertainties are solved with a method resembling fuzzy logic. ONE was validated by comparing diagnoses assessed by physicians with those provided by the system. It proved to be a valid decision-maker by solving 65% of the cases correctly, while the physicians' mean was 69%. To improve ONE further, a follow up should be implemented for the patients, since diagnosing sudden deafness and Meniere's disease during the first visit is often impossible. We aim to obtain new information on diseases involving vertigo by applying adaptive computer applications, such as genetic algorithms, to the reasoning process. PMID- 10478590 TI - Surgical treatment of vertigo by utriculostomy: an experimental study in sheep. AB - The objective of this study was to develop and test an endolabyrinthic microsurgical procedure for treatment of vertigo, the utriculostomy. This involves the application of local heat for obtaining a fistula in the membranous labyrinth, so as to establish communication between the endolymphatic and perilymphatic spaces at the utricle level. Before the procedure, an experimental model using quail eggs was built for pre-evaluation, and macroscopic and histological studies were performed in the temporal bones of three healthy sheep. Following this, the utriculostomy was performed through the oval window in 12 sheep. A microthermocautery was conceived by the first author and developed at Hospital de Clinicas de Porto Alegre. This equipment allows for control of temperature and duration of exposure to heat. Three months after the surgery, the animals were killed. A histological study of the temporal bones was performed to assess whether communication had been created between the endolymphatic and perilymphatic spaces, or whether a neomembrane had developed in the cauterized region. Histological sections of the vestibule of eight animals (three normal, five surgical) were analysed. All non-surgical cases presented a normal utricle wall. Three surgical cases (60%) presented a neomembrane. The absence of identifiable perforations in the utricle wall and the presence of neomembrane areas in 60% of the operated bones suggest that utriculostomy is a promising procedure for the treatment of Meniere's disease. PMID- 10478591 TI - Cogan's syndrome: clinical significance of antibodies against the inner ear and cornea. AB - The aim of this study was to evaluate the pathological significance of antibodies against cornea and inner ear tissue in the development of audiovestibular and ocular symptoms in patients with Cogan's syndrome (CS). We analysed the serum of 5 CS patients for binding of IgM and IgG to fresh cryosections of rat labyrinth (semicircular canals, ampulla, utricle, saccule) and cornea by indirect immunofluorescence (IF). The predominant pattern of anti-corneal IgM was staining of the superficial cell layer of the non-keratinizing squamous epithelium. IgM against cornea was found in 3 patients, all of whom had bilateral inflammatory eye signs at the start of the disease. However, IgM was also detected in the chronic stage of the disease when no clinical signs of eye involvement were apparent. The study includes the first follow-up examination of anti-corneal IgM and IgG antibodies during a complete episode of active CS. During the first episode of CS in 1 patient, anti-corneal IgM became detectable 1 week after the onset of interstitial keratitis and 3 weeks after the onset of audiovestibular symptoms. It increased over several weeks and then fell to very low levels. However, at no time was anti-corneal IgG found. In the course of follow-up examinations, the serum of 4 patients intermittently contained low titre IgG antibodies against inner ear labyrinthine tissue, but without any clear correlation with the active stages of CS. In addition, high-resolution MRI (HR MRI) of the inner ear was performed in the acute and chronic stages of CS to evaluate the activity of CS. In the acute stage, HR-MRI revealed abnormal MRI signals in the vestibule, semicircular canals, vestibular nerve, or cochlea. In the chronic stage, patients showed narrowing or occlusion of semicircular canals and the cochlea on the 3D-CISS images, but no high signal lesions (T1) and no enhancement. Antibodies against cornea or labyrinthine tissue were not consistently detected in CS and the level of organ-specific antibodies did not correlate with the activity of the disease. PMID- 10478592 TI - Measures of cochlear travelling wave delay in humans: I. Comparison of three techniques in subjects with normal hearing. AB - This study was designed to estimate and compare measures of cochlear travelling wave delay and travelling wave velocity in normally-hearing adults. Travelling wave delay and velocity measures were estimated in 23 normally-hearing adults using three different test techniques: i) derived auditory brainstem responses (ABR); ii) derived frequency-following responses (FFR); and iii) tone-burst evoked otoacoustic emissions (TBEOAE). Estimates from ABR and TBEOAE were comparable to each other in terms of both averaged group values and associated standard deviations. Furthermore, mean cochlear travelling wave velocity estimated from ABR and TBEOAE were comparable to published estimates. Average cochlear delays obtained using the derived FFR were significantly shorter than those obtained from the other two techniques, possibly due to the effect of cochlear microphonic contamination. Among the dependent variables investigated, measures of delay are to be preferred over velocity since the latter are based on uncertain values of cochlear distance. PMID- 10478593 TI - Expression of IsK protein mRNA in cultured rat strial marginal cells. AB - A cell culture system of marginal cells (MC) of the rat stria vascularis was established by the explant method. When grown on plastic dishes, cultured MC showed a polygonal "cobblestone-like" appearance. Dome formation, composed of several hundreds to thousands of cells, occurring after confluence suggested that vectorial transport of ion(s) with accompanying fluid developed in the cultured MC. Transmission electron microscopy demonstrated junctional complexes formed of tight junctions and desmosomes at the upper lateral membranes. The polymerase chain reaction (PCR) product, amplified with primers made from the cDNA reverse transcribed from cultured MC, yielded a distinct band compatible with the expected size of the PCR products amplified from cDNA of positive control groups containing IsK protein, indicating that cultured MC expressed the IsK protein mRNA. The results show that cultured MC can form large domes and express the most characteristic IsK protein, indicating that they maintain their vectorial electrolyte transport function and, possibly, the ability to secrete K+ in this condition. PMID- 10478594 TI - Changes in off-lesion endocochlear potential following localized lesion in the lateral wall. AB - Endocochlear potential (EP) was measured at various off-lesion sites after a small focal lesion was made in the lateral wall of the guinea pig cochlea. Lesions were produced by a photochemical reaction between systemically administered rose bengal and focused green light illumination. In 21 ears, continuous measurement for 30 min after onset of the reaction at turns apical or basal to the site of illumination revealed no significant changes in EP compared with the control value (p < 0.01). In another group of 43 ears, EP was measured at 3 days post-illumination. A significant decline was seen at every site located apical to the lesion (p < 0.001). Conversely, no significant change was measured at any site located basal to the lesion. These findings suggest that the decrease in EP assumes the form of a gradient from the lower to upper turns in the guinea pig cochlea. PMID- 10478595 TI - Can MRI replace a second look operation in cholesteatoma surgery? AB - The purpose of this paper is to investigate whether a magnetic resonance imaging (MRI) examination of the petrous bones could replace the conventional second look surgical procedure when a canal wall up tympanoplasty procedure has been performed during the first look surgery. Eighteen patients were examined with MRI prior to a second look surgical procedure. MRI was performed on a 1.5 T superconducting system. T2 weighted images before contrast and T1 weighted images after contrast (gadopentate dimeglumine) were obtained. Imaging results were compared with surgical findings. The poor radiosurgical correlation (50% and 61% after re-evaluation) suggests that, at the present time, MRI is not a valid alternative to a second look surgical intervention in the case of cholesteatoma treated by canal wall up tympanoplasty. In our study, MRI was not capable of differentiating small pearls of residual cholesteatoma from surrounding scar tissue. PMID- 10478596 TI - Relationship between severity of middle ear mucosal lesion and middle ear pneumatic space volume in patients with otitis media with effusion. AB - If we assume that the state of suppression of pneumatic cells is the result of suppression of pneumatic cell growth by inflammatory stimulation in the middle ear pneumatic space, it is possible to improve the state of suppression by performing sufficient treatment during the growth period of the pneumatic cells. We indwelt a tympanic membrane ventilation tube (hereinafter referred to as tube) for treatment of otitis media with effusion (OME) in child patients aged 3-13 years and investigated the following points: i) relationship between the severity of inflammation of the lamina propria of middle ear mucosal specimens (hereinafter referred to as lamina propria) collected at the time of tube indwelling and the degree of growth of the pneumatic space; and ii) changes in the pneumatic space associated with treatment by tube indwelling, which was studied by comparing the above-described mucosal severity with the pneumatic space area of 2 years after tube indwelling, and with increase in the pneumatic space volume measured periodically after tube indwelling. The results indicated that mastoid cell growth suppression is higher in patients with a higher degree of inflammatory changes in the lamina propria. In association with treatment by tube indwelling, effusion accumulated in the pneumatic space and mucosal swelling disappeared early after the treatment, or 2 months of tube indwelling. After that, in patients with severe mucosal lesion, a long time, 1.5-2 years, was found to be required for repneumatization accompanying regrowth of the temporal bone. We confirmed that the severity of inflammation of the lamina propria is deeply involved in the growth and repneumatization of the pneumatic cells. PMID- 10478597 TI - Epidemiology and aetiology of middle ear cholesteatoma. AB - A total of 500 patients with cholesteatoma diagnosed and operated during 1982-91 in the region of Tampere University Hospital and Mikkeli Central Hospital in Finland were analysed retrospectively. The mean annual incidence was 9.2 per 100,000 inhabitants (range 3.7-13.9) and during the study period the annual incidence decreased significantly. The incidence was higher among males under the age of 50 years. There was no accumulation of cholesteatoma diseases in lower social groups. The majority (72.4%) of cholesteatoma patients had suffered from otitis media episodes. Tympanostomy was carried out in 10.2% and adenoidectomy or adenotonsillectomy in 15.9% of all cholesteatoma ears prior to cholesteatoma surgery. Cholesteatoma behind an intact tympanic membrane with no history of otitis media was verified in 0.6% of patients and in cleft palate patients in 8%. In this study, 13.2% of patients had ear trauma or ear operation in anamnes. PMID- 10478598 TI - Evaluation of a cement incus replacement prosthesis in a temporal bone model. AB - In reconstruction of the ossicular chain for a damaged incus, it is important that the incus replacement prosthesis (IRP) length is ideal in order to provide optimal tension between the tympanic membrane or malleus and stapes head to achieve the best post-operative hearing result. Even though the length of commercially available IRPs can be adjusted, it still may be difficult to achieve clinically. We describe experiments in a human temporal bone model using dental cement as an IRP after removal of the incus. This cement IRP (CIRP) hardens in situ and becomes the length of the gap to be spanned so that tension should be ideal. Two different CIRPs were studied; one was a conventional rod-type CIRP connecting either the umbo or mid-malleus handle to the stapes head. The second was a Y-shaped CIRP (Y-CIRP), connecting two sites on the malleus to the stapes head. The wide Y-CIRP connected the malleus head and umbo to the stapes head, while the narrow Y-CIRP connected the malleus neck and mid-handle to the stapes head. The acoustic performance of these experimental CIRPs was studied using a laser Doppler vibrometer system in 12 fresh human temporal bones. The CIRP demonstrated better acoustic performance than conventional IRPs studied previously in the same model. While all the CIRPs showed similar function below 2.0 kHz, the narrow Y-CIRP appeared best above 3.0 kHz. A prosthesis of this type may have an acoustic advantage over conventional IRPs. PMID- 10478599 TI - An immunohistochemical study of inducible nitric oxide synthase in the rat middle ear, with reference to tympanosclerosis. AB - Tympanosclerosis and myringosclerosis are well-known sequelae after acute and chronic otitis media and are also often seen after treatment of secretory otitis media with ventilation tubes. They sometimes cause serious hearing disability. There is no successful treatment for these conditions. There might be factors triggering an immunological or autoimmune chain reaction, which leads to tympanosclerosis. Intervention with the aim of abolishing this type of response might be possible if an interruption of the chain reaction can be found. Nitric oxide is a radical molecule with the ability to kill pathogens and is produced by the enzyme nitric oxide synthase. Expression of inducible nitric oxide synthase (iNOS) was analysed immunohistochemically in a rat model of acute otitis media. In rats sacrificed at days 3 and 6 after inoculation. iNOS was also strongly expressed in the middle ear mucosa and in the tympanic membrane as well as in the inner ear. In control specimens as well as in infected ones. iNOS was expressed in the tissue of the external ear canal. In rats sacrificed at day 10 and after 3 months, iNOS was expressed at the same locations, although less frequently. These data indicate that iNOS expression is induced during acute otitis media and suggest that nitric oxide may be important in the host defence against ear infections. PMID- 10478600 TI - Localization of nontypeable Haemophilus influenzae endotoxin in the middle and inner ear during experimental otitis media. AB - Nontypeable Haemophilus influenzae (NTHi) lipooligosaccharide, the major component of H. influenzae endotoxin, was localized in the middle and inner ear subsequent to the resolution of experimental otitis media induced by this pathogen. A monoclonal antibody specific for the lipooligosaccharide of this strain was used to probe sections of middle and inner ear tissue and visualized by means of the avidin-biotin peroxidase complex technique. Sixteen to seventeen days post inoculation with either viable or formalin-inactivated NTHi, endotoxin could be localized in both the middle and inner ear at a time when the middle ear was culture negative. Our data demonstrate that endotoxin shed by NTHi during otitis media penetrates the inner ear and binds to both tissue components and inflammatory cells in both the middle and inner ear. PMID- 10478601 TI - Expression of histamine H1 and H2 receptors in human nasal mucosa. AB - mRNA encoding histamine H1 and H2 receptors were detected in the human nasal mucosa using reverse transcriptase-polymerase chain reaction. The possibility of local release in connection with specific target receptors suggests a role for histamine in the regulation of vascular tone, glandular secretion and epithelial functions. PMID- 10478602 TI - Effects of sinus surgery on asthma in aspirin triad patients. AB - The aspirin triad (nasal polyposis, asthma and sensitivity to aspirin) is a well recognized clinical entity, also known as aspirin-induced asthma (AIA). The sinusitis associated with AIA is often difficult to treat and aggravates the asthmatic symptoms. In order to evaluate the surgical treatment of sinusitis in AIA, 22 patients who underwent sinus surgery were studied. Twenty patients (90.9%) got any relief of their asthma symptoms from sinus surgery. Postoperative pulmonary function test 1 year after surgery showed statistically significant improvement over the preoperative one. Three of 5 patients (60%) who used systemic steroids were able to eliminate or reduce their dosages. Also, 8 of 17 patients (47.1%) who were using inhaled topical steroids reduced their dosages and statistical analysis showed a significant difference in the doses of topical steroid used before and after surgery. Subjective evaluation of 20 patients (90.9%) indicated that the sinus surgery was effective for their asthma condition; showing from mild to marked improvement. For AIA patients aggravated by sinus disease, we recommend sinus surgery to improve the quality of life. PMID- 10478603 TI - New objective and quantitative tests for gustatory sweating. AB - Two newly developed tests for gustatory sweating, providing both quantitative and topographic information, are presented. In both tests a paper stencil shaped to fit the complex anatomy of the parotid region is used. The blotting paper technique uses the difference in weight before and after gustatory stimulation to measure the amount of sweating. The iodine-sublimated paper histogram (ISPH) uses iodine sublimated office paper that changes colour when wet. The paper stencil is than digitized and a histogram algorithm applied to measure the area of sweating. A calibration of these tests with known and appropriate quantities of saline is presented. PMID- 10478604 TI - The artificial throat: a new method for standardization of in vitro experiments with tracheo-oesophageal voice prostheses. AB - After total laryngectomy, the voice can be restored successfully with a silicone tracheo-oesophageal voice prosthesis. Biofilm formation and subsequent deterioration of the silicone material of the prosthesis often limit the mean life of the device to an average of 3-5 months. Although device replacement can be considered an easy outpatient procedure, frequent replacements are inconvenient for the patient and may lead to malfunction of the tracheo oesophageal fistula. Further understanding of the process of development and inhibition of the colonization of these polymer surfaces requires several comprehensive clinical studies. However, in vivo research of the biomaterials of the voice prostheses is difficult and time consuming. In order to simulate the natural process of biofilm development under dynamic nutrient conditions, an artificial throat was developed. Biofilm developed on Groningen button voice prostheses in vitro could not be distinguished from that formed over several months in vivo. This method can be used as a standardized approach for studying functional and structural aspects of all commercially available indwelling and non-indwelling voice prostheses, including the Groningen button, Provox, Voice Master, Blom-Singer and others under various laboratory conditions. PMID- 10478605 TI - Deterioration of intraoral hole size identification after treatment of oral and pharyngeal cancer. AB - Thirty-one patients with a diagnosed malignant tumour of the oral cavity or pharynx were tested in hole size identification on four test occasions: before all treatment, after radiotherapy and 6 months and 1 year after surgical treatment. They were compared within groups as well as with a group of healthy reference individuals of the same age who underwent the same test procedure at a 2 months' interval. The oral group did not decline in hole size identification after radiotherapy, but did after surgery. The deterioration was persistent 1 year after surgery. The pharyngeal group did not change performance in hole size identification after radiotherapy, nor after surgery. It is obvious that surgery of the oral structures causes the deterioration. No correlation with damage to the lingual nerve could be registered. The oral cavity reacts as one unit, despite sensory input from two sides. The non-operated side does not compensate for the operated side. It is plausible that decreased oral sensory acuity, in recognizing hole size of the bolus, contributes to postoperative swallowing problems. PMID- 10478606 TI - Ki-67 positive fractions in benign and malignant thyroid tumours: application of flow cytometry. AB - We investigated the DNA ploidy pattern, cell cycle and the percentage of Ki-67 positive fractions in fresh surgical material from 17 benign and 33 malignant thyroid tumours using flow cytometry. DNA aneuploidy was not seen at all in benign tumours, but was seen in 3 out of 33 malignant tumours, suggesting that detection of DNA aneuploidy indicates malignancy, although the detection sensitivity was low. Regarding the cell cycle, there was no difference in the percentage of S-phase fractions (SPF) or G2 plus M phase fractions (G2M) between benign and malignant tumours. However, the percentage of Ki-67 positive fractions in malignant tumours (39.9 +/- 3.9%) was significantly higher than that in benign tumours (9.4 +/- 2.1%), indicating that malignant thyroid tumours contained a large population of G phase cells. When a cut-off value of 20%, was used for Ki 67 positive fractions, sensitivity was 82%, specificity was 88% and accuracy was 84% for the diagnosis of malignant tumours. Although this study was carried out on surgically derived materials, it is possible that flow cytometric analysis of fine needle aspiration-derived materials may have a place in preoperative histopathological assessment of thyroid tumours. PMID- 10478607 TI - Speech articulation after subtotal glossectomy and reconstruction with a myocutaneous flap. AB - Speech samples of 9 subjects (8 males, 1 female) were recorded before and 0.5-2 years after a partial glossectomy and reconstruction with a pectoralis major myocutaneous flap. A reading sample, a list of meaningful and nonsense words, and a list of sustained vowels were recorded. The speech samples were evaluated by pairs of naive listeners and using acoustic analysis of the vowel production. Each pair listened to the recordings of only one patient. Inter-rater agreement was satisfactory. The general impression of the speech outcome varied from normal to moderately impaired. The perceptually estimated impairments of speech articulation in the after/before comparisons were statistically significant. Only the first formant of the vowel /i/ (rise) and the second formant of the vowel /a/ (drop) changed significantly at the group level. There was a negative correlation (r = -0.79) between the extent of tongue resection and the drop of the second formant of the vowel /a/. The perceptual variables showed a relationship (r = 0.74-0.82) with the changes in the level of the second formant of the vowel /i/. The relationship that emerged between the perceptual estimates and the objective acoustic parameters suggests that it will be possible to develop clinically relevant test batteries for articulatory quality analysis. PMID- 10478608 TI - New perspectives for asthma treatment: anti-leukotriene drugs. AB - Leukotrienes are synthesized by different cells, including eosinophils, neutrophils, basophils, lymphocytes, macrophages, and mast cells. Cysteinyl leukotrienes (LTC4, LTD4, and LTE4) are the most important leukotrienes in the pathogenesis of asthma. Pharmacologically, there are two different ways of blocking the action of leukotrienes: inhibiting their production by blocking 5 lipoxygenase or its activating protein, 5-lipoxygenase-activating protein (FLAP), (inhibitors) or by blocking their receptors (antagonists). The available antagonists are, for the moment, directed against the one receptor demonstrated to play a role in asthma symptoms, CysLT1, and they act in a competitive way. The only marketed 5-lipoxygenase inhibitor is zileuton (Zyflo). CysLT1 antagonists, currently on the pharmaceutical market in some countries, are zafirlukast (Accolate), pranlukast (Ultair, Onon), and montelukast (Singulair). Undoubtedly, drugs acting on leukotrienes constitute a new pharmacologic class in the therapeutic armamentarium for the management of asthma. From the pediatric point of view, montelukast is currently the most interesting drug of the group to date because of published trials in patients as young as 6 yr of age. At present, zafirlukast is only approved for use in patients 12 yr of age and older, although we understand that applications are likely to extend the age range into childhood shortly. However, more experience is necessary to establish a definite place for both leukotrienes in the step-by-step asthma treatment. New comparative studies (with sodium cromoglycate and inhaled steroids), which will probably be published in the near future, as well as studies on the use of montelukast in the treatment of children under 6 yr of age will add crucial information to our knowledge, and help to identify an appropriate use in the therapeutic algorithm. Montelukast will not be a substitute for inhaled corticosteroids or beta-agonists, although it may act as a 'sparing drug' (which might help tapering of steroids in some instances). Its role in exercise-induced asthma seems promising. Although its more widespread use could highlight low-frequency adverse effects, its apparent excellent tolerability is an additional advantage for the drug. Patients' preference for a twice-daily dosage over the inhaled medication, resulting in a better compliance, is relatively well established, at least for zafirlukast in adults, and these findings can probably be extended to montelukast, which only requires once-daily dosing in children. PMID- 10478609 TI - Perinatal risk factors for recurrent wheeze in early life. AB - The possible value of tidal flow volume (TFV) loops measured at birth in relation to the risk of developing recurrent or persistent bronchial obstruction within two years of life was assessed. TFV loops were measured at a mean age of 2.7 days in 802 neonates enrolled in the 'Environment and Childhood Asthma' (ECA) study in Oslo. Of these, 77 children developed recurrent or persistent bronchial obstruction (cases) and were included in a nested case-control study within the ECA study; 88 controls (the child born closest in time to the case), with no history of bronchial obstruction in the first two years of life, were also included. Information on socio-economic factors, parental atopic diseases and parental smoking habits during the pregnancy was collected from a questionnaire completed by the parents in the maternity ward, and cord blood IgE (CB-IgE) was determined as part of routine sampling in the delivery ward. Mean tPTEF/tE (time to reach peak flow to total expiratory time) was slightly lower in cases (0.31; 95% CI 0.28-0.34) than in controls (0.33; 0.31-0.35) (difference not significant), whereas geometric mean CB-IgE was significantly higher among cases (0.39; 0.30-0.52) than controls (0.27; 0.23-0.33). No significant differences between cases and controls were found for respiratory rate, peak tidal expiratory flow or expiratory volume. However, the odds ratio for developing recurrent or persistent bronchial obstruction was 3.5 (1.1-11.6) if tPTEF/tE was < 0.20 and 4.1 (1.1-14.5) with maternal daily smoking during the pregnancy, after adjusting for age, weight, sex, CB-IgE, parental atopy, maternal education and family income. The TFV parameter tpTEF/tE < 0.20 measured within the first week of life as well as maternal daily smoking during pregnancy are significant, independent risk factors for developing recurrent or persistent bronchial obstruction within the first two years of life. PMID- 10478611 TI - Age at adoption, ethnicity and atopic disorder: a study of internationally adopted young men in Sweden. AB - Epidemiological and laboratory studies have implied that the environment during early childhood is important for the risk of developing atopic disorders. In this study we analyzed the prevalence of asthma, hayfever and eczema among 1901 internationally adopted young men at the military-induction medical examination in relation to indicators of their early childhood environment. The adopted young men who came to Sweden before 2 years of age suffered from asthma, hayfever and eczema significantly more often than those who came to Sweden between 2 and 6 years of age; the risk ratios (RR) were 1.6, 2.5 and 2.1, respectively. The young men who were born in the Far East were identified as being particularly susceptible to the development of hayfever and eczema, with RRs of 1.3 and 1.7. This study demonstrates that the environment during the first 6 years of life has a profound influence on the risk of suffering from atopic disorders as young adults. PMID- 10478610 TI - The relative importance of socio-economic status, parental smoking and air pollution (SO2) on asthma symptoms, spirometry and bronchodilator response in 11 year-old children. AB - The aim of this study was to evaluate the relative contribution of several risk factors to the prevalence of allergic respiratory symptoms, and the positivity of the bronchodilator test with fenoterol, and to establish the relative importance of these factors on the variability of FVC, FEV1, PEF, MEF25, MEF50 and MEF75. A total of 340 11-year-old children attending school in polluted and non-polluted areas of the city of Cartagena, Spain, were studied. The polluted area had had an annual mean of 75 microg/m3 of SO2 over the last 10 years and the non-polluted area had < 20 microg/m3 during this period. A questionnaire about allergic respiratory symptoms was completed by the parents. Specific questions about parental smoking habits and socio-economic level were included. Each child's performance in spirometry before and after administration of 0.2 mg of inhaled fenoterol was evaluated. The only significant predictive variables in the logistic regression (for suffering any symptom or a positive bronchodilator response) were male sex for nasal symptoms (RR 1.37; p = 0.04) and housing near heavy traffic for eye symptoms (RR 1.45; p = 0.01). Living in the polluted area reduced the risk of a positive bronchodilator response (RR 0.61; p = 0.004). Maternal smoking, even though not statistically significant, tended to increased the risk of suffering any symptom (RR 1.26; p = 0.07) or of having a positive bronchodilator response (RR 1.23; p = 0.1). None of the risk factors studied was of significant importance in explaining the variability of spirometry results. Although none of the risk factors were specifically determinant to the symptom questions, bronchodilator test or spirometric measurements, having a mother who smokes seems more important than living in a polluted area if statistically non significant trends are considered. PMID- 10478612 TI - Effect of pre-immunization by killed Mycobacterium bovis and vaccae on immunoglobulin E response in ovalbumin-sensitized newborn mice. AB - A recently advanced hypothesis suggests that decreased exposure to T-helper (Th) 1-inducing agents causes Th2-biased differentiation in response to concomitant allergens. We therefore examined the effect of pre-immunization with killed Mycobacterium bovis and killed M. vaccae which are known to be very potent inducers of Thl immune response, on serum IgE response in ovalbumin (OVA) sensitized newborn mice. Eighty-four newborn Balb/c mice were divided into four groups and were immunized intraperitoneally 24 h after birth with 50 microl of 5 x 10(4) colony-forming units (c.f.u.) of killed M. bovis in group I (M. bovis group, n = 19), with 25 microl of 2.5 x 10(8) c.f.u. of killed M. vaccae plus 25 microl of 5 x 10(4) c.f.u. of killed M. bovis in group II (M. vaccae + M. bovis group, n = 28) and with 50 microl of only phosphate-buffered saline (PBS) in group III (no mycobacterial immunization, n = 18). No injection was applied to mice in group IV (control group, n = 19). Starting from 8 weeks of age, all mice except the control group were sensitized with 0.5 ml of 20 mg/ml OVA administered intraperitoneally 7 times every other day. Thirty days after the final injection, all animals except those in the control group were challenged with an aerosol of 2 mg/ml OVA. Forty-eight hours later, blood was collected from all mice for determination of serum IgE levels. A statistically significant difference was observed in the serum total IgE levels between groups III and IV (p = 0.0099), indicating that the mice were successfully sensitized with OVA. Serum total IgE values of the female mice in M. bovis group were found to be significantly lower than group III (p = 0.009), while no difference was observed in males. Serum total IgE levels of the M. vaccae + M. bovis group were found to be significantly lower than group III both in male and female mice (p < 0.0001 and p = 0.0001, respectively). Female values were even lower than controls (p = 0.0092). Pre immunization in the newborn period with killed M. bovis alone or in addition to M. vaccae may potentially be helpful in down-regulating an IgE response. PMID- 10478613 TI - Development of immunoglobulin G subclass antibodies to ovalbumin, birch and cat during the first eight years of life in atopic and non-atopic children. AB - Immune responses to allergens in young children include both Th1- and Th2-like immunity, which may regulate the secretion of immunoglobulin (Ig) G subclass antibodies differently. The time, route and level of exposure to an allergen may be decisive with regard to whether sensitization or tolerance will ensue. To study this, we investigated the development of IgG subclass antibodies to food and inhalant allergens during childhood. The study group comprised a cohort of 96 children participating in a prospective study. IgG subclass antibodies to ovalbumin, Bet v 1 and cat dander were analyzed at birth, 6 and 18 months and 8 years by ELISA. IgG1 and IgG3 subclass antibodies to ovalbumin peaked at 18 months and then declined up to 8 years of age, whereas antibodies to the inhalant perennial allergen cat, but not the inhalant seasonal allergen birch, increased with age. Exposure to cat and birch tended to be associated with high antibody levels to those allergens, whereas antibody levels to ovalbumin were not related to exposure to egg. The presence of positive skin prick tests and circulating IgE antibodies correlated with high levels of IgG subclass antibody responses to the allergens. Atopic symptoms were associated with high levels of IgG subclass, particularly IgG4, antibodies to the allergens. The difference in antibody levels between atopic and non-atopic children was most marked at 6 months for ovalbumin. For the seasonal inhalant allergen birch, the difference was apparent from 18 months, whereas a difference in antibody levels to the perennial inhalant allergen cat was already present at 6 months. In conclusion, IgG subclass antibodies to food allergens peak in early infancy and are then down-regulated, whereas antibodies to the inhalant perennial allergen cat, but not the inhalant seasonal allergen birch, increase with age. Atopy is associated with high levels of IgG subclass, particularly IgG4, antibodies to allergens, supporting a deviation of the immune system towards Th2-like responses in atopic children. PMID- 10478614 TI - Prevalence of parentally perceived adverse reactions to food in young children. AB - A substantial number of parents perceive that their children have adverse reactions to food, but it is well documented that objective assessments agree with only one-quarter to one-half of parentally reported reactions. In order to prevent wrong diagnoses and curtail unnecessary or inadequate diets, primary health care providers need to deal with the parental perception of adverse reactions to food. A description of the prevalence and pattern of parentally perceived adverse reactions to food in children is needed to meet this challenge. The aim of the present study was to estimate the prevalence, incidence and cumulative incidences of parentally perceived adverse reactions to food in children younger than 2 years of age, and to study the duration of the reactions. A population-based cohort of 3623 children born in Norway was followed from birth until the age of two. At 6-month intervals, the parents completed questionnaires regarding the occurrence and type of any reaction to food. Information was available on the outcome measure at all age points for 77.4% of the families and these were used in the analyses; 3.8% of the cohort were entirely lost to follow up. The cumulative incidence of adverse reactions to food was 35% by age two. Fruits, milk and vegetables accounted for nearly two-thirds of all reported reactions. Milk was the single food item most commonly incriminated, the cumulative incidence being 11.6%. The cumulative incidences of reported reactions to fruits and vegetables were 20.4% and 7.3%, respectively, with citrus fruits, strawberry and tomatoes as the most common food items in these groups. The cumulative incidences were less for food reactions associated with eggs (4.4%), fish (3%), nuts (2.1%) and cereals (1.4%). The duration of the reactions was short - approximately two-thirds of the reactions were not reported again 6 months later. However, the probability of remission depended on the food item concerned, the age at onset of reactions, and whether the reaction had been reported previously or not. Adverse reactions to food are reported by the parents of one-third of children in Norway before the age of two. The most striking feature of this study is the short duration of the food reactions, as approximately two-thirds of the reactions are not reported again 6 months later. Nevertheless, the high frequency of reactions attributable to milk is of concern. Milk is an important part of the Norwegian diet for children, and if removed from the diet its nutritional value is not easily replaced. Further studies are needed to assess the degree to which parents alter the diet of their children based upon perceived reactions to food. PMID- 10478615 TI - Evaluation of the risk of anaphylactic reactions by wasp venom-extract challenges in children. AB - Diagnostic sting challenges have been shown to provide information on the risk of further anaphylactic reactions to bee stings. We present a follow-up study in wasp venom-hypersensitive children after diagnostic venom extract challenges to analyze their risk of further anaphylactic reactions. Responses were obtained from 104 patients with wasp venom hypersensitivity out of 115 former patients. Only one of the 104 patients showed more than a severe local reaction to the sting challenge irrespective of the performance of a single or sequential challenge; therefore, only one patient received venom immunotherapy. The performance of a diagnostic sting challenge with wasp venom extract in children had a high negative predictive value of 94.6% for the risk of further systemic reactions. This was shown by analysis of later field stings, since 37 children experienced further field stings and only two of these children(5.4%) developed a mild systemic reaction (urticaria) equal to or less severe than the index sting. The value of the venom extract challenge can be interpreted in two ways: either it is less sensitive than a native sting challenge since the rate of systemic reactions to the challenge was very low, or the prognosis of wasp venom hypersensitivity in children is extremely favorable. Since the latter hypothesis is supported by the low incidence of systemic field sting reactions, we postulate that venom immunotherapy is necessary only in a minority of children with wasp venom hypersensitivity with an index sting reaction of Mueller grade I or II. However, the value of venom extract challenges as a general diagnostic instrument in children with Mueller I and II reactions due to wasp venom hypersensitivity may be questioned. It may have a place as a safe procedure in demonstrating to parents and physicians the often self-limiting natural course in most of these children. PMID- 10478616 TI - Comparison between serial skin-prick tests and specific serum immunoglobulin E to mite allergens. AB - Sensitization to dust mite allergens can be determined by means of a skin-prick test (SPT) or by measurement of specific IgE antibodies in serum (sIgE). In our study, concordance of the results of both methods was analyzed on the basis of reproducible SPT results. Three consecutive SPTs were performed on 138 school children (age 6-8 years) at one-year intervals. SIgE was determined at the end of a two-year observation period. Seven common inhalant allergens (Dpt, Df, birch pollens, hazel pollens, grass pollens and cat and dog dander) were analyzed. The majority of subjects with positive SPT reactions to the respective allergen also showed sIgE (Dpt: 82/86; Df: 53/53; cat dander: 31/32; dog dander: 6/9; birch pollens: 29/31; hazel pollens: 22/22; grass pollens: 37/37). A significant correlation between the SPT [weal diameter (P1) or allergen/ histamine ratio (P2)] and sIgE was found for Dpt (P1 = 0.004/ P2 = 0.016), birch pollens (P1 = 0.002/P2 = 0.0001) and grass pollens (P1 = 0.0005/P2 = 0.0001). There was also a significant correlation between sIgE to Dpt and to either Der p 1 (p = 0.0001) or Der p 2 (p = 0.0001), as well as between sIgE of both major allergens (p = 0.0001). In the analysis of co-sensitization of Dpt and Df, most subjects sensitized to Dpt were also sensitized to Df (57/91). Children with sIgE to Dpt (n = 87) usually showed sIgE to Df(n = 83). In this study, SPT and sIgE results are concordant and appear equivalent when using reproducible SPTs. Therefore, in the case of a positive Dpt result, additional testing for sensitization to Df can be regarded as redundant when Dpt and Df are the major contributors to the allergen content of house dust. PMID- 10478617 TI - Anti-Streptococcus mutans antibodies in saliva of children with different degrees of dental caries. AB - The aim of this study was to evaluate the relationship between the secretory immune system and dental caries. Forty-nine 3-5-year-old children with primary dentition were classified into three groups according to their caries indices: no caries (group I), one or two surfaces with caries lesions (group II) and rampant caries (group III). Lower numbers of mutans streptococci were found in group I in relation to groups II and III. Secretory IgA and anti-S. mutans IgA, IgM and IgG antibody levels were not significantly different among the groups. Western blotting analysis showed that some S. mutans proteins, including the 39, 59, 97 and 150 kDa molecular mass bands, were recognized by almost all the saliva samples. Antibodies against the 185 kDa band, known as antigen I/II, were present in all adults' saliva and in only one child in group III. The absence of antibodies to the 185 kDa band in children's saliva suggest a specific immunologic immaturity. Further prospective studies will be necessary to establish the possible effect of reactivity to this antigen on the S. mutans colonization in this age group. PMID- 10478618 TI - Glenohumeral instability: evaluation using MR arthrography of the shoulder. AB - In the setting of glenohumeral instability or when internal derangement of the shoulder joint is suspected, MR arthrography has been demonstrated to be an accurate diagnostic imaging technique. Knowledge of the complex anatomy of the shoulder and its variations is essential in order to maximize diagnostic accuracy. PMID- 10478619 TI - Magnetic resonance imaging features of allografts. AB - OBJECTIVE: To investigate the magnetic resonance imaging (MRI) features of allografts at various time intervals after surgery in patients with osteoarticular allografts. DESIGN AND PATIENTS: Sixteen patients who were treated with osteoarticular allografts and who were followed over time with MRI studies as part of their long-term follow-up were retrospectively selected for this study. T1-weighted images were obtained both before and after gadolinium administration along with T2-weighted images. All images were reviewed by an experienced musculoskeletal radiologist, with two other experienced radiologists used for consultation. Imaging studies were organized into three groups for ease of discussion: early postoperative period (2 days to 2 months), intermediate postoperative period (3 months to 2 years), and late postoperative period (greater than 2 years). RESULTS: In the early postoperative period, no gadolinium enhancement of the allograft was visible in any of the MR images. A linear, thin layer of periosteal and endosteal tissue enhancement along the margin of the allograft was visible in images obtained at 3-4 months. This enhancement appeared gradually to increase in images from later periods, and appears to have stabilized in the images obtained approximately 2-3 years after allograft placement. The endosteal enhancement diminished after several years, with examinations conducted between 6 and 8 years following surgery showing minimal endosteal enhancement. However, focal enhancement was noted adjacent to areas of pressure erosion or degenerative cysts. All the cases showed inhomogeneity in the marrow signal (scattered low signal foci on T1 with corresponding bright signal on T2), and a diffuse, inhomogeneous marrow enhancement later on. CONCLUSION: We have characterized the basic MRI features of osteoarticular allografts in 16 patients who underwent imaging studies at various time points as part of routine follow-up. We believe that the endosteal and periosteal enhancement observed on MRI during the first few months to 2 years following surgery represents vascular ingrowth and early skeletal repair. The zone of periosteal enhancement could also include the new bone laid on the surface of the allograft through which the soft tissues bind to the cortex. The exact reason for the inhomogeneity in the marrow signal, and the diffuse, inhomogeneous marrow enhancement is not clear. This may represent saponified and/or necrotic marrow fat interspersed with the fibrovascular tissue. The features noted here should provide radiologists with useful information regarding imaging characteristics they can expect to see in other allograft replacement patients. PMID- 10478620 TI - Fibrogenesis imperfecta ossium: imaging correlation in three new patients. AB - OBJECTIVE: Fibrogenesis imperfecta ossium is an extremely rare disorder that can easily be misdiagnosed. We retrospectively reviewed the clinical and imaging data of three confirmed cases of fibrogenesis imperfecta. DESIGN AND PATIENTS: The patients consisted of two men and one woman, ranging in age from 40 to 53 years. Radiography was performed in all the patients. One patient had a 3-year follow-up of the thoracolumbar spine with conventional radiography and thoracolumbar magnetic resonance (MR) imaging. Open biopsy was performed in all cases, confirming the diagnosis of fibrogenesis imperfecta ossium. RESULTS: All our cases demonstrated "fishnet" trabecular pattern by conventional radiographs, and a pelvis radiograph of one patient showed an equivocal sclerosis pattern. Multiple fractures were noted in two patients. A pseudoexostosis was present in the ilium in one patient. Thoracolumbar MR imaging demonstrated diffuse low signal intensity within the medullary space on both T1-weighed and T2-weighted images, except for a region of increased signal intensity in the L1 and L2 vertebral bodies on T2-weighed images due to edema from acute collapse. CONCLUSIONS: Although uncommon, fibrogenesis imperfecta ossium should be considered in a previously healthy patient with a combination of progressive bone pain, unexplained fractures, a radiologic pattern of fishnet osteopenia and MR imaging of low signal intensity bone marrow on both T1-weighted and T2-weighted images. PMID- 10478621 TI - The radiological findings in chronic expanding hematoma. AB - OBJECTIVE: To identify the characteristic MRI findings of chronic expanding hematoma correlated with the pathology. DESIGN AND PATIENTS: Three patients who had a chronic expanding hematoma involving the musculoskeletal system were reviewed retrospectively. RESULTS AND CONCLUSION: Huge soft tissue masses suggestive of malignancy with destruction of the bony structure were revealed on radiography and computed tomography. MRI showed the masses to exhibit heterogeneous signal intensity on both T1 and T2-weighted images with a peripheral rim of low signal intensity, reflecting the central zones of fluid collection due to fresh and altered blood with a wall of collagenous fibrous tissue. These MRI findings were seen in all three patients and are considered to be characteristic; they assist in differentiation from neoplasm in consideration of the history of trauma or surgery. PMID- 10478622 TI - Synovial desmoplastic fibroblastoma of hip joint with bone erosion. AB - A 78-year-old woman presented with pain at the left hip. Initial radiographs showed minimal age-related changes. Over the next 2 years she developed large erosions in the left femoral neck and proceeded to hip replacement. Histological examination showed bland spindle cells in a loose and hyalinised collagenous stroma considered to represent a desmoplastic fibroblastoma (collagenous fibroma). This is the first reported case of synovial desmoplastic fibroblastoma. PMID- 10478623 TI - Fibroma of tendon sheath of the infrapatellar fat pad. AB - We report on a 13-year-old boy who was found to have a fibroma of the tendon sheath associated with the patellar tendon and within Hoffa's fat pad of the knee. This benign tumor has never been described in this location previously. The MRI characteristics are correlated with the histologic findings. PMID- 10478624 TI - Deep soft tissue leiomyoma of the thigh. AB - A case of ossified leiomyoma of the deep soft tissues of the left thigh is presented. The radiographic appearance suggested a low-grade chondrosarcoma. MRI of the lesion showed signal characteristics similar to muscle on both T1- and T2 weighted spin echo sequences with linear areas of high signal intensity on T1 weighted images consistent with medullary fat in metaplastic bone. Histopathological examination of the resected specimen revealed a benign ossified soft tissue leiomyoma. PMID- 10478625 TI - Multifocal osteosarcoma as second tumor after childhood retinoblastoma. AB - We present a case of multifocal osteosarcoma (MFOS) arising 11.5 years after successful treatment of bilateral retinoblastoma. The clinical, imaging and pathological findings at onset, after therapy, and during follow-up are described. Fluorescent in situ hybridization did not reveal a deletion of the RB 1 retinoblastoma gene, although the presence of an inactivating mutation invisible to this method cannot be ruled out. The MFOS may have been a second multifocal tumor associated with the original retinoblastoma or a post irradiation sarcoma with extensive metastases. PMID- 10478626 TI - Influence of the non-competitive NMDA receptor antagonist MK-801 on 2-deoxy-D glucose-induced hyperphagia in rats. AB - The effects of the glutamate N-methyl-D-aspartate (NMDA) receptor antagonist (5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclo-hepten-5, 10-imine (MK 801) on 2-deoxy-D-glucose-induced hyperphagia were investigated in rats. MK-801 significantly increased 2-deoxy-D-glucose-elicited eating. The facilitating effects of MK-801 on 2-deoxy-D-glucose-elicited feeding were not affected by coadministration of a nitric oxide (NO) precursor, L-arginine. Because NO synthase inhibitors inhibit 2-deoxy-D-glucose-induced hyperphagia and activation of the NMDA receptor leads to NO formation, our results suggest that blockade of the NMDA receptor increases 2-deoxy-D-glucose-induced hyperphagia, which is unrelated to inhibition of NO, and that NMDA receptors may play a role in satiety. PMID- 10478627 TI - The neuroprotective glycine receptor antagonist GV150526 does not produce neuronal vacuolization or cognitive deficits in rats. AB - The neuroprotective activity of the novel glycine receptor antagonist (E) 3[(phenylcarbamoil)ethenil]-4,6-dichloroindole-2-c arboxylic acid sodium salt) (GV150526) was recently reported in a model of focal ischemia in the rat. Here it was investigated whether GV150526 treatment results in any of the adverse side effects commonly detected after injection of NMDA (N-methyl-D-aspartate) receptor antagonists. First, it was found that neuronal vacuolization in the posterior cingulate/retrosplenial area of the cortex was not induced by GV150526 (200 mg/kg, i.v.), but was evident after injection of the NMDA receptor antagonist dizocilpine (MK801) (1 mg/kg, s.c.). In a second set of experiments, the effects of GV150526 were examined on perforant path-dentate gyrus long-term potentiation in rats. GV150526 (3 mg/kg, i.v.) injected 30 min or 150 min prior to tetanization did not block potentiation of the e.p.s.p. slope and population spike amplitude. In contrast, in animals treated with MK801 (1 mg/kg, i.p.) 150 min before tetanization there was a clear block of long-term potentiation of the e.p.s.p. slope and population spike amplitude. The effects of GV150526 were also examined in the Morris Water Maze. Rats injected with GV150526 (10 mg/kg or 60 mg/kg, p.o.) did not show any impairment in learning when compared to control. MK801 (0.08 mg/kg, i.p.), on the other hand, significantly affected the ability to locate the escape platform in the Water Maze. These findings show that GV150526 is devoid of adverse side effects even at doses well above those producing a neuroprotective effect. This, drug has therapeutic potential with a much greater margin of safety than NMDA channel blockers or competitive NMDA receptor antagonists. PMID- 10478628 TI - Do alpha2-adrenoceptors play an integral role in the antinociceptive mechanism of action of antidepressant compounds? AB - Antidepressants are analgesic in the absence or presence of depression. The underlying mechanisms probably involve a complex interplay between several neurotransmitter systems and neuroreceptors. Alpha-adrenoceptors play an important role in pain processing and alpha2-adrenoceptor agonists have been used in clinical pain management so we have investigated whether alpha-adrenoceptor sub-types mediate the antinociceptive activity of antidepressants. Thus, the abdominal constriction assay in mice was used to examine the antinociceptive responses of a diverse range of antidepressants following alpha1- or alpha2 adrenoceptor antagonism. The antidepressants or monoamine reuptake inhibitors included the serotonin selective reuptake inhibitor paroxetine, the serotonin noradrenaline reuptake inhibitor sibutramine, the resolved (+)- and (-) enantiomers of the noradrenaline reuptake inhibitor oxaprotiline, plus the tricyclics amitriptyline and dothiepin. All these compounds have been previously shown to be antinociceptive in this paradigm. The respective alpha1- and alpha2 adrenoceptor antagonists prazosin and RX821002 ([2-(2-methoxy-1,-4-benzodioxan-2 yl)-2-imidazoline]) did not produce antinociception though at 1.0 mg kg(-1); s.c., RX821002 but not prazosin blocked clonidine antinociception. The antinociceptive activity produced by sub-maximal doses of amitriptyline, dothiepin, sibutramine, paroxetine, (+)- and (-)-oxaprotiline were all blocked by RX821002 but not by prazosin. Additionally, both morphine and aspirin antinociception was resistant to alpha1- and alpha2-adrenoceptor antagonism. Thus, alpha2- rather than alpha1-adrenoceptors may play an integral role in antidepressant antinociception irrespective of the propensity for inhibiting reuptake of not only noradrenaline but also serotonin. It is probable, however, that other differing pharmacological properties of some antidepressants, such as opioid-like activity, may complicate any empirical correlation between monoamine uptake and analgesia. PMID- 10478629 TI - Effects of terfenadine, astemizole and epinastine on electrocardiogram in conscious cynomolgus monkeys. AB - We examined the effects of non-sedative histamine H1 receptor antagonists on the electrocardiogram (ECG) in conscious cynomolgus monkeys. Terfenadine (3 mg kg(-1) h(-1), i.v.) and astemizole (0.3 and 1 mg kg(-1) h(-1), i.v.) caused significant time-dependent increases in the QT interval and QTc Bazett (QTc). However, normal ECG forms were found during a 60-min infusion of epinastine (3 mg kg(-1) h(-1) i.v.). A higher dose of epinastine (10 mg kg(-1) h(-1), i.v.) increased the QTc and PR interval only 5 min after the start of the infusion. The minimum plasma concentrations of terfenadine, astemizole and epinastine which caused QTc prolongation were 85, 35 and over than 3600 ng/ml, respectively. These drugs did not alter the PQ and QRS intervals and did not cause arrhythmia or atrioventricular block. Our results are consistent with the clinical observation that prolongation of QTc is caused by terfenadine and astemizole but not by epinastine. Thus, measurement of QTc in cynomolgus monkey appears to be a useful approach for evaluating the potential cardiotoxicity of histamine H1 receptor antagonists. PMID- 10478630 TI - Marked dissociation between intracellular Ca2+ level and contraction on exposure of rat aorta to lysophosphatidylcholine. AB - We investigated the relationship between tension development and the cytosolic free Ca2+ level ([Ca2+]i) on exposure of the endothelium-denuded isolated rat aorta to palmitoyl-L-alpha-lysophosphatidylcholine. Lysophosphatidylcholine concentration-dependently induced a gradual increase in [Ca2+]i. Application of 10(-4) M lysophosphatidylcholine induced a large and sustained tonic increase in [Ca2+]i (the peak [Ca2+]i was 125.2 +/- 11.5% of the 80 mM K+-induced response) but only a small contraction (4.0 +/- 1.4% of the 80 mM K+ induced contraction). The sustained increase in [Ca2+]i was attenuated when extracellular Ca2+ was removed but it was unaffected by verapamil or 1-(5-isoquinolinesulphonyl)-2 methylpiperazine dihydrochloride (H-7). Digitonin also produced a gradual increase in [Ca2+]i but with a pronounced contraction. Triton X-100 (0.1%) produced a marked elevation in [Ca2+]i with no detectable contraction. Triton X 100, however, caused a rapid leakage of fura PE-3. Treatment with 10(-4) M lysophosphatidylcholine for 1 or 2 h did not affect the contractile response induced by 80 mM K+ and this treatment did not release lactate dehydrogenase from the rat aorta. Treatment with lysophosphatidylcholine did not affect either the cyclic AMP level or the cyclic GMP level in endothelium-denuded aortic tissues. These results show that in the rat aorta lysophosphatidylcholine produces a large increase in [Ca2+]i (possibly in a non-contractile compartment) which does not induce contraction. Thus, the increase in [Ca2+]i induced by lysophosphatidylcholine (i) requires external Ca2+ but is not due to an increased Ca2+ influx through voltage-dependent L-type Ca2+ channels, (ii) is not primarily due to protein kinase C activation and (iii) is probably not due to a detergent action (like those of digitonin and triton X-100). The relative lack of a contractile response to lysophosphatidylcholine is not due to formation of cyclic AMP or cyclic GMP. PMID- 10478631 TI - Correlation between changes in morphology, electrical properties, and angiotensin converting enzyme activity in the failing heart. AB - Evidence is available that morphologic and electrophysiologic abnormalities are present in the failing heart. In the present work, the progressive changes in electrical properties and morphology of the failing heart of Syrian cardiomyopathic hamsters (TO2) were investigated at different stages of the pathological process, and the possible role of the renin-angiotensin system was studied. Cardiomyopathic hamsters 2 and 11 months of age were used. Age-matched normal hamsters (F1B) were utilized as controls. Measurements of membrane potential, conduction velocity and refractoriness were made with conventional intracellular electrodes connected to a high impedance DC amplifier. Serum and cardiac angiotensin-converting enzyme (ACE) activities were measured in controls and cardiomyopathic animals. The results indicated that interstitial fibrosis and calcification were present in the heart of 2-month old Syrian cardiomyopathic hamsters. Measurements of the resting potential performed in the isolated right ventricle of 2-month old Syrian cardiomyopathic hamsters indicated an average value of -66.7 +/- 0.96 mV (n = 25); in the controls of the same age was -78.5 +/ 1 mV (n = 25, P < 0.05); and in 11-month old cardiomyopathic hamsters was -67.8 +/- 0.83 mV (n = 10). The duration of the action potential measured at 50 and 90% of repolarization in 2-month old hamsters was well above the controls. The conduction velocity measured in the isolated right ventricle of 2-month old Syrian cardiomyopathic hamsters (44.2 +/- 1.6 cm/s, n = 12) was not different from the control (43.7 +/- 1.1 cm/s, n = 7, P > 0.05) but was significantly larger than that recorded from the ventricle of 11-month old animals (37.8 +/- 2.9 cm/s, n = 11, P < 0.05). ACE activity was 0.26 +/- 0.01 nmol/mg x min in the heart of controls at 2 months of age and did not change with age. Although in the 2-month old cardiomyopathic hamsters the enzyme activity (0.28 +/- 0.04 nmol/mg x min) was not different from the controls (P > 0.05), in myopathic animals at 11 months of age, the enzyme activity (0.56 +/- 0.027 nmol/mg x min) was greater than controls (P < 0.05). The ACE activity in plasma followed the same pattern. The conclusion from these experiments is, that some parameters like resting potential, action potential duration, and morphological abnormalities appeared quite early in the failing process. The decline in conduction velocity, however, appeared later on, concurrently with the activation of plasma and cardiac renin angiotensin systems. PMID- 10478632 TI - Interactions between AT1 and AT2 receptors in uterine arteries from pregnant ewes. AB - This study was performed to investigate the roles of angiotensin receptors (AT1 and AT2) in the contractility of uterine arteries during normal pregnancy and after angiotensin II levels have been elevated. Pregnant ewes were given intravenous infusions of saline for 24 h (control) or angiotensin II (30 ng kg( 1) min(-1)) for 2 or 24 h. The contractile responses of uterine arterial rings to angiotensin II (4 microM) and antagonists were then examined in vitro. Most uterine arteries were relatively insensitive to the vasoconstrictor effects of angiotensin II. In rings from control ewes an angiotensin AT2 antagonist enhanced (P < 0.05) the contractile responses to angiotensin II, suggesting that angiotensin AT2 receptors inhibited the angiotensin AT1 receptor mediated contractions. Uterine arterial rings from ewes given intravenous infusions of angiotensin II displayed greater (P < 0.05) contractile responses to angiotensin II in vitro compared to rings from control ewes. This was in part due to down regulation of angiotensin AT2 receptors. Surprisingly, while performing these experiments a small number of ewes had uterine arteries which were "hyperreactive" to angiotensin II (contractile responses 6-fold greater). These ewes also had abnormal renin angiotensin systems and had some features which are characteristic of those seen in preeclampsia. The "hyperreactivity" of these arteries could only in part be explained by down regulation of angiotensin AT2 receptors. It is concluded that in normal pregnancy angiotensin AT2 receptors play a role in maintaining an adequate uterine blood flow for the fetus. When angiotensin II levels are elevated for a prolonged period this protective effect is lost partly because angiotensin AT1 receptors are down regulated. PMID- 10478633 TI - Differential regulation of cyclooxygenase isoenzymes by cAMP-elevating agents. AB - Bovine aortic endothelial cells produce prostacyclin as their major arachidonic acid metabolite. cAMP, in turn, is the second messenger for prostacyclin. In the present study, we investigated the effects of cAMP-elevating agents on prostacyclin production by bovine aortic endothelial cells. Treatment of resting bovine aortic endothelial cells with cAMP-elevating agents inhibited prostacyclin production and cyclooxygenase activity, without affecting arachidonic acid release. No change was detected in cyclooxygenase-1 protein expression. The specific inhibitor of protein kinase A, Rp-cAMPS (adenosine 3',5'-cyclic monophosphorothioate, Rp-isomer, triethylammonium salt), and the phosphatase inhibitor, okadaic acid, both suppressed cAMP-induced inhibition, suggesting that this inhibition is mediated by a phosphorylation-dephosphorylation cascade, which is possibly protein kinase A-dependent. In lipopolysaccharide-treated cyclooxygenase-2 expressing bovine aortic endothelial cells, where cyclooxygenase 1 activity was selectively inhibited, dibutyryl cAMP failed to inhibit cyclooxygenase-2 activity. Cyclooxygenase-2 protein was induced upon treatment with dibutyryl cAMP and further induction of cyclooxygenase-2 protein was effected by IBMX (3-isobutyl-1-methyl-xanthine) and dibutyryl cAMP in bacterial lipopolysaccharide-stimulated cells. These results suggest that increased cellular cAMP selectively inhibits cyclooxygenase-1 activity without altering cyclooxygenase-1 protein expression, and at the same time, up-regulates cyclooxygenase-2 protein. This complex regulation of cyclooxygenase activity and protein expression by cAMP may represent a prostacyclin-induced autoregulatory mechanism in bovine aortic endothelial cells. PMID- 10478634 TI - Extraction of active clonidine-displacing substance from bovine lung and comparison with clonidine-displacing substance extracted from other tissues. AB - Crude methanolic clonidine-displacing substance (CDS) extracted from bovine lung competed for radioligand binding from alpha2-adrenoceptors and I2-sites present in rat brain membranes, and from I1-sites present in rat brain and kidney membranes. There was no difference in the competition of [3H]clonidine binding to alpha2-adrenoceptors present in either rat or rabbit brain membranes by the crude CDS extract and therefore either tissue could be used to estimate the number of units of CDS present in extracts. Further purification by reverse phase high performance liquid chromatography (RP-HPLC), with UV detection, of extracts obtained from bovine lung, brain and rat brain exhibited similar three-peak profiles, previously reported. Corresponding fractions competed for radioligand binding to alpha2-adrenoceptors present in rat brain membranes, eluting between 19 and 23 min, which corresponded with the middle peak of the three-peaks. Therefore, we propose the CDS-like material eluting from all these tissues to be similar. Interestingly, CDS extracted from bovine adrenal glands under the same conditions showed a similar three-peak profile, but did not repeat the displacement of binding just at 19-23 min, but at every time point after 4 min. This suggests this tissue could represent a source of CDS in this species. PMID- 10478635 TI - Effects of isoflurane and hexafluorodiethyl ether on human recombinant GABA(A) receptors expressed in Sf9 cells. AB - Effects of volatile anesthetics and a volatile convulsant on human recombinant gamma-aminobutyric acid (GABA) type A receptor responses were studied using the whole cell configuration of the patch clamp technique. Sf9 cells were transfected with bacuroviruses carrying cDNAs of alpha1beta2, alpha1beta2gamma2s, alpha3beta2 and alpha3beta2gamma2s subunit combinations of the human GABA(A) receptor. Clinical concentrations of isoflurane (a volatile anesthetic) enhanced the GABA induced current of the alpha1beta2gamma2s and alpha3beta2gamma2s GABA(A) subunit combinations. On the other hand, isoflurane suppressed the current of the alpha1beta2 and alpha3beta2 subunit combinations, indicating that the anesthetic effects depended upon the presence of gamma2s subunit. A high concentration (2 mM) of isoflurane generated a surge current following the washout of GABA and the anesthetic. Hexafluorodiethyl ether (a volatile convulsant) decreased the GABA response of the both alpha3beta2gamma2s and alpha3beta2 constructs without generating a surge current. The results suggest that volatile agents affect the receptor-ionophore complex via direct interaction with proteins but not through a perturbation of the membrane lipid environment. A hypothetical sequential model for the anesthetic action is presented. PMID- 10478636 TI - Sphingomyelin inhibits platelet 12-lipoxygenase activity. AB - The effect of sphingomyelin on the formation of 12-hydroxy-5,8,10,14 eicosatetraenoic acid (12-HETE), thromboxane B2 and 12-hydroxy-5,8,10 heptadecatrienoic acid (HHT) in washed rabbit platelets was examined. Sphingomyelin had a powerful inhibitory effect on 12-HETE formation, while it produced only a small increase in thromboxane B2 and HHT formation. The sphingomyelin metabolite ceramide did not affect the formation of 12-HETE, thromboxane B2 and HHT. These results suggest that sphingomyelin is a selective inhibitor of platelet 12-lipoxygenase and may have functional effects in platelets. PMID- 10478637 TI - Excitotoxic injury profiles of low-affinity kainate receptor agonists in cortical neuronal cultures. AB - Neurotoxic profiles of putative agonists for low-affinity kainate subtypes of L glutamate receptors (GluR5-7) were determined in cultured cortical neurones. Rank order of neurotoxic potency (microM): (S)-5-iodowillardiine (9) approximately = (2S,4R,6E)-2-amino-4-carboxy-7-(2-naphthyl)hept-6-enoic acid (LY339434, 11) > (2S,4R)-4-methylglutamate (33) > kainate (100) > (RS)-2-amino-3-(hydroxy-5-tert butylisoxazol-4-yl)propanoic acid (ATPA, 360). Using ionotropic glutamate receptor antagonists, neurotoxicity induced by kainate, ATPA and (S)-5 iodowillardiine appeared to involve a GluR5-7 component, unlike LY339434 and (2S,4R)-4-methylglutamate. These putative GluR5-7 agonists exhibited complex excitotoxic profiles highlighting the importance of studying native glutamate receptors. PMID- 10478638 TI - Pharmacokinetics and stability of the ch14.18-interleukin-2 fusion protein in mice. AB - The fusion protein formed from ch14.18 and interleukin-2 (ch14.18-IL-2), shown to exhibit antitumor efficacy in mouse models, consists of IL-2 genetically linked to each heavy chain of the ch14.18 chimeric anti-GD2 monoclonal antibody. The purpose of this study was to determine the pharmacokinetics of ch14.18-IL-2 in mice and assess its stability in murine serum. Following i.v. injection, the fusion protein was found to have a terminal half-life of 4.1 h. Detection of IL-2 following injection of the ch14.18-IL-2 fusion protein showed a similar half life, indicating that the fusion protein prolongs the circulatory half-life of IL 2. Detection of human IgG1 following injection of ch14.18-IL-2 showed a terminal half-life of 26.9 h. These data suggested that the native fusion protein is being altered in vivo, resulting in a somewhat rapid loss of detectable IL-2, despite prolonged circulation of its immunoglobulin components. In vitro incubation of the ch14.18-IL-2 fusion protein in pooled mouse serum at 37 degrees C for 48 h resulted in a loss of its IL-2 component, as detected in enzyme-linked immunosorbent assay systems and in proliferation assays. Polyacrylamide gel electrophoresis and Western blot analysis of the fusion protein incubated in mouse serum at 37 degrees C indicated that the ch14.18-IL-2 is cleaved, resulting in a loss of the 67-kDa band (representing the IL-2 linked to the IgG1 heavy chain) and the detection of a band of more than 50 kDa, slightly heavier than the IgG1 heavy chain itself. This suggests that the fusion protein is being cleaved in vitro within the IL-2 portion of the molecule. These studies show that (1) ch14.18-IL-2 prolongs the circulatory half-life of IL-2 (compared to that of soluble IL-2) and (2) the in vivo clearance of the fusion protein occurs more rapidly than the clearance of the ch14.18 antibody itself, possibly reflecting in vivo cleavage within the IL-2 portion of the molecule, resulting in loss of IL-2 activity. PMID- 10478640 TI - Establishment of gp100 and MART-1/Melan-A-specific cytotoxic T lymphocyte clones using in vitro immunization against preselected highly immunogenic melanoma cell clones. AB - The induction of an in vitro T cell response against tumour-associated antigens with subsequent expansion of the individual cytotoxic T lymphocyte (CTL) clones still is not routine and the only tumour-associated antigen that has been found to easily induce the establishment of CTL clones is the MART-1/Melan-A antigen. In this paper, we describe a new approach for in vitro immunization based on the use of preselected melanoma cell clones. The human melanoma cell subline FM3.P was cloned and the immunological properties of individual clones were compared. Melanoma cell clone FM3.29, having a high level of expression of melanoma differentiation antigens, as well as high levels of the HLA class I and class II antigens and adhesion molecules, was used for the establishment of a CTL line that was subsequently cloned. For optimization of the conditions of growth of established CTL clones, a particular melanoma subline FM3.D/40 was selected for supporting the proliferation of CTL clones. The majority of the established CTL clones recognized the melanoma-associated differentiation antigens gp100 and MART 1/Melan-A. Epitope analysis indicated that two different epitopes derived from gp100 (154-162 and 280-288) and a single epitope from MART-1/Melan-A (27 35) were recognized by these CTL clones. The gp100-specific CTL clones were found to be significantly more sensitive to the culture conditions than the MART-1/Melan-A specific CTL clones. In addition, the presence of excess peptide in the culture medium induced autokilling of the gp100-specific, but not the MART-1/Melan-A specific CTL clones. Taken together, these results demonstrate that, by careful preselection of melanoma cell lines and clones both for the induction of CTL line from patients' peripheral blood lymphocytes and subsequent cloning, it is possible to obtain a large number of stable CTL clones even against such an inherently "difficult" differentiation antigen as gp100. PMID- 10478639 TI - Induction of antitumor immunity after cure of pulmonary metastases, using staphylococcal enterotoxin B and bispecific antibody. AB - The combination of staphylococcal enterotoxin B (SEB) and anti-p97 x anti-CD3 bispecific antibody (bsAb) cures 60%-80% of mice with established pulmonary metastases of the syngeneic p97+ murine melanoma, CL62. We investigated the ability of cured mice to generate protective antitumor immunity. In tumor rechallenge experiments, CL62-cured mice developed protective immunity against rechallenge with CL62. The majority of mice also rejected the p97-negative parental cell line, K1735, indicating an immune response to tumor antigens common to both cell lines that were not bsAb-targeted. A significant humoral response developed against p97 antigen, but not against other antigens common to both CL62 and K1735. That the majority of cured mice nevertheless rejected K1735 suggests that tumor immunity is not antibody-dependent. Evidence of cellular immunity was obtained from the results of delayed-type hypersensitivity, proliferation and cytotoxicity assays, which revealed the presence of tumor-specific memory in bsAb treated, CL62-cured mice. CD8+ T cells from cured, but not control mice were able to lyse tumor; however, memory CD4 cells had no cytolytic function. In vivo, however, both CD4 and CD8 T cells were required for effective protective immunity. These studies demonstrate that treatment with SEB and bsAb not only confers passive immune effects of tumor eradication, but also actively promotes the generation of a host antitumor immune response. PMID- 10478641 TI - Generation of anti-allogeneic cytotoxic T lymphocytes depends upon a signal, not delivered by tumor cells, that can be provided in trans in the presence of exogenous interleukin-2. AB - We have reported that, while immune responses were generated initially against CMS5 tumor cells, they were lost with time. In the present work, we asked whether the tumor cells contributed to this by delivering an inhibitory signal or whether a positive signal, delivered in the form of tumor-derived interleukin-2 (IL-2) or the presence of professional antigen-presenting cells (APC) would be sufficient to enable a sustained response. We observed that the presence of tumor cells did not inhibit the generation of anti-(class I) cytotoxic T lymphocytes (CTL), suggesting that the tumor cells were not directly suppressive. In addition, the lack of a response could not be attributed to insufficient levels of IL-2 alone, since even tumor cells that secreted IL-2 failed to stimulate anti-(class I) CTL. Finally, we observed that professional APC were necessary to deliver an essential signal, but only in the presence of IL-2-secreting tumor cells. Thus, CMS5 cells failed to provide essential signals necessary for CTL generation, but were not directly suppressive. PMID- 10478642 TI - On down-regulation of the immune response to metastatic malignant melanoma. AB - Treatment of metastatic malignant melanoma with interferon alpha (IFNalpha) results in objective remission in approximately 15% of patients. In a previous investigation, we found that about 50% of the patients achieved at least minor or short-lived remissions. In some tumours extensive areas of regressive tumour change occurred. However, even in these areas remnants of tumour cells were generally found. The short duration of the immune response in some patients and the incomplete eradication of the tumour can be due either to selection of non immunogenic tumour cells or to down-regulation of the immune reactivity to the tumour. In the present paper, the expression of the zeta chain of the T cell receptor in CD3+ lymphocytes and the expression of CD28 in CD3+, CD4+ and CD8+ lymphocytes was studied in resectable melanoma metastases from 20 treated (IFNalpha or IFNalpha in combination with cisplatinum and dacarbazine) and 16 untreated patients. A double-staining technique was used, and the occurrence and distribution of lymphocytes showing down-regulation of the zeta chain or CD28 were separately registered in different areas of the metastases: close to the tumour cells in areas of unaffected tumour growth, in areas with regressive tumour changes, in areas with marked fibrosis and in stromal areas with densely packed lymphocytes. CD3+ zeta lymphocytes were found in all metastases, but their number and distribution varied considerably. Down-regulation of the zeta chain was most often found in areas of regressive changes. In contrast, T lymphocytes infiltrating close to the tumour cells had a stronger expression of the zeta chain (P = 0.016). Down-regulation was also found in stromal areas of densely packed lymphocytes and in areas of fibrosis. The pattern of down-regulation of CD28 in various subsets of lymphocytes was similar to that of zeta chain. The same pattern of down-regulation of CD28 and the zeta chain was found in both untreated and treated patients, indicating that the down-regulation is not due to treatment but to the release of immunosuppressor factors from areas with high tumour cell density or extensive destruction of tumour cells. These results concur well with the view that IFNalpha treatment can result in immune-mediated tumour cell destruction early in the treatment period and that this immune response to the tumour can be followed by immunosuppression within a few weeks. PMID- 10478643 TI - Restoration of expression of signal-transduction molecules in lymphocytes from patients with metastatic renal cell cancer after combination immunotherapy. AB - A decrease in lymphocyte signal-transduction molecules, described in cancer patients and patients with chronic infectious diseases, has been proposed as a possible mechanism leading to an impaired immune response in cancer patients. Here we report the effects of combination immunotherapy on the levels of T cell receptor zeta chain and p56lck tyrosine kinase in a retrospective study of cryopreserved lymphocytes from 26 metastatic renal cell carcinoma patients treated with high-dose interleukin-2 (IL-2), interferon alpha (IFNalpha) and ex vivo IL-2-activated lymphocytes. Of the 26 patients, 12 were responders (5 complete and 7 partial) and 14 were non-responders (6 stable and 8 with progressive disease). Prior to treatment, 21 of 26 patients (81%) and 13 of 21 patients (62%) respectively expressed zeta chain and p56lck at less than 50% of the levels observed in healthy controls. During therapy, this low zeta chain and p56lck expression increased to at least 50% of normal in 13 of the 21 patients (62%) and in 6 of the 13 patients (46%) respectively; in the remaining patients expression levels remained at 50% of normal or more, or declined. Although, in this limited study, pretreatment levels of and p56lck did not show significant correlation with antitumor response, 4 of 5 patients that achieved a complete response (80%) corrected both zeta chain and p56lck levels to at least 50% of normal, while restoration of both signal-transduction molecules to such levels was only observed in 3 of 7 partial responders (43%), 1 of 5 patients with stable disease (20%) and 2 of 7 patients with progressive disease (29%). Thus, these results suggest that analysis of changes in signal-transduction molecules may a be useful tool for immunological monitoring of patients throughout immunotherapy, and could provide important information for designing new clinical trials that restore impaired signal transduction while activating T cell responses. PMID- 10478644 TI - Immunoregulation in pancreatic cancer patients. AB - Metastatic pancreatic cancer is one of the most aggressive cancer known in man yet specific antitumor immunity has been demonstrated in lymph nodes draining the sites of pancreatic tumors. Despite this immunity, pancreatic cancer patients suffer a quick demise. To further define tumor immunity in patients with metastatic pancreatic cancer, we sought to characterize helper T cell subsets, serum cytokines, cellular cytotoxicity that is both T-cell and non-T cell mediated, as well as known tumor-derived immunosuppressive products that may be present in their peripheral blood. Significantly heightened levels of interleukin 2 (IL-2), a Th1 cytokine, were found in patients before treatment with chemotherapy while serum IL-10, a Th2 cytokine, were at significantly lower levels than observed in normal donors tested between their fifth and seventh decades of life. IL-10 levels increased progressively with age as a serum-bound protein in normal, healthy donors tested between the ages of 24 through 61. An age associated progression of increased IL-10 levels was not observed in pancreatic cancer patients. Few patients had detectable serum levels of soluble fas ligand but approximately half had elevated levels of a tumor marker, detected with the CA-15.3 assay, known as soluble MUCIN 1 (MUC1). Cell mediated cytotoxicity including T-cell mediated killing of pancreatic tumor cell lines was detected in many patients. These data suggest that pancreatic cancer patients have activated type 1 helper T cells that can support development of cell mediated immunity, and that their sera contain lowered levels of the "anti inflammatory" type 2 cytokine, IL-10. PMID- 10478645 TI - Altered in vitro and in vivo flumazenil binding in human epileptogenic neocortex. AB - In vitro and in vivo parameters of flumazenil (FMZ) binding were measured in spiking and nonspiking neocortex identified by intraoperative electrocorticography in epileptic patients who underwent cortical resection for seizure control. In vitro measures of receptor affinity (K(D)), number (Bmax) and laminar distribution for [3H]-FMZ binding in the epileptic focus (n = 38) were compared to nonspiking cortex from a subgroup of the patients (n = 12) and to tissue obtained from trauma patients (n = 5). The in vitro binding parameters were compared to in vivo [11C]-FMZ binding measured with positron emission tomography (PET) (n = 19). The Bmax was higher in the 38 spiking tissues as compared to the 12 nonspiking tissues (P = .012). Paired comparison of spiking versus nonspiking binding in the 12 patients from whom nonspiking tissue was available showed increases in both K(D) (P = .037) and Bmax (P = .0047) in spiking cortex. A positive correlation was found between K(D) and Bmax values for 38 patients (r = 0.55, P < .0001), the magnitude of the K(D) increase being twice that of the Bmax increase. In addition, there was a significant correlation between the asymmetry indices of the in vivo FMZ binding on PET and in vitro K(D) of spiking cortex (n = 19, r = 0.52, P = .02). The laminar distribution of [3H] FMZ showed increased FMZ binding in cortical layers V-VI in spiking cortex compared to nonspiking and control cortex. The increased receptor number in spiking cortical layers V-VI may be a compensatory mechanism to decreased GABAergic input. The increased Bmax in spiking cortex was accompanied by a larger decrease in the affinity of FMZ for the receptor suggesting that decreased FMZ binding in the epileptic focus measured with PET is due to a decrease in the affinity of the tracer for the receptor. PMID- 10478646 TI - Blood-brain barrier transport and protein binding of flumazenil and iomazenil in the rat: implications for neuroreceptor studies. AB - The calculated fraction of receptor ligands available for blood-brain barrier passage in vivo (f(avail)) may differ from in vitro (f(eq)) measurements. This study evaluates the protein-ligand interaction for iomazenil and flumazenil in rats by comparing f(eq) and f(avail). Repeated measurements of blood-brain barrier permeability for two benzodiazepine antagonists were performed in 44 rats by the double-indicator technique. Cerebral blood flow was measured by intracarotid Xe-injection. The apparent permeability-surface product (PSapp) was measured while CBF or bolus composition was changed. Comparison of PSapp obtained in the absence and presence of 5% albumin in the injectate yielded f(avail), whereas f(eq) was measured by equilibrium dialysis. Iomazenil and flumazenil f(avail) was 62% and 82%, respectively, whereas f(eq) was significantly lower, 42% and 61%. The PSapp for iomazenil and flumazenil increased significantly by 89% and 161% after relative CBF increases of 259% and 201%, respectively. The results demonstrate that application of f(eq) in neuroreceptor studies underestimates the plasma input function to the brain. Model simulations render possible that the differences between f(avail) and f(eq) as well as the effect of CBF on PSapp can be caused by capillary heterogeneity. PMID- 10478647 TI - Quantification of delta-opioid receptors in human brain with N1'-([11C]methyl) naltrindole and positron emission tomography. AB - The regional binding of N1'-([11C]methyl)naltrindole (MeNTI), a selective delta opioid antagonist, was studied in healthy human subjects with positron emission tomography (PET). After the bolus intravenous administration of high specific activity [11C]MeNTI, PET was performed over 90 minutes. Arterial plasma samples were obtained during the scanning period and assayed for the presence of radiolabeled metabolites. The data were analyzed with various kinetic (two- and three-compartment models, Patlak graphical analysis) and nonkinetic (apparent volume of distribution and activity at a late scanning time) approaches. This tracer showed irreversible binding characteristics during the scanning period used. The results of the analyses also were compared with the density and distribution of delta-opioid receptors in the human brain in vitro. Additionally, computer simulations were performed to assess the effects of changes in receptor binding and tracer transport changes on the perceived binding parameters obtained with the models. A constrained three-compartment kinetic model was demonstrated to be superior to other quantification models for the description of MeNTI kinetics and quantification of delta receptor binding in the human brain with 11C labeled MeNTI. PMID- 10478649 TI - Butanol is superior to water for performing positron emission tomography activation studies. AB - [15(O)]Butanol has been shown to be superior to [15(O)]water for measuring cerebral blood flow with positron emission tomography. This work demonstrates that it is also superior for performing activation studies. Data were collected under three conditions: a visual confrontation animal-naming task, nonsense figure size discrimination, and a nonvisual darkroom control task. Time-activity curves (TAC) were obtained for regions known to be activated by the confrontation naming task to compare absolute uptake and the different kinetics of the two tracers. Also, t statistic maps were calculated from the data of 10 subjects for both tracers and compared for magnitude of change and size of activated regions. Peak uptake in the whole-brain TAC were similar for the two tracers. For all regions and conditions, the washout rate of [15(O)]butanol was 41% greater than that of [15(O)]water. At a threshold of 0, the [15(O)]water and [15(O)]butanol percent difference (nonnormalized) and t statistic (global normalization) images are nearly identical, indicating that the same property is being measured with both tracers. The [15(O)]butanol parametric images displayed at a threshold of /t/ = 5 look similar to the [15(O)]water parametric maps displayed at a threshold of /t/ = 4, which is consistent with the observation that t statistic values in [15(O)]butanol images are generally greater. The t statistic values were equal when the [15(O)]butanol parametric map was created from any subset of 6 subjects and the [15(O)]water parametric map was created from all 10 subjects. Fewer subjects need to be studied with [15(O)]butanol to reach the same statistical power as an [15(O)]water-based study. PMID- 10478650 TI - Cerebral vasodilatation causing acute intracranial hypertension: a method for noninvasive assessment. AB - Deep spontaneous vasodilatatory events are frequently recorded in various cerebral diseases, causing dramatic increases (A-waves) in intracranial pressure (ICP) and subsequently provoking ischemic brain insults. The relationship between fluctuations in CBF, ICP, and arterial blood pressure (ABP) is influenced by properties of cerebrovascular control mechanisms and the cerebrospinal pressure volume compensation. The goal of this study was to construct a mathematical model of this relationship and to assess its ability to predict the occurrence and time course of A-waves. A group of 17 severely head-injured patients were included in the study. In our model ICP was derived from the ABP waveform using a linear signal transformation. The transformation was modified during the simulation by a relationship between ABP and flow velocity, i.e., by the characterization of the cerebrovascular bed. In this way the ICP could be calculated from the ABP waveform. This model was verified by comparison of simulated and directly measured ICP during A-waves recorded in seven of the patients. In all simulations, plateau elevations of ICP were well replicated. The mean absolute error between real and simulated ICP was 8.3 +/- 5.4 mm Hg at the baseline and 7.9 +/- 4.3 mm Hg at the top of plateau waves. The correlation coefficient between real and simulated increase in ICP was R = 0.98; P < .001. Similarly, correlation between real and simulated increase in pulse amplitude of ICP was highly significant (R = 0.94; P < .001). The mathematical model of the relationship between ABP, flow velocity, and ICP is of potential clinical use for the noninvasive detection of A-waves in patients in whom invasive ICP assessment is not conducted. PMID- 10478648 TI - Kinetic analysis of [11C]McN5652: a serotonin transporter radioligand. AB - The impulse response function of a radioligand is the most fundamental way to describe its pharmacokinetics and to assess its tissue uptake and retention pattern. This study investigates the impulse response function of [11C](+)McN5652, a radioligand used for positron emission tomography (PET) imaging of the serotonin transporter (SERT) in the brain. Dynamic PET studies were performed in eight healthy volunteers injected with [11C](+)McN5652 and subsequently with its pharmacologically inactive enantiomer [11C](-)McN5652. The impulse response function was calculated by deconvolution analysis of regional time-activity curves, and its peak value (f(max)), its retention value at 75 minutes (fT), and its normalized retention (f(rel) = fT/f(max)) were obtained. Alternatively, compartmental models were applied to calculate the apparent total distribution volume (DV(T)) and its specific binding component (DV(S)). Both the noncompartmental (fT,f(rel)) and the compartmental parameters (DV) were investigated with and without correction for nonspecific binding by simple subtraction of the corresponding value obtained with [11C](-)McN5652. The impulse response function obtained by deconvolution analysis demonstrated high tracer extraction followed by a slow decline in the form of a monoexponential function. Statistical analysis revealed that the best compartmental model in terms of analysis of variance F and condition number of the parameter variance-covariance matrix was the one that was based on a single tissue compartment with parameters k1 and k2 and that also included the parameter of regional cerebral blood volume (BV). The parameter f(rel) demonstrated low between-subject variance (coefficient of variation [CV] = 19%), a midbrain to cerebellum ratio of 1.85, and high correlation with the known density of SERT (r = 0.787 where r is the coefficient of linear correlation between the parameter and the known density of SERT). After correction for nonspecific binding, f(rel) demonstrated further improvement in correlation (r = 0.814) and midbrain to cerebellum ratio (3.09). The variance of the distribution volumes was acceptable when the logarithmic transform lnDV was used instead of DV (17% for the three-parameter model), but correlation of this compartmental parameter was slightly less (r = 0.652 for the three-parameter model) than the correlation of the noncompartmental f(rel) with the known density of SERT, and the midbrain to cerebellum ratio was only 1.5 (uncorrected) and 1.8 (corrected). At the expense of increasing variance, the correlation was increased after correction for nonspecific binding using the inactive enantiomer (r = 0.694; CV = 22%). These results indicate that the kinetics of [11C](+)McN5652 can best be described by a one-tissue compartment model with three parameters (k1, k2, and BV), and that both the noncompartmental parameter f(rel) and the compartmental distribution volumes have the potential for quantitative estimation of the density of SERT. Further validation of the radioligand in experimental and clinical situations is warranted. PMID- 10478651 TI - Early training may exacerbate brain damage after focal brain ischemia in the rat. AB - Early overuse of a lesioned forelimb, induced by immediate immobilization of the intact forelimb after a cortical lesion, has been reported to increase tissue damage and delay functional recovery. To investigate if early training without immobilization of the intact forelimb could increase tissue loss and reduce recovery, the middle cerebral artery was ligated distal to the striatal branches in 25 male spontaneously hypertensive rats. Control rats were housed in standard cages, training rats were transferred to larger cages allowing various activities and received additional special training 1 hour a day starting either 24 hours or 7 days after the ligation. The rats were tested on a rotating pole, in a leg placement test, and in a water maze and they were killed 6 weeks after the ligation. Delayed training resulted in the best overall performance; however, both training groups performed better than standard rats on the rotating pole. The cortical infarct volume was larger in the early training group than in the other two groups (P < .005), possibly related to increased glutamate release and peri-infarct cortical hyperexcitability. PMID- 10478652 TI - Increased cytokine release from peripheral blood cells after acute stroke. AB - Cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha can play pathogenetic or protective roles in stroke. They are increased in the brain after experimental ischemia and in the CSF of patients with stroke. However, their presence in the periphery is still controversial. To determine the source and time-course of cytokines in blood of stroke patients, IL-6 and TNF-alpha release from blood cells and serum levels were determined in 40 patients on days 1 through 2, 4, 10, 30, and 90 after stroke. Twenty healthy age-matched volunteers were used as controls. IL-6 and TNF-alpha release from stimulated blood cells was increased in stroke patients, compared to controls. A peak response (+224%) was observed at day 4 for IL-6, while TNF-alpha release was largely and significantly increased (about three-fold compared to controls) from day 1 to 2 until day 90 after stroke. The increase in IL-6 release was significantly higher in ischemic, compared to hemorrhagic strokes, at days 1 and 4. Circulating IL-6 was increased at each time point. The ischemic processes in the CNS induces a long-lasting activation of IL-6 and TNF-alpha production in peripheral blood cells, which are a major source of serum cytokines after stroke. PMID- 10478653 TI - Insulin induces dephosphorylation of eukaryotic initiation factor 2alpha and restores protein synthesis in vulnerable hippocampal neurons after transient brain ischemia. AB - Brain reperfusion causes prompt, severe, and prolonged protein synthesis suppression and increased phosphorylation of eukaryotic initiation factor 2alpha [eIF2alpha(P)] in hippocampal CA1 and hilar neurons. The authors hypothesized that eIF2alpha(P) dephosphorylation would lead to recovery of protein synthesis. Here the effects of insulin, which activates phosphatases, were examined by immunostaining for eIF2alpha(P) and autoradiography of in vivo 35S amino acid incorporation. Rats resuscitated from a 10-minute cardiac arrest were given 0, 2, 10 or 20 U/kg of intravenous insulin, underwent reperfusion for 90 minutes, and were perfusion fixed. Thirty minutes before perfusion fixation, control and resuscitated animals received 500 microCi/kg of 35S methionine/cysteine. Alternate 30-microm brain sections were autoradiographed or immunostained for eIF2alpha(P). Controls had abundant protein synthesis and no eIF2alpha(P) in hippocampal neurons. Untreated reperfused neurons in the CA1, hilus, and dentate gyrus had intense staining for eIF2alpha(P) and reduced protein synthesis; there was little improvement with treatment with 2 or 10 U/kg of insulin. However, with 20 U/kg of insulin, these neurons recovered protein synthesis and were free of eIF2alpha(P). These results show that the suppression of protein synthesis in the reperfused brain is reversible; they support a causal association between eIF2alpha(P) and inhibition of protein synthesis, and suggest a mechanism for the neuroprotective effects of insulin. PMID- 10478654 TI - Early appearance of activated matrix metalloproteinase-9 after focal cerebral ischemia in mice: a possible role in blood-brain barrier dysfunction. AB - During cerebral ischemia blood-brain barrier (BBB) disruption is a critical event leading to vasogenic edema and secondary brain injury. Gelatinases A and B are matrix metalloproteinases (MMP) able to open the BBB. The current study analyzes by zymography the early gelatinases expression and activation during permanent ischemia in mice (n = 15). ProMMP-9 expression was significantly (P < 0.001) increased in ischemic regions compared with corresponding contralateral regions after 2 hours of ischemia (mean 694.7 arbitrary units [AU], SD +/- 238.4 versus mean 107.6 AU, SD +/- 15.6) and remained elevated until 24 hours (mean 745.7 AU, SD +/- 157.4). Moreover, activated MMP-9 was observed 4 hours after the initiation of ischemia. At the same time as the appearance of activated MMP-9, we detected by the Evan's blue extravasation method a clear increase of BBB permeability. Tissue inhibitor of metalloproteinase-1 was not modified during permanent ischemia at any time. The ProMMP-2 was significantly (P < 0.05) increased only after 24 hours of permanent ischemia (mean 213.2 AU, SD +/- 60.6 versus mean 94.6 AU, SD +/- 13.3), and no activated form was observed. The appearance of activated MMP-9 after 4 hours of ischemia in correlation with BBB permeability alterations suggests that MMP-9 may play an active role in early vasogenic edema development after stroke. PMID- 10478655 TI - Adventitial expression of recombinant endothelial nitric oxide synthase gene reverses vasoconstrictor effect of endothelin-1. AB - The present study was designed to determine the effect of recombinant endothelial nitric oxide synthase (eNOS) gene expression on reactivity of canine basilar arteries to endothelin-1 (ET-1). Experiments were performed ex vivo. The arteries were exposed (30 minutes at 37 degrees C) to adenoviral vectors encoding eNOS gene (AdCMVeNOS) or beta-galactosidase reporter gene (AdCMVbeta-Gal). Twenty-four hours after transduction, transgene expression was evident mainly in the vascular adventitia. Rings of control (nontransduced), AdCMVbeta-Gal- and AdCMVeNOS transduced arteries with and without endothelium were suspended for isometric tension recording. Levels of guanosine 3',5'-cyclic monophosphate (cGMP) were measured by radioimmunoassay. During contractions to uridine 5'-triphosphate, ET 1 (10(-10) to 3x10(-9) mol/L) caused further increase in tension in control and AdCMVbeta-Gal-transduced arteries. In contrast, ET-1 caused concentration dependent relaxations of AdCMVeNOS-transduced arteries. The relaxations to ET-1 in AdCMVeNOS-transduced arteries were endothelium-independent. They were abolished by N(G)-nitro-L-arginine methyl ester or by chemical treatment of adventitia with paraformaldehyde before gene transfer. ET-1 (10(-9) mol/L) significantly increased intracellular cGMP levels in AdCMVeNOS-transduced arteries without endothelium. In arteries transduced with AdCMVeNOS, higher concentrations (10(-9) to 3x10(-8) mol/L) of ET-2 also caused relaxations, whereas ET-3 and sarafotoxin, a selective ET(B) receptor agonist, did not produce any relaxations. The relaxations to ET-1 in AdCMVeNOS-transduced arteries were strongly reduced by BQ-123 (10(-7) mol/L), an ET(A) receptor antagonist, but were not affected by BQ-788 (3x10(-7) mol/L), an ET(B) receptor antagonist. These results suggest that genetically modified adventitia can produce nitric oxide and cause relaxations in response to ET-1 via activation of ET(A) receptors. Our findings support a novel concept that successful transfer and expression of recombinant eNOS gene can lead to a qualitative change in responsiveness to vasoconstrictor substances. PMID- 10478656 TI - Activated microvessels express vascular endothelial growth factor and integrin alpha(v)beta3 during focal cerebral ischemia. AB - Both vascular endothelial growth factor (VEGF) and integrin alpha(v)beta3 play roles in angiogenesis. In noncerebral vascular systems, VEGF can induce endothelial integrin alpha(v)beta3 expression. However, it is unknown whether VEGF, like integrin alpha(v)beta3, appears in the initial response of microvessels to focal brain ischemia. Their coordinate expression in microvessels of the basal ganglia after middle cerebral artery occlusion (MCAO) in the nonhuman primate model was examined quantitatively. Cells incorporating deoxyuridine triphosphate (dUTP+) by the polymerase I reaction at 1 hour (n = 3), 2 hours (n = 3), and 7 days (n = 4) after MCAO defined the ischemic core (Ic) and peripheral regions. Both VEGF and integrin alpha(v)beta3 were expressed by activated noncapillary (7.5- to 30.0-microm diameter) microvessels in the Ic region at 1 and 2 hours after MCAO. At 7 days after MCAO, the number of VEGF+, integrin alpha(v)beta3+, or proliferating cell nuclear antigen-positive microvessels had decreased within the Ic region. The expressions of VEGF, integrin alpha(v)beta3, and proliferating cell nuclear antigen were highly correlated on the same microvessels using hierarchical log-linear statistical models. Also, VEGF and subunit alpha(v) messenger ribonucleic acids were coexpressed on selected microvessels. Here, noncapillary microvessels are activated specifically early during a focal cerebral ischemic insult and rapidly express VEGF and integrin alpha(v)beta3 together. PMID- 10478657 TI - Separate neuronal and glial Na+,K+-ATPase isoforms regulate glucose utilization in response to membrane depolarization and elevated extracellular potassium. AB - The role of cell type-specific Na+,K+-ATPase isozymes in function-related glucose metabolism was studied using differentiated rat brain cell aggregate cultures. In mixed neuron-glia cultures, glucose utilization, determined by measuring the rate of radiolabeled 2-deoxyglucose accumulation, was markedly stimulated by the voltage-dependent sodium channel agonist veratridine (0.75 micromol/L), as well as by glutamate (100 micromol/L) and the ionotropic glutamate receptor agonist N methyl-D-aspartate (NMDA) (10 micromol/L). Significant stimulation also was elicited by elevated extracellular potassium (12 mmol/L KCl), which was even more pronounced at 30 mmol/L KCl. In neuron-enriched cultures, a similar stimulation of glucose utilization was obtained with veratridine, specific ionotropic glutamate receptor agonists, and 30 mmol/L but not 12 mmol/L KCl. The effects of veratridine, glutamate, and NMDA were blocked by specific antagonists (tetrodotoxin, CNQX, or MK801, respectively). Low concentrations of ouabain (10( 6) mol/L) prevented stimulation by the depolarizing agents but reduced only partially the response to 12 mmol/L KCl. Together with previous data showing cell type-specific expression of Na+,K+-ATPase subunit isoforms in these cultures, the current results support the view that distinct isoforms of Na+,K+-ATPase regulate glucose utilization in neurons in response to membrane depolarization, and in glial cells in response to elevated extracellular potassium. PMID- 10478658 TI - When is a hyperplastic polyp not a hyperplastic polyp? PMID- 10478659 TI - Papillary intralymphatic angioendothelioma (PILA): a report of twelve cases of a distinctive vascular tumor with phenotypic features of lymphatic vessels. AB - Six childhood vascular tumors were designated as "malignant endovascular papillary angioendothelioma" by Dabska in 1969. Since then, a few reports of similar cases were published, often called "Dabska tumors." Twelve similar cases were identified in review of vascular tumors from the authors' institutions. There were five men and seven women, including seven adults. Patient ages ranged from 8 to 59 years (mean, 30 years). The tumors occurred in the dermis or subcutis of the buttocks or thigh (n = 6), thumb or hand (n = 3), abdomen (n = 2), and heel (n = 1). The tumor sizes ranged from 1 to more than 40 cm (mean, 7.0 cm). The unifying feature of all cases was distinctive intravascular growth of well-differentiated endothelial cells presenting as a matchstick columnar configuration, sometimes with a large production of matrix that was positive for collagen type IV. In half the cases, these intravascular proliferations had an associated actin-positive pericytic proliferation. There was minimal cytologic atypia and rare to absent mitotic activity. Two cases had an adjacent lymphangioma, and two additional cases had clusters of lymphatic vessels adjacent to the tumor. All but two of the cases showed varying degrees of stromal or intraluminal lymphocytes. Occasional epithelioid endothelial cells were seen, but no cases had features typical of epithelioid, spindle cell, or retiform hemangioendothelioma. Tumor cells were positive for vimentin, von Willebrand factor, CD31, and focally for CD34 and were negative for keratins, epithelial membrane antigen, S-100 protein, and desmin. Vascular endothelial cell growth factor receptor type 3, a recently introduced marker for lymphatic endothelia, was positive in all eight cases that were studied, supporting a lymphatic phenotype. Follow-up in 8 of the 12 cases showed no evidence of recurrences, metastases, or residual disease during follow-ups ranging from 1 to 17 years (mean, 9 years). Based on the proliferative borderline features and the lymphatic phenotype, we propose to designate these tumors as papillary intralymphatic angioendothelioma. Additional cases with extensive follow-up should be studied to rule out variants with malignant potential. PMID- 10478660 TI - Renal angiomyolipoma: a clinicopathologic, immunohistochemical, and follow-up study of 46 cases. AB - We reviewed 46 cases of renal angiomyolipoma covering the period from 1977 to 1997. Eight cases were associated with tuberous sclerosis and one with lymphangiomyomatosis. Histologically, the lesions were most often classic with the three usual components, i.e., mature adipose tissue, thick-walled blood vessels, and smooth muscle. Seven cases were particularly misleading: three cases were entirely adipose mimicking liposarcoma: two cases had an exclusively smooth muscle component, one mimicking lymphangiomyomatosis and one with epithelioid cells; another case had a monophasic epithelioid pleomorphic component ("REON": renal epithelioid oxyphilic neoplasm) and proved to be fatal; and another case was associated with collecting duct carcinoma. The immunohistochemical profile showed the coexpression of alpha-smooth-muscle actin and HMB45. Our study is the first to show positivity of estrogen and progesteron receptors or both in more than 25% of cases. Of 35 cases with follow-up information, only one patient died of malignant spread of angiomyolipoma. PMID- 10478661 TI - Histopathologic effects of three-dimensional conformal external beam radiation therapy on benign and malignant prostate tissues. AB - We reviewed 137 prostate sextant needle biopsies from 137 patients obtained at a median of 35.7 months after three-dimensional conformal external beam radiation therapy (3DCRT). Thirty-one patients (23%) received 3 months of androgen deprivation therapy (ADT) before 3DCRT. We also retrospectively reviewed and assigned a combined Gleason score to the pre-3DCRT needle biopsies (97 patients) or transurethral resection of the prostate gland (1 patient). High-molecular weight cytokeratin (34betaE12) and prostate-specific antigen (PSA) immunohistochemistry was performed in select cases. After 3DCRT, histopathologic changes in benign prostate gland consisted of glandular atrophy, cytologic atypia, and basal cell prominence. The benign glands showed intensely positive reactions with antibodies to high-molecular-weight cytokeratin (34betaE12) and negative to weakly positive reactions to PSA. Paneth cell-like change was seen in 44 (32%) of the biopsies, mucinous metaplasia in 29 (21%), luminal blue-tinged mucinous secretions in 14 (10%), and squamous metaplasia in 8 (6%). The changes in benign prostate tissues were similar between patients treated with ADT and 3DCRT and those treated with 3DCRT alone. After 3DCRT, we recognized two histologic patterns of prostate cancer: (1) prostate cancer showing radiation therapy (RT)-related changes characterized by PSA-positive/34betaE12-negative poorly formed glands or individual cells with abundant clear to finely granular cytoplasm, and (2) prostate cancer showing no apparent RT effect. High-grade prostatic intraepithelial neoplasia (PIN) was seen in 12 post-3DCRT biopsies (8.8%). The use of neoadjuvant ADT had a significant impact on the results of post-RT biopsy. Of the 31 patients treated with neoadjuvant ADT and 3DCRT, 3 (10%) had post-3DCRT biopsies showing prostate cancer without RT effect compared to 44 of 106 men (41%) treated with 3DCRT alone (p = 0.004). Compared to the Gleason score pre-RT, the Gleason score of cancers showing no RT effect was the same in 25 patients (71%), +/-1 point in 8 patients (23%), and +2 points in 2 patients (6%). The mean combined Gleason score post-RT was slightly, although significantly, higher than that pre-RT (7.29 +/- 0.71 versus 7.00 +/- 0.59, p = 0.01). Serum PSA at the time of post-3DCRT biopsy correlated with biopsy results. Prostate cancer without therapy effect was seen in only one of 43 patients (2%) with a serum PSA level < or = 1 ng/ml compared to 46 of 94 patients (49%) with a PSA level > 1 ng/ml (p = 0.0001). After 3DCRT, benign prostate glands show profound histopathologic changes and may be confused with prostate cancer. The effects of 3DCRT on prostate cancer are variable, with some cases showing profound therapy-related changes and others showing no apparent therapy effect. PMID- 10478662 TI - Dissecting leiomyomas of the uterus other than cotyledonoid dissecting leiomyomas: a report of eight cases. AB - We report eight cases of benign uterine smooth muscle neoplasms with unusual growth patterns and intramural dissection. All the patients in our series were of reproductive age or perimenopausal (range, 36-51 years) and had an enlarged uterus or a pelvic mass, with the exception of one lesion that was found incidentally in a patient treated for uterine prolapse. Three also had abnormal uterine bleeding. On gross examination, the lesions had an unusual appearance and were often lobulated and irregular with indistinct margins. On microscopic examination of all the lesions in this study, a dominant benign smooth muscle tumor was associated with intramural dissection of the myometrium by fascicles of neoplastic smooth muscle. Of the eight cases showing intramural dissection, four were intramural dissecting leiomyomas; three were examples of intravenous leiomyomatosis; and one was a multinodular leiomyoma with hydropic degeneration. We excluded cotyledonoid dissecting leiomyomas from the study. In two of the three cases of intravenous leiomyomatosis, extrauterine extensions in continuity with the intramural components were noted at surgery and on gross examination. Intramural dissection of the myometrium by a benign smooth muscle tumor is one additional possibility to be considered in the differential diagnosis of leiomyosarcoma and low-grade stromal sarcoma. PMID- 10478663 TI - The histopathology of cutaneous lesions of Kikuchi's disease (necrotizing lymphadenitis): a report of five cases. AB - Kikuchi's disease (KD) is an idiopathic, self-limited necrotizing lymphadenitis that can clinically and histologically mimic high-grade lymphoma, including Hodgkin's disease, or can be mistaken for the lymphadenitis of systemic lupus erythematosus (SLE). Involvement of extranodal sites is unusual but well documented, especially in Asia, where KD is more common than in North America or Europe. The successful distinction of KD from malignant lymphoma and SLE is imperative for the appropriate treatment of affected patients. We describe five patients with cutaneous involvement by KD, all of whom presented with fever, lymphadenopathy, and an eruption on the skin of the upper body, which in one case was clinically suspected to be due to SLE and in another, polymorphous light eruption. The patients ranged in age from 10 months to 42 years (median, 33 years) and included three females and two males. All five patients had negative serologic studies for collagen vascular disease. Each patient had a lymph node biopsy showing the typical necrotizing lymphadenitis of KD. Skin biopsies from all five patients shared a specific constellation of histologic features: vacuolar interface change with necrotic keratinocytes, a dense lymphohistiocytic superficial and deep perivascular and interstitial infiltrate, varying amounts of papillary dermal edema, and abundant karyorrhectic debris with a conspicuous absence of neutrophils and a paucity of plasma cells, paralleling the nodal histology in KD. CD68 immunohistochemistry on paraffin-embedded sections showed many histiocytes and plasmacytoid monocytes in all cases, whereas CD3, CD4, and CD8 showed highly variable staining among the cases. There was only rare staining with TIA-1 and CD30. We believe that the papular eruption of KD has recognizable histopathologic features and that a CD68 stain that marks many cells that initially seem to be lymphocytes can be performed to confirm the diagnosis. PMID- 10478664 TI - Use of keratin 35betaE12 as an adjunct in the diagnosis of mammary intraepithelial neoplasia-ductal type--benign and malignant intraductal proliferations. AB - A variety of studies have investigated the role of low molecular weight (LMW) and high molecular weight (HMW) cytokeratin (CK) expression in the normal breast and invasive breast carcinomas. A few studies with small numbers of cases have addressed this issue in intraductal proliferations of the breast. This study investigates the expression of these CKs in a large series of ductal intraepithelial neoplasias of the breast. We examined 150 ductal carcinomas in situ (DCIS), 35 cases of intraductal hyperplasia (IDH), and 15 cases of atypical intraductal hyperplasia (AIDH). Immunohistochemistry was performed using monoclonal antibodies against CK-34betaE12 (HMW CK), CK-8, and CK-19 (LMW CK) on formalin-fixed, paraffin-embedded tissue. The intensity (0, +1, +2, +3) and percentage of positive intraductal cells (0-100%) were multiplied to obtain a score from 0 to 300. The immunoprofiles of IDH, AIDH, and DCIS were categorized into four groups showing negative or low (0-60), moderate (61-100), high (101 200), and very high (201-300) scores. All cases of IDH showed an intensely positive reaction (high to very high scores) for CK-34betaE12. In contrast, 90% of the DCIS showed a negative or only focal and weak reaction (negative or low score) for this antigen. The remaining 10% of DCIS showed a positive immunoreaction for CK-34betaE12 with moderate to high scores. All cases of florid IDH and 96% of cases of DCIS expressed CK-8 intensely with high to very high scores. Although CK-19 was strongly expressed in 97% of cases of IDH (high to very high scores), a very high score was also found in 80% of cases of DCIS that were positive for CK-19. Of the 15 AIDHs, 80% had a negative or only focal reaction (negative or low score) for CK-34betaE12 and the remaining 20% had a moderate to high score for this antigen. Although CK-8 was strongly positive in 87% of cases of AIDH (high to very high scores), only 53.5% of AIDHs showed intense positivity for CK-19. The present study clearly shows that the immunoprofile of IDH is different from DCIS as far as HMW CK is concerned. Although florid IDH is characterized by a diffuse and intense immunoreaction for HMW CK, the lack of or only weak positivity for HMW CK (CK-34betaE12) is, in most cases, a hallmark of ductal carcinoma in situ. The immunoprofile of AIDH is very similar to that of DCIS. The expression of CK-8 and CK-19 is not useful in separating the various categories of ductal intraepithelial proliferations of the breast. We recommend the use of CK-34betaE12 as an adjunct in the diagnosis of a variety of problematic intraductal proliferations of the breast. PMID- 10478665 TI - Parachordoma is immunohistochemically and cytogenetically distinct from axial chordoma and extraskeletal myxoid chondrosarcoma. AB - Parachordoma is a rare soft-tissue tumor resembling extraskeletal myxoid chondrosarcoma and chordoma. Because fewer than 30 cases have been reported and precisely characterized, we studied the clinicopathologic features of seven new cases, comparing the immunohistochemical (six cases) and cytogenetic (one case) profiles with 15 cases of chordoma and six cases of extraskeletal myxoid chondrosarcoma. Patients with these tumors ranged in age from 7 to 62 years (mean, 35 years) and included four women and three men. The tumors presented as subfascial masses of the thigh (two cases), arm (three cases), chest wall (one cases), and buttocks (one case). In six of seven cases, there was neither recurrence nor metastasis within the follow-up, which ranged from 4 months to 7 years. The tumors were composed of vague nodules of large, rounded eosinophilic cells embedded in a matrix that varied from myxoid to densely hyaline, and the latter areas occasionally resembled primitive cartilage. Transitions between the large eosinophilic cells and smaller rounded and shorter spindled ones were often noted. Multivacuolated (physaliferouslike) cells were noted in all cases but were usually few in number. The matrix stained with Alcian blue (pH 2.5), and this staining was abolished with hyaluronidase predigestion. Immunohistochemistry for a variety of cytokeratins (CKs) (8/18, 1/10, 7, and 20), epithelial membrane antigen (EMA), S-100 protein, vimentin CD34, type IV collagen, smooth muscle actin, smooth muscle myosin heavy chain, calponin, and glial fibrillary acid protein was performed. All parachordomas strongly expressed CK 8/18, but not the other cytokeratins. Additionally, they expressed EMA (five of six). S-100 protein (six of six), and vimentin (six of six) and had a linear pattern of type IV collagen immunoreactivity around nests of cells (four of five). Calponin was noted in one case, but no cases expressed smooth muscle actin, smooth muscle myosin heavy chain, or glial fibrillary acid protein. In contrast, chordoma expressed CK 8/18 (15 of 15) and CK 1/10 (14 of 15), whereas extraskeletal myxoid chondrosarcoma consistently lacked CK. Although chordoma and extraskeletal myxoid chondrosarcoma showed considerable overlap with parachordoma, with respect to EMA and S-100 protein, they infrequently displayed type IV collagen, as was seen in parachordoma. One case of parachordoma studied cytogenetically disclosed trisomy 15, and monosomies of 1, 16, and 17 in contrast to the t(9;22) reported in extraskeletal myxoid chondrosarcoma and the monosomies of 3, 4, 10, and 13 seen in chordoma. We conclude that the immunohistochemical and cytogenetic profile distinguishes parachordoma from extraskeletal myxoid chondrosarcoma and chordoma. Lack of myoepithelial markers, furthermore, suggests parachordoma is not a deeply situated adnexal tumor. Because of these differences, parachordoma is best regarded as a distinct lesion without a clear relationship to other well characterized tumors. PMID- 10478667 TI - Focal lymphocytic colitis and collagenous colitis: patterns of Crohn's colitis? AB - The morphologic findings in mildly active colonic Crohn's disease (CD) include crypt disarray, patchy edema, and small lymphoid aggregates with neutrophils, sometimes associated with aphthous ulcers. We describe four patients with CD whose colonic biopsies focally showed a lymphocytic colitis morphology, and one patient with CD whose biopsies showed a collagenous colitis morphology. The lymphocytic and collagenous colitis patterns of injury preceded the eventual clinical pathologic diagnosis of CD in four patients. Colonoscopic abnormalities were found in four patients. The lymphocytic colitis pattern was focal, involving some biopsy fragments, whereas other biopsy fragments were normal or had minimal nonspecific inflammation. In one patient, moderate numbers of neutrophils were admixed with the lymphoplasmacytic infiltrates. The presence of colonoscopic abnormalities, focal changes, and moderate admixed neutrophils could assist in the distinction from lymphocytic or collagenous colitis, both of which are colonoscopically normal, usually diffuse, and devoid of, or contain only a sparse number of, neutrophils. A limited number of biopsy fragments may be incorrectly interpreted as lymphocytic or collagenous colitis. The temporal relationships suggest that these morphologic patterns precede typical active CD. PMID- 10478666 TI - Colonic epithelial lymphocytosis without a thickened subepithelial collagen table: a clinicopathologic study of 40 cases supporting a heterogeneous entity. AB - Lymphocytic colitis (LC) is classically described as a triad of chronic nonbloody, watery diarrhea, normal or nearly normal endoscopy findings, and colonic epithelial lymphocytosis without a thickened subepithelial collagen table (SECT). It is unknown how often patients with colonic epithelial lymphocytosis without a thickened SECT actually present with this classic triad. Cases diagnosed histologically as lymphocytic or microscopic colitis were reviewed. Criteria for inclusion were the presence of at least 15 surface lymphocytes per 100 epithelial cells and the absence of a thickened SECT (<12 microm). Clinical features and course were recorded by chart review and telephone follow-up. Forty patients met the inclusion criteria, including 25 women and 15 men with a mean age of 63.2 years (range, 25-83 years). Twenty-eight patients had the classic triad and were designated as having classic LC. The other 12 patients fulfilled the histologic criteria but not the clinical or endoscopic criteria for classic LC and were classified as having atypical LC (constipation, five patients; macroscopic colitis at endoscopy, five patients; hematochezia, one patient; and incidental finding, one patient). Clinically, patients with classic LC were predominantly women and had a higher incidence of autoimmune disease (p = 0.03) than did those with atypical LC. Histologically, surface eosinophilia was significantly greater in patients with classic LC (p = 0.04). Twenty patients were using nonsteroidal antiinflammatory drugs at the time of their colonic biopsy. Surface epithelial lymphocyte counts were higher in these patients, particularly in the distal sigmoid colon (p = 0.02). Fourteen patients had associated autoimmune disease, including three patients with sprue diagnosed by small bowel biopsy, all of whom responded to gluten withdrawal. Diarrhea present in 25 patients, without documented evidence of celiac sprue, was self-limited in five, resolved with treatment in three, required intermittent treatment in eight, daily treatment in five, and was refractory to treatment in four. All eight patients who experienced spontaneous or treatment-related symptom resolution had classic LC. No histologic feature correlated with clinical course. In conclusion, our study shows that colonic epithelial lymphocytosis without a thickened SECT is a histologic finding seen in a heterogeneous group of patients. Within this heterogeneous group is a distinct subset of patients who have the classic clinicopathologic triad of LC. This subset of patients has striking similarities to patients with collagenous colitis, lending further support to a close relationship between these two entities. Atypical LC comprises a heterogeneous group and includes patients with idiopathic constipation, coexisting LC and inflammatory bowel disease, and possibly infectious colitides. Because of the clinical heterogeneity among our study population, the descriptive term colonic epithelial lymphocytosis may be a more prudent diagnosis than lymphocytic colitis in the absence of adequate clinical information. PMID- 10478668 TI - Leiomyosarcoma of the pulmonary veins. AB - Primary sarcomas of the great vessels are rare, but the most common site is the inferior vena cava. Herein are reported five new cases arising from the pulmonary veins with clinicopathologic correlation and comparison to previously reported cases. All new cases occurred in women ranging in age from 23 to 64 years at diagnosis (mean, 56 years). They had symptoms suggestive of left heart failure, including three patients with dyspnea, one with hemoptysis, and one with cough. Three cases showed tumor extension along the pulmonary veins into the left atrium. Tumors ranged in size from 2.8 to 7 cm in greatest dimension. Histologically, all were leiomyosarcomas. They were highly cellular tumors. Three cases had predominantly spindle cell morphology and two were predominantly epithelioid; one had foci of calcification. Most showed extensive necrosis. All tumors were reactive with antibodies to actin and desmin. Two cases were reactive with antibodies to MIC-2 (dotlike); two cases showed reactivity to keratin antibodies; and two showed reactivity for estrogen, progesterone receptor protein, or both. None were positive for antibodies to S-100 protein. All cases were treated with surgical excision. Follow-up ranged from 2 months to 21 years (mean, 4.8 years). Two patients were alive and well; two were alive with metastases; and one died of disease. Pulmonary vein sarcomas represent intermediate- to high-grade leiomyosarcoma. Although often lethal, complete surgical excision can lead to long-term survival. They occur predominantly in women and may express hormone receptors. Therefore, hormonal manipulation may offer promise as adjuvant therapy. PMID- 10478669 TI - Adult granulosa cell tumor of the ovary with foci of hepatic cell differentiation: a report of four cases and comparison with two cases of granulosa cell tumor with Leydig cells. AB - We report four ovarian granulosa cell tumors of the adult type containing small foci of hepatic cell differentiation. The patients ranged in age from 35 to 54 years and had unilateral adnexal masses. The smallest tumor was 4.0 cm in diameter and the largest, 11.0 cm in diameter. Three tumors were solid and cystic, and one was cystic. Microscopic examination showed typical patterns of adult granulosa cell tumor, with the additional finding of scattered islands of large cells with abundant eosinophilic, slightly granular cytoplasm and central round nuclei containing single prominent nucleoli. Bile pigment was detected in canaliculi between some of the large cells in three tumors. The hepatic cells were positive immunohistochemically for cytokeratin (CAM 5.2) and epithelial membrane antigen in two cases and alpha-fetoprotein in one of two cases. Carcinoembryonic antigen was stained in a canalicular pattern in two cases. Staining for vimentin and alpha-inhibin was negative. Liver cells in granulosa cell tumors must be differentiated from Leydig cells, which are found very rarely in granulosa cell tumors, and luteinized stromal and granulosa cells, which are present more commonly in these tumors; all three of the latter cell types are positive for alpha-inhibin. PMID- 10478670 TI - Renal oncocytosis: a morphologic study of fourteen cases. AB - Diffuse renal involvement by numerous oncocytic nodules has rarely been described. We report 14 cases (19 specimens) with innumerable oncocytic nodules in the kidney. Invariably, these kidneys showed additional associated findings. We suggest the term renal oncocytosis for this entire morphologic spectrum. Six (43%) cases had histologically or radiologically proven bilateral involvement. Each specimen had at least one dominant tumor (2.0-10.5 cm) in addition to numerous other microscopic to macroscopic oncocytic nodules. Additional features observed were: interstitial pattern, with the oncocytic tubules and acini diffusely intermingling with and infiltrating between non-neoplastic parenchyma (one case); diffuse oncocytic change in the nonneoplastic tubules, cytologically difficult to separate from the oncocytic nodules (seven cases); and benign oncocytic cortical cysts (four cases). The dominant mass in 13 specimens was a renal oncocytoma and in two, a chromophobe renal cell carcinoma. In four specimens, the largest tumor was considered a hybrid tumor because of the presence of mixed histologic features of both tumor types. Most smaller nodules had the morphologic features of renal oncocytoma, but a few had the appearance of chromophobe renal cell carcinoma or nodules with hybrid features. We conclude that the presence of numerous oncocytic nodules may be associated with a wide spectrum of oncocytic changes in the kidney. The association of numerous renal oncocytoma-like nodules with lesions having a mixed morphology or a morphology of pure chromophobe renal cell carcinoma suggests that they may constitute a morphologic spectrum of oncocytic tumors and that renal oncocytoma and chromophobe renal cell carcinoma may arise from a common progenitor lesion. PMID- 10478671 TI - A multicentric European study testing the reproducibility of the WHO classification of endometrial hyperplasia with a proposal of a simplified working classification for biopsy and curettage specimens. AB - This study was designed to assess intraobserver and interobserver agreement in the diagnosis of 56 endometrial specimens by five European expert gynecologic pathologists using the WHO classification and to establish which histologic features are significantly associated with each classification category. The seven categories were simple hyperplasia, complex hyperplasia, atypical hyperplasia, well-differentiated adenocarcinoma, proliferative endometria, secretory endometria, and other. Slides were reviewed twice for diagnosis, with accompanying evaluation of a checklist of histologic features. These seven categories were eventually reduced to four and three for the purposes of data analysis. The four modified diagnostic categories consisted of hyperplasia (previously simple hyperplasia and complex hyperplasia), atypical hyperplasia, well-differentiated adenocarcinoma, and cyclical endometrium (previously proliferative, secretory, and other). The three diagnostic categories consisted of hyperplasia, endometrioid neoplasia (previously atypical hyperplasia and well differentiated adenocarcinoma), and cyclical endometrium. Intraobserver and interobserver agreement was assessed using the percentage agreement and kappa statistics. The associations among the various histologic features and diagnoses was analyzed using multiple logistic regression to identify those features that were useful for distinguishing diagnostic categories. When using seven categories, kappa values ranged from 0.53 to 0.74 (percentage agreement, 61-79%) and from 0.33 to 0.59 (percentage agreement, 43-63%) for intraobserver and interobserver agreement, respectively. When using four categories, kappa values ranged from 0.68 to 0.73 (percentage agreement, 77-80%) and from 0.39 to 0.64 (percentage agreement, 54-73%) for intraobserver and interobserver agreement, respectively. When using three categories, kappa values ranged from 0.70 to 0.83 (percentage agreement, 80-89%) and from 0.55 to 0.73 (percentage agreement, 70 82%) for intraobserver and interobserver agreement, respectively. Data were analyzed in each diagnostic category. When using four or three diagnostic categories, the mean intraobserver and interobserver agreements varied less between categories and achieved higher values, with smaller 95% confidence intervals. The mean percentage agreement was lowest for complex hyperplasia and for atypical hyperplasia. For distinguishing cyclical endometrium versus hyperplasia, the useful histologic feature was glandular crowding. For hyperplasia versus atypical hyperplasia and for hyperplasia versus endometrioid neoplasia, the useful features were nuclear enlargement, nuclear pleomorphism, vesicular chromatin, and nucleoli, but of these, only nuclear pleomorphism achieved substantial mean intraobserver and interobserver agreements. For discriminating atypical hyperplasia from well-differentiated adenocarcinoma, the only useful feature was stromal alterations, which achieved only fair mean intraobserver and interobserver agreements. In summary, in endometrial biopsy or curettage specimens, the lack of agreement in the diagnoses of complex hyperplasia and atypical hyperplasia and the lack of reproducibility in the recognition of the histologic feature of stromal alterations to differentiate atypical hyperplasia from well-differentiated adenocarcinoma suggest that the histologic classification should be simplified by including a combined category for simple and complex hyperplasia, called hyperplasia, and a combined category for atypical hyperplasia and well-differentiated adenocarcinoma, called endometrioid neoplasia. Diagnoses of hyperplasia and endometrioid neoplasia are highly reproducible between observers from different institutions. Glandular crowding is the best histologic feature to differentiate cyclical endometrium from hyperplasia, whereas nuclear pleomorphism is the reproducible cytologic feature to differentiate hyperplasia from endometrioid neoplasia. PMID- 10478672 TI - Gastrointestinal stromal tumors/smooth muscle tumors (GISTs) primary in the omentum and mesentery: clinicopathologic and immunohistochemical study of 26 cases. AB - Gastrointestinal stromal tumor or smooth muscle tumor (GIST) is the designation for a major subset of gastrointestinal mesenchymal tumors that histologically, immunohistochemically, and genetically differ from typical leiomyomas, leiomyosarcomas, and schwannomas. Because GISTs, like the interstitial cells of Cajal, the gastrointestinal pacemaker cells, express CD117 (c-kit protein), the origin of GISTs from the interstitial cells of Cajal has been recently proposed. Comparison of GISTs primary in the omentum and mesentery to GISTs primary in the tubular gastrointestinal tract is of particular diagnostic and histogenetic interest in view of the possible similarity of these tumors with the GIST group. In this study, we analyzed 14 omental and 12 mesenteric primary mesenchymal tumors representing smooth muscle tumors or GISTs. These tumors were phenotypically compared with gastric and small intestinal GISTs, leiomyomas of the esophagus, and leiomyosarcomas of the retroperitoneum. Most (13 of 14) omental and mesenteric (10 of 12) tumors showed histologic features similar to GISTs with elongated spindle cells or epithelioid cells with high cellularity; most of these tumors showed low mitotic activity. Omental and mesenteric GISTs were typically positive for CD117 and less consistently for CD34. They often showed alpha-smooth muscle actin reactivity but were virtually negative for desmin and S-100 protein. One omental and two mesenteric tumors showed features of leiomyosarcoma with ovoid, less elongated nuclei, cytoplasmic eosinophilia; all these tumors had significant mitotic activity. These tumors were positive for alpha-smooth muscle actin and two of them for desmin, but all were negative for CD34 and CD117, similar to retroperitoneal leiomyosarcomas. Tumor-related mortality occurred in the group of mesenteric GISTs, but not in the group of omental GISTs. In contrast, all three patients with a true leiomyosarcoma of the omentum or mesentery had documented liver metastases or died of tumor. In summary, we show that tumors phenotypically identical with GISTs occur as primary tumors in the omentum and mesentery. The occurrence of CD117-positive tumors outside the gastrointestinal tract militates against an origin of these tumors exclusively from the interstitial cells of Cajal. PMID- 10478673 TI - Human papillomavirus-associated penile squamous cell carcinoma in HIV-positive patients. AB - Two cases of penile squamous cell carcinoma with distinctive clinicopathologic characteristics are presented. The tumors appeared in patients infected with HIV and were located in the glans of the penis. Histologically, the neoplasms were well-differentiated, infiltrating, squamous cell carcinomas. The entire spectrum from benign condyloma to infiltrative squamous cell carcinoma was present in the two patients. In both cases, human papillomavirus (HPV) could be demonstrated using polymerase chain reaction analysis. The reported cases suggest a synergic interaction of HPV and HIV in the carcinogenic process of some penile carcinomas. PMID- 10478674 TI - Primary hepatic diffuse large B-cell lymphoma in a patient with chronic hepatitis C. AB - Epidemiological and experimental data suggest that the hepatitis C virus infection might be associated with the development of distinct types of non Hodgkin's lymphomas. Here, we report a case of a patient with chronic hepatitis C and type II mixed cryoglobulinemia, who developed a primary hepatic non-Hodgkin's B-cell lymphoma. A diffuse, large B-cell lymphoma was diagnosed based on morphological, immunophenotypical and molecular genetic findings. Hepatitis C virus replication, as evaluated by strand-specific reverse transcriptase polymerase chain reaction, was detected in the nonneoplastic liver, but not in the lymphomatous tissue. High grade non-Hodgkin's lymphomas, although rare complications, have to be considered as part of the spectrum of hepatitis C virus related hepatic lesions. PMID- 10478675 TI - Microcystic meningioma arising in a mixed germ cell tumor of the testis: a case report. AB - We report a case of a microcystic variant of meningioma arising in a mixed germ cell tumor of the testis composed predominantly of mature and immature teratoma with elements of seminoma and embryonal carcinoma. We believe this is the first such case of a meningioma arising in a teratoma within a gonadal or extragonadal site. The meningiomatous component showed positive immunohistochemical staining for epithelial membrane antigen and a lack of staining for cytokeratin, factor VIII, CD31, and alpha-fetoprotein. Recognition of a non-germ cell tumor arising in the setting of a teratoma in the testis may be prognostically important depending on the nature of the non-germ cell component and whether it has spread beyond the testis. PMID- 10478676 TI - Pigmented central neurocytoma: case report and literature review. AB - A case of pigmented central neurocytoma is reported. The tumor showed histologic, immunophenotypic, and ultrastructural features of central neurocytoma. The pigment consisted of an intimate association of lipofuscin and neuromelanin. Pigmented neuroepithelial tumors are rare, and the pigments may be neuromelanin or melanin. Pigmented central neurocytoma has not been described to date. The low proliferation rate of the tumor suggests a delayed turnover of tumor cells as a possible cause of lipofuscin accumulation. Autocatalytic peroxidation of lipofuscin is a possible mechanism of neuromelanin formation. PMID- 10478677 TI - Interdigitating dendritic cell tumor of the testis: a novel testicular spindle cell neoplasm. AB - Interdigitating dendritic cell tumor is an extremely rare neoplasm that mainly occurs in lymph nodes. An example of such a tumor in the testis, a hitherto unreported site, is described. Grossly, the tumor was light tan with a uniform solid appearance, replacing virtually the entire testis. Microscopically, it was formed by whorls and fascicles of spindle cells intermingling with small lymphocytes. Such a histologic appearance can, however, mimic a wide variety of other tumors and tumor-like lesions, among which mesenchymal sarcoma, spindle cell carcinoma, follicular dendritic cell tumor, and inflammatory pseudotumor are the main differential diagnoses. Immunohistochemical studies showed that the spindle tumor cells were strongly and diffusely positive for S-100 protein and vimentin. They were also focally positive for CD68 and CD4, but were uniformly negative for leukocyte common antigen, CD1a, CD3, CD20, CD21, CD23, CD34, CD35, actin, desmin, HMB45, cytokeratins, and placental alkaline phosphatase. Ultrastructurally, the tumor cells possessed complex interdigitating cytoplasmic dendritic processes, with abundant rough endoplasmic reticulum and mitochondria in their cytoplasm. An in situ hybridization study for Epstein-Barr virus was negative. The pathologist should be aware of such an entity and consider it in the list of differential diagnoses for unusual spindle cell lesions with a significant background population of small lymphocytes. However, because of its nonspecific histologic appearance, additional immunohistochemical and electron microscopic studies are generally required for its definitive diagnosis. PMID- 10478678 TI - Colonic primary large cell lymphoma with marginal zone growth pattern presenting as multiple polyps. AB - We report a rare case of primary gastrointestinal lymphoma, stage IE, in a 58 year-old white man who had multiple colonic polyps measuring up to 1 x 1.1 cm. The tumor originated in the marginal zone of the follicles infiltrating the interfollicular spaces. Follicular colonization was frequently seen. The mucosa was spared by the infiltrate. Morphologically, the neoplastic cells were monomorph, intermediate-sized blasts. Rare small to intermediate sized cells, some with centrocyte-like morphology, intermingled the blastic infiltrate. The neoplastic cells expressed CD20 and had a monotypic immunoglobulin of cytoplasmic IgM (kappa) on paraffin sections. Tumor cells stained negative for CD45RO, CD5, CD10, IgD, and CD23. Polymerase chain reaction revealed a clonal V-D-J rearrangement. Bcl-1 and bcl-2 rearrangement were not detected. We therefore suggest the diagnosis of primary large cell lymphoma with marginal zone growth pattern mimicking colonic adenomas. PMID- 10478679 TI - Neurofibroma and cellular neurofibroma with atypia: a report of 14 tumors. PMID- 10478680 TI - Small hyperplastic polyps of the colorectum showing deranged cell organization: a lesion considered to be a serrated adenoma? PMID- 10478681 TI - Immature ovarian teratomas in children. PMID- 10478682 TI - Lymphomatoid granulomatosis. PMID- 10478684 TI - What should I do if my patient does not speak English? PMID- 10478683 TI - The patient-physician relationship: is three a crowd? PMID- 10478685 TI - Lessons outside the lecture hall. PMID- 10478686 TI - Unlicensed vaccines and bioweapon defense in World War II. PMID- 10478687 TI - Tobacco dependence curricula in US undergraduate medical education. AB - CONTEXT: Tobacco use is the leading preventable cause of death in the United States. And yet only 21% of practicing physicians claim they received adequate training to help their patients stop smoking. OBJECTIVE: To assess the content and extent of tobacco education and intervention skills in US medical schools' curricula. DESIGN: A survey with 13 multiple-response items on tobacco education. Survey questions were based on the recommendations of the Agency for Health Care Policy and Research and the National Cancer Institute Expert Panel. The Liaison Committee on Medical Education included 4 of these items in a modified form on the 1997 annual questionnaire. SETTING: One hundred twenty-six US medical schools. PARTICIPANTS: Surveys were obtained from 122 associate deans for medical education (98.6%). MAIN OUTCOME MEASURES: Curriculum content in basic science and clinical science, elective or required clinical experience, hours of instruction, and resource materials. RESULTS: Inclusion of all 6 tobacco curricula content areas recommended by the National Cancer Institute and the Agency for Health Care Policy and Research was higher in basic science (63/115 [54.8%]) than in clinical science (5/115 [4.4%]). Most medical schools (83/120 [69.2%]) did not require clinical training in smoking cessation techniques, while 23.5% (27/115) offered additional experience as an elective course. Thirty-one percent (32/102) of schools averaged less than 1 hour of instruction per year in smoking cessation techniques during the 4 years of medical school. A minority of schools reported 3 or more hours of clinical smoking cessation instruction in the third (14.7%) and fourth (4.9%) years. CONCLUSIONS: A majority of US medical school graduates are not adequately trained to treat nicotine dependence. The major deficit is the lack of smoking cessation instruction and evaluation in the clinical years. A model core tobacco curricula that meets national recommendations should be developed and implemented in all US medical schools. PMID- 10478688 TI - Teaching professionalism in undergraduate medical education. AB - CONTEXT: There is a growing consensus among medical educators that to promote the professional development of medical students, schools of medicine should provide explicit learning experiences in professionalism. OBJECTIVE: To determine whether and how schools of medicine were teaching professionalism in the 1998-1999 academic year. DESIGN, SETTING, AND PARTICIPANTS: A 2-stage survey was sent to 125 US medical schools in the fall of 1998. A total of 116 (92.3%) responded to the first stage of the survey. The second survey led to a qualitative analysis of curriculum materials submitted by 41 schools. MAIN OUTCOME MEASURES: Presence or absence of learning experiences (didactic or experiential) in undergraduate medical curriculum explicitly intended to promote professionalism in medical students, with curriculum evaluation based on 4 attributes commonly recognized as essential to professionalism: subordination of one's self-interests, adherence to high ethical and moral standards, response to societal needs, and demonstration of evincible core humanistic values. RESULTS: Of the 116 responding medical schools, 104 (89.7%) reported that they offer some formal instruction related to professionalism. Fewer schools have explicit methods for assessing professional behaviors (n = 64 [55.2%]) or conduct targeted faculty development programs (n = 39 [33.6%]). Schools use diverse strategies to promote professionalism, ranging from an isolated white-coat ceremony or other orientation experience (n = 71 [78.9%]) to an integrated sequence of courses over multiple years of the curriculum (n = 25 [27.8%]). Of the 41 schools that provided curriculum materials, 27 (65.9%) addressed subordinating self-interests; 31 (75.6%), adhering to high ethical and moral standards; 17 (41.5%), responding to societal needs; and 22 (53.7%), evincing core humanistic values. CONCLUSIONS: Our results suggest that the teaching of professionalism in undergraduate medical education varies widely. Although most medical schools in the United States now address this important topic in some manner, the strategies used to teach professionalism may not always be adequate. PMID- 10478689 TI - Mindful practice. AB - Mindful practitioners attend in a nonjudgmental way to their own physical and mental processes during ordinary, everyday tasks. This critical self-reflection enables physicians to listen attentively to patients' distress, recognize their own errors, refine their technical skills, make evidence-based decisions, and clarify their values so that they can act with compassion, technical competence, presence, and insight. Mindfulness informs all types of professionally relevant knowledge, including propositional facts, personal experiences, processes, and know-how, each of which may be tacit or explicit. Explicit knowledge is readily taught, accessible to awareness, quantifiable and easily translated into evidence based guidelines. Tacit knowledge is usually learned during observation and practice, includes prior experiences, theories-in-action, and deeply held values, and is usually applied more inductively. Mindful practitioners use a variety of means to enhance their ability to engage in moment-to-moment self-monitoring, bring to consciousness their tacit personal knowledge and deeply held values, use peripheral vision and subsidiary awareness to become aware of new information and perspectives, and adopt curiosity in both ordinary and novel situations. In contrast, mindlessness may account for some deviations from professionalism and errors in judgment and technique. Although mindfulness cannot be taught explicitly, it can be modeled by mentors and cultivated in learners. As a link between relationship-centered care and evidence-based medicine, mindfulness should be considered a characteristic of good clinical practice. PMID- 10478690 TI - Educational programs in US medical schools, 1998-1999. AB - To describe the current status of medical education programs in the United States and to trace trends in medical education over this century, we used data from the 1998-1999 Liaison Committee on Medical Education Annual Medical School Questionnaire, which had a 100% response rate, and data from other sources. In 1998-1999, total full-time faculty members numbered 98202, a 1.5% increase from 1997-1998. The number of applicants to medical school declined for the second consecutive year, from 43020 in 1997 to 41004 in 1998, but the academic qualifications of entering students remained steady. The number of applicants from underrepresented minority groups decreased 1.3% from 1997 to 1998, compared with an 11.1% decrease between 1996 and 1997. Women constituted 43.4% of applicants in 1998, slightly more than the 42.5% in 1997. The total number of required hours in the first and second years of the curriculum and the number of scheduled hours per week have declined over the past 15 years, while the average lengths of clinical clerkships remained about the same. The number of schools requiring students to pass Steps 1 and 2 of the United States Medical Licensing Examination continued to increase in 1998-1999, with 50% of schools requiring passing both examinations, compared with 46% in 1997-1998. PMID- 10478691 TI - Review of US medical school finances, 1997-1998. AB - Based on data from the Annual Medical School Questionnaire of the Liaison Committee on Medical Education (LCME), to which 100% of schools responded, we found that revenue supporting programs and activities of the 125 accredited medical schools in the United States totaled $36997 million in 1997-1998. A large proportion of revenue (79%) was derived from 3 sources: practice plans ($12559 million; 33.9%), grants and contracts ($10916 million; 29.5%), and hospital support ($5741 million; 15.5%). An analysis of revenue trends revealed that medical schools, in aggregate, have continued to experience growth during the last 2 years. However, the aggregate numbers mask considerable variation among schools with regard to changes in financing. Between 1995-1996 and 1996-1997, 46 schools (37%) reported constant-dollar declines in the sum of practice plan and hospital revenue, and 50 schools (40%) reported a decline from 1996-1997 to 1997 1998. The financial data reviewed in this report demonstrate the continued dependence of medical schools on faculty-generated sources of revenue and confirm the perception that a growing number of medical schools are experiencing reductions in key sources of financial support. Current and projected reductions in teaching hospital revenue due to the implementation of the Balanced Budget Amendment are expected to erode further hospital support for medical school programs and activities. PMID- 10478692 TI - Graduate medical education, 1998-1999: a closer look. AB - Despite the ever-present risk of a critical imbalance in the physician workforce total numbers and specialty distribution, no systematic planning mechanism is in place. Furthermore, the length of training for graduate medical education (GME) precludes timely rectification of imbalances. We report GME activities collected in the American Medical Association Annual Survey of Graduate Medical Education Programs for 1998-1999, along with trends during the last 3 to 6 years. These data initially suggest that little has changed during the past several years; however, on closer examination, small but significant changes during the past 2 years may have serious consequences if continued. The total number of resident physicians, which has been constant during the last several years, decreased in 1998 by 760 from the previous year, while the number of programs continues to increase (6.1% since 1993). The number of US medical graduates entering GME programs remained stable during the last 6 years, the number of osteopathic graduates entering GME increased by 55.7%, and the number of international medical graduates (IMGs) entering GME continued to drop (down 13.2% since 1993). More IMGs tend to pursue additional training than do US graduates (in 1997, 32.9% vs 23.6%). Because IMGs are remaining in GME programs for longer periods, the total number of IMGs has not yet reflected significant change. About 62% of IMGs now entering GME training are either US citizens or permanent residents; ethnic minority residents are not decreasing in numbers as some predicted; and for the first time in the past 5 years, the primary care specialties have ceased their persistent growth. Toward the end of GME, the number of residents leaving programs before completion increased by 5.7% during the last 3 years. While some of these changes may be ascribed to reduced GME funding through the Balanced Budget Act of 1997, other factors clearly are at play. To anticipate future changes in the physician workforce, these factors should be identified to permit them to be monitored and modified as needed. PMID- 10478693 TI - Simulation technology for health care professional skills training and assessment. AB - Changes in medical practice that limit instruction time and patient availability, the expanding options for diagnosis and management, and advances in technology are contributing to greater use of simulation technology in medical education. Four areas of high-technology simulations currently being used are laparoscopic techniques, which provide surgeons with an opportunity to enhance their motor skills without risk to patients; a cardiovascular disease simulator, which can be used to simulate cardiac conditions; multimedia computer systems, which includes patient-centered, case-based programs that constitute a generalist curriculum in cardiology; and anesthesia simulators, which have controlled responses that vary according to numerous possible scenarios. Some benefits of simulation technology include improvements in certain surgical technical skills, in cardiovascular examination skills, and in acquisition and retention of knowledge compared with traditional lectures. These systems help to address the problem of poor skills training and proficiency and may provide a method for physicians to become self directed lifelong learners. PMID- 10478694 TI - Impact of formal continuing medical education: do conferences, workshops, rounds, and other traditional continuing education activities change physician behavior or health care outcomes? AB - CONTEXT: Although physicians report spending a considerable amount of time in continuing medical education (CME) activities, studies have shown a sizable difference between real and ideal performance, suggesting a lack of effect of formal CME. OBJECTIVE: To review, collate, and interpret the effect of formal CME interventions on physician performance and health care outcomes. DATA SOURCES: Sources included searches of the complete Research and Development Resource Base in Continuing Medical Education and the Specialised Register of the Cochrane Effective Practice and Organisation of Care Group, supplemented by searches of MEDLINE from 1993 to January 1999. STUDY SELECTION: Studies were included in the analyses if they were randomized controlled trials of formal didactic and/or interactive CME interventions (conferences, courses, rounds, meetings, symposia, lectures, and other formats) in which at least 50% of the participants were practicing physicians. Fourteen of 64 studies identified met these criteria and were included in the analyses. Articles were reviewed independently by 3 of the authors. DATA EXTRACTION: Determinations were made about the nature of the CME intervention (didactic, interactive, or mixed), its occurrence as a 1-time or sequenced event, and other information about its educational content and format. Two of 3 reviewers independently applied all inclusion/exclusion criteria. Data were then subjected to meta-analytic techniques. DATA SYNTHESIS: The 14 studies generated 17 interventions fitting our criteria. Nine generated positive changes in professional practice, and 3 of 4 interventions altered health care outcomes in 1 or more measures. In 7 studies, sufficient data were available for effect sizes to be calculated; overall, no significant effect of these educational methods was detected (standardized effect size, 0.34; 95% confidence interval [CI], -0.22 to 0.97). However, interactive and mixed educational sessions were associated with a significant effect on practice (standardized effect size, 0.67; 95% CI, 0.01-1.45). CONCLUSIONS: Our data show some evidence that interactive CME sessions that enhance participant activity and provide the opportunity to practice skills can effect change in professional practice and, on occasion, health care outcomes. Based on a small number of well-conducted trials, didactic sessions do not appear to be effective in changing physician performance. PMID- 10478695 TI - Educating medical students for work in culturally diverse societies. AB - CONTEXT: Recent attention has focused on whether government health service institutions, particularly in the United Kingdom, reflect cultural sensitivity and competence and whether medical students receive proper guidance in this area. OBJECTIVE: To systematically identify educational programs for medical students on cultural diversity, in particular, racial and ethnic diversity. DATA SOURCES: The following databases were searched: MEDLINE (1963-August 1998); Bath International Data Service (BIDS) Institute for Scientific Information science and social science citation indexes (1981-August 1998); BIDS International Bibliography for the Social Sciences (1981-August 1998); and the Educational Resources Information Centre (1981-August 1998). In addition, the following online data sets were searched: Kings Fund; Centre for Ethnic Relations, University of Warwick; Health Education Authority; European Research Centre on Migration and Ethnic Relations, University of Utrecht; International Centre for Intercultural Studies, University of London; the Refugee Studies Programme, University of Oxford. Medical education and academic medicine journals (1994 1998) were searched manually and experts in medical education were contacted. STUDY SELECTION: Studies included in the analysis were articles published in English before August 1998 that described specific programs for medical students on racial and ethnic diversity. Of 1456 studies identified by the literature search, 17 met the criteria. Two of the authors performed the study selection independently. DATA EXTRACTION: The following data were extracted: publication year, program setting, student year, whether a program was required or optional, the teaching staff and involvement of minority racial and ethnic communities, program length, content and teaching methods, student assessment, and nature of program evaluation. DATA SYNTHESIS: Of the 17 selected programs, 13 were conducted in North America. Eleven programs were exclusively for students in years 1 or 2. Fewer than half (n = 7) the programs were part of core teaching. Only 1 required program reported that the students were assessed on the session in cultural diversity. CONCLUSIONS: Our study suggests that there is limited information available on an increasingly important subject in medical education. Further research is needed to identify effective components of educational programs on cultural diversity and valid methods of student assessment and program evaluation. PMID- 10478696 TI - Instilling professionalism in medical education. PMID- 10478697 TI - JAMA Patient Page. Tobacco. PMID- 10478698 TI - New directions in the management of hypertension and the cardiovascular risk. Introduction. PMID- 10478699 TI - Overview of JNC VI: new directions in the management of hypertension and cardiovascular risk. AB - Treatment recommendations for hypertension as outlined in the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI) are constantly evolving and being refined as new information on the disease becomes evident. Uncontrolled hypertension is a major antecedent of stroke, heart failure, coronary heart disease, and end-stage renal disease. The increasing incidences of both cardiovascular and renal diseases fuel the need for improved control of hypertension. In fact, according to the National Health and Nutrition Examination Survey (NHANES), about 69% of Americans whose blood pressure is greater than 140/90 mm Hg are aware of it, about half are getting treatment for it, and only about one-quarter are adequately controlled. These observations fuel the need for improved patient management guidelines. JNC VI makes several changes from the previous JNC V to assist physicians in the diagnosis, treatment, and improved management of patients with hypertension. These changes include reporting adult blood pressure in two new ways, via staging and risk factor classification. A high-normal classification (systolic: 130 to 139 mm Hg, or diastolic: 85 to 89 mm Hg) is included in JNC VI because of the clinical importance of such blood pressure contributing to cardiovascular disease. Additionally, clinicians are advised to assign a patient to one of three risk categories that, in addition to hypertension stage, influence the decision to select antihypertensive drug therapy. Lifestyle modification is an important component at each stage. These and other changes and highlights of recent studies supporting the need for more intensive blood pressure control are discussed in this paper. PMID- 10478700 TI - Low-dose antihypertensive combination therapy: its rationale and role in cardiovascular risk management. AB - Antihypertensive monotherapy, although commonly used, does not address the multifactorial nature of hypertension as a disease with many pathways. Using more than one drug makes more therapeutic sense because combination agents cover more than one pathway, yet the use of drugs in tandem is typically relegated to more problematic patients later in therapy. Many patients with hypertension are not controlled, because the monotherapeutic agent is used at its highest dose, resulting in side effects that lead to noncompliance. As opposed to fixed-dose combinations that merge two drugs at their highest doses, low-dose combination therapy provides more novel coverage of two or more metabolic pathways that contribute to hypertension. Their once-daily dosing encourages compliance. In addition, because the two drugs are combined at low doses, the probability of side effects is decreased and efficacy is often enhanced. The use of low-dose combination antihypertensive agents is a good contemporary strategy for first line therapy in that patients can take advantage of their cardiovascular benefits and the control these agents offer early in therapy. PMID- 10478701 TI - Benefits of combination angiotensin-converting enzyme inhibitor and calcium antagonist therapy for diabetic patients. AB - Effective control of blood pressure is important in slowing renal disease progression in diabetic nephropathy. However, blood pressure is often difficult to control with monotherapy, especially in this patient population in whom three medications are sometimes required to attain goal blood pressures of less than 130/85 mm. Further, physicians are often reluctant to increase the dose or add another agent to an existing regimen. The advent of low-dose, fixed-dose combination therapy allows physicians the ability to prescribe two drugs in one pill and, hence, to improve compliance. In fact, the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI) recommends adding a second antihypertensive agent as an option if goal blood pressure is not achieved with a single agent. The combination of an angiotensin-converting enzyme (ACE) inhibitor and calcium antagonist may confer additive antihypertensive benefit, and may even improve the side-effect profile. This paper reviews the importance of aggressive blood pressure control in diabetic patients and the clinical trials supporting the use of combination ACE inhibitor/calcium antagonist therapy in the diabetic patient population. PMID- 10478703 TI - Animal models of nonconvulsive status epilepticus. AB - Nonconvulsive status epilepticus includes three clinical situations: complex partial status epilepticus; absence status epilepticus: and obtundation in the presence of electrographic status epilepticus. Animal models that provide information helpful to clinical management exist for both complex partial and absence status epilepticus. In models of complex partial status epilepticus (pilocarpine, kainic acid, and various protocols using electrical stimulation), neuronal damage in discrete neuronal populations follows an episode of status epilepticus. Hippocampal populations are particularly susceptible to neuropathologic sequelae. Although it is difficult in some cases to distinguish whether the inducing agent or the status epilepticus causes neuropathology, the similar patterns of damage caused by different inducing stimuli provide converging lines of evidence suggesting that the neuropathologic consequences stem at least in part from status epilepticus. In models of absence status epilepticus (genetic mutants, pentylenetetrazole), there is relatively scarce neuropathology that can be attributed directly to status epilepticus. Together these data from animal models suggest that neuropathologic consequences from complex partial status epilepticus may be more severe than those from absence status epilepticus. If these findings translate to patients, then nonconvulsive status epilepticus of the complex partial type should be managed more aggressively than nonconvulsive status epilepticus of the absence type. PMID- 10478702 TI - Combinations in the treatment of hypertension: ACE inhibitors and calcium antagonists. AB - Hypertension control and management of concomitant pathophysiologic conditions may require use of multiple drugs. However, most studies in hypertensive disease have focused on monotherapy. Therefore, our knowledge of combination therapy in the treatment of hypertension is largely extrapolated from these monotherapy studies. Angiotensin-converting enzyme (ACE) inhibitors and calcium antagonist combinations should be particularly efficacious in reducing hypertensive target organ disease. Both of these drug classes have been shown to reduce complications of hypertensive heart disease and renal disease progression. With regard to hypertensive vascular disease, both ACE inhibitors and calcium antagonists have documented benefits. However, their use together in left ventricular hypertrophy and in patients with coronary heart disease, although promising, must be proved through carefully designed, prospective, randomized trials. PMID- 10478704 TI - Epidemiology and evidence for morbidity of nonconvulsive status epilepticus. AB - Convulsive status epilepticus (SE) is convincingly related to serious morbidity and mortality and well recognized as a medical emergency, but prompt diagnosis and treatment of patients with nonconvulsive status epilepticus (NCSE) is often not emphasized because its consequences are thought to be benign. Nonconvulsive status epilepticus has been considered a relatively benign entity because it does not produce the adverse systemic consequences of convulsive status epilepticus, such as hyperthermia, acidosis, hyperkalemia, pulmonary compromise, or cardiovascular collapse. However, recent reports indicate that NCSE is not so benign. There are two major forms of NCSE, absence status epilepticus and complex partial status epilepticus. Typical absence status epilepticus does not appear to have very serious consequences and may be a type of "inhibitory" seizure, but complex partial status epilepticus has been associated with serious morbidity and mortality. Despite not causing the systemic physiologic or metabolic derangements seen with convulsive SE, complex partial status epilepticus is still associated with the two other major factors correlated with poor outcomes in convulsive SE: 1) neuronal damage from abnormal electrical activity and 2) the interaction of acute neurologic disorders, such as stroke, that may precipitate SE. Other similar epileptiform encephalopathies such as "subclinical," "electroencephalographic," "nontonic-clonic," and "subtle" SE have not been as well studied as NCSE but pose similar issues. Early diagnosis and aggressive intervention have proven the best means of averting adverse outcomes in patients with convulsive SE. The diagnosis and treatment of NCSE, particularly complex partial status epilepticus, merit similar emphasis and attention. PMID- 10478705 TI - Evidence against permanent neurologic damage from nonconvulsive status epilepticus. AB - Nonconvulsive status epilepticus (NCSE) is much more common than is generally appreciated and is certainly underdiagnosed, but its long-term effects are largely undetermined and remain controversial. There is increasing experimental evidence that generalized convulsive status epilepticus produces lasting neuropathologic damage in the hippocampus, but experimental models often include provocation of status epilepticus (SE) by physical (e.g., electrical stimulation) and chemical (including excitotoxic) agents that may induce damage independent of the epileptiform discharges. Also, damage appears to be related to the intensity and duration of electrical stimulation. Such models usually include high frequency discharges sustained over long periods, somewhat different from the electrical activity of typical human NCSE. Pathologic studies in humans pertain primarily to patients who have had generalized convulsive status epilepticus. Clinical studies of the effects of NCSE are mandatory, but conclusions are difficult to come by, in part because of diverse definitions of NCSE. An altered mental status is obligatory, but the pertinent EEG and medication response criteria are controversial. Response to medication can be delayed by many hours or even days. Absence SE appears to cause no lasting effects. Complex partial SE is less uniform. Most reported cases have returned to baseline neurologic function, but several well-described patients have had prolonged memory deficits. The significance of other deficits is difficult to interpret in light of concomitant vascular and other diseases causing neurologic dysfunction. Clinical series usually lack premorbid neurologic and neuropsychologic assessment. The few exceptions are complicated by preexisting mental retardation and other deficits, by the coexistence of progressive illness, by the later effects of recurrent seizures, and almost always by the confounding influence of anticonvulsant medications. Most morbidity appears attributable to the underlying illnesses rather than to the NCSE itself. It is possible that relatively infrequent cases of prolonged NCSE or those with the synergistic effect of concomitant systemic illness, focal lesions, or very rapid excitatory epileptiform discharges may suffer more long-lasting damage, but these observations are still preliminary. NCSE should be treated expeditiously because of the acute neurologic impairment of the patients, because of the attendant morbidity including physical injury, and because it may go on to generalized convulsions. There is reasonable concern about possible long-term effects, but permanent neurologic damage from NCSE has not yet been established as a mandate for urgent treatment. PMID- 10478706 TI - Nonconvulsive status epilepticus in acute brain injury. AB - Whether or not nonconvulsive status epilepticus produces permanent brain damage is a source of controversy. Contributing to the controversy is the lack of clarity for classifying the clinical and electrographic phenomena that constitute nonconvulsive status epilepticus. Nonconvulsive status epilepticus commonly occurs in the context of an acute brain injury. For example, it commonly persists in generalized convulsive status epilepticus after convulsive activity has stopped, and it is not uncommonly associated with acute cerebral ischemia. Its clinical characteristics are ambiguous, subtle, and nonspecific making the diagnosis difficult. In the absence of EEG testing, it is likely to be missed or delayed. When acute brain injury and nonconvulsive status epilepticus occur concurrently, the severity of acute brain injury has traditionally been accepted as determining patient outcome. However, increasing evidence suggests that the two conditions are synergistically detrimental and increase brain injury. Guidelines remain to be established for the intensity and duration of anticonvulsant therapy in these patients. Evidence suggests that, in the absence of extreme and irreversible acute brain injury, early intensive intervention is necessary to improve the otherwise poor outcome of these patients. PMID- 10478707 TI - Assessing the outcomes in patients with nonconvulsive status epilepticus: nonconvulsive status epilepticus is underdiagnosed, potentially overtreated, and confounded by comorbidity. AB - Nonconvulsive status epilepticus (NCSE) is characterized by behavioral or cognitive change from baseline for at least 30 minutes with EEG evidence of seizures. Categorized into complex partial status epilepticus (with lateralized seizures), and generalized nonconvulsive status epilepticus (bilateral diffuse synchronous seizures), there is debate regarding the diagnosis and morbidity of NCSE. Because EEG is needed for diagnosis, only a high index of suspicion leads to a request for the study, whereas EEG is often unavailable after hours or on weekends. Furthermore, the cognitive changes during NCSE are often incorrectly ascribed to a postictal state, intoxication, psychogenic or psychotic states, and mental retardation. Regarding categorization, present classifications address EEG features but fail to distinguish among depths of coma. Deeply comatose patients (with coma etiologies that themselves carry poor prognoses) are mixed with lightly obtunded patients with no morbidity, confusing the prognosis. Thus, a classification that subsumes depth of coma, and possibly etiology, is sorely warranted. Regarding treatment, comatose NCSE patients treated with benzodiazepines may worsen, whereas generalized nonconvulsive status epilepticus patients may suffer iatrogenically from aggressive treatment (hypotension and respiratory depression) necessitating balancing the potential neurologic morbidity of NCSE against the possible morbidity of IV antiepileptic drugs. A high index of suspicion is needed to initiate EEG studies. Better stratification of level of consciousness will be needed to distinguish among morbidity due to underlying conditions, treatment, and the effects of status epilepticus, proper. PMID- 10478708 TI - EEG and clinical associations with mortality in comatose patients in a general intensive care unit. AB - The authors examined EEG findings and clinical factors for their association with outcome in comatose patients in their general intensive care unit. The following individual and combinations of factors were strongly related to mortality, with positive predictive values of >0.80 and odds ratios >2.0: age over 65 years, anoxic/ischemic encephalopathy, EEG suppression, lack of EEG reactivity; anoxia ischemia with partial or complete cranial nerve areflexia, anoxia-ischemia with EEG suppression; anoxia-ischemia and generalized epileptiform activity; anoxia ischemia with partial cranial nerve areflexia and EEG suppression. Conversely, the following factors favored survival rather than death: systemic infection/sepsis, metabolic derangement (excluding anoxic-ischemic insult), trauma; dysrhythmia, focal epileptiform activity, and regional delta and reactivity on EEGs. The findings of this study support the integration of these data into intensive care unit prognostic scoring systems, such as later versions of the Acute Physiology and Chronic Health Evaluation (APACHE). PMID- 10478709 TI - Sources of movement-related cortical potentials derived from foot, finger, and mouth movements. AB - Movement-related cortical potentials (MRCPs) register brain electrical activity before and during movement execution. In an attempt to delineate the components of MRCPs that reflect common sources to various movements and that are movement specific, simple self-paced voluntary foot, finger, and mouth movements were studied. MRCPs were recorded in eight healthy volunteers with 30 electrodes placed on the scalp. Data were analyzed using Brain Electric Source Analysis software, and multiple equivalent dipole models were developed to separate spatial and temporal aspects of brain activity related to the execution of voluntary movements. Independent models were separately developed for the grand average data and for the individual subjects' data for each movement type. MRCPs derived from foot movements were accounted for using a 5-dipole model, finger movements using an 8-dipole model, and mouth movements with a 7-dipole model, yielding the grand average residual variances of 3%, 2%, and 6%, respectively. Based on individual models, intersubject variability of dipole locations was less than 10 mm (+/- SD). Overlaying the mean dipole coordinates onto the stereotaxic atlas provided proof that the sensorimotor cortical areas, supplementary motor area, and also cerebellum and thalamus were active in all three movements. Locations of the dipoles in the contralateral sensorimotor area clearly implied well-known medial to lateral somatotopic organization of foot, finger, and mouth movements. Temporal separation of the activity spread over different brain areas was demonstrated by evolution in the moments of dipole source potentials. The authors' models support the view of simultaneous activation of the primary motor cortex and supplementary motor area at the time of movement execution. Multiple equivalent dipole models developed in this study implied the activity originating in corresponding brain areas as previously detected by positron emission tomography or functional magnetic resonance imaging. However, MRCPs provided additional information regarding the temporal evolution of the brain activity related to the execution of voluntary movements. Thus, the concurrent use of MRCPs and other imaging techniques may provide complementary information not easily obtained by the other imaging techniques themselves. PMID- 10478710 TI - Enhancement of left-right sensorimotor EEG differences during feedback-regulated motor imagery. AB - EEG feedback studies demonstrate that human subjects can learn to regulate electrocortical activity over the sensorimotor cortex. Such self-induced EEG changes could serve as control signals for a Brain Computer Interface. The experimental task of the current study was to imagine either right-hand or left hand movement depending on a visual cue stimulus on a computer monitor. The performance of this imagination task was controlled on-line by means of a feedback bar that represented the current EEG pattern. EEG signals recorded from left and right central recording sites were used for on-line classification. For the estimation of EEG parameters, an adaptive autoregressive model was applied, and a linear discriminant classifier was used to discriminate between EEG patterns associated with left and right motor imagery. Four trained subjects reached 85% to 95% classification accuracy in the course of the experimental sessions. To investigate the impact of continuous feedback presentation, time courses of band power changes were computed for subject-specific frequency bands. The EEG data revealed a significant event-related desynchronization over the contralateral central area in all subjects. Two subjects simultaneously displayed synchronization of EEG activity (event-related synchronization) over the ipsilateral side. During feedback presentation the event-related desynchronization/event-related synchronization patterns showed increased hemispheric asymmetry compared to initial control sessions without feedback. PMID- 10478711 TI - Functional imaging of postmovement beta event-related synchronization. AB - The linear estimation (LE) and spline surface Laplacian (SL) method were applied to single-trial EEG data. EEG was recorded in three subjects during voluntary, self-paced movements of the index finger. The EEG data were bandpass-filtered in the lower beta frequency range and showed short-lasting bursts of oscillations after termination of movement. These oscillations are termed postmovement beta synchronization. The realistic head geometry and the digitized positions of the electrodes were taken into account for accurate modeling of the anatomy. Regularization of the LE method was achieved by the truncated singular value decomposition. The LE and SL distribution of the postmovement beta synchronization showed similar spatial and temporal patterns. A clear increase of the LE source activity was found over the primary motor area. These results indicate that the postmovement beta synchronization is generated over the anterior bank of the central sulcus. PMID- 10478712 TI - Visual evoked oscillatory responses of the human optic tract. AB - Optic tract oscillatory responses directly recorded during posteroventral pallidotomy were investigated to reveal their features with respect to extracranially recorded visual oscillations and to clarify their contributions to scalp-recorded or far-field visual evoked potentials. Oscillatory responses of the optic tract consisting of early and subsequent late oscillations were recorded in all patients. Early oscillations consist of five negative and positive peaks, and late oscillations consist of two to four negative and positive peaks. The frequency of the first peak of early oscillations (103.0+/ 9.2 Hz, n = 14) was significantly lower than that of others (t test, P<0.006), but there were no significant differences among other peaks (t test, P>0.4). This difference was not observed among peaks of late oscillations (t test, P>0.3). As a whole, the frequency of early oscillations (123.9+/-16.7 Hz, n = 70) was significantly higher than that of late oscillations (66.3+/-13.7 Hz, n = 41) (t test, P = 0.0001). Intracerebral recording of the optic responses with the same band filter of scalp-recorded visual evoked potentials (10 Hz to 1 KHz) showed prominent negative (No) and positive (Po) waves, whereas responses obtained over the scalp at Oz and Cz consisted of negative (NI), positive (PI), negative (NII), and positive (PII) waves. Comparisons between No and PI and Po and NII showed overall phase reversal relations between them, but there were significantly different peak latencies between them (t test, P<0.001) except that between Po (116.7+/-11.7 msec, n = 5) and NII of Cz recording (118.4+/-9.4 msec, n = 5) (t test, P<0.3). Our study in conjunction with other studies on visual oscillations revealed that a relatively constant frequency of oscillations is traveling in the subcortical visual system and is probably playing an important role in generating stationary or fixed peaks of the far-field potentials of visual evoked potentials. PMID- 10478713 TI - Direct effects of cyclosporin A on proliferation of hematopoietic stem and progenitor cells. AB - Cyclosporin A (Cy A) has been reported to both stimulate and inhibit bone marrow colony assays in a dose-dependent manner. The observation that anti-gamma-IFN antibodies stimulate hematopoiesis to the same degree as Cy A has led several groups to propose that the stimulatory effects of Cy A are due to inhibition of gamma-IFN production by T cells. In this study we observed that cultures of highly enriched hematopoietic stem/progenitor cells (HSPC), devoid of CD3/5/8+ T cells, also exhibit enhanced cloning efficiency when cultured in the presence of Cy A. Normal bone marrow cells or Thy-1.1lowSca-1+Lin(neg) HSPC were incubated in methylcellulose cultures and stimulated with various combinations of steel factor (SF), interleukin (IL)-3, IL-6, granulocyte colony stimulating factor (G-CSF), and erythropoietin (EPO) in the presence of increasing concentrations of Cy A. HSPC cultures with SF, IL-3, and IL-6 stimulation and low Cy A concentrations had from 24% to 78% higher cloning efficiencies than did parallel cultures without Cy A, and did not fall below control levels until the Cy A concentration was increased to more than 1.25 microg/ml. The addition of EPO and G-CSF abrogated the Cy A stimulation observed with SF, IL-3, and IL-6. These results were reflected in whole bone marrow, but with a higher range of variability. Cultures in which FK-506 replaced Cy A showed no consistent stimulation or inhibition of colony formation. These studies show that Cy A can stimulate hematopoietic stem cell growth independent of mediation by T cells. Consequently, these results argue for a direct positive effect of Cy A on the signal transduction pathways in HSPC. PMID- 10478714 TI - Development of poly(propylene fumarate-co-ethylene glycol) as an injectable carrier for endothelial cells. AB - Poly(propylene fumarate-co-ethylene glycol) [P(PF-co-EG)] hydrogels were examined as in situ polymerizable carriers for endothelial cells. The temperature increase from 37 degrees C during cross-linking was measured. The maximum temperature did not increase above 38.3 degrees C for any copolymer formulation. The temperature profiles also appeared to be independent of the amount or molecular weight of poly(ethylene glycol). These materials were polymerized in situ in a subcutaneous rat model and evaluated for initial biocompatibility. A normal wound-healing response was seen with formation and subsequent maturity of a fibrous capsule. Endothelial cells were embedded in vitro during the cross-linking process and their proliferation was assessed over the first 24 h. There was significant DNA synthesis by the embedded endothelial cells during this time period. These data suggest that P(PF-co-EG) hydrogels could be developed for use as injectable cell carriers. PMID- 10478715 TI - Design engineering of a bioartificial renal tubule cell therapy device. AB - The use of a bioartificial renal tubule device composed of renal proximal tubule cells grown within a hollow fiber cartridge is a first step in engineering a bioartificial kidney to provide more complete replacement therapy of renal function than is available today. In this study, the feasibility of two designs for a tubule device were investigated: one with cells grown on microcarrier beads densely packed within the extracapillary space of a hollow fiber cartridge, and the other with cells grown as a confluent monolayer within the hollow fibers themselves. First, the oxygen requirements of porcine renal proximal tubule cells were determined, both attached to microcarriers and in suspension and compared to that of proximal tubule segments. The basal rate of cell respiration was found to be 2.29 +/- 0.53 nmol O2/10(6) cells/min for our cultured proximal tubule cells in suspension and no significant difference was seen with attached cells. Proximal tubule segments displayed significantly higher respiratory rates. Cells were also found to be responsive in the presence of mitochondrial inhibitors or uncouplers, and their respiratory rates remained constant, despite multiple passaging. The resultant cell oxygen consumption parameter was used in models describing oxygen concentration profiles within the two device configurations. From these models, it was found that cells within our proposed device designs could theoretically be sustained and remain viable, with respect to oxygen limitations. Finally, flow visualization studies were performed to assess fluid flow distribution and determine optimal device configuration and geometry to decrease areas of low or stagnant flow. PMID- 10478716 TI - Analysis of the Th1/Th2 paradigm in transplantation: interferon-gamma deficiency converts Th1-type proislet allograft rejection to a Th2-type xenograft-like response. AB - The rejection mechanisms for fetal proislet allografts and pig proislet xenografts in mice are characterized by different intragraft cytokine mRNA profiles and cellular responses. Allograft rejection is predominantly CD8 T-cell dependent and is associated with a Th1-type cytokine pattern (i.e., IFN-gamma, IL 2 but no IL-4 or IL-5 mRNA). In contrast, xenograft rejection is CD4 T-cell dependent and is accompanied by a strong Th2-type response (i.e., enhanced expression of IL-4 and IL-5 mRNA) and by marked eosinophil accumulation at the graft site. We have now examined and compared the regulatory role of IFN-gamma in both proislet allograft and xenograft rejection processes. The histopathology and intragraft cytokine mRNA profile of BALB/c (H-2d) proislet allografts were examined in IFN-gamma-deficient and wild-type C57BL/6J recipient mice. The survival of pig proislet xenografts was also assessed in IFN-gamma -/- and wild type hosts. Both proislet allografts and xenografts were acutely rejected in IFN gamma -/- and wild-type mice. Unlike the conventional allograft reaction, which lacks eosinophil infiltration, the rejection of proislet allografts in IFN-gamma deficient hosts correlated with intragraft expression of IL-4 and IL-5 mRNA (i.e., a Th2-type response) and eosinophil recruitment. The rejection of proislet allografts and xenografts can therefore occur by IFN-gamma-independent pathways; IFN-gamma, however, regulates the pathology of the allograft reaction but not the xenograft response. The immune destruction of proislet allografts is not prevented by Th2 cytokine gene expression; instead, the latter correlated with the recruitment of unconventional inflammatory cells (eosinophils), which may play an accessory role in effecting graft injury. Significantly, the Th1-to-Th2 like switch resulted in the novel conversion of an allograft rejection reaction into a xenograft-like rejection process. PMID- 10478717 TI - Regulatory T cells maintain peripheral tolerance to islet allografts induced by intrathymic injection of MHC class I allopeptides. AB - Although transplantation remains the treatment of choice for diabetes mellitus, immunological rejection of allografts continues to be a major problem. The search for strategies to prevent graft rejection led us to examine if the fate of developing T cells may be influenced by the presence of allo MHC class I peptides in the thymus because T cell receptor-MHC class I/self-peptide interaction regulates thymocyte development. We studied the effects of intrathymic (IT) injection of a short segment of a synthetic immunogenic MHC class I peptide (peptide 2, residues 67-85) of the hypervariable domain of RT1.A derived from WAG rat (RT1U) on islet graft survival in the WF(RT1U)-to-ACI combination. Adult diabetic male recipients were treated with IT injection of a single WAG-derived MHC class I peptide 7 days before intraportal islet transplantation. Long-term unresponsive islet recipients were examined for the development of alloantigen (Ag)-specific regulatory cells. The results showed that while IT injection of 150 microg peptide 2 on day -7 did not prolong graft survival in naive recipients [median survival time (MST) of 14.0 days vs. 9.6 in controls], IT injection of 300 or 600 microg peptide 2 led to normoglycemia and permanent islet survival (> 200 days) in 4/6 and 3/5 STZ-induced diabetic ACI recipients, respectively. IT injection of 150, 300, or 600 microg peptide 2 combined with 0.5 antilymphocyte serum (ALS) immunosuppression on day -7 led to 100% permanent islet allograft survival (> 200 days) compared to MST of 15.0 +/- 2.3 days in ALS alone-treated controls. Similarly prepared animals rejected third-party Brown Norway (BN) islets in an acute fashion, thus demonstrating donor specificity. Intravenous injection of 300 microg peptide 2 combined with 0.5 ml ALS did not prolong islet allograft survival. The long-term unresponsive islet allograft recipients challenged with second set grafts accepted permanently 100% donor-type cardiac allografts while rejecting third-party (BN) hearts without rejecting the primary Wistar Furth (WF) islets. In analyzing the underlying mechanisms of acquired systemic tolerance, we found no suppressor/regulatory cells in adoptive transfer studies in tolerant animals at 30 days after IT injection of allopeptides. In contrast, adoptive transfer of 5 x 10(7) unseparated spleen cells from tolerant animals at 60 and 100 days after islet transplantation into lightly irradiated [200 rad total body irradiation (TBI)] ACI recipients led to donor-specific permanent islet graft survival in 2/3 and 4/5 secondary recipients, respectively, compared to an MST of 13.8 days in lightly irradiated ACI given unmodified syngeneic spleen cells. In addition, adoptive transfer of 2 x 10(7) purified T cells obtained from long-term functioning islet recipients led to permanent donor specific islet survival in secondary recipients. The finding that IT injection of a short segment of a synthetic immunodominant MHC class I peptide derived from WAG that shares the RT1.A(U) domain with the graft donor is capable of inducing acquired systemic tolerance to WF islets suggests that linked recognition or epitope suppression may be involved in the induction of unresponsiveness. Generation of peripheral Ag-specific regulatory cells that suppress Ag-specific alloreactive T cells is, in part, responsible for the maintenance of tolerance in this model. PMID- 10478718 TI - Forskolin-stimulated adenylylcyclase activity: a marker to assess islet cell viability following cold storage in different solutions and to predict islet cell function following transplantation. AB - For clinical islet cell transplantation, short-term storage of islet cells is likely to be necessary, and it is imperative that the islet cells be kept as viable as possible during the period. However, there are little data on which preservative solutions are most suitable for the storage of islet cells after isolations or before transplantation. To estimate islet cell viability and transplantation success rate in the present study, adenylylcyclase activity was measured with a rapid new fluorometric assay in rat islet cells prior to transplantation, because cAMP plays an essential role in determining islet beta cell viability and responsiveness to various hormonal stimuli. Adenylylcyclase activity was measured in islet cells stored for different periods of time 0, 3, 16, 24, 48, 96 h) and in different preservative solutions. Approximately 1,000 islet cells from each preservation group using University of Wisconsin (UW) solution were transplanted to streptozotocin-induced diabetes mellitus (DM) rats. Transplant success was evaluated by measuring blood glucose levels. Preoperative adenylylcyclase activity was compared with posttransplant islet cell function. The adenylylcyclase activity of UW solution was significantly higher than that of Euro-Collins solution and lactate-Ringer's solution through the different preservation time periods. Preoperative adenylylcyclase activity correlated well with posttransplant islet cell function in a rat model of DM. We conclude that adenylylcyclase activity can be used as a marker to assess islet cell viability as well as differences in preservation media and may predict islet cell transplant success. PMID- 10478719 TI - Novel method for isolation of adult porcine pancreatic islets with two-stage digestion procedure. AB - It is particularly difficult to isolate porcine islets (PI). Experience suggests that the success rate of porcine islet isolation (PII) is probably considerably influenced by the distension and digestion of the pancreas. In this study, we divided the digestion procedure into two stages and developed a new enzyme solution to improve both the distension and digestion procedures. As a result, we established a novel and stable method of large-scale adult porcine islet isolation (APII). The harvested pancreata of 2-year-old pigs weighing over 200 kg (n = 18) were distended by introducing our new enzyme solution gently and slowly through the pancreatic ducts. Two-stage digestion (cold, then warm) was then performed by first placing the distended pancreata on ice for 2 h to cause diffusion of the enzyme solution around the islets, and then by incubating the pancreata in a water bath at 37 degrees C for 45 min without shaking. The islets were purified by a COBE 2991 cell processor on dextran T70 discontinuous density gradients. Histological study was performed on porcine pancreata sampled after 0, 15, 30, and 45 min of the second stage, and stained with H&E stain. Next, islet equivalent was calculated. Static incubation study was performed by stimulating the islets with 3.3 and 16.7 mM glucose in Krebs' Ringer bicarbonate buffer (KRBB) solution at 37 degrees C for 1 h, and finally the insulin released was measured. The dilated acinar cells septa around the islets were observed at time 0. Destruction of the acinar cells around the islets by warm digestion was recognized at 15 and 30 min, and destroyed and separated acinar cells present around the islets at 45 min. During the entire course of the warm digestion, the islets remained intact. The number of isolated islets was 291,667 +/- 240,452 IEQ/pancreas (n = 14) and 3,294 +/- 2199 IEQ/g of pancreatic tissue. The purity of recovered porcine islets was over 90%. The concentration of the insulin secreted by 10,000 IEQ islets selected at random was 83.9 +/- 13.4 microU/dish/h in response to 3.3 mM glucose and 104.1 +/- 12.9 microU/dish/h in response to 16.7 mM glucose (n = 20). A success rate of approximately 80% was attained with APII. We demonstrated that this increase in the success rate was due to the improved distension and digestion provided by this method. This two-stage APII method with its new enzyme solution may facilitate the future use of porcine islets in clinical xenotransplantation trials. PMID- 10478720 TI - Bioartificial endocrine pancreas (Bio-AEP) for treatment of diabetes: effect of implantation of Bio-AEP on the pancreas. AB - Recently, we described a diffusion chamber for a bioartificial endocrine pancreas (Bio-AEP). Pancreatic islet cells in the Bio-AEP device were isolated from the immune system of the host by an artificial barrier, while nutrients, electrolytes, oxygen, and bioactive secretory products were exchanged across this barrier. This experiment was designed to evaluate whether the diffusion chamber could be useful as a Bio-AEP in the treatment of diabetes. Six streptozotocin (STZ)-induced diabetic rats each received a diffusion chamber containing 8 x 10(6) MIN6 cells as a xenograft Bio-AEP. In the STZ diabetic rats with Bio-AEPs, a return to normoglycemia was observed up to 30 weeks after implantation, without the use of any immunosuppressant. A gradual increase in the body weight of the rats was also observed. In three STZ diabetic rats, diffusion chambers without MIN6 cells were implanted as a sham operation. The fasting blood glucose levels in these three rats remained higher than 600 mg/dl, after implantation, and they lost weight. Thirty-five weeks after implantation, the pancreata were removed from the rats that underwent xenoimplantation, those that had the sham operation, and the normal control rats. In the sham-operated animals, the exocrine tissues of the pancreata were vacuolated and pancreatic B cells were not seen in the islets. In contrast, in the pancreata from the xenoimplantation, the exocrine tissues were normal, and a few pancreatic B cells were seen in the islets. These results indicated that xenoimplantation using the Bio-AEP might retard the progress of diabetes. PMID- 10478721 TI - Effectiveness of acidic oxidative potential water in preventing bacterial infection in islet transplantation. AB - At a number of points in the current procedures of islet isolation and islet culture after the harvesting of donor pancreata, microorganisms could potentially infect the islet preparation. Furthermore, the use of islets from multiple donors can compound the risks of contamination of individual recipients. Acidic oxidative potential water (also termed electrolyzed strong acid solution, function water, or acqua oxidation water), which was developed in Japan, is a strong acid formed on the anode in the electrolysis of water containing a small amount of sodium chloride. It has these physical properties: pH, from 2.3 to 2.7; oxidative-reduction potential, from 1,000 to 1,100 mV; dissolved chlorine, from 30 to 40 ppm; and dissolved oxygen, from 10 to 30 ppm. Because of these properties, acidic oxidative potential water has strong bactericidal effects on all bacteria including methicillin-resistant Staphylococcus aureus (MRSA), viruses including HIV, HBV, HCV, CMV, and fungi as a result of the action of the active oxygen and active chlorine that it contains. We conducted this study to evaluate the effect of acidic oxidative potential water irrigation on bacterial contamination on the harvesting of porcine pancreata from slaughterhouses for islet xenotransplantation by counting the number of pancreatic surface bacteria using the Dip-slide method, and on the results of islet culture; and to evaluate the direct effect on isolated islets when it is used to prevent bacterial contamination by the static incubation test and by morphological examination. Direct irrigation of the pancreas by acidic oxidative potential water was found to be very effective in preventing bacterial contamination, but direct irrigation of isolated islets slightly decreased their viability and function. PMID- 10478722 TI - Beneficial effects of immunoisolated fetal and neonatal pig liver fragments on acute liver failure in a large animal. AB - Xenogeneic cell (fragment) transplantation may be used as an interim therapy until the organ allotransplanation. Immunologic rejection, however, constitutes the major hurdle. To overcome this problem, "xeno" fetal and neonatal liver fragments (FLF, NLF) were encapsulated into separate micropore devices that protect them from immunological attack by the recipient. The FLF or NLF were then transplanted into beagles with hepatic failure to observe their biological effects. In Experiment 1 (n = 5) beagles were injected IV with D-galactosamine (D gal, 1.0 g/kg) on day 0 and then received FLF grafts (0, 0.3, 0.8, 1.0, 2.0 g/kg). In Experiment 2 (n = 6) beagles received NLF grafts (1.8 g/kg) and on the following day were injected with D-gal (1.0 g/kg). In Experiment 1 only the high dose of xeno-FLF (2.0 g/kg) decreased the elevated ALT (GPT) and T. Bil. levels. Histologic examination showed that some of the hepatocytes of the host liver survived only in the high-dose graft. In Experiment 2, at 36 and 48 h after D-gal injection, the transplanted group had a significantly lower AST (GOT) level than the control. The grafted NLF survived for 14 days, according to histologic examinations. Thus, encapsulated FLF and NLF xenotransplantation can prevent liver dysfunction in a large animal hepatic failure model. PMID- 10478723 TI - Protective effect of nafamostat mesilate on injury of porcine hepatocytes by human plasma. AB - Nafamostat mesilate (NM), a protease inhibitor, possesses a cytoprotective effect and inhibits the activation of complement. The present study investigated whether NM has any protective effect against injury of porcine hepatocytes by human plasma in a bioartificial liver support system. Porcine hepatocytes were harvested and seeded at a density of 2 x 10(5) cells on a 35-mm collagen-coated plate in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal calf serum. Twenty-four hours later, the medium was replaced with human plasma with three concentrations of NM between 3.8 x 10(-5) and 3.8 x 10(-4) M and then cultured for 6 h. The viability of porcine hepatocytes, lactate dehydrogenase (LDH) levels, lidocaine clearance, porcine albumin production, and changes in complement (C3) levels were measured. The viability of porcine hepatocytes in human plasma decreased significantly to 37.7 +/- 11.4% of that in DMEM. NM improved the viability of the hepatocytes, lowered the levels of LDH, and increased lidocaine clearance and albumin production in a concentration-dependent manner. The concentrations of C3, the marker of xenogeneic reactions, did not change significantly, indicating that no hyperacute xenogeneic reaction occurred in our series. Together, our results suggested that NM exerts favorable effects on porcine hepatocytes in human plasma through direct effect such as prevention of protease activity in the plasma membrane of porcine hepatocytes rather than inhibition of complement-dependent immunoreactions. PMID- 10478725 TI - Long-term functional assessment of encapsulated cells transfected with Tet-On system. AB - In our previous study, xenogeneic mouse neuroblastoma cells bearing the POMC gene, the precursor of ACTH and beta-endorphin, were implanted within polymer capsules into the CSF space of rats. Although ACTH and beta-endorphin were secreted, we were not able to control the amounts or times of hormone release. A promoter that is inducible by administration of tetracycline derivatives (Tet) was linked to the POMC gene to control its gene expression (Neuro2A-Tet-On-POMC; NTP). The results showed that POMC gene expression in the implanted encapsulated NTP cells could be regulated in a dose-dependent manner by Tet administration to the hosts. However, no analysis of gene control with the Tet-On system over a long period has been performed. In this study, encapsulated NTP cells were treated in vitro with doxycycline (Dox) (1.0, 10, 100, 1000 ng/ml) continuously for a month. On day 4, the amount of ACTH secretion was dependent on the Dox dose. But in the course of the experiment, the difference of ACTH secretion among those treated with Dox 10, 100, and 1000 ng/ml was eliminated. On the other hand, NTP cells, which were treated with Dox (1000 ng/ml) just on days 7, 14, 21, and 28, secreted almost the same amount of ACTH in 24 h. From these results, for clinical use, an NTP cell line that secretes enough opiate to reduce pain sensitivity without Dox should be established, and Dox could then be administered if necessary. PMID- 10478724 TI - Evaluation of reaction of primate brain to grafted PC12 cells. AB - Intrastriatal implantation of polymer-encapsulated PC12 cells, which constitute a dopaminergic cell line derived from rat pheochromocytoma, has proved useful for ameliorating parkinsonian symptoms in several kinds of animals. In considering the clinical application of this technique, we should make sure that PC12 cells are rejected completely by the host immune system in case the capsule breaks. In the present study, unencapsulated PC12 cells were injected into the brain of Japanese monkeys (Macaca fuscata). Histological [hematoxylin-eosin (H&E), Nissl] and immunocytochemical [tyrosine hydroxylase (TH), and glial fibrillary acidic protein (GFAP)] analyses were performed 1, 2, 4, and 8 weeks after transplantation. Also, encapsulated PC12 cells were transplanted into the brain of another group of Japanese monkeys to investigate the host reaction to the capsule and to confirm that the encapsulated PC12 cells continue to survive in the host brain. H&E and GFAP staining were performed 2, 4, and 8 weeks after transplantation. L-DOPA and dopamine release from the explanted capsules was measured by high performance liquid chromatography. Magnetic resonance imaging was performed in both unencapsulated and encapsulated PC12 cell grafted groups. Although the xenografted unencapsulated cells formed a small cluster at 1 and 2 weeks after implantation, very few and no viable PC12 cells remained at 4 and 8 weeks, respectively. The reaction of the host towards the xenograft gradually decreased. Encapsulated PC12 cells retrieved from the host brain were found to release L-DOPA and dopamine continuously even 8 weeks after implantation. The host reaction to the PC12-loaded capsule was much weaker than that to the unencapsulated PC12 cells, and decreased with time. These results indicate that encapsulated PC12 cell transplantation is an effective and safe strategy for the treatment of Parkinson's disease. PMID- 10478726 TI - Differentiation and angiogenesis of central nervous system stem cells implanted with mesenchyme into ischemic rat brain. AB - After cerebral infarction, necrosis in neural tissues is not usually repaired or reconstructed by the injured brain. We therefore examined the effects on postinfarction repair of implanting central nervous system (CNS) stem cells together with mesenchyme, because CNS stem cells can be expected to adapt and survive in the adult brain. Cerebral infarction was induced by the Koizumi-Longa method, using the adult male spontaneous hypertensive rat model. Reperfusion was performed an hour after middle cerebral artery occlusion. The rat mesencephalic neural plate at the early somite stage (embryonic day 10.5) together with the adjacent ventral mesenchymal tissues was dissected out under the microscope and immediately implanted into the ischemic rat striatum. One month later, the cognitive function was evaluated by the Morris water maze method. Histologic and immunohistochemical examinations of the graft were made with hematoxylin-eosin (H&E), neurofilament-200, and tyrosine hydroxylase (TH) stains. In the water maze study, mean latency times required to reach an escape platform in the implanted animals with surviving grafts were found to be shorter than in those without grafts, but longer than in normal animals. In the spatial probe trial, the number of animals seen to cross the area in the pool where the platform had been located was greater in the implanted rats with surviving grafts than in other groups. Multiple vascularization in the grafted area was observed histologically in H&E stained tissues, and neurofilament-200-positive cells were recognized in the graft. TH staining revealed within the graft many immunoreactive neuron-like cell bodies with long dendrites. It was suggested that grafted CNS stem cells with mesenchyme may survive and differentiate into mature CNS tissue within the adult ischemic rat brain, constructing vessels in and around the grafts, and may therefore have the potential to be effective in the recovery of the cognitive function of the rat model. PMID- 10478727 TI - Chromosomal abnormalities in a series of children with autistic disorder. AB - In a series of 127 children diagnosed with autistic disorder the karyotypes of 8, on whom data were available, showed the following chromosomal abnormalities: breakage, a 47 XY pattern, trisomy 13, inversion-duplication of chromosome 15, 47 XY, +der (15) (pter q15: p11 pter), 47 XXY and 46 XY, inv (2) (p11:q13pat, 3q+). Compared to those who were not karyotyped or had normal karyotypes, the children with abnormalities, although cognitively more delayed, were not rated as more severely autistic. Facial dysmorphias and minor physical anomalies tended to be more frequent in the chromosomally deviant subgroup. No differences in demographic characteristics or parental ages were evident. Results are consistent with the view of variability of expression of marker chromosome deviations and a greater severity of retardation and symptoms of autism in those affected. The relevance of the findings to a multimodal genetic etiology of autistic disorder is discussed. PMID- 10478728 TI - A comparison of social skills in adults with autistic disorder, pervasive developmental disorder not otherwise specified, and mental retardation. AB - We examined the social skills of adults with autism, PDDNOS, and mental retardation. All participants were diagnosed with profound mental retardation. Participants in the autism and PDDNOS groups had been previously diagnosed using the Childhood Autism Rating Scale (CARS) and record review. These diagnoses were confirmed by readministering the CARS by one author and an independent rater. Social skills were assessed by using the Vineland Adaptive Behavior Scales and the Matson Evaluation of Social Skills in the Severely Retarded. Significant differences between the autism and mental retardation groups were found on both measures. The PDDNOS group demonstrated better positive nonverbal social skills than the autism group but not the mental retardation group. Special treatment needs of institutionalized adults with autism appear warranted along with a need to clarify further the differences between PDDNOS and mental retardation. PMID- 10478729 TI - Parental perspectives on inclusion: effects of autism and Down syndrome. AB - This study examined the effects of the child's diagnosis (autism vs. Down syndrome), age, and current educational placement on parental perceptions toward inclusion for their child with disabilities. Parents of children with autism and with Down syndrome completed surveys regarding their opinions on their child's current educational placement, their desire for changing the current placement, and their views on inclusive education. Results indicated that diagnosis, age, and current placement influenced parental opinion on the ideal educational placement for their child. Parents of children with Down syndrome were significantly more likely to endorse inclusion (full-time placement in general education) as the ideal educational program for their child whereas parents of children with autism were more likely to endorse mainstreaming (consistent part time placement with general education students). Parents of younger children and parents whose children were already placed in general education programs were more positive towards inclusion than parents of older children or students currently in special education. Findings are discussed in terms of child characteristics and prevailing educational practices. PMID- 10478730 TI - Social and nonsocial factors in the Childhood Autism Rating Scale. AB - The Childhood Autism Rating Scale (CARS) was factor analyzed to determine if distinct and independent "subgroups" of symptoms could be derived, which would be consistent with the current multidimensional theory and nosology for autism. To address this issue, the CARS was factor analyzed for a sample of 90 children with diagnoses of either autism or PDDNOS, based on DSM-III-R diagnostic criteria. Five factors emerged: Social Communication, Emotional Reactivity, Social Orienting, Cognitive and Behavioral Consistency, and Odd Sensory Exploration. Factor-based scales were created. These factor-based scales were demonstrated to distinguish subjects with autism from subjects with PDDNOS and nonautistic subjects. Factor-based scores were examined to determine the degree to which they were associated with individual differences (such as age, IQ, gender, history of regression, and history of abnormal EEGs) among children with pervasive developmental disorders (PDDs). The application of these distinct and independent factors may have important clinical and research implications. The generation of factor-based scales may provide information on the nature of the individual differences that are thought to be present among children with autism. Additionally, the use of factor-based scale scores may increase the sensitivity of the CARS for identifying younger and/or higher functioning individuals within the PDD spectrum. PMID- 10478731 TI - Creativity and imagination in autism and Asperger syndrome. AB - Three studies are reported that address the often described impoverished creativity in autism. Using the Torrance Creativity Tests, Experiment 1 found that children with autism and Asperger syndrome (AS) showed impairments. Experiment 2 tested two explanations of these results: the executive dysfunction and the imagination deficit hypotheses. Results supported both hypotheses. Children with autism and AS could generate possible novel changes to an object, though they generated fewer of these relative to controls. Furthermore, these were all reality-based, rather than imaginative. Experiment 3 extended this using a test of imaginative fluency. Children with autism and AS generated fewer suggestions involving attribution of animacy to foam shapes, compared to controls, instead generating reality-based suggestions of what the shapes could be. Although this is evidence of executive dysfunction, it does not directly account for why imaginative creativity is more difficult than reality-based creativity. PMID- 10478732 TI - Brief report: autism and Asperger syndrome in seven-year-old children: a total population study. PMID- 10478733 TI - Brief report: gaze behavior and theory of mind abilities in individuals with autism, down syndrome, and mental retardation of unknown etiology. PMID- 10478734 TI - Parental advocacy. PMID- 10478735 TI - Alfred Kinsey: Asperger disorder. PMID- 10478736 TI - Are measles infections or measles immunizations linked to autism? PMID- 10478737 TI - Pain palliation with rhenium-186 HEDP in breast cancer patients with disseminated bone metastases. AB - PURPOSE: For patients with metastatic prostate cancer, first results have shown that rhenium-186 (Re-186) hydroxyethylidene diphosphonate (HEDP) is efficient in pain palliation of disseminated bone metastases. The aim of this study was to determine whether significant pain reduction can also be achieved in breast cancer patients with Re-186 HEDP. METHODS: Thirty patients with breast cancer who had multifocal painful bone metastases received a total of 38 intravenous Re-186 HEDP injections. Pain relief was assessed through daily documentation of the visual analog scale and analgesic consumption. A significant response to treatment was determined if the visual analog scale or analgesic consumption decreased significantly for at least 2 weeks. Blood counts were controlled at baseline and at weeks 4 and 8. RESULTS: A response to pain therapy was observed in 60% (18 of 30) of the patients. A reversible thrombocytopenia and leukopenia of grade 2 (according to World Health Organization criteria) was found in 4 and 2 patients, respectively. CONCLUSION: Patients with disseminated osseous metastases resulting from breast cancer can benefit from therapy with Re-186 HEDP. PMID- 10478738 TI - Comparison of the dynamics of bile emptying by quantitative hepatobiliary scintigraphy before and after cholecystectomy in patients with uncomplicated gallstone disease. AB - PURPOSE: Quantitative hepatobiliary scintigraphy, a noninvasive method frequently used to diagnose several biliary tract disorders, shows abnormalities in bile secretion and outflow. It is well known that there are wide variations in the normal pattern of bile emptying, but the effect of cholecystectomy on the bile flow has not yet been investigated. The goal of the current study was to examine the dynamics and normal variations of bile flow by quantitative hepatobiliary scintigraphy before and after cholecystectomy in a group of patients with uncomplicated gallstone disease. METHODS: Twenty patients were evaluated before and after cholecystectomy through cholecystokinin octapeptide-augmented quantitative hepatobiliary scintigraphy, and quantitative parameters of bile emptying (Tmax: time to peak activity, T1/2: half-emptying time before and after cholecystokinin octapeptide and duodenum appearance time) were determined and then compared. RESULTS: Before operation, the bile outflow displayed wide variations, with a moderately delayed common bile duct emptying time in some patients. After cholecystectomy, the T1/2 of the common bile duct decreased significantly when compared with the preoperative status, with only minor patient to-patient variation, indicating uniformly faster bile emptying (common bile duct T1/2 before and after operation: 30.5 +/- 14.8 and 18.8 +/- 2.6 min, respectively). Cholecystokinin octapeptide administration caused rapid bile outflow from the common bile duct, with a significant decrease in the T1/2 parameters before and after cholecystectomy. CONCLUSIONS: In patients with their gallbladders in situ, the bile emptying rate showed wide variations and may be moderately slow without distal common bile duct obstruction. After cholecystectomy, the rate of bile emptying accelerated and showed only minor variations, thereby increasing the sensitivity of quantitative hepatobiliary scintigraphy for showing partial biliary obstruction. PMID- 10478739 TI - The use of hepatocyte extraction fraction to evaluate neonatal cholestasis. AB - PURPOSE: Hepatobiliary scintigraphy is used routinely to evaluate infants with neonatal cholestasis. Hepatobiliary scintigraphy determines biliary patency by detecting radioactivity in the bowel on imaging, in duodenal and gastric aspirates, or all of these. During hepatobiliary scintigraphy, the hepatocyte extraction fraction (HEF) is calculated by deconvolution analysis. Normal values of HEF are more than 90%. It is believed that HEF may predict hepatic dysfunction, because, during hepatobiliary scintigraphy, the radiopharmaceutical used in this test is extracted by the hepatocytes from the blood stream. Therefore, a low value of HEF is seen with more severe hepatocellular disease. The goal of this study was to determine whether HEF has any correlation with synthetic liver function, whether HEF can differentiate obstructive from nonobstructive lesions that cause neonatal cholestasis, and whether HEF can predict the outcome of the different causes of neonatal cholestasis. METHODS: A retrospective analysis of 68 hepatobiliary scintigraphy results was done in patients with neonatal cholestasis for a period covering 6 years. RESULTS: The HEF was available in 67 of these 68 patients, with a median value of 25% (range, 3.3% to 100%). The results of synthetic liver function tests (i.e., albumin and prothrombin time) were normal in all infants with neonatal cholestasis. No significant correlation was detected between HEF and the serum levels of total and direct bilirubin, albumin, alkaline phosphatase, and prothrombin time by exploratory data analysis (R = 0.08; small, P > 0.2). The HEF values in different causes of neonatal cholestasis were compared: extrahepatic biliary atresia, neonatal hepatitis, and a miscellaneous category consisting of alpha1-antitrypsin deficiency, ischemic hepatitis, paucity of bile ducts, and others. The outcomes of these diseases were assessed as resolution, continuing disease, transplantation, or death, but no predictive correlation was found with HEF. CONCLUSIONS: A single determination of HEF is of no value in assessing synthetic liver function (as assessed by albumin and prothrombin time), specific diagnoses, and outcomes in patients with neonatal cholestasis. Therefore, a low isolated value of HEF should not be considered suggestive of poor prognosis and outcome in these patients. PMID- 10478740 TI - Comparison between cortical distribution of I-123 iomazenil and Tc-99m HMPAO in patients with Alzheimer's disease using SPECT. AB - PURPOSE: To compare brain perfusion and synaptic density in Alzheimer's disease assessed using I-123 iomazenil SPECT with brain perfusion assessed using Tc-99m HMPAO SPECT. MATERIALS AND METHODS: Early and delayed I-123 iomazenil SPECT images acquired 20 and 180 minutes after injection were compared with Tc-99m HMPAO SPECT studies acquired 15 to 20 minutes after injection in five patients with Alzheimer's disease. RESULTS: Visual analysis of I-123 iomazenil images showed more severe (n = 4) and extensive (n = 3) defects than did Tc-99m HMPAO. Semiquantitative analysis was performed by normalizing the uptake of Tc-99m HMPAO and I-123 iomazenil in individual brain regions in the cerebellum and expressing these values as a ratio of the occipital regions. The analysis of brain regional ratios in Tc-99m HMPAO studies showed a low but significant correlation with ratios of delayed (r = 0.325, P < 0.05) images in the I-123 iomazenil studies. Furthermore, when compared with Tc-99m HMPAO, early (P < 0.01) and delayed mean ratios (P < 0.05) were significantly less in the frontal regions; early mean ratios were significantly less in the temporal regions (P < 0.05), and delayed (P < 0.05) mean ratios were significantly less in the parietal regions. CONCLUSIONS: Tc-99m HMPAO images were better correlated with I-123 iomazenil images, indicating cortical synaptic density (delayed images). I-123 iomazenil SPECT in patients with Alzheimer's disease provided more sensitive information than Tc-99m HMPAO, allowing evaluation of brain perfusion and synaptic density. PMID- 10478741 TI - Reversible cerebral hypoperfusion observed with Tc-99m HMPAO SPECT in reversible dementia caused by hypothyroidism. AB - A 69-year-old man had hypothyroid dementia as a result of I-131 therapy and an overdose of methimazole. Tc-99m HMPAO SPECT revealed diffuse cerebral hypoperfusion. The findings of brain SPECT normalized with the disappearance of symptoms and a return to the euthyroid state. There was a 25% or 26% reduction of the mean cerebral blood flow during dementia. This may be the first report in which SPECT brain imaging revealed reversible hypoperfusion associated with reversible hypothyroid dementia. PMID- 10478742 TI - The value of Ga-67 renal SPECT in diagnosing and monitoring complete and incomplete treatment in children with acute pyelonephritis. AB - PURPOSE: This study evaluated the utility of Ga-67 renal SPECT for diagnosing acute pyelonephritis (APN) in children and monitoring them. METHODS: Seventy-one children (ages 1 week to 12 years) who were thought clinically to have APN were included in the study. The disease was considered present if the patients had all of the following: fever (38.5 degrees C), pyuria (leukocyte counts/per high-power field > or = 10), and a positive result of a urinary culture or blood culture. Tc 99m DMSA, Ga-67 renal SPECT, and voiding cystourethrography were performed, with informed consent from the patients' parents, within 3 days after hospitalization. Three months after treatment, Tc-99m DMSA and Ga-67 renal SPECT were repeated in those patients who had abnormal results of the initial Ga-67 renal SPECT. RESULTS: In the diagnostic study, Ga-67 renal SPECT was superior to DMSA renal SPECT in detecting lesions (97% vs. 79%). Three children had false-negative results with Ga-67 renal SPECT. Seventeen kidneys were negative with Tc-99m DMSA but positive with Ga-67 renal SPECT. No patients had any Ga-67 uptake on post therapy imaging. However, 32 of 107 kidneys (30%) had permanent renal scars. In these 107 kidneys, 78 (73%) were associated with high-grade vesicoureteral reflux (VUR; VUR grade > or = 3) and 29 (27%) with low-grade or no VUR. CONCLUSIONS: High-grade VUR tends to be associated more with APN than has been reported by others, probably because of an underestimation of APN by ultrasonography or DMSA. Ga-67 renal SPECT is sensitive and useful not only in diagnosis but also for monitoring and follow-up of children with clinical suspicion of APN, especially in those with equivocal results after DMSA renal SPECT studies. PMID- 10478743 TI - Time-optimized carbon-14 breath test for Helicobacter pylori contamination of the stomach. AB - PURPOSE: Evidence that Helicobacter pylori plays an important role in peptic ulcer disease has generated intense interest in the development of various screening strategies. H. pylori can be detected high sensitivity and specificity by the noninvasive carbon-14 (C-14) breath test. METHODS: The authors assessed the cost-effectiveness of screening patients with dyspepsia before endoscopy. Thirty-four patients with dyspepsia were evaluated by the C-14 breath test, and then the authors reevaluated the criteria for the diagnosis of H. pylori. The discrimination obtained by using values of cumulative radioactivity or specific activity of breath samples collected between 10 and 60 minutes were studied. RESULTS: The main feature of H. pylori colonization in the stomach was a rapid increase of carbon dioxide in expired breath within 10 minutes after oral administration of C-14. Performance of the test was equally excellent for 10 minute values and 60-minute cumulative excretion. CONCLUSION: The C-14 breath test is a reliable method for diagnosing H. pylori colonization in the stomach, and the accuracy is not compromised by reducing the sample time to maintain better cost-effectiveness and patient acceptance. PMID- 10478744 TI - Disappointing results with Tc-99m tetrofosmin for detecting medullary thyroid carcinoma metastases comparison with Tc-99m VDMSA and TI-201. AB - PURPOSE: This prospective study assessed the detectability of metastatic lesions by Tc-99m tetrofosmin in medullary thyroid carcinoma and to compare the results with Tc-99m penta dimercapto succinic acid (VDMSA) and TI-201. METHODS: A prospective study was performed on 24 patients (10 men, 14 women; ages 23 to 76 years) with medullary thyroid carcinoma after total thyroidectomy. Five cases were sporadic and 19 were familial. After the injection of 740 MBq (20 mCi) tetrofosmin, 740 MBq (20 mCi) VDMSA and 74 MBq (20 mCi) TI-201, whole-body scans and 5-minute static images of the head, neck, chest, abdomen and pelvis were obtained. All scintigraphic studies were compared with calcitonin levels, radiologic findings, histopathologic results, and clinical follow-up. RESULTS: Thirty-four metastatic sites were detected in 12 patients on the basis clinical, radiologic, and histopathologic findings. Patients were divided into three groups according to the calcitonin levels and scintigraphic findings. Group 1 consisted of patients with elevated calcitonin levels and positive scintigraphic findings. Among 34 metastatic sites, 30 could be detected with VDMSA. Only 21 and 20 metastatic sites could be visualized with TI-201 and tetrofosmin, respectively. All 30 lesions showed intense VDMSA uptake but only faint or no uptake with TI 201 and tetrofosmin. Patients in group 2 were accepted to have micrometastases. In this group, calcitonin was minimally elevated, and the results of all three scintigraphs were negative. Group 3 included patients with true-negative results. All patients had normal calcitonin levels and negative results of scintigraphic studies. CONCLUSIONS: Tetrofosmin has no role in the detection of medullary thyroid carcinoma metastases. These results show that VDMSA is clearly superior to TI-201 and tetrofosmin in the follow-up of patients with medullary thyroid carcinoma. PMID- 10478745 TI - Unusual appearance of thromboembolism on perfusion lung imaging. AB - A 46-year-old woman with advanced ovarian carcinoma had progressive dyspnea and was evaluated with ventilation and perfusion lung imaging. A characteristic pattern of multiple branching perfusion defects of a segmental nature on the perfusion scan suggested tumor microembolism and lymphangitic carcinomatosis. However, in this case, this pattern was associated with pulmonary thromboembolism and was documented by the post mortem examination. Pulmonary thromboembolism should be included among the differential diagnoses in a patient with clinical symptoms and a perfusion scan that reveals multiple branching perfusion defects. PMID- 10478747 TI - Uptake of Tc-99m MDP in shoulder muscle caused by the use of elbow crutches. PMID- 10478746 TI - Differential uptake of TI-201 by small-cell lung cancer in a patient with pneumoconiosis-related pulmonary nodules. AB - A 68-year-old man with pneumoconiosis was thought to have small-cell lung cancer based on the results of a biopsy of a bone tumor. Three pulmonary nodules were observed on a chest radiograph. Compared with a chest radiograph taken 4 months earlier, one of the nodules had grown. It was difficult to differentiate this nodule from pneumoconiosis-related benign pulmonary nodules from the appearance on the chest radiograph and CT. Ga-67 scintigraphy and TI-201 lung SPECT were performed to characterize these nodules. TI-201 SPECT showed differential high uptake in the enlarged nodule, whereas Ga-67 scintigraphy showed equally intense uptake in all these nodules. Transbronchial biopsy of the nodule that showed high TI-201 uptake revealed cancer cell nests against a background of interstitial fibrosis. The pathologic diagnosis was small-cell lung cancer that had developed in lung scar tissue. This case suggests the utility of TI-201 in scintigraphic assessments of pneumoconiosis-related pulmonary nodules when lung cancer is suspected. PMID- 10478748 TI - F-18 fluorodeoxyglucose positron emission tomographic imaging in desmoplastic small round cell tumor of the abdomen. PMID- 10478749 TI - Scintigraphy in severe tenosynovitis of the tibialis posterior tendon. PMID- 10478750 TI - Brain blood flow SPECT: posterior flow deficits in young patients with depression. PMID- 10478751 TI - The plantar aponeurosis exposed. PMID- 10478752 TI - Chilaiditi's syndrome on Tc-99m hexamethylpropylene amineoxime imaging. PMID- 10478753 TI - Prominent myocardial uptake on perfusion lung imaging. PMID- 10478754 TI - False-positive positron emission tomographic scan for recurrent breast cancer resulting from a bee sting. PMID- 10478755 TI - Separation of I-131-positive juxtagastric metastatic thyroid carcinoma from the stomach by simultaneous dual-isotope imaging with I-131 and Tc-99m pertechnetate. PMID- 10478756 TI - Unilateral absence of pulmonary perfusion in Swyer-James syndrome. PMID- 10478757 TI - Demonstration of pleural empyema on Ga-67 planar and SPECT imaging. PMID- 10478758 TI - The importance of stress in varicocele study. PMID- 10478759 TI - Diffuse radiocolloid uptake in the bone marrow on thoracolumbar vertebrae extending to the skull, upper sternum, clavicles, long bones, and the lung secondary to lymphoma infiltration of the bone marrow. PMID- 10478760 TI - Increased F-18 fluorodeoxyglucose thyroidal uptake in Graves' disease. PMID- 10478761 TI - Abnormal lymphatic flow demonstrated by lymphoscintigraphy in chylothorax correlation with lymphography. PMID- 10478762 TI - Marked uptake of TI-201 in intracranial extracerebral cavernous hemangioma. PMID- 10478763 TI - Incidental detection of bilateral kidney calyces in Tc-99m phytate liver and spleen scan. PMID- 10478764 TI - Warthin's tumor and I-131 body scan. PMID- 10478765 TI - False-positive Tc-99m sestamibi examination for parathyroid adenoma in a case of asymmetrical salivary gland enlargement. PMID- 10478766 TI - Current readings in nuclear medicine. PMID- 10478767 TI - Vertebral artery volume flow in human beings. AB - BACKGROUND: A number of studies have investigated vertebral artery flow velocity. Because perfusion relates to the volume of blood flowing through the vessel, this parameter is of great importance when vertebral artery hemodynamics are investigated. We could not find any such Doppler studies in the literature, possibly because of known errors with previous techniques. New advanced color coded duplex sonography has since been validated and may be used with confidence for volume flow investigations. OBJECTIVE: To use advanced color-coded duplex sonography to investigate volume flow through the vertebral arteries during cervical rotation, as well as before and after spinal manipulation therapy. DESIGN AND SETTING: A randomized controlled study at a university hospital vascular laboratory. PARTICIPANTS: Twenty university students. RESULTS: Volume blood flow through the vertebral arteries does not change with cervical rotation or after spinal manipulation therapy. CONCLUSION: This appears to be the first in vivo Doppler study on human vertebral artery volume blood flow. Our results indicate that in symptom-free subjects there is no change in vertebral artery perfusion during rotation in spite of significant changes in flow velocity. This finding, as well as the observed changes in flow velocity reported by others, may be explained by a positional change in the vertebral artery diameter. In addition, we have investigated volume blood flow in the vertebral arteries before or after spinal manipulation therapy but found no significant changes. PMID- 10478768 TI - Effect of premanipulative tests on vertebral artery and internal carotid artery blood flow: a pilot study. AB - BACKGROUND: Neck manipulation occasionally causes stroke after trauma to the vertebral or internal carotid artery. Premanipulativ e tests involving cervical spine rotation or extension have been recommended to detect patients at risk of neurovascular ischemia. However, the effect of these procedures on extracranial blood flow is not well established, and their validity is thus controversial. OBJECTIVE: To determine the effect of premanipulative tests involving cervical spine rotation or extension on vertebral artery and internal carotid artery blood flow parameters. DESIGN: Two-group experimental study. SUBJECTS: Twenty subjects consisting of 16 patients treated with physiotherapy and four volunteers. METHODS: Subjects were tested with a recommended premanipulative protocol by both an independent physiotherapist and an investigator. One group consisted of 10 subjects with signs or symptoms indicative of neurovascular ischemia on premanipulative testing, with 10 subjects with no signs or symptoms indicative of neurovascular ischemia on premanipulative testing comprising the second group. Hemodynamic measurements for both vertebral and both internal carotid arteries were taken by use of duplex Doppler ultrasonography with color-flow imaging with the subjects in the following positions: neutral, end-range extension, 45 degrees contralateral rotation, end-range contralateral rotation, and combined end-range contralateral rotation/extension. RESULTS: The reliability of premanipulative testing was supported. Significant changes in flow velocity of the vertebral artery (and to a lesser extent of the internal carotid artery) were shown in end range positions involving rotation and extension. No meaningful significant differences were found between the two groups. CONCLUSIONS: Screening procedures that use rotation and extension may be useful tests of the adequacy of collateral circulation. A larger study is needed to determine whether subjects testing positive significantly differ from those testing negative. PMID- 10478769 TI - Chronic spinal pain syndromes: a clinical pilot trial comparing acupuncture, a nonsteroidal anti-inflammatory drug, and spinal manipulation. AB - OBJECTIVE: To compare needle acupuncture, medication (tenoxicam with ranitidine), and spinal manipulation for managing chronic (>13 weeks duration) spinal pain syndromes. DESIGN: Prospective, randomized, independently assessed preintervention and postintervention clinical pilot trial. SETTING: Specialized spinal pain syndrome out-patient unit at Townsville General Hospital, Queensland, Australia. SUBJECTS: Seventy-seven patients (without contraindication to manipulation or medication) were recruited. INTERVENTIONS: One of three separate, clearly defined intervention protocols: needle acupuncture, nonsteroidal anti inflammatory medication, or chiropractic spinal manipulation. MAIN OUTCOME MEASURES: Main outcome measures were changes (4 weeks vs. initial visit) in the scores of the (1) Oswestry Back Pain Disability Index, (2) Neck Disability Index, and (3) three visual analogue scales of local pain intensity. RESULTS: Randomization was successful. After a median intervention period of 30 days, spinal manipulation was the only intervention that achieved statistically significant improvements (all expressed as percentages of the original scores) with (1) a reduction of 30.7% on the Oswestry scale, (2) an improvement of 25% on the neck disability index, and (3) reductions on the visual analogue scale of 50% for low back pain, 46% for upper back pain, and 33% for neck pain (all P<.001). Neither of the other interventions showed any significant improvement on any of the outcome measures. CONCLUSIONS: The consistency of the results provides, in spite of several discussed shortcomings of this pilot study, evidence that in patients with chronic spinal pain syndromes spinal manipulation, if not contraindicated, results in greater improvement than acupuncture and medicine. PMID- 10478770 TI - Preliminary study of the reliability of assessment procedures for indications for chiropractic adjustments of the lumbar spine. AB - OBJECTIVE: To assess the intraexaminer and interexaminer reliability of clinicians trained in flexion-distraction technique to determine the need for chiropractic adjustment of each segment of the lumbar spine. DESIGN: This was an intraexaminer and interexaminer reliability study of commonly used chiropractic assessment procedures, including static and motion palpation and visual observation. SETTING: Chiropractic college; by four licensed chiropractors trained in flexion-distraction technique, two with more than 20 years' experience and two with 3 or fewer years' experience. SUBJECTS: Subjects were 18 volunteers; 16 were symptom free, and 2 had low back pain at the time the study was conducted. MAIN OUTCOME MEASURE: The kappa statistic was computed for all comparisons and interpreted in categories ranging from "poor" (<0.00) to "almost perfect" (>0.80). RESULTS: Intraexaminer reliability was greater than interexaminer reliability. For intraexaminer reliability there was considerable variation by segment and among the four examiners, but intraexaminer reliability appeared generally higher than interexaminer reliability. Overall, more subluxations were identified on the second examination than on the first. For interexaminer reliability, kappa scores were generally in the "poor" to "slight" categories. DISCUSSION: The results of this study, similar to those of other studies, indicate that even chiropractors trained in the same technique seem to show little consensus on the indications for the necessity to adjust specific segments of the spine. A more standardized assessment approach might be helpful in improving the reliability of diagnostic assessments. PMID- 10478771 TI - Prevalence of hyperplastic articular pillars in the cervical spine and relationship with cervical lordosis. AB - BACKGROUND: Cervical lordosis is often used as an indicator for a number of clinical conditions ranging from traumatic to degenerative. Previous research has indicated that a number of factors may change the lordosis. However, the link between hyperplastic articular pillars and cervical lordosis measurements has never been studied. OBJECTIVE: To investigate the reliability of determining articular pillar hyperplasia, to determine its prevalence, and to compare lordosis measurements between persons with and without hyperplasia of the cervical articular pillars. METHODS: Twelve normal neutral lateral cervical radiographs were chosen for the reliability study. Two chiropractors independently evaluated the articular pillars of C3 through C6 by drawing lines along the planes of the superior and inferior articular surfaces of each pillar. Each pillar was categorized as "normal" or "hyperplastic" depending on the convergence or divergence of these lines. One examiner repeated this procedure after a 1-month interval. Percent agreement and kappa statistics were calculated for interexaminer and intraexaminer agreement. Forty-eight normal neutral lateral cervical radiographs with a horizontal Chamberlain's line were evaluated for the presence or absence of pillar hyperplasia. Two measurement techniques were used to assess the cervical lordosis: the method of Jochumsen and the "angle of the cervical lordosis." The unpaired t test was used to compare the angle of cervical lordosis measurements between the two groups. The Mann-Whitney U test was used to analyze data obtained with the method of Jochumsen. The prevalence of pillarhyperplasia was calculated on 100 normal lateral cervical radiographs. RESULTS: Interexaminer and intraexaminer reliability of determining pillar hyperplasia was fair to substantial (kappa = 0.4 to 0.61; 75% to 92%). Forty-six percent of the patients demonstrated pillar hyperplasia of at least one cervical level. Patients with pillar hyperplasia had a mean cervical curve of 31.52 degrees, whereas the curve in patients without pillar hyperplasia was 44.76 degrees (P = .0001). The method of Jochumsen also detected a significant difference in the curve measurements between the two patient groups (P = .0127). CONCLUSION: Articular pillar hyperplasia is common and significantly reduces the cervical lordosis measurements. Accepted normal values for lordosis evaluation need to be reassessed and articular pillar configuration considered when treatment plans include attempts to restore a "normal" cervical lordosis. Previous studies attempting to link lordosis measurements with pathologic conditions or symptoms may need to be reevaluated. PMID- 10478772 TI - Stress: the chiropractic patients' self-perceptions. AB - BACKGROUND: Psychosocial stress pervades modern life and is known to have an impact on health. Pain, especially chronic back pain, is influenced by stress. Various strategies have been shown to successfully reduce stress and its consequences. OBJECTIVES: This study explores stress as a potential disease trigger among chiropractic patients. METHOD: A descriptive study was undertaken to ascertain the stress perceptions of chiropractic patients. Purposive sampling of chiropractic practices and convenience sampling of patients was undertaken. Patients were allocated to 1 of 4 groups according to their presentation: acute, chronic biomechanical, fibromyalgia, or maintenance care. Participating patients were requested to complete a questionnaire. RESULTS: Of the 138 patients attending 1 of 10 participating chiropractic clinics, more than 30% regarded themselves as moderately to severely stressed, and over 50% felt that stress had a moderate or greater effect on their current problem. Some 71% of patients felt it would be helpful if their chiropractic care included strategies to help them cope with stress, and 44% were interested in taking a self-development program to enhance their stress management skills. CONCLUSION: Patient perceptions are known to be important in health care. A number of chiropractic patients perceive they are moderately or severely stressed. Interventions that reduce stress, or even the patient's perception of being stressed, may be construed as valid, non specific clinical interventions. It may be timely for chiropractors to actively contemplate including stress management routinely in their clinical care protocols. PMID- 10478774 TI - Chiropractic treatment of coccygodynia via instrumental adjusting procedures using activator methods chiropractic technique. AB - OBJECTIVE: To discuss a case of coccygodynia that responded favorably to conservative chiropractic adjusting procedures with the Activator Methods Chiropractic Technique (AMCT) and the Activator II Adjusting Instrument (AAI II). CLINICAL FEATURES: A 29-year-old woman had unremitting coccygeal pain of 3 weeks' duration. The problem began after she had moved heavy boxes while at work. The pain was characterized by a continual dull ache in the coccygeal region, accompanied by intermittent sharp pain, particularly upon sitting or rising from a seated position. She had been taking self-prescribed over-the-counter analgesics (aspirin and ibuprofen) for 3 weeks without obtaining relief. INTERVENTION AND OUTCOME: Treatment consisted of mechanical force, manually assisted, short-lever (MFMA) chiropractic adjusting procedures to the coccygeal area, primarily the sacrococcygeal ligament. The AAI II was used to deliver the adjustment according to diagnostic and treatment protocol specified for AMCT. The patient experienced first treatment. CONCLUSION: Chiropractic coccygeal manipulation may be effectively delivered via instrumental adjustment in certain cases of coccygodynia. The use of an AAI II in administering the coccygeal adjustment has the benefit of being a gentle, noninvasive procedure, as well as being comfortably tolerated by the patient. This method of coccygeal adjustment may bear consideration in certain cases of coccygodynia. PMID- 10478773 TI - A review of biomechanics of the central nervous system--Part III: spinal cord stresses from postural loads and their neurologic effects. AB - OBJECTIVE: To review literature pertaining to neurologic disorders stemming from abnormal postures of the spine. DATA COLLECTION: A hand search of available reference texts and a computer search of literature from Index Medicus sources was performed, with special emphasis placed on spinal cord stresses and strains caused by various postural rotations and translations of the skull, thorax, and pelvis. RESULTS: Spinal postures will often deform the neural elements within the spinal canal. Spinal postures can be broken down into four types of loading: axial, pure bending, torsion, and transverse, which cause normal and shear stresses and strains in the neural tissues and blood vessels. Prolonged stresses and strains in the neural elements cause a multitude of disease processes. CONCLUSION: Four types of postural loads create a variety of stresses and strains in the neural tissue, depending on the exact magnitude and direction of the forces. Transverse loading is the most complex load. The stresses and strains in the neural elements and vascular supply are directly related to the function of the sensory, motor, and autonomic nervous systems. The literature indicates that prolonged loading of the neural tissue may lead to a wide variety of degenerative disorders or symptoms. The most offensive postural loading of the central nervous system and related structures occurs in any procedure or position requiring spinal flexion. Thus flexion traction, rehabilitation positions, exercises, spinal manipulation, and surgical fusions in any position other than lordosis for the cervical and lumbar spines should be questioned. PMID- 10478775 TI - Extraspinal enthesopathy caused by isotretinoin therapy. AB - OBJECTIVE: To discuss a case of diffuse peripheral enthesopathy in a patient previously treated with long-term isotretinoin (Accutane) for severe acne. CLINICAL FEATURES: A 47-year old man with 1 month history of moderate neck and right upper extremity pain, with hypoesthesia of the right second and third fingers. Palpable bony prominences around multiple superficial joints were noted on physical examination, raising the initial question of osteochondromatosis. Multiple active acne pustules were noted. A limited skeletal survey demonstrated diffuse peripheral enthesophyte formation and hyperostoses, resembling those of diffuse idiopathic skeletal hyperostosis, but without accompanying spinal changes. A history of long-term Accutane therapy was then elicited. INTERVENTION AND OUTCOME: The enthesopathy was believed to represent an asymptomatic, longstanding, iatrogenically induced abnormality. No specific therapy or follow up was indicated. The patient had discontinued use of Accutane years ago. Cervical symptoms improved with four sessions of cervical traction and nonsteroidal anti-inflammatory medications, but upper extremity symptoms were refractory. CONCLUSION: Accutane-induced enthesopathy should be considered in individuals with correlating radiologic and clinical features and history of retinoic acid therapy for acne. This should be a diagnosis by exclusion, after eliminating other potential causes of peripheral enthesopathy, particularly diffuse idiopathic skeletal hyperostosis, seronegative spondylarthropathy, and fluorosis. PMID- 10478776 TI - Possible manifestation of temporomandibular joint dysfunction on chiropractic cervical X-ray studies. PMID- 10478777 TI - Schizophrenia and complications of pregnancy and labor: an individual patient data meta-analysis. AB - Several epidemiological studies have reported an association between complications of pregnancy and delivery and schizophrenia, but none have had sufficient power to examine specific complications that, individually, are of low prevalence. We, therefore, performed an individual patient meta-analysis using the raw data from case control studies that used the Lewis-Murray scale. Data were obtained from 12 studies on 700 schizophrenia subjects and 835 controls. There were significant associations between schizophrenia and premature rupture of membranes, gestational age shorter than 37 weeks, and use of resuscitation or incubator. There were associations of borderline significance between schizophrenia and birthweight lower than 2,500 g and forceps delivery. There was no significant interaction between these complications and sex. We conclude that some abnormalities of pregnancy and delivery may be associated with development of schizophrenia. The pathophysiology may involve hypoxia and so future studies should focus on the accurate measurement of this exposure. PMID- 10478778 TI - Does preexisting abnormality cause labor-delivery complications in fetuses who will develop schizophrenia? AB - Many authors have suggested that theoretically the labor-delivery complications (LDCs) that frequently appear in the histories of individuals with schizophrenia represent the secondary consequence of preexistent abnormality in the fetus. The question of whether LDCs are systematically associated with prenatal complications and fetal abnormality was studied in 70 singleton schizophrenia patients, in 23 monozygotic twin pairs discordant and 10 pairs concordant for schizophrenia, and in 33 individuals with inferred genetic risk for schizophrenia. Schizophrenia cases with signs of prenatal abnormality (reduced head size, increased minor physical anomalies, greater within-twin-pair birthweight differences) did not have more LDCs than other schizophrenia cases. LDCs were not more frequent in genetic-risk cases with congenital malformations than in genetic-risk cases without malformations. Instead, individuals with schizophrenia who had a history of abnormal length of labor had significantly fewer pregnancy complications and minor physical anomalies than did other individuals with schizophrenia. No support was found for suggestions that LDCs among individuals who have not yet developed schizophrenia are the result of identifiable preexistent fetal abnormality. PMID- 10478779 TI - Dispelling the stigma of schizophrenia: II. The impact of information on dangerousness. AB - This study addressed a relatively neglected topic in schizophrenia: identifying methods to reduce stigma directed toward individuals with this disorder. The study investigated whether presentation of information describing the association between violent behavior and schizophrenia could affect subjects' impressions of the dangerousness of both a target person with schizophrenia and individuals with mental illness in general. Subjects with and without previous contact with individuals with a mental illness were administered one of four "information sheets" with varying information about schizophrenia and its association with violent behavior. Subjects then read a brief vignette of a male or female target individual with schizophrenia. Results showed that subjects who reported previous contact with individuals with a mental illness rated the male target individual and individuals with mental illness in general as less dangerous than did subjects without previous contact. Subjects who received information summarizing the prevalence rates of violent behavior among individuals with schizophrenia and other psychiatric disorders (e.g., substance abuse) rated individuals with a mental illness as less dangerous than did subjects who did not receive this information. Implications of the findings for public education are discussed. PMID- 10478780 TI - Changing attitudes about schizophrenia. AB - Research on the effectiveness of short-term education programs in changing societal attitudes about mental illness has been mixed. Education efforts seem to be mediated by characteristics of the program participants. This study determines whether the effects of a specially prepared, semester-long course on severe mental illness are mediated by pre-education knowledge about and contact with severe mental illness. Eighty-three participants who were enrolled in either a course on severe mental illness or general psychology completed the Opinions about Mental Illness Questionnaire before beginning the course and at completion. Research participants also completed a pre-and posttest of knowledge about mental illness and a pretest on their contact with people who have severe mental illness. The education program had positive effects on some attitudes about mental illness. Interestingly, the effects of education group interacted with pre education knowledge and contact and varied depending on attitude. Participants with more pre-education knowledge and contact were less likely to endorse benevolence attitudes after completing the education program. Participants with more intimate contact showed less improvement in attitudes about social restrictiveness. Implications of these augmentation and ceiling effects are discussed. PMID- 10478781 TI - Treatment-resistant schizophrenia and staff rejection. AB - This study examined the relationship between characteristics of patients suffering from treatment-refractory schizophrenia and staff rejection and criticism. Subjects were 30 inpatients with treatment-resistant schizophrenia and the 29 staff members treating them. Measures included assessment of the patients' symptoms and aggression risk profile using the Positive and Negative Syndrome Scale (PANSS) and assessment of staff attitudes toward these patients using the Patient Rejection Scale (PRS). Nursing staff completed the Nurses' Observation Scale for Inpatient Evaluation (NOSIE). PRS ratings did not correlate with patients' demographic and treatment characteristics. Significant correlations existed, however, between increased staff rejection and higher scores for PANSS cognitive factor and NOSIE manifest psychosis factor. Negative symptoms, although preponderant in the patient sample, were not significant predictors of staff rejection on the PRS. Older nursing staff tended to view patients as more irritable and manifestly psychotic. These findings suggest that disorganized behavior and impaired cognition dysfunction areas are more likely to be associated with high levels of rejection among staff working with treatment resistant schizophrenia patients. Incorporation of the relatively new concepts of cognitive dysfunction and treatment resistance in staff training programs and multidisciplinary team reviews may greatly benefit schizophrenia patients and the staff treating them. PMID- 10478782 TI - Mental health consumers' experience of stigma. AB - The extent to which mental health consumers encounter stigma in their daily lives is a matter of substantial importance for their recovery and quality of life. This article summarizes the results of a nationwide survey of 1,301 mental health consumers concerning their experience of stigma and discrimination. Survey results and followup interviews with 100 respondents revealed experience of stigma from a variety of sources, including communities, families, churches, coworkers, and mental health caregivers. The majority of respondents tended to try to conceal their disorders and worried a great deal that others would find out about their psychiatric status and treat them unfavorably. They reported discouragement, hurt, anger, and lowered self-esteem as results of their experiences, and they urged public education as a means for reducing stigma. Some reported that involvement in advocacy and speaking out when stigma and discrimination were encountered helped them to cope with stigma. Limitations to generalization of results include the self-selection, relatively high functioning of participants, and respondent connections to a specific advocacy organization the National Alliance for the Mentally Ill. PMID- 10478783 TI - Effects of the label "schizophrenia" on causal attributions of violence. AB - We investigated the relation between the label of "schizophrenia" and causal attributions of violence. Undergraduates read 1 of 10 scenarios in which two variables were manipulated: a psychiatric label and environmental stress. The scenario described an employee who acted violently toward his boss. Subjects made causal attributions for the employee's behavior by completing an adapted version of the Causal Dimension Scale II. Subjects also completed a questionnaire designed to explore several issues concerning the effects of the schizophrenia label on perceptions of behavior. Contrary to the primary hypothesis, the schizophrenia label did not lead subjects to make significantly more personality causal attributions for violent behavior. With increasing environmental stress, subjects did make significantly fewer personality attributions. A follow-up study using practicing clinicians as subjects yielded similar findings. The results of these studies are discussed in light of perceived stereotypes of persons with schizophrenia and conceptual issues in attribution research. PMID- 10478784 TI - Violence in inpatients with schizophrenia: a prospective study. AB - Accurate evaluations of the dangers posed by psychiatric inpatients are necessary, although a number of studies have questioned the accuracy of violence prediction. In this prospective study, we evaluated several variables in the prediction of violence in 63 inpatients with a DSM-IV diagnosis of schizophrenia or schizoaffective disorder. Nurses rated violent incidents with the Overt Aggression Scale. During hospitalization, sociodemographic variables, clinical history, neurological soft signs, community alcohol or drug abuse, and electroencephalographic abnormalities did not differ between violent and nonviolent groups. Violent patients had significantly more positive symptoms as measured by the Positive and Negative Syndrome Scale (PANSS), higher scores on the PANSS general psychopathology scale, and less insight in the different constructs assessed. A logistic regression was performed to discriminate between violent and nonviolent patients. Three variables entered the model: insight into symptoms, PANSS general psychopathology score, and violence in the previous week. The actuarial model correctly classified 84.13 percent of the sample; this result is significantly better than chance for the base rate of violence in this study. At hospital admission, clinical rather than sociodemographic variables were more predictive of violence. This finding has practical importance because clinical symptoms are amenable to therapeutic approaches. This study is the first to demonstrate that insight into psychotic symptoms is a predictor of violence in inpatients with schizophrenia. PMID- 10478785 TI - Course of violence in patients with schizophrenia: relationship to clinical symptoms. AB - To understand the heterogeneity of violent behaviors in patients with schizophrenia, one must consider underlying clinical symptoms of the illness and their change over time. The purpose of this study was to examine persistence and resolution of violence in relation to psychotic symptoms, ward behaviors, and neurological impairment. Psychiatric symptoms and ward behaviors were assessed in violent inpatients with schizophrenia or schizoaffective disorder and in nonviolent controls on entry into the study. Patients were followed for 4 weeks; those who showed resolution of assaults over this time were classified as transiently violent, and those who remained assaultive were categorized as persistently violent. At the end of the 4 weeks, psychiatric symptoms, ward behaviors, and neurological impairment were assessed. Overall, the two violent groups presented with more severe psychiatric symptoms and were judged to be more irritable than the nonviolent control subjects, but the transiently violent patients showed improvement in symptoms over time. At the end of 4 weeks, the persistently violent patients had evidence of more severe neurological impairment, hostility, suspiciousness, and irritability than the other two groups. Canonical discriminant analyses identified two significant dimensions differentiated the groups. The first, characterized by positive psychotic symptoms, differentiated the violent patients from the control subjects; the second, characterized by neurological impairment and high endpoint score for negative symptoms, differentiated the transiently from the persistently violent patients. Identification of certain symptoms associated with different forms of violence has important implications for the prediction and differential treatment of violent behavior in patients with schizophrenia. PMID- 10478786 TI - Life events and completed suicide in schizophrenia: a comparison of suicide victims with and without schizophrenia. AB - Adverse life events are an established risk factor in completed suicide. However, few studies have examined life events and suicide in schizophrenia. We investigated and compared schizophrenia suicide victims and age- and sex-matched victims without schizophrenia as part of a psychological autopsy study of all suicides in Finland over a 12-month period. Recent life events were examined retrospectively by interviewing next of kin using a structured life event questionnaire. Overall, nearly half (46%) the schizophrenia subjects had had adverse life events before suicide, significantly less than the nonschizophrenia subjects (83%). In both groups, however, suicide was preceded by life events independent of the victims' own behavior, such as death of a close person or illness in the family. Life events overall were more common among schizophrenia outpatients (52%) than inpatients (22%), and the association of life events with suicide was clearest among a subgroup of outpatients in residual phase who had used neuroleptic medication regularly. Overall, the prevalence of recent adverse life events varied between clinical subgroups of victims with schizophrenia, which may have implications for suicide prevention. PMID- 10478787 TI - Suicide risk in schizophrenia: an analysis of 17 consecutive suicides. AB - The aim of this study was to investigate interactional factors related to the recognition of suicide risk in patients with schizophrenia. The study focused on 17 schizophrenia patients who had committed suicide during the National Suicide Prevention Project in Finland between April 1, 1987, and March 31, 1988, in the province of Kuopio. Consensus case reports were assembled by using the psychological autopsy method. Study methods included structured and in-depth interviews of next of kin and interviews of health care or social services workers who had treated the suicide victims. Male and female patients with schizophrenia committed suicide in equal proportions. Most had suffered from schizophrenia for more than 15 years; all but one had been receiving psychiatric treatment at the time of suicide. Retrospective assessment indicated that 59 percent of the patients were clinically depressed at the time of suicide. In 76 percent of the cases, the mental health professionals involved in treatment had not believed that there was a risk of suicide during their last contact with the patient. In 29 percent of the cases, the patient's paranoid ideas concerning treatment personnel had increased. Patients' withdrawal from human relationships because of depression was related to loss of the treatment professionals' concern for the patients. The findings in this descriptive study suggest that withdrawal by a patient with schizophrenia and an increase in the patient's paranoid behavior should be regarded as signals of risk of suicide. PMID- 10478788 TI - Deinstitutionalization and schizophrenia in Finland II: discharged patients and their psychosocial functioning. AB - Three representative cohorts of schizophrenia patients deinstitutionalized from psychiatric hospitals in 1982, 1986, and 1990 were followed up for 3 years in Finland. Patients of the last cohort were older, more disturbed, and had been ill for a longer time than those discharged at the beginning of the 1980s. Despite this, the mortality of patients deinstitutionalized in 1990 did not increase, and their psychosocial functioning seemed to become even better during the 3-year follow-up period compared with those deinstitutionalized during the previous decade. Patients who had been discharged in 1990 were more often living alone than those discharged in the 1980s. Homelessness was rare throughout the study period. In general, patients were more satisfied with their life situation at follow-up compared with that on discharge. Furthermore, most patients were satisfied with their treatment situation. Altogether, the psychiatric care system seemed to be able to meet schizophrenia patients' need for care fairly well during the rapid deinstitutionalization process in Finland. More attention, however, should be paid to the loneliness and social withdrawal of discharged patients as well as to other disabilities in their social functioning. PMID- 10478789 TI - Measurement of delusional ideation in the normal population: introducing the PDI (Peters et al. Delusions Inventory). AB - The Peters et al. Delusions Inventory (PDI) was designed to measure delusional ideation in the normal population, using the Present State Examination as a template. The multidimensionality of delusions was incorporated by assessing measures of distress, preoccupation, and conviction. Individual items were endorsed by one in four adults on average. No sex differences were found, and an inverse relationship with age was obtained. Good internal consistency was found, and its concurrent validity was confirmed by the percentages of common variance with three scales measuring schizotypy, magical ideation, and delusions. PDI scores up to 1 year later remained consistent, establishing its test-retest reliability. Psychotic inpatients had significantly higher scores, establishing its criterion validity. The ranges of scores between the normal and deluded groups overlapped considerably, consistent with the continuity view of psychosis. The two samples were differentiated by their ratings on the distress, preoccupation, and conviction scales, confirming the necessity for a multidimensional analysis of delusional thinking. Possible avenues of research using this scale and its clinical utility are highlighted. PMID- 10478790 TI - Structured Interview for Assessing Perceptual Anomalies (SIAPA). AB - Clinical descriptions of perceptual and attentional anomalies in schizophrenia emphasize phenomena such as flooding, or inundation, by sensory stimuli. A failure of sensory "gating" mechanisms in the brain is hypothesized to account for these symptoms, and this hypothesis has led to a marked interest in their putative psychophysiological substrates. However, there are no systematic analyses of the phenomenology of these perceptual experiences, nor has the hypothesized connection between the clinical phenomena and their reported psychophysiological substrates been tested. In this investigation, a structured interview instrument was developed to measure perceptual anomalies as distinct from hallucinations and to determine their prevalence across sensory modalities in schizophrenia in 67 schizophrenia subjects and 98 normal controls. The instrument includes Likert ratings of hypersensitivity, inundation, and selective attention to external sensory stimuli. Good interrater agreement, determined from interviews, was obtained. Schizophrenia subjects had significantly higher auditory, visual, and combined scores (i.e., across all modalities) than normal controls did, indicating significantly more perceptual anomalies. For the schizophrenia group, the prevalence of auditory and visual anomalies was significantly greater than the other sensory modalities. The data indicate that the putative phenomenological correlates of sensory gating may be reliably measured and tested with the Structured Interview for Assessing Perceptual Anomalies. PMID- 10478791 TI - Category learning and perceptual categorization in schizophrenia. AB - The aim of this study was to evaluate category learning in schizophrenia on tests of perceptual abstraction. Participants learned to categorize simple geometrical shapes. The categories were either well-defined (discrete categories, or DCs) or ill-defined (graded categories, or GCs). In DCs, the cues defining category membership can be verbalized in an all-or-none fashion, while in GCs they cannot be defined unambiguously. Three types of learning were used successively: serial presentation of category-exemplars, verbal description, and feedback. After the serial presentation, schizophrenia patients showed a deficit for GCs (p<0.005) but not for DCs (p = 0.98). After the verbal definition of GCs, the difference between schizophrenia patients and controls diminished (p = 0.09). Finally, after the feedback learning of GCs, a significant difference was observed again (p<0.0001), suggesting that schizophrenia patients were impaired in this learning paradigm. The GC-learning impairment after the serial presentation displayed a relationship with the score of the cognitive component assessed with the Positive and Negative Syndrome Scale (r = -0.66). In conclusion, these results suggest that the perceptual stage of abstraction is impaired in schizophrenia. This impairment can be partially compensated by instructions via top-down verbal processes. PMID- 10478792 TI - Altered parallel auditory processing in schizophrenia patients. AB - Patients with schizophrenia have impaired auditory processing that has been demonstrated by diminished P50 response to paired auditory stimuli in event related potential (ERP) studies. Cerebral processing can also be studied with magnetoencephalography (MEG). With a whole-head MEG, which enables one to simultaneously measure brain activity in both hemispheres, we investigated whether early parallel auditory processing is impaired in schizophrenia. Sequences of tones were monaurally presented to schizophrenia patients and healthy controls in a passive condition, and the event-related magnetic fields were recorded simultaneously over both auditory cortices. The interhemispheric latency difference of the P50m, but not that of the N100m, was significantly shorter in the patient group in the right-ear but not in the left-ear stimulus condition. Further, the ipsilateral P50m was significantly earlier in schizophrenia patients in the right-ear condition. This result suggests that schizophrenia affects the consecutive preconscious auditory processing in a different manner. PMID- 10478793 TI - Genetic variants of dopamine receptor D4 and psychopathology. AB - There is much evidence to indicate that the dopamine receptor D4 (DRD4) gene is involved in psychiatric disorders. We investigated the correlation between DRD4 gene polymorphism and the psychopathology of major psychoses, independently of diagnoses. Some 461 inpatients affected by major psychoses were assessed by the Operational Criteria checklist for psychotic illness and typed for DRD4 variants. The four symptomatologic factors-mania, depression, delusion, and disorganization were used as phenotype definitions. DRD4 Exon 3 long allele variants were associated with high delusional scores, with the most significant difference between alleles 2 and 7 (p = 0.004). DRD4 variants may, therefore, constitute a liability factor for development of delusional symptomatology in patients with major psychoses. PMID- 10478794 TI - First person account: family psychoeducation can change lives. PMID- 10478795 TI - Factors in serum regulate muscle development in P19 cells. PMID- 10478796 TI - Development of a serum-free medium for dihydrofolate reductase-deficient Chinese hamster ovary cells (DG44) using a statistical design: beneficial effect of weaning of cells. AB - To develop serum-free (SF) medium for dihydrofolate reductase-deficient Chinese hamster ovary cells (DG44), a statistical optimization approach based on a Plackett-Burman design was adopted. DG44 cells which were normally maintained in 10 serum medium were gradually weaned to 0.5% serum medium to increase the probability of successful growth in SF medium. A basal medium was prepared by supplementing Dulbecco's modified Eagle's medium and Ham's nutrient mixture F12 with hypoxanthine (10 mg/l) and thymidine (10 mg/l). Twenty-eight different supplements were selected as variables on the basis of their growth-promoting abilities. From statistical analysis, leucine, tryptophan, lysine, proline, histidine, hydrocortisone, ethanolamine, and phosphatidylcholine were identified as important components showing positive effects on cell growth. A new SF medium (SF-DG44) was formulated by supplementing the basal medium with these components. When the weaned cells were inoculated at 1.0 x 10(5) cells/ml, a maximum viable cell concentration of 6.4 x 10(3)) cells/ ml was achieved in SF-DG44 medium. In contrast, when the unweaned cells were used, a concentration of only 4.1 x 10(5) cells/ml was reached under the same culture conditions, indicating that weaning of cells improves cell growth in SF medium. In summary, we found that development of a novel SF medium for DG44 cells was facilitated using a Plackett-Burman design technique and weaning of cells. PMID- 10478797 TI - ECC-1 human endometrial cells as a model system to study dioxin disruption of steroid hormone function. AB - ECC-1, an established epithelial cell line derived from an adenocarcinoma of human endometrial lining, was examined for growth optimization, steroid hormone receptor- and Ah receptor content, and dioxin modulation of estrogen receptor function. Proliferation of ECC-1 cells was accelerated by growth on a lethally irradiated feeder layer of murine 3T3 fibroblasts. Immunoblot analysis demonstrated the presence of Ah receptor an intracellular protein that binds and regulates the toxic action of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The Ah receptor was functional in these cells as assessed by concentration and kinetic patterns of CYP1A1-mediated 7-ethoxycoumarin O-deethylase (ECOD) induction. The half-maximal effective concentration (EC50) for TCDD was 0.2 nM, and maximal activity appeared after 24-h exposure. A limited structure-activity examination of ECOD activity provided additional evidence for Ah receptor involvement. Competitive binding assays were performed to examine kinetic parameters for estrogen, progesterone, and glucocorticoid receptors. Binding parameters of dissociation constant (Kd) and number of binding sites (Bmax) derived from Scatchard analysis were: estrogen, Kd = 0.67 nM; Bmax = 321 fmol/mg cytosolic protein; progesterone, Kd = 1.31 nM; Bmax = 258 fmol/mg cytosolic protein; dexamethasone, Kd = 1.75 nM, Bmax = 128 fmol/mg cytosolic protein. Exposure of ECC-1 cells to TCDD reduced the estrogen receptor level by 40% without affecting the Kd value, and reduced estrogen receptor-mediated transcription by 50% assessed by transient transfection of an estrogen-responsive reporter plasmid. These data suggest that the ECC-1 cell line is a useful model system for examining the action of dioxin in human endometrial tissue. Both the estrogen receptor and Ah receptor have been implicated in diseases of the endometrium, and examining their interactions may elucidate mechanisms of uterine disease etiology, as well as potential targets for disease prevention. PMID- 10478798 TI - Isolation and characterization of a human hepatic epithelial-like cell line (AKN 1) from a normal liver. AB - The isolation and characterization of human liver cell lines are rather difficult due to limited material and poor growth in cell culture. In this report, we present the isolation, culture and characterization of a new epithelial-like liver cell line (AKN-1) with a heterogeneous cell population and many characteristics of the biliary epithelium. The AKN-1 cell line stained positively with antibodies to epithelial cytokeratin polypetides CK 8, 18, and 19. In addition, the cell line expressed the anti-human epithelial-related antigen (MOC 31), the human epithelial antigen (HEA), and the gamma-glutamyl transpeptidase, the hematopoietic growth factor, stem cell factor, and also its receptor, c-kit. The cell line failed to express albumin and factor 8 by immunohistochemistry. It did show, however, a twofold increase in 7-ethoxyresorufin-O-deethylase activity. Cytogenetic characterization revealed rare breakpoints in chromosome 2, which to our knowledge, have not yet been reported in liver cells. PMID- 10478799 TI - Characterization of immortalized rabbit lacrimal gland epithelial cells. AB - To establish an immortalized lacrimal gland epithelial cell line, the orbital lacrimal glands of normal New Zealand White rabbits were multiply injected with an immortalizing amphotropic retroviral vector (LXSN16E6E7) containing the E6 and E7 genes of human papillomavirus type 16. Lacrimal glands were removed after 2 d and acinar epithelial cells were isolated and cultured on Matrigel-coated 60 mm2 plates containing DMEM-F12 supplemented with 5% Nu-serum V. Transformed cells were selected in G418 sulfate for 7 d and passaged. Morphology of the immortalized cells was similar to that described for normal acinar cells both in vivo and in vitro, with rough endoplasmic reticulum and secretory granules. These characteristics remained unchanged and the cells continued to exhibit typical polygonal epithelioid structure. The cells have been maintained in culture for 14 mo. and have gone through 58 passages without loss of proliferation or epithelial cell characteristics. Immunohistochemistry and Western blots showed positive reactivity to secretory component, transferrin, and transferrin receptor, which are typical proteins found in the lacrimal gland. Functional analysis by stimulation with a cholinergic agonist, carbachol (100 microM), resulted in a significant release of protein. This is the first report of an immortalized rabbit lacrimal epithelial cell. These cells will provide a valuable tool for the molecular analysis of lacrimal gland epithelial cell functions. PMID- 10478801 TI - Nitric oxide donors, sodium nitroprusside and S-nitroso-N-acetylpencillamine, stimulate myoblast proliferation in vitro. AB - Nitric oxide (NO) is an inter- and intracellular messenger involved in a variety of physiologic and pathophysiologic conditions. The effect of two NO donors, sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP) and their effect on myoblast proliferation was examined. Both donors stimulated an increase in myoblast cell number over a range (1-10 microM) of donor concentrations. However, 50 microM SNAP inhibited myoblast proliferation. Cell numbers from cultures treated with degraded 10 microM SNAP were equivalent to the control. Therefore, it appears NO can stimulate as well as inhibit myoblast proliferation. PMID- 10478800 TI - H-ras-transformed NRK-52E renal epithelial cells have altered growth, morphology, and cytoskeletal structure that correlates with renal cell carcinoma in vivo. AB - We studied the effect of the ras oncogene on the growth kinetics, morphology, cytoskeletal structure, and tumorigenicity of the widely used NRK-52E rat kidney epithelial cell line and two H-ras oncogene-transformed cell lines, H/1.2-NRK-52E (H/1.2) and H/6.1-NRK-52E (H/6.1). Population doubling times of NRK-52E, H/1.2, and H/6.1 cells were 28, 26, and 24 h, respectively, with the transformed cells reaching higher saturation densities than the parent cells. NRK-52E cells had typical epithelial morphology with growth in colonies. H/1.2 and H/6.1 cell colonies were more closely packed, highly condensed, and had increased plasma membrane ruffling compared to parent cell colonies. NRK-52E cells showed microfilament, microtubule, and intermediate filament networks typical of epithelial cells, while H/1.2 and H/6.1 cells showed altered cytoskeleton architecture, with decreased stress fibers and increased microtubule and intermediate filament staining at the microtubule organizing center. H/1.2 and H/6.1 cells proliferated in an in vitro soft agar transformation assay, indicating anchorage-independence, and rapidly formed tumors in vivo with characteristics of renal cell carcinoma, including mixed populations of sarcomatoid, granular, and clear cells. H/6.1 cells consistently showed more extensive alterations of growth kinetics, morphology, and cytoskeleton than H/1.2 cells, and formed tumors of a more aggressive phenotype. These data suggest that analysis of renal cell characteristics in vitro may have potential in predicting tumor behavior in vivo, and significantly contribute to the utility of these cell lines as in vitro models for examining renal epithelial cell biology and the role of the ras proto-oncogene in signal transduction involving the cytoskeleton. PMID- 10478802 TI - Differentiation markers of mouse C2C12 and rat L6 myogenic cell lines and the effect of the differentiation medium. AB - The differentiation grade of cells in culture is dependent on the composition of the culture medium. Two commonly used myogenic cell lines, mouse C2C12 and rat L6, usually differentiate at a low concentration of horse serum. In this study we compared the effect of horse serum with a medium containing a low percentage of Ultroser G and rat brain extract. The maturation grade was evaluated on the basis of various biochemical, (immuno)histochemical and cell-physiological parameters. Substitution of horse serum by Ultroser G and rat brain extract during the differentiation phase resulted in a higher maturation grade of the myotubes of both cell lines, on the basis of creatine kinase activity and the diameter of the myotubes. In addition, the C2C12 myotubes display cross-striation, contain a higher percentage of creatine kinase muscle-specific isoenzyme MM, show a ninefold increase in acetylcholine receptor (AChR) clusters, form a continuous basement membrane, and have a lower resting cytosolic Ca2+ concentration. L6 myotubes show a fivefold increase in AChR clusters and a twofold increase in the expression of the mRNA of the epsilon-subunit of AChR.C2C12 cells show spontaneous contraction and response of cytosolic Ca2+ to various stimulants in contrast to L6 cells which do not. These studies established that the Ultroser G/brain extract medium leads to a higher differentiation grade of both cell lines, but parameters appropriate for use as differentiation markers appear to differ between both cell lines. PMID- 10478804 TI - Amplification of the Y chromosome in three murine tumor cell lines transformed in vivo by different human prostate cancers. AB - Conventional and molecular cytogenetic analyses of three murine cancer cell lines that had been induced in male athymic mice by the injection of three different human prostate cancer cell lines revealed selective amplification of the Y chromosome. In particular, analysis of metaphase and interphase nuclei by fluorescence in situ hybridization (FISH) with the mouse Y chromosome-specific DNA painting probe revealed the presence of various numbers of Y chromosomes, ranging from one to eight, with a large majority of nuclei showing two copies (46.5-60.1%). In Interphase nuclei, the Y chromosomes showed distinct morphology, allowing identification irrespective of whether the preparations were treated for 15 min or for 5 h with Colcemid, a chemical known to cause chromosome condensation. However, FISH performed on human lymphocyte cultures with chromosome-specific DNA painting probes other than the Y chromosome did not reveal condensed chromosome morphology in interphase nuclei even after 12 h of Colcemid treatment. Our FISH results indicate that (1) the Y chromosome is selectively amplified in all three cell lines; (2) the mouse Y chromosome number is comparable in both interphase and metaphase cells: (3) the Y chromosome number varies between one and eight, with a large majority of cells showing two or three copies in most interphase nuclei; (4) the condensation of the Y chromosome is not affected by the duration of Colcemid treatment but by its inherent DNA constitution; and (5) the number of copies of the Y chromosome is increased and retained not only in human prostate tumor cell lines but also in murine tumors induced by these prostate tumor cell lines. PMID- 10478803 TI - Growth and differentiation regulate CD44 expression on human keratinocytes. AB - Several members of the CD44 family of hyaluronan receptors are expressed on keratinocytes. To identify factors that might be important in regulating CD44 expression, we studied CD44 expression on keratinocytes growing in vitro under a variety of conditions and on cells isolated directly from epidermis. Using Western immunoblots and metabolic labeling, we showed that the pattern of CD44 proteins expressed by keratinocytes was strongly influenced by growth and differentiation. Many protein forms of CD44 are expressed on proliferating keratinocytes in preconfluent cultures, whereas only a few forms are expressed on differentiated cells and in confluent cultures. In preconfluent monolayers, at least four splice variants were identified, including epican, CD44H, CD44E, and a 180-kDa variant. In differentiated cells or in confluent cultures, by contrast, only epican and the 180-kDa protein variant were found. Synthesis of all variants is strongly downregulated when keratinocytes become confluent or when they differentiate. Epican is the predominant form of CD44 on keratinocytes under all conditions and is expressed as a heparan, chondroitin, or keratan sulfate proteoglycan. Preconfluent basal keratinocytes, but not confluent or differentiated keratinocytes, also express chondroitin sulfate proteoglycan forms of CD44E and of the 180-kDa core protein. The modal size of the epican expressed on differentiated keratinocytes is smaller than the size of the epican expressed on basal keratinocytes. Thus, cell confluence and differentiation regulate several aspects of CD44 expression on keratinocytes, suggesting nuances in function for the different protein forms. PMID- 10478805 TI - Effects of exercise training on the heart rate variability and QT dispersion of patients with acute myocardial infarction. AB - Heart rate variability (HRV) reflects the autonomic tone of the heart, and QT dispersion reflects the regional inhomogeneity of ventricular repolarization. The purpose of the present study was to determine the effects of early exercise training on HRV and QT dispersion in patients with acute myocardial infarction (AMI). Forty patients (mean age: 59 years) with AMI were randomized to training rehabilitation (group Tr, n=20) or conventional rehabilitation (group C, n=20). Two weeks after AMI, group Tr underwent 10 min of exercise using a bicycle ergometer (80% of anaerobic threshold) twice a day. At the end of the second and fourth weeks, 12-lead and 24-h Holter ECGs were recorded. QT intervals were measured and corrected using Bazett's formula (QTc), and QTc dispersion (QTcd) was defined as the difference between maximum and minimum QTc. HRV was accessed by the high-frequency component (HF: 0.15-0.40 Hz) of the HRV power spectrum (parasympathetic activity) and the ratio of low frequency (0.04-0.15 Hz) to HF (L/H ratio: sympathetic activity). In group Tr, HF increased (82.5 to 131.1 ms2), the L/H ratio decreased (3.9 to 2.6), and QTcd decreased (77.2 to 57.2 ms). In group C, none of the indices changed. It was concluded that early exercise training improves sympathovagal balance and decreases QTcd, and may reduce the arrhythmogenic substrate following AMI. PMID- 10478806 TI - Prognostic value of serum hepatocyte growth factor in patients with acute coronary syndromes. AB - The present study examined whether or not hepatocyte growth factor (HGF), an endothelium-specific growth factor that stimulates regeneration of the endothelium, is increased or has a prognostic significance in patients with acute coronary syndromes. HGF was measured in 106 patients with coronary artery disease (20 stable effort angina, 12 unstable angina without adverse events, 24 unstable angina with adverse events and 50 acute myocardial infarction) on admission and 21 normal volunteers. The measurements in all patients were recorded before administration of heparin, and in acute myocardial infarction patients they were recorded from days 2 to 6 after heparin discontinuation on day 1. HGF levels (ng/ml) were 0.30+/-0.06 for the controls, 0.31+/-0.08 for stable effort angina patients, 0.31+/-0.08 for unstable angina patients without adverse events, 0.40+/ 0.20 for unstable angina patients with adverse events and in acute myocardial infarction patients they were 0.45+/-0.18 on day 0, 0.57+/-0.45 on day 2, 0.50+/ 0.35 on day 3, 0.48+/-0.32 on day 4, 0.44+/-0.20 on day 5, and 0.38+/-0.14 on day 6. HGF plays a crucial role in the restoration of injured endothelial cells and is a predictor of adverse events in patients with acute coronary syndromes. PMID- 10478807 TI - Effect of insulin resistance on the endothelial vasomotor function of the coronary artery of nondiabetic patients. AB - Attention has been paid to the relationship between insulin resistance and coronary artery disease. The present study investigated the relationship between insulin resistance and the endothelial vasomotor function of the coronary artery of nondiabetic patients with chest pain and a positive exercise tolerance test. Twenty-five nondiabetic patients with chest pain were included. Patients with a steady state plasma glucose (SSPG) level of greater than or equal to 135 mg/dl were placed in the insulin resistant (IR) group, and those with a SSPG level less than 135 mg/dl were placed in the noninsulin resistant (NIR) group. The effect of acetylcholine, papaverine, and isosorbide dinitrate on the vasomotor response of the coronary endothelium was studied. The percent change in diameter of the coronary artery after injection of acetylcholine (20 microg ml(-1) min(-1)) was 84+/-17% in the IR group, and 109+/-18% in the NIR group. The difference in the degree of the vasodilative response is statistically significant (p<0.01). The percent change in coronary flow velocity after injection of acetylcholine (20 microg ml(-1) min(-1)) in the IR group was 120+/-67%, whereas that in the NIR group was 256+/-58%; the increase in coronary artery flow velocity of the IR group was significantly smaller than that of the NIR group (p<0.01). Nondiabetic patients with insulin resistance have endothelial vasomotor dysfunction, which raises an important question as to whether nondiabetic patients with insulin resistance should be treated to prevent the development of coronary heart disease. PMID- 10478808 TI - Stretching of the ductus: an important factor in determining the outcome of coil occlusion. AB - The present study measured the minimal diameter of the ductus (minimal D), the stretched minimal diameter (stretched D), and the stretch index (SI) before coil occlusion in 25 patients with a patent arterial duct. The following factors were compared in the success group (22 cases, coil successfully placed after initial deployment) versus the failure group (3 cases): minimal D, stretched D, SI, the sum of the loop diameter of coils (the loop diameter), the sum of the product of the loop diameter and the number of loops (the loop diameter and number), the loop diameter/minimal D, the loop diameter/stretched D, the loop diameter and number/minimal D, and the loop diameter and number/stretched D. In the failure group, minimal D, stretched D, SI, the loop diameter, and the loop diameter and number were larger than in the success group. The loop diameter/stretched D, and the loop diameter and number/stretched D were smaller in the failure group. Although the loop diameter and number/minimal D was slightly smaller in the failure group, the loop diameter/minimal D was comparable. It is concluded that the stretched D is more reliable than minimal D to determine the appropriate size of coil for successful initial deployment. PMID- 10478809 TI - Long-term clinical and echocardiographic outcome in patients with mitral stenosis treated with percutaneous transvenous mitral commissurotomy. AB - Long-term follow-up after percutaneous transvenous mitral commissurotomy (PTMC) is limited. Ninety-four middle-aged (51+/-9 years) mitral stenosis patients who underwent successful PTMC were followed up with annual echocardiography for 6.1+/ 1.4 years. PTMC success was defined as either mitral valve area (MVA) >1.5 cm2 or a MVA of more than twice the pre-procedural value, together with no worsening of mitral regurgitation >grade 2+. Mitral valve replacement (MVR), worsening of congestive heart failure (CHF), and thromboembolism were sought for survival analysis. Restenosis was defined as loss of more than 50% of the initial procedural MVA gain. Functional limit of daily activities was assessed through a questionnaire. The study population was divided into group 1 (post-procedural MVA >2.0 cm2), group 2 (MVA > 1.5 cm2 and < or = 2.0 cm2) and group 3 (MVA < or = 1.5 cm2). The 6-year survival with freedom from MVR, CHF, thromboembolism, and combined events (MVR+CHF) was 92%, 95%, 91%, and 88%, respectively. No group 1 patient experienced MVR or CHF. Restenosis was predominant in group 3. Deterioration of daily activities during follow-up was not observed in group 1; however, it was significant in group 2 (p<0.05) and group 3 (p<0.001). These results demonstrated that patients who attained a large MVA (>2.0cm2) immediately after PTMC maintained their procedural benefit with less clinical complication and with less limitation of daily activity. PMID- 10478810 TI - Effects of single administration of a phosphodiesterase III inhibitor during cardiopulmonary bypass: comparison of milrinone and amrinone. AB - The effects of phosphodiesterase III (PDE III) inhibitors administered after aortic declamping during cardiopulmonary bypass (CPB) for open heart surgery were investigated. Ten patients (group M) were administered milrinone (50 microg/kg) after aortic declamping during CPB, 10 patients were administered amrinone (1 mg/kg) at the same time during their surgery (group A), and 10 patients served as controls with no drug administered (group C). Soon after bolus infusion of the PDE III inhibitor, perfusion pressure dropped significantly in groups M and A. However, after release of CPB and at the end of surgery, there was no difference in aortic pressure between the 3 groups. There were also no differences between the groups in heart rate, pulmonary artery pressure, and pulmonary capillary wedge pressure. After weaning from CPB, the cardiac index was high and systemic vascular resistance index was low in groups M and A. There were no significant differences in the need for additional catecholamines and time for rewarming between groups. No adverse reactions were observed. A single administration of a PDE III inhibitor during CPB was useful for post-CPB management of patients undergoing open heart surgery. Amrinone reduced perfusion pressures more than milrinone, but cardiac indices and aortic pressures after weaning from CPB showed no differences between group M and group A patients. PMID- 10478811 TI - Nonischemic ST-segment elevation induced by negative inotropic agents. AB - The present study investigated whether regional ventricular dyskinesia (ie, systolic bulging) is a direct cause of ST-segment elevation in canine hearts in vivo. Regional ventricular dyskinesia was induced in 33 anesthetized open-chest dogs by injection of negative inotropic agents into the left anterior descending coronary artery (LAD) without disruption of coronary blood flow. Regional myocardial contraction was assessed in terms of the percent systolic shortening (%SS) and percent systolic bulging (%bulging), which were measured using ultrasonic crystals. The ST-segment elevation of the LAD-perfused area was measured with a unipolar electrode. Lidocaine, a sodium channel blocker, nicorandil, a potassium channel opener, propranolol, a beta-adrenergic blocker, or verapamil, a calcium channel blocker, was administered by intracoronary injection during maximal vasodilation induced by adenosine. All drugs induced dose-dependent ST-segment elevation in association with a parallel reduction in %SS and a parallel increase in %bulging. The absence of myocardial ischemia was confirmed by the absence of NADH fluorescence. It was concluded that regional ventricular dyskinesia had an important role in ST-segment elevation not associated with myocardial ischemia. PMID- 10478812 TI - Effects of low doses of endothelin-1 on basal vascular tone and autoregulatory vasodilation in canine coronary microcirculation in vivo. AB - The plasma level of endothelin-1 (ET-1) increases in several cardiovascular disorders. The present study examined whether threshold doses of ET-1 affect vascular tone and autoregulatory vasodilation during a reduction in perfusion pressure in the coronary microcirculation in vivo. In anesthetized open-chest dogs, arterial microvessels in the epimyocardium were observed through a microscope equipped with a floating objective. In 6 dogs, ET-1 (10(-13) to 10( 8)mol/L) was superfused onto the epimyocardium in a cumulative fashion. In another set of dogs (n= 16), the perfusion pressure of the observed vascular bed was reduced to 60 mmHg (mild stenosis) and to 40 mmHg (severe stenosis) by a hydraulic occluder, and the microvascular responses were observed in the presence (n=9) or absence (n=7) of ET-1 (10(-12) or 10(-11) mol/L). ET-1 > or =10(-11) mol/L constricted coronary arterioles (< or =100 microm in diameter) and small arteries (>100 microm in diameter) in a dose-dependent fashion. ET-1 of 10(-12) mol/L affected neither the basal diameters nor the dilation of vessels during the pressure reduction. ET-1 of 10(-11) mol/L decreased the diameters of arterioles and small arteries before and during the mild and severe stenosis. However, ET-1 did not attenuate the percentage dilation of arterioles from the baseline in response to the mild and severe stenosis. The data indicates the following: (1) ET-1 at doses > or =10(-11) mol/L similarly constricts coronary arterioles and small arteries; (2) ET-1 at 10(-11) mol/L, which is slightly higher than the pathophysiological plasma level, increases the basal vascular tone, but does not attenuate the autoregulatory vasodilation of the coronary microcirculation. PMID- 10478813 TI - Stretch-activated ion channel blocker gadolinium attenuates ischemic ST-segment elevation in canine myocardium. AB - Regional transmural ischemia causes both ST-segment elevation and systolic elongation (bulging) of the myocardium. Mechanical stretch might alter the transmembrane potential via stretch-activated ion channels (SAC); however, the role of SAC on ischemic ST-segment elevation has not yet fully studied. The present study investigated the role of SAC in the genesis of ischemic ST-segment elevation in the in-vivo canine heart. In 6 anesthetized dogs, an extracorporeal conduit connected to the left anterior descending coronary artery was occluded for 5 min before and after the intracoronary infusion of gadolinium (Gd, 500 micromol/min). To eliminate the effect of ischemic preconditioning, the bypass was occluded for 5 min before the experiment. Percent systolic shortening (%SS) and percent systolic elongation (%bulging) were measured using a pair of ultrasonic dimension crystals. A unipolar epicardial ECG was monitored at the center of the ischemic area for the measurement of the ST-segment level. At the end of coronary occlusion, there was no difference in the reduction of %SS or the increase of %bulging between before and after infusion of Gd. ST-segment elevation, however, was significantly attenuated by the infusion of Gd. These data demonstrated that the activation of SAC is one cause of ischemic ST-segment elevation. PMID- 10478814 TI - Heart rate variability analysis of patients with idiopathic left ventricular outflow tract tachycardia: role of triggered activity. AB - There have been several reports with respect to idiopathic ventricular tachycardias (VTs) originating from the left ventricular outflow tract (LVOT). A previous report suggested that triggered activity plays a partial role in idiopathic LVOT tachycardia from the electrophysiological as well as the electropharmacological viewpoint. However, the exact role of triggered activity in this type of VT remains unknown. In the present study the relationship of the frequency of premature ventricular contractions (PVCs) and heart rate was examined and heart rate variability (HRV) was analyzed in 2 cases of LVOT tachycardia using 24-h Holter electrocardiographic (ECG) monitoring. The relation between the PVCs frequency and heart rate showed a persistently positive correlation, indicating frequent PVCs as heart rate increased. In HRV analysis, NN50(%), a time-domain variable of parasympathetic activity, showed no change prior to ventricular arrhythmias. In frequency-domain analysis of HRV, the high frequency (HF) component tended to fall prior to repetitive PVCs and VTs. The ratio of the low frequency to high frequency (LF/HF) components increased prior to single PVCs, repetitive PVCs and VTs. Sympathetic predominance predisposes the genesis of these kinds of arrhythmias originating from the LVOT and it is suggested that triggered activity plays an important role in LVOT tachycardia, at least in its initiation. PMID- 10478815 TI - Angioplasty-bypass surgery combination therapy: case study of an elderly patient with unstable angina and colon cancer. AB - An 83-year-old man was admitted with refractory unstable angina and severe anemia. Colonofiberscopy revealed hemorrhagic colon cancer in the transverse colon. Coronary angiography showed total occlusion of the right coronary artery (RCA), diffuse, calcified 90% stenosis of the middle portion of the left anterior descending artery (LAD); and fair collaterals from the LAD to the RCA. Coronary revascularization was considered prior to colectomy, but because of the patient's advanced age and hemorrhagic cancer, conventional coronary aorta bypass grafting (CABG) using extracorporeal circulation, as well as coronary stenting requiring antiplatelet therapy, were regarded as inadvisable. Percutaneous transluminal coronary angioplasty (PTCA) for the LAD carried the risk of suboptimal coronary stenting. Thus, the patient was first treated with PTCA for the occluded RCA, followed 7 days later by a left internal thoracic artery graft to the LAD on the beating heart without extracorporeal circulation. The patient was stable thereafter. This approach to coronary revascularization may be suitable for patients for whom anticoagulation or antiplatelet therapy are contraindicated, or when complete revascularization would be difficult with CABG or PTCA alone. PMID- 10478816 TI - Dynamic outflow obstruction due to the transient extensive left ventricular wall motion abnormalities caused by acute myocarditis in a patient with hypertrophic cardiomyopathy: reduction in ventricular afterload by disopyramide. AB - A 65-year-old woman was admitted to the coronary care unit because of acute pulmonary edema. Immediate 2-dimensional and Doppler echocardiograms revealed extensive left ventricular wall motion abnormalities and left ventricular hypertrophy with extreme outflow obstruction. Although an ECG showed ST-segment elevation in the anterolateral leads, a coronary arteriogram revealed normal epicardial arteries. Heart failure was relieved after diminishing the dynamic outflow obstruction with disopyramide administration. An endomyocardial biopsy from the right ventricle on the 8th hospital day showed borderline myocarditis. Wall motion abnormalities gradually normalized within 2 weeks. It is speculated that her pulmonary edema would not have been relieved so readily without the immediate reduction in ventricular afterload by disopyramide. These clinical changes over time were observed with serial echo-Doppler examinations. PMID- 10478817 TI - Myocardial infarction with Moyamoya disease and pituitary gigantism in a young female patient. AB - Myocardial infarction is very rare in young female patients with systemic vascular disorders. Moyamoya disease is a cerebrovascular disease associated with an abnormal vascular network. This report presents a 19-year-old female patient who suffered from chest pain and exertional dyspnea for 2 months prior to admission. She had a history of Moyamoya disease and pituitary gigantism since childhood. Her ejection fraction on echocardiogram was 20% and a perfusion defect with partial reversibility in the anterior wall was demonstrated on stress single photon emission computed tomography (SPECT). Diagnostic coronary angiogram revealed critical stenosis in the middle left anterior descending artery, which was treated by coronary stenting. Her subjective symptoms were relieved and the perfusion defect seen on SPECT decreased after coronary intervention. PMID- 10478818 TI - Exercise-induced uncommon atrioventricular nodal reentrant tachycardia with sick sinus syndrome: a case report. AB - Exercise seldom provokes tachycardia in patients with paroxysmal supraventricular tachycardia (PSVT). This report presents a case of exercise-induced uncommon atrioventricular nodal reentrant tachycardia (AVNRT) with sick sinus syndrome. Treadmill exercise testing provoked AVNRT of long RP' with good reproducibility. Uncommon AVNRT was confirmed by the lack of atrial pre-excitation during PSVT and para-Hisian pacing. The patient has been successfully treated with verapamil and DDD pacing for 5 years. PMID- 10478819 TI - Thrombosed dissection of the ascending aorta complicating extravasation. AB - This report presents 2 patients with thrombosed dissection of the ascending aorta complicating extravasation. The first case was an 85-year-old male admitted with shock due to cardiac tamponade. Plain computed tomography (CT) demonstrated a dilated ascending aorta without clear evidence of aortic dissection. The second case was a 77-year-old female presenting with shock, in whom an enhanced CT scan demonstrated a dilated ascending aorta and periaortic effusion. However, dissection of the distal ascending aorta was not identified in either case before emergency surgery. In case 1, soon after the bloody pericardial effusion was decompressed during the operation, bleeding from the ascending aorta occurred. A small intimal tear was found in the distal ascending aorta, and in each case the pseudolumen was filled with fresh thrombus. The ascending aorta was replaced. Each patient had an uneventful postoperative recovery. Based on this experience, it is suggested that patients with thrombosed ascending aortic dissection complicating extravasation should undergo early graft replacement. PMID- 10478820 TI - Pharmacokinetics of TCDD in veterans of Operation Ranch Hand: 15-year follow-up. AB - Using multiple measurements from serum collected over 15 yr (in 1982, 1987, 1992, and 1997), the half-life of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in 97 veterans of Operation Ranch Hand, the Air Force unit responsible for the aerial spraying of Agent Orange in Vietnam, was estimated. The potential influences of age, percent body fat, and changes in percent body fat on the half-life estimate were also examined. The elimination rate of TCDD for these veterans was 0.0915 per year with 95% confidence interval 0.0844 to 0.0986 per year; the corresponding half-life estimate was 7.6 yr with 95% confidence interval 7.0 to 8.2 yr. The elimination rate decreased significantly with increasing body fat, but not with age or relative changes in percent body fat. PMID- 10478821 TI - Toxicity and bioaccumulation of nickel sulfate in Sprague-Dawley rats following 13 weeks of subchronic exposure. AB - Adult male Sprague-Dawley rats were given 0, 0.02, 0.05, and 0.1% nickel sulfate (NiSO4-6H2O) or 0, 44.7, 111.75, and 223.5 mg Ni/L, respectively, in their drinking water for 13 wk. Twenty-four hours following the end of such treatment, all animals survived and no apparent clinical signs of toxicity were noted. The final mean body weights of various nickel sulfate-treated rats were not significantly decreased except for the 0.1% nickel sulfate treated group when compared to those in the control. The absolute and relative organ weights were either increased or decreased or remained unchanged, depending on the organ and the dose of nickel sulfate. Total plasma proteins, plasma albumin and globulins, and plasma glutamic pyruvic transaminase activity were all significantly decreased in 0.1% nickel sulfate-treated rats. Lymphocyte subpopulations (T and B cells) were induced at lower dose levels, but suppressed at the highest (0.1%) dose group. A significant decrease in urine volume and an increase in BUN were observed at the highest dose group. Biochemical analysis of bronchoalveolar lavage fluid and lung tissue showed some lung damage, whereas no damage to the testis or DNA in liver and kidneys were found. No gross or microscopic changes were seen in any of the various tissues examined. The relative order of bioaccumulation of nickel in different organs of rats when treated at 0.1% nickel sulfate (223.5 mg Ni/L) was kidneys > testes > lung = brain > spleen > heart = liver. But with regard to order of toxicity, both immune and pulmonary systems were found to be very sensitive targets, followed by kidney. PMID- 10478822 TI - Cadmium-induced renal lipid peroxidation in rats and protection by selenium. AB - Cadmium has been recognized as one of the most toxic environmental and industrial pollutants. The kidney is a critical target organ following Cd exposure. The aim of this study was to investigate the effects of cadmium-induced peroxidative damage to rat kidney. Treatment of rats with Cd resulted in a time- and dose related accumulation of metal in kidney. Cd produced enhanced lipid peroxidation in plasma and kidney. These Cd-induced changes were accompanied by a significant rise in renal Fe and Cu, and a fall in tissue Zn and Se. Concurrent treatment with Se and Cd reduced the Cd-induced alterations in renal peroxidation and essential metal levels. Data suggest that lipid peroxidation is associated with Cd toxicity and that Se was found effective in attenuation of these renal effects. PMID- 10478823 TI - Liver mitochondria alterations in chloroform-treated Sprague-Dawley rats. AB - The formation of a chloroform adduct produced by the reaction of the oxidative chloroform metabolite phosgene with two molecules of phosphatidylethanolamine has been previously demonstrated in liver mitochondria of phenobarbital-pretreated Sprague-Dawley (SD) rats. The aim of our study was to assess the influence of chloroform adduct mitochondrial accumulation on the hepatic mitochondria morphology. Liver mitochondrial ultrastructural alterations were analyzed by electron microscopy in SD rats administered with increasing doses of chloroform. Variation in the morphology of mitochondria, consisting of an increase of intertwined organelles, only rarely seen in control specimens, was observed at the lowest chloroform dose (180 mg/kg). At higher doses, mitochondrial damage progressed with swelling of the organelles and formation of megamitochondria. These megamitochondria were characterized by a dilution of the matrix, and often membranous whorls were found inside the matrix. The two distinct forms of cell death, necrosis and apoptosis, were first observed at 300 mg/kg of chloroform. Our results suggest that the formation and the accumulation of a chloroform modified phosphatidylethanolamine in mitochondria induce ultrastructural modifications of these organelles. In conclusion, mitochondria are involved in chloroform-induced hepatotoxicity. PMID- 10478824 TI - Activity of cathepsin G, elastase, and their inhibitors in plasma during methanol intoxication. AB - Methanol oxidation in the liver is accompanied by formation of formaldehyde and free radicals. These compounds can react with biologically active proteins, including proteolytic enzymes and their inhibitors. The activity of cathepsin G and elastase and their inhibitors such as alpha-1-antitrypsin and alpha-2 macroglobulin in plasma of rats given methanol orally in doses of 1.5, 3, and 6 g/kg was investigated for 7 days. The activity of cathepsin G and elastase was increased from 12 h to 5 d, proportionally to methanol dose. At the same time, activity of their inhibitors was reduced. Methanol ingestion in humans caused changes in activities of proteases and their inhibitors with similar direction as in rats. These changes in activity of proteases and their inhibitors produce significant disturbances in proteolytic-antiproteolytic balance after methanol administration. PMID- 10478825 TI - Inhibition of eleven mutagens by various tea extracts, (-)epigallocatechin-3 gallate, gallic acid and caffeine. AB - The antimutagenic properties of various tea extracts (green tea, pauchong tea, oolong tea and black tea) and their components including (-)-epigallocatechin-3 gallate (EGCG), gallic acid and caffeine were examined by the Ames test. The antimutagenic activity of the green tea extract against N-methyl-N'-nitro-N nitrosoguanidine (MNNG), folpet and monocrotophos was greater than those of pouchong, oolong and black tea extracts. The antimutagenic effects of tea extracts against 2-acetylaminofluorene (AAF) decreased as follows: oolong tea > pauchong tea > black tea > green tea. Furthermore, black tea showed a greater antimutagenic activity against benzo[a]pyrene (BP). The pauchong tea showed a stronger inhibitory effect against 9-aminoacridine (9AA) and aflatoxin B1 (AFB1) than other tea extracts. EGCG markedly suppressed the direct-acting mutagenicity of MNNG, N-nitroso-N-methylurea (MNU), captan, and folpet which were alkylating agents and fungicides. Similarly, gallic acid, the major component of black tea strongly inhibited the mutagenicity of 9AA, and moderately inhibited the mutagenicity of MNNG and folpet. The caffeine was less active. EGCG and gallic acid perhaps could act as nucleophiles to scavenge the electrophilic mutagens. Taken together, these results suggest that formation of different metabolites during various stages of tea fermentation may affect antimutagenic potencies against different types of chemical mutagens. PMID- 10478826 TI - Co-mutagenicity of coumarin (1,2-benzopyrone) with aflatoxin B1 and human liver S9 in mammalian cells. AB - Coumarin (1,2-benzopyrone), a natural dietary constituent and drug currently under evaluation for treatment of certain cancers and lymphedema, reduces polycyclic aromatic hydrocarbon-induced neoplasms in rodents. Because most rodents metabolize coumarin through 3,4-epoxidation, whereas 7-hydroxylation predominates in humans, their suitability as a model for coumarin effects in humans has been questioned. We examined coumarin chemoprotection against the promutagen and dietary contaminant aflatoxin B1 with human liver S9 bioactivation in the Chinese hamster ovary cell/hypoxanthine-guanine phosphoribosyltransferase mutation assay. Coumarin in the absence of aflatoxin B1 was not mutagenic or cytotoxic up to 500 microM. When included with either 1 or 10 microM aflatoxin B1, coumarin produced a dose-dependent increase in mutant frequency and cytotoxicity. At concentrations greater than 50 microM, coumarin stimulated human liver S9 bioactivation of aflatoxin B1 to the mutagenic 8,9-epoxide. This increase was 12- and fivefold at 500 microM coumarin with 1 and 10 microM aflatoxin B1, respectively, compared with incubations with aflatoxin B1 alone. These findings differ from previous results with liver S9 from other species, and indicate that coumarin co-mutagenicity with aflatoxin B1 and human liver S9 is through increased aflatoxin B1 bioactivation. PMID- 10478828 TI - Potentiation of paracetamol and carbon tetrachloride-induced hepatotoxicity in rodents by the food additive vanillin. AB - The potential of vanillin to potentiate the paracetamol and carbon tetrachloride (CCl4)-induced hepatotoxicity was investigated in rats. Vanillin when given alone (15 mg/kg, orally), did not modify liver function in rats as the values of serum enzymes of alkaline phosphatase (ALP) and aminotransaminases (AST and ALT) were found similar to those in the normal animals. However, when given repeatedly before the administration of the subtoxic dose of paracetamol (500 mg/kg) or CCl4 (1 ml/kg), vanillin caused liver damage, as manifested by the significant increase in the serum levels of hepatic enzymes. When tested for its possible interaction with pentobarbital (75 mg/kg, i.p.) and strychnine (0.9 mg/kg, i.p.), it caused reduction in pentobarbital-induced sleep in mice as well as preventing the animals against the lethal effect of strychnine, suggestive of an induction of microsomal drug metabolizing enzymes. These results indicate that vanillin potentiates the hepatotoxic potential of paracetamol and CCl4 in rats probably through an enzyme induction process. PMID- 10478827 TI - Effects of roselle (Hibiscus sabdariffa Linn.), a Thai medicinal plant, on the mutagenicity of various known mutagens in Salmonella typhimurium and on formation of aberrant crypt foci induced by the colon carcinogens azoxymethane and 2-amino 1-methyl-6-phenylimidazo[4,5-b]pyridine in F344 rats. AB - The 80% ethanol extract of roselle (Hibiscus sabdariffa Linn.), a medicinal plant in Thailand, was examined for antimutagenic and chemopreventive activity in a colon carcinogenesis model. It reduced about 60-90% of the mutagenicity induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and other heterocyclic amines 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,4 dimethylimidazo[4,5-f]quinoline(MelQ),2-amino-3, 8-dimethylimidazo[4,5 f]quinoxaline(MelQx),3-amino-1,4-dimet hyl-5H-pyrido[4,3-b]indole (Trp-P-1), 3 amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2),2-amino-6-methyldipyrido[1,2 a:3',2'-d]imidazole (Glu-P-1),2-aminodipyrido[1,2-a:3',2'-d]imidazole (Glu-P-2), at a concentration of 12.5 mg/plate in the Salmonella mutation assay. The extract showed no mutagenicity and no antibacterial activity below this dose. Mutagenicity of methylazoxymethanol (MAM) acetate, which, like PhIP, is a colon carcinogen,was also efficiently inhibited by the roselle extract. To investigate chemoprevention by roselle in a colon carcinogenesis model, we examined the inhibitory effects of the roselle extract in F344 rats in which aberrant crypt focus (ACF) formation was induced by azoxymethane (AOM) and PhIP. In the initiation stage, the number of AOM-induced ACF in the colon was significantly decreased by roselle (17-25%) compared with that in rats treated with AOM alone. The amount of O6-methylguanine in the colonic mucosa tended to be decreased in the roselle-treated rats. The number of PhIP-induced ACF was also significantly decreased by roselle treatment (22%) at a concentration of 1.0 g/kg body weight in the initiation stage. However, in the post-initiation stage of AOM-induced ACF formation, roselle increased the number of ACF, especially the number of foci which had more than three crypts/focus. These results indicate that roselle has antimutagenic activity against MAM acetate and heterocyclic amines and that it decreases the number of AOM- and PhIP-induced ACF in the initiation stage, although it rather increased the number of ACF in the post-initiation stage. PMID- 10478829 TI - Effect of some indole derivatives on xenobiotic metabolism and xenobiotic-induced toxicity in cultured rat liver slices. AB - In this study the effect of some indole derivatives on xenobiotic metabolizing enzymes and xenobiotic-induced toxicity has been examined in cultured precision cut liver slices from male Sprague-Dawley rats. While treatment of rat liver slices for 72 hours with 2-200 microM of either indole-3-carbinol (I3C) or indole 3-acetonitrile (3-ICN) had little effect on cytochrome P-450 (CYP)-dependent enzyme activities, enzyme induction was observed after in vivo administration of I3C. The treatment of rat liver slices with 50 microM 3,3'-diindolylmethane (DIM; a dimer derived from I3C under acidic conditions) for 72 hours resulted in a marked induction of CYP-dependent enzyme activities. DIM appears to be a mixed inducer of CYP in rat liver slices having effects on CYP1A, CYP2B and CYP3A subfamily isoforms. Small increases in liver slice reduced glutathione levels and glutathione S-transferase activity were also observed after DIM treatment. While aflatoxin B1 and monocrotaline produced a concentration-dependent inhibition of protein synthesis in 72-hour-cultured rat liver slices, cytotoxicity was markedly reduced in liver slices cultured with 50 microM DIM. These results demonstrate that cultured rat liver slices may be employed to evaluate the effects of chemicals derived from cruciferous and other vegetables on CYP isoforms. In addition, liver slices can also be utilized to examine the ability of such chemicals to modulate xenobiotic-induced toxicity. PMID- 10478830 TI - Comparative DNA cross-linking by activated pyrrolizidine alkaloids. AB - The toxicity and bioactivity of pyrrolizidine alkaloids (PAs), common constituents of hundreds of plant species, and in herbal remedies and folk medicines prepared thereof, are probably due to their ability to form DNA cross linking. We investigated DNA cross-linking activity by chemically-activated PAs from four different structural classes in Madin-Darby bovine kidney (MDBK) cells and in pBR322 DNA. In cell culture, alpha,beta-unsaturated macrocyclic diester pyrroles dehydrosenecionine (DHSN), dehydroriddelliine (DHRD) and the saturated macrocyclic diester pyrrole dehydromonocrotaline (DHMO) were significantly more potent cross-linkers than the simple necine base (retronecine) and an N-oxide (indicine N-oxide; INO) as determined by alkaline elution. The proportion of total DNA cross-links that were proteinase K-resistant (DNA-DNA cross-links) induced by the various pyrroles ranged from 0.08 (DHRN) to 0.67 (DHSN). Those pyrroles that were potent cross-linkers of cellular DNA also cross-linked, in a dose-dependent manner, Bam HI-digested pBR322 DNA as assessed by a gel retardation assay. The possible functional relevance of pyrrole-DNA cross-links was determined by their ability to interrupt PCR amplification of a 1.129 kb segment of pBR322. Dehydrosenecionine completely inhibited amplification, while DHMO was of intermediate potency, while DHRN and INO had no effect. Taken together, these studies suggest that structural features, most notably the presence of a macrocyclic diester, confer potent cross-link activity to PAs. In any event, DNA-DNA cross-linking is probably biologically relevant as indicated by their interference with DNA replication. PMID- 10478831 TI - Enhancement of ovalbumin-induced antibody production and mucosal mast cell response by mercury. AB - Food contaminants may contribute to the recent increased incidence of food allergies. We have investigated this hypothesis experimentally. It was our objective to determine whether toxicity to the intestinal tissue by orally applied mercury (Hg) could modulate the immune response to food allergens. Effective mechanisms were studied with functional immunological and toxicological parameters. Brown Norway rats were immunized intraperitoneally by ovalbumin (OVA). Before oral challenge with OVA, immunized and non-immunized animals were exposed to HgCl2. Immunological responses were measured by enzyme-linked immunosorbent assays [anti-OVA-IgE and-IgG, rat mast cell protease II (RMCPII), interferon-gamma, interleukin-4, lymphocyte proliferation] and by flow cytometry (lymphocyte subpopulations). Toxicity of Hg to the intestinal barrier was determined by measuring viability, DNA damage and induction of glutathione S transferase in isolated intestinal epithelial cells and lymph node cells, and by measuring permeability, short-circuit current and tissue conductance of the intact intestinal epithelium. A single high oral dose of HgCl2 enhanced the serum concentrations of anti-OVA-IgE and IgG (P < 0.05) and of RMCPII (P < 0.05) in immunized rats. The treatment resulted in a higher number of CD4/CD25+ T cells in the lymph nodes (P < 0.05). The multiple application of low HgCl2 doses (5 x 0.2 mg/kg body weight) only resulted in an elevated RMCPII serum concentration (P < 0.05). Neither treatment schedules impaired proliferation and cytokine production of lymphocytes. In non-immunized rats only minor immunological changes were observed. Oral HgCl2 induced genotoxic damage in lymph node cells and in jejunal epithelial cells (P < 0.05). Moreover, HgCl2 increased the permeability of intestinal epithelial tissue and of Caco-2 monolayers and was genotoxic and cytotoxic to isolated intestinal epithelial cells in vitro. In conclusion, these studies indicate that the food contaminant Hg can stimulate the immune response to OVA in immunized rats. One possible mechanism could be the toxicity of Hg to the intestinal epithelial and the lymph node cells. Whether humans with allergies respond to high oral doses of Hg in a similar way needs to be investigated in further studies. PMID- 10478832 TI - In vitro exposure of human lymphocytes to trichothecenes: individual variation in sensitivity and effects of combined exposure on lymphocyte function. AB - The trichothecenes are mycotoxins produced by fungi of the genus Fusarium, which are commonly present in foods and feed of cereal origin. Owing to the lack of sufficient toxicological data for most of the trichothecenes, in vitro studies may contribute to risk assessments of these toxins. In the present report, human lymphocyte cultures were used to study the individual variation in sensitivity among humans and the effects on in vitro Ig production. Furthermore, proliferative responses of cells exposed to combinations of two of the toxins were studied. Four toxins, T-2 toxin, diacetoxyscirpenol (DAS), nivalenol (NIV) and deoxynivalenol (DON) were included in the study. All four of the tested trichothecenes effectively inhibited mitogen-induced lymphocyte proliferation. There were no statistically significant differences in sensitivity to the toxins between lymphocytes from female and male blood donors. The individual variation in sensitivity, evaluated as the range of IC50 values, was rather limited (within a factor of 3 to 4). Immunoglobulin production by pokeweed-stimulated human lymphocytes was also effectively inhibited with IC50 values similar to the IC50 values in the proliferation tests for DON and NIV. However, IC50 values for Ig synthesis in cultures exposed to T2 were approximately two to three times higher than the corresponding IC50 values found in the proliferation tests. At low levels of exposure, elevated Ig production was observed in lymphocyte cultures from four out of the five blood donors tested. This effect was most pronounced on IgA synthesis. Combinations of NIV with T2, DAS or DON resulted in additive toxicity in the lymphocyte proliferation test, while combinations of DON with T2 or DAS resulted in an inhibition that was slightly lower than what could have been expected from the inhibition produced by the individual toxins. In conclusion, the tested trichothecenes inhibited both proliferation and Ig production in human lymphocytes in a dose-dependent manner with limited variation in sensitivity between individuals. Enhanced Ig production was observed in cell cultures exposed to the lower doses of the toxins. Combined exposure to two of the toxins resulted mainly in additive or antagonistic effects, although synergistic effects cannot be excluded and should be further investigated. These findings indicate that the total intake of type A and B trichothecenes should be taken into account in risk assessments. PMID- 10478833 TI - Assessment of the no-observed-effect level (NOEL) of quinalphos in pregnant rats. AB - Technical quinalphos (0.5, 1.5, 2, 3 or 4.5 mg/kg body weight) was administered orally to pregnant rats from day 6-15 of gestation. At 3 and 4.5 mg/kg/day, quinalphos produced significant changes in hepatic ALT, ALP and serum ALT, AST, ALP and LDH activity along with hepatocellular changes in dams. The AchE activity in brain and red blood cells was also significantly inhibited at these two doses. At 0.5, 1.5 and 2 mg/kg/day, however, quinalphos did not produce any such changes. Up to a dose of 2 mg/kg/day there was no foetotoxic or teratogenic effect, as evidenced by number of implantation sites, percent resorption, foetal weight, morphological, visceral and skeletal evaluations. Hence, 2 mg/kg body weight of quinalphos could be considered as the no-observed-effect level (NOEL) on foetal and maternal toxicity in rats. PMID- 10478834 TI - The effects of vitamin E on arylamine N-acetyltransferase activity in strains of Helicobacter pylori from peptic ulcer patients. AB - Arylamine N-acetyltransferase (NAT) activities with 2-aminofluorene (2-AF) and p aminobenzoic acid (PABA) were determined in the bacterium Helicobacter pylori. Cytosols or suspensions of H. pylori, with or without specific concentrations of vitamin E co-treatment, showed different percentages of 2-AF acetylation. The data indicated that there was increased NAT activity associated with increased levels of vitamin E in H. pylori cytosols and intact bacteria. For the cytosol and intact bacteria examinations, the apparent values of Km and Vmax were increased when vitamin E was added to the reaction mixtures for 2-AF and PABA acetylation, respectively. This report is the first demonstration to show that antioxidant agents (vitamin E) can promote H. pylori N-acetyltransferase activity. PMID- 10478835 TI - Modulatory potential of clocimum oil on mouse skin papillomagenesis and the xenobiotic detoxication system. AB - The present study was designed to elucidate the mechanistic inhibitory efficacy of clocimum (an eugenol rich variety of Ocimum gratissimum; Labiatae) oil on murine skin papillomagensis. Topical application of clocimum oil (50 microl/animal/day) during peri-initiation stage (1 week before and 2 weeks after initiation) of 7,12-dimethylbenz[a]anthracene (DMBA)-induced papillomagenesis and/or during the tumour promotion stage reduced (P < 0.05) the (i) tumour burden to 5.00, 4.41 and 4.50 (positive control value 5.27); (ii) cumulative number of papillomas to 85, 75 and 72 (positive control value : 95); and (iii) percent incidence of mice bearing papillomas to 94, 89 and 88, respectively (positive control value 94). Significant (P < 0.01) elevation in the hepatic levels of glutathione S-transferase (GST), sulfhydryl (-SH) and cytochrome b5 (Cyt. b5) was observed by the respective topical treatment of clocimum oil. Even in the skin tissue of the mouse, the topical treatment of clocimum oil enhanced (P < 0.05) the -SH content. The results suggest the modest chemopreventive potential of clocimum oil against the murine skin papillomagenesis, and such effects may be partly due to the modulated xenobiotic detoxication system enzymes. PMID- 10478837 TI - Chronic hypersecretion of luteinizing hormone in transgenic mice selectively alters responsiveness of the alpha-subunit gene to gonadotropin-releasing hormone and estrogens. AB - Steroid hormones can act either at the level of the hypothalamus or the pituitary to regulate gonadotropin subunit gene expression. However, their exact site of action remains controversial. Using the bovine gonadotropin alpha-subunit promoter linked to an expression cassette encoding the beta-subunit of LH, we have developed a transgenic mouse model where hypersecretion of LH occurs despite the presence of elevated ovarian steroids. We used this model to determine how hypersecretion of LH could occur when steroid levels are pathological. During transition from the neonatal period to adulthood, the endogenous LHbeta subunit gene becomes completely silent in these mice, whereas the alpha-directed transgene and endogenous alpha-subunit gene remain active. Interestingly, gonadectomy stimulates expression of the endogenous alpha and LHbeta subunit genes as well as the transgene; however, only the endogenous LHbeta gene retains responsiveness to 17beta-estradiol and GnRH. In contrast, LH levels remain responsive to negative regulation by androgen. Thus, alpha-subunit gene expression, as reflected by both the transgene and the endogenous gene, has become independent of GnRH regulation and, as a result, unresponsive to estradiol negative feedback. This process is accompanied by a decrease in estrogen receptor alpha gene expression as well as an increase in the expression of transcription factors known to regulate the alpha-subunit promoter, such as cJun and P-LIM. These studies provide in vivo evidence that estrogen-negative feedback on alpha and LHbeta subunit gene expression requires GnRH input, reflecting an indirect mechanism of action of the steroid. In contrast, androgen suppresses alpha subunit expression in both transgenic and nontransgenic mice. This suggests that androgens must regulate alpha-subunit promoter activity independently of GnRH. In addition to allowing the assessment of site of action of sex steroids on alpha subunit gene expression, these studies also indicate that chronic exposure of the pituitary to LH-dependent ovarian hyperstimulation leads to a heretofore undescribed pathological condition, whereby normal regulation of alpha, but not LHbeta, subunit gene expression becomes compromised. PMID- 10478836 TI - Interaction of radicicol with members of the heat shock protein 90 family of molecular chaperones. AB - The Hsp90 family of proteins in mammalian cells consists of Hsp90 alpha and beta, Grp94, and Trap-1 (Hsp75). Radicicol, an antifungal antibiotic that inhibits various signal transduction proteins such as v-src, ras, Raf-1, and mos, was found to bind to Hsp90, thus making it the prototype of a second class of Hsp90 inhibitors, distinct from the chemically unrelated benzoquinone ansamycins. We have used two novel methods to immobilize radicicol, allowing for detailed analyses of drug-protein interactions. Using these two approaches, we have studied binding of the drug to N-terminal Hsp90 point mutants expressed by in vitro translation. The results point to important drug contacts with amino acids inside the N-terminal ATP/ADP-binding pocket region and show subtle differences when compared with geldanamycin binding. Radicicol binds more strongly to Hsp90 than to Grp94, the Hsp90 homolog that resides in the endoplasmic reticulum. In contrast to Hsp90, binding of radicicol to Grp94 requires both the N-terminal ATP/ADP-binding domain as well as the adjacent negatively charged region. Radicicol also specifically binds to yeast Hsp90, Escherichia coli HtpG, and a newly described tumor necrosis factor receptor-interacting protein, Trap-1, with greater homology to bacterial HtpG than to Hsp90. Thus, the radicicol-binding site appears to be specific to and is conserved in all members of the Hsp90 family of molecular chaperones from bacteria to mammals, but is not present in other molecular chaperones with nucleotide-binding domains. PMID- 10478839 TI - Essential requirement for Pax6 in control of enteroendocrine proglucagon gene transcription. AB - The primary function of islet A cells is the synthesis and secretion of glucagon, an essential hormonal regulator of glucose homeostasis. The proglucagon gene is also expressed in enteroendocrine L cells of the intestinal epithelium, which produce glucagon-like peptide 1 (GLP-1) and glucagon-like peptide 2 (GLP-2), regulators of insulin secretion and intestinal growth, respectively. We show here that Pax6, a critical determinant of islet cell development and proglucagon gene expression in islet A cells, is also essential for glucagon gene transcription in the small and large intestine. Pax6 is expressed in enteroendocrine cells, binds to the G1 and G3 elements in the proglucagon promoter, and activates proglucagon gene transcription. The dominant negative Pax6 allele, SEYNeu, represses proglucagon gene transcription in enteroendocrine cells. Mice homozygous for the SEYNeu mutation exhibit markedly reduced levels of proglucagon mRNA transcripts in the small and large intestine, and GLP-1 or GLP-2-immunopositive enteroendocrine cells were not detected in the intestinal mucosa. These findings implicate an essential role for Pax6 in the development and function of glucagon producing cells in both pancreatic and intestinal endodermal lineages. PMID- 10478838 TI - Developmental and hormonal regulation of murine scavenger receptor, class B, type 1. AB - The scavenger receptor, class B, type I (SR-BI), is the predominant receptor that supplies plasma cholesterol to steroidogenic tissues in rodents. We showed previously that steroidogenic factor-1 (SF-1) binds a sequence in the human SR-BI promoter whose integrity is required for high-level SR-BI expression in cultured adrenocortical tumor cells. We now provide in vivo evidence that SF-1 regulates SR-BI. During mouse embryogenesis, SR-BI mRNA was initially expressed in the genital ridge of both sexes and persisted in the developing testes but not ovary. This sexually dimorphic expression profile of SR-BI expression in the gonads mirrors that of SF-1. No SR-BI mRNA was detected in the gonadal ridge of day 11.5 SF-1 knockout embryos. Both SR-BI and SF-1 mRNA were expressed in the cortical cells of the nascent adrenal glands. These studies directly support SF-1 participating in the regulation of SR-BI in vivo. We examined the effect of cAMP on SR-BI mRNA and protein in mouse adrenocortical (Y1-BS1) and testicular carcinoma Leydig (MA-10) cells. The time courses of induction were strikingly similar to those described for other cAMP- and SF-1-regulated genes. Addition of lipoproteins reduced SR-BI expression in Y1-BS1 cells, an effect that was reversed by administration of cAMP analogs. SR-BI mRNA and protein were expressed at high levels in the adrenal glands of knockout mice lacking the steroidogenic acute regulatory protein; these mice have extensive lipid deposits in the adrenocortical cells and high circulating levels of ACTH. Taken together, these studies suggest that trophic hormones can override the suppressive effect of cholesterol on SR-BI expression, thus ensuring that steroidogenesis is maintained during stress. PMID- 10478840 TI - Negative regulation of the androgen receptor gene promoter by NFI and an adjacently located multiprotein-binding site. AB - The upstream promoter of the rat androgen receptor (AR) gene contains a strong negative regulatory region located at the -388 to -340 nucleotide position. The distal part (-388/-373) of this regulatory region binds NFI, a ubiquitous transcription factor, while the proximal portion (-372/-340) contains an overlapping binding site for two nuclear proteins. This composite regulatory region (-388/-340) was initially defined by deoxyribonuclease I footprinting as the continuous stretch of a nuclease-protected site. NFI specificity of the distal portion (-388/-373) of the footprint was established through cross competition in electrophoretic mobility shift assay (EMSA) using the well characterized NFI element of the adenovirus major late promoter and by immunoreactivity to the NFI antibody. EMSA with oligonucleotide duplexes corresponding to the proximal domain (-372/-340) indicated multiple retarded bands with at least two major DNA-protein complexes. Further analysis with truncated oligonucleotide duplexes showed that these two major proteins bind to this domain in an overlapping manner. Within this overlapping area, the position spanning -359 to -347 is essential for the formation of either of these two complexes. Substitution of four G with T residues in the overlapping area totally abolished all protein binding at the downstream -372/-340 site. Point mutations that abolish specific binding at either the NFI or immediately downstream multiprotein-binding site caused about a 10-fold increase in AR promoter activity in transfected HepG2 cells. Double mutation involving both the NFI and proximal overlapping protein-binding sites failed to cause any additional increase in promoter function. From these results we conclude that the AR promoter contains a composite negative regulatory region at -388/-340, and the repressor function may involve a coordinate interaction between NFI and at least two other nuclear factors. PMID- 10478841 TI - The equine luteinizing hormone beta-subunit promoter contains two functional steroidogenic factor-1 response elements. AB - The requirements for basal expression of the LH beta-subunit promoter in pituitary gonadotropes are largely unknown. We have used the equine (e) LHbeta subunit promoter as a model to unravel the combinatorial code required for gonadotrope expression. Through the use of 5'-deletion mutagenesis, a region between -185 and -100 of the eLHbeta promoter was shown to play a critical role in maintaining basal promoter activity in alphaT3-1 and LbetaT2 cells. This region encompasses the steroidogenic factor-1 (SF-1) binding site that has been reported to have a functional role in expression of the LHbeta promoter in other species. We have also identified an additional SF-1 site at -55 to -48. Binding of SF-1 to both sites was confirmed by electrophoretic mobility shift assays. Mutations within these sites, either individually or in combination, did not attenuate basal activity of the eLHbeta promoter in alphaT3-1 cells, but did diminish promoter activity in LbetaT2 cells. Interestingly, cotransfection with an expression vector encoding SF-1 induced eLHbeta promoter activity, and this induction was abrogated by mutations within the SF-1 sites in alphaT3-1 cells. Block replacement mutagenesis was performed on the -185/-100 region of the eLHbeta promoter to identify DNA response elements responsible for maintaining basal promoter activity. From this analysis, two regions emerged as being important: a distal 31-bp segment (-181 to -150) and an element located immediately 3' to the distal SF-1 site (-119 to -106). It is hypothesized that these two regions as well as the SF-1 sites represent regulatory elements that contribute to a combinatorial code involved in targeting expression of the eLHbeta promoter to gonadotropes. PMID- 10478842 TI - Transcriptional activation of c-fos protooncogene by 17beta-estradiol: mechanism of aryl hydrocarbon receptor-mediated inhibition. AB - 17Beta-estradiol (E2) induced c-fos protooncogene mRNA levels in MCF-7 human breast cancer cells, and maximal induction was observed within 1 h after treatment. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) inhibited the E2-induced response within 2 h. The molecular mechanism of this response was further investigated using pFC2-CAT, a construct containing a -1400 to +41 sequence from the human c-fos protooncogene linked to a bacterial chloramphenicol acetyltransferase (CAT) reporter gene. In MCF-7 cells transiently transfected with pFC2-CAT, 10 nM E2 induced an 8.5-fold increase of CAT activity, and cotreatment with 10 nM TCDD decreased this response by more than 45%. Alpha Naphthoflavone, an aryl hydrocarbon receptor (AhR) antagonist, blocked the inhibitory effects of TCDD; moreover, the inhibitory response was not observed in variant Ah-nonresponsive MCF-7 cells, suggesting that the AhR complex was required for estrogen receptor cross-talk. The E2-responsive sequence (-1220 to 1155) in the c-fos gene promoter contains two putative core pentanucleotide dioxin-responsive elements (DREs) at -1206 to -1202 and -1163 to -1159. In transient transfection assays using wild-type and core DRE mutant constructs, the downstream core DRE (at -1163 to -1159) was identified as a functional inhibitory DRE. The results of photo-induced cross-linking, gel mobility shift, and in vitro DNA footprinting assays showed that the AhR complex interacted with the core DRE that also overlapped the E2-responsive GC-rich site (-1168 to -1161), suggesting that the mechanism for AhR-mediated inhibitory effects may be due to quenching or masking at the Sp1-binding site. PMID- 10478843 TI - Proteasome-mediated proteolysis of estrogen receptor: a novel component in autologous down-regulation. AB - Regulation of estrogen receptor (ER) concentration is a key component in limiting estrogen responsiveness in target cells. Yet the mechanisms governing ER concentration in the lactotrope cells of the anterior pituitary, a major site of estrogen action, are undetermined. In this study, we used a lactotrope cell line, PR1, to explore regulation of ER protein by estrogen. Estrogen treatment resulted in an approximate 60% decrease in ER steady state protein levels. Suprisingly, the decline in ER protein was apparent within 1 h of estrogen treatment and occurred in the absence of protein synthesis and transcription. Direct regulation of ER protein was further confirmed by pulse chase analysis, which showed that ER protein half-life was shortened from greater than 3 h to 1 h in the presence of estrogen. The estrogen-induced degradation of ER protein could be prevented by pretreatment with peptide aldehyde inhibitors of proteasome protease whereas inhibitors of calpain and lysosomal proteases were ineffective. Inhibition of proteasome activity maintained ER protein at a level equivalent to control cells not stimulated with estrogen but increased estrogen-binding activity by 1.75 fold. Proteolytic regulation of ER by the proteasome is not limited to pituitary lactotrope cells but is also operational in MCF-7 breast cancer cells, suggesting that this may be a common regulatory pathway used by estrogen. These studies describe a nongenomic action of estrogen that involves nuclear ER: rapid proteolysis of ER protein via a proteasome-mediated pathway. PMID- 10478844 TI - A truncated human peroxisome proliferator-activated receptor alpha splice variant with dominant negative activity. AB - The peroxisome proliferator-activated receptor alpha (PPARalpha) plays a key role in lipid and lipoprotein metabolism. However, important inter- and intraspecies differences exist in the response to PPARalpha activators. This incited us to screen for PPARalpha variants with different signaling functions. In the present study, using a RT-PCR approach a variant human PPARalpha mRNA species was identified, which lacks the entire exon 6 due to alternative splicing. This deletion leads to the introduction of a premature stop codon, resulting in the formation of a truncated PPARalpha protein (PPARalphatr) lacking part of the hinge region and the entire ligand-binding domain. RNase protection analysis demonstrated that PPARalphatr mRNA is expressed in several human tissues and cells, representing between 20-50% of total PPARalpha mRNA. By contrast, PPARalphatr mRNA could not be detected in rodent tissues. Western blot analysis using PPARalpha-specific antibodies demonstrated the presence of an immunoreactive protein migrating at the size of in vitro produced PPARalphatr protein both in human hepatoma HepG2 cells and in human hepatocytes. Both in the presence or absence of 9-cis-retinoic acid receptor, PPARalphatr did not bind to DNA in gel shift assays. Immunocytochemical analysis of transfected CV-1 cells indicated that, whereas transfected PPARalphawt was mainly nuclear localized, the majority of PPARalphatr resided in the cytoplasm, with presence in the nucleus depending on cell culture conditions. Whereas a chimeric PPARalphatr protein containing a nuclear localization signal cloned at its N-terminal localized into the nucleus and exhibited strong negative activity on PPARalphawt transactivation function, PPARalphatr interfered with PPARalphatr transactivation function only under culture conditions inducing its nuclear localization. Cotransfection of the coactivator CREB-binding protein relieved the transcriptional repression of PPARalphawt by PPARalphatr, suggesting that the dominant negative effect of PPARalphatr might occur through competition for essential coactivators. In addition, PPARalphatr interfered with transcriptional activity of other nuclear receptors such as PPARgamma, hepatic nuclear factor-4, and glucocorticoid receptor-alpha, which share CREB-binding protein/p300 as a coactivator. Thus, we have identified a human PPARalpha splice variant that may negatively interfere with PPARalphawt function. Factors regulating either the ratio of PPARalphawt vs. PPARalphatr mRNA or the nuclear entry of PPARalphatr protein should therefore lead to altered signaling via the PPARalpha and, possibly also, other nuclear receptor pathways. PMID- 10478845 TI - Coactivators for the orphan nuclear receptor RORalpha. AB - A mutation in the nuclear orphan receptor RORalpha results in a severe impairment of cerebellar development by unknown mechanisms. We have shown previously that RORalpha contains a strong constitutive activation domain in its C terminus. We therefore searched for mammalian RORalpha coactivators using the minimal activation domain as bait in a two-hybrid screen. Several known and putative coactivators were isolated, including glucocorticoid receptor-interacting protein 1 (GRIP-1) and peroxisome proliferator-activated receptor (PPAR)-binding protein (PBP/TRAP220/DRIP205). These interactions were confirmed in vitro and require the intact activation domain of RORalpha although different requirements for interaction with GRIP-1 and PBP were detected. Even in the absence of exogenous ligand, RORalpha interacts with a complex or complexes of endogenous proteins, similar to those that bind to ligand-occupied thyroid hormone and vitamin D receptors. Both PBP and GRIP-1 were shown to be present in these complexes. Thus we have identified several potential RORalpha coactivators that, in contrast to the interactions with hormone receptors, interact with RORalpha in yeast, in bacterial extracts, and in mammalian cells in vivo and in vitro in the absence of exogenous ligand. GRIP-1 functioned as a coactivator for the RORalpha both in yeast and in mammalian cells. Thus, GRIP-1 is the first proven coactivator for RORalpha. PMID- 10478846 TI - Androgen specificity of a response unit upstream of the human secretory component gene is mediated by differential receptor binding to an essential androgen response element. AB - The expression of secretory component (SC), the epithelial receptor for poly immunoglobulins, is regulated in a highly tissue-specific manner. In several tissues, e.g. lacrimal gland and prostate, SC synthesis is enhanced by androgens at the transcriptional level. In this study, we describe the presence of an androgen response unit, located 3.3 kb upstream of the sc transcription initiation site and containing several 5'-TGTTCT-3'-like motifs. Although each of these elements is implicated in the enhancer function, one element, the ARE1.2 motif, is found to be the main interaction site for the androgen receptor as demonstrated in in vitro binding assays as well as in transient transfection assays. A high-affinity binding site for nuclear factor I, adjacent to this ARE, is also involved in the correct functioning of the sc upstream enhancer. The ARE1.2 motif consists of an imperfect direct repeat of two core binding elements with a three-nucleotide spacer and therefore constitutes a nonconventional ARE. We demonstrate that this element displays selectivity for the androgen receptor as opposed to glucocorticoid receptor both in in vitro binding assays and in transfection experiments. Mutational analysis suggests that the direct nature of the half-site repeat is responsible for this selectivity. We have thus determined a complex and androgen-specific response unit in the far upstream region of the human SC gene, which we believe to be involved in its androgen responsiveness in epithelial cells of different organs such as prostate and lacrimal gland. We were also able to demonstrate that the primary sequence of a single nonconventional ARE motif within the enhancer is responsible for its androgen specificity. PMID- 10478847 TI - Two functionally different protein isoforms are produced from the chicken estrogen receptor-alpha gene. AB - The existence of two forms of the chicken estrogen receptor-alpha protein (ER alpha) in chicken tissues is demonstrated: the previously reported receptor (cER alpha form I), which has a size of 66 kDa, and a new form (cER-alpha form II), which lacks the N-terminal 41 amino acids present in form I and thus gives rise to a protein of 61 kDa. Whereas the 66-kDa protein is the translation product of several cER-alpha mRNAs (A1-D), the cER-alpha protein isoform II is encoded by a new cER-alpha mRNA (A2), which is transcribed in vivo from a specific promoter that is located in the region of the previously assigned translation start site of the cER-alpha gene. SI nuclease mapping analysis reveals that cER-alpha mRNA A2 is liver enriched. The resulting cER-alpha forms I and II differ in their ability to modulate estrogen target gene expression in a promoter- and cell type specific manner. Whereas cER-alpha form I activates or represses in a strictly E2 dependent manner, the truncated form is characterized by a partial transactivating or repressing activity in the absence of its ligand. Comparison of the N-terminal coding regions of different vertebrate ER-alpha reveal a conservation of the translation start methionine of the protein ER-alpha form II in other oviparous species but not in mammals. The expression of two classes of ER-alpha transcripts encoding the two ER-alpha receptor forms in the liver of Xenopus laevis and rainbow trout is demonstrated. Therefore, the existence of two functionally different protein isoforms produced from the ER-alpha gene is probably a common and specific feature in oviparous species. PMID- 10478848 TI - Function of steroidogenic factor 1 domains in nuclear localization, transactivation, and interaction with transcription factor TFIIB and c-Jun. AB - Normal endocrine development and function require nuclear hormone receptor SF-1 (steroidogenic factor 1). To understand the molecular mechanism of SF-1 action, we have investigated its domain function by mutagenesis and functional analyses. Our mutant studies show that the putative AF2 (activation function 2) helix located at the C-terminal end is indispensable for gene activation. SF-1 does not have an N-terminal AF1 domain. Instead, it contains a unique FP region, composed of the Ftz-F1 box and the proline cluster, after the zinc finger motif. The FP region interacts with transcription factor IIB (TFIIB) in vitro. This interaction requires residues 178-201 of TFIIB, a domain capable of binding several transcription factors. The FP region also mediates physical interaction with c Jun, and this interaction greatly enhances SF-1 activity. The putative SF-1 ligand, 25-hydroxycholesterol, has no effects on these bindings. In addition, the Ftz-F1 box contains a bipartite nuclear localization signal (NLS). Removing the basic residues at either end of the key nuclear localization sequence NLS2.2 abolishes the nuclear transport. Expression of mutants containing only the FP region or lacking the AF2 domain blocks wild-type SF-1 activity in cells. By contrast, the mutant having a truncated nuclear localization signal lacks this dominant negative effect. These results delineate the importance of the FP and AF2 regions in nuclear localization, protein-protein interaction, and transcriptional activation. PMID- 10478849 TI - Involvement of G protein-coupled receptor kinases and arrestins in desensitization to follicle-stimulating hormone action. AB - FSH rapidly desensitizes the FSH-receptor (FSH-R) upon binding. Very little information is available concerning the regulatory proteins involved in this process. In the present study, we investigated whether G protein-coupled receptor kinases (GRKs) and arrestins have a role in FSH-R desensitization, using a mouse Ltk 7/12 cell line stably overexpressing the rat FSH-R as a model. We found that these cells, which express GRK2, GRK3, GRK5, and GRK6 as well as beta-arrestins 1 and 2 as detected by RT-PCR and by Western blotting, were rapidly desensitized in the presence of FSH. Overexpression of GRKs and/or beta-arrestins in Ltk 7/12 cells allowed us to demonstrate 1) that GRK2, -3, -5, -6a, and -6b inhibit the FSH-R-mediated signaling (from 71% to 96% of maximal inhibition depending on the kinase, P < 0.001); 2) that beta-arrestins 1 or 2 also decrease the FSH action when overexpressed (80% of maximal inhibition, P < 0.01) whereas dominant negative beta-arrestin 2 [319-418] potentiates it 8-fold (P < 0.001); 3) that beta-arrestins and GRKs (except GRK6a) exert additive inhibition on FSH-induced response; and 4) that FSH-R desensitization depends upon the endogenous expression of GRKs, since there is potentiation of the FSH response (2- to 3 fold, P < 0.05) with antisenses cDNAs for GRK2, -5, and -6, but not GRK3. Our results show that the desensitization of the FSH-induced response involves the GRK/arrestin system. PMID- 10478850 TI - Signals from the AT2 (angiotensin type 2) receptor of angiotensin II inhibit p21ras and activate MAPK (mitogen-activated protein kinase) to induce morphological neuronal differentiation in NG108-15 cells. AB - In a previous study, we had shown that activation of the AT2 (angiotensin type 2) receptor of angiotensin II (Ang II) induced morphological differentiation of the neuronal cell line NG108-15. In the present study, we investigated the nature of the possible intracellular mediators involved in the AT2 effect. We found that stimulation of AT2 receptors in NG108-15 cells resulted in time-dependent modulation of tyrosine phosphorylation of a number of cytoplasmic proteins. Stimulation of NG108-15 cells with Ang II induced a decrease in GTP-bound p21ras but a sustained increase in the activity of p42mapk and p44mapk as well as neurite outgrowth. Similarly, neurite elongation, increased polymerized tubulin levels, and increased mitogen-activated protein kinase (MAPK) activity were also observed in a stably transfected NG108-15 cell line expressing the dominant negative mutant of p21ras, RasN17. These results support the observation that inhibition of p21ras did not impair the effect of Ang II on its ability to stimulate MAPK activity. While 10 microM of the MEK inhibitor, PD98059, only moderately affected elongation, 50 microM PD98059 completely blocked the Ang II- and the RasN17-mediated induction of neurite outgrowth. These results demonstrate that some of the events associated with the AT2 receptor-induced neuronal morphological differentiation of NG108-15 cells not only include inhibition of p21ras but an increase in MAPK activity as well, which is essential for neurite outgrowth. PMID- 10478852 TI - Some notes on the history of Haber's law. PMID- 10478851 TI - Toluene-3,4-dithiol analysis of blood for assessing carbon disulfide exposure. AB - Carbon disulfide is a neurotoxic compound used in the production of viscose rayon, and is a major decomposition product of dithiocarbamates used in industry, agriculture, and medicine. Methods used currently for assessing exposure to CS2 are limited in their ability to evaluate cumulative exposures and provide useful information for relatively short periods of time after exposure has ended. The present investigation evaluates a method for monitoring CS2 exposure that consists of cleaving the thiocarbonyl function of free CS2 or certain CS2 generated modifications on proteins using toluene-3,4-dithiol. The resulting toluene trithiocarbonate product is then quantified using reverse-phase high performance liquid chromatography. The sensitivity, dose response, kinetics and specificity of this biomarker in blood were examined in rats administered CS2 by inhalation, intraperitoneal injection, or gavage for acute through subchronic periods. Dithiol reactive functions in plasma and hemolysate demonstrated a linear dose response over a wide range of exposure levels, were dependent upon the duration of exposure, and appeared to have an appropriate sensitivity for evaluating occupational levels of exposure. Elimination rates of dithiol reactive functions may also be dependent upon exposure duration and exhibit different kinetics for plasma and hemolysate suggesting that elimination rates may be useful for estimating cumulative exposure and intervals between exposure and sample procurement. Dithiol analysis, used in conjunction with previously established erythrocyte protein cross-linking biomarkers, may provide a means to characterize the internal dose of CS2 resulting from acute through chronic periods, and may provide insight into the level of CS2-mediated covalent protein modifications occurring within the nervous system. PMID- 10478853 TI - The Single Exposure Carcinogen Database: assessing the circumstances under which a single exposure to a carcinogen can cause cancer. AB - A relational retrieval database has been developed compiling toxicological studies that assess whether a single dose of a chemical or physical agent, without exogenous promotional stimuli, could cause tumor development in animal models. This database allows for an evaluation of these studies over numerous parameters important to tumor outcome, which include type and quality of the studies as well as physical/chemical properties of the agents. An assessment of the database, which currently contains approximately 5500 studies involving about 800 chemicals from 2000 articles, reveals that a single dose of an agent can cause tumors to develop in males and females of numerous animal models in all principal age groups. In addition, the range of the 426 agents causing a positive response is chemically diverse, with representatives from over several dozen chemical classes. The dose caused a tumor endpoint was generally not acutely life threatening and was frequently a low proportion of the LD50 (i.e., less than 1/50 LD50). Positive responses also were reported via multiple routes of exposure, mainly oral, by injection, or dermal. These findings indicate that the phenomenon of single-exposure carcinogenesis is widespread and highly generalizable across chemical class, route, dose range, species, age, and gender. Single-exposure carcinogenesis, a concept long de-emphasized by regulatory agencies, requires a careful and formal consideration, especially as it may pertain to accidental spills, leaks, fires, explosions, and exposure excursions, but not necessarily limited to these. PMID- 10478854 TI - N-nitrosodiethylamine initiation of carcinogenesis in Japanese medaka (Oryzias latipes): hepatocellular proliferation, toxicity, and neoplastic lesions resulting from short term, low level exposure. AB - To investigate relationships among carcinogen exposure, cell proliferation, and carcinogenesis, 14-day post-hatch Japanese medaka (Oryzias latipes) were exposed to 0, 10, 25, 50, or 100 ppm N-nitrosodiethylamine (DEN) for 48 h under static renewal conditions. They were then held in clean water until sampling at 3 and 6 months. The frequencies of hepatic lesions and neoplasms were determined from hematoxylin/eosin-stained paraffin sections. A significant (p < 0.0001) concentration-related increase in hepatic vacuolated foci occurred in 3- and 6 month samples, with males having a significantly (p = 0.02) higher incidence than females. Concentration-related increases in degenerative lesions were documented for spongiosis hepatis at 3 months (p = 0.053) and hepatic vacuoles at 6 months (p = 0.005). There was a significant (p = 0.0001) concentration-related increase in macrophage aggregates at 6 months. Basophilic foci were significantly related (p < 0.0001) to DEN concentration at 3 months post-exposure and were unaffected by gender or age. At both 3 and 6 months, there were significant concentration related increases in hepatocellular carcinoma (p < or = 0.02). Hepatocyte proliferation in 3-month whole specimens was quantified using an immunohistochemical assay for proliferating-cell nuclear antigen. Trend tests and a probit dose-response model showed a significantly positive correlation (p = 0.015) between proliferating hepatocytes and DEN concentrations. These results confirm that short-term exposure to low and moderate levels of DEN initiates concentration-dependent carcinogenic effects in medaka that are apparent at 3 months postexposure. DEN could be an effective initiator in an initiation/promotion assay for medaka using a 48-h exposure period, DEN concentrations < or = 10 ppm, and a 6-month sampling period. PMID- 10478855 TI - Chronic peroxisome proliferation and hepatomegaly associated with the hepatocellular tumorigenesis of di(2-ethylhexyl)phthalate and the effects of recovery. AB - This study compared the levels of cell proliferation and peroxisome proliferation in rodent liver with tumor incidence, to provide more information on the relationship between these events following chronic exposure. Fischer 344 rats were treated with 0, 100, 500, 2500, or 12,500 ppm DEHP, and B6C3F1 mice were treated with 0, 100, 500, 1500, or 6000 ppm DEHP in the diet for up to 104 weeks. Additional groups of rats and mice received the highest concentration for 78 weeks and then the control diet for an additional 26 weeks (recovery groups). Animals were terminated at weeks 79 and 105 for histopathologic examination. Elevated palmitoyl CoA oxidation activity and higher liver-to-body weight ratios were observed for the 2500- and 12,500-ppm groups of rats, and for the 500-, 1500 , and 6000-ppm groups of mice at Week 105. No increase in palmitoyl CoA oxidation activity was evident in the recovery group, and relative liver weights were near control levels following recovery. No hepatic cell proliferation was detected at Weeks 79 or 105 in either species although preliminary data indicated that cell proliferation did occur within the first 13 weeks of exposure. A significantly higher incidence of hepatocellular tumors was only observed for the 2500- and 12,500-ppm group and its recovery group of rats, and for the 500-, 1500-, and 6000-ppm groups and the recovery group of mice. The tumor incidences were reduced for the recovery groups compared with the groups fed DEHP continuously for 104 weeks. The data indicate that high levels of peroxisome proliferation and hepatomegaly are associated with DEHP hepatocarcinogenesis in rodent liver, and that the tumorigenic process may be arrested by cessation of DEHP treatment, suggesting that extended treatment with DEHP acts to promote tumor growth. PMID- 10478856 TI - Estrogenic potencies of several environmental pollutants, as determined by vitellogenin induction in a carp hepatocyte assay. AB - Estrogenic potencies of several xenoestrogens were determined in vitro, using cultured hepatocytes from a genetically uniform male carp strain (Cyprinus carpio). Estrogenicity was measured as induction of the yolk protein precursor vitellogenin (Vtg), and compared to Vtg induction by 17beta-estradiol (E2). The order of estrogenic potency was: methoxychlor (MXCL) > o,p-DDT > chlordecone approximately/= bisphenol-A approximately/= 4-t-pentylphenol. Estrogenic potencies of these compounds varied from 1 x 10(-3) to 1 x 10(-4) relative to E2. The synthetic estrogen DES had a relative estrogenic potency of 0.5, whereas dieldrin, beta-endosulfan, o,p-DDE, and toxaphene (technical mixture) did not induce vitellogenesis at concentrations up to 100 microM. Experiments in which cells were simultaneously exposed to E2 and these xenoestrogens showed that the Vtg-inducing activities of E2 and 4-t-pentylphenol or bisphenol-A were (partially) additive, whereas E2 antagonized the estrogenic effects of MXCL and o,p-DDT. The effect of cytochrome P4501A (CYP1A)-induction on the estrogenicity of MXCL was studied by co-exposing cells to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). TCDD (10 pM) caused a greater than 50-fold induction of CYP1A, measured as ethoxyresorufin O-deethylase (EROD) activity, but Vtg induction by MXCL was not significantly affected. This indicates that CYP1A is not involved in the bioactivation of MXCL to more potent estrogenic metabolites in carp. The CARP-HEP (hepatocyte) assay can detect xenoestrogens with a potency > or = 2 x 10(-5) relative to E2. It allows simultaneous testing of more than 10 compounds for both estrogenic and antiestrogenic effects, which makes it a promising tool for the screening of suspected xenoestrogens. PMID- 10478857 TI - Detection of immunotoxicity of benzo[a]pyrene in a subacute toxicity study after oral exposure in rats. AB - In an extended OECD 407 study protocol, including immune parameters, male Riv:Tox Wistar SPF rats were treated for 35 days with benzo[a]pyrene (B[a]p) (3, 10, 30, or 90 mg/kg body weight) by gavage. Oral administration of B[a]p in rats resulted not only in general toxicity, as indicated by the effects on body weight, but also in immunotoxicity, as indicated by the effects on bone marrow, thymus, spleen, and lymph nodes. Oral B[a]p induced a dose-related decrease in thymus weight (at 10, 30, and 90-mg/kg). Lymph node weights (popliteal, mandibular, and mesenteric) were decreased in the 90-mg/kg rats only. Histologically, indications for cortical atrophy were noted in the thymuses of the 30- and 90-mg/kg dose groups, which was confirmed by morphometric analysis. Nucleated spleen and bone marrow cell counts were decreased in the 90-mg/kg group. Both the absolute number (90 mg/kg) and relative number (10, 30, and 90 mg/kg) of B cells in the spleen were decreased. Red blood cell (RBC) and white blood cell (WBC) counts were significantly decreased; for the WBC at 90 mg/kg, and for the RBC at 10, 30, and 90 mg/kg. The absolute number of lymphocytes and eosinophilic granulocytes was decreased in the 90-mg/kg group, while the absolute number of monocytes was increased in the 10- and 30-mg/kg dose groups. Serum immunoglobulin levels showed a decrease of IgM and IgA after treatment of the animals with 30 and 90 mg/kg, respectively. The highest dose of B[a]p treatment (90 mg/kg) resulted in a significant decrease of natural killer (NK)-cell activity in the spleen. Most toxic effects were only observed in the highest-dose group (90 mg/kg), but compared to the general toxicity, some parameters indicating immunotoxic effects were also affected at lower doses (10 and 30 mg/kg). In conclusion, immunotoxicity of B[a]p can be detected using parameters of the immune system such as described in the recently updated OECD 407 guideline. In the present study thymus weight changed and spleen B-cell populations were affected at a dose of 10 mg/kg, a level where no overt general toxicity was noted. PMID- 10478858 TI - Metallothionein protects against the nephrotoxicity produced by chronic CdMT exposure. AB - Metallothionein (MT) is a low-molecular-weight, cysteine-rich, metal-binding protein. Induction of MT has been proposed to be an important adaptive mechanism in decreasing Cd toxicity. MT has been shown to protect against CdCl2-induced lethality and hepatotoxicity; however, MT does not protect against acute CdMT induced nephrotoxicity. This study was aimed at clarifying the role of metallothionein in chronic CdMT-induced renal injury. Wild type and MT-I/II knockout (MT-null) mice were therefore given sc injections of CdMT (25 and 100 microg Cd/kg) or saline daily, 6 times/week for 6 weeks, and renal injury was evaluated. Multiple injections of CdMT to wild-type mice resulted in renal Cd concentrations up to 120 microg/g kidney, along with a 100-fold increase in renal MT (450 microg/g kidney). In contrast, renal Cd concentration in MT-null mice administered multiple injections of CdMT reached a much lower level than in wild type mice (<10 microg/g kidney). Although less Cd accumulated in their kidneys, MT-null mice were more susceptible than wild-type mice to CdMT-induced nephrotoxicity, as indicated by increased urinary excretion of protein and N acetyl-beta-D-glucosaminidase, as well as by elevated blood urea nitrogen levels. At the higher daily dose of CdMT (100 microg Cd/kg), kidneys of MT-null mice were enlarged. Chronic CdMT administration eventually damaged the entire kidney, which included glomerular swelling, interstitial inflammation, edema, tubular cell degeneration, and atrophy. In contrast to a single injection of CdMT that produces proximal tubular necrosis, chronic injection of CdMT results in tubular cell apoptosis in both wild-type and MT-null mice. These data indicate that chronic CdMT administration produces similar renal injury to that observed after chronic CdCl2 administration, and that intracellular MT protects against nephrotoxicity produced by chronic CdMT administration. PMID- 10478859 TI - Bioactivation of cyanide to cyanate in sulfur amino acid deficiency: relevance to neurological disease in humans subsisting on cassava. AB - Neurological disorders have been reported from parts of Africa with protein deficient populations and attributed to cyanide (CN-) exposure from prolonged dietary use of cassava, a cyanophoric plant. Cyanide is normally metabolized to thiocyanate (SCN-) by the sulfur-dependent enzyme rhodanese. However, in protein deficient subjects where sulfur amino acids (SAA) are low, CN may conceivably be converted to cyanate (OCN-), which is known to cause neurodegenerative disease in humans and animals. This study investigates the fate of potassium cyanide administered orally to rats maintained for up to 4 weeks on either a balanced diet (BD) or a diet lacking the SAAs, L-cystine and L-methionine. In both groups, there was a time-dependent increase in plasma cyanate, with exponential OCN- increases in SAA-deficient rats. A strongly positive linear relationship between blood CN- and plasma OCN- concentrations was observed in these animals. These data are consistent with the hypothesis that cyanate is an important mediator of chronic cyanide neurotoxicity during protein-calorie deficiency. The potential role of thiocyanate in cassava-associated konzo is discussed in relationship to the etiology of the comparable pattern of motor-system disease (spastic paraparesis) seen in lathyrism. PMID- 10478861 TI - Lead acetate exposure inhibits nitric oxide synthase activity in capillary and synaptosomal fractions of mouse brain. AB - The toxicity of lead (Pb) is of concern to public health due to its persistence in the environment. Brain is one of the major target organs where severe neurologic alterations may be triggered after exposure. The primary effects of lead on brain functions are thought to be a damage to the nervous system microvasculature. However, the mechanism of this toxicity is poorly understood. Nitric oxide synthase (NOS) may be a target for lead and changes in its function can result in a cascade of pathophysiological effects that may be observed in isolated capillaries and synaptosomes. We have determined the concentration of lead in blood, capillaries and synaptosomes in brain from mice receiving 0, 250, 500, and 1000 ppm of lead for 14 days, through the drinking water. NOS activity was determined in the capillaries and synaptosomes by following the conversion of 3H-L-arginine to 3H-L-citrulline. The results show that blood lead levels were dose-dependent. Brain capillaries showed a preferential accumulation of lead as compared to synaptosomes. With all Pb treatments, synaptosomal constitutive NOS was inhibited (about 50% of control) while the inducible NOS activity in capillaries was enhanced. These data suggest that inhibition of cNOS activity and increase in iNOS may contribute to the Pb effects on the CNS. PMID- 10478860 TI - Differential ability of astroglia and neuronal cells to accumulate lead: dependence on cell type and on degree of differentiation. AB - The apparent ability of astroglia to serve as a lead (Pb) sink in the mature brain may result from either their strategic location, between the blood-brain barrier and neurons, or from intrinsic differences between the ability of astroglia and neurons to accumulate this metal. This phenomenon may be dependent on the degree of cell differentiation. In order to address the latter possibility, Pb accumulation was compared among the following cell culture models: (1) mature and immature rat astroglia, (2) undifferentiated SY5Y human neuroblastoma cells and SY5Y cells differentiated with nerve growth factor, (3) immature rat astroglia grown in differently conditioned media, some of which induce partial differentiation, and (4) rat astroglia and SY5Y cells in co culture. Astroglial cultures, prepared from 1-day-old rat cerebral hemispheres, were exposed to 1 microM Pb after either 14 (immature) or 21 (mature) days in culture. Pb content of the cells was measured by atomic absorption spectroscopy. Immature astroglia took up less Pb when glutathione (GSH) was added to the medium, suggesting that GSH may regulate Pb uptake in these cells. Undifferentiated neuroblastoma cells accumulated more Pb than did the differentiated ones. Astroglia accumulated up to 24 times more Pb than did neuronal cells. This ability was enhanced by exposure to conditioned medium from a neuroblastoma cell line, but not by endothelial cell-conditioned medium, although this medium induced the expression of a glutamate-activated Ca2+ response. Our findings are in agreement with in vivo studies, and thus validate the use of these cell-culture models for future studies on differential mechanisms of Pb uptake. PMID- 10478862 TI - Assessment of adult and neonatal reproductive parameters in Sprague-Dawley rats exposed to propylene glycol monomethyl ether vapors for two generations. AB - This study evaluated propylene glycol monomethyl ether (PGME) in a rat 2 generation reproduction study, which included non-traditional study end points, such as sperm count and motility, developmental landmarks, estrous cyclicity, and weanling organ weights. Groups of 30 male and 30 female Sprague-Dawley rats (6 weeks-old) were exposed to 0, 300, 1000, or 3000 ppm of PGME vapors via inhalation for 6 hours/day, 5 days/week prior to mating, and 6 hours/day, 7 days/week during mating, gestation, and lactation, for 2 generations. These concentrations corresponded to estimated oral equivalent doses of 0, 396, 1325, or 3974 mg/kg/day. At 3000 ppm, toxicity in the P1 and P2 adults was marked, as evidenced by sedation during and after exposure, and mean body weights which were as much as 21% lower than controls. This marked parental toxicity was accompanied by lengthened estrous cycles, decreased fertility, decreased ovary weights, and histologic ovarian atrophy in maternal rats. In the offspring from these dams, decreased body weights, reduced survival and litter size, slight delays in puberty onset, and histologic changes in liver and thymus in the F1 and F2 offspring were observed. The nature of the reproductive/neonatal effects and their close individual animal correlation with decreased maternal body weights suggested that these effects were secondary to general toxicity and/or nutritional stress. No such reproductive/neonatal effects were observed at 1000 ppm, a concentration which caused less marked, but significant body weight effects without sedation. There were no treatment-related effects of any kind noted at 300 ppm of PGME. Therefore, the no-observable-effect level (NOEL) for reproductive/neonatal effects was 1000 ppm, and that for parental toxicity was 300 ppm. PMID- 10478863 TI - Dose-rate effects of ethylene oxide exposure on developmental toxicity. AB - In risk assessment, evaluating a health effect at a duration of exposure that is untested involves assuming that equivalent multiples of concentration (C) and duration (T) of exposure have the same effect. The limitations of this approach (attributed to F. Haber, Zur Geschichte des Gaskrieges [On the history of gas warfare], in Funf Vortrage aus den Jahren 1920-1923 [Five lectures from the years 1920-1923], 1924, Springer, Berlin, pp. 76-92), have been noted in several studies. The study presented in this paper was designed to specifically look at dose-rate (C x T) effects, and it forms an ideal case study to implement statistical models and to examine the statistical issues in risk assessment. Pregnant female C57BL/6J mice were exposed, on gestational day 7, to ethylene oxide (EtO) via inhalation for 1.5, 3, or 6 h at exposures that result in C x T multiples of 2100 or 2700 ppm-h. EtO was selected because of its short half-life, documented developmental toxicity, and relevance to exposures that occur in occupational settings. Concurrent experiments were run with animals exposed to air for similar periods. Statistical analysis using models developed to assess dose-rate effects revealed significant effects with respect to fetal death and resorptions, malformations, crown-to-rump length, and fetal weight. Animals exposed to short, high exposures of EtO on day 7 of gestation were found to have more adverse effects than animals exposed to the same C x T multiple but at longer, lower exposures. The implication for risk assessment is that applying Haber's Law could potentially lead to an underestimation of risk at a shorter duration of exposure and an overestimation of risk at a longer duration of exposure. Further research, toxicological and statistical, are required to understand the mechanism of the dose-rate effects, and how to incorporate the mechanistic information into the risk assessment decision process. PMID- 10478864 TI - Maternal and developmental toxicity evaluation of melatonin administered orally to pregnant Sprague-Dawley rats. AB - Melatonin (MEL) is a widely used, over-the-counter sleep aid, and it has putative contraceptive, antioxidant, antiaging, and anticancer effects. The developmental toxicity potential for repeated oral doses of MEL had not previously been evaluated. In the present studies, time-mated, Sprague-Dawley-derived (CD) rats were administered MEL or vehicle by gavage on gestation days (gd) 6-19. MEL treated groups received 1-, 10-, 100-, 150-, or 200-mg/kg body weight/day in the screening study (15 rats/group), and 50, 100, or 200 mg/kg/day in the definitive study (25 rats/group). In both studies, maternal food/water consumption, body weight, and clinical signs were monitored at regular intervals throughout gestation. At termination (gd 20, both studies), maternal liver and gravid uterine weights, number of ovarian corpora lutea, conceptus survival, fetal sex, and fetal body weight were evaluated. Fetal morphological examination included external structures (both studies) as well as visceral and skeletal structures (definitive study). In the screening study, maternal serum levels of 17beta estradiol, progesterone, prolactin, and luteinizing hormone were determined by radioimmunoassay, and mammary tissue was fixed, stained, and evaluated for percent glandular area within the fat pad. No maternal morbidity/mortality was found in either study. In the screening study, aversion to treatment (> or =100 mg/kg/day) and reduced maternal weight gain (> or =150 mg/kg/day) were noted, but reproductive/endocrine parameters and fetal development were not affected. In the definitive study, aversion to treatment was noted at > or =50 mg/kg/day, and mild sedation, reduced maternal food intake, and reduced body weight gain were found during initial treatment with 200 mg/kg/day. MEL had no effect on prenatal survival, fetal body weight, or incidences of fetal malformations/variations. Thus, in the definitive study, the maternal toxicity NOAEL and LOAEL were 100 and 200 mg/kg/day, respectively, and the developmental toxicity NOAEL was > or =200 mg/kg/day. PMID- 10478866 TI - Differential sensitivity of lung and brain to sulfide exposure: a peripheral mechanism for apnea. AB - Acute exposure to hydrogen sulfide (H2S) causes 4 dose-dependent responses: hyperpnea, unconsciousness or knockdown, apnea, and death. At present, scientific mechanisms for these effects are unknown, but inhibition of cytochrome oxidase in the central nervous system (CNS) by sulfide has been suggested. In this study, the premise of brain as target is examined by comparing peripheral with direct delivery of sulfide to brain. NaHS was administered to anesthetized Sprague Dawley male rats, by femoral intravenous, or carotid intra-arterial injection. With apnea as the test, ED100 values of 0.6 and 3.0 mg kg(-1) were found for peripheral and direct delivery to brain, respectively. Peripheral delivery is 5 times as effective as direct delivery to the brain, and greater if the relative dilution factors are considered. Lidocaine, applied to the vagus, prevented apnea, so lung-brain transmission was essential. It was therefore concluded that the lung is the peripheral site of action. Hyperpnea increased in duration with dose. The carotid body, located distal to the arterial catheter, sensed the high doses of NaHS but did not trigger apnea. Intravenous infusion or intraperitoneal injection with NaHCO3 prevented hyperpnea, apnea, and death. The following conclusions are drawn: the carotid body is not implicated in NaHS-induced apnea, the lung and not brain harbors the primary site of action of H2S, and an afferent neural signal from the lung via the vagus induces the apnea. Finally, NaHCO3 appears to prevent toxicity from sulfide, and therefore H2S, by some unexplained mechanism. Practical emergency treatment for acute toxicity in the field may be possible. PMID- 10478865 TI - Susceptibility to chlordecone-carbon tetrachloride induced hepatotoxicity and lethality is both age and sex dependent. AB - The present paper examines the susceptibility to chlordecone (Kepone, CD) and carbon tetrachloride across different ages (35, 45, and 63-days-old) in male and female Sprague-Dawley rats using different lengths of time on a CD diet (10 ppm). The principal findings are that the hepatotoxicity and mortality associated with CD-CCl4 interaction is highly age-dependent for both sexes. There was marked hepatotoxicity occurring in both sexes as they reached 45 days-of-age and females were considerably more susceptible than males to both CD-CCl4-induced hepatotoxicity and lethality. While 63-day-old females are more susceptible to the CD-CCl4 interaction than their male counterparts, the magnitude of the sex difference is diminished from that observed in 45-day-old rats. These findings challenge the hypothesis of Mehendale (1990, Med. Hypotheses 33, 289-299) that chlordecone (CD) pretreatment eliminates the well-established sex difference in CCl4-treated rats. In contrast to the CD-CCl4 findings, the sex difference in CCl4-induced hepatotoxicity was not age-dependent and was consistent over the three ages studied. The findings that CD-CCl4 interaction is highly age-dependent (within the 3 ages tested) but that CCl4-induced hepatotoxicity is not, suggest that the CD-CCl4 interaction acts via a mechanism that does not primarily involve CCl4 potentiation. PMID- 10478867 TI - A gastric sexology: the story of three men and three bottles of gastric juice. PMID- 10478868 TI - More evidence of the failure of oral cyclosporine to maintain remission due to intravenous cyclosporine in IBD. PMID- 10478869 TI - Treating biopsies to cure patients: the management of histological findings in mucosa-associated lymphoid tissue (MALT) PMID- 10478870 TI - The GASTER project: building a computer network in digestive endoscopy: the experience of the European Society for Gastrointestinal Endoscopy. Gastrointestinal Endoscopy Application for Standards in Telecommunication, Education and Research. AB - Digestive endoscopy is currently the main diagnostic procedure for investigation of the digestive tract whenever a digestive disease is suspected. From 1970 to 1985, digestive endoscopy was performed with endoscopes equipped with fiberoptic bundles, whereas the last decade was marked by the development of electronic endoscopes, characterized by the presence of a CCD (charge coupled device) at the tip of the endoscope. Thus the physician looks at a TV screen to control the procedure and examine in detail the gut wall. Endoscopes examine the foregut until the duodenum and the hindgut, up to the three last intestinal loops. When the endoscopic workstation comprises a computer, it is possible to acquire electronic images during the endoscopy and use these images as support of the information about the results of the procedure. These numeric images can then be stored in databases containing text attached to them. Starting with these images, one may expect many developments in the near future that will change the management of the patient with digestive diseases. Physicians will become able to exchange images and text related to one patient or one procedure, although they are equipped with different workstations. Therefore, it is obvious that the information exchanged must be written in a standard format that makes it understandable by all systems. The European Society of Gastrointestinal Endoscopy is a scientific society that groups most of the gastroenterologists in Europe. This society has initiated a research program to develop standards for the exchange of images and text. The Gastrointestinal Endoscopy Applications for Standards in Telecommunication, Education, and Research (GASTER) project intends to implement a multimedia database of endoscopic images based on a standard format of images and a standard terminology for descriptive terms. These standards must be validated by use in different endoscopy units. The database will collect images from these centers that will be linked to the coordinating center through a network based on an integrated services digital network (fast electronic connection). This database will then be used for the development of computer applications. The output of the GASTER project will bring advances at three levels: (1) The physicians will be able to exchange images about the procedures their patients have undergone and will thus obtain more complete information, improving quality of care. They will also benefit from help-to decision applications based on validated reference images from the database. (2) At the patient level, the quality of care will be improved through a better dissemination of information between the physicians in charge of the patient, thus there is better follow-up of the patient and a decrease in redundant examinations. (3) At the level of national health care systems, the benefit will be a decrease in cost of care due to a better follow-up of the patients, a decrease in redundant examinations, and a faster decision made to treat the patient. The possibility of consulting a database of a scientifically validated images used as reference material will also improve quality control in digestive endoscopy. PMID- 10478871 TI - Partial fundoplications for gastroesophageal reflux disease: indications and current status. AB - The Nissen fundoplication, and in particular the laparoscopic Nissen fundoplication, has received widespread acceptance as the most definitive therapy for gastroesophageal reflux disease. There remains, however, certain patients who do better with a less aggressive surgical augmentation of the lower esophageal sphincter. Partial fundoplications originated in the early 1960s as an alternative procedure to the Nissen, which was associated with moderately high rates of postoperative side effects. These "more physiologic" procedures have proved successful in the treatment of reflux disease in patients with poor or no esophageal motility. In particular, the use of partial fundoplications in association with Heller's myotomy for achalasia has been demonstrated to be well tolerated and to reduce the risk of late dysphasia resulting from uncontrolled gastroesophageal reflux (GER). The use of partial fundoplications in GER patients with normal motility, however, has been less successful. High recurrence rates are documented by many centers with the main cause appearing to be related to a less competent neo-lower esophageal sphincter and a higher rate of wrap herniation. This has led to the current practice of a "tailored approach" to reflux disease, in which all patients receive a thorough preoperative physiologic evaluation to determine the best antireflux procedure for the individual. This is generally a Nissen repair for those with normal motility and either an extrashort "floppy" Nissen or a partial wrap for those with impaired peristalsis. PMID- 10478872 TI - Acquired gastric mucosa-associated lymphoid tissue (MALT): a review with special emphasis on association with extragastric diseases and management problems of gastric MALT. AB - Mucosa-associated lymphoid tissue is absent physiologically in normal gastric mucosa, but it can develop in some pathologies (dyspepsia, celiac disease, autoimmune diseases) due to certain (Helicobacter pylori, gluten) or uncertain (viruses?) antigenic stimuli. Its importance is related to the consideration that it is the background for the development of gastric mucosa-associated lymphoid tissue (MALT) lymphoma, and its histologic progression to MALT lymphoma may be diagnosed using Isaacson's score. Monoclonal bands can be diagnosed in acquired gastric MALT as well as in gastric MALT lymphoma, but its role is unknown. To prevent development of MALT lymphoma from acquired MALT, currently anti-H. pylori treatment in all H. pylori- and MALT-positive patients is not suggested--only in patients with grade 3 of acquired MALT--due to the high risk of developing MALT lymphoma in these patients. At the same time, the etiology of gastric MALT in autoimmune extragastric diseases is not clear. Additional studies are needed to clarify this relation and its possible causes. PMID- 10478873 TI - Cystic fibrosis and the pancreas: recent scientific advances. AB - Cystic fibrosis (CF) is inherited as an autosomal recessive trait due to a mutated gene carried on the long arm of chromosome 7. The protein coded by the CF gene (CFTR) is an apical chloride channel that regulates active chloride transport across epithelial cell membranes. The role of CFTR in pancreatic exocrine secretion is based on the concept that CFTR is localized to the apical membrane of the proximal duct epithelial cells. The primary ductal cell chloride channel abnormality results in dehydrated protein-rich secretions that obstruct the proximal ducts, leading to secondary acinar cell destruction, fibrosis, and exocrine pancreatic insufficiency in 85% of the CF population. A strong correlation between genotype and pancreatic phenotype is recognized. Advances in molecular biology such as cloning of lipase genes and gene therapy are challenging the field of pancreatic enzyme supplementation therapy. PMID- 10478874 TI - D-xylose testing. AB - The literature on D-xylose testing has been reviewed, stressing advances in our understanding of absorption in general (including D-xylose absorption), the relationship of D-xylose testing to the development of excellent serologic tests for the diagnosis of celiac disease, the use of D-xylose testing in the evaluation of diarrhea in acquired immunodeficiency syndrome, new information on breath testing for the evaluation of malabsorption, and recent information on the understanding of D-xylose absorption compared with transcellular vs. paracellular transport. The authors suggest ways in which D-xylose testing might be employed in malabsorption or diarrhea evaluations, including some algorithms. PMID- 10478875 TI - Cyclosporine therapy in inflammatory bowel disease: short-term and long-term results. AB - Intravenous cyclosporine therapy followed by oral cyclosporine therapy reduce the need for urgent surgery in steroid-refractory inflammatory bowel disease (IBD). Our objective is to report short- and long-term results of cyclosporine therapy in IBD patients. Thirteen patients with steroid-refractory IBD, seven patients with ulcerative colitis (UC), and six patients with Crohn's disease (CD) were treated with intravenous cyclosporine (4 mg/kg/day) for a mean period of 11.4+/ 2.8 days (range, 4-15 days). Subsequently the patients were started on oral cyclosporine (8 mg/kg/day) and followed for a mean of 10.3+/-10 months (range, 1 30 months). Twelve patients responded to intravenous cyclosporine therapy. One patient with UC developed sepsis on the fourth day of intravenous cyclosporine therapy and needed urgent colectomy. Nine of 12 initial responders (6 patients with UC and 3 patients with CD) relapsed during follow-up despite oral cyclosporine and underwent elective surgery. One patient with CD relapsed 3 months after discontinuation of oral cyclosporine. Only two patients with CD are in long-term remission. There were no long-term side effects in any of the 13 treated patients. In conclusion, intravenous cyclosporine was effective in inducing remission or significant improvement in 12 of 13 patients with steroid refractory IBD. However, with subsequent oral cyclosporine the remission could be maintained only for a short while. Each of the six patients with UC needed colectomy and three of the five patients with CD had intestinal resection within 12 months despite oral cyclosporine therapy. PMID- 10478876 TI - The role of transjugular liver biopsy in a liver transplant center. AB - Transjugular liver biopsy is a particularly useful procedure in overcoming the classic limitations of percutaneous biopsy, such as hemorrhagic diathesis or tense ascites. The authors evaluated the impact that this technique had in their liver transplant program, considering performance and safety in 160 procedures. Histologic characterization was accomplished in 75% of cirrhotic patients and in 96% of patients with acute liver disease. Confirmation of a presumptive diagnosis was made in 76% of patients and a previously unsuspected diagnosis was raised in 11%. Hemochromatosis, Wilson's disease, and acute alcoholic hepatitis were the most challenging diagnoses and, together with patients with liver failure, had the most relevant implications in transplant decisions. The availability of transjugular liver biopsy provided decisive information regarding patient selection and the best timing to proceed with transplantation. PMID- 10478877 TI - Esophageal variceal ligation by reloading with inexpensive hemorrhoidal O-ring- is an overtube necessary? AB - The overtube is the major cause for severe complications during endoscopic variceal ligation with a single-shot ligator. This retrospective study was designed to examine the necessity of the placement of an overtube during elective endoscopic variceal ligation. Thirty-one sessions in 18 patients were analyzed. An overtube was inserted using an over-the-scope technique in 11 sessions (group 1) but was omitted in 20 sessions (group II). The complications, technical difficulties, and operating time were analyzed. Child's grading, the size of the esophageal varices, and the number of rubber bands deployed were comparable in both groups. There was a significantly longer operating time (p < 0.01) and more oropharyngeal injury (p = 0.03) in group I than in group II. Mid esophageal injury, which was associated with resistance in withdrawing the gastroscope from the overtube, occurred in 55% of sessions in group I but in 0% of session in group II. In conclusion, the use of an overtube is associated with more complications, and it can be omitted during elective endoscopic variceal ligation. PMID- 10478878 TI - Tuberculosis of the bile duct: a rare cause of obstructive jaundice. AB - Tuberculosis of the bile duct is extremely rare. Patients with this condition usually present with a protracted illness and obstructive jaundice, which may be confused with hepatobiliary malignancies. A retrospective review of hospital records of patients who presented with tuberculosis of the bile duct between January 1986 and December 1996 was undertaken, and data were obtained concerning clinical presentation, investigations, treatment, and follow-up. Four patients (one man and three women) with a mean age of 44.8 years had tuberculosis of the bile duct. Diagnostic imaging techniques showed bile duct dilation in all four patients. Endoscopic retrograde cholangiopancreatography (ERCP) showed a stricture in the proximal common bile duct (CBD) in one patient, a stricture in the distal CBD in one patient, a stricture in the common hepatic duct (CHD) in one patient, and multiple strictures in the CHD and left intrahepatic duct in one patient. Bile cytology and fine-needle aspiration identified correctly the diagnosis in each patient. Two patients underwent laparotomy with the initial suspicion of cholangiocarcinoma; the correct diagnosis was made based on frozen sections taken intraoperatively. One patient was treated with endoscopic stenting and three patients underwent laparotomy for bile duct obstruction. All patients received antituberculous therapy. There were no deaths; all patients remained healthy at a mean follow-up of 36.5 months. It is important to obtain a tissue diagnosis in all patients with obstructive jaundice to avoid missing this rare but curable disease. The treatment of tuberculosis of the bile duct involves relief of the bile duct obstruction and antituberculous therapy. PMID- 10478879 TI - Demographic predictors of resource utilization for bleeding peptic ulcer disease: the Ontario GI Bleed Study. AB - Peptic ulcer disease (PUD) and its complications impose a substantial burden on health care resources. To help identify subpopulations in which preventative measures might achieve maximal cost savings, the authors studied the variation in resource utilization and cost for management of bleeding PUD among demographic subgroups. Resource utilization profiles and direct medical cost estimates were generated for consecutive admissions for bleeding PUD at four hospitals in southern Ontario via chart review and adaptation of an administrative cost database. Multiple linear regression models were developed to identify independent demographic predictors of direct medical case cost and hospital length of stay (LOS). Among 158 admissions, the average LOS and case costs were 5.73 days and $2,953 (Canadian) respectively. Age, comorbid illness, nonsteroidal anti-inflammatory drug use, and the absence of prior PUD or upper gastrointestinal hemorrhage were associated with higher cost in univariate analysis, whereas increasing age and comorbidity predicted LOS. Only age and absence of prior PUD persisted as independent predictors of direct medical cost and LOS in a stepwise multiple linear regression. Costs for managing bleeding PUD vary substantially among demographic subgroups. More careful stratification of treatment costs is needed when economic models of interventions to prevent or to treat PUD are applied to specific subpopulations. PMID- 10478880 TI - Gastrointestinal motor dysfunction, symptoms, and neuropathy in noninsulin dependent (type 2) diabetes mellitus. AB - Although relatively frequent. diabetic involvement of digestive tract motility has not been investigated extensively in different organs. The authors studied esophageal, gastric, and gallbladder motor function in 35 type 2 (noninsulin dependent) diabetic patients to determine the extent of gut involvement. Of these patients, 27 (77%) had peripheral neuropathy, 12 (34%) had both peripheral and autonomic neuropathy, and 22 (63%) had gastrointestinal symptoms. Esophageal manometric abnormalities were recorded in 18 patients, and delayed radionuclide emptying of the esophagus was documented in 16 patients, with a 83% concordance between the two tests. Scintigraphic gastric emptying of solids was delayed in 56% of patients, whereas gallbladder emptying after cholecystokinin stimulation was reduced in 69% of them. In 74% of patients at least one of the viscera under investigation showed abnormal motor function; however, only 36% of patients displayed involvement of the three organs. Gastrointestinal symptoms, duration and therapy of diabetes, previous poor glycemic control, and retinopathy did not correlate with the presence or the extent of motor disorders. Neuropathy was not predictive of gastrointestinal involvement and its extent; however, when motor abnormalities were present in patients with neuropathy, these were usually more severe. Gastrointestinal motor disorders are frequent and widespread in type 2 diabetics, regardless of symptoms. Autonomic neuropathy has a poor predictive value on motor disorders (0.75 for the esophagus, 0.5 for the stomach, 0.8 for the gallbladder), thus suggesting the coexistence of other pathophysiologic mechanisms. PMID- 10478881 TI - Helicobacter pylori antibody profile in household members of children with H. pylori infection. AB - Intrafamilial spread is implicated as a major route for acquisition of Helicoobacter pylori infection. Investigating H. pylori cytotoxin-associated protein (CagA) and vacuolating toxin (VacA) antibodies within family members enabled the authors to evaluate this possibility further. Serum samples were collected prospectively from household members after their index children were diagnosed with active H. pylori infection. Serum samples were evaluated for anti H. pylori immunoglobulin G antibody using the enzyme immunoassay (IEA) method and for H. pylori CagA and VacA antibodies with the commercially available immunoprobing Western blot kit. Ten different families participated in the study, including 10 pediatric patients and 31 household members. All patients and 28 household members (90%) were seropositive for H. pylori antibody by IEA and Western blot tests. Overall, 17 subjects (41.4%) were CagA positive, 14 (34.1%) were VacA positive, 11 (26.8%) were positive for both antibodies, and 22 (53.6%) were negative for both antibodies. A significant association in bacterial antibody profile was found between the patient index members and all household members (Cohen's kappa and Mentel-Haenszel methods). In four families, more than 66% of the household members harbored the same antibody profile, and in two families a completely different profile was observed. Moreover, a similar H. pylori antibody profile between the index patient and the mother was found in six families, and between the index patient and the father in two families. The data strongly suggest an intrafamiliar transmission for H. pylori infection. PMID- 10478882 TI - Does acid suppression by antacids and H2 receptor antagonists increase the incidence of atrophic gastritis in patients with or without H. pylori gastritis? AB - Currently there is controversial evidence that suggests that the accepted incidence of atrophic gastritis of 1.2 to 3.3% in patients with Helicobacter pylori gastritis may be increased by the long-term suppression of acid by a proton pump inhibitor (omeprazole). The purpose of this study is to show whether lesser forms of acid suppression by antacids or H2 receptor antagonists may have an influence on the development of atrophic gastritis. The authors recently reported a study in which a cohort of 36 patients with symptoms of dyspepsia were followed clinically for a period of 7 to 19 years. In that report all subjects underwent upper endoscopy with two biopsy specimens each from the antrum and fundus, on at least two occasions, 7 to 19 years apart. A diagnosis of atrophic gastritis was based on the interpretation of these biopsies by two gastrointestinal pathologists. The presence of H. pylori colonization was determined by tissue sampling and by a campylobacter-like organisms test of the antrum. Of the 36 patients in the authors' previous report, 33 had adequate baseline and follow-up data on medications consumed throughout the period of the study. In their current report they now present the findings of a retrospective review in which they correlate the presence of atrophic gastritis with the sole use of antacids and H2 receptor antagonists throughout the period of the study. In the cohort of 33 patients evaluated from the previous report, the authors found that atrophic gastritis had developed in all 28 patients positive for H. pylori, and in none of the 5 patients negative for H. pylori (p < 0.0001). A retrospective analysis of this previously studied cohort of 33 patients revealed that the use of antacids and H2 receptor antagonists did not predict the development of atrophic gastritis in either H. pylori-negative or -positive subjects. In a retrospective analysis of a cohort of 33 patients followed for an average of 11.7 years, atrophic gastritis developed in H. pylori-positive but not in H. pylori-negative subjects, irrespective of the use and duration of antacids or H2 receptor antagonists. PMID- 10478883 TI - Obstructive jaundice as the first clinical manifestation of a metastatic malignant melanoma in the ampulla of vater. AB - The authors report a patient with obstructive jaundice as the first clinical manifestation of a primary malignant melanoma metastatic in the ampulla of Vater. After the histologic diagnosis of the metastases was made, the primary tumor was located in the skin of the back. Obstruction jaundice secondary to ampullary obstruction due to metastatic melanoma has only been reported twice, and in neither case was jaundice the first symptom of the disease. PMID- 10478884 TI - Preservative treatment for biliobiliary fistula. AB - Biliobiliary fistula is thought to be a rare type of internal biliary fistula. A 68-year-old man presented to our hospital with complaints of jaundice and general malaise. Endoscopic retrograde cholangiography was performed, but the gallbladder was not imaged. Mirizzi's sign was observed in the common hepatic duct. During the course of evaluating this inpatient, imaging revealed that a gallstone was extruded to the right hepatic duct. After incision of the papilla, stones in the bile duct were subjected to mechanical lithotripsy and were extracted. As a result of bile duct decompression, the biliobiliary fistula was closed completely 2 months later. PMID- 10478885 TI - Enterolith ileus complicating jejunal diverticulosis. AB - Small intestinal obstruction caused by enteroliths originating in jejunal diverticula is rare. We present our experience with three elderly female patients who were successfully treated for this condition. The diagnosis, in all three cases, was made on laparotomy. Two patients underwent enterolithotomy while the third, who had a acutely inflamed jejunal diverticulum, had a jejunal resection. PMID- 10478886 TI - A Brunner's gland adenoma as a cause of anemia. AB - Benign tumors of the small bowel are rare. They present with many different manifestations depending on the size and location, and also cause a variety of symptoms that are often nonspecific. These include abdominal pain, dyspepsia, nausea, vomiting, and gastrointestinal bleeding that may be melena or hematemesis. Most of the time patients are asymptomatic and the lesions are discovered as an incidental finding. When bleeding occurs, and it may be severe in certain situations, the patient may develop signs of anemia, such as dyspnea, fatigue, and even high-output cardiac failure. The authors present a patient who was evaluated for melena and who was found to have a duodenal polyp that proved to be a Brunner's gland adenoma on pathology. PMID- 10478887 TI - Anorectal melanoma metastatic to the breast. AB - Anorectal melanoma is an extremely rare malignancy with poor prognosis. Patients generally present with a sensation of mass and rectal bleeding, which is usually attributed to hemorrhoids or polyps. It can not be diagnosed early because of these benign symptoms, so it is bulky at the time of presentation. Despite aggressive surgery, 5-year survival is less than 10%. We present a case of inoperable anorectal melanoma which metastasized to the left breast and abdominal lymph nodes. We also briefly reviewed the appropriate literature, emphasizing the diagnostic and therapeutic approaches. PMID- 10478888 TI - Rectal ulcers: a rare gastrointestinal manifestation of systemic lupus erythematosus. AB - A patient with systemic lupus erythematosus (SLE) developed a rectal ulcer and sepsis from colonic bacteria. At that time she had no other clinical manifestations of SLE. Histopathologic examination of the biopsies taken from the ulcer found evidence of vasculitis. Treatment with high-dose systemic steroids healed the ulcer clinically and endoscopically, but symptoms recurred when steroids were tapered. The patient was referred for surgery. This is a rare but dangerous complication of SLE and can be the only clinical manifestation of the disease. PMID- 10478889 TI - Long-term interferon alpha maintenance therapy for chronic hepatitis C infection in a patient with common variable immune deficiency. AB - A 46-year-old woman with common variable immune deficiency acquired acute non-A, non-B hepatitis from contaminated intravenous gamma globulin in 1983. For 6 years she had fluctuating elevations of her serum aminotransferase levels. In 1990 her serum was documented to be hepatitis C virusribonucleic acid positive by polymerase chain reaction, and her liver biopsy revealed chronic hepatitis with early cirrhosis (Knodell score, 15 points). Hepatitis C virus genotyping indicated that she had been infected with the type 3 genotype. She subsequently underwent treatment with interferon alpha (IFN-alpha) for 1 year and experienced biochemical, virologic, and histologic (Knodell score, 9) suppression. She was continued on maintenance therapy for an additional 7 years, with sustained biochemical and virologic suppression. During the sixth year of therapy, complications of portal hypertension were noted with mild ascites and eventually bleeding esophageal varices. This case report documents a favorable biochemical, virologic, and histologic response to IFN-alpha therapy in this setting; supports the notion that the natural progression of hepatitis C virus infection may be more aggressive in patients with common variable immune deficiency; and, although complications of portal hypertension eventually occurred, the suppressive maintenance IFN therapy may have delayed their onset. The future establishment of the long-term effects of IFN therapy on important clinical outcomes is necessary to understand better its therapeutic benefit in chronic hepatitis C infection. PMID- 10478890 TI - Development of hepatocellular carcinoma in a patient with chronic hepatitis C after 6 years of a sustained and complete response to IFN-alpha. AB - The authors report a rare case of hepatocellular carcinoma (HCC) that developed 6 years after a sustained and complete response to interferon (IFN) therapy for chronic hepatitis C. A 61-year-old Japanese man presented with a mass in the liver that was diagnosed as HCC. Six years earlier he was treated with IFN-alpha and responded successfully to therapy, with sustained normalization of serum aminotransferases and eradication of serum hepatitis C virus (HCV)-ribonucleic acid (RNA). HCV-RNA was also not detected in the resected tumorous and nontumorous liver tissues. The findings suggest that all patients with chronic HCV infection should be followed closely for as long as possible for the potential development of HCC even after a complete and sustained response to IFN treatment. PMID- 10478891 TI - Atrophy of the left lobe of the liver and anomalous hepatic vessel in a patient with Yunis-Varon syndrome. AB - Yunis-Varon syndrome is inherited as an autosomal recessive trait. It is characterized by facial and digit anomalies. This report describes a young woman with clinical features of this syndrome, atrophy of the left lobe of the liver, and a vascular anomaly. Liver abnormalities have not been described as features of this rare syndrome. PMID- 10478892 TI - Comments on current therapies for achalasia. PMID- 10478893 TI - The successful use of telemedicine in acute variceal hemorrhage. PMID- 10478894 TI - Intestinal occlusion and spigelian hernia. PMID- 10478895 TI - Sepsis due to Neisseria meningitidis manifested as acute cholecystitis. PMID- 10478896 TI - The economic value of treatment for depression. PMID- 10478897 TI - Acute psychopharmacological management of the aggressive psychotic patient. PMID- 10478898 TI - Teaching fundamental workplace skills to persons with serious mental illness. PMID- 10478899 TI - The diagnosis and treatment of depression in dementia. PMID- 10478900 TI - Introduction: preparing for the future. PMID- 10478901 TI - The epidemiology of common late-life mental disorders in the community: themes for the new century. AB - OBJECTIVE: The prevalence and incidence of the major mental disorders of late life that are common in the community and in primary health care are reviewed. METHODS: Community-based studies in English that included older adults were identified through MEDLINE searches and were reviewed. RESULTS: As the population ages, dementia, depression, and other mental conditions of the aged will demand more attention from clinicians and investigators to minimize their effects on disability, the use of health care services, and the quality of life for older adults and caregivers. Up to 15 to 20 percent of older adults have significant depressive symptoms, and it is estimated that as many as 45 percent of persons age 85 years and older have significant cognitive impairment and dementia. Other mental-health-related conditions, such as anxiety disorders, alcohol abuse, and prescription medicine misuse, are also important considerations but have not been as well studied as depression and dementia. Because an increasing proportion of older adults are members of minority groups, clinicians need to increase their awareness of how cultural factors relate to risk for mental disorders in late life. CONCLUSIONS: Attention to three themes may help clinicians and investigators meet the challenge of treating the common mental disorders of later life: the effect of these disorders on functioning, prevention of the consequences of mental disorders, and integration of mental health care and primary health care services. PMID- 10478902 TI - Depression and aging: a look to the future. AB - Geriatric depression is widespread, affecting at least one of six patients treated in general medical practice and an even higher percentage in hospitals and nursing homes. Depression in later life has serious consequences, including patients' and caregivers' distress, amplification of disability associated with medical and cognitive disorders of later life, increased health care costs, and increased mortality related to suicide and medical illness. Despite the numerous pathways to late-life depression, as well as the complex medical and psychosocial context in which it occurs, it can be effectively diagnosed and treated. To meet the public health challenges posed by geriatric depression over the next 15 years, the major priorities for research and clinical practice must include improvement in recognizing and treating depression among elderly patients seen in the general medical sector, suicide prevention among elderly patients seen in primary care, and acceleration of response to antidepressants. Other major priorities include improving the early recognition of treatment resistance in late-life depression and developing strategies for improving the treatment response of such patients; finding maintenance treatments with long-term efficacy, especially for patients over age 70 and those who have required electroconvulsive therapy; and developing preventive strategies to reduce the liability to late-onset depression. PMID- 10478903 TI - Alzheimer's disease and its management in the year 2010. AB - By 2010 the number of cases of Alzheimer's disease and other dementias will have increased by at least 25 percent. Alzheimer's disease poses an enormous threat to health service and public health resources. Three areas that require targeted research are early detection and recognition, biological markers and diagnosis, and pharmacotherapy. Care of patients with Alzheimer's disease in managed care organizations must be improved through a combination of research, education, advocacy, and legislation. Research in the pathogenic cascade of events within the brain leading to Alzheimer's disease has advanced, and treatment targets within the steps of the process have been specified. The current drug discovery and testing infrastructure is insufficient for major advances in therapy by 2010. Strategies for achieving an optimum outcome by then include increased funding of targeted research; expansion of drug discovery and therapeutic testing efforts; increased training of basic, clinical, and translational scientists; and study of optimum health care delivery systems. PMID- 10478904 TI - Older patients with schizophrenia: challenges in the coming decades. AB - The number and proportion of older adults with schizophrenia will increase considerably in the coming decades. Although a vast literature on schizophrenia among younger adults exists, much less is known about late-life schizophrenia and its treatment. The authors describe two potential scenarios for 2011, the year that the first baby boomers will turn 65. To ensure that the more favorable scenario becomes a reality, the authors suggest four goals: decrease medical comorbidity and mortality among younger patients with schizophrenia and improve their access to health care so that they can live longer and more productive lives; improve our understanding of the neurobiological and psychosocial factors underlying late-life schizophrenia, as well as the health care and social service needs of such patients; develop more effective and safer pharmacologic, psychosocial, and cognitive behavioral treatments; and improve rehabilitation of older people with schizophrenia. Specific strategies to foster these goals include establishing a consortium for studies of late-life schizophrenia; conducting multicenter studies of treatment effectiveness; and forming interdisciplinary collaborations among researchers, clinicians, government and industry representatives, and patient advocacy groups. PMID- 10478905 TI - The potential impact of the baby-boom generation on substance abuse among elderly persons. AB - Little attention has been paid to substance use disorders in the elderly population. Currently available diagnostic criteria are likely to significantly underestimate the prevalence of substance abuse among elderly persons because they were developed and validated in younger samples. As baby boomers age, the number of elderly persons who misuse or abuse illicit drugs and alcohol may increase because this age cohort has higher rates of use of these substances than previous cohorts. Abuse and misuse of prescription and over-the-counter drugs may also increase due to the larger numbers of baby boomers. Few studies have addressed treatment issues that may be unique to elderly substance abuse patients. Some evidence suggests that substance abuse treatment outcomes are poorer among individuals with cognitive impairment, and special treatment strategies are needed for elderly persons with dementia. To identify the magnitude of the problem, diagnostic criteria should be modified and national survey data should be analyzed to provide more accurate estimates of substance abuse and dependence among baby boomers. PMID- 10478906 TI - Community-based long-term care for older persons with severe and persistent mental illness in an era of managed care. AB - The authors describe current needs and trends in the mental health care, including long-term care, of older persons with severe and persistent mental illness. The literature suggests that emerging models of managed long-term care hold promise for integrated services but do not currently address the specialized mental health needs of this patient group. The authors review issues in financing long-term mental health care, including controversies over fee-for-service and carve-out and carve-in arrangements. Without mechanisms to adequately finance services, adjust for risk, and measure outcomes, the authors conclude, managed care arrangements will be in conflict with the goal of high-quality care for older adults with severe and persistent mental illness. Proposed directions for future models of care for this group include integration of mental health and medical services, integration of specialized geropsychiatric services with developing community-based long-term care systems, blended financing under shared risk arrangements, and assurance of accountability and outcomes under managed care. PMID- 10478907 TI - The future of psychosocial treatments for elderly patients. AB - Geriatric psychiatric disorders usually occur in the context of medical illness, disability, and psychosocial impoverishment. Preliminary evidence suggests that psychotherapy can reduce not only psychopathology but also physical complaints, pain, and disability and that it can improve compliance with medical regimens. Psychotherapy has been found effective in treatment of depression related to bereavement and caregiver burden. Modification of available treatment strategies to address infirmity and life adversity may have a major impact on rehabilitation from psychiatric and medical disorders and may reduce utilization of nonpsychiatric health services. Most elderly patients with psychiatric problems prefer to be treated by their primary care physician. Models need to be developed and tested to integrate psychotherapy and other mental health services in primary care settings so that timely and appropriately targeted interventions can be provided. PMID- 10478908 TI - Training in geriatric mental health: needs and strategies. AB - This paper examines current numbers of U.S. mental health care professionals in psychiatry, psychology, social work, and nursing who are trained to treat late life mental disorders and presents estimates of the increased numbers that will be needed by 2010. Strategies that may be effective in increasing the supply of geriatric mental health specialists are expanding incentives, such as fellowship stipends, loan forgiveness programs, and clinical reimbursement, to attract more clinicians to the field; encouraging professional organizations to offer certification in aging to highlight increased knowledge among professionals; and increasing core curricula in late-life mental disorders, or at least in aging, in all health professions. Other strategies are to diversify the range of training opportunities available by establishing short-term training initiatives to meet specific demands; use new technology to provide education to both professionals and the public; and establish "centers of excellence" to advance research training and clinical care in late-life mental disorders. PMID- 10478909 TI - A community study of depression treatment and employment earnings. AB - OBJECTIVE: Although treatment for major depression has been shown to reduce the costs of lost earnings resulting from lost work days, research has not demonstrated whether the reduction fully offsets the costs of treatment for the disorder. METHODS: A statewide cohort of community residents with recent major depression, dysthymia, or substantial depressive symptoms was recruited and interviewed at baseline and at six-month and 12-month follow-ups. The cost of lost earnings was measured by lost work days multiplied by subjects' wage rates. Cost of treatment for depression was approximated using charges abstracted from provider and insurance records. Net economic cost, defined as the sum of changes in lost earnings and depression treatment costs, was examined in multiple regression analyses. RESULTS: After the analyses controlled for sociodemographic characteristics, baseline severity of depression, and comorbidity, no statistically significant effect of depression treatment on net economic cost was found. This finding suggests that the cost of depression treatment was fully offset by savings from reduction in lost work days. Results from sensitivity analyses in multiple alternative scenarios support the same conclusion. CONCLUSIONS: The finding of a full offset of depression treatment cost is conservative because other benefits, such as reduced pain and suffering and increased productivity while at work, were not included in the analyses. Employers who bear the cost from lost work days should encourage their employees with depressive disorders to seek treatment, even if it means paying for the entire treatment cost. Self-employed individuals with depression also will benefit even if they pay for the treatment costs themselves. PMID- 10478910 TI - Siblings of adults with mental illness or mental retardation: current involvement and expectation of future caregiving. AB - OBJECTIVE: The study examined the factors associated with the involvement of siblings in the life of a brother or sister who has mental illness or mental retardation. Involvement was defined as the current provision of instrumental and emotional support as well as the expectation of future caregiving responsibility. METHODS: A mailed questionnaire was used to collect data from 61 siblings of adults with serious mental illness and 119 siblings of adults with mental retardation. The sample was drawn from two ongoing longitudinal studies. RESULTS: The two groups of siblings showed striking differences in their expectations about their responsibility for future caregiving. Almost 60 percent of the siblings of adults with mental retardation expected to assume primary caregiving responsibility in the future, but only one-third of the siblings of adults with mental illness held this expectation. For both groups, competing family responsibilities limited the involvement of siblings, whereas closeness to the family of origin led to greater sibling involvement. CONCLUSIONS: The extent of current and future involvement by siblings of adults with disabilities is a function of the demands and constraints of midlife as well as the degree of closeness with the family of origin. The findings highlight the importance of clinicians' work to support and strengthen family relationships, which loom large in determining the extent to which siblings are involved in the care of a brother or sister with disabilities. PMID- 10478911 TI - The Massachusetts forensic transition program for mentally ill offenders re entering the community. AB - On April 1, 1998, the Massachusetts Department of Mental Health established the forensic transition program for mentally ill offenders. The primary goal of the statewide program, which follows clients for three months after their release from correctional facilities, is to coordinate services and assist in community reintegration. In the first year of the program, 233 mentally ill offenders received services. Seventy-four clients had been discharged as of April 1, 1999. At discharge 42 of the 74 clients (57 percent) were living in the community and were receiving mental health services. The other clients were hospitalized immediately after release (20 percent), reincarcerated (10 percent), hospitalized after a brief stay in the community (3 percent), or lost to follow-up (11 percent). PMID- 10478912 TI - Patterns of family caregiving and support provided to older psychiatric patients in long-term care. AB - Data on patterns of relationships and caregiving between older, institutionalized chronically mentally ill patients and their families were gathered in brief face to-face interviews with 109 patients randomly selected from residents age 45 or older in a large intermediate care facility in Chicago. Three-fourths of the sample maintained some form of family contact. One-third had been married or had children. Siblings were the most frequently identified family contact and support. The results suggests that older, institutionalized psychiatric patients continue to have family contact and that siblings and offspring become increasingly important as patients age. PMID- 10478913 TI - AIDS-related services and training in outpatient mental health care agencies in New York. AB - Directors of 471 outpatient mental health settings in New York State (82.1 percent of 574 settings located in counties with intermediate to high AIDS case rates) completed a survey about HIV and AIDS services, training needs, and barriers to care. Most of the sites served one to ten persons with HIV infection annually and had staff members who were trained in providing at least one HIV related service. Nonetheless, 84 percent of the respondents reported unmet needs for training. The likelihood of providing certain services was significantly increased in sites that were in urban locations, primarily served clients with comorbid alcohol or other drug use disorders, lacked funds for providing condoms, had staff members who were trained in HIV and AIDS services, identified particular HIV training needs, believed clients needed condoms, and viewed HIV related services as very important. PMID- 10478914 TI - Initiatives for improving mental health services to ethnic minorities in Australia. AB - Several major policy initiatives have been undertaken in Victoria, Australia's second-largest state, to address difficulties in providing accessible and culturally sensitive mental health services to ethnic minorities. These initiatives include the development and publication of a policy statement, alteration of funding formulas to take into account populations of persons with non-English-speaking backgrounds, incentive funding mechanisms, new funding specifically devoted to ethnic mental health, the establishment of service development positions in the major public hospital networks, and the establishment of a pilot project to employ bilingual staff as psychiatric case managers. PMID- 10478915 TI - Advance directives. PMID- 10478916 TI - Advance directives. PMID- 10478917 TI - Medication in depression. PMID- 10478918 TI - The doctor-patient relationship. PMID- 10478919 TI - More than a quarter million inmates in U.S. prisons and jails are mentally ill, Justice Department report finds. PMID- 10478920 TI - Proposed federal rules will shift oversight, increase accessibility of methadone maintenance treatment. PMID- 10478921 TI - Surgeon General's call to action on suicide stresses strategies for prevention. PMID- 10478922 TI - Cloning of delta12- and delta6-desaturases from Mortierella alpina and recombinant production of gamma-linolenic acid in Saccharomyces cerevisiae. AB - Two cDNA clones with homology to known desaturase genes were isolated from the fungus Mortierella alpina. The open reading frame in one clone encoded 399 amino acids and exhibited delta12-desaturase activity when expressed in Saccharomyces cerevisiae in the presence of endogenous fatty acid substrate oleic acid. The insert in another clone contained an open reading frame encoding 457 amino acids and exhibited delta6-desaturase activity in S. cerevisiae in the presence of exogenous fatty acid substrate linoleic acid. Expression of the delta12 desaturase gene under appropriate media and temperature conditions led to the production of linoleic acid at levels up to 25% of the total fatty acids in yeast. When linoleic acid was provided as an exogenous substrate to the yeast cultures expressing the delta6-desaturase activity, the level of gamma-linolenic acid reached 10% of the total yeast fatty acids. Co-expression of both the delta6 and delta12-desaturase cDNA resulted in the endogenous production of gamma linolenic acid. The yields of gamma-linolenic acid reached as high as 8% of total fatty acids in yeast. PMID- 10478923 TI - Intermediates and products formed during fatty acid alpha-oxidation in cucumber (Cucumis sativus). AB - Fatty acid alpha-oxidation is an essential metabolic pathway both in plants and in mammals which is still not completely understood. We previously described and purified an alpha-oxidation enzyme in cucumber which has been used in the present investigation of the alpha-oxidation reaction mechanism. Free fatty acids, and not the CoA thioesters, were found to undergo alpha-oxidation in cucumber. 2 Hydroxy- and 2-oxopalmitic acids were identified as palmitic acid alpha-oxidation intermediates by high-performance liquid chromatography and gas chromatography mass spectrometry analysis in cucumber subcellular 150,000 x g(max) pellets obtained by differential centrifugation. Incubation of purified alpha-oxidation enzyme with [1-14C]palmitic acid resulted in the formation of both the above described intermediates and the Cn-1 product, pentadecanal, and 14CO2. Besides 14CO2, 14C-formate was identified as an alpha-oxidation product from [1 14C]palmitic acid in cucumber subcellular fractions. Fe2+ stimulated the 14CO2 and 14C-formate production, and the addition of ascorbate and 2-oxoglutarate together with Fe2+ resulted in optimal alpha-oxidation activities, suggesting a dioxygenase reaction mechanism, as previously shown in mammals. NADPH and, to a lesser extent, NADH stimulated the total 14C-formate plus 14CO2 production but had only slight or no effects on 14CO2 production. H2O2 showed concentration dependent inhibitory effects, while FAD had neither effect on 14CO2 nor 14CO2 plus 14C-formate production. The results in the present study demonstrate that an alpha-oxidation enzyme in cucumber is capable of oxidizing palmitic acid via 2 hydroxy- and 2-oxo-palmitic acid to produce pentadecanal and CO2. In contrast to the subcellular 150,000 x g(max) fraction, the purified alpha-oxidation enzyme could neither produce formate nor convert 14C-formate into 14C02, indicating two possible alpha-oxidation routes in cucumber. PMID- 10478924 TI - Proportion of geometrical hydroperoxide isomers generated by radical oxidation of methyl linoleate in homogeneous solution and in aqueous emulsion. AB - The proportion of geometrical hydroperoxide isomers generated by aerobic oxidation of methyl linoleate (18:2 Me) in either aqueous emulsion consisting of Tris-HCl buffer (pH 7.4) or in a homogeneous dichloromethane solution was determined to understand the mechanism of lipid oxidation in different reaction systems. Four geometrical isomers were generated after oxidation of 18:2 Me in dichloromethane: methyl 13-hydroperoxy-cis-9, trans-11-octadecadienoate, methyl 13-hydroperoxy-trans-9,trans-11-octadecadienoate, methyl 9-hydroperoxy-trans 10,cis-12-octadecadienoate, and methyl 9-hydroperoxy-trans-10, trans-12 octadecadienoate in the ratios of 1:4:1:4, respectively. The ratios between each isomer did not change until the peroxide value (PV) increased to 58 meq/kg. Oxidation of 18:2 Me in aqueous emulsion yielded the same geometrical isomers of hydroperoxide. However, the ratios were different: 3:2:3:2 until the PV increased to 110 meq/kg. Predominant (60%) formation of trans,trans hydroperoxide isomers was obtained in the oxidation of a mixture of 18:2 Me and methyl laurate (12:0 Me). These results are interpreted to reflect the importance of the concentration of hydrogen atom-donating equivalents to the kinetic preference for different products. The high effective concentration of hydrogen donors in the oxidation of 18:2 Me in emulsions favored the formation of the less stable cis,trans isomers. The lower concentration of hydrogen donor in the dichloromethane solution effectively slowed hydrogen donation and led to the strong preference for the more stable trans,trans isomers. This interpretation was further tested by preparing emulsions of 18:2 Me and 12:0 Me to dilute concentration of hydrogen donating species using the nonhydrogen-donating 12:0 Me. Consistent with the proposed hypothesis, the proportion of trans,trans isomers increased as a result of 12:0 Me addition. PMID- 10478925 TI - 19-Azasqualene-2,3-epoxide and its N-oxide: metabolic fate and inhibitory effect on sterol biosynthesis in Saccharomyces cerevisiae. AB - 19-Azasqualene-2,3-epoxide was more inhibitory than the corresponding N-oxide against 2,3-oxidosqualene cyclase (OSC) solubilized from Saccharomyces cerevisiae (IC50 7+/-2 and 25+/-5 microM, respectively). Both compounds showed a reversible, noncompetitive-type inhibition on solubilized OSC. Different inhibitory properties between the compounds were especially evident when measuring [14C]acetate incorporation into nonsaponifiable lipids extracted from treated cells. In cells treated with 19-azasqualene-2,3-epoxide at 30 microM, the radioactivity associated with the oxidosqualene fraction, which was negligible in the controls, rose to over 40% of the nonsaponifiable lipids, whereas it remained at a slightly appreciable level in cells treated with the N-oxide derivative under the same conditions. 19-Azasqualene-2,3-epoxide was also more effective than the N-oxide as a cell growth inhibitor (minimal concentration of compound needed to inhibit yeast growth: 45 and >100 microM, respectively). The two inhibitors underwent different metabolic fates in the yeast: while 19-azasqualene 2,3-epoxide did not undergo any transformation, its N-oxide was actively reduced to the corresponding amine in whole and in "ultrasonically stimulated" cells. The N-oxide reductases responsible for this transformation appear to be largely confined within the microsomal fractions and require NADPH for their activity. A possible relationship between the inhibitory properties of the two compounds and their metabolic fates is discussed. PMID- 10478926 TI - C2-ceramide attenuates phenylephrine-induced vasoconstriction and elevation in [Ca2+]i in rat aortic smooth muscle. AB - In the present study, we examined the effects of cell-permeable C2-ceramide on contraction of aortic smooth muscle and intracellular free Ca2+ ([Ca2+]i). C2 ceramide (10(-7) to 10(-4) M) alone did not elicit any significant changes in either basal tension or resting levels of [Ca2+]i in rat aortic smooth muscle. However, C2-ceramide (10(-7) to 10(-4) M) attenuated phenylephrine-induced contractions in isolated rat aortic rings in a concentration-related manner, and inhibited elevations in [Ca2+]i in cultured rat aortic smooth muscle cells induced by phenylephrine. C2-ceramide-induced relaxation was found to be only slightly endothelium-dependent. However, nitric oxide inhibitors (L-NNA, L-NMMA), an inhibitor of prostanoid synthesis (indomethacin), an inhibitor of opiate actions, and several inhibitors of the pharmacologic actions of various vasoactive amines all failed to interfere with the vasorelaxant responses of C2 ceramide. Three different inhibitors of protein kinase C, when used in a wide concentration range, also failed to interfere with the ceramide-induced relaxations. Our results suggest that the sphingomyelin-signaling pathway may play an important regulatory role in arterial wall tone. PMID- 10478927 TI - Specific modifications of phosphatidylinositol and nonesterified fatty acid fractions in cultured porcine cardiomyocytes supplemented with n-3 polyunsaturated fatty acids. AB - Mechanisms for the antiarrhythmic effect of n-3 polyunsaturated fatty acids (PUFA) are currently being investigated using isolated cardiac myocytes. It is still not known whether the incorporation of n-3 PUFA into membrane phospholipids is a prerequisite for its protective action or if n-3 PUFA exert antiarrhythmic effects in their nonesterified form as demonstrated by recent studies. Adult porcine cardiomyocytes were grown in media supplemented with arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). After 24 h, analysis of total lipids showed that the myocytes were enriched with the respective fatty acids compared to control cells. Large proportions of all three fatty acids supplemented (69% AA, 72% DHA, and 66% EPA) remained unesterified. Fatty acid analyses of total phospholipids (PL) revealed that the incorporation of EPA and DHA, though small, was significantly different (P<0.05) from that of the control cells. The PL fraction was further separated into phosphatidylinositol (PI), phosphatidylethanolamine, phosphatidylcholine, and phosphatidylserine to study the pattern of incorporation of the fatty acids in these fractions. It became apparent that EPA and DHA were selectively incorporated into the PI fraction. This study demonstrates that in adult porcine cardiomyocytes, the n-3 PUFA supplementation selectively modulates two important lipid fractions, nonesterified fatty acid and PI, which were implicated in the mechanisms of prevention of cardiac arrhythmias. PMID- 10478928 TI - Dose-dependent inhibition of cell proliferation induced by lipid peroxidation products in rat hepatoma cells after enrichment with arachidonic acid. AB - Polyunsaturated fatty acids (PUFA) are important constituents of membrane phospholipids, whose levels are decreased in some tumor cells. This deficiency may cause alterations in signal transduction and an interruption of normal cellular events. The enrichment of tumor cells with PUFA may stimulate or inhibit tumor growth, probably depending on the type of PUFA and the cellular concentration of aldehydes derived from restored lipid peroxidation. We examined the effect of several doses of prooxidant on the growth of hepatoma cells with different aldehyde dehydrogenase activities, enriched with arachidonic acid. Two doses of prooxidant were sufficient to reduce growth of hepatoma cells with low aldehyde dehydrogenase activity, whereas three doses were necessary for those with high enzyme activity. In both cases, lipid peroxidation products blocked the cells in the S phase. PMID- 10478929 TI - Ethanol-extracted soy protein isolate results in elevation of serum cholesterol in exogenously hypercholesterolemic rats. AB - Soy protein preparations were reported to have hypocholesterolemic actions in experimental animals and humans, while the active components and the mechanism by which this occurs are not clarified yet. The objective of this study is to address these issues by using exogenously hypercholesterolemic rats which are susceptible to dietary cholesterol. Two groups of five rats (male, 12-wk-old) were fed on AIN 93G-based diet with soy protein isolate (SPI) or ethanol extracted SPI (EE-SPI) for 2 wk. EE-SPI was prepared by ethanol extraction to remove isoflavones and other components. Concentrations of serum and liver total cholesterol were lower in rats fed SPI than in those fed EE-SPI. The abundances of mRNA for 7alpha-hydroxylase and low density lipoprotein receptor in the liver were lower in EE-SPI group than those in SPI group. These results suggest that the ethanol extract from SPI has a factor(s) to alleviate hypercholesterolemia by increasing the removal of cholesterol from serum through the receptor pathway and then from liver through enhancement of bile acid synthesis. PMID- 10478930 TI - The effects of a dietary oxidized oil on lipid metabolism in rats. AB - This study was carried out to investigate the effects of a dietary oxidized oil on lipid metabolism in rats, particularly the desaturation of fatty acids. Two groups of rats were fed initially for a period of 35 d diets containing 10% of either fresh oil or thermally treated oil (150 degrees C, 6 d). The dietary fats used were markedly different for lipid peroxidation products (peroxide value: 94.5 vs. 3.1 meq O2/kg; thiobarbituric acid-reactive substances: 230 vs. 7 micromol/kg) but were equalized for their fatty acid composition by using different mixtures of lard and safflower oil and for tocopherol concentrations by individual supplementation with DL-alpha-tocopherol acetate. In the second period which lasted 16 d, the same diets were supplemented with 10% linseed oil to study the effect of the oxidized oil on the desaturation of alpha-linolenic acid. During the whole period, all the rats were fed identical quantities of diet by a restrictive feeding system in order to avoid a reduced food intake in the rats fed the oxidized oil. Body weight gains and food conversion rates were only slightly lower in the rats fed the oxidized oil compared to the rats fed the fresh oil. Hence, the effects of lipid peroxidation products could be studied without a distortion by a marked reduced food intake and growth. To assess the rate of fatty acid desaturation, the fatty acid composition of liver and heart total lipids and phospholipids was determined and ratios between product and precursor of individual desaturation reactions were calculated. Rats fed the oxidized oil had reduced ratios of 20:4n-6/18:2n-6, 20:5n-3/18:3n-3, 20:4n 6/20:3n-6, and 22:6n-3/22:5n-3 in liver phospholipids and reduced ratios of 20:4n 6/18:2n-6, 22:5n-3/18:3n-3, and 22:6n-3/18:3n-3 in heart phospholipids. Those results suggest a reduced rate of desaturation of linoleic acid and alpha linolenic acid by microsomal delta4-, delta5-, and delta6-desaturases. Furthermore, liver total lipids of rats fed the oxidized oil exhibited a reduced ratio between total monounsaturated fatty acids and total saturated fatty acids, suggesting a reduced delta9-desaturation. Besides those effects, the study observed a slightly increased liver weight, markedly reduced tocopherol concentrations in liver and plasma, reduced lipid concentrations in plasma, and an increased ratio between phospholipids and cholesterol in the liver. Thus, the study demonstrates that feeding an oxidized oil causes several alterations of lipid and fatty acid metabolism which might be of great physiologic relevance. PMID- 10478931 TI - Absorption in rats of rapeseed, soybean, and sunflower oils before and following moderate heating. AB - Rapeseed, soybean, and sunflower oil were heated for 15 min in a 5-mm oil layer in a pan at 180 degrees C. The fatty acid composition was almost unaffected by heating, while the polymer content rose slightly and the tocopherol content decreased, except in soybean oil. The absorption of oils before and after heating was investigated in lymph-cannulated rats. Oils were administered as emulsions through a gastrostomy tube and lymph was collected during the next 24 h. The highest accumulated lymphatic transport of total fatty acids was observed after administration of rapeseed oil, and the lowest after heated sunflower oil. The accumulated transport was similar for all unheated oils. The transport of fatty acids was significantly lower in rats receiving heated oil compared to those receiving the corresponding unheated oil. Small increases in polymers may have contributed to the decreased lymphatic transport of oil following heating, although this probably does not fully explain the effect. The absorption of sunflower oil was more affected by heating than the absorption of soybean or rapeseed oil. Furthermore, the largest decrease in total activity of tocopherols following heating was observed in sunflower oil. Overall, these results demonstrate that the absorption of vegetable oils is affected by moderate heating. PMID- 10478932 TI - Fatty acid composition of brain glycerophospholipids in peroxisomal disorders. AB - This paper shows for the first time the differential fatty acid composition of ethanolamine plasmalogens (EP) and phosphatidylethanolamine (PE) in the brains of 12 patients with disorders of peroxisomal biogenesis and compares the results to normal values for the age. Other important glycerophospholipids (GPL), such as phosphatidylserine (PS) and phosphatidylcholine (PC), are also included in this study. GPL were separated by two-dimensional thin-layer chromatography, and their fatty acid composition was determined by capillary column gas-liquid chromatography. Total brain GPL were slightly decreased in peroxisomal disorders (27.98+/-2.95 micromol/g in the patients against 34.5+/-6.21 micromol/g in age matched controls, P = 0.005), and the distribution of the different GPL classes was much altered. In confirmation of known data, EP were very much decreased (2.18+/-1.3 micromol/g in the patients against 6.9+/-2.3 micromol/g in controls) at the expense of PE, which was increased (8.58+/-2.17 micromol/g in the patients against 5.97+/-0.58 micromol/g in controls, P<0.005). PS and PC were both significantly decreased (P = 0.0001 and P = 0.037, respectively). The polyunsaturated fatty acid (PUFA) composition of all the GPL fractions was markedly abnormal. In absolute terms, docosahexaenoic acid (22:6n-3) was drastically decreased in all GPL classes (always at the P<0.0001 level) while arachidonic acid (20:4n-6) was increased in PE and PS (P<0.001 in both cases). In the alkenyl acyl form, EP, 22:6n-3, and 20:4n-6 were both very significantly decreased (P<0.0001), although the former was always the most affected. The myelin PUFA adrenic acid (22:4n-6) was decreased in EP (P<0.0001) and slightly increased in PS (P<0.05). The changes found confirm that 22:6n-3 deficiency is a predominant defect in the brain in peroxisomal disorders. PMID- 10478933 TI - Application of high-resolution, two-dimensional 1H and 13C nuclear magnetic resonance techniques to the characterization of lipid oxidation products in autoxidized linoleoyl/linolenoylglycerols. AB - Subjection of polyunsaturated fatty acid (PUFA)-rich culinary oils to standard frying episodes generates a range of lipid oxidation products (LOP), including saturated and alpha,beta-unsaturated aldehydes which arise from the thermally induced fragmentation of conjugated hydroperoxydiene precursors. Since such LOP are damaging to human health, we have employed high-resolution, two-dimensional 1H-1H relayed coherence transfer, 1H-1H total correlation, 1H-13C heteronuclear multiple quantum correlation, and 1H-1H J-resolved nuclear magnetic resonance (NMR) spectroscopic techniques to further elucidate the molecular structures of these components present in (i) a model linoleoylglycerol compound (1,3 dilinolein) allowed to autoxidize at ambient temperature and (ii) PUFA-rich culinary oils subjected to repeated frying episodes. The above techniques readily facilitate the resolution of selected vinylic and aldehydic resonances of LOP which appear as complex overlapping patterns in conventional one-dimensional spectra, particularly when employed in combination with solvent-induced spectral shift modifications. Hence, much useful multi-component information regarding the identity and/or classification of glycerol-bound conjugated hydroperoxydiene and hydroxydiene adducts, and saturated and alpha,beta-unsaturated aldehydes, present in autoxidized PUFA matrices is provided by these NMR methods. Such molecular information is of much value to researchers investigating the deleterious health effects of LOP available in the diet. PMID- 10478934 TI - Recognition and quantitation of cis-vaccenic and eicosenoic fatty acids in olive oils by 13C nuclear magnetic resonance spectroscopy. AB - The presence of 11-cis monoenoic fatty acids was detected in olive oil samples by means of 13C nuclear magnetic resonance spectroscopy, and the positional isomery on the glycerol backbone was derived. The 11-cis vaccenic and eicosenoic fatty acid resonances were recognized and the amounts of the fatty acids quantified. For comparison purposes, a quantitative analysis was also made by gas chromatography. PMID- 10478935 TI - Effect of dietary lipids on phospholipase D activity in rat brain. AB - Phospholipase D (PLD) is emerging as a major player in many novel signaling pathways. Based on recent studies correlating membrane composition with enzyme function, we speculated that feeding of dietary lipids to the newborns has a major impact on brain PLD activity. To test this hypothesis, the rat dams were fed fat-free powder containing either safflower oil or fish oil, and a control powdered chow. The pups were weaned onto the diet and sacrificed at 30 days of age. PLD activity was measured by transphosphatidylation assays using rat brain membranes. This study shows that microsome GTPgammaS-dependent PLD activity in rats fed safflower oil or fish oil was significantly reduced by 38% and 30% respectively compared to controls. Oleate-dependent PLD activity in the safflower oil group, however, was significantly increased by 38%. In contrast, synaptosome membrane (P2) GTPgammaS-dependent PLD activity in rats consuming safflower oil was significantly increased by 29%, but there was no difference in oleate dependent PLD activity. Likewise, no difference was observed in microsome oleate dependent PLD and P2 GTPgammaS-dependent PLD activity between the fish oil and the control groups. These results indicate that dietary lipid intake appears to modulate phospholipid metabolism and differential expression of PLD isozymes in the brain. PMID- 10478936 TI - Stat3 participates in the metabotropic glutamate signaling pathway in Bergmann glial cells. AB - Purkinje cells establish a glutamatergic synapse with the parallel fibers in the cerebellum. Bergmann glial cells surround this synapse. The transcription factor Stat3, highly expressed in cerebellum, is present in cultures of chick Bergmann glia. Treatment of these cells with L-glutamate induces the tyrosine phosphorylation of Stat3. The maximal phosphorylation was observed after 15 min and by 30 min returned to basal levels. This effect correlated with Stat3 translocation to nucleus and the appearance of a retarded band in gel mobility shift assays. The effects observed with L-glutamate were mediated through metabotropic glutamate receptors. These results further support the notion that glial cells respond to L-glutamate through changes in the existing proteins and also in gene expression regulation. PMID- 10478937 TI - Enzyme activity and protein of multiple forms of choline acetyltransferase: effects of calyculin A and okadaic acid. AB - Choline acetyltransferase (ChAT) appears to exist in multiple forms, three of which can be isolated biochemically as cytosolic (cChAT), ionically-membrane bound (ibChAT) and non-ionic membranous (mChAT). In this study, we first examined whether the quantitative distribution of enzyme protein and enzyme activity was the same. Enzyme activity and ChAT protein distributed similarly: the majority of ChAT activity and protein were found in cChAT followed by mChAT and least activity and amount were in ibChAT. Our second objective was to investigate the effects of calyculin A or okadaic acid on the subcellular distribution of ChAT activity and amount from rat hippocampal formation. Calyculin A and okadaic acid decreased significantly (p < 0.01) cytosolic and membranous ChAT activity; ionically-bound ChAT was not significantly (p > 0.67) different from control. Removal of calyculin A or okadaic acid restored cytosolic ChAT activity (p > 0.9 as compared to control), but not membranous enzyme activity (p < 0.05 as compared to control). The immunoreactive cytosolic ChAT was reduced significantly (p < 0.01) by calyculin A and okadaic acid. Enzyme amount of membranous ChAT was decreased significantly by calyculin A (p < 0.01) and okadaic acid (p < 0.001). Enzyme amount of ionically-bound ChAT was not changed (p > 0.99) by either of these two phosphatase inhibitors. This investigation demonstrates that alterations in ChAT activity of each subfraction parallel changes in enzyme amounts in the same fractions. PMID- 10478938 TI - UDP-glucuronosyltransferase in the rat olfactory bulb: identification of the UGT1A6 isoform and age-related changes in 1-naphthol glucuronidation. AB - Xenobiotic glucuronidation represents a major metabolic protection of the brain against chemical aggressions at blood-brain interfaces. We previously observed that glucuronidation of 1-naphthol was very effective in olfactory bulb, which is a pathway for the entry of foreign molecules into the brain. In this work, we showed that 1-naphthol glucuronidation varied according to age. It was very high at birth, then decreased markedly in 3-month-old rats and increased again significantly during aging. By Western blot and reverse transcription-polymerase chain reaction (RT-PCR), we demonstrated the presence in the olfactory bulb of the UDP-glucuronosyltransferase (UGT) 1A6 isoform, which catalyzes the glucuronidation of phenols, such as 1-naphthol. Quantitative RT-PCR indicated that the mRNA levels encoding UGT1A6 did not significantly change according to age, thus suggesting that other differently regulated UGT isoforms were present and would account for the variations of 1-naphthol glucuronidation observed. PMID- 10478939 TI - Aluminum enhances melanin-induced lipid peroxidation. AB - The present study was conducted to characterize the possible interaction of Al3+ and Fe2+ with synthetic melanin in the potentiation of lipid peroxidation in liposomes and rat caudate-putamen homogenates. Al3+ stimulated melanin-initiated lipid peroxidation as measured by the production of 2-thiobarbituric acid reactive substances (TBARS) and conjugated dienes. The effect of A13+ was dependent on melanin (10-100 microg/ml) and A13+ (2.5-250 microM) concentrations and no synergism between Fe2+ and Al3+ was observed. The prooxidant effect of Al3+ was partially inhibited by superoxide dismutase indicating the involvement of O2*- . Ga3+ and Be2+ which can increase NADH oxidation in the presence of O2* , also were shown to stimulate melanin-initiated TBARS production. Based on the effect of Al3+ and other non redox metals, we suggest that Al3+ does not act through either the induction of melanin free radicals, or the induction of changes in membrane physical properties. Results show that Al3+ enhances melanin initiated lipid peroxidation in part through an interaction with O2*- generated from the autoxidation of melanin. We speculate that Al3+ contributes to neuromelanin-mediated oxidative damage in dopaminergic neurons and subsequent neuronal degeneration and death in Parkinson's disease. PMID- 10478940 TI - Mechanisms of D-aspartate release under ischemic conditions in mouse hippocampal slices. AB - The release of preloaded D-[3H]aspartate, an unmetabolizable analogue of L glutamate, was studied in superfused hippocampal slices from 7-day-old and 3 month-old (adult) mice under various cell-damaging conditions, including hypoxia, hypoglycemia, ischemia, oxidative stress and the presence of free radicals and metabolic poisons. The release was generally markedly enhanced in most of the above conditions, the responses being greater in adults than in developing mice. The presence of dinitrophenol had the most pronounced effect at both ages, followed by NaCN- and free-radical-containing media and ischemia. Hypoxia did not affect release in the immature hippocampus. Under most conditions K+ stimulation (50 mM) was still able markedly to enhance D-aspartate release. This potentiation under cell-damaging conditions in both adult and developing hippocampus signifies that increased L-glutamate release contributes to excitotoxicity and subsequent cell death. The mechanisms of ischemia-induced release of D-aspartate were analyzed in the adult hippocampus using ion channel inhibitors and modified superfusion media. The induced release proved to be partly Ca(2+)-dependent and partly Ca(2+)-independent. The results obtained with Na+ omission and homo- and heteroexchange with D-aspartate and L-glutamate demonstrated that a part of the release in normoxia and ischemia is mediated by the reversal of Na(+)-dependent glutamate transporters. The Na+ channel blockers amiloride and riluzole reduced the ischemia-induced release, also indicating the involvement of Na+ channels. In addition to this, the enhanced release of D-aspartate may comprise a swelling induced component through chloride channels. PMID- 10478941 TI - Reversible activation of glutamate transport in rat brain glia by protein kinase C and an okadaic acid-sensitive phosphoprotein phosphatase. AB - High-affinity L-glutamate (GLU) transport is an important regulator of excitatory amino acid (EAA) concentrations in brain extracellular fluid and may play a key role in excitatory synaptic transmission. In view of evidence that EAA transporters (EAAT) are heterogenous and contain consensus sites for phosphorylation, this investigation was undertaken to contrast the effects of transporter phosphorylation in fractions derived from glia and neurons (synaptosomes) of the adult rat forebrain. Treatment with phorbol-12,13 dibutyrate (PDBu), an activator of protein kinase C (PKC), increased the maximal rate of GLU transport in glial plasmalemmal vesicles by greater than 50 percent (237+/-18 vs. 365+/-27 pmol/mg protein/90s, p < 0.05) but caused no change in synaptosomes. The effect by PDBu was concentration and time-dependent and was inhibited completely by the PKC inhibitor calphostin C. Inhibition of serine threonine phosphoprotein phosphatases with okadaic acid produced similar effects which were not additive with PDBu. Together, these results demonstrate that glial EAAT can be regulated by multiple phosphorylation processes. PMID- 10478942 TI - Effects of endogenous glutamate on extracellular concentrations of GABA, dopamine, and dopamine metabolites in the prefrontal cortex of the freely moving rat: involvement of NMDA and AMPA/KA receptors. AB - Using microdialysis, interactions between endogenous glutamate, dopamine, and GABA were investigated in the medial prefrontal cortex of the freely moving rat. Interactions between glutamate and other neurotransmitters in the prefrontal cortex had already been studied using pharmacological agonists or antagonists of glutamate receptors. This research investigated whether glutamate itself, through the increase of its endogenous extracellular concentration, is able to modulate the extracellular concentrations of GABA and dopamine in the prefrontal cortex. Intracortical infusions of the selective glutamate uptake inhibitor L-trans pyrrolidine-2,4-dicarboxylic acid (PDC) were used to increase the endogenous extracellular glutamate. PDC (0.5, 2, 8, 16 and 32 mM) produced a dose-related increase in dialysate glutamate in a range of 1-36 microM. At the dose of 16 mM, PDC increased dialysate glutamate from 1.25 to 28 microM. PDC also increased extracellular GABA and taurine, but not dopamine; and decreased extracellular concentrations of the dopamine metabolites DOPAC and HVA. NMDA and AMPA/KA receptor antagonists were used to investigate whether the increases of extracellular glutamate were responsible for the changes in the release of GABA, and dopamine metabolites. The NMDA antagonist had no effect on the increase of extracellular GABA, but blocked the decreases of extracellular DOPAC and HVA, produced by PDC. In contrast, the AMPA/KA antagonist blocked the increases of extracellular GABA without affecting the decreases of extracellular DOPAC and HVA produced by PDC. These results suggest that endogenous glutamate acts preferentially through NMDA receptors to decrease dopamine metabolism, and through AMPA/KA receptors to increase GABAergic activity in the medial prefrontal cortex of the awake rat. PMID- 10478943 TI - Effects of lead on adenylate cyclase activity in rat cerebral cortex. AB - Lead decreased in a dose dependent manner the basal AC activity in membranes of rat cerebral cortex (IC50 = 2.5 +/- 0.1 microM). In membranes preincubated under basal conditions, AC activity was stimulated by approximately two and fourfold by 10 microM Gpp(NH)p or forskolin, respectively. Under basal conditions, lead (3 microM) inhibited enzyme activity up to 50%, but was not able to inhibit the Gpp(NH)p- or the forskolin-stimulated AC activity. However, in membranes preincubated with Gpp(NH)p (10 microM), lead (3 microM) had no significant effect on enzyme activity, but it partly blocked the stimulation of AC activity elicited by forskolin (10 microM). In membranes preincubated with 10 microM lead, the addition of 10 microM Gpp(NH)p or forskolin in the incubation medium did not stimulate AC activity. However, when added together in the incubation medium Gpp(NH)p + forskolin produced an increase in enzyme activity. In membranes preincubated with 10 microM lead + 10 microM Gpp(NH)p, Gpp(NH)p (10 microM) or forskolin (10 microM) added alone or in combination to the incubation medium did not stimulate AC activity. Moreover, under these latter conditions lead had no further effect on enzyme activity. These results indicate that lead may interact with G-proteins and with the catalytic subunit of cerebral cortical AC to produce inhibition of the enzyme activity. PMID- 10478944 TI - Release of GPI-anchored Zn2+-glycerophosphocholine cholinephosphodiesterase as an amphiphilic form from bovine brain membranes by bee venom phospholipase A2. AB - Enzymatic release of Zn(2+)-glycerophosphocholine (GPC)cholinephosphodiesterase, as an amphiphilic form, from bovine brain membranes was examined. Of various membrane hydrolases, bee PLA2 was the most effective in the release of the GPC cholinephosphodiesterase (amphiphilic form, 63-70%) from membrane. Compared to pancreatic PLA2, bee PLA2 was more efficient in the release of GPC cholinephosphodiesterase. In pH-dependent release of GP1-anchored phosphodiesterase, there was a similar pH-release profile between PLA2-mediated release and spontaneous one, implying the involvement of membrane disruption in the PLA2 action. The PLA2-mediated release showed a limited time-dependence (until 45 min) and a limited dose dependence (up to 3 units/ml), characteristic of a receptor-type binding. An ionic binding of PLA2 to membrane may be alluded from the interfering effect of anionic phospholipids on the PLA2 action. In support of an interaction between PLA2 and membrane glycoproteins, the PLA2 action was found to be blocked by lectins, wheat germ agglutinin or concanavalin A. In combination with detergent, the PLA2-mediated release was found to be enhanced synergistically by saponin, a cholesterol-complexing agent. Meanwhile, an additive interaction between PLA2 and lysolecithin suggests that PLA2 action is independent of lysolecithin. It is suggested that the binding of PLA2 to specific sites of membranes, probably rich in GPI-anchored glycoproteins, may be related to the facilitated release of GPI-anchored proteins as amphiphilic form. PMID- 10478945 TI - NADPH-diaphorase activity in normally developing and intracranially transplanted retinas. AB - The activity and distribution of nicotinamide dinucleotide phosphate diaphorase (NADPH-d), an enzyme that is widely distributed in the central nervous system and involved in the production of the free radical nitric oxide, were investigated histochemically in the normal developing and intracranially transplanted retinas. In the normal rat retina, NADPH-d activity was first detected in cells in the ganglion cells layer (GCL) and blood vessels on the first postnatal day (P0). A small but distinct population of NADPH-d positive cells were observed along the inner border of the inner nuclear layer at P7. NADPH-d positive sublaminae began to appear in the inner plexiform layer during the second postnatal week, and several strongly reactive sublaminae resembling those observed in the adult were observed by the fourth postnatal week. The overall spatio- temporal sequence of development of NADPH-d positive cells in the transplanted retina was similar to that of the normal retina, except a lack of reactive in the inner plexiform layer in more mature transplants as compared with normal retinas of corresponding ages. These results indicate that the time course of development and distribution of NADPH-d cells in early postnatal retina requires signals mainly of intraretinal origin and is independent of influence from the surroundings. While this finding is supportive to the notion that neurons that are rich in NADPH-d are resistant to injury or perturbation, the observation of a lack of well organized NADPH-d reactive sublaminae in the inner plexiform layer in older transplants suggests a possible alteration in the synaptic circuitry in the inner retina with increasing postgrafting survival time. PMID- 10478946 TI - Proportions of Ca2+ channel subtypes in chick or rat P2 fraction and NG108-15 cells using various Ca2+ blockers. AB - The proportions of calcium (Ca2+) channel subtypes in chick or rat P2 fraction and NG 108-15 cells were investigated using selective L-, N-, P- and P/Q- type Ca2+ channel blockers. KCl-stimulated 45Ca2+ uptake by chick P2 fraction was blocked by 40-50% using N-type Ca2+ channel blockers [omega-conotoxin GVIA, aminoglycoside antibiotics and dynorphin A(1-13)], but was not inhibited by P- or P/Q-type blockers (omega-agatoxin IVA or omega-conotoxin MVIIC). On the other hand, KCl-stimulated 45Ca2+ uptake by rat P2 fraction was blocked by 30 approximately 40% using P- or P/Q-type Ca2+ channel blockers, but was not inhibited by N-type Ca2+ channel blockers. The L-type Ca2+ channel blockers 1,4 dihydropyridines, diltiazem and verapamil, but not calciseptine (CaS), inhibited both KCl-stimulated 45Ca2+ uptake and veratridine-induced 22Na+ uptake by chick or rat P2 fraction with similar IC50 values. CaS did not have any effect on 45Ca2+ uptake by either chick or rat P2 fraction. In NG108-15 cells, CaS, omega agatoxin IVA and omega-conotoxin MVIIC, but not omega-conotoxin GVIA, inhibited KCl-stimulated 45Ca2+ uptake by 30-40%. Various combinations of these Ca2+ channel blockers had no significant additional effects in chick or rat P2 fraction or NG 108-15 cells. These findings suggest that KCl-stimulated 45Ca2+ uptake by chick or rat P2 fraction and NG 108-15 cells is a convenient and useful model for screening whether or not natural or synthetic substances have selective effects as L-, N-, P-, or P/Q- type Ca2+ channel antagonists or agonists. PMID- 10478947 TI - Study of adenosine A2 receptors in membrane preparations from optic tectum of chicks. AB - Binding properties of the subtypes of adenosine A2 receptors in membrane preparations and the effects of adenosine receptor ligands on cAMP accumulation in slices from the optic tectum of neonatal chicks have been investigated. [3H]2 [4-(2-p-carboxyethyl)phenylamino]-5'-N-ethylcarboxaminoadenosin e (CGS 21680), a selective ligand for adenosine A2a receptors, did not bind to optic tectal membranes, as observed with rat striatal membranes. CGS 21680 also did not induce cyclic AMP accumulation in optic tectum slices. However, 5'-N ethylcarboxamidoadenosine (NECA), 2-chloro-adenosine or adenosine induced a 2.5- to 3-fold increase on cyclic AMP accumulation in this preparation. [3H]NECA binds to fresh non-washed-membranes obtained from optic tectum of chicks, displaying one population of binding sites, which can be displaced by NECA, 8 phenyltheophylline, 2-chloro-adenosine, but is not affected by CGS 21680. The estimated K(D) value was 400.90 +/- 80.50 nM and the Bmax was estimated to be 2.51 +/- 0.54 pmol/mg protein. Guanine nucleotides, which modulate G-proteins activity intracellularly, are also involved in the inhibition of glutamate responses by acting extracellularly. Moreover, we have previously reported that guanine nucleotides potentiate, while glutamate inhibits, adenosine-induced cyclic AMP accumulation in slices from optic tectum of chicks. However, the guanine nucleotides, GMP or GppNHp and the metabotropic glutamate receptors agonist, 1S,3R-ACPD did not alter the [3H]NECA binding observed in fresh non washed-membranes. Therefore, the adenosine A2 receptor found in the optic tectum must be the adenosine A2b receptor which is available only in fresh membrane preparations, and its not modulated by guanine nucleotides or glutamate analogs. PMID- 10478948 TI - Kinetics of brain cholinesterase inhibition following metrifonate administration. AB - In previous metrifonate (MTF) studies, there has been evidence for a preferential functional effect of the drug in cortical but not in striatal regions. In the present study we investigated the kinetics of brain cholinesterase (ChE) inhibition following an acute administration of MTF (100 mg/kg) in various brain regions of young and old Fischer 344 rats. The main objective was to test the hypothesis that the functional regional selectivity, observed in previous studies, was correlated with the extent of ChE inhibition. Using Karnovsky's method for histochemical staining, the highest staining intensity in control rats was found in the striatum and hippocampus, compared to a low basal activity in the frontal and frontoparietal cortices. In the striatum of drug treated old rats, enzyme inhibition was somewhat greater than that found in young rats. However, in the hippocampus, four to eight hours following MTF administration, the inhibition was greater in young compared to old rats. The differences in the sensitivity of various brain regions towards MTF induced ChE inhibition could not be correlated with the regional variation of MTF functional effects. PMID- 10478949 TI - Drosophila choline acetyltransferase temperature-sensitive mutants. AB - We used the reverse transcription-polymerase chain reaction (RT-PCR) to amplify choline acetyltransferase (ChAT) mRNA fragments from two temperature-sensitive alleles of Drosophila melanogaster, Cha(ts1) and Cha(ts2). Single base substitutions in the mutants (T1614A in Cha(ts1) and G1596A in Cha(ts2)) would result in amino acid changes for ChAT protein (Met403Lys in Ch(ts1) and Arg397His in Cha(ts2)). These base substitutions were confirmed in mRNA extracted from homozygous mutants using a Single Nucleotide Primer Extension assay (SNuPE) and are sufficient to produce thermolabile enzyme. Our results indicate that these temperature-sensitive mutants are point mutations in the structural gene for ChAT. Using a quantitative SNuPE assay we also show that similar levels of Cha(ts) and wild type transcripts are present in heterozygous flies (Cha(ts1)/+ and Cha(Ts2)/+) at both restrictive and permissive temperatures. This contrasts with RNase protection assays of ChAT mRNA in homozygous mutant animals where the levels of mutant mRNA decrease at restrictive temperature. PMID- 10478950 TI - Intracellular GABA-activated in-->out permeation of chloride across the Deiters' neuron membrane: modulation by phosphorylating activities. AB - The modulation of intracellular GABA activated 36Cl- in-->out permeation across single Deiters' neuron membranes has been studied in a microchamber system. Addition of Mg2+/ATP on the membrane cytoplasmic side reduces strongly the GABA effect as does ATP alone. However, the greatest inhibition of the GABA effect is given by the addition of Mg2+ to the intracellular side buffer: a complete block of the stimulation by GABA of 36Cl- in-->out permeation. This is interpreted as due to the presence in this case of a constant concentration of exogenous Mg2+ acting together with endogenous ATP in the small cytoplasmic layer on the membrane inner side. The addition of ADP to Mg2+/ATP increases the inhibitory effect of the latter. This is presumably due to an extra increase of ATP, locally under the membrane, due to phosphorylation of ADP by endogenous phosphocreatine. Overall, the data confirm that phosphorylating conditions impair the intracellular GABA action on 36Cl- in-->out permeation. PMID- 10478951 TI - Molecular structure of 5-methyl thiophene acryloyl ethyl thiolester: a vibrational spectroscopic and density functional theory study. AB - Enzyme-substrate intermediates involving the acyl group 5-methyl thiophene acryloyl (5-MTA) bound to the active site of an enzyme via a sulfur or selenium atom have been characterized by Raman spectroscopy (e.g., J. D. Doran and P. R. Carey, Biochemistry 1996, 35, 12495-12502, and M. J. O'Connor et al., J Amer Chem Soc 1996, 118, 239-240). Raman difference spectra reveal the Raman spectrum of the acyl group in the active site and, in turn, these can be used to probe acyl group conformation and active site forces and interactions. In order to improve the understanding of the relationship between conformational states and vibrational spectra of 5-MTA thiolesters, calculations based on a density functional theory analysis are undertaken for 5-methyl thiophene acryloyl ethyl ester. The calculations provide the precise geometries and energies of rotomers of 5-MTA ethyl thiolester involving rotational isomerism about the C--C single bonds flanking the ethylenic linkage and the S--C bond linking the ethyl group to the sulfur atom. The calculations also provide the vibrational spectrum for each conformer and these predictions are compared with the experimental Raman an IR data for the thiolester in carbon tetrachloride. Modes are identified that can act as conformational markers for isomerism about the C--C and S--C2H5 single bonds. These findings are used to identify the two conformational states giving rise to the Raman spectrum of the 5-MTA-S-enzyme formed by the viral cysteine protease HAV-3C. PMID- 10478952 TI - Infrared spectroscopy of human tissue. V. Infrared spectroscopic studies of myeloid leukemia (ML-1) cells at different phases of the cell cycle. AB - Infrared spectra of myeloid leukemia (ML-1) cells are reported for cells derived from an asynchronous, exponentially growing culture, as well as for cells that were fractionated according to their stage within the cell division cycle. The observed results suggest that the cells' DNA is detectable by infrared spectroscopy mainly when the cell is in the S phase, during the replication of DNA. In the G1 and G2 phases, the DNA is so tightly packed in the nucleus that it appears opaque to infrared radiation. Consequently, the nucleic acid spectral contributions in the G1 and G2 phases would be mostly that of cytoplasmic RNA. These results suggest that infrared spectral changes observed earlier between normal and abnormal cells may have been due to different distributions of cells within the stages of the cell division cycle. PMID- 10478953 TI - Pharmacologic application of FTIR spectroscopy: effect of ascorbic acid-induced free radicals on Deinococcus radiodurans. AB - Fourier transform infrared (FTIR) spectroscopy was used as a convenient and easy to-run method to monitor radical-induced damage on the radiation-resistant Deinococcus radiodurans strain. Increasing concentrations of ascorbic acid added to the culture medium during the stationary phase produced striking changes in the infrared spectra. These changes especially occurred in the 1700-900 cm(-1) region, which is spectroscopically assigned to the amide I and II components, nucleotide bases, phosphodiester backbone and sugar rings, and were correlated with the oxidant effect of ascorbic acid. Thus, FTIR analysis allows a rapid characterization of the changes induced by ascorbic acid in the cell environment, which can be correlated in part with the generation of free radicals. Beyond a critical ascorbic acid concentration of 40 mM, these free radicals can cause severe damage to the biomolecular components, as soon as the antioxidant defenses of the bacterium are overwhelmed. PMID- 10478954 TI - Spectroscopic and ultrastructural comparative study of cystine calculi in humans and dogs. AB - The careful analysis of cystine calculi may be important to detect the presence of other urinary alterations (such as hyperuricosuria, hypercalciuria, hyperoxaluria or bacterial infections) that coexist with cystinuria in many patients. For this reason, in the present study, 14 human and 17 canine cystine uroliths have been studied by infrared spectroscopy (IR), scanning electron microscopy (SEM), and energy dispersive X-ray analysis (EDX). According to the infrared analysis, most of the human and canine stones were composed of nearly pure cystine. However, in these calculi of apparently pure cystine, the study by SEM and EDX showed in many cases the presence of small amounts of calcium apatite. The infrared study of several samples heated at 750 degrees C confirmed the presence of phosphate, when it was difficult to detect this component in the spectra of the original samples owing to band overlapping. Other components detected in varying proportions in cystine calculi were magnesium ammonium phosphate hexahydrate (struvite), calcium hydrogen phosphate dihydrate (brushite), calcium oxalate (mono and/or dihydrate) and, in one case, a drug (oxolinic acid). PMID- 10478955 TI - Raman and solid state 13C-NMR investigation of the structure of the 1 : 1 amorphous piroxicam : beta-cyclodextrin inclusion compound. AB - The results of a Raman and solid state 13C-NMR spectroscopic investigation aimed at studying the conformation of piroxicam (P) and its interaction with beta cyclodextrin (betaCD) in 1 : 1 amorphous PbetaCD inclusion compound are reported. The 1700-1200 cm(-1) FT-Raman and the 13C CP/MAS NMR spectra of 1 : 1 PbetaCD inclusion compound are discussed and assigned in comparison with those of the three main modifications of piroxicam (alpha, beta, and monohydrate). The FT Raman and 13C-NMR results show that in 1 : 1 PbetaCD inclusion compound piroxicam mainly assumes the zwitterionic structure typical of monohydrate, even if the presence of a different structure, that is, beta form, is not excluded. Piroxicam monohydrate, differently from alpha and beta forms, is characterized by a zwitterionic structure with an internal proton transfer and an increased charge delocalization, as shown by our spectroscopic results. The charge delocalization characteristic of this zwitterionic structure gives rise to the interaction with betaCD via electrostatic and hydrogen bonds. The possibility of a host-guest interaction between piroxicam and betaCD is not excluded; the guest molecule can be accommodated in betaCD cavity by interaction via hydrophobic bonds. PMID- 10478956 TI - Circular dichroism spectroscopy analysis of conformational transitions of a 54 base pair DNA duplex composed of alternating CGCGCG and TATATA blocks. AB - CD spectroscopy was used to analyze conformational properties of a self complementary 54-mer DNA composed of alternating (CG)3 and (TA)3 hexamers. NaCl induced the Z-form in poly(GC), but the 54-mer remained the B-form under the same conditions. The B-Z transition was induced only after the addition of NiCl2. However, the Z-form was adopted by the whole molecule, not by the (CG)3 blocks alone. Two orders of magnitude higher concentrations of NiCl2 were required to induce the Z-form in poly(AT). The Z-form was also induced in poly(GC) by CsF that switched poly(AT) into the X-form, which seems to be a solution counterpart of D-DNA. Under these conditions the CD spectrum of the 54-mer was consistent with the (TA)3 blocks being in the X-form and the (CG)3 blocks in the B-form. At high concentrations of ethanol or trifluoroethanol, poly(AT) was an A-form, while poly(GC) adopted either Z-form, A-form or Z'-form. At the high trifluoroethanol concentrations the 54-mer cooperatively switched into a conformation whose CD spectrum was most consistent with the A-form in the (TA)3 blocks and the Z'-form in the (CG)3 blocks. This suggests that the base pairs are tilted in the Z'-form as in the A-form. The present article illustrates that CD spectroscopy can provide interesting pieces of information about conformational isomerizations and coexistence of multiple conformations in DNA molecules containing blocks of different simple sequence repeats. PMID- 10478957 TI - Expression of laminin-5 in ameloblastomas and human fetal teeth. AB - Extracellular matrix proteins have been shown to play important roles in the cell migration and differentiation in both normal and pathological conditions. In the present study, we used immunohistochemistry and in situ hybridization to determine the distribution of laminin-5 in ameloblastomas and developing human teeth. In ameloblastomas, the immunoreaction for the laminin-5 gamma2 chain was confined to the tumor cells of the peripheral area. The staining reaction was variable, being mostly weak and fragmented in the basement membrane structures surrounding the neoplastic islands. Some peripheral epithelial cells and some invading small ameloblastoma cell islands showed intense intracellular staining for the gamma2 chain. Tumor cells in the proliferating areas of ameloblastomas expressed gamma2 chain mRNA. The laminin-5 gamma2 chain was located beneath the dental lamina and in the outer, but not in the inner, enamel epithelium of the developing teeth. During the early hard tissue apposition stage, intense staining for the gamma2 chain was confined to ameloblasts, which also gave a strong signal for gamma2 chain mRNA. These results suggest that laminin-5 may contribute to the infiltrative and progressive growing potential of ameloblastomas. During human tooth development, however, laminin-5 may participate in the terminal differentiation of ameloblasts and in enamel matrix formation. PMID- 10478958 TI - Immunohistochemical analysis of bcl-2 family proteins in benign and malignant ameloblastomas. AB - To study the possible roles of bcl-2 family proteins in oncogenesis and cytodifferentiation of odontogenic epithelium, the expression of bcl-2, bcl-x and bax proteins was examined immunohistochemically in tooth germs and various types of ameloblastoma. In ameloblastomas, bcl-2 and bcl-x proteins were expressed intensely in peripheral columnar cells and weakly in central polyhedral cells, while bax protein reactivity was low. In variants of ameloblastoma, no expression of these proteins was found in the keratinizing areas or granular cell clusters. Basal cell ameloblastomas showed intense reactivity for bcl-2 and bcl-x proteins, but slight or no reactivity for bax protein. These results indicate that in ameloblastomas the bcl-2 family proteins might function primarily as anti apoptotic factors, reflecting proliferative activity. Malignant ameloblastomas showed decreased reactivity for bcl-2 protein and increased levels of bax protein as compared with benign ones. PMID- 10478959 TI - Burning mouth syndrome: prevalence and associated factors. AB - Burning mouth syndrome (BMS) is characterized by a burning sensation in the oral cavity although the oral mucosa is clinically normal. The syndrome mostly affects middle-aged women. Various local, systemic and psychological factors have been found to be associated with BMS, but its etiology is not fully understood. Oral complaints and salivary flow were surveyed in 669 men and 758 women randomly selected from 48,500 individuals between the ages 20 and 69 years. Fifty-three individuals (3.7%), 11 men (1.6%) and 42 women (5.5%), were classified as having BMS. In men, no BMS was found before the age group 40 to 49 years where the prevalence was 0.7%, which increased to 3.6% in the oldest age group. In women, no BMS was found in the youngest age group, but in the age group 30 to 39 years the prevalence was 0.6% and increased to 12.2% in the oldest age group. Subjective oral dryness, age, medication, taste disturbances, intake of L thyroxines, illness, stimulated salivary flow rate, depression and anxiety were factors associated with BMS. In individuals with BMS, the most prevalent site with burning sensations was the tongue (67.9%). The intensity of the burning sensation was estimated to be 4.6 on a visual analogue scale. There were no increased levels of depression, anxiety or stress among individuals with more pain compared to those with less pain. It was concluded that BMS should be seen as a marker of illness and/or distress, and the complex etiology of BMS demands specialist treatment. PMID- 10478960 TI - Elevated production of salivary nitric oxide in oral mucosal diseases. AB - Nitric oxide (NO) is known to play an important role in biological systems. In this study, we measured levels of NO in the saliva of 39 patients with oral mucosal diseases: 21 had oral lichen planus (OLP) and 18 had recurrent aphthous ulceration (RAU). NO was assayed using the Griess reagent, which measures nitrite (NO2), the byproduct of NO. NO2 was detected in all tested samples, and levels in the saliva of patients were significantly increased relative to those of healthy subjects. We also examined the effect of NO on fibroblasts, keratinocytes and NA cells (an epithelial cancer cell line) in vitro. S-nitroso-N-acetyl-DL penicillamine (SNAP) and 3-morpholinosydnonimine (SIN-1) were used as NO donating reagents. The results revealed that cell viability was significantly reduced by NO derived from SNAP and SIN-1 in a dose-dependent manner. Although the role of salivary NO in normal physiology is as yet unknown, these findings suggest that excessive salivary NO plays a potential role in modifying oral mucosal diseases as a physiopathological regulator. PMID- 10478961 TI - Antioxidant status of oral mucosal tissue and plasma levels in smokers and non smokers. AB - The antioxidant status of an individual is thought to be important in the development of potentially malignant oral lesions (PMOL) and oral squamous cell carcinoma (OSCC). To date, little detailed information on mucosal antioxidant status is available in a United Kingdom population and neither has the relationship between smoking and mucosal antioxidant status been established. Furthermore, it has been implied that serum levels of antioxidants and tissue levels in the oral mucosa should be equivalent, but that is unproven. To address these deficiencies in our knowledge we studied 60 individuals, all of whom had an oral mucosal biopsy and simultaneous venous blood sampling. Antioxidant levels were measured using high performance liquid chromatography. Smokers (n= 19) were found to have significantly lower levels of plasma beta-carotene (P<0.05) and significantly lower levels of tissue alpha-carotene (P<0.05) than non-smokers (n=41). Tissue alpha-carotene correlated with plasma levels, but this was not the case with alpha-tocopherol, retinol, lycopene or beta-carotene. This is the first data on oral mucosal antioxidant levels and provides baseline data from which to study patients with potentially malignant oral lesions and oral squamous cell carcinoma. PMID- 10478962 TI - No evidence for specific in vitro lymphocyte reactivity to HgCl2 in patients with dental amalgam-related contact lesions. AB - Blood lymphocytes from 20 patients with oral contact lesions to dental amalgam and 10 healthy individuals were analyzed for HgCl2-induced proliferation in vitro, using both a modified assay and a conventional assay. The release of interferon-gamma (IFN-gamma) was measured in cell supernatants. Six patients displayed positive reactions in patch tests to mercuric compounds. No significant differences were recorded in HgCl2-induced proliferation in cells from patients and controls, since only few in the whole material responded to submitogenic concentrations. IFN-gamma was detectable in cell supernatants from some patients but also from controls and is not predictive of mercury allergy. Neither the phenotypes of peripheral lymphocyte subsets, the frequency of circulating cells expressing the interleukin-2 (IL-2) receptor, spontaneous lymphocyte proliferation nor concentrations of serum interleukin-6 differed between patient and control samples. In contrast to what has been claimed before, we did not find any evidence for specific in vitro lymphocyte reactivity in patients with oral contact lesions. PMID- 10478963 TI - Altered growth response of oral mucosal keratinocytes in p53-deficient mice. AB - P53 has important regulatory functions in cell growth, differentiation and apoptosis. Here we analyzed the effects of p53 on the growth response of oral mucosal keratinocytes (OMKCs) using p53-deficient (p53-/-) mice. No morphological difference was found between p53-/- and wild-type (p53+/+) oral mucosa. In a long term culture, p53-/- OMKCs continued to proliferate past the point at which p53+/+ became senescent. The percentage of p53-/- OMKCs in the G0/G1 phase was lower than that of p53+/+ OMKCs. Proliferation of cultured OMKCs induced by epidermal growth factor (EGF) and interleukin-(IL)-1alpha was more strongly enhanced in p53-/- than in p53+/+ mice. Such an enhanced response was not due to increased mRNA expression of growth factor receptors. These data suggest that p53 acts as a modulator of G1 arrest in OMKCs and is also involved in the regulation of responses to EGF and IL-1alpha without affecting the expression of their receptors. PMID- 10478964 TI - Simple bone cyst of the mandibular condyle with severe osteoarthritis: report of a case. AB - A rare case of simple bone cyst in the mandibular condyle of a 33-year-old woman is reported. The condition was difficult to diagnose because the cyst was accompanied by severe osteoarthritis. T1-weighted magnetic resonance (MR) images revealed a cystic lesion with intermediate signal intensity within the condylar head and an irregular margin with intermediate signal intensity on the superior surface of the condyle. The patient was treated by high condylectomy, discectomy and reconstruction by sagittal split ramus osteotomy. Histopathological examination showed a simple bone cyst in the condylar head and erosion of the anterior articular surface due to degenerative changes. PMID- 10478965 TI - Local tumor recurrence or emergence of a new primary lesion? A molecular analysis. AB - The distinction between a new primary oral tumor and recurrence may bear significant prognostic implications. Currently, this differentiation relies mainly on tumor location: when both lesions are at or near the same site, the new one is regarded as a recurrence; when the two are at different sites, the second lesion is regarded as a new primary. Recent investigations using molecular analysis have demonstrated that some oral squamous cell carcinomas (SCC) arising from different sites show the same clonogenical changes. In this case report, we studied the clonality of three SCC (one primary, two apparent recurrences) from the right lateral tongue of a young, non-smoking woman by using microsatellite analysis for loss of heterozygosity. The results showed that while the first two tumors were clonogenically similar, the third tumor was clonogenically different and was consistent with the development of a new primary. This result indicates that location of tumors alone is not always reliable in determining whether a new tumor is a recurrence or a new primary lesion. PMID- 10478966 TI - Contrast-enhanced power Doppler sonography for the differentiation of cystic renal lesions: preliminary study. AB - The objective of this study was to investigate the potential usefulness of contrast-enhanced power Doppler ultrasonography in the differentiation of benign and malignant cystic renal lesions. Our study group was limited to patients who had complex cystic renal lesions of uncertain malignancy at screening ultrasonography. During the previous 6 months, 485 patients have been referred to ultrasonography for evaluation of renal cystic lesions, but only 13 patients participated in this study. Focusing on tumor vascularity in the intracystic septa or solid component, we analyzed power Doppler sonographic images before and after intravenous injection of contrast agent and compared them with contrast enhanced CT scans or MR images and pathologic results. The visualization of vascularity was best on contrast-enhanced power Doppler sonography (n = 7). The use of contrast agent with power Doppler sonography showed improved diagnostic accuracy (77%) that was superior to non-contrast-enhanced power Doppler sonography or contrast-enhanced CT. In conclusion, contrast-enhanced power Doppler sonography provides better visualization of tumor vascularity in complicated cystic renal lesions than other imaging modalities, leading to more exact differential diagnosis. We therefore expect that this imaging modality might be very useful in differential diagnosis of problematic cystic renal lesions, benign or malignant. PMID- 10478967 TI - Sonographic appearance of early complete molar pregnancies. AB - Since our anecdotal experience indicates that the classically described "snowstorm" appearance on ultrasonography of early molar pregnancies is often not present and that theca-lutein cysts are also rare, we examined the ultrasonographic appearance of early complete molar pregnancies. We reviewed the ultrasonographic reports and clinical data of 21 cases of histologically diagnosed complete molar pregnancies with a mean gestational age at sonography of 10.5 weeks (range, 4 to 18 weeks). The diagnosis of molar pregnancy was made on ultrasonography in 12 (57%) cases, was second in the differential diagnosis of one (4.8%) case, and was not considered in eight (38%) cases. No theca-lutein cysts were identified. Five of five (100%) molar pregnancies of 13 weeks or over were diagnosed prospectively, while only eight of 16 (50%) earlier pregnancies were correctly diagnosed prospectively. In a retrospective review of the available images of 16 patients, only nine of 16 (56%) images demonstrated the classic appearance, and no theca-lutein cysts were seen. We conclude that the classic appearance of complete moles on ultrasonography is seen in less than two thirds of cases and even less commonly in the first trimester. The prevalence of theca-lutein cysts is very low. PMID- 10478968 TI - Extended field-of-view two-dimensional ultrasonography of the breast: improvement in lesion documentation. AB - The purpose of this study was to evaluate the use of extended field-of-view two dimensional ultrasonographic imaging for improvement in overall breast lesion documentation. Sonographic images of 59 patients with breast lesions or silicone implants were evaluated by three radiologists retrospectively to compare traditional static linear array images alone with images obtained with the addition of an extended field of view to determine if documentation of lesions was improved. The addition of extended field-of-view imaging improved lesion conspicuity by 21% over traditional images. It provided overall improvement in lesion documentation by including a reference point (nipple) or by more completely imaging large masses in 79% and implants in 69%. The larger field of view of this technique is promising as an adjunct to traditional sonography for breast lesion documentation. PMID- 10478969 TI - Stenosis of the main artery supplying an organ: effect of end-organ vascular compliance on the poststenotic peak systolic velocity. AB - Prior studies have shown variable results using poststenotic peak systolic velocity to detect hemodynamically significant renal artery stenoses. We postulated that vascular compliance, which affects the arterial waveform and varies by a factor of at least 5 in vivo, affects the peak systolic velocity, perhaps explaining the aforementioned variable results using peak systolic velocity to detect stenoses. A hydraulic model was used to investigate the relationship between end-organ vascular compliance and the peak systolic velocity. The peak systolic velocity was found to be mildly dependent on vascular compliance, decreasing with decreasing compliance. These results help explain some of the reported variability using peak systolic velocity to detect hemodynamically significant renal artery stenoses, but the effect is not great enough to explain the variability completely. Other factors not investigated in this study must exist that also affect peak systolic velocity. PMID- 10478970 TI - A case of an intratesticular leyomyoma. PMID- 10478971 TI - Interlaboratory comparison of ultrasonic backscatter, attenuation, and speed measurements. AB - In a study involving 10 different sites, independent results of measurements of ultrasonic properties on equivalent tissue-mimicking samples are reported and compared. The properties measured were propagation speed, attenuation coefficients, and backscatter coefficients. Reasonably good agreement exists for attenuation coefficients, but less satisfactory results were found for propagation speeds. As anticipated, agreement was not impressive in the case of backscatter coefficients. Results for four sites agreed rather well in both absolute values and frequency dependence, and results from other sites were lower by as much as an order of magnitude. The study is valuable for laboratories doing quantitative studies. PMID- 10478972 TI - Prevention of a first episode of variceal bleeding: role of duplex Doppler sonographic measurement of the acute response to beta-blockers. AB - The aim of our study was to assess whether acute variations in portal vein Doppler sonographic parameters induced by administration of a single beta-blocker agent are predictive of the long-term effects of these drugs in the prevention of a first episode of variceal bleeding. In 30 patients with liver cirrhosis at high risk for variceal bleeding, duplex Doppler sonographic parameters (maximal portal flow velocity, portal blood flow, and congestion index) were measured before and 4 h after the administration of 40 mg of propranolol. Twenty-three of these patients started chronic therapy with propanolol and were evaluated periodically (seven patients were excluded because they did not continue the therapy). The percentage of patients free from bleeding was 86.9% at the first year and 77.8% at the second year. Among a series of clinical, laboratory, and instrument-based parameters, the only one related to first bleeding, selected by the Cox regression model, was the percentage decrease in maximal portal flow velocity observed after initial administration of propranolol (P < 0.01). The best cutoff value for the percentage decrease in portal flow velocity (portal flow velocity test) was 12%. The prevalence of bleeding had been 25% (3 of 12) in patients with positive portal flow velocity test results (12% decrease or more), versus 64% (7 of 11) in patients with negative portal flow velocity test results. The actuarial probability of remaining free from bleeding (Kaplan-Meier analysis) was different in these two groups (log rank P < 0.01). The portal flow velocity test represents a safe and feasible method to predict the efficacy of beta-blockers in the prevention of a first bleeding episode in patients with cirrhosis. In patients with negative results on the portal flow velocity test, an alternative therapeutic approach should be considered. PMID- 10478973 TI - Sonographic diagnosis of ureteral tumors. AB - We present our experience with transabdominal ultrasonographic diagnosis of ureteral tumors. During the years 1989 to 1998, 16 patients were diagnosed as having ureteral tumors. These patients were referred for sonographic examination for evaluation of hematuria (seven patients) or flank pain (four patients) or for follow-up screening in patients who were asymptomatic but at high risk for transitional cell carcinoma because of known past bladder tumor (five patients). Ten of these patients underwent intravenous urography examination, three patients had retrograde pyelography, and 11 patients underwent CT scanning. Ultrasonography revealed the ureteral tumors in all 16 patients, which appeared as hypoechoic intraluminal soft tissue. Three tumors were localized in the upper ureter, four in the middle ureter, and nine in the distal ureter. The degree of ureterohydronephrosis was minimal (two cases), mild (five cases), moderate (eight cases), or severe (one case). Eleven tumors caused local widening of the ureteral diameter. On intravenous urography, four patients had a nonfunctioning kidney, three patients had unexplained ureterohydronephrosis, and three patients showed ureteral filling defects, of which only two had irregular contours. On retrograde pyelography, two patients had filling defects (one of which with smooth margins), and one had a truncated ureter. On CT the tumor was clearly demonstrated in only seven patients. We found that ultrasonography can be a useful diagnostic tool in the workup of ureteral tumors. PMID- 10478974 TI - Sonographic evaluation of the posterior cruciate ligament in amputated specimens and normal subjects. AB - The purpose of this study was to define the sonographic appearance and echogenicity of the normal posterior cruciate ligament. We examined the posterior cruciate ligament of five amputated specimens and five normal subjects using a 10 MHz linear array transducer. One K-wire was inserted into the substance of the posterior cruciate ligament of the amputated knee specimens to verify the location of the ligament on the sonogram. Various angles of insonation were used to examine the echogenicity of the posterior cruciate ligament. The results showed that the in situ posterior cruciate ligament appeared as a hypoechoic band relative to the surrounding tissue on sonograms, but it appeared hyperechoic when it was isolated and immersed in a water bath. The specific spatial orientation of the posterior cruciate ligament and anisotropy phenomenon contributed to the hypoechogenicity of the posterior cruciate ligament in situ on sonogram. PMID- 10478975 TI - Spinal sonography and magnetic resonance imaging in patients with repaired myelomeningocele: comparison of modalities. AB - The goals of this study were to evaluate the feasibility of using ultrasonography of the spine in the follow-up evaluation of patients with repaired myelomeningocele at birth and to compare sonography with the accepted modality of magnetic resonance imaging. Over a period of 4 years we performed 165 sonographic studies in 101 patients; 107 sonographic studies had MR imaging results for comparison. We collected our data prospectively. The quality of the sonograms was good in 110 of 129 studies, acceptable in 17 of 129, and poor in two of 129. The sonographic examinations failed in 33 of 165 studies (20%). Concordant information was obtained between ultrasonography and magnetic resonance imaging in the following percentage of studies: level of the distal end of the cord in 82%, position of the cord in the canal in 59%, presence of hydromyelia in 63%, cord duplication in 96%, adhesions in 16%, intradural mass in 37%, cord measurements in 85%, and dural sac measurements in 83%. At the lumbosacral level, we saw no cord pulsation in 57% of the studies in patients with cord adhesions and in 20% of those without adhesions. At the lower thoracic level, we saw no pulsation in 35% of the studies in patients with cord adhesions and in 7% of those without adhesions. Postoperative studies of cord release surgery in eight patients showed varied findings. We conclude that in those patients who have a spinal defect or interlaminar space allowing proper visualization of the lumbosacral spinal canal, ultrasound can provide fairly similar information to that obtained with magnetic resonance imaging of that area with no need for sedation and at a reduced cost. Ultrasonography seems more sensitive than magnetic resonance imaging in the detection of cord adhesions, which is particularly relevant in the diagnosis of tethering. PMID- 10478976 TI - Sonographic demonstration of a congenital laryngeal cyst. PMID- 10478977 TI - Healthy people report: progress toward year 2000 objectives, but some growing gaps. PMID- 10478978 TI - Task force 'grades' strategies for boosting vaccination rates. PMID- 10478979 TI - Guidelines for angina management released. PMID- 10478980 TI - Medications a problem in assisted living facilities. PMID- 10478981 TI - CDC issues recommendations on use of Lyme disease vaccine. PMID- 10478982 TI - NABP addresses pain management, confidentiality, Y2K preparedness. National Association of Boards of Pharmacy. PMID- 10478983 TI - FTC zeroes in on Internet health fraud. Federal Trade Commission. PMID- 10478984 TI - Warnings needed about PVC, group tells FDA. Plasticizer not harmful, says Koop team. PMID- 10478985 TI - Improving the prior-authorization process to the satisfaction of customers. PMID- 10478986 TI - Medicare drug benefit. PMID- 10478987 TI - Review of the Fifth American College of Chest Physicians Consensus Conference on Antithrombotic Therapy: outpatient management for adults. AB - The recommendations of the Fifth American College of Chest Physicians (ACCP) Consensus Conference on Antithrombotic Therapy are reviewed, with a focus on outpatient anticoagulation management in adults. Numerous therapeutic recommendations have changed since the Fourth ACCP Consensus Conference on Antithrombotic Therapy. The system of grading recommendations has been modified to emphasize clinically important differences and to take into account the benefit-risk ratio of each treatment option. The International Normalized Ratio (INR) goal is now expressed as a specific target value within a range rather than simply an INR range. The recommendations of the fifth conference cover initiation of warfarin therapy, hemorrhagic complications, management of excessive anticoagulation, interruption of warfarin therapy for patients requiring surgery, nonvalvular atrial fibrillation, cardioversion in patients with atrial fibrillation, valvular heart disease, mechanical and biological prosthetic heart valves, coronary artery disease, saphenous vein and internal mammary artery bypass grafts, peripheral arterial occlusive disease, prevention of venous thromboembolism, treatment of venous thromboembolism, stroke prevention in patients with cerebrovascular disease, and pregnancy. Since the fourth consensus conference, new anticoagulation therapies and indications have emerged; the recommendations of the Fifth ACCP Consensus Conference on Antithrombotic Therapy have provided practitioners with a resource of immense value. PMID- 10478988 TI - Direct health care costs for treatment of diabetes mellitus and hypertension in an IPA-group-model HMO. AB - The differences in direct health care costs and use between HMO enrollees with both diabetes mellitus and hypertension and enrollees with either disease alone were studied. Two years' worth of medical and pharmacy claims data from a hybrid (independent practice association and group)-model HMO were evaluated. Diagnoses were determined from medical claims data and cross-referenced with prescription information from pharmacy claims data. Aggregate costs associated with each disease, including pharmacy costs, costs of physician office visits, and laboratory costs, were compiled. Comparisons were made of all costs (any cost incurred by the health plan for the member, regardless of disease) and disease specific costs. The frequency of comorbid conditions was identified. A total of 6195 patients (670 with diabetes and hypertension, 1756 with diabetes alone, and 3769 with hypertension alone) were assessed. Patients with both diseases incurred much higher costs per year than patients with diabetes or hypertension alone (mean costs, $13,446, $8,493, and $8,424, respectively). Hospitalization costs contributed the greatest amount to total costs, while emergency room costs contributed the least. Disease-specific costs for diabetes and hypertension represented less than one quarter of total health care costs per patient. Average disease-specific costs were highest for patients with both diseases ($2,955), followed by costs for patients with hypertension alone ($1,803) and patients with diabetes alone ($689). The percentage spent on prescriptions was much higher for disease-specific costs than for total costs. The three most common comorbid conditions were dyslipidemia, coronary artery disease, and chronic obstructive pulmonary disease, with the frequency of cerebrovascular disease and myocardial infarction more than double in patients with diabetes and hypertension compared with patients with either disease alone. The cost of care for a patient with both diabetes and hypertension, although not double that for a patient with diabetes or hypertension alone, was higher than the cost of treating either disease. PMID- 10478989 TI - Azithromycin versus erythromycin for community-acquired pneumonia: a cost minimization analysis. AB - The costs of i.v. erythromycin versus azithromycin (in terms of medication use and treatment of adverse effects) when these drugs were used with other antimicrobials to treat community-acquired pneumonia (CAP) were compared. The medical records of patients receiving i.v. azithromycin or erythromycin as part of combination antimicrobial therapy for the treatment of CAP at a 473-bed level 1 trauma center in Kentucky were retrospectively reviewed. Data were collected for patients treated from December 1, 1997, through March 31, 1998. Patient data collected included occurrence of phlebitis or pain at the injection site, number of line changes due to phlebitis, and culture results. Cost data collected included drug acquisition cost, pharmacy cost of drug preparation, nursing time to administer the agent, cost of drug supplies, and cost of managing complications. Three time-and-motion studies were conducted to determine technician preparation time and pharmacist verification time. The medical records of 62 patients were identified and reviewed; 50 patients were enrolled in the study (25 in the azithromycin group and 25 in the erythromycin group). The average total days of therapy was 5.1 for the azithromycin group and 5.6 for the erythromycin group. The average total cost, including the cost of complications ($4.36 per patient in the erythromycin group), was $66.46 in the azithromycin group and $96.56 in the erythromycin group. The difference in costs between the two groups was not significant. There was no significant cost difference between azithromycin- and erythromycin-containing combination antimicrobial therapy in the treatment of CAP. PMID- 10478990 TI - Pharmacist recognition of potential drug interactions. AB - The ability of pharmacists to identify potential drug interactions was studied. Simulated medication profiles were created from a list of 16 drugs. Staff pharmacists and soon-to-graduate student pharmacists at a Veterans Affairs medical center each received a set of eight 2-drug profiles, four 4-drug profiles, two 8-drug profiles, and one 16-drug profile. Each set of profiles contained a number of pairs of drugs rated by the Drug Therapy Screening System as producing an interaction of moderate or major importance. The subjects were given one hour to screen the profile for the potentially interacting pairs. The subjects detected only 66% of the interactions in the 2-drug profiles, 34% of the interactions in the 4-drug profiles, 20% of the interactions in the 8-drug profiles, and 17% of the interactions in the 16-drug profile. None of the subjects detected all interactions in the 8- or 16-drug profiles. Both true positive and false-positive rates of identification decreased significantly as the number of drugs listed on the profile increased. This primarily reflected a reduced tendency to report the presence of drug interactions, but there was additional evidence that the accuracy of identification also declined. The number of years of pharmacy training was the only demographic characteristic highly correlated with accuracy. More years of pharmacy education seemed to improve the ability to detect drug interactions. However, none of the pharmacists or students was able to detect all potentially interacting pairs in a profile containing 8 or 16 drugs. Computerized drug interaction profiles should be used by pharmacists to ensure recognition of all potential drug interactions. PMID- 10478991 TI - Administration of crushed extended-release pentoxifylline tablets: bioavailability and adverse effects. AB - The pharmacokinetics of crushed and intact pentoxifylline tablets were compared, and the frequency of adverse effects was evaluated. Intact 400-mg extended release pentoxifylline tablets, crushed 400-mg tablets, intact 600-mg tablets, and crushed 600-mg tablets were given sequentially to 10 healthy male volunteers. Blood samples were collected at time 0, at 30-minute intervals for the first three hours, and at 4, 6, 8, 12, and 24 hours after the dose and analyzed by capillary gas chromatography for pentoxifylline and three major metabolites. The bioavailability of the crushed tablets relative to the intact tablets was 156% for the 400-mg strength and 137% for the 600-mg strength. The area under the plasma drug concentration-time curve from 0 to 24 hours (AUC0-24) for the 400-mg tablets (crushed and intact) differed significantly from that for the 600-mg tablets; there was no significant difference between intact 400-mg and intact 600 mg tablets or crushed 400-mg and crushed 600-mg tablets. The maximum plasma drug concentration (Cmax) was significantly greater and the time to maximum concentration (t(max)) significantly shorter for crushed tablets than intact tablets. The 400-mg crushed tablet caused mild nausea in three subjects. The 600 mg crushed tablet caused both moderate nausea and dizziness in seven subjects and diaphoresis, headache, and vomiting in one subject each. Cmax was higher and tmax shorter when pentoxifylline tablets were administered crushed rather than intact, and the increase in maximum plasma concentrations appeared to cause dose-related adverse effects; crushing the tablets did not decrease the relative bioavailability. PMID- 10478992 TI - Improving outcomes of community-dwelling older patients with diabetes through pharmacist counseling. PMID- 10478993 TI - Complication rates for a telephone-based anticoagulation service. PMID- 10478994 TI - Effect of an automated dispensing system on medication administration time. PMID- 10478995 TI - Distribution plan for immune globulin intravenous during a shortage. PMID- 10478996 TI - Home care pharmacist competency assessment program. PMID- 10478997 TI - Hydroxyurea in the treatment of human immunodeficiency virus infection. PMID- 10478998 TI - Finding drug link to reaction. PMID- 10478999 TI - Benefits of olanzapine as first-line schizophrenia therapy. PMID- 10479000 TI - Current literature. PMID- 10479001 TI - Coding for ureteroscopic procedures. PMID- 10479002 TI - Analgesia in piezoelectric SWL: comparative study of kidney and upper ureter treatments. AB - BACKGROUND: The development of newer-generation lithotripters has reduced the pain associated with SWL, but many patients still require some form of sedation. We prospectively compared the analgesic requirements for kidney and upper ureteral treatments. Predictive factors for pain during piezoelectric SWL were also studied. PATIENTS AND METHODS: A total of 102 consecutive patients without any previous experience of SWL were treated for renal (N = 70) or upper ureteral (N = 32) calculi using the EDAP LT02 lithotripter. The stones' largest diameter ranged from 4 to 30 mm (mean 9 mm). Patients were given an oral dose of 60 mg of dextropropoxyphene hydrochloride and 800 mg of paracetamol associated with 100 mg of ketoprofene per rectum 30 minutes before treatment. The SWL session was begun at low intensity and increased to the maximal range of energy as rapidly as could be tolerated by the patient. The amount of pain during treatment was recorded according to a visual analogue scale (VAS). Further analgesia using intravenous alfentanil was given as required by the severity of the pain. Visual analog pain scores, additional sedation requirements, and success rates after one session were analyzed. RESULTS: The VAS scores and intravenous sedation requirements were significantly lower for patients with upper ureteral stones than for those with renal calculi (P < 0.01). The stone-free rates after one session were, respectively, 90% and 73% (P < 0.05). On the other hand, SWL tolerance was significantly lower for women presenting with renal stones (P < 0.05). CONCLUSION: Piezoelectric SWL without intravenous sedation is suitable for the treatment of upper ureteral calculi. However, such an approach is less efficient in the management of kidney stones, especially for female patients. PMID- 10479003 TI - Prediction of stone composition from plain radiographs: a prospective study. AB - BACKGROUND AND OBJECTIVES: Stone composition, as reflected in radiographic appearance, is important to help choose between SWL and percutaneous/endoscopic procedures. Predicting a stone's composition accurately from a plain radiograph would be a useful tool in clinical decision-making. However, the ability of physicians to predict composition has not been adequately assessed. A prospective study was designed to quantify the accuracy of a panel of physicians who routinely deal with stones in classifying stone composition solely from radiographs. MATERIALS AND METHODS: A panel of six members was created to review 100 plain-film radiographs from patients with renal stones of known composition. The panel consisted of two urologists, two radiologists, and two nephrologists, all of whom have expertise in stone disease. If the composition guessed was at least 40% of the total stone composition, the response was deemed correct. RESULTS: Overall, there was an average 39% correct response score among the six panelists. When the stones were divided by size, 35% were <1 cm, and 65% were larger. The accuracy of chemical composition determination did not improve with greater stone size, nor was there a difference in accuracy for pure and mixed stones. The most frequently misclassified stone was calcium phosphate, with only 14% being correctly diagnosed. CONCLUSIONS: With a random sampling of plain radiographs, a panel of physicians specializing in stone disease correctly diagnosed the composition of renal calculi less than half of the time without being given clinical information. PMID- 10479004 TI - Effects of single-shot and twin-shot shockwaves on urinary enzyme concentrations. AB - BACKGROUND AND OBJECTIVE: Extracorporeal shockwave lithotripsy (SWL) remains the first-line treatment of urinary calculi. However, a number of studies have shown that adverse effects on the kidneys and the surrounding tissues may be encountered in short- and long-term follow-up. The aim of this study was to compare the effects of single-shot and twin-shot SWL techniques to identify the safest modality in terms of urinary enzyme excretion. PATIENTS AND METHODS: In this prospective, investigator-blinded, randomized study, urinary enzymes, beta2 microglobulin, microalbumin, Na, K, Ca, and creatinine concentrations were analyzed in 59 consecutive patients. Measurements were performed in urine specimens collected immediately before and after the SWL procedure and also on the 3rd and 7th days after treatment, which was performed on a Dornier MFL-5000 lithotripter utilizing the twin-shot technique (Group 1; N = 30) or the single shot technique (Group 2; N = 29) with 3000 shockwaves at 18 kV per treatment. RESULTS: Although there was no statistically significant difference in the results between the groups, urinary levels of microalbumin, alanine and aspartate aminotransferases, beta-2-microalbumin, gamma-glutamyltranspeptidase, Na, K, and Ca rose acutely after SWL, reaching maximum levels on the 3rd day, and returned to the baseline by the 7th day following the treatment in both groups. CONCLUSION: This study demonstrates that SWL performed by either a single-shot or twin-shot shockwave technique has a transient detrimental effect on renal function, as assessed by urine enzyme concentrations. It is recommended that the twin-shot shockwave technique be used in routine lithotripsy in consideration of the cost-effectiveness provided by the shorter treatment time. PMID- 10479005 TI - Cardiac dysrhythmias induced by extracorporeal shockwave lithotripsy. AB - PATIENTS AND METHODS: We evaluated in 269 consecutive patients the incidence and gravity of dysrhythmic complications during nonsynchronized extracorporeal shockwave lithotripsy (SWL) using an electromagnetic lithotripter. RESULTS: Dysrhythmia occurred during treatment in 22 patients (8.8%) with no previous cardiac dysrhythmia. Ventricular extrasystoles occurred in 14 patients, atrial extrasystoles in 7 patients, and sinus bradycardia in 1 patient. It was not necessary to terminate treatment because of the occurrence of dysrhythmia in any of the patients. For 13 of the 22 patients (59%), it was sufficient to interrupt the treatment momentarily to obtain resumption of the normal rhythm. For 8 patients (36%), treatment was continued after triggering the release of the shockwaves with the refractory phase of the heart cycle. For one case of bradycardia (42 beats/min), it was possible to continue with the treatment after intravenous administration of atropine 0.5 mg. Pretreatment dysrhythmias were revealed by the electrocardiographic examination in 16 of the patients studied (6.3%). CONCLUSIONS: Extracorporeal shockwave lithotripsy without ECG triggering has been found to be fast and efficient and not correlated with the occurrence of dysrhythmic episodes of any particular clinical significance. No significant correlation was found between the occurrence of dysrhythmia, the side treated, the number and strength of the shockwaves, or the administration of analgesics. It was found, however, that dysrhythmia occurred almost exclusively in treatments involving the kidneys. The ECG-triggering option was indispensable in some patients in order to complete the lithotripsy without complications. PMID- 10479006 TI - Symptomatic ureteropelvic junction (UPJ) obstruction. PMID- 10479007 TI - State-of-the-art magnetic resonance imaging of the kidneys and upper urinary tract. AB - State-of-the-art magnetic resonance imaging (MRI) is an excellent examination in several clinical contexts of interest to endourologists. First, it offers a one stop imaging examination of prospective renal donors, obviating the need for arteriography and conventional excretory urography. Second, it reliably depicts urinary tract obstruction and can usually distinguish acute from chronic obstruction, although it is not as sensitive as helical CT in detecting small, nonobstructing stones. Third, it is an excellent examination for characterizing renal masses, especially complex cystic masses, and for surgical planning. Because MRI does not use ionizing radiation and because gadolinium contrast agents are essentially non-nephrotoxic, MRI is especially useful in children, women of childbearing age, and patients with renal insufficiency or renal allografts. Future developments will likely include MR "fluoroscopy," which will provide real-time imaging guidance for interventional procedures in the urinary tract. PMID- 10479008 TI - Retroperitoneoscopic nephrectomy for benign diseases of the kidney: prospective nonrandomized comparison with open surgical nephrectomy. AB - PURPOSE: To describe, define, and evaluate the efficacy of retroperitoneoscopic nephrectomy (RPN) for benign diseases of the kidney and to compare it with open surgical nephrectomy (OSN) via a flank approach. PATIENTS AND METHODS: From August 1995 to November 1997, 29 men and 14 women (mean age 33 years) with severely damaged kidneys underwent RPN. Among these, 11 patients had undergone prior surgery, 3 had chronic renal failure, and 8 patients had a percutaneous nephrostomy. The RPN was performed via three or four ports, with the kidneys being removed intact from the retroperitoneal working space. During the same period, 43 patients underwent OSN through a flank approach (extrapleural and extraperitoneal) for nonfunctioning or poorly functioning kidneys. RESULTS: In the RPN group, two patients required conversion to OSN. The operative time and estimated blood loss ranged from 40 to 210 minutes (mean 114 minutes) and 50 to 450 mL (mean 127 mL), respectively. In the OSN group, the corresponding values were 60 to 100 minutes (mean 104 minutes) and 70 to 600 mL (mean 266 mL), respectively. The mean length of hospitalization after RPN was considerably shorter--2 to 7 days (mean 3.4 days)--than after conventional open surgery--4 to 16 days (mean 8.6 days). The incidences of minor and major complications were 21% and 5%, respectively, in the RPN group and 33% and 2% in the OSN group. The postoperative analgesic requirement was significantly less (P < 0.001) in RPN group. The interval to return to normal activity ranged from 7 to 30 days (mean 20.3 days) and 20 to 60 days (mean 32.9 days) in the RPN and OSN group, respectively, with superior performance status, cosmesis, and quality of life observed in the former group. CONCLUSION: Retroperitoneoscopic nephrectomy is as effective as open nephrectomy for benign kidney diseases with less postoperative pain, a shorter hospital stay, earlier recuperation, and excellent cosmesis. This procedure can also be performed in patients who have undergone abdominal operations previously, in those with chronic renal failure, and in those with a percutaneous nephrostomy. The operation has become our first line of approach for benign diseases of the kidney. PMID- 10479009 TI - Routine radiologic surveillance for obstruction is not required in asymptomatic patients after ureteroscopy. AB - OBJECTIVE: A retrospective medical record review was performed with the goal of determining the need for radiologic evaluation after ureteroscopy. PATIENTS AND METHODS: Of 183 patients undergoing ureteroscopic procedures at our institution between 1989 and 1993, 131 underwent postoperative radiologic studies capable of diagnosing ureteral obstruction. Of these patients, 110 (84%) were asymptomatic after the procedure, and radiologic procedures capable of displaying obstruction were performed at a median of 60 days (mean 73 days; standard deviation 189 days) after ureteroscopy. RESULTS: None of these asymptomatic patients displayed obstruction at the time of the routine follow-up radiologic procedure. Of those 21 patients (16%) who experienced flank pain subsequent to ureteroscopy, 13 were found to have ureteral obstruction secondary to ureteral calculus. One patient (1/131 or 0.8%) was found to have a ureteral stricture, which occurred after a full-thickness ureteral injury. All cases of postoperative obstruction were heralded by a concomitant display of flank pain. CONCLUSION: Routine postoperative radiologic studies are not necessary in surveillance for obstruction in the asymptomatic postureteroscopy patient, as obstruction should become evident by virtue of flank pain. The exception to this practice may be in patients experiencing a ureteral perforation intraoperatively, who may be at greater risk of stricture. PMID- 10479010 TI - Recurrent suture urolithiasis 29 years after open pyelolithotomy. AB - A case of a foreign body-induced renal stone in which the patient was treated successfully with a ureterorenoscopic approach is presented. PMID- 10479011 TI - Is antegrade ureteroscopy better than retrograde ureteroscopy for impacted large upper ureteral calculi? AB - OBJECTIVE: This study was planned to compare the risks and advantages of antegrade and retrograde ureteroscopy for impacted large upper ureteral calculi. PATIENTS AND METHODS: From September 1996 to February 1998, ureteroscopy was offered to 43 patients. Of these, retrograde ureteroscopy was done in 20 patients, while antegrade ureteroscopy was performed in 23 patients. All these patients were followed up to evaluate the immediate and long-term success of the procedure. The incidence and nature of complications were also noted. RESULTS: During retrograde ureteroscopy, complete stone clearance was achieved in 11 patients (55%), while pushback of the whole or fragmented calculus was seen in the rest. These patients with residual calculi were later treated by extracorporeal shockwave lithotripsy (SWL). The stone-free rate at the end of 3 months was 85%. Three patients developed minor ureteroscopy-related complications. Complete stone clearance was achieved in all patients with antegrade ureteroscopy. No intraoperative or postoperative complications were encountered. CONCLUSION: In this series, antegrade ureteroscopy was found to be a safe and effective option for impacted upper ureteral calculi and assured better results than retrograde ureteroscopy. PMID- 10479012 TI - Ureteropelvic junction obstruction presenting as early satiety and weight loss. AB - We present a case of ureteropelvic junction (UPJ) obstruction which had the unusual presentation of early satiety and weight loss secondary to gastric compression by a distended renal pelvis. The patient was treated successfully with percutaneous antegrade endopyelotomy. PMID- 10479013 TI - Box stitch modification of Hasson technique for pediatric laparoscopy. AB - Laparoscopy has been recognized as the modality of choice for the treatment of many diseases in urology, gynecology, and general surgery. Although laparoscopy is minimally invasive, the most frequent complications are associated with entry through the abdominal wall. The Hasson (open) technique was developed to minimize the complications by entering the abdomen under direct vision. We have modified the Hasson technique with a stitch that is easy to perform, creates a tight fascial seal, allows easy exchange of trocars, and produces an effect that is cosmetically appealing. PMID- 10479014 TI - Three-dimensional ultrasound and magnetic resonance imaging of pelvic anatomy: potential for complications from minimally invasive procedures. AB - OBJECTIVE: Several new minimally invasive therapies have recently been popularized for both malignant and benign prostate disorders, including interstitial implantation of radioactive seeds and high-radiofrequency wires, cryoablation, transurethral thermotherapy, and laser prostatectomy. Complications can be incurred during the various procedures, often as a result of injury to adjacent anatomic structures. Some of the complications are inadvertent, whereas others are inherent in the particular treatment process. We hope to increase awareness and understanding of some of the potential complications. METHODS AND MATERIALS: Magnetic resonance (MR) and three-dimensional transrectal ultrasonography (TRUS) imaging were utilized to illustrate the relevant pelvic anatomy in, respectively, a healthy volunteer and four patients undergoing evaluation for prostate symptoms. In addition, data from the Visible Human dataset (the Visible Human Project is part of the National Library of Medicine 1986 Long-Range Plan) were used. RESULTS: The potential complications relating to urinary sphincter and anal sphincter control, sexual function, pelvic musculature, and pelvic nerve physiology could be explained on the basis of the MR and TRUS findings using cryoablation for illustrative purposes. CONCLUSION: A clear understanding of the relevant anatomy and physiology is essential for the physician to provide patient counseling preoperatively regarding anticipated sequelae and to avoid preventable intraoperative complications related to minimally invasive therapeutic procedures for the prostate. PMID- 10479015 TI - Vigilance transitions in reaction time test: a method of describing the state of alertness more objectively. AB - Electrophysiological recordings are considered a reliable method of assessing a person's alertness. The aim of this study was to show, firstly, that changes in alertness during a Reaction Time Test (RTT) can be determined with certain adaptive scoring stages but not with R&K scoring and secondly, that the different adaptive stages can explain findings in reaction time. In 17 male patients (50.8+/-9.7 years, Body-Mass Index (BMI) 31.9+/-5.1 kg/m2) diagnosed with Obstructive Sleep Apnea Syndrome (OSAS) (Respiratory Disturbance Index (RDI) 53.3+/-24.1 /h sleep) a 90 min daytime vigilance test was performed twice, after the diagnostic polysomnographic investigation and after two nights spent with nCPAP. After a computerised adaptive segmentation analysis, a visual rule-defined classification system categorised alertness into one of 12 adaptive scoring stages. 6 of the 12 stages are described by the alertness conditions comparable to WAKE and NREM1.4 stages are nearly classified as NREM2-4, Rapid Eye Movement (REM) and Movement Time (MT), and one stage reflects the increase of alertness from drowsiness. The typical stage of an alert subject increased significantly from a median of 65.9% before therapy to 80.8% in the second investigation. The percentages of clearly drowsy stages decreased significantly. In contrast, there were no significant changes in the percentages of sleep stages according to R&K criteria for both investigations. According to R&K criteria 178 of 398 failed reactions (Reaction time >10 s) occurred in stage WAKE. According to adaptive scoring, only 12 failed reactions appeared in the alert stage. During the other failed reactions the electrophysiological recordings showed decreases in alertness. Neither the visual assessment nor the descriptive statistical results of R&K scoring were helpful to interpret the patient's alertness condition. In contrast, the patients' increases in alertness with nCPAP could be described by the adaptive scoring stages. This method could be a very useful procedure, when an expert opinion is necessary. It also has an actual context to the discussion of the effectiveness of CPAP in the treatment of OSAS. PMID- 10479016 TI - Constantly discontinuous EEG patterns in full-term neonates with hypoxic ischaemic encephalopathy. AB - OBJECTIVES: Selected EEG features were evaluated in 21 constantly discontinuous tracings recorded on the same number of full-term neonates with hypoxic-ischaemic encephalopathy. METHODS: The tracings were examined without using interval amplitude as the basis for distinguishing between burst-suppression and nonburst suppression patterns. RESULTS: The results were related to outcomes and other clinical parameters (severity of hypoxic-ischaemic encephalopathy, pO2 levels and drug intake). CONCLUSIONS: Features defining the grade of EEG discontinuity (i.e. maximum interval duration, minimum burst duration and interval amplitude) significantly related to outcome and, in most cases, to the grade of hypoxic ischaemic encephalopathy. Other features (amplitude of slow waves within the burst and incidence of abnormal EEG transients) related to PO2 levels. The consumption of anticonvulsant drugs increased EEG discontinuity, but this effect did not seem dose-related. Finally, the persistence of a constantly discontinuous EEG pattern after the first week of life is a sign of unfavourable prognosis. In full-term neonates with hypoxic-ischaemic encephalopathy quantitative analysis of all constantly discontinuous EEGs seems more useful than only describing burst suppression patterns on the basis of interval amplitude. PMID- 10479017 TI - The periodicity of lateralized epileptiform discharges. AB - OBJECTIVES: Although the term periodic lateralized epileptiform discharges (PLEDs) was introduced in 1964, there has been little discussion about the definition of periodicity, whether the degree of periodicity has clinical significance, and whether electroencephalographers can accurately assess periodicity. The objectives of this study were to determine if a mathematical descriptor of periodicity could analyse PLEDs and whether such has clinical relevance, to correlate the degree of periodicity with underlying etiology and to assess the accuracy of sight recognition of periodically occurring EEG events. METHODS: A 6 year retrospective analysis of EEGs on patients with PLEDs was undertaken. The coefficient of variation of inter-complex intervals was calculated for all records. The relative periodicity of each record was rated by electroencephalographers. Corresponding hospital records were reviewed with regard to clinical and pathological information. RESULTS: In cases of acute viral encephalitis, discharges were more periodic than in other etiologies (P = 0.013). Age, clinical state, and time of last seizure did not influence periodicity but 40 (70%) of 57 recordings with PLEDs were done within 2 days of a seizure. A history of seizures was present in 42 (89%) of 47 patients. Electroencephalographers' assessment of periodicity correlated well with measured periodicity (r = -0.49, P = 0.0001). However, EEGers were more likely to assess records as periodic if: discharges occurred frequently (r = -0.48, P = 0.0001), or the complexes were visually assessed to have a uniform morphology (r = 0.31, P = 0.02), even though neither of these factors influenced the periodicity of discharges. CONCLUSIONS: PLEDs were objectively measured to be more periodic when they were associated with acute viral encephalitis than with other etiologies. Nonetheless, the considerable overlap of values among the several presenting disorders diminished specificity. Electroencephalographers were able to recognize periodically occurring discharges. However, they were strongly influenced by discharge frequency and their perception of the uniformity of discharge morphology. PMID- 10479018 TI - Cortical reflex myoclonus studied with cortical electrodes. AB - OBJECTIVE: To study the mechanism of cortical reflex myoclonus. METHODS: A patient with stimulus sensitive myoclonus of the left foot had an array of subdural electrodes placed over the right sensorimotor cortex. RESULTS: Stimulation through one of the electrodes (contact 13) facilitated leg muscles with the shortest latency and was presumed to lie over the motor cortex. Tibial nerve stimulation evoked a potential with the shortest latency 1 cm further posteriorly (contacts 11-12). These contacts were presumed to lie over the sensory cortex. The potential at 11-12 was followed by a much larger potential that reversed polarity at contact 13. Back averaging from spontaneous myoclonic jerks showed a cortical premovement potential which reversed polarity at contact 13. The threshold for the motor evoked potential in leg muscles evoked by transcranial magnetic stimulation was lower on the affected side. Electrical stimulation through contact 13 produced cortical potentials that could be recorded at adjacent contacts. The combination of a positive potential followed by a negative potential recurred at approximately 35-40 ms intervals, each positive potential generating a myoclonic jerk. Additional waves resembling I waves accompanied only the first positive potential. Surgical removal of the cortex under electrode 13 abolished the myoclonus. CONCLUSIONS: The myoclonic jerks arose in the motor cortex. We postulate that there is increased excitability or synchronization of the cortical neurons at that site. The spontaneous, peripherally induced and recurrent cortical potentials and myoclonic jerks can occur without participation of the circuitry of the presumed I waves. PMID- 10479019 TI - Repeated nocturnal sleep latencies in narcoleptic, sleepy and alert subjects. AB - OBJECTIVES: The purpose of this study was to assess nocturnal sleep latencies among narcoleptics. METHODS: Thirteen narcoleptics and matched sleepy and alert controls participated in this study. Subjects were awakened three times on each of two experimental nights. The latencies to sleep and rapid eye movement sleep were evaluated at the beginning of the night and following each experimental awakening. RESULTS: The alert group (AG) had a significantly longer mean nocturnal sleep latency than the narcoleptic (NG) and sleepy groups (SG). The sleep latencies at 23:00 and 01:10 h were significantly longer than the latencies at 03:10 and 05:10 h. The interaction between group and time of night demonstrated longer latencies at 23:00 and 03:10 h for the AG when compared to the SG and the NG. At 01:10 and 05:10 h all groups had comparable latencies. The number of subjects in the NG who had multiple sleep onset REM periods (SOREMPs) was significantly higher than in either the AG or the SG. CONCLUSIONS: Narcoleptics were found to have a heightened propensity to fall asleep and increased number of SOREMPs during nocturnal sleep opportunities. These characteristics are consistent with the daytime polysomnographic findings known in this patient population. PMID- 10479020 TI - Origin and propagation of spontaneous electrographic sharp waves in the in vitro turtle brain: a model of neuronal synchronization. AB - OBJECTIVES: Neuronal synchronization is a basic feature in the generation of epileptiform discharges. Spontaneous large sharp waves (LSWs) can be recorded in the turtle brain in vitro, indicating the synchronous activation of large neuronal populations. The aim of this study was to analyze the spatial and temporal distribution of LSWs within the brain; the participation of glutamate in LSWs generation was also investigated. METHODS: Extracellular field potentials were recorded in vivo (n = 4) and in vitro (n = 36). LSWs were recorded from cerebral cortex, optic tectum, and thalamus. RESULTS: LSWs were recorded from cerebral cortex, optic tectum and thalamus. No LSWs were observed in cerebellum and brain stem. In some experiments, LSWs could be recorded only from medial cortex. Latency studies demonstrated that, within each hemisphere, medial cortex led the generation of LSWs; in addition, isolated medial cortex could sustain LSWs. Intracortical laminar field potentials in medial cortex indicated that LSWs generate mainly in the molecular layer, probably at pyramidal cell dendrites. Pharmacological experiments demonstrated that NMDA and non-NMDA glutamate receptors are involved in LWSs generation. CONCLUSIONS: These results suggest that turtle medial cortex is the pacemaker area for LSWs generation and it can be a useful model to study cellular and circuital mechanisms of neuronal synchronization. PMID- 10479021 TI - Development of a silicon retinal implant: cortical evoked potentials following focal stimulation of the rabbit retina with light and electricity. AB - There are currently many experimental efforts to develop a treatment for retinitis pigmentosa (RP) and age-related macular degeneration (AMD). The efforts reported here are part of the Retinal Implant Project, a collaborative effort of the Massachusetts Eye and Ear Infirmary and the Massachusetts Institute of Technology, to develop an independent functioning epiretinal prosthesis to restore vision in patients with disease of the outer retina such as RP and AMD. The epiretinal prosthesis will electrically stimulate the inner retina, bypass the damaged photoreceptors, and hopefully result in meaningful vision. One way to monitor the epiretinal implant is to record focal (stimulation of a small area of retina) evoked cortical potentials (EEP) upon electrical current stimulation. EEP recordings have 3 uses: (1) verify that focal retinal electrical stimulation sends a signal to the cortex that is similar to that elicited by light; (2) develop an animal model to chronically monitor the epiretinal implant; and (3) investigate optimal stimulus parameters. Five dutch pelted rabbits were placed under anesthesia and a 250 microm concentric bipolar stimulating electrode was introduced on the vitreal surface of the retina under visual guidance to record the EEP. Stimuli were charge-balanced pulses and recording electrodes were extradural. Focal VEPs were also recorded and compared to the focal EEP to ascertain cortical origin of the EEP, determine similarities of the EEP to the VEP and determine focality of electrical stimulus. EEP recordings were elicited in 3 out of the 5 rabbits. Current amplitudes which produced detectable responses ranged from 50-1000 microA. In our best series, amplitudes increased by 47-206 microV in increasing current from 50-600 microA. In comparing latencies of the focal EEP and focal VEP, the EEP latencies were 60-70 ms faster, favoring a cortical signal origin. In comparing amplitudes, VEP and EEP responses behaved similarly with changes in stimulus strength and stimulating frequency. In conclusion, EEPs were obtained with parameters of electrode size and current threshold level that would be used by the epiretinal implant, enabling use of the EEP response to monitor the epiretinal implant. PMID- 10479022 TI - The fundamental and second harmonic of the photopic flicker electroretinogram: temporal frequency-dependent abnormalities in retinitis pigmentosa. AB - OBJECTIVES: The flicker electroretinogram (FERG) consists mainly of a linear (fundamental, 1F) and a non linear (second harmonic, 2F) component. Previous results indicate that 2F originates more proximally in the retina than 1F, and that retinitis pigmentosa (RP) may affect 2F to a greater extent than 1F. The aim of this study was to evaluate FERG 1F and 2F abnormalities in RP as a function of the stimulus temporal frequency (TF). METHODS: Twelve patients with typical RP and 10 age-matched controls were examined. FERGs were recorded in response to uniform fields (18 degrees) presented in the macula on a light-adapting background. Stimuli were flickered sinusoidally at different, closely spaced TFs between 3.7 and 52 Hz. Amplitudes and phases of the Fourier analysed 1F and 2F components were measured. Components' apparent latencies were estimated from the rate at which phase lagged with TF. RESULTS: When compared to controls, mean 1F amplitudes of patients were reduced at both low (3.7-12.6 Hz) and high (14-52 Hz) TFs, with greatest losses (0.5 log units) around the peaks (3.7 and 41 Hz) of the normal TF function. Mean 2F amplitudes were reduced mainly at low TFs, with greatest losses (0.5 log units) at 5-8 Hz. On average, the shape of the 2F, but not 1F amplitude versus TF function, differed between patients and controls, showing a selective attenuation at low TFs. Mean 1F apparent latencies were delayed at both low and high TFs, with greater delays at low (85 ms) than at high (33 ms) TFs. Mean 2F apparent latencies were delayed only at low TFs (58 ms). CONCLUSIONS: In RP, 1F and 2F phase delays as well as 2F amplitude losses are dependent on TF, suggesting that FERG generators' subpopulations in both distal and proximal retina are differentially affected. Analysis of the FERG TF response is potentially useful to characterize cone system dysfunction in different genetic subtypes of RP. PMID- 10479023 TI - Individuals with Down's syndrome demonstrate abnormal olfactory event-related potentials. AB - OBJECTIVE: Recent research has demonstrated that individuals with Down's syndrome (DS) develop plaques and tangles in the brain similar to people with Alzheimer's disease. As a result, they show increased dementia and decreased olfactory functioning compared to healthy individuals. The olfactory event-related potential (OERP) has been used as an objective quantitative measure of olfactory functioning in normal and clinical populations. The present study investigated the utility of the latency and amplitude of the OERP components in examining olfactory dysfunction in DS individuals. METHODS: OERPs were recorded monopolarly at the Fz, Cz and Pz electrode sites, using amyl acetate at a 60 s inter-stimulus interval, from individuals with DS (mean age 26.0 years) and age-matched normal controls. Participants were screened for nasal health and odor thresholds were assessed. Dementia was assessed using the dementia rating scale (DRS). RESULTS: Results indicate that DS subjects have significantly longer latencies in the sensory (N1, P2, and N2) and cognitive (P3) components of the OERP than normal controls. Odor threshold was significantly associated with sensory OERP components. In addition, DS subjects with a higher level of dementia showed significantly longer P3 latencies than those with lower dementia levels. CONCLUSIONS: The study suggests that the OERP may be a useful measure of olfactory dysfunction in DS which may precede developing dementia in this population. PMID- 10479024 TI - Event-related brain potentials as indicators of smoking cue-reactivity. AB - OBJECTIVE: Reactivity to smoking cues, shown previously by autonomic and self report variables, was investigated in smokers and nonsmokers using event-related brain potentials (ERPs). METHODS: Average ERPs to 20 color pictures of people smoking and 20 neutral pictures depicting nonsmoking themes, randomly mixed with 4 repetitions/stimulus, were measured from 20 light-moderate smokers and 18 nonsmokers, following a stressor. Smoker status and stimulus type effects on ERPs and principal components factor scores (FS) were tested by repeated-measures ANOVAs. RESULTS: Smokers' N268 showed significant medial and midline smoking cue reactivity (ERP to smoking-related minus neutral stimuli); while a P300-like, P412, showed significant smoking cue-reactivity over medial and left hemisphere scalp. FS analyses confirmed most of the foregoing. P412 smoking cue-reactivity was correlated with unpleasantness-pleasantness cue-reactivity but not with urge to-smoke cue-reactivity. Nonsmokers' N268 stimulus differences were not significant, but significant P412 stimulus effects (unconfirmed by FS analyses) were found in central-to-posterior and in left parietal-temporal areas. CONCLUSIONS: Smokers' N268 is identified with a process detecting stimuli incongruent with tobacco-addicted states; and P412 smoking cue-reactivity is discussed in terms of an automatic, perceptual-categorization system, consistent with Tiffany's drug-use and Johnson's P300 models. Implications of ERP smoking cue-reactivity for study of tobacco and other addictions are discussed. PMID- 10479025 TI - Neural source estimation from a time-frequency component of somatic evoked high frequency magnetic oscillations to posterior tibial nerve stimulation. AB - OBJECTIVE: High frequency oscillations (HFOs) evoked by posterior tibial nerve stimulation were recorded using magnetoencephalography (MEG). Time-frequency domain multiple signal classification (TF-MUSIC) algorithm was applied, and the usefulness of this method was demonstrated. METHODS: Ten normal subjects were studied. To localize sources for the HFOs of those somatosensory evoked fields, we applied two kinds of methods: the single moving dipole (SMD) method and the TF MUSIC method. The SMD method was applied after digitally band-pass filtering the somatosensory response with a bandwidth of 500-800 Hz. To estimate the locations of sources with the TF-MUSIC algorithm, we first set the target region on the spectrogram of the somatosensory responses. Then, the procedure described in Section 2.2 was applied with this target region. RESULTS: A clear, isolated region was detected in 6 out of 10 subjects using a time-frequency spectrogram. The averaged distance of the dipole sources between the HFOs and the underlying P37m using the TF-MUSIC algorithm was smaller than using the SMD method. CONCLUSIONS: The TF-MUSIC algorithm is suitable for extracting a target response whose spectrum changes significantly during the observation. PMID- 10479026 TI - N30 and the effect of explorative finger movements: a model of the contribution of the motor cortex to early somatosensory potentials. AB - OBJECTIVES: The source of the N30 potential in the median nerve somatosensory evoked potentials (SEP) has been previously attributed to a pre-central origin (motor cortex or the supplementary motor area, SMA) or a post-central located generator (somatosensory cortex). This attribution was made from results of lesion studies, the behavior of the potential under pathological conditions, and dipole source localization within spherical volume conductor models. METHODS: The present study applied dipole source localization and current density reconstruction within individual realistically shaped head models to median nerve SEPs obtained during explorative finger movements. RESULTS: The SEPs associated with movement of the stimulated hand showed a minor reduction of the N20 amplitude and a markedly reduced amplitude for the frontal N30 and parietal P27, exhibiting a residual frontal negativity around 25 ms. The brain-stem P14 remained unchanged. Mapping of the different SEPs (movement of the non-stimulated hand minus movement of the stimulated hand) showed a bipolar field pattern with a maximum around 30 ms post-stimulus. In eight out of ten normal subjects, both the N30 and the gN30 (subtraction data) sources resided within the pre-central gyrus, more medially than the post-centrally located N20. Two subjects, in contrast, showed rather post-centrally localized sources in this time range. A model of the cortical SEP sources is introduced, explaining the data with respect to previously described findings of dipole localization, and from lesion studies and the alterations seen in motor diseases. CONCLUSIONS: The results provide evidence for a pre-central N30 generator, predominantly tangentially oriented, located within the motor cortex, while no sources were detected elsewhere. It is suggested that the mechanisms underlying the 'gating' effect during explorative finger movements in the 30 ms time range predominantly arise in the motor cortex. PMID- 10479028 TI - The relation between asymmetry and amplitude of the P300 field in schizophrenia. AB - OBJECTIVES: A general reduction of auditory P300 amplitude is one of the most consistently replicated observations in schizophrenic patients. However, a left hemisphere amplitude reduction is discussed controversely. METHODS: Encouraged by this controversy we analysed P300 data of 53 controls and 60 patients of the schizophrenia spectrum (ICD-10: F20, F23, F25). RESULTS: Our main result was the finding of the correlation between general amplitude and hemispheric asymmetry in the patient group: low general amplitudes were assigned to a left hemispheric amplitude reduction, high amplitudes to a right hemispheric reduction. Based on this result, we grouped all subjects according to their hemispheric asymmetry pattern. The control group and the group of schizoaffective patients (F25) with a left hemispheric amplitude reduction showed the inverse asymmetry pattern of P300 amplitude compared to subjects with a right hemispheric amplitude reduction. In contrast, the schizophrenic patients (F20) showed only a difference in P300 amplitudes measured at electrode sites of the left hemisphere. Additionally, we could show, that the amount of the general amplitude reduction in patients was different between the diagnostic groups. CONCLUSIONS: We conclude that a left hemispheric amplitude reduction is not a general feature of schizophrenia but it seems to be pathological when combined with a general amplitude reduction. PMID- 10479027 TI - Mismatch negativity and late auditory evoked potentials in comatose patients. AB - OBJECTIVES: The purpose of this study was to assess the patterns of mismatch negativity (MMN) and N100 component in comatose patients and to evaluate their prognostic value vis-a-vis return of consciousness. METHODS: MMN and auditory (early, middle-latency and late) evoked potentials were recorded in 52 normals and in 128 comatose patients (comas due to neurosurgical and neurological problems). At the time of recording, all patients scored lower than 8 on the Glasgow scale. RESULTS: Visually detected N100 and MMN were confirmed by cross correlation of sub-averages. The MMN was present in 33/128 patients and the N100 component in 84/128. The amplitudes of MMN and N100 waves detected in comatose patients were statistically different from those of normal subjects. By 3 months after the onset of coma, 95 patients had returned to consciousness, most of them with moderate to severe disability. A ratio of 30/33 patients with MMN and 70/84 with N100 had regained consciousness. The presence of a MMN together with a N100 component was more specific (90.9%) than the presence of a N100 component irrespective of MMN (57.6%) in terms of predicting return to consciousness, but its sensitivity was lower (respectively 31.6% for MMN and 73.7% for N100). The mean period that elapsed between the recording of evoked potentials and a return to consciousness was 6.3+/-4 days. MLAEPs were also highly specific, but BAEPs were not. CONCLUSION: MMN and auditory evoked potentials provide a reliable assessment of the functional status of comatose patients. When present, MMN and the N100 differ from those found in normal subjects in terms of latencies and amplitudes. As a predictor of return of consciousness MMN had high specificity and low sensitivity, whereas the N100 had high sensitivity and low specificity. This study demonstrates that the recording of MMN and the auditory N100 can be a very useful aid in the assessment of coma and in predicting whether or not a patient will regain consciousness. PMID- 10479029 TI - Strength-duration properties and their voltage dependence at different sites along the median nerve. AB - OBJECTIVES: There is some evidence that the ease with which ectopic activity can be induced varies systematically along the course of a nerve and is greater at more proximal sites. Recent studies have implicated a non-inactivating threshold conductance, possibly due to persistent Na+ channels, in ectopic activity associated with ischaemia and hyperventilation. This conductance is largely responsible for the voltage dependence of strength-duration time constant (tauSD), and changes in it can explain the time constant changes that occur during hyperventilation and ischaemia. METHODS: To determine whether the strength duration properties of motor axons of the median nerve vary along the course of the nerve, tauSD and rheobase were calculated at wrist, elbow and axilla in 15 healthy subjects, and the relationship of these properties to threshold was assessed using DC polarizing current to change axonal excitability. RESULTS: tauSD was similar at the 3 stimulating sites but increased less at the axilla with depolarizing current. CONCLUSIONS: These data indicate that the greater tendency for ectopic activity to arise from proximal segments of motor axons cannot be explained by differences in the conductances that contribute to tauSD and underlie its dependence on axonal excitability. The findings provide further support for the view that the precise relationship of the stimulating electrodes to the nerve has little effect on tauSD, at least when it is measured in the forearm. PMID- 10479030 TI - Satellite potentials of motor unit action potentials in normal muscles: a new hypothesis for their origin. AB - OBJECTIVE: A satellite potential is a late component of the motor unit action potential (MUAP) that occurs both in pathologic and normal muscle. We investigated the physiological mechanisms responsible for satellite potentials in normal muscle by relating the latencies of MUAP features to the timing of the underlying electrical events. METHODS: We analyzed 21 MUAPs with satellite potentials that had been recorded using a monopolar needle electrode from brachial biceps and tibialis anterior muscles in 10 normal subjects. We estimated the endplate-to-electrode, endplate-to-tendon, and satellite propagation times from the latencies, with respect to the MUAP onset, of the MUAP spike, terminal wave, and satellite. RESULTS: Satellite latencies ranged from 8.8 to 32 ms, too long to be explained by mechanisms involving regenerating axons or atrophic muscle fibers. The spike-to-satellite time intervals approximated either twice the spike-to-terminal-wave interval (17 MUAPs) or twice the terminal-wave latency (4 MUAPs). CONCLUSIONS: These results are consistent with the hypothesis that satellite potentials are due to retrograde propagation in a non-innervated muscle fiber that is connected with an innervated muscle fiber at one of the muscle/tendon junctions. Such a configuration could arise as a result of longitudinal muscle-fiber splitting. PMID- 10479031 TI - Patterned electromyographic activity in the sit-to-stand movement. AB - OBJECTIVES: Postural activity is an essential part of any voluntary movement. In movements involving all body parts such as in the sit-to-stand maneuver (STS), postural activity becomes intermingled with the activity required to perform the actual movement, and is usually difficult to recognize. METHODS: In this paper, we have analyzed the electromyographic activity of muscles of the leg, thigh, trunk and neck in 20 healthy volunteers who performed the STS maneuver. In order to identify the postural and the executional EMG activity, subjects were requested to perform the movement according to 6 predefined experimental conditions. Our hypothesis was that changing the conditions or the strategies of the movement would modify the postural activity, without significantly altering the EMG pattern required for performing the movement itself. RESULTS: Tibialis anterior (TA), abdominal (ABD) and sternocleidomastoid (SCM) were the muscles activated first in the most natural condition for the movement (reference), but their activity and that of the trapezius (TRA) was suppressed or greatly diminished in other conditions. Conversely, lumbar paraspinal (LPS), quadriceps (QUA), and hamstrings (HMS) were activated in a patterned sequence that was invariable in all conditions, and time locked to the moment of take-off from the seat. The soleus (SOL) was the last muscle activated and, together with QUA and HMS, remained active during standing. CONCLUSIONS: We conclude that TA, SOL, ABD, SCM and TRA are involved in preparatory and accompanying postural movements, while LPS, QUA, and HMS are activated in a consistent patterned sequence and can be considered the executional muscles for the STS movement. Recognition of the role of each muscle in the STS movement can contribute to a better understanding of the movement deficits of certain patients, and help to devise adaptive motor strategies in neurorehabilitation. PMID- 10479032 TI - Motor-evoked potentials: unusual findings. AB - OBJECTIVE: The aims of this study were to present rare findings of motor evoked potentials (MEPs) in 3 patients with spastic paraparesis and to show that careful interpretation is indispensable in experiments done with very high intensity stimulation. METHODS: The conduction along several segments of the descending tracts was studied by our previously published method in 3 patients with spastic paraparesis. RESULTS: The threshold for activation of descending tracts was markedly increased in all the patients. In one patient, both transcranial electrical and magnetic cortical stimulation elicited responses with 4 different latencies. They were compatible with the latencies of I1-, D(D1)-, D2- or D3 waves. Very high intensity stimulation elicited D2 waves (activation around the cerebral peduncle) or D3 waves (activation at the foramen magnum level). In the other two patients, unexpectedly, the latency of responses to foramen magnum level stimulation was longer than the cortical latency. Foramen magnum and spinal cord stimulation could not excite the corticospinal tract but activated other slowly conducting descending tracts (about 20 m/s), whereas cortical stimulation activated the corticospinal tract. CONCLUSIONS: The site of activation following cortical stimulation was variable when very high intensity stimulation is used. The descending tracts that contribute to the onset of electromyographic (EMG) responses may not be the same after cortical and spinal stimulation in patients with severely affected corticospinal tract, especially when using very high intensities of stimulation. Such factors complicate the interpretation of EMG responses obtained in patients with severely affected corticospinal tracts. PMID- 10479033 TI - Topographical differences in the developmental profile of central motor conduction time. AB - OBJECTIVES: To study the topographical difference in the developmental profile of the central motor conduction time (CMCT) in upper extremity muscles, electromyographic (EMG) responses to transcranical magnetic stimulation (TMS) were examined in the first dorsal interosseous, extensor carpi radialis (ECR), biceps (BCP), and deltoid (DT) muscles of 25 neurologically normal subjects aged from 2 to 26 years. METHODS: The motor cortex and cervical spinal roots were magnetically stimulated, and CMCT was measured as the onset latency difference between these EMG responses. RESULTS: CMCT in children was shorter in the more proximal muscle of each adjacent muscle pair, despite the tendency of a higher threshold intensity for TMS of the more proximal muscle. This topographical difference tended to be more distinct in younger children, whereas CMCT in adults did not show such a topographical difference. Consequently, the linear decrease in CMCT during maturation was less pronounced in the proximal muscles. CONCLUSIONS: We speculate that direct activation of corticospinal neurons to the more proximal muscles was preferentially produced by TMS in younger children, depending on the relationship between the spatial direction of axons, head circumference, and stimulating coil diameter. PMID- 10479034 TI - Long-term botulinum toxin treatment of cervical dystonia--EMG changes in injected and noninjected muscles. AB - OBJECTIVE: To evaluate changes in quantitative EMG of injected and noninjected sternocleidomastoid muscles following long-term unilateral botulinum toxin treatment of cervical dystonia. METHODS: We investigated 27 patients with cervical dystonia, who received repeated unilateral botulinum toxin injections of the sternocleidomastoid muscle, with quantitative EMG at rest and at maximal voluntary contraction. The patients had on the average 7.1 botulinum toxin treatments and the follow-up period was on the average 31 months (SD 16). RESULTS: After the first treatment, the injected sternocleidomastoid muscles showed a significant decrease in turns/s (mean 45%) and amplitude (mean 52%) at rest, and in amplitude at maximal flexion (mean 24%) and rotation (mean 39%). Except for a reduction in turns/s at rotation (mean 19%) no further reductions in EMG parameters were seen after long-term treatment. The contralateral noninjected sternocleidomastoid muscles showed no significant change in EMG activity after the first BT treatment, but after long-term treatment a significant reduction in turns/s and amplitude at both maximal flexion (turns: mean 28%; amplitude: mean 25%) and rotation (turns/s: mean 32%; amplitude: mean 25%) were seen as compared to pretreatment values. CONCLUSION: The results indicate that there seems to be no cumulative chemodenervation by repeated botulinum toxin injections of sternocleidomastoid muscles measured by quantitative EMG. Contralateral noninjected sternocleidomastoid muscles however, seem to be affected following long-term treatment. The mechanism behind this finding is unknown. PMID- 10479035 TI - Tibial nerve somatosensory evoked potentials during EEG suppression in sevoflurane anaesthesia. AB - OBJECTIVES AND METHODS: Cortical tibial nerve somatosensory evoked potentials (TSEPs) were recorded from 10 subjects in sevoflurane anaesthesia in order to study TSEP during EEG suppression. RESULTS: With a stimulation frequency less than one per second the major component was a positive wave which had maximal amplitude parietally ipsilaterally to stimulus and mean latency of 46.1 ms. It probably corresponds to the P40 wave. It was preceded by a widespread smaller positive wave, which corresponds to the subcortical P30 wave. In two patients a high amplitude negative wave, a couple of milliseconds before the positive wave, and maximal parietally contralateral to stimulus, was seen. All later waves were absent. CONCLUSION: The results are in agreement with our previous results from median nerve SEPs showing that the first cortical response from primary somatosensory cortex is enhanced, and later waves are suppressed. Hence, recording TSEPs during EEG suppression provides a way to record the activity of the primary somatosensory cortex accurately and rapidly due to the very good signal to noise ratio, so that even single responses to stimuli can be seen without averaging. Our results suggest that new cortical generators, which are not recordable awake, may be discovered in some patients. PMID- 10479036 TI - Stability of high-frequency (600 Hz) components in human somatosensory evoked potentials under variation of stimulus rate--evidence for a thalamic origin. AB - The generators of spike-like high-frequency (600 Hz) wavelets superimposed on the primary cortical response (N20) in human median nerve somatosensory evoked potentials (SEP) have been localized anatomically both close to the primary somatosensory hand cortex and in deep axon segments of thalamo-cortical projection neurons. Here, N20 and 600 Hz burst components were functionally dissociated by varying the stimulus rate (1.5, 3, 6, 9 Hz). The N20 source amplitudes were significantly reduced at the higher stimulus rates. In contrast, the source amplitudes of the 600 Hz oscillations remained stable across all stimulus rates. This reflects different source origins, confirming a postsynaptic intracortical generation of the N20 component and provides further evidence for a presynaptic origin of the 600 Hz activity like repetitive neuronal population spikes conducted in deep and superficial segments of thalamo-cortical projection fibers. PMID- 10479037 TI - Latency and amplitude variability in serial median nerve SEP recordings. AB - OBJECTIVES: Intra-individual variability of SEP parameters was investigated by serial SEP recordings. METHODS: Median nerve SEP of 53 normal subjects (mean age 25.5+/-2.7 years) were evaluated. Recordings were repeated 1 week, 2 weeks, and 6 months after the initial recording. RESULTS: Mean values of latencies from right median nerve stimulation were slightly longer (+0.1 ms) as compared to the left. The intra-individual (within stimulation side) variance did not depend on the time interval between recordings. The total within-stimulation-side variance of latencies was 0.056 ms2 for N20 (C3/4' vs. Fz), 0.070 ms2 for N13 (C7 vs. Fz), 0.048 ms2 for N10 (Erb's point vs. Fz), 0.111 ms2 for P9 (C3/4' vs. contralateral forearm), and 0.148 ms2 for P14 (C3/4' vs. contralateral forearm), and was about 3 times smaller than between-side variability with regard to cortical or spinal potentials. Upper limits for latency differences and lower limits for amplitude ratios in repeated recordings, as well as upper and lower limits for between-side latency differences and amplitude ratios were calculated, using F distributions with worst-case assumptions for degrees of freedom and error probability P = 0.05. Upper limits of within-stimulation-side latency differences were 0.55 ms for N20, 0.62 ms for N13, 0.51 ms for N10, 0.78 ms for P9, and 0.90 ms for P14, and corresponding lower limits of amplitude ratios were 0.69, 0.62, 0.66, 0.45, and 0.50, respectively. CONCLUSIONS: Our results demonstrate a small systematic, and a larger random difference between right and left side stimulation, one arising from the peripheral, and the other one from the central pathway proximal to the source of N10. PMID- 10479038 TI - Causal pathways and prevention. PMID- 10479039 TI - Cerebral palsy in Malta 1981 to 1990. AB - Of 55200 Maltese children born in the Mediterranean islands of Malta and Gozo in the birth years 1981 to 1990 inclusive, 134 fitted the case definition of cerebral palsy (CP). Children were allocated to prenatal, perinatal, and postnatal groups according to strict criteria and to various CP syndromes using the Swedish classification. The study was undertaken between 1994 and 1996 and the period prevalence rate was calculated. The control group comprised 134 children who were matched for sex and born immediately before or after the index subjects in the same hospital. Data related to risk factors were extracted from medical records and collected by interview with parents/carers. Unadjusted odds ratios (OR) with 95% confidence intervals for a range of risk factors were calculated. For risk factors with a relatively high prevalence in the population, such as prematurity and low birthweight, ORs were calculated from case-control data. For low-prevalence risk factors, such as breech presentation and multiple pregnancy, ORs were calculated against the whole population data for the 10-year birth cohort. The prevalence rate of CP in the geographically defined, stable population of the Maltese islands was similar to that in the UK and higher than that for Sweden, but the rates for different types of CP were markedly different from most developed countries, with a higher rate of spastic tetraplegias. Risk factors were as expected and similar to those reported from other developed countries. The study shows a strong social gradient with a higher than expected proportion of CP in children of unskilled manual workers. PMID- 10479040 TI - Hemispheric specialization using SPECT and stimulation tasks in children with dysphasia and dystrophia. AB - Developmental dysphasia, a severe childhood learning disorder, is thought to result from problems in hemispheric specialization involving both left and right cerebral hemispheres. Regional cerebral blood flow (rCBF) was measured at rest and during stimulation of both hemispheres independently: dichotic listening for the left, dichaptic palpation for the right. Eight right-handed boys with expressive dysphasia, aged 8 to 12 years, were investigated using SPECT and compared with eight right-handed age-matched boys with Duchenne muscular dystrophy with reading disorders but normal speech. rCBF values at rest were also compared with those of five right-handed age-matched normal boys. In the dichotic task, children with dysphasia differed from children with dystrophia by failure to increase rCBF in the left hemisphere, in Broca's area, but rCBF increased in the right hemisphere, in the region homologous to Broca's area. In the dichaptic task, rCBF increased bilaterally for children with dysphasia whereas in children with dystrophia rCBF increased only in the right hemisphere. At rest the physiological asymmetry was reversed in favor of the right hemisphere in all areas except Broca's area. Surprisingly, the same applied at rest and for all areas in children with dystrophia. These results confirm that functional specialization of both hemispheres is impaired in developmental dysphasia. Moreover, they suggest that learning disabilities associated with Duchenne muscular dystrophy could also be related to abnormal hemispheric specialization. PMID- 10479041 TI - Neurobehavioral evidence for working-memory deficits in school-aged children with histories of prematurity. AB - Cognitive performance in 7- to 9-year-old preterm neonatal intensive-care survivors was compared with that in age-matched control children. Non-verbal memory span, spatial working-memory abilities, planning, set-shifting, and recognition memory for both spatial and patterned stimuli were assessed using the Cambridge Neuropsychological Testing Automated Battery. Relative to children in the control group, neonatal intensive-care unit (NICU) survivors demonstrated 25% more memory errors on the spatial working-memory task. Their use of strategy on this task was similar to a control group of 5-year-olds. Planning times on 'Tower of London' problems were long relative to those of term controls. NICU survivors demonstrated poorer pattern recognition as well as a shorter spatial memory span. The groups did not differ in visual-discrimination learning or in spatial recognition memory. No specific neonatal risk factor accounted for the observed differences, although scores on the Neurobiological Risk Score (NBRS), a composite measure of neonatal risk, did predict several aspects of later task performance. Whether these data reflect a developmental delay in brain maturation in NICU survivors or the presence of a permanent information-processing deficit due to adverse neonatal events must be assessed through continued follow-up. PMID- 10479042 TI - Continuum of reflex excitability in hemiplegia: influence of muscle length and muscular transformation after heel-cord lengthening and immobilization on the pathophysiology of spasticity and clonus. AB - The electromyographic (EMG) responses to tendon taps at the ankle and ensuing muscular twitch forces and temporal parameters were studied at varying angles across the joint range in 18 children, aged 3 to 14.9 years, with congenital hemiparetic cerebral palsy and 22 healthy (control) children, aged 3 to 13.6 years. Those subjects with hemiparesis were community ambulators without assistance. In all subjects, passive muscle stretch caused a waxing of the reflex EMG and twitch force near neutral (with the sole of the foot at right angles to the tibia) and a diminution of these with further dorsiflexion. Twitch times increased with each dorsiflexing increment, being slowest at maximum dorsiflexion and fastest at the resting plantarflexion angle. Heterogeneity of the hemiparetic limb data is evident when compared with data of non-paretic and unaffected limbs, with clear differences in the clonic (fast twitch) as opposed to non-clonic (slow twitch) muscles. In four cases with clinical clonus, clonus frequency was reduced by passive dorsiflexion. Plaster immobilization for 1 month produced clonus which was previously absent in one subject, and caused a fast-twitch phenotype to emerge in two subjects. Follow-up after heel-cord lengthening in one subject showed that clonus frequency diminished from 9 to 3 Hz with slowing and strengthening of muscle-twitch phenotype. Short- and long-term peripheral manipulations appear to regulate neuromuscular excitability according to whether muscles are loaded or unloaded. Although damage to the nervous system provides the setting for reflex excitability, the data suggest that the muscle length (which specifies the joint angle) and the muscle-twitch phenotype of any given limb for any given case appear to dictate the actual speed and strength of reflex muscle-twitch and clonus profiles. This study illustrates how peripheral manipulations of muscles and tendons may alter the expression of what have hitherto been considered as exclusively central phenomena. PMID- 10479043 TI - A long-term follow-up of cognitive, emotional, and behavioural sequelae to Reye syndrome. AB - Eighteen adolescents who had survived Reye syndrome (RS) in early childhood were assessed on cognitive, emotional, and behavioural variables in a second follow-up study tracking this group. Siblings were used as controls. The entire group with RS had survived with no obvious neurological damage at the first follow-up study. Indeed, current findings suggested that long-term cognitive, emotional, and behavioural functioning was comparable to siblings in approximately half of the group with RS. However, two factors were associated with a less favourable outcome. Cognitive, emotional, and behavioural functioning were significantly poorer in the subgroup of survivors whose illness had occurred in the first year of life. In addition, loss of consciousness, although the association with poor outcome was not as noticeable, was also associated with relative deficits on some scales of cognitive ability. Many of these deficits had not been obvious at the first follow-up and the importance of neurodevelopmental factors are considered. Finally, the implications of these findings for research and interventions in RS and other such encephalopathies are discussed. PMID- 10479046 TI - Children's sleep disorders: modern approaches, developmental effects, and children at special risk. PMID- 10479045 TI - Apneustic breathing in children with brainstem damage due to hypoxic-ischemic encephalopathy. AB - To confirm the presence of apneusis in patients with hypoxic-ischemic encephalopathy and to clarify which factors influence their respiratory patterns, polygraphic studies were performed on two patients. Apneusis was clinically suspected in both patients who had severe brainstem damage. In one subject, inputs of vagal afferents from the gastrointestinal tract and the urinary bladder often resulted in extreme tachypnea instead of apneusis. Lung inflation facilitated expiration during inspiratory arrest. Expiration preceded a periodic inhibition of rigospastic discharge in the right biceps muscle. In the other subject, prolonged inspiratory pauses with cyanosis occurred with or without preceding epileptic seizure. Both phenytoin dose reduction and treatment with tandospirone, a serotonin-1A agonist, were effective in improving the respiratory distress in this subject. PMID- 10479044 TI - Health-related quality of life in childhood epilepsy: the results of children's participation in identifying the components. AB - Separate groups of children with epilepsy, recruited from a regional pediatric epilepsy database, and their parents were established to discuss their life with epilepsy. Twenty-nine children (aged between 6 years and 10 years 4 months) and 42 of their parents were placed into nine and 17 groups respectively. The participants provided information about their own perceptions of life with epilepsy. Discussions were taped and textual analysis followed to extract, understand, explain, and categorize the health-related quality of life (HRQL) components. The process enabled us to identify the burdens and concerns of children with epilepsy, and to identify five emerging dimensions: (1) the experience of epilepsy, (2) life fulfillment and time use, (3) social issues, (4) impact of epilepsy, and (5) attribution. Identifying and understanding the components of HRQL is crucial for developing an HRQL scale in childhood epilepsy. In addition, this list of elements can help health professionals improve their services by considering and addressing aspects of the epilepsy experience beyond the traditional issues for children with epilepsy and their families. PMID- 10479047 TI - Polymorphism of HLA-G gene and protein. PMID- 10479048 TI - Evolution of MHC-G in primates: a different kind of molecule for each group of species. AB - When MHC-G molecules in primates (New World and Old World monkeys, Anthropoids and humans) were compared phylogenetically, very different evolutionary patterns within each species were found; their molecules did not have a straight forward and linear development throughout the postulated evolutionary pathway of primates. The earlier New World monkeys (South America) had relatively more alleles and the polymorphism was placed in the T-cell receptor (TcR), NK receptors and antigen binding sites; MHC-G probably works as a classical class I presenting molecule in these monkeys. MHC-G intron 2 from New World monkeys does not show the typical 23 bp deletion found in all other more recent primate species. Thus, it is possible that MHC-G molecules in New World monkeys belong to a different lineage than the MHC from higher primates. Another early lineage, Eurasian Old World monkeys, shows stop codons at exon 3: MHC-G proteins lacking the alpha2 domain may functionally suffice or otherwise reading-through stop codon translational mechanisms may exist, as shown for other genes. Orangutans show lower (but significant) polymorphism than New World monkeys at NK, TcR and antigen binding regions; gorilla and chimpanzee show very low polymorphism. Humans only show three different HLA-G proteins with changes not affecting NK, TcR or antigen binding sites. It is observed that the more exposed the mother to allogeneic fetuses (polygamy), the less polymorphic HLA-G is observed within a given species. The data are concordant with the postulated immune inhibitory function for MHC-G in Old World monkeys, anthropoids and humans both at placental and inflammatory level. PMID- 10479049 TI - Human leucocyte antigen-G and its recognition by natural killer cells. PMID- 10479050 TI - Peptide presentation and NK inhibition by HLA-G. AB - The expression of the nonclassical MHC class Ib molecule HLA-G is nearly exclusively restricted to the feto-maternal interface during pregnancy. There it probably serves the same physiological functions already known for classical MHC class I molecules; these include peptide presentation, natural killer cell (NK) inhibition and probably also T cell restriction. In this study a comparison between HLA-G and HLA-A2 as far as the amount and complexity of bound peptides is concerned revealed no significant differences. The peptide motif of HLA-G, as determined by analysis of naturally eluted peptides allows the construction of a peptide library that is efficient in binding to HLA-G and thereby confirms the rules of peptide binding to this nonclassical MHC class I molecule. In addition, we demonstrate that the inhibition of NK cells by HLA-G varies remarkably among the NK repertoires of different donors. The function of HLA-G as a survival factor in the development of the fetus during pregnancy is discussed in detail. PMID- 10479051 TI - P49, a putative HLA-G1 specific inhibitory NK receptor belonging to the immunoglobulin Superfamily. AB - NK cells display several killer inhibitory receptors (KIRs) specific for different alleles of major histocompatibility complex (MHC) class I molecules. A family of KIRs are represented by type I transmembrane proteins belonging to the Immunoglobulin Superfamily (Ig-SF). In the present study we describe a cDNA, termed cl.15.212, that encodes for a type I transmembrane protein displaying approximately 50% sequence homology with other Ig-SF members. The protein encoded by cl.15.212 (termed p49 according to its apparent molecular weight of 49 kDa) is characterized by two extracellular Ig-like domains, a 115-amino acid cytoplasmic tail containing a single immuno-receptor tyrosine-based inhibitory motif (ITIM) typical of KIR. Different from the other KIRs, the cl.15.212 transcript is expressed by all NK cells and by a fraction of T-cell clones expressing KIR. To determine the specificity of the cl.15.212-encoded receptor, we generated a chimeric protein, formed by the ectodomain of p49 and the Fc portion of human IgG1 (p49-Fc). Soluble molecules bound efficiently to LCL721.221 (221) cells transfected with HLA-G1, -A3, -B46 alleles and weakly to the -B7 allele. On the other hand, they did not bind to 221 cells either untransfected or transfected with HLA-A2, -B51, -Cw3, or-Cw4. PMID- 10479052 TI - NK cell mediated recognition of HLA class Ib molecules: role of CD94/NKG2 receptors. PMID- 10479053 TI - Role of HLA-G versus HLA-E on NK function: HLA-G is able to inhibit NK cytolysis by itself. AB - Recent studies have shown that endogenous HLA-E molecules are stabilized on the cell surface upon the expression of HLA-G which contains within its leader sequence, a nonapeptide capable of binding with the HLA-E/beta2m complex. Since HLA-E was found to be the major ligand for the CD94/NKG2A inhibitory receptor, we determined the role of HLA-G versus HLA-E on NK lysis inhibition. We showed that K562 cells transfected with HLA-G1 cDNA are protected from NK lysis by direct interaction between HLA-G1 and killing inhibitory receptor(s). This NK lysis inhibition is not dependent on HLA-E expression, since no HLA-E protein was detected on K562 cells; HLA-G1 is therefore able to inhibit NK lysis by itself. PMID- 10479054 TI - HLA-G expression in human melanoma cells: protection from NK cytolysis. AB - Expression of the non-classical HLA-G class I antigen is physiologically restricted to a limited number of tissues including trophoblasts, and is thought to play a role in establishing tolerance of the fetus by the maternal immune system. We investigated whether ectopic expression of HLA-G could also be detected in tumor cells and confer them the ability to escape immune cytotoxic responses. High levels of all alternatively spliced HLA-G transcripts could be detected in melanoma cells by RT-PCR. Analysis of biopsies from a melanoma patient revealed a higher HLA-G transcription level in skin metastasis as compared to healthy skin, while specific amplification of the HLA-G5 transcript was only observable in the tumor. HLA-G protein expression could also be detected in two melanoma cell lines. HLA-G-positive tumors inhibit cytotoxic lysis by the NK cell line YT2C2-PR. This inhibition is not observed with B-EBV cell lines bearing matched class I specificities, and is thought to occur through interaction of HLA-G with inhibitory receptors that are distinct from known KIRs interacting with HLA-E or classical class I molecules. Together, these results confirm that HLA-G expression at the surface of tumor cells can participate in the evasion of antitumoral immune responses and favor tumor progression. PMID- 10479055 TI - Cell surface detection of HLA-E gene products with a specific monoclonal antibody. AB - An HLA-E-specific monoclonal antibody (mAb) was obtained by immunization of human beta2-microglobulin transgenic mice (M-TGM) with spleen cells from double transgenic mice expressing HLA-E molecules (EM-TGM). This mAb, designated V16, specifically recognizes in flow cytometry analysis the HLA-E expressing mouse cells, whereas it does not bind to mouse cells expressing various HLA class I molecules (HLA-A2, -A3, -A11, -A26, A29, -B7, -B27, -Cw3, -Cw7, and HLA-G). V16 mAb binds efficiently to human EBV-infected B lymphocytes, PHA blasts and PBL, thus establishing the surface expression of HLA-E in vivo on these cells. PMID- 10479056 TI - HLA-G inhibits the allogeneic proliferative response. AB - HLA-G is a non-classical MHC class I molecule expressed at the feto/maternal interface where it plays a role in materno-fetal tolerance by inhibiting NK cells. Expression of killing inhibitory receptors capable of interacting with HLA G on T lymphocytes led us to hypothesize that HLA-G molecules could also modulate T cell responses, analyzed here in the context of the allogeneic proliferative response. Using LCL-HLA-G transfectants as stimulators of T cells present among peripheral mononuclear cells and K562-HLA-G1 transfectants as inhibitors in a classical mixed lymphocyte reaction, we showed that HLA-G is able to inhibit T cell allo-proliferation. These findings provide new insight into the role of HLA G in preventing allograft rejection. PMID- 10479057 TI - Molecular mechanisms controlling constitutive and IFN-gamma-inducible HLA-G expression in various cell types. AB - HLA-G molecule is thought to play a major role in down-regulating the maternal immune response by inhibiting NK and T cell cytolytic activities. We examined the molecular regulatory mechanisms that may control the restricted expression pattern of the HLA-G gene. We first analyzed protein interactions between nuclear extracts from the HLA-G-positive JEG-3 choriocarcinoma and the HLA-G-negative NK like YT2C2 cell lines to a 244 bp regulatory element located 1.2 kb from the HLA G gene, previously shown to direct HLA-G expression in transgenic mouse placenta. This allowed characterization of cell-specific DNA-protein interactions that could account for differential cell-specific expression of the HLA-G gene. In particular two DNA-protein complexes were exclusively observed in YT2C2, suggesting that this HLA-G regulatory element is a target for putative cell specific repressor factors. We further mapped nuclear factor binding sites to a 70 bp fragment in the upstream region of the regulatory element. We then investigated the effect of IFN-gamma on HLA-G gene expression. HLA-G cell surface expression was enhanced by IFN-gamma treatment in JEG-3 and U937 cell lines and peripheral blood monocytes while no effect was observed in tera-2 teratocarcinoma cell line. HLA-G transcriptional activity was increased only in JEG-3 and U937 cell lines. Activity of the 1.4-kb HLA-G promoter region was unchanged after IFN gamma treatment in JEG-3 and Tera-2. These results suggest that both post transcriptional and transcriptional mechanisms implicating IFN-responsive regulatory sequences outside the 1.4 kb-region are involved in IFN-gamma gene activation of the HLA-G gene. PMID- 10479058 TI - Primary cultured human thymic epithelial cells express both membrane-bound and soluble HLA-G translated products. AB - This report demonstrates that both membrane-bound and soluble HLA-G isoforms are present in primary cultured human thymic epithelial cells (TEC). HLA-G transcriptional isoforms have been detected by RT-PCR, using different sets of HLA-G specific primers. A flow cytometry analysis, using two anti-HLA-G mAbs, namely 87G and BFL.1, revealed the presence of HLA-G translated products at the cell surface of a subpopulation of TEC. Finally, it was shown that HLA-G soluble forms were secreted in TEC culture supernatant, using a sandwich ELISA with BFL.1 and W6/32 mAbs. These results confirm and extent those previously described showing that HLA-G expressing cells were detectable by immunohistochemistry in thymic medullary epithelial cells. PMID- 10479060 TI - Viral immunoevasive strategies and trophoblast class I major histocompatibility complex antigens. PMID- 10479059 TI - Human leucocyte antigen-G and reproduction. PMID- 10479061 TI - HLA-G transgenic mice. AB - We have generated a number of transgenic mice using DNA segments derived from the HLA-G gene. Using these mice we have examined the pattern of expression dictated by HLA-G promoter elements in mice and shown that HLA-G functions both as a restriction element and a transplantation antigen recognized by murine T cells. In addition, we have shown that trophoblast cells expressing H-2Kb under the control of HLA-G promoter elements affect maternal T cell phenotype and responsiveness during pregnancy. Using these same HLA-G/H-2Kb transgenic mice we have shown that trophoblast cells, expressing an inducible enzyme that degrades tryptophan, protects allogeneic conceptus expressing paternally-inherited transgenes from attack by maternal T cells that leads to fetal rejection. PMID- 10479062 TI - Looking for HLA-G expression in human tumours. AB - Tumour and virus infected cells escape CTLs responses by losing some or all HLA class I molecules. However the NK escape mechanism that uses the HLA-A, -B, and C tumour deficient variants is unknown. To determine whether HLA-G is expressed on tumour cells and thus favours tumour escape by abolishing NK lysis, we studied HLA-G in a large panel of human tumour tissues and human tumour cell lines of different origin that were previously characterized for HLA-A, -B, and -C expression. We studied HLA-G mRNA transcripts using RT-PCR, and HLA-G1 expression by FACS and immunohistochemical techniques. We found several mRNA transcripts of HLA-G isoforms in most of the samples studied. However, we detected no cell surface expression of HLA-G1 using two specific monoclonal antibodies (mAbs) (87G and 01G). We cannot, however, exclude the possibility that some isoforms other than HLA-G1 may be expressed in some tumours. PMID- 10479063 TI - Expression of synaptic proteins in the developing rat cerebellum following ionizing radiation. AB - Various proteins regulating neurotransmission release and synaptic vesicle exocytosis have been implicated in axonal elongation and synaptic maturation. In the present study, immunohistochemistry to the presynaptic membrane proteins syntaxin-I and synaptosomal-associated protein of 25 kDa (SNAP-25) synaptic vesicle-associated proteins synaptophysin and synapsin-I and the neuronal maturation and axonal growth-related protein GAP-43, has been carried out in the normal developing cerebellum and following a single dose of ionizing radiation (2 Gy gamma-rays) at postnatal day 1. Our aim has been to learn about the morphological and possible functional modalities that occur during the progression of neuronal connectivity in normal and abnormal development. Expression of all these proteins is associated with the arrival of afferents in the subcortical white matter and with the maturation of the internal granule cell layer and molecular layer during normal development. In addition, SNAP-25 and GAP 43 are strongly expressed in granule cells of the external granule cell layer, thus suggesting that these proteins are involved in cell elongation of granule cells. Apoptosis appears at 3 h and peaks at 6 h following ionizing radiation. Radiation-induced apoptosis in the external granule cell layer produces a transient decrease in the expression of SNAP-25 and GAP-43 in the external granule cell layer. The external granule cell layer recovers at 48 h and external granule cells of proliferating cells also express SNAP-25 and GAP-43, thus indicating that proliferating cells in this layer are equipped with proteins involved in cell elongation. Furthermore, expression of synaptophysin, synapsin I, syntaxin-I and SNAP-25 is the same in the cerebellum of irradiated and normal rats from this time to adulthood (3 months). These results point to the likelihood that recovery of the cerebellar cortex occurs following a single exposure of ionizing radiation during postnatal development. PMID- 10479064 TI - Distribution of BDNF, NT-3 and NT-4 in the developing auditory brainstem. AB - This study describes the developmental expression of three neurotrophins, brain derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3) and neurotrophin (NT-4) in the rat auditory brain-stem using immunohistochemistry. At postnatal day 0 (PND 0), neurotrophins expression was virtually absent from all auditory nuclei in the brainstem, even though some positive neurons were observed in the mesencephalic trigeminal nucleus at this age. However, BDNF, NT-3 and NT-4 positive neurons were observed in most brainstem auditory nuclei by PND 6. At the following stages, there was a general increase in the intensity of the neurotrophins immunoreactivity and BDNF labeling was particularly prominent in most cochlear nucleus neurons. A differential pattern of staining emerged in cochlear nucleus subdivisions, with more intense staining present in the ventral part. The superior olivary complex nuclei followed a similar pattern of BDNF staining compared to the cochlear nucleus. In the adult, BDNF heavily labeled most neurons of the superior olivary nuclei and moderately labeled neurons of the inferior colliculus (IC). NT-3 and NT-4 showed a similar pattern of staining in most auditory brainstem nuclei. The first staining was observed by PND 6 in some neuronal cell bodies. NT-3 and NT-4 immunoreactivity increased in the following stages and in the adult moderate labelings were observed in most neurons of the cochlear nucleus, the superior olivary nuclei and the IC. These results show that neurotrophins are expressed 1 week before the onset of hearing and the increase of their expressions correlate with the appearance of sound-evoked activity in the system. The temporal distribution of neurotrophins does not correlate with neuronal birth, axonal outgrowth or the formation of connection in the auditory structures, suggesting a role primarily in the maintenance and/ or modulation of postnatal and adult functions. PMID- 10479065 TI - Differential effects of prepubertal chronic oestrogen treatment on the synthesis of noradrenaline in uterine myometrial and perivascular sympathetic nerves. AB - Previous studies have shown that chronic administration of oestrogen to prepubertal rats reduces the total content of noradrenaline in the uterine horn, abolishes myometrial noradrenergic innervation and reduces noradrenaline fluorescence intensity of intrauterine perivascular nerve fibres. The mechanisms underlying these changes are not known. In the present study we have analysed the effects of prepubertal chronic oestrogen treatment on the synthesis of noradrenaline in the rat uterine sympathetic nerves using biochemical and immunohistochemical approaches. Tyrosine hydroxylase activity was evaluated biochemically, by measuring the in vivo accumulation of dihydroxyphenylalanine (DOPA) in the presence of a DOPA-decarboxylase inhibitor. In addition, nerve fibres were visualised immunohistochemically using antibodies against tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DbetaH) and the general marker protein gene product 9.5 (PGP 9.5). After chronic oestrogen treatment, the total content of noradrenaline of the uterine horn was reduced, whereas the total content of DOPA was increased. In controls, TH-immunoreactive, DbetaH immunoreactive and PGP 9.5-immunoreactive nerve fibres were distributed in both the circular and longitudinal myometrial layers and in the blood vessels of the intran-myometrial region. After chronic oestrogen treatment the only fibres recognised by the three antibodies were those associated with the blood vessels, but no myometrial-associated fibres could be recognised. These results suggest that noradrenaline synthesis is selectively reduced in myometrial-associated uterine sympathetic nerves, but is preserved in perivascular sympathetic nerves. The increased DOPA levels measured after chronic exposure to oestrogen was interpreted as the consequence of the substantial increase in size and number of blood vessels observed in the uterus of oestrogen-treated animals. A possible neurodegenerative effect of oestrogen on myometrial sympathetic fibres is discussed. PMID- 10479066 TI - Plasticity of 5-hydroxytryptamine(1B) receptors during postnatal development in the rat visual cortex. AB - The distribution of 5-hydroxytryptamine1A and 5-hydroxytryptamine1B receptors in the visual cortex was studied by quantitative autoradiography during postnatal development. Overall, receptor densities increased throughout development, but exhibited regional rearrangements, particularly in the case of 5 hydroxytryptamine1B receptors. Neonatal treatment with 5,7-dihydroxytryptamine, which causes selective degeneration of serotoninergic neurons, had no effect on the density of 5-hydroxytryptamine1A receptors in the visual cortex. However, a transient increase in 5-hydroxytryptamine1B at postnatal days 10-12 was observed after this treatment, suggesting a regulation of postsynaptic receptors. Neonatal enucleation resulted in a marked increase in 5-hydroxytryptamine1B binding sites in all layers of the visual cortex by P16, whereas it had no effect upon 5 hydroxytryptamine1A binding sites. These results show that both receptor subtypes do not exhibit striking transient features in the visual cortex during postnatal development, but rather undergo discrete reorganizations. 5-Hydroxytryptamine1B receptors show changes in density after either neonatal degeneration of serotoninergic neurons or enucleation, indicating that the serotoninergic system involving this receptor subtype can exhibit some postnatal plasticity in the visual cortex. PMID- 10479067 TI - Cellular localization of estrogen receptors on neurones in various regions of cultured rat CNS: coexistence with cholinergic and galanin receptors. AB - Autoradiographic studies have shown that many neurones in explant cultures of rat neocortex, hippocampus, preoptic area and spinal cord express binding sites for [3H]-estradiol which are distributed over the cell bodies and primary processes. By means of immunohistochemistry, it was observed that neurones were labelled by monoclonal antibodies against estrogen alpha-receptors and a polyclonal antibody against estrogen beta-receptors. Immunoreactivity was distributed over the soma and primary processes of the cells, the nuclei being more intensely stained. Double-immunostaining revealed a colocalization of estrogen alpha- and beta receptors on approximately half of the neurones in cultures from neocortex and hippocampus whereas in cultures from preoptic area and spinal cord only few cells were double-stained. On many neurones, a coexistence of estrogen receptors and cholinergic muscarinic or nicotinic sites was found. Furthermore, combined autoradiographic and immunohistochemical studies have shown a colocalization of receptors for estrogen and the neuropeptide [125I]-galanin. The coexistence of estrogen and cholinergic sites as well as of estrogen and galanin receptors on the same neurones are discussed with respect to neurodegenerative events such as Alzheimer's disease. PMID- 10479068 TI - Developmental expression of calretinin-immunoreactivity in the thalamic eminence of the fetal mouse. AB - An investigation of the developmental expression of calretinin immunoreactivity and mRNA expression was carried out in the developing mouse diencephalon. Attention was focused on the thalamic eminence, which is a prominent structure previously described in the thalamus of the fetal mammalian diencephalon and adult lower vertebrates. Calretinin-positive staining was first observed in the thalamic eminence beginning at embryonic day 11. In situ hybridization histochemistry confirmed the presence of calretinin mRNA in the thalamic eminence. During subsequent embryonic development calretinin expression was very intense in neurons in the thalamic eminence though embryonic day 17, and thereafter, was increasingly difficult to distinguish. By postnatal day 0 the thalamic eminence was no longer discernable. Additional neurons within the murine diencephalon also expressed calretinin positive cell bodies and, or neuronal processes, including the stria medullaris, the habenular commissure and the paraventricular thalamic nucleus. It is possible that the thalamic eminence may form during development in order to act as an organizing center for the diencephalon. PMID- 10479069 TI - Fetal human cortical neurons grown in culture: morphological differentiation, biochemical correlates and development of electrical activity. AB - Cultured fetal human cortical neurons derived from second trimester human fetal cortex were analyzed with regard to their morphological differentiation and expression of cell-specific markers. The culture method was adapted from standardized protocols originally developed for the isolation and culture of rodent oligodendrocytes and astrocytes. This technique takes advantage of the different adhesive properties and stratification of central nervous system cells in vitro. Under these culture conditions fetal human cortical neurons underwent morphological differentiation, expressed neuron-specific markers and voltage- and ligand-gated ion channels. Highly enriched cultures of microglia and astrocytes generated from the same starting material also expressed cell-type specific markers. These cultures serve as a valuable tool for the establishment of normative data and as experimental models for neurodevelopmental and neurodegenerative studies. PMID- 10479070 TI - Fluoride-induced depletion of polyphosphoinositides in rat brain cortical slices: a rationale for the inhibitory effects on phospholipase C. AB - Fluoride, which is used commonly as a pharmacological tool to activate phosphoinositide-phospholipase C coupled to the heterotrymeric Gq/11 proteins, inhibited the phosphorylation of phosphatidylinositol (PtdIns) to polyphosphoinositides (PtdIns4P and PtdIns4,5P2) in membranes from rat brain cortex. Fluoride enhanced basal production of 3H-inositol phosphates in membranes prepared from brain cortical slices that had been prelabeled with [3H]inositol, but inhibited the stimulation elicited by carbachol in the presence of GTPgammaS. However in both cases fluoride depleted [3H]PtdIns4P content by 95%. The inhibitory effects of fluoride on the release of 3H-inositol phosphates in slices were not apparent in a pulse [3H]inositol-labeling strategy, but became dramatic in a continuous labeling protocol, particularly at long incubation times. Prelabeling slices with [3H]inositol in the presence of fluoride precluded polyphosphoinositide labeling, and eliminated phospholipase C responsiveness to carbachol under normal or depolarizing conditions, and to the calcium ionophore ionomycin. The lack of response of 3H-polyphosphoinositide-depleted slices to phospholipase C stimuli was not due to fluoride poisoning, unaccessibility of the [3H]inositol label to phospholipase C or desensitization of Gq/11, as the effect of carbachol and GTPgammaS was restored, in the presence of ATP, in membranes prepared from slices that had been labeled in the presence of fluoride. In conclusion, our data show that fluoride, at a concentration similar to that used to stimulate directly Gq/11-coupled phospholipase C, effectively blocks the synthesis of phospholipase C substrates from PtdIns. PMID- 10479071 TI - Repeated seizure-associated long-lasting changes of N-methyl-D-aspartate receptor properties in the developing rat brain. AB - Glutamate NMDA receptor has been implicated in brain developmental processes as well as in excitotoxicity and seizure mediation. A previous study has shown that an acute episode of seizures for 30 min in rats altered NMDA receptor characteristics, mainly in the very immature animal. In order to assess whether receptor modifications may also account for long-lasting cerebral disabilities, medium- and long-term consequences of repeated seizures in developing rats on brain NMDA receptor properties were investigated. Seizures were induced once a day for 3 consecutive days, either from post-natal day 5 (P5) to P7 or from P15 to P17. NMDA receptors were then analysed at P15, P25 and P60 (adulthood) by measuring specific binding of [3H]MK-801 on brain membrane preparations. In addition, allosteric modulation of NMDA receptors by exogenous glutamate and glycine was investigated. Seizures from P5 to P7 led to a 22% increase in the density of [3H]MK-801 binding sites measured at P15, but did not affect NMDA receptor density or affinity at P25 or P60. P15-P17 seizures led to a 21% decrease in the density of binding sites and to a 33% decrease in receptor dissociation constant at P25, while they were without effect at P60. Moreover, P5 P7 and P15-P17 seizures were both associated with a suppression of the glutamate/glycine-induced receptor activation at P60. These modifications might account for long-term alterations in cerebral excitability or plasticity after early convulsive disorders, with regards to altered cognitive capacities, epileptogenesis and brain susceptibility to recurrent seizures. PMID- 10479072 TI - Neuroreceptor mRNA expression in the rat mesenteric artery develops independently of innervation. AB - Studies in autonomic targets have shown that nerves may be required for the development and maintenance of postsynaptic receptor populations. We have examined this relationship in the rat mesenteric artery, assessing mRNA expression levels for a range of neuroreceptors after neonatal sympathectomy, using 6-hydroxydopamine or antisera directed against nerve growth factor, and sensory denervation, using capsaicin. Total RNA was extracted from 28 day old rats and reverse transcription-polymerase chain reaction was performed, using primers specific for the alpha1(A,B,D)- and alpha2(A,B,C)-adrenergic, neurokinin (NKI-NK3), muscarinic (M1-M5) and P2X purinergic (P2x1-7) receptor families. Results showed no decreases in mRNA expression of any of the specific receptor subtypes after either sympathetic or sensory denervation. Small increases in mRNA expression were detected following sensory denervation for some of the receptor subtypes. We conclude that neither sympathetic nor sensory nerves are mandatory for the expression of mRNA of a range of neuroreceptors in the mesenteric vascular bed of the rat. PMID- 10479073 TI - Localization of urokinase-type plasminogen activator, its receptor, and inhibitors in mouse forebrain during postnatal development. AB - Proteolytic enzymes are postulated to play a role in cell migration and synapse organization during brain development. Among these, urokinase-type plasminogen activator (uPA) has been studied in neoplastic and cultured brain cells extensively. We hypothesized that uPA, its receptor, and its inhibitors would be expressed in immature glial and neuronal cells in postnatal mouse forebrain. Immature cortical neurons were immunoreactive for uPA, its receptor, and its substrate plasminogen peaking at the end of postnatal week two, consistent with the postulated role in synaptogenesis. Immunoreactivity for uPA receptor was also observed on astroglial cells in vitro. Neither it nor uPA were convincingly detected in subventricular zone precursor cells, immature white matter or pre labeled immature cells that had been transplanted into brain. Plasminogen activator inhibitor type 1 immunoreactivity was observed on endothelia up to 12 days age, and type 2 was observed to surround immature cells. We conclude, based on the spatial and temporal distribution of immunoreactivity, that uPA and its receptor may be relatively more important for synaptogenesis, remodeling, and reactive processes than for cell migration in developing mouse brain. PMID- 10479074 TI - Mesencephalic innervation of the vibrissal follicle-sinus complex in the mouse embryo. AB - Peripheral projections of neurones whose cell bodies lie in the mesencephalic nucleus of the fifth cranial nerve, situated between the central grey and mesencephalic reticular formation, were studied in mouse embryos aged between day 9 and 15 and in postnatal day 1 mice. Nonspecific neural antibody staining allowed visualisation of the developing cranial nerves, in particular the descending mesencephalic tract. This facilitated successful dissection of the descending mesencephalic tract and trigeminal ganglion in the heads of fresh mouse embryos and postnatal mice. The fluorescent dye, 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate (Dil), was injected into the descending mesencephalic tract in mouse embryos aged 12.5, 13.5 and 15 days of gestation and also into postnatal day 1 mice. Following a period of incubation, 100 microm sections were viewed under visible light and episcopic fluorescence. Mesencephalic neurones were observed to pass superiorly over the trigeminal ganglion and enter the maxillary division to innervate vibrissal follicle-sinus complexes, whilst none was observed innervating mandibular and maxillary intraoral structures. There was no fluorescent labelling in non-Dil injected control specimens. Using a highly specific neuronal tracer, this study shows that mesencephalic neurones in the periphery project exclusively to follicle sinus complexes in the developing mouse embryo and remain at least until postnatal day 1. These observations, contrary to those made in other animals, indicate a species specificity of mesencephalic peripheral projections. PMID- 10479075 TI - Pathology, isolation, and preliminary molecular characterization of a novel iridovirus from tiger salamanders in Saskatchewan. AB - All iridovirus was confirmed to be the cause of an epizootic in larval and adult tiger salamanders (Ambystoma tigrinum diaboli) from four separate ponds in southern Saskatchewan (Canada) during the summer of 1997. This organism also is suspected, based on electron microscopic findings, to be the cause of mortality of larval tiger salamanders in a pond over 200 km to the north during the same year. Salamanders developed a generalized viremia which resulted in various lesions including: necrotizing, vesicular and ulcerative dermatitis; gastrointestinal ulceration; and necrosis of hepatic, splenic, renal, lymphoid, and hematopoietic tissues. In cells associated with these lesions, large lightly basophilic cytoplasmic inclusions and vacuolated nuclei with marginated chromatin were consistently found. Virus was isolated from tissue homogenates of infected salamanders following inoculation of epithelioma papilloma cyprini (EPC) cells. The virus, provisionally designated Regina ranavirus (RRV), was initially identified as an iridovirus by electron microscopy. Subsequent molecular characterization, including partial sequence analysis of the major capsid protein (MCP) gene, confirmed this assignment and established that RRV was a ranavirus distinct from frog virus 3 (FV3) and other members of the genus Ranavirus. Intraperitoneal inoculation of 5 x 10(6.23) TCID50 of the field isolate caused mortality in inoculated salamanders at 13 days post infection. Field, clinical, and molecular studies jointly suggest that the etiological agent of recent salamander mortalities is a highly infectious novel ranavirus. PMID- 10479076 TI - Humoral response and protection from experimental challenge following vaccination of raccoon pups with a modified-live canine distemper virus vaccine. AB - Eight 8-wk-old raccoon pups (Procyon lotor) with maternal canine distemper virus (CDV) neutralizing antibodies (NAb) and 24 8-wk-old seronegative pups were administered a commercial modified-live CDV vaccine (Galaxy, D, Solvay Animal Health, Inc., Kitchener, Ontario, Canada). All 24 seronegative raccoons had detectable serum CDV NAb titers 14 days after the initial dose. Titers rose to maximum levels 4 wk post-vaccination. Mean titers for groups of vaccinated seronegative pups were maintained between 1:256 and 1:2,048 for the remainder of the 3 mo observation period. Geometric means of the serum CDV NAb titer of eight seronegative pups given a single vaccine dose at 8 wk of age did not differ significantly from those of eight pups that were given serial doses at 8, 12, and 16 wk of age, or from those of eight pups vaccinated once at 16 wk of age. Seven unvaccinated 8-wk-old raccoon pups used as controls remained seronegative throughout the trial. Seven out of eight 8-wk-old pups with maternal antibodies, vaccinated at 8, 12, and 16 wk of age, failed to develop a rise in their CDV NAb titers until at least 18 wk of age, 2 wk after the third vaccination. Titers in eight unvaccinated raccoons with maternal antibodies declined steadily to undetectable levels at 20 wk of age. A half-life of 10.55 days was calculated for maternally-derived CDV NAb in raccoon pups. Sixteen vaccinated raccoons were protected from clinical disease following experimental oronasal challenge with a virulent raccoon strain of CDV, 13 to 23 wk after vaccination. Serum CDV NAb titers at the time of challenge ranged from 1:12 to 1:384 and increased during the period of observation. Three of four unvaccinated seronegative raccoons used as controls failed to mount any detectable CDV NAb and were euthanatized after developing clinical signs of canine distemper 26, 29, and 30 days post-challenge (PC). Necropsies confirmed the diagnosis. The fourth control raccoon exhibited transient equivocal clinical signs, mounted a sluggish humoral response, but was clinically normal when euthanatized 42 days PC. In this raccoon, there was focal non-suppurative encephalitis with intranuclear inclusion bodies typical of CDV infection. PMID- 10479077 TI - Antibodies against Pasteurella multocida in snow geese in the western Arctic. AB - To determine if lesser snow geese (Chen caerulescens caerulescens) are a potential reservoir for the Pasteurella multocida bacterium that causes avian cholera, serum samples and/or pharyngeal swabs were collected from > 3,400 adult geese breeding on Wrangel Island (Russia) and Banks Island (Canada) during 1993 1996. Pharyngeal swab sampling rarely (> 0.1%) detected birds that were exposed to P. multocida in these populations. Geese with serum antibody levels indicating recent infection with P. multocida were found at both breeding colonies. Prevalence of seropositive birds was 3.5% at Wrangel Island, an area that has no recorded history of avian cholera epizootics. Prevalence of seropositive birds was 2.8% at Banks Island in 1994, but increased to 8.2% during 1995 and 1996 when an estimated 40,000-60,000 snow geese were infected. Approximately 50% of the infected birds died during the epizootic and a portion of the surviving birds may have become carriers of the disease. This pattern of prevalence indicated that enzootic levels of infection with P. multocida occurred at both breeding colonies. When no avian cholera epizootics occurred (Wrangel Island, Banks Island in 1994), female snow geese (4.7%) had higher antibody prevalence than males (2.0%). PMID- 10479078 TI - Development of a model of natural infection with Mycobacterium bovis in white tailed deer. AB - The objective of this study was to develop a suitable experimental model of natural Mycobacterium bovis infection in white-tailed deer (Odocoileus virginianus), describe the distribution and character of tuberculous lesions, and to examine possible routes of disease transmission. In October 1997, 10 mature female white-tailed deer were inoculated by intratonsilar instillation of 2 x 10(3) (low dose) or 2 x 10(5) (high dose) colony forming units (CFU) of M. bovis. In January 1998, deer were euthanatized, examined, and tissues were collected 84 to 87 days post inoculation. Possible routes of disease transmission were evaluated by culture of nasal, oral, tonsilar, and rectal swabs at various times during the study. Gross and microscopic lesions consistent with tuberculosis were most commonly seen in medial retropharyngeal lymph nodes and lung in both dosage groups. Other tissues containing tuberculous lesions included tonsil, trachea, liver, and kidney as well as lateral retropharyngeal, mandibular, parotid, tracheobronchial, mediastinal, hepatic, mesenteric, superficial cervical, and iliac lymph nodes. Mycobacterium bovis was isolated from tonsilar swabs from 8 of 9 deer from both dosage groups at least once 14 to 87 days after inoculation. Mycobacterium bovis was isolated from oral swabs 63 and 80 days after inoculation from one of three deer in the low dose group and none of four deer in the high dose group. Similarly, M. bovis was isolated from nasal swabs 80 and 85 days after inoculation in one of three deer from the low dose group and 63 and 80 days after inoculation from two of four deer in the high dose group. Intratonsilar inoculation with M. bovis results in lesions similar to those seen in naturally infected white-tailed deer; therefore, it represents a suitable model of natural infection. These results also indicate that M. bovis persists in tonsilar crypts for prolonged periods and can be shed in saliva and nasal secretions. These infected fluids represent a likely route of disease transmission to other animals or humans. PMID- 10479079 TI - Two Theileria cervi SSU RRNA gene sequence types found in isolates from white tailed deer and elk in North America. AB - Two Theileria cervi SSU rRNA gene sequence Types, F and G, from white-tailed deer (Odocoileus virginianus) and elk (Cervus elaphus canadensis) isolates in North America were confirmed. Previously, nucleotide sequencing through a single variable (V4) region showed the presence of SSU rRNA gene Types F and G in T. cervi isolates from white-tailed deer and an elk. In this study, both sequence types were found in four T. cervi isolates (two from deer and two from elk). Microheterogeneity only appeared in the Type G gene, resulting in Subtypes G1, G2 and G3. Subtype G1 was found in two elk and one white-tailed deer T. cervi isolate; Subtypes G2 and G3 were found in a white-tailed deer T. cervi isolate. The Type F SSU rRNA genes were identical in nucleotide sequence in both elk and white-tailed deer T. cervi isolates. The high degree of conservation in the Type F variable regions may be exploited to design specific oligonucleotide primers for parasite detection by the polymerase chain reaction in cervine or tick hosts. PMID- 10479080 TI - An experimental study on the effects of polymorphiasis in common eider ducklings. AB - Eight common eider (Somateria mollissima) ducklings were experimentally infected from 1 June through 13 June, 1995 with acanthocephalans (Polymorphus minutus) by allowing the birds to feed on Gammarus spp. (Gammarus oceanicus, G. salinus, G. zaddachi, and G. lacustris) containing acanthocephalan cystacanths. Uninfected Gammarus spp. were fed to a control group of seven ducklings. No mortality of ducklings occurred during the experiment. However, the infected ducklings gained weight more slowly than the control birds. After the 2 wk study period, the mean serum concentrations of total protein, albumin, beta-globulin, gamma-globulin, fructosamine and creatine kinase were lower in the infected group than in the controls. The mean (+/-SE) number of acanthocephalans in the intestine of the infected ducklings was 21 (+/-4). The parasites were attached to the mucosa of the posterior small intestine of the infected ducklings with a mixed inflammatory reaction consisting of heterophils and mononuclear lymphocytes surrounding the attachment sites. PMID- 10479081 TI - Implications of raccoon latrines in the epizootiology of baylisascariasis. AB - Raccoons (Procyon lotor) frequently establish defecation sites, termed latrines, on large logs, stumps, rocks, and other horizontally oriented structures/surfaces. These latrines are important foci of infective eggs of Baylisascaris procyonis, a nematode parasite of raccoons which is pathogenic to numerous species of mammals and birds. To examine the role of raccoon latrines in this animal-parasite interaction, we documented animal visitations to raccoon latrines in two large forested tracts and two woodlots in Indiana (USA) during 1994 and 1995. Species richness of vertebrate visitors did not differ between sites or years, but species composition differed by site and year. Fourteen mammal and 15 bird species were documented visiting raccoon latrines. Small granivorous mammals, including white-footed mice (Peromyscus leucopus), eastern chipmunks (Tamias striatus), and tree squirrels (Sciurus carolinensis, S. niger, Tamiasciurus hudsonicus) were the most common visitors to latrine sites. White footed mice, chipmunks, white-breasted nuthatches (Sitta carolinensis), and opossums (Didelphis virginiana) were photographed foraging on undigested seeds in raccoon feces. Active foraging at latrines also was shown experimentally; seeds embedded in raccoon feces were removed at a greater rate at latrine sites than at nonlatrines. We conclude that raccoon latrines are visited routinely by a variety of vertebrates, especially small granivorous rodents and birds which forage for seeds in raccoon feces, and that raccoon latrines are probable sites of transmission of B. procyonis to susceptible mammals and birds. PMID- 10479082 TI - Aspergillosis and other causes of mortality in the stitchbird in New Zealand. AB - Necropsy findings from natural deaths in free living and captive stitchbirds (Notiomystis cincta) were examined over a 3 yr period (November 1991-94) to establish whether disease was an important factor in translocation failures and captive breeding programs undertaken by the New Zealand Department of Conservation. Fresh and fixed material from seven free-living birds and 11 captive birds were examined and were compared with those of a retrospective study of archival material from captive and wild birds collected over a 13 yr period (1979-91). The causes of death in both the present and retrospective study showed a similar pattern with aspergillosis and aspiration pneumonia being the most significant cause of mortality in captive birds. Aspergillosis was diagnosed as the cause of death in 11 of 31 stitchbirds from Mt Bruce; eight of these deaths occurred in the winter months (June-August). The other causes of death in captive birds included trauma, coccidiosis, and sporadic bacterial infections. Hemosiderosis and airsacculitis were common histological findings in most of the wild and captive stitchbirds examined. PMID- 10479083 TI - Mass stranding of wedge-tailed shearwater chicks in Hawaii. AB - Unusual numbers of wedge-tailed shearwater (Puffinus pacificus) chicks stranded on Oahu (Hawaii, USA) in 1994. Compared to healthy wedge-tailed shearwater (WTSW) chicks, stranded chicks were underweight, dehydrated, leukopenic, lymphopenic, eosinopenic, and heterophilic; some birds were toxemic and septic. Stranded chicks also were hypoglycemic and had elevated aspartate amino transferase levels. Most chicks apparently died from emaciation, dehydration, or bacteremia. Because many birds with bacteremia also had severe necrosis of the gastrointestinal (GI) mucosa associated with bacteria, we suspect the GI tract to be the source of disseminated bacterial infection. The identity of the bacteria was not confirmed. The daily number of chicks stranded was significantly related to average wind speeds, and the mortality coincided with the fledging period for WTSW. Strong southeasterly winds were a distinguishing meteorologic factor in 1994 and contributed to the distribution of stranded chicks on Oahu. More objective data on WTSW demographics would enhance future efforts to determine predisposing causes of WTSW wrecks and their effects on seabird colonies. PMID- 10479084 TI - Health evaluation of a pronghorn antelope population in Oregon. AB - During 1996 and 1997, the U.S. Fish and Wildlife Service conducted a study to determine the cause(s) of population decline and low survival of pronghorn antelope (Antilocapra americana) fawns on Hart Mountain National Antelope Refuge (HMNAR) located in southeastern Oregon (USA). As part of that study, blood, fecal, and tissue samples from 104 neonatal fawns, 40 adult does, and nine adult male pronghorns were collected to conduct a health evaluation of the population. Physiological parameters related to nutrition and/or disease were studied. No abnormalities were found in the complete blood cell counts of adults (n = 40) or fawns (n = 44 to 67). Serum total protein and blood urea nitrogen (BUN) levels were lower compared to other pronghorn populations. Does had mean BUN values significantly lower (P < 0.001) in December 1996 than March 1997. Serum copper (Cu) levels in does (range 0.39 to 0.74 ppm) were considered marginal when compared to domestic animals and other wild ungulates. Fawns had low (0.28 ppm) Cu levels at birth and reached the does' marginal values in about 3 days. Whole blood, serum and liver selenium (Se) levels were considered marginal to low in most segments of the pronghorn population. However, serum levels of vitamin E (range 1.98 to 3.27 microg/ml), as determined from the does captured in March, were apparently sufficient to offset any signs of Se deficiency. No clinical signs of Cu or Se deficiency were observed. Fifty-five of 87 dead fawns were necropsied. Trauma, due to predation by coyotes (Canis latrans), accounted for 62% of the mortality during mid-May to mid-July of each year. Other causes included predation by golden eagles (Aquila chrysaetos) (4%), dystocia (2%), septicemic pasteurellosis (4%), starvation (5%), and unknown (23%). Adult females were tested for serum neutralizing antibodies to Brucella spp. (n = 20, negative), Leptospira interrogans (n = 20, negative), bluetongue virus (n = 20, 35% positive), epizootic hemorrhagic disease virus (n = 20, 30% positive), respiratory syncytial virus (n = 18, negative), parainfluenza virus type 3 (n = 18, 67% positive), infectious bovine rhinotracheitis (n = 18, negative), and bovine viral diarrhea (n = 18, negative). Considering the parameters examined, we found no apparent predisposing factors to mortality including those killed by coyotes, but some nutritional parameters suggest that pronghorns on HMNAR exist on a diet low in protein and Se and marginal in Cu. The effect these factors have on the population is not known. PMID- 10479085 TI - Synovial joint disease in wild cetaceans. AB - During necropsy of cetaceans stranded or accidentally net-captured along the western coast of the Gulf of Mexico from 1991 to 1996, we found 13 of 59 (22%) animals had abnormalities of the atlanto-occipital and/or humeroscapular joints, the synovial joints. A few cases demonstrated mild roughening of the articular cartilage, while the majority exhibited complete erosion with thickened synovium and bony proliferation. The lesions resulted in ankylosis of both joints in one animal. In humans and terrestrial mammals, synovial joint diseases are known to be debilitating. Cetaceans depend on neck and flipper movement for locomotion, feeding, avoiding danger, and reaching the water's surface for breathing. Therefore, synovial joint disease may be significant mortality factor in these marine animals. PMID- 10479086 TI - Atrial septal defects in Florida panthers. AB - Ostium secundum atrial septal defects (ASDs) were observed in six (3 M, 3 F) of 33 (20 M, 13 F) (18%) Florida panthers (Puma concolor coryi) necropsied by veterinary pathologists between 1985 and 1998. A seventh ASD was found in a female panther necropsied in the field and is included in the pathological description but not the prevalence of ASDs in Florida panthers. One panther (FP205) with severe ASD also had tricuspid valve dysplasia (TVD). Atrial septal defects and/or TVD are believed to have caused or contributed to the deaths of three (9%) Florida panthers in this study. Mean diameter +/- SD of ASDs was 9.0 +/- 4.7 mm (range 3 to 15 mm). Gross pathological changes attributed to ASDs/TVD in severely affected panthers (ASD > or = 10 mm) (n = 4) included mild right ventricular dilatation (n = 3) and hypertrophy (n = 2), mild to severe right atrial dilatation (n = 2), and acute pulmonary edema (n = 3). Panthers with mild ASDs (ASD < or = 5 mm) (n = 3) had no other detectable gross pathological changes associated with the ASDs. Histological examination of lungs of three panthers with severe ASDs revealed mild to moderate dilatation with fibrosis and smooth muscle atrophy of the tunica media of medium to large caliber arteries (n = 2), interstitial and/or pleural fibrosis (n = 2), perivascular fibrosis (n = 1), and acute to chronic edema (n = 3). Twenty-six necropsied panthers were examined one or more times while living; medical records were retrospectively evaluated. Antemortem radiographic, electrocardiographic, and echocardiographic examinations were performed on two panthers with severe ASDs (FP20 and FP205). Thoracic radiographic abnormalities in both included right heart enlargement, and in FP205 (severe ASD and TVD), mild pulmonary overperfusion. Electrocardiographic examination of FP205 revealed a right ventricular hypertrophy pattern, while FP205 had a normal electrocardiogram. Echocardiographic examination of FP20 revealed marked right atrial dilatation; a bubble contrast study indicated regurgitation across the tricuspid valve. Echocardiographic abnormalities in FP20 included right atrial and ventricular lilatation, atrial septal drop-out, and severe tricuspid regurgitation; non-selective angiography revealed significant left to right shunting across the ASD. All panthers with severe ASDs ausculted (n = 3) had systolic right or left-sided grade I-V/VI murmurs loudest at the heart base. All male panthers with ASDs (n = 3) (100%) and 9 of 17 (53%) male panthers without ASDs in this study were cryptorchid. PMID- 10479087 TI - Hematological and serum chemistry profiles of free-ranging southern two-toed sloths in French Guiana. AB - Free-ranging southern two-toed sloths (Choloepus didactylus) were translocated during the flooding of a forest at a hydroelectric dam site in French Guiana. Over an 11 mo period blood samples were collected from 90 sloths (38 males, 52 females) in order to determine hematological and serum chemistry reference values. Mean values and range of values were calculated for 13 hematological and 21 serum chemistry parameters. Variations associated with sex, age and reproductive status were identified. Males had a significantly lower red blood cell count than females. Immature animals had more monocytes while adults had more neutrophils and higher mean corpuscular hemoglobin concentration. Aspartate aminotransferase and triglyceride values were higher in young than in adult sloths but uric acid was lower. Lactating females showed lower red blood cells count and iron levels than non-lactating females. These profiles will help to provide reliable baseline data for medical evaluation of sloths. PMID- 10479088 TI - Effects of malathion on disease susceptibility in Woodhouse's toads. AB - Adult male Woodhouse's toads (Bufo woodhousi) developed clinical disease, hepatomegaly, and died at a higher rate when externally exposed once to either a high or low sublethal dose (0.011 or 0.0011 mg malathion/g toad) of field grade malathion and challenged with a sublethal dose of Aeromonas hydrophila injected intraperintoneally (1.1 x 10(4) bacteria/g toad) when compared to toads not exposed to malathion but challenged with A. hydrophila (P < 0.007). Toads exposed to malathion (high or low dose) and challenged with A. hyydrophila had clinical disease, hepatomegaly, and died at a higher rate [9 (90%) of 10] than toads exposed to malathion alone (P < 0.002). Toads exposed to the high and low doses of malathion had a 22% and 17% decrease in brain cholinesterase levels, respectively, when they were compared to nonmalathion exposed toads (P < 0.025, P < 0.006). It appears that field grade malathion applied externally to adult Woodhouse's toads may cause increased disease susceptibility when challenged with a potentially pathogenic bacteria. PMID- 10479089 TI - Exertional myopathy in translocated river otters from New York. AB - Lesions consistent with exertional myopathy (EM) were documented postmortem in four North American river otters (Lutra canadensis) during translocation for a population restoration project. Clinical signs in these otters included depression, anorexia and shock. Gross lesions in one otter included locally extensive linear, pale areas within the subscapularis, rectus abdominis, quadriceps, and dorsal laryngeal muscles. Microscopic lesions were characterized by acute to subacute myofiber necrosis of varying severity, and occurred in a variety of skeletal muscles as well as cardiac muscle in one otter. Based on these observations, we conducted a retrospective review of records of otters which experienced similar capture, transfer, and holding protocols between 1995 and 1997, but with a successful outcome (n = 69). Significant elevations in serum aspartate aminotransferase (AST) and creatine kinase (CK) were observed in 19 (28%) of the otters, but may have been higher due to delayed sample collection from some otters. However, none of the otters with elevated enzymes exhibited clinical signs suggestive of EM. These findings indicate that river otters may develop EM when translocated, but many cases may be mild or clinically inapparent. PMID- 10479090 TI - Cardiopulmonary response of anesthetized polar bears to suspension by net and sling. AB - Since 1995, at least three polar bears (Ursus maritimus) have died in the area of Churchill (Manitoba, Canada) as a direct result of being suspended in a net during helicopter-assisted translocations. To assess and improve methods of suspending anesthetized polar bears, we conducted a study during November 1997 to determine the cardiopulmonary responses of eight captive polar bears to suspension by net and by sling. Each bear was anesthetized on two occasions in which the sequence of activities followed and the type of data collected was identical, with only the method of suspension differing. Control data obtained from 11 captive polar bears during 1995-96 was included in the statistical analyses of cardiopulmonary data to help clearly differentiate the cardiopulmonary effects of suspension from those of drug metabolism. Suspending polar bears above the ground by net caused acute hypertension (e.g., 17 to 49% increase in mean arterial pressure), possibly as a result of increased venous return due to body compression. Increased arousal (e.g., head, tongue, and limb movement) also occurred consistently during net-suspension and suggested a stress response. Surprisingly, most suspended bears showed little change in blood gas values, but at least one bear became hypoxemic (i.e., PaO2 < 60 mm Hg) with each method of suspension. Because of the potential health risks of hypertension and hypoxemia, we recommend modifying the method by which polar bears are suspended with the goal of reducing body compression. PMID- 10479091 TI - Bat rabies in South Carolina, 1970-90. AB - This retrospective study examines the geographic and temporal distribution of bat species submitted for rabies testing in South Carolina (USA) from 1970 to 1990. Histories of human or animal exposures to rabid and nonrabid bats submitted during this time period were compared. Two hundred and thirty-one bats were found to be rabid from the 2,657 bats submitted over this 20 yr period. The two species most frequently submitted for testing were Lasiurus borealis with 785 specimens (30% of the total) and Nycticeius humeralis with 607 specimens (22% of the total). Lasiurus borealis also had the highest prevalence of rabies (18%) while N. humeralis had the lowest prevalence (3%). Fifty-one percent (1,259) of the bats received for testing were submitted from June through August. The majority (54%) of rabid bats were received from August through October. PMID- 10479093 TI - Rhizopusmycosis in a harbor porpoise from the Baltic Sea. AB - A case of systemic mycosis due to a Rhizopus sp. infection is described in a dead stranded, 10-yr-old, male harbor porpoise (Phocoena phocoena) found on the beach of Neustadt, Schleswig-Holstein on the Baltic Sea (Germany). At necropsy, granulomatous mycotic lesions in brain, lung, kidneys, testis, and draining lymph nodes were found. In addition, a focal ulcerative gastritis of the first stomach, due to a nematode infection, was present and is suspected to be the portal of entry for the fungus. PMID- 10479092 TI - Hepatic carcinoma with spleen metastasis in a California sea lion from the Gulf of California. AB - A primary hepatic carcinoma with a neuroendocrine pattern was detected in an adult female California sea lion (Zalophus californianus) found dead on Granito Island in the Gulf of California (Mexico) in January 1996. At necropsy, several light yellow nodules of different sizes were observed on the entire surface of the liver and spleen. Microscopic examination of these nodules using routine haematoxylin-eosin stain, revealed cubic, polyhedral and pleomorphic cells with three to four bizarre mitotic figures per field (40X). An immunohistochemistry test revealed a positive reaction of indirect immunoperoxide to cytokeratin (CK2). This is the first known case of a primary hepatic carcinoma in free ranging California sea lions from Mexican waters. PMID- 10479094 TI - Rabies in skunks from Mexico. AB - An enzootic focus of rabies in skunks in Mexico is described. Fifty three wild animals including two badgers (Taxidea taxus), 32 bats (various species), one bobcat (Lynx rufus), two coatis (Nasua narica) three foxes (Urocyon cineroargenteus), one raccoon (Procyon lotor) and 12 skunks (see below) were tested for rabies by direct immunofluorescence assay from 1991 to 1997 in the central part of San Luis Potosi State, Mexico. Rabies occurrence was 21% of all tested mammals, with 19% in skunks and only 2% in other wild species (one bobcat). Skunks represented 23% of all mammals tested and had a rabies prevalence of 83%. Only 10 individuals were identified: three hog-nosed skunks (Conepatus leuconotus) and seven spotted skunks (Spilogale putorius). All were involved in human attacks; the spotted skunk attacks were inside bedrooms while people were sleeping, and the hog-nosed skunk attacks occurred outdoors. Skunk cases of rabies represented 40% of all rabies cases in 1997, and 100% of cases registered for wild animals in San Luis Potosi state. This situation constitutes an important public health problem and requires further epidemiological research to make the human population aware of the problem and to establish measures to limit further human attacks by rabid skunks. PMID- 10479095 TI - Serosurvey for selected virus infections of wild carnivores in Taiwan and Vietnam. AB - Serum samples from two leopard cats (Felis bengalensis) and four Formosan gem faced civets (Paguma larvata taivana) in Taiwan, September 1995, and nine leopard cats in Vietnam, August and December 1997, were examined for the prevalence of antibodies against feline parvovirus, feline herpesvirus type 1, feline calicivirus and feline immunodeficiency virus. All civets and nine of 11 leopard cats were shown to have antibodies against feline parvovirus (FPV), and FPV's were isolated from mononuclear cells in the peripheral blood of the six leopard cats. PMID- 10479096 TI - Avian pox in sanderlings from Florida. AB - Asian pox was diagnosed in three sanderlings (Calidris alba) on Sanibel Island, Florida (USA) in February 1997. All three cases had large tumor-like lesions which contributed significantly to their mortality. Poxvirus infection was confirmed by cytology, histopathology, and electron microscopy. This is the first report of poxvirus infection in sanderlings. PMID- 10479098 TI - Use of western immunoblot analysis for testing moose serum for Brucella suis biovar 4 specific antibodies. AB - To determine if 12 moose (Alces alces) from northern Alaska with agglutinating antibodies specific for Brucella spp. had been exposed to either B. suis biovar 4 or B. abortus biovar 1, western immnnoblot serologic analysis was performed. Differential serologic responses to strain specific A and M antigenic variances of the lipopolysaccharide O-polysaccharide sugar allowed strain identification. Prior to examination, test sera were absorbed with killed whole cells from either B. abortus biovar 1, containing predominately A antigen (A+ M-); B. melitensis biovar 1, containing essentially M antigen (A- M+); or B. suis biovar 4, containing both antigenic tyes (A+ M+). The resulting sera were then examined by western immunoblot for recognition of either B. abortus biovar 1, B. melitensis biovar 1, or B. suis biovar 4 cell lysates. The results of this study indicate that these moose were exposed to B. suis biovar 4, a known pathogen of caribou (Rangifer tarandus) from arctic Alaska. PMID- 10479097 TI - Mycoplasmosis associated perosis type skeletal deformity in a saker falcon nestling in Hungary. AB - A wild, 3-wk-old saker falcon (Falco cherrug) nestling showing uncoordinated movements and a perosis type tarsometatarsus deformity was found abandoned; it was euthanized a week later on 29 May 1997 after an unsuccessful attempt to rehabilitate it. Gross pathological findings included congestion of parenchymal organs and a lateral bowing of the left tarsometatarsal bone. Histopathology revealed initial interstitial hepatitis, focal catarrhal pneumonia, and dyschondroplasia in the epiphysis of the left tarsometatarsus. Mycoplasmas were isolated from the lungs, trachea, bone marrow and brain. A polymerase chain reaction (PCR) assay was performed for the detection of the mycoplasmal 16S rRNA gene. The resulting 262 base pair PCR product was sequenced and compared to the available mycoplasmal sequences but no identical corresponding sequences were found. However, 98% similarity was found to the Mycoplasma buteonis 16S rRNA and the isolate also was positive by immunoblotting against reference sera to the same species. PMID- 10479099 TI - Ehrlichia chaffeensis in archived tissues of a white-tailed deer. AB - White-tailed deer (Odocoileus virginianus) play an integral role in the natural history of Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis (HME). Paraffinized tissues from a white-tailed deer submitted as a diagnostic case to the Southeastern Cooperative Wildlife Disease Study (Athens, Georgia, USA) in October of 198.5 and originally described as infected with an unidentified rickettsial organisim were re-examined by specific nested polymerase chain reaction (PCR) for evidence of infection with Ehrlichia spp. Ehrlichia chaffeensis was identified from the bone marrow and inguinal lymph node of this deer based on amplification of a characteristic sequence-confirmed 16S rDNA fragment from these tissues. Parallel PCR tests on the same samples were negative for 16S rDNA fragments of the agent of human granulocytic ehrlichiosis (HGE) and for an Ehrlichia-like organism widely distributed in white-tailed deer populations. This report describes detection of E. chaffeensis in archived tissue from a deer collected before the index case of human monocytic ehrlichiosis was established. PMID- 10479100 TI - Plague in free-ranging mammals in western North Dakota. AB - From July through October of 1996, 48 blood samples were collected from coyotes (Canis latrans), badgers (Taxidea taxus), and raccoons (Procyon lotor) in western North Dakota (USA) for the purposes of determining antibody titers to the plague bacterium, Yersniia pestis. The passive hemagglutination paper-strip blood sampling technique was utilized with hemagglutination inhibition controls. Two positive samples were obtained from McKenzie county, one from a coyote with a titer of 1:64 and one from a badger with a titer of 1:256. Considering coyote and badger population dynamics, this study documents plague in western North Dakota. PMID- 10479101 TI - Leucocytozoonosis and trypanosomiasis in redstarts in Finland. AB - Leucocytozoon spp. and Trypanosoma spp. blood parasites in the redstart (Phoenicurus phoenicurus) were studied during spring migration 1994 in southern Finland (53 individuals) and the breeding season 1992-1994 in northern Finland (69). Parasite prevalence was higher during the breeding season (48%) than during the migration period (13%), with no age or sex differences in the breeding site birds. In both periods, redstarts were infected by the same blood parasites Leucocytozoon shaartusicum (46% prevalence at the breeding site and 71% during the migration period) and Trypanosoma avium, complex (58% and 43%, respectively). One individual at the breeding site had contracted L. dubreuili and one at the stop-over site had T. everetti. Our results may support the assumption that tissue-hidden parasites relapse during the breeding season when birds may have diminished immune response related to egg production and brood rearing. Another explanation could be that the high abundance of ornithophilic vectors enhance parasite transmission during breeding season in northern Finland. PMID- 10479102 TI - Leucocytozoonosis in nestling bald eagles in Michigan and Minnesota. AB - Thirteen of 21 nestling bald eagles (Haliaeetus leucocephalus) examined for blood parasites in Michigan and Minnesota (USA) during June and July 1997 had patent infections of Leucocytozoon toddi. No other parasites were seen. The degree of parasitemia was light and varied from 1 to 2 on the Ashford Scale. Several of the infected nestlings appeared to have elevated levels of heterophils in their peripheral circulating blood. One of the infected nestlings also showed signs of severe anemia. We believe this is the first report of L. toddi in the bald eagle. PMID- 10479103 TI - Hematologic effects of cytauxzoonosis in Florida panthers and Texas cougars in Florida. AB - Cytauxzoon felis is a long-recognized hemoparasite of free-ranging Florida panthers (Puma concolor coryi), but its prevalence and effect on the population has not been assessed. Red blood cell indices and white blood cell counts were compared between infected and noninfected Florida panthers and Texas cougars (Puma concolor stanleyana) from 1983-1997 in Florida (USA). The prevalence of cytauxzoonosis for both populations was 39% (11/28) for Texas cougars, 35% for Florida panthers (22/63) and 36% overall. Thirteen hematologic parameters were compared between C. felis positive and negative panthers and cougars. Florida panthers had significantly lower mean cell hemoglobin count (MCHC) and higher white blood cell (WBC), neutrophil, monocyte and eosinophil counts (P < or = 0.05) than Texas cougars. Infected Florida panthers had significantly lower mean cell hemoglobin (MCH) and monocyte counts and higher neutrophil and eosinophil counts than infected Texas cougars. Although statistically significant differences were measured for hematologic parameters in C. felis positive panthers and cougars, biologically significant differences were not likely because values were generally within expected reference ranges for healthy animals. Cytauxzoonosis does not appear to have a negative effect on the hematologic parameters of chronically infected panthers and cougars. Potential transient changes during initial infection were not evaluated. PMID- 10479104 TI - Parasites in grizzly bears from the central Canadian Arctic. AB - Standardized flotation techniques were used to survey 56 grizzly bear (Ursus arctos) fecal samples for parasites. The samples were collected during the spring and autumn of 1995 and 1996 in the central Arctic of the Northwest Territories (Canada). Parasites of the genera Nematodirus, gastrointestinal coccidia, and an unidentified first stage protostrongylid larva are reported for the first time from grizzly bear feces in North America. Parasites of the genera Diphyllobothrium and Baylisascaris also were collected. Prevalence of gastrointestinal parasites were significantly different between the spring and autumn seasons (31% and 58% respectively). Thus, we provide evidence supporting the theory that bears void gastrointestinal parasites before hibernation. PMID- 10479105 TI - Search for the trematode Prouterina wescotti in black bears in Oregon. AB - Forty-six heads of free-ranging black bears (Ursus americanus) collected in May and June of 1995 and 1996 from Oregon (USA) were examined for the trematode Prouterina wescotti and other parasites. Only Sarcocystis sp. was detected in two adult male bears. Prouterina wescotti has been found only once previously in one black bear from Idaho (USA); its normal definitive host remains unknown. PMID- 10479106 TI - Health status of a recently discovered population of feral swine in Kansas. AB - Twenty feral hogs (Sus scrofa) from a newly discovered population on Fort Riley Army Base (Kansas, USA) were shot and examined from November 1993 through February 1994 to assess the health of the population. The hogs were generally healthy, although serologic evidence indicated that some individuals had been exposed to parvovirus, enterovirus, and swine influenza. We found no indications of brucellosis, pseudorabies, or porcine respiratory and reproductive syndrome. Lung worms (Metastrongylus spp.), round worms (Ascaris suum), and whipworms (Trichuris suis) were found in nine, four and two of the hogs, respectively. Seven hogs had infestations of lice (Haematopinus suis). Fence-line contacts were documented between four wild boars and domestic sows, and in three cases wild boars entered pens containing domestic sows. We recommend that hogs be examined periodically from this and other wild populations to monitor health status since new animals may enter populations through deliberate translocation, escape from shooting preserves or domestic swine producers, or dispersal from other feral populations. PMID- 10479107 TI - Telehealth and state licensure. PMID- 10479108 TI - Managing alcohol-related problems in the primary care setting. AB - Although alcohol-related problems are a significant source of morbidity and mortality in the United States, denial and resistance to treatment by patients and their families make detection challenging. Primary care providers may find it difficult to confront the patient who drinks excessively; however, early intervention can lower morbidity and prevent progressive damage to family and social relationships. Routine alcohol screening of all patients may assist the primary care provider in effectively identifying and treating alcohol abuse. Although abstinence is the primary goal for patients with alcohol-related problems, decreasing intake is also beneficial and may be accomplished with brief primary care interventions. The management of outpatient alcohol detoxification is increasingly provided by primary care providers in managed care environments, but requires careful assessment of the patient's support system, close provider supervision, and appropriate pharmacologic support. PMID- 10479109 TI - Recognizing the various presentations of appendicitis. AB - Appendicitis is one of the most common causes of acute abdominal pain in the industrialized world. Appendicitis must be considered in the differential diagnosis of any patient presenting with abdominal pain. Workup may include blood tests, abdominal radiographs, abdominal ultrasound, and focused appendix computed tomography. Unfortunately, none of these provides definitive results. Although several signs and symptoms are associated with appendicitis, their inconsistent presentation, especially among the young and the elderly, can lead to an erroneous diagnosis. The classic sequence of symptoms includes the onset of vague epigastric or periumbilical pain; associated nausea, anorexia, or unsustained vomiting; and pain migrating to the right lower quadrant. In uncomplicated cases, the treatment of appendicitis is appendectomy. However, less definitive presentations merit further diagnostic testing and close follow-up. PMID- 10479110 TI - Identifying diabetic patients at high risk for amputation. AB - Lower-extremity amputation is a much feared complication of diabetes mellitus; however, 40% to 50% of these amputations are preventable. Peripheral neuropathy has been implicated as a cause in some 82% of diabetic amputations. Patient education on foot care is often not enough to prevent foot ulceration and potential amputation. Health care providers can lower the incidence of lower extremity amputation by using a Semmes-Weinstein monofilament to identify protective sensation loss and quickly taking measures to prevent ulceration. Diabetes patients who have a history of foot deformity, ulceration, or amputation of any part of the foot should be referred for special shoes or orthotics. This article reviews the methods for identifying those patients with diabetes that are at high risk for amputation and preventive interventions. PMID- 10479111 TI - Landmark Maine bill signed into law. PMID- 10479112 TI - Cervical dysplasia in women with HIV. AB - Prevention of cervical cancer in HIV-positive women is possible through regular cervical cytological screening for early detection and a variety of effective treatment options for dysplasia. Women with HIV infection continue to have an increased rate of CIN, and cervical dysplasia appears to recur more frequently and progress more rapidly in severely immunocompromised women with high-risk type HPV infection. The Pap test is an effective screening tool, but high rates of recurrence in immunocompromised women reinforce the need for more frequent screening in HIV-positive women with a history of abnormal smears and CD4 counts less than 200. Health care providers should be aware of the latest recommendations for cervical dysplasia evaluation and should apply these in their care of HIV-infected women. Early detection, prompt treatment, and meticulous follow-up are essential in the prevention of cervical dysplasia, as well as the morbidity and mortality associated with cervical cancer in women with HIV. PMID- 10479113 TI - Nonscientifically validated herbal treatments for vaginitis. PMID- 10479114 TI - Longitudinal survey of acute care nurse practitioners. PMID- 10479115 TI - Low incidence of rimantadine resistance in field isolates of influenza A viruses. AB - The spread of drug-resistant influenza viruses type A to close contacts in families, schools, and nursing homes has been well documented. To investigate whether drug-resistant influenza viruses circulate in the general population, 2017 isolates collected in 43 countries and territories during a 4-year period were tested for drug susceptibility in a bioassay. Drug resistance was confirmed by detection of specific mutations on the M2 gene that have been shown to confer resistance to amantadine or rimantadine. Sixteen viruses (0.8%) were found to be drug-resistant. Only 2 of these resistant viruses were isolated from individuals who received amantadine or rimantadine treatment at the time the specimens were collected. For 12 individuals use of amantadine or rimantadine could be excluded, and from the remaining 2 patients information about medication was unavailable. These results indicate that the circulation of drug-resistant influenza viruses is a rare event, but surveillance for drug resistance should be continued. PMID- 10479116 TI - Intranasal interleukin-12 is a powerful adjuvant for protective mucosal immunity. AB - The use of interleukin (IL)-12 as a new vaccine adjuvant for stimulating protective antiviral mucosal immunity has been examined. Mice were immunized intranasally (in) with an influenza vaccine consisting of soluble hemagglutinin (H1) and neuraminidase (N1) plus IL-12. This treatment resulted in elevated levels of lung and splenic interferon-gamma and IL-10 mRNA. Total and IgG2a anti H1N1 antibody levels in serum were significantly elevated, as were total, IgG1, IgG2a, and secretory IgA antibody levels in bronchoalveolar lavage (BAL) fluids compared with animals receiving vaccine alone. Mice immunized in with vaccine and IL-12 also exhibited decreased weight loss and dramatically enhanced survival after lethal challenge with infectious influenza virus. Protection was dependent upon the presence of B cells and could be transferred to naive mice by inoculation of either serum or BAL fluid from IL-12-treated mice. These findings show for the first time that soluble IL-12 delivered in serves as a powerful respiratory adjuvant for protective antiviral immunity. PMID- 10479117 TI - Measles virus infection in rhesus macaques: altered immune responses and comparison of the virulence of six different virus strains. AB - Measles remains a major cause of childhood mortality, with questions about virus virulence and pathogenesis still requiring answers. Rhesus macaques were infected with 5 different culture-adapted strains of measles virus, including 2 from patients with progressive vaccine-induced disease, and a sixth nonculture-adapted strain, Bilthoven. All caused infection detectable by reverse transcriptase polymerase chain reaction and induction of antibody. Chicago-1 and Bilthoven induced viremias detectable by leukocyte cocultivation. Bilthoven induced Koplik's spots, conjunctivitis, and rash. Lymphopenia and depressed interleukin (IL)-2 production were followed by monocytosis and eosinophilia. All monkeys, including 41 involved in a primate facility outbreak, showed suppressed responses to phytohemagglutinin. As the rash resolved production of IL-2, IL-1beta, tumor necrosis factor-alpha, IL-6, and IL-5 mRNA increased. Monkeys are useful for studies of measles immunopathogenesis, but virus strains must be carefully chosen. Increased virulence of vaccine strains isolated from immunocompromised infants with fatal infections was not evident. PMID- 10479118 TI - Partial nucleotide sequencing and molecular evolution of epidemic causing Dengue 2 strains. AB - To study the genetic variability and to detect evolutionary changes and movement of dengue 2 (DEN-2) strains, nucleotide sequencing of the envelope protein gene and the nonstructural protein 1 gene junction was performed for 9 isolates from the 1996 Delhi epidemic and 1 isolate from the 1967 Delhi epidemic. The epidemic strains had a divergence of 10%-11% from the 1967 strains, but were quite similar to DEN-2 isolates from Seychelles, Somalia, and Torres Strait. In addition, the sequence data were compared to the prototype DEN-2 strain, New Guinea C, and other published DEN-2 sequences from different parts of the world. The phylogenetic analysis by the Molecular Evolutionary Genetics Analysis program suggests that the 1996 Delhi isolates of DEN-2 were genotype IV. The 1967 isolate was similar to a 1957 isolate of DEN-2, P9-122, from India, and was classified as genotype V. This study indicates that earlier DEN-2 strains of genotype V have been replaced by genotype IV. PMID- 10479119 TI - Experimental infection of the cane mouse Zygodontomys brevicauda (family Muridae) with guanarito virus (Arenaviridae), the etiologic agent of Venezuelan hemorrhagic fever. AB - Chronic infections in specific rodents appear to be crucial to the long-term persistence of arenaviruses in nature. The cane mouse, Zygodontomys brevicauda, is a natural host of Guanarito virus (family Arenaviridae), the etiologic agent of Venezuelan hemorrhagic fever. The purpose of this study was to elucidate the natural history of Guanarito virus infection in Z. brevicauda. Thirty-nine laboratory-reared cane mice each were inoculated subcutaneously with 3.0 log10 plaque-forming units of the Guanarito virus prototype strain INH-95551. No lethality was associated with infection in any animal, regardless of age at inoculation. The 13 newborn, 14 weanling, and 8 of the 12 adult animals developed chronic viremic infections characterized by persistent shedding of infectious virus in oropharyngeal secretions and urine. These findings indicate that Guanarito virus infection in Z. brevicauda can be chronic and thus support the concept that this rodent species is the natural reservoir of Guanarito virus. PMID- 10479120 TI - A subunit cytomegalovirus vaccine based on recombinant envelope glycoprotein B and a new adjuvant. AB - A phase I randomized, double-blind, placebo-controlled trial was done with a cytomegalovirus (CMV) vaccine based on the envelope glycoprotein, gB, combined with a novel adjuvant, MF59. Participants received CMV gB vaccine with MF59 or CMV gB with alum or placebo at 0, 1, and 6 months. A fourth vaccine was given at 12 months to a subgroup. Levels of neutralizing antibody and antibody to gB 2 weeks after the third dose of vaccine exceeded those in seropositive control subjects. the formulation with MF59 was more immunogenic than that with alum. The optimal dose of gB appeared to be between 5 and 30 microg. The fourth dose produced a prompt rise in antibody level. There were no serious adverse events associated with vaccine. Local and systemic reactions were generally mild and, except for pain at the injection site, occurred with similar frequency in recipients of placebo and CMV vaccine. PMID- 10479121 TI - Hepatocytes are permissive for human cytomegalovirus infection in human liver cell culture and In vivo. AB - The cytopathic potential of human cytomegalovirus (HCMV) in human liver cells was analyzed in cell culture and in tissue sections from patients with HCMV hepatitis. Liver cell cultures, consisting of hepatocytes, bile duct epithelial cells, and stromal cells were infected by various HCMV strains. Cytopathic effects, viral gene expression, and virus production were detected. Infected cell types were identified by immunocytochemical double labeling. Hepatocytes were the predominant target cells of HCMV infection in liver tissues and in cell culture. Late-stage infected cultured hepatocytes produced infectious progeny virus, and infectious virus was propagated from liver tissue specimens. HCMV infection in cultured liver cells closely resembled in vivo infection of the liver with regard to the target cell spectrum and the permissive course of infection. It is concluded that HCMV can cause direct liver parenchyma damage by efficient cytolytic infection of hepatocytes. PMID- 10479122 TI - Significance of the anti-E2 response in self-limited and chronic hepatitis C virus infections in chimpanzees and in humans. AB - To determine whether there was a correlation between the kinetics or frequency of antibody to mammalian-derived hepatitis C virus (HCV) second envelope protein (E2) and development of chronicity or self-limitation of HCV infections, serial sera were examined for anti-E2, anti-HCV with confirmation with Matrix 2.0 (Abbott Laboratories, Abbott Park, IL), and reverse transcriptase-polymerase chain reaction (RT-PCR) from 6 cases of self-limited infection and 6 cases of chronic infection in chimpanzees, and from 5 cases of self-limited infection and 3 cases of chronic infection in patients. Anti-E2 developed earlier, more frequently, and to higher titer in chimpanzees and patients who were developing chronic infection than in those with self-limited infections. Thus anti-E2 is unlikely to play a role in self-limitation of the infection. However, long-term persistence of anti-E2 correlates with chronic infection. There was little or no correlation between the timing of development of anti-E2 and anti-HCV. PMID- 10479123 TI - Replication of hepatitis C virus in ascitic mononuclear cells with development of distinct viral quasispecies. AB - To investigate whether distinct compositions of viral quasispecies developed in the ascitic fluid of patients with late-stage chronic hepatitis C virus (HCV) infection, samples of various origins, including ascitic fluid and ascitic mononuclear cells (AMCs), were analyzed by the method of single-strand conformation polymorphism (SSCP). Subsequently, the major species were isolated, sequenced, and subjected to phylogenetic analysis. Of 42 patients analyzed, HCV RNA was detectable in the AMCs of 25 patients. SSCP analysis indicated that the compositions of viral quasispecies among samples of different origins were markedly different in this group of patients. Phylogenetic analysis revealed that the ascitic-fluid-derived clones were most closely related to the AMC-derived clones. Minus-strand HCV-RNA was detectable in 5 of them. Our data suggest that HCV can replicate in the AMCs of patients with late-stage chronic HCV, which results in the development of distinct viral quasispecies. PMID- 10479124 TI - Differential effect of interferon on hepatitis C virus 1b quasispecies in the nonstructural protein 5A gene. AB - A close correlation was reported between amino acid mutations in the nonstructural protein 5A (interferon [IFN] sensitivity-determining region [ISDR]) of hepatitis C virus (HCV)-1b and the response to IFN therapy. The dynamic change of ISDR quasispecies during IFN treatment was investigated in 22 patients. In 18 nonresponders, the number of ISDR mutations in major quasispecies decreased after therapy (P=.039). In each nonresponder, the percentage of wild-type (no mutations in the ISDR) quasispecies increased after treatment (P=.008), whereas the percentages of intermediate- (1-3 mutations) and mutant-type (>/=4 mutations) quasispecies decreased (P=.037 and P=.043, respectively). No mutant-type quasispecies were detected after therapy. Four complete responders had only quasispecies with >/=3 mutations before therapy. Thus, HCVs have fewer mutations in the ISDR after IFN therapy than those before therapy. These IFN-resistant HCVs were already present before therapy as minor quasispecies and were selected by IFN in nonresponders. PMID- 10479125 TI - Evidence for concurrent epidemics of human herpesvirus 8 and human immunodeficiency virus type 1 in US homosexual men: rates, risk factors, and relationship to Kaposi's sarcoma. AB - We examined human herpesvirus 8 (HHV-8) seroprevalence and seroincidence among 245 homosexual men from New York City (NYC) and Washington, DC (DC) who have been followed since 1982. An immunofluorescence assay measured antibodies to a latent HHV-8 nuclear antigen. Seroprevalence was 20.4% in 1982; seroincidence was approximately 15%/year during 1982-1983 but fell sharply thereafter. NYC men had a higher seroprevalence (odds ratio, 3.43; P<.001) and seroincidence (rate ratio, 2.13; P=.01) than DC men. Risk of Kaposi's sarcoma (KS) was increased in seropositive men (adjusted relative hazard, 3.58; P=.02). Among men who were seropositive for both human immunodeficiency virus type 1 and HHV-8, the 10-year cumulative risk of KS was 39%; time from coinfection to KS diagnosis ranged from 15 to 154 months (median, 63.5 months). This study shows an epidemic of HHV-8 among US homosexual men in the early 1980s that was associated with a high risk of developing KS. PMID- 10479126 TI - Longitudinal human immunodeficiency virus type 1 load in the italian seroconversion study: correlates and temporal trends of virus load. AB - A prospective study of 149 human immunodeficiency virus type 1 (HIV-1) seroconverters was conducted to describe trends and correlates of HIV-1 load after seroconversion and over time. HIV-1 load was quantified from frozen sera by reverse transcriptase-polymerase chain reaction. High early virus load was associated with lower CD4 cell counts and male sex but not with age at seroconversion or injection drug use. Early virus load predicted progression to clinical AIDS and AIDS/<200 CD4 cells/microL. Virus load exhibited a decline of 52% by 18 months after seroconversion then increased 23% annually (95% confidence interval, 13%-33%). Men and those developing AIDS during follow-up had higher virus loads over the course of disease. Persons who developed AIDS had a steeper virus load slope than those who were AIDS-free (P=.01). In long-term follow-up, virus load exhibited a gradual and sustained increase over time. Virus load and annual increase are strong predictors of disease progression. PMID- 10479127 TI - The effect of drug-injection behavior on genetic evolution of HIV-1. AB - The effect of injection-drug use on human immunodeficiency virus type 1 (HIV-1) env genetic evolution was examined in 15 seroconverting injection-drug users followed up for 4 years. After adjustment for non-drug-related independent variables significantly associated with genetic diversity (time since seroconversion and progressor status), injection frequency was positively and highly significantly associated with HIV-1 env genetic diversity (P=.003). The mutation rate in those who had injected at least once a day during the previous 6 months was estimated to be 62% greater than the rate in those who had not injected at all. If the positive effect of drug-injection frequency on env genetic diversity extends to the HIV-1 pol gene, the risk of emergence of resistance to antiretroviral drugs may be enhanced by increased drug-injection frequency, especially under the selection pressure of antiretroviral therapy. PMID- 10479128 TI - Viral and immunologic examination of human immunodeficiency virus type 1 infected, persistently seronegative persons. AB - Persons who were human immunodeficiency virus type 1 (HIV-1)-infected but who remained persistently seronegative (HIPS) on HIV-1 antibody tests were examined through AIDS case surveillance. Six such individuals (HIPS-1 to -4, -7, and -9) were examined to determine whether their persistent seronegativity was attributable to immune dysfunction or infection with atypical HIV. Of the 6, 4 had antibody titers to at least 1 other common pathogen. In vitro stimulation of peripheral blood mononuclear cells from HIPS-4 and HIPS-7 with pokeweed mitogen or phosphorothioate oligodeoxynucleotide (direct B cell mitogen) did not produce HIV-1-specific antibody. Reconstitution experiments with recombinant interleukin (rIL)-4 and rIL-12 also had no impact on antibody production. Virus isolates from HIPS-4 and -9 were R5X4-tropic, whereas HIPS-7 was CCR5-tropic only. Sequence analysis of long terminal repeat, p24, and env gp41 did not reveal any specific mutation, and phylogenetic analysis confirmed that all 6 virus specimens were HIV 1 subtype B. These data suggest that the lack of a detectable antibody response in these patients may be the result of immune dysfunction. PMID- 10479129 TI - Sequence clusters in human immunodeficiency virus type 1 reverse transcriptase are associated with subsequent virological response to antiretroviral therapy. AB - Many amino acid (aa) sites in reverse transcriptase (RT) have been implicated in resistance to nucleoside (NRTI) and nonnucleoside antiretrovirals. Interactions between these in response to combination therapy remain poorly understood. In a trial (ACTG 241) of zidovudine/didanosine (ddI) versus zidovudine/ddI/nevirapine in nucleoside-experienced patients, baseline sequence data from the RT coding region was analyzed from 55 individuals. Sequences were clustered by use of a parsimony method and the virological responses (ratio of baseline viral load to viral load after of therapy) for each cluster were analyzed at week 8 and week 48. Both clusters and genotype at aa 215 were significantly associated with virological response at both time points, whereas viral load showed a stronger association with sequence clusters. Sequence clusters identified one group of patients who never developed high-level resistance to NRTIs despite prior nucleoside exposure and poor suppression of viral replication. PMID- 10479130 TI - Immune reconstitution after 2 years of successful potent antiretroviral therapy in previously untreated human immunodeficiency virus type 1-infected adults. AB - Today's antiretroviral combination regimens can induce significant and sustained decreases in human immunodeficiency virus (HIV)-RNA levels, allowing the immune system to recover. To what extent immune reconstitution is possible and what factors determine the outcome have thus far not been resolved. We studied 19 subjects, treated for 2 years with protease inhibitor-containing triple therapy, who had a strong suppression of HIV-RNA levels. CD4+ T-cell numbers increased from medians of 170 to 420x106 cells/L, but in a number of subjects T-cell numbers did not further increase after week 72, without having reached normal values. Long-term CD4+ T-cell change was mainly caused by a slow but continuous increase in naive CD4+ T cells (CD45RA+CD62L+) and was predicted by the baseline number of these cells. Our data indicate that long-term immunological recovery is gradual, even during strong suppression of viral replication, not always complete, and dependent on the preexisting level of naive CD4+ T cells. PMID- 10479131 TI - Restoration of normal interleukin-2 production by CD4+ T cells of human immunodeficiency virus-infected patients after 9 months of highly active antiretroviral therapy. AB - The present study investigated immune restoration in patients at intermediate stages of human immunodeficiency virus (HIV) disease after initiation of highly active antiretroviral therapy (HAART). A progressive increase in both memory and naive CD4+ T cells was observed from the first weeks of therapy, concomitant with a decrease in the expression of activation markers on CD8+ T cells. The early activation marker CD69 remained, however, overexpressed on T cells after suboptimal stimulation in vitro, indicative of persistent immune activation. The percentage of interleukin (IL)-2-producing CD4+ T cells significantly increased from 9 months of HAART. In most patients, CD4+ T cells recovered an ability to produce IL-2 on stimulation, similar to that of HIV-seronegative controls. Reversal of T-cell anergy may be a key event in immune restoration for achieving long-term clinical benefit with HAART. PMID- 10479132 TI - The safety and efficacy of granulocyte-macrophage colony-stimulating factor (Sargramostim) added to indinavir- or ritonavir-based antiretroviral therapy: a randomized double-blind, placebo-controlled trial. AB - Sargramostim is a yeast-derived, recombinant human granulocyte-macrophage colony stimulating factor with therapeutic potential in human immunodeficiency virus (HIV) infection. Its safety and activity when used in combination with protease inhibitors were evaluated in a randomized, double-blind trial in which 20 HIV infected subjects on stable antiretroviral regimens, including indinavir or ritonavir, received sargramostim or placebo 3 times a week for 8 weeks. Analysis of HIV virus load excluded any 0. 5 log10 increase due to sargramostim (95% confidence interval, -0.68 to 0.44). Sargramostim was well tolerated, and inflammatory cytokines and surrogate markers of disease progression, such as serum levels of interleukin-10 and soluble tumor necrosis factor receptors types Iota and IotaIota, remained stable in subjects receiving sargramostim. Sargramostim treatment was associated with a trend toward decreased HIV RNA (>0.5 log10) and increased CD4+ cell count (>30%). These results became statistically significant only when subjects with baseline virus loads within the limits of detection or baseline CD4 cell count >50 were analyzed. No difference in indinavir pharmacokinetics was observed before or after sargramostim therapy. PMID- 10479133 TI - Polyspecific self-reactive antibodies in individuals infected with human immunodeficiency virus facilitate T cell deletion and inhibit costimulatory accessory cell function. AB - Self-reactive polyspecific IgG antibodies (PSAs) arise in human immunodeficiency virus (HIV)-seropositive subjects before they develop AIDS. Self-reactive PSA levels correlate with the destruction of CD8 T cells in HIV-infected individuals and mediate the antibody-dependent cellular toxicity-based destruction of human T cells in tissue culture. PSAs react across the species barrier and bind to T cell antigens in mice. Such reactivity with mouse lymphocytes was not detected in normal human serum. Injection of human PSA IgG causes massive T cell depletion in the spleen, lymph nodes, and thymus in mice: evidence that PSA IgG facilitates T cell destruction in vivo. In addition to facilitating macrophage cytotoxicity, self-reactive PSA IgG inhibits the macrophage-mediated activation of T cells with antigen receptor-specific monoclonal antibody or with antigen. Exogenous costimulatory stimuli or interleukin (IL)-12 can reverse the inhibition. In contrast, exogenous IL-10 mimics this inhibition. These data implicate PSA IgG as a pathogenic factor in the development of HIV disease. PMID- 10479134 TI - Immunization with envelope MN rgp120 vaccine in human immunodeficiency virus infected pregnant women. AB - Twenty-six human immunodeficiency virus (HIV)-infected pregnant women participated in a placebo-controlled study of immunogenicity and safety of multiple doses of MN rgp120 vaccine over the last half of pregnancy. The women had CD4 lymphocyte counts>400/mm3, no AIDS-defining illness and normal pregnancies. Vaccination was well tolerated, with no significant local or systemic reactions in the women and no adverse outcomes in the infants attributable to the vaccine. Vaccination did not alter plasma RNA reverse transcriptase-polymerase chain reaction copy number; moreover, immunization was not associated with changes in CD4 counts or HIV binding and neutralization antibody titers. Infants were followed up until 18 months of age. Five of 26 infants (19%) were HIV infected, with infection occurring in children of both vaccinated and placebo women. Analysis of factors that influence transmission did not disclose associations with immunization status, viral load, CD4 count, or maternal viral neutralization titers. PMID- 10479135 TI - Prospective evaluation of amplification-boosted ELISA for heat-denatured p24 antigen for diagnosis and monitoring of pediatric human immunodeficiency virus type 1 infection. AB - The performance in pediatric human immunodeficiency virus type 1 (HIV-1) infection of a signal-amplification boosted ELISA for HIV-1 p24 antigen in plasma after heat-mediated immune complex dissociation was prospectively compared with polymerase chain reaction-based procedures. Diagnostic sensitivity and specificity of the p24 antigen test were 100% and 99.2%, respectively. Quantification revealed RNA in 85.7% and p24 antigen in 87.4% of 230 samples from 25 infected children. Concentrations of these indices in individual samples correlated (P<.0001). Introduction or modification of antiretroviral treatment showed concordant responses of RNA and p24 antigen in 39 (90.7%) of 43 instances. The treatment-induced changes in concentrations of RNA were higher than those of p24 antigen in 11 instances. In 1 instance, however, the concentration change of p24 antigen was greater than that of RNA (P=. 002). Variation of RNA concentrations was more marked than that of p24 antigen (P=.002). The p24 antigen test was equivalent to PCR for diagnosing and monitoring pediatric HIV-1 infection. PMID- 10479136 TI - Chemokine receptor polymorphisms and human immunodeficiency virus disease progression. AB - The role of polymorphisms in genes encoding chemokines and their receptors (CCR2B, SDF-1, and the promoter region of CCR5) in human immunodeficiency virus (HIV) disease progression was studied in 132 white HIV type 1 (HIV-1)-infected participants from a United Kingdom cohort study. Genotyping was done by use of amplification refractory mutation system-polymerase chain reaction with sequence specific primers, and Cox proportional hazards models were used to examine the impact of polymorphisms on time to a CD4 cell count <200x106/L and to CDC stage IV disease. The results confirm a significant association of the CCR2B-64I mutant genotype with slower progression to a CD4 count <200 (hazards ratio [HR], 0.39; 95% confidence interval [CI], 0.17-0.91) but not with the SDF-1alpha 3' UTR homozygous mutation. The effects of the CCR5 and CCR2 mutations were genetically independent and similar in the magnitude of their protective effect on progression to a CD4 count <200 cells. A novel finding was an association of borderline significance between homozygosity for C at nucleotide position 59353 in the CCR5 promoter region and a slower rate of CD4 cell decline to <200x106/L (HR, 0. 58; 95% CI, 0.34-0.996). PMID- 10479137 TI - Adaptation to promiscuous usage of chemokine receptors is not a prerequisite for human immunodeficiency virus type 1 disease progression. AB - Fifty percent of individuals infected with human immunodeficiency virus type 1 (HIV-1) progress to AIDS in the presence of only non-syncytium-inducing (NSI) variants. These rapidly replicating NSI isolates are associated with a high viral load. The question of whether disease progression in the absence of syncytium inducing (SI) HIV-1 variants is associated with an expansion of the coreceptor repertoire of NSI HIV-1 variants was studied. Biological HIV-1 clones were isolated both early and late in infection from progressors and long-term survivors with wild-type or mutant CCR5 or CCR2b genotypes and analyzed for their capacity to use CCR1, CCR2b, CCR3, CCR5, and CXCR4 on U87 cells coexpressing CD4. All HIV-1 clones were restricted to the use of CCR5. Absent replication of all HIV-1 clones in peripheral blood mononuclear cells from a CCR5 Delta32 homozygous blood donor confirmed this result. These findings indicate that an expanded coreceptor repertoire of HIV-1 is not a prerequisite for a progressive clinical course of HIV-1 infection. PMID- 10479138 TI - Lower human immunodeficiency virus (HIV) type 2 viral load reflects the difference in pathogenicity of HIV-1 and HIV-2. AB - Human immunodeficiency virus type 2 (HIV-2) is less pathogenic than HIV type 1 (HIV-1), but the mechanisms underlying this difference have not been defined. We developed an internally controlled quantitative reverse transcriptase-polymerase chain reaction to measure HIV-2 viral load and determined levels of plasma virus in a cohort of registered commercial sex workers in Dakar, Senegal. The assay has a lower limit of detection of 100 copies/mL and is linear over 4 logs. HIV-2 viral RNA was detectable in 56% of all samples tested; the median load was 141 copies/mL. Levels of viral RNA in the plasma were inversely related to CD4+ cell counts. HIV-2 and HIV-1 viral loads were compared among the seroincident women in the cohort; the median viral load was 30x lower in the HIV-2-infected women (P<.001, Wilcoxon rank sum test), irrespective of the length of time infected. This suggests that plasma viremia is linked to the differences in the pathogenicity of the 2 viruses. PMID- 10479139 TI - Efficient human immunodeficiency virus ( HIV)-1 Gag-Env pseudovirion formation elicited from mammalian cells by a canarypox HIV vaccine candidate. AB - Canarypox viruses undergo abortive replication in mammalian cells. Despite this restriction on replication in mammalian cells, significant immune responses have been shown in animals and in humans receiving recombinant canarypox vaccine vectors expressing heterologous immunogens. A recombinant canarypox vaccine candidate (vCP205), which expresses human immunodeficiency virus (HIV)-1 Gag, Env, and protease proteins, is presently under investigation in phase I and phase II human trials in the United States and elsewhere. In this study, the ability of vCP205 to elicit HIV Gag-Env pseudovirion formation in avian and mammalian cells was investigated. Gag-Env pseudovirions were produced from both avian and mammalian cell lines infected by this vaccine vector. A subset of mammalian cells was identified in which pseudovirion production and release was very efficient, surpassing the production from infected avian cells. The production of Gag-Env pseudovirions by canarypox HIV vaccine vectors may have important implications for future HIV vaccine design. PMID- 10479140 TI - Decline of simian immunodeficiency virus (SIV)-specific cytotoxic T lymphocytes in the peripheral blood of long-term nonprogressing macaques infected with SIVmac32H-J5. AB - The evolution of simian immunodeficiency virus (SIV)-specific cytotoxic T lymphocyte precursors (CTLps) and their relationship with virus replication were studied in SIV-infected macaques. After primary viremia, 3 of 8 macaques lost culturable virus and polymerase chain reaction-detectable provirus in peripheral blood. Although proviral DNA persisted in the spleen and lymph nodes, virus loads were below or barely above detection levels. Throughout the study, the 3 macaques remained asymptomatic, with stable CD4+ cell counts. These findings were associated with the detection of CTLps directed against both structural and regulatory SIV proteins. The response peaked during the first 7 months of infection but waned subsequently. CTLps increased after rechallenge of 1 macaque, suggesting that limited antigenic stimulation contributed to their disappearance from circulation. Transient viremia with increasing CTLp frequencies and antibody titers also suggested at least partial susceptibility to reinfection. These findings bear implications for vaccination strategies aimed at inducing protective CTLs against lentiviruses. PMID- 10479141 TI - Involvement of matrix metalloproteinases in human immunodeficiency virus type 1 induced replication by clinical Mycobacterium avium isolates. AB - The role of Mycobacterium avium isolates in modulating human immunodeficiency virus type 1 (HIV-1) replication was examined by use of an in vitro, resting T cell system. Two human clinical isolates (serotypes 1 and 4) but not an environmental M. avium isolate (serotype 2) enhanced HIV-1 replication. The M. avium-induced HIV-1 replication was not associated with cell activation or differential cytokine production or utilization. Addition of matrix metalloproteinase (MMP) inhibitors and their in vivo regulators, tissue inhibitors of metalloproteinases-1 and -2, abrogated M. avium-induced HIV-1 replication 80%-95%. The MMP inhibitors did not have any effect on the HIV-1 protease activity, suggesting that they may affect cellular processes. Furthermore, MMP-9 protein was differentially expressed after infection with clinical M. avium isolates and paralleled HIV-1 p24 production. Collectively, these data suggest that M. avium-induced HIV-1 replication is mediated, in part, through the induction of MMP-9. PMID- 10479142 TI - Decreased point prevalence of Haemophilus influenzae type b (Hib) oropharyngeal colonization by mass immunization of Brazilian children less than 5 years old with hib polyribosylribitol phosphate polysaccharide-tetanus toxoid conjugate vaccine in combination with diphtheria-tetanus toxoids-pertussis vaccine. AB - A protective herd effect has been described after susceptible populations of children are vaccinated with conjugate Haemophilus influenzae type b (Hib). Hib carriage was studied in children aged 6-24 months attending day care centers in two cities in southern Brazil (Curitiba and Porto Alegre). In Curitiba, routine immunization with Hib polyribosylribitol phosphate polysaccharide-tetanus toxoid conjugate vaccine (PRP-T) in combination with diphtheria-tetanus toxoids pertussis vaccine (PRP-T/DTP) has been offered since September 1996; DTP vaccine alone is routinely given in Porto Alegre. Children in Porto Alegre (n=643) were 8 times less likely to have received adequate Hib vaccination and 4 times more likely to be Hib carriers than children in Curitiba (n=647; i.e., point prevalence of oropharyngeal colonization, 4.8% vs. 1.2%). Point prevalence of carriage with non-type b or other nontypeable Hi was similar in children of both cities. There was a vaccination effect on carriage rates in children who received a primary 3-dose series, independent of the booster dose, suggesting that a booster may be unnecessary to induce population protection. PMID- 10479143 TI - Incident syphilis among women with multiple admissions to jail in New York City. AB - Although early syphilis morbidity in New York City (NYC) has declined to a record low, syphilis seroreactivity among women jailed in NYC is approximately 25%. By use of a retrospective cohort-type analysis of longitudinal serologic and treatment data collected at the time of each incarceration, the incidence of syphilis infection among 3579 susceptible women jailed multiple times in NYC between 23 March 1993 and 10 April 1997 was estimated. Syphilis incidence densities were estimated by use of continuous, time-homogeneous Markov models. There was a total of 289 incident infections. The overall incidence density was 6.5 infections per 100 woman-years (95% confidence interval, 5.7-7.2), which exceeds the 1997 early syphilis rate among women in NYC by>1000-fold. The persisting high incidence of syphilis in this population underscores the importance of aggressive syphilis control in correctional settings, even in the face of declining local early syphilis rates. PMID- 10479144 TI - Role of peroxynitrite as a mediator of pathophysiological alterations in experimental pneumococcal meningitis. AB - This study investigated the role of peroxynitrite in an adult rat model of pneumococcal meningitis. Immunohistochemically, nitrotyrosine residues, as a marker for peroxynitrite formation, were detected perivascularly and in proximity to inflammatory cells in the subarachnoid space. Nitrotyrosine immunoreactivity was colocalized with blood-brain barrier breaching, which was visualized by fluorescence microscopy after intravenous application of Evans blue. Treatment of infected rats with uric acid (300 mg/kg intraperitoneally), a scavenger of peroxynitrite, significantly attenuated intracranial pressure, cerebrospinal fluid white blood cell count, and blood-brain barrier leakage, as indicated by Evans blue concentration in the cerebrospinal fluid (21.6+/-9.3 mm Hg, 5776+/ 1790 cells/microL, 9.7+/-6.4 microgram/mL in infected, untreated rats vs. 7.2+/ 1.6 mm Hg, 2004+/-904 cells/microL, 1.1+/-1.0 microgram/mL infected, uric acid treated rats, mean+/-SD, P<.05). These data suggest that peroxynitrite plays a central role in mediating pathophysiological alterations during bacterial meningitis. PMID- 10479145 TI - Immunogenicity and impact on nasopharyngeal carriage of a nonavalent pneumococcal conjugate vaccine. AB - The safety, immunogenicity, and impact on carriage of a nonvalent pneumococcal vaccine given at ages 6, 10, and 14 weeks were examined in a double-blind, randomized, placebo-controlled trial in 500 infants in Soweto, South Africa. No serious local or systemic side effects were recorded. Significant antibody responses to all pneumococcal serotypes were observed 4 weeks after the third dose. Haemophilus influenzae type b polyribosylribitol phosphate (geometric mean titer, 11.62 microg/mL) and diphtheria (1.39 IU/mL) antibodies were significantly higher in children receiving pneumococcal conjugate, compared with placebo recipients (4.58 microgram/mL and 0.98 IU/mL, respectively). Nasopharyngeal carriage of vaccine serotypes decreased in vaccinees at age 9 months (18% vs. 36%), whereas carriage of nonvaccine serotypes increased (36% vs. 25%). Carriage of penicillin-resistant pneumococci (21% vs. 41%) and cotrimoxazole-resistant pneumococci (23% vs. 35%) were significantly reduced 9 months after vaccination, compared with controls. PMID- 10479146 TI - A cluster of VanD vancomycin-resistant Enterococcus faecium: molecular characterization and clinical epidemiology. AB - VanD-mediated glycopeptide resistance has been reported for an isolate of Enterococcus faecium, BM4339. Three clinical isolates of vancomycin-resistant E. faecium collected from 3 patients during a 6-week period in 1993 had agar dilution MICs of vancomycin and teicoplanin of 128 and 4 microg/mL, respectively. Polymerase chain reaction (PCR) using degenerate primers complementary to genes encoding d-Ala-d-X ligases yielded a 630-bp product that was similar to the published partial sequence of vanD. By use of inverse PCR, vanD, vanHD, and two partial flanking open-reading frames were sequenced. The deduced amino acid sequence of VanD showed 67% identity with VanA and VanB. vanD appeared to be located on the chromosome and was not transferable to other enterococci. The 3 isolates were indistinguishable by pulsed-field gel electrophoresis and differed from BM4339. No other isolates carrying vanD were found in a subset of 875 recent US isolates of vancomycin-resistant enterococci. PMID- 10479147 TI - Suppression of acquired immunity against Listeria monocytogenes by amphotericin B mediated inhibition of CD8 T cell function. AB - Amphotericin B is frequently used for the treatment of fungal infections of immunocompromised individuals. Whereas immunomodulatory side effects of this agent are known, the influence of amphotericin B was studied in the model of murine Listeria monocytogenes infection. Treatment of L. monocytogenes-immune mice with a nontoxic dose of amphotericin B (0.75 mg/kg) reduced antilisterial protection by 4-5 orders of magnitude, while it had no significant effect on natural immunity against L. monocytogenes in naive mice. Treatment of mice with amphotericin B also abolished the protection mediated by transfer of an L. monocytogenes-specific CD8 T cell line. Furthermore, in vitro analysis showed that amphotericin B impaired target cell lysis and interferon-gamma production by peptide-specific CD8 T cell lines and antigen presentation by L. monocytogenes infected macrophagelike cells. These data indicate that amphotericin B has a strong suppressive effect on the function of CD8 T cells in vitro and in vivo. PMID- 10479148 TI - Modulation of intracellular growth of Listeria monocytogenes in human enterocyte Caco-2 cells by interferon-gamma and interleukin-6: role of nitric oxide and cooperation with antibiotics. AB - The influence of interferon (IFN)-gamma and interleukin (IL)-6 on the intracellular growth of Listeria monocytogenes phagocytosed from the apical pole was examined in polarized Caco-2 cells. IFN-gamma (from the apical pole) and IL-6 (from the basolateral pole) considerably reduced the bacterial intracellular growth, an effect largely abolished by l-monomethyl arginine. Both cytokines caused overexpression of inducible nitric oxide synthase. IL-6, but not IFN gamma, caused a partial restriction of L. monocytogenes in phagosomes and largely prevented the cytosolic forms from being surrounded by actin. Ampicillin was bacteriostatic in unstimulated cells but modestly bactericidal in cells treated with IFN-gamma and IL-6. Azithromycin (a macrolide) was fairly bactericidal and sparfloxacin (a fluoroquinolone) highly bactericidal in all situations. IFN-gamma and IL-6 may therefore be important determinants in the protection of epithelial cells from intracellular multiplication of L. monocytogenes. Ampicillin may fail in their absence, requiring the use of other antibiotics such as the fluoroquinolones. PMID- 10479149 TI - Interaction of Shiga toxins with human brain microvascular endothelial cells: cytokines as sensitizing agents. AB - Neurologic abnormalities are among the most serious extraintestinal complications of infection with Shiga toxin (Stx)-producing bacteria. Histopathologic examination of tissues from patients with extraintestinal sequelae suggested that Stxs damage endothelial cells. It is shown here that human brain microvascular endothelial cells (HBMECs) are relatively resistant to purified Stxs (50% cytotoxic doses [CD50s] >/=10 microgram/mL). Pretreatment of HBMECs with tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, n-butyric acid, or a cAMP analogue resulted in a 103- to 104-fold decrease in CD50 values and a 2- to 4 fold increase in fluoresceinated Stx binding to HBMECs. Treatment of HBMECs with lipopolysaccharides did not significantly alter cytotoxicity or toxin binding. TNF-alpha and IL-1beta treatment was associated with the increased HBMEC expression of the toxin-binding glycolipid globotriaosylceramide. HBMECs did not produce IL-1beta and produced only trace amounts of TNF-alpha when stimulated with purified Stx1 in vitro. PMID- 10479150 TI - An increase in sporadic and outbreak-associated Salmonella enteritidis infections in Wisconsin: the role of eggs. AB - In Wisconsin, reported Salmonella enterica serotype enteritidis (SE) infections during 1997 more than doubled compared with the previous 9 years. A case-control study was conducted to determine risk factors for sporadic infections, and results of outbreak investigations were reviewed. Eating raw eggs (matched odds ratio [MOR]=14.5; 95% confidence interval [CI], 1.7-591.6), eating raw or undercooked eggs (MOR=5.8; 95% CI, 1.3-28.0), eating any eggs (MOR=4.2; 95% CI, 1.2-16.2), and dining at a restaurant (MOR=4.7; 95% CI, 1.4-18.4) were associated with infection in the case-control study. For 3 of the 8 outbreaks, a probable source was identified, in each instance, foods containing eggs. Human infections decreased after eggs were diverted from implicated flocks. This epidemic demonstrates the continuing need for quality assurance on egg farms and enhanced education of consumers and commercial food preparers regarding safe handling of eggs. PMID- 10479151 TI - Neutrophil recruitment and resistance to urinary tract infection. AB - This study examined the role of neutrophil leukocytes for the antibacterial defense at mucosal infection sites. Urinary tract infection (UTI) was established by injection into the bladder lumen of Escherichia coli 1177, a fully virulent clinical isolate. Infection of C3H/HeN (lpsn, lpsn) mice recruited neutrophils into the urinary tract, and bacteria were cleared from kidneys and bladders. The neutrophil response was absent in C3H/HeJ (lpsd, lpsd) mice, and bacteria persisted in the tissues. Peripheral neutrophil depletion of C3H/HeN mice was subsequently achieved by pretreatment with the granulocyte-specific antibody RB6 8C5. The E. coli-induced neutrophil recruitment was inhibited, as shown by immunohistochemistry and tissue myeloperoxidase quantitation. As a consequence, bacterial clearance from kidneys and bladders was drastically impaired. Antibody treatment of C3H/HeJ mice had only a marginal effect. The results show that neutrophils are essential for bacterial clearance from the urinary tract and that the neutrophil recruitment deficiency in C3H/HeJ mice explains their susceptibility to gram-negative UTI. PMID- 10479152 TI - Interleukin 10 produced by macrophages inoculated with Mycobacterium avium attenuates mycobacteria-induced apoptosis by reduction of TNF-alpha activity. AB - Normal human macrophages respond to infection with Mycobacterium avium, serovar 4, by producing tumor necrosis factor (TNF)-alpha, which mediates apoptosis, and by elaborating interleukin (IL)-10, a TNF-alpha antagonist. We show that IL-10 down-regulates apoptosis by inhibiting the TNF-alpha production of the inoculated macrophages and by inducing the release of soluble TNF receptor type 2 from the macrophages, which leads to inactivation of TNF-alpha. These experiments suggest that induction of IL-10 production is a virulence factor that creates an intracellular sanctuary for the bacteria that is inaccessible to the defense mechanisms of the host. PMID- 10479153 TI - Analysis of rate of change of IS6110 RFLP patterns of Mycobacterium tuberculosis based on serial patient isolates. AB - The rate of change of IS6110 restriction fragment length polymorphism (RFLP) patterns of Mycobacterium tuberculosis was determined in serial isolates from 544 patients. In 25 patients (4.6%), the RFLP patterns of the follow-up isolates differed from the initial isolates. Patients with different follow-up strains were less likely to cluster with patients whose strains had indistinguishable RFLP patterns. Changes in RFLP patterns were more common for persons with extrapulmonary disease and for those who had both pulmonary and extrapulmonary isolates. Based on serial isolates spanning for the most part <3 months, the half life was extrapolated to be 3.2 years (95% confidence interval, 2.1-5.0). The main implication of this study is that the rate of change of IS6110-based RFLP of M. tuberculosis supports the use of IS6110 typing in epidemiologic studies of recent transmission of tuberculosis. PMID- 10479154 TI - Complex transmission dynamics of clonally related virulent Mycobacterium tuberculosis associated with barhopping by predominantly human immunodeficiency virus-positive gay men. AB - Limited data suggest that measures to reduce tuberculosis transmission should be based on locations rather than on personal contacts. Molecular epidemiologic methods (analysis of IS6110 patterns, spoligotypes, variable numbers of tandem DNA repeats, and automated DNA sequence data) identified a cohort of 48 persons who were infected with progeny of the same Mycobacterium tuberculosis strain. Epidemiologic investigation documented that a large proportion of the patients were gay white human immunodeficiency virus-positive men. Most practiced barhopping, an activity that involved patronizing many bars in the same neighborhood each night. Few subjects were directly linked to more than 1 or 2 other persons by conventional investigation methods, which shows that the transmission dynamics were unusually complex compared with most previously described episodes of strain spread. The data support the concept that identification of locations where pathogen dissemination likely occurs may provide additional strategies for targeted tuberculosis control. PMID- 10479155 TI - The effect of doxycycline treatment on the development of protective immunity in a murine model of chlamydial genital infection. AB - Chlamydia trachomatis is a major cause of sexually transmitted disease (STD) worldwide. Antibiotics are effective in treating infection; however, reinfection is common. This observation has led to the conclusion that infection fails to elicit a protective antichlamydial immune response. It was postulated that high reinfection rates might be due to early eradication of organisms from genital tissue after antibiotic intervention, which could negatively influence the development of naturally acquired protective immunity. This hypothesis was tested by use of a murine model of female genital infection. The findings show that doxycycline intervention of infection, although very effective in eradicating chlamydiae from genital tissue and preventing upper genital tract disease, significantly inhibits the development of protective immunity. If antibiotic intervention of human chlamydial genital infection has a similar effect on protective immunity, it could have important implications in the understanding of immunity to infection and future public health efforts to control chlamydial STD. PMID- 10479156 TI - Amphotericin B-induced interleukin-1beta expression in human monocytic cells is calcium and calmodulin dependent. AB - Amphotericin B remains the agent of choice for treatment of severe fungal infections. Its use is hindered by adverse effects, including infusion-related fever, chills, and hypotension, as well as nephrotoxicity with secondary anemia, hypokalemia, and hypomagnesemia. Amphotericin B-induced transcription and expression of interleukin (IL)-1beta by human monocytes is believed to be involved in mediating infusion-related adverse effects. It is shown here that agents that increase intracellular calcium [Ca++]i (A23187 and thapsigargin) in human monocytic cells also induce IL-1beta expression. Furthermore, amphotericin B-induced IL-1beta expression is attenuated by the calmodulin antagonist calmidazolium. Amphotericin B 5.41 microM increases [Ca++]i by up to 300 nM in these cells. In the presence of a nominal calcium buffer or EGTA, amphotericin B induced IL-1beta expression is attenuated. Thus, amphotericin B acts as an ionophore to increase [Ca++]i and activates calmodulin-mediated expression of IL 1beta in human monocytes. PMID- 10479157 TI - Proteases from Aspergillus fumigatus induce interleukin (IL)-6 and IL-8 production in airway epithelial cell lines by transcriptional mechanisms. AB - Proteases secreted by Aspergillus fumigatus induce the production of cytokines by epithelial cells, including interleukin (IL)-6 and IL-8. In the present study, we focused on the mechanism(s) by which A. fumigatus-derived proteases elicit cytokine production in epithelial cells. In the epithelial cell line A549, IL-6 and IL-8 mRNA levels were enhanced by proteases as a result of transcriptional induction of the respective genes. Transcriptional induction of both genes coincided with enhanced DNA binding of nuclear factor (NF)-kappaB and NF-IL6, whereas activator protein-1 was unlikely to be involved. The enhanced transcriptional activity could be inhibited by the addition of chymostatin, showing serine protease dependency. Posttranscriptional mechanisms affecting the stability of IL-6 and IL-8 mRNAs were not involved in protease-induced IL-6 and IL-8 production. These data show that after exposure to A. fumigatus-derived proteases, IL-6 and IL-8 gene expressions are up-regulated as a result of transcriptional mechanisms. PMID- 10479158 TI - Virulence of three distinct Cryptosporidium parvum isolates for healthy adults. AB - The infectivity of three Cryptosporidium parvum isolates (Iowa [calf], UCP [calf], and TAMU [horse]) of the C genotype was investigated in healthy adults. After exposure, volunteers recorded the number and form of stools passed and symptoms experienced. Oocyst excretion was assessed by immunofluorescence. The ID50 differed among isolates: Iowa, 87 (SE, 19; 95% confidence interval [CI], 48.67-126); UCP, 1042 (SE, 1000; 95% CI, 0-3004); and TAMU, 9 oocysts (SE, 2.34; 95% CI, 4.46-13.65); TAMU versus Iowa, P=.002 or UCP, P=.019. Isolates also differed significantly (P=.045) in attack rate between TAMU (86%) and Iowa (52%) or UCP (59%). A trend toward a longer duration of diarrhea was seen for the TAMU (94.5 h) versus UCP (81.6 h) and Iowa (64.2 h) isolates. C. parvum isolates of the C genotype differ in their infectivity for humans. PMID- 10479159 TI - Randomized dose-ranging study of the safety and efficacy of WR 238605 (Tafenoquine) in the prevention of relapse of Plasmodium vivax malaria in Thailand. AB - WR 238605 is an 8-aminoquinoline developed for the radical cure of Plasmodium vivax. Forty-four P. vivax-infected patients were randomly assigned to 1 of 4 treatment regimens: 3 groups received a blood schizonticidal dose of chloroquine followed by WR 238605: group A (n=15) received 300 mg daily for 7 days; group B (n=11), 500 mg daily for 3 days, repeated 1 week after the initial dose; group C (n=9), 1 dose of 500 mg. A fourth group (D; n=9) received chloroquine only. Among patients who completed 2-6 months of follow-up (n=23), there was 1 relapse in group B (day 120) and 1 in group C (day 112). Among patients treated with chloroquine only, there were 4 relapses (days 40, 43, 49, and 84). WR 238605 was safe, well tolerated, and effective in preventing P. vivax relapse. PMID- 10479160 TI - The prognostic and pathophysiologic role of pro- and antiinflammatory cytokines in severe malaria. AB - Pro- and antiinflammatory cytokines were measured on admission in 287 consecutive Vietnamese adults with severe falciparum malaria. Plasma interleukin (IL)-6, IL 10, and tumor necrosis factor (TNF)-alpha concentrations and the IL-6: IL-10 ratio were significantly higher in patients who died than in survivors (P<.001). On multivariate analysis, hyperparasitemia, jaundice, and shock were all associated independently with raised IL-6, IL-10, and interferon-gamma, and acute renal failure specifically with raised TNF-alpha levels. Cerebral malaria patients, particularly those without other vital organ dysfunction, had significantly lower levels of these cytokines (P=.006), reflecting a more localized pathology. Serial IL-6 and IL-10 measurements made on 43 patients who died and matched survivors indicated a relative deficiency in IL-10 production as death approached. Elevated plasma cytokines in severe malaria are associated with systemic pathologic abnormalities, not cerebral involvement. Both the overall magnitude of the cytokine responses and the eventual imbalance between the pro- and antiinflammatory responses are important determinants of mortality. PMID- 10479161 TI - Susceptibility to periportal (Symmers) fibrosis in human schistosoma mansoni infections: evidence that intensity and duration of infection, gender, and inherited factors are critical in disease progression. AB - Lethal disease in Schistosoma mansoni infections is mostly due to portal hypertension caused by hepatic periportal fibrosis. To evaluate the factors that may determine severe disease, livers and spleens were examined by ultrasound in a Sudanese population living in a village where S. mansoni is endemic. Early (FI), moderate (FII), or advanced (FIII) fibrosis was observed in 58%, 9%, and 3% of the population, respectively. Although FI affected 50%-70% of the children and adolescents, FII prevalence was low in subjects median) was associated with improved survival (P=.05). A similar trend was observed for NK activity (P=.1). In a multivariate model controlling for baseline CD4 cell count, HIV RNA, and use of protease inhibitors during follow-up, high ADCC, but not high NK activity, remained significantly associated with a lower risk of death (relative risk, 0.18; 95% confidence interval, 0.05-0.75). ADCC may be an important determinant of disease progression independently of anti retroviral therapy, CD4 cell count, and HIV RNA. PMID- 10479167 TI - Hepatitis G virus and human immunodeficiency virus coinfection: response to interferon-alpha therapy. AB - The prevalence and consequences of hepatitis G virus (HGV) infection were determined in 180 patients with human immunodeficiency virus (HIV) infection (predominantly male homosexuals) who participated in a trial that compared treatment with zidovudine versus interferon (IFN)-alpha versus the combination. HGV RNA levels were measured by branched DNA signal amplification assay. Initially, 66 (37%) had HGV RNA. Sexual transmission was the sole risk factor for infection in all but 4 subjects. Pretreatment clinical features were similar between HGV RNA-positive and -negative patients. After 6 months, only 5% treated with zidovudine became HGV RNA negative, compared with 95% who received IFN-alpha alone and 66% on combination therapy with low-dose IFN-alpha. After therapy, HGV RNA levels returned to baseline in most subjects. Thus, HGV infection is common among HIV-infected homosexual males but does not appear to influence clinical features in early HIV infection. HGV RNA levels are suppressed by IFN but not by zidovudine. PMID- 10479169 TI - Highly active antiretroviral treatment initiated early in the course of symptomatic primary HIV-1 infection: results of the ANRS 053 trial. AB - Highly active antiretroviral treatment (HAART) was given early to 64 patients with symptomatic primary human immunodeficiency virus (HIV)-1 infection. At the time of analysis, patients had been followed up for 9-21 months. No patient had died or developed an AIDS-defining event. Survival analysis showed that by month 21 the proportion of patients with plasma HIV-1 RNA <50 copies/mL was 72% (95% confidence interval, 58%-95%) in intention-to-treat analysis. After 18 months of treatment, 50% of the patients with undetectable plasma HIV-1 RNA also had undetectable HIV-1 RNA in peripheral blood mononuclear cells (PBMC). Only 1 of 3 patients had undetectable HIV-1 RNA in lymphoid tissue, while all patients had quantifiable HIV-1 DNA both in PBMC and lymphoid tissue. The median CD4 lymphocyte increase from baseline was 230 cells/microL. These preliminary results support the use of HAART in patients with primary HIV-1 infection. PMID- 10479170 TI - Suppression of plasma virus load below the detection limit of a human immunodeficiency virus kit is associated with longer virologic response than suppression below the limit of quantitation. AB - Suppression of human immunodeficiency virus type 1 plasma virus load (PVL) to <20 copies/mL is associated with a longer virologic response after initiation of antiretroviral therapy. The relationship between duration of virologic response and PVL nadir according to a less sensitive assay was explored. When compared with subjects with a PVL nadir >500 copies/mL, the relative risks of PVL rising above 1000 copies/mL for participants in the INCAS trial and the British Columbia Drug Treatment Program with a PVL nadir below the limit of detection (LOD) were 0.04 (95% confidence interval [CI], 0.02-0.09) and 0.06 (95% CI, 0.03-0.12), respectively. The corresponding relative risks for persons with a detectable but not quantifiable PVL nadir were 0.25 (95% CI, 0.13-0.50) and 0.54 (95% CI, 0.25 1.19). The relative risks of virologic failure associated with a PVL nadir detectable but not quantifiable and a PVL nadir below the LOD were statistically different (P<.0001) in both data sets. PMID- 10479171 TI - Systemic and mucosal immunity is elicited after both intramuscular and intravaginal delivery of human immunodeficiency virus type 1 DNA plasmid vaccines to pregnant chimpanzees. AB - DNA vaccines encoding human immunodeficiency virus type 1 (HIV-1) env/rev and gag/pol were delivered intravaginally (IVAG) and intramuscularly (IM) to 2 pregnant chimpanzees. Vaccination was well tolerated and each chimpanzee developed antibodies (up to 1 year later) to both vaccines. Placental transfer of anti-Env and anti-Gag IgG was demonstrated in both maternal/infant pairs. Specific IgG was also demonstrated in saliva, vaginal, and rectal washes after IVAG immunization. Predominantly anti-HIV-1 IgA was detected in the milk of both mothers after both IM and IVAG immunization. Cellular responses included Gag specific proliferation of lymphocytes and cytotoxic T lymphocytes against both antigens. These data suggest a strategy for induction of mucosal and systemic responses after both IM and IVAG delivery of DNA vaccines in a primate model and could ultimately be useful in lowering maternal-to-fetal transmission of HIV-1, perinatally and through breastfeeding. PMID- 10479172 TI - The effect of age and natural priming on the IgG and IgA subclass responses after parenteral influenza vaccination. AB - This study investigated the effect of natural priming and age on serum IgG and IgA subclass responses after parenteral trivalent influenza vaccination. Sera from 18 young children and 8 adults were collected at various times after vaccination. An ELISA was performed to quantify the concentrations of antibody subclasses. The children were divided into primed and unprimed groups based on the presence of prevaccination serum antibodies. In both children and adults, IgG1 and IgA1 were the predominant IgG and IgA subclasses detected after vaccination. No IgG2 responses were detected in sera of unprimed children, and the proportion of the IgG2 response was lower in primed children than in adults. This suggests that the IgG2 immune response in young children is dependent on previous priming and may mature later than the other IgG subclasses after parenteral influenza vaccination. PMID- 10479173 TI - An outbreak of Salmonella serogroup Saphra due to cantaloupes from Mexico. AB - An outbreak of Salmonella serogroup Saphra (S. saphra) infections was studied by laboratory-based surveillance, case-control and trace-back studies, and a survey of cantaloupe preparation practices. Twenty-four patients with S. saphra infections had illness onsets between 23 February and 15 May 1997; 75% were 1 microg/mL postprimary series (avidity index [AI], 42%, 95% confidence interval [CI], 35% 49%, and 68% and 63%-72%, respectively; P<.0001). For infants with <1 microgram/mL anti-Hib-PS IgG antibody, mean AI rose by the time of preboost immunization to 61% (95% CI, 57%-65%), even though total IgG antibody levels fell. Spontaneous Hib-PS antibody rises after primary series DTaP-PRP-T-HB vaccination were followed by high postboost anti-Hib-PS IgG antibody levels and avidity. The DTaP-PRP-T-HB combination vaccine studied elicits high avidity antibody, and affinity maturation appears to occur in the absence of further antigen exposure even in those with very low anti-Hib-PS antibody. PMID- 10479181 TI - Eotaxin expression in Onchocerca volvulus-induced dermatitis after topical application of diethylcarbamazine. AB - In persons with onchocerciasis, topical application of the anthelminthic diethylcarbamazine (DEC) induces clinical and histologic responses similar to acute papular onchodermatitis, including recruitment of eosinophils to the skin. To determine whether the eosinophil chemokine eotaxin is likely to be associated with eosinophil recruitment in onchodermatitis, DEC was applied to a 5-cm2 area on the skin of infected persons, and biopsies were taken from lesions 24 h later. Histologic analysis showed elevated dermal and epidermal eosinophils compared with tissue from an adjacent (untreated) site. Reverse transcription-polymerase chain reaction showed that eotaxin gene expression in DEC-treated skin was elevated 2- to 17-fold compared with control tissue. Eotaxin immunoreactivity was noted in mononuclear cells and eosinophils in the perivascular region of the dermis and in lymphatic and vascular endothelial cells. Together, these observations are consistent with a role for eotaxin in recruitment of eosinophils to the dermis in early stage onchocercal skin disease. PMID- 10479182 TI - Trans-sialidase from Trypanosoma cruzi induces apoptosis in cells from the immune system in vivo. AB - Trypanosoma cruzi, the etiologic agent of Chagas' disease, expresses trans sialidase, an enzyme able to direct transfer of sialyl residues among macromolecules. The enzyme is shed and can be detected in blood during the acute phase of the disease. Several alterations of the immune response and apoptosis of cellular components of the immune system are observed early in the infection. The possible involvement of bloodstream trans-sialidase on these events was analyzed here. The enzyme induced apoptosis in cells of the immune system in the spleen, thymus, and peripheral ganglia. Both natural and recombinant trans-sialidases induced apoptosis to a similar extent. No effect was detected when enzymatically inactive recombinant molecules were used. In dose-response assays, apoptosis was observed even when an amount of trans-sialidase was administered that was enzymatically undetectable in blood. These findings strongly suggest a role for sialic acid mobilization in T. cruzi-induced apoptosis of immune system cells. PMID- 10479183 TI - Long-term suppression of Mansonella streptocerca microfilariae after treatment with ivermectin. AB - The long-term effect of a single oral dose of 150 microg/kg of body weight of ivermectin on Mansonella streptocerca microfilariae was studied in western Uganda. Before treatment, the geometric mean microfilaria density (mf) in 93 infected persons was 2.4 mf/mg of skin (range, 0.1-42.6). One year after treatment, 43 persons (46%) were microfilaria-negative, and the geometric mean in the remaining persons dropped significantly, to 0.7 mf/mg (range, 0.1-6.9). Thus, ivermectin is highly effective against M. streptocerca, and a single dose leads to a sustained suppression of microfilariae in skin. In Africa, ivermectin is used for mass treatment to control Onchocerca volvulus and Wuchereria bancrofti. Because these filarial parasites are often coendemic with M. streptocerca, the treated population may receive the additional benefit of suppression of M. streptocerca microfilariae. PMID- 10479184 TI - Cytotoxic T lymphocyte precursors in persons with repeated exposure to human immunodeficiency virus. PMID- 10479185 TI - Paternal and maternal infection status and Helicobacter pylori in their children. PMID- 10479187 TI - Reply. PMID- 10479186 TI - Modulation of the pro- and antiinflammatory cytokines by amphotericin B. PMID- 10479188 TI - Influence of hepatitis C virus (HCV) genotype, HCV RNA load, and alanine aminotransferase level on reduction of HCV RNA after a single administration of interferon-alpha. PMID- 10479189 TI - Cryptococcosis and smoking: the potential role of iron. PMID- 10479191 TI - Whole grains and protection against coronary heart disease: what are the active components and mechanisms? PMID- 10479190 TI - Mechanisms regulating leptin production: implications for control of energy balance. PMID- 10479192 TI - Potential interventions for the prevention of childhood pneumonia in developing countries: improving nutrition. AB - Acute respiratory infections are the leading cause of childhood death in developing countries. Current efforts at mortality control focus on case management and immunization, but other preventive strategies may have a broader and more sustainable effect. This review, commissioned by the World Health Organization, examines the relations between pneumonia and nutritional factors and estimates the potential effect of nutritional interventions. Low birth weight, malnutrition (as assessed through anthropometry), and lack of breast feeding appear to be important risk factors for childhood pneumonia, and nutritional interventions may have a sizeable effect in reducing deaths from pneumonia. For all regions except Latin America, interventions to prevent malnutrition and low birth weight look more promising than does breast-feeding promotion. In Latin America, breast-feeding promotion would have an effect similar to that of improving birth weights, whereas interventions to prevent malnutrition are likely to have less of an effect. These findings emphasize the need for tailoring interventions to specific national and even local conditions. PMID- 10479193 TI - Effect of prolonged moderate and severe energy restriction and refeeding on plasma leptin concentrations in obese women. AB - BACKGROUND: Plasma leptin in humans is subject to both long- and short-term regulation; it correlates with indexes of body fat that can only change slowly. However, short-term fasting causes large and rapid decreases. OBJECTIVE: We tested the interactions between energy intake and fat loss on plasma leptin during prolonged moderate and severe energy restriction, with a view to understanding mechanisms of control. DESIGN: Postabsorptive leptin was measured with an enzyme-linked immunosorbent assay specific for the human peptide in 21 obese women aged 41 +/- 3 y (weight: 102 +/- 4 kg; 48 +/- 1% body fat) after 1 wk of a weight-maintaining diet and then weekly for 4 wk during a total fast (group 1); a 1.9-MJ/d all-protein, very-low-energy diet (VLED) (group 2); or a low energy, balanced-deficit diet (BDD) providing 50% of maintenance energy (group 3). In groups 1 and 2, leptin was also measured after 1 wk of refeeding with a diet equivalent to the BDD. RESULTS: Mean leptin decreased markedly by up to 66% (P < 0.001) at week 1 of energy restriction and then gradually thereafter. The change in leptin per kilogram fat mass correlated with that in glucose concentrations [r = 0.538 (P = 0.012) at week 1 and r = 0.447 (P = 0.042) at week 4] but not with that in fat mass. During refeeding postfasting, leptin increased (P = 0.008), despite an ongoing loss of fat mass and correlated positively with changes in resting energy expenditure. At times with comparable cumulative energy restriction and fat loss between diets, the percentage change in leptin paralleled that in glucose. CONCLUSIONS: In obesity, changes in energy intake over days to weeks are a primary modulator of plasma leptin concentrations that are related to the change in glycemia and are able to override the regulatory influence of fat mass. PMID- 10479194 TI - Heart rate variability and fatty acid content of blood cell membranes: a dose response study with n-3 fatty acids. AB - BACKGROUND: Dietary intake of long-chain n-3 polyunsaturated fatty acids (PUFA) may protect against sudden cardiac death, an event that may be predicted by measurement of heart rate variability (HRV). OBJECTIVE: The objectives of this study were to 1) examine the correlations between the content of fatty acids in blood cell membranes (platelets and granulocytes) and HRV in healthy subjects, and 2) assess the effect on HRV of dietary intervention with n-3 PUFA in different doses. DESIGN: Sixty healthy volunteers (25 women and 35 men) were randomly assigned to 3 treatment groups in a double-blind design. Subjects received a daily supplement of either 6.6 g n-3 PUFA, 2.0 g n-3 PUFA, or placebo (olive oil). A 24-h Holter recording was obtained for each subject before supplementation and after 12 wk of supplementation; the 24-h HRV was then related to the content of fatty acids in granulocytes and platelets. RESULTS: Before supplementation, positive correlations were observed in men between the content of docosahexaenoic acid in cell membranes and HRV indexes (r = 0.50, P < 0.01), whereas such correlations were not found in women. Dietary intervention revealed a dose-dependent effect of n-3 PUFA on HRV in men, whereas no effect was found in women. CONCLUSION: The study showed a beneficial effect of n-3 PUFA on HRV in healthy men, suggesting an antiarrhythmic effect of n-3 PUFA. No such effect was observed in healthy women. PMID- 10479195 TI - Long-term efficacy and safety of a new olive oil-based intravenous fat emulsion in pediatric patients: a double-blind randomized study. AB - BACKGROUND: A new intravenous lipid emulsion (ILE) prepared from a mixture of soybean and olive oils contains only long-chain triacylglycerols, with a low proportion (20%) of polyunsaturated fatty acids and 60% monounsaturated fatty acids. OBJECTIVE: The goal of this randomized, double-blind clinical trial was to assess in children the efficacy and safety of this new ILE compared with a control group receiving a soybean-oil emulsion. DESIGN: Eighteen children received for 2 mo 24% of nonprotein energy (1.80 g kg (-)(1) d(-)(1)) either as the new ILE or a soybean oil-based emulsion. Assessments were performed on days 30, 0, 30, and 60 and the changes (day 60 - day 0) assessed by analysis of variance. RESULTS: There were no significant differences in triacylglycerol, apolipoproteins A-I and B, or HDL cholesterol between the 2 groups, whereas total and LDL cholesterol were higher in the soybean oil group on day 60. The pattern of 20:4n-6 in erythrocyte membranes did not change significantly, nor did the ratio of 20:3n-9 to 20:4n-6. On day 60, 18:1n-9 was significantly higher in the olive oil group, the ratio of Sigma(n)-6 > C(18) + 18:3n-6 to 18:2n-6 was 2.20 +/ 0.09 in the olive oil group and 1.33 +/- 0.16 in the soybean-oil group, and Sigma(n)-3 > C(18) was 3.83 +/- 0.30 in the olive oil group and 4. 03 +/- 0.33 in the soybean-oil group. The peroxidation index was lower after the olive oil treatment. CONCLUSIONS: The olive oil-based emulsion was well tolerated, maintained a normal EFA status, and may be more suitable for prevention of lipid peroxidation than the soybean-oil-based emulsion. PMID- 10479196 TI - Alterations in body weight and composition consequent to 20 wk of endurance training: the HERITAGE Family Study. AB - BACKGROUND: Obesity is a major public health problem in the United States. The role of physical activity and formal exercise in controlling body weight has not been clearly determined. OBJECTIVE: This study determined the magnitude of change in body weight and composition across sex, race, and age in response to 20 wk of endurance training. DESIGN: Men and women (n = 557) of various ages (16-65 y) and 2 races (black and white) exercised on cycle ergometers 3 d/wk for a total of 60 exercise sessions starting at 55% of maximal oxygen consumption (VO(2)max) for 30 min/session and building to 75% of VO(2)max for 50 min/session, where it was maintained during the last 6 wk. Skinfold-thickness measurements, circumferences, body composition (by hydrostatic weighing), and body fat distribution (by computed tomography scan at L4-L5 and the waist-hip ratio) were determined before and after training. RESULTS: All skinfold-thickness and circumference measures, waist-hip ratio, body mass index, total body mass, fat mass, percentage body fat, and computed tomography scan measures of total, subcutaneous, and visceral abdominal fat decreased with training, whereas total body density and fat-free mass increased. These changes were significant, but small. There were several differences in training response by sex and race, but not by age. CONCLUSIONS: A short-term exercise intervention can induce favorable changes in body composition, but the magnitude of these changes is of limited biological significance. Increasing physical activity likely has a major effect on body composition and fat distribution characteristics only when it is of a greater magnitude and sustained for much longer periods PMID- 10479197 TI - Dietary intake and iron status of Australian vegetarian women. AB - BACKGROUND: Despite the possible overall health benefits of a vegetarian diet, there is concern that some vegetarians and infrequent meat eaters, particularly females, may have inadequate iron status because of low or no heme-iron intakes. OBJECTIVE: The objective was to investigate the nutritional intake and iron status of vegetarian women. DESIGN: The nutritional intakes of 50 free-living vegetarian women aged 18-45 y and 24 age-matched omnivorous control women were assessed by using 12-d weighed dietary records. Iron status was assessed by measuring hemoglobin and serum ferritin concentrations. RESULTS: There was no significant difference between mean (+/-SD) daily iron intakes of vegetarians and omnivores (10.7 +/- 4.4 and 9.9 +/- 2.9 mg, respectively), although heme-iron intakes were low in the vegetarians. Vegetarians had significantly lower intakes of protein (P < 0.01), saturated fat (P < 0.01), and cholesterol (P < 0.001), and significantly higher intakes of dietary fiber (P < 0.001) and vitamin C (P < 0.05). Mean serum ferritin concentrations were significantly lower (P = 0.025) in vegetarians (25.0 +/- 16.2 microg/L) than in omnivores (45.5 +/- 42.5 microg/L). However, similar numbers of vegetarians (18%) and omnivores (13%) had serum ferritin concentrations <12 microg/L, which is a value often used as an indicator of low iron stores. Hemoglobin concentrations were not significantly different. CONCLUSION: It is important that both vegetarian and omnivorous women maintain an adequate iron status and follow dietary practices that enhance iron absorption. PMID- 10479198 TI - Screening of folate status with use of dried blood spots on filter paper. AB - BACKGROUND: Dried blood spots (DBS) on filter paper have been a successful and economical matrix for neonatal screening. OBJECTIVE: Our objective was to develop and evaluate an optimized method for DBS folate analysis and to assess DBS folate stability. DESIGN: DBS were eluted from paper by sonication in 5 g ascorbic acid/L containing 0.1% (by vol) Triton X-100 and hemoglobin folate values (HF; as pmol/g) were calculated from DBS eluate folate and hemoglobin concentrations. RESULTS: Over 95% of DBS folate was eluted during a standardized sonication cycle and DBS folate assay reproducibility was acceptable both within (CV: <8%) and between (CV: <9%) runs. HF means (+/-1 SD) from finger-stick DBS and conventional venous methods were 2513 +/- 1144 and 2607 +/- 1195 pmol/g, respectively, in blood samples taken concurrently from 80 donors, and they correlated well (r = 0.97, P < 0.001). HF values and erythrocyte folate measures may be interconverted by using the mean cell hemoglobin concentration. CONCLUSION: The DBS matrix has potential as an inexpensive and practical option for folate screening studies. PMID- 10479199 TI - Response of vitamin K status to different intakes and sources of phylloquinone rich foods: comparison of younger and older adults. AB - BACKGROUND: Phylloquinone, found in dark-green vegetables and certain plant oils, is the primary dietary source of the fat-soluble vitamin K. Limited data suggest that the relative bioavailability of phylloquinone from vegetables is lower than that from a supplement. This finding is relevant to the maintenance of optimal vitamin K status. OBJECTIVE: The objective of this study was to compare, in younger and older adults, the relative bioavailability of phylloquinone from a vegetable with that of a fortified oil. DESIGN: In a crossover design with three 15-d residency periods in a metabolic unit, younger and older men and women (n = 36) consumed a mixed diet containing 100 microg phylloquinone/d. During 2 residency periods, the mixed diet was supplemented for 5 d with either broccoli (377 microg phylloquinone/d; broccoli diet) or phylloquinone-fortified oil (417 microg/d; oil diet). The relative bioavailability of phylloquinone was defined by the difference in plasma phylloquinone, percentage serum undercarboxylated osteocalcin (%ucOC), and urinary gamma-carboxyglutamic acid in response to 5 d of supplementation. RESULTS: For both younger and older adults, plasma phylloquinone concentrations were higher (P < 0.001) and %ucOC values were lower (P = 0.001) after the broccoli and oil diets than after the mixed diet only. Overall, the response to broccoli supplementation was not significantly different from the response to the fortified oil in either age group. Urinary gamma-carboxyglutamic acid did not change in response to supplementation. CONCLUSIONS: There was no significant difference in the relative bioavailability of phylloquinone, as evidenced by the lack of a significant difference in plasma phylloquinone and %ucOC between the 2 groups after either the broccoli or oil diets for younger and older adults. PMID- 10479200 TI - Prevalence of vitamin K deficiency in cystic fibrosis. AB - BACKGROUND: Patients with cystic fibrosis (CF) are at risk of developing vitamin K deficiency because of pancreatic insufficiency, hepatobiliary disease, or both. OBJECTIVE: Our objective was to determine the prevalence of vitamin K deficiency in unsupplemented patients with CF and to identify risk factors that might be associated with the deficiency. DESIGN: Ninety-eight patients with CF-83 who were pancreatic insufficient (age: 15.2 +/- 10.7 y; range: 0.6-45.8 y), 15 who were pancreatic sufficient (age: 26.2 +/- 11.6 y; range: 6.5-45.3 y), and 62 healthy individuals (age: 16.2 +/- 12. 8 y; range: 1-45 y)-were studied prospectively. None had taken vitamin K supplements. Eight pancreatic-insufficient patients had advanced CF-associated liver disease. Plasma prothrombin in vitamin K absence (PIVKA-II) was measured by immunoassay. All control subjects had PIVKA-II concentrations <3 microg/L. RESULTS: Seventy-eight percent of pancreatic insufficient patients had PIVKA-II concentrations >/=3 microg/L (22.8 +/- 35.7 microg/L). All patients with CF-associated liver disease had abnormal PIVKA-II concentrations. The mean PIVKA-II concentration of pancreatic-insufficient patients with liver disease was greater than that of those without liver disease (46.6 +/- 65.3 compared with 15. 3 +/- 26.1 microg/L; P < 0.05). Five pancreatic sufficient patients had mildly elevated PIVKA-II concentrations. Six (7%) pancreatic insufficient patients (3 with CF-associated liver disease) had mildly prolonged prothrombin time but no clinical bleeding. There was no correlation between PIVKA-II concentrations and severity of fat malabsorption or antibiotic use. CONCLUSIONS: Vitamin K deficiency is common in unsupplemented patients with CF and pancreatic insufficiency and routine supplementation should be considered in all of these patients. PMID- 10479201 TI - trans Fatty acids in human milk are inversely associated with concentrations of essential all-cis n-6 and n-3 fatty acids and determine trans, but not n-6 and n 3, fatty acids in plasma lipids of breast-fed infants. AB - BACKGROUND: Human milk fatty acids vary with maternal dietary fat composition. Hydrogenated dietary oils with trans fatty acids may displace cis n-6 and n-3 unsaturated fatty acids or have adverse effects on their metabolism. The effects of milk trans, n-6, and n-3 fatty acids in breast-fed infants are unclear, although n-6 and n-3 fatty acids are important in infant growth and development. OBJECTIVE: We sought to determine the relations between trans and cis unsaturated fatty acids in milk and plasma phospholipids and triacylglycerols of breast-fed infants, and to identify the major maternal dietary sources of trans fatty acids. DESIGN: We collected milk from 103 mothers with exclusively breast-fed 2-mo-old infants, blood from 62 infants, and 3-d dietary records from 21 mothers. RESULTS: Mean (+/-SEM) percentages of trans fatty acids were as follows: milk, 7.1 +/- 0.32%; infants' triacylglycerols, 6.5 +/- 0. 33%; and infants' phospholipids, 3.7 +/- 0.16%. Milk trans fatty acids, alpha-linolenic acid (18:3n-3), arachidonic acid (20:4n-6), docosahexaenoic acid (22:6n-3) (P < 0.001), and linoleic acid (18:2n-6) (P = 0.007) were each related to the same fatty acid in infant plasma phospholipids. Milk trans fatty acids were inversely related to milk 18:2n-6 and 18:3n-3, but not to milk or infant plasma 20:4n-6 or 22:6n-3. trans Fatty acids represented 7.7% of maternal total fat intake (2.5% of total energy); the major dietary sources were bakery products and breads (32%), snacks (14%), fast foods (11%), and margarines and shortenings (11%). CONCLUSIONS: There were comparable concentrations of trans fatty acids in the maternal diet, breast milk, and plasma triacylglycerols of breast-fed infants. Prepared foods were the major dietary source of trans fatty acids. PMID- 10479202 TI - A randomized, community-based trial of the effects of improved, centrally processed complementary foods on growth and micronutrient status of Ghanaian infants from 6 to 12 mo of age. AB - BACKGROUND: Koko, a fermented maize porridge used as the primary complementary food in Ghana, has been implicated in the high prevalence of child malnutrition. Weanimix, a cereal-legume blend developed by the United Nations Children's Fund and the Ghanaian government, has been promoted as an alternative. OBJECTIVE: We evaluated the effect of feeding Weanimix and 3 other locally formulated, centrally processed complementary foods on the nutritional status of 208 breast fed infants. DESIGN: Infants were randomly assigned to receive 1 of 4 foods from 6 to 12 mo of age: Weanimix (W), Weanimix plus vitamins and minerals (WM), Weanimix plus fish powder (WF), and koko plus fish powder (KF). Dietary and anthropometric data were collected regularly. Blood was collected at 6 and 12 mo of age to assess iron, zinc, vitamin A, and riboflavin status. Before and after the intervention, cross-sectional data on the anthropometric status of infants not included in the intervention (NI; n = 464) were collected. RESULTS: There were no significant differences between intervention groups in weight or length gain or in hemoglobin, hematocrit, transferrin saturation, plasma zinc, or erythrocyte riboflavin values between 6 and 12 mo of age. From 9 to 12 mo of age, z scores were lower in NI infants than in the combined intervention groups [at 12 mo: -1.71 +/- 0.90 compared with -1.19 +/- 0.93 for weight and -1.27 +/- 1.02 compared with -0.63 +/- 0.84 for length (P < 0.001 for both), respectively]. The percentage of infants with low ferritin values increased significantly between 6 and 12 mo of age in groups W, WF, and KF but not in group WM. Change in plasma retinol between 6 and 12 mo of age was significantly greater in group WM than in the other 3 groups combined (0.14 +/- 0.3 compared with -0.04 +/- 0.3 micromol/L, P = 0. 003). CONCLUSIONS: All 4 foods improved growth relative to the NI group. Infants fed WM had better iron stores and vitamin A status than those fed nonfortified foods. PMID- 10479203 TI - Aging, body composition, and lifestyle: the Fels Longitudinal Study. AB - BACKGROUND: Changes in body composition in men and women occur with age, but these changes are affected by numerous covariate factors. OBJECTIVE: The study examined patterns of change in body composition and determined the effects of long-term patterns of change in physical activity in older men and women and in menopausal status and estrogen use in women. DESIGN: Serial measures of height, weight, body mass index (BMI), total body fat (BF), percentage BF, and fat-free mass (FFM) from underwater weighing of 102 men and 108 women enrolled in the Fels Longitudinal Study were analyzed. Physical activity levels and menopausal status were included as covariates. RESULTS: There were significant age-related decreases in FFM and height and increases in total BF, percentage BF, weight, and BMI. Physical activity was associated with decreases in total BF, percentage BF, weight, and BMI in men and were associated with increases in FFM and decreases in total BF and percentage BF in women. Postmenopausal women had significantly higher total BF and percentage BF than did pre- and perimenopausal women. The longer the time since menopause the greater were the increases in weight, BMI, total BF, and percentage BF; however, estrogen use attenuated these increases. CONCLUSIONS: Low FFM can be improved by increased physical activity. The effects of an intervention program on body composition can be masked if only body weight or BMI is measured. The effects of physical activity were more profound in postmenopausal than in premenopausal women, and estrogen use had beneficial effects on body composition. PMID- 10479205 TI - Genetically unknown foods or thrifty genes? PMID- 10479204 TI - Whole-grain consumption and risk of coronary heart disease: results from the Nurses' Health Study. AB - BACKGROUND: Although current dietary guidelines for Americans recommend increased intake of grain products to prevent coronary heart disease (CHD), epidemiologic data relating whole-grain intake to the risk of CHD are sparse. OBJECTIVE: Our objective was to evaluate whether high whole-grain intake reduces risk of CHD in women. DESIGN: In 1984, 75521 women aged 38-63 y with no previous history of cardiovascular disease or diabetes completed a detailed, semiquantitative food frequency questionnaire (SFFQ) and were followed for 10 y, completing SFFQs in 1986 and 1990. We used pooled logistic regression with 2-y intervals to model the incidence of CHD in relation to the cumulative average diet from all 3 cycles of SFFQs. RESULTS: During 729472 person-years of follow-up, we documented 761 cases of CHD (208 of fatal CHD and 553 of nonfatal myocardial infarction). After adjustment for age and smoking, increased whole-grain intake was associated with decreased risk of CHD. For increasing quintiles of intake, the corresponding relative risks (RRs) were 1.0 (reference), 0.86, 0.82, 0.72, and 0.67 (95% CI comparing 2 extreme quintiles: 0.54, 0.84; P for trend < 0.001). After additional adjustment for body mass index, postmenopausal hormone use, alcohol intake, multivitamin use, vitamin E supplement use, aspirin use, physical activity, and types of fat intake, these RRs were 1.0, 0.92, 0.93, 0.83, and 0.75 (95% CI: 0.59, 0.95; P for trend = 0.01). The inverse relation between whole-grain intake and CHD risk was even stronger in the subgroup of never smokers (RR = 0. 49 for extreme quintiles; 95% CI: 0.30, 0.79; P for trend = 0.003). The lower risk associated with higher whole-grain intake was not fully explained by its contribution to intakes of dietary fiber, folate, vitamin B-6, and vitamin E. CONCLUSIONS: Increased intake of whole grains may protect against CHD. PMID- 10479207 TI - Validation of dietary patterns assessed with a food-frequency questionnaire. PMID- 10479209 TI - Very-low-fat diets do not necessarily promote small, dense LDL particles. PMID- 10479211 TI - Preface PMID- 10479212 TI - Appreciation PMID- 10479213 TI - Recognition PMID- 10479214 TI - Dedication UD register, 1920-1997 PMID- 10479215 TI - Convergence of philosophy and science: the third international congress on vegetarian nutrition. AB - Populations of vegetarians living in affluent countries appear to enjoy unusually good health, characterized by low rates of cancer, cardiovascular disease, and total mortality. These important observations have fueled much research and have raised 3 general questions about vegetarians in relation to nonvegetarians: Are these observations the result of better nondietary lifestyle factors, such as lower prevalences of smoking and higher levels of physical activity?; Are they the result of lower intakes of harmful dietary components, in particular meat?; and Are they the result of higher intakes of beneficial dietary components that tend to replace meat in the diet? Current evidence suggests that the answer to all 3 questions is "Yes." Low smoking rates contribute importantly to the low rates of cardiovascular disease and many cancers, probably including colon cancer, in Seventh-day Adventists and other vegetarian populations. Also, avoidance of red meat is likely to account in part for low rates of cardiovascular disease and colon cancer, but this does not appear to be the primary reason for general good health in these populations. Evidence accumulated in the past decade emphasizes the importance of adequate consumption of beneficial dietary factors-rather than just the avoidance of harmful factors including an abundance of fruit, vegetables, and whole grains and regular consumption of vegetable oils, including those from nuts. Although current knowledge already provides general guidance toward healthy diets, accumulated evidence now strongly indicates that diet has a powerful yet complex effect on health and that further investigation is needed. PMID- 10479216 TI - Legumes and soybeans: overview of their nutritional profiles and health effects. AB - Legumes play an important role in the traditional diets of many regions throughout the world. In contrast in Western countries beans tend to play only a minor dietary role despite the fact that they are low in fat and are excellent sources of protein, dietary fiber, and a variety of micronutrients and phytochemicals. Soybeans are unique among the legumes because they are a concentrated source of isoflavones. Isoflavones have weak estrogenic properties and the isoflavone genistein influences signal transduction. Soyfoods and isoflavones have received considerable attention for their potential role in preventing and treating cancer and osteoporosis. The low breast cancer mortality rates in Asian countries and the putative antiestrogenic effects of isoflavones have fueled speculation that soyfood intake reduces breast cancer risk. The available epidemiologic data are limited and only weakly supportive of this hypothesis, however, particularly for postmenopausal breast cancer. The data suggesting that soy or isoflavones may reduce the risk of prostate cancer are more encouraging. The weak estrogenic effects of isoflavones and the similarity in chemical structure between soybean isoflavones and the synthetic isoflavone ipriflavone, which was shown to increase bone mineral density in postmenopausal women, suggest that soy or isoflavones may reduce the risk of osteoporosis. Rodent studies tend to support this hypothesis, as do the limited preliminary data from humans. Given the nutrient profile and phytochemical contribution of beans, nutritionists should make a concerted effort to encourage the public to consume more beans in general and more soyfoods in particular. PMID- 10479217 TI - Cereals, legumes, and chronic disease risk reduction: evidence from epidemiologic studies. AB - There is growing evidence that cereals and legumes play important roles in the prevention of chronic diseases. Early epidemiologic studies of these associations focused on intake of dietary fiber rather than intake of grains or legumes. Generally, these studies indicated an inverse association between dietary fiber intake and risk of coronary artery disease; this observation has been replicated in recent cohort studies. Studies that focused on grain or cereal intake are fewer in number; these tend to support an inverse association between intake of whole grains and coronary artery disease. Studies on the association of dietary fiber with colon and other cancers have generally shown inverse relations, but whether these relations are attributable to cereals, other fiber sources, or other factors is less clear. Although legumes have been shown to lower blood cholesterol concentrations, epidemiologic studies are few and inconclusive regarding the association of legumes with risk of coronary artery disease. It has been hypothesized that legumes, in particular soybeans, reduce the risk of some cancers, but epidemiologic studies are equivocal in this regard. Overall, there is substantial epidemiologic evidence that dietary fiber and whole grains are associated with decreased risk of coronary artery disease and some cancers, whereas the role of legumes in these diseases appears promising but as yet inconclusive. PMID- 10479218 TI - Plausible mechanisms for the protectiveness of whole grains. AB - Dietary guidelines recommend the consumption of whole grains to prevent chronic diseases. Epidemiologic studies support the theory that whole grains are protective against cancer, especially gastrointestinal cancers such as gastric and colon can-cer, and cardiovascular disease. Components in whole grains that may be protective include compounds that affect the gut environment, such as dietary fiber, resistant starch, and oligosaccharides. Whole grains are also rich in compounds that function as antioxidants, such as trace minerals and phenolic compounds, and phytoestrogens, with potential hormonal effects. Other potential mechanisms whereby whole grains may protect against disease include binding of carcinogens and modulation of the glycemic response. Clearly, the range of protective substances in whole grains is impressive and advice to consume additional whole grains is justified. Further study is needed regarding the mechanisms behind this protection so that the most potent protective components of whole grains will be maintained when developing whole grains into acceptable food products for the public. PMID- 10479219 TI - Cardiovascular and renal benefits of dry bean and soybean intake. AB - Dry beans and soybeans are nutrient-dense, fiber-rich, and are high-quality sources of protein. Protective and therapeutic effects of both dry bean and soybean intake have been documented. Studies show that dry bean intake has the potential to decrease serum cholesterol concentrations, improve many aspects of the diabetic state, and provide metabolic benefits that aid in weight control. Soybeans are a unique source of the isoflavones genistein and diadzein, which have numerous biological functions. Soybeans and soyfoods potentially have multifaceted health-promoting effects, including cholesterol reduction, improved vascular health, preserved bone mineral density, and reduction of menopausal symptoms. Soy appears to have salutary effects on renal function, although these effects are not well understood. Whereas populations consuming high intakes of soy have lower prevalences of certain cancers, definitive experimental data are insufficient to clarify a protective role of soy. The availability of legume products and resources is increasing, incorporating dry beans and soyfoods into the diet can be practical and enjoyable. With the shift toward a more plant-based diet, dry beans and soy will be potent tools in the treatment and prevention of chronic disease. PMID- 10479220 TI - Health effects of vegetables and fruit: assessing mechanisms of action in human experimental studies. AB - Epidemiologic data support the association between high intake of vegetables and fruits and low risk of chronic disease. There are several biologically plausible reasons why consumption of vegetables and fruit might slow or prevent the onset of chronic diseases. Vegetables and fruit are rich sources of a variety of nutrients, including vitamins, trace minerals, and dietary fiber, and many other classes of biologically active compounds. These phytochemicals can have complementary and overlapping mechanisms of action, including modulation of detoxification enzymes, stimulation of the immune system, reduction of platelet aggregation, modulation of cholesterol synthesis and hormone metabolism, reduction of blood pressure, and antioxidant, antibacterial, and antiviral effects. Although these effects have been examined primarily in animal and cell culture models, experimental dietary studies in humans have also shown the capacity of vegetables and fruit and their constituents to modulate some of these potential disease-preventive mechanisms. The human studies have relied on intermediate endpoints related to disease risk. Design methodologies used include multiple-arm trials, randomized crossover studies, and more compromised designs such as nonrandomized crossovers and pre- and posttreatment analyses. Length of treatment ranged from a single dose to years depending on the mechanism of interest. Stringency of dietary control varied from addition of supplements to a habitual diet to provision of all food for the duration of a treatment. Rigorously conducted experimental dietary studies in humans are an important link between population- and laboratory-based research. PMID- 10479221 TI - Health-promoting properties of common herbs. AB - Herbs have been used as food and for medicinal purposes for centuries. Research interest has focused on various herbs that possess hypolipidemic, antiplatelet, antitumor, or immune-stimulating properties that may be useful adjuncts in helping reduce the risk of cardiovascular disease and cancer. In different herbs, a wide variety of active phytochemicals, including the flavonoids, terpenoids, lignans, sulfides, polyphenolics, carotenoids, coumarins, saponins, plant sterols, curcumins, and phthalides have been identified. Several of these phytochemicals either inhibit nitrosation or the formation of DNA adducts or stimulate the activity of protective enzymes such as the Phase II enzyme glutathione transferase (EC 2.5.1.18). Research has centered around the biochemical activity of the Allium sp. and the Labiatae, Umbelliferae, and Zingiberaceae families, as well as flaxseed, licorice root, and green tea. Many of these herbs contain potent antioxidant compounds that provide significant protection against chronic diseases. These compounds may protect LDL cholesterol from oxidation, inhibit cyclooxygenase and lipoxygenase enzymes, inhibit lipid peroxidation, or have antiviral or antitumor activity. The volatile essential oils of commonly used culinary herbs, spices, and herbal teas inhibit mevalonate synthesis and thereby suppress cholesterol synthesis and tumor growth. PMID- 10479222 TI - Nut consumption, vegetarian diets, ischemic heart disease risk, and all-cause mortality: evidence from epidemiologic studies. AB - Perhaps one of the most unexpected and novel findings in nutritional epidemiology in the past 5 y has been that nut consumption seems to protect against ischemic heart disease (IHD). Frequency and quantity of nut consumption have been documented to be higher in vegetarian than in nonvegetarian populations. Nuts also constitute an important part of other plant-based diets, such as Mediterranean and Asian diets. In a large, prospective epidemiologic study of Seventh-day Adventists in California, we found that frequency of nut consumption had a substantial and highly significant inverse association with risk of myocardial infarction and death from IHD. The Iowa Women's Health Study also documented an association between nut consumption and decreased risk of IHD. The protective effect of nuts on IHD has been found in men and women and in the elderly. Importantly, nuts have similar associations in both vegetarians and nonvegetarians. The protective effect of nut consumption on IHD is not offset by increased mortality from other causes. Moreover, frequency of nut consumption has been found to be inversely related to all-cause mortality in several population groups such as whites, blacks, and the elderly. Thus, nut consumption may not only offer protection against IHD, but also increase longevity. PMID- 10479223 TI - Nuts and their bioactive constituents: effects on serum lipids and other factors that affect disease risk. AB - Because nuts have favorable fatty acid and nutrient profiles, there is growing interest in evaluating their role in a heart-healthy diet. Nuts are low in saturated fatty acids and high in monounsaturated and polyunsaturated fatty acids. In addition, emerging evidence indicates that there are other bioactive molecules in nuts that elicit cardioprotective effects. These include plant protein, dietary fiber, micronutrients such as copper and magnesium, plant sterols, and phytochemicals. Few feeding studies have been conducted that have incorporated different nuts into the test diets to determine the effects on plasma lipids and lipoproteins. The total- and lipoprotein-cholesterol responses to these diets are summarized in this article. In addition, the actual cholesterol response was compared with the predicted response derived from the most current predictive equations for blood cholesterol. Results from this comparison showed that when subjects consumed test diets including nuts, there was an approximately 25% greater cholesterol-lowering response than that predicted by the equations. These results suggest that there are non-fatty acid constituents in nuts that have additional cholesterol-lowering effects. Further studies are needed to identify these constituents and establish their relative cholesterol-lowering potency. PMID- 10479224 TI - The role of dietary fats in plant-based diets. AB - In the United States, the notion that low-fat, high-carbohydrate diets are essential for health has grown into an obsession, driven largely by an effort to reduce heart disease and, more recently, certain types of cancer. We know that saturated fatty acids are more closely associated with risk factors for heart disease than are unsaturated fatty acids. Many people believe that plant-based diets are healthy because they are low in fat. However, plant-based diets are not necessarily low-fat. In true plant-based diets, unsaturated fatty acids predominate, whereas saturated fatty acids come largely from animal sources such as dairy products and eggs. Plant-based diets include foods that contain fats, such as nuts and seeds and oils from grains and seeds. The fats in these foods are not associated with increased risk for heart disease. In addition, for people with insulin resistance, higher-fat diets protect against the heart disease risk factors of low HDL-cholesterol concentration, hypertriglyceridemia, hyperglycemia, and hyperinsulinemia. Because humans can synthesize fat from dietary carbohydrate, and because our adipose stores and circulating fatty acids reflect dietary intake, scientists understand the relations between the amounts and types of dietary fats and the types of fats found in body fat depots. Consuming dietary fats that are not associated with increased risk of disease can be a part of a healthful diet. PMID- 10479225 TI - Mortality in vegetarians and nonvegetarians: detailed findings from a collaborative analysis of 5 prospective studies. AB - We combined data from 5 prospective studies to compare the death rates from common diseases of vegetarians with those of nonvegetarians with similar lifestyles. A summary of these results was reported previously; we report here more details of the findings. Data for 76172 men and women were available. Vegetarians were those who did not eat any meat or fish (n = 27808). Death rate ratios at ages 16-89 y were calculated by Poisson regression and all results were adjusted for age, sex, and smoking status. A random-effects model was used to calculate pooled estimates of effect for all studies combined. There were 8330 deaths after a mean of 10.6 y of follow-up. Mortality from ischemic heart disease was 24% lower in vegetarians than in nonvegetarians (death rate ratio: 0.76; 95% CI: 0.62, 0.94; P<0.01). The lower mortality from ischemic heart disease among vegetarians was greater at younger ages and was restricted to those who had followed their current diet for >5 y. Further categorization of diets showed that, in comparison with regular meat eaters, mortality from ischemic heart disease was 20% lower in occasional meat eaters, 34% lower in people who ate fish but not meat, 34% lower in lactoovovegetarians, and 26% lower in vegans. There were no significant differences between vegetarians and nonvegetarians in mortality from cerebrovascular disease, stomach cancer, colorectal cancer, lung cancer, breast cancer, prostate cancer, or all other causes combined. PMID- 10479226 TI - The Oxford Vegetarian Study: an overview. AB - The Oxford Vegetarian Study is a prospective study of 6000 vegetarians and 5000 nonvegetarian control subjects recruited in the United Kingdom between 1980 and 1984. Cross-sectional analyses of study data showed that vegans had lower total- and LDL-cholesterol concentrations than did meat eaters; vegetarians and fish eaters had intermediate and similar values. Meat and cheese consumption were positively associated, and dietary fiber intake was inversely associated, with total-cholesterol concentration in both men and women. After 12 y of follow-up, all-cause mortality in the whole cohort was roughly half that in the population of England and Wales (standardized mortality ratio, 0.46; 95% CI, 0.42, 0.51). After adjusting for smoking, body mass index, and social class, death rates were lower in non-meat-eaters than in meat eaters for each of the mortality endpoints studied [relative risks and 95% CIs: 0.80 (0. 65, 0.99) for all causes of death, 0.72 (0.47, 1.10) for ischemic heart disease, and 0.61 (0.44, 0.84) for all malignant neoplasms]. Mortality from ischemic heart disease was also positively associated with estimated intakes of total animal fat, saturated animal fat, and dietary cholesterol. Other analyses showed that non-meat-eaters had only half the risk of meat eaters of requiring an emergency appendectomy, and that vegans in Britain may be at risk for iodine deficiency. Thus, the health of vegetarians in this study is generally good and compares favorably with that of the nonvegetarian control subjects. Larger studies are needed to examine rates of specific cancers and other diseases among vegetarians. PMID- 10479227 TI - Associations between diet and cancer, ischemic heart disease, and all-cause mortality in non-Hispanic white California Seventh-day Adventists. AB - Results associating diet with chronic disease in a cohort of 34192 California Seventh-day Adventists are summarized. Most Seventh-day Adventists do not smoke cigarettes or drink alcohol, and there is a wide range of dietary exposures within the population. About 50% of those studied ate meat products <1 time/wk or not at all, and vegetarians consumed more tomatoes, legumes, nuts, and fruit, but less coffee, doughnuts, and eggs than did nonvegetarians. Multivariate analyses showed significant associations between beef consumption and fatal ischemic heart disease (IHD) in men [relative risk (RR) = 2.31 for subjects who ate beef > or =3 times/wk compared with vegetarians], significant protective associations between nut consumption and fatal and nonfatal IHD in both sexes (RR approximately 0.5 for subjects who ate nuts > or =5 times/wk compared with those who ate nuts <1 time/wk), and reduced risk of IHD in subjects preferring whole-grain to white bread. The lifetime risk of IHD was reduced by approximately 31% in those who consumed nuts frequently and by 37% in male vegetarians compared with nonvegetarians. Cancers of the colon and prostate were significantly more likely in nonvegetarians (RR of 1.88 and 1.54, respectively), and frequent beef consumers also had higher risk of bladder cancer. Intake of legumes was negatively associated with risk of colon cancer in nonvegetarians and risk of pancreatic cancer. Higher consumption of all fruit or dried fruit was associated with lower risks of lung, prostate, and pancreatic cancers. Cross-sectional data suggest vegetarian Seventh-day Adventists have lower risks of diabetes mellitus, hypertension, and arthritis than nonvegetarians. Thus, among Seventh-day Adventists, vegetarians are healthier than nonvegetarians but this cannot be ascribed only to the absence of meat. PMID- 10479228 TI - Plant-based diets and bone health: nutritional implications. AB - Hip fracture incidence rates are predicted to increase dramatically in the first half of the 21st century, especially in Asia, Latin America, Africa, and the Middle East. These increased rates will result primarily from the effects of public health efforts to improve nutrition and infectious-disease control, both of which contribute to improved longevity of populations. An example of a rapid increase in hip fracture incidence rates has been reported in Hong Kong. Findings of studies there suggest that environmental changes, ie, westernization, urbanization, or both, are strongly related with declines in bone mineral density and increases in fractures. Hip fracture incidence rates in Western nations are typically increasing at much more modest rates than those in Hong Kong and other Asian nations. Epidemiologic investigations have identified multiple risk factors, including exposures earlier in life to adverse factors that are considered to contribute to the development of osteoporosis in both Western and Asian nations. The major risk factors are inadequate nutrition, limited physical activity, and low lifetime estrogen exposure. A dietary shift toward a more plant based diet in Western nations may be beneficial to bone health, but is not likely to counter the adverse effects of limited physical activity and low estrogen exposure. PMID- 10479229 TI - Choices for achieving adequate dietary calcium with a vegetarian diet. AB - To achieve adequate dietary calcium intake, several choices are available that accommodate a variety of lifestyles and tastes. Liberal consumption of dairy products in the diet is the approach of most Americans. Some plants provide absorbable calcium, but the quantity of vegetables required to reach sufficient calcium intake make an exclusively plant-based diet impractical for most individuals unless fortified foods or supplements are included. Also, dietary constituents that decrease calcium retention, such as salt, protein, and caffeine, can be high in the vegetarian diet. Although it is possible to obtain calcium balance from a plant-based diet in a Western lifestyle, it may be more convenient to achieve calcium balance by increasing calcium consumption than by limiting other dietary factors. PMID- 10479230 TI - Vegetarianism and menstrual cycle disturbances: is there an association? AB - The question of whether menstrual disturbances are more common in vegetarian than in nonvegetarian women is complex. Disturbances of the cycle may be clinical (ie, amenorrhea or oligomenorrhea) or subclinical (i.e., normal-length cycles with anovulation or a short or defective luteal phase). Detection of the latter requires that the menstrual cycle be monitored, but may help prevent recruitment bias in studies comparing vegetarians with nonvegetarians because vegetarians with menstrual disturbances may be more likely to volunteer for a study on menstrual disturbances and vegetarianism. Three general mechanisms that could contribute to menstrual disturbances that may differ between vegetarians and nonvegetarians include energy imbalances associated with body-weight disturbances or exercise, psychosocial and cognitive factors, and dietary components. Evidence for each of these mechanisms is reviewed and studies comparing menstrual function between vegetarians and nonvegetarians are described in this article. Although results from several cross-sectional studies suggest that clinical menstrual disturbances may be more common in vegetarians, a prospective study that controlled for many potential confounders found that subclinical disturbances were less common in weight-stable, healthy vegetarian women. Because the sample studied may not be representative of all vegetarian women, however, these results cannot be generalized. Population studies are needed to draw definitive conclusions. PMID- 10479231 TI - Essential fatty acid requirements of vegetarians in pregnancy, lactation, and infancy. AB - Long-chain polyunsaturated fatty acids (LCPUFAs) derived from linoleic (18:2n-6) and alpha-linolenic (18:3n-3) acids are required for the normal development of the retina and central nervous system, but the extent to which they can be synthesized from the parent fatty acids is debated. Consuming LCPUFAs markedly increases their proportions in tissue lipids compared with their parent fatty acids. Thus, it has been argued that LCPUFAs must be supplied in the diet. LCPUFAs are generally absent from plant foods, thus it is important find out how essential fatty acid requirements are met by vegetarians. A developing fetus obtains LCPUFAs via selective uptake from its mother's plasma and LCPUFAs are present in the breast milk of vegetarians. There is no evidence that the capacity to synthesize LCPUFAs is limited in vegetarians. However, there are greater proportions of n-6 LCPUFAs and lower proportions of n-3 LCPUFAs in vegetarians compared with omnivores. This difference is probably a consequence of the selection of foods by vegetarians with high amounts of linoleic acid. Although lower concentrations of docosahexaenoic acid (22:6n-3; DHA) have been observed in blood and artery phospholipids of infants of vegetarians, it is uncertain whether their brain lipids contain lower proportions of DHA than do those of infants of omnivores. On the basis of experiments in primates that showed altered visual function with a high ratio of linoleic acid to alpha-linolenic acid, it would be prudent to recommend diets with a ratio between 4:1 and 10:1 in vegetarians and that excessive intakes of linoleic acid be avoided. PMID- 10479232 TI - Essential fatty acids in health and chronic disease. AB - Human beings evolved consuming a diet that contained about equal amounts of n-3 and n-6 essential fatty acids. Over the past 100-150 y there has been an enormous increase in the consumption of n-6 fatty acids due to the increased intake of vegetable oils from corn, sunflower seeds, safflower seeds, cottonseed, and soybeans. Today, in Western diets, the ratio of n-6 to n-3 fatty acids ranges from approximately 20-30:1 instead of the traditional range of 1-2:1. Studies indicate that a high intake of n-6 fatty acids shifts the physiologic state to one that is prothrombotic and proaggregatory, characterized by increases in blood viscosity, vasospasm, and vasoconstriction and decreases in bleeding time. n-3 Fatty acids, however, have antiinflammatory, antithrombotic, antiarrhythmic, hypolipidemic, and vasodilatory properties. These beneficial effects of n-3 fatty acids have been shown in the secondary prevention of coronary heart disease, hypertension, type 2 diabetes, and, in some patients with renal disease, rheumatoid arthritis, ulcerative colitis, Crohn disease, and chronic obstructive pulmonary disease. Most of the studies were carried out with fish oils [eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)]. However, alpha linolenic acid, found in green leafy vegetables, flaxseed, rapeseed, and walnuts, desaturates and elongates in the human body to EPA and DHA and by itself may have beneficial effects in health and in the control of chronic diseases. PMID- 10479233 TI - Physical fitness and vegetarian diets: is there a relation? AB - The available evidence supports neither a beneficial nor a detrimental effect of a vegetarian diet on physical performance capacity, especially when carbohydrate intake is controlled for. Concerns have been raised that an emphasis on plant foods to enhance carbohydrate intake and optimize body glycogen stores may lead to increases in dietary fiber and phytic acid intake to concentrations that reduce the bioavailability of several nutrients, including zinc, iron, and some other trace minerals. There is no convincing evidence, however, that vegetarian athletes suffer impaired nutrient status from the interactive effect of their heavy exertion and plant-food based dietary practices to the extent that performance, health, or both are impaired. Although there has been some concern about protein intake for vegetarian athletes, data indicate that all essential and nonessential amino acids can be supplied by plant food sources alone as long as a variety of foods is consumed and the energy intake is adequate. There has been some concern that vegetarian female athletes are at increased risk for oligoamenorrhea, but evidence suggests that low energy intake, not dietary quality, is the major cause. In conclusion, a vegetarian diet per se is not associated with improved aerobic endurance performance. Although some concerns have been raised about the nutrient status of vegetarian athletes, a varied and well-planned vegetarian diet is compatible with successful athletic endeavor. PMID- 10479234 TI - Cyanocobalamin (vitamin B-12) status in Seventh-day Adventist ministers in Australia. AB - As part of the Adventist Ministers' Health Study, a series of cross-sectional surveys conducted in 1992, 1994, and 1997, the serum vitamin B-12 status of 340 Australian Seventh-day Adventist ministers was assessed in 1997. The ministers in the study participated voluntarily. Of this group, 245 were either lactoovovegetarians or vegans who were not taking vitamin B-12 supplements. Their mean vitamin B-12 concentration was 199 pmol/L (range: 58-538 pmol/L), 53% of whom had values below the reference range for the method used (171-850 pmol/L) and 73% of whom had values <221 pmol/L, the lower limit recommended by Herbert. Dual-isotope Schillings test results in 36 lactoovovegetarians with abnormally low vitamin B-12 concentrations indicated that dietary deficiency was the cause in 70% of cases. Data from the dietary questionnaires supported dietary deficiency as the cause of low serum vitamin B-12 in this population of lactoovovegetarians and vegans, 56 (23%) of whom consumed sufficient servings of vitamin B-12-containing foods to obtain the minimum daily maintenance allowance of the vitamin (1 microg). PMID- 10479235 TI - Growth, development, and physical fitness of Flemish vegetarian children, adolescents, and young adults. AB - This study was designed to assess average daily dietary intakes of energy in 82 vegetarian children (group A: 6- 9-y-old girls and 6-11-y-old boys), adolescents (group B: 10- 15-y-old girls and 12-17-y-old boys), and young adults (group C: 16 30-y-old females and 18-30-y-old males) and included determination of height and weight; triceps, suprailiac, and calf skinfold thicknesses; puberty ratings; and physical fitness. Dietary energy intake was lower than recommended values in all 3 groups. Height and weight did not differ significantly from the reference data except in group B, which had significantly lower heights and weights and lower body mass indexes (P<0.05). Triceps and suprailiac skinfold thicknesses were lower in all age groups, whereas the calf skinfold thickness was only significantly lower in the 10-15-y-old girls (P<0.05). The vegetarian children were as physically fit as the reference group. The vegetarian adolescent boys and girls and the young adults scored significantly lower on the standing long jump and 30-s sit-up (P<0.05). The vegetarian subjects of groups B and C recovered significantly faster from the step test (P<0.05). Puberty ratings plotted on percentile graphs showed that all vegetarian subjects, except for 1 girl, were within the normal developmental range. We conclude that, within the limits of this study, vegetarian subjects have lower relative body weights and skinfold thicknesses in adolescence than do nonvegetarians. They scored lower on the strength tests and better on the cardiorespiratory test when compared with reference values. The growth and maturation status of the vegetarian population were within the normal range. PMID- 10479236 TI - Dietary intake and biochemical, hematologic, and immune status of vegans compared with nonvegetarians. AB - Dietary and nutritional status of individuals habitually consuming a vegan diet was evaluated by biochemical, hematologic, and immunologic measures in comparison with a nonvegetarian group. On the basis of 4-d dietary records, the intake of female and male vegans tended to be lower in fat, saturated fat, monounsaturated fat, and cholesterol and higher in dietary fiber than that of vegetarians. With computed food and supplement intakes, vegan diets provided significantly higher amounts of ascorbate, folate, magnesium, copper, and manganese in both female and male participants. The body mass index (BMI; in kg/m(2)) of the vegans was significantly lower than that of the nonvegetarians and 9 of the 25 vegans had a BMI <19. Serum ferritin concentrations were significantly lower in vegan men but iron and zinc status did not differ between the sexes. Mean serum vitamin B-12 and methylmalonic acid concentrations did not differ; however, 10 of the 25 vegans showed a vitamin B-12 deficit manifested by macrocytosis, circulating vitamin B-12 concentrations <150 pmol/L, or serum methylmalonic acid >376 nmol/L. Vegans had significantly lower leukocyte, lymphocyte, and platelet counts and lower concentrations of complement factor 3 and blood urea nitrogen but higher serum albumin concentrations. Vegans did not differ from nonvegetarians in functional immunocompetence assessed as mitogen stimulation or natural killer cell cytotoxic activity. PMID- 10479237 TI - Rheumatoid arthritis treated with vegetarian diets. AB - The notion that dietary factors may influence rheumatoid arthritis (RA) has been a part of the folklore of the disease, but scientific support for this has been sparse. In a controlled, single-blind trial we tested the effect of fasting for 7 10 d, then consuming an individually adjusted, gluten-free, vegan diet for 3.5 mo, and then consuming an individually adjusted lactovegetarian diet for 9 mo on patients with RA. For all clinical variables and most laboratory variables measured, the 27 patients in the fasting and vegetarian diet groups improved significantly compared with the 26 patients in the control group who followed their usual omnivorous diet throughout the study period. One year after the patients completed the trial, they were reexamined. Compared with baseline, the improvements measured were significantly greater in the vegetarians who previously benefited from the diet (diet responders) than in diet nonresponders and omnivores. The beneficial effect could not be explained by patients' psychologic characteristics, antibody activity against food antigens, or changes in concentrations of prostaglandin and leukotriene precursors. However, the fecal flora differed significantly between samples collected at time points at which there was substantial clinical improvement and time points at which there were no or only minor improvements. In summary, the results show that some patients with RA can benefit from a fasting period followed by a vegetarian diet. Thus, dietary treatment may be a valuable adjunct to the ordinary therapeutic armamentarium for RA. PMID- 10479238 TI - Publication trends of vegetarian nutrition articles in biomedical literature, 1966-1995. AB - We documented publication trends of vegetarian nutrition articles in biomedical literature between 1966 and 1995 using the National Institutes of Health MEDLINE bibliographic database. The publication rate of vegetarian articles increased steadily during the 3 decades, from an average of <10/y in the late 1960s to 76/y in the early 1990s. After adjusting for the total number of articles indexed in MEDLINE annually, we found that publication of vegetarian nutrition articles increased dramatically, by 4-fold, during the 1970s and reached an oscillating plateau during the 1980s. In the early 1990s, the proportion of vegetarian nutrition articles 8 articles per 1000 vegetarian nutrition articles and approximately 20 per 100000 articles indexed by MEDLINE. Non-nutrition journals have progressively published a larger share of all vegetarian articles in the biomedical literature during the period studied. The nature and study design of published vegetarian research has changed over the years as well. The proportion of original research and review articles increased whereas case series and letters to the editor decreased. Reports of epidemiologic studies of vegetarians with longitudinal designs have superseded cross-sectional designs in number and proportion. In 40% of all publications, preventive and therapeutic applications of vegetarian diets constituted the major themes of vegetarian articles in the decade of 1986-1995. However, 20 y earlier the main focus was on the nutritional adequacy of vegetarian diets. The progressive change in the themes of vegetarian nutrition publications is interpreted as a shift in the role of vegetarian diets in human nutrition. PMID- 10479239 TI - Complex systems model of dietary choice with implications for improving diets and promoting vegetarianism. AB - An important step toward improving nutrition and promoting vegetarianism in the general population is to understand how consumers make dietary choices. Researchers from many clinical and social sciences are interested in dietary choice but have not combined their research into a comprehensive model to explain consumer actions. No one model has offered a good explanation for the fact that, although many people successfully change their diet significantly (often toward health-improving, plant-based diets) and are happy with the change, the public and health professionals often perceive dietary change as being difficult and unlikely to succeed. I have termed these observations "the paradox of dietary change." The present computer model uses the emerging science of complex systems analysis, which offers an intuitive method for studying evidence about dietary choice from many fields, including public health, clinical science, economics, sociology, marketing, and genetics, and for combining individual choice with social interaction. The results suggest an explanation for the paradox and methods for helping society shift toward healthier and more plant-based diets. In particular, they suggest how and why major changes might be easier to make than incremental ones, and why this makes dietary change seem more difficult to consumers than it actually is. PMID- 10479240 TI - Vegetarian food guide pyramid: a conceptual framework. AB - The purpose of this article and the accompanying vegetarian food guide pyramid graphic is to provide the conceptual framework for the development of a new and unique food guide. Food guides for vegetarians have tended to be adaptations of guides developed for the general nonvegetarian population instead of being designed to emphasize the healthy components of vegetarian dietary patterns. A subcommittee of the organizers of the Third International Congress on Vegetarian Nutrition began a process that led to the development of a pyramid-shaped graphic illustration and a supporting document, both of which were introduced at the congress. The 5 major plant-based food groups (whole grains, legumes, vegetables, fruit, nuts, and seeds) form the trapezoid-shaped lower portion of the pyramid. Optional food groups, which may be avoided by some vegetarians (vegetable oils, dairy, eggs, and sweets), form the smaller, separate, triangle-shaped top portion of the pyramid. The supporting document discusses the concepts that affect vegetarian food guidance and the rationale for selecting the food groups. It is hoped that this framework will provide the impetus for further research and discussion and will lead to the development of a guide that is nutritionally adequate, is conducive to good health, and can be adopted by vegetarians of diverse eating practices. PMID- 10479241 TI - Convergence of plant-rich and plant-only diets. AB - Discussants at the Third International Congress on Vegetarian Nutrition considered the nutritional adequacy, benefits, and health outcomes of plant-only (e.g., vegan and fruitarian), plant-based (e.g., macrobiotic, lactovegetarian, semivegetarian, and meatless), and omnivorous dietary patterns. The increased availability of a variety of plant foods, the advent of nutrient-fortified plant foods, the use of vitamin and mineral supplements, and the widespread dissemination of sound information on dietary patterns mean that convergence between the essential nutrient profiles of plant-only and plant-rich, plant-based diets is possible. Special attention should be paid to nutrition among vulnerable groups by age or physiologic status if they consume diets based solely on plants. Research has shown that both plant-only and plant-based eating patterns have health benefits, most notably in reducing the risk of chronic, degenerative diseases. The panel concluded that evidence for a convergence of scientific opinion on the safety and healthfulness of plant-only diets that are appropriately planned to meet all nutrient requirements compared with plant-based diets is considerable. PMID- 10479243 TI - II nutritional status and life cycle issues PMID- 10479242 TI - I diet and chronic disease PMID- 10479245 TI - IV applied vegetarian nutrition PMID- 10479244 TI - III biochemical and clinical outcomes PMID- 10479246 TI - Surgical template for crown lengthening: A clinical report. PMID- 10479247 TI - Implant overdenture resolution of a Class II skeletal relationship of the edentulous patient: A clinical report. PMID- 10479248 TI - Myeloperoxidase deficiency associated with atypical oral candidiasis: A clinical report. PMID- 10479249 TI - Complications of surgical crown lengthening for a maxillary molar with four roots: A clinical report. PMID- 10479250 TI - Accidental impaction of a unilateral removable partial denture: A clinical report. PMID- 10479251 TI - Diametral and compressive strength of dental core materials. AB - STATEMENT OF PROBLEM: Strength greatly influences the selection of core materials. Many disparate material types are now recommended for use as cores. Cores must withstand forces due to mastication and parafunction for many years. PURPOSE: This study compared the compressive and diametral tensile strengths of 8 core materials of various material classes and formulations (light-cured hybrid composite, autocured titanium containing composite, amalgam, glass ionomer, glass ionomer cermet, resin-modified glass ionomer, and polyurethane). MATERIAL AND METHODS: Materials were manipulated according to manufacturers' instructions for use as cores. Mean compressive and diametral strengths with associated standard errors were calculated for each material (n = 10). Analyses of variance were computed (P <.0001) and multiple comparisons tests discerned many differences among materials. RESULTS: Compressive strengths varied widely from 61.1 MPa for a polyurethane to 250 MPa for a resin composite. Diametral tensile strengths ranged widely from 18.3 MPa for a glass ionomer cermet to 55.1 MPa for a resin composite. Some resin composites had compressive and tensile strengths equal to those of amalgam. CONCLUSION: Light-cured hybrid resin composites were stronger than autocured titanium containing composites. The strengths of glass ionomer based materials and of a polyurethane material were considerably lower than for resin composites or amalgam. PMID- 10479252 TI - Pilot study of the effects of three bonding systems on the torsional resistance of a titanium-reinforced composite core. AB - STATEMENT OF PROBLEM: There are no studies analyzing the effects of a bonding agent on torsional resistance for a titanium-reinforced core material bonded with a multistep bonding system. PURPOSE: This pilot study investigated the effects of multistep bonding agents on torsional resistance of a titanium-reinforced composite core material (Ti-Core core material). MATERIAL AND METHODS: A total of 40 recently extracted human molars with their crowns removed were placed into acrylic resin blocks and divided into 4 groups of 10 specimens. Only multistep bonding systems were used (AllBond-2, ScotchBond Multi- PURPOSE: Tenure A&B, and the control without bonding agent). Specimens were prepared according to manufacturer's instructions. The control group was etched with a 37.5% phosphate acid. All cores were fabricated with Ti-Core titanium core material with No. 1 hard copper bands as the matrix. Ti-Core core material was applied and allowed to cure for 1 hour, then the cores were placed in 100% humidity (water) for 1 week. Test specimens were placed in a special jig and a clockwise torsional force applied. Torsional force was measured on a Lebow 1102-200 torque transducer and recorded on a Hewlett Packard 7015B X-Y recorder in inch x ounce then converted into Newton-meter. One-way analysis of variance (ANOVA) was used to test for statistical significance (P <.05). RESULTS: Torsional values (Newton-meter) were AllBond-2, 0.1980, ScotchBond Multi- PURPOSE: 0.1890, Tenure A&B, 0.2142; the control group recorded a value of zero. Statistical analysis revealed that bonding systems did not differ from one another. CONCLUSION: Use of a bonding agent increased the core's resistance to torsional forces. Lack of a bonding agent dramatically reduced the resistance to torque. PMID- 10479253 TI - Bonding porcelain laminate veneer provisional restorations: An experimental study. AB - STATEMENT OF PROBLEM: Minimal tooth preparation is required for porcelain laminate veneers, but an interim restoration is anticipated by patients to protect their teeth against thermal insult, chemical irritation, and to provide esthetics. Cement remaining after the removal of the provisional restoration can impair the etching quality of the tooth surface and the fit and final bonding of the porcelain laminate veneer. PURPOSE: This in vitro study inspected the tooth surface for remaining debris from cement after removal of a provisional restoration. MATERIAL AND METHODS: Seventy-two extracted natural anterior teeth were prepared for porcelain laminate veneers. For half of the teeth, the smear layer was removed before luting provisional restorations. Veneer provisional restorations were fabricated and luted to teeth with 3 bonding methods: temporary eugenol-free cement, spot etching combined with dual-curing luting cement, and polyurethane adhesive combined with dual-curing luting cement. After removal of provisional restorations 1 week later, the tooth surface was examined for residual luting material with an SEM. RESULTS: Traces of cement debris were found on provisionally prepared teeth for all 3 material methods. CONCLUSION: The use of polyurethane adhesive combined with the dual-curing cement revealed significantly less teeth with debris than the other methods. This difference was less distinct when the smear layer was removed. PMID- 10479254 TI - Comparision of diametral tensile strength of microwave and oven-dried investment materials. AB - STATEMENT OF PROBLEM: Microwave drying technique for investment materials is believed to be timesaving by accelerating the elimination of free water content. PURPOSE: This study compared the diametral tensile strength of 4 investment materials used in removable partial denture framework fabrication. The investment materials were subjected to microwave and conventional oven drying at different time intervals. MATERIAL AND METHODS: Type III partial denture casting investments (Wirovest, Wiroplus, Biosint, PH2) were tested. A total of 160 specimens were prepared in cylindrical form at a height of 40 mm and a diameter of 20 mm, in accordance with the manufacturers' recommendations. Forty specimens were prepared from each product; 20 specimens were dried at 230 degrees C for 1 hour in an electric furnace, the other 20 were dried in a microwave oven for 10 minutes at 600 W. The dried specimens were tested at 2- and 4-hour intervals in diametral compression at a crosshead speed of 0. 5 cm/min. RESULTS: The microwave drying technique resulted in greater diametral tensile strength values for all investment materials. CONCLUSION: Within the limitations of this study, microwave drying of type III dental investment materials at 600 W for 10 minutes was, apart from strengthening the material, timesaving for the dental laboratory. PMID- 10479255 TI - Comparison of accuracy between compression- and injection-molded complete dentures. AB - STATEMENT OF PROBLEM: A clinically significant incisal pin opening may occur after processing complete dentures if a compression molding technique is used. To recover the proper vertical dimension of occlusion, a time-consuming occlusal adjustment is necessary that often destroys the anatomy of the artificial teeth. A new injection molding process claims to produce dentures that require few, if any, occlusal adjustments in the laboratory after processing. PURPOSE: This laboratory study compared incisal pin opening, dimensional accuracy, and laboratory working time for dentures fabricated by this new injection system with dentures constructed by the conventional compression molding technique. MATERIAL AND METHODS: Two groups of 6 maxillary and 6 mandibular dentures were evaluated as follows: group 1 (control), Lucitone 199, compression molded with a long cure cycle; and group 2, Lucitone 199, injection molded with a long cure. Incisal pin opening was measured with a micrometer immediately after deflasking. A computerized coordinate measuring machine was used to measure dimensional accuracy of 3-dimensional variations in selected positions of artificial teeth in 4 stages of denture fabrication. Analysis of variance (ANOVA) and t tests were performed to compare the groups. RESULTS: A significant difference was found in pin opening between groups (t test). Horizontal dimensional changes evaluated with repeated measures ANOVA revealed no significant differences between groups. However, analysis of vertical dimensional changes disclosed significant differences between the groups. There was no appreciable difference in laboratory working time for flasking and molding denture bases between the injection and compression molding techniques when polymethyl methacrylate resin was used. CONCLUSION: The injection molding method produced a significantly smaller incisal pin opening over the standard compression molding technique. The injection molding technique, using polymethyl methacrylate, was a more accurate method for processing dentures. There were no appreciable differences in laboratory working time between the injection and compression molding techniques. PMID- 10479256 TI - Shock absorption potential of different mouth guard materials. AB - STATEMENT OF PROBLEM: Clinical stresses and situations in the mouths of patients are substantially different; thus, it is not possible to make an exact in vivo comparison of the efficiency of various mouth guards. PURPOSE: This study developed a device and a method to test and quantify the potential of mouth guards to absorb shock and evaluated and compared 5 designs for mouth guards. MATERIAL AND METHODS: Strain gauges and accelerometer sensors were mounted in various positions on the maxilla and inside an artificial skull. Sensors quantified the response of several areas of the skull to inputs of force on the maxillary teeth protected by the mouth guard being investigated. Input of force was applied to the maxillary teeth with a modal hammer equipped with a load cell. Five mouth guards of each of 5 designs were manufactured and placed in position before force was applied to the maxillary teeth. All inputs were measured and analyzed and, from those results, a method was developed to quantify the relative potential of the mouth guards to absorb shock. This method was then used to quantify the shock absorption potential of the 5 types of mouth guard designs. RESULTS: All 5 types of mouth guards provided some measure of protection and were better than no protection. However, the level of protection provided by the 5 mouth guard designs differed. PMID- 10479257 TI - Effects of smoking on implant success in grafted maxillary sinuses. AB - STATEMENT OF THE PROBLEM: Cigarette smoking has been identified as a significant risk factor for dental implant treatment. PURPOSE: This retrospective study evaluated the effect of smoking and the amount of cigarette consumption on the success rates of the implants placed in grafted maxillary sinuses. MATERIAL AND METHODS: Sixty patients (16 smokers and 44 nonsmokers) were evaluated for the effects of smoking on osseointegrated implants placed in 84 grafted maxillary sinuses that contained a total of 228 endosseous root-form implants. Seventy implants were placed in 26 maxillary sinuses in smokers, whereas 158 implants were placed in 58 sinuses in nonsmokers. The number of implant failures and the amount of cigarette consumption were recorded. RESULTS: Of the 228 implants, 205 (89.9%) remained in function, after a mean follow-up period of 41.6 months (2 to 60 months). There was a significantly higher cumulative implant success rate in nonsmokers (82.7%) than in smokers (65.3%) (P =.027). Overall cumulative implant success rate was 76%. There was no correlation between implant failures and the amount of cigarette consumption (P >.99). CONCLUSION: Within the limitations of this study, cigarette smoking appeared to be detrimental to the success of osseointegrated implants in grafted maxillary sinuses regardless of the amount of cigarette consumption. PMID- 10479258 TI - Effect of washers on reverse torque displacement of dental implant gold retaining screws. AB - STATEMENT OF THE PROBLEM: Gold screw loosening is a frequent problem that affects dental implants. PURPOSE: This study determined the effect of spring washers on gold screw displacements during applied removal torque in dental implants. MATERIAL AND METHODS: A 3-unit fixed partial denture was cast in semiprecious alloy and fixed to 2 dental implants with gold-retaining screws. Implants were embedded in acrylic resin and restrained from movement. A miniature load sensor was attached to a torque controller handpiece to accurately measure the time and torque needed to completely loosen the gold screws. Rotational displacements were calculated for 4 experimental setups, involving washers placed on (a) both, (b) the mesial, (c) the distal, and (d) no implants. Screw displacement data were compared between implants in all 4 combinations by means of paired t tests. RESULTS: Gold screws with underlying conical spring washers underwent on average up to 35% (14.1 microm) more rotational displacement during applied removal torque than those without washers. CONCLUSION: Inclusion of a conical spring washer significantly increased the amount of rotational displacement needed to completely loosen an implant gold retaining screw. PMID- 10479259 TI - Psychologic response of the edentulous patient after primary surgery for oral cancer: A cross-sectional study. AB - STATEMENT OF PROBLEM: Edentulous patients can have difficulty in tolerating dentures and this may lead to psychologic disturbance. The problem is potentially more severe for edentulous patients after primary surgery for oral cancer, where treatment can include composite resection and reconstruction, followed by adjuvant radiotherapy. PURPOSE: This study investigated the psychologic response and oral satisfaction of edentulous patients treated by surgery for oral squamous cell carcinoma, and to make a comparison to edentulous noncancer counterparts. METHODS AND MATERIAL: The cross-sectional study included patients who were alive and disease-free 2 to 3 years after primary surgery. Seventy patients underwent surgery at the Regional Maxillofacial Unit, Liverpool, in 1993 and 1994. Twenty eight patients were disease-free; 26 completed questionnaires that included a general health questionnaire (GHQ), a body satisfaction scale, a self-esteem scale, an oral symptom checklist, and a denture satisfaction questionnaire. Comparison was made with 98 noncancer edentulous patients from the same unit. RESULTS: There were similarities in psychologic and oral satisfaction scores between the noncancer and cancer edentulous patients. Cancer patients reported lower self-esteem (P <.02). Cancer patients who were not rehabilitated with either conventional or implant-retained prostheses had significant psychologic morbidity as measured by the GHQ, self-esteem, and body satisfaction scales. Cancer patients with implant-retained overdentures reported greater satisfaction with their dentures compared with their counterparts who wore conventional dentures (P <.05). CONCLUSION: Edentulous cancer patients who do not achieve oral rehabilitation after surgery for oral cancer exhibited significant psychologic morbidity. Patients with implant-retained overdentures exhibited a tendency to adopt the same psychologic response with improved denture satisfaction as edentulous patients with conventional dentures, despite the former having more extensive disease that would otherwise make the provision of dentures much more difficult if implants were not used. PMID- 10479260 TI - Trajectories of condylar points during nonworking side and protrusive movements of the mandible. AB - STATEMENT OF PROBLEM: During lateral excursive and protrusive jaw movements, condylar points are distant from any instantaneous rotational center. Therefore, it is likely that different condylar points would follow similar trajectories during these movements. PURPOSE: This study evaluated the effect of changes in condylar point location on trajectories of condylar points on the nonworking side and during a protrusive jaw movement and compared these changes with the effects described for open-close and working-side condylar movements in the same group of subjects. METHODS: The movements of 5 clinically determined condylar points were recorded in 44 subjects during a contralateral excursion and during protrusion (7 radiographically determined condylar points in 2 subjects). RESULTS: During any single jaw movement, the trajectory of each condylar point was similar in form and dimension to the other condylar points within that subject. CONCLUSION: Changes in condylar point location had little effect on the trajectories of condylar points on the nonworking side and during protrusive jaw movement. PMID- 10479261 TI - Disinfection of dental diamond burs contaminated with hepatitis B virus. AB - STATEMENT OF PROBLEM: Hepatitis B infection (HBV) is a significant hazard in the dental environment because the virus may be transmitted through contaminated dental instruments. PURPOSE: This study determined whether cold disinfectants can inactivate HBV DNA and HBV surface antigens on diamond burs contaminated with HBV and whether ultrasonication can increase the antiviral properties of these agents. MATERIAL AND METHODS: Sterile dental diamond burs were contaminated with serum from a patient who tested positive for HBV surface antigen and hepatitis B viral DNA. The burs were air dried and placed in solutions containing either Cidex, Asepsys, TBS, Rotagerm, Virkon disinfectants, or a control phosphate buffered saline. Burs were divided into 2 groups and disinfected for 15 minutes. The first group was ultrasonicated; the second group was not ultrasonicated during disinfection. All the burs were transferred to phosphate buffered saline and ultrasonicated to remove any remaining viral particles. The ultrasonicate was tested for the presence of HBV surface antigen with a microparticle enzyme immunoassay and for hepatitis B viral DNA with a chemiluminescent molecular hybridization assay. RESULTS: TBS did not require ultrasonication to inactivate viral DNA and surface antigen. Rotagerm and Virkon inactivated surface antigen and viral DNA only with ultrasonication. Cidex and Asepsys inactivated viral DNA but not surface antigen with ultrasonication. CONCLUSION: The chlorine containing compound TBS was the most active disinfectant tested and did not require ultrasonication to destroy HBV. The remaining disinfectants should be used with ultrasonication to inactivate HBV. PMID- 10479262 TI - Effectiveness of polymerization of a prosthetic composite using three polymerization systems. AB - STATEMENT OF PROBLEM: Although properties of laboratory-polymerized composite materials are influenced by the type of polymerizing unit, little information is available regarding the comparison between use of a high-intensity light source and application of secondary heat treatment. PURPOSE: This study examined properties of a prosthetic veneering composite polymerized with 3 polymerizing systems to evaluate the effects of varying polymerization modes on hardness, solubility, and depth of cure. MATERIAL AND METHODS: A composite material designed for a prosthetic veneer (Conquest Crown and Bridge) was polymerized using 3 methods: (1) exposure in the proprietary photopolymerizing unit with 2 halogen lamps (Cure-Lite Plus), followed by heating in an oven (Conquest Automatic Curing Unit); (2) exposure in a photopolymerizing unit with a xenon stroboscopic light source (Dentacolor XS); and (3) exposure in a photopolymerizing unit with 2 metal halide lamps (Hyper LII). Knoop hardness, water solubility, and depth of cure were determined for groups of 5 specimens, according to standardized testing methods. Data were compared using analysis of variance and the Duncan new multiple range test (P <.05). RESULT: The hardness number generated with the metal halide unit was statistically greater than those produced by the other 2 methods, and material component released into water was minimal when the material was exposed with the metal halide unit (P <.05). Among the 3 photopolymerizing units, the metal halide unit consistently exhibited the greatest depth of cure. CONCLUSION: Certain properties generated with the use of the high-intensity polymerizing unit exceeded those obtained from a proprietary system that requires a postheat treatment. PMID- 10479263 TI - Fatigue testing and microscopic evaluation of post and core restorations under artificial crowns. AB - STATEMENT OF PROBLEM: Coronoradicular reconstruction techniques of pulpless teeth include prefabricated post systems that retain a core material such as silver amalgam, composite, glass ionomer, or modified glass ionomer cement. Mechanical properties of these materials are critical to sustain masticatory forces. PURPOSE: This in vitro study compared the mechanical resistance of 3 core materials (silver amalgam, composite, and silver-reinforced glass ionomer) under masticatory conditions. MATERIAL AND METHODS: Industrially fabricated teeth were used and a total of 75 specimens were divided into 3 groups of 25 specimens. Titanium canal posts were placed, followed by core buildups in amalgam, composite, or glass ionomer. Teeth were prepared for full cast crowns and the crowns were fabricated and cemented with glass ionomer cement. Twenty specimens from each group were placed in a mastication simulator cyclically loading the teeth with a 400 N force for 1.5 million cycles. The 5 remaining specimens were used as controls. Teeth were sectioned and observed macroscopically and microscopically to determine the rate of defects for each material. Observed defects were verified with the Kruskal-Wallis test. The 3 core materials were ranked with the Tukey multiple comparison test. RESULTS: Significant differences of mechanical behavior were found for the 3 materials. At P <.01, silver amalgam was significantly superior to composite and glass ionomer. Composite was significantly superior to glass ionomer. CONCLUSION: Cores fabricated with amalgam had the lowest rate of defects when tested under artificial crowns. Glass ionomer, when used as a core material under artificial crowns, showed the highest rate of defects after an instantaneous load of 400 N for a 1,500,000 repetition cycle. PMID- 10479264 TI - Measurement of adsorption of salivary proteins onto soft denture lining materials. AB - STATEMENT OF PROBLEM: The adhesion of Candida albicans to soft liners is a major causative factor in denture stomatitis. It has been suggested that salivary proteins play an important role in this candidal adhesion. PURPOSE: This study measured the adsorption of salivary proteins on soft liners. MATERIAL AND METHODS: Five commercial materials and 1 experimental material were immersed in saliva, albumin solution, or milk. Proteins adsorbed on the material surfaces were measured by ATR/FT-IR spectroscopy. RESULTS: The amount of proteins adsorbed to the 6 materials varied considerably. Protein adsorption was significantly lower in the experimental fluoropolymer and polyphosphazene, and higher in acrylic resin and silicone. CONCLUSION: Different soft liners promote adsorption of varying amounts of protein. PMID- 10479265 TI - Attitude and disposition: Do they make a difference in cancer survival? AB - STATEMENT OF PROBLEM: Investigators have attempted to resolve the question: do psychosocial, emotional, and attitudinal factors have any effect on cancer survival? PURPOSE: This article presents a literature review and an overview of the developments in the mind-body connection to examine the influence of attitude in cancer survival. CONCLUSION: Even though a causal effect of emotional issues on the initiation and promotion of cancer is speculative, the effect of emotions on numerous medical conditions cannot be ignored. PMID- 10479266 TI - Programmed occlusal reconstruction in anticipation of tooth wear. AB - A procedure is presented that developed posterior lateral contacts in group function, despite existence of an anterior canine disclusion. This procedure allows a smooth transition into group function as the canine is subjected to natural attrition. PMID- 10479267 TI - Achieving an even thickness in heat-polymerized permanent acrylic resin denture bases for complete dentures. AB - Permanent denture bases form the fitting surface of a denture and are constructed on a master cast, in heat-polymerized acrylic resin. These bases are strong and rigid and demonstrate both the fit and retention of the final prosthesis. General recommendations suggests a thickness of between 1.5 to 3 mm for these bases. The normal procedure for producing such a denture base is to manually adapt materials to the cast, in their plastic state, before processing. Such procedures are liable to distortion and variation in the thickness of the resultant denture base and may be unreliable. This article describes a method for achieving a consistent thickness in a heat-polymerized, permanent, acrylic resin denture base through the use of a vacuum-formed, thermoplastic blank as a template. The method is simple, and results in a denture base with a consistent even thickness. PMID- 10479268 TI - Use of custom-made conformers in the treatment of ocular defects. AB - The loss of an eye is often followed by scar tissue contracture. The purpose of this article is to describe a procedure for fabricating acrylic resin ocular custom-made conformers to maintain socket size and contour. These custom-made ocular conformers act as an interim measure and as valuable diagnostic indicators of problems experienced by the patients. While stock conformers merely maintain the socket size and prevent scar tissue contractures, the custom-made conformers can also be used to enlarge unfavorably small sockets, stimulate eyelid movement, aid hygiene, help the clinician develop the final shape for the definitive prosthesis, and reduce the amount of postinsertion adjustments needed. These advantages and the ease of fabrication make custom-made conformers a viable initial treatment option in patients with ocular defects. PMID- 10479269 TI - Stabilizing record bases for edentulous obturator prostheses with silicone resilient relining material. AB - Stable record bases are critical for recording accurate maxillomandibular relations and in the evaluation of the esthetics and phonetics of wax trial prostheses. However, the record base for a maxillary obturator is often compromised because of a surgical defect and/or because the weight of the record base creates instability. This article describes a simple procedure for retaining and stabilizing a record base for an edentulous obturator prosthesis using silicone resilient relining materials. PMID- 10479270 TI - Use of the tongue for recording centric relation for edentulous patients. PMID- 10479271 TI - Altered guide pin as an abutment holder for handling an angulated abutment. PMID- 10479272 TI - Simple method of unscrewing tightened abutment and transfer coping in Calcitek Spline implants. PMID- 10479273 TI - Reply. PMID- 10479274 TI - Reply. PMID- 10479276 TI - Conceptually new sulfone analogues of the hormone 1alpha, 25-dihydroxyvitamin D(3): synthesis and preliminary biological evaluation. AB - A conceptually new series of vitamin D(3)-like nonfluorinated and fluorinated 16 ene side chain tert-butyl sulfones 3-7 has been synthesized. Even though these novel C,D-ring side chain analogues of the hormone 1alpha,25-dihydroxyvitamin D(3) (1,1,25D(3)) lack a terminal OH group, thought previously to be essential for high biological activity, they are highly antiproliferative and, in several cases, transcriptionally active in vitro but desirably noncalcemic in vivo. The side chain sulfone group may be binding to the nVDR as a hydrogen-bond acceptor, in contrast to the hydrogen-bond donor function of the 25-OH group of natural 1,25D(3). PMID- 10479275 TI - A multisubstrate adduct inhibitor of AICAR transformylase. PMID- 10479277 TI - Probing the binding of indolactam-V to protein kinase C through site-directed mutagenesis and computational docking simulations. AB - Protein kinase C (PKC) comprises a family of ubiquitous enzymes transducing signals by the lipophilic second messenger sn-1, 2-diacylglycerol (DAG). Teleocidin and its structurally simpler congener indolactam-V (ILV) bind to PKC with high affinity. In this paper, we report our computational docking studies on ILV binding to PKC using an automatic docking computer program, MCDOCK. In addition, we used site-directed mutagenesis to assess the quantitative contribution of crucial residues around the binding site of PKC to the binding affinity of ILV to PKC. On the basis of the docking studies, ILV binds to PKC in its cis-twist conformation and forms a number of optimal hydrogen bond interactions. In addition, the hydrophobic groups in ILV form "specific" hydrophobic interactions with side chains of a number of conserved hydrophobic residues in PKC. The predicted binding mode for ILV is entirely consistent with known structure-activity relationships and with our mutational analysis. Our mutational analysis establishes the quantitative contributions of a number of conserved residues to the binding of PKC to ILV. Taken together, our computational docking simulations and analysis by site-directed mutagenesis provide a clear understanding of the interaction between ILV and PKC and the structural basis for design of novel, high-affinity, and isozyme-selective PKC ligands. PMID- 10479279 TI - N-substituted adenosines as novel neuroprotective A(1) agonists with diminished hypotensive effects. AB - The synthesis and pharmacological profile of a series of neuroprotective adenosine agonists are described. Novel A(1) agonists with potent central nervous system effects and diminished influence on the cardiovascular system are reported and compared to selected reference adenosine agonists. The novel compounds featured are derived structurally from two key lead structures: 2-chloro-N-(1 phenoxy-2-propyl)adenosine (NNC 21-0041, 9) and 2-chloro-N-(1 piperidinyl)adenosine (NNC 90-1515, 4). The agonists are characterized in terms of their in vitro profiles, both binding and functional, and in vivo activity in relevant animal models. Neuroprotective properties assessed after postischemic dosing in a Mongolian gerbil severe temporary forebrain ischemia paradigm, using hippocampal CA1 damage endpoints, and the efficacy of these agonists in an A(1) functional assay show similarities to some reference adenosine agonists. However, the new compounds we describe exhibit diminished cardiovascular effects in both anesthetized and awake rats when compared to reference A(1) agonists such as (R) phenylisopropyladenosine (R-PIA, 5), N-cyclopentyladenosine (CPA, 2), 4, N [(1S,trans)-2-hydroxycyclopentyl]adenosine (GR 79236, 26), N-cyclohexyl-2'-O methyladenosine (SDZ WAG 994, 27), and N-[(2-methylphenyl)methyl]adenosine (Metrifudil, 28). In mouse permanent middle cerebral artery occlusion focal ischemia, 2-chloro-N-[(R)-[(2-benzothiazolyl)thio]-2-propyl]adenosine (NNC 21 0136, 12) exhibited significant neuroprotection at the remarkably low total intraperitoneal dose of 0.1 mg/kg, a dose at which no cardiovascular effects are observed in conscious rats. The novel agonists described inhibit 6, 7-dimethoxy-4 ethyl-beta-carboline-3-carboxylate-induced seizures, and in mouse locomotor activity higher doses are required to reach ED(50) values than for reference A(1) agonists. We conclude that two of the novel adenosine derivatives revealed herein, 12 and 5'-deoxy-5'-chloro-N-[4-(phenylthio)-1-piperidinyl]adenosine (NNC 21-0147, 13), representatives of a new series of P(1) ligands, reinforce the fact that novel selective adenosine A(1) agonists have potential in the treatment of cerebral ischemia in humans. PMID- 10479278 TI - Synthesis of novel GABA uptake inhibitors. 3. Diaryloxime and diarylvinyl ether derivatives of nipecotic acid and guvacine as anticonvulsant agents. AB - (3R)-1-[4,4-bis(3-methyl-2-thienyl)-3-butenyl]-3-piperidinecarboxylic acid 1 (tiagabine, Gabitril) is a potent and selective gamma-aminobutyric acid (GABA) uptake inhibitor with proven anticonvulsant efficacy in humans. This drug, which has a unique mechanism of action among marketed anticonvulsant agents, has been launched for add-on treatment of partial seizures with or without secondary generalization in patients >12 years of age. Using this new agent as a benchmark, we have designed two series of novel GABA uptake inhibitors of remarkable potency, using a putative new model of ligand interaction at the GABA transporter type 1 (GAT-1) uptake site. This model involves the postulated interaction of an electronegative region in the GABA uptake inhibitor with a positively charged domain in the protein structure of the GAT-1 site. These two novel series of anticonvulsant agents contain diaryloxime or diarylvinyl ether functionalities linked to cyclic amino acid moieties and were derived utilizing the new model, via a series of design steps from the known 4,4-diarylbutenyl GABA uptake inhibitors. The new compounds are potent inhibitors of [(3)H]-GABA uptake in rat brain synaptosomes in vitro, and their antiepileptic potential was demonstrated in vivo by their ability to protect against seizures induced by the benzodiazepine receptor inverse agonist methyl 4-ethyl-6,7-dimethoxy-beta carboline-3-carboxylate (DMCM) in mice. From structure-activity studies of these new GABA uptake inhibitors, we have shown that insertion of an ether oxygen in conjugation with the double bond in tiagabine (K(i) = 67 nM) improves in vitro potency by 5-fold to 14 nM. PMID- 10479280 TI - Synthesis of 1-(2-aminophenyl)isoquinolines and the biological activity of their cis-dichloro platinum(II) complexes. AB - The broad biological effects of isoquinolines prompted us to use them as chelating, nonleaving ligands in cis-platinum(II) antitumor complexes. The synthesis of several 1-(2-aminophenyl)isoquinoline derivatives with different levels of hydrogenation and varying substitution of the phenyl ring is reported. These compounds constitute a new class of ligands for the synthesis of oligocyclic platinum(II) complexes. In vitro cytotoxicity tests indicate that the most basic amine ligands afford the most effective complexes. Two of the new complexes were more potent against L1210 murine leukemia cells than the well established antitumor compound cisplatinum. PMID- 10479281 TI - Substituted analogues of GV150526 as potent glycine binding site antagonists in animal models of cerebral ischemia. AB - A series of analogues of the indole-2-carboxylate GV150526, currently in clinical trials as a potential neuroprotective agent for the control of the cerebral damage after stroke onset, was designed based on previous studies dealing with the electronic features of the north-east region of the glycine binding site associated with the NMDA receptor. In particular, the substitution of the para position of the terminal phenyl ring of GV150526 with suitable hydrophilic groups resulted in the identification of a new class of glycine antagonists. These compounds exhibited nanomolar in vitro affinity to the glycine binding site, high receptor selectivity, and outstanding in vivo potency. In particular, 3-[(E)-2 [(4-ureidomethylphenyl)aminocarbonyl]ethenyl]-4, 6-dichloroindole-2-carboxylic acid was found to be highly effective in the middle cerebral artery occlusion (MCAo) model in the rat, an animal model of focal ischemia, when given both prior to and after the occlusion of the middle cerebral artery. Notably, a significant neuroprotective effect was seen in this model postischaemia, when the administration of this compound was delayed up to 6 h from the occlusion of the middle cerebral artery, further confirming the wide therapeutic window seen for GV150526A. PMID- 10479282 TI - 8,11-dihydroxy-6-[(aminoalkyl)amino]-7H-benzo[e]perimidin-7-ones with activity in multidrug-resistant cell lines: synthesis and antitumor evaluation. AB - The synthesis of dihydroxybenzoperimidine derivatives, which are chromophore modified dihydroxyanthracenediones with an additional pyrimidine ring incorporated into the chromophore, is reported. These derivatives are structurally related to the antitumor agent mitoxantrone. Their synthesis was carried out by the reaction of 6-amino-8,11-dihydroxy-7H-benzo[e]perimidin-7-one (5) or 6,8, 11-trihydroxy-7H-benzo[e]perimidin-7-one (10) with a number of respective (alkylamino)alkylamines. The dihydroxybenzoperimidine derivatives exhibited in vitro cytotoxic activity against murine leukemia L1210 and human leukemia HL60 cell lines comparable to that of mitoxantrone. These compounds also exhibited a range of in vitro activity against the human MDR-type resistant leukemia K562R cell line with the MDR phenotype. The most active compound of this series, namely 6a, exhibited potent in vitro cytotoxic activity against a panel of human cell lines. Furthermore, in contrast to both mitoxantrone and doxorubicin, it displayed little cross-resistance in cell lines characterized by a MDR phenotype. Cell cycle analysis in the sensitive HT-29 and mitoxantrone resistant HT-29/Mx (not identified resistance mechanism) cell lines has revealed that both mitoxantrone and 6a induce a G2/M block. However, while the proportion of apoptotic cells after mitoxantrone treatment is similar for both sensitive and resistant cell lines, it is much lower for 6a. Compound 6a tested against P388 murine leukemia in vivo displayed a significant antitumor effect (%T/C 196 at an optimal dose of 10 mg/kg). The property of overcoming the cross-resistance was maintained also in in vivo efficacy studies, where no difference was observed in the antitumor activity of compound 6a against the A2780 human tumor xenograft and its MDR A2780/Dx subline. We conclude that benzoperimidines, if properly substituted, constitute a novel class of compounds that can overcome multidrug resistance. PMID- 10479283 TI - CoMFA study of novel phenyl ring-substituted 3alpha-(diphenylmethoxy)tropane analogues at the dopamine transporter. AB - A series of phenyl ring-substituted analogues of 3alpha-(diphenylmethoxy)tropane (benztropine) has been prepared as novel probes for the dopamine transporter. Cross-validated comparative molecular field analysis (CoMFA) models of the binding domain on the dopamine transporter were constructed using 37 geometry optimized structures of these compounds and their corresponding binding affinities (K(i) values) for the displacement of [(3)H]WIN 35,428 or potency of [(3)H]dopamine uptake inhibition (IC(50) values) in rat caudate putamen tissue. The most predictive model (q(2) = 0.78) correlated the steric component of CoMFA to the dependent variable of [(3)H]WIN 35,428 binding affinities. A novel series of seven phenyl ring-substituted analogues of 3alpha-(diphenylmethoxy)tropane was prepared, and our best molecular model was used to accurately predict their binding affinities. This study is the first to provide a CoMFA model for this class of dopamine uptake inhibitors. This model represents an advancement in the design of novel dopamine transporter ligands, based on 3alpha (diphenylmethoxy)tropane, and further substantiates structure-activity relationships that have previously been proposed for this class of compounds. This CoMFA model can now be used to predict the binding affinities of novel 3alpha-(diphenylmethoxy)tropane analogues at the dopamine transporter and will be useful in the design of molecular probes within this class of dopamine uptake inhibitors. PMID- 10479284 TI - 5-deazafolate analogues with a rotationally restricted glutamate or ornithine side chain: synthesis and binding interaction with folylpolyglutamate synthetase. AB - Rotationally restricted analogues of 5-deazapteroyl-L-glutamate and (6R,6S)-5 deaza-5,6,7,8-tetrahydropteroyl-L-glutamate with a one-carbon bridge between the amide nitrogen and the 6'-position of the p-aminobenzoyl moiety were synthesized and tested as substrates for folylpolyglutamate synthetase (FPGS), a key enzyme in folate metabolism and an important determinant of the therapeutic potency and selectivity of classical antifolates. The corresponding bridged analogues of 5 deazapteroyl-L-ornithine and (6R,6S)-5-deaza-5,6,7, 8-tetrahydropteroyl-L ornithine were also synthesized as potential inhibitors. Condensation of diethyl L-glutamate with methyl 2-bromomethyl-4-nitrobenzoate followed by catalytic reduction of the nitro group, reductive coupling with 2-acetamido-6 formylpyrido[2, 3-d]pyrimidin-4(3H)-one in the presence of dimethylaminoborane, and acidolysis with HBr/AcOH yielded 2-L-[5-[N-(2-acetamido-4(3H)-oxopyrido[2, 3 d]pyrimidin-6-yl)methylamino]-2, 3-dihydro-1-oxo-2(1H)-isoindolyl]glutaric acid (1). When acidolysis was preceded by catalytic hydrogenation, the final product was the corresponding (6R,6S)-tetrahydro derivative 2. A similar sequence starting from methyl N(delta)-benzyloxycarbonyl-L-ornithine led to 2-L-[5-[N-(2 amino-4(3H)-oxopyrido[2, 3-d]pyrimidin-6-yl)methylamino]-2, 3-dihydro-1-oxo-2(1H) isoindolyl]-5-aminopentanoic acid (3) and the (6R,6S)-tetrahydro derivative 4. Compounds 3 and 4 were powerful inhibitors of recombinant human FPGS, whereas 1 and 2 were exceptionally efficient FPGS substrates, with the reduced compound 2 giving a K(m) (0.018 microM) lower than that of any other substrate identified to date. (6R,6S)-5-Deazatetrahydrofolate, in which the side chain is free to rotate, was rapidly converted to long-chain polyglutamates. In contrast, the reaction of 1 and 2 was limited to the addition of a single molecule of glutamic acid. Hence rotational restriction of the side chain did not interfere with the initial FPGS catalyzed reaction and indeed seemed to facilitate it, but the ensuing gamma glutamyl adduct was no longer an efficient substrate for the enzyme. PMID- 10479285 TI - The opioid mu agonist/delta antagonist DIPP-NH(2)[Psi] produces a potent analgesic effect, no physical dependence, and less tolerance than morphine in rats. AB - Opioid compounds with mixed mu agonist/delta antagonist properties are expected to be analgesics with low propensity to produce tolerance and dependence. In an effort to strengthen the mu agonist component of the mixed mu agonist/delta antagonist H-Tyr-Tic-Phe-Phe-NH(2) (TIPP-NH(2)), analogues containing structurally modified tyrosine residues in place of Tyr(1) were synthesized. Among the prepared compounds, H-Dmt-Tic-Phe-Phe-NH(2) (DIPP-NH(2); Dmt = 2',6' dimethyltyrosine) and H-Dmt-TicPsi[CH(2)NH]Phe-Phe-NH(2) (DIPP-NH(2)[Psi]) retained a mixed mu agonist/delta antagonist profile, as determined in the guinea pig ileum and mouse vas deferens assays, whereas H-Tmt-Tic-Phe-Phe-NH(2) (Tmt = N,2',6'-trimethyltyrosine) was a partial mu agonist/delta antagonist and H-Tmt TicPsi[CH(2)NH]Phe-Phe-NH(2) was a mu antagonist/delta antagonist. DIPP NH(2)[Psi] showed binding affinities in the subnanomolar range for both mu and delta receptors in the rat brain membrane binding assays, thus representing the first example of a balanced mu agonist/delta antagonist with high potency. In the rat tail flick test, DIPP-NH(2)[Psi] given icv produced a potent analgesic effect (ED(50) = 0.04 microg), being about 3 times more potent than morphine (ED(50) = 0.11 microg). It produced less acute tolerance than morphine but still a certain level of chronic tolerance. Unlike morphine, DIPP-NH(2)[Psi] produced no physical dependence whatsoever upon chronic administration at high doses (up to 4.5 microg/h) over a 7-day period. In conclusion, DIPP-NH(2)[Psi] fulfills to a large extent the expectations based on the mixed mu agonist/delta antagonist concept with regard to analgesic activity and the development of tolerance and dependence. PMID- 10479286 TI - Synthesis, opioid receptor binding, and biological activities of naltrexone derived pyrido- and pyrimidomorphinans. AB - A series of pyrido- and pyrimidomorphinans (6a-h and 7a-g) were synthesized from naltrexone and evaluated for binding and biological activity at the opioid receptors. The unsubstituted pyridine 6a displayed high affinities at opioid delta, mu, and kappa receptors with K(i) values of 0.78, 1.5, and 8.8 nM, respectively. Compound 6a was devoid of agonist activity in the mouse vas deferens (MVD) and guinea pig ileum (GPI) preparations but was found to display moderate to weak antagonist activity in the MVD and GPI with K(e) values of 37 and 164 nM, respectively. The pyrimidomorphinans in general displayed lower binding potencies and delta receptor binding selectivities than their pyridine counterparts. Incorporation of aryl groups as putative delta address mimics on the pyrido- and pyrimidomorphinan framework gave ligands with significant differences in binding affinity and intrinsic activity. Attachment of a phenyl group at the 4'-position of 6a or the equivalent 6'-position of 7a led to dramatic reduction in binding potencies at all the three opioid receptors, indicating the existence of a somewhat similar steric constraint at the ligand binding sites of delta, mu, and kappa receptors. In contrast, the introduction of a phenyl group at the 5'-position of 6a did not cause any reduction in the binding affinity at the delta receptor. In comparison to the unsubstituted pyridine 6a, the 5'-phenylpyridine 6c showed improvements in mu/delta and kappa/delta binding selectivity ratios as well as in the delta antagonist potency in the MVD. Interestingly, introduction of a chlorine atom at the para position of the pendant 5'-phenyl group of 6c not only provided further improvements in delta antagonist potency in the MVD but also shifted the intrinsic activity profile of 6c from an antagonist to that of a mu agonist in the GPI. Compound 6d thus possesses the characteristics of a nonpeptide mu agonist/delta antagonist ligand with high affinity at the delta receptor (K(i) = 2.2 nM), high antagonist potency in the MVD (K(e) = 0.66 nM), and moderate agonist potency in the GPI (IC(50) = 163 nM). Antinociceptive evaluations in mice showed that intracerebroventricular (icv) injections of 6d produced a partial agonist effect in the 55 degrees C tail-flick assay and a full agonist effect in the acetic acid writhing assay (A(50) = 7.5 nmol). No signs of overt toxicity were observed with this compound in the dose ranges tested. Moreover, repeated icv injections of an A(90) dose did not induce any significant development of antinociceptive tolerance in the acetic acid writhing assay. The potent delta antagonist component of this mixed mu agonist/delta antagonist may be responsible for the diminished propensity to produce tolerance that this compound displays. PMID- 10479287 TI - Synthesis, biological activity, and conformational analysis of four seco-D-15,19 bisnor-1alpha,25-dihydroxyvitamin D analogues, diastereomeric at C17 and C20. AB - The synthesis of four CD-ring-modified 19-nor-1alpha, 25-dihydroxyvitamin D(3) derivatives lacking C15, referred to as 6C analogues, and diastereomeric at C17 and C20 is described. The synthesis involves an Ireland-Claisen rearrangement of a 3-methyl-substituted ester of (1R)-3-methyl-2-cyclohexen-1-ol as the key step, followed by elaboration of the side chain, transformation into a C8 cyclohexanone derivative, and final Wittig-Horner coupling with the 19-nor A-ring phosphine oxide. Despite possessing a more flexible side chain than the parent hormone, biological evaluation showed an unexpected superagonistic antiproliferative and prodifferentiating activity (10-50 times higher as compared to that of 1alpha,25(OH)(2)D(3)) for the diastereomer with the "natural" configuration at C17 and C20. The other diastereomers exhibit a 25-90% decrease in activity. All four analogues show a decreased binding affinity (45% or less), and their calcemic activity is 4-400 times less than that of 1alpha,25(OH)(2)D(3). The conformational behavior of their side chain was studied using molecular mechanics calculations, and the result is presented as volume maps. A relative activity volume was determined by subtraction of the volume map of the least active analogue from the volume map of the most active one. This shows three regions corresponding to preferred orientations in space of the side chain of the active analogue. One of these regions was found to overlap with the region that is preferentially occupied by the most active of the four diastereomeric 22-methyl substituted 1alpha,25(OH)(2)D(3) analogues. PMID- 10479288 TI - Design and structure-activity relationships of potent and selective inhibitors of blood coagulation factor Xa. AB - The discovery of a series of non-peptide factor Xa (FXa) inhibitors incorporating 3-(S)-amino-2-pyrrolidinone as a central template is described. After identifying compound 4, improvements in in vitro potency involved modifications of the liphophilic group and optimizing the angle of presentation of the amidine group to the S1 pocket of FXa. These studies ultimately led to compound RPR120844, a potent inhibitor of FXa (K(i) = 7 nM) which shows selectivity for FXa over trypsin, thrombin, and several fibrinolytic serine proteinases. RPR120844 is an effective anticoagulant in both the rat model of FeCl(2)-induced carotid artery thrombosis and the rabbit model of jugular vein thrombus formation. PMID- 10479289 TI - Sulfonamidopyrrolidinone factor Xa inhibitors: potency and selectivity enhancements via P-1 and P-4 optimization. AB - Sulfonamidopyrrolidinones were previously disclosed as a selective class of factor Xa (fXa) inhibitors, culminating in the identification of RPR120844 as a potent member with efficacy in vivo. Recognizing the usefulness of the central pyrrolidinone template for the presentation of ligands to the S-1 and S-4 subsites of fXa, studies to optimize the P-1 and P-4 groups were initiated. Sulfonamidopyrrolidinones containing 4-hydroxy- and 4-aminobenzamidines were discovered to be effective inhibitors of fXa. X-ray crystallographic experiments in trypsin and molecular modeling studies suggest that our inhibitors bind by insertion of the 4-hydroxybenzamidine moiety into the S-1 subsite of the fXa active site. Of the P-4 groups examined, the pyridylthienyl sulfonamides were found to confer excellent potency and selectivity especially in combination with 4-hydroxybenzamidine. Compound 20b (RPR130737) was shown to be a potent fXa inhibitor (K(i) = 2 nM) with selectivity against structurally related serine proteinases (>1000 times). Preliminary biological evaluation demonstrates the effectiveness of this inhibitor in common assays of thrombosis in vitro (e.g. activated partial thromboplastin time) and in vivo (e.g. rat FeCl(2)-induced carotid artery thrombosis model). PMID- 10479290 TI - Synthesis and biochemical evaluation of 3-fluoromethyl-1,2,3, 4 tetrahydroisoquinolines as selective inhibitors of phenylethanolamine N methyltransferase versus the alpha(2)-adrenoceptor. AB - A series of 3-fluoromethyl-1,2,3,4-tetrahydroisoquinolines (3-fluoromethyl-THIQs) was proposed, and their phenylethanolamine N-methyltransferase (PNMT) and alpha(2)-adrenoceptor affinities were predicted through the use of comparative molecular field analysis (CoMFA) models. These compounds were synthesized and evaluated for affinity at PNMT and the alpha(2)-adrenoceptor. It was discovered that these compounds are some of the most selective inhibitors of PNMT versus the alpha(2)-adrenoceptor known. To determine the ability of these compounds to penetrate the blood-brain barrier (BBB), a series of THIQs possessing a variety of calculated partition coefficients (Clog P) were assayed using an in vitro BBB model. This study found a good correlation between lipophilicity (Clog P) and BBB permeability, which indicated that THIQs possessing Clog P values of at least 0.13-0.57 should have some penetration into the brain. Two compounds [3 fluoromethyl-7-N-(4-chlorophenyl)aminosulfonyl-THIQ (18) and 3-fluoromethyl-7 cyano-THIQ (20)] possess calculated partition coefficients greater than 0.57 and display selectivities (alpha(2)-adrenoceptor K(i)/PNMT K(i)) greater than 200 and thus represent promising leads in the development of highly selective inhibitors of PNMT with the ability to penetrate the BBB. PMID- 10479291 TI - Synthesis, structure-activity relationships, and RARgamma-ligand interactions of nitrogen heteroarotinoids. AB - Three heteroarotinoids containing a nitrogen atom in the first ring and a C-O linking group between the two aryl rings were synthesized and evaluated for RAR and RXR retinoid receptor transactivation, tumor cell growth inhibition, and transglutaminase (TGase) induction. Ethyl 4-(N,4,4-trimethyl-1,2,3,4 tetrahydroquinolinyl)benzoate (1) contained an N-CH(3) group and activated all retinoid receptors except for RARgamma. Inceasing the hydrophobicity around the rings with analogues ethyl 4-(N,4,4,7-tetramethyl-1,2,3, 4-tetrahydroquinolin-6 oyloxy)benzoate (2) [7-methyl group added] and ethyl 4-(4,4-dimethyl-N-isopropyl 1,2,3, 4-tetrahydroquinolin-6-oyloxy)benzoate (3) [NCH(CH(3))(2) group at C-4] increased the potency and specificity for RARalpha, RARbeta, and RXRalpha, compared to 1, but had little effect on RXRbeta and RXRgamma activation. Although 1 and 3 were unable to activate RARgamma, 2 did activate this receptor with efficacy and high potency equal to that of 9-cis-retinoic acid (9-c-RA). All three heteroarotinoids exhibited 5-8-fold greater specificities for RARbeta over RARalpha. In addition, esters 1-3 inhibited the growth of two cell lines each derived from cervix, vulvar, ovarian, and head/neck tumors with similar efficiencies to that of 9-c-RA through a mechanism independent of apoptosis. The vulvar cell lines were the most sensitive, and the ovarian lines were the least sensitive. Ester 2 was similar to 1 and 3 except that 2 was a much more potent growth inhibitor of the two vulvar cell lines, which is consistent with strong RARgamma activation by 2 (but not by 1 and 3) and the high levels of RARgamma expression in skin. All three heteroarotinoids induced production of TGase, a marker of retinoid activity in human erythroleukemic cells. Esters 2 and 3 were the more potent TGase activators than 1, in agreement with the stronger activation of the RAR receptors by 2 and 3. The biological activities of these agents, and the RARgamma potency of 2 in particular, demonstrate the promise of these compounds as pharmaceutics for cancer and skin disorders. PMID- 10479292 TI - Synthesis and cytotoxic evaluation of cycloheximide derivatives as potential inhibitors of FKBP12 with neuroregenerative properties. AB - On the basis of the new finding that the protein synthesis inhibitor cycloheximide (1, 4-[2-(3, 5-dimethyl-2-oxocyclohexyl)-2-hydroxyethyl]-2,6 piperidinedione) is able to competitively inhibit hFKBP12 (K(i) = 3.4 microM) and homologous enzymes, a series of derivatives has been synthesized. The effect of the compounds on the activity of hFKBP12 and their cytotoxicity against eukaryotic cell lines (mouse L-929 fibroblasts, K-562 leukemic cells) were determined. As a result, several less toxic or nontoxic cycloheximide derivatives were identified by N-substitution of the glutarimide moiety and exhibit IC(50) values in the range of 22.0-4.4 microM for inhibition of hFKBP12. Among these compounds cycloheximide-N-(ethyl ethanoate) (10, K(i) = 4.1 microM), which exerted FKBP12 inhibition to an extent comparable to that of cycloheximide (1), was found to cause an approximately 1000-fold weaker inhibitory effect on eukaryotic protein synthesis (IC(50) = 115 microM). Cycloheximide-N-(ethyl ethanoate) (10) was able to significantly speed nerve regeneration in a rat sciatic nerve neurotomy model at dosages of 30 mg/kg. PMID- 10479293 TI - Design and synthesis of water-soluble glucuronide derivatives of camptothecin for cancer prodrug monotherapy and antibody-directed enzyme prodrug therapy (ADEPT). AB - Glucuronide prodrugs of 9-aminocamptothecin were synthesized. Prodrug 4, in which 9-aminocamptothecin was connected to glucuronic acid by an aromatic spacer via a carbamate linkage, was stable in both aqueous solution and human plasma. Prodrug 4 and its potassium salt 12 were 20-80-fold less toxic than 9-aminocamptothecin to human tumor cell lines. The simultaneous addition of beta-glucuronidase and 4 or 12 to tumor cells resulted in a cytotoxic effect equal to that of 9 aminocamptothecin alone. Prodrugs 4 and 12 were over 80 and 4000 times more soluble than 9-aminocamptothecin in aqueous solutions at pH 4.0, respectively. Compounds 4 and 12 may be useful for prodrug monotherapy of tumors that accumulate extracellular lysosomal beta-glucuronidase as well as for antibody directed enzyme prodrug therapy (ADEPT) of cancer. PMID- 10479294 TI - Allosteric modulation of the adenosine A(1) receptor. Synthesis and biological evaluation of novel 2-amino-3-benzoylthiophenes as allosteric enhancers of agonist binding. AB - Novel allosteric enhancers of agonist binding to the rat adenosine A(1) receptor are described. The lead compound for the new series was PD 81,723 ((2-amino-4, 5 dimethyl-3-thienyl)[3-(trifluoromethyl)phenyl]methanone), a compound previously reported by Bruns and co-workers (Mol. Pharmacol. 1990, 38, 950-958). The 4,5 dimethyl group and the benzoyl moiety were targets for further modifications, leading to series of 4, 5-dialkyl (12a-g), of tetrahydrobenzo (12h-u), and of tetrahydropyridine (13a-g) derivatives. A number of compounds, in particular 12b, 12e, 12j, 12n, and 12u, proved superior to PD 81,723. Their EC(50) values for enhancing the binding of the adenosine A(1) receptor agonist N(6) cyclopentyladenosine to the receptor were lower, and/or their antagonistic activity on the adenosine A(1) receptor was shown to be diminished. PMID- 10479295 TI - 2-thioether 5'-O-(1-thiotriphosphate)adenosine derivatives as new insulin secretagogues acting through P2Y-Receptors. AB - P2-Receptors (P2-Rs) represent significant targets for novel drug development. P2 Rs were identified also on pancreatic B cells and are involved in insulin secretion. Therefore, novel P2Y-R ligands, 2-thioether 5'-O-phosphorothioate adenosine derivatives (2-RS-ATP-alpha-S), were synthesized as potential insulin secretagogues. An efficient synthesis of these nucleotides and a facile method for separation of the chiral products are described. The enzymatic stability of the compounds toward pig pancreas type I ATPDase was evaluated. The rate of hydrolysis of 2-hexylthio-5'-O-(1-thiotriphosphate)adenosine (2-hexylthio-ATP alpha-S) isomers by ATPDase was 28% of that of ATP. Some 2-thioether 5' (monophosphorothioate)adenosine derivatives (2-RS-AMP-S) exerted an inhibitory effect on ATPDase. The apparent affinity of the compounds to P2Y(1)-R was determined by measurement of P2Y-R-promoted phospholipase C activity in turkey erythrocyte membranes. 2-RS-ATP-alpha-S derivatives were agonists, stimulating the production of inositol phosphates with K(0.5) values in the nanomolar range. 2-RS-AMP-S derivatives were full agonists, although 2 orders of magnitude less potent. All the compounds were more potent than ATP. The effect on insulin secretion and pancreatic flow rate was evaluated on isolated and perfused rat pancreas. A high increase, up to 500%, in glucose-induced insulin secretion was due to addition of 2-hexylthio-ATP-alpha-S in the nanomolar concentration range, which represents 100-fold enhancement of activity relative to ATP. 2-Hexylthio AMP-S was 2.5 orders of magnitude less effective. PMID- 10479296 TI - Design, synthesis, and biological evaluation of novel non-piperazine analogues of 1-[2-(diphenylmethoxy)ethyl]- and 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3 phenylpropyl)piperazines as dopamine transporter inhibitors. AB - A series of novel diamine, amine-amide, and piperazinone analogues of N-[2 (bisarylmethoxy)ethyl]-N'-(phenylpropyl)piperazines, GBR 12909 and 12935, were synthesized and evaluated as inhibitors of presynaptic monoamine neurotransmitter transporters. The primary objective of the study was to determine the structural requirements for selectivity of ligand binding and potency for neurotransmitter reuptake inhibition. In general, the target compounds retained transporter affinity; however, structural variations produced significant effects on reuptake inhibition and transporter selectivity. For example, analogues prepared by replacing the piperazine ring in the GBR structure with an N, N' dimethylpropyldiamine moiety displayed enhanced selectivity for binding and reuptake inhibition at the norepinephrine (NE) transporter site (e.g. 4 and 5). Congeners in which the amide nitrogen atom was attached to the aralkyl moiety of the GBR molecule showed moderate affinity (K(i) = 51-61 nM) and selectivity for the dopamine transporter (DAT) site. In contrast, introduction of a carbonyl group adjacent to either nitrogen atom of the piperazine ring (e.g. 25 and 27) was not well tolerated. From the compounds prepared, analogue 16 was selected for further evaluation. With this congener, locomotor activity induced by cocaine at a dose of 20 mg/kg was attenuated with an AD(50) (dose attenuating cocaine induced stimulation by 50%) of 60.0 +/- 3.6 mg/kg. PMID- 10479297 TI - Drug delivery systems employing 1,4- or 1,6-elimination: poly(ethylene glycol) prodrugs of amine-containing compounds. AB - A general methodology for synthesizing poly(ethylene glycol) (PEG) prodrugs of amino-containing compounds has been developed and constitutes the basis for solubilization of insoluble drugs, extending plasma circulating half-lives and, in the case of anticancer agents, apparent tumor accumulation. Thus, we have successfully designed PEG conjugated specifiers or "triggers" as part of a double prodrug strategy that relies, first, on enzymatic separation of PEG followed by the classical and rapid 1,4- or 1, 6-benzyl elimination reaction releasing the amine (drug) bound in the form of a carbamate. The prodrug trigger was comprised of ester, carbonate, carbamate, or amide bonds in order to secure predictable rates of hydrolysis. Further refinement of the hydrolysis was accomplished by the introduction of steric hindrance through the use of ortho substituents on the benzyl component of the prodrug. This modification led to longer circulating plasma half-lives of the final tripartate form. The "ortho" effect also had the beneficial effect of directing nucleophilic attack almost exclusively to the activated benzyl 6-position of the heterobifunctional intermediates. In vivo testing of the PEG daunorubicin prodrugs (transport forms) prepared in the course of this study ultimately identified the type 1 carbamate (34b), with a circulating t(1/2) of 4 h, as the most effective derivative for solid tumor growth inhibition. PMID- 10479298 TI - Pyrrolidine-3-carboxylic acids as endothelin antagonists. 4. Side chain conformational restriction leads to ET(B) selectivity. AB - When the dialkylacetamide side chain of the ET(A)-selective antagonist ABT-627 is replaced with a 2,6-dialkylacetanilide, the resultant analogues show a complete reversal of receptor selectivity, preferring ET(B) over ET(A). By optimizing the aniline substitution pattern, as well as the alkoxy group on the 2-aryl substituent, it is possible to prepare antagonists with subnanomolar affinity for ET(B) and with selectivities in excess of 4000-fold. A number of these compounds also show promising pharmacokinetic profiles; a useful balance of properties is found in A-192621 (38). Pharmacology studies with A-192621 serve to reveal the role of the ET(B) receptor in modulating blood pressure; the observed hypertensive response to persistent ET(B) blockade is consistent with previous postulates and indicates that ET(B)-selective antagonists may not be suitable as agents for long-term systemic therapy. PMID- 10479299 TI - Design, synthesis, and activity of a series of pyrrolidine-3-carboxylic acid based, highly specific, orally active ET(B) antagonists containing a diphenylmethylamine acetamide side chain. AB - The endothelin (ET)-B receptor subtype is expressed on vascular endothelial and smooth muscle cells and mediates both vasodilation and vasoconstriction. On the basis of the pharmacophore of the previously reported ET(A)-specific antagonist 1, (ABT-627), we are reporting the discovery of a novel series of highly specific, orally active ET(B) receptor antagonists. Replacing the dibutylaminoacetamide group of 1 with a diphenylmethylaminoacetamide group resulted in antagonist 2 with a complete reversal of receptor specificity. Structure-activity relationship studies revealed that ortho-alkylation of the phenyl rings could further increase ET(B) affinity and also boost the ET(A)/ET(B) activity ratio of the resulting antagonists. A similar antagonism selectivity profile could also be achieved when one of the phenyl rings of the acetamide side chain was replaced with an alkyl group, preferably a tert-butyl group (10h). Combining these features with modification of the 2-aryl group of the pyrrolidine core, we have identified a potent antagonist (9k, A-308165) with over 27 000-fold selectivity favoring the ET(B) receptor and an acceptable pharmacokinetic profile (F = 24%) in rats. PMID- 10479300 TI - Carbonic anhydrase inhibitors: synthesis of water-soluble, aminoacyl/dipeptidyl sulfonamides possessing long-lasting intraocular pressure-lowering properties via the topical route. AB - Reaction of 26 aromatic/heterocyclic sulfonamides containing amino, imino, hydrazino, or hydroxyl groups with Boc-Gly, Boc-Sar, TrS-Crt, or Boc-Gly-Gly (Sar = sarcosine, N-Me-Gly; Crt = creatine, N-amidinosarcosine; TrS = tritylsulfenyl; Boc = tert-butoxycarbonyl) in the presence of carbodiimide derivatives afforded after removal of the protecting groups a series of water-soluble compounds (as salts of strong acids, such as hydrochloric, trifluoroacetic, or trifluoromethanesulfonic). The new derivatives were assayed as inhibitors of the zinc enzyme carbonic anhydrase (CA) and more precisely of three of its isozymes, CA I, II (cytosolic forms), and IV (membrane-bound form), involved in important physiological processes. Efficient inhibition was observed against all three isozymes and especially against CA II and IV (in the nanomolar range), the two isozymes known to play a critical role in aqueous humor secretion within the ciliary processes of the eye. Some of the best inhibitors synthesized were applied as 2% water solutions into the eye of normotensive or glaucomatous albino rabbits, when strong and long-lasting intraocular pressure (IOP) lowering was observed with many of them. Thus, the aminoacyl/dipeptidyl tail conferring water solubility to these sulfonamide CA inhibitors coupled with strong enzyme inhibitory properties and balanced lipid solubility seem to be the key factors for obtaining compounds with effective topical antiglaucoma activity. PMID- 10479301 TI - Second-generation peptidomimetic inhibitors of protein farnesyltransferase demonstrating improved cellular potency and significant in vivo efficacy. AB - The synthesis and evaluation of analogues of previously reported farnesyltransferase inhibitors, pyridyl benzyl ether 3 and pyridylbenzylamine 4, are described. Substitution of 3 at the 5-position of the core aryl ring resulted in inhibitors of equal or less potency against the enzyme and decreased efficacy in a cellular assay against Ras processing by the enzyme. Substitution of 4 at the benzyl nitrogen yielded 26, which showed improved efficacy and potency and yet presented a poor pharmacokinetic profile. Further modification afforded 30, which demonstrated a dramatically improved pharmacokinetic profile. Compounds 26 and 29 demonstrated significant in vivo efficacy in nude mice inoculated with MiaPaCa-2, a human pancreatic tumor-derived cell line. PMID- 10479302 TI - Cardioselective antiischemic ATP-sensitive potassium channel (K(ATP)) openers. 6. Effect of modifications at C6 of benzopyranyl cyanoguanidines. AB - The effect on potency and selectivity of modifications at the C6 position of the cardioprotective K(ATP) opener BMS-180448 (2) is described. Structure-activity studies show that a variety of electron-withdrawing groups (ketone, sulfone, sulfonamide, etc.) are tolerated for cardioprotective activity as measured by EC(25) values for an increase in time to the onset of contracture in globally ischemic rat hearts. Changes made to the sulfonamido substituent indicate that compounds derived from secondary lipophilic amines are preferred for good cardioprotective potency and selectivity. The diisobutyl analogue 27 (EC(25) = 0.04 microM) is the most potent compound of this series. The cardiac selectivity of 27 results from a combination of reduced vasorelaxant potency and enhanced cardioprotective potency relative to the potent vasodilating K(ATP) openers (e.g., cromakalim). The diisobutylsulfonamide analogue 27 is over 4 orders of magnitude more cardiac selective than cromakalim (1). These results support the hypothesis that the cardioprotective and vasorelaxant properties of K(ATP) openers follow distinct structure-activity relationships. The mechanism of action of 27 appears to involve opening of the cardiac K(ATP) as its cardioprotective effects are abolished by the K(ATP) blocker glyburide. PMID- 10479303 TI - Design, synthesis, and evaluation of chromen-2-ones as potent and selective human dopamine D4 antagonists. AB - The discovery of a series of chromen-2-ones with selective affinity for the dopamine (DA) D4 receptor is described. Target compounds were tested for binding to cloned human DA D2L, D3, and D4.2 receptor subtypes expressed in Chinese hamster ovary K1 cells. Several compounds demonstrated high affinity (<20 nM, K(i)) and greater than 100-fold selectivity for DA D4.2 versus DA D2L receptors. The results of a SAR study are discussed within. In a DA D4 functional assay measuring [(3)H]thymidine uptake, target compounds showed antagonist activity at the D4.2 receptor. Compound 22, 7-[(2-phenylaminoethylamino)methyl]chromen-2-one, increased DOPA (L-3,4-dihydroxyphenylalanine) accumulation 51% in the hippocampus and 23% in the striatum of rat brains when dosed orally at 20 mg/kg. PMID- 10479304 TI - Lobeline: structure-affinity investigation of nicotinic acetylcholinergic receptor binding. AB - (-)Lobeline (1) and (-)nicotine (2) bind at neuronal nicotinic cholinergic (nACh) receptors with high affinity (K(i) = 4 and 2 nM, respectively). Previous attempts to determine whether lobeline fits the currently accepted nicotinic pharmacophore model have led to suggestions that the carbonyl function, rather than the hydroxyl group, is a major contributor to binding. Interestingly, however, it has never been empirically demonstrated that either oxygen function is actually required for interaction with the receptor. In the present investigation we systematically examined a number of abbreviated analogues of lobeline and found that removal of either one or both oxygen functions reduces the affinity of lobeline by at least 25-fold; furthermore, oxidation of the (-)lobeline hydroxyl group (to afford lobelanine) or reduction of the carbonyl group (to afford lobelanidine) also resulted in decreased affinity. Although it is likely that both oxygen functions contribute to the high affinity of (-)lobeline at nACh receptors, it is concluded that the presence of both oxygen functions is not a requirement for binding; that is, replacement of the (-)lobeline hydroxyl group with a chloro group had no effect on affinity. Another finding of the present investigation is that removal of either one or both oxygen functions of lobeline results in compounds that retain the analgesic activity and potency of ( )lobeline, indicating that there is no direct relationship between neuronal nicotinic cholinergic (primarily alpha(4)beta(2) type) receptor affinity and spinal analgesia as measured in the tail-flick assay. PMID- 10479305 TI - IFN-gamma-derived lipopeptides: influence of lipid modification on the conformation and the ability to induce MHC class II expression on murine and human cells. AB - Two truncated analogues of a previously identified lipopeptide agonist toward the IFN-gamma receptor were synthesized in an attempt to determine the minimal compound able to induce expression of MHC class II molecules on murine and human cells and to study the role of the lipid tail. Circular dichroism studies were used to probe the induced conformationnal changes. Our results indicate at least a double role for the lipid modification that contributes to the stabilization of helical organization of the associated peptide and to its passive delivery into the cytoplasm. The persistence of biological activity in a truncated peptide of half of the residues present in the lead compound suggests that the lipid tail could also contribute to the stabilization of the peptide-receptor binding through additional hydrophobic interactions. This study allowed to readjust the minimal requirements for intracellular IFN-gamma receptor stimulation. More generally, we suggest that lipidated analogues of functional peptides could be utilized for intracellular target validation in the drug discovery process. PMID- 10479307 TI - Acylated iridoid and phenylethanoid glycosides from the aerial parts of Scrophularia nodosa. AB - From the aerial parts of Scrophularia nodosa L. (Scrophulariaceae), eighteen acylated iridoid glycosides and nine phenylethanoid glycosides were isolated. The structures of eight new iridoid and two new phenylethanoid glycosides were elucidated on the basis of chemical and spectroscopic evidences. PMID- 10479308 TI - Synthesis and absolute configuration of (S)-(-)- and (R)-(+)-2, 3-dihydro-2-(1 methylethenyl)-6-methoxybenzofuran AB - Resolution of racemic 2,3-dihydro-2-carboxy-6-methoxybenzofuran (2) by recrystallizations of diastereomeric salts prepared with (S)-(-)-alpha methylbenzylamine and (R)-(+)-alpha-methylbenzylamine gave the starting materials for the four-step total syntheses of (S)-(-)-2,3-dihydro-2-(1-methylethenyl)-6 methoxybenzofuran (1a) and (R)-(+)-2,3-dihydro-2-(1-methylethenyl)-6 methoxybenzofuran (1b). Their absolute configuration was established by chemical correlation. PMID- 10479306 TI - Small peptides containing phosphotyrosine and adjacent alphaMe-phosphotyrosine or its mimetics as highly potent inhibitors of Grb2 SH2 domain. AB - A series of small peptides with the sequence mAZ-pTyr-Xaa-Asn-NH(2), where Xaa denotes alpha-methylphosphotyrosine or its carboxylic mimetics, were synthesized as inhibitors of the Grb2 SH2 domain. Peptide 3 with (alpha-Me)pTyr as Xaa has the highest affinity for Grb2 (K(d) = 3 +/- 1 nM) and exhibits to date the best inhibitory activity (IC(50) = 11 +/- 1 nM) to displace PSpYVNVQN-Grb2 interaction in an ELISA test. The lower affinities of peptides with (alpha-Me)Tyr, (alpha Me)Phe(4-CO(2)H), or (alpha-Me)Phe(4-CH(2)CO(2)H) as Xaa demonstrate the importance of a double charged phosphate group at the pY+1 position. Molecular modeling showed additional hydrogen bond interactions provided by the (alpha Me)pTyr residue with the Grb2 SH2 domain. These results thus show that the effect of hydrophobic pY+1 residues, initially put forth to increased the binding affinities, can be further enhanced by a (-Me)pTyr residue which has both hydrophobic and hydrophilic properties. PMID- 10479310 TI - Sesquiterpene lactones from neurolaena oaxacana AB - Twelve sesquiterpene lactones, two new (1 and 2) and 10 known neurolenin-type germacranolides and furanoheliangolides (3-12) were isolated from Neurolaena oaxacana, and their structures were elucidated by NMR and GC-MS analysis. The chemotaxonomic importance of these findings is discussed. As N. lobata is used against dysenteries, neurolenin B (4) and a mixture of the neurolenins C (5) and D (6) were tested against Entamoeba histolytica and Giardia intestinalis. PMID- 10479309 TI - Antihyperglycemic sesquiterpenes from Psacalium decompositum. AB - Psacalium decompositum was investigated for antihyperglycemic compounds using diabetic ob/ob mice as a model for type 2 diabetes. In vivo bioassay-guided fractionation of an aqueous extract from the roots of P. decompositum led to the isolation of two new eremophilanolides, 3-hydroxycacalolide (1a) and epi-3 hydroxycacalolide (1b). A 1:1 mixture of 1a/1b exhibited antihyperglycemic activity when tested at 1.09 mmol/kg in ob/ob mice. The known furanoeremophilanes, cacalone (2a) and epicacalone (2b), were also isolated from the aqueous extract and were inactive. The known furanoeremophilane, cacalol (3), was isolated from a CH2Cl2 extract of P. decompositum roots and possessed antihyperglycemic activity. The relative stereochemistry in 1a and 1b was assigned 3R,5S and 3S,5S, respectively, based on ROESY data, 3J H-H values, and molecular mechanics calculations. Complete 13C and 1H NMR chemical shifts were assigned for 1a, 1b, 2a, 2b, and 3, and several revisions in 13C NMR assignments for 2a and 3 were made. Results from the conformational analysis of 1a, 1b, 2a, and 2b indicate that each compound exists in one major conformation in solution with H3-12 in a pseudoaxial position. PMID- 10479311 TI - Orizabins V-VIII, tetrasaccharide glycolipids from the Mexican scammony root (Ipomoea orizabensis). AB - An extensive investigation of the so-called jalapin resinoid obtained from roots of the Mexican scammony, Ipomoea orizabensis, using high field NMR spectroscopy led to the characterization of six glycosides, including the known scammonins I (1) and II (2) and four new tetrasaccharides of jalapinolic acid, orizabins V VIII (3-6). All the isolates (1-6) were found to be weakly cytotoxic toward human oral epidermoid carcinoma (KB). PMID- 10479312 TI - Phenylvaleric acid and flavonoid glycosides from Polygonum salicifolium. AB - (3R)-O-beta-D-Glucopyranosyloxy-5-phenylvaleric acid (1), (3R)-O-beta-D glucopyranosyloxy-5-phenylvaleric acid n-butyl ester (2), and a new dihydrochalcone diglycoside 4'-O-[beta-D-glucopyranosyl-(1-->6) glucopyranosyl]oxy-2'-hydroxy-3', 6'-dimethoxydihydrochalcone (3), together with six known flavonoid glycosides [kaempferol-3-O-beta-D-glucopyranoside (= astragalin) (4), kaempferol-3-O-beta-D-galactopyranoside (5), quercetin-3-O-beta D-glucopyranoside (= isoquercitrin) (6), quercetin-3-O-beta-D-galactopyranoside (= hyperoside) (7), quercetin-3-O-(2''-O-galloyl)-beta-D-glucopyranoside (8), and quercetin-3-O-beta-D-glucuronopyranoside (9)] were isolated from the aerial parts of Polygonum salicifolium. The structure elucidation of the isolated compounds was performed by spectroscopic (UV, IR, ESI-MS, 1D- and 2D-NMR), chemical (methylation, enzymatic hydrolysis, partial synthesis), and chromatographic methods (HPLC, Chiralcel OD). The flavonoid glycosides (4-9) demonstrated scavenging properties toward the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical in TLC autographic assays. PMID- 10479313 TI - Semisynthesis and cytotoxicity of styryl-lactone derivatives. AB - The cytotoxicity and the cell-cycle action of altholactone (1), goniofufurone (2), and eight altholactone derivatives (5-12), were determined in vitro on L 1210 cells. Semisyntheses and structure-activity relationships of these compounds are described. The results of this study suggest that the cytotoxicity of altholactone (1), 11-nitro-altholactone (8), and 7-chloro-6,7-dihydroaltholactone (10) is due to the accumulation of the cells in the G2 + M phase of the cell cycle. PMID- 10479314 TI - DNA polymerase beta inhibitors from Sandoricum koetjape. AB - Bioassay-guided fractionation of Sandoricum koetjape using an assay sensitive to DNA polymerase beta inhibition led to the isolation of three active compounds (1 3) having IC(50) values from 20 to 36 microM. Derivatives 5-14 were prepared from compounds 1 and 2; derivatives 11, 12, and 13 showed activity against DNA polymerase beta with IC(50) values ranging from 16 to 36 microM. PMID- 10479315 TI - Six new taxane diterpenoids from the seeds of taxus chinensis var. mairei and taxus yunnanensis AB - Six new taxane diterpenoids and three known 2(3-->20)abeotaxoids were isolated from the seeds of Taxus chinensis var. mairei and of Taxus yunnanensis. Their structures were established as 2alpha, 20-dihydroxy-9alpha-acetoxytaxa-4(20),11 dien-13-one (1), 5alpha-cinnamoyloxy-7beta,10beta, 13alpha-triacetoxy-2(3- >20)abeotaxa-2alpha-ol-4(20),11-dien-9-one (2), 5alpha-cinnamoyloxy-10beta, 13alpha-diacetoxy-2(3-->20)abeotaxa-2alpha,7beta-diol-4(20), 11-dien-9-one (3), 5alpha-cinnamoyloxy-2alpha,7beta,10beta, 13alpha-tetraacetoxy-2(3-->20)abeotaxa 4(20),11-dien-9-one (4), 7-O-acetyltaxine A (5), 2alpha,7beta,10beta, 13alpha tetraacetoxy-5alpha-phenylisoserinatoxy-2(3-->20)abeotaxa-4(2 0),11-dien-9-one (6), 5-O-acetyltaxinine M (7), 5alpha-hydroxy-2alpha,7beta,10beta, 13alpha tetraacetoxy-2(3-->20)abeotaxa-4(20),11-dien-9-one (8), and 2alpha,5alpha dihydroxy-7beta,10beta, 13alpha-triacetoxy-2(3-->20)abeotaxa-4(20),11-dien-9-one (9), on the basis of 1D and 2D NMR spectroscopic analysis. Compound 1, as a new compound isolated from T. chinensis var. mairei, is a rare example of a taxane with a 4,11-diene unit, which is considered to be an important intermediate in the biogenesis of taxoids. Compounds 2-4, 6, and 7 are new compounds, isolated from T. yunnanensis, which also afforded the known compounds 5, 8, and 9. PMID- 10479316 TI - Tryptamine derived amides and acetogenins from the seeds of Rollinia mucosa. AB - Bioactivity-directed fractionation of a CHCl(3)-MeOH (1:1) extract prepared from the seeds of Rollinia mucosa led to the isolation of a mixture of eight novel tryptamine amides. Extensive HPLC allowed the isolation of the major component of the mixture, which was characterized as N-lignoceroyltryptamine (6) using a combination of spectroscopic and chemical methods. The minor amides were identified by GC-MS analysis as N-palmitoyltryptamine (1), N-stearoyltryptamine (2), N-arachidoyltryptamine (3), N-behenoyltryptamine (4), N tricosanoyltryptamine (5), N-pentacosanoyltryptamine (7), and N cerotoyltryptamine (8). Two lignans (pinoresinol dimethyl ether and magnolin) and six acetogenins [membranacin (9), desacetyluvaricin (10), rolliniastatin 1, bullatacin, squamocin, and motrilin] were also isolated. The cytotoxicity of membranacin (9) and desacetyluvaricin (10) against six human solid tumor cell lines was determined. The absolute configuration of the former is reported. PMID- 10479317 TI - Total synthesis of the cytotoxic threo, trans, threo, trans, threo annonaceous acetogenin asimin and its C-10 epimer: unambiguous confirmation of absolute stereochemistry. AB - A convergent synthesis of asimin (1) and its C-10 epimer 33 is reported. The essential features of this synthesis include (a) the addition of an enantioenriched gamma-OMOM allylic indium reagent to a core C-23 aldehyde precursor to install the C-24-C-34 segment with concomitant introduction of the C 24 and C-23 stereocenters; (b) the addition of an enantioenriched gamma-OMOM allylic indium reagent to a core C-16 aldehyde to install the C-10-C-15 segment with formation of the C-15 and C-16 stereocenters, (c) the addition of a dialkyl zinc reagent, catalyzed by a chiral triflamide-Ti(O-i-Pr)(4) complex, to introduce the C-1-C-9 segment with creation of either the 10(R) or 10(S) stereocenters; and (d) aldol condensation of the foregoing C-1-C-34 segment with OTBS-protected lactic aldehyde to incorporate the C-35-C-37 butenolide segment. Removal of the three MOM protecting groups was achieved with aqueous HCl in THF. The 10(R) diastereomer was found to correspond to natural asimin. PMID- 10479318 TI - Triterpenoid saponins from Ilex kudincha. AB - Ten new triterpene saponins, ilekudinosides A-J (2, 6-8, 10, 13-17), together with seven known triterpene saponins, ilexoside XLVIII (1); cynarasaponin C (5); latifolosides A (9), C (3), G (12), and H (4); and kudinoside G (11), were isolated from an aqueous extract of the leaves of Ilex kudincha. They possessed oleanane- and ursane-type triterpenoids as the aglycons. The structures were elucidated by 1D and 2D NMR experiments, including ROE difference, HOHAHA difference, 1H-1H COSY, and 1H-13C COSY (HMQC, HMBC) methods and sugar analysis. Compounds 1 and 5 exhibited acyl CoA cholesteryl acyl transferase (ACAT) inhibitory activity. PMID- 10479319 TI - Potential antipsoriatic agents: lapacho compounds as potent inhibitors of HaCaT cell growth. AB - A number of lapacho compounds, representing the most common constituents of the inner bark of Tabebuia impetiginosa, together with some synthetic analogues, were evaluated in vitro against the growth of the human keratinocyte cell line HaCaT. With an IC(50) value of 0.7 microM, beta-lapachone (4) displayed activity comparable to that of the antipsoriatic drug anthralin. 2-Acetyl-8 hydroxynaphtho[2,3-b]furan-4,9-dione (7), which was prepared in a four-step synthesis from 2,8-dihydroxy-1, 4-naphthoquinone, was the most potent inhibitor among the known lapacho-derived compounds and inhibited cell growth with an IC(50) value of 0.35 microM. Furthermore, other active constituents of lapacho inhibited keratinocyte growth, with IC(50) values in the range of 0.5-3.0 microM. However, as already observed with anthralin, treatment of HaCaT cells with these potent lapacho compounds also caused remarkable damage to the plasma membrane. This was documented by leakage of lactate dehydrogenase into the culture medium, which significantly exceeded that of the vehicle control. Because of their potent activity against the growth of human keratinocytes, some lapacho-derived compounds appear to be promising as effective antipsoriatic agents. PMID- 10479320 TI - New geodiamolides from the sponge Cymbastela sp. collected in Papua New Guinea. AB - Geodiamolides J-P (11-17) and R (19), eight new cyclic depsipetides, have been isolated from the marine sponge Cymbastela sp. collected in Papua New Guinea. The serine residue in geodiamolides L-P (13-17) and R (19) has not been previously found in this family of compounds. PMID- 10479321 TI - Metabolism of paeonol in rats. AB - As a part of our studies on the metabolism of active components from traditional Chinese medicines, paeonol was orally administered to rats. The urinary metabolites were analyzed by 3D HPLC, and their structures were determined to be 2, 4-dihydroxyacetophenone-5-O-sulfate (P1), resacetophenone-2-O-sulfate (P2), 2 hydroxy-4-methoxyacetophenone-5-O-sulfate (P3), paeonol-2-O-sulfate(P4), resacetophenone (P5), and unchanged paeonol, on the basis of their chemical and spectral data. Among these metabolites, P2-P4 and paeonol were detected in the plasma after the oral administration of paeonol. Furthermore, the bile of rats given paeonol orally was found to contain P3, suggesting the enterohepatic circulation of paeonol. PMID- 10479322 TI - Antinociceptive activity of niga-ichigoside F1 from Rubus imperialis. AB - This work describes the antinociceptive effect of a triterpene glycoside, niga ichigoside F1 (1), obtained from an EtOAc extract of the aerial parts of Rubus imperialis. When evaluated against an HOAc-induced writhing model, it exhibited an ID(50) value of 3.1 mg/kg (ip). Moreover, in a formalin-induced pain model, both phases of pain were inhibited by compound 1, with ID(50) values of 2.6 (first phase) and 2.7 (second phase) mg/kg, (ip), respectively. PMID- 10479323 TI - Isolation of two highly methylated polyketide derivatives from a yew-associated penicillium species AB - Two selective antifungal agents were produced by a Penicillium sp. isolated from the inner bark of the Pacific yew tree, Taxus brevifolia. The structures of these highly methylated polyketide derivatives were deduced by detailed analysis of both 1D and 2D NMR and difference NOE spectra. Both compounds were active against the plant pathogen, Sclerotinia sclerotiorum in the standard disk assay. PMID- 10479324 TI - A formylating agent by dehydration of the natural product DIMBOA AB - The natural aglucone 2,4-dihydroxy-7-methoxy-2H-1, 4-benzoxazin-3(4H)-one (DIMBOA, 1) of maize underwent spontaneous dehydration and rearrangement to form 3-formyl-6-methoxybenzoxazolin-2(3H)-one (FMBOA, 2) on reaction with N ethoxycarbonyl-trichloroacetaldimine. Compound 2 was proven to be a reactive formyl donor toward N-, O-, and S-nucleophiles, which may be important in case 2 is formed under biological conditions. PMID- 10479325 TI - A new cyclized 9,11-secosterol enol-ether from the australian sponge euryspongia arenaria AB - The polar fraction of the crude extract from the sponge Euryspongia arenaria was separated by chromatography. Structure elucidation by spectrometric methods allowed the identification of a new steroid, stellattasterenol (2), containing an unprecedented seven-membered cyclic enol-ether in ring C. A related known compound, stellettasterol (3), a pentahydroxy-9,11-secosteroid, was also identified. PMID- 10479327 TI - Isolation and identification of dihydroartemisinic acid hydroperoxide from artemisia annua: A novel biosynthetic precursor of artemisinin AB - Dihydroartemisinic acid hydroperoxide (2) was isolated for the first time as a natural product from the plant Artemisia annua in a 29% yield. Its structure was identified by (1)H and (13)C NMR spectroscopy. Compound 2 is known as an intermediate of the photochemical oxidation of dihydroartemisinic acid (1) leading to artemisinin (3). The presence of 1 and 2 in the plant and the conditions under which 1 can be converted into 2, which can very easily oxidize to 3, provide evidence for a nonenzymatic, photochemical conversion of 1 into 3, in vivo, in the plant. PMID- 10479326 TI - 5-Oxonoraporphines from Mitrephora cf. maingayi. AB - Two new 5-oxonoraporphines, 1 and 2, together with three known compounds, ouregidione, 3-methoxycepharadione B, and isoelemicin, have been isolated from the bark of Mitrephora cf. maingayi. Structures of 1 and 2 were determined to be 1,2,3-trimethoxy-5-oxonoraporphine and 1,2-dimethoxy-3-hydroxy-5-oxonoraporphine on the basis of NMR and MS studies. PMID- 10479328 TI - Neocrotocembranal from Croton oblongifolius. AB - A new cembranoid diterpene, neocrotocembranal (3), was isolated from the stem bark of Croton oblongifolius. Its structure was established on the basis of spectroscopic analysis. This compound inhibited platelet aggregation induced by thrombin, with an IC50 value of 47.21 microg/mL, and exhibited cytotoxicity against P-388 cells in vitro, with an IC(50) value of 6.48 microg/mL. PMID- 10479330 TI - Kahalalide K: A new cyclic depsipeptide from the hawaiian green alga bryopsis species AB - Kahalalide K (1), a new cyclic depsipeptide, was isolated from the Hawaiian green alga Bryopsis sp. Kahalalide K was determined to possess a new array of three L- and three D-amino acids, including a 3-hydroxy-9-methyldecanoic acid that had been previously reported in kahalalides E, H, and J. PMID- 10479329 TI - Phenylethanoid glycosides from Globularia trichosantha. AB - Five phenylethanoid glycosides, crenatoside (= oraposide) (1), verbascoside (= acteoside) (2), trichosanthoside A (3), rossicaside A (4), and trichosanthoside B (5), were isolated from the aerial parts of Globularia trichosantha. Compounds 3 and 5 are new natural compounds, and their structures were established as 3, 4 dihydroxy-beta-phenylethoxy-O-beta-D-xylopyranosyl-(1-->4)-alpha -L- rhamnopyranosyl-(1-->3)-4-O-caffeoyl-beta-D-glucopyranoside and 3, 4-dihydroxy beta-phenylethoxy-O-[beta-D-xylopyranosyl-(1-->4)-alph a-L -rhamnopyranosyl-(1- >3)]-[beta-D-xylopyranosyl-(1-->6)]-4-O-caffeoyl -beta-D-glucopyranoside, respectively. The structures of all compounds were established by spectral evidence. Compounds 1-5 also demonstrated scavenging properties toward the 2, 2 diphenyl-1-picrylhydrazyl radical in TLC autographic assays. PMID- 10479331 TI - A new labdane diterpenoid from Renealmia alpinia collected in the Suriname rainforest. AB - Continuation of a previous study on Renealmia alpinia resulted in the isolation of the new labdane diterpenoid 3, together with two known diterpenoids. The structure of the new diterpenoid was determined by a combination of NMR techniques and HRFABMS. PMID- 10479332 TI - New phenanthrene derivatives from maxillaria densa(1) AB - Two new phenanthrene derivatives, 2,5-dihydroxy-3, 4-dimethoxyphenanthrene (1) and 9,10-dihydro-2,5-dihydroxy-3, 4-dimethoxyphenanthrene (2), were isolated from an extract prepared from the whole plant of the orchid Maxillaria densa with spasmolytic activity. In addition, four known compounds, namely 2,7-dihydroxy-3, 4-dimethoxyphenanthrene, 9,10-dihydro-2,7-dihydroxy-3, 4-dimethoxyphenanthrene (3), 2,5-dihydroxy-3,4, 9-trimethoxyphe-nanthrene, and 2,7-dihydroxy-3,4, 9 trimethoxyphenanthrene, were obtained. The structures of the isolated compounds were elucidated by spectroscopic methods. In the case of phenanthrene derivatives 1 and 3, the structures were unambiguously assigned by X-ray analysis. PMID- 10479333 TI - Two new flavone glycosides from paullinia pinnata AB - Phytochemical investigation of the leaves of Paullinia pinnata L. (Sapindaceae) resulted in the isolation of the two new flavone glycosides characterized as diosmetin-7-O-(2' '-O-beta-D-apiofuranosyl-6' '-acetyl-beta-D-glucopyranoside) (1) and tricetin-4'-O-methyl-7-O-(2' '-O-beta-D-apiofuranosyl-6' '-acetyl-beta-D glucopyranoside) (2). PMID- 10479334 TI - Dichlorodiaportin, diaportinol, and diaportinic acid: three novel isocoumarins from penicillium nalgiovense AB - Three novel isocoumarin (or isochromen) metabolites, dichlorodiaportin [3-(3, 3 dichloro-2-hydroxy-propyl)-8-hydroxy-6-methoxy-isochromen-1-one] (1), diaportinol [3-(2, 3-dihydroxy-propyl)-8-hydroxy-6-methoxy-isochromen-1-one] (2), and diaportinic acid [2-hydroxy-3-(8-hydroxy-6-methoxy-1-oxo-1H-isochromen-3-yl) propanoic acid] (3), were isolated from the cultures of Penicillium nalgiovense along with citreoisocoumarin (4) and 6-methyl-citreoisocoumarin (5). Their structures were elucidated by spectroscopic methods including UV, MS, and NMR. PMID- 10479335 TI - Triterpene trimers from Maytenus scutioides: cycloaddition compounds? AB - Two novel trimers, triscutins A and B (1 and 2), based on pristimerin triterpene units, were isolated and characterized from Maytenus scutioides. Their structures were determined on the basis of spectroscopic evidence, including 1H-13C heteronuclear correlation (HMQC), long-range correlation with inverse detection (HMBC), and ROESY NMR experiments; and their absolute configurations, by means of CD studies. Compounds 1 and 2 were assayed for antimicrobial and cytotoxic activities, and their possible biosynthetic route is proposed. PMID- 10479336 TI - Polyoxygenated flavones from the leaves of comptonella microcarpa AB - The leaves of Comptonella microcarpa have yielded one alkaloid, dictamnine, and four known polyoxygenated flavonoids, meliternatin, 3,5,8-trimethoxy-6,7-3',4' dimethylenedioxyflavone, 7-(3-methylbut-2-enyloxy)-3,5,8-trimethoxy-3', 4' methylenedioxyflavone (3), 7-hydroxy-3,5,8-trimethoxy-3', 4'methylenedioxyflavone. In addition, two new flavonoids were found whose structures were established on the basis of their spectral data as 7-hydroxy 3,5,6,8-tetramethoxy-3',4'-methylenedioxyflavone (1) and 7-(3-methylbut-2 enyloxy)-3,5,6,8-tetramethoxy-3', 4'-methylenedioxyflavone (2). PMID- 10479337 TI - Halisulfate 7, a new sesterterpene sulfate from a sponge, Coscinoderma sp. AB - A new sesterterpene sulfate, halisulfate 7 (1), has been isolated from a Coscinoderma sp. of sponge collected at Yap, Federated States of Micronesia. The structure was determined from spectroscopic evidence. PMID- 10479338 TI - Two new alkaloids from artabotrys uncinatus AB - A novel oxazoloaporphine, artabonatine A (1), and a new 7-hydroxyaporphine, artabonatine B (2), have been isolated and characterized from the fresh unripe fruits of Artabotrys uncinatus, along with five known compounds. Structure elucidation of 1 and 2 was based on UV, IR, NMR, and MS analyses. PMID- 10479339 TI - Three new butenolide lipids from the caribbean gorgonian pterogorgia anceps AB - Three novel fatty acid derivatives (1-3), containing one or two butenolide moieties, were isolated from the Caribbean gorgonian Pterogorgia anceps and chemically characterized by spectroscopic methods and comparison with known compounds. The new molecules were structurally related to ancepsenolide (4), a typical metabolite from Pterogorgia species, which was not detected in this collection of P. anceps. PMID- 10479340 TI - Laxifloranone, a new phloroglucinol derivative from Marila laxiflora. AB - A new polyisoprenylated phloroglucinol derivative has been isolated from the twigs of Marila laxiflora and characterized on the basis of 1D and 2D NMR spectra. Laxifloranone (1) shows moderate inhibition of the cytopathic effects of in vitro HIV infection. PMID- 10479341 TI - A new bislabdane-type diterpenoid from the roots of cunninghamia lanceolata AB - Lanceolatic acid (1), a new bislabdane-type diterpenoid, has been isolated from the roots of Cunninghamia lanceolata. Its structure was elucidated on the basis of spectroscopic data interpretation and confirmed by single-crystal X-ray diffraction analysis. PMID- 10479342 TI - Three new antibacterial alkaloids from a marine sponge Stelletta species. AB - From a marine sponge of the genus Stelletta we isolated three alkaloids, stellettazole B (3), stellettazole C (4), and stellettamide C (5), which were antibacterial against Escherichia coli. Their structures were determined on the basis of spectral and chemical methods. PMID- 10479344 TI - Secobatzellines A and B, two new enzyme inhibitors from a deep-water Caribbean sponge of the genus Batzella. AB - Secobatzelline A (1), a new batzelline natural analogue, and secobatzelline B (2), a likely artifact formed during the isolation procedure, have been isolated from a deep-water marine sponge of the genus Batzella. Secobatzellines A and B inhibited the phosphatase activity of calcineurin, and secobatzelline A inhibited the peptidase activity of CPP32. Both compounds showed in vitro cytotoxicity against P-388 and A-549 cell lines. The isolation and structure elucidation of secobatzellines A (1) and B (2) are described. PMID- 10479343 TI - Homo- and nor-plakotenin, new carboxylic acids from the Palauan sponge Plakortis lita. AB - A specimen of Plakortis lita from Palau yielded the new carboxylic acids, homo plakotenin (2a), the sodium salt of homo-plakotenin (2b), the sodium salt of nor plakotenin (3), the sodium salt of plakotenin (1b), and the known compound plakotenin (1a). The structures of the new acids were elucidated by interpretation of spectral data and by comparison with the known compound. Compounds 1a, 1b, and 2a were found to significantly reduce proliferation of rheumatoid synovial fibroblasts. PMID- 10479345 TI - Unconventional dissolution methodologies. PMID- 10479346 TI - UPPER III: unified physical property estimation relationships. Application to non hydrogen bonding aromatic compounds. AB - The UPPER scheme uses four additive and two nonadditive parameters and several well-known equations to calculate 21 physical properties of organic compounds strictly from molecular structure. The scheme allows reasonable estimations of melting and boiling points, aqueous and octanol solubilities, air-octanol, air water, and octanol-water partition coefficients, vapor pressure, and other properties. In this report non-hydrogen bonding aromatic compounds are used to evaluate a portion of the UPPER scheme. PMID- 10479347 TI - Preparation of a microcrystalline suspension formulation of Lys(B28)Pro(B29) human insulin with ultralente properties. AB - The monomeric analogue, Lys(B28)Pro(B29)-human insulin (LysPro), has been crystallized using similar conditions employed to prepare extended-acting insulin ultralente formulations. In the presence of zinc ions, sodium acetate and sodium chloride, but without phenolic preservative, LysPro surprisingly forms small rhombohedral crystals with similar morphology to human insulin ultralente crystals with a mean particle size of 20 +/- 1 microm. X-ray powder diffraction studies on the LysPro crystals prior to dilution in ultralente vehicle ([NaCl] = 1.2 M) revealed the presence of T(3)R(3)(f) hexamers. Consistent with human insulin ultralente preparations, LysPro crystals formulated as an ultralente suspension ([NaCl] = 0. 12 M) contain T(6) hexamers indicating that a conformational change occurs in the hexamer units of the crystals upon dilution of the salt concentration. The pharmacological properties of subcutaneously administered ultralente LysPro (ULP) were compared to ultralente human insulin (UHI) using a conscious dog model (n = 5) with glucose levels clamped at basal. There were no statistically significant differences between the kinetic and dynamic responses of ULP compared to UHI [C(max) (ng/mL): 3.58 +/- 0.76, ULP and 3.61 +/- 0. 66, UHI; T(max) (min): 226 +/- 30, ULP and 185 +/- 42, UHI; R(max) (mg/kg min): 11.2 +/- 1.9, ULP and 13.3 +/- 2.0, UHI; and T(Rmax) (min): 336 +/- 11, ULP and 285 +/- 57, UHI]. Although the Pro to Lys sequence inversion destabilizes insulin self-assembly and greatly alters the time action of soluble LysPro preparations, this modification has now been found neither to prevent the formation of ultralente crystals in the absence of phenolics nor to compromise the protracted activity of the insulin analogue suspension. PMID- 10479348 TI - The correlation and prediction of the solubility of compounds in water using an amended solvation energy relationship. AB - The aqueous solubility of liquids and solids, as log S(W), has been correlated with an amended solvation equation that incorporates a term in Sigma alpha(2)(H) x Sigma beta(2)(H), where the latter are the hydrogen bond acidity and basicity of the solutes, respectively. Application to a training set of 594 compounds led to a correlation equation with a standard deviation, SD, of 0.56 log units. For a test set of 65 compounds, the SD was 0.50 log units, and for a combined correlation equation for 659 compounds, the SD was 0.56 log units. The correlation equations enable the factors that influence aqueous solubility to be revealed. The hydrogen-bond propensity of a compound always leads to an increase in solubility, even though the Sigma alpha(2)(H) x Sigma beta(2)(H) term opposes solubility due to interactions in the liquid or solid. Increase in solute dipolarity/polarizability increases solubility, whereas an increase in solute excess molar refraction, and especially, volume decrease solubility. The solubility of Bronsted acids and bases is discussed, and corrections for the fraction of neutral species in the saturated solution are graphically presented. PMID- 10479349 TI - Solubility of recombinant human tissue factor pathway inhibitor. AB - Study of recombinant human tissue factor pathway inhibitor (rhTFPI) solubility shows (1) an inverted bell-shaped pH-solubility profile with a broad solubility minimum between pH 5 and 10 such that the solubility minimum midpoint is 2-3 pH units away from its isoelectric point; (2) a negative temperature-solubility coefficient; (3) a strong dependence of solubility on the valence of electrolytes, with both multivalent cations and anions enhancing this effect; and (4) a significant increase of solubility in the presence of charged polymers. At pH 6-7, rhTFPI solubility-salt profiles display typical salting-in and salting out biphasic effects. At a slightly lower pH (pH 5), a third phase in addition to the salting-in and salting-out phases was observed at low ionic strength conditions (5 to 50 mM) where rhTFPI solubility increased as salt concentration decreased. The salting-out constant for rhTFPI in NaCl is 1.04 M(-1) and is independent of the pH of the solution. Resolubilization of rhTFPI precipitates revealed that "insolubility precipitates" (seen during buffer exchanges) resulted from protein solute saturation and could be redissolved by "native" solvent conditions. On the other hand, "instability precipitates" (typically seen after exposure to elevated temperatures or extended storage periods) were caused by insoluble protein aggregate formation and required strongly denaturing conditions to redissolve. PMID- 10479350 TI - Improvement in solubility and dissolution rate of 1, 2-dithiole-3-thiones upon complexation with beta-cyclodextrin and its hydroxypropyl and sulfobutyl ether-7 derivatives. AB - Inclusion complexes between beta-cyclodextrin derivatives and 1, 2-dithione-3 thiones were studied in aqueous solution and in the solid state. Phase solubility study was used to evaluate the complexation in solution, at 37 degrees C, of three cyclodextrins, i. e., beta-cyclodextrin (betaCD), hydroxypropyl-beta cyclodextrin (HPbetaCD), sulfobutyl ether-7-beta-cyclodextrin (SBE7betaCD), and four 1,2-dithiole-3-thiones, i.e., the parent compound dithiolethione (DTT), dimethyldithiolethione (DMDTT), 5-phenyldithiolethione (5PDTT), and anetholetrithione (ATT). Stability constants of the DTT complexes with HPbetaCD and SBE7betaCD were also determined spectrophotometrically using a nonlinear least-squares methodology. Differential scanning calorimetry (DSC) and scanning electronic microscopy (SEM) were used to characterize spray-dried complexes formed between 5PDTT and SBE7betaCD, ATT and SBE7betaCD. Dissolution studies using the USP paddle method were carried out in water at 37 degrees C for both ATT and 5PDTT binary systems with HPbetaCD and SBE7betaCD. Solubility enhancements were much greater with the more lipophilic ATT and 5PDTT compared to DTT and DMDTT, whatever the cyclodextrin used, in the rank order SBE7betaCD > HPbetaCD >> betaCD. Stability constants obtained (between 120 and 12800 mol(-1)) were also the highest for the more lipophilic drugs and in the same rank order SBE7betaCD > HPbetaCD >> betaCD. Results obtained by UV spectrophotometry were in good agreement with those obtained by phase-solubility study. DSC thermograms of spray-dried complexes of ATT and 5PDTT with HPbetaCD and SBE7betaCD lacked the endothermal peak of pure drug peak which was found for the physical mixtures (107 degrees C and 125 degrees C for ATT and 5PDTT, respectively). Finally, dissolution profiles of spray-dried inclusion complexes studied displayed a faster dissolution rate compared to physical mixtures and pure drugs. The present study showed that complexation of 1,2-dithiole-3-thiones with beta-cyclodextrin derivatives resulted in an increase in solubility, allowing intravenous formulation for bioavailability and metabolism studies and an increase in the dissolution rate of the drugs, which should be of interest for oral absorption of these lipophilic compounds. PMID- 10479351 TI - O-ethylphosphatidylcholine: A metabolizable cationic phospholipid which is a serum-compatible DNA transfection agent. AB - 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine was prepared in a one-step reaction from phosphatidylcholine by reaction with ethyl trifluoromethanesulfonate. This and related O-alkyl phosphatidylcholines constitute the first chemically stable triesters of biological lipid structures and the first cationic derivatives of phospholipids consisting entirely of biological metabolites linked with ester bonds. The complex of cationic phospholipid and plasmid DNA transfected cells with high efficiency. Maximum efficiency of transfection was obtained with complexes in which the positive charge was a few percent in excess over the negative charge. Modest stimulation of transfection of common cell lines was obtained by continuous culture in the presence of 10% serum. Incubation of the phospholipid complex for at least 2 h at 37 degrees C in nearly pure serum had no deleterious effects on transfection efficiency. The lipid has low toxicity; BHK cells tolerated amounts of 2 mg/2 x 10(6) cells at concentrations of 1 mg/mL. The lipid is biodegradable; it was hydrolyzed by phospholipase A(2) in vitro and was metabolized with a half-life of a few days in cells in culture. The synthetic route to cationic phospholipids is well suited to the preparation of derivatives that are tailor-made to have a wide variety of different properties. PMID- 10479352 TI - Hydroxypropyl-beta-cyclodextrin inhibits spray-drying-induced inactivation of beta-galactosidase. AB - The single-step, fast spray-drying process may represent a valuable alternative to the multistep, time-consuming freeze-drying process in the area of formulation and processing of biopharmaceuticals. In this study, we tested the use of sucrose and hydroxypropyl-beta-cyclodextrin (HP-beta-CD) as stabilizing excipients in the spray-drying of a model protein, beta-galactosidase. The solutions were processed using a Buchi 190 cocurrent Mini Spray Dryer at an outlet temperature of 61 +/- 2 degrees C. The powders were redissolved and analyzed for catalytic activity, aggregation, chemical decomposition, and thermal susceptibility as observed by high-resolution calorimetry. Spray-drying significantly inactivated beta galactosidase. Spray-drying beta-galactosidase in the presence of sucrose did not prevent inactivation. However, after spray-drying beta-galactosidase in the presence of HP-beta-CD, or HP-beta-CD and sucrose, full catalytic activity was exhibited on reconstitution. Furthermore, the reconstituted product was unchanged in terms of molecular weight, charge, and thermal stability. These findings are consistent with a hypothesis that the change responsible for inactivation of beta galactosidase was mainly a monomolecular, noncovalent change, i. e., the formation of incorrect structures, that arose from surface denaturation. This study clearly demonstrates that cyclodextrins can be useful stabilizing excipients in the preparation of spray-dried protein pharmaceuticals. PMID- 10479353 TI - Biopharmaceutics of boronated radiosensitizers: liposomal formulation of MnBOPP (manganese chelate of 2,4-(alpha, beta-dihydroxyethyl) deuterioporphyrin IX) and comparative toxicity in mice. AB - Binary treatment modalities such as photodynamic therapy (PDT) and neutron capture therapy (NCT) combine low-toxicity electromagnetic irradiation with an appropriate radiation sensitizer to enhance selectivity for tumor targets. The porphyrin derivative tetrakiscarborane carboxylate ester of 2,4-(alpha, beta dihydroxyethyl) deuterioporphyrin IX (BOPP) shows tumor-selective uptake and is active in both treatment modalities. BOPP also chelates paramagnetic ions such as Mn(2+), and therefore its tissue accumulation and selectivity can be detected noninvasively by using magnetic resonance imaging. However, local and systemic toxicity appears elevated for the Mn(2+) chelate (MnBOPP), but is poorly characterized. Here we have developed a liposomal formulation of MnBOPP and compared its toxicity with that of MnBOPP administered to mice in saline. The optimal liposome composition and maximal capacity to accommodate MnBOPP were investigated by differential scanning calorimetry and by encapsulation efficiency. MnBOPP was encapsulated quantitatively at up to 12 mol % (drug:lipid) in liposomes of varying composition, and remained incorporated during extended dialysis. Phase separation of drug- and lipid-rich domains was observed above 12% drug. MnBOPP in buffered saline was lethal to animals at 90 micromol/kg, and caused severe necrosis at the injection site at dose levels of 60 micromol/kg or greater. In contrast, MnBOPP formulated in liposomes was well tolerated at the highest tested dose of 135 micromol/kg, with the elimination of local toxicity. PMID- 10479354 TI - Pharmacokinetics of fluphenazine, a highly lipophilic drug, estimated from a pulse dose of a stable isotopomer in dogs at steady state. AB - The potential utility of a pulse dose of a deuterium-labeled isotopomer (FLU D(4)) in elucidating the pharmacokinetics of fluphenazine (FLU) at steady state was investigated in dogs. The single-dose oral pharmacokinetics of FLU in dogs were established. After resting the dogs for 3 weeks, the animals were dosed to steady state with oral FLU administered at 12-h intervals. Following 15 doses, one dose of FLU was replaced by a pulse dose of FLU-D(4), after which dosing with FLU was resumed. FLU and FLU-D(4) plasma concentrations were determined by tandem mass spectrometry. Comparable estimates of apparent oral clearance were calculated from (i) a single dose of FLU, (ii) a pulse dose of FLU-D(4), and (iii) over a dosing interval at steady state. Average steady-state plasma concentrations were reliably predictable from a pulse dose of FLU-D(4). PMID- 10479355 TI - A novel prodrug approach for tertiary amines. 2. Physicochemical and in vitro enzymatic evaluation of selected N-phosphonooxymethyl prodrugs. AB - Quaternary amine prodrugs resulting from N-phosphonooxymethyl derivatization of the tertiary amine functionality of drugs represents a novel approach for improving their water solubility. Separate reports have demonstrated the synthetic feasibility and rapid and quantitative prodrug to parent drug conversion in rats and dogs. This work is a preliminary evaluation of the physicochemical and in vitro enzymatic reversion properties of selected prodrugs. The loxapine prodrug had over a 15 000-fold increase in aqueous solubility relative to loxapine free base at pH 7.4. The loxapine prodrug was also shown to be quite stable at neutral pH values. The time for degradation product (parent drug) precipitation from an aqueous prodrug formulation would be expected to dictate the shelf life. Using this assumption, together with solubility and elevated temperature chemical stability studies, the shelf life of a parenteral formulation of the loxapine prodrug was projected to be close to 2 years at pH 7.4 and 25 degrees C. In addition, the prodrugs of cinnarizine and loxapine have been shown to be substrates for alkaline phosphatase, an enzyme found throughout the human body, and revert to the parent compound in its presence. The results from these evaluations demonstrate that the derivatives examined have many of the ideal properties required for potential clinical application. PMID- 10479356 TI - A novel prodrug approach for tertiary amines. 3. In vivo evaluation of two N phosphonooxymethyl prodrugs in rats and dogs. AB - N-phosphonooxymethyl derivatives of tertiary amine containing drugs have been identified as a novel prodrug approach for improving aqueous solubility. The in vivo reversion of two prodrugs to the corresponding parent compounds following iv and im administration to rats and dogs was investigated. Equimolar doses of parent drugs (loxapine or cinnarizine) and the corresponding prodrugs were each administered via a rapid iv infusion to rats and dogs. Equimolar doses of loxapine and its prodrug were each administered im to rats only. Blood samples were collected over 12 h, and plasma was assayed for both parent drug and intact prodrug by HPLC. Comparison of the plasma AUC for the parent drugs following administration of the parent drugs and prodrugs allowed estimation of the apparent bioavailability of parent drug from prodrug dosing. Plasma levels of the prodrugs fell below the limit of detection 5 min after iv infusion with an approximate half-life of 1 min. The mean AUCs following iv and im dosing of parent drugs were not statistically different from the parent drug AUCs obtained after prodrug dosing. The results are consistent with rapid and quantitative prodrug to parent drug reversion following administration of the phosphonooxymethyl prodrugs to the rats and dogs. This information, together with previous studies on the synthesis and physicochemical evaluation of the prodrugs, suggests that this novel prodrug strategy is a very promising approach for overcoming solubility limitations seen with many tertiary amine containing drugs at physiological pH values. PMID- 10479357 TI - Oral delivery of insulin using pH-responsive complexation gels. AB - The goal of oral insulin delivery devices is to protect the sensitive drug from proteolytic degradation in the stomach and upper portion of the small intestine. In this work, we investigate the use of pH-responsive, poly(methacrylic-g ethylene glycol) hydrogels as oral delivery vehicles for insulin. Insulin was loaded into polymeric microspheres and administered orally to healthy and diabetic Wistar rats. In the acidic environment of the stomach, the gels were unswollen due to the formation of intermolecular polymer complexes. The insulin remained in the gel and was protected from proteolytic degradation. In the basic and neutral environments of the intestine, the complexes dissociated which resulted in rapid gel swelling and insulin release. Within 2 h of administration of the insulin-containing polymers, strong dose-dependent hypoglycemic effects were observed in both healthy and diabetic rats. These effects lasted for up to 8 h following administration. PMID- 10479358 TI - Assessment of complex peptide degradation pathways via structured multicompartmental modeling approaches: the metabolism of dynorphin A1-13 and related fragments in human plasma. AB - Peptide metabolic pathways in blood or other tissues are often complex because multiple enzyme systems are involved in the degradation of parent drug and its metabolites. Michaelis-Menten-type studies with isolated enzymes have been frequently employed for evaluating the metabolism of peptides. Alternatively, studies with selective enzyme inhibitors or the evaluation of the area under the drug- or metabolite-time profiles have been employed. We tested in this study the usefulness of a multicompartmental pharmacokinetic approach for the assessment of the apparent first-order metabolism of dynorphin A1-13 up to the fourth metabolite generation in human plasma. This multicompartmental kinetic analysis proved instrumental in clarifying ambiguous degradation pathways not easily detectable by the other methods of assessment (enzyme inhibition studies and noncompartmental analysis) because of the lack of specific enzyme inhibitors or specificity problems of the analytical technique employed. The proposed multicompartmental fitting approach was also highly suitable to verify the overall metabolic pathways suggested by the other methods up to the fourth metabolite by testing whether the rate constants obtained by these methods are suitable to describe the overall degradation profile after Dyn A1-13 degradation. Local sensitivity analysis for the degradation of DYNA 1-13 revealed that the model was, however, not able to adequately identify on its own all of the parameters involved in the degradation of dynorphin A1-13. Thus, the method proved beneficial in evaluating and testing the correctness of the overall degradation pathways suggested by other methods. PMID- 10479359 TI - Solubilization of flavopiridol by pH control combined with cosolvents, surfactants, or complexants. AB - This study investigates the roles of both ionized and un-ionized species of flavopiridol in solubilization by complexation, micellization, and cosolvency. Control of pH was used in combination with surfactants (polysorbate 20 and polysorbate 80), cosolvents (ethanol and propylene glycol), as well as uncharged and anionic complexing agents [hydroxypropyl beta-cyclodextrin (HPbetaCD) and sulfobutyl ether beta-cyclodextrin (SBEbetaCD)] to solubilize flavopiridol. These combined techniques increase not only the solubility of the un-ionized flavopiridol but also the solubility of the ionized drug. This study confirms that previously developed equations effectively characterize the roles of pH, pK(a), and either complexation constant, micelle partition coefficient, or cosolvent solubilizing power in determining drug total aqueous solubility. PMID- 10479360 TI - Microfabricated microneedles: A novel approach to transdermal drug delivery PMID- 10479361 TI - A critical review of methodology and interpretation of mirror self-recognition research in nonhuman primates. AB - In this paper we critically review conceptual and methodological issues of mirror self-exploration research. We conclude that: (1) mirror self-exploration provides evidence for mirror self-recognition; (2) inter- and intraspecies differences in terms of presence or absence of mirror self-recognition are not yet sufficiently clear, that is, whether its absence in species and individuals is methodological (false negative with respect to trait) or ontological (true negative with respect to trait) in nature; and (3) proposals of hypotheses to account phylogenetically for the existence of inter- and intraspecies differences in mirror self recognition are premature. Before hypotheses such as these are advanced, the impact of rearing conditions on mirror self-recognition should be examined further. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479362 TI - Fin-flicking behaviour: a visual antipredator alarm signal in a characin fish, Hemigrammus erythrozonus. AB - Glowlight tetras, Hemigrammus erythrozonus, show a conspicuous visual display ('fin flicking') upon detecting a conspecific chemical alarm signal (alarm pheromone). We conducted laboratory studies to test two, nonmutually exclusive hypotheses regarding the function of this behaviour. To test the hypothesis that fin flicking acts as a visual alarm signal to conspecifics, we paired shoals of tetras with a stimulus tetra in an adjacent tank (separated by a one-way mirror) and exposed the stimulus tetras to conspecific skin extract (with alarm pheromone) or swordtail skin extract (which lacks ostariophysan alarm pheromone). Stimulus tetras significantly increased their frequency of fin flicking when exposed to conspecific skin extract, but not to the swordtail skin extract control. The test shoals showed a significant increase in antipredator behaviour in response to fin-flicking stimulus tetras, but not to stimulus tetras that did not fin-flick. To test the hypothesis that fin flicking serves as a predator deterrent, we paired predatory Jack Dempsey cichlids, Cichlasoma octofaciatum, with tetras in adjacent tanks and exposed the tetras to either conspecific or swordtail skin extract. Predators remained further away from the stimulus tetra, took longer to initiate a bite and displayed fewer bites towards tetras that fin flicked compared with tetras that did not fin-flick. These results support our hypothesis that fin flicking in tetras serves as both a visual alarm signal to conspecifics and a deterring signal to predators, and suggest that individual tetras may reduce their risk of predation by fin flicking upon detecting a predation threat. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479363 TI - The effect of proximity on landmark use in Clark's nutcrackers. AB - Clark's nutcrackers, Nucifraga columbiana, store thousands of pine seeds during the autumn and retrieve them throughout the winter. It has been shown that these birds are able to use visual cues to relocate hidden food in the laboratory. In this set of experiments, we trained three groups of Clark's nutcrackers to find a hidden food goal that was placed in the same spatial location relative to the testing room. During training, the location of two local cues in relation to the goal differed between the three groups. Group 1 learned the task with the cues closest to the goal, group 2 with the cues further from the goal, and group 3 with the cues furthest from the goal. To test whether the proximity of these two local cues to the goal affected how spatial information was used, we manipulated local and global information in a series of experiments. Results indicated that local cues were more important sources of spatial information for group 1, whereas global cues seemed to be used more by birds in groups 2 and 3. These findings suggest that the proximity of visual cues lead to overshadowing of other sources of spatial information surrounding the goal. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479364 TI - Illumination and the perception of remote habitat patches by white-footed mice. AB - Perceptual range, or the distance at which habitat 'patches' can be perceived, constrains an animal's informational window on a given landscape. If such constraints are great, they may limit successful dispersal between distant habitat patches. On dark nights, nocturnal white-footed mice, Peromyscus leucopus, have surprisingly limited perceptual abilities regarding distant forested habitat. In fact, their ability to orient towards such habitat while travelling in a bare agricultural field indicates a perceptual range under 30 m. However, increasing illumination can increase perceptual range. For example, full moonlight extends the perceptual range of mice to about 60 m. Light levels at dusk (twilight) extend perceptual range still further to about 90 m. These results suggest that interpatch dispersal by white-footed mice would be more successful under greater illumination, but travelling under such conditions entails a considerable risk of predation. These mice might avoid such a conflict by travelling under the cover of darkness with the aid of information gathered remotely during relatively high illumination. We show that mice are indeed capable of such a 'look now and move later' strategy: mice retain directional information gained under bright conditions and maintain a previously determined bearing in conditions under which distant navigational stimuli may be largely absent (e.g. maximal darkness). Ultimately, a better understanding of the behavioural and ecological factors affecting the movements of animals across landscapes should produce a clearer picture of the interaction between landscape structure and population ecology. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479365 TI - Social determinants of reproductive failure in male common marmosets housed with their natal family. AB - Postpubertal male common marmosets, Callithrix jacchus, engage in little or no sexual behaviour while living with their natal families. The social mechanisms underlying this phenomenon have not been identified but have been assumed to include reproductive suppression by dominant males and/or avoidance of mating with closely related females. We evaluated these two possible components of male reproductive failure. Seven postpubertal males and their fathers underwent a series of 45-min sex tests, in which the son, the father, or the son and father together were allowed to interact freely with the mother/mate or with an unfamiliar, unrelated adult female. We measured testosterone, luteinizing hormone and cortisol concentrations in blood samples collected from males immediately following each test, and in basal blood samples collected on three different occasions. Sons, but not fathers, engaged in very low rates of sexual behaviour when tested with the mother/mate. When tested with unrelated females, however, sons engaged in significantly more sexual behaviour than they had when tested with their mothers and showed no differences from their fathers. When sons and fathers were tested together, both males performed significantly less sexual behaviour than when tested alone with a female; however, they showed no overt competition for females and engaged in little agonism. Hormone levels did not differ significantly between fathers and sons. These results indicate that avoidance of mating with familiar females and possibly father-son competition, but not specific, rank-related suppression, contribute to reproductive failure among postpubertal male marmosets living with their natal family. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479366 TI - The effects of hunger on locomotory behaviour in two species of wolf spider (Araneae, Lycosidae). AB - We compared the influence of recent feeding history on locomotory behaviour in two species of wolf spiders, Hogna helluo (Walckenaer) and Pardosa milvina Hentz, in the laboratory. Both species are cursorial hunters. We maintained the spiders in the laboratory on satiation and stringent feeding regimes and measured their locomotory activity levels for 1 h using a digital activity recording device. We subjected H. helluo to either ad libitum feeding for 14 days or no food for 14 days. We subjected P. milvina to ad libitum feeding or fasting treatments for 7 and 14 days. We found that H. helluo showed a shift in locomotory activity depending on feeding regime, whereas P. milvina did not. Food-limited H. helluo travelled further than satiated H. helluo, and did so by moving more frequently. Pardosa milvina was in general much more active than H. helluo. We propose that P. milvina is an active forager compared with the sit-and-wait strategy of H. helluo. This difference in foraging strategy is correlated with differences in body size and habitat use. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479367 TI - Honest signalling during prey-predator interactions in the lizard Anolis cristatellus. AB - Current theory on the evolution of pursuit-deterrent signals predicts that it may be advantageous for the prey to communicate to the predator its alertness and its ability to escape an attack. I tested these predictions by staging predator-prey encounters between A. cristatellus lizards and a model of one of its predators under natural conditions. Results supported the use of pushup displays as pursuit deterrent signals. The intensity of signals, measured as the number of pushups given during predation episodes, was significantly positively correlated with individual physiological condition measured as endurance capacity. Because endurance capacity can be a critical aspect limiting the ability of A. cristatellus to escape a predatory attack, pushup displays can potentially communicate an individual's ability to escape an attack and, therefore, can be categorized as honest signals. Furthermore, because pushup displays are widely used during anoline social interactions, predation pressure and sexual selection may simultaneously favour the evolution of honest communication to allow both the predator and the potential mate or male rival to assess individual quality using the same signal. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479368 TI - Proximate factors associated with high levels of extraconsort fertilization in polygynous grey seals. AB - Behavioural estimates of male mating success in polygynous grey seals, Halichoerus grypus, may be misleading as females are known to be promiscuous. At Sable Island, Nova Scotia, we collected behavioural observations and skin samples for paternity analysis from 56 females and their attending males. Twenty-four of these females were found in the following year and their offspring were sampled. Using seven hypervariable microsatellite loci, we excluded the consort male as the father in 43% of the cases. The probability of exclusion of these seven loci was 98.2%. Contrary to expectations, inland females had higher rates of extraconsort fertilizations (ECFs) (70%) than beach females (23%). Younger females (<9 years) had slightly more ECFs than older females, but this was not significant. The duration of male consortship did not differ between females with ECFs and females fertilized by their consort male. Two explanations may account for the inland females having more ECFs: a higher ratio of females to tenured males inland may provide a greater opportunity for nonconsort males to obtain copulations; and inland females travel greater distances to depart for the ocean and may attract more males. These results are more consistent with the hypothesis that ECFs are a by-product for females of male strategies to maximize reproductive success than with hypotheses concerned with either material or genetic benefits gained by females. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479369 TI - Back to the basics of antipredatory vigilance: can nonvigilant animals detect attack? AB - Many birds and mammals respond to a heightened risk of predation, especially that associated with smaller group sizes, with an increase in vigilance. All interpretations of the way in which vigilance responds to changes in predation risk assume that animals feeding with their heads down (i.e. animals in a nonvigilant state) cannot detect approaching predators. We provide the first explicit test of this assumption by 'flying' a mounted hawk down a 15-m chute towards actively feeding, free-living, dark-eyed juncos, Junco hyemalis. Juncos were targeted individually for simulated attack when they had either a 'head down' view up the chute, or a completely unobstructed view; a junco with a head down view could see up the chute only when it lowered its head to feed. Juncos with an unobstructed view almost always detected the hawk at the maximum distance of 15 m. Juncos with a head-down view usually detected the attack at a distance of 10-15 m against a grey background, but detection distances were shorter when attacks occurred against a camouflaged background. The results demonstrate that these birds have a considerable ability to detect approaching predators even when not overtly vigilant, although their detection ability is greater when they raise their heads. Vigilance sequences, therefore, probably consist of bouts of low quality detection (active feeding) interspersed with bouts of higher-quality detection (overt vigilance) that can only be accomplished at the expense of feeding. This realization has major implications for current interpretations of the vigilance group size effect and antipredator vigilance in general. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479370 TI - Siblicide revisited in the spotted hyaena: does it conform to obligate or facultative models? AB - Aggression among spotted hyaena, Crocuta crocuta, siblings is often intense, and sometimes lethal. Frank et al. (1991, Science, 252, 702-704) proposed that siblicide routinely occurs in half of all spotted hyaena litters, namely those composed of same-sex twins. We propose an alternative to this 'obligate' model. In our 'facultative' model we suggest that siblicide is far less common than previously supposed, and that it occurs only when resources are insufficient to sustain two cubs. According to this facultative model, intense neonatal aggression functions to establish intralitter dominance rather than to kill siblings. Furthermore, differences in litter size and composition between captive and field settings previously used to support the obligate model are assumed in the facultative model to be due to prenatal factors rather than to postnatal siblicide. Here we tested the predictions of these two hypotheses with 10 years of field data from hyaenas inhabiting the Masai Mara National Reserve, Kenya. We found that, although sex compositions of hyaena litters averaged over the entire 10-year period did not differ from those predicted by chance, they did vary with environmental conditions. Litter sizes, by contrast, remained constant. These data are inconsistent with the widely accepted hypothesis that spotted hyaenas in same-sex litters routinely engage in siblicide. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479371 TI - Feather mites, pectoral muscle condition, wing length and plumage coloration of passerines. AB - I compared the feather mite (Acari, Proctophyllodidae) loads of moulting birds with features of the new plumage that they were growing. I examined 21 samples, each sample containing individuals of the same species, sex and age class (juvenile, yearling or adult). I used nine species: wren, Troglodytes troglodytes; dunnock, Prunella modularis; robin, Erithacus rubecula; blue tit, Parus caeruleus; great tit, P. major; chaffinch, Fringilla coelebs; greenfinch, Carduelis chloris; linnet, C. cannabina; and yellowhammer, Emberiza citrinella. As previously reported for the house finch, Carpodacus mexicanus, birds with more feather mites grew duller plumage and relatively shorter wings than less infested individuals of the same sex and age class. They also had lower protein reserves judged by the shape of their pectoral muscle. Mite load was usually a better predictor of coloration than pectoral muscle score, but the opposite was true for changes in wing length. In contrast to house finches, birds whose plumage was dull before moult had higher mite loads while moulting. Even if feather mites do not cause dull plumage or short wings, they appear to be a good indicator of birds in poor physiological condition. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479372 TI - Housing and welfare in laboratory rats: effects of cage stocking density and behavioural predictors of welfare. AB - Using male and female Alderley Park (Wistar-derived) rats housed in single-sex groups in standard laboratory cages, we looked at the effect of group size (one, three, five or eight) on competitive behaviour and time budgeting (initial and longer term), changes in their serum testosterone (males), corticosterone and antibody concentrations, and organ pathology at age 16 weeks, together with the interrelationships between behavioural measures and pathophysiological indices of social stress. Group size had only limited long-term effects on overall time budgeting and did not affect pathophysiological responses, although there were highly significant differences between individuals in replicate cage groups. Pathophysiology within both sexes showed strong and highly specific correlations with a small subset of behaviours suggesting frustrated attempts to escape from cages, including chewing the cage bars. Escape-related behaviour also correlated strongly with one component of competitive behaviour, Aggressive Grooming within both sexes, although Aggressive Grooming correlated with pathophysiological responses only among males. Females generally showed greater escape-related behaviour associated with greater signs of pathophysiology regardless of the level of aggression shown between cagemates. Major differences in intercorrelated behavioural and pathophysiological responses between replicate groups implied that the individual composition of groups rather than their size had the greater impact on the welfare of the rats, especially among females. This may be consistent with adaptive sex differences in their competitive reproductive strategies. The frequency of apparent escape-related behaviours and Aggressive Grooming, particularly when rats are first introduced into their cage groups, may provide a simple assessment of the welfare implications of particular cage groupings. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479373 TI - Effect of tagging marine turtles on nesting behaviour and reproductive success. AB - We studied green, Chelonia mydas, and loggerhead, Caretta caretta, turtles nesting on the island of Cyprus to determine the effects of flipper tagging on postovipositional behaviour and reproductive success. Tagging was undertaken immediately after oviposition (64 green turtles; 111 loggerheads). On 12 occasions, loggerheads immediately stopped covering the eggs and proceeded directly to the sea. No green turtles abandoned nesting. In all other cases, where sufficient data were collected, the duration of the two postovipositional phases of nesting behaviour (covering and camouflaging) and the speed of descent to the sea did not differ between tagged and untagged turtles. The durations of behaviours also did not differ between females not tagged, tagged once or tagged twice. There was no effect of tagging on the likelihood of hatching or on the hatching success of clutches in either species. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479374 TI - Effect of experience of occlusion events on the domestic chick's strategy for locating a concealed imprinting object. AB - We investigated how positional and directional cues affected the way single domestic chicks, Gallus gallus domesticus, searched for a concealed imprinting object. The chicks were 4 or 12 days of age and had been reared in enriched pens that allowed them to move out of sight of the imprinting object, or control pens that did not. In experiment 1, 44 chicks viewed a positional cue, which was a stationary imprinting object placed for 1 min in front of one of two possible hiding locations (each concealing an identical imprinting object) either to the left or right of the chick. When released, chicks moved towards, and spent more time in, the side in which the positional cue had been presented. We presented a further 44 chicks with a directional cue, which involved an imprinting object moving from near one possible hiding location towards the other and its occlusion midway between the two possible hiding locations. When released, chicks moved towards, and spent more time in, the opposite side to that indicated by the visible direction of movement of the imprinting object. Additionally, chicks reared in enriched pens spent more time in the side in which the imprinting object was seen than chicks reared in control pens. We examined the relative influence of positional and directional cues in experiment 2, by providing both cues sequentially to 88 chicks. Chicks spent more time in the positional cue side, when the directional cue offered complementary information, but no side preference was found when cues provided contradictory information. We conclude that in this study, orientation towards a concealed object was based on previous spatial location alone, with little influence from the temporal aspects of the presentation such as movement before occlusion or last-seen location. We discuss the possibility that chicks imprinting on a stationary object develop a strategy for locating a concealed social stimulus that is unrepresentative of chicks reared by a hen. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479375 TI - Algorithmic behaviour and spatial memory are used by two intertidal fish species to solve the radial maze. AB - We used an eight-arm radial maze to assess the relative contributions of learned patterns of movement (algorithmic behaviour) and spatial memory to the foraging efficiency of two sympatric rocky-shore fish, fifteen-spined stickleback, Spinachia spinachia, and corkwing wrasse, Crenilabrus melops, exploiting nonrenewable food sources. To forage efficiently, subjects had to avoid arms already depleted within a trial. In the absence of spatial cues, sticklebacks and wrasse improved their foraging efficiency by developing the algorithm of visiting every third arm. In the presence of spatial cues (coloured tiles) algorithmic behaviour was largely subsumed by the use of spatial memory. Imposition of a delay within trials reset the behavioural algorithm, so depressing foraging efficiency in the absence of cues, but not in their presence when memory could be used to guide behaviour. Memory retention for previous choices (working memory) lay within the range 0.5-5.0 min, consistent with the characteristic timescale expected for habitats where prey distribution changes rapidly during the tidal cycle. We considered two hypotheses on the type of information memorized: the cue list hypothesis and the spatial configuration hypothesis. The cue list hypothesis predicts that neither random repositioning nor fixed rotation of spatial cues should impair foraging efficiency, whereas the spatial configuration hypothesis predicts that efficiency should be impaired by random repositioning of cues but not by rotation. Data supported the spatial configuration hypothesis. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479376 TI - Is infestation the result of adaptive choice behaviour by the parasite? A study of mites and midges. AB - We studied the possibility that a parasite, a hydracharinid mite, chooses its host, a chironomid midge. Mites, Unionicola ypsilophora, were placed into containers with either one or two Chironomus plumosus pupae and we counted the number of mites on the midges after the latter had emerged. Fewer mites were rejected by hosts when they had a choice of host, suggesting that they actively choose which individual to parasitize. In several midge species collected in the wild, fluctuating asymmetry in wing length was a good predictor of mite infestation. We suggest that mites use some correlate of fluctuating asymmetry to make their choice of host. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479377 TI - Preimaginal conditioning in Drosophila revisited. AB - During metamorphosis, the nervous system of a holometabolous insect changes significantly. Attempts to demonstrate preimaginal conditioning, here taken to mean the retention of learning through metamorphosis, have given mixed results. We used two behavioural assays (the T maze and trap assay) to see whether a change in adult responsiveness could be induced by exposing Drosophila melanogaster larvae to a conditioning stimulus. There was no evidence for preimaginal conditioning from either assay, but the trap assay demonstrated that menthol contamination from the larval environment on the puparial surface could induce a change in adult behaviour. Exposure of adult insects to this contamination could give the appearance of preimaginal conditioning, when in fact the behavioural induction occurred during the adult stage. Young flies responded less strongly than older flies to the odour cues in both assays. This may explain the apparently contradictory findings of some earlier studies of preimaginal conditioning. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479378 TI - The effects of environmental context on laboratory rat social recognition. AB - Moving an animal from the environmental context in which it has learned a particular task to an entirely different context can reduce performance. We investigated the effect of switching environmental contexts on the ability of adult laboratory rats, Rattus norvegicus, to recognize and habituate to repeated presentations of juvenile conspecifics. Adults were exposed to juveniles for four periods of 5 min, separated by a 15-min interval. Rats either received all four exposures in the same context, or the first three in one context and the fourth in a different context. Half the rats in this latter group were familiarized with both contexts prior to testing, the other half had no experience of either. In all groups, the adults reduced their investigation of the juveniles over the three initial exposures. Mild aggression increased over the same period for the context-unfamiliar rats. A significant reduction in investigation by these rats between the third and fourth exposures, when the context was changed, suggested that the context switch further increased habituation to the juveniles. However, the context-familiar rats showed no such change, indicating that the changes observed for the context-unfamiliar rats were due to the effect of context novelty. This was supported by the finding that, during the first exposure, context-familiar adults investigated juveniles more and were more aggressive than those for which the contexts were novel. These results suggest that familiar contextual cues play only a minor role in the short-term social memory of laboratory rats. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479379 TI - Optimal distances for squirrels to transport and hoard walnuts. AB - Walnuts, Juglans airanthifolia, are an important food item for Japanese squirrels, Sciurus lis, which hoard them for later use. We set up feeders in small walnut patches in secondary forest, mounted radiotransmitters on nuts, placed them on the feeders and tracked them when they were cached. Of 720 nuts placed on the feeders over a 3-year period, 32.6% were eaten immediately and 50.6% were scatter-hoarded 0-168 m from the feeders. With increasing distance to the cache site, the squirrels took longer to return to the feeders (range 0.35-35 min). However, the further away a cache was, the less likely it was to be stolen by conspecifics or other nut-eating animals: nuts we hoarded ourselves disappeared more quickly from near the source trees. Squirrels took nuts further away later in the season when caches were lost at a higher rate. Using a simple model maximizing the rate of energy gain per unit time taken to search for and hoard the nuts at different distances from the source, we discuss whether squirrels alter the transport distance of hoarding nuts according to cache survival rate and/or nut availability. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479380 TI - Recognition of conspecifics by honeybee guards uses nonheritable cues acquired in the adult stage. AB - Social insects use both environmental and heritable (genetic) recognition cues when discriminating between nestmates and non-nestmates. In honeybees, Apis mellifera, both types of recognition cues are used, although their relative importance depends upon context, experimental design and environmental factors. Our aim in this study was to investigate which cues honeybees use to discriminate between nestmates and non-nestmates under natural conditions. To do this, we used an assay of nestmate recognition that very closely simulated natural conditions, with naturally occurring guards at actual nest entrances and workers that had been naturally reared and had spent their adult life in a hive. Guards exclusively used environmental recognition cues when encountering entering bees. There was no evidence that guards used heritable cues: related nestmates and unrelated nestmates, unrelated non-nestmates and related non-nestmates all had the same probability of being accepted (ca. 80 versus 20%). Possible reasons why heritable cues are not used are discussed. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479381 TI - Drosophila song as a species-specific mating signal and the behavioural importance of Kyriacou & Hall cycles in D. melanogaster song. AB - 'Species-specific' mating signals may not make a significant contribution to sexual isolation if differences between species are not matched by narrowly tuned differences in female preferences. The courtship song of the melanogaster species group of Drosophila has been studied by several researchers as a potential factor in sexual isolation, but there are few clear demonstrations that species differences are behaviourally significant. We played synthetic song with typical pulse interval parameters for D. melanogaster, D. simulans or D. sechellia to D. melanogaster or D. simulans females in the presence of mute males. Females mated most quickly when stimulated by song typical of their own species, confirming that song can influence sexual isolation between these species. This effect was greater in D. melanogaster than D. simulans, which correlates with reduced variability in mean interpulse interval within D. melanogaster. We also examined the role of 'Kyriacou & Hall cycles' (periodic cycling of mean interpulse intervals) in D. melanogaster. Contrary to some predictions, the presence of the Kyriacou & Hall cycle was more important in increasing mating speed than the increased variability of interpulse intervals that is inevitably present in song containing a cycle. The large stimulatory effect of cycles on mating speed is curious, given that the periodicity of cycles would probably be difficult for females to detect during courtship in field conditions. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479382 TI - The ontogenetic switch between odonate life history stages: effects on fitness when time and food are limited. AB - During the course of ontogeny, odonates switch from being aquatic larvae to being terrestrial adults. Ontogenetic niche shift theory proposes that such shifts are adaptive and have evolved to maximize a growth rate (size) to mortality rate ratio. Individuals should therefore switch from one niche to the other at an optimal size or state. Since the majority of odonates are seasonal breeders, the extent to which the switch is optimal will depend upon the time and the resources available during postembryonic development. We collected a cohort of larvae that varied in how close they were to eclosion and reared them on either a high nutrition or a low-nutrition diet. We then determined the relative influence of both time and nutritional constraints on survival and development rate, as well as the body size, size-corrected flight muscle mass and fat reserves of individuals at eclosion. Damselflies in both high- and low-nutrition treatments responded to a short development period by developing faster and reducing their body size, but did not change their proportional investment in fat reserves and flight muscle. Reduced larval nutrition resulted in decreased body size, flight muscle mass and fat reserves at eclosion. However, it had no effect on survival to eclosion, or development rate. We discuss these results in terms of the influence that time and nutritional constraints have on odonate development patterns and fitness. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479383 TI - Female preference for multiple partners: sperm competition in the hide beetle, Dermestes maculatus (DeGeer). AB - Explanations for the evolution of polyandry emphasize either direct benefits or indirect genetic benefits. The former class of explanation predicts that females may seek multiple copulations but not necessarily multiple partners, while the latter predicts that females will seek multiple copulations with different partners. We investigated these predictions experimentally in the hide beetle, in which females mate more than once during their reproductive cycle. Female hide beetles remated more readily with novel males than with their previous partners, suggesting they mate multiply in order to obtain indirect, genetic benefits. In double-mating experiments with sterile males, the proportion of eggs fertilized by the second male was highly variable; this could not be explained by variation in male body size, duration of copulation or mating order. Male hide beetles form postinsemination associations by riding on the backs of females. This behaviour was more prevalent in the presence of rival males and less prevalent in the presence of additional females, suggesting that the behaviour serves to increase paternity, perhaps by obtaining additional copulations. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479384 TI - Competitive foraging in the social spider Stegodyphus dumicola. AB - Reproductive skew models have greatly enhanced the study of sociality but are applicable only to societies where the aim of the members of the group is to reproduce. In 'foraging societies', where the aim of the members of the group is to grow, quite different parameters will govern the form that the societies take. We examined factors that influence the structure of foraging groups in the social spider Stegodyphus dumicola (Eresidae). In particular, we examined food distribution and consumption within groups, and how this related to predictions derived from two models. One model indicates that colonies in which individuals compete for resources via interference competition are more likely to survive than those in which individuals divide resources using scramble competition. The second model predicts the proportions of hunters, cheaters and scavengers expected in foraging groups. We found that food was not distributed evenly among group members, and that spiders that fed primarily on the head and thorax of the prey during the middle of a feeding event gained the most body mass. Spiders even lost mass if they fed only in the last hour of a foraging event. Large spiders had a competitive advantage (via interference competition) in obtaining preferred positions, and could ingest food faster than small spiders. Distributing food among colony members in this manner could cause large size differences between colony members, as predicted by the models. The implications of competitive foraging for sociality are discussed. Copyright 1999 The Association for the Study of Animal Behaviour. PMID- 10479386 TI - Probing the immunological properties of the extracellular domains of the human beta(1)-adrenoceptor. AB - The human beta(1)-adrenoceptor is an immune target for autoantibodies with functional activity in cardiovascular diseases. Different epitopes on the extracellular domains of the receptor are involved. To study the immunological and pharmacological properties of these epitopes, rabbits were immunized with peptides corresponding to a large domain in the N-terminal part of the receptor and to its first and second extracellular loops. In contrast to the two other peptides, the first extracellular loop did not have immunogenic properties but acted as a hapten. Antibodies affinity-purified with the three synthetic peptides were able to significantly immunoprecipitate the solubilized receptor, confirming that they recognized the target receptor. While antibodies against the N-terminal domain did not inhibit the binding of a radiolabelled antagonist to the receptor, those against the first and second extracellular loop showed non-competitive inhibition. Similarly, only the two latter antibodies exerted a specific agonist like effect on the receptor, as assessed on neonatal rat cardiomyocytes in culture. Our results are in accordance with those found for human anti-receptor autoantibodies with functional effects. We conclude that not all extracellular epitopes give rise to functional autoantibodies with potential physiopathological relevance in cardiac diseases with an autoimmune component. PMID- 10479387 TI - Amelioration of experimental allergic neuritis by sodium fusidate (fusidin): suppression of IFN-gamma and TNF-alpha and enhancement of IL-10. AB - The immunomodulating antibiotic drug fusidic acid and its sodium salt sodium fusidate (fusidin) ameliorate several organ-specific immunoinflammatory diseases. Because preliminary observations suggest that fusidin may also exert a beneficial effect in Guillain-Barre syndrome (GBS), here we have studied the effects of fusidin on actively induced experimental autoimmune neuritis (EAN) in rats, a known animal model for GBS. Both prophylactic and therapeutic treatment with fusidin (4 mg/rat day ip) markedly ameliorated the clinical course of the disease compared to vehicle-treated animals. The beneficial effects were associated with profound modifications of the capacity of these rats to produce and release pro- and anti-inflammatory cytokines such as IFN-gamma, TNF-alpha and IL-10, which are important in regulating the development of EAN. PMID- 10479388 TI - Inhibition of experimental melanin protein-induced uveitis (EMIU) by targeting nitric oxide via phosphatidylcholine-specific phospholipase C. AB - Experimental melanin protein-induced uveitis (EMIU) is an autoimmune uveitis induced by immunization with uveal melanin protein. Fas and FasL enhancement is reported in rats with EMIU. Tricyclodecan-9-yl-xanthogenate (D609), a specific inhibitor of phosphatidylcholine-specific phospholipase C, inhibits inducible nitric oxide synthase (iNOS) induction. In two independent experiments, 35 Lewis rats with EMIU received either D609 or PBS daily. The eyes and draining lymph nodes were collected for histology, analyses of nitrite, peroxide, and superoxide dismutase, Fas and FasL immunochemistry, in situ hybridization for iNOS mRNA and in situ apoptosis detection at the peak of the disease. Both experiments showed significant inhibition of EMIU by D609. Decreases in nitrite and peroxide, increase of superoxide dismutase and lower expressions of iNOS mRNA were found in D609-treated, as compared to PBS-treated eyes. There was mild enhancement of Fas and FasL in the eyes and lymph nodes of D609-injected animals. DNA fragmentation was increased in the lymph nodes of D609-treated rats. We conclude that iNOS activation is responsible for NO production in eyes with EMIU. The suppressive effect of D609 on EMIU may result from scavenging NO and activating apoptosis previously inhibited by NO along with other anti-inflammatory effects. PMID- 10479389 TI - Comparison of immune responses to extracellular domains of mouse and human thyrotropin receptor. AB - The mouse and human thyrotropin receptors show greater than 87% homology in their amino acid sequences. However, glycosylated extracellular domains of mouse (mET gp) and human (hET-gp) thyrotropin receptors showed differences in their ability to react with patient autoantibodies to thyrotropin receptor (TSHR). To test for potential differences in their immunogenicity, we immunized BALB/c mice with either gel pure non-glycosylated ectodomain of human TSHR (ETSHR II), or hET-gp (hET-gp III), or mET-gp (mET-gp III). Alternatively, mice were primed with gel pure hET-gp or mET-gp and subsequently immunized with insect cells expressing hET gp (hET-gp II) or mET-gp (mET-gp II) respectively. All groups of mice immunized with TSHR developed high titers of antibodies against the respective immunogens. As shown earlier, sera obtained from mice immunized with ETSHR showed strong reactivity to peptide 1 (aa 22-41) and weak reactivity to peptides 23 (aa 352 371), 24 (aa 367-386), 25 (aa 382-401), and 26 (aa 397-415). Mice immunized with hET-gp or mET-gp showed comparable titers to peptides 1 and 23 and lower reactivity to other peptides. Mice immunized with hET-gp showed higher TBII reactivity (52.2%) compared to mice immunized with either ETSHR (20.9%) or mET-gp (34.5%). Peptides from the C-terminal region of ETSHR could neutralize the TBII activities of sera from mice immunized with ETSHR or hET-gp but not mET-gp. Compared to corresponding control mice, T4 levels in mET-gp II mice were only marginally higher. These data suggested that outcome of immunization with mouse ETSHR is comparable to that seen after immunization with human ETSHR. PMID- 10479390 TI - Isolation and characterization of two human monoclonal anti-phospholipid IgG from patients with autoimmune disease. AB - The antigenic specificity of anti-phospholipid antibodies (APA) is a matter of intensive investigation. To further characterize these antibodies, we attempted to isolate human monoclonal APA. B-cells of patients with at least one positive test for antibodies against cardiolipin, phosphatidylserine, beta2-glycoprotein I (beta2-GPI) or the lupus anti-coagulant were immortalized by transformation with Epstein-Barr virus and screened for production of specific IgG. Positive pools were fused with a heteromyeloma cell line and APA-secreting clones were isolated by standard procedures. Two monoclonal APA, HL-5B from a 51-year-old man with primary anti-phospholipid syndrome and recurrent cerebral microinfarctions, and RR-7F from a 48-year-old women with systemic lupus erythematosus but no evidence for thrombotic events were obtained. HL-5B is of the IgG2 subtype with lambda light chains, while RR-7F is IgG2 with kappa light chains. Both monoclonals show reactivity against cardiolipin and phosphatidylserine but lack reactivity against beta2-GPI or lupus anti-coagulant activity. To yield the same OD in the cardiolipin and phosphatidylserine ELISAs RR-7F must be used in an approximately 10-fold higher concentration than HL-5B, indicating a lower affinity towards these antigens. Interestingly, both mAPA can bind to cardiolipin in the absence of beta2-GPI. They do not cross-react with dsDNA but show reactivity against oxidized low-density lipoproteins. Analysis of the heavy chain mRNA of HL-5B and RR-7F showed that both are members of the VH3 family. While HL-5B shows extensive somatic mutations in the CDR1 and 2 regions, indicating that it was derived by a T cell-dependent antigen driven process, RR-7F is apparently germline encoded. The two monoclonal APA can be used as tools in further structural and functional analyses. PMID- 10479391 TI - Clinical features and IgG subclass distribution of anti-p80 coilin antibodies. AB - We examined the clinical features of patients presenting antinuclear autoantibodies against p80-coilin and the IgG subclass distribution of anti- p80 coilin antibodies. Sera from 365 Japanese patients were analysed. Immunoblotting and indirect immunofluorescence microscopy techniques were used with a polyclonal rabbit antiserum against p80-coilin. Eleven patients with anti-p80-coilin antibodies were found. All the patients were female and nine were in their twenties. None could be diagnosed with differentiated rheumatic disease except for one case of systemic scleroderma and another of Sjogren's syndrome. Most patients had general fatigue, arthralgia, headaches, dysmenorrhea, lymph node swelling and/or low grade fever such as chronic fatigue syndrome (CFS), and showed low complement. One patient fulfilled the criteria for CFS. All were younger females than those often diagnosed with rheumatic disease in previous reports. Patients' sera had a predominant distribution of subclass IgG(1)anti-p80 coilin antibodies and five sera had concomitant subclass IgG(2). Two rheumatic disease patients had a relatively high titer of IgG(2)anti-p80-coilin antibodies. The IgG(2)subclass of anti-p80-coilin antibodies may be a specific marker for systemic autoimmune disease. PMID- 10479392 TI - Cellular and humoral immune responses against autoreactive T cells in multiple sclerosis patients after T cell vaccination. AB - Myelin basic protein (MBP)-reactive T cells may play an important role in the autoimmune pathogenesis of multiple sclerosis (MS). MBP-reactive T cells can be specifically targeted by T cell vaccination, a procedure whereby MS patients are immunized with attenuated autologous MBP reactive T cells. T cell vaccination induces immune responses to the vaccine cells together with a depletion of MBP reactive T cells. Forty-nine MS patients were treated with T cell vaccination in an extended phase I trial to study the safety, immune responses and clinical effects of T cell vaccination. In the present paper the immune responses towards the vaccine cells were characterized. Substantial long-term in vitro proliferative responses were observed in all treated patients. Some patients, immunized with different clones, displayed distinct proliferative reactivity against the various vaccine clones, suggesting unequal immunogenic properties of these clones. Reactive TCRalphabeta(+), CD8(+)and CD4(+)T cells, and to a lesser extent, gammadelta T cells and NK cells were observed to in vitro stimulation with the vaccine cells. A small fraction only of CD8(+)T cells expressed cytolytic and inhibitory anti-clonotypic reactivity against the vaccine cells. Stimulation with the vaccine clones predominantly induced expression of pro inflammatory cytokines in these mixed cultures, although one vaccine clone consistently induced production of IL-4. CD4(+)T cells are the major cytokine producing cells in these anti-vaccine lines. We could not detect upregulated antibody responses to the vaccine cells in most patients, although a temporary antibody response was observed in one patient. In conclusion, immunization with attenuated autoreactive T cells induces a complex cellular response specifically targeted at the vaccine cells, but no antibody responses. These data provide further insights into the mechanisms of T cell vaccination and improve our understanding of the complex regulatory networks of autoreactive T cells. PMID- 10479393 TI - Constitutive impaired TCR/CD3-mediated activation of T cells in IDDM patients co exist with normal co-stimulation pathways. AB - IDDM is a T cell-mediated autoimmune disease which is paradoxically associated with T cell functional deficiencies. The proliferative response of PBMC under CD3 , Vbeta2-, Vbeta8- and Vbeta7-stimulation was investigated in IDDM and NIDDM patients, non-diabetic first-degree relatives and control subjects. Despite normal surface expression of the TCR/CD3 complex, the TCR/CD3-mediated proliferation of PBMC from IDDM patients was significantly impaired compared to control subjects (P<0.05). This defect was specific for the autoimmune disease, constitutive and not linked to the class II MHC genotype, to metabolic disturbances or to presence of specific autoantibodies. Inefficient activation of T cells was not related to a lower capacity of CD28 to transduce co-stimulative signals because proliferative responses under CD2/CD28 stimulations were similar in IDDM and control groups. The IL-2/IL-2 receptor system was functional because unstimulated PBMC proliferated in response to increasing amounts of IL-2. Nevertheless, despite normal expression of CD25, addition of IL-2 did not normalize the proliferative defect linked to IDDM. In conclusion, excluding a faulty co-stimulation pathway, these results are in favour of a constitutive defect in the CD3/TCR transduction machinery, increasing sensitivity to apoptosis or anergy in T cells from IDDM patients. PMID- 10479394 TI - Autoantibodies to multiple islet autoantigens in patients with abrupt onset type 1 diabetes and diabetes diagnosed with urinary glucose screening. AB - It has been reported that there is a heterogeneity in the clinical course of Japanese patients with type 1 diabetes. To elucidate the associations of expression of autoantibodies to multiple islet antigens with age of onset and mode of diagnosis of diabetes in Japanese patients with type 1 diabetes, autoantibodies against the protein tyrosine phosphatase-like molecules ICA512 (IA 2) and phogrin (IA-2beta) (ICA512/phogrin-A), GAD (GADA), insulin (IAA), and islet cell cytoplasm (ICA) were determined in sera from 73 Japanese patients with type 1 diabetes obtained within 14 days of diagnosis. Patients were divided into groups based on the age of onset (10 years, n=49) or the mode of onset (abrupt onset, n=59 and urinary screening identified, n=14). Of 73 new-onset patients with type 1 diabetes, 43 (59%) and 32 (44%) had ICA512A and phogrin-A levels exceeding the 99th percentile of 184 normal control subjects, respectively. Forty-five patients (62%) were positive for either ICA512A or phogrin-A. The frequencies for other autoantibodies were 71% for GADA, 48% for IAA, and 62% for ICA. The frequency of ICA512/phogrin-A was significantly higher in patients with an age of onset less than 10 years (83%) than in patients aged >10 years (51%, P<0.01). The positivity of ICA512/phogrin-A was less in patients whose diabetes was diagnosed by the urine glucose screening test (21%, P<0.001) than in abrupt onset patients (71%). Combined analysis (>/=1 antibody) of GADA, IAA, and ICA512/phogrin-A detected 88% of abrupt onset and 93% of screening positive patients vs. 70% and 29%, respectively, for ICA (P<0.0005). These results indicate that the expression of ICA512/phogrin-A and cytoplasmic ICA is less in patients identified by urinary glucose testing but indicate that with combined autoantibody testing 90% of patients can be identified independent of the mode of diagnosis. PMID- 10479395 TI - Autoreactive T cell responses in insulin-dependent (Type 1) diabetes mellitus. Report of the first international workshop for standardization of T cell assays. AB - Type 1 diabetes is thought to result from a T cell-mediated destruction of the pancreatic beta-cells. Multiple and sometimes conflicting studies have identified a variety of aberrations in the cellular immune response to autoantigens in persons with the disease. Potential explanations for these discrepancies include incomparable techniques or culture conditions, diversity in the populations of patients or controls tested, and differences in autoantigen preparations. A T cell workshop was organized by the Immunology of Diabetes Society with the aim of appreciating and identifying problems associated with autoreactive T cell assays in type 1 diabetes. As a first phase, a series of candidate autoantigens were analysed by reference laboratories for quality. Subsequently, these preparations, as well as control stimuli, were distributed in a blind fashion to 26 laboratories worldwide, including all experienced centres, for analysis of T cell proliferation assays in 10 recent onset type 1 diabetes and 10 non-diabetic controls. For this analysis, participants used their own assays and references. The islet autoantigen quality control analyses performed prior to the distribution indicate that the quality of recombinant autoantigen preparations requires improvement. For example, several T cell clones specific for glutamic acid decarboxylase (GAD65) were unable to cross-react with GAD65 expressed in baculovirus, yeast or bacteria. Moreover, autoantigens expressed in E. coli interfered with autoantigen-specific proliferation of both T cell clones and peripheral blood mononuclear cells. Nonetheless, responses could be measured to all autoantigen preparations evaluated in the workshop. During the blind phase of the study, all centres were able to reproducibly measure T cell responses to two identical samples of tetanus toxoid, but there was significant interlaboratory variation in sensitivity and extent of the proliferative response measured. Third, the results using candidate autoantigens indicated that although a few laboratories could distinguish type 1 diabetes patients from non-diabetic controls in proliferative responses to individual islet autoantigens, in general, no differences in T cell proliferation between the two groups could be identified. This first T cell workshop on T cell autoreactivity in type 1 diabetes confirms that this was a difficult area for interlaboratory investigations, but provided insight towards future efforts focused on standardizing autoreactive T cell measurements. Some previously reported conflicting results can in part be explained by the observed interlaboratory variability. The inability to discriminate normal controls from new onset type 1 diabetes patients suggests that measuring proliferative responses in PBMC represents an incomplete picture of the immune response, perhaps complicated by difficulties in identifying suitable antigens and assays for standardized use. PMID- 10479396 TI - Developmental sequences in class reasoning and propositional reasoning. AB - This study explored the theoretical prediction that class and propositional reasoning skills emerge as a function of the developing ability to coordinate increasingly complex negation and affirmation operations. Children from Grades 1, 3, 5, and 7 (7-, 9-, 11-, and 13-year-olds) were presented with problems from each domain. Rasch analyses of the children's responses were consistent with the hypothesis that both types of problems measured a single underlying dimension (i.e., the coordination of affirmation and negation operations). Qualitatively distinct levels of class and propositional reasoning were identified along this dimension, adding support to the notion that children's reasoning follows a logical developmental sequence. Planned comparisons supported the order theoretical prediction that different groups of items account for solution differences between grade levels. Results also indicated that children encounter significant difficulties when they have to reason on the basis of negative information. PMID- 10479397 TI - Culture-general and culture-specific factors in the discrimination of melodies. AB - We examined effects of a culture-general factor, pattern redundancy (number of repeated tones), on the discrimination of 5-tone melodies that differed in their adherence to Western tonal conventions. Experiment 1 evaluated the ability of 9 month-old infants to differentiate "standard" melodies from subtly altered "comparison" melodies. Greater redundancy of the standard melodies was associated with enhanced infant performance, but musical conventionality had no effect. Experiment 2 evaluated comparable abilities in 5-year-old children and musically untrained adults. Children's performance was enhanced by the redundancy of standard melodies, but the effect was greater in conventional than in unconventional contexts. The redundancy of standard melodies facilitated adults' performance in conventional but not in unconventional contexts. Thus, increasing musical exposure seems to attenuate the effects of culture-general factors such as pattern redundancy while amplifying the influence of culture-specific factors. PMID- 10479398 TI - Preschoolers' and adults' reliance on object shape and object function for lexical extension. AB - We investigated the developmental progression of reliance on object function versus object shape to extend novel words. In 3 experiments, 3-year-olds, 5-year olds, and adults were presented with sets of objects consisting of a target, a same-shape/different-function match, a different-shape/same-function match, and a distracter. In Experiments 1 and 2, function was emphasized during the word learning phase and participants were given direct experience with the functions of target and test objects. In Experiment 3, function was emphasized both during the learning phase and when requesting a referent of the novel labels. Across all 3 experiments, 3- and 5-year-olds focused on shape while adults focused on function when extending the novel words. These results suggest a developmental change in the consideration of shape and function in lexical extension. PMID- 10479399 TI - Presence of IL-1- and TNF-like molecules in Galleria mellonella (Lepidoptera) haemocytes and in an insect cell line Fromestigmene acraea (Lepidoptera). AB - In this study the authors give immunocytochemical evidence for the presence of interleukin (IL)-1alpha- and tumour necrosis factor (TNF) alpha-like molecules in the haemocytes of last instar larvae from the greater wax moth Galleria mellonella. Similar results are demonstrated in a continuous haemocyte line (BTI EA-1174-A) from the salt marsh caterpillar Estigmene acraea. In Galleria mellonella larvae granular cells show a strong positive reaction with both primary antibodies, whereas plasmatocytes are stained to a lesser extent. Cell line haemocytes also react positively with both antibodies. After activating the cells with lipopolysaccharide (LPS) staining of Estigmene acraea cells is decreased, whereas Galleria mellonella haemocytes show no visible reaction in comparison to non-activated cells. PMID- 10479401 TI - Regulatory elements of the leukaemia inhibitory factor (LIF) promoter in murine bone marrow stromal cells. AB - Leukaemia inhibitory factor (LIF) plays an important role as a haematopoietically active cytokine. As described earlier in a murine model, interleukin 1 (IL-1) induced LIF mRNA and protein expression. We utilized the murine cell line +/+ 1.LDA11 to further define regulatory mechanisms of LIF expression in bone marrow stromal cells. The production of LIF mRNA is stimulated by IL-1beta, TNF-alpha, and the cAMP analogue 8-bromoadenosine 3':5'-monophosphate (8BrcAMP). LIF mRNA expression is controlled at the transcriptional level. Different fragments from 542 to -45 bp 5' upstream of the transcriptional start site of the murine LIF gene were fused to the luciferase gene. All LIF-promoter luciferase constructs exhibited constitutive luciferase activity under serum free conditions. The level of luciferase activity decreased with LIF-promoter constructs of less than 249 bp (pLIF249) in size. When tested with the 314 bp LIF-promoter construct, incubation of stromal cells with IL-1beta (500 U/ml) resulted in a 1.57-fold stimulation, with TNF-alpha (500 U/ml) in 2.06-fold stimulation, and with 8BrcAMP (0.5 mM) in a 3. 42-fold stimulation of luciferase activity. By testing different deletion mutants we could narrow the IL-1 and TNF-alpha responsive promoter areas to the region -249 to -145 bp and the 8BrcAMP responsive area from -145 to -82 bp. Mobility shift experiments revealed that nuclear proteins from stromal cells form a DNA-protein complex by binding to the region from -249 to -145 bp of the LIF promoter. PMID- 10479400 TI - Distinct roles for p42/p44 and p38 mitogen-activated protein kinases in the induction of IL-2 by IL-1. AB - Interleukin 1 (IL-1) activates p42/p44 and p38 mitogen-activated protein kinases (MAP kinases) in target cells. Here we have used two specific inhibitors, PD98059 which inhibits MAP kinase kinase (MEK), and SB203580 which inhibits p38 MAP kinase to explore the involvement of these kinases in the induction of IL-2 by IL 1 in the murine thymoma cell line EL4.NOB-1. Both kinase inhibitors suppressed IL 1-stimulated IL-2 production. PD98059 blocked IL-2 mRNA accumulation and the induction of a reporter gene linked to the IL-2 promoter. In contrast, SB203580 only marginally inhibited IL-2 promoter-linked reporter gene expression and had no inhibitory effect on IL-2 mRNA levels. Neither PD98059 nor SB203580 had an inhibitory effect on NFkappaB-driven reporter gene expression in response to IL 1. Surprisingly, higher concentrations of SB203580 (30 microM) potentiated the IL 1 responses. PD98059 also inhibited induction of IL-2 by phorbol 12-myristate 13 acetate (PMA), and AP1-linked reporter gene expression in response to PMA but not IL-1. These results indicate that p42/p44 MAP kinase is involved in the regulation of IL-2 gene transcription by IL-1, whilst p38 MAP kinase has a post transcriptional target. Additional IL-1 signalling pathways can clearly compensate for the lack of p38 MAP kinase which result in potentiation of the IL 1 responses observed at high-dose SB203580. PMID- 10479402 TI - Regulation of APO-2 ligand/trail expression in NK cells-involvement in NK cell mediated cytotoxicity. AB - Apo-2L is a new member of the tumour necrosis factor (TNF) family shown to induce apoptosis in a number of tumour cell lines. Apo-2L mRNA is expressed by numerous human tissues. Here we report that Apo-2L is expressed and utilized by human Natural Killer (NK) cells. NK cells were shown to express surface Apo-2L in response to interleukin 2 (IL-2) activation, and this response was restricted to the CD3(-)population of the NK cells. Apo-2L mRNA and intracellular Apo-2L were present in both CD3(-)and CD3(+)NK cells; however, increased expression in response to IL-2 was only observed in CD3(-)CD56(+)cells. Also, IL-2-activated NK cells were shown to utilize membrane-bound Apo-2L in mediating lysis of Jurkat cells. Furthermore, Apo-2L-induced apoptosis of Jurkat cells was more rapid than FasL-induced apoptosis, indicating an important and distinct role for Apo-2L in apoptotic cell destruction. In conclusion, we report that NK cells express Apo-2L and that IL-2 activated CD3(-)NK cells utilize the Apo-2L pathway in mediating target cell lysis. PMID- 10479403 TI - Growth inhibition of human promonocytic leukaemic U937 cells by interferon gamma is irreversible and not cell cycle phase-specific. AB - Growth of human promonocytic leukaemic U937 cells was found arrested within 24 h upon exposure to interferon gamma (IFN-gamma). Removal of the interferon did not result in the resumption of growth, as is evident from the absence of doubling of viable cell count and(3)H-thymidine incorporation. 5-Bromo-2'-deoxyuridine-based flow cytometric analysis of the growth-arrested cells, 24 h subsequent to the removal of IFN-gamma, showed absence of DNA synthesis, confirming the irreversible nature of the growth inhibition. Propidium iodide-based flow cytometric analysis of the growth-arrested cells showed a distribution which is typical of a growth inhibition without resulting in the accumulation of cells in any specific phase of the cell cycle. These results indicated that IFN-gamma arrested growth of U937 cells in an irreversible and cell cycle phase-independent manner. These observations were in contrast to our earlier report on the reversible and cell cycle phase-specific growth inhibition of human amniotic (fetal epithelial) WISH cells by the interferon. PMID- 10479404 TI - Modulation of haematopoietic progenitor development by FLT-3 ligand. AB - The Flt-3 receptor is expressed in primitive haematopoietic cells and its ligand exerts proliferative effects on these cells in vitro in synergy with other cytokines. To increase our knowledge of the functional properties of the human Flt-3 ligand (FL) as relating to in vitro expansion of haematopoietic stem cells, the effects on murine haematopoiesis of FL alone or in combination with other growth factors were studied. Analysis of Flk-2/Flt-3 mRNA expression indicated that Flk-2/Flt-3 was preferentially expressed in primitive haematopoietic cell populations. To examine the expression of the Flk-2/Flt-3 receptor on megakaryocyte progenitors (CFU-Meg), Flk-2/Flt-3 positive and negative CD34(+)populations were separated from human bone marrow and cultured in a plasma clot culture system. CFU-Meg colonies were found in the Flk-2/Flt-3 negative fraction. Myeloid (CFU-GM) derived colonies appeared in the presence of FL alone. Neither FL+IL-3 nor FL+IL-3+IL-6 had any effect on the generation of megakaryocyte colonies (CFU-MK), due to the lack of FL receptor expression on megakaryocyte progenitors. Bone marrow cells remaining after 5-fluorouracil (5 FU) treatment of mice represent a very primitive population of progenitors enriched for reconstituting stem cells. This cell population expressed FL receptors, as revealed by RT-PCR analysis. Addition of FL alone did not enhance the replication of such cells in liquid cultures as compared to controls. However, a significantly greater generation of myeloid progenitors (CFU-GM) in clonogenic assays was observed in the presence of FL+IL-3, FL+GM-CSF or FL+CSF-1. In addition, the effects of FL on in vitro expansion of murine haematopoietic stem cells were studied using lineage-negative (lin(-)) Sca-1 positive (Sca-1(+)) c-kit positive (c-kit(+)) marrow cells from 5-FU treated mice. FL enhanced the survival of primitive murine lin(-)Sca-1(+)c-kit(+)cells. FL and IL-6 were able to significantly expand murine progenitor stem cells in vitro and promote their survival. These studies strongly suggest that FL significantly and selectively enhanced the generation of myeloid progenitors in vitro and increased myeloid progenitor responsiveness to later acting growth factors. In addition, FL synergized with IL-6 to support in vitro expansion of haematopoietic progenitors and promoted the survival of lin(-)Sca-1(+)c-kit(+)cells. PMID- 10479405 TI - In vitro modulation of cisplatin resistance by cytokines. AB - We have previously demonstrated that treatment of wild-type (wt) T98G malignant glioma cells with Cisplatin (CDDP) led to a resistant phenotype. It has been demonstrated that interleukin 1 (IL-1) potentiates the cytotoxic effect of CDDP and that IL-6 decreases cytotoxicity by inhibition of apoptosis in cancer cells. Here we examined the influence of IL-1 and IL-6 on the sensitivity of resistant and wt T98G cells. Using semi-quantitative PCR reactions in three independent experiments, resistant glioma cells revealed a decreased IL-1alpha (50.3+/-7.2), IL-1beta (56.0+/-4.0) and IL-6 (44. 3+/-18.2) mRNA content compared to wt cells (100%;P<0.05). Resistant and wt cells were positive for the receptors IL-1RI and IL-6R (PCR). To investigate whether IL-1alpha, IL-1beta or IL-6 changes the sensitivity of the resistant and wt cells towards CDDP, cells were incubated up to 7 days with 10(-5) M CDDP and with different concentrations (0, 0.01, 0.1, 1 ng/ml) of cytokine. Sensitivity was tested in a colorimetric assay (MTT). IL-6 did not influence the sensitivity towards CDDP of either wt or resistant cells, while IL-1alpha and IL-1beta enhanced sensitivity of resistant cells to CDDP. These data suggest that autocrine IL-1 production is involved in the mechanisms of resistance in T98G cells. PMID- 10479406 TI - Differentially increased IL-6 mRNA expression in liver and spleen following injection of liposome-encapsulated haemoglobin. AB - Injection of the red cell substitute liposome-encapsulated haemoglobin (LEH) induces increased serum interleukin (IL)-6 in the absence of other inflammatory cytokines. In vitro studies found that IL-6 mRNA was increased in Mphi and endothelial cell lines by co-culture with LEH. In the present study, cytokine mRNA expression in extracts of livers, spleens, lungs and kidneys after LEH injection was determined by semi-quantitative RT-PCR. The distribution of cells expressing IL-6 mRNA in livers and spleens was visualized by in situ hydridization; extracts of kidney and lung did not show increased IL-6 mRNA and were not studied further. IL-6 mRNA accumulation in livers and spleens was increased at 4 h following LEH injection and had declined by 24 h. In the liver, cells expressing IL-6 mRNA were located in endothelia of hepatic and portal veins, and hepatic sinuses, Kupffer cells and epithelial cells of bile ducts. Endothelium of hepatic arteries did not express IL-6 mRNA. Lymphocytes, haematopoietic cells and macrophages expressed IL-6 mRNA in spleens. The data suggest that cells of the reticuloendothelial system (RES) might be a significant source of increased plasma IL-6 in vivo after LEH administration. PMID- 10479407 TI - Cxc chemokine receptor expression on human endothelial cells. AB - CXC chemokines play a important role in the process of leukocyte recruitment and activation at sites of inflammation. However, recent evidence suggests that these molecules can also regulate endothelial cell functions such as migration, angiogenesis and proliferation. In this study we have investigated CXC chemokine receptor expression in both primary cultures of human umbilical vein endothelial cells (HUVEC) and the spontaneously transformed HUVEC cell line, ECV304. We found that both cell types express mRNA for chemokine receptors CXCR1, CXCR2 and CXCR4, but not CXCR3. Flow cytometric analysis revealed low levels of CXCR1 but higher levels of CXCR4 cell surface expression. HUVECs responded to SDF-1alpha with a rapid and robust calcium flux, however no calcium flux was seen with either IL-8 or Gro-alpha. HUVECs and ECV304 cells did not proliferate in response to CXC chemokines, although ECV304 cells did migrate towards SDF-1alpha and IL-8. These data demonstrate that HUVECs and the endothelial cell line, ECV304 express functional CXC chemokine receptors. PMID- 10479408 TI - Interleukin 10 mitigates the development of the zymosan-induced multiple organ dysfunction syndrome in mice. AB - We investigated the effect of interleukin 10 on the development of zymosan induced multiple organ dysfunction syndrome (MODS) and on plasma concentrations and production capacity of tumour necrosis factor (TNF)-alpha by peritoneal cells. Groups of C57BL/6 mice received a single intraperitoneal injection with zymosan, a cell wall component of Saccharomyces cerevisiae, at day 0. Daily doses of human recombinant interleukin 10 (IL-10: 10 or 50 microg/kg) were given intraperitoneally either starting directly before administration of zymosan (day 0), or 5 or 8 days after administration of zymosan. The animals were monitored for survival, condition, body weight and temperature. On day 12 all surviving animals were killed to obtain plasma, organs and peritoneal cells. Plasma concentrations of TNF-alpha and lipopolysaccharide-stimulated production of TNF alpha by peritoneal cells were measured; organ weights were registered as an indicator for organ damage. IL-10 improves survival and clinical condition and also reduces organ damage, but only at the highest dose used and only when started simultaneously with the administration of zymosan. Circulating TNF-alpha concentrations 12 days after zymosan are not affected by any of the IL-10 schedules used. However, lipopolysaccharide-stimulated production of TNF-alpha by peritoneal cells is increased, in a dose- and time-dependent fashion. The anti inflammatory cytokine IL-10 is able to attenuate the development of MODS in this model, but only when given simultaneously with zymosan, and in high dosages. PMID- 10479409 TI - Immunohistochemical comparative analysis of transforming growth factor alpha, epidermal growth factor, and epidermal growth factor receptor in normal, hyperplastic and neoplastic human prostates. AB - Immunoreaction to TGF-alpha was limited to the basal epithelial cells of focal areas in the normal prostates. In benign prostatic hyperplasia (BPH) the immunostained areas were more widespread and immunolabelling was observed in both basal and columnar (secretory) cells of the epithelium. Some cells in the connective tissue stroma were also stained. In prostatic adenocarcinoma, epithelial immunostaining was even more extensive and intense than in BPH, and some stromal cells were also stained. Epidermal growth factor (EGF) immunostaining was only present in some basal cells in normal prostates. In BPH, this immunoreaction was strong in the basal cells and even stronger in the secretory cells. In prostatic cancer, the intensity of epithelial cell immunoreactivity was intermediate between that of normal prostates and that of BPH specimens. EGF-receptor immunostaining was focal and located in the basal cells in normal prostates. In BPH, labelling was also localized in basal cells but extended to wider areas. Some stromal cells appeared weakly labelled. In the prostatic carcinoma, both basal and columnar cells appeared stained and the number of immunolabelled stromal cells was higher than in BPH. The results presented suggest that, in normal conditions, EGF and TGF-alpha act as autocrine growth factors for the basal cells of the prostatic epithelium. In BPH this action is maintained and, in addition, the columnar cells start to secrete both factors which are bound by the basal cell receptors, giving rise to a paracrine regulation which probably overstimulates basal cell proliferation. In prostatic carcinoma, besides these regulatory mechanisms, the acquisition of EGF-receptors by the secretory cells develops an autocrine regulation which might induce their proliferation. PMID- 10479410 TI - Nuclear targeting of mutant Huntingtin increases toxicity. AB - Huntington's disease is a neurodegenerative disorder resulting from expansion of the polyglutamine region in huntingtin. Although huntingtin is normally cytoplasmic, in affected brain regions proteolytic fragments of mutant huntingtin containing the polyglutamine repeat form intranuclear inclusions. Here, we examine the contribution of nuclear localization to toxicity by transiently transfecting neuro-2a cells with an N-terminal huntingtin fragment similar in size to that believed to be present in patients. The huntingtin fragment, HD-N63, was targeted either to the cytoplasm with a nuclear export signal (NES) or to the nucleus with a nuclear localization signal (NLS). The NES decreased the number of cells with aggregates in the nucleus while an NLS had the opposite effect. By cotransfecting HD-N63 with GFP as a marker, we observed direct cell loss with constructs containing expanded polyglutamine repeats. Compared to unmodified HD N63-75Q, adding an NES reduced cell loss by 57% while an NLS increased cell loss by 111%. These results indicate that nuclear localization of mutant huntingtin fragments plays an important role in cell toxicity. PMID- 10479412 TI - A Thermodynamic Study of Interactions of Monoionic Kaolinite with N,N Dimethylacetamide (DMA) and Pyridine (PY). AB - Two series of monoionic (M) kaolinites (M = H(+), Ca(2+), Cu(2+), Zn(2+), Al(3+), and Fe(3+)) were prepared from a natural Brazilian clay (Sao Simao clay, SP): series A, dried under vacuum at 25 degrees C, and series B, dried under vacuum at 25 degrees C. The natural clay was characterized by elemental analysis, cation exchange capacity, X-ray diffractometry, differential thermal analysis, and surface area measurements (BET). The interactions of M,M-dimethylacetamide and pyridine with these (M) kaolinites were studied by adsorption isotherms, titration calorimetry, and infrared spectroscopy (for pyridine only). The results indicated a coordinative interaction between M,M-dimethylacetamide and (M) kaolinites. For the interactions between pyridine and (M) kaolinites, the formation of hydrogen bonds (M = Al(3+), Fe(3+)), hydrogen and coordinative bonds (M = Cu(2+), Zn(2+)), and ionic pairs (M = H(+), Ca(2+)) was observed. For the pyridine-(Cu) kaolinite system, a substitution of water coordinated to Cu(2+) by pyridine was established. PMID- 10479411 TI - Aberrant presenilin-1 expression downregulates LDL receptor-related protein (LRP): is LRP central to Alzheimer's disease pathogenesis? AB - Low density lipoprotein receptor-related protein (LRP) polymorphisms have recently been associated with an increased susceptibility of Alzheimer's disease (AD). Furthermore, LRP has been linked to molecules that confer susceptibility to AD (apolipoprotein E, alpha-2-macroglobulin, amyloid precursor protein), previously with the exception of the presenilins. Here we report that aberrant presenilin-1 expression in vivo and in vitro downregulates LRP. Specifically, transgenic mice overexpressing the M146L or L286V presenilin-1 mutation show decreased levels of LRP expression in neuronal populations where presenilin-1 and LRP are closely colocalized or coexpressed. Moreover, cell lines transfected with presenilin-1 also expressed decreased levels of LRP. These findings suggest that LRP may be central to AD pathogenesis since all proteins genetically associated with AD can now be linked via a single pathway to LRP. PMID- 10479413 TI - The Turbidity of Cell Nuclei in Suspension: A Complex Case of Light Scattering. AB - The turbidity of a suspension of cell nuclei isolated from animal tissue homogenates is a complex case of non-Rayleigh scattering. As a first approximation to this system, we have characterized a number of factors that may contribute to the observed turbidity: cation-dependent chromatin condensation, thermal denaturation of chromatin, nuclear shrinking, and changes in the optical properties of the membrane bilayer. Small differences in cation concentration, particularly in the case of divalent cations, lead to large changes in chromatin supramolecular organization, thus to large turbidity effects; thermally-induced changes in turbidity have a similar origin, although they are less pronounced. Under certain circumstances, either salts or heat may induce condensation of chromatin, the latter being connected to the inner side of the nuclear envelope, nuclear shrinking ensues, and this in turn modifies the suspension turbidity. Finally, changes in the physical properties of the lipid bilayers or of the phospholipids in the nuclear envelopes may also have significant effects, though smaller than chromatin changes, in the overall turbidity of the nuclear suspension. PMID- 10479414 TI - Capillarity Effects on Viscous Gravity Spreadings of Wetting Liquids. AB - Highly viscous silicone oils within a wide range of volumes were left to spread on smooth horizontal substrates to investigate the effects of capillarity on viscous gravity spreadings under the condition of complete wetting. The study was centered on the intermediate asymptotic behavior, where the details of the initial liquid distribution are irrelevant. We detected small but appreciable departures from the well-known solution without surface tension forces (viscous gravity self similar solution, or base solution). Two stages are clearly identified in the spreadings. During the first, which is usually rather brief, capillarity does not play an appreciable role on the dynamics of the spreading, i.e., the base solution is a very good approximation. The head of the current displays a wheel-like profile progressively decreasing in size; when the size becomes on the order of the capillary length, this stage ends. The wheel-like configuration cannot be associated either with the rheological behavior of the fluid used or with the initial conditions. To observe this first stage without the influences due to peculiarities of the initial conditions, a proper release of the spreading is needed. The second stage is characterized by a spreading rate below the base solution; the slowing is associated with a change of the current head shape, which takes the form of a wedge as a consequence of capillarity. The rate of advance of the front may still be well approximated by the same power law as given by the base solution, but with a smaller prefactor. In this work we measure the parameters that characterize the flow during both stages; besides, for the second stage, we develop a heuristic calculation which shows that the wedge-like shape of the current head gives place to a higher viscous dissipation rate, thus explaining the observed slowing of the spreading. PMID- 10479415 TI - Light Scattering Study of Magnetic Latex Particles and Their Interaction with Polyelectrolytes. AB - Commercially available latex particles with superparamagnetic inclusions have been characterized by polarized and depolarized dynamic light scattering. The particles are spherical, but the larger ones (R(g) approximately 0.1 um) with many magnetic inclusions behave in dynamic light scattering experiments like symmetric tops while the smaller particles do not. The as-received materials had a broad size distribution, but more narrowly distributed particles were prepared by selective filtration for use in probe diffusion and stability studies. The interaction of these negatively charged particles with a linear polyelectrolyte, negatively charged sodium polystyrene sulfonate, was studied by static and dynamic light scattering. Depolarized dynamic light scattering is particularly sensitive to changes in the stability of the colloids or interaction with the polyelectrolyte. The polyelectrolyte can prevent or reverse salt-induced aggregation of the superparamagnetic latex particles. Fluorescence photobleaching recovery experiments to monitor the mobility of fluorescently labeled polyelectrolyte suggest that it is not bound to the latex particles; apparently, a mechanism other than steric stabilization is involved. The dependence of the diffusion of the superparamagnetic latex particles upon added salt is discussed in terms of the ion atmosphere surrounding the polyelectrolyte. PMID- 10479416 TI - Hydrodynamics of Fractal Aggregates with Radially Varying Permeability. AB - Hydrodynamic properties of fractal aggregates with radially varying permeability are investigated in terms of two parameters: radius of equivalent solid sphere experiencing the same drag as the aggregate (Omega*radius of aggregate) and fluid collection efficiency (eta) of the aggregate. Resistance to the fluid flow through the aggregate is predicted to increase with increasing fractal dimension, while the fluid collection efficiency is expected to decrease. It is shown that a reasonable estimate of the fluid drag force on a fractal aggregate may be obtained by assigning a constant volume-averaged porosity to the aggregate and using any of the expressions available in the literature for aggregates with uniform permeability. The two hydrodynamic parameters, Omega and eta, are used to modify the existing expressions for interactions between solid spheres to account for the porous nature of aggregates and thus calculate the collision rate kernels for interacting aggregates. The ratio of hydrodynamic radius of an aggregate to its radius of gyration predicted by the proposed model was in reasonable agreement with an experimental value reported in the literature. PMID- 10479417 TI - Surface Rheological Data for a Polymeric Surfactant Using a Pulsed Drop Rheometer. AB - Measurements of dynamic interfacial tension of adsorbed layers of the oil-soluble polymeric surfactant E5 have been made using a pulsed drop rheometer. The pulsed drop rheometer is based on the instantaneous expansion of a water droplet in oil. After perturbation an interfacial relaxation occurs and is followed from the drop profile. The difference in pressure across a curved interface and the interfacial tension are directly related. The decay of pressure change, and hence the interfacial tension decay, is followed as a function of time using a sensitive pressure transducer. Concentrations of E5 above and below the CMC were investigated at the n-decane/water and Isopar M/water interfaces. The interfacial tension decays obtained were fitted to known relaxation mechanisms. Fourier transforms were calculated over a complete frequency spectrum to obtain the dilational elasticity and viscosity. Above the CMC, the interfacial relaxation of E5 at both the n-decane/water and Isopar M/water interfaces was shown to be due to the diffusion of micelles to the interface and the subsequent lowering of the interfacial tension. From the calculated diffusion coefficient and micelle size, the micellar aggregation number could be calculated. Below the CMC, both diffusion and reorientation contribute to the interfacial relaxation. It was not possible to determine the parameters for each process because the characteristic frequencies for the two processes are of similar magnitude. PMID- 10479418 TI - Effects of Hydrophilization and Immobilization on the Interfacial Behavior of Immunoglobulins. AB - The adsorption and immobilization of rabbit anti-human immunoglobulin (rabbit IgG), as well as the effects of rinsing with buffer and addition of bovine serum albumin (BSA) or human IgG on the amount and reactivity of bound rabbit IgG, were investigated with ellipsometry, total internal reflection fluorescence spectroscopy (TIRF), and enzyme immuno assay (EIA). It was found that although rabbit IgG readily adsorbs at hydrophobic hexamethyldisiloxane (HMDSO) plasma polymer surfaces, a substantial fraction of the adsorbed protein molecules is desorbed upon rinsing with buffer. BSA was found to adsorb readily at the surfaces obtained after rinsing, although also this protein desorbed to a large extent (about 60%) upon further rinsing with buffer. The adsorption of BSA causes a further reduction in the amount of rabbit IgG adsorbed. Immobilization of rabbit IgG to acrylic acid (AA) plasma polymer surfaces, achieved by covalent coupling via a strongly adsorbed PEG-PEI copolymer, was found to overcome the problem of the desorption of rabbit IgG upon rinsing with buffer or addition of BSA. Furthermore, nonspecific adsorption was virtually absent after immobilization. However, covalently bound rabbit IgG reacted strongly with human IgG, as observed by ellipsometry, TIRF, and EIA. The immobilization of rabbit IgG to hydrophilized surfaces was found to facilitate the interpretation of EIA results. PMID- 10479419 TI - NMR Study of Phase Transitions in Pure Water and Binary H(2)O/HNO(3) Films Adsorbed on Surface of Pyrogenic Silica. AB - Pyrogenic silica (aerosil) was employed as host within which the phase transitions in the adsorbed pure water and binary H(2)O/HNO(3) films have been studied with NMR spectroscopy. The median freezing temperature and freezing temperature region were shown to be highly sensitive both to the average thickness of the adsorbed films and to the amount of adsorbed nitric acid. The molar concentration of nitric acid in the adsorbed films was found to be very small, on the order of 10(-3)-10(-2) (M/liter). The concentration was found to be greater in the layers adjacent to the surface of silica and sharply decreases with distance from the surface. The difference between the median freezing temperatures for adsorbed pure water and for the binary system was found to be about 9 K for films of equal thickness. This is about 150 times greater than the difference between the freezing temperatures of bulk pure water and a solution with the same concentration of nitric acid. PMID- 10479420 TI - Quantification of Spatial Correlation in Porous Media and Its Effect on Mercury Porosimetry. AB - In many porous media the grains are packed in a disordered manner, rather than in regular lattices. Theoretical treatments of the properties of these media often assume that because there is no regular lattice, the pore space between grains is completely spatially disordered. Here we present an analysis of a real granular medium (a close packing of equal spheres) which shows that, contrary to the popular assumption, the pore space is spatially correlated. The origin of this pore space correlation is the strong spatial correlation of grain locations, which is a feature of all dense granular media. Our analysis relies on physically representative network models of the pore space constructed from knowledge of the grain locations. Simulated drainage experiments on these networks agree with mercury porosimetry experiments in simple sandstones, whereas simulations in uncorrelated but otherwise identical networks do not. Thus the spatial correlation inherent in the pore space of simple porous media significantly affects mercury porosimetry. Deriving pore size distributions from mercury porosimetry without considering spatial correlation can give misleading results. The likelihood of error is compounded if such pore size distributions are used to estimate transport coefficients such as permeability, diffusivity, and electrical conductivity. PMID- 10479421 TI - Synthesis and Behavior at the Air-Water Interface of Fluorinated Nonionic Surfactants Containing Two Methylated Polyoxyethylene Moieties. AB - The nonionic surfactants of the type polyoxyethylated monomethylated and containing a fluorinated tail were synthesized from 2-(F-alkyl)ethylamines. These compounds have two polyethoxylated tails with a perfectly defined number of oxyethylene motifs. Compared with their bipodal homologs within the F-alkylated series, these new structures were easily obtained from commercial raw materials which are the 2-(F-alkyl)ethyliodides. The evaluation of their behavior at the air-water interface has been studied from measurements of surface tension versus variation of concentration. This clearly shows the variation of the critical micelle concentration (c.m.c.) from 1.0.10(-5) to 2.5.10(-3) mol.liter(-1) and of the surface area per surfactant molecule versus studied structure. The N,N dipolyethoxylene methoxyether of 2-(F-alkyl)ethylamines so realized exhibit noteworthy properties as nonionic fluorinated surfactants. PMID- 10479422 TI - Adsorption of EDTA and Metal-EDTA Complexes onto Goethite. AB - The adsorption characteristics of a variety of divalent and trivalent metal-EDTA complexes onto goethite (alpha-FeOOH) were examined in aqueous solution. Uncomplexed EDTA is adsorbed as a binuclear complex at low pH and as a mononuclear complex at high pH. Adsorption is ligand-like with a high extent of adsorption at low pH. The process can be described by formation of inner-sphere complexes by the surface complexation model with constant capacitance. The EDTA complexes of the divalent metals Ca, Zn, Ni, Cu, Co(II), and Pb, which are quinquedentate in solution (free donor atoms bound to the metal ion), all showed the same ligand-like adsorption behavior. Their adsorption as a function of pH and concentration can be described by the formation of one type of ternary surface complex and can be fitted with the same equilibrium constant. Pd(II)EDTA, which is bidentate or quadridentate in solution, is adsorbed more strongly, but also in a ligand-like manner. The EDTA complexes of the trivalent metals LaEDTA and BiEDTA are adsorbed very strongly over the whole pH range. The sexidentate complex of Co(III) is weakly adsorbed at low pH outer-spherically, i.e., by electrostatic interaction only. Fe(III)EDTA is weakly adsorbed over the whole pH range with a predominant nonspecific surface complex at low pH and a specific complex at high pH. PMID- 10479423 TI - Surface Complexation at the TiO(2) (anatase)/Aqueous Solution Interface: Chemisorption of Catechol. AB - Catechol adsorbs at the TiO(2) (anatase)/aqueous solution interface forming inner sphere surface complexes. The UV-visible differential reflectance spectrum of surface titanium-catecholate complexes presents a band centered at 420 nm which corresponds to the ligand to metal charge transfer transition within the surface complexes. At pH values below pK(a1), the surface excess of catechol is almost insensitive toward pH and presents a Langmuirian dependence with the concentration of uncomplexed catechol. The ratio Gamma(max):N(S) (N(S) being the measured density of available OH surface groups) indicates a prevailing 1 to 2 ligand exchange adsorption stoichiometry. In the range pH >/= pK(a1), the catechol surface excess decreases markedly with increasing pH. Formation of 1 to 1 surface complexes produces an excess of negative surface charge that is revealed by the shift of the iep to lower pH values. The reported data, which are supplemented with information on the charging behavior of TiO(2) suspended in indifferent electrolyte solutions, are interpreted in terms of the multi-site surface complexation model. In this model, two types of surface OH groups are considered: identical withTiOH(1/3-) and identical withOH(1/3+). Although both surface groups undergo protonation-deprotonation reactions, only identical withTiOH(1/3-) are prone to chemisorption. PMID- 10479424 TI - Orientation Distribution and Electrophoretic Motions of Rod-like Particles in a Capillary. AB - We consider motions of charged rod-like particles in a capillary under the action of externally applied electric field as a model problem for the electrophoresis through a capillary. Since the electrophoretic velocity through the capillary is dependent upon the particle orientation relative to the electric field, the probability distribution function for the particle orientation is determined from the Smoluchowski equation. In the statistical equation considered here, the random rotary Brownian potential is balanced with the potentials from both electrical and hydrodynamic origins. First, the dipole moment associated with the asymmetrical distribution of particle surface charges and that induced by the external electric field are calculated by utilizing slender body theory. The results are strictly valid under the conditions that the contribution from double layer distortion is negligible, which is typical of charged macromolecules in aqueous media. The dipole moments are expressed in terms of the particle charge (or zeta potential) distribution, external field strength, and aspect ratio of the particle. The two distinct dipole moments incorporate into the Smoluchowski equation for the particle orientation distribution which in turn determines the electrophoretic mobility. Finally, the particle velocity in a capillary is simply given as superposition of the electrophoretic velocity and the electroosmotic velocity which is driven by the surface charges on the capillary wall. PMID- 10479425 TI - 3H-Indoles in Cetyltrimethylammonium Bromide (CTAB) Micelles and Water: Spectroscopy and Photophysics at Various Temperatures. AB - An absorption and fluorescence spectral study has been carried out for four molecules: 2-[(p-amino)phenyl]-3,3-dimethyl-3H-indole (1), 2-[(p dimethylamino)phenyl]-3,3-dimethyl-3H-indole (2), 2-[(p-amino)phenyl]-3,3 dimethyl-5-carboethoxy-3H-indole (3), and 2-[(p-dimethylamino)phenyl]-3,3 dimethyl-5-carboethoxy-3H-indole (4) in cetyltrimethylammonium bromide (CTAB) and water solutions. Arrhenius plots for the nonradiative decay processes competing with fluorescence are nonlinear in CTAB. This is ascribed to the fact that the probes are locked into a site where the twisting of the phenyl moiety is nearly inhibited in CTAB at room temperature. The binding constants increase in an increasing order of the molecular size (exponential growth) thereby showing that hydrophobic effects are the driving force behind the solubilization of these molecules in CTAB. Calculation of transfer free energies has confirmed the importance of the hydrophobic effect compared to the Coulombic interactions. The neutral-monocation equilibrium is lowered in presence of CTAB. PMID- 10479426 TI - The Drop Volume Method for Interfacial Tension Determination: An Error Analysis. AB - An error analysis of the drop volume method of determination of surface or interfacial tension is presented. It is shown that the presence of the empirical correction term may lead to either a decrease or an increase in the final uncertainty of the calculated tension. Recommendations to maximize the precision of measurement are made. It is further shown that the systematic error due to the correction term is less than 0.04%; under the conditions recommended to minimize the statistical uncertainty, the systematic error should be less than half this figure. Tabulations of recommended values of the correction function are given. PMID- 10479427 TI - Anisotropy Changes of a Fluorescent Probe during the Micellar Growth and Clouding of a Nonionic Detergent. AB - The effects of temperature and Triton X-114 (TX-114) concentration on the fluorescence anisotropy of perylene were investigated before and after detergent clouding. The measured anisotropy values were used to estimate the microviscosity of the micellar interior. In the lower detergent concentration range, an anisotropy maximum was observed at the critical micelle concentration (CMC), while the values decreased in the range immediately above the CMC. This was ascribed to the micellar volume increase, which, in the case of TX-114, was not accompanied by a more ordered internal environment. A gradual decrease of anisotropy and microviscosity with increasing temperature below the cloud point was observed. At the cloud point, no abrupt changes were found to occur. Compared to detergents with more flexible hydrophobic moieties, TX-114 micelles have a relatively ordered micellar interior indicated by the microviscosity and calculated fusion energy values. In the separated micellar phase formed after clouding, the probe anisotropy increased as water was eliminated at higher temperatures. PMID- 10479428 TI - Surface Functionalization of Polyethylene Terephthalate Film and Membranes by Controlled Wet Chemistry. AB - Surface functionalization of polyethylene terephthalate film and track-etched microporous membranes was performed using organic synthesis at the solid-liquid interface. The hydroxyl chain ends, naturally occurring in the membranes or created by basic hydrolysis in the film, were activated by reaction with p toluenesulfonyl chloride and then coupled to [4,5-(3)H]lysine used as a model of water-soluble bioactive molecules. The ratios of derivatization were determined by liquid scintillation counting of the radioactivity associated with the samples. The hydroxyl chain ends were further reacted with 4,4'-methylene di(phenyl isocyanate); the resulting surfaces equipped with sensitive isocyanate spacers were assayed as before using [4,5-(3)H]lysine. The ranges of derivatization were from 5 to 60 pmol of fixed amino acid per open surface unit (cm(2)). Some samples were also analyzed by X-ray photoelectron spectroscopy. PMID- 10479429 TI - Adsorption of Single Anionic Surfactants on Hydrophobic Surfaces. AB - The adsorption of a series of linear alkyl sulfonates and linear alkyl sulfates on sulfonated polystyrene latexes of various surface charge densities was studied to provide a better understanding of the effects of surface charge on the adsorption of anionic surfactants. Direct measurement of the adsorbed amount via ion chromatography provided greater accuracy and precision than do indirect methods such as soap titration. Results indicate that in the region of surface charge densities between -3 and -7 uC/cm(2) the effects of surface potential on adsorbed amount are more complex than has been described in the past. Previous work hypothesized a steady decrease in adsorbed amount with increasing surface charge due to increased electrostatic repulsions. The data obtained in this investigation indicate local increases in the adsorbed amount, possibly a result of configurational rearrangements of the adsorbed surfactant leading to an increase in surface area available for adsorption. PMID- 10479430 TI - Scintigraphic Study of Local Flux and Osmotic Pressure Distributions in Ultrafiltration of Blood and Plasma. AB - We have investigated the spatial variation of local protein concentration and filtration flux by a scintigraphic technique in the ultrafiltration of bovine albumin solutions and blood. The feed was mixed with (99m)Tc albumin macroaggregates and circulated through a polysulfone 30,000 MWCO hollow fiber filter placed in the field of a gamma-camera. Concentration profiles c(b) (x) were reconstructed from scintigraphic images and the local ultrafiltration flux was calculated by differentiating c(b) (x) and using mass conservation. Tests were run at various inlet shear rates from 472 to 1415 s(-1) and under two different filtration regimes: no net filtration (permeate valve closed) and large filtration (below the pressure independent plateau). The data confirm the filtration decay from the filter inlet to outlet but an unexpected result is the presence of high retrofiltration in the downstream part of the filter length in the case of large filtration. This retrofiltration can be explained by a high osmotic pressure at the membrane created by the protein polarization concentration. Assuming a constant pressure gradient along the fibers, it is possible to estimate the local osmotic pressure at the onset of retrofiltration and to infer from it the protein concentration at the membrane, which is found to vary from 170 to 250 g/liter when gamma(w) increases. Similar experiments were run with blood and a microfiltration membrane (0.55-um pores). In that case no retrofiltration was obtained, which confirms our explanation since in this case the polarization layer is composed of red cells which exert no osmotic pressure. PMID- 10479431 TI - Monte Carlo Simulation of Controlled Colloid Growth by Homo- and Heterocoagulation in Two Dimensions. AB - The kinetics of two dimensional homo- and heterocoagulation of particles of equal size were investigated by Monte Carlo simulation using a square lattice and particles with a functionality of four. In both processes, a size-dependent cluster aggregation ability gamma allowed simulation of diffusional and reactional limited processes. In heterocoagulation, determination of the effects of the relative concentration x of the particles A and B and the cluster aggregation ability on the aggregation rate provided information concerning the influence of the frequency of unsuccessful encounters between identical particles on the temporal increases in the weight S(t, x, gamma) and number N(t, x, gamma) average sizes of the clusters. Dynamic scaling exponents for the temporal variations of the average cluster sizes were found to be linear or power law functions of the composition x. PMID- 10479432 TI - Internal Structure Analysis of Monodispersed Pseudocubic Hematite Particles by Electron Microscopy. AB - Morphology and internal structure of monodispersed pseudocubic hematite (alpha Fe(2)O(3)) particles produced by the gel-sol method were investigated in detail through high-resolution electron microscopy on their thin sections prepared with an ultramicrotome. It was confirmed that the c-axis of a particle corresponded to the longest diagonal axis of the particle. High-resolution electron micrographs of thin sections directly revealed the polycrystallinity of the particles and clearly showed the arrangement of the subcrystals and their crystallographic orientations. The subcrystals near the surface of a particle were of a rectangular shape bound by the $ ?{01?overline{1}2?} $ planes, and their width was about 12-16 nm with small variation of the length depending on the position in a particle. The subcrystals were radially developed from the center of a particle in all directions, but most preferentially in the longest diagonal axis of a particle. Besides, it was found from EDX analysis that the adsorbed Cl(-) ions as a shape controller were incorporated into the pseudocubic hematite particles during their growth. PMID- 10479433 TI - A Stray Field Magnetic Resonance Imaging Study of the Drying of Sodium Silicate Films. AB - Stray field magnetic resonance imaging (STRAFI) is shown to be highly suited to the study of drying processes in thin films. Sodium silicate films have been chosen as a model system exhibiting many of the properties of film drying in general. Films have been dried, as a function of temperature in the range 22 to 62 degrees C, down to water contents of order 28% by weight, at which stage the film is glassy. The experimental results have been quantitatively analyzed by treating the drying film as a colloidal solution. The results suggest that the localized hydrogen spin-spin relaxation time, and hence the mobility of the water in the films is independent of the drying regime and depends primarily on the local water concentration. PMID- 10479434 TI - Comparison of Adsorption and Conformation of Hemoglobin and Myoglobin on Various Inorganic Ultrafine Particles. AB - Human hemoglobin and horse myoglobin were adsorbed on the two types of ultrafine silica (silica 1 and 2), titania, and zirconia particles under various conditions. The order of the affinity of the ultrafine particles to these proteins was silica 1 < silica 2 < zirconia = titania under the same adsorption condition. The conformational changes in these proteins upon adsorption and desorption were monitored by both circular dichroism and absorbance spectra. In both spectrum measurements, a similar tendency was observed, indicating that the adsorption of proteins causes the changes not only in the secondary structure but also in the tertiary structure. In all the particles, the extent of conformational changes upon adsorption increased with decreasing pH and adsorption amount. On the other hand, the extent of conformational changes was significantly affected by the particle type (its order was silica 1 < silica 2 < zirconia = titania) under the same adsorption condition. The percentage of desorption and the extent of refolding of desorbed proteins were lower when the extent of conformational changes in the adsorbed states was higher. These results clearly demonstrate that higher affinity between particle surfaces and proteins causes larger conformational changes and consequently leads to higher irreversibility in adsorption and conformational states. PMID- 10479435 TI - Adsorption Behavior of Cationic and Nonionic Surfactant Mixtures at the Alumina Water Interface. AB - The solution and interfacial behavior of surfactant mixtures was investigated using a cationic surfactant, tetradecyl trimethyl ammonium chloride (TTAC), and a nonionic surfactant, pentadecylethoxylated nonyl phenol (NP-15), with alumina as the substrate. The cationic TTAC adsorbed at the alumina-water interface as a result of electrostatic attraction at pH approximately 10 whereas the nonionic NP 15 did not. Interestingly, in the mixed surfactant system, the tetradecyl trimethyl ammonium chloride forced adsorption of the NP-15 as a result of hydrophobic interactions between the adsorbed surfactant chains at the alumina water interface. The adsorption behavior was dependent upon the ratio of the two surfactants in the mixture as well as the order of addition. With an increase in TTAC content, the adsorption density of NP-15 increased and the isotherm shifted to lower surfactant concentrations. The adsorption of TTAC decreased under conditions of saturation adsorption due to the bulkiness of the coadsorbed NP-15 as well as competition for common adsorption sites. However, below saturation adsorption conditions, the adsorption of TTAC was increased due to synergistic interactions between the cationic and nonionic heads leading to reduced repulsion among the cationic headgroups. Zeta potential measurements showed that, with an increase of NP-15 in the mixtures, the positive charge of the TTAC was partially screened by the coadsorbed NP-15. Surface tension of the surfactant mixtures was also measured, and regular solution theory was used to model the interactions between the surfactants. Monomer concentrations obtained using the theory were correlated with the adsorption tendencies and it was seen that TTAC adsorption corresponded with its monomer concentration but that of NP-15 did not, suggesting the need for adequate theories for interactions between surfactants. PMID- 10479436 TI - Effect of Additives on the $ ?hbox{I}_{2} + ?hbox{I}^{-} ?leftrightarrow ?hbox{I}^{-}_{3} $ Reaction in Micellar Solutions of Sodium Dodecylsulfate: Partitioning and Solubilization Site of Iodine. AB - The partition constants (K(p)) of iodine between sodium dodecylsulfate micelles and the external media have been measured in the presence of water-soluble (sodium sulfate) and micelle-soluble (n-hexane and n-hexanol) additives. A spectrophotometric method based in the analysis of the alterations provoked by the presence of the surfactant plus the additives upon the reversible reaction taking place between iodine and iodide to give tri-iodide ions was employed (Abuin, E., and Lissi, E., J. Chem. Ed. 69, 340 (1992)). The effect of the additives upon the n-octanol/water partitioning (K(o/w)) of iodine was also determined. The results obtained for K(p) and its relationship with the K(o/w) values are indicative of a preferential localization of iodine at the surface of the micelles. PMID- 10479437 TI - Microscopic and Macroscopic Dynamic Interface Shapes and the Interpretation of Dynamic Contact Angles. AB - We have studied shapes of dynamic fluid interfaces at distances 0.01. Our measured dynamic contact angle parameter extracted by fitting the analysis with viscous deformation to the shape near the moving contact line coincides with the contact angle of the static-like shape far from the contact line. We measure and explain the discrepancy between this dynamic contact angle parameter and the apparent contact angles based on meniscus or apex heights. Our observations of viscous effects at large distances from the contact line have implications for dynamic contact angle measurements in capillary tubes. PMID- 10479438 TI - Selective Isolation of DNA or RNA Using Single-Stranded DNA Affinity Latex Particles. AB - We developed latex particles with a diameter of about 0.22 um on which single stranded (ss) DNA was covalently coupled to select or enrich its complementary DNA or mRNA. DNA was first covalently coupled to the latex particles in the double-stranded (ds) form with both blunt and protruding ends. More than 80% of the dsDNA was coupled through the ssDNA stretch at its protruding end. The presence of NaCl in the immobilization reaction severely inhibited DNA from coupling to the particles. The particles were then treated with alkali or heated to denature the dsDNA and sedimented by a brief centrifugation to yield ssDNA immobilized particles. They allowed the selective and efficient isolation of a desired RNA from total cellular RNA. PMID- 10479439 TI - Optical Properties of Nonaqueous Suspensions in Electric Fields. AB - The optical properties of suspensions consisting of pigmented shell/polymer core composite particles dispersed in a silicone oil were studied in electric fields. In the absence of electric fields, the transmittance shows an exponential decay with sample thickness and particle concentration. The particles may be randomly dispersed in the medium. The application of electric field leads to an increase in the transmittance of suspensions. The clearing effect is very striking, especially for concentrated suspensions in a thick cell. In electric fields, the transmittance is almost independent of thickness in the range 0.15-1.1 mm and linearly decreases with increasing particle concentration. Microscopic observation shows that many discrete clusters are formed in electric fields. Based on the analysis of dipole-dipole interactions, the clusters may be composed of fully developed chains spanning the electrodes. Since the columns are aligned in the direction parallel to the optical path, the transmittance is not affected by the cell thickness. In addition, because of the increase in the cross section of the column, the transmittance shows a linear dependence on the particle concentration. The optical properties of suspensions can be explained by the column formation of particles in electric fields. PMID- 10479440 TI - The Dual Role of Fibrinogen as Inhibitor and Nucleator of Calcium Phosphate Phases: The Importance of Structure. AB - Constant composition and free drift methods have been used to investigate the abilities of human serum albumin (HSA) and fibrinogen to influence calcium phosphate precipitation. Both molecules inhibit hydroxyapatite (HAP) crystal growth when present in the solution phase. Fibrinogen, when immobilized at hydrophobicized germanium or silica surfaces, is able to nucleate calcium phosphate phases; at clean germanium or silica surfaces, it appears to be inactive. The apparent configuration of fibrinogen molecules at germanium or silica surfaces on which octadecyltrichlorosilane (OTS) was deposited probably exposes more negative sites for participation in nucleation. PMID- 10479441 TI - Structure and Bonding of Orthophosphate Ions at the Iron Oxide-Aqueous Interface. AB - The surface speciation of orthophosphate ions on goethite has been studied as a function of pH, time, total phosphate concentration, and ionic medium by means of diffuse reflectance FTIR spectroscopy. The samples were prepared in accordance with a distribution diagram of surface species as calculated from thermodynamic data. In agreement with the thermodynamic model three dominating surface complexes could be distinguished with IR spectroscopy, and the relative distribution of the species was shown to be primarily a function of pH. The IR characteristics of the individual surface complexes were indicative of molecular symmetries of the PO(4) unit of C(3v), C(2v), and C(3v), respectively. This was concluded to be incompatible with the bidentate, bridging structural model previously suggested. Instead, the IR data are in good agreement with a monodentate coordination of phosphate to the surfaces, where the three surface complexes only differ in the degree of protonation. A comparison between the adsorption behavior of phosphate on goethite and hematite was also made. Here the importance of the aqueous stability of the adsorbent on the adsorption mechanism was shown. PMID- 10479442 TI - Electrostatic Protein-Protein Interactions: Comparison of Point-Dipole and Finite Length Dipole Potentials of Mean Force. AB - Based on summation of Coulombic interactions, a model is developed for finite length dipole potentials of mean force in a salt-free dielectric continuum. Point dipole and finite-length dipole potentials of mean force are compared for protein protein interactions using parameters for bovine alpha-chymotrypsin. The two approximations made in the commonly used analytical point-dipole potentials of mean force are not valid at distances near contact. Relative to the finite-length dipole model, the high-temperature approximation overpredicts, and the point dipole approximation underpredicts charge-dipole and dipole-dipole attractions. PMID- 10479443 TI - The Dielectric Response of a Dilute Suspension of Semiconducting Particles. AB - The aim of this note is to draw attention to the interesting electrical properties of aqueous suspensions of semiconducting particles. The charge carriers in these particles have a very high mobility, and, as a result, large polarization currents are set up when an external field is applied. For spherical particles this has the effect of causing the particles to behave like dielectrics with an enhanced permittivity epsilon*. The value of the effective permittivity depends on the ratio of the particle radius to the internal Debye length of the semiconductor. A formula for epsilon* is obtained in the case when the equilibrium charge carrier distribution is uniform over the particle. PMID- 10479444 TI - Gastrulation in zebrafish: what mutants teach us. AB - A major approach to the study of development is to compare the phenotypes of normal and mutant individuals for a given genetic locus. Understanding the development of a complex metazoan therefore requires examination of many mutants. Relatively few organisms are being studied this way, and zebrafish is currently the best example of a vertebrate for which large-scale mutagenesis screens have successfully been carried out. The number of genes mutated in zebrafish that have been cloned expands rapidly, bringing new insights into a number of developmental pathways operating in vertebrates. Here, we discuss work on zebrafish mutants affecting gastrulation and patterning of the early embryo. Gastrulation is orchestrated by the dorsal organizer, which forms in a region where maternally derived beta-catenin signaling is active. Mutation in the zygotic homeobox gene bozozok disrupts the organizer genetic program and leads to severe axial deficiencies, indicating that this gene is a functional target of beta-catenin signaling. Once established, the organizer releases inhibitors of ventralizing signals, such as BMPs, and promotes dorsoanterior fates within all germ layers. In zebrafish, several mutations affecting dorsal-ventral (D/V) patterning inactivate genes functioning in the BMP pathway, stressing the central role of this pathway in the gastrula embryo. Cells derived from the organizer differentiate into several axial structures, such as notochord and prechordal mesoderm, which are thought to induce various fates in adjacent tissues, such as the floor plate, after the completion of gastrulation. Studies with mutants in nodal-related genes, in one-eyed pinhead, which is required for nodal signaling, and in the Notch pathway reveal that midline cell fate specification is, in fact, initiated during gastrulation. Furthermore, the organizer coordinates morphogenetic movements, and zebrafish mutants in T-box mesoderm-specific genes help clarify the mechanism of convergence movements required for the formation of axial and paraxial mesoderm. PMID- 10479445 TI - Membrane hyperpolarization by sperm-activating and -attracting factor increases cAMP level and activates sperm motility in the ascidian Ciona intestinalis. AB - In the ascidian Ciona intestinalis (and C. savignyi), sperm-activating and attracting factor (SAAF) is released from the egg at fertilization and stimulates both Ca(2+) influx and a transient increase in cAMP level of the sperm, leading to the activation of sperm motility (M. Yoshida et al., 1994, Dev. Growth Differ. 36, 589-595). In this paper we show in C. intestinalis that valinomycin, a potassium-selective ionophore, as well as SAAF, activated sperm motility, and this activation was suppressed by extracellular high K(+). Membrane potential measurements showed that both SAAF and valinomycin increase K(+) permeability of sperm and induce membrane hyperpolarization, the amplitude of which depends on the external K(+) concentration. The membrane potential and intracellular K(+) concentration of Ciona sperm without SAAF were estimated to be about -50 mV and 560 +/- 40 mM, respectively. After treatment with SAAF or valinomycin the membrane potential became almost equal to the equilibrium potential of K(+) (-100 mV), and the cAMP level increased in artificial seawater. A potent voltage dependent K(+) channel blocker, MCD peptide, at the concentration of 10 microM blocked SAAF-induced hyperpolarization of the cells, increase in cAMP, and sperm motility. These results suggest that membrane hyperpolarization produced by the opening of K(+) channels elevates cAMP synthesis and leads to the activation of sperm motility in Ciona. PMID- 10479446 TI - Ascidian tail formation requires caudal function. AB - Although the tail is one of the major characteristics of animals of the phylum Chordata, evolutionary aspects of the molecular mechanisms involved in its formation are not clear. To obtain insights into these issues, we have isolated and investigated the caudal gene of an ascidian, one of the lower animal groups among chordates. Ascidian caudal is expressed from the midgastrula stage onward in the lateral walls of the posterior neural tube cell lineage and also in the posterior epidermal cells from the neurula stage. Thus, ascidian caudal expression is restricted to the ectoderm of a tail-forming region throughout embryogenesis. Suppression of caudal function by an antisense oligonucleotide or a dominant negative construct caused inhibition of the cell movement required for tail formation. Overexpression of wild-type caudal mRNA in an ascidian animal cap, an animal half explant prepared at the eight-cell stage, caused elongation of the cap. Furthermore, Xenopus embryos injected with dominant negative ascidian caudal exhibited defects in elongation, suggesting a conserved caudal function among chordates. These results indicate that caudal function is required for chordate tail formation and may play a key role in its evolution. PMID- 10479447 TI - Calcium signaling in the developing Xenopus myotome. AB - Embryonic Xenopus myocytes generate spontaneous calcium (Ca(2+)) transients during differentiation in culture. Suppression of these transients disrupts myofibril organization and the formation of sarcomeres through an identified signal transduction cascade. Since transients often occur during myocyte polarization and migration in culture, we hypothesized they might play additional roles in vivo during tissue formation. We have tested this hypothesis by examining Ca(2+) dynamics in the intact Xenopus paraxial mesoderm as it differentiates into the mature myotome. We find that Ca(2+) transients occur in cells of the developing myotome with characteristics remarkably similar to those in cultured myocytes. Transients produced within the myotome are correlated with somitogenesis as well as myocyte maturation. Since transients arise from intracellular stores in cultured myocytes, we examined the functional distribution of both IP(3) and ryanodine receptors in the intact myotome by eliciting Ca(2+) elevations in response to photorelease of caged IP(3) and superfusion of caffeine, respectively. As in culture, transients in vivo depend on Ca(2+) release from ryanodine receptor (RyR) stores, and blocking RyR during development interferes with somite maturation. PMID- 10479448 TI - Mammalian BMP-1/Tolloid-related metalloproteinases, including novel family member mammalian Tolloid-like 2, have differential enzymatic activities and distributions of expression relevant to patterning and skeletogenesis. AB - Vertebrate bone morphogenetic protein 1 (BMP-1) and Drosophila Tolloid (TLD) are prototypes of a family of metalloproteases with important roles in various developmental events. BMP-1 affects morphogenesis, at least partly, via biosynthetic processing of fibrillar collagens, while TLD affects dorsal-ventral patterning by releasing TGFbeta-like ligands from latent complexes with the secreted protein Short Gastrulation (SOG). Here, in a screen for additional mammalian members of this family of developmental proteases, we identify novel family member mammalian Tolloid-like 2 (mTLL-2) and compare enzymatic activities and expression domains of all four known mammalian BMP-1/TLD-like proteases [BMP 1, mammalian Tolloid (mTLD), mammalian Tolloid-like 1 (mTLL-1), and mTLL-2]. Despite high sequence similarities, distinct differences are shown in ability to process fibrillar collagen precursors and to cleave Chordin, the vertebrate orthologue of SOG. As previously demonstrated for BMP-1 and mTLD, mTLL-1 is shown to specifically process procollagen C-propeptides at the physiologically relevant site, while mTLL-2 is shown to lack this activity. BMP-1 and mTLL-1 are shown to cleave Chordin, at sites similar to procollagen C-propeptide cleavage sites, and to counteract dorsalizing effects of Chordin upon overexpression in Xenopus embryos. Proteases mTLD and mTLL-2 do not cleave Chordin. Differences in enzymatic activities and expression domains of the four proteases suggest BMP-1 as the major Chordin antagonist in early mammalian embryogenesis and in pre- and postnatal skeletogenesis. PMID- 10479449 TI - LOLA has the properties of a master regulator of axon-target interaction for SNb motor axons of Drosophila. AB - The proper pathfinding and target recognition of an axon requires the precisely choreographed expression of a multitude of guidance factors: instructive and permissive, positive and negative, and secreted and membrane bound. We show here that the transcription factor LOLA is required for pathfinding and targeting of the SNb motor nerve in Drosophila. We also show that lola is a dose-dependent regulator of SNb development: by varying the expression of one lola isoform we can progressively titrate the extent of interaction of SNb motor axons with their target muscles, from no interaction at all, through wild-type patterning, to apparent hyperinnervation. The phenotypes we observe from altered expression of LOLA suggest that this protein may help orchestrate the coordinated expression of the genes required for faithful SNb development. PMID- 10479450 TI - Multiple roles for activin-like kinase-2 signaling during mouse embryogenesis. AB - The members of the transforming growth factor-beta (TGF-beta) superfamily are secreted proteins that interact with cell-surface receptors to elicit signals that regulate a variety of biological processes during vertebrate embryogenesis. Alk2, also known as ActRIA, Tsk7L, and SKR1, encodes a type I TGF-beta family receptor for activins and BMP-7. Initially, Alk2 transcripts are detected in the visceral endoderm of gastrula stage mouse embryos, suggesting a signaling role in extraembryonic tissues during development. To study the role of Alk2 during mammalian development, Alk2 mutant mice were generated. After embryonic day 9.5 (E9.5), no homozygous mutants were recovered from heterozygote matings. Homozygous mutants with morphological defects were first detected at E7.0 and were smaller than controls. Morphological and molecular examination demonstrated that Alk2 mutant embryos formed a primitive streak, although abnormally thickened, and were arrested in their development around the late streak stage. These gastrulation defects were rescued in chimeric embryos generated by injection of Alk2 mutant embryonic stem (ES) cells into wild-type blastocysts. This rescue of gastrulation defects was also observed in chimeric embryos generated by aggregation of Alk2 homozygous mutant ES cells with tetraploid wild type embryos. However, at E9.5, these embryos that were completely ES-derived also had defects. In contrast, chimeric embryos generated by injection of wild type ES cells into Alk2 mutant blastocysts did not show rescue of the gastrulation defects. These results suggest that signaling through this type I receptor is essential in extraembryonic tissues at the time of gastrulation for normal mesoderm formation and also suggest that subsequent Alk2 signaling is essential for normal development after gastrulation. PMID- 10479451 TI - An analysis of the early events when oligodendrocyte precursor cells are triggered to differentiate by thyroid hormone, retinoic acid, or PDGF withdrawal. AB - Oligodendrocyte precursor cells withdraw from the cell cycle and terminally differentiate after a limited number of cell divisions. The timing of cell-cycle withdrawal and differentiation is controlled by an intrinsic timer, which consists of a timing component that measures elapsed time and an effector component that arrests the cell cycle and initiates differentiation. The effector component can be triggered by either thyroid hormone (TH) or retinoic acid (RA). In this study we investigate how TH and RA act to trigger differentiation. We show the following: (1) Synthetic retinoids that can inhibit AP-1 transcription factors but do not activate gene transcription cannot trigger the effector mechanism, suggesting that TH and RA do not act only by inhibiting AP-1 activity as previously suggested. (2) Both TH and RA induce a transcriptionally dependent antigenic change in purified precursor cells within 2-4 h. (3) Unexpectedly, even before they differentiate, the precursor cells express ceramide galactosyltransferase (CGT), the enzyme that catalyzes the final step in the synthesis of galactocerebroside, an early marker of oligodendrocyte differentiation. (4) Neither TH nor RA directly activates the transcription of the CGT gene, a number of immediate early genes, or the genes that encode any of the known cyclin-dependent kinase inhibitors. (5) The withdrawal of the mitogen platelet-derived growth factor (PDGF), but not TH or RA treatment, causes a rapid decrease in c-fos, NGFI-A/Krox-24, and cyclin D2 mRNA, even though all three treatments trigger cell-cycle arrest and differentiation. (6) PDGF withdrawal and TH treatment, but not RA treatment, induce an increase in cyclin D3 mRNA within 4 h. Thus, we have not found any early changes in gene expression that occur with all three treatments that trigger oligodendrocyte differentiation. PMID- 10479452 TI - Tissue interactions in the induction of anterior pituitary: role of the ventral diencephalon, mesenchyme, and notochord. AB - Rathke's pouch, the epithelial primordium of the anterior pituitary, differentiates in close topographical and functional association with the ventral diencephalon. It is still not known whether the ventral diencephalon acts as the initial inducer of pituitary development. The roles of the adjacent mesenchyme and notochord, two other tissues located in close proximity to Rathke's pouch, in this process are even less clear. In this report we describe an in vitro experimental system that reproduces the earliest steps of anterior pituitary development. We provide evidence that the ventral diencephalon from 2- to 4-day old chick embryos is able to function as an inducer of pituitary development and can convert early chick embryonic head ectoderm, which is not involved normally in pituitary development, into typical anterior pituitary tissue. This induction is contact-dependent. In our experimental system, there is a requirement for the supporting action of mesenchyme, which is independent of the mesenchyme source. Transplantation of the notochord into the lateral head region of a six-somite chick embryo induces an epithelial invagination, suggesting that the notochord induces the outpouching of the roof of the stomodeal ectoderm that results in formation of Rathke's pouch and causes the close contact between this ectoderm and the ventral diencephalon. Finally, we demonstrate that the ventral diencephalon from e9.5-e11.5 mouse embryos is also an efficient inducer of anterior pituitary differentiation in chick embryonic lateral head ectoderm, suggesting that the mechanism of anterior pituitary induction is conserved between mammals and birds, using the same, or similar, signaling pathways. PMID- 10479453 TI - Loss of ectodermal competence for lateral line placode formation in the direct developing frog Eleutherodactylus coqui. AB - In the direct-developing frog Eleutherodactylus coqui neuromasts and ganglia of the lateral line system never develop. We show here that this absence of the lateral line system, which is evolutionarily derived in anurans, is due to very early changes in development. Ectodermal thickenings, which are typical of lateral line placodes, and from which neuromasts and ganglion cells of the lateral line originate, never form in E. coqui, although other neurogenic placodes are present. Moreover, although NeuroD is expressed in the lateral line placodes of Xenopus laevis, corresponding expression sites are lacking in E. coqui. Heterospecific transplantation experiments show that axolotl ectoderm can be induced to form lateral line placodes after transplantation to E. coqui hosts but that E. coqui ectoderm does not form lateral line placodes on axolotl hosts. This suggests that the loss of the lateral line system in E. coqui is due to the specific loss of ectodermal competence to form lateral line placodes in response to inductive signals. Our results (1) indicate that the competence for lateral line placode formation is distinct and dissociable from the competence to form other neurogenic placodes and (2) support the idea that the lateral line system acts as a module in development and evolution. PMID- 10479454 TI - The frequency of calcium oscillations in mouse eggs at fertilization is modulated by the number of fused sperm. AB - In a variety of calcium signaling systems, the frequency of intracellular calcium oscillations is physiologically important. Probably multiple factors control the frequency of calcium oscillations in the egg after fertilization and many of these remain to be identified. In this study, we present the first rigorous set of data showing that monospermic fertilization is important for setting the physiological calcium oscillation frequency. Recordings in 152 zona-free eggs show that the general pattern of the calcium oscillations is identical in monospermic and polyspermic eggs; however, the oscillation frequency is higher in polyspermic eggs (P < 10(-6)). The frequency of the late oscillations increases with the number of sperm heads incorporated: 5.2 +/- 0.3 spikes per hour (mean +/ SEM; n = 55) in monospermic eggs, 6.6 +/- 0.3 (n = 62) in dispermic eggs, 8.7 +/ 0.7 (n = 23) in trispermic eggs, and 8.9 +/- 0.9 (n = 12) in eggs with four or more sperm heads. The frequency of the early oscillations is also increased in polyspermic eggs. Seventy-eight additional eggs were divided into two groups and inseminated with two different sperm concentrations ("low" and "high") to obtain one group mainly monospermic and the other mainly polyspermic. The two groups of eggs oscillated at different frequencies (P < 10(-5)). These data rule out the possibility of an egg effect in which some eggs would have the dual properties of oscillating faster and of being able to fuse with several sperm cells. These data instead suggest that the sperm modulates the frequency of the oscillations in a dose-dependent manner. PMID- 10479455 TI - NT4/5 mutant mice have deficiency in gustatory papillae and taste bud formation. AB - Neurotrophins are key determinants for controlling the survival of peripheral neurons during development. Brain-derived neurotrophic factor (BDNF) and neurotrophin-4/5 (NT4/5) exert their action through a common trkB receptor but independently support gustatory sensory neurons. To assess the role of NT4/5 during development, we examined the postnatal development and maintenance of fungiform taste buds in mice carrying a deletion of NT4/5. The absence of NT4/5 results in embryonic deficits in gustatory innervation and a reduced number of fungiform papillae at birth. No degenerative deficits of fungiform papillae were observed for the first 3 weeks of postnatal development. However, these remaining fungiform papillae were smaller in appearance and many did not contain taste pores. By postnatal day 60, there was 63% decrease in the number of fungiform papillae, and remaining papillae were smaller in size or modified into filiform like spines. These papillae had either no taste bud or a taste bud with a reduced number of taste cells compared to controls. These findings demonstrate that the NT4/5 gene functions in the maintenance of fungiform gustatory papillae and raises the possibility for an earlier role in development. PMID- 10479456 TI - Retinoic acid hydroxylase (CYP26) is a key enzyme in neuronal differentiation of embryonal carcinoma cells. AB - Besides nuclear retinoid receptors and cellular retinoid binding proteins also retinoic acid (RA)-synthesizing enzymes (using all-trans-retinal as substrate) and RA-catabolizing enzymes (producing hydroxylated products) may explain the specific effects of retinoids. In the past we have established an active role for 4-hydroxy-RA and 4-oxo-RA, which originally were considered to be inactive retinoids, but in fact are highly active modulators of positional specification in Xenopus development. Here we present evidence for a specific role of hydroxylated RA metabolites in the onset of neuronal differentiation. 4-Hydroxy- and 18-hydroxy-RA are products of the hydroxylation of RA by a novel cytochrome P450 (CYP)-type of enzyme, CYP26, expression of which is rapidly induced by RA. P19 embryonal carcinoma (EC) cell lines stably expressing hCYP26 undergo extensive and rapid neuronal differentiation in monolayer at already low concentrations of RA, while normally P19 cells under these conditions differentiate only in endoderm-like cells. Our results indicate that the effects on growth inhibition and RARbeta transactivation of P19 EC cells are mediated directly by RA, while the onset of neuronal differentiation and the subsequent expression of neuronal markers is mediated by hCYP26 via the conversion of RA to its hydroxylated products. PMID- 10479457 TI - HNF-3/forkhead homologue-4 influences lung morphogenesis and respiratory epithelial cell differentiation in vivo. AB - HNF-3/forkhead homologue 4 (HFH-4), a transcription factor of the winged helix/forkhead family, is expressed in various tissues including lung, brain, oviduct, testis, and embryonic kidney. In order to test whether the temporospatial expression of HFH-4 influences lung morphogenesis, HFH-4 was expressed in lungs of transgenic mice under control of the surfactant protein C (SP-C) promoter. The morphology of the lungs from SP-C/HFH-4 embryos (day 18 postconception) was distinctly abnormal, and the severity of the alterations correlated with the level of transgene expression as detected by in situ hybridization. At high levels of expression, HFH-4 altered epithelial cell differentiation and inhibited branching morphogenesis. Atypical cuboidal or columnar cells lined the lung periphery of SP-C/HFH-4 transgenic mice. The atypical epithelial cells seen in the SP-C/HFH-4 mice expressed thyroid transcription factor-1 and hepatocyte nuclear factor 3beta (HNF-3beta). However, surfactant proteins SP-B, SP-C, and Clara cell secretory protein, normally produced by nonciliated epithelial cells in lung parenchyma were lacking. beta Tubulin IV, a marker of ciliated cells, stained the atypical columnar cells produced by expression of high levels of the SP-C/HFH-4 transgene. Ectopic expression of HFH-4 in developing mouse lung altered epithelial cell differentiation and morphology, restricting the expression of markers typical of nonciliated cells of the distal lung parenchyma. PMID- 10479458 TI - Roles for the winged helix transcription factors MF1 and MFH1 in cardiovascular development revealed by nonallelic noncomplementation of null alleles. AB - The murine Mf1 and Mfh1 genes have overlapping patterns of expression in the embryo and encode forkhead/winged helix transcription factors with virtually identical DNA binding domains. Previous studies have shown that Mfh1 null mutants have severe cardiovascular defects, including interruptions and coarctations of the aortic arch and ventricular septal defects (Iida et al., Development 124, 4627-4638, 1997). Here, we show that Mf1(lacZ) homozygous null mutants also have a similar spectrum of cardiovascular abnormalities. Moreover, most embryos doubly heterozygous for Mfh1(tm1) and Mf1(lacZ) die before birth with interruptions and coarctations of the aortic arch, dysgenesis of the aortic and pulmonary valves, ventricular septal defects, and other cardiac anomalies. This nonallelic noncomplementation and the similar patterns of expression of the two genes in the mesenchyme and endothelial cells of the branchial arches, outflow tract, and heart suggest that Mf1 and Mfh1 play interactive roles in the morphogenesis of the cardiovascular system. Implications for the development of human congenital heart defects are discussed. PMID- 10479459 TI - The Drosophila Jak kinase hopscotch is required for multiple developmental processes in the eye. AB - Jak kinases are critical signaling components in hematopoiesis. While a large number of studies have been conducted on the roles of Jak kinases in the hematopoietic cells, much less is known about the requirements for these tyrosine kinases in other tissues. We have used loss of function mutations in the Drosophila Jak kinase Hopscotch (Hop) to determine the role of Hop in eye development. We find that Hop is required for cell proliferation/survival in the eye imaginal disc, for the differentiation of photoreceptor cells, and for the establishment of the equator and of ommatidial polarity. These results indicate that hop activity is required for multiple developmental processes in the eye, both cell-autonomously and nonautonomously. PMID- 10479460 TI - Embryonic expression and function of the chemokine SDF-1 and its receptor, CXCR4. AB - Directed cell movement is integral to both embryogenesis and hematopoiesis. In the adult, the chemokine family of secreted proteins signals migration of hematopoietic cells through G-coupled chemokine receptors. We detected embryonic expression of chemokine receptor messages by RT-PCR with degenerate primers at embryonic day 7.5 (E7.5) or by RNase protection analyses of E8.5 and E12.5 tissues. In all samples, the message encoding CXCR4 was the predominate chemokine receptor detected, particularly at earlier times (E7.5 and E8.5). Other chemokine receptor messages (CCR1, CCR4, CCR5, CCR2, and CXCR2) were found in E12.5 tissues concordant temporally and spatially with definitive (adult-like) hematopoiesis. Expression of CXCR4 was compared with that of its only known ligand, stromal cell derived factor-1 (SDF-1), by in situ hybridization. During organogenesis, these genes have dynamic and complementary expression patterns particularly in the developing neuronal, cardiac, vascular, hematopoietic, and craniofacial systems. Defects in the first four of these systems have been reported in CXCR4- and SDF-1 deficient mice. Our studies suggest new potential mechanisms for some of these defects as well as additional roles beyond the scope of the reported abnormalities. Earlier in development, expression of these genes correlates with migration during gastrulation. Migrating cells (mesoderm and definitive endoderm) contain CXCR4 message while embryonic ectoderm cells express SDF-1. Functional SDF-1 signaling in midgastrula cells as well as E12.5 hematopoietic progenitors was demonstrated by migration assays. Migration occurred with an optimum dose similar to that found for adult hematopoietic cells and was dependent on the presence of SDF-1 in a gradient. This work suggests roles for chemokine signaling in multiple embryogenic events. PMID- 10479461 TI - Health implications of DHA. PMID- 10479462 TI - Health benefits of docosahexaenoic acid (DHA) PMID- 10479463 TI - Is it really time to perform large intervention trials with antioxidant vitamins to prevent cardiovascular disease? PMID- 10479464 TI - Antioxidant vitamins and cardiovascular disease. PMID- 10479465 TI - Health benefits of docosahexaenoic acid (DHA) AB - Docosahexaenoic acid (DHA) is essential for the growth and functional development of the brain in infants. DHA is also required for maintenance of normal brain function in adults. The inclusion of plentiful DHA in the diet improves learning ability, whereas deficiencies of DHA are associated with deficits in learning. DHA is taken up by the brain in preference to other fatty acids. The turnover of DHA in the brain is very fast, more so than is generally realized. The visual acuity of healthy, full-term, formula-fed infants is increased when their formula includes DHA. During the last 50 years, many infants have been fed formula diets lacking DHA and other omega-3 fatty acids. DHA deficiencies are associated with foetal alcohol syndrome, attention deficit hyperactivity disorder, cystic fibrosis, phenylketonuria, unipolar depression, aggressive hostility, and adrenoleukodystrophy. Decreases in DHA in the brain are associated with cognitive decline during aging and with onset of sporadic Alzheimer disease. The leading cause of death in western nations is cardiovascular disease. Epidemiological studies have shown a strong correlation between fish consumption and reduction in sudden death from myocardial infarction. The reduction is approximately 50% with 200 mg day(-1)of DHA from fish. DHA is the active component in fish. Not only does fish oil reduce triglycerides in the blood and decrease thrombosis, but it also prevents cardiac arrhythmias. The association of DHA deficiency with depression is the reason for the robust positive correlation between depression and myocardial infarction. Patients with cardiovascular disease or Type II diabetes are often advised to adopt a low-fat diet with a high proportion of carbohydrate. A study with women shows that this type of diet increases plasma triglycerides and the severity of Type II diabetes and coronary heart disease. DHA is present in fatty fish (salmon, tuna, mackerel) and mother's milk. DHA is present at low levels in meat and eggs, but is not usually present in infant formulas. EPA, another long-chain n-3 fatty acid, is also present in fatty fish. The shorter chain n-3 fatty acid, alpha-linolenic acid, is not converted very well to DHA in man. These longchain n-3 fatty acids (also known as omega-3 fatty acids) are now becoming available in some foods, especially infant formula and eggs in Europe and Japan. Fish oil decreases the proliferation of tumour cells, whereas arachidonic acid, a longchain n-6 fatty acid, increases their proliferation. These opposite effects are also seen with inflammation, particularly with rheumatoid arthritis, and with asthma. DHA has a positive effect on diseases such as hypertension, arthritis, atherosclerosis, depression, adult-onset diabetes mellitus, myocardial infarction, thrombosis, and some cancers. PMID- 10479466 TI - Antioxidant vitamins and prevention of cardiovascular disease: laboratory, epidemiological and clinical trial data. AB - Naturally occurring antioxidants like vitamin E, beta-carotene, and vitamin C can inhibit the oxidative modification of low-density lipoproteins. This action could positively influence the atherosclerotic process and, as a consequence, the progression of coronary heart disease. A wealth of experimental studies provide a sound biological rationale for the mechanisms of action of antioxidants, whereas epidemiological studies strongly sustain the 'antioxidant hypothesis'. To data, however, clinical trials with beta-carotene supplements have been disappointing and their use as a preventive intervention for cancer and coronary heart disease should be discouraged. Only scant data from clinical trials are available for vitamin C. As for vitamin E, discrepant results have been obtained by the Alpha Tocopherol, Beta-Carotene Cancer Prevention Study with a low-dose vitamin E supplementation (50 mg daily) and the Cambridge Heart Antioxidant Study (400-800 mg daily). Currently ongoing are several large-scale clinical trials that will help in clarifying the role of vitamin E in the prevention of atherosclerotic coronary disease. PMID- 10479467 TI - Metal chelators/antioxidants: approaches to protect erythrocytic oxidative stress injury during Plasmodium berghei infection in Mastomys coucha. AB - Desferal, N-acetyl penicillamine (metal chelators) and propylgallate, catechin, and reduced glutathione (antioxidants) suppressed the erythrocytic oxidative damage generated during Plasmodium berghei infection in Mastomys coucha. Superoxide anion and lipid peroxide levels were increased and on the contrary, superoxide dismutase activity was noticeably decreased in the infected erythrocytes. Metal chelators/antioxidant treatment to infected animals resulted in restoration of O(2)(-), LPO and SOD to near normal levels. Furthermore, treatment of the above mentioned agents displayed a controlled mortality/survival time and parasitaemia. In conclusion, metal chelators/antioxidants were found to be effective against oxidative stress injury and parasite growth resulting in prolonged survival time of the host during experimental malaria. PMID- 10479468 TI - Anti-ulcer activity of SKP-450, a novel potassium channel activator, in rats. AB - The anti-ulcer effects of SKP-450, a new potassium channel activator, were evaluated on basal and histamine-induced gastric acid secretion, and against experimentally-induced ulcers such as ethanol-induced and NaOH-induced gastric ulcers. In the pylorus-ligated rat, SKP-450 (0.1-0.5 mg kg(-1)) significantly decreased volume and concentration of gastric juice, and total acid output (ED(50): 0.12 mg kg(-1)). SKP-450 (0.3-3.0 mg kg(-1)) also inhibited histamine induced gastric acid secretion, maximal effects being achieved at 1.0 mg kg( 1)(37.9% inhibition). In the 95% ethanol-treated rats, SKP-450 significantly reduced the mucosal lesions (46.9 and 31.4% inhibition at 0.1 and 0.2 mg kg(-1), respectively). A significant reduction in the ulcer index by SKP-450 was also observed in 0.3 n NaOH-treated rats (31.5 and 64.3% inhibition at 0.5 and 1.0 mg kg(-1), respectively). The effects of SKP-450 on histamine-induced acid secretion and on NaOH-induced ulcers were inhibited by glibenclamide (20 mg kg(-1), i.v.), a selective blocker of ATP-sensitive potassium channel. These results indicate that SKP-450 possesses anti-ulcer effects and its effects may be mediated by activation of ATP-sensitive potassium channels. PMID- 10479469 TI - Effects of nicergoline on the retinal and cortical electrophysiological responses in glaucoma patients: a preliminary open study. AB - PURPOSE: The retinal dysfunction and the delayed visual cortex responses shown by patients affected by glaucoma can be objectively assessed by Pattern Electroretinogram (PERG) and Visual Evoked Potentials (VEP) recordings. The present study aims to evaluate the effects of nicergoline on the retinal function and on the visual cortical responses in glaucoma patients. METHODS: Sixty patients (mean age 44.6+/-3.7) with open angle glaucoma were enrolled. The patients were divided into two groups: NG Group, where 30 patients were treated with nicergoline (Cebran((R)), 2 cps day) for 30 days; and CG Group, where 30 patients were not treated. Simultaneous recordings of PERG and VEP were performed in NG patients at the baseline, at 30 days after treatment with nicergoline (day 30), and at 45 days from the end of the treatment (day 75). PERG and VEP were recorded in CG patients at the baseline and after 30 and 75 days. The visual stimulus for recording PERGs and VEPs was a checkerboard whose elements subtended a visual arc of 60' and 15' with a 70% contrast, and alternated at a frequency of 2 Hz. RESULTS: At the baseline none of the electrophysiological parameters observed in NG Group patients differed (P>0.05) from those of CG Group patients. At days 30 and 75, in CG Group patients the values of the PERG and VEP parameters were unmodified (P>0.05) with respect to the baseline. In NG Group patients, the 30-day treatment period with nicergoline induced a significant (P<0.01) improvement of the PERG and VEP parameters. At day 75 all the electrophysiological parameters of NG Group did not differ significantly (P>0.05) from those at the baseline. CONCLUSION: Treatment with nicergoline induces an improvement of the retinal function and of the visual cortical responses in patients affected by glaucoma. This effect disappears within 45 days after the suspension of the treatment. PMID- 10479470 TI - Neurochemical studies with the anticonvulsant felbamate in mouse brain. AB - Felbamate (FBM) is a relatively novel anticonvulsant agent which has been reported to exert its antiepileptic effects by blockade of the glycine recognition site on the N-methyl-D-aspartate subtype of glutamate receptor and potentiation at the gamma-aminobutyric acid (GABA) type A receptor. An increasing number of antiepileptic drugs have, however, additional, neurochemical actions on the GABA and glutamate systems which may contribute to their anticonvulsant activity. As a result, we have investigated the effects of FBM on several GABA- and glutamate-related neurochemical parameters in mouse brain. Adult male ICR mice were randomised into two groups and administered FBM (0-100 mg kg(-1)) intraperitoneally either as a single dose or twice daily for 5 days. Four hours after the final dose, animals were killed and their brains removed for analysis of GABA, glutamate and glutamine concentrations and activities of GABA transaminase and glutamic acid decarboxylase. Single and repeated doses of FBM were without effect on all of the parameters investigated. These results appear to exclude the possibility that FBM, in addition to its known effects on GABA and glutamate receptors, exerts its antiepileptic effects via an action on the GABA- and glutamate-related neurochemical parameters chosen for investigation. PMID- 10479471 TI - Chronic nicotine pretreatment protects the blood-brain barrier against nicotine induced seizures in the rat. AB - This study was designed to investigate the possible protective actions of nicotine on cerebrovascular permeability in convulsions during nicotine-induced seizures. We have measured the permeability changes in the blood-brain barrier (BBB) macroscopically and spectrophotometrically by using Evans blue dye. Specific gravity measurements were also performed to assess brain edema which develops after blood-brain barrier opening. The experiments were carried out on Wistar rats. Rats were divided into two groups. They received acutely a convulsive dose of nicotine 3, 5, 8 and 9 mg kg(-1)i.p. or pretreated with a low dose of nicotine (0.8 mg kg(-1)i. p.) for 21 days followed by the procedure mentioned in the first group. Acute nicotine injection induced a significant increase in blood pressure and Evans-blue passage, despite a decline in specific gravity values. Low doses of chronic nicotine administration markedly reduced both the leakage of dye, and brain water content. Chronic treatment with low doses of nicotine (0.8 mg kg(-1)day(-1)s. c.) lessened the intensity of tonic clonic seizures observed with a single dose of 3, 5, 8 or 9 mg kg(-1)nicotine. The data presented here demonstrate that nicotine pretreatment results in decreased sensitivity to nicotine-induced seizures in rats. PMID- 10479472 TI - Effects of early postnatal exposure to low concentrations of carbon monoxide on cognitive functions in rats. AB - Male Wistar rats were exposed to 75 and 150 ppm of carbon monoxide (CO) from day 1 after birth until postnatal day 10 and their cognitive functions were evaluated at 3 and 18 months of age. The results show that early postnatal exposure to CO does not affect the acquisition and reacquisition of an active avoidance task in both adult and aged rats. Conversely, our previous findings indicate that prenatal exposure to CO (75 and 150 ppm), resulting in maternal blood carboxyhaemoglobin concentrations equivalent to those found in human cigarette smokers, induces long-lasting learning and memory deficits. These findings suggest that neurofunctional sequelae of prenatal CO exposure are notably different from those occurring in response to early postnatal exposure and that the vulnerability of the developing brain to prolonged, relatively mild, decrease in oxygen availability induced by CO critically depends on the particular period of developmental exposure. PMID- 10479473 TI - Inhibition by levamisole of the organic cation transporter rOCT1 in cultured rat hepatocytes. AB - Levamisole is known to be subject to hepatic removal and metabolism and to biliary excretion. The aim of our work was to study the mechanism involved in the removal of this compound by the liver. For this purpose, we studied the influence of levamisole on the uptake and efflux of the model organic cation 1-methyl-4 phenylpyridinium (MPP(+)) by primary cultured rat hepatocytes. Levamisole (500 microm) was found to produce a strong inhibition (to 31+/-2% of control) of [(3)H]MPP(+)uptake. Moreover, efflux of [(3)H]MPP(+)was also potently reduced by levamisole (500 microm). Our results show that levamisole interferes with an hepatic organic cation transporter which accepts MPP(+)as a substrate. This mechanism most probably corresponds to rOCT1, and it might be responsible for the hepatic removal of levamisole from the blood circulation. PMID- 10479474 TI - Pharmacokinetics of doxorubicin incorporated in solid lipid nanospheres (SLN). AB - The pharmacokinetics of doxorubicin incorporated as ion-pair into solid lipid nanospheres (SLN) was compared with that of the commercial solution of the drug. Male albino rats (Wistar-derived strain) were treated i.v. with equivalent doses (6 mg kg(-1)) of two different doxorubicin formulations: an aqueous dispersion of SLN carrying doxorubicin and a commercial doxorubicin solution (Adriablastina). These formulations were injected, under general anaesthesia, through a cannula into the jugular vein and blood samples were collected at 1, 15, 30, 45, 60, 120 and 180 min after administration. After 180 min rats were killed and samples of liver, heart, lung, kidney, spleen and brain were collected. Blood and tissue samples were analysed by a spectrofluorimetric method. The anthracycline concentration in the blood was markedly higher at each point times with the SLN than with the commercial solution. The drug concentration was also higher in the lung, spleen and brain. SLN-treated rats showed a lower doxorubicin concentration in liver, heart and kidney. The results showed that SLN increased the area under the curve (0-180 min) of doxorubicin compared to conventional doxorubicin solution and led to a different body distribution profile. PMID- 10479475 TI - Pharmacosurveillance in hospitalized patients in Italy. Study design of the 'Gruppo Italiano di Farmacovigilanza nell'Anziano' (GIFA). AB - The Italian Group of Pharmacoepidemiology in the Elderly (Gruppo Italiano di Farmacovigilanza nell'Anziano, GIFA) is a collaborative pharmacosurveillance study in hospitalized patients, sponsored by the Italian National Research Council (CNR) and the Italian Society of Gerontology and Geriatrics. It was founded in 1987 with the aim to constitute a multicentre research group to study quality of care and problems related to pharmacological therapy in the elderly. Until now the GIFA study has completed seven periodical surveys and enrolled a total of 28,411 hospitalized patients in 83 clinical centres. The database of the study contains approximately 174,000 in-hospital drug prescriptions, approximately 88,000 discharge diagnoses and a great deal of data on topical geriatric items such as cognitive performance, disability, comorbidity, adverse drug reactions and incontinence. This paper describes the general organization and the methods of the GIFA study and shows in detail the type of data collected. PMID- 10479476 TI - Effect of the meglitinide analog S21403 on cationic fluxes and insulin release in perifused rat pancreatic islets exposed to a high concentration of D-glucose. AB - The effect of the meglitinide analog S21403 (10 microm) upon(86)Rb and(45)Ca outflow and insulin release was investigated in perifused rat islets exposed to a high concentration of D-glucose (16.7 mm) in order to simulate the situation found in diabetic patients. Under these conditions, S21403 provoked a rapid, sustained and rapidly reversible increase in(86)Rb outflow, (45)Ca efflux and insulin release. These effects were suppressed or reversed when the experiments were conducted in the absence of extracellular Ca2+. They support the view that S21043 could be used as a novel insulinotropic tool in the treatment of non insulin-dependent diabetes mellitus, the cationic and secretory responses to the drug displaying a favourable time course for prompt and not unduly prolonged activation of islet B-cells. PMID- 10479477 TI - Niemann-Pick C1 is a late endosome-resident protein that transiently associates with lysosomes and the trans-Golgi network. AB - Niemann-Pick type C (NPC) disease is a severe cell lipidosis characterized by the accumulation of unesterified cholesterol in the endosomal/lysosomal system. Recently the primary disease-causing gene, NPC1, was identified, but few clues regarding its potential function(s) could be derived from its predicted amino acid sequence. Therefore, efforts were directed at characterizing the subcellular location of the NPC1 protein. Initial studies with a FLAG-tagged NPC1 cDNA demonstrated that NPC1 is a glycoprotein that associates with the membranes of a population of cytoplasmic vesicles. Immunofluorescence microscopy using anti-NPC1 polyclonal antibodies confirmed this analysis. Double-label immunofluorescence microscopy and subcellular fractionation studies indicated that NPC1 associates predominantly with late endosomes (Rab9 GTPase-positive vesicles) and, to a lesser extent, with lysosomes and the trans-Golgi network. When cholesterol egress from lysosomes was blocked by treatment of cells with U18666A, the NPC1 location shifted from late endosomes to the trans-Golgi network and lysosomes. Subcellular fractionation of liver homogenates from U18666A-treated mice confirmed these observations. These data suggest that U18666A may inhibit the retrograde transport of NPC1 from lysosomes to late endosomes for subsequent transfer to the trans-Golgi network. PMID- 10479479 TI - Trimethylaminuria is caused by mutations of the FMO3 gene in a North American cohort. AB - Trimethylaminuria (TMAuria) (McKusick 602079) first described in 1970 is an autosomal recessive condition caused by a partial or total incapacity to catalyze the N-oxygenation of the odorous compound trimethylamine (TMA). The result is a severe body odor and associated psychosocial conditions. This inborn error of metabolism, previously thought to be rare, is now being increasingly detected in severe and milder presentations. Mutations of a phase 1 detoxicating gene, flavin containing monooxygenase 3 (FMO3), have been shown to cause TMAuria. Herein we describe a cohort of individuals ascertained in North America with severe TMAuria, defined by a reduction of TMA oxidation below 50% of normal with genotype-phenotype correlations. We detected four new FMO3 mutations; two were missense (A52T and R387L), one was nonsense (E314X). The fourth allele is apparently composed of two relatively common polymorphisms (K158-G308) found in the general population. On the basis of this study we conclude that one common mutation and an increasing number of private mutations in individuals of different ethnic origins cause TMAuria in this cohort. PMID- 10479478 TI - Analyses of proteins involved in vesicular trafficking in platelets of mouse models of Hermansky Pudlak syndrome. AB - Hermansky Pudlak syndrome (HPS) is an autosomal recessive inherited disorder characterized by defects in synthesis and/or secretion of three related subcellular organelles: melanosomes, platelet-dense granules, and lysosomes. In the mouse, mutant forms of any of 14 separate genes result in an HPS-like phenotype. The mouse pearl and mocha genes encode subunits of the AP3 adaptor protein complex, confirming that HPS mutations involve proteins regulating intracellular vesicular trafficking. Therefore, expression of several additional proteins involved in vesicular transport was examined by immunoblotting of platelet extracts from HPS mutant and control mice. Platelet levels of SCAMPS (secretory carrier membrane proteins), Rab11, Rab31, NSF (N-ethylmaleimide sensitive fusion protein), syntaxin 2, syntaxin 4, munc18c, and p115/TAP (p115/transcytosis-associated protein) were not significantly altered in several different HPS mutants. However, gunmetal (gm/gm) platelets contained decreased amounts of SNAP-23. The Snap23 gene was mapped to mouse chromosome 5, demonstrating it cannot encode the gm gene, which maps to chromosome 14. It is likely therefore that the gm gene functions upstream of SNAP-23 in vesicular trafficking. PMID- 10479480 TI - Human liver-specific very-long-chain acyl-coenzyme A synthetase: cDNA cloning and characterization of a second enzymatically active protein. AB - Activation of fatty acids, catalyzed by acyl-coenzyme A (acyl-CoA) synthetases, is required for their subsequent metabolism. Peroxisomes and microsomes contain very-long-chain acyl-CoA synthetases (VLCSs) capable of activating fatty acids with a chain length of 22 or more carbons. Decreased peroxisomal VLCS activity is, in part, responsible for the biochemical pathology in X-linked adrenoleukodystrophy (X-ALD), illustrating the importance of VLCSs in cellular fatty acid homeostasis. We previously cloned two human genes encoding proteins homologous to rat peroxisomal VLCS; one (hVLCS) is the human ortholog to the rat VLCS gene and another (hVLCS-H1) encodes a related heart-specific protein. Here, we report the cloning of a third gene (hVLCS-H2) and characterization of its protein product. The hVLCS-H2 gene is located on human chromosome 19 and encodes a 690-amino-acid protein. The amino acid sequence of hVLCS-H2 is 44-45% identical and 67-69% similar to those of both hVLCS and hVLCS-H1. COS-1 cells transiently overexpressing hVLCS-H2 activated the very-long-chain fatty acid lignocerate (C24:0) at a rate >1.5-fold higher than that of nontransfected cells (P < 0.002). The hVLCS-H2-dependent activation of long- and branched-chain fatty acids following transient transfection was less striking. However, hVLCS-H2-dependent acyl-CoA synthetase activity with long- and very-long-chain fatty acid substrates was detected in COS-1 cells stably expressing hVLCS-H2. For all substrates tested (C18:0, C20:0, C24:0, C26:0), the hVLCS-H2 catalyzed activity was significantly increased (P < 0.01 to P < 0.0001). By both Northern analysis and reverse transcription polymerase chain reaction, hVLCS-H2 is expressed primarily in liver. Indirect immunofluorescence of COS-1 cells or human hepatoma-derived HepG2 cells expressing epitope-tagged hVLCS-H2 revealed that the protein was associated with the endoplasmic reticulum but not with peroxisomes. Thus, the primary role of hVLCS-H2 is likely to be in fatty acid elongation or complex lipid synthesis rather than in degradation. PMID- 10479482 TI - A type 2 diabetes-associated polymorphic ARE motif affecting expression of PPP1R3 is involved in RNA-protein interactions. AB - We have previously described a polymorphism in the 3' untranslated region (UTR) of the PPP1R3 gene that encodes the muscle-specific glycogen-targeting regulatory PP1 subunit. This polymorphism alters the distance between two putative mRNA destabilizing ATTTA (AUUUA) motifs and is distinguished by a 10-nucleotide (allele ARE1) vs a 2-nucleotide interval (allele ARE2). ARE2 is associated with insulin resistance as well as increased prevalence of type 2 diabetes in the Pima Indians, and correlates with reduced expression of this subunit in vivo, causing a 10-fold half-life reduction of reporter mRNA in NIH3T3 cells. Gel shift assays, Northwestern blotting, and RNA-protein UV crosslinking revealed three proteins (43, 80, and 139 kDa) binding to the polymorphic ARE region in these cells. The interactions are sequence specific, and can be suppressed by an unlabeled competitor in a dose-dependent manner. The less stable ARE2 allele shows at least 2-fold higher relative protein binding, indicating that the polymorphic ARE region has a mRNA-destabilizing role. We suggest that the increased protein binding to ARE2 contributes to a faster degradation of PPP1R3 mRNA carrying this allele, and the resulting lower concentration of the protein contributes to insulin resistance, thus increasing the risk for development of type 2 diabetes. PMID- 10479481 TI - The mutant genotype is the main determinant of the metabolic phenotype in phenylalanine hydroxylase deficiency. AB - Phenylketonuria and mild hyperphenylalaninemias are allelic disorders caused by mutations in the phenylalanine hydroxylase (PAH) gene. Following identification of the disease-causing mutation in 11 PAH-deficient patients, we tested the activity of the mutant gene products in an eukaryotic expression system. Two mutations markedly reduced PAH activity (A259V and L333F), one mutation mildly altered the enzyme activity (E390G), while the majority of mutant genotypes reduced the in vitro expression of PAH activity to 15-30% of controls. Comparing the predicted residual activity derived from expression studies to the clinical phenotypes of our PAH-deficient patients, we found that homozygosity for the L333F and E390G mutations resulted in severe and mild PAH deficiencies, respectively, both in vivo and in vitro, while compound heterozygosity (L333F/E390G) resulted in an intermediate dietary tolerance. Similarly, in vitro expression studies largely predicted dietary tolerance in compound heterozygotes for the A259V/IVS12nt1 (typical PKU), A259V/A403V, G218V/I65T, and G218V/R158Q mutations (mild variants). Taken together, these results support the view that expression studies are useful in predicting residual enzyme activity and that the mutant genotype at the PAH locus is the major determinant of metabolic phenotype in hyperphenylalaninemias. PMID- 10479483 TI - Determination of queuosine modification system deficiencies in cultured human cells. AB - Queuosine-deficient tRNAs are often observed in neoplastic cells. In order to determine possible sites for malfunction of the multistep queuosine modification system, comprehensive studies were performed on two human neoplastic cell lines, the HxGC(3) colon adenocarcinoma and the MCF-7 breast adenocarcinoma, which are 100 and 50-60% queuosine deficient, respectively. These results were compared with data obtained from normal human fibroblast (HFF) cultures which maintain 100% queuosine-modified tRNA populations. Queuine uptake in all three cell types was similar and each demonstrated activation by protein kinase C (PKC). However, incorporation of queuine into tRNA by tRNA:guanine ribosyltransferase (TGRase; E.C. 2.4.2.24) and PKC-catalyzed activation of this enzyme occurred only in HFF and MCF-7 cells. The HxGC(3) cell line exhibited no TGRase activity as was expected. Treatment with 5-azacytidine (5-azaC) induced TGRase activity to a level 20% of that in HFF and MCF-7 cells; however, this 5-azaC-induced TGRase activity was not regulated by PKC. Salvage of the queuine base from tRNA degradation products has been shown in mammalian cells and was measured in the HFF cells. However, salvage activity in the MCF-7 cell line was deficient. Therefore, it was shown by direct measurements that the HxGC(3) cell line is completely lacking in queuosine-modified tRNA due to loss of functional TGRase, while the MCF-7 cell line has an inefficient queuine salvage mechanism resulting in a significant deficiency of queuosine-modified tRNA. These techniques can be applied to any cultured cell types to determine specific lesions of the queuosine modification system, which have been suggested to be associated with neoplastic progression. PMID- 10479484 TI - Morphine regulation of a novel uridine diphosphate glucuronosyl-transferase in guinea pig pups following in utero exposure. AB - The uridine diphosphate glucuronosyltransferases (UGTs) catalyze conjugation reactions between various substrates and glucuronic acid, UDPGA (uridine diphosphate glucuronic acid), within the endoplasmic reticulum. Conjugation with UDPGA (glucuronidation) is an important pathway in the elimination, detoxification, and activation of compounds including steroid hormones, xenobiotics, and quaternary ammonium substrates. The guinea pig, which has a placental structure and a glucuronidation profile for morphine that are similar to the human, serves as a good small animal model to study the ontogeny of UGTs and the effect of in utero exposure to morphine on UGTs. We examined type 2 UGTs expressed in the guinea pig using amplification and cloning of partial cDNAs from liver RNA. Sequence analysis revealed a novel UGT2 (subsequently named UGT2A3),(2) that has a 64% amino acid sequence similarity to a known UGT2.(3) Full-length cDNAs were isolated from a guinea pig liver cDNA library. Tissue distribution of UGT2A3 using Northern blot analysis showed expression of three distinct size UGT2A3 mRNAs with unique expression in liver and small intestine. UGT2A3 mRNA is expressed at high levels in liver and lower levels in kidney and small intestine. In utero exposure to chronic intermittent morphine resulted in the up regulation of mRNA in 7-day-old female pups' liver and kidney as determined by quantitative RT-PCR analysis. The conjugation profile for UGT2A3 using stable expression in CHO cells and thin-layer chromatography demonstrated active conjugation of phenolic substrates. Regulation of UGTs by in utero morphine exposure may play an important role in fetal development. PMID- 10479485 TI - Combined adenine phosphoribosyltransferase and N-acetylgalactosamine-6-sulfate sulfatase deficiency. AB - We describe a Czech patient with combined adenine phosphoribosyltransferase (APRT) deficiency (2,8-dihydroxyadenine urolithiasis) and N-acetylgalactosamine-6 sulfate sulfatase (GALNS) deficiency (mucopolysaccharidosis Type IVA, Morquio disease A). Adenine and its extremely insoluble derivative, 2,8-dihydroxyadenine, were identified in the urine, and APRT deficiency was confirmed in erythrocytes. There was excessive excretion of keratan sulfate in the urine, and GALNS deficiency was confirmed in leukocytes. GALNS and APRT are both located on chromosome 16q24.3, suggesting that the patient had a deletion involving both genes. PCR amplification of genomic DNA indicated that a novel junction was created by the fusion of sequences distal to GALNS exon 2 and proximal to APRT exon 3, and that the size of the deleted region was approximately 100 kb. The deletion breakpoints were localized within GALNS intron 2 and APRT intron 2. Several other genes, including the alpha subunit of cytochrome B (CYBA), which is deleted or mutated in the autosomal form of chronic granulomatous disease, are located in the 16q24.3 region, but PCR amplification showed that this gene was present in the proband. A patient with hemizygosity for GALNS deficiency and APRT deficiency has been reported from Japan recently. These findings indicate that: (i) APRT is located telomeric to GALNS; (ii) GALNS and APRT are transcribed in the same orientation (centromeric to telomeric); and (iii) combined APRT/GALNS deficiency may be more common than hitherto realized. PMID- 10479487 TI - Linkage analysis of the hph-1 mutation and the GTP cyclohydrolase I structural gene. AB - Our research establishes genetic linkage between the hph-1 mutation and the GTP CH I structural gene. Our results indicate that these two loci are within an 8 cM region with 95% confidence. This finding lends additional support for the use of the hph-1 mouse mutant as a bona fide model system for the human disorder GTP-CH I to further our understanding of the molecular mechanisms involved in the disease pathology of GTP-CH I deficiency. PMID- 10479486 TI - Metabolic and secretory response to D-fructose in pancreatic islets from adult rats injected with streptozotocin during the neonatal period. AB - The metabolic and secretory responses to D-glucose and/or D-fructose were measured in pancreatic islets prepared from either control rats or animals that had been injected with streptozotocin during the neonatal period (STZ rats). The STZ rats displayed higher plasma D-glucose concentrations, but lower plasma insulin concentrations, islet insulin content, as well as basal and nutrient stimulated insulin release. This coincided with lower rates of D-[U-(14)C]hexose oxidation and D-[5-(3)H]hexose utilization. In both control and STZ rats, D fructose failed to affect significantly the metabolism of d-glucose, while the aldohexose increased the ratio between D-[U-(14)C]fructose oxidation and D-[5 (3)H]fructose conversion to (3)HOH. Such a ratio was higher than that found with radioactive D-glucose in islets exposed to both hexoses, whether in control or STZ rats, indicating a far-from-negligible contribution of fructokinase to the phosphorylation of D-fructose. Despite these analogies between both the respective fate of D-glucose and D-fructose and the reciprocal metabolic effects of the two hexoses in islets from control and STZ rats, the secretory response to the ketohexose in islets from STZ rats was preferentially suppressed, relative to that evoked by the aldohexose. This gives support to the idea that the insulinotropic action of D-fructose may not be entirely accounted for by its nutritional value in islet cells. PMID- 10479488 TI - A test for the future. PMID- 10479489 TI - A longitudinal study of psychological adjustment to familial genetic risk assessment for ovarian cancer. AB - OBJECTIVES: To evaluate the psychological adjustment of women during initial genetic ovarian cancer risk assessment and at clinic follow-up, 6-12 months later. METHODS: Sixty-five subjects were assessed with the Centre for Epidemiological Studies Depression Scale (CESD), Spielberger's State Anxiety Inventory, and an 18-item, investigator-designed questionnaire yielding self report on screening responses, worry about increased risk, identification of cancer-related deaths in relatives, worry about future cancer risks of daughters, alteration of future plans as a result of ovarian cancer risk, etc. RESULTS: Thirty-three percent of subjects had CESD scores above the established cutoff for depression at baseline and 38% had scores above cutoff at follow-up. Sixteen percent of subjects had state scores on the State-Trait Anxiety Inventory higher than 1 standard deviation above average (norm) at baseline, while only 6% had scores higher than 1 SD above average at follow-up. CONCLUSION: To identify factors associated with self-reported depression at follow-up, a series of demographic and self-reported variables (e.g., presence of identified problems in family, impact of genetic risk information, concern for daughter in the future) were entered in a multiple regression analysis with the CESD follow-up score as the dependent variable. Only one predictor accounted for a significant amount of variance in depression scores. Concern for daughter's risk in the future was associated with higher depression scores at follow-up (R = 0.33, P<0.02, R(2) = 11%). PMID- 10479490 TI - Analysis of telomerase activity in ovarian cystadenomas, low-malignant-potential tumors, and invasive carcinomas. AB - OBJECTIVE: Inappropriate telomerase expression has been reported to be associated with the development and/or progression of malignancies. Therefore, the purpose of the study was to determine and evaluate the levels of telomerase activity in normal ovary, cystadenomas, low-malignant-potential tumors, and carcinomas of the ovary. METHODS: In the present study, telomerase activity was examined in frozen tissue specimens of normal ovary (n = 6), ovarian cystadenomas (n = 13), ovarian low-malignant-potential (LMP) tumors (n = 12), and ovarian invasive carcinomas (n = 81). Clinicopathological information including age at diagnosis, histological grade, FIGO stage, presence of distant metastasis at diagnosis, and residual disease was available for all patients with ovarian carcinomas (n = 81). Telomerase activity was assessed by the telomeric repeat amplification protocol (TRAP). Arbitrary values were assigned to processivity of telomerase activities based on the molecular weights of the telomeric repeat DNA ladders, and were graded as "negative," "moderate" (< or =99 bp), or "high" (>100 bp) activities. The specificity of telomerase activity was determined by the pretreatment of telomerase-positive control or tumor samples with RNase that led to the abolition of the activity. In addition, to determine the possibility of false negativity due to the presence of telomerase inhibitors, TRAP assay was performed on each of the telomerase-negative specimens by mixing them individually with the telomerase positive control. RESULTS: Telomerase activity in the progression of ovarian carcinogenesis was evaluated. In comparison with normal ovary/cystadenoma (32%), a much higher frequency of the moderate activity was observed in LMP tumors (67%) or invasive carcinomas (57%), suggesting a close association between the latter two categories. The results reflect a subpopulation of telomerase-positive LMP tumor cells with the potential to develop invasive carcinomas. None of the specimens of the benign or LMP tumors exhibited high activity. In contrast, 18% of ovarian invasive carcinomas showed high telomerase activity (P = 0.013, Fisher exact test) and further 57%, moderate activity (75% in all). A statistically significant difference was observed in the expression of telomerase activity between normal ovary/benign cystadenomas and ovarian invasive carcinomas (P = 0.001, chi(2) test). CONCLUSIONS: The study showed a high prevalence of telomerase activity in LMP tumors or invasive carcinomas, the high levels of telomerase activity being associated exclusively with the invasive ovarian carcinomas. Therefore, the levels of processivity of telomerase activity and evidence of its statistically significant association with ovarian carcinoma suggest its role in the progression of ovarian carcinogenesis. PMID- 10479491 TI - Phase II trial of paclitaxel in leiomyosarcoma of the uterus: a gynecologic oncology group study. AB - OBJECTIVE: The aim of this study was to determine the activity of paclitaxel in chemotherapy-naive patients with advanced or recurrent uterine leiomyosarcoma. METHODS: Patients received 175 mg/m(2) of paclitaxel iv over 3 h. Courses were repeated every 3 weeks until disease progression or adverse side effects supervened. RESULTS: Thirty-four women were entered, but 1 patient was ineligible because of wrong cell type. Median age was 55 years (range: 35-84 years). GOG performance status was 2 in 2 instances, 1 in 9 cases, and 0 in 22 others. Eight patients (23.4%) had received radiotherapy. A median of 2.5 courses was given (range: 1-18). Eleven patients (33.3%) experienced grade 3 or 4 neutropenia, 1 (2.9%) had grade 4 thrombocytopenia, and 1 had grade 3 anemia. There were no cases of grade 3 or 4 gastrointestinal or dermatologic toxicity. One patient each developed deep venous thrombosis and a grade 3 allergic reaction. Eight patients (24.2%) had stable disease for at least 2 courses of therapy. Three complete responses were reported (9.1%). CONCLUSION: With the dose and schedule tested, paclitaxel has limited activity in patients with uterine leiomyosarcoma. Modest toxicity suggests that a higher dose of paclitaxel might be evaluated. PMID- 10479492 TI - SEER data, corpus uteri cancer: treatment trends versus survival for FIGO stage II, 1988-1994. AB - OBJECTIVE: 1998 Surveillance, Epidemiology, and End Results (SEER) data estimate an 83.1% 5-year survival rate for corpus uteri adenocarcinoma FIGO stage II. The SEER data were evaluated to determine whether primary treatment differences using simple hysterectomy or radical hysterectomy, with or without radiation, altered disease survival. MATERIALS AND METHODS: SEER incidence data for FIGO II uterine corpus cancer of adenocarcinoma histology from 1988 to 1994 were stratified by hysterectomy type (simple versus radical) and whether radiation was given. Survival rates were calculated using a relative survival method and are expressed as percentages. Statistical analysis was done using a Z test. RESULTS: The 5-year cumulative survival rate for patients with stage II uterine corpus adenocarcinoma who received surgery alone as primary therapy was 84.36% with simple hysterectomy and 92.96% with radical hysterectomy (P<0.05). Survival for patients who received combination radiation and surgery as primary therapy was 82.77% with simple hysterectomy and 88.02% with radical hysterectomy (P<0.05). Pelvic and para aortic nodes were negative. There was no significant survival difference for radiation versus no radiation in either surgical group. CONCLUSION: Radical hysterectomy is associated with better survival when compared to simple hysterectomy for FIGO II corpus uteri adenocarcinoma. PMID- 10479493 TI - Prognostic significance of DNA ploidy in endometrial cancer. AB - OBJECTIVE: The aim of this study was to analyze the prognostic significance of DNA ploidy in patients with endometrial cancer. METHODS: Between October 1988 and January 1997, DNA ploidy was determined prospectively in 208 women who were staged surgically by a standard protocol that included pelvic and para-aortic lymphadenectomy. Median follow-up was 48 months. RESULTS: Diploid tumors were identified in 154 (74%) patients and aneuploid tumors in 54 (26%). Patients with aneuploid tumors had a significantly higher prevalence of metastases to the cervix, adnexa, and omentum, malignant pelvic cytology, and advanced surgical stage. Patients with aneuploid tumors had a 4.5 times higher prevalence of pelvic lymph node metastases and a 5.8 times higher prevalence of para-aortic lymph node metastases. A significantly higher proportion of patients with aneuploid tumors was diagnosed with recurrent or progressive endometrial cancer (22.2 versus 6.5%, P = 0.002). Patients with aneuploid tumors had a significantly lower rate of survival from cancer death (P = 0.038) with 83% versus 94% surviving 5 years. CONCLUSION: Patients with aneuploid tumors are at high risk for lymph node metastases and should be surgically staged, including pelvic and para-aortic lymphadenectomy. Aneuploidy confers a risk for endometrial cancer death and these patients should be candidates for clinical trials evaluating treatment following surgery. PMID- 10479494 TI - Allelic losses at chromosome 3p are seen in human papilloma virus 16 associated transitional cell carcinoma of the cervix. AB - OBJECTIVE: Transitional cell carcinomas (TCCs) of the cervix are rare neoplasms of the female genital tract. Although these tumors display urothelial differentiation, there is controversy regarding their histogenetic relationship to squamous cell carcinomas (SCC) of the cervix versus transitional cell carcinomas of the bladder. METHODS: We performed partial allelotyping of five TCCs of the cervix using 23 polymorphic markers located on chromosomes 3p and 9, which demonstrate frequent and early losses in cervical SCC and urothelial TCC, respectively. Multiplex polymerase chain reaction was used on DNA extracted from archival paraffin-embedded tissue using precise microdissection. Additionally, P53 gene mutation analysis was performed using single-strand confirmation polymorphism (SSCP) and the presence of human papilloma virus (HPV) sequences was analyzed using general and specific (types 16 and 18) primers. RESULTS: General HPV sequences were demonstrated in all cases, but the oncogenic strain HPV 16 was present in only three (60%) of the five tumors; no HPV 18 was detected in any sample. Three of five TCCs, all harboring HPV 16 sequences, demonstrated concurrent allelic losses at several 3p loci (specifically 3p12, 3p14.2 [the FHIT gene locus], 3p21.3, and 3p22-24.2). LOH at a single locus on 9q32-qter was demonstrated in one tumor; no other deletions were seen on chromosome 9. P53 gene mutations in exons 5-8 were absent by SSCP analysis. CONCLUSIONS: The infrequent involvement of chromosome 9 in TCCs of the cervix, along with the concurrent presence of 3p LOH and oncogenic HPV 16 in a subset of tumors, suggests a closer histogenetic relationship of this neoplasm to cervical SCCs rather than urothelial TCCs. PMID- 10479495 TI - Prognostic significance of epithelial-stromal vascular cuffing and microvessel density in squamous cell carcinoma of the uterine cervix. AB - OBJECTIVE: Tumor angiogenesis has been shown to play an important role in tumor growth and metastasis. This study examines the prognostic significance of two histological markers of angiogenesis, i.e., vascular cuffing (VC), a bead-like arrangement of microvessels closely surrounding microscopic tumor nests, and microvessel density (MVD), the number of microvessels in a unit area, in cervical squamous cell carcinoma. METHODS: One hundred twenty-two specimens from surgically resected uteri with cervical squamous cell carcinoma were histologically reviewed and immunostained for CD34. VC was graded into "none," "incomplete," and "complete." The MVD was determined by counting the microvessels with a light microscope within a x200 field area where neovascularization occurred most actively. Stromal inflammation was also split into three grades. The relationship of VC or MVD to clinicopathological prognostic factors such as FIGO stage, cervical stromal invasion, lymph-vascular space invasion, pelvic lymph node metastasis, and parametrial invasion was evaluated using univariate and multivariate analyses. RESULTS: The patients with a complete VC pattern showed a significantly worse prognosis compared to those with a pattern graded as either none or incomplete (P<0.011 and P<0.0001, respectively). The Cox regression analysis revealed the complete VC pattern, together with parametrial invasion, to be an independent prognostic indicator for overall survival. MVD and the grading of stromal inflammation showed no significant relationship with VC or overall survival. CONCLUSIONS: The complete VC pattern may therefore be a useful prognostic indicator in cervical squamous cell carcinoma. PMID- 10479496 TI - Expression and imprinting of insulin-like growth factor II (IGF2) and H19 genes in uterine leiomyomas. AB - Genomic imprinting is defined as a gamete of origin-specific epigenetic modification of DNA leading to differential gene expression in the zygote. Several imprinted genes have been identified and some of them are associated with tumor development. We investigated the expression and the imprinting status of IGF2 and H19 genes in 47 uterine leiomyomas. Using allelic transcription assay, we detected the expression of the IGF2 gene in 10 of a total of 15 informative cases. No loss of imprinting, as determined by the finding of biallelic expression, was detected in any case. The expression of H19 gene was detected in 10 of 20 informative cases and the imprinting pattern was also maintained in all of them. Our data suggest that alterations in IGF2 and H19 genes expression by loss of imprinting do not occur in uterine leiomyomas. PMID- 10479497 TI - Reporting normal endometrial cells in Pap smears: an outcome appraisal. AB - OBJECTIVE: The purpose of this study was to determine the clinical relevance of reporting the presence of normal endometrial cells in the Pap smears of women over the age of 35 years and the significance of this practice as it relates to patient management. METHODS: From January 1992 to December 1995, normal endometrial cells were reported in 206 consecutive Pap smears of women over the age of 35 years. Clinical follow-up was available for all patients, including the results of diagnostic procedures whenever performed. RESULTS: Of the 206 women with normal endometrial cells in their Pap smears, 162 presented with the chief complaint of abnormal vaginal bleeding. They were all evaluated by direct endometrial sampling, resulting in detection of 10 endometrial hyperplasias and 7 endometrial carcinomas. The remaining 44 women who were clinically asymptomatic were followed up with only routine annual gynecologic examinations for a minimum of 3 years. All had negative clinical courses. CONCLUSION: Reporting the presence of normal endometrial cells in Pap smears has little, if any, impact on subsequent patient management. Women who present with abnormal uterine bleeding are worked up for endometrial disease regardless of their Pap smear findings. In clinically asymptomatic patients, practitioners may, and in our experience often do, choose to disregard normal endometrial cells in Pap smear reports. The negative follow-up for the asymptomatic women in our study supports this practice. Therefore, reporting the presence of normal endometrial cells in Pap smears is of no clinical relevance and may, in fact, create a management dilemma for clinicians. PMID- 10479498 TI - Epidemiologic and surgicopathologic findings of papillary serous and clear cell endometrial cancers when compared to endometrioid carcinoma. AB - PURPOSE: The aim of this study was to identify similarities and differences in epidemiologic and surgicopathologic staging results for papillary serous (PS) and clear cell (CC) endometrial cancers compared with endometrioid (EM) carcinoma of the endometrium. METHODS: Clinical and surgicopathologic data were retrospectively collected on 574 clinical stage I-II endometrial cancer patients, including 53 PS and 18 CC (based on postoperative histology), undergoing hysterectomy at Duke University Medical Center between 1967 and 1990. All staging material was available and reexamined prior to this analysis, and FIGO surgical staging was retrospectively assigned. PS and CC histologic subtypes were compared both as a common category and as discrete categories versus EM, EM grade 1 (EM1), EM grade 2 (EM2), and EM grade 3 (EM3). Fisher's exact test was used to compare proportions with unordered categories (2x2 tables), while the chi(2) test for trend was used to compare proportions in 3x2 tables with ordered categories. Differences in medians were compared with the Wilcoxon rank-sum test. RESULTS: PS tumors accounted for 8%, CC for 2%, and EM for 90% of cases. Overall, 14% of tumors were changed to a different postoperative histology including 64% of PS, 50% of CC, and 8% of EM. Postoperative histology changes were 4% for EM1 and 21% for EM3. PS, CC, and EM3 had more surgical sampling performed than for other EM. Rates for lymph node dissections were similar for EM3 (81%), PS (72%), and CC (67%) tumors, although metastases were more frequent for PS and CC compared with EM3. When PS tumors were confined to the endometrium, paraaortic metastases occurred in 13%. LVSI increased with EM grade and was highest for PS and CC. Upstaging to surgical stage III-IV occurred in 47% of PS, 39% of CC, and 12% of EM. The majority of PS and CC tumors were confined to the inner one-third of the myometrium, compared with EM tumors, where grade correlated with depth of myometrial invasion. Extrauterine metastases occurred in 55% of PS and 45% of CC tumors confined to the inner one-half, compared with 17% of EM3. CONCLUSION: Frequent changes from preoperative to postoperative histology and grade may contribute to misassignment of preoperative and intraoperative risk as determined by depth of myometrial invasion for PS and CC patients. The higher frequency of extrauterine metastases in PS and CC tumors compared with EM3, despite similar surgical sampling rates, supports a more virulent behavior. The poor correlation between depth of myometrial invasion and risk for extrauterine metastases helps to explain poorer survival in PS and CC patients, in addition to more frequent upstaging. These results support routine extended surgical staging for women with preoperative or intraoperative diagnosis of PS and CC tumors. Intraoperative assessment of tumor grade and histology may be indicated and warrants further investigation. PMID- 10479499 TI - Mutational analysis of the PMS2 gene in sporadic endometrial cancers with microsatellite instability. AB - OBJECTIVE: Approximately 20% of endometrial tumors have a defect in DNA mismatch repair and exhibit microsatellite instability (MSI). We assessed the role of the PMS2 DNA mismatch repair gene in MSI-positive sporadic endometrial tumors. METHODS: We examined 40 sporadic endometrial tumor specimens with MSI. All 15 exons of the PMS2 gene were investigated for sequence alterations by single strand conformational variant analysis. RESULTS: Twelve polymorphisms were identified, 8 of which were in the coding sequence. Four specimens revealed mutations in intronic sequences that are not predicted to affect the PMS2 mRNA. No mutations were detected within the coding region of the PMS2 gene. CONCLUSION: We conclude that structural mutations in the PMS2 gene are not responsible for defective DNA mismatch repair in sporadic endometrial cancers with MSI. The identification of single nucleotide polymorphisms in the PMS2 locus may aid in the mapping and characterization of genetic diseases. PMID- 10479500 TI - The prognostic significance of peritoneal seeding and size of postsurgical residual in patients with stage III epithelial ovarian cancer treated with surgery, chemotherapy, and high-dose radiotherapy. AB - OBJECTIVE: The aim of this study was to retrospectively analyze the prognostic importance of age, histologic type and grade, ascites, lymph node status, size and type of postoperative residual disease, and radiation dose on disease specific (DSS) and progression-free survival (PFS) in stage III epithelial ovarian cancer patients who had been treated with radical surgery, postoperative chemotherapy, and high-dose radiotherapy. METHODS: Consolidation radiotherapy including whole abdominal radiation, pelvic, and upper abdominal boosts was employed in 46 patients who showed no evidence of residual or progressive disease after completion of multiagent chemotherapy. The median follow-up for all patients was 36 months and 103 months for patients at risk. The prognostic impact of pretreatment and treatment parameters on DSS and PFS was tested in univariate and multivariate analyses. RESULTS: The 5-year DSS and PFS rates for all patients were 38 and 33%, and for patients with 0-< or =2 cm residual tumor 65 and 61%, respectively. In univariate analysis, initial peritoneal seeding (both: P = 0.02), ascites (P = 0.03; 0.01), size of residual (0-< or =2 cm vs >2 cm), and residual miliary subdiaphragmatic (MDS) and localized peritoneal seeding (LPS) in the upper abdomen (P = 0.0002; 0.0003) were significantly correlated with DSS and PFS. Dose of radiation (< or =30 vs >30 Gy) correlated with DSS only (P = 0.02). In multivariate analysis size of residual disease (0-< or =2 cm vs >2 cm and/or MDS or LPS) remained the only independent prognostic factor for DSS and PFS (both; P = 0. 001). CONCLUSION: Patients with localized peritoneal seeding who were rendered free of disease elsewhere had an outcome equally poor as that of patients with gross residuals (>2 cm) in the upper abdomen. If our findings can be confirmed, attempted resection of all localized seeding in patients who are otherwise cytoreducible to no or minimal residual disease may be considered in combination with Taxol-containing regimens as are now being utilized for patients with gross disease. PMID- 10479501 TI - An alternative HER-2/neu transcript of 8 kb has an extended 3'UTR and displays increased stability in SKOV-3 ovarian carcinoma cells. AB - HER-2/neu is a potent oncogene that predicts poor outcome when overexpressed in ovarian cancer. The SKOV-3 ovarian carcinoma cell line, one of the only models for HER2-driven ovarian cancer, expresses a major uncharacterized 8-kb alternative HER-2 transcript. OBJECTIVES: The aim of this study was to characterize the structure and determine the origin of the alternative sequence and examine the possible role of the 8-kb alternative transcript in overexpression of the HER-2 gene. METHODS: The structure of the 8-kb transcript was investigated using polymerase chain reaction (PCR) and nucleotide sequencing of cDNA clones. PCR analysis of genomic DNA was used to assess the origin of the 8-kb transcript. The stability of the 8-kb mRNA was assessed by Northern blot analysis of RNA extracted from cells treated with transcriptional inhibitors. RESULTS: Similar 5'UTR and coding sequence but an extended 3'UTR were contained in the 8-kb compared to the well-characterized 4.5-kb HER-2 transcript. Genomic DNA had continuity between the novel 3'UTR sequence from the 8-kb transcript and adjacent HER-2 terminal exon sequence. The 8-kb transcript had a half-life of 13 h compared to 5.5 h for the 4.5-kb transcript (P<0.01). CONCLUSIONS: The 8-kb transcript is generated from alternative polyadenylation site usage rather than gene rearrangement. Since the 8-kb transcript contains alternative sequence found at the 3' end of the normal HER-2 gene, it could be expressed in other cells. Increased stability of the 8-kb transcript may confer a selective advantage for SKOV-3 cells by providing enhanced HER-2 expression. PMID- 10479502 TI - Interstitial brachytherapy for vaginal recurrences of endometrial carcinoma. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy of interstitial brachytherapy in the management of vaginal recurrences of endometrial carcinoma. METHODS: Thirty patients received interstitial irradiation, with or without external beam radiotherapy. They were followed for a minimum of 5 years or until death. RESULTS: The median age was 66 years at initial diagnosis of endometrial cancer. FIGO stages included Stage I (n = 18), Stage II (n = 7), and Stage III (n = 5). All patients were treated originally by total abdominal hysterectomy and bilateral salpingo-oophorectomy, with or without lymphadenectomy, and 13 (43%) also received postoperative adjuvant whole pelvis radiotherapy as part of their primary treatment. Vaginal recurrences were diagnosed at a mean interval of 29 months after hysterectomy (range, 3-119 months). No patient had clinical evidence of pelvic sidewall extension or of distant metastatic disease. All patients were treated with interstitial brachytherapy; each implant delivered a mean maximal tumor dose of 25.5 Gy. Eighteen patients (60%) also received external beam radiotherapy (mean dose, 48 Gy) as part of their treatment for vaginal recurrence. Twenty-eight patients (93%) experienced a complete clinical response. Ten patients relapsed in the vagina (n = 5) or at distant sites (n = 5). Eleven patients are dead of disease. From the time of vaginal recurrence, the median overall survival was 60 months and the cause of death adjusted 5-year survival rate was 65%. Major morbidity included radiation proctitis (n = 2), fistula (n = 2), and radiation stricture (n = 1). CONCLUSION: Interstitial irradiation resulted in favorable local control as well as a 5-year survival rate and morbidity comparable to that reported previously for conventional brachytherapy. PMID- 10479503 TI - Prognostic factors for relapse and pelvic lymph node metastases in early stage I adenocarcinoma of the cervix. AB - OBJECTIVES: The aim of this study was to assess the prognostic significance of local tumor factors in predicting lymph node metastases and/or recurrence in early adenocarcinoma of the cervix. METHODS: Patients were selected from the prospective computerized cervical cancer database of the division of gynecologic oncology. All patients had radical surgery and pelvic lymph node dissection. The study population consisted of all patients with stage I adenocarcinoma having tumor thickness <10 mm. Pathology was re-reviewed to assess histological subtype, depth, volume, grade, and presence of capillary lymphatic space involvement. RESULTS: The study group consisted of 68 patients, with a mean age of 40 years. The median follow-up was 40 months (range 8-102 months). The median tumor depth and volume were 2.8 mm (range 0.3-8.0 mm) and 237 mm(3) (range 0.1-7996 mm(3)), respectively. Twenty-two patients had tumor volumes greater than 600 mm(3), and of these, 5 (23%) patients either had positive pelvic lymph nodes (2) or developed recurrent disease (3) (none node positive) at a median time of 49 months. In comparison, 46 patients (68%) had tumor volumes of less than 600 mm(3), none of whom had positive pelvic lymph nodes or developed recurrence (P<0.005). Only 1 of 20 patients with a depth of invasion <2 mm had a tumor volume >600 mm(3) in comparison to 21 of 48 patients with deeper invasion (P<0.002). CONCLUSION: The incidence of positive pelvic lymph nodes and/or recurrence in this patient population is very low. As all patients with metastatic disease or recurrence had tumor volumes >600 mm(3), volume of disease rather than depth of invasion may be the single most important prognostic factor for the above events. However, many more patients will have to be studied to confirm this. PMID- 10479504 TI - The efficacy of cervical conization in the treatment of cervical intraepithelial neoplasia in HIV-positive women. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy of cervical conization in the treatment of CIN in HIV-positive women. MATERIALS AND METHODS: Sixty-six HIV-positive women treated with cervical conization for CIN were stratified into four groups based on surgical margin and endocervical curetting (ECC) status (group 1: -margin/-ECC, group 2: +margin/-ECC, group 3: +margin/+ECC, group 4: -margin/+ECC). The rate of histologically proven recurrent CIN was calculated for each group and compared using chi(2) analysis. The effect of +margins, +ECC, degree of dysplasia, and CD4 count on the risk of recurrence was determined by logistic regression. RESULTS: Forty-nine percent of patients with negative margins and negative ECC experienced recurrence, most within 36 months. There was no significant difference in recurrence rate for patients with positive margins (69.2%, P = 0.19), positive ECC (50%, P = 0.97), or positive margins and ECC (66.7%, P = 0.41) when compared to patients with complete excision of dysplasia. No significant difference in the mean CD4 count of patients with and without recurrent dysplasia (316 vs. 390 cells/mm3, P = 0.37) was observed. Logistic regression showed only degree of dysplasia in the cone specimen to have a marginally significant linear relationship with recurrence. CONCLUSION: Cervical conization is not an effective method for eradicating CIN in HIV-positive women. Most patients will recur despite complete excision of dysplasia. Surgical margin status, ECC status, and CD4 count appear to have no effect on recurrence rate. Although multiple procedures were necessary in some patients, cone biopsy was effective in preventing progression to invasive cervical cancer in all cases. PMID- 10479505 TI - Phase II trial of oral etoposide in recurrent or refractory endometrial adenocarcinoma: a southwest oncology group study. AB - Oral etoposide has activity in a wide variety of tumors and is well tolerated. Therefore, the efficacy of oral etoposide was assessed as a treatment of metastatic endometrial cancer. To be eligible for this group-wide Southwest Oncology Group trial, patients had to have histologically proven metastatic or recurrent endometrial carcinoma; no previous cytotoxic therapy; and adequate renal, hepatic, and hematologic function, and they had to have given informed consent. Therapy consisted of oral etoposide, 50 mg daily on days 1-21 on a 28 day schedule. Therapy was continued in the absence of toxicity or disease progression. Forty-four eligible women, with a median age of 68 years (range 38 84 years) were treated. Radiotherapy had been delivered to 33 and hormomal therapy to 21. The median duration of therapy was 69 days (range 7-510 days). The treatment was well tolerated. Only one patient had grade 4 neutropenia, and a second had grade 4 anemia. Three patients had grade 3 nausea. One complete and five partial responses (14%) were observed. An additional four patients had unconfirmed responses. Tumor regressions were noted in nodes, bone, and visceral organs. While oral etoposide has only a modest level of activity when used in chemonaive patients, the minimal toxicity of this drug makes it a candidate for use in combination chemotherapy. PMID- 10479506 TI - Impact of surgical staging in women with locally advanced cervical cancer. AB - OBJECTIVE: The aim of this study was to evaluate the impact of surgical staging in the treatment and outcome of women with locally advanced cervical cancer. METHODS: Ninety-eight women with locally advanced cervical cancer treated between 1993 and 1997 were retrospectively reviewed. Survival probabilities were calculated by the Kaplan-Meier product limit method and compared with the log rank test. RESULTS: Of the 98 women treated over the 5-year period, 86 were surgically staged: 61 by a retroperitoneal approach, 18 by laparoscopy, and 7 by laparotomy. Median blood loss was 120 cc and median length of hospitalization was 3 days. Preoperative CT scans (n = 55), when compared with surgical findings, missed macroscopic nodal disease in 20% and microscopic disease in 15% and overcalled disease in 10% of cases. Lymph node metastases were found in 45/86 patients (52%): 12 microscopic and 33 macroscopic. The highest level of nodes found to be involved was pelvic in 23, common iliac nodes in 3, para-aortic nodes in 14, and scalene nodes in 5 cases. Of the 86 patients, 49 received pelvic radiation, 27 received extended field radiation, and 10 were identified for palliative treatment only (5 scalene node metastasis, 5 extensive intraperitoneal disease). For node-negative patients, 5-year survival was 74%; for microscopic nodal involvement it was 58%; and for macroscopic involvement it was 39% (P = 0.007). Five-year survival for women with para-aortic node involvement was 52%. Number of nodes involved was a significant prognostic variable (P = 0.008). Patients who received chemotherapy had a 5-year survival of 68% compared to 35% for those who did not (P = 0.06). Factors which did not affect survival included age, histology, type of surgery, stage, and type of radiation (pelvic vs extended). CONCLUSION: Surgical staging of women with locally advanced cervical cancer can be performed with acceptable morbidity and it provided more accurate information than CT scans and resulted in a modification of the standard pelvic radiation field for 43% of our patients. The information obtained from surgical staging allows better individualization of therapy, which may improve overall clinical outcome. PMID- 10479507 TI - A comparison of treatment strategies for endometrial adenocarcinoma: analysis of financial impact. AB - OBJECTIVE: The aim of this study was to investigate the influence of three treatment strategies for adenocarcinoma of the endometrium on the utilization of adjuvant radiation therapy and the medical charges associated with each pattern of practice. METHODS: Three clinical algorithms felt to represent practice patterns for patients with endometrial cancer were considered: (1) comprehensive surgical staging of all patients, with adjuvant pelvic radiation reserved for documented cases of extrauterine disease, (2) total abdominal hysterectomy with bilateral salpingo-oophorectomy (TAH/BSO) with lymph node dissection reserved for cases of myometrial invasion, followed by adjuvant radiation based on the presence of uterine risk factors, and (3) TAH/BSO followed by intraoperative pathologic assessment of the uterus and consultation with a "surgical" oncologist for comprehensive staging. Each algorithm was applied to a cohort of 190 surgically staged patients identified through a retrospective medical records review. The use of radiation in each algorithm was quantified and the associated financial impact was estimated using hospital charges. RESULTS: Treatment algorithm 1 yielded the lowest charges per patient at $12,778.52. Treatment algorithms 2 and 3 had associated charges per patient of $15,997.02 and $17,343.44, respectively. CONCLUSION: Approaches to care that lead to cost effective utilization of health care resources should be pursued. PMID- 10479508 TI - Generation of tumor-specific cytotoxic T lymphocytes by stimulation with HPV type 16 E7 peptide-pulsed dendritic cells: an approach to immunotherapy of cervical cancer. AB - OBJECTIVE: The aim of this study was to generate HPV-16 E7 peptide-specific cytotoxic T lymphocytes (CTLs) in vitro for future adoptive immunotherapy of cervical cancer. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from HLA-A2+ healthy donors. The PBMCs were incubated with HPV-16 E7(11-20) peptide and varying cytokines in the primary culture. Restimulation was performed weekly with peptide-pulsed, irradiated autologous PBMCs. Alternatively, the PBMCs were depleted of abundant CD4+ cells and stimulated with HPV-16 E7(11-20) peptide pulsed dendritic cells. Cytolytic activity was determined by a standard 4-h (51)Cr-release assay. RESULTS: After 6 weeks in culture, we were able to establish peptide-specific CTL lines in one of seven donors by incubating PBMCs with HPV-16 E7(11-20) peptide. When we employed autologous peptide-pulsed dendritic cells to stimulate CD8+ cell-enriched PBMCs, we obtained CTL lines in four of seven donors. The primed CTLs were able to lyse the HLA-A2+ and HPV-16+ cervical cancer cell line Caski. SiHa, an HLA-A2-, but HPV 16+, cervical cancer cell line could be lysed only after transfection with HLA-A2. In addition, a high cytotoxicity (>80%) was obtained against peptide-pulsed, but not unpulsed, targets such as autologous Ebstein-Barr virus-immortalized B cells or allogeneic lipopolysaccaride-stimulated PBMCs. DCs were clearly the most potent of all tested antigen presenting cells to stimulate a CTL response in a proliferation assay. CONCLUSION: HPV-16 E7 peptide-specific CTLs could be generated in vitro. A practical protocol to expand the CTLs to a sufficient number for an application in a clinical trial is in progress. PMID- 10479509 TI - "Curbside" consultations in gynecologic oncology: a closer look at a common practice. AB - OBJECTIVE: The aim of this article is to evaluate the nature and frequency of informal "curbside" consultations in a gynecologic oncology practice. METHODS: A log of all curbside consultations requested from a gynecologic oncologist in a tertiary multispecialty group practice was kept for 2 years (July 1996 to June 1998). New referrals or calls concerning mutual patients were excluded. RESULTS: Three hundred thirty-three requests for advice were received from 87 different physicians. Most (81%) of the calls were from 51 obstetrician/gynecologists (Ob/Gyns); 13% of the calls were from family practitioners or general internists. The four most common topics discussed were lower genital tract dysplasia (18%), endometrial hyperplasia (17%), adnexal masses (11%), and endometrial cancer (10%). Overall, 29% of questions pertained to the management of an invasive gynecologic malignancy. Twenty-four percent of the questions were answered in less than 5 min, 43% in 5-10 min, 29% in 11-20 min, and 4% in over 20 min. The issues discussed were considered simple in 26%, of moderate complexity in 49%, very complex in 22%, and ultracomplex in 3%. One hundred nine questions (33%) were of sufficient complexity to warrant referral to a specialist, which occurred in 52 cases (48%); physicians employed by a staff-model health maintenance organization were less likely to request a formal consultation (14%, chi(2) = 0.001). CONCLUSIONS: Curbside consultations are a frequent occurrence in the practice of a gynecologic oncologist, with most questions posed by general Ob/Gyns. Approximately one-third of the questions were of minimal complexity and involved topics covered in standard textbooks or residency training programs. In contrast, approximately one-third were of sufficient complexity that a casual discussion between colleagues may not yield optimal medical care. PMID- 10479510 TI - Effect of cervical disease history on outcomes of women who have a pap diagnosis of atypical glandular cells of undetermined significance. AB - OBJECTIVE: To determine the relationship between history of cervical dysplasia or carcinoma and the development of cervical dysplasia or adenocarcinoma in women who have a diagnosis of atypical glandular cells of undetermined significance (AGUS), favor endocervical origin, or AGUS, not otherwise specified. METHODS: A 6 year retrospective review of the pathology files was performed for 93 women who were diagnosed in 1992 with AGUS, favor endocervical origin, or AGUS, not otherwise specified. Data collected included previous history of cervical disease, follow-up diagnoses, time interval between follow-ups, and procedures performed. RESULTS: Of women with follow-up who had or did not have a previous history of cervical dysplasia, 32.0 and 12.0%, respectively, developed a squamous dysplasia or adenocarcinoma in situ. This difference was statistically significant (P<0.05). Of the women who had or did not have a previous history of cervical dysplasia and had Pap smear follow-up, only 4.2 and 4.3%, respectively, had a false-negative diagnosis on the most immediate subsequent smear. CONCLUSIONS: Women who have AGUS, favor endocervical origin, or AGUS, not otherwise specified, and no history of cervical dysplasia have a significantly lower risk of developing or having cervical dysplasia than women who have the same diagnosis and a history of cervical dysplasia. This may warrant different treatment protocols for these two groups. For the women with AGUS and no previous history of cervical dysplasia, a repeat Pap smear, rather than colposcopy with curettage, may be warranted. PMID- 10479511 TI - What staging surgery should be performed on patients with uterine papillary serous carcinoma? AB - OBJECTIVE: While uterine papillary serous carcinoma (UPSC) is an aggressive histologic subtype, it fortunately is not as common as some other histologic subtypes. Overall, patients with UPSC have a poor survival rate. Since the optimal surgical procedure to perform on patients with this tumor is unknown, the authors wanted to determine what the optimal surgical management of patients with UPSC should be. METHODS: All patients with the preoperative or frozen section intraoperative diagnosis of UPSC were treated with a staging or cytoreductive procedure analogous to patients with serous carcinoma of the ovary. Patients analyzed underwent surgery from March 1983 to September 1995. RESULTS: Sixty-five patients with UPSC were found. Twenty patients had FIGO stage I tumors, 6 stage II tumors, 8 stage III tumors, and 31 stage IV tumors. Twenty-nine patients had upper abdominal disease (17 gross disease and 12 microscopic disease only). Forty eight patients underwent pelvic and paraaortic lymphadenectomy, with 6 of 48 having positive lymph nodes. All 14 patients with lymphovascular space invasion had stage IV disease. Thirty-one of sixty-five patients had positive cytology at the time of surgery. CONCLUSION: Based on the clinical experience of these investigators, patients with UPSC should undergo a staging laparotomy similar to the procedure undertaken for patients with ovarian carcinoma. The surgery should include at least partial omentectomy, total abdominal hysterectomy and bilateral salpingo-oophorectomy, peritoneal washings, peritoneal biopsies, and pelvic and paraaortic lymphadenectomy similar to an ovarian cancer staging procedure if no gross disease > or =2 cm is found at time of surgery. If disease > or =2 cm is found, cytoreduction should be undertaken when feasible. PMID- 10479513 TI - Ovarian cancer in young women in Sweden, 1989-1991. AB - The present study is an evaluation of the symptoms and gynecological status, at the time of seeking medical advice, among women of reproductive age diagnosed with ovarian malignancies. The study was based on the medical records of all women in Sweden between 15 and 40 years of age who were diagnosed with ovarian malignancies during a 3-year period (1989-1991). The study focused on the diagnosis and panorama of symptoms. These data were compared to the records of more than 10,000 women in this age group during the same period of time who underwent surgery for ovarian changes that resulted in benign diagnoses. The 152 cases of ovarian cancer included only 1 patient who was without symptoms, had normal pelvic status upon manual examination, and a sonographically diagnosed simple cyst. These data support the conclusion that the majority of young women with ovarian cancer have symptoms and/or clinically detectable adnexal masses at the time when medical advice is sought. These studies suggest that diagnosis of ovarian cancer is unlikely in young healthy women lacking subjective symptoms and an adnexal mass upon pelvic examination. In conclusion, programs for the management of simple cysts and development of noninvasive diagnostic methods for ovarian changes found in patients in the reproductive age group are needed in order to reduce the number of patients subjected to operative intervention. PMID- 10479512 TI - p53 expression as a prognostic indicator of 5-year survival in endometrial cancer. AB - BACKGROUND: One of the most common genetic alterations to occur in human cancers is an alteration of the p53 tumor suppressor gene. The purpose of this article was to build upon the authors' previous work with p53 and determine whether p53 was a prognostic indicator of 5-year survival. METHODS: One hundred thirty-seven consecutively surgically treated patients with endometrial cancer had their p53 expression studied by immunoperoxidase staining and quantified by image analysis. All patients were evaluable for 5-year survival. RESULTS: One hundred three patients had endometrioid adenocarcinoma; 6, adenosquamous carcinoma; 14, papillary serous carcinoma; 10, clear cell carcinoma; and 4, undifferentiated carcinoma. p53 expression ranged from 0.0 to 58.2% positive nuclear area with a mean of 11.5% (median 2.6%) for the cohort. For the patients with endometrioid carcinoma, the mean p53 expression was 7.1% while for the nonendometrioid tumors it was 24.6% (P<0.001). Fifty-nine of the 103 endometrioid tumors (57.3%) stained positive for p53 while 32 of the 34 nonendometrioid (94.1%) tumors stained positive (P<0.001). Increasing histologic grade correlated with an increasing p53 expression (P = 0.003). The percentage of tumors expressing p53 was found to be higher in FIGO stage II, III, and IV than in FIGO stage I cancer (P = 0.003). However, mean p53 expression did not differ between early (stage I) and advanced (stage II, III, and IV) cancers (P = 0.088). Utilizing 5-year survival as the endpoint for multivariate analysis, FIGO stage (P = 0.0028) and p53 expression (P<0.001) were the only independent prognostic indicators found. CONCLUSION: p53 expression is more commonly found in nonendometrioid than in endometrioid adenocarcinoma of the endometrium. It, along with FIGO stage, is an independent prognostic indicator of 5-year survival. PMID- 10479514 TI - Analysis of failed and complicated laparoscopy on a gynecologic oncology service. AB - OBJECTIVE: To assess the incidence of and factors that predict failed or complicated operative laparoscopy on a gynecologic oncology service. METHODS: Two hundred four consecutive operative laparoscopies were reviewed. Procedures converted to laparotomy or associated with major operative or postoperative complications were compared with uncomplicated laparoscopies with respect to patient characteristics, details of operative procedure, and length of hospitalization. The influence of patient characteristics, operative findings, and specific procedures on the risk of failed or complicated laparoscopies was estimated. RESULTS: Twenty-five (12. 3%) procedures were either converted to laparotomy or associated with major operative or postoperative complications. Women with failed or complicated laparoscopies had significantly more previous laparotomies and adhesions, greater blood loss, and longer hospital stay than those with uncomplicated laparoscopies (60.0% vs 35.7%, P = 0.03, 68.0% vs 37.4%, P<0.001, 275 ml vs. 132 ml, P = 0.03; and 5. 9 days vs 0.98 days, P< 0.001, respectively). Age, body mass index, parity, menopausal status, preoperative CA 125, appearance of adnexal masses, and complexity of the procedure had no significant influence on failed or complicated laparoscopies. In univariate analysis, history of laparotomy and presence of adhesions and in multivariate analysis only presence of adhesions were predictive of failed or complicated laparoscopies (P = 0.03, <0.001, and 0.006, respectively). CONCLUSIONS: The incidence of failed and complicated laparoscopy is low on a gynecologic oncology service. Presence of adhesions is the only significant independent risk factor predictive of failed or complicated laparoscopy. PMID- 10479515 TI - A patient with brain metastasis from ovarian cancer who showed complete remission after multidisciplinary treatment. AB - We describe a case with brain metastasis from ovarian cancer who showed complete remission after multidisciplinary treatment. The case was diagnosed as epithelial ovarian cancer, FIGO stage IIIc. She underwent cisplatin-based chemotherapy after optimal cytoreductive surgery and achieved clinical complete remission. Thirty two months after surgery for ovarian cancer, a solitary metastasis occurred in the left frontal lobe of the brain. No recurrent lesions were observed outside the brain. The metastatic tumor was resected. Five days after operation, adjuvant chemotherapy comprising carboplatin and cisplatin was initiated (a total of three courses at 4-week intervals), and whole brain irradiation at 55 Gy was added. After these treatment methods, complete remission of the brain metastasis has been observed for 57 months with good quality of life. PMID- 10479516 TI - Malignant Brenner tumor mimicking a primary squamous cell carcinoma of the cervix. AB - An 86-year-old female presented with a necrotic cervical mass that was biopsy proven squamous cell carcinoma. She had an elevated CA-125 and a pelvic mass. At surgery, this mass was found to be of adnexal origin and contiguous with the cervix. Histology showed a malignant Brenner tumor with abundant squamous differentiation eroding the cervix and simulating a primary cervical malignancy. We describe this case and review the literature on metastatic tumors to the cervix. PMID- 10479517 TI - Metastasizing signet ring cell carcinoma of the stomach-mimicking bilateral inflammatory breast cancer. AB - We describe a case of unusual metastases of a gastric carcinoma to the female breast, there likely to be an inflammatory breast cancer. A 46-year-old woman was admitted to our institution with bilateral breast tumors, not typical for a breast cancer as tumor growth was synchronically bilateral within a very short period of only 2 months. The woman underwent a palliative gastrectomy 3 months before for a poorly differentiated adenocarcinoma with signet ring appearance presenting as linitis plastica. At the time of the first operation the mammary glands were not suspicious. Breast biopsies assured metastases of the gastric cancer. In addition to this case report, a short overview of the literature concerning the very few cases of metastases of gastric cancer to the breast is given. PMID- 10479518 TI - Unusual recurrence of ovarian carcinoma 9 years after initial diagnosis. AB - Ovarian carcinomas typically metastasize to multiple sites via exfoliation, lymphatic spread, or direct invasion. Gastrointestinal tract involvement is usually the result of exfoliation with direct invasion of tumor within the mesentery or through serosal surfaces. We present a case of late recurrence of ovarian carcinoma isolated to the sigmoid mucosa, heralded only by brief left lower quadrant pain with hematochezia in a patient otherwise disease free for 9 years. This unusual presentation illustrates the therapeutic dilemma faced by clinicians when a tumor is of uncertain origin and underscores the need for continued follow-up and close scrutiny of new symptoms in patients with stage I disease and for those who enjoy prolonged disease-free intervals. PMID- 10479519 TI - Pleomorphic rhabdomyosarcoma of the uterus in a postmenopausal woman with elevated serum CA125. AB - BACKGROUND: Pure rhabdomyosarcoma of the uterus is an exceedingly rare tumor. While there are no commonly associated tumor markers, a previously reported case confirmed CA125 production by a pure uterine rhabdomyosarcoma. CASE: We describe a postmenopausal woman who presented with vaginal bleeding and was diagnosed with pleomorphic rhabdomyosarcoma of the uterus. Of particular interest was an elevated serum CA125 level at presentation. Immunohistochemical staining with CA125 antibody, however, showed no activity within the tumor. Despite aggressive therapy, the patient experienced distant nodal metastases and succumbed to the disease within 25 months of diagnosis. CONCLUSION: Our patient illustrates the typical presentation and aggressive behavior of this rare neoplasm. While serum CA125 may be a useful tumor marker in selected patients, elevated levels do not directly reflect tumor burden when associated with negative immunohistochemical staining. PMID- 10479520 TI - Chronic abdominal pain secondary to omental J-flap: report of two patients. AB - Two patients with a history of stage IB cervical cancer who had undergone prior radical abdominal hysterectomy, omental J-flap placement, and postoperative whole pelvic radiation therapy required subsequent exploratory laparotomy with resection of omental J-flap for treatment of chronic abdominal pain 5 and 2 years, respectively, following J-flap placement. Issues relating to this unusual possible long-term complication of omentoplasty are discussed. PMID- 10479521 TI - Florid endocervical glandular hyperplasia with intestinal and pyloric gland metaplasia: worrisome benign mimic of "adenoma malignum". AB - We describe three cases of florid endocervical glandular hyperplasia with intestinal and pyloric gland metaplasia, which can be a benign mimic of adenoma malignum. In two cases, adenoma malignum was seriously considered preoperatively because of watery vaginal discharge and the results of imaging studies. The three cases shared common histopathological features, i.e., (i) proliferating endocervical glands surrounded by clusters of smaller glands, resembling the pyloric glands of the stomach; (ii) occasional intestinal metaplasia; (iii) bland nuclear features; and (iv) predominantly PAS-positive neutral mucin in the glandular epithelium. In two cases, glands were densely and irregularly arranged in some areas. Immunohistochemistry disclosed that the intracytoplasmic mucin of the metaplastic epithelium was positive for M-GGMC-1 (HIK1083), which reacts with mucin of pyloric glands. Monoclonal CEA was negative in all cases. This pseudoneoplastic benign condition should be recognized by both gynecologists and pathologists, although it might be difficult to establish a definite diagnosis preoperatively even with deep cone biopsy. PMID- 10479522 TI - Trisomy 21 associated with ovarian dysgerminoma. AB - A 13-year-old G(0)P(0) white female with trisomy 21 presented with a complex pelvic mass. She underwent resection of the mass and complete staging for what was found to be a stage IIIC completely resected dysgerminoma. She was treated with three cycles of bleomycin, etoposide, and cisplatin chemotherapy and remains free of disease 1 year later. This association is presented as a rare case that may illustrate the relative increase in germ cell neoplasms in female patients with Down's syndrome. While the association of seminoma with Down's syndrome has been documented in a number of cases in males, the female counterpart of this tumor, dysgerminoma, in trisomy 21 has been reported quite infrequently. The potential for germ cell tumors in both male and female trisomy 21 is therefore illustrated. PMID- 10479523 TI - In vitro fertilization following conservative management of stage 3 serous borderline tumor of the ovary. AB - Among patients with advanced stage serous borderline tumors of the ovary, those with micropapillary architecture or invasive implants have the greatest risk of malignant transformation. In the absence of these patterns, consideration can be given to preservation of reproductive function. A 28-year-old, nulliparous patient presented with symptoms mimicking advanced ovarian cancer. Histology showed a serous borderline tumor with a hierarchical branching pattern. Surgery was able to remove all visible disease but still preserve the uterus and a portion of one ovary. She subsequently underwent in vitro fertilization and delivered a full-term infant. PMID- 10479524 TI - Comparison of molecular changes in cervical intraepithelial neoplasia in HIV positive and HIV-indeterminate subjects. AB - OBJECTIVE: HIV infection is associated with an increased incidence of cervical malignancy and its precursor lesions (CIN, cervical intraepithelial neoplasia) compared with the general population. We studied the molecular abnormalities in the development of HIV-associated CIN and compared them with those present in CINs arising in HIV-indeterminate subjects ("sporadic CIN"). METHODS: We investigated the presence of human papilloma virus (HPV) sequences, loss of heterozygosity (LOH), and microsatellite alterations (MAs) at five 3p chromosomal regions using 17 polymorphic markers in precisely microdissected archival tissues from 16 HIV-positive CINs and compared them with those present in 39 sporadic CINs. RESULTS: HPV sequences were detected in 36 of 55 (66%) CIN lesions, and high-risk oncogenic strains (HPV 16 and 18) accounted for 15 of them. No differences in the HPV frequencies were found between HIV-associated and sporadic CINs. Allelic losses at one or more chromosome 3p regions were frequently detected in CIN lesions (49%). The overall frequency of 3p LOH and the frequencies at all individual regions were similar in HIV-associated and sporadic CINs. The frequency of MA present in the HIV-associated CIN cases (0.093) was sixfold greater than in sporadic CINs (0.014; P = 0.0001). At least 1 MA was present in 11 (69%) of 16 HIV-associated vs. 5 of 39 (13%) sporadic CIN (P = 0.0006). Molecular changes were independent of the presence of HPV sequences. CONCLUSION: Chromosome 3p deletions are frequently detected in the precursor lesions of cervical carcinoma (CIN) and there are no differences in the 3p LOH frequencies between HIV-associated and sporadic CIN lesions. Microsatellite alterations, which reflect widespread genomic instability, occur at greatly increased frequency in HIV-associated CIN. Although the mechanism underlying the development of increased MAs is unknown, it may play a crucial role in the development of many HIV-associated neoplasias. PMID- 10479525 TI - The performance of colposcopy for women with atypical and low-grade cervical cytologic abnormalities. PMID- 10479526 TI - Current issues in cancer vaccine development. PMID- 10479527 TI - CD4(+) T cell clones producing both interferon-gamma and interleukin-10 predominate in bronchoalveolar lavages of active pulmonary tuberculosis patients. AB - The pattern of cytokine production in T cell clones derived from bronchoalveolar lavages (BAL) of active pulmonary tuberculosis (TB) patients was analyzed in clones obtained by limiting dilution procedures which expand with high efficiency either total T lymphocytes, independently of their antigen-recognition specificity, or Mycobacterium tuberculosis-specific T cells. BAL-derived clones, representative of CD4(+) cells from five patients with active TB, produced significantly higher amounts of IFN-gamma than BAL-derived CD4(+) clones from three inactive TB donors or four controls (with unrelated, noninfectious pathology). Average IL-4 and IL-10 production did not differ significantly in the three groups. Although these data suggest a predominant Th1 response to M. tuberculosis infection in the lungs, the majority of BAL-derived CD4(+) clones produced both IFN-gamma and IL-10 and the percentage of clones with this pattern of cytokine production was significantly higher in clones derived from BAL of active TB patients than from controls. Only rare clones derived from peripheral blood (PB)-derived CD45RO(+) CD4(+) T cells of both patients (nine cases) and controls (four cases) produced both IFN-gamma and IL-10; instead, the IL-10 producing clones derived from PB T cells most often also produced IL-4, displaying a typical Th2 phenotype. Higher average amounts of IFN-gamma and IL-10 were produced by BAL-derived CD8(+) clones of four active TB patients than of four controls, although the frequency of CD8(+) clones producing both IFN-gamma and IL-10 was lower than that of CD4(+) clones. The M. tuberculosis-specific BAL derived T cell clones from three active TB patients were almost exclusively CD4(+) and produced consistently high levels of IFN-gamma often in association with IL-10, but very rarely with IL-4. Unlike the BAL-derived clones, the M. tuberculosis-specific clones derived from PB CD45RO(+) CD4(+) T cells of three different active TB patients and two healthy donors showed large individual variability in cytokine production as well as in the proportion of CD4(+), CD8(+), or TCR gamma/delta(+) clones. These results indicate the predominance of CD4(+) T cells producing both the proinflammatory cytokine IFN-gamma and the anti inflammatory cytokine IL-10 in BAL of patients with active TB. PMID- 10479528 TI - Immune response enhancement by in vivo administration of B7.2Ig, a soluble costimulatory protein. AB - The identification of both class I- and class II-restricted tumor-associated peptides recognized by T cells has led to the test of these peptides as immunogens in experimental immunotherapy for cancer patients. However, optimal T cell activation requires signaling both through the T cell receptor for antigen and through costimulatory pathways. B7.1 and B7.2 are powerful costimulatory molecules expressed on the surface of antigen-presenting cells. Using a mouse model, we have sought to optimize costimulatory signals during antipeptide responses by administering a soluble form of B7.2 at the time of peptide immunization. Administration of B7. 2Ig fusion protein significantly enhanced T helper cell and CTL responses. These findings suggest that soluble forms of human B7.2 protein may provide a straightforward and practical method of supplying optimal costimulation during clinical immunotherapy. PMID- 10479529 TI - Cytokeratin 1 and gC1qR mediate high molecular weight kininogen binding to endothelial cells. AB - High molecular weight kininogen (HK) attaches to endothelial cells by separate sites on the heavy and light chains and requires 15-50 microM zinc. Previously identified binding proteins include gC1qR, cytokeratin 1, and the urokinase plasminogen activator receptor; however, their relative contributions to binding are not yet clarified. We have prepared affinity columns to which were coupled either cleaved HK or peptide LDCNAEVYVVPWEKKIYPTVNCQPLGM derived from heavy-chain domain 3. Endothelial cell membranes were solubilized and chromatographed in the presence or absence of zinc ion, the bound proteins were eluted, and active fractions were identified by dot blot using biotinylated HK, SDS/PAGE, and Western blot analysis. The peptide containing column eluate revealed but one band at 68 kDa if zinc ion was present which was identified as cytokeratin 1 by amino acid sequencing of an internal peptide. The HK affinity column revealed bands at 68 kDa (cytokeratin 1), 33 kDa (gC1qR), and 66 kDa (unidentified). HK or domain 3 derived peptide bound to the 68 kDa band; prekallikrein and Factor XII did not. HK or Factor XII bound to the 33-kDa band if zinc was present while no binding to the 66 kDa band was observed. Antibody to cytokeratin 1 inhibited HK binding to endothelial cells by 30%, antibody to gC1qR inhibited HK binding to endothelial cells by 72%, and a mixture of both inhibited binding by 86%. Our data suggest HK binding by interaction of the heavy-chain domain 3 with cytokeratin 1 and the light chain with gC1qR. PMID- 10479530 TI - Molecular definition and characterization of recombinant bovine CB8 and CB10: immunogenicity and arthritogenicity. AB - Theoretically, the ability to produce recombinant type II collagen (CII) peptide fragments in a prokaryotic expression system would be extremely useful for preparing adequate amounts of CII peptides suitable for therapeutic uses. Bacteria do not contain the enzymes involved in the extensive posttranslational modifications that occur during the biosynthesis of CII, such as the hydroxylation of prolyl and lysyl residues and glycosylation of hydroxylysyl residues. As these posttranslational modifications may play a role in the immune and arthritogenic response to CII, it was unclear whether collagen expressed in Escherichia coli would be immunologically comparable to tissue-derived CII. Therefore, we prepared recombinant proteins for CB8 and CB10 by cloning CB8 (CII 403-551) and CB10 (CII 552-897) genes from bovine chondrocytes by RT-PCR technique and expressing them in an E. coli expression system. Characterization of these recombinant proteins revealed that both rCB8 and rCB10 stimulated T cell proliferation in a T cell determinant-specific manner. The T cells from mice immunized with rCB8 respond specifically to a synthetic peptide, CII 445-453, the CB8 T cell determinant. Conversely, rCB10-primed T cells respond strongly to CII 610-618, the CB10 T cell determinant. Recombinant CB8-induced autoantibodies that bound to mouse CB8 as effectively and in the same topographic distribution as tissue-derived CB8. Finally, when rCB8 and rCB10 proteins were used to immunize B10.RIII (H-2(r)) mice, rCB8 induced arthritis in 33% of the mice, very similar to the incidence induced by tissue-derived CB8 peptide. As was found to be the case with tissue-derived CB10, rCB10 was completely ineffective in inducing arthritis. Pathological changes of arthritic joints in the mice immunized with rCB8 were similar to those observed in mice immunized with tissue-derived CB8. Thus, these recombinant CII peptides expressed in E. coli can induce an effective immunologic response and suggest that functionally useful CII peptides can be generated by the prokaryotic expression system. PMID- 10479531 TI - Salivary gland cytokine expression in NOD and normal BALB/c mice. AB - The autoimmune diabetes-prone nonobese diabetic (NOD) mouse develops a chronic lymphocytic infiltration of endocrine and exocrine glands. The objectives of this study were to characterize the salivary immune infiltration and cytokine expression of NOD mice and compare these findings to those of normal BALB/c mice. A decline in salivary flow rates in NOD mice began between 8 and 12 weeks of age. At this same time lymphocytic foci are detectable in the salivary glands. Lymphocytic infiltration in the salivary glands of NOD mice increased with age and simultaneously salivary function declined. No lymphocytic infiltration was seen in BALB/c salivary tissues. Messenger RNA expression of several inflammatory cytokines, including interleukin-1beta (IL-1beta), IL-2, IL-10, interferon-gamma, and tumor necrosis factor-alpha was detected in the submandibular glands of both NOD and BALB/c mice by the reverse transcription polymerase chain reaction. IL-4 synthesis was also present in some tissues. Immunohistochemical analysis demonstrated the intense expression of inflammatory cytokines within lymphocytic infiltrates and epithelial cells of all NOD mice. Minimal expression of the same cytokines was detected only occasionally in BALB/c tissues stained in parallel. These results demonstrate cytokine expression in the salivary glands of normal mice and suggest that the overexpression of these inflammatory cytokines is likely involved in the development and progression of the organ-localized autoimmunity in the salivary glands of NOD mice. PMID- 10479533 TI - Molecular mapping of the major epitopes of BP180 recognized by herpes gestationis autoantibodies. AB - Herpes gestationis (HG) is an autoantibody-mediated subepidermal bullous dermatosis associated with pregnancy. The primary target of HG autoantibodies is BP180, a 180-kDa hemidesmosomal glycoprotein. We previously showed that autoantibodies and autoimmune T lymphocytes from HG patients recognize the MCW-1 antigenic site (AA 507-520), which is located in the membrane-proximal noncollagenous domain (NC16A) of BP180. Here, we analyzed the sera of 37 HG patients to further define the sites on BP180 that are targeted by autoantibodies. All of the HG sera, but none of the control sera, were immunoreactive with sec180e, a 120-kDa recombinant protein encompassing the entire BP180 extracellular domain. HG sera depleted of reactivity to NC16A no longer reacted with sec180e, indicating that the major HG-associated epitopes on BP180 are restricted to the NC16A domain. The vast majority of the HG sera (34 of 37) reacted with a 7 amino acid peptide corresponding to the N-terminal half of MCW-1 (MCW-1A). Eleven HG sera (including the 3 that failed to react with MCW-1A) recognized one or more of three antigenic sites located within a 15 amino acid stretch immediately downstream of MCW-1A. In summary, we have identified four major HG-associated epitopes clustered within a 22 amino acid region of the BP180 ectodomain. These findings support the hypothesis that an autoimmune response to the BP180 NC16A domain is a crucial step in the pathogenesis of HG. PMID- 10479532 TI - Altered signaling lymphocytic activation molecule (SLAM) expression in HIV infection and redirection of HIV-specific responses via SLAM triggering. AB - Signaling lymphocytic activation molecule (SLAM) is a transmembrane lymphocytic receptor which gets rapidly upregulated following cell activation. SLAM engagement augments T cell expansion and interferon-gamma (IFN-gamma) production independently of CD28. SLAM signaling is regulated by the SLAM-associated protein. We evaluated the expression and function of SLAM on CD4(+) and CD8(+) lymphocytes in HIV-infected individuals with either recently acquired infection (Group A) or asymptomatic HIV infection (Group B) and in healthy controls (HC). Soluble antigen (HIV env peptides and tetanus toxoid)- and mitogen-stimulated proliferation and IFN-gamma and IL-10 production upon SLAM costimulation were also measured. Results showed that: (1) SLAM-expressing CD4(+) and CD8(+) lymphocytes diminish in group A patients compared to both group B patients and HC; (2) SLAM expression on CD4(+) lymphocytes is preferentially associated with the lack of CD7 on cell surface (CD4(+)CD7(-) produce IL-10 but not IFN-gamma); (3) SLAM engagement increases HIV env peptide-stimulated, but neither tetanus toxoid- nor PHA-stimulated proliferation of peripheral blood mononuclear cells (PBMC) in patients but not in HC; and (4) SLAM engagement augments IFN-gamma and reduces IL-10 production by env peptide-stimulated PBMC of HIV-infected individuals. These results demonstrate that early HIV infection results in an altered SLAM expression which correlates with a time-limited impairment of cell mediated immunity. Furthermore, they show that triggering via SLAM potentiates HIV-specific proliferative responses with simultaneous downregulation of IL-10 and redirection of the response to TH0/TH1. PMID- 10479534 TI - The development of AIDS-associated Burkitt's/small noncleaved cell lymphoma is preceded by elevated serum levels of interleukin 6. AB - B cell hyperactivation accompanies HIV infection and is believed to contribute to the increased incidence of B cell lymphoma in persons with AIDS. To examine B cell activation which precedes the development of AIDS-associated lymphoma, we measured levels of two B cell stimulatory molecules, soluble CD23 (sCD23) and interleukin 6 (IL6), in the serum of HIV-infected individuals prior to the diagnosis of lymphoma. Serum sCD23 was elevated in those subjects who developed lymphoma, compared to AIDS, HIV+, and HIV- controls (P = 0.001). Serum IL6 was significantly elevated in subjects who developed Burkitt's/small noncleaved cell lymphoma (BL/SNC, P = 0.01), but not in those subjects who developed large cell, immunoblastic, or central nervous system lymphomas, compared to CD4-matched AIDS controls who did not have lymphoma. These results suggest that lymphomagenesis of the BL/SNC subtype of AIDS lymphoma reflects B cell hyperactivation of a different nature from that which precedes other subtypes of AIDS-associated B cell lymphoma. PMID- 10479535 TI - Aberrant wound healing and TGF-beta production in the autoimmune-prone MRL/+ mouse. AB - Wound healing is a complex process that involves inflammation, apoptosis, growth, and tissue remodeling. The autoimmune-prone inbred mouse strain MRL/+ manifests accelerated and extensive healing to ear punch wounds, suggesting a link between immune defects and wound healing. Prior studies with lupus-prone mice have shown that hematopoietic cells of lupus-prone strains can transfer disease to otherwise non-autoimmune-prone recipients. In this study we performed reciprocal bone marrow transfers between MRL and the control strain B10.BR and found that radioresistant MRL/+ host cells, rather than hematopoietic cells, are required for the healing response. We have also made the novel observations that, compared to normal controls, MRL/+ hematopoietic cells overproduce TGF-beta1 and manifest impaired inflammatory responses to lipopolysaccharide challenge. These features suggest that the aberrant wound healing phenotype of MRL mice is independent of their propensity to develop autoimmunity. PMID- 10479536 TI - The biphasic mystery: why a biphasic shock is more effective than a monophasic shock for defibrillation. AB - We demonstrate that a biphasic shock is more effective than a monophasic shock at eliminating reentrant electrical activity in an ionic model of cardiac ventricular electrical activity. This effectiveness results from early hyperpolarization that enhances the recovery of sodium inactivation, thereby enabling earlier activation of recovering cells. The effect can be seen easily in a model of a single cell and also in a cable model with a ring of excitable cells. Finally, we demonstrate the phenomenon in a two-dimensional model of cardiac tissue. PMID- 10479537 TI - Epigenetic inheritance, genetic assimilation and speciation. AB - Epigenetic inheritance systems enable the environmentally induced phenotypes to be transmitted between generations. Jablonka and Lamb (1991, 1995) proposed that these systems have a substantial role during speciation. They argued that divergence of isolated populations may be first triggered by the accumulation of (heritable) phenotypic differences that are later followed and strengthened by genetic changes. The plausibility of this idea is examined in this paper. At first, we discuss the "exploratory" behaviour of an epigenetic inheritance system on a one peak adaptive landscape. If a quantitative trait is far from the optimum, then it is advantageous to induce heritable phenotypic variation. Conversely, if the genotypes get closer to the peak, it is more favorable to canalize the phenotypic expression of the character. This process would lead to genetic assimilation. Next we show that the divergence of heritable epigenetic marks acts to reduce or to eliminate the genetic barrier between two adaptive peaks. Therefore, an epigenetic inheritance system can increase the probability of transition from one adaptive state to another. Peak shift might be initiated by (i) slight changes in the inducing environment or by (ii) genetic drift of the genes controlling epigenetic variability. Remarkably, drift-induced transition is facilitated even if phenotypic variation is not heritable. A corollary of our thesis is that evolution can proceed through suboptimal phenotypic states, without passing through a deep adaptive valley of the genotype. We also consider the consequences of this finding on the dynamics and mode of reproductive isolation. PMID- 10479538 TI - Experimental realization of a signal transduction algorithm. AB - A theory of signal transduction specifies that in biological systems, any instantaneous input is appreciated by its departure from the mean past activity that occurred over a certain time window called the sample period; it also specifies that any input generates two so-called first- and second-order effects that are opposite in sign. We here report the detailed experimental realization of the algorithm that formalizes this signal transduction process; numerical simulations adequately predicted various intricate features of the (first order) analgesia and the paradoxical (second order) hyperalgesia which morphine produces. The data also offer a first estimation of the physiological sample period that may govern the classical rat tail flick response. The signal transduction process appears to operate ubiquitously and provides an unprecedented account of the paradoxical effects that have been observed with different signalling systems. This process may also operate with such phenomena as refractoriness, homeostasis, adaptation, sensitization, dependence, tolerance or resistance and neuronal plasticity. PMID- 10479539 TI - Neutral evolution of model proteins: diffusion in sequence space and overdispersion. AB - We stimulate the evolution of model protein sequences subject to mutations. A mutation is considered neutral if it conserves (1) the structure of the ground state, (2) its thermodynamic stability and (3) its kinetic accessibility. All other mutations are considered lethal and are rejected. We adopt a lattice model, amenable to a reliable solution of the protein folding problem. We prove the existence of extended neutral networks in sequence space-sequences can evolve until their similarity with the starting point is almost the same as for random sequences. Furthermore, we find that the rate of neutral mutations has a broad distribution in sequence space. Due to this fact, the substitution process is overdispersed (the ratio between variance and mean is larger than 1). This result is in contrast with the simplest model of neutral evolution, which assumes a Poisson process for substitutions, and in qualitative agreement with the biological data. PMID- 10479540 TI - A baseline model for the apparent competition between many host strains: the evolution of host resistance to microparasites. AB - The purpose of this article is to establish and analyse a baseline model for the apparent competition between many host strains attempting to avoid a uniform microparasitic population. The model is formulated and analysed using invasion criteria in the main text. The results are verified by more formal methods in the appendix. Cases in which the microparasite can invade are distinguished geometrically from those in which it cannot using threshold and strain composition conditions. A major result obtained when the pathogen persists is a competitive exclusion principle for host resistance. For non-lethal infections, the winning strain is that which affords the pathogen maximum threshold density; for possibly lethal infections, a somewhat generalized version of this criterion is presented and discussed. The tension is highlighted between these results and the baseline behaviour of many pathogen strains and a uniform host population here the winning pathogen strain is that with minimum threshold density. PMID- 10479541 TI - Biological probability: cognitive processes of generating probabilities of events in biological systems. AB - This paper analyses relationships between probabilities of events happening in biological systems (or probabilistic disposition of systems) and cognitive properties of biological entities comprising such systems. Two kinds of cognitive properties are identified as relevant to the current problem: the ability to respond differently against different configurations of the environment (discriminability of cognition), and the ability to make an appropriate response to maintain a particular relation with the environment (selectivity of cognition). A basic framework bridging the two features of living systems, probabilistic disposition and the cognitive properties, is presented towards a general theory explaining the process generating probabilities of biological events. In this framework, a deterministic model of a system of entities is developed, in which objects are described as subjects that cognize events (i.e. entities as cognizers). Cognition is used in a wider sense, including not only biotic but also abiotic, and cognizers are conceptually distinguished from the meta-observer who describes the system externally. Based on this perspective, this paper seeks to explicate how events can occur in an uncertain, probabilistic manner, if observed from a cognizer viewpoint, even under a deterministic system. Each cognizer is identified with both the set of states that are actually taken, and its motion function which maps its state uniquely to a successor state depending on the current states of itself and of the rest of cognizers constituting the system. The model analysis reveals that the cognitive properties, discriminability and selectivity, of a cognizer can contribute to determining the probability of an event encountered by the cognizer itself-in particular, discrimination reducing the uncertainty in events occurrence for the cognizer. Biological implication of this result is discussed focusing on the concept of the probability of survival and reproduction. PMID- 10479542 TI - Good parent and good genes models of handicap evolution. AB - We previously studied a good genes handicap model in which male quality was heritable and improved offspring viability. We extend our analysis to species in which males provide direct benefits (e.g. parental care, better resources, the absence of contagious diseases). Male quality now affects female fitness by increasing female reproductive success. For this good parent handicap to work, the male signal must have condition-dependent expression. The equilibrium strength of female preference is controlled by the product of signal transmission efficiency, phenotypic variance of male quality and the effectiveness of male quality in improving female reproductive success. The equilibrium resulting from the good parent handicap has exactly the same form as with the good genes handicap. This allows us to compare the relative importance of these two forces in the evolution of female preferences. The handicap models (both good genes and good parent) also show cyclic evolution, as happens with the pure Fisherian model. However, we predict that the handicap process is often strong enough to lead to a stable equilibrium. This leads to the conclusion that cyclic evolution is less likely to occur for handicap than Fisherian traits. PMID- 10479543 TI - Persistence of tick-borne virus in the presence of multiple host species: tick reservoirs and parasite mediated competition. AB - Tick-borne viruses in tropical and temperate parts of the world have a significant impact on human, livestock and wildlife hosts both directly, through mortality/morbidity, and economically. Since the ticks have multiple life stages and can utilize a large range of host species our understanding of the dynamics of these infections is often not clear. In this paper we consider the impact of a population which is a tick host but non-viraemic on one which is both a tick host and viraemic. We present two simple deterministic models and use joint threshold density curves to illustrate the basic reproductive ratios of both the ticks and the virus. We find that the non-viraemic hosts can have considerable impact on the viraemic host. Either they amplify the tick population and cause the virus to persist, or they dilute the infection and cause it to die out. A general model framework is presented here but a special case of this model describes the red grouse-hare-Louping-ill system. PMID- 10479544 TI - Cross-correlated quadrupolar spin relaxation and carbon-13 lineshapes in the (13)CD(2) spin grouping. AB - Lineshape calculations are reported for a spin system consisting of a spin-12 nucleus scalar-coupled to two magnetically equivalent spin-1 nuclei, e.g., a CD(2) group. It is found that, because of the differential line broadening, the peak height ratio of the five spin-12 nucleus transitions is expected to deviate from the integrated intensity ratio of 1:2:3:2:1. The deviation is dependent on the extent of cross-correlation between the two quadrupolar interactions. The theoretical predictions for peak height ratios and for bandshapes are tested for the carbon-13 spectrum of perdeuterated ethylene glycol, and iterative fitting is used to obtain an estimate of the extent of the correlation between the quadrupolar interactions for the two deuterons. PMID- 10479545 TI - Spectroscopy from proton spin magnetization evolution in their rotating frame: A study of small tunneling splitting. AB - The time evolution of proton Zeeman magnetization in the rotating frame at exact resonance, omega = omega(0), is evaluated for an isolated tunneling methyl group CH(3). The Fourier transform of this evolution in time is calculated and both its real and imaginary components are presented. It is shown that the real component does not depend significantly on the strength of the preparation pulse when the tunneling splitting of the methyl rotator ground state is less than 100 kHz. It is also found that the imaginary component of the transform is inversely proportional to the strength of the preparation RF pulse. This is a consequence of the partial dephasing of proton spins during the preparation pulse. The results of the calculation compare well with the experimental spectra of CH(3)CD(2)I. PMID- 10479546 TI - Error-tolerant RF litz coils for NMR/MRI. AB - A new class of NMR RF volume coils is being developed that permits improved tuning range, B(1) homogeneity, tuning stability, and sensitivity compared to birdcages over a wide range of practical conditions, especially for microscopy and wraparound flexible applications. They are denoted litz coils, as their flux transparency and current distribution is obtained from woven foil patterns with insulated crossovers. Contrary to the design criteria of phased arrays, the parallel routes in litz coils use high coupling coefficients to achieve optimal current distribution, which is highly independent of tuning, balancing, and matching adjustments and is compatible with multiple capacitive segmentation. Magnetic filling factors, loaded Q, and inhomogeneity measurements and calculations are presented for a variety of litz coils with frequency-diameter products from 7 to 20 MHz-m and are compared to similar birdcages. PMID- 10479547 TI - Measurement of (1)J(NC') and (2)J(H(N))(C') couplings from spin-state-selective two-dimensional correlation spectrum. AB - A method for the measurement of (1)J(NC') and (2)J(H(N))(C') coupling constants from a simplified two-dimensional [(15)N, (1)H] correlation spectrum is presented. The multiplet components of the (1)J(NC') doublet in the indirect dimension and (2)J(H(N))(C') in the direct dimension are separated into two subspectra by spin-state-selective filters. Thus each subspectrum contains no more peaks than the conventional [(15)N, (1)H]-HSQC spectrum. Furthermore, the method for the measurement of (1)J(NC') and (2)J(H(N))(C') is designed to exploit destructive relaxation interference (TROSY). The results are verified against the measurements of (1)J(NC') from spin-state-selective [(13)C', (1)H] correlation spectra recorded with additional sequence described here. PMID- 10479548 TI - High-resolution strain analysis of the human heart with fast-DENSE. AB - Single breath-hold displacement data from the human heart were acquired with fast DENSE (fast displacement encoding with stimulated echoes) during systolic contraction at 2.5 x 2.5 mm in-plane resolution. Encoding strengths of 0.86-1.60 mm/pi were utilized in order to extend the dynamic range of the phase measurements and minimize effects of physiologic and instrument noise. The noise level in strain measurements for both contraction and dilation corresponded to a strain value of 2.8%. In the human heart, strain analysis has sufficient resolution to reveal transmural variation across the left ventricular wall. Data processing required minimal user intervention and provided a rapid quantitative feedback. The intrinsic temporal integration of fast-DENSE achieves high accuracy at the expense of temporal resolution. PMID- 10479549 TI - An optimized pulse sequence for isotropically weighted diffusion imaging. AB - Single-shot echo-planar imaging is becoming the most widely used technique for magnetic resonance diffusion imaging, since it enables measurement of diffusion coefficients in human brain without motion artifacts. However, its reliability is limited by geometrical distortions due to eddy currents. In this report, an isotropically weighted echo-planar pulse sequence, optimized to give the maximum signal-to-noise ratio in the computed trace image and designed to produce inherently low distortions, is presented. It is also shown how the residual translational distortion can be easily characterized and removed by postprocessing. A full characterization of the distortion artifact involves a few measurements on a phantom, in order to estimate the distortion as a function of slice orientation, which can then be used to correct any slice orientation. Results of applying the image translation correction to data collected from a patient are presented. PMID- 10479550 TI - Absolute EPR spin echo and noise intensities. AB - EPR signal and noise, calculated from first principles, are compared with measured values of signal and noise on an S-band (ca. 2.7 GHz) EPR spectrometer for which all relevant gains and losses have been measured. Agreement is within the uncertainty of the calculations and the measurements. The calculational model that provided the good agreement is used to suggest approaches to optimizing spectrometer design. PMID- 10479551 TI - Feedback optimization of pulse width in the SORC sequence. AB - A method for increasing the signal-to-noise ratio (SNR) of nuclear quadrupole resonance (NQR) measurements by automatically adjusting a pulse parameter in real time is presented. This approach is useful in situations where the optimal pulse parameters cannot be chosen beforehand due to lack of knowledge regarding the system. For example, NQR provides a means for detecting explosives by revealing the presence of (14)N. In this particular application, the distance between the search coil and the explosive, as well as the temperature of the explosive, is unknown. As a result, a fixed set of pulse parameters will not yield the largest SNR for all possible search applications. This paper describes a feedback algorithm that uses measurements of the NQR signal to automatically adjust the pulse width in the strong off-resonant comb sequence to maximize the SNR of the NQR measurement. Experimental results obtained using a sample of sodium nitrite are presented. PMID- 10479552 TI - General analysis of (14)N (I = 1) electron spin echo envelope modulation. AB - The analysis methods described to date for (14)N electron spin echo envelope modulation (ESEEM) mostly deal with isotropic g- and (14)N hyperfine coupling tensors. However, many cases of rhombic tensors are encountered. In the present report we present general equations for analyzing orientation-selective ESEEM and illustrate their use. (i) We present general equations for the nuclear interactions in an electron spin system where the EPR signal arises from an isolated Kramers doublet, then give the nuclear (electron-nuclear double resonance) frequencies for I = 1 associated with such a system. (ii) These are incorporated into equations for single-crystal ESEEM amplitudes, which in turn are incorporated into general equations for the orientation-selective ESEEM that arises when the EPR envelope of a frozen-solution (powder) sample is determined by g anisotropy. (iii) This development is first used in the simplest limit of an isotropic g-tensor and leads to a more general picture of the response of the I = 1 modulation amplitude to variations in the nuclear hyperfine and quadrupole coupling constants, relative to the nuclear Zeeman interaction, than had been presented previously. We find that strong modulation occurs not only in the well known regime where the "exact/near cancellation" condition (A/2 approximately nu(N)) is satisfied, but also when the nuclear hyperfine interaction is much larger than the nuclear Zeeman interaction (A/nu(N) > 3) with A/K = 4 approximately 5. (iv) We then describe the orientation-selective (14)N ESEEM frequency-domain patterns (g vs frequency) in the presence of anisotropic (rhombic) hyperfine and electron Zeeman interactions for both coaxial and noncoaxial cases. We derive analytical solutions when the g-, hyperfine, and nuclear quadrupole tensors are coaxial. (v) The method is applied to the ESEEM of the nitrogenase MoFe protein (Av1) to determine the full hyperfine and nuclear quadrupole tensors of (14)N nuclei interacting with the S = 32 FeMo-cofactor (Fe(7)S(8)Mo: homocitrate). PMID- 10479553 TI - Spectral fitting of NMR spectra using an alternating optimization method with a priori knowledge. AB - As alternatives to the fast Fourier transform, advanced parametric methods based on the damped sinusoidal data model have been devised to better quantify the nuclear magnetic resonance (NMR) spectroscopy time-domain data. Previously, linear prediction (LP) fitting methods using Householder triangularization and singular value decomposition (SVD) techniques have been applied to the NMR spectroscopy data analysis. In this paper, we propose an alternating optimization method to quantify the time-domain NMR spectroscopy data. The proposed algorithm uses the a priori knowledge of the possible frequency intervals of the damped sinusoids to obtain more accurate parameter estimates when the NMR spectroscopy data are obtained under low signal-to-noise ratio conditions and the peaks are close together. None of the LP and SVD type of methods can use such approximate a priori knowledge. We have shown with measured NMR spectroscopy data that the proposed algorithm can be used to obtain accurate parameter estimates of frequencies, amplitudes, and damping ratios of the damped sinusoids and therefore the ultimate fit of the spectrum by using the a priori knowledge about the possible frequency intervals of the damped sinusoids. PMID- 10479555 TI - Experimental aspects of multidimensional solid-state NMR correlation spectroscopy. AB - The experimental parameters critical for the implementation of multidimensional solid-state NMR experiments that incorporate heteronuclear spin exchange at the magic angle are discussed. This family of experiments is exemplified by the three dimensional experiment that correlates the (1)H chemical shift, (1)H-(15)N dipolar coupling, and (15)N chemical shift frequencies. The broadening effects of the homonuclear (1)H-(1)H dipolar couplings are suppressed using flip-flop (phase and frequency-switched) Lee-Goldburg irradiations in both the (1)H chemical shift and the (1)H-(15)N dipolar coupling dimensions. The experiments are illustrated using the (1)H and (15)N chemical shift and dipolar couplings in a single crystal of (15)N-acetylleucine. PMID- 10479554 TI - Time-domain quantification of series of biomedical magnetic resonance spectroscopy signals. AB - Quantification of individual magnetic resonance spectroscopy (MRS) signals is possible in the time domain using interactive nonlinear least-squares fitting methods which provide maximum likelihood parameter estimates under certain assumptions or using fully automatic, but statistically suboptimal, black-box methods. In kinetic experiments time series of consecutive MRS spectra are measured in which information concerning the time evolution of some of the signal parameters is often present. The purpose of this paper is to show how AMARES, a representative example of the interactive methods, can be extended to the simultaneous processing of all spectra in the time series using the common information present in the spectra. We show that this approach yields statistically better results than processing the individual signals separately. PMID- 10479556 TI - On the generation of operator equivalents and the calculation of their matrix elements. AB - To find all components T((k))(+/-q) = N(k,q)J(q)(+/-) summation operator(k q)(m=0) (+/-1)(k-m)a(k, q; m)J(m)(z) (0 0), (2 --> 1), (0 <-- 1), (1 <-- 2) bands of the B(3)Pi(g)-W(3)Delta(u) system, not included in any previous analysis. Spectroscopic parameters for the v = 0, v = 1, v = 2 levels of the B(3)Pi(g) and the W(3)Delta(u) states, consistent with those previously reported but with improved accuracy, are obtained from the experimental wavenumbers by a nonlinear least-squares procedure. Copyright 1999 Academic Press. PMID- 10479582 TI - Line Positions and Strengths of N(2)O between 3515 and 7800 cm(-1). AB - The line positions and strengths of N(2)O were measured from absorption spectra obtained at high spectral resolution (0.011 cm(-1)) in the region between 3515 and 7800 cm(-1). Measurements include line frequencies and strengths of several N(2)O isotopes, a few of which are reported for the first time, to my knowledge. Also in this category are two pi-pi bands, six Delta-Delta bands, and five Sigma Sigma bands of (14)N(2)(16)O. Measurements also include eight (14)N(2)(16)O bands of which several transitions of each band were perturbed due to resonance interactions with transitions of a perturbing band. For most cases, the positions and strengths of the strongest interacting transitions of the perturbing bands were measured. Copyright 1999 Academic Press. PMID- 10479583 TI - Identification of New Hot Bands in the Blue and Green Band Systems of FeH. AB - A particularly rich region of the electronic spectrum of FeH from 525 to 545 nm was investigated using the techniques of dispersed and undispersed laser-induced fluorescence. Analysis has led to the discovery that several different electronic transitions are embedded in this region; the (0, 0) and (1, 1) bands of the e(6)Pi-a(6)Delta (green) system, the (0, 2) band of the g(6)Phi-X(4)Delta (intercombination) system, the (0, 1) band of the g(6)Phi-a(6)Delta (blue) system, and the (0, 0) band of the g(6)Phi-b(6)Pi system. Seventy-five lines were assigned in the (0, 1) band of the g(6)Phi-a(6)Delta transition. These, with the assignment of an additional 14 lines in the 583 nm region to the (0, 1) band of the e(6)Pi-a(6)Delta transition, led to the extension of the known term values to higher J values for the Omega = 9/2, 7/2, and 5/2 spin components of the v = 1 level of the a(6)Delta state and the novel characterization of the a(6)Delta(3/2) (v = 1) and g(6)Phi(5/2) (v = 0) components. A further 73 lines were assigned to the first four subbands of the (1, 1) band of the e(6)Pi-a(6)Delta transition and term values for the lowest four spin components of the v = 1 level of the e(6)Pi state were determined. This provides the first experimental measurement of a vibrational interval in one of the higher lying electronic states of FeH. The interval does not appear to vary strongly between the spin components (DeltaG(1/2) = 1717, 1713, 1710 cm(-1) for Omega = 7/2, 5/2, 3/2, respectively). Remarkably few of the hot-band transitions assigned in this work could be identified in the complex, high-temperature spectrum of FeH recorded by P. McCormack and S. O'Connor [Astron. Astrophys. Suppl. 26, 373-380 (1976)]. Copyright 1999 Academic Press. PMID- 10479584 TI - The 3A Band System in the Spectrum of the (13)C(16)O Molecule. AB - In the emission spectrum of the carbon monoxide (13)C(16)O isotopic molecule three bands comprising about 1820 lines of the 3A band system (c(3)Pi-a(3)Pi) were recorded and analyzed. The 0-0 and 0-1 bands of this system were photographed for the first time and the 0-2 band was rephotographed by using methods of conventional high-resolution spectroscopy. The result of the rotational band analysis includes expanding of the spectrum interpretation up to J = 25 as well as the identification of four previously unobserved branches P(13), R(13), P(31), and R(31). Because of strong perturbations in the c(3)Pi (v = 0) state, the calculation of the rovibronic structure constants was performed only for the lower a(3)Pi state. By using a calculation based on a nonlinear least-squares method, an effective Hamiltonian of Brown [J. M. Brown, E. A. Colbourn, J. K. G. Watson, and F. D. Wayne, J. Mol. Spectrosc. 74, 294-318 (1979)] and a separative procedure proposed by Curl-Dane-Watson [R. F. Curl and C. B. Dane, J. Mol. Spectrosc. 128, 406-412 (1988); J. K. G. Watson, J. Mol. Spectrosc. 138, 302-308 (1989)], it was possible to derive the rotational structure constants for the v = 0, 1, and 2 levels for the a(3)Pi state in the (13)C(16)O isotopic molecule. Term values for the c(3)Pi (v = 0) level and the equilibrium molecular constants for the a(3)Pi state also are reported. Copyright 1999 Academic Press. PMID- 10479585 TI - The Ground State of D(2)Se and HDSe. AB - Ground state rotational constants of D(M)(2)Se and HD(M)Se, M = 76, 77, 78, 80, and 82, have been determined up to octic centrifugal distortion terms from ground state combination differences. These were obtained from rotational analyses of the nu(2), nu(1), and nu(3) bands both of natural and (80)Se monoisotopic material recorded with a resolution of ca. 3 x 10(-3) cm(-1). While the full set of rotational parameters of the (80)Se species was determined with significance, some of the centrifugal distortion terms of the less abundant species were either constrained to those of the (80)Se species or extrapolated. Copyright 1999 Academic Press. PMID- 10479586 TI - The nu(2) Bands of the Deuterated Species D(2)Se and HDSe. AB - For the first time D(2)Se and HDSe as (80)Se monoisotopic and natural material were studied in the region of the nu(2) fundamental vibration by high-resolution (0.0033 cm(-1)) Fourier transform infrared spectroscopy. For D(2)Se which is an asymmetric rotor with C(2v) symmetry the nu(2) band is of B type while for HDSe (C(s) symmetry) it is a hybrid band, and both A- and B-type transitions were observed. Depending on the isotopic abundances, 300-1000 lines were assigned for each of the isotopic (M)Se species (M = 76, 77, 78, 80, and 82). The corresponding (010) experimental rotational levels, obtained by adding the observed line positions to the ground state levels derived from J.-M. Flaud, Ph. Arcas, O. N. Ulenikov, G. A. Onopenko, N. E. Tyabaeva, W. Jerzembeck, and H. Burger, (J. Mol. Spectrosc., in press) were fitted satisfactorily using a Watson type Hamiltonian in A reduction and I(r)-representation: the rms deviations are ranging from 1.5 to 4.3 x 10(-4) cm(-1) depending on the isotopic species. Hamiltonian constants up to high order (octic terms) were determined for each isotopic species. For the most abundant Se isotopic species, namely (80)Se, the band centers are nu(0) (D(2)(80)Se) = 741.67503 cm(-1) and nu(0) (HD(80)Se) = 900.43820 cm(-1). Copyright 1999 Academic Press. PMID- 10479587 TI - Development of the Dipole Moment and Polarizability Operators of Octahedral Molecules. AB - We present a development of the dipole moment and polarizability operators of octahedral molecules, using a tensorial formalism analogous to the one developed for tetrahedral molecules. These operators are involved in the calculation of the intensities of rovibrational transitions as well as in the calculation of the Stark effect. Expressions for the matrix elements are derived. Two simplified models for the study of the Stark effect in such molecules are also proposed and discussed. Copyright 1999 Academic Press. PMID- 10479588 TI - The Rotational Spectrum of Ar-SiH(4) and Ar-SiD(4). AB - Microwave spectra of Ar-(28)SiH(4), Ar-(29)SiH(4), Ar-(30)SiH(4), and Ar (28)SiD(4) were recorded using a pulsed molecular beam Fourier transform microwave spectrometer. The K = 0 and K = 1 components of the J = 3 <-- 2 through the J = 7 <-- 6 transitions were measured and assigned in the 9-24 GHz region. For the primary (28)Si isotopic species, Ar-(28)SiH(4) and Ar-(28)SiD(4), a K = 0, A symmetry, a K = 0, F symmetry, a doubly degenerate K = 1, E symmetry, and an l/K-doubled, K = 1, F symmetry rotational progression are observed at the approximately 1 K rotational temperature of the supersonic expansion. The rotational constants for the K = 0, A state for Ar-(28)SiH(4) and Ar-(28)SiD(4) are B = 1700.40624(9) MHz and 1630.687073(22) MHz and the centrifugal distortion constants are D(J) = 29.089(3) and 20.0198(8) kHz and H(J) = -1.91(3) and 0.851(8) Hz, respectively, where type A expanded uncertainties with a coverage factor, k = 3, are given here and elsewhere. The values of the rotational constants for the K = 0, A, and F states and for the K = 1, E state are in good agreement with the infrared-determined values for Ar-(28)SiH(4). The measured linear Stark effect of the E-state transitions was analyzed to give a dipole moment of 9.24(8) x 10(-32) C. m [0.0277(2) D]. Copyright 1999 Academic Press. PMID- 10479589 TI - The VUV Absorption Spectrum of the GeO Molecule. AB - The absorption spectrum of GeO, between 2000 and 300 A, was recorded using the facilities of the SUPERACO synchrotron radiation in Orsay, France. The first three ionization limits, corresponding to the X(2)Sigma(+), A(2)Pi, and B(2)Sigma(+) states of the GeO(+) ion, were confirmed at 11.2814, 11.42, and 15.048 eV, respectively. The fourth ionization limit corresponding to the C(2)Sigma(+) ionic state was measured for the first time at 31.47 eV, in reasonable agreement with theoretical calculations. Copyright 1999 Academic Press. PMID- 10479590 TI - Rotational Spectroscopic Investigation of the Weak Interaction between CO and N(2)O. AB - Pure rotational spectra of the CO-N(2)O complex are reported for the first time. Spectra of five isotopomers, i.e., (12)C(16)O-(14)N(14)NO, (13)C(16)O (14)N(14)NO, (13)C(18)O-(14)N(14)NO, (13)C(16)O-(15)N(14)NO, and (13)C(16)O (14)N(15)NO, were measured using a pulsed molecular beam Fourier transform microwave spectrometer. Rotational and quartic centrifugal distortion constants as well as the nuclear quadrupole coupling constants of the terminal and the central (14)N nuclei were determined. Comparisons of the nuclear quadrupole coupling constants with the corresponding values of the N(2)O monomer and with those of other N(2)O-containing complexes indicate significant electronic charge redistribution at the central nitrogen atom upon complex formation. Structural parameters based on the experimental rotational constants were derived. With the isotopic data available, the orientations of the CO and N(2)O subunits in the complex could be determined. Copyright 1999 Academic Press. PMID- 10479591 TI - The Reduced Effective Vibration-Rotational Hamiltonian for the v(t) = 2 Levels of C(3v) Symmetric-Top Molecules. AB - The theory of the reduction of the effective rotation-vibration Hamiltonian is applied to the v(t) = 2 states of C(3v) molecules, and the higher-order terms up to H(43) are included. The results explain clearly why it is possible to determine in this state all four spectroscopic constants eta(J), eta(K), q(12), and d(t) simultaneously without the indeterminacy problems that have been encountered in the v(t) = 1 states. Constraints suitable for most cases are proposed and alternative choices are discussed. A number of new relations among the parameters of the various reduced forms of the effective Hamiltonian is predicted. Copyright 1999 Academic Press. PMID- 10479592 TI - A Theoretical Investigation of the Renner Interactions and Magnetic Dipole Transitions in the A-&Xtilde; Electronic Band System of HO(2). AB - The A(2)A' --> &Xtilde;(2)A" electronic band system of HO(2) has been simulated in emission using an extended version of the program RENNER (P. Jensen, M. Brumm, W. P. Kraemer, and P. R. Bunker, J. Mol. Spectrosc. 171, 31-57 (1995)). The two electronic states involved in this transition have strongly bent equilibrium geometries but they correlate together to form a (2)Pi state at linearity. As a result the energy level pattern in the states is affected by electronic angular momentum effects (i.e., the Renner effect and spin-orbit coupling). To simulate the spectrum, we have calculated ab initio the potential energy surfaces, electric dipole moment surfaces, magnetic dipole moment surfaces, spin-orbit coupling parameter, and the electronic angular momentum matrix elements. Some of the forbidden DeltaK(a) = 0 transitions occurring in the spectrum are induced by the magnetic dipole transition moment, and the others are electric dipole transitions that gain intensity because of the Renner interaction, spin-orbit coupling, or because of rotation-vibration interaction. All of these effects are allowed for in our calculation. The electric dipole transition moment is very small (0.017 D at the ground state equilibrium geometry) and because of this the magnetic dipole transitions are quite visible; the strongest magnetic dipole transitions are calculated to be about 10 times weaker than the strongest electric dipole transitions. In this way previous experimental assignments (E. H. Fink and D. A. Ramsay, J. Mol. Spectrosc. 185, 304-324 (1997)) are confirmed theoretically. Copyright 1999 Academic Press. PMID- 10479593 TI - The Ground and First Torsional States of CD(3)CHO. AB - The rotational spectra for the ground and first excited torsional states v(t) = 0 and 1 in the frequency region of 8-254 GHz and the v(t) = 1 <-- 0 band high resolution far-infrared spectrum of 2,2,2-d(3)-acetaldehyde (CD(3)CHO) were measured. We fitted a data set consisting of 1016 v(t) = 1 <-- 0 far-infrared lines together with 195 microwave lines in v(t) = 0 and 79 microwave lines in v(t) = 1 to near-experimental accuracy, using a global model from the earlier literature. The final fit includes lines with J /=18 years, 12.9% (95% confidence interval [95% CI] = 10.1-15. 6) received a Pap test reminder within the past year, while 13.0% (95% CI = 9.3-16.7) of women aged >/=40 years received a mammography reminder. Women without health care coverage were unlikely to receive either type of reminder. Current compliance with screening recommendations was greater among those women who received a reminder letter for Pap tests (94.3 versus 78.1%, P < 0.0001) and for mammography (81.7 versus 59.4%, P < 0.001). In contrast to the infrequent use of cancer screening reminders, 54.2% (95% CI = 50. 1-58.3) and 72.7% (95% CI = 67.6-77.8) of women reported receiving a reminder letter from their dentist or veterinarian, respectively. CONCLUSIONS: Reminder letters for cancer screening services were rarely utilized in this study population. Receipt of a reminder letter was associated with greater compliance with current screening recommendations. PMID- 10479609 TI - Cigarette smoking among medical college students in Wuhan, People's Republic of China. AB - BACKGROUND: This study examined cigarette smoking among a sample of medical college students in Wuhan, People's Republic of China. Specific cultural factors associated with cigarette smoking in China were discussed and issues which need to be studied in the future are presented. METHODS: A self-administrated questionnaire was administered to a sample of 1,611 medical students in 1995. Smoking status was based on self-reported use of cigarettes within the preceding 180 days. Information about demographic characteristics, home residence, annual family income, and smoking status of family members was also obtained. RESULTS: A total of 1,540 (1,201 males and 339 females) completed and returned the questionnaires (response rate 95.6%). None of the 339 female students surveyed reported cigarette smoking. A total of 37.7% (95% confidence interval (CI) = 35.0 40.5%) of the male medical students were current smokers, and 5. 3% (95% CI = 4.0 6.6%) were former smokers. The mean age of beginning to smoke was 16.5 years (95% CI = 8.7-24.3 years). The major reasons for first smoking were stress (42.8%, 95% CI = 38.2-47. 4%), curiosity (34.4%, 95% CI = 29.6-38.4%), and loneliness (33.7%, 95% CI = 28.7-37.4%). Multivariate analyses suggested that age, college year, and having a family member who smoked were significantly associated with cigarette smoking. CONCLUSIONS: The results indicated a need to provide comprehensive tobacco use prevention programs among medical students. PMID- 10479610 TI - Bilateral prophylactic mastectomy decision making: A vignette study. AB - BACKGROUND: Little is known about the perception of bilateral prophylactic mastectomy (BPM), and whether perceptions are influenced by a family history of breast cancer. It is also unclear what factors may play a role in selecting BPM for follow-up care. METHODS: Samples of predominantly Caucasian, well-educated women with (n = 129) and without (n = 104) family histories of breast cancer were provided a vignette of a woman at increased risk. They selected one of two follow up options: (1) clinical breast examination, breast self exam, and annual mammography or (2) BPM. RESULTS: The samples did not differ on the decision to select BPM (29.5% vs 22.1%). The family history sample reported worry about breast cancer as a problem (34.4%) more often than women with no history (15.7%). Multivariate analysis found worry and estimated 10-year risk of the woman in the vignette as significant predictors of selecting BPM. CONCLUSIONS: Approximately 25% of our sample selected BPM as the preferred option. The majority supported the need to discuss BPM among women at increased risk. Finally, both factors associated with the selection of BPM (worry, risk assessment) are potentially amenable to psychosocial or educational approaches. There is a clear need for assessment of worry and risk perception prior to surgical decision making. PMID- 10479611 TI - Radon-smoking synergy: A population-based behavioral risk reduction approach. AB - BACKGROUND: Radon and cigarette smoking have a synergistic, multiplicative effect on lung cancer rates. Smokers, and perhaps nonsmoking residents, of smoking households are at increased risk for lung cancer even when radon levels are relatively low. A behavioral risk reduction strategy emphasizing smoking cessation is proposed and data are presented from pilot studies and a short-term evaluation of a randomized intervention trial. METHODS: Pilot studies, including radon testing, interviews, questionnaires, and focus groups, led to a three-arm randomized intervention trial comparing two kinds of written materials and telephone counseling. Smoking households were recruited by offering free radon test kits through an electric utility companies billing system. Three-month follow-up data were obtained by mail and phone. RESULTS: Of an estimated 2,600 smoking households in the utility district, 1,220 requested a radon test kit, and 714 were randomized into three treatment conditions. Brief phone counseling (up to two short calls) significantly increased smoking quit rates, compared to written materials only, and was also related to other risk reduction behaviors (e.g., household ban on smoking). CONCLUSIONS: Offering free radon testing through a public utility billing system is an effective recruitment tool for reaching households at risk due to radon-smoking synergy. Brief telephone counseling is superior to written materials in reducing smoking and encouraging indoor smoking bans. Methods are needed to better inform smokers of their additional risk from exposure to even low levels of radon. PMID- 10479612 TI - Calcium-induced structural changes in synthetic myosin filaments of vertebrate striated muscles. AB - Using negative staining, freeze-drying, and shadowing techniques in electron microscopy we have for the first time demonstrated Ca-induced reversible structural transitions in the synthetic filaments of dephosphorylated column purified rabbit skeletal and cardiac muscle myosins formed by dialysis against solutions containing 120 mM KCl, 1 mM MgCl(2), 10 mM imidazole-HCl buffer (pH 7.0), and either 0.1 mM CaCl(2) or 1 mM EGTA. It has been revealed that the compact ordered structure of the filaments with myosin heads and subfragments-2 (S2) disposed close to the filament backbone with an axial periodicity of about 14.5 nm in the absence of Ca(2+) transforms into a spread disordered structure due to the movement of the heads and S2 away from the filament surface in the presence of Ca(2+). Increasing the pH from neutrality to pH 7.8 leads to a spread, disordered structure while decreasing the pH value to 6.5 returns the filaments to their compact, rather ordered state independent of the Ca(2+) concentrations used. The fact that the reversible structural transitions in synthetic filaments of myosin are observed in the absence of actin and actin- and myosin-associated proteins suggests that Ca(2+)-induced S2 movement is an intrinsic property of myosin itself. Ca(2+)-induced S2 mobility may reflect the existence of functionally significant communications between the myosin head domains and the tails of myosin molecules in thick filaments, and its disappearance can be an indicator of the impairment of these communications, for example, in acute ischemia and myocardial infarction. PMID- 10479613 TI - Modeling alpha-helical coiled coils: analytic relations between parameters. AB - This paper deals with the alpha-helical coiled coil secondary structure of proteins, which is found not only in many fibrous proteins but also in globular proteins. The standard model used nowadays to describe a coiled coil structure is derived from the mathematical description established more than 40 years ago by F. H. C. Crick (1953, Acta Crystallogr. 6, 685-689) from geometrical arguments. In this paper, we apply stereochemical constraints to the protein chains to refine this model. We present a model based on Crick's calculations with less restrictive hypotheses than the standard model and only requiring a set of initial parameters that can be experimentally measured. In addition, the metrics equation method developed here ensures a minimization of the distortions occurring during the coiling process relating the original straight alpha-helix and the coiled coil minor helix. It leads to a modification of the widely used relation between the numbers of residues per turn in the minor and alpha-helices, mathematically demonstrating a previously semiempirical result. This method can be extended to a wide range of coiled structures. PMID- 10479615 TI - Surface processes in the crystallization of turnip yellow mosaic virus visualized by atomic force microscopy. AB - In situ atomic force microscopy (AFM) was used to investigate surface evolution during the growth of single crystals of turnip yellow mosaic virus (TYMV). Growth of the (101) face of TYMV crystals proceeded by two-dimensional nucleation. The molecular structure of the step edges and adsorption of individual virus particles and their aggregates on the crystalline surface were recorded. The surfaces of individual virions within crystals were visualized and seen to be quite distinctive with the hexameric and pentameric capsomers of the T = 3 capsids being clearly resolved. This, so far as we are aware, is the first direct visualization of the capsomere structure of a virus by AFM. In the course of recording the in situ development of the crystals, a profound restructuring of the surface arrangement was observed. This transformation was highly cooperative in nature, but the transitions were unambiguous and readily explicable in terms of an organized loss of classes of virus particles from specific lattice positions. In some cases areas of a single crystal surface were recorded in which were captured successive phases of the transition. We believe this provides the first visual record of a cooperative restructuring of the surface of a supramolecular crystal. PMID- 10479614 TI - Determination of fluorescent probe orientations on biomolecules by conformational searching: algorithm testing and applications to the atomic model of myosin. AB - The ability of a localized conformational searching method to predict probe orientation was tested on model nucleic acid and protein structures and applied to the prediction of skeletal myosin integrity upon chemical modification of its reactive thiols. Double-stranded oligonucleotides were chemically labeled with donor and acceptor resonance energy transfer probes at each end for distance determinations. These measurements were made independently using a terbium chelate as a donor to each of four chemically and spectroscopically distinct acceptor probes from the xanthene and cyanine dye groups. The choice of acceptor significantly affected the separation distance measured. Conformational searching algorithms on the atomic model corrected for the differences to within 0.2 nm on average. Verifying its usefulness on proteins, the localized conformational searching method determined the orientation of a fluorescent probe on RNase A that corresponds closely to available crystallographic models of the labeled protein (RMS deviation = 0.1 nm). Also, analysis of the symmetry of the fluorophores' structures suggests why FRET orientation factors are often closer to their dynamic average value than might normally be expected. Furthermore, the computational method provides insights about FRET data that are important for assessing the stability of the alpha-helix separating the SH1 and SH2 reactive thiols in skeletal myosin. PMID- 10479616 TI - Two-dimensional crystallization on lipid layer: A successful approach for membrane proteins. AB - A considerable interest exists currently in designing innovative strategies to produce two-dimensional crystals of membrane proteins that are amenable to structural analysis by electron crystallography. We have developed a protocol for crystallizing membrane protein that is derived from the classical lipid-layer two dimensional crystallization at the air/water interface used so far for soluble proteins. Lipid derivatized with a Ni(2+)-chelating head group provided a general approach to crystallizing histidine-tagged transmembrane proteins. The processes of protein binding and two-dimensional crystallization were analyzed by electron microscopy, using two prototypic membrane proteins: FhuA, a high-affinity receptor from the outer membrane of Escherichia coli, and the F(0)F(1)-ATP synthase from thermophilic Bacillus PS3. Conditions were found to avoid solubilization of the lipid layer by the detergent present with the purified membrane proteins and thus to allow binding of micellar proteins to the functionalized lipid head groups. After detergent removal using polystyrene beads, membrane sheets of several hundreds of square micrometers were reconstituted at the interface. High protein density in these membrane sheets allowed further formation of planar two-dimensional crystals. We believe that this strategy represents a new promising alternative to conventional dialysis methods for membrane protein 2D crystallization, with the additional advantage of necessitating little purified protein. PMID- 10479617 TI - A two-exposure technique for ice-embedded samples successfully reconstructs the chlorocruorin pigment of Sabella spallanzanii at 2. 1 Nm resolution. AB - A technique for reconstructing ice-embedded macromolecules from electron micrographs taken at two specimen tilts (+/-23 degrees ) has been used to determine the structure of chlorocruorin isolated from the Polychaete annelid Sabella spallanzanii. Images of individual molecules were extracted in couples from two micrographs of the same field of view so each couple consists of two projections of the same molecule. One couple was used as a fixed reference for alignment. Different references yielded reconstructions with different orientations. These were merged to give a model against which the orientation of 1624 first-exposure images was refined to give a final reconstruction at 2.1 nm resolution. The structure of this hematic pigment, essentially the same as that for Lumbricus terrestris, is a bilayer structure with overall symmetry D6, containing six hollow groups per layer. A hollow group is formed by six globular masses and has approximate threefold symmetry. Other structural elements connect the two layers and the hollow groups in a layer. This non-globin material occupies about 15% of the total molecular volume. The results show that the double-exposure strategy, previously described by some of the authors and tested in computer simulations, performs well in real experiments and could be used to obtain preliminary reconstructions in a semiautomatic way. PMID- 10479618 TI - Crystallization of the human, mitochondrial voltage-dependent anion-selective channel in the presence of phospholipids. AB - Overexpressed human voltage-dependent anion-selective channel VDAC or porin from mitochondrial outer membranes has been purified to homogeneity. Electron microscopic analysis of VDAC in detergent solution revealed a uniform particle population consisting of porin monomers. After dialysis of detergent-solubilized porin in the presence of dimyristoylphosphatidylcholine at lipid-to-protein ratios between 0.2 and 0.5 (percentage by weight), mostly multilamellar crystals were obtained. Crystals adsorbed to carbon films flattened during negative staining and air-drying and exhibited different structural features due to differences in the vertical stacking of several crystalline layers, each consisting of one membrane bilayer. Adsorbed, frozen-hydrated multilamellar membrane crystals revealed uniform diffraction patterns with sharp diffraction spots extending to 8.2 A. The surface structure of VDAC was reconstructed from freeze-dried and unidirectionally metal-shadowed crystals. Major protein protrusions were observed from two VDAC monomers present in the unit cell. Differences in the surface structural features indicate alternate orientations of VDAC molecules with respect to the lipid bilayer, allowing the simultaneous imaging of both the cytosolic and intramitochondrial surfaces. Each VDAC molecule consists of a pore lumen with a diameter of 17-20 A surrounded by a protein rim of nonuniform height, suggesting an asymmetrical distribution of protein mass around the diffusion channels. PMID- 10479619 TI - Preliminary crystallographic studies of human mitochondrial NAD(P)(+)-dependent malic enzyme. AB - Human mitochondrial NAD(P)(+)-dependent malic enzyme was overexpressed in Escherichia coli and purified by anion-exchange, ATP affinity, and gel filtration chromatography. The protein was crystallized with the hanging-drop vapor diffusion method. Many different crystal forms were observed, five of which were characterized in some detail. A 2.5-A multiple-wavelength anomalous diffraction data set and a 2.1-A native data set were collected using synchrotron radiation on crystals containing selenomethionyl residues. These crystals belong to space group B2, with a = 204.4 A, b = 107.0 A, c = 59.2 A, and gamma = 101.9 degrees. Self-rotation functions demonstrated that the tetramer of this enzyme obeys 222 symmetry. PMID- 10479620 TI - Crystallization and preliminary X-ray analysis of flavocytochrome c(3), the fumarate reductase from Shewanella frigidimarina. AB - The fumarate reductase (flavocytochrome c(3)) from Shewanella frigidimarina (formerly S. putrefaciens) NCIMB400 has been crystallized in the space group P2(1), with cell dimensions of a = 45.447 A, b = 92.107 A, c = 78.311 A, and beta = 91.038 degrees and one molecule per asymmetric unit. A native data set has been collected to 1.8 A. The gene encoding Fcc(3) from the S. frigidimarina type strain ACAM591 has been cloned and sequenced and the protein crystallized in space group P2(1) with cell dimensions of a = 45.359 A, b = 88.051 A, c = 77.473 A, and beta = 104.499 degrees. Anomalous data have also been collected from the NCIMB400 crystal allowing the heme iron positions to be identified. PMID- 10479621 TI - Purification, crystallization, and preliminary X-ray crystallographic analysis of thermus thermophilus V(1)-ATPase B subunit. AB - The gene of V(1)-ATPase B subunit from the thermophilic eubacterium Thermus thermophilus has been cloned and the protein overproduced in Escherichia coli. The purified protein, with a molecular weight of 53.2 kDa, was crystallized from 10% (w/v) polyethylene glycol 1000, 120 mM magnesium chloride, and 100 mM Na tricine, pH 8.0, by the vapor diffusion method. The crystals diffracted X-rays beyond 3.5 A on a synchrotron radiation source. The crystals belong to the monoclinic space group C2, with unit cell dimensions of a = 153.1 A, b = 129.6 A, c = 92.7 A, and beta = 100.3 degrees. Assuming that three or four molecules are contained in an asymmetric unit, the V(M) value is calculated as 2.8 or 2.1 A (3)/Da, respectively. PMID- 10479622 TI - Crystallization and preliminary diffraction data of 60-kDa glycosylated pollen isoallergens from Bermuda grass. AB - Crystals grown from a 60-kDa isoallergen mixture of Bermuda grass pollen have been obtained in 30% PEG 4000 and 25% isopropanol. The crystals diffract beyond 2 A resolution and belong to a tetragonal space group with the unit cell dimensions a = b = 86 A and c = 310 A. The preferential crystal growth of the larger isoallergens with a blocked N-terminus indicates that crystallization can isolate proteins with compact conformation. PMID- 10479623 TI - Crystallization and preliminary X-ray crystallographic analysis of pyridoxine 5' phosphate oxidase complexed with flavin mononucleotide. AB - Pyridoxine 5'-phosphate oxidase (PNP Ox) catalyzes the terminal step in the biosynthesis of pyridoxal 5'-phosphate. The 53-kDa homodimeric enzyme contains a noncovalently bound flavin mononucleotide (FMN) on each monomer. Three crystal forms of Escherichia coli PNP Ox complexed with FMN have been obtained at room temperature. The first crystal form belongs to trigonal space group P3(1)21 or P3(2)21 with unit cell dimensions a = b = 64.67A, c = 125.64A, and has one molecule of the complex (PNP Ox-FMN) per asymmetric unit. These crystals grow very slowly to their maximum size in about 2 to 4 months and diffract to about 2.3 A. The second crystal form belongs to tetragonal space group P4(1) or P4(3) with unit cell dimensions a = b = 54.92A, c = 167.65A, and has two molecules of the complex per asymmetric unit. The crystals reach their maximum size in about 5 weeks and diffract to 2.8 A. A third crystal form with a rod-like morphology grows faster and slightly larger than the other two forms, but diffracts poorly and could not be characterized by X-ray analysis. The search for heavy-atom derivatives for the first two crystal forms to solve the structure is in progress. PMID- 10479624 TI - Fate of endoleaks after endoluminal repair of abdominal aortic aneurysms with the EVT device. AB - OBJECTIVE: we aim to describe our medium-term follow-up of 20 patients with an endoleak following repair of their abdominal aortic aneurysms (AAA) using the Endovascular Technologies (EVT) device. DESIGN: the experience of one centre in a prospective multicentre phase II trial. MATERIALS AND METHODS: 55 patients with an endovascular repair of their AAA and at least 6 months>> follow-up were reviewed. Intraoperative angiograms, next day duplex scans and computed tomography (CT) images were used to detect endoleaks. Follow-up with CT and duplex was performed at 3, 6, 12 and 24 months. Persistent endoleaks at 6 months were evaluated by angiography and treated by endovascular coiling. RESULTS: there were three immediate conversions to open procedures. Twenty of 52 (38%) patients had an endoleak identified initially. One patient died from a myocardial infarction and three were not evident any longer by discharge CT. Sixteen endoleaks (31%) were present at discharge. Nine resolved spontaneously by 3-6 months and seven were still persistent at 6 months (14%). Six patients underwent coiling of their leak, all with successful radiographic seal after 1-3 sessions. CONCLUSIONS: endoleaks are frequent after endovascular AAA repair, but the majority close spontaneously. Coiling of the leaks and radiographic seal can be achieved in all cases still persistent at 6 months. Whether this method is clinically effective awaits further follow-up. PMID- 10479625 TI - Blood flow in distal end-to-side anastomoses with PTFE and a venous patch: results of an in vitro flow visualisation study. AB - OBJECTIVES: non-physiological flow behaviour plays a significant role in the development of distal anastomotic intimal hyperplasia. To investigate flow patterns in four anastomotic types of femoral end-to-side distal bypass graft anastomoses, a flow visualisation study was performed. METHODS: transparent 1:1 casted replicas of distal vascular graft anastomoses created by conventional technique, Miller-cuff, Taylor- and Linton-patch were fabricated. A pulsatile mock circulation with a high-speed video system was constructed. Flow pattern was determined at mean Reynolds numbers 100-500. Migrations of the stagnation points on the bottom of the anastomoses at mean Reynolds numbers 100, 230, and 350 were measured. RESULTS: a vortex forms during early systole and increases to maximum systole in all anastomoses. During the diastolic phase the vortex moves in the Miller-cuff distally to the toe of the anastomosis and remains standing, while in the other anastomotic types the vortex moves proximally to the heal of the junction and breaks down. The shift of the stagnation point in the Miller-cuff was considerably smaller than in the other anastomoses. CONCLUSION: conventional, Linton and Taylor anastomoses show similar flow patterns. The Miller-cuff with its wider cavity shows lower shift of the bottom stagnation point, but a persistent washout of the anastomotic cavity, which may contribute to its reported good clinical performance. PMID- 10479626 TI - Primary superficial vein reflux with competent saphenous trunk. AB - OBJECTIVES: because reflux in superficial vein tributaries is most often collectively reported with the main saphenous veins, its importance remains largely unrecognised. This study was designed to identify the distribution and extent of non-truncal superficial venous reflux and its association with the signs and symptoms of chronic venous disease (CVD). PATIENTS AND METHODS: eighty four limbs in 62 patients with signs and symptoms of CVD and evidence of reflux on continuous-wave Doppler were subsequently examined with colour-flow duplex imaging. Incompetent superficial vein tributaries were imaged throughout their extent and both ends were identified. Limbs with reflux in the main trunk of the saphenous veins or the deep, perforator or muscular veins, superficial or deep vein thrombosis, injection sclerotherapy, varicose-vein surgery, arterial disease and inflammation of non-venous origin were excluded from the study. The CEAP classification system was used for staging clinical severity of CVD. RESULTS: the prevalence of tributary reflux alone was 9.7% (84/860). Reflux was detected in 171 tributaries. The number of incompetent tributaries ranged from 1 to 5 per limb. Most prevalent were the tributaries to the greater saphenous (111, 65%<0. 0001), followed by those of lesser saphenous (33, 19%) or a combination of both (12, 7%). Incompetent non-saphenous tributaries were uncommon (15, 9%). Among the named tributaries in the lower limb the posterior arch vein was most often incompetent (46, 27%) followed by the anterolateral vein of the thigh (30, 18%), the medial accessory vein (16, 9%) and the anterior arch vein (14, 8%). Reflux in above-the-knee tributaries alone was found in 18 limbs (21%), in below the knee in 23 (28%) and in both sites in 43 (51%). The vast majority of the limbs (71%,p <0.0001) belonged to CVD class 2, 14% in class 3, 9% in class 1 and only 6% in class 4. Class 3 and 4 patients tended to have a longer duration of signs and symptoms, higher number of incompetent tributaries per limb and also a higher prevalence of combined above- and below-knee reflux. CONCLUSIONS: these data indicate that reflux confined to superficial tributaries is found throughout the lower limb. Because this reflux is present without greater and lesser saphenous trunk, perforator and deep-vein incompetence or proximal obstruction, it shows that reflux can develop in any vein without an apparent feeding source. Greater saphenous tributaries are affected significantly more often than those of lesser saphenous, while non-saphenous reflux is uncommon. Most limbs have signs and symptoms of CVD class 2 and 15% belong in classes 3 and 4. PMID- 10479627 TI - Integrated care pathways for vascular surgery. AB - OBJECTIVES: a trial of the use of integrated care pathways (ICPs) for elective vascular surgical procedures. DESIGN: a 12-month prospective study, following a multi-disciplinary group construction of current "best practice" ICPs, with changes in practice only occurring following careful audit of results. MATERIALS: patients admitted to a single vascular unit for "open" repair of abdominal aortic aneurysm, carotid endarterectomy or femoropopliteal bypass grafting. METHODS: patients followed ICPs on a daily basis with signatures required to confirm that action had been taken and careful recording of variances from the ICPs. Audit of variance data allowed changes in the ICPs and, hence, provision of the best possible nursing and clinical practice. RESULTS: ICPs were well received by patients and staff. They improved communication, promoted an appreciation of each health group's role in patient care, increased nursing autonomy, reduced calls to junior medical staff, improved patient education and confidence and caused a marked reduction in hospital "length of stay". CONCLUSIONS: ICPs have clear benefits. This study realises that benefits might be maximal for high throughput, high-cost procedures. Successful use of ICPs depends upon "clinical champions" and effective project management. Sufficient resource and training are essential. PMID- 10479628 TI - Transcranial Doppler monitoring during carotid endarterectomy helps to identify patients at risk of postoperative hyperperfusion. AB - OBJECTIVES: to investigate whether transcranial Doppler (TCD) monitoring can identify patients at risk of hyperperfusion, and whether active postoperative treatment of selected patients decreases the risk of intracerebral haemorrhage (ICH). DESIGN: a case cohort study of 688 patients undergoing carotid endarterectomy (CEA) with intraoperative TCD monitoring. METHODS: sixty-two patients (9%) fulfilled the TCD criteria for hyperperfusion, i.e. >100% increase of peak blood flow velocity or pulsatility index of the middle cerebral artery, compared to preclamp baseline values. In these patients, blood pressure was closely monitored and controlled postoperatively. RESULTS: postoperatively, seven of these patients (11%) exhibited clinical signs or symptoms of hyperperfusion but no cerebral haemorrhage (ICH). This is a significantly better outcome (p <0.005) compared to a 2% incidence of ICH after CEA in previous years in our hospital. CONCLUSIONS: patients at risk of hyperperfusion syndrome after CEA can be identified intraoperatively by TCD monitoring. In these selected patients, immediate and adequate postoperative treatment of hypertension results in a decreased risk of intracerebral haemorrhage. PMID- 10479629 TI - New insights into perforator vein incompetence. AB - BACKGROUND: there has been much controversy on the role of perforator veins in the development of chronic venous disease (CVD). This study was designed to determine the duration and direction of flow of lower limb perforator veins (PVs) in relation to their location, diameter and competency status of superficial and deep veins, in healthy volunteers and patients with different grades of CVD. PATIENTS AND METHODS: thirty limbs in 15 symptom-free volunteers and 103 limbs in 75 patients with signs and symptoms of CVD were examined with colour-flow duplex scanning. Superficial, perforator and deep veins were studied in the standing and sitting positions. Flow-velocity characteristics, the number and maximum PV diameter at the deep fascia and subfascially were determined. A PV was considered incompetent when the outward flow lasted >0.5 s. RESULTS: 581 PVs were found in the patients and 106 in the volunteers. 163 PVs (28%) were incompetent in the first group and none in the latter. The total number of PVs and the number of incompetent PVs per limb increased significantly with the severity of CVD. The mid-calf area had more competent and incompetent PVs in patients (p <0.01). Mean diameter of incompetent PVs in all the CVD classes was significantly larger than that of competent PVs. Competent PVs tended to be larger with increasing severity of CVD and they were significantly larger in the CVD classes 4 to 6 compared to controls (p <0.01). Subfascial PV diameter was markedly larger than that at the fascial level (p <0.001) regardless of the CVD class. A subfascial PV diameter of >3.9 mm (95% CI 3.4 to 4.4 mm) indicated incompetence. However, the reverse was not true, because about a third of incompetent PVs had a subfascial diameter of <3.9 mm. Both competent and incompetent PVs were smaller when located at the lower thigh, knee, ankle and anterior aspect of the calf than those found in the rest of the calf and mid-thigh (p =0. 03). Both inward and outward flow was found more often in patients than in controls (70/418 vs. 9/106, p =0.048). Most incompetent PVs had outward flow alone (126, 77%). PV incompetence was most frequently associated with reflux in superficial veins (120, 74% (p <0.0001), followed by reflux in both the superficial and deep veins (34, 21%) and reflux in the deep veins alone (9, 5%). The mean duration of outward flow was markedly longer in the presence of both superficial and deep vein reflux compared to superficial (p <0.001) or deep vein reflux alone (p <0.0001). CONCLUSIONS: the number of incompetent PVs and the diameter of both competent and incompetent PV increases with the severity of CVD. Bidirectional PV flow is more common in patients than in normal volunteers, while 77% of the incompetent PVs have outward flow alone. PV incompetence is most often associated with reflux in the superficial veins, indicating that deep venous reflux is rarely the primary cause of PV insufficiency. PMID- 10479630 TI - Vascular injuries of the limbs: a fifteen-year Georgian experience. AB - OBJECTIVES: to analyse the causes of injury, surgical approaches, outcome and complication of vascular trauma of the upper and lower limbs in patients with vascular injuries operated on over a period between 1981 and 1995. PATIENTS: in 157 patients, the injuries were penetrating in 136 cases and blunt in 21. Isolated vascular trauma was present in 92 (58.6%) patients, 65 cases (41.4%) were aggrevated by concomitant bone fractures, severe nerve and soft tissue damage. The most frequently injured vessels were the superficial femoral (20. 6%) and brachial (19.1%) arteries. RESULTS: saphenous vein interposition grafting was applied with good results in 34 patients, polytetrafluoroethylene grafts were used in three cases, end-to-end anastomoses in 42 cases, venous bypasses in five cases, and venous patches in seven cases. Seventeen patients underwent arterial repair and nine, venous repair. Fasciotomy was used in 18 cases, and vessels were ligated in 14 cases. Blood flow was restored in 91 patients (58.0%), and collaterals compensated in 31 cases (19.7%). Fourteen primary and nine secondary amputations were performed. Twelve patients died. The limb salvage rate was 77.7% (84.1% among surviving patients). CONCLUSIONS: most vascular injuries associated with limb trauma can be managed successfully unless associated by severe concomitant damage to bones, nerves and soft tissues. PMID- 10479631 TI - Axial flow fields in cuffed end-to-side anastomoses: effect of angle and disease progression. AB - OBJECTIVE: to visualise the axial flow fields in standard and cuffed end-to-side anastomoses (ESA). DESIGN: in vitro experiments using a flow rig, custom-built glass models and frame-by-frame video analysis of flow patterns in standard and cuffed ESA. SUBJECTS: glass models of standard or cuffed (1 cm or 2 cm high) ESA of angles 15, 30, 45 and 60 degrees. RESULTS: the cross-sectional area of standard ESA is much smaller than that of ESA between graft and cuff. The size of the vortex in the anastomotic zone of both standard and cuffed ESA increased with increasing ESA angle and cuff height, but did not change with flow rate. The presence of the vortex maintains a zone of flow separation and low shear at the heel of standard and graft/cuff anastomoses. CONCLUSIONS: the observations explain the clinical findings of intimal hyperplasia (IH) at the heel of PTFE/cuff anastomoses. The improved patency rates of cuffed ESA may be due not to decreased IH, but to an increased ability of the cuff to accommodate IH before causing a significant stenosis. PMID- 10479632 TI - Complications in the endovascular repair of abdominal aortic aneurysms: a risk factor analysis. AB - OBJECTIVES: to identify risk factors for complications following endovascular repair of abdominal aortic aneurysms (AAAs). MATERIALS AND METHODS: endovascular AAA exclusion was attempted in 64 patients. Patient characteristics, anatomic features of the aneurysm, operative technical aspects, and the experience of the teams were correlated with mortality, occurrence of endoleak, and other complications. Perioperative complications were graded following the recommendations of the Ad Hoc Committee on reporting standards. For the assessment of correlation between risk factors and outcomes a logistic regression analysis was used. RESULTS: complications were observed in 43% of the procedures and were classified as mild (24%), moderate (55%) or severe (21%). American Society of Anaesthesiology (ASA) risk class 3 or 4, and advanced age were independent risk factors for perioperative death and complications. Adjuvant procedures or overstenting of the renal arteries with the uncovered part of the stent were not associated with increased risk of complications. Nevertheless, in four of 24 overstented renal orifices, a renal infarction or ischaemia of the kidney was observed on a postoperative CT scan. Advanced experience was associated with less complications, less endoleaks, and shorter operating time. CONCLUSIONS: high age and medical co-morbidity were associated with increased risk for perioperative complications and death. Additional perioperative procedures are usually well tolerated. With greater experience in endovascular AAA grafting the incidence of complications and endoleaks decreased. PMID- 10479633 TI - Below-knee PTFE bypass graft fatigue. PMID- 10479634 TI - Recurrent systemic embolus secondary to free-floating thrombus in the descending thoracic aorta. PMID- 10479635 TI - A soft popliteal mass and pulmonary embolism. PMID- 10479636 TI - A possible side-effect of human erythropoietin therapy: thrombosis of peripheral arterial reconstruction. PMID- 10479642 TI - Hypoxia/Hypoxemia-Induced activation of the procoagulant pathways and the pathogenesis of ischemia-associated thrombosis. AB - Although oxygen deprivation has long been associated with triggering of the procoagulant pathway and venous thrombosis, blood hypoxemia and stasis by themselves do not lead to fibrin formation. A pathway is outlined through which diminished levels of oxygen activate the transcription factor early growth response-1 (Egr-1) leading to de novo transcription/translation of tissue factor in mononuclear phagocytes and smooth muscle cells, which eventuates in vascular fibrin deposition. The procoagulant response is magnified by concomitant suppression of fibrinolysis by hypoxia-mediated upregulation of plasminogen activator inhibitor-1. These data add a new facet to the biology of thrombosis associated with hypoxemia/stasis and imply that interference with mechanisms causing Egr-1 activation in response to oxygen deprivation might prevent vascular fibrin deposition occurring in ischemia without directly interfering with other pro/anticoagulant pathways. PMID- 10479643 TI - Neonatal intima formation in the human coronary artery. AB - Intimal masses develop in the human coronary arteries of all humans, becoming atherosclerotic in later life either because of focal accumulation of lipid or the resulting response to injury. We evaluated the time course of formation of the intimal mass in the proximal left anterior descending coronary artery in autopsy specimens from 91 patients between 17 weeks' gestation and 23 months of postnatal age. Intima was rarely found before 30 weeks' gestation; however, the frequency with which at least some intimal cells were observed increased to 35% between 36 weeks' gestation and birth. By 3 months after birth, all patients had an intimal mass at this coronary location. The mean intima/media ratio was 0.1 just after birth and increased continuously to the second postnatal year. Replication of medial smooth muscle cells, indicated by proliferating cell nuclear antigen staining, was high before birth and decreased between birth and 2 years of age. However, the replication index of the intima remained at 2% to 5%. Thus, coronary intimal cells appearing in the perinatal period may arise by migration after replication of medial smooth muscle, as is seen in models of carotid artery balloon injury. In conclusion, formation of the coronary artery intima is a rapid process, beginning in the peripartum or postpartum period. Given the clonality of the adult lesion and the lack of proliferation in later stages of lesion formation, it is intriguing to speculate that this event may form the basis for atherosclerosis in later life. PMID- 10479644 TI - Angiostatin binds to smooth muscle cells in the coronary artery and inhibits smooth muscle cell proliferation and migration In vitro. AB - Angiostatin is an inhibitor of angiogenesis that is known to reduce endothelial cell proliferation and consequently prevent the progression of tumor metastases. However, the modest effect of angiostatin on endothelial cell proliferation raises the possibility that angiostatin might exert its effects on other cells. To determine the cellular distribution of angiostatin binding in tissues with neovasculature (atherosclerotic coronary arteries), we developed a fusion protein consisting of placental alkaline phosphatase and the first 3 kringles of plasminogen. Angiostatin binding colocalized with smooth muscle cells and could be inhibited by a 50-fold molar excess of plasminogen and 10 mmol/L epsilon-amino n-caproic acid. The fusion protein also bound to smooth muscle cells in culture. Angiostatin inhibited hepatocyte growth factor-induced proliferation and migration of smooth muscle cells, suggesting that they are a target for the antiangiogenic effect of angiostatin. PMID- 10479645 TI - Similarities and differences in smooth muscle alpha-actin induction by TGF-beta in smooth muscle versus non-smooth muscle cells. AB - Transforming growth factor-beta (TGF-beta) has been shown to stimulate smooth muscle (SM) alpha-actin expression in smooth muscle cells (SMCs) and non-SMCs. We previously demonstrated that the 2 CArG boxes A and B and a novel TGF-beta control element (TCE) located within the first 125 bp of the SM alpha-actin promoter were required for TGF-beta inducibility of SM alpha-actin in SMCs. The aims of the present study were (1) to determine whether the TCE exhibits SMC specificity or contributes to TGF-beta induction of SM alpha-actin expression in non-SMCs (ie, endothelial cells and fibroblasts) and (2) to determine whether TGF beta can induce expression of multiple TCE-containing SMC differentiation marker genes, such as SM22alpha, h(1) calponin, and SM myosin heavy chain (SM MHC) in non-SMCs. Results of transient transfection assays demonstrated that mutation of CArG A, CArG B, or the TCE within a 125-bp promoter context completely abolished TGF-beta inducibility of SM alpha-actin in endothelial cells and fibroblasts. However, in contrast to observations in SMCs, inclusion of regions upstream from (-155) completely repressed TGF-beta responsiveness in non-SMCs. Electrophoretic mobility shift assays showed that TGF-beta enhanced binding of a serum response factor to the CArG elements and the binding of an as-yet-unidentified factor to the TCE in endothelial cells and fibroblasts, but to a much lesser extent compared with SMCs. TGF-beta also stimulated expression of the SMC differentiation marker SM22alpha in non-SMCs. However, in contrast to SMCs, TGF beta did not induce expression of h(1) calponin and SM MHC in non-SMCs. In summary, these results suggest a conserved role for CArG A, CArG B, and the TCE in TGF-beta-induced expression of SM alpha-actin in SMCs and non-SMCs that is modified by a complex interplay of positive- and negative-acting cis elements in a cell-specific manner. Furthermore, observations that TGF-beta stimulated expression of several early but not late differentiation markers in non-SMCs indicate that TGF-beta alone is not sufficient to induce transdifferentiation of non-SMCs into SMCs. PMID- 10479646 TI - Estrogen mediates the protective effects of pregnancy and chorionic gonadotropin in a mouse model of vascular injury. AB - To determine why pregnancy protects against intimal proliferation in a mouse model of vessel injury, we administered chorionic gonadotropin to intact and ovariectomized female mice. Chorionic gonadotropin markedly suppressed intimal proliferation in intact but not in ovariectomized female mice, indicating that the protective effects of chorionic gonadotropin require ovarian function. To test whether estrogen or progesterone might mediate the protective effects of pregnancy and chorionic gonadotropin, we administered estrogen and progesterone to ovariectomized mice. Estrogen administration to ovariectomized mice to achieve the elevated levels seen in pregnancy was sufficient to reproduce the marked suppression of intimal proliferation in response to vessel injury. Progesterone administration reduced intimal proliferation to a lesser degree and was correlated with increases in estrogen to levels seen in nonpregnant female mice. Staining for proliferating cell nuclear antigen suggested that estrogen reduced medial and intimal cell proliferation. Both the classic estrogen receptor-alpha and the recently discovered estrogen receptor-beta are present in vascular tissue as assessed by immunohistochemistry, providing a possible mechanism for the effects of estrogen. These results suggest that the protective effects of estrogen do not plateau at levels seen in normal females but increase further with estrogen levels up through levels seen during pregnancy. PMID- 10479647 TI - Apoptosis and overexpression of bax protein and bax mRNA in smooth muscle cells within intimal hyperplasia of human radial arteries : analysis with arteriovenous fistulas used for hemodialysis. AB - There is a type of arteriosclerosis with remodeling of middle-size arteries in which intimal hyperplasia of smooth muscle cells (SMCs) plays the main role, and there are few macrophages, T lymphocytes, and foam cells. It is unknown whether apoptosis and the expression of Bax, an inducer of apoptosis, are increased according to the progression of this type of human arteriosclerosis, which is different from so-called atherosclerosis. Bax heterodimerizes with Bcl-2, an inhibitor of apoptosis, and the ratio of Bax to Bcl-2 determines cellular apoptosis or survival. Thus, we investigated apoptosis and the expressions of Bax, bax mRNA, and Bcl-2 in human arteriovenous (AV) fistulas used for hemodialysis, a representative of arteriosclerosis of the aforementioned type. The material was 20 radial arteries obtained from 20 patients with chronic renal failure undergoing AV shunt surgery. SMCs, macrophages, and T lymphocytes were immunohistochemically identified at the light microscopic (LM) level. Apoptosis was detected by in situ terminal deoxynucleotidyl transferase (TdT)-mediated digoxigenin-dUTP nick end labeling (TUNEL) at both the LM and electron microscopic (EM) level. Cell proliferating activity was estimated by proliferating cell nuclear antigen (PCNA). Bax and Bcl-2 were detected by immunohistochemistry and Western blot analysis. Expression of bax mRNA was detected by in situ hybridization. LM TUNEL-positive cells in both the intima and media were significantly increased according to the percent stenosis of the vessels. EM analysis revealed that ultrastructures of apoptotic SMCs were seen in both synthetic and contractile phenotypes. Their frequency of occurrence in the intima and media were greater in those vessels with >50% stenosis than in those with <50% stenosis (5.2+/-0.7% versus 1.0+/-0.3% in the intima and 2. 1+/-0.5% versus 0.2+/-0.1% in the media). The proportion of apoptotic SMCs with ruptured plasma membranes was greater than that of apoptotic SMCs with intact membranes in the intima of the former (4.1+/-0.6% versus 1.1+/-0.1%). Only those SMCs with apoptotic ultrastructures had TUNEL-positive nuclei with moderate or marked accumulation of immunogold particles at the EM level. However, ultrastructures of oncosis (primary necrosis) were not observed. Immunohistochemical analyses showed significant positive correlations between percent stenosis of vessels and the percentage of either PCNA-positive intimal cells or Bax-positive areas in the intima and media. Bcl-2-positive cells were not observed in the intima but mainly in the outer media. The percentage of Bcl-2-positive medial cells was definitely decreased at an early stage after formation of the AV fistula but did not change according to the duration of hemodialysis or the progression of arteriosclerosis. Western blot analysis of Bax or Bcl-2 and in situ hybridization of bax mRNA confirmed the immunohistochemical data. Thus, regulation of cellularity in intimal hyperplasia of SMCs in human arteriosclerosis with remodeling is mediated by proliferation and apoptosis but not oncosis. The apoptosis is probably induced by an increase in the Bax to Bcl-2 ratio. PMID- 10479648 TI - Adenovirus-mediated overexpression of c-Jun and c-Fos induces intercellular adhesion molecule-1 and monocyte chemoattractant protein-1 in human endothelial cells. AB - As distal targets and mediators of signal transduction pathways, activator protein-1 (AP-1), c-Jun, and c-Fos are among the primary regulators of genes involved in cell function, proliferation, and differentiation. By using adenovirus-mediated gene transfer, we show that overexpression of AP-1 proteins directly causes coinduction of gene expression of an adhesion molecule, intercellular adhesion molecule-1 (ICAM-1), and a chemokine, monocyte chemoattractant protein-1 (MCP-1), in human vascular endothelial cells (ECs). The AP-1-induced gene expression occurs through a mechanism independent of nuclear factor-kappaB. Because the induced expression of ICAM-1 and MCP-1 in ECs has been implicated in endothelial activation and a number of important vascular disorders, it is suggested that AP-1 activation may play an important role in the pathogeneses of inflammation, angiogenesis, and atherogenesis. PMID- 10479649 TI - Expression of CCR2 by endothelial cells : implications for MCP-1 mediated wound injury repair and In vivo inflammatory activation of endothelium. AB - Endothelial cell proliferation and migration may play a central role in angiogenesis, wound healing, and atherosclerosis. Although CXC chemokines can act on endothelial cells by influencing proliferation, an involvement of CC chemokines and endothelial expression of chemokine receptors remains to be elucidated. Reverse transcription-polymerase chain reaction, RNase protection, Western blot, and flow cytometric analysis showed that human umbilical vein endothelial cells express mRNA and surface protein of the monocyte chemotactic protein-1 (MCP-1) receptor CCR2, which was upregulated by inflammatory cytokines. MCP-1 induced migration of endothelial cells in a transwell assay, which was inhibited by the 9-76 MCP-1 receptor antagonist. Increased secretion of MCP-1 or interleukin-8, but not RANTES, on endothelial injury suggested a functional role of CCR2 in wound repair as measured by ELISA. After mechanical injury to endothelial monolayers, which spontaneously closed within 24 hours, wound repair was delayed by the 9-76 antagonist and by a blocking monoclonal antibody to MCP 1, but not to interleukin-8, and was improved by exogenous MCP-1. This was confirmed by quantification of cell migration into the wound area, whereas proliferation and viability were unaltered by MCP-1 or its analogue. Notably, immunohistochemistry of inflamed tissue revealed CCR2 staining on arterial, venous, and venular endothelium affected by cellular infiltration. This is the first demonstration of endothelial CCR2 expression ex vivo, inferring its involvement in inflammatory conditions. Thus endothelial cells express functional CCR2 that may have important implications for endothelial wound repair and inflammatory reactions. PMID- 10479650 TI - Peroxisome proliferator-activated receptor activators target human endothelial cells to inhibit leukocyte-endothelial cell interaction. AB - An early event in acute and chronic inflammation and associated diseases such as atherosclerosis and rheumatoid arthritis is the induced expression of specific adhesion molecules on the surface of endothelial cells (ECs), which subsequently bind leukocytes. Peroxisome proliferator-activated receptors (PPARs), members of the nuclear receptor superfamily of transcription factors, are activated by fatty acid metabolites, peroxisome proliferators, and thiazolidinediones and are now recognized as important mediators in the inflammatory response. Whether PPAR activators influence the inflammatory responses of ECs is unknown. We show that the PPAR activators 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), Wyeth 14643, ciglitazone, and troglitazone, but not BRL 49653, partially inhibit the induced expression of vascular cell adhesion molecule-1 (VCAM-1), as measured by ELISA, and monocyte binding to human aortic endothelial cells (HAECs) activated by phorbol 12-myristate 13-acetate (PMA) or lipopolysaccharide. The "natural" PPAR activator 15d-PGJ(2) had the greatest potency and was the only tested molecule capable of partially inhibiting the induced expression of E-selectin and neutrophil-like HL60 cell binding to PMA-activated HAECs. Intracellular adhesion molecule-1 induction by PMA was unaffected by any of the molecules tested. Both PPAR-alpha and PPAR-gamma mRNAs were detected in HAECs by using reverse transcription-polymerase chain reaction and a ribonuclease protection assay; however, we have yet to determine which, if any, of the PPARs are mediating this process. These results suggest that certain PPAR activators may help limit chronic inflammation mediated by VCAM-1 and monocytes without affecting acute inflammation mediated by E-selectin and neutrophil binding. PMID- 10479651 TI - 9-cis retinoic acid induces monocyte chemoattractant protein-1 secretion in human monocytic THP-1 cells. AB - Monocyte migration and activation are regulated by monocyte chemoattractant protein-1 (MCP-1). Prior studies have shown MCP-1 expression is modulated by a variety of ligands that act through extracellular receptors. In the current study, we show 9-cis retinoic acid (RA), a ligand for the nuclear hormone receptor retinoid X receptor (RXR) and retinoic acid receptor (RAR), markedly induces the expression of MCP-1. In human THP-1 monocytic leukemia cells cultured with RA (0.05 to 500 nmol/L), MCP-1 expression was induced rapidly, significantly, and dose-dependently by as much as 165-fold. MCP-1 RNA level was also increased in RA-treated cells. Expression of PPARgamma, a heterodimer partner of RXR, is also markedly induced by RA in THP-1 cells. However, BRL49653, a PPARgamma ligand, failed to induce MCP-1 secretion either alone or to modify the expression level induced by RA. In contrast, BRL49653 significantly increased MCP-1 (biotinylated MCP-1) binding to THP-1 cells, whereas RA had no effect. Other peroxisome proliferator activated receptor (PPAR) ligands, 15d-PGJ(2) and troglitazone (PPARgamma), Wy14,643 (PPARalpha), and PD195599 (PPARbeta) inhibited the induction of MCP-1 by RA. RA's effect on MCP-1 expression in human elutriated monocytes were similar to that observed in the THP-1 cells. These studies identify RA as a nuclear signal for MCP-1 induction in undifferentiated human monocytic cells. These studies also suggest monocyte MCP-1 expression induced through RA may modulate cell migration. PMID- 10479652 TI - Dexamethasone enhances In vitro vascular calcification by promoting osteoblastic differentiation of vascular smooth muscle cells. AB - Vascular calcification is often associated with atherosclerotic lesions. Moreover, the process of atherosclerotic calcification has several features similar to the mineralization of skeletal tissue. Therefore, we hypothesized that vascular smooth muscle cells might acquire osteoblastic characteristics during the development of atherosclerotic lesions. In the present study, we investigated the effect of dexamethasone (Dex), which is well known to be a potent stimulator of osteoblastic differentiation in vitro, on vascular calcification by using an in vitro calcification model. We demonstrated that Dex increased bovine vascular smooth muscle cell (BVSMC) calcification in a dose- and time-dependent manner. Dex also enhanced several phenotypic markers of osteoblasts, such as alkaline phosphatase activity, procollagen type I carboxy-terminal peptide production, and cAMP responses to parathyroid hormone in BVSMCs. We also examined the effects of Dex on human osteoblast-like (Saos-2) cells and compared its effects on BVSMCs and Saos-2 cells. The effects of Dex on alkaline phosphatase activity and the cAMP response to parathyroid hormone in BVSMCs were less prominent than those in Saos-2 cells. Interestingly, we detected that Osf2/Cbfa1, a key transcription factor in osteoblastic differentiation, was expressed in both BVSMCs and Saos-2 cells and that Dex increased the gene expression of both transcription factors. These findings suggest that Dex may enhance osteoblastic differentiation of BVSMCs in vitro. PMID- 10479653 TI - Angiotensin II activation of insulin-like growth factor 1 receptor transcription is mediated by a tyrosine kinase-dependent redox-sensitive mechanism. AB - We have recently shown that angiotensin II activation of insulin-like growth factor 1 receptor (IGF-1R) transcription is a critical requirement for angiotensin-stimulated vascular smooth muscle cell growth; therefore, we examined the signaling pathway involved. In rat aortic smooth muscle cells, the antioxidants N-acetyl-L-cysteine (5 mmol/L) and pyrrolidine dithiocarbamate (100 micromol/L) completely inhibited angiotensin II-stimulated increases in IGF-1R mRNA and protein levels, suggesting the involvement of reactive oxygen species. Indeed, catalase abolished the Ang II-stimulated increase of IGF-1R protein expression, and accordingly, H(2)O(2) (0.2 mmol/L) or the oxidized products of linoleic acid, hydroperoxyoctadecadienoic acids (10 micromol/L), increased IGF-1R mRNA levels at 3 hours by 74+/-20% and 107+/-22% and increased receptor number at 24 hours by 51+/-6.7% and 55+/-7.4%, respectively. The protein tyrosine kinase inhibitors genistein and tyrphostin A25 also blocked angiotensin II increases in IGF-1R mRNA and protein levels and blocked the ability of hydroperoxyoctadecadienoic acids and H(2)O(2) to increase IGF-1R expression, suggesting that oxidative stress may be an early event in the angiotensin II signaling cascade. Furthermore, calcium chelation inhibited the angiotensin II effect. Transient transfection assays revealed that a (-2350)+640 IGF-1R promoter/luciferase construct was fully responsive to angiotensin II stimulation (127+/-20% increase). Ten millimoles per liter hydroperoxyoctadecadienoic acids and 0.2 mmol/L H(2)O(2) increased luciferase activity by 79+/-8.5% and 63+/-12%, respectively, and 5 mmol/L N-acetyl-L-cysteine blocked the angiotensin II-induced upregulation of luciferase activity by 70%. These data suggest that angiotensin II stimulates IGF-1R gene transcription via calcium-dependent activation of protein tyrosine kinase activity that lies downstream from an oxidant stimulus. These findings provide key insights into the signaling mechanisms whereby angiotensin II exerts its growth-promoting effects on the vasculature. PMID- 10479654 TI - Phosphatidylinositol 3-kinase is required for growth factor-induced amino acid uptake by vascular smooth muscle cells. AB - Although accumulating evidence suggests that phosphatidylinositol 3-kinase (PI3K) is a common signaling molecule for growth factor-induced amino acid uptake by the cell, the role of PI3K in the uptake of different amino acids was not tested under the same conditions. In this study, we asked whether PI3K mediates platelet derived growth factor (PDGF) -stimulated uptake of different amino acids that are taken up through 3 major amino acid transporters expressed in rat vascular smooth muscle cells and other cell types and whether PI3K mediates amino acid uptake stimulated with different growth factors and vasoactive substances. PDGF increased the uptake of [(3)H]leucine, [(3)H]proline, and [(3)H]arginine in a dose- and time-dependent fashion. Two different PI3K inhibitors, wortmannin (100 nmol/L) and LY294002 (10 micromol/L), completely inhibited the amino acid uptake stimulated by PDGF. Chinese hamster ovary cells expressing both PDGF receptor beta and a dominant-negative PI3K did not increase their leucine uptake when stimulated with PDGF, whereas the same cells expressing only PDGF receptor-beta did. Transforming growth factor-beta, as well as insulin-like growth factor-I and angiotensin II, increased leucine uptake by vascular smooth muscle cells. Wortmannin and LY294002 inhibited this increase. We also found that transforming growth factor-beta stimulated PI3K activity and the phosphorylation of Akt, a downstream signaling molecule of PI3K. A similar effect of PI3K inhibitors on amino acid uptake was observed in Swiss 3T3 cells. We conclude that PI3K mediates the uptake of different amino acids by vascular smooth muscle cells and other cell types stimulated with a variety of growth factors, including transforming growth factor-beta. Our findings suggest that PI3K may play an important role in vascular pathophysiology by regulating amino acid uptake. PMID- 10479655 TI - Phosphatidylinositol 3-kinase and focal adhesion kinase are early signals in the growth factor-like responses to thrombospondin-1 seen in human vascular smooth muscle. AB - Thrombospondin-1 (TSP-1) is a matricellular protein that is expressed in negligible amounts in normal blood vessels but is markedly upregulated in vascular injury. Although TSP-1 can act as a pleiotropic regulator for human vascular smooth muscle cells (HVSMCs), the intracellular signaling pathways stimulated by this protein remain obscure. In cultured HVSMCs derived from saphenous vein, TSP-1 induces tyrosine phosphorylation of a number of cellular proteins, with a complex temporal pattern of activation. Immunoprecipitation techniques have identified the early tyrosine-phosphorylated signals as being the p85 regulatory subunit of phosphatidylinositol 3-kinase (PI 3-K) and focal adhesion kinase (FAK). Tyrosine phosphorylation of the p85 subunit of PI 3-K showed a biphasic response to TSP-1 stimulation, which corresponded to a biphasic activation of the lipid kinase. Treatment with both wortmannin and LY294002 inhibited PI 3-K activity of HVSMCs but did not affect tyrosine phosphorylation of the p85 regulatory subunit. TSP-1-stimulated FAK phosphorylation, however, was substantially reduced by these inhibitors, as was the TSP-1-induced chemotaxis of these cells. These results suggest that activation of PI 3-K is an early signal induced by TSP-1 and is critical for chemotaxis. Activation of this kinase precedes and may occur upstream from FAK phosphorylation, although the nature of the interaction between these 2 enzymes remains obscure. PMID- 10479656 TI - Selective activation of the prostanoid EP(3) receptor reduces myocardial infarct size in rodents. AB - The cardioprotective effects of E-type prostaglandins (EPs) have been attributed to vasodilatation, inhibition of platelet and neutrophil function (EP(2) mediated), and an unknown "cytoprotective effect." We have hypothesized that selective activation of EP(3) receptors may cause cardioprotection. The prostanoid derivative ONO-AE-248 selectively binds to murine EP(3alpha) receptors expressed in Chinese hamster ovary (CHO) cells (K(i), 15 nmol/L) and prevents the rise in cAMP caused by forskolin in CHO cells (IC(50) approximately 1 nmol/L) in which the EP(3alpha) receptor had been expressed. In anesthetized rats subjected to regional myocardial ischemia for 25 or 45 minutes and 2 hours of reperfusion, infusion of ONO-AE-248 (5 microg kg(-1). min(-1) IV) caused a significant reduction in infarct size, from 60+/-3% (n=8) to 36+/-6% (n=7) and from 78+/-2% (n=11) to 58+/-4% (n=9), respectively. The reduction in infarct size caused by ONO-AE-248 in rats subjected to 25 minutes of ischemia and reperfusion was abolished by a selective inhibitor of ATP-sensitive potassium (K(ATP)) channels, 5-hydroxydecanoate (n=6), and the protein kinase C inhibitors staurosporine (n=6) and chelerythrine (n=6). In anesthetized rabbits subjected to coronary artery occlusion for 45 or 60 minutes and 2 hours of reperfusion, infusion of ONO-AE-248 (5 microg. kg(-1). min(-1) IV) caused a significant reduction in infarct size, from 61+/-2% (n=10) to 36+/-4% (n=8) and from 63+/-4% (n=7) to 42+/-4% (n=7), respectively. The reduction in infarct size caused by ONO-AE-248 in the rabbit was also abolished by 5-hydroxydecanoate. The cardioprotective effect of ONO-AE 248 in rats or rabbits was not associated with any hemodynamic effects. Selective activation of the prostanoid EP(3) receptor reduces myocardial infarct size in rodents by a mechanism(s) that may involve the activation of protein kinase C and the opening of K(ATP) channels. PMID- 10479657 TI - Chronic endothelin-1 improves nitric oxide-dependent flow-induced dilation in resistance arteries from normotensive and hypertensive rats. AB - Endothelin-1 (ET-1) is released on stimulation by shear stress of the vascular wall. In several pathological situations, an involvement of ET-1 is suspected. Nevertheless, the effect of a chronic increase in circulating ET-1 on vascular tone in resistance arteries is not yet fully understood. We investigated the response to tensile stress (pressure-induced myogenic tone) and shear stress (flow-induced dilation, FD) of rat mesenteric resistance arteries cannulated in an arteriograph. Intraluminal diameter was measured continuously. Rats (normotensive Wistar-Kyoto rats [WKYs] and spontaneously hypertensive rats [SHRs]) were treated for 2 weeks with ET-1 (5 pmol. kg(-1). min(-1) SC; n=8 to 16 per group). Systolic arterial blood pressure increased significantly in ET-1 treated rats (171+/-7 versus 196+/-6 mm Hg in WKYs and 216+/-8 versus 245+/-6 mm Hg in SHRs, P<0.05). Passive arterial diameter in isolated resistance arteries ranged from 78+/-9 to 169+/-4 microm in WKYs and from 62+/-6 to 149+/-7 microm in SHRs (pressure from 10 to 150 mm Hg). Myogenic tone was not significantly affected by chronic ET-1. Flow (9 to 150 microL/min) significantly increased the arterial diameter by 2+/-0.5 to 22+/-2 microm in WKYs and by 1.3+/-0. 7 to 8.3+/ 0.8 microm in SHRs (P<0.001 versus WKYs). The NO synthesis blocker N(G)-nitro-L arginine methyl ester (L-NAME; 100 micromol/L) attenuated FD in WKYs (eg, 22+/-2 versus 15+/-3 microm after L-NAME, flow=150 microL/min) and, to a lesser extent, in SHRs (P<0.001 versus WKYs). The cyclooxygenase inhibitor indomethacin (3 micromol/L) attenuated the remaining FD in WKYs (eg, 15+/-3 versus 8+/-3 microm, flow=150 microL/min) and in SHRs (eg, 7.5+/-0.5 versus 5.0+/-0.6 microm). Chronic ET-1 significantly increased FD in SHRs but not in WKYs. In both strains, NO dependent FD was significantly increased by chronic ET-1. Furthermore, indomethacin-sensitive FD was increased by chronic ET-1 in SHRs only. Thus, chronic ET-1 increased NO-dependent FD in resistance mesenteric arteries from both WKYs and SHRs and increased indomethacin-sensitive FD in SHRs only. PMID- 10479658 TI - Different effects of photodynamic therapy and gamma-irradiation on vascular smooth muscle cells and matrix : implications for inhibiting restenosis. AB - gamma-Irradiation (gamma-RT) and photodynamic therapy (PDT) are known to inhibit intimal hyperplasia. The common mechanism is that both modalities produce free radicals, but unlike gamma-RT, PDT generates them through the absorption of light by photosensitizers. The purpose of this in vitro study was to assess the differences that PDT and gamma-RT have on the fibroproliferative response after vascular injury by comparing their effects on vascular smooth muscle cells (SMCs) and on the extracellular matrix (ECM). Mitochondrial activity (tetrazolium salt), proliferation ([(3)H]thymidine incorporation), and the mechanisms of cell death (terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling [TUNEL] staining) were used to assess differences between PDT (100 J/cm(2)) and gamma-RT (10 or 20 Gy) on SMC injury. The different effects on bioregulatory molecules were investigated by quantitating the proliferation of SMCs cultured with conditioned medium and on treated ECM. PDT of SMCs reduced proliferation and mitochondrial activity (0.5+/-0.75% and 1.7+/-4.25%, respectively, P<0.0001), whereas gamma-RT of SMCs decreased cell proliferation but did not affect metabolic activity. Stimulation with calf serum of gamma-RT-treated SMCs did not affect proliferation but increased mitochondrial enzyme activity (160+/-11%, P<0.0005). The conditioned medium, derived from PDT- but not gamma-RT-treated SMCs, did not stimulate effector SMC proliferation compared with gamma-RT-treated SMCs (16+/-4.1% versus 80+/-16.8%, P<0.0001). Apoptosis was the principle cytotoxic mechanism after PDT, whereas gamma-RT cells were growth arrested but viable. PDT of the ECM reduced effector SMC proliferation compared with controls and gamma-RT cells (18+/-6.5% versus 100+/-17.7% and 84+/-8.9%, respectively, P<0.0001). These data suggest that gamma-RT and PDT may inhibit restenosis but by different mechanisms. The effects of PDT are more diverse and may result in improved outcome while avoiding the teratogenic exposure due to ionizing irradiation. PMID- 10479659 TI - Rapid regression of atherosclerosis induced by liver-directed gene transfer of ApoE in ApoE-deficient mice. AB - Apolipoprotein E (apoE) is a multifunctional protein synthesized by the liver and tissue macrophages. ApoE-deficient mice have severe hyperlipidemia and develop accelerated atherosclerosis on a chow diet. Both liver-derived and macrophage derived apoEs have been shown to reduce plasma lipoprotein levels and slow the progression of atherosclerosis in apoE-deficient mice, but regression of atherosclerosis has not been demonstrated in this model. We utilized second generation adenoviruses to achieve hepatic expression of human apoE in chow-fed, apoE-deficient mice with established atherosclerotic lesions of different stages. As expected, hepatic expression of human apoE3 significantly reduced plasma cholesterol levels. Liver-derived apoE also accumulated substantially within preexisting atherosclerotic lesions, indicating that plasma apoE gained access to the arterial intima. Hepatic expression of human apoE3 for 6 weeks resulted in significant quantitative regression of both early fatty streak lesions as well as advanced, complex lesions in both the aortic root and the aortic arch. In addition, hepatic expression of apoE induced substantial morphological changes in lesions, including decreased foam cells and increased smooth muscle cells and extracellular matrix content. In parallel, human apoE4 and apoE2 were also expressed in the liver by using recombinant adenoviruses. ApoE4 reduced cholesterol levels to the same extent as did apoE3 and also prevented progression but did not induce significant regression of preexisting lesions. ApoE2 reduced cholesterol levels to a lesser degree than did apoE3 and apoE4 and lesion progression was reduced, but regression was not induced. In summary, (1) regression of preexisting atherosclerotic lesions in apoE-deficient mice can be rapidly induced by hepatic expression of apoE, despite the absence of macrophage derived apoE; (2) the morphological changes seen in this model of regression resemble those in other animal models, induced over longer periods of time; (3) liver-derived apoE gained access to and was retained by intimal atherosclerotic lesions; and (4) apoE4 was less effective in inducing regression, despite its effects on plasma lipoproteins that were similar to those of apoE3. The rapid regression of preexisiting atherosclerotic lesions induced by apoE gene transfer in apoE-deficient mice could provide a convenient murine model for investigation of the molecular events associated with atherosclerosis regression. PMID- 10479660 TI - Local hypomethylation in atherosclerosis found in rabbit ec-sod gene. AB - Extracellular superoxide dismutase (EC-SOD) protects arteries against deleterious effects of superoxide anions and the development of atherosclerosis. In this study, we cloned and characterized rabbit ec-sod gene. We identified 6 rabbit C elements and 5 CpG clusters in the cloned sequence. One of the CpG clusters is located on the coding sequence. Because CpG clusters are potential sites for methylation and may explain the occurrence of mutations, methylation status of each of the CpG dimers located in the coding sequence CpG cluster was characterized using direct genomic sequencing. Unexpectedly, a marked reduction in the amount of methylated CpG dinucleotides in ec-sod gene was detected in atherosclerotic aortas as compared with normal aortic intima-media. Although alterations in DNA methylation are well characterized in malignant tumors, the presence of methylation changes in atherosclerosis has not been studied even though both diseases are characterized by excess cellular proliferation and alterations in gene expression. Further analysis of the whole genomic methylation by high-pressure liquid chromatography in normal and atherosclerotic aortas revealed a tendency for a decreased 5-methylcytosine (5-mC) content in atherosclerotic aortas as compared with normal arteries. Hypomethylation in atherosclerotic aortas occurred at the same level as has been reported from malignant tumors. Although a causal relationship between the methylation level and expression of EC-SOD cannot be proven, our results show that ec-sod hypomethylation is associated with the development of atherosclerosis and suggest that it may affect structure and function of ec-sod and other genes possibly involved in the development of atherosclerotic lesions. PMID- 10479661 TI - Cholesterol oxidation products induce vascular foam cell lesion formation in hypercholesterolemic New Zealand white rabbits. AB - Circulating cholesterol oxidation products (ChOx) have long been implicated in the etiology of early atherosclerosis; however, direct in vivo evidence elucidating their role in atherogenesis is only recently becoming available. This study investigated ChOx effects on vascular lesion formation in New Zealand White rabbits under controlled hypercholesterolemic conditions. By closely monitoring plasma cholesterol levels and adjusting dietary cholesterol intake during a 78 day period, total plasma cholesterol exposures (cumulative plasma cholesterol levels over time) were controlled between 27 000 and 34 000 mg/dLxday (final plasma cholesterol concentration, 467+/-77 mg/mL), representing a threshold range for sudanophilic lesion formation in the aorta. Twenty injections of a ChOx mixture (70 mg per injection) were made bearing an oxysterol composition similar to that found in circulating oxidatively modified low density lipoprotein. At sacrifice, the ChOx-injected rabbits (n=5) had (1) significantly higher plasma ChOx levels, (2) significantly increased cholesterol content in the aortas, mainly as esterified cholesterol, and (3) significantly greater sudanophilic lesion size and frequency in the aortas compared with vehicle-injected control rabbits (n=5). The aortic cholesterol content and extent of sudanophilic lesion area were correlated significantly with total plasma ChOx exposure (P<0.003 and P<0.0001, respectively) but not with total cholesterol exposure. The results indicate that for moderate experimental hypercholesterolemia, a situation more relevant to physiological hypercholesterolemia in humans, circulating ChOx may play an important role in inducing formation of early atherosclerotic lesions. Because ChOx are often present in cholesterol-containing diets, foam cell lesion formation induced by ChOx rather than cholesterol cannot be overlooked. PMID- 10479662 TI - Coronary and aortic calcification among women 8 years after menopause and their premenopausal risk factors : the healthy women study. AB - In the Healthy Women Study, the relationship between cardiovascular risk factors measured premenopausally at age 48, use of hormone therapy, and coronary and aortic calcification at age 58 were evaluated among 169 women. Approximately 63% of women had no coronary calcification, but only 29% had no aortic calcification. Coronary calcification and aortic calcification were positively correlated with each other. There was a very strong association between low density lipoprotein cholesterol (LDL-C) level and coronary calcification. Among women with premenopausal levels of LDL-C <100 mg/dL, only 9% had a coronary calcium score >/=101 compared with 30% of women with an LDL-C >160 mg/dL. Only 5% of women with a high density lipoprotein cholesterol (HDL-C) level >60 mg/dL had high coronary scores. The level of HDL(2)-C was especially strongly inversely related to coronary calcium scores. Cigarette smoking was a very important determinant of both high aortic and high coronary calcium scores. Other risk factors associated with greater coronary calcium were higher systolic blood pressure, triglycerides levels, and blood glucose. Use of hormone replacement therapy was associated with less coronary calcium (NS). For both hormone replacement therapy users and nonusers, the levels of LDL-C and HDL-C measured premenopausally were predictors of coronary and aortic calcium scores. Thus, risk factors evaluated premenopausally are powerful predictors of coronary and aortic calcification, a marker of atherosclerosis, measured 8 years after menopause, 11 years later in these women. PMID- 10479663 TI - Novel effects of the acyl-coenzyme A:Cholesterol acyltransferase inhibitor 58-035 on foam cell development in primary human monocyte-derived macrophages. AB - We examined the effect of acyl-coenzyme A:cholesterol acyltransferase (ACAT) inhibitors on intracellular cholesterol stores in primary human monocyte-derived macrophages (HMMs) during foam cell formation. HMMs were exposed to acetylated low density lipoprotein (acLDL, 500 microg protein per mL) with or without 58-035 (1 to 10 microg/mL) or CI-976 (2 microg/mL) for 2 to 48 hours. Total cholesterol (TC) and esterified cholesterol (EC) mass was significantly lower while unesterified cholesterol (UC) increased slightly in cells incubated with acLDL plus ACAT inhibitors. Sterol mass was also measured in cells coincubated with acLDL (500 microg protein per mL) with or without 58-035 (2 microg/mL), high density lipoprotein (HDL, 400 microg protein per mL), or HDL+58-035 for 48 hours. TC and EC were 23% and 55% lower, respectively (P<0.0004), while UC was 11% higher (P<0.04) in cells incubated with acLDL plus 58-035. In contrast, coincubation with HDL alone did not significantly affect TC, EC, or UC mass compared with acLDL alone. The effect of 58-035 could not be explained by cytotoxicity, because adenine release, secreted lactate dehydrogenase, glucose utilization, and cell protein were similar in cells exposed to acLDL regardless of the presence of 58-035. We investigated several potential mechanisms for the decreased TC mass, including increased UC efflux and decreased acLDL binding and uptake. Efflux was measured in cells exposed to [1,2-(3)H]cholesteryl oleate labeled acLDL, unlabeled control acLDL, and native untreated acLDL (500 microg protein per mL) with or without 58-035 (5 microg/mL) for 24 or 48 hours. UC efflux increased in a time-dependent manner from cells exposed to acLDL plus 58 035 compared with cells exposed to acLDL alone (P<0. 04). High-affinity binding was measured in cells exposed to (125)I-acLDL (5 microg protein per mL) with or without excess unlabeled acLDL (100 or 500 microg protein per mL) for 4 hours at 4 degrees C. Specific acLDL binding, uptake, and total degradation were significantly lower when 58-035 was present during cholesterol enrichment compared with cells exposed to acLDL alone (P<0.001). Unlike the effects of ACAT inhibitors on foam cell formation in rodent macrophages, these compounds lowered TC accumulation in HMMs during foam cell formation by limiting the uptake of acLDL and enhancing UC efflux. They may offer promise as drug therapies for atherosclerosis. PMID- 10479664 TI - Efficient nuclear delivery of antisense oligodeoxynucleotides and selective inhibition of CETP expression by apo E peptide in a human CETP-stably transfected CHO cell line. AB - N,N-Dipalmitylglycyl-apolipoprotein E (129-169) peptide (dpGapoE) is an efficient gene delivery system for both plasmids and antisense oligodeoxynucleotides (ODNs). To develop a new and efficient approach to the regulation of cholesteryl ester transfer protein (CETP) expression, we used dpGapoE to transfect phosphorothioate antisense ODNs against nucleotides 329 to 349 of human CETP cDNA into a human CETP-stably transfected Chinese hamster ovary (CHO) cell line (hCETP CHO). After transfection, translocation to the nuclei and concentration in nuclear structures were observed in >95% of the cells at 6 and 12 hours by fluorescence microscopy. No membrane disruption was observed after transfection of ODNs by dpGapoE. Although the translocation stability of phosphorothioate ODNs in the nuclei continued for >48 hours, it had weakened after 24 hours. Cellular CETP mRNA levels gradually declined, and the maximum reduction in the mRNA level (>50%) was observed at 36 hours, after which the mRNA level started to recover. CETP activity in the culture medium declined over 72 hours. The maximum reduction in CETP activity was observed at 36 hours (53.8% of control). Neither CETP mRNA nor CETP activities changed throughout the experiment after the transfection of sense phosphorothioate ODNs delivered by dpGapoE complex or naked antisense ODNs. We conclude that (1) the novel synthetic dpGapoE was a highly effective and nontoxic vehicle for the nuclear delivery of antisense ODNs into hCETP-CHO cells and (2) antisense ODNs selectively inhibited both CETP expression and activity in an hCETP-CHO cell line. This approach may enable gene regulation in vivo and could possibly be used as an antiatherosclerotic agent to alter high density lipoprotein metabolism. PMID- 10479665 TI - Human serum Paraoxonase/Arylesterase's retained hydrophobic N-terminal leader sequence associates with HDLs by binding phospholipids : apolipoprotein A-I stabilizes activity. AB - In serum, human paraoxonase/arylesterase (PON1) is found exclusively associated with high density lipoprotein (HDL) and contributes to its antiatherogenic properties by inhibiting low density lipoprotein (LDL) oxidation. Difficulties in purifying PON1 from apolipoprotein A-I (apoA-I) suggested that PON1's association with HDL may occur through a direct binding between these 2 proteins. An unusual property of PON1 is that the mature protein retains its hydrophobic N-terminal signal sequence. By expressing in vitro a mutant PON1 with a cleavable N terminus, we demonstrate that PON1 associates with lipoproteins through its N terminus by binding phospholipids directly rather than binding apoA-I. Nonetheless, apoA-I stabilized arylesterase activity more than did phospholipid alone, apoA-II, or apoE. Consequently, we studied the role of apoA-I in PON1 expression and HDL association in mice genetically deficient in apoA-I. Though present in HDL fractions at decreased levels, PON1 arylesterase activity was less stable than in control mice. Furthermore, PON1 could be competitively removed from HDL by phospholipids, suggesting that PON1's retained N-terminal peptide allows transfer of the enzyme between phospholipid surfaces. Thus, our data suggest that PON1 is stabilized by apoA-I, and its binding to HDL and physiological distribution are dependent on the direct binding of the retained hydrophobic N-terminus to phospholipids optimally presented in association with apoA-I. PMID- 10479666 TI - Decreased protection by HDL from poorly controlled type 2 diabetic subjects against LDL oxidation may Be due to the abnormal composition of HDL. AB - High plasma triglyceride concentrations in diabetic subjects increase their risk for developing coronary heart disease. Numerous studies have shown that the high density lipoprotein (HDL) composition is abnormal in type 2 diabetic subjects. One study has shown that HDL (lipoprotein A-I) isolated from subjects with non insulin-dependent diabetes mellitus exhibits a decreased capacity to induce cholesterol efflux. The current study examined the effect of HDL(2) and HDL(3) subfractions from poorly controlled type 2 diabetic and control subjects on THP-1 macrophage-mediated low density lipoprotein (LDL) oxidation. The composition and protective effects of HDL(2), but not of HDL(3), differed significantly between control and diabetic subjects. HDL(2) from diabetics were triglyceride enriched and cholesterol depleted compared with those from controls. Control HDL(2) inhibited LDL oxidation, as assessed by lipid peroxides and electrophoretic mobility, significantly (P<0.05) more than did diabetic HDL(2) in both the fasting and postprandial state. In addition, HDL(2) from diabetics did not protect against apolipoprotein B-100 fragmentation in LDL. Cross-linking in apolipoprotein A-I, oxidized in the presence of LDL, was extensive in HDL(2) from diabetics compared with that from controls. Serum triglyceride concentrations were negatively correlated with protection by HDL(2) (r=-0.673, P<0.05) in diabetic but not in control subjects. HDL(2)-associated platelet-activating factor acetylhydrolase activity was positively correlated with protection by HDL(2) in control (r=0.872, P<0.002) but not in diabetic subjects. In conclusion, compositional alterations in HDL(2) from poorly controlled type 2 diabetic subjects may reduce its antiatherogenic properties. PMID- 10479667 TI - LDL size in African Americans, Hispanics, and non-Hispanic whites : the insulin resistance atherosclerosis study. AB - The prevalence of cardiovascular disease (CVD) and atherosclerosis varies among several minority ethnic groups in the United States. Recently, small, dense low density lipoprotein (LDL) particle size has been recognized as a risk factor for CVD. We examined LDL size as a possible explanation for differences in CVD rates in 1571 subjects from the Insulin Resistance Atherosclerosis Study (IRAS), a multiethnic study of insulin resistance and cardiovascular risk factors. LDL size (A) was significantly different by ethnic group (African Americans 262.1+/-0.6, Hispanics 257.6+/-0.6, and non-Hispanic whites 259.2+/-0.4, P<0.001). Ethnic differences in LDL size continued to be statistically significant after adjustment for upper body adiposity, insulin resistance, and glucose tolerance status. However, after further adjustment for other cardiovascular risk factors, especially ethnic differences in triglyceride and high density lipoprotein (HDL) cholesterol levels, the ethnic differences in LDL size were markedly attenuated and in general no longer statistically significant. The relation of triglyceride, HDL cholesterol, insulin resistance, and adiposity to LDL size in each ethnic group was similar. LDL size differs by ethnic group, which is independent of obesity or insulin resistance. These ethnic differences appear to be due to ethnic variations in dyslipidemia (especially differences in triglyceride levels); ethnic differences in LDL size are not consistent with previously reported ethnic dissimilarities in CVD or atherosclerosis. PMID- 10479668 TI - Cystatin C as a determinant of fasting plasma total homocysteine levels in coronary artery disease patients with normal serum creatinine. AB - Serum creatinine, a surrogate for both renal function and homocysteine generation, is a determinant of fasting plasma total homocysteine levels in coronary artery disease (CAD) patients. We hypothesized that among stable-CAD patients with normal creatinine levels (ie, 0.2). Consistent with the impact of folic acid fortification of cereal grain flour in the general population, only 1 of the CAD subjects (0.6%) had a plasma folate level <3 ng/mL. We conclude that serum cystatin C levels may reflect subtle decreases in renal function that independently predict fasting total homocysteine levels among stable-CAD patients with normal serum creatinine. PMID- 10479669 TI - Kininogens are antithrombotic proteins In vivo. AB - Kininogens have recently been shown to possess antiadhesive, anticoagulant, and profibrinolytic properties and can inhibit platelet activation at low thrombin concentrations. To test whether kininogens have antithrombotic properties in vivo, we devised a model of limited arterial injury confined to removal of the endothelium. Brown-Norway Katholiek strain rats with an absence of low- and high molecular-weight kininogen due to a single point mutation, A163T, were compared in the thrombosis model to the wild-type animals, which were otherwise genetically identical. Despite an equivalent vascular injury, the mean time (+/ SEM) for a 90% decrease in flow measured by laser Doppler was 38.4+/-17 minutes in the kininogen-deficient rats compared with 194+/-29 minutes in the wild-type animals (P<0.002). The degree of vascular injury was the same. No evidence for disseminated intravascular coagulation (decrease in factor V, antithrombin, or fibrinogen) or excessive fibrinolysis (elevation of fibrinogen degradation products) was found in either group of animals. The results suggest that kininogens have antithrombotic properties at low concentrations of thrombin and that inhibitory peptides derived from kininogen may constitute a new antithrombotic strategy. PMID- 10479670 TI - Cellular effects of heparin on the production and release of tissue factor pathway inhibitor in human endothelial cells in culture. AB - Tissue factor pathway inhibitor (TFPI), the major downregulator of procoagulant activity of the tissue factor-factor VIIa complex (TF. FVIIa), is synthesized and constitutively secreted by endothelial cells (ECs). Here we describe the in vitro effects of heparin on the cellular localization, gene expression, and release of TFPI in human ECs in culture. Both unfractionated heparin (UFH) and low-molecular weight heparin (LMWH; Fragmin) time-dependently induced a significant enhanced secretion of TFPI, paralleled by a redistribution and increase of TFPI on the cell surface and a decrease of intracellular TFPI. Immunogold electron microscopy showed the presence of clusters of TFPI, both on the plasmalemma proper and within cell-surface opened caveolae/enlarged caveolar profiles. Activation of FX by TF. FVIIa on ECs treated with endotoxin was inhibited by both heparins but to a higher extent by LMWH. Inhibition of protein synthesis by cycloheximide did not reduce the release of TFPI induced by heparin. Long-term incubation (48 hours) resulted in a time-dependent enhanced production of TFPI. After the first 4 to 8 hours, depletion of intracellular TFPI was observed, more significantly with UFH. Northern blot analysis of TFPI mRNA also showed a decrease of the 1.4-kb transcript after 4 hours of incubation with UFH, followed by recovery and an increase over the control level after 24 hours. Incubation of ECs with phorbol ester (PMA) significantly enhanced the secretion of TFPI and increased its activity on the cell surface, probably by preventing invagination of caveolae. Heparin-stimulated release of TFPI decreased significantly in the presence of PMA to a level that was 2. 4 times lower than the expected additive value for PMA and UFH separately. Pretreatment of ECs with PMA suppressed a subsequent response to heparin. Altogether, our results suggest that the heparin-induced release of TFPI might involve a more specific mechanism(s) than the previously hypothesized simple displacement of TFPI from the cell surface glycocalyx. We assume that the increased secretion and redistribution of cellular TFPI induced by heparins in ECs in culture can play an important role in the modulation of the anticoagulant properties of the endothelium. PMID- 10479672 TI - Antithrombotic efficacy of the vitamin K antagonist fluindione in a human Ex vivo model of arterial thrombosis : effect of anticoagulation level and combination therapy with aspirin. AB - Thrombin is a main mediator of arterial thrombus formation, and its inhibition is an important antithrombotic strategy. However, the place of vitamin K antagonists among the different therapeutic strategies for preventing arterial thrombus formation is still debated. We studied the antithrombotic efficacy of the vitamin K antagonist fluindione in a human ex vivo model of arterial thrombosis and determined whether aspirin enhances fluindione efficacy. Ten healthy male volunteers were randomly assigned to receive fluindione, alone or in combination with aspirin (325 mg/d). Fluindione was given at increasing doses to give a stable international normalized ratio (INR) between 1.5 and 2.0 and between 2.1 and 3.0. We induced arterial thrombus formation ex vivo by exposing collagen- or tissue factor (TF)-coated coverslips in a parallel-plate perfusion chamber to native blood for 3 minutes at an arterial wall shear rate of 2600 s(-1). Platelet and fibrin deposition were measured by immunoenzymatic methods. Fluindione inhibited thrombus formation on TF-coated coverslips in a dose-dependent manner by 50% and 80% at INR 1.5 to 2.0 and INR 2.1 to 3.0, respectively (P<0.05). Fluindione in combination with aspirin inhibited TF-induced thrombus formation in a comparable manner. Collagen-induced thrombus formation was not reduced in subjects treated by fluindione. It was reduced by 50% to 60% in those treated with fluindione plus aspirin, regardless of the level of anticoagulation (P<0.05). Thus, the effectiveness of fluindione for preventing arterial thrombosis is dependent on the nature of the thrombogenic trigger. Fluindione is very effective in preventing TF- but not collagen-triggered thrombus formation. Aspirin enhances the antithrombotic effectiveness of fluindione, because combined treatment interrupts both TF- and collagen-induced thrombus formation. PMID- 10479671 TI - Tissue factor pathway inhibitor attenuates procoagulant activity and upregulation of tissue factor at the site of balloon-induced arterial injury in pigs. AB - Intravenous infusion of recombinant tissue factor pathway inhibitor (rTFPI) for 24 hours decreases neointimal thickening and luminal stenosis 1 month after balloon-induced injury to the carotid arteries in minipigs. This study was designed to determine whether the effect of rTFPI is accounted for by early decreases in procoagulant activity and thrombosis on the injured vessel wall. Vascular injury was induced by balloon hyperinflations in both carotid arteries of anesthetized pigs given no anticoagulant as a control (n=16), an intravenous infusion for 24 hours of rTFPI (0.5 mg/kg bolus and 25 microg. kg(-1). min(-1), n=14), or an intravenous infusion of unfractionated heparin (100 U. kg(-1). h( 1), n=19). Accumulation of radiolabeled autologous platelets was markedly decreased over 24 hours on injured arteries from animals given rTFPI (0.6x10(6)/cm(2)) compared with controls (2.5x10(6)/cm(2), P=0.0004). Deposition of radiolabeled fibrin was also decreased in rTFPI-treated animals (269+/-266 microg/cm(2)) compared with controls (2389+/-1673 microg/cm(2), P=0.04). Similar effects were observed with heparin. However, factor Xa activity, assayed after 24 hours by incubation of the injured arterial segments with the chromogenic substrate S-2222, was decreased more markedly on arteries from rTFPI-treated animals (0.14+/-0.13 OD) than those from heparin-treated animals (0.29+/-0.18 OD) compared with controls (0. 47+/-0.24 OD, P=0.0007). In addition, arteries from rTFPI-treated animals showed a 4-fold lower induction of tissue factor protein compared with controls (P=0.0002). Attenuation of procoagulant activity and tissue factor-mediated thrombin generation in response to injury may account for the promising results with rTFPI in the porcine angioplasty model. PMID- 10479673 TI - Endotoxin induces a second window of protection in the rat heart as determined by using p-nitro-blue tetrazolium staining, cardiac troponin T release, and histology. AB - Pretreatment of rats with small doses of lipopolysaccharide (LPS), eg, for 24 hours, attenuates the cardiac dysfunction caused by subsequent period of myocardial ischemia. This phenomenon of enhanced tolerance to an ischemic insult has been termed "second window of protection." Although the cardioprotective effects of LPS were first reported in 1989, it is still unclear whether the observed attenuation by LPS of the ischemia-induced cardiac dysfunction is indeed secondary to the protection of cardiac myocytes against ischemic cell injury and death. This study was designed to investigate the effects of "preconditioning" with LPS on cell injury caused by regional myocardial ischemia and reperfusion in the anesthetized rat. Thirty-five Wistar rats were subjected to 25 minutes occlusion of the left anterior descending coronary artery followed by 2 hours of reperfusion. Hemodynamic parameters were continuously recorded, and at the end of the experiments, infarct size (using p-nitro-blue tetrazolium staining), cardiac troponin T release, and histological markers of cell injury and death were determined. In rats pretreated with a bolus of saline (vehicle for LPS) 2 or 24 hours before left anterior descending coronary artery occlusion and reperfusion, the infarct size was 59+/-4% (2 hours saline-control, n=6) and 61+/-3% (24 hours saline-control, n=6), respectively. Pretreatment of animals with a bolus of LPS (1 mg/kg IP) 24 hours before the onset of myocardial ischemia and reperfusion reduced both infarct size (to 18+/-7%; P<0.05, n=6) as well as histological signs of cell injury. Pretreatment (24 hours, as above) of rats with LPS also reduced the release of cardiac troponin T from 58+/-13 ng/mL (saline-control) to 16+/-9 ng/mL. In contrast, pretreatment of rats with LPS (2 hours, as above) did not affect infarct size (56+/-8%, n=6), cardiac troponin T release, or the histological parameters of cell injury. These data provide the first conclusive evidence that pretreatment of rats with a bolus of LPS 24 hours before intervention reduces the cell injury and death caused by a subsequent period of myocardial ischemia and reperfusion. PMID- 10479674 TI - Membrane trafficking regulates the activity of the human dopamine transporter. AB - The trafficking of synaptic proteins is unquestionably a major determinant of the properties of synaptic transmission. Here, we present a detailed analysis of the downregulation and intracellular trafficking of the cocaine- and amphetamine sensitive dopamine transporter (DAT), a presynaptic plasma membrane protein responsible for the regulation of extracellular DA concentrations. Using PC12 cells stably transfected with human DAT cDNA, we observe that phorbol ester activation of protein kinase C (PKC) results in decreased transporter capacity and a parallel decrease in the amount of DAT on the cell surface that is attributable to intracellular transporter sequestration. After internalization, DAT diverges to the recycling, as opposed to the degradative, arm of the endocytic pathway. This study demonstrates, for the first time, DAT endocytosis, establishes the pathways through which DAT traffics both at steady state and in response to PKC activation, and suggests that DAT recycling is likely to occur. PMID- 10479675 TI - Neuronal expression of neural nitric oxide synthase (nNOS) protein is suppressed by an antisense RNA transcribed from an NOS pseudogene. AB - Here, we show that a nitric oxide synthase (NOS) pseudogene is expressed in the CNS of the snail Lymnaea stagnalis. The pseudo-NOS transcript includes a region of significant antisense homology to a previously reported neuronal NOS (nNOS) encoding mRNA. This suggested that the pseudo-NOS transcript acts as a natural antisense regulator of nNOS protein synthesis. In support of this, we show that both the nNOS-encoding and the pseudo-NOS transcripts are coexpressed in giant identified neurons (the cerebral giant cells) in the cerebral ganglion. Moreover, reverse transcription-PCR experiments on RNA isolated from the CNS establish that stable RNA-RNA duplex molecules do form between the two transcripts in vivo. Using an in vitro translation assay, we further demonstrate that the antisense region of the pseudogene transcript prevents the translation of nNOS protein from the nNOS-encoding mRNA. By analyzing NOS RNA and nNOS protein expression in two different identified neurons, we find that when both the nNOS-encoding and the pseudo-NOS transcripts are present in the same neuron, nNOS enzyme activity is substantially suppressed. Importantly, these results show that a natural antisense mechanism can mediate the translational control of nNOS expression in the Lymnaea CNS. Our findings also suggest that transcribed pseudogenes are not entirely without purpose and are a potential source of a new class of regulatory gene in the nervous system. PMID- 10479676 TI - Expression of Ca(2+)-mobilizing endothelin(A) receptors and their role in the control of Ca(2+) influx and growth hormone secretion in pituitary somatotrophs. AB - The expression and coupling of endothelin (ET) receptors were studied in rat pituitary somatotrophs. These cells exhibited periods of spontaneous action potential firing that generated high-amplitude fluctuations in cytosolic calcium concentration ([Ca(2+)](i)). The message and the specific binding sites for ET(A), but not ET(B), receptors were found in mixed pituitary cells and in highly purified somatotrophs. The activation of these receptors by ET-1 led to an increase in inositol 1,4,5-trisphosphate production and the associated rise in [Ca(2+)](i) and growth hormone (GH) secretion. The Ca(2+)-mobilizing action of ET 1 lasted for 2-3 min and was followed by an inhibition of action potential-driven Ca(2+) influx and GH secretion to below the basal levels. As in somatostatin treated cells, the ET-1-induced inhibition of spontaneous electrical activity and Ca(2+) influx was accompanied by the inhibition of adenylyl cyclase and by the stimulation of inward rectifier potassium current. In contrast to somatostatin, ET-1 did not inhibit voltage-gated Ca(2+) channels. During prolonged agonist stimulation a gradual recovery of Ca(2+) influx and GH secretion occurred. In somatotrophs treated with pertussis toxin overnight, the ET-1-induced Ca(2+) mobilizing phase was preserved, but it was followed immediately by facilitated Ca(2+) influx and GH secretion. Both somatostatin- and ET-1-induced inhibitions of adenylyl cyclase activity were abolished in pertussis toxin-treated cells. These results indicate that the transient cross-coupling of Ca(2+)-mobilizing ET(A) receptors to the G(i)/G(o) pathway in somatotrophs provides an effective mechanism to change the rhythm of [Ca(2+)](i) signaling and GH secretion during continuous agonist stimulation. PMID- 10479677 TI - Insulin prohormone processing, distribution, and relation to metabolism in Aplysia californica. AB - The first Aplysia californica insulin gene is characterized and its proteolytic processing from prohormone to final peptides elucidated using a combination of biochemical and mass spectrometric methods. Aplysia insulin (AI) is one of the largest insulins found, with a molecular weight of 9146 Da, and an extended A chain compared with other invertebrate and vertebrate insulins. The AI prohormone produces a series of C peptides and also a unique N-terminally acetylated D peptide. AI-producing cells are restricted to the central region of the cerebral ganglia mostly within the F and C clusters, and AI is transported to neurohemal release sites located on the upper labial and anterior tentacular nerves. The expression of AI mRNA decreases when the animal is deprived of food, and injections of AI reduce hemolymph glucose levels, suggesting that the function of insulin-regulating metabolism has been conserved. PMID- 10479678 TI - Two types of K(+) channel subunit, Erg1 and KCNQ2/3, contribute to the M-like current in a mammalian neuronal cell. AB - The potassium M current was originally identified in sympathetic ganglion cells, and analogous currents have been reported in some central neurons and also in some neural cell lines. It has recently been suggested that the M channel in sympathetic neurons comprises a heteromultimer of KCNQ2 and KCNQ3 (Wang et al., 1998) but it is unclear whether all other M-like currents are generated by these channels. Here we report that the M-like current previously described in NG108-15 mouse neuroblastoma x rat glioma cells has two components, "fast" and "slow", that may be differentiated kinetically and pharmacologically. We provide evidence from PCR analysis and expression studies to indicate that these two components are mediated by two distinct molecular species of K(+) channel: the fast component resembles that in sympathetic ganglia and is probably carried by KCNQ2/3 channels, whereas the slow component appears to be carried by merg1a channels. Thus, the channels generating M-like currents in different cells may be heterogeneous in molecular composition. PMID- 10479679 TI - Differential expression of brain-derived neurotrophic factor, neurotrophin-3, and neurotrophin-4/5 in the adult rat spinal cord: regulation by the glutamate receptor agonist kainic acid. AB - Previous in vitro studies indicate that select members of the neurotrophin gene family, namely brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5), contribute to survival and differentiation of spinal cord motoneurons. To investigate the potential roles of these factors in the adult spinal cord, we examined their cellular localization and regulation after systemic exposure to an excitotoxic stimulus, kainic acid (KA). Of the neurotrophins examined, NT-4/5 mRNA was most robustly expressed in the lumbosacral spinal cord of the normal adult rat, including expression by neurons throughout the gray matter, and in a subpopulation of white and gray matter glia. Both BDNF and NT-3 mRNAs were also densely expressed by alpha motoneurons of lamina IX, but were detected at lower levels elsewhere in the gray matter. NT-3 mRNA was additionally expressed by spinal cord glia, but was less widespread compared to NT-4/5. In response to systemic administration of KA, NT-4/5 and BDNF mRNAs were dramatically upregulated in a spatially and temporally restricted fashion, whereas levels of NT-3 mRNA were unchanged. These results provide strong in vivo evidence to support the idea that BDNF, NT-3, and in particular, NT-4/5, play a role in the normal function of the adult spinal cord. Furthermore, our results indicate that the actions of BDNF and NT-4/5 participate in the response of the cord to excitotoxic stimuli, and that those of NT-4/5 and NT-3 include both neurons and glia. PMID- 10479680 TI - Neuronal interleukin-16 (NIL-16): a dual function PDZ domain protein. AB - Interleukin (IL)-16 is a proinflammatory cytokine that has attracted widespread attention because of its ability to block HIV replication. We describe the identification and characterization of a large neuronal IL-16 precursor, NIL-16. The N-terminal half of NIL-16 constitutes a novel PDZ domain protein sequence, whereas the C terminus is identical with splenocyte-derived mouse pro-IL-16. IL 16 has been characterized only in the immune system, and the identification of NIL-16 marks a previously unsuspected connection between the immune and the nervous systems. NIL-16 is a cytosolic protein that is detected only in neurons of the cerebellum and the hippocampus. The N-terminal portion of NIL-16 interacts selectively with a variety of neuronal ion channels, which is similar to the function of many other PDZ domain proteins that serve as intracellular scaffolding proteins. Among the NIL-16-interacting proteins is the class C alpha1 subunit of a mouse brain calcium channel (mbC alpha1). The C terminus of NIL-16 can be processed by caspase-3, resulting in the release of secreted IL-16. Furthermore, in cultured cerebellar granule neurons undergoing apoptosis, NIL-16 proteolysis parallels caspase-3 activation. Cerebellar granule neurons express the IL-16 receptor CD4. Exposure of these cells to IL-16 induces expression of the immediate-early gene, c-fos, via a signaling pathway that involves tyrosine phosphorylation. This suggests that IL-16 provides an autocrine function in the brain. Therefore, we hypothesize that NIL-16 is a dual function protein in the nervous system that serves as a secreted signaling molecule as well as a scaffolding protein. PMID- 10479681 TI - Alternative splicing of the C-terminal domain regulates cell surface expression of the NMDA receptor NR1 subunit. AB - Subcellular localization of the NMDA receptor NR1 splice forms was studied by expressing individual splice variants and their epitope-tagged derivatives in mouse fibroblasts and in hippocampal neurons. When NR1 splice variants were expressed in fibroblasts, the amount of NR1 molecules expressed on the cell surface varied among forms with different C-terminal cytoplasmic domains. The splice forms with the longest C-terminal cytoplasmic tail (NR1-1a and NR1-1b) showed the lowest amount of cell surface expression, and the splice forms with the shortest C-terminal cytoplasmic tail (NR1-4a and NR1-4b) showed the highest cell surface expression. Cell surface expression of NR1 was enhanced by the coexpression of the NR2 subunit. We measured the glutamate-induced increase of calcium concentration in fibroblasts expressing one of the NR1 splice forms and the NR2B subunit. The increase of calcium concentration after glutamate application had a positive correlation with the amount of NR1 splice forms expressed on the cell surface. When epitope-tagged NR1 splice variants were expressed in primary hippocampal neurons using recombinant adenoviruses, we also observed the differential expression on the cell surface between splice variants. These results suggest that the splicing of the C-terminal domain of the NR1 subunit regulates the cell surface expression of the functional NMDA receptors. PMID- 10479682 TI - The Drosophila beta-amyloid precursor protein homolog promotes synapse differentiation at the neuromuscular junction. AB - Although abnormal processing of beta-amyloid precursor protein (APP) has been implicated in the pathogenic cascade leading to Alzheimer's disease, the normal function of this protein is poorly understood. To gain insight into APP function, we used a molecular-genetic approach to manipulate the structure and levels of the Drosophila APP homolog APPL. Wild-type and mutant forms of APPL were expressed in motoneurons to determine the effect of APPL at the neuromuscular junction (NMJ). We show that APPL was transported to motor axons and that its overexpression caused a dramatic increase in synaptic bouton number and changes in synapse structure. In an Appl null mutant, a decrease in the number of boutons was found. Examination of NMJs in larvae overexpressing APPL revealed that the extra boutons had normal synaptic components and thus were likely to form functional synaptic contacts. Deletion analysis demonstrated that APPL sequences responsible for synaptic alteration reside in the cytoplasmic domain, at the internalization sequence GYENPTY and a putative G(o)-protein binding site. To determine the likely mechanisms underlying APPL-dependent synapse formation, hyperexcitable mutants, which also alter synaptic growth at the NMJ, were examined. These mutants with elevated neuronal activity changed the distribution of APPL at synapses and partially suppressed APPL-dependent synapse formation. We propose a model by which APPL, in conjunction with activity-dependent mechanisms, regulates synaptic structure and number. PMID- 10479683 TI - Fast excitatory nicotinic transmission in the chick lateral spiriform nucleus. AB - The lateral spiriform nucleus (SpL) in the chick mesencephalon contains functional nicotinic receptors and receives a cholinergic fiber projection. We now use double-label immunohistochemistry to demonstrate that choline acetyltransferase-immunopositive fibers in the SpL and in the cholinergic fiber tract lateral to the nucleus are associated with fibers expressing the alpha5 and/or alpha3 nicotinic receptor subunits as determined by mAb35 immunoreactivity. This morphological evidence suggests that there might be synapses between the cholinergic fibers and the dendrites of SpL neurons. Whole cell recordings from SpL neurons in current-clamp mode revealed EPSPs evoked by stimulation of the cholinergic fiber tract lateral to the SpL. These EPSPs increased in amplitude in the presence of bicuculline. Further addition of the nicotinic antagonist dihydro-beta-erythroidine (DHbetaE) to the buffer significantly attenuated them. Almost all of the remaining EPSP was blocked by 6,7-dinitroquinoxaline-2,3-dione. In the presence of an antagonist cocktail that isolated the nicotinic responses, a fast, monosynaptic nicotinic EPSP or EPSC was evoked. In some neurons, the nicotinic EPSP resulted in the generation of an action potential. The nicotinic nature of the evoked response was confirmed by blockade of the EPSPs or EPSCs with nicotinic antagonists, including DHbetaE, D tubocurare, and mecamylamine. The nicotinic response was insensitive to low concentrations (10-100 nM) of methyllycaconitine, indicating that typical alpha7 containing receptors were not involved. The results demonstrate that endogenously released acetylcholine generates EPSPs that can elicit action potentials by acting at postsynaptic nicotinic receptors on SpL neurons. PMID- 10479684 TI - Composition of the GABA(A) receptors of retinal dopaminergic neurons. AB - Transgenic technology, single-cell RT-PCR, and immunocytochemistry were combined to investigate the composition of the GABA(A) receptors of dopaminergic (interplexiform) amacrine (DA) cells. A mouse line was used in which these neurons were labeled with human placental alkaline phosphatase and could therefore be identified in vitro after dissociation of the retina. We performed single-cell RT-PCR on the isolated cells and showed that (1) DA cells contained the messages for alpha1, alpha3, alpha4, beta1, beta3, gamma1, gamma2(S), and gamma2(L) subunits; (2) this transcript repertory did not change on dissociation of the retina and throughout the time required for cell harvesting; and (3) all DA cells contained the entire transcript repertory. Immunocytochemistry with subunit-specific antibodies showed that all subunits were expressed and appeared homogeneously distributed throughout the cell membrane at a low concentration. In addition, with the exception of alpha4, the subunits formed clusters at the surface of the dendrites and on the inner pole of the cell body. Because of their size, shape, and topographic coincidence with GABAergic endings, the clusters were interpreted as postsynaptic active zones containing GABA(A) receptors. The composition of the synaptic receptors was not uniform: clusters distributed throughout the dendritic tree contained alpha3, beta3, and, less frequently, beta1 subunits, whereas clusters containing the alpha1 subunit were confined to large dendrites. Therefore, DA cells possess at least two types of GABA(A) receptors localized in different synapses. Furthermore, they exhibit multiple extrasynaptic GABA(A) receptors. PMID- 10479685 TI - Tetanic stimulation leads to increased accumulation of Ca(2+)/calmodulin dependent protein kinase II via dendritic protein synthesis in hippocampal neurons. AB - mRNA for the alpha-subunit of CaMKII is abundant in dendrites of neurons in the forebrain (Steward, 1997). Here we show that tetanic stimulation of the Schaffer collateral pathway causes an increase in the concentration of alpha-CaMKII in the dendrites of postsynaptic neurons. The increase is blocked by anisomycin and is detected by both quantitative immunoblot and semiquantitative immunocytochemistry. The increase in dendritic alpha-CaMKII can be measured 100 200 micrometer away from the neuronal cell bodies as early as 5 min after a tetanus. Transport mechanisms for macromolecules from neuronal cell bodies are not fast enough to account for this rapid increase in distal portions of the dendrites. Therefore, we conclude that dendritic protein synthesis must produce a portion of the newly accumulated CaMKII. The increase in concentration of dendritic CaMKII after tetanus, together with the previously demonstrated increase in autophosphorylated CaMKII (Ouyang et al., 1997), will produce a prolonged increase in steady-state kinase activity in the dendrites, potentially influencing mechanisms of synaptic plasticity that are controlled through phosphorylation by CaMKII. PMID- 10479686 TI - Lamina-specific synaptic activation causes domain-specific alterations in dendritic immunostaining for MAP2 and CAM kinase II. AB - Polyribosomal complexes are selectively localized beneath postsynaptic sites on neuronal dendrites; this localization suggests that the translation of the mRNAs that are present in dendrites may be regulated by synaptic activity. The present study tests this hypothesis by evaluating whether synaptic activation alters the immunostaining pattern for two proteins whose mRNAs are present in dendrites: the dendrite-specific cytoskeletal protein MAP2 and the alpha-subunit of CAMKII. High frequency stimulation of the perforant path projections to the dentate gyrus, which terminate in a discrete band on the dendrites of dentate granule cells, produced a two-stage alteration in immunostaining for MAP2 in the dendritic laminae. Five minutes of stimulation (30 trains) caused a decrease in MAP2 immunostaining in the lamina in which the activated synapses terminate. After more prolonged periods of stimulation (1-2 hr), there was an increase in immunostaining in the sideband laminae just proximal and distal to the activated band of synapses. The same stimulation paradigm produced a modest increase in immunostaining for alpha-CAMKII in the activated laminae, with no detectable changes in the sideband laminae. The alterations in immunostaining for MAP2 were diminished, but not eliminated, by inhibiting protein synthesis; the increases in CAMKII were not. These findings reveal that patterned synaptic activity can produce domain-specific alterations in the molecular composition of dendrites; these alterations may be caused in part by local protein synthesis and in part by other mechanisms. PMID- 10479687 TI - Measurement of action potential-induced presynaptic calcium domains at a cultured neuromuscular junction. AB - Spatially localized Ca(2+) domains are thought to play a key role in action potential (AP)-evoked neurotransmitter release at fast synapses. We used a stage scan confocal spot-detection method and the low-affinity Ca(2+) indicator Oregon Green 488 BAPTA-5N to study the spatiotemporal profile of presynaptic AP-induced Ca(2+) domains. Families of scanned AP-induced fluorescence transients were detected from spot locations separated by 200-300 nm, within the vicinity of Ca(2+) entry sites. Typically, the largest transient in a particular scan peaked within approximately 1 msec and decayed with rapid (tau(1) of 1.7 msec) and slow components (tau(2) of 16 msec, tau(3) of 78 msec). As the spot was incrementally displaced, transients progressively exhibited a slowing in their time-to-peak and a loss of the fast decay component. Three-dimensional graphs of fluorescence versus time and spot displacement revealed the presence of AP-induced fluorescence domains that dissipated within approximately 7 msec. The size of fluorescence domains were estimated from the full-width at half-maximum of gaussian fits to isochronal DeltaF/F plots and ranged from 0.6 to 3.0 micrometer, with a mean +/- SD of 1.6 +/- 0.6 micrometer. Model simulations of a localized Ca(2+) entry site predicted the major features of the fluorescence transients and suggested that, within approximately 1 msec of the initiation of the Ca(2+) current, both the fluorescence domain and the underlying Ca(2+) domain do not extend significantly beyond the site of entry. Consistent with this prediction, the intracellular addition of EGTA (up to 2 mM) accelerated the decay of the measured transients but did not affect the domain size. PMID- 10479688 TI - Bax-dependent caspase-3 activation is a key determinant in p53-induced apoptosis in neurons. AB - p53 is a pivotal molecule regulating the death of neurons both after acute injury and during development. The molecular mechanisms by which p53 induces apoptosis in neuronal cells, however, are not well understood. We have shown previously that adenovirus-mediated p53 gene delivery to neurons was sufficient to induce apoptosis. In the present study we have examined the molecular mechanism by which p53 evokes neuronal cell death. Adenovirus-mediated delivery of p53 to cerebellar granule neurons resulted in caspase-3 (CPP32) activation followed by terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) staining and loss of viability as determined by an MTT survival assay. To determine whether Bax is essential for caspase-3 activation, p53 was expressed in Bax-deficient cells. Bax null neurons did not exhibit caspase-3 activation in response to p53 and were protected from apoptosis. To determine whether Bax dependent caspase-3 activation was required in p53-mediated neuronal cell death, caspase-3-deficient neurons were examined. Our results indicate that caspase-3 deficient neurons exhibit a remarkable delay in apoptosis and a dramatic decrease in TUNEL-positive cells. These studies demonstrate that p53-induced cell death in postmitotic neurons involves a Bax-dependent caspase-3 activation, suggesting that these molecules are important determinants in neuronal cell death after injury. PMID- 10479689 TI - A dominant negative receptor for specific secreted semaphorins is generated by deleting an extracellular domain from neuropilin-1. AB - Neuropilins have recently been characterized as receptors for secreted semaphorins. Here, we report the generation of a dominant negative form of neuropilin-1 by the deletion of one of its extracellular domains. Expression of this variant in cultured primary sympathetic neurons blocks the paralysis of growth cone motility normally induced by SEMA-3A (collapsin-1, semaphorin III, semaphorin D) and SEMA-3C (collapsin-3, semaphorin E) but not that induced by SEMA-3F (semaphorin IV). A truncated form of neuropilin-1 that is missing its cytoplasmic domain fails to act as a dominant negative receptor component. These results suggest that neuropilin-1 is a necessary component of receptor complexes for some, but not all, secreted semaphorin family members. Overexpression of dominant negative neuropilins should provide a powerful new method of blocking the functions of secreted semaphorins. PMID- 10479690 TI - The medial ganglionic eminence gives rise to a population of early neurons in the developing cerebral cortex. AB - During development of the neocortex, the marginal zone (layer I) and the subplate (layer VII) are the first layers to form from a primordial plexiform neoropil. The cortical plate (layers II-VI) is subsequently established between these superficial and deep components of the primordial plexiform neuropil. Neurons in the early zones are thought to play important roles in the formation of the cortex: the Cajal-Retzius cells of the marginal zone are instrumental in neuronal migration and laminar formation, and cells of the subplate are involved in the formation of cortical connections. Using the fluorescent tracer 1,1'-dioctodecyl 3,3,3', 3'-tetramethylindocarbocyanine (DiI), we have shown here that a substantial proportion of neurons of the marginal zone, including cells with features of Cajal-Retzius cells, and of the subplate and lower intermediate zone are not born in the ventricular neuroepithelium but instead originate in the medial ganglionic eminence (MGE), the pallidal primordium. These neurons follow a tangential migratory route to their positions in the developing cortex. They express the neurotransmitter GABA but seem to lack the calcium binding protein calretinin; some migrating cells found in the marginal zone express reelin. In addition, migrating cells express the LIM-homeobox gene Lhx6, a characteristic marker of the MGE. It is suggested that this gene uniquely or in combination with other transcription factors may be involved in the decision of MGE cells to differentiate in situ or migrate to the neocortex. PMID- 10479691 TI - Rapid nuclear responses to target-derived neurotrophins require retrograde transport of ligand-receptor complex. AB - Target-derived neurotrophins initiate signals that begin at nerve terminals and cross long distances to reach the cell bodies and regulate gene expression. Neurotrophin receptors, Trks, themselves serve as retrograde signal carriers. However, it is not yet known whether the retrograde propagation of Trk activation reflects movement of Trk receptors from neurites to cell bodies or reflects serial activation of stationary Trk molecules. Here, we show that neurotrophins selectively applied to distal neurites of sensory neurons rapidly induce phosphorylation of the transcription factor cAMP response element-binding protein (CREB) and also cause a slower increase in Fos protein expression. Both nuclear responses require activation of neurotrophin receptors (Trks) at distal nerve endings and retrograde propagation of Trk activation to the nerve cell bodies. Using photobleach and recovery techniques to follow biologically active, green fluorescent protein (GFP)-tagged BDNF receptors (TrkB-GFP) in live cells during retrograde signaling, we show that TrkB-GFP moves rapidly from neurites to the cell bodies. This rapid movement requires ligand binding, Trk kinase activity, and intact axonal microtubules. When they reach the cell bodies, the activated TrkB receptors are in a complex with ligand. Thus, the retrograde propagation of activated TrkB from neurites to cell bodies, although rapid, reflects microtubule dependent transport of phosphorylated Trk-ligand complexes. Moreover, the relocation of activated Trk receptors from nerve endings to cell bodies is required for nuclear signaling responses. Together, these data support a model of retrograde signaling whereby rapid vesicular transport of ligand-receptor complex from the neurites to the cell bodies mediates the nuclear responses. PMID- 10479692 TI - Effects of roundabout on growth cone dynamics, filopodial length, and growth cone morphology at the midline and throughout the neuropile. AB - roundabout (robo) encodes an axon guidance receptor that controls midline crossing in the Drosophila CNS. In robo mutants, axons that normally project ipsilaterally can cross and recross the midline. Growth cones expressing Robo are believed to be repelled from the midline by the interaction of Robo and its ligand Slit, an extracellular protein expressed by the midline glia. To help understand the cellular basis for the midline repulsion mediated by Robo, we used time-lapse observations to compare the growth cone behavior of the ipsilaterally projecting motorneuron RP2 in robo and wild-type embyros. In wild-type embryos, filopodia can project across the midline but are quickly retracted. In robo mutants, medial filopodia can remain extended for longer periods and can develop into contralateral branches. In many cases RP2 produces both ipsilateral and contralateral branches, both of which can extend into the periphery. The growth cone also exhibits longer filopodia and more extensive branching both at the midline and throughout the neuropile. Cell injections in fixed stage 13 embryos confirmed and quantified these results for both RP2 and the interneuron pCC. The results suggest that Robo both repels growth cones at the midline and inhibits branching throughout the neuropile by promoting filopodial retraction. PMID- 10479693 TI - Negative regulation of oligodendrocyte differentiation by galactosphingolipids. AB - Galactocerebroside and sulfatide, major galactosphingolipid components of oligodendrocyte plasma membranes and myelin, are first expressed at a critical point, when progenitors cease to proliferate and commence terminal differentiation. We showed previously that an antibody to galactocerebroside/sulfatide arrested terminal differentiation, suggesting a role for these galactolipids in oligodendrocyte differentiation. We have now investigated the differentiation of oligodendrocytes (1) in response to other anti-galactolipid antibodies, showing that anti-sulfatide O4 but not anti galactocerebroside O1 blocks terminal differentiation, perhaps by mimicking an endogenous ligand, and (2) in a transgenic mouse unable to synthesize these lipids because of mutation of the gene for ceramide galactosyltransferase, a key enzyme for galactosphingolipid synthesis. We find that galactosyltransferase mRNA expression begins at the late progenitor [pro-oligodendroblast (Pro-OL)] stage of the lineage and that the late progenitor marker pro-oligodendroblast antigen is not synthesized in the absence of galactosyltransferase. The principal outcome of the elimination of these galactolipids is a two- to threefold enhancement in the number of terminally differentiated oligodendrocytes both in culture and in vivo. Because the general pattern of differentiation and the level of progenitor proliferation and survival appear to be unaltered in the mutant cultures, we conclude that the increased number of oligodendrocytes is caused by an increased rate and probability of differentiation. In agreement with these two experimental approaches, we present a model in which galactosphingolipids (in particular galactocerebroside and/or sulfatide) act as sensors and/or transmitters of environmental information, interacting with endogenous ligands to function as negative regulators of oligodendrocyte differentiation, monitoring the timely progress of Pro-OLs into terminally differentiating, myelin-producing oligodendrocytes. PMID- 10479694 TI - Neuromuscular activity blockade induced by muscimol and d-tubocurarine differentially affects the survival of embryonic chick motoneurons. AB - To understand better how spontaneous motoneuron activity and intramuscular nerve branching influence motoneuron survival, we chronically treated chicken embryos in ovo with either d-tubocurarine (dTC) or muscimol during the naturally occurring cell death period, assessing their effects on activity by in ovo motility measurement and muscle nerve recordings from isolated spinal cord preparations. Because muscimol, a GABA(A) agonist, blocked both spontaneous motoneuron bursting and that elicited by descending input but did not rescue motoneurons, we conclude that spontaneous bursting activity is not required for the process of normal motoneuron cell death. dTC, which rescues motoneurons and blocks neuromuscular transmission, blocked neither spontaneous nor descending input-elicited bursting and early in the cell death period actually increased burst amplitude. These changes in motoneuron activation could alter the uptake of trophic molecules or gene transcription via altered patterns of [Ca(2+)](i), which in turn could affect motoneuron survival directly or indirectly by altering intramuscular nerve branching. A good correlation was found between nerve branching and motoneuron survival under various experimental conditions: (1) dTC, but not muscimol, greatly increased branching; (2) the removal of PSA from NCAM partially reversed the effects of dTC on both branching and survival, indicating that branching is a critical variable influencing motoneuron survival; (3) muscimol, applied with dTC, prevented the effect of dTC on survival and motoneuron bursting and, to a large extent, its effect on branching. However, the central effects of dTC also appear to be important, because muscimol, which prevented motoneuron activity in the presence of dTC, also prevented the dTC induced rescue of motoneurons. PMID- 10479695 TI - Cross-modal reorganization of horizontal connectivity in auditory cortex without altering thalamocortical projections. AB - The development of the different, highly specialized regions of the mammalian cerebral cortex depends in part on neural activity, either intrinsic spontaneous activity or externally driven sensory activity. To determine whether patterned sensory activity instructs the development of intrinsic cortical circuitry, we have experimentally altered the modality of sensory inputs to cerebral cortex. Neonatal diversion of retinal axons to the auditory thalamus (cross-modal rewiring) results in a primary auditory cortex (AI) that resembles visual cortex in its response properties and topography (Roe et al., 1990, 1992). To test the hypothesis that the visual response properties are created by a visually driven reorganization of auditory cortical circuitry, we investigated the effect of early visual experience on the development of intrinsic, horizontal connections within AI. Horizontal connections are likely to play an important role in the construction of visual response properties in AI as they do in visual cortex. Here we show that early visual inputs to auditory thalamus can reorganize horizontal connections in AI, causing both an increase in their extent and a change in pattern, so that projections are not restricted to the isofrequency axis, but extend in a more isotropic pattern around the injection site. Thus, changing afferent modality, without altering the source of the thalamocortical axons, can profoundly alter cortical circuitry. Similar changes may underlie cortical compensatory processes in deaf or blind humans and may also have played a role in the parcellation of neocortex during mammalian evolution. PMID- 10479696 TI - A neurotrophic model of the development of the retinogeniculocortical pathway induced by spontaneous retinal waves. AB - The development of the retinogeniculate pathway or the geniculocortical pathway, or both, occurs either before birth or before eye opening in many species. It is widely believed that spontaneous retinal activity could drive the segregation of afferents into eye-specific laminae or columns and the refinement of initially diffuse receptive fields and the emergence of orderly, retinotopic organization. We show that a recent computational model that generates a phenomenologically accurate representation of spontaneous retinal activity can indeed drive afferent segregation and, more particularly, topographic and receptive field refinement in the retinogeniculocortical system. We use a model of anatomical synaptic plasticity based on recent data suggesting that afferents might compete for limited amounts of retrograde neurotrophic factors (NTFs). We find that afferent segregation and receptive field formation are disrupted in the presence of exogenous NTFs. We thus predict that infusion of NTFs into the lateral geniculate nucleus should disrupt normal development and that the infusion of such factors into the striate cortex should disrupt receptive field refinement in addition to the well known disruption of ocular dominance column (ODC) formation. To demonstrate that the capacity of our model of plasticity to drive normal development is not restricted just to spontaneous retinal activity, we also use a coarse representation of visually evoked activity in some simulations. We find that such simulations can exhibit the formation of ODCs followed by their disappearance, reminiscent of the New World marmoset. A decrease in interocular correlations stabilizes these ODCs. Thus we predict that divergent strabismus should render marmoset ODCs stable into adulthood. PMID- 10479697 TI - Calcium influx alters actin bundle dynamics and retrograde flow in Helisoma growth cones. AB - The ability of calcium (Ca(2+)) to effect changes in growth cone motility requires remodeling of the actin cytoskeleton. To understand the mechanisms involved, we evaluated the effect of elevated intracellular calcium ([Ca(2+)](i)) on actin bundle dynamics, organization, and retrograde flow in the large growth cones of identified Helisoma neurons. Depolarization with 15 mM KCl (high K(+)) for 30 min caused a rapid and sustained increase in [Ca(2+)](i) and resulted in longer filopodia, shorter actin ribs, and a decrease in lamellipodia width. Time lapse microscopy revealed that increasing [Ca(2+)](i) affected actin bundle dynamics differently at the proximal and distal ends. Filopodial lengthening resulted from assembly-driven elongation of actin bundles whereas actin rib shortening resulted from a distal shift in the location of breakage. Buckling of ribs occurred before breakage, suggesting nonuniform forces were applied to ribs before shortening. Calcium (Ca(2+)) influx also resulted in a decrease in density of F-actin in bundles, as determined by contrast changes in ribs imaged by differential interference contrast microscopy and fluorescent intensity changes in rhodamine-labeled ribs. The velocity of retrograde flow decreased by 50% after elevation of [Ca(2+)](i). However, no significant change in retrograde flow occurred when the majority of changes in actin bundles were blocked by phalloidin. This suggests that inhibition of retrograde flow resulted from Ca(2+) induced changes in the actin cytoskeleton. These results implicate Ca(2+) as a regulator of actin dynamics and, as such, provide a mechanism by which Ca(2+) can influence growth cone motility and behavior. PMID- 10479698 TI - Relative contribution of endogenous neurotrophins in hippocampal long-term potentiation. AB - Recent evidence has shown that brain-derived neurotrophic factor (BDNF) is involved in hippocampal long-term potentiation (LTP). Because the reagents used in acute experiments react not only with BDNF but also with neurotrophin-4/5 (NT4/5) and neurotrophin-3 (NT3), we examined the involvement of these neurotrophins in LTP using two highly specific, function-blocking monoclonal antibodies against BDNF and NT3, as well as a TrkB-IgG fusion protein. Our results show that NT3 antibodies did not have any effects on LTP. However, both TrkB-IgG fusion proteins and BDNF antibody similarly reduced LTP, suggesting that only BDNF but no other ligands of the TrkB-receptor are likely to be involved in LTP induction. The reduction in LTP depended on the inducing stimuli and was only observed with theta-burst stimulation (TBS) but not with tetanic stimulation. We further observed that LTP was only reduced if BDNF was blocked before and during TBS stimulation, and BDNF antibodies did not affect early or late stages of LTP if they were applied 10, 30, or 60 min after TBS stimulation. These results point toward a specific and unique role of endogenous BDNF but not of other neurotrophins in the process of TBS-induced hippocampal LTP. Additionally, they suggest that endogenous BDNF is required for a limited time period only shortly before or around LTP induction but not during the whole process of LTP. PMID- 10479699 TI - Rescue of cerebellar granule cells from death in weaver NR1 double mutants. AB - The weaver mutation results in the extensive death of midline cerebellar granule cells. The mutation consists of a single base pair substitution of the gene encoding the G-protein-activated inwardly rectifying potassium channel protein, GIRK2. The functional consequences of this mutation are still in dispute. In this study we demonstrate the in vivo and in vitro rescue of weaver granule cells when NR1 NMDA subunits are eliminated in weaver NR1 double mutants. This rescue of weaver granule cells provides evidence that wvGIRK2 alone is not sufficient to cause granule cell death. PMID- 10479700 TI - Dynamic regulation of cpg15 during activity-dependent synaptic development in the mammalian visual system. AB - During visual system development, neural activity regulates structural changes in connectivity including axonal branching and dendritic growth. Here we have examined a role for the candidate plasticity gene 15 (cpg15), which encodes an activity-regulated molecule that can promote dendritic growth, in this process. We report that cpg15 is expressed in the cat visual system at relatively high levels in the lateral geniculate nucleus (LGN) but at very low levels in its synaptic target, layer 4 of the visual cortex. Prenatally, when cpg15 mRNA in the LGN is most abundant, expression is insensitive to action potential blockade by tetrodotoxin. Postnatally, activity regulation of cpg15 emerges in the LGN coincident with development of ocular dominance columns in the visual cortex. cpg15 can be detected in layers 2/3 and 5/6 of visual cortex postnatally, and expression in layers 2/3 is activity-regulated during known periods of activity dependent plasticity for these layers. Localization and regulation of cpg15 expression in the visual system are consistent with a presynaptic role for CPG15 in shaping dendritic arbors of target neurons during activity-dependent synaptic rearrangements, both in development and adulthood. PMID- 10479701 TI - Ultrastructural localization of full-length trkB immunoreactivity in rat hippocampus suggests multiple roles in modulating activity-dependent synaptic plasticity. AB - Neurotrophins acting at the trkB receptor have been shown to be important modulators of activity-dependent plasticity in the hippocampus, but the mechanisms underlying these effects are not yet well understood. To identify the cellular and subcellular targets of trkB ligands in the adult rat hippocampal formation, full-length trkB receptor immunoreactivity (trkB-IR) was localized using electron microscopy. trkB-IR was present in the glutamatergic pyramidal and granule cells. Labeling in these neurons appeared as discrete clusters and was primarily in axons, excitatory-type axon terminals, and dendritic spines and to a lesser extent in somata and dendritic shafts. trkB-IR was commonly found on the plasma membrane of dendritic spines, whereas in other subcellular regions trkB-IR was often intracellular. Labeling was strikingly dense within axon initial segments, suggesting extensive receptor trafficking. trkB-IR was not confined to pyramidal and granule cells. Dense trkB-IR was found in occasional interneuron axon initial segments, some axon terminals forming inhibitory-type synapses onto somata and dendritic shafts, and excitatory-type terminals likely to originate extrahippocampally. This suggests that trkB is contained in some GABAergic interneurons, neuromodulatory (e.g., cholinergic, dopaminergic, and noradrenergic) afferents, and/or glutamatergic afferents. These data indicate that full-length trkB receptor activation may modulate glutamatergic pathways of the trisynaptic circuit both presynaptically at axon terminals and initial segments and postsynaptically at dendritic spines and shafts. Signaling via catalytic trkB may also presynaptically affect inhibitory and modulatory neurons. A pan-trkB antibody labeled the same neuronal populations as the full-length specific trkB antiserum, but the labels differed in density at various subcellular sites. These findings provide an ultrastructural foundation for further examining the mechanisms through which neurotrophins acting at trkB receptors contribute to synaptic plasticity. PMID- 10479702 TI - Maternal aggression is reduced in neuronal nitric oxide synthase-deficient mice. AB - Lactating females express rapid extremes in behavior, ranging from gentle nurturance toward offspring to fiercely protective aggression against intruders. Although males often behave aggressively against intruders, female rodents usually express aggression only when rearing and protecting pups. Nitric oxide (NO) inhibits male aggression; however, its role in maternal aggression is unknown. In the present study, female mice with targeted disruption of the neuronal nitric oxide synthase gene (nNOS-/-) displayed significant deficits in maternal aggression relative to wild-type (WT) mice in terms of percentage displaying aggression, the average number of attacks against a male intruder, and the total time spent attacking the male intruder. The nNOS-/- mice displayed normal pup retrieval behavior. Because the specific deficits in maternal aggression in the nNOS-/- mice suggested a possible role for NO in maternal aggression, we combined behavioral testing of WT mice with immunohistochemistry for citrulline, an indirect marker of NO synthesis, to examine indirectly NO synthesis during maternal aggression. A significant increase in the number of citrulline-positive cells was identified in the medial preoptic nucleus, the suprachiasmatic nucleus, and the subparaventricular zone regions of the hypothalamus in aggressive lactating females relative to control mice. In other regions of the brain, no changes in the number of citrulline-positive cells were observed across either groups or treatments. These results provide two indirect lines of evidence that NO release is associated with maternal aggression. PMID- 10479703 TI - Reconstruction of natural scenes from ensemble responses in the lateral geniculate nucleus. AB - A major challenge in studying sensory processing is to understand the meaning of the neural messages encoded in the spiking activity of neurons. From the recorded responses in a sensory circuit, what information can we extract about the outside world? Here we used a linear decoding technique to reconstruct spatiotemporal visual inputs from ensemble responses in the lateral geniculate nucleus (LGN) of the cat. From the activity of 177 cells, we have reconstructed natural scenes with recognizable moving objects. The quality of reconstruction depends on the number of cells. For each point in space, the quality of reconstruction begins to saturate at six to eight pairs of on and off cells, approaching the estimated coverage factor in the LGN of the cat. Thus, complex visual inputs can be reconstructed with a simple decoding algorithm, and these analyses provide a basis for understanding ensemble coding in the early visual pathway. PMID- 10479704 TI - A blueprint for movement: functional and anatomical representations in the human motor system. AB - Despite a clear somatotopic organization of the motor cortex, a movement can be learned with one extremity and performed with another. This suggests that there exists a limb-independent coding for movements. To dissociate brain regions coding for movement parameters from those relevant to the chosen effector, subjects wrote their signature with their dominant index finger and ipsilateral big toe, and we determined those areas activated by both conditions using functional magnetic resonance imaging. The results show that movement parameters for this highly trained movement are stored in secondary sensorimotor cortices of the extremity with which it is usually performed, i.e., the dominant hand, including dorsal and ventral lateral premotor cortices. These areas can be accessed by the foot and are therefore functionally independent from the primary representation of the effector. Thus, somatotopy in secondary structures in the human motor system seems to be defined functionally, and not on the basis of anatomical representations. PMID- 10479705 TI - The nondiscriminating zone of directionally selective retinal ganglion cells: comparison with dendritic structure and implications for mechanism. AB - We have studied, at high resolution, the sizes and pattern of dendrites of directionally selective retinal ganglion cells in the rabbit. The dendrites had a distinctive pattern of branching. The major dendritic trunks were relatively thick, beginning at approximately 1 micrometer and tapering to approximately 0.5 micrometer in diameter. Higher order dendrites exiting from them generally stepped abruptly to a diameter of 0.4-0.6 micrometer, which they maintained throughout their length. Recording confirmed the existence of a zone within the receptive field, usually occupying 20-25% of its area, where direction of movement was only weakly discriminated. The dendritic arbors of cells, injected with Lucifer yellow after recording, revealed no difference in dendritic structure between the discriminating and nondiscriminating zones. The nondiscriminating zone was located on the preferred side of the receptive field (the side from which movement in the preferred direction originates). This is consistent with a mechanism of direction selectivity based on inhibition generated by movement in the null direction but not with feedforward excitation, as occurs in flies and is postulated in some models of mammalian direction selectivity. PMID- 10479706 TI - Effects of nonuniform fiber sensitivity, innervation geometry, and noise on information relayed by a population of slowly adapting type I primary afferents from the fingerpad. AB - The capacity of a population of primary afferent fibers to signal information about a sphere indenting the fingerpad is limited by factors such as the inhomogeneity of sensitivity among the afferents, the pattern and density of innervation, and the effects of noise (response variability). Using experimental data recorded from single slowly adapting type I afferents (SAIs), we simulated the response of the SAI population to such a stimulus. The human ability to discriminate stimulus curvature, location, and force has been quantified previously. We devised three neural measures, treating them as surrogates for the real neural measures underlying human performance, and explored how population parameters usually overlooked in neural coding studies affect such measures. Variation in sensitivity among SAIs is large; this distorts population response profiles markedly but has no significant impact on the neural measures. Two classes of noise were introduced, one dependent on and the other independent of the level of neural activity. Resolution of the model was compared with discrimination in humans. Correlation of noise among neurons had different effects for the different measures. An increase in correlation decreased resolution in the measure for force but improved resolution in the measure for position. Increasing innervation density (1) always increased resolution for position and (2) increased resolution for force if noise was uncorrelated but had diminishing effects as correlation increased. Correlation and innervation density had complex effects on the measure for curvature, depending on the class of noise. Nonuniformity in the pattern of innervation had negligible effects on resolution. PMID- 10479707 TI - Serotonin differentially modulates responses to tones and frequency-modulated sweeps in the inferior colliculus. AB - Although almost all auditory brainstem nuclei receive serotonergic innervation, little is known about its effects on auditory neurons. We address this question by evaluating the effects of serotonin on sound-evoked activity of neurons in the inferior colliculus (IC) of Mexican free-tailed bats. Two types of auditory stimuli were used: tone bursts at the neuron's best frequency and frequency modulated (FM) sweeps with a variety of spectral and temporal structures. There were two main findings. First, serotonin changed tone-evoked responses in 66% of the IC neurons sampled. Second, the influence of serotonin often depended on the type of signal presented. Although serotonin depressed tone-evoked responses in most neurons, its effects on responses to FM sweeps were evenly mixed between depression and facilitation. Thus in most cells serotonin had a different effect on tone-evoked responses than it did on FM-evoked responses. In some neurons serotonin depressed responses evoked by tone bursts but left the responses to FM sweeps unchanged, whereas in others serotonin had little or no effect on responses to tone bursts but substantially facilitated responses to FM sweeps. In addition, serotonin could differentially affect responses to various FM sweeps that differed in temporal or spectral structure. Previous studies have revealed that the efficacy of the serotonergic innervation is partially modulated by sensory stimuli and by behavioral states. Thus our results suggest that the population activity evoked by a particular sound is not simply a consequence of the hard wiring that connects the IC to lower and higher regions but rather is highly dynamic because of the functional reconfigurations induced by serotonin and almost certainly other neuromodulators as well. PMID- 10479708 TI - Neuronal interactions improve cortical population coding of movement direction. AB - Interactions among groups of neurons in primary motor cortex (MI) may convey information about motor behavior. We investigated the information carried by interactions in MI of macaque monkeys using a novel multielectrode array to record simultaneously from 12-16 neurons during an arm-reaching task. Pairs of simultaneously recorded cells revealed significant correlations in their trial-to trial firing rate variation when estimated over broad (600 msec) time intervals. This covariation was only weakly related to the preferred directions of the individual MI neurons estimated from the firing rate and did not vary significantly with interelectrode distance. Most significantly, in a portion of cell pairs, correlation strength varied with the direction of the arm movement. We evaluated to what extent correlated activity provided additional information about movement direction beyond that available in single neuron firing rate. A multivariate statistical model successfully classified direction from single trials of neural data. However, classification was consistently better when correlations were incorporated into the model as compared to one in which neurons were treated as independent encoders. Information-theoretic analysis demonstrated that interactions caused by correlated activity carry additional information about movement direction beyond that based on the firing rates of independently acting neurons. These results also show that cortical representations incorporating higher order features of population activity would be richer than codes based solely on firing rate, if such information can exploited by the nervous system. PMID- 10479709 TI - Levels of serotonin in the hemolymph of Aplysia are modulated by light/dark cycles and sensitization training. AB - Serotonin (5-hydroxytryptamine, 5-HT) modulates the behavior and physiology of both vertebrate and invertebrate animals. Effects of injections of 5-HT and the morphology of the serotonergic system of Aplysia indicate that 5-HT may have a humoral, in addition to a neurotransmitter, role. To study possible humoral roles of 5-HT, we measured 5-HT in the hemolymph. The concentration of 5-HT in the hemolymph was approximately 18 nM, a value close to previously reported thresholds for eliciting physiological responses. The concentration of 5-HT in the hemolymph expressed a diurnal rhythm. In addition, electrical stimulation that leads to long-term sensitization significantly increased levels of 5-HT in the hemolymph during training, 1.5 hr after training, and 24 hr after training. Moreover, levels of 5-HT in the hemolymph were significantly correlated with the magnitude of sensitization. The half-life of an increase in 5-HT in the hemolymph was approximately 0.5 hr. Therefore, the persistent increase of 5-HT in the hemolymph 24 hr after sensitization training indicates that training caused a long-lasting increase in the release of 5-HT. This long-lasting increase in 5-HT in the hemolymph was blocked by treatment with an inhibitor of protein synthesis during training. Based on the levels of 5-HT in the hemolymph and its regulation by environmental events, we propose that 5-HT has a humoral role in regulation of the behavioral state of Aplysia. In support of this hypothesis, we found that increasing levels of 5-HT in the hemolymph led to significant alterations in feeding behavior. Increasing levels of 5-HT during the daytime when they were normally low increased the latency to assume feeding posture from daytime to nighttime values. PMID- 10479710 TI - On the synchronizing mechanisms of tetanically induced hippocampal oscillations. AB - gamma (30-100 Hz) and beta (10-30 Hz) oscillations follow tetanic stimulation in the CA1 region of the rat hippocampal slice. Pyramidal neurons undergo a slow depolarization after the tetanus and generate synchronous action potentials. The slow depolarization was previously attributed to metabotropic glutamate receptor (mGluR) activation. However, we found that this event was mediated by GABA(A) receptors, being blocked by bicuculline (50 microM) and accompanied by a dramatic drop in input resistance. Experiments with NMDA and non-NMDA glutamate receptor antagonists revealed that fast synaptic excitation was not necessary for oscillations. IPSPs were strongly depressed during the oscillations. Instead, synchronization was caused by field effects, as shown by: (1) Action potentials of pyramidal neurons proximal (<200 micrometer) to the stimulation site were often preceded by negative deflections of the intracellular potential that masked a net transmembrane depolarization caused by the population spike. (2) Pyramidal neurons located on the surface of the slice, where field effects are weak, fired repetitively but were not synchronized to the network activity. (3) A moderate decrease (50 mOsm) in artificial CSF (ACSF) osmolality did not affect the slow depolarization but increased oscillation amplitude and duration and recruited previously silent neurons into oscillations. (4) 50 mOsm increase in ACSF osmolality dramatically reduced, or abolished, post-tetanic oscillations. Phasic IPSPs, not detectable in proximal neurons, were present, late in the oscillation, in cells located 200-400 micrometer from the stimulation site and possibly contributed to slowing the rhythm during the gamma to beta transition. PMID- 10479712 TI - Sex dimorphisms in the rate of age-related decline in spatial memory: relevance to alterations in the estrous cycle. AB - The present experiments demonstrate the existence of sex differences in the rate of development and the magnitude of age-dependent impairments in cognitive and sensorimotor abilities. Although no sex differences were found in spatial reference memory at a young age, the mnemonic ability of female rats deteriorated more rapidly than that of male rats. A major drop in reference memory of the females occurred at the age of 12 months, whereas in the males the onset of impairments occurred later, at the age of 18 months. In spatial working memory, on the other hand, the magnitude of decline was greater in females than in males, although the onset of these impairments occurred at the age of 24 months in both sexes. A sexual dimorphism-aging interaction also was observed in sensorimotor performance. Up to the age of 18 months the females outperformed the males. Subsequently, by the age of 24 months, the performance of the females declined to a level similar to that of the males. The deficits observed in reference and working memory seem to be cognitive in origin and not attributable to alterations in sensory and motor abilities. In addition, the earlier onset of reference memory impairments in females generally coincides with the onset of alterations in the estrous cycle, suggesting that a decline in the estrogenic milieu of the females could be a factor in accelerating the rate of age-related cognitive impairments in the female rat. PMID- 10479711 TI - Reversals of age-related declines in neuronal signal transduction, cognitive, and motor behavioral deficits with blueberry, spinach, or strawberry dietary supplementation. AB - Ample research indicates that age-related neuronal-behavioral decrements are the result of oxidative stress that may be ameliorated by antioxidants. Our previous study had shown that rats given dietary supplements of fruit and vegetable extracts with high antioxidant activity for 8 months beginning at 6 months of age retarded age-related declines in neuronal and cognitive function. The present study showed that such supplements (strawberry, spinach, or blueberry at 14.8, 9.1, or 18.6 gm of dried aqueous extract per kilogram of diet, respectively) fed for 8 weeks to 19-month-old Fischer 344 rats were also effective in reversing age related deficits in several neuronal and behavioral parameters including: oxotremorine enhancement of K(+)-evoked release of dopamine from striatal slices, carbachol-stimulated GTPase activity, striatal Ca(45) buffering in striatal synaptosomes, motor behavioral performance on the rod walking and accelerod tasks, and Morris water maze performance. These findings suggest that, in addition to their known beneficial effects on cancer and heart disease, phytochemicals present in antioxidant-rich foods may be beneficial in reversing the course of neuronal and behavioral aging. PMID- 10479713 TI - Neurobiological and psychophysical mechanisms underlying the oral sensation produced by carbonated water. AB - Carbonated drinks elicit a sensation that is highly sought after, yet the underlying neural mechanisms are ill-defined. We hypothesize that CO(2) is converted via carbonic anhydrase into carbonic acid, which excites lingual nociceptors that project to the trigeminal nuclei. We investigated this hypothesis using three methodological approaches. Electrophysiological methods were used to record responses of single units located in superficial laminae of the dorsomedial aspect of trigeminal subnucleus caudalis (Vc) evoked by lingual application of carbonated water in anesthetized rats. After pretreatment of the tongue with the carbonic anhydrase inhibitor dorzolamide, neuronal responses to carbonated water were significantly attenuated, followed by recovery. Using c-Fos immunohistochemistry, we investigated the distribution of brainstem neurons activated by intraoral carbonated water. Fos-like immunoreactivity (FLI) was significantly higher in the superficial laminae of dorsomedial and ventrolateral Vc in animals treated with carbonated water versus controls. Dorzolamide pretreatment significantly reduced FLI in dorsomedial Vc. We also examined the sensation elicited by carbonated water in human psychophysical studies. When one side of the tongue was pretreated with dorzolamide, followed by bilateral application of carbonated water, a significant majority of subjects chose the untreated side as having a stronger sensation and assigned significantly higher intensity ratings to that side. Dorzolamide did not reduce irritation elicited by pentanoic acid. The present data support the hypothesis that carbonated water excites lingual nociceptors via a carbonic anhydrase-dependent process, in turn exciting neurons in Vc that are presumably involved in signaling oral irritant sensations. PMID- 10479714 TI - The effects of acute nicotine on the metabolism of dopamine and the expression of Fos protein in striatal and limbic brain areas of rats during chronic nicotine infusion and its withdrawal. AB - The effects of acute nicotine (0.5 mg/kg, s.c.) on dopamine (DA) metabolism and Fos protein expression in striatal and limbic areas of rats on the seventh day of chronic nicotine infusion (4 mg. kg(-1). d(-1)) and after 24 or 72 hr withdrawal were investigated. In saline-infused rats, acute nicotine elevated striatal and limbic 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) concentrations significantly. During the nicotine infusion, no such increases were seen in the striatum, but limbic HVA was somewhat elevated. After 24 hr withdrawal when no nicotine was found in the plasma, acute nicotine elevated striatal DOPAC and HVA and limbic HVA. However, the limbic DOPAC was unaffected. Acute nicotine increased Fos immunostaining (IS) in the caudate-putamen (CPU), the core of nucleus accumbens (NAcc), the cingulate cortex (Cg), and the central nucleus of amygdala (ACe) significantly. During nicotine infusion the nicotine induced responses were attenuated in CPU and NAcc, whereas in ACe and Cg Fos immunostaining was increased as in saline-infused rats. After 24 hr withdrawal, acute nicotine did not increase Fos immunostaining in CPU, NAcc, and Cg, but increased it clearly in ACe. After 72 hr withdrawal, nicotine's effects were restored. Our findings suggest that the nicotinic receptors in the striatal areas are desensitized more easily than those in the limbic areas. Furthermore, the effects of nicotine on various DA metabolites differ. We also found evidence for long-lasting inactivation of nicotinic receptors in vivo regulating limbic dopamine metabolism and Fos expression in striatal and limbic areas. These findings might be important for the protective effects of nicotine in Parkinson's disease and in its dependence-producing properties. PMID- 10479715 TI - Impaired K(+) homeostasis and altered electrophysiological properties of post traumatic hippocampal glia. AB - Traumatic brain injury (TBI) can be associated with memory impairment, cognitive deficits, or seizures, all of which can reflect altered hippocampal function. Whereas previous studies have focused on the involvement of neuronal loss in post traumatic hippocampus, there has been relatively little understanding of changes in ionic homeostasis, failure of which can result in neuronal hyperexcitability and abnormal synchronization. Because glia play a crucial role in the homeostasis of the brain microenvironment, we investigated the effects of TBI on rat hippocampal glia. Using a fluid percussion injury (FPI) model and patch-clamp recordings from hippocampal slices, we have found impaired glial physiology 2 d after FPI. Electrophysiologically, we observed reduction in transient outward and inward K(+) currents. To assess the functional consequences of these glial changes, field potentials and extracellular K(+) activity were recorded in area CA3 during antidromic stimulation. An abnormal extracellular K(+) accumulation was observed in the post-traumatic hippocampal slices, accompanied by the appearance of CA3 afterdischarges. After pharmacological blockade of excitatory synapses and of K(+) inward currents, uninjured slices showed the same altered K(+) accumulation in the absence of abnormal neuronal activity. We suggest that TBI causes loss of K(+) conductance in hippocampal glia that results in the failure of glial K(+) homeostasis, which in turn promotes abnormal neuronal function. These findings provide a new potential mechanistic link between traumatic brain injury and subsequent development of disorders such as memory loss, cognitive decline, seizures, and epilepsy. PMID- 10479716 TI - Activation of cAMP-dependent protein kinase A in prefrontal cortex impairs working memory performance. AB - Activation of the adenylyl cyclase-cAMP-protein kinase A (PKA) intracellular signaling cascade is necessary for long-term memory consolidation in brain regions such as the hippocampus. However, the role of the PKA cascade in the working memory functions of the prefrontal cortex (PFC) is unknown. The present study examined the effects of manipulating PKA activity in the PFC using the cAMP stereoisomers Sp-cAMPS and Rp-cAMPS, which activate and inhibit PKA, respectively. Animals received bilateral infusions of Sp-cAMPS and/or Rp-cAMPS into the PFC immediately before testing on the delayed alternation task, a test of spatial working memory that depends on the integrity of the PFC. Low doses of Sp-cAMPS (0.21, 2. 1, or 21 nmol/0.5 microl) produced a marked, dose-dependent impairment in working memory performance. The impairment produced by infusion of Sp-cAMPS (21 nmol/0.5 microl) was fully reversed by co-infusion of Rp-cAMPS (21 nmol/0.5 microl), consistent with actions on PKA. Rp-cAMPS (21 or 42 nmol/0.5 microl) by itself had no effect on performance. These results indicate that activation of the PKA intracellular signaling cascade in the PFC impairs working memory performance. The current findings contrast with studies of long-term memory consolidation, in which inhibition of PKA with agents such as Rp-cAMPS impaired memory consolidation (Bernabeu et al., 1997; Bourtchouladze et al., 1998), whereas enhancement of the PKA pathway improved memory (Bernabeu et al., 1997; Barad et al., 1998). These results demonstrate that discrete cognitive processes subserved by different cortical regions are mediated by distinct intracellular mechanisms. PMID- 10479717 TI - Axonal transport of activating transcription factor-2 is modulated by nerve growth factor in nociceptive neurons. AB - The aim of this study was to determine whether axonal transport of activating transcription factor-2 (ATF2) occurs in adult sensory neurons, and whether this process is under neurotrophin control. Antisera to both total ATF2 and to the activated (i.e., phosphorylated) form were used for immunocytochemistry and Western blotting. ATF2 was localized to predominantly nociceptive dorsal root ganglion cells in adult rats and shown to accumulate proximal and distal to a sciatic nerve ligature as a result of axonal transport. Subcutaneous injection of nerve growth factor (NGF) decreased the levels of fast retrograde axonal transport of activated ATF2 by 97% (p < 0.05) and elevated levels of retrograde axonal transport of total ATF2 by twofold (p < 0.02). In contrast, blocking endogenous NGF using an anti-NGF antibody induced an elevation in retrograde axonal transport of activated ATF2 of 4. 5-fold (p < 0.05) and decreased retrograde axonal transport of total ATF2 by 72% (p < 0.05). NGF or anti-NGF treatment had no effect on the anterograde transport levels of total or activated ATF2. This study shows that signaling by target-derived NGF to the cell bodies of sensory neurons consists, in part, of the modulation of levels and activation status of a retrogradely transported transcription factor, ATF2. PMID- 10479718 TI - Hot and cold nociception are genetically correlated. AB - Recent experiments in our laboratory have revealed a genetic correlation of the sensitivity of inbred mouse strains to different assays of nociception using noxious heat stimuli. An open question is whether the property of the noxious stimulus underlying the genetic correlation is heat specifically or simply a temperature (hot or cold) in the noxious range. The existing electrophysiological, psychophysical, neurochemical, and functional imaging literatures regarding the relationship of heat pain and cold pain are quite contentious, with a number of similarities and dissociations being documented. In the present study, we tested 12 inbred mouse strains (129/J, A/J, AKR/J, BALB/cJ, C3H/HeJ, C57BL/6J, C57BL/10J, C58/J, CBA/J, DBA/2J, RIIIS/J, and SM/J) for their reflexive withdrawal sensitivity to immersion of the tail in hot (47. 5 degrees C) and cold (-15 degrees C) water and compared the observed latencies with those obtained previously, using a 49 degrees C stimulus. All three traits displayed substantial heritabilities, ranging from 0.41 to 0.50. Strain means on these nociceptive traits displayed a high degree of cross-correlation (r = 0.49-0.77). Genetic correlation of hot and cold nociception implies that similar genes underlie interindividual variability in both traits in mice and further suggests that these nociceptive types share physiological mediation. This finding is discussed in relation to existing data in other mammals including humans. PMID- 10479720 TI - Neocortical synchronized oscillations induced by thalamic disinhibition in vivo. AB - Thalamocortical circuits are recognized as the main elements involved in the genesis of synchronized oscillations typical of certain generalized seizures. We addressed the capability of thalamic disinhibition to generate synchronized oscillations in neocortex. Microdialysis was used to infuse GABA(A) and GABA(B) receptor antagonists directly into the thalamus of anesthetized rats while recording cortical field potentials from 16 sites aligned perpendicular to the cortical surface, using 100 microm spaced linear array silicon probes. The results demonstrate that block of thalamic GABA(A) receptors induces continuous 3 Hz discharges in neocortex and that thalamic GABA(B) receptors mediate this activity. Also, during thalamic disinhibition sporadic long-lasting discharges at 12 Hz occur that do not depend on GABA(B) receptors. Current source density analysis of these activities revealed that the dynamics of sinks and sources for the 3 and 12 Hz discharges was quite distinct, in a way that suggests a different active involvement of the neocortex. The results indicate that intrathalamic inhibitory processes play an essential role in the generation of neocortical synchronized oscillatory activity that may be related to certain forms of generalized seizures. PMID- 10479719 TI - Anatomy of an endogenous antagonist: relationship between Agouti-related protein and proopiomelanocortin in brain. AB - Agouti-related protein (AGRP) is a recently discovered orexigenic neuropeptide that inhibits the binding and action of alpha-melanocyte-stimulating hormone derived from proopiomelanocortin (POMC) at the melanocortin 3 receptor (MC3R) and melanocortin 4 receptor (MC4R) and has been proposed to function primarily as an endogenous melanocortin antagonist. To better understand the interplay between the AGRP and melanocortin signaling systems, we compared their nerve fiber distributions with each other by immunohistochemistry and their perikarya distribution with MC3R and MC4R by double in situ hybridization. Although deriving from distinct cell groups, AGRP and melanocortin terminals project to identical brain areas. Both AGRP and melanocortin neurons selectively express the MC3R, which provides a neuroanatomical basis for a dual-input circuit with biological amplification and feedback inhibition. These studies highlight a broader complexity in POMC-mediated behavior in the brain. PMID- 10479721 TI - Gene expression in individual cells: analysis using global single cell reverse transcription polymerase chain reaction (GSC RT-PCR). AB - The determination of the gene expression pattern of single cells has important implications for many areas of cellular and developmental biology including lineage determination, identification of primitive stem cells and temporal gene expression patterns induced by changes in the cellular microenvironment. Global Single Cell Reverse Transcription-Polymerase Chain Reaction (GSC RT-PCR) enables the study of single cell gene expression patterns. Initial observations of significant heterogeneity among single cells derived from a population of cells prompted us to determine how much of this observed heterogeneity was due to the intrinsic variation within the method. In this paper we discuss the sensitivity of GSC RT-PCR for analysis of differences in gene expression between single cells and, in particular, detail the amount of variation generated by the method itself. We found that most of the intrinsic variation in the method occurred in the PCR step. The total variation induced by the method was in the range of 5 fold. While we have determined that there is a five fold methodological variation in GSC RT-PCR, any method which use its components (including generation of cDNAs for microarray analysis) is likely to be affected by such experimental variability, which could limit the interpretation of the resulting data. PMID- 10479722 TI - Differential sensitivity of double minute chromosomes to hydroxyurea treatment in cultured methotrexate-resistant mouse cells. AB - Treating mammalian cells with continuous sub-lethal doses of Hydroxyurea (HU) causes the loss of double minute chromosomes (DMs) containing amplified oncogenes in culture. Recently, we have shown that treating glioblastoma multiforme cells in culture with low doses of HU causes the loss of DMs containing epidermal growth factor receptor genes. Loss of amplified EGFR genes was accompanied by cessation of growth, and greatly decreased tumorigenicity. To further study HU induced elimination of DMs we have now followed the fate of dihydrofolate reductase gene (DHFR) amplifying DMs in methotrexate-resistant mouse cells during simultaneous treatment with both MTX and HU. We report that in the presence of both HU and MTX, the amplified genes decreased to 25% of starting levels in the first week of treatment, but that ultimately the cells become resistant to HU and reamplify the DHFR gene. We also report that some DHFR amplifying DMs are much more sensitive to HU than others. This study demonstrates that HU does not simply increase the rate of passive loss of DMs. PMID- 10479723 TI - Human transcription factor SLUG: mutation analysis in patients with neural tube defects and identification of a missense mutation (D119E) in the Slug subfamily defining region. AB - Studies in mouse, chicken and Xenopus have shown that Slug is selectively expressed in the dorsal part of the developing neural tube. Ablation and antisense experiments in chicken suggest that Slug may be an important factor during neural tube closure. We therefore investigated the role of Slug as a possible candidate contributing to the aetiology of neural tube defects (NTD) in humans. We characterised the genomic structure of human SLUG including determination of the exon-intron boundaries. The coding sequence of SLUG was screened for mutations in 150 patients with NTD using single strand conformation analysis (SSCA). In one patient, we identified a missense mutation 1548C-->A in exon 2 causing an exchange of a conserved amino acid (D119E) in the Slug subfamily-defining region preceding the first zinc finger. This is the first description of a human mutation in the SLUG gene. In accordance with the findings in model organisms, the SLUG mutation may be causally related to the development of NTD in our patient and could be considered as a predisposing factor. PMID- 10479724 TI - Inactivation of both alleles of the DPC4/SMAD4 gene in advanced colorectal cancers: identification of seven novel somatic mutations in tumors from Japanese patients. AB - Loss of heterozygosity (LOH) of loci on chromosome 18q occurs in a majority of colorectal cancers. The DPC4/SMAD4 gene, lying in close proximity to the DCC gene at 18q21.1, was recently identified as a candidate tumor suppressor for the genesis of pancreatic cancer as well as a predisposing gene for Juvenile Polyposis Syndrome (JPS). The gene product functions as a cytoplasmic mediator in the signaling pathway of transforming growth factor beta (TGF-beta). To investigate the potential role of DPC4/SMAD4 gene in colorectal cancers, we examined 73 tumors of clinical stages II or III from Japanese patients, for LOH at 18q21 and also for subtle mutations anywhere within the coding region of DPC4/SMAD4. LOH was identified in 50 (78%) of the 64 tumors that were informative for polymorphic markers in the region. Somatic mutations were identified in seven of those tumors: two frameshift mutations, a 1-bp deletion (326 del T) in exon 8 and a 1-bp insertion (50-51 ins A) in exon 1; two nonsense mutations, Arg445Ter in exon 10 and Glu538Ter in exon 11; and three missense mutations, Asn129Lys in exon 2, Tyr95Asn in exon 2, and Asp355Glu in exon 8. Three of the seven mutations were observed in the mad homology 1 (MH1) domain encoded by exons 1 and 2. In all of the tumors carrying intragenic mutations of one allele, LOH analysis had shown that the other allele was missing. The results demonstrated that inactivation of both alleles of the DPC4/SMAD4 gene occurs in a substantial proportion of advanced colorectal cancers, and that the DPC4/SMAD4 gene probably exerts a tumor suppressor effect for colorectal carcinogenesis that fulfills the criterion of the two-hit concept proposed by Knudson [A.G. Knudson, Hereditary cancer, oncogenes, and anti-oncogenes, Cancer Res. 45 (1985) 1437-1443.]. PMID- 10479725 TI - Needle-in-a-haystack detection and identification of base substitution mutations in human tissues. AB - Background and induced germline mutagenesis and other genotoxicity studies have been hampered by the lack of a sufficiently sensitive technique for detecting mutations in a small cluster of cells or a single cell in a tissue sample composed of millions of cells. The most frequent type of genetic alteration is intragenic. The vast majority of oncogenic mutations in human and mammalian cancer involves only single base substitutions. We have developed universally applicable techniques that not only provide the necessary sensitivity and specificity for site specific mutagenesis studies, but also identify the point mutation. The exponential amplification procedures of polymerase chain reaction (PCR) and ligase chain reaction (LCR) have been combined with restriction endonuclease (RE) digestion to enable the selective enrichment and detection of single base substitution mutations in human oncogenic loci at a sensitivity of one mutant in more than 10(7) wild type alleles. These PCR/RE/LCR procedures have been successfully designed and used for codons 12 and 248 of the Ha-ras and p53 genes, respectively, both of which contain a natural MspI restriction endonuclease recognition sequence. These procedures have also been adapted for the detection and identification of mutations in oncogenic loci that do not contain a natural restriction endonuclease recognition sequence. Using PCR techniques, a HphI site was incorporated into the codons 12/13 region of the human N-ras gene, which was then used for the selective enrichment of mutants at this oncogenic locus. These PCR/RE/LCR procedures for base substitution mutations in codon 12 of the N-ras gene were found to have the sensitivity of detection of at least one mutant allele in the presence of the DNA equivalent of 10(6) wild type cells. Only one peripheral blood leukocyte DNA specimen out of nine normal individuals displayed an observable Ha-ras mutation that was present at frequency between 10(-5) and 10(-6). These PCR/RE/LCR techniques for detecting and identifying base substitution mutations are universally applicable to almost any locus or base site within the human or animal genome. With the added advantage of the adjustability of both the amount of DNA (number of genomes) to be tested and the sensitivity (10(-2) to 10(-7)) of the assay selection or enrichment procedures, these PCR/RE/LCR techniques will be useful in addressing a broad range of important questions in mutagenesis and carcinogenesis. PMID- 10479726 TI - A characterization of genetic variants in BRCA1 intron 8 identifies a mutation and a polymorphism. AB - The biochemical and genetic characterizations of two variants that occur in BRCA1 intron 8 are presented. The variant IVS8+2T-->C induces an aberrant transcript that deletes exon 8. This exon-skipping deletion disrupts the open reading frame by juxtaposing exon 7 and exon 9 in the aberrant splice product. Theoretically, 50 abnormal residues from reading frame 2 are translated following exon 7 before a stop codon is encountered. The chromosomal contribution to the relevant RNA species was tracked using a silent polymorphism at codon 694 (serine AGC or AGT). Nucleotide sequencing of this polymorphic codon demonstrated that the aberrant transcript was derived solely from the chromosome encoding AGT. The normally spliced productive transcript also displayed loss of heterozygosity and was derived solely from the chromosome encoding AGC at codon 694. Also, a haplotype analysis using a breast cancer patient database showed that the chromosome bearing serine 694-AGT carried IVS8+2T-->C. A second more common variant, IVS8 58delT, was characterized as a polymorphism. Analysis of RNA from patient samples used the same silent polymorphism at codon 694 and showed that the normal message was derived from both chromosomes. PMID- 10479727 TI - Restriction fragment analysis as a source of error in detection of heteroplasmic mtDNA mutations. AB - The transition from A to G at nt 5656 (5656A-->G) in mitochondrial DNA has been suggested to be a pathogenic mutation and, furthermore, a heteroplasmic one. We found that the mutation was present in 14 out of 83 healthy controls from northern Finland and that 5656A-->G was exclusively associated with mtDNA haplogroup U. Interestingly, 5656A-->G appeared to be heteroplasmic in NheI digestion of PCR fragments that were amplified by using a mismatched oligonucleotide primer creating a digestion site in the presence of the mutant variant. However, we did not detect the wild type genome in clones from such a sample and subsequent experiments revealed that the apparent heteroplasmy was due to inhibition of NheI by NaCl. Our results suggest that 5656A-->G is a polymorphism and it may be highly characteristic for Finns. Furthermore, new heteroplasmic mutations identified by restriction fragment analysis should be adequately controlled for any false positive results that may be due to incomplete digestion. PMID- 10479728 TI - Polymorphisms in the human DNA repair gene XPF. AB - DNA sequence polymorphisms were sought in the coding region and at the exon intron boundaries of the human XPF gene, which plays a role in nucleotide excision repair. Based on a survey of 38 individuals, we found six single nucleotide polymorphisms, one in the 5' non-coding region of the XPF gene, and five in the 2751 bp coding region. At each site, the frequency of the rarer allele varies from about 0.01 to over 0.38. Except for the 5' non-coding and one coding sequence polymorphism, the rarer alleles for the remaining four polymorphisms were found only in heterozygotes. Of the five polymorphisms in the coding region, one is silent, one results in a conserved amino acid difference, and the remaining three result in non-conserved amino acid differences. Because of its biological function in nucleotide excision repair, functionally significant XPF gene polymorphisms are candidates for influencing cancer susceptibility and overall genetic stability. Nucleotide sequence diversity estimates for XPF are similar to the lipoprotein lipase and beta-globin genes. PMID- 10479729 TI - Screening of the human tumor necrosis factor-alpha (TNF-alpha) gene promoter polymorphisms by PCR-DGGE analysis. AB - We have designed a new PCR-DGGE technique that enables detection of base changes in the TNF-alpha gene promoter. Screening of 130 samples from Spanish children has shown that this technique accurately detects the altered band patterns induced by the presence of the polymorphisms at positions -376, -308, -238 and 163 of the promoter sequence. Although further analysis are needed to fully characterise the alterations detected, we believe that this PCR-DGGE technique is a rapid and sensitive first approach to the genetic characterisation of the TNF alpha promoter. PMID- 10479730 TI - The hydration of nucleic acid duplexes as assessed by a combination of volumetric and structural techniques. AB - Using high precision densimetric and ultrasonic measurements, we have determined, at 25 degrees C, the apparent molar volumes PhiV and the apparent molar compressibilities PhiK(S) of four nucleic acid duplexes-namely, the DNA duplex, poly(dIdC)poly(dIdC); the RNA duplex, poly(rA)poly(rU); and the two DNA/RNA hybrid duplexes, poly(rA)poly(dT) and poly(dA)poly(rU). Using available fiber diffraction data on these duplexes, we have calculated the molecular volumes as well as the solvent-accessible surface areas of the constituent charged, polar, and nonpolar atomic groups. We found that the hydration properties of these nucleic acid duplexes do not correlate with the extent and the chemical nature of the solvent-exposed surfaces, thereby suggesting a more specific set of duplex water interactions beyond general solvation effects. A comparative analysis of our volumetric data on the four duplexes, in conjunction with available structural information, suggests the following features of duplex hydration: (a) The four duplexes exhibit different degrees of hydration, in the order poly(dIdC)poly(dIdC) > poly(dGdC)poly(dGdC) > poly(dAdT)poly(dAdT) approximately poly(dA)poly(dT). (b) Repetitive AT and IC sequences within a duplex are solvated beyond general effects by a spine of hydration in the minor groove, with this sequence-specific water network involving about 8 additional water molecules from the second and, perhaps, even the third hydration layers. (c) Repetitive GC and IC sequences within a duplex are solvated beyond general effects by a "patch of hydration" in the major groove, with this water network involving about 13 additional water molecules from the second and, perhaps, even the third hydration layers. (d) Random sequence, polymeric DNA duplexes, which statistically lack extended regions of repetitive AT, GC, or IC sequences, do not experience such specific enhancements of hydration. Consequently, consistent with our previous observations (T. V. Chalikian, A. P. Sarvazyan, G. E. Plum, and K. J. Breslauer, Biochemistry, 1994, Vol. 33, pp. 2394-2401), duplexes with approximately 50% AT content exhibit the weakest hydration, while an increase or decrease from this AT content causes enhancement of hydration, either due to stronger hydration of the minor groove (an increase in AT content) or due to stronger hydration of the major groove (an increase in GC content). (e) In dilute aqueous solutions, a B DNA duplex is more hydrated than an A-DNA duplex, a volumetric-based conclusion that is in agreement with previous results obtained on crystals, fibers, and DNA solutions in organic solvent-water mixtures. (f) the A-like, RNA duplex poly(rA)poly(rU) and the structurally similar A-like, hybrid duplex poly(rA)poly(dT), exhibit similar hydration properties, while the structurally distinct A-like, hybrid duplex poly(rA)poly(dT) and non-A-like, hybrid duplex poly(dA)poly(rU) exhibit differential hydration properties, consistent with structural features dictating hydration characteristics. We discuss how volumetric characterizations, in conjunction with structural studies, can be used to describe, define, and resolve the general and sequence/conformation-specific hydration properties of nucleic acid duplexes. PMID- 10479731 TI - Interaction between fatty acid salts and elastin: kinetics, absorption equilibrium, and consequences for elasticity. AB - Elastin from bovine ligamentum nuchae is incubated in aqueous solutions of sodium salts of fatty acids (FAS). The FAS are laurate, myristate, and palmitate. Absorption of FAS in the elastin network is studied as a function of time, FAS concentration, and ionic strength. The consequences of this uptake for the elasticity of the elastin are studied by static and dynamic stress-strain measurements. Generally, distinction must be made between the initial time dependent stage (I) and the final equilibrium stage (II). In I the initial rate of absorption follows a second-order binding mechanism, with the rate constant increasing with decreasing length of the FAS. In this regime, the elasticity modulus remains more or less unaffected. Especially in regime II the absorption of FAS is enhanced by a reduction in the cross-link density in the elastin network. This is ascribed to an osmotic pressure primarily caused by the concomitant uptake of low molecular weight ions in the elastin. The absorption equilibrium can be described by Langmuir theory. The absorption affinity increases with increasing hydrocarbon chain length of the FAS, indicating the contribution of hydrophobic interaction. Although the elasticity is not lost, the modulus is now reduced and a concomitant viscous component is developed. PMID- 10479732 TI - Examination of the structure/function relationship in the exchangeable apolipoprotein, apolipophorin-III. AB - Exchangeable apolipoproteins are proteins that reversibly bind lipoprotein particles to facilitate their transport in vivo. The structure/function relationship of apolipophorin-III (apo-III), the only insect exchangeable apolipoprotein, has been investigated by examining the association of this protein with lipid vesicles. The importance of a conserved leucine residue, reported to be essential for apo-III binding to lipids, has been evaluated through site-directed mutagenesis. A unique cysteine replaces the conserved leucine at position 30 in recombinant apo-III (L30C protein). This substitution results in the covalent dimerization of the apo-III mutant via a disulfide bond. The cysteine mutation causes no difference in surface hydrophobicity of the L30C proteins when compared to the wild type apo-III. Wild type apo-III, L30C monomer, and L30C dimer associate with dimyristoylphosphatidylcholine (DMAC) vesicles in a similar manner, resulting in a reduction of turbidity of a phospholipid vesicle suspension. Analysis with transmission electron microscopy (TEM) reveals disk like complexes identical to those previously reported with the wild type apo-III. Because the mutation of the conserved leucine seems to affect the solution behavior and surface hydrophobicity of apo-III, this residue is likely to be exposed to the aqueous environment. However, the similar behaviors of the wild type protein, the L30C monomer, and L30C dimer with respect to the binding of phospholipid vesicles suggest that this residue is not absolutely required for the protein binding to hydrophobic or amphiphilic interfaces. PMID- 10479733 TI - Exploring the (1 --> 3)-beta-D-glucan conformational phase diagrams to optimize the linear to macrocycle conversion of the triple-helical polysaccharide scleroglucan AB - The immunologically important (1 --> 6) comb-like branched (1 --> 3)-beta-D glucans scleroglucan, schizophyllan, lentinan, and others, exist mainly as linear triple-helical structures in aqueous solution. Partial interconversion from linear to circular topology has been reported to take place following conformational transition of the triple-helical structure and subsequent regeneration of the triplex conformation. We here report on experimental data indicating that complete strand separation of the triple-helical structure is required for this interconversion. NaOH or dimethylsulfoxide was used to induce dissociation of the triplex at combinations of concentrations and temperatures shown by calorimetry to yield a conformational transition of the triplex structures. For the alkaline treatment at 55 degrees C, it is found that up to about 30% of the material readily can be converted to the cyclic topology. This fraction increased to about 60% when the subsequent annealing of the scleroglucan in aqueous solution at pH 7 was carried out at 100 degrees C. Further increase of the annealing temperature yielded a smaller relative amount of cyclic species. The data indicate that the lower molecular weight fraction of the molecular weight distributions can be converted selectively to the macrocyclic topology by conditions that do not yield complete strand separation of the whole sample. These findings add to previous reports by providing more details about how the conditions required for the linear triplex to macrocycle interconversion relate to the conformational properties of the triple-helical structure. Copyright 1999 John Wiley & Sons, Inc. PMID- 10479734 TI - 600 ps molecular dynamics reveals stable substructures and flexible hinge points in cAMP dependent protein kinase. AB - Molecular dynamics simulations of the catalytic subunit of cAMP dependent protein kinase (cAPK) have been performed in an aqueous environment. The relations among the protein hydrogen-bonding network, secondary structural elements, and the internal motions of rigid domains were examined. The values of fluctuations of protein dihedral angles during dynamics show quite distinct maxima in the regions of loops and minima in the regions of alpha-helices and beta-strands. Analyses of conformation snapshots throughout the run show stable subdomains and indicate that these rigid domains are constrained during the dynamics by a stable network of hydrogen bonds. The most stable subdomain during the dynamics was in the small lobe including part of the carboxy-terminal tail. The most significant flexible region was the highly conserved glycine-rich loop between beta strands 1 and 2 in the small lobe. Many of the main chain dihedral angle changes measured in a comparison of the crystallographic structures of "open" and "closed" conformations of cAPK correspond to the highly flexible residues found during dynamics. PMID- 10479735 TI - Conformational studies of parathyroid hormone (PTH)/PTH-related protein (PTHrP) point-mutated hybrids. AB - The N-terminal 1-34 segments of both parathyroid hormone (PTH) and parathyroid hormone-related protein (PTHrP) bind and activate the same membrane receptor in spite of major differences between the two hormones in their amino acid sequence. Recently, it was shown that in (1-34)PTH/PTHrP segmental hybrid peptides, the N terminal 1-14 segment of PTHrP is incompatible with the C-terminal 15-34 region of PTH leading to substantial reduction in potency. The sites of incompatibility were identified as positions 5 in PTH and 19 in PTHrP. In the present paper we describe the synthesis, biological evaluation, and conformational characterization of two point-mutated PTH/PTHrP 1-34 hybrids in which the arginine residues at positions 19 and 21 of the native sequence of PTHrP have been replaced by valine (hybrid V(21)) and glutamic acid (hybrid E(19)), respectively, taken from the PTH sequence. Hybrid V(21) exhibits both high receptor affinity and biological potency, while hybrid E(19) binds weakly and is poorly active. The conformational properties of the two hybrids were studied in aqueous solution containing dodecylphosphocholine (DPC) micelles and in water/2,2, 2-trifluoroethanol (TFE) mixtures. Upon addition of TFE or DPC micelles to the aqueous solution, both hybrids undergo a coil-helix transition. The maximum helix content in 1 : 1 water/TFE, obtained by CD data for both hybrids, is approximately 80%. In the presence of DPC micelles, the maximum helix content is approximately 40%. The conformational properties of the two hybrids in the micellar system were further investigated by combined 2D-nmr, distance geometry (DG), and molecular dynamics (MD) calculations. The common structural motif, consisting of two helical segments located at N- and C-termini, was observed in both hybrids. However, the biologically potent hybrid V(21) exhibits two flexible sites, centered at residues 12 and 19 and connecting helical segments, while the flexibility sites in the weakly active hybrid E(19) are located at position 11 and in the sequence 20-26. Our findings support the hypothesis that the presence and location of flexibility points between helical segments are essential for enabling the active analogs to fold into the bioactive conformation upon interaction with the receptor. PMID- 10479736 TI - A method to detect nonproline cis peptide bonds in proteins. AB - Based on the geometrical parameters around seventeen incorrectly assigned trans conformations of peptide bonds in protein structures and their correct cis counterparts, we have devised an algorithm that is capable of detecting these sites. The algorithm was optimized to reliably find all of the seventeen test cases. It can be used to quickly scan an atomic coordinate file or the complete Brookhaven Protein Data Base for more likely candidates for non-Pro cis peptide bonds. Also, it can be of help to guide the crystallographer in intermediate stages of structure determination towards suspect areas. PMID- 10479737 TI - Design, synthesis, and conformational studies of the hGM-CSF derived peptide (13 27)-Gly-(75-87). AB - An analogue of the human granulocyte-macrophage colony-stimulating factor (hGM CSF), hGM-CSF(13-27)-Gly-(75-87) was synthesized by solid phase methodology. This analogue was designed to comprise helices A and C of the native growth factor, linked by a glycine bridge. Helices A and C form half of a four-helix bundle motif in the crystal structure of the native factor and are involved in the interaction with alpha- and beta-chains of the heterodimeric receptor. A conformational analysis of the synthetic analogue by CD, two-dimensional nmr spectroscopy, and molecular dynamics calculations is reported. The analogue is in a random structure in water and assumes a partially alpha-helical conformation in a 1 : 1 trifluoroethanol/water mixture. The helix content in this medium is approximately 70%. By 2D-nmr spectroscopy, two helical segments were identified in the sequences corresponding to helices A and C. In addition to medium- and short-range NOESY connectivities, a long-range cross peak was found between the Cbeta proton of Val(16) and NH proton of His(87) (using the numbering of the native protein). Experimentally derived interproton distances were used as restraints in molecular dynamics calculations, utilizing the x-ray coordinates as the initial structure. The final structure is characterized by two helical segments in close spatial proximity, connected by a loop region. This structure is similar to that of the corresponding domain in the x-ray structure of the native growth factor in which helices A and C are oriented in an antiparallel fashion. The N-terminal residues Gly-Pro of helix C are involved in an irregular turn connecting the two helical segments. As a consequence, helix C is appreciably shifted and slightly rotated with respect to helix A compared to the x-ray structure of the native growth factor. These small differences in the topology of the two helices could explain the lower biological activity of this analogue with respect to that of the native growth factor. PMID- 10479738 TI - The study of the conformation and interaction of two tachykinin peptides in membrane mimicking systems by NMR spectroscopy and pulsed field gradient diffusion. AB - Pulsed-field gradient diffusion has been used to study the binding of two tachykinin peptides, [Tyr(8)]-substance P (SP) and [Tyr(0)]-neurokinin A (NKA) to two membrane-mimicking micelles, dodecylphosphocholine, and sodium dodecylsulfate. The structure of these peptides bound to the micelles have also been studied by using two-dimensional nmr and restrained simulated annealing calculations. No major difference in the structures of each peptide in the two micellar media was found. The difference between the micelle-bound structure of [Tyr(8)]SP and that of SP was also minor. The longer helical conformation on the C-terminus for [Tyr(0)]NKA was observed, compared with that for NKA. The relationship between the difference in the biological potencies of [Tyr(8)]SP and SP and the differences in their structure, especially the interaction of the side chains of the two aromatic residues, and the difference in their binding affinities to membrane was discussed. In addition, differences between the result of restrained molecular dynamics simulations of [Tyr(8)]SP in the presence of an explicit micelle and the present results were observed and discussed. PMID- 10479739 TI - The current status of apo E4 as a risk factor for Alzheimer's disease: an epidemiological perspective. PMID- 10479740 TI - The stress and psychological morbidity of the Alzheimer patient caregiver. AB - BACKGROUND: Alzheimer's family caregivers suffer from psychological and physical morbidity associated with the care of the Alzheimer's patient. The objectives of the study were to compare stress and psychological morbidity between family caregivers of Alzheimer's disease patients and family caregivers of aged non demented chronically ill patients and to explore the impact of cognitive and non cognitive patient symptoms on caregivers. METHOD: Fifty-eight caregivers of Alzheimer's patients (cases) and 32 caregivers of non-demented chronically ill patients (controls) were studied. A cross-sectional case-control design was employed using validated scales to assess patient symptomatology and self-report questionnaires to measure stress and psychological morbidity in caregivers. RESULTS: Compared to controls, stress and psychological morbidity were higher in caregivers of Alzheimer's patients. Behavioral symptoms and impairment in instrumental activities of daily living were associated with caregiver stress. Length of care was associated with caregiver psychological morbidity. CONCLUSIONS: Caregiver stress may be reduced with special attention paid to the treatment and management of behavioral and functional symptoms in the Alzheimer's disease patient. Caregiver education and coping skills should also be included. PMID- 10479741 TI - Depression in patients with Parkinson's disease. AB - Parkinson's disease (PD) is a disabling neurodegenerative condition commonly complicated by the existence of comorbid depression. The prevalence rates of depression in this patient group have been reported to be as high as 40%. Currently, depression in PD is undertreated; there have been few controlled clinical trials of antidepressants in this patient group. Patients with PD are usually elderly and often administered a range of medication, therefore the choice of antidepressant must be undertaken with care. Tricyclic antidepressants (TCAs) have been studied in patients with PD and comorbid depression; however, the risk of anticholinergic side-effects means that their use is largely avoided. Selective serotonin reuptake inhibitors have comparable efficacy to the TCAs and a better tolerability profile in patients with depression; they are rapidly being considered as first-line therapy for PD patients with depression. Clinical studies in this patient group are warranted. This article reviews the characteristics of comorbid depression in patients with PD and discusses the treatment options available. PMID- 10479742 TI - 'Yes' ifs, ands or buts: examining performance and correlates of the repetition task in the mini-mental state examination. AB - The aim of this study was to examine whether the type of sentence used in the repetition task included in the Mini-Mental State Examination (MMSE) affected performance in a group of 79 demented and 19 non-demented Hebrew-speaking elderly persons. The cognitive functioning of the participants was assessed using the MMSE and CAMCOG examinations. The performance of the repetition task was evaluated by using three sentences: the literal translation of the English language expression used in the original MMSE; a well-known Hebrew proverb consisting of monosyllabic words and rhythmic effects; and another well-known Hebrew proverb without such attributes. Only a third of the participants successfully repeated the literally translated expression. It showed low predictive value and was highly affected by education. The well-known Hebrew monosyllabic proverb showed moderate predictive value but no discriminatory ability. The other well-known Hebrew proverb performed the best. The translation of the repetition task in the MMSE to other languages is problematic. Strict adherence to the original language proved to be the least desirable choice. PMID- 10479743 TI - Gender, isolation, work patterns and stress among old age psychiatrists. AB - OBJECTIVE: To investigate work patterns and stress in consultant old age psychiatrists over a period of 1 week and to attempt to identify areas amenable to change. DESIGN: Postal survey. PARTICIPANTS: Full-time old age psychiatrists on the list held by the Faculty of Old Age Psychiatry, Royal College of Psychiatrists. MAIN OUTCOME MEASURES: Hours spent on different work activities during a period of 1 week, Stress Checklist score, stressors perceived by respondents. MAIN RESULTS: Men and women doctors did not differ in numbers of hours worked. Consultants working with colleagues worked similar hours to single handed consultants. Community activity was greater among consultants working with colleagues than among single-handed doctors, whose work was based more in outpatient clinics. For the whole group, time in acute ward rounds correlated positively with stress score and time spent on research at home correlated negatively with stress score. CONCLUSIONS: Doctors spending more than 50 hours at work might be advised to review their work pattern. Excessive time on administrative activities should be reduced. Peer support should be encouraged. Consultants and managers should be sensitive to work patterns and possible sources of stress. PMID- 10479744 TI - Can a brief intervention have a longer-term benefit? The case of the research nurse and depressed older people in the community. PMID- 10479745 TI - Prevalence and correlates of depression among Saudi elderly. AB - OBJECTIVES: To assess the prevalence of depression and associated factors in elderly people in Saudi Arabia. METHOD: A cross-sectional national survey of the elderly population of Saudi Arabia was conducted between January 1994 and December 1995. A stratified two-stage sampling technique was used to select the study subjects. In the first stage, a random sample of primary health care centres (PHCs) was selected in each of the five administrative regions of the country. The family health records (FHR) of each selected PHC were reviewed and a list of the elderly (60 years and over) was compiled. In the second stage, a sample of the subjects was selected from the FHR and contacted for a personal interview. The subjects' physical, social and environmental health status were assessed by an interview during which a structured questionnaire was completed. The Geriatric Depression Scale (GDS) was used to screen for depression. Other items of information in the questionnaire included subjects' sociodemographic characteristics, activities of daily living (ADL), laboratory and radiological investigations. RESULTS: The total number of elderly subjects included in this study was 7970. Their mean age+/-standard deviations was 68.8+/-7.7 (male 69.1+/ .7.7; and female 67.7+/-7.5) years. Depressive symptoms were reported in 3110 (39%) of the subjects, and 8.4% were in the severe depressive symptoms score group. Personal characteristics that correlated strongly with depression were poor education (p=0.001), unemployment (p=0.001), divorced or widowed status (p=0.001), old age and being a female (p=0.001). Living in a remote rural area with poor housing arrangements and limited accessibility within the house and poor interior conditions were also significantly associated with high depressive symptoms (p=0.001). Limited privacy, such as having a particular room specified for the elderly, was associated with more depressive symptoms than sharing a room with another person (p=0. 001). Lower incomes inadequate for personal needs as well as depending on charity or other relatives were associated with more cases of depression (p=0.001). The proportion of cases of depression correlated positively with the number of medical diagnoses and medications received (p=0.001). Significant depression was associated with loss of a close relative, living alone and limited participation in recreational activities. Perception of poor health and dependence on others for daily activities were associated with more depressive symptoms (p=0.001). Also health problems, especially faecal or urinary incontinence, were associated with more depressive symptoms (p=0.001). CONCLUSION: Depressive symptoms are common among Saudi elderly. Detection and management rates were low. Primary care teams could help these patients if properly trained. A simple instrument such as the Geriatric Depression Scale is useful and easily administered. PMID- 10479746 TI - Survival in a population-based cohort of dementia patients: predictors and causes of mortality. AB - OBJECTIVES: To examine predictors of survival time and causes of mortality in a population-based cohort of demented subjects. DESIGN: Longitudinal naturalistic follow-up study. SETTING: A rural area in The Netherlands. PARTICIPANTS: The study population consisted of 102 demented subjects derived from a population based, two-stage prevalence study of dementia among subjects aged 65 and over. MEASUREMENTS AND ANALYSIS: Survival analysis was performed using the Realized Probability of Dying as a measure of survival time. Adjustment for duration and severity at study entry was employed in all analyses. RESULTS: A substantial excess mortality was present among demented subjects as compared with their non demented birth cohort. Disorders related to dementia, such as cachexia, dehydration and pneumonia, were major causes of death. Variables related to severity of functional impairment, aphasia and a shorter duration at study entry predicted a shorter survival in the study population. PMID- 10479747 TI - A programme for short-term withdrawal from benzodiazepines in geriatric hospital inpatients: success rate and effect on subjective sleep quality. AB - OBJECTIVE: We tested the hypothesis that a short-term programme for withdrawal of benzodiazepines (BZD) is feasible in hospitalized geriatric patients. METHODS: Fifty-six geriatric subjects who had been taking BZD for at least 3 months were asked to discontinue these drugs upon admission to the inpatient ward. A withdrawal programme including initial substitution therapy combined with psychological consulting was offered. The usual BZD medication was replaced by either lormetazepam 1 mg or trazodone 50 mg, administered at bedtime. After 1 week of replacement therapy all sedative medication was stopped. The subjective estimations of sleep quality were evaluated four times during a period of 6 weeks. RESULTS: Forty-nine patients agreed to participate. In this group four subjects (8.2%) resumed BZD use while in the hospital and another seven subjects (14.3%) relapsed after discharge. Therefore, the overall success rate was 77.6% in the group of volunteers and 67. 9% in the total group of eligible patients. The data of the present study further demonstrate that no major withdrawal symptoms occurred and that the subjective quality of sleep remained virtually unchanged in the course of the programme. The sleep quality was not significantly different in patients on trazodone versus patients on lormetazepam. The success rate was similar in both drug substitution groups. CONCLUSIONS: Short-term withdrawal of BZD may be achieved in two-thirds of elderly hospital inpatients without deterioration of sleep quality or other deleterious side-effects. PMID- 10479748 TI - Factors associated with vascular dementia in an elderly community population. AB - OBJECTIVE: Examination of associated factors for vascular dementia in comparison with Alzheimer's disease and a non-demented control population. SETTING: A semi rural geographically defined population, in Melton Mowbray and the surrounding area, consisting of elderly patients aged 75 and over from a 13-doctor general practice. METHOD: 438 subjects, sampled according to results of initial screening with the Mini-Mental State Examination (MMSE), were examined using the Cambridge Mental Disorders of the Elderly Examination (CAMDEX), which was also used to classify the subtypes of dementia. RESULTS: 34 subjects had vascular dementia, 222 had Alzheimer's disease and 140 were non-demented according to the clinician's diagnosis based on the CAMDEX criteria. Significant factors associated with vascular dementia, when compared to the Alzheimer's disease and non-demented groups, were family history of stroke and diabetes. Stroke was also significant, but this factor is one of those defining the criteria for diagnostic grouping. There were no significant differences between groups for falls, heart attack, hypertension, head injury, substance abuse or lipid levels. CONCLUSION: The results are broadly consistent with the published literature and emphasize the importance of diabetes as a key risk factor for vascular dementia. PMID- 10479749 TI - The impact of two changes in service delivery on a geriatric psychiatry liaison service. AB - BACKGROUND: The impact of two changes in service delivery (alteration in the admission policy of the medical unit and the introduction of a formal liaison component to a 'consultation only' liaison geriatric psychiatry service) on the associated liaison geriatric psychiatry service was examined in a naturalistic study. The main null hypothesis was that the referral rate would not be influenced by either change in service delivery. METHOD: The 30-month study period was divided into three phases defined by the two changes in service delivery. The impact of the two changes in service delivery on the (i) referral rate, (ii) reasons for referral, (iii) demographic and clinical characteristics of the referrals and (iv) advice offered after assessment was examined with a retrospective design. RESULTS: There was a significant decline in the number of referrals for each month across the three study phases exclusively accounted for by a decline between phase 2 and phase 3. The number of reasons for referral and management advice offered for social treatments per patient were greater in phase 1 compared to phase 2. The total number of management advice offered for medical treatments per patient was greater in phase 2 compared to phase 3. The cost of a specialist registrar attending a geriatric medicine ward round was more than offset by the associated decline in the number of referrals per month. CONCLUSION: Although this study was not designed as a cost-effectiveness study, the results suggest that the liaison component has the potential to be cost effective. There is a need for more formal clinical effectiveness and cost effectiveness studies in liaison geriatric psychiatry. PMID- 10479750 TI - 'Not knowing where I am doesn't mean I don't know what I like': cognitive impairment and quality of life responses in elderly people. AB - OBJECTIVE: To elucidate the extent to which elderly people with cognitive impairment are able to answer questions about their quality of life.Design and setting308 elderly residents were interviewed within 2 weeks of admission to one of 30 residential or nursing homes in north-west England. MEASURES: The Mini Mental State Examination (MMSE), the Lancashire Quality of Life Profile (Residential) (LQOLP(R)), the Crichton Royal Behaviour Rating Scale (CRBRS) and the HONOS-65+. RESULTS: Of the 308 subjects, LQOLP(R) interviews were attempted with 213 who scored 10 or over on the MMSE. Of the 213, 77.5% were found to be 'interviewable', ie able to answer the majority of questions in the LQOLP(R) and in doing so to give answers in which the interviewer had confidence. Of the seven cognitive domains measured by the MMSE, visual construction and registration were not significantly associated with interviewability. While orientation to time and recall were significantly associated with interviewability, many interviewable respondents had poor scores in these domains. No respondents were interviewable who scored less than 2 (out of 5) for orientation to place or less than 3 (out of 8) for language or less than 2 (out of 5) for attention. CONCLUSIONS: A high proportion of elderly people can answer questions about their quality of life, even in the presence of significant cognitive deficits. PMID- 10479751 TI - Burden in carers of dementia patients: higher levels in carers of younger sufferers. AB - A comparative study of the clinical characteristics of early onset dementia (onset before the age of 65) and late onset dementia sufferers was conducted. The level of carer burden among carers of both groups was measured. Carer burden was significantly higher among carers of younger sufferers, although there were no significant differences between the groups in terms of clinical features such as severity of dementia or behavioural disturbance. Higher carer burden showed an inverse relationship to patient's age. Our results have implications in terms of encouraging community care for dementia sufferers, and suggest that further exploration to identify factors associated with burden in younger carers is required. PMID- 10479752 TI - UK Alzheimer's disease genetics consortium. PMID- 10479753 TI - Pharmacological basis for the use of the antivenene water soluble extract of Diodia scandens as a laxative, oxytocic agent and a possible aphrodisiac in traditional medicine practice in eastern Nigeria. AB - The effects of the antivenene fraction of an ethanol extract of Diodia scandens on some mammalian smooth muscles were investigated. On the guinea-pig ileum, the extract was shown to be a partial agonist acting via muscarinic receptors. Acetylcholine (ACh) was 2.5 x 10(5) times more potent. On the pregnant guinea-pig uterus, the extract induced concentration-dependent increases in the force of contraction and tonus. Oxytocin and ergometrine were respectively 10(6) and 10(3) times more potent. The extract, at subliminal concentrations, potentiated ACh and adrenaline-induced contractions in the guinea-pig was deferens. It also induced dose-related vasodilatation in the rat hindquarters and depressed the blood pressure in the anaesthetized cat. It was concluded that these pharmacological actions offer some scientific explanation for the use of Diodia scandens in traditional medicine as a laxative and as an oxytocic agent. It is suggested that the extract could enhance erection and ejaculatory processes in the male, thus accounting for its regular use by some elderly males. PMID- 10479754 TI - Triptolide is more effective in preventing T cell proliferation and interferon gamma production than is FK506. AB - The Chinese traditional herb Tripterygium wilfordii Hook. F (TWHF) has been reported to be effective in the treatment of a variety of autoimmune disorders. The major active component of this herb is triptolide and most of the efficacy of this herb immunosuppression is attributed to triptolide. FK506 is also a potent immunosuppressive agent and is currently being used clinically. The present studies compare the effectiveness of triptolide and FK506 to suppress certain human T cell functions. Specifically human T cell proliferation, IL-2 and IFNgamma were compared. The results show that, overall, triptolide is more effective at inhibiting T cell proliferation and IFNgamma production than FK506 and the two compounds inhibit IL-2 production in an equivalent manner. PMID- 10479755 TI - Relationship between the antidiarrhoeal effects of Hange-Shashin-To and its active components. AB - This study was designed to examine the relationship between the antidiarrhoeal effects of Hange-Shashin-To (TJ-14) and its active components. Oral treatment with TJ-14 at 1000 mg/kg significantly inhibited castor oil-induced diarrhoea. Both the 50% methanol eluate fraction (fraction III) and the methanol eluate fraction (fraction IV) showed antidiarrhoeal effects at oral doses of 68 mg/kg and 63 mg/kg, respectively, corresponding to 1000 mg/kg of TJ-14. TJ-14 (1000 mg/kg, p.o.) showed a significant increase in blood corticosterone levels. Increased blood corticosterone was noted after the oral administration of 63 mg/kg of fraction IV. The inhibitory activity of TJ-14 on cyclooxygenase-2 (COX 2) was also observed in fractions III and IV. The main component of fraction III was Scutellariae Radix-derived baicalin. Fraction IV contained Glycyrrhizae Radix derived glycyrrhizin and isoliquiritin, Coptidis Rhizoma-derived berberine, coptisine and palmitine. Ginseng Radix-derived saponins were also present in fraction IV. These compounds inhibited castor-oil induced diarrhoea at oral doses of 10 or 30 mg/kg. Thus, the present results indicate that Scutellariae Radix, Glycyrrhizae Radix, Ginseng radix and Coptidis Rhizoma-derived components are involved in the antidiarrhoeal action of TJ-14. PMID- 10479756 TI - Antiplasmodial activity of selected sudanese medicinal plants with emphasis on Acacia nilotica. AB - Twenty-two plant organs from eleven plants comprising five families were extracted and screened for antiplasmodial activity in vitro against Plasmodium falciparum 3D7 (chloroquine sensitive) and Dd2 (chloroquine resistant and pyrimethamine sensitive). Fifty nine percent of plant extracts from 22 extracts exerted activity on P. falciparum strain 3D7 with an IC(50) less than 50 microg/mL, whereas 43% of plant extracts showed an IC(50) value within 50 microg/mL on Dd2 strains. Plant extracts from Gardenia lutea, Haplophyllum tuberculatum, Cassia tora, Acacia nilotica and Aristolochia bracteolata possessed IC(50) values less than 5 microg/mL on both tested strains. Bioassay guided fractionation of A. nilotica revealed that the ethyl acetate extract possessed the highest activity (IC(50) = 1.5 microg/mL). Fraction 2 (R(f) = 0.75) prepared by preparative chromatography showed the highest activity on P. falciparum (IC(50) = 1.7 microg/mL). Phytochemical analysis indicated that the most active phase contained terpenoids and tannins and was devoid of alkaloids and saponins. The effect of plant extracts on lymphocyte proliferation showed low toxicity to the human cells. This plant has been subjected to long term clinical trials in folk medicine and is a promising plant. PMID- 10479757 TI - The antioxidative effect of ganhuangenin against lipid peroxidation. AB - The antioxidative effect of ganhuangenin (GHG), isolated from Scutellaria baicalensis Georgi, was examined by measuring its ability to suppress the formation of phospatidylcholine hydroperoxide (PCOOH). The results show that a pretreatment with GHG effectively suppressed PCOOH formation, which was initiated by the peroxyl-generating oxidant, AAPH (2,2'-azobis-2-aminopropane hydrochloride). The protective action of GHG against the formation of the PCOOH was observed in liver, lung, and kidney. When compared with other known antioxidants, we found the antioxidative potency of GHG to be greater than that of alpha-tocopherol. Our data strongly indicate that GHG is a powerful antioxidant against lipid peroxidation and is, therefore, responsible for this prophylactic effect. PMID- 10479758 TI - The antihyperglycaemic blend effect of traditional chinese medicine byakko-ka ninjin-to on alloxan and diabetic KK-CA(y) mice. AB - The antihyperglycaemic interaction (blend effect) of component crude drugs included in a traditional Chinese prescription, Byakko-ka-ninjin-to (BN; consisted of five crude drugs, ginseng, anemarrhena,-licorice, gypsum and rice) was investigated using genetically obese diabetic KK-CA(y) mice and alloxan diabetic mice. The water extract of ginseng, anemarrhena, licorice and gypsum when individually tested markedly lowered blood glucose levels in diabetic animals. Antihyperglycaemic effects of ginseng-anemarrhena and ginseng-licorice combinations were decreased compared with the respective individual-effects. Such decreased effects were partially reversed by adding gypsum or by substituting CaCl(2) for gypsum. The combination of D-O-ANa and glycyrrhizin, respectively the main constituents from ginseng and licorice, also demonstrated a decreased effect compared with the respective individual effects. The CaCl(2) at 0.5 mg/kg (i.p.) was sufficient to partially reverse these effects. These results indicate that antihyperglycaemic effects of BN depend on four crude drugs and Ca(2+) in the blend. PMID- 10479759 TI - Separation and some properties of Aloe vera L. leaf pulp lectins. AB - The separation and partial purification of two lectins from the leaf pulp of Aloe vera L. (=barbadensis Miller) is presented. The fraction showing haemagglutinating activity was precipitated at 50% ammonium sulphate concentration from the crude leaf pulp extract. The precipitate thus obtained, after dialysis, was applied to a hydroxylapatite column. Stepwise elution resulted in two peaks showing haemagglutinating activity eluted with 5 mM (Aloctin I) and 20 mM (Aloctin II) phosphate buffers. Haemagglutinating activity was estimated visually by adding a 4% rabbit erythrocyte suspension to serial two fold dilutions of the lectins in microtitration plates. None of the 20 sugars tested inhibited haemag--glutinating activity of Aloctin I up a concentration of 500 mM. Aloctin II was inhibited by N-acetyl-D--galactosamine at 250 mM concentration. Of 10 metal ions tested, only Al(3+) salts were found to activate Aloctin I and II. On the other hand, it was shown that neither lectin possessed any alpha- and beta- galactosidase or alpha- and beta- glucosidase activity. The lectins were of glycoprotein structure containing approximately 5% neutral sugar. The specificity of the lectins towards human and rat erythrocytes was investigated. PMID- 10479760 TI - Antimicrobial potential of Spondias mombin, Croton zambesicus and Zygotritonia crocea. AB - The antimicrobial activity of Spondias mombin, Croton zambesicus and Zygotritonia crocea sourced from herbalists through an ethnobotanical survey of Egbado-South in Ogun State in Nigeria is reported. The extracts of the leaf of Spondias mombin and the stem bark of Croton zambesicus exhibited wide spectrum antibacterial effects comparable to those of ampicillin and gentamycin. The antifungal effect of extracts of Zygotritonia crocea and Croton zambesicus were comparable to those of tioconazole. There is justification for the use of these little studied medicinal plants as antiinfective agents in traditional medicine. PMID- 10479761 TI - Bioassay of phytoestrogen in herbal medicine used for postmenopausal disorder using transformed MCF-7 cells. AB - It is well known that plants contain substances with oestrogen activity comparable to that in animals. In animals, oestrogens are steroid hormones while plants contain oestrogen with various chemical characteristics, referred to as phytoestrogens. These especially, may be responsible for part of the effects of Chinese herbal prescriptions for postmenopausal disorders in which oestrogen deficiency is considered the primary cause. In addition, the presence or absence of oestrogen activity in herbal medicine is considered extremely important for patients in whom oestrogen administration is contraindicated. The most important action of oestrogen is the transcriptional regulation of specific genes via oestrogen receptors (ER). Therefore, we have established a highly sensitive bioassay system by placing oestrogen responsive elements (ERE) upstream to the reporter gene, and have used this assay to determine the oestrogen activity in herbal medicine. As a result, phytoestrogens were found in many Chinese herbal prescriptions for postmenopausal disorder, and some prescriptions were considered to effectively administer hormone replacement therapy (HRT). However, since oestrogen activities in most Chinese herbal prescriptions were thought to be weak, it was considered that these prescriptions improved postmenopausal disorder based on a mechanism differing from that of oestrogen replacement therapy. PMID- 10479762 TI - Antihyperglycaemic effect of Mangifera indica in rat. AB - The leaves of Mangifera indica are used as an antidiabetic agent in Nigerian folk medicine. To determine whether or not there is a scientific basis for this use, the effect of the aqueous extract of the leaves on blood glucose level was assessed in normoglycaemic, glucose - induced hyperglycaemic and streptozotocin (STZ)-induced diabetic rats. The aqueous extract given orally (1 g/kg) did not alter the blood glucose levels in either normoglycaemic or STZ-induced diabetic rats. In glucose - induced hyperglycaemia, however, antidiabetic activity was seen when the extract and glucose were administered simultaneously and also when the extract was given to the rats 60 min before the glucose. The hypoglycaemic effect of the aqueous extract was compared with that of an oral dose of chlorpropamide (200 mg/kg) under the same conditions. The results of this study indicate that the aqueous extract of the leaves of Mangifera indica possess hypoglycaemic activity. This action may be due to an intestinal reduction of the absorption of glucose. However, other different mechanisms of action cannot be excluded. PMID- 10479763 TI - Hepatoprotective activity of a herbal protein CI-1, purified from Cajanus indicus against beta-galactosamine HCl toxicity in isolated rat hepatocytes. AB - A herbal protein, CI-1 purified from a leguminous plant Cajanus indicus, showed dose dependent-(1.5-6.0 mg/kg x 7 days) protective activity on isolated hepatocytes (ex vivo) against beta-galactosamine-HCl induced hepatic damage in rats. It enhanced the percent viability of the hepatocytes following beta- galactosamine treatment. CI-1 was also effective in counteracting the toxic effects of beta-galactosamine, as shown by reversed levels of the altered enzymes, glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transminase (GPT) and alkaline phosphatase (ALP) both in isolated hepatic cells as well as in serum. In comparison silymarin, a known hepatoprotective agent, produced dose related protection at-relatively much higher doses than CI-1. PMID- 10479764 TI - Protection against cytotoxic effects of arsenic by dietary supplementation with crude extract of Emblica officinalis fruit. AB - Dietary administration of a crude aqueous extract of Emblica officinalis Gaertn. fruit reduced significantly the cytotoxic effects of sodium arsenite administered orally. The crude extract (685 mg/kg bw) was given daily by gavaging to age and sex matched laboratory bread Swiss albino mice for 7 and 14 days, followed by a single dose of sodium arsenite (2.5 mg/kg bw = 1/10 of LD(50)). The animals were killed after 24 h and chromosome preparations made following a schedule of colchicine-fixative-air drying-Giemsa. The endpoints screened were chromosomal aberrations and damaged cells. The crude extract reduced arsenic damage bringing the cells almost to the normal level. PMID- 10479765 TI - Modulatory effects of ixora coccinea flower on cyclophosphamide-induced toxicity in mice. AB - The active fraction of Ixora coccinea flowers showed chemoprotective effects on cyclophosphamide - induced toxicity by increasing the life span of treated mice, preventing body weight loss and maintaining near normal leucocyte and haemoglobin levels compared with cyclophosphamide treated controls. Decreased serum glutamate pyruvate transaminase (SGPT) and serum alkaline phosphatase (SAKP) levels in the Ixora coccinea treated groups indicated protection against hepatic toxicity. The active fraction was identified by phytochemical methods as the triterpenoid, ursolic acid. PMID- 10479766 TI - Screening method for PAF antagonist substances: on the phenolic compounds from Impatients balsamina L. AB - A blood pressure monitoring system was developed for studying antagonist effects against platelet activating factor (PAF), a chemical mediator of anaphylactic hypotension. Using this system, we were able to determine the inhibitory effects of a 35% ethanol extract (IB) from the petals of Impatiens balsamina L. against PAF-induced hypotension and the active principal compounds from IB. One mechanism of the antianaphylactic effects of IB was characterized as PAF-antagonist effects. PMID- 10479767 TI - Effect of extract prepared from the roots of Paeonia lactiflora on endothelium dependent relaxation and antioxidant enzyme activity in rats administered high fat diet. AB - This study examined the effect of Paeoniae Radix (PR) on endothelial function and the activity of superoxide dismutase (SOD) of erythrocytes in rats administered a high-fat diet. Administration of the extract of PR increased the endothelium dependent relaxation and the activities of SOD compared with high the cholesterol diet group significantly. Hypercholesterolaemia induced an increase of endothelial superoxide anion and endothelial dysfunction. Paeoniae Radix is suggested to have a protective effect on endothelial cells and their function. PMID- 10479768 TI - Triterpene antioxidants from ganoderma lucidum. AB - Ganoderma lucidum was studied for its antioxidative activity by bioassay guided isolation in conjunction with in vitro tests. The powdered crude drug was treated with boiling water and the aqueous extract (Ex1) was further separated to obtain terpene and polysaccharide fractions. The two fractions and Ex1 were screened for their antioxidative effect against pyrogallol induced erythrocyte membrane oxidation and Fe (II)-ascorbic acid induced lipid peroxidation. All tested samples showed antioxidative activities in a dose dependent manner and the terpene fraction was found to possess the highest effect compared with the others. Chemical isolation of the terpene fraction resulted in the detection of ganoderic acids A, B, C and D, lucidenic acid B and ganodermanontriol as major ingredients. PMID- 10479769 TI - Toxicity of neem seed oil (Azadiracta indica) against the larvae of amblyomma variegatum a three-host tick in cattle. AB - The in vitro toxicity of neem seed oil (Azadiracta indica, family: Meliaceae, 'Dogon yaro' in Hausa -language) was tested against the larvae of a three-host tick, Amblyomma variegatum (family: Ixodidae or hard tick) parasitic to cattle commonly found in Nigeria. Undiluted neem oil (100% concentration) was found to kill all (100% mortality) the larvae after 48 h. The toxicity was concentration and time dependent. PMID- 10479770 TI - Activities of silymarin and its flavonolignans upon low density lipoprotein oxidizability in vitro. AB - Silymarin, a standardized extract from Silybum marianum, inhibited in vitro the copper-induced oxidation of human LDL in a concentration-dependent manner. Silybin, a main flavonolignan of silymarin, appeared to be responsible for this LDL antioxidant effect. Silychristin and silydianin, other flavonolignans of silymarin, acted rather as pro-oxidants, but with regard to their content in silymarin, it did not contribute significantly to the reduction of the total LDL antioxidant capacity of silymarin. PMID- 10479771 TI - In vitro antitrypanosomal activity of Moringa stenopetala leaves and roots. AB - The leaves and the root extracts of Moringa stenopetala were tested in vitro against trypomastigotes of Trypanosoma brucei, Trypanosoma cruzi and L. donovani amastigotes. The fresh root wood ethanol extract and the dried leaves acetone extract were found to be active against T. brucei with an ED(50) value of 9.2 microg/mL and 10.0 microg/mL respectively. All the other extracts were inactive against all the tested parasite forms. PMID- 10479772 TI - Studies on the mode of action of the essential oil of lavender (Lavandula angustifolia P. Miller). AB - Lavender (Lavandula angustifolia, P. Miller) is used in aromatherapy as a holistic relaxant and is said to have carminative, antiflatulence and anticolic properties. Its sedative nature, on inhalation, has been shown both in animals and man. Lavender has a spasmolytic activity on guineapig ileum and rat uterus in vitro and it also decreases the tone in the skeletal muscle preparation of the phrenic nerve-diaphragm of rats. As the mechanism of action has not been studied previously, the spasmolytic activity was studied in vitro using a guinea-pig ileum smooth muscle preparation. The mechanism of action was postsynaptic and not atropine-like. The spasmolytic effect of lavender oil was most likely to be mediated through cAMP, and not through cGMP. The mode of action of linalool, one of lavender's major components, reflected that of the whole oil. The mode of action of lavender oil resembled that of geranium and peppermint oils. PMID- 10479773 TI - Patents alert PMID- 10479774 TI - Selected bibliography PMID- 10479775 TI - Defending vaccines from the enemy within. PMID- 10479776 TI - The selectivity of action of an antiherpetic agent, 9-(2-hydroxyethoxymethyl) guanine. Reproduced from Proc. Natl. Acad. Sci. USA 74, 5716-5720 (1977) PMID- 10479777 TI - The discovery of two new divergent STLVs has implications for the evolution and epidemiology of HTLVs. AB - We have isolated and characterised two divergent simian T-lymphotropic viruses (STLV), not belonging to the established human and simian T-lymphotropic virus lineages HTLV-1/STLV-1 and HTLV-2. STLV-L, from an Eritrean sacred baboon (Papio hamadryas), has been typed as a third type of simian T-lymphotropic virus, distinct from HTLV-1/STLV-1 and HTLV-2. The other virus, isolated from Congolese bonobos (Pan paniscus), is a distinct member of the HTLV-2 clade and has been designated STLV-2. The isolation of these two simian viruses shows that the spectrum of HTLVs/STLVs is larger than previously expected. Our data indicate that the two lineages STLV-L and HTLV-2/STLV-2 are of African origin, while the HTLV-1/STLV-1 lineage has been shown to be of Asian origin. These data, together with our phylogenetic analyses, suggest an African origin of the HTLV/STLV ancestor, which provides new clues about virus dissemination. Furthermore, the atypical serological profiles exhibited by STLV-L or STLV-2 infected animals in western blot, raise questions about the efficiency of current screening methods to type highly divergent HTLVs/STLVs. Considering the growing interest in xenotransplantations, more epidemiological and biological knowledge of simian and human T-lymphotropic viruses is necessary to estimate the risk of interspecies transmissions. PMID- 10479778 TI - Mechanism and application of genetic recombination in herpesviruses. AB - Herpes simplex virus type 1 (HSV-1) is a ubiquitous human pathogen that latently infects sensory ganglia and encodes over 80 genes in a 152 kbp DNA genome. This well characterised virus provides a model for analysing genetic recombination in herpesviruses, a fundamental biological process by which new combinations of genetic materials are generated. The frequency of homologous recombination was estimated to be 0.0048-0.007 (0.48%-0.7%)/kb of the HSV-1 genome, determined using physical markers. The double-strand break repair model, the current model of homologous recombination, adequately explains L-S inversion of herpesvirus genomes and the recombinogenicity of the a sequence. Several herpesvirus genomes, including HSV-1 consist of a unique sequence bracketed by a pair of inverted repeat sequences. This arrangement is attributed to illegitimate recombination between molecules arranged in an inverse orientation. Junctions of unique and repeated sequences that correspond to the crossover site of illegitimate recombination are recombinogenic. Recombination is important for virus evolution, construction of mutated virus, gene therapy and vaccination in which the potential for recombination between engineered input virus and wild type virus has to be considered. PMID- 10479779 TI - Natural and iatrogenic variation in hepatitis B virus. AB - The existence of HBV as quasispecies is thought to be favoured by the infidelity of HBV RT, which would account for the emergence of the many natural mutants with point substitutions. RT infidelity may also underlie the hypermutation phenomenon. Indeed, the oft-reported point mutation in the preC gene that leads to failure of HBeAg synthesis may be driven by a hypermutation-related mechanism. The presence of mutants with deletions and insertions involving single nucleotides and oligonucleotides at specific positions in the genome, and of mutants with deletions of even longer stretches particularly in the C gene, suggests that other mutagenic mechanisms operate. Candidates include slippage during mispairing between template and progeny DNA strand, the action of cellular topoisomerase I, and gene splicing using alternative donor and acceptor sites. Natural substitutions, deletions or insertions involving the Cp/ENII locus in the X gene can significantly alter the extent of viral replicative activity. Similar mutations occurring at other locations of Cp/ENII, and at B-cell epitope sites of the S gene are associated with failure to detect serological markers of HBV infection. HBV variation can also arise from recombination between coinfecting strains. S gene mutations that become evident following HBIG administration and HBV vaccination are all point substitutions, as are mutations in functional RT domains of the P gene after treatment with viral RT-inhibitory drugs. Widespread and long-term use of prophylactic and therapeutic agents may potentially generate serologically occult HBV variants that might become difficult to eradicate. PMID- 10479780 TI - European CPMP gives positive opinion on lamivudine for chronic hepatitis B. PMID- 10479781 TI - These magic moments PMID- 10479782 TI - Effects of testing context on ball skill performance in 5-year-old children with and without developmental delay. AB - BACKGROUND AND PURPOSE: Physical therapists often test ball-playing skills of children with disabilities using standardized testing, which may not predict performance of ball skills in games with peers. This type of testing is used by physical therapists to determine whether children have delays in ball-handling skills. The purpose of this study was to compare ball skill performance of children with and without developmental delay in a one-to-one testing situation and in a structured game with peers. SUBJECTS: Five-year-old children with developmental delay (n=20) and 5-year-old children without disabilities (n=20) participated in the study. METHODS: We used the Peabody Developmental Motor Scales receipt and propulsion scale to test children one-to-one with a therapist and during a structured game with peers. RESULTS: Subjects without developmental delay performed better than subjects with developmental delay under both testing conditions. Children with developmental delay performed better in the one-to-one testing condition than in the game with peers. The performance of children without developmental delay did not differ under the 2 conditions. Boys performed better than girls. CONCLUSION AND DISCUSSION: Physical therapists should consider the potential effect of environment on the ball-handling skills of children with disabilities. PMID- 10479783 TI - Influence of age on length and passive elastic stiffness characteristics of the calf muscle-tendon unit of women. AB - BACKGROUND AND PURPOSE: Therapeutic stretching of the calf muscle-tendon unit is used to increase its length and to ameliorate decreased dorsiflexion range of motion (ROM), but the influence of age on the passive properties of the calf muscle-tendon unit has not been studied adequately The purpose of this study was to examine the influence of age on length and passive elastic stiffness (PES) characteristics of the calf muscle-tendon unit when stretched through the full, available dorsiflexion ROM. SUBJECTS: Twenty-four younger women (aged 20-39 years), 24 middle-aged women (aged 40-59 years), and 33 older women (aged 60-84 years) participated. METHODS: An isokinetic dynamometer was used to passively stretch the right calf muscle-tendon unit from relaxed plantar flexion to the maximal angle of available dorsiflexion at 5(us-1. The maximal passive resistive torque was measured, and passive angle-torque curves were constructed for a full ROM from an initial angle of passive resistive torque to the maximal dorsiflexion angle. The full ROM represented length extensibility. The average PES was calculated for this full stretch ROM and for the first half and the last half of this stretch ROM. The maximal passive dorsiflexion angle, maximal passive resistive torque, angular change for the full stretch ROM, and average PES for the full stretch ROM and the first half and the last half of the full stretch ROM were examined for group differences and their relationships with age. RESULTS: The maximal passive dorsiflexion angle, maximal passive resistive torque, angular change for the full stretch ROM, and average PES within the last half of the full stretch ROM were less for the older women than for the younger women. Age was negatively associated with these variables. CONCLUSION AND DISCUSSION: Decreased maximal passive dorsiflexion ROM in older women was associated with decreased maximal passive resistive torque, decreased calf muscle-tendon unit length extensibility, and decreased average PES within the last half of their available passive dorsiflexion ROM. PMID- 10479784 TI - Bactericidal effect of 0.95-mW helium-neon and 5-mW indium-gallium-aluminum phosphate laser irradiation at exposure times of 30, 60, and 120 seconds on photosensitized Staphylococcus aureus and Pseudomonas aeruginosa in vitro. AB - BACKGROUND AND PURPOSE: Studies have demonstrated a bactericidal effect of laser irradiation when lasers with power outputs of (6 mW are directed toward pathogenic or opportunistic bacteria previously treated with a photosensitizing agent. The purpose of this study was to determine the bactericidal capabilities of irradiation from lasers with power outputs of less than 6 mW on photosensitized microorganisms. METHODS: Two bacteria that commonly infect skin lesions, Staphylococcus aureus and Pseudomonas aeruginosa, were used. The 2 lasers used, the 0.95-mW helium -neon laser and the 5-mW indium-gallium-aluminum phosphate laser, emit light at a wavelength close to the absorption maxima of the sensitizing agent chosen, toluidine blue O. This agent was used because of its proven effectiveness in sensitizing bacteria. For each bacterial strain, toluidine blue O was added to a 108 cells/mL solution until a 0.01% weight/volume ratio was obtained. These mixtures were spread on agar-coated petri dishes, which were then exposed to 1 of the 2 lasers for 30, 60, and 120 seconds. The cultures were then grown overnight and examined for one or more visible zones of inhibition. The areas surrounding the irradiated zone provided a control for the effects of toluidine blue O alone. To determine the effects of laser irradiation without prior toluidine blue O sensitization, separate plates were established using unsensitized bacteria. RESULTS: Although inconsistencies between plates were noted, both lasers produced at least one zone of inhibition in both bacterial species at all 3 time periods. The 5-mW laser, however, produced a greater number of these zones. CONCLUSION AND DISCUSSION: Laser-induced microbial killing of photosensitized organisms could have clinical applications in the treatment of infected skin lesions, pending in vivo studies. PMID- 10479785 TI - An application of upper-extremity constraint-induced movement therapy in a patient with subacute stroke. AB - BACKGROUND AND PURPOSE: The purpose of this case report is to demonstrate the application of constraint-induced movement therapy with an individual with upper extremity hemiparesis within 4 months after sustaining a cerebrovascular accident (stroke). Such patients often fail to develop full potential use of their affected upper extremity, perhaps due to a "learned nonuse phenomenon." CASE DESCRIPTION: The patient was a 61-year-old woman with right-sided hemiparesis resulting from an ischemic lacunar infarct in the posterior limb of the left internal capsule. The patient's less-involved hand was constrained in a mitten so that she could not use the hand during waking hours, except for bathing and toileting. On each weekday of the 14-day intervention period, the patient spent 6 hours being supervised while performing tasks using the paretic upper extremity. Pretreatment, posttreatment, and 3-month follow-up outcome measures included the Wolf Motor Function Test and the Motor Activity Log (MAL). OUTCOMES: For the Wolf Motor Function Test, both the mean and median times to complete 16 tasks improved from pretreatment to posttreatment and from posttreatment to follow-up. Results of the MAL indicated an improved self-report of both "how well" and "how much" the patient used her affected limb in 30 specified daily tasks. These improvements persisted to the follow-up. DISCUSSION: Two weeks of constraining the unaffected limb, coupled with practice of functional movements of the impaired limb, may be an effective method for restoring motor function within a few months after cerebral insult. Encouraging improvements such as these strongly suggest the need for a group design that would explore this type of intervention in more detail. PMID- 10479786 TI - Anatomy and biomechanics of the first ray. PMID- 10479787 TI - [Anticoagulant therapy can be safer. An active follow-up and written care programs reduce the number of complications]. PMID- 10479788 TI - [Identification of cancer genes has made the testing of predisposition possible. Genetic counseling and mutation screening can result in new therapeutic methods]. PMID- 10479789 TI - [Good physician-patient relation can prevent disinformation]. PMID- 10479790 TI - [Management of bee and wasp stings!]. PMID- 10479791 TI - [Time for a national consensus on the diagnosis of peptic ulcer]. PMID- 10479792 TI - [Open discussion on the choice of drugs is better than centralized supervision]. PMID- 10479793 TI - [It should not be necessary for elderly patients with fractures to wait for surgery]. PMID- 10479794 TI - [Misleading information about implantable defibrillators]. PMID- 10479795 TI - [Transplanted nerve cells survive and are functional for many years]. AB - During the past decade neuronal grafting has been explored as a novel approach to the treatment of neurodegenerative disease. About 250 patients with advanced Parkinson's disease and 20-30 patients with Huntington's disease have received embryonic neuronal grafts. The results have shown such grafts to survive and function for many years in the diseased brain, and to induce significant and lasting therapeutic effects. The article gives a review of experience in current trials, discussed in the light of the practical and ethical problems that need to be solved in order to develop the neuronal graft technique as a useful and generally acceptable form of therapy. PMID- 10479796 TI - [Further clarification of functional issues in cystic fibrosis. Current research and future prospects]. AB - First rate collaboration between clinicians and research scientists in a multiplicity of fields have brought new hope to patients with cystic fibrosis (CF). The gene, mutations of which give rise to the disease, has been exhaustively mapped, and the functional defects are becoming steadily clearer. Diagnosis is continually being improved and simplified. Neonatal screening has been introduced in many countries and has yielded good results. Promising new advances in treatment include inhalatory DNase (deoxyribonuclease), lung and liver transplantation, UDCA (ursodeoxycholic acid) against cirrhosis, and in vitro fertilisation for men with CF. Pseudomonas species are being combatted more and more effectively with new antibiotics, with immunoglobulins (IgY) for prophylaxis, and possibly new vaccines to come. Future treatment strategies, designed to correct anomalies of cellular biology, are already undergoing clinical trials, and gene therapy using a variety of vectors is undergoing phase 1 trials. A definitive cure remains a realistic hope. PMID- 10479797 TI - [Survey of complications following treatment with anticoagulants. A computerized search for hemorrhagic complications completes manual reporting]. PMID- 10479798 TI - [Is home care really more expensive? A report on costs of care is questioned]. PMID- 10479799 TI - [A harvest time for genomic research]. AB - The article consists in a review of the human genome project, launched a decade ago to characterise the entire human genome. The project has proved highly successful, due both to the economy of so large scale an endeavour and to the value of gaining access to such an abundance of biological information. Accordingly, similar approaches have also been adopted in efforts to characterise the entire range of genes expressed as mRNA and as protein. Genomic information has become an invaluable asset to biomedical research, and both the information obtained and the methodology developed are now important adjuncts of pharmaceutical research. Applications in clinical medicine follow, albeit at a slower rate. PMID- 10479800 TI - [Being an intern on call when a child is born in the 22nd week of pregnancy]. PMID- 10479802 TI - [Mites behind an "island rash" during black currant picking on an island in the gulf of Gavle]. PMID- 10479801 TI - [A comment: in the borderland between a late abortion and premature delivery]. PMID- 10479803 TI - "Point four" the future. PMID- 10479804 TI - Should there be laws mandating domestic violence reporting? PMID- 10479805 TI - FOCUS on preparation for RSV season. AB - The purpose of this article is to describe one institution's successful program to deal with the many challenges incurred by a pediatric service during the Respiratory Syncytial Virus (RSV) season. RSV is a winter phenomenon that appears each year in communities across the globe and results in up to a 60% increase in admissions to children's hospitals and pediatric units. This increase places a huge demand on healthcare institutions, for the census and intensity of illness requires additional qualified pediatric nurses, physicians, and respiratory therapists as well as additional bed space and equipment. This institution developed strategies to cope with RSV, including using work force teams and the continuous quality improvement (CQI) process. PMID- 10479806 TI - Psychoneuroimmunology and preterm birth. A holistic model for obstetrical nursing practice and research. AB - This article presents a framework through which nurses can conceptualize premature labor and birth for both practice and research. Use of the psychoneuroimmunology (PNI) model may guide the study of the problem of preterm birth in a more holistic manner, discovering relationships between the body and the mind that may affect how nurses can intervene to prevent premature birth. Nursing assessment of risk needs to include those situations that may lead to increased stress or anxiety, as is supported by research based on the PNI model. Reduction of stressors that lead to physiological changes related to the stress response can affect the incidence of preterm labor. Interventions to decrease stress and poor coping behaviors need to be tested and integrated into practice. PMID- 10479807 TI - Therapeutic relationships. Evolution of the Children's Hospital of Philadelphia model. AB - The evolution of family-centered pediatric care coupled with the increasing complexity of healthcare environments has had a tremendous impact on the nature of relationships between nursing staff and the children and families they serve. In order to address this, in 1988 the Nursing Department at the Children's Hospital of Philadelphia (CHOP) identified parameters for the development and maintenance of therapeutic relationships. A therapeutic relationship is an interactive relationship with a child and family that is caring, clear, boundaried, positive, and professional. It encompasses the philosophy of the institution, empowerment of the caregivers, and empowerment for families. Utilizing family-centered care as a framework for the delivery of care, this article describes the evolution of therapeutic relationships as a standard of practice including the articulation of principles, clinical examples, implications for practice, and strategies for assessment, management, and evaluation. PMID- 10479808 TI - Kangaroo Care for a restless infant with gastric reflux. One nurse midwife's personal experience. PMID- 10479809 TI - Perspectives of women whose sons become adolescent fathers. AB - PURPOSE: The purpose of this study was to describe adolescent fatherhood from the perspectives of these young men's mothers. DESIGN: A descriptive qualitative design was chosen because of the exploratory nature of this study. METHODS: Data collection and analysis were guided by naturalistic inquiry that permits theory to emerge from the data. The data were collected during a 1-hour focus group interview performed at a public high school located proximal to a large public housing project in a large Midwestern city. Five mothers of adolescent fathers were recruited from a variety of sources using a modified snowball approach. RESULTS: The seven major themes that emerged from the data were: barriers to fatherhood, value of fatherhood, transition to fatherhood, competencies of fatherhood, role-set relationships, social norms of fatherhood, and father-child contact. These women indicated that they felt their sons' youth influenced every aspect of the fatherhood experiences. CLINICAL IMPLICATIONS: Nurses who work with adolescent parents are ideally situated to encourage the support of adolescent fathers and their families. Future research should identify methods useful for supporting or discouraging paternal involvement of adolescent fathers, and effective intervention strategies for adolescent fathers and their families. PMID- 10479810 TI - Assessment of stature in children with orofacial clefting. AB - PURPOSE: To assess stature in children with orofacial clefting to determine whether this population is at risk for short stature and if growth failure is caused by hypothalamic-pituitary dysfunction or if it is related to age, sex, or type of cleft. DESIGN: Cross-sectional. METHODS: All children 3 to 12 years old with nonsyndromic orofacial clefts who were evaluated as outpatients were measured; those who demonstrated growth failure were to have an evaluation of their hypothalamic-pituitary function. Data were also collected on sex, age, and type of cleft. RESULTS: Children with orofacial clefting had significantly more growth failure than the general population (p < 0.005). The parents of four of the five children with growth failure refused further evaluation because "no one else was ever concerned about my child's height." CLINICAL IMPLICATIONS: The high rate of growth failure in this population emphasizes the need for precise growth assessment of children with orofacial clefting. Growth failure should not be ignored or minimized in populations of children having other significant health care problems. PMID- 10479811 TI - Grab their attention! Make your point! AB - Improving the education we offer is an important goal for nurses. Advances in computer presentation software have given us the ability to use visuals and sound to enhance learning, whether for patient education or staff development. Better visual design of your presentation should achieve four basic goals in communicating your message: (1) ensure legibility, (2) reduce the effort required to interpret the message, (3) increase the viewer's active engagement with the message, and (4) focus attention on the most important parts of the message. Guidelines are given in this article to meet each of these goals. PMID- 10479812 TI - Legal issues in the treatment of infertility. PMID- 10479813 TI - Prenatal instruments. PMID- 10479815 TI - ADHD. PMID- 10479814 TI - Fetal heart rate monitoring certification. PMID- 10479816 TI - The time-course of negative priming: little evidence for episodic trace retrieval. AB - An episodic trace retrieval (ETR) explanation of negative priming (NP) predicts that the NP effect should be sensitive to the timing of delays between trials (Neill & Valdes, 1992; Neill, Valdes, Terry, & Gorfein, 1992). Specifically, according to ETR, (1) NP is affected by the response-stimulus interval (RSI) before the prime display, and (2) NP decays when RSI is manipulated within groups but not when RSI is manipulated between groups. Two localization tasks and two identification tasks are reported that question the reliability of these findings. The results suggest that there is little in the time-course literature that uniquely supports the ETR theory of NP. Instead, the results seem more compatible with either a dual-mechanism account (Kane, May, Hasher, Rahhal, & Stoltzfus, 1997) or an integrative approach that incorporates both memory and attention processes (Milliken, Joordens, Merikle, & Seiffert, 1998). PMID- 10479817 TI - Suppression of reflexive saccades in younger and older adults: age comparisons on an antisaccade task. AB - Inhibitory control of prepotent responses has been examined by using the antisaccade task, during which a reflexive saccade toward a peripheral onset must be suppressed before an eye movement in the opposite direction from the onset can be executed. In the present experiments, we sought to determine whether older and younger adults would perform similarly on this task. Older adults had a harder time suppressing their reflexive responses, as measured by an increase in the proportion of saccade direction errors. Despite an age-related decline in saccade direction accuracy, the increase in saccade latency associated with the antisaccade condition was the same for both younger and older adults. These results support the view that the effectiveness of inhibitory control declines with age (Hasher & Zacks, 1988; Hasher, Zacks, & May, 1999). PMID- 10479818 TI - Age of acquisition, word frequency, and the role of phonology in the lexical decision task. AB - In five experiments, we examined the respective roles of word age of acquisition (AoA) and frequency in the lexical decision task. The two variables were manipulated orthogonally (while controlling for concreteness and length) in fully factorial designs. Experiment 1 was a conventional lexical decision task, and Experiments 2-5 involved various attempts to interfere with reliance upon phonology. In Experiment 2, only orthographically illegal nonwords were used; in Experiment 3, pseudohomophone nonwords; in Experiment 4, articulatory suppression by the recitation of a nursery rhyme; and in Experiment 5, articulatory suppression by the repetition of a single word. The same basic pattern of results was observed in all experiments: There were main effects of both AoA and frequency, which interacted in such a way that the AoA effect was larger for low- than for high-frequency words. Although the AoA effect was reduced by manipulations intended to interfere with phonological processing, the manipulations did not eliminate the effect. The results are discussed in terms of current models of reading in which it is proposed that AoA has its primary effect on the retrieval of lexical phonology, which appears to be consulted automatically in the lexical decision task. PMID- 10479819 TI - Recency and lexical preferences in Spanish. AB - One experiment provided evidence in support of Gibson, Pearlmutter, Canseco Gonzalez, and Hickok's (1996) claim that a recency preference applies to Spanish relative clause attachments, contrary to the claim made by Cuetos and Mitchell (1988). Spanish speakers read stimuli involving either two or three potential attachment sites in which the same lexical content of the two-site conditions appeared in a different structural configuration in the three-site conditions. High attachment was easier than low attachment when only two sites were present, but low attachment was preferred over high attachment, which was in turn preferred over middle attachment, when three sites were present. The experiment replicated earlier results and showed that (1) attachment preferences are determined in part by a preference to attach recently/low, and (2) lexical biases are insufficient to explain attachment preferences. PMID- 10479820 TI - Eponymous verb phrases and ambiguity resolution. AB - To understand eponymous verb phrases such as "do a John Travolta," readers cannot merely select a sense out of a mental lexicon (sense selection). They must create new senses (sense creation) by retrieving salient information from memory. We conducted two experiments to test the hypothesis that these processes of memory retrieval parallel those used for ordinary lexical ambiguities. To prepare for Experiment 1, we gathered readers' interpretations of eponymous verb phrases like "do a John Travolta" to establish dominant and subordinate interpretations. We then wrote story contexts that biased comprehension toward one or the other interpretation. In Experiment 1, paraphrase judgment times were used to demonstrate that dominant meanings are privileged in the sense that they are accessible even when the story creates a subordinate bias. In Experiment 2, this privilege faded somewhat when there was a delay before the paraphrase judgment. We discuss the results with respect to the distinction between sense selection and sense creation. PMID- 10479821 TI - Acquisition of novel traces in short-term implicit memory: priming for nonwords and new associations. AB - McKone (1995) reported a short-lived repetition priming effect (up to 8 sec and three intervening items), superimposed on long-term priming. In lexical decision and naming, decay of this short-term implicit memory was faster for pseudowords than for words, suggesting an explanation in terms of transient activation of preexisting lexical representations. Here, we present two cases where, in contrast, preexperimental familiarity did not affect short-term priming, indicating acquisition of novel traces. Experiment 1 determined repetition priming in same-different judgments to lowercase-uppercase pairs for words, and for nonwords with three levels of wordlikeness. Across lags of 0, 1, and 6 intervening items (2-14 sec), short-term priming was the same for all stimuli, even random letter strings. Experiments 2 and 3 assessed priming in a double lexical decision task for old associations (orange-apple) and new associations (cigar-errand). Short-term priming for the association was equal in both cases. PMID- 10479822 TI - The effects of levels-of-processing and organization on conceptual implicit memory in the category exemplar production test. AB - The distinction between item-specific and relational information (Hunt & Einstein, 1981) may be relevant to accounts of conceptual priming in the category exemplar production task. In three experiments, the implications of this hypothesis were tested by examining the effects of organization and levels-of processing (LOP) on this implicit test. Consistent with the hypothesis, the effects of LOP were greater when study lists were organized by category than when they were presented randomly. Furthermore, when subjects claiming test awareness or intentional retrieval were excluded from the analysis, the LOP effect was reduced (and, in fact, eliminated) in the random list condition but remained robust in the categorized condition. Finally, the experimental design of the LOP manipulation (i.e., mixed-list within, blocked-list within, or between subjects) did not moderate the effects of LOP on this priming task. PMID- 10479823 TI - Monitoring item and source information: evidence for a negative generation effect in source memory. AB - Item memory and source memory were assessed in a task that simulated a social conversation. Participants generated answers to questions or read statements presented by one of three sources (faces on a computer screen). Positive generation effects were observed for item memory. That is, participants remembered topics of conversation better if they were asked questions about the topics than if they simply read statements about topics. However, a negative generation effect occurred for source memory. That is, remembering the source of some information was disrupted if participants were required to answer questions pertaining to that information. These findings support the notion that item and source memory are mediated, as least in part, by different processes during encoding. PMID- 10479824 TI - Familiarity and recollection in item and associative recognition. AB - Recognition memory for item information (single words) and associative information (word pairs) was tested immediately and after retention intervals of 30 min and 1 day (Experiment 1) and 2 days and 7 days (Experiment 2) using Tulving's (1985) remember/know response procedure. Associative recognition decisions were accompanied by more "remember" responses and less "know" responses than item recognition decisions. Overall recognition performance and the proportion of remember responses declined at similar rates for item and associative information. The pattern of results for item recognition was consistent with Donaldson's (1996) single-factor signal detection model of remember/know responses, as comparisons based on A' between overall item recognition and remember item recognition showed no significant differences. For associative recognition, however, A' for remember responses was reliably greater than for overall recognition. The results show that recollection plays a significant role in associative recognition. PMID- 10479825 TI - Effects of exact and category repetition in true and false recognition memory. AB - Two experiments used the distinction between remembering and knowing to investigate the effects of exact and category repetition in recognition memory. In Experiment 1, exact repetition enhanced remember responses but had no reliable effect on know responses. In Experiment 2, category repetition enhanced correct know responses but had no effect on correct remember responses. Category repetition also increased false positive remember and know responses. It is argued that exact repetition influences the recollection component of recognition memory via the creation of multiple episodic traces, each of which is potentially capable of supporting a remember response, whereas category repetition influences the familiarity component of recognition memory by enhancing the fluency with which test items are processed. PMID- 10479826 TI - Visual effects in picture and word categorization. AB - Two experiments are reported in which participants categorized stimuli as belonging or not belonging to the category of fruits. Blocks of pictures and words were used, with items referring to exemplars having either high or low intercategory visual similarity and/or semantic relatedness. For both pictures and words, response time was longer in the semantically related conditions than in the unrelated condition. Furthermore, there was a strong effect of visual similarity for pictures but not for words when semantic relatedness was held constant: Participants took longer to classify pictures of fruits when these were mixed with visually similar vegetables than when they were mixed with visually dissimilar vegetables. Reducing the stimulus visibility by adding a dot pattern had an additive effect for words but an interactive effect for pictures. The results are explained in terms of a unique locus for category decisions about pictures and words. PMID- 10479827 TI - The influence of prior knowledge in intentional versus incidental concept learning. AB - Previous research has demonstrated that background knowledge has a clear influence on concept learning. This influence, however, has been observed with a narrow range of intentional learning tasks. In the present experiments, the role of background knowledge was examined as a function of a variety of incidental learning tasks as well as with intentional learning tasks. The influence of prior knowledge was investigated by comparing the encoding of conceptually related co occurrences with the encoding of conceptually unrelated co-occurrences. A clear influence of prior knowledge was observed with incidental encoding, and conceptual relatedness was found to have at least as powerful an influence with incidental as with intentional learning tasks. The results indicate that many types of knowledge-based influences will not vary as a function of encoding strategy. The pervasiveness and strength of the influence of background knowledge on concept learning are discussed. PMID- 10479828 TI - The acquisition of category structure in unsupervised learning. AB - Four experiments examined the extent to which prior knowledge influences the acquisition of category structure in unsupervised learning conditions. Prior knowledge is general knowledge about a broad domain that explains why an object has the features it does. Category structure refers to the statistical regularities of features within and across categories. Subjects viewed items and then divided them up into the categories that seemed most natural. Each item had one feature that was related to prior knowledge and five features that were not. The results showed that even this small amount of prior knowledge helped subjects to discover the category structure. In addition, prior knowledge enhanced the learning of many of the category's features, and not just the features that were directly relevant to the knowledge. The results suggest that prior knowledge may help to integrate the features of a category, thereby improving the acquisition of category structure. PMID- 10479829 TI - Decision making under time pressure: an independent test of sequential sampling models. AB - Choice probability and choice response time data from a risk-taking decision making task were compared with predictions made by a sequential sampling model. The behavioral data, consistent with the model, showed that participants were less likely to take an action as risk levels increased, and that time pressure did not have a uniform effect on choice probability. Under time pressure, participants were more conservative at the lower risk levels but were more prone to take risks at the higher levels of risk. This crossover interaction reflected a reduction of the threshold within a single decision strategy rather than a switching of decision strategies. Response time data, as predicted by the model, showed that participants took more time to make decisions at the moderate risk levels and that time pressure reduced response time across all risk levels, but particularly at the those risk levels that took longer time with no pressure. Finally, response time data were used to rule out the hypothesis that time pressure effects could be explained by a fast-guess strategy. PMID- 10479830 TI - Deductive reasoning with factual, possible, and counterfactual conditionals. AB - We compared reasoners' inferences from conditionals based on possibilities in the present or the past (e.g., "If Linda had been in Dublin then Cathy would have been in Galway") with their inferences based on facts in the present or the past (e.g., "If Linda was in Dublin then Cathy was in Galway"). We propose that people construct a richer representation of conditionals that deal with possibilities rather than facts: Their models make explicit not only the suppositional case, in which Linda is in Dublin and Cathy is in Galway, but also the presupposed case, in which Linda is not in Dublin and Cathy is not in Galway. We report the results of four experiments that corroborate this model theory. The experiments show that reasoners make more inferences from conditionals based on possibilities rather than on facts when the inferences depend on the presupposed case. The results also show that reasoners generate different situations to verify and falsify conditionals based on possibilities and facts. PMID- 10479831 TI - Spatial knowledge acquisition from maps and from navigation in real and virtual environments. AB - In this study, the nature of the spatial representations of an environment acquired from maps, navigation, and virtual environments (VEs) was assessed. Participants first learned the layout of a simple desktop VE and then were tested in that environment. Then, participants learned two floors of a complex building in one of three learning conditions: from a map, from direct experience, or by traversing through a virtual rendition of the building. VE learners showed the poorest learning of the complex environment overall, and the results suggest that VE learners are particularly susceptible to disorientation after rotation. However, all the conditions showed similar levels of performance in learning the layout of landmarks on a single floor. Consistent with previous research, an alignment effect was present for map learners, suggesting that they had formed an orientation-specific representation of the environment. VE learners also showed a preferred orientation, as defined by their initial orientation when learning the environment. Learning the initial simple VE was highly predictive of learning a real environment, suggesting that similar cognitive mechanisms are involved in the two learning situations. PMID- 10479832 TI - The effect of prior experience on recognition thresholds for plane-disoriented pictures of familiar objects. AB - We investigated plane rotation effects on the minimum presentation duration that is required in order to recognize pictures of familiar objects, using the method of ascending limits. Subjects made unspeeded verification responses, selecting from 126 written alternatives. Replicating similar identification studies in which brief, masked pictures (Lawson & Jolicoeur, 1998) were presented, disorientation reduced the efficiency of recognition. Mirroring the findings in speeded picture naming studies (e.g., Jolicoeur, 1985; Jolicoeur & Milliken, 1989), but in contrast to those of Lawson and Jolicoeur (1998), orientation effects were found over a wide range of views and were attenuated but not eliminated with experience with a given object. The results bridge the findings from unspeeded verification and speeded naming tasks. They suggest that the same orientation-sensitive processes are tapped in both cases, and that practice effects on these processes are object specific. PMID- 10479833 TI - Sepsis, multi-organ dysfunction syndrome (MODS) and multiple organ failure (MOF). Prevention is better than treatment. PMID- 10479834 TI - [Total intravenous anesthesia in thoracotomy with one-lung ventilation]. AB - BACKGROUND: The aim of this study was to evaluate the benefits of total intravenous anaesthesia (TIVA), with the administration of drugs intravenously and the maintenance of pulmonary ventilation with oxygen enriched air, with respect to inhalatory anaesthesia, in thoracotomies with single lung ventilation. METHODS: The study, devoloped in collaboration between the Service of Anaesthesia and Resuscitation and the Division of Thoracic Surgery of the Hospital of Teramo, was carried out in two groups of patients chosen at random; fifty patients divided into two groups of twenty-five were studied. Pre-medication and induction was similar in both groups; maintenance, however, varied: a total intravenous anaesthesia was given to patients in group A whereas patients in group B received an inhalatory anaesthesia. These parameters were considered: pressure values, heartbeat, PaO2 and PaCO2 levels, SatO2 and EtCO2. RESULTS: Both protocols respected anaesthetic guidelines in terms of controlling pressure values, heartbeat and levels of PaCO2 during bipulmonary ventilation. Significantly statistical differences were observed in the oxygenation during one lung ventilation: the mean values of PaO2 being significantly higher in group A. CONCLUSIONS: On the basis of this experience, it can be conclude that TIVA offers the following advantages: a better oxygenation during one lung ventilation, good recovery of post operative consciousness with no psychomotor disturbances, absence of pollution in the operating theatre. PMID- 10479835 TI - [Use of remifentanil in ambulatory obstetric-gynecologic surgery. A dose-effect study]. AB - BACKGROUND: Remifentanil, a recently commercialised opioid, is characterised by a predictable and non cumulative effect which vanishes rapidly without determining side effects in the long term. These characteristics make remifentanil an ideal opioid in continuous infusion for the ambulatory surgery setting. Aim of this study was to assess the ideal dose of remifentanil, administered in bolus before propofol, in patients undergoing uterine curettage and assisted by mask ventilation in 100% oxygen. METHODS: Sixty patients, ASA status I-II, scheduled for uterine curettage, were divided into three study groups according to the bolus dose of remifentanil received before the induction agent: group A (n = 20) 1 microgram/kg; group B (n = 20) 2 micrograms/kg; group C (n = 20) 2 micrograms/kg. All patients were assisted by 100% oxygen ventilation with facial mask. During surgery the following were recorded: time to spontaneous ventilation (in case of post induction apnea); incidence of somatic and autonomic responses to surgical stress (treated with remifentanil in bolus). At the end of surgery the times to response to simple verbal commands, to discharge from the recovery room (by Aldrete score every 5') and to discharge from hospital (by PADSS score every 30') were registered. RESULTS: All patients presented post-induction apnea with a significantly more rapid return to spontaneous ventilation in group A. Six patients of group A responded to surgical stress while in groups B and C there was no need for supplementary boluses (p < 0.05). Five patients of group C were treated with atropine for bradycardia, in four of group C it was necessary to administer succinylcholine for thoracic rigidity. No significant differences regarding the anesthesia recovery times were observed. All patients were discharged from the recovery room after 10' from the end of surgery. Overall, the qualification for discharge from hospital was obtained at the second PADSS score control, except for one group A patient who incurred in metrorrhagia. CONCLUSIONS: The administration in bolus of remifentanil, before the inducing agent, permits short-term surgery in ambulatory surgery settings thanks to the rapid recovery of vital functions. Compared to the other doses, the 1.5 micrograms/kg dose guaranteed a good control over surgical stress without influencing the speed of awakening and without determining uncomfortable side effects. PMID- 10479836 TI - Clinical and hormonal response to general anaesthesia in patients affected by different degrees of mental retardation. AB - BACKGROUND: Poor or no clinical signs of psychological distress are usually observed in patients affected by severe or profound mental retardation (MR). The aim of this study was to use clinical and hormonal parameters in order to compare the amount of stress in patients affected by different degrees of MR undergoing general anaesthesia for dental care. METHODS: Nine patients affected by mild or moderate MR (group A: intellective quotient > 35) and 12 affected by severe or profound MR (group B: intellective quotient < 36), undergoing general anaesthesia for dental care, were studied. The reaction to venipuncture before anaesthesia induction and the recovery from anaesthesia were assessed by clinical scores. Cardiac rate and arterial pressure were recorded before and after venipuncture and after removing the endotracheal tube. Cortisol and prolactin, two hormones affected by stress, were determined in plasma the day prior to surgery (1), after anaesthesia induction (2), and after removing the endotracheal tube (3). RESULTS: During venipuncture, group B was significantly less reactive than group A (p < 0.05) and showed no arterial pressure increase while group A presented a significant increase of systolic pressure; conversely, the groups shared the same hormonal pattern at time 2 (cortisol did not change, prolactin increased significantly). Postoperatively A and B groups did not differ regarding cardiac rate and arterial pressure, while a wider range of postoperative scores was observed in group A in which a further prolactin increase was registered at time 3. CONCLUSIONS: Clinical evaluation can underestimate the stress to which patients affected by severe or profound MR are subjected during anaesthesia induction. PMID- 10479837 TI - Shortening the discharging time after total hip replacement under combined spinal/epidural anesthesia by actively warming the patient during surgery. AB - BACKGROUND: To compare passive thermal insulation by reflective blankets with forced-air active warming on the efficacy of normothermia maintenance and time for discharging from the recovery room after combined spinal/epidural anesthesia for total hip arthroplasty. METHODS: DESIGN: Prospective, randomized study. SETTING: Inpatient anesthesia at three University Departments of orthopedic surgery. PATIENTS: 50 ASA physical status I-III patients, who were scheduled for elective total hip arthroplasty. INTERVENTIONS: Patients received combined spinal/epidural anesthesia (CSE) with intrathecal injection of 15 mg of 0.5% hyperbaric bupivacaine. All procedures started 8-10 a.m., and operating room temperature was maintained between 21-23 degrees C, with relative humidity ranging between 40-45%. As warming therapy patients received either passive thermal insulation of the trunk, the two upper limbs and the unoperated lower limb with reflective blankets (group passive, n = 25), or forced-air active warming of the two upper limbs (group active, n = 25). Core temperature was measured before CSE placement (baseline), and then every 30 min until recovery of normothermia. RESULTS: Demographic data, duration of surgery, intraoperative blood losses, and crystalloid infusion were similar in the two groups. Arterial blood pressure decreased in both groups compared with baseline values, while no differences in heart rate were observed during the study. Core temperatures in passive group patients decreased more markedly than in actively warmed patients, with a 1 degree C difference between the two groups at the end of surgery (p < 0.0005). At recovery room entry seven patients in group active (24%) and 16 patients in group passive (64%) showed a core temperature < 36 degrees C (p < 0.01). Achievement of both discharging criteria and normothermia required 32 +/- 18 min in active group and 74 +/- 52 min in passive group (p < 0.0005). CONCLUSIONS: Forced-air cutaneous warming allows the anesthesiologist to maintain normothermia during combined spinal/epidural anesthesia for total hip replacement even if the convective blanket is placed on a relatively small skin surface with reflex vasoconstriction. Maintaining core normothermia decreased the duration of postanesthesia recovery and may, therefore, reduce costs of care. PMID- 10479838 TI - Gastroprotective effect of propacetamol against cold/restraint stress ulcers in rats. AB - BACKGROUND: Comparative evaluation of propacetamol and morphine on the cold restraint stress ulcers in rats. METHODS: The present study compared the effects of propacetamol hydrochloride (250 and 500 mg.kg-1 i.p.) and morphine hydrochloride (10 mg.kg-1 i.p.) against gastric mucosal damage induced by cold/restraint stress (4 degrees C for 3 h) in rats. Morphometrical and histomorphological studies were carried out. Mean ulcer number and length were calculated. RESULTS: The results show that propacetamol in the lower dose tested decreases the ulcer number and length by 56.4% (p > 0.05) and by 68.94% (p < 0.01). After propacetamol 500 mg.kg-1 the ulcer number and length were found significantly decreased by 74.83% and 83.5%. Marked decrease was found in morphine-pretreated group (-77.03% and -85.09%). The morphometrical results have been confirmed histomorphologically. CONCLUSIONS: It might be concluded that morphine (10 mg.kg-1) and propacetamol (500 mg.kg-1) are equipotent in their ability to prevent the stress ulceration in rats. PMID- 10479840 TI - Sepsis and organ dysfunction/failure. An overview. AB - Sepsis is a condition at high risk for the patients to develop organ(s) or system dysfunction/failure and represent a very limiting process for survival. Researchers and clinicians proposed standardization of terminology for sepsis and related problems to improve communication and to evaluate the efficacy of preventive measures and therapeutic interventions. Interrelationship among systemic inflammatory response syndrome (SIRS), infection and sepsis are surrounded by non infectious satellite events such as trauma, burns, pancreatitis, haemorrhagic shock, immune-mediated organ injury and infectious cause such as fungemia, parasitemia, viremia. The prevalence of infections among intensive care patients has been reported to vary from 15 to 40%. Usually indicators of sepsis are persistent hyperlactatemia and supranormal level of DO2. These conditions may progress as a sort of dynamic process known as endotoxaemia condition which is mediated by derangement of biohumoral factors inducing immunological dissonance and ultimately concomitant or sequential organs dysfunction/failure. Multiple sources of sepsis is a phenomenon clearly associated with poor prognosis and all the sepsis trials managed in the last decades have failed on reducing mortality rate in enrolled patients. Development of scoring system routinely used at bedside represent an important method to establish cost-effectiveness in this exiting area of study and clinical management. Controversial results on sepsis need a sort of consensus at different level from researchers to clinician experiencing new strategies for prevention and more appropriately therapeutic approach for the management of this syndrome. PMID- 10479839 TI - [Percutaneous or surgical trachetomy. Prospective, randomized comparison of the incidence of early and late complications]. AB - BACKGROUND: To compare early and late complications after either conventional surgical or percutaneous dilatational tracheostomy. DESIGN: Prospective, randomized study. SETTING: General intensive care unit and neuro-surgical intensive care unit in a university hospital. PATIENTS: 50 consecutive patients, requiring tracheostomy for prolonged mechanical ventilation. INTERVENTIONS AND MEASUREMENTS: Patients were randomly allocated to receive either surgical (surgical group, n = 25) or percutaneous dilatational tracheostomy (percutaneous group, n = 25). Occurrence of perioperative complication were carefully evaluated during ICU stay. Late complications were evaluated with both physical and endoscopic examination at 1, 3 to 6 months after tracheostomy. RESULTS: All surgical and percutaneous tracheostomies were successfully completed and no deaths directly related to the tracheostomy procedures were reported. Completion of the procedure required 41 +/- 14 min in the surgical group and 14 +/- 6 min in the percutaneous one (p < 0.0001). The incidence of early perioperative complications was higher in the surgical group (36%) than in percutaneous one (12%), (p < 0.05). The endoscopic follow-up demonstrated one segmental malacia and one stenosis of the trachea in the percutaneous group only (p = n.s.). Skin repair was better after percutaneous tracheostomy than in the surgical group (p < 0.01). CONCLUSIONS: In experienced hands, percutaneous dilatational tracheostomy is as safe and effective as the conventional surgical tracheostomy. The percutaneous technique is less time-consuming and has a lower rate of early infectious complications with better cosmetic results than the surgical technique. PMID- 10479841 TI - [Herpes Zoster and post-herpetic neuralgia: everything to revise?]. AB - The treatment and prevention of the algic symptomatology of Post-Herpetic Neuralgia (PHN) are problems still to be solved. In our opinion the reason of this failure is to be found in an insufficient knowledge of the pathophysiological phenomena which cause the PHN. In fact, many fundamental aspects, such as its "temporal" and symptomatological definition and its pathogenesis are still unsolved. This revision tries to focus the problems and discrepancies hindering an improvement in PHN treatment and prevention. The literature reports three different symptomatological levels after herpes zoster acute phase: patients affected by pain and/or allodynia, patients with "abnormal sensations" (anesthesia, paresthesia, dysesthesia, prickling, itching, burning, etc.,) and patients in "complete recovery". Only two studies have analyzed the final incidence of these symptomatologies. Therefore, it is absolutely unclear whether the patients in "abnormal sensations", that is with a symptomatology which is definitely less weakening than pain, have to be considered in PHN (patients with pain) or in "complete recovery". On the contrary, from a clinical and physiopathological point of view this symptomatological difference may have a great importance: the symptomatological non-distinction could be the cause of the disagreement concerning PHN incidence data; this distinction could differentiate and compare in a reliable way the effects of the various therapies reported in the literature. The lack of a symptomatological classification does not facilitate a verification of the physiopathological hypothesis of PHN onset and maintenance. The differentiation among pain, "abnormal sensations" and "complete recovery" could correlate these three symptomatological conditions to the anatomopathological and viral data which according to literature characterize the evolution of Herpes Zoster acute phase in PHN. This would largely help to improve the therapeutic strategy. PMID- 10479842 TI - [Hypertrophy of the lingual tonsil and difficulty in airway control. A clinical case]. AB - A male patient suffering for exertional angina was scheduled for coronary bypass. Physical examination was unremarkable except for oropharynx classified as Mallampati II. After anesthetic induction with fentanyl 10 micrograms/kg, thiopental 5 mg/kg and muscle relaxation with succynilcoline 1 mg/kg, the patient was ventilated via a face mask. Laryngoscopy revealed a bulky mass arising from the rigth base of the tongue hiding the epiglottis and all the vocal apparatus (Cormack class 4); a failed intubation caused bleeding. Facial mask ventilation became more difficult therefore, considering the task on managing the airway, a n. 4 laryngeal mask was positioned by the senior anesthetist. Two intubation attempts failed while ventilation via laryngeal mask became more and more difficult. Surgery was therefore cancelled due to inability to airway management. The mass, biopsied by an otolaryngologist, resulted to be a lingual tonsillar hyperthrophy and therefore was not removed. The patients was re-scheduled for cardiac surgery. Maintaining spontaneous breathing during light sedation, with topical anesthesia, this patient was successfully intubated over an Olympus BF P 10 bronchoscope. The patient had an uneventful operation, was regularly extubated and was discharged on the sixth postoperative day free from airway complications. Although we followed only some of the guidelines for the management of the difficult airway: a senior anesthetist was immediately called when an anatomic alteration was evident; progressive difficulty in maintaining the airway prompted the positioning of a LMA, the restoration of the spontaneous breathing and the cancellation of the elective operation had been mandatory when a class 4 Cormack was found at laryngoscopy. This situation requires an alternative approach to intubation or with the retrograde technique or with the aid of a fiberscope both maintaining spontaneous breathing. PMID- 10479843 TI - Electric nerve stimulation: analysis of two clinical cases of failed nerve electrolocation by using sheathed needles. AB - The aim of this work was to explain the reasons of two unsuccessful blocks of sciatic nerve even if anaesthetic solution was injected through insulated needle on elicited twitch. The clinical cases were two outpatients undergoing diagnostic arthroscopy of knee under anaesthetic block of sciatic and femoral nerves. In both patients, the muscular twitch appeared when the ischiatic bone was kept in unexpected touch with needle tip. In spite of the attempt to locate correctly the needle (the touch with bone means that the nerve is not in front of the needle tip), the injection of anaesthetic solution was unsuccessful. In clinical environment, when electroinsulated needles gathered total amount of administered current on the needle tip, it was not possible to elicit a twitch just at the moment of touch of the needle with the bone. Referred events disagree with some experimental works performed out of clinical environment, which found that total amount of administered current through an insulated needle gathers always in front of the tip. Our clinical observations seems to confirm an electrolocation mistake called "electrical shadow". The ability of sheathed needles to work as occasional capacitor due to the alternation of two conductor layers (needle shaft and tissue) and of a dielectric (coating material) can explain some missing electrolocations, as the appearance of electric fields within dielectric needle sheathing. PMID- 10479844 TI - [Use of the laryngeal mask for airway control in difficult intubations in children]. AB - LMA was introduced in clinical practice by Arthur Brain in 1983 as a valuable substitute of tracheal tube in adult who underwent general anaesthesia; since then its applications have been extensively studied. LMA is a relatively new non invasive ventilatory device which has allowed a radical change in the management of modern general anaesthesia. In this study, the application of LMA is assessed during induction and maintenance of general anaesthesia in children affected by severe facial deformities that could render the placement of the tracheal tube difficult. Three patients were affected by complex malformative syndromes involving the maxillo-facial skeleton and one patient presented a massive teratoma, originating from the orbit. In all these cases, LMA provided a patient airway and a satisfactory ventilation during both induction and the repeated attempts of inserting the tracheal tube; in one case, since the orotracheal intubation failed, LMA has proved to be as effective as the tracheal tube during the maintenance of general anaesthesia. Therefore, LMA is recommended as an essential ventilatory device in the hands of paediatric anaesthesiologists. PMID- 10479845 TI - [Acute salicylate intoxication after trancutaneous absorption]. AB - Topical salicylate preparations are primarily employed as keratolytic agents in the treatment of dermatologic disorders. A case of severe salicylate intoxication in a 70-year-old man with psoriasis, treated with a topical cream containing salicylic acid, is described. After five days the patient was admitted to ICU with encephalopathy and severe acid-base disturbances (respiratory alkalosis, metabolic acidosis, increased anion gap). A plasma salicylate concentration was elevated at 695 mg/Lt. Emergent hemodialysis was accompanied by a rapid lowering of plasma salicylate concentration and resolution of metabolic acidosis. Salicylate is well absorbed by normal and diseased skin. In this patient the lack of a normal epidermal barrier greatly enhances absorption of topical salicylate. It is therefore suggested that all topical salicylate treatments should be routinely monitored with salicylate blood concentration especially during the initial few days after onset or after any changes in treatment. PMID- 10479847 TI - [Acute pulmonary edema from contrast media. A clinical case]. AB - A rare and strong adverse reaction to contrast media (mdc) i.v. injected to carry out an abdominal vessels' angiography is reported. The patient suffered from non cardiogenic pulmonary edema and hypoxia. Chest X-ray detected the same radiological patterns of ARDS: in contrast alveolar edema became insignificant in 5 days. Serum immunocomplexes levels were high in the early phase of the reaction. Serum IgE, C3 and C4 were within the normal ranges. The case can be considered strongly significative because of the rarity of this syndrome; moreover, the early monitoring detected some data which could contribute to the knowledge of the physiopathology of contrast media adverse reaction. Overall, the presence of an immunological disease can increase the risk for adverse reactions to contrast media. PMID- 10479846 TI - [Pulmonary re-expansion edema. Description of two cases and observations about its management]. AB - Two cases of Reexpansion pulmonary edema (RPE), an uncommon complication of the treatment of chronic lung collapse secondary to pneumothorax or pleural effusion, are described. RPE is generally unilateral and occurs when the lung is rapidly reexpanded by active evacuation of large amounts of air or fluid. Nevertheless, both cases observed confirm that RPE can be seen when the pulmonary collapse is of short duration and the lung is reexpanded without suction. The pathogenesis of RPE is still unclear and is probably multifactorial. Implicated in the etiological process of RPE are chronicity of collapse, technique of reexpansion, increased pulmonary vascular permeability, airway obstruction, loss of surfactant, and pulmonary artery pressure changes. In the observed cases RPE appeared unexpectedly and dramatically, which is typical of the condition. The edema progressed for 24-48 hours, although it may persist for 4-5 days. Therapy was supportive and proportional to the severity of the clinical picture. Both needed mechanical ventilation, while only in case 1 was a hemodynamic support applied. Since the outcome is still fatal in 20% of cases, physicians treating chronic lung collapse must be aware of the possible causes and try to prevent the occurrence of this complication. PMID- 10479848 TI - [NMR in the diagnosis and treatment of neurologic lesions in decompression sickness]. AB - The case of a decompression sickness in woman, diving to 26 meter depth is reported. The patient was helped by instructor's computer (error!) and she presented risk factors for embolic disease (obesity, smoke, estroprogestinic therapy). She presented with many symptoms of decompression sickness during immersion and during re-ascent (headache, vertigoes and paresthesias). She was not treated on the place of incident, but only 36 hours later at our center of hyperbaric medicine. Her Magnetic Resonance imaging showed hyperintensity lesions of white matter. PMID- 10479849 TI - [Translaryngeal tracheostomy as modified by Sarpellon et al. A convicing alternative to the original method]. PMID- 10479850 TI - [The new law on organ transplantation]. PMID- 10479851 TI - Efficacy evaluation of prostaglandin E1 against placebo in patients with progressive systemic sclerosis and significant Raynaud's phenomenon. AB - BACKGROUND: Raynaud's phenomenon, due to connective tissue diseases, is difficult to treat successfully. Symptomatic improvement has been reported using nifedipine or iloprost, but adverse side effects may limit their use. The purpose of this study was to examine the effects of PGE1 (Alprostadil) in patients with scleroderma and severe Raynaud's disease. METHODS: Twelve females, aged 50-67 years, were included in the study with six of them receiving a 3-hour infusion of alprostadil at the standard dosage of 60 micrograms in 250 cc of physiological infusion for six consecutive days and the remaining six receiving placebo (250 cc of physiological infusion administered in the same manner). RESULTS: After infusion, blood flow, digitally measured by telethermography was increased only in patients treated with alprostadil. The number, frequency and severity of attacks recorded were reduced only in patients treated with alprostadil. No side effects were recorded during and after the infusion. CONCLUSION: In conclusion, alprostadil is effective in the management of Raynaud's phenomenon, due to scleroderma. PMID- 10479852 TI - Popliteal aneurysms: current management and outcome. AB - BACKGROUND: Peripheral artery aneurysm is the most common peripheral arterial aneurysm. Clinical pictures, perioperative diagnostic evaluations, surgical treatment and results are described. METHODS: From January 1982 to September 1998, 65 popliteal aneurysms in 48 patients were treated in the Department of Vascular Surgery of Busto Arsizio Hospital. The series comprises 44 men (91.7%) and 4 women (8.3%) with mean age 65 years (range 44 to 87 years). At the time of surgery, 38 of the 65 aneurysms (58%) were treated because symptomatic: 31 (82%) for limb ischemia (17 for acute ischemia, 14 for chronic ischemia), 6 (16%) for compression on the adjacent popliteal vein and 1 (2%) for rupture. Forty-seven (72.3%) aneurysms were treated electively: 27 cases were asymptomatic and 20 others had chronic leg ischemia or compression on the popliteal vein. The operations performed are listed as follows: 1 primary limb amputation for irreversible ischemia, 1 lumbar sympathicectomy and 62 leg limb revascularizations with graft interposition (vein or synthetic prosthesis). The patient with aneurysmal rupture underwent leg amputation. RESULTS: Mean follow-up is 6.5 years (ranging from 2 months to 10 years) and revealed important differences between symptomatic versus asymptomatic patients in limb salvage and graft patency: inferential analysis of amputation incidence is significant for p < 0.01 (Fisher exact test). CONCLUSIONS: We think that surgery is the best treatment also for small popliteal aneurysms, because of their frequent complications. PMID- 10479853 TI - [Surgical management of asymptomatic carotid artery stenosis. Authors' experience, problems and prospectives]. AB - BACKGROUND: Some trials have demonstrated effectiveness of carotid endarterectomy (CEA) for preventing stroke in patients with severe symptomatic carotid stenosis. Although some researches, indication to surgery for asymptomatic carotid stenosis is debated up today. Based on personal experience and literature, the main problems of CEA for asymptomatic stenosis are discussed. METHODS: DESIGN: Retrospective study. SETTING: Section of Vascular Surgery, University Department. PATIENTS: CEA was performed in a consecutive series of 63 cases with asymptomatic stenosis (59 patients, 40 males and 19 females, ages ranging from 46 to 80 years, mean 67.9). INTERVENTIONS: CEA was performed under general anesthesia, with primary closure of arteriotomy in 37 cases and patch angioplasty using PTFE in 24, using eversion technique in 2 cases. Pruitt-Inahara shunt was used in 10/63 cases (15.9%), according to the mean velocity of the middle cerebral artery at carotid clamping/mean velocity of the middle cerebral artery pre-clamping ratio x 100 equal to or lesser than 15%, evaluated with transcranial Doppler, or stump pressure lesser than 50 mmHg, when transcranial Doppler examination was not possible. MEASURES: Operative mortality and postoperative morbidity. RESULTS: Operative mortality plus postoperative stroke were 1.6% (1/63). Operative mortality was precisely 0.0%. Postoperative complications were two: one was a neurologic deficit (monoparesis of the arm) and the other was myocardial ischemia. CONCLUSIONS: Four main problems have been shown in CEA for asymptomatic stenosis: 1. Identification of asymptomatic stenosis: 2. Assessment of risk for stroke: 3. Role of CEA: 4. Questions about surgical treatment. For the first problem, it is important to consider possible indicators for carotid stenosis (contralateral carotid stenosis, coronary artery disease, aortic aneurysm, peripheral arterial disease, etc.). With regard to the second problem, it is important to know the natural history of the carotid stenosis, which shows a stroke rate of 1-2% per year. Regarding the third problem, the role of CEA is conditioned by: trials, patient conditions, lesion characteristics and ability of the surgeon. Further studies should identify some groups of patients (with severe carotid stenosis, dyshomogeneous plaque, progression of plaque, etc.), who can profit from CEA. Finally (fourth problem), CEA for asymptomatic carotid stenosis carries all common problems of carotid surgery (preoperative assessment, evaluation of cerebral ischemia due to carotid clamping, shunt, closure of arteriotomy, etc.). Some of these problems can receive ultimate solutions from some studies in next years. PMID- 10479854 TI - [Correlation between clinical-instrumental pattern in venous pathology (capillaroscopic classification of the medial peri-malleolar area)]. AB - BACKGROUND: The high incidence of chronic venous insufficiency makes it quite an ordinary topic in the outpatients departments of vascular units. Furthermore, due to the discrepancy between symptoms and Doppler c.w reports, it is necessary to face this problem with the help of very sensitive instrumental methods. Therefore, the main aim of this article is to find out which method, among those used in the outpatients department of phlebology, is sensitive enough to establish a relationship between clinical and instrumental patterns in venous diseases. METHODS: Fifty-one subjects were studied: 44 affected by venous disease and 7 healthy subjects. All patients underwent clinical examination followed by classification by Widmer criteria, Doppler c.w., infrared photoplethysmography and capillaroscopy of the medial malleolus. RESULTS AND CONCLUSIONS: Capillaroscopy demonstrated to have the highest sensitivity, allowing to find out pathological alterations in subjects complaining of symptoms and with normal Doppler c.w. findings. Furthermore, the findings of venular networks (pathological) in the malleoli of healthy subjects, made it necessary to modify the current capillaroscopic classification (based upon three types of pictures: normal, reticular and halo formations), into a new extended version based on five steps, with three types of reticular sub-classes, in which the different gauge of the venules is satisfactorily related to the symptoms referred by the patients. PMID- 10479855 TI - Primary cardiac lymphoma. A case report and review. AB - Primary cardiac lymphoma is classically defined as an extranodal non-Hodgkin's lymphoma exclusively located in the heart and/or pericardium. However, over the last few years, this definition has been extended to include other localizations on condition that these are clearly less important then a cardiac site, that must remain the first, during the illness course, and the most important for its entity. PCL is extremely rare in immunocompetent patients, accounting for 1.3% of all cardiac tumours and 0.5% of all extranodal lymphomas, but it has been encountered with increasing frequency in patients with AIDS or other severe immunodepressive syndromes. PCL is difficult to diagnose, especially during the early stage of the disease, because of its non-specific clinical manifestations, the limited possibility of using non-invasive diagnostic techniques, and difficulties or delays in applying invasive methods. The malignancy of its histotypes and its delicate location are responsible for its rapid and frequently unfavourable evolution. Successful treatment, which is mainly based on anthracycline-containing polychemotherapies, is heavily dependent on an early diagnosis. After a general review of the literature, the authors describe the clinical case of a patient with a PCL that had a secondary central nervous system location, treated with polychemotherapy and autologous peripheral blood stem cell transplantation. Emphasis is placed on the fact that it is more difficult to eradicate the disease from the central nervous system than from the heart. PMID- 10479856 TI - [Whole pancreatic transplantation and islet transplantation. Experiment notes]. AB - BACKGROUND: The results of pancreas transplantation have greatly improved in recent years. The path to further improvements goes through extensive experimental researches. METHODS: This study describes the effects of different procedures as hemodynamic asset and postoperative outcome. Twenty-nine swine underwent a total pancreatectomy, and were stratified into five groups. Group one (n = 5) served as control. Group two (n = 7) was autotransplanted. Group three (n = 6) and group four (n = 6) underwent allotransplantation; the first without immunosuppression and the second treated with cyclosporine and steroids. In group five (n = 5) Langerhans Islets transplantation was performed. RESULTS: Survival was different depending on which methodology was applied. The postoperative survival was 7 +/- 2 days in group one, 24 +/- 16 days in group two, 17 +/- 7 days in group three, 27 +/- 8 days in group four and 12 +/- 6 days in group five. CONCLUSIONS: The postoperative glucose control was normal in group two and group four while a severe diabetes appeared in group one (group 1 vs group 2: p < 0.05) and in group three during acute graft rejection after the 12th postoperative day (group 3 vs group 4: p < 0.05). Glycemia was slightly controlled in group five. The intraoperative hemodynamic status was evaluated at the time of pancreatectomy, harvesting, revascularization, and when surgery was over. Among the different parameters studied (mean arterial and pulmonary pressure, pulmonary wedge pressure, central venous pressure, cardiac output, oxygen extraction ratio, systemic vascular resistance, oxygen delivery and oxygen consumption), a statistically significant difference between group one and group five (p < 0.05) was observed. PMID- 10479857 TI - [Mechanical reconstruction after total gastrectomy. Analysis of results]. AB - BACKGROUND: Stapling devices introduction in gastroesophageal surgery has allowed better results. METHODS: An analysis is made of a consecutive series of 108 patients operated on with total gastrectomy and mechanical esophagojejunostomy for gastric neoplasm, in the last decade (1984-1995). RESULTS: The incidence of anastomotic clinical leakage was 2.4% with a perioperative mortality of 0.9% (1 patient) while the incidence of stenosis, treated in every patient with endoscopic dilatation, was 3.2%. These data are in accordance with previous reports. After an accurate analysis of the factors influencing the complication rate pointed out that in most of the cases a technical error in the anastomosis confection is present. CONCLUSIONS: The conclusion is drawn that the use of stapling devices in gastroesophageal surgery represents an elective indication. Nevertheless it is mandatory to respect the principles of visceral mechanical anastomosis after an adequate training. PMID- 10479858 TI - Surgical management of pancreatic pseudocyst. AB - BACKGROUND: Pancreatic pseudocysts were once considered to be an unusual complication of acute chronic and traumatic pancreatitis. METHODS: This work was made in order to study the results of the operative methods in 24 patients with acute chronic and traumatic pancreatic pseudocysts, treated by external or internal drainage during the years 1990-1995 at the Athens Red Cross Hospital and compare these results with those of international literature. Pain was the common symptom for all patients. Gallstones were the most important aetiological agent in thirteen of the 24 patients, while alcoholic pancreatitis was diagnosed in only 6 of them. Fifteen patients (62.5%) were treated by surgical drainage or resection and 9 patients (37.5%) were treated by observation, one by percutaneous and one by endoscopic drainage. The rest had small cysts (less than 5 cm) and were treated by observation. RESULTS: The most frequent complication of internal cyst drainage was upper gastrointestinal haemorrhage. The rate of mortality was 7%. CONCLUSIONS: Anatomical considerations dictate the choice of operation. Cystogastrostomy, for example is inappropriate unless the stomach is closely applied to the front of the cyst. We preferred cystojejunostomy Rouen-y because the Roux loop can be anastomosed to the lower part of the cyst. Cystoduodenostomy should be reserved for pseudocyst in the head of the pancreas. Resection is an alternative to internal drainage for chronic pseudocyst of moderate proportions, for those that have largely replaced a portion of the pancreas. PMID- 10479859 TI - 99mTC sulphur colloid phagocytosis by the mononuclear phagocytic system (MPS) of rats with portal hypertension. AB - BACKGROUND: Hypertension may be a cause of sepsis, especially among cirrhotic patients. METHODS: In the present investigation we studied the response of the mononuclear phagocytic system in 25 female Holtzman rats divided into five groups of 5 animals each: A, sham-operated animals; and animals with portal hypertension induced by calibrated ligature of the portal vein at different times of evolution: B, 15 days; C, 30 days; D, 45 days; E, 60 days. After the period of observation, 1 MBq/kg 99mTc colloidal sulfur was injected through the caudal vein and the animals were sacrificed 30 minutes later. Blood samples were obtained for GOT, GPT and albumin determination and radioactive counts, and liver, lung, spleen and bone marrow samples were obtained for the determination of the proportional phagocytosis index per gram organ. The results were analyzed statistically by the Kruskal-Wallis test. RESULTS: Increased radioactivity was observed in bone marrow and spleen and higher GPT levels were detected after 15 days. A reduction of splenic phagocytosis, an increase of pulmonary phagocytosis and increased GPT levels occurred after 30 days. In the 45-day group there was an increase in non-phagocytized particles remaining in blood, and in the 60-day group there was only and increase in GPT values. CONCLUSIONS: We conclude that portal hypertension causes phagocytic alterations during the early phases in this experimental model. PMID- 10479860 TI - [Behavior of Helicobacter pylori in 3d and 4th age-bracket patients]. AB - AIMS AND BACKGROUND: Helicobacter pylori is known to be the pathogenic agent of atrophic gastritis and above all ulcer disease. It is also thought to play a role, together with other factors, in the development of stomach cancer and is currently the focus of numerous studies. In industrialised countries, infection is relatively commonplace in children, whereas its incidence rises more steeply in patients aged over 35, reaching a mean prevalence of 50% in over 60 year olds. METHODS: During 1997 the authors carried out an epidemiological study to assay levels of anti-Helicobacter pylori (IgG anti-Hp) in a sample of 91 aged patients hospitalised in the S. Margherita Geriatric Hospital in Pavia. The patients had been admitted to hospital for various pathologies and the study also took into account gastric pathologies which usually, particularly in neoplastic forms, prefers the third and fourth ages. The assay for antibodies was performed using mouse monoclonal antibody, specific for Helicobacter pylori, absorbed on a polyester pad. A total of 5 ml of blood were collected from each patient. The blood samples were all centrifuged and the serum frozen at -20 degrees C until the tests were performed. RESULTS: In this study, 46 of the 91 patients tested were positive for anti-Helicobacter pylori antibodies, 35 were negative and 10 borderline. CONCLUSIONS: The high level of positive results found in these patients is in line with the findings reported by other authors. PMID- 10479861 TI - [Giant benign gastric ulcer penetrating into the liver, pancreas and mesocolon]. AB - Penetration of the liver, pancreas and transverse mesocolon by a giant benign gastric ulcer is relatively uncommon, and literature contains a few reports of this complication. The preoperative histological diagnosis may be difficult or impossible. A 63-year-old female patient with a history of seven months of lack of appetite, asthenia, epigastric pain, a remarkable weight decrease, presenting at physical examination a large, smooth margins, not pulsating, quite fixed abdominal mass, is reported. Echography confirmed the presence of a mass of approximately 14 x 19 cm, with solid and liquid content. Biopsy showed inflammatory elements and cellular detriti. Barium enema showed that the mass compressed the descendent colon, which appeared dislocated. Tumor markers (CEA, CA 19-9, alpha-fetoprotein) where in the normal range. Endoscopy showed a giant angular ulcer whose bottom was represented by necrotic material (after the definitive histological examination it proved to be hepatic tissue). At TC scan of the abdomen, a remarkable thickening of the gastric wall was present. At surgery the stomach appeared increased in volume, with remarkably thickened walls, tenaciously sticking to II and III hepatic segments, to the pancreas and transverse mesocolon. A total gastrectomy was performed because of the depth of the ulcer penetration and the extension of the alteration of the gastric wall, even if the giant gastric ulcer, in the literature, is more frequently benign than malignant. PMID- 10479862 TI - Bilateral lumbar hernia associated with abdominal hernias. A case report. AB - Multiple abdominal hernias constitute a quite rare event in surgical practice. Lumbar hernias are even rarer and, to our knowledge, this is the first report in the English literature of its association with abdominal hernias. A case of multiple abdominal hernias, namely an epigastric hernia and a left inguinal hernia together with a bilateral lumbar hernia in a 65-year-old man attending the Out-patient department of Hoima Hospital-Uganda is described. A one-stage repair under local anaesthesia was chosen. The patient recovered uneventfully and no recurrence was noted at 1 year follow-up. This case report supports that a "one stage" procedure under local anaesthesia can be appropriate as surgical treatment of multiple abdominal hernias when abdominal defects are of a small size. A minimal surgical approach to lumbar hernias seems also suitable when transverse muscle aponeurosis defect can be carefully repaired and covered with adjacent muscles. PMID- 10479863 TI - [Spigelian hernia: anatomo-clinical considerations]. AB - Two cases of Spigelian hernia are reported. Starting from the analysis of these cases, some observations about the anatomy, the etiology, clinic, diagnostic methods and the best therapeutic choices now available for this uncommon pathology are presented. PMID- 10479864 TI - [A case of intestinal obstruction after migration of esophageal endoprosthesis]. AB - Endoscopic dilatation and stenting are the procedures of choice in the treatment of oesophago-gastric anastomotic stricture following oesophagectomy, leading to immediate relief of dysphagia with a low procedure-related risk. Prosthesis displacement is a described late complication, easy detected and sorted out whenever possible with endoscopic retrieval. A case in which stent migration ended at the terminal part of the small intestine, presenting with the clinical features of mechanical ileum, is described. PMID- 10479865 TI - [Massive thrombosis of the superior mesenteric artery following splenectomy. A coincidence?]. AB - Postsplenectomy thrombocytosis is a well recognised complication about which there is little published information. Therefore, postoperative complications of this effect have not been emphasised. An experience with a case of superior mesenteric artery thrombosis and small intestinal ischemia following splenectomy is reported. We reviewed the literature, but we wasn't able to find any case, whereas the association between splenectomy and mesenteric or portal vein thrombosis is well known. Pathogenesis of postsplenectomy thrombocytosis is poorly understood. Theories to explain it include removal of the splenic sequestration effect or removal of a regulatory humoral factor produced by the spleen. Both mechanisms could be operative at the same time, explaining the observation that some patients develop thrombocytosis related complications soon after surgery, while others after a longer time. The literature on the thromboembolic risk of postsplenectomy thrombocytosis is inconclusive and no studies have established whether patients with thrombocytosis following splenectomy should be treated with anticoagulants or antiplatelet medications in order to prevent thrombotic complications. Certainly, a recommendation for the routine use of these drugs cannot be made on the basis of one observation, but the need for controlled studies must be stressed. PMID- 10479866 TI - [Bochdalek's diaphragmatic hernia in adults. Description of a clinical case]. AB - Bochdalek's hernia is a congenital posterior diaphragmatic defect, resulting from persistence of the pleuroperitoneal canal of embryonic life. Bochdalek's hernias are rare occurrences in adult patients and they present commonly with digestive symptoms and need surgical therapy. This article describes a case of an 88-year old female with symptomatic right Bochdalek defect containing intra-abdominal viscera with no hernial sac. PMID- 10479867 TI - [Perforation of primary non-Hodgkin's lymphoma during chemotherapy. Clinical evaluation and therapeutic strategy]. AB - Primary cecal lymphoma are very rare. The case is reported of an elderly woman having voluntary preliminary chemotherapy with consequent cecum perforation due to tumoral necrosis, some days after the first chemotherapy. The diagnostic, clinical and therapeutic protocol is analysed and stress is laid on the rare clinical cases described. Resection is advisable in the still delimited forms in order to avoid surgical complications due to preliminary chemotherapy and in order to classify precisely the neoplasm. PMID- 10479868 TI - [Primary hyperparathyroidism and acute pancreatitis. A rare clinical association]. AB - Hyperparathyroidism and hypercalcaemia are considered to be a rare cause of acute pancreatitis. The relationship between hyperparathyroidism and pancreatic inflammatory disease remains controversial, but it may be related to the translation from inactive to active trypsinogen by hypercalcaemia. Surgical correction of parathyroid disease and normalization of serum calcium levels may ameliorate the acute pancreatitis. Also the mechanism of pathologic zymogen activation during acute pancreatitis remain unknown; probably the pancreatic "autodigestion" is the result of anomalous intracellular transport of secretory proteins activated by lysosomal hydrolases. A case of acute pancreatitis and hyperparathyroidism due to solitary parathyroid adenoma occurred in a 66-years old woman is reported. After the excision of parathyroid adenoma the serum calcium levels and the function of the pancreas returned to normal. This suggests a cause and effect relationship between hyperparathyroidism and acute pancreatitis. PMID- 10479869 TI - [Acquired deficiency of von Willebrand factor (wWF) associated with intestinal angiodysplasia. An unusual clinical case]. AB - A rare case of acquired von Willebrand's disease associated with intestinal angiodysplasia is described. The case is very interesting because it clinically mimicked an intestinal neoplasm. PMID- 10479870 TI - [Muscle metastasis of bronchopulmonary carcinoma]. AB - Typical sites of bronchogenic carcinoma metastases are liver, brain, bones or adrenal glands. Rarely and in advanced dissemination phase it could involve the skeletal muscle. Two cases of metastases in the skeletal muscle from bronchogenic carcinoma, one of which revealed this neoplasia, are reported. PMID- 10479871 TI - [Pregnancy cholestasis. Clinical aspects and neonatal outcome]. AB - AIM: Gravidic cholestasis is a syndrome that is usually manifested during the third trimester of pregnancy and regresses in puerperium. It is characterized by the onset of itch, with or without associated jaundice and alterations of hepatic functional parameters. Its incidence varies according to geographical area. METHODS: A retrospective analysis was made regarding the frequency of this pathology in pregnant patients attending the Gynecology and Obstetrics Clinic of Trieste from 1-1-1980 to 31-7-1997. The epidemiological and clinical characteristics were studied relating to the course of pregnancy and neonatal outcome. Patients were identified on the basis of diagnosis on admittance and anamnestic and laboratory data, generalised itch and increased transaminase, biliary salts and alkaline phosphatase. Patients suffering form active-phase viral hepatitis were excluded, as were those in whom symptoms appeared after the start of alpha methyldopa administration. RESULTS: The frequency of gravidic cholestasis in this series was 0.36%. No significant differences were recorded in terms of age, parity and weight increase in these patients compared to the general population of pregnant women. Birth was spontaneous in 66% and by cesarean section in 34%. The frequency of premature births was 12%. The Apgar score at 5 min was satisfactory in all neonates. CONCLUSIONS: Careful monitoring of pregnancy associated with suitable obstetric care enabled a zero rate neonatal and maternal mortality to be achieved. PMID- 10479872 TI - [Cystic adenomatoid malformation of the lung at the fetal stage. Ultrasonographic diagnosis and counseling]. AB - BACKGROUND: The aim of this study was to report the series of cystic malformation of the lung (CAML) detected at the Center for Prenatal Diagnosis of the University of Naples Federico II and compare the data with the current literature. METHODS: In the period 1994-1997, ten cases of CAML have been detected at our unit. For each case, the following parameters were recorded: size of the cysts (according to the Stocker classification), side, presence of mediastinal shift, hydrops. For confirmation, necropsies and post natal files were sought. RESULTS: CAML was type I in 3 cases, type II in 3, and type III in 4 cases. mediastinal shift was present in 7 cases, hydrops in 2 and bilateral renal agenesis in 1. Seven cases underwent termination of pregnancy, 2 are currently alive and thriving after surgery and one pregnancy is ongoing at 36 weeks of gestation. The diagnosis was straightforward in all cases, and there were no problems of differential diagnosis. CONCLUSIONS: From an analysis of the data published in recent literature, it can be affirmed that hydrops but not mediastinal shift have negative prognostic significance. The abortion rate of 70% in our series, including 4 cases with a good prognosis, is somewhat frustrating. This figure underlines the need for a multidisciplinary counseling in which the pediatric surgeon, the psychologist and the sonographer may support the couple in overcoming the stress related to the acknowledgement of the lesion in order to reach the final decision about the outcome of the pregnancy. PMID- 10479873 TI - [Clinical considerations on oligohydramnios]. AB - BACKGROUND: Oligohydramnios represents a physiopathologic process, associated to a high rate of pregnancy complications and increased fetal morbidity and mortality. The purpose of this study is to determine whether amniotic fluid ultrasound evaluation is a useful parameter to identify pregnancies at risk for fetal distress. METHODS: 91 pregnant women with oligohydramnios were followed, and fetal conditions monitored with cardiotocography and ultrasounds. Delivery way and new-born conditions at birth were considered too. RESULTS: Cardiotocograms appeared abnormal in 61 cases (67%). Intrauterine growth restriction (IUGR) observed in 36 cases (39.56%). Cesarean section was performed in 60 cases (66%) for fetal distress, identified with cardiotocography. Also significant was the correlation with oligohydramnios, IUGR and cardiotocographic anomalies. From the results, it may be inferred that oligohydramnios is an index of affected fetal good health, because frequently is related with pregnancy anomalies and fetal health involvement. CONCLUSIONS: We can then affirm that the ultrasound evaluation of amniotic fluid represents a useful means to identify pregnancies at risk for fetal distress. In case of oligohydramnios, it is important to increase the surveillance on fetus, with the different means at our disposal (ultrasonography, velocimetry, cardiotocography). PMID- 10479874 TI - [Analysis of decline in seminal fluid in the Italian population during the past 15 years]. AB - BACKGROUND AND AIMS: The authors wished to study whether there has been a real decline in the quality of semen over the past years, as reported in the literature by a number of studies. This paper analyses the basic parameters of semen from 1068 males who have acted as donors during the past 15 years (1981 1995) for our semen bank. METHODS: Tables have been drawn up showing the number of cases examined every year, the mean concentration of spermatozoa, the mean percentage of mobile sperm and the mean percentage of non-conforming elements. RESULTS: An analysis of the data reported here shows that the mean concentration of spermatozoa in semen has shrunk from 88 x 10(6)/ml in 1981 to 61 x 10(6)/ml in 1995, a reduction of 30.7%; mean total motility has diminished from 74 to 66%, whereas the mean percentage with typical morphology has fallen from 76 to 63%. No statistical analysis was performed since variance analysis could not be performed on the sample in question to isolate the various sources of error. CONCLUSIONS: In conclusion, there has been a marked fall in the quality of semen over the past few years in individuals who regard themselves as fertile and are selected as semen donors. In the discussion the authors examine the various hypotheses regarding the possible causes of this deterioration in quality, opting for the argument based on the increased use of pollutants during the period in question in urban and rural areas. PMID- 10479875 TI - [Pharmacology of ritodrine]. AB - Tocolytic drugs are used in order to delay or to abolish preterm labour. These drugs inhibit uterine contractions and have different pharmacological properties. The most used tocolytic agents today are beta 2-adrenergic agonists (isoxuprine, ritodrine). The pharmacological actions, pharmacokinetics, toxicological and clinical aspects of ritodrine, a synthetic beta 2-adrenergic agonist with tocolytic properties are described. Findings of many clinical trials are also discussed. PMID- 10479876 TI - [Diagnosis and treatment of a case of adenocarcinoma of the cervix in pregnancy]. AB - The authors report a case of mucinous-papilliferous adenocarcinoma of the cervical canal in a pregnant woman. After performing three Pap tests which demonstrated the presence of normoconformate endometrial cells partly with a normal conformation and partly with nuclear atypia, the patient was monitored with periodical obstetric examinations. A neoformation of the portio was observed at week 28: differential diagnosis was made between a hypertrophic benign tumour and a well differentiated adenocarcinoma. The patient was monitored until birth and colposcopy was then performed with biopsy about one month later. Histological tests were positive for mucinous papilliferous adenocarcinoma. The patient underwent conisation with rotating loop and subsequently Piver 2 surgery owing to the suspected spread of the neoplasm beyond the margin of resection. The operative finding was negative and the patient is now free of disease. PMID- 10479877 TI - Primary abdominal pregnancy. A case report. AB - Abdominal pregnancies are very uncommon; in the United States they are seen once every 10,000 births and consist of approximately 1% ectopic gestations. We report one case with a primary pelvic-peritoneal ectopic pregnancy, diagnosed by chance at 11 gestational weeks complicated by hemoperitoneum and acute abdomen. This case is interesting because the early diagnosis of abdominal pregnancy is frequently difficult. In fact the clinical history, physical examination, laboratory and ultrasonographic findings are non specific. The physical examination is inconclusive in most patients. Findings such as abdominal tenderness, a closed uneffaced cervix, palpation of a pelvic mass distinct from the uterus are described as being suggestive of the abdominal pregnancy. Once the diagnosis of abdominal pregnancy is made, management of these patients requires a careful and further evaluation. Most clinicians agree that immediate operative intervention is indicated for those pregnancies prior to 23 to 24 weeks, because of the high incidence of maternal morbidity with significant risks of maternal mortality, in light of the poor prognosis for the fetus. In patients who present after 24 weeks, debate has arisen in the literature concerning the appropriateness of a more conservative approach. PMID- 10479878 TI - [A zinc sulfate and usnic acid preparation used as post-surgical adjuvant therapy in genital lesions by Human Papillomavirus]. AB - BACKGROUND: To evaluate the efficacy and safety of intravaginal administration of a zinc sulphate and usnic acid compound as adjuvant therapy of Human Papillomavirus (HPV) genital infection, after radiosurgical treatment (RS). METHODS: One hundred patients affected by HPV genital infection were enrolled in the study from October 1996 to July 1998. Patients were classified according to colposcopic and cytologic criteria and treated with RS. Patients were randomized into three groups: the first group did not follow any therapy after RS (control group), (n = 50); the second group was pharmacologically treated with intravaginal administration of a usnic acid and zinc sulphate compound (Zeta N, Bergamon Italia) before and after RS (n = 25), the third group was pharmacologically treated only after RS (n = 25). The last two groups were considered together for the statistical analysis. Patients were reevaluated after one, two, three and six months from electrocoagulation. The safety of treatment was also investigated. RESULTS: One month after RS. HPV lesions disappeared in 93% of the patients in the control group and in 100% of patients treated with usnic acid and zinc sulphate. After one month, reepithelization was complete in 65% of cases treated with usnic acid and zinc sulphate and in only 28% of the control group (p = 0.001). Two months later reepithelization was 94% in the patients pharmacologically treated compared to 76% of the control group (p = 0.06). Treatment prior to RS resulted in a reduction of the overall area of lesions in 88% of cases. Three months after RS, there was a significant reduction of recurrence in the group treated with usnic acid and zinc sulphate (p = 0.01). This reduction was still significant at six months (p = 0.005). CONCLUSIONS: Usnic acid and zinc sulphate adjuvant treatment improved time of reepithelization and reduce the recurrence with few side effects and a good compliance. PMID- 10479879 TI - [Usefulness of long-term follow-up of patients operated on for cryptorchidism]. AB - BACKGROUND: Cryptorchidism, testicular ectopia and rectractile testicle are different pathologies with conceivable different prognosis. As a matter of fact spermiogram used in these patients for the prognosis is really worse in bilateral cryptorchid than ectopic patients, because the tubular damage is present in the former than in the latter. It does not mean, anyway, that direct correlation exists between age of the patient at the operation and fertility. METHODS: 171 patients operated on for cryptorchidism or testicular ectopia at the Pediatric Surgery Department of the Regional Hospital of Treviso during the years 1974-1983 under-went follow-up at the age of 18 years with clinical evaluation, hormonal dosage (testosterone and gonadotropin after stimulus as GnRH) and spermiogram. RESULTS AND CONCLUSIONS: The present study does not differ from many other studies about hormonal levels and spermiograms of cryptorchid patients; it underlines long term follow-up utility of patients operated on for cryptorchidism and stresses the need of an eventual psychological evaluation of these patients. PMID- 10479880 TI - [Oxybutinin-desmopressin association in the treatment of primary nocturnal enuresis with diurnal urination disorders]. AB - BACKGROUND: Since most of the children observed in our Centre present enuresis with voiding disturbance, we carried out a study where these patients were treated with the DDAVP + Oxybutinin association. METHODS: We have treated 89 children with enuresis and voiding disturbances (urge incontinence, voiding urgency, urinated > 7 times a day), administering a drugs combination of desmopressin (20 micrograms/daily) and oxybutinin (0.3-0.6 mg/kg/bid or tid) for a variable period, depending on response to the treatment. RESULTS: The results demonstrate the efficacy of this association: we have observed a reduction in average bed wetting nights from 23.4 nights/month to 6.4 wet nights after 1 month, to 3.8 ad 2.9 respectively after 3 and 6 months from the beginning. Moreover we obtained a recovery of all daily voiding disturbances after 3 months. Fifty percent of children were cured after 4 months of therapy and finally 93.2% recovered at 6 months. On the other hand, 2 children were no-responders to the therapy even after 9 months of combined drugs administration. CONCLUSIONS: Based on these results we can affirm that children with nocturnal enuresis and voiding disturbance can be treated with this combined therapy. Actually, the reduction of urinary output and thus lower bladder filling, due to the desmopressin, decreases the onset of uninhibited bladder contractions and enhances oxybutinin activity. PMID- 10479881 TI - Partial cryptogenetic epilepsy and food allergy/intolerance. A causal or a chance relationship? Reflections on three clinical cases. AB - The possibility that certain foods or allergens may induce convulsions has already been reported in the literature. None of these studies has, however, shown a close correlation between food allergy and epilepsy, most reports being anecdotal and open to various aetiological hypotheses. The present report concerns 3 children with cryptogenetic partial epilepsy, diagnosed by means of electroencephalography, with behavioural disorders (hyperactivity, sleep disorders and writing difficulties). In these patients, instead of using anticonvulsive agents, treatment was based upon a cows milk-free diet, working on the hypothesis that there could be a casual relationship between intolerance to this food item and the epileptic symptoms. An improvement was observed in the children's behaviour and moreover, the electroencephalographic anomalies disappeared. Upon double blind oral provocation tests, these patients did not present an immediate reaction, but only after a few days. starting the controlled diet again led, in all cases, to disappearance of the electroencephalogram. In conclusion, it would appear feasible to hypothesize the role of food intolerance in the onset of convulsive crises, even if limited to certain types of epilepsy such as the cryptogenetic partial form. PMID- 10479882 TI - Lipoblastoma and lipoblastomatosis in children. AB - Lipoblastoma and lipoblastomatosis are rare mesenchymal benign tumors of embryonal white fat. They are found primarily in children less than 5 years of age. This tumor presents in two forms: a localized well-circumscribed lesion (lipoblastoma), or a multicentric type (lipoblastomatosis). The authors presents three cases, two of them with a buttock lesion, the last with a paravertebral one. It is recommended a complete but conservative excision of the tumor because there is a natural tendency to involution, although in the first year of life a local reexcision for recurrence is sometimes described. PMID- 10479883 TI - Cystatin C in paediatric nephrology. Present situation and prospects. AB - Cystatin C is a small basic protein with a MW of 13,359 Daltons, consisting of a non-glycosylated polypeptide chain containing 120 amino-acid residues. Cystatin C is produced in all the nucleated cells of the human body and its output rate is constant. The kidney is the main catabolic site of cystatin C, since the protein, by virtue of its low MW and its positive charge at normal pH, is freely filtered by the glomerulus and almost completely reabsorbed, catabolised and broken down in the cells of the proximal convoluted tubule. It is practically entirely filtered via the glomerular membrane, without any significant tubular secretion. The constant production rate of cystatin C in all the tissues, its elimination via the glomerular filter and its non-dependence on many extrinsic factors, including sex, age, diet, inflammation, are potentially ideal conditions for an endogenous biochemical marker of glomerular filtration. A recent method for determining cystatin C, is based on an immune reaction, could increase its clinical application. Not many studies have been conducted to date on cystatin C in children. The cystatin C concentration was higher during the first few days of life (range: 1.64-2.59 mg/L) with a rapid reduction during the first 4 months. Beyond the first year of life, cystatin C concentration became constant, with a reference range of 0.7-1.38 mg/L. On the basis of the data currently available, neonatal serum cystatin C would appear to derive from the newborn itself. In fact no correlations were found between maternal and neonatal serum cystatin C values. Cystatin C determination appears to be at least equivalent to serum creatinine measurement for the assessment of glomerular filtration rate in children. Further extended studies are needed to investigate these aspects more thoroughly in neonates. PMID- 10479884 TI - [Pediatrics and puericulture in the Renaissance. II]. PMID- 10479885 TI - [Genetic factors which regulate alcohol drinking behavior and their effects on health status]. AB - High alcohol sensitivity common among Orientals is mainly due to genetic polymorphism in the low K(m) aldehyde dehydrogenase (ALDH2) gene. The relation of the ALDH2 genotype to alcohol sensitivity and drinking behavior was investigated in a Japanese occupational population. The frequency of alcohol-associated symptoms generally increased in the order of the typical homozygote, heterozygote, and atypical homozygote. Both drinking frequency and amounts of alcohol consumption were also significantly affected by the polymorphism. Polymorphism in the alcohol dehydrogenase beta-subunit (ADH2 gene) appeared to contribute to skin flushing post-alcohol exposure but not to alcohol drinking behavior. Multivariate analysis revealed that high alcohol consumption, the ALDH2*1/*1 genotype, and high daily hassles levels significantly contribute to the prevalence of those with a high problem-drinking score in an occupational population. In the study to assess the effects of the ALDH2 polymorphism and alcohol use on the induction of chromosome alterations in peripheral lymphocytes, we found that lymphocytes from habitual drinkers with the atypical ALDH2 genotypes had significantly higher frequencies of sister-chromatid exchange (SCE) than those from the typical ALDH2 genotype. We also measured acetaldehyde reversibly bound to hemoglobin (HbAA). In volunteers with the ALDH2*1/*2 genotype, the HbAA levels increased immediately after the drink and the elevated levels persisted up to 48 h. Among male workers, HbAA levels were significantly correlated with the recent alcohol consumption levels in both the ALDH2*1/*1 and ALDH2*1/*2 genotypes. However, the slope was much steeper in the ALDH2*1/*2 than in the ALDH2*1/*1. SCE and HbAA may be utilized as a good biomarker for health problems in the atypical ALDH2 genotype. Further extensive studies are required for evaluation of the interactive effects of genetic and environmental factors on alcohol-related health problems. PMID- 10479886 TI - [Allylnitrile-induced behavioral abnormalities and findings relating to the mechanism underlying behavioral abnormalities]. AB - Nitriles are widely used in industry as plastics, solvents, and synthetic intermediates. It has been shown that the thermal degradation of acrylonitrile based plastics leads to the emission of a great variety of nitriles. Exposure of humans and experimental animals to some nitriles has been shown to lead to disorders of the central nervous, hepatic, cardiovascular, renal, and gastrointestinal systems. Iminodipropionitrile has long been known to induce in experimental animals behavioral syndromes that other nitriles have not been reported to induce. Recently, we have found that a single administration of allylnitrile, an analog of acrylonitrile, induces in rodents behavioral abnormalities including head twitching, head weaving, random circling, increased locomotor activity, backward pedaling, pivoting, and somersaulting. The induced abnormalities were persistent. Crotononitrile and 2-pentenenitrile also are able to produce behavioral abnormalities. Thus, the nitriles appear as a new class of neurotoxic compounds with potential relevance to the human health. The mechanism by which allylnitrile induces and maintains the behavioral abnormalities is summarised below. 1. Allylnitrile activates the serotonin (5-HT) system in the central nervous system, and as a consequence activation of 5-HT-2 receptors due to increased 5-HT may lead to induction of head twitching. 2. Although the data available indicate that the dopamine (DA) system may be involved in allylnitrile induced behavioral abnormalities, it remains unknown how the DA system relates to the abnormalities. 3. Allylnitrile decreases the noradrenaline level in the central nervous system, which is thought to be secondary to the 5-HT system activation mentioned above. The allylnitrile-induced head twitching, however, may occur in consequence to both enhanced beta-adrenoceptor stimulation and to the removal of tonic inhibitory control by alpha-2-adrenoceptors. 4. The neuropathological data indicate an important role of the medial habenular and raphe nuclei in allylnitrile-induced behavioral abnormalities. Onset of the behavioral abnormalities appears to be associated with the impairment in the medial habenulo-raphe relay owing to activation of apoptotic cascade in neurons. 5. On the basis of the findings with iminodipropionitrile and crotononitrile, allylnitrile might produce pathological changes in the vestibular sensory hair cells. Further studies are needed to explore the mechanism underlying the allylnitrile-induced syndromes. PMID- 10479887 TI - [Building aging level indices based on ability to perform personal care and household management activities]. AB - Defining aging as deterioration of the ability required for the activities of daily living with increasing age, we developed a formula for estimating the age of aging. In 1994 and 1995, a questionnaire survey was conducted for 11,592 individuals (4,885 men, mean age 61.6, standard deviation 14.6; and 6,677 women, mean age 63.2, standard deviation 14.8) who were members of 7 cohorts (5 community-based cohorts and 2 cohorts of examiners at a health-promotion center). The questionnaire included three groups of questions related to medical treatment, aging-related symptoms, and personal care (ADL; Activities of Daily Living) and household management (IADL: Instrumental ADL). Multiple regression analysis was made by sex and age group (over 65 years old, under 65 years old) using age as a dependent variable and the three question categories as explanatory variables. Using multiple regression analysis by question category, five items were abstracted from each of the three groups, so that a total of 15 items were abstracted from all questions. Five items were then abstracted from the 15 by multiple regression analysis, and the predicted aging age for an individual is estimated using this statistical model from the results of the questionnaire survey. The predicted aging age is significantly associated with age (r = 0.40-0.49, p = 0.0001). The difference between the predicted aging age and age is greater among the older or younger people. The expected predicted aging age is estimated using regression analysis of the predicted aging age on age. Aging level indices by sex and age groups were determined by the difference between the expected predicted aging age and age. We are planning to carry out an epidemiological study on the risk factor for aging using the aging level indices in seven cohorts. PMID- 10479888 TI - [Serum leptin level among non-obese students: relationship to body fat, blood pressure, serum lipids, physical activity, and eating habits]. AB - Leptin, a product of the ob gene, is thought to play a key role in the regulation of adiposity. However, it is unclear in humans as to whether or not leptin influences the blood pressure, serum lipids, physical activity level, or eating behavior in relation to obesity. Recent reports have indicated both gender-based differences in the leptin levels and a correlation of the percentage of body fat with leptin levels has been observed among obese subjects. As far as we know, these relationships among non-obese young adults have yet to be studied. Therefore, the serum leptin concentrations among 107 non-obese students (72 males and 35 females) were measured by a radioimmunoassay(RIA). Fasting leptin levels ranging between 1.2 and 23.4 ng/mL were observed in all subjects, and the levels among females were 2.6 times higher than in males (7.64 vs 2.95 ng/mL; p < 0.001). A close correlation was observed regarding the log-transformed leptin levels with the percentage of body fat as determined by the bioelectrical impedance analysis method(r = 0.734, p < 0.001 in males and r = 0.579, p < 0.001 in females). In conclusion, these data thus suggest the serum leptin levels among non-obese students show significant gender-based differences while, in addition, the leptin levels also correlate positively with the percentage of body fat. However, it remains unclear as to whether or not the leptin levels are independently related to the blood pressure, serum lipids, physical activity level (sports activity and leisure-time activity), and eating behavior (eating breakfast, mid-day snacking and nighttime snacking). PMID- 10479889 TI - [Preventive effect of selenium against the testicular injury by cadmium]. AB - An experimental study was performed on orchiopathy (testis disorder) by using cadmium (Cd) and on the prevention of orchiopathy by the administration of selenium (Se). By a single administration of Cd 1.4 mg/kg (12.4 mumol/kg) or a second administration of Cd 1.4 mg/kg 24 hr after the administration of Cd 1.0 mg/kg (8.9 mumol/kg), the testis of a mouse showed ex-tensive necrosis, and an extreme decrease of glutathione (GSH) concentration accompanied by an increase of thiobarbituric acid reactive substances (TBARS). When Se 1.4 mg/kg (17.7 mumol/kg) was given at the same time as Cd 1.0 mg/kg, the disorder was completely prevented and their serum Cd and Se concentrations were 1165 +/- 268 ng/ml and 534 +/- 128 ng/ml, respectively. However, when Se was given, separately from the Cd administration, either 24 hr or 72 hr before Cd administration, no effect to prevent testis disorder was found. On the other hand, when Se 1.4 mg/kg and Cd 1.0 mg/kg were given simultaneously and then Cd 1.4 mg/kg was administered 24 hr and 72 hr after the simultaneous injection, respectively, there was no sign of disorder caused by the second administration of Cd. When Cd was given after administration of Cd and Se, Cd concentration in the testis (0.88 +/- 0.078 microgram/g and 0.77 +/- 0.03 microgram/g) was about twice as much as the concentration in the case of no administration of Se (0.30 +/- 0.04 microgram/g). The testicular dysfunction could not be explained by the increased Cd concentration in the testis. The groups with high Cd concentration in the testis were accompanied by an increase in metallothionein (92.8 +/- 18.6 micrograms/g and 92.5 +/- 7.3 micrograms/g), but these did not exceed the level of the control group (94.5 +/- 8.4 micrograms/g) which had neither Cd nor Se injections. In the groups with testicular necrosis, concentrations of zinc (Zn) and magnesium (Mg) were decreased while an increase in concentrations of calcium (Ca) and iron (Fe) was observed. These results suggest that Se concentration must be maintained to prevent the testicular disorder caused by Cd. PMID- 10479890 TI - [An appraisal of the judgements of the Kumamoto Minamata Disease Certification Commission from May 1981 to July 1992 for applicants from December 1975 to April 1981]. AB - The differences of the diagnosis from December 1975 to April 1981 and from May 1981 to July 1992 of the Committee on Certification of Minamata Disease based on the medical records of 3,870 applicants who had been examined from December 1975 to April 1981 by the Committee were compared and analyzed from a statistical point of view. Three hundred forty inhabitants from December 1975 to April 1981 and two hundred ninety eight inhabitants from May 1981 to July 1992, in total six hundred and thirty eight inhabitants, were certified to have Minamata disease by the Committee on Certification of Minamata Disease. One hundred and eleven inhabitants who were judged likely to have Minamata disease by application of the present criteria presented by the Japanese Environmental Agency in 1977 were certified by the committee from May 1981 to July 1992, and one hundred and thirty three inhabitants who were judged not likely to have Minamata disease by application of the present criteria were certified from May 1981 to July 1992. The author concluded that certification of the patients of Minamata disease by the Committee was inconsistent with the results of applying the present criteria to the data and that the prevalence of the symptoms of the inhabitants certified from December 1975 to April 1981 was inconsistent with that from May 1981 to July 1992. PMID- 10479891 TI - [Surgical removal of subfoveal choroid neovascularization in senile macular degeneration]. AB - The follow-up of central scotomas and fixation--next to visual acuity--are important parameters for the evaluation of new therapies in AMD. PATIENTS AND METHODS: Twenty-three patients (age 67 to 91 years) with subfoveal CNV had SLO fundus-controlled perimetry before and 6-8 weeks after surgical removal of the CNV. The size and location of deep (0 dB) and relative (12 dB) scotomas were measured. Stability and location of fixation were analyzed. RESULTS: Fifty-six percent of patients gained (10% lost) more than 2 lines of VA; 52% of deep scotomas decreased in size (26% increased). No relative scotoma increased, but 63% decreased, some remarkably. Most scotomas had steeper borders postoperatively. Five of 7 patients were able to fixate again. Fixation moved slightly more peripheral in 4 patients and was otherwise unchanged. None of 7 patients whose fixation was close to their fovea preoperatively lost that fixation. CONCLUSION: Subfoveal surgery may stabilize the course of subfoveal CNV in AMD at 6 weeks follow-up. In some patients the major benefit can be a reduction of relative scotoma due to reattachment of the retina. As the location of fixation changes little with surgery and is typically located within the area of relative scotoma, visual function can improve. PMID- 10479892 TI - [Oxygen saturation of retinal vessels. Studies for measuring with polarized light]. AB - The accuracy of the spectrometric measurement of the oxygen saturation in retinal vessels is limited by its signal-to-noise ratio. The aim of this study was to investigate the possibility of enhancement of the reflection signal by the use of polarized light. MATERIALS AND METHODS: The Jena ophthalmospectrometer was equipped with two polarizing filters: one in the illumination and the other in front of the detector. Reflection spectra of erythrocytes streaming through a cuvette in the focus of an artificial eye were recorded. The influence of the polarization on the reflection spectra was investigated by rotating the polarizer in front of the detector. Furthermore, the degree of polarization of the light reflected from retinal vessels in vivo was determined. RESULTS: The degree of polarization of the light reflected from the erythrocytes was 0.6-0.8, whereas the polarization of light reflected by a standard white reflectance target was virtually zero. CONCLUSION: Polarized light can be used for the reduction of error in retinal vessel oximetry. PMID- 10479893 TI - [Sutureless corneal cataract surgery. Limbal release incisions for correcting astigmatism]. AB - BACKGROUND: To prevent surgically induced astigmatism following clear corneal cataract surgery. PATIENTS AND METHODS: Limbal relaxing incisions of 6 or 8 mm length respectively and 0.5 mm depth were performed in 52 patients with spheric cornea or astigmatism with the rule of 0.8 +/- 0.3 dpt after temporal corneal cataract incision. The same amount of patients, operated on with the same surgical procedure except keratotomies, served as control. RESULTS: After 6 months the with-the-wound-change (WTW) in the LRI axis using the Hollady method was -0.08 +/- 0.5 dpt in the spheric cases with limbal keratotomy and +0.5 +/- 0.7 dpt in the control cases. The patients with preoperative astigmatism with the rule showed a WTW in the LRI axis of -0.09 +/- 0.5 dpt in the keratotomy cases and +0.39 +/- 0.7 dpt in the corresponding control patients. CONCLUSION: Limbal relaxing incision is a reliable and safe procedure to reduce postoperative astigmatism after cataract surgery. PMID- 10479894 TI - [White-noise field campimetry in HIV+ patients]. AB - PURPOSE: The first morphological changes in eyes with HIV infection are microvascular disease of the retina with cotton-wool spots and microaneurysms. The study was performed to find out if evidence of disturbances of ocular microcirculation can be established by non-invasive methods. PATIENTS AND METHODS: Twenty-seven patients with HIV infection and without opportunistic infections underwent thorough ophthalmologic examination with threshold-oriented, suprathreshold perimetry (TAP 2000 ct, Oculus) and white-noise field campimetry (TEC, Oculus). RESULTS: Visual field examination was normal in 23 out of 27 patients (85%), whereas 4 patients showed relative field defects in at least one eye. In white-noise field campimetry 13 out of 23 perimetrically unaffected patients (56%) perceived scotomas in one or both eyes. These scotomas were not stable. Three of 4 patients with relative scotomas in the visual field had cotton wool spots in the retina and showed a stable scotoma in campimetry. Visual acuity, IOP, and cup/disc ratio were within normal ranges. CONCLUSION: White noise field campimetry complements the standard examination of patients with HIV and might be capable of indicating disturbances of ocular microcirculation by a non-invasive method before morphological changes in the retina can be seen. PMID- 10479895 TI - [Postoperative complications of Toti DCR (dacryocystorhinostomy. An indication for canalicular surgery]. AB - Failures with external DCR have also been attributed to canalicular occlusion. Since 1976 we have documented all patients with surgical reconstruction of occluded canaliculi. The indication, different surgical methods of choice and functional prognosis were evaluated. PATIENTS AND METHODS: From 1976 to 1997 we have treated 272 cases. The surgical technique canaliculodacryocystorhinostomy (n = 132) was performed in 45%. Thirty operations (11%) of occluded lacrimal canaliculi due to external DCR were done. The patients ranged in age from 2 to 79 years. We examined all 29 patients (10 male, 19 female) with 30 operations (1 patient was operated on in both eyes). RESULTS: The surgical procedure was performed after an interval of 1 month to 24 years (average 4 years). Seven times a second external DCR was done. Special causes of canalicular obstruction were severe previous soft tissue and bone trauma (4 patients) and congenital defects (3 patients). Canaliculorhinostomy was performed 15 times, canaliculodacryocystorhinostomy 14 times and conjunctivorhinostomy once. Fifteen operations had good functional results (without any symptoms). Five patients had good irrigation, but a prolonged fluorescein dye disappearance test. There were 10 failures with complete closure of the canaliculi. CONCLUSION: If failure with external DCR is caused by canalicular occlusion, the microsurgical technique has a possible success rate for long-term results of about 50%. Before hand, however, all prophylactic measures have to be taken to avoid this postoperative failure. PMID- 10479897 TI - [Long-term stability of astigmatism outcome after arcuate lamellar keratotomy. 3 year results of a prospective study]. AB - INTRODUCTION: The correction of low to moderate astigmatism is possible today by means of photoablation (PRK), while the treatment of moderate and higher astigmatism still involves refractive keratotomy. Experience has shown that cataract surgery, using modern tunnel techniques with self-healing incisions, results in earlier stability in both the refractive outcome and wound healing. In this study, we attempted to combine the advantages of lamellar keratotomy with those of a pair-wise T-incision as arcuate lamellar keratotomy (ALK). PATIENTS AND METHODS: The clinical outcome of 41 patients who underwent ALK was investigate in a prospective study over a period of 3 years. The pre- and postoperative investigations undertaken included the measurement of astigmatism using a Zeiss keratometer, uncorrected visual acuity, and corrected glare vision using a Humphrey autorefractor. All patients had astigmatism between 2.0 and 7.0 D. Together with a uniform arcuate incision, we used 7 mm (n = 26) and 8 mm (n = 15) mm zones for correction. RESULTS: The average preoperative astigmatism was 4.01 +/- 1.90 (median, 3.50) D. After a 3-year follow-up the average postoperative astigmatism was 1.59 +/- 1.29 (median, 1.38) D. The astigmatic change induced (Jaffe) after 3 years was about 3.23 +/- 2.23 (median, 3.16) D. The average uncorrected visual acuity (log MAR) before ALK was 0.20 +/- 0.12 (median, 0.22) and after follow-up, 0.41 +/- 0.14 (median, 0.39). Corrected glare vision before surgery was 0.23 +/- 0.19 (median, 0.10) and afterwards, 0.25 +/- 0.22 (median, 0.14). CONCLUSIONS: Arcuate lamellar keratomy (ALK) stood the test as a routine clinical procedure for correction of moderate astigmatism with stable postoperative functional outcomes. We did not find impairment of glare vision following this procedure. PMID- 10479896 TI - [Etiology of corneal opacities after plasminogen activator-induced fibrinolysis of the anterior chamber]. AB - BACKGROUND: After treatment of anterior chamber fibrinous reactions by tissue plasminogen activator (TPA), irreversible corneal opacifications (calcium phosphate) have been observed. To understand the mechanism of these opacifications an animal model was developed. MATERIAL AND METHODS: In rabbits the lens was removed by phacoemulsification. The surgical procedure was completed by an injection of TPA (25 micrograms) into the anterior chamber. In a second group TPA fibrinolysis (25 micrograms) was induced 10 min after injection of autologous blood. In a third group 25 micrograms of TPA was injected into the anterior chamber after circumscribed mechanical lesion of the corneal endothelium. Changes in corneal structure and transparency were determined by biomicroscopy and histopathologic examination. RESULTS: After lensectomy or mechanically induced lesion of the corneal endothelium followed by TPA injection, sharply defined interpalpebral corneal opacifications developed within 3 to 8 days. Histologically, deposits were located in Bowman's membrane and in superficial stromal layers. No opacifications developed after fibrinolysis of an intracameral clot. CONCLUSIONS: Corneal opacifications as seen in humans after fibrinolysis by intracameral injection of TPA requires a temporary disturbance of the endothelial function. This allows phosphate (buffer of TPA) and calcium (aqueous humour) to distribute within the corneal stroma. Then there are insoluble calcium phosphate precipitates because of recovery of the endothelial function and dehydration of the cornea. PMID- 10479898 TI - [Bacterial keratitis. Microbiological analysis as a principle for therapeutic recommendations]. AB - PURPOSE: The study presented differentiates between the aetiological agents of bacterial keratitis in patients with and without a history of contact lens wear. Based on these results, recommendations are given for optimal antibiotic primary therapy. PATIENTS AND METHODS: Swabs and corneal scrapings were taken from 218 patients referred to the University Eye Hospital in Munich with a diagnosis of bacterial keratitis from 1989 to 1997. Ninety-two of these patients had a history of contact lens wear; 126 had none. The germs were isolated and identified by staining and microscopy. Observing polymicrobial growth in 51 patients, a total of 275 germs was isolated. RESULTS: The most frequent pathogens were Staphylococcus epidermidis (44%), S. aureus (18%), Streptococcus spp. (10%), Propionibacterium acnes (7%) and Pseudomonas aeruginosa (6%). Gram-negative germs were nearly exclusively isolated from contact lens wearers, gram-positive germs were predominant in non-contact lens wearers. CONCLUSION: Keratitis in patients with a history of contact lens wear is often caused by aggressive gram-negative germs. Those cases should immediately be treated with quinolones and erythromycin. In keratitis caused by gram-positive pathogens, a combination with aminoglycosides and erythromycin is sufficient. PMID- 10479899 TI - [Difficulties of fungus detection in corneal culture medium]. AB - It is not always possible to prevent the growth of microorganisms in organ culture for cornea preservation, despite many prophylactic measures. It is especially difficult to prove the presence of fungi in the cultural medium. MATERIALS AND METHODS: A culture medium was examined for sterility after 8 days' storage of cornea in organ culture. To prove the presence of fungi a culture of Sabouraud 2% glucose-agar was prepared and its growth examined by light microscopy. RESULTS: After 8 days of preservation we noticed a color change in the cultural medium and suspected contamination with fungi. Coagulase-negative Staphylococci could be cultivated from the conjunctival smear obtained before preparation of the cornea only. Routine screening of microbiological contamination did not show any results. We were able to identify an Aspergillus species only after preparing a special culture. The conjunctival smear as well as the cultural medium of the other eye of the same donor showed no contamination. CONCLUSIONS: In spite of the fact that microbiological contamination can be seen macroscopically, it is difficult to prove the presence of a specific microorganism and even more so when dealing with fungus. Especially in these cases the incubation of the cornea in media might have an advantage because contamination can be suspected by just looking at the medium. By excluding these preparations from transplantation we can possibly prevent infections, even when routine examinations show negative results. PMID- 10479900 TI - [Cystic conjunctival tumor with hemorrhage. Lymphangiectasia haemorrhagica conjunctivae Leber (lymphangioma of the conjunctiva)]. PMID- 10479901 TI - [Ophthalmologic periodicals and publications on the internet]. PMID- 10479902 TI - [Pathogenesis, clinical aspects and therapy of eye involvement in allergy]. PMID- 10479903 TI - Utility and complications of permanent venous access devices (PVAD) in oncological treatments. Follow-up of 100 cases. AB - BACKGROUND: Permanent venous access devices (PVAD) are nowadays routinely implanted and used with some morbidity for the oncological treatments. The adequate timing of implantation based on the number of treatments, the survival rate and the complications has not yet been well estimated. METHODS: A hundred permanent venous access devices placed in oncological patients were followed-up prospectively. RESULTS: No mortality was seen due to the surgical act. A 11% morbidity rate was noted, largely due to infections, with 6 patients needing a second surgery. On average, 6 chemotherapy cycles were done after placing of the permanent venous access device. CONCLUSIONS: Due to these results and an average survival rate of 10.7 months, we suggest the placing of a permanent venous access device early in the management of oncological patients requiring chemotherapy cycles, so as to increase the comfort of the patient and to safeguard his peripheral venous system. PMID- 10479904 TI - The absence of correlation between immunoregulatory T cells and induced lymphoproliferative response in treated B-chronic lymphocytic leukemia patients. AB - BACKGROUND: Many data suggest T cell functional impairment in B-cell chronic lymphocytic leukemia (B-CLL). The mechanism responsible for this phenomenon is still unresolved. METHODS: In 88 B-CLL patients (RAI II-IV) the relationship between immunoregulatory T cells and PHA induced lymphoproliferative response (LPR) was analysed before and after the therapy. The number of peripheral blood CD3+, CD4+ and CD8+ T lymphocytes was determined by indirect immunofluorescence assay using monoclonal antibodies. LPR was estimated in whole blood culture method. RESULTS: The absolute number of CD3+, CD4+ and CD8+ cells in untreated CLL patients was much higher than in healthy controls (n = 26), but the percentages of these subpopulations, CD4/CD8 ratio and LPR to PHA were significantly (p < 0.00001) decreased. The chemotherapy induced a significant rise of CD3+ and CD4+ percentages (p < 0.006 < p < 0.022 respectively) in comparison to baseline levels, but their levels remained significantly (p < 0.00001) lower than the controls. The CD4/CD8 ratio was also elevated after the therapy (p < 0.048) but remained below the normal value as well. The absolute number of CD3+ and CD4+ T cells were normalized after treatment, while the CD8+ cells were still higher (p < 0.044) than controls. The increase of LPR has been registered after treatment, but it failed to reach the control values. We could not find any correlation between the number of immunoregulatory T cells and induced LPR (r = 0.07, for CD4+; r = 0.09 for CD8+ cells). CONCLUSIONS: These data indicate some profound lymphoid cell defect in CLL patients affecting CD8+ proliferation as well as LPR. PMID- 10479905 TI - Stability of serum TSH concentration after non refrigerated storage. AB - BACKGROUND: The ignorance of how sensitive serum samples for defined assays may be, can lead to overprotecting measures. This attitude not only increases cost and labor in everyday routine but may be responsible for not evaluating series of measurements if performed in serum samples not well stored. METHODS: Thyrotropin (TSH) concentration in serum was tested in three pooled samples drawn from various patients, before and after four days storage at freezing (-15 degrees C), low (4 degrees C), and room temperature (18-22 degrees C). TSH concentration was measured in each part, in 22 replicates using a commercial assay. RESULTS: The variation of the measured values due to the storage conditions was less than 10%, not exceeding the accepted between assay variation. CONCLUSIONS: For the estimation of TSH concentration, patient serum samples may be stored up to four days at 4 degrees C or even at room temperature without significant loss of measurement accuracy. PMID- 10479906 TI - The influence of furosemide on free water clearance. AB - BACKGROUND: Laboratory measurements of osmolality and electrolyte concentrations are useful as a source of clinical and pathophysiologic information on kidney function. We obtained different conditions of baseline diuresis in 54 rats, which were then treated with furosemide 10 mg/kg. From measurements of plasma (P) and urine (U) osmolality and Na, we calculated the free water clearance (CH2O) and the contributions of Na+ and non-Na(+)-solutes to its changes during diuretic administration. RESULTS: The results show that furosemide abolishes both urine diluting and concentrating ability, by reducing the contribution of non-Na(+) solutes to Uosm and the formation of CH2O, while the contribution of Na+ to Posm remains unchanged. During furosemide the urines approach isosmoticity irrespective of the baseline Uosm with an asymptomatic function similar to that imposed by osmotic diuresis. It is suggested that the overflow to the concentrating sites of the nephron overwhelms their normal transport capacity independently of their baseline water permeability. The disruption of the transepithelial osmotic gradient caused by the drug impairs the transepithelial osmotic flow, leading to the excretion of isosmotic urines. CONCLUSIONS: The effect of non-Na(+)-solutes in the laboratory measurement of urine constituents can be reduced to that of plasma determination by diluting the urines according to empiric formulas derived from the present data. PMID- 10479907 TI - Biohumoral parameters and bone mineral content in the identification of high risk subjects for postmenopausal osteoporosis. AB - BACKGROUND: The aim of this study was to evaluate the possibility of identifying women with a high risk of postmenopausal osteoporosis by using computerised bone mineral analysis (CBMA) associated with markers of bone turnover in order to provide a valid and reliable screening test. METHODS: A total of 925 patients were evaluated, 252 of whom had already undergone a preliminary densitometric test six months earlier and were diagnosed as fast bone losers. 225 of them (89%) also showed altered bone turnover markers. CBMA was negative in the remaining 673 patients but 13 patients showed altered bone turnover markers and three of the latter then showed a positive CBMA 18 months later. The 673 patients who, after six months of study, were not fast bone losers were monitored over time. RESULTS: It emerges from these results that fast bone losers are characterised by higher levels of hydroxyprolinuria and calcium, lower levels of oestrone and estradiol, and reduced body weight compared to healthy subjects. CONCLUSIONS: This simplified method enabled 79% of the fast bone losers to be identified, whereas densitometry identified 87.5% of the high-risk subjects. The main advantage of our simplified method compared to the measurement of bone mineral content is that it identifies the majority of fast bone losers in the initial postmenopausal period, before a substantial reduction in BMC has taken place. PMID- 10479908 TI - Measurement of specific IgEs in the diagnosis of drug allergy. AB - BACKGROUND: The anamnesis and, in certain cases, skin tests are indispensable methods for the correct diagnosis of allergy to drugs. However, in some cases, these two techniques alone, prove insufficient in achieving reliable predictive elements for drug allergies. Besides, the use of skin testing is often discouraged, due to the potential risks involved. METHODS: In this study, the diagnostic efficiency of the measurement of specific IgE for a selected group of 13 drugs was evaluated. The test was carried out on 96 patients showing clinical signs directly attributable to the administration of one particular drug (in 38 of the 96 cases) or more drugs (58/96 cases). The symptoms appeared after a week or more from the beginning of the drug treatment; this time interval is compatible with the development of an immune response. RESULTS: The drugs most frequently causing adverse reactions were salicylic acid, noramidopyrin, propiphenazone, other non-steroidal anti-inflammatory drugs (NSAIDs), trimetoprim, antibiotics. RAST was applied to 13 drugs and was positive for at least one of the drugs tested in 31/96 patients. CONCLUSIONS: The results of the study confirm the validity of the RAST technique as a useful element in improving and making the overall diagnosis of allergies to drugs more reliable. PMID- 10479910 TI - Plasma thrombomodulin levels in lung cancer patients. AB - BACKGROUND: Thrombomodulin (TM) is a glycoprotein and besides its anticoagulant property it is accepted as an onco developmental antigen. Considering these properties we hypothesised that active TMB might have a role in cancer cell behaviour. METHODS: We measured serum TMB levels by the enzyme immunoassay method in 40 patients with lung cancer and 20 healthy subjects. RESULTS: TMB levels were found as 54.6 +/- 11.5 in controls and 60.9 +/- 31.2 ng/ml in lung cancer patients TM levels were 59.4 +/- 24.2 ng/ml in 24 epidermoid carcinoma patients, 65 +/- 50.2 ng/ml in 10 small cell carcinoma patients and 60 +/- 18.4 ng/ml in 6 adenocarcinoma patients. Patients with stage 4 and distant metastasis had a value of 65 +/- 33.9 ng/ml. No significant differences were found between healthy subjects and cancer patients. Also the differences between 3 groups of patients were insignificant. CONCLUSIONS: It was concluded that serum TMB levels could not be accepted as a tumour marker in lung cancer patients. PMID- 10479909 TI - Tubular reabsorption and sodium excretion during urine reinfusion. AB - BACKGROUND: The mechanisms responsible for the natriuresis that follows urine reinfusion was investigated in rats by clearance and micropuncture techniques. METHODS: In each animal two urine reinfusion periods (R1 and R2) were performed and compared to a non-urine-reinfusion, saline infusion period (S) sandwiched between them. RESULTS: Switching from urine reinfusion to an equivalent rate of saline loading was followed by a fall in Na excretion from 1.9 +/- 0.5 to 0.5 +/- 0.2% of filtered load, p < 0.002. Urine osmolality rose, and urine to plasma inulin concentration ratio rose significantly from 73 +/- 14 to 147 +/- 21 (p < 0.002). The changes in GFR, SNGFR, absolute and percent proximal reabsorption could not account for these findings. A reduced Na excretion coupled to increased urine osmolality indicates enhanced transport along a segment responsible for the urinary concentrating mechanism. Thus the data can be interpreted then as due to enhanced reabsorption along the ascending limb of Henle's loop. These changes were reversed by reinstituting urine reinfusion after the S period. The consensual changes in Na+ and K+ excretion excluded an effect of urine reinfusion on the distal exchange site. There was a continuous fall in proximal reabsorption from R1 (76 +/- 3%) to S (69 +/- 3%) to R2 (62 +/- 5%) which was inversely correlated with the changes in hematocrit (R = 0.49, p < 0.026). This indicates that part of the late diuresis and natriuresis was due to volume expansion. An osmotic effect of reinfused urine solutes was suggested by a late rise in plasma osmolality, from 312 +/- 13 to 323 +/- 8 mOsm/kg. Osmotic diuresis could have exerted additive effects upon those of volume expansion, accounting for the late fall in proximal reabsorption. CONCLUSIONS: We conclude that the acute effects of urine reinfusion are due to changes in transport of solutes and permeability to water along distal tubular segments. The changes in plasma osmolality during the last period of the present acute experiments, suggest the possibility that solute retention may be linked to the chronic effects of urine reinfusion. PMID- 10479911 TI - Radio- and chemotherapy variably affect the general immunocompetence of lung cancer patients. AB - OBJECTIVE: The evaluation of the effects of radiotherapy and chemotherapy on the immune status of lung cancer patients. EXPERIMENTAL DESIGN: Prospective nonrandomized study. SETTING: Hospitalized care. PATIENTS: 121 patients with unresectable non-small-cell lung cancer (Stage IIIb or IV), who were planned for radiotherapy (n = 81) or chemotherapy (n = 40). MEASURES: The relative and absolute numbers of blood T lymphocytes and monocytes, as well as the mitogen induced proliferative response of the former, and phagocyting capacity of the latter cell subpopulation, were determined in patients before starting any therapy. In radiotherapy (RT)-treated group, the immune parameters were evaluated after 45 Gy and 60 Gy had been given. In chemotherapy (ChT)-treated group, the same parameters were determined three weeks after the 2nd and 4th cycle of ChT. RESULTS: The number and proliferative response of T lymphocytes were significantly (p < 0.001) lower, while the number and phagocyting capacity of monocytes were significantly (p < 0.001) higher in all patients before therapy, in comparison to the controls. After RT, the T cell number and proliferative response were significantly (p < 0.001) decreased, while the number of monocytes and their phagocyting capacity remained unchanged, when compared to the pretreatment values. Unlike RT, chemotherapy did not change any investigated parameter, except for the phagocyting activity of monocytes, which was significantly (p < 0.02) decreased, in comparison to the pretreatment value, after four cycles of ChT only. CONCLUSIONS: Two cancer treatment modalities- radio- and chemotherapy--variably affect the immune status of lung cancer patients. The initial great disturbances of general immunity parameters are further aggravated by radiotherapy. Unlike RT, chemotherapy exerts no suppression at all; on the contrary, it tends to normalize some of the parameters of cellular immunity of lung cancer patients. PMID- 10479912 TI - Onset of myasthenia gravis in a patient affected by rheumatoid arthritis never treated with disease-modifying anti-rheumatic drugs. AB - We describe a patient affected by rheumatoid arthritis (RA) that developed myasthenia gravis (MG) after 20 years of illness. The peculiarity of this case concerns both the rare association between these diseases and the fact that the patients had never assumed disease modifying antirheumatic drugs. These treatments have been associated in some clinical reports with the onset of MG during the clinical course of RA. To our knowledge this is the first case described in medical literature up to now. PMID- 10479913 TI - Endocrine changes in postmenopausal women after high-dose danazol therapy. AB - BACKGROUND: Purpose of the study was to investigate changes in the hormonal pattern in postmenopausal women after high dose danazol therapy. In 16 postmenopausal women (mean age 66.4 +/- 8.6) treated with danazol 1.2 g/die x21 we studied serum levels, before and after therapy, of FSH, LH, E2, fE2, T, fT, D4 A, DHEAS, SHBG. METHODS: FSH, LH, SHBG by IRMA kits (DPC); E2, T, DHEAS, D4-A by RIA kits (Sorin Biomedica) after celite microcolumn chromatography; fT by coat-A count kit (DPC); fE2 by equilibrium dialysis; serum albumin by bromocresol-green colorimetric methods. RESULTS: After therapy there was a statistically significant decrease of LH by 13%, of FSH by 19%, SHBG by 84% and increase of fE2 by 24.5%, fT by 77%. Other hormones show changes that are not statistically significant. CONCLUSIONS: In postmenopausal women danozol therapy, even if at a higher dose, has about same effect on hormonal pattern as in normally cycling women. Danazol showed a clear antigonadotropic effect and can play a role in therapy when reduction of gonadotrophins and E2 secretion is desirable. PMID- 10479914 TI - Clinical validation of an automatic device measuring blood pressure in the fingers. AB - BACKGROUND: To assess the agreement between a new automatic device (FS-20D) using a cuff-oscillometric method to measure arterial blood pressure (BP) in the fingers and a standard mercury sphygmomanometer. METHODS: The blood pressure measurements were taken in a sequential order, in a sample of both normotensive subjects (n. 57) and slight to moderate hypertensive patients (n. 28) without vascular complications. RESULTS: The mean sphygmomanometer-monitor difference was 0.52 +/- 4.57 mmHg for systolic and 0.25 +/- 4.41 mmHg for diastolic values; the agreement limits were: SBP -8.6 divided by 9.6 mmHg, 95% CI: -0.5 divided by 1.5; DBP: -8.6 divided by 9.1 mmHg, 95% CI: -0.7 divided by 1.2. The grade of agreement between the monitor and the sphygmomanometer was "A" (British Hypertension Society) for both systolic and diastolic values (difference of readings < 5 mmHg: 82%; < 10 mmHg: 97% for systolic blood pressure, 98% of diastolic blood pressure). CONCLUSIONS: The monitor was proved to be reliable with a good level of precision and accuracy. The FS-20D monitor may be used in self-monitoring of blood pressure of patients with slight to moderate hypertension. PMID- 10479915 TI - Fungi of Candida species in the pharynx and some parameters of humoral and cellular immunity in patients. AB - BACKGROUND: The pharynx is one of the open spaces where fungi can settle easily. The purpose of the investigation was to estimate an overall prevalence of fungal colonization and some parameters of humoral and cell immunity. METHODS: The study consisted of 167 patients, from whom swabs of the posterior and lateral part of the pharynx were taken for mycological examinations, during which tests based on morphological and biochemical (zymogram, auxanogram) characteristics were used. In all cases, the leukocyte count, overall protein level, and protein individual fractions were determined. Also determined were the levels of IgA, IgM and IgG immunoglobulin in blood serum, and IgA immunoglobulin (S-IgA) in saliva. At the same time, blastic transformation test and rosette tests: E (ER) to determine lymphocytes T count, as well as EA (EAR) and EAC (EACR) to detect lymphocytes B, were performed. RESULTS: Fungal infection was found in 2/3 of patients. Fungi were classified as belonging to six species of Candida, which are considered to be important etiological factors of mycoses; the most common fungus was Candida albicans (in 86 cases). All cases of candidosis showed a decreased count of lymphocytes T, with their function deficient, and most of them a low S-IgA level in saliva (over three times lower than the norm) and high mean values for sedimentation rate. CONCLUSIONS: In patients in our study the IgA level in serum remained within normal limits, yet the ratio fo this fraction to other immunoglobulins deviated from that in healthy individuals. PMID- 10479916 TI - Panic disorder in children and adolescents. AB - Panic disorder (PD) is a well-known and frequently described psychiatric disorder in adults, that can cause clinically significant distress and impairment of social and occupational functioning. It is characterized by a discrete period of intense fear and discomfort, that develops abruptly and reaches a peak in 10 minutes or less, with other somatic and cognitive symptoms. PD in prepubertal children and early adolescents in rarely reported, but 18% of adult patients with PD indicate onset of PA before 10 years of age. Probably many of the prepubertal cases are being misdiagnosed, and/or they can have a different clinical expression from adults. The lack of identification of these affected subjects can have serious consequences on social and academic development. The aim of this paper is to review clinical literature describing PD in children and adolescents. Prevalence, specificity of clinical features, comorbidity, instruments for diagnosis are described. Directions in pharmacological, psychotherapeutic and educational management of PD in children and adolescents are suggested. PMID- 10479917 TI - Acromegaly and intestinal neoplasms. AB - Acromegalic subjects show increased frequency of neoplastic lesions in the colon and rectum with respect to the general population. Recent prospective studies using colonoscopy have shown a 3 time higher prevalence of intestinal polyps and up to 4 time increased presence of colorectal cancer in acromegaly, independently of sex, age, duration of disease and clinical status of the patients. The polyps are distributed throughout the extension of the large bowel and are often multiple, showing at least two different histologic types: hyperplastic and adenomatous. Sometimes they are associated with intestinal carcinomas. Pancolonoscopy is the procedure of choice for the diagnosis of large bowel neoplasms, even though it may be difficult to complete in these subjects because of the frequent presence of an enlarged and elongated colon. It shows a higher sensitivity and specificity compared to other tests such as the barium enema, fecal occult blood test and serum levels of carcinoembryonic antigen. Therefore, it is recommended to follow up acromegalic patients using pancolonoscopy to obtain early detection of neoplastic lesions in the large bowel. PMID- 10479918 TI - Non specific aorto-arteritis. Clinical picture, diagnosis and therapy. AB - This study is aimed at analyzing clinical features, angiographic findings and evolution of Takayasu's arteritis and the criteria adopted to establish the indication for non-surgical versus operative treatment. Eighteen patients affected by non specific aortarteritis were observed and treated at our Department between 1973 and 1996. All patients met the American College of Rheumatology 1990 criteria of classification of Takayasu's arteritis. Sixteen patients were young females. Two patients were males. Nine patients underwent surgical procedures. One young female underwent a PTA. All eight asymptomatic patients were only medically treated. One patient died some days after an aorto bicarotid bypass graft due to acute myocardial infarction. Two other patients died two and four years after intervention for renal and cardiac failure respectively. One patient after an aorto bicarotid bypass underwent a left hemiplegia due to thrombosis of the right graft branch. All the other 14 patients either surgically or medically treated are well and are under strict surveillance through rigorous follow-up. The 10 patients who underwent surgical or interventional radiological treatment were certainly the most seriously affected patients and were symptomatic (presented neurological disturbances or a severe hypertensive state). This fact explains, to some extent, the mortality and morbidity rate observed in this group. The seven medically treated patients were completely asymptomatic in spite of a major involvement of various vascular districts. PMID- 10479919 TI - Pneumococcal arthritis in the wrist and ankle. A case report and short review of the literature. AB - In this paper we report a 48-year-old man with septic arthritis of the wrist and ankle due to Streptococcus pneumoniae. No known predisposing factor was found. Only about 300 cases of pneumococcal arthritis have been reported in the literature since 1888 and less than 10% of these affect the wrist. The management of bacterial arthritis is reviewed. PMID- 10479920 TI - A case of lung myelolipomatosis in a patient with bronchial carcinoid. AB - Myelolipomas are very rare benign tumours composed of an admixture of mature adipose tissue and normal haematopoietic cells. Although they are most commonly found in the adrenal glands, extra-adrenal myelolipomas are documented. We described a case of myelolipoma arising in the lung in a 52-year-old man. The lesion was found incidentally in association with a carcinoid. To our knowledge, this is the second instance of this neoplasm presenting as a lung lesion, and the first case associated with bronchial carcinoid. Pathogenesis and aetiology of myelolipomas are referred to in this paper with special regard to the clinical and pathological findings. PMID- 10479921 TI - [BCG vaccination: characteristics of drugs and causes of vaccinal complications]. AB - Series of BCG and BCG-M vaccines causing complications in children vaccinated in Russia's areas in 1995 were checked up. All the series met the Russian and international specifications for BCG and BCG-M vaccines. The incidence of postvaccinal lymphadenitis was 5 times less than the limit laid down by an official article. The properties of the agents produced no significant effect on the number and type of postvaccinal complications. The risk for complications was much less when BCG-M was used. The risk factors were defects of intradermal vaccine administration and contaminant abnormalities prior to vaccination and at the moment of local vaccinal response. PMID- 10479922 TI - [Mental evaluation of new cases of pulmonary tuberculosis in different treatment settings]. AB - New one hundred and five cases of pulmonary tuberculosis were examined to evaluate the mental status in different treatment settings: a day hospital, a polyclinic, and a hospital. The Aizenok questionnaire. Shihan's test, and a clinical and symptomatological questionnaire and interviews with patients were used for psychological studies. In the day hospital and polyclinic, therapy improved the mental status of the patients at discharge. They showed their adequate attitude towards their disease, felt well and had great visas for their future social and familial life. They all were eager to resume their former work, attempted to preserve their social and working status and 16 (72.7%) patients treated at the day hospital and 18 (75%) outpatients continued working. PMID- 10479923 TI - [Psychosomatic relationships in sarcoidosis]. AB - The procedure developed by the Bekhterev Leningrad Research Institute was used to define the attitude towards the disease in 40 patients with sarcoidosis. A harmonic attitude towards the disease was detected only in 20% of cases. There was a relationship between the magnitude of clinical manifestations of sarcoidosis and the type of an attitude towards the disease. Hyperneurotization was observed in 50% of female patients with sarcoidosis. The study showed it necessary to perform psychotherapy for sarcoidosis on an individual basis to improve their psychoemotional status and to reach a better mutual understanding between the patient and the medical staff. PMID- 10479924 TI - [Analysis of mortality rates among patients with pulmonary tuberculosis]. AB - The outpatient medical cards and history cases of those who died, autopsy protocols of 428 dead patients with various forms of tuberculosis were analyzed. Of them, 86 patients had been followed up less than a year. The patients treated with antituberculous drugs died not only from a progressive specific process, but from nonspecific complications. The factors predisposing to death were late detection of tuberculosis and irregular treatment, alcohol and drug abuse. PMID- 10479925 TI - [Radionuclide diagnosis of capillary pulmonary blood flow in new cases of pulmonary tuberculosis]. AB - The radionuclide technique pulmonary perfusion scintigraphy was used to study 124 new cases of pulmonary tuberculosis before therapy and 25 controls. The data indicated that perfusion impairments were significant and rather noticeable in 84% of new cases of the disease. Evidence was provided for that pulmonary capillary blood flow was associated with the duration of the disease, with the initial type and phase of a tuberculous process, and with clinical manifestations. There was a predominance of significant and drastically marked vascular lesions in patients with infiltrative and disseminated tuberculosis under the present conditions. PMID- 10479926 TI - [X-ray computed tomography in the diagnosis of pulmonary tuberculosis]. AB - The paper provides a detailed analysis of 120 cases of pulmonary tuberculosis that was mainly detected by X-ray computed tomography (XCT). The study was performed on third-generation XCT plants. The authors highly appreciate the contribution of XCT to the diagnosis of tuberculosis with emphasis on some organizational issues. They suggest that third-forth-generation XCT plants that show its high effectiveness in detecting major lung diseases, including tuberculosis, and cost-efficiency should be widely introduced into the outpatient setting. PMID- 10479927 TI - [Indications for surgical treatment of chronic and encysted pleurisy]. AB - If exudative pleurisy developed unfavourably with the formation of encystment, the development of massive pleural impositions, the deterioration of cardiorespiratory functional parameters, reparative operations, such as pleurectomy with decortication, were performed at month 3-4 of the onset of the disease with the minimum number of intra- and postoperative complications, with the more rapid and proper anatomic and functional recovery. Postoperative complications occurred in 75% of patients at months 1-2 of the disease, in 25-30% at months 5-12. PMID- 10479928 TI - [Use of native and lyophilized Loewenstein-Jensen's solid media in determining drug resistance on mycobacterium tuberculosis strains]. AB - The results of determination of drug medium depend on both individual cultural properties of the strains tested and the type of the medium used. The composition of a medium is of significance for some strains: the latter grow in any medium and are characterized by the same spectrum of resistance. However, most drug resistant strains have higher feeding demands which are not fully met on the lyophilized Lowenstein-Jensen medium. The strains in the lyophilized medium show an incomplete spectrum of resistance and less grow than do those in freshly prepared Lowenstein-Yersen medium. PMID- 10479929 TI - [Effects of sodium nucleinate on immunological and nonspecific responsiveness in elderly and senile patients]. AB - Sixty-two elderly and senile patients with pulmonary tuberculosis were examined. Of them 32 had multimodality therapy involving sodium nucleinate. The inclusion of sodium nucleinate into a complex of therapeutical measures for the patients promotes improvement of T-cellular immunity. The patients taking sodium nucleinate were also found to have enhanced specific immunity against tuberculosis. In these patients, stimulation of immunity and nonspecific responsiveness is accompanied by the higher clinical benefits of treatment, as shown by shorter periods in abacillation, in the cessation of signs of intoxication, and in the resolution of infiltrative events and in the closure of decay cavities. PMID- 10479930 TI - [Value of scanning electron microscopy in evaluation of peripheral red blood cells shape in patients with pulmonary tuberculosis for treatment effectiveness evaluation]. AB - The morphology of red blood cells in patients with pulmonary tuberculosis was studied by scanning electron microscopy during enterosorption. The use of an enterosorbent caused a significant decrease in the count of abnormally changed cell elements and an increase in normocytes, which suggests that detoxification has a stimulating effect. Enterosorption promotes less content of altered peripheral red blood cells, which leads to improvement of its cellular composition in the presence of clinical manifestations of the decreased rate of a tuberculous process in the lung and which is suggestive of the detoxifying impact of this treatment. PMID- 10479931 TI - [Changes in the production of interleukin-1 beta, tumor necrosis factor-alpha and interleukin-2 in relation to the immunity in patients with pulmonary tuberculosis]. AB - The specific features of production of IL-1 beta, TNF-alpha, IL-2 were studied in 74 patients with various forms of tuberculosis by taking into account the magnitude of an immunological response. Tuberculin, phytohemagglutinin, prodigiosine were used as inducers of the synthesis of cytokines. Heterodirection was found in the changes of elaboration of cytokines in similar immunological disorders in persons with different clinical forms of tuberculosis. Examination of patients with infiltrative tuberculosis indicated that the increased synthesis of TNF-alpha and IL-2 was to a greater extent associated with the activation of cell-mediated immunity and that of IL-1 beta with its inhibition. The relationships found between the production of cytokines and IgA and IgM levels are suggestive of their involvement in the regulation of immunoglobulin synthesis. Cytokine spectral alterations are associated with the changes in specific lymphocytic populations. In patients with infiltrative tuberculosis, the production of TNF-alpha and IL-1 beta was directly related to the proportion of CD4+ and CDS8+ and that of IL-2 is associated with the proportion of CD25+ and CD20+ and with the count of lymphocytes. At tuberculin stimulation of mononuclear cells, it is expedient to bear in mind the detection rate of cytokines and the level of their production. It was shown that the measurement of IL-1 beta, TNF alpha, IL-2 may be used in the treatment of tuberculosis to assess the patients' immunological response and in the choice of immunomodulating therapy. PMID- 10479932 TI - [Role of neutrophilic leukocytes in pathogenesis of idiopathic fibrosing pulmonary alveolitis]. AB - The study was undertaken to obtain additional data on the role of neutrophils in the pathogenesis of idiopathic pulmonary fibrosis (IPF). Various functional parameters of neutrophilic functional activity, i.e. phagocytosis, chemotaxis, random migration, oxidative metabolism (3 types of chemiluminescence: spontaneous formylmethionyl peptide-stimulated, and prodigiosan-stimulated), and the level of circulating immune complexes of different size were determined in patients with IPF and healthy individuals. The patients showed a decrease in phagocytosis and an increase in neutrophilic chemotaxis and oxidative metabolism, which was related to the output of toxic oxygen forms. The changes revealed were found to be associated with the activity of an pathological process. The findings suggest that neutrophils can be involved both in the development of and in the occurrence of a pathological process in IPF. PMID- 10479933 TI - [Breast sarcoidosis]. PMID- 10479934 TI - [Pathomorphology of laryngeal tuberculosis. (from 50 years of own observations)]. PMID- 10479936 TI - [Smoking as a risk factor for chronic bronchitis (review of literature)]. PMID- 10479935 TI - [Effectiveness of superhigh-frequency electromagnetic radiation in multimodal therapy of patients with pulmonary tuberculosis]. PMID- 10479937 TI - [Chronic obstructive lung disease]. PMID- 10479938 TI - [Ambulatory follow-up of patients with respiratory tract tuberculosis and ways of its improvement]. AB - Analysis of registry groups I and II patient cohorts indicated that to follow up the patients in the active tuberculosis groups were unpracticable. Based on the their own findings and the WHO recommendations on treatment, the authors made some amendments to the existing classification. According to the therapeutical categories identified by the WHO, only patients receiving basic chemotherapy should be followed up in registry group I. Group II is intended for patients with active, including fading, pulmonary tuberculosis who have undergone complete treatment irrespective of its efficiency or who have not received therapy, who require for conditional measures in the foci of tuberculous infection. The developed classification allowed one to reduce the follow-up period and to accelerate clinical recovery and to determine the number of patients in whom rational therapy was ineffective. PMID- 10479939 TI - [New technology of medical service for patients with tuberculosis in outpatient facilities]. AB - The paper summarizes experience in using a new technology of health care provided to patients from antituberculous dispensaries (ATD), which has been developed by the Russian Research Institute of Phthisiopulmonology, Ministry of Health of the Russian Federation. The new procedure was used at the ATD of 114 districts, towns, and cities of 6 Russian administrative regions over 5 years. The areas under study covered more than 7 million adults. The procedure enhances the efficiency of ATD patient service and health care mainly by fixing the optimum time of basic complex therapy of patients with pulmonary tuberculosis (PT), by decreasing the time of recovery by 1-2 years and the follow-up duration for control outpatient groups (III, VII ATD groups), by drastically reducing the number of persons to be medically treated for preventive purposes by 58-70%. The proposed procedure for ATD follow-up care may be useful in the areas of the Russian Federation and requires no material and technical means and resources. PMID- 10479940 TI - [The XXIX International conference on tuberculosis and pulmonary diseases. (IUATLD/UICTMR) Global Congress on lung health. Bangkok, Thailand, November 23 26, 1998]. PMID- 10479941 TI - An annotated bibliography for ethics training in consultation-liaison psychiatry. AB - The expanding field of bioethics has created a need in psychiatry for rapid access to the complex bioethics literature. This is especially true in consultation-liaison work. An annotated bibliography was created by a task force of the Academy of Psychosomatic Medicine charged with exploring how psychiatrists function on bioethics committees. The bibliography is organized into headings that reflect how bioethical problems came to the attention of psychiatrists. Introductory references allow the reader an overview of the history of bioethics and a selection of useful textbooks. References are provided explaining how ethical principles are used. References are also organized by areas of medical work frequently visited by consultation-liaison psychiatrists. PMID- 10479942 TI - Hypochondriasis, somatization, and perceived health and utilization of health care services. AB - The authors determined the different effects of hypochondriasis and somatization on health perceptions, health status, and service utilization in a primary care population. The subjects with hypochondriacal responses (HR) on the Illness Attitudes Scales or high somatic concern (HSC) on the Symptom Questionnaire had a worse perception of health and variably used more health services than the control subjects, even though the HR and HSC subjects had the same level of chronic medical disorders. Regression analyses determined that somatization contributed more to negative health perception and service utilization than did hypochondriasis, although an interaction between the two contributed to the use of psychiatric care. The authors discuss the boundary between hypochondriasis and somatization for its implications for research and clinical practice. PMID- 10479943 TI - Hypochondriasis and somatization related to personality and attitudes toward self. AB - Better definition of the boundary between hypochondriasis and somatization was determined by measuring attitudes to self and personality dimensions associated with these syndromes. In this study, the primary care patients with hypochondriacal responses (HR) on the Illness Attitudes Scales or high somatic concern (HSC) on the Symptom Questionnaire had more negative attitudes to self and more psychological distress than the matched group of primary care control subjects. The HR subjects were different from the non-HR subjects on two of five personality domains on the NEO Personality Inventory (NEO)-Five-Factor Inventory, and the HSC subjects were different from the non-HSC subjects on four of five NEO domains. Analysis of variance demonstrated that somatization explained most of the variance in attitudes, personality, and psychological distress, but hypochondriasis uniquely contributed only to thanatophobia. The authors discuss the boundary between hypochondriasis and somatization and offer a descriptive model of this relationship. PMID- 10479944 TI - Somatic style and symptom reporting in rheumatoid arthritis. AB - The authors studied the relative contributions of psychological characteristics and rheumatoid arthritis (RA) morbidity to RA symptoms and medication side effects. Thirty-one consecutive patients attending an RA clinic completed self report questionnaires and diaries assessing RA symptoms and somatic style, a constellation of beliefs, attitudes, and concerns about disease and health. After 3 months, the patients were assessed for RA symptoms and self-reported medication side effects. At inception, RA symptoms were associated with several components of somatic style. At 3-month follow-up, changes in RA symptoms and the incidence of medication side effects were predicted by somatic style variables measured at inception. The symptoms of RA and the side effects of RA pharmacotherapy are prospectively predicted by somatic style as well as by the severity and extent of RA. PMID- 10479945 TI - Assessment of anxiety disorders in asthmatic children. AB - The study's objective was to determine whether the State Trait Anxiety Inventory for Children, Trait version (STAIC), is suitable for the assessment of DSM-IV anxiety disorders in asthmatic children and adolescents. Ninety-two outpatients were given a semistructured diagnostic interview. They completed STAIC; another questionnaire about anxiety, the Echelle Comportementale d'Anxiete et de Peurs (ECAP); and the Child Depression Inventory. The parents filled in the Child Behavior Check-List (CBCL) and the Conners Parent Rating Scale (CPRS). A group of healthy children was assessed with STAIC. Thirty asthmatic children had anxiety disorders. They had significantly higher STAIC scores than the nonanxious asthmatic and the nonasthmatic children. STAIC scores were independent of age and sex and were correlated with ECAP, CPRS anxiety subscore, CBCL total score, internalizing score, and CBCL anxiety-depression subscore. Internal consistency was 0.75. With a threshold value of 34 for anxiety disorders, this method had a sensitivity of 73% and a specificity of 70%. STAIC was thus a useful method for anxiety disorder screening in a pediatric population. PMID- 10479946 TI - Hematologic side effects of psychotropics. AB - Psychiatrists are often unaware of the potential hematologic complications of the psychotropics they prescribe. Although this review is not a hematologic text reference, relevant hematologic syndromes are described so that the consultation liaison psychiatrist will be familiar with the usual signs, symptoms, and treatments of these syndromes. This article reviews the hematologic side effects of the commonly prescribed psychotropics, including antipsychotics, antidepressants, benzodiazepines, lithium, mood stabilizers (including some of the anticonvulsants), and the acetylcholinesterase inhibitors. Clinical signs and symptoms that should alert the physician to obtain a complete blood count are described. PMID- 10479947 TI - Effects of psychological intervention on panic attack patients in the emergency department. AB - For patients initially seen in the emergency department (ED) for panic attack, this study evaluated the effect of two brief psychological interventions in the ED on later utilization of emergency, psychiatric, and nonpsychiatric medical department services. Each of the two intervention groups received usual ED care, a brochure on panic disorder, and a referral to treatment at the psychiatry department; one of the two groups also received 20-30 minutes of contact with a representative from the psychiatry department. Both intervention groups were compared with a historical control group. The contact condition reduced ED use after the initial visit to the ED, although all three groups had more visits to the psychiatry department and to all nonpsychiatric departments. This decrease was statistically significant (P = 0.0017) when compared with the brochure condition but not when compared with the historical control group (P = 0.0672). The decrease seen in ED use is an important therapeutic and economic finding. PMID- 10479948 TI - Neuropsychiatric toxicity associated with cytokine therapies. AB - The cytokines interleukin-2 and interferon-alpha are potent biological agents used to treat malignancy, infectious diseases, and neurodegenerative disorders. While these medications show substantial therapeutic promise, the neuropsychiatric toxicity associated with these agents is often treatment limiting. The pathophysiology of this toxicity is not well delineated, and adverse effects to the central nervous system are often misdiagnosed by clinicians. This report reviews the preclinical and clinical literature describing the morbidity associated with these agents and suggests appropriate clinical management strategies and future directions for research. PMID- 10479949 TI - Body dysmorphic disorder and a prosthesis. PMID- 10479950 TI - New-onset diabetes mellitus and diabetic ketoacidosis associated with olanzapine treatment. PMID- 10479951 TI - Use of olanzapine in hereditary coproporphyria. PMID- 10479952 TI - Brain injury-induced rapid-cycling affective disorder successfully treated with valproate. PMID- 10479953 TI - Symptoms of patients suffering from somatoform disorders and ifenprodil. PMID- 10479954 TI - [Research on smoking in Spain]. PMID- 10479955 TI - [Methodology used in the original articles on smoking in 4 Spanish medical journals (1985-1996)]. AB - BACKGROUND: Planning being the one factor entailed in doing any research that has the greatest bearing on the quality of the findings thereof, and the cigarette smoking habit being a prime topic for research, a description is provided of the methodological aspects of the original articles on the cigarette smoking habit which have been published in four Spanish medical journals. METHODS: A descriptive study was conducted regarding the original articles on the cigarette smoking habit from Spanish centers which were published in 1985-1996 in the journals Atencion Primaria (Primary Care), Medicina Clinica (Clinical Medicine) (Barcelona), Revista Espanola de Salud Publica (Spanish Public Health Journal) and Revista Clinica Espanola (Spanish Clinical Journal). RESULTS: Most of the 154 original articles studied dealt with the topics of smoking patterns (31.8%) and the cigarette smoking habit as a risk factor (27.3%) and were conducted within a municipal scope (41.7%) or provincial scope (36.1%) with subjects from primary care (71.6%) who visited the doctor (20.8%) falling within the age 25-65 group (30.4%) or over age 15 (27.0%). Surveys were used for collecting the data (38.3%) and were conducted throughout a 12-month period (24.5%) or longer (19.3%). The types most frequently used were descriptive (67.5%), retrospective (79.2%) and cross-sectional (81.8%). CONCLUSIONS: Spanish research on the cigarette smoking habit must cover certain subjects which have not been dealt with in sufficient depth and must invest greater care regarding certain methodological aspects which will improve the quality of the publications, this being a need also brought to fore in other studies. PMID- 10479956 TI - [Extracurricular activities of adolescents useful for smoking prevention programs. OCTOPUS team]. AB - BACKGROUND: The cigarette smoking habit continues to be prevalent to a greater degree than would be desirable among teenagers. Innovative prevention programs are needed. This descriptive cross-sectional study sets out the behavior variables related to the cigarette smoking habit and the extracurricular activities in which teenagers are most frequently involved which are useful for setting out extracurricular prevention programs. METHODS: The data was collected by means of a questionnaire validated in a representative sample of school age youths (ages 10-11 and 13-14) from Asturias. The variables entailed in cigarette smoking were analyzed using the regression method. RESULTS: The starting smoker percentage is 14.5%-42.5%, regular smokers totaling 1.1% and 12.4%, respectively. Two models were constructed with the variables significantly related to smoking behavior, which are properly classified into smoker/non-smoker by 98.85% and 91.39% of the children, by ages. The environmental variables (availability of cigarettes and alcoholic beverages and regular visits to places entailing risk) are the major aspects comprising the model. The most common extracurricular activities are: watching TV, reading and listening to music and watching or playing sports. CONCLUSION: The findings provide keys to planning extracurricular activities tailored to fit in with the activities most popular among teens: TV commercials and ads on music media (CD's, tapes, etc.) and printed information mailed directly to teens at their homes, with messages conveyed by opinion leaders among teens in the fields of sports, music and television. PMID- 10479957 TI - [Prevalence of smoking by women physicians and nurses in the community of Madrid]. AB - BACKGROUND: Tobacco is a major cause of morbidity and mortality in our country. The tobacco smoking by the sanitary staff has a negative influence in the rest of the population. In Spain women smoke more than men. The subject of our study is to identify the tobacco consume in the female sanitary staff in the Community of Madrid. METHODS: Transversal descriptive study in the female sanitary staff in the Community of Madrid, using a conglomerate polystage sampling. RESULTS: We studied 1,235 professionals (435 medicine doctors and 800 nurses). 43.07% of them were smokers, with the higher rate in nurses (47.68%) that are working at hospitals (46.68%). On one hand the tobacco consumption was smaller in medicine doctors between 20-30 years old (22.88%). On the other hand it turned out that nurses in their twenties formed a large group of smokers. A 18.3% of the total sample were actually smokers and a 64.65% had tried to give it up. A 2.93% of woman smoked without taking care if patients was close and 14.94% thought that it had to be allowed to smoke in the hospitals halls. 27.46% of smokers thought that smoking was allowed in their hospitals and 90.84% said that they usually smoked in the sanitary staff rooms, and 30.37% of them answered that specific rooms for smokers were available in their working places. CONCLUSIONS: The prevalence of smoking in the health professionals in the Community of Madrid is very high. Medicine doctors smoke more than nurses, but young nurses smoke than young medicine doctors. Sanitary staff do not obey the no-smoking law in hospitals. The observance of this law is very low. PMID- 10479958 TI - [Evolution of cardiovascular disease mortality in the Toledo Province, 1975 1994]. AB - BACKGROUND: Cardiovascular diseases are the leading cause of death in the developed countries. The cardiovascular mortality varies throughout the different parts of Spain. In this study, an analysis is made of the 1975-1994 trend in the cardiovascular death rate due to heart disease and ischemic heart disease among those falling within the 35-74 age range in the province of Toledo. METHODS: Based on the deaths recorded by the Spanish National Statistics Institute, standardized rates were calculated by the direct method for the causes selected. The trend is assessed based on the mean percentage variance in the rates. The male gender-related ratios are given by causes. RESULTS: A markedly downward trend is found for the cardiovascular death rate as a whole, showing an average drop of 2.58% among males and of 4.78% among females. The decrease is less for heart disease in general, 1.13% among men and 2.68% among women. The trend in the mortality due to ischemic heart disease shows no specific trend, it being possible to term these rates as being stable. For all of the causes, a greater number of deaths among males are recorded, especially with regard to ischemic heart disease, showing a male/female ratio = 1.75. CONCLUSIONS: The cardiovascular mortality in Toledo is dropping similarly to the nationwide rate. The heart disease group as a whole is undergoing a gradual decrease. The stabilization of the death rate due to ischemic heart disease can be explained by the fact that Toledo has low rates as compared to the rest of Spain. PMID- 10479959 TI - [Prevalence of tuberculosis and HIV infections among participants in an intravenous drug user risk-control program]. AB - BACKGROUND: To determine the prevalence of tuberculosis and HIV infection in addition to the related factors among a population of participants in the risk control program in the town of Lleida. METHODS: The sample was comprised of the newly-enrolled participants in the program in April-June 1996, among whom a questionnaire was handed out for collecting the data concerning the variables involved: age, gender, results of the tuberculin test, BCG vaccination, knowledge of the serology regarding HIV, former imprisonment and number of years having used heroin. The prevalence of the tuberculosis and HIV infection was calculated to a 95% confidence interval (CI). The relating of these two variables to all other variables in the study was determined by means of the odds ratio (OR) and its 95% CI. RESULTS: One hundred and fifty (150) patients were seen, 45 of whom were newly enrolled participants. Eighty percent (80%) were males, averaging 31.1 years of age. The prevalence of this dual infection was 8.9% (95% CI 2.8-22.1). The prevalence of the tuberculosis infection was 27.3% (95% CI 12.4-43.0), being higher among former prison inmates (OR = 3.4; 95% CI 0.5-27.4). The prevalence of the HIV infection was 36.1% (95% CI 21.3-53.8), being greater among those who had been using heroin for longer than 11 years (OR = 7.3; 95% CI 1.0-65.9). CONCLUSIONS: Former imprisonment is the main risk factor for tuberculosis infection. The number of years of heroin use are related to the HIV infection, especially when longer than 11 years. The risk control programs in our country should carry out activities aimed at monitoring tuberculosis and HIV infection. PMID- 10479960 TI - [Medical and psychosocial evaluation of a mentally retarded adult population]. AB - BACKGROUND: This study is aimed at describing the psychological, psychopathological, medical and social aspects of a mentally retarded population being provided with care by AFANAS-Jerez affording the possibility of assessing this type of problem from an epidemiological standpoint. METHODS: A descriptive case series study is conducted to characterize a mentally retarded (MR) population. An assessment was made of the psychological, medical and social aspects. The degree of intelligence was analyzed using the Weschler Adult Intelligence Scale. The adaptive behavior, frequency, typology and seriousness of the behavior problems were assessed by means of the Service Planning and Individual Scheduling Inventory. A psychopathological screening examination was conducted. This study is rounded out with a social and medical evaluation by means of structured interviews. The information from the different areas was encoded and was analyzed using the EPIINFO v6.0 and C.I.A. programs. RESULTS: The average age of the mentally retarded population studied was 32.2 (S.D. 9.2 years). Seventy-three percent (73%) males as compared to 27% females. Behavior problems were found to exist among 45.1% of the subjects. We have set up a statistical relationship between the existence/non-existence of said problems and the degree of MR, such that the greater the loss of intellectual capacity, the greater the probability of having behavior problems. One of the psychopathological symptoms worthy of special mention is the anxiety found to exist among 60.2%. The medical examination reveals the large number of problems related to with MR, we thus finding 22.48% of the subjects to have epilepsy, nearly 20% Down's Syndrome, etc. Also worthy of special mention is the low educational level of the parents and the family conflicts resulting from the child's retardation. CONCLUSIONS: Special mention must be made of how MR is a highly complex whole entailing a large number of related medical, psychological, psychiatric and social disorders, this being a situation brought to fore in this study, many of the disorders revealed to exist being related to the degree of MR. We are of the opinion that an epidemiological approach to the study of MR may be of use for a better comprehension of this problem. PMID- 10479961 TI - [Outbreak of shigellosis in a lower-class district]. AB - BACKGROUND: The outbreaks of Shigella sonnei in our environment frequently involve day care centers and elementary schools. An outbreak of shigellosis in a lower-class district is reported. The purpose of this study is that of pinpointing the center of infection, the means of contagion, the characteristics of those infected and of assessing the suitability of the measures taken. METHODS: For monitoring the outbreak over time, a combined observational timeline study was conducted within a territory the bounds of which were marked by means of the conventional epidemiological monitoring variables (time, place and individual). The infectivity of the center of infection (contagion rate) is analyzed by age, gender and school; rate ratio and percentage attributable thereto. RESULTS: On a time-related basis, the outbreak in question started on week 46/97 and ended on week 8/98. This outbreak involved 218 individuals (110 males and 108 females) totaling 5.46% of the district. The highest frequency was found among the 0-4 age group (43.6%), 29.4% in day care (70.32% contagion rate) with a relative risk of 3.9 (95% CI: 2.57-5.93) and 74.36% attributable percentage. The rate ratio between day care and the schools in the district in question is 5.62 (95% CI: 4.33-7.31). Stool cultures were taken and analyzed in 84 cases (38.5%), Shigella sonnei being detected in 38 cases (17.4%). Antibacterial treatment (amoxicillin-clavulan) was set out and individual and group health and safety measures were employed. CONCLUSIONS: The long communicability period and the small number of viable microorganisms necessary for causing this disease fostered its being passed on from one person to another at school and in the home. The measures employed effectively confined the contagion of the infectious agent at the schools. PMID- 10479962 TI - [Tuberculosis in Huesca. Use of the capture-recapture method]. PMID- 10479963 TI - [Making clinical decisions and functional situation in geriatrics. Is there a relationship?]. PMID- 10479964 TI - [Helicobacter pylori and headache]. PMID- 10479966 TI - [Participation in the expert forum "constipation and laxatives" 24 February 1999 in Vienna. Constipation and laxatives]. PMID- 10479965 TI - [Yttrium 90 DOTATOC: a new somatostatin analog for cancer therapy of neuroendocrine tumors]. AB - Current concepts for the treatment of somatostatin receptor positive tumours have not been very motivating up to now. A promising alternative could be the new peptidic vector DOTA-D-Phe1-Tyr3-Octreotide (DOTATOC) recently developed in Basel. It may be labelled with the beta-emitter yttrium-90 (90Y) for internal radiotherapy after systemic application. Pilot therapy studies have shown convincing results with this new radiopharmaceutical. These studies are presented with regard to efficacy and possible toxicity. In summary, the new receptor mediated 90Y-DOTATOC therapy led to tumour response in the majority of patients, and only in some receiving high cumulative doses of > 200 mCi per m2 body surface renal and hematological toxicity due to irradiation occurred. For the reduction of renal accretion, concepts with concomitant amino acid infusions containing L lysine in a higher concentration are currently under way. PMID- 10479967 TI - ["I want to lose weight"]. PMID- 10479968 TI - [Brown vaginal discharge in early pregnancy, hyperemesis. Intact hydatidiform mole]. PMID- 10479969 TI - [Muscle weakness and diffuse muscle pain. Osteomalacia, osteoporosis, L4-L5 intervertebral disc displacement]. PMID- 10479970 TI - [Minor differences in outcome comparing early vs. later initiation of medicamentous blood glucose control in type 2 diabetic patients]. PMID- 10479971 TI - [Symptomatic improvement of patients with nonulcer dyspepsia after Helicobacter pylori eradication]. PMID- 10479972 TI - [Better profile requires professional cooperation]. PMID- 10479973 TI - [Excise the streptococci!]. PMID- 10479974 TI - [Drug utilization among children]. PMID- 10479975 TI - [Utilization of anti-infective agents in pediatric departments in health region 2]. AB - Children are often treated with antiinfective drugs, both in and out of hospital, but few studies of antiinfective drug use in paediatric departments have been published. We have analysed the dispensing of antiinfective drugs from hospital pharmacies to all eight paediatric departments in south-eastern Norway (Health region 2) during the years 1990-95. The total consumption of antiinfective drugs, measured by the number of defined daily doses (DDD), did not increase during the study period, though the total costs for such drugs increased by 48% for all eight departments. In 1995 the antiinfective drug use varied between 15 and 30 defined daily doses per 100 bed days. The total use of cephalosporins increased significantly. For vancomycin, antifungal drugs and antiviral agents, both consumption and cost increased in several departments. Knowledge of the total use of antiinfective drugs may be important when evaluating treatment regimens, especially with regard to microbial resistance. PMID- 10479976 TI - [Drug prescription to 4-year-old children in the county of Vestfold]. AB - We carried out a study of four-year-old children's general health, diseases and use of medicines over a 12-month period in 1995-96. The study was based on a questionnaire administered in connection with the regular health controls. Questionnaires from 1,912 children were returned, representing 79% of children born during one year in the county. More than half the children had received medicine prescribed by a physician, corresponding to 557 preparations per 1,000 children. Some children had used several types of drugs, with a mean of 1.4 preparations per child. Antibiotics were prescribed to 63% of the children. Compared to what has been found in other studies, the use of prescribed medicine was moderate. More pharmacoepidemiological studies are needed to assess whether the use of medicine in children is at an acceptable level. PMID- 10479977 TI - [Antibiotic treatment of acute otitis in children]. AB - Treatment of acute otitis is one of the most common reasons for prescribing antibiotics for children. Recent studies have shown no, or only a small, effect of antibiotic treatment of this condition. We examined the files from the City of Tromso's Emergency Department, which catchment area includes about 12,300 children under the age of 15. During the period from March 1997 to May 1998, 784 children presented with acute otitis, 91.5% of whom received antibiotic treatment. The most frequently drug used drug was penicillin V. Even if there are no or only a small effect of antibiotic treatment of this condition, most children receive such treatment. This study will give baseline data for new studies aimed at reducing the use of antibiotics. PMID- 10479978 TI - [Neurofibromatosis type 1 in childhood]. AB - Neurofibromatosis 1 is one of the most common genetic disorders in man. Although almost every body system can be involved, it most frequently affects the skin, the nervous system and the skeleton. Major disease features are cafe-au-lait spots, axillary/inguinal freckling, neurofibromas and Lisch' nodules. The complications are quite variable, making it impossible to predict the clinical course in an individual patient. To summarize the problems associated with this disease we have quantified manifestations and complications in a group of 38 children with neurofibromatosis 1 who were seen at the University Hospital in Trondheim, Norway between 1987 and 1997. The results are based upon the medical records as well as clinical examination. A wide variety of initial symptoms was recorded, but the majority of the children had cafe-au-lait spots and axillary/inguinal freckling, two of the seven disease features included in the diagnostic criteria given by the National Institutes of Health. The most frequent complications were tumours, with highest incidence of optic pathway gliomas, disturbances of growth and puberty, delayed psychomotor development and learning disabilities. The risk of complications necessitates regular follow-up in a paediatric department. An annual assessment by an ophthalmologist is also recommended, as well as Visual Evoked Potentials/MRI as screening for optic glioma in children under ten years of age. PMID- 10479980 TI - [Possible carcinogenic risk associated with production and use of creosote treated wood]. AB - Creosote is a coal tar product which contains varying amounts of mutagenic and carcinogenic substances such as polycyclic aromatic hydrocarbons (PAHs) and benzene. Marketing and use of creosote and preparations containing creosote, as well as creosote-treated wood, are regulated by a EU Directive. According to the EU classification of such mixtures, inclusion of a warning against creosote as a carcinogen is not necessary if the contents of benzo[a]pyrene (BaP) and benzene are lower than 50 ppm (parts per million) and 1000 ppm, respectively. A recent well designed skin painting study in mice clearly indicates that the creosote preparations had a five-fold higher potency to induce skin cancer than the potency based on BaP content. Furthermore, it was estimated that creosote containing 50 ppm BaP would induce a significant incidence of skin cancer. Preliminary results from determination of concentrations of various carcinogens (BaP and benzene) in the air close to creosote impregnation plants as well as the crudely estimated exposure of children to dermal contact with creosote-treated wood, indicate that the life-time cancer risk from such exposures is in the order of one per 10,000. Despite the uncertainty related to such estimates, these risk levels give reasons for some concern. A further reduction in the content of PAH and benzene in creosote preparations should be considered. PMID- 10479979 TI - [Necrotising fasciitis--a surgical challenge]. AB - Necrotizing fasciitis is a severe, potentially fatal, soft tissue infection. Group A streptococci are the main infectious agents. Early surgical treatment is decisive for the outcome. This report includes 20 patients treated consecutively during a four and a half year period, ten women and ten men, 28 to 78 years of age. Condition at admission to hospital, time from symptom appearance to operation, treatment and results are described. All patients had signs of local infection with rubor and swellings. Severe localized pain was typical. Surgical diagnostic exploration confirmed the diagnosis and led to prompt surgical debridement of all affected tissues. Three patients died, 12 survived without severe sequela and five survived with amputated leg or arm. Necrotizing fasciitis has changed from being a rare disease primarily affecting immunocompromised patients to occurring more frequently also among otherwise healthy individuals. By diseases with local soft tissue infection a liberal use of surgical exploration leads to diagnosis. Increased knowledge of the disease and adherence to a treatment protocol based on antibiotic therapy and mandatory aggressive surgery has probably improved the results with no case fatality in the latter half of the study period. PMID- 10479981 TI - [Do municipalities offer help to children of parents with serious mental illness?]. AB - Children of parents with serious mental illness are considered to be at risk of developing mental illness themselves. Reorganization of the mental health care system has resulted in increased numbers of persons with serious mental illness residing in their home communities, and children have become more directly involved with their parents' illness. A questionnaire was distributed to all of the municipalities in the Mid-Norway health region and to the six city districts in Trondheim in order to determine the number of children of parents with serious mental illness and the health services they routinely receive. 59 (64%) of the 92 questionnaires sent out were returned. 247 children were living with a serious mentally ill parent; six with a mentally ill single father, 102 with a mentally ill single mother. Of the 54 municipalities, six had standard routines for provision of care to the children. Municipalities with such routines attributed this to effective cooperation between professional groups or the special involvement of particular individuals. The study indicates that it is largely a matter of chance whether children do or do not receive help from the communities they live in. PMID- 10479982 TI - [Use of short wave ultraviolet radiation for disinfection in operating rooms]. AB - Over a number of years, short wave ultraviolet radiation (UVC; 200-280 nm) has been used to disinfect air and surfaces in operating rooms, patient rooms, laboratories and so on, as well as air in ventilation ducts. Despite the well documented effect of ultraviolet radiation on air quality, thus reducing the occurrence of infections, this technology has been relatively little used. One advantage of this method is that the UVC sources ensure a continuous reduction in the number of airborne microorganisms that are generated all the time. There are, however, some disadvantages with this method. Human exposure to ultraviolet C may cause keratoconjunctivitis and erythema and requires protection of the skin and the eyes of people exposed to levels above recommended exposure limits. However, by enclosing the UVC sources or by irradiating in the absence of human activity, human exposure is eliminated. These and other aspects concerning the use of short wave ultraviolet radiation as a disinfection agent in operating rooms are discussed in this article. PMID- 10479983 TI - [The patient as a text--metaphors in medicine]. PMID- 10479984 TI - [The patient from an object to a subject]. PMID- 10479985 TI - [Oral glucose load in gestational diabetes--a test that should be used reasonably]. PMID- 10479986 TI - [Increased use of semiautomatic defibrillators in Norway]. PMID- 10479987 TI - [Who is responsible for the occupational environment?]. PMID- 10479988 TI - [Magnesium and leg cramps in pregnancy]. PMID- 10479990 TI - [Back pain]. PMID- 10479989 TI - [Diagnosis of endometrial hyperplasia]. PMID- 10479991 TI - [Pacemaker therapy and driver's licence]. PMID- 10479992 TI - [The unspoken about social matters]. PMID- 10479993 TI - [The psychologists and medicine]. PMID- 10479994 TI - [NSAM's relation towards the Norwegian cohort of newborns/The Norwegian mother and child study]. PMID- 10479995 TI - [Physician or technician?]. PMID- 10479996 TI - The Drosophila glucose transporter gene: cDNA sequence, phylogenetic comparisons, analysis of functional sites and secondary structures. AB - Facilitative glucose transport is mediated by members of the glucose transporter (GLUT) protein family that belong to the large superfamily of twelve transmembrane segment transporters. We have cloned and sequenced a 2,168 base pair cDNA from Drosophila melanogaster (termed Dmglut1: GenBank accession number AF064703) with strong homology to the mammalian Glut genes. The cDNA has an open reading frame encoding a protein of 480 amino acids which shows a similarity of 68% to the human GLUT1 protein. We have done a phylogenetic analysis of the cDNA and the deduced protein sequences and found a significant homology to a putative coding sequence (Ceglut1) in Caenorhabditis elegans. Here we report the results of analyses of functional sites and secondary structures of the proposed proteins and conclude that the Dmglut1 and Ceglut1 genes encode functional glucose transporters. PMID- 10479997 TI - Cytogenetic localization of the growth and reproduction complex (Grc) in the rat and in the mouse and its position in relation to RT1.EC and other loci in the rat MHC. AB - The segment of rat chromosome 20 (RNO20p12) that contains the classical loci of the major histocompatibility complex (MHC; RT1.A-RT1.E) also contains genes affecting growth, reproduction and susceptibility to chemical carcinogens (the Grc) and multiple genes encoding class I MHC antigens (the EC region). The relative positions of the MHC, Grc, and EC region have not been demonstrated explicitly, although they have been postulated from genetic mapping studies. The present study was undertaken to map these regions cytogenetically by several different approaches using cosmids specific for the Rps 18, Hspa1 and Bat1 genes. The order was shown to be: centromere-Rps 18-Hspa1-Bat1-EC-Grc. PMID- 10479998 TI - Environmental effects and the genetics of oviposition site preference for natural yeast substrates in Drosophila buzzatii AB - Habitat selection expressed as oviposition site preferences (OSP), is one component of the complex of behaviours of females seeking a place to oviposit. Drosophila buzzatii females lay their eggs in cactus necroses (rots), where the alternative oviposition sites are patches of adjacent or even partially mixed growing yeast species. The OSP exhibited by individual females is not absolute, but subject to environmental effects and the physiology of the fly, and may vary depending on the particular combination of yeast species present in a rot. Nevertheless, we have shown that OSP of D. buzzatii females is heritable, with evidence from variation among isofemale lines, direct estimation of heritability, generation means analysis and short term selection. Further, this genetic variation appears to be ubiquitous, polygenic and largely non-additive for all yeast species combinations. The consequences of such genotype-specific habitat selection for the maintenance of genetic variation are considered by an evaluation of our results in comparison with assumptions of models of habitat selection. As all assumed mechanisms of these models are apparently met, OSP for yeast species would seem to be a powerful force for the maintenance of genetic variation, and not only at loci affecting the choice of oviposition sites. PMID- 10479999 TI - Detection of Hordeum marinum genome in three polyploid Hordeum species and cytotypes by genomic in situ hybridization PMID- 10480001 TI - Fats: the good, the bad, the trans. PMID- 10480000 TI - Fragile X trinucleotide repeats from a normal population in Rio de Janeiro, Brazil. PMID- 10480002 TI - Making sense of blood pressure medication. PMID- 10480003 TI - Hormone replacement therapy update. PMID- 10480004 TI - Pressure pants relieve chest pain. PMID- 10480005 TI - Fat in food is not the key to feeling full. PMID- 10480006 TI - Alternative medicine after cancer. PMID- 10480007 TI - Designer estrogens: breast cancer benefit, remaining questions. PMID- 10480008 TI - How to be an "upstanding" citizen. PMID- 10480009 TI - Exercise and obesity. PMID- 10480010 TI - Preventing asthma relapses. PMID- 10480011 TI - [Nosocomial pneumonias in a neurology intensive care unit]. AB - BACKGROUND AND OBJECTIVE: Nosocomial pneumonia in patients in an intensive care unit (ICU) are a great problem as a cause of increased morbidity and mortality as well as the resulting high cost of treatment. This study was aimed at determining the incidence of nosocomial pneumonia and the risk factors for its occurrence in patients with severe neurological disease. PATIENTS AND METHODS: Between 1.1. and 31.12.1997, 217 patients (125 men, 92 women; average age 63.4 years) were prospectively included if they were treated for more than 48 hours in the ICU of the Neurology Department of Erlangen University. The occurrence of nosocomial pneumonia (NP) was noted, using the criteria of the Center of Disease Control and Prevention (CDC). Incidence of the diseases was related to age, sex, initial state of consciousness, type of ventilation, duration of stay in the ICU and any associated medical condition. RESULTS: NP was diagnosed in 68 patients (31.4%). Statistically significant relative risks were male sex (2.4 fold, P < 0.01), clouded consciousness with a Glasgow coma score < 8 (6.2 fold, P < 0.001), mechanical ventilation (8.4 fold, P < 0.001), time in ICU > or = 8 days (9.3 fold, P < 0.001) and associated medical condition (3.3 fold, P < 0.005). In 17.7% of cases no relevant pathogen was identified microbiologically. A mixed infection was present in 36.8% of cases. The most common Gram-positive organism was Staph, aureus (35.3%), the most common Gram-negative ones were Ps. aeruginosa (25%), Kl. pneumoniae and Kl. oxytoca (11.8%), E. Coli (10.3%) and Acinetobacter species (7.4%). There was also a high rate of infection or infestation with Candida albicans or glabrata (41.2%). NP played a clinically decisive role in the fatal course of 13 of the 47 patients who died. CONCLUSION: These data (incidence, relative risk) can, by taking into consideration various aspects of specialist and hospital hygienic practices, contribute to a continuing optimization of the prevention and treatment of disease. PMID- 10480012 TI - [Differential diagnosis in anorexia nervosa: glycogenosis II (the Pompe type)]. AB - HISTORY AND ADMISSION FINDINGS: A 29-year-old woman had since early childhood been smaller and thinner than her contemporaries. Her weight was now 36.5 kg, height 158 cm. Anorexia nervosa being suspected she was to be admitted to a psychosomatic clinic. A few days before the date she developed respiratory failure which required mechanical ventilation for a month. Physical examination revealed markedly reduced musculature. INVESTIGATIONS: Muscle biopsy and fibroblast culture indicated acid maltase deficiency (glycogen storage disease Type II [Pompe's disease]). Lung functions were markedly reduced, blood gas analysis revealing global respiratory insufficiency. TREATMENT AND COURSE: Intermittent patient-controlled ventilation (at night by intermittent positive pressure ventilation) clearly improved both her general condition and blood gases. CONCLUSIONS: Acid maltase deficiency should be included as a possible cause of the differential diagnosis of anorexia nervosa. This diagnosis should strictly follow the criteria listed in ICD-10 (International Classification of Diseases), but also after exclusion of other causes. PMID- 10480013 TI - [Dissection of the ramus interventricularis anterior in blunt chest trauma]. AB - HISTORY AND ADMISSION FINDINGS: A 57-year-old man suffered a blunt trauma to his chest when the car he was driving collided head-on with a stationary lorry. He had worn his seat belt and the air-bag had inflated on impact. He was admitted to hospital because of severe left chest pain that started 45 min after the accident. Admission physical examination indicated stable cardiopulmonary status in a fully conscious person with no obvious physical signs, in particular no contusion or other injury to the chest or fractures. INVESTIGATIONS: ECG, echocardiography, chest radiography and routine laboratory tests on admission failed to reveal any diagnostic abnormality. But subsequent ECG and biochemical tests suggested evolution of an acute anterior wall infarction, leading to the patient's transfer to the author's hospital. DIAGNOSIS AND TREATMENT: A coronary angiogram revealed traumatic dissection of the anterior interventricular branch (AIVB) around the origin of the first diagonal branch. There was also suspicion of thrombus formation and clearly impaired left ventricular function with hypo- to akinesia of the anterior wall. Myocardial scintigraphy, positron emission tomography and stress echocardiography were performed to assess the possible need for interventional or surgical revascularization. They demonstrated functioning myocardial tissue in most of the anterior wall so that minimally invasive AIVB bypass operation was planned. CONCLUSION: The possibility of severe injury of the heart and other organs even after primarily blunt chest trauma demands careful follow-up monitoring and, if indicated, extensive noninvasive as well as invasive diagnosis and treatment. PMID- 10480014 TI - [The therapy of Rasmussen encephalitis]. PMID- 10480015 TI - [Colorectal carcinomas in Crohn disease]. PMID- 10480016 TI - [The legal significance of a patient's testament for the physician]. PMID- 10480017 TI - [The Paget-von Schroetter syndrome. On the centenary of the death of Sir James Paget and on the 50th anniversary of the naming of the syndrome]. PMID- 10480018 TI - Changes in colonic mucosal permeability in mouse colitis induced with dextran sulfate sodium. AB - In this study we examined changes in colonic mucosal permeability induced by dextran sulfate sodium (DSS) during the acute phase of mouse colitis. To induce colitis, the mice were given drinking water containing 5% (w/v) DSS (MW = 40,000) ad libitum. Colonic mucosal permeability was evaluated by the permeation of Evans blue (EB) from the lumen into the wall of the colon on 1, 2, 3 and 7 days postadministration of DSS. Mucosal changes were also histologically examined daily for 7 days postadministration. The permeation of EB increased significantly by days 3 and 7 postadministration. Histological analysis showed that crypt loss was the initial change, with no inflammatory process and the surface mucosal epithelial cells remained morphologically intact. These histological changes developed on 2 to 3 days postadministration. Erosion was first recognized at 5 days postadministration. These findings indicated that the increase in colonic mucosal permeability may have occurred in 3 days postadministration, and the increase in mucosal permeability occurred before the appearance of the inflammatory process. This suggests that an increase in colonic mucosal permeability, leading to the destruction of mucosal barrier function, may play an important role in the induction of DSS-induced murine colitis. PMID- 10480019 TI - A parasympathetic ganglion innervating the harderian gland and lacrimal gland of the musk shrew (Suncus murinus): fluorescent tracing and immunohistochemical studies. AB - A small ganglion, named the peri-trigeminal ganglion (PTG), was found in the ventromedial border of the rostral half of the trigeminal ganglion (TG) in the musk shrew (Suncus murinus). In frontal sections, the PTG was semicircular or elliptical in shape. Most of the neurons constituting this ganglion were round in shape and much smaller than those of the TG. The retrograde fluorescent tracer fluoro-gold was injected into various regions of the face in order to investigate innervation by the PTG neurons. When the tracer was injected subcutaneously around the external acoustic meatus and around the circumference of the orbit, a number of labeled neurons were seen not only in the TG but also in the PTG. After applying the tracer to the lacrimal gland (LG) and the harderian gland (HG), numerous labeled neurons were detected only in the PTG. A few labeled neurons were found in the PTG after injection into the palatoglossal arch. Immunohistochemically, most of the neurons constituting the PTG were positive for vasoactive intestinal polypeptide (VIP) antiserum. And a moderate number of somatostatin (SOM)-immunoreactive neurons and a small number of leucine enkephalin (L-ENK)-immunoreactive neurons were detected. Numerous substance P immunoreactive nerve fibers and varicosities were found in the PTG, and fewer L ENK-, SOM- and VIP-immunoreactive fibers were observed. The present results suggest that the PTG is an autonomic ganglion that resembles in part the pterygopalatine ganglion in other species, and mainly innervates the HG and LG. PMID- 10480020 TI - Age-related hematological changes in normal F344 rats: during the neonatal period. AB - In order to clarify age-related changes in hematological values of normal rats after birth, blood samples from neonatal F344 rats of both sexes were examined periodically during the period from 0 to 40 days postpartum. The erythrocyte count (RBC) increased with time after birth as a function of age. In contrast, the reticulocyte count (Retics) continuously decreased with time after birth. Hemoglobin (Hb), hematocrit (Ht), and the mean corpuscular hemoglobin concentration (MCHC) tended to decrease after birth until weaning (about 21 days postpartum), but they began to increase after weaning. Mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) also gradually decreased after birth until weaning, but they were unchanged thereafter. The platelet count (PLT) gradually increased after birth and reached a plateau at weaning. Microscopic examination of blood smears revealed that erythrocytes at birth had characteristic morphological features such as anisocytosis, polychromasia, basophilic stippling, Howell-Jolly body, and erythroblastosis. These characteristic features, however, disappeared by 30 days after birth. The total leukocyte count (WBC) gradually increased with time after birth, due to an increase in the number of lymphocytes. The lymphocyte count started to rapidly increase within several days after birth and the increase continued thereafter. Other differential leukocyte counts also showed various characteristic patterns of changes during the neonatal period. There were no apparent differences between males and females in these changes in hematological values. PMID- 10480021 TI - Renal carbonic anhydrase activity in DBA/2FG-pcy/pcy mice with inherited polycystic kidney disease. AB - DBA/2FG-pcy/pcy (D2-pcy) mice are a hereditary murine model of slowly progressive polycystic kidney disease (PKD) and characterized by the persistent excretion of acidic urine, in association with polyuria, after weaning. In this study, the activity of carbonic anhydrase (CA) and it histological distribution in the kidney of D2-pcy mice were investigated by immunohistochemistry. Significantly higher CA activity was detected in the cytosolic, but not membrane, fraction of kidney homogenates in 5-week-old D2-pcy mice than in age-matched, control DBA/2 (D2) mice, and a more rapid rate of urine acidification was noted in 11-week-old mice when acetazolamide, an inhibitor of the enzyme, was administered orally. By immunohistochemistry for the major renal CA isoenzyme (CA II), epithelial cells in the distal straight tubules and the cortical collecting ducts were stained intensely, whereas those of the proximal convoluted tubules had only weak and diffuse staining. The glomeruli, the proximal straight tubules and the ascending thin limb of Henle's loop were almost free from staining. In the cells lining cysts and/or dilated tubules, CA II activity was well preserved, although the staining intensity was considerably reduced in fully-flattened, lining cells of cysts, but no difference was found between D2-pcy and D2 mice in any segmental localization of renal CA II activity. From these results it seems that D2-pcy mice in the early stages of the cystic disease continue to secrete excess protons through the CA-mediated reaction that is stimulated for regulation of acid-base balance in the distal portion of the nephron and the collecting duct in kidney. It also suggests that monitoring urine pH may be useful in predicting the effects of early interventions on the progression of slowly developing renal cysts. PMID- 10480022 TI - Strain differences of hypertension induced by dietary NG-nitro-L-arginine in normotensive rats. AB - When the potent inhibitor of nitric oxide (NO) synthesis NG-nitro-L-arginine (L NNA) was incorporated into the diet, hypertension was induced and sustained due to the effects of the long-term inhibition of endothelium-dependent relaxing factor (EDRF)/NO. The effects of L-NNA on normotensive rats of four strains (Donryu, Sprague-Dawley (SD), Wistar, and Wistar-Kyoto (WKY)) were compared relative to control rats. L-NNA administration caused a sharp initial increase in systolic blood pressure (SBP) at 2 weeks in all animals, and this was followed by a gradual and steady increase until 4 weeks. At the end of the experiments (5 weeks), the mean SBP of Donryu and SD rats was decreased. The maximum blood pressure of Donryu and Wistar rats during the experiments exceeded 200 mmHg, but that of SD and WKY rats was below 200 mmHg. Body weight loss and death were observed only in L-NNA-fed Donryu rats. Pathological changes in the kidneys and the morbidity rates for the lesions were determined, and indicated that the Donryu L-NNA group was 100% positive. These results suggest that the Donryu strain is more sensitive to L-NNA than the other strains. That dietary L-NNA induced hypertension in normotensive rats of the four strains provides a new artificially-induced hypertensive model in which vasoconstriction occurs mainly due to EDRF deficiency. PMID- 10480023 TI - A new mouse model of spontaneous diabetes derived from ddY strain. AB - By the selective breeding of obese male mice of the ddY strain and using indices of the heavy body weight and appearance of urinary glucose, we established two inbred strains in 1992: one with obesity and urinary glucose (Tsumura, Suzuki, Obese Diabetes: TSOD) and the other without them (Tsumura, Suzuki, Non Obesity: TSNO). The male TSOD mice constantly showed signs of obesity and urinary glucose with increases in food and water intake, body weight and some fat weight. The body mass index (BMI) clearly showed moderate obesity. Increases in the levels of diabetic blood parameters (glucose, insulin and lipids) were also found in males, in which the levels of blood glucose and insulin were high to the ages past the growth peak. In the histological studies, pancreatic islets of the TSOD males were found hypertrophic without any signs of insulitis or fibrous formation. Among these diabetic characteristics, some of which were similar to the reported models of non-insulin-dependent diabetes mellitus (NIDDM), the stable appearances of the hyperglycemia, the hyperinsulinemia and the hypertrophy of pancreatic islets to the ages past the growth peak were the prominent features. In these respect the TSOD mouse may be a useful model for researching the mechanisms of human diabetes and its complications. PMID- 10480024 TI - Delayed development of reflexes and hyperactive locomotion in the spontaneous mutant "waltzing" of the musk shrew, Suncus murinus. AB - The autosomal recessive mutation waltzing (wz), displaying abnormal circling and head-shaking behavior, has previously been reported in the musk shrew (Suncus murinus). Postnatal development of reflexes and locomotor patterns in an open arena were examined in wz/wz mutant shrews. The wz/wz shrews showed extreme developmental delays in surface-righting reflex and negative geotaxis until 10-16 days after birth, but both reflexes eventually recovered to the levels of +/wz normal. Nevertheless, the wz/wz adults exhibited bi-directional circling behavior 59 times, head-tossing behavior 22 times and horizontal head-shaking behavior 6 times more frequent than in the +/wz controls. Although the wz/wz adult shrews were extremely hyperactive with daily spontaneous locomotor activity exceeding 4 7 times control shrew activity, they appeared to have a normal circadian rhythm. This shrew mutant may therefore be useful as a model for hyperactivity syndromes in humans. PMID- 10480025 TI - Glutathione reductase activity and flavin concentration in guinea-pig tissues. AB - Glutathione reductase (GR) activity and flavin concentration were studied in systemic tissues (brain, heart, lung, liver, spleen, stomach, pancreas, muscle, kidney, testis) and blood components (erythrocytes and plasma) from male guinea pigs. GR activity and the flavin concentration were high in kidney and liver, and low in muscle. GR activity in erythrocytes was found in a range of tissues, but flavin concentration in erythrocytes was lower than in any tissues. GR was saturated with flavin adenine dinucleotide (FAD) in almost all tissues, but not in muscle or erythrocytes. PMID- 10480026 TI - Vascular remodeling in hypertensive transgenic mice. AB - We physiologically and histopathologically analyzed vascular damage due to hypertension and vascular remodeling in hypertensive transgenic mice (Tsukuba hypertensive mice; THM). Pubertal (6-week-old) THM already had hypertension similar to blood pressure in adult THM due to an enhanced renin angiotensin system (RAS). They progressively developed remarkable vascular hypertrophy composed of dedifferentiation of vascular smooth muscle cells (VSMCs) and extracellular matrix accumulation in the thoracic aorta, and VSMC hyperplasia was predominant in the abdominal aorta. THM are therefore a useful animal model for studying vascular remodeling mediated by enhanced RAS. PMID- 10480027 TI - Developmental and pharmacological features of mouse emotional piloerection. AB - We demonstrated emotional piloerection in mice given conditioned fear stress by means of a pass-through apparatus. The emotional piloerection was first assessed in mice of different ages. The results showed that the piloerection changed with age. Pharmacological studies showed that the piloerection was inhibited by an alpha 1-adrenoceptor antagonist prazosin, but, surprisingly, was not inhibited by anxiolytic diazepam. These findings strongly suggest that the neuronal system of piloerection is different from that of freezing behavior, and that the neuronal system of piloerection develops with age. PMID- 10480028 TI - Effect of genetic background on establishment of mouse embryonic stem cells. AB - We established 13 embryonic stem (ES) cell lines from 542 embryos crossed between various strains of mice: 10 lines from 129/Sv-ter embryos (10/48, 20.8%) and 3 lines out of other 9 combinations of intra- or inter-strain matings (1 from intracross of C57BL/6CrSlc, 1 from B6D2F1 x C57BL/6CrSlc, 1 from Yok:ddY x Slc:ICR). No ES cell line from 129/Sv-ter x Slc:ICR embryos suggests that ICR strain might have inhibitory genetic factor(s) for the ES cell formation. Some ES cell lines could be obtained from hybrids even if none or few lines from their parental strains, suggesting a heterosis effect can be expected for establishing ES cell lines in mice. PMID- 10480030 TI - [(Symposium of the 78th Hokkaido Medical Congress, 1988) Recent development of information network system in medicine]. PMID- 10480031 TI - [Promotion of information network system for health and medicine]. PMID- 10480029 TI - Linkage mapping of the rat poly(ADP-ribose) glycohydrolase (Parg) gene to chromosome 16. PMID- 10480032 TI - [Experience of telemedicine and information network system at Hokkaido University Hospital]. PMID- 10480033 TI - [Telemedicine and telescience in the field of ophthalmology at Asahikawa Medical College]. AB - Department of Ophthalmology at Asahikawa Medical University has established a video conferencing telemedicine network system between the department and each branch hospital in Japan to transmit full-motion (30 frames/second) color ophthalmological images since October 1994. Nowadays, the department has 15 branches to communicate with our system including one hospital in Fukushima Prefecture; Department of Ophthalmology, Harvard Medical School in USA; Nanjing Medical University in China with the use of INS net 1500 or INS net 64 (3 lines). This system is aimed for telemedicine, telescience as well as supports for ophthalmological surgeries. In this article, we described the present status and future prospective of our telemedicine project for the purpose of the elimination of areas without enough medical facilities and promotion of international scientific collaboration. We also described our "Telemedicine Center" at Asahikawa Medical University which is now under construction. We believe that this technology literally opens up the world to scientific and medical collaboration. PMID- 10480034 TI - [Present telemedicine and its problems in Wakayama Prefecture--from experiences of diagnosing by televised pathology]. AB - The purpose of the telemedicine would be providing advanced medical information for diagnosis and therapy to everyone living in rural districts with proper medical expenses. Telepathology is one of the best solutions for the problem of absolute shortage of number in certified pathological doctors in Wakayama. We employed the computerized microscope (OLMICOSW, Olympus) equipped with remote control system via ISDN, which had a great advantage that pathologists could manipulate the remote microscope and select the appropriate diagnostic field of tissue samples. The telepathology system in Wakayama has produced following results: 1) The system has been well managed by the contract exchanged between the university hospital and a remote hospital. 2) The remote hospital does not need special pathology technician. 3) Official workers in the hospital have deep understanding for the importance of pathological diagnosis through the process on introduction of telepathology system. 4) The telepathology system has improved medical levels in the remote hospital, resulting good contribution to patients and to medical economy of the hospital. PMID- 10480035 TI - [A comparison of alpha 1-acid glycoprotein (AAG) concentration and disopyramide binding in Chinese and Japanese]. AB - The mean concentration of alpha 1-acid glycoprotein (AAG) in plasma and disopyramine binding capacity to AAG (bound disopyramine/AAG) were compared between fourteen Chinese and sixteen Japanese subjects. As a results, the author observed that the mean AAG-concentrations in Chinese and in Japanese were 0.55 +/ 0.11 mg/ml and 0.64 +/- 0.13 mg/ml, respectively. When disopyramine concentration was at 5.0 micrograms/ml, the bound disopyramine/AAG was 7.3 +/- 1.3 micrograms/mg in Chinese and 6.5 +/- 1.3 micrograms/mg in Japanese. Unbound fractions of disopyramine in Chinese and Japanese were 0.23 +/- 0.05% and 0.19 +/ 0.07%, respectively. These results indicated that there was no significant difference in the AAG-concentration and the bound disopyramine/AAG between Chinese and Japanese subjects. In male subjects, however, the AAG-concentration and the bound disopyramine/AAG in Chinese (0.57 +/- 0.14 mg/ml and 7.47 +/- 1.78 micrograms/mg) were significantly different from those in Japanese (0.72 +/- 0.11 mg/ml and 5.84 +/- 0.76 micrograms/mg, p < 0.05), though unbound fraction of disopyramide was not found different between Chinese and Japanese subjects. In female, the AAG-concentration, the bound disopyramine/AAG and unbound fraction of disopyramide did not significantly differ between Chinese and Japanese subjects. These findings suggest a possible structural difference of monosaccharides in plasma-AAG between Chinese and Japanese male subjects. PMID- 10480037 TI - [Establishment of mouse p53 yeast functional assay and evaluation of its detectability of p53 gene mutation]. AB - Mice have widely been used as an experimental model for carcinogenesis induced by chemicals or irradiation. Recently, transgenic mice expressing oncogene proteins and knockout mice lacking for tumor suppressor genes are available, and used for the analysis of mechanisms underlying carcinogenesis. In such experimental carcinogenesis, a rapid and sensitive method for screening p53 mutations is desired. In human carcinogenesis, p53 yeast functional assay has been proved to be a very useful method for screening p53 mutations. However, the p53 yeast functional assay has been unsuccessful in mice because of the high background mutations in them. In the present study, the author developed a mouse p53 yeast functional assay by reducing the background mutations. Initially, 25.8 +/- 2.8% of background mutant red colonies were given by total RNA from normal mouse liver. The background level was lowered to 13.6 +/- 3.3% by improvement of RT-PCR conditions. Then the p53 cDNA-containing plasmids were rescued from the red colonies and the cDNA sequences were determined. The analysis revealed that many background mutations were caused by insertions of extra adenine (A) and thymine (T) at A and T homopolymeric runs, respectively. Majority of the insertion mutations occurred at 5' terminus of murine p53 cDNA. Based on these findings, we constructed a new vector and designed an optimal PCR primer set to exclude 5' terminal sequence in the yeast assay. Finally, the background mutation rate was reduced to 8.0 +/- 1.4%, which was comparable with the rate of 5.2 +/- 2.7% in human p53 yeast functional assay. Using the murine p53 yeast functional assay, we screened several cell lines for p53 mutations and determined those mutations by DNA sequencing. Furthermore, we investigated p53 mutations in UV-irradiated skins of XPC-gene-knockout mice. The yeast functional assay followed by DNA sequencing analysis revealed predominant mutations in dipyrimidines in the p53 coding sequence. These results indicate that mouse p53 yeast functional assay will be very useful for the analysis of p53 mutations in experimental carcinogenesis. PMID- 10480036 TI - [Functional coupling between metabotropic glutamate receptors and G proteins in rat cerebral cortex]. AB - Metabotropic glutamate receptors (mGluRs) are widely distributed in mammalian brain, and are classified into three groups (Group I-III) according to mode of action: by I) homology of amino acid sequence, II) ligand selectivity, and III) signal transduction pathway. The mGluR-mediated signaling cascade is mainly investigated with the use of receptor-expressed non-neuronal cells; and thus, signaling mechanism in the central nervous systems remains to be elucidated. The aim of this study was to elucidate the manner of mGluR-G protein coupling in rat brain by measuring the activities of the high-affinity, low-KM GTPases in G alpha proteins. The selective Group II mGluR agonists, i.e., 2-(2,3 dicarboxycyclopropyl)glycine and (2R,4R)-4-aminopyrrolidine-2,4-dicarboxylate, elevated GTP hydrolysis in a concentration-dependent and saturable manner. On the other hand, the activation of high-affinity GTPase by the selective Group I mGluR agonist, (S)-3-hydroxyphenylglycine, or the selective Group III agonist, L-2 amino-4-phosphonobutylate, was hardly detected. These results indicated that the fashion of mGluR-G protein coupling varies among the mGluR subgroups. It is also suggested that high-affinity GTPase assay is useful for the investigation of mGluR-mediated signal transduction in the brain. PMID- 10480038 TI - [Revisions of the cDNA and primary protein structure of human transcription factor GCF]. AB - GC factor (GCF) was reported as a transcriptional regulator that binds to specific GC-rich sequences in the epidermal growth factor receptor (EGFR) gene promotor, repressing its transcription (Kageyama R. and Pastan I. Cell, 59: 815 825, 1989). In this paper, the author presents revisions of the cDNA and the amino acid sequences of the GCF. 1) 5' rapid amplification of cDNA end (5'RACE) for analysis of RNA of a cancer cell line, A431, was performed, which revealed that the 5' end of GCF cDNA was fused to a 308 bp fragment of other cDNA; simultaneously, the real 5' end cDNA sequence with 31 bp was identified. RNase protection assay presented a main protected band, which was consistent with the result of the RACE analysis. 2) T at the position 787 of the previously reported GCF cDNA was absent from RT-PCR on A431 total RNA. 3) A new sequence with 114 bp was observed on A431 RNA between the positions 851 and 852 of the already reported cDNA by RT-PCR. These observations were confirmed by RT-PCR analyses of RNAs prepared from several other human cell lines, including a non-transformed one (HFL), and white blood cells derived from a normal person. 4) Sequence of genomic GCF DNA was consistent with the new cDNA sequence but not with the previously reported one. 5) The remaining sequence of GCF cDNA was found to be identical to that of the previously reported GCF, based on the results of RT-PCR analyses of RNA prepared from human white blood cells. 6) By the corrections, the GCF cDNA consisted of 2661 bp nucleotides. This revised GCF cDNA (the wild type) encodes a protein of 781 amino acids, including two new sequence regions of 186 amino acids on the N-terminal side of this protein. The revisions eliminated the highly basic region of the amino-terminus of the previously reported GCF, while the other three fourth amino acid sequences of the GCF protein that contained leucine-zipper-like domain had not changed. The revised GCF had no highly homologous protein in the database except the previously reported GCF. 7) The author has developed a specific antibody to human GCF protein. This antibody specifically recognized a protein with a molecular weight of approximately 100 kDa present in the extracts from human cell lines, as confirmed by immunoprecipitation followed by Western blotting. 8) Indirect immunofluorescence of A431 and HeLa cells using the anti-GCF antibody showed that the GCF protein was localized in the nucleus, suggesting that the revised GCF is a nucleoprotein. PMID- 10480039 TI - [Renal, intestinal and whole body metabolic changes in pig LVAD model]. AB - We investigated renal, intestinal, and whole body metabolic changes in response to variations in Left Ventricle Assist Device (LVAD) flow and inspired oxygen concentration (FiO2). Left ventricular assist devices were implanted in 10 adult pigs (weight, 55 +/- 1.76 kg). Renal vein (RV), superior mesenteric vein (SMV), and pulmonary artery (PA) blood oxygen saturation and lactate concentration were measured and used as tissue perfusion markers. These measurements were made before and after changes in LVAD flow or FiO2. Oxygen saturation in PA, SMV, and RV decreased significantly after a reduction in LVAD flow (p = 0.05), with a greater reduction in SMV than in the PA and RV (p < 0.05 at LVAD flow 3.5 l/min; p < 0.01 at LVAD flow 2.0 and 1.0 l/min). The lactate concentration in the PA and SMV increased significantly (p < 0.01) with decreased flow, with a greater increase in the SMV than in the PA (p < 0.05). whereas it remained unchanged in the RV. Oxygen saturation in the PA, SMV, and RV decreased significantly after a reduction in FiO2 (p < 0.05). Lactate concentration in the PA, SMV, and RV increased significantly at FiO2 of 0.10 (p < 0.05). Lactate concentration in the PA and SMV was significantly higher than that in the RV at FiO2 of 0.10 (p < 0.01). The results show that the gastrointestinal tract is at high risk during low perfusion or low FiO2, whereas in the kidneys metabolic function appears to be less disturbed. In clinical practice, this emphasizes the need to ensure adequate blood flow and respiratory function, especially after extubation, in patients with implanted LVAD. This might avoid intestinal ischemia and subsequent endotoxemia. PMID- 10480040 TI - Alternative medicine in radiology. PMID- 10480041 TI - Imagery content during nonpharmacologic analgesia in the procedure suite: where your patients would rather be. AB - RATIONALE AND OBJECTIVES: Imagery as a hypnotic technique can produce analgesia and anxiolysis, but effective use may be restricted to select, highly hypnotizable individuals. This study assessed (a) whether patients not selected for hypnotizability can produce imagery during interventional radiologic procedures and (b) the type of imagery produced. A secondary goal of the study was to familiarize health care providers with a simple, time-efficient technique for imagery. MATERIALS AND METHODS: Fifty-six nonselected patients referred for interventional procedures were guided to a state of self-hypnotic relaxation by a health care provider according to a standardized protocol and script. Patient hypnotizability was assessed according to the Hypnotic Induction Profile test. RESULTS: Patients as a group had average distribution of hypnotizability. The induction script was started in all patients and completed in 53. All patients developed an imagery scenario. Chosen imagery was highly individual, but common trends were nature and travel, family and home, and personal skills. Being with loved ones was an important element of imagery for 14 patients. Thirty-two patients chose passive contemplation, and 24 were action oriented. CONCLUSION: Average patients who present for interventional radiologic procedures and are not preselected for hypnotizability can engage in imagery. Topics chosen are highly individual, thus making prerecorded tapes or provider-directed imagery unlikely to be equally successful. PMID- 10480043 TI - Digital mammography: comparison of adaptive and nonadaptive CAD methods for mass detection. AB - RATIONALE AND OBJECTIVES: The authors compared the performance of adaptive and nonadaptive computer-aided diagnostic (CAD) methods for breast mass detection with digital mammography. MATERIALS AND METHODS: Both adaptive and nonadaptive modular CAD methods employed recent advances in multiresolution and mutiorientation wavelet transforms for improved feature extraction. The nonadaptive method uses fixed parameters for the image preprocessing modules. The adaptive method, a new class of algorithms, adapts to image content by selecting parameters for the image preprocessing modules within a parameter range. Comparison of the two methods was performed for each individual CAD module with a region-of-interest (ROI) database containing all mass types and normal tissue. RESULTS: Receiver operating characteristic (ROC) analysis clearly demonstrated an improvement in performance for the three adaptive modules and a significant overall difference between the two methods. The average ROC area index (Az) values were 0.86 and 0.95 for the nonadaptive and adaptive methods, respectively. The corresponding P value is .0145. For a previously reported database of full mammographic images containing 50 abnormal cases with all mass types and 50 normal images, the adaptive CAD method had a sensitivity of 96% (1.71 false positive results per image) compared with 89% (1.91 false-positive results per image) for the nonadaptive CAD method. CONCLUSION: The adaptive CAD method demonstrated better performance. A study is in progress to determine the generalizability of the adaptive CAD method by applying it to larger retrospective image databases with different film digitizers. PMID- 10480042 TI - Effects of processing conditions on mammographic image quality. AB - RATIONALE AND OBJECTIVES: Any given mammographic film will exhibit changes in sensitometric response and image resolution as processing variables are altered. Developer type, immersion time, and temperature have been shown to affect the contrast of the mammographic image and thus lesion visibility. The authors evaluated the effect of altering processing variables, including film type, developer type, and immersion time, on the visibility of masses, fibrils, and speaks in a standard mammographic phantom. MATERIALS AND METHODS: Images of a phantom obtained with two screen types (Kodak Min-R and Fuji) and five film types (Kodak Min-R M, Min-R E, Min-R H; Fuji UM-MA HC, and DuPont Microvision-C) were processed with five different developer chemicals (Autex SE, DuPont HSD, Kodak RP, Picker 3-7-90, and White Mountain) at four different immersion times (24, 30, 36, and 46 seconds). Processor chemical activity was monitored with sensitometric strips, and developer temperatures were continuously measured. The film images were reviewed by two board-certified radiologists and two physicists with expertise in mammography quality control and were scored based on the visibility of calcifications, masses, and fibrils. RESULTS: Although the differences in the absolute scores were not large, the Kodak Min-R M and Fuji films exhibited the highest scores, and images developed in White Mountain and Autex chemicals exhibited the highest scores. CONCLUSION: For any film, several processing chemicals may be used to produce images of similar quality. Extended processing may no longer be necessary. PMID- 10480044 TI - Simple single-section method for measurement of left and right atrial volumes with electron-beam CT. AB - RATIONALE AND OBJECTIVES: The authors estimated left and right atrial volumes by using a simple method of measurement with nonenhanced electron-beam computed tomography (CT). MATERIALS AND METHODS: One hundred sixty-four contrast material enhanced electron-beam CT studies were divided into two groups. Group 1, which included 104 studies, was used to develop the measurement method (i.e., the formulas) for measuring left and right atrial volumes from a nonenhanced study. Group 2 consisted of 60 studies on which the validity of the method was tested. Measurement of left and right atrial volumes was performed on all section levels by tracing the respective atrial borders for each section, then multiplying the area by section thickness and summing the resultant volumes. RESULTS: Calculated left and right atrial volumes were derived by using the biggest atrial area and cephalic-caudal span. The span was equal to section thickness times the number of sections in which the atria were present. Linear regression analysis formulas were acquired with the biggest atrial area and cephalic-caudal span. Left and right atrial calculated volumes were obtained with these formulas and demonstrated a significant good relation (r > .95, P < .001) and a difference of less than 11% (P < .05) in absolute values between measured and calculated volumes. Intraobserver, interobserver, and interstudy reproducibility were excellent, with less than 10% difference in absolute values. CONCLUSION: Left and right atrial volumes can be accurately estimated from a single midventricular section by using nonenhanced electron-beam CT. PMID- 10480045 TI - Application of pharmacokinetics to electron-beam tomography of the abdomen. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to evaluate the pharmacokinetics of abdominal time-attenuation curves obtained at electron-beam tomography. MATERIALS AND METHODS: Computed tomographic enhancement data of the aorta, portal vein, vena cava, liver, spleen, and pancreas were obtained in 25 patients after injection of 50 mL of contrast medium. These data were used to calculate pharmacokinetic parameters such as half-lives, mean residence times, and areas under the curve with a computer program. RESULTS: Maximal enhancement was observed in the aorta 24 seconds +/- 5 (mean +/- standard deviation) after starting the injection of contrast medium (178 HU +/- 56), in the portal vein after 42 seconds +/- 14 (60 HU +/- 17), in the vena cava after 35 seconds +/- 7 (66 HU +/- 23), in the liver after 58 seconds +/- 15 (24 HU +/- 6), in the spleen after 35 seconds +/- 12 (42 HU +/- 16), and in the pancreas after 39 seconds +/- 15 (42 HU +/- 10). Half-lives of the last phase observed were 108 seconds +/- 123 in the aorta, 33 seconds +/- 30 in the portal vein, 49 seconds +/- 40 in the vena cava, 50 seconds +/- 54 in the liver, 62 seconds +/- 33 in the spleen, and 22 seconds +/- 27 in the pancreas. The computer program allowed for excellent fitting curves to the measured attenuation values and for subsequent calculation of pharmacokinetic parameters. New dosage regimens also could be simulated successfully. CONCLUSION: The pharmacokinetic parameters evaluated might be useful in the optimization of dosing and scanning parameters of the abdomen for ultrafast and helical CT. PMID- 10480046 TI - 1999 Joseph E. Whitley, MD, Award. Facilitating the production of digital radiology teaching files with the radiology annotation and publishing system: a software tool for radiology educators. PMID- 10480047 TI - The current status of intravascular ultrasound imaging. PMID- 10480048 TI - [Susceptibilities of bacteria isolated from patients with lower respiratory infectious diseases to antibiotics (1997)]. AB - The bacteria isolated from the patients with lower respiratory tract infections were collected by institutions located throughout Japan, since 1981. Ikemoto et al. have been investigating susceptibilities of these isolates to various antibacterial agents and antibiotics, and analyzed some characteristics of the patients and isolates from them each year. Results obtained from these investigations are discussed. In these 17 institutions around the entire Japan, 512 strains of presumably etiological bacteria were isolated mainly from the sputa of 440 patients with lower respiratory tract infections during the period from October in 1997 to September in 1998. MICs of various antibacterial agents and antibiotics were determined against 100 strains of Staphylococcus aureus, 81 strains of Streptococcus pneumoniae, 85 strains of Haemophilus influenzae. 71 strains of Pseudomonas aeruginosa (non-mucoid strains), 27 strains of Pseudomonas aeruginosa (mucoid strains), 33 strains of Moraxella subgenus Branhamella catarrhalis, 17 strains of Klebsiella pneumoniae etc., and the susceptibilities of these strains were assessed except for those strains that died during transportation. S. aureus strains for which MICs of oxacillin (MPIPC) were higher than 4 micrograms/ml (methicillin-resistant S. aureus: MRSA) accounted for 55.0%. The frequency of the drug resistant bacteria decreased comparing to the previous year's 67.3%. Arbekacin (ABK) and vancomycin (VCM) showed the most potent activities against MRSA. Imipenem (IPM) and panipenem (PAPM) of carbapenems showed the most potent activities with MIC80S of 0.063 microgram/ml against S. pneumoniae. The frequency of penicillin (PC)-intermediate S. pneumoniae (PISP)+PC resistant S. pneumoniae (PRSP) had decreased gradually, that is, in 1995 the frequency of it was 40.3%, but that was 30.9% in 1997. Against H. influenzae and M.(B.) catarrhalis, all the drugs showed good activities. But the sensitive strains of them against ceftazidime (CAZ) had decreased in 1997, compared those in 1995 and 1996. Meropenem (MEPM), IPM and tobramycin (TOB) showed the most potent activity against P. aeruginosa (mucoid strains). And TOB and ciprofloxacin (CPFX) showed the most potent activities against P. aeruginosa (non-mucoid strains). All drugs except ampicillin (ABPC) were more active against K. pneumoniae in 1997 than that in 1996. Also, we investigated year to year changes in the characteristics of patients, their respiratory infectious diseases, and the etiology. The examination of age distribution indicated that the proportion of patients with ages over 70 years was 45.5% of all the patients showing a slight increase year by year. About the proportion of diagnosed diseases, not so particular changes were recognized as follows: Bacterial pneumonia and chronic bronchitis were the most frequent with 33.6% and 29.1%, respectively. Number of strains isolated from patients before administration of antibiotics were more than those after administration of them in chronic bronchitis, but these had reversed in bacterial pneumonia. The tendency in bacterial pneumonia had been acknowledged since 1995. The increase of S. aureus and P. aeruginosa (both mucoid and non-mucoid strains) isolated after administration of antibiotics, has suggested the decrease of the susceptibility of these strains against antibiotics. Administration of antibiotics has changed the results of the frequency of isolation of bacterial species. Bacterial isolations before administration of antibiotics were as follows: S. pneumoniae 24.5%, H. influenzae 21.4%, S. aureus 18.4% and P. aeruginosa 12.2%. The frequencies of S. aureus decreased after antibiotics administration over 15 days, but the frequencies of P. aeruginosa was not affected. The frequencies of P. aeruginosa was 47.8% after administration over 15 days. From patients administered antibiotics of penicillins and cephems. S. aureus was mainly detected with 31.7-58.3%, and from patients administere PMID- 10480049 TI - [Bacteria isolated from surgical infections and their susceptibilities to antimicrobial agents: special references to bacteria isolated between July 1996 and June 1997]. AB - The annual multicenter studies on isolated bacteria from infections in general surgery and their antimicrobial susceptibility have been conducted in 20 facilities in Japan since July 1982. This paper describes the results obtained during period from July 1996 to June 1997. The number of cases investigated as objectives was 217 for one year. A total of 406 strains were isolated from 177 cases (81.6% of total cases). From primary infections 162 strains were isolated, and from postoperative infections 244 strains were isolated, respectively. From primary infections, anaerobic bacteria were predominant, while from postoperative infections, aerobic Gram-positive bacteria were predominant. Among aerobic Gram positive bacteria, the isolation rate of Enterococcus spp. was the highest. In postoperative infections, the majority of them were Enterococcus faecalis, while in primary infections, many of them were Enterococcus avium. The isolation rate of Staphylococcus spp., especially from postoperative infections, followed that of Enterococcus spp. Among anaerobic Gram-positive bacteria, Peptostreptococcus spp. and Streptococcus spp. were commonly isolated from both types of infections. Among aerobic Gram-negative bacteria, Escherichia coli was the most predominantly isolated from primary infections, followed by Klebsiella pneumoniae and Pseudomonas aeruginosa in this order, and from postoperative infections, P. aeruginosa was the most predominantly isolated, followed by E. coli and Enterobacter cloacae. Among anaerobic Gram-negative bacteria, Bacteroides fragilis group was the majority of isolates from both types of infections. The isolation rate of aerobic Gram-negative bacillus has decreased with time, while those of anaerobes like B. fragilis group and of aerobic Gram-positive bacteria have gradually increased in both types of infections. We found vancomycin resistant strains of neither Staphylacoccus aureus nor Enterococcus spp.; however, the MIC of arbekacin for one of strains of S. aureus was 100 micrograms/ml. Both the MIC90's of meropenem and imipenem/cilastatin against P. aeruginosa isolated in this term were 25 micrograms/ml, which were higher than those against the strains isolated in the previous years. Compared with the isolated strains in the year 1995, progress of resistance against carbapenem antibiotics was confirmed. PMID- 10480050 TI - [In vitro bactericidal activities of new oral penem, faropenem against the various clinical isolates]. AB - The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of faropenem (FRPM) more compared with those of various oral beta-lactams against 15 isolates each of 6 species of microorganism. FRPM possessed potent in vitro antibacterial activity against both aerobic and anaerobic Gram-positive bacteria tested. FRPM showed the same activity as new oral cephems such as cefdinir, cefditoren and cefcapene against all Gram-negative bacteria, but K. pneumoniae strains were less susceptible. The MBC of FRPM against S. pneumoniae and the other strains tested equal and were within 1 dilution the MIC of that, respectively. These results suggest that FRPM has excellent in vitro bactericidal activity against clinical isolates and is a clinically useful for the chemotherapy of bacterial infections. PMID- 10480051 TI - [Bacteriological and clinical break points: evaluation of the usefulness of cefozopran based on MIC]. AB - Drug sensitivity of clinically isolated bacteria to cefozopran (CZOP), which is a new cephem antibiotic, was investigated, and the usefulness of the drug was evaluated from the viewpoint of bacteriological and clinical (pneumonia) break points. The following results were obtained. 1. According to bacteriological break points, methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus spp. showed low sensitivity to cefozopran (CZOP). However, the sensitivity of methicillin-sensitive Staphylococcus aureus (MSSA), E. coli, and Klebsiella spp., which are often isolated as pathogens of common infections, was 100%, that of Enterobacter spp., Serratia sp., and Citrobacter sp. was 90% or higher, and that of Pseudomonas aeruginosa was 80% or higher; the values were comparable to or better than those for ceftazidime (CAZ). These results suggest a expanded antibacterial spectrum and enhanced antibacterial potency of CZOP. 2. The estimated response rate of pneumonia to CZOP was 87.5% in outpatients and 51.9% in inpatients. Therefore, CZOP is considered to be one of the first choices especially in outpatient empiric therapy. PMID- 10480052 TI - [Surgery in the third and fourth age: exploitation or reasonable?]. PMID- 10480053 TI - [Retroperitoneal sarcomas: 2. Surgical and complementary treatment]. AB - Retroperitoneal sarcomas are often voluminous tumors which spread via diverse anatomic paths. The resulting difficult excision explains the high frequency of local or peritoneal recurrence (80% at 10 years). Controlled, complete wide surgical excision is essential as repeated local recurrence is the usual cause of death. The main prognostic factors are the quality and extent of the surgical excision, the histopathology grade, and the metastatic extension. Peroperative pathology is indispensable as no imaging method can provide a sure differentiation between benign and malignant retroperitoneal tumors. CT-scan and MRI are useful to assess extension of the sarcoma to neighboring organs. Surgical treatment should be well-controlled with resectability criteria established preoperatively. A pseudo-capsule limiting the tumor should not be considered as a sufficient safety margin. Extension to neighboring organs is an essential element in preventing the risk of local recurrence. This extension may necessitate associating different access routes. The vascular element must be carefully assessed due to the major problems of reconstruction encountered in subperitoneal tumors, particularly in women. The role of neo-adjuvant chemotherapy has not been well-determined but pre and postoperative chemotherapy protocols currently under investigation have shown objective response. Intraoperative or postoperative adjuvant radiotherapy is also under evaluation. Operated patients should be followed regularly as the risk of recurrence is high. Surgery is indicated for the treatment of recurrences with the best results being obtained after early detection. PET and FDG explorations may be useful for detecting recurrence early. PMID- 10480054 TI - [Ruptures and injuries to the diaphragm]. AB - Injury to the diaphragm is a marker of severe trauma yet remains one of the most misdiagnosed traumatic lesions. The mechanism may be penetrating (stab or gunshot wounds) or blunt (high velocity injuries). Rupture of the diaphragm may be overlooked due to common associated injuries. Diagnosis is based on strong clinical suspicion and repeated chest x-rays. Current diagnostic tools include CT scan and minimally invasive surgical techniques (laparoscopy or thoracoscopy) in stable patients. These relatively new methods could provide more accurate diagnosis. The main feature of diaphragmatic injury is the incidence of late presentation with the possibility of life-threatening complications such as strangulations may even occur late. PMID- 10480055 TI - [Laparoscopic treatment of idiopathic mega-esophagus]. PMID- 10480056 TI - [Treatment of pharyngeal-esophageal diverticula]. PMID- 10480057 TI - [Retroduodenopancreatic detachment]. PMID- 10480058 TI - [Splenic abscess: treatment by percutaneous puncture-drainage]. PMID- 10480059 TI - [Should varicoceles be treated in the adolescent? How?]. AB - Adolescents rarely consult for painful varicocele. The condition has to be confirmed by physical examination and a detailed Doppler exam. An ultrasound may be necessary to measure the size of the testis. About 15% of all adolescents have varicoceles. One out of three is graded II or III in the Dubin and Amelar classification. About 20% of varicoceles graded III occur in association with testicular hypotrophy. We do not know whether boys with a varicocele will fertility problems later on, but only 13% of adult men with varicocele are infertile. Surgery can be considered as necessary only after studying a large number of patients, comparing at random patients operated at a young age and followed for 15-20 years with patients not operated and with a group of healthy controls. The best treatment has to be selected because of the low risk of testicular atrophy and the disappearance of the varicocele in more than 90% of the cases. Inguinal root with microsurgery, and pre or intra-operative radiologic opacifications are the usual choice of most pediatric surgeons. Laparoscopy or retroperitoneoscopy have no major impact on the postoperative results. They are expensive and require great experience. Embolization and other radiological techniques induce a long period of radiation, are not always possible, expensive and demand an experienced radiologist. General anesthesia is required because of the time involved and finally the success rate is low. Microsurgical venous reanastomosis is still confidential probably because of technical difficulties. Scrotal anterograde sclerotherapy is the simplest and cheapest treatment and can be performed with a local anesthetic due to the short time required. But like other procedures, it can induce testicular ischemia. PMID- 10480060 TI - [Emergency admission of a 56 year-old man for occlusive syndrome]. PMID- 10480061 TI - [Management of an abnormal cervical smear. National Accreditation and Health Evaluation Agency]. PMID- 10480062 TI - [National survey on the use of induced labor by obstetricians. Study Group on Induced Labor]. AB - INTRODUCTION: A strong rise in the use of induced labor has been observed in France. The aim of this work was to analyze the different methods used for achieving induction of labor and their implications. METHODS: One out of four French obstetricians were randomly selected to answer a questionnaire on their practice for achieving induction of labor. Four hundred of the 997 obstetricians answered the questionnaire. Univariate and multivariate analysis was applied. RESULTS: A high rate of induced labor was correlated with some areas of the country and with private practice. Certain methods were used in spite of opposing advice by experts in the field: elective induction of labor with unfavorable cervix, use of prostaglandins in elective induction of labor, induction of labor in cases of scarred uterus or breech presentation, use of misoprostol. Some methods were still used in spite of their poor efficacy: intravenous oxytocin used with unfavorable cervix, use of intravensou PGE2. CONCLUSION: This study would show that theory and practice are often distinctly different. Induction of labor is currently used on a far wider scale than ever before. We obviously need studies for careful assessment of the circumstances in which induction of labor is used in order to improve methods and indications of such a clinical practice. PMID- 10480063 TI - [Surgical management of non-palpable breast lesions in ambulatory care]. AB - OBJECTIVE: The aim of our study was to assess if surgery for non-palpable breast lesions could be compatible with a walk-in case hospitalization setting. METHOD: We retrospectively compared 75 patients with a traditional hospital stay to 68 patients with a walking case hospitalization. Overall 143 patients were treated during 1997. Studied parameters were: the quality of the surgical results, the duration of the hospital stay and the post operatives complications. Statistical analysis was realized using the chi 2 test. RESULTS: There was no difference between the studied populations according to the quality of surgical results either the post operatives complications. CONCLUSION: Surgery for non palpable breast lesions can be performed during a one day surgery. The reduction of the duration of the hospital stay decrease the cost of health care system. We should follow the evaluation of the walking case hospitalization for the breast cancer surgery especially when an axillary lymphadenectomy have to be performed. PMID- 10480064 TI - [Evaluation of direct hospital and extra-hospital cost of operative hysteroscopy and vaginal hysterectomy]. AB - The objective of this work was to evaluate hospital and postoperative costs associated with gynecologic surgery for benign uterine pathologies. We studied operative hysteroscopy and vaginal hysterectomy. Hospital costs were broken down into several categories: operating room, postoperative hospitalization, pharmacy surgical instruments, sterilization of reused equipment, food and laundry, indirect costs. The mean total costs for the hospitalization time were 4609.8 francs for hysteroscopic surgery and 7920.3 francs for vaginal hysterectomy. Medical charges in recovery time were calculated with the help of the Social Security. These charges represented 5% of the total cost associated with the procedure. PMID- 10480065 TI - [Prenatal ultrasonic measurements of the eye and the interorbital distance]. AB - OBJECTIVES: The two-fold objective of this study was to ascertain whether the antero-posterior diameter of the fetal eye is comparable to the transversal diameter and to establish nomograms based on the measurements obtained for ocular diameter (OD), mean interorbital distance (MIOD) and the MIOD/biparietal diameter (BPD) ratio related to gestational age. TYPE: A prospective monocentric study based on 398 sonographic fetal eye measurements. RESULTS: The antero-posterior and transverse ocular diameters of the fetal eye remain comparable throughout pregnancy (R = 0.997, p < 0.0001). They were related to gestational age and BPD. The MIOD/BDP ratio decreased with gestational age. The OD/BPD ratio remained nearly constant. Nomograms were established for OD, MIOD and the MIOD/BPD ratio related to gestational age. DISCUSSION AND CONCLUSION: The fetal eye can be measured on any diameter provided the sonographic scan for the measurement is flawless. Nomograms and values are given. Hyper- and hypotelorism and microphthalmia can be found in numerous malformative syndromes. Previously published tables are not well-suited to the French population. PMID- 10480066 TI - [Prediction of fetal hypotrophy by uterine height]. AB - OBJECTIVE: To assess the predictive value of uterine height (UH) in the screening of hypotrophy. MATERIAL AND METHODS: UH percentiles curves depending on gestational age were established on a sample of 385 newborns with normal weight. The assessment of predictive value of the UH was done in 430 pregnant women knowing exactly their last menstruation period. Performance of UH to predict hypotrophy was expressed in terms of sensitivity, specificity, predictive values and likelihood ratios with their 95% confidence intervals. RESULTS: Sensitivity of at least 2 measures of the UH less than the 10th percentile was 71.1% with a specificity of 94.8%. Positive and negative predictive values were 61.5% and 96.6% respectively and the likelihood ratios were 13.6 for a positive test and 0.3 for negative test. CONCLUSION: A serial measure of UH can be useful in the screening of hypotrophy especially in our region where the UH remains the only tool to assess the fetal growth. PMID- 10480067 TI - [Predictive factors of the delivery method in women with cesarean section scars]. AB - OBJECTIVE: To determine the independent effect of clinical and non clinical factors on the mode of delivery after previous cesarean section. METHODS: We performed a retrospective multicenter study of 579 women who had previously undergone a cesarean section and who delivered between January 1995 and June 1997. Maternal and perinatal morbidity associated with trial of labor and elective repeat cesarean was assessed. Multiple logistic regression was used to identify prognostic factors for the outcome of a trial of labor. The odds ratios provided indicate the risk of cesarean section when the factor is present. RESULTS: The rate of successful trial of labor was 74.5%. Overall morbidity was not increased in the trial of labor group. The variables of significant predictive value were the Bishop's score (OR = 15.2 for a score < 3; 95% CI: 5.54 to 41.9), an anomaly of the pelvis (OR = 5.89; 95% CI: 2.37 to 14.7), a previous vaginal delivery (OR = 0.27; 95% CI: 0.12 to 0.60), a fetal distress (OR = 4.11; 95% CI: 2.01 to 8.43), the weight gain during pregnancy (OR = 2.01; 95% CI: 1.10 to 3.68), a delivery between 11 p.m. and 7 a.m. (OR = 0.29; 95% CI: 0.13 to 0.66), a hypertension (OR = 3.10; 95% CI: 1.09 to 8.80) and the use of an intra uterine pressure catheter (OR = 0.26; 95% CI: 0.11 to 0.57). CONCLUSION: A trial of labor should be allowed in most of the women with previous cesarean section. The Bishop's score is the best predictor of the mode of delivery. Induction of labor and a first cesarean for dystocia do not affect the chances of vaginal birth. PMID- 10480068 TI - [Delivery logbook: pertinence of the collected data for scientific exploitation]. AB - OBJECTIVES: To determine if the data recorded in the delivery logbook are relevant and complete compared to the data registered in the patient's medical chart. MATERIAL AND METHODS: Prospective study during one month recording all the data registered by the midwives or the medical doctors in the delivery room immediately after the birth. To compare them to the information collected in the patient's medical chart. Our delivery logbook has 55 headings for each patient. During October 1998, we had 156 births. RESULTS: We had 5.3% of errors divided by missing data and erroneous entries. More precisely, we recorded 9.5% of errors in the antepartum data, 3.2% in intrapartum or postpartum events and 5.8% in the newborn information. CONCLUSION: This study demonstrates that medical data used for scientific projects and recorded in the delivery logbook, have to be interpreted very cautiously. It seems to be reasonable that each obstetrical unit should standardize its registration of data and carry out internal audits. PMID- 10480069 TI - [Fetal and neonatal mortality from 22 weeks of amenorrhea in the Loire area]. AB - BACKGROUND: The Pays de Loire has a low perinatal mortality indicators among French regions but this could be due to under-notification. OBJECTS: To explore this hypothesis we undertook a survey in order to identify all fetal and neonatal deaths occurring at a gestionnal age of 22 weeks or more. We also tried to examine and analyze the causes of death. METHODS: All maternity (26) and neonatal wards (5) in the region took part in the survey in 1995. Clinicians were asked to fill out a questionnaire for all deaths occurring from gestational age (GA) 22 weeks and/or concerning a birthweight of a least 500 g. Only perinatal deaths related to parents living in the Pays de Loire were included in the study. RESULTS: Two hundred and sixty seven perinatal deaths were identified out of a total 29,440 births (9.1 /1000). Eighty three (2.8 /1000) were termination of pregnancy for medical reasons, of which 82% were motivated by chromosomic illness. Ninety-nine stillbirths fell (3.4 /1000) into two GA periods: 24 to 27 weeks (20%) and 38 to 41 weeks (2%). The cause of stillbirths remained unknown in 50% of cases despite a post-mortem examination rate of 87%. There were 29 deaths (1 /1000) in the immediate per and post-partum, 40% of which occurred at GA 22 to 25 weeks. Another 38% occurred at GA 36 to 40 weeks and these were related to undectected malformations or infections. Neonatal and intensive care units reported 56 neonatal deaths (1.9 /1000). GA was under 33 weeks for 44% of them. Deaths were caused by usual complications of severe prematurity, neurologic diseases and malformations. Thirty-two percent of total deaths were not notified to the French Authority: 25% of deaths for termination of pregnancy for medical reasons and 7% for stillbirths and per and post partum deaths. CONCLUSION: This survey suggests that the Pays de Loire perinatal mortality indicators remained low compared with other French regions, even after adjustment for this under notification. This casts doubts on the validity of perinatal mortality monitoring based on official notifications. The cause may lie in the inadequacy of legislation of the particular circumstances of perinatal deaths. PMID- 10480070 TI - [The disinfection of non-autoclavable hysteroscopes in gynecology. Proposition of a protocol and organization of its management]. AB - Iatrogenic infections may result from diagnostic hysteroscopy in case of non autoclavable equipment (such as fibrohysteroscopes) and because of new infectious agents (such as HIV, hepatitis, ...). The equipment should be completely and thoroughly disinfected according to the manufacturer's instructions to prevent infectious complications. PMID- 10480071 TI - [Cutaneous lymphangiectasias acquired after surgical and radiotherapy treatment of breast cancer. Two cases]. AB - We report 2 cases of acquired lymphangiectasias after breast radiosurgical treatment. This well known, rarely reported complication is probably due to a mechanical obstruction of the lymphatic network, and is generally preceded by lymphedema. Our 2 cases, however, did not have previous lymphedema. After a review of the literature, we discuss the role of reparative surgery and other treatment options. PMID- 10480072 TI - [Prenatal and neonatal management of digestive tract duplications. Diagnostic difficulties and therapeutic implications]. AB - OBJECTIVE: To insist on the difficulty of the antenatal diagnosis of digestive duplications, to show the importance of a complete malformative screening, and the need for immediate management at birth. MATERIAL AND METHODS: A cystic duplication of the tongue, a duplication, abdomino-thoracic transdiaphragmatic, and a duplication of the small bowel are described and compared with cases previously reported in th literature. RESULTS: Antenatal evacuation puncture of a duplication of the tongue must be reserved for exceptional situations. Foregut duplications are often associated with other malformations. The duplication of the small intestine can exceptionally expose to a mechanical complication, mostly during the postnatal period, which may require emergency surgery. CONCLUSION: Duplications of the alimentary tract should be detectable on prenatal ultrasound examination. A complete antenatal malformative screening is required and in utero transfer is warranted because emergency surgery may be required. PMID- 10480073 TI - [Anaphylactic shock due to "patent blue" used to search for the sentinel node]. AB - Intraoperative search for the sentinel node using patent blue is considered a non risk procedure. We report the first case of anaphylactic shock and allergy. We emphasize the highly exceptional nature of this adverse effect previously observed in other disciplines using this coloring agent. In light of the favorable outcome in our patient and the importance of the method, this single accident would not be sufficient to contraindicate the continuing use of patent blue in ongoing research. PMID- 10480074 TI - Managing diet quality for Cheddar cheese manufacturing milk. 1. The influence of protein and energy supplements. AB - The effects of supplementing cows' diets with protein and energy on milk composition and the composition and yield of Cheddar cheese were investigated. This research addresses the problems of seasonal reduction in the capacity of cheese curds to expel moisture as observed in parts of south-eastern Australia. Milk was collected from cows offered a basal diet of silage and hay supplemented with different sources and levels of dietary protein and energy. The protein supplements were sunflower, canola, cottonseed meal and lupin, and the energy supplements were maize grain, oats, wheat and barely. This milk was used to manufacture Cheddar cheese on a pilot scale. Cheese moisture content was dependent on the source and level of dietary protein and energy. Milk from cows offered the lupin protein supplements and wheat energy supplements consistently produced cheese with a lower moisture content and moisture in fat-free matter. Milk from these supplemented diets had increased casein concentrations and higher proportions of alpha S2-casein than milk from the poor quality control diet. Cheese yield was directly related to the total casein concentration of milk, but was not influenced by differences in casein composition. Supplementing the cows' diets increased the inorganic P, Mg and Ca concentrations in milk. A low inorganic P concentration in milk from cows offered the control diet was caused by a low intake of dietary P. These findings showed that changes in the mineral and casein composition of milk, associated with diet, could influence the composition of Cheddar cheese. PMID- 10480075 TI - Managing diet quality for cheddar cheese manufacturing milk. 2. Pasture v. grain supplements. AB - The effects of supplementing a basal diet of silage and hay with increasing amounts of harvested spring pasture, or with lupin and wheat, on the composition of milk and the consequent effects on cheese composition and yield were investigated in an indoor feeding study. Milk was collected from five groups of eight cows in mid lactation offered different diets and manufactured into Cheddar cheese on a pilot scale. Milk from cows given the lupin-wheat (LW) and the high pasture level (HP) diets produced low moisture cheese. Cheese produced with milk from cows given the control diet was high in moisture content compared with that made with milk from cows offered the LW diet. Cheese yields from the milk of cows offered the HP and LW diets were greater than from the milk of cows on the control diet, and were associated with the higher casein concentrations of these milks. Casein number was higher in milk from diets supplemented with pasture but was not an indicator of the functional properties of milk that affected cheese moisture. The proportion of beta-casein in milk from cows offered the HP diet was higher and that of gamma-casein lower than in milk from cows given the LW supplement, although cheese moisture content was similar with both diets. Milk from cows offered the HP diet had a greater inorganic P concentration than that from cows given the LW diet, although the dietary intake of P was higher for the LW diet. The significance of the effect of dietary P intake on the concentration of inorganic P in milk and hence its suitability for cheesemaking was apparent when dietary P intake was low, as shown in milk produced by cows offered the control diet. PMID- 10480076 TI - Usefully combining a series of unreplicated cheesemaking experiments. AB - Applied dairy research is characterized by experiments for which financial and physical constraints permit only a small number of experimental units. With few units it is difficult to replicate treatments, and without replication experimental error cannot be estimated. The statistical analysis and interpretation of such experiments is problematic. However, if there have been several such experiments it may be possible to perform a combined analysis. Nine unreplicated experiments comparing effects of diet on the composition of cows' milk and on cheese characteristics were jointly analysed as an incomplete block design. This analysis method was contrasted with analyses of individual experiments. For cheese moisture, the key outcome measurement, the assessment of statistical significance concurred for the two methods in 13 out of 21 comparisons of treatments with the control. Sources of error variation allowed for under the two methods were delineated. The combined analysis paradigm provided stronger inference and a wider interpretation of results than could be achieved using analyses for individual experiments. Unequal replication of treatments and unequal concurrence of treatments within experiments over the series gave rise to a wide range of SED. The challenge of presenting results with unequal SED was addressed graphically using error bars. Attention to series design, in particular the apportioning of replication and treatment concurrence across the series of experiments, was shown to ameliorate presentation difficulties and, more importantly, to yield higher precision at no extra cost. PMID- 10480077 TI - Respiratory burst activity in activated and unstimulated isolated bovine blood neutrophils during experimentally induced Escherichia coli mastitis. AB - The respiratory burst activity, measured as H2O2 production, of isolated bovine polymorphonuclear leucocytes (PMN) was evaluated during experimentally induced Escherichia coli mastitis by means of flow cytometry in cells activated by phorbol 12-myristate 13-acetate (PMA) and in unstimulated cells. As expected, a significantly reduced respiratory burst activity was observed in PMA-activated PMN 18 h after intramammary inoculation with Escherichia coli. At this time only 75% of the PMA-activated PMN showed a respiratory burst, but with a higher intensity than that measured before and later after infection with Esch. coli. In addition, an increase in the respiratory burst activity was observed in unstimulated blood PMN during a short period at 18 h after infection, when up to 30% of the unstimulated PMN had a respiratory burst activity. The increase in the respiratory burst intensity of PMA-activated PMN and the spontaneously augmented production of reduced oxygen species by the unstimulated PMN during infection with Esch. coli might indicate the production of a natural stimulator of burst activity in circulation, most probably originating from the inflamed udder. PMID- 10480078 TI - Characteristics of transport systems of L-alanine in mouse mammary gland and their regulation by lactogenic hormones: evidence for two broad spectrum systems. AB - The characteristics of the transport systems of L-alanine in lactating mouse mammary gland and their regulation by lactogenic hormones have been studied. L alanine uptake was mediated by three Na(+)-dependent and one Na(+)-independent systems. The 2-(methylamino)isobutyric acid-sensitive component of Na(+) dependent uptake exhibited the usual characteristics of system A. Cl- dependency has been established for system A. The other two Na(+)-dependent systems, which we have named BCl(-)-dependent and BCl(-)-independent, are described for the first time. These are systems with broad specificity and were distinguished on the basis of inhibition analysis, Cl- dependency and the effect of preloading mammary tissue with amino acids. The Na(+)-independent route was identified as system L, which operates independent of Cl-. The A, L and BCl(-)-independent transport systems were upregulated in pregnant mouse mammary tissue cultured in vitro in the presence of lactogenic hormones (insulin plus cortisol plus prolactin). Insulin alone also upregulated systems A and L to some extent in pregnant mouse mammary tissue. BCl(-)-dependent activity was not detected in pregnant mouse mammary tissue and was not induced by lactogenic hormones in vitro. PMID- 10480079 TI - Effects of sheep alpha s1-casein CC, CD and DD genotypes on milk composition and cheesemaking properties. AB - The effects of sheep alpha s1-casein CC, CD and DD genotypes on milk composition and cheese yield were studied. Processed bulk milk was collected from three groups of 15 ewes, carrying alpha s1-casein CC, CD and DD genotypes. CC milk was higher in casein content than CD or DD milk (+3.5 and +8.6% respectively), and had a higher protein: fat ratio and a smaller casein micelle diameter. In addition, DD milk had a significantly lower alpha s1-casein content. The main differences were in curd formation: CC milk had better renneting properties. Cheesemaking trials, carried out in a pilot plant, showed that CC milk had better cheesemaking characteristics than DD milk, while CD milk was intermediate. Both 1 d old and fully ripened cheeses had different fat: dry matter ratios and alpha s1 I-casein electrophoretic mobilities: these were lower for DD cheese. As a consequence, these genotypes could be considered as markers of milk and/or cheese quality. PMID- 10480080 TI - Bovine immunoglobulin G, beta-lactoglobulin, alpha-lactalbumin and serum albumin in colostrum and milk during the early post partum period. AB - Colostrum and milk samples from 60 Holstein-Friesian cows were analysed for concentrations and yields of immunoglobulin G (IgG), beta-lactoglobulin (beta lg), alpha-lactalbumin (alpha-la) and serum albumin (BSA) throughout the first 16 milkings post partum (8 d of lactation) using a single radial immunodiffusion assay. Concentrations (mg/ml, means +/- SD) at first milking were IgG 59.8 +/- 28.5, beta-lg 14.3 +/- 4.6, alpha-la 2.04 +/- 0.6, BSA 1.21 +/- 0.44. Large variations were recorded for IgG concentrations (15.3-176.2 mg/ml) and yields (0.2-925 g). Cows in their first lactation produced significantly lower concentrations and yields of colostral IgG than cows in later lactations. A colostral yield of IgG below the 100 g required to prevent calf hypo-gamma globulinaemia was found in 18.3% of the cows. The concentrations of IgG, beta-lg and BSA dropped abruptly in subsequent milkings and alpha-la concentration decreased slowly. The mean IgG concentration was < 2 mg/ml after eight milkings and < 1 mg/ml after fifteen milkings. However, IgG concentration did not differ significantly, at the 1% level, during milkings 11-15. The results were tabulated to make it possible to calculate the excess of whey proteins that would be obtained if early milks were illegally added to the milk supply. PMID- 10480081 TI - Novel angiotensin-I-converting enzyme inhibitory peptides derived from recombinant human alpha s1-casein expressed in Escherichia coli. AB - Recombinant human alpha s1-casein expressed in Escherichia coli was purified and digested with trypsin in an attempt to find peptides with angiotensin-I converting enzyme (ACE) inhibitory activity. Three novel ACE inhibitory peptides, A-II, B-II and C, were isolated and their amino acid sequences identified as Tyr Pro-Glu-Arg (residues 8-11), Tyr-Tyr-Pro-Gln-Ile-Met-Gln-Tyr (residues 136-143) and Asn-Asn-Val-Met-Leu-Gln-Trp (residues 164-170) respectively. ACE inhibitory activities were measured for the corresponding synthetic peptides, and the ACE IC50 (the amount of peptide causing 50% inhibition of ACE activity) values of A II, B-II and C estimated to be 132.5, 24.8 and 41.0 mumol/l respectively. Peptides A-II and C were resistant to further digestion by pepsin, whereas peptide B-II was hydrolysed. All three peptides were resistant to digestion by chymotrypsin. These ACE inhibitory peptides may prove useful for oral administration in the treatment of hypertension. PMID- 10480082 TI - Chemiluminescent determination of xanthine oxidase activity in milk. AB - A chemiluminescent method for determining xanthine oxidase (XOD) activity was developed and applied to the assay of milk enzyme activity using a photomultiplier luminometer. Various kinds of milk and cream samples were analysed for XOD content. In pasteurized milk, XOD activity depended on the fat content and in UHT milk it disappeared owing to the heat treatment. Milk sample preparation was very simple, requiring only homogenization at 40 degrees C followed by a 1:10 dilution with UHT ('XOD-free') milk. The assay was carried out at 25 degrees C. The response obtained from XOD standard solutions in milk was linear from 0.1 to 500 enzyme units (U) l-1, but for the actual milk samples values ranged only from 1 to 135 U l-1. The detection limit at 2 SD was 0.1 U l-1 in milk, while in buffer it was 100 times lower. The intra-assay and interassay CV for XOD activity in milk were 6-12%. PMID- 10480083 TI - Protein A gene polymorphism analysis in Staphylococcus aureus strains isolated from bovine subclinical mastitis. PMID- 10480084 TI - Estimating the costs associated with endemic diseases of dairy cattle. PMID- 10480085 TI - Free radical formation in freeze-dried raw milk in relation to its alpha tocopherol level. PMID- 10480086 TI - ADSA Foundation Scholar Award. Critical issues affecting the future of dairy industry: individual contributions in the scope of a global approach. AB - Several constraints that have been affecting the dairy industry are identified in a critical fashion, and directions are given with an emphasis on food processing implemented at the postproduction level. The rationale for modifications aimed at enhancing the appeal of condensed dairy products should be consubstantiated in strengthening of organoleptic characteristics, improvement of nutraceutical impact, and reduction of polluting power. This enumeration follows an order of increasing time scale required for consumer perception and increasing size scale associated with expected impact. Pursuance of such streamlines should lead to manufacture of dairy products that resemble nature more closely in terms of milk coagulation, milk fat modification, milk fermentation, whey fermentation, and starter culture addition. Directions for research and development anticipated as useful and effective in this endeavor, and which have been previously and consistently adopted in the development of an individual research program, are characterization and development of alternative rennets from plant sources, development of starter and nonstarter cultures from adventitious microflora, utilization of probiotic strains as starter cultures, upgrading of whey via physical or fermentation routes, and modification of milk fat via lipase-mediated interesterification reactions. PMID- 10480087 TI - Purification and identification of potentially bioactive peptides from enzyme modified cheese. AB - Antihypertensive peptides inhibiting angiotensin I-converting enzyme have been isolated from enzymatic hydrolysates of various food materials, but no information is available on the isolation of antihypertensive peptides from enzyme-modified cheese. In this study, several bioactive peptides, mainly potential antihypertensive peptides from enzyme-modified cheese prepared by commercial and Lactobacillus casei enzymes, were purified and identified. Enzyme modified cheese samples were prepared by combination of Neutrase (1883.0 U/ml), L. casei enzymes (amino peptidase activity 86.4 leucine aminopeptidase U/g), and Debitrase (22.0 leucine aminopeptidase U/g). The water-soluble fractions of the enzyme-modified cheeses that were prepared by different enzymes were subjected to reverse-phase HPLC on a Delta Pak C18 column. Each peak was purified on the same column using a binary gradient. One peak from the Neutrase digest, five peaks from the Neutrase-Debitrase digest, and two peaks from the Neutrase-Lactobacillus enzyme digest were purified and identified by API mass spectrometry. On the basis of their molecular masses, amino acid sequences of purified peptides were identified. beta-Casomorphin with a sequence like that of beta-casein (YPFPGPI f 60-66) was found after the Neutrase digest. All of the peptides purified from the digests with combination of Neutrase and Debitrase or Neutrase and L. casei enzymes contained active sites in their sequences. The presence of sites containing potential antihypertensive peptides suggests that the purified peptides may have antihypertensive properties. Thus, the enzyme-modified cheese process, mainly designed to produce flavor ingredients, may simultaneously produce bioactive peptides, which are considered to be of physiological importance. PMID- 10480088 TI - Reactive oxygen species and lipid peroxidation product-scavenging ability of yogurt organisms. AB - The antioxidative activity of the intracellular extracts of yogurt organisms was investigated. All 11 strains tested, including five strains of Streptococcus thermophilus and six strains of Lactobacillus delbrueckii ssp. bulgaricus, demonstrated an antioxidative effect on the inhibition of linoleic acid peroxidation. The antioxidative effect of intracellular extracts of 10(8) cells of yogurt organisms was equivalent to 25 to 96 ppm butylated hydroxytoluene, which indicated that all strains demonstrated excellent antioxidative activity. The scavenging of reactive oxygen species, hydroxyl radical, and hydrogen peroxide was studied for intracellular extracts of yogurt organisms. All strains showed reactive oxygen species-scavenging ability. Lactobacillus delbrueckii ssp. bulgaricus Lb demonstrated the highest hydroxyl radical-scavenging ability at 234 microM. Streptococcus thermophilus MC and 821 and L. delbrueckii ssp. bulgaricus 448 and 449 scavenged the most hydrogen peroxide at approximately 50 microM. The scavenging ability of lipid peroxidation products, t-butylhydroperoxide and malondialdehyde, was also evaluated. Results showed that the extracts were not able to scavenge the t-butylhydroperoxide. Nevertheless, malondialdehyde was scavenged well by most strains. PMID- 10480089 TI - Management practices associated with the incidence rate of clinical mastitis. AB - Risk factors for the incidence rate of clinical mastitis were studied in 274 Dutch dairy herds. Variables that were associated with resistance to disease were the feeding, housing, and milking machine factors. Variables that were associated with exposure were grazing, combined housing of dry cows and heifers, and calving area hygiene. Postmilking teat disinfection in herds with a low bulk milk somatic cell count and years of practicing dry cow therapy were positively associated with the incidence rate of clinical mastitis. Herds with a low bulk milk somatic cell count and in which postmilking teat disinfection was not used had lower incidence rates of clinical mastitis than did other herds. The incidence rate of clinical mastitis caused by Escherichia coli was mostly related to housing conditions, hygiene, and machine milking. The incidence rate of clinical mastitis caused by Staphylococcus aureus was mostly related to factors associated with bulk milk somatic cell count and factors that might be due to cause and effect reversal. A strong positive correlation existed between the incidence rate of clinical mastitis caused by Streptococcus dysgalactiae and the incidence rate of clinical mastitis caused by Staph. aureus. The incidence rate of clinical mastitis caused by Streptococcus dysgalactiae was related to nutrition, milking technique, and machine milking. The incidence rate of clinical mastitis caused by Streptococcus uberis was associated with factors related to housing, nutrition, and machine milking. PMID- 10480090 TI - Management style and its association with bulk milk somatic cell count and incidence rate of clinical mastitis. AB - Management style and its association with bulk milk somatic cell count (SCC) and the incidence rate of clinical mastitis were studied in 300 Dutch dairy herds. Cluster analysis was used to identify groups of farmers who had similar management styles for the prevention of mastitis. Two groups of farmers could be differentiated. The management style of the first group of farmers was described as clean and accurate; the management style of the second group of farmers was described as quick and dirty. The relationship between clusters and the bulk milk SCC category was high. The relationship between clusters and incidence rate of clinical mastitis was weak. Compared with herds with a high (250,000 to 400,000 cells/ml) bulk milk SCC, herds with a low bulk milk SCC (< or = 150,000 cells/ml) were managed by farmers who were younger, had children with a higher education, and were more eager to invest. Farmers of herds with a low bulk milk SCC kept better records and were more familiar with each cow in their herds. The most striking difference between farmers of herds with low and high bulk milk SCC was that the first group worked precisely rather than fast; the latter group of farmers worked quickly rather than precisely. As a result, the farms with herds that had a low bulk milk SCC had better hygienic conditions than those farms with herds that had a high bulk milk SCC. We also discuss the implications for producer education with regard to udder health. PMID- 10480091 TI - Comparison of seven antibiotic treatments with no treatment for bacteriological efficacy against bovine mastitis pathogens. AB - Milk culture results were retrospectively reviewed from 9007 cases of subclinical mastitis affecting cows housed in dairy herds located in New York and northern Pennsylvania. Cases included in this analysis had at least one mastitis pathogen isolated from the initial milk sample, were recultured within 1 mo, had permanent cow identification, and had records of whether mastitis was treated with an antibiotic or no treatment at all. Overall bacteriological cure rate for 21 mastitis pathogens was 68% (6097 of 9007). Antibiotic treated cases had a higher cure rate (75%) than did untreated cases (65%). Antibiotic treatments that significantly differed from the untreated cure rate of 65% were amoxicillin (82%), erythromycin (76%), cloxacillin (73%), and pirlimycin (44%). Cure rates for antibiotic treatments with cephapirin, hetacillin, or penicillin did not differ from the untreated cure rate. Agents for which some antibiotics were associated with increased cure rates compared with no treatment were Streptococcus agalactiae, streptococci other than Strep. agalactiae, and coagulase-negative staphylococci. The antibiotic most commonly associated with higher cure rates was amoxicillin. Most of the 21 mastitis agents showed no difference in bacteriologic cure rates between any of the 7 antibiotic treatments and no treatment. PMID- 10480092 TI - Effect of recombinant bovine somatotropin on milk production and composition of cows with Streptococcus uberis mastitis. AB - The protective effect of bovine somatotropin (bST) during experimental Streptococcus uberis mastitis in cows was studied. The left quarters of 10 cows were infected with 500 cfu of S. uberis O140J. Five cows were subcutaneously treated with 500 mg of recombinant bST 7 d before and after infection, and 5 control cows received the excipient. In the treated cows, total milk production significantly increased after the first and second bST treatments. After infection, milk production decreased 24 and 40% in the infected quarters, 6 and 14% in the uninfected quarters, and 15 and 28% overall for treated and control cows, respectively. In the bST group, milk production was completely restored after 3 wk, but, in the control group, total production and the production of the infected quarters remained lower than preinfection production. The increase in somatic cell count occurred earlier and more rapidly in the control group, and the return to normal values was also more rapid in these cows. The amount of bacteria in milk was higher in the control cows. Changes in milk composition, such as lactose, protein, fat, Na+, K+, and Cl-, were significantly more pronounced in the control cows. Also, clinical symptoms were more prominent in the control cows. Somatotropin protected the mammary gland from excessive production losses and compositional changes during a subsequent episode of experimentally induced Streptococcus uberis mastitis and significantly improved the normalization of production and composition, which indicates a beneficial effect on the restoration of the integrity of the blood-milk barrier. PMID- 10480093 TI - Clinical mastitis associated with abortion in dairy cows. AB - The objective of this study was to assess the association between clinical mastitis and abortion during early gestation in lactating dairy cows. During the study period, there were 2087 cows diagnosed pregnant, 60 cases of clinical mastitis, and 127 cases of abortion. Logistic regression was used to evaluate the association between the occurrence of mastitis and subsequent abortion. This study found that cows that had clinical mastitis during the first 45 d of gestation were at 2.7 (95% confidence interval = 1.3 to 5.6) times higher risk of abortion within the next 90 d than were cows without mastitis. Cows with a calving to conception interval > 174 d were at lower risk (43%) of abortion compared with cows with a calving to conception interval < 79 d (odds ratio = 0.6; 95% confidence interval = 0.3 to 0.9). Conception on a breeding detected by secondary signs of estrus was also significantly associated with increased risk of abortion (odds ratio = 1.7; 95% confidence interval = 1.1 to 2.8). No significant association was found between parity, breeding season, or milk production or use of prostaglandin F2 to induce successful estrus and subsequent abortion. PMID- 10480094 TI - Bacterial counts associated with sawdust and recycled manure bedding treated with commercial conditioners. AB - Bacteria counts associated with untreated organic bedding materials were compared with those of bedding treated with either an alkaline commercial bedding conditioner, acidic commercial bedding conditioner, or hydrated lime. Bedding materials were recycled manure and kiln-dried sawdust. The effects of bedding treatments on bacteria counts differed between bedding types. Each of the bedding treatments significantly reduced bacteria in recycled manure prior to use. The alkaline conditioner and hydrated lime effectively inhibited bacteria in recycled manure for 1 d. Bedding counts and teat swabs of cows housed on recycled manure treated with the alkaline conditioner were reduced on d 2. The use of the acid conditioner in recycled manure had little effect on bacteria in bedding. Sawdust differed from recycled manure in that bacteria in untreated sawdust prior to use were minimal, and populations increased rapidly during the first 2 d after use as bedding. The acid conditioner had a bacteriostatic effect in sawdust, evident by the reduction of bacteria on d 2. The alkaline conditioner and hydrated lime did not alter bacteria counts in sawdust compared with untreated sawdust. Antibacterial activity of each conditioner deteriorated between d 2 and d 6 in both beddings. The antibacterial activities of conditioners were related to the pH of bedding materials. The use of commercial bedding conditioners initially reduced bacterial counts; however, the antibacterial effects had diminished between d 2 and 6 after use in bedding. PMID- 10480095 TI - Shifts in bovine CD4+ subpopulations increase T-helper-2 compared with T-helper-1 effector cells during the postpartum period. AB - This study determined the cytokine profile of CD4+ T-helper cells to elucidate the specific CD4+ T-helper phenotype during the postpartum period. Peripheral blood mononuclear cells were isolated from cows during periods of increased susceptibility (3 d postpartum, n = 7) and decreased susceptibility (mid- to late lactation, n = 6) to mastitis. Isolated mononuclear cells were magnetically separated into CD4(+)-enriched or CD4(+)-depleted populations using specific bovine monoclonal antibodies and were confirmed to be enriched or depleted by flow cytometric analysis. T-helper-1 and T-helper-2 subpopulations were distinguished by cytokine profiles, at both the molecular and protein level, by competitive quantitative reverse transcriptase-polymerase chain reaction and specific bioassays, respectively. The CD4(+)-enriched cultures isolated postpartum had enhanced interleukin-4 and interleukin-10 mRNA transcript expression; cultures isolated during the mid- to late lactating period had enhanced interleukin-2 mRNA transcripts. Depletion of CD4+ lymphocytes decreased, and enrichment of CD4+ lymphocytes increased interferon-gamma transcripts in cultures isolated from mid- to late lactation cows. Interferon-gamma and interleukin-2 bioassays revealed that cytokine secretion paralleled mRNA transcript levels. These data suggest that CD4+ lymphocytes act predominantly as T-helper-2 compared with T-helper-1 within 3 d after calving. Alterations in the T-helper-1 and T-helper-2 responses, and therefore the repertoire of cytokines produced, may be an underlying reason for diminished host immune response during the postpartum period. PMID- 10480096 TI - Anti-idiotypic responses of lactating cows immunized with monoclonal antibodies against bovine somatotropin. AB - The objective of this study was to produce anti-idiotypic antibodies with bovine somatotropin (bST)-like activity by active immunization of lactating cows and to determine their effects on milk yield. Several monoclonal antibodies against bST were evaluated for their interaction with bST in a rat growth bioassay. Two bST agonist monoclonal antibodies (1 and 2), and two bST-antagonist monoclonal antibodies (3 and 4) were selected. Cows were immunized with immunoglobulin G as a control (n = 12) or with one of the four anti-bST monoclonal antibodies (1, 2, 3, 4; n = 12) on d 3, 24, 45, 66, 87, 108, 129, and 150 of lactation. From wk 3 of lactation, all cows immunized with each of the four anti-bST monoclonal antibodies developed anti-idiotypes until wk 30 of lactation. Total lactation yields were not different among monoclonal antibodies 2, 3, and 4 and control cows (9299, 9321, 9733, and 9415 kg, respectively). However, cows immunized with anti-bST monoclonal antibody 1 had reduced lactation yield compared with cows on other treatments (8136 kg). Daily milk yield of cows immunized with monoclonal antibody 1 was decreased from wk 9 of lactation [36.2 vs. 40.9 kg/d (control)] until the end of lactation, concomitantly with decreased bST concentration from wk 9 of lactation. Cows immunized with anti-bST monoclonal antibody 4 had increased milk yield compared with that of controls during wk 3 to 6 and wk 18 to 21 of lactation. Therefore, anti-idiotypes directed against anti-bST 1 had bST antagonistic effects on lactation performance; anti-idiotypes against anti-bST 4 transiently increased milk yield. PMID- 10480097 TI - Effects of long daily photoperiod and bovine somatotropin (Trobest) on milk yield in cows. AB - Bovine somatotropin (bST) and exposure to long daily photoperiod increase milk yield of dairy cattle. We tested the hypothesis that long daily photoperiod and bST treatment would increase milk yield in an additive manner in lactating cows. At winter solstice, 40 lactating cows were started on a 140-d experiment; cows were greater than 70 d in milk (DIM) and were balanced for uniformity of DIM and milk yield within parity. Cows were randomly assigned to one of four treatments (10/treatment): 1) natural photoperiod, 2) natural photoperiod + bST (14 mg/d Trobest i.m.), 3) long daily photoperiod (18 h of light and 6 h of darkness/d), or 4) long daily photoperiod + bST. Long daily photoperiod increased fat corrected milk (FCM) yield 1.9 kg/d versus natural photoperiod. Treatment with bST increased FCM 5.7 kg/d versus natural photoperiod, and long daily photoperiod + bST increased FCM 5.8 kg/d versus long daily photoperiod. Long daily photoperiod + bST was additive, increasing FCM 7.7 kg/d versus natural photoperiod. Serum somatotropin increased with bST, but not photoperiod, and bST increased serum insulin-like growth factor-I. Long daily photoperiod tended to increase prolactin; bST had no effect. Long daily photoperiod + bST increased dry matter intake (DMI) relative to natural photoperiod and natural photoperiod + bST; long daily photoperiod increased DMI relative to natural photoperiod + bST. Photoperiod had no effect on net energy balance; however, bST decreased net energy balance. Generally, body weight and milk composition did not differ among treatments. In conclusion, combination of bST with long-daily photoperiod tended to amplify the increases in milk yield observed with either treatment individually. PMID- 10480098 TI - Effects of secretion removal on bovine mammary gland function following an extended milk stasis. AB - The objective of this study was to determine whether lactation function could be reinitiated after a period of extended milk stasis. Involution was induced by milk stasis in lactating Holstein cows for a period of 11 d. On d 11, one side of the mammary gland was milked twice daily for 3 d. The contralateral side remained unmilked for the 14-d experimental period. Cows were slaughtered, and mammary tissue was collected from both udder halves for further analysis. Mammary secretion volume was partially restored in the milked udder half, but reestablished milk yields were variable among cows. A partial recovery of lactation function was further indicated by elevated levels of lactose and protein profiles resembling milk in mammary secretions from the milked glands. Lactose and protein profiles from the unmilked glands were similar to those of glands undergoing involution. Lactoferrin levels were elevated in secretions from the milked and unmilked udder halves. Casein and lactoferrin synthesis by mammary explants and beta-casein and lactoferrin mRNA abundance in mammary tissues corresponded to protein profiles from milked and unmilked mammary secretions. alpha-Lactalbumin mRNA was variable but was more abundant in the milked glands compared with the unmilked glands. Lectin fluorescence microscopy for soybean agglutinin preferentially stained the apical surface of the mammary epithelial cells from the milked glands. Staining was absent in the unmilked glands and suggested resumption of lactation function in all such milked glands. These results suggest that mammary involution can be partially reversible after 11 d of milk stasis. PMID- 10480099 TI - Effects of energy balance of hormones, ovarian activity, and recovered oocytes in lactating Holstein cows using transvaginal follicular aspiration. AB - The effects of energy balance on hormonal secretion patterns and the structure of recovered oocytes were evaluated in 20 lactating Holstein cows during two trial periods. Cows were randomly assigned to one of two dietary treatments formulated so that dry matter consumption was 3.6% of body weight (high energy; 1.78 Mcal/kg) or 3.2% of body weight (low energy; 1.52 Mcal/kg). Ovum recovery procedures were conducted twice weekly between d 30 and 100 of lactation. Follicle size and number were recorded. Follicular fluid aspirated from the largest follicle and serum samples were collected for hormone assay. Milk yield averaged 41.6 +/- 0.3 kg/d (mean +/- SE) for high energy fed cows and 32.8 +/- 0.3 kg/d for low energy fed cows. Oocyte numbers increased linearly from d 30 to 100 postpartum. Cows fed high energy diets produced more good (+) oocytes than did cows fed low energy diets. PMID- 10480100 TI - Short communication: an electronic probe versus milk progesterone as aids for reproductive management of small dairy herds. AB - A simple probe especially designed to take electrical resistance measurements at different positions in the anterior vagina of a cow was compared with milk progesterone determinations on 108 cows. Milk samples were taken 3 x weekly, 21 to 60 d postpartum, at the time of insemination, and 21 to 23 d later. Electrical resistance measurements were made on a similar schedule. In 10 other herds, 187 cows had only milk samples taken. No cows with high milk progesterone values became pregnant when inseminated, but the electrical resistance values were less accurate in designating which cows were suitable or unsuitable for insemination. Both low milk progesterone and low electrical resistance values 21 to 23 d after insemination provided an early and accurate indication of a need for reinsemination. These indicators were consistent with 94 to 100% of these cows being diagnosed as not pregnant 6 to 8 wk later. Daily probing, starting about 19 d after a previous insemination, could serve as an early check of pregnancy and assist in identifying cows for immediate reinsemination. PMID- 10480101 TI - The effects of accelerated growth rates and estrogen implants in prepubertal Holstein heifers on growth, feed efficiency, and blood parameters. AB - Sixty-eight Holstein heifers were used to determine the effects of accelerated growth rates by increased nutrient intake and estrogen implants on feed efficiency, structural growth, and blood parameters in heifers between 19 and 39 wk of age. At the beginning of the treatment period, the heifers were assigned to one of four treatment groups by using a randomized complete block design in a 2 x 2 factorial arrangement. The treatments were standard growth rate (700 g/d), accelerated growth rate (1000 g/d), standard growth rate with an estradiol implant, and accelerated growth rate with an estradiol implant. All heifers received the same diet, but dry matter intake was adjusted weekly to achieve the target rate of gain. Accelerating heifer growth rates from 705 to 1007 g/d improved feed efficiency 5.1%, increased the rate of withers height, heart girth, and hip width growth 12, 27, and 27%, respectively, and body condition scores 0.25 points. Estradiol implants improved feed efficiency 2.4% and decreased the rate of withers height 6% and heart girth growth 3.5%. Increased nutrient intake and average daily gain depressed mean plasma growth hormone and urea nitrogen content 17 and 7%, respectively, while elevating insulin-like growth factor-1 levels by 10%. Estradiol implants increased mean plasma growth hormone content by 29% and insulin-like growth factor-1 levels by 17%, but decreased urea nitrogen content by 11%. Feeding prepubertal heifers for accelerated growth rates increased structural growth with a small increase in body condition, whereas estradiol implants improved feed efficiency and decreased the growth rate of withers height and heart girth without affecting the rate of hip width growth. PMID- 10480102 TI - The effects of accelerated growth rates and estrogen implants in prepubertal Holstein heifers on estimates of mammary development and subsequent reproduction and milk production. AB - The objectives of this study were to determine the effects of accelerating growth rates and of imposing estrogen implants in prepubertal heifers on mammary development and subsequent reproduction and milk production. Sixty-eight Holstein heifers were assigned to 1 of 4 treatment groups by using a randomized complete block design in a 2 x 2 factorial arrangement. The treatments were standard growth rate (700 g/d) or accelerated growth rate (1000 g/d) and estrogen implant or no estrogen implant. The treatments were imposed over 20 wk, beginning at 4.5 mo of age and 130 kg of body weight (BW). During the treatment period, all heifers were fed individually and received the same diet, but the dry matter intake of each heifer was adjusted weekly to achieve the designated growth rate. The estrogen implants were removed at the end of the treatment period at 9.5 mo of age. After the treatment period, the heifers were group fed according to BW and age to allow the heifers to have a similar BW and age at calving. The accelerated growth regimen decreased age at puberty by 32 d. Age, BW, and body condition scores at calving were not significantly different among treatments. The accelerated prepubertal growth regimen decreased first lactation fat corrected milk yield 7.1%. Prepubertal heifers given estrogen implants produced 5.2% less fat-corrected milk during first lactation than did heifers not implanted with estrogen. Estrogen implants stimulated a large increase in teat length growth during the treatment period, but the advantage was lost posttreatment. Over both the treatment and posttreatment periods, the estrogen implants reduced teat length growth by 30%. Accelerated growth rates from 700 to 1000 g/d and estrogen implants in prepubertal heifers decreased first lactation milk production. PMID- 10480103 TI - Improving energy supply to late gestation and early postpartum dairy cows. AB - Sixty-five multiparous Holstein cows were used to test the effects of feeding diets of varied ruminal carbohydrate availability during the transition period on dry matter intake, blood metabolites, and lactational performance. Cows received total mixed rations containing either cracked corn or steam-flaked corn beginning 28 d prior to expected calving date. At parturition, cows were assigned to a postpartum total mixed ration that contained either cracked corn or steam-flacked corn. Diets were fed until 63 d in milk. No treatment effects on prepartum or postpartum dry matter intake, body weight, and body condition score were observed. Cows fed steam-flaked corn had lower blood urea N concentrations during the prepartum period and lower plasma nonesterified fatty acid concentrations during the prepartum and postpartum periods. Cows fed steam-flaked corn postpartum produced 2.3 kg/d more milk than cows fed cracked corn during the first 63 d in milk. Fat corrected milk showed no treatment effect. Seven cows were used to evaluate treatment effects on ruminal fermentation and digesta kinetics. Prepartum and postpartum treatments had minimal effects on ruminal fermentation. Feeding steam-flaked corn prepartum decreased apparent fiber digestibility and ruminal NH3 N. Feeding steam-flaked corn postpartum decreased the acetate to propionate ratio. Prepartum and postpartum treatments did not affect digesta kinetics. An increase in ruminal carbohydrate availability during the postpartum period enhanced milk production, but had variable results on ruminal fermentation. PMID- 10480104 TI - Synchronization of carbohydrate and protein sources on fermentation and passage rates in dairy cows. AB - Four ruminally cannulated Holstein cows in midlactation were randomly assigned to a 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments to evaluate two nonstructural carbohydrate sources (corn or barley) with two sources of ruminally undegradable protein (soybean meal or extruded soybean meal) on milk production, ruminal fermentation, and digesta passage rates. Milk production (25.1, 27.5, 23.8, and 23.5 kg/d for the corn and soybean meal, corn and extruded soybean meal, barley and soybean meal, and barley and extruded soybean meal, respectively) and dry matter intake per unit of body weight (3.9, 4.1, 3.7, and 3.7%) were greater for cows fed corn than for cows fed barley and were similar for cows fed soybean meal or extruded soybean meal. Concentrations of ruminal NH3 N were greater for cows fed the corn and soybean meal diet than for cows fed other diets (15.0, 10.4, 9.0, and 11.3 mg/dl). Rumen volatile fatty acid concentrations were greater for cows fed corn than barley (133, 139, 121, and 118 mumol/ml). Fractional passage rates of solids from the rumen were greater for cows fed the barley and soybean meal diet than cows fed the corn and soybean meal diet (3.4, 3.9, 4.2, and 3.8%/h), and ruminal liquid dilution rates were similar for cows fed all diets (11.2, 11.0, 11.1, and 11.9%/h). The attempt to synchronize ruminal nonstructural carbohydrate and crude protein degradability produced minimal benefits for midlactation dairy cows. PMID- 10480105 TI - Comparison of methods of in vitro dry matter digestibility for ten feeds. AB - The objectives were to determine if the in vitro dry matter (DM) digestibility was similar by using a traditional method compared to the new DAISYII system and to determine if in vitro DM digestibility was similar for sources of inoculum from two different donor cow diets, all forage or total mixed ration. Ten feeds were digested by the traditional method, the DAISYII method with same feeds in a digestion vessel, and the DAISYII method with different feeds in a digestion vessel. The study used a 2 x 3 factorial arrangement of treatments with two sources of inoculum and three methods. The study was replicated. Source of inoculum affected in vitro DM digestibility; the grass hay donor cow diet resulted in lower digestibility values in six of the 10 feeds tested. Method did not significantly affect the digestibility values, and there were no significant source by method interactions. The DAISYII method can be used to increase labor efficiency in the in vitro DM digestibility analysis, and forages and grains can be analyzed together in a single digestion vessel. PMID- 10480106 TI - Optimized dairy grazing systems in the northeast United States and New Zealand. I. Model description and evaluation. AB - Parallels exist in the recent developments of dairy systems in the Northeast United States and New Zealand because of greater use of pasture grazing and feed supplements, respectively. Lessons can be learned from each system. However, major differences exist between the regions in the patterns of pasture production, the costs of supplementary feed, and milk prices. These differences affect the optimum use of feed. In this paper, a linear programming model developed to determine optimum feeding strategies for dairy systems in each country is presented. The model optimizes grazing management (rotation lengths) and the conservation of pasture subject to constraints on their use. Other feed resources include N fertilizer, grain, corn silage, and alfalfa silage. All feeds are represented in energy terms. The substitution of pasture intake by grain and forage supplements is included, and cow performance can be optimized by choosing from 73 seasonal calving herds that vary in calving date, lactation length, and daily milk production. The model predicts that marginal responses to grain feeding are between 1.35 and 1.8 kg of milk/kg of grain dry matter supplement, well within the range of responses reported in the literature. Evaluation of the model against data from nine grazing system treatments in New Zealand and two in Pennsylvania showed that model predictions averaged +3% (New Zealand) and +0.04% (Northeast) of measured milk production. The model could be used with confidence to study systems in both the Northeast United States and New Zealand. PMID- 10480107 TI - Optimized dairy grazing systems in the northeast United States and New Zealand. II. System analysis. AB - Factors that optimize milk production from Northeast United States and New Zealand grazing systems are compared using a linear programming model. The objective function maximized gross margin per hectare of land farmed. The experimental design compared the optimum characteristics of each system over a range of milk prices. The Northeast has a shorter grazing season and lower cropping costs than New Zealand. The optimum pasture area was 49% of the farm for Northeast systems. Gross margins declined rapidly above 55% or below 36% pasture area. The optimum stocking rate was 1.13 cows/ha, or 2.3 cows/ha of pasture. Optimum per cow production was higher for Northeast [7105 kg of fat-corrected milk (FCM)] than New Zealand (5710 kg of FCM) systems. This was related to lower grain relative to milk prices in the Northeast. New Zealand, all-pasture systems gave the lowest cost per unit of milk but also gave the lowest gross margin across all milk price scenarios. The best use of purchased feed in New Zealand systems was to support increased stocking rate rather than per cow production. Optimum grazing management practices were similar for supplemented New Zealand and Northeast systems. All-pasture New Zealand systems are characterized by short lactations and long autumn rotations to transfer pasture in situ for winter feeding. Higher costs per unit of milk produced will be an inevitable consequence of maximizing gross margin at high milk prices in New Zealand systems. PMID- 10480108 TI - Effects of lasalocid on performance of lactating dairy cows. AB - Sixty lactating dairy cows (30 multiparous and 30 primiparous) were used in a completely randomized block design to determine the effect of lasalocid supplementation on dairy cow performance. Starting wk 2 prepartum and lasting through wk 17 of lactation, cows received one of three experimental diets. The experimental total mixed rations were control (CD), control + 10 mg/kg of lasalocid (CD + 10) and control + 20 mg/kg of lasalocid (CD + 20). The alfalfa based control diet (40:60; forage:concentrate) was formulated to contain 18% crude protein, 35% nonstructural carbohydrates, 31% neutral detergent fiber, and 6.6% ether extract. Lasalocid supplementation linearly decreased dry matter intake (DMI) without affecting milk production or milk composition. Mean milk production and percentages of fat and protein were 30.0, 30.8, and 28.6; 3.56, 3.51, and 3.63; 3.06, 3.05, and 3.09; respectively for treatments CD, CD + 10, and CD + 20. Lasalocid supplementation decreased milk urea N (MUN) when compared to control cows, and increasing supplementation caused a significant linear decrease in MUN. For the primiparous cows, lasalocid supplementation decreased DMI and MUN while increasing body condition score and feed efficiency. Results from this study indicate that lactating dairy cows and, in particular, primiparous cows may benefit from lasalocid supplementation in terms of more efficient utilization of nutrients for milk production, reduced MUN levels, reduced body condition loss, and higher margin over feed cost. PMID- 10480109 TI - Effects of intrinsic potassium in artificially dried grass and supplemental potassium bicarbonate on apparent magnesium absorption in dry cows. AB - Literature data indicate that the form of K in the ration can affect its inhibitory influence on Mg absorption in ruminants. We tested whether identical amounts of K either intrinsically present in artificially dried grass or present in added KHCO3 have different effects on Mg absorption in dry cows. In a 3 x 3 Latin square design, six cows were fed rations consisting of low-K grass and concentrate with or without KHCO3 or a ration consisting of high-K grass with concentrate without added KHCO3. Each ration was given for a period of 4 wk. The ration low in intrinsic K contained 26 g of K/kg of dry matter, the ration low in intrinsic K plus KHCO3 contained 43 g of K/kg of dry matter, and the ration high in intrinsic K also contained 43 g of K/kg of dry matter. The three rations were balanced for crude protein, crude fat, crude fiber, Mg (2.2 g/kg of dry matter), Ca, P, and Na. Apparent Mg absorption was 10.8 +/- 1.54% of intake (mean +/- SE, n = 6) when the cows were fed the low-K ration, but dropped to 1.9 +/- 3.4 and 2.1 +/- 1.9% of intake, respectively, when the rations high in KHCO3 and high in intrinsic K were fed. The two high-K rations induced similar increases in ruminal K concentrations both before and after feed consumption. The feeding of KHCO3 did not influence ruminal pH. The intake of extra K may raise ruminal K concentrations, which increases the transmural potential difference so that Mg transport across the rumen epithelium becomes depressed. Thus, intrinsic and added K had identical effects on ruminal K concentrations and on Mg absorption. Feeding trials with ruminants in which K intakes are manipulated with the use of KHCO3 may reflect those cases when concentrations of K intrinsically present in feedstuffs may vary. PMID- 10480110 TI - Technical note: forms and route of vitamin C supplementation for cows. AB - The purpose of this study was to compare the effect of multiple oral supplementation of two forms of vitamin C on plasma ascorbic acid concentrations in dairy cows. Vitamin C was administrated at the rate of 40 g/d during a 5-d period. The supplements were vitamin C in the form of a fine powder and vitamin C coated with ethyl cellulose. In addition, another group of cows provided with vitamin C in the form of fine powder through an abomasal fistula was enrolled in the study. The effect of vitamin C supplementation was assessed by the change in plasma ascorbic acid concentrations during a 5-d period of vitamin C supplementation. A tendency existed for cows that were dosed with vitamin C in the abomasum to have higher ascorbic acid than cows supplemented orally. In the abomasum-dosed cows, plasma ascorbic acid value varied between 3.6 and 4.4 micrograms/ml; the maximum ascorbic acid content was 4.4 micrograms/ml at 54 h. Cows dosed orally with vitamin C coated with ethyl cellulose had higher plasma ascorbic acid concentrations than did cows that received vitamin C in the form of fine powder. In the former group, the maximum ascorbic acid concentration occurred at 30 h after dosing (5 micrograms/ml). PMID- 10480111 TI - Genetic analysis of fertility in dairy cattle using negative binomial mixed models. AB - Two negative binomial mixed models with different dispersion specifications were compared for analysis of dairy reproduction count data. The first model was developed previously and had heterogeneous overdispersion in an associated logarithmic scale, assigning greater uncertainty to observations with smaller conditional expectations. The second model postulated homogeneous overdispersion across all data. A simulation study was used to compare marginal modal estimates of additive genetic variance, based on these two negative binomial models, with analogous estimates computed by an overdispersed Poisson mixed model. Estimators from the second negative binomial and overdispersed Poisson models had better frequentist properties than did those from the first negative binomial model. Nevertheless, application to a data set of number of artificial inseminations until conception in Holstein heifers suggested a slightly better fit of the first negative binomial model. A marginal likelihood ratio test indicated that the additive genetic variance was significant. Cross-validation analyses suggested that the two negative binomial mixed models had slightly better predictive ability than a linear mixed model. PMID- 10480112 TI - Seasonal incidence and geographical variation of Nebraska mosquitoes, 1994-95. AB - A mosquito survey was conducted in Nebraska during 1994-95. Mosquitoes were collected by CO2-baited CDC light traps at 11 localities throughout Nebraska. Total mosquitoes collected was 685,581 per trap (mean = 790) and 1,163,741 per trap (mean = 1,372) during 1994 and 1995, respectively. Twenty-seven mosquito species representing 8 genera were collected. The most abundant mosquitoes collected statewide were Aedes vexans at 74% of the 2-year total, Culex tarsalis at 11%; Aedes trivittatus, 5%; Aedes melanimon, 1.5%; and Aedes triseriatus, 1%. The floodwater mosquitoes were more prevalent in eastern Nebraska while standing water mosquitoes, especially Culex tarsalis, were more prevalent in the west. PMID- 10480113 TI - Culex (Micraedes) biscaynensis n. sp. from Florida (Diptera: Culicidae). AB - The female, male, male genitalia, pupa, and larva of Culex biscaynensis, a new species in the Bisulcatus Group of subgenus Micraedes, are described and illustrated. The species, which is presently known only from south Florida, United States, is most similar to Culex (Micraedes) antillummagnorum Dyar. PMID- 10480114 TI - Mosquito immatures in drought-prone and drought-resistant bamboo stumps in Flores, Indonesia. AB - Mosquito species in water-filled bamboo stumps were compared with those emerged from dormant eggs in dry stumps. Tripteroides spp. was more abundant than Aedes (Stegomyia) spp. (mainly annandalei and albopictus) in water-filled stumps, whereas the latter was more abundant than the former in dry stumps. During 5 days after inundation, Aedes (Stegomyia) spp. developed faster than Tripteroides spp. Density of Tripteroides spp. was high in narrow and deep stumps. Both taxa of mosquitoes were at low densities in water-filled stumps with large detritivores (Tipulidae and Oligochaeta). PMID- 10480115 TI - Comparison of two American biophysics mosquito traps: the professional and a new counterflow geometry trap. AB - Large cage and field studies were conducted to compare the efficacy of 2 American Biophysics Corporation mosquito traps, the standard professional (PRO) trap and a new counterflow geometry (CFG) trap. The PRO trap utilizes conventional downdraft technology and the CFG trap uses a patent-pending technology. In large cage studies, similarly baited CFG traps captured approximately 1.7 times as many laboratory-reared Aedes taeniorhynchus as the PRO trap. The CFG trap baited with CO2 + octenol resulted in significantly reduced landing counts compared to all other treatments; mean landing count was reduced from 233.8 (12.99/min), when no trap was present, to 24.7 (1.37/min). In field studies against natural populations of woodland species, the CFG trap captured 7.8 times more mosquitoes than the PRO trap overall, and approximately 11 times more Anopheles crucians, Anopheles quadrimaculatus, and Culex erraticus. PMID- 10480116 TI - Population dynamics of Anopheles culicifacies and malaria in the tribal area of central India. AB - A longitudinal study (1993-94) on malaria was conducted in Dungaria, a typical forest fringe tribal village in Mandla district of central India (Madhya Pradesh). Our initial objective was to obtain in-depth baseline data on malaria transmission in the tribal village to elucidate the factors responsible for persistent malaria in the area and thereby to help in formulating an improved malaria control program. Anopheles culicifacies Giles was the predominant vector of malaria, although Anopheles fluviatilis James were recorded in small numbers. The transmission season was from May to November. Analysis of the malaria cases revealed hyperendemic malaria, with Plasmodium falciparum the predominant species. The prevalence of Plasmodium vivax was mainly in the summer and that of P. falciparum in autumn. The study suggested that a number of factors were responsible for the continuation of malaria transmission in the village. PMID- 10480117 TI - Oviposition preference for freshwater in the coastal malaria vector, Anopheles farauti. AB - Oviposition preference of the Australasian coastal malaria vector Anopheles farauti s.s. for water of varying salinity was determined in the laboratory to help understand the distribution and control of this species in the field. Numbers of eggs laid showed an inverse relationship with salinity; of 5 NaCl concentrations most eggs were laid in distilled water but some were laid in 3.17% NaCl (the salinity of seawater). The association of An. farauti with coastal areas occurs in spite of an aversion to salt water by ovipositing females. Factors other than salinity must be the primary determinants of distribution. Increasing the salinity of larval habitats will not totally prevent An. farauti from laying eggs. Elimination of this species may not occur unless salinity is kept high enough to prevent complete larval development. PMID- 10480118 TI - Potential for Central American mosquitoes to transmit epizootic and enzootic strains of Venezuelan equine encephalitis virus. AB - Experimental studies were undertaken to compare the vector competence of Culex (Melanoconion) taeniopus Dyar and Knab, Culex (Melanoconion) ocossa Dyar and Knab, and Psorophora confinnis (Lynch Arribalzalga) from Central America for epizootic (IAB) and enzootic (IE) strains of Venezuelan equine encephalitis (VEE) virus. Virus infection and dissemination rates were significantly higher in Cx. taeniopus orally exposed to IE as compared to those orally exposed to IAB virus. In contrast, both infection and dissemination rates were similar in Cx. ocossa exposed to either IAB or IE strains of VEE virus. Thus, susceptibility to epizootic and enzootic strains of VEE virus seems to be species specific within the subgenus Culex (Melanoconion). Both species transmitted each strain of VEE virus after intrathoracic inoculation, indicating that a midgut barrier affected vector competence in these species. Psorophora confinnis was equally susceptible to both IAB and IE viruses, but apparently had a salivary gland barrier, as only 1 of 16 mosquitoes with a disseminated infection transmitted VEE virus by bite. PMID- 10480119 TI - A geographical information system to manage mosquito and arbovirus surveillance and control data in the Coachella Valley of California. AB - A geographical information system was developed to monitor and analyze mosquito abundance and encephalitis virus activity in the Coachella Valley to facilitate mosquito control operations. Data layers include soil types, vegetation types, irrigation method, standpipes, larval occurrence, adult abundance, and viral transmission to sentinel chickens. Base maps are digitized aerial photographs, with data entry done through sections of the range/township system. The image resolution of a section is 100 ft2 (9.3 m2) per pixel. This system currently is operational and in use by the Coachella Valley Mosquito and Vector Control District for data management. PMID- 10480120 TI - Resistance to two pyrethroids in Anopheles sinensis from Zhejiang, China. AB - Probabilities of pyrethroid resistant genotypes in natural populations of Anopheles sinensis Wiedemann were measured with deltamethrin and permethrin. The median lethal concentrations (LC50s) of deltamethrin and permethrin in the susceptible larval population were 0.0209 and 0.1747 ppm, respectively. Under dosages that produced 99% mortality in susceptible laboratory strains of larvae, the lethal percentage of Cangnan larval field populations after 20 min of exposure was only 61.23% for deltamethrin and 64.92% for permethrin. This was much lower than those of other natural populations. Also, the probability of pyrethroid-resistant genotypes in Cangnan adult field populations was at the highest, reaching 0.5867. The results are discussed in relation to future mosquito control programs. PMID- 10480121 TI - Comparison of electrostatic versus nonelectrostatic ULV sprays of Aqua Reslin against Anopheles quadrimaculatus adults. AB - An electrostatic truck-mounted spray system (Spectrum, Houston, TX) and a nonelectrostatic spray system (Micro-Gen G-4, San Antonio, TX) were tested to determine the feasibility of electrostatically charging Aqua Reslin, a water based permethrin insecticide, and ascertain whether an electrostatic charge would increase the efficiency of Aqua Reslin against Anopheles quadrimaculatus adults. Parameters tested for both machines included mean mass median diameter (MMD) of droplets, number of drops per cm2, and posttreatment percent mortality at 1, 12, and 24 h. Results indicated that the electrostatically charged droplets produced greater mortality at each distance and hour posttreatment. Correlation coefficients and linear equations were calculated for distance-mortality, MMD mortality, drops per cm2-mortality, distance-drops per cm2, distance-MMD, and MMD drops per cm2. Results indicated that the electrostatic drops demonstrated strong correlations between each paired variable, whereas the nonelectrostatic drops showed poor correlation between drops per cm2-mortality, distance-drops per cm2, and MMD-drops per cm2. However, from this trial, these differences cannot be attributed purely to the electrostatic charge because significant differences in droplet size can affect spray performance. PMID- 10480122 TI - Selection and characterization of temephos resistance in a population of Aedes aegypti from Tortola, British Virgin Islands. AB - A collection of Aedes aegypti from Tortola, British Virgin Islands, with a high level of temephos resistance (46.8-fold at the 95% lethal concentration [LC95]) was selected to higher resistance with temephos in the laboratory. After 13 generations of pressure, the temephos resistance ratio increased to 180.6 (LC95), whereas in the absence of selection pressure the resistance ratio declined to 8.5. Relatively low levels of resistance or cross-resistance to other organophosphate and carbamate insecticides, and a high level of resistance to the pyrethroid permethrin were also observed. Synergism tests implicated detoxifying esterases in temephos resistance and the presence of elevated esterase activity was confirmed by biochemical tests; however, no evidence was found of insensitive acetylcholinesterase. Mendelian crosses indicated that temephos resistance was inherited as a monofactorial trait. The presence of high levels of temephos and permethrin resistance in Ae. aegypti has important implications for Aedes control programs. PMID- 10480123 TI - Effects of temephos (Abate 4E) on fiddler crabs (Uca pugnax and Uca minax) on a Delaware salt marsh. AB - The nontarget effects of temephos (as Abate 4E, 44.6% active ingredient) on fiddler crabs were examined on the salt marsh at Bombay Hook National Wildlife Refuge, near Dover, DE. Six 170 x 170-m plots were established; 3 were sprayed on 4 occasions at a rate of 1.5 fl oz/acre (0.054 kg active ingredient/ha) and 3 were controls. On each plot, marsh fiddler crab (Uca pugnax) populations were monitored by repeatedly counting the number of burrow holes in 2 counting areas marked out along tidal guts. One half of each counting area was covered with bird netting to evaluate sublethal toxic effects, which, if present, could result in increased susceptibility to bird predation. A statistically significant linear association was established between the number of holes and the number of crabs. No significant differences were found in the numbers of holes (or crabs) in the sprayed vs. control plots and in the covered vs. uncovered sections. However, survival of juvenile crabs in in situ bioassays was significantly reduced (16% lower) by the spraying. Median acetylcholinesterase activity in claw muscle of red-jointed fiddler crabs (Uca minax) collected 2 days after an operational spray with Abate 4E was significantly reduced (28% lower) compared to unsprayed crabs. In view of the toxicity to juvenile crabs and the cholinesterase inhibition, we recommend continued monitoring and research for nontarget impacts of Abate 4E on fiddler crabs to establish whether the reported level of cholinesterase inhibition results in acute or chronic toxicity. PMID- 10480124 TI - Effects of ultra-low volume pyrethrin, malathion, and permethrin on nontarget invertebrates, sentinel mosquitoes, and mosquitofish in seasonally impounded wetlands. AB - Wildlife managers are concerned that insecticides used to control mosquitoes could suppress invertebrates on which wildlife feed. We assessed whether ultra low volume (ULV) applications of pyrethrin, permethrin, and malathion for control of adult mosquitoes reduced macroinvertebrate abundance and biomass or killed mosquitofish in seasonal wetlands in California. Pyrethrin was applied over 3 seasonal wetlands on Sutter National Wildlife Refuge (NWR), and malathion or permethrin were each applied over 2 seasonal wetlands on the Colusa NWR. Three control wetlands were used per site. We measured aquatic macroinvertebrate abundance and biomass before and after insecticide application and compared the survival of mosquito larvae held in sentinel cages. At Colusa, we also used mosquitofish as sentinels, caged adult mosquitoes over the wetlands to test for pesticide efficacy and drift, and sampled night-flying insects using ultraviolet light traps. Results showed no detectable reductions in the abundance or biomass of aquatic macroinvertebrates in treated wetlands. Larval mosquitoes showed high survival in all areas. All adult mosquitoes died when caged over wetlands treated with malathion or permethrin, but all survived in controls. All mosquitofish survived. Flying insect abundance decreased after insecticide application in both treated and control wetlands but rebounded in 48 h. Results indicated that ULV applications of these insecticides to control adult mosquitoes are unlikely to have substantial effects on the aquatic insects or fish in seasonal wetlands. PMID- 10480125 TI - Field evaluation of the repellents deet, CIC-4, and AI3-37220 against Anopheles in Lae, Papua New Guinea. AB - The repellents diethylmethylbenzamide (deet), (2-hydroxymethylcyclohexyl) acetic acid lactone (CIC-4), and 1-(3-cyclohexen-1-yl-carbonyl)-2-methylpiperidine (AI3 37220) were compared for their effectiveness in protecting 5 soldiers against the bites of Anopheles spp. at a village in Papua New Guinea. All 3 repellents, applied as 25% ethanol concentrations, provided > or = 95% protection against primarily An. farauti 4 for at least 3 h after application. PMID- 10480127 TI - Variation in the protection periods of repellents on individual human subjects: an analytical review. AB - Mosquito repellent test data from the literature were analyzed to estimate mean protection periods and among-subjects standard deviations. Standard deviations were a linear function of the means. Numbers of subjects needed to determine mean protection periods of 1-8 h with confidence limits of +/- 0.5, 1.0, 1.5, and 2.0 h at the 99 and 95% levels of confidence were computed from regression values of the standard deviation, and a table of sample sizes was constructed for use in planning repellent tests. PMID- 10480126 TI - Laboratory evaluation of AI3-37220, AI3-35765, CIC-4, and deet repellents against three species of mosquitoes. AB - Four repellents, N,N-diethyl-3-methyl-benzamide (deet), 2-hydroxy-methyl cyclohexyl acetic acid lactone (CIC-4), and 2 piperidines (1-[3-cyclohexen-1 ylcarbonyl] piperidine [AI3-35765] and 1-[3-cyclohexen-1-ylcarbonyl]-2 methylpiperidine [AI3-37220]) were evaluated alone and in combination against Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus using a modified in vitro test system. This method was a valuable tool for comparing effective concentrations of the new compounds. Because of the controlled conditions of the test, it was possible to use the results of assays that had been conducted over a 5-year period and to perform the many replications necessary to evaluate combinations of compounds. The new candidate repellents were generally as effective as deet. Although speculative at this time, there was some evidence of synergistic interaction. Repellent combinations of CIC-4/AI3-37220/AI3-35767, deet/AI3-35765, and deet/AI3-37220/AI3-35765 against An. stephensi and CIC-4/AI3 35765, deet/AI3-37220/AI3-35765, AI3-37220/AI3-35765, and CIC-4/AI3-37220 against Ae. aegypti were more effective than the component compounds alone. PMID- 10480128 TI - Field evaluation of new water-dispersible granular formulations of Bacillus thuringiensis ssp. israelensis and Bacillus sphaericus against Culex mosquitoes in microcosms. AB - A variety of formulations of Bacillus thuringiensis var. israelensis de Barjac (B.t.i.) and Bacillus sphaericus Neide (B.s.) have been studied for mosquito control under laboratory and field conditions. High efficacy, specificity, low risk of development of resistance, long shelf-life, and transportability, as well as the safety to nontarget organisms of these 2 microbial agents have been well documented. Some of the currently available formulations of B.t.i. and B.s. have low potency per unit mass. Research and development efforts are focusing on commercializing formulations with high potency and low minimum effective dosage that are suitable for long-distance shipment. To achieve this goal, new water dispersible granule (WDG) formulations of both microbial agents were prepared and made available by Abbott Laboratories for evaluation. The newly developed WDGs of B.t.i. and B.s. with high potency dispersed readily in water with gentle agitation. These WDGs were evaluated and the minimum effective dosages were determined in microcosms against natural populations of Culex mosquitoes. The minimum effective dosage for B.t.i. WDGs with 4,000 International Toxic Units (ITU)/mg was 0.27-0.53 lb/acre which yielded significant control for up to 7-12 days. The minimum effective dosage for B.s. WDGs with 350-630 ITU/mg was 0.05 0.10 lb/acre, which yielded significant control of immature mosquitoes for up to 14-20 days. PMID- 10480129 TI - Duration of activity of the microbial larvicide VectoLex CG (Bacillus sphaericus) in Illinois catch basins and waste tires. AB - The duration of activity of a formulation of Bacillus sphaericus, VectoLex CG, for control of Culex species was evaluated in 338 catch basins in Urbana, IL, and compared to Altosid in 346 catch basins in Champaign, IL. The activity of VectoLex in car and truck waste tires was evaluated in a tire dump located in Pembroke Township, IL. In catch basins, 1 g of VectoLex per catch basin gave the same control as one Altosid briquet. Both larvicides were effective against Culex sp. in catch basins for 1 month, and the duration of control with VectoLex lasted 44 days in one catch basin. VectoLex was considerably cheaper to apply than Altosid briquets, at 0.64 cents per catch basin compared to 90.75 cents, respectively. However, the Altosid briquets were judged to be easier to apply from a vehicle than VectoLex granules. VectoLex (22.6 kg) was used to treat approximately 6,000 car and truck tires; some of the tires were in direct sunlight whereas others were shaded. Aedes triseriatus was the dominant species in these tires. Tires treated with VectoLex contained significantly fewer mosquitoes than control tires, and even 65 days after application, control tires were 16.7 times more likely to contain larvae. We conclude that VectoLex was effective when used in Illinois catch basins and tire dumps, and emphasize that it is more appropriate to base tire treatment rates on the total number of tires present than on a kilogram per hectare basis. PMID- 10480130 TI - Residual activity of Bacillus thuringiensis serovars medellin and jegathesan on Culex pipiens and Aedes aegypti larvae. AB - Bacillus thuringiensis serovar medellin strain 163-131 and Bacillus thuringiensis serovar jegathesan (B.t.jeg.) strain 367 are very toxic to mosquito larvae. However, they are 10 times less toxic than Bacillus thuringiensis var. israelensis (B.t.i.) to mosquito larvae under laboratory conditions. Lyophilized powders were produced from these strains and their toxicities were compared to that of powder produced from the B.t.i. strain. Larvicidal activity was titrated using Aedes aegypti (Bora-Bora strain) larvae, with IPS82 powder as the standard. The efficacy of these powders in the field was determined using Culex pipiens (Montpellier strain) in Paris, France, and Ae. aegypti larvae (French Guiana strain) in Cayenne, French Guiana, in standardized conditions. Residual activity was also assessed in the laboratory, using Cx. pipiens (SLAB strain), in Montpellier, France. Any negative effect of direct sunlight, soil, or polluted water on the residual activity of the 3 powders was recorded. Increasing bacterial concentration by a factor of 8 had little effect on the duration of larvicidal activity, except in the presence of polluted water and when substrate was added. All powders had similar initial efficacies against both types of mosquito larvae, in all conditions except water rich in organic matter. Bacillus thuringiensis serovar medellin had the lowest residual activity, both in the laboratory and in the field, whereas B.t.jeg. remained toxic for as long as B.t.i. PMID- 10480131 TI - Comparative efficacy of the threespine stickleback (Gasterosteus aculeatus) and the mosquitofish (Gambusia affinis) for mosquito control. AB - The effectiveness of the threespine stickleback as a mosquito control agent was compared to that of the mosquitofish in 28-m2 earthen ponds during 26-wk experiments where the 2 fish were stocked alone and together. Relative to ponds without fish, the stickleback was not effective for controlling larval mosquito populations; however, sticklebacks reduced the abundance of Culex pupae. Mosquitofish provided significant levels of control whether stocked alone or concurrently with the stickleback. As compared to mosquitofish alone, mosquito control was not significantly enhanced when both fish were stocked together. Mortality of adult sticklebacks was related to a gradient of increasing water temperature across the ponds rather than the direct effects of other abiotic factors such as low dissolved oxygen concentrations or biotic interactions with the mosquitofish. The stickleback exhibited a lower thermal tolerance and slower population recruitment as compared to the mosquitofish populations,which reproduced successfully in water > 33 degrees C and grew rapidly. Stickleback biomass either declined or increased slightly (approximately 50% of initial stocking weight). Mosquitofish biomass increased 33- to 38-fold at rates averaging between 0.079 and 0.095 g wet weight/g/day and total wet weight per pond at 6 wk after stocking did not differ significantly between the 2 mosquitofish treatments. PMID- 10480132 TI - Seasonal distribution and relationship to temperature and precipitation of the most abundant species of Culicoides in five provinces of Andalusia, Spain. AB - A total of 177,344 Culicoides specimens were collected from 3,109 light trap collections made weekly from August 1990 to October 1991 at 62 sites in the provinces of Cadiz, Cordoba, Huelva, Seville, and Malaga, Spain. Reported for the 1st time are Culicoides agathensis, Culicoides bahrainensis, Culicoides marcleti, and Culicoides odiatus in the Iberian Peninsula, and Culicoides scoticus in Andalusia, Spain. As a group, Culicoides were active throughout the year. The 3 most common species were Culicoides imicola (56,254), Culicoides newsteadi (24,359), and Culicoides circumscriptus (16,720). Numbers of C. imicola peaked in October, C. newsteadi peaked in May, and C. circumscriptus peaked in June. Based on regression analyses, the optimal minimum and maximum air temperatures, respectively, for adult insect activity were approximately > or = 18 degrees C and > or = 38 degrees C for C. imicola, 12 degrees C for C. newsteadi, 14 degrees C for C. circumscriptus (minimum temperature only), 16 degrees C and > or = 38 degrees C for "other" Culicoides, and 14 degrees C and 32 degrees C for total Culicoides. Optimal minimum and maximum temperatures could not be determined for Culicoides pulicaris, Culicoides punctatus, subgenus Monoculicoides, and the Culicoides obsoletus group. During August and September, the months when African horse sickness outbreaks occurred, C. imicola was the predominant species in the coastal Mediterranean zone. If the "other" Culicoides spp. group was not considered, the predominant species were C. imicola in the Guadalquivir River valley zone, C. newsteadi in the Subbetica mountainous range zone, and subgenus Monoculicoides in the coastal Atlantic zone. PMID- 10480133 TI - Development of the first black fly (Diptera: Simuliidae) management program in Argentina and comparison with other programs. AB - In response to increasing pest populations of black flies, the government of Argentina initiated a pilot program to evaluate the effectiveness of larval control in the irrigation system of the Negro River Valley, an 18,240-km2 area in Patagonia. The extensive system of irrigation canals, drainage ditches, and natural waterways not only provides water for agriculture and general habitation but also affords habitat for immature black flies. Three species (Simulium bonaerense, Simulium wolffhuegeli, and Simulium nigristrigatum) are primary pests of humans and animals in this area. Trials were conducted using Vectobac AS in representative irrigation canals, a drainage ditch, and a medium-sized river. Most trials resulted in effective larval mortality and insecticide carry. Based on a comparison with 11 black fly suppression programs throughout the world, a suppression program for the Negro River Valley has a projected cost of approximately $1,623,360/year. This study represents the initial steps in development of the 1st areawide black fly suppression program in Argentina. PMID- 10480134 TI - First record of Aedes albopictus (Skuse) from Argentina. AB - Aedes albopictus was collected in San Antonio City, Misiones Province, Argentina, near the border with Brazil in March, 1998. Collected material included 12 pupae, 1 recently emerged adult female, and a few larvae, which were found in a dish, a plough disk, and a tin pot. From the collected pupae, 5 males and 4 females emerged in the laboratory. PMID- 10480136 TI - Current status of Aedes albopictus and Aedes atropalpus in Italy. AB - Eight years after the 1st record in Italy, scattered foci of Aedes albopictus are reported in 9 regions and 107 municipalities belonging to 22 provinces, mainly located in the northeastern part of the country (Veneto region). In almost all infested areas the species is well controlled and at low levels of density, through source reduction and antilarval treatments. Aedes atropalpus, 1st recorded in 1996, remains limited to the original focus in the province of Treviso. Surveillance and control of both species are carried out by local health agencies within a national program coordinated by Istituto Superiore di Sanita (national Institute of Public Health). PMID- 10480135 TI - First record of Toxorhynchites rutilus (Diptera: Culicidae) in Massachusetts. AB - The 1st recorded capture of Toxorhynchites rutilus in Massachusetts, USA, is reported. This capture represents the northernmost record of Tx. rutilus. The implications of this capture are discussed in the context of range expansion, evolution, and climatic change. PMID- 10480137 TI - [The effect of polymorphisms of the angiotensin-converting enzyme and angiotensinogen genes on the phenotypic expression of Spanish patients with hypertrophic cardiomyopathy]. AB - BACKGROUND: Hypertrophic cardiomyopathy (HCM) has a great variability in its morphofunctional expression. This study analyzes whether angiotensin-converting enzyme (ACE) and angiotensinogen (AGT) gene polymorphisms modulate the phenotypic expression in Spanish patients with HCM. PATIENTS AND METHODS: Forty Spanish HCM patients were studied. Twenty-six out of these 40 patients belonged to 7 families with familial HCM, and the remaining 14 patients had either a sporadic HCM or a HCM with unknown family incidence. A group of 269 healthy subjects was included as control for the genotype study. Maximal wall thickness, ventricular mass and several diastolic function indexes were measured in each patient by Doppler echocardiography. The insertion/deletion (I/D) polymorphism of ACE gene and the M235T polymorphism of AGT gene were studied in both patients and healthy subjects. RESULTS: A higher frequency in patients than in controls was found for D allele (0.79 vs 0.64; p = 0.02) and for DD genotype (62.5 vs 41.2%; p = 0.02). Conversely, no difference was observed in M235T polymorphism between both groups. Neither DD genotype of ACE, nor TT genotype of AGT determined a greater degree of ventricular hypertrophy or a worse diastolic function in patients with HCM. CONCLUSIONS: D allele and DD genotype are predisposing factors to express HCM. In this series of Spanish patients, I/D polymorphism of ACE and M235T polymorphism of AGT do not modify phenotypic expression of HCM. PMID- 10480138 TI - [The effect of the angiotensinogen M235T and the angiotensin-converting enzyme I/D polymorphisms on arterial hypertension and other cardiovascular risk factors]. AB - BACKGROUND: It has been reported the association between M235T angiotensinogen (AGT) and I/D angiotensin converting enzyme (ACE) gene polymorphisms and hypertension and other cardiovascular risk factors. However there are few data about Spanish population. So that we have studied the relationship among the aforementioned polymorphisms and hypertension and the possibility of association between any polymorphism and a worse cardiovascular risk profile. PATIENTS AND METHODS: 251 hypertensive and 245 control normotensive subjects were studied. The M235T AGT and the I/D ACE gene polymorphisms were determined by polymerase chain reaction (PCR). Family and personal history of cardiovascular disease, lipoprotein profile, microalbuminuria and left ventricular hypertrophy (LVH) by Sokolow index were analyzed in hypertensive patients. RESULTS: The distribution of the different polymorphisms was similar among hypertensive and normotensive subjects. There was not any relationship among AGT nor ACE genotypes and target organ damage. The II ACE genotype was associated with higher lipoprotein (a) (Lp[a]) levels and greater cerebrovascular disease family history and the MT AGT genotype with lower total cholesterol (TC) and triglycerides (TG) levels. CONCLUSIONS: In our study there was not any relationship between arterial hypertension and M235T AGT or I/D ACE gene polymorphisms. None specific genotype was associated with worse cardiovascular risk profile. The II ACE genotype was a predictor of cerebrovascular disease risk through higher levels of Lp(a) and the MT AGT genotype was associated with a better lipid profile. PMID- 10480139 TI - [The evolution of AIDS mortality and survival in Barcelona (1981-1997)]. AB - BACKGROUND: To analyze the evolution of AIDS mortality and survival in Barcelona, Spain. PATIENTS AND METHODS: AIDS-cases registered in Barcelona (Spain) between 1981-1997. Study of mortality and survival probability. RESULTS: Mortality decreased by 55% between 1996-1997. Survival probability at 547 days was of 80.3% in 1997 compared with 48.6% before 1988. Being older than 36 years old, IVDU, having other AIDS-defining disease than tuberculosis, and year of diagnostic before 1997 were associated with poor survival. CONCLUSIONS: AIDS mortality in Barcelona has enormously decreased. The best survival in 1997 is probably attributable to the new antiretroviral therapies. PMID- 10480140 TI - [AIDS: more and better life?]. PMID- 10480141 TI - [Sensitivity-based medicine]. PMID- 10480142 TI - [Severe liver failure in erythropoietic protoporphyria. A report of a case treated by liver transplantation]. AB - Severe liver failure is a rare complication of erythropoietic protoporphyria (PEP), which is associated with a high rate mortality. Until now, 31 patients with this hepatic complication had underwent a liver transplantation, with a high rate of survival, but their long-term outcome is not well established. We report the first case in Spain of PEP in 59-year-old, whose acute liver failure was treated with liver transplantation, without postoperative complications. The patient is in good clinical condition 30 months later. Nevertheless during the first eleven months of follow-up the plasma levels of protoporphyrin remained elevated, which was accompanied of biochemical and histological evidence of relapse of the metabolic disease in the graft. Cases such as this stress the usefulness of liver transplantation, but also the need of more efficient measures to decrease the protoporphyrin levels before and after the transplant, in order to prevent hepatic and extrahepatic complications in these patients. PMID- 10480143 TI - [Of privatizations and "analyzables"]. PMID- 10480144 TI - [The usefulness of CK-MB isoforms in heart disease]. PMID- 10480145 TI - [Bilateral lung images and respiratory insufficiency in an 86-year-old man with polyarteritis nodosa]. PMID- 10480146 TI - [Pregnancy in an acromegalic patient treated with lanreotide and bromocryptin]. PMID- 10480147 TI - [The Gordon syndrome: the first reported case in Spain]. PMID- 10480148 TI - [Renal involvement in patients with chronic inflammatory intestinal disease treated with mesalazine. How to prevent its nephrotoxicity?]. PMID- 10480149 TI - [Xenotransplantation: from the myth to the possible]. PMID- 10480150 TI - [Can man live with a pig kidney?]. AB - The transplantation of organs from one species to another introduces a question of compatibility not seen in allotransplantation, the ability of a kidney to perform its physiological function in the new host environment. It has been assumed that an allotransplanted organ will function normally if is not rejected; ample experience supports this assumption. This luxury will not exist in the field of xenotransplantation, where the issues of comparative physiology will assume great importance. From many standpoints, the pig kidney seems an ideal donor for xenotransplantation. They are of similar size and have remarkably similar internal anatomy. Even if the immunological problems could be overcome, there is almost no direct experimental evidence to answer the question of whether or not a pig kidney can function in a human body. PMID- 10480151 TI - [Regulation of intracellular cAMP content in kidney tubules: role of adenylyl cyclases inhibitable by calcium]. AB - Adenylyl cyclase (AC) isoforms 5 and 6 can be inhibited by submicromolar concentrations of Ca2+. Quantitative RT-PCR allowed to study the corresponding messengers (mRNA) along the rat nephron. The results demonstrate a significant expression of AC 6 mRNA all along the nephron and of AC 5 mRNA in the glomerulus and the collecting tubule located in the cortex and the outer medulla. Regulation of cAMP synthesis and of intracellular cAMP content in defined renal cell types established the functional expression of AC 5 and AC 6. In particular, adenylyl cyclase activity is strongly stimulated by hormones and can be inhibited by several factors which either increase intracellular Ca2+ concentration or are coupled to G alpha 1. In each renal cell studied, the expression of 5 and 6 isoforms allow to integrate specific, multiple and independent inhibitory pathways which contribute to decrease intracellular cAMP content. PMID- 10480152 TI - [Antihypertensive treatment in type 1 diabetes and diabetic nephropathy]. AB - In diabetic patient, hypertension is a major factor contributing to both cardiovascular morbidity-mortality and progression toward renal impairment. This review analyzes studies from the literature regarding the benefits of antihypertensive treatment at every stage of type 1 diabetes and diabetic nephropathy. PMID- 10480153 TI - [Role of the time of administration of calcium carbonate (before or during mealtime) in the control of hyperphosphatemia in patients on maintenance hemodialysis]. AB - A study has claimed that at equal elemental calcium dose, CaCO3 was not less but equally efficient in controlling predialysis hyperphosphatemia as calcium acetate provided both calcium salts were given five minutes before the meals instead of during the meals because the higher acidity of a fasting gastric juice would allow a better dissociation of CaCO3. To examine the point that CaCO3 is more efficient if it is taken five minutes before compared to during the meal, we performed a two month randomised cross-over trial in twelve reliable and stable patients maintained on chronic hemodialysis while their treatment and diet remained constant. Comparison of the plasma concentrations measured during the two modes of administration showed no significant difference in creatinine, urea, bicarbonate, intact-PTH. Mean (+/- SD) plasma PO4 was significantly higher (1.93 +/- 0.50 versus 1.72 +/- 0.40 mmol/l; p = 0.02) whereas corrected plasma Ca was significantly lower (2.30 +/- 0.15 versus 2.38 +/- 0.17 mmol/l; p = 0.01) when CaCO3 was given before the meals than during the meals. CONCLUSIONS: a) administration of CaCO3 before the meal decreases its efficiency in controlling hyperphosphatemia since plasma PO4 was actually slightly higher with this timing of administration; b) administration of CaCO3 before the meal is associated with significantly lower plasma corrected calcium suggesting a smaller absorption of calcium which may be an advantage but only in hypercalcemic patients; c) there is no reason other than the prevention of its hypercalcemic effect to recommend the administration of CaCO3 just before the meals than during the meals. PMID- 10480154 TI - [Inhibition of gastric secretion by omeprazole and efficacy of calcium carbonate in the control of hyperphosphatemia in patients on maintenance hemodialysis]. AB - Apparent contradictions exist in the literature regarding the effect of gastric secretion inhibition on phosphatemia. In healthy adults, omeprazole has been reported to prevent the increase of plasma phosphate or of phosphaturia observed after an acute phosphate load suggesting an inhibition of phosphate absorption. In patients on chronic hemodialysis their hyperphosphatemia is associated to gastric hypersecretion but the inhibition of this latter by ranitidine in patients taking CaCO3 has been reported to increase their plasma phosphate. Because of this contradiction, we have made an open cross-over study in 16 stable and compliant patients in chronic hemodialysis taking a mean daily dose of 9.4 +/ 4 g of CaCO3 and compared their predialysis plasma concentration of phosphate, calcium, protides, bicarbonates, intact PTH, urea and creatine for two successive periods of two months without or with 20 mg of omeprazole. Plasma phosphate did not increased significantly with omeprazole (from 1.80 +/- 0.38 to 1.89 +/- 0.42 mmol/l) whereas plasma corrected calcium significantly decreased from 2.41 +/- 0.18 to 2.36 +/- 0.16 mmol/l (p = 0.04) and plasma bicarbonate decreased significantly from 26.7 +/- 3.5 to 25.7 +/- 3.1 mmol/l (p < 0.05). No significant change was observed in plasma creatine and urea suggesting stability of dialysis efficiency and of protein and therefore phosphate intake. CONCLUSION: These data do not support that CaCO3 efficiency as phosphate binder is decreased with inhibition of the gastric secretion by omeprazole. PMID- 10480155 TI - Evolutive aortic stenosis in hemodialysis patients: analysis of risk factors. AB - Cardiovascular diseases represent the major cause of death in hemodialysis patients. However, little information is available about the repercussion of uremia on cardiac valves. We retrospectively investigated the incidence rate of aortic stenosis (AS), from 1991 to 1996, in 110 hemodialysis patients followed by Doppler-echocardiography. Progressive AS was diagnosed in 16 patients who had a decrease in their indexed aortic valve area from 1.24 +/- 0.09 to 0.66 +/- 0.21 cm2/m2 of BSA in 16.8 +/- 1.9 months. The mean incidence of AS per year was of 3.3%, ranging from 1.5 to 8.0%. Eight patients died in less than 3 years after the diagnosis of AS with a mean survival time of 23.0 +/- 9.5 months. Survival curves using Kaplan-Meier estimates showed a statistically significant decrease in the survival rate of patients with AS compared with patients without valvulopathy (p < 0.001). They were older than patients with normal valve, 68.6 +/- 11.1 versus 56.7 +/- 16.0 years, respectively. Men were 4 times more affected than women and showed a significantly more rapid progression to AS than women. The calcium-phosphorus product was higher in AS patients, 5.43 +/- 0.98 than in patients without AS, 3.95 +/- 0.50 mM. It was mainly due to hyperphosphatemia without hypercalcemia and the hyperphosphatemia was associated with biological signs of hypoparathyroidism or adynamic bone disease in 62% of the cases. Plasma vitamin D3 was also higher in patients with AS, 20.5 +/- 13.5 ng/ml than in those with normal valves, 9.6 +/- 6.3 ng/ml. Logistic regression showed that age, vitamin D3 and hyperphosphatemia correctly predicted 56% of the AS cases. In conclusion, AS is frequent and of poor outcome in hemodialysis patients. Age, relatively high plasma vitamin D3 levels, and hyperphosphatemia, mostly due to hypoparathyroidism, must be considered as risk factors. PMID- 10480156 TI - [Ligand recognition and activation mechanism of histamine H1 receptor]. AB - Five amino acid residues of the human histamine H1 receptor that participate in histamine binding were identified using mutant H1 receptors by site-directed mutagenesis and 3D computer modeling of the receptor. The computer modeling provided two conformations of the receptor, non-active and active forms, and it was hypothesized that the binding of histamine to the receptor leads to the active state through the rewinding of the alpha-helix of TM-V. The binding sites of histamine were divided into two groups, one determined the ligand affinity and the other directly participated in the conformational change of the receptor, and the H1 antagonists bound only to the former. The mode of binding of histamine to the H1 receptor was different from those of catecholamine to the beta-receptor and of histamine to the H2 receptor. PMID- 10480157 TI - [Pharmacological effects of the gastroprokinetic agent mosapride citrate]. AB - Mosapride citrate (mosapride) is a novel gastroprokinetic agent that enhances the gastrointestinal motility by stimulating the 5-hydroxytryptamine4 (5-HT4) receptor. Mosapride dose-dependently enhanced the gastric emptying of a liquid or solid meal in rats with a potency equal to that of cisapride and more potent than that of metoclopramide. In rats, mosapride improved the gastric emptying delayed by gastroduodenal surgical intervention. In the conscious dogs with force transducers implanted chronically, mosapride stimulated antral and duodenal motility with a potency equal to those of cisapride. In isolated guinea-pig ileal longitudinal muscle preparations, mosapride enhanced the electrically stimulated contractions, and the enhancing effect of mosapride was antagonized by a high dose of tropisetron, a 5-HT4-receptor antagonist. In addition, mosapride inhibited [3H]-GR-113808 binding to 5-HT4 receptor sites of guinea-pig ileum and striatum. Mosapride had no affinity for dopamine D2 receptor, whereas metoclopramide and cisapride had a high affinity for dopamine D2 receptor. In isolated guinea-pig papillary muscles, mosapride did not prolong the duration of action potentials, whereas cisapride concentration-relatedly prolonged it. In conclusion, mosapride is a selective and potent 5-HT4 receptor agonist and improves gastrointestinal symptoms in patients with non-ulcer dyspepsia without causing the extrapyraminal syndrome associated with dopamine-D2-receptor blockage and adverse cardiovascular effects such as torsadoes de points. PMID- 10480158 TI - [Molecular dissection by gene targeting in the chicken B cell line DT40]. AB - The gene-targeting experiment is a powerful tool for clarifying the functions of each gene product. For instance, establishment of a gene-knockout mouse, undoubtedly, contributes to our understanding of the functions of each gene in vivo. However, assuming that the gene product is expressed in various lineage cells, the phenotype in this knockout mouse exhibits a mixture of defects in various lineage cells. Thus, to compensate for this weak point, cell-based gene knock experiments are required. Since the DT40 chicken B cell line shows a high frequency of homologous recombination, this line can be used for rapid dissection of each gene product at the cellular level. PMID- 10480160 TI - [Regulation of bacteriophage lambda and lambda plasmid DNA replication]. PMID- 10480159 TI - [Effects of benidipine hydrochloride (Coniel) on blood pressure, heart rate and plasma norepinephrine concentration in spontaneously hypertensive rats]. AB - We investigated the effects of benidipine hydrochloride (benidipine, Coniel) on blood pressure, heart rate and plasma norepinephrine (NE) concentration in spontaneously hypertensive rats and compared them with those of other calcium channel blockers. Benidipine (2 mg/kg, p.o.) was compared with the equihypotensive doses of nifedipine (5 mg/kg), cilnidipine (6 mg/kg) and amlodipine (3 mg/kg). All the 4 calcium channel blockers exhibited significant antihypertensive effects. Nifedipine and cilinidipine significantly increased heart rate, as compared with that in the control group, whereas benidipine or amlodipine did not significantly affect it. The area under the curves for hypotensive effect and tachycardic effect for 10 hr after the drug administration were compared among the 4 compounds. As a result, the tachycardic effect of benidipine was significantly lower than those of nifedipine, cilnidipine and amlodipine, while the hypotensive effects were similar among the 4 compounds. Nifedipine and amlodipine, significantly increased plasma NE concentration, cilnidipine tended to increase it. In contrast, benidipine did not significantly affect plasma NE concentration. These results suggest that the effects of benidipine on plasma NE concentration and heart rate are less prominent than those of the other calcium channel blockers. PMID- 10480161 TI - [Regulation of mammalian muscle myosin gene expression]. PMID- 10480162 TI - [Signal transduction of nerve growth factor (NGF) and its TrkA and p75NTR receptors]. PMID- 10480163 TI - [Intracellular proteolysis]. PMID- 10480164 TI - [Urokinase plasminogen activation system and its role in cancer progression]. PMID- 10480165 TI - [The role of reactive oxygen species in mutagenesis and carcinogenesis processes]. PMID- 10480166 TI - [Adaptation of thylakoid membranes to environmental light conditions]. PMID- 10480167 TI - [Have you heard of Barbara McClintock?]. PMID- 10480168 TI - [Prevalence of antiphospholipid antibodies in Horton's disease and in polymyalgia rheumatica]. AB - PURPOSE: Antiphospholipid antibodies (APL) are frequently observed in the course of giant cell arteritis and polymyalgia rheumatica. However, their role and relationships with potential ischemic events are still debated. METHODS: To determine the prevalence of APL in relation with ischemic events, 62 patients with giant cell arteritis and/or polymyalgia rheumatica were retrospectively studied. RESULTS: Before corticosteroid treatment 41% of the 51 patients with giant cell arteritis and 64% of the 11 patients with isolated polymyalgic rheumatica had high IgG ACL levels, with a frequency significantly higher than that (15.6%) of the control group which was composed of healthy elderly. IgM ACL were found in only two patients with giant cell arteritis. There was no correlation between the occurrence of an ischemic event (22 ischemic events in the 51 patients with giant cell arteritis) and the presence of ACL, even though the latter were more frequently observed in the giant cell arteritis group (52% versus 41% in non ischemic patients). The prevalence of ACL increased, reaching 59% if lupus anticoagulant was associated, but the difference was not significant. ACL disappeared soon after corticosteroid therapy had been initiated. CONCLUSION: Though ACL are frequently seen in giant cell arteritis and/or polymyalgia rheumatica, they are not related to ischemic events and disappear rapidly after corticosteroid treatment. PMID- 10480170 TI - [Mechanisms of paraneoplastic neurologic syndromes]. AB - INTRODUCTION: Paraneoplastic neurological syndromes (PNS) refer to a set of neurological disorders associated with neuronal degeneration in some patients with systemic cancers. These disorders are not related to the tumor mass or metastasis. CURRENT KNOWLEDGE AND KEY POINTS: Evidence argues for an autoimmune reaction against tumor cells which expresses antigens normally present in neurons. A high percentage of patients with PNS harbors high titers of anti neuronal autoantibodies in their serum and cerebrospinal fluid. In addition to their clinical interest in diagnosis and pathophysiology, these autoantibodies provide a unique opportunity to identify genes encoding previously undiscovered neuronal proteins which are also expressed by tumor cells. These "onconeural" antigens have been classified in four groups: neuromuscular junction proteins, nerve terminal/vesicle-associated proteins, neuronal RNA binding proteins, or neuronal signal transduction proteins. FUTURE PROSPECTS AND PROJECTS: All of these proteins would play a major role in the neuronal maturation and homeostasis, and for some of them in cellular proliferation. Better understanding of the exact role of these proteins would in turn permit better understanding of the mechanisms of neuronal degeneration and tumor cell proliferation in PNS. PMID- 10480169 TI - [Requirements for the successful implementation of unity of care: study of a computer-assisted dispensation of pharmaceuticals]. AB - PURPOSE: Even though computerized workstations bring undisputed benefits in nursing units, introducing them is still hard when most of the staff members have to share the workstation. We took advantage of the implementation of the drug prescription software SAUPHIX in a nephrology department to better define the encountered difficulties. The workstation described in this paper is shared by physicians who enter their prescriptions (proprietary names, doses, routes of administration), nurses who use dosage schedules for drug administration, and the chemist who has authority to control prescription orders. METHODS: Six months after the implementation of the workstation, physicians and nurses had to fill out an anonymous questionnaire aimed at assessing each function of the software. RESULTS: Prescriptions proved to be more accurate and legible, while management of drugs was more precise. However, interns complained that entering data was time consuming. Furthermore, they raised objections to control of prescription orders. Nurses criticized dosage schedules, the primary reason being that they had to change their practice. The convenience of notebooks was questioned by both physicians and nurses who would have preferred a greater number of desktop computers at their disposition. CONCLUSION: The implementation of a computerized workstation requires information, diplomacy and negotiations to obtain real implication of the staff. Tasks and schedules must be specified for everybody. The system has to be carefully customized, according to the requirement of the unit. Computers must be properly chosen and allocated in sufficient number. Finally, appropriate preparation, staff training and follow-up of the computerized system are essential. PMID- 10480171 TI - [Contribution of clinical epidemiology to the management of HIV infections]. AB - INTRODUCTION: The objective of this publication is to make clinicians more aware of clinical epidemiology through our experience in the field of human immunodefiency virus (HIV) infection. CURRENT KNOWLEDGE AND KEY POINTS: Clinical epidemiology is aimed at studying diagnostic, therapeutic and prognostic aspects of diseases, and their consequences, with the objective of improving both scientific knowledge and patient's management. Examples, including longitudinal studies and clinical trials, illustrate the value of clinical epidemiology. FUTURE PROSPECTS AND PROJECTS: Beyond HIV infection, the importance of both multidisciplinary collaboration and education of clinicians in regard to biostatistical and epidemiological methods is emphasized. PMID- 10480172 TI - [Danger of anti-erythrocyte antibodies during pregnancy]. AB - INTRODUCTION: Red cell autoantibodies are very rarely observed in pregnant women. We report one case related to the presence of a lupus anticoagulant. EXEGESIS: A 32-year-old woman had a history of two intrauterine deaths that had occurred at weeks 32 and 33, respectively. Due to the presence of a lupus anticoagulant, prednisone (20 mg/day) and subcutaneous calcium heparin were administered during her third pregnancy. The direct antiglobulin test remained positive throughout the pregnancy; however, hemolysis could not be detected. Caesarean section was done in emergency at week 29, due to fetal bradycardia related to severe fetal hemolysis. CONCLUSION: When autoimmune hemolysis occurs during pregnancy, risks for the fetus increase with the severity of hemolysis in the mother. It is thought that child mortality reflects the existence of anemia, rather than the existence of hemolysis. Our observation suggests that severe fetal hemolysis may occur, despite the absence of hemolysis in the mother. PMID- 10480173 TI - [Bronchopulmonary cancer, antiphospholipid syndrome and coagulation disorders]. AB - INTRODUCTION: The antiphospholipid syndrome includes recurrent thrombotic manifestations related to antiphospholipid antibodies. Adrenal insufficiency is a rare complication of the antiphospholipid syndrome. EXEGESIS: We report a case of acute adrenal insufficiency secondary to bilateral adrenal hemorrhage in a 45 year-old man. The finding of antiphospholipid antibodies and 6 months later of a polymetastatic bronchopulmonary cancer led us to diagnose a paraneoplasic antiphospholipid syndrome. CONCLUSION: We discuss the role of coagulation disorders in the pathogenesis of tumor growth and rapid metastatic spread. Assessment of the high risk for thrombosis may be of prognostic and therapeutic value in patients with evolutive bronchopulmonary cancer. Early anticoagulation treatment in association with classical treatment of cancer may contribute to prevent malignant process from extending and avoid metastatic spread. PMID- 10480174 TI - [Group B streptococcal meningitis revealing an inner ear congenital malformation. A case report in an adult]. AB - INTRODUCTION: Group B streptococcal meningitis is unusual in adults. It occurs in patients with a chronic disease. EXEGESIS: We report a case of group B streptococcal meningitis that occurred in a 32-year-old woman, uncovering inner ear congenital malformation associated with cerebrospinal fluid fistula. There was no recurrent episode of meningitis during a 2-year period following surgical correction of the malformation. CONCLUSION: Clinical signs suggesting cerebrospinal fluid fistula should be considered in any adult patient with severe group B streptococcal infection. PMID- 10480175 TI - [Primary sarcoma of the pulmonary artery. Case report]. AB - INTRODUCTION: Sarcoma of the common pulmonary artery is a rare malignant tumor that can mimic pulmonary embolism. EXEGESIS: We report a case of a pulmonary artery sarcoma that occurred in a 60-year-old woman and had an unusual (4-year duration) evolution. CONCLUSION: Early diagnosis with adequate surgical procedures (complete resection) leads to better prognosis. PMID- 10480176 TI - [Genetics of Parkinson disease]. AB - INTRODUCTION: What is the role of genetic factors in the pathophysiology of idiopathic Parkinson's disease, one of the most frequent neurodegenerative disorders? In the past two years, identification of two genes and localization of a third one have supported the hypothesis that genetics factors are involved in idiopathic Parkinson's disease. We present arguments that support such hypothesis, and describe recent advances in genetic studies of idiopathic Parkinson's disease. CURRENT KNOWLEDGE AND KEY POINTS: The first gene identified on chromosome 4 encodes alpha-synuclein. It causes a rare form of autosomal dominant Parkinson's disease. A locus on the short arm of chromosome 2 was recently identified in families with autosomal dominant Parkinson's disease. More recently, the gene encoding Parkin (located on chromosome 6) has been described. It already appears to be an important locus for juvenile parkinsonism with autosomal recessive transmission. CONCLUSION: We now have to understand how mutations in these genes lead to selective degeneration of dopaminergic neurons, and to determine whether or not they participate in the genetic susceptibility of idiopathic Parkinson's disease. PMID- 10480177 TI - [Diffuse pulmonary calcification]. PMID- 10480178 TI - [Peripheral T-cell lymphoma 9 months after hemophagocytic syndrome with a favorable outcome after splenectomy]. PMID- 10480179 TI - [Bupivacaine-induced meningitis]. PMID- 10480180 TI - [Hypokalemic periodic paralysis revealing hyperthyroidism]. PMID- 10480181 TI - [Osteochondromatosis associated with aseptic osteonecrosis during systemic lupus]. PMID- 10480182 TI - [Problems of bone mineralization secondary to ureter sigmoid diversion]. PMID- 10480183 TI - [Antiviral treatment of hepatitis C virus infection]. AB - Hepatitis C virus infection is common and almost always chronic and can lead to cirrhosis and hepatocellular cancer. The primary goal of the treatment is virus eradication and the secondary is to reduce inflammation and liver cell damage. Interferon is the only effective therapy but disappearance of the virus is sustained in only 10 to 15%. The factors most closely associated with a response to treatment are absence of cirrhosis, low serum hepatitis C virus RNA level and genotype other than type 1. Recent studies have suggested that interferon treatment may reduce the subsequent risk of hepatocellular carcinoma in responders. The combination of interferon and oral ribavirin therapy increase the sustained response rate to about 40% in initial treatment and 50% for the treatment of relapse. The other therapeutic combination are less well documented. New agents such as hepatitis C virus-specific anti-protease may be available in the next future and treatment is evolving toward multiple-drug regimens. PMID- 10480184 TI - [New nonsteroidal anti-inflammatory agents: nitric oxide donors and selective cyclooxygenase-2 inhibitors]. AB - The use of non steroidal anti-inflammatory drugs as analgesic or anti inflammatory agents is primarily limited by their toxicity to the gastrointestinal tract. Two strategies have been developed recently in order to improve the safety of these drugs. The first approach is the linking of a nitric oxide-releasing moiety to the available compounds. The rationale is that nitric oxide may prevent non steroidal anti-inflammatory drugs-induced ulcerations by preventing mucosal ischemia. The second approach is based on the discovery of two isoforms (COX-1 and COX-2) of the cyclo-oxygenase enzyme. It was hypothesized that the constitutively expressed COX-1 isoenzyme leads to the synthesis of prostaglandins with homeostatic functions whereas COX-2 is merely responsible for the production of prostaglandins mediating pain, fever and inflammation. Accordingly, selective COX-2 inhibitors have been developed. Clinical trials indicate that these compounds are roughly as effective as the available non steroidal anti-inflammatory agents without causing acute gastrointestinal damage. There is some evidence that both COX-1 and COX-2 isoforms are involved in the production of prostaglandins associated with inflammation and homeostatic functions. Finally, the true benefit/risk ratio of these new non steroidal anti inflammatory drugs remains to be assessed. PMID- 10480185 TI - [New therapeutic strategies in multiple sclerosis]. AB - Until a few years ago, the focus of MS therapy was mainly on the symptomatic approach. Several rigorous clinical trials concerning mainly relapsing-remitting forms were conducted recently. As a result of these, therapeutical views have changed. Elevated doses of methylprednisolone, used during a short period have significantly improved the handicap; but impact over a longer time is unknown. It has now been demonstrated that beta interferons reduce the frequency of relapses, modify the handicap favourably and improvements are clearly shown on MRIs. Therefore MRI has become an invaluable tool for the evaluation of the efficacy of new drugs. Recently, there have been positive results made in progressive MS. Two other drugs, Copolymer and Immunoglobulins have also shown encouraging results but other studies are still necessary. The role of some immunosuppressive agents, but mainly the role of mitoxantrone are now better understood. These results are encouraging but they have also raised a lot of questions such as: how can these drugs be used in other forms of MS; what is their long-term impact on the disease; what is the mechanism of their action; what is the etiopathogeny of MS? Clinical trials are being conducted to answer these questions and to study the usefulness of combined therapies. Even though these results have been positive, other therapies which focus on spasticity, fatigue, sphincter, dysfunction and psychosocial problems must not be neglected. PMID- 10480186 TI - [Present status in the treatment of type 2 diabetes mellitus. Insulin-secreting agents]. AB - The functional defect of the pancreatic beta cell represents one of the main therapeutic targets in type 2 diabetes mellitus. Among the currently available oral antidiabetic drugs, only hypoglycaemic sulfonylureas exhibit beta cell stimulating properties. However, their use has some limits, particularly those related to the risk of hypoglycaemia and the frequent secondary therapeutic failure. These drugs have largely contributed to the knowledge of the mechanisms of insulin secretion. Besides some galenic modifications of existing sulfonylureas and the development of new drugs of this family with original properties, like glimepiride, the research is essentially focused on drugs derived from the non sulfonylurea moiety of some sulfonylureas, particularly the meglitinide family, which will probably be available for the clinician in the near future. These drugs act however grossly by the same mechanism than sulfonylureas, even if their binding site on a protein coupled with the ATP sensitive K channel appears different. Among the other possible approaches suggested by the theoretical data concerning the mechanisms of insulin secretion, GLP-1 derivatives probably represent good candidates, if stable analogues are developed. PMID- 10480187 TI - [Primary nutritional and drug prevention of atherosclerosis]. AB - Atherosclerosis results from a multifactorial process. For that reason, primary prevention of cardiovascular diseases requires several measures that should exhibit antiatherogenic, antithrombotic, antioxidative and antihypertensive properties. Two types of therapeutic strategies can be individualized. Nutritional interventions are the first line measures for population wide primary prevention in subjects at low risk of developing atheroscleric diseases. In high risk individuals who have either one major risk factor (LDL cholesterol above 190 mg/dL) or several additional risk factors, dietary measures must be frequently combined with drug treatments. However, the cost-effectiveness ratio should be weighed prior to any treatment especially with statins. Dietary measures consist to reduce caloric intakes in overweight subjects, or more generally to adopt eating patterns which correspond to the following recommendations: (i) proteins = 15% of total calories; (ii) saturated and polyunsatured fats = 10% each; (iii) carbohydrates plus monounsaturated fats = 65%. Such dietary instructions permit individual dietary changes, just by adjusting the carbohydrates/monounsaturates balance, the sum of both nutrients remaining in all cases equal to two-thirds to total calories. Drug treatments are only indicated after secondary dietary failure. They are generally based on the use of statins since the efficacy of these treatments has been clearly established by such preventive trials as the WOSCOP Study, provided that the therapeutic interventions result in significant reduction of LDL cholesterol levels over several years. Antiplatelet agents (aspirin), antihypertensive therapies, antioxidant supplementations with vitamins E or C are also recommended in some individuals for completing the beneficial effects of the above mentioned measures. PMID- 10480188 TI - [Current status of the treatment of type 2 diabetes mellitus. The revival of insulin-resistance drugs]. AB - The relief of insulin resistance is one of the two therapeutic targets of the treatment of type 2 diabetes. Insulin-sensitizers are therefore complemental with other oral diabetic drugs. The treatment of insulin resistance was for a long time limited to dietary and exercise programmes, a biguanide, metformine, and benfluorex, a phenylethylamine derivative; the mechanisms of action of both drugs are now better understood and their indications more precisely targeted. A new therapeutic class, the thiazolidinediones (troglitazone, rosiglitazone, pioglitazone) has recently completed the family of insulin-sensitizing agents. These drugs, which should be soon available in France, act by a different way than metformin, which has been recently identified as the peroxisomes proliferator-activated receptor. The role of antilipolytic agents, which might increase glucose uptake by reducing free fatty acid production and oxidation is under evaluation, as well as the potential benefit of orlistat, an inhibitor of lipid digestion which has been proved effective, in addition to hypocaloric diet, in the management of obese patients. PMID- 10480189 TI - [Current status of the treatment of type 2 diabetes mellitus. Alpha-glucosidase inhibitors]. AB - The development of antidiabetic drugs with complementary mechanisms of action appears more and more necessary in order to achieve durable glycaemic control in type 2 diabetes. By inhibiting in a reversible way the hydrolysis of disaccharides and the ultimate steps of the digestion of dietary polysaccharides, alpha-glucosidase inhibitors reduce postprandial blood glucose raise in diabetics. This therapeutic class, limited in Europe until recently to acarbose, has been enlarged with the marketing of miglitol, whose pharmacokinetic properties might lead to better long term tolerance. The improvement of glycaemic control obtained with alpha-glucosidase inhibitors is now better evaluated and appears similar whatever the combinations with other antidiabetic drugs, including insulin. The role of alpha-glucosidase inhibitors in the overall therapeutic strategy of type 2 diabetes and their benefit on the evolution of long term complications remains to be clarified. PMID- 10480190 TI - [Methotrexate in dermatology: pharmacology, indications, applications and prudent use]. AB - High dosage methotrexate is currently used in the treatment of malignancies. When used at low- or moderate doses, methotrexate has antiproliferative and antiinflammatory effects and is a useful drug in skin diseases. The aim of this review is to describe pharmacology, indications, adverse effects and practical use in Dermatology. Pharmacodynamics of methotrexate is especially related to antifolic activity. Methotrexate is officially approved in the treatment of severe psoriasis, but many other proliferative or inflammatory diseases with cutaneous manifestations may benefit from this drug, usually in association with corticosteroids. The use of methotrexate needs some precautions and a precise follow-up to minimise the risk of severe adverse effects. However, efficacy of methotrexate was reported in open and retrospective small size studies. Prospective and comparative trials are required to confirm the indications, advantages and tolerance of methotrexate in dermatology. PMID- 10480191 TI - [A comparative long-term trial of sodium cerivastatin, a new HMG-CoA reductase inhibitor, in patients presenting with primary hypercholesterolemia]. AB - Cerivastatin sodium a synthetic and pure enantiomeric 3-hydroxy-3-methylglutaril coenzyme A (HMG-CoA) reductase inhibitor, is considered effective in the treatment of mild-to-moderate primary hypercholesterolemia (total cholesterol < or = 220-259 mg/dL) at a daily dose of 0.15 mg. We compared the efficacy and tolerability of a dosage of 0.3 mg/d with those of a dosage of 0.15 mg/d in patients with severe primary hypercholesterolemia (serum total cholesterol > or = 260 mg/dL). After a minimum of 4 week's lead-in with placebo, 73 patients with severe primary hypercholesterolemia were randomly assigned to receive either 0.15 or 0.3 mg of cerivastatin sodium once daily after the evening meal for 12 weeks. In 58 patients, the same drug was continued at a flexible dosage for an additional 36 weeks or longer to assess the long-term efficacy and tolerability of cerivastatin sodium. During the 12-week treatment period, serum total cholesterol levels decreased significantly from baseline in both dosage groups, but the percentage reduction was significantly greater in the 0.3-mg group (range, 24.4% to 25.6%) than in the 0.15-mg group (range, 19.4% to 21.6%). The percentage reduction in levels of low-density lipoprotein cholesterol, triglycerides, and apolipoprotein B and the percentage increase in levels of high density lipoprotein cholesterol were significantly greater in the 0.3-mg group than in the 0.15-mg group. When the results for the 0.3- and 0.15-mg groups were combined, the percentage of change in serum lipid levels at 48 weeks remained as stable as at 12 weeks. No serious adverse reactions were observed. We concluded that the higher dose of cerivastatin sodium was more effective than the lower dose, with comparable tolerability, in the treatment of patients with severe primary hypercholesterolemia. PMID- 10480192 TI - [The treatment of diabetes insipidus in 1999]. PMID- 10480193 TI - Improving outcomes for recent-onset psychoses: disentangling hope, speculation and evidence. PMID- 10480194 TI - Cognitive deficits and cognitive training in schizophrenic patients: a review. AB - OBJECTIVE: The aim of this paper is to review the current status of knowledge of cognitive deficits and remediation in patients with schizophrenia. METHOD: Relevant reports were identified by a literature survey. In addition, some outstanding researchers in these areas were asked to add to the identified list relevant literature that was not included. RESULTS: Our review focuses on the cognitive deficits observed in the areas of attention, memory and executive functions. We attempt to classify dysfunctions as vulnerability- or symptom linked factors, and we discuss the methodological question of a general performance deficit vs. a differential deficit. Furthermore, we briefly delineate how antipsychotics affect cognitive functions. Finally, controlled studies of cognitive training are discussed in more detail. CONCLUSION: The most outstanding cognitive dysfunctions in patients with schizophrenia can be related to the areas of attention, memory and executive functions. Interest in cognitive remediation has to some extent been rekindled in the 1990s. However, few studies on the effects of cognitive training programs have been conducted. PMID- 10480195 TI - Duration of untreated psychosis and 12-month outcome in first-episode psychosis: the impact of treatment approach. AB - OBJECTIVE: Early intervention research is examining whether reducing the duration of untreated psychosis (DUP) leads to improved outcome from first-episode psychosis. Another key influence may be the quality of treatment after initiation of care. This study examined the effect of phase-specific treatment on 12-month outcome for different categories of DUP. METHOD: A total of 250 first-episode psychosis cases were followed up 12 months after stabilization. The sample consisted of two historically sequential cohorts treated in the same region within different service models, one of which was more intensive and phase specific. Outcome was compared according to four predefined categories of DUP. RESULTS: Only patients with a mid-range DUP of 1-6 months who were treated within the phase-specific model experienced significantly better outcomes than patients treated within the previous model. CONCLUSION: These data suggest that there may be a limited window of opportunity in which to influence outcome. However, the complexity of this relationship is emphasized. PMID- 10480196 TI - Depression, negative symptoms, social stagnation and social decline in the early course of schizophrenia. AB - OBJECTIVE: The aim of this study was to investigate when social consequences in schizophrenia emerge, and what conditions give rise to the social disadvantage evident in people suffering from schizophrenia. METHOD: Early course in schizophrenia was studied in a population-based sample of 232 first illness episode cases retrospectively from onset to first admission, and in a representative subsample of 115 patients prospectively at six cross-sections over a period of 5 years. Data on non-specific and negative symptomatology and social development was compared with data from an age- and sex-matched control group drawn from the normal population. RESULTS: In total, 73% of the patients showed a prodromal phase of several years. First signs were depressive and negative symptoms. In 57% of cases social disability emerged 2 to 4 years before first admission. Social consequences depended on the level of social development at onset. An early onset involved social stagnation, and a late onset was associated with social decline. Men's poorer social outcome was determined by their lower level of social development at onset and socially adverse illness behaviour. The 5-year symptom-related course showed no gender difference. At 81% the lifetime prevalence of depressive mood until first admission was several times higher in schizophrenics than in healthy controls. Early depression predicted a lower subsequent score for affective flattening. Suicide indicators were predicted by lack of self-confidence and feelings of guilt early in the illness. CONCLUSION: Taking into account a prodromal phase of several years on average before first hospital admission, early detection, case identification and intervention are urgently needed. The intervention must be targeted at syndromes such as early depression, negative symptoms and certain forms of cognitive and social impairment. PMID- 10480197 TI - Neurological abnormalities in schizophrenic patients: a prospective follow-up study 5 years after first admission. AB - OBJECTIVE: The aim of this study was to examine the temporal stability of neurological abnormalities in first-episode schizophrenic patients, and to clarify the relationships between such abnormalities and psychopathology. METHOD: A total of 18 schizophrenic patients, 11 non-schizophrenic patients and 10 healthy volunteers were examined neurologically at first admission and 5 years later. RESULTS: A significant increase in the number of neurological abnormalities was seen in schizophrenic patients with genetic predisposition and in patients with a non-remitting course of disease. Birth complications had an impact on the occurrence of neurological abnormalities at first admission. CONCLUSION: The association between a deteriorating course of disease and neurological impairment supports the theory that any possible impairment of the brain in schizophrenia is aggravated during the first 5 years of disease. PMID- 10480198 TI - Self-perceived interpersonal competence in older schizophrenia patients: the role of patient characteristics and psychosocial factors. AB - OBJECTIVE: This study compared older schizophrenia patients with normal subjects in terms of their perceived interpersonal competence. METHOD: A total of 95 middle-aged and elderly schizophrenia patients and 85 age-matched normal subjects completed the Interpersonal Competence Questionnaire. RESULTS: Patients scored significantly lower than normal subjects on initiation, provision of emotional support, and conflict management. Severity of psychiatric symptoms and other patient characteristics were examined as predictors of interpersonal competence among patients. Negative symptoms were inversely related to interpersonal competence, whereas emotional support from others and a positive appraisal coping style both yielded positive associations. CONCLUSION: These findings suggest the need for clinical interventions designed to enhance the interpersonal skills of older schizophrenia patients, particularly those with marked negative symptoms. PMID- 10480199 TI - Family interventions in schizophrenia: an analysis of non-adherence. AB - OBJECTIVE: The aim of this study was to identify baseline factors which may make it possible to predict non-adherence to prescribed treatment. METHOD: A total of 87 schizophrenic patients in a catchment area of Valencia (Spain) were randomly assigned to two family strategies. The characteristics associated with lack of adherence to the programmes were analysed in both the patients and their families. RESULTS: Older patients, those with a higher number of previous hospital admissions, those living in small households and those having relatives with little knowledge of schizophrenia at the time of the initial assessment were the main factors associated with a higher risk of dropping out of the interventions. CONCLUSIONS: The findings of this study offer some guidance on suggesting intervention strategies that might reduce drop-out rates in these types of programmes. PMID- 10480200 TI - Does the place of treatment influence the quality of life of schizophrenics? AB - OBJECTIVE: Chronically mentally ill patients in community mental health care report a better quality of life (QOL) than those in long-term hospital care, which suggests that the treatment setting per se influences their QOL. METHOD: In a region where both treatment settings are of a comparable high standard, we assessed the QOL of 96 schizophrenic patients from these two treatment settings, and the factors which most influenced their QOL. RESULTS: Community-care patients reported a better QOL than long-term hospital-care patients. However, when other factors influencing QOL were included in a regression analysis, the place of treatment was no longer significant, but rather the social support, the severity of the illness, educational level and certain illness concepts. CONCLUSION: It is probably not the place per se which influences the QOL, but apart from personal, sociodemographic and illness-related factors, the amount of social support that is provided in different settings. PMID- 10480201 TI - The association between needs and quality of life in an epidemiologically representative sample of people with psychosis. AB - The impact of meeting needs on quality of life in the severely mentally ill is investigated in this study. An epidemiologically representative sample of 133 patients meeting ICD-10 criteria for psychosis completed standardized instruments for measuring needs and quality of life. Covariance structure modelling was used to investigate the extent to which latent factors of met and unmet need were associated with latent quality of life. Patients rated about 0.7 more total (met plus unmet) needs than staff, mainly due to differences in rating unmet need. Patient ratings were more reliable than ratings by others of unmet need and quality of life. Both underlying unmet need and met need were negatively associated with underlying quality of life, but unmet need was the stronger relationship. The patient's perspective on their difficulties (especially their unmet needs) must be central to mental health care. PMID- 10480202 TI - A retrospective study of clozapine and urinary incontinence in Chinese in patients. AB - OBJECTIVE: This study investigated the incidence of clozapine-associated urinary incontinence (UI) in schizophrenic patients, the percentage of these patients with persistent urinary incontinence (PUI), and the possible factors affecting the occurrence of UI. METHOD: A total of 61 Chinese in-patients with schizophrenia (according to DSM-IV) treated with clozapine for more than 3 months were assessed retrospectively for the occurrence of UI. Patients who still had UI at the time of assessment were classified as having PUI. Patients whose UI had resolved at the time of assessment were classified as having self-limited urinary incontinence (SUI). We compared the characteristics of UI and non-UI cases and of PUI and SUI cases. RESULTS: The results showed that urinary incontinence developed at some time in 27 of 61 patients (44.3%), and that it was persistent in 15 of 61 patients (25%). There were no statistically significant differences in age, sex, clozapine dose, duration of clozapine use, duration of index admission, duration of illness, age at onset of schizophrenia, or concurrent treatment with other psychiatric medications between the UI and non-UI groups and between the PUI and SUI groups. CONCLUSION: Clozapine-associated urinary incontinence may be persistent in some patients, and it should be cautiously monitored in every patient taking clozapine. PMID- 10480203 TI - The Oslo Cognitive Training Programme. PMID- 10480204 TI - Neurofibromatosis type 1: the role of neuroradiology. AB - Neurofibromatosis type 1 is the commonest of the phakomatoses with a prevalence of 1 in 3-4000. In common with most of the other phakomatoses it is a genetic disorder with typical dermatological manifestations. Many organ systems can be affected but brain, spine, cranial nerve and peripheral nerve involvement is frequent. In this review we will describe the central nervous system manifestations of neurofibromatosis type 1 and discuss some of the controversies raised by investigating children with this disorder. PMID- 10480205 TI - Botulinum toxin A in the management of spastic gait disorders in children and young adults with cerebral palsy: a randomized, double-blind study of "high-dose" versus "low-dose" treatment. AB - The present study was performed to assess dose-response relationships of local botulinum toxin A (BtxA) treatment in children and teenagers with spastic gait due to cerebral palsy (CP) in a randomized, double-blind study employing a "high dose" (200 units Botox per leg) and a "low-dose" (100 units Botox per leg) treatment arm in 33 patients with CP. Response parameters included changes in muscle tone assessed by the Ashworth scale at knee joint, range-of-motion (ROM) measurements at knee and ankle joint, objective analysis of longitudinal gait parameters as well as subjective assessments of improvement. Patients in the "high-dose" arm received 40-80 units Botox/muscle versus 20-40 units Botox/muscle in the "low-dose" group. Patients in both treatment arms showed significant improvement of Ashworth score (p<0.001) and ROM (p<0.01), while gait analysis revealed significant increase in gait velocity (p<0.01) and stride-length (p<0.001) over baseline. Subjects in the "high-dose" group showed significantly greater improvement on objective response measurements compared to "low-dose" patients. Also, children aged 7 years or less had greater functional benefit compared to the subgroup of patients older than 7 years. Incidence and severity of side-effects were similar in both treatment groups. The present study demonstrated dose-dependent functional improvement of dynamic deformities and spastic gait pattern in children and young adults with CP treated with local injections of botulinum toxin. A dose of 200 units Botox per leg distributed to 4 or 5 muscle bellies per leg is superior compared to 100 units Botox per leg without significantly affecting the risk of side-effects. PMID- 10480206 TI - Altered cholinergic function in the basal forebrain of girls with Rett syndrome. AB - Rett syndrome (RS) is a neurodevelopmental disorder that is predominant in females and is associated with cortical atrophy, stereotyped hand movements and severe mental deficiency. Previous studies have demonstrated a significant decline in number of choline acetyltransferase (ChAT)-containing neurons throughout the forebrain of RS girls. The loss of these ChAT-positive cells may be caused by a lack of nerve growth factor (NGF). In the current study, cortical levels of NGF were normal in RS girls as compared to age-and sex-matched controls. The number of neurons within the basal forebrain that express the 75 kDa (p75) low-affinity receptor for NGF was unchanged. In contrast, the number of ChAT-positive neurons was significantly decreased. The results suggest that normal amounts of NGF are available for binding to the p75 receptor and for retrograde transport to forebrain cholinergic cells, however, these neurons do not respond by producing the ChAT protein that is necessary for the production of the neurotransmitter acetylcholine. PMID- 10480207 TI - Visual field constriction is not limited to children treated with vigabatrin. AB - Vigabatrin (GVG) is widely used in the treatment of complex partial seizures and infantile spasms. Persistent visual field constriction associated with GVG therapy in adults was reported as a rare but serious side-effect. Visual field examination in children is more difficult because of a lack of cooperation among very young or mentally handicapped patients. We performed Goldmann perimetry in 12 of 153 patients treated with GVG as mono- or as add-on therapy. The others would not cooperate, and two adolescents refused the examination. For comparison, we examined 12 age-matched patients with complex partial or generalized epilepsy who had never taken GVG. In five of 12 GVG-treated patients, and in one of the control group, we found a concentric visual field constriction. All patients were subjectively asymptomatic. The GVG-treated patients had taken the drug in combination with valproic acid (VPA) or oxcarbazepine (OCB). In four patients, GVG treatment was already stopped at the time of the ophthalmologic examination. Three patients had intracerebral lesions that could not account for the pathologic perimetric findings. The single patient from the control group with concentric visual field constriction had an absence epilepsy, treatment being performed with VPA and lamotrigine (LTG). In conclusion, GVG has a causal but not unique connection with visual field constriction in pediatric patients. PMID- 10480208 TI - Changes of neurotransmitters in the brainstem of patients with respiratory pattern disorders during childhood. AB - We examined neuropathologically and immunohistochemically the respiratory centers in the brainstem of two patients with Joubert syndrome (JS), three patients with congenital central hypoventilation syndrome (CCHS) and a patient with apneustic breathing (prolonged inspiratory pause) due to unknown etiology. Immunoreactivity (IR) of tryptophan hydroxylase (TPH) was decreased in the dorsal raphe nuclei of two patients with JS compared with age-matched controls, as well as in two patients with Dandy-Walker malformation. The two JS patients showed vermian defect and elongated cerebellar peduncles, and peculiar vascularities in the midline of the whole brainstem were also noted in one of these patients. These findings, as a whole, confirm that the midline structures of brainstem are disordered both structurally and functionally in JS, conceivably resulting in respiratory patterns and psychomotor deficits. IR of serotonin 1A receptor showed no significant changes in the medulla oblongata of these patients, however. In the parabrachial complex, IR of substance P was increased in two patients with CCHS, and one with apneustic breathing. IR of tyrosine hydroxylase was also increased in the latter. The brainstem of these patients showed reactive astrogliosis. These findings suggest preceding hypoxic episodes as well as an increased activity in the parabrachial complex which plays an important role in conducting the driving force to the medullary respiratory neurons from ascending sensory pathways. PMID- 10480209 TI - Autosomal recessive micrencephaly with simplified gyral pattern, abnormal myelination and arthrogryposis. AB - The clinical courses, neuroimaging and muscle biopsy findings of two infants born to an inbred Arab family are described. They had a syndrome of micrencephaly with simplified gyral pattern, abnormal myelin formation and arthrogryposis. Increased variation of fiber size was seen in the muscle biopsy, creatine kinase, however was normal. Large areas of muscle were replaced by adipofibrous tissue. The infants had dysmorphic features consistent with the fetal akinesia/hypokinesia sequence. The abnormalities were suggestive of microlissencephaly probably associated with a dysgenetic process in the muscles. The syndrome showed an autosomal recessive inheritance. PMID- 10480210 TI - Reduced heart rate variability in patients affected with Rett syndrome. A possible explanation for sudden death. AB - Incidence of sudden death in Rett syndrome is greater than that of the general population and cardiac electrical instability is a prime suspect cause. Our study shows that girls with Rett syndrome had significantly lower heart rate variability (marker of autonomic disarray) and longer corrected QT intervals compared with an age-matched group of healthy girls. These abnormalities increased with advancing stages of the syndrome. These findings suggest a possible role of cardiac dysfunction in the sudden death associated with Rett syndrome. PMID- 10480211 TI - Nonconvulsive status epilepticus in eyelid myoclonia with absences--evidence of provocation unrelated to photosensitivity. AB - A 10-year old girl with eyelid myoclonia with absences (EMA) in whom nonconvulsive status epilepticus developed shortly after awakening is described. A video-polygraphic recording during the status showed the characteristic eye closure provocation of eyelid myoclonia with upward deviation of the eyeballs and brief absences. Ictal EEG showed generalized polyspikes concomitant with eyelid myoclonia, while absences were accompanied by 3.5 Hz polyspike-wave complexes on EEG. This condition occurred even in total darkness as well as even after seizures precipitated by bright sunlight had been eliminated by medication. The present case suggests that the eye closure mechanism could be a more potent precipitating factor than photosensitivity in the pathophysiology of EMA. PMID- 10480212 TI - Neuroaxonal dystrophy with dystonia and pallidal involvement. AB - Infantile neuroaxonal dystrophy (INAD) is an autosomal recessive disease of infantile onset, characterised by progressive clinical course, multi-systemic involvement and widespread presence of dystrophic axons in both the central and peripheral nervous system. Clinical, neurophysiological and neuroradiological criteria of the disease are established, but the occurrence of atypical cases is known. Since the availability of molecular markers is still lacking, diagnostic evidence in vivo is provided by the presence of specific axonal lesions distally in the peripheral nerve fibres. In two children who had a protracted course of the disease with dystonic postures of the upper limbs and showed dystrophic axons following sural nerve biopsy, bilateral pallidal hypointensity was observed after T2-weighted MRI scans. These findings are consistent with iron deposition, and are usually observed in Hallervorden-Spatz syndrome (HSS), a condition which is also characterised by dystrophic axons diffusely present in the central nervous system, but without peripheral nervous system involvement. These observations raise the issue of different phenotypes of INAD, and are consistent with the existence of intermediate forms between INAD and HSS. Altered mechanisms of iron storage and transport to and from the cellular compartments may play a role in the pathogenesis of the disease. PMID- 10480213 TI - Subcortical laminar heterotopia in two sisters and their mother: MRI, clinical findings and pathogenesis. AB - MR imaging, clinical data and underlying pathogenesis of subcortical laminar heterotopia (SCLH), also known as band heterotopia, in two sisters and their mother are presented. On MR imaging a different degree of SCLH was found in all three affected family-members. The inversion recovery sequence was considered most useful in the demonstration of the heterotopic band of gray matter and the assessment of cortical thickness. The younger sister presented with epileptic seizures at the age of five months and a delayed achievement of developmental milestones. The older sister of seven years had epileptic seizures since the age of one year, and developmental delay. Their mother has only had one seizure-like episode at the age of 39. Her psychomotor development had been normal. Investigation of DNA samples of the three female family-members revealed a mutation in the X-linked doublecortin gene. Within families with band heterotopia, this gene has also been related to male family members with lissencephaly. PMID- 10480214 TI - Early onset of X-linked Emery-Dreifuss muscular dystrophy in a boy with emerin gene deletion. AB - A boy developed contractures of the Achilles tendons at 3 years and of the postcervical muscles at 7 years, although neither contractures of the elbows nor cardiac abnormality were recognized by the age of 9 years. Muscle computed tomography scanning revealed changes characteristic of muscle involvement. Emerin was not detected in the biopsied muscle, and RT-PCR and PCR-based genomic DNA analyses of the emerin gene demonstrated no amplification product in the patient. These results confirmed the diagnosis of X-linked Emery-Dreifuss muscular dystrophy (EDMD), and reinforce the necessity of molecular genetic diagnosis of the membrane protein emerin in younger patients with possible EDMD before appearance of the typical symptoms, to avoid sudden cardiac death. PMID- 10480215 TI - Landau-Kleffner syndrome and atypical benign partial epilepsy of childhood: the effectiveness of acetazolamide. PMID- 10480216 TI - A pilot study of low-dose fludarabine in membranous nephropathy refractory to therapy. AB - BACKGROUND: Lymphocytes are believed to play a role in the induction and perpetuation of membranous nephropathy. Fludarabine is a purine nucleoside analog with selective activity against both dividing and resting lymphocytes. We evaluated the tolerance, toxicity, pharmacokinetics, immunologic, and clinical effects of fludarabine in patients with membranous nephropathy in an single arm pilot study. PATIENTS AND METHODS: Eight patients with idiopathic (n = 7) or lupus (n = 1) membranous nephropathy who had failed high-dose prednisone (n = 8) and/or alkylating agents (n = 2), or cyclosporine (n = 1) were treated with 6 monthly cycles of fludarabine (cycles 1-2, 20 mg/m2/day x 2 days, cycles 3-6, 20 mg/m2/day x 3 days). Mean proteinuria was 9 g/day with a mean duration of disease of 25 months (range 12-48). Proteinuria, GFR and effective renal plasma flow were compared before and after completing the treatment. RESULTS: Seven patients completed the protocol. CD3, CD4, CD8 and B cell counts decreased by 53%, 46%, 61% and 84%, respectively, at the end of treatment and remained at lower than pretreatment levels 6 months after completing the trial. Despite lymphopenia, serum immunoglobulin levels remained unchanged. Both naive (CD45RA+) and memory CD4+ T cells (CD45RO+) were reduced (naive > memory). Proteinuria decreased by > or = 50% in 5 out of 7 patients (p = 0.11). Filtration fraction improved in all patients with decreased filtration fraction at baseline. The only side-effect observed was one episode of acute bacterial sinusitis that responded promptly to antibiotic therapy. CONCLUSION: We conclude that low-dose fludarabine treatment in patients with membranous nephropathy is well tolerated and results in significant lymphopenia involving B more than T cells. In this pilot study improvement in proteinuria and filtration rate were observed. Additional studies are required to determine the optimal dose and clinical efficacy of fludarabine. PMID- 10480217 TI - Increased serum levels of vascular endothelial growth factor in human crescentic glomerulonephritis. AB - OBJECTIVE: To investigate the serum levels of vascular endothelial growth factor (VEGF) in human crescentic glomerulonephritis (CRGN). METHODS: The serum VEGF levels in CRGN patients were compared with those in healthy subjects and in various types of glomerulonephritis. In addition, we investigated the relationship between serum VEGF levels and creatinine levels (Scr) and pathological parameters. RESULTS: The serum VEGF levels of the CRGN patients were significantly higher than in the healthy subjects and patients with MCNS, IgAN, and FGS. No correlation was found between serum VEGF levels and Scr in the RPGN patients. The serum VEGF levels in 6 CRGN patients had significantly decreased after 6 months of corticosteroid therapy. Moreover, there was a significant correlation between the serum VEGF levels and the crescent frequency or the grade of interstitial injury, and the rates of glomerular infiltration by macrophages. CONCLUSIONS: In the CRGN patients, severe glomerular and interstitial damages would result in increased serum VEGF levels and corticosteroid therapy may exert its efficacy through reduction of serum VEGF levels. PMID- 10480218 TI - Analysis of nitric oxide in the exhaled air of patients with chronic glomerulonephritis. AB - BACKGROUND: Nitric oxide (NO) plays an important role in renal hemodynamics and function. Although production of NO in the glomeruli has been found to be increased in animal models of glomerulonephritis, it remains unclear whether its endogenous production is enhanced in patients with chronic glomerulonephritis (CGN). SUBJECTS AND METHODS: We measured NO output in exhaled air as an indicator of its local production in the lungs and plasma and urinary nitrite plus nitrate (NO2-/NO3-) levels as indicators of its production in the whole body in 21 patients with CGN in 31 healthy controls. RESULTS: The patients exhaled higher concentrations of NO (29.5 +/- 1.4 vs. 18.7 +/- 1.0 parts per billion (ppb), mean +/- SEM, p < 0.0001) and exhaled NO output was also higher than in controls (166.6 +/- 6.8 vs. 95.5 +/- 5.6 nl/min/m2, p < 0.0001). Plasma NO2-/NO3- concentrations were also significantly greater in the patients than in the controls (81.6 +/- 7.2 vs. 41.1 +/- 4.3 micromol/l, p < 0.001). In patients with CGN, exhaled NO output correlated negatively with creatinine clearance (r = 0.62, p < 0.05). Oral administration of prednisolone (60 mg/day) for two weeks did not significantly affect the exhaled NO output in the patients (160 +/- 7 vs. 200 +/- 30 nl/min/m2, p = NS) despite a decrease in urinary protein excretion (12.0 +/- 2.9 vs. 1.4 +/- 0.6 g/day, p < 0.01). CONCLUSION: These findings suggested that endogenous NO production is increased in patients with CGN. Increased endogenous NO production may play some pathophysiological role in these patients. PMID- 10480220 TI - The changing pattern of glomerulonephritis in Singapore over the past two decades. AB - This study reviews the pattern of glomerulonephritis (GN) in Singapore over the past 2 decades. In the earlier decade the pattern was typical of most Asian countries with mesangial proliferative GN (Mes GN) (56%) as the most common form of primary GN including the nephrotic syndrome (40%). In the 2nd decade the pattern undergoes a change. Though Mes GN is the commonest primary GN (42%), the commonest form of nephrotic syndrome is now minimal change disease (30%) with Mes GN decreasing to 25% among all primary nephrotic syndromes. Both minimal change and focal global sclerosis account for 50% of steroid/cyclophosphamide responsive GN today. Membranous GN though still uncommon, has increased from 3% (1st decade) to 6% (2nd decade) (p < 0.01). IgA nephritis is still the commonest primary GN occurring in Singapore (42% of all primary GN in the 1st decade and 45% in the 2nd decade). The present pattern of GN in Singapore, though, still predominantly Asian with the preponderance of mesangial proliferative GN with a relatively low incidence of membranous GN contrasts with the pattern in the West where membranous GN is the commonest form of primary GN. Even the incidence of FSGS has not increased as in the West where there is a rising incidence. The underlying basis for most GN in Singapore as in other Asian countries and elsewhere is antigen-driven: infective antigen as well as food or other allergens. PMID- 10480219 TI - Concealed hypothyroidism in patients with myeloperoxidase antineutrophili cytoplasmic autoantibodies- (MPO-ANCA) positive renal disease. AB - AIM: To elucidate the relationship between thyroid disorders and myeloperoxidase (MPO) antineutrophili cytoplasmic autoantibodies-(ANCA) positive renal disease, we investigated the prevalence of thyroid diseases in patients with them. SUBJECTS AND METHODS: The subjects consisted of 10 patients (6 males and 4 females) diagnosed as having MPO-ANCA-positive renal disease in our hospital. To evaluate thyroid function, serum thyroid-stimulating hormone (TSH) and free thyroxine (FT4) levels were measured by a immunoradiometric assay (IRMA) and a radioimmunoassay (RIA), respectively. Thyroid autoantibodies against microsomal antigen and thyroglobulin were measured by means of particle agglutination. In addition, antibodies against thyroid peroxidase (TPO-Ab) and purified thyroglobulin (Tg-Ab) were also determined by RIA. To assess the disease activity, we employed Birmingham Vasculitis Activity Score (BVAS). RESULTS: The results were that 4 out of 10 patients (40%) were accompanied by hypothyroidism, including 2 patients with subclinical hypothyroidism and I during thyroid hormone replacement therapy. The BVAS of the hypothyroidism group (group A) was significantly lower (p < 0.05) than that of non-hypothyroidism group (group B), 18.5 +/- 9.0 (mean +/- SD) versus 24.2 +/- 16.6. MPO-ANCA titers were not significant between the two groups, 376.8 +/- 316.8 EU versus 311.5 +/- 203.6 EU. CONCLUSION: Thyroid disorder, especially hypothyroidism would not be a rare complication in patients with MPO-ANCA-positive renal disease. We should concentrate more on thyroid lesion in them. PMID- 10480221 TI - Growth hormone, insulin-like growth factors and their binding proteins in adult hemodialysis patients treated with recombinant human growth hormone. AB - BACKGROUND: Growth deficiency and malnutrition in uremic children are often caused by malfunction of the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis and can be corrected by treatment with GH. The purpose of this study was to evaluate the levels of GH, IGF-I and II and their binding proteins compared to changes in body composition in adult, enfeebled, uremic patients in chronic hemodialysis (HD), treated for 6 months with recombinant human growth hormone (rhGH). METHODS: 31 patients were included in a controlled, randomized, double-blinded study using either 4 IU/m2/day of rhGH or placebo injected subcutaneously every evening for 6 months. RESULTS: Fasting levels of GH were normal at start and increased significantly from 2.2 to 13.5 microg/l (p = 0.01) within the first 4 months of rhGH treatment. Before treatment IGF-I was at the upper limit of normal range (130 to 220 microg/l) in both groups, and it increased significantly from 213 to 348 microg/l (p = 0.01) during rhGH treatment. IGF-II was above the normal range in both groups, and remained unchanged throughout. IGFBP-1 decreased in the rhGH-treated group from 53.1 to 24.7 microg/l (p = 0.004), while IGFBP-3 increased from 5620 to 7100 microg/l (p = 0.004). The molar ratio of IGF-I/IGFBP-3 increased significantly from 14 to 25% (p = 0.01), while the ratio decreased in the placebo group (p = 0.01). During the treatment with rhGH the patients increased their lean body mass (= muscle mass) by a median of 3.18 kg (range 0.82 to 5.12 kg) (p = 0.0001) while their fat mass decreased by a median of 3.33 kg (range 0.18 to 5.82 kg) (p = 0.004). Total body mass (= weight) remained stable. No significant changes were observed in the placebo group. CONCLUSION: The baseline GH and IGF-I concentrations were normal in malnourished HD patients. When treated with rhGH in a dosage as used in growth retarded uremic children, IGF-I increased to the levels seen in acromegalic persons. IGF-I increased more than IGFBP-3 whereby its biological activity obviously improved. This was reflected in an increased muscle mass and a decreased fat mass. The rhGH treatment was well tolerated. PMID- 10480222 TI - The pattern of intracellular free amino acids in granulocytes from hemodialysis patients change to an oral protein supplement (granulocyte amino acids after protein in HD patients). AB - MATERIALS AND METHODS: The concentrations of free intracellular amino acids in granulocytes and plasma amino acids, normalized protein nitrogen appearance rate, serum insulin-like growth factors, plasma proteins, anthropometric and bioimpedance measurements were determined before and after an oral protein supplement in 19 stable patients on maintenance hemodialysis in a randomized, double-blind placebo-controlled study with crossover after 3 months. The hemodialysis patients were well-nourished with an ideal body weight of 91% after both protein supplementation and after placebo. RESULTS: After protein supplementation (7.8 g/d) the intracellular concentration of valine, isoleucine, threonine and tyrosine and the valine/glycine and tyrosine/phenylalanine ratios in the cells were significantly increased (p < 0.05). In contrast, the concentrations of plasma amino acids, serum insulin-like growth factors, and plasma proteins and body weight and anthropometric and bioimpedance measurements were unchanged. Dialysis efficiency was unchanged throughout the study. CONCLUSIONS: The present study supports the conclusion that protein supplementation to well-nourished hemodialysis patients does not improve the nutritional status measured by plasma proteins, body weight, anthropometric and bioimpedance measurements. The increase in intracellular amino acid concentrations indicates better cellular nutrition and metabolic control. PMID- 10480223 TI - Increased plasma levels of mature form of adrenomedullin in patients with chronic renal failure. AB - BACKGROUND: Adrenomedullin (AM), a novel vasodilator peptide, is produced by C terminal amidation reaction of AM-glycine. AM-glycine, an intermediate form of AM (iAM), is processed from pro AM. AM circulating in the human blood stream was found to consist of an amidated mature form (mAM) and iAM. Biological activity is exerted only by mAM. METHODS: To investigate the pathophysiological role of mAM in renal disease, we measured plasma concentrations of mAM as well as total AM (tAM), representing both mAM and iAM, in patients with various renal diseases. In addition, plasma ANP level was measured in all patients. RESULTS: The concentrations of plasma mAM in renal failure with dialysis (2.1 +/- 0.2 fmol/ml, mean +/- SEM) and without dialysis (1.2 +/- 0.2) were significantly (p < 0.05) higher than those in control group (0.5 +/- 0.1). However, the plasma ANP level was increased only in renal failure patients with dialysis. Plasma mAM levels were significantly correlated positively with serum creatinine levels and negatively with hematocrit. No significant difference was noted in the ratio of mAM/tAM between renal failure patients and healthy subjects. CONCLUSION: These results suggest that plasma mAM is increased in renal failure in relation to deterioration of renal function, while the amidation process of AM seems to be unaffected in patients with renal failure. PMID- 10480225 TI - Network modeling of the convective flow and diffusion of molecules adsorbing in monoliths and in porous particles packed in a chromatographic column. AB - A cubic lattice network of interconnected pores was constructed to represent the porous structure existing in a monolith (continuous bed) or in a column packed with porous chromatographic particles. Expressions were also constructed and utilized to simulate, through the use of the pore network model, the intraparticle interstitial velocity and pore diffusivity of adsorbate molecules in porous chromatographic particles or in monoliths under retained and unretained conditions. The combined effects of steric hindrance at the entrance to the pores and frictional resistance within the pores, as well as the effects of pore size, pore connectivity, nT, of the porous network, molecular size of adsorbate and ligand (active site), and the fractional saturation of adsorption sites (ligands), have been considered. The results for the adsorption systems studied in this work, indicate that the obstruction effects on the intraparticle interstitial velocity, due to (a) the thickness of the immobilized layer of active sites and (b) the thickness of the adsorbed layer, are small and appear to be insignificant when they are compared with the very significant effect that the value of the pore connectivity, nT, has on the magnitude of the intraparticle interstitial velocity. The effective pore diffusion coefficient of the adsorbate molecules was found to decline with increasing molecular size of ligand, with increasing fractional saturation of the active sites or with diminishing pore size, and with decreasing pore connectivity, nT. The results also show that the magnitude of the interstitial fluid velocity is many times larger than the diffusion velocity of the adsorbate molecules within the porous adsorbent particles. Furthermore, the results clearly show that the intraparticle interstitial velocity and the pore diffusivity of the adsorbate increase significantly as the value of the pore connectivity, nT, of the porous medium increases. The results of this work indicate that the pore network model and the expressions presented in this work, could allow one, for a given porous adsorbent, adsorbate, ligand (active site), and interstitial column fluid velocity, to determine in an a priori manner the values of the intraparticle interstitial velocity and pore diffusivity within the monolith or within the porous adsorbent particles as the fractional saturation of the active sites changes. The values of these transport parameters could then be employed in the macroscopic models that could predict the dynamic behavior, scale-up, and design of chromatographic systems. The theoretical results could also have important implications in the selection of a ligand as well as in the selection and construction of an affinity porous matrix. PMID- 10480224 TI - Mechanical complications in chronic peritoneal dialysis. AB - BACKGROUND: A significant number of complications and catheter failures in CAPD are due to mechanical problems. METHODS: We reviewed 105 consecutive peritoneal dialysis catheters, generally implanted by blind insertion technique, in 80 patients over an 11-year period. 137 mechanical problems were collected and classified according to their etiologies into the following: those related to the insertion procedure (n = 21; 15.3%), those related to the presence of dialysate in peritoneum (n = 33; 24%), catheter-related malfunction (n = 28; 20.4%), secondaries to abdominal events (n = 3; 0.2%), catheter accidents (n = 37; 26.2%) and others (n = 15; 11%). RESULTS: Removal of the catheter was performed in 86 of 105 catheters: mechanical complications accounted for 19 of them (21.8%) and 9 patients were definitively transferred to hemodialysis due to this problem. The most frequent complications were catheter accidents, although only a few of them required catheter removal. CONCLUSION: The majority of the catheter removals were due to catheter displacement (31.5%), this problem being more frequent in straight than in Swan Neck catheters. PMID- 10480226 TI - Retention behavior of very large biomolecules in ion-exchange chromatography. AB - Hepatitis B virus surface antigen (HBsAg) particles were efficiently adsorbed (retained) on a Sulfate-cellulose (S-C) bead column, and then desorbed with sodium chloride solutions (0.5-3.0 M). The HBsAg particles were not efficiently retained onto either sulfopropyl-agarose (SP-A) or quaternary amine-agarose (Q-A) at pH 4.5, 6 and 8. The size-exclusion curve showed that proteins of molecular mass higher than ca. 20,000 cannot penetrate into the pores of S-C beads. The dynamic binding capacity (DBC) values of lysozyme (ca. 7 mg/ml-gel) and of gamma globulin (ca. 3 mg/ml gel) for S-C did not depend on the flow velocity while the DBC of gamma-globulin for SP-A decreased sharply with an increase in flow velocity. These results indicated that very large molecules are adsorbed only onto the surface of S-C, which resulted in fast adsorption-desorption rates although the equilibrium adsorption capacity is lower than conventional porous gel beads. Because of the rapid adsorption rate, the DBC values of gamma-globulin for S-C at high flow-rate regions are similar to those for SP-A. Bovine serum albumin was not adsorbed onto S-C. As this can not be explained by a simple electrostatic interaction mechanism, molecular recognition of S-C might be different from the agarose-based ion-exchange beads. PMID- 10480227 TI - Ion-exchange chromatography of proteins near the isoelectric points. AB - The retention and the resolution of beta-lactoglobulin A and B (LgA, LgB) were investigated with various ion-exchange chromatography media. The number of sites involved in the retention (adsorption) decreased as the mobile phase pH approached the isoelectric points pI (=5.1-5.2). However, even at pH 5.2 both LgA and LgB were retained on anion- and cation-exchange chromatography columns. The separation (resolution) of LgA and LgB became better when the pH approached the pI in anion-exchange chromatography columns where the number of adsorption site values are small (ca. 2-3). The two proteins were not separated on cation exchange chromatography columns. Factors affecting the resolution and the retention near the pI were discussed. PMID- 10480228 TI - Characterization of unstable ion-exchange chromatographic separation of proteins. AB - Very fine separation of proteins by stepwise elution ion-exchange chromatography is very often a unstable process. To characterize the unstability of such processes the elution volume variations were examined by the model equation which contained the ion-exchange capacity and the number of adsorption sites. The data needed for the model calculation were obtained from gradient elution experiments. As a model separation system stepwise elution of a model protein (beta lactoglobulin) near the isoelectric point on a weak cation-exchange chromatography column was chosen. The elution volume varied significantly with a small change in the ion-exchange capacity. It was found that the ionic strength of the elution buffer must be adjusted in order to compensate a change in the elution volume due to the ion-exchange capacity variations. The ionic strength and the pH of the elution buffer were also found to be important variables affecting the elution volume. In this model separation system, it was indicated that the pH should be within +/-0.1 unit and the ionic strength within +/-0.002 mol/l in order to meet the criteria (+/-5% elution volume variation). It is recommended that gradient elution data be obtained for predicting elution volume variations in stepwise elution. By using the gradient elution data the process diagnosis can be performed, and the important information on the process stability can be obtained. PMID- 10480229 TI - Comparison between the isocratic and gradient retention behaviour of polypeptides in reversed-phase liquid chromatographic environments. AB - The isocratic and gradient elution behaviour of beta-endorphin and glucagon, two polypeptides known to exist in amphipathic alpha-helical conformations in lipophilic environments, have been examined under reversed-phase high-performance liquid chromatographic (RP-HPLC) conditions with low pH, aquo-acetonitrile mobile phases. The effects of changes in the volume fraction, psi, of the organic solvent modifier and temperature, T, on the magnitudes of the S and log k(o) values of these two polypeptides, obtained from the plots of logarithmic capacity factor (log k') vs. psi using isocratic elution conditions have been determined. These data have then been compared to the corresponding S and log k(o) values, obtained from the plots of logarithmic median capacity factor (log k) versus the median volume fraction of the organic solvent modifier (psi) derived from the linear gradient elution data, using the same n-butyl silica sorbent and related aquo-acetonitrile mobile phase conditions. As apparent from these studies, substantial differences occur in the temperature-dependent trends and magnitudes of the corresponding S and S values, or the log k(o) and log k(o) values, when these parameters are derived from experimental data acquired by these two different elution methods. Moreover, when gradient elution data for beta endorphin and glucagon are utilised, the extrapolated values of the intercept and slope of the plots of log k vs. 1/T (corresponding to an apparent change in the median enthalpy of association, deltaH(o)assoc, or an apparent change in the median entropy of association, deltaS(o)assoc) substantially deviated from the values obtained for the thermodynamic parameters, deltaH(o)assoc and deltaS(o)assoc, derived from the log k' vs. 1/T plots using the corresponding isocratic data. These findings thus have important implications for biophysical and thermodynamic investigations when gradient elution data are employed to assess the molecular basis of the interaction of polypeptides with non-polar ligates. PMID- 10480230 TI - Use of surface plasmon resonance for studies of protein-protein and protein phospholipid membrane interactions. Application to the binding of factor VIII to von Willebrand factor and to phosphatidylserine-containing membranes. AB - The surface plasmon resonance phenomenon is used for real time measurements of protein-protein and protein-membrane interactions. In the present study two surface plasmon resonance-based binding assays permitting study of the interaction of coagulation factor VIII (fVIII) with von Willebrand factor (vWf) and phospholipid have been developed. These interactions of fVIII are required for maintenance of fVIII concentration in circulation and for the assembly of the functional factor Xase complex, respectively. With these binding assays, the role of the light chain (LCh) in fVIII binding to vWf and to immobilized phospholipid monolayers and intact vesicles containing 25% phosphatidylserine (PS) and 4% PS was examined. The finding that Kd of LCh binding to vWf (3.8 nM) is 9.5 times higher than that of fVIII (0.4 nM), indicates that the heavy chain (HCh) is required for the maximal affinity of fVIII for vWf. In contrast, affinities of LCh for 25/75 PS/phosphatidylcholine (PC) monolayers and 4/76/20 PSPC phosphatidylethanolamine (PE) vesicles are similar to that of fVIII, indicating that LCh is solely responsible for these interactions. It was also examined how removal of the acidic region affects the binding affinity of the remaining part of LCh for vWf and phospholipid. It was demonstrated that the loss of the LCh acidic region upon thrombin cleavage leads to an 11 and 160-fold increase in the dissociation rate constant (k(off) value) and a 165 and 1500-fold increase in the Kd value of the binding of fVIII fragment A3-C1-C2 to vWf compared to that of LCh and fVIII, respectively. In contrast, the binding affinity of A3-C1-C2 for PS containing membranes was 8-11-fold higher than that of LCh. Possible conformational change(s) in C2 domain upon removal of the acidic region were studied using anti-fVIII monoclonal antibody NMC-VIII/5 with an epitope within the C2 domain of LCh as a probe. The determined lower binding affinity of A3-C1 C2 for NMC-VIII/5 immobilized to a sensor chip than that of LCh, indicates that these conformational changes do occur. PMID- 10480231 TI - Large-scale membrane adsorbers. AB - A new large-scale membrane adsorber system for rapid separation of biomolecules based on a hollow cylindrical module geometry in unitized construction and combined parallel and serial connection of modules is presented. The effectiveness of the concept is demonstrated by breakthrough curves on a 21-m2 pilot plant with hemoglobin as a model substance. Scale-up to the order of 100 m2 and above is easily performed by combining modules in parallel for increasing capacity, and in series for optimizing performance. An estimation of productivity based on these data is presented. Preliminary results of separations by linear salt gradient elution are also given. PMID- 10480232 TI - Ion-exchange chromatography separation of the detergent and the solvent from immunoglobulins after solvent-detergent treatment. AB - For inactivation of lipid-enveloped viruses during the immunoglobulin production, the solvent-detergent (S/D) method was applied. Tri-n-butyl phosphate (solvent) and Triton X-100 (detergent) were removed from S/D treated immunoglobulins by ion exchange chromatography on Q-Sepharose Fast Flow (FF). During the chromatographic procedure immunoglobulins remained bound on a Q-Sepharose FF, whereas solvent and detergent were eluted by washing with starting buffer. Elution of immunoglobulins was achieved by increasing the ionic strength of the starting buffer. The final immunoglobulin preparation contained less than 10 microg/ml of Triton X-100 and less than 2 microg/ml tri-n-butyl phosphate. It was confirmed that the S/D procedure did not cause a significant change in polymers and specific antibodies content. Immunoglobulin classes were also not affected by the same procedure. PMID- 10480233 TI - Removal of detergent and solvent from solvent-detergent-treated immunoglobulins. AB - The solvent-detergent (S/D) method was applied for inactivation of lipid enveloped viruses during the production of immunoglobulins. Amberlite XAD-7 resin was used for removal of solvent (tri-n-butyl phosphate, TnBP) and detergent (Triton X-100) after the performed S/D inactivation procedure. The S/D reagents from the immunoglobulin preparation were adsorbed on Amberlite XAD-7, while immunoglobulins passed through the column and retained their biological activity. Using the method developed here, the final immunoglobulin preparation contains less than 1 ppm of Triton X-100 and less than 2 ppm TnBP. PMID- 10480234 TI - Biomembrane-affinity centrifugal analyses of solute interactions with membrane proteins. AB - We have developed a rapid centrifugal method for analyzing solute interactions with membrane proteins in cytoskeleton-depleted membrane vesicles or proteoliposomes sterically immobilized in Superdex 200 gel beads. The size and density of the gel beads allow fast sedimentation in a bench-top centrifuge. Biospecific interactions of cytochalasin B and D-glucose with the human red cell glucose transporter, Glut1, were analyzed. The binding constants and the molar ratio of inhibitor sites per protein monomer agreed well with recent results obtained by frontal affinity chromatography. PMID- 10480235 TI - Sandwich-type immunoassay with an affinity column coupled to anti-peptide antibodies. AB - A new sandwich-type immunoassay method utilizing an anti-peptide antibody column and a fluorescence-labeled antibody (FLA) was proposed and applied to the measurement of the concentrations of insulin and insulin B-chain. In this method an immunoadsorbent coupled with an anti-peptide antibody was packed in a small column. A target protein was adsorbed on the anti-peptide antibody in the column, coupled with the FLA as a secondary antibody in sandwich manner and then eluted by an eluent containing an antigen peptide to the anti-peptide antibody. The concentration of the target protein was determined form the fluorescent intensity of the eluted FLA. It was possible to determine the concentrations within 30 min, and the higher sensitivity was obtained in comparison with the direct method using an immunoaffinity column. PMID- 10480236 TI - New ligand, N-(2-pyridylmethyl)aminoacetate, for use in the immobilised metal ion affinity chromatographic separation of proteins. AB - A new chelating compound has been developed for use in the immobilised metal ion affinity chromatographic separation of proteins. The tridentate ligand, sodium N (2-pyridylmethyl)aminoacetate (carbpyr), 1, was prepared via a one-step synthesis from 2-picolylamine, 3 and then immobilised onto Sepharose CL-4B through the epoxide coupling procedure. The binding behaviour of the resulting IMAC sorbent, following chelation with Cu2+ ions to a density of 152 micromol Cu2+ ions/g gel was characterised by frontal analysis experiments using horse heart myoglobin (HMYO) at pH 7.0 and pH 9.0. From the derived isotherms, the adsorption capacity, q(m), for the binding of HMYO to immobilised Cu2+-N-(2-pyridylmethyl)aminoacetate (im-Cu2+-carbpyr)-Sepharose CL-4B at these pH values was found to be 1.92 and 1.91 micromol/g sorbent, respectively, whilst the dissociation constants K(D) were 0.0092 x 10(-6) M and 0.0062 x 10(-6) M at pH 7.0 and pH 9.0, respectively, indicating that the HMYO-im-Cu2+-N-(2-pyridylmethyl)aminoacetate complex was more stable under alkaline conditions, although the binding capacity in terms of micromol protein/g gel remained essentially unchanged. The selectivity features of the im-Cu2+-carbpyr-Sepharose CL-4B sorbent were further characterised in terms of the binding properties with several human serum proteins at pH 5.0, pH 7.0 and pH 9.0. PMID- 10480237 TI - Histidines in affinity tags and surface clusters for immobilized metal-ion affinity chromatography of trimeric tumor necrosis factor alpha. AB - In order to achieve efficient IMAC (immobilized metal-ion affinity chromatography) purification of tumor necrosis factor alpha (TNF-alpha) and its analogs by a common chromatographic procedure, we tested four histidine-rich affinity tags attached to the N-termini of the trimeric TNF-alpha molecule. Using low cultivation temperature and appropriate protease deficient E. coli strains, it was possible to obtain intact, full-length proteins with NHis2Xa and HisArg tags, which could be purified to over 95% purity in a single step. However, in comparison to model proteins bearing a surface histidine cluster, accumulation of the histidine-tagged proteins in E. coli was significantly reduced, even in protease deficient strains. In addition, the histidine tagged TNF-alpha proteins never displayed good chromatographic behavior, which was otherwise easily achieved with model proteins. Although the most popular hexa-histidine tag is generally recognized as very convenient for single step isolation of monomeric proteins, our results with trimeric TNF-alpha indicate that oligomeric proteins may require further optimization of the tag, with respect to its length, composition, and location. Histidines, relatively rigidly inserted in the structure, as in our model proteins, display superior chromatographic characteristics vis a vis flexible tags with the same total number of histidines. PMID- 10480238 TI - Quantitative fast fractionation of a pool of polyclonal antibodies by immunoaffinity membrane chromatography. AB - A new affinity method for the direct quantitative analysis of monospecific anti peptide immunoglobulins (antibodies) and, simultaneously, their semi-preparative isolation from blood serum of the immunized animals has been developed. Immunoaffinity discs based on macroporous poly(glycidyl methacrylate-co-ethylene dimethacrylate) were used as the supporting stationary phase. The specifically prepared synthetic peptides with biological activity imitating that of the immunoglobulin binding sites of various proteins were used as the selective ligands instead of native proteins. These ligands were immobilized by a single step reaction that involves epoxy groups located on the pore surface of the porous polymer disc with amine groups of the peptide molecules. A spacer between biospecific ligands and the linking site was not required to achieve good separation. These novel immunosorbents characterized by large binding capacity are well suited for high throughput screening. Dissociation constants of the peptide-antibody complexes calculated from the experimental adsorption isotherms confirm the excellent selectivity of the proposed separation method. The discs were used in a single step enrichment of antibodies both from precipitated blood fraction and crude blood serum of immunized animals. The quantitative data of the immunoaffinity disc chromatography were compared to those obtained by an enzyme linked immunosorbent assay. Gel electrophoresis was also used to demonstrate the high degree of purity of the final product. In contrast to typical techniques that involve proteins, this immunoaffinity approach allows for the first time direct determination of concentration of specific antibodies using the immunosorbent prepared from the short peptide molecules immobilized on the internal surface of reactive porous polymer discs. PMID- 10480239 TI - Oriented immobilization of chymotrypsin by use of suitable antibodies coupled to a nonporous solid support. AB - In order to eliminate the kinetic limitation of chymotryptic hydrolysis of proteins due to diffusion, nonporous hydroxyalkyl methacrylate solid support was developed and used for oriented immobilization of chymotrypsin by means of suitable polyclonal antibodies. Nonporous microspheres were prepared by dispersion copolymerization of 2-hydroxyethyl methacrylate and ethylene dimethacrylate in an alcohol-toluene mixture stabilized with cellulose acetate butyrate. The resulting particles were 1.2 microm in diameter and possessed narrow size distribution. After modification with adipic acid dihydrazide they contained 2 micromol of reactive groups available for coupling of anti chymotrypsin antibodies. Prepared immunosorbent adsorbed 166.7 microg of chymotrypsin per 1 g of dry carrier. Immobilized chymotrypsin retained practically 100% of its native proteolytic activity. Kinetic parameters of catalysis by chymotrypsin immobilized via this way were improved due to the good steric accessibility of the enzyme active site for high-molecular-mass substrates, when digestion of proteins in batch experiments was used. PMID- 10480240 TI - Affinity purification of Schistosoma japonicum glutathione-S-transferase and its site-directed mutants with glutathione affinity chromatography and immobilized metal affinity chromatography. AB - A C-terminally polyhistidine-tagged protein of Schistosoma japonicum glutathione S-transferase, named as SjGST/His, and its Cys85-->Ser, Cys138-->Ser, and Cys178- >Ser site-directed mutants were prepared and highly expressed in Escherichia coli. Both immobilized metal affinity chromatography (IMAC) and glutathione (GSH) affinity chromatography were used to purify these four enzymes. All of them were purified with equal efficiency by Ni2+-chelated nitrilotriacetic acid agarose gel, but not by GSH Sepharose 4B gel. The protein amounts of wild-type and Cys85- >Ser enzymes purified by the latter gel were three to seven-fold greater than those of the other two enzymes purified by the same gel, while their specific activities were two-fold lower, presumably because of the occurrence of noncovalent aggregation. Both purification methods yielded highly pure enzymes, while there were minor amounts of inter- and intra-disulfide forms in the IMAC purified enzymes except for the Cys85-->Ser mutant. Addition of dithiothreitol to GSH-affinity purified enzymes shifted all of their mass spectra of matrix assisted laser desorption/ionization-time of flight mass spectrometry toward low molecular-mass regions, while addition of GSH to IMAC purified enzymes shifted the spectra toward high molecular-mass regions. The shift values of wild-type enzyme were larger than those of the three mutants, indicating that the Cys85, Cys138, and Cys178 residues were S-thiolated by GSH during the GSH-affinity purification. This result was confirmed by isoelectric focusing. These findings suggest that IMAC is more efficient than the conventional GSH-affinity system for the purification of SjGST/His enzyme, especially for its mutants and fusion proteins. PMID- 10480241 TI - Affinity chromatography of human estrogen receptor-alpha expressed in Saccharomyces cerevisiae. Combination of heparin- and 17beta-estradiol-affinity chromatography. AB - Estrogen receptor-alpha is a member of the nuclear hormone receptor superfamily and is considered as a very important regulatory protein. Human estrogen receptor alpha has been cloned into Saccharomyces cerevisiae as a fusion to ubiquitin and expression is controlled by a metallothionin promotor. Pilot scale quantities of receptor have been produced by a yeast strain transformed with expression plasmid YEpE13 [Graumann et al., J. Steroid Biochem. Mol. Biol. 57 (1996) 293] in a 14 l stirred tank reactor. The yeast extract contained 2-4 pmol of receptor protein per mg total protein. A purification scheme has been developed using heparin affinity chromatography combined with affinity chromatography with immobilized 17beta-estradiol 17-hemisuccinate. Heparin-affinity chromatography was very efficient to remove host cell protein. Accompanying proteins that stabilize unoccupied receptor have not been dissociated during elution. The receptor could be purified 5-10-fold in ligand-free state. In contrast to previous reports, we did not find a difference of the binding affinity of liganded and unliganded receptor for heparin immobilized onto Sepharose. The unoccupied receptor could be further purified 100-fold with ligand-affinity chromatography using 17beta estradiol 17-hemisuccinate-bovine serum albumin-Sepharose. The receptor could be kept in its native state, although saturated with 17beta-estradiol. The purification sequence allows an efficient production of receptor. Further improvement of productivity can be only accomplished by increasing the expression level. PMID- 10480242 TI - Use of high-resolution techniques for the characterization of clotting factor VIII. AB - Dealing with the structural characterization of clotting factor VIII (FVIII) requires the application of several high-resolution analytical techniques. Besides the analytical point of view, a detailed knowledge of FVIII structure, production and therapeutic application is necessary. This review gives an overview of most of the currently applied analytical methods and how they deal with the complex analytical problem, investigating FVIII in a sample matrix containing large amounts of accompanying plasma proteins. PMID- 10480243 TI - Solid-phase synthesis, isolation and analysis of a mouse protein, the macrophage migration inhibitory factor. AB - Murine macrophage migration inhibitory factor (mMIF) is an Mr 12,500 protein composed of all natural amino acid residues. Using the fluorenylmethoxycarbonyl chemistry for solid-phase peptide synthesis (SPPS) under special conditions, a stepwise approach was very successful leading to a crude product in unexpected high purity. After RP-HPLC isolation, a little mass difference of -12 u was determined for the received protein by matrix-assisted laser desorption ionization MS and ion spray MS and the failure sequence [Ser15]-mMIF identified by Edman degradation. The total solid-phase synthesis was repeated in the stepwise manner under the same conditions leading to the expected mMIF protein in high purity, which was confirmed by different analytical methods. Our results shows the reproducibility of our SPPS approaches to proteins and point out the importance of high-resolution mass spectrometry for a rapid and accurate analysis of such biopolymers. PMID- 10480244 TI - Strategy for identifying circulating fragments of insulin-like growth factor binding proteins in a hemofiltrate peptide bank. AB - A differentiated strategy was established to isolate circulating forms of the six human insulin-like growth factor binding proteins (IGFBPs). As starting material we used our peptide bank, a comprehensive blood plasma peptidoma generated from human blood filtrate. The peptides were initially identified in the fractions of the hemofiltrate peptide bank by their immunoreactivity, their capacity to bind the insulin-like growth factors (IGFs), and their molecular masses determined by polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS). Fractions revealing both immunoreactivity and IGF-binding capacity were analyzed by direct sequencing of immunoreactive bands from a Coomassie-stained gel. Further purification of the IGFBP peptides was performed by consecutive chromatographic steps guided by sensitive MALDI-MS. Using this strategy, different fragments of IGFBP-3, -4, and 5 were identified and a fragment of IGFBP-4 was purified to homogeneity. PMID- 10480245 TI - Purification of two low-molecular-mass serine proteinase inhibitors from chicken liver. AB - Two serine proteinase inhibitors, designated clTI-1 and clTI-2 were purified from livers of chickens to apparent homogeneity by a combination of ethanol-acetone fractionation, gel filtration and ion-exchange chromatography on CM-cellulose and Mono S columns. The inhibitor clTI-1 is a single polypeptide chain, low-molecular mass protein (Mr about 6200), very stable to heat and ethanol. It inhibits chicken, porcine and bovine trypsins as well as human plasmin. The second protein, clTI-2 of Mr 17,000 was shown to be a very effective inhibitor of both trypsins and human cathepsin G. Since both inhibitors are sensitive to arginine modification with phenylglyoxal it is assumed that this amino acid residue is present at the P1 position of the reactive site peptide bond. The N-terminal amino acid sequence of 28 residues of clTI-2 (SVDVSKYPSTVSKDGRTLVACPRILSPV) revealed a high homology of this protein to the third domain of the chicken ovoinhibitor, whereas, the clTI-1 (APPAAEKYYSLPPGAPRYYSPVV) has some sequence identity to a fragment of the human inter-alpha-trypsin inhibitor. PMID- 10480246 TI - Purification and partial characterization of the pancreatic proteolytic enzymes trypsin, chymotrypsin, and elastase from the chicken. AB - The purpose of this work was to isolate, purify and partially sequence trypsin, chymotrypsin and elastase from the chicken pancreas. The extraction of the pancreatic zymogens with 0.5 M CaCl2 at pH 7.5 for 9 h appeared to be most effective in obtaining maximum recovery of the three enzymes. The sequential Cucurbita maxima trypsin inhibitor I/bovine pancreas trypsin inhibitor/soybean trypsin inhibitor affinity chromatography gave the best result for the isolation of trypsin, chymotrypsin and elastase, respectively, from the same extract. For each proteinase, multiple form of enzymatic activity could be observed after gel electrophoresis and each form was further purified on an ion-exchange column. The N-terminal amino acid sequence of trypsin and chymotrypsin showed homologies with the bovine enzymes whereas elastase showed homologies with the porcine enzyme. The molecular mass of trypsin, chymotrypsin and elastase were estimated to be 23,500, 25,700 and 25,000, respectively, which are values close to those in mammalian species. Although some kinetic constants (Km and k(cat)/Km) appeared different from those observed in other species, the pH dependent enzymatic activities were similar to those reported in other animal species. PMID- 10480247 TI - High concentrations of sodium chloride facilitate the use of immobilized chymotrypsin for separating virgin forms of specific trypsin inhibitors. AB - It has been shown that specific trypsin inhibitors exhibit also antichymotrypsin activity in the presence of high NaCl concentrations. Taking advantage of this phenomenon a simple procedure of separation of the virgin forms of trypsin inhibitors from squash seeds and porcine pancreas (Kazal) was elaborated. In a typical experiment the inhibitor sample was loaded onto immobilized chymotrypsin equilibrated with 5 M NaCl at pH 8. After washing out unadsorbed material the virgin forms of inhibitors could be eluted either with water, buffer pH 8.0 or 0.02 M citrate buffer pH 2.6 containing no NaCl. PMID- 10480248 TI - Partial primary structure of a fibrinogenase from the venom of the snake Lachesis stenophrys. AB - The partial primary structure of an Mr 24,000 non-haemorrhagic metalloproteinase isolated from the venom of the snake Lachesis stenophrys has been determined. The native proteinase was resistant to Edman degradation exhibiting the N-terminal blockade. The pyridylethylated or native proteinase was chemically and enzymatically fragmented and the obtained peptides were separated by gel or reversed-phase chromatography, and sequenced. The metalloproteinase from Lachesis stenophrys contains a putative zinc-chelating sequence HELGHNLGMKH, characteristic for the reprolysin family of zinc-metalloproteinases. It contains six cysteine residues in the standard positions for this group of proteins suggesting the same disulfide bonding. Interestingly, it has almost identical sequence as the metalloproteinase from Lachesis muta muta, LHF-II, which is, however, haemorrhagic. The main structural differences between the two molecules were found in their N-terminal parts and in glycosylation. As the substrate binding regions of both proteinases are practically identical, we suggest that the absence of haemorrhagicity in Lachesis stenophrys enzyme is due to its lower affinity for the matrix proteins and not due to different substrate specificity. PMID- 10480249 TI - Binding of lead to collagen type I and V and alpha2(I) CNBr (3,5) fragment by a modified Hummel-Dreyer method. AB - Binding of lead (as lead acetate) to collagen type I alpha, and alpha2 chains, collagen type V and a large cyanogen bromide fragment of type I collagen [alpha2(I)CB(3,5)] was investigated by the large-zone Hummel-Dreyer method. It was demonstrated that two categories of binding sites exist in the collagen molecule, the number of which correlates rather well with the available aspartic and glutamic acid residues. Similar results were obtained for all collagen chains (fragments) used. The number of sites thus obtained was compared with the cross striation pattern (reflecting areas where lead is bound) of the SLS form of collagen type I (alpha1 chain); it is suggested that the number of bands seen in the SLS form reflects primarily the number of available aspartic acid residues in the molecule. The association constants obtained are comparable with the low affinity interactions seen e.g., between Cu and bovine serum albumin. PMID- 10480250 TI - New polymer-based prepacked column for the reversed-phase liquid chromatographic separation of peptides over the pH range 2-12. AB - The aim of this paper was to investigate the properties of a new column, Source 5RPC, for the separation of peptides at pH 2, 4.5, 7, 9 and 12 and to compare this product with similar polymer-based products available on the market. All columns were prepacked with 5 microm polystyrene-divinylbenzene polymer bead matrices and had dimensions of 150x4.6 mm I.D. Separations of angiotensin peptides were achieved on these columns using different equilibration solvents in the pH range 2-12 and elution with acetonitrile gradients. The separation of the peptide mixture obtained on Source 5RPC column was compared with that of two other commercially available polymer-based matrices. Method scouting and optimisation were carried out using the novel chromatography system, AKTA purifier. PMID- 10480251 TI - Characterisation by immobilised metal ion affinity chromatographic procedures of the binding behaviour of several synthetic peptides designed to have high affinity for Cu(II) ions. AB - In this investigation, several peptides containing an increasing number of histidine residues have been designed and synthesised. The peptides involved repeat units of either the pentameric EAEHA or the tetrameric HLLH sequence motifs. Adsorption isotherms for these synthetic peptides and hexahistidine (hexa His) as a control substance were measured under batch equilibrium binding conditions with an immobilised Cu(II)-iminodiacetic acid (IDA) sorbent. The experimental data were analysed in terms of Langmuirean binding behaviour. In common with previous studies with synthetic peptides, these investigations have demonstrate that the sequential organisation of the histidine side chains in these peptides can affect the selectivity of the coordination interactions with borderline metal ions in immobilised metal ion affinity chromatographic systems. The results also confirm that peptides selected on the basis of their potential to form amphipathic secondary structures with their histidine residues presented on one face of the molecule can exhibit equivalent or higher affinity constants towards copper ions than hexa-His, although they contain fewer histidine residues. These findings are thus relevant to the selection of peptides produced inter alia by combinatorial synthetic procedures to have enhanced binding properties for Cu(II) or Ni(II) ions, or intended for use as peptide tags in the fusion handle approach for the affinity chromatographic purification of recombinant proteins. PMID- 10480252 TI - Application of a peptide bank from porcine brain in isolation of regulatory peptides. AB - Over the past years, the introduction of biological assay systems, random peptide sequencing and orphan receptor screening has led to the isolation and identification of new regulatory peptides with potential clinical impact. We have developed a method for separating peptides into about 300 fractions from large amounts of porcine brain tissue. The preparation of this peptide bank consists of three steps including ultrafiltration followed by cation-exchange separation and reversed-phase chromatography. These fractions represent the peptide bank with desalted and lyophilized peptides from brain tissue. Molecular masses of the peptides in the fractions are determined by matrix-assisted laser desorption ionization MS and a mass data bank is subsequently generated. For systematic analysis of the peptides, a subsequent two-step purification procedure is followed by Edman sequencing resulting in the identification of different peptides. A survival assay with a neuronal cell line revealing the stimulatory and inhibitory activities is applied as a model to test the 300 fractions. This primary screen indicates that the biological activities of the extracted peptides are easily characterized and, moreover, can be related to the biochemical entities. We conclude that the established peptide bank is an efficient and useful tool for the isolation of regulatory brain peptides applying different purification strategies. PMID- 10480253 TI - Proteolytic cleavage of glucagon-like peptide-1 by pancreatic beta cells and by fetal calf serum analyzed by mass spectrometry. AB - Fetal calf serum and a beta-cell line exhibit a proteolytic activity essential for the biological function of glucagon-like peptide-1 (GLP-1). This process of cleavage was investigated using matrix-assisted laser desorption ionization time of-flight mass spectrometry (MALDI-TOF-MS). To generate processing products, GLP 1 was subjected to rat insulinoma m5F (RINm5F) cell cultures or to fetal calf serum (FCS). For detection of processing products, a standardized extraction method including ion-exchange batch extraction, ultrafiltration, gel filtration, and reversed-phase chromatography was used. The RP fractions were analyzed by MALDI-TOF-MS. Processed proteolytic products were detected by comparing the resulting mass spectra of cell media or FCS after 2 h incubation with GLP-1 (7 36) amide with these of 2 h controls. To perform the comparison of the resulting mass spectra, software (MASSSPECANALYST) based on Microcal Software, Origins C like language LABTALK was developed. GLP-1 fragments were purified by RP-HPLC, and characterized by sequence analysis. As insulin is the major secretory product of beta cells depending on GLP-1 stimulation, the insulin and insulin fragments of the cell culture supernatants were also analyzed by this method. PMID- 10480254 TI - Separation of oligonucleotides on novel monolithic columns with ion-exchange functional surfaces. AB - Porous monolithic columns have been prepared by the direct free radical copolymerization of glycidyl methacrylate and ethylene dimethacrylate within the confines of a 50x8 mm I.D. chromatographic column in the presence of porogens. The epoxide groups of these monoliths were modified to different extents by reaction with diethylamine to afford 1-N,N-diethylamino-2-hydroxypropyl functionalities useful for ion-exchange chromatography. Following characterization of the monoliths, the columns were tested in the chromatographic separation of a homologous series of oligodeoxyadenylic [pd(A)(12-18)] and oligothymidylic acids [d(pT)(12-24)] at different flow-rates. Very good separations of the oligonucleotides were achieved even at the high flow-rate of 4 ml/min. PMID- 10480255 TI - Introduction of the gene amplification technique to decrease the risk of hepatitis C virus transmission by plasma products. AB - The viral safety of plasma-derived products with respect to hepatitis C virus (HCV) is assured by selection of donors, screening of individual donations for antibodies to HCV and the incorporation of effective viral inactivation-removal steps into manufacturing processes. As antibody screening of single donations is not sufficient to completely eliminate HCV RNA positive plasmas from plasma pools, testing for HCV RNA by gene amplification techniques may be necessary to identify positive donations. Using modern molecular biology techniques, we developed a specific, sensitive and reproducible method for routine PCR screening for HCV RNA in plasma pools. PMID- 10480256 TI - Immobilized biomembrane chromatography of highly lipophilic drugs. AB - Drug interaction with lipid bilayers was quantified by immobilized biomembrane chromatography on a series of columns containing different small amounts of human red cell membrane vesicles to extend and characterize this technique, which shows a potential for drug screening and prediction of drug absorption in humans. The chromatographic retention volume for each drug was essentially proportional to the amount of immobilized lipid, and the slope equalled the capacity factor (Ks) previously determined on single columns. Gel beds containing 0.5-2 micromol of membrane phospholipid allowed analysis of drugs with log Ks values of 2.5-4.3 in time periods of 1 min to 1 h. Highly lipophilic drugs could thus be analyzed conveniently in aqueous buffer. PMID- 10480257 TI - Experimental studies on affinity chromatography in an electric field. AB - A multicompartment electrolyzer, which has been used for preparative electrophoresis [Z. Liu, Z. Huang, J.-Y. Cong, et al., Sep. Sci. Technol. 31 (1996) 427], is applied for carrying out affinity chromatography in an alternating electric field. The effect of electric field strength on the adsorption and desorption characteristics is experimentally examined with human serum albumin and Blue Sepharose Fast Flow as a model system. It is shown that the existence of an electric field leads to a significant change in the adsorption capacity of the blue dye, which may be used for establishing a preferential adsorption to achieve a high resolution. The adsorption speed increases slightly with respect to the increase of electric field strength, while adsorption capacity in the presence of an electric field is independent of the electric field strength. Different elution behavior is observed as function of adsorption condition and a high recovery of the adsorbed protein is obtained when the adsorption is carried out in the presence of an electric field. PMID- 10480258 TI - Separation of proteins and peptides by capillary electrophoresis in acid buffers containing high concentrations of surfactants. AB - Separations of proteins at acid pH in the presence of a high concentration of surfactant [sodium laurylsulfate (SDS), 50 mmol/l] was investigated. The purpose of using high concentrations of SDS as background electrolyte modifier was threefold: First, the surfactant exerts a washing effect upon the capillary wall thus preventing binding of analytes and possible clogging of the capillary. Second, it was revealed that even under very acid conditions (below pH 3) the surfactant is capable of forming associates with protein analytes which still bear considerable negative charge and can be separated on this basis. Third, the system can be applied not only for protein mixtures sufficiently soluble in neutral to alkaline media (leukocyte lysates, standard proteins), but it can be used also with proteins, that are under such conditions virtually insoluble and their solubilization is possible in acid buffers only (eggshell proteins or collagen CNBr fragments). The result was that adsorption to the capillary wall was minimized and the analytes were separated as negatively charged associates with high efficiency. With collagen fragments partition was possible on the affinity differences of the peptides to the surfactant micelles and inner wall of the capillary. Theoretical plate counts approaching 100,000 were easily achieved even with proteins which under the more conventional operation conditions exhibit considerable sticking to the capillary wall. The other feature of this system is that the associates move very rapidly to the anode. Owing to the low pH, endoosmotic flow is negligible, and therefore the system has to be operated at reversed polarity. PMID- 10480259 TI - Capillary electrophoretic study of cisplatin interaction with nucleoside monophosphates, di- and trinucleotides. AB - cis-Diamminedichloroplatinum(II) (cisplatin) is applied against different kinds of cancer although toxic side effects are known. Screening systems for alternative compounds with higher effectiveness but minimizing toxic side effects are required. We investigated the adduct formation of cisplatin with nucleoside monophosphates, di- and trinucleotides. Capillary electrophoretic separations were performed in a sodium phosphate buffer using an instrument equipped with a diode array detector. Adduct formation results in a significant shift of lambda(max) to lower energy compared to free nucleotides. Therefore, UV spectra are an important tool for peak identification. We could separate and identify all four common nucleotides and their major platinum adducts in a single run demonstrating the suitability of CE for these kinds of investigations. Furthermore, kinetic studies of these reactions are performed. PMID- 10480260 TI - Antiviral drug susceptibility assays: going with the flow. AB - This review describes the procedures for the use of fluorochrome labeled monoclonal antibodies and flow cytometry for the detection and quantification of virus infected cells. The application of this technology for (1) identifying virus infected cells in clinical specimens obtained from human cytomegalovirus (HCMV) and human immunodeficiency virus (HIV) infected individuals; (2) screening antiviral compounds active against HCMV, HDSV and HIV; and (3) performing drug susceptibility testing for HCMV, HSV and HIV clinical isolates are reviewed. The flow cytometry drug susceptibility assay is rapid, quantitative, and easily performed. It should be considered by anyone interested in performing drug susceptibility testing for any virus for which there are reliable monoclonal antibodies. PMID- 10480261 TI - Selective activity of various antiviral compounds against HHV-7 infection. AB - Human herpesvirus virus type 7 (HHV-7) is a T-lymphotropic herpesvirus which uses the CD4 receptor as main receptor to infect its target cells. Measuring the decrease of CD4 expression during HHV-7 infection is a convenient and accurate method to monitor the efficacy of antiviral agents against HHV-7 infection. Different classes of compounds, such as heparin, pentosan polysulfate (PS), dextran sulfate (DS), aurintricarboxylic acid (ATA), phosphonoformic acid (PFA), 9-(2-phosphonylmethoxyethyl)adenine (PMEA), 2-amino-7-[(1,3-dihydroxy-2-propoxy) methyl] purine (S2242), polyvinylalcohol sulfate (PVAS) and the co-polymer of vinylalcohol sulfate with acrylic acid (PAVAS), acyclovir (ACV), ganciclovir (GCV), penciclovir (PCV), brivudin (BVDU), cidofovir (HPMPC), lobucavir, (R)-9-[4 hydroxy-2-(hydroxymethyl)butyl]guanine] (H2G), (R)-9-(2 phosphonylmethoxypropyl)adenine (PMPA) and sorivudine (BVaraU), were evaluated for their anti-HHV-7 activity in the SupT1 T cell line and in purified CD4+ T lymphocytes. Antiviral activity was monitored by inhibition of: (i) CD4 expression down-regulation; (ii) giant cell formation and (iii) apoptosis induction. In general, PS, DS, PVAS, PAVAS, ATA, PFA, PMEA, S2242, lobucavir and HPMPC had comparable anti-HHV-7 activity in the two cell lines, irrespective of the parameters followed to monitor antiviral activity. One of the exceptions was heparin which had an IC50 of 9.6 microg/ml in SupT1 cells and >250 microg/ml in CD4+ T lymphocytes. The compounds PCV, GCV, H2G and PMPA showed some activity in CD4+ T lymphocytes, but not in SupT1 cells. ACV, BVDU and BVaraU did not show activity in either cell system. None of the chemokines tested, such as platelet factor-4 (PF-4), eotaxin, stromal cell-derived factor 1alpha(SDF-1alpha) and RANTES, had detectable activity against HHV-7. In contrast, the HIV-1 envelope glycoprotein, gp120, and the two anti-CD4 mAbs, 13B8-2 and OKT4, were clearly active against HHV-7 infection. PMID- 10480262 TI - Synthesis and antiviral activity of phosphoralaninate derivatives of methylenecyclopropane analogues of nucleosides. AB - Phenylmethylphosphoro-L-alaninate prodrugs of antiviral Z-methylenecyclopropane nucleoside analogues and their inactive E-isomers were synthesized and evaluated for their antiviral activity against HCMV, HSV-1, HSV-2, HHV-6, EBV, VZV, HIV-1 and HBV. The adenine Z-analogue was a potent inhibitor of all these viruses but it displayed cellular toxicity. The guanine Z-derivative was active against HCMV, HBV, EBV and VZV and it was not cytotoxic. The 2,6-diaminopurine analogue was the most potent against HIV-1 and HBV and somewhat less against HHV-6, HCMV, EBV and VZV in a non-cytotoxic concentration range. The 2-amino-6-cyclopropylamino and 2 amino-6-methoxypurine prodrugs were also more active than parent analogues against several viruses but with a less favorable cytotoxicity profile. In the E series of analogues, adenine derivative was active against HIV-1, HBV and EBV, and it was non-cytotoxic. The guanine analogue exhibited a significant effect only against HBV. The 2,6-diaminopurine E-analogue was inactive with the exception of a single EBV assay. The 2-amino-6-methoxypurine Z methylenecyclopropane nucleoside analogue was an effective inhibitor of HCMV, MCMV and EBV. The 2,6-diaminopurine Z-prodrug seems to be the best candidate for further development. PMID- 10480264 TI - In situ imaging of microorganisms in geologic material. AB - In order to fully delineate the interactions of microorganisms with geological substrates, unequivocal identification of intact microbial cells within geologic samples is required without the disruption of either the rock texture or the relationship of the microorganisms to the mineral fabric. To achieve this objective we developed a protocol that enables the visualization of intact microbial cells in petrographic thin sections, avoids detaching the cells from their host mineral surfaces and avoids microbial contamination during the lapidary process. Propidium iodide and POPO-3, nucleic acid stains that specifically target double-stranded DNA and RNA were utilized for in situ visualization of cells in surface and subsurface basalts from northeastern Idaho. Additionally, examination of samples incubated with acetic acid-UL-14C via phosphor imagining facilitated the in situ visualization of 14C labeled biomass. Biomass observed was low (<10(7) cells/g). These observations indicate that the microbial distribution in these rocks exhibits a high degree of spatial heterogeneity at the sub-centimeter scale. PMID- 10480263 TI - Enhancement of the innate and cellular immune response in patients with genital warts treated with topical imiquimod cream 5%. AB - The mechanism of action of imiquimod 5% cream applied topically to patients with genital warts was evaluated in a double-blind, placebo-controlled study. Imiquimod (16 patients) or placebo (three patients) was applied three times per week for up to 16 weeks. All imiquimod-treated patients had a > or =75% reduction in total wart area while only one of three placebo-treated patients had a similar reduction. Wart biopsies were taken at prestudy, week 6, and end of treatment. Polymerase chain reaction (PCR) for human papillomavirus (HPV) DNA and reverse transcriptase (RT)-PCR for messenger (m)RNAs were used to identify cytokines, cellular markers, viral gene products, and cell cycle markers in these biopsies. Treatment with imiquimod, an immune response modifier, stimulated significant increases in mRNA for interferon (IFN)-alpha, IFN-gamma and 2',5' oligoadenylate synthetase (2',5'-AS) as well as a tendency towards increases in tumor necrosis factor (TNF)-alpha and interleukin-12 p40. Significant increases in mRNA for CD4 and a trend toward increases in CD8 were also observed in imiquimod-treated patients, suggesting activation of a cell mediated immune response. Imiquimod administration was also associated with a significant decrease in viral load as measured by HPV DNA and L1 mRNA. The effects on HPV markers were accompanied by an apparent decrease in mRNA expression for markers of cell proliferation and an increase in mRNA for markers of keratinocyte differentiation and tumor suppressors. PMID- 10480265 TI - Development and validation of an automated, microscopy-based method for enumeration of groups of intestinal bacteria. AB - An automated microscopy-based method using fluorescently labelled 16S rRNA targeted oligonucleotide probes directed against the predominant groups of intestinal bacteria was developed and validated. The method makes use of the Leica 600HR image analysis system, a Kodak MegaPlus camera model 1.4 and a servo controlled Leica DM/RXA ultra-violet microscope. Software for automated image acquisition and analysis was developed and tested. The performance of the method was validated using a set of four fluorescent oligonucleotide probes: a universal probe for the detection of all bacterial species, one probe specific for Bifidobacterium spp., a digenus-probe specific for Bacteroides spp. and Prevotella spp. and a trigenus-probe specific for Ruminococcus spp., Clostridium spp. and Eubacterium spp. A nucleic acid stain, 4',6-diamidino-2-phenylindole (DAPI), was also included in the validation. In order to quantify the assay error, one faecal sample was measured 20 times using each separate probe. Thereafter faecal samples of 20 different volunteers were measured following the same procedure in order to quantify the error due to individual-related differences in gut flora composition. It was concluded that the combination of automated microscopy and fluorescent whole-cell hybridisation enables distinction in gut flora-composition between volunteers at a significant level. With this method it is possible to process 48 faecal samples overnight, with coefficients of variation ranging from 0.07 to 0.30. PMID- 10480266 TI - Use of PCR-restriction fragment length polymorphism analysis of the hsp65 gene for rapid identification of mycobacteria in Brazil. AB - Polymerase chain reaction amplification of part of the gene coding for the heat shock protein hsp65 followed by restriction enzyme analysis (PRA) is a recently described tool for rapid identification of mycobacteria. In this study, the speed and simplicity of PRA for identification of isolates of mycobacteria from patients with clinical symptoms of tuberculosis was evaluated and compared with identification results obtained by commercially available methods. Established PRA patterns were observed for nineteen isolates of Mycobacterium tuberculosis, eleven belonging to the complex M. avium-intracellulare, four of M. kansasii, one of M. fortuitum, one of M. abscessus, three of M. gordonae and one of the recently described species M. lentiflavum, as identified by commercially available methods. Two isolates of M. fortuitum and one of M. gordonae had unique and so far undescribed PRA patterns, suggesting geographically-related intra species variation within the hsp65 sequence. We propose the inclusion of these new patterns in the PRA identification algorithm and have defined more accurately the molecular weight values of the restriction fragments. This is the first report on the isolation of M. lentiflavum in Brazil suggesting that identification by means of PRA could be useful for detection of mycobacterial species that are usually unnoticed. Where the use of several commercial techniques in combination was necessary for correct identification, PRA demonstrated to be a simple technique with good cost-benefit for characterization of all mycobacterial isolates in this study. PMID- 10480267 TI - Application of a tetrazolium dye as an indicator of viability in anaerobic bacteria. AB - The use of the redox dye 5-cyano-2,3,-ditolyl tetrazolium chloride (CTC) for evaluating the metabolic activity of aerobic bacteria has gained wide application in recent years. In this study, we examined the utility of CTC in capturing the metabolic activity of anaerobic bacteria. In addition, the factors contributing to abiotic reduction of CTC were also examined. CTC was used in conjunction with the fluorochrome 5-(4,6-dichlorotriazinyl) aminofluorescein (DTAF), that targets bacterial cell wall proteins, to quantitate the active fraction of total bacterial numbers. Facultative anaerobic bacteria, including Escherichia coli grown fermentatively, and Pseudomonas chlorophis, P. fluorescens, P. stutzeri, and P. pseudoalcalegenes subsp. pseudoalcalegenes grown under nitrate-reducing conditions, actively reduced CTC during all phases of growth. Greater than 95% of these cells accumulated intracellular CTC-formazan crystals during the exponential phase. Obligate anaerobic bacteria, including Syntrophus aciditrophicus grown fermentatively, Geobacter sulfurreducens grown with fumarate as the electron acceptor, Desulfovibrio desulfuricans subsp. desulfuricans and D. halophilus grown under sulfate-reducing conditions, Methanobacterium formicicum grown on formate, H2 and CO2, and Methanobacterium thermoautotrophicum grown autotrophically on H2 and CO2 all reduced CTC to intracellular CTC-formazan crystals. The optimal CTC concentration for all organisms examined was 5 mM. Anaerobic CTC incubations were not required for quantification of anaerobically grown cells. CTC-formazan production by all cultures examined was proportional to biomass production, and CTC reduction was observed even in the absence of added nutrients. CTC was reduced by culture fluids containing ferric citrate as electron acceptor following growth of either G. metallireducens or G. sulfurreducens. Abiotic reduction of CTC was observed in the presence of ascorbic acid, cysteine hydrochloride, dithiothreitol, NADH, NADPH, Fe(II)Cl2, sodium thioglycolic acid and sodium sulfide. These results suggest that while CTC can be used to capture the metabolic activity of anaerobic bacteria, care must be taken to avoid abiotic reduction of CTC. PMID- 10480268 TI - A PCR test for the identification and discrimination of Legionella longbeachae serogroups 1 and 2. AB - A PCR test has been developed for the specific identification of Legionella longbeachae. The test targeted sequence unique to both L. longbeachae serogroups 1 and 2 within the mip gene and permitted both species and serogroup identification. The test was trialed on a range of closely related species and 20 clinical isolates originating from Australia, the USA and Israel. Results were consistent with previous identification analyses. From 20 water samples known to contain Legionella spp. one sample yielded isolates which consistently tested positive by L. longbeachae serogroup 1 PCR. DNA sequencing of the PCR product, 5S rRNA gene sequence and hybridisation analysis with a specific oligonucleotide probe definitively identified one isolate as L. longbeachae serogroup 1. PCR testing was demonstrated as a superior method of identification to traditional seroagglutination reactions, which were ambiguous and could explain the previous failure to identify the presence of this microorganism in water. PMID- 10480269 TI - Determining chemotactic responses by two subsurface microaerophiles using a simplified capillary assay method. AB - A simplified capillary chemotaxis assay utilizing a hypodermic needle, syringe, and disposable pipette tip was developed to measure bacterial tactic responses. The method was applied to two strains of subsurface microaerophilic bacteria. This method was more convenient than the Adler method and required less practice. Isolate VT10 was a strain of Pseudomonas syringae, which was isolated from the shallow subsurface. It was chemotactically attracted toward dextrose, glycerol, and phenol, which could be used as sole carbon sources, and toward maltose, which could not be used. Isolate MR100 was phylogenetically related to Pseudomonas mendocina and was isolated from the deep subsurface. It showed no tactic response to these compounds, although, it could use dextrose, maltose, and glycerol as carbon sources. The chemotaxis results obtained by the new method were verified by using the swarm plate assay technique. The simplified technique may be useful for routine chemotactic testing. PMID- 10480270 TI - Facilitation of neuronal activity in somatosensory and posterior parietal cortex during prehension. AB - In order to study prehension in a reproducible manner, we trained monkeys to perform a task in which rectangular, spherical, and cylindrical objects were grasped, lifted, held, and lowered in response to visual cues. The animal's hand movements were monitored using digital video, together with simultaneously recorded spike trains of neurons in primary somatosensory cortex (S-I) and posterior parietal cortex (PPC). Statistically significant task-related modulation of activity occurred in 78% of neurons tested in the hand area; twice as many cells were facilitated during object acquisition as were depressed. Cortical neurons receiving inputs from tactile receptors in glabrous skin of the fingers and palm, hairy skin of the hand dorsum, or deep receptors in muscles and joints of the hand modulated their firing rates during prehension in consistent and reproducible patterns. Spike trains of individual neurons differed in duration and amplitude of firing, the particular hand behavior(s) monitored, and their sensitivity to the shape of the grasped object. Neurons were classified by statistical analysis into groups whose spike trains were tuned to single task stages, spanned two successive stages, or were multiaction. The classes were not uniformly distributed in specific cytoarchitectonic fields, nor among particular somatosensory modalities. Sequential deformation of parts of the hand as the task progressed was reflected in successive responses of different members of this population. The earliest activity occurred in PPC, where 28% of neurons increased firing prior to hand contact with objects; such neurons may participate in anticipatory motor control programs. Activity shifted rostrally to S-I as the hand contacted the object and manipulated it. The shape of the grasped object had the strongest influence on PPC cells. The results suggest that parietal neurons monitor hand actions during prehension, as well as the physical properties of the grasped object, by shifting activity between populations responsive to hand shaping, grasping, and manipulatory behaviors. PMID- 10480271 TI - Motion-responsive regions of the human brain. AB - Functional magnetic resonance imaging was used to map motion responsive regions of the human brain by contrasting passive viewing of moving and stationary randomly textured patterns. Regions were retained as motion responsive if they reached significance either in the group analysis or in the majority of hemispheres in single-subject analysis. They include well-known regions, such as V1, hMT/V5+, and hV3A, but also several occipito-temporal, occipito-parietal, parietal, and frontal regions. The time course of the activation was similar in most of these regions. Motion responses were nearly identical for binocular and monocular presentations. Flicker-induced-activation introduced a dichotomy amongst these motion responsive regions. Early occipital and occipito-temporal regions responded well to flicker, while flicker responses gradually vanished as one moved to occipito-parietal and then parietal regions. Finally, over a more than four-fold range, stimulus diameter had little effect on the motion activations, except in V1. PMID- 10480272 TI - Spatiotemporal reorganization of electrical activity in the human brain associated with a timing transition in rhythmic auditory-motor coordination. AB - We used a 61-channel electrode array to investigate the spatiotemporal dynamics of electroencephalographic (EEG) activity related to behavioral transitions in rhythmic sensorimotor coordination. Subjects were instructed to maintain a 1:1 relationship between repeated right index finger flexion and a series of periodically delivered tones (metronome) in a syncopated (anti-phase) fashion. Systematic increases in stimulus presentation rate are known to induce a spontaneous switch in behavior from syncopation to synchronization (in-phase coordination). We show that this transition is accompanied by a large-scale reorganization of cortical activity manifested in the spatial distributions of EEG power at the coordination frequency. Significant decreases in power were observed at electrode locations over left central and anterior parietal areas, most likely reflecting reduced activation of left primary sensorimotor cortex. A second condition in which subjects were instructed to synchronize with the metronome controlled for the effects of movement frequency, since synchronization is known to remain stable across a wide range of frequencies. Different, smaller spatial differences were observed between topographic patterns associated with synchronization at low versus high stimulus rates. Our results demonstrate qualitative changes in the spatial dynamics of human brain electrical activity associated with a transition in the timing of sensorimotor coordination and suggest that maintenance of a more difficult anti-phase timing relation is associated with greater activation of primary sensorimotor areas. PMID- 10480273 TI - Comparison of input-output patterns in the corticospinal system of normal subjects and incomplete spinal cord injured patients. AB - We have examined input-output patterns in the corticospinal system after incomplete spinal cord injury. The amplitude of the motor evoked potential (MEP) to transcranial magnetic stimulation (TMS) was used to study the patterns of recruitment, with increasing stimulus intensity, and facilitation, with increasing voluntary contraction, in thenar muscles of 12 patients with incomplete spinal cord injuries and 13 control subjects. The patients had all suffered spinal cord injury at a segmental level rostral to C8 and T1, the segments supplying innervation of thenar muscles. The patients showed a less pronounced increase in MEP amplitude with increasing strength of TMS compared with the controls. Specifically, at a stimulus strength of 120% threshold and above, the patients showed significantly smaller MEPs relative to the maximum ulnar nerve M-wave response than the controls. The patients also showed a less steep pattern of facilitation with voluntary drive. The MEP continued to increase up to 50% maximum voluntary contraction (MVC) whereas the controls reached a plateau around 10% MVC. The results indicate that the patients show modified corticospinal recruitment and facilitation of the motoneurone pool. We speculate that the function of the adapted corticospinal system after spinal cord injury might be to regulate and modulate drive to motoneurones originating from segmental and other descending inputs. We discuss how such a modified corticospinal system might be of functional benefit to the patients. PMID- 10480274 TI - A possible selective impairment of magnocellular function in compression of the anterior visual pathways. AB - Two parallel visual systems, the magnocellular (M) and parvocellular (P) pathways, originate from different types of retinal ganglion cells, and are known to be segregated in different portions of the pregeniculate visual pathways. Their relative contribution to two main cortical streams, dorsal and ventral, is still under discussion, but it is reasonable to suppose that selective damage to the M or P subcortical system might interfere with specific aspects of processing within one or the other cortical system. Using two different apparent-motion tasks, we compared the performance of patients affected by compression of the ventral part of the pregeniculate visual pathways with that of normal controls. In the first task, observers detected small displacements of a low-contrast vertical bar, while in the second task they estimated the visible persistence of moving dots. In the first task, patients were impaired with parafoveal displays, especially in the temporal portion of the visual field. In the second task, patients showed reduced suppression of visible persistence at long, but not at short, exposure durations. Three considerations support the hypothesis that these results represent a selective impairment of the M system. First, M axons are more likely to suffer from compression, particularly in the case of a mass growing from below since they are known to occupy a ventral subpial position in the optic chiasm and tract. Second, the performance of patients with a ventral compression is consistent with the characteristics of the response properties of P ganglion cells, which have previously been shown to exhibit elevated and unmodulated thresholds for displacement detection in the macaque monkey. Third, such patients are less sensitive to the inhibitory signals that suppress visible persistence, which probably originate in the M system. PMID- 10480275 TI - Specific and non-specific effects of transcranial magnetic stimulation on simple and go/no-go reaction time. AB - The effects of subthreshold transcranial magnetic stimulation (TMS) on simple and go/no-go reaction time (RT) tasks were studied in seven healthy volunteers. Subjects were asked to respond by abducting the thumb in a warning-imperative signal paradigm. TMS was randomly delivered at variable delays to the imperative signal (IS). Simple RT was significantly shortened when TMS was delivered to the left motor cortex and parietal regions simultaneously with IS. In the go/no-go paradigm, a similar trend to shorter RT was seen at a delay of 0 ms. Additionally, a significant shortening was observed at a delay of 90 ms with TMS over the contralateral motor cortex only. Movement-related potentials (MRPs) in the two paradigms showed a predominantly contralateral negativity approximately 80 ms preceding EMG onset. Our findings support the existence of two differentiated effects of TMS on RT: (1) one non-specific effect, evidenced in both the simple and go/no-go paradigms at a 0 ms delay, which can be at least partially explained by intersensory facilitation; and (2) a motor-specific effect of TMS, unveiled in the go/no-go paradigm at a 90 ms delay. PMID- 10480276 TI - Consequences of altered cerebellar input for the cortical regulation of motor coordination, as reflected in EEG potentials. AB - The cerebellum is certainly involved in fine coordination of movements, but has no efferences of its own to the muscles. Thus, it can exert its influence only via other cerebral areas that have those efferences. This study investigated in patients with cerebellar atrophy how cortical motor areas are affected by dysfunction of the cerebellum. The main question was whether the patients' slow cortical electroencephalogram (EEG) potentials during key-press preparation and execution would be generally altered or would be specifically altered when fine coordination was needed. In the coordination task, right- and left-hand keys had to be pressed simultaneously with different forces, under visual feedback. Control tasks were to press with both hands equally or with one hand only. The patients indeed had a performance deficit in the coordination task. Their cortical EEG potentials were already drastically reduced in the simple tasks, but were enhanced by the same amount as in healthy subjects when more coordination was needed. These results suggest that the cerebellum is not exclusively active in fine coordination, but is generally involved in any kind of preparatory and executive activity, whereas the motor cortex becomes more active with fine coordination. The role of the cerebellum might be to provide the motor cortex with information needed for coordinating movements. In cerebellar atrophy, this altered input may be sufficient for the motor cortex in controlling simple tasks, but not for complex ones. PMID- 10480277 TI - Locus of control beliefs predicting oral and diabetes health behavior and health status. AB - Our study evaluates the correlation between dental and diabetes locus of control beliefs and the potentiality of locus of control beliefs in predicting oral health behavior, dental status, diabetes compliance, and HbA1c level by using situation-specific locus of control scales and considering the value dimension. Data were collected by means of a quantitative questionnaire, a clinical oral examination and patient records. The research population comprised 149 insulin dependent diabetics who had teeth of their own. Variables were the frequencies of tooth brushing and dental visiting, oral indexes, diabetes adherence, and HbA1c level. Dental and diabetes locus of control beliefs correlated with each other. Dental locus of control associated with frequency of dental visiting, plaque index, decayed surfaces, and with root caries, but diabetes locus of control associated only weakly with adherence with diabetes self-care regimens and not at all with HbA1c level. Correlations between dental locus of control and oral indexes were stronger among those having high value for dental care. Although there were correlations between dental and diabetes locus of control beliefs, only dental locus of control beliefs are practicable for determining health behavior and health status. It is therefore concluded that locus of control beliefs are health behavior specific. PMID- 10480278 TI - Explanatory models for clinically determined and symptom-reported caries indicators in an adult population. AB - The aim of the present study was to analyze possible indicators of: (i) relative number of decayed and filled teeth, (ii) relative number of decayed teeth, (iii) subjectively reported toothache, and (iv) sensitive teeth, and to find explanatory models for these phenomena. Independent variables from three domains were used: (i) socio-economic factors, (ii) general health and health-related lifestyle, and (iii) dental attitudes and behaviors. The study basis was validated questionnaires from all 50-year-olds in 2 Swedish counties (n = 8888), response rate 71% (n = 6343). For a 20% subsample (58% participation) the DFT and DT were determined by calibrated dentists. Analyses were done with logistic and multiple regression. The variables born outside Sweden, gender, education, shift work, satisfaction with dental care, fear and care utilization were associated with DFT/number of teeth. For DT/number of teeth, the direction of association was reversed for the variables born outside Sweden and gender. Social class, education, general health, and use of tobacco were further covariates. Good oral hygiene gave a lower ratio of DT. For the logistic regression model of toothache, residence in cities and satisfaction with dental care had lower probability for toothache reports, while born outside Sweden, mouth dryness, use of pharmaceuticals, tobacco, fear, and high utilization increased this probability. In general, the association pattern was as could be expected: immigrants, working class, low education, smoking, dissatisfaction with dental treatment and low utilization all appeared as risk factors for both the clinically determined caries indicators, but not necessarily for subjective symptom reports. Only fear of dental treatment showed a consistent positive association with all the indicators. PMID- 10480279 TI - The perceptions of users about barriers to the use of free systematic oral care among Finnish pre-school children--a qualitative study. AB - With the economic recession in the early 1990s in mind, and the change in the provision of healthcare services, our aim was to explore qualitatively what barriers families of Finnish pre-school children had perceived related to the use of free systematic oral healthcare. Qualitative interviews were carried out with the parents of 12 pre-school children attending an oral health examination to ascertain what barriers those using oral services had overcome, and what they considered to be the reasons for the non-attendance of other families. Content analysis was used by classifying groups of barriers and their reported importance as well as the reasons for non-attendance. The barriers discovered were related to: difficulties in the (i) change of daily routines or (ii) booking time for care, (iii) poor coordination with other healthcare services, (iv) fear and negative image of oral care, and (v) inconsistent content of oral health information. When parents replied to the externalized question about reasons for the non-attendance of some families, they most often mentioned difficulties in changing daily routines, laziness, lack of interest and fear. Barriers to attending oral healthcare could be lowered by emphasizing the positive image of oral health services, by providing more effective coordination with mother and child health services, and by providing appointment times later in the day. PMID- 10480280 TI - Interdental caries incidence and progression in relation to mutans streptococci suppression after chlorhexidine-thymol varnish treatments in schoolchildren. AB - The aim of this study was to evaluate interdental caries incidence and progression in relation to the effect of antibacterial varnish treatments in schoolchildren anticipated at caries risk. After a screening procedure, 110 healthy children (8-10 years) with moderate and high counts of salivary mutans streptococci (MS) were invited to join a 2-year longitudinal study. At baseline, MS were enumerated at all mesial interdental sites of the first permanent molars with a chair-side technique. The children were then treated 3 times within 2 weeks by interdental spot applications with a 1% chlorhexidine/thymol-containing varnish (Cervitec). Follow-up samples of saliva and plaque were collected 1, 3, 6, and 12 months after onset of treat. Caries prevalence, incidence and progression of the selected approximal surfaces were scored from bitewing radiographs exposed at baseline and after 2 years. Sixty-three children of the same age formed a non-varnish-treated reference group. Reduction of caries incidence and progression was dearly dependent on the outcome of the antibacterial treatment. A significantly (P< 0.01) higher progression score was found among children who exhibited a less marked suppression of interdental MS levels compared with those with high suppression and the children of the reference group. The results suggest that a suppression of MS in interdental plaque may be an important event to prevent and arrest approximal caries development in schoolchildren at risk. Monitoring the effect of antibacterial agents in a site-specific way could therefore be advocated. PMID- 10480281 TI - Caries assessment and restorative treatment thresholds reported by Swedish dentists. AB - The aim was to study any variability in approximal and occlusal caries diagnoses and restorative treatment decisions among Swedish dentists. The material consisted of a pre-coded questionnaire sent to a random sample of 923 dentists with 4 items concerning approximal and occlusal caries diagnosis and restorative treatment decisions. Responses were received from 651 (70.5%) dentists. In an adolescent with low caries activity and good oral hygiene, more than 90% of the dentists stated that they would not automatically restore a primary approximal caries lesion if its radiographic appearance did not show obvious progression in the outer 1/3 to 1/2 of the dentin. Moreover, 67% of the dentists would only consider immediate restorative treatment of an occlusal surface if obvious cavitation and/or radiographic signs of dentin caries could be observed. When diagnosing questionable occlusal caries, the dentists largely relied on the radiographic appearance. Concerning both approximal and occlusal caries, the threshold for restorative treatment differed between the metropolitan regions in Sweden, and younger more often than older dentists would postpone restorative treatment of approximal caries until the lesion had reached a relatively advanced stage of progression. The responses also showed that dentists in private practice would restore approximal caries at an earlier stage of progression than the dentists in the Public Dental Health Service. PMID- 10480282 TI - Cervical thickness of the mandibular alveolar process and skeletal bone mineral density. AB - Much effort has been devoted to finding methods for detecting individuals with low bone mass and risk of osteoporotic fractures. The aim of the present study was to investigate whether there is a relationship between the thickness of the alveolar process and the bone mineral density (BMD) of the distal forearm. In 24 women (38-65 years), the BMD of the distal forearm, obtained by dual X-ray absorptiometry, was correlated to the difference between two measures of the thickness of the mandibular alveolar process in the region of the first premolar. A highly significant correlation (r = 0.95, P< 0.001) was found. The method was cross-validated by using the equation obtained from the linear regression analysis above to predict BMD in two other groups. In both groups, the correlation between the measured BMD of the forearm and the predicted BMD was highly significant (r = 0.91, P< 0.001). The interdental thickness between the canine and the second incisor was also correlated to BMD, but with lower predictive value (r = 0.67, P<0.001). Measurements of the mandibular alveolar process can be used as one of several parameters to predict skeletal bone density. PMID- 10480283 TI - Dental status of women in a 24-year longitudinal and cross-sectional study. Results from a population study of women in Goteborg. AB - The aim of the study was to describe dental health status among middle-aged and elderly women over a 24-year period. Because of the design of the study it was possible to make both longitudinal and cross-sectional comparisons. The study started in 1968-69 with a combined medical and dental examination of women aged between 38 and 60 years. New dental examinations of these same women were performed in 1980-81 and again in 1992-93, and included new cohorts of 38-year old women on both occasions. In the cross-sectional perspective, it was shown that the number of edentate individuals decreased significantly during the 24 year period. Among dentate women, the number of remaining teeth and restored teeth increased significantly cross-sectionally. However, the youngest age group, women of 38 years, showed a lower number of restored teeth in the latest study (in 1992-93). There was also a lower number of crowns, root-fillings, and pontics in the latest study for the youngest age group. The two older age groups studied cross-sectionally showed similar numbers in all studies. In the longitudinal study, there was a decrease with time in incidence of edentulism. Among the dentate women in the longitudinal study the number of restored teeth related to those remaining was high (range 76-90%) and did not change much between the studies in the different age groups. There was a clear tendency during the 24 year period in all age groups for more teeth to be restored with crowns rather than different fillings. In conclusion, this population study, with a follow-up of 24 years, shows that dental status improved in that fewer individuals lost all their teeth and younger age groups have more remaining teeth and fewer restorations than previously. PMID- 10480284 TI - Serum mercury concentration in relation to survival, symptoms, and diseases: results from the prospective population study of women in Gothenburg, Sweden. AB - A prospective population study of women in Gothenburg, Sweden was started in 1968 69 and comprised 1462 women aged 38, 46, 50, 54, or 60 years at baseline. Follow up studies were carried out in 1974-75, 1980-81, and 1992-93. The baseline study included an extensive medical and dental examination. Serum mercury concentration (beta-HG) was determined in deep-frozen samples from all participants in 1968-69 and in a random subsample of sera from participants in 1980-81, about 20 years after the baseline examination. S-Hg was statistically significantly correlated with number of amalgam fillings at both examinations. Of 30 defined symptoms and 4 different clusters of symptoms, no one was independently correlated with S-Hg measured in the samples from 1968-69, while there was a negative statistically significant correlation with over-exertion and poor appetite in 1980-81. Blood hemoglobin and serum B-12 concentrations in 1968-69 were statistically significantly and positively correlated with S-Hg, while erythrocyte sedimentation rate and the serum concentrations of potassium and triglycerides were significantly and negatively correlated with S-Hg, also after including potential confounders. Blood hematocrit examined in 1980-81 was negatively correlated with S-Hg. When including potential confounders, serum IgA was also statistically significantly correlated with S-Hg, but not in univariate analysis. No statistically significant correlation was observed between S-Hg, on the one hand, and the incidence of diabetes, myocardial infarction, stroke, or cancer on the other, while a statistically significant negative correlation was observed with overall mortality when age and education were included as background variables. There were some correlations between biological variables and S-Hg, probably of no negative clinical significance, and we conclude that there is no association between disease and S-Hg on a population basis in middle-aged and older women. PMID- 10480285 TI - Effect of recombinant human erythropoietin in preterm infants. AB - Twenty-five premature infants (mean gestational age+/-SD, 31.4+/-1.9 weeks) were administered subcutaneously recombinant human erythropoietin (rHuEpo) at a dose of 300 u/kg of body weight three times a week beginning on the third day of life and continuing for 6 weeks. The controls (n=23) were premature infants with a mean gestational age of 32.2+/-2.3 weeks who did not receive rHuEpo. Haematological indices, haemoglobin and serum phosphate (Pi), and red blood cell (RBC) phosphate metabolites (ATP, 2,3-DPG, RBCPi) were tested monthly until the 6th month and thereafter at the 9th and 12th months of life. The level of serum soluble transferrin receptors (sTfR) correlated significantly with rHuEpo (p<0.05). The ratio of sTfR to log (ferritin) was significantly higher (p<0.001) in the infants treated with rHuEpo than the controls. Intracellular organic and inorganic Pi changes were not affected by the Epo administration. The RBC 2,3-DPG seemed adequate in infants receiving rHuEpo. PMID- 10480286 TI - Retrospective study of candidemia in patients with hematological malignancies. Clinical features, risk factors and outcome of 76 episodes. AB - A retrospective study of 76 episodes of candidemia in 73 patients with underlying hematological malignancy, from 1988 until 1997, has been conducted to evaluate the clinical characteristics and to ascertain the variables related to the onset and the outcome of candidemia. The most frequent malignancy was acute myeloid leukemia (29 episodes). Candidemia developed mainly during aplasia in patients refractory to chemotherapy (42%). In 65 episodes (86%) the patients were neutropenic (ANC <1 x 10(9)/l) before the candidemia diagnosis for a median time of 13 d, and in 53 episodes (70%) at microbiological diagnosis of candidemia ANC was <1 x 10(9)/l. Candida albicans was the most frequently isolated etiologic agent (31 episodes), but C. non-albicans species sustained the majority of candidemia. Seventeen candidemias developed during azoles prophylaxis. One month after the diagnosis of candidemia, 26 patients died. In 19 cases, death was attributable to candidemia. The case-control study demonstrated, at univariate analysis, that the colonization with Candida. spp. (p=0.004), antimycotic prophylaxis (p=0.01), presence of central venous catheter (p=0.01), neutropenia (p=0.002), and the use of glycopeptide (p=0.0001) increased the risk of candidemia. Using multivariate regression analysis only colonization with Candida spp. and the previous therapy with glycopeptide were associated with a significantly increased risk. Acute mortality, expressed by a cumulative probability of survival at 30 d from diagnosis of candidemia, was 0.67 (95% C.I. 0.55-0.77) and was significantly reduced in patients with neutrophils <1 x 10(9)/l when compared to those with neutrophils >1 x 10(9)/l (p at Mantel Cox=0.029). Overall cumulative probability of survival at 1 yr was 0.38 (95% C.I. 0.27-0.49) and only the treatment with Amfotericin B significantly reduced the risk of death. PMID- 10480287 TI - Wilms' tumour gene (wt1) expression at diagnosis has no prognostic relevance in childhood acute lymphoblastic leukaemia treated by an intensive chemotherapy protocol. AB - Expression of the Wilms' tumour gene (wt1) has been demonstrated in a large proportion of human acute leukaemias and is thought to play a role in leukaemogenesis. Recent observations in adult patients with acute leukaemia suggest that wt1 gene expression is a poor prognostic factor. In childhood acute leukaemia, the clinical role of wt1 gene expression has not been established. We have therefore investigated bone marrow samples from 50 children with acute lymphocytic leukaemia at the time of diagnosis for the presence of wt1 transcripts to determine whether wt1 gene expression is associated with specific characteristics of leukaemic cells and whether it is predictive of response to treatment. All patients were treated according to the ALL-BFM 90 protocol. The median observation time was 30 months. Wt1 transcripts were detected by RT-PCR in 60% of the diagnostic samples. Wt1 PCR positive patients showed a higher median leukocyte and peripheral blast cell count than wt1 negative patients. High and intermediate risk patients more frequently displayed wt1 transcripts than low risk patients. No correlation between wtl gene expression and FAB type, immunophenotype, co-expression of myeloid antigens or karyotype has been observed. Furthermore, there was no correlation between wt1 gene expression at diagnosis and achievement of complete remission (CR) and no difference in disease free survival (DFS) or overall survival (OS) between wt1 positive and negative patients (p > 0.1). These data indicate that (1) wt1 gene expression at diagnosis is detected more frequently in patients with high leukocyte and peripheral blast cell counts, but is not associated with specific characteristics of leukaemic cells, (2) wt1 gene expression is not an independent prognostic factor for CR, DFS or OS in childhood ALL treated by an intensive therapy protocol. PMID- 10480288 TI - Induction of cytosolic phospholipase A2 and prostaglandin H2 synthase-2 by lipopolysaccharide in human polymorphonuclear leukocytes. AB - Polymorphonuclear leukocytes (PMNs) produce arachidonic acid (AA) metabolites including thromboxane A2 (TXA2). These cells are the first line of defense against bacterial invasion, which often causes endotoxin shock. TXA2 which plays an important role in the pathogenesis of endotoxin shock is synthesized by three consecutive enzyme activation, cytosolic phospholipase A2 (cPLA2), prostaglandin H2 synthase (PHS type 1 and type 2) and TXA2 synthase. Among them, cPLA2- and PHS 2 activity is known to be transcriptionally and/or posttranscriptionally up regulated by various bioactive substances including lipopolysaccharide (LPS), a bacterial endotoxin, in many cell types. We investigated the action of LPS on TXA2 synthesis in human PMNs. A23187-stimulated production of thromboxane B2 (TXB2, a stable metabolite of TXA2), assayed by specific radioimmunoassay (RIA), was significantly increased from 566.7+/-44.1 pg/10(6) cells to 966.7+/-44.1 pg/10(6) cells (p<0.05) after 6 h-exposure to LPS at the concentration of 100 ng/ml. Messenger RNA for PHS-2, PHS-1, TXA2 synthase and cPLA2, which was assessed by reverse transcription-polymerase chain reaction (RT-PCR), was expressed in PMNs without LPS stimulation. Although PHS-2 was putatively an inducible enzyme, abundance of mRNA for PHS-2 in PMNs without LPS stimulation was detectable. Messenger RNA abundance for PHS-2 and cPLA2, but not for PHS-1 and TXA2 synthase, was enhanced by LPS-treatment, indicating that the increased production of TXB2 was attributable to the up-regulation of cPLA2 and PHS-2. We conclude that (1) PHS-2 plays a more important role than PHS-1 in the production of TXA2 in human PMNs and (2) TXA2 synthesis in human PMNs is transcriptionally up-regulated by new induction of cPLA2 as well as PHS-2, when the cells encounter endotoxin producing bacteria. PMID- 10480289 TI - Low-dose oral iron absorption test in anaemic patients with and without iron deficiency determined by bone marrow iron content. AB - The low-dose oral iron absorption test (OIAT) was performed in 85 consecutive anaemic patients referred for bone marrow examination in order to investigate the ability of the test to predict bone marrow iron stores and to differentiate between different categories of anaemia. Eight patients were excluded for technical reasons. Test results from 77 patients are presented as Cmax (micromol/l): the maximum increase in S-iron measured during a 3 h period after administration of 10 mg oral iron sulfate. Iron deficiency was defined as the absence of stainable iron in bone marrow aspirates. Cmax was higher in 46 iron deficient patients [3 (median); 0 and 13 (1st and 3rd quartiles); 0-40 (range)] than in 31 non-iron-deficient patients (0; 0 and 2; 0-4) (P<0.01). 27 patients had primary bone marrow disease, 25 patients had absent bone marrow iron stores accompanied by inflammation, 17 patients had anaemia of chronic disease (ACD) and 8 patients had uncomplicated iron deficiency anaemia (IDA). Patients with IDA had higher Cmax (15; 13 and 28; 6-40) than patients with ACD (1; 0 and 2; 0-3), and than the 69 non-IDA patients (1; 0 and 3; 0-19) (P<0.001). Cmax values above 5 micromol/l always indicated absent bone marrow iron stores. PMID- 10480290 TI - Induction of differentiation in U-937 and NB4 cells is associated with inhibition of tissue factor production. AB - Tissue factor (TF) production is under strict control in mature monocytic cells. However, constitutive expression of TF can be found in myelomonocytic cells and in haematopoietic cells arrested at an early stage of differentiation. In this paper we show that TF expression is down-regulated during the monocyte/granulocyte differentiation process, using the human monoblastic U-937 and the acute promyelocytic leukaemia NB4 cell lines as models. Expression of TF mRNA, protein and procoagulant activity (PCA) was constitutively high in untreated cells. Exposure of U-937 cells to 1alpha,25-dihydroxycholecalciferol (VitD3) and all-trans retinoic acid (ATRA) resulted in down-regulation of TF expression and PCA. In NB4 cells induction by ATRA, but not VitD3, resulted in the down-regulation of TF expression and PCA. Consistent with this, induction of terminal differentiation, as confirmed by the expression of differentiation associated antigens and cell cycle arrest, was inversely correlated to TF expression in U-937 and NB4 cells. Moreover, terminally differentiated U-937 cells retained the capacity to respond to inflammatory mediators, i.e. lipopolysaccharide and interferon-gamma, by a rapid increase in TF expression. In conclusion, we show that not only ATRA but also VitD3 is a potent suppressor of monocytic TF expression and thus might have potential clinical use for the treatment of coagulopathies. PMID- 10480291 TI - High incidence of maternal HLA A, B and C antibodies associated with a mild course of haemolytic disease of the newborn. Group for the Study of Protective Maternal HLA Antibodies in the Clinical Course of HDN. AB - Mild courses of haemolytic disease of the foetus or newborn (HDN) due to Rh (D) blood group antibodies are associated with and may therefore be ameliorated by maternal antibodies reacting with human leucocyte antigens (HLA) of the child, an observation drawn from our own earlier data (Neppert J, Kissel K. Lancet 1992;339:1481). This study (i) corroborates this association; (ii) reveals shortcomings in the published data; and (iii) examines the characteristics of HDN cases when these shortcomings have been rectified. Samples from 51 women with antibodies against their child's blood group antigens of the Rh system were analysed for HLA A, B, C and DR antibodies during parturition. The mothers were divided into two groups, either severe or mild, dependent upon the clinical course of the HDN, and the incidence of HLA antibody production was determined for each group. HLA A, B, C and/or DR antibodies were detected in 85.2% of those women whose children had a mild course of HDN prenatally or perinatally (n=27). This is statistically greater than the incidence of 50.0% (Fisher's exact test: p=0.014) found in the group of women whose children had a severe HDN either pre- or perinatally (n=24) and is greater than the 35% (n=20; p=0.0001) found in women without Rh or other irregular antibodies. HLA DR antibodies were detected in three cases. The findings support our hypothesis that maternal anti HLA A, B and C antibodies may protect against a potential severe HDN. We therefore assume that those women will benefit who have already had a child with a severe HDN and in whom HLA antibodies were not previously detected, if HLA antibody production is provoked by subcutaneously inoculating with the father's leucocytes before or at the beginning of the new pregnancy. PMID- 10480292 TI - Long-term results of 60 patients with pathologic stage I & IIA Hodgkin's disease treated with exclusive mantle radiation therapy. AB - Between January 1972 and December 1982 60 patients with pathological stage IA and IIA Hodgkin's disease (HD) were submitted to Mantle irradiation only. Twenty-five were in stage I (32.1%) and 35 in stage II (67.9%). All patients were submitted to staging laparotomy. Cases with large mediastinal mass were excluded from this series. Delivered doses were 44 Gy in involved areas, 40 Gy on the mediastinum and 36 Gy on uninvolved sites. Twenty-four patients in stage I (96%) and 33 in stage II (94.2%) obtained complete remission. Actuarial 10- and 20-yr overall (OS) rates were 86% and 79.1%, respectively. Event-free (EFS) and relapse-free (RFS) survival rates at 10 and 20 yr were 67.5% and 62.1%, respectively. The occurrence of disease relapse resulted in the only statistical significant prognostic factor for OS in both univariate and multivariate analysis. Distant and extranodal recurrences were significantly (P<0.01) related to a reduced OS. On multivariate analysis stage was the only determinant factor for increased RFS. Extended field RT proved to be an effective curative modality for stage I HD patients, whereas 15 out of 33 patients in stage II relapsed requiring salvage therapy. Long-term analysis of survival and treatment-related morbidity rates will improve our knowledge and assist the physicians to choose the therapeutic option to offer to HD patients. PMID- 10480293 TI - Beneficial effect of highly active antiretroviral therapy (HAART) in reducing both HIV viral load and monoclonal gammopathy. PMID- 10480294 TI - Haemoglobin F (HbF) levels in sickle-cell anaemia patients homozygous for the Bantu haplotype. PMID- 10480295 TI - Inherited combined deficiency of proteins C and S. PMID- 10480296 TI - Recent advances in the diagnosis of X-linked agammaglobulinemia. PMID- 10480297 TI - Peripheral neuropathies in lupus patients with anti-GM1 antibodies. PMID- 10480298 TI - Role of hyperinsulinemia in atherosclerotic coronary arterial disease: studies of semi-quantitative coronary angiography. AB - OBJECTIVE: To evaluate the role of insulin resistance on coronary atherosclerosis, angiographic semiquantitative scores of coronary stenosis and calcification were evaluated. SUBJECTS AND METHODS: Ninety-five non-diabetic subjects with coronary arterial disease were selected from our angiographic data base. Hyperinsulinemia was defined as a serum insulin level of > or = 60.4 IU/l at 120 minutes after 75 g oral glucose challenge. RESULTS: Twenty-three (24%) of the patients exhibited hyperinsulinemia. There was no difference in age or gender between the two subgroups. The incidence of hypertension, smoking habits, hypercholesterolemia, and hyperuricemia were also the same among the insulin resistance subgroups. Subjects with hyperinsulinemia had higher coronary artery scores of stenosis (11.9+/-5.6 vs 8.3+/-5.0, p<0.0001) and calcification (7.5+/ 6.3 vs 4.8+/-4.9, p<0.0001). Moreover, the stenosis score had a close linear correlation with the 120 minutes serum insulin level (r=0.266, p=0.009), but not with the fasting level. CONCLUSION: These results suggest that hyperinsulinemia is a risk for coronary arterial disease, and emphasize the severity of coronary atherosclerosis in normal glucose tolerant subjects. PMID- 10480299 TI - Factors related to impairment of activities of daily living. AB - OBJECTIVE: We examined the factors related to the impairment of activities of daily living (ADL). METHODS: ADL was evaluated by using ADL-20, which consists of 20 items from 4 major categories of activities; mobility, self-care, instrumental, and communication. The patients' gender, birth date, clinical diagnosis, past history, life styles, physical findings, laboratory data, and details of therapy were also recorded. Patients A total of 1,163 outpatients aged 50 years or older were included. Data from 1,093 patients were analyzed. RESULTS: We divided the subjects into two groups; Group I having full marks of ADL-20 (n=582) and group II exhibiting an impairment of ADL (n=511). Multiple logistic analysis revealed that in both sexes age and stroke were common independent factors related to the impairment of ADL. Other factors associated with impairment of ADL were smoking in men and presence of proteinuria in women. The presence of hyperlipidemia was associated with preservation of the ADL in women. CONCLUSION: The results demonstrated significant associations of smoking in men and the presence of proteinuria in women with the impairment of ADL in elderly Japanese outpatients. There appears to be a sex difference in the risk factors of impairment of ADL. PMID- 10480300 TI - Specialty-related disparities of readmission in patients with chronic heart failure: the importance of hospital-clinic cooperation. AB - OBJECT: The purpose of this study was to elucidate differences in readmission rates and late outcome in outpatients with chronic heart failure treated in different clinical settings. PATIENTS AND METHODS: This study included 65 consecutive patients who were admitted to our CCU due to acute heart failure for the first time and discharged from our institution. After their discharge, 31 were cared for by a cardiologist in the outpatient clinic of our institution (group A) and the other 34 were cared for by a general practitioner in a clinic (group B). The various findings during the acute phase and the follow-up period were retrospectively compared between the two groups. In addition, the incidence of unexpected readmission and prolonged outcomes were compared between the two groups. RESULTS: The patients in group B were older than those in group A, but no other differences were noted in patient characteristics. More patients in group A required more than one hospitalization within 6 months from discharge (group A, 35.5%; group B, 8.9%, p<0.01; follow-up period, 17.1+/-5.9 months). There was no difference in the survival rate between the groups. CONCLUSION: We concluded that stabilized outpatients should receive comprehensive care from a general practitioner to avoid the need for readmission after discharge. PMID- 10480301 TI - Effects of combined emergency percutaneous cardiopulmonary support and reperfusion treatment in patients with refractory ventricular fibrillation complicating acute myocardial infarction. AB - OBJECT: We conducted a prospective study to determine whether or not combined emergency percutaneous cardiopulmonary support (PCPS) and coronary reperfusion treatment are useful for acute myocardial infarction (MI) patients with unsynchronized electric shock-resistive ventricular fibrillation (VF). PATIENTS AND METHODS: Thirty-two acute MI patients who lapsed into the refractory VF were entered into the study. Group 1 consisted of 19 patients with VF outside the hospital, and Group 2 consisted of 13 patients with VF immediately after arrival at the hospital. The primary endpoint was successful reperfusion, return of spontaneous circulation and good recovery without neurologic disability. RESULTS: The infarct-related arteries showed a significant difference between Groups 1 and 2. However, the two groups had similar rates of successful reperfusion (84.6% vs 94.7%, respectively) and return of spontaneous circulation (89.5% vs 84.6%, respectively). The rates of good recovery were similarly low in both groups (5.3% vs 15.4%, respectively). CONCLUSION: Combined emergency PCPS and reperfusion treatment produced high return of spontaneous circulation, however the neurologic outcome was low. PMID- 10480302 TI - Relationship between the clinical efficacy of pentoxifylline treatment and elevation of serum T helper type 2 cytokine levels in patients with human T lymphotropic virus type I-associated myelopathy. AB - OBJECT: Previously, we reported the efficacy of pentoxifylline (PTX) treatment in human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (HAM). Here, we clarify the relationship between the clinical efficacy of PTX treatment and elevation of T helper type 2 (Th2) cytokine levels in HAM patients. PATIENTS AND METHODS: PTX (300 mg) was administered daily by the oral route to 12 HAM patients for 4 weeks. We assessed the relationship between the changes in neurological status (motor disability scores, the degree of spasticity on neurological examination, and the time required to walk 10 m) and the changes in serum and cerebrospinal fluid (CSF) levels of interferon-gamma (IFN-gamma) as a Th1 cytokine and interleukin-4 and -10 (IL-4 and -10) as Th2 cytokines measured by an EASIA (enzyme-amplified sensitivity immunoassay) kit. RESULTS: PTX treatment induced incremental increases in the levels of IL-4 and IL-10 in both sera and CSF of 6 HAM patients. Clinical improvement was associated with this elevation in IL-4 and IL-10. PTX treatment also induced a decrease in IFN-gamma levels in the sera of 6 HAM patients, but this was not correlated with clinical improvement. CONCLUSION: These results suggest that the correction of the immunological imbalance in Th1 to Th2 cytokine responses, with upregulation of IL-4 and IL-10, may account for the clinical improvement in HAM patients treated with PTX. PMID- 10480303 TI - Atypical X-linked agammaglobulinemia diagnosed in three adults. AB - OBJECT: X-linked agammaglobulinemia (XLA) is one of the most common humoral immunodeficiencies characterized from childhood by the absence of peripheral B lymphocytes, reduced levels of serum immunoglobulins and recurrent and severe bacterial infections. These characteristics are the result of Bruton's tyrosine kinase (Btk) protein deficiency in peripheral B lymphocytes. In addition to typical XLA, several atypical cases have been recognized, who exhibited mild or even no clinical symptoms, although they were definitely deficient in Btk protein (atypical XLA). In these patients peripheral B lymphocytes and serum immunoglobulins (Igs) are detectable though at a lower level than in normal people. To clarify the discrepancies between the Btk gene mutations and the phenotypes more atypical patients should be examined. In this study we evaluated the cytoplasmic Btk protein in peripheral monocytes of some hypogammaglobulinemia adults by means of flowcytometric analysis. MATERIALS AND METHODS: Heparinized venous blood samples were collected from some hypogammaglobulinemia adults. Mononuclear cells were separated from their blood and first reacted with a phycoerythrin-labeled CD14 monoclonal antibody (MoAb) (staining of monocyte membrane). Next, the cells were fixed and permeabilized. And then these permeabilized cells were reacted with an anti-Btk MoAb (staining of cytoplasmic Btk protein) and incubated with a FITC-conjugated goat antimouse IgG1. The double stained cells were analyzed on a flowcytometer. RESULTS AND CONCLUSION: By means of flowcytometric analysis we diagnosed three hypogammaglobulinemia adults as XLA, who did not show typical clinical progress of XLA. Advancements in diagnostic methods has facilitated a prompt and definite diagnosis of this disease. PMID- 10480304 TI - Spontaneous hemothorax secondary to immature teratoma of the mediastinum. AB - Spontaneous hemothorax in a 20-year-old boy was caused by rupture of an immature teratoma of the mediastinum. The tumor bled spontaneously into the right pleural space. This life-threatening complication necessitated emergency surgery. The unusual cause and the interesting clinical course of spontaneous hemothorax are described. PMID- 10480305 TI - Systemic lupus erythematosus demonstrating serum anti-GM1 antibody, with sudden onset of drop foot as the initial presentation. AB - In systemic lupus erythematosus (SLE), peripheral neuropathies are relatively uncommon and rarely present as the initial symptom. We herein describe a 61-year old woman who developed a sudden onset of drop foot, which was indistinguishable from Guillain-Barre syndrome based on the clinical symptoms alone. Antibodies against ganglioside GM1 were detected in the serum, while no antibodies to Campylobacter jejuni were observed. An electrophysiological study showed axonal impairment rather than demyelination. A pathological examination of a sural nerve biopsy specimen and further laboratory examinations suggested the observed peripheral neuropathies to have arisen due to lupus vasculitis. The serological activities of SLE responded well to treatment with corticosteroids, mizoribine and immunoadsorption therapies, however, the drop foot symptoms did not change remarkably. PMID- 10480306 TI - Pulse intravenous cyclophosphamide treatment for steroid-resistant interstitial pneumonitis associated with polymyositis. AB - Interstitial pneumonitis (IP) is a serious complication in polymyositis/dermatomyositis (PM/DM), leading to significant morbidity or mortality. Here, we report the successful treatment by pulse intravenous administration of cyclophosphamide in the early course of lung involvement in PM, and with subsequent low-dosage oral administration of azathioprine in a patient with steroid-resistant IP associated with PM/DM. Although the precise pharmacological mechanism induced by cyclophosphamide in this disease remains unclear, such a cytotoxic drug raises the possibility of control of steroid resistant PM/DM-associated IP when used in the early course of IP. PMID- 10480307 TI - Polyangiitis overlap syndrome with eosinophilia associated with an elevated serum level of major basic protein. AB - Polyangiitis overlap syndrome is a new disease entity and the reported cases in the literature are still limited. We describe a female patient presenting with finger ulcers, skin eruptions, pleural effusion, interstitial pneumonia and eosinophilia. Skin biopsy showed systemic small-sized angiitis and thrombosis. She was diagnosed as having polyangiitis overlap syndrome and was successfully then treated with corticosteroid. It is also of interest that the disease activity was correlated with the number of eosinophils in peripheral blood. The measurement of the serum level of major basic protein released from eosinophils functioning as a coagulant indicated the possible association of eosinophilia with thrombosis and polyangiitis. PMID- 10480308 TI - Invasive pulmonary aspergillosis diagnosed early by polymerase chain reaction assay. AB - We compared the usefulness of a polymerase chain reaction (PCR) assay for the early diagnosis of invasive pulmonary aspergillosis with the serodiagnosis of sufficient concentrations of galactomannan using the same serum samples. A patient was treated with prednisolone for the management of hepatitis. Computed tomography (CT) scan of the chest showed the nodular shadow with a cavity containing a clear fungus ball. DNA of Aspergillus spp. from a serum sample was detected and using the same serum sample, both latex agglutination and sandwich enzyme-linked immunosorbent assay (ELISA) of galactomannan were negative. PCR assay provides an early diagnosis of invasive pulmonary aspergillosis compared with ELISA of galactomannan. PMID- 10480309 TI - Toxic shock-like syndrome resembling phlegmasia cerulea dolens. AB - A case of a 71-year-old Japanese woman with toxic shock-like syndrome is reported. She was admitted to the hospital because of swelling of the right leg. On admission, right lower leg was erythematous and swollen with an erosive lesion. On the second day, she rapidly fell into shock. Phlegmasia cerulea dolens caused by massive iliofemoral venous thrombosis was suspected because she had a history of deep venous thrombosis. But deep venous thrombus was not detected by venography at emergent surgery for thromboembolectomy. She died 38 hours after admission despite maximal supportive therapy. Group A beta-hemolytic streptococci were isolated from blood culture and soft tissue after the patient died. PMID- 10480310 TI - Gastrectomy is a risk factor for alcoholism. PMID- 10480311 TI - Red blood cell metabolism and hemoglobin oxygen affinity. Effect of vinburnine on normobaric hypoxic rats. AB - The influence of Vinburnine on red blood cell metabolism and hemoglobin oxygen affinity was investigated in normobaric rats (FiO2: 10%). Two periods of hypoxic exposure were performed (10-21 d). In each experimental series, rats were divided into two groups: Treated rats received a daily intraperitoneal injection of drug (4 mg/kg). Control rats in the same conditions of hypoxia received an isotonic saline solution. For a 10 d exposure period, Vinburnine does not affect red blood cell metabolism nor hemoglobin oxygen affinity. At 21 d period exposure, 2, 3 diphosphoglycerate (2, 3 DPG) and ATP amounts increase in treated rats. The rate of increase was 10% (p<0.05) and 28% (p<0.01), respectively. Red blood cell metabolism effect of Vinburnine was not accompanied by a modification in affinity hemoglobin oxygen (Hb-O2); no statistically significant difference was observed between treated rats and control rats concerning p50 (partial pressure oxygen at half hemoglobin saturation). Results suggest that Vinburnine has a metabolic effect corresponding to a glycolysis anaerobic stimulation, which can improve oxygen delivery to tissues and could explain the favorable hemoreological action of Vinburnine observed in a previous investigation. PMID- 10480312 TI - Long-term n-3 fatty acid deficiency induces no substantial change in the rate of protein synthesis in rat brain and liver. AB - The influence of long-term n-3 fatty acid deficiency on the rate of protein synthesis in rat brain and liver was investigated in relation to learning behavior or a presumed survival time-shortening factor (SSF) in rapeseed oil, using a large-dose [3H]phenylalanine (Phe) injection method. When Wistar rats were made n-3 fatty acid-deficient by feeding a safflower oil (alpha-linolenate deficient) diet for 2 generations, conditions under which the safflower oil group had been shown to exhibit altered learning behaviors, compared with the perilla oil group, no significant changes in the rate of protein synthesis were observed compared with the perilla oil (alpha-linolenate-sufficient) or rapeseed oil (alpha-linolenate-sufficient but SSF-containing) groups. However, the rapeseed oil group had a reduced specific radioactivity of free Phe in the cerebral cortex, compared with the safflower oil group. In contrast to the reported observation of very long-term n-3 fatty acid deficiency inducing an almost 2-fold increase in the rate of protein synthesis in the brain, our results indicate that altered learning behavior resulting from n-3 fatty acid deficiency in rats is not associated with any substantial changes in the rate of protein synthesis in the brain. PMID- 10480313 TI - A vanadyl sulfate-bovine serum albumin complex stimulates the release of lipoprotein lipase activity from isolated rat fat pads through an increase in the cellular content of cAMP and myo-inositol 1,4,5-trisphosphate. AB - A vanadyl sulfate-bovine serum albumin complex (vanadyl-BSA) prolonged the stability of the V4+ oxidation state, although vanadyl alone can readily change the oxidation state from V4+ to V5+ under physiological conditions. Vanadyl-BSA stimulated the release of lipoprotein lipase (LPL) activity from isolated rat fat pads and increased the cellular LPL activity in a time-dependent manner. These effects were independent of protein synthesis. Propranolol, quin 2-AM, ruthenium red, and neomycin all inhibited LPL release more potently than the increase in activity. In contrast, potent inhibition of the increase effect was observed with genistein and wortmannin. Short-term incubation of the fat pads with vanadyl-BSA showed a transient increase in the cellular content of cAMP and myo-inositol 1,4,5-trisphosphate (IP3), which was inhibited by propranolol and neomycin, respectively. These results suggest that vanadyl-BSA stimulates the release of LPL activity through an increase in the cellular content of cAMP and IP3, leading to an increased intracellular Ca2+ concentration, and that it also increases cellular LPL activity via process(es) sensitive to genistein and wortmannin. PMID- 10480314 TI - Influence of various bile acids on the metabolism of glycyrrhizin and glycyrrhetic acid by Ruminococcus sp. PO1-3 of human intestinal bacteria. AB - Ruminococcus sp. PO1-3, an intestinal bacterium isolated from human feces, metabolized glycyrrhizin (GL) to glycyrrhetic acid (GA) and GA to 3-oxo glycyrrhetic acid (3-oxo-GA) and possessed GL beta-D-glucuronidase and 3beta hydroxysteroid dehydrogenase (3beta-HSD) involved in the metabolism of GL. This bacterial growth was enhanced by GL at a concentration of 0.4 mm and was suppressed by GA at concentration of 1.0 mM. Chenodeoxycholic acid, deoxycholic acid and lithocholic acid among the bile acids added to this bacterium suppressed the growth and GL beta-D-glucuronidase activity and 3beta-HSD activity incident to it at a concentration of 1.0 mM, while cholic acid, hyodeoxycholic acid and glycine and taurin conjugates of cholic acid, chenodeoxycholic acid, deoxycholic acid and lithocholic acid had almost no effect on this bacterium at a concentration of 0.2 to 1.0 mm. However, these enzyme activities of this sonicated bacteria were inhibited by all of these bile acids. Although each bile acid and GL added to bacteria at the same time suppressed the growth and the amount of metabolite GA by all bile acids used except cholic acid, taurocholic acid and taurodeoxycholic acid with GL, a combination of each bile acid and GA eased the growth inhibition caused by GA at a concentration of 0.2 mM and enhanced the amount of metabolite 3-oxo-GA by the glycine conjugate of bile acids with GA. GL or GA added after 6 h culture with each of these bile acids and bacteria was metabolized to a relatively large amount of GA by chenodeoxycholic acid and lithocholic acid and their glycine and taurine conjugates, glycocholic acid and taurodeoxycholic acid, or had almost no effect on the amount of metabolite 3-oxo-GA, respectively. These results showed that although GL added after the exposure to bile acid and GA and bile acid added at the same time as bacteria had different bile acid action, these conditions enhanced the amount of metabolite GA from GL and metabolite 3-oxo-GA from GA. PMID- 10480315 TI - Abundance of low molecular weight phosphotyrosine protein phosphatase in the nerve-ending fraction in the brain. AB - The distribution of low molecular weight phosphotyrosine protein phosphatase (LMW PTP) in subcellular fractions of rat brain tissue was investigated by immunoblotting analysis using anti-LMW-PTP antibody. The enzyme was detected in the 105000 g precipitate in addition to the supernatant of brain homogenate, even after the precipitate was extensively washed, and was abundant in the particulate fraction of nerve endings. Nerve ending LMW-PTP was effectively solubilized by 1% Triton X-100 or 1% deoxycholate, though the enzyme was solubilized by thorough sonication. Two forms of LMW-PTP, designated as LMW-PTP-I and -II, were separated from the nerve ending-rich fraction by chromatofocusing. Nerve endings PTP-I and II were different in molecular weight, isoelectric point and susceptibility to activators and inhibitors. The properties of nerve endings LMW-PTP-I and -II were similar to those of cytosolic LMW-PTP-I and -II. The abundance of LMW-PTP in nerve endings as well as in the cytosol suggests that this enzyme plays an important role in synaptic function. PMID- 10480317 TI - Suppressive effect of genistein on rat bone osteoclasts: apoptosis is induced through Ca2+ signaling. AB - The effect of genistein on osteoclast-like multinucleated cells (MNCs) from rat femoral tissues was investigated. When rat marrow cells were cultured for 7 d in a medium containing either vehicle, parathyroid hormone (10(-8) M) or prostaglandine E2 (10(-6) M), the formation of tartrate-resistant acid phosphatase (TRACP)-positive MNCs was significantly increased. This increase was markedly weakened by the presence of genistein (10(-6) or 10(-5) M). The bone cells isolated from rat femoral tissues were cultured for 48 h in an alpha minimal essential medium (5% fetal bovine serum) containing either vehicle or genistein (10(-8) to 10(-5) M). Osteoclasts were estimated by staining for TRACP, a marker enzyme of osteoclasts. The presence of genistein caused a significant decrease in the number of osteoclasts. Such a decrease was also seen in the presence of calcitonin (10(-10) to 10(-8) M), dibutyryl cyclic adenosine 5' monophosphate (DcAMP; 10(-6) and 10(-5) M), calcium chloride (10(-4) and 10(-3) M) or daidzein (10(-7) to 10(-5) M). The suppressive effect of genistein (10(-5) M) was not further enhanced in the presence of calcitonin (10(-8) M), DcAMP (10( 5) M), or calcium chloride (10(-3) M), and was completely abolished by the presence of dibucaine (10(-6) M) or staurosporine (10(-7) M), both of which are inhibitors of Ca2+-dependent protein kinases. Ca2+ ionophore (A23187; 10(-6) M) induced a remarkable decrease in the number of osteoclasts in the absence or presence of genistein (10(-5) M) or calcium chloride (10(-3) M). The present study demonstrates that genistein has a direct suppressive effect on osteoclasts in vitro, suggesting that the isoflavone may induce apoptosis which is mediated through the pathway of intracellular Ca2+ signaling. PMID- 10480316 TI - Decavanadate inhibits the cell-free activation of neutrophil NADPH oxidase without affecting tyrosine phosphorylation. AB - NADPH oxidase was activated by arachidonate in a cell-free system consisting of membrane and cytosol fractions prepared from guinea pig neutrophils. Vanadate apparently inhibited the NADPH oxidase activity in the cell-free system (IC50=2 microM) without phosphotyrosine accumulation. The pH dependency and stability of the inhibitory effect observed for vanadate solution indicated that decavanadate, an isopolyanion of vanadate, was responsible for the inhibition. Pervanadate (vanadyl hydroperoxide) also inhibited the oxidase activity but at a higher concentration (IC50=0.2 mM). Decavanadate lowered the Vmax but did not affect the Km value of NADPH oxidase for NADPH. Decavanadate inhibited the activation process of NADPH oxidase but not the oxidase activity itself. Decavanadate pretreatment of membrane and cytosol fractions irreversibly decreased the abilities of both fractions to activate NADPH oxidase in the cell-free system. Translocation of p47-phox, one of the cytosolic activation factors of NADPH oxidase, from cytosol to membrane, was little affected by decavanadate. These results suggest that decavanadate inhibits the activation of NADPH oxidase in the cell-free system without affecting the phosphotyrosine phosphatase, and that decavanadate can bind to both the membrane and cytosolic activation factors when they are in a dormant state, but not to the active oxidase complex. PMID- 10480318 TI - Allopurinol increases ear swelling and mortality in a dinitrofluorobenzene induced contact hypersensitivity mouse model. AB - The immunomodulatory effects of allopurinol were investigated in a mouse contact hypersensitivity model. Allopurinol caused a time- and dose-dependent lethal effect in dinitrofluorobenzene (DNFB)-sensitized mice. Furthermore, allopurinol markedly increased ear swelling in the remaining mice. In contrast, TMX-67, a newly synthesized xanthine oxidase/xanthine dehydrogenase (XOD/XDH) inhibitor, had almost no effect on DNFB-sensitized mice. Allopurinol reduced both the spleen weight and white blood cell count in DNFB-sensitized mice without affecting the T cell subset of splenocytes. The production of interferon (IFN)-gamma, in the splenocytes of DNFB-sensitized mice was reduced by allopurinol administration. Death due to allopurinol was much lower in the non-sensitized mice than in the DNFB-sensitized mice. These findings indicate that allopurinol may interact with DNFB to enhance its toxicity and allopurinol might also modulate or enhance the inflammatory effect of DNFB. Also, DNFB may cause metabolic alterations via inflammation, leading to enhanced allopurinol toxicity. PMID- 10480319 TI - Examination of local anti-inflammatory activities of new steroids, hemisuccinyl methyl glycolates. AB - To overcome the side effects of steroids, novel steroid-17-yl methyl glycolates with succinyl group at C-20, derived from prednisolone and dexamethasone were prepared based on a concept of the antedrug. Their topical anti-inflammatory activity and systemic effects were evaluated through croton oil-induced ear edema and paper disk granuloma bioassay. Among them, (20S)-succinyl dexamethasone derivatives (7 and 11) indicated more potent anti-inflammatory activity than the parent dexamethasone and did not show corticosteroidal side effects of thymic, adrenal involution or body weight loss. Both 7 and 11 were immediately metabolized into active compound 3 and this metabolite was eliminated with mean half-lives of 0.786 to 0.866 h in rat serum. Our findings suggest that these two compounds might be candidates as the novel steroids, and that that introduction of the succinyl group into the methyl glycolates at C-20 is useful to avoid suppression of organs due to the side effects of corticosteroids. PMID- 10480320 TI - Study of the pharmacological effect of the bile salt, sodium scymnol sulfate, from Rhizoprionodon acutus. III. Protective effect of scymnol against vascular endothelial cell damage in a rat peripheral arterial occlusion model. AB - The prophylactic action of scymnol in a rat peripheral arterial occlusion model, involving injection of 5% lactic acid into the femoral artery, was investigated. Increases in serum lactate dehydrogenase (LDH), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) activities and in plasma levels of thrombin and antithrombin III complex (TAT) were observed in this model 3 h after injection of lactic acid. Changes in LDH activity were characterized by increases in isozymes LDH4 and LDH5 and an elevated LDH4/LDH5 ratio. The ratio of the LDH4 to LDH5 increments was similar to that seen in a rat endothelial cell culture. Oral preadministration of scymnol had a preventive effect on the development of lesions in this model. It significantly reduced the LDH4 and LDH5 activity, the LDH4/LDH5 ratio and the TAT levels dose-dependently over the range 1, 3 or 10 mg/kg, compared with the values in control rats. However, its administration after lactic acid injection, or to sham-operated rats was ineffective, even at a dose of 10 mg/kg. The effects of scymnol were also compared with those of ticlopidine and argatroban. The findings show that scymnol may be useful in preventing thrombotic peripheral arterial occlusive disorders and that it potently protects endothelial cells against lactic acidosis in this model. PMID- 10480321 TI - Study of the pharmacological effect of the bile salt, sodium scymnol sulfate, from Rhizoprionodon acutus. IV. Effects of naturally occurring bile alcohols, bile acids and their conjugates on lesion development and vascular endothelial cell injury in a rat peripheral arterial occlusion model. AB - A series of naturally occurring bile alcohols, bile acids and their conjugates has been investigated as part of our studies to develop unique anticoagulants with a potent prophylactic effect against vascular endothelial cell injury induced by lactic acidosis in vivo and in vitro. In an in vivo rat peripheral arterial occlusion model induced by lactic acid injection, oral administration of a single dose of 3 mg/kg scymnol significantly inhibited edematous swelling and development of lower limb lesions, including gangrene, and reduced changes in clotting system functions and serum lactate dehydrogenase activity. It had no effect on clotting system functions in sham-operated rats. The structure-activity relationship suggests that the [24R-(+)-5beta-cholestane-3alpha,7alpha,24,26 pento l] or [3alpha,7alpha-dihydroxy-5beta-cholanic acid] structure is important for a potent prophylactic effect following oral administration. Intravenous administration of a single dose of 0.3 mg/kg sodium (25S)-scymnol sulfate or scymnol prevented lesion progression as effectively as oral administration of scymnol. Sodium (25S)-scymnol sulfate and ursodeoxycholic acid showed clear protective effects against cultured vascular endothelial cell damage due to lactic acidosis which were dose-dependent. The above results suggest that bile steroids such as scymnol, sodium (25S)-scymnol sulfate, ursodeoxycholic acid, and chenodeoxycholic acid may play a role in protecting endothelial cells against injury caused by lactic acidosis. These compounds are candidates for novel anti ischemic drugs that act by specifically protecting vascular endothelial cells. PMID- 10480322 TI - Receptor-mediated gene transfer to cells of hepatic origin by galactosylated albumin-polylysine complexes. AB - To study whether we could enhance the liver targeting of DNA delivery via asialoglycoprotein receptors using a complex of poly-L-lysine (PLL)-condensed DNA and galactosylated bovine serum albumin (GalBSA) (GPD complex), DNA was first combined with PLL and then with GalBSA via charge interaction (GalBSA: PLL: DNA=3:0.5:1, w/w/w). This vector was characterized by dynamic laser light scattering, gel retardation assay, and electron microscopy to determine the particle size, electrostatic charge interaction, and 3-D structure. An electron micrography of GPD complex, where GalBSA: PLL: DNA=3:0.5:1 (w/w/w), showed a structure of spherical particles with a mean diameter of 145+/-24.2 nm, and the complex was positively charged. The complex was tested for specificity and efficiency of gene transfer in cultured human hepatoblastoma cell line Hep G2 and mouse fibroblast cells NIH/3T3 in vitro. Cellular uptake was specifically dependent on the abundance of galactose receptors on target cells. Hep G2 cells transfected with GPD complexed with the fusogenic peptide KALA (WEAKLAKALAKALAKHLAKALAKALKACEA) showed a significantly higher reporter gene activity than those transfected with GPD complex alone or free DNA-KALA complex. The efficiency of gene transfer mediated by GPD-KALA complex was not affected by the presence of serum in the transfection medium. The reporter gene activity in NIH/3T3 cells transfected with GPD complex was very low regardless of the presence of KALA and almost the same as that transfected with bovine serum albumin (BSA)-PLL-DNA complex (BPD complex). This gene transfer formulation may find potential applications for the gene therapy of liver diseases. PMID- 10480323 TI - Electrophysiological studies on the mechanism for enhanced intestinal transport of water-soluble compounds by antibiotic peptide bacitracin. AB - Bacitracin is an antibacterial cyclic dodecapeptide produced by Bacillus licheniformin. Besides antibacterial activity, it is reported to have a protease inhibitory activity and an absorption enhancing action. Here we determined the effects of bacitracin on transport of water-soluble dye fluoresceinisothiocyanate (FITC)-dextran across the rat intestinal mucosal membrane using an electrophysiological technique. Bacitracin enhanced the intestinal mucosal-to serosal transport of FITC-dextran in concentration-dependent and pH-dependent manners. In particular, the addition of bacitracin to the mucosal side led to a remarkable enhancement of FITC-dextran transport across the colonic membrane. Furthermore, its exhibition of transport enhancement required the existence of metal divalent cations, Ca2+ and Mg2+, in the mucosal compartment. Electrophysiological study using voltage-clamp technique revealed that a relatively lower concentration of bacitracin (5 mM) enhanced the transport of 6 carboxyfluorescein via a paracellular pathway in the colonic membrane and higher concentration of bacitracin (20 mM) affects both transcellular and paracellular routes, resulting in significant enhancement of 6-carboxyfluorescein across the colonic membrane. These findings might provide the useful information for enhancing the intestinal transport of poorly absorbable drugs by bacitracin which has multiple functions. PMID- 10480324 TI - Effects of secretable SOD delivered by genetically modified cells on xanthine/xanthine oxidase and paraquat-induced cytotoxicity in vitro. AB - We designed a new eukaryotic expression vector for secretable superoxide dismutase (SOD), which expresses human SOD cDNA by fusing it to 1 connecting amino acid and the signal peptide DNA sequence of the human interleukin-2 (IL-2) gene (IL-SOD(2) cDNA). The ILSOD(2) cDNA constructed by PCR-based gene expression was ligated into the multicloning site of the pRc/CMV plasmid (pRc/CMV-ILSOD(2)). Rat lung epithelial-like cells (L2 cells) and rat skin fibroblasts (FR cells) were transfected with pRc/CMV-ILSOD(2) by lipofection. The extracellular SOD activities of IS(2)-L2 cells (L2 cells transfected with pRc/CMV-ILSOD(2)) and IS(2)-FR cells (FR cells transfected with pRc/CMV-ILSOD(2)) were 2-3 times higher than those of host cells. Initially, we investigated the protective effect of extracellular SOD secreted from these transformed cells (IS(2)-L2 and IS(2)-FR cells) on extracellular superoxide anion (xanthine/xanthine oxidase; X/XO treatment)-induced cytotoxicity in normal cells. The sensitivities of these transformed cells to X/XO-induced cytotoxicity was decreased significantly as compared with that of host cells. Although, the conditioned medium from IS(2)-L2 and IS(2)-FR cells protected against X/XO-induced cytotoxicity, the conditioned medium from host cells (L2 and FR cells) showed no significant effects on X/XO induced cytotoxicity. Furthermore, the conditioned medium from transformed cells was more effective than that of host cells against lipid peroxidation by normal cells under conditions of oxidative stress. Second, we generated superoxide anions in the intracellular space by paraquat treatment. The transformed cells were more sensitive to paraquat-induced cytotoxicity than host cells. Following addition of catalase, the sensitivity of these genetically modified cells to paraquat became equivalent to that of host cells. These results indicated a protective effect of transfection with secretable SOD genes against extracellular superoxide anion-induced cytotoxicity although no such protective effect was observed against the intracellular cytotoxicity generated by paraquat treatment. PMID- 10480325 TI - Characteristic change of urinary elastin peptides and desmosine in the aortic aneurysm. AB - To estimate elastin metabolism in aneurysm, urinary levels of desmosine and elastin peptide in patients (n=23, range 54 to 85 years old) with aneurysm were measured by ELISA and compared between two control groups divided by age (<10 years old and >20 years old). The amounts of urinary desmosine and elastin peptide in the aneurysm group were significantly increased compared with those in the older control group (>20 years old). There was a correlation between urinary desmosine and elastin peptide in the young group. On the other hand, no such correlation was observed in the aneurysm group and the older control group. The distribution of the ratio (desmosine/elastin peptide) in the aneurysm group was different from that of the young control group. We conclude that assay of elastin peptide and desmosine in urine are useful in characterizing elastin degradation in a patient with aneurysm. PMID- 10480326 TI - Selective antagonism of endothelin ET(A) or ET(B) receptor in renal hemodynamics and function of deoxycorticosterone acetate-salt-induced hypertensive rats. AB - Effects of acute and chronic blockade of endothelin ET(A) or ET(B) receptors on renal hemodynamics and function were investigated using deoxycorticosterone acetate (DOCA)-salt-induced hypertensive rats. At 4 weeks after initiating DOCA salt treatment, intravenous bolus injection of ABT-627 (1 mg/kg), a selective ET(A) receptor antagonist, produced a sustained and significant hypotension, which was accompanied by potent renal vasodilation. When the selective ET(B) receptor antagonist A-192621 (3 mg/kg) was intravenously administered, there were marked decreases in renal blood flow and glomerular filtration rate, and increases in renal vascular resistance. A slight hypertensive effect was observed after the injection of A-192621. Next, we examined the effects of chronic treatment with ABT-627 (10 mg/kg/d, p.o., b.i.d.) or A-192621 (30 mg/kg/d, p.o., b.i.d.) for 2 weeks on renal function of animals at 2 weeks after initiating DOCA salt treatment. In the 2-week-treated DOCA-salt animals, the levels of creatinine clearance (Ccr), urinary excretion of protein (Uprotein V) and blood urea nitrogen (BUN) were not significantly different compared with those of sham operated control animals. These parameters did show statistically significant differences over the 3 to 4 weeks treatment period, between the DOCA-salt and the control animals (decrease in Ccr, and increase in Uprotein V and BUN in DOCA-salt rats), thereby indicating the gradual establishment of renal dysfunction in this hypertensive model. The DOCA-salt-induced changes in renal functional parameters were markedly attenuated by daily administration of ABT-627. In contrast, treatment with A-192621 augmented the above renal dysfunction. Our findings clearly indicate that selective blockade of the ET(B) receptor is detrimental to renal hemodynamics and the function of the hypertensive condition, and support the view that a selective ET(A) receptor antagonist is useful for treatment of subjects with mineralocorticoid-dependent hypertension. PMID- 10480327 TI - Imidapril, an angiotensin-converting enzyme inhibitor, inhibits thrombosis via reduction in aortic plasminogen activator inhibitor type-1 levels in spontaneously hypertensive rats. AB - It is known that angiotensin II (Ang II) exerts an antifibrinolytic effect by stimulating synthesis of plasminogen activator inhibitor type-1 (PAI-1), a specific inhibitor of tissue plasminogen activator (t-PA). The aim of this study was to compare the antithrombotic potency of imidapril, an angiotensin-converting enzyme (ACE) inhibitor, and candesartan, an angiotensin II type 1 (AT1) receptor antagonist, in a model of arterial thrombosis in spontaneously hypertensive rats (SHRs). Oral treatment with 5 mg/kg imidapril 1 h before induction of thrombosis resulted in a significant reduction in thrombus weight, whereas candesartan did not affect thrombus weight under the same treatment conditions. Candesartan lowered blood pressure to the same degree as in the imidapril-treated rats. Imidapril not only reduce the serum and aortic ACE activities, but also reduced aortic PAI-1 protein levels, while candesartan had no effect on theses. These results suggest that imidapril, but not the AT1 receptor antagonist, candesartan, enhances fibrinolysis via a reduction of aortic PAI-1 levels by inhibiting ACE and prevents thrombus formation in SHRs. PMID- 10480328 TI - Properties of partially purified beta-N-acetylglucosaminidase from bovine crystalline lens. AB - This paper reports that beta-N-acetylglucosaminidase from bovine lens has potent enzyme activity compared with other glycosidases in the lens. The partially purified enzyme was used to determine its physiological properties. The optimal pH and optimal temperature of this enzyme was approximately 6.3 and 40 degrees C, respectively. The apparent native molecular weight of this enzyme obtained by gel filtration chromatography was 540 kDa. Furthermore, the enzyme fraction contained 3 polypeptides with molecular weights of 28.8, 28.0 and 26.0 kDa, although it is not certain if they were one of the components of this enzyme in the current study. The Km value of this enzyme was 92.3 microM and it was inhibited strongly by HgCl2 and sodium dodecyl sulfate (SDS). PMID- 10480329 TI - Effects of phyllodulcin, hydrangenol, and their 8-O-glucosides, and thunberginols A and F from Hydrangea macrophylla SERINGE var. thunbergii MAKINO on passive cutaneous anaphylaxis reaction in rats. AB - We examined the antiallergic effects of phyllodulcin, hydrangenol, and their 8-O glucosides, and thunberginols A and F isolated from the processed leaves (Hydrangeae Dulcis Folium) and dried leaves of Hydrangea macrophylla SERINGE var. thunbergii MAKINO using the passive cutaneous anaphylaxis (PCA) reaction. With the exception of phyllodulcin, these constituents were found to significantly inhibit the PCA reaction. Although thunberginol A showed the most potent inhibitory effect, hydrangenol was considered to be the principal antiallergic component in the processed leaves, after taking into account their contents. PMID- 10480330 TI - A novel cerebroside from lycii fructus preserves the hepatic glutathione redox system in primary cultures of rat hepatocytes. AB - We previously reported the isolation of a novel cerebroside (1-O-(beta-D glucopyranosyl)-(2S,3R,4E,8Z)-2-N-palmityloc tadecasphinga-4,8-diene; LCC) from the fruits of Lycium chinense MILL. (Solanaceae) which protected primary cultured rat hepatocytes from the toxicity induced by carbon tetrachloride (CCl4). The present study was conducted to determine the mechanism(s) by which LCC might exert its hepatoprotective activity. To determine the effect of LCC on the glutathione (GSH) redox system, we measured the activities of enzymes involved in the system as well as the levels of hepatic mitochondrial GSH and malondialdehyde (MDA). The hepatotoxicant, CCl4, routinely decreased levels of total and reduced GSH. The levels of these compounds were significantly maintained at the levels of the control cultures following treatment with LCC. The decreased activities of glutathione reductase and glutathione peroxidase in CCl4-injured rat hepatocytes were significantly increased by the treatment of LCC. Furthermore, the elevated levels of MDA seen in CCl4-injured rat hepatocytes were reduced after treatment with LCC in a concentration dependent manner over a range of 1-10 microM. From these results, we postulate that LCC may preserve the hepatic mitochondrial level of GSH by scavenging reactive oxygen species produced during CCl4-induced toxicity and thereby reduce lipid peroxidation and cellular damage. PMID- 10480331 TI - Transport and recognition of aminopeptidase-resistant cellobiose-coupled tyrosylglycylglycine by intestinal Na+/glucose cotransporter (SGLT1): recognition of sugar conjugates by SGLT1 is much less restricted than transport. AB - Transport and recognition of aminopeptidase-resistant cellobiose-coupled tyrosylglycylglycine (CcpTGG) by intestinal Na+/glucose cotransporter (SGLT1) was examined in rat small intestine. Inhibitory effect of phloridzin (SGLT1 inhibitor) on the CcpTGG transport was extremely low. Concentration dependence of the CcpTGG transport was observed, but the primary component of transport was passive diffusion. However, CcpTGG significantly inhibited SGLT1-mediated transport, indicating its recognition by SGLT1. Other glucose-conjugates also inhibited SGLT1-mediated transport. These results indicate that recognition of sugar conjugates by SGLT1 is much less restricted than transport, and that it should be relatively easier to design SGLT1 inhibitors than SGLT1-transportable sugar conjugates. PMID- 10480332 TI - Conversion of dieldrin to aldrin by intestinal bacteria in rats. AB - The present study provides the evidence that dieldrin is reductively metabolized to aldrin by intestinal bacteria in rats. When dieldrin was incubated with the cecal contents of rats, aldrin, a reduced metabolite of the epoxide, was isolated from the incubation mixture. The metabolite was identified unequivocally by UV and mass spectral comparison with an authentic sample, and on the basis of its TLC and HPLC behavior. The cecal contents of rats exhibited epoxide reductase activity toward dieldrin under anaerobic conditions. However, only marginal activity was observed under aerobic conditions. Four pure strains of intestinal bacteria exhibited epoxide reductase activities to varying degrees under anaerobic conditions. The highest activity was observed in Clostridium sporogenes. Cell-free extracts of the intestinal bacteria in rat cecal contents showed reductase activity when supplemented with both NAD(P)H and FMN under anaerobic conditions. PMID- 10480333 TI - DT-diaphorase-like quinone reductase in rat plasma. AB - The present study provides the evidence that DT-diaphorase-like quinone reductase exists in rat plasma. The quinone reductase activity toward menadione was found in rat plasma in the presence of NADH or NADPH. The enzyme activity was induced by pretreatment with 3-methylcholanthrene, but was not affected by phenobarbital. The 3-methylcholanthrene-induced quinone reductase activity was separated into three fractions (F1, F2, and F3) by gel filtration, which showed NAD(P)H-linked, NADH-linked, and NAD(P)H-linked activities, respectively. F1, which was induced by 3-methylcholanthrene, was inhibited by dicumarol, and cross-reacted with rat liver DT-diaphorase antibody. PMID- 10480334 TI - Geranylgeranylacetone suppresses spontaneous apoptotic DNA fragmentation in cultured guinea pig gastric pit cells. AB - We recently found that guinea pig gastric pit cells in culture undergo spontaneous and rapid apoptotic DNA fragmentation, which may represent the rapid death of gastric pit cells in vivo. In this study, we observed that pretreatment of cells with geranylgeranylacetone, an antiulcer drug with heat-shock protein inducing activity, suppressed the spontaneous apoptotic DNA fragmentation in a dose-dependent manner. Pretreatment of cells with low concentrations of ethanol or heat-shock also prevented the spontaneous apoptotic DNA fragmentation. These observations indicate that the suppression of the apoptotic DNA fragmentation by geranylgeranylacetone involve the induction of heat-shock proteins. PMID- 10480335 TI - Losses of heterozygosity on chromosomes 17p and 9p/18q may play important roles in early and advanced phases of gallbladder carcinogenesis. AB - PURPOSE: This present study aimed to investigate the genetic changes in gallbladder carcinogenesis. METHODS: Eleven intramucosal gallbladder carcinomas were compared with 31 invasive lesions for loss of heterozygosity (LOH) on chromosomes 5q, 9p, 17p and 18q, frame-shift mutations in a ten-adenine repeat site within the gene encoding the transforming growth factor beta type II receptor (TGFbetaRII) and an eight-guanine repeat site within BAX, and point mutations in codon 12 of Ki-ras. RESULTS: The incidences of LOH in intramucosal and invasive carcinomas were 14% and 17% on 5q, 9% and 52% on 9p, 64% and 65% on 17p, and 13% and 32% on 18q. No frame-shift mutations were found at TGFbetaRII or BAX, and point mutations in codon 12 of Ki-ras were present in only 8% of the samples. Thus, LOH on 17p was by far the most frequent lesion with similar results in both intramucosal and invasive carcinomas. In contrast, the frequency of LOH on 9p and 18q was distinctly higher in invasive lesions. CONCLUSION: The present data suggest that LOH on 17p may play an important role in the evolution of gallbladder carcinoma from a relatively early phase, while LOH on 9p and 18q may play roles in progression. PMID- 10480337 TI - Telomerase is active in normal gastrointestinal mucosa and not up-regulated in precancerous lesions. AB - Telomerase is a ribonucleoprotein complex which, by de novo synthesized telomeric TTAGGG repeats, prevents telomere erosion. While telomerase is active in most cancers, conflicting results exist for normal tissues and premalignant lesions. To establish the telomerase status of normal gastrointestinal mucosa and to elucidate whether telomerase up-regulation is an early or late event in carcinogenesis, we determined the telomerase activity of 88 biopsies of normal mucosa from esophagus, stomach, and intestine and compared it with that of 21 samples of premalignant lesions and 6 adenocarcinomas using the telomere-repeat amplification protocol assay. Telomerase was found in all normal tissues, revealing most activity in esophagus (11 samples), followed by intestine (45 samples), and stomach (32 samples). In 53% of the stomach samples, enzyme activity could only be demonstrated when telomerase inhibitors were eliminated by a modified telomerase assay. In the 21 precancerous lesions (5 Barrett's esophagus, 3 stomach intestinal metaplasias, and 13 colorectal adenomas of type I/II dysplasia) a similar or even reduced telomerase activity was seen, while the adenocarcinomas showed high activity. These data demonstrate that telomerase activity is expressed in all epithelia along the gastrointestinal tract, thus confirming our previous hypothesis that telomerase is constitutively expressed in permanently renewing epithelia. Furthermore, activity was not increased in preneoplastic lesions, suggesting that telomerase up-regulation is a late event during carcinogenesis of the esophagus, stomach, and intestine. PMID- 10480336 TI - Growth inhibition of experimental pancreatic cancers and sustained reduction in epidermal growth factor receptors during therapy with hormonal peptide analogs. AB - Reduction in receptors for epidermal growth factor (EGF) in cancers appears to be one of the principal mechanisms through which peptide hormone analogs can inhibit tumor growth. In this study, hamsters with nitrosamine-induced pancreatic cancers were treated for 8 weeks with bombesin/gastrin-releasing peptide (GRP) antagonist RC-3095, somatostatin analog RC-160 or the luteinizing hormone-releasing hormone antagonist Cetrorelix, using sustained delivery systems releasing 20, 35 and 20 microg analog/ day respectively. To establish the pattern of changes in the number and affinity of EGF receptors on tumors, groups of animals were sacrificed at regular intervals during therapy. Chronic treatment with RC-3095 or Cetrorelix resulted in an early (day 10) and sustained reduction (71% or 69% respectively) in EGF receptors on pancreatic tumors. In contrast, RC-160 decreased receptor concentration by 60% only after 20 days. Among the histological characteristics of proliferation, the decrease in argyrophilic nucleolar organizer regions, but not apoptotic and mitotic indices, showed a correlation with the fall in EGF receptors. The concentration of the receptors returned to the control level 4 days after cessation of chronic treatment with RC-3095. The effect of single injections of RC-3095, RC-160 and Cetrorelix on EGF receptors was also investigated. RC-160 decreased the number of EGF receptors on pancreatic cancers by 31% 3 h after administration, but the receptors had returned to normal level at 6 h. RC-3095 and Cetrorelix caused a 67% and 59% decline, respectively, in EGF receptors only 6 h after injection and the concentration of receptors remained low for 24 h. Thus, the pattern of downregulation of EGF receptors in pancreatic cancers appears to depend on the peptide used for therapy. Since the antitumor effect may be the result of the fall in EGF receptors in cancers, information on the time course of changes in these receptors during treatment with these analogs may lead to an improvement in therapeutic regimens. PMID- 10480338 TI - Galectins-1 and -3 and their ligands in tumor biology. Non-uniform properties in cell-surface presentation and modulation of adhesion to matrix glycoproteins for various tumor cell lines, in biodistribution of free and liposome-bound galectins and in their expression by breast and colorectal carcinomas with/without metastatic propensity. AB - Protein (lectin)-carbohydrate (cellular glycoconjugate) recognition is operative in biochemical information transfer. Galectins constitute a family of endogenous galactoside-binding lectins with conserved features in the binding site. The members of this lectin category are assumed to be involved in cell adhesion and growth regulation. To assess to what extent the different modes of binding-site presentation and/or carbohydrate fine-specificities will affect aspects of galectin behavior, homodimeric cross-linking galectin-1 and monomeric chimeric galectin-3, with its collagenase-sensitive stalk linked to the carbohydrate recognition domain, were investigated. Cell-surface expression of the two galectins and accessible galectin-binding sites on various tumor cell lines was ascertained by FACScan analysis. In particular, ligand accessibility for the two galectins differed for the tested cell line types. Binding of tumor cells to laminin and plasma or placental fibronectin was generally reduced by treatment of cells or matrix with galectins. Galectin-3 was more efficient than galectin 1 at impairing laminin's potency as matrix. Cell binding of galectin-1, on the other hand, proved on average more effective for blocking cell association to fibronectins after its preincubation with cell suspensions. Differences were also apparent in the biodistribution of the galectins, where an avian homolog of galectin- served as the control to distinguish effects of spatial and sugar binding features. Histopathological analysis of lymph-node-negative and -positive breast and colorectal carcinomas (n = 180 including 60 metastatic lesions) indicated a correlation of either increased galectin-1 binding and reduced galectin-3 expression or reduced binding of both galectins with the occurrence of lymph node lesions. Together with data on the heparin-binding lectin, revealing reduced expression to be associated with a positive lymph-node status in the breast cancer group, these results can be interpreted to reflect cell-type dependent requirements of galectin ligand presentation during the metastatic cascade. By introducing mammalian lectins to lectin-histochemical studies, the detection of quantitative differences in glycosylation brings an understanding of its cell biological significance one step closer. PMID- 10480339 TI - Expression of leukemia-inhibitory factor as an autocrinal growth factor in human medulloblastomas. AB - Medulloblastoma is the most frequent pediatric brain tumor, with the capacities of rapid proliferation and intracranial dissemination. However, the factor(s) regulating medulloblastoma growth has not yet been well characterized. Leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) play different roles in the formation/progression of various embryonic and pediatric tumors, but their biological effects on medulloblastoma cells are less well known. Therefore, in vivo and in vitro expression of LIF, IL-6 and their signal transducer genes encoding LIF receptor (LIFR), IL-6 receptor (IL-6R) and gp130 in human medulloblastoma cells were investigated by multiple cellular and molecular biology approaches. The results revealed that LIF expression could be found in 26 out of 28 tumors/cell line and over 90% of the samples expressed LIFR, IL-6R and gp130. In contrast, none of the samples showed IL-6 expression. An established medulloblastoma cell line, Med-3, was used to evaluate the potential effects of LIF and IL-6 on the proliferation of medulloblastoma cells. The growth of Med-3 cells was efficiently inhibited either by anti-LIF antibody or by antisense LIF oligonucleotide. Addition of exogenous human recombinant IL-6 could dramatically enhance Med-3 cell outgrowth. Our data thus for the first time demonstrated the important role of LIF as an autocrinal and IL-6 as a paracrinal growth factor in the proliferation of medulloblastoma cells. PMID- 10480340 TI - Chemosensitivity of glioma cells in vitro: a meta analysis. AB - PURPOSE: The disappointing results of chemotherapy in glioblastoma might be caused by the choices of agents, which mostly include nitrosurea. We compared the in vitro efficacy of chemotherapeutic agents, developing a method to summarize published data. METHOD: Between 1966 and 1995 chemotherapy in glioma cells was reported in 1643 articles. Efficacy was mostly described by the drug concentration that killed 50% of the cells (LC50). It was measured with various cell-culture techniques, of which a colorimetric test [3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyltrazolium bromide] was mostly used. We calculated factors from these data to transform results to LC50 values as if the colorimetric test was used in all of them. This allowed data from all publications to be summarized in a new type of meta analysis. RESULTS: The most important agents and the average LC50 values (mg/l) were actinomycin-D 0.042, vincristine 0.075, mitoxantrone 0.12, vinblastine 0.21, doxorubicin 0.29, diaziquone 0.76, cisplatin 1.1, methotrexate 1.1, cytasine arabinoside 1.59, 5-flurouracil 2.33, bleomycin 18.6, carboplatin 29.8, carmustine 37.0, nimustine 48.9, and lomustine 76.7. The most resistant cell was SNB56, followed with increasing sensitivity by SF128 and A172, and primary cultures P497, SF210, U87MG, SF126, 9L, P540, U251MG, HU62, C6. The complete list of original data is available upon request. CONCLUSION: The efficacy of nitrosourea in vitro is low. PMID- 10480341 TI - Identification of renal-cell-carcinoma-related cDNA clones by suppression subtractive hybridization. AB - The identification of human genes expressed exclusively or preferentially in tumour tissue will be of interest for exploiting the process of oncogenesis, for diagnostic purposes and possibly for developing new strategies in cancer immunotherapy. In an attempt to identify genes differentially expressed in renal cell carcinoma we used the method of suppression subtractive hybridization, comparing renal cell carcinoma tissue with non-transformed kidney tissue. From randomly selected clones, nine were identified to be differentially expressed in renal cell carcinoma. Sequence analysis of seven out of the nine revealed substantial homologies with known genes, the remaining two cDNA clones did not match with any sequences in the GenBank/EMBL database, indicating that they may be novel genes. Notably, expression of five out of the seven known genes is associated with the malignant phenotype. Thus, suppression subtractive hybridization is an effective technique of high sensitivity for the detection of differentially expressed transcripts, not only in cell lines but also for tumour tissue. PMID- 10480342 TI - Differential expression of O6-methylguanine-DNA methyltransferase during diethylnitrosamine-induced carcinogenesis and liver regeneration in Sprague Dawley male rats. AB - Differential expression of DNA-O6MeG: protein-L-cysteine S-methyltransferase (MGMT) activity and posttranslational modification of the protein during liver regeneration and carcinogenesis were compared in Sprague-Dawley male rats after partial hepatectomy and/or single i.p injection of diethylnitrosamine (DEN, 200 mg/kg). Regenerating hepatocytes after partial hepatectomy induced MGMT transiently within 3 days; however, the induction of MGMT was persistent for 2 weeks after DEN injection, and the combined treatment of DEN and partial hepatectomy maintained the elevated MGMT level for up to 4 weeks. The increased activity was transcriptionally regulated, when analyzed by Northern blot hybridization. The major active form of MGMT protein in the partially hepatectomized or DEN-treated rats was a 26-kDa or 24-kDa species respectively, which was confirmed by Western blot analysis and gel slice assay. The biological significance of the differential induction of MGMT during partial hepatectomy or DEN-induced carcinogenesis is not obvious; however, further studies on possible posttranslational modifications of MGMT protein might shed some light on the functional aspect of MGMT induction. PMID- 10480344 TI - Cyclin D1 overexpression in invasive breast cancers: correlation with cyclin dependent kinase 4 and oestrogen receptor overexpression, and lack of correlation with mitotic activity. AB - BACKGROUND: Cyclin D1 (CCND1) and its catalytic partner cyclin-dependent kinase 4 (cdk4) are known to play important roles in the G1/S check point of the cell cycle, and CCND1 overexpression has been reported to correlate with progression and prognosis of breast cancers. Oestrogen receptor (ER) levels determine the proliferative response to oestrogen by regulating binding. It has been postulated that CCNDI and cdk4 exert effects on mammary carcinogenesis in co-operation with ER. PATIENTS AND METHODS: CCND1 and cdk4 overexpression in 117 breast cancer cases with long-term follow-up were investigated by means of immunohistochemistry and differential polymerase chain reaction (PCR), using formalin-fixed and paraffin-embedded samples, and compared with ER status and mitotic indices. Additional Western blotting and reverse transcription (RT)/PCR/Southern blot hybridization were performed for 4 breast cancer cell lines and 15 fresh-frozen breast cancer samples to confirm CCND1 and cdk4 data. RESULTS: Immunohistochemically 27 cases were CCND1-positive (23.0%), and CCND1 amplification was evident in 21 (21/86; 24.4%). The two methods in combination demonstrated 37 cases (31.6%) to be positive for CCND1 overexpression. Western blotting revealed 60% of samples of fresh tissue to overexpress CCND1, corresponding well with the results of RT-PCR. There was thus a strong discrepancy between results for paraffin block and fresh samples, probably because of the short life of CCND1. In the case of cdk4, the respective percentages for positive cases were 54.7% and 73%. CCND1 and cdk4 overexpression (P < 0.0001), and CCND1 and ER positivity (P = 0.0128) correlated. In addition, samples overexpressing CCND1, cdk4 and ER tended to have slightly lower proliferative activity than samples where these were absent. However, no association with clinicopathological parameters was evident. cdk4 overexpression had no linkage with ER status or clinicopathological status. Neither CCND1 nor cdk4 expression affected prognosis. CONCLUSION: CCND1 overexpression does not correlate with cancer progression or prognosis or with mitotic activity. The results may suggest that an excess accumulation of CCND1 in breast cancer cells tends to suppress entry into the S phase of the cell cycle. PMID- 10480343 TI - Phase I/II trial of human recombinant granulocyte-colony-stimulating factor (filgrastim) and escalating doses of cyclophosphamide, mitoxantrone, and 5-FU in the treatment of advanced breast cancer. AB - PURPOSE: We performed a phase I/II dose-escalation trial of cyclophosphamide, mitoxantrone, and 5-fluorouracil (CNF) in combination with human recombinant granulocyte-colony-stimulating factor (G-CSF, filgrastim) in patients with advanced breast cancer. The objectives of this trial were (1) to gain experience with filgrastim given to patients with advanced breast cancer and receiving standard-dose CNF, and (2) to determine the maximum tolerated dose of CNF that could be given with filgrastim support by incremental dose escalation of two components of the CNF regimen, cyclophosphamide and mitoxantrone. METHODS: Four patients who had received prior therapy for advanced disease received standard dose CNF with filgrastim support. Sequentially enrolled patients who had received no prior chemotherapy for advanced disease were treated with standard-dose CNF without filgrastim (5 patients), standard-dose CNF with filgrastim (15 patients), or were entered into sequential cohorts of 3-6 patients to be treated with increasing doses of CNF with filgrastim support (29 patients). RESULTS: The maximum tolerated doses that could be given with filgrastim support were 1500 mg/ml cyclophosphamide, 20 mg/m2 mitoxantrone, and 500 mg/m2 5-FU. Overall, 7 complete (14%) and 13 partial responses (26%) were observed. Despite the use of filgrastim, repeated cycles of CNF at doses of 2000 mg/m2 cyclophosphamide, 25 mg/m2 mitoxantrone, and 500 mg/m2 5-FU could not be given because of neutropenia and thrombopenia. Among 18 patients with bidimensionally measurable disease there were 3 complete (17%) and 5 partial (28%) responses. The median progression-free survival of all patients was 236 days (34 weeks). CONCLUSION: The use of filgrastim allows CNF to be given at approximately twice the dose intensity of "standard"-dose CNF. Because nonhematopoietic toxicity was not dose-limiting, further dose escalation of this regimen might be possible with more effective hematopoietic support. The response rate and survival of patients treated in this study were within the range expected with standard-dose chemotherapy. PMID- 10480345 TI - Mechanisms of resistance to methotrexate in childhood acute lymphoblastic leukemia: circumvention of thymidylate synthase inhibition. AB - PURPOSE: In about 25% of patients suffering from acute lymphoblastic leukemia (ALL) treatment failures occur that are most likely due to development of resistance to methotrexate (MTX). Blasts from patients with ALL were evaluated for MTX uptake, formation of long-chain MTX polyglutamates (MTX-Glu5+6), cytotoxicity and thymidylate synthase inhibition by MTX and compared to blasts from patients with acute myelogenous leukemia (AML). METHODS: Radioactively labeled MTX-Glu(n) were analyzed by means of HPLC. Thymidylate synthase activity was measured by a tritium-release assay. Cytotoxicity was determined by trypan blue exclusion. RESULTS: In most ALL blasts (n = 9) large amounts of MTX-Glu5+6 (1.06-7.03 pmol/10(7) cells) and high cytotoxicity (43.5% 92.7%) were found, while in others small amounts of MTX-Glu5+6 (0.0-0.39 pmol/10(7) cells) caused only weak cytotoxicity (6.0% 27.9%) (n = 5, 2 relapsed patients). Resistance to MTX in blasts from AML patients (n = 5) was also caused by reduced synthesis of MTX-Glu5s+6 (0.0-0.42 pmol/10(7) cells). In contrast, some ALL blasts (n = 7, 4 relapsed patients) were able to survive MTX treatment despite large amounts of MTX-Glu5+6 (1.5-5.05 pmol/10(7) cells) and extensive thymidylate synthase inhibition. CONCLUSIONS: Since the majority of ALL patients were examined at first diagnosis, an inherent mechanism of resistance seems most likely. We propose a mechanism based on the switch of thymidylate synthesis to the salvage pathway. PMID- 10480346 TI - Therapy of metastatic breast cancer with humanized antibodies against the HER2 receptor protein. AB - The HER2 protein, a member of the epidermal growth factor family, is encoded by the protooncogene c-erbB-2. Its overexpression, occurring in approximately one third of all breast carcinomas, is associated with a poor prognosis. A humanized mouse antibody against HER2 has been developed by genetic engineering. Here an unspecific human IgG was connected to the recognizing mouse IgG fragment. The allergization typical for allogeneic antibodies does not take place in this context. The effectiveness of this antibody has been confirmed by two international prospective phase III trials that tested it alone and combined with chemotherapy. Both modes of application increased the response rates and the time to progression. Side-effects were rare except for a high rate of cardiac dysfunction when the antibody was combined with anthracyclines. The effectiveness and negligible side-effects of the chimeric antibody against HER2 (Herceptin) render it a valuable tool in the treatment of breast cancer. PMID- 10480347 TI - Hypoxia induces pivotal tumor angiogenesis control factors including p53, vascular endothelial growth factor and the NFkappaB-dependent inducible nitric oxide synthase and cyclooxygenase-2. PMID- 10480348 TI - Duchenne/Becker muscular dystrophy: correlation of phenotype by electroretinography with sites of dystrophin mutations. AB - The dark-adapted electroretinogram (ERG) of patients with Duchenne and Becker muscular dystrophy (DMD/BMD) shows a marked reduction in b-wave amplitude. Genotype-phenotype studies of mouse models for DMD show position-specific effects of the mutations upon the phenotype: mice with 5' defects of dystrophin have normal ERGs, those with defects in the central region have a normal b-wave amplitude associated with prolonged implicit times for both the b-wave and oscillatory potentials, and mice with 3' defects have a phenotype similar to that seen in DMD/BMD patients. The mouse studies suggest a key role for the carboxyl terminal dystrophin isoform, Dp260, in retinal electrophysiology. We have undertaken a systematic evaluation of DMD/BMD patients through clinical examination and review of the literature in order to determine whether the position-specific effects of mutations noted in the mouse are present in man. We have found that, in man, a wider variation of DMD defects correlate with reductions in the b-wave amplitude. Individuals with normal ERGs have mutations predominantly located 5' of the transcript initiation site of Dp260. Our results suggest that the most important determinant in the ERG b-wave phenotype is the mutation position, rather than muscle disease severity. Forty-six per cent of patients with mutations 5' of the Dp260 transcript start site have abnormal ERGs, as opposed to 94% with more distal mutations. The human genotype-phenotype correlations are consistent with a role for Dp260 in normal retinal electrophysiology and may also reflect the expression of other C-terminal dystrophin isoforms and their contributions to retinal signal transmission. PMID- 10480349 TI - NPC1 gene mutations in Japanese patients with Niemann-Pick disease type C. AB - Complementary and genomic DNAs isolated from the fibroblasts of 10 Japanese (7 late infantile, 2 juvenile, and 1 adult form of the disease) and one Caucasian patient with Niemann-Pick disease type C were analyzed for mutations in the NPC1 gene. Fourteen novel mutations were found including small deletions and point mutations. A one-base deletion and a point mutation caused splicing errors. The mutations were not clustered in any particular region of the gene and were found both in and out of the transmembrane domains. Three patients were homozygous, five were compound heterozygous, and the remaining three were suspected of being compound hetrozygous with an unknown error in one of their NPC1 alleles. Of the 14 mutations, the G1553A substitution that caused a splicing error of exon 9 appeared to be relatively common in Japanese patients, because two patients were homozygous and one patient was compound heterozygous for this mutation. PMID- 10480350 TI - Human and mouse RAD17 genes: identification, localization, genomic structure and histological expression pattern in normal testis and seminoma. AB - Recently, the human orthologue to the cell cycle checkpoint genes rad17 (Schizosaccharomyces pombe) and RAD24 (Saccharomyces cerevisiae), called HRAD17, has been isolated and localized to chromosome 4. Independently, we have isolated the HRAD17 transcript and mapped it to chromosome 5q13 between the CCNB1 and BTF2p44cen genes. Furthermore, we have identified the complete exon-intron structure of HRAD17. The gene is organized into 14 exons, the translation initiation site lies within exon 2, and the stop codon within exon 14. Two further HRAD17 pseudogenes, HRAD17P1 and HRAD17P2, were identified on chromosomes 7p21 and 13q14.3, respectively, encompassing exons 3-14 and bearing 84% and 93% homology, respectively. Additionally, we have isolated the coding region of the mouse orthologue, Mrad17, and mapped it on chromosome 13 between Ccnb1 and Btf2p44, the same two genes between which it maps in human. The predicted Mrad17 polypeptide encompasses 687 amino acids and shows 89% similarity to HRAD17. Both genes are most highly expressed in testis compared to all other tissues, as shown by Northern blot hybridization. Histological studies, based on in situ hybridization with radioactively labeled antisense HRAD17 riboprobes, showed a strong expression within the germinal epithelium of the seminiferous tubuli in normal testis whereas in testicular tumors (seminomas) only weak, diffuse signals were seen. In light of the known function of the yeast orthologue at meiotic and mitotic checkpoints, as well as the strong expression in testis and weak expression in seminomas, we suggest a putative involvement of HRAD 17 in testicular tumorigenesis. PMID- 10480351 TI - BRCA1 mutations in African Americans. AB - The breast cancer predisposing gene, BRCA1, was analyzed for germline mutations in 45 African American families at high-risk for hereditary breast cancer. Patients were considered high-risk if they had a family history of the disease, early onset breast cancer, bilateral breast cancer, or breast and ovarian cancer. The entire BRCA1 coding and flanking intron regions have been examined by single stranded conformation polymorphism analysis followed by sequencing of variant bands. Eleven different BRCA1 germline mutations/variations were identified in 7 patients from the 45 high-risk families. Two pathogenic, protein-truncating mutations were detected in exon 11. A ten base pair tandem duplication, 943ins10, was present in a woman with breast and ovarian cancer whose first-degree relatives had prostate cancer. A four base pair deletion, 3450del4, was detected in a breast cancer patient with five cases of breast cancer in the family; two of the proband's sisters with breast cancer also carried the same mutation. Four amino acid substitutions (Lys1183Arg, Leu1564Pro, Gln1785His, and Glu1794Asp) and four nucleotide substitutions in intron 22 (IVS22+78 C/A, IVS22+67 T/C, IVS22+8 T/A and IVS22+7 T/C) were observed in patients and not in control subjects. One early onset breast cancer patient carried five distinct BRCA1 variations, two amino acid substitutions and three substitutions in intron 22. An amino acid substitution in exon 11, Ser1140Gly, was identified in 3 different unrelated patients and in 6 of 92 control samples. The latter probably represents a benign polymorphism. PMID- 10480352 TI - A familial case of Alzheimer's disease without tau pathology may be linked with chromosome 3 markers. AB - Alzheimer's disease is the most common form of dementia that occurs in later years. The diagnosis is confirmed by the pathological findings of betaA4-amyloid containing neuritic plaques and neurofibrillary tangles, the former being present in sufficient quantity commensurate with age. Other forms of dementia are more difficult to diagnose clinically; their pathology is noted for the lack of plaques and tangles. A patient with a family history of dementia presented with the clinical signs of Alzheimer's disease which lasted for 13 years. At autopsy the brain tissue had betaA4-amyloid-containing neuritic plaques, but no neurofibrillary tangles (i.e., the tissue was negative for staining with the tau antibody). Genetic analysis of DNA from family members revealed no linkage with chromosome 17 markers, indicating that this was not frontotemporal dementia. However, there was linkage with chromosome 3 markers. Thus, this form of Alzheimer's disease with a pathology of plaques only is linked with markers on chromosome 3. PMID- 10480353 TI - A common PEX1 frameshift mutation in patients with disorders of peroxisome biogenesis correlates with the severe Zellweger syndrome phenotype. AB - Peroxisome biogenesis disorders are a heterogeneous group of human neurodegenerative diseases caused by peroxisomal metabolic dysfunction. At the molecular level, these disorders arise from mutations in PEX genes that encode proteins required for the import of proteins into the peroxisomal lumen. The Zellweger syndrome spectrum of diseases is a major sub-set of these disorders and represents a clinical continuum from Zellweger syndrome (the most severe) through neonatal adrenoleukodystrophy to infantile Refsum disease. The PEX1 gene, which encodes a cytoplasmic AAA ATPase, is the responsible gene in more than half of the Zellweger syndrome spectrum patients, and mutations in PEX1 can account for the full spectrum of phenotypes seen in these patients. In these studies, we have undertaken mutation analysis of PEX1 in skin fibroblast cell lines from Australasian Zellweger syndrome spectrum patients. A previously reported common PEX1 mutation that gives rise to a G843D substitution and correlates with the less severe disease phenotypes has been found to be present at high frequency in our patient cohort. We also report a novel PEX1 mutation that occurs at high frequency in Zellweger syndrome spectrum patients. This mutation produces a frameshift in exon 13, a change that leads to the premature truncation of the PEX1 protein. A Zellweger syndrome patient who was homozygous for this mutation and who survived for less than two months from birth had undetectable levels of PEX1 mRNA. This new common mutation therefore correlates with a severe disease phenotype. We have adopted procedures for the detection of this mutation for successful prenatal diagnosis. PMID- 10480354 TI - Mutation analysis of hereditary multiple exostoses in the Chinese. AB - Hereditary multiple exostoses (EXT; MIM 133700) is an autosomal dominant bone disorder. It is genetically heterogeneous with at least three chromosomal loci: EXT1 on 8q24.1, EXT2 on 11p11, and EXT3 on 19p. EXT1 and EXT2, the two genes responsible for EXT1 and EXT2, respectively, have been cloned. Recently, three other members of the EXT gene family, named the EXT-like genes (EXTL: EXTL1, EXTL2, and EXTL3), have been isolated. EXT1, EXT2, and the three EXTLs are homologous with one another. We have identified the intron-exon boundaries of EXTL1 and EXTL3 and analyzed EXT1, EXT2, EXTL1, and EXTL3, in 36 Chinese families with EXT, to identify underlying disease-related mutations in the Chinese population. Of the 36 families, five and 12 family groups have mutations in EXT1 and EXT2, respectively. No disease-related mutation has been found in either EXTL1 or EXTL2, although one polymorphism has been detected in EXTL1. Of the 15 different mutations (three families share a common mutation in EXT2), 12 are novel. Most of the mutations are either frameshift or nonsense mutations (12/15). These mutations lead directly or indirectly to premature stop codons, and the mutations generate truncated proteins. This finding is consistent with the hypothesis that the development of EXT is mainly attributable to loss of gene function. Missense mutations are rare in our families, but these mutations may reflect some functionally crucial regions of these proteins. EXT1 is the most frequent single cause of EXT in the Caucasian population in Europe and North America. It accounts for about 40% of cases of EXT. Our study of 36 EXT Chinese families has found that EXT1 seems much less common in the Chinese population, although the frequency of the EXT2 mutation is similar in the Caucasian and Chinese populations. Our findings suggest a possibly different genetic spectrum of this disease in different populations. PMID- 10480355 TI - Transcript identification on the CLN5 region on chromosome 13q22. AB - Our efforts to clone the CLN5 gene, mutated in a severe children's brain disease, variant late infantile neuronal ceroid lipofuscinosis (vLINCL, MIM256731), resulted in large-scale sequencing of genomic clones flanking the critical chromosomal region on 13q22. Computational and traditional transcript identification analyses of the resulting sequence were used to identify the disease gene. In addition to the identification of the CLN5 gene, this effort produced a large amount of genomic sequence data. Here, we report a transcription map of the 107 kb sequence in the CLN5 region, based on traditional and computational gene identification strategies. Several transcripts were identified in this sequence. Queries against the database of expressed sequence tags proved to be the most powerful tool for gene identification from large-scale sequence. PMID- 10480356 TI - Identification of novel RPGR (retinitis pigmentosa GTPase regulator) mutations in a subset of X-linked retinitis pigmentosa families segregating with the RP3 locus. AB - The X-linked form of retinitis pigmentosa (XLRP) is a severe disease of the retina, characterised by night blindness and visual field constriction in a degenerative process, culminating with complete loss of sight within the third decade of life. Genetic mapping studies have identified two major loci for XLRP: RP3 (70%-75% of XLRP) and RP2 (20%-25% of XLRP). The RPGR (retinitis pigmentosa GTPase regulator) gene has been cloned within the RP3 genomic interval and it has been shown that 10%-20% of XLRP families have mutations in this gene. Here, we describe a single-strand conformational polymorphism-based mutation screening of RPGR in a pool of 29 XLRP families for which the disease segregates with the RP3 locus, in order to investigate the proportion of RP3 families with RPGR mutations and to relate the results to previous reports. Five different new mutations have been identified: two splice site mutations for exon 1 and three frameshift mutations in exons 7, 10 and 11. The percentage of RPGR mutations identified is 17% (5/29) in our genetically well-defined population. This figure is comparable to the percentage of RP2 gene mutations that we have detected in our entire XLRP patient pool (10%-15%). A correlation of RPGR mutations with phenotype in the families described in this study and the biochemical characterisation of reported mutations may provide insights into the function of the protein. PMID- 10480357 TI - Genetic variation in the apolipoprotein H (beta2-glycoprotein I) gene affects plasma apolipoprotein H concentrations. AB - Apolipoprotein H (apoH, protein; APOH, gene) is a single chain glycoprotein that exists in plasma both in a free form and in combination with lipoprotein particles. ApoH has been implicated in several physiologic pathways, including lipid metabolism, coagulation, and the production of antiphospholipid antibodies. The wide range of interindividual variation in plasma apoH levels is thought to be under genetic control, but its molecular basis is unknown. APOH displays a common structural polymorphism with the occurrence of three common alleles (APOH*1, APOH*2, and APOH*3), the APOH*2 allele being the most frequent in all populations. The relationship between the APOH polymorphism and plasma apoH levels is unknown. In this study, we have determined the impact of this APOH polymorphism on apoH levels in 455 normoglycemic non-Hispanic Whites (220 men and 235 women) from the San Luis Valley, Colorado. Mean plasma apoH levels, determined by capture enzyme-linked immunosorbent assay, were 20.0 +/- 0.2 mg/dl (range: 3.4-31.2 mg/dl) with no significant difference between men and women. In women, but not in men, age had a significant effect on plasma apoH levels explaining 3.4% of its phenotypic variance. ApoH levels also correlated positively with cholesterol (P = 0.015), HDL-cholesterol (P = 0.044), and triglyceride (P = 0.037) in women, but not in men. An analysis of variance (ANOVA) of adjusted plasma apoH levels showed significant association with the APOH polymorphism in both men and women (P < 0.0001), and the APOH polymorphism accounted for 11.4% and 13.6% of the variation in apoH levels in men and women, respectively. Compared with the APOH*1 and APOH*2 alleles, the APOH*3 allele was associated with significantly lower plasma apoH levels. At the molecular level, APOH*3 can be further subdivided into two distinct forms, called APOH*3W and APOH*3B. The APOH*3W form is more common in US Whites and is the result of a missense mutation at codon 316. An ANOVA for the codon 316 polymorphism revealed that this polymorphism is a major determinant of plasma apoH variation (P < 0.0001). This study indicates that common genetic variation in the APOH gene is a significant determinant of plasma apoH levels in non-Hispanics Whites and should be useful in evaluating the role of the APOH genetic variation in various metabolic pathways in which apoH has been implicated. PMID- 10480358 TI - A comparison of BRCA1 mutation analysis by direct sequencing, SSCP and DHPLC. AB - The most sensitive screening technique for genes that predispose patients for particular cancers is direct sequencing. However, sequencing of complex genes is technically demanding, costly and time-consuming. We have tested alternate screening techniques to find a fast sensitive method for detecting alterations of DNA in the large BRCA1 gene prior to sequencing. Sequencing of this gene is particularly arduous because it lacks clearly defined mutation sites. The single strand conformation polymorphism (SSCP) technique is one of the most frequently used pre-screening methods but its sensitivity and efficiency is not completely satisfying. We have compared the SSCP assay with a newly developed technique called denaturing high performance liquid chromatography (DHPLC) to screen the BRCAl gene. We studied 23 patients at high risk for early onset breast and ovarian cancer and four controls. In these patients, a total of 113 fragments with sequence variations in the BRCA1 gene could be identified. The DHPLC technique resolved 100% of the DNA alterations that were observed in cycle sequencing. In contrast, mutation analysis by SSCP accounted for 94% of the detected variations. In addition, DHPLC screening allowed us to discriminate between different alterations in a single fragment, because of the characteristic elution profiles of the DNA molecules. Polymorphisms that were present in our samples could be predicted by means of DHPLC testing independently of sequence analysis. We conclude that DHPLC is a highly potent screening method for genetic analyses. It is highly sensitive, efficient and economical and can be automated. PMID- 10480360 TI - FISH-detected delay in replication timing of mutated FMR1 alleles on both active and inactive X-chromosomes. AB - X-chromosome inactivation and the size of the CGG repeat number are assumed to play a role in the clinical, physical, and behavioral phenotype of female carriers of a mutated FMR1 allele. In view of the tight relationship between replication timing and the expression of a given DNA sequence, we have examined the replication timing of FMR1 alleles on active and inactive X-chromosomes in cell samples (lymphocytes or amniocytes) of 25 females: 17 heterozygous for a mutated FMR1 allele with a trinucleotide repeat number varying from 58 to a few hundred, and eight homozygous for a wild-type allele. We have applied two-color fluorescence in situ hybridization (FISH) with FMR1 and X-chromosome alpha satellite probes to interphase cells of the various genotypes: the alpha satellite probe was used to distinguish between early replicating (active) and late replicating (inactive) X-chromosomes, and the FMR1 probe revealed the replication pattern of this locus. All samples, except one with a large trinucleotide expansion, showed an early replicating FMR1 allele on the active X chromosome and a late replicating allele on the inactive X-chromosome. In samples of mutation carriers, both the early and the late alleles showed delayed replication compared with normal alleles, regardless of repeat size. We conclude therefore that: (1) the FMR1 locus is subjected to X-inactivation; (2) mutated FMR1 alleles, regardless of repeat size, replicate later than wild-type alleles on both the active and inactive X-chromosomes; and (3) the delaying effect of the trinucleotide expansion, even with a low repeat size, is superimposed on the delay in replication associated with X-inactivation. PMID- 10480359 TI - Prevalence of germline mutations of hMLH1, hMSH2, hPMS1, hPMS2, and hMSH6 genes in 75 French kindreds with nonpolyposis colorectal cancer. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is a syndrome characterized by familial predisposition to colorectal carcinoma and extracolonic cancers of the gastrointestinal, urological, and female reproductive tracts. This dominant disorder is caused by germline defects in one of at least five DNA mismatch repair (MMR) genes: hMLH1, hMSH2, hPMS1, hPMS2, and hMSH6 (GTBP). Germline mutations of hMSH2 and hMLH1 are also frequently identified in families not fulfilling all the Amsterdam criteria, thereby demonstrating that the involvement of these genes is not confined to typical HNPCC. To evaluate the respective involvement of the various MMR genes in typical and incomplete HNPCC syndromes, we have performed an analysis of the hMLH1, hMSH2, hPMS1, hPMS2, and hMSH6 genes in a large series of French kindreds (n=75) with colorectal tumors and/or aggregation of extracolonic cancers belonging to the HNPCC spectrum. Mutational analysis has been performed in all families, without preselection for the tumor phenotype. We have detected 26 pathogenic germline mutations of the hMLH1 and hMSH2 genes and several novel variants of the hPMS1, hPMS2, and hMSH6 genes. Our data confirm that, regardless of the type of families and the tumor phenotype, hPMS1, hPMS2, and hMSH6 germline mutations are rare in familial aggregation of colorectal cancers. Furthermore, they suggest that the presence of multiple primary malignancies in a single individual and the observation of extracolonic tumors in relatives of a colorectal cancer patient should be included among the guidelines for referring patients for genetic testing. PMID- 10480361 TI - The human CDC42 gene: genomic organization, evidence for the existence of a putative pseudogene and exclusion as a SJS1 candidate gene. AB - Schwartz-Jampel syndrome (SJS) is an autosomal recessive human disorder characterized by myotonia and osteoarticular deformities. Three types are distinguished based on age at onset: types 1A, 1B and 2. We have previously localized the SJS1 gene, responsible for types 1A and 1B, on human chromosome 1p35-p36.1 in a region frequently rearranged in human tumours. The CDC42 gene, for which divergent localizations have previously been described (chromosomes 4, 7 and 20), has been mapped within the SJS1 critical interval by radiation hybrid and yeast/P1 artificial-chromosome-based physical map analyses. The CDC42 gene product is a small GTPase protein of the Rho family mediating a variety of signaling pathways including cytoskeletal rearrangements, cell-cycle progression and transformation. To search for mutations in SJS1 patients, we have determined the organization of the human CDC42 gene on chromosome 1p and found that it encodes for the placental and brain isoforms generated by alternative splicing. No mutations have been found in SJS1 patients, excluding CDC42 as the SJS1 gene. Interestingly, we have demonstrated that a CDC42-like transcript gene located on chromosome 4 does not contain introns and is similar to the placental isoform, suggesting that it is a processed pseudogene. The determination of the CDC42 gene structure described in this report should facilitate future studies of the potential role of CDC42 in human disorders. PMID- 10480362 TI - Genomic organisation of the human chordin gene and mutation screening of candidate Cornelia de Lange syndrome genes. AB - We have determined the genomic organisation of the human chordin gene, CHRD, and have shown that it maps within a gene cluster at 3q27 containing THPO (thrombopoietin), CLCN2 (a voltage-gated chloride-channel gene) and EIF4G1 (a eukaryotic translation-initiation-factor-gamma gene). The CHRD and THPO genes are very close neighbours and are transcribed from opposing DNA strands from promoters that are spaced less than 2 kb apart. We considered that the CHRD gene and the chordin-regulating GSC (goosecoid) gene could be candidate genes for Cornelia de Lange syndrome (CDLS), a developmental malformation syndrome which is primarily characterised by mental handicap, growth retardation, distinctive facial features and limb-reduction defects. CDLS patients typically occur as sporadic cases, but several reports have suggested dominant inheritance. The candidacy of the CHRD and GSC genes was supported by several lines of evidence: prior evidence for a CDLS gene at 3q26.3-q27; a report suggesting a significant association between CDLS and thrombocytopenia; suspected genetic heterogeneity in CDLS; location of the GSC gene in close proximity to a 14q32 breakpoint detected in a CDLS patient with a balanced de novo translocation; known regulation of chordin expression by goosecoid; and the pattern of embryonic expression of the mouse GSC gene. Another candidate gene at 3q27, SOX2, was also considered because of its suspected role as a transcription factor in early development and because of known examples of SOX genes that are loci for dominantly inherited developmental disorders. However, mutation screening failed to identify CDLS patient-specific mutations in CHRD, GSC or SOX2. PMID- 10480363 TI - A familial case of recurrent hydatidiform molar pregnancies with biparental genomic contribution. AB - Hydatidiform mole is a benign trophoblastic neoplasia characterized by an abnormal development of the embryo and proliferation of placental villi. Using microsatellite markers amplified by the polymerase chain reaction, we have performed a genetic study on eight independent molar tissues occurring in two sisters. Karyotype and genotype data demonstrate a diploid and biparental constitution in seven of the analyzed moles suggesting a common mechanism underlying the etiology of the various molar pregnancies in this family. The data reported here suggest that complete and partial hydatidiform moles are not always separate entities and that women with familial recurrent hydatidiform moles are homozygous for an autosomal recessive mutation. PMID- 10480364 TI - X-linked adrenomyeloneuropathy associated with 14 novel ALD-gene mutations: no correlation between type of mutation and age of onset. AB - Adrenomyeloneuropathy (AMN) represents a milder form of X-linked adrenoleukodystrophy (ALD), the most frequent peroxisomal disorder. The disease is characterised by an abnormal accumulation of saturated, very long chain, fatty acids, because of altered peroxisomal beta-oxidation that concomitantly leads to demyelination in the central and peripheral nervous systems. ALD shows a highly variable phenotypic expression and extensive mutation analysis in ALD patients has failed to establish a genotype-phenotype correlation, even in the presence of the same ALD-gene defect. Therefore, we have looked for a relationship between the molecular lesion and the age of onset in 19 patients with a well-classified clinical course of AMN. The nearly complete novel spectrum of ALD gene mutations identified has revealed no obvious correlation between the type of mutation and age of AMN onset in this small series. However, intrafamiliar concordance could be observed with respect to the occurrence of adrenocortical insufficiency. This supports the idea of one (or more) additional gene(s) contributing to the phenotypic expression of ALD. PMID- 10480365 TI - Clinical and genetic studies of Japanese homozygotes for the Gaucher disease L444P mutation. AB - In patients originally genotyped as homoallelic for the Gaucher disease (GD) L444P (1448C) mutation, we sought to confirm previously reported phenotypic differences between Caucasians and Japanese, to determine the prevalence and phenotypic impact of recombinant alleles, and to explore the phenotypic influence of genetic background. We therefore analyzed data from longer-term clinical follow-up, more comprehensive genotyping and polymorphism and mitochondrial DNA (mtDNA) testing in all known Japanese L444P homozygotes (n=15). Our studies demonstrated that, of 12 patients in our series originally diagnosed with non neuronopathic GD, 9 developed neurological signs/symptoms during follow-up (at a mean of 14 years 11 months +/- 11 years 4 months). Of three patients originally diagnosed with acute neuronopathic (type 2) GD, all three were compound heterozygotes for L444P and the complex allele RecNci I. In the entire series, Pvu II and liver erythrocyte pyruvate kinase (PKLR) polymorphism and prevalence of the 9 bp mtDNA deletion were heterogeneous, and these background genetic factors could not predict phenotypic expression. Our data suggest that, in Japanese as in Caucasian patients, the L444P/L444P genotype is highly associated with subacute neuronopathic (type 3) GD, and the presence of a complex allele together with an L444P allele leads to type 2 disease. Our findings also underline the importance of comprehensive genotyping (particularly testing for recombinant alleles), long-term follow-up and careful neurological examination in patients with early-onset GD. Such measures ultimately may improve genotype/phenotype correlations and, with them, genetic counseling and therapeutic decision making. PMID- 10480366 TI - A unique 3.5-kb deletion of the mitochondrial genome in Thai patients with Kearns Sayre syndrome. AB - Kearns-Sayre syndrome is one of the neurological diseases caused by a defect in the energy-producing system of mitochondria. Keams-Sayre is known to be associated with a deletion in the mitochondrial genome and is usually detected in muscle biopsies of the patients. In this study, we report the molecular lesion of mitochondrial DNA (mtDNA) in four Thai patients admitted to hospital with encephalomyopathies. The 3.5-kb deletion of mtDNA was detected by Southern analysis, mapped by amplification with five primer pairs covering almost the total mitochondrial genome, and confirmed by PCR primer shift analysis. The deleted position was localized to nt 10208/13765 or nt 10204/13761 spanning the coding area of subunits 3 (ND3), 4L (ND4L), 4 (ND4), and 5 (ND5) of respiratory chain enzyme complex I and the tRNA genes for histidine, serine, leucine, and arginine. The sequence flanking the deletion was a 4-bp repeat of TCCC. All four patients have exactly the same 3558-bp mtDNA deletion; this is the only deleted position in their mtDNA but is different from those reported in the literature. The deletion seems to be found only in Thai patients, although they present with different clinical manifestations and none of them is not related. PMID- 10480367 TI - Cell fusion corrects the 4-nitroquinoline 1-oxide sensitivity of Werner syndrome fibroblast cell lines. AB - We have shown that Werner syndrome (WRN) fibroblast cell lines are unusually sensitive to the DNA-damaging agent 4-nitroquinoline 1-oxide (4NQO), though not to gamma radiation or to hydrogen peroxide. The fusion of 4NQO-sensitive WRN and 4NQO-resistant control fibroblast cell lines generated proliferating WRN x control cell hybrids that expressed WRN protein and were 4NQO-resistant. These results establish the recessive nature of 4NQO sensitivity in WRN cell lines and provide a cellular assay for WRN protein function. PMID- 10480368 TI - Genomic structure and expression analysis of the spastic paraplegia gene, SPG7. AB - SPG7 is a newly identified gene involved in an autosomal recessive form of hereditary spastic paraplegia (HSP), a genetically heterogeneous group of neurodegenerative disorders. This gene encodes a protein characterized as a nuclear-encoded mitochondrial metalloprotease. The present report describes the genomic structure of the SPG7 gene. It is organized into 17 exons ranging from 78 to 242 bp and spans approximately 52 kb within three overlapping cosmids. The exon/intron boundaries and all splice junctions are consistent with the published consensus sequences for donor and acceptor sites. The provided genomic structure of SPG7 should facilitate the screening for mutations in this gene in patients with HSP and other related mitochondrial disease syndromes. SPG7 has been mapped to chromosome 16q24.3, a region of frequent loss of heterozygosity (LOH) seen in sporadic breast and prostate cancer. We have performed single-strand conformation polymorphism analysis of ten exons of this gene in a number of sporadic breast cancer samples showing LOH at 16q24.3. No mutations were detected; only single nucleotide polymorphisms were observed in exon 11, intron 7, intron 10 and intron 12. An expression analysis study has revealed the differential expression of SPG7 mRNA in various tissues and at different developmental stages. PMID- 10480369 TI - Complex allele [-102T>A+S549R(T>G)] is associated with milder forms of cystic fibrosis than allele S549R(T>G) alone. AB - We recently reported a novel complex allele in the cystic fibrosis transmembrane regulator (CFTR) gene, combining a sequence change in the minimal CFTR promoter ( 102T>A) and a missense mutation in exon 11 [S549R(T>G)]. Here we compare the main clinical features of six patients with cystic fibrosis (CF) carrying the complex allele [-102T>A+S549R(T>G)] with those of 16 CF patients homozygous for mutation S549R(T>G) alone. Age at diagnosis was higher, and current age was significantly higher (P=0.0032) in the group with the complex allele, compared with the S549R/S549R group. Although the proportion of patients with lung colonization was similar in both groups, the age at onset was significantly higher in the group with the complex allele (P=0.0022). Patients with the complex allele also had significantly lower sweat test chloride values (P=0.0028) and better overall clinical scores (P=0.004). None of the 22 patients reported in this study had meconium ileus. All 16 patients homozygous for S549R(T>G), however, were pancreatic insufficient, as compared with 50% of patients carrying the complex allele (P=0.013). Moreover, the unique patient homozygous for [ 102T>A+S549R(T>G)] presented with a mild disease at 34 years of age. These observations strongly suggest that the sequence change (-102T>A) in the CFTR minimal promoter could attenuate the severe clinical phenotype associated with mutation S549R(T>G). PMID- 10480370 TI - The frequency of lysosomal storage diseases in The Netherlands. AB - We have calculated the relative frequency and the birth prevalence of lysosomal storage diseases (LSDs) in The Netherlands based on all 963 enzymatically confirmed cases diagnosed during the period 1970-1996. The combined birth prevalence for all LSDs is 14 per 100,000 live births. Glycogenosis type II is the most frequent LSD with a birth prevalence of 2.0 per 100,000 live births, representing 17% of all diagnosed cases. Within the group of lipidoses, metachromatic leukodystrophy (MLD) is the most frequent LSD. MLD was diagnosed in 24% of lipidoses and the calculated birth prevalence was 1.42 per 100,000 for all types combined. Krabbe disease, diagnosed in 17% of cases, also belongs to the more frequent lipid storage diseases in The Netherlands with a birth prevalence of 1.35 per 100,000. The birth prevalence of Gaucher disease, commonly regarded as the most frequent lipid storage disease is 1.16 per 100,000 for all types combined. The combined birth prevalence for all lipid storage diseases is 6.2 per 100,000 live births. Within the group of mucopolysaccharidoses (MPSs), MPS I has the highest calculated birth prevalence of 1.19 per 100,000 (25% of all cases of MPS diagnosed), which is slightly more frequent than MPS IIIA with an estimated birth prevalence of 1.16 per 100,000. As a group, MPS III comprises 47% of all MPS cases diagnosed and the combined birth prevalence is 1.89 per 100,000 live births. The birth prevalence of MPS II is 0.67 per 100,000 (1.30 per 100,000 male live births). All other MPSs are rare. The combined birth prevalence for all MPSs is 4.5 per 100,000 live births. Mucolipidoses and oligosaccharidoses are very rare with birth prevalences between 0.04 and 0.20 for individual diseases. Only 49 cases were diagnosed between 1970 and 1996. Their combined birth prevalence is 1.0 per 100,000 live births. PMID- 10480371 TI - Identification of two novel mutations in OCTN2 of three patients with systemic carnitine deficiency. AB - Systemic carnitine deficiency is a potentially lethal, autosomal recessive disorder characterized by cardiomyopathy, myopathy, recurrent episodes of hypoketotic hypoglycemia, hyperammonemia, and failure to thrive. This form of carnitine deficiency is caused by a defect in the active cellular uptake of carnitine, and the gene encoding the high affinity carnitine transporter OCTN2 has recently been shown to be mutated in patients suffering from this disorder. Here, we report the underlying molecular defect in three unrelated patients. Two patients were homozygous for the same missense mutation 632A-->G, which changes the tyrosine at amino acid position 211 into a cysteine (Y211C). The third patient was homozygous for a nonsense mutation, 844C-->T, which converts the arginine at amino acid position 282 into a stop codon (R282X). Reintroduction of wild-type OCTN2 cDNA into fibroblasts of the three patients by transient transfection restored the cellular carnitine uptake, confirming that mutations in OCTN2 are the cause of systemic carnitine deficiency. PMID- 10480372 TI - Physical mapping of the G-protein coupled receptor 19 (GPR19) in the chromosome 12p12.3 region frequently rearranged in cancer cells. AB - Rearrangements of the short arm of chromosome 12 are frequently observed in hematological malignancies and in certain solid neoplasias. Loss of heterozygosity studies identified a small genetic interval on chromosome 12p12.3 that is frequently deleted in childhood acute lymphoblastic leukemia (ALL). Two genes, ETV6 and CDKN1B, are located within this interval although evidence has accumulated that an as yet unidentified tumor suppressor gene is closely linked. Here we report the physical mapping of the G-protein coupled receptor 19 (GPR19) at approximately 40kb from CDKN1B. The delineation of the gene order tel-ETV6 CDKN1B-GPR19-cen excluded GPR19 from the region commonly deleted in childhood ALL, but it could still be the target of genetic alterations found in other cancers. PMID- 10480373 TI - CTG repeats distribution and Alu insertion polymorphism at myotonic dystrophy (DM) gene in Amhara and Oromo populations of Ethiopia. AB - Myotonic dystrophy (DM) is a dominantly inherited neuromuscular disease, highly variable and multisystemic, which is caused by the expansion of a CTG repeat located in the 3' untranslated region of the DMPK gene. Normal alleles show a copy number of 5-37 repeats on normal chromosomes, amplified to 50-3000 copies on DM chromosomes. The trinucleotide repeat shows a trimodal allele distribution in the majority of the examined population. The first class includes alleles carrying (CTG)5, the second class, alleles in the range 7-18 repeats, and the third class, alleles (CTG) > or =19. The frequency of this third class is directly related to the prevalence of DM in different populations, suggesting that normal large-sized alleles predispose toward DM. We studied CTG repeat allele distribution and Alu insertion and/or deletion polymorphism at the myotonic dystrophy locus in two major Ethiopian populations, the Amhara and Oromo. CTG allele distribution and haplotype analysis on a total of 224 normal chromosomes showed significant differences between the two ethnic groups. These differences have a bearing on the out-of-Africa hypothesis for the origin of the DM mutation. In addition, (CTG) > or =19 were exclusively detected in the Amhara population, confirming the predisposing role of these alleles compared with the DM expansion-mutation. PMID- 10480374 TI - Connexin 50 mutation in a family with congenital "zonular nuclear" pulverulent cataract of Pakistani origin. AB - Inherited cataract is a clinically and genetically heterogeneous disease that most often presents as a congenital autosomal dominant trait. Here we report linkage of a three-generation family of Pakistani origin with autosomal dominant cataract "zonular nuclear" pulverulent type (CZNP) on chromosome 1q21.1. Genome wide-linkage analysis excluded all the known cataract loci except on chromosome 1q. Significantly positive 2-point lod score values (Z=3.01 at theta=0) were obtained for markers D1S305 and D1S2721, which are known to flank the gene for connexin 50 (Cx50) or gap junction protein alpha-8 (Gja8). Previously a mutation in this gene has been reported in a British family with zonular pulverulent cataract (CZP). Here we describe a second mutation (E48K) in connexin 50 that confirms the involvement of this gene in cataractogenesis. PMID- 10480375 TI - A new hereditary cylindromatosis family associated with CYLD1 on chromosome 16. AB - Hereditary cylindromatosis (HC; MIM 132700) is an autosomal dominant condition characterized by benign skin appendage tumors most commonly on the scalp and face. Previously, the HC gene (CYLD1) was linked to chromosome 16q12-13, and tumors showed loss of heterozygosity (LOH), suggesting that CYLD1 is a tumor suppressor gene. Here we report a new multi-generation cylindromatosis family whose condition maps to that region, with 7/13 tumors showing LOH on 16q. PMID- 10480376 TI - The genomic structure and developmental expression patterns of the human OPA containing gene (HOPA). AB - We determined the genomic organization of the human OPA-containing gene (HOPA) and characterized its developmental expression. The gene encoding HOPA, which contains a rare polymorphism tightly associated with non-specific mental retardation, is 25 kb in length and consists of 44 exons. A promoter scan analysis demonstrates two possible transcription initiation sites without TATA boxes upstream from the putative translation initiation start site. Several informative polymorphisms are evident in the sequence including a large pentanucleotide repeat. Northern blot analysis of the gene transcript and its murine orthologue, MOPA-1, demonstrates that only one transcript is expressed throughout the soma and the CNS, and that the transcript is highly expressed during early fetal development. We conclude that the delineation of the function of the HOPA gene locus merits further study. PMID- 10480377 TI - Association between M/L55-polymorphism of paraoxonase enzyme and oxidative DNA damage in patients with type 2 diabetes mellitus and in control subjects. AB - The paraoxonase enzyme (PON) gene polymorphism causes a change of methionine (M allele) to leucine (L-allele). PON may reduce low density lipoprotein oxidation and prevent atherosclerosis. Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive index of oxidative DNA damage. We have studied the association between the PON genotypes and the urinary excretion of 8-OHdG. The study population consisted of 93 Finnish type 2 diabetes patients and 106 non-diabetic control subjects. The 24-h excretion of 8-OHdG was significantly higher in diabetic patients than in control subjects (P < 0.001). In control subjects, the ratio of the 8-OHdG/glomerular filtration rate increased in order of genotype from MM to ML to LL (P < 0.0412). These results suggest that lipid peroxidation may have an effect on DNA oxidation. PMID- 10480378 TI - Chromosome 9qh inversions may not be true inversions. PMID- 10480379 TI - 14-3-3 proteins: eukaryotic regulatory proteins with many functions. AB - The enigmatically named 14-3-3 proteins have been the subject of considerable attention in recent years since they have been implicated in the regulation of diverse physiological processes, in eukaryotes ranging from slime moulds to higher plants. In plants they have roles in the regulation of the plasma membrane H+-ATPase and nitrate reductase, among others. Regulation of target proteins is achieved through binding of 14-3-3 to short, often phosphorylated motifs in the target, resulting either in its activation (e.g. H+-ATPase), inactivation (e.g. nitrate reductase) or translocation (although this function of 14-3-3 proteins has yet to be demonstrated in plants). The native 14-3-3 proteins are homo- or heterodimers and, as each monomer has a binding site, a dimer can potentially bind two targets, promoting their association. Alternatively, target proteins may have more than one 14-3-3-binding site. In this mini review, we present a synthesis of recent results from plant 14-3-3 research and, with reference to known 14-3-3-binding motifs, suggest further subjects for research. PMID- 10480380 TI - Secondary xylem-specific expression of caffeoyl-coenzyme A 3-O-methyltransferase plays an important role in the methylation pathway associated with lignin biosynthesis in loblolly pine. AB - Two types of structurally distinct O-methyltransferases mediate the methylation of hydroxylated monomeric lignin precursors in angiosperms. Caffeate 3-O methyltransferase (COMT; EC 2.1.1.68) methylates the free acids and caffeoyl CoA 3-O-methyltransferase (CCoAOMT; EC 2.1.1.104) methylates coenzyme A esters. Recently, we reported a novel hydroxycinnamic acid/hydroxycinnamoyl CoA ester O methyltransferase (AEOMT) from loblolly pine differentiating xylem that was capable of methylating both acid and ester precursors with similar efficiency. In order to determine the possible existence and role of CCoAOMT in lignin biosynthesis in gymnosperms, a 1.3 kb CCoAOMT cDNA was isolated from loblolly pine that showed 79-82% amino acid sequence identity with many angiosperm CCoAOMTs. The recombinant CCoAOMT expressed in Escherichia coli exhibited a significant methylating activity with hydroxycinnamoyl CoA esters whereas activity with hydroxycinnamic acids was insignificant. Moreover, 3.2 times higher catalytic efficiency for methylating caffeoyl CoA over 5-hydroxyferuloyl CoA was observed which could serve as a driving force towards synthesis of guaiacyl lignin. The secondary xylem-specific expression of CCoAOMT was demonstrated using RNA blot analysis, western blot analysis, and O-methyltransferase enzyme assays. In addition, Southern blot analysis indicated that CCoAOMT may exist as a single copy gene in loblolly pine genome. The transgenic tobacco plants carrying loblolly pine CCoAOMT promoter-GUS fusion localized the site of GUS activity at the secondary xylem tissues. These data suggest that CCoAOMT, in addition to AEOMT, plays an important role in the methylation pathway associated with lignin biosynthesis in loblolly pine. PMID- 10480381 TI - Identification and characterization of T3/T7 bacteriophage-like RNA polymerase sequences in wheat. AB - Using PCR-based methods, we assembled two wheat cDNA sequences, wheat-G and wheat C, that encode T3/T7 bacteriophage-like RNA polymerases (RNAPs) sharing 45% amino acid identity. In phylogenetic analyses using maximum likelihood, parsimony and distance methods, the predicted protein sequence of wheat-G (1005 amino acids, 113 kDa) clusters with sequences of previously assigned mitochondrial RNAPs from dicotyledonous plants (Arabidopsis thaliana, Chenopodium album); likewise, in such analyses, the wheat-C sequence (949 amino acids, 107 kDa) affiliates specifically with the Arabidopsis sequence that encodes a phage-like RNAP thought to function in chloroplasts. To confirm biochemically the assignment of the gene encoding the putative wheat mitochondrial RNAP, we isolated a ca. 100 kDa wheat mitochondrial protein that is enriched in fractions displaying specific in vitro transcription activity and that reacts with an antibody raised against a recombinant maize phage-type RNAP. Internal peptide sequence information obtained from the 100-kDa polypeptide revealed that it corresponds to the predicted wheat G cDNA sequence, providing direct evidence that the wheat-G gene (which we propose to call RpoTm) encodes the wheat mitochondrial RNAP. PMID- 10480382 TI - Early elicitor induction in members of a novel multigene family coding for highly related RING-H2 proteins in Arabidopsis thaliana. AB - We describe the identification and structural characterization of a novel family of Arabidopsis genes related to ATL2 which encode a variant of the RING zinc finger domain, known as RING-H2. Analysis of genes selected by us and of sequences from Arabidopsis stored in databases permitted the prediction of several RING-H2 proteins that contain highly homologous RING domains. The ATL gene family is represented by fifteen sequences that contain, in addition to the RING, a transmembrane domain which is located in most of them towards the N terminal end. Transgenic Arabidopsis seedlings carrying the ATL2 promoter fused to the GUS reporter gene revealed that the expression of ATL2 is rapidly induced after exposure to chitin or inactivated crude cellulase preparations. Rapid induction of transcript accumulation of another member of the ATL family was also observed under the same conditions. These results suggest that some ATLs may be involved in the early stages of the defense response triggered in plants in response to pathogen attack. PMID- 10480383 TI - An anaerobically inducible early (aie) gene family from rice. AB - One of the major abiotic stresses that affect plant growth and development is anoxia or hypoxia. Plants respond to anoxia by regulation of gene expression at both the transcriptional and translational levels. Genes involved in such regulation are expected to be expressed soon after onset of anoxia. To date, however, anaerobically regulated genes that have been characterized predominantly encode enzymes for sugar phosphate metabolism, and are induced after several hours of anaerobic conditions. Early induced genes, those responding after 1-2 h of anoxia, have not been studied extensively. To study the early anaerobic response we investigated the most flooding-tolerant variety of rice, FR13A (flood resistant). We used differential display techniques to identify cDNA fragments representing mRNAs that are induced within 90 min of anoxia. We isolated two cDNA fragments and one full-length cDNA that were induced to high levels. These cDNAs were found to be members of a family of 2-3 genes, which were called the aie (anaerobically inducible early) gene family. Northern blot analyses showed that the mRNA levels of aie genes peaked after 1.5 to 3 h of anoxia and were still at high levels after 72 h of anoxia. RNase protection assays showed 4-5 different protected bands indicating multiple transcripts from the aie gene family. Sequence analyses of the full-length cDNA showed an open reading frame that putatively encodes a 14 kDa protein of 127 amino acid residues. Neither the nucleotide nor the deduced amino acid sequences of this gene showed any significant homology to any known genes or proteins present in the GenBank or SwissProt databases. This novel gene, that is induced so early under anoxia in plants, may play an important role in plant metabolism under anaerobic conditions. PMID- 10480384 TI - A defective seed coat pattern (Net) is correlated with the post-transcriptional abundance of soluble proline-rich cell wall proteins. AB - The pigmented seed coats of several soybean (Glycine max (L.) Merr.) plant introductions and isolines have unusual defects that result in cracking of the mature seed coat exposing the endosperm and cotyledons. It has previously been shown that the T (tawny) locus that controls the color of trichomes on stems and leaves also has an effect on both the structure and pigmentation of the seed coat. Distribution of pigmentation on the seed coat is controlled by alleles of the I (inhibitor) locus. It was also found that total seed coat proteins were difficult to extract from pigmented seed coats with i T genotypes because they have procyanidins that exhibit tannin properties. We report that the inclusion of poly-L-proline in the extraction buffer out-competes proteins for binding to procyanidins. Once this problem was solved, we examined expression of the proline rich cell wall proteins PRP1 and PRP2 in pigmented genotypes with the dominant T allele. We found that both homozygous i T and i t genotypes have reduced soluble PRP1 levels. The epistatic interaction of the double recessive genotype at both loci is necessary to produce the pigmented, defective seed coat phenotype characteristic of seed coats with the double recessive i and t alleles. This implies a novel effect of an enzyme in the flavonoid pathway on seed coat structure in addition to its effect on flavonoids, anthocyanidins, and proanthocyanidins. No soluble PRP1 polypeptides were detectable in pigmented seed coats (i T genotypes) of isolines that also display a net-like pattern of seed coat cracking, known as the Net defect. PRP2 was also absent in one of the these lines. However, both PRP1 and PRP2 cytoplasmic mRNAs were found in the Net defective seed coats. Together with in vitro translation studies, these results suggest that the absence of soluble PRP polypeptides in the defective Net lines is post-translational and could be due to a more rapid or premature insolubilization of PRP polypeptides within the cell wall matrix. PMID- 10480385 TI - Antisense suppression of tomato endo-1,4-beta-glucanase Cel2 mRNA accumulation increases the force required to break fruit abscission zones but does not affect fruit softening. AB - Plants of tomato (Lycopersicon esculentum Mill. cv. T5) were transformed with an antisense endo-1,4-beta-glucanase (cellulase, EC 3.2.1.4) Cel2 transgene under the control of the constitutive cauliflower mosaic virus 35S promoter in order to suppress mRNA accumulation of Cel2. In two independent transgenic lines, Cel2 mRNA abundance was reduced by >95% in ripe fruit pericarp and ca. 80% in fruit abscission zones relative to non-transgenic controls. In both transgenic lines the softening of antisense Cel2 fruit pericarp measured using stress-relaxation analysis was indistinguishable from control fruit. No differences in ethylene evolution were observed between fruit of control and antisense Cel2 genotypes. However, in fruit abscission zones the suppression of Cel2 mRNA accumulation caused a significant (P<0.001) increase in the force required to cause breakage of the abscission zone at 4 days post breaker, an increase of 27% in one transgenic line and of 46% in the other transgenic line. Thus the Cel2 gene product contributes to cell wall disassembly occurring in cell separation during fruit abscission, but its role, if any, in softening or textural changes occurring in fruit pericarp during ripening was not revealed by suppression of Cel2 gene expression. PMID- 10480386 TI - Expression analysis of a cytosolic glutamine synthetase gene in cotyledons of Scots pine seedlings: developmental, light regulation and spatial distribution of specific transcripts. AB - The expression of a cytosolic glutamine synthetase (GS1; EC 6.3.1.2) gene was examined in cotyledons of Scots pine seedlings. Light strongly stimulated GS1 mRNA accumulation during development. Similarly, steady-state levels of GS1 transcripts increased in dark-grown seedlings transferred to light and decreased in dark-adapted seedlings. Light/dark adaptation affected rbcS and lhcb2 mRNA levels and chlorophyll contents in the same manner. Light-grown seedlings in the presence of the herbicide norflurazon showed a drastic decrease in mRNA for GS and photosynthetic proteins, whereas the effect of the herbicide on mitochondrial beta-ATP synthase mRNA was limited. These results indicate that factors associated with developing chloroplasts could be required for maximal GS1 gene expression during seedling development. The level of GS polypeptide, determined by immunoblot, was up-regulated during seedling development in the light or dark. However, the levels of the polypeptide detected were unaltered by the light/dark adaptation treatments. The analysis of GS1 mRNA association with polysomes indicated that the discrepancies between GS protein and mRNA levels are not a result of a differential translational rate of the transcript in darkness relative to light. Two GS isoproteins with different isoelectric point were resolved by two-dimensional PAGE in light- and dark-germinated plants. The relative abundance of one of them was markedly affected by light and correlated with the observed changes in GS mRNA, suggesting that the other form is not a product derived from the detected transcript. In situ hybridization of cotyledon sections showed the presence of GS1 mRNAs in mesophyll and phloem cells confirming gene expression in photosynthetic tissues. High levels of transcript were detected also in meristematic cells of apical primordia. These data suggest a dual role for the GS1 gene associated with chloroplast development/activity and glutamine biosynthesis for nitrogen mobilization during early growth of Scots pine. PMID- 10480387 TI - Characterization and expression of plasma and tonoplast membrane aquaporins in primed seed of Brassica napus during germination under stress conditions. AB - Two aquaporin genes were isolated from a cDNA library of canola (Brassica napus L.). The first aquaporin, BnPIP1 of 1094 bp, encoding a putative polypeptide of 287 amino acids with a predicted molecular mass of 30.4 kDa and a pI of 7.8, belongs to the family of plasma membrane intrinsic protein (PIPs) aquaporins. The B. napus aquaporin showed 85-94% identity to the Arabidopsis thaliana PIPs. ABA priming of seed induced high levels of BnPIP1 transcript which remained after subsequent re-drying of the seed. The second aquaporin, Bny-TIP2 of 1020 bp, encoded a putative polypeptide of 253 amino acids with a predicted molecular mass of 25.8 kDa and a pI of 5.8. Bngamma-TIP2 showed 83-90% identity to gamma-TIP genes from a variety of plant species. Bngamma-TIP2 was expressed only when radicle protrusion occurred in either untreated or primed seeds. Seeds primed with PEG or ABA germinated earlier and showed a higher final percentage of germination than unprimed seed, particularly under salt and osmotic stresses at low temperature. Transcripts of both BnPIP1 and Bngamma-TIP2 genes were present earlier during germination of primed seeds than non-primed seed. From these results, we conclude that BnPIP1 is related to the water transportation required for enzymatic metabolism of storage nutrients at the early stages of canola seed germination whereas Bngamma-TIP2 expression is related to cell growth associated with radicle protrusion. Priming induced the expression of BnPIP1 but had no effect on Bngamma-TIP2. PMID- 10480388 TI - Gibberellin-responsive genes: high level of transcript accumulation in leaf sheath meristematic tissue from Zea mays L. AB - In order to identify genes that are related to the gibberellin (GA) response in maize (Zea mays L.), mRNA species from wild-type and single-gene dwarf mutants, d5 and D8, were compared by fluorescent differential display. The d5 mutant is unable to produce biologically active GA, but recovers its tall stature on exogenous application of GA. D8 is insensitive to GA, despite the accumulation of a high level of endogenous GA, suggesting it to be a receptor mutant or a mutant in signal transduction pathway(s). After screening 7000 cDNA populations, one clone was isolated, for which transcripts were rare in d5 shoots but accumulated within 1 h after GA3 application. This clone, designated as ZmGR1a, encodes a polypeptide with a relative molecular mass of ca. 13 kDa, which shows significant homology to proline-rich proteins from several plant species. A similar experiment with D8 identified a clone, ZmGR2a, with low transcript levels, but accumulation within 6 h after GA3 treatment of d5 shoots. ZmGR2a encodes a polypeptide with a relative molecular mass of ca. 19 kDa, which shows no significant homology with any known protein. Southern blot analysis indicated that ZmGR1a and ZmGR2a form a small multigene family within the maize genome. In situ hybridization with wild-type seedlings showed transcripts on both to be abundant in leaf sheath meristematic tissue, in which GA enhances cell elongation and cell division. PMID- 10480389 TI - Introduction of SLG (S locus glycoprotein) alters the phenotype of endogenous S haplotype, but confers no new S haplotype specificity in Brassica rapa L. AB - Self-incompatibility (SI) in Brassicaceae is genetically controlled by the S locus complex in which S locus glycoprotein (SLG) and S receptor kinase (SRK) genes have been identified, and these two genes encoding stigma proteins are believed to play important roles in SI recognition reaction. Here we introduced the SLG43 gene of Brassica rapa into a self-incompatible cultivar, Osome, of B. rapa, and examined the effect of this transgene on the SI behavior of the transgenic plants. Preliminary pollination experiments demonstrated that Osome carried S52 and S60, and both were codominant in stigma, but S52 was dominant to S60 in pollen. S43 was found to be recessive to S52 and codominant with S60 in stigma. The nucleotide sequence of SLG43 was more similar to that of SLG52 (87.8% identity) than to that of SLG60 (74.8% identity). Three of the ten primary transformants (designated No. 1 to No. 10) were either completely (No. 9) or partially (No. 6 and No. 7) self-compatible; the SI phenotype of the stigma was changed from S52S60 to S60, but the SI phenotype of the pollen was not altered. In these three plants, the mRNA and protein levels of both SLG43 and SLG52 were reduced, whereas those of SLG60 were not. All the plants in the selfed progeny of No. 9 and No. 6 regained SI and they produced a normal level of SLG52. These results suggest that the alteration of the SI phenotype of the stigma in the transformants Nos. 6, 7, and 9 was the result of specific co-suppression between the SLG43 transgene and the endogenous SLG52 gene. Three of the transformants (Nos. 5, 8 and 10) produced SLG43 protein, but their SI phenotype was not altered. The S60 homozygotes in the selfed progeny of No. 10 which produced the highest level of SLG43 were studied because S43 was codominant with S60 in the stigma. They produced SLG43 at approximately the same level as did S43S60 heterozygotes, but did not show S43 haplotype specificity at the stigma side. We conclude that SLG is necessary for the expression of the S haplotype specificity in the stigma but the introduction of SLG alone is not sufficient for conferring a novel S haplotype specificity to the stigma. PMID- 10480390 TI - Phytochrome types in Picea and Pinus. Expression patterns of PHYA-Related types. AB - Knowledge of the genes in gymnosperms encoding the apoproteins of the plant photoreceptor phytochrome is currently scanty as for gymnosperm nuclear protein coding sequences in general. Here we report two complete cDNA-derived sequences which code for two different types of gymnosperm phytochrome. One sequence stems from Norway spruce (Picea abies) and the other from Scots pine (Pinus sylvestris). More detailed studies have shown that both types of phytochrome gene are present in Norway spruce. From phylogenetic analyses, these types appear to branch off from progenitors that are also the common ancestors of the angiosperm PHYA/PHYC and PHYB/PHYD/PHYE lineages. Partial phytochrome sequences of other gymnosperms cluster with either the one type or the other of the gymnosperm phytochrome genes characterized here. Southern blot analysis of Picea DNA using probes derived from the full-length Picea gene indicated a family of at least five members. Whether they code for new types may be doubted since only two phylogenetic clusters were found. Studies using RNA-PCR of Picea RNA extracted from either light- or dark-grown seedlings indicated that the steady-state levels of the transcripts of two PHYA/C-related genes were hardly affected by light. PMID- 10480391 TI - Altering the 3 UTR endonucleolytic cleavage site of a Chlamydomonas chloroplast mRNA affects 3-end maturation in vitro but not in vivo. AB - The 3' ends of chloroplast mRNAs are produced by the processing of longer precursors. The 3' ends of most plastid mRNAs are located at, or several nucleotides downstream of, stem-loop structures, which act as 3'-end-processing signals and RNA stability elements. In chloroplasts of the green alga Chlamydomonas reinhardtii, 3'-end maturation of atpB mRNA involves endonucleolytic cleavage of the pre-mRNA at an AU-rich site located about 10 nucleotides downstream of the stem-loop structure. This cleavage is followed by exonucleolytic resection to generate the mature 3' end. In order to define critical nucleotides of the endonucleolytic cleavage site, we mutated its sequence. Incubation of synthetic atpB pre-RNAs containing these mutations in a chloroplast protein extract resulted in the accumulation of 3'-end-processed products. However, in two cases where the AU-rich sequence of this site was replaced with a GC-rich one, the 3' end of the stable processing product differed from that of the wild-type product. To examine whether these mutations affected atpB mRNA processing or accumulation in vivo, the endogenous 3' UTR was replaced with mutated sequences by biolistic transformation of Chlamydomonas chloroplasts. Analysis of the resulting strains revealed that the accumulation of atpB mRNA was approximately equal to that of wild-type cells, and that a wild-type atpB 3' end was generated. These results imply that Chlamydomonas atpB 3' processing parallels the situation with other endonucleases such as Escherichia coli RNAse E, where specific sequences are required for correct in vitro processing, but in vivo these mutations can be overcome. PMID- 10480392 TI - Phytoene synthase-2 enzyme activity in tomato does not contribute to carotenoid synthesis in ripening fruit. AB - The characteristic yellow fruit phenotype of the r,r mutant and Psy-1 (phytoene synthase-1) antisense tomatoes is due to a mutated or down-regulated phytoene synthase protein, respectively, resulting in the virtual absence of carotenoids. Based on detailed carotenoid determinations Psy-1 appeared to barely contribute to the formation of carotenoids in chloroplast-containing tissues. Despite the virtual absence of carotenoids in ripe fruit the formation of phytoene in vitro was detected in fruit of both mutants. When [14C]isopentenyl pyrophosphate (IPP) was used as the substrate for phytoene synthase a reduction (e.g. r,r mutant, 5 fold) in the formation of phytoene was observed with an accumulation (e.g. r,r mutant, 2-fold) of the immediate precursor geranylgeranyl pyrophosphate (GGPP). Contrastingly, reduced phytoene synthase activity was not detected when [3H]GGPP was used as the substrate. The profile of phytoene formation during ripening was also different in the down-regulated mutants compared to the wild-type. Using specific primers, RT-PCR analysis detected the presence of Psy-2 transcripts in the down-regulated mutants and wild-type throughout fruit development and ripening. These data were supported by the detection of phytoene synthase protein on western blots. Both GGPP formation and phytoene desaturation were elevated in these mutants. Therefore, it appears that despite the absence of carotenoids in ripe fruit, both the mutants have the enzymic capability to synthesize carotenoids in this tissue. Implications of the data with respect to the regulation of carotenoid formation and the channelling of prenyl lipid precursors in tomato (and its potential manipulation) are discussed. PMID- 10480394 TI - A mini binary vector series for plant transformation. AB - A streamlined mini binary vector was constructed that is less than 1/2 the size of the pBIN19 backbone (3.5 kb). This was accomplished by eliminating over 5 kb of non-T-DNA sequences from the pBIN19 vector. The vector still retains all the essential elements required for a binary vector. These include a RK2 replication origin, the nptIII gene conferring kanamycin resistance in bacteria, both the right and left T-DNA borders, and a multiple cloning site (MCS) in between the T DNA borders to facilitate cloning. Due to the reduced size, more unique restriction sites are available in the MCS, thus allowing more versatile cloning. Since the traF region was not included, it is not possible to mobilize this binary vector into Agrobacterium by triparental mating. This problem can be easily resolved by direct transformation. The mini binary vector has been demonstrated to successfully transform Arabidopsis plants. Based on this mini binary vector, a series of binary vectors were constructed for plant transformation. PMID- 10480393 TI - Interaction between composite elements in the napA promoter: both the B-box ABA responsive complex and the RY/G complex are necessary for seed-specific expression. AB - During seed maturation, the transcriptional activity of napin genes is regulated by developmental signals involving the transcriptional activator ABI3 and abscisic acid (ABA). To localize cis elements involved in the seed-specific activity of the napin napA promoter, a systematic analysis was performed focusing on two major element complexes, the B-box and RY/G. Substitution mutation analysis using promoter-reporter gene fusions in stable transgenic tobacco showed synergistic interactions between elements within these complexes. The distal part of the B-box shows similarities to abscisic acid response elements and the proximal portion contains a CA-rich element. In vitro studies involving Exonuclease III protection and electrophoretic mobility shift assays revealed binding by nuclear proteins to elements within the B-box. The distal and proximal parts of the B-box were found to bind distinct nuclear protein complexes. By gain of-function analysis with a tetramer of the B-box fused to a truncated (-46) cauliflower mosaic virus (CaMV) 35S minimal promoter, it was demonstrated that the B-box mediates strong activity in seeds. Further, it was shown that the elements in the B-box constitute an ABA-responsive complex, since the B-box tetramer mediates ABA-responsiveness in vegetative tissues to a construct containing the CaMV virus 35S enhancer (-343 to -90). Thus, the seed-specific activity of the napA promoter relies on the combinatorial interaction between the RY/G complex and the B-box ABA-responsive complex during the ABA response in seed development. PMID- 10480395 TI - Transcript accumulation and utilization of alternate and non-consensus splice sites in rice granule-bound starch synthase are temperature-sensitive and controlled by a single-nucleotide polymorphism. AB - Granule-bound starch synthase (GBSS), a product of the waxy gene in rice (Oryza sativa L.), is necessary for the synthesis of amylose in the endosperm. In an extended pedigree of 89 rice cultivars, we have previously shown that all cultivars with more than 18% amylose had the sequence AGGTATA at the leader intron 5' splice site, while all cultivars with a lower proportion of amylose had the sequence AGTTATA. This single-nucleotide polymorphism reduces the efficiency of GBSS pre-mRNA processing. It also results in alternate splicing at multiple sites, some of which have non-consensus sequences. Here we demonstrate that this same G-to-T polymorphism is also associated with differential sensitivity to temperature during the period of grain development. Cultivars with the sequence AGTTATA have a substantial increase in accumulation of mature GBSS transcripts at 18 degrees C compared to 25 or 32 degrees C. The selection of leader intron 5' splice sites is also affected by temperature in these cultivars. A 5' splice site -93 upstream from that used in high-amylose varieties predominates at 18 degrees C. At higher temperatures there is increased utilization of a 5' splice site at I and a non-consensus site at +1. Potential implications of differential 5' splice site selection and associated differences in 3' splice site selection on transcript stability and translational efficiency are discussed. PMID- 10480396 TI - Salt-specific regulation of the cytosolic O-acetylserine(thiol)lyase gene from Arabidopsis thaliana is dependent on abscisic acid. AB - The expression of Atcys-3A gene coding for cytosolic O-acetylserine(thiol)lyase, a key enzyme in cysteine biosynthesis, from Arabidopsis thaliana is significantly induced by exposure to salt and heavy-metal stresses. Addition of NaCl to mature plants induced a rapid accumulation of the mRNA throughout the leaf lamina and roots, and later on in stems, being mainly restricted to vascular tissues. The salt-specific regulation of Atcys-3A was also mediated by abscisic acid (ABA) since: (1) exogenous addition of ABA to the culture medium mimicked the salt induced plant response by raising the level of Atcys-3A transcript, and (2) Arabidopsis mutants aba-1 and abi2-1 were not able to respond to NaCl. Our results suggest that a high rate of cysteine biosynthesis is required in Arabidopsis under salt stress necessary for a plant protection or adaptation mechanism. This hypothesis was supported by the observation that intracellular levels of cysteine and glutathione increased up to 3-fold after salt treatment. PMID- 10480398 TI - Mixed chimerism as an approach for the induction of transplantation tolerance. PMID- 10480397 TI - Characterisation of the PsbX protein from Photosystem II and light regulation of its gene expression in higher plants. AB - The location and expression of the previously uncharacterised photosystem II subunit PsbX have been analysed in higher plants. We show that this protein is a component of photosystem II (PSII) core particles but absent from light harvesting complexes or PSII reaction centres. PsbX is, however, localised to the near vicinity of the reaction centre because it can be cross-linked to cytochrome b559, which is known to be associated with the D1/D2 dimer. We also show that the expression of this protein is tightly regulated by light, since neither protein nor mRNA is found in dark-grown plants. PMID- 10480399 TI - TGF-beta quantitation can be tricky. PMID- 10480400 TI - Understanding "marginal" liver grafts. PMID- 10480401 TI - Technical-variant liver grafts in pediatric liver transplantation: back to the future. PMID- 10480402 TI - Allele-specific and peptide-dependent recognition of swine leukocyte antigen class I by human cytotoxic T-cell clones. AB - BACKGROUND: The T-cell mediated immune responses play a major role in xenograft rejection. However, the mechanisms behind human T-cell recognition of porcine xenoantigens remain to be elucidated. METHODS: Human CD8+ T-cell lines were generated against porcine aortic endothelial cells (PAECs) from y/y and z/z haplotypes of Yucatan inbred swine. T-cell clones were obtained by limiting dilution. The human T-cell receptor (TCR)-swine leukocyte antigen (SLA) class I interaction was characterized. RESULTS: The human CD8+ T-cell mediated direct recognition of PAECs was SLA haplotype-specific. The haplotype specificity was restricted by the SLA class I allelic polymorphism. To characterize the role of SLA-bound peptides in the human TCR-SLA class I interaction, we stripped peptides from SLA molecules by a brief acid treatment. Using z/z-specific CD8+ T cells as effectors, we demonstrated that the acid-treatment, which stripped SLA molecules of bound peptides, decreased the lysis of PAECs by 72%. Addition of peptides eluted from affinity purified z/z SLA class I molecules, but not from the irrelevant y/y SLA class I, restored the lysis of acid-treated z/z PAECs. In addition, the lysis of a human HLA class I negative cell line, 721.221, transfected with a relevant SLA class I allele derived from the z/z haplotype, was significantly increased with the addition of relevant z/z peptides. These experiments indicated that both SLA class I and bound peptides were required for recognition by human CD8+ T cells. Cloning studies identified two groups of xenoreactive T-cell clones. Group I clones recognized distinct porcine peptides in the context of SLA class I molecules, whereas group II clones recognized human endogenous cross-reactive peptides presented by SLA class I. CONCLUSIONS: Our results demonstrated that, despite the differences in MHC molecules between species, human T-cell recognition of porcine MHC is similar to direct allo recognition, that is, human TCR recognizes xenogeneic SLA-peptide complexes. PMID- 10480403 TI - Combined histocompatibility leukocyte antigen-matched donor bone marrow and renal transplantation for multiple myeloma with end stage renal disease: the induction of allograft tolerance through mixed lymphohematopoietic chimerism. AB - BACKGROUND: Experimental and clinical evidence has demonstrated that the establishment of allogeneic chimerism after bone marrow transplantation may provide donor-specific tolerance for solid organ allografts. METHODS: Based on the preliminary results of a clinical trial using nonmyeloablative preparative therapy for the induction of mixed lymphohematopoietic chimerism, we treated a 55 year-old woman with end stage renal disease secondary to multiple myeloma with a combined histocompatibility leukocyte antigen-matched bone marrow and renal transplant after conditioning with cyclophosphamide, antithymocyte globulin, and thymic irradiation. RESULTS: The posttransplant course was notable for early normalization of renal function, the absence of acute graft-versus-host disease, and the establishment of mixed lymphohematopoietic chimerism. Cyclosporine, which was the only posttransplant immunosuppressive therapy, was tapered and discontinued on day +73 posttransplant. No rejection episodes occurred, and renal function remains normal on day + 170 posttransplant (14 weeks after discontinuing cyclosporine). Although there is presently no evidence of donor hematopoiesis, there is evidence of an ongoing antitumor response with a recent staging evaluation showing no measurable urine kappa light chains. The patient remains clinically well and is off all immunosuppressive therapy. CONCLUSION: This is the first report of the deliberate induction of mixed lymphohematopoietic chimerism after a nonmyeloablative preparative regimen to treat a hematological malignancy and to provide allotolerance for a solid organ transplant. PMID- 10480404 TI - The effect of thymectomy on tolerance induction and cardiac allograft vasculopathy in a miniature swine heart/kidney transplantation model. AB - BACKGROUND: We have previously demonstrated that MHC class I disparate hearts transplanted into miniature swine treated with a short course of cyclosporine developed florid cardiac allograft vasculopathy (CAV) and were rejected within 55 days. However, when a donor-specific kidney is cotransplanted with the heart allograft, recipients become tolerant to donor antigen and accept both allografts long-term without the development of CAV. In the present study, we have investigated the role of the host thymus in the induction of tolerance and prevention of CAV in heart/kidney recipients. METHODS: Total thymectomies were performed in six animals (postoperative day [POD]-21), which on day 0 received either an isolated MHC class I disparate heart allograft (n=3) or combined class I disparate heart and kidney allografts (n=3), followed in both cases by a 12-day course of cyclosporine (POD 0-11). Graft survival and the development of CAV in these thymectomized recipients were compared to the same parameters in non thymectomized, cyclosporine-treated recipients bearing either class I disparate heart allografts (n=5) or heart and kidney allografts (n=4). RESULTS: In the group of animals bearing isolated class I disparate heart allografts, the thymectomized recipients rejected their allografts earlier (POD 8, 22, 27) than the non-thymectomized recipients (POD 33,35,45,47,55). The donor hearts in both the thymectomized and non-thymectomized animals developed florid CAV. In the group of animals bearing combined class I disparate heart and kidney allografts, the nonthymectomized recipients accepted both donor organs long term with no evidence of CAV. In contrast, none of the thymectomized heart/kidney recipients survived >100 days, and they all developed the intimal proliferation of CAV. CONCLUSION: Thymic-dependent mechanisms are necessary for the induction of acquired tolerance and prevention of CAV in porcine heart/kidney recipients. PMID- 10480405 TI - The influence of combined trophic factors on the success of fetal pancreas grafts. AB - BACKGROUND: Fetal pancreas (FP) has the capacity for abundant proliferation and beta cell differentiation. Insulin-like growth factor-1 (IGF-1) promotes FP engraftment in the i.m. site and reversal of diabetes in a rodent model. However, reversal of diabetes by an FP transplant in rats under the influence of IGF-1 is still an inefficient process requiring multiple FP grafts and a prolonged latent period. Numerous other growth and differentiation factors, which include platelet derived growth factor (PDGF), vascular endothelial growth factor, endothelial cell growth factor-alpha and pancreatic islet neogenesis-associated protein, have been implicated in beta cell neogenesis and proliferation. We have analyzed the in vivo role of these growth factors in FP engraftment and reversal of streptozotocin-induced diabetes in rats. METHODS: IGF-1 alone or in combination with other trophic factors was locally administered to eight FP isografts in the thigh muscle of diabetic rats. RESULTS: Diabetes was reversed in a mean of 60+/ 26 days in 11 of 11 animals treated with IGF-1. PDGF alone did not promote reversal of diabetes; however, PDGF + IGF-1 resulted in euglycemia in 6 of 6, with a mean of 36+/-14 days (P<0.05). Islet neogenesis-associated protein +IGF-1 resulted in reversal of diabetes in 6 of 6 rats with a mean interval of 50+/-10 days. Vascular endothelial growth factor or endothelial cell growth factor-alpha + IGF-1 provided no advantage compared with IGF-1 alone. CONCLUSIONS: These results demonstrate that IGF-1 is a potent trophic factor for transplanted FP and that PDGF acts synergistically with IGF-1 to promote reversal of diabetes by transplanting FP. PMID- 10480406 TI - Laparoscopic versus open donor nephrectomy: comparing ureteral complications in the recipients and improving the laparoscopic technique. AB - BACKGROUND: Laparoscopic live donor nephrectomy (LDN) is a recently developed procedure, the performance of which needs to be studied. Given the reported advantages in the donors, this study looks at graft outcome and ureteral complications in recipients of kidneys procured by open donor nephrectomy (ODN) versus LDN. METHODS: The LDN recipients consisted of 193 patients since 3/27/96. A total of 168 ODN recipients from 1991 to 1998 served as controls. Immunosuppression protocols were similar for both groups. RESULTS: Two-year graft survival for LDN and ODN was 98% and 96%, respectively. Two-year patient survival for LDN and ODN was 98% and 97%, respectively. The incidence of delayed graft function and mean serum creatinine at 3 and 12 months was similar in both groups. However, the number of ureteral complications that required operative repair was significantly higher for LDN recipients compared to ODN recipients, 7.7% (n=15) vs. 0.6% (n=1) respectively (P=0.03). Ureteral stenting was required in an additional 3.1% (n=6) of LDN and 2.4% (n=4) of ODN (P=NS). There was, however, a learning curve with time. For the first 130 LDN patients, a total of 20 ureteral complications were recorded, whereas only one occurred in the more recent 63 patients (P=0.03). CONCLUSIONS: The higher ureteral complication rate in LDN recipients has improved over time as technical causes have been identified. We have noted significant improvement in ureteral viability by using the endogastrointestinal anastomosis instrument on the ureter and peri-ureteral tissue. LDN is therefore an excellent alternative to ODN. Identification of hazards unique to this technique is critical before its broader application. PMID- 10480407 TI - The clinical importance of choosing the right assay for detection of HLA-specific donor-reactive antibodies. AB - BACKGROUND: We found earlier that there is a close clinical correlation between the presence of histocompatibility leukocyte antigens (HLA) class I-specific donor-reactive antibodies in cross-match serum and a significantly higher frequency of early acute rejection episodes and graft loss during the first year after the transplant. METHODS: Specificity determinations of donor-reactive antibodies present in the cross-match serum before allogeneic kidney transplants were performed. In the present study, we compared the suitability and efficiency of (a) platelet absorptions, (b) blocking with anti-HLA monoclonal antibodies in the microcytotoxicity assay, and (c) donor-specific HLA antigen-coated magnetic microbeads in flow cytometric assays for the definition of clinically relevant HLA antibodies and their correlation with early acute rejections and early graft loss. RESULTS: We found that the microlymphocytoxicity test using donor splenic B lymphocytes often gave positive reactions in the absence of class I or class II antibodies; in other words, a high frequency of false positive reactions was observed. Flow cytometric tests are more sensitive than microlymphocytotoxicity, not only because they are more sensitive, but also because noncomplement-binding antibodies are detected. Platelet absorptions, which detect only reactivity against HLA class I antigens, is insufficient for use in specificity determinations of donor-reactive antibodies. We found that a positive test for HLA antibodies using paramagnetic beads coated with solubilized donor-derived HLA antigens (class I or class II) correlates with early immunological complications after a transplant (P<0.001). CONCLUSION: Assays to determine the specificity of donor-reactive antibodies are now available for use during an acute transplant situation. The introduction of such methods is expected to enhance graft survival and to reduce significantly the frequencies of acute rejections episodes after a transplant. PMID- 10480408 TI - Attitudes of patients before and after transplantation towards various allografts. AB - BACKGROUND: The presence of an allogeneic graft inside the body may have psychological impact on transplant patients. It was the aim of this study to evaluate the attitude of patients before and after different types of organ transplantation towards organ allografts. METHODS: A total of 1,049 patients (722 patients after and 327 patients on the waiting list for either kidney, liver, heart, or lung transplantation) under care of a single transplant center were studied using a questionnaire with blinded analysis of the data. Mental condition of the patients, their attitude towards the allograft and its donor, emotional stress caused by a graft, and correlation of the attitudes with clinical and demographic parameters were analyzed. RESULTS: The self-reported mental condition of the patients was markedly and consistently better after organ transplantation; 27% of patients before and 60% after transplantation were in good mental condition. Sixty-two percent of transplant patients considered the graft as their own organ, 37% regarded it as a foreign organ now belonging to their body, and 1% considered it as a foreign body; among waiting list patients, 40%, 55%, and 5% assumed they would perceive their graft accordingly. The graft caused considerable emotional stress for 2% of transplant patients, whereas for 70% it did not cause any stress; the latter was assumed by 47% of patients before transplantation. Eleven percent of transplant patients frequently think about the origin of their graft, and 30% would like to have information about their donor. Knowledge about different religion, opposite sex, homosexuality, suicidal death, and age above 65 years of their donor would be of moderate or major concern for 0%, 3%, 21%, 24%, and 38% of the patients, respectively. CONCLUSIONS: The comprehensive survey shows that transplant patients incorporate their graft well into their body image. Emotional stress caused by the graft is very low and is generally less than assumed before transplantation. Knowledge about certain characteristics of the donor may cause increased concerns in some patients. PMID- 10480409 TI - The influence of histocompatibility on graft rejection and graft survival within a single center population of heart transplant recipients. AB - BACKGROUND: We report a consecutive single center series of 261 patients who received first orthotopic heart transplants from 1986 to 1997. The 1- and 5-year graft survivals were 78 and 68%. The influence of histocompatibility was investigated by comparing graft survival and numbers of treated rejection episodes with HLA-A, -B, and -DR mismatches over different time periods. FINDINGS: Recipients with six mismatches for HLA-A+-B+-DR combined (13.4%) had reduced survival at 7 years (47%) when compared with other recipients (64%). In the first year of transplant, recipients with four HLA-A+-B mismatches had significantly reduced actuarial graft survival (P=0.03) with the greatest influence apparent at 6 months [0-3 mismatches (n=193) 85% versus 4 mismatches (n=68) 69%; P=0.005, OR=2.1]. For 182 recipients with functioning hearts at 1 year, the number of rejection episodes treated within this time was strongly influenced by HLA-DR mismatch [0 DR mismatch (n=15) mean 1.2 rejection episodes versus 1 DR mismatch (n=76) mean 2.7 rejection episodes versus 2 DR mismatches (n=91) mean 3.8 rejection episodes: P=0.0002]. Of these 182 transplants, recipients who had more than four treated rejection episodes during the first year had a significantly reduced 7- year survival [<5 rejection episodes (n=133) 85% versus more than four rejection episodes (n=49) 66%; P=0.02, OR=3.4], as did those with two HLA-DR mismatches [0+1 mismatch (n=91) 87% versus 2 mismatches (n=91) 70%; P<0.05, OR=2.4]. INTERPRETATION: We show that graft loss in the first 6 months of transplant is significantly influenced by four HLA-A+-B mismatches. HLA-DR mismatch significantly increases the number of rejection episodes within the first year, without influencing graft survival. After 12 months both >4 rejection episodes in the first year and two HLA-DR mismatches are markers for late graft loss. We postulate that immunological graft loss in the first 6 months is dominated by the direct allorecognition pathway driven by HLA-DR mismatch. This mechanism is later lost or suppressed. Our data highlight HLA-DR mismatch as a marker for late graft loss and we show an advantage to avoiding transplanting hearts with six HLA-A+-B+-DR mismatches and to minimizing HLA-DR mismatches whenever possible. PMID- 10480410 TI - Can hepatitis B core antibody positive livers be used safely for transplantation: hepatitis B virus detection in the liver of individuals who are hepatitis B core antibody positive. AB - A major impediment to the wider application of clinical liver transplantation is the paucity of acceptable organs. Most centers refuse organs that come from donors who are hepatitis B core antibody positive because of a fear of transmission of hepatitis B virus (HBV) infection to the recipient. The risk related to the use of such donor organs has never been assessed in an ordered manner. The presence or absence of polymerase chain reaction detectable HBV-DNA in liver tissue of individuals undergoing liver biopsy for clinical reasons was assessed in 133 consecutive patients. A total of 8.2% of these livers resulted positive for HBV-DNA; interestingly the rate was higher among those who were hepatitis B surface antibody positive (12.5%) as compared to those without detectable hepatitis B surface antibody (5.7%). These data provide measures of putative risk for HBV infection in liver transplant recipients associated with the use of organs obtained from a hepatitis B core antibody positive donor. PMID- 10480411 TI - Prevention of osteoporosis after cardiac transplantation: a prospective, longitudinal, randomized, double-blind trial with calcitriol. AB - BACKGROUND: Accelerated bone loss is a well-recognized complication after cardiac transplantation (HTx) due to immunosuppressive therapy. The purpose of this prospective, longitudinal, randomized, placebo-controlled, double-blind study was to investigate the effect of calcitriol (1,25-dihydroxyvitamin D3) in the prevention of bone loss and fracture rate after HTx. METHODS: Basic therapy included 1000 mg of calcium daily and sex hormone replacement in hypogonadal patients. A total of 132 patients (111 male, 21 female; mean age: 51+/-10 years; 35+/-25 months after HTx) were randomized to 0.25 microg of calcitriol or placebo. Bone mineral density (BMD, g/cm2; T score, %) of the lumbar spine and x rays for the assessment of vertebral fractures were performed at baseline and after 12, 24, and 36 months. Biochemical indexes of mineral metabolism were measured every 3 months. RESULTS: Overall BMD was significantly decreased after HTx (T score 87+/-13%). BMD increased continuously within the study period in the calcitriol group (1 year: 2.2+/-4.8%; 2 years: 3.9+/-5.4%; 3 years: 5.7+/-4.4%) as well as in the placebo group (1 year: 1.8+/-4.9%; 2 years: 3.7+/-6.5%; 3 years: 6.1+/-7.8%) without statistical difference between the groups. Fracture incidence was low during the study interval (1 year: 2.0%; 2 years: 3.4%; 3 years: 0%). Hypogonadism (20%) was associated with a lower BMD (78+/-12% vs. 88+/ 12%; P<0.01) and a higher increase (35%) after hormone replacement in comparison to normogonadal patients. Increased intact parathyroid hormone and bone resorption markers decreased significantly during therapy. CONCLUSIONS: Calcium supplementation and sex hormone replacement in hypogonadism proved a sufficient long-term prevention therapy to improve decreased BMD and to prevent fractures after HTx. Besides immunosuppression, both concomitant hypogonadism and secondary hyperparathyroidism play a major role in the long-term bone loss and should therefore be monitored and treated adequately. Low-dose calcitriol demonstrated no significant extra benefit regarding BMD and fracture rate in the long-term period after HTx. PMID- 10480412 TI - Poor outcome in patients with diabetes mellitus undergoing liver transplantation. AB - BACKGROUND: Relatively few studies have examined the influence of pretransplant diabetes on survival after an orthotopic liver transplant (OLT), and those published to date show only minor increases in infection rates among diabetics and no increase in mortality. METHODS: We examined the effect of diabetes mellitus on survival after OLT. 1005 adults underwent OLT between 1982 and May 1997. Seventy-eight patients with pretransplant diabetes mellitus (7.8% of all OLT, 38 insulin treated, 25 tablet treated, 15 diet controlled) were identified and compared with controls matched for age, sex, and date of first transplant and also with all nondiabetic adult liver recipients undergoing OLT during the same period. RESULTS: In patients undergoing OLT survival was worse in diabetics than in the comparison group (P=0.002) and vs. all adult nondiabetics undergoing (n=927) (P=0.004); in diabetics with alcoholic liver disease (ALD) vs. all nondiabetics with alcoholic liver disease (P= <0.0001); and in insulin-treated compared with non-insulin-treated diabetics (P=0.05). Multivariate analysis showed type of diabetes (P=0.001) and ALD (P=0.024) to be the most significant independent variables adversely affecting survival. Survival in diabetics undergoing OLT could be further stratified according to whether diabetics were insulin treated. CONCLUSIONS: Poorer outcome in the diabetics undergoing OLT, particularly in those with ALD, suggests the need for a more detailed pre-OLT assessment of these patients, particularly those with insulin and tablet controlled diabetes. PMID- 10480413 TI - Increased incidence of postoperative infections associated with peritoneal dialysis in renal transplant recipients. AB - BACKGROUND: Infection is a frequent postoperative complication in renal transplant recipients. However, little information is available concerning the effect of pretransplantation dialysis modality on posttransplantation complications including infection. We therefore evaluated the effect of hemodialysis (HD) versus peritoneal dialysis (PD) on the incidence of postoperative infection as well as several other posttransplantation outcomes. METHODS: A retrospective analysis was performed using medical records covering the period 30 days after transplantation of 156 dialysis patients who underwent renal transplantation at a single center during a 22-month period. Of these patients, 103 received only HD, 32 received only PD, 13 received PD in the past and HD immediately before transplantation (PH/HD), and 8 received HD in the past and PD immediately before transplantation (HD/PD). The presence of culture-proven infection, types of infecting organisms, length of initial hospital stay, and incidence of rejection during the first 30 days after transplantation were determined for each patient. RESULTS: All groups were similar with regard to age, race, gender, underlying disease, donor type, incidence of delayed graft function, and perioperative antibiotic prophylaxis. There were more infectious complications within 30 days after transplantation in patients on PD just prior to transplantation (PD and HD/PD) than in HD patients (67.5% vs. 25.9%, P<0.00001). When types of infectious organisms were assessed, PD patients were found to have a greater incidence of infections with microorganisms that colonize human skin (P<0.0001). The median length of hospital stay was 3 days longer for PD patients and 6.5 days longer for HD/PD patients than for patients receiving HD (P=0.01 and 0.04), and PD and HD/PD patients were more likely to have an episode of rejection than HD patients (P=0.02). CONCLUSIONS: Renal replacement therapy with PD immediately before transplantation negatively affects outcome as compared with HD, predisposing patients to a greater incidence of postoperative infections and rejection and a longer hospital stay. Further study in a randomized controlled trial may help determine how adjustment of the dialysis method can optimize transplantation outcome. PMID- 10480414 TI - Analysis of survival and morbidity after pediatric liver transplantation with full-size and technical-variant grafts. AB - BACKGROUND: To alleviate the shortage of size-matched whole-donor organs, too large-for-size cadaveric donor grafts are modified by liver resection techniques. These modifications result in technical-variant liver transplantation (TVLTx). Patient and graft survival rates after TVLTx are considered comparable to those after full-size liver transplantation (FSLTx). However, morbidity after TVLTx is often underexposed. The aim of this study was to analyze the results of FSLTx and TVLTx in terms of patient and graft survival rates and morbidity. METHODS: A consecutive series of 97 primary and elective pediatric liver transplantations performed in a single center was retrospectively analyzed. Forty-seven children had a FSLTx and 50 a TVLTx (38 reduced-size liver grafts and 12 split-liver grafts). The overall median follow-up period was 3.5 years. RESULTS: There were no differences in patient and graft survival rates between FSLTx and TVLTx. However, after TVLTx there was a significantly higher complication rate (1.42 vs. 0.81 after FSLTx). TVLTx is more hampered by biliary complications (30% vs. 17%), expressed by a higher incidence of cholangitis and leakage of bile. These complications led to a significantly higher incidence of sepsis (44% vs. 19%) and a significantly higher intervention rate (0.40 vs. 1.28) after TVLTx. There was no difference in the incidence of retransplantations between FSLTx and TVLTx. CONCLUSIONS: Both FSLTx and TVLTx offer the same prognosis in terms of patient and graft survival rates for children after a primary and elective liver transplantation. However, TVLTx has a higher morbidity. PMID- 10480415 TI - A humanized anti-CD3 antibody, HuM291, with low mitogenic activity, mediates complete and reversible T-cell depletion in chimpanzees. AB - BACKGROUND: An anti-CD3 antibody that reduces cytokine release syndrome (CRS) while maintaining immunosuppression would be a major advance in the treatment of acute allograft rejection. A humanized (Hu) anti-CD3 IgG2 Ab, HuM291 gamma2 M3 (HuM291; Protein Design Labs, Inc., Mountain View, CA), was engineered with mutations in the upper CH2 region of the Fc domain. The mutations were intended to reduce affinity for Fcgamma receptors, thought to be relevant to CRS. METHODS: In vitro studies using chimpanzee peripheral blood mononuclear cells (PBMCs) were conducted to characterize HuM291 and to establish an animal model. A multidose study was conducted in chimpanzees to evaluate the safety, pharmacokinetics, immunomodulatory activity, and immunogenicity of HuM291, when administered at doses ranging from 0.1 to 10 mg. RESULTS: HuM291 bound to and effectively downmodulated CD3 from chimpanzee PBMCs and stimulated substantially less cytokine secretion and proliferation of chimpanzee PBMCs compared with OKT3 (Orthoclone OKT3; Ortho Pharmaceutical Corp., Raritan, NJ). Multiple doses of HuM291 (0.1, 1.0, or 10 mg/dose) were not associated with adverse events, signs of toxicity, or CRS, despite cytokine release. HuM291 exhibited a long elimination t1/2 (81.5 hr) and, after three 10-mg doses, sustained serum concentrations > 1000 ng/ml were maintained for 1 week. Multiple 10-mg doses induced complete depletion of circulating CD2+CD3+ T cells for up to 10 days after the last dose; T cells recovered by Day 28. Anti-HuM291 Abs were observed in only 4 of 12 animals and were transient in 2 of those animals. CONCLUSIONS: In vitro, HuM291 is substantially less mitogenic than OKT3. In chimpanzees, HuM291 effectively depleted peripheral T cells without eliciting clinical signs of CRS, and recovered T cells were functionally normal. PMID- 10480416 TI - Patterns of immune responses evoked by allogeneic hepatocytes: evidence for independent co-dominant roles for CD4+ and CD8+ T-cell responses in acute rejection. AB - INTRODUCTION: This is the first in a series of reports that characterizes immune responses evoked by allogeneic hepatocytes using a functional model of hepatocyte transplantation in mice. METHODS: "Donor" hepatocytes expressing the transgene human alpha-1-antitrypsin (hA1AT-FVB/N, H2q) were transplanted into C57BL/6 (H2b) or MHC II knockout (H2b) hosts treated with anti-CD4, anti-CD8, or a combination of anti-CD4 and anti-CD8 monoclonal antibodies (mAbs). Hepatocyte rejection was determined as a loss of circulating ELISA-detectable transgene product (hA1AT). In addition, some C57BL/6 mice underwent transplantation with FVB/N heterotopic cardiac allografts and were treated with anti-CD4 mAb. Cardiac allograft rejection was determined by palpation. Graft recipients were tested for donor reactive alloantibodies and donor-reactive delayed-type hypersensitivity (DTH) responses. RESULTS: The median survival time (MST) of allogeneic hepatocytes in normal C57BL/6 mice was 10 days (no treatment), 10 days (anti-CD4 mAb), 14 days (anti-CD8 mAb), and 35 days (anti-CD4 and anti-CD8 mAbs). The MST of hepatocytes in B6 MHC class II knockout mice was 10 days (no treatment) and 21 days (anti-CD8 mAb). The MST of cardiac allografts was 11 days (no treatment) and >100 days (anti-CD4 mAb). Donor-reactive DTH responses were readily detected in both untreated and mAb-treated recipients. Donor-reactive alloantibody was barely detectable in untreated hosts. CONCLUSIONS: These studies demonstrate that allogeneic hepatocytes are highly immunogenic and stimulate strong cell-mediated immune responses by both CD4+ and CD8+ T cells, even when treated with agents that can cause acceptance of cardiac allografts. Indeed, CD4+ or CD8+ T cells seem to independently cause hepatocellular allograft rejection. Allogeneic hepatocytes evoked strong donor-reactive DTH responses but were poor stimuli for donor-reactive antibody production. This is an unusual pattern of immune reactivity in allograft recipients. PMID- 10480418 TI - Use of octogenarian livers safely expands the donor pool. AB - BACKGROUND: The increasing number of recipients on the waiting list for orthotopic liver transplantation (OLT) and the scarcity of donors contribute to recipient pretransplantation mortality. One important measure to increase the donor liver pool would be to accept the previously discarded donors who are more than 80 years old. METHODS: From November 1996 to May 1998, four liver grafts from octogenarian donors (89, 87, 82, and 85 years old, respectively) were used for OLT. Pretransplantation donor and recipient characteristics and the evolution of recipients after OLT were analyzed. RESULTS: The donors did not present cardiac arrest or hypotension, and only low doses of vasopressors were required in three of them. Intensive care unit stay of the donors was from 12 to 24 hr. Cold ischemia time was from 4 hr to 8 hr 40 min. Mild microsteatosis was present in three donors and associated macrosteatosis of < 10% in one of these. Macroscopic appearance and consistency were normal in all four grafts. Posttransplantation evolution and follow-up were uneventful. Three recipients were alive and well at 24, 16, and 7 months; the second of these died at 16 months of recurrent viral C cirrhosis after a first OLT. CONCLUSIONS: The liver donor pool can be increased if liver grafts are accepted without an age limit but in good condition (hemodynamic stability, short intensive care unit stay, good liver function, soft consistency, cold ischemia time <9 hr, and no severe steatosis). Octogenarian donors should be individually assessed in the absence of these ideal conditions. PMID- 10480417 TI - HuM291, a humanized anti-CD3 antibody, is immunosuppressive to T cells while exhibiting reduced mitogenicity in vitro. AB - BACKGROUND: OKT3, a mouse monoclonal antibody (Ab) specific for the human CD3 complex on T cells, is a potent immunosuppressive agent used for the treatment of acute allograft rejection. The utility of the drug has been limited by a neutralizing anti-mouse Ab response and adverse side effects resulting from T cell activation and systemic cytokine release. T cell activation is caused by OKT3-mediated cross-linking of T cells and Fc receptor-bearing cells. Studies in the mouse model have shown that global T cell activation is not necessary for immunosuppression, as Fc receptor-nonbinding anti-CD3 Abs can suppress graft rejection in the absence of the activation effects seen with Fc receptor-binding Abs. Thus, a humanized anti-CD3 antibody with a low affinity for Fc receptors might improve immunosuppressive therapy by reducing the side effects associated with OKT3. METHODS: We developed a mouse monoclonal Ab, M291, which competes with OKT3 for binding to T cells. Humanized, complementary-determining region-grafted versions of M291 featuring various Fc were engineered, including a previously described IgG2 mutant deficient in Fc receptor binding (HuM291). RESULTS: Compared with OKT3 and HuM291-IgG1, HuM291 was significantly less mitogenic to T cells in vitro and induced the release of much lower levels of the cytokines tumor necrosis factor-alpha, interferon-gamma, and interleukin-10. Despite this reduction in T cell activation, HuM291 retained the ability to modulate the CD3 complex and inhibit the mixed lymphocyte reaction. CONCLUSIONS: When evaluated in vivo, HuM291 may be an immunosuppressive agent associated with less of the acute toxicity and immunogenicity seen with OKT3 therapy. PMID- 10480420 TI - Clear benefit of mycophenolate mofetil-based triple therapy in reducing the incidence of acute rejection after living donor renal transplantations. AB - BACKGROUND: According to a pooled analysis of three randomized clinical studies concerning the prevention of acute rejection in cadaveric renal transplantation, mycophenolate mofetil (MMF) proved superior to azathioprine or placebo in conjunction with cyclosporine (CsA) and steroids. MMF-treated patients showed reduced incidence and severity of acute rejection, similar graft survival, and better graft function over 12 months. However, the multicenter trials did not include the Asian recipients of living donor kidneys. METHODS: To assess the efficacy of MMF as the third component of a triple therapy in addition to CsA Neoral and steroids in living donor renal transplantation recipients in Asians, a total of 100 recipients were randomized to receive CsA-Neoral and steroids (control group, n=50), or MMF-based triple therapy (1.0 g of MMF twice daily from postoperative day 2, MMF group, n=50). The dosing plan for Neoral and steroids was essentially same between groups. During 12 months of follow-up, we compared the incidence of acute rejection, adverse events such as infections, and 12-month actual graft and patient survival. RESULTS: The graft and patient survival at 1 year was excellent in both groups: 96/98% in the control group and 98/100% in the MMF group, respectively. MMF significantly reduced the proportion of patients with at least one episode of acute rejection (34% in the control group vs. 14% in the MMF group), cumulative incidence of acute rejection episodes (46% vs. 16%), and requirement of antilymphocyte antibody (21.7% vs. 12.5%). In the MMF group, viral infection such as herpes zoster or chicken pox was more prevalent than in the control group. CONCLUSIONS: Like cadaveric renal transplantation, this open clinical trial showed MMF to be effective in reducing the incidence and severity of acute rejection if used in conjunction with Neoral and steroids after living donor renal transplantation in Asian ethnicity. PMID- 10480419 TI - Heterozygous prothrombin gene mutation: a new risk factor for early renal allograft thrombosis. AB - BACKGROUND: Underlying thrombophilic disorders increase the risk of early allograft loss after renal transplantation. We report three cases of early graft thrombosis in two carriers of a recently discovered prothrombotic variation of the prothrombin gene. CASE REPORTS: The first patient, an adolescent girl, developed multiple thrombotic shunt occlusions after the initiation of hemodialysis until continuous cumarin anticoagulation was instituted. During living-related kidney transplantation, peracute thrombosis of the renal arteries and veins occurred during surgery despite excellent intraoperative conditions and continuous low-dose heparinization. A few hours after reperfusion of the organ by immediate thrombectomy and intrarenal fibrinolysis, an irreversible rethrombosis occurred. A detailed evaluation of the coagulation system showed highly elevated prothrombin protein activity and concentrations. A heterozygous G-->A transition at position 20210 of the prothrombin gene was identified. Hemodialysis was resumed using recombinant hirudin, a direct and selective thrombin inhibitor, as an anticoagulant. The second patient, a girl with end-stage renal failure due to atypical hemolytic uremic syndrome, lost two cadaver kidney allografts, each time by massive thrombosis a few days after transplantation. In this patient also, elevated prothrombin activity and concentrations were present and a heterozygous G-->A transition at position 2210 of the prothrombin gene was detected. CONCLUSIONS: The prothrombin gene mutation is a new risk factor for thrombotic complications both on hemodialysis and after renal transplantation. It may be useful to screen for this disorder in the pretransplant thrombophilia work-up. PMID- 10480421 TI - Laparoscopic living donor nephrectomy: advantages of the hand-assisted method. AB - INTRODUCTION: The laparoscopic technique for living donor nephrectomy is a technically difficult procedure that has not yet gained widespread acceptance in the transplant community. The procedure may be more acceptable if alterations to the technique made it easier to perform and decreased operative times. METHODS: In August 1998, we altered the laparoscopic procedure to include the use of a device allowing hand assistance. Subsequently, all living donor nephrectomies have been done using the hand-assisted method. In this article, the results of 10 cases performed using the original laparoscopic technique are compared with the results of 12 cases using the hand-assisted technique, and a brief description of modifications to the original technique is given. RESULTS: No patients where turned down as living donors, and no contraindications to the pure or hand assisted laparoscopic techniques where found. The hand-assisted technique significantly reduced the operative time (2.02+/-0.44 vs. 3.12+/-0.36 hr, P<0.05) and the warm ischemic time (1.23+/-0.54 vs. 3.91+/-0.53 min, P<0.05). The length of stay and recovery time to normal activities were not different between the pure laparoscopic and hand-assisted groups. CONCLUSION: The advantages of the hand-assisted technique include the ability to use tactile sense to facilitate dissection, retraction, and exposure. In addition, the final stages of vascular stapling and kidney removal are more sure and rapid. The modifications of the laparoscopic technique presented here provide measurable and subjective improvements to laparoscopic living donor nephrectomy. The hand-assisted method of laparoscopic nephrectomy may make the operation available to more transplant centers. PMID- 10480422 TI - Blood levels of TGFbeta1 in liver transplant recipients receiving either tacrolimus or micro-emulsified cyclosporine. AB - BACKGROUND: Transforming growth factor beta-1 (TGFbeta1) is pro-fibrotic in addition to being a potent immunosuppressive cytokine. Cyclosporine (cyclosporin A[CsA]) has been found to increase circulating TGFbeta1 levels in patients (1, 2). To determine whether tacrolimus (FK506) similarly increases TGFbeta1 we have measured TGFbeta levels in blood samples from liver graft recipients who were of known TGFbeta1-responder status. METHODS: Sequential serum and plasma samples were obtained from liver transplant recipients in the UK trial of tacrolimus versus microemulsified CsA, with a follow up period of between 50 and 265 days. Twelve patients received CsA and 13 received tacrolimus. Active and total TGFbeta1 protein were measured and plasma beta thromboglobulin (betaTG) levels were used as an indirect indication of platelet-derived TGFbeta contamination of samples. RESULTS: We found no correlation between trough drug levels and active TGFbeta1 levels in serum of either set of patients. Plasma beta thromboglobulin was detected in platelet-depleted plasma samples, indicative of platelet damage before plasma separation. CONCLUSION: Neither CsA nor tacrolimus induced active TGFbeta1 blood levels in liver transplant recipients during a follow up period of < or = 265 days. PMID- 10480423 TI - Analysis of variable region of T-cell receptor beta chain usage in the human anti porcine xenoresponse. AB - BACKGROUND: The human xenoreactive T-cell receptor (TCR) repertoire is not well documented. The aim of this study was to analyze the TCR repertoire in human anti porcine xenoresponses. METHODS: Peripheral blood lymphocytes (PBLs) from healthy donors were used to generate human T-cell lines against two different haplotypes of inbred Yucatan miniature swine (y/y and z/z). The variable region of TCR beta chain (Vbeta) gene usage was determined by fluorescence CDR3 spectrotyping. RESULTS: TCR Vbeta usage of an established human antiporcine T-cell line analyzed at weeks 5, 7, and 9 showed a sequential increase in Vbeta 1, 2, 6.2, 11, and 19 as compared to unprimed peripheral blood lymphocytes, whereas the usage of other Vbetas decreased. The selection of limited Vbeta genes correlated with the sequential increase in the specific lysis of the T-cell line, suggesting a non random clonal selection and expansion of T-cell clones that recognized porcine targets. Different Vbeta restriction was found using the same peripheral blood lymphocytes against a different haplotype of swine, indicating this selection of Vbeta gene was swine leukocyte antigen-dependent. CONCLUSIONS: There is restricted TCR Vbeta usage in the human anti-porcine response, suggesting that a limited number of xenogeneic epitopes are recognized by human T cells. The selection of particular TCR Vbeta clonotypes depends on the swine leukocyte antigen background. PMID- 10480424 TI - Repeat donor HLA-DR mismatches in renal transplantation: is the increased failure rate caused by noncytotoxic HLA-DR alloantibodies? AB - INTRODUCTION: Data from the UCLA/UNOS and Collaborative Transplant Studies Registries indicate that mismatched HLA-DR alloantigens expressed on a former donor renal allograft should not be repeated because of significantly poorer long term survival. METHODS: Retransplant candidates waiting for another renal allograft were screened for HLA class II alloantibodies (aAb) using direct complement-dependent cytotoxicity and several sensitive aAb binding assays. RESULTS: When screened by complement-dependent cytotoxicity, 46% of the patients were aAb negative. In contrast, using aAb binding assays, 90% of the patients had HLA-DR aAb specific for previous HLA-DR allograft mismatches. Most important, no directly cytotoxic HLA-DR antibody was detected in 9 of 27 patients. CONCLUSION: Our studies suggest that crossing the same HLA-DR mismatch in a subsequent transplant may result in poorer survival due to underlying donor-specific HLA-DR aAb. If confirmed in a retrospective study of retransplant patients, B cell donor cross-matches using antiglobulin complement-dependent cytotoxicity or flow cytometry would appear essential if this barrier were to be crossed. PMID- 10480425 TI - Clinical strategy for the management of acute humoral rejection in kidney transplantation. PMID- 10480426 TI - Neutrophil maturation and the role of retinoic acid. AB - Neutrophil maturation occurs in well defined morphological stages that correlate with the acquisition of molecular markers associated with neutrophil function. A variety of factors are known to play a role in terminal neutrophil maturation, including the vitamin A derivative, retinoic acid. Retinoic acid can directly modulate gene expression via binding to its nuclear receptors, which can, in turn, activate transcription of target genes. A role for retinoic acid during neutrophil maturation has been suggested from a variety of sources. Here we present a review of the mechanism of retinoic acid receptor action and the major evidence showing that normal retinoid signaling is required for neutrophil maturation. PMID- 10480427 TI - Listeria monocytogenes and recurrent mycobacterial infections in a child with complete interferon-gamma-receptor (IFNgammaR1) deficiency: mutational analysis and evaluation of therapeutic options. AB - We describe the history of a girl with interferon-gamma-receptor (IFNgammaR1) deficiency and studies performed to identify the molecular and clinical characteristics of this recently discovered disorder. This is the first report of a child from Northern Europe with IFNgammaR1 deficiency. The patient, now 7 years old, first presented with swelling and reddening at the Bacille Calmette-Guerin (BCG) vaccination site, swelling of lymph nodes, hepatomegaly, and an unusually severe varicella rash at the age of 4 months. At that time, she was diagnosed with BCG histiocytosis without typical granuloma formation and was treated with antituberculous agents. During the clinical course of her illness, several different types of atypical mycobacteria and (for the first time in an IFNgammaR1 deficient patient) Listeria monocytogenes were detected. Flow cytometric analysis showed that the patient's monocytes could not bind a monoclonal antibody specific for the IFNgamma-receptor. Our analysis of mRNA derived from the alpha-chain (IFNgammaR1) gene of this receptor revealed deletions of 173 bp and 4 bp in cDNA sequences originating from individual alleles. The 173 bp deletion was located between nucleotide positions 200 and 372, exactly matching those of exon 3, and the 4 bp deletion was located between nucleotide positions 561 and 564 of the coding region of the cDNA. Analysis of genomic DNA revealed the presence of a G to T transition at the 5'end of the splice consensus sequence of intron 3, which explains the absence of exon 3. The other allele carried the 4-base-pair deletion (ACTC) at nucleotide positions 15-18 of exon 5. Twelve months after an allo?geneic bone marrow transplantation, the patient had clinically improved. PMID- 10480428 TI - Analysis of T-cell defects in the specific immune response against acute lymphoblastic leukemia cells. AB - We previously showed that a specific antileukemia T-cytotoxic response is spontaneously elicited in acute lymphoblastic leukemia (ALL) patients and might contribute to host antileukemia defense, even though it is insufficient for tumor growth control. In this study, we report that multifactorial factors account for some of the acquired immune defects seen in ALL patients. In bone marrow of ALL patients, T cells do not express CD40L and CD25 markers, their apoptosis rate is increased, and a predominance of a CD4 cell subset expressing a Th2 phenotype is detected. A lack of expression of B7-1 molecules and other activation molecules is observed on all ALL blasts. These different parameters combined lead to in vivo dysfunction of T-cell proliferative and cytotoxic activity. PMID- 10480429 TI - Role of abnormal integrin-cytoskeletal interactions in impaired beta1 integrin function in chronic myelogenous leukemia hematopoietic progenitors. AB - Abnormal circulation and unregulated proliferation of chronic myelogenous leukemia (CML) progenitors is related, at least in part, to BCR/ABL induced abnormalities in beta1 integrin-mediated adhesion and signaling. The BCR/ABL oncogene has several potential interactions with cytoskeletal elements that are important for normal integrin signaling. In the present study, we evaluated whether abnormalities in beta1 integrin-cytoskeletal interactions were present in primary CML progenitors and contributed to defective integrin function. beta1 integrin-cytoskeletal interactions were studied in CML and normal CD34+ primary hematopoietic progenitors as well as BCR/ABL-transfected or mock-transfected M07e cells. In normal CD34+ progenitors, antibody-mediated cross-linking of beta1 integrins resulted in their redistribution into caps via a process requiring receptor-cytoskeletal interactions. CML CD34+ cells demonstrated significantly impaired beta1 integrin capping. This defect was related to the presence of the BCR/ABL gene, because capping also was impaired in BCR/ABL-transfected M07e cells. Defective receptor capping was not seen for non-integrin receptors. In addition, CML CD34- and M07eBCR/ABL cells also demonstrated increased actin polymerization and altered actin cytoskeletal organization. Further studies suggested that impaired beta1 integrin capping and defective integrin-mediated adhesion and proliferation inhibition in CML cells were related to abnormally enhanced integrin-cytoskeletal association and restricted receptor mobility. Finally, interferon alpha, which restores integrin-mediated adhesion and signaling in CML progenitors, also enhanced integrin capping in CD34+ cells. These studies suggest that p210BCR/ABL induces abnormal association of integrin receptors with the cytoskeleton and restricted receptor mobility and provide new insights into mechanisms underlying abnormal integrin function in CML progenitors. PMID- 10480430 TI - Leukemia patient-derived lymphoblastoid cell lines exhibit increased induction of leukemia-associated transcripts following high-dose irradiation. AB - Improvement in diagnostic cytogenetic techniques has led to the recognition of an increasing number of leukemia-associated chromosomal translocations and inversions. These genetic lesions frequently are associated with the disruption of putative transcription factors and the production of hybrid transcripts that are implicated in leukemogenesis. Epidemiologic evidence suggests that some, but not all, individuals with a history of gamma-irradiation exposure are at increased risk of developing chronic myeloid leukemia (CML). CML is characterized by the Philadelphia chromosome and transcription of the resulting hybrid BCR-ABL gene. Utilizing the leukemia-associated BCR-ABL p210 transcript as a marker, we sought differences in the induction of illegitimate genetic recombination following high-dose gamma-irradiation of karyotypically normal lymphoblastoid cell lines (LCL) derived from individuals with and without a history of myeloid leukemias. Six LCL [4 leukemia patient derived [2 acute myeloid leukemia and 2 CML] and 2 from normal individuals were analyzed with reverse transcriptase polymerase chain reaction for BCR-ABL under stringent conditions following exposure to 0, 50, or 100 Gy of LET gamma-irradiation delivered via a Varian linear accelerator at 4 MV. Transcripts identical to disease-associated b2a2 and b3a2 transcripts were detected both spontaneously (background illegitimate genetic recombination) and following gamma-irradiation. Background BCR-ABL positivity was demonstrable in 4 of the 6 LCL, with no significant difference in detection between leukemic- and nonleukemic-derived LCL. Overall, increasing gamma-irradiation dose resulted in an increased frequency of BCR-ABL transcript detection (0 Gy vs 50 Gy vs 100 Gy,p = 0.0023, Chi-square test). Within the leukemic- but not the nonleukemic-derived LCL there was significantly greater BCR ABL positivity after gamma-irradiation compared to unirradiated equivalents. Furthermore, the BCR-ABL positivity of both the AML- and CML-derived LCL after gamma-irradiation was significantly greater than that of the nonleukemic-derived LCL after gamma-irradiation. We speculate that this difference in the detection of illegitimate after gamma-irradiation recombination may be due to aberrant DNA double strand break repair mechanisms in individuals predisposed to the development of myeloid leukemias. PMID- 10480431 TI - CD95/Fas-triggered apoptosis of activated T lymphocytes is prevented by dendritic cells through a CD58-dependent mechanism. AB - T-cell apoptosis is a mechanism regulating T-cell homeostasis. Activation renders T cells susceptible to activation-induced cell death, a process mediated through CD95 ligand/CD95 (Apo-1/Fas) ligation. The aim of this study was to test whether antigen-presenting cells can inhibit CD95/Fas-triggered activation-induced cell death. Dendritic cells (DC), which are highly effective antigen-presenting cells, were generated in vitro from human peripheral blood monocytes by culture in granulocyte-macrophage colony-stimulating factor and interleukin 4. Subsequently, DC were cocultured with activated T cells and the effect of DC on CD95/Fas mediated apoptosis was determined. Coculture with increasing amounts of DC prevented CD95/Fas-triggered apoptosis in a dose-dependent fashion by inhibiting activation of caspase 8 and caspase 3. This protective effect of the DC on T-cell death could be blocked by 50% by adding an anti-CD58 antibody, whereas further addition of anti-CD80 (B7.1) and anti-CD86 (B7.2) led to an even more pronounced effect. Our findings suggest that DC can protect T cells from activation-induced cell death, with CD58 ligation playing a key role. PMID- 10480432 TI - Ectopic overexpression of c-mpl by retroviral-mediated gene transfer suppressed megakaryopoiesis but enhanced erythropoiesis in mice. AB - In this report, we tested whether ectopic overexpression of a cell surface receptor cDNA could be used to explore the physiological roles of that receptor. We generated c-mpl overexpressing animals by reconstituting mice with retroviral vector-transduced bone marrow (BM) cells. We observed that platelet counts in the c-mpl overexpressing mice failed to recover to normal levels and remained at <200 x 10(6)/mL post-transplantation, while platelet numbers in the control mice returned to > 800 x 10(6)/mL by 4 weeks post-transplantation. However, platelet counts in the c-mpl overexpressing mice could be stimulated to normal levels after administration of rhMGDF. No significant changes in peripheral leukocyte counts were observed, although the number of CFU-E, GM-CFC, and CFC-multi were reduced two- to threefold in the BM of the c-mpl overexpressing mice. In addition, enhanced erythropoiesis was observed in the c-mpl overexpressing mice. The mpl receptors on erythroid cells were functional as demonstrated by tyrosine phosphorylation of mpl receptor on RBC and by in vitro erythroid colony-formation in response to MGDF stimulation, respectively. These results suggested that ectopically expressed mpl receptors competed for ligand in vivo leading to an insufficient amount of circulating thrombopoietin (Tpo) for the development of megakaryocytic lineage. These results further suggest that, in addition to sequestering circulating Tpo, overexpression of the mpl receptor on erythroid progenitors may directly contribute to enhanced erythropoiesis in vivo. Our studies demonstrate that ectopic overexpression of a receptor by retroviral mediated gene transfer provides an approach to explore the biological roles of novel receptors. PMID- 10480433 TI - Functional differences between transplantable human hematopoietic stem cells from fetal liver, cord blood, and adult marrow. AB - The purpose of this study was to develop a simple assay for quantitating transplantable human lymphomyeloid stem cells (competitive repopulating units [CRU]) to enable comparison among the numbers and types of progeny generated in NOD/ SCID mice by such cells from different ontologic sources. Sub-lethally irradiated NOD/SCID mice were transplanted with varying numbers of CD34+ cell enriched suspensions of human fetal liver, cord blood, or adult marrow cells. The types and numbers of human cells present in the marrow of the mice were measured 6 to 8 weeks later using flow cytometry, in vitro progenitor assays, and secondary transplant endpoints. Frequencies of human CRU obtained by limiting dilution analysis of mice repopulated 6 to 8 weeks posttransplant were the same when the lymphoid and myeloid progeny of CRU were both detected by specific immunophenotypic endpoints as when in vitro myeloid progenitor assays were used to detect CRU myelopoietic activity. The average output per injected CRU of very primitive cells (CD34(+)CD38(-) cells, LTC-IC, and secondary CRU) was found to be highest for fetal liver CRU and progressively decreased (up to >100-fold) for ontologically older CRU. In contrast, the average output of mature cells was highest for cord blood CRU and lowest for fetal liver CRU, despite equivalent production of intermediate progenitors. Differences in the relative numbers of mature lymphoid, myeloid, and erythroid progeny produced by CRU from different ontologic sources were also seen. Finally, evidence of a transplantable human lymphoid-restricted cell present throughout ontogeny was obtained. A simpler and easier assay for enumerating transplantable human stem cells with lymphomyeloid reconstituting activity has been described, and its specificity and sensitivity validated. The use of this assay has revealed ontogeny-associated differences in a variety of functional attributes of human stem cells proliferating and differentiating in an in vivo, but xenogeneic, setting. PMID- 10480434 TI - Role of CD95/Fas and its ligand in the regulation of the growth of human CD34(++)CD38(-) fetal liver cells. AB - The functional significance of CD95/Fas expressed by candidate hematopoietic stem cells (HSCs) from human fetal liver was studied by testing the effect of agonistic anti-CD95 monoclonal antibody (mAb) CH-11 and soluble CD95 ligand (sCD95L) on the growth of CD34(++)CD38(-)lineage cells in vitro. Candidate fetal HSCs exhibited a dose-dependent proliferative response to CH-11 as well as to sCD95L when combined with kit ligand (KL) + interleukin 3 (IL-3) under serum deprived culture conditions. CH-11 mAb increased, in a synergistic fashion, the number of myeloid colony-forming unit culture (CFU-C) generated by candidate HSCs in liquid cultures with the cytokine combinations KL + IL-3, KL + granulocytemacrophage colony-stimulating factor, and KL + IL-6. CH-11 mAb and sCD95L also enhanced erythropoiesis supported by KL + IL-3 + erythropoietin (Epo). Furthermore, sCD95L was able to increase the number of megakaryocytes, granulocytes, and CD34- cells generated in the presence of KL + IL-3 + Epo + thrombopoietin. An analysis performed using Western blotting revealed that the membrane-bound CD95L (mCD95L) was expressed by both immature (total CD34+/++) and mature (CD34-) hematopoietic lin(-) FL cells. Among the CD34(++)lin(-)cells, both the freshly isolated CD38+ and CD38 subsets as well as CD95+ and CD95- cells constitutively expressed mCD95L, demonstrating that the CD95/CD95L system represents a paracrine and potentially autocrine regulator of early hematopoiesis. To study the role of the endogenously produced CD95L, we determined the effects of a neutralizing anti-CD95L NOK-1 on the growth of candidate HSCs. By blocking the endogenous CD95L with NOK-1 mAb, we observed an increase in CFU-C generated by candidate HSCs. We conclude that the endogenous CD95L has an inhibitory effect on fetal candidate HSCs, which can be blocked by sCD95L and CH-11 mAb. PMID- 10480435 TI - Intrinsic potential of phenotypically defined human hemopoietic stem cells to self-renew in short-term in vitro cultures. AB - In search for culture conditions that will facilitate hemopoietic stem cell (HSC) replication while preserving their primitive properties, we have made use of a multi-parameter FACS assay to define HSCs on basis of their phenotypic characteristics, i.e., CD34++CD33,38,71(-). Bone marrow and umbilical cord blood samples of CD34(+) cells from 31 donors were loaded with the membrane dye PKH26 and each exposed to various culture conditions for 6 days. The cells that retained the primitive CD34(++)CD33,38,71(-) phenotype were analysed for the number of cell replications they underwent, by measuring loss of PKH26 fluorescence after 6 days. A most striking observation was the large inter-sample variation in the proliferative response of cells that retained the CD34(++)CD33,38,71(-) phenotype. In general, samples could be characterised as either good- or poorly-replicating, according to the proliferation property of their CD34(++)CD33,38,71(-) subset. In comparison to this 'intrinsic' potential, the effects of the applied growth stimuli on CD34(++)CD33,38,71(-) cell replication were negligible. In contrast, the overall recovery of the CD34(++)CD33,38,71(-) cells was clearly dependent on the culture stimuli. Of the various conditions tested, serum-free cultures with pre-established stroma maintained the cells with this primitive phenotype most effectively. In cultures supplemented with various combinations of recombinant HGFs, HSC differentiation prevailed. These findings with phenotypically defined HSCs should assist in the design of systems for expansion and ex vivo gene therapy of early hemopoietic cells. PMID- 10480436 TI - Fas ligand promotes cell survival of immature human bone marrow CD34+CD38- hematopoietic progenitor cells by suppressing apoptosis. AB - Fas (CD95, APO-1) is a member of the TNF receptor family, and engagement of Fas by its ligand, Fas ligand (FasL), can induce apoptotic death of Fas expressing cells. Signaling through Fas has previously been shown to induce apoptosis of CD34+ human hematopoietic progenitor cells after exposure to IFN-gamma or TFN alpha. In contrast, we found that FasL promoted a significantly increased viability of primitive CD34+CD38- cells. Thus, incubation with FasL for 48 hours reduced cell death from 46 to 29% compared to cells cultured in medium alone as measured by propidium iodide (PI) incorporation (n = 8, p < 0.02). Inhibition of apoptosis was confirmed by morphological analysis and by the Nicoletti technique. Furthermore, by using a delayed addition assay at the single cell level we found that sFasL treatment had a direct viability-promoting effect on CD34(+)CD38(-) cells. The effect of sFasL was completely blocked by NOK-1, a neutralizing mAb against FasL. In agreement with previous reports, FasL alone slightly increased cell death of more mature CD34(-)CD38+ cells, indicating an interesting shift in the responsiveness to FasL during early hematopoiesis. PMID- 10480437 TI - Stromal damage as consequence of high-dose chemo/radiotherapy in bone marrow transplant recipients. AB - Bone marrow transplant (BMT) relies on the engraftment of donor hemopoietic precursors in the host marrow space. Colony forming units-fibroblasts (CFU-f), the precursor compartment for the osteogenic lineage, are essential to hemopoietic stem cell survival, proliferation and differentiation. We have studied CFU-f in donors (aged 5 months to 62 years) and in patients who had received allogeneic BMT (aged 2 months to 63 years). In donor marrows we found an inverse correlation between CFU-f frequency and age. In BMT recipients CFU-f frequencies were reduced by 60%-90% (p < 0.05) and the numbers did not recover up to 12 years after transplant. Stromal reconstitution to normal levels was found only in patients < 5 years old. In all patients studied CFU-f post-BMT were of host origin. Patients with low CFU-f levels displayed also a decreased bone mineral density (p < 0.05) and significantly reduced levels of long-term culture initiating cells (LTC-IC) (p < 0.05). Our study demonstrates that the marrow stromal microenvironment is seriously and irreversibly damaged after BMT. Donor cells do not contribute to reconstitute the marrow microenvironment, whose residual CFU-fs remain of host origin. PMID- 10480439 TI - Can physicians always explain the results of clinical trials? A case study of amlodipine in heart failure. PMID- 10480438 TI - Correction of peripheral lysosomal accumulation in mice with aspartylglucosaminuria by bone marrow transplantation. AB - OBJECTIVE: Bone marrow transplantation has been shown to alleviate symptoms outside the CNS in many lysosomal storage diseases depending on the type and stage of the disease, but the effect on neurological symptoms is variable or still unclear. Aspartylglucosaminuria (AGU) is a lysosomal storage disease characterized by mental retardation, recurrent infections in childhood, hepatosplenomegaly and coarse facial features. Vacuolized storage lysosomes are found in all tissues of patients and uncleaved enzyme substrate is excreted in the urine. The recently generated AGU mouse model closely mimicks the human disease and serves as a good model to study the efficiency of bone marrow transplantation in this disease. METHODS: Eight-week-old AGU mice were lethally irradiated and transplanted with bone marrow from normal donors. The AGA enzyme activity was measured in the liver and the brain and the degree of correction of tissue pathology was analyzed by light and electron microscopy. Reverse bone marrow transplantation (AGU bone marrow to wild-type mice) was also performed. RESULTS: Six months after transplantation the AGA enzyme activity was 13% of normal in the liver, but only 3% in the brain. Tissue pathology was reversed in the liver and the spleen, but not in the brain and the kidney. The urinary excretion of enzyme substrate was diminished but still detectable. No storage vacuoles were found in the tissues after reverse transplantation, but subtle excretion of uncleaved substrate was detected in the urine. CONCLUSION: Liver and spleen pathology of AGU was corrected by bone marrow transplantation, but there was no effect on lysosomal accumulation in the CNS and in the kidneys. PMID- 10480440 TI - Amlodipine therapy in congestive heart failure: hemodynamic and neurohormonal effects at rest and after treadmill exercise. AB - This study examined the acute effects of amlodipine treatment on left ventricular pump function, systemic hemodynamics, neurohormonal status, and regional blood flow distribution in an animal model of congestive heart failure (CHF), both at rest and with treadmill exercise. A total of 14 pigs were studied under control conditions and after the development of pacing-induced CHF (240 beats per minute, 3 weeks, n = 7) or with CHF and acute amlodipine treatment for the last 3 days of pacing (1.5 mg/kg per day, n = 7). Under resting conditions, left ventricular stroke volume (mL) was reduced with CHF compared with the normal state (15+/-2 vs. 31+/-1, p<0.05) and increased with amlodipine treatment (23+/-4, p<0.05). At rest, systemic vascular resistance increased with CHF compared with the normal state (3,078+/-295 vs. 2,131+/-120 dyne x s cm(-5), p<0.05) and was reduced after amlodipine treatment (2,472+/-355 dyne x s cm(-5), p<0.05). With exercise, left ventricular stroke volume remained lower and systemic vascular resistance higher in the CHF group, but was normalized with amlodipine treatment. With exercise, left ventricular myocardial blood flow increased from resting values, but was reduced from the normal state with CHF (normal: 1.69+/-0.12 to 7.62+/-0.74 mL/min per gram vs. CHF: 1.26+/-0.12 to 4.77+/-0.45 mL/min per gram, both p<0.05) and was normalized with acute amlodipine treatment (1.99+/-0.35 to 6.29+/-1.23 mL/min per gram). Resting plasma norepinephrine was increased by >5-fold in the CHF group at rest and was not affected by amlodipine treatment. However, with exercise, amlodipine treatment blunted the increase in plasma norepinephrine by >50% when compared with untreated CHF values. Resting plasma endothelin levels increased with CHF compared with the normal state (10.9+/-0.9 vs. 2.8+/-0.4 fmol/mL, p<0.05) and was reduced with amlodipine treatment (7.5+/-1.5 fmol/mL, p<0.5). In other vascular beds, acute amlodipine treatment with CHF improved pulmonary and renal blood flow both at rest and with exercise; however, there were no effects observed on skeletal muscle blood flow. With the development of CHF, acute amlodipine treatment does not negatively influence left ventricular pump function, but rather may provide favorable hemodynamic and neurohormonal effects. PMID- 10480441 TI - Antioxidant properties of calcium antagonists related to membrane biophysical interactions. AB - The antioxidant activities of representative calcium antagonists, including amlodipine, verapamil, and diltiazem, were measured in hepatic microsomal membranes by the Fe-catalyzed, hydroxyl radical-producing system (dihydroxyfumarate + Fe3+) and assessed by malondialdehyde (MDA) formation. Despite the absence of L-type calcium channels in this membrane preparation, the calcium antagonists showed dose-dependent antioxidant activity. The biophysical mechanism for calcium-antagonist antioxidant activity was evaluated using radioligand binding assays, high-resolution differential scanning calorimetry, and small-angle x-ray diffraction approaches. These analyses demonstrated that calcium-antagonist antioxidant potency correlated directly with the compounds' relative affinity for the membrane lipid bilayer and ability to modulate membrane thermodynamic properties (amlodipine >> verapamil > diltiazem). The charged 1,4 dihydropyridine calcium antagonist, amlodipine, had the highest affinity for the membrane lipid bilayer (Kp>10(4)) and produced the largest changes in membrane thermodynamic properties, including a reduction in thermal phase transition temperature (-11%), enthalpy (-14%), and cooperative unit size (-59%), relative to control phosphatidylcholine liposomes. Electron density profiles generated from x-ray diffraction data demonstrated that amlodipine effected a broad and dose-dependent increase in molecular volume associated with the membrane hydrocarbon core. These data indicate that lipophilic calcium antagonists inhibit lipid peroxidation in cellular membranes as a result of modulating physicochemical properties of the membrane lipid bilayer, independently of calcium channel inhibition. Amlodipine had the most potent antioxidant activity as a result of distinct biophysical interactions with the membrane lipid bilayer. The nonreceptor-mediated antioxidant activity of calcium antagonists may contribute to cytoprotective mechanisms of action in cardiovascular diseases. PMID- 10480442 TI - Potential role of the microvasculature in progression of heart failure. AB - After cardiac injury, there are changes in the cardiac myocyte morphology, function, matrix, and molecular gene expression. These all play an important role in remodeling of the injured heart, contributing to the progression toward heart failure. The role of the microvasculature in the progression toward heart failure is less well characterized. However, laboratory studies have established that there are important interactions between the microvascular endothelium and the myocyte. Furthermore, in a multitude of animal models of heart failure and cardiomyopathy, there is always an association with microvascular abnormalities. Reversal of these abnormalities is also associated with improvement in the cardiomyopathy. Major mediators that likely play an important role in the microvasculature include endothelin and nitric oxide. These are elaborated by both endothelium and myocyte compartments of the myocardium. Preliminary clinical studies already demonstrate that microvascular ischemia may have prognostic power in patients with nonischemic dilated cardiomyopathy. Results from these studies showed a reduction in mortality from treatment with amlodipine, suggesting a possible benefit based on changes in the microvasculature. PMID- 10480443 TI - Amlodipine promotes kinin-mediated nitric oxide production in coronary microvessels of failing human hearts. AB - Recently, we found that amlodipine can release nitric oxide (NO) from canine coronary microvessels, which raises the question of whether amlodipine can also promote coronary NO production in failing human hearts. The goal of this study was to define the effect of amlodipine on NO production in failing human hearts and to determine the role of kinins in the control of NO production induced by amlodipine. Six explanted human hearts with end-stage heart failure were obtained immediately at transplant surgery. Coronary microvessels were isolated as previously described, and nitrite, the stable metabolite of NO in aqueous solution, was measured using the Griess Reaction. Amlodipine (10(-10) to 10(-5) mol/L) significantly increased nitrite production in coronary microvessels in a dose-dependent manner. The increase in nitrite in response to the highest dose of amlodipine (79%) was similar in magnitude to either that of the angiotensin converting enzyme inhibitor ramiprilat (74%) or the neutral endopeptidase inhibitors phosphoramidon (61%) and thiorphan (72%). Interestingly, the increase in nitrite production induced by amlodipine was entirely abolished by N(omega) nitro-L-arginine methyl ester and also HOE-140 (a bradykinin-2 antagonist) and dichloroisocoumarin (a serine protease inhibitor that blocks kallikrein activity). These results indicate that amlodipine can promote coronary NO production in failing human hearts and that this effect is dependent on a kinin mediated mechanism. PMID- 10480444 TI - Tocotrienol: a review of its therapeutic potential. AB - OBJECTIVES: To summarize new knowledge surrounding the physiological activity of tocotrienol, a natural analogue of tocopherol. RESULTS: The biological activity of vitamin E has generally been associated with its well-defined antioxidant property, specifically against lipid peroxidation in biological membranes. In the vitamin E group, alpha-tocopherol is considered to be the most active form. However, recent research has suggested tocotrienol to be a better antioxidant. Moreover, tocotrienol has been shown to possess novel hypocholesterolemic effects together with an ability to reduce the atherogenic apolipoprotein B and lipoprotein(a) plasma levels. In addition, tocotrienol has been suggested to have an anti-thrombotic and anti-tumor effect indicating that tocotrienol may serve as an effective agent in the prevention and/or treatment of cardiovascular disease and cancer. CONCLUSION: The physiological activities of tocotrienol suggest it to be superior than alpha-tocopherol in many situations. Hence, the role of tocotrienol in the prevention of cardiovascular disease and cancer may have significant clinical implications. Additional studies on its mechanism of action, as well as, long-term intervention studies, are needed to clarify its function. From the pharmacological point-of-view, the current formulation of vitamin E supplements, which is comprised mainly of alpha-tocopherol, may be questionable. PMID- 10480445 TI - Regulators of apoptosis in human breast cancer. AB - OBJECTIVES: Apoptosis or programmed cell death represents a mechanism by which tumor cells with DNA damage can be deleted. Bcl-2 and p53 gene products have been both linked to apoptosis. Bcl-2 plays a role as an inhibitor of apoptosis that may extend the viability of cells containing genetic alterations and facilitate tumor progression. Mutant p53 has a similar effect. The purpose of this study was to investigate expression of bcl-2 in 70 malignant and 30 benign breast lesions using different methods (enzyme immunoassay, immunodot blot, Western blot) and to compare it with the established clinicopathological prognostic factors (age, tumor size, type, grade, lymph node status) and some molecular genetic markers in breast cancer. RESULTS: bcl-2 and mutant p53 were highly expressed in breast cancer than benign breast lesions and aneuploidy was more frequently detected in malignant breast samples. No correlation could be observed between bcl-2 expression and node status, tumor size, differentiation, type, age at excision or mutant p53 expression. However, a strong positive associations were seen between bcl-2 and estrogen receptors (ER), DNA aneuploidy. Eighty-five percent of bcl-2 positive tumors were ER positive and 65% were aneuploid, while in bcl-2 negative tumors only 28% were ER positive and 37% were aneuploid. CONCLUSIONS: The association seen between bcl-2 and ER raises the possibility that bcl-2 is an ER regulated gene which suggests a potential important role for bcl-2 as a modulator of response to hormonal therapy in breast cancer. Monitoring hormonal therapy can easily be done by bcl-2 quantitative EIA method. PMID- 10480446 TI - Rapid and accurate determination of (CAG)n repeats in the androgen receptor gene using polymerase chain reaction and automated fragment analysis. AB - OBJECTIVES: To develop and evaluate a new method for determination of the CAG repeat length in Exon 1 of the androgen receptor gene. DESIGN AND METHODS: The method is based on PCR amplification of a DNA region encompassing the repeats and analysis of the length of the PCR product on a sequencing gel. One of the PCR primers was labeled with Cy5.5 fluorescent dye to facilitate detection after laser excitation. We used a fully automated system for electrophoretic separation of the PCR product and accurate sizing of the length of the PCR product using fragment analysis. RESULTS: The major advantages of the new technique are its simplicity, speed, accuracy, and reproducibility. Analysis of the CAG repeats in genomic DNAs from 18 males indicated that they were all hemizygous with a mean CAG repeat number of 22 (range 20-30 repeats). Among 60 DNAs from females, 16 were homozygous and 44 were heterozygous. The repeat length ranged from 17-30 with a mean of 22. In both males and females, the distribution of CAG repeats was bimodal. CONCLUSION: We anticipate that this improved method for CAG repeat analysis will find applications in clinical studies involving prostate and breast cancer patients. PMID- 10480447 TI - Increased bcl-2 expression is associated with primary resistance to chemotherapy in human epithelial ovarian cancer. AB - OBJECTIVE: bcl-2, an anti-apoptotic factor, has a role in the pathogenesis of ovarian cancer as well as in resistance to chemotherapy. DESIGN AND METHODS: 20 benign, and 26 malignant epithelial ovarian tissues were analyzed for bcl-2 protein and mutant p53 by enzyme-immunoassay (EIA). Flowcytometric analysis was also performed. Patients of malignant group were followed up to monitor overall survival and primary resistance to chemotherapy. RESULTS: bcl-2 was significantly higher in malignant group than benign group (p < 0.001). A cutoff value was determined for bcl-2 (63.8 kU/g protein). At this cutoff, sensitivity is 80.7%, and specificity is 85%. Using chi square analysis, a significant correlation was found between bcl-2 and FIGO stage (p = 0.01), overall survival (p = 0.01), as well as primary resistance to chemotherapy (p = 0.03). By correlation coefficient analysis the relation between bcl-2 and synthetic phase fraction was highly significant (p = 0.002). Bcl-2, p53, and FIGO stage were significantly correlated to poor survival (p = 0.01) in univariate analysis. However, in multivariate analysis, only FIGO stage, and p53 were independent risk factors. CONCLUSION: EIA could be a useful tool for investigating the prognostic value of bcl-2, and its possible prediction of platinum resistance in epithelial ovarian cancer. This might help in selecting patients for future anti-bcl-2 therapy. PMID- 10480448 TI - Homogeneous HDL-cholesterol assay versus ultracentrifugation/dextran sulfate-Mg2+ precipitation and dextran sulfate-Mg2+ precipitation in healthy population and in hemodialysis patients. AB - OBJECTIVES: To evaluate the analytical performance of a new homogeneous HDL cholesterol assay (Olympus Diagnostica). To investigate possibly discrepant results in chronic hemodialysis patients who commonly exhibit quantitative and qualitative lipoprotein abnormalities, responsible for atherogenic complications in these patients. DESIGN AND METHODS: Serum samples were collected from 50 healthy subjects and 65 chronic hemodialysis patients. HDL-C levels measured by the homogeneous assay were compared with the routine dextran sulfate-Mg2+ precipitation method and the ultracentrifugation/dextran sulfate-Mg2+ precipitation as reference method. RESULTS: The homogeneous assay was linear up to at least 220 mg/dL. The analytical precision was estimated with three different sets of commercial controls and one set of human pooled serum control. The within-day CV ranged between 1.7% and 3.8% and the between-day CV ranged between 1.0% and 2.3%. HDL-C values in both populations correlated highly with the dextran sulfate-Mg2+ precipitation method and the ultracentrifugation/dextran sulfate-Mg2+ precipitation method (r > or = 0.96, bias between -0.9 and 2.3 mg/dL). Lipemia up to triglyceride concentration of 600 mg/dL did not alter the HDL-C value. CONCLUSIONS: The homogeneous assay for HDL-C (Olympus) uses much less sample, is accurate and convenient to handle, and allows full automation. The test should considerably facilitate the screening of individuals at an increased risk of cardiovascular disease, including hemodialysis patients. PMID- 10480449 TI - DNA aneuploidy, S-phase fraction and nuclear p53 positivity in non-small cell lung carcinoma. AB - OBJECTIVES: Surgical resection currently offers the best option for managing non small cell lung carcinoma (NSCLC) but its efficiency is limited by subsequent tumor recurrence. We evaluated whether flow cytometric study and the p53 gene staining pattern may be useful in this respect. DESIGN AND METHODS: We took biopsy samples of 40 patients with operable NSCLC to study the frequency of aneuploidy, proliferative activity, and alterations in the p53 tumor suppressor gene and compared them with clinicopathological parameters such as age, gender, smoking, histological type, tumor size, and degree of differentiation. To study DNA content, the nuclei were analyzed by flow cytometry using a FACS flow cytometer (Becton-Dickinson) equipped with an argon ion laser, with a propidium iodide excitation of 488 nm. We used the immunohistochemical technique for p53 analysis in samples of paraffin-embedded tissue corresponding to the same patients from whom fresh tissue was taken. RESULTS: Nuclear p53 staining was detected in 66.7% of the samples; 69.4% of the cases revealed aneuploid DNA histograms and 59% presented with an S phase fraction of more than 12%. Comparison with clinicopathological parameters showed that p53 protein was associated significantly with histological classification (p = 0.04), gender (p = 0.01), and smoking (p = 0.04). CONCLUSIONS: Immunodetection of p53 overexpression and DNA ploidy in the bronchial epithelium may be a useful tool in any future multifactorial analysis in such tumors for identifying previous lesions that may progress to malignancy. PMID- 10480450 TI - Correlation between the administered dose of DHEA and serum levels of DHEA and DHEA-S in human volunteers: analysis of published data. AB - OBJECTIVES: Studies from both experimental animals and humans suggest that administration of exogenous DHEA may have beneficial endocrine-metabolic, immunologic and neurologic effects. Several groups have administered DHEA to humans, but to the best of our knowledge, no one at this point has published a summary of the relationship between the administered dose of DHEA and the serum levels of steroids attained. DESIGN: We summarize the relationship between the administered dose of DHEA and the resulting serum level of DHEA and DHEA-S, in humans, from 18 published articles. RESULTS: Serum levels of DHEA and DHEA-S increase with increasing doses. Doses above 50 mg/day result in levels that are at or above the upper limit of normal for healthy young adults. At doses above 300 mg/day the increment of serum DHEA and DHEA-S appears to reach a plateau. CONCLUSIONS: Those wanting to use supplemental DHEA might consider that doses of 300 mg/day are maximal; they clearly result in supraphysiologic concentrations and above this level doses may have increased side effects without significantly increasing the effective level of serum hormone. PMID- 10480451 TI - Effects of propylthiouracil, propranolol, and vitamin E on lipid peroxidation and antioxidant status in hyperthyroid patients. AB - OBJECTIVE: The purpose of this study was to examine lipid peroxidation and antioxidant states during hyperthyroidism states and after given different treatments. DESIGN AND METHODS: We examined 44 hyperthyroid patients and 19 euthyroid healthy controls. Patients were divided into three groups according to the treatment: Propylthiouracil (PTU) group, PTU + propranolol (PRP) group, PTU + PRP + vitamin E (vitE) group. RESULTS: In the hyperthyroid patients plasma malondialdehyde (MDA) levels were significantly high as compared to the control group (p < 0,001). There was a significant decrease in the MDA levels post treatment (p < 0.001 in the PTU + PRP group and PTU + PRP + vitE group, p < 0.01 in the PTU group). In the hyperthyroidism, blood reduced glutathione (GSH) levels were lower, erythrocyte superoxide dismutase (SOD) and catalase (CAT) activities were higher than in the control group, but these changes were not significant. Post-treatment in each of the three groups the GSH levels were increased significantly as compared to the pretreatment levels (p < 0.001). There was significant decrease in the SOD activity post treatment (p < 0.01 in all three groups). Post-treatment CAT activity was decreased (p < 0.05 in the PTU group, p < 0.001 in the other two groups). The erythrocyte glutathione peroxidase (Gpx) activity was lower significantly in the hyperthyroidism as compared to the control group (p < 0.001). Post-treatment, in the three groups Gpx activity increased significantly as compared to the pretreatment levels (p < 0.05 in the PTU group, p < 0.001 in the PTU + PRP group and PTU + PRP + vitE group). CONCLUSION: We considered that giving all three treatments would be useful to the prevention of oxidative stress in the hyperthyroidism states. PMID- 10480452 TI - Evaluation of oxidative stress parameters in blood of patients with laryngeal carcinoma. AB - OBJECTIVES: In this study, plasma lipid peroxidation as assessed by thiobarbituric acid reactive substances and erythrocyte antioxidant status markers namely CuZn superoxide dismutase, glutathione peroxidase, glutathione and plasma levels of vitamin C and E were investigated in 20 patients with larygneal carcinoma and 15 healthy controls. DESIGN AND METHODS: Lipid peroxidation was observed to be significantly higher (0.01 > p > 0.001) in the larynx carcinoma group in comparison to the healthy controls. Both stage I + 11 and stage III carcinoma patients were observed to have significantly higher thiobarbituric acid reactive substances than the control group. A significant difference was found in plasma vitamin E level between the control group and stage I + 11 and stage III carcinoma patients (p < 0.01, 0.05 > p > 0.02, respectively). RESULTS: Our findings reveal the presence of increased lipooxidative damage in laryngeal carcinoma patients, but no change with respect to the endogenous antioxidant components-GSH, GSH Px, and CuZn SOD. PMID- 10480453 TI - Androgen imbalance in premenopausal women with benign breast disease and breast cancer. AB - OBJECTIVE: The alteration of steroid hormonal status in premenopausal breast disease (benign and malignant) were investigated by comparing the urinary profile of androgens and corticoids. METHODS: The urinary concentrations of 25 androgens and corticoids were quantitatively determined by a gas chromatographymass spectrometry system in patients with benign breast disease (35 cases, 20-54 years), breast cancer (34, 27-54), and healthy controls of similar age (25, 22 51). RESULTS: In premenopausal patients with breast cancer, a significantly lower rate of excretion of 11-deoxy-17-ketosteroids and their metabolites was found in comparison with normal females. These levels were also inversely associated with benign breast disease. No significant differences were found between the three groups for the concentration of 11-oxy-17-ketosteroids, 17-hydroxy-corticoids and their metabolites. The urinary ratio of adrenal androgen metabolites to cortisol metabolites [(11-DOKS & M)/11-OKS] declined in the order of normal female control (4.04 +/- 0.72; mean +/- SD), breast benign mass (2.29 +/- 0.42) and breast cancer (0.94 +/- 0.27). CONCLUSION: Our data suggest that the hormonal imbalance of androgen deficiency and/or corticoid sufficiency is closely associated with the benign and malignant conditions of premenopausal breast disease and the ratio of (11-DOKS & M)/11-OKS may be an effective discriminant factor of these groups. PMID- 10480454 TI - Monthly intra-individual variation in lipids over a 1-year period in 22 normal subjects. AB - OBJECTIVES: Previous reports on biological variation in lipids differ widely in the time interval between sampling, the number of samples analyzed per patient and the total study period. The present investigation was carried out to determine monthly intra-individual variation in lipids over 1 year and to establish whether there was a consistent change in lipid values over the summer months. The importance of taking this variation into consideration during the assessment of risk of coronary heart disease (CHD) was also examined. DESIGN AND METHODS: Cholesterol, triglycerides, HDL, apo A1, and apo B were measured at monthly intervals for 12 months in 22 healthy, free-living volunteers (11 females, 11 males) by standardized methods. RESULTS: When compared to analytical variation, biological variation was the dominant component of the intra individual changes observed during the 1-year study period. As expected, triglycerides showed the greatest biological variation; the ratio of biological/analytical variation was 33.1. Much smaller ratios were observed for the other lipids measured in this study with values ranging from 4.2 to 6.8. Different subjects attained their maximum and minimum values in virtually every month of the year. There were significant reductions in cholesterol, HDL, LDL, and apo A1 in the summer months while triglycerides showed a non-significant increase and apo B a non-significant decrease during this period. CONCLUSIONS: All the analytes showed considerable intra-individual variation. It is, therefore, important to measure lipids sequentially over several weeks to arrive at an average value for risk stratification for CHD. PMID- 10480455 TI - Fluorescence probe (BCECF) loading efficiency in human platelets depends on cell concentration: application to pHi measurements. PMID- 10480456 TI - Pasteurellaceae isolated from tonsillar samples of commercially-reared American bison (Bison bison). AB - As commercial producers of American bison (Bison bison) become more numerous, concerns relative to bison health management increase. Since loss due to respiratory disease associated with Pasteurella and related Pasteurellaceae is a major concern for cattle producers, a study was conducted to determine what types of Pasteurellaceae are carried by bison to evaluate the potential of pneumonic pasteurellosis in bison herds where management practices are comparable to those used for cattle. Tonsillar biopsies, collected in May (n = 29) and August (n = 25) 1997 from 24- to 30-month-old bison bulls, at the time of slaughter were cultured for Pasteurellaceae. Pasteurella spp. were isolated from all the samples collected in May. These included isolates identified as P. haemolytica, trehalosi, testudinis, and multocida subsp. multocida a and multocida b. Actinobacillus spp. and Haemophilus somnus were also isolated from some samples. Pasteurella spp., haemolytica, trehalosi, and multocida subsp. multocida a, multocida b and septica, plus 2 nonspeciated indole-positive biotypes, U2 and U16, were isolated from the second group of tonsil samples. Most of these organisms, including P. haemolytica, P. multocida subsp., and H. somnus are associated with disease in domestic livestock and should be regarded as potential pathogens for bison, particularly in animals which become stressed by management practices commonly used with cattle such as herding, crowding, and shipping. PMID- 10480457 TI - Characterization of putative Haemophilus somnus isolates from tonsils of American bison (Bison bison). AB - Three Haemophilus somnus isolates (2a, 3a, and 27b) and one H. somnus-like (13b) isolate from tonsils of commercially reared American bison were compared with 2 known H. somnus isolates from cattle, namely, 2336, shown to cause respiratory disease, and 129Pt, from the prepuce of an asymptomatic bull. All H. somnus isolates, but not the H. somnus-like isolate, required CO2 for growth. Biochemical utilization profiles were identical for bison and bovine H. somnus isolates with the exception of alpha-fucosidase production by isolate 3a. Isolate 27a varied from 2a, 2336 and 129Pt by hemolysis of bovine erythrocytes. Isolate 13b hemolyzed sheep but not bovine or bison erythrocytes and varied from other isolates in biochemical utilization tests. Outer membrane protein profiles of 2a, 3a and 27a were almost identical with those of bovine isolate 2336 and similar to that of 129Pt, but quite different from that of 13b. Western blots of bison isolates were similar to that of the virulent bovine 2336 isolate, including detection of high molecular mass antigens above 100 kDa and the 76 kDa antigens associated with bovine IgG2 Fc binding characteristic of virulent strains, as well as antigens of approximately 78, 60 and 40 kDa. Producers and veterinarians should be aware that H. somnus may be carried by bison and may have potential for causing diseases in bison similar to those described in cattle and sheep. PMID- 10480458 TI - Genomic relatedness among Actinobacillus pleuropneumoniae field strains of sterotypes 1 and 5 isolated from healthy and diseased pigs. AB - Forty-four Actinobacillus pleuropneumoniae isolates recovered from both healthy and diseased pigs were characterized by random amplified polymorphic DNA analysis (RAPD), pulsed field gel electrophoresis (PFGE) and apx toxin gene typing. Nine RAPD types and 14 PFGE patterns were identified. No common RAPD or PFGE patterns were found between strains of serotype 1 and those of serotype 5. The RAPD analysis indicated that the 15 serotype 1 strains isolated from diseased pigs were assigned to 4 RAPD types, with 66% of strains characterized by the same RAPD type. By contrast, the 5 strains of serotype 1 isolated from healthy carriers were dispersed in 4 RAPD types. These data suggest that the diversity of strains isolated from healthy pigs could be higher than that of strains recovered from diseased pigs. In addition, all serotype 5 strains exhibited a unique RAPD type. Unlike RAPD, PFGE analysis allowed discrimination among isolates of serotype 1 and among those of serotype 5. All but 3 isolates showed the same apx genotype as their respective serotype reference strain. These data indicate that RAPD analysis is a valuable rapid tool for routine subtyping of strains of serotype 1. For strains of serotype 5, a combination of several typing methods, such as PFGE and apx gene typing, is needed to provide useful information on the molecular epidemiology of swine pleuropneumonia. PMID- 10480459 TI - Virulence markers in Shiga toxin-producing Escherichia coli isolated from cattle. AB - This study identified potential virulence markers in 93 eae-positive and 179 eae negative Shiga toxin-producing Escherichia coli (STEC), isolated from a random sampling of healthy cattle in southwestern Ontario. PCR amplification was used to identify genes for enterohemorrhagic E. coli (EHEC)-hemolysin, the EAF plasmid, and bundle-forming pili (Bfp); adherence to HEp-2 cells and to bovine colonocytes, and the fluorescent actin staining (FAS) test were used to characterize interaction of the bacteria with epithelial cells. The EHEC hemolysin sequences were detected in 98% of eae-positive isolates compared with 34% of eae-negative isolates. All isolates were negative for EAF and bfp sequences. There was 100% correlation between localized adherence (LA) to HEp-2 cells and the FAS test. Forty-eight (52%) of the eae-positive isolates were LA/FAS-positive, whereas none of the 179 eae-negative isolates was positive in either test. Among the eae-negative isolates, 20 (11%) showed diffuse adherence and 5 (2.8%) showed enteroaggregative adherence to HEp-2 cells. Seventy-three percent of the eae-positive isolates adhered to bovine colonocytes, whereas only 26% of 120 eae-negative isolates that were tested adhered. All 13 O157:H7 isolates were positive for eae and EHEC-hemolysin gene sequences, LA/FAS, and adherence to bovine colonocytes. It is concluded that possession of genes for eae and EHEC hemolysin is correlated with the serotype of STEC, that production of EHEC hemolysin was highly correlated with serotypes implicated in human disease, and that none of the potential markers that were examined can be used to predict the potential virulence of an isolate. PMID- 10480460 TI - An epidemiologic study of disease in 32 registered Holstein dairy herds in British Columbia. AB - Data recorded in a herd health management system were obtained from 32 registered Holstein dairy herds from British Columbia. Frequencies of disease were described, and the effect of herd, age, year, season, and the interrelationships between diseases within a lactation on the occurrence of disease were evaluated. Lactational incidence rates were computed for diseases with a short period of risk (ie, udder edema, milk fever, retained placenta, metritis, displaced abomasum, and ketosis), whereas for diseases with a longer period of risk (ie, cystic ovaries, mastitis and stable footrot), incidence densities were calculated. Overall, the disease incidence was low and showed an increase in frequency by year, which we attributed to more observing and complete recording by the owner, rather than an actual increase in disease incidence. Most diseases occurred early in lactation and their frequency increased with lactation number; the exception was udder edema, which occurred mainly during the first 2 lactations. An informal path model of disease interrelationships was made conditional on herd. Based on the results we inferred 2 independent pathways: one started by udder edema, and the other by milk fever. Udder edema was directly associated with mastitis occurrence from 0 to 30 d in lactation, metritis, and cystic ovaries. Mastitis from 0-30 d in lactation increased the risk of both mastitis from 31-150 d in lactation and cystic ovaries. Both of these increased the risk of late lactation mastitis. Milk fever was directly related with displaced abomasum, which increased the risk of footrot. In general, diseases that occurred in early lactation tended to increase the risk of other diseases later in lactation. PMID- 10480461 TI - Pharmacokinetics of enrofloxacin given by the oral, intravenous and intramuscular routes in broiler chickens. AB - Enrofloxacin was given to broiler chickens, 3 groups of 6 birds each, at a dose of 5 mg/kg. Routes of administration were intravenous (i.v.), intramuscular (i.m.) and oral (p.o.) and blood samples were collected from the jugular vein for determination of serum drug levels over a 54-hour period after administration. Drug levels were determined using Bacillus subtilis spore suspension on Meuller Hinton antibiotic medium. Intravenous administration produced drug levels which followed a bi-exponential decay according to the model C = 101e(-1.84(t)) + 1.30e(-0.06(t)). After i.m. administration, the mean Cmax observed (2.01 microg/mL) occurred at 1 h and levels were detected for up to 48 h. The mean time to maximum concentration (Tmax) for the birds occurred at 0.79 h. The model describing serum concentrations after i.m. administration was C = 1.35e(-0.48(t)) + 1.27e(-0.07(t)) - 2.06e(-2.1(t)). Serum concentrations after oral administration were lower and the mean +/- standard error of mean, of the maximum concentrations (Cmax) was 0.99 microg/mL at 2 h after administration. The mean residence times after the 3 routes of administration were not significantly different and ranged from 12.5-13.7 h. Bioavailability by the oral route was 80.1%. Dialysis of chicken plasma vs saline indicated that the protein binding was 22.7%. PMID- 10480462 TI - The use of chance-corrected agreement to diagnose canine compulsive disorder: an approach to behavioral diagnosis in the absence of a 'gold standard'. AB - This study assessed the diagnostic accuracy of formal diagnostic criteria for canine compulsive disorder (canine CD). Canine CD is a syndrome of abnormal behaviors that are believed to result from conflict or frustration. Differential diagnoses include normal conflict behavior and learned behavior. In studies of canine CD, confidence in the diagnosis comes with knowing the accuracy of the diagnostic method. This accuracy may be quantified as the chance-corrected agreement between the diagnostic method and a 'gold standard' diagnostic test. The present study examined the agreement between diagnoses of canine CD made by an expert (the 'gold standard') and by using formal diagnostic criteria. The owners of 84 dogs suspected of having CD received 2 telephone interviews. The first utilized a detailed, pre-tested questionnaire; a dog was then diagnosed with CD if the behavioral history met 7 diagnostic criteria. The second interview was given by a behavioral expert whose diagnosis was based on personal experience. The interviewers were blind to each other's diagnoses. The chance corrected agreement between diagnoses was minimal (kappa = 0.02) and disagreement was associated with 3 of the formal criteria: a history of conflict or frustration, an increase in the number of contexts that elicit the behavior, and an increase in the daily frequency of the behavior. Reasons for the disagreement include the order of the interviews, response biases, the setting of the interviews, and, possibly, the diversity of the behaviors associated with canine CD. To the authors' knowledge, this type of study is the first in clinical ethology to address validation of the diagnostic method. The results indicate 3 developmental aspects of canine CD that should be examined in future work. PMID- 10480463 TI - Lack of evidence of conserved lentiviral sequences in pigs with post weaning multisystemic wasting syndrome. AB - In order to investigate the role of retroviruses in the recently described porcine postweaning multisystemic wasting syndrome (PMWS) serum and leukocytes were screened for reverse transcriptase (RT) activity, and tissues were examined for the presence of conserved lentiviral sequences using degenerate primers in a polymerase chain reaction (PCR). Serum and stimulated leukocytes from the blood and lymph nodes from pigs with PMWS, as well as from control pigs had RT activity that was detected by the sensitive Amp-RT assay. A 257-bp fragment was amplified from DNA from the blood and bone marrow of pigs with PMWS. This fragment was identical in size to conserved lentiviral sequences that were amplified from plasmids containing DNA from several lentiviruses. Cloning and sequencing of the fragment from affected pigs, however, did not reveal homology with the recognized lentiviruses. Together the results of these analyses suggest that the RT activity present in tissues from control and affected pigs is the result of endogenous retrovirus expression, and that a lentivirus is not a primary pathogen in PMWS. PMID- 10480464 TI - Isolation of caprine arthritis encephalitis virus from goats in Mexico. AB - A lentivirus was isolated from 2 goats in Mexico that were seropositive to caprine arthritis encephalitis virus (CAEV) by the agar gel immunodiffusion (AGID) test. The lentivirus was identified as CAEV by the observation of giant multinucleated cells (syncytia) in goat synovial membrane (GSM) monolayers co cultivated with blood mononuclear (BMN) cells from the seropositive goats, and by amplifying a DNA segment of the CAEV gag gene using the polymerase chain reaction (PCR) technique. Subsequently, cell supernatants from the GSM cells co-cultivated with BMN cells were used to infect 2 CAEV-seronegative goats. These goats seroconverted to CAEV as determined by the AGID test, and CAEV was re-isolated from these goats. One of the goats developed polyarthritis 8 mo after inoculation. Previous serological surveys indicate that infection with CAEV is prevalent among goats in Mexico. To our knowledge this is the first report of CAEV isolation in Mexico. Because of globalization of markets and increased trading among nations, the rapid identification and reporting of diseases such as CAEV are important to prevent the dissemination of these diseases. PMID- 10480465 TI - Evaluation of the effect of storage at -70 degrees C for six months on hemostatic function testing in dogs. AB - Freezing is a routine method of storage for plasma that is to be used in evaluating certain aspects of hemostatic function in many species. The purpose of this study was to evaluate the effect of storage at -70 degrees C for 6 mo on canine plasma samples. On fresh and frozen plasma from 12 clinically healthy dogs, prothrombin time, activated partial thromboplastin time, thrombin clotting time, fibrinogen determination, antithrombin III activity, fragment D and E assay, and protamine sulfate test were performed. Clinical agreement analysis was utilized to determine the effect of such storage on all assays. Individual differences detected between fresh and frozen samples were all within 2 standard deviations of the mean difference. With the exception of the activated partial thromboplastin time, storing canine plasma at -70 degrees C for 6 mo has no significant effect on hemostatic function, as assessed by these tests. PMID- 10480466 TI - Low dietary magnesium is associated with insulin resistance in a sample of young, nondiabetic Black Americans. AB - In both humans and experimental animals, dietary induced magnesium deficiency is correlated with insulin resistance. The purpose of this study was to determine whether dietary magnesium intake is associated with insulin sensitivity or blood pressure in a sample of nondiabetic, young adult black Americans. We also examined dietary calcium, potassium, and sodium intake. The study was conducted on a sample (n = 179) of young adults aged 30 +/- 3.4 years who had been followed longitudinally. Nutrient intake was assessed by obtaining a 24-h recall interview of dietary intake. Intake data were entered in a nutrient analysis program (Nutritionist III), which quantitated micronutrients, macronutrients, and minerals. We classified the sample into insulin-sensitive (IS) and insulin resistant (IR) groups, according to insulin-stimulated glucose use (M) measured during insulin clamp. M correlated positively with magnesium intake in mg/kg of fat-free mass (r = 0.15, P < .05 overall; in men, r = 0.25, P < .02). There was a significant negative correlation of total dietary magnesium intake with the sum of insulin levels measured during an oral glucose tolerance test (OGTT) (r = 0.13, P < .05). When corrected for body fat, in men there was also a significant correlation of dietary magnesium intake, measured in mg/kg of fat-free mass, with the sum of insulin concentrations on the OGTT (r = -0.22, P < .05). When cases were categorically classified as IS versus IR, magnesium intake in mg/kg of fat free mass was lower in IR (2.97 +/- 1.4) than in IS (3.68 +/- 2.2; P = .022). These results suggest a possible role for dietary magnesium in insulin resistance. PMID- 10480467 TI - Magnesium supplementation attenuates, but does not prevent, development of hypertension in spontaneously hypertensive rats. AB - This study evaluated the effects of magnesium supplementation on the development of hypertension in spontaneously hypertensive rats (SHR) and assesses the actions of magnesium on extracellular and intracellular Ca2+, Na+, and K+ status. Ten week-old SHR (n = 72) and Wistar-Kyoto rats (WKY) (n = 60) were divided into four groups: WKY, Mg-WKY (WKY receiving 650 mg/L Mg(2+)supplementation), SHR, and Mg SHR (SHR receiving Mg2+). Rats were studied for 17 weeks. Serum and erythrocyte Mg2+ and Ca2+ (measured by atomic absorption spectroscopy) and Na+ and K+ (measured by flame photometry) were determined every 3 or 4 weeks. From 13 weeks of age, blood pressure (BP) was significantly elevated in SHR compared with age matched WKY. BP was reduced (P < .01) in SHR after 10 weeks of Mg2+ supplementation and at 27 weeks of age, BP in SHR was 218 (12 mm Hg v 184 +/- 9 mm Hg) in Mg-SHR. From 18 weeks of age, serum and intracellular Mg2+ levels were significantly lower (P < .05) and from 21 weeks of age, erythrocyte Ca2+ was significantly higher in SHR than in WKY. Mg2+ supplementation normalized intracellular Mg2+ and Ca2+ concentrations in SHR. BP was inversely correlated with erythrocyte Mg2+ (r = -0.74, P < .01) and positively correlated with erythrocyte Ca2+ (r = 0.78, P < .001). In conclusion, long-term Mg2+ supplementation significantly attenuates, but does not prevent, the development of hypertension in SHR. Furthermore, intracellular Mg2+ deficiency and Ca2+ overload in SHR are normalized by Mg2+ treatment. PMID- 10480468 TI - Homocysteine levels in hypertensive patients with a history of cardiac or cerebral atherothrombotic events. AB - Hypertension is one of the most important risk factors for cardiovascular morbidity and mortality. Recently it has been suggested that the amino acid homocysteine contributes to this process. This study evaluates whether elevated plasma levels of homocysteine in hypertensive patients are associated with increased risk for cardiovascular events. Fifty hypertensive patients with a documented history of cerebral or cardiac events were age and gender matched to 50 hypertensive patients with no evidence of any cerebral or cardiac event. Demographic details, duration of hypertension, presence of other risk factors, and use of antihypertensive medications were recorded for each patient. Plasma levels of homocysteine were measured by high-performance liquid chromatography technology. The two groups had similar demographic parameters, with a mean age of 64.6 +/- 9.4 years. Patients with cardiovascular events were more likely to be past smokers and to have been treated with calcium antagonists, aspirin, and nitrates. Homocysteine levels were 12.1 +/- 5.8 micromol/L in those with documented cardiovascular disease and 11.1 +/- 4.7 micromol/L in those without (P = NS). Levels of plasma homocysteine were higher in those with hypercholesterolemia (P = .03) and in smokers, and tended to be lower in those who used beta-blockers, angiotensin converting enzyme (ACE) inhibitors, diuretics, and nitrates. Thus, hyperhomocysteinemia is not a feature of hypertensive patients with atherothrombotic events and there is no support for additive or synergistic effects between these two independent risk factors. PMID- 10480469 TI - The relationship of serum androgens and ovulatory status to blood pressure in reproductive-age women. AB - The objective of this cross-sectional study was to determine the relationship between hyperandrogenemia and blood pressure in women with menstrual irregularities seen at an endocrinology clinic. Women with serum testosterone levels (T) > or = 30 ng/dL were more likely to have general obesity (odds ratio [OR] = 6.8, 95% confidence intervals [CI] = 2.2-27.2) and central obesity (OR = 14.5, 95% CI = 6.1-38.7) than euandrogenemic women. Hyperandrogenemic women (HA) had an OR of 2.4 (95% CI = 1.0-6.2) for elevated SBP and an OR of 2.7 (95% CI = 0.8-8.8) for elevated DBP, independent of age and ovulatory status. Obese HA had an OR of 4.7 (95% CI = 2.3-10.4) for elevated SBP and an OR of 2.9 (95% CI = 1.9 9.9) for elevated DBP. In conclusion, T is associated with an increased risk for obesity and central adiposity. T predicts BP elevation independent of age and ovulatory status. There was a synergistic relationship between obesity and androgens in their affect on BP. PMID- 10480470 TI - Reduction of risk of pneumonia associated with use of angiotensin I converting enzyme inhibitors in elderly inpatients. AB - Pneumonia is a major direct cause of death in the elderly. Although aspiration based on a reduced cough reflex is one of the causes of pneumonia in the elderly, there are few studies of angiotensin-I converting enzyme inhibitors (ACE inhibitors), which are antihypertensive drugs that induce cough, as a factor influencing the incidence of pneumonia in institutionalized elderly subjects. To assess the effect of ACE inhibitors and dihydropiridine calcium-channel blockers on the incidence of pneumonia, we conducted a hospital-based case-control study. Cases were 55 pneumonia patients aged > or = 65 years during a 1-year period. The controls were elderly subjects, frequency matched to the cases by age and gender (n = 220). Data were collected on known risk factors and on medication for hypertension, consisting of ACE inhibitors, calcium-channel blockers, and nonantihypertensive medication. The significance of differences in risk factors was analyzed using univariate and multivariate comparisons of cases and controls. After adjustment for potential confounding factors, the relative risk estimates for pneumonia were 0.38 (95% confidence interval [CI], 0.15-0.97) and 1.84 (95% CI, 0.89-3.78) for ACE inhibitors and calcium-channel blockers, respectively, relative to nonantihypertensive medication. The preventive effect of ACE inhibitors on pneumonia was apparent in long-acting ACE inhibitor users (0.24; 95% CI, 0.07-0.88). We conclude that ACE inhibitor use is an independent factor reducing risk of pneumonia among elderly inpatients. PMID- 10480471 TI - Effect of aspirin on blood pressure in hypertensive patients taking enalapril or losartan. AB - The ability of angiotensin converting enzyme (ACE) inhibitors to lower blood pressure may in part be due to the formation of vasodilatory prostaglandins. Inhibition of prostaglandin synthesis with aspirin may therefore theoretically attenuate the antihypertensive effect of ACE inhibitors. This trial studied the interaction between aspirin (ASA) and enalapril, an ACE inhibitor, and ASA and losartan, an angiotensin subtype 1 receptor antagonist. Seventeen essential hypertensive patients were studied, maintained on a stable dose of either enalapril (n = 7) or losartan (n = 10) monotherapy for > or =12 weeks before and throughout the study. Each patient received a 2-week course of placebo, 81 mg/day ASA, and 325 mg/day ASA, each treatment separated by a 2-week washout period. Blood pressure (BP) and serum thromboxane B2 (TXB2) samples were obtained at the end of each treatment period. Placebo was compared with each dose of ASA for each group. In both the enalapril and losartan groups, mean, systolic, and diastolic BP were unchanged with the addition of ASA. Concentrations of TXB2 were suppressed to <10% in both groups with ASA. This study demonstrates that 81 to 325 mg/day ASA exerts no significant effect on BP in essential hypertensives taking enalapril or losartan. PMID- 10480472 TI - The Pandora Project: results of the pilot study. AB - The Pandora Project was designed to develop a computer-assisted system to improve the appropriateness and effectiveness of hypertension treatment in clinical practice and to organize a database both for epidemiologic and economic assessments. The feasibility study was conducted by five general practitioners (GP) who enrolled 244 patients over a period of 6 months. The follow-up lasted 6 months. The computer system implemented provided a linkage among GP's office, hypertension unit, Ravenna Health Service databases, and a remote station. A total of 209 patients completed the follow-up period; 56% of patients were not normotensive despite the antihypertensive treatment. The prevalence of overweight, physical inactivity, and family history of high blood pressure and hypercholesterolemia was greater than 50%. Unplanned check-ups by GP occurred 9%. Six patients were admitted to the hospital eight times; 19 patients attended the casualty department 21 times. The mean total direct cost per patient was 567,800 Italian Lire (ITL) and increased to ITL 732,000 or to ITL 825,900 when lost productivity, calculated according to two different formulas, was added. This pilot study confirms the need and feasibility of implementing the Pandora Project in general practice in Ravenna. PMID- 10480473 TI - Matrix study of irbesartan with hydrochlorothiazide in mild-to-moderate hypertension. AB - The purpose of this study was to assess the safety and antihypertensive dose response effects of irbesartan and hydrochlorothiazide (HCTZ) in patients with mild-to-moderate hypertension. After a 4- to 5-week single-blind placebo lead-in period, 683 patients with seated diastolic blood pressure (SeDBP) between 95 and 110 mm Hg were randomized to receive once-daily dosing with one of 16 different double-blind, fixed combinations of irbesartan (0, 37.5, 100, and 300 mg irbesartan) and HCTZ (0, 6.25, 12.5, and 25 mg HCTZ) for 8 weeks. The primary efficacy variable was the change from baseline in trough SeDBP after 8 weeks of therapy. Data were analyzed by response surface modeling. At Week 8, mean changes from baseline in trough SeDBP (mm Hg) ranged from -3.5 for placebo, -7.1 to -10.2 for the irbesartan monotherapy groups, -5.1 to -8.3 for the HCTZ monotherapy groups, and -8.1 to -15.0 for the combination groups. Irbesartan plus HCTZ produced additive reductions in both SeDBP and seated systolic BP, with at least one combination producing greater BP reduction than either drug alone (P < .001). All treatments were well tolerated; there were no treatment-related serious adverse events. Irbesartan tended to ameliorate the dose-related biochemical abnormalities associated with HCTZ alone. In conclusion, the combination of HCTZ in doses up to 25 mg with irbesartan, in doses up to 300 mg, is safe and produces dose-dependent reductions in BP. PMID- 10480474 TI - Differential effects of morning and evening dosing of nisoldipine ER on circadian blood pressure and heart rate. AB - The time of administration of once-daily antihypertensive agents may have a significant impact on blood pressure control during awake and sleep periods. Using 24-h ambulatory monitoring, we compared the effects of morning and evening dosing of the long-acting dihydropyridine calcium channel blocker, nisoldipine extended-release (ER), on circadian blood pressure (BP) and heart rate in patients with mild-to-moderate hypertension. After completing a 3-week placebo run-in period, 85 patients were randomized to morning versus evening nisoldipine ER treatment at a fixed 20-mg dose. Patients were treated for 4 weeks, followed by crossover to the alternate dosing regimen for 4 additional weeks. Twenty-four hour ambulatory monitoring was performed at baseline and at 4 and 8 weeks after randomization. Awake and sleep times were determined by electronic activity recorders (Actigraphy). Similar least-squares (+/-SE) mean changes from baseline in 24-h BP (systolic BP/diastolic BP: -11.9/-7.4 +/- 0.6/0.5 v -11.6/-6.5 +/- 0.6/0.5 mm Hg) and heart rate (1.0/1.7 +/- 0.4/0.4 beats/min) occurred with morning and evening administration, respectively. A significantly greater effect on awake diastolic BP (systolic BP/diastolic BP: -12.6/-8.1 +/- 0.7/0.4 v -11.3/ 6.4 +/- 0.7/0.4 mm Hg; P = .16/.01) was observed with morning dosing compared with evening dosing. In addition, small increases in sleep and early morning heart rate were seen with evening compared with morning administration of nisoldipine (sleep, 3.1 +/- 0.4 v 0.4 +/- 0.4 beats/min; P < .001; early morning, 3.5 +/- 0.7 v 0.5 +/- 0.7 beats/min; P = .002). These differential effects on awake BP and sleep heart rate were also observed in patients who had normal (dippers) and elevated (nondippers) BP values during sleep. Appropriate evaluation of the efficacy and safety of long-acting antihypertensive agents is essential when evening administration is being considered. In the present study, the timing of nisoldipine ER administration had no effect on mean changes in BP and heart rate over a 24-h period. However, nisoldipine ER had some differential effects during sleep and awake periods with morning relative to evening dosing. PMID- 10480475 TI - Susceptibility to apoptosis measured by MYC, BCL-2, and BAX expression in arterioles and capillaries of adult spontaneously hypertensive rats. AB - Hypertension results in microvascular rarefaction or disappearance of microvessels. In the present study, we investigated the pathogenic role of apoptosis in hypertension-induced rarefaction of heart arterioles and capillaries of spontaneously hypertensive rats (SHR). Experiments were performed on hearts from 6-week-old, 16-week-old, and 30-week-old SHR (n = 30 rats) (SHR6, SHR16, SHR30). We used as controls 6-week-old, 16-week-old, and 30-week-old normotensive rats (WKY) (n = 30 rats) (WKY6, WKY16, WKY30). We analyzed the expression of c myc, bcl-2, and bax and in situ end-labeling DNA fragmentation in vascular smooth muscle cells of arterioles and endothelial cells of arterioles and capillaries. Endothelial cells of capillaries and endothelial and smooth muscle cells of arterioles of hypertensive animals (SHR) express more Bax protein and Myc protein than their respective normotensive controls by margins that were statistically significant. The SHR30 group expressed the lowest levels of Bcl-2 protein by a margin that was statistically significantly different from WKY30. We did not find evidence of apoptosis in arterioles or capillaries on the basis of in situ end labeling. However, our results indicated that alterations in the expression of members of the Bcl-2 family of proteins and Myc protein occurred in smooth muscle cells and endothelial cells of arterioles and capillaries of SHR. In conclusion, although evidence of apoptosis in arterioles and capillaries was not found by in situ end-labeling, our findings suggest that in hypertension they may have a higher susceptibility to apoptosis, and therefore rarefaction may be a consequence of apoptosis. PMID- 10480476 TI - Inverse relationship between potassium intake and coronary artery disease in the cholesterol-fed rabbit. AB - We tested the hypothesis that a low level of dietary potassium intake would exacerbate the severity of vascular lesion formation in rabbit coronary arteries during high cholesterol intake. Two groups of nine rabbits were studied for 6 weeks while eating a diet containing 2% cholesterol and 0.9% sodium. The normal potassium group consumed a diet containing 1.5% potassium and the low potassium group consumed a diet containing 0.4% potassium. After 6 weeks the animals were killed, the hearts were removed, and blood samples were withdrawn from the abdominal aorta immediately before removing the heart. The hearts were sectioned and slides were prepared and fixed with hematoxylin and eosin. The numbers of normal and abnormal vessels, and those with foam cells in the subintima, were counted in selected sections. Plasma potassium concentration in the normal and low potassium intake groups averaged 4.27 +/- 0.27 mmol/L and 3.90 +/- 0.11 mmol/L, respectively. No differences between the groups were observed in plasma cholesterol or body weight gain. The percentages of abnormal arteries in the groups were 4.20 +/- 0.35 in the normal intake group and 6.36 +/- 0.50 in the low intake group, 51% greater in the normal intake group (P < .001). These results support the hypothesis that low potassium intake exacerbates the severity of subintimal lesion development in the coronary arteries. PMID- 10480477 TI - Serial changes in cardiovascular and renal function of rabbits ingesting a high fat, high-calorie diet. AB - To explore the mechanisms of obesity-induced hypertension we analyzed the sequential changes in cardiovascular and renal function in adult rabbits switched to high-fat diet (HFD) for 8 weeks. Animals were housed in metabolic cages for continuous 24-h recording of arterial pressure by telemetry and daily urine collection. High-fat diet induced a progressive increase in body weight (+47%) and a rapid rise in mean arterial pressure, heart rate, and glomerular filtration rate that stabilized, respectively, at 14%, 31% and 68% greater than control values. Time-course analysis of changes in blood pressure may reveal two components of obesity-induced hypertension, an early phase related to HFD itself and a later phase related to weight gain. PMID- 10480478 TI - Detection, treatment, and control of adult hypertension in northwest Indiana. Ispat Inland/United Steelworkers of America Health Care Network. AB - Recent data from the third National Health and Nutrition Examination Survey (NHANES-IIIB) suggest that detection and treatment of hypertension is improving, but only 27.4% of American hypertensives achieved controlled blood pressure ([BP] < 140 and < 90 mm Hg). Our objective was to assess the degree of BP control in a group of steelworkers and their families whose health care is financed by a large corporation. A random sample of 792 adults (age > 18 years, average 55 +/- 1 years, 50.4% male) was selected from a roster of patients who were known (from claims data) to have visited a physician in 1995 to 1996. Office charts were reviewed by trained nurses, who abstracted dates of 4095 visits, 3352 BP readings, and 3331 prescribed medications. Filled prescriptions were identified from 54,689 claims submitted for pharmacy services. Hypertension, defined (per NHANES) as more than one BP reading of 140/90 or higher, or taking antihypertensive medication, was found in 437 (55%). At least one antihypertensive medication was prescribed for 386 (88%) of the hypertensives; only 10 failed to have any prescription for antihypertensive medications filled. Controlled hypertension, as defined by Healthcare Employer Data Information Sheet (HEDIS) 3.0 (average BP < 140 and < 90 mm Hg in the office during a year-long period of observation), was observed in 189 patients (43% of total sample, or 50% of the 382 with at least one recorded BP measurement). These data suggest that in this population, insured by a jointly run employer-union health benefits plan, Healthy People 2000's BP goal-at least 50% of hypertensives having BP under control by the turn of the century-may be achieved ahead of schedule. PMID- 10480480 TI - alpha-Tocopherol increased nitric oxide synthase activity in blood vessels of spontaneously hypertensive rats. AB - Antioxidant protection provided by different doses of alpha-tocopherol was compared by determining nitric oxide synthase (NOS) activity in blood vessels of spontaneously hypertensive rats (SHR) treated with alpha-tocopherol. SHR were divided into four groups namely hypertensive control (C), treatment with 17 mg of alpha-tocopherol/kg diet (alpha1), 34 mg of alpha-tocopherol/kg diet (alpha2), and 170 mg of alpha-tocopherol/kg diet (alpha3). Wister Kyoto (WKY) rats were used as normal control (N). Blood pressure were recorded from the tail by physiography every other night for the duration of the study period of 3 months. At the end of the trial, animals were sacrificed. The NOS activity in blood vessels was measured by [3H]arginine radioactive assay and the nitrite concentration in plasma by spectrophotometry at wavelength 554 nm using Greiss reagent. Analysis of data was done using Student's t test and Pearson's correlation. The computer program Statistica was used for all analysis. Results of our study showed that for all the three alpha-tocopherol-treated groups, blood pressure was significantly (P < .001) reduced compared to the hypertensive control and maximum reduction of blood pressure was shown by the dosage of 34 mg of alpha-tocopherol/kg diet (C: 209.56 +/- 8.47 mm Hg; alpha2: 128.83 +/- 17.13 mm Hg). Also, NOS activity in blood vessels of SHR was significantly lower than WKY rats (N: 1.54 +/- 0.26 pmol/mg protein, C: 0.87 +/- 0.23 pmol/mg protein; P < .001). Although alpha-tocopherol in doses of alpha1, alpha2, and alpha3 increased the NOS activity in blood vessels, after treatment only that of alpha2 showed a statistical significance (P < .01). Plasma nitrite concentration was significantly reduced in SHR compared to normal WKY rats (N: 54.62 +/- 2.96 mol/mL, C: 26.24 +/- 2.14 mol/mL; P < .001) and accordingly all three groups showed significant improvement in their respective nitrite level (P < .001). For all groups, NOS activity and nitrite level showed negative correlation with blood pressure. It was significant for NOS activity in hypertensive control (r = 0.735, P = .038), alpha1 (r = -0.833, P = .001), and alpha2 (r = -0.899, P = .000) groups. For plasma nitrite, significant correlation was observed only in group alpha1 (r = -0.673, P = .016) and alpha2 (r = -0.643, P = .024). Only the alpha2 group showed significant positive correlation (r = 0.777, P = .003) between NOS activity and nitrite level. In conclusion it was found that compared to WKY rats, SHR have lower NOS activity in blood vessels, which upon treatment with antioxidant alpha-tocopherol increased the NOS activity and concomitantly reduced the blood pressure. There was correlation of lipid peroxide in blood vessels with NOS and nitric oxide, which implies that free radicals may be involved in the pathogenesis of hypertension. PMID- 10480479 TI - Modulation of renal blood flow by endogenous endothelin-1 in conscious rabbits with left ventricular dysfunction. AB - The current study addresses the functional status and role of the endothelin ET(A) receptor for renal vascular function in rabbits with and without heart failure (epinephrine-induced cardiomyopathy). Under baseline conditions, the ET(A) receptor antagonist BQ-123 did not change basal renal hemodynamics, but completely prevented endothelin-1 (ET-1)-induced renal vasoconstriction. In heart failure, in the presence of elevated plasma ET-1 concentrations (P < .05), renal vasoconstriction in response to exogenous ET-1 was intact. Unlike under baseline conditions, ET(A) receptor antagonism markedly increased renal blood flow (P <.05) and decreased renal vascular resistance (P < .05) in heart failure. The current study provides new insight into the pathophysiology of renal vasoconstriction associated with heart failure and the specific role of the renal ET(A) receptor in this pathophysiologic adaptation. PMID- 10480481 TI - Total peripheral conductance mediates antihypertensive effect of nonpeptide mixed endothelin receptor antagonist in deoxycorticosterone acetate-salt hypertensive rats. AB - The relative contribution of cardiac output (CO) and total peripheral conductance (TPC) to the changes in blood pressure (BP) evoked by an intravenous injection of bosentan, a nonpeptide mixed endothelin (ET) antagonist, were investigated in conscious unrestrained deoxycorticosterone acetate (DOCA)-salt hypertensive rats and sham-control rats. Blood pressure was recorded by radiotelemetry devices and CO by ultrasonic transit-time flow probes. Bosentan significantly reduced BP (from 141 +/- 3 to 111 +/- 3 mm Hg) and increased TPC (from 1.19 +/-0.1 to 1.72 +/- 0.2 mL/min/kg/mm Hg) in DOCA-salt hypertensive rats, but not in sham rats. An increase in CO opposed the BP-lowering effect of the antagonist. The results demonstrate that the role of ET receptors in the maintenance of the hypertensive state in the DOCA-salt model of hypertension is exerted at the level of the resistance vessels and not on factors that regulate cardiac output. PMID- 10480482 TI - Potassium intake and blood pressure. PMID- 10480483 TI - Modulation of vasoconstrictor and dilator pancreatic metabolites in streptozotocine diabetic rats: effect of bradykinin blockage and NO inhibition. AB - This study was designed to investigate the effect of HOE 140 (a bradykinin beta2 receptor antagonist) and N(w)-nitro-L-arginine methyl-ester (L-NAME, a nitric oxide synthase inhibitor) on endothelial and beta-cell function in induced streptozotocine (Stz) diabetic rats. The decrease in the insulinogenic index after Stz effect (control 286.03+/-104.12 and Stz 18.22+/-10.77, P<0.001 vs. Control) was partially prevented by L-NAME (46.54+/-10.12, P<0.001) and HOE 140 (105.12+/-23.06, P<0.001). It was observed in diabetic rats: L-NAME increased the pancreatic endothelin-1 (ET-1) production and HOE 140 did not. L-NAME and HOE 140 decreased the nitric oxide (NO) synthesis, increased prostacyclin 1-2 (PGI2), and did not modify thromboxane A-2 (TxA2). These results indicate that L-NAME and HOE 140 had a protective effect on the development of diabetes in the rat. The protective effect of L-NAME and HOE 140 on the insulinogenic index could be related to ET-1, bradykinin, PGI2, and NO. PMID- 10480484 TI - Gastric prostacyclin (PGI2) prevents stress-induced gastric mucosal injury in rats primarily by inhibiting leukocyte activation. AB - We investigated whether, in rats, gastric prostacyclin (PGI2) prevented gastric mucosal injury that was induced by water-immersion restraint stress by inhibiting leukocyte activation. Gastric levels of 6-keto-PGF1alpha, a stable metabolite of PGI2, increased transiently 30 min after stress, followed by a decrease to below the baseline 6-8 h after stress. Gastric mucosal blood flow decreased to approximately 40% of the baseline level 8 h after stress. Myeloperoxidase activity was significantly increased 8 h after stress. Treatment with indomethacin before stress inhibited the increase in 6-keto-PGF1alpha levels and markedly reduced mucosal blood flow. It also markedly increased leukocyte accumulation and mucosal lesion formation. Iloprost, a stable PGI2 analog, inhibited the indomethacin-induced decrease in mucosal blood flow, mucosal lesion exacerbation, and increase in leukocyte accumulation. Nitrogen mustard-induced leukocytopenia inhibited the indomethacin-associated lesion exacerbation and the increase in leukocyte accumulation, but not the decreases in mucosal blood flow. These observations indicate that gastric PGI2 decreases gastric mucosal lesion formation primarily by inhibiting leukocyte accumulation. PMID- 10480485 TI - Differential expression of prostaglandin receptor mRNAs during adipose cell differentiation. AB - To clarify the molecular basis for the prostaglandin (PG) mediated effects in adipose cells at various stages of their development, expression of mRNAs encoding receptors specific for prostaglandin E2, F2alpha and I2 (i.e. EP, FP, and IP receptors) was investigated in differentiating clonal Ob1771 pre adipocytes, as well as in mouse primary adipose precursor cells and mature adipocytes. We have further characterized the differential expression of mRNAs encoding three subtypes of the EP receptor, i.e. EP1, EP3, and EP4, and examined the expression of mRNAs encoding the three isoforms (alpha, beta, and gamma) of the EP3 receptor. Altogether the results show that the expression of IP, FP, EP1, and EP4 receptor mRNAs was considerably more pronounced in pre-adipose cells than in adipose cells, mRNAs encoding the alpha, beta, and gamma isoforms of the EP3 receptor were all exclusively expressed in freshly isolated mature adipocytes. These data may indicate that PGI2, PGF2alpha, and PGE2 may interact directly with specific receptors in pre-adipose cells, whose transduction mechanisms are known to affect maturation related changes. In mature adipocytes, however, the equipment of mRNAs encoding the EP3 receptor isoforms is in agreement with the well known effect of PGE2 on adenylate cyclase and lipolysis in mature adipocytes. PMID- 10480486 TI - Indirect enhancement of neutrophil activity and adhesion to cultured human umbilical vein endothelial cells by isoprostanes (iPF2alpha-III and iPE2-III). AB - Isoprostanes are metabolites of arachidonic acid found in blood under various conditions of oxidative stress. Because arachidonic acid derivatives are major mediators of inflammation, we investigated the potential inflammatory effects of iPF2alpha-III (previously 8-isoPGF2alpha) and iPE2-III (8-isoPGE2) on human polymorphonuclear granulocytes (PMN), as well as on human umbilical vein endothelial cells (HUVECs). The early activation marker CD11b on PMN and the adhesion molecules ICAM-1, E-selectin, and P-selectin on HUVECs were quantified by flow cytometry. Levels of the cytokines interleukin (IL)-6 and IL-8 were measured in the culture supernatant by enzyme-linked immunosorbent assay. Furthermore, adhesion of PMN to HUVECs was assessed. Neither isoprostane showed any direct stimulatory effects on PMN or HUVECs at concentrations of 0.1 or 1 microM: there was no acute elevation in expression of CD11b or P-selectin and no change of ICAM-1 or E-selectin after 4 or 24 h of incubation, respectively. The levels of interleukin IL-6 and IL-8 were also unaltered. However, PMN adhesion was significantly enhanced both after 4 and 24 h of incubation of HUVECs with iPF2alpha-III, and CD11b expression on PMN was elevated by contact of these cells with the supernatant of pre-exposed HUVECs. Neither of these actions were inhibited by an endothelin receptor antagonist (bosentan) or a combined thromboxane A2/isoprostane-receptor antagonist (SQ29548). Thus, although not having a direct pro-inflammatory potential, isoprostanes might indirectly accentuate PMN stimulation. This seems to occur via a receptor-independent mechanism, perhaps the production of an active metabolite of isoprostanes by endothelial cells. PMID- 10480487 TI - Effects of long-term supplementation of eicosapentanoic and docosahexanoic acid on the 2-, 3-series prostacyclin production by endothelial cells. AB - We studied the effects of polyunsaturated fatty, acids such as arachidonic acid [20:4 (n-6)], eicosapentanoic acid [EPA, 20:5 (n-3)], and docosahexanoic acid [DHA, 22:6 (n-3)] on the changes of lipid profiles and prostacyclin production by cultured bovine aortic endothelial cells. The amounts of 6-keto-prostaglandin F1alpha(6-keto-PGF1alpha) and delta17-6-keto-PGF1alpha, non-enzymatic metabolites of prostacyclin (PGI2 and PGI3) in culture medium were measured by gas chromatography/selected ion monitoring. Endothelial cells were supplemented for five passages with arachidonic acid, EPA, or DHA, and the fatty acids of cell lipids and prostacyclin production in cultured medium were quantified. From the fatty acid analysis, the amounts of docosapentaenoic acid [22:5 (n-3)] were significantly increased in EPA-grown cells. In DHA-grown cells, the amounts of EPA were slightly increased compared to control cells. These cells produced similar amounts of PGI2 as the controls, but larger amounts of PGI3 under basal conditions. These findings suggest that EPA, docosapentaenoic acid, and DHA are interconverted to each other, and anti-aggregatory effects of EPA or DHA may be partially due to the stimulation of prostacyclin formation in endothelial cells. PMID- 10480488 TI - Calcium dependency of arachidonic acid incorporation into cellular phospholipids of different cell types. AB - Ca2+ -independent phospholipase A2 (iPLA2) is involved in the incorporation of arachidonic acid (AA) into resting macrophages by the generation of the lysophospholipid acceptor. The role of iPLA2 in AA remodeling in different cells was evaluated by studying the Ca2+ dependency of AA uptake from the medium, the incorporation into cellular phospholipids, and the effect of the iPLA2 inhibitor bromoenol lactone on these events. Uptake and esterification of AA into phospholipids were not affected by Ca2+ depletion in human polymorphonuclear neutrophils and rat fibroblasts. The uptake was Ca2+ independent in chick embryo glial cells, but the incorporation into phospholipids was partially dependent on extracellular Ca2+. Both events were fully dependent on extra and intracellular Ca2+ in human platelets. In human polymorphonuclear neutrophils, the kinetics of incorporation in several isospecies of phospholipids was not affected by the absence of Ca2+ at short times (<30 min). The involvement of iPLA2 in the incorporation of AA from the medium was confirmed by the selective inhibition of this enzyme with bromoenol lactone, which reduced < or =50% of the incorporation of AA into phospholipids of human neutrophils. These data provide evidence that suggests iPLA2 plays a major role in regulating AA turnover in different cell types. PMID- 10480490 TI - Contribution of type II PLA2 to prostaglandin formation: a study using a type II PLA2 specific inhibitor SB 203347. AB - Arachidonic acid (AA) mobilization by phospholipase A2 (PLA2) and subsequent prostaglandin synthesis is considered to be a pivotal event in inflammation. The purpose of this study was to assess the efficacy of a Type II PLA2 specific inhibitor, SB 203347, in reducing prostaglandin production in Type II PLA2 transfected Chinese hamster ovary (CHO) cells and in human placenta. In both experimental models utilised, Type II PLA2 represents the principal isozyme contributing to total PLA2 enzymatic activity. PLA2 enzymatic activity released into cell culture media and placental explant media was quantified by radiolabelled substrate assay [14C-phosphatidylethanolamine (PE)]. Immunoreactive prostaglandin F2alpha (PGF2alpha) concentrations were determined by radioimmunoassay. SB 203347 (at 0.1-10 microM final concentration) inhibited PLA2 enzymatic activity released by Zn++ -activated CHO cells by up to 60% (P<0.0001). The concentration of PGF2alpha present in culture media was concomitantly reduced by up to 90% (P<0.0001). Similar results were observed for human placental explants. Treatment of human placental explants with SB 203347 (1 microM final concentration) significantly reduced PLA2 enzymatic activity recovered in media after 24 h incubation (P<0.0001; n = 10). Incubation media PGF2alpha concentrations were also reduced by 60% (P<0.00001). The addition of endogenous arachidonic acid (30 microM final concentration) significantly attenuated SB 203347-inhibition of PGF2alpha release (P<0.01). The data obtained in this study are consistent with the hypothesis that Type II PLA2 contributes to the liberation of arachidonic acid for prostanoid formation in human placenta and in cells that abundantly express this isozyme. PMID- 10480489 TI - The binding of apolipoprotein H (beta2-Glycoprotein I) to lipoproteins. AB - Beta2-glycoprotein I has a high affinity for triglyceride-rich particles, activates lipoprotein lipase, and is also defined as an apolipoprotein H. Previous studies have shown that apolipoprotein H is a regular structural component of the major classes of lipoproteins. In view of these findings, we analyzed the interactions of apolipoprotein H with lipoproteins in the fasting plasma of eight normal, seven hypertriglyceridemic, and seven hypercholesterolemic subjects. After rate-zonal, density gradient ultracentrifugation, apolipoprotein H was little distributed among the different density fractions, and most of it was recovered in the last fraction that contained the lipoprotein-free plasma. A small percentage (4-13%) of the apolipoprotein H associated with plasma lipoproteins was detected at the density ranging from 1.090 to 1.225 g/ml. This result means that apolipoprotein H is little associated with lipoproteins. PMID- 10480491 TI - TNFalpha-induced cyclooxygenase 2 not only increases the vasopermeability of blood-brain barrier but also enhances the neutrophil survival in Escherichia coli induced brain inflammation. AB - In Escherichia coli-induced brain inflammation, cyclooxygenase-2 was induced not only on brain arterioles at 3 h, but also on infiltrating neutrophils at 9 h post intracerebral injection. Intravenous injection of E. coli or recombinant TNFalpha also induced cyclooxygenase-2 expression on arterioles. Cyclooxygenase-2 and TNFalpha were co-localized on the arterioles as well as the infiltrating neutrophils by serial-section staining, indicating that cyclooxygenase-2 was induced by TNFalpha. NS398 (a cyclooxygenase-2 selective inhibitor) not only inhibited the increase of blood-brain barrier permeability, but also enhanced the apoptosis of the infiltrating neutrophils after E. coli stimulation. This suggests that TNFalpha-stimulated cyclooxygenase-2 induction play an important role on E. coli-induced brain inflammation. Its inhibition would help the resolution of neutrophil-mediated brain inflammation. PMID- 10480492 TI - Technology applications in intervention for preschool-age children with language disorders. AB - Preschool-age children are capable of using computers and benefiting from developmentally appropriate software. Computer technology has been used successfully in therapy for young children with speech and language disorders; however, the clinician is a crucial factor in such success. Clinicians choose communication goals and appropriate software, plan precomputer activities, provide appropriate models and opportunities during the computer activity, and provide postcomputer activities that ensure the generalization of new skills. In this article, we discuss and illustrate the characteristics of developmentally appropriate computer-based activities and the role of clinicians in planning and implementing these activities for young children. PMID- 10480493 TI - Use of technology to facilitate language skills in school-age children. AB - Technology offers numerous possibilities for facilitating language and literacy skills in school-age children and adolescents. Narrative skills can be addressed by the use of specialized programs as well as generic word processing software. Writing skills can be targeted due to the variety of possibilities for input as well as options for supporting spelling and syntax. Reading skills might be improved by the use of programs that focus on decoding, as well as hypertext tools that highlight the use of text patterns or signals. PMID- 10480494 TI - Use of technology in phonological intervention. AB - Microcomputers can support phonological intervention in a variety of ways. Software and hardware can assist clinicians in identifying errors and establishing appropriate treatment targets. Technology can also help to determine the nature of errors so that optimal intervention methods are used. Specific technology tools for addressing phonological errors due to problems with articulation, phonemic identification, and phonetic mapping are discussed. The use of technology to document and analyze treatment performance is also addressed. PMID- 10480495 TI - Technological applications in the treatment of acquired neurogenic communication and swallowing disorders in adults. AB - Clinicians can use the computer as an effective clinical tool by incorporating what is known about neurogenic communications disorders, treatment, and technology. Computers can be used to administer activities designed by clinicians, vary stimulus characteristics, adjust response requirements, present cues, and select tasks, all in response to patient performance. Specialized devices can be used to measure small physiologic changes, help patients communicate with and control their environment, and allow clinicians to view closely what we could only imagine only a few years ago. Users of technology must focus not only on effectiveness and operational efficiency, but also ensure an optimal quality of treatment. This article reviews many of the ways technology is used in the treatment of people suffering from neurogenic communication and swallowing problems. PMID- 10480496 TI - Clinical technologies for the reduction of stuttering and enhancement of speech fluency. AB - Among the clinical technologies for the treatment of fluency disorders presented in this article are systems for assisting and automating feedback on stuttering severity, establishing speech changes that enhance fluency, and modifying speech related sensory feedback for fluency enhancement. Some additional technologies, which have been implemented only in research but are expected to find their way to clinicians in the near future, are also discussed. The effectiveness of microcomputer applications for the treatment of stuttering has received little research; however, some answers about their effectiveness may be inferred from descriptions of their operational features and published specifications. PMID- 10480497 TI - Technology in the treatment of voice disorders. AB - Currently there are several instruments that provide technological support for the treatment of patients with voice disorders. In this article, specific disorders are discussed along with the technologies that can be utilized to augment treatment. Not all forms of voice disorder can be covered, but representative samples from the major categories should provide sufficient information to encourage generalization to types that are not discussed. PMID- 10480498 TI - Screening for asymptomatic coronary artery disease in diabetes. PMID- 10480499 TI - Silent myocardial ischemia in patients with diabetes: who to screen. AB - OBJECTIVE: Silent myocardial ischemia (SMI) is more common in diabetic patients than in the general population. However, the exact prevalence of SMI is not known, and routine screening is costly. The purpose of this 1-year study was to estimate the prevalence of SMI and define a high-risk diabetic population by systematically testing patients with no symptoms of coronary artery disease (CAD). RESEARCH DESIGN AND METHODS: The criteria for inclusion in this study were age (between 25 and 75 years), duration of diabetes (>15 years for type 1 diabetes, 10 years for type 2 diabetes with no cardiovascular risk factors, and 5 years for type 2 diabetes with at least one cardiovascular risk factor), and absence of clinical or electrocardiogram (ECG) symptoms of CAD. For 1 year, 203 patients were screened, including 28 women and 45 men with type 1 diabetes (aged 41.5+/-10.9 years, mean duration of diabetes 20.9+/-7.7 years [mean +/- SD]) and 61 women and 69 men with type 2 diabetes (aged 60.7+/-8.7 years, duration of diabetes 16.5+/-7.1 years). Exercise ECG was the first choice for screening method. If exercise ECG was not possible or inconclusive, thallium myocardial scintigraphy (TMS) with exercise testing and/or dipyridamole injection was performed. If any one of these tests was positive, coronary angiography was carried out and was considered to be positive with a stenosis of > or =50%. RESULTS: Positive screening results were obtained in 32 patients (15.7%). Coronary angiography demonstrated significant lesions in 19 patients (9.3%) and nonsignificant lesions in 7 patients (1 false-positive result for exercise ECG and 6 false-positive results for TMS). Coronary angiography was not performed in six patients. All but 3 of the 19 patients (15 men and 4 women) in whom silent coronary lesions were detected presented with type 2 diabetes. The main differences between the 16 type 2 diabetic patients presenting with coronary lesions and the type 2 diabetic patients without SMI were a higher prevalence of peripheral macroangiopathy (56.2 vs. 15.1%, respectively, P < 0.01) and a higher prevalence of retinopathy (P < 0.05). No correlation was found between SMI and duration of diabetes, HbA1c level, renal status, or cardiovascular risk factors except for family history of CAD. CONCLUSIONS: The results of this study allowed us to determine a high-risk group for SMI in the diabetic population. SMI with significant lesions occurs in 20.9% of type 2 diabetic male patients who are totally asymptomatic for CAD. Based on these findings, we recommend routine screening for male patients in whom the duration of type 2 diabetes is >10 years or even less when more than one cardiovascular risk factor is present. PMID- 10480500 TI - Effects of postmenopausal hormone replacement therapy on central abdominal fat, glycemic control, lipid metabolism, and vascular factors in type 2 diabetes: a prospective study. AB - OBJECTIVE: To examine the effects of hormone replacement therapy (HRT) on lipid metabolism, glycemic control, total body and central abdominal fat, blood pressure (BP), and arterial pulse wave velocity (APWV) in overweight postmenopausal females with type 2 diabetes. RESEARCH DESIGN AND METHODS: This was a 12-month prospective study of 14 subjects (mean +/- SD age 57.5+/-5.6 years, BMI 29.5+/-4.8 kg/m2) randomized to 6 months of observation or HRT before crossover. HRT consisted of 2 months of conjugated equine estrogen (CEE) 0.625 mg daily, followed by 4 months CEE and medroxyprogesterone 5 mg daily. Measures included anthropometry, fasting glucose, insulin, HbA1c, total and HDL cholesterol, triglycerides, apolipoprotein B, LDL particle size, nonesterified fatty acids (NEFA), sex hormone-binding globulin, resting energy expenditure (REE), total and central abdominal fat (by dual-energy X-ray absorptiometry), resting BP, APWV (by applanation tonometry), physical activity, well-being, and sexual function. RESULTS: Six months of HRT resulted in significant reductions in waist-to-hip ratio (-0.03+/-0.01 vs. 0.01+/-0.009, P = 0.007), HbA1c (-0.34+/ 0.24 vs. 0.6+/-0.4%, P = 0.04), total cholesterol (-0.6+/-0.1 vs. 0.2+/-0.2 mmol/l, P = 0.001), central abdominal fat (-175+/-51 vs. -24+/-56 g, P = 0.05), and improved physical functioning (P = 0.05), compared with observation. There was a minor increase in REE with HRT (33+/-23 vs. -38+/-23 kJ/day, P = 0.04). Total fat mass, fasting glucose, insulin, triglyceride, apolipoprotein B, NEFA, resting BP, APWV, and physical activity were unchanged. CONCLUSIONS: Postmenopausal HRT in these overweight women with type 2 diabetes was associated with a reduction in central adiposity and improvement in lipid metabolism and glycemic control without deterioration in weight status or cardiovascular parameters measured. Whether HRT-induced improvements in these cardiovascular risk factors result in lower long-term cardiovascular morbidity and mortality, as observed in nondiabetic women, awaits further study. PMID- 10480501 TI - Perioperative glycemic control and the risk of infectious complications in a cohort of adults with diabetes. AB - OBJECTIVE: Although hyperglycemia is hypothesized to increase the short-term risk of infection, this hypothesis has not been well tested in a clinical setting. This study was designed to assess the relationship of perioperative glycemic control to the subsequent risk of infectious complications. RESEARCH DESIGN AND METHODS: A total of 411 adults with diabetes who underwent coronary artery surgery from 1990 to 1995 in the cardiac surgery service of an urban university hospital were included in a nonconcurrent prospective cohort study based on chart review. Perioperative glycemic control was characterized by the mean of six capillary glucose measurements taken during the 36-h interval following surgery. The major outcomes studied were infections of leg and chest wounds, pneumonia, and urinary tract infections. RESULTS: Mean postoperative glucose levels ranged from 121 to 352 mg/dl and were divided into quartiles: quartile 1 (121-206 mg/dl), quartile 2 (207-229 mg/dl), quartile 3 (230-252 mg/dl), and quartile 4 (253352 mg/dl). After simultaneous adjustment for age, sex, race, underlying comorbidity, acute severity of illness, and the length of the stay in the surgical intensive care unit, patients with higher mean capillary glucose readings were at increased risk of developing infections. Compared with people in the lowest quartile of postoperative glucose, those in quartiles 2 (relative odds of infection [95% CI] = 1.17 [0.57-2.40]), 3 (1.86 [0.94-3.68]), and 4 (1.78 [0.86-3.47]) were at progressively higher risk for infection (P = 0.05 for trend). CONCLUSIONS: In patients with diabetes who undergo coronary artery surgery, postoperative hyperglycemia is an independent predictor of short-term infectious complications. Physicians should consider a glucose concentration target of < or =200 mg/dl to reduce the risk of infection. PMID- 10480503 TI - Metabolic and immunologic effects of insulin lispro in gestational diabetes. AB - OBJECTIVE: To compare the immunologic response to insulin lispro with that to regular human insulin, thereby assuring its safety for use in women with gestational diabetes, and to verify that it is effective. RESEARCH DESIGN AND METHODS: We compared the metabolic and immunologic effects of insulin lispro and regular human insulin in 42 women >18 years of age diagnosed with gestational diabetes by oral glucose tolerance testing at 14-32 weeks of gestation. Patients were randomized to receive regular human insulin or insulin lispro before consuming a test meal. Serum insulin, blood glucose, and C-peptide concentrations were measured. Throughout the remainder of gestation, patients received premeal insulin lispro or regular human insulin combined with basal insulin and performed blood glucose self-monitoring before and after each meal. Insulin antibodies and HbA1c were determined at enrollment and 6 weeks later. In addition, 10 patients received continuous intravenous insulin (4 lispro, 6 regular human insulin) and dextrose infusions intrapartum to assess placental insulin transfer. RESULTS: Anti-insulin antibody levels were similar in the two groups. Insulin lispro was not detectable in the cord blood. During a meal test, areas under the curve for glucose, insulin, and C-peptide were significantly lower in the lispro group. Mean fasting and postprandial glucose concentrations and end point HbA1c were similar in the two groups. The lispro group demonstrated fewer hypoglycemic episodes (symptoms and blood glucose concentrations <55 mg/dl). No fetal or neonatal abnormalities were noted in either treatment group. CONCLUSIONS: Insulin lispro may be considered a treatment option for women with gestational diabetes. PMID- 10480502 TI - Diabetes in urban African-Americans. XVII. Availability of rapid HbA1c measurements enhances clinical decision-making. AB - OBJECTIVE: To assess the impact of rapid-turnaround HbA1c results on providers' clinical decision-making and on follow-up HbA1c levels. RESEARCH DESIGN AND METHODS: The research design was a randomized clinical trial in which rapid HbA1c results were made available to providers on even days of the month (rapid, n = 575), but delayed by 24 h on odd days (conventional, n = 563). Adjustment of therapy for patients with type 2 diabetes was considered appropriate if therapy was intensified for HbA1c values >7% or not intensified for HbA1c values < or =7%. A post-hoc analysis was also performed using patients (n = 574) who returned for follow-up 2-7 months later to ascertain the effect of rapid HbA1c availability on subsequent glycemic control. RESULTS: Rapid HbA1c availability resulted in more appropriate management compared with conventional HbA1c availability (79 vs. 71%, P = 0.003). This difference was due mainly to less frequent intensification when HbA1c levels were < or =7% (10 vs. 22%, P < 0.0001) and slightly to more frequent intensification for patients with HbA1c values >7% (67 vs. 63%, P = 0.33). For both groups, intensification was greatest for patients on insulin (51%) compared with patients on oral agents (35%) and diet alone (14%) (P < 0.0001). Regression analysis confirmed that providers receiving conventional HbA1c results were more likely to intensify therapy in patients who already had HbA1c levels < or =7%. Over 2-7 months of follow-up, HbA1c rose more in patients with conventional HbA1c results compared with rapid results (0.8 vs. 0.4%, P = 0.02). In patients with initial HbA1c >7%, rapid HbA1c results had a favorable impact on follow-up HbA1c independent of the decision to intensify therapy (P = 0.03). CONCLUSIONS: Availability of rapid HbA1c determinations appears to facilitate diabetes management. The more favorable follow-up HbA1c profile in the rapid HbA1c group occurs independently of the decision to intensify therapy, suggesting the involvement of other factors such as enhanced provider and/or patient motivation. PMID- 10480504 TI - Benefits of a multidisciplinary approach in the management of recurrent diabetic foot ulceration in Lithuania: a prospective study. AB - OBJECTIVE: To assess the ability of a multidisciplinary approach to diabetic foot care to reduce the incidence of recurrent ulceration and amputations compared with standard care in a 2-year prospective study. RESEARCH DESIGN AND METHODS: A total of 145 patients with a past history of neuropathic foot ulcers but no evidence of peripheral vascular disease entered the study. Subjects were screened for their neuropathic and vascular status at baseline, and all received identical foot care education. The intervention group (n = 56) was followed by the multidisciplinary team of physicians, nurses, and podiatrists with regular podiatry and reeducation every 3 months and the provision of specialty footwear as required. The standard treatment group was followed in local clinics on a trimonthly basis and received identical screening and education at baseline. RESULTS: There were no significant differences at baseline in age (intervention 59.2+/-13.4, standard treatment 58.5+/-11.5 years), duration of diabetes (14.0+/ 7.1 vs. 15.6+/-7.8 years), or neuropathic status (vibration perception threshold [VPT]: 31.1+/-12.1 vs. 33.9+/-11.3 V, neuropathy disability score [NDS]: 8.1+/ 1.4 vs. 7.9+/-1.7). All patients had an ankle brachial pressure index (ABPI) of >0.9 and at least one palpable foot pulse. Significantly fewer recurrent ulcers were seen in the intervention group than in the standard treatment group during the 2-year period (30.4 vs. 58.4%, P < 0.001). CONCLUSIONS: This prospective study has demonstrated the effectiveness of a multidisciplinary approach to diabetic foot care together with the provision of specialty footwear in the long term management of high-risk patients with a history of neuropathic foot ulcers. PMID- 10480505 TI - Daily alcohol consumption and the risk of type 2 diabetes in Japanese men: the Osaka Health Survey. AB - OBJECTIVE: To investigate the relationship between daily alcohol consumption and the risk of type 2 diabetes in a large Japanese cohort. RESEARCH DESIGN AND METHODS: We enrolled 6,362 Japanese men aged 35-61 years who did not have diabetes, impaired fasting glucose, hypertension, or liver cirrhosis at study entry. Type 2 diabetes was defined as a fasting plasma glucose (FPG) level > or =126 mg/dl or was diagnosed by a physician. Data on alcohol consumption were obtained from questionnaires. We confirmed 456 cases of type 2 diabetes during the 62,016 person-years of follow-up. RESULTS: The relationship between daily alcohol consumption and the risk of type 2 diabetes among lean men and among men with a higher BMI was paradoxical. Among lean men (BMI < or =22.0 kg/m2), heavy drinking was associated with an increased risk of type 2 diabetes. Men who consumed > or =50.1 ml/day of alcohol had a relative risk (RR) of 2.48 (95% CI 1.31-4.71) compared with nondrinkers after adjusting for age, BMI, regular physical exercise, parental history of diabetes, smoking habits, and FPG level. However, among men with a BMI > or =22.1 kg/m2, moderate drinking (29.1-50.0 ml/day) was associated with a decreased risk of type 2 diabetes. Daily moderate drinkers had a multiple adjusted RR of 0.58 (0.39-0.87) compared with nondrinkers. CONCLUSIONS: Among men with a BMI > or =22.1 kg/m2, moderate alcohol consumption was associated with a reduced risk of type 2 diabetes, but among lean men (BMI < or =22.0 kg/m2), heavy alcohol consumption was associated with an increased risk of type 2 diabetes. PMID- 10480506 TI - Predictors of change in the neuropsychological profiles of children with type 1 diabetes 2 years after disease onset. AB - OBJECTIVE: To identify type 1 diabetes-related predictors of change in the neuropsychological profiles of children over the first 2 years of the illness. RESEARCH DESIGN AND METHODS: Children (n = 116) aged 3-14 years were assessed soon after diagnosis and re-evaluated 2 years later to examine relationships between illness variables, such as age of onset and metabolic control history, and changes in neuropsychological status over the first 2 years of type 1 diabetes. RESULTS: Illness variables were significant predictors of change in neuropsychological test scores within 2 years of onset of type 1 diabetes. Age of onset of type 1 diabetes predicted negative change on Performance Intelligence Quotient, whereas both recurrent severe hypoglycemia and chronic hyperglycemia were associated with reduced memory and learning capacity. CONCLUSIONS: These results suggest that the relationship between metabolic control and neuropsychological risk is nonlinear in that children with either recurrent severe hypoglycemia or chronically elevated blood sugars exhibit negative changes in their neuropsychological profiles. Onset of type 1 diabetes very early in life adds another dimension of risk, particularly affecting the acquisition of visuospatial skills. PMID- 10480507 TI - Discordance between physician and adolescent assessments of adherence to treatment: influence of HbA1c level. The PEDIAB Collaborative Group. AB - OBJECTIVE: To compare the subjective assessments (perceptions) of physicians and adolescent diabetic patients on the adolescents' adherence to treatment and to test the hypothesis that the HbA1c level influences physicians' perceptions. RESEARCH DESIGN AND METHODS: In a multicenter cross-sectional survey, 143 adolescents with diabetes (mean age 14.6 years) auto-assessed, while their pediatricians independently assessed, the level of adherence to treatment on a four-point scale. Scores of adherence given by a validated scale and metabolic control (HbA1c) were compared according to those assessments. RESULTS: Agreement between the adherence perceptions from adolescents and physicians was low (kappa = 0.23), and adolescents scored significantly higher (P < 0.001). Mean adherence score to diabetes treatment was significantly higher when the adolescents' perception of their self-care behaviors was good than when it was poor (P = 0.01), but did not significantly differ according to physicians' perception. Mean HbA1c level was significantly lower when the self-care behavior perception was good than when it was poor, both for the adolescents (P = 0.02) and for the physicians (P < 0.001). Multivariate analyses showed that only the adherence scale score was significantly associated with the adolescents' perception (P = 0.015), whereas only HbA1c level was significantly associated with the physicians' perception (P = 0.0008). CONCLUSIONS: By identifying the possible discrepancy between their own assessment of adherence and that of adolescents, and by avoiding the systematic attribution of poor metabolic control to poor adherence, physicians could generate a more confident and collaborative relationship with diabetic adolescents and therefore facilitate adolescents' self management. PMID- 10480508 TI - Plasma insulin, growth hormone, cortisol, and central obesity among young Chinese type 2 diabetic patients. AB - OBJECTIVE: To examine the relationships between central obesity, insulin resistance index, plasma insulin, growth hormone (GH), and cortisol concentrations in 90 young Chinese type 2 diabetic patients (aged 33+/-5 years) and 104 age- and sex-matched control subjects (aged 32+/-9 years). RESEARCH DESIGN AND METHODS: Young Chinese diabetic patients (aged <40 years) were recruited from the Prince of Wales Hospital. Blood pressure, height, weight, and waist and hip circumferences were determined. Venous blood was sampled for measurements of fasting plasma glucose, HbA1c, lipids, creatinine, insulin, GH, and cortisol. A 24-h urine was assayed for urinary albumin excretion (UAE). General and central obesity was represented by BMI and waist circumference, respectively. Insulin resistance index was estimated as a product of fasting plasma insulin and glucose concentrations. RESULTS: Compared with control subjects, diabetic patients were more obese, hyperglycemic, and had worse lipid profile, higher blood pressures, UAE, insulin resistance index, plasma insulin, and cortisol concentrations (all P < 0.001) but lower GH concentrations (P < 0.05). When analyzed as a whole group (n = 194), increasing quartiles of waist circumference were associated with increasing trends of insulin resistance index, plasma insulin, and cortisol concentrations (all P < 0.01) but a decreasing trend of plasma GH concentration (P < 0.05). Using stepwise multiple regression analysis, waist circumference was only associated with sex variable (being higher in men) in the control subjects. In the diabetic group, 51% of waist circumference was independently related to male sex and increased plasma insulin and cortisol concentrations as well as reduced plasma GH levels. CONCLUSIONS: In young Chinese type 2 diabetic patients, hyperinsulinemia, hypercortisolemia, and reduced plasma GH levels were closely associated with central obesity. Based on these findings, we postulate that maladaptive hormonal responses to rapid changes in lifestyle may have led to obesity and type 2 diabetes in these young patients. Alternatively, lifestyle-related obesity may have given rise to these hormonal changes. More studies are required to delineate the nature of these relationships. PMID- 10480509 TI - Antibodies to GAD65 and a tyrosine phosphatase-like molecule IA-2ic in Filipino type 1 diabetic patients. AB - OBJECTIVE: Type 1 diabetes is more prevalent in Europeans than it is in Asians. The disease is associated with autoantibodies to GAD65 and a protein tyrosine phosphatase-like molecule (IA-2). The frequency of GAD antibodies in Asian patients with type 1 diabetes may be lower than that in Europeans. No data are available on IA-2 antibodies in Asians. We tested antibodies to GAD65 and IA-2ic (the intracellular fragment containing the antibody epitope) in Filipino diabetic patients because this population has mixed European and aboriginal racial origins. RESEARCH DESIGN AND METHODS: A cross-sectional study of antibodies to GAD65 and IA-2ic was performed on a consecutive series of 91 type 1 diabetic patients, 74 type 2 diabetic patients, and 100 control subjects attending a diabetes clinic in Manila, the Republic of the Philippines. All subjects were <40 years of age, with a mean age +/- SD of 24.8+/-9.8, 34.3+/-5.8, and 25.8+/-8.0 years, respectively. Diagnosis of type 1 diabetes was determined clinically and confirmed by baseline C-peptide. RESULTS: Of 91 type 1 diabetic patients, antibodies to GAD65 were detected in 25 (27.4%), but antibodies to IA-2ic were found in only 8 (8.8%) (P = 0.002); neither autoantibody was detected in either the type 2 diabetic or control subjects. Of the 25 recently diagnosed type 1 diabetic patients (disease duration <2.0 years), autoantibodies to GAD65 were detected in 14 (56%), but those to IA-2ic in only 4 (16%) (P = 0.007); GAD65 antibodies were detected in only 4 (6%) of 66 patients with a longer disease duration (P = 0.0004). Comparison with recently diagnosed European type 1 diabetic patients of age and disease duration similar to that of the Filipinos indicated that IA-2ic antibodies, unlike GAD antibodies, were significantly less prevalent in Filipino type 1 diabetic patients (P = 0.0007). CONCLUSIONS: This is the first study investigating the prevalence and pattern of humoral immune response in type 1 diabetic patients from the Philippines. Antibodies to IA-2ic, unlike GAD antibodies, were infrequent. Patterns of immune responses to type 1 diabetes-associated antigens may differ worldwide, with important implications for prediction of the disease and the potential for antigen-specific therapy. PMID- 10480510 TI - Insulin sensitivity indices obtained from oral glucose tolerance testing: comparison with the euglycemic insulin clamp. AB - OBJECTIVE: Several methods have been proposed to evaluate insulin sensitivity from the data obtained from the oral glucose tolerance test (OGTT). However, the validity of these indices has not been rigorously evaluated by comparing them with the direct measurement of insulin sensitivity obtained with the euglycemic insulin clamp technique. In this study, we compare various insulin sensitivity indices derived from the OGTT with whole-body insulin sensitivity measured by the euglycemic insulin clamp technique. RESEARCH DESIGN AND METHODS: In this study, 153 subjects (66 men and 87 women, aged 18-71 years, BMI 20-65 kg/m2) with varying degrees of glucose tolerance (62 subjects with normal glucose tolerance, 31 subjects with impaired glucose tolerance, and 60 subjects with type 2 diabetes) were studied. After a 10-h overnight fast, all subjects underwent, in random order, a 75-g OGTT and a euglycemic insulin clamp, which was performed with the infusion of [3-3H]glucose. The indices of insulin sensitivity derived from OGTT data and the euglycemic insulin clamp were compared by correlation analysis. RESULTS: The mean plasma glucose concentration divided by the mean plasma insulin concentration during the OGTT displayed no correlation with the rate of whole-body glucose disposal during the euglycemic insulin clamp (r = 0.02, NS). From the OGTT, we developed an index of whole-body insulin sensitivity (10,000/square root of [fasting glucose x fasting insulin] x [mean glucose x mean insulin during OGTT]), which is highly correlated (r = 0.73, P < 0.0001) with the rate of whole-body glucose disposal during the euglycemic insulin clamp. CONCLUSIONS: Previous methods used to derive an index of insulin sensitivity from the OGTT have relied on the ratio of plasma glucose to insulin concentration during the OGTT. Our results demonstrate the limitations of such an approach. We have derived a novel estimate of insulin sensitivity that is simple to calculate and provides a reasonable approximation of whole-body insulin sensitivity from the OGTT. PMID- 10480511 TI - Age-related increase in visceral adipose tissue and body fat and the metabolic risk profile of premenopausal women. AB - OBJECTIVE: Age-related differences in body fat and, more specifically, in the accumulation of abdominal visceral adipose tissue (AT) were examined as potential covariates of the age-related difference in the metabolic profile predictive of cardiovascular disease (CVD) risk observed in young, as compared with middle aged, premenopausal women. RESEARCH DESIGN AND METHODS: Body composition, AT distribution, plasma lipoprotein-lipid levels, glucose tolerance, and plasma insulin concentrations were assessed in a sample of 122 young women (27.4+/-7.5 years, mean +/- SD) and compared with a sample of 52 middle-aged premenopausal women (49.5+/-5.3 years) who still had a normal menstrual cycle. RESULTS: Middle aged women were characterized by elevated levels of total abdominal and visceral AT and greater body fat mass and waist circumference, as well as by higher plasma levels of total cholesterol, LDL cholesterol, apolipoprotein (apo)B, and LDL-apoB compared with younger women. Furthermore, middle-aged women showed a greater glycemic response to a 75-g oral glucose load than young women (P < 0.01). In both young and middle-aged subjects, visceral AT accumulation was significantly correlated with plasma triglyceride, apoB, and LDL-apoB levels and with the cholesterol/HDL cholesterol ratio, as well as with plasma glucose, insulin, and C peptide levels measured in the fasting state and after the oral glucose load, and negatively correlated with HDL cholesterol levels (-0.41 < or = r < or = 0.65, P < 0.05). When variables were adjusted for levels of visceral AT and fat mass, age related differences that were initially found in plasma apoB and LDL-apoB levels, as well as in fasting glycemia and glucose tolerance, were eliminated. CONCLUSIONS: Results of the present study suggest that even before the onset of menopause there is an age-related deterioration in the metabolic risk profile and an increase in visceral AT deposition in middle-aged women compared with young control subjects. Furthermore, our results provide support for the notion that the age-related increase in visceral AT accumulation is a significant factor involved in the deterioration of the CVD risk profile noted in premenopausal women with age. PMID- 10480512 TI - Risk factors for severity of diabetic polyneuropathy: intensive longitudinal assessment of the Rochester Diabetic Neuropathy Study cohort. AB - OBJECTIVE: Chronic hyperglycemia relates to the occurrence of diabetic polyneuropathy (DPN), but has not yet been shown to relate to its overall severity In addition, the degree and duration of hyperglycemia, which measure of chronic hyperglycemia is most predictive of defined levels of severity of DPN, and which other putative risk factors are involved remain unknown. RESEARCH DESIGN AND METHODS: In a longitudinal study of 264 diabetic individuals in Rochester, MN, risk factors and other diabetic complications assessed at regular intervals during an average of approximately 7 years were tested for their association with a composite score of severity of DPN at the last examination. RESULTS: In multivariate analysis, diabetic retinopathy severity level (at last examination), mean ln(24-h proteinuria x duration of diabetes), and mean GHb were the main covariates for severity of DPN (R2 = 0.33). Excluding markers of microvessel and macrovessel disease, the independent risk factors were mean In(GHb x duration of diabetes), GHb, and type of diabetes (R2 = 0.23). CONCLUSIONS: We found that diabetic microvessel disease, chronic hyperglycemia exposure, and type of diabetes are associated with severity of DPN, and we believe these factors are implicated in its cause. Each of the five markers of microvessel disease was a strong covariate for severity of DPN. Mean GHb predicts severity of DPN better than duration of diabetes, and the latter predicts severity of DPN better than mean fasting plasma glucose. Knowing the severity of microvessel disease, the degree of chronic hyperglycemia exposure, and the type of diabetes provides useful information to evaluate whether a coexisting polyneuropathy and its severity is probably due to diabetes. PMID- 10480513 TI - Diabetes and disenrollment in a health maintenance organization setting: a 4-year longitudinal study with a matched cohort. AB - OBJECTIVE: The increasing enrollment of Medicare beneficiaries in health maintenance organizations (HMOs) in recent years has caused concern about whether HMOs and their providers have created an unfavorable environment for members who are chronically ill. This study was designed to examine whether there are any differences in disenrollment rates among enrollees with diabetes and enrollees without diabetes. RESEARCH DESIGN AND METHODS: This was a 4-year longitudinal follow-up study with a matched cohort. Medicare beneficiaries (aged > or =65 years) with diabetes identified through pharmacy records in 1994 were matched with a comparison group according to age, sex, comorbidities, and type of provider groups in an HMO in California. RESULTS: The overall distribution of the characteristics of members in the diabetic and matched nondiabetic group is almost identical. The matched-pair chi2 tests indicated that there were no statistical differences in disenrollment rates between diabetic and nondiabetic members during all three follow-up periods (P = 0.16-0.85). CONCLUSIONS: We found that the HMO members with diabetes did not disenroll from the HMO at a higher rate than those without diabetes. The findings should alleviate some of the concern that HMOs and their contracted providers have created an unattractive environment for members who have chronic diseases such as diabetes. PMID- 10480514 TI - Risk of diabetes in the new diagnostic category of impaired fasting glucose: a prospective analysis. AB - OBJECTIVE: To prospectively evaluate progression to diabetes in individuals with impaired glucose regulation as defined according to fasting glucose alone or an oral glucose tolerance test (OGTT) (i.e., both fasting and postload glucose) to compare the ability of these two screening methods to identify people at high risk of developing diabetes. RESEARCH DESIGN AND METHODS: A working population of 1,245 nondiabetic telephone company employees aged 40-59 years was studied by OGTT in 1980. Participants were classified according to baseline fasting glucose only (as encouraged by the American Diabetes Association [ADA]) or OGTT (as recommended by the 1998 World Health Organization [WHO] consultation). Progression to diabetes was evaluated 11.5 years later according to the 1997 ADA criteria of a fasting plasma glucose level > or =7.0 mmol/l. RESULTS: With the use of the OGTT, baseline prevalence of impaired glucose regulation was substantially higher than that with fasting glucose alone (7.2 vs. 3.2%); the two groups only overlap for 40.9% of the cases because a fairly large number of people with postload hyperglycemia (59.1%) have normal fasting glucose. Progression to diabetes in participants with normal fasting glucose and postload hyperglycemia is significantly more frequent than that of people with normoglycemia (32.5 vs. 7.2%; P < 0.001) and not significantly different from that of people with both fasting and postload hyperglycemia (i.e., 44.0%). However, the former are not identified as being at unusually high risk of diabetes unless an OGTT is performed. When the use of fasting glucose alone or OGTT was validated as a marker of progression to diabetes, sensitivity was substantially higher for the OGTT (33.3 vs. 9.0%) without major differences in specificity (92.6 vs. 97.0%). CONCLUSIONS: These data (the only data so far available in Caucasians) support the viewpoint that for the identification of people at high risk of diabetes, the use of the OGTT should be maintained. PMID- 10480515 TI - Diabetes in urban African-Americans. XVI. Overcoming clinical inertia improves glycemic control in patients with type 2 diabetes. AB - OBJECTIVE: Diabetes care can be limited by clinical inertia-failure of the provider to intensify therapy when glucose levels are high. Although disease management programs have been proposed as a means to improve diabetes care, there are few studies examining their effectiveness in patient populations that have traditionally been underserved. We examined the impact of our management program in the Grady Diabetes Unit, which provides care primarily to urban African American patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: We assessed glycemic outcomes in patients with type 2 diabetes who had an intake evaluation between 1992 and 1996 and who were identified on the basis of compliance with keeping the recommended number of return visits. For 698 patients, we analyzed changes in HbA1c values between baseline and follow-up visits at 6 and 12 months, and the proportion of patients achieving a target value of < or =7.0% at 12 months. Since a greater emphasis on therapeutic intensification began in 1995, we also compared HbA1c values and clinical management in 1995-1996 with that of 1992 1994. RESULTS: HbA1c averaged 9.3% on presentation. After 12 months of care, HbA1c values averaged 8.2, 8.4, 8.5, 7.7, and 7.3% for the 1992-1996 cohorts, respectively, and were significantly lower compared with values on presentation (P < 0.0025); the average fall in HbA1c was 1.4%. The percentage of patients achieving a target HbA1c < or =7.0% improved progressively from 1993 to 1996, with 57% of the patients attaining this goal in 1996. Mean HbA1c after 12 months was 7.6% in 1995-1996, significantly improved over the level of 8.4% in 1992-1994 (P < 0.0001). HbA1c levels after 12 months of care were lower in 1995-1996 versus 1992-1994, whether patients were managed with diet alone, oral agents, or insulin (P < 0.02). Improved HbA1c in 1995-1996 versus 1992-1994 was associated with increased use of pharmacologic therapy CONCLUSIONS: Structured programs can improve glycemic control in urban African-Americans with diabetes. Self examination of performance focused on overcoming clinical inertia is essential to progressive upgrading of care. PMID- 10480516 TI - Insulin aspart (B28 asp-insulin): a fast-acting analog of human insulin: absorption kinetics and action profile compared with regular human insulin in healthy nondiabetic subjects. AB - OBJECTIVE: To study the pharmacokinetic and pharmacodynamic profile of insulin aspart (a new fast-acting human insulin analog) after subcutaneous administration in the deltoid, abdominal, and thigh sites and to compare this profile with regular human insulin (Novolin; Novo Nordisk A/S, Copenhagen). RESEARCH DESIGN AND METHODS: A total of 20 healthy subjects were studied in a single-center six period double-blind randomized crossover trial with 6 study days and a washout period of 1 week between each single daily dose of the trial drug. Subjects were randomized to receive a single dose of 0.2 U/kg of insulin aspart or regular insulin on each of the 6 study days in three different sites (the deltoid, the abdomen, and the thigh) during a 10-h euglycemic clamp (two drugs and three injection sites). Pharmacokinetic and pharmacodynamic measurements were derived from blood sample measurements of glucose, insulin, and C-peptide during these clamps. RESULTS: The pharmacodynamic data from the euglycemic clamp study showed that, regardless of injection site, the maximal glucose infusion rate (GIR Cmax) was greater and occurred at an earlier time (GIR Tmax) after administration of insulin aspart than regular insulin (GIR Cmax: abdomen 813 vs. 708, deltoid 861 vs. 736, and thigh 857 vs. 720 g/min, P < 0.05 for all; GIR Tmax: abdomen 94 vs. 173, deltoid 111 vs. 192, and thigh 145 vs. 193 g/min, P < 0.05 for all). Pharmacokinetic parameters were also consistent with faster absorption and higher peak insulin concentrations after insulin aspart administration. From all sites, the peak insulin concentration (Cmax) was higher and occurred earlier (Tmax) after administration of insulin aspart than of regular insulin (Cmax: abdomen 501 vs. 260, deltoid 506 vs. 252, thigh 422 vs. 220 pmol/l, P < 0.001 for all sites; Tmax: abdomen 52 vs. 109, deltoid 54 vs. 98, and thigh 60 vs. 107 min, P < 0.01 for all sites). The absorption and glucose-lowering action of insulin aspart did not differ between sites (similar GIR Cmax, Tmax, and area under the curve parameters). However, the duration of the glucose-lowering effect was up to 34 min shorter (P < 0.01) for the abdomen injections than for the deltoid or thigh injections (lower time of 50% glucose disposal). In addition, the amount of glucose infused was significantly lower by 10-14% in the abdomen than in other sites. CONCLUSIONS: Subcutaneous administration of insulin aspart causes a more rapid and intense maximal effect compared with regular insulin during euglycemic clamp studies in nondiabetic subjects. Abdominal administration of insulin aspart has a shorter duration of glucose-lowering effect compared with administration in the deltoid or thigh. PMID- 10480517 TI - Usefulness of ambulatory blood pressure monitoring in pregnant women with type 1 diabetes. AB - OBJECTIVE: Pregnancy in type 1 diabetes is associated with an increased risk of developing pregnancy-induced hypertension (PIH). Ambulatory blood pressure monitoring (ABPM) has been used to screen for preeclampsia in nondiabetic pregnancy. To date, there are no data regarding ABPM during pregnancy in normotensive type 1 diabetic women. This study sought to establish blood pressure (BP) profiles for pregnant type 1 diabetic women using ABPM and determine whether the BP pattern can define a population at risk for developing PIH. RESEARCH DESIGN AND METHODS: ABPM was carried out for one 24-h period during each trimester--in the first trimester between weeks 7 and 12, in the second trimester between weeks 20 and 24, and in the third trimester between weeks 30 and 34--in 22 normotensive pregnant type 1 diabetic and 10 pregnant nondiabetic women. RESULTS: The incidence of PIH was fourfold greater in type 1 diabetic women than in control subjects. Diabetic women showed higher daily diastolic BP in the third trimester compared with nondiabetic pregnant women. Diabetic women who developed PIH in the third trimester showed significantly higher BP profiles throughout gestation than those who remained normotensive. Receiver operator characteristics curves for nighttime systolic BP showed the best predictive capacity for PIH, with a cutoff > 105 mmHg (85% sensitivity and 92% specificity). CONCLUSIONS: Our study confirms the early increase of BP in patients who will develop PIH and suggests that nighttime systolic BP >105 mmHg in the second trimester is a useful predictor of PIH. ABPM may be useful in screening for PIH in pregnant diabetic women. PMID- 10480518 TI - Glomerular filtration rate, urinary albumin excretion rate, and blood pressure changes in normoalbuminuric normotensive type 1 diabetic patients: an 8-year follow-up study. AB - OBJECTIVE: To analyze the changes in glomerular filtration rate (GFR), urinary albumin excretion rate (UAER), and blood pressure (BP) levels in a cohort of normoalbuminuric and normotensive type 1 diabetic patients. RESEARCH DESIGN AND METHODS: This is an 8.4+/-2.1-year prospective study of 33 normotensive normoalbuminuric (24-h UAER <20 microg/min) type 1 diabetic patients. UAER (radioimmunoassay), GFR (51Cr-EDTA single-injection technique), and GHb (ion exchange chromatography) were measured at baseline and at 1- to 2-year intervals. RESULTS: The GFR decreased (137.6+/-16.5 to 116.4+/-21.3 ml x min(-1) x 1.73 m( 2) P < 0.05) during the follow-up period. GFR reduction (-0.20+/-0.29 ml x min( 1) x month(-1); P < 0.05) was associated with baseline GFR and mean GHb (R2 = 0.30; beta = 0.072; F = 6.54; P = 0.004). UAER was higher at the end of the study (3.7-7.1 microg/min; P = 0.017). Microalbuminuria was observed in two patients, while macroalbuminuria was observed in one. No changes in UAER were observed when these three patients were excluded from the analysis. Mean blood pressure (MBP) increased during the study (85.8+/-9.7 to 99.6+/-11.6 mmHg; P < 0.001). MBP at the end of the study was associated with age and GFR at baseline (R2 = 0.39; beta = 0.074; F = 9.64; P = 0.001). CONCLUSIONS: In this cohort of normoalbuminuric normotensive type 1 diabetic patients, GFR decreased and BP levels increased during the follow-up period. The predictors for the GFR change were baseline GFR level and metabolic control. For end-of-study MBP, the predictor was baseline GFR level. PMID- 10480519 TI - Immunogenetic analysis suggests different pathogenesis for obese and lean African Americans with diabetic ketoacidosis. AB - OBJECTIVE: When presenting with diabetic ketoacidosis (DKA), lean and obese patients differ in their subsequent clinical course. Although lean patients tend to remain insulin dependent, most obese patients recover endogenous insulin secretion and discontinue insulin therapy. The aim of this study was to determine whether obese African-American patients with DKA could be determined to have type 1 or type 2 diabetes based on insulin secretion or the presence of immunological and genetic markers. RESEARCH DESIGN AND METHODS: This was a prospective study that analyzed the clinical characteristics, insulin secretion indices, immunological markers (islet cell, GAD, ICA512, and insulin autoantibodies), and HLA susceptibility genes (DR/DQ) in 131 patients with DKA (77 obese and 54 lean), 51 obese patients with hyperglycemia but no DKA, and 25 nondiabetic subjects. All subjects were African-American. Beta-cell function was evaluated by the C-peptide response to glucagon (1 mg i.v.) within 48 h of resolution of DKA or hyperglycemia. RESULTS: The acute C-peptide response was lower in obese DKA patients (1.0+/-0.1 ng/ml) than in obese patients with hyperglycemia (1.7+/-0.2 ng/ml, P < 0.01), but was higher than that in lean DKA patients (0.2+/-0.1 ng/ml, both P < 0.01). The overall prevalence of autoantibodies in obese subjects with DKA (17%) and obese subjects with hyperglycemia (16%) was lower than that in lean subjects with DKA (65%, P < 0.01). Obese patients with hyperglycemia and positive autoantibodies had lower rates of insulin secretion than those without antibodies. Regardless of body weight, all DKA patients with GAD autoantibodies carried the DQB1*0201 allele. However, there were no significant differences in HLA distribution between the three patient groups. CONCLUSIONS: Our results indicate that most obese African-American patients with DKA have type 2 diabetes characterized by higher insulin secretion, the absence of autoimmune markers, and a lack of HLA genetic association. In contrast, most lean African-American patients with DKA have metabolic and immunological features of type 1 diabetes. At presentation, assessment of beta-cell function and determination of autoimmune markers allow for correct classification of diabetes in African-Americans with hyperglycemic crises. PMID- 10480521 TI - ACE gene polymorphism and proliferative retinopathy in type 1 diabetes: results of a case-control study. AB - OBJECTIVE: To evaluate the relationship between the ACE insertion/deletion polymorphism and proliferative diabetic retinopathy in patients with type 1 diabetes of long duration. Based on epidemiological and pathophysiological findings, risk factors apart from glycemic control and duration of disease are likely to be involved in the development of proliferative retinopathy. RESEARCH DESIGN AND METHODS: In this case-control study, we compared 81 patients with longstanding (> or =20 years) type 1 diabetes who had nonproliferative (mild or moderate background) retinopathy with 95 patients with diabetes of similar duration and HbA1c who had proliferative retinopathy. To avoid the confounding effect of nephropathy, patients with overt nephropathy were excluded, and microalbuminuria was introduced into the multiple logistical regression model. The polymorphic region in intron 16 of the ACE gene (17q23) was analyzed using the polymerase chain reaction. RESULTS: The ACE genotype distribution in patients with proliferative retinopathy (DD 39.4%, ID 48.9%, II 11.7%) was significantly different (P < 0.001) from that of patients with nonproliferative retinopathy (DD 17.3%, ID 54.3%, II 28.4%). In a multiple logistical regression analysis, the adjusted relative risk for proliferative retinopathy in a patient with a DD genotype compared with a patient with an II genotype was 6.6 (95% CI 2.2-19.5), P = 0.0026. In addition to genotype, systolic blood pressure (odds ratio 1.027 [95% CI 1.0-1.1], P = 0.0093) but not microalbuminuria (< or =20 vs. > or =20 microg/min) reached statistical significance in the multiple regression model. Because subjects were matched regarding diabetes duration and HbA1c, we did not interpret the respective parameter estimates. CONCLUSIONS: These data provide evidence that deletion in the ACE gene is associated with the prevalence of proliferative retinopathy in type 1 diabetes and suggest that the DD genotype confers susceptibility to proliferative retinopathy independent of diabetic nephropathy PMID- 10480520 TI - Racial differences in the correlation between gonadal androgens and serum insulin levels. AB - OBJECTIVE: We previously demonstrated a direct correlation between serum insulin levels and gonadal androgens (testosterone and androstenedione) in a group of obese hyperandrogenic predominantly black women. Subsequent work by others in predominantly white women showed conflicting results. To examine these potentially important racial differences further, 14 premenopausal females from each ethnic group, of similar age, BMI, and waist-to-hip ratio, were studied. RESEARCH DESIGN AND METHODS: We measured baseline gonadal androgens, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), and leutinizing hormone (LH)/follicle-stimulating hormone ratio. Serum glucose, insulin, and C-peptide were measured at baseline and during a 2-h oral glucose tolerance test (area under the curve [AUC]). Insulin sensitivity was measured by glucose decrement during the first 15 min of an intravenous insulin tolerance test. RESULTS: Simple correlation analysis revealed a significant direct correlation in blacks (but not whites) between gonadal androgens and AUC for glucose, insulin, and C-peptide. Race-by-covariate interaction models reinforced the simple correlation finding. Cholesterol level was also correlated to all androgens in blacks, but not in whites. We also found that whites had higher serum triglycerides and greater AUC glucose than blacks. CONCLUSIONS: We conclude that there is a significant direct correlation between gonadal androgens and stimulated glucose, insulin, and C-peptide in blacks but not in whites. Thus, the previously reported direct correlation between gonadal hyperandrogenism and hyperinsulinemia may be a race-dependent phenomenon, hitherto an unreported observation. The implications of these findings are discussed. PMID- 10480522 TI - ACE inhibitors improve endothelial function in type 1 diabetic patients with normal arterial pressure and microalbuminuria. AB - OBJECTIVE: The purpose of this study was to test whether a short-course treatment with ACE inhibitors may restore endothelium-dependent and/or -independent vasodilation in the femoral artery of microalbuminuric patients with type 1 diabetes and normal arterial pressure. RESEARCH DESIGN AND METHODS: We studied nine normotensive microalbuminuric type 1 diabetic patients and two groups of control subjects matched for femoral artery diameter to type 1 diabetic patients after placebo (control group A, n = 17) and ACE inhibitor (control group B, n = 18) treatment, respectively. The patients were enrolled in a double-blind cross over study with a 1-week trial of either placebo, captopril (25 mg t.i.d.), or enalapril (10 mg/day) in randomized order to ascertain whether short-term ACE inhibition obtained with (captopril) or without (enalapril) a sulfhydryl donor molecule ameliorates vessel wall function. Endothelium-mediated flow-dependent vasodilation and endothelium-independent vasodilation were evaluated in the right common femoral artery by echo Doppler. RESULTS: Both captopril and enalapril normalized (control group B 22.9+/-3.2% per 8 min) endothelium-dependent response (19.6+/-7.5 and 18.0+/-5.3 vs. -10.4+/-4.1% per 8 min, P < 0.01, for both captopril and enalapril versus placebo, respectively) in the type 1 diabetic patients. Captopril (28.4+/-3.5 vs. 17.1+/-3.5% per 5 min during placebo, P < 0.05) but not enalapril (20.1+/-3.0 vs. 31.7+/-2.8% per 5 min, P < 0.05 for enalapril versus control group B, and NS for captopril vs. control group B) ameliorated endothelium-independent vasodilation in type 1 diabetic patients. CONCLUSIONS: ACE inhibition improves endothelium-dependent vasodilation in the femoral artery of normotensive microalbuminuric type 1 diabetic patients. Captopril also ameliorates endothelium-independent vasodilation, possibly through its sulfhydryl donor properties. These results may be of pathophysiological relevance to prevent cardiovascular complications in these patients. PMID- 10480523 TI - Serum levels of advanced glycation end products are increased in patients with type 2 diabetes and coronary heart disease. AB - OBJECTIVE: To investigate whether serum levels of advanced glycation end products (AGEs) and the glycoxidation product Nepsilon-(carboxymethyl)lysine (CML) are increased in patients with type 2 diabetes compared with nondiabetic control subjects and whether levels of AGEs and/or CML differ in patients with type 2 diabetes with or without coronary heart disease (CHD). RESEARCH DESIGN AND METHODS: Serum levels of AGEs and CML were measured with an immunoassay in 32 men and 21 women aged 59.3+/-6.2 years (means +/- SD) with type 2 diabetes for 7.3 + 3.1 years and in 17 men and 17 women aged 56.2+/-4.2 years without diabetes. Of the patients with diabetes, 18 had CHD. RESULTS: The serum levels of AGEs and CML were significantly increased in patients with type 2 diabetes compared with nondiabetic control subjects (median [5th-95th percentile]: AGEs 7.4 [4.4-10.9] vs. 4.2 [1.6-6.4] U/ml, P < 0.0001; CML 15.6 [5.6-29.9] vs. 8.6 [4.4-25.9] U/ml, P < 0.0001). The median level of AGEs but not CML was significantly increased in patients with type 2 diabetes and CHD compared with patients without CHD (8.1 [6.4-10.9] vs. 7.1 [3.5-9.8] U/ml, P = 0.03). There were significant positive correlations between serum levels of AGEs and CML in both patients and control subjects. CONCLUSIONS: Levels of AGEs and CML were significantly increased in patients with type 2 diabetes compared with nondiabetic control subjects, and levels of AGEs but not CML were significantly higher in patients with type 2 diabetes and CHD than in patients without diabetes. These results may indicate a role for non-CML AGEs in the development of macrovascular disease in patients with type 2 diabetes. PMID- 10480524 TI - Primary nociceptive afferents mediate the blood flow dysfunction in non-glabrous (hairy) skin of type 2 diabetes: a new model for the pathogenesis of microvascular dysfunction. AB - OBJECTIVE: To test the independent contributions of vascular endothelium, sympathetic activation and inhibition, vessel distensibility, and nociceptor mediated vasodilation in both glabrous and hairy skin circulations. RESEARCH DESIGN AND METHODS: We measured blood flow using laser Doppler techniques in 10 people with type 2 diabetes and 10 age- and BMI-matched healthy control subjects at the pulp of the index finger (glabrous skin) and the dorsum of the hand (hairy skin). A 5-min ischemic block of the arm was used to test vascular endothelium. Warming of the probe site to 45 degrees C tested neurogenic vasodilation in hairy skin only. Vessel distensibility was tested by gravitational pressure. RESULTS: Basal blood flow and reactive hyperemia did not differ between groups at either skin site. The vasodilative response to local warming (P < 0.01) and limb lowering (P < 0.05) were significantly different between groups in hairy skin but not in glabrous skin in the absence of objective measured neuropathy. Nociceptor mediated flow correlated significantly with the warm thermal threshold (r = 0.50, P < 0.05). Endothelial-mediated blood flow correlated with systolic blood pressure (r = -0.76, P < 0.01), LDL cholesterol (r = -0.62, P < 0.001), C-peptide (r = 0.65, P < 0.05), and triglycerides (r = 0.47, P < 0.05). CONCLUSIONS: These data suggest that neurogenic nociceptor-mediated vasodilation is impaired in subjects with type 2 diabetes when endothelial and sympathetic function are relatively intact. Heat-induced vasodilation may be a specific test of small heat sensitive C-fiber peripheral neurons and may be an integral part of the metabolic syndrome. PMID- 10480525 TI - Urinary excretion of albumin in adolescents with type 1 diabetes: persistent versus intermittent microalbuminuria and relationship to duration of diabetes, sex, and metabolic control. AB - OBJECTIVE: Urinary excretion of albumin is a marker for incipient diabetic nephropathy in adults. The intra-individual variability, as well as the relationship to duration of diabetes, onset of the disease, and long-term metabolic control, have not been evaluated in a large sample of pediatric patients. RESEARCH DESIGN AND METHODS: A total of 5,722 nocturnal urinary albumin excretion rates were determined in 447 children, adolescents, and young adults with type 1 diabetes, comprising 1,821 years of observation. Excretion rates were related to duration of diabetes, age at onset of diabetes, sex, blood pressure, and metabolic control. RESULTS: Based on repeated measurements in individual patients, the positive predictive value of one sample was 76%, the negative 99.5%. After a duration of diabetes of 11 years, 5% of patients displayed persistent microalbuminuria (10% after 13 years). The duration of diabetes until persistent microalbuminuria was identical for patients with prepubertal or pubertal onset of diabetes. In addition to duration, female sex (P < 0.03) and insufficient long-term metabolic control (P < 0.03) contributed significantly and independently to urinary albumin excretion. CONCLUSIONS: Determination of urinary albumin excretion rate is useful in pediatric patients. Female subjects with a long duration of diabetes and insufficient metabolic control are especially at risk for microalbuminuria. Even if persistent microalbuminuria usually becomes evident in patients aged > 11 years, the prepubertal duration of diabetes contributes equally to this risk. Good metabolic control therefore should be aspired to from the onset of diabetes. PMID- 10480526 TI - Diabetes in pregnancy and cesarean delivery. AB - OBJECTIVE: To evaluate the effect of diabetes during pregnancy on cesarean delivery and to determine whether the association between diabetes during pregnancy and cesarean delivery is mediated by birth weight. RESEARCH DESIGN AND METHODS: South Carolina 1993 birth certificates were matched through a unique identifier with infant and maternal hospital discharge records for the same year, yielding a total study population of 42,071 singleton births. Adjusted odds ratios (ORs) and 95% CIs were determined for the association between diabetes in pregnancy and cesarean delivery through multiple logistic regression, controlling for maternal age, race, education, number of prenatal care visits, length of gestation, birth weight, and a number of medical indications. RESULTS: Of the study population, 0.7% were pregnancies complicated by preexisting diabetes, 2.9% were pregnancies complicated by gestational diabetes, and 23.4% were cesarean deliveries. After controlling for confounders, including birth weight, cesarean delivery was strongly associated with both preexisting diabetes (OR [95% CI] 6.20 [4.47-8.61]) and gestational diabetes (1.71 [1.41-2.07]). The estimates remained essentially unchanged without birth weight in the model, and were substantially higher in analyses restricted to deliveries without common medical indications for cesarean delivery. CONCLUSIONS: Both preexisting and gestational diabetes increase the risk for cesarean delivery, independent of the effect of birth weight. The association is markedly greater among women without other medical indications for cesarean delivery. The increased risk of cesarean delivery for women with diabetes is mediated through other factors, which may include practice patterns and physician referrals to high-risk care. PMID- 10480528 TI - The European Association for the Study of Diabetes annual meeting, 1998: type 1 diabetes. PMID- 10480527 TI - Troglitazone: antihyperglycemic activity and potential role in the treatment of type 2 diabetes. AB - Insulin resistance is a major component of type 2 diabetes; therefore, an insulin sensitizer agent like the thiazolidinedione compound troglitazone is considered a very promising drug. Troglitazone exerts an antihyperglycemic activity in a dose dependent manner between 200 and 600 mg/day in type 2 diabetic patients treated with diet alone, sulfonylureas, or insulin. Additive antihyperglycemic effect may also be obtained by combining troglitazone and metformin. The antihyperglycemic effect of troglitazone as monotherapy is rather modest (reduction of HbA1c by 0.5 1.0%), but it appears to be somewhat greater when it is combined with other antidiabetic drugs. No double-blind studies have directly compared the activity of troglitazone with that of sulfonylureas or metformin. Troglitazone has been shown to exert additional beneficial effects on serum lipid profile and arterial blood pressure. It may be considered as a valuable alternative in insulin resistant (obese and hyperinsulinemic) diabetic patients who appear to be the best responders to the drug. However, the efficacy of troglitazone is challenged by its safety profile, and the risk of hepatotoxicity still remains a major concern in clinical practice. PMID- 10480529 TI - Sauna-induced diabetic ketoacidosis. PMID- 10480530 TI - Long-term prognosis of islet cell antibody-negative ketosis-onset diabetes with subsequent non-insulin dependency. PMID- 10480531 TI - ACE inhibitor-induced cough in hypertensive type 2 diabetic patients. PMID- 10480532 TI - Anti-HCV antibodies in a population of insulin-dependent diabetic children and adolescents. PMID- 10480533 TI - Cardiovascular risk factors in African-Americans with varying degrees of glucose intolerance. PMID- 10480534 TI - Pancreatic amyloid proteins and their relation to clinical diabetes, with special reference to serum insulin secretion. PMID- 10480535 TI - Leptin in children and adolescents with type 1 diabetes: a 2-year longitudinal study. PMID- 10480536 TI - Wegener's granulomatosis presenting with life-threatening pulmonary hemorrhage in a boy with type 1 diabetes. PMID- 10480537 TI - Blood volumes and pain following capillary punctures in children and adolescents with diabetes. PMID- 10480538 TI - Significance of parental history of type 2 diabetes on insulin sensitivity and glucose effectiveness in obese nondiabetic offspring of African-American patients. PMID- 10480539 TI - Serum fructosamine and obesity. PMID- 10480540 TI - Prediction of albumin excretion rate from albumin-to-creatinine ratio. PMID- 10480541 TI - Insulin lispro: the ideal pump insulin for patients with severe hypoglycemic unawareness? PMID- 10480542 TI - Screening for diabetic nephropathy: is measurement of urinary albumin-to creatinine ratio worthwhile? PMID- 10480543 TI - Infectious diarrhea. AB - Infectious diarrhea is an extremely common illness that affects millions of Americans annually. For most patients, the illness is a self-limited one. Its major risk is dehydration. However, for some patients, diarrhea can lead to severe dehydration or be associated with bacteremia and metastatic infection. Patients with these conditions require prompt treatment. A large number of organisms have been associated with diarrhea in humans, and most laboratories routinely screen for Salmonella, Shigella, and Campylobacter. Other bacteria, parasites, and viruses account for a significant percentage of diarrhea cases and frequently go undetected. This article summarizes many of these pathogens and describes the settings in which they can be acquired. Food distribution networks have made the delivery of previously rare foods to remote areas a commonplace occurrence; this has also led to new challenges in the diagnosis and prevention of food-borne illnesses. Outbreaks of diarrhea now frequently extend across many states. The identification of a rare strain of a bacterial pathogen or changes in the isolation rate of common pathogens may be early clues to the cause of such an ongoing outbreak. Most enteric pathogens cause disease by either stimulating the secretion of fluids at the level of the small bowel or by irritating and invading the colon. Organisms that cause disease by the latter mechanism have the potential to invade the blood stream and spread to other parts of the body, including the bones and the central nervous system. Several organisms have been associated with specific postinfectious syndromes that are responsible for additional morbidity and mortality. The antibiotic resistance of bacterial pathogens has been increasing, and this has a limiting effect on the empiric treatment choices available for suspected bacterial diarrhea. Careful attention to local sensitivity patterns and appropriate testing of the patient's isolate are among the important factors that lead to successful treatment decisions. PMID- 10480544 TI - Molecular analysis for pathogenicity of oral treponemes. PMID- 10480545 TI - Quantification of phagocytosis in human neutrophils by flow cytometry. AB - Phagocytosis represents a central element of the host response to microbial invasion. We describe a flow cytometric method for measuring the kinetics of phagocytosis of two bacteria by human polymorphonuclear leukocytes (PMNs). Over a 60-min period, isolated human PMNs were exposed to Staphylococcus aureus (rapidly phagocytosed) and Klebsiella pneumoniae (slowly phagocytosed). This method distinguished adherent from ingested bacteria by quenching fluorescein isothiocyanate-labeled extracellular bacteria with ethidium bromide. This further allowed the exclusion of dead, highly permeable, and subsequently bright-red fluorescent PMNs. Our experiments with two different bacteria, various PMN-to bacteria ratios (1:1, 1:10, 1:100), and different individuals proved that 1) flow cytometric analysis is accurate and useful for characterizing phagocytosis, 2) adherent bacteria can be distinguished from ingested bacteria after quenching with ethidium bromide, and that 3) phagocytosis kinetics of two bacteria with different onsets of phagocytosis can be determined by flow cytometry and the assessment of a score that quantifies phagocytosis. PMID- 10480547 TI - Detection of Mycoplasma fermentans in saliva sampled from infants, preschool and school children, adolescents and adults by a polymerase chain reaction-based assay. AB - Attempts were made to detect Mycoplasma fermentans in saliva sampled from 201 subjects (108 males and 93 females) aged from 4 months to 59 years by a polymerase chain reaction-based assay. M. fermentans was detected in saliva from 110 (54.7%) of 201 subjects, and 10 (28.6%) of 35 subjects aged from 4 months to 3 years. Of ten positive subjects, three were aged from 16 to 23 months and five were from 26 to 31 months. The incidence tended to increase with age up to the teens. The incidence was significantly greater in teenagers than in subjects aged from 7 to 12 years, but there was no significant difference in the incidence between the group of teenagers and each of the groups of subjects older than the teenagers. Thus, it was suggested that M. fermentans colonized the mouth at the age of about 16 months up to the age of 19 years. PMID- 10480546 TI - Cloning, sequencing, and functional expression in Escherichia coli of chaperonin (groESL) genes from Vibrio cholerae. AB - Using a series of oligonucleotides synthesized on the basis of conserved nucleotide motifs in heat-shock genes, the groESL heat-shock operon from a Vibrio cholerae TSI-4 strain has been cloned and sequenced, revealing that the presence of two open reading frames (ORFs) of 291 nucleotides and 1,632 nucleotides separated by 54 nucleotides. The first ORF encoded a polypeptide of 97 amino acids, GroES homologue, and the second ORF encoded a polypeptide of 544 amino acids, GroEL homologue. A comparison of the deduced amino acid sequences revealed that the primary structures of the V. cholerae GroES and GroEL proteins showed significant homology with those of the GroES and GroEL proteins of other bacteria. Complementation experiments were performed using Escherichia coli groE mutants which have the temperature-sensitive growth phenotype. The results showed that the groES and groEL from V. cholerae were expressed in E. coli, and groE mutants harboring V. cholerae groESL genes regained growth ability at high temperature. The evolutionary analysis indicates a closer relationship between V. cholerae chaperonins and those of the Haemophilus and Yersinia species. PMID- 10480548 TI - Modulation of barrier function of small intestinal epithelial cells by lamina propria fibroblasts in response to lipopolysaccharide: possible role in TNFalpha in inducing barrier dysfunction. AB - Recent evidence suggests an interaction between immune, enteric neural and fibroblasts in the regulation of intestinal function. Earlier, we have reported that lipopolysaccharide (LPS) induced cell proliferation, collagen synthesis and production of proinflammatory mediators in lamina propria fibroblasts. In this report, we investigated the change in transepithelial resistance (TER) as a marker of epithelial barrier function by lipopolysaccharide (LPS) and its modulation by human small intestinal lamina propria fibroblasts (HSILPF). Epithelial cells incubated with LPS alone did not show any change in the TER at any concentration or prolonged exposure. However, co-cultivation of epithelial cells with lamina propria fibroblasts which had been exposed to LPS resulted in a rapid decrease in TER by 2 hr. The decrease in the TER was continued till 8 hr followed by returning to the basal level by 24 hr. The supernatant of LPS-treated HSILPF was less effective in causing a fall in the TER than HSILPF itself. The fall in TER was accompanied by loosening of tight junctions as depicted by increased penetration of horse radish peroxidase (HRP) across the epithelial cells from the apical to the basal side. Increased incorporation of 3[H]thymidine (tritiated thymidine) in epithelial cells was observed at 48 hr in the presence of LPS-treated HSILPF. The decrease in TER during the early time period in epithelial cells was abrogated to 70% by incubating the LPS-treated HSILPF and the conditioned medium of LPS-treated HSILPF with anti-TNFalpha antibody, and not with antibody to other cytokines like IL1alpha, IL1beta, IL6 and IL8. Overall, these results suggest that TNFalpha produced by HSILPF in response to LPS as a soluble form cause a decrease in the TER and loosening of tight junctions, and such early changes in the epithelial barrier may contribute to local inflammation in the gut. PMID- 10480549 TI - Protective efficacy and immunogenicity of Vi-porin conjugate against Salmonella typhi. AB - A conjugate vaccine against Salmonella typhi was prepared by covalently binding capsular polysaccharide (Vi) with porin, both isolated from S. typhi. First, Vi and porins were extracted. The Vi was purified from S. typhi Ty2. The purified Vi conformed to the requirements of the World Health Organization. Porins were purified from S. typhi 0901. The Vi was bound to the porins by a heterobifunctional cross-linking reagent, N-succinimidyl-3-(2-pyridyl dithio) propionate (SPDP). After preparing the Vi-porin conjugate, its protective ability and immunogenicity were studied in mice following systemic immunization. The results showed that the conjugate is 6.5-fold more protective than Vi alone against S. typhi. The mice immunized with conjugate elicited higher anti-Vi antibody (IgG) levels (P < 0.01) than the mice immunized with Vi alone. Anti porin antibodies were also induced by the conjugate. To study the mucosal immune responses, secretory IgA (sIgA) in the intestinal fluid was measured. Conjugate immunized mice showed the induction of sIgA as compared to Vi alone. The results showed that when Vi is bound to porins, both isolated from same organism, the resultant conjugate induced both systemic and mucosal immune responses and provided better protection against S. typhi than Vi alone. PMID- 10480550 TI - Detection and nucleotide sequence analysis of human caliciviruses (HuCVs) from samples in non-bacterial gastroenteritis outbreaks in Hokkaido, Japan. AB - Samples of feces and vomit collected from patients during 13 non-bacterial gastroenteritis outbreaks which occurred in Hokkaido between 1995 and 1998 were examined by electron microscopy (EM) and reverse-transcription polymerase chain reaction (RT-PCR) for evidence of infection with human caliciviruses (HuCVs). In 6 food-borne outbreaks, oysters were the probable source of infection, while the origin of HuCVs was not found out for the other 7 outbreaks. One-hundred-eleven of 214 stool, vomit and oyster specimens examined gave positive results by RT PCR, while HuCVs were detected by EM in 36 of 121 stool specimens examined. We determined the nucleotide sequences of 470-bp or 373-bp PCR products amplified from the RNA polymerase region of the HuCV genomes with primer sets MR3/4 and Yuri22F/R, respectively. The sequences of different strains revealed great heterogenicity, with a range of 60 to 100% homology among strains. In a few cases, a mixed genotype was found in the same patient or same outbreak by means of nested PCR and cloning of PCR products into an appropriate vector. Of the 19 different strains found, 4 strains could be classified as Norwalk virus (genogroup 1) and the other 15 strains as Snow Mountain agent (genogroup 2) based on genotyping with homology analysis. Furthermore, the strains belonging to genogroup 2 could be classified into 4 subgroups with more than 93% homology in amino acids among strains in the subgroup. PMID- 10480551 TI - Molecular epidemiology of rabies in Thailand. AB - For the purpose of making clear the dynamics of rabies viruses that are prevalent among dogs in Asia, especially Thailand, nucleoprotein (N) genes of isolates derived from Thailand were partially sequenced, and a phylogenetic analysis was performed on the basis of the sequencing data. Firstly, all 27 isolates from Thailand belonged to one group that was distantly related to an isolate from China and was separated into at least six lineages. On the other hand, the isolate from Japan was related to viruses from the Arctic. Secondly, in order to analyze the diversity of the N gene more conveniently, restriction fragment length polymorphism (RFLP) analysis was performed on the N gene of 27 isolates from Thailand. The RFLP analysis could distinguish the lineages of each isolate, and the lineages of additional 34 isolates were deduced by this method. On examination of the geographical distribution of the six lineages, based on the results of phylogenetic and RFLP analyses, it was clear that infection cycles of the rabies virus in Thailand have tended to be maintained endemically. PMID- 10480552 TI - Correlation of T-cell response and lymphokine profile with RESA peptides of Plasmodium falciparum containing a universal T-cell epitope and an immunopotentiator, polytuftsin. AB - Two RESA repeat sequences, (EENVEHDA)2 and (DDEHVEEPTVA)2, were chemically linked to a universal T-cell epitope, CS.T3 and polytuftsin, and a natural immunopotentiator, was physically mixed with these conjugates. The immunogens were studied for in vitro antigen-induced T-cell proliferation, and cytokine levels were measured in the culture supernatants. The RESA peptide(s)-CS.T3 conjugate containing polytuftsin showed the highest stimulation index (SI) as compared to the RESA peptide-CS.T3 conjugates or RESA peptides alone. Spleen cells from mice primed with either RESA peptide(s)-CS.T3 conjugate or RESA peptide-CS.T3 conjugate containing polytuftsin, when pulsed in vitro with the respective RESA peptide, showed a higher proliferation index as compared to spleen cells primed and pulsed in vitro with the respective RESA peptides. This observation has an important relevance during natural reinfection for boosting the immune response. The culture supernatants from the cells primed and pulsed in vitro with RESA peptide-CS.T3 conjugate and RESA peptide-CS.T3 conjugate containing polytuftsin showed higher IL-2 and IFN-gamma levels as compared to the RESA peptides alone. Very low IL-4 levels were detected with the above formulations. The cytokine profile is suggestive of a CD4+ TH1 type of immune response, which is ideal for the killing of intracellular pathogens like the malarial parasite. PMID- 10480553 TI - Potentiation of immune response against the RESA peptides of Plasmodium falciparum by incorporating a universal T-cell epitope (CS.T3) and an immunomodulator (polytuftsin), and delivery through liposomes. AB - Synthetic peptides representing repeat sequences of ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum have shown poor immunogenicity and protection. In this study, the RESA peptides [(EENVEHDA)2 and (DDEHVEEPTVA)2] were chemically linked to a universal T-cell determinant, CS.T3, derived from the CS protein of P. falciparum. Polytuftsin (TKPR)40, a polymer of naturally occurring immunomodulator "tuftsin," was physically mixed with these conjugates. These preparations in alum and liposomes were immunized in four inbred strains of mice with different genetic backgrounds to study the humoral response. In the case of liposome-entrapped preparations, a 10 microg dose of antigen showed the optimum antibody response. Mice immunized with liposome containing RESA peptide(s)-CS.T3 conjugate along with polytuftsin showed the highest antibody levels in all the strains, whereas the RESA peptide(s) alone, adsorbed on alum or entrapped in liposomes, showed either poor or moderate antibody levels. The antibodies raised against liposome-entrapped preparations in both high-responder strain (SJL/J H-2s) and low-responder strain (FVB/J H-2q) showed 2 4-fold lower Kd values as compared to the alum adsorbed preparations, suggestive of high affinity antibodies. All the antigen preparations predominantly induced IgG2a and IgG2b isotype response, suggesting that the T-helper response involved is of the CD4 Thl type. The in vitro merozoite reinvasion inhibition assay showed 50-92% inhibition with sera raised against different antigen formulations. The highest percentage inhibition was observed with the RESA peptide-CS.T3 conjugate containing polytuftsin in liposomes. Thus, the incorporation of peptide antigens inside liposomes not only reduced the antigen dose by 5-fold but also elicited a high titre with high affinity antibodies and the inhibition of merozoites to RBC in vitro. Therefore, we conclude that the incorporation of these synthetic constructs in liposomes could be a useful strategy for the development of a subunit immunogen against malaria. PMID- 10480555 TI - mRNA expression of complement components and regulators in rat arterial smooth muscle cells. AB - The presence of C5b-9 complexes, some complement regulators, and abundant cytokines in atherosclerotic lesions has been reported. However, it is unclear whether these complement-associated proteins are produced by vascular smooth muscle cells (SMCs) and how they are influenced by the cytokines. In the present study, we demonstrated, by the reverse transcription-polymerase chain reaction method, the mRNA expression of complement components (C3, C4, and C5) and membrane regulators (decay-accelerating factor, membrane cofactor protein, Crry, and CD59) in cultured SMCs derived from the rat carotid artery. The expression of C9 mRNA was also induced upon stimulation by interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and/or lipopolysaccharide (LPS). Northern blot analysis showed that the mRNA expression of C3, C4, DAF and Crry was up regulated, but that of CD59 was down-regulated by IFN-gamma, TNF-alpha and/or LPS alone or by synergy. The increase of C3 mRNA by TNF-alpha or LPS and that of C4 mRNA by IFN-gamma was induced in a dose-dependent manner. The results indicate that the arterial SMCs of rat have the ability to produce complement components and regulators, which is affected by cytokines and/or LPS. Since atherosclerosis is characterized by the intimal proliferation of SMCs, the complement system including its regulators may be involved in the pathogenesis of the disease. PMID- 10480554 TI - A single cell analysis of TCR AV24AJ18+ DN T cells. AB - The T-cell receptor (TCR) BV gene of human TCR AV24+ double-negative (DN) T cells, a novel subset of natural killer (NK) T cells, was investigated by single cell sorting and single-cell polymerase chain reaction (PCR) methods. Seven of eleven TCR AV24+ DN T-cell clones utilized TCR BV8, three BV9, and one BV6. Six of seven TCR AV24/BV8+ DN T-cell clones had identical TCR beta and alpha chains, indicating that they were the same clone. All three TCR AV24/BV9+ DN T-cell clones also demonstrated the same amino acids in the CDR3 region. These findings strongly suggest that the usage of TCR beta and alpha chains on TCR AV24+ DN T cells is extremely restricted, supporting the notion that these cells recognize highly limited T-cell epitopes on antigens. All TCR AV24+ clones expressed the NKR-P1A mRNA, and so were true NK T cells. IL-2 and IL-4 mRNAs were detected in all clones, suggesting that the majority of these cells were Th0-type T cells. Six clones overexpressed Fas-ligand (Fas-L) mRNA and Fas antigen was detected on all clones at the mRNA level. In conclusion, TCR AV24+ DN T cells might recognize restricted T-cell epitopes on antigens and function as Th0-type T cells, inducer cells to Th1- or Th2-type T cells (regulatory T cells), and as Fas-L-positive cytolytic T cells. PMID- 10480557 TI - Isolation of feline parvovirus from peripheral blood mononuclear cells of cats in northern Vietnam. AB - Feline parvovirus (FPV) was isolated rather frequently from the peripheral blood mononuclear cells (PBMCs) of cats in northern Vietnam by coculturing with MYA-1 cells (an interleukin-2-dependent feline T lymphoblastoid cell line) or Crandell feline kidney (CRFK) cells (a feline renal cell line). Efficiency of virus isolation was higher in MYA-1 cells than in CRFK cells. Interestingly, among the 17 cats from which FPV was isolated, 9 cats were positive for virus neutralizing (VN) antibody against FPV, indicating that FPV infected PBMCs and was not eliminated from PBMCs even in the presence of VN antibodies in the cats. PMID- 10480556 TI - The majority of lymphocytes in the bone marrow, thymus and extrathymic T cells in the liver are generated in situ from their own preexisting precursors. AB - Parabiotic pairs of B6.Ly5.1 and B6.Ly5.2 mice were used to investigate how lymphocytes in various organs and various lymphocyte subsets mixed with partner cells. The origin of partner cells was determined by using anti-Ly5.1 mAb in conjunction with immunofluorescence tests. Parabiosis was also produced after the irradiation of B6.Ly5.2 mice at various doses to prepare an immunosuppressive partner. Irrespective of irradiation, lymphocytes and other hematopoietic cells in the bone marrow and lymphocytes in the thymus showed a low mixture of partner cells in comparison with those of all other organs tested. On the other hand, lymphocytes in the blood, spleen, and lymph nodes became a half-and-half mixture of their own cells and partner cells by 14 days after parabiosis. Among lymphocyte subsets, intermediate CD3 cells (i.e., CD3int cells) and NKT cells (i.e., NK1.1+ subset of CD3int cells) in the liver also showed a low mixture of partner cells. The present results raise the possibility that lymphocytes in the bone marrow and thymus, and extrathymic T cells in the liver might be in situ generated from their own preexisting precursor cells. Another observation was that, after irradiation, partner cells showed accelerated mixture even if they showed a low mixture under non-irradiated conditions. However, only lymphocyte subsets with the same phenotype as those of preexisting cells entered the corresponding sites. PMID- 10480558 TI - Microsurgical reconstruction of the through-and-through defect in head and neck cancer: is it worth it? AB - As health resources diminish, there are compelling reasons to utilize health dollars in a fiscally responsible manner. The reconstruction of complex oromandibular defects involving mucosa, bone, and skin coverage poses one of the greatest challenges in microsurgery of the head and neck. The cancer patient who requires a through-and-through resection and microsurgical reconstruction usually has a poor prognosis. In this study, the authors examine whether this type of surgery is worthwhile in terms of cost, functional outcome, and patient satisfaction. Of 16 cases of through-and-through oromandibular reconstruction performed, the survival outcome of ten (n = 10) advanced cases requiring immediate oromandibular reconstruction (7 radial forearm flaps; 3 scapular flaps) is presented. Six cases were considered cured and required delayed reconstruction. Seven of the 10 patients died within 39 months postoperatively, while three survived up to 68 months postoperatively. The combined experience of these ten patients was examined using the Kaplan-Meier (product-limit) estimator of the survival curve. Results show that of the seven patients who died of disease, five did so within the first postoperative year. More important, among those five patients who survived for more than one postoperative year, three were still alive up to 68 months, representing a combined total of over 15 postoperative years. The probability of long-term survival is good in through-and through oromandibular cancer patients who can survive to 1 year postoperatively, and it is proposed that microsurgical reconstruction, albeit costly, remains a worthwhile procedure, PMID- 10480559 TI - Combined pedicle and free-tissue transfer to improve functional restoration of the foot: the "backpack principle". AB - Reconstruction of weight-bearing areas of the foot remains a challenging problem for the reconstructive surgeon. The principle described involves the combination of an innervated regional pedicle flap providing sensate, durable coverage of the weight-bearing heel, and a microvascular fascia flap for thin, pliable coverage of a large surface defect, based on the same vascular bundle. This provides excellent long-term stability and minimizes the disruption of blood supply to the foot. The result after 3 years demonstrates that this concept may represent an interesting solution for these particular problems. PMID- 10480560 TI - Superficial ulnar artery: embryology, case report, and clinical significance in reconstructive microsurgery. AB - In this paper the authors describe and illustrate a rare anatomic variation of the ulnar artery, a superficial ulnar artery. The possible embryologic events responsible for this variation are discussed. In addition, they describe the surgical considerations that should be kept in mind when this abnormality is encountered in the clinical setting of free-tissue transfer and vascularized nerve grafting. PMID- 10480561 TI - Postoperative irradiation: are there long-term effects on nerve regeneration? AB - This study investigates the histomorphologic and functional effects of external beam irradiation on peripheral nerve regeneration in the rat model. All animals received a 1.5-cm interposition isogeneic nerve graft to the right posterior tibial nerve. Group 1 animals served as controls and Groups 2 and 3 received fractionated postoperative irradiation doses of 66 and 106 Gy, respectively. All animals were evaluated for functional recovery with monthly walking-track analysis. At the end of 8 months, the animals were sacrificed and segments of the grafted and distal nerve were harvested for histomorphologic analysis. A statistically significantly fewer number of axons were found in the distal segment of the irradiated specimens, compared to controls. There was no significant difference in the nerve-fiber density of the grafted or distal segments, compared to controls. Functional evaluation by walking-track analysis showed no difference between the irradiated groups and controls over the length of the study. These results suggest that acute nerve grafting of nerve defects in the face of planned postoperative irradiation is safe. PMID- 10480562 TI - Using a direct current electrical field to promote spinal-cord regeneration. AB - The authors used a direct current electrical field to promote spinal-cord regeneration in a canine model. Thirty-two dogs were randomly divided into four groups. Complete spinal-cord injury was induced, and electrical stimulators were then placed in the animals. Group 1 served as controls; Groups 2 to 4 were experimental groups, with varying stimulator voltages: 0V in Group 1, 12V in Groups 2 and 4, and 6V in Group 3, with the stimulator implanted 6 hr after spinal-cord injury in Group 4. Functional, electrophysiologic and morphometric assessments were carried out 1 to 3 months postoperatively. Results showed that spinal-cord function, cortical somatosensory evoked potentials, number of neurons, sectional area of neurons, and Nissl body density in the experimental groups were much better than those in the control group. In addition, all the indices in Group 2 were better than those in Groups 3 and 4. This indicated that direct current electrical stimulation could effectively promote spinal-cord regeneration and functional recovery in this model. The 12V voltage was safe for the animals. The stimulator was not rejected by the host for a relatively long period of time. PMID- 10480563 TI - Quadriceps femoris muscle flap: largest muscle flap model in the rat. AB - The purpose of this paper is to present a new muscle flap in the rat: the quadriceps femoris muscle flap based on a pedicle consisting of the femoral vessels. In order to establish the anatomic details of this model, seven rats were explored bilaterally, and the regional anatomy of the thigh was examined. The technical aspect of the model was established by the unilateral harvesting of 18 quadriceps femoris muscles. Findings were that this muscle is primarily supplied by a muscular branch originating at the superficial circumflex iliac artery. The average muscle weight was 6 g and the average pedicle length with femoral vessels was 6 mm. Eight of the harvested flaps were transplanted to the contralateral thigh, and the pedicle was anastomosed to the femoral vessels. The other ten flaps were resutured back to their beds. At 72 hr postoperatively, all flaps were viable with the exception of one of the transplanted flaps which was found to be necrotic. The quadriceps femoris muscle flap is technically both a reliable and simple model. With an average weight of 6 g, this flap is by far the largest described in the rat, and offers a convenient model for testing flap related techniques and outcomes. PMID- 10480564 TI - Vascular effects of epinephrine, lisinopril, and chlorpromazine in diabetic and non-diabetic rats. AB - In this study, the vascular responses of diabetic rat femoral arteries to epinephrine were investigated. The effects of lisinopril (ACE inhibitor) on vascular epinephrine sensitivity were also tested in a different group. This study was carried out in sodium pentobarbital-anesthetized rats 8 weeks after induction of diabetes with streptozotocin. After extensive dissection of the femoral arteries with adventitial stripping, epinephrine and chlorpromazine were applied to the vascular wall, and their vascular effects were compared in streptozotocin-diabetic (STZ-D), lisinopril-administered streptozotocin-diabetic (LASTZ-D), lisinopril-administered nondiabetic (LAND), and non-diabetic (ND) groups. Vasoconstriction was induced by epinephrine in all groups in a dose response fashion. There were statistically significant differences in maximum percent constriction between STZ-D and LASTZ-D groups. There was also a significant increase in sensitivity to epinephrine in the STZ-D group. The vasoconstriction induced by epinephrine was relieved by chlorpromazine in all groups. Results suggest that there are important functional abnormalities in the responses of vessels to epinephrine in diabetics, and that the attenuation of vasoconstriction by ACE inhibitors may have beneficial effects in microsurgical procedures performed on diabetic patients. Topically-applied chlorpromazine appears to be effective in relieving vasospasm due to epinephrine, and may be a useful tool to resolve perioperative vascular spasm in microsurgical procedures for diabetic and non-diabetic patients. PMID- 10480565 TI - Choline acetyltransferase activity in collateral sprouting of peripheral nerve after surgical intervention: experimental study in rats. AB - The purpose of this study was to establish an assay of choline acetyltransferase (ChAT) activity to investigate the regeneration of injured peripheral nerve, repaired by end-to-end or end-to-side neurorrhaphy. Murine sciatic and peroneal nerves were exposed, and the peroneal nerve was transected at a site 5 mm from its ramification. For end-to-side neurorrhaphy, an epineurotomy producing a 5-x5 mm window was carried out on the tibial nerve, just above the level of gastrocnemius muscle ramification. The peroneal nerve stump was then sutured end to-side to the tibial nerve window. For end-to-end neurorrhaphy, the peroneal stump was directly sutured end-to-end. ChAT activity was measured at a site distal to the peroneal stump at 1 to 3 months postoperatively, and the results were compared among four groups: 1) end-to-end neurorrhaphy group; 2) end-to-side neurorrhaphy group; 3) unrepaired group; and 4) positive controls. ChAT activity in the end-to-side neurorrhaphy yielded approximately two-thirds the value of the end-to-end neurorrhaphy, and more than half the value of positive controls at 3 months postoperatively. Histologic sections of the end-to-side and end-to-end sutured peroneal nerve demonstrated large numbers of myelinated axons and Schwann cells at the third postoperative month. All the results demonstrated that end-to side neurorrhaphy is comparable to well-performed end-to-end neurorrhaphy, thus providing another option for surgical treatment of avulsion nerve injury and massive nerve defect. PMID- 10480566 TI - End-to-side neurorrhaphy: evaluation of axonal response and upregulation of IGF-I and IGF-II in a non-injury model. AB - This research group has introduced a model of end-to-side neurorrhaphy, in which reinnervation occurs without frank damage to donor axons. The current study used in situ hybridization to test the hypothesis that insulin-like growth factor (IGF I and IGF-II) mRNA levels increase at the coaptation site and grafted nerve following end-to-side repair, and that this increase is associated with axonal sprouting and growth. One week after end-to-side coaptation, IGF-I mRNA was localized predominantly on the epineurial side of the graft perineurium, while IGF-II was seen mainly on the endoneurial side. IGF-I hybridization was greatest at this time and declined by 2 weeks post-procedure. No changes in IGF mRNA levels occurred in the distal donor nerve. The increase in IGF-I mRNA at 1 week preceded the appearance of myelinated axons. The presence of myelinated axons within the graft 2 weeks after end-to-side coaptation was associated with a decline in IGF-I mRNA. These data are the first to demonstrate increased IGF mRNA levels associated with axonal sprouting and growth following end-to-side neurorrhaphy. Moreover, the findings support those of earlier studies by others implicating IGFs in axonal regeneration. The increase in IGF mRNA during sprouting and axonal growth into an end-to-side coaptation indicates that the local therapeutic augmentation of endogenous IGF levels at the coaptation site may enhance axonal sprouting from a minimally injured donor nerve, and thereby increase the number of axons that reinnervate the graft. PMID- 10480567 TI - High-mannose type oligosaccharide-dependent apoptosis in U937 cells induced by pradimicin, a mannose-binding antibiotic. AB - Cell surface oligosaccharides play a role in a variety of biological events such as cell adhesion and signal transduction. We have shown that BMY-28864, a semi synthetic analog of pradimicin, induced apoptosis of U937 cells which had been incubated with 1-deoxymannojirimycin, an inhibitor of mannosidase I. BMY-28864 was not cytotoxic to the cells which had been cultivated with other glycosidase inhibitors such as castanospermine and swainsonine. We thus propose that BMY 28864 induces apoptosis by acting on a specific mannose-rich oligosaccharide, presumably (Man)9(GlcNAc)2+. PMID- 10480568 TI - Involvement of Ca2+ ion and reactive oxygen species as a mediator in pradimicin induced apoptosis. AB - Pradimicin (PRM) induces apoptosis in mammalian cells which had been incubated with 1-deoxymannojirimycin (DMJ). Flow cytometric analysis revealed that PRM preferentially induced apoptosis to the cells of the G1 phase. Two possible mediators in this apoptotic cascade were identified. Exposure of DMJ-treated cells to PRM resulted in a rapid (approximately 5 seconds) and slow (approximately 30 minutes) elevation of the intracellular calcium level. Reactive oxygen species (ROS) were proved to be involved in this system by the fact that the apoptosis was completely inhibited by treating the cells with a ROS scavenger, N-acetylcysteine in prior to the PRM stimulation. PMID- 10480569 TI - Topostatin, a novel inhibitor of topoisomerases I and II produced by Thermomonospora alba strain No. 1520. III. Inhibitory properties. AB - A novel inhibitor of topoisomerases designated as topostatin was isolated from the culture filtrate of Thermomonospora alba strain No. 1520. The inhibitory activity of topostatin was shown to be pH- and temperature-dependent with a maximum around at pH 6 and 28 degrees C. The stability of topostatin decreased with decreasing pH and rising temperature. Topostatin inhibited topoisomerases I and II in a competitive manner with respect to DNA. The inhibitor also inhibited some restriction endonucleases such as Sca I, Hind III and Pst I, but not Alu I, Bam HI, Eco RI, RNase A, DNase I, DNase II and DNA ligase. Topostatin did not induce the nuclear accumulation of p53 protein by DNA damage in the normal human cells. PMID- 10480570 TI - Sanglifehrins A, B, C and D, novel cyclophilin-binding compounds isolated from Streptomyces sp. A92-308110. I. Taxonomy, fermentation, isolation and biological activity. AB - A novel class of macrolides for which the name sanglifehrins is proposed, has been discovered from actinomycete strains based on their high affinity binding for cyclophilin A (CypA), an immunophilin originally identified as a cytosolic protein binding cyclosporin A (CsA). The sanglifehrins were produced by Streptomyces sp. A92-308110. They were isolated and purified by extraction and several chromatographic, activity-guided steps. Sanglifehrins A and B exhibit a 10 to approximately 20 fold higher affinity for CypA than CsA, whereas the affinity of sanglifehrins C and D for CypA is comparable to that of CsA. Sanglifehrins exhibit a lower immunosuppressive activity than CsA when tested in the mixed lymphocyte reaction. Their in vitro activity indicates that they belong to a novel class of immunosuppressants. PMID- 10480571 TI - Sanglifehrins A, B, C and D, novel cyclophilin-binding compounds isolated from Streptomyces sp. A92-308110. II. Structure elucidation, stereochemistry and physico-chemical properties. AB - A novel class of macrolides, the sanglifehrins, was discovered by screening of actinomycete strains with a cyclophilin-binding assay. The chemical structures and absolute stereochemistries of the sanglifehrins A, B, C and D were determined unambiguously by NMR-techniques and by X-ray crystallography of the complex with cyclophilin A. Sanglifehrin A consists of a 22-membered macrocycle containing a tripeptide subunit and features in position 23 a chain of nine carbon atoms bearing a spirocyclic substituent. Sanglifehrins A and B are genuine metabolites whereas sanglifehrins C and D are artefacts. PMID- 10480572 TI - UK-2A, B, C and D, novel antifungal antibiotics from Streptomyces sp. 517-02. IV. Comparative studies of UK-2A with antimycin A3 on cytotoxic activity and reactive oxygen species generation in LLC-PK1 cells. AB - UK-2A, a novel antifungal antibiotic, is a structural relative of antimycin A3 (AA) and its mode of action is similar to that of AA which inhibits mitochondrial electron transport at complex III. In spite of their structural resemblance, AA had strong cytotoxicity while UK-2A had little cytotoxicity against LLC-PK1 cells as well as other types of cultured cells. When cells were treated with UK-2A or with AA the intracellular ATP content decreased significantly within 5 minutes in glucose-free medium to almost the same extent in both cases. Moreover, under the same conditions, UK-2A killed cells at a similar rate to AA. This suggested that UK-2A entered into the cells and, like AA, inhibited mitochondrial electron transport. On the other hand, AA stimulated reactive oxygen species (ROS) production within 5 minutes even at a low concentration of 1 microM whereas UK-2A did not show such an effect. The difference in the ROS-producing abilities of UK 2A and AA may account for the different cytotoxic effects of the two compounds. PMID- 10480573 TI - Effect of thiazinotrienomycin B, an ansamycin antibiotic, on the function of epidermal growth factor receptor in human stomach tumor cells. AB - Thiazinotrienomycin B (TT-B), an ansamycin isolated from fermentation broths of Streptomyces sp. MJ672-m3, inhibited the growth in vitro of human stomach tumor SC-6 cells over 10 times more strongly than the growth of other human tumor cells, such as HeLa (cervix), T24 (bladder) and LX-1 (lung). The extent of growth inhibition by TT-B of SC-6, but not of LX-1 nor T24, was lowered in a competitive manner by raising serum concentrations in the culture medium. TT-B inhibited the cell cycle progression of SC-6 at an early stage of the progression from G0/G1 to S. The inhibition was again competitive with serum concentrations in the culture medium. No direct inhibition of DNA synthesis was observed at the concentration range which caused the cell cycle arrest. TT-B and anti-epidermal growth factor receptor (anti-EGFR) were antagonistic to each other in inhibiting the cell cycle progression of SC-6 from G0/G1 to S, suggesting that the two compounds share the same target, EGFR. The kinase activity of EGFR was little inhibited by TT-B in a cell-free system. PMID- 10480574 TI - AS-924, a novel bifunctional prodrug of ceftizoxime. AB - To improve the oral absorption of ceftizoxime (CZX), 7beta-[(Z)-2-(2-aminothiazol 4-yl)-2-methoxyiminoacetamido]- 3-cephem-4- carboxylic acid, we synthesized and evaluated a novel series of bifunctional prodrugs, in which L-alanine was introduced into the aminothiazole-oxime moiety at the C-7 position of the various lipophilic esters of CZX. Among these prodrugs, pivaloyloxymethyl 7beta-[(Z)-2-(2 (S)-alanylaminothiazol-4-yl)-2-methoxyiminoa cetamido]-3-cephem-4-carboxylate hydrochloride (ceftizoxime alapivoxil, AS-924) was well absorbed after oral administration in experimental animals and showed potent therapeutic effects in mice infected with gram-positive and gram-negative bacteria. PMID- 10480575 TI - New members of the macrosphelides from Microsphaeropsis sp. FO-5050 IV. PMID- 10480576 TI - Novel antibiotics pyrisulfoxin A and B produced by Streptomyces californicus. PMID- 10480578 TI - The specialty of allergy/immunology is not overrepresented in the physician workforce. PMID- 10480577 TI - A novel screen for the detection of chitin acting antifungal compounds. PMID- 10480579 TI - Environmental exposure unit: a system to test anti-allergic treatment. AB - LEARNING OBJECTIVES: Reading this article will enable readers to recognize the Environmental Exposure Unit (EEU), its historic development and its current role as a system to test anti-allergic treatment; to recognize clinical relevance of this test system and its relationship with other pollen challenge methods of evaluation of anti-allergic medication; and, to recognize variables associated with standard clinical studies of anti-allergic medication. Readers will review four studies of antihistamines tested in the Environmental Exposure Unit, three studies on nasal corticosteroids, one on topical eye drops and one on immunotherapy conducted in the EEU. DATA SOURCES: The EEU has been in operation since 1985 preceded by a prototype challenge system to assess respiratory effects of urea formaldehyde foam insulation. A number of studies on the onset of action and efficacy of different antihistamines and nasal corticosteroids as well as other treatments have been completed producing accurate and consistent results influenced to some extent by study designs. STUDY SELECTION: Studies of commonly used antihistamines and nasal corticosteroids are discussed in detail and represent several of the studies undertaken to date in the EEU. RESULTS: Controlled ragweed pollen exposure using the EEU has shown that some antihistamines demonstrate an onset of action within 30 minutes while others have taken up to 3 hours to produce significant effect. Nasal corticosteroids evidenced the onset of clinical improvement at 5 to 6 hours with significance over placebo between 6 and 12 hours depending on dose. CONCLUSION: The EEU is an effective pollen delivery system that accurately and consistently determines the onset of action and efficacy of anti-allergic treatment in large groups of subjects. It eliminates variables associated with various other methods of evaluation of these medications but does not supplant the need for such evaluations. PMID- 10480580 TI - Chronic idiopathic(?) urticaria: a satisfactory outcome. PMID- 10480581 TI - The future physician workforce: an allergy-immunology specialist perspective. PMID- 10480582 TI - Comparison of ipratropium bromide 0.03% with beclomethasone dipropionate in the treatment of perennial rhinitis in children. AB - OBJECTIVE: To compare the safety and efficacy of ipratropium bromide 0.03% (IB) with beclomethasone dipropionate 0.042% (BDP) in the treatment of perennial rhinitis in children. METHODS: Thirty-three children with nonallergic perennial rhinitis (NAPR) and 113 with allergic perennial rhinitis (APR) were randomly assigned to either IB or BDP for 6 months in a single-blind, multicenter protocol in which the physician was blinded to treatment. At each visit, patients and physicians rated symptom control of rhinorrhea, nasal congestion, and sneezing. Patients also completed quality of life questionnaires at baseline and after 6 months of therapy. RESULTS: Both treatments showed a significant improvement in control of rhinorrhea, congestion, and sneezing compared with baseline over the 6 months of treatment (P < .05). Only for the control of sneezing was BDP consistently better than IB (P < .05). Among the patients given IB, 61% to 73% assessed the control of rhinorrhea as good or excellent on different study visit days, 43% to 60% similarly rated the control of nasal congestion, and 39% to 43% the control of sneezing. The results for BDP were 68% to 78% for the control of rhinorrhea, 55% to 72% for the control of nasal congestion, and 54% to 68% for the control of sneezing. Quality of life assessment documented that both drugs significantly reduced interference with daily activities and disturbance of mood due to rhinorrhea compared with baseline (P < .05). Both treatments were well tolerated with IB causing less nasal bleeding and irritation than BDP. CONCLUSIONS: Ipratropium bromide was safe and effective in controlling rhinorrhea and diminishing the interference by rhinorrhea in school attendance, concentration on school work, and sleep. Ipratropium bromide was as effective as BDP in the control of rhinorrhea and showed a relatively good effect on congestion. Patient and physician assessment favored BDP in the control of sneezing. PMID- 10480583 TI - Prevalence and burden of illness for asthma and related symptoms among kindergartners in Chicago public schools. AB - BACKGROUND: Asthma mortality rates in poor communities of Chicago are among the highest in the country. Possible explanations include increased asthma prevalence, increased severity, and suboptimal health care. OBJECTIVE: To estimate the prevalence of asthma and asthma-related symptoms among inner-city kindergarten children, and to characterize their burden of illness, asthma related health care access, and pharmacologic treatment. METHODS: Cross-sectional survey of parents of kindergartners was conducted in 11 randomly selected Chicago elementary schools. A self-administered 16-item questionnaire was given to parents of kindergartners. Parents who reported doctor-diagnosed asthma or at least one of several key asthma-related symptoms were then interviewed with a supplemental questionnaire examining asthma-related health care and medication use. RESULTS: Based on data from 638 children [mean age 5.7 (SD = 0.6) years], the prevalence of diagnosed asthma was 10.8%. Sixteen percent of the respondents reported that their child had wheezed in the past year. The prevalence of asthma related symptoms unassociated with a diagnosis of asthma was 30.1%. The children with diagnosed asthma had evidence of a high burden of illness: over 40% were reported to have had sleep disturbance due to wheezing > or =1 to 2 nights/week and 86.6% reported acute care visits for respiratory symptoms in the past year. Self-reported access to medical care was high. Over 40% of the children with doctor diagnosed asthma were reported to have used a beta2-agonist in the preceding 2 weeks, and 12.2% used an inhaled anti-inflammatory. CONCLUSIONS: These data suggest that asthma prevalence in school-aged children in inner-city communities may be higher than US estimates. The burden of illness experienced by these children is substantial. Also, a large proportion of children were reported to have respiratory symptoms consistent with asthma, and no asthma diagnosis, suggesting possible undiagnosed asthma. While measures of health care access appear to indicate that the majority of children with asthma experience no identified barriers to health care, there is evidence to suggest undertreatment. PMID- 10480584 TI - Comparative study of extended release albuterol sulfate and long-acting inhaled salmeterol xinafoate in the treatment of nocturnal asthma. AB - BACKGROUND: Nocturnal worsening of asthma is a common problem in asthma and is associated with increased morbidity and mortality. Long acting beta-2 agonists are considered long-term symptom control medications, especially for nocturnal symptoms. OBJECTIVE: To compare efficacy of an extended release oral beta-2 agonist, albuterol sulfate (Volmax), to a long-acting inhaled agent, salmeterol (Serevent) in the treatment of nocturnal asthma. METHODS: This was a multicenter double-blind, double-dummy, randomized, crossover design with a 1-week baseline period and two 3-week treatment periods separated by a 7 to 9-day washout. An optional 2-week, open-label phase was conducted to evaluate patient preference. RESULTS: A total of 46 patients were included in the efficacy analysis. For the primary outcome variable of morning peak expiratory flow, there were similar and significant improvements over the 3-week treatment period for both medications compared with baseline (P < .001). Similar improvements were seen in the overnight change in PEF values (P < .001). The morning and overnight changes in FEV1 were not significantly different between treatment arms (P > .05). There were significant improvements in both treatment periods in regard to the percentage of nights without awakenings (baseline 53.6+/-5.3%), extended release albuterol 83.3+/-3.0% (P < .001), and salmeterol 88.8+/-2.4%. The percentage of patients who had no awakenings during treatment did not differ significantly for the two medications. Both treatments also resulted in a decrease in the use of rescue albuterol (extended release 2.66+/-0.35 puffs per day, salmeterol 1.85+/ 0.29) from baseline (4.57+/-0.41, P < .001). There was a significant difference between groups (P = .001). The reasons why patients preferred one medication over the other varied. CONCLUSION: Both extended release albuterol tablets and inhaled salmeterol resulted in similar bronchodilation and good control of nocturnal asthma symptoms. PMID- 10480585 TI - Eosinophils isolated by magnetic cell sorting respond poorly to lipid chemoattractants. AB - BACKGROUND: Eosinophils play an important role in allergic inflammation. In vitro methods to isolate human eosinophils for the study of chemotactic responses are essential in understanding the mechanisms involved in tissue eosinophilia. OBJECTIVE: We compared LTB4 and PAF-induced chemotactic responses of eosinophils isolated by the standard Percoll (positive selection) versus the magnetic cell separation systems (MACS) (negative selection) technique. METHODS: Discontinuous Percoll gradients were preceded by dextran and Ficoll-Paque steps, and followed by gelatin wash and red blood cell (RBC) lysis. MACS isolation included Percoll 1.090 g/mL layering and RBC lysis; incubation with CD16 antibody conjugated to magnetic beads (to bind neutrophils); and isolation of eluate from column positioned in magnet. RESULTS: Percoll-isolated eosinophils migrated to the lipid mediators, LTB4 and PAF, in a dose-responsive fashion. Although MACS isolation provided a greater number and higher purity of eosinophils, these eosinophils migrated less to LTB4 and PAF. Neither dextran sedimentation, dextran and Ficoll Paque, nor dextran Ficoll-Paque and Percoll prior to MACS isolation reversed the decreased chemotactic responses observed with MACS isolated eosinophils. Further, Percoll-isolated eosinophils further purified with CD16 MicroBeads did not respond as well to LTB4 or PAF. CONCLUSIONS: The technique used to isolate eosinophils clearly affects the chemotactic responsiveness of this cell to LTB4 and PAF. Since several in vivo studies suggest that LTB4 and PAF are eosinophil chemoattractants, Percoll isolation of these cells might be more appropriate for studies involving eosinophil chemotactic responses to these lipid mediators. PMID- 10480586 TI - Systemic reactions to allergy skin tests. AB - BACKGROUND: Skin testing is a common diagnostic tool in allergy. It is considered a safe procedure, although systemic reactions have been reported. OBJECTIVE: To identify the systemic reaction rates of allergy skin tests and to determine the clinical outcome of such reactions. METHOD: This retrospective study used a computerized database at the Mayo Clinic to identify patients who developed systemic reactions to skin tests. Altogether 497,656 skin tests were performed on 18,311 patients from January 1992 to June 1997. Skin puncture tests were performed on 16,505 patients. Skin puncture and intradermal skin tests were performed on 1,806 patients. Systemic reactions were evaluated and treated by physicians. RESULTS: There were 6 systemic reactions, an overall rate of 33 systemic reactions per 100,000 skin tests. All six patients had asthma. The systemic reaction rates for latex skin testing was 152 or 228 reactions per 100,000 latex skin tests, to penicillin and antibiotics 72 reactions per 100,000 penicillin and antibiotics skin tests, and to aeroallergens 15 or 23 reactions per 100,000 aeroallergen skin tests. The systemic reaction rate for skin puncture test was 30 reactions per 100,000 skin puncture tests, for skin puncture and intradermal skin tests, the rate was 55 reactions per 100,000 skin puncture and intradermal skin tests. All 6 patients were treated and dismissed within 1 hour after treatment. CONCLUSION: The systemic reaction rate to skin tests was very low. Systemic reactions were mild and all patients recovered fully within 1 hour. PMID- 10480587 TI - Effect of tree and grass pollens and fungal spores on spring allergic rhinitis: a comparative study. AB - BACKGROUND: Current literature indicates tree pollen is less important than grass pollen as the cause of spring allergic rhinitis. The role of fungal spores is secondary and uncertain. OBJECTIVE: To assess the relative allergenicity of tree and grass pollen and fungal spores as the cause of spring allergic rhinitis in central Indiana. METHODS: (1) Chart review of 189 Indiana natives with seasonal allergic rhinitis to ascertain clinical sensitivity to tree or grass pollens. (2) A prospective study conducted in the spring of 1997 in 51 patients, correlating patients' symptom and medication scores with daily pollen and spore counts, obtained with a Burkard spore trap. RESULTS: One hundred thirty of 189 patients (69%) reported symptoms before May with positive prick test to at least one tree pollen. One hundred thirty-four patients (71%) had symptoms in May and June with positive test to grass pollen. The ratio between the two groups is 1:1, in contrast to a ratio of 1:4 derived from the literature. The prospective study revealed a symptom score increase parallel to the rise of tree pollen counts beginning in mid-March and reaching a plateau in early May prior to the onset of grass season. Medication use continued to increase and peaked at the height of grass pollination. Spore counts increased in late May and stayed at high levels throughout June, yet symptom and medication scores steadily declined following the peak of grass pollination in early June. CONCLUSIONS: This study suggests that in central Indiana, tree pollen is more important as the cause of spring allergic rhinitis than that suggested by the literature. This study confirmed previous observations that fungal spores are substantially less allergenic than pollens. PMID- 10480588 TI - Lymphocyte membrane antigen expression and intracellular cytokine patterns in an asymptomatic patient with persistently high serum levels of IgE. AB - BACKGROUND: Patients completely asymptomatic with extremely high levels of IgE have rarely been reported. One such case, in which the immunophenotype pattern of lymphocyte subsets and their cytokine profile were investigated, is described here. OBJECTIVE: To assess whether the cytokine production was consistent with a T helper 2-type immune response, as suggested by theories regarding the functional polarization of helper and cytotoxic T cells in hyper-IgE conditions. METHODS: An asymptomatic 79-year-old man presented with persistent high levels of serum IgE and sporadic hypereosinophilia without any evidence of an underlying pathologic condition. We investigated the immunophenotype of circulating lymphocytes, the expression/release of CD30 (a member of the tumor necrosis factor receptor family preferentially associated with T helper 2-type immune responses) and the intracellular patterns of interferon-gamma (IFN-gamma), Interleukin-2 (IL-2), IL-4, IL-5, and IL-10 production by T cell subsets, as evaluated by single-cell flow-cytometric analysis. RESULTS: The majority of lymphocytes displayed the membrane immunophenotype of NK cells. Both CD4+ and CD8+ T cells were reduced and expressed the "memory" (CD4+/CD45RO+) and the "naive" (CD8+/CD45RA+) phenotypes, respectively. Among CD4+ T cells, CD30 expression was increased in the resting condition and was further inducible following stimulation with mitogenic anti-CD3. Interleukin-4, IL-2, and IL-10 production by CD4+ T cells was increased, whereas IFN-gamma was reduced as compared with normals. CONCLUSIONS: The data suggest that a polarization of CD4+ T cells towards a T helper 0/2-type cytokine pattern occurred in this patient in spite of CD4+ cell reduction and NK cell expansion. PMID- 10480589 TI - Specific IgE antibodies to aeroallergens and food among Israelis. AB - BACKGROUND: There are no data about specific IgE antibodies to food and aeroallergens in the Israeli population. OBJECTIVE: We sought to determine the relationship between selected demographic parameters and rate of sensitivity to various allergens throughout all ages. METHODS: Analysis was carried out on 776 positive specific IgE from 655 patients (347 to food, 429 to aeroallergens, and 121 to both). The studied parameters were age and gender. RESULTS: Age-related analysis showed the highest positivity to food (eggs, legumes, and milk) and the lowest to pollens during infancy. High sensitivity to house mites prevailed at all ages. Sensitivity to pollens and epidermals increased with age while sensitivity to eggs, legumes, and milk decreased. Above the age of 30 years, the rate of sensitivity to all tested aeroallergens and food was similar and close to the average value. Our data showed a significant cross-sensitivity between various foods and aeroallergens and also confirmed cross-sensitivity results reported earlier. CONCLUSIONS: (1) Age is a dominant factor affecting type I hypersensitivity. House mite is the most common allergen throughout life. (2) The age distribution and nature of sensitivity to allergens in Israel has some similarities to Spain but differ from other countries. (3) Cross-sensitivity between food and aeroallergens was found. PMID- 10480590 TI - Evaluation of IgE-sensitization to fungi in HIV-positive patients with eczematous skin reactions. AB - BACKGROUND: Human immunodeficiency virus infection is associated with declining immune function and polyclonal B-cell activation leading to elevated IgE-levels. In selected patient categories, increased total IgE may be associated with allergic diseases. Furthermore, a significant number of patients with low CD4+ cell numbers have various skin manifestations, eg, eczema and dermatophytosis. Patients with chronic fungal infections and a tendency to produce increased levels of specific IgE may become allergic and IgE-mediated mechanism may contribute to inflammatory reactions in the skin. OBJECTIVE: This study investigates IgE-sensitization of patients infected with human immunodeficiency virus to a panel of fungal extracts of Candida albicans, Fusarium moniliforme, Penicillium notatum, Pityrosporum ovale, and Trichophyton rubrum. METHODS: Fifteen HIV-positive patients with eczematous skin manifestations and five non atopic healthy controls were evaluated by basophil histamine release and skin prick test with fungal extracts. The extracts were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis under reducing conditions and analyzed by IgE-immunoblotting with sera from the patients and controls. RESULTS: Thirteen of 15 patients (87%) released histamine to one or more of the fungi. Skin prick test was positive to one or more fungi in 7 (47%) patients. Patient sera revealed binding to a wide range of IgE-binding components present in the fungal extracts. The IgE response was most often directed against a 46-kD main protein in the Candida albicans extract. There was no correlation between total serum IgE, CD4+ cell counts, and frequency of IgE-sensitization to fungi. CONCLUSION: The human IgE response in HIV-infected patients appears to be polyspecific and may be directed against various fungi of which Candida albicans may be an important allergen. It is possible that the sensitization is due to frequent infections with Candida albicans in this patient population. No unspecific fungal reactions were noted among control patients. These results suggest that allergen-specific IgE-mediated mechanism may contribute to the pathogenesis of the eczematous skin reaction in HIV-infected patients. PMID- 10480591 TI - Low IgG3 and high IgG4 subclass levels in children with advanced human immunodeficiency virus-type 1 infection and elevated IgE levels. AB - BACKGROUND: IgG3 and IgG4 levels are not always changed in children perinatally infected with human immunodeficiency virus-type 1 (HIV-1). Elevated IgE levels hallmark the TH1 to TH2 switch occurring in advanced infection and such an unbalanced cytokine network may affect the IgG subclass production. OBJECTIVE: To examine the different behaviour of IgG3 and IgG4 in the light of elevated IgE levels. METHODS: IgE and IgG subclass levels were cross-sectionally determined (by radioimmunoassay and enzyme-linked immunosorbent assay, respectively) in 54 HIV-1 perinatally infected children. IgE levels beyond the upper 95% confidence limits (95%CL) of the age-related reference values defined elevated IgE levels. Since immunoglobulin levels physiologically vary with age, individual z-scores of isotype levels were calculated using the upper 95%CL of age-related reference values. RESULTS: Fifteen (27.7%) children had elevated IgE levels. They had lower IgG3 (mean +/- standard deviation: -1.4+/-0.9 versus 6.9+/-0.9; P < .0001) and higher IgG4 (3.1+/-0.6 versus 0.2+/-0.3; P < .0001) z-scores compared with children without elevated IgE levels. Similar IgG1 (11.7+/-1.8 versus 12.6+/-1.3) and IgG2 (-0.22+/-0.8 versus -0.19+/-0.6) z-scores were found. In children with elevated IgE levels, IgE and IgG3 z-scores inversely correlated (r = -0.867; P < .0001), IgE and IgG4 z-scores directly correlated (r = 0.831; P < .0001) and IgG3 and IgG4 z-scores inversely correlated (r = -0.745; P < .001). CONCLUSION: Low IgG3 and high IgG4 levels may be present in HIV-1 advanced disease with elevated IgE levels. Changes may be in line with the TH1 to TH2 switch and contribute to disease progression. PMID- 10480592 TI - Bupropion hydrochloride induced serum sickness-like reaction. AB - BACKGROUND: Bupropion hydrochloride is used frequently in the treatment of depression and in smoking cessation therapy. METHODS: We report a case of bupropion hydrochloride induced serum sickness-like reaction. RESULTS: There was complete resolution of symptoms on discontinuing bupropion and instituting therapy with prednisone and antihistamines. CONCLUSION: This case is seemingly the first report of a serum sickness-like reaction to bupropion hydrochloride. PMID- 10480593 TI - Incidence of cockroach allergy in atopic Italian children. AB - BACKGROUND: Recent studies suggest that sensitization to cockroach allergens is common in urban areas and highlight the importance of cockroach allergens as a risk factor for asthma. OBJECTIVE: To elucidate the prevalence of cockroach allergy in atopic Italian children and to evaluate the correlation between cockroach allergens and other major indoor allergens. METHODS: Four hundred eighty two children with at least one positive skin test response to a common allergen and with personal history of atopic diseases, were investigated. A questionnaire was administered in order to obtain data on epidemiologic and clinical characteristics of the population. Skin tests to 24 common allergens (including extracts of two cockroach species) were carried out and serum total and cockroach specific IgE were evaluated. RESULTS: 61/482 (12.7%) children had positive skin test to cockroach extracts. Among them the frequency of children living in urban areas was higher (P < .05). In addition, a significant correlation between results of skin tests to cockroach and mite allergens was observed (P < .009). No correlation between cockroach allergy and the clinical characteristics of the study population was found. CONCLUSIONS: The results show that cockroach allergy affects a small portion of atopic Italian children and suggest that it is related with living in urban areas and with sensitization to other indoor allergens, such as dust mite. PMID- 10480594 TI - Onset of type 1 diabetes: a dynamical instability. AB - Type 1 diabetes is a disease characterized by progressive loss of beta-cell function due to an autoimmune reaction affecting the islets of Langerhans. It is now generally accepted that cytokines are implicated in the pathogenesis of autoimmune diseases. Animal studies have shown that interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma affect type 1 diabetes development profoundly. It has been suggested that beta-cells are destroyed by cytokine induced free radical formation before cytotoxic T-helper (Th)-lymphocytes and/or autoantibody-mediated cytolysis. This hypothesis is known as the "Copenhagen model." We introduce a mathematical model encompassing the various processes within this framework. The model is expressed in rate equations describing the changes in numbers of beta-cells, macrophages, and Th-lymphocytes. Being concerned with the earliest events, we explore the conditions necessary to maintain self-sustained beta-cell elimination based on the feedback between immune cells and insulin-producing cells. The motivation for this type of analysis becomes clear when we consider the multifactorial and complicated nature of the disease. Indeed, recent research has provided detailed information about the different factors that contribute to the development of the disease, stressing the importance of incorporating these findings into a more general picture. A mathematical formalism allows for a more comprehensive description of the biological problem and can reveal nonintuitive properties of the dynamics. Despite the rather complicated structure of the equations, our main conclusion is simple: onset of type 1 diabetes is due to a collective, dynamical instability, rather than being caused by a single etiological factor. PMID- 10480595 TI - Identification of the docked granule pool responsible for the first phase of glucose-stimulated insulin secretion. AB - The mechanisms underlying the first phase of glucose-stimulated insulin release, the deterioration of which marks the early stages of both type 1 and type 2 diabetes, are essentially unknown. Among many hypotheses, one holds that the first phase is due to a readily releasable pool of insulin-containing granules. We used current knowledge of the mechanisms of exocytosis and the proteins involved in docking granules at the plasma membrane to test this hypothesis. A docked pool of readily releasable granules was identified by immunoprecipitation of the plasma membrane protein syntaxin with a specific antibody and by co immunoprecipitation of soluble N-ethylmaleimide-sensitive factor attachment protein-25 (SNAP-25) and the granule proteins synaptobrevin and synaptotagmin. The four SNARE proteins co-immunoprecipitated each other, thus identifying the core complex associated with docked granules. Using co-immunoprecipitation as a marker for docked granules, we found that the docked pool was rapidly discharged during the first phase of glucose-stimulated insulin release and refilled during the second phase. Other secretagogues also released the pool, whereas the physiological inhibitor norepinephrine blocked its release. Further studies on the nature of this pool of granules should shed light on the causes of its deterioration in the early stages of diabetes and the reasons for deficient insulin release. PMID- 10480596 TI - Long-term regulation of lipolysis and hormone-sensitive lipase by insulin and glucose. AB - Adipocyte glucose transport can be impaired by prolonged hyperglycemic conditions. However, at the whole body level, lipolysis is quantitatively a more important function of adipocytes than glucose uptake. We have therefore investigated the effect of prolonged high glucose and insulin on adipocyte lipolysis in basal conditions or with maximal concentrations of adenosine deaminase (ADA), dibutyryl cyclic-AMP (dbcAMP), or isoproterenol (ISO). Neither insulin nor glucose alone affected basal or maximally stimulated lipolysis. However, insulin plus glucose increased the rate of ADA-, dbcAMP-, and ISO stimulated lipolysis by 40-65%, and the effect was maximal by 8 h. When insulin was kept constant, the half-maximally effective concentration (EC50) of glucose was approximately 2.5 mmol/l. We also demonstrated that the effect is not glutamine-dependent and does not induce insulin resistance of lipolysis. Because the effect of insulin and glucose was evident whether lipolysis was stimulated by ADA, dbcAMP, or ISO, we hypothesized that the expression of the rate-limiting enzyme for lipolysis, hormone-sensitive lipase (HSL), was increased. Our results show that insulin plus glucose-treated cells contain approximately 40% more HSL protein than control cells, in good agreement with the increase in maximally stimulated lipolysis. We conclude that hyperglycemic-hyperinsulinemic conditions increase basal and maximal adipocyte lipolysis by a mechanism that is not glutamine-dependent and involves maintenance of cellular concentrations of HSL. The results also provide evidence that factors other than translocation of HSL to the lipid droplet are necessary to activate the enzyme. PMID- 10480597 TI - Structural model of human glucokinase in complex with glucose and ATP: implications for the mutants that cause hypo- and hyperglycemia. AB - Mutations in human glucokinase are implicated in the development of diabetes and hypoglycemia. Human glucokinase shares 54% identical amino acid residues with human brain hexokinase I. This similarity was used to model the structure of glucokinase by analogy to the crystal structure of brain hexokinase. Glucokinase was modeled with both its substrates, glucose and MgATP, to understand the effect of mutations. The glucose is predicted to form hydrogen bond interactions with the side chains of glucokinase residues Thr 168, Lys 169, Asn 204, Asp 205, Asn 231, and Glu 290, similar to those observed for brain hexokinase I. The magnesium ion is coordinated by the carboxylates of Asp 78 and Asp 205 and the gamma phosphate of ATP. ATP is predicted to form hydrogen bond interactions with residues Gly 81, Thr 82, Asn 83, Arg 85, Lys 169, Thr 228, Lys 296, Thr 332, and Ser 336. Mutations of residues close to the predicted ATP binding site produced dramatic changes in the Km for ATP, the catalytic rate, and a loss of cooperativity, which confirmed our model. Mutations of residues in the glucose binding site dramatically reduced the catalytic activity, as did a mutation that was predicted to disrupt an alpha-helix. Other mutations located far from the active site gave smaller changes in kinetic parameters. In the absence of a crystal structure for glucokinase, our models help rationalize the potential effects of mutations in diabetes and hypoglycemia, and the models may also facilitate the discovery of pharmacological glucokinase activators and inhibitors. PMID- 10480598 TI - Role of the sympathetic nervous system and insulin in enhancing glucose uptake in peripheral tissues after intrahypothalamic injection of leptin in rats. AB - Our previous study demonstrated that microinjection of leptin into the ventromedial hypothalamus (VMH) dramatically increased glucose uptake in the heart, brown adipose tissue (BAT), and skeletal muscles, but not in white adipose tissue (WAT) in conscious unrestrained rats, as assessed in vivo by the 2 [3H]deoxyglucose method. Here we examined the role of the sympathetic nervous system and insulin in enhanced glucose uptake by tissues after hypothalamic leptin injection. Pretreatment with guanethidine significantly suppressed the increased glucose uptake by the tissues in response to leptin injected into the VMH, whereas bilateral adrenal demedullation had no significant effect. Treatment with propranolol but not phenoxybenzamine also decreased significantly enhanced glucose uptake by the tissues. We further examined the interaction of the effects of hypothalamic leptin and insulin administered peripherally by clamping the glucose concentrations at a constant level. When leptin was injected into the VMH and a maximal dose of insulin was administered intravenously, the rates of glucose uptake by the heart, BAT, and skeletal muscles, but not by WAT, markedly increased beyond the values reached by insulin stimulation alone. Surgical sympathetic denervation of BAT abolished the enhancement of glucose uptake in this tissue, decreasing to the level stimulated by insulin alone. These results appear to indicate that leptin in the hypothalamus enhances glucose uptake in certain peripheral tissues through mediation of a beta-adrenergic mechanism for the sympathetic nerves innervating the tissues and that central leptin and peripheral insulin have a synergistic role in augmenting tissue glucose uptake. PMID- 10480599 TI - Neonatal porcine islet cells induce human CD4+, but not CD8+, lymphocyte proliferation and resist cell-mediated cytolytic injury in vitro. AB - Xenotransplantation of porcine tissue to human recipients promises to alleviate the organ shortage. Human antibody-mediated and cell-mediated immune responses against porcine grafts, however, represent barriers to successful xenotransplantation. We compared neonatal porcine islet cells (NPICs) and neonatal porcine splenocytes for the ability to stimulate proliferation of human peripheral blood lymphocytes (PBLs), and for their susceptibility to human natural killer (NK) and cytotoxic T-lymphocyte (CTL)-mediated lysis. Human peripheral blood CD4+ lymphocytes showed strong proliferation in response to NPICs, likely because of occasional swine leukocyte antigen (SLA) class II+ cells in the NPIC preparations. In contrast, human peripheral blood CD8+ lymphocytes did not proliferate in response to NPICs, although they showed clear responses to both porcine splenocytes and endothelial cells. Both human CTL-raised-against porcine splenocytes and endogenous NK cells lysed porcine splenocytes, but the same cells showed little or no lytic activity against NPICs. Lysis of porcine splenocyte targets was completely abrogated by pretreatment of the human NK or CTL populations with concana-mycin A, suggesting a perforin-dependent effector mechanism. Pretreatment of the NPIC targets with proinflammatory porcine cytokines to upregulate SLA class I expression failed to enhance human CTL mediated lysis. However, lysis of NPICs by human CTLs could be elicited when a lectin was added to form stable effector:target cell conjugates. It appears that NPICs do not express sufficiently high levels of co-stimulatory and/or adhesion molecules to either activate human CD8+ T-cells or to be effective targets for activated human CTLs. These data suggest that NPICs may not be destroyed by NK- or CTL-mediated lytic mechanisms after transplantation into humans. PMID- 10480600 TI - Insulin B-chain reactive CD4+ regulatory T-cells induced by oral insulin treatment protect from type 1 diabetes by blocking the cytokine secretion and pancreatic infiltration of diabetogenic effector T-cells. AB - The mechanism of protection from type 1 diabetes conferred by regulatory T-cells induced by oral insulin treatment of NOD mice is not well understood. We demonstrate that oral insulin feeding of NOD mice induces the function of insulin B-chain reactive CD4+ regulatory T-cells, which compete with diabetogenic effector T-cells for the recognition of insulin in NOD.Scid recipient mice. These effector T-cells become deprived of interleukin (IL)-2 and interferon (IFN)-gamma and are unable to expand and migrate to the pancreas. Type 1 diabetes-protective splenic regulatory T-cells secrete relatively little transforming growth factor (TGF)-beta1, suggesting that TGF-beta may not contribute to the inactivation of effector T-cells in NOD.Scid recipients. The observed preferential infiltration of insulin-reactive regulatory T-cells rather than effector T-cells in the pancreas results in a nondestructive insulitis that correlates with an increased intrapancreatic expression of macrophage inflammatory protein-1beta. Thus, oral insulin therapy overcomes a deficiency in regulatory T-cells and protects against type 1 diabetes by inducing insulin B-chain reactive regulatory T-cells to block cytokine secretion and migration of diabetogenic effector T-cells to the pancreas. Our data emphasize that continuous oral insulin feeding over a prolonged period is required to prevent type 1 diabetes. PMID- 10480601 TI - Adenoviral gene transfer of the interleukin-1 receptor antagonist protein to human islets prevents IL-1beta-induced beta-cell impairment and activation of islet cell apoptosis in vitro. AB - The beta-cells in the pancreatic islets of Langerhans are the targets of autoreactive T-cells and are destroyed in type 1 diabetes. Macrophage-derived interleukin-1beta (IL-1beta) is important in eliciting beta-cell dysfunction and initiating beta-cell damage in response to microenvironmental changes within islets. In particular, IL-1beta can impair glucose-stimulated insulin production in beta-cells in vitro and can sensitize them to Fas (CD95)/FasL-triggered apoptosis. In this report, we have examined the ability to block the detrimental effects of IL-1beta by genetically modifying islets by adenoviral gene transfer to express the IL-1 receptor antagonist protein. We demonstrate that adenoviral gene delivery of the cDNA encoding the interleukin-1 receptor antagonist protein (IL-1Ra) to cultured islets results in protection of human islets in vitro against IL-1beta-induced nitric oxide formation, impairment in glucose-stimulated insulin production, and Fas-triggered apoptosis activation. Our results further support the hypothesis that IL-1beta antagonism in in situ may prevent intra islet proinsulitic inflammatory events and may allow for an in vivo gene therapy strategy to prevent insulitis and the consequent pathogenesis of diabetes. PMID- 10480602 TI - Normoglycemia and preserved insulin secretory reserve in diabetic patients 10-18 years after pancreas transplantation. AB - Pancreas transplantation is a controversial form of therapy for type I diabetes. A major obstacle to acceptance of this procedure for many physicians is the lack of demonstrable long-term success. We performed these studies to assess the hypothesis that successful pancreas transplantation is efficacious in normalizing endogenous insulin secretion and glycemia in the long term (1-2 decades). Sixteen patients with a history of diabetic complications who had undergone a transplant 10-18 years earlier involving either a whole or a segment of pancreas were recruited for measurements of fasting plasma glucose, HbA1c, intravenous glucose tolerance, and insulin secretory reserve. All patients were taking immunosuppressive drugs, but none was using insulin or other hypoglycemic agents. All recipients had normal levels of fasting blood glucose, intravenous glucose tolerance, and HbA1c, and 15 of 16 stated that their quality of life had improved after transplantation. They had intact acute insulin responses to intravenous pulses of glucose and to arginine and insulin secretory reserve. Glucose potentiation of arginine-induced insulin secretion, the measure of insulin secretory reserve, correlated significantly (r = 0.095, P < 0.001) with the acute insulin response to intravenous glucose, rendering the latter a much simpler and valid measure of functional beta-cell mass. We conclude that successful pancreas transplants are efficacious for periods as long as 1-2 decades in returning euglycemia to type 1 diabetic patients by restoring endogenous insulin secretion and insulin secretory reserve. Thus, concern about long-term deterioration, as distinct from rejection, should not be a major obstacle when deciding whether to recommend pancreas transplantation. PMID- 10480603 TI - Corticotropin-releasing factor modulation of Ca2+ influx in rat pancreatic beta cells. AB - The effects of corticotropin-releasing factor (CRF) on the intracellular concentration of Ca2+ were studied in isolated single beta-cells of the rat islet. Immunohistochemical staining using CRF-receptor antibodies revealed the presence of both type 1 (CRF-R1) and type 2 (CRF-R2) receptors for CRF in the majority of islet cells. CRF (2 nmol/l) increased cytosolic Ca2+ concentration under 2.8 mmol/l glucose, dependent upon extracellular Ca2+. CRF caused depolarization of the cell membrane, which was followed by action potentials under 2.8 mmol/l glucose. The dose-response relationships of CRF-induced depolarization in the presence of 1 micromol/l nifedipine produced a bell-shaped curve, showing the peak response at 2 nmol/l. In the whole-cell patch-clamp recording, CRF enhanced Ca2+ currents through L-type Ca2+ channels in a dose dependent manner similar to that for depolarization. In cells pretreated with Rp deastereomer of adenosine cyclic 3',5'-phosphorothiolate (100 micromol/l), neither depolarization nor an increase in the Ca2+ current was caused by CRF at concentrations <2 nmol/l. In these cells, CRF at 20 nmol/l reduced the Ca2+ current. These results suggest that in single beta-cells of rat islets, CRF, through its own receptor, potentiates Ca2+ influx through the L-type Ca2+ channel by activation of the cAMP/protein kinase A signaling pathway. CRF at a high concentration also shows an inhibitory effect on the Ca2+ current through an unknown signaling pathway. PMID- 10480604 TI - Glucose-fatty acid cycle to inhibit glucose utilization and oxidation is not operative in fatty acid-cultured islets. AB - The glucose-fatty acid cycle of Randle entails two elements: decreased pyruvate dehydrogenase (PDH) activity, which inhibits glucose oxidation, and inhibition of phosphofructokinase (PFK) by a rise in citrate so that glucose-6-phosphate (G-6 P) levels increase, thereby inhibiting hexokinase activity and hence glucose utilization. Chronic exposure of islets to long-chain fatty acids (FA) is reported to lower PDH activity, but the effect on glucose oxidation and glucose induced insulin secretion is uncertain. We investigated rat islets that were cultured for 4 days with 0.25 mmol/l oleate/5.5 mmol/l glucose. Glucose oxidation was doubled at 2.8 mmol/l glucose and unchanged at 27.7 mmol/l glucose in the FA cultured islets despite a 35% decrease in assayed PDH activity. Pyruvate content was increased 60%, which may well compensate for the decreased PDH activity and maintain flux through the citric acid cycle. However, a greater diversion of pyruvate metabolism through the pyruvate-malate shuttle is suggested by unchanged pyruvate carboxylase Vmax and a fourfold higher release of malate from isolated mitochondria. The FA-cultured islets also showed increased basal glucose usage and insulin secretion together with a lowered level of G-6-P and 50% reductions in citrate synthase Vmax and the citrate content. Thus, the effects of chronic FA exposure on islet glucose metabolism differ from the glucose-fatty acid interactions reported in some other tissues. PMID- 10480605 TI - A defect late in stimulus-secretion coupling impairs insulin secretion in Goto Kakizaki diabetic rats. AB - A widely accepted genetically determined rodent model for human type 2 diabetes is the Goto-Kakizaki (GK) rat; however, the lesion(s) in the pancreatic islets of these rats has not been identified. Herein, intact islets from GK rats (aged 8-14 weeks) were studied, both immediately after isolation and after 18 h in tissue culture. Despite intact contents of insulin and protein, GK islets had markedly deficient insulin release in response to glucose, as well as to pure mitochondrial fuels or a non-nutrient membrane-depolarizing stimulus (40 mmol/l K+). In contrast, mastoparan (which activates GTP-binding proteins [GBPs]) completely circumvented any secretory defect. Basal and stimulated levels of adenine and guanine nucleotides, the activation of phospholipase C by Ca2+ or glucose, the secretory response to pertussis toxin, and the activation of selected low-molecular weight GBPs were not impaired. Defects were found, however, in the autophosphorylation and catalytic activity of cytosolic nucleoside diphosphokinase (NDPK), which may provide compartmentalized GTP pools to activate G-proteins; a deficient content of phosphoinositides was also detected. These studies identify novel, heretofore unappreciated, defects late in signal transduction in the islets of our colony of GK rats, possibly occurring at the site of activation by NDPK of a mastoparan-sensitive G-protein-dependent step in exocytosis. PMID- 10480606 TI - Hypothalamic glucose sensor: similarities to and differences from pancreatic beta cell mechanisms. AB - Glucose-responsive neurons in the ventromedial hypothalamus (VMH) are stimulated when glucose increases from 5 to 20 mmol/l and are thought to play an essential role in regulating metabolism. The present studies examined the role of glucose metabolism in the mechanism by which glucose-responsive neurons sense glucose. The pancreatic, but not hepatic, form of glucokinase was expressed in the VMH, but not cerebral cortex, of adult rats. In brain slice preparations, the transition from 5 to 20 mmol/l glucose stimulated approximately 17% of the neurons (as determined by single-cell extracellular recording) from VMH but none in cortex. In contrast, most cells in both VMH and cortex were silent below 1 mmol/l and active at 5 mmol/l glucose. Glucosamine, 2-deoxyglucose, phloridzin, and iodoacetic acid blocked the activation of glucose-responsive neurons by the transition from 5 to 20 mmol/l glucose. Adding 15 mmol/l mannose, galactose, glyceraldehyde, glycerol, and lactate to 5 mmol/l glucose stimulated glucose responsive neurons. In contrast, adding 15 mmol/l pyruvate to 5 mmol/l glucose failed to activate glucose-responsive neurons, although pyruvate added to 0 mmol/l glucose permitted neurons to maintain activity. Tolbutamide activated glucose-responsive neurons; however, diazoxide only blocked the effect of glucose in a minority of neurons. These data suggest that glucose-responsive neurons sense glucose through glycolysis using a mechanism similar to the mechanism of pancreatic beta-cells, except that glucose-responsive neurons are stimulated by glycerol and lactate, and diazoxide does not generally block the effect of glucose. PMID- 10480607 TI - Expression of alpha-1 proteinase inhibitor in human islet microvascular endothelial cells. AB - There is a microcirculation system within the islets of Langerhans. However, little is known about the phenotypic and functional characterization of islet microvascular endothelial cells (MVEC). In this study, we purified MVEC from human pancreatic islets by using Ulex europaeus (Sigma, St. Louis, MO) agglutinin 1 (UEA-1)-coated dynabeads (Dynal A.S., Oslo, Norway). These purified human islet MVEC (HI-MVEC) express von Willebrand factor, take up high levels of acetylated LDL, and upregulate endothelial cell leukocyte adhesion molecule 1 in response to tumor necrosis factor-alpha. Ultrastructure examination shows the presence of microvilli and fenestrations on the cell surface, Weibel-Palade bodies in the cytoplasm, and tight junctions between cells. Furthermore, we show that vascular endothelial cell growth factor contributes to the formation of surface fenestrations on cultured HI-MVEC. After purification, HI-MVEC exhibit a very low proliferation capacity and are strongly resistant to trypsin, compared with other original MVEC. We also demonstrate that alpha-1 proteinase inhibitor (Api) is expressed on HI-MVEC and specifically located at the area of cell-cell junctions. By reverse transcription-polymerase chain reaction, a significant messenger RNA band of Api was found only in HI-MVEC, but not in other organ-derived MVEC, indicating that expression of Api is islet MVEC specific. Antibodies to Api significantly reversed the resistance to trypsin and promoted proliferation of HI MVEC, suggesting that these specific functional characteristics of HI-MVEC are related to the expression of Api. These results indicate that HI-MVEC exhibit some specific morphological and functional characteristics that differ from MVEC derived from other organs. PMID- 10480608 TI - Comparison of tests of beta-cell function across a range of glucose tolerance from normal to diabetes. AB - Adequate comparisons of the relative performance of different tests of beta-cell function are not available. We compared discrimination of commonly used in vivo tests of beta-cell function across a range of glucose tolerance in seven subjects with normal glucose tolerance (NGT), eight subjects with impaired glucose tolerance (IGT), and nine subjects with type 2 diabetes. In random order, each subject underwent two of each of the following tests: 1) frequently sampled 0.3 g/kg intravenous glucose tolerance test (FSIVGTT) with MinMod analysis; 2) homeostasis model assessment (HOMA) from three samples at 5-min intervals with a model incorporating immunoreactive or specific insulin measurements; and 3) continuous infusion of 180 mg x min(-1) x m(-2) glucose with model assessment (CIGMA) of three samples at 50, 55, and 60 min (1-h CIGMA) and at 110, 115, and 120 min (2-h CIGMA). The discrimination of each test was assessed by the ratio of the within-subject SD to the underlying between-subject SD, the discriminant ratio (DR). The degree to which tests measured the same physiological variable was assessed using Pearson's correlation coefficient adjusted for attenuation due to test imprecision. An unbiased line of equivalence, taking into account the imprecision of both tests, was used to compare results. Beta-cell function assessed from HOMA and beta-cell function assessed from CIGMA (CIGMA%beta) (using immunoreactive insulin) had higher DRs than first-phase intravenous glucose tolerance test-derived incremental insulin peak, area, insulin-to-glucose index, and acute insulin response to glucose from FSIVGTT-MinMod. CIGMA%beta (immunoreactive insulin) had the highest DR. FSIVGTT-derived first-phase insulin response tests correlated only moderately with HOMA and CIGMA. Using specific rather than immunoreactive insulin for HOMA and CIGMA did not improve discriminatory power. Simple tests such as HOMA and CIGMA, using immunoreactive insulin, offer better beta-cell function discrimination across subjects with NGT, IGT, and type 2 diabetes than measurements derived from FSIVGTT first-phase insulin response. PMID- 10480609 TI - Sympathetic activation of leptin via the ventromedial hypothalamus: leptin induced increase in catecholamine secretion. AB - Leptin is an adipocyte-derived blood-borne satiety factor that acts directly on the hypothalamus, thereby regulating food intake and energy expenditure. We have demonstrated that the hypothalamic arcuate nucleus (Arc) is a primary site of the satiety effect of leptin (Neurosci Lett 224:149-152, 1997). To explore the hypothalamic pathway of sympathetic activation of leptin, we examined the effects of a single intravenous or intracerebroventricular injection of recombinant human leptin on catecholamine secretion in rats. We also examined the effects of direct microinjection of leptin into the ventromedial hypothalamus (VMH), Arc, paraventricular nucleus (PVN), and dorsomedial hypothalamus (DMH) in rats. To further assess whether sympathetic activation of leptin is mediated via the VMH, we also examined the effects of a single intravenous injection of leptin in VMH lesioned rats. A single injection of leptin (0.25-1.0 mg i.v./rat or 0.5-2.0 pg i.c.v./rat) increased plasma norepinephrine (NE) and epinephrine (EPI) concentrations in a dose-dependent manner. Plasma NE and EPI concentrations were increased significantly when leptin was injected directly into the VMH but were unchanged when injected into the Arc, PVN, and DMH. Plasma NE and EPI concentrations were unchanged in VMH-lesioned rats that received a single intravenous injection of leptin. The present study provides evidence that a leptin-induced increase in catecholamine secretion is mediated primarily via the VMH and suggests the presence of distinct hypothalamic pathways mediating the satiety effect and sympathetic activation of leptin. PMID- 10480610 TI - T-1095, an inhibitor of renal Na+-glucose cotransporters, may provide a novel approach to treating diabetes. AB - T-1095A and T-1095 are synthetic agents derived from phlorizin, a specific inhibitor of Na+-glucose cotransporters (SGLTs). Unlike phlorizin, T-1095 is absorbed into the circulation via oral administration, is metabolized to the active form, T-1095A, and suppresses the activity of SGLTs in the kidney. Orally administered T-1095 increases urinary glucose excretion in diabetic animals, thereby decreasing blood glucose levels. Indeed, the postprandial hyperglycemia after a meal load was shown to be suppressed by this compound in streptozotocin (STZ)-induced diabetic rats. With long-term T-1095 treatment, both blood glucose and HbA1c levels were reduced in STZ-induced diabetic rats and yellow KK mice. In addition, there was amelioration of abnormal carbohydrate metabolism, i.e., hyperinsulinemia and hypertriglyceridemia, and of the development of microalbuminuria, in yellow KK mice. Thus, T-1095 may be a useful antidiabetic drug, providing a novel therapeutic approach for diabetes. PMID- 10480611 TI - Altered hypothalamic function in response to glucose ingestion in obese humans. AB - The hypothalamus plays a central role in the regulation of energy intake and feeding behavior. However, the presence of a functional abnormality in the hypothalamus in humans that may be related to excess energy intake and obesity has yet to be demonstrated in vivo. We, therefore, used functional magnetic resonance imaging (fMRI) to monitor hypothalamic function after oral glucose intake. The 10 obese (34 +/- 2 years of age, BMI 34.2 +/- 1.3 kg/m2) and 10 lean (32 +/- 4 years of age, BMI 22.0 +/- 0.9 kg/m2) subjects with normal glucose tolerance ingested 75 g of glucose while a midsagittal slice through the hypothalamus was continuously imaged for 50 min using a conventional T2*-weighted gradient-echo pulse sequence. After glucose ingestion, lean subjects demonstrated an inhibition of the fMRI signal in the areas corresponding to the paraventricular and ventromedial nuclei. In obese subjects, this inhibitory response was markedly attenuated (4.8 +/- 1.3 vs. 7.0 +/- 0.6% inhibition, P < 0.05) and delayed (9.4 +/- 0.5 vs. 6.4 +/- 0.5 min, P < 0.05) compared with that observed in lean subjects. The time taken to reach the maximum inhibitory response correlated with the fasting plasma glucose (r = 0.75, P < 0.001) and insulin (r = 0.47, P < 0.05) concentrations in both lean and obese subjects. These results demonstrate in vivo, for the first time, the existence of differential hypothalamic function in lean and obese humans that may be secondary to obesity. PMID- 10480612 TI - Impaired glucose transport and insulin receptor tyrosine phosphorylation in skeletal muscle from obese women with gestational diabetes. AB - Women who develop gestational diabetes mellitus (GDM) have severe insulin resistance and markedly increased risk to develop subsequent type 2 diabetes. We investigated the effects of pregnancy and GDM on glucose transport activity and the expression and phosphorylation of the insulin receptor and insulin receptor substrate (IRS)-1 in human skeletal muscle fiber strips in vitro. Rectus abdominis muscle biopsies were obtained at the time of cesarean section from 11 pregnant women with normal glucose tolerance (pregnant control), 7 pregnant women with GDM, and 11 nonpregnant women undergoing elective surgery (nonpregnant control). Subjects were matched for age and similar degree of obesity. The rate of maximal insulin (10(-7) mol/l)-stimulated 2-deoxyglucose transport was reduced by 32% (P < 0.05) in muscle strips from the pregnant control group and even further in GDM subjects by 54% (P < 0.05 vs. pregnant control). The maximal effect of insulin on tyrosine phosphorylation of the insulin receptor was 37% lower (P < 0.05) in GDM subjects than in pregnant control subjects and was not related to changes in the abundance of the insulin receptor. Compared with nonpregnant control subjects, maximal insulin-stimulated IRS-1 tyrosine phosphorylation was significantly lower by 59 +/- 24% (mean +/- SD) (P < 0.05) and 62 +/- 28% (P < 0.05) in pregnant control and GDM subjects, respectively. This was reflected by a 23% (P < 0.05) and 44% (P < 0.002) reduction in IRS-1 protein levels in muscle from pregnant control and GDM subjects. Both pregnant control and GDM subjects exhibited a 1.5- to 2-fold increase in the levels of IRS 2 (P < 0.01) and p85alpha regulatory subunit of phosphatidylinositol (PI) 3 kinase (P < 0.05), despite reduced glucose transport activity. These data indicate that insulin resistance to glucose transport during pregnancy is uniquely associated with a decrease in IRS-1 tyrosine phosphorylation, primarily due to decreased expression of IRS-1 protein. However, in GDM subjects, a decrease in tyrosine phosphorylation of the insulin receptor beta-subunit is associated with further decreases in glucose transport activity. Thus, impaired insulin receptor autophosphorylation is an important early distinction underlying muscle insulin resistance in young women with GDM, and it may underlie future risk for the development of type 2 diabetes. PMID- 10480613 TI - Nitric oxide synthase inhibition reduces leg glucose uptake but not blood flow during dynamic exercise in humans. AB - Nitric oxide (NO) appears to play a role in contraction-stimulated glucose uptake in isolated rodent skeletal muscle; however, no studies have examined this question in humans. Seven healthy men completed two 30-min bouts of supine cycling exercise at 60 +/- 2% peak pulmonary oxygen uptake (VO2 peak), separated by 90 min of rest. The NO synthase inhibitor N(G)-monomethyl-L-arginine ([L NMMA]; total dose 5 mg/kg body weight) or saline (control) were administered via the femoral artery for the final 20 min of exercise in a randomized blinded crossover design. L-Arginine (5 mg/kg body weight) was co-infused during the final 5 min of each exercise bout. Leg blood flow (LBF) was measured by thermodilution in the femoral vein, and leg glucose uptake was calculated as the product of LBF and femoral arteriovenous (AV) glucose difference. L-NMMA infusion significantly (P < 0.05) reduced leg glucose uptake compared with control (48 +/- 12% lower at 15 min, mean +/- SE). The reduction in glucose uptake was due solely to a decrease in AV glucose difference, as there was no effect of L-NMMA infusion on LBF during exercise. Co-infusion of L-arginine restored glucose uptake during L-NMMA infusion to levels similar to control. These results indicate that NO production contributes substantially to exercise-mediated skeletal muscle glucose uptake in humans independent of skeletal muscle blood flow. PMID- 10480614 TI - Increased glucose metabolism and insulin sensitivity in transgenic skinny mice overexpressing leptin. AB - Excess of body fat, or obesity, is a major health problem and confers a higher risk of cardiovascular and metabolic disorders such as diabetes, hypertension, and coronary heart disease. Leptin is an adipocyte-derived satiety factor that plays an important role in the regulation of energy homeostasis, and its synthesis and secretion are markedly increased in obese subjects. To explore the metabolic consequences of an increased amount of leptin on a long-term basis in vivo, we generated transgenic skinny mice with elevated plasma leptin concentrations comparable to those in obese subjects. Overexpression of leptin in the liver has resulted in complete disappearance of white and brown adipose tissue for a long period of time in mice. Transgenic skinny mice exhibit increased glucose metabolism accompanied by the activation of insulin signaling in the skeletal muscle and liver. They also show small-sized livers with a marked decrease in glycogen and lipid storage. The phenotypes are in striking contrast to those of recently reported animal models of lipoatrophic diabetes and patients with lipoatrophic diabetes with reduced amount of leptin. The present study provides evidence that leptin is an adipocyte-derived antidiabetic hormone in vivo and suggests its pathophysiologic and therapeutic implications in diabetes. PMID- 10480615 TI - Improvement in insulin resistance and the restoration of reduced phosphodiesterase 3B gene expression by pioglitazone in adipose tissue of obese diabetic KKAy mice. AB - Phosphodiesterase (PDE) 3B is a key enzyme in the mediation of the antilipolytic action of insulin in adipocytes, and activation of this molecule results in a reduced output of free fatty acids (FFAs). An elevation of serum FFAs is known to cause insulin resistance in skeletal muscle and liver, which could be the primary cause of type 2 diabetes. To elucidate whether PDE3B is involved in this disease, we examined the PDE3B gene expression in epididymal fat tissues of obese insulin resistant diabetic KKAy mice. We also examined the effect of an insulin sensitizing drug, pioglitazone, on this gene expression. In adipose tissue of KKAy mice, PDE3B mRNA and its corresponding protein were reduced to 48 and 43% of those in C57BL/6J control mice. Basal and insulin-stimulated membrane-bound PDE activities were also decreased to 50 and 36% of those in the controls, respectively. Pioglitazone increased both PDE3B mRNA and protein levels by 1.8 fold of those in untreated KKAy mice. Basal and insulin-induced membrane-bound PDE activities were also increased by 1.6- and 2.0-fold, respectively. Pioglitazone reduced the elevated levels of serum insulin, glucose, FFAs, and triglyceride in KKAy mice. Thus, the reduced PDE3B gene expression in adipose tissues could be the primary event in the development of insulin resistance in KKAy mice, which was improved by pioglitazone possibly because of the restoration of the reduced PDE3B gene expression. PMID- 10480616 TI - Overnight lowering of free fatty acids with Acipimox improves insulin resistance and glucose tolerance in obese diabetic and nondiabetic subjects. AB - Obesity is commonly associated with elevated plasma free fatty acid (FFA) levels, as well as with insulin resistance and hyperinsulinemia, two important cardiovascular risk factors. What causes insulin resistance and hyperinsulinemia in obesity remains uncertain. Here, we have tested the hypothesis that FFAs are the link between obesity and insulin resistance/hyperinsulinemia and that, therefore, lowering of chronically elevated plasma FFA levels would improve insulin resistance/hyperinsulinemia and glucose tolerance in obese nondiabetic and diabetic subjects. Acipimox (250 mg), a long-acting antilipolytic drug, or placebo was given overnight (at 7:00 P.M., 1:00 A.M., 7:00 A.M.) to 9 lean control subjects, 13 obese nondiabetic subjects, 10 obese subjects with impaired glucose tolerance, and 11 patients with type 2 diabetes. Euglycemic hyperinsulinemic clamps and oral glucose tolerance tests (75 g) were performed on separate mornings after overnight Acipimox or placebo treatment. In the three obese study groups, Acipimox lowered fasting levels of plasma FFAs (by 60-70%) and plasma insulin (by approximately 50%). Insulin-stimulated glucose uptake during euglycemic-hyperinsulinemic clamping was more than twofold higher after Acipimox than after placebo. Areas under the glucose and insulin curves during oral glucose tolerance testing were both approximately 30% lower after Acipimox administration than after placebo. We conclude that lowering of elevated plasma FFA levels can reduce insulin resistance/hyperinsulinemia and improve oral glucose tolerance in lean and obese nondiabetic subjects and in obese patients with type 2 diabetes. PMID- 10480617 TI - Autoantibody against N(epsilon)-(carboxymethyl)lysine: an advanced glycation end product of the Maillard reaction. AB - Prolonged incubation of proteins with reducing sugar produces advanced glycation end products (AGEs), which are implicated as factors for aging and diabetic complications. We previously demonstrated the presence of N(epsilon) (carboxymethyl)lysine (CML), one of the main AGE structures, in human and animal tissues using a monoclonal anti-CML antibody (6D12). These findings suggest that CML structures present in vivo could serve as immunogens to generate autoantibodies. This suggestion was tested in the present study. First, plasma samples from diabetic rats reacted positively with AGE bovine serum albumin (BSA). These reactivities increased with the duration of diabetic states and were inhibited specifically by CML-BSA. Second, a fraction purified from plasma of diabetic patients, which bound to AGE-BSA, showed a positive reaction to CML-BSA and furthermore also to human lens proteins, which are known to undergo CML modification in vivo. Finally, patients with renal failure caused by diabetes or nondiabetic pathologies had a higher autoantibody activity against CML structure than that in normal subjects or diabetic patients without renal failure. These results indicate that CML accumulated in vivo serves as an immunological epitope to generate an autoantibody specific for CML that might be used as a potential marker for diabetic nephropathy or chronic renal failure. PMID- 10480618 TI - Hyperketonemia can increase lipid peroxidation and lower glutathione levels in human erythrocytes in vitro and in type 1 diabetic patients. AB - Recent studies have suggested that elevated cellular lipid peroxidation may play a role in the development of cellular dysfunction and other complications of diabetes. People with type 1 diabetes frequently encounter elevated levels of the ketone bodies acetoacetate (AA), beta-hydroxybutyrate (BHB), and acetone (ACE). This study was undertaken to test the hypothesis that ketosis might increase lipid peroxidation and lower glutathione (GSH) levels of red blood cells (RBCs) in diabetic patients. This study demonstrates that incubation of AA with normal RBCs in phosphate-buffered saline (37 degrees C for 24 h) resulted in marked GSH depletion, oxidized glutathione accumulation, hydroxyl radical generation, and increased membrane lipid peroxidation. Increases in oxygen radicals and lipid peroxidation and depletion of GSH in RBCs were not observed with BHB or ACE treatments. Similarly, there was a significant generation of superoxide ion radicals even in a cell-free buffer solution of AA, but not in that of BHB. The presence of BHB together with AA did not influence the capacity of AA to generate oxygen radicals in a cell-free solution or the increase in lipid peroxidation of RBCs incubated with AA. The antioxidants vitamin E and N-acetylcysteine (NAC) blocked increase in lipid peroxidation in AA-treated RBCs. To examine the effects of ketone bodies in vivo, studies were performed that showed a significant decrease in GSH and an increase in lipid peroxidation levels in RBCs of hyperketonemic diabetic patients, but not in normoketonemic type 1 diabetic patients, when compared with age-matched normal subjects. This study demonstrates that elevated levels of the ketone body AA can increase lipid peroxidation and lower GSH levels of RBCs in people with type 1 diabetes. PMID- 10480619 TI - Microvascular and macrovascular reactivity is reduced in subjects at risk for type 2 diabetes. AB - Abnormalities in vascular reactivity in the micro- and macrocirculation are well established in type 2 diabetes. However, little is known about changes in vascular reactivity in those at risk for developing type 2 diabetes. To address this situation, the vascular reactivity in both the micro- and macrocirculation was studied in four age and sex comparable groups: 30 healthy normoglycemic subjects with no history of type 2 diabetes in a first-degree relative (controls), 39 healthy normoglycemic subjects with a history of type 2 diabetes in one or both parents (relatives), 32 subjects with impaired glucose tolerance (IGT), and 42 patients with type 2 diabetes without vascular complications (diabetes). Laser Doppler perfusion imaging was used to measure vasodilation in the forearm skin in response to iontophoresis of 1% acetylcholine chloride (Ach) (endothelium-dependent) and 1% sodium nitroprusside (SNP) (endothelium independent), whereas high-resolution ultrasound images were used to measure brachial artery diameter changes during reactive hyperemia. Plasma concentrations of endothelin-1 (ET-1), von Willebrand factor (vWF), soluble intercellular adhesion molecule (sICAM), and soluble vascular cell adhesion molecule (sVCAM) were also measured as indicators of endothelial cell activation. The vasodilatory responses to Ach, expressed as percent increase of blood flow over baseline, were reduced in relatives (98 +/- 48, mean +/- SD), IGT (94 +/- 52), and diabetes (74 +/- 45) compared with controls (126 +/- 67) (P < 0.001 controls versus relatives, IGT, and diabetes). The responses to SNP were similarly reduced: controls (123 +/ 46), relatives (85 +/- 46), IGT (83 +/- 48), and diabetes (65 +/- 31) (P < 0.001 controls versus relatives, IGT, and diabetes) as were the responses in the brachial artery diameter during reactive hyperemia: controls (13.7 +/- 6.1), relatives (10.5 +/- 6.7), IGT (9.8 +/- 4.5), and diabetes (8.4 +/- 5.0) (P < 0.01 controls versus relatives, IGT, and diabetes). Women had greater responses than men in both the micro- and macrovascular circulatory tests, but a similar progressive reduction was observed in both sexes with increasing degrees of glucose intolerance. A significant inverse correlation was found between microvascular reactivity and systolic blood pressure, fasting plasma glucose, HDL cholesterol, fasting plasma insulin, and homeostasis model assessment (HOMA) values, an index of insulin resistance. BMI and diastolic blood pressure had a significant inverse correlation only with endothelium-dependent vasodilation. In the macrocirculation, systolic blood pressure, HbA1c, HDL cholesterol, and HOMA had significant correlation with brachial artery diameter changes. Compared with control subjects, ET-1 was significantly higher in all groups, vWF was higher only in the diabetic group, sICAM levels were higher in the IGT and diabetic groups, while sVCAM concentrations were higher in the relatives and those with diabetes (P < 0.05). On stepwise multivariate analysis, age, sex, fasting plasma glucose, and BMI were the most important contributing factors to the variation of vascular reactivity. Addition of all clinical and biochemical measures explained only 32-37% of the variation in vascular reactivity. These results suggest that abnormalities in vascular reactivity and biochemical markers of endothelial cell activation are present early in individuals at risk of developing type 2 diabetes, even at a stage when normal glucose tolerance exists, and that factors in addition to insulin resistance may be operative. PMID- 10480620 TI - A quantitative trait locus influencing BMI maps to the region of the beta-3 adrenergic receptor. AB - The beta-3 adrenergic receptor (ADRB3) has been implicated as a regulator of energy expenditure, and a polymorphism in codon 64 of this gene (Trp64Arg) has been associated in some studies with obesity and insulin resistance. However, many studies have failed to detect an effect of this variant, and the importance of the Trp64Arg variant in human obesity remains controversial. We performed a quantitative linkage analysis of the ADRB3 and obesity, using 12 markers (including the intragenic Trp64Arg polymorphism) spanning a 57-cM region of chromosome 8. The study population consisted of 470 individuals from 10 large multigenerational families of Mexican-American ancestry residing in San Antonio, TX. In two-point analysis, logarithm of odds (LOD) scores >1.0 were observed for six markers surrounding ADRB3 in a 33-cM region spanned by markers D8S1477 and D8S1136. The multipoint LOD score was 3.21, occurring between markers D8S1121 and ADRB3, approximately 2-3 cM from ADRB3. Adjusting for the presence of the Arg64 allele or excluding from the analysis the 11 individuals homozygous for the Arg64 allele did not reduce the evidence for linkage. A genome scan was conducted at 10 cM map density to detect other loci influencing variation in BMI. Multipoint LOD scores >1.0 were observed in four other regions, including two on chromosome 17, one on chromosome 6q, and one on chromosome 2p. These data suggest that the ADRB3 should continue to be regarded as a strong candidate gene for obesity even though evidence for an effect of the Trp64Arg polymorphism could not be established. It is also possible that a gene closely linked to ADRB3 may influence susceptibility to obesity. PMID- 10480621 TI - Variants of the insulin receptor substrate-1 and fatty acid binding protein 2 genes and the risk of type 2 diabetes, obesity, and hyperinsulinemia in African Americans: the Atherosclerosis Risk in Communities Study. AB - We conducted a community-based case-control study of African-American men and women in the Atherosclerosis Risk in Communities Study. The allele frequencies of the Gly972Arg variant of the insulin receptor substrate-1 (IRS-1) gene and the Ala54Thr variant of the fatty acid binding protein 2 (FABP2) gene were compared in 992 normal control subjects and three patient groups: 1) 321 type 2 diabetic individuals, 2) 260 severely obese individuals, and 3) 258 markedly hyperinsulinemic individuals without diabetes. Allele frequencies of Gly972Arg IRS-1 and Ala54Thr FABP2 were 0.07 and 0.22, respectively; there were no differences in allele or genotype frequencies between patients and control subjects for either gene variant. In weighted linear regression of all patients and control subjects, the presence of the IRS-1 gene variant was associated with a 0.85 (0.42) kg/m2 higher BMI (P = 0.04). In addition, individuals with at least one IRS-1 Arg972 allele and two FABP2 Thr54 alleles had a BMI of 33.3 (7.9) kg/m2, compared with 30.0 (6.3) kg/m2 for those with neither allele (P = 0.05). These results suggest that in African-Americans, these variants in the IRS-1 and FABP2 genes are not associated with the risk of type 2 diabetes, severe obesity, or marked hyperinsulinemia, but that their independent and joint effects may be associated with small increases in BMI. PMID- 10480622 TI - Isolation, characterization, and chromosomal localization of the human ENSA gene that encodes alpha-endosulfine, a regulator of beta-cell K(ATP) channels. AB - Human alpha-endosulfine is an endogenous regulator of the beta-cell K(ATP) channels. The recombinant alpha-endosulfine inhibits sulfonylurea binding to beta cell membranes, reduces cloned K(ATP) channel currents, and stimulates insulin secretion from beta-cells. These properties led us to study the human ENSA gene that encodes alpha-endosulfine. Here, we describe the isolation, the partial characterization, and the chromosomal localization of the ENSA gene. The ENSA gene appears to be a 1.8-kb-long sequence that contains the transcription initiation site located 528 bp upstream of the initiation codon. The ENSA gene is intronless, and a single copy gene seems to be present in the genome. Finally, the ENSA gene co-localizes on human chromosome 14 (14q24.3-q31) with a locus for susceptibility to type 1 diabetes called IDDM11; thus, the ENSA gene represents an IDDM11 candidate. PMID- 10480623 TI - Human insulin receptor substrate-2: gene organization and promoter characterization. AB - Insulin receptor substrate-2 (IRS-2) belongs to a family of cytoplasmic adaptor proteins, which link insulin, IGF-1, and cytokine receptor tyrosine kinases to signaling pathways regulating metabolism, growth, and differentiation (1-3). IRS 2-deficient mice display all characteristics of type 2 diabetes, suggesting that dysfunction of the IRS-2 gene may contribute to the pathogenesis of human type 2 diabetes (4). Based on its progesterone inducibility, we have recently cloned and sequenced a full-length human IRS-2 cDNA containing an open reading frame (ORF) of 4,014 bp and 5'- and 3'-untranslated regions (UTRs) of 516 and 2,466 bp (5). Although the IRS-2 gene has previously been thought to lack introns within the coding region (6,7), the amino acid sequence predicted from our cDNA sequence differed at its very COOH-terminal end from an IRS-2 protein sequence derived from genomic IRS-2 sequences. Therefore, we carefully analyzed the genomic structure of the IRS-2 gene and found that the IRS-2 gene contains an intron that disrupts the ORF. Characterization of promoter and 5'-flanking regions of IRS-2 by sequencing, reporter gene assays, and chromatin structure analysis suggests that elements conferring progesterone inducibility are not located immediately upstream of the gene promoter. PMID- 10480624 TI - A polymorphism (K121Q) of the human glycoprotein PC-1 gene coding region is strongly associated with insulin resistance. AB - The genes responsible for insulin resistance are poorly defined. Plasma cell differentiation antigen (PC-1) glycoprotein inhibits insulin receptor signaling and is associated with insulin resistance. We describe here a novel polymorphism in exon 4 of the PC-1 gene (K121Q) and demonstrate that it is strongly associated with insulin resistance in 121 healthy nonobese (BMI <30 kg/m2) nondiabetic (by oral glucose tolerance test [OGTT]) Caucasians from Sicily. Compared with 80 KK subjects, Q allele carriers (n = 41, 39 KQ and 2 QQ) showed higher glucose and insulin levels during OGTT (P < 0.001 by two-way analysis of variance) and insulin resistance by euglycemic clamp (M value = 5.25 +/- 1.38 [n = 24] vs. 6.30 +/- 1.39 mg x kg(-1) x min(-1) [n = 49], P = 0.005). Q carriers had higher risk of being hyperinsulinemic and insulin resistant (odds ratio [CI]: 2.99 [1.28 7.0], P < 0.001). Insulin receptor autophosphorylation was reduced (P < 0.01) in cultured skin fibroblasts from KQ versus KK subjects. Skeletal muscle PC-1 content was not different in 11 KQ versus 32 KK subjects (33 +/- 16.1 vs. 17.5 +/ 15 ng/mg protein, P = 0.3). These results suggest a cause-effect relationship between the Q carrying genotype and the insulin resistance phenotype, and raise the possibility that PC-1 genotyping could identify individuals who are at risk of developing insulin resistance, a condition that predisposes to type 2 diabetes and coronary artery disease. PMID- 10480625 TI - Conservation of an insulin response unit between mouse and human glucose-6 phosphatase catalytic subunit gene promoters: transcription factor FKHR binds the insulin response sequence. AB - Because overexpression of the glucose-6-phosphatase catalytic subunit (G-6-Pase) in both type 1 and type 2 diabetes may contribute to the characteristic increased rate of hepatic glucose production, we have investigated whether the insulin response unit (IRU) identified in the mouse G-6-Pase promoter is conserved in the human promoter. A series of human G-6-Pase-chloramphenicol acetyltransferase (CAT) fusion genes was transiently transfected into human HepG2 hepatoma cells, and the effect of insulin on basal CAT expression was analyzed. The results suggest that the IRU identified in the mouse promoter is conserved in the human promoter, but that an upstream multimerized insulin response sequence (IRS) motif that is only found in the human promoter appears to be functionally inactive. The G-6-Pase IRU comprises two distinct promoter regions, designated A and B. Region B contains an IRS, whereas region A acts as an accessory element to enhance the effect of insulin, mediated through region B, on basal G-6-Pase gene transcription. We have previously shown that the accessory factor binding region A is hepatocyte nuclear factor-1, and we show here that the forkhead protein FKHR is a candidate for the insulin-responsive transcription factor binding region B. PMID- 10480627 TI - Elusive realities: a perspective on the Chicago retrovirus meeting. PMID- 10480628 TI - HIV type 1 tat gene heteroduplex mobility assay as a tool to establish epidemiologic relationships among HIV type 1-infected individuals. AB - Molecular biology techniques are increasingly used to study the molecular epidemiology of infectious diseases. Most of these methods are expensive and labor-intensive. The human immunodeficiency virus (HIV) has substantial genomic variation, such that HIVs from different individuals are genetically diverse, although mutation rates differ for distinct regions of the genome. Most studies of HIV linkage and molecular evolution have focused on env or gag regions. We show that heteroduplex mobility analysis of the first exon of the HIV tat gene provides a simple, rapid, inexpensive, and reliable discriminatory tool for the molecular differentiation of shared versus distinct HIV-1 quasispecies when epidemiologic relations need to be defined. tat, as a relatively conserved region, appears to be a better region than the more variable env region to establish HIV-1 epidemiological linkages. PMID- 10480626 TI - Genetic and physiologic analysis of the role of uncoupling protein 3 in human energy homeostasis. AB - By virtue of its potential effects on rates of energy expenditure, uncoupling protein 3 (UCP3) is an obesity candidate gene. We identified nine sequence variants in UCP3, including Val9Met, Val102Ile, Arg282Cys, and a splice site mutation in the intron between exons 6 and 7. The splice mutation results in an inability to synthesize mRNA for the long isoform (UCP3L) of UCP3. Linkage (sib pair), association, and transmission disequilibrium testing studies on 942 African-Americans did not suggest a significant effect of UCP3 on body composition in this group. In vastus lateralis skeletal muscle of individuals homozygous for the splice mutation, no UCP3L mRNA was detectable; the short isoform (UCP3S) was present in an increased amount. In this muscle, we detected no alterations of in vitro mitochondrial coupling activity, mitochondrial respiratory enzyme activity, or systemic oxygen consumption or respiratory quotient at rest or during exercise. These genetic and physiologic data suggest the following possibilities: UCP3S has uncoupling capabilities equivalent to UCP3L; other UCPs may compensate for a deficiency of bioactive UCP3L; UCP3L does not function primarily as a mitochondrial uncoupling protein. PMID- 10480629 TI - Genetic similarity of HIV type 1 subtype E in a recent outbreak among injecting drug users in northern Vietnam to strains in Guangxi Province of southern China. AB - To investigate the molecular epidemiology of a recent HIV-1 outbreak in northern Vietnam and its relation to the epidemic in surrounding areas, we analyzed 17 HIV positive blood specimens from 3 heterosexuals, 2 sexually transmitted disease patients, and 12 injecting drug users (IDUs), collected in 4 provinces near Hanoi in 1998. These were compared with the specimens from Ho Chi Minh City (n = 10) and An Giang Province (n = 10) in southern Vietnam and with published sequences from neighboring countries. Genetic subtyping based on the env C2/V3 sequences revealed that HIV-1 subtype E predominated throughout Vietnam in all risk populations; the exception was one typical United States-European-type HIV-1 subtype B detected in a patient in Ho Chi Minh City, the first case of HIV infection identified in Vietnam in 1990. The HIV-1 subtype E sequences identified in 9 of the 12 IDUs from northern provinces were closely related phylogenetically to those in IDUs in nearby Guangxi Province of China, and also shared a common amino acid signature downstream of the env V3 loop region. The low interperson nucleotide diversity among IDUs in northern Vietnam supports the view that HIV-1 subtype E was introduced recently among IDUs in northern Vietnam. These data indicate a linkage between HIV-1 circulating among IDUs in northern Vietnam and southern China, and suggest recent transborder introductions as the likely source of HIV-1 subtype E in northern Vietnam. PMID- 10480630 TI - Thalidomide stimulates T cell responses and interleukin 12 production in HIV infected patients. AB - We performed a placebo-controlled study to evaluate the effects of immunomodulatory treatment with thalidomide on HIV levels, TNF-alpha levels, and immune status of 31 HIV-infected individuals, after temporary suppression of viral replication with antiretroviral drugs. Treatment with a combination of zidovudine and lamivudine (ZDV/LMV) for 14 days resulted in a median decline in plasma viremia of 1.94 log10 RNA equivalents/ml. After discontinuation of ZDV/LMV, thalidomide therapy (200 mg/day for 4 weeks) did not retard the prompt return of HIV titers to the pretreatment levels, and had no effect on plasma levels of TNF-alpha. In contrast, thalidomide treatment resulted in significant immune stimulation. We observed increased levels of plasma soluble IL-2 receptor, soluble CD8 antigen, and IL-12 (p < 0.01 for all parameters), as well as increased cutaneous delayed-type hypersensitivity reactions to recall antigens (p < 0.01) in thalidomide-treated patients. These changes were associated with a median increase in HIV titer of 0.2 log10 RNA equivalents/ml in the thalidomide treated group (p < 0.05), which resolved after stopping the drug. Further studies were performed in vitro to elucidate the mechanism of thalidomide-induced immune stimulation. When purified T cells from HIV-infected individuals were stimulated by immobilized anti-CD3 in the presence of thalidomide, a costimulatory effect of the drug was observed, resulting in increased production of IL-2 and IFN-gamma, and increased T cell-proliferative responses. Further experiments showed that thalidomide increased IL-12 production by antigen-presenting cells in a T cell dependent manner. Our findings suggest a potential application for thalidomide as a novel immune adjuvant in HIV disease. PMID- 10480631 TI - The rabbit study: ritonavir and saquinavir in combination in saquinavir experienced and previously untreated patients. AB - Thirteen protease inhibitor-naive patients with HIV-1 infection, and 12 patients with a median of 58 months prior treatment with saquinavir (SQV) monotherapy, were treated with SQV (400 mg twice daily) and ritonavir (RIT, 500 mg twice daily) in a study designed to assess the effect of prior treatment with SQV monotherapy on the antiretroviral activity of RIT-SQV combination therapy. Median baseline viral load and CD4+ cell counts were 155,000 and 262,000 copies/ml and 333 and 225 cells/mm3 in the naive and experienced groups, respectively. Mean viral load changes at 24 weeks were -1.63 and -0.27 log copies/ml in the naive and SQV-experienced groups, respectively (intent-to-treat analysis). Baseline genotype by point mutation assay and sequencing in the SQV-experienced group was highly predictive of virological response. Eight of 11 SQV-experienced patients had evidence of phenotypic resistance to RIT at baseline, despite previous treatment with SQV only. There was strong correlation between phenotypic resistance to RIT and the presence of the L90M mutation. We conclude that prolonged prior treatment with saquinavir monotherapy may produce cross resistance to ritonavir and reduce the subsequent response to ritonavir saquinavir in combination. In this study, both phenotypic resistance to ritonavir and presence of the L90M mutation predicted the viral load response to ritonavir saquinavir. PMID- 10480632 TI - Rapid screening of phenotypic resistance to nevirapine by direct analysis of HIV type 1 reverse transcriptase activity in plasma. AB - Drug susceptibility testing for the clinical management of human immunodeficiency virus type 1 (HIV-1)-infected persons is often curtailed because such testing is expensive and time consuming. We describe a non-culture-based phenotypic assay for the rapid analysis of HIV-1 resistance to nevirapine. The assay measures the susceptibility of plasma reverse transcriptase (RT) activity to inhibition by nevirapine by using the PCR-based Amp-RT assay. Assay validation was made using two reference wild-type (WT) and six other nevirapine-resistant (>100-fold) HIV-1 isolates. Amp-RT IC50 values were found to correlate with those obtained by a conventional replication-based assay. The results also indicated that 50 microM nevirapine can be used in a single screening test to detect nevirapine resistance. Analysis of virus mixtures showed a detection threshold of 10% of nevirapine-resistant HIV-1 in a background of WT virus. To evaluate the assay on clinical samples, 30 plasma specimens collected longitudinally from 4 patients before and after treatment with nevirapine were analyzed, and results were compared with codon 181 genotypes. Preteatment samples and those obtained during the first 6 days of therapy (n = 21) were sensitive to nevirapine, and none had detectable Y181C mutation. Phenotypic resistance was seen in eight samples obtained after 1 week of treatment and was correlated with detection of the Y181C mutation. An increase in the level of phenotypic resistance was seen over time. These data validate this rapid and simple assay for monitoring phenotypic resistance to nevirapine. PMID- 10480633 TI - Partial resistance to infection by R5X4 primary HIV type 1 isolates in an exposed uninfected individual homozygous for CCR5 32-base pair deletion. AB - It is known that certain individuals remain persistently seronegative despite repeated exposure to HIV-1. Studies have shown that some exposed uninfected (EU) individuals who are homozygous for a 32-bp deletion in the CCR5 gene are resistant to infection with non-syncytium-inducing (R5) viruses. In the present investigation, we provide evidence that a highly exposed-uninfected individual with the CCR5 32-bp deletion (EUdelta32-1) also has partial resistance to syncytium-inducing (R5X4) HIV-1 viruses, when compared with unexposed-uninfected individuals with (UUdelta32-1 and UUdelta32-2) and without (UU-1 and UU-2) the 32 bp deletion. The partial resistance of EU cells was due neither to altered coreceptor expression, nor to specific mutation or deletion in the coding region of chemokine coreceptors CXCR4 and CCR3. While SDF-1, the ligand for CXCR4, blocked entry of R5X4 viruses to a similar extent in EUdelta32 and UUdelta32, there was a differential production of soluble factors by EUdelta32. Both CD4+ and CD8+ cells from EUdelta32-1 produced soluble factors that efficiently suppressed infection by HIV-1 R5X4 viruses when compared with supernatant from UUdelta32. These data provide evidence that additional soluble factors are involved in resistance to infection with R5X4 viruses. PMID- 10480634 TI - Extracellular HIV type 1 Tat protein induces CD69 expression through NF-kappaB activation: possible correlation with cell surface Tat-binding proteins. AB - The HIV-1 Tat protein, essential for HIV-1 gene expression and viral replication, is known to be secreted by infected cells and has pleiotropic effects on various cell functions. It seems that extracellular Tat may exert its functions on cellular targets by at least two different mechanisms, namely, by adsorptive endocytosis, and by a possible interaction with cell surface receptor(s). Here we report that extracellular Tat activates AIM/CD69 gene transcription through an NF kappaB-dependent pathway in the erythroleukemia cell line K562. Tat induces NF kappaB binding to DNA as a result of IkappaBalpha phosphorylation and degradation, which depend on the intracellular redox state. We found that the second Tat-coding exon is required for CD69 gene trans-activation, but not for HIV LTR gene transcription. Fluorescein-labeled Tat proteins were used to study cell surface binding sites and cellular uptake of the proteins. Full-length Tat protein has specific binding sites on the surface of K562 cells, whereas truncated Tat1-48, which is efficiently internalized by the cells, does not bind to the cell surface. Our results suggest that extracellular Tat may activate a cell surface-mediated pathway that induces intracellular signal transduction in K562 cells, leading to the activation of NF-kappaB and the transcription of NF kappaB-dependent genes, such as CD69. PMID- 10480635 TI - Human immunodeficiency virus type 1-specific cytotoxic T lymphocyte activity is inversely correlated with HIV type 1 viral load in HIV type 1-infected long-term survivors. AB - HIV-1-specific cytotoxic T cell (CTL) activity has been suggested to correlate with protection from progression to AIDS. We have examined the relationship between HIV-specific CTL activity and maintenance of peripheral blood CD4+ T lymphocyte counts and control of viral load in 17 long-term survivors (LTSs) of HIV-1 infection. Longitudinal analysis indicated that the LTS cohort demonstrated a decreased rate of CD4+ T cell loss (18 cells/mm3/year) compared with typical normal progressors (approximately 60 cells/mm3/year). The majority of the LTSs had detectable, variable, and in some individuals, quite high (>10(4) RNA copies/ml) plasma viral load during the study period. In a cross-sectional analysis, HIV-specific CTL activity to HIV Gag, Pol, and Env proteins was detectable in all 17 LTSs. Simultaneous analysis of HIV-1 Gag-Pol, and Env specific CTLs and virus load in protease inhibitor-naive individuals showed a significant inverse correlation between Pol-specific CTL activity and plasma HIV 1 RNA levels (p = 0.001). Furthermore, using a mixed linear effects model the combined effects of HIV-1 Pol- and Env-specific CTL activity on the viral load were significantly stronger than the effects of HIV-1 Pol-specific CTL activity alone on predicted virus load. These data suggest that the presence of HIV-1 specific CTL activity in HIV-1-infected long-term survivors is an important component in the effective control of HIV-1 replication. PMID- 10480637 TI - Federal support for training in laboratory animal medicine should be reinstated. PMID- 10480636 TI - Sequence of the env gene of some KwaZulu-Natal, South African strains of HTLV type I. AB - Phylogenetic analysis of HTLV-I suggests three main subtypes, namely, cosmopolitan, Central African, and Australo-Melanesian. HTLV-I is endemic in KwaZulu-Natal, South Africa. However, sequence data on the local strains are limited to the LTR region. The env gene of the local strain was amplified and sequenced from the peripheral blood of five seropositive individuals. Four had HTLV-I-associated myelopathy and one had infective eczema. The sequence analysis of the env gene showed a greater then 99% homology of the local strains. They were closely related to the North American strains (99.3%), followed by the Japanese strains (98.3-98.9%). Phylogenetic studies linked the local strains to the cosmopolitan subtype. This study provides new sequence data on the env gene of the local HTLV-I strain. PMID- 10480638 TI - Neuromuscular weakness in a baboon. PMID- 10480639 TI - Neurons and mechanisms of neuronal death in neurodegenerative diseases: a brief review. AB - BACKGROUND AND PURPOSE: Degenerative diseases of the central nervous system are a heterogenous group of slowly progressive disorders. A common feature of this group, which includes Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis, is gradual loss of specific populations of neurons. METHODS: A series of reports about neurodegenerative diseases and their relevant animal models, as well as a brief overview of the normal neuron and mechanisms of neuronal degeneration and death, is presented. CONCLUSION: Study of the aforementioned animal models, spontaneously occurring and experimentally induced, have provided important insights into the pathogenesis of these disorders and the development of effective therapeutic strategies. PMID- 10480640 TI - Experimental models of Parkinson's disease: insights from many models. AB - Toxin-induced and genetic experimental models have been invaluable in investigating idiopathic Parkinson's disease (PD). The neurotoxins--reserpine, 6 hydroxydopamine (6-OHDA), 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and methamphetamine--have been used to develop parkinsonian models in a wide variety of species. Both 6-OHDA and MPTP can replicate the neurochemical, morphologic, and behavioral changes seen in human disease. The unilateral 6-OHDA rat model is an excellent model for testing and determining modes of action of new pharmacologic compounds. The nonhuman primate MPTP-induced parkinsonian model has behavioral features that best approximate idiopathic PD. These induced and genetic models have been used to study the pathophysiology of the degenerating nigrostriatal system and to evaluate novel therapeutic strategies. Important differences within these models provide insights into various aspects of the dopaminergic phenotype and its role as a target in disease. These models provide an avenue to evaluate many anti-parkinsonian compounds, such as levodopa, which was first evaluated in an animal model and is the gold standard of parkinsonian treatment today. PMID- 10480641 TI - Pregnancy toxemia in the European ferret (Mustela putorius furo). AB - BACKGROUND AND OBJECTIVE: Pregnancy toxemia may lead to appreciable mortality among jills and their offspring. The objective of this report was to increase awareness of the disease, its likely cause, and practical prevention and treatment measures. METHODS: Ten cases of pregnancy toxemia were evaluated. Jills were in late gestation (mean, 38 days; range, 34 to 42 days) and had large litters (mean, 11.5 kits; range, 7 to 15 kits). RESULTS: The most common clinical signs of disease were lethargy, inappetence, dehydration, and excess shedding. Hematologic and clinical biochemical abnormalities included anemia (4 of 8 jills tested), hypoproteinemia (5 of 7), azotemia (7 of 7), hypocalcemia (5 of 6), hyperbilirubinemia (3 of 3), and high liver enzyme activities (6 of 6). Two jills were found dead; two jills were euthanized, six received supportive treatment, and cesarean section was performed on five. The three jills that survived tended to have less pronounced azotemia, hypoproteinemia, and liver enzyme activity increases and were not anemic. Hepatic lipidosis was observed grossly in all jills that died and was confirmed by histologic examination in four jills. CONCLUSIONS: Pregnancy toxemia in ferrets resembles metabolic diseases in several other animal species and requires aggressive treatment, including supportive care, nutritional supplementation, and cesarean section. Maintaining adequate nutrition and avoiding stress late in gestation may prevent the disease. PMID- 10480642 TI - Humoral immunity and protection of mice challenged with homotypic or heterotypic parvovirus. AB - BACKGROUND AND OBJECTIVES: Two serotypes of autonomously replicating parvoviruses infect laboratory mice. Genome regions coding for the nonstructural proteins of minute virus of mice [MVM] and mouse parvovirus [MPV] are almost identical, whereas capsid-coding sequences are divergent. We addressed these questions: Does humoral immunity confer protection from acute infection after challenge with homotypic or heterotypic parvovirus, and if it confers protection against acute MPV infection, does it also protect against persistent MPV infection? METHODS: Infant mice without maternal antibody or antibody to MVM or MPV and young adult mice given normal mouse serum or antibody to MVM or MPV were challenged with homotypic or heterotypic virus. In situ hybridization with target tissues was the indicator of infection. RESULTS: Humoral immunity failed to confer protection against acute heterotypic parvovirus infection. In passive transfer studies, MPV DNA was observed occasionally in lymph nodes, intestine, or the spleen of MPV challenged mice given homotypic antibody and kept for 6 or 28 days. Variable proportions of mice given MPV antibody and homotypic challenge had viral DNA in lymphoid tissues 56 days after virus inoculation. CONCLUSION: A mouse or colony that has sustained infection with MVM or MPV is probably fully susceptible to infection with the heterotypic virus. PMID- 10480643 TI - Antibody responses after Sendai virus infection and their role in upper and lower respiratory tract disease in rats. AB - BACKGROUND AND PURPOSE: Sendai virus infection in rats is an excellent model for studying development and role of host defenses throughout the respiratory tract after this infection. Therefore, development of serum antibody responses and disease were studied. METHODS: Forty-two anesthetized pathogen-free 3- to 4- week old LEW/NCr rats were inoculated intranasally with Sendai virus. At postinoculation days 0, 2, 3, 5, 8, 10, and 14, rats were euthanized by administration of a pentobarbital sodium overdose followed by exsanguination. Serum was obtained from all animals, and nasal wash and bronchoalveolar lavage specimens were collected during selected experiments. An ELISPOT assay was used to measure numbers of Sendai virus-specific antibody-forming cells in respiratory tract lymphoid tissue. RESULTS: Recovery from disease and clearance of virus from respiratory tract tissues coincided with development of serum antibody responses. Upper respiratory tract lymph nodes were the initial and major sites of appearance of antibody-forming cells. Immunoglobulin G was the predominant subtype of these cells during recovery from the infection and in rats resistant to infection. Passive transfer of antisera or specific IgG protected the lower but not the upper respiratory tract. CONCLUSIONS: Circulating components of immunity have a major role in resistance and recovery from disease in the lower respiratory tract, whereas local responses are likely involved in protection of the upper respiratory tract. Local lymphoid tissues are the major production sites of IgG, which contributes to resistance to and recovery from respiratory tract diseases. PMID- 10480644 TI - Lymphocyte subsets in neonatal and juvenile cats: comparison of blood and lymphoid tissues. AB - OBJECTIVE: To compare lymphocyte subpopulations in the blood and lymphoid tissues of normal kittens between 1 and 90 days of age. METHODS: Lymphocyte subsets within the blood, thymus, and lymph node of 24 normal kittens were quantified by use of two-color fluorescence flow cytometry and were compared at 1, 23, 46, or 90 days after birth. RESULTS: Blood B and T lymphocytes increased over the 90-day postnatal period. The CD4+ and CD8+ sub-populations of T lymphocytes increased. However, CD8+ lymphocytes increased more than did CD4+ lymphocytes, resulting in reduced CD4-to-CD8 ratio. By 23 days of age, similar but more abrupt changes in the CD4-to-CD8 ratio occurred in the thymus and lymph nodes, coinciding with the highest thymus-to-body weight ratio and gradual increase in mature thymocytes expressing a pan-T lymphocyte marker. CONCLUSIONS: Postnatal thymopoiesis in the domestic cat favors production of mature CD8+ T lymphocytes over CD4+ T lymphocytes. This coincides with the emergence of CD8+ lymphocytes in the lymph node and precedes a more gradual increase in CD8+ cells in the blood. Therefore, the ontogeny of these effectors of cell-mediated immunity could be interrupted by infective agents that target lymphoid tissues of the neonate. PMID- 10480645 TI - Effect of access to a running wheel on behavior of C57BL/6J mice. AB - BACKGROUND AND PURPOSE: To evaluate how and when mice run on a running wheel and how ad libitum access to the wheel affect behavior, feed intake, and weight gain. METHODS: Seventeen 2-month-old C57BL/6J mice had access to the wheel, whereas 19 control mice did not. After 3 to 6.5 weeks, behavior was video-recorded over 24 h for each mouse. RESULTS: Experimental mice ran an average 2 km/24 h in 114 min. Highest running activity took place at the onset of darkness. Experimental mice spent 22 min more feeding on the cage floor than did control mice. These times were deducted from those for all other behaviors: 74 min from resting time, 39 min from climbing and feeding on the cage lid, 14 min from locomotion on the cage floor, and 10 min from grooming. In relative figures, deduction from sleeping time was only 9%, whereas climbing time was halved. CONCLUSIONS: Climbing on the cage lid has a similar circadian rhythm as does wheel running and high-energy expenditure. Because experimental mice climbed less, their weight gain and feed intake were similar to those of control mice. Thus, wheel running can substitute for other forms of energy-consuming behaviors and vice versa. PMID- 10480646 TI - Complications of ileal cannulation in cats. AB - BACKGROUND AND PURPOSE: The anatomic position of the ileum is such that use of simple, noninvasive sampling techniques is difficult. Sampling of ileal contents by use of a cannula provides information about those contents-digestibility and fermentation patterns; however, complications with use of cannulas in horses and dogs have been reported. Therefore, cats were chosen as a laboratory model in which to study ileal cannulation. METHODS: Simple T-piece cannulas were surgically placed in the ileum of six adult cats. RESULTS: Postsurgical complications included cannula displacement and leakage, with subsequent abscessation and skin inflammation. Owing to these complications, the cannulas were removed from four of the six cats. Similar complications developed after removal of the cannulas. In the only attempt to collect a sample, obtaining a 0.5 ml sample of ileal fluid from one cannula took 3 h. Of the six cats, four died or were euthanized. CONCLUSION: Use of cannulas is not a viable technique of collecting ileal fluid samples in cats. PMID- 10480647 TI - Effects of citrated whole blood transfusion in response to hemorrhage. AB - BACKGROUND AND PURPOSE: Standard treatment for massive hemorrhage in dogs is infusion of whole blood or of packed red blood cells with fresh frozen plasma if whole blood is not available. Although most whole blood is collected using a citrate-based anticoagulant, knowledge of citrate's relevant non-anticoagulant effects is not widespread. Citrate's anticoagulant activity is achieved through chelation of divalent metal cations (e.g., magnesium, calcium), which may exacerbate cardiovascular and metabolic insults attributable to hemorrhage. METHODS: Blood pressures, gas tensions, metabolites, and electrolytes; myocardial metabolites, pressures, and contractility; cardiac output; and left cranial descending and circumflex coronary artery flows were measured in 21 anesthetized dogs after hemorrhage was induced by collection of blood into a citrated reservoir to mean arterial pressure of 45 mm Hg for approximately 60 min (until arterial lactate concentration was 7.0 mmol/L), followed by a 1-h transfusion and 2 h of maintenance. RESULTS: Arterial ionized calcium concentration, total peripheral resistance, and myocardial function decreased significantly during hemorrhage. All aforementioned responses but myocardial function continued to decrease during the initial 20 min of transfusion, then began to recover. Total peripheral resistance and end-systolic elastance were the only factors significantly related to calcium concentration. CONCLUSION: Transfusion with citrated whole blood may significantly alter calcium concentration, negatively affecting myocardial and vascular function. PMID- 10480648 TI - Evaluation of the micronucleus test in peripheral blood erythrocytes by use of the splenectomized model. PMID- 10480649 TI - Experimentally induced infection of gerbils with cilia-associated respiratory bacillus. PMID- 10480650 TI - Alpha-mannosidosis in a guinea pig. PMID- 10480651 TI - Evaluation of FVB/N mice as recipients for transgenic embryos. PMID- 10480652 TI - Technique for continuous monitoring of salivary cortisol concentration in pigs. PMID- 10480653 TI - Experimental hyperlipidemia prevents the protective effect of ischemic preconditioning on the contractility and responsiveness to phenylephrine of rat isolated stunned papillary muscle. AB - This study was designed to establish a hyperlipidemic diet (significant increase in the cholesterol and triglycerides blood levels, but without atherogenic changes in heart muscle and coronary vessels) and to investigate the influence of experimental hyperlipidemia on the effects of ischemic preconditioning (PC) of rat-isolated papillary muscle on the time course of contractility during simulated ischemia and reperfusion and responsiveness to phenylephrine under such a condition. The animals were divided in four experimental groups: standard diet fed control group (SD), SD underwent ischemic preconditioning (SD-PC), hyperlipidemic diet-fed group (HLD) and HLD underOFFt PC (HLD-PC). Force of contraction (Fc), velocity of contraction (+dF/dt), and velocity of relaxation ( dF/dt) were measured. HLD preparations were more sensitive to ischemia then SD ones. PC, performed by 5-min perfusion with no-substrate solution gassing with 95% N2/5% CO2 in the presence of fast electrical stimulation, and 10 min of reperfusion with normal solution and rate of stimulation, significantly increased the resistance of isolated cardiac tissues to simulated ischemia in SD-PC group, but not in HLD-PC group. Negative inotropic action of phenylephrine occured in SD group of preparations after simulated-ischemia/reperfusion period was also prevented by PC. Therefore, we conclude that experimental hyperlipidemia significantly influenced the function of rat heart muscle including the higher sensitivity to ischemia and different reaction to the same PC procedure. PMID- 10480654 TI - The potentiating effect of genistein on the relaxation induced by isoproterenol in rat aortic rings. AB - In rat aortic rings, the mechanism of potentiating effect of genistein, a tyrosine kinase inhibitor, on the relaxation induced by isoproterenol was examined. Pretreatment of the aortic rings by genistein, but not by daidzein, an inactive analogue of genistein, potentiated the relaxation induced by isoproterenol. Genistein also potentiated the relaxation induced by forskolin, an activator of guanylyl cyclase, and dibutyryl cyclic AMP. In addition, theophylline, an inhibitor of phosphodiesterase, potentiated the relaxation induced by isoproterenol and forskolin. Theophylline partly inhibited the potentiation of isoproterenol-induced relaxation by genistein while it completely inhibited the potentiation of forskolin-induced relaxation by genistein. Iberiotoxin, an inhibitor of Ca-activated K (KCa) channels, partly inhibited the isoproterenol-induced relaxation and the potentiating effect of genistein on the relaxation induced by isoproterenol. Quinacrine (an inhibitor of phospholipase A2), alpha-naphthoflavone (an inhibitor of cytochrome P-450 enzymes), and 8 methoxypsoralen (an inhibitor of cytochrome P-450 enzymes), partly inhibited the potentiating effect of genistein on the isoproterenol-induced relaxation, but metyrapone (an inhibitor of cytochrome P-450 enzymes), indomethacin (an inhibitor of cyclooxygenase), and AA861 (an inhibitor of 5-lipoxygenase) did not. These results suggest that the potentiation of isoproterenol-induced relaxation by genistein may be related to the activities of phosphodiesterase, KCa channels, and cytochrome P-450 enzymes. PMID- 10480655 TI - Biphasic positive inotropic actions of doxorubicin in isolated guinea pig hearts: relation to Ca2+ release from the sarcoplasmic reticulum. AB - Doxorubicin (30 microM) caused a biphasic (early and late phase) positive inotropic effect. A high concentration (200 microM) of this agent caused a stronger early phase, but a weaker late phase. The early phase caused by the high concentration of doxorubicin was significantly reduced by cyclopiazonnic acid or thapsigargin, although it was not altered by ryanodine, verapamil, or nifedipine. The late phase in the presence of the high concentration of doxorubicin was slightly enhanced by cyclopiazonic acid, verapamil or nifedipine, and markedly enhanced by ryanodine. At the end of the 4-hr experimental period in the presence of the high concentration of doxorubicin, the positive inotropic effects of CaCl2 were completely diminished. Verapamil almost completely restored the action of the doxorubicin. PMID- 10480656 TI - Pharmacological characterization of metamizol-induced relaxation in phenylephrine precontracted rabbit thoracic aorta smooth muscle. AB - Metamizol produced a dose- and time-dependent relaxation in rabbit thoracic aorta smooth muscle that was precontracted by phenylephrine. Such a relaxation was not observed with indomethacin, which is also a nonsteroidal anti-inflammatory drug. The relaxing effect of metamizol was independent of the presence of vascular endothelium. Tetraethylammonium (a calcium-activated potassium channel inhibitor), glybenclamide (an ATP-dependent potassium channel inhibitor), indomethacin (a cyclooxygenase inhibitor), and methylene blue (a soluble guanylate cyclase inhibitor) did not have any effect on metamizol-induced relaxation response. Metamizol did not produce any relaxation effect on aortic smooth muscle when KCl (30, 60, and 117 mM KCl) was used instead of phenylephrine to precontract the preparation. Ouabain (a Na-K ATPase pump inhibitor) showed a dose-dependent inhibition on metamizol's relaxation response. However, in potassium-free medium, which is an alternative way to block the Na-K ATPase pump, no inhibition in metamizol-induced relaxation response was observed. When metamizol was incubated for 2 h in organ-bath conditions before evaluating its relaxing effect, it produced a relatively faster relaxation, indicating that the relaxing effect of metamizol is produced by one of its (active) spontaneous degradation products (possibly 4-methylaminoantipyrine). PMID- 10480657 TI - Effect of bradykinin and its antagonist on survival time after coronary artery occlusion in rats. AB - The present study was conducted to examine the effect of bradykinin and bradykinin 2 receptor antagonist on survival time in rats with coronary artery ligation for 15 min and continuously. We also evaluated the heart rate and blood pressure responses in the presence and absence of bradykinin and its antagonist. Bradykinin treatment (4 microg and 8 microg/kg IV) significantly (p < 0.05) increased the survival time of rats compared with saline-treated rats with coronary artery ligation for 15 min and continuously. The heart rate and blood pressure responses were significantly (p < 0.001) altered in the presence of coronary artery ligation. Bradykinin antagonist treatment (4 microg/kg IV) abolished the effect of bradykinin and thus reduced the survival time of rats with coronary artery ligation. The mean value of survival time between saline treated and bradykinin antagonist- plus bradykinin-treated rats did not differ significantly (p > 0.05). PMID- 10480658 TI - Effects of single intraperitoneal injections of an extract of Ginkgo biloba (EGb 761) and its terpene trilactone constituents on barbital-induced narcosis in the mouse. AB - A mouse model of barbital-induced narcosis was used to examine the effects of single intraperitoneal injections of an extract of Ginkgo biloba (EGb 761), an extract devoid of terpene trilactones (CP 205), and three terpene trilactone constituents of the extract (ginkgolides A and B, bilobalide). Administration of sodium barbital (180 mg/kg, IP) to the mice caused narcosis, measured as a loss in righting reflex. Single injections of EGb 761 (25 and 50 mg/kg), given 60 min prior to sodium barbital, significantly shortened barbital-induced sleeping time, whereas these same doses of CP 205 were ineffective. Single injections of ginkgolide B (1 mg/kg) and bilobalide (2 and 5 mg/kg) significantly shortened sleeping time, whereas ginkgolide A was ineffective. The effects of ginkgolide B and bilobalide were reflected as increases in latency to onset of sleep and those of EGb 761, ginkgolide B, and bilobalide were correlated with decreases in the number of mice that slept. At the behavioral level, these potent in vivo effects of EGb 761, ginkgolide B, and bilobalide resemble those of certain antidepressants. At the molecular level, it is hypothesized that interactions with the picrotoxinin/TBPT site of GABA-regulated Cl- channels of the CNS may be involved. This information appears useful in explaining the clinically observed "vigilance-enhancing" and "antidepressant-like" actions of EGb 761. PMID- 10480659 TI - Nitrated nonivamide displaying a drawback of proton's role in capsaicin associated sensory and neuronal activities. AB - Nitrated nonivamide (NVANO)-induced triad hypotension, and biphasic bradycardia at 0.25-1.0 mg/kg (IV) was inhibited by capsazepine (1.0 mg/kg, IV), atropine (1.0 mg/kg, IV), and vagotomy in rats. NVANO also elicited a hypotensive spinal reflex at 5.0 mg/kg (IA). In the isolated rat vagus, NVANO (10.0-100.0 microM) revealed a sensory C-spike inhibition and membrane depolarization. NVANO (5.0 microM)-induced calcium influx in the isolated rat dorsal root ganglion cells (DRGs) was diminished by capsazepine (10.0 microM). In the isolated guinea pig atria, NVANO (1.0-50.0 microM)-induced positive inotropic and chronotropic activities were antagonized by capsazepine (1.0-10.0 microM) and human calcitonin gene-related peptide(8-37) (hCGRP(837); 0.1-1.0 microM). PMID- 10480660 TI - Effects of cimetidine and its metal complexes on nitroblue tetrazolium and ferricytochrome c reduction by superoxide radicals. AB - The effects of cimetidine, a potent histamine-H2-receptor antagonist and a good OH scavenger, on the kinetics of ferricytochrome c and NBT reduction by superoxide anions were studied. The drug dose-dependently inhibited ferricytochrome c and NBT reduction by O2- radicals, generated either in xanthine oxidase system or photochemically or directly by KO2. The inhibitory effect of cimetidine remained unchanged in the presence of catalase or mannitol. Cimetidine and its complexes with Cu(II) and Fe(III) ions inhibited ferricytochrome c and NBT reduction even when metal chelators were added to the reaction medium. The results suggest the reaction of cimetidine with O2-radicals. PMID- 10480661 TI - Renal effects of 2-mercaptoacetyl-L-leucyl-L-phenylalanine, a novel selective inhibitor of neutral endopeptidase 24.11 (Neprilysin): comparison with SQ 28,603. AB - The effects of 2-mercaptoacetyl-L-leucyl-L-phenylalanine (MA-LF) on the activity of neutral endopeptidase and on renal hemodynamics and excretory function were investigated in experiments in vitro and in vivo. In vitro studies showed that the compound effectively inhibited purified bovine kidney neutral endopeptidase (Ki = 0.012 microM), while having slight influence on the activity of angiotensin I converting enzyme (Ki = 0.14 microM). In experiments on normal anesthetized rats (thiobutabarbital sodium salt, 100 mg/kg), IV administration of MA-LF (20 and 60 mg/kg) produced a dose-dependent increase in absolute rate and fractional excretion of sodium (+324% and +299%, respectively) and urinary flow rate (+261%), but did not change renal and systemic hemodynamics. Renal excretory effects of the new compound were comparable to those of the selective neutral endopeptidase inhibitor SQ 28,603. These results demonstrate that MA-LF is a potent neutral endopeptidase inhibitor with prominent natriuretic and diuretic properties. PMID- 10480663 TI - The treatment of tardive dyskinesia--a systematic review and meta-analysis. AB - This systematic review aimed to collate randomized controlled trials (RCTs) of various interventions used to treat tardive dyskinesia (TD) and, where appropriate, to combine the data for meta-analysis. Clinical trials were identified by electronic searches, handsearches and contact with principal investigators. Data were extracted independently by two reviewers, for outcomes related to improvement, deterioration, side-effects and drop out rates. Data were pooled using the Mantel-Haenzel Odds Ratio (fixed effect model). For treatments that had significant effects, the number needed to treat (NNT) was calculated. From 296 controlled clinical trials, data were extracted from 47 trials. For most interventions, we could identify no RCT-derived evidence of efficacy. A meta analysis showed that baclofen, deanol and diazepam were no more effective than a placebo. Single RCTs demonstrated a lack of evidence of any effect for bromocriptine, ceruletide, clonidine, estrogen, gamma linolenic acid, hydergine, lecithin, lithium, progabide, seligiline and tetrahydroisoxazolopyridinol. The meta-analysis found that five interventions were effective: L-dopa, oxypertine, sodium valproate, tiapride and vitamin E; neuroleptic reduction was marginally significant. Data from single RCTs revealed that insulin, alpha methyl dopa and reserpine were more effective than a placebo. There was a significantly increased risk of adverse events associated with baclofen, deanol, L-dopa, oxypertine and reserpine. Meta-analysis of the impact of placebo (n=485) showed that 37.3% of participants showed an improvement. Interpretation of this systematic review requires caution as the individual trials identified tended to have small sample sizes. For many compounds, data from only one trial were available, and where meta-analyses were possible, these were based on a small number of trials. Despite these concerns, the review facilitated the interpretation of the large and diverse range of treatments used for TD. Clinical recommendations for the treatment of TD are made, based on the availability of RCT-derived evidence, the strength of that evidence and the presence of adverse effects. PMID- 10480662 TI - Improvement of cisplatin-induced emesis and delayed gastric emptying by KB-R6933, a novel 5-HT3 receptor antagonist. AB - The effects of a novel 5-hydroxytryptamine3 (5-HT3) receptor antagonist, KB R6933, 6-amino-5-chloro-1-isopropyl-2-(4-methyl-1-piperazinyl)-benzimidazole dimaleate, on emesis and delayed gastric emptying induced by cisplatin were assessed in experimental models. Prophylactic intravenous or oral treatment with KB-R6933 prolonged the latent period until the first emetic episode and decreased the number of emetic episodes induced by cisplatin in ferrets. KB-R6933 immediately inhibited the subsequent emesis when administered to the ferrets which exhibited established vomiting after administration of cisplatin. In rats treated with cisplatin, the gastric emptying rate was significantly reduced. KB R6933 reversed the reduction of gastric emptying induced by cisplatin. These results suggest that KB-R6933 is an antiemetic agent, and could improve the cisplatin-induced delay of gastric emptying. PMID- 10480664 TI - Antipsychotic drug effects on motor activation measured by functional magnetic resonance imaging in schizophrenic patients. AB - Brain function and laterality in schizophrenia were investigated by means of a simple motor task with a self-generated left-hand sequential finger opposition (SFO) using a whole-brain high-speed (100 ms per slice) functional imaging technique. Neuroleptic-naive, acutely ill schizophrenic patients were compared to schizophrenic patients under stable neuroleptic medication and matched controls. The goal was to evaluate both the motor function in first-episode patients and possible effects of different neuroleptic treatments on functional MRI results. Forty patients satisfying ICD 10 criteria (F20.x) for schizophrenia and sex- and age-matched healthy volunteers participated in this study. All subjects underwent fMRI examinations on a conventional 1.5 T MR unit. The primary sensorimotor cortex and the high-order supplementary motor area (SMA) were evaluated. There was a close similarity in the activation of the primary and high-order (SMA) sensorimotor areas between first-episode schizophrenic patients and controls. In contrast, a significant reduction in the overall blood oxygen level dependent (BOLD) response was seen in sensorimotor cortices (contra- and ipsilateral) in schizophrenic patients under stable medication with typical neuroleptics. This effect was not present in patients treated with atypical antipsychotics. Both antipsychotic treatments, however, led to a significant reduction in activation of the SMA region compared to controls and neuroleptic-naive subjects. Thus, the present study provides no evidence for the localized involvement of the primary motor cortex or the SMA as a relatively stable vulnerability marker in schizophrenia. There is, however, strong evidence that neuroleptics themselves influence fMRI activation patterns and that there are major differences between typical neuroleptics and atypical antipsychotics. PMID- 10480665 TI - Symptom dimensions in old-age schizophrenics. Relationship to neuropsychological and motor abnormalities. AB - In most factor analytical studies of schizophrenic symptomatology, a three-factor solution was found. The aim of this study was to investigate symptomatological dimensions in old age and to clarify whether the dimensions correlate differently with neuropsychological and motor parameters. One hundred and thirty-one DSM-III R chronic schizophrenics (mean age 68 years) were assessed using SANS, SAPS, a neuropsychological test battery and motor scales. Exploratory and confirmatory factor analyses yielded a model with three dimensions (negative, disorganized, paranoid), two of which (negative, disorganized) showed different correlations with neuropsychological and motor phenomena. Thus, three symptomatological dimensions could also be demonstrated in a chronic, old-age schizophrenic sample. The pathophysiological significance of the different correlations with neuropsychological and motor parameters should be clarified in neuroimaging and neuropathological studies. PMID- 10480666 TI - Latent structures underlying schizophrenic symptoms: a five-dimensional model. AB - Latent structures of schizophrenic phenomenology were examined over the course of the illness in 100 newly-admitted patients. We compared the results of a confirmatory factor analysis (CFA) on ten competing models that had between zero and five dimensions using data assessed by the Positive and Negative Syndrome Scale (PANSS) at both the acute and chronic phases of the disease. The present findings did not support the two-dimensional construct of positive and negative symptoms in either the acute or the chronic phase of the illness. In the acute phase, a three- (positive, negative, and relational dimensions), four- (positive, negative, disorganization, and relational dimensions), and five-dimensional model (positive, negative, disorganization, excitement, and relational dimensions) fit the data relatively well. In contrast, in the chronic stable phase, only the five dimensional model adequately fits the data. The present findings suggest that further investigation of the validity of the five-dimensional model over the course of the illness is necessary. PMID- 10480667 TI - Increased stroop facilitation effects in schizophrenia are not due to increased automatic spreading activation. AB - Studies using the single trial Stroop task consistently reveal increased reaction time (RT) facilitation effects among schizophrenia patients. One possible mechanism underlying this effect is increased automatic spreading activation in semantic networks. The current study was designed to test this hypothesis. We administered the Stroop task and two semantic priming tasks to the same subjects. Patients showed greater Stroop RT facilitation than controls, no evidence of increased semantic priming at short stimulus onset asynchronies (SOAs), and reduced semantic priming at long SOAs. In addition, abnormal Stroop performance was related to the severity of Disorganization symptoms. These results are inconsistent with the spreading activation hypothesis. Alternative hypotheses regarding the source of Stroop task performance deficits in schizophrenia are discussed. PMID- 10480668 TI - Psychopathology and cognition in schizophrenia spectrum disorders: the role of depressive symptoms. AB - The cognitive correlates of five symptom dimensions based on PANSS ratings were examined in a group of 50 recent onset psychotic patients, using both objective and subjective cognitive measures. We were particularly interested in the depression dimension, since it has not been studied extensively thus far. The depression dimension showed a high number of correlations with both objective and subjective cognitive measures, such as problems with simple and divided attention, psychomotor slowing and subjectively experienced distractibility, overload and diminished attentional control. The other dimensions, including negative symptoms, have less cognitive correlates. It is possible that previous studies based on a three-dimensional model confounded correlates of negative symptoms with correlates of depressive symptoms. The results of this study suggest the need for more research into the mechanisms underlying the relationship between depressive symptoms and cognitive functioning in schizophrenia, and that patients with depressive symptoms are less efficient in information processing, but can compensate by investing more mental effort. Because subjective cognitive measures were related to mental effort in previous research, they can be a useful tool in future research. PMID- 10480669 TI - California Verbal Learning Test practice effects in a schizophrenia sample. AB - Practice effects on the California Verbal Learning Test (CVLT; >Delis et al., 1987), a measure of new learning and memory, were evaluated in a sample of patients with schizophrenia who were administered the CVLT at baseline, week 10, and week 14 in the context of a study of the effects of a non-pharmacological intervention on psychiatric status. Large effects attributable to prior exposure to the test were evident at weeks 10 and 14. These effects indicate that caution must be exercised in interpreting serial performances on this commonly used test, whether in research or clinical circumstances. Additionally, although the exact nature of the learning involved is unclear, the influence of prior exposure on later performance reveals considerable retention over time of new information in this sample of persons with schizophrenia. PMID- 10480670 TI - Accuracy of the seven subtest WAIS-R short form in chronic schizophrenia. AB - The accuracy of the WAIS-R seven subtest short form (Ward, L.C., 1990. Prediction of Verbal, Performance and Full Scale IQs from seven subtests of the WAIS-R. J. Clin. Psychol. 46, 436-440) was examined for predicting IQs of 73 inpatients diagnosed with schizophrenia. Results indicated that 93% of the estimated Full Scale IQs were within +/-5 points of their actual scores. Using Wechsler's (1981) seven category intelligence classification, the level of agreement on the Full Scale IQ was 84% for the standard WAIS-R and the seven subtest short form. This abbreviated Wechsler Scale may be used with schizophrenic patients when only general estimates of intellectual functioning are required. PMID- 10480671 TI - The prolactin response to fenfluramine in schizophrenia is associated with negative symptoms. AB - We examined the serotonergic system in drug-free inpatients [schizophrenia (n=28) and schizoaffective (n=7)] by administering a challenge dose of oral dl fenfluramine (fenfluramine) in a controlled angiocatheter study. Seventeen of these patients were also randomized to a placebo condition in addition to receiving fenfluramine. Response to fenfluramine as reflected by changes in serum prolactin and cortisol were examined by repeated measures ANOVA, as well as Area Under the Curves (AUCs). The prolactin response, but not the cortisol response, was significantly correlated with measures of negative symptoms. PMID- 10480672 TI - A neural network model of general olfactory coding in the insect antennal lobe. AB - A central problem in olfaction is understanding how the quality of olfactory stimuli is encoded in the insect antennal lobe (or in the analogously structured vertebrate olfactory bulb) for perceptual processing in the mushroom bodies of the insect protocerebrum (or in the vertebrate olfactory cortex). In the study reported here, a relatively simple neural network model, inspired by our current knowledge of the insect antennal lobes, is used to investigate how each of several features and elements of the network, such as synapse strengths, feedback circuits and the steepness of neural activation functions, influences the formation of an olfactory code in neurons that project from the antennal lobes to the mushroom bodies (or from mitral cells to olfactory cortex). An optimal code in these projection neurons (PNs) should minimize potential errors by the mushroom bodies in misidentifying the quality of an odor across a range of concentrations while maximizing the ability of the mushroom bodies to resolve odors of different quality. Simulation studies demonstrate that the network is able to produce codes independent or virtually independent of concentration over a given range. The extent of this range is moderately dependent on a parameter that characterizes how long it takes for the voltage in an activated neuron to decay back to its resting potential, strongly dependent on the strength of excitatory feedback by the PNs onto antennal lobe intrinsic neurons (INs), and overwhelmingly dependent on the slope of the activation function that transforms the voltage of depolarized neurons into the rate at which spikes are produced. Although the code in the PNs is degraded by large variations in the concentration of odor stimuli, good performance levels are maintained when the complexity of stimuli, as measured by the number of component odorants, is doubled. When excitatory feedback from the PNs to the INs is strong, the activity in the PNs undergoes transitions from initial states to stimulus-specific equilibrium states that are maintained once the stimulus is removed. When this PN-IN feedback is weak the PNs are more likely to relax back to a stimulus-independent equilibrium state, in which case the code is not maintained beyond the application of the stimulus. Thus, for the architecture simulated here, strong feedback from the PNs onto the INs, together with step-like neuronal activation functions, could well be important in producing easily discriminable odor quality codes that are invariant over several orders of magnitude in stimulus concentration. PMID- 10480673 TI - Quantitative trait loci associated with short-term intake of sucrose, saccharin and quinine solutions in laboratory mice. AB - The goal of this study was simultaneously to map two genetic loci which, collectively, have a large effect on intake of sucrose, saccharin and quinine solutions in mice. These loci had been previously identified using long-term measurements with the traditional two-bottle test, but the present study used a short-term, one-bottle test. Intake of distilled water, 100 mM sucrose, 10 mM sodium saccharin and 1.1 mM quinine HCl over 6 h was measured on two occasions from a non-deprived group of 61 male and 72 female F2 mice derived from a cross of the C57BL/6J and DBA/2J mouse strains and used to detect quantitative trait loci (QTL). DNA from each animal was typed for polymorphisms in anonymous microsatellite markers on mouse chromosomes 4 and 6. Saccharin and sucrose relevant QTL were detected on distal chromosome 4 and a quinine relevant QTL was detected on medial/distal chromosome 6 in the region of Prp. The location of these QTL and the proportion of phenotypic variance they accounted for were similar to those arrived at following previous determinations using the two bottle test. Measurement stability for the three gustatory phenotypes was high, product-moment correlation coefficients between first and second determinations varying between approximately 0.80 for sucrose and saccharin and 0.73 for quinine. QTL parameters assessed independently for first and second presentations of sucrose and saccharin were stable, but the location of the quinine QTL differed between presentations. The present experiment illustrates the utility of a 6 h fluid intake test in the mapping of Sac and Qui loci. The short duration of the test provides a simple means of measuring variation in gustatory processes and the discovery that these loci influence short-term as well as long-term fluid intake extends understanding of the mechanism of gene action. PMID- 10480674 TI - Induction of salivary gurmarin-binding proteins in rats fed gymnema-containing diets. AB - Gymnema sylvestre, a tropical plant, contains gurmarin that selectively suppresses sucrose responses of the chorda tympani nerve in rats and mice. We investigated preference for taste solutions and saliva composition in rats fed a diet containing this plant (gymnema diet). Preference for 0.01 M sucrose and a mixture of 0.03 M sucrose and 0.03 mM quinine-HCl significantly decreased at 1-2 days after the start of the gymnema diet and subsequently returned closely to the control levels within about a week. There was no significant change in preference for NaCl, monosodium glutamate and quinine-HCl during feeding trials. Submandibular saliva of rats fed the gymnema diet for 4 and 14 days showed an inhibitory effect on immunoreaction between gurmarin and antigurmarin serum. Analyses using electrophoresis and affinity chromatography indicated that the saliva contains gurmarin binding proteins with molecular weights of 15, 16, 45, 60 and 66 kDa. These results suggest that reduction of preference for sucrose was probably caused by gurmarin contained in the gymnema diet and subsequent restoration of the preference may be due to suppression of the effect of gurmarin by salivary gurmarin-binding proteins induced by the gymnema diet. PMID- 10480675 TI - Taste confusions following gymnemic acid rinse. AB - The effect of a gymnemic acid (GA) rinse, which simulated a sweet-taste deficit, was measured on human taste perception and identification. Taste ratings showed that GA reduced the intensities of sucrose and aspartame to 14% of pre-rinse levels; over the recovery interval of 30 min, these values increased linearly to 63% of the pre-rinse levels. Repeated presentations of a set of 10 stimuli (five primarily or partly sweet--sucrose, aspartame, and NaCl-sucrose, acid-sucrose and quinine-sucrose mixtures; and five nonsweet--NaCl, KCl, Na glutamate (MSG), quinine HCl and citric acid) for identification following water and GA rinses produced 'taste confusion matrices' (TCMs). Correct identification of the sweet tasting stimuli was reduced by 23% in presentations closely following the GA rinse, an effect that dissipated with time. Most misidentifications involved sucrose and mixtures containing sucrose. In a second TCM experiment, GA was presented frequently within each session to maintain the sweet taste deficit, which revealed itself as specific confusions. Rinsing with GA impaired discriminability of sweet-nonsweet pairs of stimuli but enhanced discriminability of the aspartame-(NaCl-sucrose) pair. GA had no effect on discriminability of nonsweet stimulus pairs. The results suggest that specific error patterns in the TCM could be used to identify quality-specific taste disorders. PMID- 10480676 TI - Oral irritant properties of piperine and nicotine: psychophysical evidence for asymmetrical desensitization effects. AB - Using a bipolar rating scale, human subjects rated the intensity of irritation sensation evoked by repeated application of piperine (75 p.p.m.) or nicotine (0.12%) to one side of the dorsal surface of the tongue. The intensity of irritation elicited by repeated application of piperine significantly increased, while irritation elicited by repeated nicotine significantly decreased. We additionally tested if nicotine or piperine desensitized the tongue. After either piperine or nicotine was repeatedly applied to one side of the tongue, a 5 or 10 min rest period ensued, followed by re-application of piperine or nicotine to both sides of the tongue. Subjects were asked to choose which side of the tongue gave rise to a stronger irritation in a two-alternative forced choice (2-AFC) paradigm. In addition, they gave separate ratings of the intensity of irritation on the two sides of the tongue. When piperine was applied bilaterally after unilateral pretreatment with piperine and a 10 min rest period, subjects consistently chose the non-pretreated side to yield stronger irritation and assigned significantly higher ratings to that side, indicative of piperine self desensitization. A similar self-desensitization effect was found when bilateral application of nicotine followed unilateral treatment with nicotine and a 5 min rest period. Unilateral treatment with piperine also reduced nicotine-evoked irritation on the pretreated side (cross-desensitization), but treatment with nicotine did not affect piperine-evoked irritation. This asymmetrical cross desensitization pattern is similar to that observed between capsaicin and nicotine and constitutes an additional similarity between piperine and capsaicin. PMID- 10480677 TI - Alteration of perceived fragrance of essential oils in relation to type of work: a simple screening test for efficacy of aroma. AB - The perceptional change of fragrance of essential oils is described in relation to type of work, i.e. mental work, physical work and hearing environmental (natural) sounds. The essential oils examined in this study were ylang ylang, orange, geranium, cypress, bergamot, spearmint and juniper. In evaluating change in perception of a given aroma, a sensory test was employed in which the perception of fragrance was assessed by 13 contrasting pairs of adjectives. Scores were recorded after inhaling a fragrance before and after each type of work, and the statistical significance of the change of score for 13 impression descriptors was examined by Student's t-test for each type of work. It was confirmed that inhalation of essential oil caused a different subjective perception of fragrance depending on the type of work. For example, inhalation of cypress after physical work produced a much more favorable impression than before work, in contrast to orange, which produced an unfavorable impression after physical work when compared with that before work. For mental work, inhalation of juniper seemed to create a favorable impression after work, whereas geranium and orange both produced an unfavorable impression then. From these studies, together with those conducted previously with lavender, rosemary, linalool, peppermint, marjoram, cardamom, sandalwood, basil and lime, we thus concluded that the sensory test described here might serve not only as a screening test for efficacy of aroma but also as a categorized table for aroma samples which can act as a reference to each other. PMID- 10480678 TI - Actographic analysis of the effects of an esterase inhibitor on male moth responses to sex pheromone. AB - The effects of 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP), a trifluoromethyl ketone that inhibits antennal esterases, on male Mamestra brassicae responses to the main pheromone component have been investigated using an actograph. This actograph used a movement detector based on the Doppler effect. The signal from the detector was digitalized and analysed on a PC microcomputer to quantify male activity. When added to the air flowing through the observation chamber, OTFP inhibited the responses of male moths to the pheromone. The number of males responding to the pheromone and the intensity of the response were decreased by OTFP. The latency of the response was increased and its duration decreased. These effects on the kinetics of the behavioural response cannot be directly correlated to the inhibition of pheromone catabolism by OTFP and other targets must be involved. The high level of inhibition of behaviour observed in presence of OTFP demonstrates the interest of trifluoromethyl ketones as mating disruption agents for pest control. PMID- 10480679 TI - Olfactory discrimination ability and odor structure-activity relationships in honeybees. AB - Using the training procedure introduced by von Frisch in 1919, we tested the ability of free-flying honeybees to discriminate a conditioning odor from an array of 44 simultaneously presented substances. The stimuli included homologous series of aliphatic alcohols, aldehydes and ketones, isomeric forms of some of these substances, as well as several terpenes and odor mixtures, and thus comprised stimuli of varying degrees of structural similarity to any conditioning odor. We found (i) that the honeybees significantly distinguished between 97.0% of the 1848 odor pairs tested, thus showing an excellent discrimination performance when tested in a free-flying situation with an array of structurally related substances; (ii) a significant negative correlation between discrimination performance and structural similarity of odorants in terms of differences in carbon chain length with all aliphatic substance classes tested; (iii) that both the position and type of a functional group also affected discriminability of odorants in a substance class-specific manner; and (iv) striking similarities in odor structure-activity relationships between honeybees and human and nonhuman primates tested previously on a subset of substances employed here. Our findings demonstrate that the similarities found in the structural organization of the olfactory systems of insects and vertebrates are paralleled by striking similarities in relative discrimination abilities. This strongly suggests that similar mechanisms of odor coding and discrimination may underlie olfaction in vertebrates and insects. PMID- 10480680 TI - Selective inhibition of sweetness by the sodium salt of +/-2-(4 methoxyphenoxy)propanoic acid. AB - The purpose of this study was to determine the degree to which the sodium salt of +/-2-(4-methoxyphenoxy)propanoic acid (Na-PMP) reduced sweet intensity ratings of 15 sweeteners in mixtures. Na-PMP has been approved for use in confectionary/frostings, soft candy and snack products in the USA at concentrations up to 150 p.p.m. A trained panel evaluated the effect of Na-PMP on the intensity of the following 15 sweeteners: three sugars (fructose, glucose, sucrose), three terpenoid glycosides (monoammonium glycyrrhizinate, rebaudioside A, stevioside), two dipeptide derivatives (alitame, aspartame), two N sulfonylamides (acesulfame-K, sodium saccharin), two polyhydric alcohols (mannitol, sorbitol), 1 dihydrochalcone (neohesperidin dihydrochalcone), one protein (thaumatin) and one sulfamate (sodium cyclamate). Sweeteners were tested at concentrations isosweet with 2.5, 5, 7.5 and 10% sucrose in mixtures with two levels of Na-PMP: 250 and 500 p.p.m. In addition, the 15 sweeteners were tested either immediately or 30 s after a pre-rinse with 500 p.p.m. Na-PMP. In mixtures, Na-PMP at both the 250 and 500 p.p.m. levels significantly blocked sweetness intensity for 12 of the 15 sweeteners. However, when Na-PMP was mixed with three of the 15 sweeteners (monoammonium glycyrrhizinate, neohesperidin dihydrochalcone and thaumatin), there was little reduction in sweetness intensity. Pre-rinsing with Na-PMP both inhibited and enhanced sweetness with the greatest enhancements found for monoammonium glycyrrhizinate, neohesperidin dihydrochalcone and thaumatin, which were not suppressed by Na-PMP in mixtures. The mixture data suggest that Na-PMP is a selective competitive inhibitor of sweet taste. The finding that pre-treatment can produce enhancement may be due to sensitization of sweetener receptors by Na-PMP. PMID- 10480681 TI - The taste of monosodium glutamate (MSG), L-aspartic acid, and N-methyl-D aspartate (NMDA) in rats: are NMDA receptors involved in MSG taste? AB - Monosodium glutamate (MSG) is believed to elicit a unique taste perception known as umami. We have used conditioned taste aversion assays in rats to compare taste responses elicited by the glutamate receptor agonists MSG, L-aspartic acid (L Asp), and N-methyl-D-aspartate (NMDA), and to determine if these compounds share a common taste quality. This information could shed new light upon the receptor mechanisms of glutamate taste transduction. Taste aversions to either MSG, L-Asp or NMDA were produced by injecting rats with LiCl after they had ingested one of these stimuli. Subsequently, rats were tested to determine whether they would ingest any of the above compounds. The results clearly show that a conditioned aversion to MSG generalized to L-Asp in a dose-dependent manner. Conversely, rats conditioned to avoid L-Asp also avoided MSG. Conditioned aversions to MSG or L Asp generalized to sucrose when amiloride was included in all solutions. Importantly, aversions to MSG or L-Asp did not generalize to NMDA, NaCl or KCl, and aversions to NMDA did not generalize to MSG, L-Asp, sucrose or KCl. These data indicate that rats perceive MSG and L-Asp as similar tastes, whereas NMDA, NaCl and KCl elicit other tastes. The results do not support a dominant role for the NMDA subtype of glutamate receptors in taste transduction for MSG (i.e. umami) in rats. PMID- 10480682 TI - Olfactory event-related potentials in young and elderly adults: evaluation of tracking task versus eyes open/closed recording. AB - The purpose of the present study was to evaluate olfactory event-related potentials (OERPs) elicited by amyl acetate from subjects performing a visuomotor tracking task compared with the no-task conditions of eyes open and eyes closed. Task condition did not produce any reliable effects for any amplitude measure. Task type weakly influenced only P2 latency. Elder adults evinced smaller P2 and N1/P2 amplitudes and longer N1 and P2 latencies than young adults. The results suggest that tracking task performance is not necessary to obtain robust OERPs from normal subjects of a wide age range. PMID- 10480683 TI - The influence of early experience with vanillin on food preference later in life. AB - A study with 133 adults, who had been breast-fed or bottle-fed after birth, shows that neonatal experience with vanilla influences preferences for other foods in later life. PMID- 10480684 TI - Interactions of familial and hormonal risk factors for large bowel cancer in women. AB - BACKGROUND: Family history of colorectal cancer has been consistently associated with an increased personal risk of this disease. Since evidence suggests that hormones are related to colon cancer risk in women, the effect of family history on large bowel incidence may be modified according to endogenous and exogenous hormone levels. METHODS: We analysed data from a population-based case-control study of female colorectal cancer to evaluate family history and cancer risk. Cases (n = 702) were female residents of Wisconsin with a new diagnosis of colorectal cancer, identified through a statewide tumour registry. Controls (n = 2274) were randomly selected from lists of licensed drivers and from rosters of Medicare beneficiaries. All relative risks (RR) were adjusted for age, body mass index, smoking and alcohol history, education, and use of hormone replacement therapy. RESULTS: Compared with women who reported no history of cancer in a first degree relative, women with a family history had an RR of 2.07 (95% confidence interval [CI]: 1.60-2.68). Regardless of which parent was affected, risks were increased about twofold, while sibling history was associated with about a 50% increase in risk. Risk was greater if more than one family member was affected (RR 3.65, 95% CI: 1.81-7.37). The association between family history and risk was stronger for colon cancer than for rectal cancer. There were no indications that exogenous hormonal factors, notably hormone replacement use, modified these risks. There was a suggestion that high parity attenuated the risks associated with family history (P = 0.07). CONCLUSIONS: These results confirm that family history of colorectal cancer is associated with a doubling of risk for large bowel cancer in women; some histories were associated with greater risk. This relation was not substantially different among subgroups of women with varying exogenous and endogenous hormone exposures. PMID- 10480685 TI - Ulcerative colitis and colorectal cancer: a follow-up study in Fukuoka, Japan. AB - BACKGROUND: Our goal was to study the higher death rate and the causes of such deaths among ulcerative colitis (UC) patients in the Japanese population, and to compare our findings in such cases with those for Crohn's disease (CD). METHODS: In all, 174 UC (male/female: 54/120) and 66 CD (34/32) patients who were registered for the research promotion programme in Fukuoka prefecture (1971-1981) were traced up to the end of 1994. The standardized mortality ratios (SMR) were calculated based on the death rates of the Japanese population by age, sex and calendar year. RESULTS: The overall follow-up rate was 96.7%. Among the UC patients, the SMR for all causes were 0.84 (95% CI: 0.11-4.31) for men; 1.05 (95% CI: 0.08-4.69) for women; and 0.94 (95% CI :0.09-4.50) for both sexes combined. When excluding deaths due to colorectal cancer, the SMR for the same groups were 0.43, 0.94 and 0.67, respectively. The SMR for both sexes were 1.82 (95% CI: 0.17 5.96) for malignant neoplasms and 9.93 (95% CI: 4.67-17.3) for colorectal cancer. Patients who died from colorectal cancer showed onset at a younger age (mean: 25.5 years) as well as a longer disease course of UC (mean: 17.0 years). Regarding the CD patients, the SMR for all causes were 1.75 (95% CI: 0.15-5.75) for both sexes. Most deaths were caused by gastrointestinal complications. CONCLUSIONS: An excess mortality from colorectal cancers was indicated in the UC patients, especially in males. The overall SMR in male UC patients decreased by 50% when the deaths from colorectal cancer were excluded. The excess mortality in those with CD over UC patients was attributed to gastrointestinal complications rather than malignant diseases. Some carcinogenic factors therefore seem most likely to exist in the pathogenesis of UC. PMID- 10480686 TI - Cigarette smoking and male lung cancer risk with special regard to type of tobacco. AB - BACKGROUND: The mortality rate from lung cancer (LC) increased sharply in Spain between 1957 and 1986. This increase has been related to a previous increase in cigarette smoking. Certain features of cigarette smoking which were frequent among Spanish smokers (use of black tobacco and use of cigarettes without filter) have been related to a higher risk of LC. METHODS: A hospital-based case-control study was conducted between December 1986 and June 1990. The 325 male patients with lung cancer included in the study (cases) were compared with 325 age-matched male controls without LC. Occupation and lifetime tobacco consumption were requested using a structured questionnaire. The LC odds ratios (OR) and 95% CI were estimated with multiple logistic regression. RESULTS: Lung cancer risk increased with cigarette consumption and duration of the habit. After adjusting for lifetime cigarette consumption and for socioeconomic level, LC risk was greater among black tobacco smokers than among exclusive blond tobacco smokers (OR = 5.0, 95% CI: 2.0-12.7); LC risk among long-term (> or =20 years) filter tipped cigarette users was lower compared to all other smokers (OR = 0.4, 95% CI: 0.2-0.7). CONCLUSIONS: The main results of the study (a higher LC risk among black tobacco users than in exclusive blond tobacco users, and a lower LC risk among long-term filter-tipped cigarette smokers than all other smokers) have been consistent with previous case-control studies and with ecologic studies which took into account past exposure levels. PMID- 10480687 TI - Physical activity and risk of lung cancer. AB - BACKGROUND: Physical activity has been proposed to decrease lung cancer risk; however, few data are available. Further, no studies have examined specific kinds and intensities of activities. METHODS: We conducted a prospective cohort study among 13 905 male Harvard University alumni (mean age, 58.3 years), free of cancer. Men reported their walking, stair climbing and participation in sports or recreation on baseline questionnaires in 1977, and the occurrence of lung cancer on follow-up questionnaires in 1988 and 1993. Death certificates were obtained for decedents through 1992 to determine lung cancers not previously reported. RESULTS: During follow-up, 245 men developed lung cancer. Adjusting for age, cigarette smoking, and body mass index, the relative risks of lung cancer associated with <4200, 4200-8399, 8400-12 599 and > or =12 600 kJ/week of estimated energy expenditure at baseline were 1.00 (referent), 0.87 (95% CI: 0.64 1.18), 0.76 (95% CI: 0.52-1.11), and 0.61 (95% CI: 0.41-0.89), respectively; P trend = 0.0008. Similar trends were observed among non-smokers or former smokers in 1977 (82.7% of men) as well as among those who smoked >20 cigarettes a day in 1977 (8.0%), although the findings in the latter group were not statistically significant, possibly due to the small number. Walking, climbing stairs and participating in activities of at least moderate intensity (> or =4.5 MET, or multiples of resting metabolic rate) were each inversely associated with lung cancer risk, independent of the other activity components. However, light intensity activities (<4.5 MET) did not predict lung cancer risk. CONCLUSIONS: These data indicate that physical activity may be associated with lower risk of lung cancer among men. An energy expenditure of 12 600 kJ/week, achievable by perhaps 6-8 hours of at least moderate intensity physical activity, may significantly lower risk. Further studies are required to confirm these observations. PMID- 10480688 TI - Case-control study of thyroid cancer in Northern Italy: attributable risk. AB - BACKGROUND: The percentage of thyroid cancer cases attributable to specific risk factors can be calculated to focus preventive strategies. The per cent population attributable risks (PAR) for thyroid cancer were estimated in relation to history of benign thyroid diseases, history of radiotherapy, residence in endemic goitre areas and selected indicators of a poor diet, using data from a case-control study conducted between 1986 and 1992 in Northern Italy. METHODS: Cases were 399 histologically confirmed incident thyroid cancers and controls were 617 patients, admitted to hospital for a wide range of acute, non-neoplastic, non-hormone related diseases. The PAR were computed on the basis of multivariate odds ratios (OR) and on the distribution of risk exposure among cases, assuming they are representative of the general population of cases. RESULTS: A history of benign thyroid disease accounted for 18.9% of cases, radiotherapy for 1.2%, residence for > or =20 years in endemic goitre areas for 2.4% of cases, and their combination for 21.7% of thyroid cancer cases; selected indicators of a poor diet accounted for 40.9% of thyroid cancer cases in this population. The combination of all factors considered explained over 57% of thyroid cancer cases in both sexes. The estimates for thyroid-related conditions were higher in women than men, whereas the opposite was true for dietary indicators. The overall PAR were somewhat higher in people aged > or =45 years (63.8%) than in younger subjects, and for follicular (69.1%) rather than papillary (53.7%) cancers. CONCLUSIONS: Exposure to a few simply identified and potentially modifiable risk factors or indicators (benign thyroid disease, residence in endemic goitre area and a poor diet) explained about 60% of thyroid cancer cases in this Italian population, indicating the theoretical scope for prevention. PMID- 10480689 TI - Association of childhood cancer with factors related to pregnancy and birth. AB - BACKGROUND: It has been hypothesized that risk factors of childhood cancers may already operate during the prenatal and neonatal period. Results of previous epidemiological studies have been inconsistent. METHODS: During 1992-1997 a large case-control study on childhood cancers and a variety of potential risk factors was conducted in Germany. Cases were ascertained by the German Childhood Cancer Registry. Each case was matched to a population-based control of the same age and gender, sampled from the district where the case lived at the date of diagnosis. For the analyses, 2358 cases and 2588 controls were available. RESULTS: Risk of childhood acute leukaemia increased with maternal age < or =20 years at time of delivery (odds ratio [OR] = 1.9, 95% CI: 1.1-3.2), lower (<2500 g: OR = 1.7, 95% CI: 1.1-2.8) and higher birthweight (>4000 g: OR = 1.4, 95% CI: 1.0-1.8, P < 0.05), and hormonal treatment because of infertility (OR = 1.6, 95% CI: 1.0-2.5, P < 0.05). No associations were seen for parental smoking habits, maternal alcohol consumption during pregnancy and fetal losses. Parity was associated only with subgroups of acute leukaemias. Regarding non-Hodgkin's lymphoma we observed an elevated OR for lower birthweight and heavy maternal smoking during pregnancy (>20 cigarettes/day) and a decreased OR for children with one or two siblings. Only a few significant findings were seen for the different groups of solid tumours. CONCLUSIONS: Overall, only weak associations were identified and the evaluated risk factors operating during the neonatal and prenatal period account at most for only a small proportion of childhood cancers. PMID- 10480690 TI - Increasing inequality in ischaemic heart disease morbidity among employed men in Denmark 1981-1993: the need for a new preventive policy. AB - BACKGROUND: In the mid 1980s European governments committed themselves to the WHO goal 'reduced inequality in health by year 2000' according to which inequality in health should be reduced by 25% by the year 2000. The study aim is to estimate the time trend in relative risk due to ischaemic heart disease (IHD) morbidity in employment status groups in Denmark in the period from 1981 to 1993 and to recommend a strategy to reduce inequality in health. MATERIAL AND METHODS: The study dealt with change in relative risk of IHD in main employment status AND groups as measured in three successive cohorts. The cohorts were defined as all METHODS: gainfully employed men in Denmark as of 1 January 1981, 1986 and 1991, respectively. Information on employment was retrieved for the three previous years. The cohorts were followed for first admissions with IHD as the principal cause during 5, 5, and 3 years respectively. RESULTS: Managers and white collar workers had an average or low and decreasing relative risk while male blue collar workers had a high and increasing relative risk. Thus the social inequality in IHD is rapidly increasing. Some occupational groups are known to be at high risk. Some of these high-risk groups, such as bus drivers, even have an increasing relative risk. CONCLUSIONS: The general health education has been successful in the prevention of IHD in the high-status groups but has failed to reduce the risk among blue collar workers. Preventive measures against IHD should focus on occupational groups at high, increasing risk and the measures should tailor to their 'subculture.' PMID- 10480691 TI - Serum magnesium and ischaemic heart disease: findings from a national sample of US adults. AB - BACKGROUND: Animal and human data suggest that magnesium may play an important role in ischaemic heart disease. Few prospective epidemiological studies have related serum magnesium concentrations to mortality from ischaemic heart disease (IHD) or all-causes. METHODS: Data from the National Health and Nutrition Examination Survey Epidemiologic Followup Study were used to examine the association between serum magnesium concentration, measured between 1971-1975, and mortality from IHD or all-causes in a national sample of 25-74-year-old participants followed for about 19 years. RESULTS: The analytical samples for IHD and all-cause-mortality included 12 340 and 12 952 participants, respectively (1005 IHD deaths, 2637 IHD deaths or hospitalizations, 4282 total deaths). Hazard ratios for IHD mortality from proportional hazards analysis comparing the second (1.59-<1.68 mEq/l), third (1.68-<1.77 mEq/l), and fourth (> or =1.77 mEq/1) quartiles of serum magnesium concentration with the lowest quartile were 0.79 (95% CI: 0.58-1.08), 0.66 (95% CI: 0.47-0.93), 0.69 (95% CI: 0.52-0.90), respectively. For all-cause mortality, hazards ratios were 0.82 (95% CI: 0.72 0.93), 0.84 (95% CI: 0.73-0.96), 0.85 (95% CI: 0.75-0.95). No significant interactions between serum magnesium concentration and age, sex, race, and education were observed. CONCLUSION: Serum magnesium concentrations were inversely associated with mortality from IHD and all-cause mortality. PMID- 10480692 TI - Psychological stress and incidence of ischaemic heart disease. AB - BACKGROUND: We assessed the relationship between psychological stress and ischaemic heart disease (IHD) incidence in a population of 868 men over a 10-year follow-up period. METHODS: In 1981, 869 men aged 42-60, free from IHD and living around Quebec City completed a questionnaire assessing the presence of psychological stress in different areas of their life. They also underwent a medical examination and provided information on IHD risk factors. From 1981 to 1991, the incidence of IHD events was ascertained. The relationship between 13 stress dimensions and IHD incidence was investigated using Cox regression while controlling for important IHD risk factors. Cross-sectional analyses were also performed to investigate the relationship between stress dimensions and IHD risk factors. RESULTS: Between 1981 and 1991, 79 men (9%) experienced a first IHD event. The following risk factors were associated with the risk of IHD: age, (rate ratio (RR) = 1.93, 95% CI: 1.21-3.09), hypertension (RR = 1.90, 95% CI: 1.22-2.98), triglycerides (RR = 1.87, 95% CI: 1.19-2.95) and high density lipoprotein (HDL) cholesterol (RR = 1.64, 95% CI: 1.05-2.55). After controlling for risk factors, not one of the psychological stress dimensions significantly altered the risk of IHD. CONCLUSIONS: While confirming the influence of hypertension, age, triglycerides and HDL cholesterol on IHD incidence, this study suggests that there is no important connection between the explored stress dimensions and IHD incidence. It is unlikely that this lack of association is due to the stress questionnaire since the 13 stress dimensions were rigorously developed through independent evaluation of the questions by three specialists and many statistically significant relationships were observed between stress dimensions and IHD risk factors. PMID- 10480693 TI - Benefits of leisure-time physical activity on the cardiovascular risk profile at older age. AB - BACKGROUND: Intensity, frequency and duration of physical activity may contribute in different ways to the maintenance of cardiovascular health. Their relative importance may also change at different stages in life and this should be taken into account for activity recommendations. METHODS: The relationship of frequency and duration of leisure-time physical activities with cardiovascular risk factors was studied in 4942 male and 5885 female participants aged 50-69, of the German Cardiovascular Prevention Study (1984-1991). RESULTS: After adjustment for several possible confounders, women with modest levels (2-12 times per month, 0.5 2 h per week) of moderate-to-vigorous activity (> or =5 kcal/kg/h) had significantly lower systolic blood pressure (-1.8%), resting heart rate (-3.1%) and body mass index (-3.2%) values than sedentary women. Beneficial differences increased with frequency and duration of activity. Light activities (3-4.5 kcal/kg/h), conducted > or =5 times a week, were significantly associated with favourable lower diastolic blood pressure (-1.4%), resting heart rate (-2.3%) among women, and body mass index (women -2.9%, men -2.2%) among both genders. Recommended activity levels (> or =5 times, > or =3.5 h weekly) were associated with a lower prevalence of multiple risk factors (odds ratio [OR] = 0.55, 95% CI: 0.41-0.75 for men and OR = 0.44 95% CI: 0.31-0.63 for women). CONCLUSIONS: For sedentary elderly, even less physical activity than currently recommended, is likely to improve the cardiovascular risk profile. PMID- 10480694 TI - Associations of education with cardiovascular risk factors in young adults: the Cardiovascular Risk in Young Finns Study. AB - BACKGROUND: Low educational level is associated with an increased risk of coronary heart disease. The aim of the present study was to examine the relationships between education and common cardiovascular risk factors in young adults. METHODS: Trends in conventional risk factors of young adults aged 21, 24, 27 and 30 years in 1992 (n = 443) were examined across the educational groups as part of a 12-year follow-up study, the Cardiovascular Risk in Young Finns Study. Education was determined as participants' own educational level and as parental years of schooling. RESULTS: In males, subject's own education was related inversely and independently of parental school years to serum total and low density lipoprotein (LDL) cholesterol concentration, smoking and body mass index. In females, participant's own educational level associated inversely with smoking and physical inactivity. Parental school years was associated inversely and independently of one's own educational level with serum total and LDL cholesterol values and waist-hip ratio in females. In both genders, parental education was a stronger determinant of diet (butter use) than one's own educational level. CONCLUSIONS: The least educated young adults have adopted a more adverse lifestyle than the more educated. The risk factor profile in young adulthood, especially in females, is still affected by parental education. The influences of one's own and parental educational level on vascular risk profile should be taken into consideration when planning public health campaigns among young adults. PMID- 10480695 TI - Increased blood pressure in adolescents who were small for gestational age at birth: a cohort study in Brazil. AB - BACKGROUND: This paper studies the relationship between birthweight for gestational age and blood pressure in adolescents aged 14-15 years in southern Brazil. METHODS: A sample of 1076 adolescents belonging to a cohort of over 6000 children born in 1982 in Pelotas, southern Brazil, was studied in 1997. All households in a sample of 25% of the city's census tracts were visited and all adolescents born in 1982 were interviewed, weighed, and their blood pressures were measured twice. Data from the adolescents were linked to the database through their names and dates of births. RESULTS: High diastolic and systolic pressure (defined as >95th percentile) were significantly more frequent among adolescents who were born below the 10th percentile of birthweight for gestational age. No association was found between high blood pressure and low birthweight or preterm births. In a multiple linear regression analysis, the association between birthweight for gestational age and blood pressure was not statistically significant after adjusting for age, sex, skin colour and family income. However, when the current body mass index and height were added to the model both diastolic and systolic pressure were significantly associated with birthweight for gestational age, and adolescents who were small for gestational age at birth presented a mean elevation of 3.08 mmHg for diastolic pressure and 2.89 mmHg for systolic pressure. CONCLUSIONS: There is an inverse association between birthweight for gestational age and blood pressure during adolescence. This association, however, is only disclosed when the negative confounding effect of the body mass index is controlled for. The same association is not found when the effects of birthweight and gestational age on blood pressure are analysed separately. It appears therefore that the elevation of blood pressure during adolescence only occurs when there was intra-uterine growth retardation. PMID- 10480696 TI - Selecting subjects for ultrasonographic screening for aneurysms of the abdominal aorta: four different strategies. AB - BACKGROUND: We studied whether the effectiveness of ultrasound screening for abdominal aortic aneurysms could be increased by preselecting high-risk subjects, based on the presence of risk indicators for the disease. METHODS: In a population-based screening programme for abdominal aortic aneurysms among 5328 subjects living in Rotterdam, The Netherlands, we studied four different strategies to select subjects for ultrasound screening of the abdominal aorta, based on risk indicators for abdominal aortic aneurysm disease. Risk indicators used in each strategy were entered in a logistic regression model to predict the probability of an individual having an abdominal aortic aneurysm. Using several cutoff values for the probability of a subject having an aneurysm for each strategy, we estimated the proportion of subjects that should be referred for ultrasound screening and the proportion of aneurysms that would be diagnosed by each strategy (sensitivity). RESULTS: When a probability of 1.5% of having an aneurysm is chosen as the cutoff point above which ultrasound screening is indicated, the proportion of subjects that would be referred for screening ranged from 36% (first strategy) to approximately 50% (other strategies), while 80% (first strategy) to approximately 94% (other three strategies) of all aneurysms would be detected. CONCLUSION: Effectiveness in screening for abdominal aortic aneurysms can be increased by selecting subjects by means of a short medical questionnaire, filled out by the screening candidates, including questions on medical history. PMID- 10480697 TI - Risk factors for the increasing caesarean section rate in Southeast Brazil: a comparison of two birth cohorts, 1978-1979 and 1994. AB - BACKGROUND: Brazil has the highest caesarean section (CS) rate in the world (36.4% in 1996). METHODS: Risk factors for increasing CS rate were studied in two population-based cohorts of singleton live births in families residing in the municipality of Ribeirao Preto, State of Sao Paulo, Southeast Brazil. The first comprised births from June 1978 to May 1979 (6750 births-one-year survey) and the second births from May to August 1994 (2846 births-4-month survey). Multiple unconditional logistic regression modelling was used to control for confounding. RESULTS: The CS rate rose from 30.3% in 1978-1979 to 50.8% in 1994. In 1978-1979, socioeconomic, reproductive and demographic variables, and health service factors were associated with CS rate. In 1994, only reproductive, demographic and health service factors remained associated, e.g. hour of delivery (from 7 a.m. to 12 p.m.), attendance by the same physician for prenatal care and delivery, > or =4 prenatal visits, maternal age > or =30 years, 1-3 previous live births and birthweight 3500-3999 g. CONCLUSION: Caesarean section in Brazil is widely performed for non-medical reasons in which physician convenience plays an important role. There is an urgent need for public health interventions to reduce the CS rate in Brazil, mainly directed towards cultural beliefs and physician behaviour. PMID- 10480698 TI - Low birthweight and prematurity in relation to paternal factors: a study of recurrence. AB - BACKGROUND: The importance of paternal determinants in the occurrence of low birthweight and prematurity is not well known. We investigated these outcomes in siblings and paternal half siblings as a function of changes in putative external determinants between two births in fathers who had experienced the birth of a premature and/or low birthweight (PTB/LBW) infant. METHODS: All fathers who, between 1980 and 1992, had an infant born before 37 completed weeks' gestation or weighing <2500 g and a following child were studied. We identified 14 147 pairs of siblings from Danish national registers. The recurrence risk was studied in three sub-cohorts defined by the outcome in the index child (PTB only, PTB/LBW, LBW only). We estimated the recurrence risk in the younger sibling according to changes of female partner, municipality type, occupation, and father's social status. RESULTS: The overall recurrence risk was 16.7% for preterm delivery and 16.8% for LBW. Changing female partner was, as expected, associated with a reduction in the recurrence risk for both outcomes (RR = 0.40; 95% CI: 0.27-0.60 for preterm delivery and RR = 0.38; 95% CI : 0.26-0.56 for LBW). None of the other studied factors was associated with changes in the recurrence risk. Fathers who changed partner had offspring with similar birthweight and gestational length between the three sub-cohorts, while a difference was evident in offspring to fathers whose female partner was unchanged. CONCLUSIONS: We did not identify any paternal factor of importance in the occurrence of LBW and preterm delivery. PMID- 10480699 TI - A modified method for the epidemiological analysis of registry data on infants with multiple malformations. AB - BACKGROUND: Infants with multiple malformations are important in birth defect epidemiology and malformation monitoring because human teratogens have often caused complex malformations. Various methods for the analysis of multimalformed infants have been tried. METHOD: By using data from four large registries of congenital malformations, 5256 infants were identified with two or more among 73 selected malformations. Pairwise associations between malformations were detected by multiple logistic regression analyses, and putative confounders (programme, maternal age, autopsy, etc.) were controlled for. For each significant pairwise association, further analyses were performed in order to find associations with a possible third malformation. RESULTS: The importance of controlling for several confounders was demonstrated. Several well-known associations were found, which supports the technique used. The interpretation of three-way associations was discussed. Results from the present study were compared with those obtained using some other methods. CONCLUSIONS: Different confounders can cause biased associations. The method presented in the paper takes this into consideration and is therefore more likely than previously used techniques to give unbiased information on the clustering of different malformations among multimalformed infants. PMID- 10480700 TI - Trisomy 18 in Kuwait. AB - BACKGROUND: Trisomy 18 (Edwards' syndrome, T18) is the second most common trisomy in man. We describe 118 children with regular T18 who were ascertained clinically and cytogenetically in the Kuwait Medical Genetics Centre during 1980-1997. METHODS: Ascertainment of T18 cases was performed shortly after birth. Chromosomal studies were carried out in addition to other relevant investigations. To investigate the factors associated with T18, a case-control study was carried out with 131 normal healthy newborns. Studied factors included maternal and paternal age, birth order, abortion, associated malformation, and survival. Multiple logistic regression analysis was used to adjust for confounding between variables. RESULTS: There was a preponderance of females among T18 cases (female:male ratio 2.1:1). The majority of T18 cases (53%) died before the second week of life. The most common associated anomalies were: congenital heart (38.1%) and gastrointestinal (25.4%). Multiplicity of malformations was also observed. Significant seasonal variation in T18 cases was detected with a peak in spring. Of the 118 T18 cases, 59 were delivered during 1994-1997 (average overall T18 birth prevalence rate 8.95 per 10 000 live births [95% CI: 6.66-11.23]). Concerning maternal age, 30.5% of the T18 cases' mothers were > or =35 years compared to 10.7% in the control group. The difference was statistically significant, P = 0.002. Logistic regression analysis showed that maternal age >30 years was a significant risk factor for T18, after adjusting for confounding with paternal age. Paternal age and abortion were not found to be significant risk factors. CONCLUSION: Trisomy 18 birth prevalence rate is high in Kuwait with advanced maternal age as a significant risk factor. PMID- 10480701 TI - Prevalence of asthma, aspirin intolerance, nasal polyposis and chronic obstructive pulmonary disease in a population-based study. AB - BACKGROUND: Remarkable overlap exists in symptoms between asthma and chronic obstructive pulmonary disease (COPD), and the symptoms of the patients with mild asthma are often falsely thought to be caused by smoking. The objective of the study was to determine the prevalence of doctor-diagnosed asthma, asthmatic symptoms and doctor-diagnosed COPD in an adult population. The prevalence and relation to asthma of aspirin intolerance, nasal polyposis, allergic rhinitis and smoking habits were also examined. METHODS: Postal questionnaire survey of a population-based random sample (4300) of adult women and men aged 18-65 years served by the Paijat-Hame Central Hospital in southern Finland (a region with 208 000 inhabitants) was performed. RESULTS: The non-response-adjusted prevalence (Drane's linear method) of doctor-diagnosed asthma was 4.4% (95% CI: 3.3-5.5%) and of COPD 3.7% (95% CI: 2.7-4.8%). The prevalence of allergic rhinitis was 37.3% (95% CI: 33.3-41.2%), and of overall aspirin intolerance 5.7% (95% CI: 4.4 7.1%). The observed prevalence of aspirin intolerance causing shortness of breath or attacks of asthma was 1.2% and it was higher in patients with doctor-diagnosed asthma than without (8.8% versus 0.8%, relative risk [RR] = 11.4, P < 0.0001), and higher in those with allergic-like rhinitis than without (2.6% versus 0.3%, RR = 7.7, P < 0.0001). The prevalence of nasal polyposis was 4.3% (95% CI : 2.8 5.8%). CONCLUSIONS: The current prevalence of doctor-diagnosed asthma among adults is 4.4%, and allergic rhinitis, nasal polyposis and aspirin intolerance are associated with an increased risk of asthma. There is also association between aspirin-induced asthma and allergic-like rhinitis. PMID- 10480702 TI - Maternal infections in pregnancy and the development of asthma among offspring. AB - BACKGROUND: Previous studies have suggested that asthma phenotype could probably be programmed before birth. The current study examined the impact of maternal vaginitis and febrile infections during pregnancy on the subsequent development of asthma among children. METHODS: The analyses were based on 8088 children from the northern Finland birth cohort, 1985-1986. RESULTS: The prevalence of asthma at age 7 was 3.5%. Children had a higher risk of asthma if their mothers experienced vaginitis and febrile infections during pregnancy, odds ratio (OR) = 1.41, (95% CI: 1.08-1.84) and 1.65 (95% CI: 1.25-2.18), respectively, after adjusting for other covariates. There was a clear time trend in risk of childhood asthma corresponding to the timing of maternal febrile infections in pregnancy. The adjusted OR for the first, second and third trimesters were 2.08 (95% CI: 1.13-3.82), 1.73 (95% CI: 1.09-2.75) and 1.44 (95% CI: 0.97-2.15), respectively. Maternal history of allergic diseases, birthweight <2500 g and male gender also seemed to be risk factors for childhood asthma. CONCLUSIONS: Our results suggest that further investigation of the relation of maternal infections during pregnancy to asthma among children seems warranted. PMID- 10480703 TI - Risk of asthma in the general Spanish population attributable to specific immunoresponse. Spanish Group of the European Community Respiratory Health Survey. AB - BACKGROUND: Individuals with asthma are more atopic than the general population, but few reports have quantified this association, i.e. the proportion of asthma in the community due to specific immunoresponse. We aimed to determine the population attributable risk of asthma in the community due to atopy, and the quantity and quality of specific immunoresponses to common aeroallergens involved. METHODS: We used data from a cross-sectional study in a random sample of the general Spanish population, 20-44 years old, from five areas. In all, 1816 participants were given a symptoms questionnaire, a methacholine challenge and were assessed for atopy. Bronchial responsiveness (BR) was defined as a > or =20% fall in forced expiratory volume in one second (FEV1) after methacholine. Asthma was defined as symptomatic bronchial responsiveness. Atopy was assessed by measuring serum specific IgE or skin tests for sensitivity to Alternaria, birch, cat, Cladosporium, Dermatophagoides pteronyssinus, olive, Parietaria, ragweed or timothy grass. RESULTS: Sensitivity to any of the individual allergens tested significantly increased the risk of being asthmatic, at least twofold compared to non-atopic individuals. The population attributable risk of atopy in explaining asthma is 41.97% (95% CI: 29.2-60.3) when adjusting for area of residence, age, sex, and smoking. In symptomatic individuals, atopy was an independent factor in producing an early measurable PD20 (methacholine dose producing 20% fall in FEV1), especially in those atopic to two or more aeroallergens, and irrespective of the particular aeroallergen. CONCLUSIONS: Specific immunoresponse is strongly associated with asthma, but positivity to single allergens (quality) and the number of positive immunoresponses (quantity) do not change the per se risk of asthma in atopic individuals. PMID- 10480704 TI - Serum true insulin concentration and the risk of clinical non-insulin dependent diabetes during long-term follow-up. AB - BACKGROUND: There is considerable evidence that insulin resistance with compensatory hyperinsulinaemia is an early and modifiable defect in the pathogenesis of non-insulin dependent diabetes (NIDDM). Current data, however, are largely based on studies that have used insulin assays which cross-react with proinsulin and other insulin precursors. Using a specific assay, we have addressed the hypothesis that an elevation of serum true insulin concentration, reflecting insulin resistance, is an early event in the pathogenesis of NIDDM. METHODS: We have used a prospective cohort study design in which a group of 5550 non-diabetic men aged 40-59 years, from 18 British towns, have been followed for incident cases of physician-diagnosed NIDDM for an average of period of 14.8 years (range 13.5-15 years). We have estimated the incidence of physician diagnosed NIDDM by quintile of non-fasting serum true insulin concentration at entry into the study. RESULTS: There were 168 cases of clinically diagnosed NIDDM among the group of 5550 men during follow-up. Mean serum insulin at entry (geometric mean and 95% range, adjusted for time of sampling) was significantly higher in men who subsequently developed NIDDM than in the rest of the cohort, 19.5 mU/l (4.3-88.2) versus 12.2 mU/l (2.7-54.0), P < 0.0001. There was a highly significant linear trend of increasing risk of NIDDM by quintile of serum insulin which was not attenuated substantially after adjustment for age and body mass index (BMI) and additional lifestyle and biological factors associated with serum insulin and risk of NIDDM. However, in men with non-fasting serum glucose > or =6.1 mmol/l at baseline (80th percentile, n = 1125, 82 cases), the risk of NIDDM, adjusted for age and BMI, was higher in the first quintile of serum insulin than in all other quintiles. CONCLUSION: These findings are consistent with the hypothesis that the majority of cases of adult onset NIDDM in this population are characterized by the early development of insulin resistance with compensatory true hyperinsulinaemia. PMID- 10480705 TI - Thyroid nodules, thyroid function and dietary iodine in the Marshall islands. AB - BACKGROUND: Thyroid nodules have been found to be common in the population of the Marshall Islands. This has been attributed to potential exposure of radioiodines from the nuclear weapons tests on Bikini and Eniwetok between 1946 and 1958. METHODS: In order to get a full picture of thyroid pathology in the Marshallese population potentially exposed to radioactive fallout we performed a large thyroid screening programme using palpation, high resolution ultrasound and fine needle biopsies of palpable nodules. In addition, various parameters of thyroid function (free T3, free T4, thyroid stimulating hormone [TSH]) and anti-thyroid antibodies were examined in large proportions of the total population at risk. Since dietary iodine deficiency is an established risk factor for thyroid nodules, iodine concentration in urine samples of 362 adults and 119 children was measured as well as the iodine content of selected staple food products. RESULTS: The expected high prevalence of thyroid nodules was confirmed. There was no indication of an increased rate of impaired thyroid function in the Marshallese population. A moderate degree of iodine deficiency was found which may be responsible for some of the increased prevalence of thyroid nodules in the Marshallese population. CONCLUSIONS: Studies on the relationship between exposure to radioiodines and thyroid nodules need to take dietary iodine deficiency into account in the interpretation of findings. PMID- 10480707 TI - Mortality statistics in immigrant research: method for adjusting underestimation of mortality. AB - BACKGROUND: It is difficult to carry out fair comparisons of the mortality of different ethnic groups in a population in register-based studies because sizeable numbers of immigrants who subsequently leave their new homeland fail to register this fact with the national registration authorities. In this article we present a method which attempts to address these problems. METHODS: Age standardized mortality rates for native Swedes and immigrants in the age group 20 64 years were calculated for all individuals who either were included in the Swedish Population Censuses for 1985 or 1990, or who moved to Sweden during the period November 1990-1994. In order to define the population under scrutiny different sources of income are used as indicators of residence in the country. RESULTS: When an analysis is made of all nationally registered individuals, significantly reduced death rates are found among immigrants outside the north east of Europe compared to those for Swedish-born people. Extremely low death rates are found for those born in Turkey, Southern Europe, Latin America, Asia, and Africa and for those who are younger and without any income. When the income criterion is introduced, there is a change so that the earlier significantly reduced relative death risks for immigrants born outside the north-east of Europe for some subgroups are no longer significantly lowered. CONCLUSION: This study has important implications for the interpretation of every study of mortality among immigrants based on official mortality statistics. Using information about income as an indicator of residence in the country appears to be a method which can be pursued further in order to achieve a more accurate understanding of mortality among immigrant groups. PMID- 10480706 TI - The effect of recall on estimation of incidence rates for injury in Ghana. AB - BACKGROUND: Injury is a major public health problem in many developing countries. Due to limitations of vital registry and health service data, surveys are an important tool to obtain information about injury in these countries. The value of such surveys can be limited by incomplete recall. The most appropriate recall period to use in surveys on injury in developing countries has not been well addressed. METHODS: A household survey of injury in Ghana was conducted. Estimated annual non-fatal injury incidence rates were calculated for 12 recall periods (1-12 months prior to the interview, with each successively longer period including the preceding shorter periods). RESULTS: There was a notable decline in the estimated rate from 27.6 per 100 per year for a one-month recall period to 7.6 per 100 per year for a 12-month recall period (72% decline). The extent of this decline was not influenced by age, gender, rural versus urban location, nor by type of respondent (in-person versus proxy). Rate of decline was influenced by severity of injury. Injuries resulting in <7 days of disability showed an 86% decline in estimated rates from a one-month to a 12-month recall period, whereas injuries resulting in > or =30 days of disability showed minimal decline. CONCLUSIONS: In this setting, longer recall periods significantly underestimate the injury rate compared to shorter recall periods. Shorter recall periods (1-3 months) should be used when calculating the overall non-fatal injury incidence rate. However, longer recall periods (12 months) may be safely used to obtain information on the more severe, but less frequent, injuries. PMID- 10480708 TI - Case-case comparisons to study causation of common infectious diseases. AB - BACKGROUND: Analytical studies of reportable infectious diseases often use the small minority of cases detected through surveillance systems. For many diseases, notification of cases represents a non-random selection process. Apparent differences in exposure histories may be due to biases involved in the surveillance system selection of cases compared to randomly selected controls. In addition, differential recall between cases and controls may occur. One way to avoid these problems is to compare cases with another group of cases with a different disorder selected by a similar surveillance system, although this can introduce new biases. METHODS: In infectious diseases cases with the same disease can be divided into aetiologically meaningful subgroups by subtyping the pathogen. Exposure history can then be compared between these subgroups. RESULTS: Several biases are removed. The control group composed of other cases does not represent the exposure history of the study base but differs from it in a predictable and useful way. People considered as controls will have a higher incidence of general predisposing factors than the general population. Analysis is limited to factors associated with exposure to the infecting agent. CONCLUSIONS: Case-case comparison is a development of case-control methodology made possible by laboratory typing techniques. These comparisons allow a more restricted but more refined analysis of the association of some exposures with infection. Determination of how exposure to the infectious agent occurred is more efficient and unbiased than in standard case-control studies but general factors determining whether disease occurs after an infectious exposure can not be studied. PMID- 10480709 TI - Predictors and impact of losses to follow-up in an HIV-1 perinatal transmission cohort in Malawi. AB - BACKGROUND: Large simple trials which aim to study therapeutic interventions and epidemiological associations of human immunodeficiency virus (HIV) infection, including perinatal transmission, in Africa may have substantial rates of loss to follow-up. A better understanding of the characteristics and the impact of women and children lost to follow-up is needed. METHODS: We studied predictors and the impact of losses to follow-up of infants born in a large cohort of delivering women in urban Malawi. The cohort was established as part of a trial of vaginal cleansing with chlorhexidine during delivery to prevent mother-to-infant transmission of HIV. RESULTS: The HIV infection status could not be determined for 797 (36.9%) of 2156 infants born to HIV-infected mothers; 144 (6.7%) with missing status because of various sample problems and 653 (30.3%) because they never returned to the clinic. Notably, the observed rates of perinatal transmission were significantly lower in infants who returned later for determination of their infection status (odds ratio = 0.94 per month, P = 0.03), even though these infants must have had an additional risk of infection from breastfeeding. In multivariate models, infants of lower birthweight (P = 0.003) and, marginally, singletons (P = 0.09) were less likely to return for follow-up. The parents of infants lost to follow-up tended to be less educated (P < 0.001) and more likely to be in farming occupations, although one educated group, teachers and students, were also significantly less likely to return. Of these variables, infant birthweight, twins versus singletons, and maternal education were also associated with significant variation in the observed risk of perinatal transmission among infants of known HIV status. CONCLUSIONS: Several predictors of loss to follow-up were identified in this large HIV perinatal cohort. Losses to follow-up can impact the observed transmission rate and the risk associations in different studies. PMID- 10480710 TI - Hepatitis A incidence rate estimates from a pilot seroprevalence survey in Rio de Janeiro, Brazil. AB - BACKGROUND: To assess the impact of water sanitation and sewage disposal, part of a major environmental control programme in Rio de Janeiro, we carried out sero prevalence studies for Hepatitis A virus (HAV) in three micro-regions in Rio de Janeiro. Each region varied with regard to level of sanitation. We are interested in assessing the discriminating power of age-specific prevalence curves for HAV as a proxy for improvement in sanitation. These curves will serve as baseline information to future planned surveys as the sanitation programme progresses. METHODS: Incidence rate curves from prevalence data are estimated parametrically via a Weibull-like survival function, and non-parametrically via maximum likelihood and monotonic splines. Sera collected from children and adults in the three areas are used to detect antibodies against HAV through ELISA. RESULTS: We compare baseline incidence curves at the three sites estimated by the three methods. We observe a strong negative correlation between level of sanitation and incidence rates for HAV infection. Incidence estimates yielded by the parametric and non-parametric approaches tend to agree at early ages in the microregion showing the best level of sanitation and to increasingly disagree in the other two. CONCLUSION: Our results support the choice of HAV as a sentinel disease that is associated with level of sanitation. We also introduce monotonic splines as a novel non-parametric approach to estimate incidence from prevalence data. This approach outperforms current estimating procedures. PMID- 10480712 TI - A likelihood-based method of identifying contaminated lots of blood product. AB - BACKGROUND: In 1994 a small cluster of hepatitis-C cases in Rhesus-negative women in Ireland prompted a nationwide screening programme for hepatitis-C antibodies in all anti-D recipients. A total of 55 386 women presented for screening and a history of exposure to anti-D was sought from all those testing positive and a sample of those testing negative. The resulting data comprised 620 antibody positive and 1708 antibody-negative women with known exposure history, and interest was focused on using these data to estimate the infectivity of anti-D in the period 1970-1993. METHODS: Any exposure to anti-D provides an opportunity for infection, but the infection status at each exposure time is not observed. Instead, the available data from antibody testing only indicate whether at least one of the exposures resulted in infection. Using a simple Bernoulli model to describe the risk of infection in each year, the absence of information regarding which exposure(s) led to infection fits neatly into the framework of 'incomplete data'. Hence the expectation-maximization (EM) algorithm provides estimates of the infectiousness of anti-D in each of the 24 years studied. RESULTS: The analysis highlighted the 1977 anti-D as a source of infection, a fact which was confirmed by laboratory investigation. Other suspect batches were also identified, helping to direct the efforts of laboratory investigators. CONCLUSIONS: We have presented a method to estimate the risk of infection at each exposure time from multiple exposure data. The method can also be used to estimate transmission rates and the risk associated with different sources of infection in a range of infectious disease applications. PMID- 10480711 TI - Injections given in healthcare settings as a major source of acute hepatitis B in Moldova. AB - BACKGROUND: Reported rates of acute hepatitis B are high in many former Soviet Union republics and modes of transmission are not well defined. METHODS: Two case control studies were undertaken in Moldova to identify risk factors for acute hepatitis B in people aged 2-15 years (children) and > or =15 years (adults). Serologically confirmed acute hepatitis B cases occurring between 1 January 1994 and 30 August 30 1995, were matched on age, sex, and district of residence to three potential controls who were tested for hepatitis B markers to exclude the immune. Stratified odds ratios (SOR) were calculated using bivariate and multivariate methods. RESULTS: In multivariate analysis, compared with the 175 controls, the 70 adult cases (mean age 25 years, 66% male) were more likely to report receiving injections in the 6 months before illness during a dental visit (SOR = 21; 95% CI: 3.7-120), a hospital visit (SOR = 35; 95% CI: 7.2-170), or a visit to the polyclinic (SOR = 13; 95% CI: 2.4-74). Among children, receiving injections during a hospital visit (SOR = 5.2; 95% CI: 1.2-23) was the only exposure reported significantly more often by the 19 cases (mean age 8 years, 68% male) compared with the 81 controls. CONCLUSION: These results, along with reported unsafe injection practices in Moldova, suggest that injections are a major source of hepatitis B virus transmission and highlight the importance of proper infection-control procedures in preventing transmission of blood-borne infections. PMID- 10480713 TI - Longitudinal study of Plasmodium falciparum infection and immune responses in infants with or without the sickle cell trait. AB - BACKGROUND: Individuals may be homozygous (SS) or heterozygous (AS) sickle cell gene carriers or have normal adult haemoglobin (AA). Haemoglobin S could have a protective role against malaria but evidence is sparse and the operating mechanisms are poorly known. METHODS: We followed two cohorts of children. The first was enrolled at birth (156 newborn babies) and the second at 24-36 months old (84 children). Both cohorts were followed for 30 months; monthly for parasitological data and half yearly for immunological data. RESULTS: In the first cohort, 22%, and in the second 13% of children were AS. Whatever their age parasite prevalence rates were similar in AA and AS individuals. Mean parasite densities increased less rapidly with age in AS than in AA children, and were significantly lower in AS than in AA children >48 months old. The AA children tended to be more often admitted to hospital than AS children (22% versus 11%, NS). Both anti-Plasmodium falciparum and anti-Pfl55/RESA antibody rates increased more rapidly in AA than in AS children. Conversely, the prevalence rate of cellular responders to the Pfl55/RESA antigen was similar in AA and AS children during the first 2 years of life, then it was higher in AS than in AA children. CONCLUSIONS: Sickle cell trait related antimalarial protection varies with age. The role of the modifications of the specific immune response to P. falciparum in explaining the protection of AS children against malaria is discussed. PMID- 10480714 TI - Epilepsy and neurocysticercosis in an Andean community. AB - BACKGROUND: Taenia solium neurocysticercosis (NCC) has been documented as one of the major causes of epilepsy in developing countries. However, methodological limitations have hindered the evaluation of the epidemiological relationship between cysticercosis and epilepsy at the community level. METHODS: We used the WHO protocol for epidemiological evaluation of neurological disorders to conduct a door-to-door survey among 2723 residents of San Pablo del Lago, an Ecuadorean rural community in which T. solium taeniasis/cysticercosis was known to be endemic. The WHO protocol was complemented by neuroimaging and immunological tests to confirm the diagnosis of this infection. RESULTS: In all 31 people suffering from active epilepsy were detected (prevalence 11.4 per 1000, 95% CI:7.7-15.4); 26 agreed to undergo a computer tomography (CT) examination, and 28 agreed to have blood drawn for serodiagnosis. Fourteen of the 26 (53.8%) had CT changes compatible with NCC and six of the 28 (21.4%) tested positive in the enzyme-linked immunoelectro-transfer blot (EITB) assay. In a seizure-free random sample of this population, 17 of 118 (144 per 1000) subjects examined by CT and 10 out of 96 (104 per 1000) examined by EITB had evidence of this infection. The differences between the epilepsy group and the random sample of the population were statistically significant (OR = 6.93, 95% CI: 2.7-17.5, P < 0.001) for CT diagnosis, but not for EITB results (OR = 2.75, 95% CI: 0.8-7.1, P > 0.12, NS). CONCLUSIONS: These findings confirm that T. solium NCC is a significant cause of epilepsy at the community level in Andean villages of Ecuador. It is important to initiate effective public health interventions to eliminate this infection, which may be responsible for at least half of the cases of reported epilepsy in Ecuador. PMID- 10480715 TI - Epidemiology deserves better questionnaires. PMID- 10480716 TI - A reanalysis of data from regression models for ordinal responses'. PMID- 10480717 TI - ASP Annual Scientific Meeting 1998: will parasites outlive parasitologists? Australian Society for Parasitology. PMID- 10480718 TI - Eradication of Taenia solium cysticercosis: a role for vaccination of pigs. AB - Neurocysticercosis due to Taenia solium is an important cause of human morbidity and mortality, particularly in Latin America and parts of Africa and Asia. The disease has been recognised as potentially eradicable. Emphasis has been placed on control of the parasite through mass chemotherapy of human populations to remove tapeworm carriers. This strategy does not control the source of tapeworm infections, cysticercosis in pigs, and parasite transmission may continue due to incomplete chemotherapy coverage of human tapeworm carriers or because of immigration of tapeworm carriers into control areas. Exceptionally effective, practical vaccines have been developed against cysticercosis in sheep and cattle and a recent trial has proved recombinant antigens to be effective against Taenia solium cysticercosis in pigs. A new strategy for eradication of Taenia solium is proposed, based principally on a combined approach of chemotherapy of human tapeworm carriers and vaccination of all pigs at risk of infection. PMID- 10480719 TI - Funding options for research: facing the market as well as government. AB - Parasitology is a challenge. At one level, the structural and genetic complexities of parasites provide ample technical challenges in regard to an understanding of parasite variability and adaptability, epidemiological diversity, drug resistance, etc. The intricacies of host parasite relationships including the immunology of parasitism will continually surprise yet frustrate the vaccine developer and keep the bravest immunoparasitologist busy and creative for decades. As if the technical considerations were not challenging enough, we see difficulties arising in sustaining a research endeavour and preserving a critical mass of researchers through the generation of high-level, long-term funding support. Contributing to this situation is the fact that most parasitic diseases of major impact in humans are largely centred around the rural poor in tropical, less industrially-developed countries and therefore of little or of fickle interest to the strictly commercially oriented. Moreover, the focus in the rural industries has moved away from aspects of on-farm production with lower priority given to studies on even the 'economically-important' parasites of livestock. It is contended that this may change again with pressures and clear marketing advantages to preserving a 'clean and green' image for Australia's primary industries. Overall, the extraordinary technical and conceptual advances in recent times have been tempered by uncertainties in research funding and severe cuts from some traditional sources for both fundamental and strategic/applied research in Parasitology. Several have highlighted the fact that deliverables in terms of new methods of disease control have been sparse and some claims made in the past have certainly been exaggerated. Yet the prospects and achievements at the front end of the long R&D pathway have never been brighter. In this article we examine the merits of a 'portfolio approach' to generating research funds in Parasitology and Science and Technology in Australia more generally, with an emphasis on strategies that, through welding good science with clear, medium-term product objectives, increase research funding opportunities. PMID- 10480720 TI - Development of cysteine protease inhibitors as chemotherapy for parasitic diseases: insights on safety, target validation, and mechanism of action. AB - Cysteine proteases have been identified as promising targets for the development of antiparasitic chemotherapy. An attractive aspect of these enzymes is their widespread importance in both protozoan and helminth parasites of domestic animals and humans. Concerns about the ability to selectively inhibit parasite proteases without affecting host homologues have been addressed in recent studies of Trypanosoma cruzi and Plasmodium falciparum. Significant data on half-life, metabolism, pharmacokinetics and safety have been accumulated. Differential uptake of proteases by parasitic organisms versus host cells, and relatively less redundancy in parasite protease gene families, may be two factors which contribute to the successful treatment of animal models of infection. PMID- 10480721 TI - Aminopeptidases as potential targets for the control of the Australian sheep blowfly, Lucilia cuprina. AB - A series of experiments were carried out to investigate the role of proteinase enzymes in the growth of larvae of the sheep blowfly, Lucilia cuprina. First, instar larvae were incubated on an artificial growth media in the presence of various concentrations of inhibitors of all the major proteinase classes. Inhibitors of serine proteinases and aminopeptidases were found to cause significant growth inhibition and in some cases death of the larvae within 24 h, suggesting that these enzymes were the major classes involved in protein digestion in the gut of the insect. A second group of experiments analysed the effects of two inhibitors from the same or different proteinase classes in the growth media. Synergistic inhibition of larval growth was observed with the incorporation of inhibitors of serine proteinases and aminopeptidases. The results suggest that these classes of proteinases are both central to protein digestion in this insect, probably in the gut, and that the inhibition of both types of activity leads to an almost complete blockade of digestion. Testing in vivo gave similar results with infections on sheep skin inhibited by either serine proteinase or aminopeptidase enzyme inhibitors and the combination of both stopped the infection process. The role of aminopeptidases in larval metabolism and as potential targets for blowfly control agents is examined. PMID- 10480722 TI - Tick modulation of host immunity: an important factor in pathogen transmission. AB - Immunological interactions at the tick host interface involve innate and specific acquired host immune defenses and immunomodulatory countermeasures by the tick. Tick feeding stimulates host immune response pathways involving antigen presenting cells, cytokines, B-cells, T-cells, circulating and homocytotropic antibodies, granulocytes, and an array of biologically active molecules. In response to host immune defenses, tick-mediated host immunosuppressive countermeasures inhibit: host antibody responses; complement activation; T-cell proliferation; and cytokine elaboration by macrophages and Th1-lymphocytes. Immunosuppressive proteins identified in tick salivary glands and saliva have been partially characterised. Tick-induced host immunosuppression facilitates blood meal acquisition and is an important factor in the transmission/establishment of the tick-borne disease-causing agent, Borrelia burgdorferi. A novel strategy for control of tick-borne pathogens is proposed. PMID- 10480723 TI - Biochemical aspects of egg hatch in endo- and ectoparasites: potential for rational drug design. AB - Control of parasites through rational drug design requires a thorough understanding of the parasite's lifecycle encompassing the biochemical and physiological processes which contribute to normal parasite homeostasis. The hatching of parasite eggs for example, represents an important process in the development of a parasitic infection. Previous studies in helminths have indicated that secreted enzymes often facilitate successful endoparasite egg hatch. In contrast, there are relatively few examples demonstrating a role for secreted enzymes in the egg hatching process of insects. An analysis of this process in the ectoparasite Lucilia cuprina suggests a role for secreted enzymes in the hatching of sheep blowfly eggs. Characterisation of the proteases collected at the time of egg hatch indicates the presence of serine proteases. Further purification and characterisation of these proteases may enable the design of specific inhibitors to interfere with the egg hatch process and therefore provide a novel means of control. PMID- 10480724 TI - Do sheep regulate the size of their mallophagan louse populations? AB - Alternatives to chemicals for controlling parasites are required to minimise problems from resistance, residues in animal products and occupational exposure. Utilisation of host response to parasites through selection of resistant types or vaccination is an appealing option. To date most studies have been with haematophagous or invasive parasites which directly contact elements of the host immune system. Sheep lice (Bovicola ovis) feed superficially on the skin of sheep ingesting lipid, scurf, bacteria and loose stratum corneum squames. Evidence is presented that despite their surface feeding habit Bovicola ovis stimulate an immune response in sheep and that this response may play a part in regulating the size of louse populations. PMID- 10480725 TI - Parasitic helminths of the pig: factors influencing transmission and infection levels. AB - The occurrence of parasitic helminth species as well as infection intensities are markedly influenced by the type of swine production system used. The present review focusses mainly on the situation in temperate climate regions. Generally, over the past decades there has been a decrease in the number of worm species and worm loads in domestic pigs due to a gradual change from traditional to modern, intensive production systems. The reasons for some species being apparently more influenced by management changes than others are differences in the basic biological requirements of the pre-infective developmental stages, together with differences in transmission characteristics and immunogenicity of the different worm species. Control methods relevant for the different production systems are discussed. Outdoor rearing and organic pig production may in the future be confronted with serious problems because of particularly favourable conditions for helminth transmission. In addition, in organic farms preventive usage of anthelmintics is not permitted. PMID- 10480726 TI - The pathogenesis and control of diarrhoea and breech soiling in adult Merino sheep. AB - Diarrhoea and soiling of the breech with faeces ('winter scours') is a serious problem in adult Merino sheep grazing improved pastures in south-eastern Australia during winter and spring. This occurs even on farms where gastro intestinal nematodes are effectively controlled. It was shown that winter scours was associated with the ingestion of trichostrongylid larvae, and that host factors were important in determining susceptibility to this syndrome. No differences were detected in the protective immune response of affected and unaffected sheep to gut nematodes. However, affected sheep had a hypersensitive inflammatory reaction in the pylorus and upper jejunum, characterised by the infiltration of significantly more eosinophils and changed lymphocyte populations. The changes in the lymphocyte populations included a reduced number of CD8+ cells, increased CD4+:CD8+ T-cell ratio, and significantly reduced numbers of cells reacting to interferon-gamma. High doses of infective larvae (20,000/week of Ostertagia circumcincta and Trichostrongylus vitrinus) did not induce diarrhoea in sheep not susceptible to winter scours. In contrast, only low doses (2000/week) initiated scouring in sheep selected as being susceptible to winter scours. Therefore, even considerably improved worm control programmes, including the selection of sheep with increased resistance to gut nematodes, will not prevent winter scours. Rather, phenotypic culling and genetic selection, to remove sheep susceptible to the hypersensitivity inflammatory response, is proposed as the most suitable long-term control strategy. PMID- 10480727 TI - The functional importance of parasites in animal communities: many roles at many levels? AB - Past research on parasites and community ecology has focussed on two distinct levels of the overall community. First, it has been shown that parasites can have a role in structuring host communities. They can have differential effects on the different hosts that they exploit, they can directly debilitate a host that itself is a key structuring force in the community, or they can indirectly alter the phenotype of their host and change the importance of the host for the community. Second, certain parasite species can be important in shaping parasite communities. Dominant parasite species can directly compete with other parasite species inside the host and reduce their abundance to some extent, and parasites that alter host phenotype can indirectly make the host more or less suitable for other parasite species. The possibility that a parasite species simultaneously affects the structure of all levels of the overall community, i.e. the parasite community and the community of free-living animals, is never considered. Given the many direct and indirect ways in which a parasite species can modulate the abundance of other species, it is conceivable that some parasite species have functionally important roles in a community, and that their removal would change the relative composition of the whole community. An example from a soft-sediment intertidal community is used to illustrate how the subtle, indirect effects of a parasite species on non-host species can be very important to the structure of the overall community. Future community studies addressing the many potential influences of parasites will no doubt identify other functionally important parasite species that serve to maintain biodiversity. PMID- 10480728 TI - A new index of interactivity in parasite communities. AB - A new index of interactivity which allows objective evaluation and comparison of interactivity in communities between different host species is presented. The index is derived from the equations for species-accumulation curves generated using non-linear regression (with the Levenberg-Marquardt algorithm) of sample infracommunity richness data. It is advantageous in that it requires only presence/absence data to calculate, is applicable to all parasite taxa (including asexual species), is largely independent of sample size and allows objective comparison of parasite communities while correcting for differences in total richness. Iterative randomisation of infracommunity richness values to generate a mean value for the index avoids spurious results which may be generated by heterogeneity in infracommunity richness and the variation this produces in the non-linear regression results. PMID- 10480729 TI - Ecology of helminth communities in tropical Australian amphibians. AB - Less than 50% of Australian amphibians have been recorded as hosts for helminth parasites. Despite developments in parasite community ecology in amphibians elsewhere, Australia lags behind with only two publications on this subject. Reasons advanced for this are that much of the collecting and taxonomic studies were conducted earlier this century before more recent discoveries of host genera and species as well as species complexes in the amphibian fauna. Consequently, there is a need for re-collection of hosts and parasites, and taxonomic revision of the parasites. In addition, as shown in this study, the parasite fauna in Australian amphibians is depauperate. Composition of the parasite fauna was largely dependent on the ecological associations of the host animal species. Frogs were infected with few helminth species and these occurred at low intensity, indicating, as in Europe and North America, that a depauperate fauna is also characteristic of amphibians in tropical regions. PMID- 10480730 TI - Cytoadherence, pathogenesis and the infected red cell surface in Plasmodium falciparum. AB - The particular virulence of Plasmodium falciparum compared with the other malaria species which naturally infect humans is thought to be due to the way in which the parasite modifies the surface of the infected red cell. Approximately 16 hours into the asexual cycle, parasite encoded proteins appear on the red cell surface which mediate adherence to a variety of host tissues. Binding of infected red cells to vascular endothelium, a process which occurs in all infections, is thought to be an important factor in the pathogenesis of severe disease where concentration of organisms in particular organs such as the brain occurs. Binding to uninfected red cells to form erythrocyte rosettes, a property of some isolates, is linked to disease severity. Here we summarise the data on the molecular basis of these interactions on both the host and parasite surfaces and review the evidence for the involvement of particular receptors in specific disease syndromes. Finally we discuss the relevance of these data to the development of new treatments for malaria. PMID- 10480731 TI - The cytoadherence linked asexual gene family of Plasmodium falciparum: are there roles other than cytoadherence? AB - The binding of erythrocytes infected with Plasmodium falciparum to the endothelium lining the small blood vessels of the brain and other organs can mediate severe pathology. A region at the right end of chromosome 9 has been implicated in the binding of parasitised erythrocytes to the endothelial receptor CD36. A gene expressed in asexual erythrocytic stage parasites has been identified in this region and termed the cytoadherence linked asexual gene (clag). Antisense RNA production and targeted gene disruption of clag resulted in greatly reduced binding to CD36. Hybridisation to 3D7 chromosomes showed clag to be a part of a gene family of at least nine members. All members analysed so far have a conserved gene structure of at least nine exons, as well as putative transmembrane domains. The possible functions of the gene family are discussed. PMID- 10480732 TI - Transfection of the human malaria parasite Plasmodium falciparum. AB - In the past few years, methods have been developed which allow the introduction of exogenous DNA into the human malaria parasite Plasmodium falciparum. This important technical advance known as parasite transfection, provides powerful new tools to study the function of Plasmodium proteins and their roles in biology and disease. Already it has allowed the analysis of promoter function and has been successfully applied to establish the role of particular molecules and/or mutations in the biology of this parasite. This review summarises the current state of the technology and how it has been applied to dissect the function of the P. falciparum genome. PMID- 10480733 TI - Phylogenetic relationships of the genus Frenkelia: a review of its history and new knowledge gained from comparison of large subunit ribosomal ribonucleic acid gene sequences. AB - The different genera currently classified into the family Sarcocystidae include parasites which are of significant medical, veterinary and economic importance. The genus Sarcocystis is the largest within the family Sarcocystidae and consists of species which infect a broad range of animals including mammals, birds and reptiles. Frenkelia, another genus within this family, consists of parasites that use rodents as intermediate hosts and birds of prey as definitive hosts. Both genera follow an almost identical pattern of life cycle, and their life cycle stages are morphologically very similar. However, the relationship between the two genera remains unresolved because previous analyses of phenotypic characters and of small subunit ribosomal ribonucleic acid gene sequences have questioned the validity of the genus Frenkelia or the monophyly of the genus Sarcocystis if Frenkelia was recognised as a valid genus. We therefore subjected the large subunit ribosomal ribonucleic acid gene sequences of representative taxa in these genera to phylogenetic analyses to ascertain a definitive relationship between the two genera. The full length large subunit ribosomal ribonucleic acid gene sequences obtained were aligned using Clustal W and Dedicated Comparative Sequence Editor secondary structure alignments. The Dedicated Comparative Sequence Editor alignment was then split into two data sets, one including helical regions, and one including non-helical regions, in order to determine the more informative sites. Subsequently, all four alignment data sets were subjected to different tree-building algorithms. All of the analyses produced trees supporting the paraphyly of the genus Sarcocystis if Frenkelia was recognised as a valid genus and, thus, call for a revision of the current definition of these genera. However, an alternative, more parsimonious and more appropriate solution to the Sarcocystis/Frenkelia controversy is to synonymise the genus Frenkelia with the genus Sarcocystis. PMID- 10480734 TI - Genetics of host response to malaria. AB - A comprehension of the genetics of host resistance to malaria is essential to understanding the complex host/parasite interaction. Current research is directed towards the genetic dissection of both the murine and human host responses to the disease. Significant progress has been made towards the mapping of novel murine resistance loci. In addition, the role of the major histocompatibility complex in the host response has been examined in both animal models and human populations. Several large segregation analyses, association studies and, more recently, linkage analyses have been conducted in different African populations to examine the role of host genetics in both mild and severe malaria. The results of these studies have been collated within this review. The cloning of genes involved in malarial resistance will lead not only to a greater understanding of this complex disease but, potentially, to the development of effective medical intervention. PMID- 10480735 TI - Neuraminidase inhibitors as anti-influenza virus agents. AB - Influenza virus neuraminidase (NA) catalyses the cleavage of sialic acid residues terminally linked to glycoproteins and glycolipids and plays an important role in the replication of the virus. Recently, several potent NA inhibitors have been synthesized based on the rational design of mimicking the transition state of the sialic acid cleavage. Zanamivir and oseltamivir (GS 4104, the prodrug of GS 4071) have emerged as promising influenza NA inhibitors for the treatment and prophylaxis of human influenza virus infection. This review describes the recent work toward the discovery and development of influenza NA inhibitors. PMID- 10480736 TI - Approaches and strategies for the treatment of influenza virus infections. AB - Influenza A and B viruses belong to the Orthomyxoviridae family of viruses. These viruses are responsible for severe morbidity and significant excess mortality each year. Infection with influenza viruses usually leads to respiratory involvement and can result in pneumonia and secondary bacterial infections. Vaccine approaches to the prophylaxis of influenza virus infections have been problematic owing to the ability of these viruses to undergo antigenic shift by exchanging genomic segments or by undergoing antigenic drift, consisting of point mutations in the haemagglutinin (HA) and neuraminidase (NA) genes as a result of an error-prone viral polymerase. Historically, antiviral approaches for the therapy of both influenza A and B viruses have been largely unsuccessful until the elucidation of the X-ray crystallographic structure of the viral NA, which has permitted structure-based drug design of inhibitors of this enzyme. In addition, recent advances in the elucidation of the structure and complex function of influenza HA have resulted in the discovery of a number of diverse compounds that target this viral protein. This review article will focus largely on newer antiviral agents including those that inhibit the influenza virus NA and HA. Other novel approaches that have entered clinical trials or been considered for their clinical utility will be mentioned. PMID- 10480737 TI - Influence of treatment schedule and viral challenge dose on the in vivo influenza virus-inhibitory effects of the orally administered neuraminidase inhibitor GS 4104. AB - Experiments were done to determine how an alteration of the treatment schedule of 5 or 32 mg/kg/day per os (p.o.) doses of GS 4104 [the ethyl ester prodrug of the neuraminidase inhibitor (3R, 4R,5S)-4-acetamido-5-amino-3-(1-ethylpropoxy)-1 cylohexene-1 -carboxylic acid (GS 4071)] would affect influenza A (H1N1) virus infection in mice. Treatments with a low dose, one, two, three or four times daily, were highly inhibitory, unless therapy was terminated relatively early in the infection (days 2-3), in which case efficacy was curtailed. Single administrations at various times relative to virus exposure had essentially no effect. The 32 mg/kg/day dose was significantly inhibitory using all treatment schedules. These data indicated a requirement for the compound to be in the host when lung virus titres were reaching maximal levels and, for minimally effective doses, that at least continued daily therapy was needed to maintain adequate serum levels to achieve an appropriate antiviral effect. Twice daily p.o. treatment for 5 days with 20 mg/kg/day of GS 4104 totally prevented deaths in mice receiving high viral challenge doses that were sufficient to kill placebo treated controls in less than 5 days. Other parameters of antiviral efficacy (lung consolidation, arterial oxygen saturation, lung virus titres) were also markedly inhibited regardless of viral challenge doses. These data provide further insights into how the maximum therapeutic benefit can be derived from use of this orally effective influenza virus neuraminidase inhibitor. PMID- 10480738 TI - Anti-herpesvirus activities and cytotoxicities of 2-thiopyrimidine nucleoside analogues in vitro. AB - Twenty 2-thiopyrimidine nucleoside analogues were synthesized and examined for inhibitory activity against herpes simplex virus (HSV) type 1 and 2, varicella zoster virus (VZV), human cytomegalovirus (HCMV) and thymidine kinase-deficient HSV (HSV-TK-) replication in vitro. 2-thiouracil (thymine) arabinoside, 2'-deoxy 2-thiouridine (or 2-thiothymidine) and their 5-halogenated derivatives showed anti-HSV activity in both RPM18226 (human B-lymphoblastoid cells) and MRC-5 (human embryo lung cells). 2'-deoxy-5-halogenated-2-thiocytidines were also inhibitory against HSV, whereas 2-thiocytosine arabinoside and its derivatives were not inhibitory against HSV replication, except 5-bromo and 5-iodo congeners (TN-31, TN-32). Substitution of the halogen atom at the 5-position of the pyrimidine rings to an atom with a higher molecular weight increased anti-HSV and VZV activities, except for the anti-HSV activity of 2-thiouracil arabinosides. 2' deoxy-5-methyl-, and 2'-deoxy-5-iodo-2-thiouridines (TN-17, TN-44) showed the most potent anti-HSV activity, and 2'-deoxy-5-chloro- and 2'-deoxy-5-bromo-2 thiocytidines were potent inhibitors of VZV replication. However, none of the compounds inhibited HCMV and HSV-TK- replication. TN-31 and TN-32 were shown to inhibited HCMV and HSV-TK- as well as HSV and VZV replication. The cytotoxicity of the 2-thio-pyrimidine nucleoside analogues was less than that of the 2-oxy congeners of the compounds (5-iodo-2'-deoxyuridine, 5-iodo-2'-deoxycytidine, thymine arabinoside and cytosine arabinoside). The selectivity index of 2'-deoxy 5-iodo-2-thiouridine (TN-44) was higher than that of 5-iodo-deoxyuridine. TN-17 and TN-44 were not cytotoxic to resting or stimulated human peripheral blood mononuclear cells at 400 microM, although TN-32 was cytotoxic at a concentration of 20 microM. PMID- 10480740 TI - Transmission of religion and attitudes. PMID- 10480739 TI - Synthesis and biological activity of novel 1H,3H-thiazolo[3,4-a]benzimidazoles: non-nucleoside human immunodeficiency virus type 1 reverse transcriptase inhibitors. AB - Using a known human immunodeficiency virus type 1 (HIV-1) non-nucleoside reverse transcriptase inhibitor (NNRTI), 1-(2,6-difluorophenyl)-1H,3H-thiazolo[3,4 a]benzimidazole (TBZ NSC 625487) as the lead structure for drug design, a series of novel 1H,3H-thiazolo[3,4-a]benzimidazole derivatives substituted on the benzene-fused ring was prepared. Their in vitro anti-HIV-1 activity, as well as their inhibitory effects on the viral reverse transcriptase, were evaluated. The structure-activity relationships for these compounds are described and the results suggest that the apolar binding pocket of RT, into which the NNRTIs must fit, can accommodate only groups with a limited size and shape. PMID- 10480742 TI - Self-transcendence as a measure of spirituality in a sample of older Australian twins. AB - Measures of self-transcendence, physical health and psychological well-being were included in a self-report Health and Lifestyle questionnaire administered to Australian twins aged over 50 between 1993 and 1995. Self-transcendence appears to be higher among older Australian women than men, and was significantly associated with religious affiliation, marital status (in women) and age (in men). No strong correlations were observed between self-transcendence and any measure of psychological or physical health. Additive genetic effects were found to be important in influencing self-transcendence, with heritability estimates of 0.37 and 0.41 for men and women respectively, whilst shared environment effects were not found to be significant. Multivariate modelling of self-transcendence scores and self-reported church attendance behavior indicated substantially different etiologies for these variables, with implications for methods of investigation of religiosity and spirituality. PMID- 10480741 TI - Comparing the biological and cultural inheritance of personality and social attitudes in the Virginia 30,000 study of twins and their relatives. AB - Measures of four dimensions of personality (Psychoticism, Extraversion, Neuroticism, and Lie scores) and six aspects of social attitudes (to sex, taxation, militarism, politics, religion and a general conservatism scale) were obtained by mailed questionnaire from 29,691 US subjects including adult twins (n = 14,761) their parents (n = 2360), their spouses (n = 4391), siblings (n = 3184) and adult children (n = 4800). After correction for the average effects of age, sex and source of sample, familial correlations were computed for 80 distinct biological and social relationships. The data allow for the estimation of the additive and non-additive effects of genes, assortative mating, vertical cultural inheritance and other non-parental effects of the shared environment on differences in personality and social attitudes. The interaction of genetic and environmental effects with sex may also be analyzed. Model-fitting analyses show that personality and social attitude measures differ markedly in major features of family resemblance. Additive and dominant genetic effects contribute to differences in both personality and attitudes, but the effects of the family environment, including vertical cultural transmission from parent to child, are much more marked for social attitudes than for personality. There is substantial assortative mating for social attitudes and almost none for personality. The causes of family resemblance depend significantly on sex for almost every variable studied. These findings clarify and extend the more tentative findings derived from previous twin, family and adoption studies. PMID- 10480743 TI - Intrinsic and extrinsic religiousness: genetic and environmental influences and personality correlates. AB - This report presents findings for the Intrinsic (IR) and Extrinsic (ER) religiousness scales from the Minnesota Study of Twins Reared Apart. The scales were shown to be internally consistent, sufficiently distinct from the scales of the California Psychological Inventory and the Multidimensional Personality Questionnaire and unrelated to a number of measures of response style to justify treating them as distinct traits. The I scales also showed considerable evidence of construct validity in its correlations with religious fundamentalism and authoritarianism as assessed by the MMPI and Altemeyer's Right-Wing Authoritarianism scale. Data on IR and ER from 35 pairs of monozygotic twins reared apart (MZA) and 37 pairs of dizygotic twins reared apart (DZA) were fitted to a biometric model and demonstrated significant heritability (0.43 and 0.39), with a model containing genetic plus environmental factors fitting significantly better than a model containing only an environmental component. Twin similarity could not be explained by placement on a self-reported measure of family Moral Religious Emphasis as measured by the Family Environment Scale. PMID- 10480744 TI - Frequency of church attendance in Australia and the United States: models of family resemblance. AB - Data on frequency of church attendance have been obtained from separate cohorts of twins and their families from the USA and Australia (29,063 and 20,714 individuals from 5670 and 5615 families, respectively). The United States sample displayed considerably higher frequency of attendance at church services. Sources of family resemblance for this trait also differed between the Australian and US data, but both indicated significant additive genetic and shared environment effects on church attendance, with minor contributions from twin environment, assortative mating and parent-offspring environmental transmission. Principal differences between the populations were in greater maternal environmental effects in the US sample, as opposed to paternal effects in the Australian sample, and smaller shared environment effects observed for both women and men in the US cohort. PMID- 10480745 TI - Individual differences in adolescent religiosity in Finland: familial effects are modified by sex and region of residence. AB - Data from 16-year-old Finnish twin pairs were used to estimate familial effects on religiosity and the modification of those effects by sex and residential region. The sample of 2265 twin boys and 2521 twin girls formed 779 monozygotic and 1614 dizygotic pairs, 785 of the same sex and 829 of opposite sex. We compared religiosity scores of twins living in more rural and traditional northern Finland with those living in the more urban and secular southern region. Girls had higher religiosity scores than did boys, and twins living in northern Finland had higher religiosity scores than those resident in southern Finland. Correlations for monozygotic twins were slightly higher than those for dizygotic twins, and covariance modeling found modest heritability of religiosity [11% (95% CI 0-24) for girls; 22% (95% CI 6-38) for boys], and substantial shared environmental effects [60% (95% CI 49-69) and 45% (95% CI 31-57)] among girls and boys, respectively. The correlation between shared environmental effects in boys and girls was estimated to be 0.84 (95% CI 0.73-0.99). In analyses distinguishing region of residence, girls living in southern Finland were found to have significantly higher unshared environmental effects than girls in northern Finland, while boys living in the urban south appeared to have lower shared environmental effects, and higher additive genetic effects, than boys living in the rural north. PMID- 10480746 TI - A religious upbringing reduces the influence of genetic factors on disinhibition: evidence for interaction between genotype and environment on personality. AB - Information on personality, on anxiety and depression and on several aspects of religion was collected in 1974 Dutch families consisting of adolescent and young adult twins and their parents. Analyses of these data showed that differences between individuals in religious upbringing, in religious affiliation and in participation in church activities are not influenced by genetic factors. The familial resemblance for different aspects of religion is high, but can be explained entirely by environmental influences common to family members. Shared genes do not contribute to familial resemblances in religion. The absence of genetic influences on variation in several dimensions of religion is in contrast to findings of genetic influences on a large number of other traits that were studied in these twin families. Differences in religious background are associated with differences in personality, especially in Sensation Seeking. Subjects with a religious upbringing, who are currently religious and who engage in church activities score lower on the scales of the Sensation Seeking Questionnaire. The most pronounced effect is on the Disinhibition scale. The resemblances between twins for the Disinhibition scale differ according to their religious upbringing. Receiving a religious upbringing seems to reduce the influence of genetic factors on Disinhibition, especially in males. PMID- 10480747 TI - Religion and depression: a review of the literature. AB - We reviewed data from approximately 80 published and unpublished studies that examined the association of religious affiliation or involvement with depressive symptoms or depressive disorder. In these studies, religion was measured as religious affiliation; general religious involvement; organizational religious involvement; prayer or private religious involvement; religious salience and motivation; or religious beliefs. People from some religious affiliations appear to have an elevated risk for depressive symptoms and depressive disorder, and people with no religious affiliation are at an elevated risk in comparison with people who are religiously affiliated. People with high levels of general religious involvement, organizational religious involvement, religious salience, and intrinsic religious motivation are at reduced risk for depressive symptoms and depressive disorders. Private religious activity and particular religious beliefs appear to bear no reliable relationship with depression. People with high levels of extrinsic religious motivation are at increased risk for depressive symptoms. Although these associations tend to be consistent, they are modest and are substantially reduced in multivariate research. Longitudinal research is sparse, but suggests that some forms of religious involvement might exert a protective effect against the incidence and persistence of depressive symptoms or disorders. The existing research is sufficient to encourage further investigation of the associations of religion with depressive symptoms and disorder. Religion should be measured with higher methodological standards than those that have been accepted in survey research to date. PMID- 10480748 TI - Clarifying the relationship between religiosity and psychiatric illness: the impact of covariates and the specificity of buffering effects. AB - Previous analyses in a large population-based sample of female twins indicated that three dimensions of religiosity--personal devotion, personal conservatism and institutional conservatism--were, in different ways, significantly related to current depressive symptoms and substance use and lifetime psychiatric and substance use disorders. Furthermore, personal devotion, but neither personal conservatism nor institutional conservatism, buffered the depressogenic effects of stressful life events (SLEs). We here explore further these results, using linear, logistic and Cox regression models. Eight personality and six demographic variables had distinct patterns of association with the three dimensions. Personal devotion was positively associated with years of education, age, and optimism and negatively correlated with neuroticism. Personal conservatism was negatively associated with education, income, age, mastery and positively correlated with neuroticism. Institutional conservatism was negatively correlated with self-esteem and parental education. Covarying for these 14 variables produced little change in their association with psychiatric and substance use outcomes. The impact of the dimensions of religiosity differed as a function of the SLE category. High levels of both personal devotion and institutional conservatism protected against the depressogenic effects of death and personal illness. High levels of personal conservatism were associated with increased sensitivity to relationship problems. These results suggest that the association between religiosity and low risk for symptoms of depression and substance use may be in part causal. The relationship between dimensions of religiosity and response to SLEs is complex but probably of importance in clarifying the nature of the coping process. PMID- 10480749 TI - Resiliency factors protecting against teenage alcohol use and smoking: influences of religion, religious involvement and values, and ethnicity in the Missouri Adolescent Female Twin Study. AB - The objective of this study was to investigate the contribution of ethnicity (African American vs European/other ancestry), family religious affiliation, religious involvement, and religious values, to risk of alcohol and cigarette use in adolescent girls; and to estimate genetic and shared environmental effects on religious involvement and values. Telephone interviews were conducted with a sample of female like-sex twin pairs, aged 13-20 (n = 1687 pairs, including 220 minority pairs), as well as with one or both parents of twins aged 11-20 (n = 2111 families). These data, together with one-year follow-up twin questionnaire data, and two-year follow-up parent interview data, were used to compare ethnic differences. Proportional hazards regression models and genetic variance component models were fitted to the data. Despite higher levels of exposure to family, school and neighborhood environmental adversities, African American adolescents were less likely to become teenage drinkers or smokers. They showed greater religious involvement (frequency of attendance at religious services) and stronger religious values (eg belief in relying upon their religious beliefs to guide day-to-day living). Controlling for religious affiliation, involvement and values removed the ethnic difference in alcohol use, but had no effect on the difference in rates of smoking. Religious involvement and values exhibited high heritability in African Americans, but only modest heritability in EOAs. The strong protective effect of adolescent religious involvement and values, and its contribution to lower rates of African American alcohol use, was confirmed. We speculate about the possible association between high heritability of African American religious behavior and an accelerated maturation of religious values during adolescence. PMID- 10480750 TI - Adolescent religiousness and its influence on substance use: preliminary findings from the Mid-Atlantic School Age Twin Study. AB - Research has consistently shown that religiousness is associated with lower levels of alcohol and drug use, but little is known about the nature of adolescent religiousness or the mechanisms through which it influences problem behavior in this age group. This paper presents preliminary results from the Mid Atlantic School Age Twin Study, a prospective, population-based study of 6-18 year-old twins and their mothers. Factor analysis of a scale developed to characterize adolescent religiousness, the Religious Attitudes and Practices Inventory (RAPI), revealed three factors: theism, religious/spiritual practices, and peer religiousness. Twin correlations and univariate behavior-genetic models for these factors and a measure of belief that drug use is sinful reveal in 357 twin pairs that common environmental factors significantly influence these traits, but a minor influence of genetic factors could not be discounted. Correlations between the multiple factors of adolescent religiousness and substance use, comorbid problem behavior, mood disorders, and selected risk factors for substance involvement are also presented. Structural equation modeling illustrates that specific religious beliefs about the sinfulness of drugs and level of peer religiousness mediate the relationship between theistic beliefs and religious/spiritual practices on substance use. Limitations and future analyses are discussed. PMID- 10480751 TI - Religious attendance and frequency of alcohol use: same genes or same environments: a bivariate extended twin kinship model. AB - Religious attendance has been shown to correlate negatively with alcohol use. We investigated whether this relationship is driven by genetic or environmental factors. Data on frequency of church attendance and frequency of alcohol use were obtained from twins and their families in the Virginia 30,000 study. A comprehensive bivariate model of family resemblance was fitted to the data using Mx. This model is described in detail. Results indicate that genetic factors primarily account for the relationship between alcohol and church attendance in males, whilst shared environmental factors, including cultural transmission and genotype-environment covariance, are stronger determinants of this association in females. PMID- 10480752 TI - Vaccination against Lyme borreliosis. AB - Lyme borreliosis is a worldwide family of tick-borne infections caused by the spirochete Borrelia burgdorferi sensu lato. It is the most common tick-borne human infection in the Western world. There are several subgroups of the spirochete. Two monovalent vaccines against this infection have been presented in the USA, both of which use the borrelial outer surface protein A (OspA) as antigen. The first of these vaccines has been released for general use. A European polyvalent vaccine using the antigen OspC is undergoing clinical trial in the Aland Islands in Finland. Lately, another antigen group, decorin-binding proteins (Dbp), has been considered for immunization purposes. A European vaccine must be effective against several subgroups of the borrelia spirochete, and this complicates the situation compared with that in the USA, where one spirochete subspecies dominates the scene. PMID- 10480753 TI - Childhood weight and metabolic syndrome in adults. AB - Metabolic syndrome is a clustering of many insulin resistance-associated cardiovascular risk factors such as hypertension, hypertriglyceridaemia, low high density lipoprotein (HDL) cholesterol, abnormal glucose metabolism and hyperinsulinaemia. Furthermore, it is known that obesity is the most common clinical state characterized by insulin resistance. Central adiposity, in particular, has been shown to be the most distinctive feature of this syndrome. Some studies have also suggested that obesity per se would be necessary for the expression of metabolic defects associated with centrally distributed fat. It has been presented that undernutrition in utero might 'programme' blood pressure, insulin resistance, blood coagulation and cholesterol metabolism and would thus have a role in the aetiology of cardiovascular disease and type 2 diabetes in adult life. Some studies have also found associations between low birthweight and metabolic syndrome in adulthood. However, criticism on this hypothesis of fetal programming has recently been presented. It has been suggested that the origins of adulthood risk of cardiovascular disease and type 2 diabetes can be related to somatic growth as a child, not necessarily to intrauterine growth. In westernized countries, the relative proportion of underweight newborn children is decreasing, and thus considering entire populations low birthweight has lost its theoretical role in the aetiology of type 2 diabetes and cardiovascular disease. On the other hand, as obesity is known to be increasing in the industrialized countries among all age groups, the association between weight gain in childhood and metabolic syndrome in adulthood is more than noteworthy. Instead of undernutrition during pregnancy, sedentary lifestyle and lack of physical exercise pose a new threat. This results in an increased occurrence of overweight in childhood, which may be the first sign of insulin resistance and future metabolic syndrome. PMID- 10480754 TI - Autonomic mechanisms and sudden death after abrupt coronary occlusion. AB - In spite of recent advances in secondary prevention, sudden cardiac death has remained a major public health problem as the majority of fatalities occur in subjects without a history of severe heart disease. Abrupt rupture of a vulnerable plaque resulting in thrombotic occlusion of a coronary artery is a common cause of sudden death in this population. Coronary occlusion does not, however, invariably lead to sudden death but may cause acute myocardial infarction or exacerbation of chest pain. Extensive studies in experimental animals and increasing clinical evidence indicate that autonomic nervous activity has a significant role in modifying the clinical outcome. Sympathetic hyperactivity favours the genesis of life-threatening ventricular tachyarrhythmias while vagal activation exerts an antifibrillatory effect. Strong afferent stimuli from the ischaemic myocardium impair arterial baroreflex and may lead to dangerous haemodynamic instability. Studies with a human angioplasty model have shown that there is wide interindividual variation in the type and severity of autonomic reactions during the early phase of abrupt coronary occlusion, a critical period for out-of-hospital cardiac arrest. The site of the occlusion is not a significant determinant of the reactions, whereas the severity of a coronary stenosis, adaptation or ischaemic preconditioning, beta-blockade and gender seem to affect the autonomic reactions and occurrence of complex ventricular arrhythmias. Clinical and angiographic factors are, however, poor predictors of autonomic reactions in an individual patient. Recent studies have documented a hereditary component for autonomic function, and genetic factors may also modify the clinical manifestations of acute coronary occlusion. PMID- 10480755 TI - Chronic hypertension in pregnancy. AB - Pregnancies in women with chronic hypertension are at increased risk of superimposed pre-eclampsia, abruptio placentae, fetal growth retardation and prematurity. The frequencies of these complications are increased in those women who have high-risk chronic hypertension, ie severe hypertension or pre-existing cardiovascular or renal diseases, as well as in those with target organ damage. Such women should receive antihypertensive therapy and close management to improve maternal and fetal outcome. In women with low-risk chronic hypertension, antihypertensive treatments do not improve pregnancy outcome. Prophylactic low dose acetylsalicylic acid treatment does not reduce the frequency of superimposed pre-eclampsia nor does it improve perinatal outcome in these pregnancies. PMID- 10480756 TI - Salvage treatment against human immunodeficiency virus. AB - Despite dramatic declines in human immunodeficiency virus (HIV)-associated morbidity and mortality as a result of highly active antiretroviral combination therapies, including protease inhibitors, treatment failure occurs at such high rates as 20-50%. As drug regimens are very demanding, even short decreases of drug concentrations may trigger resistance. Viral loads can be decreased to very low concentrations, and there is no strict cut-off regarding the definition of treatment failure. Nevertheless, continuous detection of HIV of more than 50 copies per mL blood plasma is a predictor of increasing viral loads and of a suboptimal response to therapy. From a theoretical point of view, treatment changes should be made at low HIV RNA levels, but fewer options often dictate a more conservative approach. Drug susceptibility testing will be of increasing value, especially in patients experiencing drug failure for the first time. Success of salvage therapies is closely connected with the use of new compounds including new drug classes. As drugs susceptible to a multi-drug-resistant HIV are not yet available, regimens with more than three or even with five to nine drugs are used in clinical trials. Salvage therapies often fail in virological terms, ie in 50-80% of patients, depending primarily on the treatment history, but immunological and clinical stability can often be achieved. PMID- 10480757 TI - Heat shock proteins as cellular lifeguards. AB - Cells have developed complex ways to respond to various stresses. Interestingly, stresses such as heat, ischaemia and radiation can induce different cellular responses depending on their strength. While a mild stress induces a protective heat shock response, a more potent stress stimulus induces apoptosis and an even stronger one leads to necrosis. The heat shock or stress response, ie the synthesis of heat shock proteins (Hsps, stress proteins) in response to a mild stress, allows cells to adapt to gradual changes in their environment and to survive in otherwise lethal conditions. The ability of Hsps to protect cultured cells from both apoptosis and necrosis has been well demonstrated. Novel data suggest an important protective role for them also in vivo as they can protect heart and brain against ischaemia and lungs and liver against sepsis. Moreover, they can render tumours resistant to anticancer therapy. These and other cytoprotective effects of Hsps make them tempting targets for therapeutic interventions in several diseases. PMID- 10480758 TI - Clinical spectrum of food allergy in children in Australia and South-East Asia: identification and targets for treatment. AB - The prevalence of atopic diseases is increasing worldwide for reasons that are not clear. Food allergies are the earliest manifestations of atopy. This review defines the foods most commonly involved in allergic reactions and identifies an emerging group of syndromes in which food allergy is involved. A study of the frequency of food allergies in Australia and South-East Asia has recently shown that egg, cow's milk and peanut are the most common food allergens in Australia, but there were divergent results from different regions of South-East Asia. It is not clear whether the differences in reactivity to foods are due to genetic or cultural factors, but the findings raise the possibility that genetic susceptibility to food allergy may operate at the T-cell level modulated by the major histocompatibility complex. The Melbourne Milk Allergy Study defined a wide range of clinical symptoms and syndromes that could be reproduced by dietary challenge. A subsequent analysis of the infants with hypersensitivity to cow's milk and other multiple food proteins identified a new syndrome, multiple food protein intolerance of infancy. Food challenges demonstrated reactions developing slowly days after commencement of low-allergen soy formula or extensively hydrolysed formula. Follow-up at the age of 3 years showed that most children with this disorder tolerated most foods apart from cow's milk, egg and peanut. Atopic dermatitis affects about 18% of infants in the first 2 years of life. In a community-based study we have shown a very strong association (RR 3.5) between atopic dermatitis and infants with immunoglobulin E allergy to cow's milk, egg or peanut. Family studies on these infants have shown a link between atopic dermatitis and the genomic region 5q31 adjacent to the interleukin-4 gene cluster. Infantile colic (distress) affects 15-40% of infants in the first 4 months of life. Many theories of causation have been proposed, but a study from our centre showed that dietary modification, particularly that of breastfeeding mothers whose infants present with colic before the age of 6 weeks, alleviated symptoms. Colic associated with vomiting has been attributed to gastro oesophageal reflux (GOR). This has been considered primarily a motility disorder, but a secondary form resulting from food protein intolerance has been described recently. We have also recently identified a group of infants with distressed behaviour attributed to GOR who have failed to respond to H2-receptor antagonists, prokinetic agents and multiple formula changes. Symptoms resolved on commencement of an elemental amino acid-based formula. In two-thirds of the patients, symptoms relapsed when challenged with low-allergen soy formula or extensively hydrolysed formula. We propose that a period of food protein intolerance is a part of the normal development of the immune system as it encounters common dietary proteins in infancy and early childhood. Future targets for research are development of appropriate dietary and management strategies for these entities and identification of genetic markers for these disorders. PMID- 10480759 TI - Dietary fatty acids and allergy. AB - The increase in the prevalence of atopic diseases has recently been linked to altered consumption of polyunsaturated fatty acids (PUFAs). As typical Western diets contain almost 10 times more linoleic acid (18:2 omega-6) than alpha linolenic acid (18:3 omega-3), it is the metabolism of the former that predominates. Subsequently produced arachidonic acid-derived eicosanoids alter the balance of T-helper cells type 1 and type 2 thus favouring the production of immunoglobulin (Ig)E. In atopic subjects, the impact of this excessive eicosanoid production may be further strengthened as a result of changes in cyclic nucleotide metabolism exacerbated by substrate availability. Dietary omega-3 fatty acids can have marked influence on both specific and nonspecific immune responses in modifying eicosanoid production and replacing omega-6 fatty acids in cell membranes. Therefore, it is concluded that careful manipulation of dietary PUFAs may play a key role in the successful management of inflammation associated with atopic diseases. PMID- 10480760 TI - Healthy gut microflora and allergy: factors influencing development of the microbiota. AB - In humans, microbial colonization of the intestine begins just after birth. However, development of the normal flora is a gradual process, which is initially determined by factors such as composition of the maternal gut microflora, environment and possibly also by genetic aspects. A number of variables, such as the degree of hygiene, mode of delivery, use of antibiotics or other medication and a need for nursing in incubators, can all have a substantial effect on microbial colonization and development. Current knowledge on the significance and impact of such alterations on the health of the infant is poor. However, the essential role of the gut microflora in the development of the gut immune system indicates that a close relationship between allergic sensitization and the development of the intestinal microflora may occur in infancy. Intestinal micro organisms could down-regulate the allergic inflammation by counterbalancing type 2 T-helper cell responses and by enhancing antigen exclusion through an immunoglobulin (Ig)A response. The efficacy of probiotics (microbial food additions) in the management of food allergy has been demonstrated, and these data suggest that also prebiotics, food components that target certain indigenous gut bacteria, can possibly be used for this purpose. In conclusion, the developmental pattern of the normal gut microbiota in allergic infants poses an important research avenue, as the role of the gut microflora in the mechanisms of allergy, and thereby the possible targets for efficient bacteriotherapy, are currently undetermined. PMID- 10480761 TI - Benefits and risks of elimination diets. AB - Elimination diets have been used for the prevention or treatment of allergic disease with the diet of the pregnant or breastfeeding mother or the child, or both, being modulated as deemed appropriate. Evidence from studies published so far suggests that dietary restrictions are in fact effective only in the treatment of specific food allergies, not in allergy prevention. An elimination diet of a child or a breastfeeding mother entails a risk to normal nutrition and growth of the child. Although studies are lacking, dietary restrictions during lactation may well be harmful also to the mother's health. Substitution of nutritionally important foods and professional guidance are necessary for the successful treatment of food allergies. PMID- 10480762 TI - New functional foods in the treatment of food allergy. AB - Over the last two decades atopic diseases have increased in prevalence and severity in the industrialized countries. Food allergy is frequently the first manifestation of these disorders. Currently food allergy is mainly treated with elimination diets. However, this approach to control allergic inflammation by empirical elimination diets has not proved totally satisfactory. Applying therapeutic elimination diets in clinically documented allergy to a specific food has been shown to alleviate symptoms and reverse some disturbances of humoral and cell-mediated immune response. These diets are, however, associated with the risk of inadequate nutrition in infants with allergies to foods of vital importance. New approaches to the management of food allergy, such as for example immunotherapy for counteracting the hypersensitivity process and for potentiating the gut barrier mechanisms, are therefore needed. Diet remains an important element in this approach as sensitization to dietary antigens is frequently transient and is reversed to antigen-specific hyporesponsiveness. However, it represents an initial link in the development of more permanent sensitization to aeroallergens. PMID- 10480763 TI - Memory and metabolic control in children. PMID- 10480764 TI - Should high-dose vitamin E supplementation be recommended to diabetic patients? PMID- 10480765 TI - High-dose vitamin E supplementation normalizes retinal blood flow and creatinine clearance in patients with type 1 diabetes. AB - OBJECTIVE: To determine the effectiveness of vitamin E treatment in normalizing retinal blood flow and renal function in patients with <10 years of type 1 diabetes. RESEARCH DESIGN AND METHODS: An 8-month randomized double-masked placebo-controlled crossover trial evaluated 36 type 1 diabetic and 9 nondiabetic subjects. Subjects were randomly assigned to either 1,800 IU vitamin E/day or placebo for 4 months and followed, after treatment crossover, for a further 4 months. Retinal blood flow was measured using video fluorescein angiography, and renal function was assessed using normalized creatinine clearance from timed urine collections. RESULTS: After vitamin E treatment, serum levels of vitamin E were significantly elevated (P<0.01) in both type 1 diabetic and control patients. Hemoglobin A1c was not affected by vitamin E treatment. Diabetic patient baseline retinal blood flow (29.1+/-7.5 pixel2/s) was significantly (P = 0.030) decreased compared with that of nondiabetic subjects (35.2+/-7.2 pixel2/s). After vitamin E treatment, diabetic patient retinal blood flow (34.5+/ 7.8 pixel2/s) was significantly increased (P<0.001) and was comparable with that of nondiabetic subjects. Additionally, vitamin E treatment significantly (P = 0.039) normalized elevated baseline creatinine clearance in diabetic patients. CONCLUSIONS: Oral vitamin E treatment appears to be effective in normalizing retinal hemodynamic abnormalities and improving renal function in type 1 diabetic patients of short disease duration without inducing a significant change in glycemic control. This suggests that vitamin E supplementation may provide an additional benefit in reducing the risks for developing diabetic retinopathy or nephropathy. PMID- 10480766 TI - Use of a plastic insulin dosage guide to correct blood glucose levels out of the target range and for carbohydrate counting in subjects with type 1 diabetes. AB - OBJECTIVE: To improve glycemic control, a hand-held plastic Insulin Dosage Guide was developed to correct blood glucose levels outside of the target range. RESEARCH DESIGN AND METHODS: Protocol 1: Some 40 children (mean age 10.6+/-4.6 years) were randomly assigned for 3 months to use a written-on-paper algorithm or the Insulin Dosage Guide to correct abnormal blood glucose levels. Mean HbA1c and blood glucose levels and time to teach insulin dosage correction were compared. Protocol 2: The Insulin Dosage Guide was used by 83 subjects (mean age 11.4+/-4.3 years) for 1 year, and mean HbA1c levels, blood glucose levels, and number of consecutive high blood glucose values taken before and after the year were compared. Protocol 3: Some 20 patients (mean age 10.1+/-3.7 years) using rapid acting insulin and 64 patients (mean age 15.9+/-3.6 years) using an insulin pump and rapid-acting insulin used the Insulin Dosage Guide and had mean blood glucose levels, HbA1c, and percentage of blood glucose levels outside of the target range determined. RESULTS: Protocol 1: There was a significant reduction in mean HbA1c (P = 0.04) and blood glucose levels (P = 0.05) and in the time needed to teach how to correct blood glucose values using the Insulin Dosage Guide compared with the paper algorithm. Protocol 2: There was a decrease in mean HbA1c levels (P = 0.0001) and a decrease in the mean number of consecutive blood glucose levels (P = 0.001) over the 1-year time period. Protocol 3: With rapid-acting insulin, there was a significant increase in the percentage of blood glucose levels within the target range (1 month, P = 0.04; at 3 months, P = 0.03). With the insulin pump, there was a high rate (90%) of blood glucose levels in the target range during pump initiation when the Insulin Dosage Guide was used. CONCLUSIONS: This inexpensive hand-held plastic card, which is portable and easy to use, may help patients improve glycemia and successfully manage diabetes. PMID- 10480767 TI - Improved postprandial glycemic control during treatment with Humalog Mix25, a novel protamine-based insulin lispro formulation. Humalog Mix25 Study Group. AB - OBJECTIVE: Humalog Mix25 is a manufactured premixed insulin formulation containing insulin lispro and a novel insulin lispro-protamine formulation (NPL) in a ratio of 25:75%. The objective of this study was to compare Humalog Mix25 to human insulin 30/70 (30% regular insulin/70% NPH) with respect to glycemic control. RESEARCH DESIGN AND METHODS: Humalog Mix25 was compared with human insulin 30/70 in 89 individuals with type 2 diabetes during a 6-month randomized open-label two-period crossover study. Each insulin was administered twice daily, before the morning and evening meals. Information regarding self-monitored blood glucose (BG), hypoglycemic episodes (hypoglycemic signs or symptoms or BG < or = 3.0 mmol/l), insulin dose, and HbA1c was collected. RESULTS: Treatment with Humalog Mix25 resulted in better postprandial glycemic control after the morning and evening meals compared with treatment with human insulin 30/70. Overall glycemic control and the incidence of hypoglycemia were comparable between the treatments. CONCLUSIONS: In comparison to treatment with human insulin 30/70, twice daily administration of Humalog Mix25 resulted in improved postprandial glycemic control, similar overall glycemic control, and the convenience of dosing immediately before meals. PMID- 10480768 TI - Glucose intolerance and 23-year risk of coronary heart disease and total mortality: the Honolulu Heart Program. AB - OBJECTIVE: The associations between glucose intolerance measured at the study entry date and the 23-year incidence of coronary heart disease (CHD), CHD mortality, and total mortality were examined at the Honolulu Heart Program. RESEARCH DESIGN AND METHODS: This prospective study followed a cohort of 8,006 Japanese-American men who were 45-68 years old and living on the island of Oahu, HI, in 1965. Baseline glucose was measured in a nonfasting state 1 h after a 50-g glucose load. History and use of medication for diabetes was obtained during an interview. The cohort was divided into four categories of glucose tolerance: low normal, high-normal, asymptomatic hyperglycemia, and known diabetes. RESULTS: During the 23 years of follow-up, 864 incident cases of CHD, 384 deaths from CHD, and 2,166 total deaths occurred. The relative risks (RRs) were obtained using Cox proportional hazards modeling, with the low-normal category as a reference. The RRs were adjusted for age only, as well as for age, BMI, hypertension, cholesterol, triglycerides, smoking, alcohol, and a Japanese diet index. The age adjusted and risk factor-adjusted RRs for all outcomes were significant for the asymptomatic hyperglycemic and known diabetes groups (P<0.05). The age-adjusted RRs for CHD incidence and total mortality were marginally significant in the high normal group, but the RRs were not significant when adjusted for risk factors. CONCLUSIONS: These results suggest a dose-response relation of glucose intolerance at baseline with CHD incidence, CHD mortality, and total mortality, independent of other risk factors, in this cohort of middle-aged and older Japanese-American men. PMID- 10480769 TI - Weight change and duration of overweight and obesity in the incidence of type 2 diabetes. AB - OBJECTIVE: To examine the relationship between weight change and duration of overweight and obesity and the incidence of type 2 diabetes in a cohort of middle aged British men. RESEARCH DESIGN AND METHODS: We carried out a prospective study of cardiovascular disease in men aged 40-59 years at screening (1978-1980), drawn from one general practice in 24 British towns, who completed a postal questionnaire 5 years later (Q5) and for whom data on BMI at year 1 (Q1) and Q5 were available (n = 7,100). Men with diabetes at Q1 or Q5 and men with hyperglycemia at Q1 were excluded from the study (n = 184). The main outcome measure was type 2 diabetes (physician-diagnosed) during a mean follow-up period of 12 years starting at Q5 (1983-1985). RESULTS: In the 6,916 men with no history or evidence of diabetes, there were 237 incident cases of type 2 diabetes during the mean follow-up period of 12 years, a rate of 3.2/1,000 person-years. Substantial weight gain (>10%) was associated with a significant increase in risk of type 2 diabetes compared with that in men with stable weight (relative risk [RR] 1.61 [95% CI 1.01-2.56]) after adjustment for age, initial BMI, and other risk factors. Excluding men who developed diabetes within 4 years after the period of weight change increased the risk further (1.81 [1.09-3.00]). After adjustment and exclusion of men who developed diabetes early in the follow-up, weight loss (> or =4%) was associated with a reduction in the risk of type 2 diabetes, compared with that in the stable group, that reached marginal significance (0.65 [0.42-1.03], P = 0.07). A test for trend that fitted weight change as a continuous covariate showed the risk of diabetes to increase significantly from maximum weight loss to maximum weight gain (P = 0.0009). The lower risk associated with weight loss was seen in obese (> or =28 kg/m2) and nonobese subjects and in men with normal (<6.1 mmol/l) and high (> or =6.1 mmol/l) nonfasting blood glucose levels. Although not statistically significant, this is consistent with a benefit from weight loss. Risk of type 2 diabetes increased progressively and significantly with increasing levels of initial BMI and also with the duration of overweight and obesity (P<0.0001). CONCLUSIONS: This study confirms the critical importance of overweight and obesity, particularly of long duration, in the development of type 2 diabetes. The data support current public health recommendations to reduce the risk of type 2 diabetes by preventing weight gain in middle-aged men who are not overweight and by encouraging weight loss in overweight and obese men. PMID- 10480770 TI - Effects of repeated hypoglycemia on cognitive function: a psychometrically validated reanalysis of the Diabetes Control and Complications Trial data. AB - OBJECTIVE: To test the conclusion that there is no association between multiple episodes of severe hypoglycemia and cognitive decrements by reanalyzing the data from the Diabetes Control and Complications Trial (DCCT) with psychometrically validated cognitive factors and to conduct a novel analysis of the association between individual differences in baseline cognitive ability and episodes of severe hypoglycemia documented after baseline. RESEARCH DESIGN AND METHODS: The factor structure of cognitive ability in the neuropsychological data from the DCCT study was derived. Four cognitive factors (spatial ability, processing speed, memory, and verbal ability) were extracted. Changes in patients' cognitive scores for each year of follow-up were obtained, and paired comparisons of these change scores were performed between groups experiencing zero and five or more hypoglycemic episodes. The association between cognitive ability at baseline and number of subsequent episodes of severe hypoglycemia was also examined. RESULTS: Repeated episodes of hypoglycemia were found not to be associated with cognitive decline in any of the validated cognitive factors. No significant association was found between prospectively documented numbers of severe hypoglycemic episodes and baseline cognitive ability level. CONCLUSIONS: Repeated episodes of hypoglycemia were not related to cognitive decrement, and initial mental ability level was not associated with eventual numbers of hypoglycemic episodes in this group of patients. PMID- 10480771 TI - Association between stress and glycemic control in adults with type 1 (insulin dependent) diabetes. AB - OBJECTIVE: To examine the relationship between stressful life events and alterations in glycemic control in adults with diabetes. RESEARCH DESIGN AND METHODS: The occurrence of stressful experiences was recorded using the life Events and Difficulties Schedule of Brown and Harris in 55 adults with type 1 diabetes. The two most recent measures of glycemic control (HbA1c) were obtained from medical records, with poor glycemic control defined by the sample median (> or =7.7%). RESULTS: Subjects whose control deteriorated over time or who remained in poor glycemic control were significantly more likely to report severe personal stressors (SPS) in the month before HbA1c measurement, compared with subjects whose control remained fair or whose control improved (43 and 25% vs. 7 and 0%; P = 0.000). Subjects whose control remained fair or whose control improved were significantly more likely to report only positive life events during the same time period (80 and 11% vs. 0 and 0%, respectively; P = 0.000). Multiple regression analysis demonstrated that SPS, sex, and lack of further education were all significantly associated with either remaining in poor control or deterioration of control. CONCLUSIONS: The study has shown that recent severe stressors are associated with poorer glycemic control. Positive life events were associated with fair or improved glycemic control. This study has its limitations, and future studies should be prospective in design. While it is not always possible to avoid stress, learning to recognize and cope with stressors may help individuals with diabetes maintain good glycemic control and improve overall quality of life. PMID- 10480772 TI - Effects of maternal gestational diabetes on offspring adiposity at 4-7 years of age. AB - OBJECTIVE: The purpose of this study was to assess the long-term effects of maternal prenatal factors, including gestational diabetes mellitus (GDM), adiposity, and weight gain during pregnancy, on adiposity of offspring from 4 to 7 years of age. A second purpose was to investigate the relationships among childhood adiposity, blood pressure, and 2-h postprandial glucose level. RESEARCH DESIGN AND METHODS: Prospective observational study of four groups of children including large-for-gestational-age (LGA) offspring of mothers with gestational diabetes (OGDM); appropriate-for-gestational-age (AGA) OGDM; LGA control subjects; and AGA control subjects. Anthropometrics including skin-fold measurements, blood pressure measurements, and a 2-h postprandial glucose measurement were obtained at each visit. Repeated measures analysis of variance models were used to detect different patterns of longitudinal change among the groups. RESULTS: LGA OGDM were more likely to be heavier, have larger circumferences and skin-fold measurements, and have a higher BMI than AGA OGDM and control subjects, and these findings increased with increasing age. Blood pressures and postprandial glucose values were similar for OGDM and control subjects at 4-7 years. Multivariable analyses showed that infant BMI and maternal prepregnant BMI predicted 7-year BMI for OGDM, whereas for control subjects, maternal prepregnancy BMI and weight gain during pregnancy were positive predictors with a small negative contribution from birth BMI. CONCLUSIONS: We conclude that LGA OGDM have evidence of increasing body size and adiposity with increasing age and that maternal GDM and maternal prepregnant adiposity are significant predictors of their unique growth patterns. PMID- 10480773 TI - Early identification of diabetic foot ulcers that may require intervention using the micro lightguide spectrophotometer. AB - OBJECTIVE: Adequate tissue oxygenation is known to be essential for the healing of diabetic foot ulcers, but hypoxia has also been shown to be a potent stimulus for growth. There are no studies looking specifically at ulcer oxygen levels during the healing process. We measured the serial microvascular oxygen saturation (SaO2) of the foot ulcer, the ulcer margin, and a control site using the Erlangen micro lightguide spectrophotometer (EMPHO II; Bodenseewerk Geratetechnik, Erlangen, Germany) to study serial changes during healing. RESEARCH AND DESIGN METHODS: Studied over 9 months were 14 patients with neuropathy with a total of 24 foot ulcer sites. Of these patients, four (seven ulcers) had significant ischemia as determined by the ankle-brachial pressure index (ABPI) and transcutaneous oxygen tension. RESULTS: Of 21 ulcer sites with serial measurements, only 13 ulcers healed. In those ulcers, a significant reduction (P<0.05) in SaO2 occurred with healing. SaO2 dropped from 58% at initial presentation (mean area 2.6 cm2) to 47% at midsize (mean area 1.2 cm2 at 5.2 weeks) and finally reduced to 45% just before it healed. Similar trends were also seen around the margin of the ulcers (initial 49%, midsize 45%, and final 41%; P = 0.1). However, there were no such changes on the control sites (43, 40, and 40%; P = 0.5) or within the eight ulcers that did not heal (46, 42, and 53%; P = 0.2). CONCLUSIONS: Serial microvascular oxygen measurements may be used to identify at an early stage those ulcers that are unlikely to heal and, therefore, need surgical intervention. PMID- 10480775 TI - Use of automated telephone disease management calls in an ethnically diverse sample of low-income patients with diabetes. AB - OBJECTIVE: We examined whether low-income patients with diabetes were able and willing to use automated telephone disease management (ATDM) calls to provide health status information that could improve the quality of their care. RESEARCH DESIGN AND METHODS: A total of 252 adults with diabetes, 30 of whom spoke Spanish as their primary language, were enrolled at the time of clinic visits in a Department of Veterans Affairs health care system (n = 132) or a county health care system (n = 120). Patients received ATDM calls for 12 months and responded to queries using their touch-tone telephones. We examined 1) whether patients completed ATDM assessments consistently over the year and used the calls to report their self-monitored blood glucose (SMBG) levels, 2) the characteristics of patients most likely to use the system frequently, 3) whether patients responded consistently within ATDM assessments, and 4) whether ATDM assessments differentiated among groups of patients with different clinical profiles at baseline. RESULTS: Half of all patients completed at least 77% of their attempted assessments, and one-fourth completed at least 91%. Half of all patients reported SMBG levels during at least 86% of their assessments. Patients completed assessments and reported glucose levels consistently over the year. Health status indicators were the most important determinants of assessment completion rates, while socioeconomic factors were more strongly associated with patients' likelihood of reporting SMBG data during assessments. Patients' responses within assessments were consistent, and the information they provided during their initial assessments identified groups with poor glycemic control and other health problems. CONCLUSIONS: Most low-income patients with diabetes can and will use ATDM calls as part of their care. The information they provide is reliable and has clinical significance. ATDM calls could improve the information base for diabetes management while relieving some of the pressures of delivering diabetes care under cost constraints. PMID- 10480774 TI - Treatment of symptomatic diabetic polyneuropathy with the antioxidant alpha lipoic acid: a 7-month multicenter randomized controlled trial (ALADIN III Study). ALADIN III Study Group. Alpha-Lipoic Acid in Diabetic Neuropathy. AB - OBJECTIVE: To evaluate the efficacy and safety of alpha-lipoic acid given intravenously, followed by oral treatment in type 2 diabetic patients with symptomatic polyneuropathy. RESEARCH DESIGN AND METHODS: In a multicenter randomized double-blind placebo-controlled trial (Alpha-Lipoic Acid in Diabetic Neuropathy [ALADIN] III Study), 509 outpatients were randomly assigned to sequential treatment with 600 mg alpha-lipoic acid once daily intravenously for 3 weeks, followed by 600 mg alpha-lipoic acid three times a day orally for 6 months (A-A; n = 167); 600 mg alpha-lipoic acid once daily intravenously for 3 weeks, followed by placebo three times a day orally for 6 months (A-P; n = 174); and placebo once daily intravenously for 3 weeks, followed by placebo three times a day orally for 6 months (P-P; n = 168). Outcome measures included the Total Symptom Score (TSS) for neuropathic symptoms (pain, burning, paresthesias, and numbness) in the feet, and the Neuropathy Impairment Score (NIS). Data analysis was based on the intention to treat. RESULTS: No significant differences between the groups were noted for the demographic variables and the nerve function parameters at baseline. The TSS in the feet decreased from baseline to day 19 (median [range]) by -3.7 (-12.6 to 5.0) points in the group given alpha-lipoic acid intravenously and by -3.0 (-12.3 to 8.0) points in the placebo group (P = 0.447), but the area under curve on a daily basis was significantly smaller in the active as compared with the placebo group (85.6 [0-219] vs. 95.9 [5.5-220]); P = 0.033). After 7 months, the changes in the TSS from baseline were not significantly different between the three groups studied, which could be due to increasing intercenter variability in the TSS during the trial. The NIS decreased after 19 days by -4.34+/-0.35 points (mean +/- SEM) in A-A and A-P and -3.49+/ 0.58 points in P-P (P = 0.02 for alpha-lipoic acid versus placebo) and after 7 months by -5.82+/-0.73 points in A-A, -5.76+/-0.69 points in A-P, and -4.37+/ 0.83 points in P-P (P = 0.09 for A-A vs. P-P). The rates of adverse events were not different between the groups throughout the study. CONCLUSIONS: These findings indicate that a 3-week intravenous treatment with alpha-lipoic acid, followed by a 6-month oral treatment, had no effect on neuropathic symptoms distinguishable from placebo to a clinically meaningful degree, possibly due to increasing intercenter variability in symptom scoring during the study. However, this treatment was associated with a favorable effect on neuropathic deficits without causing significant adverse reactions. Long-term trials that focus on neuropathic deficits rather than symptoms as the primary criterion of efficacy are needed to see whether oral treatment with alpha-lipoic acid over several years may slow or reverse the progression of diabetic neuropathy. PMID- 10480776 TI - Dual energy X-ray absorptiometry assessment of fat mass distribution and its association with the insulin resistance syndrome. AB - OBJECTIVE: To determine which dual energy X-ray absorptiometry (DXA)-derived indices of fat mass distribution are the most informative to predict the various parameters of the metabolic syndrome. RESEARCH DESIGN AND METHODS: A total of 87 healthy men, 63 lean (% fat < or =26) and 24 obese (% fat >26), underwent DXA scanning to evaluate body composition with respect to the whole body and the trunk, leg, and abdominal regions from L1 to L4 and from L3 to L4. These regions were correlated with insulin sensitivity determined by the euglycemic hyperinsulinemic clamp, insulin area under the curve after oral glucose tolerance test (AUC I); triglyceride; total, HDL, and LDL cholesterol; free fatty acids; and blood pressure. The analyses were performed in all subjects, as well as in lean and obese groups separately. RESULTS: Among the various indices of body fat, DXA-determined adiposity in the abdominal cut at L1-4 level was the most predictive of the metabolic variables, showing significant relationships with glucose infusion rate ([GIR], mg kg(-1) lean body mass x min(-1)), triglyceride, and cholesterol, independent of total-body mass (r = -0.267, P<0.05; r = 0.316, P<0.005; and r = 0.319, P<0.005, respectively). Upon subanalysis, these correlations remained significant in lean men, whereas in obese men, only BMI and the amount of leg fat (negative relationship) showed significant correlations with triglyceride and cholesterol (r = 0.438, P<0.05; r = 0.458, P<0.05; r = 0.439, P<0.05; and r = -0.414, P<0.05, respectively). The results of a multiple regression analysis revealed that 47% of the variance in GIR among all study subjects was predicted by AUC I, fat L1-4, diastolic blood pressure (dBP), HDL, and triglyceride as independent variables. In the lean group, fat L1-4 alone accounted for 33% of the variance of GIR, whereas in obese men, AUC I and dBP explained 68% of the variance in GIR. CONCLUSIONS: The DXA technique applied for the evaluation of fat distribution can provide useful information regarding various aspects of the insulin resistance syndrome in healthy subjects. DXA can be a valid, accurate, relatively inexpensive, and safer alternative compared with other methods to investigate the role of abdominal body fat distribution on cardiovascular risk factors. PMID- 10480777 TI - Conventional versus intensive diabetes therapy in children with type 1 diabetes: effects on memory and motor speed. AB - OBJECTIVE: Severe hypoglycemia may impair medial temporal-mediated cognitive skills, such as the ability to recall past events explicitly (delayed declarative memory). The objective of this study was to determine whether delayed declarative memory deficits are present in a group of diabetic children with an increased risk of severe hypoglycemia. RESEARCH DESIGN AND METHODS: Nondiabetic children (n = 16) and children with type 1 diabetes who had been randomly assigned to either intensive (IT) (n = 13) or conventional (CT) (n = 12) diabetes therapy at the time of diagnosis participated in the study. All episodes of severe hypoglycemia were prospectively ascertained. All children were tested on memory tasks that have been closely linked to medial temporal functioning and on reaction time measures. RESULTS: Our results demonstrated that the IT group had a threefold higher rate of severe hypoglycemia, performed less accurately on a spatial declarative memory task, and performed more slowly, but not less accurately, on a pattern recognition task than did the CT group or control subjects. In addition, both groups of type 1 diabetic children were significantly impaired on a motor speed task compared with their nondiabetic peers. CONCLUSIONS: These results indicate a selective relative memory impairment associated with IT that is consistent with the effects of severe hypoglycemia and medial temporal damage or dysfunction. If larger prospective studies determine that severe hypoglycemia is the mediating factor for this memory impairment, extreme caution in imposing overly strict standards for glucose control in young patients with type 1 diabetes would be indicated because of the increased risk of hypoglycemia associated with IT regimens. PMID- 10480778 TI - Insulin sensitivity in African-American children with and without family history of type 2 diabetes. AB - OBJECTIVE: African-Americans are at increased risk for type 2 diabetes. We have previously demonstrated that African-American children are hyperinsulinemic and insulin resistant compared with their white American peers. The aim of the present investigation was to assess the impact of family history of type 2 diabetes on insulin sensitivity in African-American children. RESEARCH DESIGN AND METHODS: A total of 13 prepubertal healthy children with negative family history (FH-) and 9 with positive family history (FH+) of type 2 diabetes underwent a 3-h hyperinsulinemic (40 mU x m(-2) x min(-1))-euglycemic clamp study to assess insulin sensitivity. The groups were comparable for age, pubertal status, total body adiposity determined by dual-energy X-ray absorptiometry, abdominal adiposity assessed by computed tomography scan at the level of L4-5 lumbar vertebra, and physical fitness measured by maximal oxygen consumption (VO2max). RESULTS: The FH+, compared with the FH-, group had lower insulin-stimulated glucose disposal (10.9+/-1.2 vs. 14.2+/-0.9 mg x kg(-1) x min(-1), P = 0.035) and lower nonoxidative glucose disposal (5.7+/-0.8 vs. 8.3+/-0.6 mg x kg(-1) x min( 1), P = 0.015), with no differences in rates of glucose oxidation, fat oxidation, or insulin-mediated free fatty acid suppression. Fasting hepatic glucose production assessed with [6,6-2H2]glucose and basal rates of glucose and fat oxidation were not different between the two groups. CONCLUSIONS: These data suggest that in African-American children, family history of type 2 diabetes is a risk factor for insulin resistance. These children manifest important metabolic alterations, including impaired insulin-stimulated total and nonoxidative glucose disposal early in the first decade of life. We propose that this familial tendency, combined with environmental influences, could lead to type 2 diabetes decades later. PMID- 10480779 TI - Muscle fiber characteristics in postmenopausal women with normal or impaired glucose tolerance. AB - OBJECTIVE: Muscle fiber characteristics are altered in type 2 diabetes. We studied whether these alterations also exist in impaired glucose tolerance (IGT) and whether they are determinants of insulin sensitivity and glucose tolerance in postmenopausal women. RESEARCH DESIGN AND METHODS: Percutaneous muscle biopsies from the vastus lateralis muscle were obtained from 77 postmenopausal women aged 57-59 years: 50 women with normal glucose tolerance (NGT) and 27 with IGT. The IGT group had a reduced insulin sensitivity compared with the NGT group (euglycemic-hyperinsulinemic clamp) (P = 0.003). RESULTS: The groups did not differ in muscle fiber composition, as judged by the percentage of type I, IIa, or IIx fibers. In contrast, the IGT group had increased size of the IIa (mean +/ SD 3,776+/-987 vs. 3,078+/-862 microm2, P = 0.002) and IIx fibers (2,730+/-1,037 vs. 2,253+/-672 microm2, P = 0.017). There was a trend for the capillary diffusion areas (the muscle area supplied by each capillary) to be larger in the IGT group for the IIa (1,132+/-286 vs. 1,013+/-240 microm2, P = 0.061) and IIx fibers (1,020+/-246 vs. 906+/-240 microm2, P = 0.058). In the entire group, insulin sensitivity correlated with the size of the type IIa fibers (r = -0.28, P = 0.013), but not with the percentages of muscle fiber types. In a multiple regression, insulin sensitivity was determined by body fat content and HDL cholesterol level, while the size of the IIa fibers was not included in the model. Glucose tolerance was independently predicted by the number of capillaries/type I fiber, as well as by insulin sensitivity and triglyceride levels. CONCLUSIONS: We conclude that although muscle fiber composition is not altered, women with IGT have larger type IIa and IIx muscle fibers and a trend for increased capillary diffusion areas for these fibers, compared with women with NGT. In the entire group, insulin sensitivity was determined mainly by body fat content, while muscle fiber capillarization may be of importance for glucose tolerance. PMID- 10480780 TI - Plasma glucose within the normal range is not associated with carotid atherosclerosis: prospective results in subjects with normal glucose tolerance from the Bruneck Study. AB - OBJECTIVE: There is substantial evidence that glucose intolerance is associated with an increased risk of cardiovascular disease. However, it is not well established whether plasma glucose is independently related to atherosclerosis when glucose tolerance is normal and, if so, to which stage of the complex atherosclerotic process. RESEARCH DESIGN AND METHODS: We prospectively examined the status of carotid arteries in 625 subjects aged 40-79 years who were randomly selected from the general population and had normal glucose tolerance (according to World Health Organization criteria) both at baseline and at 5 years of follow up. All subjects had high-resolution echo-duplex evaluation of the common and internal carotid arteries (eight regions of interest on both sides) in 1990 and 1995 to detect the change in carotid status over time. The occurrence of new plaques in previously normal segments was termed "incident nonstenotic" or "early atherosclerosis," and the occurrence of stenosis in >40% of previously normal segments was termed "incident stenotic" or "advanced atherosclerosis." In addition, we evaluated the changes in the atherosclerosis score (the sum of all plaques) during the follow-up, and we measured intimal-medial thickening (IMT) in the common carotid artery in 1995. In all subjects, several candidate risk factors were assessed: sex, age, BMI, waist-to-hip ratio, glucose, HbA1c, insulin, urate, lipids, apolipoproteins A1 and B, blood pressure, lipoprotein(a), fibrinogen, antithrombin III, factor V Leiden mutation, ferritin, leukocyte count, smoking, alcohol intake, physical activity, and socioeconomic status. Fasting plasma glucose (FPG), plasma glucose 2 hr after the glucose load (2-h PG), and HbA1c concentrations in 1990 and 1995 were averaged in each subject to obtain an estimate of long-term glucose exposure of the arterial wall. RESULTS: Linear or logistical regression analyses indicated that neither baseline glucose and HbA1c levels nor mean FPG, mean 2-h PG, or mean HbA1c in 1990 and 1995 were independently related to IMT, a 5-year change in the atherosclerotic score, incident nonstenotic (early) atherosclerosis, or incident stenotic (advanced) atherosclerosis. Likewise, subjects with FPG levels above the median and subjects in the new category of "impaired fasting glucose" did not have an increased occurrence or progression of atherosclerosis. All results were consistent before and after adjustment for other vascular risk factors and possible confounders. CONCLUSIONS: These results suggest that plasma glucose levels within the normal range (<7.8 mg/dl both at FPG and 2-h PG) are not independently related to any stage of atherosclerosis. PMID- 10480781 TI - Chronic complications in patients with slowly progressing autoimmune type 1 diabetes (LADA). AB - OBJECTIVE: To study the prevalence of chronic diabetic complications in patients with the slowly progressing autoimmune form of type 1 diabetes, also referred to as latent autoimmune diabetes in adults (LADA). RESEARCH DESIGN AND METHODS: We evaluated factors associated with chronic diabetic complications in 59 patients with GAD antibodies (GADAs) and age at onset of diabetes >35 years and in 59 GADA negative type 2 diabetic patients. The prevalence of chronic complications was further compared with the prevalence in 111 type 1 diabetic patients. RESULTS: The LADA patients had lower BMI (P = 0.04), waist-to-hip ratio (P = 0.02 for men and P = 0.03 for women), and fasting C-peptide concentrations (P<0.001) higher HDL2 concentrations (P = 0.04), and less hypertension (58 vs. 75%, P = 0.05) than the type 2 diabetic patients. These differences were even more marked in patients with short disease duration. The prevalence of retinopathy (51 vs. 56%), neuropathy (29 vs. 27%), and microalbuminuria (27 vs. 29%) did not differ between the groups. The type 1 diabetic patients had lower prevalence of neuropathy (13%, P = 0.02) and higher prevalence of retinopathy (76%, P = 0.002) compared with the other groups. Neither the prevalence of coronary heart disease (CHD) (56 vs. 58%) nor cardiovascular mortality (7.4 vs. 12.4%, P = 0.2) significantly differed between the LADA and type 2 diabetic patients. In a multiple logistic regression analysis, glycemic control was associated with CHD (P = 0.02) in the LADA group but not in the type 2 diabetic group. CONCLUSIONS: Glycemic control is a stronger risk factor for cardiovascular disease in LADA patients than in patients with type 2 diabetes. This could be related to the lower prevalence of the metabolic syndrome seen in the former. PMID- 10480782 TI - Consensus Development Conference on Diabetic Foot Wound Care: 7-8 April 1999, Boston, Massachusetts. American Diabetes Association. PMID- 10480783 TI - Reproducibility of HbA1c in a healthy adult population: the Telecom Study. PMID- 10480784 TI - The European Association for the Study of Diabetes Annual Meeting, 1998: complications of diabetes. PMID- 10480785 TI - Improved stability of insulin delivery from implanted pumps using a new preparation process for infused insulin. PMID- 10480786 TI - Intensive insulin therapy compared with conventional insulin therapy does not reduce depressive symptoms in parents of children with type 1 diabetes. PMID- 10480787 TI - Sustained improvement of glycemic control by insulin treatment after 9 years in patients with type 2 diabetes and secondary failure. PMID- 10480789 TI - Hypoglycemia due to folk medicine. PMID- 10480788 TI - Late-onset lipoatrophic diabetes. Phenotypic and genotypic familial studies and effect of treatment with metformin and lispro insulin analog. PMID- 10480790 TI - Role of early fundoscopy for diagnosis of Wolfram syndrome in type 1 diabetic patients. PMID- 10480791 TI - Partial Wolfram syndrome (DIDMOAD): two new patients in a family. Diabetes insipidus, diabetes mellitus, optic atrophy, and deafness. PMID- 10480792 TI - Are insulin-treated type 2 diabetic subjects at higher risk for foot ulcers? PMID- 10480793 TI - Hyperglycemic hyperosmolar nonketotic syndrome as initial presentation of type 2 diabetes in a young cocaine abuser. PMID- 10480794 TI - Serum gamma-glutamyltransferase and/or serum cholinesterase as markers of the metabolic syndrome. PMID- 10480795 TI - A Japanese case of liver dysfunction after 19 months of troglitazone treatment. PMID- 10480796 TI - Postprandial oxidized lipids in diabetic patients: which role for postprandial hyperglycemia? PMID- 10480797 TI - Fasting blood glucose and risk of cardiovascular disease. PMID- 10480798 TI - Diagnosis of cardiac autonomic neuropathy in diabetic patients. PMID- 10480799 TI - Transitorine, a new quinoline alkaloid from ephedra transitoria PMID- 10480800 TI - Trans-atlantic conference on clinical trial guidelines in PAOD (Peripheral arterial occlusive disease) clinical trial methodology. PMID- 10480801 TI - Environmental management for vector control. Is it worth a dam if it worsens malaria? PMID- 10480802 TI - Can it work? Does it work? Is it worth it? The testing of healthcareinterventions is evolving. PMID- 10480803 TI - Accepting commercial sponsorship. Disclosure helps--but is not a panacea. PMID- 10480804 TI - Legal safeguards for the audit process. Are essential for effective clinical governance. PMID- 10480805 TI - Zanamivir for influenza: a public health perspective. Its use will require careful management by GPs. PMID- 10480806 TI - AMA backs health reform legislation. PMID- 10480807 TI - NIH website of original research to start in January. PMID- 10480809 TI - Medical advice in press is often unsuitable, sometimes dangerous. PMID- 10480808 TI - Baby to be tested for HIV against parents' wishes. PMID- 10480811 TI - Anti-flu drug may not reduce death rate. PMID- 10480812 TI - China moves to tackle iodine deficiency. PMID- 10480813 TI - UK watchdog issues guidelines to combat medical research fraud. PMID- 10480814 TI - Storm rages over revisions to Helsinki Declaration. PMID- 10480815 TI - ACE inhibitor reduces cardiovascular events by 22%. PMID- 10480817 TI - New EU health commissioner airs his priorities. PMID- 10480816 TI - Project makes emergency pill more available. PMID- 10480818 TI - Commons committee criticises Guy's fiasco. PMID- 10480819 TI - Apo E4 gene linked to breast cancer. PMID- 10480820 TI - Incidence of malaria among children living near dams in northern Ethiopia: community based incidence survey. AB - OBJECTIVE: To assess the impact of construction of microdams on the incidence of malaria in nearby communities in terms of possibly increasing peak incidence and prolonging transmission. DESIGN: Four quarterly cycles of malaria incidence surveys, each taking 30 days, undertaken in eight at risk communities close to dams paired with eight control villages at similar altitudes but beyond flight range of mosquitoes. SETTING: Tigray region in northern Ethiopia at altitudes of 1800 to 2225 m. SUBJECTS: About 7000 children under 10 years living in villages within 3 km of microdams and in control villages 8-10 km distant. MAIN OUTCOME MEASURES: Incidence of malaria in both communities. RESULTS: Overall incidence of malaria for the villages close to dams was 14.0 episodes/1000 child months at risk compared with 1.9 in the control villages-a sevenfold ratio. Incidence was significantly higher in both communities at altitudes below 1900 m. CONCLUSIONS: There is a need for attention to be given to health issues in the implementation of ecological and environmental development programmes, specifically for appropriate malaria control measures to counteract the increased risks near these dams. PMID- 10480821 TI - Cirrhosis mortality and per capita consumption of distilled spirits, United States, 1949-94: trend analysis. AB - OBJECTIVE: To describe, evaluate, and suggest interpretations for an observed aggregate level relation between trends in mortality from cirrhosis and per capita consumption of distilled spirits in the United States. DESIGN: Trend analysis using data on US cirrhosis mortality and per capita alcohol consumption. RESULTS: There is a consistent long term trend relation between mortality from cirrhosis and per capita consumption of distilled spirits in the United States from 1949 to 1994. Two instances of comparatively sharp drops in the consumption of spirits earlier in the 1940s generated mixed results in predicting changes in cirrhosis mortality. CONCLUSIONS: An aggregate level relation between trends in long term cirrhosis mortality and the consumption of spirits falls considerably short of establishing a direct causal link between the two for individuals. Moreover, two sharp drops in the consumption of spirits generated only mixed results with respect to the short term trend in cirrhosis. Nevertheless, the observed relation between the consumption of spirits and cirrhosis mortality merits further investigation. PMID- 10480822 TI - What is meant by intention to treat analysis? Survey of published randomised controlled trials. AB - OBJECTIVES: To assess the methodological quality of intention to treat analysis as reported in randomised controlled trials in four large medical journals. DESIGN: Survey of all reports of randomised controlled trials published in 1997 in the BMJ, Lancet, JAMA, and New England Journal of Medicine. MAIN OUTCOME MEASURES: Methods of dealing with deviations from random allocation and missing data. RESULTS: 119 (48%) of the reports mentioned intention to treat analysis. Of these, 12 excluded any patients who did not start the allocated intervention and three did not analyse all randomised subjects as allocated. Five reports explicitly stated that there were no deviations from random allocation. The remaining 99 reports seemed to analyse according to random allocation, but only 34 of these explicitly stated this. 89 (75%) trials had some missing data on the primary outcome variable. The methods used to deal with this were generally inadequate, potentially leading to a biased treatment effect. 29 (24%) trials had more than 10% of responses missing for the primary outcome, the methods of handling the missing responses were similar in this subset. CONCLUSIONS: The intention to treat approach is often inadequately described and inadequately applied. Authors should explicitly describe the handling of deviations from randomised allocation and missing responses and discuss the potential effect of any missing response. Readers should critically assess the validity of reported intention to treat analyses. PMID- 10480823 TI - Psychiatric disorder among children at time of entering local authority care: questionnaire survey. PMID- 10480824 TI - Multifaceted shared care intervention for late life depression in residential care: randomised controlled trial. AB - OBJECTIVE: To evaluate the effectiveness of a population based, multifaceted shared care intervention for late life depression in residential care. DESIGN: Randomised controlled trial, with control and intervention groups studied one after the other and blind follow up after 9.5 months. SETTING: Population of residential facility in Sydney living in self care units and hostels. PARTICIPANTS: 220 depressed residents aged >/=65 without severe cognitive impairment. INTERVENTION: The shared care intervention included: (a) multidisciplinary consultation and collaboration, (b) training of general practitioners and carers in detection and management of depression, and (c) depression related health education and activity programmes for residents. The control group received routine care. MAIN OUTCOME MEASURE: Geriatric depression scale. RESULTS: Intention to treat analysis was used. There was significantly more movement to "less depressed" levels of depression at follow up in the intervention than control group (Mantel-Haenszel stratification test, P=0.0125). Multiple linear regression analysis found a significant intervention effect after controlling for possible confounders, with the intervention group showing an average improvement of 1.87 points on the geriatric depression scale compared with the control group (95% confidence interval 0.76 to 2.97, P=0.0011). CONCLUSIONS: The outcome of depression among elderly people in residential care can be improved by multidisciplinary collaboration, by enhancing the clinical skills of general practitioners and care staff, and by providing depression related health education and activity programmes for residents. PMID- 10480825 TI - Improving the health behaviours of elderly people: randomised controlled trial of a general practice education programme. AB - OBJECTIVES: To establish the effect of an educational intervention for general practitioners on the health behaviours and wellbeing of elderly patients. DESIGN: Randomised controlled trial with 1 year follow up. SETTING: Metropolitan general practices in Melbourne, Australia. SUBJECTS: 42 general practitioners and 267 of their patients aged over 65 years. INTERVENTION: Educational and clinical practice audit programme for general practitioners on health promotion for elderly people. MAIN OUTCOME MEASURES: Patients' physical activity, functional status, self rated health, immunisation status, social contacts, psychological wellbeing, drug usage, and rate of influenza vaccination. Primary efficacy variables were changes in outcome measures over 1 year period. RESULTS: Patients in the intervention group had increased (a) walking by an average of 88 minutes per fortnight, (b) frequency of pleasurable activities, and (c) self rated health compared with the control group. No change was seen in drug usage, rate of influenza vaccination, functional status, or psychological wellbeing as a result of the intervention. Extrapolations of the known effect of these changes in behaviour suggest mortality could be reduced by 22% if activity was sustained for 5 years. CONCLUSIONS: Education of the general practitioners had a positive effect on health outcomes of their elderly patients. General practitioners may have considerable public health impact in promotion of health for elderly patients. PMID- 10480827 TI - The doctor's fault. PMID- 10480826 TI - Five year follow up of patients at high cardiovascular risk who took part in randomised controlled trial of health promotion. PMID- 10480829 TI - What is complementary medicine? PMID- 10480828 TI - Recent advances. Gynaecology. PMID- 10480830 TI - Always take a proper drug history. PMID- 10480831 TI - For debate. The terminology used for "cot deaths". PMID- 10480832 TI - New global Health for All targets. PMID- 10480833 TI - How to randomise. PMID- 10480834 TI - Generalisation of salt infusions. PMID- 10480835 TI - Guidelines dismay disabled people unable to speak for themselves. PMID- 10480836 TI - Measuring outcomes in economic evaluations. This economics note is misleading. PMID- 10480837 TI - Treatment of oral cancer. Radiotherapy may be as effective as surgery. PMID- 10480838 TI - Concerns about privacy in research may be exaggerated. PMID- 10480839 TI - Minimising the impact of visual impairment. From October this will have to be done to conform to the law. PMID- 10480840 TI - Antithrombotic treatment for atrial fibrillation. Patients must be told full details of risks of treatment. PMID- 10480841 TI - Rationing health care in New Zealand. Explicit rationing needs more debate. PMID- 10480842 TI - Greenwich asthma study. PMID- 10480843 TI - Psychotherapy for severe personality disorder. Evolution is part of the therapeutic process of therapeutic communities. PMID- 10480844 TI - Are postal prompts really ineffective? PMID- 10480845 TI - Factors that might have eased principals' entry into practice. PMID- 10480846 TI - Effectiveness of planned short hospital stays for mental health care. Older studies' definitions of lengths of stay are now outdated. PMID- 10480847 TI - Moving beyond journals. Print journals perform important functions. PMID- 10480848 TI - Patrick Forsyth Aitken Crawshaw. PMID- 10480849 TI - The Symbolic Species: The Co-Evolution of Language and the Human Brain. PMID- 10480850 TI - HIV and AIDS: Testing, Screening, and Confidentiality. PMID- 10480851 TI - An end to health scares? PMID- 10480852 TI - Sudden infant death syndrome. PMID- 10480854 TI - Spaced out. PMID- 10480853 TI - More doctors would aid communication. PMID- 10480855 TI - An ABC of complementary medicine: a new dawn. PMID- 10480856 TI - Malaria incidence is increased around irrigation projects. PMID- 10480857 TI - Cirrhosis mortality is linked with consumption of spirits. PMID- 10480858 TI - Intention to treat analysis is often inadequate. PMID- 10480860 TI - Multifaceted shared care improves outcomes for elderly depressed people. PMID- 10480859 TI - Children entering care have serious psychiatric disorders. PMID- 10480861 TI - Education of general practitioners improves the health of their elderly patients. PMID- 10480862 TI - Raman spectroscopy, the sleeping giant in structural biology, awakes. PMID- 10480863 TI - Acetylation of human hemoglobin by methyl acetylphosphate. Evidence of broad regio-selectivity revealed by NMR studies. AB - The development of chemical modification agents that reduce the tendency of sickle hemoglobin (HbS) to aggregate represents an important chemotherapeutic goal. Methyl acetylphosphate (MAP) has been reported to bind to the 2,3 diphosphoglycerate (2,3-DPG) binding site of hemoglobin, where it selectively acetylates residues, resulting in increased solubility of HbS. We have prepared [1-(13)C]MAP and evaluated the adduct formation with hemoglobin using (1)H-(13)C HMQC and HSQC NMR studies. These spectra of the acetylated hemoglobin adducts showed 10-11 well resolved adduct peaks, indicating that the acetylation was not highly residue specific. The chemical shift pattern observed is in general similar to that obtained recently using [1'-(13)C]aspirin as the acetylating agent (Xu, A. S. L., Macdonald, J. M., Labotka, R. J., and London, R. E. (1999) Biochim. Biophys. Acta 1432, 333-349). Blocking the 2, 3-DPG binding site with inositol hexaphosphate (IHP) resulted in a selective reduction in intensity of adduct resonances, presumably corresponding to residues located in the 2,3-DPG binding cleft. The pattern of residue protection appeared to be identical to that observed in our previous study using IHP and labeled aspirin. Pre-acetylation of hemoglobin using unlabeled MAP, followed by acetylation with [1'-(13)C]aspirin indicated a general protective effect, with the greatest reduction of intensity for resonances corresponding to acetylated residues in the 2,3-DPG binding site. These studies indicated that both MAP and aspirin exhibit similar, although not identical, acetylation profiles and target primarily the betaLys-82 residue in the 2,3-DPG binding site, as well as sites such as betaLys-59 and alphaLys-90, which are not located in the beta-cleft of hemoglobin. PMID- 10480864 TI - Two invariant tryptophans on the alpha1 subunit define domains necessary for GABA(A) receptor assembly. AB - Two invariant tryptophan residues on the N-terminal extracellular region of the rat alpha1 subunit, Trp-69 and Trp-94, are critical for the assembly of the GABA(A) (gamma-aminobutyric acid, type A) receptor into a pentamer. These tryptophans are common not only to all GABA(A) receptor subunits, but also to all ligand-gated ion channel subunits. Converting each Trp residue to Phe and Gly by site-directed mutagenesis allowed us to study the role of these invariant tryptophan residues. Mutant alpha1 subunits, coexpressed with beta2 subunits in baculovirus-infected Sf9 cells, displayed high affinity binding to [(3)H]muscimol, a GABA site ligand, but no binding to [(35)S]t-butyl bicyclophosphorothionate, a ligand for the receptor-associated ion channel. Neither [(3)H]muscimol binding to intact cells nor immunostaining of nonpermeabilized cells gave evidence of surface expression of the receptor. When expressed with beta2 and gamma2 polypeptides, the mutant alpha1 polypeptides did not form [(3)H]flunitrazepam binding sites though wild-type alpha1 polypeptides did. The distribution of the mutant receptors on sucrose gradients suggests that the effects on ligand binding result from the inability of the mutant alpha1 subunits to form pentamers. We conclude that Trp-69 and Trp-94 participate in the formation of the interface between alpha and beta subunits, but not of the GABA binding site. PMID- 10480865 TI - Characterization of a two-component alkanesulfonate monooxygenase from Escherichia coli. AB - The Escherichia coli ssuEADCB gene cluster is required for the utilization of alkanesulfonates as sulfur sources, and is expressed under conditions of sulfate or cysteine starvation. The SsuD and SsuE proteins were overexpressed and characterized. SsuE was purified to homogeneity as an N-terminal histidine-tagged fusion protein. Native SsuE was a homodimeric enzyme of M(r) 58,400, which catalyzed an NAD(P)H-dependent reduction of FMN, but it was also able to reduce FAD or riboflavin. The SsuD protein was purified to >98% purity using cation exchange, anion exchange, and hydrophobic interaction chromatography. The pure enzyme catalyzed the conversion of pentanesulfonic acid to sulfite and pentaldehyde and was able to desulfonate a wide range of sulfonated substrates including C-2 to C-10 unsubstituted linear alkanesulfonates, substituted ethanesulfonic acids and sulfonated buffers. SsuD catalysis was absolutely dependent on FMNH(2) and oxygen, and was maximal for SsuE/SsuD molar ratios of 2.1 to 4.2 in 10 mM Tris-HCl, pH 9.1. Native SsuD was a homotetrameric enzyme of M(r) 181,000. These results demonstrate that SsuD is a broad range FMNH(2) dependent monooxygenase catalyzing the oxygenolytic conversion of alkanesulfonates to sulfite and the corresponding aldehydes. SsuE is the FMN reducing enzyme providing SsuD with FMNH(2). PMID- 10480866 TI - Direct interaction of proliferating cell nuclear antigen with the p125 catalytic subunit of mammalian DNA polymerase delta. AB - The formation of a complex between DNA polymerase delta (pol delta) and its sliding clamp, proliferating cell nuclear antigen (PCNA), is responsible for the maintenance of processive DNA synthesis at the leading strand of the replication fork. In this study, the ability of the p125 catalytic subunit of DNA polymerase delta to engage in protein-protein interactions with PCNA was established by biochemical and genetic methods. p125 and PCNA were shown to co-immunoprecipitate from either calf thymus or HeLa extracts, or when they were ectopically co expressed in Cos 7 cells. Because pol delta is a multimeric protein, this interaction could be indirect. Thus, rigorous evidence was sought for a direct interaction of the p125 catalytic subunit and PCNA. To do this, the ability of recombinant p125 to interact with PCNA was established by biochemical means. p125 co-expressed with PCNA in Sf9 cells was shown to form a physical complex that can be detected on gel filtration and that can be cross-linked with the bifunctional cross-linking agent Sulfo-EGS (ethylene glycol bis (sulfosuccinimidylsuccinate)). An interaction between p125 and PCNA could also be demonstrated in the yeast two hybrid system. Overlay experiments using biotinylated PCNA showed that the free p125 subunit interacts with PCNA. The PCNA overlay blotting method was also used to demonstrate the binding of synthetic peptides corresponding to the N2 region of pol delta and provides evidence for a site on pol delta that is involved in the protein-protein interactions between PCNA and pol delta. This region contains a sequence that is a potential member of the PCNA binding motif found in other PCNA-binding proteins. These studies provide an unequivocal demonstration that the p125 subunit of pol delta interacts with PCNA. PMID- 10480867 TI - The yeast Hsp110 family member, Sse1, is an Hsp90 cochaperone. AB - In eukaryotes, production of the diverse repertoire of molecular chaperones during normal growth and in response to stress is governed by the heat shock transcription factor HSF. The HSC82 and HSP82 genes, encoding isoforms of the yeast Hsp90 molecular chaperone, were recently identified as targets of the HSF carboxyl-terminal activation domain (CTA), whose expression is required for cell cycle progression during prolonged heat stress conditions. In the present study, we have identified additional target genes of the HSF CTA, which include nearly all of the heat shock-inducible members of the Hsp90 chaperone complex, demonstrating coordinate regulation of these components by HSF. Heat shock induction of SSE1, encoding a member of the Hsp110 family of heat shock proteins, was also dependent on the HSF CTA. Disruption of SSE1 along with STI1, encoding an established subunit of the Hsp90 chaperone complex, resulted in a severe synthetic growth phenotype. Sse1 associated with partially purified Hsp90 complexes and deletion of the SSE1 gene rendered cells susceptible to the Hsp90 inhibitors macbecin and geldanamycin, suggesting functional interaction between Sse1 and Hsp90. Sse1 is required for function of the glucocorticoid receptor, a model substrate of the Hsp90 chaperone machinery, and Hsp90-based repression of HSF under nonstress conditions. Taken together, these data establish Sse1 as an integral new component of the Hsp90 chaperone complex in yeast. PMID- 10480868 TI - Regulation of a cell type-specific silencer in the human interleukin-3 gene promoter by the transcription factor YY1 and an AP2 sequence-recognizing factor. AB - Negative regulation of cytokine gene transcription is an important mechanism in maintaining homeostasis of immune function. In this study, we characterized a silencer element in the human interleukin-3 gene promoter that is responsible for the cell-specific expression of interleukin-3. This silencer activity was proposed to be mediated by an unidentified nuclear inhibitory protein (NIP). In this study, we have identified two nuclear factors that are responsible for the silencer activity in T cells. The NIP element forms four specific DNA-protein complexes (designated as complexes A-D) with the Jurkat nuclear proteins. Complex A contains a nuclear protein that shares DNA-binding specificity with the transcription factor AP2 (designated as an AP2 sequence-recognizing factor (ASRF)). Formation of this ASRF complex is required for the NIP silencer function, as mutation of the ASRF-binding site abrogated the silencer activity. Complex B contains the nuclear factor YY1 (Yin-Yang 1), whose function is to down regulate ASRF activity in the silencer. YY1 activity is supported by data from mutation and cotransfection analyses. Complexes C and D are formed by nonspecific binding proteins and do not express any regulatory activity in the NIP element. These data indicate that a cell type-specific silencer activity might be determined by a unique profile of ubiquitous transcription factors. PMID- 10480869 TI - Biochemical characterization of the interaction between the Saccharomyces cerevisiae MSH2-MSH6 complex and mispaired bases in DNA. AB - The interaction of the Saccharomyces cerevisiae MSH2-MSH6 complex with mispaired bases was analyzed using gel mobility shift assays and surface plasmon resonance methods. Under equilibrium binding conditions, MSH2-MSH6 bound to homoduplex DNA with a K(d) of 3.9 nM and bound oligonucleotide duplexes containing T:G, +1, +2, +4, and +10 insertion/deletion loop (IDL) mispairs with K(d) values of 0.20, 0.25, 11, 3.2, and 0.55 nM, respectively. Competition binding experiments using 65 different substrates revealed a 10-fold range in mispair discrimination. In general, base-base mispairs and a +1 insertion/deletion mispair were recognized better than intermediate sized insertion/deletion mispairs of 2-8 bases. Larger IDL mispairs (>8 bases) were recognized almost as well as the +1 IDL mispair. Recognition of mispairs by MSH2-MSH6 was influenced by sequence context, with the 6-nucleotide region surrounding the mispair being primarily responsible for influencing mispair recognition. Effects of sequences as far away as 15 nucleotides were also observed. Differential effects of ATP on the stability of MSH2-MSH6-mispair complexes suggested that base-base mispairs and the smaller IDL mispairs were recognized by a different binding mode than larger IDL mispairs, consistent with genetic experiments indicating that MSH2-MSH6 functions primarily in the repair of base-base and small IDL mispairs. PMID- 10480870 TI - The existence of the K(+) channel in plant mitochondria. AB - In this study, evidence is given that a number of isolated coupled plant mitochondria (from durum wheat, bread wheat, spelt, rye, barley, potato, and spinach) can take up externally added K(+) ions. This was observed by following mitochondrial swelling in isotonic KCl solutions and was confirmed by a novel method in which the membrane potential decrease due to externally added K(+) is measured fluorimetrically by using safranine. A detailed investigation of K(+) uptake by durum wheat mitochondria shows hyperbolic dependence on the ion concentration and specificity. K(+) uptake electrogenicity and the non competitive inhibition due to either ATP or NADH are also shown. In the whole, the experimental findings reported in this paper demonstrate the existence of the mitochondrial K(+)(ATP) channel in plants (PmitoK(ATP)). Interestingly, Mg(2+) and glyburide, which can inhibit mammalian K(+) channel, have no effect on PmitoK(ATP). In the presence of the superoxide anion producing system (xanthine plus xanthine oxidase), PmitoK(ATP) activation was found. Moreover, an inverse relationship was found between channel activity and mitochondrial superoxide anion formation, as measured via epinephrine photometric assay. These findings strongly suggest that mitochondrial K(+) uptake could be involved in plant defense mechanism against oxidative stress due to reactive oxygen species generation. PMID- 10480871 TI - Mechano- or acid stimulation, two interactive modes of activation of the TREK-1 potassium channel. AB - TREK-1 is a member of the novel structural class of K(+) channels with four transmembrane segments and two pore domains in tandem (1,2). TREK-1 is opened by membrane stretch and arachidonic acid. It is also an important target for volatile anesthetics (2,3). Here we show that internal acidification opens TREK 1. Indeed, lowering pH(i) shifts the pressure-activation relationship toward positive values and leads to channel opening at atmospheric pressure. The pH(i) sensitive region in the carboxyl terminus of TREK-1 is the same that is critically involved in mechano-gating as well as arachidonic acid activation. A convergence, which is dependent on the carboxyl terminus, occurs between mechanical, fatty acids and acidic stimuli. Intracellular acidosis, which occurs during brain and heart ischemia, will induce TREK-1 opening with subsequent K(+) efflux and hyperpolarization. PMID- 10480872 TI - The CLK family kinases, CLK1 and CLK2, phosphorylate and activate the tyrosine phosphatase, PTP-1B. AB - The protein-tyrosine phosphatase PTP-1B is an important regulator of intracellular protein tyrosine phosphorylation, and is itself regulated by phosphorylation. We report that PTP-1B and its yeast analog, YPTP, are phosphorylated and activated by members of the CLK family of dual specificity kinases. CLK1 and CLK2 phosphorylation of PTP-1B in vitro activated the phosphatase activity approximately 3-5-fold using either p-nitrophenol phosphate, or tyrosine-phosphorylated myelin basic protein as substrates. Co-expression of CLK1 or CLK2 with PTP-1B in HEK 293 cells led to a 2-fold stimulation of phosphatase activity in vivo. Phosphorylation of PTP-1B at Ser(50) by CLK1 or CLK2 is responsible for its enzymatic activation. These findings suggest that phosphorylation at Ser(50) by serine threonine kinases may regulate the activation of PTP-1B in vivo. We also show that CLK1 and CLK2 phosphorylate and activate the S. cerevisiae PTP-1B family member, YPTP1. CLK1 phosphorylation of YPTP1 led to a 3-fold stimulation of phosphatase activity in vitro. We demonstrate that CLK phosphorylation of Ser(83) on YPTP1 is responsible for the activation of this enzyme. These findings demonstrate that the CLK kinases activate PTP-1B family members, and this phosphatase may be an important cellular target for CLK action. PMID- 10480873 TI - Protein phosphatase 2C inactivates F-actin binding of human platelet moesin. AB - During activation of platelets by thrombin phosphorylation of Thr(558) in the C terminal domain of the membrane-F-actin linking protein moesin increases transiently, and this correlates with protrusion of filopodial structures. Calyculin A enhances phosphorylation of moesin by inhibition of phosphatases. To measure this moesin-specific activity, a nonradioactive enzyme-linked immunosorbent assay method was developed with the synthetic peptide Cys-Lys(555) Tyr-Lys-Thr(P)-Leu-Arg(560) coupled to bovine serum albumin as the substrate and moesin phosphorylation state-specific polyclonal antibodies for the detection and quantitation of dephosphorylation. Calyculin A-sensitive and -insensitive protein threonine phosphatase activities were detected in platelet lysates and separated by DEAE-cellulose chromatography. The calyculin A-sensitive enzyme was identified as a type 1 protein phosphatase. The calyculin A-insensitive enzyme activity was purified to homogeneity by phenyl- Sepharose, protamine-, and phosphonic acid peptide-agarose chromatography and characterized biochemically and immunologically as a 53-kDa protein(s) and a type 2C protein phosphatase (PP2C). Phosphorylation of Thr(558) is necessary for F-actin binding of moesin in vitro. The purified enzyme, as well as bacterially made PP2Calpha and PP2Cbeta, efficiently dephosphorylate(s) highly purified platelet phospho-moesin. This reverses the activating effect of phosphorylation, and moesin no longer co sediments with actin filaments. In vivo, regulation of these phosphatase activities are likely to influence dynamic interactions between the actin cytoskeleton and membrane constituents linked to moesin. PMID- 10480874 TI - Selective binding of steroid hormone receptors to octamer transcription factors determines transcriptional synergism at the mouse mammary tumor virus promoter. AB - Transcriptional synergism between glucocorticoid receptor (GR) and octamer transcription factors 1 and 2 (Oct-1 and Oct-2) in the induction of mouse mammary tumor virus (MMTV) transcription has been proposed to be mediated through directed recruitment of the octamer factors to their binding sites in the viral long terminal repeat. This recruitment correlates with direct binding between the GR DNA binding domain and the POU domain of the octamer factors. In present study, in vitro experiments identified several nuclear hormone receptors to have the potential to bind to the POU domains of Oct-1 and Oct-2 through their DNA binding domains, suggesting that POU domain binding may be a property shared by many nuclear hormone receptors. However, physiologically relevant binding to the POU domain appeared to be a property restricted to only a few nuclear receptors as only GR, progesterone receptor (PR), and androgen receptor (AR), were found to interact physically and functionally with Oct-1 and Oct-2 in transfected cells. Thus GR, PR, and AR efficiently promoted the recruitment of Oct-2 to adjacent octamer motifs in the cell, whereas mineralocorticoid receptor (MR), estrogen receptor alpha, and retinoid X receptor failed to facilitate octamer factor DNA binding. For MMTV, although GR and MR both induced transcription efficiently, mutation of the promoter proximal octamer motifs strongly decreased GR-induced transcription without affecting the total level of reporter gene activity in response to MR. These results suggest that the configuration of the hormone response element within the MMTV long terminal repeat may promote a dependence for the glucocorticoid response upon the recruitment of octamer transcription factors to their response elements within the viral promoter. PMID- 10480875 TI - Interaction of translation initiation factor eIF4G with eIF4A in the yeast Saccharomyces cerevisiae. AB - Eukaryotic initiation factor (eIF) 4A is an essential protein that, in conjunction with eIF4B, catalyzes the ATP-dependent melting of RNA secondary structure in the 5'-untranslated region of mRNA during translation initiation. In higher eukaryotes, eIF4A is assumed to be recruited to the mRNA through its interaction with eIF4G. However, the failure to detect this interaction in yeast brought into question the generality of this model. The work presented here demonstrates that yeast eIF4G interacts with eIF4A both in vivo and in vitro. The eIF4A-binding site was mapped to amino acids 542-883 of yeast eIF4G1. Expression in yeast cells of the eIF4G1 domain that binds eIF4A results in cell growth inhibition, and addition of this domain to an eIF4A-dependent in vitro system inhibits translation in a dose-dependent manner. Both in vitro translation and cell growth can be specifically restored by increasing the eIF4A concentration. These data demonstrate that yeast eIF4A and eIF4G interact and suggest that this interaction is required for translation and cell growth. PMID- 10480876 TI - One intact ATP-binding subunit is sufficient to support ATP hydrolysis and translocation in an ABC transporter, the histidine permease. AB - The membrane-bound complex of the Salmonella typhimurium histidine permease, a member of the ABC transporters (or traffic ATPases) superfamily, is composed of two integral membrane proteins, HisQ and HisM, and two copies of an ATP-binding subunit, HisP, which hydrolyze ATP, thus supplying the energy for translocation. The three-dimensional structure of HisP has been resolved. Extensive evidence indicates that the HisP subunits form a dimer. We investigated the mechanism of action of such a dimer, both within the complex and in soluble form, by creating heterodimers between the wild type and mutant HisP proteins. The data strongly suggest that within the complex both subunits hydrolyze ATP and that one subunit is activated by the other. In a heterodimer containing one wild type and one hydrolysis defective subunit both hydrolysis and ligand translocation occur at half the rate of the wild type. Soluble HisP also hydrolyzes ATP if one subunit is inactive; its specific activity is identical to that of the wild type, indicating that only one of the subunits in a soluble dimer is involved in hydrolysis. We show that the activating ability varies depending on the nature of the substitution of a well conserved residue, His-211. PMID- 10480877 TI - Tetrahydrobiopterin-dependent inhibition of superoxide generation from neuronal nitric oxide synthase. AB - The binding of calcium/calmodulin stimulates electron transfer between the reductase and oxygenase domains of neuronal nitric oxide synthase (nNOS). Here, we demonstrate using electron spin resonance spin-trapping with 5 diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide that pterin-free nNOS generates superoxide from the reductase and the oxygenase domain by a calcium/calmodulin dependent mechanism. Tetrahydrobiopterin (BH(4)) diminishes the formation of superoxide by a mechanism that does not cause inhibition of NADPH consumption. In contrast, BH(4) analogs 7,8-dihydrobiopterin and sepiapterin do not affect superoxide yields. L-Arginine alone inhibits the generation of superoxide by nNOS but not by C331A-nNOS mutant that has a low affinity for L-arginine. A greater decrease in superoxide yields is observed when nNOS is preincubated with L arginine. This effect is in accordance with the slow binding rates of L-arginine to NOS in the absence of BH(4). L-Arginine alone or in combination with BH(4) decreases the rates of NADPH consumption. The effect of L-arginine on superoxide yields, however, was less dramatic than that caused by BH(4) as much higher concentrations of L-arginine are necessary to attain the same inhibition. In combination, L-arginine and BH(4) inhibit the formation of superoxide generation and stimulate the formation of L-citrulline. We conclude that, in contrast to L arginine, BH(4) does not inhibit the generation of superoxide by controlling electron transfer through the enzyme but by stimulating the formation of the heme peroxo species. PMID- 10480878 TI - Stereochemical course and steady state mechanism of the reaction catalyzed by the GDP-fucose synthetase from Escherichia coli. AB - Recently the genes encoding the human and Escherichia coli GDP-mannose dehydratase and GDP-fucose synthetase (GFS) protein have been cloned and it has been shown that these two proteins alone are sufficient to convert GDP mannose to GDP fucose in vitro. GDP-fucose synthetase from E. coli is a novel dual function enzyme in that it catalyzes epimerizations and a reduction reaction at the same active site. This aspect separates fucose biosynthesis from that of other deoxy and dideoxy sugars in which the epimerase and reductase activities are present on separate enzymes encoded by separate genes. By NMR spectroscopy we have shown that GFS catalyzes the stereospecific hydride transfer of the ProS hydrogen from NADPH to carbon 4 of the mannose sugar. This is consistent with the stereospecificity observed for other members of the short chain dehydrogenase reductase family of enzymes of which GFS is a member. Additionally the enzyme is able to catalyze the epimerization reaction in the absence of NADP or NADPH. The kinetic mechanism of GFS as determined by product inhibition and fluorescence binding studies is consistent with a random mechanism. The dissociation constants determined from fluorescence studies indicate that the enzyme displays a 40-fold stronger affinity for the substrate NADPH as compared with the product NADP and utilizes NADPH preferentially as compared with NADH. This study on GFS, a unique member of the short chain dehydrogenase reductase family, coupled with that of its recently published crystal structure should aid in the development of antimicrobial or anti-inflammatory compounds that act by blocking selectin mediated cell adhesion. PMID- 10480879 TI - Villin function in the organization of the actin cytoskeleton. Correlation of in vivo effects to its biochemical activities in vitro. AB - Villin is an actin-binding protein of the intestinal brush border that bundles, nucleates, caps, and severs actin in a Ca(2+)-dependent manner in vitro. Villin induces the growth of microvilli in transfected cells, an activity that requires a carboxyl-terminally located KKEK motif. By combining cell transfection and biochemical assays, we show that the capacity of villin to induce growth of microvilli in cells correlates with its ability to bundle F-actin in vitro but not with its nucleating activity. In agreement with its importance for microfilament bundling in cells, the KKEK motif of the carboxyl-terminal F-actin binding site is crucial for bundling in vitro. In addition, substitutions of basic residues in a second site, located in the amino-terminal portion of villin, impaired its activity in cells and reduced its binding to F-actin in the absence of Ca(2+) as well as its bundling and severing activities in vitro. Altogether, these findings suggest that villin participates in the organization and stabilization of the brush border core bundle but does not initiate its assembly by nucleation of actin filaments. PMID- 10480880 TI - Muscle-specific overexpression of FAT/CD36 enhances fatty acid oxidation by contracting muscle, reduces plasma triglycerides and fatty acids, and increases plasma glucose and insulin. AB - Increasing evidence has implicated the membrane protein CD36 (FAT) in binding and transport of long chain fatty acids (FA). To determine the physiological role of CD36, we examined effects of its overexpression in muscle, a tissue that depends on FA for its energy needs and is responsible for clearing a major fraction of circulating FA. Mice with CD36 overexpression in muscle were generated using the promoter of the muscle creatine kinase gene (MCK). Transgenic (MCK-CD36) mice had a slightly lower body weight than control litter mates. This reflected a leaner body mass with less overall adipose tissue, as evidenced by magnetic resonance spectroscopy. Soleus muscles from transgenic animals exhibited a greatly enhanced ability to oxidize fatty acids in response to stimulation/contraction. This increased oxidative ability was not associated with significant alterations in histological appearance of muscle fibers. Transgenic mice had lower blood levels of triglycerides and fatty acids and a reduced triglyceride content of very low density lipoproteins. Blood cholesterol levels were slightly lower, but no significant decrease in the cholesterol content of major lipoprotein fractions was measured. Blood glucose was significantly increased, while insulin levels were similar in the fed state and higher in the fasted state. However, glucose tolerance curves, determined at 20 weeks of age, were similar in control and transgenic mice. In summary, the study documented, in vivo, the role of CD36 to facilitate cellular FA uptake. It also illustrated importance of the uptake process in muscle to overall FA metabolism and glucose utilization. PMID- 10480881 TI - Characterization of the Stat5 protease. AB - Immature myeloid cells have been shown to transduce signals through a carboxyl terminally truncated isoform of Stat5. This functionally distinct signal transducer and activator of transcription isoform is generated through a unique protein-processing event. Evaluation of numerous cell lines has determined that there is a direct correlation between the expression of truncated Stat5 and protease activity. Moreover, protease activity is found only in the myeloid and not in lymphoid progenitors. To further characterize the protease small quantities have been purified to near homogeneity. Studies on this purified material indicate that the protease has an apparent molecular mass of approximately 25 kDa and is active over a wide range of pH values. The protease will also cleave both activated (i.e. tyrosine-phosphorylated) and inactivate Stat5. Although this activity is sensitive to phenylmethylsulfonyl fluoride, it is notably not sensitive to several other serine protease inhibitors. Additional studies have led to the identification of the unique site where the protease cleaves Stat5. Mutagenesis of this site renders Stat5 resistant to cleavage. Consistent with the model that Stat5 cleavage is important for early myeloid development, introduction of a "non-cleavable" isoform of Stat5 into FDC-P1 cells (a myeloid progenitor line) leads to significant phenotypic changes. PMID- 10480882 TI - p70 S6 kinase-mediated protein synthesis is a critical step for vascular endothelial cell proliferation. AB - In this work, we analyzed the role of the PI3K-p70 S6 kinase (S6K) signaling cascade in the stimulation of endothelial cell proliferation. We found that inhibitors of the p42/p44 MAPK pathway (PD98059) and the PI3K-p70 S6K pathway (wortmannin, Ly294002, and rapamycin) all block thymidine incorporation stimulated by fetal calf serum in the resting mouse endothelial cell line 1G11. The action of rapamycin can be generalized, since it completely inhibits the mitogenic effect of fetal calf serum in primary endothelial cell cultures (human umbilical vein endothelial cells) and another established capillary endothelial cell line (LIBE cells). The inhibitory effect of rapamycin is only observed when the inhibitor is added at the early stages of G(0)-G(1) progression, suggesting an inhibitory action early in G(1). Rapamycin completely inhibits growth factor stimulation of protein synthesis, which perfectly correlates with the inhibition of cell proliferation. In accordance with its inhibitory action on protein synthesis, activation of cyclin D1 and p21 proteins by growth factors is also blocked by preincubation with rapamycin. Expression of a p70 S6K mutant partially resistant to rapamycin reverses the inhibitory effect of the drug on DNA synthesis, indicating that rapamycin action is via p70 S6K. Thus, in vascular endothelial cells, activation of protein synthesis via p70 S6K is an essential step for cell cycle progression in response to growth factors. PMID- 10480883 TI - The targeted disruption of both alleles of RARbeta(2) in F9 cells results in the loss of retinoic acid-associated growth arrest. AB - F9 teratocarcinoma cell lines, carrying one or two disrupted alleles of the RARbeta(2) gene, were generated by homologous recombination to study the role of RARbeta(2) in mediating the effects of retinoids on cell growth and differentiation. Retinoic acid (RA) does not induce growth arrest of the RARbeta(2)-/- cells, whereas the F9 WT and RARbeta(2)+/- heterozygote lines undergo RA-induced growth arrest. The RARbeta(2)+/- lines also exhibit a faster cell cycle transit time in the absence of RA. The RARbeta(2)-/- stem cells exhibit an altered morphology when compared with the F9 WT parent line, and after RA treatment, the RARbeta(2)-/- cells do not exhibit a fully differentiated cell morphology. As compared with F9 WT cells, the RARbeta-/- cells exhibited a markedly lower induction of several early RA-responsive genes and no induction of laminin B1, a late response gene. The induction of RA metabolism in the F9 RARbeta(2)-/- cells following differentiation was not impaired. The research presented here, and prior research suggest that RARbeta is required for RA induced growth arrest in a variety of cell types and that RARbeta also functions in mediating late responses to RA. These findings are significant in view of the reduced expression of RARbeta transcripts in a number of different types of human carcinomas. PMID- 10480884 TI - C172S substitution in the chloroplast-encoded large subunit affects stability and stress-induced turnover of ribulose-1,5-bisphosphate carboxylase/oxygenase. AB - Previous work has indicated that the turnover of chloroplast ribulose-1,5 bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1. 39) may be controlled by the redox state of certain cysteine residues. To test this hypothesis, directed mutagenesis and chloroplast transformation were employed to create a C172S substitution in the Rubisco large subunit of the green alga Chlamydomonas reinhardtii. The C172S mutant strain was not substantially different from the wild type with respect to growth rate, and the purified mutant enzyme had a normal circular dichroism spectrum. However, the mutant enzyme was inactivated faster than the wild-type enzyme at 40 and 50 degrees C. In contrast, C172S mutant Rubisco was more resistant to sodium arsenite, which reacts with vicinal dithiols. The effect of arsenite may be directed to the cysteine 172/192 pair that is present in the wild-type enzyme, but absent in the mutant enzyme. The mutant enzyme was also more resistant to proteinase K in vitro at low redox potential. Furthermore, oxidative (hydrogen peroxide) or osmotic (mannitol) stress-induced degradation of Rubisco in vivo was delayed in C172S mutant cells relative to wild-type cells. Thus, cysteine residues could play a role in regulating the degradation of Rubisco under in vivo stress conditions. PMID- 10480885 TI - The sulfonium ion linkage in myeloperoxidase. Direct spectroscopic detection by isotopic labeling and effect of mutation. AB - The heme group of myeloperoxidase is covalently linked via two ester bonds to the protein and a unique sulfonium ion linkage involving Met(243). Mutation of Met(243) into Thr, Gln, and Val, which are the corresponding residues of eosinophil peroxidase, lactoperoxidase, and thyroid peroxidase, respectively, and into Cys was performed. The Soret band in the optical absorbance spectrum in the oxidized mutants is now found at approximately 411 nm. Both the pyridine hemochrome spectra and resonance Raman spectra of the mutants are affected by the mutation. In the Met(243) mutants the affinity for chloride has decreased 100 fold. All mutants have lost their chlorination activity, except for the M243T mutant, which still has 15% activity left. By Fourier transform infared difference spectroscopy it was possible to specifically detect the (13)CD(3) labeled methionyl sulfonium ion linkage. We conclude that the sulfonium ion linkage serves two roles. First, it serves as an electron-withdrawing substituent via its positive charge, and, second, together with its neighboring residue Glu(242), it appears to be responsible for the lower symmetry of the heme group and distortion from the planar conformation normally seen in heme-containing proteins. PMID- 10480886 TI - Shear stress stimulation of p130(cas) tyrosine phosphorylation requires calcium dependent c-Src activation. AB - Fluid shear stress (flow) modulates endothelial cell function via specific intracellular signaling events. Previously we showed that flow activated ERK1/2 in an integrin-dependent manner (Takahashi, M., and Berk, B. C. (1996) J. Clin. Invest. 98, 2623-2631). p130 Crk-associated substrate (Cas), a putative c-Src substrate, was originally identified as a highly phosphorylated protein that is localized to focal adhesions and acts as an adapter protein. Recent reports have shown that Cas is important in cardiovascular development and actin filament assembly. Flow (shear stress = 12 dynes/cm(2)) stimulated Cas tyrosine phosphorylation within 1 min in human umbilical vein endothelial cells. Phosphorylation peaked at 5 min (3.5 +/- 0.7-fold) and was sustained to 20 min. Tyrosine phosphorylation of Cas was functionally important because flow stimulated association of Cas with Crk in a time- and force-dependent manner. Flow-mediated activation of c-Src, phosphorylation of Cas, and association of Cas with Crk were all inhibited by calcium chelation and pretreatment with the Src family-specific tyrosine kinase inhibitor PP1. To determine the role of c-Src in flow-stimulated phosphorylation of Cas, we transduced cells with adenovirus encoding kinase-inactive Src. Expression of kinase-inactive Src prevented flow induced Cas tyrosine phosphorylation but not ERK1/2 activation. Calcium-dependent activation of c-Src and tyrosine phosphorylation of Cas defines a new flow stimulated signal pathway, different from ERK1/2 activation. This pathway may be involved in focal adhesion remodeling and actin filament assembly. PMID- 10480887 TI - Glycosylphosphatidylinositol-anchored proteins play an important role in the biogenesis of the Alzheimer's amyloid beta-protein. AB - The Alzheimer's amyloid protein (Abeta) is released from the larger amyloid beta protein precursor (APP) by unidentified enzymes referred to as beta- and gamma secretase. beta-Secretase cleaves APP on the amino side of Abeta producing a large secreted derivative (sAPPbeta) and an Abeta-bearing C-terminal derivative that is subsequently cleaved by gamma-secretase to release Abeta. Alternative cleavage of the APP by alpha-secretase at Abeta16/17 releases the secreted derivative sAPPalpha. In yeast, alpha-secretase activity has been attributed to glycosylphosphatidylinositol (GPI)-anchored aspartyl proteases. To examine the role of GPI-anchored proteins, we specifically removed these proteins from the surface of mammalian cells using phosphatidylinositol-specific phospholipase C (PI-PLC). PI-PLC treatment of fetal guinea pig brain cultures substantially reduced the amount of Abeta40 and Abeta42 in the medium but had no effect on sAPPalpha. A mutant CHO cell line (gpi85), which lacks GPI-anchored proteins, secreted lower levels of Abeta40, Abeta42, and sAPPbeta than its parental line (GPI+). When this parental line was treated with PI-PLC, Abeta40, Abeta42, and sAPPbeta decreased to levels similar to those observed in the mutant line, and the mutant line was resistant to these effects of PI-PLC. These findings provide strong evidence that one or more GPI-anchored proteins play an important role in beta-secretase activity and Abeta secretion in mammalian cells. The cell-surface GPI-anchored protein(s) involved in Abeta biogenesis may be excellent therapeutic target(s) in Alzheimer's disease. PMID- 10480888 TI - A rho exchange factor mediates thrombin and Galpha(12)-induced cytoskeletal responses. AB - Thrombin induces astrocytoma cell rounding through a Rho-dependent pathway (Majumdar, M., Seasholtz, T. M., Goldstein, D., de Lanerolle, P., and Brown, J. H. (1998) J. Biol. Chem. 273, 10099-10106). The involvement of the G(12) family of G proteins and the role of specific Rho exchange factors in transducing signals from the thrombin receptor to Rho-dependent cytoskeletal responses was examined. Microinjection of cDNAs for activated Galpha(12) or Galpha(13) induced cell rounding, and antibodies to Galpha(12) or Galpha(13) blocked the response to thrombin. In contrast, activation or inhibition of Galpha(q) function had relatively little effect. The cytoskeletal response to Galpha(12) was inhibited by microinjection of C3 exoenzyme, indicating Rho dependence. Two Rho-specific guanine nucleotide exchange factors (GEFs), oncogenic lbc and p115, increased the percentage of rounded cells 4-5-fold, and this was inhibited by C3. Mutant GEFs lacking the Dbl homology (DH) domain required for exchange factor activity failed to induce cell rounding. However, the DH mutants of lbc and p115 were efficacious inhibitors of rounding induced by thrombin or Galpha(12). The effects of lbc were dependent on an intact pleckstrin homology domain, which may be required for appropriate targeting of the Rho-GEF. These findings identify the Galpha(12) protein family as transducers of thrombin signaling to the cytoskeleton and provide the first evidence that a Rho-GEF transduces signals between G protein coupled receptors and Rho-mediated cytoskeletal responses. PMID- 10480889 TI - Transcription dependence and the roles of two excision repair pathways for UV damage in fission yeast Schizosaccharomyces pombe. AB - Fission yeasts Schizosaccharomyces pombe possess two types of excision repair systems for UV-induced DNA damage, nucleotide excision repair (NER) and UV damaged DNA endonuclease (UVDE)-dependent excision repair (UVER). Despite its high efficiency in damage removal, UVER defects have less effect on UV survival than NER defects. To understand the differential roles of two pathways, we examined strand-specific damage removal at the myo2 and rpb2 loci. Although NER removes cyclobutane pyrimidine dimers from the transcribed strand more rapidly than from the nontranscribed strand, UVER repairs cyclobutane pyrimidine dimers equally on both strands and at a much higher rate than NER. The low rate of damage removal from the nontranscribed strand in the absence of UVER indicates inefficient global genome repair (GGR) in this organism and a possible function of UVER as an alternative to GGR. Disruption of rhp26, the S. pombe homolog of CSB/RAD26, eliminated the strand bias of NER almost completely and resulted in a significant increase of UV sensitivity of cells in a uvdeDelta background. We suggest that the combination of transcription-coupled repair of NER and rapid UVER contributes to UV survival in growing S. pombe cells, which is accomplished by transcription-coupled repair and GGR in other organisms. PMID- 10480890 TI - Direct kinetic evidence for folding via a highly compact, misfolded state. AB - The 2 S seed storage protein, sunflower albumin 8 (SFA-8), contains an unusually high proportion of hydrophobic residues including 16 methionines (some of which may form a surface hydrophobic patch) in a disulfide cross-linked, alpha-helical structure. Circular dichroism and fluorescence spectroscopy show that SFA-8 is highly stable to denaturation by heating or chaotropic agents, the latter resulting in a reversible two-state unfolding transition. The small m(U) (-4.7 M( 1) at 10 degrees C) and DeltaC(p) (-0.95 kcal mol(-1) K(-1)) values indicate that relatively little nonpolar surface of the protein is exposed during unfolding. Commensurate with the unusual distribution of hydrophobic residues, stopped-flow fluorescence data show that the folding pathway of SFA-8 is highly atypical, in that the initial product of the rapid collapse phase of folding is a compact nonnative state (or collection of nonnative states) that must unfold before acquiring the native conformation. The inhibited folding reaction of SFA-8, in which the misfolded state (m(M) = -0.95 M(-1) at 10 degrees C) is more compact than the transition state for folding (m(T) = -2.5 M(-1) at 10 degrees C), provides direct kinetic evidence for the transient misfolding of a protein. PMID- 10480891 TI - Salt links dominate affinity of antibody HyHEL-5 for lysozyme through enthalpic contributions. AB - The binding of murine monoclonal antibody HyHEL-5 to lysozyme has been the subject of extensive crystallographic, computational, and experimental investigations. The complex of HyHEL-5 with hen egg lysozyme (HEL) features salt bridges between Fab heavy chain residue Glu(50), and Arg(45) and Arg(68) of HEL. This interaction has been predicted to play a dominant role in the association on the basis of molecular electrostatics calculations. The association of aspartic acid and glutamine mutants at position 50(H) of the cloned HyHEL-5 Fab with HEL and bobwhite quail lysozyme (BQL), an avian variant bearing an Arg(68) --> Lys substitution in the epitope, was characterized by isothermal titration calorimetry and sedimentation equilibrium. Affinities for HEL were reduced by 400 fold (E50(H)D) and 40,000-fold (E50(H)Q) (DeltaDeltaG degrees estimated at 4.0 and 6.4 kcal mol(-1), respectively). The same mutations reduce affinity for BQL by only 7- and 55-fold, respectively, indicating a reduced specificity for HEL. The loss of affinity upon mutation is in each case primarily due to an unfavorable change in the enthalpy of the interaction; the entropic contribution is virtually unchanged. An enthalpy-entropy compensation exists for each interaction; DeltaH degrees decreases, while DeltaS degrees increases with temperature. The DeltaCp for each mutant interaction is less negative than the wild-type. Mutant-cycle analysis suggests the mutations present in the HyHEL-5 Fab mutants are linked to those present in the BQL with coupling energies between 3 and 4 kcal mol(-1). PMID- 10480892 TI - Isolation and characterization of bone morphogenetic protein-binding proteins from the early Xenopus embryo. AB - Using a surface plasmon resonance biosensor as a sensitive and specific monitor, we have isolated two distinct bone morphogenetic protein (BMP)-binding proteins, and identified them as lipovitellin 1 and Ep45, respectively. Lipovitellin 1 is an egg yolk protein that is processed from vitellogenin. Both vitellogenin and Ep45 are synthesized under estrogen control in the liver, secreted, and taken up by developing oocytes. In this paper, we have shown that of the TGF-beta family members tested, Ep45 can bind only to BMP-4, whereas lipovitellin 1 can bind to both BMP-4 and activin A. Because of this difference in specificity, we have focused on and further studied Ep45. Kinetic parameters were determined by surface plasmon resonance studies and showed that Ep45 associated rapidly with BMP-4 (k(a) = 1.06 x 10(4) M(-1)s(-1)) and dissociated slowly (k(d) = 1.6 x 10( 4) s(-1)). In Xenopus embryos microinjected with Ep45 mRNA, Ep45 blocked the ability of follistatin to inhibit BMP activity and to induce a secondary body axis in a dose-dependent manner, whereas it had no effect on other BMP antagonists, chordin and noggin. These results support the possibility that Ep45 interacts with BMP to modulate its activities in vivo. PMID- 10480893 TI - Requirement of an E1A-sensitive coactivator for long-range transactivation by the beta-globin locus control region. AB - Four erythroid-specific DNase I-hypersensitive sites at the 5'-end of the beta globin locus confer high-level transcription to the beta-globin genes. To identify coactivators that mediate long-range transactivation by this locus control region (LCR), we assessed the influence of E1A, an inhibitor of the CBP/p300 histone acetylase, on LCR function. E1A strongly inhibited transactivation of Agamma- and beta-globin promoters by the HS2, HS2-HS3, and HS1 HS4 subregions of the LCR in human K562 and mouse erythroleukemia cells. Short- and long-range transactivation mediated by the LCR were equally sensitive to E1A. The E1A sensitivity was apparent in transient and stable transfection assays, and E1A inhibited expression of the endogenous gamma-globin genes. Only sites for NF E2 within HS2 were required for E1A sensitivity in K562 cells, and E1A abolished transactivation mediated by the activation domain of NF-E2. E1A mutants defective in CBP/p300 binding only weakly inhibited HS2-mediated transactivation, whereas a mutant defective in retinoblastoma protein binding strongly inhibited transactivation. Expression of CBP/p300 potentiated HS2-mediated transactivation. Moreover, expression of GAL4-CBP strongly increased transactivation of a reporter containing HS2 with a GAL4 site substituted for the NF-E2 sites. Thus, we propose that a CBP/p300-containing coactivator complex is the E1A-sensitive factor important for LCR function. PMID- 10480895 TI - The actin cytoskeleton regulates LFA-1 ligand binding through avidity rather than affinity changes. AB - To elucidate the role of the cytoskeleton regulating avidity or affinity changes in the leukocyte adhesion receptor lymphocyte function-associated antigen-1 (LFA 1) (alpha(L)beta(2)), we generated mutant cytoplasmic LFA-1 receptors and expressed these into the erythroleukemic cell line K562. We determined whether intercellular adhesion molecule-1 (ICAM-1)-mediated adhesion of LFA-1, lacking parts of its cytoplasmic tails, is regulated through receptor diffusion/clustering and/or by altered ligand binding affinity. All cytoplasmic deletion mutants that lack the complete beta(2) cytoplasmic tail and/or the conserved KVGFFKR sequence in the alpha(L) cytoplasmic tail were constitutively active and expressed high levels of the activation epitopes NKI-L16 and M24. Surprisingly, whereas these mutants showed a clustered cell surface distribution of LFA-1, the ligand-binding affinity as measured by titration of soluble ligand ICAM-1 remained unaltered. The notion that redistribution of LFA-1 does not alter ligand-binding affinity is further supported by the finding that disruption of the cytoskeleton by cytochalasin D did not alter the binding affinity nor adhesion to ICAM-1 of these mutants. Most cytoplasmic deletion mutants that spontaneously bound ICAM-1 were not capable to spread on ICAM-1, demonstrating that on these mutants LFA-1 is not coupled to the actin cytoskeleton. From these data we conclude that LFA-1-mediated cell adhesion to ICAM-1 is predominantly regulated by receptor clustering and that affinity alterations do not necessarily coincide with strong ICAM-1 binding. PMID- 10480894 TI - RGS1 is expressed in monocytes and acts as a GTPase-activating protein for G protein-coupled chemoattractant receptors. AB - The leukocyte response to chemoattractants is transduced by the interaction of transmembrane receptors with GTP-binding regulatory proteins (G-proteins). RGS1 is a member of a protein family constituting a newly appreciated and large group of proteins that act as deactivators of G-protein signaling pathways by accelerating the GTPase activity of G-protein alpha subunits. We demonstrate here that RGS1 is expressed in human monocytes; by immunofluorescence and subcellular fractionation RGS1 was localized to the plasma membrane. By using a mixture of RGS1 and plasma membranes, we were able to demonstrate GAP activity of RGS1 on receptor-activated G-proteins; RGS1 did not affect ligand-stimulated GDP-GTP exchange. We found that RGS1 desensitizes a variety of chemotactic receptors including receptors for N-formyl-methionyl-leucyl-phenylalanine, leukotriene B4, and C5a. Interaction of RGS proteins and ligand-induced G-protein signaling can be demonstrated by determining GTPase activity using purified RGS proteins and plasma membranes. PMID- 10480896 TI - Characterization and chromosomal localization of USP3, a novel human ubiquitin specific protease. AB - Conjugation to the small eukaryotic protein ubiquitin can functionally modify or target proteins for degradation by the proteasome. Removal of the ubiquitin modification, or deubiquitination, is performed by ubiquitin-specific proteases and is an important mechanism regulating this pathway. Here we describe a novel human ubiquitin-specific protease, USP3, initially identified as a partial cDNA clone similar to one of two highly conserved sequence regions common to all ubiquitin-specific proteases. We have isolated a complete USP3 cDNA clone containing both of these conserved sequence regions. The USP3 gene appears to be single copy and maps to human chromosome 15q22.3. A USP3 probe detects two mRNA transcripts, one of which corresponds in length to the cDNA. Both are expressed at low levels in all tissues examined, with highest expression in pancreas. The USP3 protein is a functional ubiquitin-specific protease in vitro, and is able to inhibit ubiquitin-dependent degradation of both an N-end Rule substrate and abnormal endogenous proteins in yeast. USP3 is also only the second known ubiquitin-specific protease capable of efficiently cleaving a ubiquitin-proline bond. PMID- 10480897 TI - A novel family of yeast chaperons involved in the distribution of V-ATPase and other membrane proteins. AB - Null mutations in genes encoding V-ATPase subunits in Saccharomyces cerevisiae result in a phenotype that is unable to grow at high pH and is sensitive to high and low metal-ion concentrations. Treatment of these null mutants with ethylmethanesulfonate causes mutations that suppress the V-ATPase null phenotype, and the mutant cells are able to grow at pH 7.5. The suppressor mutants were denoted as svf (suppressor of V-ATPase function). The frequency of svf is relatively high, suggesting a large target containing several genes for the ethylmethanesulfonate mutagenesis. The suppressors' frequency is dependent on the individual genes that were inactivated to manifest the V-ATPase null mutation. The svf mutations are recessive, because crossing the svf mutants with their corresponding V-ATPase null mutants resulted in diploid strains that are unable to grow at pH 7.5. A novel gene family in which null mutations cause pleiotropic effects on metal-ion resistance or sensitivity and distribution of membrane proteins in different targets was discovered. The family was defined as VTC (Vacuolar Transporter Chaperon) and it contains four genes in the S. cerevisiae genome. Inactivation of one of them, VTC1, in the background of V-ATPase null mutations resulted in svf phenotype manifested by growth at pH 7.5. Deletion of the VTC1 gene (DeltaVTC1) results in a reduced amount of V-ATPase in the vacuolar membrane. These mutant cells fail to accumulate quinacrine into their vacuoles, but they are able to grow at pH 7.5. The VTC1 null mutant also results in a reduced amount of the plasma membrane H(+)-ATPase (Pma1p) in membrane preparations and possibly mis-targeting. This observation may provide an explanation for the svf phenotype in the double disruptant mutants of DeltaVTC1 and DeltaVMA subunits. PMID- 10480898 TI - Pax3 and regulation of the melanocyte-specific tyrosinase-related protein-1 promoter. AB - Previous work has established that the melanocyte-specific tyrosinase-related protein-1 (TRP-1) promoter is regulated positively by the microphthalmia associated transcription factor Mitf, acting through the conserved M box and negatively by the T-box factor Tbx2, which can bind two "melanocyte-specific elements" termed the MSEu and MSEi. Both the MSEu and MSEi, which share a 6-base pair GTGTGA consensus, are also recognized by a previously unidentified melanocyte-specific factor, MSF. Here we show using a combination of DNA binding assays, proteolytic clipping, and anti-Pax3 antibodies that MSF is indistinguishable from Pax3, a paired homeodomain transcription factor implicated genetically in melanocyte development and the regulation of the Mitf promoter. Consistent with Pax3 being able to bind the TRP-1 promoter, Pax3 is expressed in melanocytes and melanomas, and TRP-1 promoter activity is up-regulated by Pax3. The results identify a novel role for Pax3 in the expression of TRP-1, and the potential role of Pax3 in the melanocyte lineage is discussed. PMID- 10480899 TI - Suppression of ultraviolet irradiation-induced apoptosis by overexpression of focal adhesion kinase in Madin-Darby canine kidney cells. AB - Focal adhesion kinase (FAK) has been implicated to play a role in suppression of apoptosis. In this study, we have demonstrated that UV irradiation induced cleavage of FAK and two of its interacting proteins Src and p130(Cas) in Madin Darby canine kidney cells, concomitant with an increase in cell death. The cleavage of these proteins upon UV irradiation was completely inhibited by ZVAD FMK, a broad range inhibitor of caspases, and apparently delayed by Bcl2 overexpression. To examine if FAK plays a role in suppressing UV-induced apoptosis, stable Madin-Darby canine kidney cell lines overexpressing FAK were established. Our results showed that a marked (30-40%) increase in cell survival upon UV irradiation was achieved by this strategy. In our efforts to determine the mechanism by which FAK transduces survival signals to the downstream, we found that a FAK mutant deficient in binding to phosphatidylinositol 3-kinase failed to promote cell survival. Moreover, the expression of the Src homology 3 domain of p130(Cas), which competed with endogenous p130(Cas) for FAK binding, abrogated the FAK-promoted cell survival. Together, these results suggest that the integrity of FAK and its binding to phosphatidylinositol 3-kinase and p130(Cas) are required for FAK to exert its antiapoptotic function. PMID- 10480900 TI - Tryptophan 409 controls the activity of neuronal nitric-oxide synthase by regulating nitric oxide feedback inhibition. AB - The heme of neuronal nitric-oxide synthase participates in oxygen activation but also binds self-generated NO during catalysis resulting in reversible feedback inhibition. We utilized point mutagenesis to investigate if a conserved tryptophan residue (Trp-409), which engages in pi-stacking with the heme and hydrogen bonds to its axial cysteine ligand, helps control catalysis and regulation by NO. Surprisingly, mutants W409F and W409Y were hyperactive compared with the wild type regarding NO synthesis without affecting cytochrome c reduction, reductase-independent N-hydroxyarginine oxidation, or Arg and tetrahydrobiopterin binding. In the absence of Arg, NADPH oxidation measurements showed that electron flux through the heme was actually slower in the Trp-409 mutants than in wild-type nNOS. However, little or no NO complex accumulated during NO synthesis by the mutants, as opposed to the wild type. This difference was potentially related to mutants forming unstable 6-coordinate ferrous-NO complexes under anaerobic conditions even in the presence of Arg and tetrahydrobiopterin. Thus, Trp-409 mutations minimize NO feedback inhibition by preventing buildup of an inactive ferrous-NO complex during the steady state. This overcomes the negative effect of the mutation on electron flux and results in hyperactivity. Conservation of Trp-409 among different NOS suggests that the ability of this residue to regulate heme reduction and NO complex formation is important for enzyme physiologic function. PMID- 10480901 TI - Isoprenylcysteine-O-carboxyl methyltransferase regulates aldosterone-sensitive Na(+) reabsorption. AB - The Xenopus laevis distal tubule epithelial cell line A6 was used as a model epithelia to study the role of isoprenylcysteine-O-carboxyl methyltransferase (pcMTase) in aldosterone-mediated stimulation of Na(+) transport. Polyclonal antibodies raised against X. laevis pcMTase were immunoreactive with a 33-kDa protein in whole cell lysate. These antibodies were also reactive with a 33-kDa product from in vitro translation of the pcMTase cDNA. Aldosterone application increased pcMTase activity resulting in elevation of total protein methyl esterification in vivo, but pcMTase protein levels were not affected by steroid, suggesting that aldosterone increased activity independent of enzyme number. Inhibition of pcMTase resulted in a reduction of aldosterone-induced Na(+) transport demonstrating the necessity of pcMTase-mediated transmethylation for steroid induced Na(+) reabsorption. Transfection with an eukaryotic expression construct containing pcMTase cDNA increased pcMTase protein level and activity. This resulted in potentiation of the natriferic actions of aldosterone. However, overexpression did not change Na(+) reabsorption in the absence of steroid, suggesting that pcMTase activity is not limiting Na(+) transport in the absence of steroid, but that subsequent to aldosterone addition, pcMTase activity becomes limiting. These results suggest that a critical transmethylation is necessary for aldosterone-induction of Na(+) transport. It is likely that the protein catalyzing this methylation is isoprenylcysteine-O-carboxyl methyltransferase and that aldosterone activates pcMTase without affecting transferase expression. PMID- 10480902 TI - Augmentation of ultraviolet B radiation-induced tumor necrosis factor production by the epidermal platelet-activating factor receptor. AB - Ultraviolet B radiation (UVB) has been shown to damage human keratinocytes in part by inducing oxidative stress and cytokine production. Indeed, UVB-induced production of the pro-inflammatory and cytotoxic cytokine tumor necrosis factor alpha (TNF-alpha) has been implicated in the epidermal damage seen in response to acute solar radiation. Though the lipid mediator platelet-activating factor (PAF) is synthesized in response to oxidative stress, and keratinocytes express PAF receptors linked to cytokine biosynthesis, it is not known whether PAF is involved in UVB-induced epidermal cell cytokine production. These studies examined the role of the PAF system in UVB-induced epidermal cell TNF-alpha biosynthesis using a novel model system created by retroviral-mediated transduction of the PAF receptor-negative human epidermal cell line KB with the human PAF receptor (PAF-R). Treatment of PAF-R-expressing KB cells with the metabolically stable PAF-R agonist carbamoyl-PAF resulted in increased TNF-alpha mRNA and protein, indicating that activation of the epidermal PAF-R was linked to TNF-alpha production. UVB irradiation of PAF-R-expressing KB cells resulted in significant increases in both TNF-alpha mRNA and protein in comparison to UVB treated control KB cells. However, UVB treatment up-regulated cyclooxygenase-2 mRNA levels to the same extent in both PAF-R-expressing and control KB cells. Pretreatment with the antioxidant vitamin E or the PAF-R antagonists WEB 2086 and A-85783 inhibited UVB-induced TNF-alpha production in the PAF-R-positive but not control KB cells. These studies suggest that the epidermal PAF-R may be a pharmacological target for UVB in skin. PMID- 10480903 TI - Characterization of FIM-FGFR1, the fusion product of the myeloproliferative disorder-associated t(8;13) translocation. AB - The t(8;13) translocation found in a rare type of stem cell myeloproliferative disorder generates a constitutively activated tyrosine kinase containing N terminal sequence encoded by the FIM gene linked to the FGFR1 kinase domain. Here we have further characterized FIM and FIM-FGFR1 proteins. Firstly, we have studied their respective subcellular localization. We show that FIM has nuclear and nucleolar localization, whereas FIM-FGFR1 is mainly cytoplasmic. Within the nucleolus, FIM colocalizes with the upstream binding factor in interphasic cells, indicating that FIM may be involved in the regulation of rRNA transcription. We demonstrate that the targetting of FIM to the nucleus depends upon its C-terminal region, which is absent in the cytoplasmic FIM-FGFR1 protein. Secondly, we demonstrate that FIM-FGFR1 has constitutive dimerization capability mediated by the FIM N-terminal sequences. Finally, we show that FIM-FGFR1 promotes survival of pro-B Ba/F3 cells after interleukin-3 withdrawal, whereas ligand-activated FGFR1 induced not only cell survival but also interleukin-3 independence. Taken together, these results indicate that FIM-FGFR1 is activated by dimerization as a cytoplasmic kinase and suggest that FIM-FGFR1 partially signals through the FGFR1 pathways. PMID- 10480904 TI - A candidate target for G protein action in brain. AB - An effector candidate for G protein action, GRIN1, was identified by screening a cDNA expression library with phosphorylated GTPgammaS-G(z)alpha as a probe. GRIN1 is a novel protein without substantial homology to known protein domains. It is expressed largely in brain and binds specifically to activated G(z)alpha, G(o)alpha, and G(i)alpha through its carboxyl-terminal region. The protein KIAA0514 (GRIN2) is homologous to GRIN1 at its carboxyl terminus and also binds to activated G(o)alpha. Both GRIN1 and G(o)alpha are membrane-bound proteins that are enriched in the growth cones of neurites. Coexpression of GRIN1 or GRIN2 with activated G(o)alpha causes formation of a network of fine processes in Neuro2a cells, suggesting that these pathways may function downstream of G(o)alpha to control growth of neurites. PMID- 10480905 TI - Ace is a collagen-binding MSCRAMM from Enterococcus faecalis. AB - A putative collagen-binding MSCRAMM, Ace, of Enterococcus faecalis was identified by searching bacterial genome data bases for proteins containing domains homologous to the ligand-binding region of Cna, the collagen-binding MSCRAMM from Staphylococcus aureus. Ace was predicted to have a molecular mass of 71 kDa and contains features characteristic of cell surface proteins on Gram-positive bacteria, including a LPXTG motif for cross-linking to the cell wall. The N terminal region of Ace contained a region (residues 174-319) in which 56% of the residues are identical or similar when compared with the minimal ligand-binding region of Cna (Cna 151-318); the remainder of the Ace A domain has 46% similarity with the corresponding region of the Cna A domain. Antibodies raised against recombinant Ace A domain were used to verify the cell surface expression of Ace on E. faecalis. These antibodies also effectively inhibited the adhesion of enterococcal cells to a collagen substrate, suggesting that Ace is a functional collagen-binding MSCRAMM. Structural modeling of the conserved region in Ace (residues 174-319) suggested a structure very similar to that reported for residues 151-318 of the Cna collagen-binding domain in which the ligand-binding site was identified as a trench transversing a beta-sheet face (Symersky, J., Patti, J. M., Carson, M., House-Pompeo, K., Teale, M., Moore, D., Jin, L., DeLucas, L. J., Hook, M., and Narayana, S. V. L. (1997) Nat. Struct. Biol. 10, 833-838). Biochemical analyses of recombinant Ace and Cna A domains supported the modeling data in that the secondary structures were similar as determined by CD spectroscopy and both proteins bound at multiple sites in type I collagen with micromolar affinities, but with different apparent kinetics. We conclude that Ace is a collagen-binding MSCRAMM on enterococci and is structurally and functionally related to the staphylococcal Cna protein. PMID- 10480907 TI - Crucial role of Lys(423) in the electron transfer of neuronal nitric-oxide synthase. AB - Nitric-oxide synthase (NOS) is composed of an oxygenase domain having cytochrome P450-type heme active site and a reductase domain having FAD- and FMN-binding sites. To investigate the route of electron transfer from the reductase domain to the heme, we generated mutants at Lys(423) in the heme proximal site of neuronal NOS and examined the catalytic activities, electron transfer rates, and NADPH oxidation rates. A K423E mutant showed no NO formation activity (<0.1 nmol/min/nmol heme), in contrast with that (72 nmol/min/nmol heme) of the wild type enzyme. The electron transfer rate (0.01 min(-1)) of the K423E on addition of excess NADPH was much slower than that (>10 min(-1)) of the wild type enzyme. From the crystal structure of the oxygenase domain of endothelial NOS, Lys(423) of neuronal NOS is likely to interact with Trp(409) which lies in contact with the heme plane and with Cys(415), the axial ligand. It is also exposed to solvent and lies in the region where the heme is closest to the protein surface. Thus, it seems likely that ionic interactions between Lys(423) and the reductase domain may help to form a flavin to heme electron transfer pathway. PMID- 10480906 TI - Mice lacking the basolateral Na-K-2Cl cotransporter have impaired epithelial chloride secretion and are profoundly deaf. AB - In chloride-secretory epithelia, the basolateral Na-K-2Cl cotransporter (NKCC1) is thought to play a major role in transepithelial Cl(-) and fluid transport. Similarly, in marginal cells of the inner ear, NKCC1 has been proposed as a component of the entry pathway for K(+) that is secreted into the endolymph, thus playing a critical role in hearing. To test these hypotheses, we generated and analyzed an NKCC1-deficient mouse. Homozygous mutant (Nkcc1(-/-)) mice exhibited growth retardation, a 28% incidence of death around the time of weaning, and mild difficulties in maintaining their balance. Mean arterial blood pressure was significantly reduced in both heterozygous and homozygous mutants, indicating an important function for NKCC1 in the maintenance of blood pressure. cAMP-induced short circuit currents, which are dependent on the CFTR Cl(-) channel, were reduced in jejunum, cecum, and trachea of Nkcc1(-/-) mice, indicating that NKCC1 contributes to cAMP-induced Cl(-) secretion. In contrast, secretion of gastric acid in adult Nkcc1(-/-) stomachs and enterotoxin-stimulated fluid secretion in the intestine of suckling Nkcc1(-/-) mice were normal. Finally, homozygous mutants were deaf, and histological analysis of the inner ear revealed a collapse of the membranous labyrinth, consistent with a critical role for NKCC1 in transepithelial K(+) movements involved in generation of the K(+)-rich endolymph and the endocochlear potential. PMID- 10480908 TI - The yeast transcription factor Mac1 binds to DNA in a modular fashion. AB - Mac1 is a metalloregulatory protein that regulates expression of the high affinity copper transport system in the yeast Saccharomyces cerevisiae. Under conditions of high copper concentration, Mac1 represses transcription of genes coding for copper transport proteins. Mac1 binds to DNA sequences called copper response elements (CuREs), which have the consensus sequence 5'-TTTGC(T/G)C(A/G) 3'. Mac1 contains two zinc binding sites, a copper binding site, and the sequence motif RGRP, which has been found in other proteins to mediate binding to the minor groove of A/T-rich sequences in DNA. We have used hydroxyl radical footprinting, missing nucleoside, and methylation interference experiments to investigate the structure of the complex of the DNA binding domain of Mac1 (called here Mac1(t)) with the two CuRE sites found in the yeast CTR1 promoter. We conclude from these experiments that Mac1(t) binds in a modular fashion to DNA, with its RGRP AT-hook motif interacting with the TTT sequence at the 5' end of the CTR1 CuRE site, and with another DNA-binding module(s) binding in the adjacent major groove in the GCTCA sequence. PMID- 10480909 TI - Glypican-6, a new member of the glypican family of cell surface heparan sulfate proteoglycans. AB - The glypicans compose a family of glycosylphosphatidylinositol-anchored heparan sulfate proteoglycans. Mutations in dally, a gene encoding a Drosophila glypican, and in GPC3, the gene for human glypican-3, implicate glypicans in the control of cell growth and division. So far, five members of the glypican family have been identified in vertebrates. By sequencing expressed sequence tag clones and products of rapid amplifications of cDNA ends, we identified a sixth member of the glypican family. The glypican-6 mRNA encodes a protein of 555 amino acids that is most homologous to glypican-4 (identity of 63%). Expression of this protein in Namalwa cells shows a core protein of approximately 60 kDa that is substituted with heparan sulfate only. GPC6, the gene encoding human glypican-6, contains nine exons. Like GPC5, the gene encoding glypican-5, GPC6 maps to chromosome 13q32. Clustering of the GPC5/GPC6 genes on chromosome 13q32 is strongly reminiscent of the clustering of the GPC3/GPC4 genes on chromosome Xq26 and suggests GPCs arose from a series of gene and genome duplications. Based on similarities in sequence and gene organization, glypican-1, glypican-2, glypican 4, and glypican-6 appear to define a subfamily of glypicans, differing from the subfamily comprising so far glypican-3 and glypican-5. Northern blottings indicate that glypican-6 mRNA is widespread, with prominent expressions in human fetal kidney and adult ovary. In situ hybridization studies localize glypican-6 to mesenchymal tissues in the developing mouse embryo. High expressions occur in smooth muscle cells lining the aorta and other major blood vessels and in mesenchymal cells of the intestine, kidney, lung, tooth, and gonad. Growth factor signaling in these tissues might in part be regulated by the presence of glypican 6 on the cell surface. PMID- 10480910 TI - Natural splicing of exon 2 of human interleukin-15 receptor alpha-chain mRNA results in a shortened form with a distinct pattern of expression. AB - We report the existence of eight different interleukin-15 receptor alpha-chain (IL-15Ralpha) transcripts resulting from exon-splicing mechanisms within the IL 15Ralpha gene. Two main classes of transcripts can be distinguished that do or do not (Delta2 isoforms) contain the exon 2-coding sequence. Both classes were expressed in numerous cell lines and tissues (including peripheral blood lymphocytes) at comparable levels and could be transcribed in COS-7 cells, and the proteins were expressed at the cell surface. Both receptor forms displayed numerous glycosylation states, reflecting differential usage of a single N glycosylation site as well as extensive O-glycosylations. Whereas IL-15Ralpha bound IL-15 with high affinity, Delta2IL-15Ralpha was unable to bind IL-15, thus revealing the indispensable role of the exon 2-encoded domain in cytokine binding. A large proportion of IL-15Ralpha was expressed at the nuclear membrane with some intranuclear localization, supporting a potential direct action of the IL-15.IL-15Ralpha complex at the nuclear level. In sharp contrast, Delta2IL 15Ralpha was found only in the non-nuclear membrane compartments, indicating that the exon 2-encoded domain (which is shown to contain a potential nuclear localization signal) plays an important role in receptor post-translational routing. Together, our data indicate that exon 2 splicing of human IL-15Ralpha is a natural process that might play regulatory roles at different levels. PMID- 10480911 TI - Ethanol-induced translocation of cAMP-dependent protein kinase to the nucleus. Mechanism and functional consequences. AB - Ethanol induces translocation of the catalytic subunit (Calpha) of cAMP-dependent protein kinase (PKA) from the Golgi area to the nucleus in NG108-15 cells. Ethanol also induces translocation of the RIIbeta regulatory subunit of PKA to the nucleus; RI and Cbeta are not translocated. Nuclear PKA activity in ethanol treated cells is no longer regulated by cAMP. Gel filtration and immunoprecipitation analysis confirm that ethanol blocks the reassociation of Calpha with RII but does not induce dissociation of these subunits. Ethanol also reduces inhibition of Calpha by the PKA inhibitor PKI. Pre-incubation of Calpha with ethanol decreases phosphorylation of Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide) and casein but has no effect on the phosphorylation of highly charged molecules such as histone H1 or protamine. cAMP-response element-binding protein (CREB) phosphorylation by Calpha is also increased in ethanol-treated cells. This increase in CREB phosphorylation is inhibited by the PKA antagonist (R(p))-cAMPS and by an adenosine receptor antagonist. These results suggest that ethanol affects a cascade of events allowing for sustained nuclear localization of Calpha and prolonged CREB phosphorylation. These events may account for ethanol-induced changes in cAMP-dependent gene expression. PMID- 10480913 TI - Genetic analysis of glutathione peroxidase in oxidative stress response of Saccharomyces cerevisiae. AB - Three glutathione peroxidase homologs (YKL026C, YBR244W, and YIR037W/HYR1) were found in the Saccharomyces Genome Database. We named them GPX1, GPX2, and GPX3, respectively, and we investigated the function of each gene product. The gpx3Delta mutant was hypersensitive to peroxides, whereas null mutants of the GPX1 and GPX2 did not show any obvious phenotypes. Glutathione peroxidase activity decreased approximately 57 and 93% in the gpx3Delta and gpx1Delta/gpx2Delta/gpx3Delta mutants, respectively, compared with that of wild type. Expression of the GPX3 gene was not induced by any stresses tested, whereas that of the GPX1 gene was induced by glucose starvation. The GPX2 gene expression was induced by oxidative stress, which was dependent upon the Yap1p. The TSA1 (thiol-specific antioxidant) gene encodes thioredoxin peroxidase that can reduce peroxides by using thioredoxin as a reducing power. Disruption of the TSA1 gene enhanced the basal expression level of the Yap1p target genes such as GSH1, GLR1, and GPX2 and that resulted in increases of total glutathione level and activities of glutathione reductase and glutathione peroxidase. However, expression of the TSA1 gene did not increase in the gpx1Delta/gpx2Delta/gpx3Delta mutant. Therefore, de novo synthesis and recycling of glutathione were increased in the tsa1Delta mutant to maintain the catalytic cycle of glutathione peroxidase reaction efficiently as a backup system for thioredoxin peroxidase. PMID- 10480912 TI - Distribution of CTP:phosphocholine cytidylyltransferase (CCT) isoforms. Identification of a new CCTbeta splice variant. AB - CTP:phosphocholine cytidylyltransferase is a major regulator of phosphatidylcholine biosynthesis. A single isoform, CCTalpha, has been studied extensively and a second isoform, CCTbeta, was recently identified. We identify and characterize a third cDNA, CCTbeta2, that differs from CCTbeta1 at the carboxyl-terminal end and is predicted to arise as a splice variant of the CCTbeta gene. Like CCTalpha, CCTbeta2 is heavily phosphorylated in vivo, in contrast to CCTbeta1. CCTbeta1 and CCTbeta2 mRNAs were differentially expressed by the human tissues examined, whereas CCTalpha was more uniformly represented. Using isoform-specific antibodies, both CCTbeta1 and CCTbeta2 localized to the endoplasmic reticulum of cells, in contrast to CCTalpha which resided in the nucleus in addition to associating with the endoplasmic reticulum. CCTbeta2 protein has enzymatic activity in vitro and was able to complement the temperature-sensitive cytidylyltransferase defect in CHO58 cells, just as CCTalpha and CCTbeta1 supporting proliferation at the nonpermissive conditions. Overexpression experiments did not reveal discrete physiological functions for the three isoforms that catalyze the same biochemical reaction; however, the differential cellular localization and tissue-specific distribution suggest that CCTbeta1 and CCTbeta2 may play a role that is distinct from ubiquitously expressed CCTalpha. PMID- 10480914 TI - A mutant form of human protein farnesyltransferase exhibits increased resistance to farnesyltransferase inhibitors. AB - Protein farnesyltransferase (FTase) is a key enzyme responsible for the lipid modification of a large and important number of proteins including Ras. Recent demonstrations that inhibitors of this enzyme block the growth of a variety of human tumors point to the importance of this enzyme in human tumor formation. In this paper, we report that a mutant form of human FTase, Y361L, exhibits increased resistance to farnesyltransferase inhibitors, particularly a tricyclic compound, SCH56582, which is a competitive inhibitor of FTase with respect to the CAAX (where C is cysteine, A is an aliphatic amino acid, and X is the C-terminal residue that is preferentially serine, cysteine, methionine, glutamine or alanine) substrates. The Y361L mutant maintains FTase activity toward substrates ending with CIIS. However, the mutant also exhibits an increased affinity for peptides terminating with CIIL, a motif that is recognized by geranylgeranyltransferase I (GGTase I). The Y361L mutant also demonstrates activity with Ha-Ras and Cdc42Hs proteins, substrates of FTase and GGTase I, respectively. In addition, the Y361L mutant shows a marked sensitivity to a zinc chelator HPH-5 suggesting that the mutant has altered zinc coordination. These results demonstrate that a single amino acid change at a residue at the active site can lead to the generation of a mutant resistant to FTase inhibitors. Such a mutant may be valuable for the study of the effects of FTase inhibitors on tumor cells. PMID- 10480915 TI - Identification of three key active site residues in the C-terminal domain of human recombinant folylpoly-gamma-glutamate synthetase by site-directed mutagenesis. AB - Three cysteines in human recombinant folylpoly-gamma-glutamate synthetase (FPGS) that were reactive with iodoacetamide were located in peptides that were highly conserved across species; the functions of two of these peptides, located in the C-terminal domain, were studied by site-directed mutagenesis. When cDNAs containing mutations in each conserved ionic residue on these peptides were transfected into AUXB1 cells, which lack endogenous FPGS activity, one mutant (D335A) did not complement the auxotrophy, and another (R377A) allowed only minimal growth. FPGS activity could not be detected in insect cells expressing abundant levels of these two mutant proteins from recombinant baculoviruses nor from a virus encoding an H338A mutant FPGS. Kinetic analysis of the purified proteins demonstrated that each of these three mutants was quite different from the others. The major kinetic change detected for the H338A mutation was a 600 fold increase in the K(m) for glutamic acid. For the D335A mutation, the binding of all three substrates (aminopterin, ATP, and glutamic acid) was affected. For R377A, the K(m) for glutamic acid was increased by 1500-fold, and there was an approximately 20-fold decrease in the k(cat) of the reaction. The binding of the K(+) ion, a known activator of FPGS, was affected by the D335A and H338A mutations. We conclude that these three amino acids participate in the alignment of glutamic acid in the active site and that Arg-377 is also involved in the mechanism of the reaction. PMID- 10480916 TI - Targeting Janus kinase 3 in mast cells prevents immediate hypersensitivity reactions and anaphylaxis. AB - Janus kinase 3 (JAK3), a member of the Janus family protein-tyrosine kinases, is expressed in mast cells, and its enzymatic activity is enhanced by IgE receptor/FcepsilonRI cross-linking. Selective inhibition of JAK3 in mast cells with 4-(4'-hydroxylphenyl)-amino-6, 7-dimethoxyquinazoline) (WHI-P131) blocked the phospholipase C activation, calcium mobilization, and activation of microtubule-associated protein kinase after lgE receptor/FcepsilonRI cross linking. Treatment of IgE-sensitized rodent as well as human mast cells with WHI P131 effectively inhibited the activation-associated morphological changes, degranulation, and proinflammatory mediator release after specific antigen challenge without affecting the functional integrity of the distal secretory machinery. In vivo administration of the JAK3 inhibitor WHI-P131 prevented mast cell degranulation and development of cutaneous as well as systemic fatal anaphylaxis in mice at nontoxic dose levels. Thus, JAK3 plays a pivotal role in IgE receptor/FcepsilonRI-mediated mast cell responses, and targeting JAK3 with a specific inhibitor, such as WHI-P131, may provide the basis for new and effective treatment as well as prevention programs for mast cell-mediated allergic reactions. PMID- 10480917 TI - Coordinated movement of RACK1 with activated betaIIPKC. AB - Protein kinase C (PKC) isozymes move upon activation from one intracellular site to another. PKC-binding proteins, such as receptors for activated C kinase (RACKs), play an important role in regulating the localization and diverse functions of PKC isozymes. RACK1, the receptor for activated betaIIPKC, determines the localization and functional activity of betaIIPKC. However, the mechanism by which RACK1 localizes activated betaIIPKC is not known. Here, we provide evidence that the intracellular localization of RACK1 changes in response to PKC activation. In Chinese hamster ovary cells transfected with the dopamine D2L receptor and in NG108-15 cells, PKC activation by either phorbol ester or a dopamine D2 receptor agonist caused the movement of RACK1. Moreover, PKC activation resulted in the in situ association and movement of RACK1 and betaIIPKC to the same intracellular sites. Time course studies indicate that PKC activation induces the association of the two proteins prior to their co movement. We further show that association of RACK1 and betaIIPKC is required for the movement of both proteins. Our results suggest that RACK1 is a PKC shuttling protein that moves betaIIPKC from one intracellular site to another. PMID- 10480918 TI - The active site of Escherichia coli UDP-N-acetylglucosamine acyltransferase. Chemical modification and site-directed mutagenesis. AB - UDP-N-acetylglucosamine (UDP-GlcNAc) acyltransferase (LpxA) catalyzes the reversible transfer of an R-3-hydroxyacyl chain from R-3-hydroxyacyl-acyl carrier protein to the glucosamine 3-OH of UDP-GlcNAc in the first step of lipid A biosynthesis. Lipid A is required for the growth and virulence of most Gram negative bacteria, making its biosynthetic enzymes intriguing targets for the development of new antibacterial agents. LpxA is a member of a large family of left-handed beta-helical proteins, many of which are acyl- or acetyltransferases. We now demonstrate that histidine-, lysine-, and arginine-specific reagents effectively inhibit LpxA of Escherichia coli, whereas serine- and cysteine specific reagents do not. Using this information in conjunction with multiple sequence alignments, we constructed site-directed alanine substitution mutations of conserved histidine, lysine, and arginine residues. Many of these mutant LpxA enzymes show severely decreased specific activities under standard assay conditions. The decrease in activity corresponds to decreased k(cat)/K(m,UDP GlcNAc) values for all the mutants. With the exception of H125A, in which no activity is seen under any assay condition, the decrease in k(cat)/K(m,UDP GlcNAc) mainly reflects an increased K(m,UDP-GlcNAc). His(125) of E. coli LpxA may therefore function as a catalytic residue, possibly as a general base. LpxA does not catalyze measurable UDP-3-O-(R-3-hydroxymyristoyl)-GlcNAc hydrolysis or UDP-GlcNAc/UDP-3-O-(R-3-hydroxymyristoyl)-GlcNAc exchange, arguing against a ping pong mechanism with an acyl-enzyme intermediate. PMID- 10480919 TI - Sertolin is a novel gene marker of cell-cell interactions in the rat testis. AB - A novel testicular protein designated sertolin was cloned. The full-length sertolin cDNA consists of 853 base pairs with an open reading frame of 381 base pairs coding for a 127-amino acid polypeptide that shares limited identities with antaxin/josephin and thrombospondin proteins. Sertolin (calculated molecular mass, 13,759 daltons) has two mRNA transcripts of 2.3 and 1 kilobase. A 22-amino acid peptide based on the deduced amino acid sequence of sertolin (NH(2) KKEHFNLFKAASVSHLVQVVPQ) was synthesized and used for polyclonal antibody production. Immunoblot analysis detected a 17-kDa immunoreactive band in the Sertoli cell cytosol. Using Sertoli-germ cell cocultures, sertolin expression was found to be reduced by as much as 5-fold at the time when germ cells attach onto Sertoli cells but preceding the establishment of specialized inter-Sertoli-germ cell junctions. Neither FSH nor 17beta-hydroxy-5alpha-androstan-3-one was able to affect sertolin expression, whereas estradiol-17beta and progesterone induced a significant increase in Sertoli cell sertolin expression in vitro. In addition, interleukin-1alpha, a germ cell-derived cytokine, was also able to elicit a transient but significant increase in Sertoli cell sertolin expression. Sertolin expression was also shown to increase with testicular development and is likely to be associated with the onset of spermatogenesis. In addition, sertolin expression increased in the testis when generalized inflammation was induced in adult rats by injection of fermented yeast. These results show that sertolin will be useful in characterizing cell-cell interactions in the testis. PMID- 10480920 TI - Nitrosothiol formation catalyzed by ceruloplasmin. Implication for cytoprotective mechanism in vivo. AB - Ceruloplasmin (CP) is a major multicopper-containing plasma protein that is not only involved in iron metabolism through its ferroxidase activity but also functions as an antioxidant. However, physiological substrates for CP have not been fully identified nor has the role of CP been fully understood. The reaction of nitric oxide (NO) with CP was investigated in view of nitrosothiol (RS-NO) formation. First, formation of heavy metal- or CP-catalyzed RS-NO was examined with physiologically relevant concentrations of NO and various thiol compounds (RSH) such as glutathione (GSH). Among the various heavy metal ions and copper containing enzymes and proteins examined, only copper ion (Cu(2+)) and CP showed potent RS-NO (S-nitrosoglutathione)-producing activity. Also, RS-NO-forming catalytic activity was evident for CP added exogenously to RAW264 cells expressing inducible NO synthase in culture, but this was not the case for copper ion. Similarly, CP produced endogenously by HepG2 cells showed potent RS-NO forming activity in the cell culture. One-electron oxidation of NO appears to be operative for RS-NO production via electron transfer from type 1 copper to a cluster of types 2 and 3 copper in CP. Neurological disorders are associated with aceruloplasminemia; besides RS-NO, S-nitrosoglutathione particularly has been shown to have neuroprotective effect against oxidative stress induced by iron overload. Thus, we suggest that CP plays an important catalytic role in RS-NO formation, which may contribute to its potent antioxidant and cytoprotective activities in vivo in mammalian biological systems. PMID- 10480921 TI - The kangaroo cation-independent mannose 6-phosphate receptor binds insulin-like growth factor II with low affinity. AB - The mammalian cation-independent mannose 6-phosphate receptor (CI-MPR) binds mannose 6-phosphate-bearing glycoproteins and insulin-like growth factor (IGF) II. However, the CI-MPR from the opossum has been reported to bind bovine IGF-II with low affinity (Dahms, N. M., Brzycki-Wessell, M. A., Ramanujam, K. S., and Seetharam, B. (1993) Endocrinology 133, 440-446). This may reflect the use of a heterologous ligand, or it may represent the intrinsic binding affinity of this receptor. To examine the binding of IGF-II to a marsupial CI-MPR in a homologous system, we have previously purified kangaroo IGF-II (Yandell, C. A., Francis, G. L., Wheldrake, J. F., and Upton, Z. (1998) J. Endocrinol. 156, 195-204), and we now report the purification and characterization of the CI-MPR from kangaroo liver. The interaction of the kangaroo CI-MPR with IGF-II has been examined by ligand blotting, radioreceptor assay, and real-time biomolecular interaction analysis. Using both a heterologous and homologous approach, we have demonstrated that the kangaroo CI-MPR has a lower binding affinity for IGF-II than its eutherian (placental mammal) counterparts. Furthermore, real-time biomolecular interaction analysis revealed that the kangaroo CI-MPR has a higher affinity for kangaroo IGF-II than for human IGF-II. The cDNA sequence of the kangaroo CI-MPR indicates that there is considerable divergence in the area corresponding to the IGF-II binding site of the eutherian receptor. Thus, the acquisition of a high affinity binding site for regulating IGF-II appears to be a recent event specific to the eutherian lineage. PMID- 10480922 TI - Characterization of two isoforms of the skeletal muscle LIM protein 1, SLIM1. Localization of SLIM1 at focal adhesions and the isoform slimmer in the nucleus of myoblasts and cytoplasm of myotubes suggests distinct roles in the cytoskeleton and in nuclear-cytoplasmic communication. AB - We have cloned and characterized a novel isoform of the skeletal muscle LIM protein 1 (SLIM1), designated SLIMMER. SLIM1 contains an N-terminal single zinc finger followed by four LIM domains. SLIMMER is identical to SLIM1 over the first three LIM domains but contains a novel C-terminal 96 amino acids with three potential bipartite nuclear localization signals, a putative nuclear export sequence, and 27 amino acids identical to the RBP-J binding region of KyoT2, a murine isoform of SLIM1. SLIM1 localized to the cytosol of Sol8 myoblasts and myotubes. SLIMMER was detected in the nucleus of myoblasts and, following differentiation into myotubes, was exclusively cytosolic. Recombinant green fluorescent protein-SLIM1 localized to the cytoplasm and associated with focal adhesions and actin filaments in COS-7 cells, while green fluorescent protein SLIMMER was predominantly nuclear. SLIMMER truncation mutants revealed that the first nuclear localization signal mediates nuclear localization. The addition of the proposed nuclear export sequence decreased the level of exclusively nuclear expression and increased cytosolic SLIMMER expression in COS-7 cells. The leucine rich nuclear export signal was required for the export of SLIMMER from the nucleus of myoblasts to the cytoplasm of myotubes. Collectively, these results suggest distinct roles for SLIM1 and SLIMMER in focal adhesions and nuclear cytoplasmic communication. PMID- 10480923 TI - Negative transcriptional regulation mediated by thyroid hormone response element 144 requires binding of the multivalent factor CTCF to a novel target DNA sequence. AB - DNA target sites for a "multivalent" 11-zinc-finger CCTC-binding factor (CTCF) are unusually long ( approximately 50 base pairs) and remarkably different. In conjunction with the thyroid receptor (TR), CTCF binding to the lysozyme gene transcriptional silencer mediates the thyroid hormone response element (TRE) dependent transcriptional repression. We tested whether other TREs, which in addition to the presence of a TR binding site require neighboring sequences for transcriptional function, might also contain a previously unrecognized binding site(s) for CTCF. One such candidate DNA region, previously isolated by Bigler and Eisenman (Bigler, J., and Eisenman, R. N. (1995) EMBO J. 14, 5710-5723), is the TRE-containing genomic element 144. We have identified a new CTCF target sequence that is adjacent to the TR binding site within the 144 fragment. Comparison of CTCF recognition nucleotides in the lysozyme silencer and in the 144 sequences revealed both similarities and differences. Several C-terminal CTCF zinc fingers contribute differently to binding each of these sequences. Mutations that eliminate CTCF binding impair 144-mediated negative transcriptional regulation. Thus, the 144 element provides an additional example of a functionally significant composite "TRE plus CTCF binding site" regulatory element suggesting an important role for CTCF in cooperation with the steroid/thyroid superfamily of nuclear receptors to mediate TRE-dependent transcriptional repression. PMID- 10480924 TI - ADP-ribosylation factor 6 as a target of guanine nucleotide exchange factor GRP1. AB - The GRP1 protein contains a Sec7 homology domain that catalyzes guanine nucleotide exchange on ADP-ribosylation factors (ARF) 1 and 5 as well as a pleckstrin homology domain that binds phosphatidylinositol(3,4,5)P(3), an intermediate in cell signaling by insulin and other extracellular stimuli (Klarlund, J. K., Guilherme, A., Holik, J. J., Virbasius, J. V., Chawla, A., and Czech, M. P. (1997) Science 275, 1927-1930). Here we show that both endogenous GRP1 and ARF6 rapidly co-localize in plasma membrane ruffles in Chinese hamster ovary (CHO-T) cells expressing human insulin receptors and COS-1 cells in response to insulin and epidermal growth factor, respectively. The pleckstrin homology domain of GRP1 appears to be sufficient for regulated membrane localization. Using a novel method to estimate GTP loading of expressed HA epitope-tagged ARF proteins in intact cells, levels of biologically active, GTP bound ARF6 as well as GTP-bound ARF1 were elevated when these ARF proteins were co-expressed with GRP1 or the related protein cytohesin-1. GTP loading of ARF6 in both control cells and in response to GRP1 or cytohesin-1 was insensitive to brefeldin A, consistent with previous data on endogenous ARF6 exchange activity. The ability of GRP1 to catalyze GTP/GDP exchange on ARF6 was confirmed using recombinant proteins in a cell-free system. Taken together, these results suggest that phosphatidylinositol(3,4,5)P(3) may be generated in cell membrane ruffles where receptor tyrosine kinases are concentrated in response to growth factors, causing recruitment of endogenous GRP1. Further, co-localization of GRP1 with ARF6, combined with its demonstrated ability to activate ARF6, suggests a physiological role for GRP1 in regulating ARF6 functions. PMID- 10480925 TI - Purification and characterization of phosphopantetheine adenylyltransferase from Escherichia coli. AB - Phosphopantetheine adenylyltransferase (PPAT) catalyzes the penultimate step in coenzyme A (CoA) biosynthesis: the reversible adenylation of 4' phosphopantetheine yielding 3'-dephospho-CoA and pyrophosphate. Wild-type PPAT from Escherichia coli was purified to homogeneity. N-terminal sequence analysis revealed that the enzyme is encoded by a gene designated kdtB, purported to encode a protein involved in lipopolysaccharide core biosynthesis. The gene, here renamed coaD, is found in a wide range of microorganisms, indicating that it plays a key role in the synthesis of 3'-dephospho-CoA. Overexpression of coaD yielded highly purified recombinant PPAT, which is a homohexamer of 108 kDa. Not less than 50% of the purified enzyme was found to be associated with CoA, and a method was developed for its removal. A steady state kinetic analysis of the reverse reaction revealed that the mechanism of PPAT involves a ternary complex of enzyme and substrates. Since purified PPAT lacks dephospho-CoA kinase activity, the two final steps of CoA biosynthesis in E. coli must be catalyzed by separate enzymes. PMID- 10480926 TI - MutS recognition of exocyclic DNA adducts that are endogenous products of lipid oxidation. AB - The ability of the methyl-directed mismatch repair system to recognize and repair the exocyclic adducts propanodeoxyguanosine (PdG) and pyrimido[1,2-alpha]purin 10(3H)-one (M(1)G), the major adduct derived from the endogenous mutagen malondialdehyde, has been assessed both in vivo and in vitro. Both adducts were site-specifically incorporated into M13MB102 DNA, and the adducted genomes were electroporated into wild-type or mutS-deficient Escherichia coli strains. A decrease in mutations caused by both adducts was observed in mutS-deficient strains, suggesting that MutS was binding to the adducts and blocking repair by nucleotide excision repair. This hypothesis was supported by the differences in mutation frequency observed when hemimethylated genomes containing PdG on the (-) strand were electroporated into a uvrA(-) strain. The ability of purified MutS to bind to PdG- or M(1)G-containing 31-mer duplexes in vitro was assessed using both surface plasmon resonance and gel shift assays. MutS bound to M(1)G:T-containing duplexes with similar affinity to a G:T mismatch but less strongly to M(1)G:C- and PdG-containing duplexes. Dissociation from each of the adduct-containing duplexes occurred at a faster rate than from a G:T mismatch. The present results indicate that MutS can bind to exocyclic adducts resulting from endogenous DNA damage and trigger their removal by mismatch repair or protect them from removal by nucleotide excision repair. PMID- 10480927 TI - Quantitative relationship among integrin-ligand binding, adhesion, and signaling via focal adhesion kinase and extracellular signal-regulated kinase 2. AB - Because integrin-mediated signals are transferred through a physical architecture and synergistic biochemical network whose properties are not well defined, quantitative relationships between extracellular integrin-ligand binding events and key intracellular responses are poorly understood. We begin to address this by quantifying integrin-mediated FAK and ERK2 responses in CHO cells for varied alpha(5)beta(1) expression level and substratum fibronectin density. Plating cells on fibronectin-coated surfaces initiated a transient, biphasic ERK2 response, the magnitude and kinetics of which depended on integrin-ligand binding properties. Whereas ERK2 activity initially increased with a rate proportional to integrin-ligand bond number for low fibronectin density, the desensitization rate was independent of integrin and fibronectin amount but proportional to the ERK2 activity level with an exponential decay constant of 0.3 (+/- 0.08) min(-1). Unlike the ERK2 activation time course, FAK phosphorylation followed a superficially disparate time course. However, analysis of the early kinetics of the two signals revealed them to be correlated. The initial rates of FAK and ERK2 signal generation exhibited similar dependence on fibronectin surface density, with both rates monotonically increasing with fibronectin amount until saturating at high fibronectin density. Because of this similar initial rate dependence on integrin-ligand bond formation, the disparity in their time courses is attributed to differences in feedback regulation of these signals. Whereas FAK phosphorylation increased to a steady-state level as new integrin-ligand bond formation continued during cell spreading, ERK2 activity was decoupled from the integrin-ligand stimulus and decayed back to a basal level. Accordingly, we propose different functional metrics for representing these two disparate dynamic signals: the steady-state tyrosine phosphorylation level for FAK and the integral of the pulse response for ERK2. These measures of FAK and ERK2 activity were found to correlate with short term cell-substratum adhesivity, indicating that signaling via FAK and ERK2 is proportional to the number of integrin-fibronectin bonds. PMID- 10480928 TI - Selective nucleosome disruption by drugs that bind in the minor groove of DNA. AB - Previous studies have shown that drugs which bind in the DNA minor groove reduce the curvature of bent DNA. In this article, we examined the effects of these drugs on the nucleosome assembly of DNA molecules that display different degrees of intrinsic curvature. DAPI (4,6-diamidino-2-phenylindole) inhibited the assembly of a histone octamer onto a 192-base pair circular DNA fragment from Caenorhabditis elegans and destabilized a nucleosome that was previously assembled on this segment. The inhibitory effect was highly selective since it was not seen with nonbent molecules, bent molecules with noncircular shapes, or total genomic DNA. This marked template specificity was attributed to the binding of the ligand to multiple oligo A-tracts distributed over the length of the fragment. A likely mechanism for the effect is that the bound ligand prevents the further compression of the DNA into the minor groove which is required for assembly of DNA into nucleosomes. To further characterize the effects of the drug on chromatin formation, a nucleosome was assembled onto a 322-base pair DNA fragment that contained the circular element and a flanking nonbent segment of DNA. The position of the nucleosome along the fragment was then determined using a variety of nuclease probes including exonuclease III, micrococcal nuclease, DNase I, and restriction enzymes. The results of these studies revealed that the nucleosome was preferentially positioned along the circular element in the absence of DAPI but assembled onto the nonbent flanking sequence in the presence of the drug. DAPI also induced the directional movement of the nucleosome from the circular element onto the nonbent flanking sequence when a nucleosome preassembled onto this template was exposed to the drug under physiologically relevant conditions. PMID- 10480929 TI - A single mutation, RecB(D1080A,) eliminates RecA protein loading but not Chi recognition by RecBCD enzyme. AB - Homologous recombination and double-stranded DNA break repair in Escherichia coli are initiated by the multifunctional RecBCD enzyme. After binding to a double stranded DNA end, the RecBCD enzyme unwinds and degrades the DNA processively. This processing is regulated by the recombination hot spot, Chi (chi: 5'-GCTGGTGG 3'), which induces a switch in the polarity of DNA degradation and activates RecBCD enzyme to coordinate the loading of the DNA strand exchange protein, RecA, onto the single-stranded DNA products of unwinding. Recently, a single mutation in RecB, Asp-1080 --> Ala, was shown to create an enzyme (RecB(D1080A)CD) that is a processive helicase but not a nuclease. Here we show that the RecB(D1080A)CD enzyme is also unable to coordinate the loading of the RecA protein, regardless of whether chi sites are present in the DNA. However, the RecB(D1080A)CD enzyme does respond to chi sites by inactivating in a chi-dependent manner. These data define a locus of the RecBCD enzyme that is essential not only for nuclease function but also for the coordination of RecA protein loading. PMID- 10480930 TI - Up-regulation of cell-surface alpha4beta2 neuronal nicotinic receptors by lower temperature and expression of chimeric subunits. AB - The predominant nicotinic acetylcholine receptor (nAChR) expressed in vertebrate brain is a pentamer containing alpha4 and beta2 subunits. In this study we have examined how temperature and the expression of subunit chimeras can influence the efficiency of cell-surface expression of the rat alpha4beta2 nAChR. Functional recombinant alpha4beta2 nAChRs, showing high affinity binding of nicotinic radioligands (K(d) = 41 +/- 22 pM for [(3)H]epibatidine), are expressed in both stably and transiently transfected mammalian cell lines. Despite this, only very low levels of alpha4beta2 nAChRs can be detected on the cell surface of transfected mammalian cells maintained at 37 degrees C. At 30 degrees C, however, cells expressing alpha4beta2 nAChRs show a 12-fold increase in radioligand binding (with no change in affinity), and a 5-fold up-regulation in cell-surface receptors with no increase in total subunit protein. In contrast to "wild-type" alpha4 and beta2 subunits, chimeric nicotinic/serotonergic subunits ("alpha4chi" and "beta2chi") are expressed very efficiently on the cell surface (at 30 degrees C or 37 degrees C), either as hetero-oligomeric complexes (e.g. alpha4chi+beta2 or alpha4chi+beta2chi) or when expressed alone. Compared with alpha4beta2 nAChRs, expression of complexes containing chimeric subunits typically results in up to 20-fold increase in nicotinic radioligand binding sites (with no change in affinity) and a similar increase in cell-surface receptor, despite a similar level of total chimeric and wild-type protein. PMID- 10480931 TI - Homing in on the role of transition metals in the HNH motif of colicin endonucleases. AB - The cytotoxic domain of the bacteriocin colicin E9 (the E9 DNase) is a nonspecific endonuclease that must traverse two membranes to reach its cellular target, bacterial DNA. Recent structural studies revealed that the active site of colicin DNases encompasses the HNH motif found in homing endonucleases, and bound within this motif a single transition metal ion (either Zn(2+) or Ni(2+)) the role of which is unknown. In the present work we find that neither Zn(2+) nor Ni(2+) is required for DNase activity, which instead requires Mg(2+) ions, but binding transition metals to the E9 DNase causes subtle changes to both secondary and tertiary structure. Spectroscopic, proteolytic, and calorimetric data show that, accompanying the binding of 1 eq of Zn(2+), Ni(2+), or Co(2+), the thermodynamic stability of the domain increased substantially, and that the equilibrium dissociation constant for Zn(2+) was less than or equal to nanomolar, while that for Co(2+) and Ni (2+) was micromolar. Our data demonstrate that the transition metal is not essential for colicin DNase activity but rather serves a structural role. We speculate that the HNH motif has been adapted for use by endonuclease colicins because of its involvement in DNA recognition and because removal of the bound metal ion destabilizes the DNase domain, a likely prerequisite for its translocation across bacterial membranes. PMID- 10480932 TI - Involvement of the p38 mitogen-activated protein kinase pathway in transforming growth factor-beta-induced gene expression. AB - Transforming growth factor-beta (TGF-beta)-activated kinase 1 (TAK1), a member of the mitogen-activated protein kinase kinase kinase family, is suggested to be involved in TGF-beta-induced gene expression, but the signaling mechanism from TAK1 to the nucleus remains largely undefined. We have found that p38 mitogen activated protein kinase, and its direct activator MKK6 are rapidly activated in response to TGF-beta. Expression of dominant negative MKK6 or dominant negative TAK1 inhibited the TGF-beta-induced transcriptional activation as well as the p38 activation. Constitutive activation of the p38 pathway in the absence of TGF-beta induced the transcriptional activation, which was enhanced synergistically by coexpression of Smad2 and Smad4 and was inhibited by expression of the C-terminal truncated, dominant negative Smad4. Furthermore, we have found that activating transcription factor-2 (ATF-2), which is known as a nuclear target of p38, becomes phosphorylated in the N-terminal activation domain in response to TGF beta, that ATF-2 forms a complex with Smad4, and that the complex formation is enhanced by TGF-beta. In addition, expression of a nonphosphorylatable form of ATF-2 inhibited the TGF-beta-induced transcriptional activation. These results show that the p38 pathway is activated by TGF-beta and is involved in the TGF beta-induced transcriptional activation by regulating the Smad-mediated pathway. PMID- 10480933 TI - 90-kDa ribosomal S6 kinase is phosphorylated and activated by 3-phosphoinositide dependent protein kinase-1. AB - 90-kDa ribosomal S6 kinase-2 (RSK2) belongs to a family of growth factor activated serine/threonine kinases composed of two kinase domains connected by a regulatory linker region. The N-terminal kinase of RSK2 is involved in substrate phosphorylation. Its activation requires phosphorylation of the linker region at Ser(369), catalyzed by extracellular signal-regulated kinase (ERK), and at Ser(386), catalyzed by the C-terminal kinase, after its activation by ERK. In addition, the N-terminal kinase must be phosphorylated at Ser(227) in the activation loop by an as yet unidentified kinase. Here, we show that the isolated N-terminal kinase of RSK2 (amino acids 1-360) is phosphorylated at Ser(227) by PDK1, a constitutively active kinase, leading to 100-fold stimulation of kinase activity. In COS7 cells, ectopic PDK1 induced the phosphorylation of full-length RSK2 at Ser(227) and Ser(386), without involvement of ERK, leading to partial activation of RSK2. Similarly, two other members of the RSK family, RSK1 and RSK3, were partially activated by PDK1 in COS7 cells. Finally, our data indicate that full activation of RSK2 by growth factor requires the cooperation of ERK and PDK1 through phosphorylation of Ser(227), Ser(369), and Ser(386). Our study extend recent findings which implicate PDK1 in the activation of protein kinases B and C and p70(S6K), suggesting that PDK1 controls several major growth factor activated signal transduction pathways. PMID- 10480935 TI - Molecular characterization of the PmrA regulon. AB - The two-component system PmrA/PmrB of Salmonella enterica controls expression of several loci including those mediating modifications in the lipopolysaccharide that result in polymyxin resistance. To gain insight in the regulation of polymyxin resistance, we mapped the transcription start sites of the PmrA regulated genes pmrC, pmrG, pbgPE, and ugd and identified a conserved sequence in the promoter region of the first three genes. His-tagged PmrA protein could gel shift DNA fragments containing the promoters of the pmrC, pmrG, and pbgPE genes but not the udg promoter. DNase I footprinting analysis of the pmrC, pmrG, and pbgPE promoters indicate that phosphorylated as well as unphosphorylated PmrA bind to a 16-base pair imperfect inverted repeat sequence (5'-TTAAKTTCTTAAKGTT 3'), which is found 40, 80, and 38 nucleotides upstream from the transcription start sites of the pmrC, pmrG, and pbgPE genes, respectively. Our data suggest that a PmrA dimer activates transcription of the divergent pmrG and pbgPE promoters by binding to a single site in the pmrG-pbgPE intergenic region and that the ugd gene is regulated by the PmrA/PmrB system only indirectly. PMID- 10480934 TI - c-Raf-mediated inhibition of epidermal growth factor-stimulated cell migration. AB - Epidermal growth factor stimulates migration of a number of cell types, yet the signaling pathways that regulate epidermal growth factor-stimulated migration are poorly defined. In this report, we employ a transient transfection migration assay to assess the role of components of the Ras-mitogen-activated protein (MAP) kinase signaling pathway in epidermal growth factor-stimulated chemotaxis of rat embryo fibroblasts. Expression of dominant negative Ras blocks epidermal growth factor-mediated chemotaxis, while constitutively active Ras has no effect on chemokinesis or chemotaxis. PD98059 and U0126, inhibitors of MAP kinase kinase (MEK) activity, decreased epidermal growth factor-stimulated migration, while kinase-defective MEK1, an inhibitor of MAP kinase activation, enhanced migration. To understand the paradoxical effects of these molecules on epidermal growth factor-induced migration, we examined the role of c-Raf on migration. Expression of either wild type c-Raf or the catalytic domain of c-Raf effectively inhibited epidermal growth factor-stimulated cell migration. We suggest that, whereas Ras activity is necessary to promote epidermal growth factor-stimulated migration, sustained activation of c-Raf may be important in down-regulating migratory signaling pathways triggered by epidermal growth factor receptor activation. Further, activation of c-Raf upon inhibition of the MEK-MAP kinase pathway may contribute to the inhibition of cell migration observed with pharmacological MEK inhibitors. PMID- 10480936 TI - Generation of a novel A kinase anchor protein and a myristoylated alanine-rich C kinase substrate-like analog from a single gene. AB - A unique Drosophila gene encodes two novel signaling proteins. Drosophila A kinase anchor protein 200 (DAKAP200) (753 amino acids) binds regulatory subunits of protein kinase AII (PKAII) isoforms in vitro and in intact cells. The acidic DAKAP200 polypeptide (pI approximately 3.8) contains an optimal N-terminal myristoylation site and a positively charged domain that resembles the multifunctional phosphorylation site domain of vertebrate myristoylated alanine rich C kinase substrate proteins. The 15-kilobase pair DAKAP200 gene contains six exons and encodes a second protein, DeltaDAKAP200. DeltaDAKAP200 is derived from DAKAP200 transcripts by excision of exon 5 (381 codons), which encodes the PKAII binding region and a Pro-rich sequence. DeltaDAKAP200 appears to be a myristoylated alanine-rich C kinase substrate analog. DAKAP200 and DeltaDAKAP200 are evident in vivo at all stages of Drosophila development. Thus, both proteins may play important physiological roles throughout the life span of the organism. Nevertheless, DAKAP200 gene expression is regulated. Maximal levels of DAKAP200 are detected in the pupal phase of development; DeltaDAKAP200 content is elevated 7-fold in adult head (brain) relative to other body parts. Enhancement or suppression of exon 5 excision during DAKAP200 pre-mRNA processing provides potential mechanisms for regulating anchoring of PKAII and targeting of cAMP signals to effector sites in cytoskeleton and/or organelles. PMID- 10480937 TI - Characterization of the targeting, binding, and phosphorylation site domains of an A kinase anchor protein and a myristoylated alanine-rich C kinase substrate like analog that are encoded by a single gene. AB - A novel Drosophila A kinase anchor protein, Drosophila A kinase anchor protein 200 (DAKAP200), is predicted to be involved in routing, mediating, and integrating signals carried by cAMP, Ca(2+), and diacylglycerol (Li, Z., Rossi, E. A., Hoheisel, J. D., Kalderon, D., and Rubin, C. S. (1999) J. Biol. Chem. 274, 27191-27200). Experiments designed to assess this hypothesis now (a) establish the function, boundaries and identity of critical amino acids of the protein kinase AII (PKAII) tethering site of DAKAP200; (b) demonstrate that residues 119 148 mediate binding with Ca(2+)-calmodulin and F-actin; (c) show that a polybasic region of DAKAP200 is a substrate for protein kinase C; (d) reveal that phosphorylation of the polybasic domain regulates affinity for F-actin and Ca(2+) calmodulin; and (e) indicate that DAKAP200 is myristoylated and that this modification promotes targeting of DAKAP200 to plasma membrane. DeltaDAKAP200, a second product of the DAKAP200 gene, cannot tether PKAII. However, DeltaDAKAP200 is myristoylated and contains a phosphorylation site domain that binds Ca(2+) calmodulin and F-actin. An atypical amino acid composition, a high level of negative charge, exceptional thermostability, unusual hydrodynamic properties, properties of the phosphorylation site domain, and a calculated M(r) of 38,000 suggest that DeltaDAKAP200 is a new member of the myristoylated alanine-rich C kinase substrate protein family. DAKAP200 is a potentially mobile, chimeric A kinase anchor protein-myristoylated alanine-rich C kinase substrate protein that may facilitate localized reception and targeted transmission of signals carried by cAMP, Ca(2+), and diacylglycerol. PMID- 10480938 TI - Biochemical evidence for the co-association of three N-methyl-D-aspartate (NMDA) R2 subunits in recombinant NMDA receptors. AB - Functional characterization of wild-type and mutant cloned N-methyl-D-aspartate (NMDA) receptors has been used to deduce their subunit stoichiometry and quaternary structure. However, the results reported from different groups have been at variance and are thus inconclusive. This study has employed a biochemical approach to determine the number of NMDA R2 (NR2) subunits/receptor together with the NMDA R1 (NR1)/NR2 subunit ratio of both cloned and native NMDA receptors. Thus, human embryonic kidney 293 cells were transfected with the NR1-1a and NR2A NMDA receptor subunits in combination with both FLAG- and c-Myc epitope-tagged NR2B subunits. The expressed receptors were detergent-extracted and subjected to double immunoaffinity purification using anti-NR2A and anti-FLAG antibody immunoaffinity columns in series. Immunoblotting of the double immunopurified NR2A/NR2B(FLAG)-containing material demonstrated the presence of anti-NR1, anti NR2A, anti-FLAG, and, more important, anti-c-Myc antibody immunoreactivities. The presence of anti-c-Myc antibody immunoreactivity in the double immunoaffinity purified material showed the co-assembly of three NR2 subunits, i.e. NR2A/NR2B(FLAG)/NR2B(c-Myc), within the same NMDA receptor complex. Control experiments excluded the possibility that the co-immunopurification of the three NR2 subunits was an artifact of the solubilization procedure. These results, taken together with those previously described that showed two NR1 subunits/oligomer, suggest that the NMDA receptor is at least pentameric. PMID- 10480939 TI - Chloroplast FtsY, chloroplast signal recognition particle, and GTP are required to reconstitute the soluble phase of light-harvesting chlorophyll protein transport into thylakoid membranes. AB - The integration of light-harvesting chlorophyll proteins (LHCPs) into the thylakoid membrane proceeds in two steps. First, LHCP interacts with a chloroplast signal recognition particle (cpSRP) to form a soluble targeting intermediate called the transit complex. Second, LHCP integrates into the thylakoid membrane in the presence of GTP, at least one other soluble factor, and undefined membrane components. We previously determined that cpSRP is composed of 43- and 54-kDa polypeptides. We have examined the subunit stoichiometry of cpSRP and find that it is trimeric and composed of two subunits of cpSRP43/subunit of cpSRP54. A chloroplast homologue of FtsY, an Escherichia coli protein that is critical for the function of E. coli SRP, was found largely in the stroma unassociated with cpSRP. When chloroplast FtsY was combined with cpSRP and GTP, the three factors promoted efficient LHCP integration into thylakoid membranes in the absence of stroma, demonstrating that they are all required for reconstituting the soluble phase of LHCP transport. PMID- 10480940 TI - Wiskott-Aldrich syndrome protein induces actin clustering without direct binding to Cdc42. AB - WASP (Wiskott-Aldrich syndrome protein) was identified as the gene product whose mutation causes the human hereditary disease Wiskott-Aldrich syndrome. WASP contains many functional domains and has been shown to induce the formation of clusters of actin filaments in a manner dependent on Cdc42. However, there has been no report investigating what domain(s) is(are) important for the function. Here we present for the first time the results of detailed analyses on the domain function relationship of WASP. First, the C-terminal verprolin-cofilin-acidic domain was shown to be essential for the regulation of actin cytoskeleton. In addition, we found that the clustering of WASP itself is distinct from actin clustering. The partial protein containing the region from the N-terminal pleckstrin homology domain to the basic residue-rich region also clustered especially around the nucleus as wild type WASP without inducing actin clustering. Finally, we obtained the quite unexpected result that a WASP mutant deficient in binding to Cdc42 still induced actin cluster formation, indicating that direct interaction between Cdc42 and WASP is not required for the regulation of actin cytoskeleton. This result may explain why no Wiskott-Aldrich syndrome patients have been identified with a missense mutation in the Cdc42-binding site. PMID- 10480941 TI - A novel motif mediates the targeting of the Arabidopsis COP1 protein to subnuclear foci. AB - The constitutive photomorphogenesis 1 (COP1) protein of Arabidopsis thaliana accumulates in discrete subnuclear foci. To better understand the role of subnuclear architecture in COP1-mediated gene expression, we investigated the structural motifs of COP1 that mediate its localization to subnuclear foci using mutational analysis with green fluorescent protein as a reporter. In a transient expression assay, a subnuclear localization signal consisting of 58 residues between amino acids 120 and 177 of COP1 was able to confer speckled localization onto the heterologous nuclear NIa protein from tobacco etch virus. The subnuclear localization signal overlaps two previously characterized motifs, a cytoplasmic localization signal and a putative alpha-helical coiled-coil domain that has been implicated in COP1 dimerization. Moreover, phenotypically lethal mutations in the carboxyl-terminal WD-40 repeats inhibited localization to subnuclear foci, consistent with a functional role for the accumulation of COP1 at subnuclear sites. PMID- 10480942 TI - Expression, purification, and functional analysis of murine ectodomain fragments of CD8alphaalpha and CD8alphabeta dimers. AB - Soluble mouse CD8alphaalpha and CD8alphabeta dimers corresponding to the paired ectodomains (CD8(f)) or their respective component Ig-like domains (CD8) were expressed in Chinese hamster ovary cells or the glycosylation variant Lec3.2.8.1 cells as secreted proteins using a leucine zipper strategy. The affinity of CD8alphaalpha(f) for H-2K(b) as measured by BIAcore revealed a approximately 65 microM K(d), similar to that of CD8alphabeta(f). Consistent with this result, CD8alphaalpha(f) as well as CD8alphabeta(f) blocked the effector function of N15 T cell receptor transgenic cytolytic T cells in a comparable, dose-dependent fashion. Furthermore, both Lec3.2.8.1-produced and Chinese hamster ovary-produced CD8 homodimers and heterodimers were active in the inhibition assay. These results suggest that the Ig-like domains of CD8 molecules are themselves sufficient to block the requisite transmembrane CD8-pMHC interaction between cytolytic T lymphocytes and target cells. Moreover, given the similarities in co receptor affinities for pMHC, the findings suggest that the greater efficiency of CD8alphabeta versus CD8alphaalpha co-receptor function on T cells is linked to differences within their membrane-bound stalk regions and/or intracellular segments. As recently shown for sCD8alphaalpha, the yield, purity and homogeneity of the deglycosylated protein resulting from this expression system is sufficient for crystallization and x-ray diffraction at atomic resolution. PMID- 10480943 TI - Initiation of bidirectional replication at the chromosomal origin is directed by the interaction between helicase and primase. AB - Several protein-protein interactions have been shown to be critical for proper replication fork function in Escherichia coli. These include interactions between the polymerase and the helicase, the helicase and the primase, and the primase and the polymerase. We have studied the influence of these interactions on proper initiation at oriC by using mutant primases defective in their interaction with the helicase and DNA polymerase III holoenzyme lacking the tau subunit so that it will not interact with the helicase. We show here that accurate initiation of bidirectional DNA replication from oriC is dependent on proper placement of the primers for leading strand synthesis and is thus governed primarily by the interaction between the helicase and primase. PMID- 10480944 TI - Amyloid precursor-like protein 2 promotes cell migration toward fibronectin and collagen IV. AB - Previous studies have established that in response to wounding, the expression of amyloid precursor-like protein 2 (APLP2) in the basal cells of migrating corneal epithelium is greatly up-regulated. To further our understanding of the functional significance of APLP2 in wound healing, we have measured the migratory response of transfected Chinese hamster ovary (CHO) cells expressing APLP2 isoforms to a variety of extracellular matrix components including laminin, collagen types I, IV, and VII, fibronectin, and heparan sulfate proteoglycans (HSPGs). CHO cells overexpressing either of two APLP2 variants, differing in chondroitin sulfate (CS) attachment, exhibit a marked increase in chemotaxis toward type IV collagen and fibronectin but not to laminin, collagen types I and VII, and HSPGs. Cells overexpressing APLP2-751 (CS-modified) exhibited a greater migratory response to fibronectin and type IV collagen than their non-CS-attached counterparts (APLP2-763), suggesting that CS modification enhanced APLP2 effects on cell migration. Moreover, in the presence of chondroitin sulfate, transfectants overexpressing APLP2-751 failed to exhibit this enhanced migration toward fibronectin. The APLP2-ECM interactions were also explored by solid phase adhesion assays. While overexpression of APLP2 isoforms moderately enhanced CHO adhesion to laminin, collagen types I and VII, and HSPGs lines, especially those overexpressing APLP2-751, exhibited greatly increased adhesion to type IV collagen and fibronectin. These observations suggest that APLP2 contributes to re epithelialization during wound healing by supporting epithelial cell adhesion to fibronectin and collagen IV, thus influencing their capacity to migrate over the wound bed. Furthermore, APLP2 interactions with fibronectin and collagen IV appear to be potentiated by the addition of a CS chain to the core proteins. PMID- 10480945 TI - Inhibition of fibronectin matrix assembly by the heparin-binding domain of vitronectin. AB - The deposition of fibronectin into the extracellular matrix is an integrin dependent, multistep process that is tightly regulated in order to ensure controlled matrix deposition. Reduced fibronectin deposition has been associated with altered embryonic development, tumor cell invasion, and abnormal wound repair. In one of the initial steps of fibronectin matrix assembly, the amino terminal region of fibronectin binds to cell surface receptors, termed matrix assembly sites. The present study was undertaken to investigate the role of extracellular signals in the regulation of fibronectin deposition. Our data indicate that the interaction of cells with the extracellular glycoprotein, vitronectin, specifically inhibits matrix assembly site expression and fibronectin deposition. The region of vitronectin responsible for the inhibition of fibronectin deposition was localized to the heparin-binding domain. Vitronectin's heparin-binding domain inhibited both beta(1) and non-beta(1) integrin-dependent matrix assembly site expression and could be overcome by treatment of cells with lysophosphatidic acid, an agent that promotes actin polymerization. The interaction of cells with the heparin-binding domain of vitronectin resulted in changes in actin microfilament organization and the subcellular distribution of the actin-associated proteins alpha-actinin and talin. These data suggest a mechanism whereby the heparin-binding domain of vitronectin regulates the deposition of fibronectin into the extracellular matrix through alterations in the organization of the actin cytoskeleton. PMID- 10480946 TI - Myosin II folding is mediated by a molecular chaperonin. AB - The folding pathway of the heavy meromyosin subfragment (HMM) of a skeletal muscle myosin has been investigated by in vitro synthesis of the myosin heavy and light chains in a coupled transcription and translation assay. Analysis of the nascent translation products for folding intermediates has identified a major intermediate that contains all three myosin subunits in a complex with the eukaryotic cytosolic chaperonin. Partially folded HMM is released from this complex in an ATP-dependent manner. However, biochemical and functional assays reveal incomplete folding of the myosin motor domain. Dimerization of myosin heavy chains and association of heavy and light chains are accomplished early in the folding pathway. To test for other factors necessary for the complete folding of myosin, a cytoplasmic extract was prepared from myotubes produced by a mouse myogenic cell line. This extract dramatically enhanced the folding of HMM, suggesting a role for muscle-specific factors in the folding pathway. We conclude that the molecular assembly of myosin is mediated by the eukaryotic cytosolic chaperonin with folding of the motor domain as the slow step in the pathway. PMID- 10480947 TI - Clostridium septicum alpha toxin uses glycosylphosphatidylinositol-anchored protein receptors. AB - The alpha toxin produced by Clostridium septicum is a channel-forming protein that is an important contributor to the virulence of the organism. Chinese hamster ovary (CHO) cells are sensitive to low concentrations of the toxin, indicating that they contain toxin receptors. Using retroviral mutagenesis, a mutant CHO line (BAG15) was generated that is resistant to alpha toxin. FACS analysis showed that the mutant cells have lost the ability to bind the toxin, indicating that they lack an alpha toxin receptor. The mutant cells are also resistant to aerolysin, a channel-forming protein secreted by Aeromonas spp., which is structurally and functionally related to alpha toxin and which is known to bind to glycosylphosphatidylinositol (GPI)-anchored proteins, such as Thy-1. We obtained evidence that the BAG15 cells lack N-acetylglucosaminyl phosphatidylinositol deacetylase-L, needed for the second step in GPI anchor biosynthesis. Several lymphocyte cell lines lacking GPI-anchored proteins were also shown to be less sensitive to alpha toxin. On the other hand, the sensitivity of CHO cells to alpha toxin was increased when the cells were transfected with the GPI-anchored folate receptor. We conclude that alpha toxin, like aerolysin, binds to GPI-anchored protein receptors. Evidence is also presented that the two toxins bind to different subsets of GPI-anchored proteins. PMID- 10480948 TI - Number of subunits comprising the epithelial sodium channel. AB - The human epithelial sodium channel (hENaC) is a hetero-oligomeric complex composed of three subunits, alpha, beta, and gamma. Understanding the structure and function of this channel and its abnormal behavior in disease requires knowledge of the number of subunits that comprise the channel complex. We used freeze-fracture electron microscopy and electrophysiological methods to evaluate the number of subunits in the ENaC complex expressed in Xenopus laevis oocytes. In oocytes expressing wild-type hENaC (alpha, beta, and gamma subunits), clusters of particles appeared in the protoplasmic face of the plasma membrane. The total number of particles in the clusters was consistent with the whole-cell amiloride sensitive current measured in the same cells. The size frequency histogram for the particles in the clusters suggested the presence of an integral membrane protein complex composed of 17 +/- 2 transmembrane alpha-helices. Because each ENaC subunit has two putative transmembrane helices, these data suggest that in the oocyte plasma membrane, the ENaC complex is composed of eight or nine subunits. At high magnification, individual ENaC particles exhibited a near square geometry. Functional studies using wild-type alphabeta-hENaC coexpressed with gamma-hENaC mutants, which rendered the functional channel differentially sensitive to methanethiosulfonate reagents and cadmium, suggested that the functional channel complex contains more than one gamma subunit. These data suggest that functional ENaC consists of eight or nine subunits of which a minimum of two are gamma subunits. PMID- 10480949 TI - Interactions of bacteriophage T4-coded primase (gp61) with the T4 replication helicase (gp41) and DNA in primosome formation. AB - One primase (gp61) and six helicase (gp41) subunits interact to form the bacteriophage T4-coded primosome at the DNA replication fork. In order to map some of the detailed interactions of the primase within the primosome, we have constructed and characterized variants of the gp61 primase that carry kinase tags at either the N or the C terminus of the polypeptide chain. These tagged gp61 constructs have been probed using several analytical methods. Proteolytic digestion and protein kinase protection experiments show that specific interactions with single-stranded DNA and the T4 helicase hexamer significantly protect both the N- and the C-terminal regions of the T4 primase polypeptide chain against modification by these procedures and that this protection becomes more pronounced when the primase is assembled within the complete ternary primosome complex. Additional discrete sites of both protection and apparent hypersensitivity along the gp61 polypeptide chain have also been mapped by proteolytic footprinting reactions for the binary helicase-primase complex and in the three component primosome. These studies provide a detailed map of a number of gp61 contact positions within the primosome and reveal interactions that may be important in the structure and function of this central component of the T4 DNA replication complex. PMID- 10480950 TI - E2/E3-mediated assembly of lysine 29-linked polyubiquitin chains. AB - Polyubiquitin (Ub) chains linked through Lys-48-Gly-76 isopeptide bonds represent the principal signal by which substrates of the Ub-dependent protein degradation pathway are targeted to the 26 S proteasome, but the mechanism(s) whereby these chains are assembled on substrate proteins is poorly understood. Nor have assembly mechanisms or definitive functions been assigned to polyubiquitin chains linked through several other lysine residues of ubiquitin. We show that rabbit reticulocyte lysate harbors enzymatic components that catalyze the assembly of unanchored Lys-29-linked polyubiquitin chains. This reaction can be reconstituted using the ubiquitin-conjugating enzyme (E2) known as UbcH5A, a 120-kDa protein(s) that behaves as a ubiquitin-protein ligase (E3), and ubiquitin-activating enzyme (E1). The same partially purified E3 preparation also catalyzes the assembly of unanchored chains linked through Lys-48. Kinetic studies revealed a K(m) of approximately 9 microM for the acceptor ubiquitin in the synthesis of diubiquitin; this value is similar to the concentration of free ubiquitin in most cells. Similar kinetic behavior was observed for conjugation to Lys-48 versus Lys 29 and for conjugation to tetraubiquitin versus monoubiquitin. The properties of these enzymes suggest that there may be distinct pathways for ubiquitin-ubiquitin ligation versus substrate-ubiquitin ligation in vivo. PMID- 10480951 TI - Sulindac inhibits activation of the NF-kappaB pathway. AB - Sulindac is a non-steroidal anti-inflammatory agent that is related both structurally and pharmacologically to indomethacin. In addition to its anti inflammatory properties, sulindac has been demonstrated to have a role in the prevention of colon cancer. Both its growth inhibitory and anti-inflammatory properties are due at least in part to its ability to decrease prostaglandin synthesis by inhibiting the activity of cyclooxygenases. Recently, we demonstrated that both aspirin and sodium salicylate, but not indomethacin, inhibited the activity of an IkappaB kinase beta (IKKbeta) that is required to activate the nuclear factor-kappaB (NF-kappaB) pathway. In this study, we show that sulindac and its metabolites sulindac sulfide and sulindac sulfone can also inhibit the NF-kappaB pathway in both colon cancer and other cell lines. Similar to our previous results with aspirin, this inhibition is due to sulindac-mediated decreases in IKKbeta kinase activity. Concentrations of sulindac that inhibit IKKbeta activity also reduce the proliferation of colon cancer cells. These results suggest that the growth inhibitory and anti-inflammatory properties of sulindac may be regulated in part by inhibition of kinases that regulate the NF kappaB pathway. PMID- 10480952 TI - Direct transport of newly synthesized HLA-DR from the trans-Golgi network to major histocompatibility complex class II containing compartments (MIICS) demonstrated using a novel tyrosine-sulfated chimera. AB - Binding of antigenic peptides to major histocompatibility complex (MHC) class II glycoproteins occurs in specialized endocytic compartments of antigen-presenting cells, which in man are termed MIICs. Newly synthesized MHC class II molecules are transported from the trans-Golgi network to MIICs, but previous studies of this important step in antigen processing have failed to conclusively determine whether most immature MHC class II complexes are transported directly to the processing compartments or are first transiently exposed at the cell surface. To attempt to resolve this question, I constructed a chimeric HLA-DRalpha chain containing two optimal tyrosine sulfation motifs. When expressed in a human B lymphoblastoid cell line lacking functional DRalpha chains, the chimera was correctly incorporated into complexes containing endogenous beta and invariant chains, transported to the trans-Golgi network, and efficiently sulfated. Pulse chase experiments showed that the sulfated complexes were rapidly transported to processing compartments with kinetics consistent with direct transport from the trans-Golgi network. The rate of maturation was not significantly altered in cells expressing a temperature-sensitive mutant of dynamin under conditions where the endocytosis of transferrin was inhibited by 95%, confirming that endocytosis was not required for delivery to MIICs. Maturation of MHC class II-containing complexes was inhibited by aluminum fluoride and brefeldin A, indicating the involvement of heterotrimeric G-proteins and ADP-ribosylation factor in the transport event(s). The procedure described provides a unique mechanism to study critical events in antigen processing and presentation. PMID- 10480953 TI - gamma-aminobutyric acid type B receptor splice variant proteins GBR1a and GBR1b are both associated with GBR2 in situ and display differential regional and subcellular distribution. AB - The subunit architecture of gamma-aminobutyric acid, type B (GABA(B)), receptors in situ is largely unknown. The GABA(B) receptor variants, characterized by the constituents GBR1a and GBR1b, were therefore analyzed with regard to their subunit composition as well as their regional and subcellular distribution in situ. The analysis was based on the use of antisera recognizing selectively GBR1a, GBR1b, and GBR2. Following their solubilization, GBR1a and GBR1b were both found by immunoprecipitation to occur as heterodimers associated with GBR2. Furthermore, monomers of GBR1a, GBR1b, or GBR2 were not detectable, suggesting that practically all GABA(B) receptors are heterodimers in situ. Finally, there was no evidence for an association of GBR1a with GBR1b indicating that these two constituents represent two different receptor populations. A size determination of solubilized GABA(B) receptors by sucrose density centrifugation revealed two distinct peaks of which one corresponded to dimeric receptors, and the higher molecular weight peak pointed to the presence of yet unknown receptor-associated proteins. The distribution and relative abundance of GBR2 immunoreactivity corresponded in all brain regions to that of the sum of GBR1a and GBR1b, supporting the view that most if not all GBR1 proteins are associated with GBR2. However, GBR1a was present preferentially at postsynaptic densities, whereas GBR1b may be mainly attributed to presynaptic or extrasynaptic sites. Thus, GBR1a and GBR1b are both associated with GBR2 to form heterodimers at mainly different subcellular locations where they are expected to subserve different functions. PMID- 10480955 TI - A new semisynthesis of paclitaxel from baccatin III PMID- 10480954 TI - Generation of catalytically active granzyme K from Escherichia coli inclusion bodies and identification of efficient granzyme K inhibitors in human plasma. AB - Granzymes are granule-stored lymphocyte serine proteases that are implicated in T and natural killer cell-mediated cytotoxic defense reactions after target cell recognition. A fifth human granzyme (granzyme 3, lymphocyte tryptase-2), renamed as granzyme K (gene name GZMK), has recently been cloned from lymphocyte tissue. For its further characterization we successfully generated catalytically active enzyme in milligram quantities per liter of Escherichia coli culture. The natural proform of granzyme K with the amino-terminal propeptide Met-Glu was expressed as inclusion bodies and converted to its active enzyme by cathepsin C after refolding of precursor molecules. Recombinant granzyme K cleaves synthetic thiobenzyl ester substrates after Lys and Arg with k(cat)/K(m) values of 3.7 x 10(4) and 4.4 x 10(4) M(-1) s(-1), respectively. Granzyme K activity was shown to be inhibited by the synthetic compounds Phe-Pro-Arg-chloromethyl ketone, phenylmethylsulfonyl fluoride, PefablocSC, and benzamidine, by the Kunitz-type inhibitor aprotinin and by human blood plasma. The plasma-derived inter-alpha trypsin inhibitor complex, its bikunin subunit, and the second carboxyl-terminal Kunitz-type domain of bikunin were identified as genuine physiologic inhibitors with K(i) values of 64, 50, and 22 nM, respectively. Inter-alpha-trypsin inhibitor and free bikunin have the potential to neutralize extracellular granzyme K activity after T cell degranulation and may thus control unspecific damage of bystander cells at sites of inflammatory reactions. PMID- 10480956 TI - New 3-methoxyflavones, an iridoid lactone, and a flavonol from duroia hirsuta PMID- 10480957 TI - Recombinant human manganese superoxide dismutase attenuates early but not delayed skeletal muscle dysfunction following reperfusion injury. AB - OBJECTIVES: to assess the efficacy of recombinant human manganese superoxide dismutase (rhMnSOD) in prevention of early and late skeletal muscle ischaemia reperfusion injury mediated by superoxide (O2-). Design : randomised controlled trial. MATERIALS: seventy-two Sprague-Dawley rats (250-350 g) randomised to receive either 7.5 mg/kg of rhMnSOD or saline. Four hours of ischaemia was induced in the cremaster muscle by dissecting free and clamping its vascular supply. Cremaster muscle contractile function was assessed following 90 minutes, 24, 48 hours and one week of reperfusion. Electrophysiological muscle function was assessed using electrical field stimulation in an organ bath system. RESULTS: muscle function in the untreated groups following ischaemia reperfusion was significantly reduced at 90 minutes, 24, 48 hours and one week of reperfusion (p <0.05). rhMnSOD significantly protected and maintained normal muscle function at 24 and 48 hours (p <0.001). However at one week of reperfusion there was a reduction in function of the treated muscle, such that there was no significant difference between treated and untreated muscle at this point in time. CONCLUSIONS: these data demonstrate that skeletal muscle dysfunction after ischaemia reperfusion injury is attenuated at 24 and 48 hrs of reperfusion by the superoxide scavenger rhMnSOD. This protective effect is not maintained after seven days of reperfusion. PMID- 10480958 TI - Pressure sore prevention--past present and future. PMID- 10480959 TI - Setting up wound dressing guidelines: avoiding the pitfalls. AB - Wound dressings guidelines promote rational prescribing. The development of such guidelines is described, including how to get started, the collection of data, format, contents and avoiding the pitfalls. After implementation, the guidelines require on-going monitoring, review, updating and audit. PMID- 10480960 TI - A study of the performance of a pressure reducing foam mattress after three years of use. AB - This paper details the follow up clinical evaluation of a pressure reducing foam mattress (PRFM) three years after a randomised controlled trial. The PRFMs involved in the 1993 randomised controlled trial were used constantly for three years and recalled in 1996. In the original trial the control group had consisted of 18 standard NHS mattresses. In 1996 none of these mattresses were still in use which made it impossible to replicate the 1993 randomised controlled trial. Instead a clinical evaluation using the 1993 methods was carried out using the PRFMs only. Recruitment to the evaluation proved difficult because changes in clinical practice between the two studies had resulted in shorter lengths of stay in hospital for the target population. Comparison of the two outcome measures namely pressure sore incidence and the subjects' perception of comfort showed no statistically significant change between 1993 and 1996. With the exception of two damaged mattress covers all of the original foam components and covers remained in working order in 1996. During the time between the two studies, a pressure sore prevention programme was implemented within the research area. This programme has resulted in a year by year drop in the Directorate incidence rate. It is therefore not possible to attribute the low pressure sore incidence rate to the PRFMs alone. The results obtained suggest that the PRFMs appear to be providing a similar level of performance after three years of use. PMID- 10480961 TI - Rehabilitation of amputees: clinical management in the brave New World. PMID- 10480962 TI - Morphological characteristics of the dermal papillae in the development of pressure sores. AB - Mechanisms of skin break down in the development of human pressure sores are still unclear. This study was undertaken to clarify the morphological characteristics of the dermal papillae in the skin associated with pressure sores. Skin tissues were excised from the sacrum of a Japanese subject post mortem, where a superficial pressure sore had developed. Light microscopic and transmission and scanning electron microscopic examinations were performed. It was found that the atrophic, irregular contour and alignment of the dermal papillae were characteristic of the boundary area between healthy and damaged areas. In addition, a relatively dense network of collagen fibres in the papillary layer of the boundary area was observed when compared with the healthy area. These findings suggest that the morphological changes of the papillae observed in the boundary area affect microcirculation, impairing tissue viability by inhibiting nutritive blood supply and by accumulating metabolic byproducts which predispose to tissue damage. PMID- 10480963 TI - Tissue viability and the pharmacist. PMID- 10480964 TI - Tissue viability. PMID- 10480965 TI - Hyperaemia. AB - Hyperaemia is the process by which the body adjusts blood flow to meet the metabolic needs of its different tissues in health and disease. Meticulous control of the microcirculation--the arterioles, capillaries and venules--is essential to life. Reactive hyperaemia, the local vasodilatation which occurs in response to oxygen debt and accumulation of metabolic waste products due to interruption of blood flow; active hyperaemia, the increased blood flow in an organ during a period of activity; and the hyperaemic response to infection and trauma are vitally important. The microcirculation is controlled partly by sympathetic vasoconstrictor impulses from the brain and partly by vasoactive substances secreted locally by the endothelial cells. The most important of the latter is nitric oxide which facilitates flow by causing relaxation of vascular smooth muscle. Neural and endothelial control of blood flow are impaired by illness. Neurological disease and vascular disease which affect the microcirculation, predispose patients to develop ischaemic organ damage, including pressure sores, during periods of intercurrent illness. Severe sepsis or trauma may cause irreversible microcirculatory dysfunction resulting in multi organ failure and death. PMID- 10480966 TI - The past politics of pressure sores. PMID- 10480968 TI - Pressure measurements in the management of tissue viability. PMID- 10480967 TI - Development and validation of the Burton Score: a tool for nutritional assessment. AB - This paper describes the development and validation of the Burton Score, a nutritional assessment tool based on the Waterlow score, with the rationale that since nurses already collect data for one score, it would only lead to unnecessary duplication of effort if a totally different scoring scheme were to be used for nutritional assessment. Initial cut offs were determined by a pilot study of 26 patients on an elderly care ward and validated by comparing the nutritional status of 263 patients estimated by the Burton score with a dietitian's assessment of nutrition. The validation study showed that although there was significant correlation between the Burton and Waterlow scores the Burton score correlated more closely with the dietitian's assessments. The King's Fund report of 1992 stated that all patients should have assessment of nutritional status on admission to hospital: we believe the Burton score could provide a simple tool to achieve this goal. PMID- 10480969 TI - Professional roles--the nurse educator: a personal view. PMID- 10480970 TI - The Chronic Wound Support Service. PMID- 10480971 TI - Review of duplex and colour Doppler imaging of lower-limb arteries and veins. AB - Ultrasonic imaging provides a non-invasive assessment of the arterial and venous circulation in the lower limb and is accepted as a valuable diagnostic technique. Grey-scale images identify plaque and thrombus, duplex assessment provides a measurement of blood velocity through a vessel, and colour Doppler imaging enables the rapid localization of arterial stenoses and occlusions and the identification of incompetent veins. This article outlines the principles of the different techniques and presents normal images. Procedures for investigating arterial stenoses, superficial venous incompetence and deep venous thrombosis are described, abnormal images presented and the limitations discussed. It is hoped to provide an insight into the strengths and limitations of ultrasonic vascular investigations for those involved in tissue viability and ulcer management. PMID- 10480972 TI - The effect of surveillance on surgical wound infection rates. AB - The clean wound infection rate is the most valuable reflection of surgical care in any hospital. Wound infection rates are seen as indicators of quality in terms of negative outcome measures. A nurse was appointed by Hairmyres and Stonehouse Hospitals NHS Trust for the purpose of surveillance of the clean surgical wound infection rate. The surveillance is based on a previous study in the USA showing that feedback of infection rates to surgeons can lead to a reduction in these rates. The specifications for surveillance were that all clean surgery would be followed up for 30 days post-operatively and that wounds with implants, i.e. vascular grafts, would be followed up for 1 year post-operatively to produce infection rates for the surgeons, wards, theatres and the infection control team. Data collection commenced in October 1995 and so far 1851 clean surgical cases have been followed up for 30 days post-operatively. All patients were monitored until discharge, then at 30 days post-operatively, and the wound scored for signs of infection. Day cases were seen within week 1 and at day 30. General practitioner and district nurse liaison enabled detection of problems at other times. The patients were given a telephone number they could use to report problems after surgery. The results show that this type of surveillance is an effective way of collecting accurate data on wound infection rates. It has enabled the early identification of problem areas in practice. The audit has also been used to facilitate the adoption of evidence-based practice, through recommendations for clean surgery, to reduce the extrinsic risk factors for wound infection. As a result of the surveillance, there has been a significant reduction (P < 0.05) in the clean wound infection rate. PMID- 10480973 TI - The diagnosis of early pressure sores: report of the pilot study. AB - Thirty-four vascular and general surgical patients were recruited to a pilot study exploring skin blood flow using laser Doppler imaging and clinical assessment of skin erythema in relation to pressure sore development. Brief details of the results, sample size calculations and main study methodology are described in this mid-term report of the Tissue Viability Society Research Fellowship (1998-2000). PMID- 10480974 TI - Laser therapy for the treatment of venous leg ulcers. PMID- 10480975 TI - Test your knowledge. Tinea corporis. PMID- 10480976 TI - An essential element of implementing evidence-based practice (EBP) is a definition of what evidence is and how different types of evidence compare to one another. PMID- 10480977 TI - The relationship between diabetic amputees and vascular reviews. AB - This paper reviews the data in the literature pertaining to risk factors for amputation in the diabetic population, and describes a retrospective study to assess the vascular and other assessments carried out on a sample of 50 diabetic patients undergoing minor (toe and foot) and major (below-knee and above-knee) amputations. The study found that 50% of the patients did not have Doppler studies, 52% did not have angiography, and 78% did not have angioplasty. Major amputations were performed in 76% of the sample. This is an unacceptably high figure. Thorough vascular assessments need to be performed in an attempt to save lower limbs from potential amputation. PMID- 10480978 TI - CE marking--what does it really mean? AB - Compliance with the Medical Devices Directive (93/42/EEC) became compulsory for all medical devices placed on the European market in June 1998. Under the directive, manufacturers are required to fulfil a number of requirements leading to the placing of the CE mark on their devices. However, many questions still remain. Not all manufacturers are aware of their responsibilities and not all purchasers are aware of the true meaning of the CE mark. It is vital to understand the purpose of the regulations, the definition of 'medical device', the importance of complying with the manufacturer's intended purpose and instructions, the role of quality systems and clinical investigations in the compliance process, and the decisions which may be made by the manufacturers. It is also important to realise that whilst the directive, if correctly applied and enforced, brings benefits to the medical industry and patients, it is not a substitute for professional judgement in the purchasing process. PMID- 10480979 TI - From two weeks to twenty-two: one patient's experience of minor surgery. PMID- 10480980 TI - Zinc and chronic leg ulcers: a systematic review of oral zinc in the treatment of chronic leg ulcers. PMID- 10480981 TI - From the archives. Dressings and strapping for wounds. PMID- 10480982 TI - Adrenomedullin in nonmammalian vertebrate pancreas: an immunocytochemical study. AB - Adrenomedullin (AM) immunoreactive cells have been identified, by immunocytochemical methods, in the endocrine pancreas of seven nonmammalian vertebrate species, belonging to the cartilaginous and bony fish, amphibian, reptilian, and bird classes. The frequency and distribution of the pancreatic AM cells vary among the different animals. In most species, these cells are found scattered mainly among the exocrine component, with a few present in the islet like structures. The distribution of AM cells in both fish species and Xenopus shows an inverse pattern, since almost every AM cell is located in the islets. In addition, the colocalization of AM with other classical pancreatic peptide immunoreactivities has been analyzed. In numerous cells, AM immunoreactivity did not colocalize with the other hormones, suggesting that AM-producing cells might constitute a new endocrine cell type in the pancreas of many species. Nevertheless, in other cells a species-specific pattern of colocalizations with insulin, somatostatin, glucagon, and pancreatic polypeptide was found, indicating that complex interactions among all these hormones may occur. In conclusion, AM represents a new regulatory peptide of the endocrine nonmammalian vertebrate pancreas, which is possibly involved in the modulation of insulin secretion and other pancreatic functions. PMID- 10480983 TI - Monitoring of progesterone in captive female false killer whales, Pseudorca crassidens. AB - The present study describes progesterone profiles to enhance understanding of general reproductive patterns in three female captive false killer whales and analyzes potential relationships in progesterone concentrations between plasma, salivary, and ocular secretions. Plasma progesterone concentrations reflected ovarian activity for most of the year, with increased concentrations in the spring and summer, indicating that the two adult female false killer whales were spontaneous ovulators and seasonally polyestrus. Elevated progesterone concentrations were determined at intervals, for up to 10 consecutive months, in one female. There were also varying periods of no apparent ovarian activity from 3 to 10 consecutive months. Correlation coefficients between progesterone concentrations in plasma, salivary, and ocular secretions ranged between -0.23 and 0.16. It is concluded that blood collection should not be replaced by salivary or ocular secretion collection for the measurement of progesterone in the false killer whale. PMID- 10480984 TI - Molecular cloning and pharmacological characterization of a somatostatin receptor subtype in the gymnotiform fish Apteronotus albifrons. AB - The actions of the various forms of somatostatin (SRIF), including those of the tetradecapeptide SRIF(14), are mediated by specific receptors. In mammals, five subtypes of SRIF receptors, termed sst(1-5), have been cloned. Using a combination of reverse transcriptase-polymerase chain reaction and genomic library screening in the gymnotiform fish Apteronotus albifrons, a gene encoding the first-known nonmammalian SRIF receptor has been isolated. The deduced amino acid sequence displays 59% identity with the human sst(3) receptor protein; hence, the gene is termed "Apteronotus sst(3)." The predicted protein consists of 494 amino acid residues exhibiting a putative seven-transmembrane domain topology typical of G protein-coupled receptors. A signal corresponding to the Apteronotus sst(3) receptor was detected in brain after amplification of poly(A)(+)-RNA by reverse transcriptase-polymerase chain reaction, but not by Northern blot analysis or in situ hybridization, suggesting a low level of expression. Membranes prepared from CCL39 cells stably expressing the Apteronotus sst(3) receptor gene bound [(125)I][Leu(8),d-Trp(22), (125) I-Tyr(25)]SRIF(28) with high affinity and in a saturable manner (B(max) = 4470 fmol/mg protein; pK(D) = 10.5). SRIF(14) and various synthetic SRIF receptor agonists produced a dose-dependent inhibition of radioligand binding, with the following rank order of potency: SRIF(14) approximately SRIF(28) > BIM 23052 > octreotide > BIM 23056. Under low stringency conditions, an Apteronotus sst(3) probe hybridized to multiple DNA fragments in HindIII or EcoRI digests of A. albifrons DNA, indicating that the Apteronotus sst(3) receptor is a member of a larger family of Apteronotus SRIF receptors. PMID- 10480985 TI - Effects of embryonic treatment with fadrozole on phenotype of gonads, syrinx, and neural song system in zebra finches. AB - Previous studies have found that treatment of zebra finch embryos with an aromatase inhibitor on Day 5 or 8 of incubation caused partial sex reversal of gonadal phenotype in females. These females possessed both testicular and ovarian tissue, and the development of the neural circuit for song remained feminine. The present study attempted more complete gonadal reversal by treating zebra finch embryos earlier, on Day 3 of incubation, with Fadrozole (CGS 16949A), an aromatase inhibitor, or with saline. We examined the phenotype of the syrinx (androgen-dependent vocal organ), the gonads, and the telencephallic neural song system in 100-day-old birds. Treated females typically possessed a left ovotestis and a right testis, and significantly larger syringes than control females. The histology and steroid synthetic enzyme activity of the testicular tissue in treated females were quite masculine and similar to that of control males. At the time of sacrifice, the plasma concentrations of testosterone and estradiol for fadrozole-treated females did not differ from those of control females, but dihydrotestosterone was lower in treated females. Despite the large amount of functional testicular tissue and a masculine syrinx, the volumes and soma sizes of song system nuclei (HVC, RA) in treated females remained feminine. These results suggest that testicular secretions masculinize the syrinx, but are not sufficient to masculinize the song system in zebra finches. PMID- 10480986 TI - The chicken leptin gene: has it been cloned? AB - The DNA sequence of a chicken leptin gene that shares 95% nucleotide similarity with the mouse leptin sequence has been recently reported (Taouis et al., 1998, Gene 208, 239-242). Experiments have been performed independently in two laboratories to try to confirm this finding. Fourteen PCR primers based on the mouse leptin sequence were designed to amplify the avian leptin gene. Four of the primers were identical to the mouse and published chicken leptin sequences. PCR amplification was carried out on genomic DNA and reverse-transcribed mRNA from the fat, liver, and pancreas of several chicken strains and from the domestic turkey, goose, and Japanese quail. No PCR products sharing close similarity to the mouse leptin sequence were generated from any avian templates. Amplification of mouse leptin sequence was consistently obtained when control mouse templates were used. Northern hybridization using a mouse leptin probe failed to produce a signal with poly(A)+ RNA from chicken fat and liver and from the fat and liver of force-fed geese but a strong signal was obtained from control mouse fat total RNA. Southern hybridization under low stringency washing conditions revealed hybridization of a mouse leptin probe to chicken genomic DNA. Under higher stringency washing conditions, the chicken signal disappeared, while those from control mouse and sheep genomic DNA remained. This suggests that the putative chicken leptin sequence shares less than the 83% nucleotide sequence identity between the mouse and sheep genes. It is concluded that a chicken leptin gene sequence with close sequence similarity to mouse leptin is not present in the chicken genome. Furthermore, mRNA sharing high sequence identity with mouse leptin is not present in the fat or liver of the domestic chicken, turkey, goose, or Japanese quail. PMID- 10480987 TI - Influence of corticosterone on FSH-induced ovarian recrudescence in the lizard Mabuya carinata. AB - Administration of bovine FSH (10 IU/lizard/alternate day for 30 days) in the postbreeding quiescent phase of the ovarian cycle caused a significant increase in the mean number of oogonia and oocytes, the relative weight of the oviduct, and the liver and serum estradiol levels compared to those of controls. In addition, the FSH-treated lizards showed a vitellogenic growth of follicles and development through to preovulatory follicles. However, the administration of corticosterone simultaneously with FSH (10 IU FSH + 40 microgram corticosterone/lizard/alternate day for 30 days) did not result in these changes and the ovaries resembled those of controls. The results indicate the absence of ovarian refractoriness to gonadotropic stimulation during the quiescent phase of the reproductive cycle and inhibition of FSH-induced ovarian recrudescence by corticosterone. It is suggested that corticosterone treatment reduces FSH-induced steroidogenic activity of the ovary, leads to impairment in vitellogenin secretion by the liver, and results as well in the failure of vitellogenic follicular growth in Mabuya carinata. PMID- 10480988 TI - Androgenic gland hormone is a sex-reversing factor but cannot be a sex determining factor in the female crustacean isopods Armadillidium vulgare. AB - Sex reversal of female isopods, Armadillidium vulgare, has been induced by implantation of the androgenic gland (AG) into individuals after the initiation of morphological sex differentiation. The focus of the present study is to examine whether female gonads are reversed by the androgenic gland hormone (AGH) during the sexually undifferentiated period through postembryonic development in A. vulgare. Instead of injections of AGH, three AGs were implanted into each genetic female at various developmental stages to induce sex reversal. Before implantation fresh AGs were treated with ethanol to stop AGH synthesis, but then still contained AGH. These AGs have been referred to as ethanol-treated AGs (t AGs). Development of a testis was used as an indicator of gonadal sex reversal. The gonads of genetic females were transformed into testes by implantations of t AGs during the sex differentiation period. However, when genetic females received implants at sexually undifferentiated stages, development of their gonads was not reversed in the male direction. These results suggest that after the onset of gonadal sex differentiation, AGH is a sex-reversing factor that can turn a female gonad into a male gonad. AGH cannot be a sex-determining factor in female A. vulgare, as undifferentiated gonads of genetic females are not sex reversed by the hormone. PMID- 10480989 TI - Effect of T(3) treatment and food ration on hepatic deiodination and conjugation of thyroid hormones in rainbow trout, Oncorhynchus mykiss. AB - We studied the 7-day effects of 3,5,3'-triiodothyronine (T(3)) hyperthyroidism (induced by 12 ppm T(3) in food) and food ration (0, 0.5, or 2% body weight/day) on in vitro hepatic glucuronidation, sulfation, and deiodination of thyroxine (T(4)), T(3), and 3,3', 5'-triiodothyronine (rT(3)). T(3) treatment doubled plasma T(3) with no change in plasma T(4), depressed hepatic low-K(m) (1 nM) outer-ring deiodination (ORD) of T(4), induced low-K(m) (1 nM) inner-ring deiodination (IRD) of both T(4) and T(3) but did not alter high-K(m) (1 microM) rT(3)ORD, glucuronidation, or sulfation of T(4), T(3), or rT(3). Plasma T(4) levels were greater for 0 and 2% rations than for a 0.5% ration. Fasting decreased low-K(m) T(4)ORD activity and increased high-K(m) rT(3)ORD activity but did not alter T(4)IRD or T(3)IRD activities. T(4), T(3), and rT(3) glucuronidation were greater for 0 and 0.5% rations than for a 2% ration. T(3) glucuronidation was greater for a 0.5% ration than for a 0% ration. T(3) and rT(3) sulfation were greater for a 2% ration than for a 0 or a 0.5% ration; ration did not change T(4) sulfation. We conclude that (i) modest experimental T(3) hyperthyroidism induces T(3) autoregulation by adjusting hepatic low-K(m) ORD and IRD activities but not high-K(m) rT(3)ORD or conjugation activities; (ii) in contrast, ration level changes both deiodination and conjugation pathways, suggesting that the response to ration does not solely reflect altered T(3) production; (iii) deiodination and conjugation appear complementary in regulating thyroidal status in response to ration; and (iv) high-K(m) rT(3)ORD in trout differs from rat type I deiodination in that it does not respond to T(3) hyperthyroidism and it increases, rather than decreases, its activity during fasting. PMID- 10480990 TI - Deiodination and deconjugation of thyroid hormone conjugates and type I deiodination in liver of rainbow trout, Oncorhynchus mykiss. AB - We studied the hepatic in vitro deconjugation and deiodination of glucuronide (G) and sulfate (S) conjugates of the thyroid hormones (TH) thyroxine (T(4)), 3,5,3' triiodothyronine (T(3)), and 3,3', 5'-triiodothyronine (rT(3)) in trout. These conversions have not been studied in nonmammals. Deconjugation of T(4)G, T(3)G, rT(3)G, or rT(3)S was negligible in all subcellular fractions. Some T(4)S desulfation occurred but T(3)S was desulfated to the greatest extent by freshly isolated hepatocytes and by the mitochondrial/lysosomal and microsomal fractions. Deiodination of T(4)G, T(3)G, rT(3)G, T(4)S, T(3)S, and rT(3)S (1 or 1000 nM) was negligible in control trout and in trout treated with T(3) to induce inner-ring deiodination (IRD) but simultaneously tested rat microsomes rapidly deiodinated T(4)S, T(3)S, and rT(3)S. Furthermore, T(4)S, T(3)S, and rT(3)S (1-100 nM) were less effective than their unsulfated forms in competitively inhibiting trout hepatic outer-ring deiodination (ORD) of T(4) (0.8 nM), and rT(3)ORD (100 nM) was not competitively inhibited by T(4)S, T(3)S, or rT(3)S (100 nM) or by T(4) or T(3) (1 microM). Thus, there is no evidence in trout liver for THS deiodination, which is a key property of rat type I deiodination. We therefore studied other properties of trout hepatic high-K(m) deiodination, which has been considered homologous to rat type I deiodination. We found that it resembled rat type I deiodination in its rT(3)ORD ability, its optimum pH (7.0), and its requirement for dithiothreitol (DTT). However, it differed from rat type I deiodination not only in its negligible deiodination of T(4) and THS but also in its low DTT optimum (2.5 mM), its low apparent K(m) for rT(3) (200 nM), its lack of IRD ability, its extremely weak propylthiouracil inhibition (IC(50), 1 mM), its weaker inhibition by iodoacetate (IC(50), 10 microM) and aurothioglucose (IC(50), <3 microM), its activation by fasting, and its unresponsiveness to T(3) hyperthyroidism. We conclude that most conjugated TH are neither deconjugated nor deiodinated by trout liver and are therefore eliminated in bile. However, T(3)S can be desulfated. Substrate preference and other properties suggest that trout hepatic high-K(m) ORD shares some properties with rat type I deiodination but differs functionally in several other respects and may contribute negligibly to hepatic T(3) production in trout. PMID- 10480991 TI - Free and protein-bound insulin-like growth factor-I (IGF-I) and IGF-binding proteins in plasma of coho salmon, Oncorhynchus kisutch. AB - Total and free insulin-like growth factor-I (IGF-I) levels were quantified in plasma from growth hormone (GH)-treated and fasted coho salmon. Total IGF-I was measured by radioimmunoassay after acid-ethanol extraction and free IGF-I was separated from protein-bound IGF-I using ultrafiltration by centrifugation. Total and free IGF-I increased in plasma after GH treatment and decreased after fasting. The level of free IGF-I, however, was maintained at approximately 0.3% in both experiments. Unsaturated binding activity in plasma for IGF-I was assessed by incubation with (125)I-recombinant salmon IGF-I ((125)I-sIGF-I). Although there was no difference in binding activity between GH-treated and control fish, fasted fish showed higher binding activity than did fed fish, suggesting induction of unsaturated binding protein by fasting. IGF-binding protein (IGFBP) bands were observed in plasma of coho salmon by Western ligand blotting using (125)I-sIGF-I. A low-molecular-weight (22 kDa) band was clear in fasted fish but not detectable in fed fish. The IGFBP band, which has molecular weight similar to that of human IGFBP-3 (41 kDa), was more intense in GH-treated fish than in controls. The molecular distribution of IGF-I in plasma was examined by gel filtration under neutral conditions. Most IGF-I was eluted around 40 kDa. This result suggests that the major form of bound IGF-I in the circulation of coho salmon may be in a 40-kDa binary complex rather than in a 150-kDa ternary complex, as in mammals. PMID- 10480992 TI - Isolation and amino acid sequence of a peptide with vitellogenesis inhibiting activity from the terrestrial isopod Armadillidium vulgare (Crustacea). AB - The complete amino acid sequence of a neuropeptide was established using gas phase microsequencing, mass spectrometry, and reverse transcription-polymerase chain reaction. This peptide, stored in the sinus gland of the terrestrial isopod Armadillidium vulgare, inhibited vitellogenin synthesis by the fat tissue and inhibited the onset of secondary vitellogenesis when tested in homologous bioassays. This peptide, named Arv-VIH, has 83 amino acid residues and a molecular mass of 9485 Da. Relationships with other related peptides are presented. PMID- 10480993 TI - Molecular cloning of lungfish proopiomelanocortin cDNA. AB - To investigate the evolution of proopiomelanocortin (POMC) from fish to tetrapods, nucleotide sequence of POMC cDNA from a lobe-finned fish, the African lungfish, was determined. POMC cDNA was prepared from lungfish pituitary glands. The POMC cDNA is composed of 1114 bp, excluding a poly-A tail, and encodes 255 amino acids (aa) including a signal peptide of 25 aa. The lungfish POMC contains the segment corresponding to gamma-melanotropin (MSH), corticotropin, alpha-MSH, beta-MSH, and beta-endorphin at positions (50-61), (108-146), (108-120), (178 194), and (197-230), respectively. The lungfish POMC shows greater sequence identity on average with amphibian (62%), ancient ray-finned fishes including acipenseriformes and semionotiformes (62%), and mammalian POMC (52%) than with teleostean (49%), elasmobranch (46%), and agnathan POMC (31%). Thus, the overall structural feature of lungfish POMC is close to the tetrapod POMCs which contain gamma-MSH and the ancient ray-finned fishes POMCs containing gamma-MSH-like sequence. However, amino acid sequence of lungfish beta-endorphin exhibits properties which are specifically observed in the ray-finned fishes and the elasmobranchs. PMID- 10480994 TI - Characterization of the cloned atlantic cod neuropeptide Y-Yb receptor: peptide binding requirements distinct from known mammalian Y receptors. AB - Five members of the neuropeptide Y (NPY) receptor family have been cloned in mammals. The recently cloned NPY receptor in the Atlantic cod seems to be distinct from the mammalian subtypes as it has only 50% identity to Y1, Y4, and y6 and only 30% to Y2 and Y5. In most of the other families of G-protein-coupled receptors, species homologues have 65-90% identity between fishes and mammals. The functional expression and detailed pharmacological characterization of this cod NPY receptor, designated Yb, is reported. Membranes of cells transiently transfected with cod Yb showed saturable [(125)I]PYY binding with a K(d) of 45 pM. The pharmacological profile is similar to those of both the zebrafish Yb and Yc receptors and distinct from those of the mammalian NPY receptors. In competition experiments the cod Yb receptor had the following rank order of potencies: porcine PYY = porcine NPY = p[Leu(31), Pro(34)]NPY > zebrafish PYY > zebrafish NPY >> NPY2-36 = NPY3-36 > NPY18-36 > bovine PP = [D-Trp(32)]NPY > BIBP3226. This is in sharp contrast to the high selectivity of BIBP3226 for the Y1 receptor from all mammalian species. Together with the low amino acid identity of cod Yb with the mammalian Y1, Y4, and y6 receptors, this is further support for the notion that fish Yb constitutes a distinct NPY receptor subtype. PMID- 10480995 TI - Distribution of progesterone receptor in female mouse tissues. AB - Two novel antibodies against the mammalian progesterone receptor (PR) were raised and characterized to study the distribution of PR and the effect of estrogen on PR expression in various female murine tissues by immunohistochemistry. There were estrogen-independent constitutive PR expressions in the smooth muscle cells of uterus, uterine blood vessels, urinary bladder, duodenum, and jejunum of ovariectomized mice. Uterine stromal cells, capsular cells of kidney and adrenal gland, and the epithelial cells of submandibular gland expressed PR constitutively. PR expression was detected in some thymic cells and the number of PR-positive thymic cells increased markedly after estrogen treatment. Estrogen induced PR expression in the epithelial cells of uterus, vagina, urethra, and skin and the stromal cells of vagina, urethra, and pancreatic ducts, as well as the smooth muscle cells of some blood vessels. These results suggest cell specific progesterone actions in the urinary tract, skin, and gastrointestinal organs, on the immune functions, and on the regulation of local blood flow. PMID- 10480996 TI - Combined aromatase inhibitor and antiandrogen treatment decreases territorial aggression in a wild songbird during the nonbreeding season. AB - Male song sparrows (Melospiza melodia morphna) defend territories throughout the year in western Washington State. In the nonbreeding season (autumn and winter), aggression and song are robustly expressed but plasma testosterone (T) levels are basal. Also, castration does not decrease nonbreeding territoriality. In this field experiment, we asked whether nonbreeding aggression is independent of T. T can act via androgen receptors or T can be aromatized to 17beta-estradiol (E(2)) and act via estrogen receptors. We treated free-living nonbreeding birds with an aromatase inhibitor (ATD) and an androgen receptor antagonist (flutamide) in combination. We then challenged subjects with a live decoy and playback of tape recorded songs. ATD+flutamide treatment decreased several aggressive behaviors. However, ATD+flutamide treatment did not affect body condition, suggesting that subjects were healthy and that foraging behavior was not reduced. As expected, ATD+flutamide treatment increased plasma T, probably by blocking negative feedback on luteinizing hormone. Surprisingly, ATD+flutamide treatment increased plasma E(2). Most other studies using aromatase inhibitors have not measured plasma E(2). However, it is possible that ATD+flutamide treatment decreased local E(2) levels in the brain but not in plasma. Finally, ATD+flutamide treatment increased plasma corticosterone, perhaps in response to increased plasma T or E(2). To our knowledge, these are the first data to suggest that nonbreeding territoriality is regulated by endogenous steroid hormones. Nongonadal production of sex steroids may support aggression in the nonbreeding season. PMID- 10480997 TI - Regulation of gap junctions and oocyte maturational competence by gonadotropin and insulin-like growth factor-I in ovarian follicles of red seabream. AB - The in vitro effects of human chorionic gonadotropin (hCG) and human insulin-like growth factor-I (IGF-I) on the formation of oocyte-granulosa cell (heterologous) and granulosa cell-granulosa cell (homologous) gap junctions (GJ) were determined during the course of development of oocyte maturational competence (OMC) in ovarian follicles of red seabream. Incompetent, full-grown follicles were incubated in hCG (10 IU/ml), IGF-I (10 nM), or hormone-free medium for 0, 6, and 12 h and examined by electron microscopy. Qualitative analysis indicated that homologous and heterologous GJ were relatively rare in incompetent follicles and remained rare if they were maintained in control medium. However, both types of GJ were readily observed in follicles treated with hCG or IGF-I. For quantitative analysis, we measured changes in the combined numbers of both GJ types, since discrimination between the two was not always possible. The numbers of GJ along 125 micrometer of granulosa cell-zona radiata border were counted in three follicles/treatment. GJ markedly increased with time in the presence of hCG or IGF-I but not in control medium (Duncan's multiple range test; P < 0.05). Functional assays confirmed that the hormonal treatments also induced OMC. Naturally competent follicles sampled at the appropriate stage of the reproductive cycle contained high levels of both types of GJ suggesting that the OMC-associated increase in GJ is physiologically relevant. In view of similar, earlier results with the Atlantic croaker, the present results support the concept that an increase in homologous and heterologous GJ is a general developmental event associated with the acquisition of OMC in teleost ovarian follicles. Also, a novel finding of this study is that IGF-I can regulate heterologous and homologous GJ in intact ovarian follicles. PMID- 10480998 TI - Isolation and characterization of the goldfish thyrotropin beta subunit gene including the 5'-flanking region. AB - The complete gene encoding the beta subunit of thyrotropin (thyroid-stimulating hormone, TSH) was isolated from a goldfish genomic library. The goldfish TSHbeta subunit gene, which is approximately 2.0 kilobase pairs (kb) in length, consisted of three exons and two introns. The first intron was much longer (0.89 kb) than the second intron (0.3 kb) as are TSHbeta genes in mammalian species. On the basis of the location of the first intron, the goldfish TSHbeta gene belongs to the mammalian TSHbeta/FSHbeta gene group, which is distinct from the LHbeta group. Inspection of the 5'-flanking and exon 1 regions of the goldfish TSHbeta gene (1.2 kb) revealed the presence of several putative cis-acting elements, including the negative triiodothyronine (T(3))-responsive element (nTRE), Pit-1 element, and GATA-2 element. Comparison of the goldfish sequence with mammalian TSHbeta promoter sequences showed an identical region, nTRE, in the first exon intron junction region. An in vitro study using dispersed goldfish pituitary cells showed that T(3) treatment (20 ng/ml) suppressed the TSHbeta mRNA level in the cells. These data indicate that (1) the basic structure of TSHbeta genes is highly conserved in vertebrates and that (2) T(3) acts directly on the pituitary and inhibits TSHbeta gene expression in goldfish, probably via the nTRE in the TSHbeta gene. PMID- 10480999 TI - Trends in the vertical distribution of ozone AB - Analyses of satellite, ground-based, and balloon measurements allow updated estimates of trends in the vertical profile of ozone since 1979. The results show overall consistency among several independent measurement systems, particularly for northern hemisphere midlatitudes where most balloon and ground-based measurements are made. Combined trend estimates over these latitudes for the period 1979-96 show statistically significant negative trends at all altitudes between 10 and 45 km, with two local extremes: -7.4 +/- 2. 0% per decade at 40 km and -7.3 +/- 4.6% per decade at 15 km altitude. There is a strong seasonal variation in trends over northern midlatitudes in the altitude range of 10 to 18 km, with the largest ozone loss during winter and spring. The profile trends are in quantitative agreement with independently measured trends in column ozone, the amount of ozone in a column above the surface. The vertical profiles of ozone trends provide a fingerprint for the mechanisms of ozone depletion over the last two decades. PMID- 10481000 TI - Onset of fermi degeneracy in a trapped atomic Gas AB - An evaporative cooling strategy that uses a two-component Fermi gas was employed to cool a magnetically trapped gas of 7 x 10(5) (40)K atoms to 0.5 of the Fermi temperature T(F). In this temperature regime, where the state occupation at the lowest energies has increased from essentially zero at high temperatures to nearly 60 percent, quantum degeneracy was observed as a barrier to evaporative cooling and as a modification of the thermodynamics. Measurements of the momentum distribution and the total energy of the confined Fermi gas directly revealed the quantum statistics. PMID- 10481001 TI - A capacitance standard based on counting electrons AB - A capacitance standard based directly on the definition of capacitance was built. Single-electron tunneling devices were used to place N electrons of charge e onto a cryogenic capacitor C, and the resulting voltage change DeltaV was measured. Repeated measurements of C = Ne/DeltaV with this method have a relative standard deviation of 0.3 x 10(-6). This standard offers a natural basis for capacitance analogous to the Josephson effect for voltage and the quantum Hall effect for resistance. PMID- 10481003 TI - Lower mantle lateral heterogeneity beneath the caribbean Sea AB - Seismic wave reflections from Earth's core recorded at seismic arrays in North America from events in the Caribbean Islands, Venezuela, and the Mid-Atlantic Ridge have observed slownesses more than 64 percent greater than predicted by the IASPEI91 standard Earth model. P waves turning in the lowermost mantle beneath the same region also have anomalous slowness. The slowness anomalies are not accompanied by significant travel time residuals and appear to be caused by lateral inhomogeneities in the velocity structure of the lower mantle. PMID- 10481002 TI - Increased summertime UV radiation in New Zealand in response to ozone loss. AB - Long-term decreases in summertime ozone over Lauder, New Zealand (45 degrees S), are shown to have led to substantial increases in peak ultraviolet (UV) radiation intensities. In the summer of 1998-99, the peak sunburning UV radiation was about 12 percent more than in the first years of the decade. Larger increases were seen for DNA-damaging UV radiation and plant-damaging UV radiation, whereas UV-A (315 to 400 nanometers) radiation, which is insensitive to ozone, showed no increase, in agreement with model calculations. These results provide strong evidence of human-induced increases in UV radiation, in a region where baseline levels of UV radiation were already relatively high. PMID- 10481004 TI - An infrared spectral match between GEMS and interstellar grains. AB - Infrared spectral properties of silicate grains in interplanetary dust particles (IDPs) were compared with those of astronomical silicates. The approximately 10 micrometer silicon-oxygen stretch bands of IDPs containing enstatite (MgSiO3), forsterite (Mg2SiO4), and glass with embedded metal and sulfides (GEMS) exhibit fine structure and bandwidths similar to those of solar system comets and some pre-main sequence Herbig Ae/Be stars. Some GEMS exhibit a broad, featureless silicon-oxygen stretch band similar to those observed in interstellar molecular clouds and young stellar objects. These GEMS provide a spectral match to astronomical "amorphous" silicates, one of the fundamental building blocks from which the solar system is presumed to have formed. PMID- 10481005 TI - Heterostructures of single-walled carbon nanotubes and carbide nanorods AB - A method based on a controlled solid-solid reaction was used to fabricate heterostructures between single-walled carbon nanotubes (SWCNTs) and nanorods or particles of silicon carbide and transition metal carbides. Characterization by high-resolution transmission electron microscopy and electron diffraction indicates that the heterostructures have well-defined crystalline interfaces. The SWCNT/carbide interface, with a nanometer-scale area defined by the cross section of a SWCNT bundle or of a single nanotube, represents the smallest heterojunction that can be achieved using carbon nanotubes, and it can be expected to play an important role in the future fabrication of hybrid nanodevices. PMID- 10481006 TI - Recognition of the codon-anticodon helix by ribosomal RNA. AB - Translational fidelity is established by ribosomal recognition of the codon anticodon interaction within the aminoacyl-transfer RNA (tRNA) site (A site) of the ribosome. Experiments are presented that reveal possible contacts between 16S ribosomal RNA and the codon-anticodon complex. N1 methylation of adenine at position 1492 (A1492) and A1493 interfered with A-site tRNA binding. Mutation of A1492 and A1493 to guanine or cytosine also impaired A-site tRNA binding. The deleterious effects of A1492G or A1493G (or both) mutations were compensated by 2'fluorine substitutions in the mRNA codon. The results suggest that the ribosome recognizes the codon-anticodon complex by adenine contacts to the messenger RNA backbone and provide a mechanism for molecular discrimination of correct versus incorrect codon-anticodon pairs. PMID- 10481007 TI - Activity-induced potentiation of developing neuromuscular synapses. AB - Electrical activity plays a critical role in shaping the structure and function of synaptic connections in the nervous system. In Xenopus nerve-muscle cultures, a brief burst of action potentials in the presynaptic neuron induced a persistent potentiation of neuromuscular synapses that exhibit immature synaptic functions. Induction of potentiation required an elevation of postsynaptic Ca2+ and expression of potentiation appeared to involve an increased probability of transmitter secretion from the presynaptic nerve terminal. Thus, activity dependent persistent synaptic enhancement may reflect properties characteristic of immature synaptic connections, and bursting activity in developing spinal neurons may promote functional maturation of the neuromuscular synapse. PMID- 10481008 TI - Recruitment of the auditory cortex in congenitally deaf cats by long-term cochlear electrostimulation. AB - In congenitally deaf cats, the central auditory system is deprived of acoustic input because of degeneration of the organ of Corti before the onset of hearing. Primary auditory afferents survive and can be stimulated electrically. By means of an intracochlear implant and an accompanying sound processor, congenitally deaf kittens were exposed to sounds and conditioned to respond to tones. After months of exposure to meaningful stimuli, the cortical activity in chronically implanted cats produced field potentials of higher amplitudes, expanded in area, developed long latency responses indicative of intracortical information processing, and showed more synaptic efficacy than in naive, unstimulated deaf cats. The activity established by auditory experience resembles activity in hearing animals. PMID- 10481009 TI - A chemical inhibitor of p53 that protects mice from the side effects of cancer therapy. AB - Chemotherapy and radiation therapy for cancer often have severe side effects that limit their efficacy. Because these effects are in part determined by p53 mediated apoptosis, temporary suppression of p53 has been suggested as a therapeutic strategy to prevent damage of normal tissues during treatment of p53 deficient tumors. To test this possibility, a small molecule was isolated for its ability to reversibly block p53-dependent transcriptional activation and apoptosis. This compound, pifithrin-alpha, protected mice from the lethal genotoxic stress associated with anticancer treatment without promoting the formation of tumors. Thus, inhibitors of p53 may be useful drugs for reducing the side effects of cancer therapy and other types of stress associated with p53 induction. PMID- 10481010 TI - A role for the proteasome in the light response of the timeless clock protein. AB - The cyclic expression of the period (PER) and timeless (TIM) proteins is critical for the molecular circadian feedback loop in Drosophila. The entrainment by light of the circadian clock is mediated by a reduction in TIM levels. To elucidate the mechanism of this process, the sensitivity of TIM regulation by light was tested in an in vitro assay with inhibitors of candidate proteolytic pathways. The data suggested that TIM is degraded through a ubiquitin-proteasome mechanism. In addition, in cultures from third-instar larvae, TIM degradation was blocked specifically by inhibitors of proteasome activity. Degradation appeared to be preceded by tyrosine phosphorylation. Finally, TIM was ubiquitinated in response to light in cultured cells. PMID- 10481011 TI - Fungal endophyte symbiosis and plant diversity in successional fields AB - Increasing evidence suggests that microbial interactions are important determinants of plant biodiversity. The hypothesis that fungal endophyte symbiosis reduces diversity in successional fields was tested by manipulating infection of tall fescue, the most abundant perennial grass in the eastern United States. Over a 4-year period, species richness declined and tall fescue dominance increased in infected plots relative to uninfected plots without differences in total productivity. A host-specific endophyte, with negligible biomass, altered plant community structure in this long-term field experiment and may be reducing plant diversity throughout its expanding range. PMID- 10481012 TI - Clonal interference and the evolution of RNA viruses. AB - In asexual populations, beneficial mutations that occur in different lineages compete with one another. This phenomenon, known as clonal interference, ensures that those beneficial mutations that do achieve fixation are of large effect. Clonal interference also increases the time between fixations, thereby slowing the adaptation of asexual populations. The effects of clonal interference were measured in the asexual RNA virus vesicular stomatitis virus; rates and average effects of beneficial mutations were quantified. PMID- 10481013 TI - High frequency of cryptic deleterious mutations in Caenorhabditis elegans. AB - Deleterious mutations with very small phenotypic effects could be important for several evolutionary phenomena, but the extent of their contribution has been unknown. Fitness effects of induced mutations in lines of Caenorhabditis elegans were measured using a system for which the number of deleterious point mutations in the DNA can be estimated. In fitness assays, only about 4 percent of the deleterious mutations fixed in each line were detectable. The remaining 96 percent, though cryptic, are significant for mutation load and, potentially, for the evolution of sex. PMID- 10481015 TI - The Drosophila poly(A)-binding protein II is ubiquitous throughout Drosophila development and has the same function in mRNA polyadenylation as its bovine homolog in vitro. AB - The poly(A)-binding protein II (PABP2) is one of the polyadenylation factors required for proper 3'-end formation of mammalian mRNAs. We have cloned Pabp2, the gene encoding the Drosophila homolog of mammalian PABP2, by using a molecular screen to identify new Drosophila proteins with RNP-type RNA-binding domains. Sequence comparison of PABP2 from Drosophila and mammals indicates that the most conserved domains are the RNA-binding domain and a coiled-coil like domain which could be involved in protein-protein interactions. Pabp2 produces four mRNAs which result from utilization of alternative poly(A) sites and encode the same protein. Using an antibody raised against Drosophila PABP2, we show that the protein accumulates in nuclei of all transcriptionally active cells throughout Drosophila development. This is consistent with a general role of PABP2 in mRNA polyadenylation. Analysis of Drosophila PABP2 function in a reconstituted mammalian polyadenylation system shows that the protein has the same functions as its bovine homolog in vitro : it stimulates poly(A) polymerase and is able to control poly(A) tail length. PMID- 10481014 TI - A piston model for transmembrane signaling of the aspartate receptor. AB - To characterize the mechanism by which receptors propagate conformational changes across membranes, nitroxide spin labels were attached at strategic positions in the bacterial aspartate receptor. By collecting the electron paramagnetic resonance spectra of these labeled receptors in the presence and absence of the ligand aspartate, ligand binding was shown to generate an approximately 1 angstrom intrasubunit piston-type movement of one transmembrane helix downward relative to the other transmembrane helix. The receptor-associated phosphorylation cascade proteins CheA and CheW did not alter the ligand-induced movement. Because the piston movement is very small, the ability of receptors to produce large outcomes in response to stimuli is caused by the ability of the receptor-coupled enzymes to detect small changes in the conformation of the receptor. PMID- 10481016 TI - Identification and characterization of nuclear matrix-attachment regions in the human serpin gene cluster at 14q32.1. AB - Matrix-attachment regions (MARs) are DNA elements that are defined by their abilities to bind to isolated nuclear matrices in vitro. The DNA sequences of different matrix-binding elements vary widely. The locations of some MARs at the ends of chromatin loops suggest that they may represent boundaries of individual chromatin domains. As such, MARs may play important roles in regulating transcription and chromatin structure. As a first step towards assessing the roles of MARs in these processes, we assayed DNA sequences from the human serine protease inhibitor (serpin) gene cluster at 14q32.1 for matrix-binding activity in vitro. This approximately 150 kb region contains the cell-specific genes encoding alpha1-anti-trypsin (alpha1AT) and corticosteroid-binding globulin (CBG), as well as an antitrypsin-related sequence termed ATR. A DNase I hypersensitive site (DHS) map of the locus has recently been described. We report here that the alpha1AT-ATR-CBG region contains five distinct MARs. There is a strong matrix-binding element approximately 16 kb upstream of alpha1AT; three MARs are between ATR and CBG and one MAR is within the CBG gene itself. These MARs were matrix-associated in all cell types examined. DNA sequencing indicated that the serpin MARs contained predominantly repetitive DNA, although the types of DNA repeats differed among the MARs. PMID- 10481017 TI - Plasmid DNA size does not affect the physicochemical properties of lipoplexes but modulates gene transfer efficiency. AB - Clinical applications of gene therapy mainly depend on the development of efficient gene transfer vectors. Large DNA molecules can only be transfected into cells by using synthetic vectors such as cationic lipids and polymers. The present investigation was therefore designed to explore the physicochemical properties of cationic lipid-DNA particles, with plasmids ranging from 900 to 52 500 bp. The colloidal stability of the lipoplexes formed by complexing lipopolyamine micelles with plasmid DNA of various lengths, depending on the charge ratio, resulted in the formation of three domains, respectively corresponding to negatively, neutrally and positively charged lipoplexes. Lipoplex morphology and structure were determined by the physicochemical characteristics of the DNA and of the cationic lipid. Thus, the lamellar spacing of the structure was determined by the cationic lipid and its spherical morphology by the DNA. The main result of this study was that the morphological and structural features of the lipopolyamine-DNA complexes did not depend on plasmid DNA length. On the other hand, their gene transfer capacity was affected by the size of plasmid DNA molecules which were sandwiched between the lipid bilayers. The most effective lipopolyamine-DNA complexes for gene transfer were those containing the shortest plasmid DNA. PMID- 10481018 TI - A neutralizing antibody against human DNA polymerase epsilon inhibits cellular but not SV40 DNA replication. AB - The contribution of human DNA polymerase epsilon to nuclear DNA replication was studied. Antibody K18 that specifically inhibits DNA polymerase activity of human DNA polymerase epsilon in vitro significantly inhibits DNA synthesis both when microinjected into nuclei of exponentially growing human fibroblasts and in isolated HeLa cell nuclei. The capability of this neutralizing antibody to inhibit DNA synthesis in cells is comparable to that of monoclonal antibody SJK 132-20 against DNA polymerase alpha. Contrary to the antibody against DNA polymerase alpha, antibody K18 against DNA polymerase epsilon did not inhibit SV40 DNA replication in vitro. These results indicate that DNA polymerase epsilon plays a role in replicative DNA synthesis in proliferating human cells like DNA polymerase alpha, and that this role for DNA polymerase epsilon cannot be modeled by SV40 DNA replication. PMID- 10481019 TI - The reactivity of the 2-deoxyribonolactone lesion in single-stranded DNA and its implication in reaction mechanisms of DNA damage and repair. AB - The formal C1'-oxidation product, 2-deoxyribonolactone, is formed as a result of DNA damage induced via a variety of agents, including gamma-radiolysis and the enediyne antitumor antibiotics. This alkaline labile lesion may also be an intermediate during DNA damage induced by copper-phenanthroline. Oligo nucleotides containing this lesion at a defined site were formed via aerobic photolysis of oligonucleotides containing a photolabile ketone, and were characterized by gel electrophoresis and electrospray mass spectrometry (ESI-MS). Treatment of oligo-nucleotides containing the lesion with secondary amines produces strand breaks consisting of 3'-phosphate termini, and products which migrate more slowly in polyacrylamide gels. MALDI-TOF mass spectrometry analysis indicates that the slower moving products are formal adducts of the beta elimination product resulting from 2-deoxyribonolactone and one molecule of amine. The addition of beta-mercapto-ethanol to the reaction mixture produces thiol adducts as well. The stability of these adducts suggests that they cannot be the labile species characterized by gel electrophoresis in copper phenanthroline-mediated strand scission. The characterization of these adducts by mass spectrometry also provides, by analogy, affirmation of proposals regarding the reactivity of nucleophiles with the beta-elimination product of abasic sites. Finally, the effects of this lesion and the various adducts on DNA repair enzymes are unknown, but their facile generation from oligonucleotides containing a photolabile ketone suggests that such issues could be addressed. PMID- 10481020 TI - Cloning and biochemical characterization of Bacillus subtilis YxiN, a DEAD protein specifically activated by 23S rRNA: delineation of a novel sub-family of bacterial DEAD proteins. AB - DEAD, DEAH and DExH proteins are involved in almost every facet of RNA biochemistry. Members of these protein families exhibit an RNA-dependent ATPase activity and some possess an ATP-dependent RNA helicase activity. Although genetic studies have identified specific functions for certain DEx(D)/(H)proteins from which an RNA substrate can be reasonably inferred, only DbpA from Escherichia coli has been shown to exhibit significant RNA specificity in vitro. Here we describe the characterization of YxiN from Bacillus subtilis, the second DEx(D)/(H)protein to show significant RNA specificity as an isolated, homogenous protein. The ATPase activity of YxiN, like that of DbpA, is stimulated by a 154 nt fragment of 23S rRNA. YxiN has a 2 nM apparent binding constant for this fragment, yet its ATPase activity shows 1800-fold RNA specificity. Along with the conserved motifs shared among all DEAD proteins, YxiN and DbpA have a conserved C terminal extension. This extension is highly conserved in several additional DEAD proteins. We propose that the C-terminus identifies a protein sub-family whose members bind 23S rRNA and that proteins of this family are likely to function in rRNA maturation/ribosome biogenesis or an unappreciated aspect of translation. PMID- 10481021 TI - Genome-wide expression profiling in Escherichia coli K-12. AB - We have established high resolution methods for global monitoring of gene expression in Escherichia coli. Hybridization of radiolabeled cDNA to spot blots on nylon membranes was compared to hybridization of fluorescently-labeled cDNA to glass microarrays for efficiency and reproducibility. A complete set of PCR primers was created for all 4290 annotated open reading frames (ORFs) from the complete genome sequence of E.coli K-12 (MG1655). Glass- and nylon-based arrays of PCR products were prepared and used to assess global changes in gene expression. Full-length coding sequences for array printing were generated by two step PCR amplification. In this study we measured changes in RNA levels after exposure to heat shock and following treatment with isopropyl-beta-D thiogalactopyranoside (IPTG). Both radioactive and fluorescence-based methods showed comparable results. Treatment with IPTG resulted in high level induction of the lacZYA and melAB operons. Following heat shock treatment 119 genes were shown to have significantly altered expression levels, including 35 previously uncharacterized ORFs and most genes of the heat shock stimulon. Analysis of spot intensities from hybridization to replicate arrays identified sets of genes with signals consistently above background suggesting that at least 25% of genes were expressed at detectable levels during growth in rich media. PMID- 10481022 TI - NMR studies on truncated sequences of human telomeric DNA: observation of a novel A-tetrad. AB - The structure of the telomeric DNA has been a subject of extensive investigation in recent years due to the realization that it has important functional roles to play in vivo and the observations that truncated telomeric sequences exhibit a great variety of 3D structures in aqueous solutions. In this context, we describe here NMR structural studies on two truncated human telomeric DNA sequences, d AG(3)T and d-TAG(3)T in solutions containing K(+)ions. The G(3)stretches in both the oligonucleotides were seen to form parallel-stranded quadruplexes. However, the AG(3)segment as a whole, had different structural characteristics. The structure of d-AG(3)T revealed the formation of a novel A-tetrad, which was not seen in d-TAG(3)T. The A's in the tetrad had syn glycosidic conformation as opposed to the anti conformation of the G's in the G-tetrads. The A-tetrad stacked well over the adjacent G-tetrad and the twist angle at this step was smaller in d-AG(3)T than in d-TAG(3)T. These observations are expected to be significant from the point of view of structural diversity and recognition in telomeres. PMID- 10481023 TI - The stability of DNA triplexes inside cells as studied by iodine-125 radioprinting. AB - We studied the stability of a DNA triplex resulting from the binding of a 38 nt long purine motif triplex-forming oligonucleotide (TFO) to a covalently closed plasmid containing a target sequence from the human HPRT gene. Our in vitro experiments showed that the triplex formed at plasmid and TFO concentrations as low as 10(-9)M. Once formed, the triplex was remarkably stable and could withstand 10 min incubation at 65 degrees C. We next delivered these TFO-plasmid complexes into cultured human cells. To monitor the TFO-plasmid complexes inside cells we applied a new technique that we call 'radioprinting'. Because the TFO was(125)I labeled, we could quantitatively monitor the triplexes by measuring(125)I-induced DNA strand breaks in the target plasmid sequence. We found that the triplexes remain stable inside the cells for at least 48 h. Based on these findings we propose using TFO for indirect labeling of intact plasmid DNA. As a demonstration, we show that the intracellular distribution of a fluorescein-labeled TFO was different when it was liposome-delivered into cultured human cells alone or in a complex with the plasmid. In the latter case, the fluorescence was detected in nearly all the cells while detection of the plasmid by use of a marker gene (beta-galactosidase) revealed expression of the gene in only half of the cells. PMID- 10481024 TI - Fold-back structures at the distal end influence DNA slippage at the proximal end during mononucleotide repeat expansions. AB - Polymerase slippage during DNA synthesis by the Klenow fragment of DNA polymerase across A, C, G and T repeats (30 bases) has been studied. Within minutes, duplexes that contain only repeats (30 bp) expand dramatically to several hundred base pairs long. Rate comparisons in a repeat duplex when one strand was expanded as against that when both strands were expanded suggest a model of migrating hairpin loops which in the latter case coalesce into a duplex. Moreover, slippage (at the proximal or 3'-end) is subject to positive and negative effects from the 5'-end (distal) of the same strand. Growing T and G strands generate T.A:T and G G:C motif fold-back structures at the distal end that hamper slippage at the proximal end. On the other hand, growing tails at the distal end upon annealing with excess complementary template accentuates proximal slippage several-fold. PMID- 10481025 TI - In vitro transposition of Tn552: a tool for DNA sequencing and mutagenesis. AB - We have explored the potential of the Tn 552 in vitro transposition reaction as a genetic tool. The reaction is simple (requiring a single protein component), robust and efficient, readily producing insertions into several percent of target DNA. Most importantly, Tn 552 insertions in vitro appear to be essentially random. Extensive analyses indicate that the transposon exhibits no significant regional or sequence specificity for target DNA and leaves no discernible 'cold' spots devoid of insertions. The utility of the in vitro reaction for DNA sequencing was demonstrated with a cosmid containing the Mycobacterium smegmatis recBCD gene cluster. The nucleotide sequence of the entire operon was determined using 71 independent Tn 552 insertions, which generated over 13.5 kb of unique sequence and simultaneously provided a comprehensive collection of insertion mutants. The relatively short ends of Tn 552 make construction of novel transposons a simple process and we describe several useful derivatives. The data presented suggest that Tn 552 transposition is a valuable addition to the arsenal of tools available for molecular biology and genomics. PMID- 10481026 TI - Comparative analysis of splicing of the complete set of chloroplast group II introns in three higher plant mutants. AB - The barley mutant albostrians and the maize mutants crs1 and crs2 are defective in the splicing of various plastid group II introns. By analysing tRNA precursors and several mRNAs not previously examined, the investigation of in vivo splicing defects in these mutants has been completed. The albostrians mutation causes the loss of plastid ribosomes resulting secondarily in a disruption of splicing of all subgroup IIA introns in the chloroplast. Thus MatK, the only putative chloroplast intron-specific maturase of higher plants, might have evolved to function in splicing of multiple introns. We show that in the case of tRNA Ala(UGC)the first step of splicing is affected, as suggested by the absence of lariat molecules. Thus the plastid-encoded splicing factor lacking in albostrians must participate in the formation of the catalytically active structure. In contrast, a mutation in the nuclear gene crs1 prevents splicing of only one intron but causes specific additional effects as precursor transcripts for tRNA Ile(GAU), tRNA-Ala(UGC), tRNA-Lys(UUU)and tRNA-Val(UAC), but not tRNA-Gly(UCC), have significantly enhanced steady-state levels in this mutant. Our data provide evidence for a variety of splicing factors and pathways in the chloroplast, some encoded by nuclear and some by chloroplast genes, and possibly for a dual function of some of these factors. PMID- 10481027 TI - DNA bending by photolyase in specific and non-specific complexes studied by atomic force microscopy. AB - Specific and non-specific complexes of DNA and photolyase are visualised by atomic force microscopy. As a substrate for photolyase a 1150 bp DNA restriction fragment was UV-irradiated to produce damaged sites at random positions. Comparison with a 735 bp undamaged DNA fragment made it possible to separate populations of specific and non-specific photolyase complexes on the 1150 bp fragment, relieving the need for highly defined substrates. Thus it was possible to compare DNA bending for specific and non-specific interactions. Non-specific complexes show no significant bending but increased rigidity compared to naked DNA, whereas specific complexes show DNA bending of on average 36 degrees and higher flexibility. A model obtained by docking shows that photolyase can accommodate a 36 degrees bent DNA in the vicinity of the active site. PMID- 10481028 TI - Presetting of chromatin structure and transcription factor binding poise the human GADD45 gene for rapid transcriptional up-regulation. AB - GADD45 has been suggested to coordinate cell cycle regulation with the repair of DNA damage following ionizing radiation (IR). Although the GADD45 gene is transcriptionally up-regulated in response to IR, alterations in in vivo transcription factor (TF) binding or chromatin structure associated with up regulation have not been defined. To understand how chromatin structure might influence TF binding and GADD45 up-regulation, key regulatory regions of the gene were identified by in vivo DNase I hypersensitivity (HS) analysis. Chromatin structure and in vivo TF binding in these regions were subsequently monitored in both non-irradiated and irradiated human ML-1 cells. In non-irradiated cells expressing basal levels of GADD45, the gene exhibited a highly organized chromatin structure with distinctly positioned nucleosomes. Also identified in non-irradiated cells were DNA-protein interactions at octamer binding motifs and a CCAAT box in the promoter and at consensus binding sites for AP-1 and p53 within intron 3. Upon irradiation and a subsequent 15-fold increase in GADD45 mRNA levels, neither the chromatin structure nor the pattern of TF binding in key regulatory regions was altered. These results suggest that the GADD45 gene is poised for up-regulation and can be rapidly induced independent of gross changes in chromatin structure or TF binding. PMID- 10481029 TI - In vivo oligo(A) insertions in phage MS2: role of Escherichia coli poly(A) polymerase. AB - Previously we introduced an RNase III site into the genome of RNA phage MS2 by extending a hairpin with a perfect 18 bp long stem. One way in which the phage escaped from being killed by RNase III cleavage was to incorporate uncoded A residues on either side of the stem. This oligo(A) stretch interrupts the perfect stem that forms the RNase III site and thus confers resistance. In this paper we have analyzed the origin of these uncoded adenosines. The data strongly suggest that they are added by the host enzyme poly(A) polymerase. Apparently the 3'-OH created by RNase III cleavage becomes a substrate for poly(A) polymerase. Subsequently, MS2 replicase makes one contiguous copy from the two parts of the genome RNA. The evolutionary conversion from RNase III sensitivity to resistance provides a large spectrum of solutions that could be an important tool to understand what essentially constitutes an RNase III site in vivo. PMID- 10481031 TI - Heuristic approach to deriving models for gene finding. AB - Computer methods of accurate gene finding in DNA sequences require models of protein coding and non-coding regions derived either from experimentally validated training sets or from large amounts of anonymous DNA sequence. Here we propose a new, heuristic method producing fairly accurate inhomogeneous Markov models of protein coding regions. The new method needs such a small amount of DNA sequence data that the model can be built 'on the fly' by a web server for any DNA sequence >400 nt. Tests on 10 complete bacterial genomes performed with the GeneMark.hmm program demonstrated the ability of the new models to detect 93.1% of annotated genes on average, while models built by traditional training predict an average of 93.9% of genes. Models built by the heuristic approach could be used to find genes in small fragments of anonymous prokaryotic genomes and in genomes of organelles, viruses, phages and plasmids, as well as in highly inhomogeneous genomes where adjustment of models to local DNA composition is needed. The heuristic method also gives an insight into the mechanism of codon usage pattern evolution. PMID- 10481030 TI - Comparison of five methods for finding conserved sequences in multiple alignments of gene regulatory regions. AB - Conserved segments in DNA or protein sequences are strong candidates for functional elements and thus appropriate methods for computing them need to be developed and compared. We describe five methods and computer programs for finding highly conserved blocks within previously computed multiple alignments, primarily for DNA sequences. Two of the methods are already in common use; these are based on good column agreement and high information content. Three additional methods find blocks with minimal evolutionary change, blocks that differ in at most k positions per row from a known center sequence and blocks that differ in at most k positions per row from a center sequence that is unknown a priori. The center sequence in the latter two methods is a way to model potential binding sites for known or unknown proteins in DNA sequences. The efficacy of each method was evaluated by analysis of three extensively analyzed regulatory regions in mammalian beta-globin gene clusters and the control region of bacterial arabinose operons. Although all five methods have quite different theoretical underpinnings, they produce rather similar results on these data sets when their parameters are adjusted to best approximate the experimental data. The optimal parameters for the method based on information content varied little for different regulatory regions of the beta-globin gene cluster and hence may be extrapolated to many other regulatory regions. The programs based on maximum allowed mismatches per row have simple parameters whose values can be chosen a priori and thus they may be more useful than the other methods when calibration against known functional sites is not available. PMID- 10481032 TI - The inefficient replication origin from yeast ribosomal DNA is naturally impaired in the ARS consensus sequence and in DNA unwinding. AB - Ribosomal DNA (rDNA) replication origins of Saccharomyces cerevisiae are known to function inefficiently, both in the context of the tandem rDNA repeats in the chromosome and as single copy autonomously replicating sequences (ARSs) in plasmids. Here we examined components of the rDNA ARS that might contribute to inefficient extrachromosomal replication. Like the efficient H4 ARS, the rDNA ARS requires a match to the 11 bp ARS consensus sequence (ACS) and a broad non conserved region that may contain multiple elements, including a DNA unwinding element (DUE). Using a single-strand-specific nuclease hypersensitivity assay and by determining the superhelical density required for stable DNA unwinding, we found that the DNA of the rDNA ARS is not as easily unwound as the H4 ARS. Unwinding of the rDNA ARS required additional energy, similar to the unwinding of mutations in the H4 ARS that stabilize the double helix in the DUE region and impair replication. In vivo extrachromosomal replication of the rDNA ARS was cold sensitive, like H4 ARS mutants that require additional energy to unwind the DUE region but unlike the easily unwound, wild-type H4 ARS. Impairment of replication function at reduced temperature suggests that the elevated energy requirement for DNA unwinding inherent in the wild-type rDNA ARS contributes to inefficient replication function. We also examined the essential ACS match in the rDNA ARS, which is known to be imperfect at one position. A point mutation in the essential ACS that corrects the imperfect match increased the efficiency of extrachromosomal replication. Our results reveal that the essential ACS element and DNA unwinding in the rDNA ARS are naturally impaired, suggesting that inefficient function of the rDNA replication origin has a biological purpose. PMID- 10481033 TI - The non-enzymatic hydrolysis of oligoribonucleotides VI. The role of biogenic polyamines. AB - Single-stranded oligoribonucleotides containing UA and CA phosphodiester bonds can be hydrolyzed specifically under non-enzymatic conditions in the presence of spermidine, a biogenic amine found in a wide variety of organisms. In the present study, the rate of oligonucleotide and tRNA(i)(Met)hydrolysis was measured in the presence of spermidine and other biogenic amines. It was found that spermine [H(3)N(+)(CH(2))(3)(+)NH(2)(CH(2))(4)(+)NH(2)(CH(2))(3)(+)NH(3)] and putrescine [H(3)N(+)(CH(2))(4)(+)NH(3)] can replace spermidine [H(3)N(+) (CH(2))(4)(+)NH(2)(CH(2))(3)(+)NH(3)] to induce the hydrolysis. For all three polyamines, a bell-shaped cleavage rate versus concentration relationship was observed. The maximum rate of hydrolysis was achieved at 0.1, 1.0 and 10 mM spermine, spermidine and putrescine, respectively. Moreover, we found that the hydrolysis requires at least two linked amino groups since two aminoalcohols, 2 aminoethanol and 3-aminopropanol, were not able to induce the cleavage of the phospho-diester bond. The optimal cleavage rate of the oligo-ribonucleotides was observed when amino groups were separated by tri- or tetramethylene linkers. The methylation of the amino groups reduced the ability of diamines to induce oligoribonucleotide hydrolysis. Non-enzymatic cleavage of tRNA(i)(Met)from Lupinus luteus and tRNA(i)(Met)from Escherichia coli demonstrate that both RNAs hydrolyze as expected from principles derived from oligoribonucleotide models. PMID- 10481034 TI - Structural characterisation of a uracil containing hairpin DNA by NMR and molecular dynamics. AB - Three-dimensional (3D) structure of a hairpin DNA d-CTAGAGGATCCTTTUGGATCCT (22mer; abbreviated as U4-hairpin), which has a uracil nucleotide unit at the fourth position from the 5' end of the tetra-loop has been solved by NMR spectroscopy. The(1)H resonances of this hairpin have been assigned almost completely. NMR restrained molecular dynamics and energy minimisation procedures have been used to describe the 3D structure of the U4 hairpin. This study establishes that the stem of the hairpin adopts a right handed B-DNA conformation while the T(12)and U(15)nucleotide stack upon 3' and 5' ends of the stem, respectively. Further, T(14)stacks upon both T(12)and U(15)while T(13)partially stacks upon T(14). Very weak stacking interaction is observed between T(13)and T(12). All the individual nucleotide bases adopt ' anti ' conformation with respect to their sugar moiety. The turning phosphate in the loop is located between T(13)and T(14). The stereochemistry of U(15)mimics the situation where uracil would stack in a B-DNA conformation. This could be the reason as to why the U4-hairpin is found to be the best substrate for its interaction with uracil DNA glycosylase (UDG) compared to the other substrates in which the uracil is at the first, second and third positions of the tetra-loop from its 5' end, as reported previously. PMID- 10481035 TI - Accurate selection of a 5' splice site requires sequences within fibronectin alternative exon B. AB - Inclusion of fibronectin alternative exon B in mRNA is developmentally regulated. Here we demonstrate that exon B contains two unique purine-rich sequence tracts, PRE1 and PRE2, that are important for proper 5' splice site selection both in vivo and in vitro. Targeted mutations of both PREs decreased the inclusion of exon B in the mRNA by 50% in vivo. Deletion or mutation of the PREs reduced removal of the downstream intron, but not the upstream intron, and induced the activation of cryptic 5' splice sites in vitro. PRE-mediated 5' splice selection activity appears sensitive to position and sequence context. A well characterized exon sequence enhancer that normally acts on the upstream 3' splice site can partially rescue proper exon B 5' splice site selection. In addition, we found that PRE 5' splice selection activity was preserved when exon B was inserted into a heterologous pre-mRNA substrate. Possible roles of these unique activities in modulating exon B splicing are considered. PMID- 10481036 TI - Differential subtraction chain, a method for identifying differences in genomic DNA and mRNA. AB - Identifying the genetic differences between two organisms or cell types has been a major goal in modern biomedical research. Recently, we developed a novel methodology that can rapidly identify the differences between two populations of DNA. This method, termed 'differential subtraction chain' (DSC), is based on a novel 'negative amplification' strategy that converts (amplifiable) tester sequences to counterpart (unamplifiable) driver sequences. The result is a double exponential elimination of amplifiable sequences in the testers, while preserving the sequences in the testers that have no counterpart in the drivers. We applied this methodology to the genome of a glioblastoma cell line. A homozygous deletion was rapidly identified. We extended this technique to identifying the unique sequences in mRNA. Two CDC25 transgene fragments were quickly identified in a cdc25B transgenic mouse. We also applied this methodology to systems with profound differences in mRNA expression. In a 'prostate epithelia subtracting blood cells' DSC reaction, a sample of unique gene fragments which are absent in the prostate but present in the blood were identified. Lastly, we detected rare (1 virus/100 cells) Herpes simplex virus type 2 (HSV-2) sequences in a tissue culture, indicating good sensitivity of this methodology. Overall, DSC represents a fast, efficient and sensitive method for identifying differences in genomic DNA and mRNA and can be easily applied in a variety of biological systems. PMID- 10481037 TI - Signal amplification system for DNA hybridization assays based on in vitro expression of a DNA label encoding apoaequorin. AB - The development of hybridization assays based on an apoaequorin-encoding DNA label is reported. The constructed label contains the T7 RNA polymerase promoter, the apoaequorin coding sequence and a downstream (dA/dT)(30). In the captured target configuration, biotinylated target DNA (233 bp) was captured on streptavidin-coated microtiter wells and hybridized to a poly(dT)-tailed detection probe. In the sandwich-type assay, the target DNA was hybridized simultaneously with an immobilized capture probe (through biotin/streptavidin) and a poly(dT)-tailed detection probe. In both configurations, the hybrids were reacted with poly(dA)-tailed apoaequorin DNA. The DNA label was subjected to in vitro transcription/translation to produce apoaequorin, which was converted to active aequorin in the reaction mixture. Generated aequorin was determined by its characteristic Ca(2+)-triggered bioluminescence. Each DNA label was estimated to produce 156 aequorin molecules. As low as 0.25 and 0.5 amol of target DNA were detected with the sandwich-type and captured target hybridization assays, respectively, with a linear range spanning four orders of magnitude. In comparison, captured target hybridization assays using photoprotein aequorin or firefly luciferase-encoding DNA labels were able to detect 25 and 20.5 amol of target DNA, respectively. The dramatic improvement in sensitivity observed with the proposed systems is attributed to amplification introduced by in vitro expression of apoaequorin DNA into multiple active aequorin molecules. PMID- 10481038 TI - Enzyme-free cloning: a rapid method to clone PCR products independent of vector restriction enzyme sites. AB - We describe a simple method for the cloning of PCR products without the need for post-amplification enzymatic treatment. Tailed PCR primer sets are used to create complementary staggered overhangs on both insert and vector by a post-PCR denaturation-hybridisation reaction. The single-stranded overhangs are designed to allow directional cloning in a ligase-free manner. This 'enzyme-free cloning' procedure is highly efficient, and is not constrained by the need for the presence of suitable restriction enzyme sites within the plasmid vector. The avoidance of post-amplification enzymatic procedures makes the technique rapid and reliable, avoiding the need for multiple sub-cloning steps. PMID- 10481039 TI - Gene targeting restricted to mouse striated muscle lineage. AB - Spatially and temporally regulated somatic mutations can be achieved by using the Cre/LoxP recombination system of bacteriophage P1. In order to develop gene knockouts restricted to striated muscle, we generated a transgenic mouse line expressing Cre recombinase under the control of the human alpha-skeletal actin promoter. Specific excision of a loxP-flanked gene was demonstrated in striated muscle, heart and skeletal muscle, in a pattern very similar to the expression of the endogenous alpha-skeletal actin gene. Therefore, the reported transgenic line can be used to target inactivation or activation of a given gene to the skeletal muscle lineage. PMID- 10481040 TI - Direct sequencing of RAPD fragments using 3'-extended oligonucleotide primers and dye terminator cycle-sequencing. AB - Random amplified polymorphic DNA (RAPD) markers are used widely to develop high resolution genetic maps and for genome fingerprinting. Typically, single oligomers of approximately 10 nucleotides are used to PCR amplify characteristic RAPD marker fragments. We describe an efficient method for the direct end sequencing of gel-purified RAPD fragments using one primer from a set of four 3' terminal extended (A, T, C or G) oligonucleotides, identical to the RAPD primer but for the single nucleotide extension. Strand-specific DNA sequence could be independently read from each of the RAPD fragments without recourse to strand separation or fragment cloning. Informative RAPD fragments could be readily converted into mapped STS or SCAR loci using this technology. The 3'-extended primers may also be used to amplify independent genomic RAPD markers. PMID- 10481041 TI - Similarities and dissimilarities in the structure-function relation between the cytochrome c oxidase from bovine heart and from Paracoccus denitrificans as revealed by FT-IR difference spectroscopy. AB - The redox dependent changes in the cytochrome c oxidase from bovine heart were studied with a combined electrochemical and FT-IR spectroscopic approach. A direct comparison to the electrochemically induced FT-IR difference spectra of the cytochrome c oxidase from Paracoccus denitrificans reveals differences in the structure and intensity of vibrational modes. These differences are partially attributed to interactions of subunits influencing the heme and protein modes. In the spectral regions characteristic for v(C=O) and v(COO-)s/as modes of protonated and deprotonated Asp and Glu residues, additional signals at 1736, 1602 and 1588 cm-1 are observed. On this basis, the possible involvement of Asp 51, a residue specifically conserved in mammalian oxidase and previously proposed to show redox depended conformational changes in the respective X-ray structures, is critically discussed. PMID- 10481042 TI - Light-induced degradation of cytochrome b559 during photoinhibition of the photosystem II reaction center. AB - The behaviour of cytochrome (cyt) b559 during acceptor- and donor-side photoinhibition has been investigated in oxygen-evolving and non-evolving photosystem II (PSII) membranes. Strong illumination at 20 degrees C under aerobiosis induced a strong decrease in the absorbance of the cyt b559 alpha-band in the two preparations. This absorbance decline was observed only in non-oxygen evolving PSII samples when illumination was performed under aerobiosis but at 4 degrees C, or under anaerobiosis at 20 degrees C. These results suggest that acceptor-side photoinhibition induces the degradation of cyt b559 by a mechanism related to an enzymatic reaction mediated by singlet oxygen. Donor-side photoinhibition may induce, however, a non-enzymatic photocleavage of the protein. PMID- 10481043 TI - Plausible phosphoenolpyruvate binding site revealed by 2.6 A structure of Mn2+ bound phosphoenolpyruvate carboxylase from Escherichia coli. AB - We have determined the crystal structure of Mn2+-bound Escherichia coli phosphoenolpyruvate carboxylase (PEPC) using X-ray diffraction at 2.6 A resolution, and specified the location of enzyme-bound Mn2+, which is essential for catalytic activity. The electron density map reveals that Mn2+ is bound to the side chain oxygens of Glu-506 and Asp-543, and located at the top of the alpha/beta barrel in PEPC. The coordination sphere of Mn2+ observed in E. coli PEPC is similar to that of Mn2+ found in the pyruvate kinase structure. The model study of Mn2+-bound PEPC complexed with phosphoenolpyruvate (PEP) reveals that the side chains of Arg-396, Arg-581 and Arg-713 could interact with PEP. PMID- 10481044 TI - Light-induced cytochrome P450-dependent enzyme in indole alkaloid biosynthesis: tabersonine 16-hydroxylase. AB - Vinblastine and vincristine are two medically important bisindole alkaloids from Catharanthus roseus (Madagascar periwinkle). Attempts at production in cell cultures failed because a part of the complex pathway was not active, i.e. from tabersonine to vindoline. It starts with tabersonine 16-hydroxylase (T16H), a cytochrome P450-dependent enzyme. We now show that T16H is induced in the suspension culture by light and we report the cloning of the cDNA. The enzyme was expressed in Escherichia coli as translational fusion with the P450 reductase from C. roseus, and the reaction product was identified by mass spectrometry. The protein (CYP71D12) shares 47-52% identity with other members of the CYP71D subfamily with unknown function. The induction by light was strongly enhanced by a nutritional downshift (transfer into 8% aqueous sucrose). We discuss the possibility that the entire pathway to bisindoles can be expressed in suspension cultures. PMID- 10481045 TI - Higher efficiency of the liver phosphorylative system in diabetic Goto-Kakizaki (GK) rats. AB - Liver mitochondrial bioenergetics of Goto-Kakizaki (GK) rats (a model of non insulin dependent diabetes mellitus) reveals a Delta Psi upon energization with succinate significantly increased relatively to control animals. The repolarization rate following ADP phosphorylation is also significantly increased in GK mitochondria in parallel with increased ATPase activity. The increase in the repolarization rate and ATPase activity is presumably related to an improved efficiency of F(0)F(1)-ATPase, either from a better phosphorylative energy coupling or as a consequence of an enlarged number of catalytic units. Titrations with oligomycin indicate that diabetic GK liver mitochondria require excess oligomycin pulses to completely abolish phosphorylation, relative to control mitochondria. Therefore, accepting that the number of operational ATP synthase units is inversely proportional to the amount of added oligomycin, it is concluded that liver mitochondria of diabetic GK rats are provided with extra catalytic units relative to control mitochondria of normal rats. Other tissues (kidney, brain and skeletal muscle) were evaluated for the same bioenergetic parameters, confirming that this feature is exclusive to liver from diabetic GK rats. PMID- 10481046 TI - Gangliosides GM1 and GD1b are not polarized in mature hippocampal neurons. AB - Analysis of the binding of cholera toxin to ganglioside GM1 in both living and fixed neurons, and comparison with the distribution of defined axonal and dendritic proteins, demonstrates that ganglioside GM1 is distributed in a non polarized manner over the axonal and dendritic plasma membranes of mature, cultured hippocampal neurons. Likewise, ganglioside GD1b is also distributed in a non-polarized manner. These results suggest that a recent report [Ledesma, M.D. et al. EMBO J. 18 (1999) 1761-1771] proposing that ganglioside GM1 is highly enriched on the axonal versus dendritic membrane of hippocampal neurons may need to be re-evaluated. PMID- 10481047 TI - Determination of carbohydrate-deficient transferrin separated by lectin affinity chromatography for detecting chronic alcohol abuse. AB - Carbohydrate-deficient transferrin (CDT) has been established as a valuable biological marker for detecting chronic alcohol abuse. To improve the diagnostic efficiency, we studied new CDT determination procedures involving the use of lectin affinity chromatography with Allomyrina dichotoma agglutinin (allo A) and Trichosanthes japonica agglutinin I (TJA-I) to isolate the CDT isoforms CDT-allo A and CDT-TJA, respectively. These procedures, based on detection of the CDT-allo A and CDT-TJA isoforms in sera, showed high sensitivity (100% and 98%, respectively) and high specificity (93% and 85%, respectively). These results demonstrate that the new procedures involving the use of lectin affinity chromatography are more useful for isolating markers in the CDT test than the conventional charge-based separation method. PMID- 10481048 TI - HtpG is essential for the thermal stress management in cyanobacteria. AB - The heat shock protein (Hsp) HtpG is a member of the Hsp90 protein family. We cloned a single-copy gene encoding a homologue of HtpG from the unicellular cyanobacterium Synechococcus sp. PCC 7942. Sequence alignment with HtpGs from other prokaryotes revealed unique features in the cyanobacterial HtpG primary sequence. A monocistronic mRNA of the htpG gene increased transiently in response to heat shock. In order to elucidate the role of HtpG in vivo, we inactivated the htpG gene by targeted mutagenesis. Although the mutation did not affect the photoautotrophic growth at 30 and 42 degrees C, the mutant cells were unable to grow at 45 degrees C. They lost both basal and acquired thermotolerances. These results indicate that HtpG plays an essential role for the thermal stress management in cyanobacteria, the first such an example for either a photosynthetic or a prokaryotic organism. PMID- 10481049 TI - Characterization of a plant mitochondrial active chromosome. AB - A method is presented for the partial purification of a plant mitochondrial active chromosome (MAC). This method is based on the presence of the mitochondrial chromosome in the insoluble mitochondrial fraction which allows for its rapid purification from the bulk of detergent-solubilized proteins by ultra centrifugation. The resuspended MAC carrying DNA and RNA-binding proteins retains DNA synthesis and transcription activities comparable to the ones found in isolated mitochondria. In comparison, tRNA-nucleotidyl terminal transferase taken as an example of RNA modifying activities remains in the soluble fraction. MAC purification is proposed as a rapid and efficient first step in the purification of DNA-binding proteins involved in DNA replication and transcription. PMID- 10481050 TI - Functional reconstitution of beta-glucan elicitor-binding activity upon incorporation into lipid vesicles. AB - In temperature-induced Triton X-114 phase separation experiments the beta-glucan elicitor-binding site from soybean (Glycine max L.) root membranes was identified as (a) hydrophobic membrane protein(s). The Zwittergent 3-12-solubilized beta glucan-binding proteins were incorporated into lipid vesicles by the detergent dilution procedure. Reconstituted binding proteins were functional in that binding of the hepta-beta-glucoside ligand was saturable, reversible and of high affinity (K(d)=6-7 nM). Competition studies using beta-glucans with different degrees of polymerization (DP 7-15; DP 15-25) showed effective displacement of the radioligand from the binding site whereas beta-glucan fragments with DP <7 were ineffective. The total amount of reconstituted binding activity was dependent on the acyl chain length of the phospholipids used for the reconstitution with a preference for decanoic (C10) and dodecanoic (C12) chains. Restored ligand binding was maximally 37% as compared to the former detergent solubilized binding activity. The presence of a lipid environment stabilized the purified beta-glucan-binding proteins. PMID- 10481051 TI - Glutathione peroxidase-like activity of caeruloplasmin as an important lung antioxidant. AB - The copper-containing plasma protein caeruloplasmin (Cp) has been shown to possess several oxidase activities, but with the exception of its ferrous ion oxidising (ferroxidase) activity which so far appear to be of minor biological relevance. Recently, Kim and colleagues (Kim et al. (1998) FEBS Lett. 431, pp. 473-475) observed that Cp can catalytically remove hydrogen peroxide in the presence of thiols. Here, we show that Cp can remove both hydrogen peroxide and lipid hydroperoxides at physiologically relevant concentrations of reduced glutathione known to be present in lung and lung lining fluid. The glutathione peroxidase-like activity of Cp together with its ferroxidase activity would completely remove the primary reactants required for both Fenton chemistry and lipid peroxidation. PMID- 10481053 TI - Characterization of testicular mouse glucosamine 6-phosphate deaminase (GNPDA). AB - Mammalian glucosamine 6-phosphate deaminase (GNPDA) was first detected in hamster spermatozoa. To further elucidate its role, we have cloned mouse GNPDA and produced a polyclonal rabbit anti-GNPDA antibody. This antibody recognized a 33 kDa protein in soluble extracts from mouse brain, liver, kidney, muscle, ovary, testis and sperm. Immunofluorescent analysis of the localization of GNPDA in male reproductive tissue revealed its presence in spermatids and in spermatozoa. In spermatids, GNPDA localized close to the developing acrosome vesicle and in spermatozoa close to the acrosomal region. Following the induction of the acrosome reaction, GNPDA fluorescence in spermatozoa was either reduced or GNPDA was absent. These data suggest that GNPDA might play a role in the acrosome reaction. PMID- 10481052 TI - Transient Ca2+-dependent activation of ERK1 and ERK2 in cytotoxic responses induced by maitotoxin in breast cancer cells. AB - Treatment of MCF-7 breast cancer cells with the marine toxin maitotoxin (MTX) induces cell death. The cytotoxic effects are clearly detectable within 2-4 h after cell treatment with 10(-10)-10(-9) M concentrations of MTX. The response was found to depend on extracellular Ca2+, inasmuch as cell death was prevented when culture dishes received MTX, following addition of EGTA. MTX caused transient phosphorylation of extracellular signal-regulated kinase isoforms 1 and 2 (ERK1 and ERK2) mitogen-activated protein kinase isoforms in MCF-7 cells, which was maximal 15 min after toxin addition to culture vessels. The effect was dependent on influx of extracellular Ca2+, as it was abolished by EGTA, and was induced by ionophores, such as A23187 and ionomycin. Our findings show that signaling pathways involving Ca2+ ions may cause activation of ERK1 and ERK2 in cell death responses. PMID- 10481054 TI - Thermodynamics of nucleotide binding to NBS-I of the Bacillus subtilis preprotein translocase subunit SecA. AB - SecA is the dissociatable nucleotide and preprotein binding subunit of the bacterial translocase. The thermodynamics of nucleotide binding to soluble SecA at nucleotide binding site I were determined by isothermal titration calorimetry. Binding of ADP and non-hydrolyzable ATPgammaS is enthalpy-driven (DeltaH(0) of 14.44 and -5.56 kcal/mol, respectively), but is accompanied by opposite entropic contributions (DeltaS(0) of -18.25 and 9.55 cal/mol/K, respectively). ADP binding results in a large change in the heat capacity of SecA (DeltaC(p)=-780 cal/mol/K). It is suggested that ADP binding promotes the interaction between the two thermodynamically discernible domains of SecA which is accompanied by a shielding of hydrophobic surface from solvent. PMID- 10481055 TI - Inhibition of infection of incoming HIV-1 virus by RNA-cleaving DNA enzyme. AB - Nine different DNA enzymes (DzV3-n, n=1-9) targeting the V3 loop region of HIV-1 HXB2 were synthesized. One of those, DzV3-9, efficiently cleaved the target in the conserved sequence in the RNA transcript in vitro. DzV3-9 was stable in the cells and inhibited replication of both NL432 and SF162 strains in U87 cells expressing CD4 and co-receptors. The inhibitory effect of DNAzyme on incoming HIV 1 was also demonstrated with pseudotype virions generated by NL432-based luciferase reporter genes. Thus, an efficient, stable DNAzyme against a functionally important region of HIV-1 was identified, and it may be useful for prevention of HIV-1 infection. PMID- 10481056 TI - Difference in induction of uncoupling protein genes in adipose tissues between young and old rats during cold exposure. AB - Induction of uncoupling protein (UCP) genes in adipose tissues from young and old rats exposed to cold was compared. UCP1 mRNA expression in brown adipose tissue (BAT) was enhanced in both young and old rats after cold exposure, but the expression was downregulated at 72 h after the exposure only in the young rats. The UCP2 gene was induced notably in BAT of young rats instead of the downregulation of the UCP1 gene, whereas the induction in old rats was almost blunted. The pattern of UCP3 expression was similar to that of UCP1 expression in each group. The effect of cold exposure on the expression of UCP2 genes was also observed in white adipose tissue from the young rats. These results indicate a change in induction of UCP genes in adipose tissues with aging. PMID- 10481058 TI - Proteolytic cleavage of beta-catenin by caspases: an in vitro analysis. AB - Cleavage of structural proteins by caspases has been associated with the severe morphological changes occurring during the apoptotic process. One of the proteins regulating the connection of the actin filament with cadherins in a cell-cell adhesion complex is beta-catenin. During apoptosis, both an N-terminal and a small C-terminal part are removed from beta-catenin. Removal of the N-terminal part may result in a disconnection of the actin filament from a cadherin cell cell adhesion complex. We demonstrate that caspase-8, -3 and -6 directly proteolyse beta-catenin in vitro. However, the beta-catenin cleavage products generated by caspase-8 were different from those generated by caspase-3 or caspase-6. Caspase-1, -2, -4/11 and -7 did not or only very inefficiently cleave beta-catenin. These data suggest that activation of procaspase-3, -6 or -8 by different stimuli in the cell might result in a differential proteolysis of beta catenin. PMID- 10481057 TI - Blockade of mitochondrial Ca2+ uptake by mitochondrial inhibitors amplifies the glutamate-induced calcium response in cultured cerebellar granule cells. AB - The objective of this study was to evaluate the role of mitochondrial Ca2+ uptake (MCU) in modulation (shaping) of the glutamate (Glu)-induced changes in neuronal cytoplasmic Ca2+ ([Ca2+]i). In order to block MCU, nerve cells were treated with mitochondrial inhibitors (MI) inducing collapse of the mitochondrial potential (Delta Psim). Measurements of changes in [Ca2+]i were performed using either the low-affinity (fura-2FF) or high-affinity (fura-2) Ca2+ indicators. Loading of nerve cells with rhodamine 123 made it possible to monitor changes in Delta Psim. In the first series of experiments it was shown that blockade of MCU in fura-2FF loaded cells with a cocktail of rotenone (2 microM)+oligomycin (2.5 microg/ml) greatly (2.53+/-0.4 times, n=61) increased the [Ca2+]i response to a 1-min Glu (100 microM) pulse. In fura-2-loaded cells, this increase was small (less than 1.3 times) or absent. In the second series of experiments, cocktails of rotenone+oligomycin or FCCP (1 microM)+oligomycin were applied during a prolonged Glu application. This produced strong mitochondrial depolarisation and an additional [Ca2+]i increase. In most cells the latter could be reversed or prevented by a removal of external Ca2+. The MI-induced additional [Ca2+]i increase was especially pronounced in cells loaded with fura-2FF. In some neurones a removal of external Ca2+ did not produce a decrease in [Ca2+]i during combined Glu+MI application, suggesting an impairment of [Ca2+]i extrusion mechanisms of these cells. The conclusion is drawn that MCU makes a considerable contribution to regulation of [Ca2+]i responses caused by Ca2+ influx via Glu activated ionic channels. The reasons for a quantitative difference between [Ca2+]i responses observed in fura-2- and fura-2FF-loaded neurones are discussed. PMID- 10481059 TI - An intronic polymorphic domain often associated with susceptibility to affective disorders has allele dependent differential enhancer activity in embryonic stem cells. AB - Variable number tandem repeats (VNTR) within non-coding regions of a number of genes have been correlated with susceptibility to various disease states. In particular, a VNTR polymorphism of a 16 or 17 bp element within intron 2 of the human serotonin transporter gene has been correlated with a predisposition to affective disorders. We have demonstrated that this region will support differential levels of reporter gene expression in differentiating embryonic stem cells, this being dependent on the presence of 10 or 12 copies of the repeat. The VNTR domain can therefore act as a transcriptional regulator, a property which potentially contributes to disease susceptibility. PMID- 10481060 TI - Amino acid substitution in alpha-helix 7 of Cry1Ac delta-endotoxin of Bacillus thuringiensis leads to enhanced toxicity to Helicoverpa armigera Hubner. AB - Insecticidal proteins or delta-endotoxins of Bacillus thuringiensis are highly toxic to a wide range of agronomically important pests. The toxins are formed of three structural domains. The N-terminal domain is a bundle of eight alpha helices and is implicated in pore formation in insect midgut epithelial membranes. All the delta-endotoxins share a common hydrophobic motif of eight amino acids in alpha-helix 7. A similar motif is also present in fragment B of diphtheria toxin (DT). Site-directed mutagenesis of Cry1Ac delta-endotoxin of B. thuringiensis was carried out to substitute its hydrophobic motif with that of DT fragment B. The mutant toxin was shown to be more toxic to the larvae of Helicoverpa armigera (cotton bollworm) than the wild-type toxin. Voltage clamp analysis with planar lipid bilayers revealed that the mutant toxin opens larger ion channels and induces higher levels of conductance than the wild-type toxin. PMID- 10481061 TI - NMR characterization of the NADP(H)-binding domain of Escherichia coli transhydrogenase: sequential assignment and global fold. AB - The soluble NADP(H)-binding domain of Escherichia coli transhydrogenase (186 amino acids, 20.4 kDa, rotational correlation time 14 ns) was characterized using NMR techniques. The global fold is similar to that of a classical dinucleotide binding fold with six parallel beta-strands in a central sheet surrounded by helices and irregular structures, but is lacking both alphaD and alphaE. The substrate is bound in an extended conformation at the C-terminal end of the parallel beta-sheet and our data support the notion of a redox dependent structural rearrangement. PMID- 10481062 TI - Transient outward K+ currents across the plasma membrane of laticifer from Hevea brasiliensis. AB - Non-inactivating outward rectifying K+ channel currents have been identified in a variety of plant cell types and species. The present study of laticifer protoplasts from Hevea brasiliensis, cells which are specialized for stress response, has revealed, through a switch-clamp method, an outward rectifying current displaying rapid inactivation. The inactivation depended on the external K+ concentration and on the voltage. This current inactivation appeared clearly different from all those previously described in plant cells and it shared homology with current kinetics of animal Shaker family channels. These results, given the recent cloning of plant K+ channel beta-subunits, shed new light on possible plant K+ channel regulation. PMID- 10481063 TI - Cloning of an alpha-glucosidase gene from Thermococcus hydrothermalis by functional complementation of a Saccharomyces cerevisiae mal11 mutant strain. AB - alpha-Glucosidase is found in methanogenic and thermophilic archaea and also in eukaryotes and bacteria. The gene encoding the enzyme was cloned from Thermococcus hydrothermalis by complementation of a Saccharomyces cerevisiae deficiency maltase mutant strain. The gDNA clone isolated encodes an open reading frame corresponding to a protein of 242 amino acids. The protein shows 42% identity to a Pyrococcus horikoshii unknown ORF but no similarities were obtained with polysaccharidase sequences. PMID- 10481064 TI - A 69 bp fragment in the pyrroline-5-carboxylate reductase promoter of Arabidopsis thaliana activates minimal CaMV 35S promoter in a tissue-specific manner. AB - The Arabidopsis thaliana gene that encodes pyrroline-5-carboxylate reductase (At P5R), the last enzyme in proline biosynthesis in A. thaliana, is developmentally regulated and is highly expressed in cells that divide rapidly or undergo changes in osmotic potential. A 69 bp region (P69; -120 to -51) has previously been identified in a 5' deletion analysis of the At-P5R promoter to be necessary for the basal expression. Here, the essential role of P69 for tissue-specific expression of At-P5R is demonstrated by loss- and gain-of-function experiments. PMID- 10481065 TI - Structure of the human VIPR2 gene for vasoactive intestinal peptide receptor type 2. AB - The VPAC(2) (vasoactive intestinal peptide (VIP)(2)) receptor is a seven transmembrane spanning G protein-coupled receptor which responds similarly to VIP and pituitary adenylate cyclase activating polypeptide (PACAP) in stimulating cAMP production. Recently, we reported the localisation of the human VPAC(2) receptor gene (VIPR2) to chromosome 7q36.3 (Mackay, M. et al. (1996) Genomics 37, 345-353). Here, we describe the characterisation of the VIPR2 gene structure and promoter region. The VIPR2 gene is encoded by 13 exons, the initiator codon of the 438 amino acid open reading frame is located in exon 1 and the termination signal and a poly-adenylation signal sequence are located in exon 13. The 5' untranslated region extends 187 bp upstream of the initiator codon and is extremely GC-rich (80%). The poly-adenylation signal is located 2416 bp downstream of the stop codon. Intron sizes range from 68 bp (intron 11) to 45 kb (intron 4) and the human gene spans 117 kb. PMID- 10481066 TI - An early salicylic acid-, pathogen- and elicitor-inducible tobacco glucosyltransferase: role in compartmentalization of phenolics and H2O2 metabolism. AB - Treatment of tobacco cell suspension cultures with a fungal elicitor of defense responses resulted in an early accumulation of the phenylpropanoid glucosyltransferase TOGT, along with the rapid synthesis and secretion of scopolin, the glucoside of scopoletin. Elicitor-triggered extracellular accumulation of the aglycone scopoletin and of free caffeic and ferulic acids could only be revealed in the presence of diphenylene iodonium, an inhibitor of extracellular H2O2 production. Our results strongly support a role for TOGT in the elicitor-stimulated production of transportable phenylpropanoid glucosides, followed by the release of free antioxidant phenolics into the extracellular medium and subsequent H2O2 scavenging. PMID- 10481067 TI - Differential down-regulation of CD95 or CD95L in chronically HIV-infected cells of monocytic or lymphocytic origin: cellular studies and molecular analysis by quantitative competitive RT-PCR. AB - We analysed the expression of CD95/CD95L in two widely used models for studying the cellular effects of chronic infection with human immunodeficiency virus type 1 (HIV-1), i.e. ACH-2 cells, derived from the lymphocytic cell line A301, and U1, derived from monocytic U937 cells. A301 and ACH-2 mounted the same amount of plasma membrane CD95, while U1 had a consistent decrease in CD95 expression. Using different antibodies, we failed to detect the plasma membrane form of its ligand, CD95L, but we could see the intracellular presence of that molecule in A301 cells and, to a lesser extent, in ACH-2 cells, but not in U937 or U1 cells. To confirm the cytofluorimetric data and quantify the expression of CD95L at the RNA level, we developed a quantitative competitive RT-PCR assay. The HUT78 cell line had about 50,000 copies mRNA/1000 cells, three times more after induction with a phorbol ester and ionomycin. ACH-2 expressed about 400- (basal) or 10- (induced) fold less CD95L mRNA than the parental cell line A301; U937 and U1 were below the limit of detection. In cells of lymphoid origin (ACH-2) chronic HIV infection inhibits the expression of CD95L, the phenomenon occurring at the transcriptional level. In cells of monocytic origin (U1) the infection decreases the plasma membrane expression of CD95. This suggests that HIV could trigger different anti-apoptotic strategies which likely depend upon the cell line which is infected. In monocytic cells which act as a viral reservoir, the expression of the molecule whose binding triggers apoptosis decreases, while in lymphoid cells, capable of exerting cytotoxicity, the expression of a molecule which induces apoptosis is reduced. PMID- 10481069 TI - Arabidopsis knockout mutation of ADC2 gene reveals inducibility by osmotic stress. AB - We isolated an Arabidopsis thaliana mutant line carrying an insertion of the En-1 transposable element at the ADC2 locus. The insertion causes a knockout of the arginine decarboxylase 2 gene. We demonstrated that ADC2 is the gene responsible for induction of the polyamine biosynthetic pathway by osmotic stress. No induction of ADC activity by the osmolite sorbitol could be observed in the homozygous mutant, indicating a predominant role of ADC2 in stress response. ADC activity is reduced in the mutant by 44% under non-stressed conditions and the mutant shows no obvious phenotype. This is the first report of a genetically mapped mutation in the polyamine biosynthetic pathway in plants. PMID- 10481068 TI - Overexpression of L-glutamine:D-fructose-6-phosphate amidotransferase provides resistance to methylmercury in Saccharomyces cerevisiae. AB - To identify novel genes that confer resistance to methylmercury (MeHg), a yeast genomic DNA library was transfected into Saccharomyces cerevisiae. Two functional plasmids were isolated from transfected yeast clones D1 and H5 that exhibited resistance to MeHg. The yeast transfected with plasmid isolated from clone H5 was several-fold more resistant than yeast transfected with plasmid from clone D1. Functional characterization of the genomic DNA fragment obtained from clone H5 determined that the GFA1 gene conferred resistance to MeHg. GFA1 was reported to encode L-glutamine:D-fructose-6-phosphate amidotransferase (GFAT) which catalyzes the synthesis of glucosamine-6-phosphate from glutamine and fructose-6-phosphate. Accumulation of mercury in yeast clone W303B/pGFA1, which contains the transfected GFA1 gene, did not differ from that in control yeast clone W303B/pYES2. The W303B/pGFA1 strain did not show resistance to mercuric chloride, zinc chloride, cadmium chloride or copper chloride, suggesting that the resistance acquired by GFA1 gene transfection might be specific to MeHg. This is the first report of a gene involved in MeHg resistance in eukaryotic cells identified by screening a DNA library. PMID- 10481070 TI - The small intestine can both absorb and glucuronidate luminal flavonoids. AB - We have studied the perfusion of the jejunum and ileum in an isolated rat intestine model with flavonoids and hydroxycinnamates and the influence of glycosylation on the subsequent metabolism. Flavone and flavonol glucosides and their corresponding aglycones are glucuronidated during transfer across the rat jejunum and ileum and this glucuronidation occurs without the need for gut microflora. Furthermore, this suggests the presence of glycosidases as well as UDP-glucuronyl transferase in the jejunum. In contrast, quercetin-3-glucoside and rutin are mainly absorbed unmetabolised. The results suggest that the more highly reducing phenolics are absorbed predominantly as glucuronides (96.5%+/-4.6) of the amount absorbed, whereas monophenolic hydroxycinnamates and monophenolic B ring flavonoids are less predisposed to glucuronidation and higher levels of aglycone (88.1%+/-10.1) are detected on absorption through both the jejunum and ileum. PMID- 10481071 TI - Calorie restriction increases Fas/Fas-ligand expression and apoptosis in murine splenic lymphocytes. AB - One-month-old male ICR mice were fed a nutritionally adequate, semipurified diet, either ad libitum (AL) or calorie restricted (CR) (40% less food) for 6 months and were killed to obtain spleens. Flow cytometric analysis revealed increased proportions of both CD4+ and CD8+ T cells in CR-fed mice compared to AL-fed mice. The T cell subsets of CR-fed mice were also found to have higher levels of plasma membrane Fas receptor expression. Similarly, Fas-ligand (Fas-L) expression was higher in anti-CD3-stimulated CD4+ and CD8+ T cells. CR-fed mice also had increased numbers of annexin V-positive CD4+ and CD8+ T cells in stimulated splenic lymphocytes suggesting an increased potential for apoptosis. Fas and Fas L gene expression in splenic lymphocytes, which correlated closely with the observed increased rate of apoptosis, was significantly increased in CR-fed mice compared to AL-fed mice. In conclusion, these results indicate that CR increases the expression of Fas and Fas-L which may contribute to the known beneficial effects of CR such as prolongation of life span by activating chronic physiologically mediated apoptosis. PMID- 10481072 TI - Transgenic replacement of type V adenylyl cyclase identifies a critical mechanism of beta-adrenergic receptor dysfunction in the G alpha q overexpressing mouse. AB - Chronic activation of Gq coupled receptors, or overexpression of G alpha q, in cardiomyocytes results in hypertrophy, enhanced expression of fetal genes, decreased basal and beta-adrenergic receptor (beta AR) stimulated adenylyl cyclase (AC) activities, and depressed cardiac contractility in vivo. Among several abnormalities of the beta AR-Gs-AC pathway that occur in G alpha q overexpressing transgenic mice, we have investigated whether the observed approximately 45% decrease in type V AC expression and function compared to non transgenic (NTG) is the basis of the above phenotype. Transgenic mice were generated that overexpressed by approximately 50% the rat type V AC in the heart using the alpha-myosin heavy chain promoter. These mice were mated with the G alpha q transgenics resulting in animals (ACV/G alpha q) that had restored levels of forskolin stimulated AC activities in cardiac membranes. In addition, basal cardiac AC activities were normalized in the ACV/G alpha q mice (NTG=23+/-4.4, G alpha q=14+/-3.6, ACV/G alpha q=29+/-5.3 pmol/min/mg) as were maximal isoproterenol stimulated activities (59+/-8.9, 34+/-4.6, 52+/-6.7 pmol/min/mg respectively). Cardiac contractility was also improved by ACV replacement, with increased fractional shortening (51+/-2%, 36+/-6%, 46+/-3% respectively). In contrast, hypertrophy and expression of hypertrophy associated fetal genes were not affected. Thus the observed decrease in type V AC that accompanies the development of the cardiac phenotype in the G alpha q model is the dominant mechanism of dysfunctional beta AR signalling and contractility. In contrast, the decrease in type V AC or beta AR signalling to cAMP is not the basis of the hypertrophic response. PMID- 10481073 TI - Binding of the third Na+ ion to the cytoplasmic side of the Na,K-ATPase is electrogenic. AB - A new experimental setup was constructed to allow parallel measurements of total internal reflection fluorescence and of capacitance changes in Na,K-ATPase containing membranes. Effects correlated with cytoplasmic sodium binding to Na,K ATPase were investigated. Ion binding-induced fluorescence changes of the electrochromic dye RH421 in membrane fragments adsorbed on a transparent capacitative electrode corresponded perfectly to capacitance increases detected by a lock-in technique. From these electric measurements it was possible to estimate a dielectric coefficient of about 0.25 for the electrogenic binding of the third Na+ ion. Binding of K+ to cytoplasmic sites was electroneutral. PMID- 10481075 TI - Kinetics of and intermediates in a photocycle branching reaction of the photoactive yellow protein from Ectothiorhodospira halophila. AB - We have studied the kinetics of the blue light-induced branching reaction in the photocycle of photoactive yellow protein (PYP) from Ectothiorhodospira halophila, by nanosecond time-resolved UV/Vis spectroscopy. As compared to the parallel dark recovery reaction of the presumed blue-shifted signaling state pB, the light induced branching reaction showed a 1000-fold higher rate. In addition, a new intermediate was detected in this branching pathway, which, compared to pB, showed a larger extinction coefficient and a blue-shifted absorption maximum. This substantiates the conclusion that isomerization of the chromophore is the rate-controlling step in the thermal photocycle reactions of PYP and implies that absorption of a blue photon leads to cis-->trans isomerization of the 4-hydroxy cinnamyl chromophore of PYP in its pB state. PMID- 10481074 TI - A GSK3-binding peptide from FRAT1 selectively inhibits the GSK3-catalysed phosphorylation of axin and beta-catenin. AB - The Axin-dependent phosphorylation of beta-catenin catalysed by glycogen synthase kinase-3 (GSK3) is inhibited during embryogenesis. This protects beta-catenin against ubiquitin-dependent proteolysis, leading to its accumulation in the nucleus, where it controls the expression of genes important for development. Frequently rearranged in advanced T-cell lymphomas 1 (FRAT1) is a mammalian homologue of a GSK3-binding protein (GBP), which appears to play a key role in the correct establishment of the dorsal-ventral axis in Xenopus laevis. Here, we demonstrate that FRATtide (a peptide corresponding to residues 188-226 of FRAT1) binds to GSK3 and prevents GSK3 from interacting with Axin. FRATtide also blocks the GSK3-catalysed phosphorylation of Axin and beta-catenin, suggesting a potential mechanism by which GBP could trigger axis formation. In contrast, FRATtide does not suppress GSK3 activity towards other substrates, such as glycogen synthase and eIF2B, whose phosphorylation is independent of Axin but dependent on a 'priming' phosphorylation. This may explain how the essential cellular functions of GSK3 can continue, despite the suppression of beta-catenin phosphorylation. PMID- 10481076 TI - Fibre type specificity of haem oxygenase-1 induction in rat skeletal muscle. AB - The expression of the inducible haem oxygenase (HO-1) gene was examined in different skeletal muscles. Rats were treated with haemin and a time course of HO 1 mRNA expression was determined in soleus and extensor digitorum longus (EDL) muscles. Fibre type composition and tissue myoglobin content were also measured. We found that HO-1 mRNA expression markedly increased in soleus (type I fibres) muscle but was only slightly affected in EDL (type II fibres). HO-1 expression directly correlated with both percentage of red fibres and tissue myoglobin. These data demonstrate that HO-1 gene expression follows a fibre type-specific pattern which might indicate an important role for this protein in the maintenance of skeletal muscle function. PMID- 10481077 TI - Effects of acute leptin administration on the differences in proton leak rate in liver mitochondria from ob/ob mice compared to lean controls. AB - In this investigation, the effects on proton leak of leptin administration to ob/ob mice was measured for liver mitochondria. We and others have shown that proton leak is approximately 3 times greater in liver mitochondria from ob/ob mice compared to lean controls at any given membrane potential. The results are consistent with obese mammals having higher lean mass-specific metabolic rates compared to lean controls. The increase in proton leak rate at any given membrane potential cannot be explained by the presence of free fatty acids associated with mitochondria isolated from the obese animals. The difference in proton leak must therefore represent a real difference in inner membrane permeability. Acute leptin (OB protein) administration restores the liver mitochondrial proton leak rate of ob/ob mice to that of lean controls. There was no effect on proton leak rate of liver mitochondria from sham-treated ob/ob mice. These novel results indicate a role for leptin, either directly or indirectly, in regulating the efficiency of oxidative phosphorylation. PMID- 10481078 TI - Identification and characterization of splice variants of ephrin-A3 and ephrin A5. AB - Ephrins and Eph receptors have been implicated to play important roles in axon guidance. A variable spacer region exists that differs significantly among distinct ephrins. An ephrin-A5 isoform has previously been isolated which lacks 27 amino acids within the spacer region. The expression and biological activities of this isoform, as well as the existence of isoforms for other ephrins that show variation within the spacer region, remain unknown. We report here a novel alternatively spliced isoform of ephrin-A3 which lacks the corresponding variable region. When compared to the longer isoforms, the shorter isoforms of both ephrin A3 and ephrin-A5 remained less prominent in the brain during development, though their expression increased at postnatal stages. In addition, they could inhibit neurite outgrowth of dorsal root ganglia (DRG) neurons, suggesting that the corresponding variable regions were not essential for their axon guidance activities. PMID- 10481079 TI - Regulation of bacterial photosynthesis genes by oxygen and light. AB - Most bacteria have the capability to adapt to changes in their environment. Facultatively phototrophic bacteria like Rhodobacter can switch from aerobic respiration to anoxygenic photosynthesis in the absence of oxygen. The formation of the photosynthetic apparatus is primarily regulated by oxygen tension. The amount of photosynthetic complexes is influenced by the light intensity in anaerobic cultures. This review focuses on the molecular mechanisms involved in the regulation of Rhodobacter photosynthesis genes by oxygen and light. PMID- 10481081 TI - Mapping studies and genetic analysis of transfer genes of the multi-resistant IncHI2 plasmid, R478. AB - A bank of transfer-defective Tn7 insertion mutants of the multi-resistant IncHI2 megaplasmid, R478, was generated. Complementation analysis of these mutations identified a large 144-kb transfer-associated region of R478. A 6.8-kb segment from the transfer region was sequenced. The precise locations of Tn7 insertion within four distinct R478::Tn7 transfer-defective mutants were mapped and each insertion was found to disrupt a specific open reading frame. These transfer associated determinants of R478 were designated htdB (H transfer determinant), htdD, htdT and htdC. Both htdB and htdC encoded amino acid sequences that showed a low homology with pilus biosynthetic proteins encoded by the F plasmid. PMID- 10481080 TI - Identification of a virulence-associated protein homolog gene and ISRa1 in a plasmid of Riemerella anatipestifer. AB - Riemerella anatipestifer is the causative agent of polyserositis of ducks and geese. We have previously reported that a 3.9-kb plasmid, pCFC1, carries protein genes (vapD1 and vapD2) that are similar to virulence-associated genes of other bacteria. In the present study, we report the complete sequence of a second plasmid of 5.6 kb, pCFC2. pCFC2 has a 28% G-C content and three large open reading frames (ORFs). One of the ORFs (designated asVapD1) encodes a polypeptide that shares 53.9, 53.9, 48.3, 48.3 and 46.1% identity with virulence-associated proteins of Dichelobacter nodosus, Actinobacillus actinomycetemcomitans, Neisseria gonorrhoeae, Helicobacter pylori and Haemophilus influenzae, respectively. The second ORF encodes a putative DNA replication protein (RepA3) with 309 amino acids and a molecular mass of approximately 36 kDa. A novel insertion sequence (IS) element, designated ISRa1, was found on the plasmid pCFC2. ISRa1 was flanked by 15-bp imperfect inverted repeats (only one mismatched nucleotide). ISRa1 contained an ORF encoding a putative transposase of 292 amino acids. Southern blot analysis indicated that in R. anatipestifer strains examined, ISRa1 was present with 2-20 copies (at least). ISRa1 displayed a sequence approximately 35% homologous to the putative IS982 and RSBst-alpha from Lactococcus lactis ssp. cremoris SK11 and Bacillus stearothermophilus CU21. Three hybridization patterns of genomic DNA of eight R. anatipestifer strains with an ISRa1 probe indicated that ISRa1 might be a useful tool for epidemiological studies. PMID- 10481082 TI - Molecular characterization of a flagellar (fla) operon in the oral spirochete Treponema denticola ATCC 35405. AB - A Treponema denticola 9.6-kb motility locus containing 11 genes was identified, sequenced and analyzed. The genes were designated tap1, flgD, flgE, orf4, motA, motB, fliL, fliM, fliY, orf10 and fliP. The order of these genes is identical to that of the corresponding region of the Treponema pallidum fla operon. Seven of the deduced Fla proteins share significant homology with both Escherichia coli and Bacillus subtilis proteins associated with flagellar structure and function. Reverse transcription-PCR analysis indicated that the T. denticola fla genes are transcribed as a single unit. A putative sigma(28)-like promoter, virtually identical to the T. pallidum fla promoter, was identified upstream of tap1. These results showed that the T. denticola and T. pallidum fla operons are highly conserved, supporting the proposed phylogenetic relatedness of these spirochetes. PMID- 10481083 TI - A bifunctional transposon mini-Tn5gfp-km which can be used to select for promoter fusions and report gene expression levels in Agrobacterium tumefaciens. AB - A mini-Tn5 transposon derivative, mini-Tn5gfp-km, has been constructed which contained a promoter-less artificial operon consisting of two open reading frames, green fluorescent protein (GFP) and neomycin phosphotransferase II (NptII). When this transposon was used to mutagenize Agrobacterium tumefaciens, all the mutants selected in the presence of kanamycin exhibited GFP expression, which could be conveniently monitored by a fluorometer. The transposon appeared to be bifunctional and could provide both selection and reporter functions. Even the mutants showing minimal levels of GFP expression were still resistant to kanamycin. This suggests that this transposon can be used to select for insertions downstream of both weak and strong promoters, as long as the insertions themselves are non-lethal. This system was used to identify A. tumefaciens genes that were upregulated in response to acidic pH. Screening only 20 colonies led to identification of two promoters that were specifically induced by low pH and one promoter that was specifically induced by acetosyringone in a minimal medium of pH 5.5. PMID- 10481084 TI - Anionic polymers of Bacillus subtilis cell wall modulate the folding rate of secreted proteins. AB - In order to characterize the dynamics of the interaction between the emergent membrane translocated exoprotein and the components of Bacillus subtilis cell wall, we examined the kinetics of the in vitro refolding of levansucrase and alpha-amylase, at pH 7 and 37 degrees C, in the presence of polyphosphates (polyP) of various chain lengths (2/=16. In contrast, polyP modulate indirectly the rate of alpha-amylase refolding via their affinity for calcium. These differential effects might explain that the rate of the cell wall translocation of alpha-amylase secretion was found to be half that of levansucrase. PMID- 10481085 TI - Genetic variability in Metarhizium flavoviride revealed by telomeric fingerprinting. AB - Previous studies using arbitrarily primed PCR (AP-PCR/RAPD) analysis have shown only little genetic variation among isolates of the entomopathogenic fungus Metarhizium flavoviride. In the current study, however, telomeric fingerprinting unambiguously differentiated several Brazilian strains of M. flavoviride as well as strains from Africa and Australia. Using this technique, similarity estimates of telomeric DNA among distinct strains were less than 50%, showing this locus to be highly mutable in this species. PMID- 10481086 TI - A transposon insertion in the gumG homologue of Xanthomonas oryzae pv. oryzae causes loss of extracellular polysaccharide production and virulence. AB - Xanthomonas oryzae pv. oryzae causes a serious disease of rice called bacterial leaf blight. It produces copious amounts of extracellular polysaccharide (EPS). An EPS- and virulence-deficient mutant of X. oryzae pv. oryzae was isolated by Tn5 mutagenesis. The mutant allele in this strain was cloned by transposon tagging in the Escherichia coli vector pBluescript and the DNA sequences flanking the transposon insertion site were determined. Computer-based similarity searches in the DNA database using the BLAST algorithm showed these sequences to be 78% identical at the nucleotide level to a gene, gumG, in the gum cluster, which is required for EPS biosynthesis in Xanthomonas campestris pv. campestris. A 36-kb X. oryzae pv. oryzae genomic clone containing the putative EPS biosynthetic gene cluster of X. oryzae pv. oryzae restored both EPS production and virulence proficiency to the gumGXo::Tn5 mutant. The results suggest that EPS is an important virulence factor of X. oryzae pv. oryzae. PMID- 10481087 TI - Purification of a biologically active recombinant glyceraldehyde 3-phosphate dehydrogenase from Candida albicans. AB - We report here the purification of a functionally active recombinant glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Candida albicans. The GAPDH protein encoded by the TDH1 gene was obtained as a glutathione S-transferase fusion protein by expression in the vector pGEX-4T-3, and purified by affinity chromatography and thrombin digestion. The purified protein displays GAPDH enzymatic activity (42 micromol NADH min(-1) mg(-1)) as well as the laminin and fibronectin binding activities previously described. In addition, the recombinant GAPDH is covalently modified by NAD linkage; this modification is stimulated by nitric oxide and probably involves a sulfhydryl group (cysteine) residue since it is inhibited by Hg(2+) and cysteine. PMID- 10481088 TI - nalB-type mutations causing the overexpression of the MexAB-OprM efflux pump are located in the mexR gene of the Pseudomonas aeruginosa chromosome. AB - Two mutations, one at the nalB and the other at the mexR locus, in the Pseudomonas aeruginosa chromosome are known to cause overexpression of the MexAB OprM efflux pump. Based on the following results, we concluded that nalB is the mutation that has occurred within the mexR gene of the P. aeruginosa chromosome. (i) Nucleotide sequencing of the mex operon upstream region of 21 independent nalB-type mutants including the original nalB9 revealed that all the mutations were located within the mexR gene. The mutations were classified into three different groups and nine types including single base substitutions, single base deletions and base insertions. (ii) Substitution of the mutant mexR with the wild type mexR and replacement of the wild-type mexR with a defined mexR mutation resulted in the expression of wild-type and nalB-type MexAB-OprM respectively, which was confirmed by testing the antibiotic susceptibility and beta galactosidase activity of the mexA-lacZ translational fusion. PMID- 10481089 TI - Apoptosis of murine peritoneal macrophages induced by an avian pathogenic strain of Escherichia coli. AB - The mechanisms used by avian strains of Escherichia coli to invade the respiratory epithelia, leading to septicemia in poultry, are not well established. In this work, we show that resident murine peritoneal macrophages infected in vitro with an avian strain of E. coli underwent apoptosis 4 h after infection (55.6% of apoptosis in infected cells versus 3.5% in non-infected cells). Heat-inactivated bacteria did not induce apoptosis and the inhibition of phagocytosis by pretreatment of cells with cytochalasin D reduced the number of apoptotic cells from 55.6 to 13.9% (P<0.05), showing that the bacteria must be intracellularly located and viable to induce apoptosis. Therefore, these data suggest that induction of macrophage apoptosis may be a pathogenic mechanism employed by avian E. coli to circumvent the host defences and invade the respiratory epithelia. PMID- 10481090 TI - Characterization of the gltF gene product of Escherichia coli. AB - The glt operon of Escherichia coli comprises the structural genes for the glutamate synthase subunits (gltB and gltD) and gltF, whose product was previously suggested to have regulatory functions. The A/T-rich region between gltD and gltF contains a weak promoter and a translation initiation site for gltF. The GltF protein is preceded by a signal peptide, which is cleaved off during export into the periplasmic space. A gltF::Km(R) insertion mutant was constructed and shown here to have no detectable phenotype with respect to amino acid utilization or ammonium transport. Thus, GltF is apparently not involved in regulation of nitrogen catabolism. PMID- 10481092 TI - Identification of cytochrome c oxidase in the alkaliphilic, obligately chemolithoautotrophic, sulfur-oxidizing bacterium 'Thioalcalomicrobium aerophilum' strain AL 3. AB - Cytochrome c oxidase from the novel alkaliphilic autotrophic sulfur bacterium 'Thioalcalomicrobium aerophilum' strain AL 3 was isolated and purified 87-fold. Spectroscopic analysis revealed the presence of both c- and b-type hemes as well as copper in a ratio of 3:2:1. The purified enzyme consists of three subunits with apparent molecular masses of 41, 34 and 32 kDa. The two small subunits contain covalently bound heme c. With TMPD as a substrate the pH optimum was determined to be pH 8.0. In the presence of monovalent cations the specific activity of the purified oxidase increased significantly. The enzyme was not able to oxidize external cytochrome c, but accepted electron from its native electron donor. The latter was separated from the other membrane cytochromes during anion exchange chromatography and was identified as a high potential cytochrome c(551). Overall the data indicate that the cytochrome c oxidase from this alkaliphilic autotrophic bacterium belongs to the heme-copper oxidase superfamily; regarding its subunit composition and content of prosthetic groups, the enzyme is similar in many aspects to the cbb(3)-type cytochrome c oxidases described for several neutrophilic bacteria, including anaerobic phototrophic and aerobic sulfur oxidizing bacteria. PMID- 10481091 TI - The Pseudomonas aeruginosa algC gene product participates in rhamnolipid biosynthesis. AB - Pseudomonas aeruginosa produces exoproducts correlated with its pathogenicity. One of these virulence-associated traits is the surfactant rhamnolipid. The production of alginate and lipopolysaccharide (LPS) are also of importance for P. aeruginosa virulence. The product of the algC gene (which is involved in alginate production through its phosphomannomutase activity and in LPS synthesis through its phosphoglucomutase activity) participates in rhamnolipid production, presumably catalyzing the first step in the deoxy-thymidine-diphospho-L-rhamnose (dTDP-L-rhamnose) pathway, the conversion of glucose-6-phosphate to glucose-1 phosphate. Other structural alg genes, encoded in the alg operon, are not involved in rhamnolipid nor LPS production. These results show that the AlgC protein plays a central role in the production of the three P. aeruginosa virulence-associated saccharides: alginate, LPS and rhamnolipid. PMID- 10481093 TI - Controlled specific expression and purification of 6 x His-tagged proteins in Pseudomonas. AB - The 6 x His affinity tags have proved invaluable for the exclusive purification of proteins, in Escherichia coli, of genes cloned in frame with a 6 x His tag and a strong inducible promoter. Here, we demonstrate that the system can be extended to Gram-negative bacteria other than E. coli, by the use of compatible broad-host range plasmids. As an example, the inducible synthesis and specific purification of the Pseudomonas 6 x His-PfrA siderophore regulatory protein in Pseudomonas putida WCS358 is demonstrated. PMID- 10481094 TI - By-product formation during exposure of respiring Saccharomyces cerevisiae cultures to excess glucose is not caused by a limited capacity of pyruvate carboxylase. AB - Upon exposure to excess glucose, respiring cultures of Saccharomyces cerevisiae produce substantial amounts of ethanol and acetate. A possible role of a limited anaplerotic capacity in this process was investigated by overexpressing pyruvate carboxylase and by replacing it with a heterologous enzyme (Escherichia coli phosphoenolpyruvate carboxylase). Compared to the wild-type, neither the pyruvate carboxylase (Pyc)-overexpressing nor the transgenic strain exhibited reduced by product formation after glucose pulses to aerobic glucose-limited chemostat cultures. An increased intracellular malate concentration was observed in the two engineered strains. It is concluded that by-product formation in S. cerevisiae is not caused by a limited anaplerotic capacity. PMID- 10481095 TI - Distribution of Vibrio cholerae O1 antigen biosynthesis genes among O139 and other non-O1 serogroups of Vibrio cholerae. AB - The organization and distribution of the genes responsible for O antigen biosynthesis in various serogroups of Vibrio cholerae were investigated using several DNA probes derived from various regions of the genes responsible for O1 antigen biosynthesis. Based on the reactivity pattern of the probes against the various serogroups, the cluster of genes responsible for the O1 antigen biosynthesis could be broadly divided into six groups, designated as class 1-6. The class 3 cluster of genes corresponding to gmd to wbeO, wbeT and a part of wbeU was specific for only the O1 serogroup. The other cluster of genes (class 1, 2, 4-6) reacted with other serogroups of V. cholerae. These data indicate that serotype conversion in V. cholerae does not depend on a simple mutational event but may involve horizontal gene transfer not only between V. cholerae strains but also between V. cholerae and species other than V. cholerae. PMID- 10481096 TI - Inability of encapsulated Klebsiella pneumoniae to assemble functional type 1 fimbriae on their surface. AB - We screened phase variants of Klebsiella pneumoniae isolates for the expression of capsule and type 1 fimbriae and found that all of the 22 blood isolates were encapsulated and did not express type 1 fimbriae while 10 of 11 urinary tract isolates expressed type 1 fimbriae but were unencapsulated. Phase variants from selected isolates were found to be either unencapsulated and fimbriated or lacked both structures. Variants expressing both structures were not detected. Fimbrial subunits FimH and FimA were localized in the periplasmic space of the parent strain and on the surface of the unencapsulated variants. The results suggest that capsule formation impedes assembly of pre-formed fimbrial subunits on the bacterial surface. PMID- 10481097 TI - Engineering of a Staphylococcus carnosus surface display system by substitution or deletion of a Staphylococcus hyicus lipase propeptide. AB - Surface display of recombinant proteins on bacteria and phages has become an important topic in bioscience. A system for the display of heterologous proteins on the surface of Staphylococcus carnosus employs the secretion signal and propeptide from a Staphylococcus hyicus lipase for translocation and since the propeptide is of considerable size (207 amino acids) and not processed in S. carnosus, we have investigated the possibility to delete or substitute the propeptide for smaller protein domains, to thereby improve the surface display system. A set of new vectors was constructed and the surface expression of model proteins was investigated by various methods, including fluorescence-activated cell sorting. The results suggest that the propeptide region indeed can be deleted when proteins which are easily secretable are displayed. In contrast, the propeptide seems to be advantageous for translocation of inefficiently secreted proteins. Moreover, our study also presents a rational strategy for how to monitor the engineering efforts for the optimization of a surface display system. PMID- 10481098 TI - GFP-aided confocal laser scanning microscopy can monitor Agrobacterium tumefaciens cell morphology and gene expression associated with infection. AB - We tagged Agrobacterium tumefaciens cells with a mini-Tn5 transposon containing a promoterless gene encoding a green fluorescent protein (GFP). Some of the GFP tagged individual bacterial cells exhibited strong green fluorescence, which reflected the expression levels of the GFP-tagged genes. Those cells could be readily detected with a confocal laser scanning microscope (CLSM). We observed that the fluorescence and morphology of A. tumefaciens cells grown in plant tissues resembled those grown in a minimal medium of low pH, which is required for expression of the virulence genes responsible for tumorigenesis. This suggests that GFP-aided CLSM can be used to determine which growth medium is more representative of the nutritional conditions that a pathogen encounters in plant tissues. We also observed that the fluorescence and morphology of A. tumefaciens cells changed dramatically during the course of infection. Our data suggested that A. tumefaciens cells were probably better fed upon successful colonization. We believe that GFP-aided CLSM can help study the fate of A. tumefaciens cells inside plant tissues by monitoring cell morphology and gene expression associated with the infection process in situ. PMID- 10481099 TI - Purification and characterization of a novel enzyme, L-threo-3-hydroxyaspartate dehydratase, from Pseudomonas sp. T62. AB - L-threo-3-Hydroxyaspartate dehydratase (L-threo-3-hydroxyaspartate hydro-lyase), which exhibited specificity for L-threo-3-hydroxyaspartate (K(m)=0.74 mM, V(max)=37.5 micromol min(-1) (mg protein)(-1)) but not for D-threo or D, L erythro-3-hydroxyaspartate, was purified from a cell-free extract of Pseudomonas sp. T62. The activity of the enzyme was inhibited by hydroxylamine and EDTA, which suggests that pyridoxal 5'-phosphate and divalent cations participate in the enzyme reaction. The NH(2)-terminal amino acid sequence showed significant similarity to the Saccharomyces cerevisiae YKL218c gene product, a hypothetical threonine dehydratase. However, the purified enzyme showed no threonine dehydratase activity. PMID- 10481100 TI - A single-step transconjugation system for the introduction of unmarked deletions into Actinobacillus pleuropneumoniae serotype 7 using a sucrose sensitivity marker. AB - Research on the porcine respiratory tract pathogen Actinobacillus pleuropneumoniae requires the availability of improved genetic tools. Therefore, using the sacB gene of Bacillus subtilis, we developed a sucrose-based counterselection system that allows rapid curing of an Escherichia coli-A. pleuropneumoniae shuttle vector as well as the introduction of unmarked mutations into the A. pleuropneumoniae chromosome. A cassette containing the Tn903 kanamycin resistance determinant (km(r)) and the sacB gene expressed from the A. pleuropneumoniae omlA promoter was introduced by homologous recombination into the ureC gene of A. pleuropneumoniae. The resultant stable plasmid cointegrates were kanamycin-resistant, sucrose-sensitive, and urease-positive. A simple counterselection on sucrose-containing agar plates without an additional transconjugation step allowed the efficient isolation of urease-negative A. pleuropneumoniae mutants that had lost the km(r)-sacB cassette. PMID- 10481101 TI - The leukotoxin of Pasteurella haemolytica binds to beta(2) integrins on bovine leukocytes. AB - The putative receptor proteins of Pasteurella haemolytica leukotoxin were isolated from bovine polymorphonuclear neutrophil lysate by affinity chromatography on a leukotoxin-specific monoclonal antibody column to which the leukotoxin was pre-bound. SDS-PAGE of the purified proteins showed four bands at 180 kDa, 170 kDa, 150 kDa and 95 kDa, in addition to the expected 102-kDa leukotoxin band and a series of bands with molecular masses lower than 102 kDa representing the disintegrated leukotoxin. N-terminal amino acid sequencing of the 170-kDa band showed homology with human and murine CD11b. The purified proteins reacted specifically with monoclonal antibodies specific for CD11a, CD11b, CD11c (the alpha chains of beta(2) integrins), and CD18 (the beta chain of beta(2) integrins). Pre-incubation of polymorphonuclear neutrophils with a monoclonal antibody specific for CD18 reduced the cytotoxicity of the leukotoxin to the cells. These results indicate that the leukotoxin binds to the beta(2) integrins on bovine leukocytes, very likely via CD18. PMID- 10481102 TI - Lack of expression of bundle-forming pili in some clinical isolates of enteropathogenic Escherichia coli (EPEC) is due to a conserved large deletion in the bfp operon. AB - Enteropathogenic Escherichia coli (EPEC) produces a plasmid-encoded type IV pilus, called the bundle-forming pilus (BFP), involved in the formation of the localized adhesion onto epithelial cells. In this study, we demonstrate that clinical isolates of serotypes O128ab:H2 and O119:H2 contain a ca. 13-kb deletion in the bfp operon, resulting in a lack of expression of these pili. An IS sequence with homology to the IS66 of Agrobacterium tumefaciens replaced the deleted bfp genes. These results suggest that the bfp operon was deleted through a transpositional event and that other adherence factors may mediate attachment of these bacteria to the host cells. PMID- 10481105 TI - Corrigendum to "Investigation of steroid receptor function in the budding yeast Saccharomyces cerevisiae". PMID- 10481103 TI - Type II restriction endonucleases from Helicobacter pylori include an enzyme with a novel recognition sequence. AB - Type II restriction endonuclease activities of Helicobacter pylori strain Roberts and of the type strain H. pylori NCTC 11637 were detected and analysed by conventional techniques. The endonucleases were partially purified, their optima for activity and their recognition and cleavage sites were determined. H. pylori (Roberts) contained at least two enzymes: HpyBI was an isoschizomer of RsaI (GT/AC) and HpyBII was of a novel specificity (GTN/NAC). H. pylori NCTC 11637 was found to contain an isoschizomer of EcoRV (HpyCI: GAT/ATC) and at least one other enzyme which was too unstable to characterise. PMID- 10481106 TI - [Correlations between brain SPECT and neuropsychology assessments in mild and moderate stages of Alzheimer's disease]. AB - Thirty-four patients with a probable Alzheimer's Disease (AD) (mild AD = 16; moderate AD = 18) and 12 matched controls were evaluated using semiquantitative SPECT-99mTc-HMPAO and neuropsychological tests (CERAD). RESULTS: Both temporal hypoperfusion (p < 0.01) and memory tests (p < 0.001) made it possible to differentiate the controls from mild AD patients. In these patients, significant correlations (p < 0.05) were also found between: 1) delayed recall test/temporal hypoperfusion, 2) learning memory test/temporoparietal and frontal hypoperfusion and 3) visual constructive praxis/posterior temporal hypoperfusion. In contrast to mild AD, moderate AD patients showed higher temporal (p < 0.01) parietal and frontal (p < 0.05) hypoperfusion along with worsening of praxis (p < 0.001) and memory tests (p < 0.05). CONCLUSIONS: SPECT imaging and neuropsychology evaluation can distinguish controls from AD patients with mild an moderate grades of dementia, showing a strong correlation from the early stages of AD. PMID- 10481107 TI - [Use of a cyclotron in the production of positron emitting radionuclides]. AB - The experience acquired by our center during the first two years of using cyclotron 18/9 (IBA) dedicated to the production of clinical positron emission radionuclides is described. The cyclotron performance characteristics, production yields, quality control and synthesized radiotracers are analyzed. Cyclotron makes it possible to produce up to 3,300 mCi of 18F-, 270 mCi of 18F2, 3,100 mCi of 11C, 502 mCi of 13N (in 120, 60, 35 and 20 minutes respectively) and 540 mCi/min of 15O. In our center, about 85% of the PET studies are performed with 18F-FDG, whereas the remaining are done with 15O-water, 11C-bicarbonate, 11C methionine, 13N-ammonia or 18F-. Cyclotron is included in the Radiopharmacy Unit of our PET facility and is subjected to a global quality control program. Follow up of the bombardment parameters and periodic verifications of the cyclotron performance have made it possible to prevent equipment functioning problems, increase mean time between stoppage and decrease downtime. We conclude that cyclotron has high production capabilities and allows enough flexibility for a clinical and research positron emission tomography center; furthermore, it can also be used for regional distribution of 18F-FDG to satellite PET centers. PMID- 10481108 TI - [Plasma prostaglandin E2 in pregnant women undergoing labor induction with endocervical gel application]. AB - The authors describe an assay that quantify with precision the plasmatic prostaglandin E2 (PGE2). The aim of this study was to evaluate the effect of the gel formulation PGE2 endocervical in the induction of labour. We have studied 26 pregnant women without disease at the term of the pregnancy. The blood sample was collected before and 30 minutes after the labor induction in tubes with 10 mg EDTA Na2 and 5 microg of aspirin. The PGE2 was isolated by extraction and chromatography with 70-100% recovery. The concentration was evaluated by a completion assay with RIA. The PGE2 in plasma was found before and after the induction of labor with the following results: 347 +/- 51.6 and 719 +/- 128 pg/ml. The data was compared by the Student's t test for paired values (df = 50, t = 13. 22 and p < 0.0001). We can conclude that in pregnant women the concentration of PGE2 in plasma was double after the labor induction with the sensitivity in 1.25 pg/tube and the reproducibility 4.6% intraassay and 7% interassay showing that the method is sensitive, reproducible and efficient for the PGE2. PMID- 10481109 TI - [Validation of the preparation of individual doses of 51Cr-EDTA in syringes]. AB - The 51Cr-EDTA is one of the radiopharmaceuticals more used in the glomerular filtration rate determination. The necessity to prepare a daily reference dose every time that are prepared the patient's doses can go to a poor exploitation of the multidose vial. One way of improving its use consists on the preparation of individual doses in ready syringes to inject, for several days, employing only one of them as a standard. The aim of this study was to evaluate the storing of 51Cr-EDTA individual doses at room temperature and at 4 degrees C during 4 weeks, analyzing the radiochemical purity, the sterility and the radiopharmaceutical retention into the syringe. The results obtained showed that the radiochemical purity remains stable, always been greater than the value recommended for its use (> 95%). Likewise, there were not microorganism contamination nor 51Cr-EDTA appreciable retention into the syringes. We concluded that the preparation of individual doses of 51Cr-EDTA in syringes and their conservation at room temperature, or at 4 degrees C, during a period of 4 weeks, neither influence in the radiopharmaceutical quality nor increase its retention into the syringe. PMID- 10481110 TI - [Quantification of liver volume by SPECT]. AB - This study aimed to optimize a method to estimate liver volume by SPECT. Several phantoms were used to simulate the liver shape with a volume between 400 and 2,500 cc. The SPECT studies were acquired using 360 degrees and 180 degrees circular orbits, 64 projections, 64 x 64 word matrix, all purpose parallel-hole collimator and were reconstructed with a HANN filter with frequency cutoff = 0.5 cycles/ pixel. The reconstructed studies were processed with a program that estimates the organ volume for different threshold cutoff levels for counting. The optimum cutoff threshold level to quantify the liver volume was 50% and 48% for 360 degrees orbits and 180 degrees orbits, respectively. A linear relationship was observed between the real and calculated volumes with r = 0.994 for the 360% orbits and r = 0.976 for the 180 degrees orbits, The average error between corrected and real volumes was 4.2% and 10.87% for both orbits, respectively. The method was applied to 14 patients with different degrees of hepatomegaly who had undergone a liver 99mTc-sulfur colloid scintigraphy. The intra and interobserver variability as well as the influence of the degree of contour adjustment of the phantom mask were studied. No significant differences were observed in the results, so that it can be considered that neither the mask trace nor the operator influence the results. PMID- 10481111 TI - [CEA, CA 19.9 and CA 195 in patients with colorectal carcinoma. ROC analysis]. AB - The aim of this study was to assess the utility of determining serum levels of the CEA, CA 19.9 and CA 195 tumor markers in 906 patients with colorectal carcinoma for a 5-year follow-up period. In addition to calculating sensitivity and specificity for the common normal limits, the results were analyzed with RO curves. Sensitivity values for 90% and 95% specificity were also obtained. The results of the areas under the curve (CEA: 0.887; CA 19.9: 0.802; CA 195: 0.971) and sensitivities and specificities at different normality limits showed better behavior for CEA with respect to CA 19.9 and CA 195, the last two presenting similar results. CEA continues to be the best tumor marker to monitor patients with colorectal carcinoma. PMID- 10481112 TI - [Merkel cell carcinoma. Utility of scintigraphy with 111In-DTPA-pentetreotide]. AB - Merkel cell carcinoma (MCC) is an unusual malignant primary skin tumor, having a high incidence of local recurrent, and regional and distant metastasis. Due to its capacity to express somatostatin receptors, it can be detected in vivo with 111In-pentetreotide scintigraphy (Ostreoscan). We present a case of a MCC whose scintigraphy revealed regional metastases of a primary frontal cutaneous tumor that had been removed previously. The results verified a good correlation with clinical, radiological an histopathological findings. PMID- 10481113 TI - [Bone metastasis versus insufficiency fractures due to pelvic radiotherapy for gynecologic neoplasm]. AB - Five cases of patients with gynecological neoplasm (four cervix carcinoma and one endometrial sarcoma) who underwent pelvic external radiotherapy and intracavitary brachytherapy in whom pathologic pelvic uptake was found in the bone scan are presented. The diagnosis was pelvic insufficiency fractures due to radiotherapy adverse effects on the skeletal system confirmed by CT and by the favorable scintigraphy and clinical outcome. Both bone metastases and insufficiency fractures must be considered in the differential diagnosis of bone pain in irradiated pelvises. The bone scintigraphy detects these insufficiency fractures early and can show a typical symmetric uptake pattern. In asymmetric lesions, the CT and clinical follow-up as well as the scintigraphic evolution of the lesions should confirm the findings of the bone scintigraphies. PMID- 10481114 TI - [Eight year history of neuroblastoma in an adult patient. Value of 123I-MIBG and 111In-DTPA-D-Phe-octreotide scintigraphy]. AB - A case of a 30 year old male with an eight year history of neuroblastoma and whose general health was good is presented. After his last check-up, which included a CT scan and 99mTc-MDP bone scintigraphy, a 123I-MIBG and 111In-DTPA-D Pheoctreotide scintigraphy was performed and provided us with complementary data that contributed to a more precise diagnosis of the location and extension of the neuroblastoma and to the biological features of the tumor. Thus, this report deals with an adult neuroblastoma patient whose general health is good in whom the exact extension of the lesion was determined by a combination of diagnostic imaging techniques. PMID- 10481115 TI - Economic evaluation of the use of octreotide for prevention of complications following pancreatic resection. AB - Recent studies have concluded that octreotide can prevent complications in patients undergoing pancreatic resections. Given the acquisition cost of octreotide, a cost-effectiveness analysis was performed to establish whether if the additional cost associated with its use was justified by a decrease in the consumption of other resources. To evaluate success rates and complication rates, a meta-analysis of double-blind, randomized, controlled clinical trials was conducted. The rates for pancreatic fistula and fluid collection were 10.7% (95% confidence interval [CI] 7.9 to 13.4) and 3.6% (95% CI 1.9 to 5.2) for octreotide vs. 23.4% (95% CI 19.7 to 27. 1) and 8.8% (95% CI 6.2 to 11.3) for placebo. In a second phase we evaluated the treatment cost for patients with and without complications using two different models of cost savings. In the first model the cost to treat a pancreatic fistula was calculated as the per diem rate (as determined by Statistics Canada) multiplied by the incremental length of stay associated with the complication. In the second model we used data from institutions participating in the Ontario Case Costing Project. In model 1 the estimated incremental length of hospital stay attributed to a pancreatic fistula was 7 days, based on a review of the literature, and the per diem was $552. In model 2 the average cost of care for patients with or without complication was $32,347 (n = 17; 95% CI $20,882 to $43,812) and $11, 169 (n = 18; 95% CI $7558 to $14,779), respectively. The data suggest that when compared to placebo, octreotide is a dominant treatment strategy. In model 1, in a cohort of 100 patients, octreotide saved an average of $853 per patient while allowing 16 incremental patients to avoid complications. In model 2 use of octreotide resulted in an average savings of $1642 per patient while still allowing 16 patients to avoid complications. Detailed one-way and two-way sensitivity analyses suggest that both models were robust. The use of octreotide is a cost effective strategy in patients undergoing elective pancreatic resection. Consideration should be given to extending its use to patients who are at high risk for development of complications following pancreatic surgery and who do not have any contraindications to the use of this drug. PMID- 10481116 TI - Major vascular resection as part of pancreaticoduodenectomy for cancer: radiologic, intraoperative, and pathologic analysis. AB - Intraoperative assessment is inaccurate in defining the relationship of a pancreatic head neoplasm to adjacent vascular structures. We evaluated the ability of preoperative contrast-enhanced CT to predict the need for vascular resection during pancreaticoduodenectomy and examined the resected vessels for histologic evidence of tumor invasion. During a 7-year period, 63 patients underwent pancreaticoduodenectomy with en bloc resection of adjacent vascular structures for a presumed pancreatic head malignancy. Clinical, radiologic, operative, and pathologic data were reviewed and analyzed. Fifty-six patients underwent resection of the superior mesenteric-portal vein confluence, three patients required inferior vena cava resection, and the hepatic artery was resected and reconstructed in eight patients. The operative mortality rate was 1.6%, and the overall complication rate was 22%. CT predicted the need for resection of the superior mesenteric or portal veins in 84% of patients. Pathologic analysis revealed tumor invasion of the vein wall in 71% of resected specimens. Tumor invasion of vascular structures adjacent to the pancreas can be predicted with preoperative CT and should alert the surgeon that vascular resection may be required. Histologic evidence of tumor cell infiltration of vessel walls was present in the majority of the resected specimens. PMID- 10481117 TI - Therapy for microcirculatory disorders in severe acute pancreatitis: effectiveness of platelet-activating factor receptor blockade vs. endothelin receptor blockade. AB - Many of the complications of severe acute pancreatitis are the result of the amplifying effects of microcirculatory disruption. The factors causing microcirculatory disorders in acute pancreatitis involve vasoactive mediators such as platelet-activating factor (PAF) and endothelin-1 (ET) activated during the inflammatory response to pancreatic injury. To further evaluate the potential therapeutic role of specific receptor antagonists (RA) to these mediators, the present study compares the effect of PAF and ET receptor blockade on microcirculation and organ function in a well-established rodent model of severe acute pancreatitis. Six hours after acute pancreatitis induction, rats were randomized to therapy with ET-RA (50 mg/kg LU-135252), PAF-RA (82 microg/kg WEB 2170), or NaCl 0.9% (volume equivalent). After 18 hours of fluid resuscitation, animals were relaparotomized for intravital microscopic determination of capillary blood flow, leukocyte rolling, and capillary permeability in the pancreas and colon. Other measurements included cardiorespiratory parameters, hematocrit, pleural effusions, ascites, urine production, and survival. Compared to saline treatment both ET-RA and PAF-RA significantly improved capillary blood flow in the pancreas and colon, reduced leukocyte rolling, and stabilized capillary permeability. The beneficial effects of receptor antagonist treatment on microcirculation were associated with decreased fluid loss into the third space, improved renal and respiratory function, and survival. Although both receptor antagonists likewise improved capillary blood flow, ET-RA was significantly more effective in counteracting leukocyte rolling and capillary leakage, thereby further reducing fluid sequestration. The present study confirms the beneficial effects of PAf and ET receptor blockade on microcirculation inside and outside the pancreas, organ function, and survival when given at the early stage of severe pancreatitis. Because ET-RA was more effective in stabilizing capillary permeability and avoiding subsequent fluid loss into the third space, we propose that ET-RA should be tested in a clinical trial (either in comparison or in addition to PAF-RA). PMID- 10481118 TI - Early increase in intestinal permeability in patients with severe acute pancreatitis: correlation with endotoxemia, organ failure, and mortality. AB - Sepsis accounts for 80% of deaths from acute pancreatitis. This study aimed to investigate early changes in intestinal permeability in patients with acute pancreatitis, and to correlate these changes with subsequent disease severity and endotoxemia. The renal excretion of enterally administered polyethylene glycol (PEG) 3350 and PEG 400 was measured within 72 hours of onset of acute pancreatitis to determine intestinal permeability. Severity was assessed on the basis of APACHE II scores and C-reactive protein measurements. Serum endotoxin and antiendotoxin antibodies were measured on admission. Eight-five patients with acute pancreatitis (mild in 56, severe in 29) and 25 healthy control subjects were studied. Urinary excretion of PEG 3350 (median) was significantly greater in patients who had severe attacks (0.61%) compared to those with mild disease (0.09%) and health control subjects (0.12%) (P <0. 0001), as was the permeability index (PEG 3350/400 excretion) (P <0. 00001). The permeability index was significantly greater in patients who subsequently developed multiple organ system failure and/or died compared with other severe cases (0.16 vs. 0.04) (P = 0.0005). The excretion of PEG 3350 correlated strongly with endotoxemia (r = 0.8; P = 0.002). Early increased intestinal permeability may play an important role in the pathophysiology of severe acute pancreatitis. Therapies that aim to restore intestinal barrier function may improve outcome. PMID- 10481120 TI - Force-feedback grasper helps restore sense of touch in minimally invasive surgery. AB - The age of minimally invasive surgery has brought forth astounding changes in the health care field. Less pain and quicker patient recovery have been demonstrated with several types of operations that were once performed by an open technique. With these changes have come reports of complications. The decreased sense of touch is just one of several limitations inherent to current techniques of minimally invasive surgery that limit detection of subtle or unapparent lesions on palpation, such as common duct stones and liver lesions. The purpose of this study is to demonstrate the ability of a force-feedback-equipped grasper to restore some of the sense of touch that is lost in minimally invasive surgery. To demonstrate this ability, we created six silicone phantoms of identical dimensions but graded compliance, and asked 10 subjects to place them in increasing/decreasing order of compliance. They used three tools (their dominant gloved hand, a standard laparoscopic Babcock grasper, and our force-feedback device fitted with the identical Babcock grasper) to rate the compliance of the samples in a blinded fashion. These conditions thus approximated the conditions of open surgery, minimally invasive surgery, and minimally invasive surgery fitted with a force-sensing device, in terms of palpating tissues. Five surgeons skilled in minimally invasive surgery and five nonsurgeons participated in the study. The results indicate that the force-feedback device is significantly (P <0.05) better than a standard Babcock grasper at rating tissue compliance, but was not as successful as a gloved hand (mean of squared errors = 1.06, 3. 15, and 0.25, respectively). There was no significant difference between surgeons and nonsurgeons in rating compliance. We conclude that this force-feedback instrument is able to partially restore the sense of touch in minimally invasive surgery. This restored ability may thus potentially result in more efficient operations with improved diagnostic capabilities and fewer complications during minimally invasive surgery. PMID- 10481119 TI - Prognostic factors in resectable pancreatic cancer: p53 and bcl-2. AB - The p53 tumor suppressor gene and the Bcl-2 proto-oncogene regulate cell cycle progression and apoptosis. We evaluated the expression of these molecular markers with standard pathologic prognostic variables in patients who received multimodality therapy for resectable adenocarcinoma of the pancreas to study the effect of p53 and Bcl-2 on survival duration. Immunohistochemical staining of archival material was performed to determine levels of expression of p53 and Bcl 2 proteins in 70 patients with adenocarcinoma of pancreatic origin. All patients underwent a potentially curative pancreaticoduodenectomy and standardized pathologic analysis of resected specimens. Potential pathologic and molecular prognostic variables were assessed for their effect on survival duration. Nuclear staining for p53 was observed in 33 (47%) of 70 specimens. Immunostaining for Bcl 2 was observed in 23 specimens (33%). A trend toward improved survival duration was seen in patients whose tumors stained positive for either p53 or Bcl-2. Negative staining for both markers predicted short survival (P = 0.01). By univariate and multivariate analyses, no single pathologic factor was associated with survival duration. Immunohistochemical staging using both p53 and Bcl-2 significantly predicted survival duration by univariate and multivariate analysis; patients whose tumors stained positively for p53 and/or overexpressed Bcl-2 had a significantly longer survival than those whose tumors stained negative for both proteins. PMID- 10481121 TI - Laparoscopic cholecystectomy in patients with hepatic cirrhosis: a five-year experience. AB - Our institution is a tertiary referral center that specializes in hepatobiliary surgery. To evaluate the safety, efficacy, and conversion rate of laparoscopic cholecystectomy in patients with hepatic cirrhosis, we conducted a retrospective analysis of all cirrhotic patients undergoing attempted laparoscopic cholecystectomy during the period from 1991 to 1996. The diagnosis of cirrhosis was made on the basis of either a preoperative history, a liver biopsy, or the surgeon's operative description of the liver. All patients had early, well compensated cirrhosis (Child's class A or B). A total of 30 patients underwent attempted laparoscopic cholecystectomy and five patients were converted to an open procedure (17%). The conversion rate for elective cases was 5% compared with 36% for urgent procedures. Two patients were converted because of varices and three because of unclear anatomy. No patients were converted because of bleeding. There were no operative deaths. The complication rate for elective procedures was 16%, with an average length of stay of 2.1 days, compared with 36% and 4.8 days, respectively, for urgent cases. Laparoscopic cholecystectomy in patients with early, well-compensated cirrhosis is safe and should be the treatment of choice for these patients. PMID- 10481122 TI - Multivariate analysis of factors predicting outcome after laparoscopic Nissen fundoplication. AB - Laparoscopic Nissen fundoplication has been applied with increasing frequency in the treatment of gastroesophageal reflux disease. The aim of this study was to determine the variables that predict outcome of laparoscopic Nissen fundoplication. A multivariate analysis was performed on data from 199 consecutive patients undergoing laparoscopic Nissen fundoplication. Variables included age, sex, body mass index, primary symptoms, clinical response to acid suppression therapy, erosive esophagitis, 24-hour esophageal pH score, and the percentage of time the esophageal pH was less than 4 on 24-hour pH monitoring, lower esophageal sphincter competence, status of the esophageal body motility, hiatal hernia, carditis, intestinal metaplasia of cardiac epithelium limited to the gastroesophageal junction, and Barrett's esophagus of any length. Clinical outcome was obtained from all patients at a median follow-up of 15 months (range 6 to 74 months) after surgery. One hundred seventy-three patients had an excellent or good outcome (87%) and 26 had a fair or poor outcome. Three factors were significantly predictive of a successful outcome: an abnormal 24-hour pH score (odds ratio = 5.4; 95% confidence interval [CI] = 1. 9-15.3), a typical primary symptom (odds ratio = 5.1; 95% CI = 1. 9-13.6), and a clinical response to acid suppression therapy (odds ratio = 3.3; 95% CI = 1.3-8.7). We conclude that 24-hour pH monitoring provides the strongest outcome predictor of laparoscopic Nissen fundoplication and that outcome is based more on the correct identification of the disease than on its severity. PMID- 10481123 TI - Gadolinium chloride inhibits lipopolysaccharide-induced mortality and in vivo prostaglandin E2 release By splenic macrophages. AB - The monocytic phagocytic system, consisting primarily of tissue macrophages of the liver and spleen, produces prostaglandin E(2) (PGE(2)), a modulator of the septic response. Macrophages are known to internalize gadolinium chloride (GD), a lanthanide metal, which inhibits phagocytic function. Thus we studied the effect of in vivo GD on lipopolysacchride (LPS)-induced mortality and on LPS-stimulated PGE(2) release by cultured splenic macrophages. GD (7 mg/kg intravenously) given on the two days prior to LPS challenge (30 mg/kg intravenously) completely prevented the uniform mortality in rats. This protective effect was transient since rechallenge with LPS 10 days later was uniformly lethal. Previous work in this laboratory has established a critical role of arginine concentration on macrophage behavior in vitro. Therefore, to establish culture conditions reflective of the milieu within the portal venous system, alanine and arginine levels were measured in the portal and hepatic veins of normal and endotoxemic (LPS, 10 mg/kg intraperitoneally) rats. In contrast to alanine levels, which were not altered by endotoxemia, there was a reduction of arginine concentrations from a range of 50 to 250 micromol/L in normal rats to a range of 10 to 50 micromol/L after LPS challenge. Consequently subsequent in vitro assays of splenic macrophage secretory behavior were performed in concentrations of 1200 micromol/L arginine (in standard RPMI-1640), as well as in concentrations reflective of physiologic arginine levels (10 and 100 micromol/L in modified RPMI-1640). Rat splenic macrophages harvested after two consecutive days of either in vivo saline or GD injection (7 mg/kg intravenously) were stimulated with LPS (0.025 to 2.5 microg/ml). At 72 hours of culture, the release of PGE(2) by splenic macrophages from GD-treated rats was significantly (P <0.0001) reduced at all LPS concentrations. Increased PGE(2) production was not present when the splenic macrophages were cultured in the supraphysiologic arginine (1200 micromol/L) concentration. The results demonstrate the relevance of physiologic arginine concentrations in cell culture studies and suggest that the protection conferred by GD against septic mortality may be related to downregulation of the release of immunosuppressive PGE(2) by the monocytic phagocytic system. PMID- 10481124 TI - Calcium accentuates injury induced by ethanol in human gastric cells. AB - The mechanism(s) whereby ethanol induces cellular injury remains poorly understood. Furthermore, the role of calcium in gastric mucosal injury under in vitro conditions is poorly defined. The major objectives of this study were to (1) define the temporal relationship between intracellular calcium accumulation induced by ethanol and cellular injury, (2) characterize the mechanism(s) whereby ethanol increases cellular calcium content, and (3) determine whether calcium removal would attenuate ethanol-induced cellular injury. Human gastric cells (AGS) were used for all experiments. Sustained intracellular calcium accumulation induced by ethanol, but not transient changes, preceded and directly correlated with cellular injury. Cells exposed to damaging concentrations of ethanol demonstrated an initial calcium surge that appeared to be a consequence of inositol 1,4,5-triphosphate (IP3) generation and subsequent internal store release followed by a sustained plateau resulting from extracellular calcium influx through store-operated calcium channels. Finally, both morphologic (cellular injury) and functional (clearance of bovine serum albumin) changes induced by ethanol were significantly attenuated when extracellular Ca(+&plus) influx was prevented, and further decreased when intracellular Ca(++) stores were depleted. These data indicate that calcium plays a significant role in cellular injury induced by ethanol. PMID- 10481127 TI - Introduction to threat assessment PMID- 10481125 TI - Extracellular matrix modulates enterocyte growth via downregulation of c-jun but is independent of p21 and p27 expression. AB - Regulation of the intestinal crypt-villus axis is multifactorial and involves growth factors and extracellular matrix composition. Laminin, a component of the enterocyte basement membrane, induces enterocyte differentiation and inhibits proliferation. To investigate the mechanism of this observation, we examined the expression of cell cycle modulators in enterocytes cultured on laminin. IEC-6 enterocytes were cultured on collagen I or laminin for 24 hours in media with serum followed by 48 hours of culture in serum-free media. Cells were then stimulated with epidermal growth factor, and RNA and protein were extracted before and up to 18 hours after stimulation. c-jun mRNA expression and p21 and p27 protein expression were analyzed. Expression of c-jun was inhibited in cells grown on laminin as compared to collagen I. Expression of p21 and p27 was no different between cells grown on laminin or collagen I. The mechanism of enterocyte growth inhibition mediated by laminin involves downregulation of c-jun expression. In contrast, p21 and p27 levels were unaffected by extracellular matrix indicating that the changes in expression of these cyclin-dependent kinase inhibitors do not contribute to the effect of laminin on enterocyte proliferation. PMID- 10481128 TI - Dangerousness and disability as predictors of psychiatric patients' legal status. AB - In a sample of patients admitted to two state psychiatric facilities, discriminant analyses were used to predict (1) legal status at admission (voluntary versus emergency detention), and (2) the subsequent decision to commit patients initially admitted under an order of emergency detention (court commitment versus release). Measures of preadmission dangerousness, followed by variables reflecting degree of disability or impairment, accounted for most of the variance in legal status at admission. Personal resources and demographic characteristics added little to the discrimination. Measures of disability accounted for most of the variance in the later decision to commit, whereas indices of dangerousness, personal resources, and demographics added little to discrimination of discharged and court-committed patients. These findings reflect the gap between legal standards and the practice of civil commitment, and support the argument that degree of disability plays a more important role than dangerousness in decisions to extend the hospitalization of involuntary patients. PMID- 10481126 TI - Cumulative risk of developing polyps or malignancy at the ileal pouch-anal anastomosis in patients with familial adenomatous polyposis. AB - Restorative proctocolectomy with an ileal pouch-anal anastomosis is performed in an increasing number of patients with familial adenomatous polyposis (FAP). Two techniques are currently used to construct an ileal pouch-anal anastomosis: (1) a double-stapled anastomosis between the pouch and the anal canal and (2) mucosectomy with a hand-sewn ileoanal anastomosis at the dentate line. Although this procedure is thought to abolish the risk of colorectal adenoma, an increasing number of case reports have been published concerning the development of adenoma at the anastomotic site. The purpose of this study was to evaluate the overall cumulative risk of developing adenomatous polyps after ileal pouch-anal anastomosis and to compare the cumulative risk after either anastomotic technique. A total of 126 consecutive FAP patients undergoing a restorative proctocolectomy were identified from polyposis registries in The Netherlands, Denmark, Italy, Germany, and New York. Life-table analysis was used to calculate the cumulative risk of developing polyps in 97 patients with at least 1 year of endoscopic follow-up (median 66 months, range 12 to 188 months). A double-stapled anastomosis was used in 35 patients, whereas in 62 patients a hand-sewn anastomosis with a mucosectomy was performed. In 13 patients polyps developed at the anastomotic site, four with severe and four with moderate dysplasia. None of the patients developed a carcinoma at the anastomotic site. The cumulative risk of developing a polyp at the anastomotic site was 8% (95% confidence interval 2% to 14%) at 3.5 years and 18% (95% confidence interval 8% to 28%) at 7 years, respectively. The risk of developing a polyp at the anastomotic site within 7 years was 31% for patients with a double-stapled vs. 10% for patients with a hand sewn anastomosis with mucosectomy (P = 0.03 [log-rank test]). Because FAP patients undergoing a restorative proctocolectomy with either a double-stapled or hand-sewn anastomosis have a substantial risk of developing adenomatous polyps at the anastomotic site, lifelong endoscopic surveillance is mandatory in both groups. PMID- 10481129 TI - The dangerous nature of intimate relationship stalking: threats, violence, and associated risk factors. AB - Although previous studies of forensic and law enforcement stalking populations (Harmon, Rosner, & Owens, 1995; Meloy & Gothard, 1995; Zona, Sharma, & Lane, 1993) agree that the degree of intimacy of the victim-suspect relationship is an important factor in stalking cases, they have not conducted in-depth analyses of this variable. This study compared 223 intimate (n=135) and non-intimate (n=88) stalking cases managed by the Los Angeles Police Department's Threat Management Unit. A path analysis revealed a significant relationship between the stalkers' intimate versus non-intimate status and violence committed toward persons and property. This relationship was positively influenced by the suspect's level of proximity to the victim and threats toward the victim and property, but not influenced by suspect's criminal, psychiatric, and domestic violence histories. Overall, intimate relationship stalkers used more dangerous stalking behaviors than non-intimate relationship stalkers. Risk factors for assessing dangerousness of stalkers are discussed. PMID- 10481130 TI - Aggressive acts and assaults in intimate relationships: towards an understanding of the literature. AB - Far more people in relationships are subjected to violent acts than those who receive injuries. The degree of damage sustained may not reflect the perpetrator's intent to deliberately harm a partner. Data documenting aggressive acts determines the population at risk and their prevention and early treatment requirements; whereas data focusing on harm and injury helps determine emergency medical and refuge services. Data from national crime surveys, police records, or clinical populations should not be generalized to the population at large. Even if men perpetrate the majority of serious partner attacks, addressing the issue of female violence will significantly reduce the overall level of domestic violence. Judicial, medical, and social services should take note that while male violence may be more problematic, violence is a relationship issue, not a male issue. PMID- 10481131 TI - Developing profiles of risk for parental abduction of children from a comparison of families victimized by abduction with families litigating custody. AB - This study systematically compared parents in abducting families with families litigating custody. Findings indicate that risks for parental abduction of children are multidetermined by: (1) a heightened concern about very young children being exposed to neglectful, endangering, or criminal environments by the other parent; (2) unsubstantiated allegations of sexual abuse; (3) heightened distrust of and less respect for law and authority; and (4) a reluctance to seek help from the courts. Abducting families were also predominantly socially and economically disadvantaged: parents were less likely to have been married to one another; they had lower incomes, were more poorly educated, and were disproportionately members of minority racial and ethnic groups. The social policy dilemmas of identifying these differences as risk factors are discussed together with suggestions for risk management. PMID- 10481132 TI - Threat assessment: defining an approach for evaluating risk of targeted violence. AB - Although the field of risk assessment has made tremendous advances in the past 20 years, assessments of targeted violence continue to pose a significant challenge to law enforcement, mental health, and other professionals. These specific and critical assessments require an innovative approach. The threat assessment model, developed and refined by the U.S. Secret Service, provides a useful framework for thinking about assessments of potential for targeted violence. In this paper, we attempt to define this approach as it has been developed by the Secret Service, and apply it within the existing professional/scientific literature on risk assessment. We begin with a brief review of existing models and approaches in risk assessment, and identification of some gaps in our existing knowledge as it relates to assessments of targeted violence. We then proceed with an outline of the threat assessment approach, including a review of principles and guiding operational questions, and discussion of its use in assessment of targeted violence. PMID- 10481133 TI - Assessing threats of targeted group violence: contributions from social psychology. AB - Recent increases in domestic and international acts of extremist violence perpetrated against American citizens have prompted an increased need for information to help understand and evaluate the threat posed to U.S. targets by extremist groups and their individual members. The purpose of this paper is to (i) suggest the potential relevance of social psychological research on group behavior for understanding and assessing threats of extremist group violence; and, (ii) encourage more systematic research on group violence to further inform assessments of group risk. Approaching the issue from the levels of group behavior, and of individual behavior within a group context, the article summarizes research on key principles of group behavior, and the effects of group membership on individual behavior; proposes specific questions derived from these principles for consideration in evaluating risk for violence by groups, and by individuals influenced by groups; and suggests further research needs. PMID- 10481134 TI - Suicide and violence risk in law enforcement: practical guidelines for risk assessment, prevention, and intervention. AB - Research and anecdotal reports indicate that suicide and violence risk may be higher among members of law enforcement than those in other occupational categories. This article examines the phenomenon of suicide and violence risk within this population, and law enforcement cultural variables that may contribute to elevated risk. Suicide and violence risk factors and clues unique to law enforcement are described, as are intervention approaches which may be helpful to managing and reducing risk. PMID- 10481135 TI - Empirically identifying factors related to violence risk in corrections. AB - The authors used structural modeling to predict institutional aggression among male mentally ill offenders using the predictors of anger, antisocial personality style, current violent offense, ethnicity, and impulsivity. Measures included the Barratt Impulsiveness Scale, the Buss-Perry Aggression Questionnaire, the Personality Assessment Inventory, age, ethnicity, current violent offense, victim injury from current offense, and institutional incidents of physical and verbal aggression. The model fit the data, and accounted for 94% and 87% of the variance of physical and verbal aggression, respectively. Results indicated anger, antisocial personality style, and impulsivity are stronger predictors of institutional aggression than are ethnicity and current violent offense; anger was the best predictor. Results suggest dynamic variables such as anger can be targeted for clinical intervention to reduce institutional violence. PMID- 10481137 TI - John C. "Chris" hatcher (1946-1999) PMID- 10481136 TI - Differentiating stranger murders: profiling offender characteristics from behavioral styles. AB - The literature on "offender profiling" suggests that different "styles" of homicide will reflect differences in the background characteristics of the offender. To test this suggestion, hypotheses were drawn from studies of aggression to propose that murder crime scenes would reveal stylistic distinctions in the role of the aggression in the offense. It was further hypothesized that these distinctions would be clearest for those crime scene indicators that reflect the instrument (cognitive) actions that shape the offense rather than the more expressive (impulsive and emotional) components. Hypotheses about associated offender characteristics were also deduced on the assumption that the murder scene theme revealed the killers' typical styles of interpersonal transaction. To test these hypotheses MDS analyses were carried out on the crime scene and offender characteristics derived from 82 single offender, single victim stranger homicides. A multivariate structure resulted, including all three hypothesized styles, allowing 65% of the cases to be assigned to unique styles and a further 36% to be assigned to appropriate hybrids. Offender characteristics related in the anticipated way to the different crime scene styles, providing a basis for law enforcement inferences about offenders in stranger murders. PMID- 10481139 TI - Rediscovering natural product biodiversity. PMID- 10481138 TI - How many leads from HTS? PMID- 10481140 TI - Expediting the preclinical development process. PMID- 10481141 TI - The cell cycle and drug discovery: the promise and the hope. AB - In recent years, there have been major developments in the understanding of the cell cycle. It is now known that normal cellular proliferation is tightly regulated by the activation and deactivation of a series of proteins that constitute the cell cycle machinery. The expression and activity of components of the cell cycle can be altered during the development of a variety of diseases where aberrant proliferation contributes to the pathology of the illness. Apart from yielding a new source of untapped therapeutic targets, it is likely that manipulating the activity of such proteins in diseased states will provide an important route for treating proliferative disorders, and the opportunity to develop a novel class of future medicines. PMID- 10481142 TI - Use and value of metabolism databases. AB - A key objective during drug discovery is to ensure selection of lead compounds for development that have the optimum pharmacodynamic profile without undesirable toxicity. Metabolism plays a key role in determining the biological activity of compounds in vivo, and is therefore an important factor to consider during the discovery phase. The ability to predict the biotransformation pathways for specific chemical structures provides the opportunity to implement structural modifications that might advantageously modify these processes. Potentially valuable tools include knowledge databases that contain searchable information on the known metabolites of existing compounds and databases that are designed to predict the metabolites. PMID- 10481143 TI - Gene-regulating protein kinases as important anti-inflammatory targets. AB - Protein phosphorylation is a key cellular regulatory mechanism. Protein kinases and phosphatases regulate cell-cycle progression, transcription, translation, protein sorting and cell adhesion events that are critical to the inflammatory process. Two of the best- characterized immunosuppressants, cyclosporin and rapamycin, are also effective anti-inflammatory drugs. They act directly on protein phosphorylation and, as such, validate the concept that small-molecule modulators of phosphorylation cascades possess anti- inflammatory properties. The authors describe studies that outline progress in defining specific protein kinase signal-transduction cascades, the key drug discovery targets in these cascades and progress towards developing selective agents that have potential in treating numerous inflammatory diseases. PMID- 10481144 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 10481145 TI - Combinatorial chemistry. PMID- 10481146 TI - Bioinformatics. PMID- 10481147 TI - Renewed strategies for drug development against parasitic diseases. PMID- 10481148 TI - Livestock management and malaria prevention in irrigation schemes. PMID- 10481149 TI - The case for biological control of snail intermediate hosts of Schistosoma mansoni. PMID- 10481150 TI - Selection, distinction and semantics on the Web. PMID- 10481151 TI - Tsetse-trypanosome interactions: rites of passage. AB - Trypanosomes that cause sleeping sickness (Trypanosoma brucei rhodesiense and T. b. gambiense) are entirely dependent on tsetse for their transmission between hosts, but the flies are not easily infected. This situation has not arisen by chance - the tsetse has evolved an efficient defence system against trypanosome invasion. In this review, Susan Welburn and Ian Maudlin chart the progress of trypanosomes through the fly and identify some of the hazards faced by both parasite and fly that affect vector competence of tsetse. PMID- 10481152 TI - Enzymes of parasite thiol metabolism as drug targets. AB - The potential for chemotherapeutic exploitation of thiol metabolism in parasitic protozoa is reviewed here by Luise Krauth-Siegel and Graham Coombs. The review is based largely on discussions held at a meeting of the COST B9 Action entitled 'Chemotherapy of Protozoal Infections'*. The major questions posed were: which enzymes are the best to target; what further information is required to allow their use for rational drug development; and how can this be achieved most efficiently? Not surprisingly, only partial answers could be obtained in many cases, but the interactive discussion between the multidisciplinary group of participants provided thought-provoking ideas and will help direct future research. PMID- 10481153 TI - Infochemicals in mosquito host selection: human skin microflora and Plasmodium parasites. AB - The interaction between the African malaria vector Anopheles gambiae and its human host has traditionally been viewed within a bitrophic context, considering only the human and the mosquito. Recently, the influence of the Plasmodium parasite on the interaction has been recognized, because it affects the physiology and/or behaviour of humans and mosquitoes. However, studies on odour mediated host-seeking behaviour of An. gambiae and other Diptera have provided evidence that a fourth group of organisms should be taken into consideration. Human skin microflora play a role in the production of odorous compounds that might function as kairomones for mosquitoes. Here, Marieta Braks, Rob Anderson and Bart Knols introduce the role of human microflora into the process of odour mediated host selection and review the interaction in a multipartite context so as to identify research avenues that will enhance our limited knowledge of this aspect of malaria transmission. PMID- 10481154 TI - Websites of interest PMID- 10481155 TI - Parasitic nematodes, proteinases and transgenic plants. AB - Parasite proteinases have important functions in host-parasite interactions. Consequently, they have been investigated as targets for the control of both plant and animal parasites. Plant parasitic nematodes cause estimated annual losses to world agriculture of US$100 billion and, currently, their control often relies on highly toxic nematicides, with associated environmental risks. The potential of disrupting digestive proteinases for plant parasitic nematode control, via expression of proteinase inhibitors in transgenic plants, is summarized here by Catherine Lilley, Pauline Devlin, Peter Urwin and Howard Atkinson. They then consider whether the approach of expressing antinematode proteins in plants can be adapted for control of certain animal parasitic nematodes. PMID- 10481157 TI - How prevalent are Plasmodium ovale and P. malariae in East Asia? AB - Plasmodium ovale and Plasmodium malariae, two of the four human malaria parasites, are usually found at very low prevalence in East Asia, even in areas with intense malaria transmission. In this article, Fumihiko Kawamoto, Qing Liu, Marcelo Ferreira and Indah Tantular review data obtained in recent field surveys, using alternative diagnostic methods such as acridine orange staining and PCR based methods, to evaluate the prevalence of these two malaria species in East Asia. They argue that these species might be much more prevalent in East Asia than reported previously. In addition, they discuss the implications of sequence variations found in the small subunit ribosomal RNA genes of the two species targeted by diagnostic PCR and compare morphological criteria for speciation of malaria parasites stained with Giemsa and acridine orange. PMID- 10481156 TI - Cytokines, nitric oxide, heat shock proteins and virulence in Toxoplasma. AB - Elucidating the factors that play important roles in the expression of virulence by parasites is crucial to understanding disease pathogenesis and to developing control strategies rationally. Here, Kate Miller, Nick Smith and Alan Johnson, using Toxoplasma gondii as a model, argue that the interactions between the immune system and 70 kDa heat shock proteins of apicomplexan parasites profoundly influence parasite virulence. PMID- 10481158 TI - Some rethinking on bednets. PMID- 10481161 TI - Targeted Antiestrogens to Prevent Breast Cancer. AB - Tamoxifen, the first selective estrogen receptor modulator (SERM), has provided invaluable laboratory and clinical evidence that such drugs can reduce the risk of breast cancer and increase bone density. Raloxifene is available for the prevention of osteoporosis, and preliminary studies show a decrease in breast cancer risk. On the basis of current information about the pharmacology of SERMs, a large breast cancer prevention trial called study of tamoxifen and raloxifene (STAR) has started in the USA. Results should be available in the next seven years. PMID- 10481160 TI - The Molecular Pharmacology of SERMs. AB - Estrogen-containing medicines have been used successfully for the past 50 years for the treatment of conditions associated with menopause. Although initially considered a reproductive hormone, millions of years of clinical exposure to estrogen(s) have indicated that its influence extends to a variety of target tissues not generally considered to be involved in reproduction. Specifically, estrogen has positive actions in the skeleton, the cardiovascular system and possibly the central nervous system, activities that combine to have a positive impact on mortality and morbidity. However, despite the medical benefits afforded by estrogen replacement therapy, the number of women who initiate or remain on therapy for longer than one year is relatively small. This is due in part to the fear that estrogens increase the risk for breast cancer. Consequently, it was realized several years ago that novel estrogen receptor modulators were needed, which would retain the beneficial effects of estrogens in most target organs but be inactive in the breast. Although the perfect tissue-selective estrogen remains to be identified, progress in this direction has been made. In the past year, for example, we have seen selective estrogen receptor modulators (SERMs) enter into the clinic for the prevention of osteoporosis. Compounds of this class, which function as estrogens in the skeletal system but oppose estrogen action in the breast, are the first step in developing the perfect hormone replacement therapy. This review summarizes the complex pharmacology of the SERMs and illustrates how they differ mechanistically from estradiol, the physiological ligand of the estrogen receptor. PMID- 10481159 TI - Evolving concepts of selective estrogen receptor action: from basic science to clinical applications. PMID- 10481162 TI - SERM Drugs for the Prevention of Osteoporosis. AB - Although estrogens have proved useful in the prevention and treatment of osteoporosis, their side effects (for example, those on breast and endometrial cancer) are worrying to patients and physicians alike. Therefore, selective estrogen receptor modulator (SERM) drugs have been developed for use in their stead. The triphenylethylene drug tamoxifen proved to be protective against bone loss, but had side effects on uterus similar to those of natural estrogens. The tamoxifen derivative toremifene has less effect on bone. Further derivatives of tamoxifen (droloxifene, idoxifene) can be expected to act like tamoxifen when approved for clinical testing. The non-steroidal benzothiophene derivative, raloxifene, is the best SERM available thus far. It does not increase the incidence of endo-metrial cancer; in addition, like tamoxifen, it has the potential to prevent breast cancer, but has a better profile in its actions on bone (for example, it reduces the vertebral fracture rate more effectively than tamoxifen). Unlike estrogen, it decreases blood triglicerides as well as cholesterol. PMID- 10481163 TI - The Potential of SERMs for Reducing the Risk of Coronary Heart Disease. AB - Selective estrogen receptor modulators (SERMs) represent a growing class of compounds that act as either estrogen receptor agonists or antagonists in a tissue-selective manner. Preclinical and clinical studies have shown that estrogen has favorable effects on serum lipids and might affect processes at the blood vessel wall to inhibit atherosclerosis. SERMs with the appropriate selectivity profile offer the opportunity to dissociate these favorable cardiovascular effects of estrogen from its unfavorable stimulatory effects on the breast and uterus. This article reviews the data from both animal and human studies that document the cardiovascular effects of SERMs and discusses the clinical implications of these results. PMID- 10481164 TI - Transcription Factors and Hypopituitarism. AB - Several homeodomain factors are found in the developing anterior pituitary lobe. The production of these developmental transcription factors has distinct temporal and spatial patterns. By interacting with each other, as well as with other extrinsic and intrinsic signals, they control cell determination and specification. Here, we discuss transcription factors that have been shown to have an in vivo role in pituitary cell-type specification. PMID- 10481167 TI - Glucocorticoid Receptor beta: View II. AB - There is a clear role for mechanisms that modulate glucocorticoid receptor (GR) function. The non-steroid-binding GRbeta isoform has been proposed to play a role in this modulation but the published data are contradictory. The relative levels of this isoform appear to be low. Alternative mechanisms for the modulation of glucocorticoid action are described and contrasted with the proposed role for GRbeta. PMID- 10481166 TI - Glucocorticoid Receptor beta: View I. AB - Since the human glucocorticoid receptor (GR) cDNA and gene sequences were reported, the existence of two highly homologous GR isoforms was predicted. These were the classic human ligand-binding GRalpha, and a slightly smaller protein, termed GRbeta. Although the mechanism of action of GRbeta has been studied extensively, the role of GRbeta in the modulation of glucocorticoid actions remains uncertain. PMID- 10481168 TI - The cell factory needs a model of a factory. PMID- 10481169 TI - Laser-guided direct writing for applications in biotechnology. AB - Laser-induced optical forces can be used to guide and deposit 100 nm - 10 microm diameter particles onto solid surfaces in a process we call 'laser-guided direct writing'. Nearly any particulate material, including both biological and electronic materials, can be manipulated and deposited on surfaces with micrometer accuracy. Potential applications include three-dimensional cell patterning for tissue engineering, hybrid biological-electronic-device construction, and biochip-array fabrication. PMID- 10481170 TI - Colloidal gas aphrons: potential applications in biotechnology AB - Colloidal gas aphrons are microbubbles encapsulated by surfactant multilayers. They provide a large interfacial area to adsorb charged and/or hydrophobic molecules; the extent and mechanism of the adsorption depends on the surfactant multilayer. The physical properties of colloidal gas aphrons have recently been characterized for a range of surfactants in order to find the best systems for particular applications. A range of exciting biotechnology applications has been identified, including the recovery of cells, proteins and other biological molecules, and the enhancement of gas transfer in bioreactors and bioremediation. PMID- 10481171 TI - Application of in situ product-removal techniques to biocatalytic processes. AB - Biocatalytic processes for the manufacture of small, highly functionalized molecules frequently have limited productivity. A common reason for this is the presence of the reaction products that can cause inhibitory or toxic effects (making poor use of the enzyme) or promote unfavourable equilibria (giving low conversions). In each case, the product needs to be removed as soon as it is formed in order to overcome these constraints and hence increase the productivity of the biocatalytic process. Here, we review the need for in situ product removal and the process research required for its implementation. PMID- 10481172 TI - The Earth's bounty: assessing and accessing soil microbial diversity. AB - The study of microbial diversity represents a major opportunity for advances in biology and biotechnology. Recent progress in molecular microbial ecology shows that the extent of microbial diversity in nature is far greater than previously thought. Here, we discuss methods to analyse microorganisms from natural environments without culturing them and new approaches for gaining access to the genetic and chemical resources of these microorganisms. PMID- 10481173 TI - Rationalizing the design of polymeric biomaterials. AB - Polymers are a promising class of biomaterials that can be engineered to meet specific end-use requirements. They can be selected according to key 'device' characteristics such as mechanical resistance, degradability, permeability, solubility and transparency, but the currently available polymers need to be improved by altering their surface and bulk properties. The design of macromolecules must therefore be carefully tailored in order to provide the combination of chemical, interfacial, mechanical and biological functions necessary for the manufacture of new and improved biomaterials. PMID- 10481174 TI - Titin: a molecular control freak. AB - Recent studies of the giant protein titin have shed light on its roles in muscle assembly and elasticity and include the surprising findings described here. We now know that the titin kinase domain, which has long been a puzzle, has a novel regulation mechanism. A substrate, telethonin, has been identified that is located over one micron away from the kinase domain in mature muscle. Single molecule studies have demonstrated the fascinating process of reversible mechanical unfolding of titin. Lastly, and most surprisingly, it has been claimed that titin controls assembly and elasticity in chromosomes. PMID- 10481175 TI - Shuffling the deck: plant signalling plays a club. AB - The transmission of signals across the plasma membrane of cells plays an integral part in cell communication in unicellular and complex organisms. Protein kinases and their activators serve key roles in this process, and a number of paradigms have been established to describe their mode of action. Signalling in plant cells appears to shuffle these paradigms - as evidenced by two recent reports on the development of the Arabidopsis meristem. PMID- 10481176 TI - Septins: cytoskeletal polymers or signalling GTPases? AB - Septins are a family of conserved proteins that have been implicated in a variety of cellular functions involving specialized regions of the cell cortex and changes in cell shape. The biochemistry and localization of septins suggest that they form a novel cytoskeletal system or that they function as scaffolds for the assembly of signalling complexes. This article discusses septin biochemistry and septin-interacting proteins, focusing on the missing link between the structure and biochemical properties of septin proteins, and on how they function at a molecular level in processes such as cytokinesis and yeast budding. PMID- 10481177 TI - Protein translocation in apoptosis. AB - In programmed cell death (apoptosis), receptor-generated or other signals are transmitted to all cellular compartments, resulting in an apoptotic cell with extensive cytoplasmic and nuclear alterations. Protein translocation is now recognized as being crucial in the induction, amplification and regulation of this process. Diverse mechanisms trigger protein translocation to and from the plasma membrane, mitochondrion and nucleus during apoptosis. This review discusses where, why and how the various protein-translocation events take place and highlights their importance in the execution and regulation of apoptosis. PMID- 10481178 TI - Bacterial type II protein export and pilus biogenesis: more than just homologies? AB - Protein export by Gram-negative bacteria requires devoted machineries to allow for the passage of hydrolytic enzymes and toxins through the cell envelope. The Type II export machinery has a number of distinct characteristics, which include its role as an extension of Sec-dependent secretion, its ability to recognize and export fully folded substrates efficiently and, perhaps most significantly, the relationship between a subset of its gene products with the Type IV pilus biogenesis apparatus. An important question is whether we can extrapolate our knowledge, albeit limited, of Type IV pilus biogenesis to understand the structure and function of the Type II export apparatus. This and other questions relating to the energetics of assembly and specificity of the apparatus are addressed in this article. PMID- 10481179 TI - Unravelling the kinetoplastid paraflagellar rod. AB - Researchers who study human pathogens are often interested in unique and essential aspects of the biology of the pathogen. Recent progress has been made in understanding such a target in kinetoplastid parasites. The paraflagellar rod is a unique cytoskeletal structure that plays a key role in the life-cycle of these fascinating organisms. This review discusses the protein components and structure of the paraflagellar rod and its function in cell motility. PMID- 10481180 TI - George E. Palade: charting the secretory pathway. PMID- 10481181 TI - Careers-perspective interview PMID- 10481182 TI - Neuronal apoptosis in acute necrotic insults: why is this subject such a mess? AB - It is now recognized that necrotic neurological insults often trigger apoptosis in a subset of neurons. It is also now apparent that such apoptosis rarely matches the 'classical' apoptosis seen during development or the physiological turnover of cells outside the nervous system. As a result, the view has emerged that the 'apoptosis-like' changes that follow necrotic insults represent a different phenomenon, which is on a vague continuum with the necrotic features of cell death. We suggest that apoptosis following neurological insults is, in actuality, mechanistically identical to classical apoptosis. However, the atypical apoptotic endpoints that are observed are inevitable, given the way in which insult-triggered apoptosis is likely to have evolved. PMID- 10481183 TI - Stress and cognition: are corticosteroids good or bad guys? AB - Corticosteroid hormones secreted by the adrenal cortex protect the brain against adverse events and are essential for cognitive performance. However, in recent literature, the central action of corticosteroids has mostly been portrayed as damaging and disruptive to memory formation. We argue that this paradox can be explained by appreciating the specific role of both mineralocorticoid and glucocorticoid receptors in the various stages of information processing. In addition, the context in which corticosteroid-receptor activation takes place is crucial in determining steroid-mediated effects. These effects generally favour adaptive behaviour that is most relevant to the situation. Corticosteroid effects on cognition can, however, turn from adaptive into maladaptive, when actions via the two corticosteroid-receptor types are imbalanced for a prolonged period of time. PMID- 10481184 TI - FENS 2000 - Federation of European Neuroscience Societies millennium meeting. AB - In two articles by Wolf Singer, published in TINS in 1994 and 1997, brief analyses were presented of the problems faced by the European neuroscience community in its attempt to provide a cohesive response to meet the challenge of the proclamation of the 'Decade of the Brain'. One of the most-important decisions was to promote a forum for European neuroscience that, among other activities, would hold bi-annual meetings, alternating with meetings of partner European neuroscience societies in the intervening year. The first fruit of this was the enormously successful meeting held in Berlin in June 1998. The next meeting, the FENS millennium meeting, will be held in Brighton, UK (24-28 June 2000). PMID- 10481185 TI - 'Psychosurgery' in renaissance art. AB - Hieronymus Bosch and other early Renaissance artists depicted 'stone operations' in which stones were supposedly surgically removed from the head as a treatment for mental illness. These works have usually been interpreted either as portraying a contemporary practice of medical charlatans or as an allegory of human folly, rather than a real event. As trepanation for head injury and mental disease was actually carried out in Europe at this time, another interpretation of these works is that they are derived from a common medical practice of the day. PMID- 10481186 TI - Comparative studies of sex differences in the song-control system of songbirds. AB - Songbirds exhibit some of the most extreme sex differences in the brain of all vertebrates. Understanding the function of these sex differences has relied on making interspecies comparisons. In some species, females sing rarely or not at all, and the brain nuclei that control song are many times larger in volume in males than in females. In other species, males and females sing approximately equally, and the sizes of the brain nuclei that control song are approximately equal between the sexes. This article reviews sex differences in the song-control system of songbirds, and introduces statistical comparative methods developed by evolutionary biologists. These methods control for phylogenetic effects while comparing the co-evolution of traits. The extreme sex differences in song seem to have co-evolved with the extreme sex differences in singing behavior in songbird species. PMID- 10481187 TI - Time to decrease the weight attached to looming? PMID- 10481189 TI - Reply. PMID- 10481188 TI - The many ways of building collision-sensitive neurons. PMID- 10481190 TI - Presenilins: molecular switches between proteolysis and signal transduction. AB - Mis-sense mutations of presenilin 1 increase the release of amyloidogenic peptide from amyloid precursor protein (APP) and are a major cause of familial Alzheimer's Disease. Loss-of-function mutations of presenilins in the mouse, Caenorhabditis elegans and Drosophila result in severe developmental defects caused by disturbed Notch signalling. Recent studies suggest that the diverse biological roles of presenilin 1 can be explained at the molecular level by its role in the proteolytic cleavage of the integral membrane domains of Notch and APP. This cleavage is a central switch in Notch signalling, while, for APP, its physiological role remains elusive. Evidence that presenilin 1 itself has catalytic properties could explain many of the biological and biochemical alterations caused by presenilin-1 deficiency or clinical mutations in presenilin 1. However, as presenilins reside in the endoplasmic reticulum and the cleavage of Notch and APP is believed to occur close to the cell membrane, the scientific field now faces a 'spatial paradox'. PMID- 10481191 TI - Binding of signals relevant for action: towards a hypothesis of the functional role of the pontine nuclei. AB - If numbers matter, the projection that connects the cerebral cortex to the cerebellum is probably one of the most-important pathways through the CNS. Its extensive development as one ascends the phylogenetic scale parallels that of the cerebral hemispheres and the cerebellum, and it accompanies improvements in motor skills, suggesting that this system might have a decisive role in the generation of skilled movement. This article focuses on the pontine nuclei (PN), which are intercalated in the cerebro-cerebellar pathway, a large nuclear complex in the ventral brainstem of mammals, whose raison d'etre has as yet not been examined. By considering recent morphological and electrophysiological findings, this article argues that the PN are an interface that is needed to accommodate the grossly different computational principles governing the cerebral cortex and the cerebellum. PMID- 10481192 TI - Shaping excitation at glutamatergic synapses. AB - Glutamatergic synapses vary, exhibiting EPSCs of widely different magnitudes and timecourses. The main contributors to this variability are: presynaptic factors, including release probability, quantal content and vesicle composition; factors that modulate the concentration and longevity of glutamate in the cleft, including diffusion and the actions of glutamate transporters; and postsynaptic factors, including the types and locations of ionotropic glutamate receptors, their numbers, and the nature and locations of associated intracellular signalling systems. PMID- 10481193 TI - Protein phosphorylation and the regulation of synaptic membrane traffic. AB - It is well established that protein phosphorylation has an important role in synaptic plasticity. This is achieved, in part, via the presynaptic modulation of neurotransmitter release by protein kinases and protein phosphatases. In recent years, the increase in information available about proteins that are involved in synaptic exocytosis and endocytosis has been exploited in order to study the effects of protein phosphorylation on synaptic-vesicle cycling at the molecular level. The best-characterized protein in this respect is synapsin, whose function in the release of synaptic vesicles from the reserve pool is regulated by phosphorylation. More recently, it has emerged that proteins that function at other stages of the synaptic-vesicle cycle, which include priming of vesicles for docking-fusion and endocytic recycling, are also controlled by phosphorylation. Furthermore, recent work suggests that this regulation of membrane traffic by phosphorylation also occurs postsynaptically, where it contributes to synaptic plasticity. PMID- 10481194 TI - Parallel neural networks for learning sequential procedures. AB - Recent studies have shown that multiple brain areas contribute to different stages and aspects of procedural learning. On the basis of a series of studies using a sequence-learning task with trial-and-error, we propose a hypothetical scheme in which a sequential procedure is acquired independently by two cortical systems, one using spatial coordinates and the other using motor coordinates. They are active preferentially in the early and late stages of learning, respectively. Both of the two systems are supported by loop circuits formed with the basal ganglia and the cerebellum, the former for reward-based evaluation and the latter for processing of timing. The proposed neural architecture would operate in a flexible manner to acquire and execute multiple sequential procedures. PMID- 10481196 TI - How birds became airborne. PMID- 10481195 TI - AIDS and the brain: is there a chemokine connection? AB - Many HIV-1-positive individuals suffer from a variety of neurological problems known collectively as the HIV-1-related cognitive-motor complex. However, the molecular mechanisms that underlie HIV-1-induced neuropathology are unclear. They might include a combination of indirect effects, which result from the release of neurotoxins from activated astrocytes and microglia, and the direct effects of HIV-1-related proteins, such as gp120, on neurons. As the interaction of gp120 with immune cells has been shown to require the participation of chemokine receptors, this article explores the possibility that such receptors participate in the events underlying HIV-1-induced neuropathology. It is now clear that many types of cell in the brain possess chemokine receptors, including microglia, glia and neurons, and the interaction of gp120 with neuronal chemokine receptors initiates apoptotic death of neurons in vitro. Such effects might be modified by the actions of chemokines that act at these same receptors. However, the importance of this direct interaction with neurons in vivo and its relevance in the pathogenesis of AIDS-related dementia still needs to be established. Furthermore, the existence of chemokine receptors on neurons suggests that chemokines might regulate neuronal functions physiologically. PMID- 10481198 TI - What do impact factors tell us? PMID- 10481197 TI - Mosaic landscapes, family structure and evolution. PMID- 10481199 TI - Vascular plant controls on methane emissions from northern peatforming wetlands. AB - Methane emissions from wetlands are highly variable, both spatially and temporally and at scales ranging from microtopographic to regional differences. To comprehend this variation fully and also to predict responses to climate change, an understanding of the intimate linkage between carbon cycling and methane emission in these systems is needed. The presence of vascular plants has been recognized recently as one of the key factors controlling the scale of methane fluxes because it affects processes coupled to transport, production and consumption of methane. A wide area of research has therefore opened up, calling for investigations into details of the impact of vascular plants on methane emissions. PMID- 10481200 TI - AFLP genotyping and fingerprinting. AB - Amplified fragment length polymorphisms (AFLPs) are polymerase chain reaction (PCR)-based markers for the rapid screening of genetic diversity. AFLP methods rapidly generate hundreds of highly replicable markers from DNA of any organism; thus, they allow high-resolution genotyping of fingerprinting quality. The time and cost efficiency, replicability and resolution of AFLPs are superior or equal to those of other markers [allozymes, random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP), microsatellites], except that AFLP methods primarily generate dominant rather than co-dominant markers. Because of their high replicability and ease of use, AFLP markers have emerged as a major new type of genetic marker with broad application in systematics, pathotyping, population genetics, DNA fingerprinting and quantitative trait loci (QTL) mapping. PMID- 10481201 TI - Mitogenomics: digging deeper with complete mitochondrial genomes. AB - Mitochondrial genomes are being used to study increasingly ancient divergences among animal groups. Recent studies of complete mitochondrial DNA sequences have arrived at somewhat heretical conclusions, raising questions about the use of mitochondrial gene sequences for studying the relationships among highly divergent lineages. Other studies have documented convergent evolution of mitochondrial gene order, casting doubt on the use of these characters for phylogenetic analysis. The use of mitochondrial genomes for studying such deep divergences is coming under increased scrutiny, and these novel results need to be confirmed with data from nuclear genes. PMID- 10481202 TI - Sexual selection: separating genes from imprinting. PMID- 10481203 TI - Synchronicity in population systems: cause and consequence mixed. PMID- 10481204 TI - Consequences of the Allee effect for behaviour, ecology and conservation. AB - Warder C. Allee brought attention to the possibility of a positive relationship between aspects of fitness and population size 50 years ago. Until recently, however, this concept was generally regarded as an intriguing but relatively unimportant aspect of population ecology. Increasing appreciation that Allee effects must be incorporated into models of population dynamics and habitat use, together with recent interest in the implications of sociality for conservation, have shown that for ecology and conservation the consequences of the Allee effect are profound. The Allee effect can be regarded not only as a suite of problems associated with rarity, but also as the basis of animal sociality. PMID- 10481205 TI - Inverse density dependence and the Allee effect. AB - The Allee effect describes a scenario in which populations at low numbers are affected by a positive relationship between population growth rate and density, which increases their likelihood of extinction. The importance of this dynamic process in ecology has been under-appreciated and recent evidence now suggests that it might have an impact on the population dynamics of many plant and animal species. Studies of the causal mechanisms generating Allee effects in small populations could provide a key to understanding their dynamics. PMID- 10481206 TI - The cuckoo's contested history. PMID- 10481208 TI - Introduction PMID- 10481207 TI - Cardioprotection with sodium-hydrogen exchange inhibition: An archetype of "Bench to bedside" PMID- 10481209 TI - Functional and cellular regulation of the myocardial Na+/H+ exchanger. AB - The Na(+)/H(+) exchanger is a pH-regulatory protein present in the plasma membrane of cardiomyocytes and other cell types. In response to intracellular acidosis, the protein removes one intracellular proton in exchange for an extracellular sodium. The protein consists of a membrane transporting domain and a regulatory cytosolic domain. The regulatory cytosolic domain mediates the stimulation of the membrane domain. Hormonal stimulation of myocardial cells results in activation of the antiporter, possibly through protein kinases and other regulatory proteins. Several hormones and growth factors have been shown to stimulate the antiporter in the myocardium, including endothelin, thrombin, angiotensin II, and alpha(1)-adrenergic stimulation. The exact mechanisms involved in this stimulation are as yet unclear, and may be important in regulation of the Na(+)/H(+) exchanger during ischemia and reperfusion. PMID- 10481210 TI - Sodium-hydrogen exchange and platelet function. AB - On stimulation of platelets with agonists, for example, thrombin, a rapid rise in intracellular pH is observed. This alkalinization is mediated by an increase in transport activity of the Na(+)/H(+) exchanger isoform NHE1. In addition to this Na(+)/H(+) exchange mechanism, platelets express bicarbonate/chloride exchangers, which also contribute to pH(i) homeostasis. The main functions of NHE1 in platelets include pH(i) control, volume regulation, and participation in cell signaling. The isoform NHE1 is highly sensitive toward inhibition by EIPA, Hoe694, and Hoe642. The regulation of NHE1 activity is complex and is not completely understood. It includes the MAP kinase cascade, the Ca/calmodulin system, several heterotrimeric G proteins (Galpha12, Galpha13, Galphaq, and Galphai), small G proteins (ras, cdc42, rhoA), and downstream kinases (e.g., p160ROCK). Volume challenges stimulate tyrosine phosphorylation of cytoplasmic proteins, which ultimately activate NHE1. Thrombin, thromboxane, platelet activating factor, angiotensin II, endothelin, phorbol ester, and Ca(2+) ionophors stimulate NHE1 activity in platelets. Blockade of platelet NHE1 can inhibit platelet activation. With the development of highly specific NHE1 inhibitors, detailed investigation of the relationships between NHE1 activity and platelet activation now becomes feasible. PMID- 10481212 TI - Mechanisms of protection of the ischemic and reperfused myocardium by sodium hydrogen exchange inhibition. AB - Sodium-hydrogen exchange (Na-H exchange) is a major regulator of intracellular pH and is one of the major mechanisms for restoring pH after ischemia-induced intracellular acidosis. However, activation of Na-H exchange during ischemia and reperfusion is also involved in paradoxical induction of cell injury. This likely reflects the fact that activation of the exchanger is closely coupled to sodium influx and, as a consequence, to elevation in intracellular calcium concentrations through sodium-calcium exchange. In addition to intracellular acidosis, other factors can also stimulate the exchanger, including various autocrine and paracrine factors, such as endothelin-1, angiotensin II, alpha(1) adrenergic agonists, as well as toxic agents, such as hydrogen peroxide and lysophosphatidylcholine. Although at least six Na-H exchange isoforms have thus far been identified, it appears that the 1 subtype, termed NHE1, is the predominant isoform in the mammalian myocardium. Effective pharmacological inhibitors of Na-H exchange, including those that are NHE1 specific, have been extensively demonstrated to protect the ischemic and reperfused myocardium in terms of improved systolic and diastolic function, preservation of cellular ultrastructure, attenuation of the incidence of arrhythmias, and reduction of apoptosis. Moreover, the salutary effects of these agents have been demonstrated using a variety of experimental models as well as animal species, suggesting that the role of Na-H exchange in mediating injury is not species specific. Thus, Na-H exchange represents an important target for pharmacological intervention in attenuation of ischemia and reperfusion-induced cardiac injury. Coupled with the low potential for toxicity of the agents, Na-H exchange inhibition could emerge as an effective therapeutic strategy in cardiac disorders, particularly involving conditions associated with ischemia and reperfusion. PMID- 10481211 TI - Regulation of cardiac sarcolemmal Na+/H+ exchanger activity: potential pathophysiological significance of endogenous mediators and oxidant stress. AB - The cardiac sarcolemmal Na(+)/H(+) exchanger (NHE) extrudes one H(+) in exchange for one Na(+) entering the myocyte, utilizing for its driving force the inwardly directed Na(+) gradient maintained by the Na(+), K(+)-ATPase. The exchanger is quiescent at physiological values of intracellular pH but becomes activated in response to intracellular acidosis. Recent evidence suggests that a variety of extracellular signals (e.g., adrenergic agonists, thrombin, endothelin, and oxidant stress) also modulate sarcolemmal NHE activity by altering its sensitivity to intracellular H(+). Because sarcolemmal NHE activity is believed to be an important determinant of the extent of myocardial injury during ischemia and reperfusion, regulation of exchanger activity by factors that are associated with ischemia is likely to be pathophysiological importance. PMID- 10481213 TI - If ischemic preconditioning is the gold standard, has a platinum standard of cardioprotection arrived? Comparison with NHE inhibition. PMID- 10481214 TI - Role of the sodium-hydrogen exchanger in ischemia-reperfusion injury in diabetes. AB - Ischemic heart disease is a significant problem in the diabetic population. Animal models of diabetes show a paradoxical resistance to ischemic challenge. The present treatise will discuss the mechanics involved and the central role that Na+-H+ exchanger plays in this response to ischemic-reperfusion injury. PMID- 10481217 TI - Reply from I.P.F. Owens, C. Rowe and A.L.R. Thomas. PMID- 10481216 TI - Development of the Na+/H+ exchange inhibitor cariporide as a cardioprotective drug: from the laboratory to the GUARDIAN trial. PMID- 10481218 TI - Resource selection functions: taking space seriously? PMID- 10481215 TI - Protection of the myocardium with sodium-hydrogen exchange inhibitors: A cardiac surgical perspective. AB - Strategies for myocardial protection in cardiac surgery are directed at the prevention of procedure-induced ischemia/reperfusion injury as well as metabolic resuscitation in acute ischemic syndromes. Postreperfusion myocardial dysfunction remains a significant clinical problem, most importantly in certain high-risk patient groups. The large body of experimental evidence demonstrating a significant role for sodium-hydrogen exchange activation in myocardial ischemia/reperfusion injury suggests that the ability to pharmacologically inhibit the exchanger presents a promising new approach to current myocardial preservation techniques. PMID- 10481219 TI - Actions of the pro-inflammatory cytokine IL-1 beta on central synaptic transmission. AB - It is now clear that IL-1 beta has profound and complex actions in the CNS. At pathophysiological concentrations it has an inhibitory effect on LTP in many regions of the hippocampus. There is also evidence emerging for a physiological role for IL-1 beta in the brain. Table 1 summarizes the effects of IL-1 beta and some other pro-inflammatory cytokines in the CNS. PMID- 10481220 TI - Development of the fluorescent microsphere technique for quantifying regional blood flow in small mammals. AB - We have demonstrated that the fluorescent microsphere technique can be used in small mammals for accurate determination of regional blood flows. In particular we have shown that 100% recovery of trapped microspheres is possible, that tissue digestion can be completed in a shorter time than previously reported, and the error-prone filtration method can be replaced with one of sedimentation. The method gave very good agreement among different fluorescent labels (r2 > 0.99) and low variability among tissues (mean coefficient of variation = 0.06). Simultaneous injection of radiolabelled and fluorescent microspheres established comparability between these methods (r2 = 0.96) for blood flows measured at rest, during vasodilator-induced hypotension, and in muscle hyperaemia during indirect electrical stimulation. Fluorescent microspheres can therefore replace radioactive microspheres for the determination of blood flow with advantages in both safety and cost, without loss of sensitivity. PMID- 10481221 TI - Non-muscarinic and non-nicotinic inhibition by the acetylcholine analogue carbachol of the delayed rectifier potassium current, iK, in rabbit isolated sino atrial node cells. AB - The effect of carbachol, an analogue of acetylcholine, on the delayed rectifier potassium current, iK, was investigated in rabbit isolated sino-atrial node cells using the whole cell patch clamp technique with amphotericin-permeabilized patches. In the presence of 500 nM atropine and 500 nM hexamethonium to block muscarinic and nicotinic receptors, respectively, 500 nM carbachol decreased the amplitude and rate of deactivation of iK without, however, affecting the slope of the iK activation curve. The same concentration of carbachol decreased the pacemaking rate of spontaneously active sino-atrial node cells by more than 13%. Thus, there is a non-muscarinic and non-nicotinic pathway for cholinergically induced reduction in the amplitude and rate of deactivation of iK that would appear to contribute to negative chronotropy in rabbit sinoatrial node pacemaker cells. PMID- 10481222 TI - Pyrimidine nucleotide-evoked inhibition of cyclic AMP accumulation in equine epithelial cells. AB - Uridine triphosphate (UTP) evoked inhibition of adrenaline-evoked cAMP accumulation in cultured equine epithelial cells (EC50, 1.8 +/- 0.2 microM) and this effect was mimicked by 5-Br-UTP (EC50, 6.6 +/- 1.8 microM) and uridine diphosphate (UDP; EC50, 96 +/- 26 microM). This inhibitory action of UTP was abolished by pre-treating cells with pertussis toxin (10 ng ml-1, 24 h). UTP (EC50, 2.3 +/- 0.3 microM) and 5-Br-UTP (EC50, 29.4 +/- 9.4 microM) also increased intracellular free calcium ([Ca2+]i) whilst UDP did not; the two effects are thus differentially sensitive to these pyrimidine nucleotides. ATP evoked cAMP accumulation in control cells and this response was unaffected by pertussis toxin. There is, therefore, no indication that ATP activates the pertussis toxin-sensitive inhibitory pathway. The UTP-evoked inhibition of cAMP accumulation was abolished by isobutylmethylxanthine (IBMX, 5 mM) and so the negative control over cAMP levels appears to be mediated by receptors that are selectively activated by pyrimidine nucleotides and permit control over phosphodiesterase activity. PMID- 10481223 TI - Energy metabolism of rat skeletal muscle modulated by the rate of perfusion flow. AB - In order to advance our understanding of the phenomenon of flow-induced increases in the metabolism of the relaxed muscle, the metabolic rate of the isolated rat gracilis muscle was investigated at 28 degrees C in vitro. The muscle was perfused with cell-free Krebs-Henseleit bicarbonate buffer containing 5% bovine serum albumin and 5 mM glucose, saturated with a gas mixture of 95% O2 and 5% CO2 and simultaneously superfused with a medium saturated with with a low O2 gas mixture (1% O2, 5% CO2 and 94% N2). Two different perfusion flow rates (0.054 and 0.100 ml min-1) have been used. Their influence on oxygen consumption and lactate production has been measured. After a 100 min perfusion period, the muscle was freeze-clamped and analysed for ATP, phosphocreatine, creatine, lactate, pyruvate, inorganic phosphate and glycogen content. The energy state of the cell and the proportions of glycolytic and mitochondrial fluxes of ATP synthesis were evaluated. During perfusion at the low flow rate of 0.054 ml min-1, the oxygen uptake was 45 +/- 9 nmol min-1 (g wet wt)-1, accompanied by a dominance of anaerobic glycolytic synthesis of ATP over mitochondrial ATP synthesis, even though the total delivery of oxygen to muscle was three times higher than oxygen consumption. Increasing the perfusion flow rate to 0.100 ml min-1 increased the oxygen uptake to 120 +/- 6 nmol min-1 (g wet wt)-1, thus leading to a prevalence of mitochondrial ATP synthesis over glycolytic ATP synthesis. The inner stores of glycogen served as the main substrate of energy metabolism and the role of exogenous substrates in the flow-stimulated increase of oxygen uptake was negligible. The increase in perfusion rate also enhanced the energy state of the muscle fibres, which was expressed either as the creatine charge or as the value of the change of Gibbs free energy of ATP hydrolysis. Data indicate that the change of perfusion flow rate per se, apart from oxygen and exogenous substrate supply, elicits changes in the regulation of energy metabolism within non contracting skeletal muscle under open microcirculation. PMID- 10481224 TI - The relaxant effects of parathyroid hormone(1-34) and parathyroid hormone-related protein(1-34) on ovine reticulo-ruminal smooth muscle in vivo. AB - The motility of the reticulo-rumen has been measured in trained, conscious sheep using inflated balloons temporarily introduced to selected regions of that forestomach. The frequency and amplitude of the contractions of the reticulum and both the A and B waves of contraction of the rumen were measured under the same conditions before, during and after the administration of an i.v. bolus of either parathyroid hormone (PTH(1-34)) or PTH-related protein (PTHrP(1-34)) followed by its i.v. infusion. These two peptides are known to share a common receptor in other organs, e.g. the kidney. In this study they both showed an inhibitory effect on reticulo-ruminal motility. The effect of PTHrP(1-34) on the rate of ruminal blood flow was also examined and a significant reduction observed, after a transient increase. The secretion of endogenous PTH(1-34) was stimulated by a 32% reduction in the plasma calcium ion concentration induced by an i.v. infusion of sodium citrate. Associated with this were significant reductions in reticulo ruminal motility, e.g. the reduction in the mean amplitude of the reticular contractions reflected the reduction in plasma calcium ion concentration. When the PTH(1-34)/PTHrP(1-34) receptor was blocked with [Asn10,Leu11,D-Trp12]PTHrP(7 34) before and during the induction of hypocalcaemia, all but one of the parameters of reticulo-ruminal motility were normalized. Indeed, by the day following the administration of this blocking agent, all these parameters had returned to their normal range. It is concluded that stimulation of the PTH(1 34)/PTHrP(1-34) receptor in reticulo-ruminal smooth muscle reduces the motility of this tissue and may play a role in the depression of motility of the digestive tract which is characteristic of clinical milk fever in the dairy cow. PMID- 10481225 TI - Effects of reducing submandibular blood flow on secretory responses to parasympathetic stimulation in anaesthetized cats. AB - Submandibular secretory responses to stimulation of the parasympathetic chorda lingual nerve were investigated in five anaesthetized cats before, during and after withdrawal of blood (ca 20 ml kg-1) in order to investigate the consequences of a reduced blood flow through the gland. Stimulation at different frequencies (2, 4, 6 and 8 Hz) evoked a frequency-dependent increase in the flow of submandibular saliva, sodium concentration, electrolyte and protein output. When the blood pressure was reduced (by about 50%) there was a significant reduction in submandibular blood flow and the secretion of both saliva and protein during stimulation. Under each set of conditions the flow of saliva was linearly related to the blood flow through the gland. It is concluded that submandibular secretory responses to electrical stimulation of the parasympathetic innervation can be significantly attenuated when the blood flow through the gland is reduced under the conditions employed in this study. PMID- 10481226 TI - Measurement of changes in glomerular filtration rate induced by atrial natriuretic peptide in the rat kidney. AB - This study was undertaken to improve the measurement of glomerular filtration rate (GFR) during the acute diuretic phase induced by atrial natriuretic peptide (ANP), which may indeed alter the renal clearance of inulin (GFRCL) due to dead space error. A technique to measure GFR without urine collections was therefore developed in anaesthetized rats prepared as for micropuncture. To do so, arterial blood was periodically collected and renal venous blood was withdrawn simultaneously from a catheter inserted into the left suprarenal vein to determine the renal extraction coefficient of inulin (CEIN). In addition, renal blood flow (RBF) was continuously measured with an electromagnetic flow transducer fitted around the left renal artery to estimate renal plasma flow (RPF). GFR (GFRCE) was then calculated as the product of RPF and CEIN. To study the effects of ANP on GFR, rats were injected i.v. with 10 microliters of saline without (n = 6; vehicle) or with 1 microgram ANP (n = 6; ANP) and GFRCE and GFRCL were compared before and after each treatment. They did not differ significantly during baseline measurements in each experimental group and were not modified after vehicle. Similarly, RBF remained constant. In contrast, RBF and GFRCE increased rapidly and simultaneously 90 s after ANP, from 9.07 +/- 0.25 to 10.07 +/- 0.35 (12%) and from 1.209 +/- 0.188 to 1.715 +/- 0.190 ml min-1 (42%), respectively (P < 0.05). GFRCL increased to an even greater extent (88%). Moreover, the peak enhancement of GFRCL was delayed and occurred 180 s after ANP. The renal clearance of inulin was thus unduly elevated due to sudden changes in the dead space induced by the diuretic effect of ANP. In conclusion, determination of glomerular filtration rate by the method of renal extraction of inulin provided more reliable results than those achieved using the classical method of renal clearance of inulin. Moreover, it was sufficiently sensitive to detect small and transient changes in GFR induced by the injection of 1 microgram ANP. PMID- 10481227 TI - Pancreatic alpha cell function in the fetal foal during late gestation. AB - Plasma glucagon concentrations were measured in chronically catheterized fetal ponies and their mothers between 260 days of gestation and term (approximately 335 days). Fetal alpha cell responses to arginine and variations in fetal glycaemia were also examined during late gestation. Immunoreactive glucagon was present in fetal plasma at 260 days of gestation and its concentration in utero increased after 320 days and then again at birth. Maternal plasma glucagon concentrations were higher after 300 days than earlier in gestation but were lower than the corresponding fetal value throughout the period of gestation studied. Fetal alpha cells responded rapidly to intravenous arginine infusion but not to changes in the fetal glucose level induced by maternal fasting for 36 h or by intrafetal infusion of glucose. The maximal increment in fetal plasma glucagon in response to arginine occurred at the end of the 5 min infusion and was positively correlated to the basal pre-infusion plasma glucagon concentrations. Fetal plasma glucagon concentrations were unaffected by either hyper- or hypoglycaemia. In contrast, maternal plasma glucagon levels were significantly increased by fasting. These observations indicate that equine pancreatic alpha cells are functional in utero but that they are unresponsive to variations in glycaemia until after birth. PMID- 10481228 TI - Effect of hypocalcaemia on glucose metabolism in hyperketonaemic piglets. AB - In nine 2- to 3-month-old hyperketonaemic piglets the kinetics of glucose and D beta-hydroxybutyrate (D-BHB) metabolism were studied during hypo- and normocalcaemia in paired experiments. Hyperketonaemia (1.3 and 2.5 mmol D-BHB (l plasma)-1) was generated by a stepwise increase of DL-BHB infusion. Hypocalcaemia spontaneously developed in five piglets due to an inherited calcitriol deficiency and was induced in four control piglets by a continuous infusion of Na2-EDTA. The method of single isotopic marker injections of glucose and D-BHB was used to calculate replacement rates, rate constants and half-lives of glucose and D-BHB in plasma. When DL-BHB was infused at the same rate into normo- and hypocalcaemic piglets, hypocalcaemia reduced the rate constant of glucose by 20-30% and the replacement rate of glucose by 34%. In the presence of hyperketonaemia, hypocalcaemia increased the rate of replacement of D-BHB by 6-40%. The replacement rate represents the sum of endogenous production and the rate of DL BHB infusion. This observation shows that the endogenous production of D-BHB was higher during hypocalcaemia than during normocalcaemia. PMID- 10481229 TI - The effects of in vivo pulmonary oxygenation on lung liquid production in near term fetal sheep. AB - Lung liquid (LL) is secreted into the fetal lung lumen, but it must be rapidly absorbed at birth to allow air breathing. In vitro studies have implicated oxygen as a possible factor causing the switch from secretion to absorption of lung liquid at birth. We developed a technique of oxygenating the fetal lung using liquid ventilation with haemoglobin (Hb) solutions in chronically catheterized fetal lambs (129-140 days gestation; term, 147 days). In some experiments 2,3 diphosphoglycerate (DPG) was added to increase oxygen delivery. LL secretion rate (Jv) was measured using an indicator dilution method. Eighteen fetuses were divided into four groups and ventilated with liquid under the following conditions: (i) Hb with oxygen, (ii) Hb without oxygen, (iii) Hb with DPG and oxygen and (iv) Hb and DPG without oxygen. There was a significant rise (2.6 mmHg, P < 0.02) in fetal arterial Po2 in group iii, but in none of the other groups. In the first 3 h of liquid ventilation there was no difference in Jv between the groups. In group i, during hours 4-6 of liquid ventilation, there was a significant rise in secretion rate from 2.25 +/- 0.88 to 3.74 +/- 0.85 ml h-1 kg-1 (P < 0.001). In group iii, when comparing Jv in the first 3 h of liquid ventilation with that in the following 3 h period of liquid ventilation, a strong trend towards reduction in secretion was observed, falling from 3.03 +/- 0.65 to 0.74 +/- 0.92 ml h-1 kg-1 (three of the four experiments showed a significant decrease in Jv in hours 4-6). These experiments indicate that oxygen delivered to the fetus using liquid ventilation with haemoglobin solutions leads to increased LL secretion when oxygen delivery is small, and suggest there is a decrease in secretion with greater oxygen delivery to the lung. PMID- 10481230 TI - Laryngeal movements during the respiratory cycle measured with an endoscopic imaging technique in the conscious horse at rest. AB - A video-laryngoscopic method, implemented with an algorithm for the correction of the deformation inherent in the endoscope optical system, has been used to measure the dorsoventral diameter (Drg) and the cross-sectional area (CSArg) of the rima glottidis in five healthy workhorses during conscious breathing at rest. Simultaneous recording of the respiratory airflow was also obtained in two horses. Drg measured 82.7 +/- 4.5 mm (mean +/- S.D.) independently of the respiratory phase, and did not differ from the measurement in post-mortem anatomical specimens of the same horses. CSArg ranged from 1130 +/- 117 mm2 (mean +/- S.D.) during the inspiratory phase to 640 +/- 242 mm2 during the expiratory phase; being always narrower than tracheal cross-sectional area, which was 1616 +/- 224 mm2, as determined from anatomical specimens. Both inspiratory and expiratory airflow waves displayed a biphasic pattern. Maximal laryngeal opening occurred in phase with the second inspiratory peak, while during expiration CSArg attained a minimum value during the first expiratory peak which was significantly smaller (P < 0.01) than the area subsequently maintained during the rest of the expiratiory phase. These quantitative measurements of equine laryngeal movements substantiate the important role played by the larynx in regulating upper airway respiratory resistance and the expiratory airflow pattern at rest. PMID- 10481231 TI - Oxygen uptake kinetics of older humans are slowed with age but are unaffected by hyperoxia. AB - Cross-sectional studies have compared the oxygen uptake (VO2) kinetics during the on-transient of moderate intensity exercise in older and younger adults. The slower values in the older adults may have been due to an age-related reduction in the capacity for O2 transport or alternatively a reduced intramuscular oxidative capacity. We studied: (1) the effects of ageing on VO2 kinetics in older adults on two occasions 9 years apart, and (2) the effect of hyperoxia on VO2 kinetics at the second test time. After a 9 year period, follow-up testing was undertaken on seven older adults (78 +/- 5 years, mean +/- S.D.). They each performed six repeats of 6 min bouts of constant-load cycle exercise from loadless cycling to 80% of their ventilatory threshold. They breathed one of two gas mixtures (euoxia: inspired O2 fraction, FI,O2, 0.21; hyperoxia: FI,O2, 0.70) on different trials determined on a random basis. Breath-by-breath VO2 data were time aligned and ensemble averaged. VO2 kinetics, modelled with a single exponential from phase 2 onset (+20 s) to steady state and described by the exponential time constant (tau) were compared with data collected from the same adults 9 years earlier. One-way repeated measures analysis of variance revealed that tau was slowed significantly with age (from 30 +/- 8 to 46 +/- 10 s), but was unaffected by hyperoxia (43 +/- 15 s). We concluded that: (1) in older adults studied longitudinally over a 9 year period, the on-transient VO2 kinetics are slowed, in agreement with, but to a greater extent, than from cross-sectional data; and (2) the phase 2 time constant (tau) for these older adults was not accelerated by hyperoxic breathing. Thus the expected hyperoxia-induced increase in the capacity for O2 transport was not associated with faster on-transient VO2 kinetics suggesting either that O2 transport may not limit VO2 kinetics during the 8th decade, or that O2 transport was not improved with hyperoxia. PMID- 10481232 TI - The effects of caffeine on the kinetics of O2 uptake, CO2 production and expiratory ventilation in humans during the on-transient of moderate and heavy intensity exercise. AB - In order to test the hypothesis that glycogen sparing observed early during exercise following caffeine ingestion was a consequence of tighter metabolic control reflected in faster VO2 kinetics, we examined the effect of caffeine ingestion on oxygen uptake (VO2), carbon dioxide production (VCO2) and expiratory ventilation (VE) kinetics at the onset of both moderate (MOD) and heavy (HVY) intensity exercise. Male subjects (n = 10) were assigned to either a MOD (50% VO2,max, n = 5) or HVY (80% VO2,max, n = 5) exercise condition. Constant-load cycle ergometer exercise was performed as a step function from loadless cycling 1 h after ingestion of either dextrose (placebo, PLAC) or caffeine (CAFF; 6 mg (kg body mass)-1). Alveolar gas exchange was measured breath-by-breath. A 2- or 3 component exponential model, fitted through the entire exercise transient, was used to analyse gas exchange and ventilatory data for the determination of total lag time (TLT: the time taken to attain 63% of the total exponential increase). Caffeine had no effect on TLT for VO2 kinetics at either exercise intensity (MOD: 36 +/- 14 s (PLAC) and 41 +/- 10 s (CAFF); HVY: 99 +/- 30 s (PLAC) and 103 +/- 26 (CAFF) (mean +/- S.D.)). TLT for VE was increased with caffeine at both exercise intensities (MOD: 50 +/- 20 s (PLAC) and 59 +/- 21 s (CAFF); HVY: 168 +/- 35 s (PLAC) and 203 +/- 48 s (CAFF)) and for VCO2 during MOD only (MOD: 47 +/- 14 s (PLAC) and 53 +/- 17 s (CAFF); HVY: 65 +/- 13 s (PLAC) and 69 +/- 17 s (CAFF)). Contrary to our hypothesis, the metabolic effects of caffeine did not alter the on-transient VO2 kinetics in moderate or heavy exercise. VCO2 kinetics were slowed by a reduction in CO2 stores reflected in pre-exercise and exercise endtidal CO2 pressure (PET,CO2) and plasma PCO2 which, we propose, contributed to slowed VE kinetics. PMID- 10481233 TI - Effect of temperature on muscle metabolism during submaximal exercise in humans. AB - To study the effect of temperature on muscle metabolism during submaximal exercise, six endurance-trained men had one thigh warmed and the other cooled for 40 min prior to exercise using water-perfused cuffs. One cuff was perfused with water at 50-55 degrees C (HL) with the other being perfused with water at 0 degree C (CL). With the cuffs still in position, subjects performed cycling exercise for 20 min at a work load corresponding to 70% VO2,peak (where VO2,peak is peak pulmonary oxygen uptake) in comfortable ambient conditions (20-22 degrees C). Muscle biopsies were obtained prior to and following exercise and forearm venous blood was collected prior to and throughout the exercise period. Muscle temperature (Tmus) was not different prior to treatment, but treatment resulted in a large difference in pre-exercise Tmus (difference = 6.9 +/- 0.9 degrees C; P < 0.01). Although this difference was reduced following exercise; it was nonetheless significant (difference = 0.4 +/- 0.1 degree C; P < 0.05). Intramuscular [ATP] was not affected by either exercise or muscle temperature. [Phosphocreatine] decreased (P < 0.01) and [creatine] increased (P < 0.01) with exercise but were not different when comparing HL with CL. Muscle lactate concentration was not different prior to treatment nor following exercise when comparing HL with CL. Muscle glycogen concentration was not different when comparing the trials before treatment, but the post-exercise value was lower (P < 0.05) in HL compared with CL. Thus, net muscle glycogen use was greater during exercise with heating (208 +/- 23 vs. 118 +/- 22 mmol kg-1 for HL and CL, respectively; P < 0.05). These data demonstrate that muscle glycogen use is augmented by increases in intramuscular temperature despite no differences in high energy phosphagen metabolism being observed when comparing treatments. This suggests that the increase in carbohydrate utilization occurred as a direct effect of an elevated muscle temperature and was not secondary to allosteric activation of enzymes mediated by a reduced ATP content. PMID- 10481234 TI - Candidate genes involved in vascular matrix remodelling in atherogenesis. PMID- 10481235 TI - Molecular modelling of the biosynthesis of the RNA-editing enzyme APOBEC-1, responsible for generating the alternative forms of apolipoprotein B. AB - We discovered in 1987 that the shorter form of apolipoprotein B (B48) synthesized in the intestine is due to the action, previously unrecognized in mammalian cells, of an mRNA-editing process, and more recently we demonstrated that this was due to a specific enzyme (APOBEC-1) with cytidine deaminase activity. We show here, by sequence alignment, molecular modelling and mutagenesis, that APOBEC-1 is a cytidine deaminase, responsible for editing apoB mRNA, and that is related in crystal structure to the cytidine deaminase of Escherichia coli (ECCDA). The two enzymes are both homodimers with composite active sites formed with loops from each monomer. In the sequence of APOBEC-1, three gaps compared with ECCDA match the size and contour of the minimal RNA substrate. We propose a model in which the asymmetric binding of one active site to the substrate cytidine which is positioned by the downstream binding of the product uridine and that this helps to target the other active site for deamination. PMID- 10481236 TI - Speeds of actin translocation in vitro by myosins extracted from single rat muscle fibres of different types. AB - As skeletal muscle fibres mostly express a single myosin isoform, they are a potential source of pure myosin isoforms. A technique is described that allows extraction and identification of pure myosin isoforms from single fibres, and testing of such myosins in an in vitro motility assay (IVMA). The results show that the extraction procedure does not alter myosin function and support the view that single fibres are reliable sources of purified myosin isoforms for IVMA. PMID- 10481237 TI - Tremulousness--the perception of tremor in man. AB - The thresholds for detection of a tremor-like movement are described. Detection is poorest at frequencies which correspond to natural tremor. The results are in accord with the failure of persons often to notice that their hands are tremulous. PMID- 10481238 TI - Sensitivity of vertical jumping performance to changes in muscle stimulation onset times: a simulation study. AB - The effect of muscle stimulation dynamics on the sensitivity of jumping achievement to variations in timing of muscle stimulation onsets was investigated. Vertical squat jumps were simulated using a forward dynamic model of the human musculoskeletal system. The model calculates the motion of body segments corresponding to STIM(t) of six major muscle groups of the lower extremity, where STIM is muscle stimulation level. For each muscle, STIM was allowed to switch "on" only once. The subsequent rise of STIM to its maximum was described using a sigmoidal curve, the dynamics of which was expressed as rise time (RT). For different values of stimulation RT, the optimal set of onset times was determined using dynamic optimization with height reached by the center of mass as performance criterion. Subsequently, 200 jumps were simulated in which the optimal muscle stimulation onset times were perturbed by adding to each a small number taken from a Gaussian-distributed set of pseudo-random numbers. The distribution of heights achieved in these perturbed jumps was used to quantify the sensitivity of jump height to variations in timing of muscle stimulation onsets. It was found that with increasing RT, the sensitivity of jump height to timing of stimulation onset times decreased. To try and understand this finding, a post-hoc analysis was performed on the perturbed jumps. Jump height was most sensitive to errors in the delay between stimulation onset times of proximal muscles and stimulation onset times of plantar flexors. It is explained how errors in this delay cause aberrations in the configuration of the system, which are regenerative and lead to relatively large jump height deficits. With increasing RT, the initial aberrations due to erroneous timing of muscle stimulation are smaller, and the regeneration is less pronounced. Finally, it is speculated that human subjects decrease or increase RT depending on the relative importance of different performance criteria. PMID- 10481239 TI - A linear canal-otolith interaction model to describe the human vestibulo-ocular reflex. AB - A control systems model of the vestibulo-ocular reflex (VOR) originally derived for yaw rotation about an eccentric axis (Crane et al. 1997) was applied to data collected during ambulation and dynamic posturography. The model incorporates a linear summation of an otolith response due to head translation scaled by target distance, adding to a semi-circular canal response that depends only on angular head rotation. The results of the model were compared with human experimental data by supplying head angular velocity as determined by magnetic search coil recording as the input for the canal branch of the model and supplying linear acceleration as determined by flux gate magnetometer measurements of otolith position. The model was fit to data by determining otolith weighting that enabled the model to best fit the data. We fit to the model experimental data from normal subjects who were: standing quietly, walking, running, or making active sinusoidal head movements. We also fit data obtained during dynamic posturography tasks of: standing on a platform sliding in a horizontal plane at 0.2 Hz, standing directly on a platform tilting at 0.1 Hz, and standing on the tilting platform buffered by a 5-cm thick foam rubber cushion. Each task was done with the subject attending a target approximately 500, 100, or 50 cm distant, both in light and darkness. The model accurately predicted the observed VOR response during each test. Greater otolith weighting was required for near targets for nearly all activities, consistent with weights for the otolith component found in previous studies employing imposed rotations. The only exceptions were for vertical axis motion during standing, sliding, and tilting when the platform was buffered with foam rubber. In the horizontal axis, the model always fit near target data better with a higher otolith component. Otolith weights were similar with the target visible and in darkness. The model predicts eye movement during both passive whole-body rotation and free head movement in space implying that the VOR is controlled by a similar mechanism during both situations. Factors such as vision, proprioception, and efference copy that are available during head free motion but not during whole-body rotation are probably not important to gaze stabilization during ambulation and postural stabilizing movement. The linearity of the canal-otolith interaction was tested by re-analysis of the whole body rotation data on which the model is based (Crane et al. 1997). Normalized otolith mediated gain enhancement was determined for each axis of rotation. This analysis uncovered minor non-linearities in the canal-otolith interaction at frequencies above 1.6 Hz and when the axis of rotation was posterior to the head. PMID- 10481240 TI - Conditional entropy approach for the evaluation of the coupling strength. AB - A method that enables measurement of the degree of coupling between two signals is presented. The method is based on the definition of an uncoupling function calculating, by means of entropy rates, the minimum amount of independent information (i.e. the information carried by one signal which cannot be derived from the other). An estimator of the uncoupling function able to deal with short segments of data (a few hundred samples) is proposed, thus enabling the method to be used for usual experimental recordings. A synchronisation index is derived from the estimate of the uncoupling function by means of a minimisation procedure. It quantifies the maximum amount of information exchanged between the two signals. Simulations in which non-linear coordination schemes are produced and changes in the coupling strength are artificially induced are used to check the ability of the proposed index to measure the degree of synchronisation between signals. The synchronisation analysis is utilised to measure the coupling strength between the beat-to-beat variability of the sympathetic discharge and ventilation in decerebrate artificially ventilated cats and the degree of synchronisation between the beat-to-beat variability of the heart period and ventricular repolarisation interval in normal subjects and myocardial infarction patients. The sympathetic discharge and ventilation are strongly coupled and the coupling strength is not affected by manoeuvres capable of increasing or depressing sympathetic activity. The synchronisation is lost after spinalisation. The synchronisation analysis confirms that the heart period and ventricular repolarisation interval are well coordinated. In normal subjects, the synchronisation index is not modified by experimental conditions inducing changes in the sympathovagal balance. On the contrary, it strongly decreases after myocardial infarction, thus detecting and measuring the uncoupling between the heart period and ventricular repolarisation interval. PMID- 10481241 TI - Simulation of EEG: dynamic changes in synaptic efficacy, cerebral rhythms, and dissipative and generative activity in cortex. AB - A simulation of electrocortical activity based upon coupled local aggregates of excitatory and inhibitory cells was modified to include rapid dynamic variations of synaptic efficacy attributable to reversal potentials and related effects. The modified simulation reproduces the rhythmic phenomena observed in real EEG, including the theta, alpha, beta and gamma rhythms, in association with physiologically realistic pulse densities. At high levels of cortical activation, generative activity with a 40-Hz center frequency emerges, suggesting a basis for the occurrence of phase changes and "edge of chaos" dynamics. These local oscillation properties complement the dissipative travelling wave and synchronous oscillation effects attributable to longer range excitatory couplings, as previously demonstrated in related simulations. Results of variation of parameters provide a first approximation to the anticipated effects of slow physiological time variations in gains and lags, and some predictions of the model are described. PMID- 10481242 TI - Analytical description of the evolution of neural networks: learning rules and complexity. AB - We present the foundation of a physical formalism that allows us to characterize the dynamics of the evolution of neural networks both in regard to the network configuration and to network performance. Model runs were performed on a simple network consisting of six neurons, allowing complete analytical description of the network's behaviour. Order parameters are characterized that allow an analytical description of critical periods in network evolution. Thus, correlations of the local dynamics and the system's global behaviour could be computed. It is shown that local learning rules are sufficient to model complex dynamical aspects of the evolution of networks. It is demonstrated in how far novel statistical formalisms, e.g. neural complexity, can be employed to evaluate the system's dynamics. The introduction of order parameters allows an analytical characterization of transient phases in the network's behaviour, correlating network connectivity with neuronal firing patterns. The relevance of this approach for the interpretation of physiological data is discussed. PMID- 10481243 TI - Effects of culture parameters on the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenol (2,4-DCP) by selected fungi. Groupe pour l'Etude du Devenir des Xenobiotiques dans l'Environnement (GEDEXE). AB - In order to enhance 2,4-D and 2,4-DCP degradation by four selected fungi (Cunninghamella elegans, C. echinulata, Rhizoctonia solani and Verticillium lecanii), three culture parameters (initial chemical concentration, amounts of glucose and nitrogen) were varied. The levels of both xenobiotics in the culture media were monitored by HPLC analysis after five days of cultivation. The best results were obtained at low initial concentration (20 mg.L-1 vs 100) and with low amounts of glucose (5 g.L-1 vs 10) and nitrogen (2.4 mM vs 24). When these two elements were lacking from the culture media, biodegradation was not suppressed, but took place to a lesser extent. Thus, initial chemical concentration and amounts of carbon and nitrogen, in the culture medium, were shown to strongly influence the extent of 2,4-D and 2,4-DCP removal by fungi. PMID- 10481244 TI - Monitoring the toxicity of phenolic chemicals to activated sludge using a novel optical scanning respirometer. AB - This paper reports the development of optical scanning respirometer for determining the toxicity of chemicals to activated sludge. The respirometer is used to measure the dissolved oxygen concentration by monitoring the luminescence intensity of ruthenium dye immobilized in a polymer film in contact with the wastewater sample. The intensity is related to the extent of oxygen quenching of luminescence. The toxicity of chemicals can be evaluated by measuring the inhibition on respiration rate of microorganism using the scanning respirometer. The IC50 values (concentration of a chemical that exhibits 50% respiration inhibition) of various phenolic chemicals in activated sludge were determined. The performance of this method is compared with other toxicity methods. The experimental results indicate that the reproducibility and sensitivity of this respirometer are reasonably good. PMID- 10481245 TI - Estimating the ozone formation potential of volatile aromatic compounds in vehicle tunnels. AB - Two vehicle tunnels located in the Taipei area were selected to study the ozone formation potential of volatile aromatic compounds (VACs). Air samples in the two tunnels were collected using canisters. Analysis of VACs was conducted with a gas chromatograph equipped with a mass spectrometer. The total VOCs concentrations in Zefun tunnel ranged from 483.5 to 1032.2 (micrograms/m3) which increased with the increase in traffic volume. In the Lishin tunnel, the total VACs concentration ranged from 356.6 to 1961.3 (micrograms/m3) which was not well correlated with the traffic volume. The most predominant VACs in these vehicle tunnels were toluene and xylenes. Although the traffic volume and types of vehicles were not exactly the same, the characteristic ratios of the VACs concentrations were found to be similar. These results indicated that the existence of a specific characteristic ratio of VAC concentration was affected by the mobile sources. The maximum ozone formation potential resulted from the vehicles in the Lishin motorcycle tunnel which had higher values than the Zefun tunnel and much higher than the SPECIATE databases in the US via weighting the maximum incremental reactivity (MIR) scale. PMID- 10481246 TI - Biodegradation of coplanar polychlorinated biphenyls by anaerobic microorganisms from estuarine sediments. AB - In this study, we investigated the biodegradability of biphenyl and 5 congeners (one non-planar and four coplanar) of polychlorinated biphenyl (PCB). Biphenyl, the non-planar congener 2,3',4',5-tetrachlorobiphenyl (25-34 CB), and the four coplanar congeners 3,3',4,4'-tetrachlorobiphenyl (34-34 CB), 3,4,4',5 tetrachlorobiphenyl (345-4 CB), 3,3',4,4',5-pentachlorobiphenyl (345-34 CB), and 3,3',4,4',5,5'-hexachlorobiphenyl (345-345 CB) were amended at a concentration of 10 mg/L into anoxic sediment slurries collected from the estuaries of the Tansui River and the Erjen River. During 2 years' incubation under sulfidogenic conditions, biphenyl was persistent, while all other chlorinated congeners, except for 345-345 CB, were dechlorinated with or without a lag period in sediment slurries collected from both rivers. Dechlorination of coplanar and non planar congeners began with para chlorine removal. All para chlorines from the mono-, di-, and trichlorobiphenyl groups could be removed by sediment slurries from both rivers. Microbial communities in sediment from the Erjen River additionally fostered meta-dechlorination activity, but only after removal of all the para chlorines. Addition of Tween 20 (0.05%, v/v) into sediment slurries from the Tansui River did not enhance dechlorination rates or extents, but the addition of toluene- or 3-chlorobenzoate-adapted sediments enhanced dechlorination of 34-34 CB and 345-4 CB. PMID- 10481247 TI - Effects of elevated ozone concentration on the degradation of dichlorprop in soil. AB - An aerobic degradation study was conducted to estimate possible effects of elevated ozone concentration in air on the behaviour of dichlorprop. An average ozone concentration of 80 nL L-1 was chosen, which often occurs close to congested areas during late spring and summer. A control soil and an ozone exposed soil were kept under same conditions such as temperature, air flow and soil humidity. The use of 14C-labelled dichlorprop allowed to examine the fate of dichlorprop and follow the degradation products in soil. Exhaustive extraction of both soils yielded several fractions containing dichlorprop residues. Half lives of dichlorprop of both treatments were 5 days. After 32 days most of the residues in soil remained in the non extractable fraction. The elevated ozone concentration showed no significant effects on the degradation behaviour of dichlorprop and its metabolites but significant differences were found for the behaviour of the nonextractable residues and of the release of carbon dioxide, which were higher for control soil in comparison to the ozone variant. These findings suggest that even moderately elevated ozone concentration in air can effect mineralisation and fixation processes of dichlorprop. PMID- 10481248 TI - Methods for the analysis of PCBs in human food, faeces and serum. AB - A method was developed to determine trace concentrations of a range of individual PCB congeners in biological samples (serum, food and faeces) using GC-MS, to prepare a mass balance of PCBs in humans. A simple method for the analysis of PCBs in human serum, which excluded an extraction step, was first employed. Results indicated that the recoveries of 13C12 PCB spikes were variable. A soxhlet extraction step was added and was found to be efficient and reproducible. A quality control routine and method validation results are presented. In batch tests of the methods presented it was found that the serum analysis method gave within batch mean 13C12 spike recoveries of 98-120% and standard deviations between 6 and 20%. The food/faeces analysis method gave within-batch mean 13C12 spike recoveries of 88-100%, and within batch standard deviations between 4 and 12%. The batch to batch mean recovery for serum analysis was 100%, with an RSD of 9% for high spikes and 10% for low spikes. For food/faeces analysis the batch to batch average recovery was 110%, with an RSD of 5% for high spikes and 9% for low spikes. PMID- 10481249 TI - Sample preparation for GC analysis of selected pesticides in surface water. AB - A new isolation procedure for the determination of nitrogen/phosphorous containing pesticides and organochlorine pesticides in water was tested, and statistical evaluation of the recoveries was performed. The procedure, designed specifically for the analysis of semi-volatile compounds in water containing Suspended Particulate Matter (SPM), utilizes a specially designed filtration vessel coupled directly to an SPE cartridge. The studies were based on surface water samples (from the Vistula River) spiked with pesticides. SPM separation and analyte isolation/concentration were carried out in a special filtration vessel. Pesticides were sorbed both on the SPE cartridge and on the suspended matter. The cartridge and the filter with the suspended matter were extracted separately with a solvent, which enabled the determination of analyte distribution between the two. For organochlorine pesticides, between 3 and 60% of the initial amount was found on the filter, while the recoveries in the filtrate ranged from 30 to 98%. Total recoveries of organochlorine pesticides from surface water samples spiked with pesticides using the method described were high, ranging from 90 to 101%. The amounts of nitrogen/phosphorus containing pesticides recovered from the filter were lower than 2%, while those from the filtrate ranged from 69 to 92%. Total recoveries of nitrogen/phosphorus containing pesticides from surface water samples were high, ranging from 71 to 92%. PMID- 10481250 TI - Sorption of pentachlorophenol on peat-bentonite mixtures. AB - Batch kinetic and isotherm studies were carried out to determine the adsorptive characteristics of peat and bentonite mixtures for pentachlorophenol, and to examine the hydraulic conductivity of peat-bentonite mixtures to determine if they are applicable for use as cutoff barriers. Batch kinetic studies showed that over 90% of PCP was removed from water spiked with approximately 1 mg/l of PCP using a peat-bentonite (5%) mixture. The equilibrium time was 8 hours. The optimum pH range for adsorption of PCP by the peat-bentonite mixture was found to be 3-3.5. Batch isotherm studies showed that the adsorption of PCP by the peat bentonite mixture from aqueous solution was best described by the Freundlich isotherm equation. Batch adsorption studies using various ratios of bentonite in the mixture showed that the adsorption of PCP decreased linearly with increased amount of bentonite in the mixture, indicating that adsorption of PCP by the peat moss portion of the mixture was the dominant process. The inverse of the hydraulic conductivity was found to increase exponentially with an increase in the bentonite content of the mixture over the range studied. The minimum hydraulic conductivity observed was 3.3 x 10(-7) cm/s for a 50% peat-50% bentonite mixture. Peat-bentonite mixtures can be used to successfully remove PCP from aqueous media and can be used effectively as a barrier to attenuate the migration of PCP through soil and groundwater systems. PMID- 10481251 TI - A non-absorbable dietary fat substitute enhances elimination of persistent lipophilic contaminants in humans. AB - For individuals contaminated with persistent lipophilic pollutants, there is an urgent need for a therapy to enhance contaminant elimination from the body and hence reduce long term exposure. This study investigated the possibility of enhancing the excretion of native chemical via the faeces by augmenting the lipophilic properties of the faeces with the non-absorbable lipid substitute olestra. The faecal excretion of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), polychlorinated biphenyls (PCBs), and hexachlorobenzene (HCB) was measured in 3 volunteers. The excretion while eating an olestra-free diet was compared with the excretion while eating a diet supplemented wit 25 g/d of olestra. The excretion while on the olestra diet was higher by a factor of 1.5 11, depending on the compound. This resulted from higher concentrations of the contaminants in the faeces and higher excretion of faeces dry mass due to the food additive. Using 2,3,7,8-Cl4DD as an example, it was estimated that ingestion of 25 g/d of olestra would more than double the overall rate of elimination of this compound from the body. It is concluded that regular consumption of olestra may provide a therapeutic approach for reducing the body burden of persistent lipophilic contaminants. PMID- 10481252 TI - Metabolism and body-burden of PCBs in lactating dairy cows. AB - This paper uses data from a detailed 4 month PCB mass balance study on lactating cows fed a naturally contaminated background diet to quantify the metabolism and body-burdens of a range of PCB congeners. Dietary intake fluxes and milk and faecal output fluxes reported previously are combined with subcutaneous fat and blood PCB concentrations and with data from tissue/organ samples from a slaughtered animal to estimate the degree of metabolism within the cow. A total body burden is derived, showing that fat deposits account for approximately 98% of total PCB present in the lactating cow. The daily intake through feed (ng day 1) accounted for between 0.9 and 1.5% of the total body burden for persistent congeners (e.g. PCB 153) and up to 43% for those congeners which are readily metabolised (e.g. PCB 52). Detailed balances for a range of tri- to octa chlorinated PCBs are presented and clearly demonstrate that many congeners are metabolised effectively by lactating dairy cows (e.g. PCB 149) whilst others are efficiently transferred to human dairy food products. An approach to predicting the degree of metabolism for individual PCB congeners is presented which gives good agreement with observations. PMID- 10481253 TI - Toxicity evaluation of organic sediment extracts resolved by size exclusion chromatography using rainbow trout hepatocytes. AB - The (geno)toxicity of sediment dichloromethane extracts and fractions obtained by size exclusion chromatography were evaluated to investigate effects based on size fractionation. In this study, three sediments were selected according to their incremental contamination in PAHs and in PCBs: Hamilton harbour, Toronto bay and lake St. Clair sediments. Heavy metals, total sulfur and elemental sulfur (S8) were also determined in the (un)fractionated sediment extracts. The liver cells were exposed to concentrations of sediment extracts and fractionated samples for 24 h at 15 degrees C, afterwhich cell viability, cytochrome P4501A1 activity, available free Zn, DNA damage and oxidative stress were determined. The results showed that the sediment extracts contained high levels of sulfur most of which was found in the low molecular weight (LMW) region, i.e., the 2000-50 atomic mass unit (amu) fraction. Elemental sulfur (S8) accounted for 14-41% of extractable sulfur and were found to elute in the post-column volume (PCV) fraction despite its molecular weight of 256 amu. Heavy metals were found mainly in the HMW (i.e. the > 2000 amu) fraction and LMW fractions and very few or none were observed in the PCV fractions. In sediment extracts, sublethal effects were present principally by the HMW and LMW fractions suggesting that some chemicals were also associated with high molecular weight compounds of extractable organic matter. Less toxicity or effect was sometimes found in the extract indicating an antagonistic effect of the contaminants. We found that cell viability and genotoxicity evaluations could be performed on the unfractionated extracts while EROD, available Zn and oxidative stress measurements should be performed on the LMW fractions because of possible antagonist or shielding effects. Considering the cytotoxic responses, the best toxicity ranking in respect to contaminant levels in sediment extract was obtained with the LMW and PCV fractions which accounted for most of the toxic responses in the chromatographic fractions. Moreover, the shielding effect could be explained, in part, by the association of LMW contaminants to large macromolecules. PMID- 10481254 TI - Correlations of plasma growth hormone with somatostatin, gonadal steroid hormones and thyroid hormones in rainbow trout during sexual recrudescence. AB - The study explores the interrelationships among growth hormone (GH), somatostatin 14 (SRIF), non-esterified fatty acids (NEFA), gonadal steroid hormones and thyroid hormones (THs) in sexually recrudescent rainbow trout (Oncorhynchus mykiss) to examine aspects of the complex set of physiological changes associated with gonadal growth and maturation. Females exhibited significant decreases in plasma SRIF, NEFA and triiodo-L-thyronine (T3) concentrations, and a significant increase in plasma GH concentration associated with gonadal maturation, whereas in males, only SRIF and NEFA concentrations showed significant changes during testicular maturation. The declining SRIF levels during gonadal recrudescence may indicate a role for the hormone in the energy repartitioning processes that occur in both sexes at this time. Correlation analysis of plasma variables revealed a direct correlations between plasma NEFA and 17 beta-estradiol (E2) in females, an inverse correlation between NEFA and testosterone (T) in males, inverse correlations between GH and SRIF in both males and females, and inverse correlations between THs and SRIF concentrations in females. These marked gender differences in correlations likely reflect the different physiological challenges faced by the two sexes and emphasizes the need to consider gender, as well as maturity when studying the interactions of hormones. PMID- 10481255 TI - Mannose 6-phosphate receptors (MPR 300 and MPR 46) from a teleostean fish (trout). AB - Mannose 6-phosphate receptors (MPRs) are known to occur in mammals, birds, reptiles and amphibians. Here we provide evidence for the presence of two MPRs in fish, the earliest vertebrates. Using phosphomannan-Sepharose affinity chromatography, MPR 300 was purified from liver membrane extract of trout. The purified trout liver MPR 300 showed similar electrophoretic mobility as the goat liver receptor and a pH optimum of 7.0 for binding to phosphomannan. The presence of MPR 46 in fish was shown by metabolically labelling embryonic fish cells (Xiphophorus) and immunoprecipitation with an antibody against the cytoplasmic tail of human MPR 46 (anti-MSC1). This antibody had recently been shown to immunoprecipitate MPR 46 also from reptiles and amphibians. PMID- 10481256 TI - Reduced epinephrine-stimulated lipolytic activity in male golden-mantled ground squirrel during hibernation: an in situ microdialysis study. AB - We studied lipolytic activities in vivo in golden mantle ground squirrels during pre-hibernation and hibernation using microdialysis technique. Microdialysis probes were inserted into the abdominal subcutaneous adipose tissues. Baseline lipolysis were assessed by measuring glycerol concentration. Epinephrine stimulated lipolysis was also examined. Eight squirrels (four male, four female) were studied in each of the two stages. Basal glycerol concentrations were lower in the hibernating state than in the pre-hibernation state in male squirrels (P < 0.05). Epinephrine application induced glycerol release in male and female squirrels (P < 0.001) in both stages. Male squirrels demonstrated a reduced epinephrine-stimulated glycerol release in the hibernating state, which was not observed in female squirrels. PMID- 10481257 TI - Purification and characterization of fish liver ferritins. AB - Ferritin from the liver of fresh, salt and brackish water fishes was purified by thermal denaturation of liver homogenate followed by ammonium sulphate fractionation and Sephacryl S-300 gel filtration. Yield and iron content of purified fish ferritins were 0.016-0.026 mg/g of wet tissue and 4-14%, respectively. The iron content of ferritins from marine and brackish species was higher than from fresh water species. The phosphate/iron ratio ranged from 0.5 to 1.8 and was higher than mammalian ferritins. The fish ferritins have 5-6% neutral carbohydrate. Native gel electrophoresis and molecular weight analysis revealed the presence of a monomeric ferritin. SDS-gel electrophoresis and immunoblotting showed a single protein band of 21 kDa suggesting the presence of similar sized subunits in the native structure of fish ferritins. Isoelectric focusing revealed microheterogeneity with five to seven bands of pI values between 4.1 and 7.0. Variations in the amino acid composition were observed. Proline and arginine were not detected in murrel and salmon species, respectively. High proline and low tyrosine contents were recorded for perch ferritin. Immunological studies by non competitive indirect ELISA revealed varying degrees of cross-reactivity. Mammalian ferritins exhibited a moderate cross-reactivity with anti-fish ferritin. On the contrary, very low or no cross-reactivity was observed between fish ferritin and anti-mammalian ferritin. Ferritins from bony fishes such as murrel and rohu exhibited a high degree of cross-reactivity with anti-shark ferritin. However, a moderate cross-reactivity was observed between shark and anti-murrel ferritin. Ferritin from marine bony fishes, salmon and mackerel and perch (brackish) showed a low to very low cross-reactivity with both the antisera. PMID- 10481258 TI - Morphine coupling to invertebrate immunocyte nitric oxide release is dependent on intracellular calcium transients. AB - Morphine significantly stimulated invertebrate immunocyte intracellular calcium level increases in a concentration-dependent manner in cells preloaded with Fura 2/AM. Morphine's action was blocked by prior exposure of the cells to the opiate receptor antagonist naloxone. Various opioid peptides did not exhibit this ability, indicating a morphine-mu 3 mediated process. In comparing the sequence of events concerning morphine's action in stimulating both [Ca2+]i and NO production in these cells, we found that the first event precedes the second by 42 +/- 7 s. The opiate stimulation of [Ca2+]i- was attenuated in cells leached of calcium. strongly suggesting that intracellular calcium levels regulate cNOS activity in invertebrate immunocytes. PMID- 10481259 TI - Expression of porcine adipocyte transcripts: tissue distribution and differentiation in vitro and in vivo. AB - Transcription factor transcripts implicated in adipocyte differentiation (peroxisome proliferator-activated receptor gamma (PPAR gamma), retinoid x receptor alpha (RXR alpha), adipocyte determination and differentiation-dependent factor 1 (ADD1), and CCAAT/enhancer binding protein alpha (C/EBP alpha)) and adipocyte-characteristic protein transcripts (lipoprotein lipase (LPL) and adipocyte fatty acid binding protein (aP2)) were measured in pig tissues. Transcripts for PPAR gamma, ADD1, and aP2 were localized in porcine subcutaneous and perirenal adipose tissues; transcripts for C/EBP alpha and LPL were detected in other tissues, but the greatest concentrations were in the adipose tissues. In porcine stromal-vascular cells (S/V cells) differentiating in vitro, transcripts for PPAR gamma and aP2 increased gradually, transcripts for ADD1, and LPL increased early and transcripts for C/EBP alpha increased late. In pigs, adipose tissue transcripts for PPAR gamma, ADD1, and LPL were minimal at birth and increased to 28 days postpartum, transcripts for C/EBP alpha were low until 28 days and transcripts for aP2 were at high levels, regardless of age. Although transcript development was somewhat different in vitro and in vivo, the data suggest PPAR gamma (and ADD1 are involved in regulation of transcripts for LPL and that there may be more partially differentiated precursor cells in S/V cells at day 0 than in adipose tissue at birth. PMID- 10481260 TI - Structural and functional differences of two forms of GTP-binding protein, Gq, in the cephalopod retina. AB - The major GTP-binding protein (G-protein) in the rhabdomeric photoreceptor membranes of the squid (Watasenia scintillans) has been identified as a Gq-class G-protein. Anti-Gq alpha antibodies recognized a protein not only in the photoreceptor membranes but also in soluble fractions of the retina. The 42 kD protein in the soluble fractions (soluble Gq alpha) had the same molecular mass and the same reactivities to anti-Gq antibodies as those of membrane-bound Gq alpha. The G beta subunit was scarcely detected in the soluble fractions, being found mostly in the membrane fraction, indicating soluble Gq alpha exists in monomeric form. Soluble Gq alpha had no effect on the GTPase activity of the photoreceptor membranes, suggesting that it does not interact with photoactivated rhodopsin or G beta gamma. Soluble Gq alpha would be an inactive form of Gq alpha. In the retina of Octopus fangsiao, soluble Gq alpha was scarcely detected after dark adaptation, but increased during subsequent light exposure and decreased on returning to dark adaptation. These results with Octopus suggest that functional membrane-bound Gq alpha is converted to soluble Gq alpha on exposure to light. Transformation of membrane-bound Gq alpha into the soluble form by hydroxylamine suggests that the difference between membrane-bound and soluble Gq alpha is associated with the attachment of fatty acid(s). PMID- 10481261 TI - Simultaneous determination of trace benzodiazepines from drinks by using direct electrospray probe/mass spectrometry (DEP/MS). AB - This study presents a novel means of rapidly determining benzodiazepines in various drinks. Electrospray mass spectrometry with a direct probe design is used as the analytical instrument. Samples are treated only by a simple liquid-liquid extraction prior to direct electrospray probe/mass spectrometry (DEP/MS) analysis. The proposed method provides a relatively easy and efficient means of identifying the drugs in a very short time interval. On average, sample analysis is completed in less than five minutes including sample pretreatment. It is especially useful in forensic science since the technology can provide a rapid identification for unknown samples. Thus, the method proposed herein is highly appropriate for rapidly screening a large number of samples. PMID- 10481262 TI - Role of drugs and alcohol in impaired drivers and fatally injured drivers in the Strathclyde police region of Scotland, 1995-1998. AB - During the 4-year study period, 1995-1998, the Department of Forensic Medicine and Science, University of Glasgow received a total of 752 biological samples from drivers suspected of driving under the influence of drink and/or drugs in the Strathclyde region of Scotland. The majority of samples were blood and had been primarily obtained from males. Drugs were detected in 68 and 90% of blood and urine samples, respectively. Toxicological analyses revealed that cannabis was the most frequently encountered illegal drug which was detected in 39% of all drug positive blood samples. Benzodiazepines were detected in the majority of drug positive samples with 82% containing at least one member of this group. Polydrug use was prevalent, with the average number of drugs detected per sample increasing from 2.0 in 1995 to 3.1 in 1998. For comparison, the results of toxicological analyses from 151 fatally injured drivers are described. Although the majority of samples tested negative for the presence of drugs and alcohol, drugs were found to be present in 19% and alcohol was detected in 33%. As the majority of drugs had been prescribed or administered post-accident, this study shows that alcohol was the main causative factor conducive to fatal road traffic accidents. PMID- 10481263 TI - Sudden natural death 'at the wheel': a retrospective study over a 15-year time period (1982-1996). AB - In order to investigate the incidence and implications of sudden natural death at the wheel, a retrospective study was carried out over a 15-year time period ranging from 1982 until 1996. During this time period 147 drivers of motor vehicles were found out of 34,554 cases examined at the Institute of Legal Medicine, Ludwig-Maximilians University, Munich, Germany. There were 13 females and 134 males and the mean age was 56.8 years (range 20-86 years). The main cause of death was ischemic heart disease which was found in 113 cases. There were mainly minor injuries to the driver, or to other passengers, or to other road users and only minor property damage. Our study confirms previous investigations that sudden natural death at the wheel is a rare event in proportion to unnatural death at the wheel and is not a substantial threat to other road users. Despite existing guidelines for granting a driving licence, medical screening to exclude high risk patients from driving cannot prevent the occurrence of sudden natural death at the wheel. PMID- 10481264 TI - A review of the methodological aspects of aspartic acid racemization analysis for use in forensic science. AB - Accurate age determination of adult cadavers and human remains is a key requirement in forensic practice. The current morphological methods lack accuracy and precision, require specialist training and are costly. The use of aspartic acid racemization (AAR) in human dentine provides a simple, cost-effective solution and the method can achieve accuracies of +/- 3 years at best. Currently, there are differences in AAR methodology between laboratories which produce different results on the rate of racemization in teeth. These inconsistencies must be resolved if the technique is to be successfully applied to age determinations in forensic cases. This paper reviews the differences in protocol which have been used, discusses how each method will affect the results obtained from AAR analysis and gives recommendations for optimization of the methological protocol as a first step towards international standardization. PMID- 10481265 TI - Correlation between prescription of various dextropropoxyphene preparations and their involvement in fatal poisonings. AB - In Sweden, the frequency of fatal poisoning by dextropropoxyphene (DXP) ingestion is constantly high. There are seven preparations containing DXP on the Swedish market; in three of them DXP is the sole analgesic ingredient, while four of them are combinations of analgesics. In an attempt to assess the death rate attributable to each DXP preparation on the basis of toxicological analyses, altogether 834 cases of dextropropoxyphene-related death over a 5-year period (1992-1996) in Sweden have been reviewed. The ratio between number of fatal poisonings and prescription of defined daily dose/1000 inhabitants during a 12 month period (DDD) was determined. The highest ratio, 27, was attributed to unmixed preparations. The ratio for DXP + paracetamol-related deaths was 6.3, and for DXP + phenazone, 6.4, while the lowest ratio, 2, was found among the DXP + chlorzoxazone cases. The unmixed preparations, representing 26% of all DXP prescriptions during the study years, were implicated in 62% of the DXP fatalities, a considerable over-representation. Unmixed preparations, with their higher content of DXP, may be more attractive for many consumers because of their narcotic (euphoric) effects rather than for any analgetic superiority. Another possibility is that unmixed preparations may erroneously have been regarded as safer than when combined with paracetamol, as reports of poisoning with compounds containing DXP + paracetamol have been most frequently reported, probably due to their predominance on the market. PMID- 10481266 TI - Usefulness of conventional blood groups, DNA-minisatellites, and short tandem repeat polymorphisms in paternity testing: a comparison. AB - A total of 215 paternity cases were analysed after testing 24 marker systems. Despite technical advantages of polymerase chain reaction related polymorphisms (automatisation, employment of robots, lesser requirements concerning of quality and quantity of DNA) it could be shown that the exclusive employment of a parentage testing kit is compromised by an increased risk of erroneous conclusions. It is estimated that in about 3-4% of the cases ambiguous situations have to be expected which are caused by the occurrence of single or double exclusions. In these cases it is impossible to decide whether the exclusions indicate either true nonpaternity or a de novo mutation. The situation might become even more complicated if an involvement of a close relative of the alleged father cannot be ruled out. We cautiously advance the hypothesis that in parentage testing DNA minisatellite polymorphisms from an optimal set of tools. PMID- 10481267 TI - Epidermal growth factor pathway substrate 15, Eps15. AB - Eps15 was originally identified as a substrate for the kinase activity of the epidermal growth factor receptor (EGFR). Eps15 has a tripartite structure comprising a NH2-terminal portion, which contains three EH domains, a central putative coiled-coil region, and a COOH-terminal domain containing multiple copies of the amino acid triplet Aspartate-Proline-Phenylalanine. A pool of Eps15 is localized at clathrin coated pits where it interacts with the clathrin assembly complex AP-2 and a novel AP-2 binding protein, Epsin. Perturbation of Eps15 and Epsin function inhibits receptor-mediated endocytosis of EGF and transferrin, demonstrating that both proteins are components of the endocytic machinery. Since the family of EH-containing proteins is implicated in various aspects of intracellular sorting, biomolecular strategies aimed at interfering with these processes can now be envisioned. These strategies have potentially far reaching implications extending to the control of cell proliferation. In this regard, it is of note that Eps15 has the potential of transforming NIH-3T3 cells and that the eps15 gene is rearranged with the HRX/ALL/MLL gene in acute myelogeneous leukemias, thus implicating this protein in the subversion of cell proliferation in neoplasia. PMID- 10481268 TI - Laminin-11. AB - Laminins are a family of glycoproteins which are ubiquitous components of basement membranes and play key structural and functional roles. Eleven isoforms have been identified to date; each is an alpha beta gamma heterotrimer assembled from a repertoire of five alpha, three beta and two gamma chains. Studies of laminin-11 (alpha 5 beta 2 gamma 1) illustrate the unique expression patterns and distinct functions that can be displayed by laminin isoforms. Laminin-11 is found in the glomerular basement membrane in kidney, in the neuromuscular synaptic cleft in skeletal muscle and in other tissues such as placenta and lung. Mice lacking laminin-11 exhibit defective glomerular filtration and developmental defects in neuromuscular synapse formation, with Schwann cells invading the synaptic cleft. Consistent with these observations, both motoneurons and Schwann cells distinguish laminin-11 from other isoforms in vitro. These results suggest that laminin-11 is a structural component of the basement membrane which influences cell behavior in physiologically relevant ways. A greater understanding of laminin-11 assembly and basement membrane incorporation could provide a logical basis for therapy in merosin-deficient congenital muscular dystrophy. PMID- 10481269 TI - Tissue transglutaminase: an enzyme with a split personality. AB - Tissue transglutaminase (tTG) belongs to the family of transglutaminase enzymes that catalyze the posttranslational modification of proteins via Ca(2+)-dependent cross-linking reactions. The catalytic action of tTG results in the formation of an isopeptide bond that is of great physiological significance since it is highly resistant to proteolysis and denaturants. Although tTG-mediated cross-linking reactions have been implicated to play a role in diverse biological processes, the precise physiological function of the enzyme remains unclear. Recent data, however, suggest that the protein polymers resulting from tTG-catalyzed reactions may play a role in commitment of cells to undergo apoptosis. On the same token, tTG-mediated formation of insoluble protein aggregates may underlie the markers of numerous pathological conditions, such as the senile plaques in Alzheimer's disease and the Lewy bodies in Parkinson's disease. In addition to catalyzing Ca(2+)-dependent cross-linking reactions, tTG can also bind and hydrolyze guanosine triphosphate and adenosine triphosphate. By virtue of this ability, tTG has been identified as a novel G-protein that interacts and activates phospholipase C following stimulation of the alpha-adrenergic receptor. The ability of tTG to mediate signal transduction may contribute to its involvement in the regulation of cell cycle progression. The following review summarizes the important features of this multifunctional enzyme that have emerged as a result of recent work from different laboratories. PMID- 10481270 TI - Inhibition of 6-methyladenine formation decreases the translation efficiency of dihydrofolate reductase transcripts. AB - Cycloleucine was used to inhibit the formation of internal N6-methyladenosine residues in the messenger ribonucleic acid transcripts from cultured methotrexate resistant mouse sarcoma cells. Cells cultured in cycloleucine produced transcripts deficient in N6-methyladenosine residues and the 2'-O-methylated nucleosides of the cap structure; however, the formation of the 7-methylguanine nucleoside of the cap was not effected. Cytoplasmic polyadenylated transcripts were isolated from cells which had been pretreated with media containing cycloleucine and translated in an in vitro translation assay. The levels of translated dihydrofolate reductase were then analyzed by polyacrylamide gel electrophoresis. The amount of dihydrofolate reductase protein produced from the transcripts of the cycloleucine treated cells was 20% less than untreated transcripts. Ribonuclease protection assays demonstrated little difference in the cytoplasmic levels of dihydrofolate reductase transcripts between treated and untreated cells suggesting that the decrease in translation efficiency was not caused solely by an alteration in the processing or cytoplasmic transport of the transcripts. Translation of in vitro transcribed transcripts showed the presence of 2'-O-methylated nucleosides in the cap structure had a negative effect on translation efficiency, demonstrating that the results observed from cycloleucine treatment could not be due to the inhibition of 2'-O-methylation in the cap. These experiments therefore suggest that an inhibition of N6-methyladenosine residues in dihydrofolate reductase transcripts significantly alters their rate of translation. PMID- 10481271 TI - A comprehensive analysis of heat shock protein synthesis in human peripheral lymphocytes: the effect of penicillin/streptomycin. AB - A reliable experimental procedure is described for the simultaneous characterisation of a comprehensive range of heat shock proteins (hsps) in human peripheral lymphocytes. In this system, a mild heat shock from 37 to 42 degrees C for 1 h induced the synthesis of hsps 105, 90, 70, 60, 57, 47, 40, 27 and 16. Densitometric analyses of 35[S]-methionine labelled protein gels indicated that levels of these hsps peaked at 3 to 4 h, following post-heat shock recovery at 37 degrees C. The presence of penicillin and streptomycin in the cell culture medium, appeared to have little effect on the kinetics of hsp synthesis. The present method can be used for relatively small blood samples and its relative ease of application and reproducibility make it appropriate for screening the expression of hsps in human lymphocytes from a range of individuals. PMID- 10481272 TI - Molecular cloning, expression, localisation and functional characterisation of a rabbit SULT1C2 sulfotransferase. AB - The importance of sulfotransferases in xenobiotic metabolism is gaining recognition. The gastrointestinal (GI) tract is a major portal of entry for many xenobiotics, yet little is known about the contribution of sulfotransferases to detoxication or bioactivation metabolism in these tissues. To this end, isolation and characterisation of sulfotransferases expressed in the stomach of rabbits was undertaken. A unique sulfotransferase cDNA (GenBank Accession No. AF026304) was isolated from a rabbit stomach cDNA library. This cDNA was 1439 base pairs (bp) long and has an open reading frame of 888 bp. On expression of the cDNA in both COS cells and E. coli, a protein molecular weight of 34 kDa was detected on SDS PAGE. Immunoblotting using an antibody raised in goats against the bacterially expressed protein detected expression of the protein in GI tract tissues. The 34 kDa immunoreactive band was detected in rabbit GI tract tissues (stomach, duodenum, jejunum, ileum, colon, caecum and rectum), liver and kidneys, but not in the lungs (n = 3). The human ortholog (GenBank Accession No AF026303) of the rabbit enzyme was cloned from a human stomach cDNA library. These two enzymes share 84% amino acid sequence identity and have been termed 1C2 sulfotransferases. When functional and kinetic characterisation of the recombinant rabbit and human proteins was carried out using 16 known ST substrates, detectable sulfonation activity was observed only with p-nitrophenol (with Km values of 2.2 mM and 13.3 mM, respectively). In conclusion, we have identified a rabbit GI tract sulfotransferase belonging to a newly defined sulfotransferase subfamily. PMID- 10481273 TI - NMR analysis of the structure of synaptobrevin and of its interaction with syntaxin. AB - Synaptobrevin is a synaptic vesicle protein that has an essential role in exocytosis and forms the SNARE complex with syntaxin and SNAP-25. We have analyzed the structure of isolated synaptobrevin and its binary interaction with syntaxin using NMR spectroscopy. Our results demonstrate that isolated synaptobrevin in largely unfolded in solution. The entire SNARE motif of synaptobrevin is capable of interacting with the isolated C-terminal SNARE motif of syntaxin but only a few residues bind to the full-length cytoplasmic region of syntaxin. This result suggests an interaction between the N- and C-terminal regions of syntaxin that competes with core complex assembly. PMID- 10481274 TI - Hybrid-hybrid matrix structural refinement of a DNA three-way junction from 3D NOESY-NOESY. AB - Homonuclear 3D NOESY-NOESY has shown great promise for the structural refinement of large biomolecules. A computationally efficient hybrid-hybrid relaxation matrix refinement methodology, using 3D NOESY-NOESY data, was used to refine the structure of a DNA three-way junction having two unpaired bases at the branch point of the junction. The NMR data and the relaxation matrix refinement confirm that the DNA three-way junction exists in a folded conformation with two of the helical stems stacked upon each other. The third unstacked stem extends away from the junction, forming an acute angle (approximately 60 degrees) with the stacked stems. The two unpaired bases are stacked upon each other and are exposed to the solvent. Helical parameters for the bases in all three strands show slight deviations from typical values expected for right-handed B-form DNA. Inter nucleotide imino-imino NOEs between the bases at the branch point of the junction show that the junction region is well defined. The helical stems show mobility (+/- 20 degrees) indicating dynamic processes around the junction region. The unstacked helical stem adjacent to the unpaired bases shows greater mobility compared to the other two stems. The results from this study indicate that the 3D hybrid-hybrid matrix MORASS refinement methodology, by combining the spectral dispersion of 3D NOESY-NOESY and the computational efficiency of 2D refinement programs, provides an accurate and robust means for structure determination of large biomolecules. Our results also indicate that the 3D MORASS method gives higher quality structures compared to the 2D complete relaxation matrix refinement method. PMID- 10481275 TI - Heterologous expression of a deuterated membrane-integrated receptor and partial deuteration in methylotrophic yeasts. AB - Methylotrophic yeast has previously been shown to be an excellent system for the cost-effective production of perdeuterated biomass and for the heterologous expression of membrane receptors. A protocol for the expression of 85% deuterated, functional human mu-opiate receptor was established. For partially deuterated biomass, deuteration level and distribution were determined for fatty acids, amino acids and carbohydrates. It was shown that prior to biosynthesis of lipids and amino acids (and of carbohydrates, to a lower extent), exchange occurs between water and methanol hydrogen atoms, so that 80%-90% randomly deuterated biomass and over-expressed proteins may be obtained using only deuterated water. PMID- 10481276 TI - An NMR solution study of the mega-oligosaccharide, rhamnogalacturonan II. AB - Rhamnogalacturonan II (RG-II) is a structurally complex pectic mega oligosaccharide that is released enzymatically from the primary cell wall of higher plants. It contains roughly 30 monosaccharide units (MW approximately 5 kDa) including very unusual residues such as Kdo, Dha, aceric acid and apiose. Previous studies have demonstrated that these monomers are arranged into four structurally well-defined oligosaccharide side chains (A-D), linked to a homogalacturonan mainchain, but the specific attachment sites of these branches on the pectic backbone have not yet been elucidated. In the present work, fairly complete assignments of the 750 MHz 1H NMR spectra and partial assignments of the 13C NMR spectra of the sodium-borohydride-reduced RG-II monomer were obtained for a 5 mM sample isolated from red wine. On the whole, these data corroborate the primary structures of the sidechains previously established by methylation analysis, partial hydrolysis and FAB-MS spectrometry but some heterogeneity has been demonstrated (partial substitution at B5, B6, and A5). The preferred orientations of the majority of the sidechain glycosidic linkages in the RG-II monomer have been determined from the sequential nOe data and the solution structure is generally in good agreement with the stable conformers previously obtained by molecular modeling (MM3) of the disaccharide and sidechain oligosaccharide building blocks. All of a two-residue, a three-residue, and a four-residue segment of the backbone have been tentatively identified from long range interactions between sidechain protons as well as in the mainchain. Taking into account the length of the 9-mer galacturonan mainchain described in prior work, these building blocks constitute almost the complete structure of RG-II (Scheme 2). PMID- 10481277 TI - 1H and 15N chemical shift assignments for domain 4 of the common beta-chain of the IL-3, IL-5 and GM-CSF receptors. PMID- 10481278 TI - 1H, 13C, and 15N NMR backbone assignments of 37 kDa surface antigen OspC from Borrelia burgdorferi. PMID- 10481279 TI - Backbone 1H, 13C, and 15N resonance assignments of Streptomyces subtilisin inhibitor. PMID- 10481280 TI - 1H, 13C and 15N NMR assignments for a carbon monoxide generating metalloenzyme from Klebsiella pneumoniae. PMID- 10481281 TI - 1H, 13C and 15N resonance assignments and secondary structure of the N-terminal domain of human tissue inhibitor of metalloproteinases-1. PMID- 10481282 TI - 1H, 15N and 13C resonance assignments for the bromodomain of the histone acetyltransferase P/CAF. PMID- 10481283 TI - Assignment of the backbone resonances of oxidized Fe-superoxide dismutase, a 42 kDa paramagnet-containing enzyme. PMID- 10481284 TI - Sequential assignment and secondary structure analysis of the NADP(H)-binding domain of Escherichia coli transhydrogenase. PMID- 10481286 TI - Pathogenicity of Hansenula anomala in model of immunocompromised mice. AB - Systemic infections caused by opportunistic fungi have shown an increased frequency in the past 10 years, particularly in immunocompromised patients. Hansenula anomala is an ascosporogenous yeast of the Ascomycetes class found in the skin, throat, and digestive tract transient normal flora. This study was conducted to compare the pathogenicity of H. anomala and Candida albicans in a model of immunocompromised mice. Thirty-eight Swiss mice were divided into two groups as follows: 30 animals received an intraperitoneal (i.p.) injection of cyclophosphamide (200 mg/kg) four days before the induction of infection with H. anomala (1 x 10(6) yeasts/mL), and 8 animals received 100 mg/kg of cyclophosphamide at 3-day intervals during 3 weeks before inoculation of 1 x 10(7) yeasts/mL. All animals were treated with amoxicillin/clavulanic acid (40 mg/kg) four days before induction of infection. A group of mice inoculated with C. albicans (ATCC 64548) served as control. Tissue samples from the lung, spleen, liver, and kidney for histological and mycologic studies were obtained at necropsy. In each animal, the number of viable yeasts per gram of kidney was determined. The organs most frequently infected by H. anomala were the kidneys and the liver (20%), and the lung (10%). However, in conditions of sustained immunosuppression, H. anomala was found in 65.5% of the organs examined. It is concluded that in an experimental model of immunocompromised mice, the pathogenicity of H. anomala was low. PMID- 10481285 TI - Pathogenicities and GP43kDa gene of three Paracoccidioides brasiliensis isolates originated from a nine-banded armadillo (Dasypus novemcinctus). AB - We studied three different isolates of Paracoccidioides brasiliensis obtained from the mesenteric lymph node (D3LY1), the spleen (D3S1) and the liver (D3LIV1) of the same armadillo (Dasypus novemcinctus). Pulmonal inflammatory area was evaluated by intravenous inoculation of 10(6) yeast cells of each isolates in young, male, ddY mice. Moreover, the partial sequence of GP43kDa gene of P. brasiliensis was analyzed. The lung inflammatory area was greater in animals inoculated with isolate D3S1. The partial sequence of GP43kDa gene indicated that isolate D3S1 is different from isolates D3LY1 and D3LIV1. This study suggested that the same armadillo might be susceptible to multiple P. brasiliensis isolates simultaneously. PMID- 10481287 TI - Isolation of Hanseniaspora uvarum (Kloeckera apiculata) in humans. AB - Isolation of Hanseniaspora uvarum, a yeast of the ascomycetes group, whose anamorph corresponds to Kloeckera apiculata, obtained from stool and two ungual specimens from three patients, is reported. This yeast has been found in soil, water, various fruits, bivalve molluscs, crabs, prawns and fruit flies; in Spain, it has been described in the fermentation processes of some wines. In our region, it has also been found in the intestine of mackerel (Scomber scombrus). Its finding in humans constitutes a clinical rarity. PMID- 10481288 TI - Survey of termite-inhabited soil and mosquito breeding sites in Lucknow, India for potential mycopathogens of Anopheles stephensi. AB - During a short survey of soil and mosquito breeding sites in Lucknow, India for potential mycopathogen from a period of August-October 1996, 11 species of fungi in 5 genera were isolated using live mosquito larvae as host, Aspergillus flavus, A. fumigatus and Fusarium semitectum were the most frequently isolated species. Other fungi recorded were A. niger, A. ochraceus, A. terreus, A. versicolor, Geotrichum candidum, Penicillium verrucosum, Paecilomyces sp. and Fusarium sp. (Liseola/Elegans complex). Insect cell walls are known to contain chitin, so fungal isolates were tested for their chitinase activity on semi synthetic medium containing colloidal chitin. High chitinolytic activities were observed with A. flavus and A. ochraceus. Chitinase producers can be considered as potential pathogens. However, the higher incidence of F. semitectum could not be explained by inability to utilize chitin. PMID- 10481289 TI - Relationship between Fusarium graminearum and Alternaria alternata contamination and deoxynivalenol occurrence on Argentinian durum wheat. AB - A mycological survey was carried out on durum wheat (Triticum durum) samples from the main production area of Argentina. The isolation frequency and relative density of species of dematiaceous fungi, and genus Fusarium were calculated. Alternaria alternata and Fusarium graminearum were the predominant fungal species. An analysis of deoxynivalenol (DON) natural contamination was also performed on a limited number of samples (60). DON contamination levels in positive samples ranged from 26 to 6400 micrograms/kg. The non-parametric techniques applied showed that there is a positive relationship between DON contamination and F. graminearum relative densities and a negative relationship between DON contamination and A. alternata relative densities. PMID- 10481291 TI - Synthesis of sterigmatocystin derivatives and their biotransformation to aflatoxins by a blocked mutant of Aspergillus parasiticus. AB - Seven alkyl and aryl homologues of O-methylsterigmatocystin (OMST) were synthesised and fed in separate experiments to a mutant of Aspergillus parasiticus capable of converting sterigmatocystin (ST) to aflatoxin B1 (AFB1). Their conversion to AFB1 was followed over a time period and it was found that O propylsterigmatocystin (OPRST) was converted to AFB1 more rapidly than O ethylsterigmatocystin (OEST) or OMST or ST itself. The aryl derivative O benzoylsterigmatocystin (OBzST) was converted at the slowest rate. These results show that alkyl and aryl homologues of OMST may be converted to AFB1, suggesting that the methylation of ST is not an absolute requirement for its conversion to AFB1. It seems likely that whatever enzyme(s) are involved in this process exhibit relative specificity. As to whether alkylation of ST is an obligatory step in AFB1 biosynthesis is neither supported nor disproved as the fungal cells used are presumably capable of methylating ST. The fact that the propyl derivative showed fastest conversion is not necessarily significant as this may be due to faster diffusion of the least polar of the derivatives through the cell membrane. PMID- 10481293 TI - Heavy metal distribution in different fish species from the Mauritania coast. AB - Cadmium, copper, zinc and mercury concentrations were determined in pelagic and benthic fishes from the Mauritania coast. The pelagic fishes consisted of the round sardinella Sardinella aurita (Clupeidae), the chub mackerel Scomber japonicus (Scombridae) and the Atlantic horse mackerel Trachurus trachurus (Carangidae). Four benthic species were considered, three Serranidae: the painted comber Serranus scriba, the golden grouper Epinephelus costae, and the Niger hind Cephalopholis nigri and one Mullidae: the West African goatfish Pseudupeneus prayensis. Cadmium, copper, zinc and mercury concentrations are low in the edible muscles of the pelagic species. In the benthic fishes, metals could be determined also in the gills and liver. Cadmium and copper were present in relatively low levels in the muscle (< or = 0.06 microgram Cd g-1 dry wt., < or = 1.6 micrograms Cu g-1 dry wt.) and gills (< or = 0.23 microgram Cd g-1, < or = 3.1 micrograms Cu g-1) and higher levels are encountered in the livers (< or = 51 micrograms Cd g 1, < or = 49.1 micrograms Cu g-1). Zinc concentrations in the muscle are low (< or = 20 micrograms Zn g-1 dry wt.), the concentrations in the gills (< or = 120 micrograms Zn g-1) approach those of the livers for C. nigri and P. prayensis or are higher in the case of S. scriba and E. costae. Mercury concentrations in the gills and muscle of the pelagic species are very low, higher quantities are found in the livers of the benthic species. The metal concentrations in all the fishes analysed, which are restricted to approximately 40 specimens, are low except those found for cadmium in the livers of the benthic fishes. PMID- 10481292 TI - Monitoring regional fluoride pollution in the Saxonian Ore mountains (Germany) using the biomarker dental fluorosis in roe deer (Capreolus capreolus L.). AB - The spatial variation of fluoride contamination in the Saxonion part of the Ore mountains (Federal state of Saxony, Germany) was assessed by analysing the prevalence and severity of dental fluorosis in wild male roe deer aged 2 years or more (n = 794) from this region. The study area is exposed to increased atmospheric deposition of fluorides, originating from emission sources in the North-Bohemian brown coal belt. The severity of dental fluorosis in the permanent premolars and molars of one hemimandible per animal was assessed based on a scoring system and a dental lesion index of fluorosis (DLI) assigned to each specimen. On the basis of the mean DLI calculated for the roe deer originating from a certain municipal district, the respective deer sample was assigned to one of seven fluorosis categories, and a map showing the distribution of these categories was produced. Highest fluorosis categories were recorded for roe deer samples from the central and eastern parts of the study area that directly bordered on, or were located near to, the Czech-German border. A close spatial relationship between the main fluoride emission sources in North-Bohemia and the regions with the highest prevalence and severity of dental fluorosis in roe deer was discernible. The observed regional variation of fluoride pollution is in accordance with the results of other studies directly measuring fluoride deposition into the study area or fluoride levels in vegetation. A main advantage of the method used in this study over other ways of recording fluoride contamination of wildlife habitats is that it uses an established and continuously operated system of specimen collection, thereby enabling efficient monitoring with high spatial and temporal resolution at very low cost. Moreover, a clearly recognisable and relevant biological effect is measured which occurs in a dose-dependent manner over a range of environmentally relevant concentrations. PMID- 10481290 TI - Mycotoxin production and cytotoxicity of Fusarium strains isolated from Norwegian cereals. AB - Thirty-four isolates of the eight most common Fusarium species isolated from Norwegian cereals; F. avenaceum, F. culmorum, F. equiseti, F. graminearum, F. poae, F. sporotrichioides, F. torulosum and F. tricinctum were studied for their cytotoxicity and ability to produce mycotoxins. The strains were cultivated on rice, and analysed for trichothecenes (all species), zearalenone (all species), fusarochromanone (F. equiseti), wortmannin (F. torulosum), moniliformin and enniatins (F. avenaceum, F. tricinctum and F. torulosum). The cytotoxicity of the extracts were examined with an (in vitro) MTT-cell culture assay. All F. graminearum and five of seven F. culmorum isolates belonged to chemotype IA, producing deoxynivalenol and 3-acetyl-deoxynivalenol, while the two other F. culmorum strains were nivalenol producers (chemotype II). The F. equiseti isolates and one of the F. poae isolates produced both type A and B trichothecenes, and relatively large quantities of fusarochromanone were detected in the F. equiseti cultures. All Fusarium species studied showed significant cytotoxicity, but with a large variation between species, and also within each species. F. sporotrichioides and F. equiseti showed the highest average cytotoxicity. PMID- 10481294 TI - The characteristic study of TSP, PM2.5-10 and PM2.5 in the rural site of central Taiwan. AB - The total suspended particle (TSP), PM2.5-10 (aerodynamic diameter less than 10 microns) and PM2.5 concentration (aerodynamic diameter less than 2.5 microns) concentrations were sampled by PS-1 and Universal sampler on the roof (25 m) of the Medical and Engineering Building in the campus of Hungkuang Institute of Technology (HKIT) which is located at a height of 500 m on Da Du Mountain. The results indicated that average TSP, PM2.5-10 and PM2.5 concentrations are 0.42, 0.34 and 0.019 mg/m3 in the day time, respectively and are 0.32, 0.26 and 0.017 mg/m3 in the night time, respectively. The ratios of PM2.5-10/TSP were from 76% to 85% and from 50% to 91% for day and night period, respectively. It indicated that the major composition in the total suspended particles was PM2.5-10 in the rural site. The relationship between TSP and PM2.5-10 is TSP = 1.16PM2.5-10 + 0.027 and TSP = 1.01 PM2.5-10 + 0.058 in the day and night time, respectively. The correlation coefficient (R2) is 0.98 and 0.97 for day and night period, respectively. The relationship between PM2.5-10 and PM2.5 is PM2.5 = 0.0005PM2.5 10 + 0.019 and PM2.5 = 0.037PM2.5-10 + 0.0076 in the day and night period, respectively. The correlation coefficient (R2) is 3E-5 and 0.67 for day and night period, respectively. The relationships between TSP, PM2.5-10, PM2.5 particle concentrations and wind speed (R2) in the day time are 0.71, 0.64, 0.43, respectively and are 0.83, 0.79, 0.57, respectively in the night time. The proposed reasons are that there are more activities caused by people (students) and natural living animals which absorbed some of the particles during the day time. Thus, the correlation coefficients for the night time are better than those of day time. The particle size distributions are both bimodel in the day and night time. The major peaks in the day time appear in the particle diameter between 0.031-0.056 micron and 3.16-5.62 microns in the day period and appear between 0.017-0.031 micron and 1.78-3.16 microns in the night period. The results indicate that the particle size distribution in the day time tends to be of larger particle size mode than the night time. PMID- 10481295 TI - Secondary exposure to dioxins through exposure to PCP and its derivatives. AB - This study discusses the potential exposure of occupational workers and general consumers in the European Union (EU) to polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) caused by exposure to pentachlorophenol (PCP) and its derivatives. A synthesis of reported exposure data relating exposure to PCP and its derivatives in an occupational setting has indicated that the PCDD/F intake for exposure in the 1990s averaged over a lifetime is likely to be in the order of 0.02-1 pg I-TEQ kg-1 bw day-1 with actual exposures more likely to occur at the lower end of the range, around the median of 0.16 pg I-TEQ kg-1 bw day-1. Workers who have experienced past exposure to PCP and its derivatives especially in the early to mid 1980s, will have been subjected to higher exposures to PCDD/Fs due to the generally higher concentrations of PCDD/Fs in PCP products at that time. Exposure to PCP and its derivatives via the food chain is judged to be the most significant intake route of PCDD/Fs into consumers. The ingestion of milk and dairy products obtained from cows grazed on pasture dressed with sewage sludge has the potential to raise the average daily intake of PCDD/Fs via the diet by about 40% if all foods consumed derived from sludge amended soil. To the extent quantifiable, exposure to PCP in an occupational setting contributed approximately 16% as a median to the overall background exposure to PCDD/Fs, while for consumers this contribution ranged from 2 to 60% depending on the exposure to sludge amended foods. PMID- 10481296 TI - Development of germ cell transplants: morphometric and ultrastructural studies. AB - Mouse-to-mouse transplants were studied at 10 min, 9 h, 24 h, 1 week, 1 month, 2 months, and 3 months post-transplantation. Data from a previous light microscope study were confirmed and extended using morphometric and ultrastructural techniques. As soon as 10 min after introduction of the germ cells from one mouse into the tubule lumen of a recipient mouse they developed relationships with small Sertoli cell processes. The extent of this surface-to-surface relationship increased in animals sacrificed up to 1 week post-transplantation. Most transplanted germ cells retained the characteristics of the donor germ cells after they had been isolated and pelleted. Nearly all transplanted cells eventually underwent phagocytosis by the recipient Sertoli cells. The presence of small apparent clones of germ cells after 1 week of transplantation indicated that some germ cells may divide and survive for short periods within the epithelium. No discernible qualitative subcellular changes in the host Sertoli cell accompanying the development of transplant spermatogenesis were noted. Macrophages were present in the region of the boundary tissue between myoid cells and appeared to increase in number in the peritubular tissue of transplanted testes. Images suggest that they migrated into the tubule to gain entrance to the lumen and there take on the form of activated macrophages. Some macrophages phagocytose sperm at 2 months and 3 months post-transplantation. A testis weight increase previously demonstrate to occur at 24 h post-introduction of germ cells was found to be due to an increase in the volume of the tubular lumen. The increase of lumen size at 24 h was not related to the volume of the injected material. It is suggested that the presence of injected cells, likely germ cells, in the tubule lumen stimulated increased secretion by the Sertoli cell. PMID- 10481297 TI - Chicken thrombocytes in culture: lymphocyte-conditioned medium delays apoptosis. AB - Chicken thrombocytes are nucleated cells, analogs to mammalian platelets. These cells are involved in hemostasis, phagocytosis and secretion of specific products. Most of the properties of avian thrombocytes have been established in experiments that employed recently isolated blood cells. Attempts to cultivate these cells for a long period of time under optimal culture conditions for peripheral blood cells were unsuccessful; thrombocytes died after 24 h of cultivation unlike macrophages cocultured with them. Here we investigate the reasons and type of thrombocyte death in culture. Thrombocytes were separated from peripheral blood of roosters and cultured for 48 h. The influence of different culture conditions on thrombocyte viability was studied. Cells were cultured as adherent cell monolayers or under agitation (preventing adherence), in the presence or lack of lymphocytes or their soluble factors, and various concentrations of fetal bovine serum. After 24 h in standard culture thrombocytes displayed cytoplasm and chromatin condensation, DNA cleaved into oligonucleosomal fragments and unaltered mitochondria. These results strongly suggest that thrombocytes suffer an apoptotic cell death in culture. Apoptosis could be delayed by culturing thrombocytes in the presence of lymphocytes or their soluble factors. PMID- 10481299 TI - The length of the cycle of seminiferous epithelium in goats (Capra hircus). AB - This is the first report in literature showing the length of the seminiferous epithelium cycle in goats. In the present study, the duration of spermatogenesis was estimated using intratesticular injections of tritiated thymidine. Animals were castrated at 4 h, 7 days, and 11 days after injections. The duration of each spermatogenic cycle in goats is 10.6 +/- 0.5 days (SEM). Considering that the total duration of spermatogenesis takes about 4.5 cycles of seminiferous epithelium, spermatogenesis was estimated to last 47.7 days. The approximate primary spermatocytes life span is 14.1 days, while spermiogenesis in goats lasts 14.9 days. Staging in goats was based on the tubular morphology, where 8 stages of the cycle are yielded for all species. The relative stage frequencies in goats, based on 400 seminiferous tubule cross sections for each animal were as follows: stage 1: 15.8 +/- 1.0%; stage 2: 12.8 +/- 0.5%; stage 3: 20.5 +/- 0.9%; stage 4: 10.7 +/- 0.7%; stage 5: 11.6 +/- 0.6%; stage 6: 9.3 +/- 1.1%; stage 7: 7.6 +/- 0.4%; stage 8: 11.7 +/- 0.6%. The pre-meiotic, meiotic and post-meiotic phases' relative frequencies were 49.1%, 10.7% and 40.2%, respectively. The duration of spermatogenesis in goats is very similar to that found in rams. PMID- 10481298 TI - Morphological and biochemical analyses of the salivary glands of the malaria vector, Anopheles darlingi. AB - Adult Anopheles darlingi salivary glands are paired organs located on either side of the esophagus. The male glands consist of a single small lobe. The female gland is composed of two lateral lobes, with distinct proximal and distal portions, and a medial lobe. The lobes are acinar structures, organized as a unicellular epithelium that surrounds a salivary canal. The general cellular architecture is similar among the lobes, with secretory material appearing as large masses that push the cellular structures to the periphery of the organ. Cells of the proximal-lateral lobes show asynchronous cycles of secretory activity and contain secretory masses with finely filamentous aspect. In the distal-lateral lobes, cells display synchronous cycles of activity, and have a dense secretory product with mottled pattern. Cells of the medial lobe have secretory masses uniformly stained and highly electrondense. Biochemical analysis of the adult female salivary glands revealed apyrase, alpha-glucosidase and lysozyme activities. Alpha-glucosidase and lysozyme activities are detected mostly in the proximal lobes while apyrase is mainly accumulated in the distal lobes. This differential distribution of the analyzed enzymes reflects a specialization of different regions for sugar and blood feeding. Thus, the morphological differences observed in the lobes correlate with functional ones. PMID- 10481300 TI - Depletion of immune effector cells induces myocardial damage in the acute experimental Trypanosoma cruzi infection: ultrastructural study in rats. AB - The contribution of radiosensitive cells and macrophages to myocardial immunopathology has been studied in rats inoculated with Trypanosoma cruzi, Y strain. Immunodepression was induced by gamma irradiation and depletion of radioresistant macrophages was achieved by silica, a selective cytotoxic agent for macrophages. Irradiated or silica treated rats and age-matched controls were sacrificed at day 12 of infection so as to study the heart by light and electron microscopy. In the infected controls, damaged cardiomyocytes were directly related to tissue parasitism; inflammatory cells, predominantly lymphocytes and macrophages, were present. The drastic depletion of radiosensitive cells (lymphocytes and granulocytes), as well as the depletion of macrophages by silica, induced cardiomyocytes damage during the acute infection, exacerbating the lesions seen in the infected controls. In the irradiated-infected and silica treated-infected animals, degenerating cardiomyocytes, parasitized or not, were frequently observed, displaying evident signs of cytoplasmic and nuclear damage. Some signs of cardiomyocyte damage (irregular distribution of glycogen particles and myofibrils with shrinkage and aggregation of Z bands) were present only in silica treated-infected animals. The findings suggest that immune effector cells may not play a major role in the cardiomyocyte damage induced by acute. Chagas disease, arguing against the autoimmune etiology of Chagasic cardiomyopathy. PMID- 10481301 TI - Distribution and space-time relationship of proteoglycans in the extracellular matrix of the migratory pathway of primordial germ cells in mouse embryos. AB - In this paper we present an in situ ultrastructural cytochemical study on the distribution and spatial-temporal expression of proteoglycans (PGs) in the extracellular matrix of the migratory pathway of mouse primordial germ cells (PGCs) during the different phases of migration, by the use of the cationic dye ruthenium hexammine trichloride (RHT). Embryos of 9, 10, 11 and 12 days of development were used. The treatment with RHT revealed PGs as electron dense layers, granules, and filaments. Whereas granules prevailed in the extracellular spaces of the migratory route during the whole migratory process, the amount of filamentous structures increased during the migration phase of PGCs. At the end of the migratory process the surface of the PGCs lost its reaction by RHT. There were differences in the size of the granules of PGs at the initial migratory period (9-day-old embryos) as compared with the other days of gestation. There was a strong reaction for PGs in the extracellular spaces, expressed as a meshwork of granules interconnected by filaments, as well as reaction on the basement membranes during the peak of the PGCs migration in 10-day-old embryos. These results support the hypothesis that these molecules may have an important role in the migration of PGCs, although the precise mechanism involved in this process is not yet clear. PMID- 10481302 TI - The epiphyseal cartilage and growth of long bones in Rana catesbeiana. AB - The structure of the epiphyseal cartilage of the bullfrog Rana catesbeiana and its role in the growth of long bones were examined. The epiphyseal cartilage was inserted into the end of a tubular bone shaft, defining three regions: articular cartilage, lateral articular cartilage and growth cartilage. Joining the lateral cartilage to the bone was a fibrous layer of periosteum, rich in blood vessels. Osteoblasts with alkaline phosphatase activity were found on the surface of the periosteal bone, which presented a fibrous non-mineralised tip. The growth cartilage was inside the bone. The proliferative chondrocytes presented perpendicular separation of daughter cells and there was no columnar arrangement of the cells. Furthermore, chondrocyte hypertrophy was not associated with either calcification or endochondral ossification, in apparent contrast to the avian and mammalian models. Finally, there was no reinforcement system capable of directing cell volume increase into longitudinal growth. Since bone extension depends on the intramembranous ossification of the periosteum, the growth cartilage is inside and not at the end of the bone and the cells in the growth cartilage show no columnar arrangement and separate in a direction perpendicular to the long bone axis, we conclude that the growth cartilage mainly contributes to the radial expansion of the bone. PMID- 10481303 TI - A comparative study of lymph node mast cell populations in five marsupial species. AB - In order to determine whether different subpopulations of mast cells exist, mast cells of mandibular and axillary lymph nodes from five species (Didelphis aurita, Metachirus nudicaudatus, Philander opossum, Marmosops incanus and Gracilinanus agilis) of South American marsupials were studied. Our results showed that mast cells present in the connective tissue of the capsule and septa (CTMC) were similar to those described for eutherian mammals. However, a population of mast cells that was present in the lymphatic sinuses and bathed by the lymph, plus in direct contact with granulocytes, lymphocytes, macrophages, and reticular cells, were morphologically and histochemically different from the CTMC. In the five species studied, these cellular types, called lymphatic-sinus mast cells (LSMC), had a lower concentration of intragranular heparin and, in four of the five species, the cytoplasmic granules appeared to be larger than those in CTMC. Although LSMC have been rarely described in eutherian mammals, it was verified, in this study, that LSMC are nevertheless present in lymphatic sinuses of the five metatherian species studied. These observations suggest that the presence of LSMC may be a characteristic of the marsupials and important in the immune response and adaptive success of the Didelphidae. PMID- 10481304 TI - Seminiferous epithelium cycle and its duration in capybaras (Hydrochoerus hydrochaeris). AB - Although capybara is the largest rodent in the world and largely distributed in Central and South America, there is no report in the literature concerning the cycle of seminiferous epithelium in this species. In the present study, the length of spermatogenic cycle was estimated using intratesticular injections of tritiated thymidine. Animals were sacrificed at 1 h, 8 days, and 17 days after injections. The duration of one spermatogenic cycle in capybaras is 11.9 +/- 0.1 days (SEM). Spermatogenesis was estimated to last 53.6 days, when considering that the total duration of spermatogenesis takes about 4.5 cycles of seminiferous epithelium. The approximate life span of primary spermatocytes is 19.1 days, while spermiogenesis lasts 16.7 days. Staging in capybaras was based on the spermatid nuclei shape and location of spermatids, named tubular morphology method, which consists of 8 stages in all species. The relative stage frequencies in capybaras, based on the analysis of approximately 200 cross sections of seminiferous tubule for each of the ten animals were as follow: stage 1: 14.0 +/- 1.5%; stage 2: 15.1 +/- 1.0%; stage 3: 15.7 +/- 1.1%; stage 4: 14.6 +/- 1.1%; stage 5: 8.7 +/- 0.7%; stage 6: 7.0 +/- 0.7%; stage 7: 9.4 +/- 0.9%; stage 8: 15.5 +/- 1.0%. The pre-meiotic, meiotic and post-meiotic phases relative frequencies were 44.8%, 14.6% and 40.6%, respectively. Compared to most rodents investigated so far, the duration of spermatogenesis in capybaras is relatively long. PMID- 10481305 TI - Paracrystalline structures in the epithelial principal cells of the epididymis of water buffalo (Bubalus bubalis). AB - The ultrastructure of many principal cells in the cauda epididymis of water buffaloes with ages varying between 4, 18, 24, 30 and 36 months, revealed, in many cells, the presence of long, curved paracrystalline structures that are quite large and frequently encountered in the cytoplasm, usually near the nucleus. Concomitantly or not, smaller rod-like, hexagonal or curled structures can be found in the nucleus. Both structures, cytoplasmic and intranuclear, are made up of a sheath of parallel filaments. These paracrystals may appear as thin, regularly spaced filaments that are associated with fine, evenly spaced subunits. Occasionally, the association of paracrystalline structures with membranes similar to the endoplasmic reticulum was observed, but no membranes were consistently found in close contact with the nuclear crystalloids. It is postulated that both structures are proteinaceous and may represent stored enzymes or substances present in the intraluminal fluid, which are absorbed and initially stored in numerous intraepithelial vacuoles of the corpus and cauda of the buffalo epididymis. PMID- 10481306 TI - Immunocytochemical mapping of the hemoglobin biosynthesis site in amphibian erythroid cells. AB - During the past 25 years, several studies have attempted to determine the site of integration of the heme and the four globin chains in vertebrate erythroid cells that is important in the formation of the hemoglobin molecule. Mitochondrion-like organelles or hemosomes were pointed out as responsible for this task. We performed several experiments to investigate this hypothesis. The intracellular distribution of hemoglobin in amphibian erythroid cells was detected by post embedding immuno-electron microscopy, using a polyclonal anti-human hemoglobin proteinA-gold complex. Hemoglobin mapping showed an intense labeling in the cell cytoplasm, but none in cytoplasmic structures such as endoplasmic reticulum, mitochondria, mitochondrion-like organelles, Golgi complex, ribosomes or ferruginous inclusions. The mitochondrial fraction obtained according to the protocol described for some authors, showed by ultrastructural examination that this fraction has a heterogeneous content, also composed by microvesicles rich in cytoplasmic hemoglobin, an artifact generated by mechanical action during cell fractionation. Thus, when this fraction is lysed and its content submitted to electrophoresis, hemoglobin bands would be found inevitably, causing false positive results, erroneously attributed to hemoglobin content of mitochondrion like organelles. Our data do not confirm the hypothesis that the final hemoglobin biosynthesis occurs inside mitochondrion-like organelles. They suggest that the hemoglobin molecule be assembled in the erythrocyte cytoplasm outside of mitochondria or hemosomes. PMID- 10481307 TI - Structural elements common to mitosis and apoptosis. AB - Both mitotic and apoptotic cells display hypercondensation of the chromatin and loss of the nuclear envelope (Lazebnik et al., 1993). Herein, we describe a third similarity between the two processes. We have observed, initially in apoptotic cells of the PC-12 lineage clusters of 40-60 (approximately 50) nm vesicles adjoined by a minor contingent of tubule vesicular elements of 100-200 nm which are indistinguishable from their vesicular counterparts in mitotic PC-12 cells. The clusters of approximately 50 nm vesicles were subsequently observed in all studied rat tissue cells in apoptosis (plasma cells and macrophages, secretory epithelial cells from pancreatic acini, ventral lobe of prostate and mammary gland). Clusters of approximately 50 nm vesicles comparable to those of the PC-12 cells were found in HeLa cells treated with human alfa TNF, in WEHI-3 cells exposed to VM 26 (a teneposide) (Sesso et al., 1997) and in HL-60 cells treated with thapsigargin. PC-12 and HeLa cells affixed to coverslips were double labelled and examined with the fluorescence microscope to reveal simultaneously the disposition of the chromatin with Hoechst stain and the distribution of the fluorescence of Golgi or of Golgi-associated proteins. A common pattern of fluorescence was observed in a minor proportion of apoptotic cells using three different antibodies used. The label frequently appeared as finely dispersed granules in the cytoplasm. In some apoptotic cells, relatively coarse granules were observed. This pattern of label distribution is compatible with the disposition of vesicular clusters we have encountered in apoptotic PC-12 cells sectioned serially or semi serially. In such sections of both mitotic and apoptotic PC-12 cells, we noticed that the conglomerates of 50 nm vesicles were frequently associated with cisternae of the rough ER. Vesicles of similar size were also noted pinching off from the extremities of Golgi cisternae reduced in size. These cisternae diminish in length and width when they are in the process of disassembling at the very beginning of mitosis and in apoptosis. PMID- 10481308 TI - Exposure to constant light during testis development increases daily sperm production in adult Wistar rats. AB - Testis histometry and daily sperm production (DSP) were evaluated in adult (160 day-old) Wistar rats exposed to constant light for the first 25 days after birth, and compared with control animals which were exposed to a 12 h-light-12 h-dark light regimen. Significantly greater (P < 0.05) numbers of Sertoli cell nucleoli and round spermatids per cross-section of seminiferous tubule were found in animals exposed to constant light. In addition, epididymis weight, DSP per testis and per gram of testis, as well as Leydig cell compartment volume, were significantly increased in treated animals. Although there was a clear trend toward an increased Sertoli cell population per testis in animals exposed to constant light, this difference was not statistically significant (P < 0.05). The number of round spermatids as expressed per Sertoli cell was the same in both groups. Surprisingly, the diameter and volume of round spermatid nucleus at stages I and VII of the cycle of seminiferous epithelium were significantly lower (P < 0.05) in treated animals. In conclusion, constant illumination during neonatal testis development increased sperm production and Leydig cell compartment volume in adult rats probably through a mechanism involving elevated follicle stimulating hormone and luteinizing hormone during the prepubertal period. To our knowledge, this is the first study showing that altering the light regimen can affect sperm production in non-seasonal breeders. PMID- 10481309 TI - Transgenic Arabidopsis leaf tissue expressing a modified oryzacystatin shows resistance to the field slug Deroceras reticulatum (Muller). AB - Transgenic Arabidopsis thaliana has been developed which expresses the oryzacystatin mutant OC-I delta 86, which is an inhibitor of the major proteinase present in the digestive gland of the slug, Deroceras reticulatum. When fed on leaf tissue from plants expressing this inhibitor the growth of juvenile slugs was significantly reduced by 31% compared with those feeding on control leaf tissue. Furthermore, while surviving slugs did not individually consume less when feeding on leaf tissue expressing OC-I delta 86, the total amount of leaf tissue eaten was 50% less, due to reduced survival of slugs. The synthetic cysteine proteinase inhibitors E-64 and leupeptin also significantly reduced slug weight gain (by at least 40%) and digestive gland cysteine proteinase activity when administered in an artificial diet, indicating that their antimetabolic effects are due to direct inhibition of gut proteolytic activity. These results suggest that transgenic crop plants expressing phytocystatins could be used to suppress the growth rates of slug populations in the field. PMID- 10481310 TI - Type II fish antifreeze protein accumulation in transgenic tobacco does not confer frost resistance. AB - Type II fish antifreeze protein (AFP) is active in both freezing point depression and the inhibition of ice recrystallization. This extensively disulfide-bonded 14 kDa protein was targeted for accumulation in its pro- and mature forms in the cytosol and apoplast of transgenic tobacco plants. Type II AFP gene constructs under control of a duplicate cauliflower mosaic virus 35S promoter, both with and without a native plant transit peptide sequence, were introduced into tobacco by Agrobacterium tumefaciens-mediated transformation. AFP did not accumulate in the cytosol of transgenic plants, but active AFP was present as 2% the total protein present in the apoplast. Plant-produced AFP was the same size as mature Type II AFP isolated from fish, and was comparable to wild-type AFP in thermal hysteresis activity and its effect on ice crystal morphology. Field trials conducted in late summer on R1 generation transgenic plants showed similar AFP accumulation in plants under field conditions at levels suitable for large-scale production: but no difference in frost resistance was observed between transgenic and wild-type plants during the onset of early fall frosts. PMID- 10481311 TI - Isolation and microinjection of somatic cell-derived mitochondria and germline heteroplasmy in transmitochondrial mice. AB - At present, there are no means for creation of relevant animal models of human mitochondrial DNA (mtDNA)-based diseases in a directed fashion. As an initial step towards this end, we have developed a microinjection technique for transfer of isolated, viable mitochondria between two mouse species. Previously, we reported detection, by nested PCR with species-specific primer sets, of Mus spretus mtDNA in Mus musculus domesticus blastocyts following zygote microinjection and culture. We now report the production of transmitochondrial founder mice, and germline transmission of the heteroplasmic state in a maternal lineage. Heteroplasmic mice produced by this technique will be useful in the study of mitochondrial dynamics and may hasten the creation of animal models of human mtDNA-based diseases. PMID- 10481312 TI - Over-expression of the murine polymeric immunoglobulin receptor gene in the mammary gland of transgenic mice. AB - The polymeric immunoglobulin receptor (pIgR), a transmembrane protein, transports dimeric IgA (dIgA) across the epithelial cells of the mucosal surfaces into the external secretions, for example milk from the mammary glands. The pIgR is consumed during the transcytosis of dIgA and is cleaved at the apical side of the epithelial cells, regardless of the binding to its ligand (dIgA), to form secretory component (SC). We hypothesize that the expression level of the endogenous murine pIgR gene in the epithelial cells is rate-limiting for the transport of dIgA across the epithelial cells into the secretions. We address this key issue by generating transgenic mice over-expressing the pIgR gene in their mammary glands in order to examine the effect on dIgA levels in the milk. Here we report on the generation of transgenic mice and analysis of the expression level of pIgR in their mammary glands. We cloned and characterized the murine pIgR gene and constructed an expression cassette bearing the pIgR gene under the control of the regulatory sequences of the bovine alpha s1-casein gene. Four transgenic lines were made, expressing the pIgR construct at RNA and protein level only in their mammary glands. The levels of the SC protein in the milk ranged from 0.1 to 2.7 mg/ml during mid-lactation. These levels are 10-270 times higher than wild-type SC levels (0.01 mg/ml). PMID- 10481313 TI - Transfer of chloramphenicol-resistant mitochondrial DNA into the chimeric mouse. AB - The mitochondrial DNA (mtDNA) chloramphenicol (CAP)-resistance (CAPR) mutation has been introduced into the tissues of adult mice via female embryonic stem (ES) cells. The endogenous CAP-sensitive (CAPS) mtDNAs were eliminated by treatment of the ES cells with the lipophilic dye Rhodamine-6-G (R-6-G). The ES cells were then fused to enucleated cell cytoplasts prepared from the CAPR mouse cell line 501-1. This procedure converted the ES cell mtDNA from 100% wild-type to 100% mutant. The CAPR ES cells were then injected into blastocysts and viable chimeric mice were isolated. Molecular testing for the CAPR mutant mtDNAs revealed that the percentage of mutant mtDNAs varied from zero to approximately 50% in the tissues analyzed. The highest percentage of mutant mtDNA was found in the kidney in three of the chimeric animals tested. These data suggest that, with improved efficiency, it may be possible to transmit exogenous mtDNA mutants through the mouse germ-line. PMID- 10481314 TI - Linearization and purification of BAC DNA for the development of transgenic mice. AB - Bacterial artificial chromosome (BAC) vectors are increasingly used for generation of transgenic mice due to the relatively large size and the stability of their inserts compared to YACs. We have compared methods for purification and linearization of BACs, and describe an optimised protocol for preparation of high quality linear BAC DNA based on lambda terminase digestion, electroelution of linearized DNA together with simple preliminary multiplex PCR screening to detect transgenic mice. Linearized BAC DNA purified this way was successfully used for the development of transgenic mice containing 2-4 copies of the transgene. PMID- 10481315 TI - [The range of congenital malformations associated with cleft lip and palate]. AB - The congenital malformations associated to cleft lip and cleft palate cover a wide range of pathologies with incidence and acuteness that vary according to the authors. In the 284 patients followed and/or referred to the Cleft Consultation of Dona Estefania Hospital, 78 had associated malformations (27.5%). In three cases there was consanguinity in parents. A familial incidence of cleft and/or other congenital malformations was found in 13 and 10 patients respectively. Thirty patients had malformative syndromes. In the remaining 48, 127 associated congenital malformations (M.C.A.) were identified, in accordance with the Smith criteria, 81 major and 46 minor. The most frequent associated malformations are those of the face (25.9%) and of the cardiovascular system (16.5%). The multiple anomalies (of various systems) are the most frequent (47.9%), followed by the isolated anomaly (29.1%) and the multiple of one system (22.9%). In what concerns the association of syndromes with the type of cleft, primary, secondary or total palate, those of the secondary were the most frequent, particularly that of the Pierre Robin sequence (S.P.R.)--19 in 36 secondary cleft palate. PMID- 10481316 TI - [The determination of susceptibility to measles, rubella, mumps and chickenpox in students of health-related areas]. AB - We present an epidemiological and serological study in 286 health care students. We found susceptibility for measles in 11.7% individuals (95% confidence interval (95% CI): 8.0-15%), for rubella: 6.7% (95% CI: 3.8-9.6%) for mumps: 12.7% (95% CI: 8.0-16.6%) and for varicella 8.5% (95% CI: 5.3-11.7%). Compared to a similar study, performed in 1992 in a population of health care workers, we found an increasing susceptibility to these diseases except for mumps, that had decreased. Among those who received one dose of measles vaccine we found 12.1% non immune. We found an high level of immunity (97.1%) for those who received rubella vaccination. We could not draw any conclusions for mumps because only seven had been vaccinated. PMID- 10481317 TI - [The repercussions of mental disease in the family. A study of the family members of psychotic patients]. AB - Caregiving and related concepts such as family or caregiver burden have been widely discussed in recent literature. A descriptive, cross-sectional study was conducted in a sample (n = 80) of primary caregivers to severe mental patients (diagnosed schizophrenia, chronic psychosis or affective disorder). OBJECTIVES: To describe the caregivers' and corresponding patients' sample, as well as the nature of existing relationships. Levels of caregiver burden and psychological distress were evaluated and a preliminary analysis conducted on objective and subjective burden correlation. METHODS: Involvement Evaluation Questionnaire (IEQ), General Health Questionnaire (GHQ) and a semi-structured interview designed for this purpose. RESULTS: The extent of burden was found significant in this sample, being unevenly distributed in its components (e.g. tension, worrying or financial burden). Forty-one percent of caregivers experienced minor psychiatric morbidity. Measures of objective and subjective burden were moderately correlated. The same applied to magnitude of associations between each of them and distress parameters. Higher levels of burden were found in groups of caregivers to more severely ill or disabled patients, while diagnosis was not important for this purpose. CONCLUSIONS: Most relatives remain highly involved. These results suggest the need to implement specific interventions addressing psychotic patients as well as their families. PMID- 10481318 TI - [The diversity of Staphylococcus aureus strains isolated in a Lisbon hospital over a 4-year period]. AB - Over a 4-year period, 2020 Staphylococcus aureus strains isolated in Santa Maria Hospital were studied, 26.3% of which were methicillin-resistant (MRSA). The main specimens from which the strains were isolated included pus, blood and sputum/bronchial secretions. Isolation in blood cultures was the most common source among patients from medical units. Antimicrobial susceptibility studies showed that while in methicillin susceptible strains sensitivity to other antimicrobial agents (apart from penicillin resistance) was the rule, in MRSA strains there was resistance to most antibiotics. Only vancomycin was active against all strains. Phage typing showed that 75.5% of the strains were typable with phages at 100 x R.T.D. Among methicillin sensitive strains, a big diversity of phage patterns was observed, including phage groups I, II, III and V, as well as with phage association D11/95. The large majority of MRSA strains were lysed by group III phages, although several distinct patterns were observed. Within these strains, lysis by groups II and V phages was not observed. Plasmid profiling was the least discriminant issue in the characterization of these micro organisms because most of the strains harboured only one plasmid (or none). These results showed that a dominant MRSA strain did not exist in this hospital, but rather several distinct strains. The importance, as well as the difficulties in controlling the spread of MRSA strains in the present conditions of high prevalence, are highlighted. PMID- 10481319 TI - [The application of the endovascular prosthesis (stent)]. AB - We report two cases of symptomatic internal carotid stenosis treated by vasoplasty and stent placement in two male patients aged 57 and 64 years, one who had been suffering from transient ischemic events and the other had had a recent cerebral infarction with hemiparesis. The treatment sessions took place without complications. We discuss the choice of simultaneous vasoplasty and stent placement, some of the current possible indications for the method and the importance of multidisciplinary planning for the patient's evaluation and treatment. PMID- 10481320 TI - [Pelvic presentation]. AB - Labour is very important in breech presentation and it is associated to significant morbidity and mortality enhancement in recent years. The authors made a retrospective study of breech presentation and labour with this kind of fetal presentation, in Santa Maria Hospital in 1993 and analysed the following data: maternal age, parity, gestational age, the mode of delivery, the fetal weight, the maternal and fetal morbidity and mortality. The authors also reviewed the Literature on the incidence, the most frequent complications and the management in labour to compare with the results of the study. The authors found that the incidence of breech presentation in this hospital is slightly higher (4.7%) than the universal data. The remaining results had no significant difference. PMID- 10481321 TI - [Trigeminal neuralgia]. AB - The lesion of the sensorial nerves of the face is expressed by painful phenomena called neuralgias of the brainstem or its branches. These may be idiopathic or essential and secondary or symptomatic and their clinical evidence and course are different from the former. Essential neuralgia (E.N.), or tic douloureux, is the most frequent facial pain neuralgia. The aim of this paper is to review this subject with evidence of secondary trigeminal neuralgia from oral causes (dental, traumatic, post herpes zoster, etc). The author establishes the differential diagnoses with other etiologies of facial pain and mentions the diagnostic criteria for trigeminal neuralgia (T.N.), makes the difference between essential neuralgia (E.N.) and symptomatic neuralgia (S.N.); describes briefly the different kinds of the medical and surgical therapy, its management and complications. The author finishes by asserting that all E.N. must complementary means of diagnosis (Base skull, X rays, C.T. scan, M.R.I.). Peripheral surgery has more relapses and less morbidity than neuro-surgical techniques. Under these circumstances latter must considered by the patient's, or his relatives, opinion. PMID- 10481322 TI - [Parvovirus B19 infection]. AB - In 1975, during blood screening for hepatitis B, Cossart et al. discovered the human parvovirus B19 (B19). It is a small, single strand DNA virus of the Parvoviridae family. This virus is widespread with 40-80% of adults showing evidence of infection. It is found in the respiratory secretions of viraemic patients and direct contact has been suggested as the most likely mode of transmission. Parenteral transmission is common during treatment with clotting factor concentrates, but rarely occurs during transfusion with single donor products. Although B19 usually causes a self-limited illness, complications of infection can be severe and at times life threatening. In pregnant women, infection can lead to spontaneous abortions and hydrops fetalis and, in patients with haemolytic anaemias or in immunocompromised individuals, can induce aplastic crisis and chronic anaemias. The diagnosis can be made by indirect (testing for B19 antibodies) or direct methods (detecting B19 viremia). There are no vaccines or specific therapy currently available. Contact isolation is recommended for hospitalized patients. PMID- 10481323 TI - [The treatment of cytomegalovirus retinitis in human immunodeficiency virus infection]. AB - Most of the patients with immunodeficiency virus infection (75%-85%) are coinfected with cytomegalovirus. It is estimated that cytomegalovirus disease develops in 45% of them, mainly those presenting low CD4 cell counts (< 100/mm3). Retinitis is the most common manifestation of reactivated cytomegalovirus disease (85%), followed by other extra ocular sites (central nervous system, lungs, gastrointestinal system, adrenal glands, etc.). The particularly high incidence of cytomegalovirus retinitis, the difficulties concerning its therapeutical approach and the relatively unsatisfactory results, justify this review, according to the new treatment options presented in recent literature. PMID- 10481325 TI - [The hemolytic-uremic syndrome]. AB - The microangiopathic thrombotic syndromes--thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS)--are characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal dysfunction, fever and central nervous system abnormalities. Today they are considered as two extremes of a continuous spectrum named TTP--HUS. The syndrome is an uncommon disease with a high mortality rate, despite treatment. The authors describe a case of hemolytic uremic syndrome in a young adult patient. Initially the clinical course and the first biopsy suggested a favourable prognosis, but the early recurrence with severe hypertension was followed by a fatal outcome 6 months later. Concerning this clinical case, the authors present a review of the most recent aspects of the pathogenesis and treatment of this syndrome. PMID- 10481326 TI - [Hemoperitoneum as the form of presentation of hepatocellular carcinoma. The contribution of computed tomography to the diagnosis]. AB - Haemoperitoneum following rupture of hepatocellular carcinoma (HCC) is unusual. Two patients with secondary massive intraperitoneal haemorrhage due to spontaneous rupture of HCC diagnosed by Computed Tomography (TC) are discussed. The most important clinical and imagiological features of this pathology are presented. PMID- 10481324 TI - [Fractures of the mandibular condyle. Therapeutic controversies]. AB - The condylar mandibular fractures are important because its incidence, possible complications and controversial treatment. The treatment of condylar fractures has generated more controversy and discussion than any other in the field of maxillofacial trauma. The main goal of treatment is restoration of function and not anatomic restoration of parts. Despite several clinical and anatomical studies still lack consensus regarding the best method of treatment. This review article focus on the controversy that surrounds treatment of the condylar fractures, trying to supply consensus about questions like: Should condylar mandibular fractures be managed via a closed or open technique? What is the best surgical approach? Surgical timing? What is the degree and duration of mandibular immobilization? Is or not necessary to treat the ATM disc? PMID- 10481327 TI - [Type-II dyserythropoietic anemia. A partial form of the glycoprotein degradation syndrome?]. AB - The authors present a case of a boy, aged 8 years and 11 months, yellow race, with dyserythropoietic anemia type II, diagnosed at two months of age. Screening for partial form of carbohydrate deficient glycoprotein syndrome was normal. This result did not confirm the publication by Fukuda in 1990. PMID- 10481328 TI - [The embolization of complicated renal angiomyolipoma]. AB - A case of complicated renal angiomyolipoma with tumoral bleeding in a 44-year-old woman is presented. The contribution of angiography to the diagnosis helped clarify the degree of vascularity, and the contribution of embolization treatment (safe and well tolerated) is discussed. PMID- 10481329 TI - [A lymphoepithelioma-like type of spinous-cell carcinoma of the cervix uteri]. AB - Lymphoepithelioma-like carcinomas of the cervix uteri are very rare. They are poorly differentiated squamous cell carcinomas with intense stromal lymphocytic infiltration. These histologic features are similar to nasopharyngeal lymphoepithelioma and may have a better prognosis than other tumors of the cervix. A lymphoepithelioma-like lesion of the cervix uteri is described in a 33 year-old Caucasian woman who had an episode of vaginal bleeding. A review of the literature about these types of tumor is also presented. PMID- 10481330 TI - [Chlamydia pneumoniae and arteriosclerosis: an innocent onlooker?]. PMID- 10481331 TI - [Septic polyarthritis: treatment complications and results]. AB - OBJECTIVE: The rare polyarticular clinical presentation of peripheric septic arthritis has worst prognosis than monoarticular one. We studied 10 patients with polyarticular septic arthritis treated during the last 8 years, to evaluate functional result and complications after surgical treatment. MATERIAL AND METHODS: We studied 5 men and 5 women, with an average age of 46.5 years (range 18-75 years), they presented septic arthritis in two or more joints. We discuss aetiology, clinical presentation and treatment. RESULTS: Treatment was surgical debridement and parenteral antibiotic therapy. Functional result was poor in 54% of the joints treated, 50% of patients presented septic shock and three died. PMID- 10481332 TI - [An endemic goiter study of a population in the Pyrenees (Cerdanya-Girona)]. AB - INTRODUCTION: To evaluate the iodine deficiency, the scholar population is the most usually studied. However, the survey in general population offers information about the most affected groups. Our work was made in the frame of a health survey of Cerdanya (12,200 inhabitants), and five years after salt iodination. AIMS: Aims.--1) Prevalence evaluation of endemic goiter and urinary iodine excretion in Cerdanya population (> 6 y.). 2) Measurement of iodine concentration in drinking water. MATERIAL AND METHODS: 492 individuals were randomly recruited (46.5% men and 53.5% women). A questionnaire and estimation of thyroid size (Perez) were performed. Urinary iodine excretion was determined (Benotti) in 25% of the subjects. Measurement of iodine concentration in drinking water were performed. RESULTS: Goiter prevalence (> or = Ia) was 28.7% (IC 95%: 24.8-32.6). The prevalence was higher in women (35.3% vs 21%/p < 0001). Goiter prevalence > Ia was 7.9% (IC 95%: 5.6-10.2) and for nodular goiter 3.4% (IC 95%: 1.9-4.9). In our population the goiter prevalence = Ia, decreased with age, but no significant changes age-related for goiter > or = Ib was observed. In women > or = 45 years and previous pregnancy, the prevalence was higher (35.6% vs 17.6%. p < 0.05). Median urinary iodine was 165 micrograms/l. In 81% of the population the fish consumption was higher than once a week. Mean iodine concentration in drinking water was 2.04 mg/l (2.1 SD). CONCLUSIONS: After promote iodinated-food consumption, good results with a decrease in goiter prevalence and a increase in urinary iodine excretion were observed. Pregnancy increases the risk of goiter development. Although young people was more affected, the good level of urinary iodine excretion was the best indicator to expect progressive goiter disappearance. The low iodine concentrations in Cerdanya water drinking, explains the iodine deficiency in this area. PMID- 10481333 TI - [Community-acquired bacteremic pneumonia in the elderly]. AB - OBJECTIVE: To assess the risk factors associated with the development of bacteremic community-acquired pneumonia in the elderly (BCPE) and its clinical and prognostic aspects. METHODS: We studied 36 BCPE and 72 patients with community-acquired pneumonia and negative blood cultures as control group. RESULTS: The presence of the following risk factors was more common in cases than in control subjects: alcoholism, malnutrition, diabetes and chronic renal insufficiency. The multivariate analysis demonstrated that the malnutrition (OR:3.3; CI95%:1.4-7.8) and diabetes (OR:2.5; CI95%:1.1-6.7) were independent risk factors significantly related to the development of BCPE. The presence of chills, tachycardia, tachypnea, decreased level of consciousness and hypotension blood pressure were more common in case subjects. The microbial agents implicated more frequently were S. pneumoniae and gram-negative bacilli. The bilateral involvement and two or more affected lobes on chest radiograph were more common in the cases of BCPE. The mortality rate was higher in case subjects (22.2% vs. 2.8%; p = 0.001). CONCLUSIONS: We stress the relevance of the bacteremia as prognostic factor in the community-acquired pneumonia in the elderly. PMID- 10481334 TI - [Video thoracoscopy and video-assisted thoracic surgery. An analysis of 303 consecutive cases]. AB - OBJECTIVES: To present our experience in video thoracoscopy and video-assisted thoracic surgery from November of 1991 until November of 1997. PATIENTS AND METHODS: 303 endoscopic thoracic surgery procedures (210 males and 93 females with a mean age of 42.2 years) with the following indications: 151 spontaneous pneumothorax, 51 pulmonary biopsies, 50 pulmonary nodules, 15 mediastinal tumors, 13 pleuroparietal tumors, 8 pericardial windows and 15 thoracic sympathectomies. RESULTS: Mean postoperative stay was of 5.5 days. The thoracotomy conversion was necessary in 32 cases (10.9%). The morbimortality has been: one parietal recidive of a tumor in the way of one of the trocars and one death of a patient to who was accomplished a pulmonary biopsy by diffuse pneumopathy. CONCLUSIONS: The video thoracoscopy and thoracic surgery are effective and sure technical options for diagnosis and treatment of various thoracic affections. PMID- 10481335 TI - [Malignant pleural mesothelioma with an unusual radiologic manifestation]. AB - The malignant pleural mesothelioma is a rare neoplastic consequence observed in people with previous exposition to asbestos essentially. The malignant mesothelioma like a term, not only reports to primary malignant extended tumors that are derived of pleural mesothelioma but also, pericardial and peritoneal (about 20%). The exposition of asbestos stands for a sequential cellular reaction with oncogenic potential and with a typical majority clinical presentation. We described the case a patient complaint of malignant pleural mesothelioma with unusual radiology presentation with the result that unilateral calcified pleural plaques with pleural thickening and pleural effusion absence. Definitive diagnostic was achieved by thoracotomy. PMID- 10481336 TI - [Rare presentations of thyroid carcinoma]. AB - The hyperthyroidism is associated to different clinical situations with a relative frequency on the daily practice. Anyway, there have been discovered and published some clinical cases of thyroid cancer with insufficient symptoms, associated with a typical clinical hyperthyroidism and that have been related with Graves Basedow Disease. We expose here two different kinds of non typical thyroid cancer as a contribution to medical literature. In these cases, thanks to a careful exploration, further tests and, above all, fine needle aspiration puncture, hidden cancer was discovered. Cancer was not expected because of clinical symptoms. The definitive treatment was marked by fine needle aspiration puncture. PMID- 10481337 TI - [Respiratory distress syndrome due to hyperleukocytic leukemias]. AB - Hyperleukocytic leukemias are a small proportion of leukemias that have white blood cell count > 100 x 10(9)/l, most of them are leukemic blast cells. These leukemias have a grave prognosis because they can develop a leukostasis syndrome which describes: the acute onset of pulmonary failure and, often, neurologic deficits and disseminated intravascular coagulation (DIC). The leukostasis is produced by the mechanical obstruction of vascular bed by blast cells, which can be induced by the spontaneous tumor lysis or as a side effect of cytotoxic drugs. So, hyperleukocytic leukemias require early and vigorous measures to decrease the white blood cell count, using leukapheresis and/or chemotherapy, before pulmonary failure exists. Then, it is possible to reverse the lesions. We report two cases of acute myeloblastic leukemia with a white blood count > 100 x 10(9)/l, that developed a respiratory distress syndrome and died. The postmortem examination has been done in one of the cases. PMID- 10481338 TI - [7 cases of Staphylococcus lugdunensis infection]. AB - Staphylococci are frequently isolated from human, animal and environmental samples. The genus Staphylococcus comprises 27 species, of which 23 are coagulase negative. The pathogenic role of the coagulase-negative species remains to be fully established. Staphylococcus lugdunensis is a recently described coagulase negative species. Its Latin name reflects the city (Lyon, France) where the bacteria was first described. The present study reports 7 cases of human infection caused by S. lugdunensis. Case 1: a 64 y/o diabetic woman in whom S. lugdunensis was isolated in the conjunctival exudate. Case 2: a 51 y/o male diagnosed of a bladder tumor, with S. lugdunensis detected in the purulent exudate of a transurethral resection. Case 3: an 81 y/o woman with cerebrovascular stroke, in whom S. lugdunensis was isolated from urine. Case 4: a 65 y/o male with bacterial endocarditis and S. lugdunensis isolation in blood culture. Case 5: a 79 y/o male with liver cirrhosis and S. lugdunensis in blood culture. Case 6: a 75 y/o female with cerebrovascular stroke, in whom S. lugdunensis was isolated from urine. Case 7: a 77 y/o male diagnosed of femoral osteosarcoma and isolation of S. lugdunensis in the purulent exudate of the wound. The present communication broadens the range of clinical syndromes in which coagulase-negative S. lugdunensis appears as pathogen. PMID- 10481339 TI - [Citrobacter freundii endocarditis]. AB - Infection by Citrobacter appears in man only in certain circumstances, since it usually acts as contaminant or colonizer. Bacteraemia by this bacillus can affect immunodeficient people, elderly people or those patients who have undergone invasive hospital processes. Although incidence of bacteraemia is low (0.3-0.9%), the death rate is very high, about 48%. This bacillus is seldom the cause of endocarditis. That is why we describe a case of endocarditis by Citrobacter freundii, in an aged person with previous valvulopathy. PMID- 10481340 TI - [Sister Mary Joseph's nodule. Its clinical significance and management]. AB - The eponym Sister (Mary) Joseph's nodule is referred to the existence of umbilical metastasis. Historically it has been considered a sign of ominous vital prognosis and therapeutic abstention. The analysis of 407 cases published at the medical literature (MEDLINE 1966-1997) is against this idea, because recent series have showed survivals longer than a year after treatment. On the other hand 60% of umbilical nodules are benign. Nowadays this sign must be considered as an early indicator of inner neoplasms, mainly from the abdominal cavity (52% digestive origin, 28% gynecologic origin), and in 40% of cases they are the only expression of relapse of previously known neoplasms. When a umbilical nodule is found it is necessary to make an accurate histologic differential diagnosis between a primary (benign or neoplastic) and metastatic lesions. Fine needle aspiration has demonstrated to be an easy and reliable method for diagnosis. Once the metastatic origin of the nodule is confirmed treatment with curative intention must be tried. PMID- 10481341 TI - [Pulmonary complications related to cocaine consumption]. PMID- 10481342 TI - [Multiple pulmonary nodules as the form of presentation of tuberculosis in the adult]. PMID- 10481343 TI - [Pneumocystis carinii pneumonia in an immunocompetent patient after paraquat spraying]. PMID- 10481344 TI - [Pulmonary edema associated with upper airway obstruction]. PMID- 10481345 TI - [Laryngeal cancer and the leukemoid reaction]. PMID- 10481346 TI - [Streptococcus bovis sepsis, purulent meningitis and colonic diverticulosis]. PMID- 10481347 TI - [A new case of multifocal bone lymphoma]. PMID- 10481348 TI - [Can macrolide treatment delay the diagnosis of tuberculosis?]. PMID- 10481349 TI - [The carpal tunnel syndrome and hypothyroidism]. PMID- 10481350 TI - [Ginkgo biloba extract (EGb 761). State of knowledge in the dawn of the year 2000]. AB - EGb 761 is a standardized extract of dried leaves of Ginkgo biloba containing 24% ginkgo-flavonol glycosides, 6% terpene lactones such as ginkgolides A, B, C, J and bilobalide. Its broad spectrum of pharmacological activities allows it to be in adequacy to the numerous pathological requirements--hemodynamic, hemorheological, metabolic--which occur in cerebral, retinal, cochleovestibular, cardiac or peripheral ischemia. Moreover, EGb 761 has direct effects against necrosis and apoptosis of neurons and improves neural plasticity as evidenced in vestibular compensation. At the molecular and the cellular levels, some evidence obtained with animal models indicates that EGb 761 can interact as a free radical scavenger and a inhibitor of lipid peroxidation with all, or nearly all reactive oxygen species; maintains ATP content by a protection of mitochondrial respiration and preservation of oxidative phosphorylations; exerts arterial and venous vasoregulator effects involving the release of endothelial factors and the catecholaminergic system. Moreover, EGb 761 regulates ionic balance in damaged cells and exerts a specific and potent Platelet-activating factor antagonist activity. Numerous well-controlled clinical studies, realized in Europe and in USA, have revealed that EGb 761 is an effective therapy for a wide variety of disturbances of cerebral function, ranging from cerebral impairment of ischemic vascular origins (i.e. multi infarct dementia), early cognitive decline to mild to-moderate cases of the more severe types of senile dementias (including Alzheimer's disease) or mixed origins (i.e. psychoorganic origin). Improvement of signs and symptoms have been demonstrated for cognitive functions, particularly for memory loss, attention, alertness, vigilance, arousal and mental fluidity. Some clinical studies have showed that EGb 761 treatment may improve the capacity of geriatric patients to cope with the stressful demands of daily life. The explanation is a dual stress-alleviating action of EGb 761: its facilitates behavioral adaptation to stress and may decrease the excess of cortisol release to stress. Moreover, EGb 761 shows a specific neuroprotective effects to hippocampic cells. Regarding the visual system, experimental studies have shown that EGb 761 can inhibit or reduce the functional retinal impairments resulting from ischemia-reperfusion, photo-degeneration, diabetic or proliferative retinopathy. Clinical studies have revealed that EGb 761 may be useful in treating visual activity impairments and damages to the visual field associated with chronic cerebrovascular insufficiency, senile macular degeneration and diabete mellitus. Regarding the vestibular and auditory systems, experimental and clinical studies have shown the efficacy of EGb 761 in treating hypoacusis, tinnitus, vertigo, dizziness and other symptoms of vestibulocochlear disorders. At least, adequatly controlled studies in patients with peripheral arterial occlusive disease have provided good evidence for therapeutic efficacy in intermittent claudication. The future of EGb 761 is undoubtedly in the promise in slowing the progression of Alzheimer's disease. Indeed, two recent american clinical studies have shown the efficacy and safety of EGb 761 in patients with mild to severe Alzheimer's disease and multi-infarct dementia. In clinical terms, progression of symptoms was delayed by approximately 6 months. Actually new clinical studies are undertaken in USA and Europe. At the dawn of the third millenium (the Sixth for Ginkgo biloba) we propose a state of art about it. PMID- 10481351 TI - [The role of anti-leukotriene drugs in the management of chronic asthma]. PMID- 10481352 TI - [Atopic dermatitis on infant physical examination]. AB - The prevalence rate of atopic dermatitis was investigated in 4-month-old infants, who were examined by dermatologists at regular health checks at a health center. Among 739 infants examined, 150 were diagnosed as having atopic dermatitis, giving a prevalence rate of 20.3%. Most of the cases were mild, and no severe cases were observed. The prevalence rate in spring and autumn was significantly higher than that in summer and winter. The eruptions were seen mainly on the head, face, and trunk. A familial history of atopic dermatitis was positively correlated with infant atopic dermatitis in this series, whereas a familial history of bronchial asthma and allergic rhinitis showed no such correlation. It is important to make a correct diagnosis of mild atopic dermatitis by differentiation from other diseases by dermatologists, and it will be of interest to observe the condition of the infants when they attend for their next physical examination at the age of 18 months. PMID- 10481353 TI - [The increase and decrease of the total pollen counts of Sugi (Cryptomeria japonica) due to abnormal weather during pollen scattering season]. AB - We investigated the changing total number of airborne pollen grains of Sugi (Cryptomeria japonica) due to abnormal weather during pollen scattering season. The pollen season was divided into 7 periods by the discrepancies in forest flowering time for observations. The observation forests were located at an altitude of 150 m, 220 m, 350 m, and 500 m, and 800 m in the Rokko mountains. These were located at equal spaces, when altitude was taken in the logarithm. The distribution percentage of pollen counts from 1993 to 1998 showed an almost normal distribution level. This was demonstrated by the Log-Normal Probability paper, with a middle level altitude of a 350 m forest. We could estimate the relative rate of 5 different altitude forests against the corresponding amount of pollen as shown under, 0.09 at 150 m forest, 0.34 at 220 m forest, 0.31 at 350 m forest, 0.16 at 500 m forest, 0.10 at 800 m forest. The percentage of pollen counts at the 500 m forest in 1994 and at the 350 m forest in 1995 were decreased from 95% confidence interval. We regarded the decrease in the percentage of pollen counts during these two time, as a cause weak wind velocity and wrong wind direction in Tanba mountains. It was shown that the method for dividing pollen season in different observation forests in which altitude differs, will be useful for pollen forecasts and the pollen source countermeasure. PMID- 10481354 TI - [Tele-medicine system for high-risk asthmatic patients]. AB - We have developed a tele-medicine system to monitor the airway status at home for patients with poorly controlled asthma, whereby a nurse provides instructions to individuals via the telephone to help them manage exacerbation under the supervision of their physicians. We examined the effectiveness of this system with a randomized control study. Patients with high hospitalization risk were enrolled in the study by screening patients for those with multiple previous emergency room visits and randomly assigned to either the tele-medicine or control group. After six months of participation in the program, the number of emergency room visits decreased significantly and the activities of daily living were improved in the tele-medicine group. Most of the patients in the tele medicine group were able to continue measuring and transmitting peak expiratory flow (PEF) value successfully, and at six months had noticed an improvement in PEF. We therefore conclude that the system effectively contributes to the management of poorly controlled asthma. In addition, further consideration suggests that the reduction of emergency room visits may lead to reduction in hospitalization since we found a good correlation between number of emergency room visits and hospitalization from the studies published previously. PMID- 10481355 TI - [The diagnostic role of the methacholine inhalation challenge in adult patients with chronic persistent cough]. AB - Chronic persistent cough (CPC) is a common medical problem. We determined the value of the methacholine inhalation challenge (MIC) in the evaluation of CPC. We also sought other clinical factors that affect MIC. Patients were selected for the study if 1) CPC was the only presenting symptom, 2) a current roentgenogram did not contribute to the diagnosis, and 3) spirometry revealed a normal FEV1. We measured the minimum dose of methacholine (bronchial sensitivity, Dmin) by the "Asthograph" method. We determined the correlation between log Dmin and demographic and clinical variables, i.e. age, %FVC, %FEV1, FEV1%, %V50, %V25, eosinophil count in peripheral blood, and serum IgE level. The causes of CPC in 51 patients (20 men and 31 women, mean age 41 years) were cough-variant asthma in 29 patients, postinfectious persistent cough in 13 patients, atopic cough in 6 patients, and others in 3 patients. Log Dmin significantly but weakly correlated with %V25 (4 = 0.31, p = 0.02). The sensitivity and specificity of Dmin (< 10 units) in diagnosing cough-variant asthma was 93% and 87%, respectively. Demographic or clinical variables other than Dmin were not useful for the diagnosis of cough-variant asthma. We conclude that MIC is useful for the differential diagnosis of CPC while the usefulness is limited in some cases. PMID- 10481357 TI - [A relationship between birch pollen and meteorological factors during the past four years and pollen survey in Sapporo in 1997]. AB - It is well known that grass pollinosis has been the most representative pollinosis in Hokkaido prefecture, Japan, However, it is also well known that birch (Japanese white birch, Betula platyphylla var. japonica) pollinosis is gradually increasing, which is marked especially in Sapporo and its neighborhood. We analyzed a relationship between the birch pollen count and meteorological factors statistically. Moreover, the principal airborne pollen counts in Sapporo in 1997 were assessed and compared with the previous pollen data. As a results of this study, it was showed a close relationship between the annual amounts of birch pollen and meterological factors of the year before, especially in May and June might be important. Moreover, there is a strong negative correlation between the birch pollen count and humidity, which seems to affect bringing up flower buds. As for the principal airborne pollen counts, some kinds of tree pollen, including birch pollen, were observed from the end of March, followed by grass (Gramineae) pollen in June and mugwort (Artemisia) pollen from the end of August, respectively. The annual amounts of grass pollen showed a decreasing trend. PMID- 10481356 TI - [Role of Candida allergen in atopic dermatitis and efficacy of oral therapy with various antifungal agents]. AB - We studied whether fungal allergens play a role in exacerbating the clinical symptoms of atopic dermatitis (AD). We found that the percentage of the patients who showed CAP-RAST positive (2 < or = score) to Candida albicans (Ca) was significantly higher in the patients with severe symptoms and high serum IgE level than those with mild symptoms and lower IgE. This was also true for the patients with CAP-RAST positive to Pityrosporum ovale (Po). AD patients with their symptoms localized to head and neck showed significantly higher tendency to have positive CAP-RAST (2 < or = score) to Ca and Po when compared to those with their eruption distributing to the extremities. We then evaluated the efficacy of oral therapy with antifungal agents in 140 cases of refractory AD with positive CAP-RAST to Ca. Good or excellent response was obtained in 60% with fluconazole, 35% with itraconazole, 31% with amphotericin B, 28% with nystatin. The present finding that amphotericin B and nystatin, both of which are not absorbed through intestine, were effective for approximately a third of the patients indicates that Ca in the intestine plays an important role in triggering AD symptoms. Fluconazole was more effective than amphotericin B and nystatin, suggesting that fungal colonizing in other parts of the body but the digestive tract also play a role in the exacerbation of AD symptoms. PMID- 10481358 TI - [Significance of area under the flow volume curve--useful index of bronchial asthma]. AB - Some studies have reported that area under the flow volume curve (AUFVC) can be an index of pulmonary function. However, the significance of AUFVC remains to be clarified. We have clarified that AUFVC reflects the momentum of expired air. Size of flow volume curve (= AUFVC) has been commonly recognized to be changeable visually in patients with asthma. To clarify whether size of flow volume curve (= AUFVC) is a useful index of the course of bronchial asthma, we compared the rate of improvement forced expiratory volume in 1 second (FEV1.0), peak expiratory flow rate (PEFR) or forced vital capacity (FVC) to the rate of improvement in AUFVC after admission in 20 patients with bronchial asthma. The rate of improvement in AUFVC positively correlated with the rate of improvement in FEV1.0, PEFR or FVC. AUFVC demonstrated more marked improvement than other indices. Since AUFVC reflects the momentum of expired air, bronchial asthma's patients could understand visually that the momentum of expired air decreased due to airway stenosis by presenting flow volume curve. Therefore, visual size of flow volume curve helped patients with asthma to understand the condition of asthma. Size of flow volume curve (AUFVC) may be useful index of the course of bronchial asthma. PMID- 10481359 TI - Regrets. AB - A year or so ago I was invited by an Italian dentist to spend a day with him in his practice and to attend a two-day seminar, which he was presenting. The location, in the mountains near to Lake Como and the Swiss border, was enough to persuade me to accept. However I was to find out that the professional aspects of the trip were not only most invigorating but that they would also cause me to look back with concern to my own career. Had I done as well for my patients, my family and myself as I could have done? PMID- 10481360 TI - Long term risks of impacted molars. PMID- 10481361 TI - More to consider in nerve injury debate. PMID- 10481362 TI - Operator competence and nerve damage. PMID- 10481363 TI - Introduction to dental implants. PMID- 10481364 TI - A retrospective study of 50 treatments using an appliance to produce localised occlusal space by relative axial tooth movement. AB - This paper describes the use of 50 appliances which have been used to produce a localised occlusal space prior to the restoration of teeth. These were provided for 45 patients aged 19 to 69 years at the Eastman Dental Hospital London between 1981 and 1994. The concept is similar to that described by Dahl, the space being produced by the teeth in contact with the appliance being intruded and those out of contact over erupting. Many of the appliances however differed from those originally described by Dahl. PMID- 10481365 TI - Barriers to accessing dental care: patient factors. PMID- 10481366 TI - The consultant orthodontic service--1996 survey. AB - OBJECTIVE: To evaluate the working patterns and facilities of the consultant orthodontist service. DESIGN: A cross-sectional survey. SETTING: Consultant orthodontist departments in the UK. SUBJECTS AND METHODS: All consultant orthodontists in the UK were sent a questionnaire that gathered information on the individual consultant, the facilities available, the new patients referred and patients under current treatment. RESULTS: The consultant orthodontist service provided treatment to a high number of patients who were in definite need of orthodontic treatment. A marked reduction in the use of removable appliances suggests improving standards of care and provision of more complex treatment. The caseload was high and a fair proportion of patients were returned to their referring dentists with treatment plans. The consultant service has not completely evolved into a service that provides treatment at a super-specialist level alone. CONCLUSIONS: There has been little change in the consultant orthodontist service over the past ten years. Arguably, this is because of the wishes of the purchasers and the shortage of trained orthodontic manpower as a direct result of poor manpower planning and lack of funds for post-graduate training. PMID- 10481367 TI - Electro-chemically activated water in dental unit water lines. AB - OBJECTIVE: To investigate the effect of electro-chemically activated water on biofilm contamination in dental unit water lines. DESIGN: Thirteen dental units fitted with independent water systems and used for 12 years with distilled water were divided into two groups, A and B. At the start, one week later, and again four weeks later, the bacterial counts in water from all units were determined. Also specimens of tubing were taken from the units at the beginning and at the end of the study for SEM investigation. In Group A distilled water was replaced with electrochemically activated water (a Russian invention), and used continuously for the duration of the study. In group B, distilled water was used as before, until confirmed to be contaminated. For ethical reasons group B was treated, one week into the study with conventional disinfectants. SETTING: The project was carried out in a clinic of a department of periodontology of a faculty of dentistry during 1998. RESULTS: Both groups showed a marked reduction in bacterial counts. Under SEM Group A showed a total elimination of the biofilm and Group B a partial removal. CONCLUSIONS: Distilled water was ineffective in controlling bacterial counts and biofilm. Electrochemically activated water was effective for this purpose. PMID- 10481368 TI - Variations in planning fixed bridgework--a group of dentists at a case-based postgraduate course. AB - Variations in treatment planning and prescription have been described in relation to routine restorative dentistry. This study examined dentists' decisions regarding treatment planning for fixed bridgework. Fifty five dentists who attended a Continuing Education course on fixed bridgework were given standard information about a patient in the form of study casts, photographs of radiographs and a clinical history. They were asked to design a bridge where a previous one had failed and to complete a proforma which was returned to the course organisers in advance of the event. The response rate was 65%. The data showed wide variation in identification of features of diagnostic importance. Seventy percent of respondents chose to use again as abutments teeth which were extensively damaged and had failed to retain the previous bridge: while only 30% noted features of the occlusion which if left unchanged would have precluded a successful bridge from being made. A further feature was that nearly 70% chose to use multiple abutments to support the bridge. The implications of these results were discussed with respect to current concepts of bridge design. PMID- 10481369 TI - [Behavior of piglets during castration with and without local anesthesia]. AB - According to the German welfare act of 1998 (Art.5(3)1.), surgical castration of up to 4 wk old pigs does not require anaesthesia. However, the application of a local anaesthesia is discussed in the context of pain related responses to this procedure. In these experiments, effects of different methods of application on the behaviour of piglets during castration are evaluated as a contribution to the welfare assessment of this procedure. A total of 36 piglets between 10 and 14 days of age were castrated with and without a local anaesthesia. Local anaesthesia with Lidocaine 2% was applied in two ways: a) via intratesticular (i.t.) injection and b) intratesticular injection combined with subcutaneous (s.c.) infiltration of the anaesthetic into the tissue around the spermatic cord. The assessment of pain was performed on the basis of vocalisations and resistance movements of the animals during the procedure. Local anaesthesia resulted in a reduction of the relative proportion of resistance movements from the entire period of fixation (castration with i.t. anaesthesia 12.73%, with i.t. and s.c. anaesthesia 14.50% or without a local anaesthesia 18.96%; P < 0.05). In particular, local anaesthesia resulted in a reduction of resistance movements during the cutting of the spermatic cord. The response of piglets to additional skin stimulation of the scrotum was not different between the three treatments. PMID- 10481370 TI - [Prevalence and importance of endoparasites in calves raised in Swiss cow-calf farms]. AB - A longitudinal study was performed to establish estimates of prevalence of selected endoparasites in Swiss cow-calf operations and to investigate the importance of an infection with endoparasites on diarrhea and weight gain until weaning. Three hundred and eighty-six calves raised in 67 larger cow-calf herds were included in the study. Faecal samples were collected during the first three months of life and at weaning. Parasitological analyses were performed according to standard procedures including specifically the detection of Eimeria spp., Cryptosporidium parvum, Strongyloides papillosus and Trichostrongylida. The effects of an infection with endoparasites on weight gain were analyzed with a linear model accounting for effects of farm, breed, sex, calving month and weight at birth. The average prevalences of endoparasites within the first three months of life were: E. bovis 36%, E. zuernii, 19%, C. parvum 16.8%, S. papillosus 22.3% and Trichostrogylida 1.5%. The prevalence of diarrhea within the same time period was 13%. Prevalences at weaning (at the end of the grazing period) were: E. bovis 56.5%, E. zuernii 4.6%, C. parvum 3.7%, S. papillosus 14.4% and Trichostrogylida 84.5%. With the exception of E. zuernii, endoparasites were more frequently observed in healthy calves than in diarrheic calves. Weaning weights were available from 190 calves (33 herds). Statistical analyses of weaning weights revealed no evidence that an infection with helminths and/or protozoa within the first three months of life or at weaning had a negative influence on individual weight gain. PMID- 10481371 TI - [Salmonella infections in swine herds--epidemiology and importance for human diseases]. AB - Based on pertinent literature and research work performed by the authors, a report is presented on the presence of salmonellas in swine herds and the importance of these organisms as agents of disease in swine and source of infection for human salmonellosis. The share among human cases of salmonellosis which are caused by salmonellas originating from swine is estimated at ca. 20%. It has to be assumed that a very large proportion of swine herds is contaminated by Salmonella. Salmonellas may be introduced through infected pigs (parent animals, pigs from other herds added to the herd) or carriers among other animal species (e.g. rodent pests, birds) as well as by feeding stuffs with primary or secondary contamination. So far, the individual importance of the various routes cannot be reliably assessed. It appears that the level of Salmonella prevalence within a herd essentially depends on the hygienic conditions, the mode of keeping and the management of the animals. Serological examinations of meat juice permit conclusions as to the level of Salmonella contamination in slaughtered pigs. The results can thus be used for programmes to reduce the introduction of salmonellas into the food chain. PMID- 10481372 TI - [Animal nutrition in veterinary medicine--actual cases: copper intoxication in piglets]. AB - A case is described where piglets reacted with severely reduced feed intake, watery and dark faeces, poor growth and rough hair coat after delivery of a new charge of a commercial pelleted diet. Body temperature was normal, the behaviour appeared depressed. The diet contained 860 mg copper/kg, which was highly above the labelled concentration (160 mg/kg) and the concentration allowed by feed legislation (175 mg/kg for pigs up to 16 weeks of age). After changing to a new mixed feed the general condition of the piglets improved and faecal consistency and colour normalised within a few days. PMID- 10481373 TI - Elimination of tilmicosin in lactating ewes. AB - Tilmicosin was injected subcutaneously to lactating ewes once at a dose of 10 mg kg-1 b.wt. to determine its plasma, milk, urine and ruminal juice concentrations. Tilmicosin could be detected in all those fluids 30 minutes after injection. Milk and urine concentrations were higher than those of plasma and ruminal juice. The drug was detectable in milk, urine and plasma for 9, 4 and 3 days after injection, respectively. No amount of tilmicosin could be detected in ruminal juice 12 hours following administration. The mean peak concentration of tilmicosin in plasma and milk (Cmax) were 1.29 and 9.5 micrograms ml-1 and were obtained at (Tmax) 5.235 and 15.093 hours, respectively. The drug was slowly eliminated from plasma and milk as indicated by its long half-life (t1/2el) of 15.4 and 26.2 hours, respectively. The mean binding of tilmicosin to plasma and milk proteins in vitro was 16.8% and 26.8%, respectively. The drug was not bound to ruminal juice at any extent. The rate of tilmicosin renal clearance revealed that it was correspondingly increased with higher blood concentrations. While creatinine clearance showed no significant change after tilmicosin administration. The ratio (fractional clearance) between tilmicosin renal clearance to creatinine clearance was less than one indicating that the glomerular filtration is the main pathway of elimination through kidneys. The rate of ruminal gas fermentation in ewes was inhibited after subcutaneous injection of tilmicosin at a dose of 10 mg kg-1 b.wt. The tested samples taken at different time intervals from the rumen of ewes showed a subsequent reduction in the rate of fermentation as compared to control samples. The reduction was correspondingly increased with the increase of tilmicosin concentration in ruminal juice and returned to normal thereafter. PMID- 10481374 TI - Tissue residues of some sulphonamides in normal and Eimeria stiedai infected rabbits. AB - Tissue residues of sulphadiazine (SDZ), sulphadimidine (SDD) and sulphquinoxaline (SQ) were studied in healthy and E. stiedai infected rabbits following oral administration of 0.5 g/l drinking water for 5 days. The solid-phase extraction and HPLC was used to determine the concentration of the three sulphonamides in a single tissue sample. SDZ was detected in the liver and kidney in concentrations below the tolerance levels at day 5 and no residues could be detected at day 7 after drug withdrawal. SDD and SQ were detected in all of the tested organs of healthy rabbits up to day 5, where the highest concentration was reported in the liver (0.08 +/- 0.02 and 0.09 +/- 0.02 g/g respectively). In infected rabbits, the three sulphonamides were detected up to day 7 in concentrations higher than the tolerance limits (> 0.1 g/g) in the liver and kidney and lower levels in other tissues. A withdrawal period of 4 days for SDZ and 5 days for SDD and SQ in healthy rabbits and 7 days for SDZ and 8 days for SDD and SQ in E. stiedai infected rabbits is suggested. PMID- 10481375 TI - [Regulation of the contact sensitivity reaction by suppression of T gamma delta lymphocytes]. AB - Contact sensitivity (CS) is a classical example of in vivo T cell mediated immune response that is under regulation. It is well known that in normal mice suppression of CS can be mediated by T alpha beta cells tolerized by prior exposure to high dose of antigen (Ag). In this paper it was shown that treatment of defective TCR alpha -/- or TCR beta -/- mice with high dose of Ag may result in induction of T gamma delta suppressor cells, that are able to inhibit both adoptive cell transfer of CS in vivo and IFN-gamma production in vitro. These suppressor cells are characterized as: TCR gamma delta+, CD3+, CD4-, CD8-, CD28+, CD40L+, CD95 (Fas)+, Fc gamma R+ and NK1.1-. Suppression mediated by T gamma delta cells showed antigen specificity, but was not restricted by the MHC. T gamma delta suppressor cells express very strong down-regulatory activity where even 2.5 x 10(3) T gamma delta cells could suppress 7 x 10(7) CS-effector cells. Presented data may suggest that IL-4 released by T gamma delta suppressor cells is involved in the mechanism of their down-regulatory function. PMID- 10481376 TI - [Odontogenic tumors and neoplastic-like changes of the jaw bone. Clinical study and evaluation of treatment results]. AB - The purpose of the study was clinical analysis of 164 odontogenic tumours and tumour-like lesions of the jaws treated at the Department of Maxillofacial and Oral Surgery, Jagiellonian University College of Medicine between 1956 and 1996, and to evaluate early late and therapeutic results. Odontogenic tumours have a specific histological structure reflecting various stages of odontogenesis and are located mainly in the jaws, exceptionally in other parts of the skeletal system. Due to their specific structure and location they have been identified and classified by pathologists into a separate group of neoplasms differing from other tumours developing in the oral cavity and facial bones first of all in histogenesis, but also in biology, clinical manifestations and radiological sings. The currently recommended histological classification of odontogenic tumours established by the WHO in 1992 comprises a variety of odontogenic tumours and tumour-like lesions of bones of ectodermal, ecto-mesodermal and mesodermal origin as well as lesions histogenetically not associated with the dental organ, but constituting this group. They include histologically benign tumours, locally malignant and malignant tumours, the latter making up the least numerous group. Due to marked polymorphism histological diagnosis of odontogenic tumours is difficult, therefore co-operation between a clinicist, radiologist and pathologist is especially important. Treatment of choice in patients with odontogenic tumours is surgical operation: in case of benign tumours extirpation and curretage, whereas in case of malignant and locally malignant tumours- segmental resection of the jaws. Removal of extensive tumours is associated with the need of surgical tissue repair and prosthetic rehabilitation, and in young patients surgical treatment must be followed by orthodontic treatment. Clinical analysis included case histories of 164 patients admitted to the Department of Maxillofacial and Oral Surgery (Head: Professor MD S. B. Bartkowski) between 1956 and 1996. Additional control examinations were performed to analyze late results of therapy. The 40-year clinical material comprised 164 odontogenic tumours including 91 (55.5%) in women, and 73 (44.5%) in men. The mean age of the patients was 32.5 years, but about 30% of the patients were below 18. The tumour was most frequently located in the mandible i.e. in 118 (72.0%) patients, and in the maxilla in 44 (26.8%), and in both jaws in 2 (1.2%) patients. Due to tumour recurrence 19 (11.6%) patients were referred to our clinic from other centres, the remaining ones received their first treatment. The most frequent reason for patient referral was deformity of the jaws, maleruption or lack of teeth or their dislocation, and in case of extensive lesions face deformation and bulging of the tumour into the oral cavity. Radiological examination showed noncharacteristic transparencies, round or oval in shape, single- or multiventricular or foci of noncircumscribed bone densities usually in the vicinity of the teeth, frequently dislocated or unerupted. Rarely osteolysis of the roots of teeth or their amputation was found. Histological examination of the 164 tumours revealed (in order of frequency): ameloblastoma--60 (36.6%), central reparative giant cell granuloma--52 (31.7%), cementoma--15 (9.2%), odontoma--13 (7.9%), odontogenic myxoma--12 (7.3%), ameloblastic fibroma--4 (2.4%), calcifying epithelial odontogenic tumour--3 (1.8%), odontogenic fibroma--2 (1.2%), adenomatoid odontogenic tumour--2 (1.2%) and odontoameloblastoma--1 (0.6%). Of the 60 ameloblastomas three tumours were histologically malignant. Of the 164 patients 152 were selected for surgical treatment. Surgical treatment of histologically and clinically benign tumours included tumour extirpation, curettage, excision and/or electrocoagulation, whereas malignant and locally malignant tumours such as ameloblastomas, myxomas, Pindborg's tumours, invasive reparative giant c PMID- 10481377 TI - [Mechanism of action of neurotensin on microcirculation, metabolism and motility of the small intestine]. AB - The aim of this study was to investigate the influence of neurotensin on total microcirculatory blood flow, oxygen consumption, myoelectric activity and motility of the small bowel. The attempts were made to evaluate the role of L Arginine, NO-system in the mechanism of action of this peptide on microcirculation, metabolism and motor-activity of the intestine. The experiments were performed in two experimental models--dogs and rats. In Vetbutal anesthetised animals the systemic arterial pressure, superior mesenteric artery blood flow, microcirculatory blood flow and myoelectric activity of the small bowel were continuously monitored. In experiments with dogs effective capillary index, arterio-venous oxygen difference and oxygen consumption were analysed. Neurotensin administered intraarterially caused a dose depended increase in total and microcirculatory blood flow in the small bowel in both groups of animals. The experiment with dogs showed the rise of effective capillary index and intestinal oxygen consumption. Administration of neurotensin changed the profile of myoelectric activity of the small bowel similar to that of postprandial hyperaemia. Inhibition of NO-synthase by the intravenous administration of L-NNA reduced significantly the amplitude of neurotensin hyperaemia. Pretreatment with L-Arginine, the substrate of NO-synthase reversed the hemodynamic, but not motility effects of neurotensin. The results presented proved the role of the L Arginine-NO in circulatory mechanism, but not motility effects of neurotensin. PMID- 10481378 TI - Corticosterone response induced by intracerebroventricular administration of met enkephalin and naloxone in rats under stress. AB - In rats exposed to a 10 min restraint stress naloxone (0.1-10 micrograms) administered intracerebroventricularly (i.c.v.) did not substantially alter the hypothalamic-pituitary-adrenal (HPA) stress response, measured indirectly through serum corticosterone levels. Met-enkephalin (10-200 micrograms i.c.v.) dose dependently increased the stress-induced corticosterone response. The increase in corticosterone secretion elicited by met-enkephalin was markedly diminished by pretreatment with naloxone. The adrenergic alpha 1- and alpha 2-receptor antagonists, prazosin and yohimbine did not significantly affect the met enkephalin-induced increase in serum corticosterone levels in stressed rats. Conversely, propranolol, a beta-adrenergic receptor antagonist, abolished the met enkephalin induced corticosterone response. These results indicate that opioid receptors are not significantly involved in the HPA response to a short restrains stress. In stressed rats met-enkephalin stimulates, by a central mechanism, the hypothalamo-pituitary-adrenocortical activity via opioid- and beta-adrenergic but not alpha-adrenergic receptors. PMID- 10481379 TI - [Evaluation of autonomic nervous system function with heart rate variability analysis in patients with hyperthyroidism and during euthyroidism after pharmacologic and surgical treatment]. AB - The aim of the present study was both to estimate autonomic nervous system (ANS) function in patients with hyperthyroidism by the heart rate variability (HRV) analysis and to evaluate the impact of pharmacological and surgical treatment on the ANS function. Analysis of the HRV underwent 10 female patients in course of thyreotoxicosis and after reaching full clinical and biochemical euthyroidism, after pharmacological therapy and in month after surgical treatment. The 10 minutes records at rest, in horizontal position were evaluated. The HRV parameters like mean of the heart rate, mean of RR intervals, standard deviation of all normal RR intervals (SDNN), range of the heart rate variability, low frequency (LF), high frequency (HF) components of the heart rate power spectral density and LF/HF ratio were assessed. The results were compared to those obtained from 10 age-, sex-, and body mass index-matched control subjects. The statistical significance (p < 0.05) was found in reduction of range of RR intervals, in increase of LF/HF ratio and in decrease of SDNN in hyperthyroidism in comparison to the control group (151.6/346.8 ms; 2.4/0.74; 24.4/57.2 ms2). In course of pharmacological euthyroidism there were statistically significant (p < 0.05) increase of range of RR intervals, reduction of LF/HF ratio and increase of SDNN in comparison to hyperthyroidism (270/151.6 ms; 0.995/2.4; 39/24.4 ms2). In euthyroidism after surgical treatment all the above parameters kept the similar levels as in pharmacological euthyroidism (no statistical significance for p < 0.05). On the base of the outcomes it was considered that in hyperthyroid patients there is advantage of sympathetic part of ANS over parasympathetic one which is due to sharp reduction of parasympathetic system activity. Pharmacological therapy with thyreostatics normalises balance of ANS to the level of the control group and after surgical treatment the balance keeps the same. Moreover, in the estimation of ANS as important as LF/HF ratio is the mean range of RR intervals. PMID- 10481380 TI - Effect of naloxone on central adrenergic stimulation of corticosterone secretion. AB - The interaction of central opioid with adrenergic system in stimulation of the hypothalamic-pituitary-adrenal (HPA) axis was investigated in conscious rats. All the tested drugs were administered intracerebroventricularly (i.c.v.), naloxone 15 min before the adrenergic agonists. Phenylephrine (30 micrograms), clonidine (10 micrograms) and isoprenaline (10 micrograms), the alpha 1-, alpha 2- and beta adrenergic agonists as well as noradrenaline (10 micrograms) and adrenaline (10 micrograms) significantly increased the serum corticosterone levels, measured 1 h after drug administration. Naloxone (0.01-1 microgram) did not markedly influence the corticosterone response to phenylephrine and isoprenaline and diminished by a half the response to clonidine. Naloxone also significantly decreased the corticosterone response to noradrenaline but did not substantially alter the response to adrenaline. Noradrenaline potently stimulates the HPA axis via alpha 2-adrenoceptors, whereas adrenaline involves mainly alpha 1- and beta-adrenergic receptors. These results indicate that central opioid system is significantly involved in mediating the pituitary-adrenocortical response to clonidine and noradrenaline but not phenylephrine, isoprenaline and adrenaline. This suggests an interaction of opioids at central alpha 2- but not alpha 1-receptor sites. PMID- 10481381 TI - [Autonomic nervous system dysfunction in Parkinson's disease evaluated by the heart rhythm variability test]. AB - It is well recognized that autonomic dysfunction are common in Parkinson's disease (PD). Fourteen patients with early PD and 8 patients with advanced PD aged from 38-71 were investigated. Heart rate variability at rest differ significantly between patients with advanced PD and age-matched controls. Cardiovascular autonomic dysfunction in PD mainly affects parasympathetic but also sympathetic system, and occurs only in advanced cases. Heart rate variability is a useful non-invasive test to assess autonomic dysfunction in PD. PMID- 10481382 TI - [Hepatoprotective effect of flower pollen lipid extract in paracetamol-induced hepatotoxicity in mice]. AB - Hepatoprotective effects of the lipid flower pollen extract (LEPK) in paracetamol intoxication in mice were shown. Normalisation of A1AT and LDH--biochemical indicators of necrotic changes in the hepatic cells, and high protection ultrastructural cell organelle such as mitochondrion substantially testifies to the hepatoprotective effect of investigated lipid flower pollen extract on the hepatic cells. PMID- 10481383 TI - [The level of anti-HBs in blood serum of persons selected from a group of Krakow inhabitants]. AB - The aim of the study was to determine levels of anti-HBs antibodies in blood of 1490 adults coming from Cracow province. The first group of 1451 people did not suffer from viral hepatitis B in the past and were never vaccinated against hepatitis B; 86.2% of them did not demonstrate any protective level of anti-HBs antibodies, but 13.8%--presented antibody level equal to 10 IU/L or higher. The second group consisted of 14 individuals who did not suffer from virus hepatitis and who were vaccinated 1-5 years before examination (basic vaccination of three doses); they all demonstrated the protective anti-HBs antibody level (> or = 10 IU/L). In the third group there were 20 subjects who were post-hepatitis convalescents (they demonstrated symptomatic acute hepatitis 2-4 years before examination) and were never vaccinated; 7 of them did not present any protective level of anti-HBs antibodies, but the remaining 13 individuals demonstrated certain immunity status. The last group of 5 people, who suffered from acute hepatitis in the past and had received incomplete immunization (1 dose) previously demonstrated antibody levels of differentiated protective value. PMID- 10481384 TI - [Use of flow cytometry in evaluation of cellular changes in interstitial lung diseases]. AB - Flow cytometry (FC) usefulness for pulmonary reactive cell changes examination in interstitial lung diseases was assessed. Bronchoalveolar lavage (BAL) lymphocyte direct (two-color and three-color) phenotyping was carried out in 63 patients with sarcoidosis (subdivided according to the disease radiological stage and smoking status), 23 patients with systemic sclerosis (SSc) and 12 individuals exposed to silica dusts. 33 healthy volunteers were used as controls. Routine BAL cytology and peripheral blood lymphocyte (PBL) typing was performed in all tested subjects. FC results quality control was performed according to BD Simultest IMK Plus criteria (for PBL) modified by the author. The conventional lymphocytic gate (PBL light scatter based) was excluded, as the source of error in L-BAL subset analysis. Two-color L-BAL phenotyping and alveolar lymphocyte CD45FITC/SSC gating was found to be usually sufficient. However, in poor-lymphocyte materials (e.g. in healthy smokers) three-color method, using anti-CD45 PECy5 monoclonal antibody for staining each patient sample, should be considered. No significant changes in lymphocyte subsets were found in FC as compared with alkaline phosphatase immunocytochemical method. The prevalence of CD3+ cells with only a few lymphocytes expressing NK (CD3-CD16 or 56+), T suppressor (CD8+ 11b+) or B cell phenotype--as compared with PBL, was found in BAL of all examined subjects. CD8+ cell subset was dominated by T cytotoxic cell phenotype (CD8+ CD11b-) in all tested groups. L-BAL T cells expressed sensitized memory cell phenotype--CD4+ (CD8+)CD45RO+ in sarcoidosis, SSc and controls (the staining not performed in silica exposed persons). Characteristic BAL cytological and immunological pattern (lymphocytic alveolitis, increased CD4/CD8, high CD3(CD4)+HLA-DR+ percentage) was observed in pulmonary sarcoidosis. SSc patients showed reduced BAL CD4/CD8 ratio, due to T cytotoxic cell predominance (the changes were more distinct in the group with clinical pulmonary changes). Early activation marker (CD25) was expressed on the elevated percentage of BAL T helper cells in SSc. High CD4+CD25+ cell percentage was the most characteristic sign of lymphocyte activation in individuals exposed to silica. Summing up, the reference values for L-BAL subsets were proposed. CD4/CD8 ratio alterations in interstitial lung disorders should be interpreted as the local imbalance between T helper memory cells and sensitized T cytotoxic lymphocytes. FC characterization of alveolar lymphocytes provides insight into the pathogenesis of pulmonary diseases. PMID- 10481385 TI - [Toxic effect of chemical pollution in lower and upper silesia on morpho-chemical differentiation of human placentae]. AB - Monitoring of the environmental chemical pollution in southern Poland from 1994 to 1996 involved histochemical studies of 71 human term placentae collected in the Low Silesian Legnica-Glogow Copper Region and in Upper Silesian Industrial Region. Control placentae were obtained from weakly polluted eastern Carpathian regions (Bieszczady). The studies were aimed at comparing the effect of regions of long and short industrialization periods on structure and enzyme activities of placentae taken from healthy non-smoking women. The chemical pollution of both the industrial environments decreased the oxidative enzyme activity, particularly that of cytochrome c oxidase in the villous trophoblast. This resulted in a compensatory augmented number of placental villi below 30.5 microns in diameter which expanded the mother-child exchange area and in a decreased proportion of larger villi. The statistically significant compensatory changes in villous proportions were more pronounced in the recently industrialized Low Silesian region. In all Silesian placentae abundance of mineral deposits was found. The Silesian neonates showed a noteworthy though statistically insignificant birth weight decrease. PMID- 10481386 TI - [Environment and health--studies using biomarkers]. AB - Present day society is questioning more and more the effect of the evolution of modern agriculture on the quality of life. Two new tools, biomarkers and satellite images, may be used to provide a better understanding of the links which exist between the quality of the environment, the food chain and human health. This multi-disciplinary approach has for some years now joined together diverse specialists from INSERM, the CNRS and the University. Experiments are undertaken in real conditions on several sites of large scale farming land in the north of the Paris Basin. The aim is to study the quantitative and qualitative transfers of pollutants travelling in soils and waters towards agricultural food products, and then to man. The first results of laboratory analysis, concerning the use certain genes of thermic shock proteins HSP 70 and metallothionein MT2A as biomarkers of disequilibrium, are very conclusive. Furthermore, the connection of these genes with a fluorescent (reporter) gene and found in farmed vegetation, could enable the use of satellite images, and this possibility seems to be very promising. The ultimate goal is to improve knowledge of the direct effects of modern agricultural practices on human health and to participate in the debate about the place of modern agriculture in the development of modern society. PMID- 10481387 TI - Is the use of catecholamine before ischemic arrest safe? Effect of catecholamine on rat heart ischemia/reperfusion injury. AB - Using an isolated working heart model, we studied the effects of dopamine, adrenaline, or noradrenaline pretreatment on ischemia/reperfusion injury. Hearts from Wistar rats were perfused in the first 20-minute working mode, 15 minutes in Langendorff mode, and in the second 20-minute working mode. Hearts were treated with dopamine (0.52 and 2.60 mmol/L), adrenaline (16 and 80 nmol/L), or noradrenaline (16 and 80 nmol/L) during the second working perfusion, then arrested with St. Thomas' Hospital cardioplegic solution and subjected to global ischemia (37 degrees C or 20 degrees C). During reperfusion, recoveries of cardiac function and creatine kinase leakage were measured. At 37 degrees C, dopamine and adrenaline had a harmful effect at both doses; noradrenaline was harmful at a high dose but beneficial at a low dose. At 20 degrees C, adrenaline, dopamine, and noradrenaline had a harmful effect at high doses but no harmful effect at low doses. To determine the role of beta adrenergic stimulation before ischemia, a dose-response study was undertaken with isoprotelenol and milrinone at 37 degrees C. Combined pretreatment with isoprotelenol and milrinone accelerated ischemia/reperfusion injury dose-dependently. Preischemic beta adrenergic stimulation thus plays a significant role in the deleterious effect of catecholamine pretreatment at high doses. At low doses, however, the effect of the inotropic agent could be changed depending on ischemic temperature. Our results suggest that catecholamine should not be given at high doses before ischemia, regardless of temperature during ischemia. PMID- 10481388 TI - Results of surgery for primary lung cancer based on the new international staging system. AB - OBJECTIVES: This study clarified the results of surgery for primary lung cancer based on the new international staging system. BACKGROUND: On December 1997, the Japan Lung Cancer Society adopted a new TNM staging system which had already received international recognition. SUBJECTS AND METHODS: The subjects of this study were 1062 consecutive previously untreated patients who underwent pulmonary resection for primary non-small cell lung cancer between January 1975 and December 1992. RESULTS: The postoperative 5-year survival rate for all patients was 58.5%. Pathological staging demonstrated a survival rate which was 73.2% in stage I, 46.8% in stage II, 26.7% in stage III, and 20.0% in stage IV. In the staging subgroups, the survival rate was 79.6% in stage IA, 62.4% in stage IB, 62.2% in stage IIA, 42.0% in stage IIB, 26.9% in stage IIIA, and 26.3% in stage IIIB. Concerning the pm patients, the survival rate was 20.2% in pm1 and 20.0% in pm2, while the survival rate of the patients with N0 was 45.7% in pm1 and 40.0% in pm2. CONCLUSIONS: A significant difference in the 5-year survival rate was recognized between the new stages IA and IB, and between the new stages IIA and IIB. When pm patients are diagnosed without lymph node metastasis, the opportunity for resection should not be lost. PMID- 10481389 TI - Indications and operative techniques for combined aortoesophageal resection. AB - Combined aortoesophageal resection was performed in 8 patients, including 7 with esophageal carcinoma and 1 with aortoesophageal fistula. Aortic resection procedures included segmental resection with permanent aorto-aortic bypass (1 case), segmental resection with graft interposition (1 case), semicircumferential resection with patch aortoplasty (3 cases), wedge resection with lateral aortorrhaphy (1 case), and resection of adventitia (2 cases). Protective methods during aortic cross-clamping included one aorto-aortic permanent bypass, one subclavian-aortic bypass, and three axillo-femoral bypass. Postoperative complications include mediastinal abscess, paresis, arrythmia, and pneumonia. Five patients with esophageal carcinoma died within 6 postoperative months. In 4 of these 5 nonsurvivors, metastasis to distant organs including the liver, bone and peritoneal cavity were found at the time of death or autopsy. Those early recurrence cases were characterized by skip lesions and extensive lymph node metastasis with extranodal invasion. The clinical benefit of aortoesophageal resection will be attained by careful preoperative evaluation for case selection and a sufficient protective method for aortic cross-clamping. PMID- 10481390 TI - Use of arterial grafts for coronary revascularization. Experience of 2987 anastomoses. AB - OBJECTIVES: The aim of this study was to evaluate the performance and the quality of arterial grafts for coronary artery bypass grafting at The Heart Institute of Japan, Tokyo Women's Medical University. PATIENTS AND METHODS: From January 1970 to March 1998, 2987 arterial grafts, including left and right internal thoracic arteries, gastroepiploic artery, radial artery and inferior epigastric artery, were used in 1673 patients. In the same period, 1225 saphenous vein grafts were used. Early graft patency was angiographically determined. Also, histological evaluation of operative specimens and preoperative angiographic evaluation of arterial grafts were performed. RESULTS: The total number of hospital deaths was 38 (2.3%). Of 4212 grafts, 3919 grafts (93%) were evaluated angiographically and 3714 of 3919 grafts were patent (94.8%). The patency rate of internal thoracic artery was better than that of gastroepiploic artery (p < 0.0001), radial artery (p = 0.0005) and saphenous vein grafts (p < 0.0001). However, the patency rate of gastroepiploic artery was better than that of saphenous vein grafts (p = 0.04), while no significant difference was detected between gastroepiploic artery and rdial artery. Only one internal thoracic artery specimen obtained at surgery showed atherosclerotic change, but all gastroepiploic artery specimens had moderate to severe atherosclerotic changes with CD68-positive cell infiltration. Only one patient's left internal thoracic artery out of 200 was not angiographically useable as a conduit, while multiple stenotic lesions in gastroepiploic artery were observed. CONCLUSIONS: In the graft selection for CABG, the primary choice is internal thoracic artery and the secondary choice is right internal thoracic artery, from the standpoint of histological and angiographic evaluation, gastroepiploic artery and/or radial artery, depending on the target anastomotic site, degree of stenosis, and in situ or free use is the third choice. PMID- 10481391 TI - Arterial revascularization. 18-year experience with coronary artery bypass grafting in familial hypercholesterolemia. AB - Familial hypercholesterolemia is characterized by a high plasma level of cholesterol and is frequently associated with rapidly progressing coronary heart disease. The internal thoracic artery is recognized as the conduit of choice for coronary artery bypass grafting. This study was performed to determine whether multiple arterial grafting was associated or not with additional benefits for patients with familial hypercholesterolemia. Between June 1980 and March 1998, 95 patients with familial hypercholesterolemia. underwent a total of 103 coronary artery bypass procedures with one hospital death. The patients were divided into 3 groups according to the type of bypass graft. Group 1 included 31 patients with only saphenous vein grafts; Group 2,48 patients with one arterial graft and supplemental vein grafts; and Group 3, 24 patients with multiple arterial grafts. The overall actuarial survival rate was 90.9% at 10 years and 81.8% at 18 years. The overall actuarial freedom from recurrent angina was 68.9% at 10 years and 55.8% at 16 years. The actuarial survival rate in group 2 was higher than that in Group 1 (p < 0.05). There was no difference in the actuarial survival or in the freedom from cardiac events between Group 2 and Group 3. Single arterial grafting improved the long-term survival in patients with familial hypercholesterolemia. However, no additional benefit from multiple arterial grafting was identified. PMID- 10481392 TI - Surgical management of idiopathic tracheal stenosis. AB - Most nonmalignant upper tracheal stenoses are caused by prolonged endotracheal intubation or tracheostomy, and idiopathic stenosis is uncommon. A 43-year-old woman complained of increasing shortness of breath during exercise over a year prior to admission. She had no significant past medical history, including endotracheal intubation. Bronchoscopy and tracheal tomography revealed nonmalignant circumferential upper tracheal stenosis 20 mm long. Single-stage surgical resection with cricotracheal anastomosis completely relieved her respiratory symptoms. Idiopathic tracheal stenosis is extremely rare, and the treatment of choice is surgery. PMID- 10481393 TI - Coronary artery aneurysm after stent implantation. AB - A coronary artery aneurysm that developed 6 months after coronary stent implantation in the left anterior descending artery, was treated by double coronary artery bypass grafting due to restenosis of the affected vessel and progression of right coronary artery stenosis. Although the poststenting aneurysm initially remained postoperative angiography showed that it had disappeared. In addition to thrombotic occlusion, another mechanism behind its disappearance may be that geometric changes of the implanted stent caused by heart retraction during surgery closed the entrance to the aneurysm. PMID- 10481394 TI - Treatment of mediastinitis arising after replacement of the ascending aorta. AB - Mediastinitis due to graft infection is a serious and potentially lethal complication associated with replacement of the ascending aorta. We present the case of a 67-year-old man with this condition for the aneurysm and chronic dissection. Mediastinitis and sepsis were diagnosed and debridement, irrigation with povidone solution and omental transposition were performed successfully. Continuous closed irrigation prior to omental transposition without replacement of the infected graft is useful for treating mediastinitis after ascending aortic or arch replacement. PMID- 10481395 TI - Thromboembolism originated from the pulmonary artery stump after Fontan operation. AB - Cerebral thromboembolism is a rare but serious complication after Fontan operation. This is the report of a patient who underwent a successful intracardiac thrombectomy for cerebral thromboembolism after Fontan operation. A 2-year-old girl was referred to us with the diagnosis of tricuspid atresia without pulmonary stenosis, normally related great arteries, and a ventricular septal defect. Although she underwent a successful Fontan operation and division of the main pulmonary artery, she developed a cerebrovascular event at 3 weeks after the operation. Echocardiography demonstrated a large thrombus within the residue of the main pulmonary artery, and suggested that the thrombus had migrated into the systemic circulation by way of the ventricular septal defect. At 2 weeks after the cerebrovascular event, she underwent thrombectomy and excision of the pulmonary valve. Although she has developed slight left-sided hemiparesis, she is leading a normal life at 1 year after the operation. PMID- 10481396 TI - Atrial septal defect as an uncommon cardiovascular malformation with Turner's syndrome. AB - Cardiovascular malformations are frequently observed in Turner's syndrome. Bicuspid aortic valve and coarctation of aorta are commonly associated with Turner's syndrome while an atrial septal defect is unusual. Here report a rare case of atrial septal defect with Turner's syndrome. PMID- 10481397 TI - [Detection of anti-Chlamydia trachomatis antibody by means of enzyme immunoassay using synthetic peptide antigen]. AB - Newly developed diagnostic kits for the detection of Anti-Chlamydia trachomatis, Peptide-Chlamvdia (LOY: Meiji Milk Products Co., Ltd., Tokyo; for IgG and IgA), were evaluated using the microimmunofluorescence assay (MIF) as the gold standard. These results were also compared to results of testing by Sero-IPALISA and immunoblot (I-B). Detection by LOY in based on enzyme immunoassay with synthetic peptides as the antigen. Thirty serum samples from pediatric patients and 130 serum samples from gynecology patients were used. All 26 pediatric samples that were positive for Chlamydia pneumoniae IgG antibody tested negative with LOY, indicating that the presence of the antibody against C. pneumoniae did not affect the assay by LOY. For 90 gynecological samples, the total, the positive and the negative agreement rates for IgG were quite high; i.e. 87.8%, 90.0% and 70.0% (LOY vs MIF), 85.6%, 85.0% and 90.0% (Sero-IPALISA vs MIF), and 92.0%, 94.9% and 70.0% (I-B vs MIF), respectively. On the other hand, many cases of MIF (-) and LOY (+) discrepancy were seen in IgA detection. In order to better understand the basis for such disagreement. 34 serum samples were collected from patients whose cervical samples were negative for the Chlamydia group antigen based on the assay with IDEIA-Chlamydia. They were then assayed by MIF and LOY. The total, the positive and the negative agreement rates for IgG were 91.2%, 100% and 90.9%, while the total and the negative agreement rates for IgA were 88.2% and 88.2% (there were no IgA positive cases). Furthermore, 6 serum samples (1 case of MIF (+) LOY (+) and 5 cases of MIF (-) LOY (+)) were provided to determine whether LOY detects C. trachomatis specific IgA antibody. Increasing amounts of C. trachomatis serovar L2 were added to the serum samples resulting in a progressive decrease in their reactivity in the LOY assay. These results lead us to speculate that LOY can reveal even low levels of C. trachomatis specific IgA antibody. In conclusion, LOY can be used as an useful kit for detecting C. trachomatis antibody. PMID- 10481398 TI - [Pulmonary complications in adult measles]. AB - The clinical features of pulmonary complications were evaluated in 15 patients with adult measles diagnosed by clinical and serologic investigations. Of those 15 patients, 12 were admitted because of high fever, sever anorexia and dyspnea. Almost all patients were found to have thrombocytopenia, elevations of LDH and aminotransferase. Pulmonary infiltrates were present in only 2 of the 12 patients (16.7%) on whom chest roentgenograms were performed, but hypoxemia (PaO2 < 70 Torr) were present in 8 of the 12 patients (66.7%). Pulmonary function tests in 8 patients showed mild decrease in VC, moderate decrease in FEV1 and severe decrease in V25. These findings suggest that hypoxemia in patients with normal chest radiographs may be largely caused by bronchiolitis. The observations of sequential peak flow rate measurement showed that severe pulmonary dysfunction continued for 4-5 days after the onset of the rash. To avoid the development of respiratory failure, patients with measles should be carefully monitored for bacterial superinfections of the respiratory tract especially within several days after the appearance of the rash. PMID- 10481399 TI - [Clinicobacteriological studies on the nine cases with upper genital tract Mycoplasma hominis infection]. AB - Clinicobacteriological characteristics of nine cases isolated Mycoplasma hominis from the genital tract were studied, and the following results were obtained: elevation of IgG antibodies to M. hominis was measured by ELISA in all cases, but in the MI method only one case showed an elevation of metabolic inhibitory antibody. Convalescent sera from seven patients showed additional and high density bands which were not recognized by acute phase sera in immunoblotting. It was thought that in two patients M. hominis was a causal bacteria for pelvic inflammatory disease (PID). In three cases, it was suggested that M. hominis was related to a premature delivery and idiopathic labor. As infectious symptoms, two patients had body temperatures of more than 38 degrees C but other cases showed 37-37.8 degrees C. Though all cases showed an elevation of CRP, six elevations were slight. As a medication beta-lactam agents were administrated, but their efficacy was not recognized. Furthermore, two patients showed spontaneous recovery in spite of improper antimicrobial agents administration or drainage combined with antimicrobial agents. From the above results. It was thought that M. hominis played a causative role in upper genital tract infection. PMID- 10481400 TI - [Background factors and clinical course of patients with urosepsis]. AB - In this study, we analyzed background factors and clinical courses of 31 patients having 32 episodes of urosepsis, which was defined as the disease when the same species of bacteria were simultaneously isolated both from urine and blood in a febrile (> or = 38 degrees C) patient. Most of the patients had a malignant disease. These patients were classified into 3 groups by febrile patterns, based on [responder], [partial responder] and [non-responder], [non-responder] was one in which fever persisted for more than 7 days. Background factors such as those of bacteria and host were compared among the three groups. Pseudomonas aeruginosa was more frequently isolated from both urine and blood of patients having not only the [responder] but also the [non-responder]. Patients having the [responder] and [partial responder] had a higher frequency of isolation of Escherichia coli (E. coli) from urine and blood, suggesting that urosepsis caused by E. coli may respond more easily to antimicrobial chemotherapy. While the direct cause of urosepsis was not identified in 6 patients, manipulations of the urinary tract were a direct cause of the disease in all 26 other patients. An indwelling catheter in the central vein or urinary tract alone did not influence the clinical courses of patients. However, when patients with respiratory or renal dysfunction, or a cerebrovascular accident had an indwelling catheter, the urosepsis was intractable for antibacterial chemotherapy. Antimicrobial chemotherapy using a single agent was effective in all patients having the [responder]. Combination chemotherapy or alternation of agents was eventually needed for treatment in more than half of the patients with the [partial responder] or [non-responder]. No patients died from the disease. PMID- 10481401 TI - [Analysis of tuberculin reaction of tuberculous children below 4 years of age]. AB - To clarify whether the size of tuberculin reaction could be used as an useful index of the severity of tuberculosis, we analyzed the sizes of tuberculin reaction (TR) of 60 children below 4 years of age with active tuberculosis at the time of diagnosis. Of 60 patients, 53 (88.9%) had positive reactions to tuberculin. The mean size of TR of 60 patients was 24.0 +/- 13.9 mm and maximum size was 60 mm. Seven patients who had no reaction to the tuberculin skin test consisted of three primary complex and four serious tuberculosis (two miliary tuberculosis and two tuberculous meningitis). The patients without BCG vaccination showed significantly smaller TR than the patients with BCG vaccination (p < 0.05). The patients less than 1 year of age showed significantly smaller TR than the patients of 4 years of age (p < 0.05). The patients with serious tuberculosis showed significantly smaller TR than the patients with primary complex (p < 0.05). Of patients with primary complex, there were no difference of the size of TR between the patients with pulmonary tuberculosis (III) and hilar lymphadenopathy (H). Together with, it did not necessarily mean that negative TR showed no infection with tuberculosis and the sizes of TR depended on the severity of tuberculosis in infantis and young children. PMID- 10481402 TI - [Influence of dexamethasone on the clinical course of bacterial meningitis in children. Especially on secondary fever. Experiences in 27 institutions]. AB - Of pediatric patients with purulent meningitis seen at the institutions listed in the title page of this paper between 1986 and 1994, 93 patients treated with antibiotics and dexamethasone (DXM) were compared with 91 patients treated with antibiotics alone. The patients receiving antibiotics with dexamethasone achieved overall improvement in inflammatory symptoms and signs and cerebrospinal fluid findings and became afebrile significantly earlier than those receiving antibiotics alone. However, some of the patients became febrile again. The secondary fever rate for the DXM group was much higher than that for the antibiotic alone group (p < 0.0001). In most of the rebounded cases, the body temperature rose above 38 degrees C and remained elevated for 2-4 days. Cerebrospinal fluid (CSF) was cultured daily in 54 and 32 patients receiving antibiotics with and without DXM, respectively. Although this study was not a controlled study in a strict sense, these patients compared. In both groups, the CSF became mostly culture-negative within 48 hours. In a few patients receiving DXM, however, it became culture-negative after 72 hours or longer. DXM caused an adverse effect in a patient with meningitis caused by Streptococcus pneumoniae. The adverse effect was mild gastrointestinal bleeding, which recovered spontaneously. From the findings described above, the use of DXM combined with antibiotic therapy was considered to accelerate the relief from fever and improvement of inflammatory symptoms and signs and CSF findings. The body temperature rose again in more than half of the patients receiving DXM, but fell to normal spontaneously without treatment. The elevation doubtlessly could not be distinguished from recurrence of the meningitis itself or complications. It seems to be likely that no treatment but careful observation is required even if the fever recurs as far as the CSF findings showed favorable progress with excelluent general conditions. When DXM is given, it is essential that CSF tests and culture are repeated during the early stages and the progress is monitored carefully. PMID- 10481403 TI - [Clinical evaluation of anaerobic infections in patients with bronchopulmonary infections diagnosed by transtracheal aspiration]. AB - We evaluated the clinical and bacteriologic features in the patients with bronchopulmonary infections isolated anaerobes from transtracheal aspirates between April 1990 and March 1998. Some anaerobe was isolated in 42 (10.9%) in 387 patients whom we performed transtracheal aspiration (TTA), in 42 (15.7%) of 268 in whom some organism was isolated from TTA, or in 42 (16.3%) of 257 patients in whom some bacterium excluding acid-fast bacteria, fungi or mycoplasma from TTA. The isolation rate of anaerobic bacteria was 93.3% in the patients with lung abscess, 22.7% in the patients with nosocomial pneumonia, 19.4% in the patients with community-acquired pneumonia, 26.7% in the patients with acute exacerbation of chronic lower respiratory tract infection (CLRTI), 1.6% in the patients with persistent infection of CLRTI, and 3.0% in the patients with acute bronchitis, respectively. The major anaerobes, isolated from TTA, were Peptostreptococcus micros and Prevotella melaninogenica. The aerobic bacteria were isolated with anaerobic bacteria in 32 of 42 patients at the same time. The quantitive grade of colonial growth of anaerobes was equal to or more than aerobes in the patients with lung abscess and pneumonia. We mostly administrated 3rd generation cephems or carbapenems with or without clindamycin for the treatment of anaerobic infections. Forty-one of 42 patients were cured only by the therapy of antimicrobial agents, but pneumonia patient with lung cancer died in spite of adequate antimicrobial therapy. These results suggest that the anaerobic infections are important in the bronchopulmonary infections. PMID- 10481404 TI - [Evaluation of a new screening assay kit for the combined detection of HIV p24 antigen and antibody--comparison of the performance of the new kit and HIV antibody assay kits]. AB - DUO is an automated HIV infection screening test kit based on the combined detection of p24 Ag and anti-HIV-1 and anti-HIV-2 IgG in human sera or plasma using the ELFA technique (Enzyme-Linked Fluorescent Assay). The performance of DUO was compared with that of HIV-1/HIV-2 3rd generation EIA plus and particle agglutination (PA) test. A total of 141 seropositive sera, 3 seroconversion panels, 300 seronegative sera and 387 potentially cross-reactive serum samples were tested. One hundred and forty one seropositive sera in Japan and Cameroon were all positive with DUO. Three seroconversion panels (panel Q, Z, AE) were tested to evaluate sensitivity. In Panel Q, infecution was detected seven days earlier with DUO than with the 3rd generation EIA plus and PA. In Panel AE, infection was detected four days earlier with DUO than with the single antibody assays. Three hundred seronegative sera from Kanagawa prefectural public health centers were all negative with DUO as well as PA test. Three hundred and eighty seven potentially cross-reacting samples were tested to challenge the specificity of the assay. These included samples from pregnant women and hepatitis patients. In four of the 204 samples from pregnant women, false-positive results were observed with DUO. In three of the 183 samples from hepatitis patients, false positive results were also obtained with DUO. All samples of 7 DUO positive results were negative with western blot test. Five of them were negative with RT PCR and 2 of them were not tested because there were not enough samples. Thirty cross-reacting (false-positive) samples by PA test from blood donors were tested by DUO, and all of these were negative by DUO. PMID- 10481405 TI - [Cost of medication for influenza infected elderly inpatients]. AB - Influenza vaccine is recommended for the elderly, a high-risk group for influenza infection. Unlike in many developed countries, the rate of influenza vaccination is extremely low in Japan. One of the primary reasons for this low level of vaccination use may be insufficient study on the cost-effectiveness of influenza vaccination in Japan. We determined the cost of medical tests and medications for inpatients with influenza in a referred hospital. We compared the medical costs by (1) level of daily life activity, (2) presence of pneumonia, and (3) living/dead status, 9 months after the infection. In addition, we set up a control group of patients for comparison of their medical costs to those of the influenza patients. Mean costs were 37,279 (+/- 26,784) yen for patients, and 2,361 (+/- 4,893) yen for controls. Mean costs were 32,424 (+/- 30,935) yen for inpatients without limitations to activity, 44,075 (+/- 20,937) yen for bed-bound inpatients, 44,614 (+/- 28,609) yen for inpatients with pneumonia, 27,009 (+/- 22,783) yen for inpatients without pneumonia, 57,624 (+/- 21,041) yen for inpatients who died within 9 months of the onset of influenza, and 16,934 (+/- 11,920) yen for inpatients who were alive 9 month after influenza infection. PMID- 10481406 TI - [Antibacterial effects of cacao mass on enterohemorrhagic Escherichia coli O157:H7]. AB - The antimicrobial activities of aqueous cacao mass extract against enterohemorrhagic Escherichia coli (EHEC) O157:H7 006 strain were studied. Hot water extract of cacao mass (cocoa extract) was shown to inhibit the growth of EHEC O157:H7 006 strain in PBS or CAYE medium. In addition, the production of verotoxins (types 1 and 2) of EHEC O157:H7 006 strain was significantly inhibited by 8.0% cocoa extract. The cocoa extract did not neutralize the cytotoxity of verotoxins, but had inhibitory effect on adhesion of verotoxins to the target Vero cells. These results demonstrate that cacao mass has antimicrobial effects on EHEC O157:H7. PMID- 10481407 TI - [Granular lymphocyte-proliferative disorder following initiation of protease inhibitor therapy in a patient with acquired immunodeficiency syndrome]. AB - The authors experienced granular lymphocyte-proliferative disorder (GLPD) with increased lymphocyte and CD4 cell counts after HIV treatment with a protease inhibitor indinavir in a 51-year-old male AIDS patient. GLPD proved to be the chronic type, but we could not differentiate whether it was the T-cell type or the NK-cell type. EB virus was found to be activated and its chronic activity was suggested. We should note in this case that EB virus was involved in the GLPD onset and that GLPD followed the treatment with a protease inhibitor. PMID- 10481408 TI - [Atypical Shigella sonnei strains isolated overseas travelers]. PMID- 10481409 TI - [Outbreak of multidrug-resistant Mycobacterium tuberculosis infection in the middle and advanced age]. AB - We experienced a small outbreak of multidrug-resistant Mycobacterium tuberculosis infection (MDR-TB) among persons of in the middle and advanced age. The index case was 48-year-old man, and had complained productive cough since January 1996. He visited a doctor due to his symptom, and chest X-ray revealed cavitary lesion and sputum smear was positive for tubercle bacilli. He could not continue his admission because of his absence without leave and drinking, he was discharged on Day 54. The drug resistance was observed for INH (0.1 microgram/ml), RFP, and SM. Later, case 2, 52-year-old male, and case 3, 43-year-old-male, who were companions in mah-jongg with the index case, were diagnosed as pulmonary tuberculosis. The analysis of restriction fragment length polymorphism (RFLP) was done on 3 strains, and all showed the same pattern. Among other companions in mah jongg with the index case, case 4, 28-year-old male, was treated as MDR-TB, and the drug resistance pattern was the same to that of the index case, but the details were unknown. Case 5, 65-year-old male, was diagnosed as drug sensitive pulmonary tuberculosis, thus he might incidentally suffer from pulmonary tuberculosis at the same time. Case 6, 46-year-old male, who had been treated for alcoholic liver cirrhosis, was introduced to our hospital as his sputum smear was positive, and the drug resistance pattern was observed similar to that of the index case. All the companions in mah-jongg suffered from MDR-TB except case 5. The RFLP analysis showed that the index case, case 2, and case 3 were caused by the same strain of M. tuberculosis. The drug resistance pattern of, case 4 and case 6 was the same to that of the index case. Based on these findings, it is highly suspected that this small outbreak was originated from the index case. PMID- 10481410 TI - [Epidemiological study of microepidemics of tuberculosis by restriction fragment length polymorphism analysis]. AB - Restriction fragment length polymorphism (RFLP) analysis was performed with 13 events considered as microepidemics of tuberculosis occurred in Osaka prefecture from July 1995 to July 1998. In 7 out of 13 events, isolates from patients involved in each event showed identical RFLP patterns, hence these events were verified as microepidemics. Out of 7 microepidemics, three were intrafamily infection, the other three occurred in work places and the remaining one was in a school occurred among students. The total number of patients who were involved in microepidemics was 19 and their ages distributed in the wide range from 18 to 69 years old, but 11 were at twenties or forties. The total delay in case-finding was 2 months in the shortest case, and more than 14 months in the longest. In the remaining 6 of 13 events, the isolates from patients were showed different RFLP patterns, although they were suspected as microepidemics. Furthermore, in the 2 microepidemics, one isolate showed different RFLP pattern from the other isolates involved in each events. These facts suggest that there could be many overlooked sources of tuberculosis infection in Osaka. PMID- 10481411 TI - [Effects of secretory leukocyte protease inhibitor on the production of some cytokines and nitric oxide by murine peritoneal macrophages in response to lipopolysaccharide stimulation and M. avium complex infection]. AB - Secretory leukocyte protease inhibitor (SLPI), a serine protease inhibitor abundantly found in mucous secretions of lung, is thought to serve as an important protective component in the secretory fluids at sites of degenerative and inflammatory diseases. In this study, we examined the effects of SLPI on the production of a proinflammatory cytokine, TNF-alpha, and immunosupressive cytokines, IL-10 and transforming growth factor-beta (TGF-beta) by macrophages (M phi s), in response to lipopolysaccharide (LPS)-mediated stimulation and M. avium complex (MAC) infection, using recombinant half-sized SLPI (1/2 SLPI) consisting of C-terminal domain (Arg55-Ala107) of intact SLPI. In addition, effects of SLPI on the production of nitric oxide radicals (NO), important antimicrobial effectors of M phi s against micro-organisms, by these M phi s were studied. First, when the number of TNF-alpha producing cells in the LPS-stimulated M phi population was counted by the ELISPOT assay, more than half of the M phi s acquired TNF-alpha secreting ability at 24 hr after LPS stimulation. On the other hand, MAC-infected M phi s produced detectable amounts of TNF-alpha into culture fluid during the first 24 hr. In both cases, 1/2 SLPI did not affect the LPS- or MAC-induced TNF-alpha production by M phi s. Second, when the production of IL-10 and TGF-beta by M phi s was determined by measuring the amounts of these cytokines accumulated in M phi culture fluids by ELISA, the following was observed. M phi IL-10 production was rapidly increased in the early phase of cultivation after LPS stimulation or MAC infection, peaking on day 1 and thereafter declining to normal level by day 14. Half-sized SLPI did not affect IL 10 production of LPS-stimulated M phi s, while it caused slight enhancement of IL 10 production by MAC-infected M phi s. M phi TGF-beta production was initiated in the middle phase (day 7) of M phi cultivation and continued until day 14. Notably, 1/2 SLPI markedly potentiated the TGF-beta producing ability of the LPS stimulated M phi s. Moreover, 1/2 SLPI caused moderate increase in the TGF-beta production by MAC-infected M phi s. Third, significantly potentiated NO production was observed in M phi s during the first 2 days after LPS stimulation and MAC infection. Half-sized SLPI did not affect the NO production by LPS stimulated or MAC-infected M phi s. These findings indicate that SLPI up regulates the production of some immunoregulatory cytokines including IL-10 and TGF-beta, particularly the latter, by M phi s in response to LPS stimulation or MAC infection, thereby suggesting the possibility that SLPI may exhibit antiinflammatory effects in vivo, especially patients with bacterial infections including MAC diseases, through the modulation of M phi expression of some immunosuppressive cytokines. PMID- 10481412 TI - [MICs and MBCs of levofloxacin, clarithromycin, and KRM-1648 for Mycobacterium tuberculosis and M. avium complex residing in MONO-MAC-6 human macrophage-like cell and A-549 human type II alveolar epithelial cell lines]. AB - In this study, we determined the MICs and MBCs of levofloxacin (LVFX), clarithromycin (CAM), and KRM-1648 (KRM) for Mycobacterium tuberculosis (MTB) strain Kurono and M. avium complex (MAC) strain N-444 residing in MONO-MAC-6 human macrophage-like cells (MM6-M phi s) and A-549 human type II alveolar epithelial cells (A-549 cells). First, the MICs of LVFX for MTB replicating in MM6-M phi s (1 microgram/ml) and A-549 cells (2 micrograms/ml) were 4 to 8 times higher than its MICs for extracellular MTB growing in 7HSF medium. In contrast, the MICs of CAM for intracellular MTB residing in both the cells (2-4 micrograms/ml) were 4 to 8 times less than its MICs for extracellular MTB organisms. On the other hand, the MICs of KRM for extracellular MTB were nearly the same as its MICs for intracellular MTB residing in both types of the cells. Secondly, the MICs of LVFX and CAM for extracellular MAC were not significantly different from their MICs for intracellular MAC residing in MM6-M phi s and A-549 cells. The MIC of KRM for MAC residing in A-549 cells was 0.25 microgram/ml, and this value was 32 times higher than its MIC for MAC residing in MM6-M phi s (0.008 microgram/ml). Thirdly, the MBCs of test drugs for intracellular MTB and MAC residing in both types of the cells were somewhat longer than their MBCs for extracellular organisms. These findings indicate that, in the case of pulmonary infections with MTB or MAC, the therapeutic efficacy of a given drug, especially KRM, is more or less influenced by the bacterial location in the host lung tissues where the mycobacterial pathogens survive and multiply, i.e., either alveolar M phi s, type II alveolar epithelial cells, or surrounding environment. PMID- 10481413 TI - [Two cases of duodenal tuberculosis]. AB - The present study described 2 cases of patients with duodenal tuberculosis. Case 1 was a 55 year-old Japanese male patient with pulmonary tuberculosis and past medical history of subtotal gastrectomy (Billroth II reconstruction). Tubercle bacilli were positive both on smear and culture from his sputum and feces. Because of complaining right hypochondralgia, upper gastrointestinal endoscopy was performed and revealed multiple ringed ulcers in the afferent limb of duodenum. Histopathological study of biopsy specimen demonstrated granulomatous inflammation as well as acid-fast bacilli confirmed by Ziehl-Neelsen staining. Tissue culture was positive for M. tuberculosis. Colonic tuberculosis was demonstrated by barium enema. Case 2 was a 45 year-old male patient with pulmonary tuberculosis in association with severe uncontrolled diabetes mellitus. Sputum polymerase chain reaction test was positive for M. tuberculosis. Granulomatous inflammation and positive acid-fast bacilli in biopsy specimen obtained from ulcers in the descending portion of the duodenum made a diagnosis of duodenal tuberculosis. No other intestinal tuberculous lesion was recognized. Since 1988, 11 cases of duodenal tuberculosis including the presented two cases have been reported in Japan. Most of the recent cases had tuberculous lesions in the descending portion of the duodenum and were diagnosed as duodenal tuberculosis by endoscopic examinations, while the majority of the cases reported before 1987 had tuberculosis in the more distant portions of the duodenum and were diagnosed mainly by surgical procedures. PMID- 10481414 TI - [Assessment of oxygen transport in individual splanchnic organs using critical oxygen extraction]. AB - Vulnerability in individual organs under ischemic stress can be assessed by critical oxygen extraction (critical O2ER). We measured critical O2ER in the whole-body, total splanchnic organs, liver and gut using a biphasic regression model of the oxygen delivery-consumption relationship in 7 dogs subjected to hemorrhagic stress under 1% isoflurane anesthesia. The difference in critical O2ER between the whole-body and total splanchnic organs was not significant, while hepatic critical O2ER was significantly larger than that of the gut (P < 0.05), 89.7 +/- 2.4% (mean +/- SD) and 66.1 +/- 5.1%, respectively. This finding indicated that the liver functions well under ischemic conditions due to its excellent ability to extract oxygen. The anatomical character of the sinusoid and the microcirculatory improving effect of isoflurane may affect the ability of oxygen extraction of the liver. The critical threshold of hepatic venous oxygen saturation as a marker of hepatic oxygen imbalance was considered to be below 10%. PMID- 10481415 TI - [Influence of PEEP on left ventricular diastolic function]. AB - We investigated the influence of PEEP on left ventricular diastolic function using the pressure-volume loops by employing a micromanometer catheter and a conductance catheter placed in the left ventricle of the eight mongrel dogs. Time constant of diastolic interval was shortened under the condition of 15 cmH2O of PEEP compared with the control value (PEEP 0 cmH2O) and the diastolic stiffness constant was prolonged under 15 cmH2O of PEEP. It was considered that the shortened time constant of diastolic interval was due to the decrease of preload. The prolongation of diastolic stiffness constant was possibly due to compression of the heart by increased intrapleural pressure. We conclude that decreased compliance of left ventricle caused by PEEP might impair the left ventricular function. PMID- 10481416 TI - [The enhancement of formalin induced agitation behavior by intrathecal administration of prostaglandin E1]. AB - The spinal action of pro-inflammatory agent, prostaglandin E1 (PGE1) was investigated in rats, using formalin test. Sprague-Dawley rats were implanted with chronic intrathecal catheters. PGE1 0.25 microgram or 2.5 micrograms injected intrathecally. Ten minutes later, 5% formalin 50 microliters was injected to the dorsum of one hindpaw. For the time-response analysis, the total number of flinches were counted and expressed as responses per minute for each rat. For the dose response analysis, the cumulative responses per minute over the first 5 min (phase 1) and the interval 10-60 min (phase 2) following the formalin injection were calculated for each rat. PGE1 2.5 micrograms produced a significant increase of flinching in both phase 1 and phase 2. Dose dependent increase of flinching was seen only in the phase 2. Touch evoked allodynia was observed after the injection of PGE1 2.5 micrograms, which lasted about 50 minutes. The enhanced flinches induced by intrathecal PGE1 were antagonized by the coadministration of MK-801, hemoglobin or methylene blue. Allodynia was antagonized only by coadministration of methylene blue. These results indicate that hyperalgesia induced by spinal PGE1 in both phase 1 and phase 2 of the formalin test involves an increased release of glutamate and the activation of NMDA receptor in the spinal cord. The mechanism of allodynia observed in the rats given high dose of PGE1 differs from that of hyperalgesia. PMID- 10481417 TI - [Effects of speed of injection on anesthesia induction with propofol and fentanyl]. AB - We examined the effects of injection rate of propofol on injection pain and postinduction hypotension and bradycardia when fentanyl was administrated before propofol. Fifty-five patients premedicated with midazolam and atropine were randomly allocated to two groups. Three minutes after administration of fentanyl 100 micrograms, propofol 1.5 mg.kg-1 was injected to a forearm vein at a rate of 800 ml.hr-1 in Group FS or 1 ml.s-1 in Group FR. Anesthesia was maintained with 67% nitrous oxide in oxygen and supplemental propofol infusion. The incidence and severity of pain on injection were not influenced with injection speed. The rapid rate of injection significantly shortened the induction time. The decrease in systolic and diastolic blood pressures and heart rate after induction were not affected by injection speed. In conclusion, rapid injection of propofol after fentanyl was effective to shorten the induction time without increasing the postinduction hypotension and bradycardia. PMID- 10481418 TI - [Effects of the bolus injection rate on anesthesia induction with propofol]. AB - We examined the influence of the bolus injection rate of propofol on the cardiovascular depression and injection pain. Fifty-one patients of ASA grade 1 or 2 were randomly allocated to two groups. After premedication with midazolam 0.06 mg.kg-1 and atropine 0.006 mg.kg-1 i.m., propofol 2 mg.kg-1 was injected to a forearm vein at a rate of 800 ml.hr-1 in Group A or 1 ml.s-1 in Group B. Anesthesia was maintained thereafter with 67% nitrous oxide in oxygen and propofol infused at 6 mg.kg-1.hr-1. The incidence and severity of pain on injection were lower in Group B compared with Group A, but the difference was not statistically significant. The induction time was significantly shorter in Group B than in Group A (40 vs. 73 sec: P < 0.01). There were no significant differences between the two injection rates in peak reductions in systolic and diastolic blood pressure and heart rate. In conclusion, rapid injection of propofol was effective to shorten the induction time without any adverse effects. PMID- 10481419 TI - [Anesthetic management for carotid and coronary artery surgery--concomitant versus two stage operation]. AB - We reviewed the anesthetic management of 16 patients with concomitant severe coronary and carotid artery diseases. Eight patients underwent concomitant operations of coronary artery bypass graft and carotid endarterectomy, while the other 8 patients underwent two stage operation. Candidates for concomitant operations had unstable angina or serious coronary disease such as three vessel disease or severe stenosis of LMT. In comparison, most of patients undergoing two stage operation had symptomatic or occlusive carotid disease. In all cases, anesthesia was maintained with fentanyl and midazolam and the perfusion pressure during cardiopulmonary bypass was maintained above 70 mmHg. Some patients received thiopental or propofol for brain protection. The concomitant operations required much more transfusion and longer operation time than two stage operation. In addition, several cases of the concomitant operation needed intra aortic balloon pumping or high dose of catecholamines. Indications for concomitant operation or two stage operation have to be determined through discussion among anesthesiologist, neurovascular as well as cardiovascular surgeons. PMID- 10481420 TI - [Premixing of 5% dextrose in Ringer's acetate solution with propofol reduces incidence and severity of pain on propofol injection]. AB - The purpose of this study was to compare the effect of premixed 5% dextrose in Ringer's acetate solution and premixed lidocaine with propofpl on the reduction of pain during injection of propofol in adult patients. We conducted a prospective, randomized, double-blinded trial. Ninety-six patients were randomly allocated to one of three groups according to the agents added to 1% propofol 20 ml; Group C, normal saline 2 ml, Group L, 2% lidocaine 2 ml, and Group A, 5% dextrose in Ringer's acetate solution 2 ml. The pain on injection was rated as none, mild, moderate, or severe. Seventy percent of patients in the C group experienced pain, while 33% and 25% of patients experienced pain in the A group and the L group, respectively. Forty-two percent of patients in the C group complained moderate to severe pain but only one patient in both A group and L group. In conclusion, 5% dextrose in Ringer's acetate solution premixed with 200 mg propofol significantly reduces incidence and severity of pain associated with propofol injection and is easier to use than premixed lidocaine. PMID- 10481421 TI - [Cardiovascular responses during laryngeal mask airway insertion in normotensive, hypertensive and chronic renal failure patients]. AB - The hemodynamic response to the insertion of the laryngeal mask airway (LM) following induction with propofol 2 mg.kg-1 was assessed and compared in normotensive (Normal), hypertensive (HT) and chronic renal failure (CRF) patients (n = 23 in each group). Before induction, in HT and CRF groups blood pressure and rate pressure products (RPP) were higher than in Normal group (P < 0.05). Although blood pressure and RPP were decreased in every patient by induction with propofol, no patients needed vasopressor drugs. The decreases of blood pressure and RPP were larger in HT and CRF groups than in Normal group (P < 0.05). There were no differences between groups in heart rate and rate of successful LM insertion. We concluded that LM insertion with propofol 2 mg.kg-1 was an effective induction method preventing the adverse circulatory responses in normotensive, hypertensive and chronic renal failure patients. PMID- 10481423 TI - [The changes of peripheral skin temperature in elderly patients under spinal anesthesia]. AB - The purpose of this study was to evaluate the changes of body temperature in elderly compared with younger patients during spinal anesthesia. Twenty six patients, ASA I-II were divided into two groups, (13; under 30 years of age, 13; above 60 years) who received spinal anesthesia with 0.3% dibucaine at the interspace between L 3 and L 4. All patients were anesthetized below T 8 level. Central temperature was measured at the forehead, and peripheral skin temperature was measured at the finger tip and the toe tip for 25 minutes at every 5 minutes. In both groups, forehead temperature and finger tip skin temperature were unchanged, but toe tip skin temperature increased 5 minutes after spinal anesthesia. The degree of toe tip skin temperature change was significantly lower (P < 0.01) in elderly patients and the speed was also slower. We recognized that the change in peripheral skin temperature during spinal anesthesia is not different between elderly and younger patients, but the speed of temperature change is slower in elderly patients. PMID- 10481424 TI - [Anesthetic management for newborn pharyngeal teratoma]. AB - Epignathus (pharyngeal teratoma) is a rare disease of newborns associated with a high mortality secondary to airway obstruction in the neonatal period. We report anesthetic management of a newborn with epignathus who underwent tumor resection. He was delivered vaginally at 39 weeks of gestation and Apgar scores were 9 at 1 and 5 min. The tumor originated from the palate, almost filled the oral cavity and protruded through the mouth with its external part 6 x 7 cm in size. He could breathe with the head and mass turned to the left. The excision of the tumor was scheduled on the fifth day of life. Mask ventilation and laryngoscopy were considered impossible. Fiberoptic nasal intubation was successfully performed with topical anesthesia without sedation. Tumor was resected with blood loss of 103 gm. The trachea was extubated on the third postoperative day and the postoperative course was uneventful. For safe management of cases of pharyngeal teratoma, careful preoperative assessment of the airway is most important and sufficient preparation and careful intubation are mandatory to keep airway patent. The perioperative bleeding from the tumor and the airway obstruction by the tumor or its remnant after the excision could also be hazardous to the airway. PMID- 10481422 TI - [Caesarean section in a patient with a history of HELLP syndrome]. AB - A 36-year-old woman underwent emergency caesarean section following the diagnosis of HELLP syndrome. Four years earlier, after having undergone the same procedure for HELLP syndrome, she had experienced hypovolemic shock, renal failure, and disseminated intravascular coagulopathy during the postoperative period. This time, the patient showed bleeding, elevation of liver enzymes (ALT, AST, LDH) and a reduction of antithrombin III activity in the 36th week of pregnancy. Anesthesia was induced by thiamylal 4 mg.kg-1 and suxamethonium 1 mg.kg-1 and after delivery maintained by oxygen-nitrous oxide-isoflurane, and all procedures were performed without any incident. No major complications such as intraperitoneal bleeding, renal failure, or disseminated intravascular coagulopathy occurred during the postoperative period. It is suggested that caesarean section should be carried out as soon as possible after the diagnosis of HELLP syndrome is confirmed. PMID- 10481425 TI - [Anesthetic management of a patient with hypohidrotic ectodermal dysplasia]. AB - Hypohidrotic ectodermal dysplasia (HED) is a rare congenital anomaly complex characterized by hypodontia, hypotricosis and hypohidrosis. There have been only a few reports of anesthetic management of patients with HED. We managed a 20-year old man with HED, who underwent debridement and skin grafting under epidural anesthesia, without untoward events. Potential problems in anesthetic management of patients with HED are also discussed. PMID- 10481426 TI - [Seasonal variation in the incidence of acute aortic dissection in Yokohama]. AB - In order to determine any possible influence of climatic variables on the incidence of acute aortic dissection (AAD), we reviewed all patients with a diagnosis of AAD treated at Yokohama City University Urafune Hospital between January 1994 and December 1997 retrospectively. One hundred and eight patients with AAD were admitted into the hospital during the period. Forty-two patients (39%) developed AAD during the winter, when the lowest temperature was below 6 degrees C. Thirty-one patients (29%) developed AAD in May or October, when the lowest temperature was almost 15 degrees C. We suspect that the incidence of AAD was influenced not only by the atmospheric temperature but also by other factors, for example, the change of the atmospheric pressure. PMID- 10481427 TI - [A case of pulmonary edema following upper airway obstruction after general anesthesia]. AB - A 30-year-old man underwent tonsillectomy and laryngomicrosurgery under nitrous oxide oxygen-isoflurane anesthesia. Preoperative physical examinations and interview revealed no cardiopulmonary abnormalities. Two minutes after extubation, he showed dyspnea with marked inspiratory efforts and cyanosis due to laryngeal spasm. The SpO2 decreased from 100% to 80%. He was reintubated after administration of suxamethonium 100 mg and ventilated with 100% oxygen. At that time pink frothy sputum came out from his airway. He was diagnosed as pulmonary edema from analysis of arterial blood gases and chest X-ray. He received mechanical ventilation with positive end-expiratory pressure in the ICU. Twenty hours later the pulmonary edema was improved and he was extubated uneventfully. He was discharged from the hospital on the 8th post-operative day. We reported a case of pulmonary edema after laryngeal spasm. It was suggested that a patient after acute upper airway obstruction should be carefully treated considering secondary pulmonary edema. PMID- 10481428 TI - [Continuous total intravenous anesthesia is recommended for wake-up test]. AB - Total intravenous anesthesia (TIVA) has been recommended in view of avoiding air pollution. However, intermittent administration of anesthetic agents has a large disadvantage of delayed emergence time. We reported that continuous TIVA with propofol, ketamine, vecuronium and buprenorphine (PKBp) could bring rapid emergence time. An 8-year old female with spinal scoliosis underwent spinal instrumentation under continuous TIVA. Wake-up test was done twice during surgery smoothly after stopping infusion of anesthetic agents in the cocktail. We conclude that continuous TIVA is one of the most recommended anesthetic methods for wake-up test. PMID- 10481429 TI - [Propofol anesthesia for an emergent caesarean section in a patient with asthma]. AB - We report a case of emergency caesarean section due to bleeding from placenta praevia under general anesthesia in a patient with asthma. General anesthesia was induced by propofol 150 mg and suxamethonium 80 mg, and operation was started immediately after tracheal intubation under cricoid cartilage pressure. While induction-delivery time took about 270 seconds, Apgar scores at 1 and 5 minutes are 8 and 9 points, respectively. Plasma concentration of propofol in umbilical vein was 0.73 microgram.ml-1 by HPLC. Although anesthetic maintenance was carried out by oxygen-nitrous oxide-isoflurane after delivery, no asthmatic attack was seen throughout the operation. Anesthetic induction by propofol for emergency caesarean section might be safe and useful in a patient with asthma with little effects to neonates. PMID- 10481430 TI - [Automated anesthesia billing by electronic anesthesia record keeping system]. AB - There has been loss of anesthesia fee by errors of filling out billing sheets manually. A large loss of anesthesia fee was pointed out by the audit during the past several years. In order to prevent these billing error, we have developed an automated anesthesia billing system combined with an electronic anesthesia record keeping (EARK) system. The system derives all the anesthesia cost-related parameters from the EARK database and calculates anesthesia fee according to the logic of the Japanese health care insurance system. After implementing the system, anesthesiologists and circulating nurses became free from filling out billing sheets. The accuracy of the automated billing was proved by the comparison with hand filled sheets. The survey showed that the system prevented 2.5 million yen of billing loss in a month. Such an economic impact of the system proves the rationale of an EARK as a cost containment tool. PMID- 10481431 TI - [A new bite block also serving as an endotracheal tube holder for infants]. AB - We developed a new type of bite block with a combined function as an endotracheal tube (ETT) holder for infants and small children to prevent airway troubles caused by tube kinking, dislodging, extubation and oral membrane trauma. One mm thick plastic plate sized 3.5 x 2 cm was curved to make an open roll. The outer surface of the roll was covered and glued with soft plastic tube (5.0 mm ID endotracheal tube), cut in 3.5 cm length to give an elastic outer surface for the patient's comfort. The rolled ends were diagonally cut to make an oblique slit of 3 mm width. A t-shaped flange made of soft vinyl plate was fixed at a third of the length of the roll to maintain the block's position relative to the lips and to make the fixation of the tube easier. In practical use, after endotracheal intubation is performed as usual, this bite block is put into the mouth and positioned at the oral angle with the flange on the patient's skin. The ETT is fit into the slit of the roll. The skin-facing surface of the flange is pasted to the skin with the double stick material usually used for colostomy stoma. The ETT and the bite block are fixed en bloc with fixing tapes around the mouth. Our bite block has following advantages over other types of bite blocks and tube holders especially for children; 1) the volume of foreign bodies (ETT and bite block) occupying the oral cavity can be reduced and this attenuates the patient's discomfort, 2) good holding of the ETT can prevent its dislodging and decrease the incidence of accidental extubation and 3) suctioning is easier because of wide oral space. The four sizes of the bite block suitable for 4.0, 4.5, 5.0, 5.5 and 6.0 mm ID ETTs are manufactured. We applied this device to several ICU patients and found its use practical and safe. PMID- 10481432 TI - [Progress in the therapy of rheumatism]. PMID- 10481433 TI - [Epibatidine. An old work principle rediscovered]. PMID- 10481434 TI - [Sildenafil (Viagra). Hormonal properties, indications and legal problems]. PMID- 10481435 TI - [Bleeding from the otorhinolaryngeal (ORL) area]. PMID- 10481436 TI - Proliferative potential of malignant glioma cells before and after interstitial brachytherapy. AB - The viability of tumor cells in radionecrotic tissue after interstitial brachytherapy (BRTX) was evaluated using immunohistochemical markers of proliferative potential in primary and recurrent tumors. Tumor specimens from 30 patients with malignant gliomas (14 anaplastic astrocytomas, 16 glioblastomas) taken before and after BRTX were examined using MIB-1 monoclonal antibody. Histological examination of specimens obtained by craniotomy or stereotactic biopsy after BRTX revealed tumor recurrence in 18 patients and radionecrosis in 12 patients including two with pure radionecrosis and 10 with a mixture of both tumor and radionecrosis. The MIB-1 index of the tumors with radionecrosis was 7.6 +/- 5.5%, and that of the primary tumors was 17.0 +/- 11.2%, showing a significant difference (p < 0.05). There was no significant difference between the MIB-1 index of the primary tumors with local recurrence after BRTX and the primary tumors which underwent radionecrosis. Although morphologically viable tumor cells were found in the radionecrotic tissue, BRTX causes a reduction in the proliferative potential of these tumor cells. PMID- 10481437 TI - Long-term results of ventrolateral thalamotomy for patients with Parkinson's disease. AB - This study evaluated the long-term outcome for 53 patients with idiopathic Parkinson's disease treated by stereotactic thalamotomy between 1977 and 1996 at our institute. Significant reduction of tremor and rigidity of the contralateral extremities persisted throughout the follow-up period (mean 8.8 years) in 44 patients who underwent unilateral thalamotomy. These effects resulted in postoperative improvement of activity of daily life (ADL) with reduced dosage of levodopa. The effect of surgery on akinesia was limited and postoperative progression of akinesia was related to the postoperative deterioration of ADL. Multivariate analysis disclosed that the preoperative akinesia score was the critical factor for poor outcome. Nine patients underwent bilateral thalamotomies at a mean interval of 56 months. Five patients were obviously benefited from the second thalamotomy. The only perioperative complication was large intracerebral hematoma at the lesion site in one patient. This study confirmed the reliable and persistent effect of thalamotomy. Patients with Parkinson's disease whose disability is mainly caused by tremor and/or rigidity will be benefited from this procedure. Second thalamotomy, contralateral to the initial side, may be indicated if the ADL deteriorates due to the progression of the symptoms on the non-treated side. Patients disabled by advanced akinesia are not good candidates for thalamotomy. PMID- 10481438 TI - Avulsion fracture of the anterior half of the foramen magnum involving the bilateral occipital condyles and the inferior clivus--case report. AB - A 38-year-old male presented with an avulsion fracture of the anterior half of the foramen magnum due to a traffic accident. He had palsy of the bilateral VI, left IX, and left X cranial nerves, weakness of his left upper extremity, and crossed sensory loss. He was treated conservatively and placed in a halo brace for 16 weeks. After immobilization, swallowing, hoarseness, and left upper extremity weakness improved. Hyperextension with a rotatory component probably resulted in strain in the tectorial membrane and alar ligaments, resulting in avulsion fracture at the sites of attachment, the bilateral occipital condyles and the inferior portion of the clivus. Conservative treatment is probably optimum even for this unusual and severe type of occipital condyle fracture. PMID- 10481439 TI - Variant arteriovenous fistula of the superior sagittal sinus--case report. AB - A 57-year-old male presented with a rare variant of dural arteriovenous fistula, located in the wall of an unobstructed superior sagittal sinus. Drainage occurred through a cortical vein no longer connected to its parent sinus, which filled up a cluster of transmedullary running veins, one of which was the presumed site of hemorrhage. Arterial blood was supplied via the external carotid artery branches. This type of fistula seriously increases the risk of hemorrhage in the patient and therefore requires complete obliteration. Attempts to embolize the fistula failed. The draining vein was isolated and coagulated resulting in permanent occlusion of the fistula. The fistula probably developed through a process of thrombophlebitis and revascularization via arterioles of the vein rather than previous occlusion of the sinus. PMID- 10481440 TI - Bilateral chronic subdural hematomas resulting in unilateral oculomotor nerve paresis and brain stem symptoms after operation--case report. AB - An 85-year-old male presented with bilateral chronic subdural hematomas (CSDHs) resulting in unilateral oculomotor nerve paresis and brainstem symptoms immediately after removal of both hematomas in a single operation. Initial computed tomography on admission demonstrated marked thick bilateral hematomas buckling the brain parenchyma with a minimal midline shift. Almost simultaneous removal of the hematomas was performed with the left side was decompressed first with a time difference of at most 2 minutes. However, the patient developed right oculomotor nerve paresis, left hemiparesis, and consciousness disturbance after the operation. The relatively marked increase in pressure on the right side may have caused transient unilateral brain stem compression and herniation of unilateral medial temporal lobe during the short time between the right and left procedures. Another factor was the vulnerability of the oculomotor nerve resulting from posterior replacement of the brain stem and stretching of the oculomotor nerves as seen on sagittal magnetic resonance (MR) images. Axial MR images obtained at the same time demonstrated medial deflection of the distal oculomotor nerve after crossing the posterior cerebral artery, which indicates previous transient compression of the nerve and the brain stem. Gradual and symmetrical decompression without time lag is recommended for the treatment of huge bilateral CSDHs. PMID- 10481441 TI - Persistent primitive hypoglossal artery associated with cerebral aneurysm and cervical internal carotid artery stenosis--case report. AB - A 71-year-old female had vertigo attacks once or twice a day secondary to vertebrobasilar insufficiency. Left carotid angiography revealed persistent primitive hypoglossal artery (PPHA) associated with a large internal carotid artery (ICA) aneurysm and severe stenosis of the ICA. The bilateral vertebral arteries were hypoplastic. The basilar artery was opacified via the PPHA but not via vertebral arteries. Clipping of the aneurysm was performed first because the risk of rupture of the aneurysm was not negligible. One month after clipping, carotid endarterectomy using a T-shaped shunt system was successfully performed. The postoperative course was uneventful and the vertebrobasilar ischemic attacks did not recur. Left carotid angiography demonstrated complete obliteration of the aneurysm and disappearance of the carotid artery stenosis. Low ICA flow (70 ml/min) and low stump pressure of the PPHA (25 mmHg) strongly suggested low perfusion of the posterior circulation. Carotid endarterectomy may be essential for augmentation of the posterior circulation in patients with PPHA associated with ICA stenosis. PMID- 10481442 TI - Progressive myelopathy caused by dural arteriovenous fistula at the craniocervical junction--case report. AB - A 68-year-old male presented an unusual dural arteriovenous fistula (AVF) located at the craniocervical junction. Magnetic resonance imaging revealed dilated perimedullary veins around the spinal cord at C-1 and C-2 levels, as well as high intensity signals in the spinal cord on T2-weighted images. Vertebral angiography identified an AVF at the point where the right vertebral artery penetrates the dura. The fistula was a single and direct communication between the vertebral artery and the spinal vein. Surgical interruption of the fistula at its venous side resulted in prompt improvement of both motor and sensory signs and symptoms. PMID- 10481444 TI - Galea-cranium suture method for prevention of the subcutaneous fluid collection- technical note. AB - A simple method is described for preventing cerebrospinal fluid (CSF) from collecting in the subcutaneous space following a supratentorial osteoplastic craniotomy. A few pairs of oblique holes through which suture materials may pass are made by drilling in the surface cortex of the bone flap. The galea aponeurotica is sutured to the bone flap by means of these holes. These galea cranium sutures reduce the subgaleal free space in which pooling of CSF or hematoma may occur. PMID- 10481443 TI - Ehlers-Danlos syndrome associated with multiple spinal meningeal cysts--case report. AB - A 40-year-old female with Ehlers-Danlos syndrome was admitted because of a large pelvic mass. Radiological examination revealed multiple spinal meningeal cysts. The first operation through a laminectomy revealed that the cysts originated from dilated dural sleeves containing nerve roots. Packing of dilated sleeves was inadequate. Finally the cysts were oversewed through a laparotomy. The cysts were reduced, but the postoperative course was complicated by poor wound healing and diffuse muscle atrophy. Ehlers-Danlos syndrome associated with spinal cysts may be best treated by endoscopic surgery. PMID- 10481445 TI - The suture applicator for replacing a bone flap--technical note. AB - A new instrument was developed for passing and holding sutures during the replacement of a bone flap. The new device is a simple straight aluminum shaft, 6 cm in length and 0.7 mm in diameter with a groove on both ends for holding the suture. The shaft can be easily bent with the fingers to attain the desired curve and more suitable manipulation. Passing the shaft through the straight hole in the cranium or the cranial flap was very easy and convenient. No dural damage occurred during 80 procedures using this shaft. No needle holder or forceps for temporary clipping sutures were necessary, which reduced the operative time and mental and physical burden on the operator and the nurses. The mean time per hole with our instrument (45.8 +/- 9.2 sec) was significantly shorter compared to conventional methods with a circular needle, needle holder, and many forceps for temporary clamping of sutures (65.6 +/- 13.2 sec). PMID- 10481446 TI - [Leprous neuropathy--observation from the standpoint vasculitis]. AB - Pathogenesis of leprous neuropathy was studied from the standpoint of vasculitis. In vasculitic syndrome, mononeuritis multiplex is the most specific peripheral nerve injury. It occurs most frequently in small vessel vasculitis. Among them, the incidence is highest in microscopic polyangitis (MPO), Churg-Strauss Syndrome (CSS), and Wegener's granulomatosis (WG). All of them are categorized as ANCA associated disease. Because the disease activity is parallel with ANCA titer and ANCA is related directory to the pathogenesis of necrotizing vasculitis as well as the occurrence of peripheral neuropathy. Other small vessel vasculitis, such as lupus or rheumatoid vasculitis which are believed to occur by immune complex mechanism, are accompanied seldom by peripheral neuropathy. In WG, there are granulomatous lesions in upper and lower respiratory tracts which are supposed to be induced by Th1 dominant CD4 T cell infiltration. In cases of leprosy, if peripheral neuropathy were related to small vessel vasculitis, there might be two different pathogenesis. One is granulomatous vasculitis like WG in tuberculoid type. The other is immune complex type necrotizing vasculitis like SLE in lepromatous type, especially ENL. The site of these events may be small vessel in epineurium. Further immunologic studies are needed to clarify detailed mechanisms of leprous neuropathy. PMID- 10481447 TI - [Hansen's Disease Control Program on move in Japan]. PMID- 10481448 TI - Experimental studies on peripheral nerve repair: a possibility of application to cure nerve complication of Hansen's disease. PMID- 10481449 TI - [The control leprous peripheral neuropathy and chemotherapy]. AB - Clarithromycin(CAM), Roxithromycin(RXM), Minocycline(MINO) and Fosfomycin(FOM) has anti-inflammatory action and immunomodulatory activity, while the anti mycobacterium leprae activity is shown. CAM and RXM suppress the rat carrageenin edema, and MINO suppresses the rat adjvant arthritis. There is the immunosuppression on adrenocorticosteroid while the inflammatory cytokine is suppressed. CAM, MINO, FOM suppresses the inflammatory cytokine, while it has the immunomodulatory activity. Fusidic acid(FA) suppresses the inflammatory cytokine with the action of being similar to cyclosporin A, and it has the immunomodulatory activity. New macrolides derivatives, CAM and RXM showed the inflammatory regulation, and MINO showed the anti-inflammatory activity with FA. The combination chemotherapy can be enforced, while peripheral neuropathy is prevented by the control of the leprosy reaction. PMID- 10481450 TI - [Global leprosy, current status and a future outlook]. AB - Successful "Leprosy Elimination Programme" since 1991 managed to reduce global case load to nearly 1/10 in 10 years. However, this rapid fall of case detection/incident rate. This means that even after year 2,000, control effort of leprosy as an infectious disease must be sustained, while adequate control/care of leprosy as a deformity/disability causing disease need more attention. PMID- 10481451 TI - [Histopathological changes of peripheral nerves and blood vessels in Hansen's disease]. PMID- 10481452 TI - [Bacteria killing by macrophages via NF-IL6 gene dependent mechanism: the susceptibility to Mycobacterium leprae in NF-IL6 knockout mice]. AB - Transcription factor, NF-IL6 recognizes the same nucleotide sequences as C/EBP, and it is predominantly expressed in macrophages. Tanaka et al. reported that NF IL6 knockout mice are highly susceptible to Listeria monocytogenes and Salmonella typhimurium due to impairment of bacteria killing by activated macrophages. We have tried to see the susceptibility for Mycobacterium leprae infection with intraperitoneal(i.p.) or both hind foot pad (BHF) in the NF-IL6 knockout mice with wild control mice. Although we examined the cytokine genes expression and induction of such as IL-1 alpha, IL-6, IL-12, IL-18/IGIF, NO2. and TNF alpha in the peritoneal macrophages on 1 month after inoculation, also IL-2 and IL-10 by splenocytes on 1 and 8 months after infection. Following the inoculation of M. leprae with i.p. or BHF, the mice were sacrificed from 1 to 12 months after inoculation in order to confirm the multiplication and the dissemination of the infection. Many leprosy bacilli was found in the peritoneal macrophages of NF-IL6 knockout mice on 1 month after inoculation while that of the wild control mice was showing disappear. In the case of the intraperitoneal infection, NF-IL6 knockout mice shows predominantly multiplication of M. leprae on the abdemino organs such as omentum and also scrotum with male. Although NF-IL6 knockout mice with BHF inoculation did not show any swelling at the site of inoculated foot, however the foot pad on 12 month after inoculation was processed for Fite Faraco's stain and microscopy shows many leprosy bacilli in the intermuscular layer or around the blood vessels/sciatic nerve in the subcutaneous tissue and then the multiplication extended to the toes. Besides the induction of cytokines such as IL-1 alpha, TNF alpha and IL-12 production were observed stronger in culture supernatant of peritoneal macrophages of NF-IL6 knockout mice than that of the wild control mice. IL-2 production was also observed strong in culture supernatant in splenocytes of NF-IL6 knockout mice while that of IL-10 production never induced at anytime. This is doubtless the results of impairment of bacteria killing by macrophages via NF-IL6 gene dependent mechanism not to antigen specific immune system. PMID- 10481453 TI - Brief survey of leprosy situation in Congo: sero-epidemiologic profile in correlation with some emerging viral infections. AB - The Democratic Republic of Congo (DRC, former Zaire) in central Africa remains the foremost country for leprosy in Africa, with a total of 4877 registered cases, of which 4573 are new cases since 1997. These numbers are well above the regional average. About 94% of these patients are under multidrug therapy (MDT) coverage in the Congo, which ranks 8th in coverage rate among the surrounding nine nations. Available data on anatomo-clinical profile and bacillarity are provided, with reservations on the use of these data drwn due to relatively small sample sizes. The seroprofile of the disease was reviewed with regard to the association of other immunity impairing infections like HBV infection and the recently highly incident retroviral epidemics (HIV-1, HTLV-1, and HTLV-2). The leading role of non-governmental organizations is cited for improving leprosy patient conditions and also for future prospects, where the necessity of coordinated strategies with the government is emphasized. Recommendations for new trends and steps relevant to improving existing and future leprosy control strategies are put into perspective. PMID- 10481454 TI - [Rehabilitation with playing therapy for severe multiple handicap people at National Sanatorium, Matsuoka Hoyo-en]. PMID- 10481455 TI - [In Process Citation] PMID- 10481456 TI - [Assessment of pulmonary epithelial permeability in interstitial lung diseases]. AB - The pulmonary epithelial permeability of 99mTc-DTPA (diethylene triamine penta acetate) was assessed in patients with interstitial lung diseases including radiation pneumonitis, idiopathic interstitial pneumonia/pulmonary fibrosis, sarcoidosis, unclassified interstitial pneumonia, and in healthy subjects. Pulmonary epithelial permeability was estimated by the rate constant (kep) of inhaled 99mTc-DTPA clearance from the lungs. Healthy nonsmokers had a mean kep value of 0.82 +/- 0.26% min, and their kep values were constant irrespective of age or sex. Of healthy smokers, 53% showed an increase in kep. This increase correlated with their cigarette consumption per day, but was reversible after cessation of smoking. The provocative concentration of histamine to decrease FEV 1.0 by more than 20% caused an increase in epithelial permeability. However, its effect on permeability was transient, limited, and not dose-dependent. During lung inflation by continuous external negative pressure or by positive end expiratory pressure, pulmonary 99mTc-DTPA clearance was increased, suggesting changes in epithelial permeability. The patients with diffuse interstitial lung diseases also showed increased permeability compared with healthy nonsmokers. In the patients with pre-existing radiation pneumonitis, the mean kep value obtained from the area with infiltration on chest X-ray films was significantly higher than that from the opposite lung. In the prospective study, 3 of 11 patients developed radiation pneumonitis during the course of radiation therapy. The mean kep value obtained in the 3 patients who developed radiation pneumonitis increased just before onset, and further increased when the disease manifested clinically. We believe that 99mTc-DTPA aerosol inhalation is a sensitive test for the detection of inflammatory changes in the bronchioalveolar epithelium. PMID- 10481457 TI - [Three cases of semi-invasive pulmonary aspergillosis occurring in healthy or diabetic individuals]. AB - We encountered three cases of suspected semi-invasive pulmonary aspergillosis, occurring in healthy or diabetic patients. Radiographic findings disclosed cavity formation in 2 of the patients, but only nodular lesions in 1. The beta-D glucan level was elevated in 1 patient, but was within the normal range in the other 2. Morphological findings indicated that all 3 patients had pulmonary aspergillosis. Following the oral administration of FLCZ to 1 patient and ITCZ to the other 2 for about 2 months, the radiographic lesions resolved and hematologic inflammatory reactions cleared up. Gefter et al and Sider et al have defined 2 types of aspergillosis that occur in mildly immuno-compromised or healthy individuals: the semi-invasive type, which destroys pulmonary tissue without vascular invasion and leads to cavity formation; and the locally invasive type, which is characterized by nodular or massive radiographic lesions without cavity formation. Semi-invasive aspergillosis was diagnosed in all 3 of the cases we reported. PMID- 10481458 TI - [Adhesion therapy for malignant pleural effusion (intrapleural administration of OK-432 with minocycline)]. AB - Eleven patients with massive effusion due to pleuritis carcinomatosa were treated by tube drainage, followed by instillation of OK-432 and minocycline for pleurodesis. Pleural immunological and chemical reactions of adhesion were strongly induced by the use of these adhesive agents. As a result, pleural effusion was diminished in all patients without recurrence, allowing the drainage tubes to be successfully removed. As severe adverse effects following this course of therapy, high fever was observed in all patients, and acute renal failure in one. Blood chemical data from the patients revealed an increase in the number of granulocytes with a high level of interleukin-6 one day after instillation. These findings suggested that the symptoms of general inflammation were induced by local pleural inflammation. The median survival period was 253.7 days for 5 patients who were sufficiently fit to be discharged from our hospital. This was better than the historical average for the patients with uncontrolled pleural effusion. In conclusion, it was possible to control malignant pleural effusion and achieve longer survival periods through the optimal management of tube drainage and instillation of adhesion-inducing agents. PMID- 10481459 TI - [Pulmonary tuberculosis presenting multiple nodular shadows in 4 patients]. AB - Four cases of multiple nodular pulmonary tuberculosis simulating metastatic lung cancer were reported. These patients were asymptomatic, found by chance during physical checkups, and had no clinical findings indicative of inflammatory reactions. The nodules ranged from 15 to 50 in number, were spherical in shape, and 3-10 mm in diameter with sharp margins. In 2 patients, the size of the Mantoux reaction (using purified protein derivative) was 22 mm and 11 mm respectively; the other 2 patients were not given the skin test. The final diagnosis was made by video-assisted thoracoscopic surgery in 2 patients, percutaneous lung biopsy in 1, and open lung biopsy in 1. Epithelioid cell granulomas were found in all 4. Anti-tuberculous chemotherapy reduced the size of the nodules significantly in 2 patients and slightly in 1. One patient did not undergo chemotherapy. Although no acid-fast bacilli were detected, all 4 patients were given a diagnosis of pulmonary tuberculosis on the basis of the above findings. Whenever multiple nodular shadows are observed, it is common to suspect metastatic lung cancer at the outset. However, our experience indicated that pulmonary tuberculosis also should be kept in mind as a candidate for differential diagnosis. Computed tomographic findings suggested that the lesions were caused by hematogenous dissemination. Our report therefore devoted some attention to mechanisms of formation. PMID- 10481460 TI - [Eosinophilia and cough induced by resumption of cigarette smoking in a beginning smoker recovering from acute respiratory failure]. AB - A 23-year-old man was admitted to the hospital because of acute respiratory failure accompanied by high fever and severe cough. He had started smoking about 3 weeks earlier and had gradually increased the number of cigarettes he smoked each day. After the number of cigarettes reached 10 per day, the patient experienced cough with serous sputum, high fever (38 degrees C) and dyspnea. Chest X-ray films revealed diffuse, peripheral shadows predominantly in the right lung. The patient was treated with an intravenous drip infusion of antibiotics (FMOX). On the second day of hospitalization, the dyspnea and chest X-ray shadows rapidly resolved. Although the cause of respiratory failure and diffuse shadows on chest X-ray films remained unclear, the patient was discharged. However, on the day of discharge, he smoked cigarettes and experienced cough with serous sputum for a few hours the following morning. He was readmitted for evaluation. We previously reported a case of smoking-induced acute eosinophilic pneumonia (AEP). We suspected that the present case was AEP induced by cigarette smoking but did not perform bronchoalveolar lavage or transbronchial lung biopsy for a definitive diagnosis. Although we did perform a provocation test, the patient demonstrated no respiratory symptoms without cough and respiratory failure. Blood gas analysis and pulmonary function tests also revealed no abnormalities. However, the eosinophil count (3066/mu/l) and percentage (42%) in peripheral blood increased after the patient resumed cigarette smoking, and returned to normal ranges after smoking cessation. These results pointed to a correlation between cigarette smoking and eosinophilia. This case suggested that in some patients, AEP may be induced by cigarette smoking, but does not recur after the resumption of smoking. PMID- 10481461 TI - [Pronounced progression of interstitial pneumonia in a precious metal worker]. AB - A 43-year-old man who had been engaged in the precious-metal processing industry for 18 years was admitted to our hospital because of shortness of breath on exertion. Chest roentgenograms disclosed ground glass opacities and annular nodular shadows in the basal area of the right lung field and almost all of the left lung field. A video-assisted thoracoscopic lung biopsy was performed, and histologic examination disclosed usual interstitial pneumonia (UIP). Mineral analysis revealed high modes of silicon and aluminum, two elements consistent with the materials used in the patient's factory. The histopathological findings indicated interstitial pneumonia caused by the inhalation of dust particles. After corticosteroid therapy, computed tomographic scans disclosed that the ground glass opacities had subsided, but that honeycomb shadows remained. A chest roentgenogram taken 2 years earlier demonstrated only minute granular lesions in the basal area of right lung field. The interstitial shadows had progressed significantly over the intervening 2-year span. The findings in this case should be of value to the etiologic study of UIP. PMID- 10481462 TI - [Hypereosinophilic syndrome associated with antineutrophil cytoplasmic antibody]. AB - A 68-year-old man was admitted to our hospital with complaints of fever, cough, and shortness of breath. He had several erythematous maculae on the trunk and experienced hypesthesia in his lower extremities. Laboratory data showed marked eosinophilia (20,235/mm3) and enhanced hepatobiliary enzymes. Chest X-ray films and computed tomographic scans revealed diffuse patchy infiltrative changes in the lungs. Histologic findings confirmed eosinophilic infiltration of the skin, liver, and lungs. A diagnosis of hypereosinophilic syndrome (HES) was made in accordance with clinical criteria proposed by Chusid et al. The patient was positive for antineutrophil cytoplasmic antibodies (a marker for vasculitis). This suggested a clinical picture resembling Churg-Strauss syndrome (CSS) despite the lack of bronchial asthma. The findings in this report could contribute to a better understanding of the diversity of HES cases, several of which are considered to represent a continuum of pathologies sharing an etiology similar to that of CSS. PMID- 10481463 TI - [Onset of acute eosinophilic pneumonia after resumption of smoking]. AB - We report a case of acute eosinophilic pneumonia (AEP) apparently caused by the resumption of smoking. A 38-year-old woman was admitted to our hospital because of dry cough, high fever and chest pain. Arterial blood gas analysis revealed pronounced hypoxemia and chest X-ray film disclosed diffuse bilateral infiltrate throughout all lung fields. Eosinophils (43.7%) were significantly increased in bronchoalveolar lavage fluid (BALF). The patient was treated with pulse therapy consisting of methylprednisolone and improved quickly. AEP was diagnosed on the basis of BALF findings and the patient's clinical course. Prior to and after her illness, the only noteworthy change in the patient's environmental setting or habits was the resumption of smoking, 3 days after which the symptoms had appeared. We reasoned that the onset of AEP in this patient was strongly related to the resumption of smoking. PMID- 10481464 TI - [Lymphoepithelioma-like carcinoma of the lung]. AB - Lymphoepithelioma-like carcinoma of the lung was diagnosed in a 39-year-old Chinese woman. In situ hybridization of Epstein Barr virus-encoded small nuclear RNA 1 (EBER 1) detected strong EBER 1 signals in the nuclei of tumor cell specimens from the patient. After polymerase chain reaction (PCR) amplification, electrophoresis identified the IR-1 region in this tumor as a positive sharp band, closely resembling Raji cells (Burkitt's cell line). The uniform and intense presence of EBER 1 in the tumor nuclei and the PCR products of EBV DNA in the tumor demonstrated that a high copy number of EB virus genome existed in the tumor, and indicated involvement of the EB virus in the pathogenesis of lymphoepithelioma-like carcinoma of the lung. PMID- 10481465 TI - [Sarcoidosis with hypercalcemia, urolithiasis, renal insufficiency, and heart failure]. AB - A 65-year-old man was admitted to our hospital because of thirstyness and left lower abdominal pain. On admission, he was found to have urolithiasis, renal insufficiency (BUN: 73 mg/dl, Crt: 4.4 mg/dl), and hypercalcemia (13.2 mg/dl). Chest X-ray films and computed tomograms showed enlargement and calcification of the hilar lymph nodes, and thickened interlobar fissures in both lungs. Levels of angiotensin converting enzyme (30.2 IU/l) and 1.25 (OH)2VitD3 (66.4 pg/ml) were elevated. Histologic examination of the specimen obtained from transbronchial lung biopsy showed non-caseous epithelioid cell granulomas. Because the level of parathyroid hormone was normal and no malignancies were detected, a diagnosis of sarcoidosis was made. Treatment with extracorporeal shock wave lithotripsy, transurethral lithotomy, saline infusion, and prednisolone (30 mg/day) alleviated the urolithiasis, renal insufficiency, and hypercalcemia. After discharge, the patient was followed up and given prednisolone therapy. About 1 month after the prednisolone dose had been tapered to 15 mg/day, the patient experienced dyspnea and facial and pedal edema. Because congestive heart failure was diagnosed, he was re-admitted to our hospital for a second time. Although he was then placed on intensive therapy, he died of ventricular tachycardia associated with sarcoidosis of the heart. PMID- 10481466 TI - [Lung cancer with small intestine metastasis characterized by exceptionally high levels of serum CA 19-9]. AB - A 59-year-old man was given a diagnosis of lung cancer (moderately differentiated tubular adenocarcinoma) with left adrenal gland and bone metastases in January 1997, and received chemotherapy and irradiation therapy. In late May, anemia and occult blood were detected, with a marked increase in serum CA 19-9. In August, the patient was admitted to our department complaining of melena. His serum CA 19 9 level on admission was significantly elevated (18,960 U/ml). After admission, symptoms of ileus developed. Radiographs of the small intestine and abdominal computed tomographic scans suggested the presence of a tumor in the small intestine. Therefore, surgery was performed, revealing a tumor in the jejunum, which was histologically diagnosed as metastasis of lung cancer to the small intestine. Immunohistochemical staining for CA 19-9 was more intense in specimens from the small intestine tumor than from lung cancer specimens. Serum CA 19-9 decreased significantly after resection. The clinical course and results of CA 19 9 staining suggested that CA 19-9 production by the metastatic lesion in the small intestine was the major cause of the patient's high serum CA 19-9 level. This appears to be a rare case because, to our knowledge, there are no previous reports in the Japanese literature on patients with small intestine metastasis from lung cancer showing an exceptionally high level of serum CA 19-9. PMID- 10481467 TI - [Exogenous lipoid pneumonia induced by inhalation of insecticide]. AB - We report a case of exogenous lipoid pneumonia induced by insecticide. A 67-year old man was admitted to our hospital with complaints of dyspnea cough, hemoptysis, and shivering 6 hours after inhaling an insecticide. A chest radiograph on admission disclosed consolidation in the right upper lobe. Chest computed tomography showed nodular infiltrates associated with ground-glass opacities. Bronchoalveolar lavage fluid was hemorrhagic and showed neutrophilia. Pathological examination of transbronchial lung biopsy specimens showed numerous vacuolated macrophages in alveolar spaces. Electron microscopy demonstrated lipid droplets in the cytoplasm of these macrophages. These findings together yielded a diagnosis of exogenous lipoid pneumonia induced by insecticide. In addition, we experimentally induced a similar form of lipoid pneumonia in rats by intratracheal instillation of the same insecticide. PMID- 10481468 TI - [Lung and bone marrow granulomas associated with human parvovirus B19 infection]. AB - A 56-year-old man was admitted with flu-like symptoms, and his platelet count abruptly decreased. A chest X-ray film showed granular shadows, and lung and bone marrow specimens disclosed non-caseating epithelioid cell granulomas. The patient's serum IgM titer for human parvovirus (HPV) B19 was elevated. Our conclusion was that HPV B19 must be kept in mind as a possible pathogenic agent of granuloma formation. PMID- 10481469 TI - [Chronic pulmonary thromboembolism diagnosed on the basis of characteristic mosaic patterns on lung computed tomograms]. AB - We report on a 59-year-old woman who presented with characteristic findings on lung computed tomographic (CT) scans and was therefore suspected to have chronic pulmonary thromboembolism. She visited our hospital because of worsening exertional dyspnea over the preceding year, and because she was dissatisfied with an earlier diagnosis made by another institution. Chest roentgenograms disclosed nonhomogeneous hyperlucency in both lungs associated with linear and bundle shadows, dullness of the right costophrenic angle, and dilatation of the descending branch of the right pulmonary artery. The patient experienced moderate hypoxemia even at rest. Pulmonary function tests demonstrated a restrictive ventilatory pattern associated with reduced diffusion capacity. The lung CT scans disclosed a mosaic pattern of attenuation in the lung parenchyma, which probably reflected scattered areas of low perfusion. The unique characteristics of such CT findings drew our attention to chronic pulmonary thromboembolism as a possible diagnosis. We eventually confirmed the diagnosis on the basis of enhanced CT scans, pulmonary perfusion and ventilation scintigrams, and digital subtraction angiography. In our view, chronic pulmonary thromboembolism should be kept in mind as a possible differential diagnosis of the mosaic patterns of attenuation on lung CT scans. PMID- 10481470 TI - [Function and nerve growth factor of the rat urinary bladder during urethral obstruction and its relief]. AB - It has been considered that the urethral obstruction influence the function and the nerve innervation of the bladder. In this study, the changes in the bladder function and the nerve growth factor (NGF) synthesized in the bladder were systematically analyzed both during partial urethral obstruction and after its relief in the rat. The maximum contraction pressure was temporally decreased and then increased 1.5 times of the normal level 6 week after the obstruction. NGF in the bladder, measured by ELISA method, was rapidly increased 5 times of the normal level 1 day after the obstruction. It gradually decreased 2 weeks after the obstruction but did not recover to the normal level. The maximum contraction pressure and the bladder NGF recovered to the normal level after the removal of 1 to 6 weeks obstruction. These results suggest that partial urethral obstruction reversibly increases the contractility and the NGF synthesis of the bladder. PMID- 10481471 TI - [Examination for indication of systematic biopsy for diagnosis of prostate cancer]. AB - BACKGROUND: Systematic biopsy has been commonly used for detection of prostate cancer. Nevertheless, as this examination occasionally gives patients severe complications it is necessary to give careful consideration for application of this examination. Thus, we analyzed retrospectively 145 cases who underwent transrectal ultrasonography (TRUS) guided systematic biopsy to evaluate the application of systematic biopsy, correlating with the findings of digital rectal examination (DRE), prostate specific antigen (PSA), the findings of transrectal ultrasonography (TRUS) and the results of biopsies. METHODS: Between May, 1995 and May, 1997, 143 patients who were suspected to have prostate cancer with either of PSA and DRE, and 2 patients who received visual laser ablation of prostate (VLAP), underwent TRUS guided systematic biopsy of prostate. We evaluated diagnostic efficacy of PSA, DRE, TRUS, prostate-volume-specific PSA, and PSA density (PSAD). RESULTS: Sensitivity, specificity and positive predictive value (P.P.V.) are 78.4%, 62.8% and 53.5% for DRE, 100.0%, 4.4% and 41.8% for PSA, 88.2%, 60.0% and 52.9% for TRUS, 87.8%, 72.1% and 64.2% for prostate-volume specific PSA, 100.0%, 30.6% and 45.4% for PSAD, respectively. Ten of 69 patients (14.5%) whose PSA levels were 4.0 to 10.0 ng/ml were diagnosed as cancer, and positive for both or either of DRE and TRUS. Twenty-seven who were negative for both of DRE and TRUS were not diagnosed as prostate cancer. Using the combination of prostate-volume-specific PSA, DRE and TRUS, we could eliminate 29 non-cancer men (21.5%) whose PSA level was greater than 4.0 ng/ml from systematic biopsy. CONCLUSION: On the diagnosis of prostate cancer, the combination of prostate volume-specific PSA, DRE and TRUS is very useful to exclude unnecessary systematic biopsy, if an urologist could be used to and trained for DRE and TRUS. PMID- 10481472 TI - [Clinical review of acute scrotum in children]. AB - BACKGROUND: The acute scrotum in children frequently presents a diagnostic dilemma. The objective of this study is to review the experience of patients with acute scrotum in children. METHODS: From April 1986 until March 1998, we encountered 40 cases of acute scrotum in children less than 15 years old diagnosed by surgical exploration. RESULTS: Final diagnoses consisted of 14 patients with torsion of the spermatic cord (35%), 22 patients with torsion of the testicular appendage (55%), 3 patients with acute epididymitis (7.5%), and patient with a strangulated hernia (2.5%). Of 22 patients with torsion of the testicular appendage, which were the most frequent cause of acute scrotum, 20 had torsion of the testicular appendix and 2 had torsion of the epididymal appendix. The age distribution of the patients with torsion of the spermatic cord was biphasic, with the highest frequencies occurring in patients 2 years old and 14 years old. In contrast, the age distribution of patients with torsion of appendage showed a single peak incidence at 9 years of age. Those with epididymitis were younger than 2 years old. Of 14 cases of torsion of the spermatic cord, orchiectomy was performed in 6 cases (42.9%) because of testicular necrosis. CONCLUSION: Although surgical exploration is no longer necessary for all patients with torsion of the appendage, this review revealed that patients complaining of severe or persistent pain usually require surgery to diagnose and treat the problem. PMID- 10481473 TI - [Clinical study of grade 3, superficial transitional cell carcinoma of the urinary bladder]. AB - PURPOSE: To evaluate the treatment of grade 3 superficial (stage pTa and pT 1) transitional cell carcinoma (T.C.C.) of the urinary bladder, retrospective analysis was performed with special reference to tumor prognostic factors. MATERIALS AND METHODS: From July 1971 to september 1995, 51 cases with grade 3 superficial T.C.C. of the urinary bladder were treated. The survival rates and prognostic factors of these patient were analyzed. RESULTS: Five year survival rate of grade 3, superficial tumors was 92.3% and showed significantly better prognosis compared to patients with pT 2 and pT 3 tumors of grade 3 (p < 0.001). As a initial treatments, transurethral resection (TUR) was conducted in 45 patients (88%). Intravesical recurrence was observed in 20 of 45 patients (44%) and 12 of 20 patients (60%) were recurred within 1 year. Non-recurrent rates of the patients treated with TUR were 69.6% at 1 year, 58.8% at 3 year, 49.7% at 5 year, respectively. No significant differences were noted regarding factors of tumor size, figures and a number of tumor. Of the 51 patients, 10 (19.6%) progressed beyond stage T 2 and 6 died with the disease. Survival rates at 10 years follow up in patients with non-papillary and papillary tumor were 57.1% and 97.8%, respectively. CONCLUSION: These results suggested that TUR should be performed as a initial treatment for the patients with grade 3 superficial T.C.C. of the urinary bladder. However, non-papillary tumors should be considered of more intensive treatment like as radical cystectomy, adjuvant chemotherapy or irradiation. PMID- 10481474 TI - [The evaluation of methods for determinations of urinary oxalate]. AB - BACKGROUND: Estimation of urinary oxalate is one of important tools for the diagnosis and treatment of urolithiasis. And more precise, simple and inexpensive method is desirable. In the present study, we evaluated three methods which were clinically well-used. METHODS: From October 1996 to June 1997, 146 acidified urine samples were collected for 24 hours from 144 urolithiasis patients. We determined the urinary oxalates by three methods; the colorimetric method, the enzymic method and the ion chromatography (IC method). And we evaluated the correlations of these methods. RESULTS: Correlation coefficients in the urine oxalate concentration were, 0.86 with the colorimetric method and the IC method, 0.91 with the enzymic method and the IC method, 0.90 with the colorimetric method and the enzymic method. The coefficients in the 24 hours-urinary excretion of oxalate (0.76, 0.87, 0.82) were lower than those in the urine oxalate concentration. The correlation coefficients with the colorimetric method and the IC method were, 0.58 in hyperoxaluric group, 0.34 in normooxaluric group. The coefficients with the enzymic method and the IC method, 0.93 in hyperoxaluric group, 0.71 in normooxaluric group. CONCLUSION: The colorimetric method is least expensive, but is less useful. The enzymic method is less expensive, and is as useful as the IC method. PMID- 10481475 TI - [The role of ATP-receptor in controlling the urinary bladder and urethral function in rats]. AB - BACKGROUND: The detrusor contraction involves an atropine resistant, nonadrenergic noncholinergic (NANC), component. Since adenosine triphosphate (ATP) has been proposed as a NANC transmitter, the role of ATP-receptors in the lower urinary tract was examined. MATERIALS AND METHODS: The isovolumetric bladder contractions and the urethral pressure were monitored after intra arterial administration of ATP analogues and other drugs in 52 female SD rats under intraperitoneal urethane anesthesia. RESULTS: alpha beta metATP induced a rapid, phasic increase of the maximal bladder pressure immediately after the drug administration, which was followed by a decrease during the P 2 X-purinoceptor desensitization period, but it did not affect the resting bladder pressure. RB-2 did not affect both the maximal bladder pressure and the resting bladder pressure. As for the urethral functions, alpha beta metATP induced a decrease of the resting urethral pressure during the P 2 X-purinoceptors desensitization period, while RB-2 induced a increase of the resting urethral pressure in contrast. ATP, alpha beta metATP and RB-2 did not changed the maximal urethral relaxation. CONCLUSION: The results will indicate that ATP produces detrusor contractions through the P 2 X-purinoceptor. Although ATP dose not affect urethral relaxation during voiding phase, it controls the urethral tone during collecting phase by both the excitation of P 2 X-purinoceptor and the inhibition of P 2 Y-purinoceptor. PMID- 10481476 TI - [Extravasation into a renal sinus cyst due to ureteral calculus: a case report]. AB - A 71-year-old man who had been treated conservatively for advanced prostate cancer with hormonal therapy presented with complaint of left flank pain. Intravenous urography demonstrated obstruction caused by a left upper ureteral calculus and leakage of contrast medium outside the renal pelvis and calyces. Left percutaneous nephrostomy was performed, and subsequent nephrostography showed extravasation into a parapelvic renal sinus cyst previously recognized on follow-up computed tomography (CT). The left ureteral calculus was treated with extracorporeal shock wave lithotripsy (ESWL), after which obstruction and extravasation disappeared. One month later, abdominal CT demonstrated no left renal nor ureteral calculi, as well as a reduction in the left renal sinus cyst. Urinary extravasation sometimes occurs as a complication of acute urinary obstruction. However, extravasation into a cyst is very rare. We have found only one case reported in the literature, so far. PMID- 10481480 TI - [NO and pancreatic diseases]. PMID- 10481477 TI - [Bilateral synchronous multilocular epididymal cysts: a case report]. AB - A rare case of bilateral synchronous multilocular epididymal cysts is reported. A fifty-six year old man visited to our hospital with a chief complaint of swelling of bilateral intrascrotal contents. Ultrasonographic findings demonstrated multilocular lesions of the bilateral intrascrotal contents. Preoperative diagnosis was bilateral multilocular hydrocele testes. Operative procedure revealed bilateral cysts originating from the head, body and tail of the epididymis without the cysts of the tunica vaginalis. Operative sight was bilateral synchronous multilocular epididymal cysts, and bilateral epididymal cystectomy were performed. The specimen size was right diameter 12 x 6 cm and left its 8 x 5.5 cm. The puncture of the cystic fluid revealed many spermatozoa in both sides. The acquired cysts of the epididymis generally are the result of tubular obstruction with dilation of tubules adjacent to the obstruction. The dilated tubules are filled variably with viable and degenerating spermatozoa. The cysts originate most commonly from the head of the epididymis, and are unilateral, unilocular or multilocular and are usually within 1 cm in diameter. In our case, bilateral synchronous epididymal cysts originating from not only the head, but also the body and the tail is a rare case. PMID- 10481482 TI - [Clinical study on liver metastasis of AFP-producing gastric cancer- characteristic findings of angiography]. AB - We studied 6 patients with liver metastasis of AFP-producing gastric cancer, especially about the angiography and treatment. The angiographic findings of liver metastasis are characteristic in 3 patients. Multiple rounded hypervascular tumor are seen in the arterial phase and its vessels are fine. In the venous phase, the tumor is homogeneous. We called the findings "Fireworks like stain". These patients were treated with TAE and responded it. PMID- 10481481 TI - [The expression of chemokines and the dynamics of inflammatory cell infiltration before and after H. pylori eradication]. AB - We attempted to evaluate the relationship between the expression of IL-8 and RANTES and the dynamics of their target cells in human gastric mucosa of H. pylori associated gastritis, including their changes after H. pylori eradication. We performed the measurement of the mucosal level of IL-8 and RANTES protein by ELISA and immunohistochemistry. The neutrophil infiltration into the gastric mucosa was identified by the histological examination based on the Updated Sydney system and the measurement of MPO activity. The memory T lymphocyte and eosinophil were indicated by immunohistochemistry of CD45RO that is one of surface markers indicating memory T lymphocytes and MBP that is contained in the granules of eosinophils. H. pylori positive gastric mucosa demonstrated a remarkable increase in neutrophils. CD45RO positive cells and eosinophils, compared to H. pylori negative gastric mucosa. Gastric mucosal level of IL-8 and RANTES protein and MPO activity was significantly higher in H. pylori positive cases than that in H. pylori negative controls after H. pylori eradication, both of the level of IL-8 protein and MPO activity reduced at the same levels as negative controls. However, RANTES expression, CD45RO positive T lymphocytes and eosinophils remained in H. pylori eradicated gastric mucosa at the significantly high level, compared with H. pylori negative cases. Therefore, it seems possible that IL-8 might enhance the inflammation by facilitating the neutrophil infiltration into H. pylori infected gastric mucosa and that RANTES might play an important role in the specific immune response against H. pylori and the maintenance of the immune memory after H. pylori eradication. PMID- 10481483 TI - [Thrombotic thrombocytopenic purpura associated with advanced gastric cancer]. PMID- 10481484 TI - [A case of triple cancers with gastric, jejunal and colonic cancers]. PMID- 10481485 TI - [A case of lymphangioma of the small intestine]. PMID- 10481486 TI - [The cases of Fitz-Hugh-Curtis syndrome]. PMID- 10481487 TI - [A case of ileal diverticulum with a foreign body manifested by digestive tract bleeding]. PMID- 10481488 TI - [A case of duct-islet cell carcinoma of the pancreas positive for somatostatin immunostain]. PMID- 10481489 TI - [A case of calcified false cyst of the spleen]. PMID- 10481492 TI - [Hidden chromosome instability and risk of laryngeal cancer incidence]. AB - The bleomycin test is a recognised method of evaluation of hidden genetical instability. The concept of the test consists in inducing chromosome aberrations in lymphocytes exposed in vitro to bleomycin and subsequent quantitative analysis of chromosome breaks. The study material was whole venous blood from 61 laryngeal cancer patients and from 30 healthy persons taken as a control. For each patient two parallel cultures where carried out in the standard procedure. Bleomycin was added to one of the cultures to induce chromosome breaks. Then, in microscopic metaphasal plates stained by Giemsa dye, the chromosome instability index estimated as a number of chromosome breaks per cell (b/c) and the percentage of cells with chromosome breaks were calculated. Higher indices of chromosome instability were demonstrated in laryngeal cancer patients in comparison to the controls. The persons with chromosome instability (b/c > 0.8) or with chromosome oversensitivity to mutagens (b/c > 1) were identified only among larynx cancer subjects. Furthermore, it was established that an increased chromosome instability is associated with high aggressiveness recognised by histological grading. The latter finding requires confirmation on an enlarged group of subjects. PMID- 10481493 TI - [Estimation of bleomycin-induced chromosome aberrations in lymphocytes of laryngeal cancer subjects. Preliminary report]. AB - Chromosome instability is associated with an increased risk of malignancy. However, the quantitative analysis of chromosome breaks provided by the bleomycin test requires additional analysis aimed for the localisation of chromosome aberrations. For this reason, the metaphasis slides prepared for bleomycin test were stained with fluorochrome DAPI to estimate chromosome breaks in particular chromosomes. The additional staining of chromosomes can be recognised as an extension of the classical bleomycin test addressed for identification of structural aberrations. Preliminary results indicate that the most frequent chromosome breaks were found in chromosomes 1, 2, 3, 7 and 13. PMID- 10481494 TI - [Compensatory mechanisms of food transmission and lower respiratory tract protection after classical and extended supraglottic laryngectomies. Computerized topokinetic analysis of roentgen-cinematographic images]. AB - Studies were performed on 22 patients, aged from 17 to 78 years, in whom, owing to laryngeal cancers, partly classical or extended supraglottic laryngectomy was carried out. The evaluation involving the pharyngeal deglutition course was accomplished by resorting to computerized topokinetic analysis of the roentgen cinematographic images. The completed observation revealed good mobility of the anatomical structures participating in deglutition, small volume and number of glossolaryngeal recesses. There were few patients in whom the contrast medium passed to the trachea. Better passage of food as well as protection of lower respiratory tract were associated with the improvement of parameters facilitating the deglutition. PMID- 10481495 TI - [The content of some trace elements in tissue of laryngeal squamous cell carcinoma]. AB - The examination of the content of arsenic, nickel, copper, selenium, zinc, iron, cadmium, lead, cobalt, antimony and tin in the tissue of laryngeal squamous cell carcinoma was carried out. Material was taken from tumour of the larynx of 22 patients during laryngectomy. The control group was made up by healthy tissue of the larynx taken from 14 patients during diagnostic examinations. Trace elements were determined by means of BAIRD plasma spectrometer. Higher values of the concentration of arsenic, nickel, copper, cadmium, lead and cobalt were found in the tissue with carcinoma of the larynx as compared with the control group. The concentration of selenium, zinc, and iron in the carcinoma tissue was lower than in the healthy tissue. The concentration of antimony and tin was similar to that of the control group. PMID- 10481496 TI - [Lymphorrhea after neck dissection]. AB - In this publication lymphorrhea was described as one of possible complications after the neck dissection surgery of Crile-Jawdynski procedurae. The matter of this complication is rise of pressure in lymph system. 4 women with this complication after neck dissection, the best treatment is drainage of the operated area. In one patient, because of the failure of conservative treatment, another surgical intervention was preformed. The full treatment of lymphorrhea was after 4-6 weeks. PMID- 10481497 TI - [Primary neoplasms of the parapharyngeal space in material from the Otolaryngology Department, Karol Marcinkowski University of Medical Sciences in Poznan]. AB - Primary neoplasms of parapharyngeal space are rare. In this location the so called non-malignant tumors occur many times more than the malignant ones. In our paper, employing statistics, objective symptoms and complaints connected with these tumors were ordered according to frequency. In recent five years in ENT Department of Karol Marcinkowski University of Medical Sciences in Poznan 9 persons were surgically treated. The differentiated histopathological picture is discussed and described (6 malignant and 3 non-malignant). It can be claimed that MR is more useful in diagnosis than CT. The treatment is usually surgical; both external and oral approaches are used. PMID- 10481498 TI - [Foreign bodies in the esophagus]. AB - 1496 patients with foreign bodies of esophagus, reported between 1945-1997 to ENT Department of K. Marcinkowski University of Medical Sciences, were analysed. The age ranged from 0.5 to 95 years, but the most numerous group was under 5 years of age. Men outnumber women. Very detailed anamnesis, oropharynx and hypopharynx examination, neck palpation, chest X-ray were crucial for diagnosis. More than 50% of patients reported within first 10 hours after foreign body wedged. The most often met foreign bodies were: bones, coins, denture, fish bones, fruit stones, buttons. The most dangerous were supposed to be: needles, pieces of glass, springing anchores, safety razors. Foreign bodies were removed by means of rigid esophagoscopes. In 75.3% foreign bodies occupied the first isthmus. Serious complications met in the analysed material, included: esophageal wall perforation [27], mediastinitis [21], aortal hemorrhage [3], esophagus-tracheal fistula [1]. Intramural abscesses, posteriol pharyngeal wall abscesses were found more often. 98% of esophageal foreign bodies were removed by the same route they had been introduced. PMID- 10481499 TI - [Hormonal function of the thyroid gland after therapy for laryngeal cancer]. AB - The paper presents the results of the study concerning thyroid gland function of the patients suffering from larynx cancer treated with: a) surgery only, b) surgery with further radiotherapy with radical doses 5000-6500 cGy, c) radiotherapy along with radical doses 6500-7200 cGy. Thyroid gland function was evaluated by measurement of: a) hTSH, FT3, FT4, b) serum lipids concentration. The hTSH concentration was evaluated by immunoradiometric method whereas FT3 and FT4 by DELFIA techniques (deleted fluorescence). 162 patients (M-148, F-14) were examined. Average age was 57 (min.-30, max.-80). The patients were divided into two groups. The first group (n = 90) involved those who had been examined once at different time since the end of treatment (retrospective examinations). The patients who had been examined before as well as after treatment were established as the second group (n = 72) (prospective examinations). In this case the examinations are still being continued and they are carried out at: a) 1, b) 1-6, c) 6-12, d) 12-24, e) longer than 24 months after completion of treatment. Our preliminary data show that hypothyroidism (mostly subclinical) occurs seldom (7% of all the cases) in the early phase (up to 1 month) after surgical treatment and gets normal spontaneously (up to 6 months). After radiotherapy only--no subclinical hypothyroidism was observed. Interestingly, the combined treatment seems to be the most frequent cause of subclinical hypothyroidism. Its frequency increases (up to 47%) with the length of time since the conclusion of treatment (2 years and longer). Because of continuation of our examinations the presented data are still preliminary and they will be verified in the next paper. We also tried to explain the pathogenesis of subclinical hypothyroidism on the basis of histopathologic examination of thyroid tissue derived from the patients (after combined treatment and after radiotherapy) during further total laryngectomy performed because of neoplasm renewal. PMID- 10481500 TI - [Evaluation of selected cellular immunity parameters in patients with seasonal allergic rhinitis]. AB - The aim of the study was the estimation of selected parameters of cellular immunity in patients with seasonal allergic rhinitis and also the estimation of histamine effect on T lymphocytes. The study comprised 240 persons--180 patients with seasonal allergic rhinitis, 30 patients with nonallergic rhinitis (positive control group) and 30 healthy persons (negative control group). In all persons the following examinations were carried on: allergic skin tests, serum level of IgE, Multitest CMI, percentage of lymphocytes T (CD3), percentage of lymphocytes T helper (CD4), percentage of lymphocytes T cytotoxic (CD8) and influence of histamine on T lymphocytes in vitro. The analysis of acquired results suggests that the inflammatory changes in patients with seasonal allergic rhinitis arose from hypofunction of T lymphocytes, especially of lymphocytes T cytotoxic (CD8) in their reaction with histamine. PMID- 10481501 TI - [Evaluation of internal nose deformation in patients with unilateral cleft lip and palate]. AB - The methods of assessment of nose deformation in patients with unilateral cleft lip and palate have been presented. The study covered 45 patients with clefts, of 14-20 years of age who had undergone the infant surgery, and 45 patients at the same age serving as controls. The condition of nasal septum has been clinically examined in both groups and the nasal resistance has been measured by means of front and rear rhinomanometry. In the cleft group 80% of patients presented their cartilaginous septum deviated to the non-cleft side, and 85% had their bony septum deviated to the cleft side. In the cleft group, the front rhinomanometry revealed significantly higher resistance on the cleft side than on the non-cleft side and in controls. The rear rhinomanometry showed no significant differences in both groups, except the patients with clefts, who had been submitted to surgery with pharyngeal flap, in whom higher values of resistance had been found. PMID- 10481502 TI - [Otoacoustic emission nonlinear distortions in musicians with absolute and relative pitch]. AB - In the presented study an analysis was performed of the properties of the cochlea using Distortion Product Otoacoustic Emissions (DPOAE). 52 musicians were examined, aged 17-31 years, with systematic musical training longer than 10 years, and 30 non-musicians without musical experience, aged 18-30 years. Then all musicians were tested with Rakowski's test to divide them into the absolute pitch possessors (APP) and relative pitch possessors (RPP). DPOAE recorded in musicians were statistically higher than in nonmusicians for f2 = 1.0, 2.0 and 4.0 kHz for signal intensities generally ranged from 45 to 55 dB SPL. Analogous comparison between APP and RPP showed higher DPOAE for f2 = 2.0 and 4.0 kHz in APP but the differences were not statistically significant. Simultaneously no significant differentiations were observed between investigated groups for highest signal intensities in the whole frequency band. The study suggests that systematic musical training influences the cochlear mechanics assessed by DPOAE and the greatest differences concern the active processes within the cochlea. The continuation seems to be justified. PMID- 10481503 TI - [Low-power laser in the treatment of tinnitus--a placebo-controlled study]. AB - The present study was performed on 32 patients in order to investigate the effect of low-power laser on their tinnitus. The patients were divided into two groups. One group received laser therapy and the other was given a placebo procedure. The effect was evaluated by the use of visual analogue scales. Within the patient group transiently evoked otoacoustic emissions (TEOAE) were measured before, during and after therapy. No significant difference between laser and placebo was found in annoyance or loudness of the tinnitus and in changes of TEOAE amplitude. These results indicate that there is no relationship between the effect of low power laser and changes in cochlear micromechanics. PMID- 10481504 TI - [Speech disorders in Alajouanine syndrome]. AB - Alajouanine syndrome includes congenital, bilateral, central facial and abducens nerves palsy and clubfoot. The authors present speech disorders and helpful influence of early rehabilitation. PMID- 10481505 TI - [Cases of pleomorphic adenoma originating in the minor salivary glands]. AB - The authors present 6 cases of adenoma pleomorphum localized out of salivary glands-2 of them on palate, 1 in external auditory canal, 1 in larynx, 1 in lacrimal gland and 1 in parapharyngeal space. All patients were treated surgically. PMID- 10481506 TI - [Aneurysmal temporal bone cyst]. AB - Two cases of aneurysmal temporal bone cysts treated surgically are described. PMID- 10481507 TI - [Melkersson-Rosenthal syndrome]. AB - The Melkersson-Rosenthal syndrome consists of a triad of symptoms: peripheral facial nerve paralysis, congenital "lingua plicata" and noninflammatory facial oedema. The authors present 2 cases with complete picture of this syndrome and some review of the literature. PMID- 10481508 TI - [Adenolymphoma (Warthin's tumor) located bilaterally]. AB - Adenolymphoma (Warthin's tumor) is the second in occurrence of the parotid gland's benign neoplasms. In most cases (80%) it is observed as an unilateral tumor, smoking males aged from 50 to 60 are mainly affected. The authors present an interesting case of simultaneously occurring bilateral Warthin's tumor growing in parotid gland. A 59-year-old male patient, smoker was admitted to E.N.T. Dept. with a bilateral tumor. He underwent surgical treatment: superficial parotidectomy on the left side and right side tumor exterpation at the same time. After the operation we got the confirmation of the preoperation diagnosis. The patient was discharged home and stayed on the follow-up. Smoking patients with diagnosed unilateral adenolymphoma should be followed up because of the possibility of the neoplasmatic recurrence. PMID- 10481509 TI - [The use of CO2 laser in otolaryngology]. AB - The paper presents the problems concerning technology and application of laser beam in ENT. The author provides a summary of the current status of this field. This work facilitates the knowledge of physical, technical and medical activities of CO2 laser beam on different human tissues. It also describes the potential applications of CO2 laser technique in the treatment of different ENT conditions. PMID- 10481510 TI - [Decongestants in treatment of nasal obstruction]. AB - Decongestants have long been used in nose obstruction. On the one hand they seem to be abused in local treatment, on the other they are too rarely used in initial treatment of other pathological states of the nose. Yet, as the knowledge of the nasal mucosa vascular system and its neurological and humoral regulation increases, there appear broader indications for their application. A more frequent use of the drugs could be suggested in infectious and allergic rhinitis and paranasal sinusitis, where their application prevents complications and enhances the efficacy of treatment. The sympathomimetic drugs are recommended in rhinitis because a reduction of nasal congestion permits the introduction of other topical therapy such as nasal corticosteroids. They are also recommended in rhinitis during pregnancy and diagnostic procedures in some cases of structural rhinitis. This article briefly discusses the relationships between the nasal mucosa vascular system, the autonomic nervous system and neuropeptides in the regulation of air flow in the nose. Regarding their use in rhinitis treatment, there were characterized groups of sympathomimetic vasoconstrictor agents, which include sympathicomimetics (e.g. phenylpropandamine, phenoloephedrine) and alpha receptor agonist medications (e.g. xylometazoline, oxymetazoline, naphazoline). The author discussed the pathomechanism of rhinitis medicamentosa resulting from abuse of nasal drops containing alpha-agonists and suggested a therapy. PMID- 10481511 TI - [The fiftieth anniversary of Miodonski's electro-rhino-spirometry]. AB - On the 50th anniversary of the introduction of the electro-rhino-spirometry by Miodonski the author recalls its theoretical foundations and diagnostic possibilities. Moreover, it has been stressed that Miodonski's method was the first one which does not disturb the nasal function, and the development of electronics supported and widened its diagnostic possibilities leaving the theoretical basis for examination unchanged. PMID- 10481512 TI - [Otolaryngologic hospital wards and out-patient clinics in Warsaw in the second part of the XIX-th and the beginning part of the XX-th century]. AB - The origin of the first otolaryngological hospital wards in Warsaw: laryngological ward of St. Roch Hospital and otological ward of Ujazdowski Hospital, both founded in 1881, and laryngological out-patient clinic at St. Spirit Hospital, founded in 1883, is described. The rise of the first otolaryngological hospital ward in Warsaw Orthodox Jew Hospital established in 1903 is thoroughly described. The activity of Warsaw hospital wards and hospital out-patient clinics at Infant Jesus Hospital, the Warsaw Hospital for Children, the Berson and Bauman Hospital, the Wolski Hospital, the Blessed Virgin Mary Hospital, the Evangelical Hospital, the Charles and Mary Hospital for Children, the Warsaw Municipal Lazaret is presented in detail. PMID- 10481513 TI - [Selected biochemical tumor markers in clinical monitoring of head and neck neoplasm progression]. PMID- 10481514 TI - [Report from the academic workshop on: "New objective examination methods in audiology and otoneurology". Warsaw, September 24-26, 1998]. PMID- 10481515 TI - [Report from the VII International Pediatric Otolaryngologic Congress in Helsinki. June 7-10, 1998]. PMID- 10481516 TI - [Report of the III International Symposium on the topic: "Experimental Rhinology and Immunology of the Nose". Gent, Belgium, November 12-14, 1998]. PMID- 10481517 TI - [Report and World Congress on Head and Neck Oncology. Madrid, November 29 December 3, 1998]. PMID- 10481518 TI - [Noninvasive mechanical ventilation]. PMID- 10481519 TI - [Function of peripheral airways in children and adolescents with deformities of the anterior chest wall]. AB - Body-plethysmography and spirometric test were carried out in two groups of 33 children aged 7-16. Examined group consisted of 11 children with deformities of rib chest wall, and control group consisted of 22 healthy children. Pulmonary function test were performed in all subjects. They comprised spirometric parameters (FVC, FEV1, ITGV, RV, TLC) flow-volume curve parameters (PEF, FEF25, FEF50, FEF75) and airway resistance R(aw). Values of FEV1, FEF25, FEF50, FEF75 PEF and MMEF were significantly lower in the tested group than in the control group. In children with deformities of rib chest wall values of airway resistance were significantly higher, R(aw)--268%N, than in healthy children, R(aw)--188.5%N (p < 0.01). PMID- 10481520 TI - [Endothelin concentration in sera of patients with atopic bronchial asthma]. AB - Endothelin-1 (ET-1) is an effective vasoconstrictor and has bronchoconstricting property. Recent findings in vivo and in vitro indicate the influence of ET-1 on bronchial smooth muscle tone. ET-1 concentration was detected in BAL-Fluid in patients with bronchial asthma. Previous studies indicated an increase in ET-1 serum concentrations in the course of exacerbation of asthma. The purpose of our study was to compare ET-1 concentrations in sera of asthmatics during the asymptomatic period and after metacholine provocation. The study was performed in a group of 16 patients with mild bronchial asthma and in 11 healthy subjects. ET 1 concentrations in sera were evaluated by Elisa method (kit R&D USA). Bronchial provocation with metacholine was performed in asthmatics using Bronchoscreen. The result was considered as positive with FEV1, decrease of at least 20%. Airway responsiveness to metacholine was expressed as PC20. The results were analyzed statistically by means of the Student's test. The baseline ET-1 levels in sera of healthy donors were lower then in asthmatics group (x1' = 4.72 +/- 1.01, x1 = 11.53 +/- 3.68 pg/ml, p < 0.001). We found a statistically significant increase of ET1-1 concentration in sera after inhalation of metacholine (x1 = 11.55 +/- 3.68; x2 = 24.08 +/- 4.09 pg/ml, p < 0.001). The results indicate that ET-1 may play a role in bronchospasm in asthmatics. PMID- 10481521 TI - [Comparison of exercise and histamine provocation tests in patients with bronchial asthma]. AB - Fifty-eight asthmatic patients (22 men and 36 women), aged 17 to 61 years, participated in this study. Thirty-two of them had atopic asthma. Exercise test and histamine inhalation challenge test were performed in all subjects. Patients exercised on a bicycle ergometer for 6-8 min. to increase the heart rate to 80% of predicted maximal heart rate. Twenty-three patients had a bronchospasm following exercise. The PC20 values in patients with exercise induced asthma (Me = 0.35 mg/ml) were significantly lower (p = 0.006) than in subjects without exercise induced asthma (Me = 1.89 mg/ml). There was low but statistically significant correlation between PC20 and delta FEV1 after exercise (R(s) = 0.32; p = 0.01). These results suggest that there are not only similarities but also differences in pathomechanisms of bronchial hyperreactivity to histamine and exercise-induced bronchoconstriction. The lack of statistically significant correlation between diurnal PEF variation and PC20 values suggest that the former parameter cannot be considered as an useful marker of nonspecific bronchial hyperreactivity. PMID- 10481522 TI - [Humoral immune response against A60 antigen from tuberculosis expectoration and the clinical and radiologic state of patients with with pulmonary tuberculosis]. AB - Serological tests available for diagnosis of tuberculosis can provide the valuable informations about host immune response to the mycobacterial infection. The aim of our study was to assess the correlation between clinical, radiological and bacteriological state of pulmonary tuberculosis patients and the serum concentration of IgG antibodies against A60 antigen of Mycobacterium bovis BCG. 110 active pulmonary tuberculosis patients--56 culture positive and 54 culture negative--were involved into the study. In all patients serum level of IgG ati A60 antibodies was measured with the use of A60-ELISA test. We found that bacteriologically confirmed pulmonary tuberculosis is associated with higher serum level of anti-A60 antibodies. Our study also showed the significantly higher levels of anti-A60 IgG in patients with cavitary and more progressive forms of this disease. We conclude that the greater antigenic challenge in progressive forms of pulmonary tuberculosis stimulates more expressed antibody response. PMID- 10481523 TI - [Clinical presentation and medical management in patients with idiopathic pulmonary fibrosis before diagnosis]. AB - The aim of the study was to find factors that may influence diagnostic process of idiopathic pulmonary fibrosis (IPF). Clinical presentation and medical management in the period before the diagnosis of IPF were assessed in 57 patients. There were 25 females and 32 males in the study group; mean age was 61.4 +/- 10.9 years. Mean duration of symptoms in the whole group was 18.4 +/- 16.0 months. The most common symptoms were dyspnea on exertion (96.5% of cases) and cough (84.2% of cases). 56 out of 57 patients (98.2% of cases) had crackles on auscultation of the lungs. In one third of the patients clubbing of the fingers was found. Mean FVC% of predicted was 77.2 +/- 19.8%, and mean DLCO% of predicted was 61.6 +/- 16.8%. The most common associated disease was ischemic heart disease, found in 43.9% of patients. Duration of symptoms in patients with ischemic heart disease was 25.7 +/- 18.8 months and was as twice long as in the rest of the patients, 12.8 +/- 10.5 months (p < 0.005). Prior the diagnosis of IPF 22 patients (38.6% of cases) did not any receive treatment for symptoms of interstitial lung disease. 31 patients (54.4% of cases) received antibacterial treatment (29 antibiotics and 2 anti-Tb drugs), although there were no clear signs of infection. CONCLUSIONS: 1. Associated ischemic heart disease may cause delay in establishing the diagnosis of IPF; 2. In the period before diagnosis of IPF over half of the patients were given antibacterial treatment without appropriate indications. PMID- 10481524 TI - [Noninvasive positive pressure ventilation in COPD patients with hypercapnic respiratory failure]. AB - The effects of noninvasive positive pressure ventilation (NIPPV) in COPD patients (pts) with hypercapnic respiratory failure were evaluated. The study group consisted of 19 COPD pts (16M, 3F, mean age 60 +/- 8 years) on LTOT for at least 6 month before study. Patients were enrolled in random order to group I, which continued LTOT and to group II, which started nocturnal NIPPV and continued LTOT. There were 12 pts in group 1 and 7 pts in group II. Two pts from the group did not tolerate NIPPV and were transferred to group I. To ventilate the pts we used portable, volume ventilators. Mean time of follow-up in group I was 23 +/- 13 months and 16 +/- 10 months in group II. During that time died 5 pts from 1 and 4 pts from group II. Differences between functional variables (FEV1, FVC, FEV1/VC, PaO2, PaCO2, pH, PEmax, 6MWD), dyspnea, number of hospitalizations and mortality in both groups were not statistically significant. In both groups progression of the disease (decrease of FEV1, worsening of hypoxaemia and increase of hypercapnia) was observed. NIPPV did not slow down progression of the disease. PMID- 10481525 TI - [The use of monoclonal antibodies in the detection of small cell lung cancer metastases in bone marrow]. AB - Expression of a number of antigens associated with small cell lung cancer (SCLC) have been proposed as a marker of malignancy and the diagnostic tool for the staging procedures and important prognostic factor. Since the bone marrow (BM) was described as a frequent site for SCLC metastases, we have decided to assess clinical importance of cancer cells detection in BM, using immunofluorescence with MAC-1, MAC-31, NSE and anti-Fucosyl-GM1 (PF3) antibodies. The group of 32 patients with SCLC was assessed using our panel of antibodies. Control group consisted of 5 patients with other malignancies (3 patients with malignant lymphoma, 1 with chronic lymphocytic leukaemia and 1 with non-SCLC). The study revealed no correlation between the expression of SCLC markers in patients BM and the cancer treatment outcome measured as a response for treatment, time to progression, and survival time, and no significant difference was found between the patients and control group. PMID- 10481526 TI - [A case of empyema after pneumonectomy caused by methicillin-resistant staphylococcus aureus infection treated successfully with local administration with vancomycin]. AB - A 55-year-old man underwent left pneumonectomy due to squamous cell carcinoma. Three weeks later bronchopleural fistula and pleural empyema with MRSA infection were recognized. Treatment was based on closed pleural drainage and antibiotic therapy. Initially patient was treated with trimethoprin-sulfamethoxazole and then vancomycin intravenously but empyema was not cured completely. Therefore repeated instillation of vancomycin into the empyema cavity was applied. After 6 days of treatment, culture studies of the pleural fluid became negative and drainage tube was removed 5 days later. We suggest that local administration of vancomycin is an effective method in postpneumonectomy empyema with MRSA infection. PMID- 10481527 TI - [Cutaneous tuberculosis and lymphatic nodes]. AB - 73-year-old woman with lesions in the right lung, fluid in the right pleural cavity, magnification of the left supraclavicular lymphatic nodes and ulcerations on the skin of the neck and chest is presented. Culture of 1 pus specimen received from the one of the skin ulcerations reveals the presence of Mycobacterium tuberculosis. Antituberculotic drugs (INH, RMP, EMB) were administered. Complete remission of the skin ulcerations and pleural exudate after 8 weeks and partial remission of the lesions in the lung and supraclavicular lymphatic nodes after 12 weeks of treatment was observed. PMID- 10481528 TI - [Treatment of cachexia in the course of lung cancer]. PMID- 10481529 TI - 40 years of the Polish Society of Pathologists--oral presentation at the 14th Congress of the Polish Society of Pathologists, Bydgoszcz, 10-12 September 1998. PMID- 10481530 TI - Clinico-pathological characteristics of colorectal cancer and serum anti-p53 antibodies. AB - The purpose of this study was to correlate the presence of p53 antibodies in sera of patients with colorectal adenocarcinoma with size, site and stage of the tumour, age and sex of a patient and the level of carcinoembryonic antigen (CEA) in the serum. p53 antibodies were detected using enzyme-linked immunoabsorbent assay (ELISA). Serum p53 antibodies were detected in 30 of 145 patients (21%), mostly in Astler-Coller stage B1 (28% of patients). No association was found between p53 antibody status in stage A+B1+B2 vs stages C1+C2+D (22% vs 19%) i.e. between patients without and with metastases to regional lymph nodes and/or distant metastases. Serum p53 antibodies were detected in 9 of 34 patients (26%) with tumour localised in the right part vs 21 of 109 patients (19%) with tumours in the left part of the colon and in 18 of 96 (19%) of patients with tumours localised in rectosigmoideum vs 12 of 47 (26%) with tumours in the remaining colon. There was no significant correlation between serum anti p53 antibody and CEA statuses. Increased level of serum CEA was seen in 46/145 (32%) patients. Patients with C1+C2+D stage cancers had high serum CEA level more frequently than did patients with A+B1+B2 stage tumours (44% vs 19% respectively, p < 0.001). Of 102 cases with normal CEA level, 19 (19%) were positive for anti p53 antibodies. These results together with the literature data [11, 20] indicate that approximately 27% CEA negative patients may have serum p53 antibodies. Therefore simultaneous assessment of serum p53 antibodies and CEA seems to be useful for monitoring high risk patients and for postoperative patient monitoring. PMID- 10481531 TI - Distribution, activity and concentration of cathepsin B and cystatin C in the wall of aortic aneurysm. AB - Intensity of immunohistochemical reaction to cathepsin B is stronger and to cystatin C weaker in the wall of aortic aneurysm than in normal aorta. Localization of cathepsin B and cystatin C in the aneurysm wall is different from that in the control aorta, but activity and concentration of cystatin C in the aneurysmal wall is lower than in normal aorta. The parietal thrombus of aneurysm also shows high activity of cathepsin B. The obtained results point to participation of cathepsin B in degradation of aneurysmal structural proteins and in enlargement of the aneurysmal size. PMID- 10481532 TI - Light and electron microscopic picture of atherosclerotic plaque in stable and unstable angina. AB - By presenting this series of 127 cases of coronary atherectomy the authors join the workers who study morphological differences between the atherosclerotic plaques in stable and unstable angina. Routine staining of formalin-fixed, paraffin-embedded material was completed by the detection of T lymphocytes, macrophages, mast cells, smooth myocytes and grown-in capillaries using monoclonal antibodies (DAKO), as well as by the immunofluorescent demonstration of fibrinogen in the plaques. The plaques derived from patients with unstable angina showed a higher incidence of mast cells (significant) and macrophages (insignificant). These cells render the plaque more susceptible to rupture or fissuring. There was also significantly more frequent and quantitatively more abundant permeation of the plaque by fibrinogen that raises the chance of thrombosis. These findings support the view that unstable angina correlates with the phenomena that favour the rupture of the plaque and thrombosis. Electron microscopy has not been used so far to study coronary atherosclerotic plaques. This material includes 15 plaques from stable and 18 plaques from unstable angina. A cover of fibrin and blood platelets is a regular formation on the surface and in the superficial layer of the plaque from unstable angina. It contributes to the "thrombotic proneness" of the coronary artery. These plaques also show abundant elastic fibres. This pattern corresponds to myo-elastic intimal hyperplasia ("intimal thickening") where the production of intimal elastin constitutes an essential phenomenon. Intimal thickening is interpreted as a preatherosclerotic event. The presence of elastin reflects an early stage of the development of the plaque. The plaque from stable angina shows abundant collagen fibres, which aggravate the lesion. PMID- 10481534 TI - Sclerosing hemangioma of the lung--benign sclerosing pneumocytoma. AB - Three cases of sclerosing hemangioma of the lung were studied. All developed in women aged 28, 32 and 59 years. They had been discovered on routine chest roentgenograms and then surgically removed. In two cases the tumours were solitary, in one case there were six tumours varying in size. The histology of this rare tumour and its various types as well as the results of immunohistochemical studies have been described, partly indicating its epithelial origin. PMID- 10481533 TI - The presence of Chlamydia pneumoniae in atherosclerotic plaques--a report of three cases of ischaemic heart disease. AB - The report presents three cases of ischaemic heart disease in which Chlamydia pneumoniae infections were detected first serologically, later the bacteria were shown in atherosclerotic plaques with electron microscopy, and finally C. pneumoniae strains were isolated from the tissues. PMID- 10481535 TI - The ultrastructural study of primary intracranial germ cell tumours. AB - We describe here ultrastructural and clinicopathological features of five primary intracranial germinomas. By electron microscopy, two major tumour components were defined as large, well differentiated tumour cells and non-neoplastic cells such as macrophages, astrocytes and lymphocytes. Nuclei of the tumour cells often presented irregularly contoured nuclear membranes with oval indentations and, occasionally, cytoplasmic invagination. Some of them constituted unusual conformational changes of nuclear membranes rarely described as intranuclear pockets. Desmosome-like intercellular junctions were observed in several neoplastic cells. Nucleoli were composed of a loose, fragmented nucleolonema whereas elongated, anastomosing and rope-like nucleolonemas, described previously as characteristic for germinomas were not seen. Typically, the cytoplasm contained glycogen particles. Most tumour cells had villous cytoplasmic projections sometimes intermingled with similar projections of macrophages. Scattered astrocytes typically containing abundant glial filaments were adjacent to primary tumour cells. PMID- 10481536 TI - Intelligent decision support in pathomorphology. AB - This paper presents a novel approach to computer-supported diagnosing based on microscopic images of histological sections. A method of extraction of textural feature is presented, which is in a sense complementary to the texture-based segmentation. The textural feature is obtained by tracing the process of image segmentation. For classification, a n2-classifier oriented to multi-class problems has been used. The paper presents also an empirical verification of the proposed approach on 700 microscopic images representing 14 classes of CNS neuroepithelial tumours, in which case an encouraging accuracy of classification on the testing set (70.6%) has been obtained. PMID- 10481537 TI - Primary localized cutaneous amyloidosis--lichen amyloidosus. A case report. AB - We report a case of primary localized cutaneous amyloidosis-lichen amyloidosus in a 55-year-old man. Immunohistochemistry using antibodies against cytokeratin and AL immunoglobulins revealed the presence of both components in amyloid foci located subepidermally, mainly in dermal papillae. The results of histochemical reactions confirm the keratin-derived nature of amyloid in primary cutaneous amyloidosis. PMID- 10481538 TI - [Herpes simplex encephalitis]. AB - Herpes simplex virus infection of the central nervous system is still a significant cause of morbidity and often mortality. Changes of central nervous system are results of primary infection or activation of latent HSV-1, HSV-2. Neurological deficits often follow encephalitis herpetica. The application of PCR is prompt and specific diagnosis of herpes simplex virus infections of the brain. Advances in treatment herpes simplex encephalitis with acyclovir have improved outcome. PMID- 10481539 TI - [Localization of ischemic lesions in the brain in relation to the main risk factors of stroke]. AB - The aim of the study was the correlation between the risk factors of stroke (arterial hypertension, diabetes mellitus, arrhythmias) and localization of ischemic lesion assessed by CT. The group consisted of 160 patients with ischemic stroke (70 male, 90 female), average age was 62.7 years. The group was divided into the subgroups depending on the localization and extension of ischemic lesion found on CT exam. The first group with cortical-subcortical lesions consisted of 100 patients. The second group with small lesions in deep brain structures consisted of 51 patients. The third consisted of 9 patients with both lesions: cortical subcortical and deep brain structures. 97 patients had history of HTN, 37DM, 47 arrhythmias. Occurrence of HTN was found to be significantly more frequent in the second subgroup. No significant correlation was found between localization of lesion and the diabetes and arrhythmias in remaining subgroups. PMID- 10481540 TI - [The comparison of the effectiveness of treatment of multiple sclerosis relapse with high doses of methylprednisolone, alpha-24 corticotropin and cyclophosphamide]. AB - The effectiveness of treatment was compared in groups of patients with definite multiple sclerosis (MS) given respectively: methylprednisolone, alpha-24 corticotropine and cyclophosphamide combined with corticotropine. Clinical improvement was assessed by means of the expanded disability status scale (EDSS). The significant results were noted in patients treated with methylprednisolon and corticotropine combined with cyclophosphamide (1.5 in EDSS) for the first time, when compared to subsequent treatment. Within the patients in a more advanced stage of the disease, undergoing subsequent treatment, those given corticotropine displayed the best improvement (1.5 in EDSS), but without statistical significance. The authors suggest methylprednisolone or corticotropine combined with cyclophosphamide are the most effective immunosuppressive treatment in the initial phase of MS. PMID- 10481541 TI - [Surgical treatment of thyroid disease in adolescents]. AB - Surgical treatment of goitre in children and teenagers under 18 is relatively rare and undertaken for strict therapeutic indications. It still arouses much controversy among surgeons, being loaded with a high percentage of complications, usually distant ones. The aim of the study was to analyse the surgical treatment of various goitre types under the age of 18 basing on 65 operations performed in the years 1994-1998. Operations for non-toxic and hyperactive nodular goitre, including Graves-Basedow disease, were predominant. Five cases of thyroid carcinoma were found. No serious complications were observed during and after the surgery. Our own material and the 5-year observation reveal that surgical treatment of thyroid diseases in children and adolescents carried out for definite indications by an experienced operating team is successful and causes only few complications. PMID- 10481542 TI - [Results of surgical treatment of adrenal gland tumors: own 15-year long experience]. AB - In the present communication we present the results of surgical treatment of tumours of the adrenal glands, basing on the material from our clinical department. During 1983-1997 at the Department of Clinical Urology Central Clinical Hospital of the Military University School of Medicine (CSK WAM) 27 patients (15 men, 12 women) with tumors of the adrenal glands were treated. Among the operated patients 3 were diagnosed with Cushing's syndrome, 3 with pheochromocytoma, 1 with Cohn's syndrome, while in the remaining 20 the tumours were hormonally inactive. Surgical treatment included 15 right, 10 left and 2 bilateral adrenalectomies. There was no intraoperational mortality. In all patients with hypersecretion, the symptoms disappeared after adrenalectomy. The size of tumours ranged 4-20 cm. In 3 patients there appeared a need for broadening of the size of surgical intervention--2 splenectomies and 2 nephrectomies and 1 resection of pancreatic tail--all due to regional infiltrations with tumorous tissues. During surgery there occurred 2 cases of iatrogenic pleuric injury. Postoperative complications included 1 case of pneumonia and 1 case of wound infection. In case of unilateral tumours of the adrenal glands the best method for surgery appears to be the lateral (lumbal) resection and in case of large tumours and/or bilateral tumours the most convenient appears to be the anterior (transperitoneal) resection. The most applicable method for diagnosis of tumours of the adrenal glands appears to be CT and NMR. PMID- 10481543 TI - [Changes of selected immunological parameters after cardiopulmonary bypass in postop period]. AB - The aim of the study was to evaluate the influence of changes of chosen immunological parameters on postoperative course patients after cardiopulmonary bypass operation. Complement components C3, C4 and immunoglobulins IgA, IgG, IgM were taken into account. The group consisted of 70 patients, 51 men at mean age 52.6 +/- 10.8 years and 19 women at mean age 50.7 +/- 11.0 years. All patients were operated in moderate hypothermia 26-32 degrees C with use of crystalloid cardioplegia. We used membrane oxygenators: Safe II (Polystan), Monolyth (Sorin), Maxima (Medtronic) and Bentley (Baxter). In 36 patients with multivessel coronary artery disease the internal thoracic artery and saphena vein grafts were performed. 27 patients underwent the valve prosthesis implantation procedure and 7 correction of the congenital heart dis-ease. The mean extracorporeal perfusion time was 127.5 +/- 51.0 min. The mean aortic cross-clamping time was 65.6 +/- 26.9 min. 6 blood samples were taken in the time periods called from 0 to 5: 0- before the operation, 1--right after the operation, 2--1 day after the operation, 3--3 days after the operation, 4--7 days after the operation, 5--14 days after the operation. All the immunological parameters were measured at the Technicon RA 1000 System device using plasma antibody serum of Behring Company. We compared two groups: 1) 21 patients extubated at operation day with 42 patients extubated at 1-th postoperative day, 2) 38 patients with postoperative organ failure with 32 patients without organ complications. The intubation time was shorter in patients with higher levels of C3 (to 7-th day) and C4 (at 1-th postoperative day). The postoperative organ failure were more frequently in patients with lower postoperative C3 (to 3-th day) and with lower C4 at 1-th postoperative day. The postoperative changes of immunoglobulins IgA, IgG, IgM were similar in patients with complicated and uncomplicated postoperative course. PMID- 10481544 TI - [Personality changes in children treated by the Ilizarov device]. AB - The treatment of children and youth by Ilizarov's method is very long. During this treatment there appear psychological problems which, on the one hand, are the effects of the features of personality but, on the other hand, they are the effects of emotional lability which is characteristic of adolescence. This paper basing on analysis of questionnaires and chosen personality features attempts to find correlation between these features and the effects of treatment. The obtained findings suggest that patients who are treated by Ilizarov's method should undergo psychological therapy. PMID- 10481545 TI - [Application of pentoxiphylline to treatment of leg ulcers: a Polish multicenter study]. AB - Aim of the study was to evaluate the efficacy, tolerance and safety of treatment of leg ulcer with pentoxiphylline administered orally in daily dose of 1200 mg divided for 3 doses for six months. The study was carried out in 19 centres in Poland. The results of clinical and laboratory (HGB, WBC, PTL) examinations made before, during and after treatment were documented in individual case-records. Pentoxiphylline was administered to 184 patients (124 F and 60 M) aged 22-86 (mean-61.6) yrs. One hundred and thirty-five patients had non-arterial ulcers, 37 partly arterial and 12-arterial. The duration of the disease was 1 to 1620 (mean 158) weeks. Resting pain was reported by 131 patients. In 9 persons the treatment was stopped after 1-2 months; in 6 (3%) of them due to side effects. In 175 patients pentoxiphylline was administered as follows: 1200 mg daily for 6 months in 92 cases, 1200 mg daily for 2-5 months--in 58, 1200 mg daily for 3-5 months and 800 mg daily for next 1-3 months--in 12 cases, and 800 mg daily for 1-6 months--in 13 cases. In 161 (92%) cases a beneficial effect was achieved including complete healing of the ulcers in 79 (45%) or improvement in 82 (47%); in 14 (8%) there was no improvement. The resting pain was significantly reduced in 114 (87%) patients within 1-24 weeks and completely disappeared in 69 (53%) patients within 2-20 weeks of treatment. No significant abnormalities in the results of laboratory tests were found. Transient side effects of the therapy appeared in 11 patients but they did not require administration of the drug to be discontinued. The study showed that pentoxiphylline is a effective, well tolerated and safe drug in the treatment of leg ulcers. PMID- 10481546 TI - [The efficacy of subcutaneous sumatriptan for the treatment of migraine attack]. AB - Sumatriptan is a novel drug for the treatment of acute migraine attacks. The aim of this study was to assess the efficacy of a 6 mg s.c. Sumatriptan in the early acute treatment of migraine. Patients were recruited with a current history of migraine with and without aura. 58 patients treated 1 migraine attacks at home using an autoinjector with a dose 6 mg of sumatriptan. Patient assessed headache severity and other migraine symptoms 1 and 4 h after treatment. 55% patients reported headache relief after 1 h and 77% patients after 4 h. Adverse events were reported by 11% of patients. In conclusion, sumatriptan were effective and well tolerated in the acute treatment of migraine. PMID- 10481547 TI - [MRI of traumatic superficial temporal artery aneurysm. Case report]. AB - A case of 14-years-old girl with superficial temporal artery aneurysm is presented. She sustained head injury six weeks before the first symptom. Presence of the recent blunt trauma in confrontation with MRI lead to establishing of the final diagnosis. It must be considered in the differential diagnosis of masses located in the temporal fossa. Surgical resection of the involved segment of the vessel is indicated in the cases without the need of reconstruction. The result of treatment in our case was excellent and all symptoms have resolved. The aneurysmal character of the lesion was confirmed by histopathological examination. PMID- 10481548 TI - [Hyperkalemic periodic muscle paralysis in primary adrenocortical insufficiency. A case report]. AB - 20-years old man was admitted to the hospital because of a few paroxysmal muscle paralysis episodes with pronounced periodic hiperkalemia (maximal 9.8 mmol/l). The first episode was preceded by a very hard physical effort. Primary adrenal insufficiency was recognised on the basis of clinical, humoral and biochemical examinations. There was elevated ACTH and a very low serum level of cortisol and aldosterone. There was slight metabolic acidosis and hiponatremia. The patient was treated with hydrocortisone and fludrocortisone acetate (Cortineff) with positive result. During one year of this therapy his general condition was very good and clinical, humoral and biochemical signs (except of metabolic acidosis) resolved. Neurological symptoms and a very high serum kalium level were the most important signs of primary adrenal insufficiency in the presented case. PMID- 10481549 TI - [Evaluation of quality of life in patients after stroke]. AB - The investigations concerning quality of life after stroke provide an alternative approach in comparison with traditional evaluation of the patient's condition focused only on neurological deficits. The full picture of the patient after stroke should take into account not only locomotor inefficiency, but also cognitive deficits, emotional disturbances and limitations in performing one's social functions. The study present attempts to define health related quality of life and reviews questionnaires devised for the examination of post-stroke patients. Despite a wide variety of methods used, the evaluation of all the areas of the patient's life after stroke seems impossible using only one research tool. There is also an urgent need to develop a uniform system of evaluation, appropriate for Polish cultural standards. PMID- 10481550 TI - [Possibilities of objective evaluation of treatment and rehabilitation effects after the stroke]. AB - The development of scoring scales has been initiated because of the need of documentation of results as well treatment as rehabilitation of stroke hemiplegic patients. Directly after onset at stroke unit there is demand for defining of prognosis. In rehabilitation the interest in stroke scales has been aroused from the need of evaluation of functional state (activities of daily living), disability, handicap, quality of life, statement of capacity for work or need of home-care. For evaluation of outcome after rehabilitation the most usefull seem to be functional scales (ADL) and quality of life measures (QOL). It is also suitable to mention the scales that evaluate motor function--those functions are the most important task in poststroke rehabilitation. PMID- 10481551 TI - [The role of agonists of serotonin receptor 5HT1B/D in pathogenesis and treatment of migraine attacks]. AB - The role of 5HT receptor agonists in pathogenesis of migraine are presented in the review in therapeutic aspects. Serotonin has been documented in several studies as an important mediator in migraine reactions and its receptors has been classified from 5HT1 to 5HT7. Serotonin receptor agonists are able to control and inhibit migraine attacks, however their mechanism of action is known only partially. PMID- 10481552 TI - [Lyme disease vaccine: hopes and concerns]. AB - The vaccine against Lyme disease for last decade became an important issue for many researchers. Recently two groups of scientists published data of their trials carried out on humans. The vaccine which was given to volunteers consisted of recombinant outer surface protein (Osp)A. These people were observed for two years when approximately 79% of them developed resistance to Borrelia burgdorferi infection. These data were evaluated by FDA experts who recommended vaccine for approval. Main concerns of experts are: short duration of observation of vaccinated people, no possibility to vaccinate children, problematic vaccination people with undiagnosed Lyme disease. However the moment when effective and safe vaccine against Lyme disease will be available seems to approach. PMID- 10481553 TI - [Practical aspects of iodine-131 therapy of hyperthyroidism]. AB - The paper overviews indications, contraindications, principles of dose determinations and management during and post radioiodine therapy of hyperthyroidism. It is specially focused on the differences in preparing and treating of patients with Graves and Plummer or Goetsch diseases. PMID- 10481554 TI - [Therapeutic advances in epilepsy]. PMID- 10481555 TI - [Gastroesophageal reflux]. PMID- 10481556 TI - [Functional deterioration secondary to hospitalization for an acute disease in the elderly. An analysis of its incidence and the associated risk factors]. AB - OBJECTIVE: To determine the incidence of functional impairment at hospital admission and after hospital discharge, and to identify those factors that could be predictors of such impairment in a cohort of elderly people with high level of independence in the basic activities of daily living (BADL). DESIGN: Epidemiologic, longitudinal and prospective study. SETTING: Acute unit in a geriatric department. SUBJECTS: The follow-up population was made up by 150 subjects older than 65 years, admitted on account of acute medical conditions from December, 15th, 1995, to December, 15th, 1996, who were independent for walking and independent or with slight dependence for BADL. STUDY PARAMETERS: All patients were evaluated at 48 hours after admission and one month after discharge by means of a protocol including sociodemographic, clinical, attending, functional and mental parameters. RESULTS: The incidence of functional impairment at hospital admission was 70.66%; such impairment persisted one month after discharge in 29.3% of subjects. The most involved activities related to motility. In the analysis of crude data, the following parameters showed association with functional impairment risk one month after discharge: female sex (OR: 2.5), sensorial organ pathology (OR: 2.6), hospital stay longer than 15 days (OR: 7.2), and Lawton Index score (3.8 +/- 2.6 in impaired patients vs. 4.9 +/- 2 in non impaired patients; p < 0.01). Barthel Index score at admission lower than 60 (OR: 9.5), Cognitive Miniexam score lower than 28 (OR: 4.1), Informant Test score higher than 84 (OR: 2.5) and Geriatric Depression Scale score 9 (OR: 3.1). In the logistic regression model, the following parameters remained as predictors of impairment: sex (OR: 3.3), days of hospital stay (OR: 2.3) and Barthel Index score at admission (OR: 6.1) and Cognitive Miniexam score (OR: 2.7). CONCLUSIONS: For our population, sex, hospital stay days, functional impairment in Barthel Index at admission and Cognitive Miniexam score parameters seem to behave as independent variables that can predict functional impairment risk for BADL one month after discharge. These parameters could be useful to select groups of elderly people with impairment risk, who could benefit from specialized interventions leading to prevent/decrease functional impairment secondary to acute disease and hospitalization. PMID- 10481557 TI - [The epidemiological surveillance of acute poisoning in 1997 (a study of 1140 cases from the area south of the Madrid Community)]. AB - Following a line of work we studied 1,140 acute poisoning (AP) attended at the Internal Medicine Emergency Department at 12 de Octubre Hospital, Madrid, in 1997. The incidence increased up to 157/100,000 inhabitants. Mean age was 36 years (SD: 15 y), median, 32 years. Self-inflicted AP: 1,052 cases (92%); the suicide attempt was the most common type (509, 48%). Among males, the alcoholic intoxication (332, 59%) predominated. Among accidental AP (88 cases), 90% occurred at home. Poisons: drugs. Drugs were used for 78% of suicide attempts. The relative incidence of benzodiazepines increased (47%) and that of antidepressive drugs decreased (11%). A remarkable increase in the "other drugs" group was noted, as well as the association of drugs and non-drugs (10%). Alcohol use increased significantly (249 cases more than in 1994), as well as drugs although to a lesser extent, breaking the trend observed in the last few years. Cocaine is now the most common (among women heroin is still the leading cause); 5 AP were caused by synthesis drugs (extasis, MDMA) and an increase was observed with "other drugs". ANTECEDENTS: suicide attempts: depression (30%) and previous attempts (19%); ethylism: alcoholism (40%); AP with other drugs: drug abuse (52%). Admitted to ICU: 1.4%. The mortality rate decreased to 0.08%. PMID- 10481559 TI - [Solitary fibrous tumors of the pleura]. AB - Seven cases of solitary fibrous tumours of the pleura are here reported, formerly named benign fibrous mesotheliomas. Histologically, these tumours are formed by fusiform cells with a minimal cellular pleomorphism and absence of mitosis. These tumours cause scarce symptoms which are usually diagnosed as a casual finding when a chest X-ray is performed on account of other cause. Treatment should be surgical, with removal of all tumoral mass, thus avoiding its growth and the possibility of the reported degeneration towards malignancy. PMID- 10481558 TI - [The palliative treatment of malignant esophageal stenosis and fistulae with self expanding metal prostheses]. AB - OBJECTIVE: To evaluate the efficiency of self-expanding metal stents (SES) for the palliative treatment of malignant oesophageal strictures and fistulas. MATERIAL AND METHODS: Fifty non-operable patients with malignant oesophageal strictures and/or fistulas were treated with SES placement. All patients had dysphagia, the mean degree for the studied group being 3.36. Ten patients presented associated tumoral fistula. Uncoated SES were placed for simple stenosis cases and coated SES for cancers with associated fistulas. RESULTS: A improvement in dysphagia was obtained in 47 patients (94%). The mean degree of dysphagia after the procedure was 1.12. In nine patients (90%) the fistula closed with resolution of the associated clinical picture. Complication were observed in six patients (12%), the perioperative mortality rate being 2%. During follow-up, twelve patients (25%) had malfunction of the stent, which was successfully treated in eight patients. CONCLUSION: SES are efficient for the palliative treatment of malignant oesophageal strictures and/or fistulas, but not commonly re-interventions are necessary to maintain a long-term functionality. PMID- 10481560 TI - [Macroamylasemia in the differential diagnosis of acute pancreatitis]. AB - The presence of macroamylasemia should be considered in the differential diagnosis of hyperamylasemia associated with acute abdominal pain, with suspect of pancreatitis. This uncommon and poorly known abnormality is defined as a blinding of serum amylase to certain proteins forming a circulating macrocomplex which prevents renal clearance of serum amylase. Two clinical cases as well as a review of this rare entity are here reported. PMID- 10481561 TI - [The risks of exposure to ionizing radiation during pregnancy and of the exposure of the gonads before conception]. PMID- 10481562 TI - [Volume-reduction surgery for pulmonary edema]. PMID- 10481563 TI - [Vascular risk factors and cognitive change in the elderly]. PMID- 10481564 TI - [Pulmonary nodules in a patient with von Recklinghausen's disease]. PMID- 10481565 TI - [Unilateral hypoventilation in the adult patient]. PMID- 10481566 TI - [Acute abdomen in a young woman with acute nonlymphoblastic leukemia]. PMID- 10481567 TI - [The cauda equina syndrome and a sacral lithic mass]. PMID- 10481568 TI - [The treatment of asthmatic crisis]. PMID- 10481569 TI - [Septic oligoarthritis due to Streptococcus pneumoniae]. PMID- 10481570 TI - [The black thyroid syndrome. A case report]. PMID- 10481571 TI - [The efficacy of sequential treatment with amphotericin B-fluorocytosine and fluconazole in cryptococcal meningitis not associated with AIDS]. PMID- 10481572 TI - [Amyloid goiter as the beginning symptom of amyloidosis in a patient with sex linked agammaglobulinemia]. PMID- 10481573 TI - [Torpid pneumonia secondary to the aspiration of a phonation prosthesis]. PMID- 10481574 TI - [Hyperthyroidism and atrial fibrillation]. PMID- 10481575 TI - [A new classification for the third millenium. When and until when?]. PMID- 10481576 TI - [Complications associated with central venous catheters in patient with hematologic neoplasms or hematopoietic transplants]. AB - OBJECTIVE: Nontunneled central venous catheters (CVCs) have been found an excellent cost-effective alternative to tunneled CVCs with comparable durability and safety when managed by a specialized team. The objective of this study was to evaluate the complications related with a nontunneled polyurethane CVC in a medium-size hospital without such dedicated services. PATIENTS AND METHODS: A representative sample of 82 cancer patients with 123 nontunneled CVCs inserted at our centre were followed up and evaluated clinically and microbiologically. Insertion and care were performed by the medical and nursing teams. RESULTS: The mean duration of the catheters in place was 28.2 days. Eleven mechanic complications (8.9%) were observed. We had a total of 3,380 days of catheter use with an infection rate of 0.86 per 100 catheter-days. Staphylococcus coagulase negative was the most common microorganism isolated. Local and systemic infection showed a different pattern of incidence, early after insertion and a month later respectively. Male sex and neutropenia at catheter removal were the only risk factors for bacteremia while receiving antibiotics at insertion date was a protecting factor. Age, number of lumens, insertion difficulty or patient diagnosis were not related with infection risk. CONCLUSIONS: Contamination at catheter insertion clinical or manipulation must be avoided especially when a neutropenia period is expected. A highly trained team working under rigorous guidelines is an important factor for optimal clinical and economic results with nontunneled CVCs. The cost of a specialized infusion team may well be below the price of poorly maintained catheters. PMID- 10481577 TI - [Infectious and non-infectious complications of tunneled central catheters in hematologic patients]. AB - PURPOSE: Long-term therapy of haematology patients has been facilitated by permanent indwelling central venous catheters. We performed a retrospective study to compare the problems occurring with a externalized catheter (Hickman) versus a totally implanted port catheter. PATIENTS AND METHODS: A total of 171 catheters were placed to 139 haematological patients, 77 patients with Hickman catheters and 94 with totally implanted port catheters. We review our experience in order to identify factors associated with complications. RESULTS: Pneumothorax occurred in one of 171 of the percutaneously placed devices. Other early complications were hematoma 13, and catheter migration out of the vascular tree 8. Late complications included malposition (5.8%), thrombosis (2.9%), septic thrombosis (1.7%) and most notably infection (38.5%). 62 of 77 patients with Hickman catheters developed catheter-related infection (hazard rate infection 7.1/1000 days) compared with 53 of 94 patients with implanted port catheters (hazard rate infection 1.5/1000 days, p < 0.001). Most of infections that occurred were caused by gram-positive organisms but the gram-negative organisms infections resulted in a significantly higher rate of treatment failure and recurrence. A total of 72 catheters were removed of the central line: 36 for infection. CONCLUSION: We found a significantly increased incidence of catheter-related infection in patients with Hickman catheters. We also observed that the use of intravenous antibiotic prophylaxis prior to catheter insertion did not appear to be beneficial and thrombocytopenia at this moment was a factor in the development of hematoma. The infections due to coagulase-positive staphylococci can be treated successfully without removal of the catheters. However in catheter-related bacteremia due gram-negative organisms there is a chance that the bacteremia will recur if the catheter is not removed. PMID- 10481578 TI - [In vitro and in vivo studies of murine erythropoietic recovery after treatment with cyclophosphamide]. AB - OBJECTIVES: The aim of this study is to analyse functional changes in murine erythropoietic tissues over 20 days post cyclophosphamide (CY) treatment. The project was focused on the capability of femoral and splenic erythropoietic responsive cells (ERC) to proliferate with human recombinant erythropoietin (rh Epo) stimulation after cytotoxic treatment. MATERIALS AND METHODS: CF-1 mice were injected i.p. with CY (200 mg/Kg). Individual lots were studied at 0, 2, 5, 7, 10, 14, 17 and 20 days post cytotoxic treatment. Haematologic parameters [packed red cells (PRC) reticulocytes, white blood cells] were scored. Erythropoietic differentiation was assessed by the 59Fe uptake assay and the proliferative profiles of erythropoietic progenitors were determined by 3H-thymidine incorporation assay with several doses of rh Epo (0-250 mU/mL). Total and differential cellularities were scored in bone marrow (BM) and spleen. RESULTS: A drastical decrease of total nucleated BM cells was noticed at 2 days post CY, although cellularity was restored by the 7th day. Spleen, however, failed in showing significant decrease. The maintenance of PRC was achieved through a deep erythropoietic reorganization. 59Fe uptake revealed changes in erythroid differentiation in both tissues. Spleen maturative contribution to whole erythropoiesis was always less than medullary supply, except on day 10 post CY when a transient compensatory red cell contribution was noticed. Proliferative assays revealed that erythropoietic recovery in BM post CY was delayed in comparison with myelopoietic restoration. Splenic erythroid proliferative pattern correlated with differentiation data. CONCLUSIONS: Myelopoietic and erythropoietic progenitors showed different recovery patterns post CY administration in BM and spleen. Medullary hemopoietic lineages restoration described a particular sequence: myelopoiesis restitution was previous to the erythroid one. Medullary erythropoiesis occurred without drastic changes in erythropoietin sensitivity while the spleen showed a transient dramatic increment on 10 days post CY red proliferation. Experimental data strongly suggest that erythropoietic recovery after CY insult mainly depends on microenvironmental regulations rather than on hormone titers. PMID- 10481579 TI - [Immunophenotyping of acute lymphoblastic leukemia in Mexican children]. AB - PURPOSE: To analyse the immunophenotype of leukaemic cells in a group of children diagnosed of lymphoblastic leukaemia in order to assess the frequency of the different immunologic subtypes. PATIENTS AND METHODS: In the period comprised between APR 1987 and MAR 1995, 402 Mexican children were studied in a prospective way. Conventional immunological markers were used, either associated to or specific for B, T, myelo-monocytic or megakaryocytic-platelet cell populations. RESULTS: Five major immunologic subtypes were disclosed, showing a series of specific surface markers: null-ALL, 5%; early pre-B, 7.5%; common, 74.6%; B-cell, 3.5%, and T-cell, 9.4%. A net predominance of B-cell precursor CD10- ALL was found in children under one year of age, and of CD10+ B-cells beyond that age. Although there was only slight predominance of male sex, the prevalence of B and TALL in males was not confirmed. CONCLUSIONS: These results show that the incidence of the different immunologic subtypes of lymphoblastic leukaemias and their distribution according to age and sex are closely similar to those reported among Caucasians in other parts of the world. PMID- 10481580 TI - [Erythrocyte glutathione reductase in pregnant women with hemoglobin AA and AS]. AB - PURPOSE: To evaluate the catalytic activity of the red cell enzyme, glutathione reductase (GR) in pregnant women with Hb AS and with Hb AA, and in a group of non pregnant women with Hb AA, as well as the relationship of GR deficiency with Hb S. PATIENTS AND METHODS: The catalytic activity was determined in presence and absence of FAD by means of a modified Long and Carson technique. 59 pregnant women with AS and 33 with AA phenotypes were studied. RESULTS: Differences were found in the enzyme's catalytic activity with and without FAD, both in pregnant women with Hb AS (mean values 37.17 nka/g y Hb in whites and 42.84 nkat/g HS in afro people) and in those with Hb AA, and also in non-pregnant women with Hb AA. A high frequency of GR deficiency was found in all groups due to an insufficient riboflavin supply in diet. CONCLUSION: A correlation between GR deficiency and Hb S could not be demonstrated. The coefficient of activity of red cell GR shows a tendency to increase in pregnancy due to certain riboflavin deficit of diet. PMID- 10481581 TI - [Detection of PNH clones using flow cytometry in aplastic anemia and paroxysmal nocturnal hemoglobinuria]. AB - PURPOSE: To detect and quantify by flow cytometry (FC) PNH clones in paroxysmal nocturnal haemoglobinuria (PNH) and aplastic anaemia (AA) patients. PATIENTS AND METHODS: We have performed a flow cytometric analysis to determine the granulocyte expression of CD55 and CD59 from 29 patients with AA and 11 patients with PNH. RESULTS: In the 11 PNH patients the study showed 58 +/- 34% and 56 +/- 32% (mean +/- SD) CD55(-) y CD59(-) granulocytes. A good correlation was found between the results of FC and haemolysis. The follow-up study showed PNH clone progression in one case and stability in 5 cases. Among 11 AA patients studied at diagnosis, two presented a population of CD55(-) granulocytes (14% and 48%) with CD59 normal, this defect disappeared in both patients after immunosuppressive therapy. The FC study revealed PNH clones in 7 cases among the 26 analyzed after treatment (23 with ATG and/or CyA), in 3 cases with negative Ham's test (in two this became positive 6 and 12 months later). The mean values obtained in these 7 patients with PNH-AA syndrome were 26 +/- 15% y 36 +/- 30% (mean +/- SD) CD55(-) and CD59(-) granulocytes. The median time from diagnosis to detection of PNH phenomenon was 83 months. In the follow-up study, 4 cases had stability, one case had a decrease and one a progression of the abnormal clone. In a retrospective analysis, among the 7 patients with PNH-AA syndrome, 5 had a partial response after the initial treatment. CONCLUSIONS: The FC on granulocytes is a useful method to diagnose and characterize PNH. This test is good for early detection of PNH clones in AA patients at initial diagnosis and in long term survivors. In both diseases it permits measuring the extent of the abnormal clone and its follow up. The extent of the defect is more related to haemolysis than the haematopoietic deficiency. PNH development seems to be more frequent in AA patients with incomplete response after immunosuppressive therapy and in some cases the defect could be latent at the time of diagnosis. PMID- 10481582 TI - [Binding of the antileukemia drug Escherichia coli L-asparaginase to the plasma membrane of normal human mononuclear cells]. AB - OBJECTIVE: To demonstrate that the enzyme L-asparaginase from Escherichia coli (EcA) binds to the plasma membranes of normal human lymphocytes and monocytes. MATERIAL AND METHODS: Lymphocytes and monocytes were isolated from heparinized blood samples which came from healthy volunteer donors. The cells were incubated with EcA to detect a possible binding of the enzyme to the mononuclear cells by indirect immunofluorescence using confocal microscopy. Meanwhile, ultracentrifugation was used to obtain the erythrocyte ghost microsomal fraction (P100) which was then analyzed by Western blotting to determine if EcA binds the lipid bilayer unspecifically. For the immunoassays, monospecific polyclonal antibodies were obtained from ascitic tumors developed in mice immunized with commercial L-asparaginase. RESULTS: EcA bins the lymphocyte and monocyte plasma membranes. In monocytes, there occurs a capping phenomenon, that is, the accumulation of fluorescent marker in one region. The image analyzer highlights it clearly at a depth of 3.8 microns. This binding would be unspecific, that is, there is no mediation of a specific receptor that binds EcA. This arises from the ability of the enzyme to bind to the membranes of erythrocyte ghost, as evidenced by the ability of the molecule to associate with a hydrophobic medium. The antibodies against EcA obtained from ascitic tumours developed in mice do not show cross reactivity with Na+/K+ ATPase, aspartate aminotransferase, nor with extracts of blood cells, which would make it a specific tool for the detection of EcA in whole cells and in homogenates electrotransfered to nitrocellulose membranes. CONCLUSION: L-asparaginase from E. coli behaves as a lipoprotein due to its ability to insert itself into hydrophobic environments, in which it resembles an isozyme present in T. pyriformis. The binding of this enzyme to lymphocytes and monocytes, demonstrated in this work, would permit the modification of the antileukemic treatment injecting doses of EcA bound to patient's own isolated immune cells. PMID- 10481583 TI - [Molecular genetics of the thalassemias in Argentina]. AB - PURPOSE: Was to establish the molecular genetics of thalassemias in Argentina. PATIENTS AND METHODS: Genomic DNA was amplified by PCR and six point mutations in the beta-globin gene were investigated by Dot Blot hybridization using oligonucleotide probes. The most frequent alpha-thalassemia deletions were studied by Southern Blotting. Patients were distributed in 4 groups: a) 109 beta thalassemic carriers; b) 15 thalassemia major patients; c) 2 thalassemia intermedia patients and d) 14 probable alpha-thalassemic carriers. RESULTS: The distribution of mutated alleles in the group a) was: IVS-1 nt 1: 13.76%, IVS-1 nt 6: 7.34%, IVS-1 nt 110: 23.85%, codon 39: 39.45%, IVS-2 nt 1: 3.68% e IVS-2 nt 745: 1.83%, 10.01% could not be determined with the probes used; in the group b) the allelic distribution was similar and the compound genetic genotype were predominant related to homocygous ones; in the group c): we confirmed the presence of one beta-thalassemia mutation and a alpha gene triplication (alpha alpha alpha) in the 2 patients studied. The alpha-thalassemia character was confirmed in 8 patients of the group d) (6 had -alpha 3,7/alpha alpha genotype and 2,-alpha 3,7/-alpha 3,7 genotype). CONCLUSIONS: This study indicates that the analysis of 6 mutations in the beta-globin gene and the alpha-globin gene deletions are an effective strategy to identify thalassemias in Argentina. PMID- 10481584 TI - [Hemorrhagic complications from anticoagulant treatment: analysis of predictive risk factors]. AB - Anticoagulant therapy has shown its efficacy in the prevention of thromboembolic complications but it is not devoid of bleeding complications. Although the thromboembolic risk of some cardiac diseases may be extrapolated from well organized clinical trials, the risk of bleeding complications should be determined in the context of the environment in which it is carried out. OBJECTIVE: To determine the complications of the patients in anticoagulant therapy, in our environment, and to analyse the of risk factors. PATIENTS AND METHODS: We have studied the complications suffered by 300 patients who underwent anticoagulation for cardiac diseases, between March-94 and March-96 retrospectively. We have classified the complications in two groups: a) Fatal or intracranial with sequelae. b) Those requiring hospitalization and/or transfusion. Univariate and multivariant analyses were conducted to identify predictors of complications, including the following factors: age, sex, diabetes, hypertension, length of therapy, distance from our Center to their place of residence, INR (> 3 vs 2 to 3) and number of drugs associated with Acenocoumarol (> or = 3 vs 2 or less). RESULTS: During the follow-up 24 patients died due to non haemorrhagic complications; 3 left the treatment on their cardiologist recommendation; 2 moved their place of residence and 1 was lost in the follow-up. Of 270 remaining 3 (0.55/100 patients-year) had complications of group a and 21 (3.88/100 patients-year) of grub b. INR > 3 and multiple medications were shown as predictors of complications when including some of the complications considered. CONCLUSIONS: 1) During two years of follow-up 1.1% (0.55/100 patients year) of patients in anticaogulant therapy had bleeding complications resulting in death or neurological sequelae. 2) When including some of the complications considered the percentage rises to 4.44/100 patients-years. 3) Although there were no differences statistically significant, INR > 3 and polymedication have been found as predictors risk factor. 4) Patients with INR of 2 to 3 and non polymedicated presented a low risk of bleeding complications (1.66/100 patients year). PMID- 10481585 TI - [Antiphospholipid-protein antibodies. Effects on the endothelium-platelet interaction]. PMID- 10481586 TI - [Accreditation and certification of clinical laboratories]. PMID- 10481588 TI - [Review of informed consent for transfusion]. PMID- 10481587 TI - [Fraud against the Spanish MRI? And now what, four years later?]. PMID- 10481589 TI - [Evaluation of the leuko-reductive efficacy of single-donor erythropheresis procedures]. PMID- 10481590 TI - [Anti-HB-core antibodies as a surrogate marker in blood donors]. PMID- 10481591 TI - [Erythrocyte allotransfusion and venous thromboembolic disease]. PMID- 10481592 TI - [Primary hepatic non-Hodgkin's lymphoma]. PMID- 10481593 TI - [Burkitt leukemia as the presenting form of acquired immunodeficiency syndrome. Description of 2 cases]. PMID- 10481594 TI - [Lymphoma of the mandible: presentation of 2 cases]. PMID- 10481595 TI - [Severe esophageal stenosis secondary to the administration of chemotherapy in a patient with acute myeloblastic leukemia]. PMID- 10481596 TI - [Antifibrinolytics as a treatment for severe factor XI deficiency in laryngeal surgery]. PMID- 10481597 TI - [Correction]. PMID- 10481598 TI - [The Procera Allceram all-ceramic system. The clinical and technical laboratory aspects in the use of a new all-ceramic system]. AB - The aim of this article was to present the CAD/CAM-assisted Procera system as a new all-ceramic full-coverage crown system. New technologies need to be evaluated technically and clinically to be able to fulfill the requests for aesthetics, physical strength and biocompatibility when restoring the anterior and posterior region. The Procera AllCeram system offers the opportunity to fabricate densely sintered, high-purity, tooth-colored copings. Mechanical properties of the Procera alumina coping, indications and contraindications and additional applications are discussed. The clinical and technical procedures and any specific limitations and peculiarities are summarized and documented with clinical cases in a step-by-step illustration. PMID- 10481600 TI - [In Process Citation] PMID- 10481599 TI - [Primary non-Hodgkin lymphoma of the mandible--a case report]. PMID- 10481601 TI - [The genome of Bacillus subtilis and the features of its genes]. PMID- 10481602 TI - [Formation, structure and germination of the spore of Bacillus subtilis]. PMID- 10481603 TI - [Signal transduction, competence development and protein secretion in Bacillus subtilis]. PMID- 10481604 TI - [Molecular mechanisms and biological roles of apoptosis and inflammation induced by Fas ligand]. PMID- 10481605 TI - [Structure and function of cellulolytic complex "cellulosome"]. PMID- 10481606 TI - [Translational control of maternal mRNA by poly(A) tail elongation and cap ribose methylation]. PMID- 10481607 TI - [Impact of the complete genome sequence on biology of Bacillus subtilis]. PMID- 10481608 TI - [SPM, a method for isolation of CpG islands: application of stability of partly melted molecules]. PMID- 10481609 TI - [Thermal lens microscopy and single molecule determination of non-fluorescent molecules]. PMID- 10481610 TI - Induction of arthritis with Mycoplasma hyosynoviae in pigs: clinical response and re-isolation of the organism from body fluids and organs. AB - The objective of this investigation was to study the pathogenesis of experimental Mycoplasma hyosynoviae arthritis in pigs. The experimental inoculations were designed to provide information about systemic spread, the persistence of subclinical infection, and the length of time for which the mycoplasma is cultivable from synovial fluids and other tissues. In this article we report on the clinical response to infection and the results obtained from re-isolation attempts. In three inoculation experiments with M. hyosynoviae, clinical arthritis was produced by intravenous and by intranasal exposure as well as by pen-contact in 12 out of 23 exposed pigs. The infection was transmitted from persistent carrier pigs to non-infected pigs by pen-contact. The incubation period until development of clinical arthritis was 4-9 days for all routes of exposure, and the symptoms were of variable severity. In half of the cases the onset was acute and the lameness severe, typically involving the hindlegs and with affected pigs assuming a dog-sitting position. A systemic phase was found in the majority (86%) of the pigs. However, the infection was in two cases established in the tonsils without detection of a systemic phase. An apparent persistent infection of the tonsils became established in all the pigs. M. hyosynoviae spreads via the blood to different organs from which it could be re isolated during the acute phase of the infection. In general, M. hyosynoviae was recovered from joints from day 3 until day 21 post-exposure, but longer persistence of viable mycoplasmas in joints or regional lymph nodes in the chronic phases of the infection appeared to have taken place in a few pigs. PMID- 10481611 TI - Intrauterine growth retardation as a cause of perinatal mortality in Japanese black beef calves. AB - Perinatal death is one of the major causes of calf mortality in Japanese Black beef herds. A series of experiments were carried out to determine causes of perinatal calf mortality in a region of northern Japan. An aetiological survey revealed that the incidence of perinatal mortality in 6475 calves was 4.5% and the majority of neonatal deaths were caused by weak calf syndrome. The weak calves delivered after a normal gestation period had a significantly lower body weight at birth than normal calves (P < 0.01), indicating growth retardation in the uterus during pregnancy. Haematological and histopathological examinations showed that the weak calves had anaemia characterized by a significant decrease of blood cell values and bone marrow hypofunction. The anaemia due to bone marrow hypofunction presumably caused intrauterine growth retardation of the foetus. Dams delivering the weak calves showed significantly lower serum concentrations of estrone sulphate during late pregnancy than those with normal calves (P < 0.01), indicating foeto-placental dysfunction as a possible cause of growth retardation. Effects of sires as well as the maternal family on the incidence of neonatal death and on the rate of weak calves in dead neonates were found in the survey. Intrauterine growth retardation associated with anaemia may be the most important cause of weak calf syndrome in Japanese Black beef calves. The bull and maternal family may also have an influence on the incidence of neonatal death. PMID- 10481612 TI - Effects of permanent tracheostomy on respiratory reflexes to lung inflation and capsaicin in sevoflurane-anaesthetized dogs. AB - The aim of this study was to evaluate the effects of permanent tracheostomy on baseline breathing, reflex responses to lung hyperinflation (1.5 kPa) and right atrial capsaicin injection (5 micrograms/kg) before and at 3 and 5 weeks after tracheostomy in sevoflurane-anaesthetized spontaneously breathing dogs (n = 6). In all observation periods, apnoeic responses, represented by an increase in expiration time after lung inflation and right atrial capsaicin injection, were consistently observed to be a result of the Hering-Breuer inflation reflex and the pulmonary C-fibre chemoreflex. Investigation at 3 and 5 weeks after tracheostomy revealed no significant evidence of changing baseline breathing pattern or reflex responses to lung inflation and capsaicin in any ventilatory variable (inspiration time, expiration time, tidal volume, expired ventilation, and end-tidal PCO2) from levels recorded before surgery. These results indicate that permanent tracheostomy, at least up to 5 weeks, does not affect the baseline breathing pattern, the Hering-Breuer inflation reflex or the pulmonary C-fibre chemoreflex. Further, our investigation provides a useful canine model in respiratory physiology. PMID- 10481613 TI - Effect of two dosages of d-cloprostenol on intrauterine pressure and uterine motility during dioestrus in experimental cows. AB - Prostaglandins are commonly used in herd management for their luteolytic properties; however, prostaglandins also have a direct contractile effect on the myometrium. We hypothesized that different dosages (0.15 and 0.3 mg) of d cloprostenol, a synthetical prostaglandin F2 alpha preparation, would differ as to their contractile effects on the uterus. Intrauterine pressure was recorded during dioestrus of lactating dairy cows, using a transcervically placed intraluminal pressure microtransducer. After recording of physiological uterine motility for 30 min, a placebo or d-cloprostenol at one of the two different dosages was administered intramuscularly, followed by a 2-h recording period. Significant differences were found for the area under the curve (P < or = 0.05) and mean amplitude (P < or = 0.05), whereas the number of spikes per 15 min and the baseline pressure during the last 3 min of every 15-min period did not differ significantly among treatments. Peak values for area under the curve and mean amplitude were found between 15 and 30 min after administration of the lower dosage of d-cloprostenol, and between 75 and 90 min after administration of the higher dosage. Using the higher dosage of d-cloprostenol, a steady plateau from 15 to 30 min after administration until the end of the recording session was obtained. Thus, double the luteolytic dose of the d-cloprostenol preparation gives a significantly better reaction in terms of uterine contractility than the single dose. PMID- 10481614 TI - Comparison of cardiopulmonary effects of medetomidine-midazolam, acepromazine butorphanol and midazolam-butorphanol in dogs. AB - Cardiopulmonary effects of intramuscular administration of medetomidine-midazolam at 20 micrograms/kg and 0.3 mg/kg, respectively (MM), acepromazine-butorphanol at 0.05 and 0.2 mg/kg, respectively (AB), and midazolam-butorphanol at 0.1 and 0.2 mg/kg, respectively (MB), were compared in dogs. MM produced relatively large cardiovascular changes such as bradycardia, hypertension, a decrease in cardiac output and vasoconstriction. AB produced moderate cardiovascular changes. Hypotension was observed within 5 min after administration and the mean arterial blood pressure decreased by 30% at 20 min after administration. MB produced decreases in arterial blood pressure and cardiac output, but these changes were small and, of the combinations evaluated in this study, MB had the mildest cardiovascular effects. PMID- 10481615 TI - Pharmacokinetics of indomethacin after intraruminal administration to sheep. AB - The pharmacokinetics of indomethacin (7 mg/kg) was determined in six adult sheep after intraruminal administration. The plasma concentration-time curves showed different patterns for each sheep, depending on the physiological state of the gastrointestinal tract, where absorption took place, and possible enterohepatic and saliva recycling. Indomethacin was irregularly and poorly absorbed from the rumen after intraruminal administration to sheep. Plasma concentrations barely rose to the lower therapeutic margin and mean bioavailability was 38.25 +/- 15.98%. The elimination phase was slow (beta = 0.015 +/- 0.010 h-1) and the half life was long (t1/2 beta = 56.52 +/- 31.21 h). The mean residence time was 20.01 +/- 6.92 h. PMID- 10481616 TI - Cytogenetic variation in six cases of feline mammary tumours. AB - Mammary carcinomas of six feline patients were subjected to karyotypic analysis. A broad variation in the numerical and structural alterations was evident. None the less, several repeatedly occurring alterations were observed. PMID- 10481617 TI - Effects of catecholamines in vitro on lipogenesis in cultured adipose tissue from young goats. AB - Freshly prepared and cultured perirenal and omental adipose tissue explants were used to investigate the effect of age and hormones on lipogenesis in young goats. Perinatal (1-2 days of age) and older (24-32 days of age) male goats were used. Adipose explants were cultured (24 h) in the presence of insulin, cortisol and recombinant bovine growth hormone (bST) and subsequently incubated (2 h) in a glucose-free buffer containing (14C)-acetate in the presence or absence of noradrenaline (Ne) and isoprenaline (Iso) to measure tissue lipogenic responses to hormones added to the culture medium and to measure the responsiveness to catecholamines. Inclusion of hormones in the culture medium did not change lipogenesis during subsequent acute incubation in glucose-free buffer in both perirenal and omental adipose tissue from perinatal goats. On the other hand, in perirenal explants from older animals, insulin alone or insulin plus cortisol increased (P < 0.05) and cortisol alone decreased (P < 0.05) the rate of fatty acid synthesis. When perirenal explants were cultured in the presence of insulin plus cortisol plus bST, the rates of lipogeneses were lower (P < 0.05) than those in cultures with insulin plus cortisol. A similar pattern of the effects of hormones on the rates of fatty acid synthesis was also seen in omental explants; however, these effects were not significant. In vitro rates of lipogeneses were decreased (P < 0.05) by Ne but not Iso in freshly prepared omental explants of both perinatal and older animals and in freshly prepared perirenal explants of older animals. In cultured perirenal explants of both perinatal and older animals the mean values of lipogenesis were decreased (P < 0.05) by both Ne and Iso. However, in cultured omental explants both catecholamines were effective (P < 0.05) in older but not in perinatal animals. PMID- 10481619 TI - Evaluation of selective media for bifidobacteria in poultry and rabbit caecal samples. AB - Five media were evaluated to determine their selectivity for Bifidobacterium sp. in hen and rabbit caecal samples. The colonies arising on the plates inoculated with the caecal samples were Gram stained and screened for the presence of fructose-6-phosphate phosphoketolase activity. Rogosa agar modified by the addition of cysteine-hydrochloride (0.05% w/v), Beeren's agar (with 5 ml/l of propionic acid as a selective agent), BS 2 agar (containing per one litre sodium propionate 15 g, lithium chloride 3 g, paromomycin sulphate 50 mg, neomycin sulphate 200 mg), and Wilkins-Chalgren agar (MW) modified by the addition of acetic acid (1 ml/l) and mupirocin (100 mg/l) were selective for Bifidobacterium sp. from rabbit caecal samples. In contrast, only MW medium was suitable for the isolation and enumeration of bifidobacteria in hen caecal samples. In conclusion, the results suggest that MW agar showed the greatest selectivity. A further advantage of this medium is its case of preparation. Therefore this agar could contribute to the study of the effects of the ingestion both probiotics and prebiotics. Finally, it could be noted that the bifidobacteria selective media should be chosen in respect of the animal species origin of the sample tested. PMID- 10481618 TI - Investigations into the prevention of neonatal Ancylostoma caninum infections in puppies by application of moxidectin to the bitch. AB - The aim of this investigation was to examine whether reactivated Ancylostoma caninum larvae can be eliminated by the administration of moxidectin to pregnant bitches. Four pregnant bitches infected experimentally with 20,000 third-stage larvae of A. caninum were treated subcutaneously with 1 mg moxidectin/kg body weight on day 55 of the pregnancy (5-8 days before parturition). Another four experimentally infected pregnant bitches served as controls. The single moxidectin treatment completely prevented lactogenic infections in the puppies. Neither intestinal stages nor somatic larvae could be found. The administration of moxidectin caused no local or systemic side-effects in the bitches. All 22 puppies of the treated bitches were born healthy and remained so during the whole trial period. Beginning during the third week after birth, all 20 puppies of the untreated bitches developed a severe microcytic, hypochromic anaemia and they revealed a total of 8649 intestinal stages of A. caninum after autopsy. PMID- 10481620 TI - Filamentous intranuclear inclusion bodies in psittacine birds. A structural and ultrastructural study. AB - This paper concerns a disease affecting a group of African grey parrots, which involves intranuclear inclusion bodies composed of filamentous material. The disease was characterized by either sudden death or death within 2-3 days from onset of non-specific symptoms. At necropsy, gross lesions included enlarged liver, mild hepatic congestion and focal necrosis. Samples from five birds were fixed in 10% formol and routinely processed for light and electron microscopy. In four birds, numerous hepatocytes displayed an enlarged nucleus, with peripheral margination of chromatin; the nucleus was partially or wholly filled by a basophilic inclusion body. In the remaining bird, inclusion bodies were acidophilic and completely filled the nucleus; nuclear enlargement was less evident than in the other birds. At ultrastructural examination, and in both types of IIB, nuclei contained looped filaments but no evidence of viral structures. However, virion-like structures were observed in the cytoplasm of some hepatocytes. PMID- 10481621 TI - An outbreak of Newcastle disease in free-living pheasants (Phasianus colchicus). AB - The epidemiology of an outbreak of Newcastle disease in a population of approximately 12,000 free-living pheasants (Phasianus colchicus) on the island of Faeno in Denmark in 1996 is described. The mortality during the epizootic was 56%. The spread of the disease between 7 groups of pheasants could be demonstrated over an observation period of 3 weeks. A total of 70 avian paramyxovirus serotype 1 (APMV-1) isolates was made from the flock. The intra cerebral pathogenicity indices of the 4 isolates tested were in the range 1.78 1.88. By means of immunoperoxidase monolayer assay with murine monoclonal antibodies and sequence analysis of an RT-PCR amplified segment of the F0 viral protein it was found, that the virus belonged to the highly virulent C1 antigenic group and that the amino acid sequence at the F0 cleavage site corresponded with the sequences of virulent APMV-1 strains. Based on the epidemiological circumstances it is believed that the virus was transmitted to the pheasants by feral birds. PMID- 10481622 TI - Epidermal tissue-derived T-cell growth factor, colony stimulating factor and nerve growth factor in chickens. AB - The purpose of this study was to determine the mechanism of the local cytokine mediated immune response in the skin of chickens. The incorporation of 3H thymidine into spleen T lymphocytes from 9-to 10-week-old chickens was augmented by the addition of epidermal tissue culture supernatant (ESN) from 11-day-old embryos. The colony formation of neonatal chicken bone marrow cells in the methylcellulose medium was also significantly increased by addition of ESN. When axonal outgrowth in matrigel was investigated, the embryonal sympathetic ganglion was found to grow axons outwards towards the epidermal tissue specimens. The above results suggest that chicken epidermal cells (probably keratinocytes) produce T-cell growth factor (corresponding to IL-1), colony-stimulating factor for macrophages (M-CSF) and granulocytes (G-CSF), and nerve growth factor (NGF). PMID- 10481623 TI - Outbreak of septicaemic colibacillosis in Japanese quail (Coturnix coturnix japonica). AB - We have carried out an aetiological and pathological study of an outbreak of septicaemia caused by the O165 serogroup of Escherichia coli in Japanese quail (Coturnix coturnix japonica) which resulted in high mortality (90%). After treatment with amoxicilin in drinking water (200 mg/l), morbility and mortality rates dropped markedly. Microbiological studies showed that the organism responsible was an atypical E. coli strain, on the basis of the non-fermentation of lactose (ONPG-), which belonged to the O165 serogroup and was highly virulent for 1-day-old chickens. PMID- 10481624 TI - Lack of immunological cross-reactivity of 36-kDa secretory salivary gland antigen of Hyalomma anatolicum anatolicum with Hyalomma dromedarii and Boophilus microplus ticks. AB - During feeding of ticks of the species Hyalomma anatolicum anatolicum, most of the proteins in salivary gland extracts (SGEs) remained unchanged from the unfed to the fully fed state (from day 1 to day 7 of the experiment), as revealed by SDS-PAGE. However, a 45-kDa protein band disappeared and 26-, 32- and 33-kDa bands appeared when feeding commenced. Some of the protein bands changed their intensity. When probed with anti-H. anatolicum anatolicum hyperimmune sera, transblotted SGE proteins of unfed H. anatolicum anatolicum and Hyalomma dromedarii revealed two common bands of 105 and 80 kDa. A 36-kDa protein band present in H. anatolicum anatolicum SGE could not be detected in H. dromedarii. None of these proteins were detected in partly fed Boophilus microplus when probed with anti-H. anatolicum anatolicum hyperimmune serum. This H. anatolicum anatolicum specific 36-kDa protein was strongly recognized throughout feeding, and thus may be an immunogen of importance for the development of an H. anatolicum anatolicum specific serodiagnostic assay. PMID- 10481625 TI - Intranuclear inclusions in cells infected with Newcastle disease virus. AB - Cells infected by Newcastle Disease Virus were observed to contain both intracytoplasmic and intranuclear inclusion bodies. Ultrastructurally, they consisted of twisted strands of about 18-20 nm diameter resembling nucleocapsids. The presence of these inclusions was detected irrespective of host cell or pathogenicity of the virus. In immunofluorescence and immunogold labelling experiments, these structures were tagged by an anti-P protein monoclonal antibody. In summary, we show that intracytoplasmic and intranuclear inclusion bodies, hitherto used as a taxonomic characteristic for the genus Morbillivirus of the Paramyxoviridae, also occur in a member of the genus Rubulavirus. PMID- 10481627 TI - Protective clothing and heat stress: introduction. PMID- 10481626 TI - Prevalence of verotoxin-producing Escherichia coli (VTEC) in bovine coli mastitis and their antibiotic resistance patterns. AB - Between December 1996 and October 1997, milk samples from a total of 145 cows with coli mastitis were screened for the presence of verotoxin-producing E. coli (VTEC). VTEC were found in four (2.8%) out of the 145 samples. The four isolated strains proved to be verotoxin (VT) 1-, VT2- or VT1- and VT2-positive. However, no strain contained all three virulence factors tested. Further strain characterization was carried out by serotyping as well as by resistance pattern analysis. PMID- 10481628 TI - Heat balance when wearing protective clothing. AB - This issue of the Annals of Occupational Hygiene is dedicated to the topic of heat stress evaluation. For this evaluation, several evaluation programs and international standards are available. In order to understand the reasoning and underlying theory behind these programs and standards, a basic knowledge of heat exchange processes between workers and their environment is needed. This paper provides an overview of the relevant heat exchange processes, and defines the relevant parameters (air and radiant temperature, humidity, wind speed, metabolic heat production and clothing insulation). Further it presents in more detail the relation between clothing material properties and properties of clothing ensembles made from those materials. The effects of clothing design, clothing fit, and clothing air permeability are discussed, and finally an overview of methods for the determination of clothing heat and vapour resistance is given. PMID- 10481629 TI - International standards for the assessment of the risk of thermal strain on clothed workers in hot environments. AB - The International Standards Organisation (ISO) has produced an integrated series of international standards for the assessment of human responses to thermal environments. They include standards for the assessment of thermal comfort, heat stress and cold stress and many have been adopted as European and British standards. This paper describes the series of standards and in particular those concerned with the assessment of risk in hot environments. A three tier approach is taken which involves a simple thermal index that can be used for monitoring and control of hot environments (ISO 7243), a rational approach which involves an analysis of the heat exchange between a worker and his or her environment (ISO 7933) and a standard that describes the principles of physiological measurement which can be used in the establishment of personal monitoring systems of workers exposed to hot environments (ISO 9886). The standards are self-contained and can be used independently. In any comprehensive assessment however they would be used in conjunction. The simple index provides a first stage analysis and can confirm whether or not there is likely to be unacceptable thermal strain. Where a more detailed analysis is required then ISO 7933 provides an analytical method that can provide a more extensive assessment and interpretation leading to recommendations for improvement to the working environment. Where a method needs to be confirmed, or conditions are beyond the scope of ISO 7243 and ISO 7933, then ISO 9886 provides guidance on physiological measurement and interpretation. This would be used in extreme environments where individual responses are required to ensure health and safety or, in the case where personal protective equipment (PPE) is worn, which is beyond the scope of ISO 7243 and ISO 7933. The ISO system therefore covers almost all exposures to hot environments. It would be useful however to extend the scope of the standards that provide a simple index or analytical approach. This paper describes the current standards and their scope and forms the basis and background for descriptions of proposed extensions to the scope of the standards described in other papers in this special issue. PMID- 10481630 TI - Development of a draft British standard: the assessment of heat strain for workers wearing personal protective equipment. AB - Existing methods for estimating heat stress, enshrined in British/International Standards (the Wet Bulb Globe Temperature (WBGT) index [BS EN 27243] and the Required Sweat Rate equation [BS EN 12515; ISO 7933 modified]), assume that the clothing worn by the individual is water vapour permeable; the WBGT index also assumes that the clothing is relatively light. Because most forms of personal protective equipment (PPE) either have a higher insulative value than that assumed or are water vapour impermeable, the Standards cannot be accurately applied to workers wearing PPE. There was, therefore, a need to develop a British Standard which would allow interpretation of these existing Standards for workers wearing PPE. Relevant information was obtained through reviewing the literature and consulting experts. Two questionnaire surveys of potential users of the Standards were conducted, and physiological data collected both experimentally and in work situations were considered. The information collected was used to develop the draft British Standard. It provides information and data on: The general effect of PPE on heat balance of the body (the ability of the body to maintain its 'core' temperature within an acceptable range). The effect of specific forms of PPE on metabolic heat production rate. The thermal insulation and evaporative resistance of types of PPE. The effect of the closure of the garments to the body on heat transfer. The effect of the PPE on the proportion of the body covered. The effect of an air supply (for example, Breathing Apparatus [BA]) to the wearer. Guidance is given on conducting an analysis of the work situation, taking account of the impact of PPE. Detailed methods of interpreting both BS EN 27243 and BS EN 12515 for workers wearing PPE are given, taking account of the factors listed above. Three worked examples using BS EN 27243 and BS EN 12515 are given in the Annex of the draft Standard. PMID- 10481631 TI - Heat stress and protective clothing: an emerging approach from the United States. AB - There is little doubt that heat stress affects many workers adversely and that protective clothing generally adds to the burden. The ACGIH threshold limit value for heat stress is the guiding document for evaluation of heat stress in the United States. Adjustment factors have been used to reflect the change in heat stress imposed by different clothing ensembles. While the first proposed factors started with limited experimental data and professional judgment, heat balance methods in the laboratory have yielded better estimates of adjustment factors and for a wider selection of ensembles. These same experiments have provided the starting point to accounting for nonporous clothing in heat balance evaluation schemes such as required sweat rate. Proposed changes to the ACGIH TLV have been mentioned and a framework for thinking about controls presented. PMID- 10481632 TI - Clothing convective heat exchange--proposal for improved prediction in standards and models. AB - Convection is an important determinant for both sensible and evaporative heat exchange. Heat transfer by convection for normal boundary conditions is readily described by simple power functions. Clothing affects convection in various ways and existing characterisation of clothing by its static insulation values produces inaccurate prediction of sensible heat exchange, eventually leading to erroneous risk assessment. The present paper reviews various methods for evaluation of clothing convective (sensible) heat exchange. Based on available data, two equations are proposed for determination of the reduction of the total insulation values obtained under static, still wind conditions as a consequence of wind and walking effects. The equations apply from 0 to 1.84 clo, from 0.2 to 3 m/s and for walking speeds up to 1.2 m/s. The equations are incorporated in ISO 7933 to provide a more realistic and accurate prediction of sensible heat transfer through clothing. PMID- 10481633 TI - Clothing evaporative heat resistance--proposal for improved representation in standards and models. AB - Clothing heat and vapour resistance are important inputs for standards and models dealing with thermal comfort, heat- and cold-stress. A vast database of static clothing heat resistance values is available, and this was recently expanded with correction equations to account for effects of movement and wind on the static value of heat resistance in order to obtain the dynamic heat resistance of clothing ensembles. For clothing vapour resistance, few data were available so far. Indices for vapour permeability (im) and reduction factors for vapour transfer (Fpcl) of clothing were used instead, using a relation between heat and vapour resistance to derive the clothing vapour resistance from the value for clothing heat resistance. This paper reviews the two commonly used approaches (im and Fpcl), as well as five alternative approaches to the problem. The different approaches were evaluated for their accuracy and their usability. The present paper shows that the currently used relations are not adequate when the wearer of the clothing starts moving, or is exposed to wind. Alternative approaches are shown to improve the determination of dynamic clothing vapour resistance, though some are thought to be too complex. An empirical description of the relation between the clothing permeability index (im) and the changes in clothing heat resistance due to wind and movement was selected as the most promising method for deriving clothing vapour resistance. For this method the user needs to know the static heat resistance, the static im value of the clothing and the wind- and movement-speed of the wearer. This method results in a predicted maximal decrease in clothing vapour resistance by 78%, when clothing heat resistance is reduced by 50%, which is consistent with theoretical expectations and available data. PMID- 10481634 TI - The effects of wind and human movement on the heat and vapour transfer properties of clothing. AB - This paper integrates the research presented in the papers in this special issue of Holmer et al. and Havenith et al. [Holmer, I., Nilsson, H., Havenith, G., Parsons, K. C. (1999) Clothing convective heat exchange: proposal for improved prediction in standards and models. Annals of Occupational Hygiene, in press; Havenith, G., Holmer, I., den Hartog, E. and Parsons, K. C. (1999) Clothing evaporative heat resistance: proposal for improved representation in standards and models. Annals of Occupational Hygiene, in press] to provide a practical suggestion for improving existing clothing models so that they can account for the effects of wind and human movement. The proposed method is presented and described in the form of a BASIC computer program. Analytical methods (for example ISO 7933) for the assessment of the thermal strain caused by human exposure to hot environments require a mathematical quantification of the thermal properties of clothing. These effects are usually considered in terms of 'dry' thermal insulation and vapour resistance. This simple 'model' of clothing can account for the insulation properties of clothing which reduce heat loss (or gain) between the body and the environment and, for example, the resistance to the transfer of evaporated sweat from the skin, which is important for cooling the body in a hot environment. When a clothed person is exposed to wind, however, and when the person is active, there is a potentially significant limitation in the simple model of clothing presented above. Heat and mass transfer can take place between the microclimate (within clothing and next to the skin surface) and the external environment. The method described in this paper 'corrects' static values of clothing properties to provide dynamic values that take account of wind and human movement. It therefore allows a more complete representation of the effects of clothing on the heat strain of workers. PMID- 10481635 TI - The role of performance tests, manikins and test houses in defining clothing characteristics relevant to risk assessment. AB - Clothing is an important determinant of human heat exchange and accordingly a critical factor for heat stress risk assessment. A large number of international standards exist concerning protective properties of clothing. However, few standards deal with ergonomic properties and requirements of clothing, making it difficult to evaluate the function of a clothing ensemble in terms of both protection and physiological strain or discomfort. The paper examines existing test methods and procedures for improvement of the situation. Much of the work are presently at research stages, but should in the near future be available for test houses and consumers. PMID- 10481636 TI - Heat stress and flame protective clothing in mine rescue brigadesmen: inter- and intraindividual variation of strain. AB - A climatic exposure was conducted for the 52 rescue brigadesmen of a mine while they wore flame protective clothing. We looked for individual parameters allowing prediction of tolerated exposure times in the climate tested. Of all individual parameters, only body temperature at the end of the Stoklossa heat tolerance test and physical fitness showed significant influence on the tolerated exposure time, although not very strongly. Age, body mass, and Body Mass Index showed no significant influence on the tolerated exposure time. It was found during a longitudinal study that the tolerance time within the climate for four subjects showed considerable variations, and so it was decided neither to take the result of the heat tolerance test as admittance criterion for the mine rescue service nor to perform a ranking of brigadesmen with respect to heat tolerance by this test. PMID- 10481637 TI - Strategy for evaluation and prevention of risk due to work in thermal environments. AB - A strategy in four successive stages is described and justified for the prevention and control of thermal problems in the workplace. This should allow these problems to be approached and solved progressively in small as well as large companies by relying successively, when necessary, on the complementary competencies of the workers themselves, their technical assistance, the occupational health specialists and the experts. The criteria to fulfil at each stage are described and discussed. Appendix 1 describes in detail the methods to be used at stages 2, "Observation" by the workers and their assistance; at stage 3, "Analysis" with the help of specialists; and outlines the stage 4, "Expertise". PMID- 10481638 TI - Expansion, extension or natural evolution-times they are a changing. PMID- 10481639 TI - Partnership and managed care: a way forward. AB - As the programme of health reform in New Zealand gains momentum the number of new concepts and terms used to describe them is increasing to an extent where it would be easy for nurses, on whom the changes have major impact, to become overwhelmed. Most prominent and influential among the current ideas are managed care, evidence based practice, critical pathways and cultural safety. All these are permeated by the change process. In this paper critical issues surrounding aforementioned terms are discussed from both a professional and a consumer viewpoint, with reference to both the literature and media sources. The purpose is clarification, with the aim of assisting nurses to be more engaged in the debate. PMID- 10481640 TI - Professional closure: constructing the image of New Zealand nursing 1880-1940. AB - The concept of professional closure is a sociological explanation for the way that professions limit their membership. From early in the nineteenth century the medical profession in New Zealand promoted exclusionary closure strategies which ensured their control over the other health-related occupational groups. This paper examines the way that professional closure has shaped the development of nursing in this country since Nightingale nurses arrived in New Zealand in the 1880s, and how professional closure effectively constructed the image of New Zealand nursing through the discourse of vocationism during the sixty year period 1880-1940. PMID- 10481641 TI - Women's infertility: challenges for practice. AB - Infertility is an issue for men and women but it is women who experience the majority of the diagnostic and treatment procedures. Nurses and midwives care for women in a variety of settings in which infertility is managed. This management is currently characterised by an escalation in related technologies and there is the potential for prolonged and complex endeavours to achieve a pregnancy. This paper reviews some pertinent literature from nursing, medicine, feminist scholars and the popular press to consider the ways in which infertility is understood and experienced. The challenge for nurses and midwives is in negotiating the differing perspectives in order to provide appropriate care to each individual woman as she experiences infertility and its management. PMID- 10481642 TI - Cultural allies: creating allegiances across borders. AB - Culture brokerage, as a nursing intervention, is a strategy that is intended to assist nurses to "bridge" the gap between the orthodox health care system and the health belief systems of clients and their families who are from "different" cultures. However, this intervention often reinforces and solidifies borders, rather than expanding or eliminating them. Culture brokerage will be examined against social constructions of "borders", "Others", and the power relationships that influence these constructions. Fein's (1979) "universe of obligation" will provide the framework for a critique of the culture brokerage intervention. This paper argues that culture brokerage maintains existing borders by ignoring the power differentials that exist between the nurse and the client. Reconceptualizing the nurse as cultural ally begins the process of bringing all clients into the universe of obligation. The nurse, as cultural ally, recognizes the differences that exist between the nurse and the client, and asks each to examine how their different lives and experiences are connected. It is through this nurse-client allegiance that borders can be crossed and meaningful relationships created. PMID- 10481643 TI - Agnes Wood--one of the first fifty. PMID- 10481644 TI - Transcultural nursing: new pathways, new ventures. PMID- 10481645 TI - Tomorrow belongs to those who prepare for it today. PMID- 10481646 TI - Caring for the technology dependent child--a case study. AB - Children who are dependent on medical technology present unique challenges for families and health professionals who attempt to care for them at home. The medical and nursing needs of these children challenge and stretch the financial, social and emotional capacities of parents in ways that service providers do not fully understand. Case management is a collaborative parent/nurse model of community nursing practice that has been implemented to meet the needs of children and families with complex special needs where the care goes across primary and secondary care settings. The aim of case management is to facilitate communication and coordination of services between the multi-disciplinary team and the family to achieve desirable goals. This case study describes the special needs of a technology dependent child and the impact of chronic illness on both child and family. From a deeper understanding of the parent's perspective, implications for nursing practice are discussed. PMID- 10481647 TI - Revealing sexuality: have nurses' knowledge and attitudes changed? AB - All nurses should be adequately prepared for assisting clients with issues relating to sexuality. This article describes a descriptive study undertaken between 1988 and 1991 which used a questionnaire to survey the knowledge and attitudes of New Zealand pre- and post-registration nursing students regarding sexuality. The results of this study have previously been available only in an unpublished report. As interest in this area of research is increasing overseas, and as it is now time to consider resurveying New Zealand nurses, it is useful to have a summary of the findings available to a wider audience. Phase One analysed the responses of a convenience sample of 319 registered nurses undertaking a one year post-registration programme in four New Zealand schools of nursing in either 1988 or 1989. Phase Two analysed 575 questionnaires completed by a convenience sample of nursing students in their first and/or third years of a three-year programme leading to nursing registration. Analysis of the 35 true/false items showed that students near the completion of their programme were as knowledgeable or more knowledgeable than registered nurses, although there were areas where both groups lacked information. Analysis of the 33 items measuring attitudes on a 5-point Likert scale suggested that the attitudes of both pre- and post registration students were more liberal than conservative, but with some differences discernible when participants were grouped by demographic variables. Importantly, the study found that 55% of pre-registration students, and 88% of registered nurse participants, felt that nurses were inadequately prepared for helping clients with concerns about sexual matters. The findings are compared with those of studies undertaken overseas this decade. PMID- 10481648 TI - Case management and nurses. AB - Case management, along with calls for review of the position of nurses in healthcare provision, has become increasingly familiar as a feature of the restructuring of health services. There has been considerable confusion about the meaning of case management. Nurses in New Zealand have looked to nurses' experience and research in the United States where the term originated and have undertaken our own research to construct models for our system. This paper provides a sketch of case management developments in New Zealand with reference to American nurses' endeavours. It is an edited version of the discussion paper: "Case Management and Nurses" prepared for the College of Nurses Aotearoa in 1995 1996. It is intended to provide a contemporary reference to prompt discussion of the concept and more extensive analysis and critique of nursing's contribution within the changing health sector. Case management is conceptualised as a feature of health service delivery at the interface between funds management and professional practice. While nurses have played a major part in service delivery initiatives and research, this interface has been addressed only superficially. PMID- 10481649 TI - The transition to registered nurse: the experience of a group of New Zealand degree graduates. AB - Since 1991, entry to the nursing profession in New Zealand has primarily been through a three year degree programme. The purpose of this study was to explore the issues faced by a group of degree graduates in their first year of registered nurse practice and to identify if the degree outcomes (such as critical thinking, problem solving, reflection on practice, research, independent learning and using cultural safety knowledge) had mediated this transition process. Purposive sampling was used to invite five female graduates to take part in two focus groups, one held at 4 months and the other at 9 months after starting work. Qualitative data were collected using semi-structured questions and the taped interviews were analysed for themes. Five themes were identified: accepting responsibility, accepting their level of knowledge, becoming a team member, professional standards, and workplace conditions. Graduates were using their cultural safety knowledge but they perceived their knowledge of research was linked to further academic study. Their ability to critique their own practice was evident but they found it more difficult to challenge their colleagues' practice and the wider agency culture. Implications of the study are discussed in relation to nursing education and preceptor programmes and areas for further research are indicated. PMID- 10481650 TI - Co-authorship--a means to an end. AB - Nursing Praxis in New Zealand (Praxis) seeks to foster publication as a medium for the development of nursing research and scholarship within the New Zealand nursing community. To achieve that vision, this journal has a strong commitment to supporting nurses to write and prepare manuscripts for publication. This article is a continuation of the Praxis Writing for Publication series and it seeks to explore the way in which co-authorship can be an effective tool for assisting authors to publish. As there is so much clinical innovation and critique happening in New Zealand at present, along with the completion of a considerable amount of nursing research, we are surprised that Praxis is not being overwhelmed with manuscripts. Reflecting on our own experiences of completing research, and then having to find the extra energy from somewhere to disseminate the research findings, we appreciate just how daunting the process of writing an article can be. However, writing does not have to be overwhelmingly difficult, or a solitary activity. We believe that co-authorship is one means of overcoming perceived barriers to submitting manuscripts for publication. PMID- 10481651 TI - The Ministerial Taskforce on Nursing. PMID- 10481652 TI - Advanced nursing practice in New Zealand: 1998. AB - In the last decade, change in the health sector has often seemed to deny and thwart the professionalism and expertise of nurses. However, in the first half of 1998, a number of positive initiatives have come to fruition, which support the development of clinical and educational structures for advanced practice nursing in New Zealand. Within one month of each other the Nursing Council published its framework for post-registration nursing education, and the Nurse Executives of New Zealand published a proposal for the development of advanced practice roles: clinical nurse specialist and nurse practitioner. This article will explore these initiatives, and will examine the proposed clinical nurse specialist and nurse practitioner roles in light of experience in the United States in developing advanced or expanded nursing practice. PMID- 10481653 TI - The scope of advanced nursing practice. AB - The paper, a response to the Ministerial Taskforce on Nursing, has been developed to contribute to the current discussion about nursing practice. The evolving conceptualisation of the scope of practice in New Zealand is scanned as the backdrop for critique of the current trend. An overview of some aspects of developments in the United Kingdom and United States serves to highlight some significant issues concerning the potential for nurses to contribute innovatively to health care in the new era of change. An alternative conceptualisation of practice in three nurse role categories is presented as the inter-relationship of scope and expertise to illustrate how the trend to specialisation might be incorporated in a service scheme founded in the discipline of nursing. PMID- 10481654 TI - Establishment and evaluation of a preschool asthma programme: a pilot study. AB - In New Zealand asthma has been recognised as a major health issue for preschool children and their families (Public Health Commission, 1995). While nurses have been involved with Asthma Self management programmes which have improved the outcomes for older children and adults (Garret, Kolbe, Richards, Whitlock & Rea, 1995: Mitchell, Ferguson & Norwood, 1986) self management programmes have rarely been used for preschool children. The aims of the study were to establish a Preschool Asthma Programme and evaluate whether it would enable parents and their children to increase their knowledge, confidence and skills in asthma management, while improving asthma control. Fifteen families with children aged between three and a half and five years with moderate and severe asthma participated; five families in each of the three programmes offered. The programmes were evaluated using both formative and outcome evaluation. Results indicated that the programme increased knowledge and changes in asthma management by parents and children and increased the confidence of parents in both their own ability, and that of their child, to manage asthma. Seven parents and seven children on the programme achieved positive change in all the self management criteria considered and six parents and two children made changes in most of them. Participation in the programme was associated with improved lung function, asthma control and preschool attendance. The programme was more successful if the child was over four years old and had suboptimal asthma control. It is concluded that, contrary to what has previously been thought, preschool children can be taught self management skills. There is evidence that a Preschool Asthma Programme is a useful addition to the conventional management and can result in improvements in asthma management. PMID- 10481655 TI - Scalp cooling: a critical examination. AB - In this paper the author describes the scientific conceptual framework of chemotherapy-induced alopecia. This includes the mechanisms of hair growth, the action of chemotherapeutic agents on the hair, how scalp cooling works and how the procedure is implemented. She critically examines the published literature on scalp cooling efficacy and describes when it should be used and the pros and cons of the treatment. PMID- 10481656 TI - An exploration of the empowerment of nursing students in a clinical context. AB - At Whitireia Community Polytechnic the curriculum philosophy for the Bachelor of Nursing Degree Programme is based on the Treaty of Waitangi which provides a framework to articulate the concepts of Partnership, Governorship, Advocacy and Empowerment. This paper will describe a research study that aimed to explore the phenomenon of empowerment from the perspective of the student nurse. Using grounded theory methodology, and emancipatory and feminist philosophies, six, Year Two nursing student participants were interviewed and asked to define the term empowerment and give exemplars of moments when they had been empowered in a teaching/learning context. Analysis of data led to the discovery of a theory and development of a theoretical model that described the conditions and consequences that led to empowerment or disempowerment for the student. The theoretical model will be presented and explained, and the conditions that enhanced empowerment will be explored. Recommendations from the research included the need for relevant preparation for students prior to clinical placement, preparation of clinicians through preceptor programmes, greater awareness by registered nurses of the communication that is effective in supporting student nurses and support for assisted reflection for students following clinical placements. It was further recommended that the theoretical model be tested in other contexts. PMID- 10481657 TI - Children, too, have spiritual needs. PMID- 10481658 TI - The report of the Ministerial Taskforce on Nursing. PMID- 10481659 TI - Integrating the terminology and titles of nursing practice roles: quality, particularity and levelling. AB - In this paper, written following the recent workshop on advanced practice, the author takes a fresh look at the meaning of expert, specialist and advanced practice. She proposes that they are distinctive and complementary aspects of every nursing role, and suggests a set of attributes for each. PMID- 10481660 TI - The characteristics of traumatic events experienced by nurses on the accident and emergency ward. AB - Nurses who work in high-risk areas, such as emergency wards, are susceptible to traumatic stress. The present study is an exploratory one, which attempts to describe and interpret some event characteristics of traumatic incidents identified by nurses. Nine nurses from the Accident and Emergency wards of two New Zealand hospitals were interviewed using the repertory grid technique. Qualitative analysis was used to compare event characteristics across participants and four underlying themes were identified: personal issues, contextual issues, control issues, or organisational issues. Comparison of these results with previous work with high stress occupations suggests that although event characteristics from all groups can be broadly grouped under four themes, there are some distinct differences in the traumatic impact of some event characteristics for nurses. The results have implications in the development of measures of traumatic stress for nurses and have immediate implications for organisational practice. PMID- 10481661 TI - Developing an evidence base for teaching nursing practice skills in an undergraduate nursing program. AB - There is international concern over inadequate levels of nursing practice skill acquisition in undergraduate programs. Preparation of student nurses in nursing practice skills prior to clinical placements reduces cognitive dissonance, enhances cognitive flexibility and thus facilitates skill acquisition. This research was a contextually modified replication of the work of Alavi, Loh & Reilly (1991) in determining an evidence basis for selecting content for the clinical skills curriculum in an undergraduate programme. Thirty-three senior nurse clinicians from medical/surgical areas in two large hospitals offering student placements were asked to rate the frequency of performance (frequently, occasionally, rarely and never) of 77 skills for the beginning registered nurse. Those skills frequently used and rated over 65% were considered for inclusion in the undergraduate program--yielding 26 nursing practice skills. The clinicians were also asked to list their ten most important skills and the related expected level of competence for the newly registered nurse in order to aid development of a practical skills curriculum for the undergraduate nursing program at Whitireia Community Polytechnic. The research has provided an illuminating glimpse into the world of practice for a newly registered nurse. Future study of the expectations the new graduates have for themselves compared to those of the clinicians would be useful. Further exploration and dialogue between clinicians and faculty could be enhanced through consideration of faculty expectations of the newly graduated nurse and comparison of the three groups as a whole. PMID- 10481662 TI - Decision making the explicit evidence-based way: comparing benefits, harms and costs. AB - Decisions that nurses make in clinical practice are a significant contributor to effectiveness and cost effectiveness of health care. This paper explores the relationship between evidence, decision tools, and the effectiveness of the nursing contribution to health services. Whereas research evidence, to varying extent, has been used to inform decision making in nursing for some time, explicit evidence-based decision-making, or 'balance sheet' decision-making, is less familiar. A continuum of decision-making will be explored within the international trend of the Effectiveness Movement, drawing on international literature and local experience. Included in this discussion will be the concept of explicit evidence-based decision making, the guideline development movement as exemplified through the New Zealand Guidelines Group, information technology in decision-support, and some of the challenges of an evidential approach to nursing. PMID- 10481663 TI - Introducing Louisa Dixon. Interview by Barbara MS Smith. PMID- 10481664 TI - 10th International Conference on Cancer Nursing. PMID- 10481665 TI - Advanced nursing practice. PMID- 10481666 TI - Into the mouths of babes. The government subsidy of infant formula. PMID- 10481667 TI - Cochrane made simple. Anti-D administration during pregnancy for preventing Rhesus isoimmunisation. PMID- 10481668 TI - A mother's experience of Edgware Birth Centre. PMID- 10481669 TI - Why should I do vaginal examinations? PMID- 10481670 TI - It's my first time: be gentle with me. PMID- 10481671 TI - Soya in artificial baby milks. PMID- 10481672 TI - Using women's views to inform and develop quality in midwifery. PMID- 10481673 TI - Team midwives' views on team midwifery. PMID- 10481675 TI - Mentioning the unmentionable. Teaching about unexpected outcomes in antenatal classes. PMID- 10481674 TI - Child sexual abuse and pregnancy. A personal account. PMID- 10481676 TI - Midwife teachers and clinical practice. Part 1: Separated but not divorced. PMID- 10481677 TI - Anti-D. Part 2: Risks and benefits. PMID- 10481678 TI - The smear test. PMID- 10481679 TI - Pleased to meet you. Sandy Kirkman. PMID- 10481680 TI - Which course? Supporting parents when their baby or child dies (Part 1). PMID- 10481681 TI - Taken to the limit. PMID- 10481682 TI - What does a midwife do? PMID- 10481683 TI - Cochrane made simple. External cephalic version for breech presentation at term. PMID- 10481684 TI - It's a student's life. PMID- 10481685 TI - Helping women through crises in their labour. Interview by Jo Hartley. PMID- 10481686 TI - A baby's death. Helping parents make difficult choices. PMID- 10481687 TI - Domestic violence. A midwife's personal account. PMID- 10481688 TI - Screening of pregnant women for HIV: the case against. PMID- 10481690 TI - Birth at home. AB - Recently the National Birthday Trust performed a confidential survey of home births in the United Kingdom. A good response rate was obtained from midwives, who recruited two groups of women prospectively; those planned and accepted as suitable for a home delivery at 37 weeks and a matched group of similar women who were booked for hospital by 37 weeks. Some 16% of such women were transferred to hospital in late pregnancy (4%) or in labour (12%). This figure rose to 40% among the primiparous women in the survey. The survey report presents an analysis of 4,500 home births and 3,300 hospital controls. Outcomes could therefore be presented by the woman's intent or by what actually happened. In essence it seems that a woman who is appropriately selected and screened for a home birth is putting herself and her baby at no greater risk than a mother of a similar low risk profile who is hospital booked and delivered. Home births will probably increase to 4-5% of all maternities in UK during the next decade and this needs preparatory planning. PMID- 10481689 TI - Maternity care of low-risk women: identification of continuing education needs. PMID- 10481691 TI - Midwife teachers and clinical practice. Part 2: What do teachers want from the relationship? PMID- 10481692 TI - When you've got a problem.... PMID- 10481693 TI - Nipple shields. A tool that needs handling with care. PMID- 10481694 TI - Becoming independent. PMID- 10481695 TI - Successful preterm breastfeeding. PMID- 10481696 TI - Heroines. PMID- 10481697 TI - Pleased to meet you. Jean Sutton. Interview by Tricia Anderson. PMID- 10481698 TI - Which course? Understanding caring through the arts. PMID- 10481699 TI - Fit for a home birth. PMID- 10481700 TI - Wasted opportunities. PMID- 10481701 TI - Specialists at the heart of nursing. PMID- 10481702 TI - An individual approach. PMID- 10481703 TI - Social class and nutrition in coronary heart disease. AB - The UK mortality rate for coronary heart disease is higher than other industrialised countries. Inequalities in health mean people at highest risk of CHD are those least likely to be able to adopt healthy diets. Nurses responsible for nutritional health messages need to take into account people's material circumstances. PMID- 10481704 TI - Piloting day-case surgery for prostate resection. AB - Some patients with benign prostatic hyperplasia need surgery. Transurethral vaporisation of the prostate (TUVP) can be managed without an inpatient stay. Specialist nurses have an important role in assessing and following up day-case patients. PMID- 10481705 TI - The search for strategies to reduce nurse wastage. AB - Potential nurse candidates lack realistic information about the profession. The way nurses are interviewed for entry to the pre-registration diploma programme needs restructuring. The bursary system causes problems, especially for mature students with children. PMID- 10481706 TI - The specialist nurse role in Parkinson's disease. AB - The growing elderly population will result in a higher incidence of Parkinson's disease. People with Parkinson's disease experience adverse effects from both the condition and the drugs used to treat it. Nurse specialists can play a key part in improving patients' everyday life and educating other nurses. PMID- 10481707 TI - Renal transplant: long-term effects of immunosuppression. AB - Renal transplant offers patients freedom from dialysis, and can improve quality of life. However, the potential for this improvement can be compromised by the long-term effects of immunosuppression. Nurses have a key role in educating patients about this. PMID- 10481708 TI - A record sheet for pressure sore risk assessment. AB - Pressure sore risk assessment should be used to inform practice. Differences exist in methods of using risk assessment scales. Nurses need a way of recording risk factors quickly and easily in a format which indicates what action should be taken. PMID- 10481709 TI - Developing an effective research question. AB - Ideas for research can come from numerous sources, and should be explored in depth to identify key areas for research. Question stems can be used to generate the research question. The research question must be carefully worded to avoid misinterpretation and ambiguities. PMID- 10481710 TI - Urine reagent strips: an MDA evaluation. AB - Despite being cheap to buy, the annual UK urine reagent strip bill is more than 4 Pounds million. Nurses need to be aware that each company's test strip works differently. They also need to learn about the external factors that can interfere with tests. PMID- 10481711 TI - Dementia. Part 4: Risk management. AB - Negative assumptions that people with dementia are at risk from a variety of dangers often influence the care they receive. Such attitudes are usually based on misunderstandings about the meaning behind observed behaviours. Good risk management practice involves assessing how risk can be managed in ways that enhance autonomy and wellbeing. PMID- 10481712 TI - Nurse prescribing: now and forever. PMID- 10481713 TI - Reducing risk to theatre staff. PMID- 10481714 TI - A proactive alternative to drugs. PMID- 10481715 TI - Nurse prescribing and the Crown review. AB - By March 2001 26,000 district nurses and health visitors will have prescribing authority. Many hospital nurses do not realise the Medicines Act (1968) allows for the dose adjusting and supply of 'homely remedies'. There has been increasing concern about the legality of group protocols, as the term does not exist in law. PMID- 10481716 TI - Manual handling in the operating theatre. AB - Safer transfer practices are needed in operating theatres. An observational study on manual handling in an operating department led to improvements in transfer practice. Staff should have easy access to the appropriate handling aids. PMID- 10481717 TI - The legal framework for informed consent. AB - Touching a patient without consent can leave the nurse open to action for battery. Consent must be informed and not given under duress. Consent can be written, verbal or implied. Patients should be advised if the risk of complications is other than minimal. PMID- 10481718 TI - Using acupuncture and acupressure to treat postoperative emesis. AB - Interest in non-pharmacological strategies to manage health-care problems is increasing. Effective treatment for postoperative nausea and vomiting continues to be elusive, despite the introduction of new anti-emetic drugs. Acupuncture and acupressure might offer an effective, safe, simple and cheap therapy. PMID- 10481719 TI - Hepatic arterial perfusion scintigraphy. AB - Nurses caring for patients with liver cancer may be asked to help administer radiopharmaceuticals and to measure the effectiveness of such treatments. With the right knowledge, they can also help alleviate patient fears. PMID- 10481720 TI - Multidisciplinary case notes: an audit. AB - Patient records must be comprehensive, clear, accurate and updated on a regular basis. A pilot study which introduced multidisciplinary case notes to a mental health-care unit found they fulfilled these requirements. Staff who used the multidisciplinary case notes preferred them to traditional methods of record keeping. PMID- 10481721 TI - Skin care for patients receiving radiotherapy. AB - Radiotherapy can cause significant skin reactions. Research suggests that skin care practices are diverse for patients receiving radiotherapy. Standardised practice enables nurses to evaluate interventions. PMID- 10481722 TI - Analysis of quantitative research data: Part 2. AB - Statistical and clinical significance are different. An observed difference that does not reach statistical significance may still be clinically relevant. Results presented within research reports should always be interpreted with care. PMID- 10481723 TI - Selecting blood glucose monitoring systems. AB - A wide range of blood glucose monitoring systems is available for use in different applications. The systems have varying characteristics which should be taken into consideration during selection. Operator technique can influence the quality of the results. PMID- 10481724 TI - Dementia. Part 3: Communication. AB - People with dementia often have difficulty communicating. Nurses should ensure these difficulties are not due to physical problems such as a lost hearing aid. Taking time to understand them can help nurses to communicate effectively with people with dementia. PMID- 10481725 TI - The name of the game is jargon. PMID- 10481726 TI - Drugs that should not be crushed. PMID- 10481727 TI - Tube feeding: immune-boosting formulas. AB - This first of two articles on enteral nutrition focuses on the formulas that contain special nutrients to enhance immune system function and digestion. We'll review the specific nutrients in enhanced formulas, what researchers have found about their effects on the critically ill, and whether they're worth using. PMID- 10481728 TI - Tube feeding: administration tips. AB - This second of two articles on enteral nutrition discusses the basics of tube feedings, including the types of tubes that are used, when to start a feeding, and some common complications that develop in patients--aspiration and diarrhea chief among them. PMID- 10481729 TI - When nature turns up the heat. AB - Heat-related illness can present as prickly heat, heat cramps, heat exhaustion, or heat stroke. Can you spot the different types and provide the appropriate care? Can you offer advice on how to prevent these disorders in the first place? This article will allow you to answer Yes. PMID- 10481730 TI - Memory lane: a path to compliance. AB - A cantankerous old man had staff members running for cover until they hit upon the key to gaining his cooperation: reminiscence. When Henry went strolling down memory lane, he swept everyone along with him. PMID- 10481731 TI - Got a bright idea? Patent it. PMID- 10481732 TI - A new approach to hemodynamic monitoring. PMID- 10481733 TI - Omega-3 fatty acids: nothing fishy here! PMID- 10481734 TI - If you injure your back on the job. PMID- 10481735 TI - Why has this patient developed sepsis? PMID- 10481736 TI - Molecular characterization of baculovirus Bombyx mori nucleopolyhedrovirus polyhedron mutants. AB - Four newly isolated and two previously isolated polyhedron mutants of Bombyx mori nucleopolyhedrovirus (BmNPV) were studied. Two polyhedron deficient mutants, #126 and #136, produced small uncrystallized particles of polyhedrin in the nuclei and cytoplasm of infected cells. Mutant #211 produced a large number of variably sized polyhedra in the nucleus and #220 produced a few large cuboidal polyhedra in the nucleus. Mutant #24 and #128 were previously isolated BmNPV mutants. Mutant #24 could not produce polyhedrin mRNA and polyhedra produced by mutant #128 lacked oral infectivity. Nucleotide sequence analysis indicated that five mutants (#126, #136, #211, #220 and #128) had amino acid substitutions in polyhedrin and mutant #24 had a point mutation only in the promoter region of the polyhedrin gene. Cotransfection experiments showed that the altered phenotypes were due to the mutations found in the polyhedrin gene regions. In mutants #126 and #136, amino acid sequences of the nuclear localization signal of polyhedrin were identical to those of wild-type BmNPV, suggesting that this sequence was necessary but not sufficient for nuclear localization of polyhedrin. Electron microscopic observation revealed that fewer occluded virions were contained in polyhedra of #128 and #220. PMID- 10481737 TI - Design and construction of African swine fever virus chimeras incorporating foreign viral epitopes. AB - In the present work we have studied the feasibility of introducing foreign epitopes into the African swine fever virus (ASFV) particle by genetic manipulation of the virus. For this purpose, we developed specific transfer vectors containing the gene encoding for the highly antigenic structural ASFV protein p54 in which foreign sequences were introduced. DNA sequences encoding continuous linear epitopes, the antigenic site A from foot-and-mouth disease virus (FMDV) VP1 protein and the DA3 antigenic determinant from transmissible gastroenteritis coronavirus (TGEV) nucleoprotein N, were separately cloned into the p54 gene, in a region encoding a non-essential domain of the protein. Chimeric p54 genes were inserted by homologous recombination into the thymidine kinase (TK) locus of ASFV genome. The resulting recombinant viruses efficiently expressed both chimeric proteins under transcriptional control of the p54 promoter, and the chimeric gene products were recognized by antibodies to both p54 and foreign epitopes. The modified p54 proteins were also found in the viral particles and complemented the function of the wild-type p54, since deletion of the p54 gene from recombinant viruses did not affected virus replication in Vero cells. This work demonstrates for the first time the feasibility of incorporating foreign amino acid sequences (up to 18 residues) into a protein component of the ASFV particle without affecting virus viability. PMID- 10481738 TI - Clinical implications of mutations C-to-T1653 and T-to-C/A/G1753 of hepatitis B virus genotype C genome in chronic liver disease. AB - Among many mutational "hot spots" on hepatitis B virus (HBV) genome, A-to-T1762 and G-to-A1764 within the core promoter have been underscored in view of disease association as well as viral expression/replication. Although to a lesser extent, C-to-T1653 and T-to-V(C/A/G)1753 were also noteworthy in our previous study. To assess the clinical significance of these mutations, we determined the nucleotide sequence of an HBV DNA fragment covering these sites in HBsAg-positive blood donors (n = 160) and patients with chronic hepatitis (n = 66), liver cirrhosis (n = 45), and hepatocellular carcinoma (n = 58), most of whom were infected with genotype C HBV (subtype adr). In cases where HBe antigen was positive, the frequency of T1653 and/or V1753 showed a striking increment from chronic hepatitis patients (18%) to liver cirrhosis and/or hepatoma patients (82%), whereas that of T1762/A1764 was already high in chronic hepatitis patients (76%). In HBe antigen-negative cases, by contrast, significant difference in the frequency of T1653/V1753 mutants was found between blood donors (22%) and chronic hepatitis patients (67%). Our results suggest that T1653/V(particularly C)1753 mutants are more closely associated than T1762/A1764 with the progression of liver disease from chronic hepatitis to cirrhosis in HBe antigen-positive patients. A system of site-directed mutagenesis PCR RFLP was constructed to diagnose T1653 and C/A1753 more conveniently. Detecting T1653 and C/A1753 by this method would contribute to the differential diagnosis of HBV-associated liver disease. PMID- 10481739 TI - Virus-specific cell receptors are necessary, but not sufficient, to confer cell susceptibility to African swine fever virus. AB - The entry of African swine fever (ASF) virus into Vero cells and swine macrophages is mediated by saturable binding sites located in the plasma membrane, which have been related, as in other virus-cell systems, to the sensitivity of the cell to the virus. In order to define this correlation, we have analyzed up to 16 cell lines derived from different species for their sensitivity to virus infection, to determine the step in the virus infective cycle that was blocked in each resistant cell, the presence of saturable cell receptors and the percentage of bound and internalized virus in these cells. Specific ASF virus receptors were found in different quantities in many sensitive and resistant cell lines. The most restricted cells showed a reduced efficiency of virus binding and virus internalization, as well as a lower amount of cell receptors for the virus attachment protein p12. Other resistant cells were restricted only after early virus translation or virus DNA replication, proving that the presence of virus-specific receptors may be necessary, but not sufficient, to guarantee the cell permissiveness to the virus, and that the ASF virus infection can be arrested at different steps on the infective cycle. PMID- 10481740 TI - Differences in the effectiveness of delivery of B- and CTL-epitopes incorporated into the hepatitis B core antigen (HBcAg) c/e1-region. AB - Work from a number of laboratories including our own has shown that foreign B epitopes inserted into the c/e1-region of Hepatitis B core antigen (HBcAg) elicit powerful antibody responses when mice are immunised with the recombinant core particles. In the present study, we wished to take advantage of the immunodominance of the c/e1-region to deliver cytotoxic T-lymphocyte (CTL) epitopes as a recombinant HBcAg vaccine. Our results indicated that recombinant HBcAg containing CTL epitopes of the E7 protein of human papillomavirus failed to prime E7-directed CTL responses when used to immunise mice for antigen processing through either the endogenous pathway via a Salmonella typhimurium vector, or through the exogenous pathway by parenteral immunisation with recombinant core. Hydropathicity plots predict that the presumed surface location of the hydrophilic c/e1-region within the core particle may alter following insertion of hydrophobic residues constituting the CTL epitopes, thereby compromising their presentation to the afferent immune system. Our data indicate that while the c/e1 region has a powerful adjuvanting effect for inserted B-epitopes, it does not serve this function for inserted CTL epitopes. These findings have generic implications for the development of CTL inducing vaccines using HBcAg as a vaccine vehicle. PMID- 10481741 TI - Generation of maize cell lines containing autonomously replicating maize streak virus-based gene vectors. AB - Virion sense gene replacement derivatives of maize streak virus (MSV) were constructed with selectable marker expression cassettes based on the bialaphos resistance gene (bar) and the CaMV 35S promoter. The effect on replication of increasing the genomic size was tested by including: (1) the 550-bp maize adhI intron and 68-bp TMV omega RNA leader sequences upstream of the bar genes; and (2) a fusion between the bar and E. coli glutathione reductase (gor) genes. Three recombinant viral vectors ranging in size from 2.7 kb to 4.8 kb replicated efficiently in biolistically transfected cells of suspension cultured Black Mexican sweetcorn (BMS) cells. Deletions greater than 39 bp 3' of the stemloop sequence in the LIR adversely affected replicon release. Transformed bialaphos resistant BMS cell lines were generated with all three vectors containing the bar gene: between 38 and 60% of cell lines contained replicating viral episomes. The replicons were structurally stable, replicated to copy numbers of over 500 per haploid genome, and were detected for more than one year after introduction. We noted significant enhancement of bar gene expression, both at the protein and RNA levels, associated with the presence of episomal vector DNA. The maize adhI intron and TMV omega RNA leader sequences did not seem to have a significant effect on bar gene expression from replicating constructs, although expression from controls was enhanced. The results suggest that MSV-based constructs would provide a useful system for long-term gene amplification in cereal cell culture systems. PMID- 10481742 TI - Influenza A virus infection modulates the expression of type IV collagenase in epithelial cells. AB - We investigated the effect of influenza A/Beijing/353/89 (H3N2) virus infection on the expression of type IV collagenase in two different types of epithelial cell. Depending on the cell line infected, the viral infection caused changes in the expression of type IV collagenase. The expression of matrix metalloproteinase 9 (MMP-9; 92 kDa) but not of matrix metalloproteinase-2 (MMP-2; 72 kDa) was stimulated in Vero cells. In MDCK cells, the MMP-2 production increased with the virus infection. According to the enzymatic activity revealed with zymography, the MMP-9 promoter activity rose by a factor of over 1788 in influenza A virus infected Vero cells but not in MDCK cells. The tissue inhibitor of metalloproteinase, TIMP-1, had increased slightly (2.3-fold) in Vero cells 48 hours after the infection, but in MDCK cells, influenza A virus had no effect on the TIMP-1 expression. In conclusion, the MMP-9 and -2 expression by influenza A virus infection are modulated at transcriptional level, depending on the epithelial cell line. PMID- 10481743 TI - In vivo and in vitro phosphorylation of rice dwarf phytoreovirus Pns12 cytoplasmic nonstructural protein. AB - In vivo and in vitro phosphorylation and intracellular location of rice dwarf phytoreovirus Pns12, which is encoded by one of the twelve dsRNA genome segments, S12, and comprises 312 amino acids, was investigated. When [32P]phosphoric acid was incorporated into RDV-infected leafhopper cultured cells, labelled Pns12 was immunoprecipitated from the cells by a monospecific anti-Pns12 polyclonal antibody. Recombinant Pns12 was purified from Spodoptera frugiperda cells infected with AcRS12, a baculovirus recombinant carrying a full-length cDNA of RDV S12. Purified Pns12 was also demonstrated to be phosphorylated in vitro by a kinase activity present in host (rice, barley, wheat, leafhopper) cells and non host (tobacco, spinach, white clover, S. frugiperda, mosquito, mammals) cells as well. Immunocytochemical studies showed Pns12 accumulated in the cytoplasm of infected cells, and frequently localized in a slightly electron-dense patch. These results demonstrated that RDV Pns12 was a cytoplasmic nonstructural phosphoprotein. PMID- 10481744 TI - Characterization of unusual G8 rotavirus strains isolated from Egyptian children. AB - We report the first detection of P[14], G8 rotaviruses isolated in Egypt from the stool of children participating in a 3 year study of rotavirus epidemiology. Two strains, EGY1850 and EGY2295, were characterized by a serotyping enzyme immunoassay (EIA), virus neutralization, and sequence analysis of the genes encoding VP7 and the VP8* portion of the VP4 gene. These two strains shared a high level of homology of their VP7s (87.8% nucleotide [nt], 97.2% amino acid [aa]) and VP4s (89.6% nt, 97.1% aa) and had the highest VP7 identity to serotype G8 (> 82% nt, > 92% aa) and VP4 identity to genotype P[14] (> or = 81% nt, > 91% aa) strains. Serological results with a VP7 G8-specific and VP4 P[14]-specific neutralizing monoclonal antibodies supported the genetic classification of EGY1850 and EGY2295 as P[14], G8. Genogroup analysis supports earlier findings that human G8 rotaviruses may be genetically related to bovine rotaviruses. These findings demonstrate that our understanding of the geographic distribution of rotavirus strains is incomplete, emphasize the need to monitor rotavirus serotypes, and extend the known distribution of serotype G8 and genotype P[14] strains in Africa. PMID- 10481745 TI - Overexpression of cellular dihydrofolate reductase abolishes the anticytomegaloviral activity of methotrexate. AB - Cytomegalovirus (CMV) stimulates numerous cellular pathways upon infection. One of these pathways involves activation of dihydrofolate reductase (DHFR), an essential enzyme in the biosynthesis of purines and thymidylate. Here we report that methotrexate (MTX), an inhibitor of DHFR, suppresses murine CMV replication at the level of DNA synthesis in quiescent NIH 3T3 cells. However, MTX has no antiviral activity in NIH 3T3 sublines resistant to MTX due to DHFR overexpression. These results directly link MTX antiviral activity to DHFR and demonstrate that DHFR plays an essential role for CMV replication in quiescent cells. PMID- 10481746 TI - Cloning and expression of yellowtail ascites virus segment A. AB - cDNA of yellowtail ascites virus (YAV) segment A encoding a polyprotein of VP2, NS, and VP3 has been cloned. Comparison of the nucleotide and the deduced amino acid sequences showed very high homology between YAV and other aquatic birnaviruses. The two small open reading frames (VP5) besides the 5' terminus of the VP2 gene were found on segment A of YAV. Proteins encoded by cDNAs from segment A and the serotype-specific epitope region on VP2 were expressed using a baculovirus vector. Western blot analysis confirmed that a polyprotein was expressed and processed into VP2 and VP3 in insect cells infected with the recombinant baculovirus containing the complete polyprotein coding region. In the case of expression in silkworm larvae, only VP3 was detected in hemocytes and fat body of silkworm larvae infected with the recombinant virus. The recombinant fusion protein consisting of VP2 epitope region and polyhedrin was expressed in insect cells and cross-reacted with a mouse monoclonal antibody against VP2 which had a neutralizing activity to YAV. PMID- 10481747 TI - Nucleotide sequence of coat protein gene of yam mild mosaic virus, isolated in Papua New Guinea. AB - We determined the 3'-termimus 1353 nucleotides (nts) in length excluding the poly (A) tail of yam mild mosaic potyvirus (YMMV) RNA. The sequence starts within a long open reading frame (ORF) 1209 nts and is followed by untranslated region (3' UTR) of 144 nts. The coat protein (CP) contains 266 amino acids (aa) with molecular ratio (Mr) of approximately 30 kDa. The CP of YMMV differs substantially from yam mosaic virus (YMV), Japanese yam mosaic virus (JYMV) (57 and 61% of amino acid sequence identity) and other potyvirus species. This result suggests that YMMV should be classified as a new yam potyvirus. PMID- 10481748 TI - Suppression of endothelin system in Vero cells latently infected with influenza virus B/Lee/40. AB - In Vero cells latently infected with influenza virus B/Lee/40 (L/V cells), the endothelin (ET) system was examined as a possible mediator in pathogenesis of latent viral infection by radioimmunoassay (RIA) and quantitative reverse transcription polymerase reaction (RT-PCR). During viral latency of more than two months, ET secretion, preproendothelin-1 (PPET-1) mRNA, and endothelin receptor subtype A (ETAR) mRNA within the cells remained suppressed. Our data indicate that not only the release of ET-1 was downregulated at the transcriptional level but also ETAR mRNA was downregulated rather than upregulated compensatively in L/V cells. PMID- 10481749 TI - Sequence analysis of RNA-2 of different isolates of broad bean wilt virus confirms the existence of two distinct species. AB - The complete nucleotide sequence of RNA-2 from a Japanese isolate IP of broad bean wilt virus (BBWV) was determined. The sequence encodes a single large polyprotein, which contains a putative movement protein and two coat proteins (CPs). The 3'-terminal sequences of RNA-2 were also determined for three other Japanese isolates and two ATCC isolates (PV132 and PV176) of BBWV. The CPs of the four Japanese isolates share 86.8-98.0% amino acid sequences homology with one another and 88.3-96.5% with those reported for the isolate PV131 (BBWV-2). However, they have only 57.9-66.2% homology with those of PV132 and PV176 (BBWV 1). PMID- 10481750 TI - Functional analysis of the heterologous NSP1 genes in the genetic background of simian rotavirus SA11. AB - Function of rotavirus NSP1 was analyzed by using single-NSP1 gene-substitution reassortants, SKF, SDF, and SNF which have the NSP1 gene derived from human rotaviruses KU, DS-1, and canine rotavirus K9, respectively, in the genetic background of simian rotavirus SA11. The NSP1 genes from KU, DS-1, K9, and SA11 exhibited 58-76% nucleotide sequence identity to one another. No substantial difference in viral growth was observed among the reassortants and SA11. However, production of NSP1 was not detected in SNF when viral proteins were labelled with 35S-methionine during replication in MA104 cells, in contrast to SA11, SKF and SDF which exhibited evident expression of NSP1. Difference in reassortant formation was examined among the reassortant clones generated between human rotavirus strain 69M and either of SA11, SKF or SNF. Although reassortant formation rate was significantly lower in the cross 69M x SNF than the other crosses, selection rates of RNA segments from parent strain 69M in the resultant reassortants was similar among the crosses. Selectivity of homologous and heterologous NSP1 genes in SA11 background was also analyzed by mixed infection and multiple passages among the single-NSP1 gene-reassortants and/or SA11, KU NSP1 gene was selected most frequently, whereas homologous (SA11) NSP1 gene was least efficiently segregated. These results indicated that viral growth and genome segment reassortment with other viruses may not be influenced by the presence of heterologous NSP1 and its expression level, while genomic diversity of NSP1 genes might have been associated with the relative adaptability to the genetic background of SA11. PMID- 10481751 TI - Analysis of a translational enhancer present within the 5'-terminal sequence of the genomic RNA of potato virus S. AB - When present as a transcript leader the 5' untranslated sequence from the potato virus S genomic RNA molecule enhances translation of a downstream open reading frame both in vitro and in vivo. Translational enhancement was 30-fold in rabbit reticulocyte lysate and 15 fold in wheat germ above translation from a transcript with a synthetic leader. Transient expression experiments using tobacco protoplasts and particle bombardment of leaf tissue resulted in enhancement of fourteen and five-fold, respectively, above translation with a synthetic leader. In stably transformed plants the PVS 5'UTR enhanced translation yield ca. 5-fold compared with a synthetic 5'UTR. PMID- 10481752 TI - Virus nomenclature. PMID- 10481753 TI - [AIDS--the forgotten pandemic?]. PMID- 10481754 TI - [Neurocardiogenic syncopes in patients with implanted pacemakers]. AB - BACKGROUND AND OBJECTIVE: Recently the tilting table test (tilting up to 70 degrees angle) has established itself for the diagnosis of neurocardiogenic syncope (NS). Usually patients with bradycardic arrhythmias or asystole are treated by pacemaker implantation. But if the syncope is of neurocardiogenic aetiology other therapeutic alternatives must be chosen. We here report on seven patients in whom a pacemaker had been implanted in the treatment of syncopes, in ignorance of their neurocardiogenic aetiology, yet they had recurred. PATIENTS AND METHODS: Pacemakers had been implanted in eight patients: two with sick sinus syndrome, three with sinus bradycardia, one with brady-tachyarrhythmia, one with asystole and one with Mobitz-type 2 degrees AV block. All patients continued to have syncopes, when sitting or standing, months to years after the pacemaker implantation. The tilting table test (up to 30 min at an angle of 70 degrees) was positive in all patients. There was no case of pacemaker malfunction. RESULTS: One patient declined further treatment. The remaining seven patients were symptom free in the tilting table test on various medications: theophylline up to 2 x 350 mg, metoprolol up to 2 x 100 mg or disopyramide 3 x 100 mg. CONCLUSION: Patients with syncope while sitting or standing should undergo the tilting table test to determine whether the cause is neurocardiogenic so that pacemaker implantation can be avoided. The majority of patients with NS can be successfully treated by drugs. But in some patients pacemaker implantation may have to considered if drug treatment has failed. PMID- 10481755 TI - [Thoracic splenosis with symptoms of coronary heart disease]. AB - HISTORY AND REASON FOR ADMISSION: A 53-year-old man, who had undergone a splenectomy for a traumatic splenic tear 20 years previously, was admitted because an acute myocardial infarction was suspected. 20 years ago, a splenectomy had been performed because of traumatic rupture. INVESTIGATIONS: Laboratory values were normal except for a microcytic anaemia, thrombocytopenia (109,000/microliter) and a raised ferritin level (547 ng/ml). Coronary heart disease as well as other cardiac and upper gastrointestinal abnormalities were excluded. Computed tomography revealed a space-occupying mass in the left lower lobe of the lung near the heart. This filled with contrast medium on coelangiography at the same time as the liver and a subphrenic mass thought to be an accessory spleen. TREATMENT AND COURSE: At thoracotomy numerous splenic masses, maximally 7 cm in diameter, were removed from near the heart, the costal pleura and the lung margin. There were haematological changes similar to those after splenectomy: leucocytosis (32,400/microliter) and thrombocytosis (637,000/microliter) on the 11th postoperative day and Howell-Jolly bodies noted on the 16th day. CONCLUSION: In splenosis the disseminated pieces of splenic tissue can partly substitute for the normal spleen's functions. Splenosis tissue should, therefore, not be removed in every case, especially, if it does not cause symptoms. PMID- 10481756 TI - [Giant condylomata acuminata (Buschke-Lowenstein tumor)]. AB - HISTORY AND ADMISSION FINDINGS: A 59-year-old woman had for 3 weeks been suffering from painful, moist skin changes on the external genitals, for 3 months having noticed vegetations in the anogenital and perineal region. She had been fatigued and lacking in energy since then. Her last gynecological examination had been 20 years ago. Examination of the skin on admission revealed extensive plaque and cauliflower-like warts and large areas of maceration in the anogenital region, the labia and perineum, with large areas of maceration. Nodules were clearly palpable in the left labium majora: the inner aspects were markedly reddened and had some cutaneous erosions. These findings suggested giant condyloma (GC; Buschke-Lowenstein tumour). INVESTIGATIONS: Biopsy showed marked condylomatous epithelial proliferations which, with the formation of markedly thickened epithelial cones, had reached the stage of a verrucous carcinoma. TREATMENT AND COURSE: Under local anti-inflammatory and adstringent medications the inflammatory reaction quickly subsided. Vulvectomy was performed to remove the tumour. CONCLUSION: Giant condyloma (Buschke-Lowenstein tumour) is a rare pseudocancereous lesion. If human papilloma viruses of type 16 or 18 is demonstrated in the lesions, malignant degeneration should be considered. Histological examination is essential to differentiate it from squamous cell carcinoma. PMID- 10481757 TI - [Interventional endoscopy in chronic inflammatory bowel disease]. PMID- 10481758 TI - [Clinical aspects and molecular genetics of the long QT-syndrome]. PMID- 10481759 TI - [Additional drug therapy in acute non-varicose upper gastrointestinal bleeding]. PMID- 10481760 TI - [Detection of hepatitis A antibodies falsified through vaccination?]. PMID- 10481761 TI - [Foregoing of lifesaving therapeutic measures in moribund patients]. PMID- 10481762 TI - [Foregoing of lifesaving therapeutic measures in moribund patients]. PMID- 10481763 TI - [Significance of clinical research in Germany]. PMID- 10481764 TI - [Atrial fibrillation: a common problem in clinical practice]. PMID- 10481765 TI - Acne therapy. PMID- 10481766 TI - Relevance of positron emission tomography (PET) in oncology. AB - BACKGROUND: The clinical use of positron emission tomography (PET) for detection and staging of malignant tumors is rapidly increasing. Furthermore, encouraging results for monitoring the effects of radio- and chemotherapy have been reported. METHODS: This review describes the technical principles of PET and the biological characteristics of tracers used in oncological research and patient studies. The results of clinical studies published in peer reviewed journals during the last 5 years are summarized and clinical indications for PET scans in various tumor types are discussed. RESULTS AND CONCLUSIONS: Numerous studies have documented the high diagnostic accuracy of PET studies using the glucose analogue F-18 fluordeoxyglucose (FDG-PET) for detection and staging of malignant tumors. In this field, FDG-PET has been particularly successful in lung cancer, colorectal cancer, malignant lymphoma and melanoma. Furthermore, FDG-PET has often proven to be superior to morphological imaging techniques for differentiation of tumor recurrence from scar tissue. Due to the high glucose utilization of normal gray matter radiolabeled amino-acids like C-11-methionine are superior to FDG for detection and delineation of brain tumors by PET. In the future, more specific markers of tumor cell proliferation and gene expression may allow the application of PET not only for diagnostic imaging also but for non-invasive biological characterization of malignant tumors and early monitoring of therapeutic interventions. PMID- 10481767 TI - [Bone marrow recovery after irradiation of the spine]. AB - BACKGROUND: Imaging of bone marrow by radio labeled antibodies against NCA-95 antigen of human granulocytes offers the possibility to visualize granulopoietic activity. After radiotherapy, a reduced uptake delineates the radiation field. Aim of our investigation was the study of strength and duration of this effect. PATIENTS AND METHODS: We analyzed 80 cases of patients irradiated for different primary or metastatic malignancies including parts of the spine who received a bone marrow scan for re-staging or during follow-up. Activity uptake of vertebrae inside and outside the irradiation portal was evaluated by ROI and the ratio was taken as measure of bone marrow suppression. RESULTS: A depression of granulopoietic bone marrow activity was seen even after a few fractions of 1.8 or 2 Gy (Figure 1). Depending on time since irradiation (Figure 2), we found a bone marrow recovery. In patients without bone metastasis, regeneration could be complete, whereas in patients treated for metastases, it was incomplete in all cases (Figure 3). CONCLUSIONS: For a certain time after irradiation, radioimmunoimaging delineates the irradiation portal by showing depression of granulopoiesis. Later on, it also shows bone marrow regeneration after radiotherapy. This may be helpful in reconstruction of radiation portals or in toxicity estimation during multimodal cancer therapy. PMID- 10481768 TI - Metastatic spinal cord compression. Influence of time between onset of motoric deficits and start of irradiation on therapeutic effect. AB - BACKGROUND: In a retrospective analysis we investigated the prognostic significance of the interval between first appearance of motoric deficits and the beginning of radiation therapy (RT) with regard to posttreatment motoric function. MATERIAL AND METHODS: Data of more than 400 consecutive patients being irradiated at our department between 1994 and 1997 because of vertebral metastases were reviewed. Ninety-six patients fulfilled selection criteria including motoric deficits, no preceding surgical or radiotherapeutic treatment of the spine, minimum total dose of 24 Gy referred to spinal cord, and additional treatment with dexamethasone. Two subgroups with a similar number of patients for better comparability were formed according to the time of developing motoric deficits: 1 to 13 days (49 patients) and > or = 14 days (47 patients). Effect of irradiation on motoric function was evaluated 2 weeks and about 3 months after radiotherapy. Patients with severe deterioration of motoric function within 48 hours before radiation therapy (31 patients) were looked at separately. RESULTS: Two weeks after radiotherapy 42/47 patients (89%) developing motoric deficits > or = 14 days showed improvement of motoric function in comparison to 6/49 patients (12%) of the other group. Deterioration occurred in 1/47 patients (2%) of the first and in 24/49 patients (49%) of the latter group. In case of severe deterioration of motoric function within 48 hours before radiation therapy only 2/31 patients (6%) showed improvement, but 20/31 (65%) deterioration. About 3 months after radiotherapy comparable results were observed. Median survival time was 4 months. CONCLUSION: A slower development of motoric deficits before beginning of radiotherapy means a better therapeutic effect and a more favorable functional outcome after treatment. The prognosis is extraordinarily poor if severe deterioration of motoric function occurs within 48 hours before radiotherapy. PMID- 10481769 TI - Effectiveness of hypofractionated radiotherapy in painful bone metastases. Two prospective studies with 1 x 4 Gy and 4 x 4 Gy. AB - BACKGROUND: Although effectiveness of fractionated radiotherapy for painful bone metastases is well documented, there are unanswered questions regarding the adequacy of low-dose short radiation schedules for long-term pain control which give maximum benefit in patients with a short life expectancy. PATIENTS AND METHODS: Two consecutive non-randomized prospective follow-up studies were performed at a single institution to analyze pain response and duration of response in patients with a variety of primary tumors. Included were only patients with symptomatic nonvertebral bone metastases and without impending pathologic fracture. Forty-five patients received 1 x 4 Gy to 50 different sites (group I) while 86 patients received 4 x 4 Gy to 96 sites (group II). Pain relief to irradiation was evaluated according to patient interviews using a 4-point categorical scale. Follow-up was performed 7 and 90 days after radiotherapy. RESULTS: Pain relief after 4 x 4 Gy was significantly superior to 1 x 4 Gy with pain control rates being 86.5% vs 48% at day 7 (after end of treatment) and 80% vs 55% at day 90, respectively. A subgroup analysis of patients treated with 4 x 4 Gy demonstrated a more favorable outcome for breast cancer patients in comparison to patients with other primaries concerning pain relief (96% vs 81%), pain control after 90 days (93% vs 72%), median time to pain progression (9 vs 3 months), and median overall survival (14 vs 5.5 months). CONCLUSIONS: In this study 4 x 4 Gy proved to be clearly superior to 1 x 4 Gy in relieving pain from symptomatic nonvertebral bone metastases without impending pathologic fracture. Even if radiotherapy with 1 single fraction seems to be applicable in specific cases doses higher than 4 Gy should be chosen. In breast cancer patients pain control seems to be better compared to other primaries. PMID- 10481770 TI - [Combined treatment of localized prostate cancer with HDR-Iridium 192 remote brachytherapy and external beam irradiation]. AB - BACKGROUND: The localized prostate cancer can be treated curatively by radiation therapy. The combined treatment of external beam irradiation and HDR-Iridium 192 remote brachytherapy allows higher radiation doses within the tumor without increasing radiation sequelae. PATIENTS AND METHODS: Patients of our clinic have been treated with this combined procedure since 1991. Between 1991 and 1994 15 patients received 2 x 9 Gy of high-dose-rate brachytherapy, followed by 36 Gy external beam irradiation (group A). Because of the frequent local failures in group A, the reference dose of external beam irradiation was increased to 50.4 Gy after brachytherapy between 1994 and 1996. RESULTS: Seven of 15 patients (47%) in group A developed a local recurrence after a median of 17 (13 to 30) months. In group B (20 patients) local failure occurred in 3 patients (15%) after 11, 16 and 32 months. CONCLUSION: The combined radiation therapy of localized prostate cancer cannot substitute radical prostatectomy completely, but it is a promising alternative in the curative treatment in selected patients. PMID- 10481771 TI - Dexamethasone-induced enhancement of resistance to ionizing radiation and chemotherapeutic agents in human tumor cells. AB - BACKGROUND: Dexamethasone-induced changes in radioresistance have previously been observed by several authors. Here, we examined effects of dexamethasone on resistance to ionizing radiation in 10 additional human cell lines and strains, and on resistance to carboplatin and paclitaxel in 13 fresh tumor samples. MATERIAL AND METHODS: Eight human carcinoma cell lines, a glioblastoma cell line and a strain of normal human diploid fibroblasts were arbitrarily chosen for these in-vitro studies. Effects on radiosensitivity were assessed using a conventional colony formation assay. Effects on resistance to the drugs were investigated prospectively (ATP cell viability assay) using 13 fresh tumor samples from consecutive patients operated for ovarian cancer within the context of a Swiss nation-wide randomized prospective clinical trial (SAKK 45/94). RESULTS: Dexamethasone promoted proliferation of 1 of the cell lines without affecting radiosensitivity, while it completely inhibited proliferation of another cell line (effects on radiosensitivity could thus not be examined). Furthermore, dexamethasone induced enhanced radioresistance in 1 of the 8 carcinoma cell lines examined. In the glioblastoma cell line, there was no effect on growth or radioresistance, nor in the fibroblasts. Treatment with dexamethasone enhanced resistance of the malignant cells to carboplatin in 4 of the 13 fresh tumor samples examined, while no enhancement in resistance to paclitaxel was observed. CONCLUSIONS: In agreement with previous reports, we found that dexamethasone may induce radioresistance in human carcinoma cells. Including the published data from the literature, dexamethasone induced enhancement in radioresistance in 4 of 12 carcinoma cell lines (33%), but not in 3 glioblastoma cell lines, nor in 3 fibroblast strains. Dexamethasone also induced enhanced resistance to carboplatin with a similar probability in fresh samples of ovarian cancer evaluated prospectively (in 4 of 13 samples; 31%). We worry that induction of resistance by corticosteroids given to patients undergoing either radiotherapy or chemotherapy with agents causing DNA damage might be associated with a reduced clinical responsiveness in a significant fraction of patients with a carcinoma. PMID- 10481772 TI - [Prospective double-blind study of prophylaxis of radioxerostomia with Coumarin/Troxerutine in patients with head and neck cancer]. AB - AIM: Prospective, randomized placebo-controlled double-blind study to prove the efficacy of Coumarin/Troxerutine (Venalot Depot) for protection of salivary glands during a head and neck irradiation. PATIENTS AND METHOD: Forty-eight radiotherapy patients (60 Gy) with head and neck cancer were included in this trial. During radiotherapy the salivary glands were located in the core irradiation field. Primary efficacy parameters were sialometry, quantitative salivary gland scintigraphy and clinical evaluation of early effects of radiotherapy (RTOG-score, Table 1). All data were collected at 6 assessments: 1 week pre-radiation (U1), at start (U2), half time (U3) and end (U4) of irradiation, 8 days (U5) and 28 days (U6) after the end of irradiation (Figure 1). RESULTS: Twenty-three patients (11 verum, 12 placebo) completed the study with all assessments. Sialometrically, all patients were severely (half of radiotherapy) or completely (end of radiotherapy) xerostomatic (Figure 2). In a global efficacy measure according to O'Brien combining scintigraphy and RTOG there was a tendency for a higher efficacy of verum compared to placebo (p = 0.068). After start of irradiation therapy, the RTOG-score showed continuously and significantly lower early radiation effects under verum than under placebo (U3 vs U6: p < 0.05, area under curve: p = 0.032; Table 2, Figure 3). The scintigraphically determined excretion fraction was slightly less impaired in the verum group compared to the placebo treatment (p = 0.12. Figure 4). There was no difference in drug safety between placebo and verum for adverse events, changes in the activity of liver enzymes and for global impression of tolerability. CONCLUSIONS: The results give support for an advantageous effect of Venalot Depot in the treatment of radiogenic sialadenitis and mucositis. In even a small number of evaluable patients, early clinical effects of irradiation (RTOG-score) were less pronounced in the active treatment group than in the placebo group, but the sample size was too low to prove statistically also the benefit of coumarin/troxerutine with the scintigraphic method. Sialometry seems not suitable for the assessment of early radiation effects. PMID- 10481773 TI - [Results of radiotherapy for vertebral hemangioma]. AB - PURPOSE: Assessment of treatment results of symptomatic vertebral hemangiomas and review of the literature (Table 3). PATIENTS AND METHODS: Ten patients treated between 1974 to 1997 were retrospectively analyzed. Efficacy of treatment was determined according to improvement of pain and/or neurological symptoms (Table 1). RESULTS: Improvement was achieved in 8 of 10 patients. The initially existing neurological symptoms of 3 patients disappeared completely in 2 cases and improved in 1 case. Acute side effects were slight. Late side effects were not seen. A dose-effect relationship could not be assessed. There was no relapse. CONCLUSION: Radiotherapy with 30 Gy for symptomatic vertebral angioma as primary therapy is indicated. In case of neurological symptoms a radiotherapy after operative therapy is recommendable even if the patient is free of symptoms to prevent progress or relapse. PMID- 10481775 TI - [Multimodality therapy of operable pancreatic cancer]. PMID- 10481774 TI - Craniospinal axis irradiation in children. Treatment in supine position including field verification as a prerequisite for anesthesia without intubation. AB - PURPOSE: For craniospinal axis irradiation in young children sometimes anesthesia is required. In order to minimize risks from the anesthesist's point of view supine position would be preferable to standard prone position. In case of irradiation in supine position verification of the 3-field junction in the cervical region causes problems, because there is no direct visual control. For such situations the clinical application of a new technique is presented. PATIENTS AND METHODS: For treatment planning a modern 3D planning system was necessary. Patient's positioning was done by using a vacuum-form body immobilizer and an integrated head mask. Radiation fields were placed only by table movements being calculated by the planning system in relation to a reference point at the patient's surface. In addition to common verification films specially prepared small films were used for the 3-field junction in the cervical region. These films were placed close to the patient for the whole time of each radiation session being exposed by every radiation field. RESULTS: Two children (age 3 and 5 years, respectively) were irradiated as described. Twenty-eight of those specially prepared films were exposed. Two films (7%) had to be excluded because of inadequate exposure. An overlap of radiation fields was seen on 1 of the 26 remaining films (4%), whereas an unacceptable gap was not found. Acute skin reactions were comparable to those observed in patients being irradiated in standard prone position. CONCLUSION: The presented technique for craniospinal axis irradiation in supine position including field verification was not only precise and reproducable, but also comfortable and safe for the patient. We suggest it as a new option for craniospinal axis irradiation in children. PMID- 10481776 TI - [Organ-preserving treatment of bladder cancer with combined radiochemotherapy: Do patients profit from additional neoadjuvant chemotherapy?]. PMID- 10481777 TI - [Diagnosis of recurrent or persistent nasopharyngeal cancers inspite of radiotherapy: Comparison of F-18-deoxyglucose positron emission tomography with computed tomography]. PMID- 10481778 TI - Clinical, angiographic, and sonographic findings after structured treatment of cerebral vasospasm and their relation to final outcomes. AB - OBJECTIVE: To report the early clinical results, quantitative angiographic and sonographic findings, and final outcome in patients with symptomatic vasospasm who had undergone surgical occlusion of the aneurysm and a structured protocol including aggressive intensive care management, endovascular procedures (EP), and barbiturate coma (BC). RESULTS: Thirty consecutive patients (19 women, 11 men, age: 51 +/- 8 years) underwent 38 EP for the treatment of 81 vascular territories (15 balloon dilatations and 66 papaverine infusions). Overall angiographic vasospasm in the intradural ICA improved significantly from 44.7 +/- 19.8% to 16.5 +/- 16%, in the MCA from 44.2 +/- 14.7% to 14.4 +/- 14%, and in the ACA from 38.7 +/- 18.6% to 13.3 +/- 12%. Mean flow velocities (Vm) in the MCA and ACA decreased significantly from 135 +/- 48 cm/sec to 87 +/- 32 cm/sec and from 110 +/- 36 cm/sec to 84 +/- 30 cm/sec, respectively. No significant Vm improvement in the ICA could be demonstrated. Six patients (20%) developed intractable vasospasm after repeated EP and five patients underwent BC. The correlation coefficient between percentage of angiographic vasospasm and Vm increase was -0.19 (p = NS) for the ICA, 0.2 (p < 0.001) for the MCA, and 0.3 (p < 0.05) for the ACA. Correlation coefficient between percentages of angiographic and sonographic improvement was -0.12 (p = NS) for the ICA, 0.42 (p < 0.001), and 0.1 (p < 0.05) for the ACA. Early clinical improvement after EP was observed in 73% of patients and was significantly associated with favourable outcome (GOS 4-5). Sixteen patients (53%) had a GOS 5, six patients (20%) a GOS 4, six patients (20%) a GOS 3, and two patients (6.6%) died as consequence of devastating vasospasm. CONCLUSIONS: Changes in vessel diameter and increases of Vm during vasospasm correlate weakly. In spite of the fact that significant differences in vessel diameter and Vm were demonstrated after treatment, a moderately good correlation between percentages of angiographic and Vm improvement was observed only in the M1 segments. In our experience, a reduction of mortality and disabilities can be achieved with a maximal structured treatment of vasospasm. Early clinical improvement after endovascular treatment is strongly associated with favourable outcome, nevertheless, cost-benefit and controlled trials are necessary to evaluate these techniques. PMID- 10481779 TI - Plasma and cerebrospinal fluid endogenous digoxin-like immunoreactivity in patients with aneurysmal subarachnoid haemorrhage. AB - Recent evidence indicates the presence of naturally occurring digitalis-like compounds in mammals, collectively known as either digitalis-like (DLF) or ouabain-like (OLF) factors, presumed to be endogenous hormones regulating the biological activity of the NA+/ K(+)-ATPase and its isoforms. This substance has been postulated to enhance renal tubular sodium excretion and to increase peripheral vascular resistance. Digoxin-like immunoreactive substance (DLIS) was observed in plasma of some patients with spontaneous subarachnoid haemorrhage (SSAH). Accumulating evidence suggests the central nervous system as a site of synthesis, but also as a site of hypertensinogenic action of endogenous cardioglycosides. The present study intends to establish the ratio of the DLIS in plasma to that in cerebrospinal fluid (CSF) in patients with SSAH and to investigate possible connection of this substance with development of arterial vasospasm. A prospective analysis of DLIS levels was performed on plasma and CSF samples obtained in 40 patients who had suffered a recent SSAH. DLIS levels were determined by the fluorescence polarisation immuno-assay method immediately after the admission to the Ward, and again seven days later. The comparison of CSF and plasma DLIS levels did not show statistically significant differences between the results--neither for the first (Z = 0.530; P = 0.591) nor for the seventh day after the disease onset (Z = 0.448; P = 0.654). Three possible hypothetical explanations of these results are offered: a) substance determined by digoxin immuno-assay has no essential likeness to digoxin; b) loss of the haemato encephalic barrier integrity enabling free substance exchange between plasma and central nervous system; c) digoxin-like substance production within the central nervous system. Further, comparison of DLIS plasma levels (7th day from onset of SSAH) with angiography results showed that patients with multiple vasospasm had essentially higher plasma DLIS levels compared to patients with no vasospasms (Z = 2.59; P = 0.0097). The amount of extravasated blood, assessed on the basis of cranial CT scanning, was also connected with higher plasma DLIS levels (X2 = 3.29; P = 0.0305). The enhanced arterial narrowing which occurs in SSAH may be in part mediated by increased digitalis-like factor activity. PMID- 10481780 TI - Ischaemic complication following obliteration of unruptured cerebral aneurysms with atherosclerotic or calcified neck. AB - We report three cases of ischaemic complications following direct surgery of unruptured cerebral aneurysms having necks with atherosclerotic or calcified walls. Among 30 patients we treated directly for unruptured aneurysm over the last 4 years, 6 had 8 such aneurysms. Atherosclerotic or calcified neck was a major contributor to postoperative ischaemic sequelae in our recent series of unruptured aneurysms treated surgically, and common technical problems during surgery seemed to have caused ischaemic complications in the 3 patients reported here. In this report, attention is given to ischaemic complications in the treatment of such aneurysms. PMID- 10481781 TI - Deterioration of ocular motor dysfunction after transvenous embolization of dural arteriovenous fistula involving the cavernous sinus. AB - We treated 9 patients with the dural arteriovenous fistula involving the cavernous sinus by transvenous embolization. Two patients experienced deterioration of oculo-motor dysfunction after transvenous embolization. We can speculate about two different kind of causes by which patients symptoms deteriorated according to the result of intrasinus pressure recorded during the embolization [1]: high intrasinus pressure caused by the obliteration of the drainage pathway resulted in cranial nerve palsy in one case [2]; implanted coils directly compressed the cranial nerve in another case. Fortunately our cases recovered, but some kind of preventative measures may be needed in similar cases. PMID- 10481782 TI - Basal lateral subtemporal approach for trigeminal neurinomas: report of an experience with 18 cases. AB - Subtemporal craniotomy centred on the external ear canal was used to surgically treat 18 trigeminal neurinomas. The approach was found to be suitable to deal with either or both the middle fossa and the posterior cranial fossa components of the tumour. The basal extension of the exposure was achieved by resection of the roots of the zygomatic arch, roof of the external ear canal and superior third of the mastoid bone. The temporalis muscle was rotated anteriorly. The direction of the approach to the tumour was the shortest and perpendicular from the surface and avoided any neural or vascular exposure or manipulation. The basal exposure was horizontally wide and significantly low which reduced the operating distance, limited the extent of temporal lobe retraction and provided additional space for manipulation of instruments. The exposure was manoeuvreable with anterior, posterior and medial expansion being possible during or prior to tumour resection. The approach had the advantage of being simple and relatively quick and of its familiarity to general neurosurgeons. The experience with the approach with trigeminal neurinomas and its possible advantages over other available approaches to these lesions are analysed in this report. PMID- 10481783 TI - Brainstem glioma: comparative study of clinico-radiological presentation, pathology and outcome in children and adults. AB - Although the clinical and imaging features and behaviour of brain stem gliomas in children are well documented, similar data are not available, for adults. We have carried out a retrospective study, on 101 consecutive patients (71 children and 30 adults) with a histologically verified brain stem glioma. Duration of symptoms, clinical features, imaging characteristics, histopathology and outcome were specifically compared in children and adults with brain stem glioma. Peak incidence was in the first decade in children and in the third and fourth decades in adults. Mean duration of symptoms before admission was 9.7 months in adults and 3.6 months in children (P < 0.001). There were no significant differences in the clinical features between adults and children. Imaging characteristics revealed no major differences except that diffuse hypodense lesions involving the whole brainstem accounted for 41.2% of the lesions in children and only 11.1% of adults (P < 0.001). A stereotactic biopsy was performed in 92 patients and an open biopsy or partial excision in 9 patients. Histopathological examination showed that the majority of gliomas were diagnosed as grade II astrocytomas in both groups. Survival was significantly shorter in children when compared to adults (P < 0.01). While the tumour grade was a significant factor in predicting survival in adults, in children it did not correlate with outcome. Therefore, determination of the grade of a brain stem glioma may be of prognostic significance in adult patients. PMID- 10481784 TI - Molecular pathogenesis of pituitary adenomas: a review. AB - Modern theory of tumorigenesis suggests that genetic alterations may play a role in the initiation and promotion of pituitary adenomas. Gsp and MEN-1 genes play a role in the initiation event, while p53, ras, Rb and nm23 genes play some role in the progression of the tumor. Gsp gene, that may play an important role in 40% of GH-producing tumor, activation of 10% of non-functioning tumors and 6% of corticotroph adenomas, produces cAMP, which stimulates cyclin D1 and D3 which later produce cdk2 and cdk4 respectively, and stimulates cell progression from G1 to S phase. cAMP also induces ras gene, which inhibits binding of pRb with E2F that is necessary to prevent action of E2F in accelerating cell cycle. MEN-1 gene, although found in some sporadic tumors, is more likely associated with familial adenoma. p53, Ras, Rb, nm23 and c-myc genes play some role in the promotion of tumors especially toward their aggressive variant. p53 gene, which is found in up to 60% of ACTH producing adenomas, through action of p21 inhibits progression of cell cycle from G1 to S phase, by inhibiting the action of cyclin D3 on cdk4. Ras oncogene, in cooperation with c-myc gene, prevents the binding of pRb with E2F, which is necessary for preventing progression cell cycle, resulting in progression of cell cycle from G1 to S phase. Nm23 gene inhibits the action of cyclin B and arrests the cell in G2 phase. Further studies will not only be helpful in understanding the genetic pathogenesis and prognosis of pituitary tumors, but also in developing a novel treatment for patients with pituitary adenomas. PMID- 10481785 TI - Postoperative oblique sagittal MR imaging of microvascular decompression for hemifacial spasm. AB - Pre-operative and postoperative oblique sagittal gradient-echo magnetic resonance (MR) imaging was used to evaluate micro-vascular decompression of the facial nerves in 26 patients with hemifacial spasm. The pre-operative MR images were divided into two groups as follows: 22 images in Group I, clear imaging of a high intensity line and/or spot at the root exit zone (REZ) of the facial nerve; and 4 in Group II, and unreliable image around the REZ. Surgery found that the causative vessel was the vertebral artery (VA) in 9 cases and the anterior inferior cerebellar artery (AICA) or the posterior inferior cerebellar artery (PICA) in 13 cases in Group I, and the AICA or the PICA in the 4 cases in Group II. Postoperative MR imaging showed clear decompression as the high-intensity line and/or spot completely separated from the REZ by a low- and/or iso- intensity area in 9 cases of VA compression repositioned to the petrous dura matter, in 11 cases of PICA or AICA compression treated by shredded Teflon pledgets in Group I and in 3 cases in Group II. Postoperative MR imaging showed an incomplete separation of any high-intensity line and/or spot in the REZ in 2 cases of PICA or AICA compression in Group I and in one in Group II. The outcome was excellent in 22 of 23 cases with clear decompression, and in 1 of 3 cases of unclear decompression. Hemifacial spasm persisted in 3 cases. Oblique sagittal gradient-echo MR imaging is a useful method for postoperative follow-up which can demonstrate changes around the REZ of the facial nerve if hemifacial spasm recurs. PMID- 10481786 TI - Limited (L4-S1, L5-S1) selective dorsal rhizotomy for reducing spasticity in cerebral palsy. AB - Selective posterior rhizotomy is effective for relieving spasticity associated with cerebral palsy. In current techniques dorsal roots from L1/L2 to S1/S2 are selectively divided. With transoperative electromyography (EMG) significant sensory loss has been prevented, but postoperative hypotonia following excessive reduction of the fusimotor drive is still of concern for surgeons and therapists. To decrease the volume of deafferentiated rootlets we proposed a limited selective posterior rhizotomy (LPSR) that limits the extent of the surgery to three (L4-S1) or two (L5-S1) dorsal roots. We present the results of two group of spastic children; group 1 (n = 59, 32 quadriplegic and 27 diplegic) who had a L4 S1 LPSR, and group 2 (n = 12) in whom L5 and S1 were selectively rhizotomized. Posture, passive movilization, range of joint movement, and muscle tone in hip flexors, adductors, leg flexors and plantar flexors were graded according to the method proposed by Sindou and Jeanmonod. In all groups these was a significant reduction of the mentioned parameters (Friedman test p < 0.001) at 6, 12 and 18 months after surgery. The preoperative and postoperative ability to ambulate was classified into five grades. In all groups there was a significant (chi 2 between p < 0.01 and p < 0.001) improvement in the quality of their gait. A third of the patients achieved some form of independent ambulation. Our results suggest that extensive selective deafferentation of the lower limbs is not an absolute requisite for reducing muscle tone or achieving functional improvement in spastic children. PMID- 10481787 TI - Chronic subdural haematomas and Parkinsonian syndromes. AB - We describe three men with parkinsonian syndromes caused or aggravated by chronic subdural haematomas. A 63-year-old man developed tremor at rest, rigidity and bradykinesia one week after he fell and hit his head. A 70-year-old patient suffering from tardive dyskinesia and drug-induced parkinsonism experienced deterioration of his bradykinetic symptoms over two weeks. There was no history of trauma. The third patient, a 82-year-old man with idiopathic Parkinson's disease had a marked increase of his left-sided parkinsonian symptoms. Again, there was no history of trauma. In all three patients chronic subdural haematomas were demonstrated by computed tomography. Evacuation of the chronic subdural haematoma resulted in disappearance respectively improvement of the movement disorder. Diagnostic evaluations appear to be delayed and initial misinterpretations are frequent. The findings of our report and review of the literature point out that a favourable outcome after appropriate surgical treatment is achieved in most instances. PMID- 10481788 TI - Direct stereotactic MRI location in the globus pallidus for chronic stimulation in Parkinson's disease. AB - We evaluated the direct location in the globus pallidus (GP) under stereotactic MRI (sMRI) guidance in five parkinsonians treated with chronic deep brain stimulation (four bilaterally). The sMRI consisted of three orthogonal (horizontal, frontal, sagittal) sets of images obtained with a stereotactic frame and its localiser. The sMRI was coupled with ventriculography to compare the location with the classic indirect method based on commissural landmarks. The target was defined on T2-weighted slices in the anterior part of the medial GP, at the vertex of the nucleus. It was reached via one track with a semi-micro electrode and step by step high frequency stimulation, then replaced by a quadripolar electrode once we located the site enabling the optimal clinical improvement. Stereotactic x-rays localised the final position of the electrode. A company software matched sMRI, ventriculography, and peroperative (perop) x-rays, with reference to the stereotactic location boxes. We analysed the effects of acute (perop) and chronic (six-month follow-up) stimulation of active plots (acplots), i.e. leading to optimal clinical improvement. Three distances with reference to the acplots were measured both on sMRI and ventriculography: the laterality from the median sagittal plane of the third ventricle; the anterior position from the midpoint of the intercommissural line (Icl), and the vertical position with regard to the Icl. We then compared the differences in measurements (n = 64) with the Bland and Altman method. The mean difference was 0.09 mm with 95% of the values between +/- 1 mm, but only the laterality had a statistically significant agreement (all the differences included between +/- two times the standard deviation of the mean). The acplots distances from the dorsal, ventral, and medial boundaries of GP (defined by manual surrounding on frontal and horizontal planes) were measured on sMRI. With one exception, the acplots were all included in the nucleus. The six-month acplots were located dorsally with reference to the perop ones. Clinical benefit at six-months follow-up showed results comparable to the literature. Direct location of GP target based on sMRI seems a simple and reliable method. PMID- 10481789 TI - A microsurgical technique for posterior lumbar interbody fusion: technical note. AB - OBJECTIVE: To describe a microsurgical modification of the Ray Threaded Fusion Cage (TFC) instrumentation technique for achieving lumbar interbody fusion. TECHNIQUE: The lumbar fusion is established by application of two titanium cages through two different short skin incisions, first on one and then on the other side. The spinous processes and the whole interspinous ligament are preserved. Under microscopic control, the lower nerve root and foramen are identified and foraminotomy performed. The dural sac is carefully exposed so that a free and gentle retraction of the nerve root to the midline can be obtained. After removal of the disc material, the retractor is inserted. Special attention should be paid to visualization of the superior nerve root in order to avoid neural injuries. The end plate and disc material are then drilled and tapped for insertion of the appropriate cage. CONCLUSION: The Ray TFC lumbar fusion can be practically and safely performed under microscopic control. PMID- 10481790 TI - Effect of niravoline (RU51599), a kappa-opioid receptor agonist, on tumour-origin brain oedema. AB - Niravoline is a selective kappa-opioid receptor agonist. Its effect on tumour origin brain oedema was investigated in a brain tumour model created by implantation of C6 glioma cells into the brain of Wistar rats. Five weeks after the tumour cell implantation, niravoline was administered i.v. at 1 mg/kg, a total of four times at one-hour intervals. In the control group (i.e., saline or vehicle-treated rats), this brain tumour model was found to result in a statistically significant increase in the water content in selected brain regions except for the frontal region, the tumour-containing region and the region surrounding the tumour-containing region. Administration of niravoline inhibited the increase in the water content in the brain regions remote from the tumour. PMID- 10481791 TI - Multiple intraparenchymal schwannomas in the cerebellum, brainstem, and cervical spinal cord. PMID- 10481792 TI - Ependymoma of the cauda equina. AB - Myxopapillary ependymomas are benign tumours which occasionally metastasize along cerebrospinal fluid pathways. Extraneural metastases of spinal ependymomas, however, are very rare, even more so when situated in the pleura. We report the case of a 67 year old woman presenting with shortness of breath after recurrent myxopapillary ependymomas of the cauda equina. Chest X-ray showed multiple pleural lesions diagnosed as metastases of a myxopapillary ependymoma. The MIB-1 proliferation index was 3.1% for the initial spinal tumour, 14.2% for the first and 11.2% for the second recurrence while 12.0% for the pleural metastasis. PMID- 10481793 TI - Bilateral ganglion cysts: case report and review of the literature. PMID- 10481794 TI - Silent pituitary tuberculoma associated with pituitary adenoma. PMID- 10481795 TI - Transient mutism after brain stem infarction. PMID- 10481796 TI - [Biomechanical aspects of malunited distal radius fracture. A review of the literature]. AB - BACKGROUND: One of the most common injuries in orthopaedics is the distal radius fracture. Malunion may lead to pain, limited motion, and loss of grip strength. Presently there is no consensus concerning the indications for corrective surgery after malunion. Studies concerning the biomechanics of the wrist in cases of malunited radius fracture can be helpful. METHOD: Analysis of the literature of the last ten years. RESULTS: In laboratory studies, malalignment of the radius caused alterations of the distal radioulnar joint (anatomically: reduction of the radioulnar contact area, disruption of the deep portion of the dorsal radioulnar ligament, tightness of the triangular fibrocartilage complex; functionally: limited forearm pronation and supination). Further alterations have been found concerning the carpal kinematics, the force transmission through the wrist, the pressure distribution on the articular surface of the radius and the median nerve. CONCLUSION: Attempts should be made to obtain an anatomic reduction of all acute distal radius fractures. Posttraumatic disability of the wrist following malunion in radius fractures should accordingly be treated by a corrective osteotomy of the radius at the original fracture site. PMID- 10481797 TI - [Radio-radial external fixation for correction of malunited distal radius fracture]. AB - For correction of malunited distal radius fractures, the use of the external fixator in radio-radial application was assessed. The device has been used for one-stage correction with corticocancellous bone blocks as well as with hemicallotasis for correction of dorsal angulation. The technical application is described and the use of the external fixator defined. Fourteen patients were treated with radio-radial application, seven of those were corrected with callus distraction. The mechanical possibilities of the fixator allowed complete correction of the malunion. PMID- 10481798 TI - [Results of corrective osteotomy of malunited extension fractures of the radius at the usual site]. AB - BACKGROUND: There is an increasing interest in the corrective osteotomy for the malunited distal radius fracture. Already in the seventies we developed an operative technique for the correction of the malunited Colles' fracture. The technique is based on a special plate, which facilitates precise restoration of the angulation of the joint surface and the length of the radius. The purpose of our study is to investigate the results of our method and to compare them with results in the literature. PATIENTS AND METHODS: We examined retrospectively the patients with a malunited Colles' fracture corrected by our technique until August 1996. The investigated criteria were range of motion, grip strength, pain relief, and X-ray findings. RESULTS: Until August 1996, we have performed 124 corrective osteotomies for malunited Colles' fractures; 115 were carried out by our technique; 91 of them could be examined. The mean follow-up time was 4.5 years. The range of motion has improved in all planes, especially for flexion. Grip strength increased but still remained below the level of the opposite side. On X-ray, average dorsal tilt of the radiocarpal joint was 24 degrees preoperatively and was reduced to an almost normal palmar tilt of 7 degrees. Ulnar-plus variance was reduced from 6 mm to less than 1 mm. CONCLUSION: Corrective osteotomy of the distal radius is of great benefit and leads to improved hand function and diminished pain. The use of our special plate facilitates this ambitious operation considerably. PMID- 10481799 TI - [Results of therapy after primary conservative management of distal radius fractures in patients over 65 years of age]. AB - BACKGROUND: In the clinical routine of our City Hospital Triemli, Zurich, Switzerland, we prefer to treat old patients with fractures of the distal radius conservatively. It was our aim to examine the long-term effects of this treatment on anatomical position, mobility, strength, and patient satisfaction. PATIENTS AND METHODS: We reviewed the data of 102 consecutive patients over 65 years of age. 95 were treated with a plaster cast and controlled weekly in our outpatient department, finally discharged after 50 days. After an average time of eight months, 79 patients could be controlled clinically and radiologically. RESULTS: Although a significant (p < 0.001) reduction of joint mobility and grip strength was found in all cases, we documented no loss of independence in every-day life. Radiological malalignment of the distal radius with a dorsal or palmar angle of more than 20 degrees in the lateral view or a radio-ulnar angle of less than 15 degrees in the dorsopalmar view was associated with increased complaints and patients dissatisfaction. CONCLUSION: In old-aged patients with distal radius fractures, the indication for surgical treatment should be made very carefully, since certain degrees of radiological malalignment are tolerated well. PMID- 10481800 TI - [Conservative versus surgical therapy of perilunar dislocation and dislocation fractures]. AB - 18 perilunate dislocations and fracture-dislocations were treated at the Trauma Center of the Vienna General Hospital during the period from 1992 to 1995. Only five cases were treated without surgery. 15 of these 18 patients returned for follow-up after an average of 16 months. In ten cases good results were achieved. Radiologically, eight patients were classified as good. Overall better results were seen after surgical treatment. PMID- 10481801 TI - [Treatment outcome after surgical arthrodesis]. AB - Different results are reported following total wrist fusion. We performed 64 total wrist fusions due to posttraumatic or degenerative arthrosis of the wrist including Kienbock's disease. Clinical outcome was evaluated using the DASH questionnaire. The mean DASH-function/symptom score was 45.6 points (0 points representing the best possible results and 100 points representing the worst possible result). In addition, 35 patients of this group were reexamined clinically and radiologically using a modified traditional wrist score. The preliminary results of the traditional wrist score and the DASH-score demonstrated a significant correlation (r = -0.71; p < 0.001) indicating the validity of the DASH-score. In conclusion, this study shows that wrist fusion results in significant pain reduction and improvement of function. However, complete pain relief cannot be expected for the majority of patients. The new AO wrist fusion plate is used in our standard technique of wrist fusion. In a further study, we will investigate the outcome of total wrist fusion in comparison with limited wrist fusion using the DASH-questionnaire and the wrist score. PMID- 10481802 TI - [Initial results of treatment with the new AO wrist joint arthrodesis plate]. AB - Although diagnosis and therapeutic options for post-traumatic, idiopathic, and degenerative wrist arthropathies have improved during recent years, total wrist fusion is still frequently regarded as a popular option of treatment. Fourty-one patients underwent wrist arthrodesis between May 1994 and April 1996 using the recently introduced new AO-plate after earlier wrist trauma or Kienbock's disease. Thirty-five patients were examined. The average follow-up time was 15 months. Hand function and active digital range of motion (ADROM) was assessed clinically, grip strength was measured using a Jamar-Dynamometer and the Dexter Computer-System. Patient's daily activities and general postoperative quality of life were estimated with the new DASH-questionnaire. It can be concluded from our data, that the widely claimed complete pain relief after total wrist fusion does not match reality. Eighty percent of the patients complained about reduction of postoperative quality of life with impaired personal hygiene and functional deficits; however, the majority would undergo the procedure again. Seventy percent of the patients returned to their original occupation. The DASH questionnaire proved to be a very useful and sensitive tool for evaluating the patient's problems at the upper extremity. PMID- 10481803 TI - [Radio-lunar Mannerfelt arthrodesis in rheumatoid arthritis]. AB - PURPOSE: We present data of 27 radiolunate and 23 Mannerfelt arthrodeses in patients with rheumatoid arthritis. METHOD AND CLINICAL MATERIAL: Benefits of surgery, i.e. the reduction of pain, swelling, tenderness, and signs of instability as well as radiographic findings of carpal height, carpal collapse, progression of rheumatoid disease beyond the site of wrist fusion were assessed after a mean time of 44 months. RESULTS: Wrist pain and swelling were better managed by Mannerfelt than by radiolunate arthrodesis. Average grip strength of wrists with radiolunate fusion exceeded the strength developed in wrists with Mannerfelt-fusion. However, grip strength of wrists with Mannerfelt-fusion was greater than in the opposite unfused hand, while this was not found in wrists with radiolunate arthrodesis. Postoperative improvements in hand intensive activities were more frequent in the radiolunate fusion population. CONCLUSION: Advancement of carpal degeneration in radiolunate wrist fusions beyond the fusion site may indicate a rising number of symptomatic wrists with increase of follow up time. Our results have led us to suggest radiolunate fusion for the non dominant hand in patients with slow carpal progression of rheumatoid disease and especially if complete wrist fusion of the opposite--often dominant hand--has already been performed. PMID- 10481804 TI - [Morphological aspects of load bearing of the wrist joint after midcarpal partial arthrodesis]. AB - Midcarpal fusion is a reliable treatment for advanced carpal collapse following scaphoid nonunion or scapholunate dissociation. It remains, however, unclear if the alignment of the fused carpal bones influences the redirection of load towards the lunate. The objective of this study was to assess the actual loading conditions after midcarpal fusion in patients by evaluating the patterns of subchondral bone mineralization in the distal articular surface of the radius. Nine patients, who were treated by midcarpal fusion with complete excision of the scaphoid, were examined after an average of 22 months postoperatively by means of CT osteoabsorptiometry. All patients showed peak mineralization in the lunate fossa of the distal articular surface of the radius. Six patients with correct carpal alignment had one large density maximum in the lunate fossa and none in the scaphoid fossa. Patients with incomplete correction of the radial translocation of the capitate, incomplete excision of the scaphoid, or incomplete correction of the extension position of the lunate, showed a second density maximum in the scaphoid fossa. These findings emphasize that a correct carpal alignment is necessary to achieve a complete unloading of the degeneratively altered scaphoid fossa. PMID- 10481805 TI - [Chronic compartment syndrome of the first dorsal interosseous muscle: 2 case reports]. AB - Chronic exercise-induced compartment syndrome of the first dorsal interosseous muscle of the hand is a rare condition. The presented complaint is a dull muscle pain which can be increased by hyperextension of the index metacarpo-phalangeal joint and by repetitive key grip. Two patients complaining of these symptoms could successfully be treated by simple fasciotomy of the first dorsal interosseous compartment. PMID- 10481806 TI - [Neurofibroma of the palm of the hand in Recklinghausen disease. A case report]. AB - We present a six-year-old boy with a slowly growing tumor in the palm of the left hand. Sensibility and motor function were normal, neurofibromatosis Recklinghausen had been diagnosed previously. Surgical treatment allowed macroscopically complete neurofibroma resection, but there was inflammatory infiltration of the flexor tendon sheaths and untreatable fibromatosis within the median nerve proximally. PMID- 10481807 TI - [Osteoid osteoma of the capitate: diagnosis and therapy of a rare cause for wrist pain. Case report and review of the literature]. AB - Occasionally, bone tumors are the source of wrist pain. This is particularly true of osteoidosteoma, which are easily overlooked or diagnosed very late. We present a case report including clinical and radiological findings as well as MRI, surgical treatment, and a review of the current literature. PMID- 10481808 TI - Hepatic metastasis of gastroenterological cancer: its tumor biology and oncological surgery. AB - Tumor angiogenesis and ability of basement membrane invasion for invasion of cancer cells into the tumor vessels and for circulating in the blood stream for hepatic metastasis are required. The molecular detection of cancer cells in the circulatory system strongly indicates the possibility of hepatic metastasis of a gastroenterological cancer. As an effect of the general anesthesia or the surgical procedure, a large amount of cancer cells in the tumor vessels may easily enter into the circulatory system at once from the primary tumor beds. Oncological surgeons should understand that the operation is not only the best treatment for gastroenterological cancer but also the crucial event of hepatic metastasis for cancer cells. Although further investigations are required to determine the relationship between circulating cancer cells and hepatic metastasis, at present, the most important issue to be addressed is the development of new strategies for the prevention of hepatic metastasis following the tumor resection in gastroenterological cancer. PMID- 10481809 TI - [In vitro antimicrobial activity of carbapenem antibiotics against Pseudomonas aeruginosa, measured using a low-concentration Mueller-Hinton Agar culture medium]. AB - Ohya et al. noted that the antibacterial activity of carbapenem-family antibiotics against Pseudomonas aeruginosa was significantly enhanced through lowering the basic amino acid concentration in the culture medium. They reported that there was a marked difference in antimicrobial activity of panipenem (PAPM) against Pseudomonas aeruginosa between the culture medium with Mueller-Hinton Agar (MHA) diluted with distilled water and the non-diluted culture medium. We used 2,312 strains of fresh Pseudomonas aeruginosa, isolated from clinical materials, to examine the antibacterial activity of PAPM in non-diluted and diluted culture media. For testing the susceptibility, we employed the Showa disc method and agar plate dilution method. In the Showa disc method, the inhibition diameters of the tested microbial strains showed a larger distribution for both 16-fold and 40-fold diluted MHA, compared to the non-diluted MHA. The MIC values in the agar plate dilution method were also smaller in distribution for the 16 fold as well as the 40-fold diluted MHA, compared to the non-diluted culture media. Approximately 90% of the strains showed decreased MIC values, 2-8 times in the 16-fold diluted MHA and 2-16 times in the 40-fold diluted MHA. From the above results, we confirmed that the in vitro antibacterial activity of PAPM against Pseudomonas aeruginosa was enhanced through lowering the MHA concentration. PMID- 10481810 TI - [Estimation of antibacterial activity of various antibiotics against Pseudomonas aeruginosa by score method]. AB - Antibacterial activity of various antibiotics against Pseudomonas aeruginosa isolated from each hospitals depends on the variety or amount of antibiotics used in each hospital. The antibiotic, which is effective to P. aeruginosa in a certain hospital is not always effective to that in other hospital. The excellent antibiotics in antibacterial activity have low MIC and hard to progress in resistance, and such antibiotics may be effective against P. aeruginosa isolated from any hospitals. Therefore we thought that the antibiotic, which was progress to resistance, would show a great difference in MIC among hospitals, and we investigated MIC and difference of MIC of various antibiotics against P. aeruginosa isolated from six hospitals. Furthermore, we converted the data of MICs and difference of MIC among six hospitals into the score, and tried to estimate antibacterial activity of various antibiotics by using those scores. From the results of analysis in this report, we think the antibiotics actually surpass in antibacterial activity may be imipenem, cefozopran, cefsulodin and amikacin. New analytical method proposed in this report will become one of potential methods to estimate antibacterial activity of antibiotics against bacteria isolated from inpatient with bacterial infections. PMID- 10481811 TI - [Antimicrobial activities of cefetamet against clinically isolated strains from community acquired respiratory tract infections. Part III]. AB - Antimicrobial activities of cefetamet (CEMT) against clinically isolated strains from patients with community acquired respiratory tract infections were investigated in comparison with those of other oral beta-lactam antibiotics during the period from January to March, 1998. The results are summarized as follows; 1. CEMT showed strong antimicrobial activities against three major pathogens causing community acquired respiratory tract infections, Streptococcus pyogenes, Streptococcus pneumoniae and Haemophilus influenzae. However, antimicrobial activities of CEMT against penicillins (PCs)-intermediate S. pneumoniae (PISP) and PCs-resistant S. pneumoniae (PRSP) were slightly weaker than those of some of the reference antibiotics. 2. No chronological changes of CEMT-MIC level were observed in the antimicrobial activities against S. pyogenes, H. influenzae, Moraxella subgenus Branhamella catarrhalis or Klebsiella pneumoniae subsp. pneumoniae. In contrast to this, due to the increase of PISP and PRSP strains, resistance to CEMT appears increasing with time. PMID- 10481812 TI - [Clinical and bacteriological effects of cefetamet pivoxil against community acquired respiratory tract infections. Part III]. AB - We investigated clinical and bacteriological effects of cefetamet pivoxil (CEMT PI) on community-acquired respiratory tract infection and obtained the following results: This method of investigation was almost the same to those adopted in 1994 and 1996. 1. 512 cases of respiratory tract infection were treated with CEMT PI under the same protocol at a total of 53 institutions in Tokyo, Kanagawa, Saitama, Chiba and Yamanashi prefectures from January, 1, 1998 over March, 31, 1998. Outpatients accounted for 99.7% of all subjects. Diagnoses given to these patients included pharyngolaryngitis (51.4%), tonsillitis (37.7%), and acute bronchitis (10.1%). 2. For the bacteriological study, a manual detailing the method of collecting specimens, storage and transport was distributed to the above-mentioned institutions. The isolation and identification of suspected causative bacteria, determination of minimum inhibitory concentrations (MICs), and investigation of beta-lactamase production were conducted all together at Section of Studies, Tokyo Clinical Research Center. Suspected causative bacteria were detected in 144 (37.2%) out of 387 cases that were the analytical subjects of the clinical efficacy. The major bacteria identified were 32 strains of Streptococcus pyogenes and 19 strains of Haemophilus influenzae. The clinical efficacy (the ratio of improvement) of CEMT by suspected causative bacterium was 84.4% against CEMT-indicated organisms and 81.2% against CEMT-non-indicated organisms. 3. We investigated clinical efficacy rates (the ratio of "markedly improved" + "improved") by disease. The improvement rate was 78.4% in pharyngolaryngitis, 87.0% in tonsillitis, and 79.5% in acute bronchitis. The clinical efficacy rate was an average of 81.9% in all CEMT-PI indicated diseases. PMID- 10481813 TI - Is the societal perspective in cost-effectiveness analysis useful for decision makers? AB - BACKGROUND: The U.S. Public Health Service's Panel on Cost-Effectiveness in Health and Medicine recommends that cost-effectiveness analyses (CEAs) intended to help allocate health resources in the public interest include a reference case analysis, conducted from the perspective of society as a whole. This perspective requires that an analysis measure all health effects and changes in resource use caused by an intervention. VALUE OF SOCIETAL CEAs: Tensions unavoidably arise among the parties to medical decisions--patients, their families and friends, clinicians, and third-party payers. One common approach to handling these tensions is to ignore some of them, to "solve" them by pretending they do not or should not exist. Patients do this when they demand the best care for themselves without regard to the cost to others, payers when they make coverage decisions that shift costs ot others. But by estimating all gains and losses, calculations that reflect the safety, effectiveness, and side effects of an intervention as well as its costs, societal CEAs can help resolve those tensions and provide the basis for decisions that are fair to all parties, an agenda for negotiating such decisions, and information essential for designing compensation and incentives to support them. MAKING BETTER USE OF SOCIETAL CEAs: Use of the societal perspective asks that all parties be aware of and consider the interests of others. Some process or procedure needs to be developed for presenting CEA information to the parties most likely to be affected by decisions, soliciting their views, and negotiating an acceptable decision. This process could be used by government decision makers or by managed care organizations, professional societies, or payers. PMID- 10481814 TI - The once and future application of cost-effectiveness analysis. AB - BACKGROUND: Cost-effectiveness analysis (CEA) is used by payers to make coverage decisions, by providers to make formulary decisions, and by large purchasers/employers and policymakers to choose health care performance measures. However, it continues to be poorly utilized in the marketplace because of overriding financial imperatives to control costs and a low apparent willingness to pay for quality. There is no obvious relationship between the cost effectiveness of life-saving interventions and their application. Health care decision makers consider financial impact, safety, and effectiveness before cost effectiveness. WHY IS CEA NOT MORE WIDELY APPLIED? Most health care providers have a short-term parochial financial perspective, whereas CEA takes a long-term view that captures all costs, benefits, and hazards, regardless of to whom they accrue. In addition, a history of poor standardization of methods, unrealistic expectations that CEA could answer fundamental ethical and political issues, and society's failure to accept the need for allocating scarce resources more judiciously, have contributed to relatively little use of the method by decision makers. HOW WILL CEA FIND GREATER UTILITY IN THE FUTURE? As decision makers take a longer-term view and understand that CEA can provide a quantitative perspective on important resource allocation decisions, including the distributional consequences of alternative choices, CEA is likely to find greater use. However, it must be embedded within a framework that promotes confidence in the social justice of health care decision making through ongoing dialogue about how the value of health and health care are defined. PMID- 10481815 TI - Pooling research results: benefits and limitations of meta-analysis. AB - BACKGROUND: Meta-analysis, the systematic and quantitative synthesis of evidence, has developed considerably in the 1990s and is emerging as an important methodology in medical decision making. As a research methodology, meta-analysis has benefits and limitations that must be acknowledged in its application. EXAMPLES OF BENEFITS: The benefits of meta-analysis include the ability to improve the power of small or inconclusive studies to answer questions and the ability to identify sources of diversity across various types of studies. Meta analysis may reveal how heterogeneity among populations affects the effectiveness of medical interventions in different settings and in different patients. It can also help detect biases, such as publication bias and "Tower of Babel" bias, as well as deficiencies in the design, conduct, analysis, and interpretation of research. In this way, it can also stimulate improvements in the quality of the data needed to optimize medical care. EXAMPLES OF LIMITATIONS: Meta-analysis cannot improve the quality or reporting of the original studies. Other limitations come from misapplications of the method, such as when study diversity is ignored or mishandled in the analysis or when the variability of patient populations, the quality of the data, and the potential for underlying biases are not addressed. CONCLUSIONS: Meta-analysis has promoted the sense that obtaining evidence is a global enterprise and that complete information needs to be evaluated and synthesized to obtain the most unbiased results. Analyzing sources of bias and diversity is essential to performing, understanding, and using meta analyses in medical care. PMID- 10481816 TI - Assessing "best evidence": issues in grading the quality of studies for systematic reviews. AB - BACKGROUND: Evidence-based medicine, clinical practice guidelines, quality and value of health services, and science-based decision making are becoming mainstays of the health care sector. As part of the evidence-based movement, systematic reviews of the literature on clinical questions are becoming increasingly common. Part of the structured approach to evaluating the literature involves assessing the quality of individual studies included in systematic reviews. REVIEW QUESTIONS: To clarify issues in this area, in 1998 the Agency for Health Care Policy and Research commissioned a small project to determine how its 12 Evidence-based Practice Centers were carrying out this part of their systematic reviews (called evidence reports). The number of potential checklists, scales, and similar tools for grading the methodology or the clinical relevance of individual reports is large; the reliability, the validity, the feasibility, and the utility of these tools are either unmeasured or quite variable. CONCLUSIONS: Numerous methodologic questions await definitive research and answers, but in the meantime teams developing authoritative systematic reviews can take certain steps to ensure that their approaches to grading the quality of articles meet applicable scientific standards. Clinicians, program administrators, and health policymakers can then be confident in the overall strength of the evidence and study conclusions. PMID- 10481817 TI - Evidence-based medicine: is translating evidence into practice a solution to the cost-quality challenges facing medicine? AB - BACKGROUND: Evidence-based medicine (EBM) and practice guidelines have been embraced by increasing numbers of scholars, administrators, and medical journalists as an intellectually attractive solution to the dilemma of improving health care quality while reducing costs. However, certain factors have thus far limited the role that EBM might play in resolving cost-quality trade-offs. FACTORS FOR SUCCESS OF EBM RECOMMENDATIONS AND GUIDELINES: Beyond the quality of the guideline and the evidence base itself, critical factors for success include local clinician involvement, a unified or closed medical staff, protocols that minimize use of clinical judgment and that call for involvement of so-called physician extenders (such as nurse practitioners and physician assistants), and financial incentive. TROUBLESOME ISSUES RELATED TO COST-QUALITY TRADE-OFFS: Rationing presents many dilemmas, but for physicians one critical problem is determining what is the physician's responsibility. Is the physician to be the patient's advocate, or should the physician be the advocate of all patients (the patients' advocate)? How do we get physicians out of potentially conflicted roles? EBM guidelines are needed to help minimize the number of instances physicians are asked to ration care at the bedside. If the public can decide to share and limit resources--presumably based on shared priorities--physicians would have a basis to act as advocates for all patients. CONCLUSIONS: Although EBM alone is not a simple solution to the problems of increasing costs and public expectations, it can be an important source of input and information in relating the value of service and medical technology to public priorities. PMID- 10481818 TI - One size does not fit all: questions to answer before intervening to change physician behavior. AB - BACKGROUND: Many interventions have been conducted to change physician behavior, but there is not much evidence regarding their effectiveness. A list of questions is proposed for those who would attempt such interventions: 1. Does the behavior (or decision making) need to be changed? This implies the next two questions. 1a. Is there a logical, evidence-based argument that one decision alternative is preferable for a particular situation? If the would-be behavior changer cannot make an evidence-based argument for changing behavior, there is little moral authority to intervene. 1b. Is there evidence that physicians are not choosing this decision alternative when they should? Interventions are often prompted by evidence that utilization of an alternative was too high or low, but physicians' decisions are not the only determinants of utilization. 2. What is the problem with the decision making? Common sense suggests that different problems require different solutions. Yet interventions are often pursued in the absence of clear information about the reasons physicians did not exhibit the preferred behavior. 3. How could the decision making best be changed? Finding the cognitive problems that caused "wrong" behavior should directly lead to the design of simple, targeted, effective interventions to change this behavior. The judgment and decision making psychology literature suggests that general instruction in reasoning and probability may improve judgments and decision processes. SUMMARY: Physicians' behavior appears to be resistant to change. Understanding why the behavior should be changed and what caused it may make the process of designing interventions more complicated. The resulting interventions, however, are more likely to be simple and successful. PMID- 10481819 TI - [Sea buckthorn and sea buckthorn oils--recent developments in China and central Asia]. AB - Since several years, sea buckthorn and sea buckthorn oils are booming in China, Russia and Central Asia. The background of this development is described and the different species and varieties of sea buckthorn are mentioned. The two different oils obtained from sea buckthorn (pulp oil and seed oil) and their composition is discussed. The use of sea buckthorn and its oils for medical and pharmaceutical purposes has its roots in ancient Tibet. The various "health claims", particularly by Russian and Chinese medical authorities, could not be discussed here. However, a number of recent papers are quoted here without further comment. PMID- 10481820 TI - C18:1, C18:2 and C18:3 trans and cis fatty acid isomers including conjugated cis delta 9, trans delta 11 linoleic acid (CLA) as well as total fat composition of German human milk lipids. AB - In particular with respect to infant nutrition knowledge of the current contents of trans fatty acids (TFA) and of conjugated linoleic acid (CLA in human milk lipids is of interest. After pre-separation by Ag-TLC 11 trans-C18:1 isomers could be quantified by GC with a mean total content of 2.40 +/- 0.60 wt% in samples from 40 German women. For the positional isomers t4, t5, t6-8, t9, t10, t11, t12, t13, t14, t15 and t16 contents of 0.02, 0.02, 0.21, 0.37, 0.32, 0.68, 0.23, 0.15, 0.18, 0.09 and 0.14 wt% were established, with vaccenic acid being the predominant isomer. Further, small trans-C14:1 and trans-C16:1 contents of 0.08% and 0.15% on average were found. As the trans-C18:1 isomers also the trans C16:1 isomers of human milk lipids could for the first time be baseline-resolved by GC to a great extent. Moreover, besides a mean CLA (c9,t11) content of 0.40 +/ 0.09% further 6 cis/trans isomers of linoleic acid with a total content of 1.07 +/- 0.56% on average (w/o CLA) were determined. Further, 4 trans isomers of alpha linolenic acid could be baseline-resolved exhibiting a total content of 0.11%. Altogether German human milk lipids on average were found to contain 3.81 +/- 0.97% TFA with a range of 2.38-6.03%. Direct connections between the dietary intake of trans-C18:1 isomers and the composition of human milk lipids could be established. The major fatty acids exhibited the following contents (wt%): C4: 0.16, C6: 0.18, C8: 0.06, C10: 0.58, C12: 3.12, C14: 6.43, C16: 25.28, C18: 7.41, C18: 1 (total): 33.67, C18: 2 (total): 10.63 and alpha-C18: 3:0.87. PMID- 10481821 TI - [Chemometric evaluation of in vitro studies of bioavailability of zinc in selected foodstuffs and menus]. AB - Using simulated digestion-juices for extraction of 31 foodstuffs and 4 menues, 7 variables were subjected to measuring various polarographic parameters as well as determining antagonists of zinc absorbtion, and were evaluated by chemometric methods. Correlations between the different variables were determined by factor analysis, and 4 factors were extracted. By means of cluster analysis it was established whether similarities between the 31 foodstuffs and 4 menues are existing. The resulting division into groups of bioavailability (high, medium, small) was compared with references based on in vivo methods. A high agreement was established. The found clusters were characterized by discriminance analysis allowing an arrangement of further foodstuffs into different groups of bioavailability. PMID- 10481822 TI - Analysis of Fe content in daily food rations in principal socio-economic groups of population on the basis of questionnaire and analytical studies. AB - The study aimed at estimation of representative rations for the purpose of laboratory reconstruction of the rations on the basis of questionnaire studies in principal socio-economic groups in the Wielko-polska region. The reconstructed representative rations provided material for analytical studies. Sex of individuals and the four seasons of the year were taken into account. In all studied groups, daily food ratios covered lower proportions of recommended dietary allowances for Fe in females than in males. The food rations covered recommended dietary allowances for Fe only in physical or mental male workers. The improper relations between Fe and energy content in food rations of women at the reproductive age were found to deserve a serious concern. Statistical analysis of the results demonstrated that the suggested way of determining representative rations may serve to evaluate Fe nutrition of pre-school children, children of primary schools, university students, mental and physical workers and of retired workers. PMID- 10481823 TI - Extraction of beta-carotene from orange peels. AB - Extraction of beta-carotene from orange peels and its nutritional evaluation through feeding study were carried out to compare the bioavailability of the extractable beta-carotene with that of synthetic beta-carotene. The highest yield of extractable beta-carotene was recovered using a mixture of acetone and hexane (1:1 v/v) as a solvent in a ratio of 15:1 (solvent/sample) and an extraction time of 15 minutes. Growth of rats fed a diet containing beta-carotene extracted from orange peels was normal and their weight gain during feeding period was real. In addition, bioavailability of the extractable beta-carotene was higher than that of synthetic beta-carotene, as a result of the presence of extra oils in the diet that contained beta-carotene extracted from orange peels. Finally, the economical advantage can be reached by extracting beta-carotene from orange peels. The bioavailability of this extractable beta-carotene was high enough to compete with that of commercial synthetic beta-carotene. So, it can be strongly recommended to produce such product in a large scale. PMID- 10481824 TI - Nitrate and nitrite in vegetables from areas affected by war-time operations in Croatia. AB - To evaluate nitrate and nitrite contents in cabbage and potatoes, the most consumed vegetables in Croatia, and to examine if war-time operations in Croatia affected nitrate and nitrite levels in vegetables, potatoes and cabbage cultivated in the war region (East Slavonia) and out of war regions were analyzed. Data showed that nitrate contents were higher in the samples from the war region (32% and 24% in potatoes and cabbage, respectively) but differences were not statistically significant. The nitrate levels found in potato samples (196 mg NaNO3/kg fresh weight (FW) and cabbage samples (911 mg NaNO3/kg FW) were comparable with levels reported for other countries. The nitrite contents in potatoes (321 micrograms NaNO2/kg FW) and cabbage (173 micrograms NaNO2/kg FW) were lower than the contents reported for most other countries. Further examinations of nitrate and nitrite concentrations in vegetables in Croatia as well as examinations of the influence of war-time operations on accumulation of these toxic contaminants are necessary. PMID- 10481825 TI - Homology and convergence in vertebrate and invertebrate nervous systems. AB - Each year the meeting of the American Neuroscience Society attracts over 20,000 members, reflecting the explosion of interest in this field that has occurred over the past few decades. Researchers from many disciplines are focusing their skills on the investigation of every aspect of nervous systems, and neuroscience now encompasses the entire range of endeavour from the study of the single molecules that make up neural membranes to the non-invasive observation of neural function in animals behaving in their natural environments. Advances over the past three decades in our understanding of nervous systems are impressive and come from a multifaceted approach to the study of both vertebrate and invertebrate animals. An almost unexpected by-product of the parallel investigation of vertebrate and invertebrate nervous systems that is explored in this article is the emergent view of an intricate web of evolutionary homology and convergence exhibited in the structure and function of the nervous systems of these two large, paraphyletic groups of animals. PMID- 10481827 TI - Rodlet cells in the head and trunk kidney of the domestic carp (Cyprinus carpio): enigmatic gland cells or coccidian parasites? AB - Rodlet cells have been found in the head and trunk kidneys of the common carp (Cyprinus carpio L). From an experimental sample of 50 carps of various ages, we detected these cells in only seven fishes, contradicting the hypothesis that they constitute a normal component of the fish epithelia. The rodlet cells have a typical structure: 12-16 microns in diameter, with a basal nucleus various in form, and an encasing layer of fibrillar structure. The cells contain rodlets, composed of elongated, opaque sacs featuring dark rods in the center, which strongly elongate in ripening cells. Remarkable pseudopodia-like extensions from the apical parts of the rodlet cells penetrate into the delicate blood vessels and sinusoids of the organs. The encasing layer at the cell apex then opens to release the rodlets into the bloodstream. No junctions were found between the rodlet cells and neighboring cells. It is suggested that these cells comprise some kind of "symbiosis" between leukocyte, possible granulocyte cells, and the parasitic rodlets. The cells serve the rodlets as an incubation chamber, as well as a means of transportation into the bloodstream after ripening. PMID- 10481826 TI - Social rank, stress, fitness, and life expectancy in wild rabbits. AB - Wild rabbits of the two sexes have separate linear rank orders, which are established and maintained by intensive fights. The social rank of individuals strongly influence their fitness: males and females that gain a high social rank, at least at the outset of their second breeding season, have a much higher lifetime fitness than subordinate individuals. This is because of two separate factors: a much higher fecundity and annual reproductive success and a 50% longer reproductive life span. These results are in contrast to the view in evolutionary biology that current reproduction can be increased only at the expense of future survival and/or fecundity. These concepts entail higher physiological costs in high-ranking mammals, which is not supported by our data: In wild rabbits the physiological costs of social positions are caused predominantly by differential psychosocial stress responses that are much lower in high-ranking than in low ranking individuals. PMID- 10481828 TI - The mysterious Mauritian red nectar: a selective toxicant? AB - Using knowledge-based computational structure-activity relationship models, it is hypothesized that the aurone responsible for the uniquely red nectar of several Mauritian bird-pollinated plants functions as a repellant of nectar-robbing or herbivorous mammalian species. PMID- 10481829 TI - Caveat emptor: researcher beware. PMID- 10481830 TI - Hippocampal remodeling and damage by corticosteroids: implications for mood disorders. AB - Mood disorders are common, recurrent and disabling illnesses which are frequently associated with hypothalamic-pituitary-adrenal (HPA) axis dysregulation and memory loss. The hippocampus provides negative feedback to the HPA axis and has an important role in key aspects of spatial and declarative memory. Thus, hippocampal dysfunction could account for both the memory impairment and neuroendocrine abnormalities found in mood disorders. The critical role of the hippocampus in declarative memory, emotional processing, and vulnerability to stress has been demonstrated in both animal and human studies. Cellular processes in the hippocampus including long-term potentiation, neurogenesis, and dendritic remodeling are currently areas of intense study. Human studies report cognitive impairment consistent with hippocampal dysfunction in depression, bipolar disorder, Cushing's disease, and in those individuals receiving exogenous corticosteroids. This review examines data on the role of corticosteroids in hippocampal remodeling and atrophy in patients with mood disorders. Interventions to prevent or reverse the damaging effects of corticosteroids on the hippocampus are discussed. PMID- 10481831 TI - Dose response of adrenocorticotropin and cortisol to the CCK-B agonist pentagastrin. AB - Cholecystokinin (CCK) is an abundant neurotransmitter in brain. Its functional significance in humans is incompletely understood, but it may modulate activity in the hypothalamic-pituitary-adrenal (HPA) axis. To explore this hypothesis, we examined the effects of varying doses (0 to 0.8 microgram/kg) of the CCK-B agonist pentagastrin on adrenocorticotropin (ACTH) and cortisol release in healthy human subjects. We also examined anxiety, heart rate (HR), and blood pressure (BP) responses. Pentagastrin induced large (up to 520% increase over baseline), significant and very rapid, dose-dependent elevations in ACTH and cortisol levels. Significant elevations in HR and BP were seen at all doses, without clear dose-response relationships. Anxious distress and symptom responses were also somewhat dose dependent; but hormonal responses were more robustly linked to pentagastrin dose than to these subjective measures. The HPA axis response to the CCK-B agonist pentagastrin may be a direct pharmacological effect. Further work is needed to determine the mechanisms and the physiological significance of CCK-mediated modulation of the human neuroendocrine stress axis. PMID- 10481833 TI - Altered dopamine activity after recovery from restricting-type anorexia nervosa. AB - When ill, women with eating disorders have disturbances of mood and behavior and alterations of catecholamine activity. It is not known whether these alterations are cause or consequence of pathological eating behaviors. To avoid confounding effects of pathologic eating behavior, we studied women who were recovered (> 1 year, normal weight, regular menstrual cycles, no restricting eating pattern, no bingeing or purging) from anorexia nervosa (AN) and bulimia nervosa (BN) compared to healthy control women. Recovered AN women had significantly lower height adjusted weight than did recovered BN women. CSF HVA (pmol/ml +/- SD), a major metabolite of dopamine, was significantly lower (p < .02) in six restricting-type AN women (131 +/- 49) compared to 19 BN women (216 +/- 73) and at a trend (p < .08) less than 13 bulimic-type AN women (209 +/- 53, p < .06) and 18 control women (202 +/- 57, p < .08). These four groups had similar values for CSF MHPG, a norepinephrine metabolite. Dopamine neuronal function has been associated with motor activity, reward, and novelty seeking. These behaviors are altered in restricting-type AN compared to other eating disorder subtypes. A trait-related disturbance of dopamine metabolism may contribute to a vulnerability to develop this sub-type of eating disorder. PMID- 10481832 TI - Clonidine, but not guanfacine, impairs choice reaction time performance in young healthy volunteers. AB - The present study compares the effects of two alpha 2-adrenoceptor agonists, clonidine (0.5, 2, and 5 micrograms/kg, p.o.), and guanfacine (7 and 29 micrograms/kg, p.o.), in young healthy volunteers on attentional performance. A placebo-controlled double-blind cross-over design (one drug dose/group) was employed. Neither of the drugs affected measures of motor performance or performance at easy levels in an attentional test. However, at the most difficult level in the attentional test, the highest dose of clonidine (5 micrograms/kg), but not guanfacine, decreased the number of correct responses and increased reaction latency. Clonidine 5 and guanfacine 29 micrograms/kg equally increased subjective feelings of sedation and reduced systolic and diastolic blood pressures. Thus, the effects of the drugs on attentional performance could be dissociated from their sedative effects. The results demonstrate that clonidine, but not guanfacine, disrupts performance in an attentional task requiring effortful processing, while leaving performance intact in tests requiring more automatic processing. The lower alpha 2A-vs. alpha 2C-adrenoceptor selectivity ratio of clonidine and the affinity for alpha 1-adrenoceptors of clonidine may be responsible for the different action of these drugs on attention. PMID- 10481834 TI - Anxiogenic-like action of galanin after intra-amygdala administration in the rat. AB - The neuropeptide galanin is expressed in brain structures implicated in regulation of emotionality. The amygdala is known to play a central role in mechanisms of fear and anxiety. We therefore examined the effects of galanin (0.2 and 0.6 nmol/side) on experimental anxiety upon microinjection into the amygdala. Two established animal models of anxiety were used: a punished drinking test, and the elevated plus-maze. Punished responding was dose dependently suppressed by intra-amygdala galanin, whereas unpunished responding, drinking motivation, locomotor activity, and shock thresholds were unaffected. No effects on experimental anxiety were seen in the plus-maze following galanin injection. When injected into parietal cortex, no anxiety promoting properties of galanin were detected. These data suggest that activation of galanin receptors in amygdala modulates neurotransmission involved in fear and experimental anxiety. PMID- 10481835 TI - Platelet peripheral-type benzodiazepine in pregnancy and lactation. AB - The aim of the present study was to investigate the impact of hormonal changes during pregnancy and lactation on the expression of peripheral-type benzodiazepine receptors in platelet membranes. Platelet peripheral benzodiazepine receptor binding characteristics, Hamilton anxiety and depression rating Scores, and progesterone and prolactin (PRL) levels were evaluated during pregnancy and lactation in 17 pregnant women [first (n = 9) and third (n = 8) trimesters], 10 lactating women, and 8 nonpregnant women. A significant decrease (38-41%) in peripheral benzodiazepine receptor density was observed in women during the third trimester of pregnancy when compared to nonpregnant controls and women in their first trimester of pregnancy. The decrease is peripheral benzodiazepine receptors was parallel to the peak in progesterone and PRL secretion. The reduction in peripheral benzodiazepine receptor expression is hormone-dependent and may play a regulatory role geared to prevent pregnancy related overactivity of the hypothalamic-pituitary-ovarian, hypothalamic pituitary-adrenal, and hypothalamic-PRL axes. PMID- 10481836 TI - Inhibition of the high affinity myo-inositol transport system: a common mechanism of action of antibipolar drugs? AB - The mechanism of action of antibipolar drugs like lithium, carbamazepine, and valproate that are used in the treatment of manic-depressive illness, is unknown. Lithium is believed to act through uncompetitive inhibition of inositolmonophosphatase, which results in a depletion of neural cells of inositol and a concomitant modulation of phosphoinositol signaling. Here, we show that lithium ions, carbamazepine, and valproate, but not the tricyclic antidepressant amitriptyline, inhibit at therapeutically relevant concentrations and with a time course similar to their clinical actions the high affinity myo-inositol transport in astrocyte-like cells and downregulate the level of the respective mRNA. Inhibition of inositol uptake could thus represent an additional pathway for inositol depletion, which might be relevant in the mechanism of action of all three antibipolar drugs. PMID- 10481837 TI - 5-HT1B receptors: a novel target for lithium. Possible involvement in mood disorders. AB - Lithium ion is widely used to treat depressive patients, often as an initial helper for antidepressant drugs or as a mood stabilizer; however, the toxicity of the drug raises serious problems, because the toxic doses of lithium are quite close to the therapeutic ones. Thus, precise characterization of the target(s) involved in the therapeutic activity of lithium is of importance. The present work, carried out at molecular, cellular, and in vivo levels, demonstrates that 5 HT1B receptor constitutes a molecular target for lithium. Several reasons suggest that this interaction is more likely related to the therapeutic properties of lithium than to its undesirable effects. First, the observed biochemical and functional interaction occurs at concentrations that precisely correspond to effective therapeutic doses of lithium. Second, 5-HT1B receptors are well characterized as controlling the activity of the serotonergic system, which is known to be involved in affective disorders and the mechanism of action of various antidepressants. These findings represent progress in our knowledge of the mechanism of action of lithium that may facilitate clinical use of the ion and also open new directions in the research of antidepressant therapies. PMID- 10481838 TI - Haloperidol improves membrane phospholipid abnormalities in temporal lobes of schizophrenic patients. AB - Using 31P magnetic resonance spectroscopy, we examined changes in the levels of phosphorus metabolites in the temporal lobes of 13 schizophrenic patients before and 12 weeks after initiating haloperidol treatment. Spectra were obtained from a volume of interest positioned in each temporal lobe. Findings were compared with those in 13 age- and gender-matched healthy subjects. Prior to treatment the patients showed higher levels of phosphodiesters (PDE) in both temporal lobes than healthy subjects. Haloperidol administration significantly reduced the excess of PDE in the left temporal lobe, although the PDE concentration remained somewhat higher bilaterally than in controls. Treatment was associated with a decline in the total symptom score according to the Brief Psychiatric Rating Scale and the score for positive symptoms showed a relatively high correlation with reduction in PDE level in the left temporal lobe. These preliminary results suggest that haloperidol may partially normalize disturbed metabolism or abnormalities in components of membrane phospholipids in the left temporal lobe of untreated schizophrenic patients, paralleling symptom alleviation. PMID- 10481839 TI - Lack of cross-sensitization of the locomotor effects of morphine in amphetamine treated rats. AB - Repeated exposure to morphine and amphetamine induces long-lasting sensitization of their psychomotor stimulant properties, whereas pretreatment with morphine causes cross-sensitization of the locomotor effects of amphetamine. Here, we investigated whether pre-exposure to amphetamine also results in cross sensitization to morphine. Rats pretreated with amphetamine (5 x 2.5 mg/kg, i.p.) displayed neither short-term (3 days post-treatment) nor long-term (3 weeks post treatment) cross-sensitization of the locomotor effects of morphine (2 or 5 mg/kg, s.c.). Two other amphetamine pretreatment protocols (1 x 5 mg/kg, i.p. and 14 x 2.5 mg/kg, i.p.) also failed to induce cross-sensitization to morphine. In contrast, all amphetamine pretreatment regimens induced sensitization of the locomotor effects of amphetamine (1 mg/kg, i.p.) and pretreatment with morphine (14 x 10 mg/kg, s.c.) induced both short- and long-term sensitization of the locomotor effects of both morphine and amphetamine. These data suggest that the expression of sensitization of the locomotor effects of morphine and amphetamine, at least partially, involves distinct neuroadaptive phenomena. PMID- 10481840 TI - Behavioral and biochemical manifestations of mecamylamine-precipitated nicotine withdrawal in the rat: role of nicotinic receptors in the ventral tegmental area. AB - Brain mesolimbic dopamine (DA) neurons are considered critical for the dependence producing action of nicotine, and its stimulatory effect on behavior and DA neurotransmission appears largely mediated via nicotinic receptors (nAChRs) in the ventral tegmental area (VTA). The nAChR antagonist mecamylamine administered systemically in chronically nicotine-treated rats elicits a behavioral withdrawal syndrome concomitant with a reduced DA output in the nucleus accumbens (NAC). Here, we investigated the behavioral and biochemical consequences of intrategmental administration of mecamylamine in rats chronically infused with nicotine by means of minipumps for 14 days (9 mg/kg/day). Bilateral, intrategmental mecamylamine injections (1, 3 or 9 micrograms/0.5 microliter/side) dose-dependently increased abstinence signs such as gasps, teeth chatter, and reduced locomotor activity in nicotine-treated, but not in control animals. Moreover, a unilateral intrategmental injection of 9 micrograms mecamylamine reduced DA output in the ipsilateral NAC of chronically nicotine-treated rats, but not in control animals. Consequently, nAChRs in the VTA may be involved not only in the stimulatory effects of acute nicotine administration, but also in the withdrawal reaction following cessation of chronic nicotine treatment. PMID- 10481841 TI - Effects of flupenthixol and quadazocine on self-administration of speedball combinations of cocaine and heroin by rhesus monkeys. AB - The simultaneous i.v. administration of heroin and cocaine, called "speedball," is often reported clinically, and identification of effective pharmacotherapies for polydrug abuse is a continuing challenge. This study compared the effects of treatment using combinations of dopamine and opioid antagonists with each antagonist alone on speedball self-administration by rhesus monkeys. Speedballs (0.01 mg/kg/inj cocaine and 0.0032 mg/kg/inj heroin) and food (1 g banana pellets) were available in four daily sessions on a second-order schedule of reinforcement [FR4 (VR16:S)]. Monkeys were treated for 10 days with saline or ascending 1:10 dose combinations of the dopamine antagonist flupenthixol and the opioid antagonist quadazocine. The combination of flupenthixol (0.018 mg/kg/day) + quadazocine (0.18 mg/kg/day) significantly reduced speedball self administration in comparison to the saline treatment baseline (p < .05), whereas, the same doses of each antagonist alone had no significant effect on speedball maintained responding. Treatment with 0.018 mg/kg/day flupenthixol + 0.18 mg/kg/day quadazocine produced a 3-fold rightward shift in the speedball (3:1 cocaine-heroin combination) dose-effect curve. Food-maintained responding was similar during treatment with saline and with flupenthixol + quadazocine combinations. These findings suggest that medication mixtures designed to target both the stimulant and opioid component of the speedball combination, may be an effective approach to polydrug abuse treatment. PMID- 10481842 TI - PET imaging of dopamine D2 receptors with [18F]fluoroclebopride in monkeys: effects of isoflurane- and ketamine-induced anesthesia. AB - The purpose of the present study was to determine whether positron emission tomography (PET) studies in monkeys with the dopamine (DA) D2 receptor ligand [18F]fluoroclebopride (FCP) would be significantly influenced by anesthetic induction with isoflurane (approximately 5.0%) compared to induction with 10 mg/kg ketamine. Five experimentally-naive adult male cynomolgus monkeys (Macaca fascicularis) were trained to sit calmly in a primate restraint chair. Before the first PET scan, each monkey was anesthetized, by mask, with isoflurane. After complete sedation, the monkey was intubated and anesthesia was maintained throughout the PET study by isoflurane (approximately 1.5%). At least 1 month later, a second PET study was conducted in which anesthesia was induced with ketamine and maintained by isoflurane (approximately 1.5%). Irrespective of induction anesthetic, there was a high uptake of [18F]FCP and a linear rate of washout from the basal ganglia for all monkeys. There were also no differences in time to peak uptake (approximately 25 min), in clearance half-life (t1/2 = 140 164 min) or in D2 binding (distribution volume ratios of 2.48 vs. 2.50). These results indicate that induction anesthetic did not differentially affect D2 binding of [18F]FCP in monkeys. Furthermore, the low variability between studies indicates that [18F]FCP is an excellent ligand for longitudinal studies of D2 receptors in nonhuman primates. PMID- 10481843 TI - MDMA study. PMID- 10481844 TI - Is a single dose of MDMA harmless? PMID- 10481845 TI - [Cytokine imbalance in critically ill patients: SIRS and CARS]. AB - It remains difficult to treat severely ill patients, especially those who have sepsis and subsequent multiple organ dysfunction syndrome. We propose the hypothesis that the pathophysiology in the sequential sepsis and multiple organ dysfunction syndrome may be strongly related to the imbalance between inflammatory cytokines and antiinflammatory cytokines induced for the host defense to active neutrophils and endothelial cells. Thus we attempted to develop cytokine modulation therapy to normalize the cytokine balance in the host defense system. In this review, we elucidate the relationship between cytokine imbalance and SIRS/CARS in patients with severe burn injury. Furthermore, we examine the possible usage of G-CSF to amplify neutrophil function, and clarify the reasons why various innovative therapies against sepsis have failed. PMID- 10481846 TI - [Advances in metabolic and nutritional management of patients in critical care]. AB - Nutritional management often fails in critically ill patients such as trauma and sepsis because of their acutely progressive malnutrition and metabolic derangement being characteristic of such clinical settings. Nutritional strategies for these patients have been almost established by the methods of clinical epidemiology. A considerable amount of clinical evidences suggests that the following should be of benefit for the patients, enteral delivery of nutrition, a nutrition assessment including metabolic evaluation such as liver function or energy requirements, and supplementation of specialty nutrients. Our current data suggest that the metabolic evaluation should include measurement of energy expenditure, assessment of hepatic mitochondrial function, and measurement of the magnitude of body water deviations. Energy expenditure and hepatic mitochondrial function can be observed by the methods of indirect calorimetry and arterial ketone body measurement, respectively. These parameters are helpful to prevent overfeeding from the patients. We emphasize that application of a new method, body impedance spectrum analysis, is important and useful to evaluate the change of body water distributions resulting from metabolic derangement. PMID- 10481847 TI - [Recent advances in the management of severely burned patients]. AB - With recent advances in the systemic care of burns, patients with burns covering 80% of their body surface can frequently survive. The percentage of total body surface area burn for an expected 50% mortality rate has improved to 98% for children and 72% for adults in one burn center in the USA. From the results of 11 burn units in Tokyo, the mortality rate of burn patients with a prognostic burn index of 90-100 was 51.4%. The improvement is attributable to advances in the understanding of the pathophysiology of severe burns as a systemic inflammatory response syndrome. Improved cardiopulmonary management of extensive burns and respiratory tract burns has also played a part in reducing the mortality rate. Individualized fluid resuscitation programs based on hemodynamic monitoring have reduced the incidence of burn shock and acute renal failure. Early eschar excision and wound closure by immediate grafting have further reduced the mortality rate from extensive full-thickness burns. The use of bilayer artificial skin has improved the survival and cosmetic results of early eschar excision in patients with massive full-thickness burns. Cultured autologous epidermal sheets hold promise if used on an appropriate dermal bed. PMID- 10481848 TI - [Damage control surgery]. AB - Trauma patients who receive exsanguinating torso injuries often develop hypothermia, metabolic acidosis, and coagulopathy before death. A new strategy for trauma surgery has been developed to avoid the occurrence of these events and hence prevent trauma deaths. The strategy is called "damage control surgery" and consists of three maneuvers: a) damage control; b) restoration of physiologic stability; and c) definitive surgery. The goals of damage control are to: a) identify injuries; b) control ongoing hemorrhage; and c) control intestinal spillage. Damage control is followed by intensive care to restore the physiologic reserve. Once secondary resuscitation in the ICU is accomplished, planned reoperation should be performed to repair anatomic injuries. Planned reoperation is usually possible within 36 hours after the initiation of intensive care. Some patients who undergo damage control develop abdominal compartment syndrome characterized by increased intraabdominal pressure, increased peak airway pressure, decreased urine output, and decreased cardiac output. Early decompression surgery should be considered in such patients. PMID- 10481849 TI - [Progress in interventional radiology (IVR) in emergency medicine]. AB - IVR has attracted much attention in Japan over the past decade, and it is expected to be rapidly developed and widely used. Not surprisingly, IVR has already become essential in emergency medicine. This paper addresses IVR of the thoracic and abdominal areas and the pelvis in emergency medicine, in particular the recent IVR focus on the vasculature. CO2-DSA, which enables extravasation and easy detection of a arteriovenous or portal shunt, is also useful in the detection of bleeding in emergency situations. In trauma cases, TAE, originating from an expanded concept of "damage control," is commonly used to stop bleeding in order to perform surgical treatment or used during surgery. Occasionally the two applications are combined when appropriate. TAE is effective in controlling retroperitoneal bleeding resulting from pelvic fracture and parenchymatous organ injuries in the abdominal area. Treatment with stent-grafts, originally used to treat true aneurysms, has recently been used for treatment of injuries of the aorta or arteries and for entry closure due to aortic dissection. Furthermore, stent placement is expected to become an effective cure for organ ischemia, resulting from acute dissections. Is addition, in order to cure rupture of esophagus or gastric varices, such new treatments as TIPS and BRTO have increasingly been used, coupled with the conventional PTO treatment. Continuous regional arterial infusion of protease inhibitor, in an attempt to cure severe acute pancreatitis, significantly reduced the infectious rate at the necrotic lesion, and its resulting mortality rate. It is certain that great progress has been made in emergency medicine. We also should realize that it is desirable to be well versed even in new IVR. PMID- 10481850 TI - [The brain hypothermia therapy for prevention of vegetation after severe brain injury]. AB - We have presented a new concept of brain hypoxia oriented brain hypothermia treatments. All severe brain injury patients (148 cases) were GCS < 6. The masking brain hypoxia by brain thermo pooling, catecholamine surge induced cardiac dysfunction and intestinal vasodilatation, reduction of Hb-2.3 diphosphoglyserate were major target of initial treatment. These specific brain hypoxia was only controlled by brain hypothermia (34-32 degrees C), oxygen delivery > 800 ml/min. and AT-III > 100%. 2. The brain hypothermia were very successful to prevent masking brain hypoxia, selective radical attack to A10 dopamine nervous system, and brain edema. However, metabolic shift to lipid metabolism and lower growth hormone related immune crisis were recorded as a negative factors. Clinical results were so advanced. The mortality were 44 in 148 cases (30%), good recovery were 59 in 148 cases (40%), mild disability were 20 in 148 cases (13%) and vegetate state were only 15 in 148 cases (10%). The combination of brain hypothermia and replacement of cerebral dopamine were very successful to prevent the vegetation in severe brain injury. PMID- 10481851 TI - [Critical appraisal of treatments in emergency settings]. AB - A critical appraisal was carried out of treatment with high-dose epinephrine (HDE) in cardiopulmonary resuscitation (CPR), active compression and decompression CPR (ACD-CPR), military antishock trousers (MAST), and Continuous renal replacement therapy (CRRT) in the emergency setting, based on evidence based medicine. Although the pathophysiological rationale encourage the clinical application of these treatments to improve the long-term outcome (mortality), no clinical trial could confirm their benefit. Additionally, Japanese investigators have reported no clinical epidemiological study related to this issue. Thus a clinicoepidemiological study associated with treatments in critical settings is required in this country. PMID- 10481852 TI - [Telemedical support in emergency medicine]. AB - In July 1997 we began support of paramedics who would respond to emergency scenes using a telecommunications system based in the hospital emergency unit. However, since no telecommunications system that could be employed on the actual clinical site was available, we had to develop a new system. Our telecommunications system (TMS-6101, NIHON KOHDEN WELLNES Co., Tokyo, Japan) can transmit vital signs in real time and interactively transmit several types of moving images. Because the current infrastructure for radio communications cannot provide adequate telemedical support for paramedics, we are using the new system for clinical support between our unit and associated hospitals. Based on 57 cases of clinical telemedical intervention, the system has enabled high-quality decision making by specialists without the need for them to travel to the scene or transport X-rays films, ultrasonographic reports, or endoscopic results. If this newly developed telecommunications system is employed for telemedical interventions in medical facilities in remote areas or on remote islands, medical consultations for Japanese individuals overseas, night-time first aid in urban areas, and in disaster situations, the physicians on both side of the line will be able to obtain a wealth of timely information, greatly influencing outcome in both emergency and nonemergency cases. PMID- 10481853 TI - [Out-of-hospital cardiac arrest and the Utstein style: utilization of an international standardized format to evaluate surgical outcome]. AB - The Utstein style is the recommended guidelines for uniform reporting of data from out-of-hospital cardiac arrest. The Utstein style was, originally developed to analyze out-of-hospital cardiopulmonary arrest due to cardiac etiology. However trauma remains a major cause of out-of-hospital cardiac arrest and it is recommended that it also be reported accurately by the Utstein style. There exist some controversy regarding treatment methods such as military antishock trousers (MAST) or emergency room resuscitative thoracotomy for cardiac arrest due to trauma. MAST is has been shown to be beneficial experimentally in traumatic shock and cardiac arrest, but this has not been confirmed clinically and Mast is used rarely. Emergency room resuscitative thoracotomy is a common treatment for cardiac arrest due to trauma. There have been no survivors among those who experienced trauma-related cardiac arrest and showed no signs of life before transport to a hospital, although the survival rate of cardiac arrest due to penetrating chest injury is relatively high, clinical date should be accumulated following the Utstein style to determine the indications for these treatments. PMID- 10481854 TI - Spontaneous unruptured bilateral ectopic pregnancy: conservative tubal surgery. PMID- 10481855 TI - Surgical management of vaginal agenesis. AB - Vaginal agenesis is a rare anomaly that may result from a variety of different underlying diagnoses. The most important aspects of the surgical management of this condition are: correct diagnosis of both the underlying abnormality and its anatomy, documentation of any associated renal or skeletal anomalies, and proper psychological preparation of the young woman for any anticipated corrective surgery. Over the years, there have been many different techniques devised in an attempt to provide these women with a functioning vagina that approximates normal anatomy. The purpose of this article is to review the surgical options for the management of this anomaly. PMID- 10481856 TI - Use of misoprostol in first and second trimester abortion: a review. AB - The use of misoprostol in medical termination of first and second term pregnancies and cervical priming in surgically induced termination of pregnancies has been studied extensively. A survey is given on the available literature (MEDLINE to May 1998) on the usage as a single medication or in combination with mifepristone or methotrexate. A review is given on literature concerning side effects and complications. Misoprostol is a most promising, cheap, and effective agent, which does not need cool storage like other prostaglandins. The use of misoprostol as an abortifacient has, however, not been supported by the manufacturer. This leads to the situation (similar to mifepristone/RU 486) that it is used and researched, but probably will not be officially approved for this specific indication. PMID- 10481857 TI - Adnexal torsion and pulmonary embolism: case report and review of the literature. AB - The classical teaching was that twisted adnexa should be resected and not untwisted, so as not to increase the risk of pulmonary embolism (PE). A patient recently was seen who developed PE after adnexal resection. Because this complication followed the conventional management of salpingo-oophorectomy, the literature was examined for cases of adnexal torsion and PE to see if the operative management (untwisting vs. excision without untwisting) could be implicated as a contributing factor. Three hundred nine cases of adnexal torsion managed by untwisting and 672 cases treated by adnexectomy without detorsion (untwisting) were found. The incidence of PE after adnexal torsion was 0.2 percent, and this incidence was not increased when the adnexa were untwisted. Therefore, we conclude that detorsion of twisted adnexa does not increase the risk of PE, compared with excision without untwisting. PE does occur in cases in which adnexal resection is performed without untwisting. Thus, detorsion of twisted adnexa should be considered at laparoscopy or laparotomy without fear of increasing the incidence of PE. PMID- 10481858 TI - [Mental health and stress coping among hospital nurses by stress & stress-coping questionnaire]. AB - This study was aimed to evaluate the relationship between the mental health level and stress coping style in a group of female subjects in a public hospital. A self-administrated questionnaire including questions on nursing work, the General Health Questionnaire (GHQ) as well as the Stress and Stress Coping questionnaire (SSCQ) was used. The subjects of this study were 225 female nurses. The subjects' age and working experience (mean +/- SD) were 34.0 +/- 10.3 and 10.6 +/- 8.3 years, respectively. The results obtained were as follows: 1) Either of the following determinants was related to negative mental health: experience of 1-2 years; working in the operating room or in-patient department. 2) The subjects whose working experience was < 5 years or > or = 10 years had higher scores in the SSCQ. These subjects were among those who did not have higher scores in the GHQ. It was suggested that these subjects need to receive a guidance in stress coping. These findings suggest that because working conditions have a negative influence on mental health, educational system for both inexperienced and experienced nurses is needed to develop an effective stress-coping style in medical institutions. PMID- 10481859 TI - [Prevalence of dental erosion caused by sulfuric acid fumes in a smelter in Japan]. AB - The effect of occupational sulfuric acid exposure and other factors on teeth was explored in a cross-sectional study with blind dental examinations. Among 350 male workers in a copper-smelter in Japan, 28 had mild dental erosion with silky glazed opaque appearance of the enamel and/or shallow concavities on the enamel. While opaqueness was observed in 20 and concavities found in 11 workers, only 3 had both signs, suggesting that the concave lesions were often accompanied with enough remineralization to keep the dental surface gloss, possibly due to acid exposure at a low level. The cases had a history of working in an electrolytic refining plant (36%), significantly more than in the non-cases (14%, p < 0.05). Some significant differences were found between the cases and the non-cases: the cases were older, had less dental plaque, less gingivitis, and more frequent toothbrushing habits. No significant differences were observed in possibly related dietary habits such as several types of acidic drinks. It was concluded that the present cases with dental erosion were most probably associated with occupational acid exposure. The risk ratio of cross-sectional prevalence of dental erosion for those with a history of electrolytic refining plant work was 3.0 (95% CI: 1.3-6.7) compared with those without a history of acid exposure. Whether the present work environment can still develop new cases of dental erosion remains to be studied. PMID- 10481860 TI - [Acute massive hemorrhage and disseminated intravascular coagulation]. PMID- 10481861 TI - [Results of clinical trials in the treatment of acute myeloid leukemia in adults during a 7-year period]. AB - AIM: Comparison of effectiveness of induction regimens varying in intensity and maintenance variants in patients with acute myeloid leukemia (AML) included in a randomised multicenter trial. MATERIALS AND METHODS: Clinical trial 1 enrolled 185 AML patients. Vepesid-free induction was used in 85 patients (group 1), induction with vepesid--in 99 patients (group 2). 223 AML patients entered trial 2. Of them 37 patients were treated for 3 years 7 + 3 with daunorubicin in the dose 45 mg/m2 in the induction phase (group 1), 85 and 101 patients received 7 + 3 with daunorubicin for a year in the dose 45 mg/m2 and 60 mg/m2, respectively, (group 2 and 3). RESULTS: For group 1 in trial 1 the remission rate, early lethality, resistance were, respectively, 60, 20 and 20%, respectively. For group 2 in trial 1--66, 22 and 12%, respectively. 5-year recurrence-free survival reached 32 and 37% for group 1 and 2, respectively. For trial 2 relevant figures made up 75.5, 16.2, 8.1% for group 1; 60, 17.6 and 22.4% for group 2; 63, 20.8 and 16% for group 3, respectively. The 3.5-year recurrence-free survival in groups 1, 2 and 3, was 16, 46 and 50%, respectively. For both trials, the differences between the groups were insignificant. CONCLUSION: The results evidence that the treatment can be shorter (not 3 but 1 year or even 6 months), the induction more intensive (the dose of anthracycline antibiotics can be elevated up to 60 mg/m2 without a rise in early lethality). PMID- 10481863 TI - [Liposomal daunorubicin (daunosom) in the treatment of recurrent acute promyelocytic leukemia]. PMID- 10481862 TI - [Effectiveness of trans-retinoic acid in the treatment of acute promyelocytic leukemia: initial results of a multicenter study]. AB - AIM: Evaluation of trans-retinoic acid (ATRA) in combination with cytostatic drugs in treatment of acute promyelocytic leukemia (APL). MATERIALS AND METHODS: In a multicenter study APL was treated according to protocols APL 01.97 and APL 06.87 in 28 patients (14 males and 14 females, median age 36 years). RESULTS: Administration of ATRA in combination with standard program 7 + 3 (cytosine arabinoside 100 mg/m2 twice a day v.v. day 1-7, daunorubicin 60 mg/m2 v.v. day 1 3) induced a complete remission in 25 patients (90%). Early lethality was 10% (3 patients died). Resistant APL was not registered. Retinoid syndrome was diagnosed in 15 patients, one patient died. 2-year overall and recurrence-free survival made up 72 and 82%, respectively. CONCLUSION: ATRA combination with cytosine arabinoside and daunorubicin is a novel treatment of acute promyelocytic leukemia providing a high rate of complete remission and long-term survival. PMID- 10481864 TI - [Effectiveness of allogeneic bone marrow transplantation in patients with acute leukemia in complete remission and in patients with chronic myeloid leukemia during the chronic phase]. AB - AIM: To study clinical efficiency of allogenic bone marrow transplantation (ABMT) in patients with acute leukemia (AL) in the first remission and in patients with chronic myeloid leukemia (CML) in chronic phase; to analyse overall and recurrence-free survival in relation to the diagnosis and age. MATERIALS AND METHODS: 26 patients with AL and 24 ones with CML (29 males and 21 females) were subjected to ABMT within 10 last years. Median of age in AL and CML was 24.5 and 25.5 years, median of the time since the diagnosis was 9 and 13 months, respectively. Follow-up since the ABMT made up 67.5 months (31-107) and 38 months (6-108), respectively. Conditioning was made with cyclophosphamide (120 mg/kg) plus total radiation of the body (12 Gy) in 16 patients, myelosan (mileran) in a dose 16 mg/kg plus cyclophosphane (120 mg/kg) in 34 patients. The marrow was taken from HLA-identical sibs, enzygotic twins (5 recipients). Cytogenetic investigations were made in CML. The retention of the transplant was controlled by immunological and molecular tests. RESULTS: Among AL patients 50% are still alive. Probability of 80-month survival reached 55%, 110 months--42%. Probable recurrence-free survival was 78%. All the patients are in a complete clinico hematological remission. Among CML patients 75% are still alive. Of them 89% had a complete hematological remission, 72% are in a complete hematological and cytogenetic remission. Probable 110 month survival equals 75%, probability to survive without recurrence--52%. Early lethality (100 days) of toxic and infectious complications was as low as 10 and 6%, respectively. Frequency of lethal acute secondary disease was under 8%. CONCLUSION: ABMT made in AL patients during the first complete remission and in CML patients in the chronic phase brings about very good results which are much better than after routine cytostatic chemotherapy. PMID- 10481865 TI - [Clinical manifestations, diagnosis and course of Pneumocystis carinii pneumonia in patients with hematologic diseases]. AB - AIM: To characterize clinical, diagnostic and course features of pneumonia caused by Pneumocystis carinii (PC) in hematologic inpatients. MATERIALS AND METHODS: 27 patients with blood diseases were studied. 22 of them had acute respiratory insufficiency and 5 had unclear lung affection. The data from bronchoalveolar lavage (BAL), lung biopsy, serum tests for IgG, IgM anti-PC-antibodies were used for diagnosis of PC-pneumonia. RESULTS: PC-pneumonia was diagnosed in 8 of 27 patients. Clinical manifestations characteristic for PC-pneumonia were not found. In 5 patients the diagnosis was made on the evidence provided by BAL. Lymphocyte count in BAL was elevated to 27.7 +/- 8.7%. Open biopsy of the lung and transbronchial biopsy diagnosed PC-pneumonia in 2 and 1 patients, respectively. Previous BAL examinations failed to detect PC-pneumonia in 2 of them. In all the patients PC-pneumonia was associated with another infection (bacterial, cytomegaloviral). Histologically, the picture of the disease was determined by the severity of the lung affection or its complications. 5 of 8 patients failed treatment with trimethoprim-sulphamethoxazole and died. Marked respiratory insufficiency was registered at PC-pneumonia diagnosis in all the lethal cases. CONCLUSION: Clinical and x-ray pictures of PC-pneumonia in hemoblastosis patients are not specific. All such patients with symptoms of lung infection resistant to antibacterial and antifungal therapy should be examined for PC-pneumonia. PMID- 10481866 TI - [Multiple organ failure in patients with hemoblastosis during long-term follow-up of treatment]. AB - AIM: Investigation of the condition of the liver, gastrointestinal tract, heart in patients at the stage of clinicohematological remission after treatment for hemoblastosis (acute leukemia, Hodgkin's disease, non-Hodgkin's lymphoma). RESULTS: Marked functional-morphological changes were found: endomyocardial fibrosis, cardiopathy, pulmonary hypertension, chronic atrophic gastritis, colon lesions, dysbacteriosis, viral or drug-induced hepatitis. CONCLUSION: The above affections have developed because of the treatment: chemotherapy, radiotherapy, hemotransfusions, antibacterial drugs. They deteriorate life quality and require a special system of rehabilitation measures. PMID- 10481867 TI - [Interferon alfa-2b treatment of adult patients during early chronic phase of Ph1 positive chronic myeloid leukemia (initial report on a cooperative study, protocol CML-MIG-97)]. AB - AIM: Evaluation of clinical effectiveness of two regimens of induction therapy of an early chronic stage of Ph'-positive chronic myeloid leukemia including interferon-alpha 2b (intron-A, "Schering Plough") in a cooperative randomised trial on the protocol CML MIG-97. MATERIALS AND METHODS: 42 patients with chronic myeloid leukemia were treated either with intron-A in standard doses (5 IU/m2/day) alone or in combination with monthly 10-day courses of low-dose cytosine-arabinoside (10 mg/m2/twice a day). The effect was assessed by the international criteria of a complete and partial hematologic remission and the cytogenetic response. RESULTS: Intron-A therapy in standard doses produced a pronounced cytogenetic response in 28.6% of the patients. In low-dose interferon alpha-2b (reaferon and intron-A, 1-3 IU/m2/day) in combination with various regimens of chemotherapy only minimal cytogenetic response was achieved. CONCLUSION: A pronounced cytogenetic response in early chronic stage of CML to standard doses of intron A holds promise in prolongation of CML patients survival and design of new effective therapy programs. PMID- 10481868 TI - [Richter's syndrome: analysis of literature data and original observations]. AB - AIM: Review of literature data and original experience with Richter's syndrome. MATERIALS AND METHODS: 250 patients suffering from malignant lymphoproliferative diseases with blood and bone marrow lymphocytosis were observed. 8 (3.2%) of them developed diffuse large-cell lymphoma (criteria and classification of REAL). RESULTS: 5 of the above 8 patients demonstrated spontaneous regression of lymphocytosis. These cases may illustrate transformation (clonal progression) of one morphological variant of malignant non-Hodgkin's lymphoma into another one, more aggressive. For this rare variant of Richter's syndrome running with regression of lymphocytosis the term Richter-Lortolary syndrome is proposed. Lortolary was the first who revealed a decrease of lymphocytosis in Richter's syndrome. The studies of the genome structure, first of all, of immunoglobulin genes show that in Richter-Lortolary syndrome it is easier, to confirm monoclonality of the two tumors (lymphocytic and large-cell) than to reject it. However, the idea of transformation has not been confirmed morphologically yet. CONCLUSION: Development of diffuse large-cell lymphoma in the course of chronic lymphatic tumor does not always indicate terminal state, later stage of tumor progression and poor prognosis. PMID- 10481869 TI - [Acute and chronic renal failure in patients with myeloma]. AB - AIM: To analyze causes of reversible and irreversible renal failure in myeloma patients, lethal outcomes, treatment policy. MATERIALS AND METHODS: 43 myeloma patients with renal failure entered the trial. The replacement therapy consisted of hemodialysis, hemofiltration, hemodiafiltration. All the patients received full-dose polychemotherapy according to the programs M-2 and VAD. RESULTS: 69% of the patients retained normal renal function. 23% of the patients died. Partial recovery of renal function was observed in 1 patient who had to undergo dialysis once in 10-12 days. The patients survived from 5 days to 36 months (mean 20.6 months). The main causes of death in renal failure were sepsis (38%) and hemorrhagic stroke (14%). PMID- 10481870 TI - [Aplastic anemia and viral hepatitis (posthepatitic aplastic anemia)]. AB - AIM: To validate up-to-date policy of immunosuppressive therapy of patients with aplastic anemia (AA) which has developed either at acute or chronic phase of viral hepatitis. MATERIALS AND METHODS: 16 patients with AA detected within 6 months (9 patients) or 12-36 months (7 patients) after acute viral hepatitis received immunosuppressive therapy (antilymphocytic globulin, cyclosporin A, splenectomy). RESULTS: Posthepatitis aplastic anemias ran a severe and treatment resistant course in most cases. The immunosuppressive therapy produced a response in 44% of the patients. AA following acute viral hepatitis demands intensive and long-term immunosuppressive therapy with antilymphocytic globulin, cyclosporin A, splenectomy (in some cases) to achieve a persistent clinicohematological remission. PMID- 10481871 TI - [Experience in the use of allogeneic bone marrow transplantation in severe forms of aplastic anemia at the Byelorussian hematological center]. AB - AIM: Investigation of the response to antilymphocytic globulin (ALG) and transplantation of allogenic bone marrow (TABM) in patients with a severe form of aplastic anemia (AA). MATERIALS AND METHODS: 15 patients were treated for severe AA in 1995-1997. 8 patients of group 1 were given ALG/cyclophosphamide with subsequent TABM from HLA-identical sib donor. To prevent graft versus host reaction the patients took cyclosporin A (CSA) + prednisolone. Those who had no sib donor (7 patients) were treated with ALG/CSA (group 2). RESULTS: A complete remission at 7-30-month follow-up was observed in 6 patients of group 1. One patient died of infectious complications in rejection of the transplant and one patient died of acute graft versus host reaction. None of group 2 patients achieved the remission. One patient died of infectious complications. The others need continuous hemotransfusion therapy. CONCLUSION: In spite of high probability of early complications after TABM, TABM-subjected patients are more likely to achieve a complete remission and recurrence-free survival than those given immunosuppressive therapy alone. PMID- 10481872 TI - [Clinical significance and problems of treatment of patients with heparin-induced thrombocytopenia]. PMID- 10481873 TI - [Clinical manifestations and diagnosis of acute porphyria]. PMID- 10481874 TI - [Aleksandr Andreevich Rusanov (1909-1983)]. PMID- 10481875 TI - [Role of heat-shock proteins in cardioprotection of patients operated on for ischemic heart disease]. AB - Major stress protein HSP72 is known to participate in protecting cells and organisms against harmful factors including ischemia, trauma etc. Under study was the level of HSP72 in the myocardium of 32 patients with coronary disease operated in Military-medical academy. HSP72 was detected in probes of the right atria before and after pre-cardiopulmonary bypass in all cases induction of HSP72 was observed in 40% of patients, and directly correlated with the time of cardiopulmonary bypass and standing of the disease. The cardioprotective effect of the elevated pre-operational level of HSP72 was shown to be proportionate to the lower activity of cardiospecific enzymes, creatine phosphokinase (CK-MB). It is suggested that HSP72 is involved in the mechanism of cardioprotection during cardiopulmonary bypass. PMID- 10481876 TI - [Endoscopy in the diagnosis and treatment of gastroduodenal hemorrhage]. AB - An experience of work of the department of general surgery with the course of endoscopy is presented. The endoscopic diagnosis and treatment of 2931 patients for 6 years were performed for bleedings from the upper parts of the gastrointestinal tract. Errors in endoscopic diagnosis made 5%. The authors divided them into three groups: methodical (preparing the patients, methods of examination), diagnostic (assessment of the substrate and type of bleeding, probability of recurrences), tactical (medical policy, dynamics of observations). Hemostasis in endoscopic arrest may be complete or temporary (stable or unstable). The most effective methods are thought to be coagulation, clipping and a combination of the methods. Medicamentous means are less effective. The authors recommend to be aware of the endoscopic possibilities and not to try to stop bleeding by any means thereby wasting time. PMID- 10481877 TI - [Diagnosis of disorders of the motor function of the duodenum as a cause of chronic biliary pancreatitis]. AB - Great clinical material presented in the article concerns diseases of the pancreas. The authors prove that the leading role in the appearance of chronic pancreatitis is not infrequently played by disturbances of the motor function of the duodenum. The principal method of diagnosis is thought to remain the roentgenological examination. PMID- 10481878 TI - [How often is breast fibroadenomatosis asymptomatic?]. AB - The results of clinico-morphological examinations of 386 women having no complains of discomfort in the mammary glands are presented. Methods of palpation, mammography and morphological investigation have detected fibroadenomatosis without subjective symptoms of the disease in 38%, 67% and 78% of the patients correspondingly. This variant of the disease is called asymptomatic since the woman can not notice it and so it is out of the field of attention of oncologists. High incidence of asymptomatic fibroadenomatosis requires a revision of the current assessment of it as a disease because it does not take into account a variant of the disease when the patient has no complains of painful indurations in the mammary glands. PMID- 10481879 TI - [Transcutaneous determination of oxygen and carbon dioxide pressure in patients with obliterating atherosclerosis of blood vessels of the lower limbs]. AB - The method of transcutaneous determination of oxygen and carbon dioxide tension was used in examinations of healthy men (26 extremities) and 66 patients with obliterating atherosclerosis of the lower extremity arteries with different stages of the insufficient peripheral blood circulation. It was shown that oxygen tension in the foot skin was decreased in proportion to the decrease of the humero-malleolar index. In patients with obliterating atherosclerosis of the vessels and the II stage of insufficiency of blood circulation of the lower extremity the decrease of the humero-malleolar index and capillary blood flow of the foot skin did not result in a sharp disturbance of the gas regimen of the skin. In the III and IV stages of disturbances of blood circulation there occur failure of compensatory-adaptational mechanisms of tissue respiration accompanied not only by pronounced hypoxia but also by hypercapnia. PMID- 10481880 TI - [Bronchoscopic methods in complex diagnosis and treatment of burned patients with inhalation injuries]. AB - Chromobronchoscopy was first used in treatment of 37 burned patients with inhalation injuries for better visual manifestation of the injuries of the tracheobronchial tree mucosa. The trachea and bronchi mucosa was irrigated with a viral stain--0.25% aqueous solution of methylene blue. The intensity of staining the mucosa in light blue colour showed the true limits and depth of the injury. Resulting from the clinico-endoscopic examinations supplemented with findings of chromobronchoscopy, cytological and bacteriological analyses, a working classification of inhalation injuries in burned people was developed which allowed the adequate methods of local treatment during fiber bronchoscopy to be worked out taking into account the degree of thermochemical injuries of the respiratory pathways. It resulted in 19.7% lower lethality of burned patients. PMID- 10481881 TI - [Diagnosis and treatment of gastroesophageal reflux in children]. AB - The work is based on an analysis of results of treatment of 643 children with gastroesophageal reflux (operations were performed on 69 of them). Methods of anesthesia are described. Conservative treatment was used in correlation with the degree of reflux-esophagitis. Indications to operations were considered to be as follows: hiatal hernias, reflux-esophagitis in children with cerebral paralysis, failure of conservative treatment in the group of children having no anatomical causes of the reflux. A method of surgical treatment is proposed including a combination of fundoplication with the posterior gastropexy and fixation of the esophagus to the diaphragm at a distance of 3 cm from the gastric cuff (43 cases, no recurrences). In patients with a combination of gastroesophageal reflux with the esophagus stenosis resection of the altered portion was made followed by plasty with a colonic transplant or local tissues (10 patients with the diagnosed Barrett esophagus). PMID- 10481883 TI - [Surgical treatment of "difficult" gastric ulcers]. AB - In surgical treatment of "difficult" ulcers of the stomach including ulcers of the proximal part of the stomach, giant ulcers (more than 3 cm in diameter) and multiple ulcers it is expedient to use resection of the stomach with the formation of functionally active anastomosis. The choice of the operative method is determined by specific features of the ulcerous process, morphofunctional state of proximal parts of the gastrointestinal tract which determines the individual character of the operative intervention. The use of such operative interventions has reduced the postoperative lethality to 0.9%. Complex examinations have established excellent and good results (by the Visick scale) at remote terms after operation in 95.2% of the patients. PMID- 10481882 TI - [Prognosis of the course of suppurative-inflammatory surgical diseases of the abdominal cavity when using a fluorescent test for albumin]. AB - Fluorescent parameters named total and effective albumin concentrations (TA and EA, respectively) were used for prognosis of development of inflammatory surgical abdominal diseases: peritonitis, acute pancreatitis and some related states. Normal range of EA values is 35-55 g/L. At EA less than 8 g/L the mortality was 92%, and at EA more than 20 g/L the mortality was 5%. Very high mortality risk (up to 100% for patients above 50 years of age) was expected at narrow range of the parameter IT = TA/EA-1 values (from 0.7 to 0.8). Thus EA and IT could be taken as new parameters to estimate mortality and survival chances in acute inflammatory surgical diseases. PMID- 10481884 TI - [Method of closing the anterior and middle portions of the hard palate in uranoplasty]. AB - A new method of closing the anterior and middle portions in surgical treatment of cleft palate is proposed. The method allows obtaining the necessary reserve of plasty material, optimal adaptation of the wound surface margins and gives complete closing of the palate defects. The method proposed was used in operations on 6 patients with different forms of the cleft palate. Good results were obtained. PMID- 10481885 TI - [Prevention of prosthetic heart valve infection]. AB - Fourty patients with primary and secondary infective endocarditis of the native heart valves as well as patients with rheumatic heart diseases not complicated by infective endocarditis were operated upon in the clinic from 1996 till 1998. The prosthetic heart valves in which the tissue cuff was sutures by blanket suture along the perimeter with a silver thread were implanted to 25 patients. In the other 15 patients (a control group) prosthetics were made using unchanged construction of the prosthesis (without the silver thread). Patients with valve prostheses sutured with the silver thread had smooth postoperative period. They had no prosthetic endocarditis either at the early or at late postoperative periods. In 3 patients of the control group infective endocarditis of the valve prosthesis developed at different periods after operation. All of them were reoperated. The authors consider that defense of the tissue cuff of the prosthetic valve with the silver thread allows risk of the development of prosthetic endocarditis to be substantially reduced. In the cases described it was completely avoided. PMID- 10481886 TI - [Zenker's diverticulum]. AB - An experience with treatment of 14 women and 15 men aged from 41 to 83 years with Zenker's diverticulum was analyzed. Most liable to the disease were people at the age from 50 to 70 years. The average duration of the disease was 3-4 years. All the patients were subjected to improved surgical interventions: resection of the diverticulum (11 patients), operations under the control of the endoscopic technique (18 patients). Resections of the diverticulum were combined with cryopharyngeal myotomy. No complications were noted. There were no lethal outcomes. So, wider indications to operative treatment in patients with Zenker's diverticulum, endoscopic methods used at all stages of treatment as well as improved methods of performing the operation enabled the authors to obtain good immediate and long-term results. PMID- 10481887 TI - [Possibilities of diagnostic microlaparotomy in emergency surgery of organs of the abdominal cavity]. AB - A method of diagnosing acute diseases of abdominal organs in patients who had been subjected to operations on organs of the abdominal cavity is described. The method is efficient and can give better results of treatment of such patients. It allows the amount of exploratory laparotomies to be reduced. PMID- 10481888 TI - [Surgical treatment of patients with perforating gastroduodenal ulcers]. AB - Under analysis were the results of treatment of 267 patients with perforating gastroduodenal ulcers for the period from 1992 till 1997. Radical operations were fulfilled in 86% of the patients. Suture of the perforation and selective proximal vagotomy were performed in 118 patients with ulcers of the duodenum having no intraoperative signs of duodenostasis, in the others resection of the stomach was used. Palliative operations (suturing the perforation) were performed in 15% of the patients. General lethality was 3.7%, postoperative--3%. Excellent and good results were obtained in 95.5% of the patients. PMID- 10481890 TI - [Strategy of the surgical treatment of postoperative complications in patients with obstructive ileus of the colon of tumorous etiology]. AB - Under analysis were postoperative intraperitoneal complications and results of relaparotomies in 33 patients (13 women and 20 men) aged from 33 to 76 years with obturative ileus caused by stenosing cancer of different portions of the colon. High frequency of peritonitis in the general picture of postoperative complications was due to incompetent sutures of the colonic anastomoses applied during emergency and urgent operations in patients with sub- and decompensated forms of intestinal obstruction. The best variant of completion of radical operations in emergency surgery in this category of patients is the formation of a single- or double-trunk colostomy without a primary reestablishment of the continuity of the intestinal tube especially when stenosing cancer is localized in the left part of the colon. PMID- 10481889 TI - [Experience in the treatment of patients with gastroduodenal hemorrhage of different etiology: based on materials of an air ambulance service]. AB - The article is devoted to an analysis of main causes of the development of acute gastroduodenal bleedings, methods of surgical treatment in small town and district hospitals with participation of consulting surgeons of aerial ambulance. Postoperative outcomes are also considered. The authors made a conclusion that active surgical management and performing primary radical operations on the spot allowed lethality to be reduced to 6.6%. However, the optimum policy is thought to be transportation of the patients to specialized regional medical institutions after temporary adequate endoscopic arrest of bleeding and further treatment of them at sufficiently equipped centers with highly trained specialists. PMID- 10481891 TI - [Surgical treatment of true muscle hernias of the extremities]. AB - An original method of plastic closing the fascial defects with the polymer tissue was used in 5 patients with true muscular hernias of the extremities. In all cases good anatomo-functional and cosmetic results were obtained. PMID- 10481893 TI - [Removal of a foreign body from the right heart ventricle without diversion of blood circulation]. PMID- 10481892 TI - [Continuous inhalation administration of surfactant-BL for control of adult respiratory distress syndrome after reconstruction of the abdominal aorta]. AB - A 62-year-old patient with Leriche's syndrome and critical ischemia of the low extremities has undergone the surgery of aortofemoral grafting. A patient has developed the severe ARDS on the second day of reperfusion (bilateral diffuse infiltrates, PO2/FiO2 < 100, lung injury score was 3). Different ways of administration of bovine surfactant (SURFACTANT BL, Russia) were used during the treatment. Total application time was 84 hours, total dose was 4000 mg (50 mg/kg). Considerable improvement of lung function occurred after start of the continuous inhalation of surfactant with a constant rate of 63 mg/h. Two hours after this step PO2/FiO2 reached 400 mm Hg and remained stable 12 hours more, up to the end of surfactant administration. The patient was soon successfully extubated and discharged on the 25th day after surgery. It is supposed that in spite of a small total dose of bovine surfactant the success was achieved due to an early start of the surfactant replacement, continuous inhalation and its definite rate. PMID- 10481894 TI - [A rare localization of the entrance area in stab/cut wounds of the abdomen with liver injury]. PMID- 10481895 TI - [One-stage surgery in calculous cholecystitis and renal cancer]. PMID- 10481896 TI - [Endoscopic treatment of a woman patient with subdiaphragmatic abscess]. PMID- 10481897 TI - [2 unusual cases of bezoars of the intestine]. PMID- 10481898 TI - [Acute intestinal obstruction caused by mesenchymoma]. PMID- 10481899 TI - [2 case reports of inflammatory tumors of the ileocecal part of the intestine]. PMID- 10481900 TI - [A rare complication in hernia of the anterior abdominal wall]. PMID- 10481901 TI - [Some remarks concerning the article by I.P. Dudanov and coauthors, "Debatable questions regarding acute simple appendicitis and ways of reducing incidence of unnecessary appendectomies"]. PMID- 10481902 TI - [Sepsis, terminology and nature (impressions and thoughts of a participant of a consensus conference)]. PMID- 10481903 TI - [Fatal wound of A.S. Pushkin. Going back to autographs]. PMID- 10481904 TI - [Surgeons -- "architects" of the Academy]. PMID- 10481905 TI - Narcolepsy: a key role for hypocretins (orexins) PMID- 10481906 TI - Novel DNA polymerases offer clues to the molecular basis of mutagenesis. PMID- 10481907 TI - Reorganizing the organizer 75 years on. PMID- 10481908 TI - Mice with reduced NMDA receptor expression display behaviors related to schizophrenia. AB - N-methyl-D-aspartate receptors (NMDARs) represent a subclass of glutamate receptors that play a critical role in neuronal development and physiology. We report here the generation of mice expressing only 5% of normal levels of the essential NMDAR1 (NR1) subunit. Unlike NR1 null mice, these mice survive to adulthood and display behavioral abnormalities, including increased motor activity and stereotypy and deficits in social and sexual interactions. These behavioral alterations are similar to those observed in pharmacologically induced animal models of schizophrenia and can be ameliorated by treatment with haloperidol or clozapine, antipsychotic drugs that antagonize dopaminergic and serotonergic receptors. These findings support a model in which reduced NMDA receptor activity results in schizophrenic-like behavior and reveals how pharmacological manipulation of monoaminergic pathways can affect this phenotype. PMID- 10481909 TI - Narcolepsy in orexin knockout mice: molecular genetics of sleep regulation. AB - Neurons containing the neuropeptide orexin (hypocretin) are located exclusively in the lateral hypothalamus and send axons to numerous regions throughout the central nervous system, including the major nuclei implicated in sleep regulation. Here, we report that, by behavioral and electroencephalographic criteria, orexin knockout mice exhibit a phenotype strikingly similar to human narcolepsy patients, as well as canarc-1 mutant dogs, the only known monogenic model of narcolepsy. Moreover, modafinil, an anti-narcoleptic drug with ill defined mechanisms of action, activates orexin-containing neurons. We propose that orexin regulates sleep/wakefulness states, and that orexin knockout mice are a model of human narcolepsy, a disorder characterized primarily by rapid eye movement (REM) sleep dysregulation. PMID- 10481910 TI - The Drosophila caspase inhibitor DIAP1 is essential for cell survival and is negatively regulated by HID. AB - Drosophila Reaper (RPR), Head Involution Defective (HID), and GRIM induce caspase dependent cell death and physically interact with the cell death inhibitor DIAP1. Here we show that HID blocks DIAP1's ability to inhibit caspase activity and provide evidence suggesting that RPR and GRIM can act similarly. Based on these results, we propose that RPR, HID, and GRIM promote apoptosis by disrupting productive IAP-caspase interactions and that DIAP1 is required to block apoptosis inducing caspase activity. Supporting this hypothesis, we show that elimination of DIAP1 function results in global early embryonic cell death and a large increase in DIAP1-inhibitable caspase activity and that DIAP1 is still required for cell survival when expression of rpr, hid, and grim is eliminated. PMID- 10481911 TI - Disruption of the sarcoglycan-sarcospan complex in vascular smooth muscle: a novel mechanism for cardiomyopathy and muscular dystrophy. AB - To investigate mechanisms in the pathogenesis of cardiomyopathy associated with mutations of the dystrophin-glycoprotein complex, we analyzed genetically engineered mice deficient for either alpha-sarcoglycan (Sgca) or delta sarcoglycan (Sgcd). We found that only Sgcd null mice developed cardiomyopathy with focal areas of necrosis as the histological hallmark in cardiac and skeletal muscle. Absence of the sarcoglycan-sarcospan (SG-SSPN) complex in skeletal and cardiac membranes was observed in both animal models. Loss of vascular smooth muscle SG-SSPN complex was only detected in Sgcd null mice and associated with irregularities of the coronary vasculature. Administration of a vascular smooth muscle relaxant prevented onset of myocardial necrosis. Our data indicate that disruption of the SG-SSPN complex in vascular smooth muscle perturbs vascular function, which initiates cardiomyopathy and exacerbates muscular dystrophy. PMID- 10481912 TI - Activation of store-operated Ca2+ current in Xenopus oocytes requires SNAP-25 but not a diffusible messenger. AB - Depletion of Ca2+ stores in Xenopus oocytes activated entry of Ca2+ across the plasma membrane, which was measured as a current I(soc) in subsequently formed cell-attached patches. I(soc) survived excision into inside-out configuration. If cell-attached patches were formed before store depletion, I(soc) was activated outside but not inside the patches. I(soc) was potentiated by microinjection of Clostridium C3 transferase, which inhibits Rho GTPase, whereas I(soc) was inhibited by expression of wild-type or constitutively active Rho. Activation of I(soc) was also inhibited by botulinum neurotoxin A and dominant-negative mutants of SNAP-25 but was unaffected by brefeldin A. These results suggest that oocyte I(soc) is dependent not on aqueous diffusible messengers but on SNAP-25, probably via exocytosis of membrane channels or regulatory molecules. PMID- 10481913 TI - Store-operated Ca2+ entry: evidence for a secretion-like coupling model. AB - The elusive coupling between endoplasmic reticulum (ER) Ca2+ stores and plasma membrane (PM) "store-operated" Ca2+ entry channels was probed through a novel combination of cytoskeletal modifications. Whereas coupling was unaffected by disassembly of the actin cytoskeleton, in situ redistribution of F-actin into a tight cortical layer subjacent to the PM displaced cortical ER and prevented coupling between ER and PM Ca2+ entry channels, while not affecting inositol 1,4,5-trisphosphate-mediated store release. Importantly, disassembly of the induced cortical actin layer allowed ER to regain access to the PM and reestablish coupling of Ca2+ entry channels to Ca2+ store depletion. Coupling is concluded to be mediated by a physical "secretion-like" mechanism involving close but reversible interactions between the ER and the PM. PMID- 10481914 TI - Nuclear shuttling of yeast scaffold Ste5 is required for its recruitment to the plasma membrane and activation of the mating MAPK cascade. AB - Localization of Ste5 to GP at the plasma membrane is essential for transmission of the pheromone signal to associated MAP kinase cascade enzymes. Here, we show that this crucial localization requires prior shuttling of Ste5 through the nucleus. Ste5 shuttles through the nucleus constitutively during vegetative growth. Pheromone enhances nuclear export of Ste5, and this pool translocates vectorially to the cell periphery. Remarkably, Ste5 that cannot transit the nucleus is unable to localize at the periphery and activate the pathway, while Ste5 with enhanced transit through the nucleus has enhanced ability to localize to the periphery and activate the pathway. This novel regulatory scheme may ensure that cytoplasmic Ste5 does not activate downstream kinases in the absence of pheromone and could be applicable to other membrane-recruited signaling proteins. PMID- 10481915 TI - Epigenetic spreading of the Drosophila dosage compensation complex from roX RNA genes into flanking chromatin. AB - The multisubunit MSL dosage compensation complex binds to hundreds of sites along the Drosophila single male X chromosome, mediating its hypertranscription. The male X chromosome is also coated with noncoding roX RNAs. When either msl3, mle, or mof is mutant, a partial MSL complex is bound at only approximately 35 unusual sites distributed along the X. We show that two of these sites are the roX1 and roX2 genes and postulate that one of their functions is to provide entry sites for the MSL complex to recognize the X chromosome. The roX1 gene provides a nucleation site for extensive spreading of the MSL complex into flanking chromatin even when moved to an autosome. The spreading can occur in cis or in trans between paired homologs. We present a model for how the dosage compensation complex recognizes X chromatin. PMID- 10481916 TI - Structures of two repeats of spectrin suggest models of flexibility. AB - Spectrin is a vital component of the cytoskeleton, conferring flexibility on cells and providing a scaffold for a variety of proteins. It is composed of tandem, antiparallel coiled-coil repeats. We report four related crystal structures at 1.45 A, 2.0 A, 3.1 A, and 4.0 A resolution of two connected repeats of chicken brain alpha-spectrin. In all of the structures, the linker region between adjacent units is alpha-helical without breaks, kinks, or obvious boundaries. Two features observed in the structures are (1) conformational rearrangement in one repeat, resulting in movement of the position of a loop, and (2) varying degrees of bending at the linker region. These features form the basis of two different models of flexibility: a conformational rearrangement and a bending model. These models provide novel atomic details of spectrin flexibility. PMID- 10481918 TI - Antioxidant status and lipid peroxidation in human feto-placental unit. AB - The human feto-placental unit being in the environment of the materno-fetal circulation is very much susceptible to the danger of reactive oxygen species induced oxidative damage. The present study reports the status of various antioxidants in the feto-placental system at birth. The activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase as well as concentrations of non-enzymic antioxidants viz, ascorbic acid, alpha-tocopherol, bilirubin and glutathione have been determined in term placental brush border membrane (BBM) and umbilical cord (UC) blood. The extent of lipid peroxidation in developing and term BBM and UC blood has been studied by estimation of malondialdehyde and conjugated dienes, both being the end products of lipid peroxidation. The results indicate the presence of a protective mechanism against oxygen toxicity in the feto-placental system at the time of parturition. PMID- 10481917 TI - Structure of the alpha-actinin rod: molecular basis for cross-linking of actin filaments. AB - We have determined the crystal structure of the two central repeats in the alpha actinin rod at 2.5 A resolution. The repeats are connected by a helical linker and form a symmetric, antiparallel dimer in which the repeats are aligned rather than staggered. Using this structure, which reveals the structural principle that governs the architecture of alpha-actinin, we have devised a plausible model of the entire alpha-actinin rod. The electrostatic properties explain how the two alpha-actinin subunits assemble in an antiparallel fashion, placing the actin binding sites at both ends of the rod. This molecular architecture results in a protein that is able to form cross-links between actin filaments. PMID- 10481919 TI - Effect of flavonoids on protease activities in human skeletal muscle tissue in vitro. AB - The effect of a representative range of flavonoid compounds on the activities of a range of proteolytic enzyme types in human skeletal muscle tissue was determined in vitro. All of the proteases showed some degree of inhibition by all of the flavonoid compounds investigated, with IC50 values (concentration of flavonoid required to inhibit initial activity of muscle proteases by 50%) for various enzyme/flavonoid combinations ranging from approx. 50 micromol/l to 2 mmol/l. These data demonstrate that in addition to their well described antioxidant characteristics, flavonoids also possess novel antiproteolytic inhibitory activity in vitro, which may in part explain some of the beneficial effects of these plant derived compounds in vivo. PMID- 10481920 TI - A pilot study of garlic consumption shows no significant effect on markers of oxidation or sub-fraction composition of low-density lipoprotein including lipoprotein(a) after allowance for non-compliance and the placebo effect. AB - Our aim was to define any effects of confirmed garlic supplementation on the resistance of low density lipoprotein (LDL) to oxidation, on LDL sub-fraction composition including levels of lipoprotein(a), and on levels of circulating antibody to oxidised LDL, variables of interest in relation to cardiovascular risk. Additional tests were performed on samples collected from a double blind, randomised 6-month parallel trial in which 900 mg Kwai garlic or placebo was taken by moderately hypercholesterolaemic volunteers. Final data was analysed for 20 garlic and 11 placebo subjects with compliance of at least 75% as determined by repeat tablet counting. EDTA plasma stabilised by sucrose was stored at -70 degrees C for up to 12 months. Lipids and apolipoproteins were determined by standard methods, lipoprotein(a) by an ELISA method and LDL fraction composition by non-gradient gel electrophoresis. Oxidative resistance of LDL purified after isolation by density gradient centrifugation was assessed from oxidative resistance to copper ions determined spectrophotometrically, antibodies to oxidised LDL were determined by a microtitre plate assay and vitamin E content of plasma by HPLC. Overall lipid/lipoprotein profiles including lipoprotein(a) were unchanged as with the parent group. LDL composition showed a trend to less dense material in both placebo and garlic groups, all differences not significant. Lag time as a marker of oxidative resistance also increased in both groups, without change in vitamin E content, all differences not significant and consistent with a placebo effect. Levels of antibodies to oxidised LDL were unchanged. The results of this study do not support the hypothesis that dietary garlic supplementation decreases the susceptibility of isolated LDL to oxidation and that patterns of LDL tractions in plasma might be involved. Levels of lipoprotein(a) in plasma were also not changed. Other mechanisms of cardiovascular benefit are however not excluded. PMID- 10481921 TI - Link between glycation and lipoxidation in red blood cells in diabetes. AB - Oxidative stress is postulated to be increased in patients with diabetes mellitus. Glycation enhanced by elevated glucose concentrations may induce the formation of oxygen-derived free radicals (OFRs). OFRs would cause oxidative damage to endogenous molecules, including cholesterol. Accumulating evidence suggests that oxidative cell injury caused by OFRs contributes to the development of both macroangiopathy and microangiopathy in diabetes. Our previous studies have shown that 7-keto cholestadien is one of the major products of cholesterol peroxidation in diabetic erythrocyte membrane and its levels correlate with hemoglobin Alc (HbAlc) values. We have newly identified 3-cholesten-6-one, one of the minor products of cholesterol peroxidation, in it. The aim of our study is to investigate whether 3-cholesten-6-one levels also correlate with HbAlc values. Levels of 3-cholesten-6-one were assessed in erythrocyte membrane lipid by monitoring peak areas of 3-cholesten-6-one to cholesterol with gas chromatography mass spectrometry. The peak area ratio of 3-cholesten-6-one to cholesterol was used as a marker of cholesterol peroxidation. The HbAlc value, an index of both glycemic stress and glycation, was measured by high-performance liquid chromatography. In this study, we evaluated 33 diabetic and 29 healthy subjects, matched for age (59.3+/-14.5 vs. 57.3+/-13.7 years, mean+/-S.D.) and sex (15 males and 14 females vs. 16 males and 17 females). There were both significantly raised HbAlc levels (4.6+/-0.8 vs. 8.3+/-2.4%, P<0.001) and significantly increased ratios of 3-cholesten-6-one to cholesterol (0.2+/-0.4 vs. 21+/-1.8, P<0.001) in diabetic patients compared to control subjects. A good correlation between HbAlc levels and ratios of 3-cholesten-6-one to cholesterol was found in participants (r = 0.75, P<0.001, y = 0.46x-1.8). This suggests that an oxidative stress exists in diabetes and the link between glycation and lipoxidation is found in diabetic red blood cell. PMID- 10481922 TI - Thermodynamic characterisation of the mutated isoenzyme A of beta-N acetylhexosaminidase in GM2-gangliosidosis B1 variant. AB - Here we report the determination of the activation energies of the plasma isoenzymes of beta-N-acetylhexosaminidase (Hex, EC 3.2.1.52), isolated by chromatography in DEAE-cellulose, using the neutral chromogenic substrate 3,3'dichlorophenylsulfonphthaleinyl-N-acetyl-beta-D-glucosaminide. The activation energy of mutated Hex A isoenzyme (Ea approximately 71.5 kJ/mol) from a patient with GM2-gangliosidosis B1 variant, homozygote for the G533-->A (Arg178His) mutation, was significantly higher than that of normal Hex A (Ea approximately 41.8 kJ/mol) and analogous to that of Hex B isoenzyme (Ea approximately 75.1 kJ/mol). The determination of this thermodynamic variable of Hex in different biological specimens could allow for a straightforward biochemical characterisation of the GM2-gangliosidosis B1 variant. PMID- 10481923 TI - Determination of copper, zinc, and selenium in human plasma and urine samples by potentiometric stripping analysis and constant current stripping analysis. AB - Potentiometric stripping analysis and constant current stripping analysis are proposed as routine methods for analysis of copper, zinc and selenium in plasma and urine samples. The analytical performance of these methods is comparable with that reported for atomic absorption spectrometry. However the low cost, greater simplicity of the apparatus, and the facility of execution make this methodology a valid candidate for routine application in Clinical Chemistry laboratories. PMID- 10481924 TI - Sialic acid content of LDL and lipoprotein metabolism in combined hyperlipidemia and primary moderate hypercholesterolemia. AB - Low density lipoprotein (LDL) with low sialic acid content has been shown to cause intracellular lipid accumulation and therefore is suggested to be atherogenic. We investigated the sialic acid content of total LDL and its subfractions, and their relations to lipoprotein kinetics in 22 subjects with primary moderate hypercholesterolemia (IIa) and in 21 subjects with combined hyperlipidemia (IIb) matched for age, sex, BMI and the frequency of coronary artery disease. Sialic acid to protein ratio decreased gradually from VLDL and IDL to light and dense LDL and HDL, but was high in very dense LDL probably due to presence of Lp(a). Sialic acid to apo B ratio was significantly lower in dense and very dense subfractions of IIb than IIa. The sialic acid/apo B ratios of dense and very dense LDL subfractions were interrelated and were negatively associated with their cholesterol and triglyceride concentrations, and with the transport rate (TR) for dense LDL apo B. The only metabolic variable differing between the groups was the TR for dense LDL apo B, which was significantly higher in IIb vs. IIa. In addition, the TR for dense LDL apo B was positively associated with the esterification percentage of LDL cholesterol. In conclusion, low sialic acid content in dense and very dense LDL subfractions was associated with enhanced TR for LDL apo B and type IIb dyslipidemia. PMID- 10481925 TI - Lymphocyte glutathione levels in acute leukemia. AB - The current interest in the clinical significance of the glutathione estimation in leukemia is due to its widespread effect on the cell constituent and cell function of the hematopoietic system. It helps and protects the cells against free radicals and reactive oxygen products. The present study was conducted to estimate glutathione levels in lymphocytes of 20 patients with acute leukemia before and after chemotherapy to observe the relation of glutathione level to response to chemotherapy. Twenty age and sex matched healthy volunteers served as control. In acute myeloid leukemia, the levels were almost double than that of controls (P<0.001). In acute lymphoid leukemia they were 2.5 times of the control. Lymphocyte glutathione levels was higher in active phase of disease than in remission. Lymphocyte glutathione level could act as a marker of leukemic activity and may help to predict onset of relapse. But, it is not only the determinant of response to chemotherapy. PMID- 10481926 TI - Relationship between the apolipoprotein E and angiotensin-converting enzyme genotypes and LDL particle size in Japanese subjects. AB - We investigated whether the apolipoprotein E (apoE) and the angiotensin converting enzyme (ACE) genotypes contribute to the variance in low-density lipoprotein (LDL) particle size in Japanese subjects (n = 136; M/F= 106/30). ACE polymorphism was associated with neither LDL size nor individual lipid levels. In contrast, the subjects with the epsilon2 allele of the apoE genotype had significantly lower levels of total cholesterol (P = 0.002) and LDL cholesterol (P = 0.004) compared with those without the epsilon2 allele. The subjects with the epsilon4 allele had a significantly smaller LDL particle size than those without the epsilon4 allele (P = 0.012). Separate analyses of the male subjects showed similar associations. A stepwise regression analysis revealed the epsilon4 allele to be an independent contributing variable that could affect LDL particle size. Our results suggest that the apoE genotype is associated with the development of atherosclerotic disease, since the epsilon2 and epsilon4 alleles relate to a decrease in LDL cholesterol levels and a decrease in LDL particle size, respectively. PMID- 10481927 TI - C-erbB-2 expression and its relationship with ploidy, p53 abnormalities and epidermal growth factor receptor content in human non-small cell lung cancer. AB - This study attempts to clarify the role of c-erbB-2 overexpression in human non small cell lung cancer (NSCLC) and relate it with the p53 alterations, DNA index (D.I.) and epidermal growth factor receptor (EGFR) content in sixty four patients with NSCLC. c-erbB-2 and EGFR quantification were carried out from tissue homogenates using quantitative ELISA procedures. p53 alterations were determined by immunohistochemical (IHC) detection with the monoclonal antibody DO-7 and analysis for p53 mutations on exons 4 to 8 by single strand conformation polymorphism (SSCP). The D.I. was performed by flow cytometry. c-erbB-2 hyperexpression was found in 13 of 58 LC (22%), and it was closely associated with hyperdiploid tumors (D.I. >1.3; P = 0.00). The p53 abnormalities detected by SSCP were statistically more frequent in hyperdiploid tumors (16/25; P = 0.015) than in diploid ones (8/30). No relationship between the results of IHC p53 and SSCP was found. The patients with c-erbB-2 hyperexpressing tumors were prone to have frequent relapses (P = 0.03), although the patients with hyperdiploid NSCLC are the ones with the highest relapse rate (P = 0.008). From the results obtained in this study the following conclusions can be drawn: (a) c-erbB-2 hyperexpressing NSCLC are associated with abnormalities in other biological markers and with a greater rate of relapses; (b) SSCP seemed to be more specific that IHC to detect p53 molecular abnormalities; and (c) the D.I. is the parameter more tightly related with relapse. PMID- 10481928 TI - A variation in Bone Alkaline Phosphatase levels that correlates positively with bone loss and normal levels of aminoterminal propeptide of collagen I in girls with anorexia nervosa. AB - Anorexia nervosa (AN) is a very extended pathology among adolescent girls nowadays. These patients show a high degree of osteopenia; hence, study of their bone remodelling is of great interest. Serum bone alkaline phosphatase (bAP) and aminoterminal propeptide of procollagen I (PINP) provide good sensitivity in the analysis of bone alterations in postmenopausal osteoporosis. The aim of this study was to compare the usefulness of bAP and PINP in the study of bone remodelling in AN, and their possible correlation with the degree of osteopenia in this pathology. In order to help in the interpretation of the results, levels of the beta-isomer of urinary carboxyterminal propeptide of collagen I (beta-CTX) have also been included. Serum bAP (IRMA) Tandem R-Ostase, Hybritech), PINP (RIA, Orion Diagnostica) and CTX (CrossLaps ELISA, Osteometer) were determined in 41 girls with AN, aged 18.5+/-2.2 years (mean+/-SD) and in 31 healthy control women, aged 19+/-2.3 years. Bone mineral density (BMD) in lumbar spine was measured by DEXA in the AN group. We found that 41 of the 43 patients had BMD z-scores under 2. No significant differences were found in the levels of serum bAP nor in PINP and beta-CTX levels between controls and patients, although values in the AN group were highly variable. All the BMD z-score values were negative, and their absolute value correlates positively with bAP (P = 0.0279) and almost with beta CTX (P = 0.0921) but not with PINP (P = 0.4627). Bone AP correlates with PINP in control girls (P = 0.017), but not in the AN group (P = 0.3573). Patients with AN were divided into three groups according to their levels of bAP: low (I), normal (II) or high (III). Patients with the highest bAP levels also presented the highest increase in bone resorption, according to their beta-CTX levels, and the highest degree of osteopenia. However, values of PINP were similar in the three groups of patients. The bAP/beta-CTX ratios in subgroups I, II and III of AN patients were 0.035, 0.065 and 0.073, a finding that suggests that bAP is not indicating the real degree of bone mineralization in these patients, because it is a contradiction that the formation/resorption ratio should be higher in the patients who have the highest bone loss. These results could suggest that bone loss in AN is produced by an increase in bone resorption (beta-CTX), without variations in bone matrix formation (PINP); bAP levels are a good marker in the follow-up of osteopenia degree, but not a real indicator of bone mineralization, a similar situation to that of osteomalacia. PMID- 10481929 TI - The detection of a unique antigen associated with papillary thyroid carcinoma. AB - We produced antibodies against a thyroid papillary carcinoma homogenate (PCAb) and analyzed antigens recognized by this antibody using western blotting. Fifty four thyroid tissue specimens and 6 control tissue specimens obtained from non thyroid carcinoma (gastric tissue, colon and liver) were analyzed. Consequently, an antigen of 40 kDa in size was found in 16 of 16 (100%) of papillary thyroid carcinoma from primary lesions and in 2 of 2 (100%) papillary thyroid carcinoma from metastatic foci, whereas it was not detected in thyroid tissue samples from follicular carcinoma, anaplastic carcinoma, follicular adenoma, adenomatous goiter, Graves' disease and normal thyroid tissues. The reactivity of thyroglobulin antiserum (TgAb) to this 40 kDa antigen was tested by western blotting and showed that TgAb did not appear to recognize the 40 kDa antigen. Moreover PCAb, after treatment with Tg, still reacted with this 40 kDa antigen. Therefore, this 40 kDa antigen might be different from Tg. Furthermore, to inspect the structure of this antigen, the effect of some chemicals and enzymes such as 2-mercaptoethanol, sodium dodecyl sulfate, ethanol and protease on the reactivity of PCAb to the 40 kDa antigen were analyzed. The results of these experiments suggested that this 40 kDa antigen may have a peptide structure. To our knowledge, the finding reported here represents the first demonstration of the protein specifically present in papillary thyroid carcinoma. Further investigations should elucidate the characteristics of this antigen and may contribute to definitive diagnosis of thyroid carcinoma as well as improving the understanding of the mechanisms involved in developing the thyroid carcinoma. PMID- 10481930 TI - Identification of compound heterozygous mutations (G188E/W382X) of lipoprotein lipase gene in a Japanese infant with hyperchylomicronemia: the G188E mutation was newly identified in Japanese. AB - We herein report a case of a 5-month-old Japanese female (patient AN) with fasting hyperchylomicronemia due to a primary lipoprotein lipase (LPL) deficiency. Patient AN was compound heterozygous for a missense mutation (GG818G- >GAG/Gly188-->Glu; G188E) in exon 5 and a nonsense mutation (TGG1401-->TGA/Trp382 ->Stop; W382X) in exon 8 of the LPL gene. This resulted in less than 10% of the control levels for both the LPL activity and immunoreactive LPL mass in the postheparin plasma. A G188E mutation was thus identified for the first time in a Japanese, and the haplotype of this G188E allele was different from that of the G188E alleles identified in other ethnic groups. This additional mutation might be useful for early diagnosis of LPL gene aberrations in Japanese patients with fasting hyperchylomicronemia. PMID- 10481931 TI - Cloning and sequence of partial cDNAs encoding the human type V and VI adenylyl cyclases and subsequent RNA-quantification in various tissues. AB - Adenylyl cyclases (Acs) and VI are the predominant form of Acs in mammalian heart where they are part of the beta-adrenergic pathway. Up to now, the sequences for both enzymes from human tissues have not yet been reported. We investigated the mRNA expression AC V and VI in human colon, heart, liver, lung and MNL. Partial cDNAs of human types V and VI Acs were amplified by PCR using oligonucleotides derived from the cytoplasmatic domain sequences of the corresponding enzymes from dog heart. Primers derived from the human sequence were used to detect the mRNAs corresponding to both Acs. For quantification of mRNAs we constructed internal standards for competitive quantitative reverse transcriptase PCR (RT-PCR). Both types of transcripts could be found in all investigated tissues except MNL where only type VI could be detected. Further we demonstrated a more than 60 times higher amount of AC V-mRNA in human heart compared to AC VI-mRNA. PMID- 10481933 TI - Stability of brain natriuretic peptide (BNP) in human blood samples. AB - Stability of immunoreactivity of human brain natriuretic peptide (BNP) in blood samples was investigated. After storage of the whole blood samples in the blood collecting tubes made of glass or polyethylene terephthalate (PET), residual immunoreactivity of BNP in the plasma was measured by sandwich radioimmunoassay for human BNP. BNP in the blood samples collected in the PET tubes were kept more stable than that in the glass tubes. The results suggested that commercially available PET tubes would enable more accurate BNP values and this would also help to simplify the sample preparation. PMID- 10481934 TI - A novel frameshift mutation in exon 6 (the site of Asn 291) of the lipoprotein lipase gene in type I hyperlipidemia. AB - A new heterozygous lipoprotein lipase gene defect has been identified in a type I hyperlipidemic patient at the position of notable amino acid Asn 291. The patient is a 33-year-old male. His body mass index (BMI) was 18.5 kg/m2. The total cholesterol (TC), triglycerides (TG) and high density lipoprotein-cholesterol (HDL-C) concentration from his fasting plasma were 4.8, 11.9 and 0.4 mmol/l, respectively. The lipoprotein lipase (LPL) activity and mass in the postheparin plasma (PHP) from the patient were 0.58 mmol/ml/h (normal range: 7.7+/-2.6) and 244 ng/ml (normal range: 192+/-30), respectively. The hepatic lipase activity of the PHP from the patient was 10.6 mmol/ml/h (normal range: 9.9+/-3.6). DNA analysis of the LPL gene revealed that this patient had a heterozygous one nucleotide deletion of A coding Asn 291, resulting in a premature termination of the LPL protein at amino acid residue 303. The other abnormality in the LPL gene of the proband was an amino acid residue 194 defect (Ile194-->Thr), which is known to cause a defective enzyme. A medium-chain triglyceride (MCT) loading test was conducted to find how this triglyceride affects plasma lipoprotein metabolism in this patient in a short term (Fig. 3). The plasma total cholesterol (TC) or high density lipoprotein (HDL)-C levels did not change significantly after oral administration of a fatty meal containing long chain triglycerides (LCT) or MCT. The plasma TG level, on the other hand, increased from 11.9 to 19.2 mmol/l (+61%) at 6 h after loading a fatty meal containing LCT, whereas the plasma TG levels tended to even decrease at 6 h after oral administration of an MCT, tricaprin (from 11.6 to 10.5 mmol/l (-9.4%)). These results suggest that MCT, as opposed to LCT, is useful for treatment of type I hyperlipidemia with a novel mutation at the notable amino acid Asn 291 of the LPL gene. PMID- 10481935 TI - A case of classical xanthinuria (type 1) with diabetes mellitus and Hashimoto's thyroiditis. AB - In a 62-year-old woman with non-insulin-dependent diabetes mellitus and Hashimoto's thyroiditis, hypouricemia was detected by a routine examination. Her plasma uric acid level was markedly low and urinary excretion of uric acid was undetectable. The high plasma and urine levels of xanthine were observed, although those of hypoxanthine were within normal ranges at rest after an overnight fast. After taking diet, plasma concentration and urinary excretion of hypoxanthine were markedly increased together with those of xanthine. The xanthine oxidase activity of duodenal mucosa was below the limits of detection. Allopurinol was metabolized to oxypurinol and pyrazinamide to 5 hydroxypyrazinamide in spite of no activity of xanthine oxidase, suggesting that aldehyde oxidase converted allopurinol to oxypurinol and pyrazinamide to 5 hydroxypyrazinamide. Based on these findings, she was diagnosed as having a subtype of classical xanthinuria type 1 with the normal plasma concentration of hypoxanthine in fast. PMID- 10481932 TI - Alpha-tocopherol acetate significantly suppressed the increase in heart interstitial 8-hydroxydeoxyguanosine following myocardial ischemia and reperfusion in anesthetized rats. AB - The effect of alpha-tocopherol acetate, an aqueous form of alpha-tocopherol, on the increase in heart interstitial 8-hydroxydeoxyguanosine (8-OH-dG) levels following myocardial ischemia/reperfusion was investigated. A microdialysis probe was implanted in the left ventricular interstitial space of anesthetized rat hearts. Myocardial ischemia was induced by ligating the left anterior descending coronary artery. Levels of 8-OH-dG in microdialysates were analyzed via an on line high-performance liquid chromatography system equipped with an electrochemical detector. The 8-OH-dG levels significantly increased (maximum 3.6 fold of increase relative to basal value) during the 60-min reperfusion stage following a 20 min ischemia. Administration of alpha-tocopherol acetate (20 mg/kg, intravenous, bolus) at 3 min prior to onset of reperfusion, significantly suppressed the reperfusion-induced increase in 8-OH-dG levels. These results suggested that one of the possible protective effect of alpha-tocopherol acetate was to reduce oxidative DNA damage during in myocardial ischemia and reperfusion. PMID- 10481936 TI - Acute hypermagnesemia and respiratory arrest following infusion of MgSO4 for tocolysis. AB - Hypermagnesemia (6.95 mmol/l) and respiratory arrest occurred to a 20-year-old female (G3P2002) at 26 weeks of gestation during tocolytic treatment with MgSO4.7H2O (density greater than plasmalyte) injected into an i.v. infusion bag containing 1 l of plasmalyte without mixing. The patient was rescued with calcium gluconate and normal pregnancy continued. It is important to adequately mix an i.v. solution after adding a drug particularly when the drug-containing solution has greater density than the parent i.v. solution. PMID- 10481937 TI - LD-IgG, IgA, IgM complex in a postburn patient. AB - A 4-year-old Chinese boy was admitted to our hospital with burns over 25% of his body. He had an extremely high LD activity (1743 U/l). LD isoenzyme analysis showed an abnormal band cathodic to LD-5. It was demonstrated by counter immunoelectrophoresis that the extra band was a complex of LD with IgG, IgA and IgM. Because LD-Ig complex is extremely rare in patients with burns, the clinical significance of this substance in blood remains unclarified. PMID- 10481938 TI - Regulation of BAX and BCL-2 expression in breast cancer cells by chemotherapy. AB - Optimizing chemotherapeutic drug delivery strategies relies, in part, on identification of the most clinically effective sequence, dose, and duration of drug exposure. The combination of dose intensive etoposide (VP-16) followed by cyclophosphamide has clinical efficacy in the treatment of advanced breast cancer. However, molecular mechanisms that underlie the effectiveness of this combination of chemotherapeutic agents have not been investigated. In this study we investigated regulation of BAX and BCL-2 expression by VP-16 and cyclophosphamide as a potential mechanism for the induction of breast cancer cell death induced by this regimen. There was a dose and time dependent increase in BAX expression in the breast cancer cell lines MCF-7, MDA-MB-435S, and MDA-MB A231 following in vitro treatment with 50-100 microM VP-16. Elevation of BAX protein expression in the presence of VP-16 alone did not correlate with reduced viability or induction of apoptosis in MCF-7, MDA-MB-435S, or MDA-MB-A231. VP-16 did effectively block the breast cancer cell lines evaluated (MCF-7 and MDA-MB 435S) at G2/M phase of the cell cycle, confirming activity of the drug in vitro. MCF-7 and MDA-MB-435S cells that were pre-treated with VP-16 and subsequently exposed to 1.0-12.0 microg/ml 4-hydroperoxycyclophosphamide (4HC), an active metabolite of cyclophosphamide, had markedly reduced viability when compared to matched controls treated with either VP-16 or 4HC individually. Consistent with this loss of viability, exposure of all three cell lines to the combination of VP 16 and 4HC resulted in higher BAX protein levels than those observed following treatment with either single agent. This combination of chemotherapeutic agents also resulted in reduced BCL-2 expression. These observations suggest that combination chemotherapy may derive its efficacy, in part, through coordinated regulation of specific gene products associated with apoptosis. Characterization of molecular events that underlie susceptibility of specific tumor cells to combination chemotherapeutic regimens may lead to additional improvements in treatment strategies for this disease. PMID- 10481939 TI - Successful co-treatment with LHRH-agonist for ovarian over-stimulation and cystic formation in premenopausal tamoxifen exposure. AB - The present study evaluates the potential beneficial effect of co-treatment with LHRH-agonist in resolving premenopausal tamoxifen's induced supraphysiological serum 17beta estradiol levels and persistent ovarian cysts. Ultrasonographic and serum hormonal evaluations were performed before, during, and following three consecutive injections of long acting LHRH-agonist administered to 14 premenopausal breast cancer patients treated with tamoxifen, who had supraphysiological serum 17beta estradiol levels and simultaneous persistent ovarian cysts. Within 3 weeks of the first LHRH-agonist injection, all patients had menopausal serum estradiol levels. Ovarian cysts completely disappeared within 2 months following the first injection. Following the discontinuation of LHRH-agonist co-treatment, serum estradiol levels remained in physiological levels and the ovaries remained a normal size in 64.3% of the patients for 13.3 +/- 11.5 months. 28.6% of the patients had a gradual reappearance of high serum estradiol levels and of ovarian cysts, and were, therefore, treated with a second course of LHRH-agonist. Following the second course, serum estradiol levels remained in physiological levels and the ovaries remained a normal size for 8-15 months. It is concluded that short duration of co-treatment with long acting LHRH agonist administered to premenopausal breast cancer patients treated with tamoxifen, successfully resolved the tamoxifen-induced supraphysiological serum 17beta estradiol levels and the ovarian cysts. PMID- 10481940 TI - Differentially expressed genes associated with the metastatic phenotype in breast cancer. AB - We have previously shown that human breast carcinoma cells demonstrating an interconverted phenotype, where keratin (epithelial marker) and vimentin (mesenchymal marker) intermediate filaments are both expressed, have an increased ability to invade a basement membrane matrix in vitro. This increase in invasive potential has been demonstrated in MDA-MB-231 cells, which constitutively express keratins and vimentin, and in MCF-7 cells transfected with the mouse vimentin gene (MoVi). However, vimentin expression alone is not sufficient to confer the complete metastatic phenotype in MoVi cells, as determined by orthotopic administration. Thus, in the present study, differential display analysis was utilized to identify genes that are associated with the invasive and/or metastatic phenotype of several human breast cancer cell lines. Forty-four of 84 PCR fragments were differentially expressed as assessed by Northern hybridization analysis of RNA isolated from MCF-7, MoVi, and MB-231 cell lines. Polyadenylated RNA from a panel of poorly invasive, invasive/non-metastatic, and invasive/metastatic breast carcinoma cell lines was used to differentiate between cell-specific gene expression and genes associated with the invasive and/or metastatic phenotype(s). We observed that lysyl oxidase and a zinc finger transcription factor were expressed only in the invasive and/or metastatic cell lines; whereas, a thiol-specific antioxidant and a heterochromatin protein were down-regulated in these cells. In contrast, tissue factor was expressed only in breast carcinoma cell lines having the highest invasive potential. These results suggest that specific genes involved in breast cancer invasion and metastasis can be separated by differential display methodology to elucidate the molecular basis of tumor cell progression. PMID- 10481941 TI - Cathepsin D expression by cancer and stromal cells in breast cancer: an immunohistochemical study of 1348 cases. AB - This study was aimed at investigating the influence of cathepsin D (CD) expression by cancer cells and stromal cells on breast cancer prognosis. This is a study of 1348 node-positive (NPBC) and node-negative (NNBC) breast cancers diagnosed between 1980 and 1986 and with a minimum follow-up of 5.2 years. CD expression was assessed by immunohistochemistry on archival material using a polyclonal antibody. The expression by cancer and stromal cells was assessed separately and correlated with distant metastasis free (DMFS) and overall survival (OS). Cancer cells expressed CD (more than 10% cells expressing CD) in 38.9% of cases and reactive stromal cells in 43.6%. CD expression by reactive stromal cells, and not cancer cells, correlated with several factors of poor prognosis by cancer cells. A strong association was also found with expression of other proteases (stromelysin-3, gelatinase A, and urokinase Plasminogen Activator) by these same reactive stromal cells. CD expression by cancer cells did not predict DMFS or OS but, by univariate analysis, CD expression by reactive stromal cells was associated with earlier recurrence and shorter survival in NNBC (p = 0.0425) and NPBC patients submitted to adjuvant chemotherapy (p = 0.0234). However, CD expression by reactive stromal cells remained a significant predictor of recurrence by multivariate analyses only in a subgroup of NPBC submitted to adjuvant chemotherapy. Overall, those data support the concept that proteases produced by reactive stromal cells are under cancer cell stimulation and that CD by stromal cells, and not cancer cells, influences the prognosis, but only in a subgroup of patients with breast cancer. PMID- 10481942 TI - Synergistic action of apoptosis induced by eicosapentaenoic acid and TNP-470 on human breast cancer cells. AB - The effects of eicosapentaenoic acid (EPA) and an angiogenesis inhibitor (TNP 470) on the suppression of breast cancer cell growth were examined in five human breast cancer cell lines (MDA-MB-231, T-47D, MCF-7, KPL-1, and MKL-F). In all five cell lines, EPA and TNP-470 alone both showed tumor growth inhibition in a time- and dose-dependent manner, and in combination, a synergistic effect was seen at high concentrations. EPA plus TNP-470 treatment evoked apoptosis as confirmed by the appearance of sub G1 populations, by DNA fragmentation, and by cell morphology. With the combination, the expression of Bax and Bcl-xS, the apoptosis-enhancing proteins, was more up-regulated and that of Bcl-2 and Bcl-xL, the apoptosis-suppressing proteins, was more down-regulated compared to the use of EPA or TNP-470 alone, suggesting that their synergistic effect was due to an acceleration of apoptosis. PMID- 10481943 TI - Reduced expression of p27Kip1 protein is associated with poor clinical outcome of breast cancer patients treated with systemic chemotherapy and is linked to cell proliferation and differentiation. AB - The cyclin-dependent kinase inhibitor p27Kip1 is a negative regulator of cell proliferation. Its expression is known to be altered in a proteasome-dependent manner without changes in DNA level. Reduced expression of p27Kip1 is associated with aggressive behavior in a variety of human cancers. We investigated expression of p27Kip1 protein in human breast cancer using immunohistochemistry to assess its biologic implication along with cell-cycle analysis by flow cytometry. A total of 68 patients with invasive ductal cancer received adjuvant chemotherapy with cyclophosphamide, methotrexate, and 5-FU every 3 weeks for six cycles. In epithelial cells of normal and benign breast disease, expression of p27Kip1 was well preserved while its expression markedly decreased in breast cancer (45 of 68). Expression of p27Kip1 is significantly reduced in poorly differentiated cancers and in the advanced stage of the disease. Levels of p27Kip1 expression correlated with cell populations in G0/G1 phase of the cell cycle. In survival analysis, p27Kip1 was useful to predict disease free survival but not overall survival of the patients after adjuvant chemotherapy. In summary, p27Kip1 seems to have a role in the cell proliferation and differentiation process during carcinogenesis of breast cancer. The results of the present study suggest that p27Kip1 can be used in predicting response to systemic chemotherapy in a subset of patients with breast cancer. PMID- 10481944 TI - Pathobiologic identification of two distinct breast carcinoma subsets with diverging clinical behaviors. AB - Many different pathological and biological variables which characterize breast carcinomas have been found to be associated. The aim of this work was to analyze the complex relationship among these parameters. The pathologic, biologic, and clinical characteristics of a series of primary breast carcinomas from 676 patients were retrospectively investigated. Multiple correspondence analysis of 13 factors revealed clustering of eight pathobiologic variables, that is histologic grade, necrosis, lymphoid infiltration, number of mitoses, c-erbB-2 overexpression, p53, progesterone receptor, and bcl2 expression. An index for each tumor calculated on the basis of these eight factors served to distinguish two different tumor phenotypes, designated A and B. Phenotype A is represented by tumors sharing most of the biologic features of normal breast tissues: indeed, these tumors are characterized by a relatively high degree of differentiation, low proliferation, no necrosis or leukocyte infiltration, and no gene alterations. By contrast, phenotype B is quite divergent from the normal tissue because of its poor differentiation, high proliferation, frequent gene alterations and evidence of a host immune reaction. As regards the disease progression, these two subsets showed marked differences: phenotype A tumors had a low recurrence rate per year that remained constant over time and affected more frequently elderly patients, whereas group B tumors showed high aggressivity in the first years after surgery followed by a low long-term recurrence rate and were more frequently seen in younger patients. These data suggest that breast carcinoma consists of two different subsets that can be identified on the basis of pathobiologic features. PMID- 10481945 TI - Down-regulation of gelsolin expression in human breast ductal carcinoma in situ with and without invasion. AB - Expression of gelsolin, an actin filament regulatory protein, in human breast ductal carcinoma in situ (DCIS) was analyzed by immunohistochemistry using a monoclonal antibody. Formalin-fixed paraffin-embedded tissues from 59 pure DCIS specimens and 33 DCIS specimens with associated invasive components were evaluated for gelsolin reactivity and compared to eight normal breast cases and 76 invasive breast cancers. The proportion of cases exhibiting negative/low expression of gelsolin in the epithelium was as follows -- normal, 0%; pure DCIS, 56%; DCIS associated with invasion, 58% in the DCIS component and 66% in the invasive component; invasive carcinoma, 70%. These data demonstrate that down regulation of gelsolin expression in breast epithelium frequently parallels progression to malignancy. Testing gelsolin expression (normal vs. negative/low levels) in the DCIS lesions for associations with patient age or any of the following histopathologic parameters revealed no significant (95% probability level) correlations -- tumor size; pathologic (Van Nuys system) grade; nuclear grade; necrosis; presence of histologic calcifications; presence or type of adjacent benign lesions; architectural histologic pattern; and mammographic extent. Gelsolin loss was more commonly associated with mammographic soft tissue lesions as compared to calcified lesions (P = 0.009). A positive trend of borderline significance (P = 0.06) found in the DCIS with invasion group was a correlation between down-regulated gelsolin expression in the DCIS component and size (< versus > or = 15 mm) of the invasive tumor. In conclusion, reduced gelsolin protein is detectable in at least half of breast lesions which have progressed to DCIS. The trend between increasing gelsolin loss and malignant progression from normal epithelium to DCIS to invasive breast cancer (P < 0.0001) suggests additional investigation is needed to determine the potential of altered gelsolin expression as a marker for prognosis and for therapeutic interventions in breast cancer. PMID- 10481946 TI - Assessment of quality of life in women undergoing hormonal therapy for breast cancer: validation of an endocrine symptom subscale for the FACT-B. AB - Existing quality of life instruments do not include adequate items to measure the side effects and putative benefits of hormonal treatments given in breast cancer. We report the development and validation of an 18 item endocrine subscale (ES) to accompany a standardised breast cancer quality of life measure, the Functional Assessment of Cancer Therapy (FACT-B). The FACT-ES (FACT-B plus ES) was tested initially on 268 women with breast cancer receiving endocrine treatments. Alpha coefficients for all subscales demonstrated good internal consistency (range alpha = 0.65-0.87). Test-retest reliability of the ES indicated good stability (r = 0.93, p < 0.001). Advanced breast cancer patients' quality of life was high, showing the efficacy of endocrine therapy, but women with primary disease reported better physical, social, and functional well-being and fewer breast cancer concerns. Most frequently reported symptoms were loss of sexual interest (31%), weight gain (25%), and hot flushes (24%). Significant differences were found between treatment groups for hot flushes and vaginal dryness. Two assessments of the instrument's responsiveness to change were made; 32 women in a clinical trial of endocrine therapy and 18 women without breast cancer taking HRT completed the FACT-ES at baseline, 4, 8, and 12 weeks. Trial patients reported significantly more symptoms at 8 and 12 weeks than at baseline. Women taking HRT reported significantly fewer or less severe symptoms than at baseline. In conclusion the FACT-ES has acceptable validity and reliability and is sensitive to clinically significant change, making it suitable for clinical trials of endocrine therapy. PMID- 10481947 TI - Capillary electrophoresis of peptides and proteins in acidic, isoelectric buffers: recent developments. AB - The use of isoelectric buffers in capillary zone electrophoresis is here reviewed. Such buffers allow delivery of very high voltage gradients (up to 1000 V/cm in relatively large bore capillaries, e.g. 75-100 microm I.D.), permitting separations of the order of a few minutes and thus conserving (in fact favouring) very high resolution due to minimal, diffusion-driven, peak spreading. Isoelectric Asp (pI 2.77 at 50 mM concentration and 25 degrees C) provides a medium of high resolving power for generating peptide maps. In difficult cases, of coincident titration curves, the pH can be moved up to higher values (e.g. pH 3.0 for 30 mM Asp) thus eliciting separation of unresolved peptides at pH 2.77. This was illustrated by running peptide maps of tryptic digests of human beta globin chains. Also imino diacetic acid (pI 2.33 at 50 mM concentration) allows generation of high resolution peptide maps. Isoelectric Asp, in presence of 7 M urea and 0.5% hydroxyethyl cellulose (Mn = 27 000 Da) is also the preferred medium for fast separation and analysis of storage proteins in cereals, such as gliadins in soft and durum wheat and zeins in maize. A solution of 50 mM iminodiacetic acid (pI 2.23) containing 7 M urea and 0.5% hydroxyethylcellulose (apparent pH 3.2) is effectively used as background electrolyte for fast separation of heme-free, denatured globin (alpha and beta) chains. In the presence of neutral to neutral amino acid substitutions, it is additionally shown that the inclusion of 3% surfactant (Tween 20) in the sample and background electrolyte induces the separation of the wild-type and mutant chains, probably by a mechanism of hydrophobic interaction of the more hydrophobic mutant with the detergent micelle, via a mechanism similar to 'micellar electrokinetic chromatography'. PMID- 10481948 TI - Using affinity capillary electrophoresis to study the interaction of the extracellular binding domain of erythropoietin receptor with peptides. AB - We have shown that affinity capillary electrophoresis (ACE) can be utilized to screen peptides that bind to the extracellular binding domain of the erythropoietin receptor (EBP). The comparison of the cyclic peptides GGTYSCHFGPLTWVCKPQGG (EMP1) GGTYSCHFGPLTAVCKPQGG (EMP13), and LGRKYSCHFGPLTWVCQPAKKD (EMP37) with the linear peptides HFGPLTWV (EMP26) and FMRF as ACE buffer additives were investigated. When EMP1 and EMP37 were the buffer additives, an abrupt change in the electrophoretic mobility of EBP was observed in the electropherogram. When EMP13, EMP26, and FMRF were examined under identical ACE conditions as EMP1 and EMP37, no significant change in the electrophoretic mobility of EBP was observed. These results correlate well with previously reported IC50 competitive binding data; that is, EMP1 and EMP37 bind to EBP while EMP13 and EMP26 bind very weakly. These observations strongly infer that peptide.EBP dimerization were induced by EMP1, and EMP37 but not by EMP13, EMP26 or FMRF. This ACE method provides a rapid tool for the detection of small peptides or drugs that bind to EBP. PMID- 10481949 TI - Voltammetric study of charge transfer across supported bilayer lipid membranes (s BLMs). AB - Stable bilayer lipid membranes on a metal support (s-BLMs) and on an agar gel salt bridge (sb-BLMs) have been formed and their potential usefulness as practical sensors demonstrated. This paper presents the preparation method of s BLMs and sb-BLMs, and the application of cyclic voltammetry (CV) to their investigation. The instrument and data analysis of the CV are described. The application of CV to the C60-doped BLM system is presented. This technique is a basis for biosensor development, and a useful tool for membrane research. PMID- 10481950 TI - Stabilization of alginate beads using radiation polymerized polyacrylamide. AB - A technique has been described for the stabilization of calcium alginate beads using radiation polymerized acrylamide. The technique involved dropping a mixture containing the cells (20%), sodium alginate (2%), acrylamide (2.5%) and N-N' methylene-bis-acrylamide (0.1%) through a syringe needle into cold (-75 degrees C) toluene. The frozen beads obtained were exposed to 60Co gamma-rays (0.5 KGy) and were then thawed in 0.1 M CaCl2 solution. Unlike the calcium alginate beads the conjugate beads were not found to be dissolved when incubated in 3% trisodium citrate solution. Stabilized beads containing entrapped yeast cells could be reused for over 15 batches for the inversion of sucrose without loss in activity or chemical integrity of the beads. PMID- 10481951 TI - Overcoming false negatives due to the genomic context in polymerase chain reaction amplification. AB - In some instances defined genomic regions are so poorly amplified that they seem to be unamplifiable. A protocol was developed which allows good PCR amplifications by the use of restriction digestion combined with the elution of a pool of restriction fragments of defined size range from agarose gel after electrophoresis. We describe the application of the method in the PCR amplification of a region of the white locus of Drosophila melanogaster that otherwise may be considered as a negative result. PMID- 10481952 TI - Detection and assay of proteases using calf lens beta-crystallin aggregate as substrate. AB - The eye lens protein, betaL-crystallin, aggregates and yields a turbid solution upon refolding from its denatured state. We have observed that the addition of trace amounts of protease results in clearing of this turbidity. Based on this observation, we have developed a simple and rapid method for the detection and assay of proteases. This assay can be performed in the pH range of 6.0-9.0. We could assay the activity of trypsin at a concentration as low as 5 microg/ml. PMID- 10481953 TI - Contaminant eluted from solid-phase plasmid affinity-purification protocol columns is not found using liquid-phase methods and can be prevented. AB - The preparation of high quality plasmid DNA is a necessary requirement for most molecular biology applications. We compared four different large plasmid preparation protocols, which were based on either a liquid-phase approach (Triton lysis) or purification of alkaline lysis bacterial extracts followed by supercoiled plasmid purification on affinity columns. Two host Escherichia coli strains, JM 109 and INValphaF', were used to grow the test plasmids for comparison of product plasmid DNA produced from the four different plasmid isolation methods. While the DNA grown in E. coli strain JM109, prepared by liquid-phase Triton lysis was appropriately restricted by 12 restriction enzymes, this was not the case for any of the JM109-grown DNA purified by any of the affinity column solid-phase approaches. In contrast to this, when the plasmid DNA was grown in E. coli strain INValphaF', most restriction enzymes cut DNA appropriately, irregardless of the plasmid preparation protocol used. It seems that an impurity commonly eluted with the DNA from all three of the solid-phase DNA columns had an equal effect on the above enzymes using the common host strain JM109, but not strain INValphaF'. PMID- 10481954 TI - Histopathological and functional effects of radiation therapy in obstructive uropathy. AB - Leukocyte influx into the cortex and medulla of the kidney is seen following acute ureteral obstruction. To investigate the macrophage influx and alterations in glomerular filtration rate (GFR) and to evaluate the effects of early radiation therapy on these parameters, we evaluated 20 rabbits in four groups. In the first group preoperative technetium-99m scanning preceding bilateral ureteral complete obstruction was performed. All rabbits received whole body irradiation of 1316 rads. Then the bilateral ureteral obstruction was released after 24 hours. Two hours after releasing, the last scanning was performed. The kidneys were immediately removed and histopathological examination was done. In the second group, all procedures except radiotherapy were done. The third group underwent sham laparotomy to assess the effects of surgical trauma on renal functions. The fourth group of rabbits was considered as control. We have observed an apparent macrophage influx into the cortex and medulla of the kidney following bilateral ureteral obstruction (p=0.000). However, irradiation reduced the infiltration significantly (p=0.000). Also in cases who received radiotherapy GFR was preserved to a significantly higher degree (p=0.000). We concluded that irradiation following acute ureteral obstruction has protective effects on renal function through abolition of the infiltrating cells. PMID- 10481955 TI - Prognostic index of renal carcinomas. AB - Authors analysed the survival of 126 patients operated for kidney carcinomas. The data on anamnesis, laboratory results and TNM histological classification were compared with the results on "relative survival" and correlation analysis was performed. Based on these, the 12 most important criteria (with the highest correlation coefficient) were emphasized and regression equation was formed based on the TSP (Time Series Processing) Version 4 (Hall). Using this method, a numerical prognostic index can be established for the judgement of prognosis regarding certain patients. PMID- 10481956 TI - Radiopaque 2,8-dihydroxyadenine lithiasis. AB - 2,8-Dihydroxyadenine (DHA) lithiasis is a rare type of urinary stone disease and the deficiency of adenine phosphoribosyltransferase (APRT) activity is known to be the cause of this disease. To identify a 2,8-DHA stone is important because this stone is well managed by medical therapy. Regarding a radiological finding, previous reports have considered 2,8-DHA to be radiolucent like uric acid stone. This report is a case of radiopaque 2,8-DHA stone and proposes that the composition of urinary stones should always be investigated for adequate medical treatment. PMID- 10481957 TI - Acute post streptococcal interstitial nephritis in an adult and review of the literature. AB - While glomerular lesions are a recognized sequel to infection with group A beta haemolytic streptococci, literature on primary tubular lesions, as remote effects of streptococcal infection, is scanty. A case of interstitial nephritis in a 29 year-old woman following streptococcal sore throat is described. This adult admitted with tonsillitis developed acute renal failure from acute interstitial nephritis and subsequently had full recovery of renal function. Acute interstitial nephritis should be included in the differential diagnosis of patients with streptococcal infections who develop acute renal failure. PMID- 10481959 TI - Concomitant renal and prostatic aspergillosis. AB - Aspergillosis limited to the urinary tract is a rare disease seen most often in patients with altered immune status. Only 19 cases of renal aspergillosis including 3 with AIDS and 4 cases of isolated prostatic aspergillosis have been reported. We report the first case of concomitant renal and prostatic aspergillosis in a non-immunocompromised patient who presented with pyrexia of unknown origin and with dysuria. The diagnosis was based on the demonstration of characteristic hyphal elements on direct microscopy and isolation of the fungus in the culture of pus from the kidney. In view of obstructive prostatic enlargement and left non-functioning renal mass, transurethral resection of the prostate and left nephrectomy were performed in a single session with successful outcome. The aetiopathogenesis and brief review of the literature are discussed. PMID- 10481960 TI - Ultrasonographic urinary tract abnormalities in Schistosoma haematobium infection. AB - OBJECTIVE: Evaluation of ultrasonographic abnormalities with active Schistosoma haematobium infection in Yemeni patients. METHODS: As part of a cooperation between a private hospital and Schistosomiasis Control Project in Yemen, laboratory and ultrasonographic examinations were performed in 158 patients (8 females, 150 males, mean age: 17 years) with active Schistosoma haematobium infection. Upper urinary tract dilation, lower ureter changes and bladder wall abnormalities (thickness, hyperechogenicity and polypoid lesions) were registered. Laboratory findings and ultrasonographic changes were evaluated and assorted according to age groups of the patients. RESULTS: Twenty-eight patients (18%) showed no ultrasonographic morphological lesion. Bladder abnormalities (thickness, hyperechogenicity and polypoid lesions) were found in 130 patients (82%) and upper tract lesions in 86 patients (54%). No upper tract lesions were seen without bladder abnormality. Polyps within the lower ureteric lumen occurred in four patients. In patients with polypoid lesions, higher incidence of severe infection was found. One patient had urinary bladder mass. CONCLUSION: Urinary schistosomiasis has typical sonographic features, however, it may occur also without ultrasonographic morphological lesion. Upper tract lesions seem to develop only with lower tract abnormalities. PMID- 10481958 TI - A new experimental model for partial ureteric obstruction in sheep. AB - This study aimed to develop a new experimental model of partial ureteric obstruction in sheep. Graded obstruction of the ureter using various sized catheters (Fr 3-8) for variable durations up to 6 weeks was performed in 19 sheep. The kidneys were studied by radionuclide diuretic renography for up to 21 weeks and had histopathological examination at the end of the experiment. Catheters of Fr 3 or Fr 5 should be used to block the ureter for a minimum of three weeks to produce sufficient functional and pathological changes to be studied. The advantages and disadvantages of this model are discussed. PMID- 10481961 TI - Paediatric nosocomial urinary tract infection at a regional hospital. AB - From a total of 26,603 admissions to the paediatric wards, 1360 paediatric nosocomial urinary tract infections (PNUTI) were identified during a 5-year retrospective chart review at the SFGH. The ages ranged from 3 days to 13 years, with 46% boys and 54% girls. The highest rates of PNUTI per service per 100 admissions were seen in the nursery (11.28) followed by paediatric surgery (2.89) and paediatric medicine (2.86). Although the greatest number of PNUTI occurred in the nursery, comparison between the years was not statistically significant. About 90% (1218 of 1360) of PNUTI occurred in catheterized patients. No documentation was found specifying the type of catheterization (intermittent or continuous). About 90% (1210 of 1360) of isolates were single organisms with Escherichia coli, Proteus mirabilis, Klebsiella spp. and Group B streptococci accounting for a total of approximately 70% of all pathogens. However, the composition of the most common isolate in each service differed. The most common isolate in the nursery was E. coli, in the paediatric medical and surgical services the most common isolates were Klebsiella spp. and Proteus mirabilis, respectively. Proteus mirabilis was isolated predominantly from boys with structural abnormality of the urethral tract. No PNUTIs were complicated by bacteraemia. The antibiotics with least effectiveness (in increasing order) for UTIs were cephalexin, ampicillin, trimethoprim, co-trimoxazole and tetracycline. The most effective antibiotics were nalidixic acid, gentamicin and amoxicillin clavulanic acid. PMID- 10481962 TI - In vitro effect of amezinium on the rabbit urinary bladder: muscle strip and whole bladder evaluation. AB - Dose-response studies for norepinephrine and amezinium were obtained for isolated muscle strips and in vitro whole bladder preparations to investigate the effect of amezinium on the bladder. Specimens (strips from the bladder base and body, and the whole bladder) were obtained from mature female rabbits. By the muscle strip study, amezinium showed a weak contractile effect (up to 40% of spontaneous contraction) on strips from the bladder base. Muscle strips from the bladder body presented no response to amezinium. In the whole bladder preparation, administration of norepinephrine showed an increase in intravesical pressure in 4 out of 7 rabbits, whereas administration of amezinium had negligible effect on intravesical pressure. Administration of amezinium showed no influence on strips from the bladder body and the whole bladder preparation. Direct contractile response in muscle strips from the bladder base by amezinium appeared to be a favourable effect considering urinary storage. PMID- 10481964 TI - Aetiological factors of bladder cancer in the Aegean region of Turkey between the years 1985-1996. AB - A great majority of urological cases are bladder tumours. The purpose of this study is to bring out the aetiological factors related to bladder tumours. The parameters such as age, sex, profession, age at tumour occurrence, smoking, drinking habits, such as the level of consumption of tea and coffee, and accompanying urological diseases were evaluated. Three hundred and forty-seven patients with bladder tumours were included in this study. Of them 332 (95.6%) were males and 15 (4.4%) females. The average age was 62.1 (22-87) years. Of the patients 326 (93.9%) smoked, 175 (50.4%) lived in cities and the other 49.6% lived in the countryside. Of the tumours 89.9% were transitional cell carcinomas. In conclusion, bladder tumours are closely related to consumption of tobacco factors and profession. The risk of tumour development increases progressively in people who are exposed to industrial agents and agricultural chemicals. PMID- 10481963 TI - Use of diagnostic categories in urinary cytology in comparison with the bladder tumour antigen (BTA) test in bladder cancer patients. AB - In recent years the use of diagnostic categories for extragenital cytology has increasingly been discussed as an approach to improve the quality of reports. Diagnostic categories reflect the adequacy of the materials for interpretation and the presence or absence of cancer cells. There is a tendency to add intermediate groups as qualifying probably malignant cases or findings associated with a serious cancer risk. Since 1971 we have added one of the following to the final diagnosis in all cases: unsatisfactory for cytological diagnosis, negative for cancer, repeat test suggested, suspicious of cancer, and positive for cancer. To evaluate whether diagnostic categories are useful for comparison of cytological results with those of an alternative test, cytological data were compared with the results of the Bard bladder tumour antigen (BTA) test in voided urine from 119 patients (76 with and 43 without bladder cancer). The diagnostic categories enabled us to calculate sensitivities and specificities of cytology based on different thresholds or decision levels. The BTA test had significantly higher sensitivity (79%) and lower specificity (60%) than urinary cytology with three different thresholds in cytology results (sensitivities: 16-43%, specificities: 81-100%). The present findings suggest that diagnostic categories improve comparison of cytologic results with those of alternative screening and diagnostic aids such as the BTA test. PMID- 10481966 TI - Comparison of voiding disturbances in patients with LUTS and BPH at home and by episodic checks at an outpatient clinic. AB - To clarify the presence of voiding disturbances in patients with lower urinary tract symptoms (LUTS) and benign prostatic hyperplasia (BPH), measurements of the capacity at normal desire to void (CNDV), residual urine volume (RUV) and RUV rate at home and in a hospital were done. It has been found that (1) RUV and RUV rate in patients with BPH were markedly greater than that in the normal group and LUTS patients staying at home. Moreover, there was no difference in RUV and RUV rate between the normal group and the LUTS patients. (2) RUV and RUV rate in patients with LUTS and BPH decreased more markedly without changing CNDV when the patients stayed at home than when they underwent episodic checks in a hospital. In conclusion, frequent checks of LUTS and BPH in patients in their domestic environment can probably be utilized for management or determination of treatment. PMID- 10481965 TI - Penetration of a single infusion of ampicillin and sulbactam into prostatic tissue during transurethral prostatectomy. AB - Antibiotic prophylaxis is recommended in endoscopic urological operations to diminish the rate of intraoperative septic complications and relevant urogenital infections. The objective of the study was to determine the tissue concentrations of preoperatively administered ampicillin and sulbactam in the human prostate in patients undergoing transurethral resection (TUR-P) for benign prostatic hyperplasia (BPH). In 19 patients (mean age 68.7 years) the serum and tissue concentrations of ampicillin and sulbactam were determined. For all patients the dosage was administered as a single infusion over 15 min at a 2:1 ratio, i.e. 2 g ampicillin and 1 g sulbactam. The serum and prostatic tissue samples were taken 15 to 55 min (mean 29.5 min) after infusion. Of transurethral resected prostatic chips 3 g were immediately collected after resection and stored at -70 degrees C. Ampicillin was determined by bioassay and sulbactam was determined by gas chromatography/mass spectrometry. Tissue concentrations of ampicillin ranged from 0.42 to 548.33 mg/kg (median 47 mg/kg). Tissue concentrations of sulbactam ranged from 0.15 to 249.74 mg/kg (median 19 mg/kg). Six (32%) of 19 patients showed a tissue concentration of ampicillin <4 mg/kg (MIC90), respectively 5 (26%) patients of sulbactam <8 mg/kg. The mean serum concentrations at tissue sampling time were 118.8+/-48.9 mg/l respectively 32.2+/-12.2 mg/l. There exists a high variability of intraprostatic concentrations of ampicillin and sulbactam after single infusion in patients with BPH. In a relevant part of patients both compounds do not exceed the minimal inhibitory concentrations (MIC) of important bacterial pathogens. A single shot infusion of 3 g ampicillin/sulbactam for intraoperative antibiotic prophylaxis is not sufficient in patients undergoing TUR-P. It seems that the prostate cannot be compared to other tissues in view of penetration of ampicillin/sulbactam. PMID- 10481967 TI - Serum PSA and age-specific reference ranges in patients with prostatism symptoms. AB - Prostatic specific antigen (PSA), a tumour marker helpful in the diagnosis and follow-up of prostate cancer, may rise due to causes other than prostate cancer (i.e. BPH, acute prostatitis, etc.). Investigations in order to increase the sensitivity and specificity of PSA in prostate carcinoma are being carried out. Serum PSA levels of patients with prostatism with regard to age as well as these levels in the male population at risk but without clinical prostatic disease (those above the age of 40) should be well documented. The aim of this study is to find age-specific values and ranges of PSA in patients with prostatism symptoms. PMID- 10481968 TI - The prognosis of stage A patients treated with the antiandrogen chlormadinone acetate. AB - To determine the efficacy of the antiandrogen chlormadinone acetate as a treatment option for stage A prostate cancer, a retrospective analysis of 111 patients who received chlormadinone acetate was done. Of 55 stage A1 patients, progression was seen in one patient (1.8%) 8 years after the diagnosis. Six out of 56 stage A2 patients (10.7%) showed disease progression at 1-7 years (mean 68 months) after the diagnosis. Progression rates associated with antiandrogen therapy for stage A1 and stage A2 patients were lower than those without treatment reported in the literature. It could be assumed that antiandrogen treatment with chlormadinone acetate inhibited the progression. PMID- 10481969 TI - The role of autosomal cell apoptosis regulator genes in human spermatogenesis. AB - Immunohistochemical analysis of the expression of the autosomal apoptosis regulator genes, BAX, bcl-2, p53, and cyclic-AMP responsive element modulator (CREM) in testis biopsies from infertile men demonstrated that BAX, bcl-2, and p53 immunoreactivity was absent irrespective of seminiferous tubule histology. In contrast, cell-specific CREM immunoreactivity in round spermatids, with complete absence of CREM in patient biopsies showing spermatocyte maturation arrest and Sertoli cell only, was evident, suggesting a possible role of autosomal genes in the regulation of apoptosis in human spermatogenesis regulation. PMID- 10481970 TI - Testicular torsion: can the testicle be saved one week later? AB - An unusual case of a 15-year-old male who presented with a characteristic clinical syndrome of testicular torsion one week prior to our observation, is reported. During our intervention the testicle was not removed but, on the contrary, restoration of a satisfactory testicular albugineal appearance was achieved. PMID- 10481971 TI - Diphallus with urethral duplications. AB - We report a case of glandular diphallus with incomplete urethral duplication associated with rotation anomaly in the right kidney, complete ureteral duplication and ectopic ureteral orifice in the left. The bladder was single with good sphincter control. At operation, the hypoplastic glans was resected and the urethra opening into this glans was anastomosed side by side with the other urethra. Diphallus and incomplete urethral duplication are discussed in the light of evidence in the literature. PMID- 10481972 TI - Patient-partner satisfaction with intracavernous medication supported with oral agents in selected cases of Peyronie's disease. A ten-month follow-up study. AB - We attempted to find the most adequate treatment option for some selected cases of Peyronie's disease. Between 1993 and 1996, 38 patients with Peyronie's disease and erectile dysfunction were treated with intracavernous medication, supported with oral colchicine and vitamin E. None of the cases had severe angulation of penis and intolerable pain during erection. Quality of sexual life was assessed by CWRU questionnaire. After ten-month follow-up we found improvement in all parameters, in CWRU. The symptoms of 24 cases diminished. The 21 partners examined were also satisfied with the therapy. We conclude that intracavernous medication combined with oral agents is a useful alternative treatment in selected cases of Peyronie's disease. PMID- 10481974 TI - Prediction of protein retention at gradient elution conditions in ion-exchange chromatography. AB - This work presents a prediction procedure for protein retention in ion-exchange chromatography, where two linear gradient experiments of different length give the protein retention time at other linear gradients. The procedure predicts the retention time of early and late eluting proteins with similar precision and predictions by extrapolation deviate approximately 3% or less from the experimental retention times. By using the ionic strength, this procedure predicts protein retention times obtained with divalent ions in the eluent more accurately than a well-established procedure that uses the protein co-ion concentration. PMID- 10481973 TI - Lipid and lipoprotein ratios as risk factors of atherosclerosis in patients with chronic renal insufficiency (CRI). AB - The serum levels of lipids, apolipoproteins and lipoprotein ratios in 19 healthy persons and 20 patients with uraemia not dialyzed were determined. Based on creatinine level the patients were divided into two groups: L (serum creatinine 2 5 mg/dl) and H (serum creatinine 5-10 mg/dl). Dyslipoproteinaemia in uraemic patients is already manifested in the early stages of the disease through its abnormal apolipoproteins rather than lipid profile and it suggests a high risk of atherosclerosis. PMID- 10481975 TI - Generalised H-point standard addition method for the isolation of the analyte signal from the sample signal when coelution of unknown compounds occurs in liquid chromatography. AB - The generalised H-point standard addition method (GHPSAM) is proposed for isolating the analytical signal of an analyte from the signal of an unknown sample. Samples containing two and three coeluting compounds have been analysed. The accuracy of the predictions depends on the shape of the analyte and interferent spectra but not on the degree of chromatographic overlap. This methodology involves the location of linear intervals for the unknown interference spectrum from the spectrum of the sample. Once the linear interval has been found the selection of three wavelengths within the interval will allow the cancellation of the signal of the unknown interferent. The method has been applied to the determination of diuretics, amphetamines and phenols in water. PMID- 10481976 TI - Equation for calculating surface coverage from end-capping of chromatographic bonded phases. AB - An equation was derived to calculate the surface density of trimethylsilyl groups (alphaTMS) on bonded chromatographic stationary phases that have undergone primary as well as secondary ('end-capping') chemical modification. The new equation is an extension of that published by Berendsen-de Galan for calculating primary surface coverage and, likewise, alphaTMS is calculated in terms of the carbon content (% by weight) of the bonded material before and after end-capping, specific surface area of the starting silica and structural information (molecular weight and number of carbon atoms) of the anchored groups. The new equation is valuable when a thorough characterization of bonded stationary phases is desirable and, if used along with the Berendsen-de Galan equation, it affords total ligand coverage information. Application of the new equation to correct for measurable carbon content of the starting support leads to a more accurate expression for surface coverage from primary as well as secondary bonding. The scope and limitations of the new equation are discussed. PMID- 10481977 TI - Empirical relationship between chiral selectivity and mobile phase modifier properties. AB - An empirical relationship was derived which relates properties of the mobile phase modifier to the chiral selectivity factor for a given analyte/chiral selector combination. Using carbon dioxide and heptane-based mobile phases, the effect of various mobile phase modifiers on Pirkle-type stationary phases may be accurately modeled using a two-parameter equation. Similar results are obtained using cellulosic stationary phases with carbon dioxide-based mobile phases. Modeling separations performed using heptane-based mobile phases with cellulosic stationary phases were not successful. The predictive ability of this modeling approach was demonstrated using novel modifiers and chiral analytes. PMID- 10481978 TI - Automated on-line membrane extraction liquid chromatographic determination of phenols in crude oils, gasolines and diesel fuels. AB - Determination of phenols in crude oils and derived fuels requires a sample pretreatment step, usually performed by liquid-liquid extraction or preparative chromatography. In this work, sample preparation is accomplished using a silicone membrane separation unit coupled on-line to a high performance liquid chromatograph with amperometric and ultraviolet detection. The contents of phenol, cresols and dimethylphenols were determined in thirty three samples including three crude oils, twenty gasolines and ten diesel fuels. The whole set up is fully automated through a feed-back system that allows the microcomputer controlling the process to examine the signals in real time and to make decisions while the experiment is running. PMID- 10481979 TI - Isolation and characterization of cyclic alpha-(1-->4)-glucans having degrees of polymerization 9-31 and their quantitative analysis by high-performance anion exchange chromatography with pulsed amperometric detection. AB - Cyclic alpha-(1-->4)-glucans with degrees of polymerization (DPs) 9-31 were isolated from a mixture of cyclization products formed in the early stage of the action of cyclodextrin glucanotransferase (CGTase) on synthetic amylose, and characterized by matrix-assisted laser desorption ionization time-of-flight MS, 13C-NMR and HPLC of their partial acid hydrolyzates. High-performance anion exchange chromatography (HPAEC) with pulsed amperometric detection for an accurate estimation of cyclic alpha-(1-->4)-glucans was developed using those isolate glucans as quantification standards, and by HPAEC, the time course of the cyclization reaction of CGTase from an alkalophilic Bacillus sp. A2-5a on synthetic amylose was determined. PMID- 10481980 TI - Non-volatile products of triolein produced at frying temperatures characterized using liquid chromatography with online mass spectrometric detection. AB - Oxidation products from triolein under model heated frying conditions have been analyzed using liquid chromatography with an evaporative light scattering detector and atmospheric pressure chemical ionization (APCI) mass spectrometric detection. Triolein was heated at 190 degrees C with 2% water added each hour, to simulate the moisture of a frozen product, until polar components reached approximately 30%. The samples were separated using reversed-phase high performance liquid chromatography with APCI-MS detection. Triolein oxidation products included hydroperoxides, epoxides and a ketone. Other products were formed by shortening of an acyl chain on the intact triolein. Normal and oxygen containing products formed by the dimerization of triolein were also observed. Other products included chain addition products formed by addition of acyl chain subunits to intact triolein to form higher molecular weight products. PMID- 10481981 TI - High-performance liquid chromatographic methods for monitoring of isomers of 17 hydroxy-16-hydroxymethyl-3-methoxyestra-1,3,5(10)-triene. AB - Claisen condensation and consecutive reduction of 3-methoxyestra-1,3,5(10)-trien 17-one theoretically leads to four diastereomers of 17-hydroxy-16-hydroxymethyl-3 methoxyestra-1,3,5(10)-triene and their further transformations give different compounds with different biological activities. High-performance liquid chromatographic methods were developed for separation of the four isomers of 17 hydroxy-16-hydroxymethyl-3-methoxyestra-1,3,5(10)-triene: reversed-phase separation on a Nucleosil ODS C18 column with water-methanol as mobile phase; and normal-phase separation on an APEX Silica column with hexane-dichloromethane-2 propanol as mobile phase. The effects of eluent composition and flow-rate on the separation were investigated. This is the first chromatographic evidence for the formation of the 16alpha,17alpha isomer in the reduction of 16-hydroxymethylene-3 methoxyestra-1,3,5(10)-trien-17-one. PMID- 10481982 TI - High-yield purification of biosynthetic human growth hormone secreted in Escherichia coli periplasmic space. AB - A six-step, high-yield purification procedure for the preparation of clinical grade recombinant human growth hormone (rhGH) secreted in bacterial periplasmic space is described. Particular emphasis is given to hormone recovery yields and maximum contaminant host cell elimination. The strategy adopted, in addition to using one precipitation and five chromatographic steps in a particularly efficient sequence, was also based on running E. coli proteins - immunoradiometric assay profiles right after each chromatographic elution. Thus, an overall rhGH recovery higher than 40%, with a final concentration of E. coli proteins below 10 ppm is described for the first time. The accuracy of hGH and total protein quantification, especially in the early steps of the process, and the maximum elimination of hGH-related forms were also studied in detail. For these purposes size-exclusion and reversed-phase HPLC were found to be extremely valuable analytical tools. PMID- 10481983 TI - Optimization study for the reversed-phase ion-pair liquid chromatographic determination of nicotine in commercial tobacco products. AB - The availability of published methods for the determination of nicotine in commercial tobacco products based on state-of-the-art chromatographic methods is limited. Nicotine is a diprotic base with pKa's of 3.12 (pyridine ring) and 8.02 (pyrrolidine ring). Other monoprotic and diprotic bases are also present in commercial tobacco including anatabine, nornicotine, anabasine, and cotinine. In this paper, the chromatography of nicotine and the minor tobacco alkaloids under reversed-phase ion-pairing conditions is thoroughly studied. The results of this study are used to understand the retention mechanisms of the tobacco alkaloids, to examine their observed elution order with respect to fundamental analyte properties (size, functionality, and acid-base strength), and to select optimum chromatographic conditions for the determination of nicotine in commercial tobacco products. PMID- 10481984 TI - Quantification of phenylurea herbicides and their free and humic acid-associated metabolites in natural waters. AB - There is increasing interest in and demand for simultaneously monitoring pesticides as well as related degradation products (DPs) in natural waters, as the latter compounds can be even more toxic than the former ones. A method for determining parts per trillion levels of phenylurea herbicides and their DPs, that is their dealkylated forms and aromatic amines, is described. This method is based on solid-phase extraction with a Carbograph 4 cartridge followed by liquid chromatography (LC) with electrospray (ES) mass spectrometric detection. A study aimed at optimizing the response of the ES-MS detector for very weakly basic chloroanilines was conducted. Results showed that ion signal intensities of the above species were dependent on the composition of the LC mobile phase to an astonishing degree. At concentration levels of a few hundred ng/l, laboratory experiments showed that the aromatic amines considered here were mostly associated to dissolved humic acids (HAs) by both reversible and irreversible bindings. The addition of a reducing agent, i.e., NaBH4, succeeded in liberating that fraction of aromatic amines, which being reversibly bound to quinoidal structures of HAs are bioavailable. Analyte recoveries were better than 85% on extraction from 4 l of drinking water (spike level, 25 ng/l), 2 l of ground water (spike level, 50 ng/l) and 0.5 l of river water (spike level, 200 ng/l). Relative standard deviations ranged between 4.6 and 20% for drinking water, 4.3 and 15% for ground water, 5.9 and 13% for river water. Method detection limits calculated for drinking water, groundwater and surface water were between 3 and 11, 6 and 21, 36 and 75 ng/l, respectively. PMID- 10481985 TI - Separation and characterization of octylphenol ethoxylate surfactants used by reversed-phase high-performance liquid chromatography on branched fluorinated silica gel columns. AB - The separation and characterization of octylphenol ethoxylate surfactants were carried out by reversed-phase high-performance liquid chromatography on branched fluorinated silica gel columns. For Triton X-100, simultaneous separation of octylphenol ethoxylate oligomers, positional isomers of octylphenyl group and butylphenol ethoxylate oligomers was achieved. These oligomers were completely separated and identified by means of MS spectra. Ethoxylated oligomers are eluted in the sequence from small to large oligomers. Fifty-five oligomers of Triton X 405 could be separated by using gradient elution. To separate octylphenol ethoxylate surfactant, non-end-capped branched fluorinated silica gel columns were superior to end-capped columns. The relationship between ln k' and methanol concentration was linear, indicating that branched fluorinated silica gel columns were operating in the reversed-phase mode. As Van 't Hoff plots of capacity factor for all oligomers gave straight lines, the equilibrium of conformation for the ethylene oxide chain might lay to one side of either zigzag or meander conformers. PMID- 10481986 TI - Analysis of anions in aqueous samples by ion chromatography and capillary electrophoresis. A comparative study of peak modeling and validation criteria. AB - The object of this study is the comparison of two methods for the quantitative analysis of anions in aqueous samples: ion chromatography with conductimetric detection, and capillary zone electrophoresis with indirect photometric detection. The comparison includes modeling of experimental peaks as well as statistical validation criteria according to the recommendations of the International Conference on Harmonisation. In ion chromatography, peak shapes are Gaussian or exponentially modified Gaussian, and the number of theoretical plates calculated using the appropriate mathematical relations correspond well to those obtained from statistical moments. Peaks in capillary electrophoresis, however, do not follow the same models. A different model, treating the peaks as right angle triangles, has been studied. Equations corresponding to this model permit a good estimation of plate numbers. The statistical validation of these methods includes detection limits, linearity, accuracy and precision. Overall, ion chromatography yields better validation results than capillary electrophoresis. In the latter method the injection mode plays an important role, with voltage injection giving lower detection limits than hydrodynamic injection. PMID- 10481987 TI - Qualitative and quantitative analysis of new alkyl amide arginine surfactants by high-performance liquid chromatography and capillary electrophoresis. AB - A protocol for the qualitative and quantitative analysis of novel arginine-based cationic surfactants using HPLC and CE was studied and compared. The optimization of the analytical conditions was carried out through a systematic variation of the experimental parameters such as mobile phase, eluent conditions, ion pairing and amount of sample for HPLC, and type of buffer, ion strength, type and amount of organic solvent, sample injection time, applied voltage and column washing and conditioning for CE. PMID- 10481988 TI - Gas chromatographic kinetic study of carbon monoxide oxidation over platinum rhodium alloy catalysts. AB - The kinetics for the oxidation of carbon monoxide in the presence of excess oxygen over Pt-Rh alloy catalysts were studied by using the reversed-flow gas chromatography technique. Suitable mathematical analysis equations were derived by means of which the rate constants for the oxidation reaction of carbon monoxide, as well as for the adsorption and desorption of the reactant CO on the catalysts pure Pt, 75 atom% Pt+25 atom% Rh, 50 atom% Pt+50 atom% Rh, 25 atom% Pt+75 atom% Rh and pure Rh supported on SiO2 were determined. All the catalysts show a maximum rate constant for the production of CO2 at a characteristic temperature close to that found in the literature. The rate constants for the adsorption of CO increase generally with increasing temperature, while those for the desorption decrease with increasing temperature. From the variation of the rate constants with temperature activation energies for the oxidation reaction and adsorption of CO were determined, which are sensitive to the composition of the catalytic surface. The appearance of CO2 and carbon, when introducing pure CO into the column with the catalysts, verified a partial dissociative adsorption (e.g., disproportionation) of CO on the catalysts used. The latter indicates a mechanism for the CO oxidation through a partial dissociative adsorption of CO followed by the reaction of adsorbed CO molecules with adsorbed O atoms. PMID- 10481989 TI - Alkyl nitrates as achiral and chiral solute probes in gas chromatography. Novel properties of a beta-cyclodextrin derivative and characterization of its enantioselective forces. AB - Achiral and chiral interactions of alkyl nitrates (R-O-NO2) with heptakis-(3-O acetyl-2,6-di-O-pentyl)-beta-cyclodextrin (LIPODEX-D) in the gas phase were investigated chromatographically. Two major outcomes can be summarized. First, LIPODEX-D shows very fast temperature-dependent variations of the selectivity up to changes in the order of elution for C1-C5 alkyl nitrates. These changes in selectivity reveal that LIPODEX-D possesses different shape selectivities for small alkyl nitrates at different temperatures (40-80 degrees C), i.e. with increasing isothermal separation temperature extended (chain-like) alkyl nitrates have increased retention relative to bulky alkyl nitrates. The observations are highly reproducible and might indicate conformational changes of the cyclodextrin, however, an ultimate proof would require further spectroscopic investigation. Secondly, the chiral separations of systematically varied sets of C4-C11 alkyl nitrates allowed the thermodynamic characterization of enantiodiscriminating interactions. Quantitative evidence is provided showing that the presence of an ethyl group at the asymmetric carbon atom of an n-alkyl nitrate gives a strong enthalpic contribution to the resulting enantioselectivity. The Gibbs free energy differences -deltaR,S(deltaG) decrease systematically three to six times if the ethyl group is either shortened or enlarged by only a -CH2-increment. The results are based on two separate thermodynamic approaches, i.e. the determination of thermodynamic quantities ( deltaR,S(deltaG), -deltaR,S(deltaH), -deltaR,S(deltaS), Tiso) and a theoretical concept of enthalpy-entropy compensation. The data from our laboratory experiments also indicate that van der Waals interactions are responsible for chiral discrimination. PMID- 10481990 TI - Enhancement of electron-capture detection of methyl bromide in air by iodination. AB - An instrumentally simple and cost-effective method for the direct analysis of methyl bromide in ambient air is described. The method is based on the separation of sample components by gas chromatography, the conversion of methyl bromide to methyl iodide by reaction with an inorganic iodide salt, and the detection of the methyl iodide thereby produced by an electron-capture detector. Of the 20 different inorganic salts investigated here for conversion of methyl bromide to methyl iodide, zinc iodide was found to provide the greatest conversion efficiency. In addition, zinc iodide was found to provide high conversion efficiency at a modest reaction temperature, thereby minimizing both the thermal decomposition of compounds within the reaction volume and the level of column bleed introduced to the detector. The reactions of several other brominated and chlorinated organic compounds with zinc iodide have also been characterized. The successful application of this instrument to the quantitative determination of methyl bromide in a local background air sample is then demonstrated. PMID- 10481991 TI - Quantitative determination of polycyclic aromatic hydrocarbons, polychlorinated biphenyls and organochlorine pesticides in sewage sludges using supercritical fluid extraction and mass spectrometric detection. AB - Supercritical CO2 extraction (SFE) proved to be a valuable, fast, quantitative and partly selective extraction technique for determining polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and chlorinated pesticides (OCPs) in sewage sludges of different catchment areas based on a comparison study with certified reference materials (CRMs) and on recovery data. Most PCB extracts could be measured directly after SFE whereas for PAHs a short clean-up improved the final separation by high-resolution capillary gas chromatography. Due to the amount of coextracted compounds and the low levels of OCPs, extracts obtained by SFE had to be submitted to a multistep clean-up for final measurement. Average concentrations of 6.9 mg/kg dry mass (dm) for the sum of the 16 Environmental Protection Agency PAHs, 0.1 mg/kg dm for the sum of the seven PCB congeners and 0.002-0.072 mg/kg dm for the OCPs were found in the sewage sludges. Compared to studies performed earlier in our laboratory PAHs are still present in similar concentrations whereas PCB levels have decreased significantly. OCPs could be detected in only low amounts. A correlation between sludge type and degree of contamination could only be found for PAHs. PMID- 10481992 TI - pH effects on micelle-water partitioning determined by micellar electrokinetic chromatography. AB - In many biological and environmental situations, the pH of aqueous media varies and differences in solute partitioning may result. However, the majority of biopartitioning and hydrophobicity studies conducted have been at pH 7. Using migration factors measured by micellar electrokinetic chromatography, we have determined pH effects on micelle-water partitioning for 19 compounds. We develop an improvement to the migration factor equation and the corrected migration factor for aniline shows a definite increase as pH decreases. The corrected migration factor was constant for the rest of the compounds over the pH ranged studied. We also investigated five micelle markers and determined that decanophenone is the best micelle marker to date. Decanophenone has a strong chromophore, detectable at all pH levels, and is easy to dissolve in the mobile phase. PMID- 10481993 TI - Molecular weight determination of hyaluronic acid by gel filtration chromatography coupled to matrix-assisted laser desorption ionization mass spectrometry. AB - An analytical approach has been described for the molecular weight characterization of enzymatically degraded hyaluronic acid (HA). The approach involved the combined use of aqueous gel filtration chromatography (GFC) with matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). Microfractions were collected across an eluting peak from the chromatography system, followed by mass spectrometric analysis of these narrow fractions. The molecular mass determined by MALDI-MS and the signal obtained from the chromatography established a calibration curve for other hyaluronic acid samples analyzed by this GFC system. Results of one HA sample were obtained from both the calibration curve and direct fraction-by-fraction MALDI-MS analysis, and comparison of these results showed reasonable agreement. In contrast, molecular weights resulted from external calibration using dextran and pullullan standards showed drastically different numbers. Therefore, the GFC-MALDI-MS approach is a reliable method for the molecular weight characterization of polydisperse polysaccharides for which suitable calibration standards are unavailable for conventional GFC analysis. PMID- 10481994 TI - Revision of a theoretical expression for gas-liquid chromatographic retention. AB - In this communication, we revise some aspects of the [ideal gas/Van der Waals fluid] partition, derived in an earlier publication. The general character of the conclusions concerning the dependence of the partial molar free energy of solution, deltaGs(n), on the chain length, n, of linear solute molecules is shown through the relationship with more general partition formulations. Simultaneously, the correction of an error in the expression of the retention time dependence on the phase ratio of the chromatographic column, beta, is carried out. The misleading source of this error was redundant accounting in the solute translational contribution to deltaGs(n). PMID- 10481995 TI - Investigation of membrane dryers and evaluation of a new ozone scrubbing material for the sampling of organosulphur compounds in air. AB - The applicability of two different types of Nafion membrane dryers (based on counter-current flow and desiccant drying) and of a new ozone scrubbing material, polyphenylene sulphide wool (noXon-S), to adsorptive sampling of selected volatile sulphur compounds (methanethiol, dimethyl sulphide, isopropanethiol and isobutanethiol) is investigated at the low ppb (v/v) level (1-5 ppb). No analyte losses occur with either type of dryer at relative humidities (RH) of < or = 50%, while at higher RH values particularly the thiols tend to be lost (between 6 and 32%) even after conditioning. The actual losses depend more on the state of the individual permeation membrane rather than on the type of dryer. NoXon-S is a highly suitable ozone scrubber material for sulphur compounds since it efficiently removes ozone without retainment of the analytes and without the formation of blanks or artefacts from the scrubber material. The combined use of a Nafion membrane dryer and a noXon-S ozone scrubber is thus recommended for artifact-free sampling of sulphur compounds. PMID- 10481996 TI - Chemiluminescence detection of heme proteins separated by capillary isoelectric focusing. AB - Chemiluminescence detection was combined with capillary isoelectric focusing to perform protein analysis with high sensitivity. Luminol-H2O2 chemiluminescence was utilized, and heme proteins such as cytochrome c, myoglobin and peroxidase were analyzed. The proteins were focused by use of Pharmalyte 3-10 as ampholytes. Hydroxypropylmethyl-cellulose was added to the sample solution in order to easily reduce protein interactions with the capillary wall as well as the electroendoosmotic flow. The focused proteins were transported by salt mobilization to chemiluminescence detection cell equipped with an optical fiber. The present method showed significantly high sensitivity and wide dynamic range; the detection limit for cytochrome c was 6 x 10(-9) M and the linear dynamic range was greater than two-orders of magnitude (up to 2 x 10(-6) M). PMID- 10481997 TI - On the need to retain unregulated choice of high-performance liquid chromatography columns. PMID- 10481998 TI - Role of an A-type K+ conductance in the back-propagation of action potentials in the dendrites of hippocampal pyramidal neurons. AB - Action potentials elicited in the axon actively back-propagate into the dendritic tree. During this process their amplitudes can be modulated by internal and external factors. We used a compartmental model of a hippocampal CA1 pyramidal neuron to illustrate how this modulation could depend on (1) the properties of an A-type K+ conductance that is expressed at high density in hippocampal dendrites and (2) the relative timing of synaptic activation. The simulations suggest that the time relationship between pre- and postsynaptic activity could help regulate the amplitude of back-propagating action potentials, especially in the distal portion of the dendritic tree. PMID- 10481999 TI - Low-dimensional dynamics in sensory biology 2: facial cold receptors of the rat. AB - We report the results of a search for evidence of unstable periodic orbits in the sensory afferents of the facial cold receptors of the rat. Cold receptors are unique in that they exhibit a diversity of action potential firing patterns as well as pronounced transients in firing rate following rapid temperature changes. These characteristics are the result of an internal oscillator operating at the level of the membrane potential. If such oscillators have three or more degree of freedom, and at least one of which also exhibits a nonlinearity, they are potentially capable of complex activity. By detecting the existence of unstable periodic orbits, we demonstrate low-dimensional dynamical behavior whose characteristics depend on the temperature range, impulse pattern, and temperature transients. PMID- 10482000 TI - Silicon synaptic conductances. AB - We have developed compact analog integrated circuits that simulate two synaptic excitatory conductances. A four-transistor circuit captures the dynamics of an excitatory postsynaptic current caused by a real AMPA conductance. A six transistor circuit simulates the effects of a real voltage-dependent NMDA conductance. The postsynaptic current dynamics are modeled by a current mirror integrator with adjustable gain. The voltage dependence of the silicon NMDA conductance is realized by a differential pair. We show the operation of these silicon synaptic conductances and their integration with the silicon neuron (Mahowald and Douglas, 1991). PMID- 10482001 TI - Role of calcium electrogenesis in apical dendrites: generation of intrinsic oscillations by an axial current. AB - Dendrites are covered with conductances whose function is still mysterious. Using intracellular recording and calcium imaging, we describe an electrogenic band of calcium channels in distal apical dendrites of layer 5 pyramidal neurons (Yuste et al., 1994). We now explore the functional consequences of this distal electrogenic area with multicompartmental numerical simulations. A calcium imaging and electrophysiological database from a single neuron, recorded under blocked sodium and potassium conductances, is replicated by simulations having increased dendritic calcium current. In these models a significant axial current flows from the apical dendrite into the somatic region, activating low-threshold calcium channels and generating oscillations similar to those seen in the electrophysiological data. We propose that the distal electrogenic area in apical dendrites serves to inject current into the soma and produce intrinsic oscillatory dynamics. PMID- 10482002 TI - Analysis and modeling of frequency-specific habituation of the goldfish vestibulo ocular reflex. AB - Modification of the vestibulo-ocular reflex (VOR) by vestibular habituation is an important paradigm in the study of neural plasticity. The VOR is responsible for rotating the eyes to maintain the direction of gaze during head rotation. The response of the VOR to sinusoidal rotation is quantified by its gain (eye rotational velocity/head rotational velocity) and phase difference (eye velocity phase--inverted head velocity phase). The frequency response of the VOR in naive animals has been previously modeled as a high-pass filter (HPF). A HPF passes signals above its corner frequency with gain 1 and phase 0 but decreases gain and increases phase lead (positive phase difference) as signal frequency decreases below its corner frequency. Modification of the VOR by habituation occurs after prolonged low-frequency rotation in the dark. Habituation causes a reduction in low-frequency VOR gain and has been simulated by increasing the corner frequency of the HPF model. This decreases gain not only at the habituating frequency but further decreases gain at all frequencies below the new corner frequency. It also causes phase lead to increase at all frequencies below the new corner frequency (up to some asymptotic value). We show that habituation of the goldfish VOR is not a broad frequency phenomena but is frequency specific. A decrease in VOR gain is produced primarily at the habituating frequency, and there is an increase in phase lead at nearby higher frequencies and a decrease in phase lead at nearby lower frequencies (phase crossover). Both the phase crossover and the frequency specific gain decrease make it impossible to simulate habituation of the VOR simply by increasing the corner frequency of the HPF model. The simplest way to simulate our data is to subtract the output of a band-pass filter (BPF) from the output of the HPF model of the naive VOR. A BPF passes signals over a limited frequency range only. A BPF decreases gain and imparts a phase lag and lead, respectively, as frequency increases and decreases outside this range. Our model produces both the specific decrease in gain at the habituating frequency, and the phase crossover centered on the frequency of habituation. Our results suggest that VOR habituation may be similar to VOR adaptation (in which VOR modification is produced by visual-vestibular mismatch) in that both are frequency-specific phenomena. PMID- 10482003 TI - Temporal dispersion windows in cortical neurons. AB - A temporal dispersion window is the time required for a volley of action potentials on presynaptic axons to cross the dendritic arbor of a postsynaptic neuron. The volley produces a series of unitary postsynaptic potentials (PSPs) on the postsynaptic neuron. Temporal dispersion is, thus, one factor that can influence the integration of unitary PSPs and the production of action potentials in cortical neurons. Temporal dispersion windows for neurons in the visual cortex of the freshwater turtle, Pseudemys scripta, were estimated by characterizing geniculate afferents and the morphology of neurons in the visual cortex. Horseradish peroxidase injections in the thalamus revealed thin and unmyelinated terminal arbors that run horizontally from lateral to medial across the cortex, forming en passant synapses across the dendrites of cortical neurons. Axons with two calibers were seen, one with diameters between 0.5 and 2.0 microm, and a second with diameters below the resolution limit of the light microscope. The conduction velocity of geniculate afferents in the cortex was measured at 0.18 m/sec +/-0.04 using the latency of extracellular field potentials evoked by electrical stimulation of the lateral forebrain bundle. The positions and dendritic arbors were characterized in Golgi preparations. Seven morphologically distinct neuron types were positioned to intersect the geniculate afferents in Golgi preparations. The spatial overlap between the dendritic arbors of these cells and the geniculate afferents varied from 128 to 850 microm. Temporal dispersion windows for the seven cell types ranged from 0.7 to 4.7 msec, estimated using a geniculate fiber conduction velocity of 0.18 m/sec. Estimated conduction velocities of 0.04 m/sec for small-caliber fibers produce temporal dispersion windows of 3.2 to 21.3 m/sec. PMID- 10482004 TI - The SCID-hu xenogeneic transplantation model: complex but telling. AB - Animal models based on knock-out or transgenic technology are widely used in basic and applied biomedical research. An alternative to these approaches is the generation of xenogeneic transplantation models allowing the in-vivo investigation of cell types and organs. In the field of dermatology transplantation of human skin onto mice lacking functional B and T cells (SCID mice) and subsequent manipulation of these grafts yielded new insights in many different aspects of skin biology. This review highlights some of the applications of this versatile model focussing on phenomena relevant for the subject of dermatology. PMID- 10482005 TI - Presence of circulating antibodies to a disease-specific antigen on cultured human dermal microvascular endothelial cells in patients with Behcet's disease. AB - Behcet's disease is a chronic, multisystem disorder characterized by a recurrent inflammatory reaction. Antiendothelial cell antibodies have been detected in the serum from patients with autoimmune diseases with presenting vasculitis and it is assumed that they can induce damage to the endothelial cells. In this study, we detected antiendothelial cell antibodies in the serum from patients with Behcet's disease using an enzyme-linked immunosorbent assay, electrophoresis and immunoblotting. The cytolysis of human dermal microvascular endothelial cells (HDMEC) was measured using a cytotoxicity assay. The serum from 37.4% of Behcet's disease patients showed IgM antibodies against unstimulated HDMEC while the serum from 18.4% of patients showed an increase in IgM antibody titer after IFN-gamma pretreatment. The frequency of vasculitis was higher in the IgM-positive Behcet's disease patients than in the IgM-negative patients. In Western blotting, IgM positive Behcet's disease serum reacted with the 44 kDa HDMEC surface antigen, whereas IgM-positive systemic lupus erythematosus serum reacted with the 81 kDa HDMEC surface antigen. The reactivity to the 44 kDa protein band was also observed in cultured human umbilical vein endothelial cells but not in fibroblasts, A431 cells or SK-MEL-2 cells. Serum from Behcet's disease patients incubated with human complement or mononuclear cells produced no significant lysis of HDMEC, and cultured HDMEC were resistant to antibody-dependent cell mediated cytotoxicity. The results suggest that antibodies against antigens on the surface of endothelial cells may play a role in inducing vasculitis in Behcet's disease, not through a direct toxic effect of an antiendothelial cell antibody but by an indirect effect involving the activation of endothelial cells to produce cytokines. PMID- 10482007 TI - Clobetasol-17 propionate lotion under hydrocolloid dressing (Duoderm ET) once weekly versus unoccluded clobetasol-17-propionate ointment twice daily in psoriasis: an immunohistochemical study on remission and relapse. AB - It is well established that the efficacy of corticosteroids under occlusion with hydrocolloids (HCD) is superior compared to monotherapy with topical corticosteroids. However, following treatment with more potent corticosteroids, increased side effects and a more pronounced rebound might be expected. In the present clinical study, the efficacy of relapse after and the safety characteristics of two treatment modalities were compared: clobetasol-17 propionate lotion under an HCD dressing once weekly versus clobetasol-17 propionate ointment without an HCD twice daily. Clinical assessments were recorded and skin biopsies were taken before therapy, at clearance and 6 weeks after clearance. A panel of monoclonal antibodies to characterize epidermal proliferation, differentiation and inflammation were selected. In addition, clinical and histological assessments for skin atrophy were made. Both therapies had a major therapeutic effect, which was reflected in the clinical and immunohistochemical parameters. The only difference between the two therapies was a faster remission induction time in patients treated with corticosteroids combined with HCD. Six weeks after discontinuation of treatment, similar clinical and histological signs of relapse were observed for both therapies. Clinically, there were no signs of skin atrophy but histologically, epidermal thinning occurred to the same extent with both therapies but proved to be reversible within 6 weeks of discontinuation of treatment. From this study it can be concluded that the combination of HCD and corticosteroids is able to induce relatively fast remission compared to corticosteroid monotherapy but relapse and safety characteristics are comparable to the unoccluded corticosteroid therapy. PMID- 10482006 TI - Hyporesponsiveness of peripheral blood lymphocytes to streptococcal superantigens in patients with guttate psoriasis: evidence for systemic stimulation of T cells with superantigens released from focally infecting Streptococcus pyogenes. AB - Throat infection with Streptococcus pyogenes is the most important trigger for acute guttate psoriasis. We examined the in vitro responses of peripheral blood mononuclear cells (PBMC) to streptococcal superantigens, SPEA and SPEC, and staphylococcal superantigens, SEB and TSST-1, in patients with guttate psoriasis, in patients with chronic plaque psoriasis, and in healthy subjects. PBMC from patients with guttate psoriasis responded poorly to SPEA and SPEC at concentrations of 0.1 and 1 ng/ml as compared with those from patients with plaque psoriasis, but showed high responses to SEB and TSST-1. The hyporesponsiveness recovered after improvement of the skin eruption. There was no significant difference between guttate and chronic types of psoriasis in the percentage of circulating T-cell receptor BV2 or BV8-bearing T cells, responsive to streptococcal superantigens, indicating that T-cell clonal anergy was a mechanism underlying the hyporesponsiveness. Our results suggest that superantigens released from focally infecting S. pyogenes induce a transient activation of relevant T cells, leading to the development of skin eruption and, subsequently, temporary T-cell anergy to these toxins. PMID- 10482008 TI - The dermoepidermal junction in psoriatic skin as revealed by scanning electron microscopy. AB - Our previous ultrastructural and immunohistochemical studies, in vivo and in vitro, have shown important modifications of the basement membrane of psoriatic skin, which could play a key role in the alterations of keratinocyte adhesion, migration, proliferation and differentiation. In order to complete the morphological examination of all the structures in the dermoepidermal junction of psoriatic skin, we carried out a scanning electron microscopic study using biopsies taken from eight psoriatic patients. The biopsies were fixed in a mixture of 0.2% paraformaldehyde and 0.25% glutaraldehyde in 0.1 M cacodylate buffer at pH 7.4. The specimens were then frozen in liquid nitrogen and fractured following the natural cleavage planes and observed under a Jeol JSM-6301F field emission scanning electron microscope operating at 1.8-2.0 kV. The basal keratinocytes observed showed pore-like depressions on the lateral plasmalemma and villous-like projections in very dilated intercellular spaces. Moreover the basal cell plasma membrane was seen to rest on the papillary dermis without interposition of the lamina densa. The detachment of some keratinocytes enabled the examination of the lamina densa, which appeared slightly granular with numerous focal interruptions through which it was possible to observe the underlying collagen fibres. These findings, together with previously reported findings, support the hypothesis that in psoriasis molecular and structural alterations of the dermoepidermal junction are present, that could fundamentally alter the regulation of the cytomorphological processes and the normal functions of the basement membrane. PMID- 10482009 TI - In vivo introduction of the interleukin 6 gene into human keratinocytes: induction of epidermal proliferation by the fully spliced form of interleukin 6, but not by the alternatively spliced form. AB - The achievement of keratinocyte gene therapy in clinical practice requires fundamental experiments using human keratinocytes or skin. We have recently demonstrated that the in vivo introduction of the interleukin 6 (IL-6) gene into rat keratinocytes induces epidermal proliferation and lymphocyte infiltration into the skin. In this study, we first amplified the human IL-6 cDNA from oligo dT-primed keratinocyte cDNA and then detected the fully spliced (FS) form and the alternatively spliced (AS) form of IL-6 cDNA. Sequence analysis showed that the AS form, which was composed of the IL-6 coding region with all of exon II deleted except for the first guanine, was identical to that reported to be present in lymphocytes. We constructed the expression vectors phIL6 of the FS form and phIL6S of the AS form. We transplanted human skin onto nude rats and introduced phIL6 and phIL6S into the human keratinocytes using the naked DNA method. Keratinocytes prepared 24 h after introduction from the areas treated with them were examined by reverse transcriptase (RT)-PCR and enzyme linked immunosorbent assay (ELISA). RT-PCR showed that the amounts of FS IL-6 mRNA and AS IL-6 mRNA were similar, whereas the ELISA showed that the amount of FS IL-6 peptide was four times that of the AS IL-6 peptide. Histological examination 48 h after introduction showed that the FS form had induced epidermal proliferation, whereas the AS form had not. The epidermal thickening without lymphocyte infiltration induce by the FS form indicates that keratinocyte proliferation is caused by a direct effect of overexpressed IL-6, and not by a secondary effect of infiltrating lymphocytes. This is the first report of the introduction of a human gene into human keratinocytes to produce a biologically active transgenic gene product in human skin using the naked DNA method. PMID- 10482010 TI - Incorporation of linoleic acid by cultured human keratinocytes. AB - Linoleic acid is required for the formation and maintenance of the epidermal barrier, but most of the current in vitro keratinocyte culture systems are linoleic acid-deficient. The aim of the present study was to examine the efficiency of linoleic acid uptake in human keratinocyte cultures grown under submerged and air-exposed conditions in serum-free medium. The water-insoluble linoleic acid was bound to carrier molecules (cyclodextrin or bovine serum albumin). Comparable results were obtained with home-made and commercially available linoleic acid complexes. In the submerged cultures, the increase of the linoleic acid medium concentration (ranging from 0 to 20 microg/ml) resulted in a gradual increase in the linoleic acid cellular content, which exceeded 1.4 times the value found in native epidermis when the highest concentration of linoleic acid was used. The addition of linoleic acid did not alter the profile of the other epidermal fatty acids, with the exception of oleic acid, which decreased in parallel with the increasing linoleic acid content. While the content of linoleic acid found in phospholipids was similar to that in native epidermis, a large excess of linoleic acid was detected in triglycerides, the synthesis of which was markedly increased in cultures grown submerged in medium containing higher concentrations of linoleic acid. Under air-exposed conditions, the dermal substrate used seemed to be the most limiting factor for efficient linoleic acid supplementation. A low linoleic acid cellular content was detected when an inert filter was used. De-epidermized dermis was found to be the most permeable substrate for linoleic acid complexes. The cellular linoleic acid content increased in a parallel with the increasing linoleic acid concentration (ranging from 4 to 30 microg/ml), but the overall amount incorporated was lower than that in submerged cultures. The content of linoleic acid in the phospholipid and ceramide fractions isolated from reconstructed epidermis grown under air-exposed conditions was close to that of native epidermis, but the triglycerides remained abnormally enriched in linoleic acid, indicating persistence of some anomalies in epidermal lipogenesis in vitro. PMID- 10482011 TI - Immunohistochemical staining of proliferating cell nuclear antigen (PCNA) in malignant and nonmalignant skin diseases. AB - Immunohistochemical staining of proliferating cell nuclear antigen (PCNA) was performed in skin from patients with various malignant and nonmalignant skin diseases using anti-PCNA monoclonal antibodies. The malignant diseases included squamous cell carcinoma (SCC), adult T lymphotrophic leukemia (ATL), mycosis fungoides, malignant melanoma and malignant lymphoma, and the nonmalignant diseases included severe treatment-resistant atopic dermatitis (AD), psoriasis vulgaris, verruca vulgaris, and others. The percentage of PCNA-positive cells (the labeling index, LI) was highest for the malignant diseases (56.5+/-7.1%). The LIs for severe treatment-resistant AD, psoriasis, and verruca vulgaris were also significantly higher than those for the normal control or nonlesional skin of the patients. The PCNA LIs were, however, not significantly elevated in eczema and contact dermatitis. The high PCNA LIs in severe AD and psoriasis vulgaris were considerably lower in the skin improved by treatment. Labeling with Ki67, a nuclear protein expressed in cycling cells, was also performed in skin from subsets of each patient group. The results were very similar to those found with PCNA labeling. PCNA-positive cells were found throughout the dermis as well as the basal layer in the malignant diseases, whereas they were found only in the basal layer in the nonmalignant diseases. The results suggest that in human skin diseases, the extent of staining for PCNA, which is a cofactor of DNA polymerase delta and is essential for cell proliferation, correlates with the extent to which the disease is treatment-resistant. In addition, our findings suggest that the PCNA LI and distribution of PCNA-positive cells in the skin may be helpful in the early diagnosis of skin malignancies. PMID- 10482012 TI - Neuronal structural proteins, transmitters, transmitter enzymes and neuropeptides in human Meissner's corpuscles: a reappraisal using immunohistochemistry. AB - The innervation of Meissner's corpuscles (Mc) is complex, consisting of different types of sensory nerve fibers. We investigated the neurochemistry of Mc in human digital skin by indirect immunofluorescence, using a wide panel of both general neuronal as well as neurotransmitter-related molecules. Structural proteins (protein gene product 9.5, neuron-specific enolase, neurofilament) were found to consistently label the entire neuronal component of Mc. Immunoreactivity for gamma-melanocyte stimulating hormone was detected in the large diameter fibers running spirally within the corpuscles, while a number of peptide transmitters (substance P, calcitonin gene-related peptide, neurokinin A, galanin, somatostatin) were found in the thin unmyelinated fibers in both intra- and extracorpuscular locations. PMID- 10482013 TI - Airborne Compositae dermatitis: monoterpenes and no parthenolide are released from flowering Tanacetum parthenium (feverfew) plants. AB - The air around intact feverfew (Tanacetum parthenium) plants was examined for the presence of airborne parthenolide and other potential allergens using a high volume air sampler and a dynamic headspace technique. No particle-bound parthenolide was detected in the former. Among volatiles emitted from the aerial parts of feverfew plants and collected by the dynamic headspace technique a total of 41 compounds, mainly monoterpenes, were identified and quantified by GC and GC MS. Alpha-Pinene, camphene, limonene, gamma-terpinene, (E)-beta-ocimene, linalool, p-cymene, (E)-chrysanthenol, camphor and (E)-chrysanthenyl acetate were the predominant monoterpenes accounting for nearly 88% of the total volatiles emitted. The average total yield of volatiles emitted over 24 h was 18,160 ng/g fresh weight of leaves and flowers, corresponding to the emission of approximately 8 mg volatiles per day from one full-grown feverfew plant. No parthenolide or other sesquiterpene lactones were detected. The present investigation does not support the theory of airborne sesquiterpene lactone containing plant parts or of direct release of sesquiterpene lactones from living plants as the only explanations for airborne Compositae dermatitis. Potential allergens were found among the emitted monoterpenes and their importance in airborne Compositae dermatitis is discussed. PMID- 10482014 TI - Alginate oligosaccharides modulate cell morphology, cell proliferation and collagen expression in human skin fibroblasts in vitro. AB - Effects of alginate oligosaccharides on cell proliferation and expression of collagen in cultured skin fibroblasts were studied. The oligosaccharides were found to suppress fibroblast proliferation to half the level in control cultures at a dose of 10 mg/ml during a period of 5 days. The inhibition was accompanied by a change in cell shape. The inhibition of cell proliferation was reversible, since depletion of these oligosaccharides led to a recovery of cell motility. Treatment of confluent cells with 10 mg/ml oligosaccharides for 5 days resulted in a reduction in collagen synthesis to one half of that in control cultures and inhibition of steady state levels of alpha1(I), alpha2(I), alpha1(III) and alpha1(VI) collagen mRNAs. These results suggest that alginate oligosaccharides are potential modulators of dermal fibroblasts and may provide a useful tool for the treatment of disorders related to abnormal collagen metabolism. PMID- 10482015 TI - Diversity of desmosomal proteins in regenerating epidermis: immunohistochemical study using a human skin organ culture model. AB - We recently established a skin organ culture model for epithelial healing by creating a central defect in freshly excised human skin specimens and keeping them in culture for up to 7 days, either untreated or with transplantation of allogenic or autologous keratinocytes. In this study the molecular diversity of cell-cell junction proteins in the regenerating epidermis was analysed immunohistochemically using a broad spectrum of monoclonal antibodies against glycoproteins (cadherins) and plaque proteins of desmosomes. At all stages studied the entire set of desmosomal cadherins [desmogleins (Dsg) 1-3 and desmocollins (Dsc) 1-3] was detected, with Dsg3, Dsc2 and Dsc3 being the most prominent. In the disordered neoepithelium at day 3 (after transplantation) some desmosomal cadherins appeared in their respective stratum compartments. In regenerating epidermis on day 7, which exhibited a more ordered stratification and a compact horny layer, stratification-related patterns of desmosomal cadherins were more pronounced. However, some immaturity of the day-7 neoepidermis was reflected by relatively low levels of the maturation-associated Dsgl and Dsc1 and a strong basal layer expression of Dsg2 which is sparse in normal epidermis. Desmosomal plaque proteins showed expression patterns similar to those in normal healthy epidermis. The adherens junction-related E-cadherin was also detected. Dendritic cells (melanocytes, Langerhans cells) were mainly present at the wound margins. In conclusion, this study demonstrated partial but not complete epidermal maturation and junction development during regeneration up to day 7. This model should also be useful in future studies to evaluate the effects of growth hormones to be used in therapeutic trials on chronic leg ulcers. PMID- 10482016 TI - Inhibition of contact sensitizer-induced migration of human Langerhans cells by matrix metalloproteinase inhibitors. AB - Emigration of Langerhans cells (LC) from the epidermis upon exposure to contact sensitizers is regarded as an essential event in the development of contact sensitization. Since migration of several types of cells depends on the activity of matrix metalloproteinases (MMPs), in the present study we tested whether MMP inhibitors (BB94, BB2116 and CT1166) can prevent the emigration of LC in cultured skin explants, which were exposed to a contact sensitizer (NiSO4). Epicutaneous application of NiSO4 significantly reduced the number of LC within the epidermis and the remaining LC were localized along the epidermal-dermal junction indicating the emigration of LC. In the presence of each of the MMP inhibitors tested, NiSO4-induced migration of LC was strongly decreased. Since after the epicutaneous application of contact sensitizer, its presentation by skin LC is essential for the development of contact sensitization instead of the development of antigen-specific tolerance, our results suggest that the use of MMP inhibitors may be beneficial for the prevention of contact sensitization of the host. PMID- 10482017 TI - Induction by tumour necrosis factor alpha of dose-related changes in Langerhans cell frequency in mice. AB - It has been demonstrated previously that tumour necrosis factor alpha (TNF-alpha) provides an important signal for the migration of epidermal Langerhans cells (LC) from the skin. Intradermal administration to mice of homologous recombinant TNF alpha induces both a rapid reduction in the frequency of LC local to the site of exposure and, somewhat later, an accumulation of dendritic cells in draining lymph nodes. It has been proposed recently, however, that the influence of TNF alpha on LC function may be dose-dependent in nature with lower concentrations inducing migration, but higher concentrations immobilizing LC in the epidermis. To investigate this proposal we examined the kinetics and dose-response relationships of TNF-alpha-induced LC migration in mice. At all concentrations tested (50, 150 or 300 ng/ear), intradermal exposure to TNF-alpha caused within 30 min a significant reduction in the frequency of MHC class II (Ia)+ LC within epidermal sheets. With the lower concentrations of TNF-alpha this effect was still apparent when LC were enumerated in the epidermis up to 4 h following cytokine treatment. In contrast, however, exposure of mice to 300 ng of TNF-alpha was consistently associated with a considerably less marked, and statistically insignificant, reduction in LC frequency by 4 h. These data indicate that at all concentrations of the cytokine examined here, TNF-alpha was able to stimulate a rapid (within 30 min) reduction in epidermal LC numbers, but that the rapidity with which the epidermis was repopulated following the initiation of LC migration was influenced by the concentration of TNF-alpha administered. It is suggested that TNF-alpha may influence not only the tempo of LC migration, but also the kinetics of epidermal repopulation. PMID- 10482018 TI - The effect of UV-B irradiation on primary and secondary HSV-1 infections in interleukin-4 knockout mice. AB - Ultraviolet B (UV-B) irradiation suppresses cell-mediated immunity and may lead to increased susceptibility to infectious diseases. Limited evidence suggests that exposure promotes a T helper (Th) 2 type of cytokine response with abrogation of a Th1 response. Several putative mediators of UV-induced immunosuppression have been identified, of which interleukin-4 (IL-4), an example of a Th2 cytokine, is one. Primary and secondary epidermal infections with herpes simplex virus (HSV) type 1 in IL-4 knockout (IL-4-/-) mice and the parent strain Bb 129 strain (IL-4+/+) were investigated using clinical features, phenotyping of cells from lymph nodes draining the sites of infection and lymphoproliferation assays. The IL-4-/- mice were more susceptible to both primary and secondary HSV infections than the parent mice. The percentage of lymph node dendritic cells (DC) expressing Ia was 45 in the IL-4+/+ mice but only 18 in the IL-4-/- strain, and the lymph node cells from infected IL-4-/- mice were less able to respond in vitro to HSV than those from the parent strain. Following suberythemal UV-B irradiation, more severe primary and secondary lesions resulted in both strains. There were fewer lymph node DC expressing Ia in both strains and this change was accompanied by suppression of the lymphoproliferation induced by HSV which was due to an effect on DC function rather than on the proliferative ability of the responding lymphocytes. UV-B exposure had no effect on ICAM-1 or B7.2 expression on the DC. Thus IL-4 seems to protect mice against HSV infection, and no evidence was obtained for the involvement of IL-4 in the UV-induced immunomodulation which results in more severe cutaneous HSV infections. PMID- 10482019 TI - Substance P- and antigen-induced release of leukotriene B4, prostaglandin D2 and histamine from guinea pig skin by different mechanisms in vitro. AB - Substance P (SP) induces increased vascular permeability, vasodilatation and granulocyte infiltration upon intradermal injection. Studies with antagonists and mast cell-deficient mice have suggested that granulocyte infiltration in response SP is mediated by leukotriene (LT) B4 derived from mast cells. However, the release of LTB4 has not been detected using mast cells isolated from human skin. Here we report the release of LTB4, prostaglandin (PG) D2 and histamine from guinea pig skin tissue in response to SP. The release of these agents occurred in a dose-dependent manner over a concentration range of SP from 1 x 10(-6) to 3 x 10(-4) M. No detectable PGE2 was released at any concentration up to 3 x 10(-4) M SP. The kinetics of histamine release induced in response to SP was more rapid than that induced by antigen. By comparison, SP-induced and antigen-induced release of LTB4 and PGD2 were similar, but slower than the histamine release. In the absence of extracellular Ca2+, release of histamine and PGD2 in response to SP was partially impaired, but to a lesser extent than that induced by antigen. On the other hand, LTB4 release in response to both SP and antigen was abolished under the same conditions. These results indicate that SP induces the release of LTB4, as well as histamine and PGD2, in the skin most likely from mast cells by a mechanism which may be different from that of mediator release in response to antigen. PMID- 10482020 TI - Analysis of the tumor suppressor gene p16(INK4A) in microdissected melanoma metastases by sequencing, and microsatellite and methylation screening. PMID- 10482021 TI - A Dutch approach to methicillin-resistant Staphylococcus aureus. PMID- 10482022 TI - Nephropathia epidemica and Puumala virus in Austria. AB - To study the epidemiology of hantavirus infections in Austria, 1215 humans and 596 rodents of different species were tested for the presence of antibodies against Puumala and Hantaan virus. Direct virus identification by polymerase chain reaction in lung tissue of serologically positive rodents was performed to verify antibody results and to determine the genetic identity of viral RNA by phylogenetic analysis of a part of the hantavirus M segment. For 32 of the 37 cases of nephropathia epidemica diagnosed in Austria, the location where transmission took place could be traced to specific areas in the Austrian federal states of Carinthia and Styria. The overall seroprevalence in humans was 1.2% and ranged from 0.02% in Villach, Carinthia, to 0.8% in Korneuburg, Lower Austria, and 1.8% in Wolfsberg, Carinthia. Virus RNA could be amplified from three Clethrionomys glareolus voles collected in Klippitztorl, Carinthia, and from one collected in Ernstbrunn, Lower Austria. The sequences were all identified as Puumala virus by phylogenetic analysis and were found to be most closely related to the western European Puumala viruses from Germany and France. No evidence of the existence of Hantaan-like infections and viruses in Austria was found. PMID- 10482023 TI - Pseudomonas aeruginosa bacteremia in patients infected with human immunodeficiency virus type 1. AB - A prospective analysis of 43 episodes of Pseudomonas aeruginosa bacteremia in HIV 1-infected subjects was performed and the results compared with the incidence and outcome of Pseudomonas aeruginosa bacteremia in other high-risk patients, such as transplant recipients, leukemia patients, or patients hospitalized in the intensive care unit. The incidence of bacteremia/fungemia as a whole and of gram negative and Pseudomonas aeruginosa bacteremia in particular was greater in HIV-1 infected subjects than in the unselected general population admitted. In contrast, the incidence of Pseudomonas aeruginosa bacteremia in HIV-1-infected patients did not differ from that in patients with other high-risk conditions. In patients with HIV-1 infection, independent risk factors for presenting Pseudomonas aeruginosa bacteremia were nosocomial origin (OR, 2.7; 95% CI, 1.3 5.7), neutropenia (OR, 2.7; 95% CI, 1.07-6.8), previous treatment with cephalosporins (OR, 3.6; 95% CI, 1.1-11.6), and a CD4+ cell count lower than 50 cells/mm3 (OR, 3.1; 95% CI, 1.7-8.6). Primary bacteremia and pneumonia were the most common forms of presentation. Fourteen (33%) patients died as a consequence of the bacteremia. The presence of severe sepsis (OR, 17.5; 95% CI, 3.2-68) and the institution of inappropriate definitive antibiotic therapy (OR, 2.7; 95% CI, 1.1-13) were independently associated with a poor outcome. One year after the development of bacteremia, only eight (19%) patients remained alive. PMID- 10482024 TI - Candidemia in HIV-infected subjects. AB - The epidemiological features of 37 episodes of candidemia in HIV-infected subjects were analysed in a retrospective matched case-control study conducted over an 8-year period (1990-1997). Univariate analysis identified eight risk factors that were significantly associated with candidemia (P<0.05): i) use of central venous catheters; ii) administration of total parenteral nutrition; iii) previous antifungal therapy; iv) previous therapy with glycopeptides; v) presence of oral/ esophageal candidiasis; vi) concomitant bacterial infections; vii) neutropenia; and viii) concomitant AIDS dementia complex. Stepwise logistic regression analysis revealed that the only independent risk factor for developing candidemia was the use of central venous catheters (P = 0.0001). Candida albicans was the most frequently isolated pathogen, accounting for 18 (48%) episodes of candidemia, followed by Candida tropicalis (19%) and Candida glabrata (11%). The crude mortality rate was 62%. On univariate analysis concomitant opportunistic infections, presence of non-Candida albicans species of Candida and neutropenia were shown to be predictive of death. Multivariate analysis revealed that the presence of non-Candida albicans strains of Candida was the only significant factor associated with a worse prognosis (P = 0.001). In conclusion, candidemia appears to be more common in patients with advanced HIV disease. Of the factors which influenced the onset of candidemia, use of central venous catheters seemed to be the most important one. PMID- 10482025 TI - Evidence-based diagnosis of Lyme disease. AB - The aim of this study was to make an evidence-based comparison of four commercial enzyme immunoassays (EIAs) (Serion Classics, Sigma Diagnostics, Cambridge Biotech and ICN Diagnostics) and an in-house enzyme immunoassay (EIA) in order to select the most appropriate screening assay for diagnosis of Lyme disease. Borrelia burgdorferi sensu stricto cultured in BSK-H medium was used to develop the in house assay. Escherichia coli antigen (0.9 mg/ml) was included in the serum diluent to reduce non-specific background. Comparison of the number of tests needed to diagnose (i.e. to indicate a positive result) and the cost per positive diagnosis for the five assays was made using a panel of 176 Western blot characterised sera. The Cambridge Biotech and Sigma assays had the highest sensitivity but poorer specificity, whereas the Serion and ICN assays had highest specificity but poorer sensitivity. The in-house assay had average sensitivity and specificity, the number of tests needed to diagnose being 2.32 compared to 1.92 for Serion, 2.17 for ICN, 2.5 for Sigma and 2.7 for Cambridge Biotech. In a diagnostic protocol that uses an EIA as screening test, with confirmation by Western blot, a good balance of sensitivity and specificity is essential. The in house assay was the most cost-effective (lowest cost per positive diagnosis), and is probably the best option for specialist laboratories in Europe. PMID- 10482026 TI - Growth characteristics and influence of antibiotics on rough/smooth phenotypic variants of Helicobacter pylori. AB - Helicobacter pylori shows a rather high variability of several biochemical markers including lipopolysaccharide structures. This study aimed to determine whether Helicobacter pylori has a potential for phenotypic variability and to describe its effects on bacterial pathogenesis. From colonies of three clinical strains of Helicobacter pylori with rough (R) colony morphology, spontaneous phenotypic variants with smooth (S) colony morphology were isolated that occurred with a frequency of 10(-2) to 10(-3), irrespective of growth conditions. R variant bacteria produced exclusively low-molecular-mass lipopolysaccharide. They exhibited increased lysis in the presence of plain air. In contrast, the S variants produced low- and high-molecular-mass lipopolysaccharide and did not exhibit increased lysis in the presence of plain air. Cocultivation of bacterial cells with AGS stomach cancer cells revealed that R-variant bacteria but not S variant bacteria effected an inhibition of high molecular-weight glycoprotein biosynthesis and secretion by the host cells. Skirrow supplement added as selective agent to liquid and/or solid media was tolerated to a similar extent among R- and S-variant bacteria, while all variants proved sensitive to metronidazole, amoxicillin and clarithromycin except for the R and S isolates of strain Hp57, which showed resistance to the latter compound. It was concluded that R- and S-variants of Helicobacter pylori may have distinct roles in pathogenesis; nevertheless, these bacteria may be isolated by traditional methods and eradicated by conventional anti-infective therapy. PMID- 10482027 TI - Campylobacter bacteremia and pneumonia in two splenectomized patients. AB - Two patients with beta thalassemia who had undergone splenectomy for hypersplenism were admitted to the hospital with high fever and lobar pneumonia. Neither patient had gastrointestinal symptoms. Campylobacter bacteremia was diagnosed in both patients. Campylobacter jejuni was identified in the first patient and Campylobacter fetus in the second. Both patients were treated with broad-spectrum antibiotics and recovered fully. PMID- 10482028 TI - Favourable effect of chemotherapy on clinical symptoms and human herpesvirus-8 DNA load in a patient with Kaposi's sarcoma presenting with fever and anemia. AB - The case of a patient infected with human immunodeficiency virus type 1 (HIV-1) with Kaposi's sarcoma who presented with fever of unknown origin, severe anemia, thrombocytopenia and hypoalbuminemia but only limited involvement of the skin is presented. Chemotherapy directed at Kaposi's sarcoma resulted in resolution of these clinical signs and symptoms and was associated with a significant reduction in human herpesvirus-8 DNA load in serum, despite continued HIV-1 replication. Such a decreasing human herpesvirus-8 load following Kaposi's sarcoma-directed chemotherapy has not been reported previously. These findings suggest that Kaposi's sarcoma was indeed responsible for the clinical syndrome and that this neoplasm is a source of human herpesvirus-8 virus particle production, which can be inhibited by chemotherapy-induced reduction in tumor burden. PMID- 10482029 TI - Invasive disease caused by ciprofloxacin-resistant uropathogenic Escherichia coli. AB - To evaluate the invasiveness of ciprofloxacin-resistant Escherichia coli isolated from the urinary tract, the susceptibility to ciprofloxacin of Escherichia coli strains from patients with invasive urinary tract infection was compared with that of isolates from patients with noninvasive disease. In a 14-month period, 2054 different isolates of Escherichia coli were analyzed, of which 554 (27%) were resistant to ciprofloxacin. One hundred twelve (5.4%) strains were isolated from patients with invasive disease. Resistance was significantly less frequent in isolates from patients with invasive disease (4.5%) than in isolates from patients with noninvasive disease (28.3%) (OR, 0.12; CI 95%, 0.05-0.29; P<0.001). Most ciprofloxacin-resistant strains associated with invasive disease were isolated from bacteremic patients who had recently undergone an invasive procedure involving the urinary tract. Invasive disease is caused more frequently by ciprofloxacin-susceptible strains of Escherichia coli, suggesting that resistance to ciprofloxacin may decrease the invasiveness of uropathogenic Escherichia coli. PMID- 10482030 TI - Review of 17 cases of pneumonia caused by Streptococcus pyogenes. AB - Streptococcus pyogenes is an uncommon cause of community-acquired pneumonia and there have been few recent specific accounts of the condition. To describe the current nature of this disease in the UK, data was gathered on patients with clinical pneumonia from whom Streptococcus pyogenes was cultured principally from blood or other relevant normally sterile sites. In the Harrogate and Northallerton districts of North Yorkshire, pneumonia accounted for nine (20%) cases and a quarter of all deaths in a complete sequence of 45 patients with Streptococcus pyogenes bacteraemia detected during the 16-year-period 1981-1996. An analysis is presented of those cases together with eight recent cases from counties York, Durham and Isle of Wight during 1995-1997. Of the total 17 cases, nine occurred in women and eight in men; the age range was 30-92 years. The organism was isolated from blood culture in 15 (88%) patients. Eight (47%) patients died, five within 1 day of hospitalisation. Fourteen (82%) cases occurred in the winter months October to March, including all the fatal cases and all eight in which a clinical 'viral' prodrome was observed. Predisposing medical or surgical conditions were present in 65% of the patients. Major complications included septicaemia, pleural reaction, shock, pulmonary cavitation, osteomyelitis and metastatic abscesses. Seven serotypes of Streptococcus pyogenes were encountered, with M-type 1 predominating (the cause in 60% of cases). All infections were community acquired; two small clusters of fatal pneumonia were seen. PMID- 10482031 TI - Factors related to the prescription of antibiotics for young children with viral pharyngitis by general practitioners and paediatricians in southeastern France. AB - This study evaluates the antibiotic-prescribing practices of physicians as well as other related issues in the context of viral pharyngitis. In a telephone interview, 535 physicians practising in southeastern France were submitted a clinical case description of an episode of acute pharyngitis in a 2-year-old child. Questions concerned antibiotic treatment and physicians' reasons for their treatment decision. The viral origin of the pharyngitis was more likely to be suspected by paediatricians than by general practitioners (92% vs. 78%, P<0.01); 57% of allopaths (compared with 26% of homeopaths/acupuncturists and 14% of paediatricians, P<0.001) declared they would prescribe an antibiotic in this situation (amoxicillin only in 42% of cases). This difference between allopaths and other physicians was still significant after controlling for knowledge regarding antibiotic therapy. In order to limit the risk of emerging resistant bacteria, it is urgent that training be upgraded for physicians, especially for allopaths. PMID- 10482032 TI - Incidence of Borrelia garinii and Borrelia afzelii in Ixodes ricinus ticks in an urban environment, Prague, Czech Republic, between 1995 and 1998. AB - The presence of Borrelia burgdorferi sensu lato and its individual genospecies was determined using a single-step polymerase chain reaction in a total of 779 Ixodes ricinus ticks collected from an urban park in Prague, Czech Republic. The incidence of Borrelia burgdorferi sensu lato was 9.2% in 1995, 3.4% in 1996, 4.5% in 1997 and 2.8% in 1998. The Borrelia garinii to Borrelia afzelii ratio was 1.4:1 and it did not differ significantly throughout the study period. Borrelia burgdorferi sensu stricto was not detected. Borrelia garinii/Borrelia afzelii coinfection was found in 5.7% of positive ticks. The results indicate it is possible for the urban population to come in contact with the causative agent of Lyme disease in an urban park. PMID- 10482033 TI - Microbiological characterization and clinical significance of Corynebacterium amycolatum strains. AB - The laboratory records of patients with bacillus isolates identified as Corynebacterium xerosis were reviewed in an attempt to establish the clinical significance of the isolates, and the isolated strains were reidentified. Of the 22 strains available for reidentification, four were identified as Corynebacterium striatum and 18 as Corynebacterium amycolatum. Forty-one patients were considered to have Corynebacterium amycolatum isolates, and in 13 (31.7%) of these patients a genuine clinical infection occurred, comprising catheter-related infection in seven cases, surgical wound infection in five cases, and pilonidal cyst infection in one case. Most patients were treated with antimicrobial agents (vancomycin in seven cases and amoxicillin/clavulanic acid in four cases). All patients were cured. Corynebacterium amycolatum can cause genuine infection, usually hospital-acquired, and the clinical significance of isolates must be determined to ensure proper management of patients. PMID- 10482034 TI - Influence of increased macrolide consumption on macrolide resistance of common respiratory pathogens. AB - The impact of increased macrolide consumption on the resistance of common respiratory pathogens to erythromycin and azithromycin was evaluated. The study focused mainly on azithromycin. During the period from 1991 to 1996, a 3.5-fold increase in macrolide prescriptions for outpatients was observed in Slovenia. Compared to 1994, when no macrolide resistance was evident in Streptococcus pyogenes and noninvasive Streptococcus pneumoniae, a significant increase in macrolide resistance was observed in these two pathogens in 1997. Moraxella catarrhalis remained uniformly susceptible to macrolides. Close monitoring of macrolide resistance of common respiratory pathogens is thus necessary. PMID- 10482035 TI - Successful treatment of human fascioliasis with triclabendazole. PMID- 10482036 TI - Effect of administering oral and inactivated polio vaccines immediately after birth. PMID- 10482037 TI - Susceptibility of 302 methicillin-resistant Staphylococcus aureus isolates from 20 European university hospitals to vancomycin and alternative antistaphylococcal compounds. SENTRY Participants Group. PMID- 10482038 TI - Staphylococcus cohnii and Ureaplasma urealyticum in a neonate. PMID- 10482039 TI - The effect of testicular macrophages, macrophage-conditioned medium and interleukin-1alpha on the cytoskeleton of bank vole Leydig cells. AB - Recently, morphological and functional interactions between cytoskeletal elements and their involvement in cell movements, shape changes and/or translocation of organelles have been intensively studied. Thus, the aim of our work was to determine whether testicular macrophages and/or their products have an influence on Leydig cell cytoskeleton. The source of Leydig cells and macrophages were male bank voles from spring and autumn generations, reared in different regime of light for 7-8 weeks. The Leydig cells were growing in monocultures or in co cultures with testicular macrophages. All cell cultures were divided to controls or human chorionic gonadotropin-stimulated ones. To some of the cultures testicular macrophage-conditioned medium or interleukin-1alpha were added. The cells were analysed immunocytochemically and radioimmunologically. In Leydig cells obtained from animals kept in a long day, grown in co-cultures with macrophages as well as in those stimulated by testicular macrophage-conditioned medium, distinct rearrangements of microtubules and microfilaments were observed. This phenomenon was strengthened in the presence of hCG in culture media. Concomitantly, basal and hCG-stimulated level of testosterone was enhanced, which indicates the possible involvement of the cytoskeleton in the process of androgen biosynthesis. The influence of IL-1alpha on reorganization of cytoskeletal structures was not observed, suggesting that in the modulation of steroidogenesis by this cytokine cytoskeletal elements do not play an important role. PMID- 10482040 TI - Novel effects of deoxycorticosterone on testicular 11beta-hydroxysteroid dehydrogenase activity and plasma testosterone levels in normal and adrenalectomized rats. AB - The 11beta-hydroxysteroid dehydrogenase (11beta-HSD) protects the testis from the inhibitory effects of corticosterone on testosterone (T) production. The objectives of the present studies were to determine the effects of deoxycorticosterone (DOC) and its mechanism of actions on testicular 11beta-HSD activity and plasma T levels after 7 days of treatment. The results revealed that at the end of 7 days treatment, DOC significantly increased testicular 11beta-HSD activity and plasma T levels in normal rats. However, the time course showed that high plasma T levels lowered 11beta-HSD activity on day 14 and by 21 days both the levels normalized. In adrenalectomized (ADX) rats, only the enzyme activity increased significantly but not plasma T levels. Spironolactone, a competitive inhibitor of mineralocorticoid receptor (MR), did not change testicular 11beta HSD activity in both normal and DOC treated rats suggesting that DOC did not act through MR in increasing 11beta-HSD activity. On the other hand, spironolactone significantly decreased plasma T levels in DOC treated rats. Progesterone (P), a competitive inhibitor of glucocorticoid receptors (GR) or corticosterone significantly suppressed testicular enzyme activity and plasma T levels in DOC treated normal rats. Carbenoxolone which is an inhibitor of 11beta-HSD activity significantly depressed testicular 11beta-HSD activity and plasma T levels in DOC treated normal rats. This paper suggests that DOC increased testicular 11beta-HSD activity through GR; whilst increase in plasma T levels required functioning adrenal glands. The testicular 11beta-HSD is one of the regulators of T levels and vice versa. PMID- 10482041 TI - Coincidence of hot thyroid nodules and primary hyperparathyroidism. AB - Hyperthyroidism is frequently associated with hypercalcemia, which usually subsides after successful treatment of hyperthyroidism. Moreover, thyroid nodules are frequently detected by preoperative thyroid ultrasound in patients with primary hyperparathyroidism. Sensitised by the observation of a patient with coexisting hyperthyroidism and hyperparathyroidism we prospectively evaluated thyroid nodules in euthyroid patients with hyperparathyroidism by thyroid scintigraphy. Whereas the first patient with hyperparathyroidism was hyperthyroid the subsequent four patients with hyperparathyroidism and thyroid nodules had normal fT3 and fT4. Two patients had hypercalcemia and nephroureterolithiasis. Three patients suffered from hypercalcemia and bone pain due to osteoporosis. In the hyperthyroid patient hypercalcemia persisted after euthyroidism was achieved intact parathyroid hormone was found to be elevated. Subsequently, thyroid nodules, detected by preoperative ultrasound in four euthyroid patients with primary hyperparathyroidism, were identified as compensated hot nodules by thyroid scintigraphy. All patients underwent combined subtotal thyroidectomy and parathyroid resection. Histology showed hyperplastic parathyroid glands in one patient and a single parathyroid adenoma in four cases. Postoperatively calcium and PTH levels returned to normal and TSH levels increased in all patients. Persistence of hypercalcemia after successful treatment of hyperthyroidism should be reason for the determination of parathyroid hormone. Thyroid nodules detected by preoperative ultrasound in patients with hyperparathyroidism living in areas of iodine deficiency should be further evaluated by scintigraphy even if TSH is normal. In the case of hot thyroid nodules both parathyroid and partial thyroid resection should be performed. PMID- 10482042 TI - Increased glomerular cytosolic phospholipase A2 activity of OLETF rats with early diabetes. AB - In view of the potential role of prostaglandins (PGs) in development of glomerular hyperfiltration leading to diabetic nephropathy, we studied the temporal relationship of the activity of cytosolic phospholipase A2 (cPLA2), a rate-limiting enzyme for eicosanoid biosynthesis, with hyperfiltration and the histological changes in glomeruli using OLETF rats, a model for non-insulin dependent diabetes mellitus (NIDDM). Diabetes mellitus and associated histopathological changes, which developed spontaneously by 30-46 weeks after birth of OLETF rats, were accompanied by approximately 65% increase in glomerular cPLA2 activity that showed significant correlations with elevated plasma glucose levels and creatinine clearance. Moreover, mesangial cells cultured for 5 days with high glucose exhibited approximately 2-fold higher cPLA2 activity than those cultured with physiologic level of glucose. These data suggest that increased glomerular cPLA2 activity leads to production of PGs, which may promote the progression of early diabetic glomerular hyperfiltration and subsequent diabetic nephropathy. PMID- 10482043 TI - Scintigraphically assessed cardiac sympathetic dysinnervation in poorly controlled type 1 diabetes mellitus: one-year follow-up with improved metabolic control. AB - Diabetic neuropathy is thought to comprise a reversible metabolic and an irreversible structural component of neuronal abnormality. To study whether cardiac sympathetic dysinnervation in poorly controlled longer-term Type 1 Diabetes Mellitus (DM) without myocardial perfusion abnormalities is partially reversible with improved metabolic control, 123-I-metaiodobenzylguanidine (123-I MIBG) scintigraphy (myocardial uptake score 1-6) was performed in 11 Type 1 DM patients (HbA1c 12.0 +/- 1.8%, duration of diabetes 10 +/- 4 yrs) one year after initial assessment. During follow-up, all patients had been treated with intensive insulin therapy and at one year, HbAlc had fallen to 8.4 +/- 1.4% (p < 0.01). The global myocardial 123-I-MIBG uptake score had improved in 5 patients at one year, remained unchanged in 5 patients, and deteriorated in 1 patient. Cardiac sympathetic dysinnervation (123-I-MIBG myocardial uptake (MU) score >2), initially observed in 10 patients, was detectable in 8 patients at follow-up. Myocardial uptake scores of the anterior, lateral, posterior, septal and apical region had improved in 6, 6, 6, 7 and 6 patients, but the mean changes of these scores did not reach significance. In patients with substantial improvement of metabolic control (HbAlc 7.3 +/- 0.6% at one year, mean HbA1c of months 2-12: 7.8%, n = 6), global myocardial uptake had improved from 4.3 +/- 1.0 to 3.2 +/- 1.0 (p < 0.05). Respectively, myocardial uptake score of the anterior, posterior and septal region had ameliorated: 3.8 +/- 1.3 vs 2.3 +/- 0.8 (p < 0.05), 4.0 +/- 1.7 vs 2.5 +/- 1.0 (p < 0.05), 4.3 +/- 1.2 vs 3.2 +/- 1.5 (p < 0.05). In conclusion, cardiac sympathetic dysinnervation in poorly controlled longer-term Type 1 DM patients is dominated by irreversible neuronal abnormalities. Substantial metabolic improvement, however, partially restores cardiac sympathetic dysinnervation, indicating the presence of a reversible component of cardiac sympathetic dysfunction in longer-term Type 1 DM. PMID- 10482044 TI - Measurement of insulin sensitivity: influence of potassium supply during euglycaemic glucose clamps in healthy volunteers. AB - Insulin sensitivity can be quantitatively measured by the hyperinsulinaemic euglycaemic glucose clamp technique. Infusion of insulin during the clamp procedure leads to a decline of kalaemia unless potassium is supplied. We investigated whether supplementation of potassium during euglycaemic glucose clamps influences insulin sensitivity. In a randomised study the insulin sensitivity index (S(I)) was determined with two-step hyperinsulinaemic (insulin infusion rates 0.25 (step 1) and 1.0 mU kg(-1) min(-1) (step 2)) euglycaemic (5.0 mmol L(-1)) glucose clamps in 20 healthy male volunteers on two different study days. On one day blood potassium was kept constant by means of a variable i.v. potassium chloride infusion ("eukalaemic potassium clamp"), whereas on the other day the decline in blood potassium was monitored only. Without potassium supply kalaemia decreased from basal levels of 4.35 +/- 0.18 mval L(-1) to 4.25 +/- 0.17 (step 1) and further to 3.88 +/-0.14 mval L(-1) (step 2 (mean +/- SD)). Without and with potassium supply the insulin sensitivity index measured was comparable (S1 10.6 +/- 3.6 vs. 9.5 +/- 3.5 ml min(-1) m2 per microU ml(-1), n.s.; glucose infusion rates 3.6+/-1.6/12.6 +/- 2.6 (step 1/step 2) vs. 3.7 +/- 1.5/12.2 +/- 2.7 mg kg(-1) min(-1), n.s.). In conclusion, this study shows that potassium supply during hyperinsulinaemic euglycaemic glucose clamps in healthy subjects does not influence the insulin sensitivity index. PMID- 10482045 TI - Amino acid polymorphism Gly 972 Arg in IRS-1 is not associated to lower clamp derived insulin sensitivity in young healthy first degree relatives of patients with type 2 diabetes. AB - The Gly 972 Arg variant in the insulin receptor substrate-1 (IRS-1) gene may interact with the pathogenesis of common insulin-resistance disorders raising the hypothesis that the mutation may predispose to type 2 diabetes. We examined the codon 972 variant in 144 non-diabetic first degree relatives of patients with type 2 diabetes (FDR), who underwent extensive phenotyping: Glucose tolerance was determined by an oral glucose load, insulin sensitivity by euglycaemic hyperinsulinaemic glucose clamp (glucose metabolic clearance rate, MCR) and body composition by bioelectrical impedance. 20 (14%) of the FDR showed the Gly 972 Arg variant in heterozygous form, 2 (1.3%) probands were homozygous. Carriers of the polymorphism did not differ in MCR independent of body weight and total body fat. The polymorphism does not seem to determine clamp-derived insulin sensitivity. Despite identical fasting plasma glucose, carriers of the polymorphism showed a slightly lower fasting serum insulin and lower insulin response to an oral glucose load but higher glucose concentrations. In an obese subgroup (BMI > 25) the polymorphism did not show a higher frequency and was not associated with lower insulin sensitivity. In the investigated group of young, healthy relatives of type 2 diabetes patients, the frequency of the mutation corresponded to that of a diabetic population. In summary our data show that the polymorphism is not suitable to predict insulin resistance. PMID- 10482046 TI - The prevalence of IDDM in the first degree relatives of children newly diagnosed with IDDM in Austria--a population-based study. Austrian Diabetes Incidence Study Group. AB - Insulin dependent diabetes mellitus shows a strong familial predisposition and an unexplained geographical variation in incidence. It is not known whether the risk of IDDM in first degree relatives depends on the risk in the background population. The aim of the study was to assess the prevalence of IDDM in parents and siblings of newly diagnosed children with IDDM in Austria, a known area of low risk for IDDM. The family history data of all diabetic children (< 15 years) diagnosed between 1988-1994 in Austria were analysed. The cumulative incidence of IDDM in siblings of newly diagnosed diabetic children was 0.0026772 cases/year, this means 29.7 times increased risk compared to the background population. Of the diabetic children 5.8% had at least one parent with IDDM and the prevalence of IDDM in fathers (3.9%) was higher (p = 0.015) compared to mothers (1.9%). The risk of IDDM tended to be higher for offsprings of diabetic fathers (OR 3.8, p < 0.003) in families with 2 or more children than in single child families, where the prevalence was 4.2% both in fathers and mothers. In conclusion the prevalence of IDDM in parents of diabetic children in Austria was lower than reported in populations with high IDDM incidence. This may reflect a lesser degree of genetic predisposition of the Austrian population. The prevalence of IDDM in siblings was similar to that in high risk populations. We saw an interaction of gender of the diabetic parent and diabetic offspring and the family size. PMID- 10482047 TI - Anti-neutrophil cell antibodies in newly diagnosed patients with type-1-diabetes. AB - Both anti neutrophil cell antibodies and anti endothelial cell antibodies were found in 7 out of 30 newly-diagnosed type-1 diabetic patients. This confirms the abnormal activation of the immunological system in the early stage of type-1 diabetes mellitus. PMID- 10482048 TI - Non-tuberculous mycobacterial tenosynovitis: a review. AB - The clinical characteristics, outcome and treatment of non-tuberculous mycobacterial tenosynovitis are reviewed. From lesions localized in the hand, 10 different species of non-tuberculous mycobacteria have been reported. The most common are Mycobacterium marinum and Mycobacterium kansasii. Other less frequent organisms are Mycobacterium avium complex, Mycobacterium szulgai, Mycobacterium terrae, Mycobacterium fortuitum, Mycobacterium chelonae, Mycobacterium abscessus, Mycobacterium malmoense and Mycobacterium xenopi. The infections appear to be the result of previous trauma, surgical procedure, corticosteroid injection or non apparent inoculation (water contamination). Immunosuppression is sometimes associated with the infections and can be considered as a risk factor. Surgical debridement and appropriate mycobacterial cultures are critical to enable diagnosis and appropriate management. Specimens should be inoculated on a range of media and incubated at a range of temperatures in order to isolate mycobacteria with different growth characteristics (with prolonged incubation). The optimal treatment of these infections is discussed. PMID- 10482049 TI - C-reactive protein levels in HIV complicated by opportunistic infections and infections with common bacterial pathogens. AB - In order to determine the pattern of C-reactive protein (CRP) concentrations in HIV-infected patients with various other infections, we conducted a prospective study (for the period 1990-91) of all HIV-seropositive patients hospitalized with fever and a retrospective study (for the period 1990-95) of all patients infected with Mycobacterium avium complex (MAC) and Pneumocystis carinii pneumonia (PCP). Samples from blood, cerebrospinal fluid and sites with clinical signs of infection were obtained for bacteriological culture. Polymerase chain reaction (PCR) determination was performed for cytomegalovirus in blood and CSF. Patients with opportunistic infections had a significantly lower increase in CRP concentration than patients infected with common bacterial pathogens. Patients with PCP and mycobacterial infections had a distinct CRP response after the onset of therapy. Lack of CRP increase at diagnosis of MAC infection was associated with a shorter survival and normalization of CRP after MAC therapy with a significantly longer survival. PMID- 10482050 TI - Identification of human immunodeficiency virus type 1 subtypes B and F B/F recombinant and dual infection with these subtypes in Argentina. AB - DNA sequences encoding the third variable region (V3) of human immunodeficiency virus type-1 (HIV-1) envelope glycoprotein gp120 were obtained from 18 infected individuals residing in different regions of Argentina. Proviral DNA representing the env V3 region was obtained by PCR from uncultured peripheral blood mononuclear cells (PBMC) and genetic heterogeneity was examined by phylogenetic analysis. Sequences representing the gag p17 region were also obtained for a subset of these samples. Moreover, 1 sample that it was not possible to classify according to initial phylogenetic analysis was further analysed by molecular cloning of both V3 and p17 regions. Phylogenetic analysis according to different methodologies were performed comparing obtained sequences with a set of reference sequences representing previously characterized HIV-1 subtypes. The recombinant identification program (RIP) was used to study the presence of possible recombinant sequences. Phylogenetic analysis demonstrated that viruses representing both subtypes B and F are circulating among HIV-1 infected individuals in Argentina. In addition, RIP analysis showed that an initially unclassified sequence exhibited similarities to subtypes B and F in different fragments of the V3 region. Separate phylogenetic analysis of each of these fragments revealed divergent clustering, suggesting that this sequence harbours a point of recombination within the V3 loop. Interestingly, we also identified a dually infected individual with viruses belonging to subtypes B and F, as demonstrated by molecular cloning analysis of the env V3 and the gag p17 regions. Taken together, our study shows that both subtypes B and F are circulating in different regions of Argentina. Moreover, the data presented here show that dual infections with subtypes B and F can occur, and consequently B/F recombinant sequences are arising in the region. PMID- 10482051 TI - Transmission of a minor variant of transcriptionally active HIV-1. AB - We investigated the genotypic variation of pro-viral human immunodeficiency virus type 1 (HIV-1) DNA in a virus donor-recipient pair by comparing sequences from the HIV-1 V3 region of the gp120 gene and the p17gag gene. We found that the transmitted virus was a minor variant in the donor's virus population in blood. Possible selection mechanisms within the host (neutralization by antibodies) were studied in order to investigate whether antibodies could explain the conserved HIV genotype found in the recipient. In conclusion, our data indicate that a minor variant of pro-viral transcriptionally active HIV-1 found in PBMC was transmitted from donor to recipient. Development of a homogeneous genotype regarding the V3 region (V3d-B1(1)) of pro-viral DNA in the recipient's PBMC and a partially homogeneous genotype regarding the HIV-RNA was possibly caused by an envelope associated selection that is not dependent upon neutralizing antibodies. PMID- 10482052 TI - Changing epidemiology of hepatitis B infection in Finland. AB - Finland has, until recently, had a very low incidence of hepatitis B infection, reflected in transmission mainly amongst adolescent and adult age-groups with high-risk behaviour. Several recent local outbreaks of acute hepatitis B in Finland may indicate a change in the epidemiology of this infection. We examine time trends of hepatitis B notifications to the new national infectious disease registry. We also analyse all hepatitis B cases notified over the 2-y period from 1 January 1995 to 31 December 1997 by age, sex, place of residence, country of birth and reported route of transmission. The reported incidence of acute hepatitis B increased 3-fold over this period. The main reported route of transmission was related to injecting drug use, although a significant proportion of adolescent female cases was reportedly infected through heterosexual intercourse. A critical evaluation of the current targeted vaccination approach and other prevention policies is required. PMID- 10482053 TI - Chlamydia pneumoniae is an important cause of community-acquired pneumonia in school-aged children: serological results of a prospective, population-based study. AB - The aetiology of community-acquired pneumonia in childhood was studied in the total population of 8851 children in the area of 4 municipalities in eastern Finland. All cases of community-acquired pneumonia (n = 201) were registered during a surveillance period of 12 months between September 1, 1981 and August 31, 1982. The diagnosis of pneumonia was verified radiologically in all identified cases. The diagnosis of chlamydial infection was based on an antibody response measured by complement fixation (CF), by enzyme immunoassay (EIA; IgG or IgM) or by microimmunofluorescence (MIF; IgG or IgM), and the diagnosis of mycoplasmal infection on CF alone. In total, 29 cases of Chlamydia sp. infection were diagnosed; 20 were caused by Chlamydia pneumoniae. Thus, C. pneumoniae was an aetiological agent in 10%, of the 201 pneumonia cases: the proportion was 9% for children aged 5-9 y and 31% for those aged 10 y or more. In the study population, the total incidence of C. pneumoniae pneumonia was 2.3/1000/y. Mycoplasma pneumoniae serology (CF) was positive in 44 patients (22%); the total incidence of M. pneumoniae pneumonia was 5.0/1000/y. Serological evidence of both Chlamydiae and M. pneumoniae was detected in 9 (41%) patients. Our results indicate that C. pneumoniae is an important cause of community-acquired pneumonia in school-aged children. Diagnostic serological response to Chlamydia species or M. pneumoniae was found in 42% of pneumonia patients between 5 and 9 y of age and in 67% of patients aged 10 y or more. Thus, we suggest that macrolides should be considered as an empirical antimicrobial treatment for community-acquired pneumonia, especially in school-aged outpatients. PMID- 10482054 TI - No serological evidence of association between chlamydia pneumonia infection and acute coronary heart disease. AB - Today's medical literature shows more and more evidence that Chlamydia pneumoniae plays a role in coronary atherosclerotic disease. This cross-sectional study examines the serostatus for C. pneumoniae of patients with an acute cardiovascular event. A total of 58 patients with acute myocardial infarction or with proven unstable angina (occlusion > or = 70% of at least 1 vessel at coronary angiography) ('patients') were compared with 58 age- and sex-matched patients without a cardiovascular event ('controls'). Serological testing for C. pneumoniae was performed by a microimmunofluorescence test during the first week of the event and 4-6 weeks later. Although more patients from the cardiovascular event group showed a positive history of a respiratory tract infection during 6 weeks preceding hospitalization (18/58 patients, 8/58 controls, p = 0.03), there was no significant difference between the two groups for an acute, chronic or past infection by C. pneumoniae (19/58 patients, 24/58 controls, p = 0.93). In conclusion, contrary to previous published papers, this study could not confirm an association of C. pneumoniae infection with an acute coronary event. Serological testing alone may not be the best way to demonstrate this association. An acute infection by C. pneumoniae, which should have been detected by serological testing, is probably not the origin of the rupture of an atheromatous plaque. PMID- 10482055 TI - Evolution of serum beta2-microglobulin concentrations during treatment of tuberculosis patients. AB - A total of 36 human immunodeficiency virus-seronegative patients were studied in order to evaluate serum beta2-M levels in immunocompetent patients with tuberculosis who were receiving treatment. Six measurements of several clinical and laboratory parameters were carried out at different intervals during the 6 months of treatment. The mean serum beta2-M at presentation was 149 nmol/l and 4 patients had values above the upper normal limit. Significant decreases in the mean serum beta2-M concentration were observed in the follow-up determinations in the patients as a whole (p = 0.002), in the patients with normal (p = 0.039) and in the patients with increased beta2-M at presentation (p = 0.037). beta2-M significantly correlated with erythrocyte sedimentation rate (p = 0.002). The statistically significant decrease observed in patients with both normal and increased beta2-M values at presentation, suggests that the immunological dysfunction responsible for the increase in beta2-M involves most, if not all, patients with tuberculosis. The measurement of beta2-M in conjunction with other clinical and laboratory parameters could be helpful in evaluating the response to therapy, particularly in those patients with increased beta2-M at presentation. PMID- 10482056 TI - Line probe assay in the rapid detection of rifampin-resistant Mycobacterium tuberculosis directly from clinical specimens. AB - The sensitivity of a PCR-based line probe assay (Inno-LiPA Rif. TB Assay; Innogenetics NV Zwijndrecht, Belgium) was studied by using nested-PCR technique. A total of 75 specimens, representing various body locations from 70 suspected tuberculosis patients were obtained. LiPA yielded 30 Mycobacterium tuberculosis complex positive results (sensitivity 58.8%, compared with final diagnoses) whereas culture for M. tuberculosis was positive in 18 specimens (sensitivity 35.3%). Genotypic rifampin resistance testing by LiPA showed that 7 specimens contained rpoB mutations associated with RMP resistance, and sequencing data of the rpoB gene and LiPA patterns agreed in 29 of 30 M. tuberculosis positive specimens (96.7%). This indicates reliable performance, which makes the test suitable for the rapid determination of resistance to rifampin directly in clinical samples. However, the best results are obtained if LiPA is combined with conventional staining and culture methods. PMID- 10482057 TI - Helicobacter pylori in the natural environment. AB - The presence of Helicobacter pylori (H. pylori) in the natural environment has been demonstrated in a number of studies. However, its route of infection into humans is unknown. To study this further, we attempted to detect H. pylori in the natural environment in a region of Japan with a high infection rate. Tap and well water and field soil samples were collected from around the residences of subjects who had participated in an epidemiological survey in 1996. Samples of water from rivers and ponds, and specimens of flies and cow faeces were collected in the region. DNA was extracted from the water, field soil and faecal samples after selective collection of H. pylori by the immunomagnetic-bead separation technique. H. pylori-specific DNA was detected in water, field soil, flies and cow faeces by nested polymerase chain reaction (PCR), and the ureA partial sequences of the PCR products were aligned. The nucleotide sequences of the samples amplified by PCR were highly homologous (96-100%) with the H. pylori sequence in the GenBank database and the H. pylori-specific DNA sequences were highly homologous with each other. These findings suggest the existence of H. pylori in the natural environment and a possible transmission route. PMID- 10482058 TI - Bordetella pertussis, Bordetella parapertussis, Mycoplasma pneumoniae, Chlamydia pneumoniae and persistent cough in children. AB - Material collected during a prospective pertussis vaccine trial in 1992-95 was examined for Bordetella pertussis (culture and serology), Bordetella parapertussis (culture), Mycoplasma pneumoniae and Chlamydia pneumoniae (PCR). From 64% (99/155) of episodes with cough for less than 100 d, 115 aetiological agents were identified in one southern and one northern subset of DT-recipients. The most common single agent was B. pertussis, representing 56%(64/115), with a median cough period of 51 d, followed by M. pneumoniae 26%(30/115), 23 d, C. pneumoniae 17% (19/115), 26 d, and B. parapertussis 2% (2/115). For co infections, the median duration of cough was about 60 d. Spasmodic cough for 21 d or more (clinical WHO criteria for pertussis) was present in 82% (41/50) of infections with B. pertussis as single agent, 38% (17/45) with B. parapertussis, 38% (5/13) with C. pneumoniae, 26% (5/19) with M. pneumoniae and 30%(17/56) in cases where no aetiology was found. In children with cough for more than 100 d (n = 78) using all vaccine arms, B. pertussis was responsible in 83% (65/78), in 21%(16/78) together with other agents. Acellular vaccines were more efficient against serious disease than whole cell vaccine. Antibiotic treatment was more common at the southern (34%) study site than at the northern one (12%). The findings indicate that diagnosis should rely on laboratory confirmation, both for rational treatment of an individual case and for monitoring outbreaks. PMID- 10482059 TI - Enterobacteriaceae meningitis in adults: a review of 20 consecutive cases 1977 97. AB - Enterobacteriaceae are not a frequent cause of meningitis in adults and are seen mainly in neurosurgical patients and on occasion in elderly and debilitated patients. Consequently, most series studied have been small and selected. In order to obtain a clearer clinical picture, we reviewed 20 consecutive cases of Enterobacteriaceae meningitis admitted to the Department of Infectious Diseases, Rigshospitalet, Copenhagen, during the years 1977-97. They comprised 1.5% of all cases of acute bacterial meningitis admitted to the department. All of the patients were either elderly and/or had 1 or more underlying diseases and predisposing factors. The clinical presentation and cerebrospinal fluid findings were not different from that of acute bacterial meningitis in general. The mortality rate was 40% and correlated with simultaneous bacteraemia. Complications were seen in a further 30% of patients and 25% survived with different sequelae. These high rates may, at least in part, be due to the advanced age and debilitated state of the patients studied. Escherichia coli was the most frequent of the Enterobacteriaceae. PMID- 10482060 TI - Endemic burkholderia cepacia bacteraemia: clinical features and antimicrobial susceptibilities of isolates. AB - Burkholderia cepacia has emerged as a nosocomial pathogen, causing numerous outbreaks, particularly among cystic fibrosis (CF) patients. Reports of clinical features of endemic B. cepacia bacteraemia in non-CF patients are rare. Twenty five patients with B. cepacia bacteraemia were matched with 25 controls with nosocomial Escherichia coli bacteraemia at China Medical College Hospital, Taichung, Taiwan, over a period of 3 y. Case-patients included 16 men and 9 women, from 13 to 75 y. All had severe underlying diseases, most commonly malignancy (44%). Twenty-four patients (96%) had nosocomial infections. Five patients (20%) had polymicrobial bacteraemia. Our controls included 11 men and 14 women, age range 18-80 y. The most common underlying disease was malignancy (44%). Multivariate analysis revealed that indwelling central venous catheter was the significant risk factor predisposing to B. cepacia bacteraemia (p= 0.025). Eleven case-patients met the definition of catheter-related bloodstream infection. Fifteen patients (60%) received appropriate antimicrobial therapy after notification of positive blood cultures and susceptibility patterns. The overall case-fatality rate was 12% (3/25), only 1 of whom died of B. cepacia bacteraemia. There was no statistically significant difference in overall mortality rate between case-patients and controls. All isolates were susceptible to ceftazidime, piperacillin and minocycline and 84% of the isolates were susceptible to imipenem. B. cepacia should be considered a potential pathogen in hospitalized patients with severe underlying diseases, particularly those with indwelling central venous catheters. PMID- 10482061 TI - Evaluation of pneumonia diagnosis in intensive care patients. AB - The aims of this study were to investigate the frequency of bronchopneumonia diagnosed by histological criteria among autopsied intensive care unit (ICU) patients and to compare these with rates of pneumonia diagnosed by conventional clinical methods. The study material comprised 141 autopsied ICU patients from 7 ICUs in university hospitals in Copenhagen from a 1-y period. A total of 20 lung tissue specimens were sampled from each patient and the histopathological diagnoses were classified as no, mild, moderate or severe bronchopneumonia. Inter observer variation was calculated using kappa statistics. Demographic data and diagnoses of pneumonia were registered from the patient files. Twenty-six percent of the patients had pneumonia diagnosed whilst in the ICU. Histological evidence of pneumonia, found for every second patient, was regarded as the gold standard. Diagnosis of pneumonia in the ICU had a sensitivity of 29% and if diagnoses of pneumonia during the month before ICU-admission were included, a sensitivity of 60% was found. Specificity for pneumonia diagnosed in the ICU was 77%. The percentage of all ICU-patients with pneumonia was calculated to be between 36% and 56%, depending on the extent of excess mortality attributable to pneumonia. Pulmonary segments with histologically diagnosed pneumonic lesions were distributed diffusely, although the upper segments tended to be affected less. Nearly all patients had other histopathological findings than bronchopneumonia. The reliability coefficient among the 6 pathologists was found to be moderately good (kappa = 0.45). PMID- 10482062 TI - Risk factors for Toxoplasma infection in pregnancy: a case-control study in France. AB - Each year an estimated 4900 cases of primary Toxoplasma infection occur in pregnant women in France, a country with a high prevalence. Since 1992 all pregnant women at risk of Toxoplasma infection have been required to undergo monthly serological testing. This case-control study, the first of its kind in France, was undertaken to identify risk factors for Toxoplasma infection during pregnancy, with a view to improving primary prevention among non-immune pregnant women. A total of 80 pregnant women who seroconverted to Toxoplasma were matched with 80 pregnant women who had repeatedly negative tests. The women were interviewed by telephone, using a standardized questionnaire, to determine socio demographic characteristics, exposure to possible risk factors and the type of information on prevention received during pregnancy. The risk factors for Toxoplasma infection included in a multivariate analysis were poor hand hygiene (OR = 9.9; 95%CI: 0.8-125), consumption of undercooked beef (OR = 5.5; 95%CI: 1.1 27), having a pet cat (OR =4.5; 95%CI: 1.0-19.9), frequent consumption of raw vegetables outside the home (OR = 3.1; 95%,CI: 1.2-7.7) and consumption of undercooked lamb (OR = 3.1; 95%CI: 0.85-14). Receipt of documentary advice on prevention was associated with a lower risk of infection. Prevention campaigns among pregnant women in France could be improved and should focus on eating habits, hand hygiene and cats. PMID- 10482063 TI - Teratogenic evaluation of oxacillin. AB - Teratogenic studies of oxacillin in humans have not been published. The population-based data-set of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1996 contains 22,865 foetuses or newborns with congenital abnormalities and 38,151 matched control newborns without congenital abnormalities. The mothers of 14 cases and 19 controls were treated with oral oxacillin during pregnancy. A teratogenic risk of oxacillin was not detected in this case-control study. PMID- 10482064 TI - Human herpes virus 6 fatal encephalitis in a bone marrow recipient. AB - Human herpes virus 6 (HHV-6) has been implicated as a human pathogen in both normal and immunocompromised hosts. It has been associated with interstitial pneumonitis and bone marrow suppression after transplantation. We report here a case of fatal encephalitis in a bone marrow transplant recipient. This case illustrates the importance of considering HHV6 as an emerging pathogen in immunocompromised hosts. PMID- 10482065 TI - Group B streptococcal vertebral osteomyelitis with bacteraemia in an adult with no debilitating condition. AB - A previously healthy 62-year-old immunocompetent woman presented with group B streptococcal vertebral osteomyelitis. Group B Streptococcus was recovered in 3 consecutive blood cultures. The patient recovered fully after treatment including antibiotic therapy, bed rest and physical rehabilitation. Group B streptococcal vertebral osteomyelitis is uncommon and has not previously been reported in patients with no immunosuppressive condition. PMID- 10482066 TI - Bilateral mammary abscess due to Brucella melitensis. AB - Brucellosis is an important zoonosis, which is widespread as endemic cases in Turkey. Unusual clinical features and complications can sometimes lead to diagnostic difficulties. We describe here a case of a pregnant woman with bilateral mammary abscesses due to Brucella melitensis. PMID- 10482067 TI - Primary psoas abscess due to Fusobacterium nucleatum. AB - A case of primary pyogenic psoas abscess due to Fusobacterium nucleatum is described. Clinicians must maintain a high index of clinical suspicion for the diagnosis of psoas abscess. Although Staphylococcus aureus accounts for most cases of primary psoas abscess, this report emphasizes the importance of bacteriological confirmation of the microorganism involved. PMID- 10482068 TI - Immunochemical and molecular characterization of vertebral hydatid fluid. AB - This study describes a 42-year-old male born in Tunisia, who presented with vertebral hydatidosis which was initially misdiagnosed as Pott's disease. The correct diagnosis was finally made utilizing CT and MRI imaging and confirmation of the diagnosis was made possible by molecular and immunochemical characterization of the content of vertebral cyst fluid. PMID- 10482069 TI - Visceral larva migrans syndrome complicated by liver abscess. AB - We describe a case of visceral larva migrans syndrome complicated by liver abscess, pericardial effusion and ascites. To our knowledge, these findings have not been reported previously. The structural and immunological alterations caused by visceral larva migrans are thought to lead to the development of visceral abscesses. PMID- 10482070 TI - Pyogenic infection of the pubic symphysis in a diabetic patient. AB - A 51-year-old woman with diabetes mellitus had swelling in the inguinal area leading to discharge. Insulin, irrigation of the wound and intravenous antibiotics had no effect. Fistulography and a fistula-CT showed a fistula connecting to the pubis symphysis. After curettage of the pubic symphysis, the wound was closed. PMID- 10482071 TI - Chlamydia pneumoniae as a trigger of reactive arthritis. PMID- 10482072 TI - Fungaemia due to thermophilic cryptococci: 3 cases of Cryptococcus laurentii bloodstream infections in cancer patients receiving antifungals. PMID- 10482073 TI - Teaching social inequalities in health: barriers and opportunities. AB - This article examines some of the main threats and new opportunities encountered by teachers of social inequalities in health in contemporary academia. Focusing mostly on the recent US and European experiences, I suggest that lay world views legitimating social inequalities are often in conflict with explanations arising from social epidemiology and medical sociology. The dominance of medicine in public health, through its often implicit assumptions about the biological determinants of human behaviour, is also identified as a barrier to teaching social inequalities in health. Educational elitism, which restricts higher education to members of the upper middle class, is identified as another barrier to teaching social inequalities in health. On the other hand, teachers in this field can benefit from a recent growth of empirical studies during the last decade aimed at understanding the social determinants of health inequalities. Finally, I suggest that familiarity with current critical scholarship within public health, as well as the use of techniques developed by sociologists to teach social stratification, can be valuable resources for teaching social inequalities in health. PMID- 10482074 TI - Genetic information and genetic discrimination how medical records vitiate legal protection. A comparative analysis of international legislation and policies. AB - This study compares currently enacted and pending legislation and policies concerning genetic information and genetic discrimination in Europe and the USA. The emergence of a multi-disciplinary scientific field comprising genetically related areas and the discovery of new genetic factors have improved our ability to predict genetic risks associated with illness. Early detection and preventive healthcare thus hold promise for improving public health, but much remains unknown about the actual risks deriving from genetic testing. Positive (not normal) test results may give little certainty of the actual severity of a disease. Genetic information could thus be used for social stigmatization and genetic discrimination. Western laws have been enacted in a context of unknown potential risks and benefits, and an increased number of regulatory initiatives is to be expected. Yet, there is currently no comprehensive US or European legal approach concerning genetic information and genetic discrimination and, despite legislation thus far enacted, the protection of individuals remains uncertain, and this could have a negative impact on attitudes to genetic testing, with important consequences for public health strategies to prevent disease. PMID- 10482075 TI - Medical practice variations: changing the theoretical approach. AB - Variations in medical practice are well documented, but there has been less progress in explaining these variations. This paper discusses the existing theories and hypotheses and concludes that a change in theoretical approach is required, to one that more directly highlights the social context influencing the behaviour of doctors in their daily practice. An initial alternative model for explaining variation in practice style is presented. The paper illustrates how (combinations of) important structural factors, such as the availability of hospital resources, the way the doctor is reimbursed, the availability of patients, professional uncertainty, and the way the hospital is financed, lead to hypotheses about when different local standards of medical care emerge. It is concluded that theoretical progress in research on variations in medical practice is possible and that empirical research needs to be driven by hypotheses that emphasize the role of social contexts in the doctor's decision behaviour. Some suggestions for future lines of research are outlined. PMID- 10482076 TI - Inequalities in health: evidence from Denmark of the interaction of circumstances and health-related behaviour. AB - It is well known that the experience of poor health depends on adverse social and material circumstances and on unhealthy behaviour. In 1990 a British study asserted that privileged people gain more health benefit from a healthy lifestyle than do deprived people. In the present study this assertion was taken as a hypothesis, assuming statistical interaction between circumstances and health behaviour. The combined effect of these variables was studied in Denmark. Data was obtained from a 1987 national Health and Morbidity Survey; variables were selected to correspond closely with those in the earlier British study. The analysis included a multivariate analysis of variance. The results show that health-related behaviour has a positive effect on health both for those who are deprived and those who are not deprived, which is at variance with the findings of the British study. It confirms earlier analyses of Danish and Dutch data. PMID- 10482077 TI - The influence of social deprivation as measured by the CNI on psychiatric admissions. AB - The purpose of this study was to examine the importance of social deprivation for psychiatric admissions and its correlation with two different deprivation scores. Care Need Index (CNI) and Townsend scores were calculated at the small area level in Malmo, a city in southern Sweden. Admission rates for all psychiatric inpatients from Malmo aged 20-79 years, admitted to the psychiatric and alcohol clinics from 1 January 1991 to 31 December 1994, were calculated. The relationship between the CNI and psychiatric admissions was analysed by applying a Poisson regression model. The results are shown as incidence density ratios (IDR) with 95% confidence intervals (CI). From the most deprived areas, the first psychiatric admission rate was more than four times higher than in the most affluent areas. The rates of second and third admission were even higher. Admissions to the alcohol clinic were similar to psychiatric admissions, but the most deprived areas had first admission rates about ten times higher than in the most affluent areas. About 27% of first admissions, including patients from both psychiatric and alcohol clinics, had a diagnosis of psychosis, and 43% were substance abusers. There were differences between the patients' diagnoses in different areas. The correlation between the CNI and Townsend scores was very high. The most important finding of this study is the strong correlation between social deprivation, based on different deprivation indices, and first admissions to psychiatric and alcohol clinics. PMID- 10482078 TI - Marital disruption and long-term work disability. A four-year prospective study. AB - Data from a community-based four-year prospective study were used to investigate the relationship between marital disruption and long-term work disability. In 1990, all inhabitants of the municipality Ullensaker, Norway, belonging to six cohorts (20-22 years, 30-32 years, 40-42 years, 50-52 years, 60-62 years and 70 72 years) were sent a questionnaire. Of the 1,359 respondents who were working, married or cohabiting, and not older than 62 years, 1,115 (82%) returned a second questionnaire four years later (1994). Separation or divorce between 1990 and 1994 was related to long-term work disability in 1994 (OR = 3.02; 95% CI: 1.57 5.81), even after adjustments for age, sex, work characteristics, number of work hours per week, job satisfaction, body mass index, having pre-school children, smoking, physical leisure activity, emotional symptoms and musculoskeletal pain (all measured in 1990). Emotional problems evoked by the marital disruption may be part of the explanation. PMID- 10482079 TI - The link between socioeconomic status and breast cancer--a possible explanation. AB - The aim of this ecological study was to explore possible associations between childhood nutrition and breast cancer. The study compared heights and weights of children of different socioeconomic status and subsequent breast cancer risk in Queensland, Australia. In 1950 12-year-old girls from families of high socioeconomic status were 3 cm taller and 6.6 kg heavier than girls of the same age of lower socioeconomic status. Approximately 35 years later, age standardized mortality rates for breast cancer among all Queensland women were approximately 10% higher for women of high compared with low socioeconomic status. Taking into account the limitations of ecological studies and risk factors other than nutrition, these results are compatible with the hypothesis that there is an association between childhood nutritional experiences and subsequent risk of breast cancer. PMID- 10482081 TI - Population density and perinatal mortality in Norway and Sweden 1975-1988. AB - Data from the medical birth registries in Norway and Sweden were used to study geographical variations in perinatal mortality in the two countries. The study population comprised 1.4 million single births during 1975-79 and 1985-88. Perinatal death (n=9,834) for infants with a birthweight of 1,000 g or more was the main measure of outcome. In both periods and in both countries, perinatal mortality was inversely associated with population density and annual number of births, with relative risks amounting to 1.4 in Norway and 1.2 in Sweden. Among live births, these gradients were observed in deaths from asphyxia or immaturity in the first time period, and in deaths from congenital malformations in the second time period. As care for premature infants and infants with respiratory problems improves, geographical variations in perinatal mortality may to a larger extent depend on differences in medical care of infants affected by congenital malformations as well as differences in efforts to reduce the prevalence at birth of serious congenital malformations. PMID- 10482080 TI - Lessons from the Oslo Study Diet and Anti-smoking Trial: a qualitative study of long-term behaviour change. AB - The aim of this study was to identify factors associated with sustainable behaviour change among persons at increased risk for future cardiovascular disease. A qualitative approach based on focus group interviews was used. Twenty five men aged 62-71 years who participated in the Oslo Study Diet and Anti smoking Trial were interviewed in four focus groups. The study identified five categories of factors linked to the process of lifestyle changes: the doctor patient relationship, significant others, motivators, barriers, and empowerment. A strong personal attachment to the principal investigator of the Oslo Study was revealed. The relationship was described as friendly and supportive. The family based approach and the impact of spouses, in particular, was emphasized by the participants. Somatic symptoms and fear often acted as motivators to behavioural change. Individual feedback was also identified as an important motivator. Psychological resistance to external pressure, concerns regarding behaviour change, and benefits from current (unhealthy) behaviour were identified as barriers. Participants reported a process of empowerment and personal control as an important element of sustainable change. The importance of the unique relationship with a certain provider has implications for future healthcare planning. This factor may also limit the external validity of studies of human behaviour. PMID- 10482082 TI - Has increased focus on vocational rehabilitation led to an increase in young employees' return to work after work-related disorders? AB - The aim of this study was to determine whether the large investments in vocational rehabilitation made in Sweden during the 1990s had improved the level of return to work for young employees and to study the factors predicting return to work. The study population comprised all employees under 30 years of age whose reports on work-related disorders were under consideration at regional social insurance offices in Vasterbotten county in 1990 and 1994 (n = 266). Between these years, increased efforts were made by the Swedish government to improve vocational rehabilitation. Data was collected from the register and by means of questionnaires. It was found that employees with musculoskeletal disorders were more likely to return to work during periods of intensive vocational rehabilitation. No increase in the level of return to work was apparent if all disorders were considered. Men showed a higher level of return to work than women, although women were better educated. PMID- 10482083 TI - Validity and reliability of Resource Utilization Groups (RUG-III) in Finnish long term care facilities. AB - Resource Utilization Groups, Version III (RUG-III) is a case-mix system developed in the USA for classification of long-term care residents. This paper examines the validity and reliability of an adapted 22-group version of RUG-III (RUG III/22) for use in long-term care facilities in Finland. Finnish cost weights for RUG-III/22 groups are calculated and different methods for their computation are evaluated. The study sample (1,964 residents) was collected in 1995-96 from ten long-term care facilities in Finland. RUG-III/22 alone explained 38.2% of the variance of total patient-specific (nursing + auxiliary staff) per diem cost. Resource use within RUG groups was relatively homogeneous. Other predictors of resource use included age, gender and length of stay. RUG-III/22 also met the standard for good reliability (i.e. a kappa value of 0.6 or higher) for crucial classification items, such as activities of daily living and high correlation between assessments based on relative cost. PMID- 10482084 TI - Family planning and family doctors in Estonia. PMID- 10482085 TI - Walking campaigns--a useful way to get people involved in physical activity? Experience from the Stockholm Diabetes Prevention Program (SDPP) PMID- 10482086 TI - Predicting the variation in suicide rates in Norwegian counties. PMID- 10482087 TI - Visual performance after photorefractive keratectomy with a 6-mm ablation zone. AB - PURPOSE: To prospectively examine the effect of photorefractive keratectomy with a 6-mm ablation zone on best-spectacle-corrected visual performance. METHODS: A prospective study was conducted of 164 eyes of 164 patients with an average (+/ SD) of -4.02 +/- 1.74 diopters (range, -0.63 to -8.38 diopters spherical equivalent). Best-spectacle-corrected high-contrast and low-contrast visual acuity (18% Weber contrast) was measured with both natural and dilated pupils. Patients were tested preoperatively and at 3, 6, and 12 months after photorefractive keratectomy. Photorefractive keratectomy was performed with an argon fluoride excimer laser. Fifty-five eyes of 55 patients also underwent astigmatic keratotomy. RESULTS: Twelve months after photorefractive keratectomy, best-spectacle-corrected high-contrast visual acuity with natural pupils showed no significant change from preoperative values; mean (+/-SD) change was 0.004 +/- 0.10 logMAR (t = 0.45, P = .65). Best-spectacle-corrected low-contrast visual acuity with natural pupils was significantly reduced compared to baseline; mean (+/-SD) change was 0.04 +/- 0.13 logMAR (t = 3.3, P = .001). The low-contrast loss was larger (1.5 lines) with dilated pupils; mean (+/-SD) change was 0.13 +/- 0.15 logMAR (t = 9.31, P < .001). Greater losses in dilated low-contrast visual acuity were associated with concurrent astigmatic ketatotomy (t = 2.28, P = .025) and corneal haze of grade 1 or greater (t = 2.71, P = .005). CONCLUSIONS: Reductions in visual performance occur after photorefractive keratectomy with a 6 mm zone. These changes are greatest for low-contrast visual acuity with dilated pupils. Corneal haze and concurrent astigmatic keratotomy are associated with greater losses in low-contrast visual acuity. Best-spectacle-corrected low contrast visual acuity is a sensitive measure for evaluating visual performance after refractive surgery. PMID- 10482088 TI - Acute versus chronic effects of brimonidine on aqueous humor dynamics in ocular hypertensive patients. AB - PURPOSE: To report the acute vs chronic effects of brimonidine, a selective alpha2-adrenergic receptor agonist, on aqueous humor dynamics in ocular hypertensive patients. METHODS: Brimonidine 0.2% was given topically twice daily for 29 days to one eye each of 28 ocular hypertensive volunteers in a randomized double-masked study. The fellow eye was similarly treated with vehicle. Aqueous flow (Fa) and outflow facility (Cfl) were determined with fluorophotometry. Intraocular pressure, outflow facility (Cton), and episcleral venous pressure (Pev) were measured with pneumatonometry, tonography, and venomanometry, respectively. Uveoscleral outflow (Fu) was calculated from intraocular pressure, Fa, Pev, and Cfl values. All measurements were taken on baseline day, day 8, and day 29 of treatment. Intraocular pressure and Fa only were measured after instillation of 1 drop of brimonidine on day 1. RESULTS: When measured 3 hours after instillation on days 1, 8, and 29 of treatment, brimonidine significantly (P < .001) reduced intraocular pressure by at least 5.0 +/- 0.7 mm Hg (mean +/- SEM) compared with baseline day, and by 2.7 +/- 0.5 mm Hg compared with the vehicle-treated contralateral control eyes. The greatest decrease (6.0 +/- 0.6 mm Hg) was observed at 3 hours after the first drop. Aqueous flow was reduced by 29% (P < .001) after the first application but was not significantly different from baseline when measured at day 29 of treatment. Uveoscleral outflow was increased 60% at day 8 (P < .06) and day 29 (P < .05) compared with baseline. There was no significant difference in outflow facility or episcleral venous pressure at day 8 or day 29 of treatment. CONCLUSIONS: The brimonidine-induced reduction in intraocular pressure in humans is associated initially with a decrease in aqueous flow, and after chronic treatment with an increase in uveoscleral outflow. PMID- 10482089 TI - Comparison of 24-hour intraocular pressure reduction with two dosing regimens of latanoprost and timolol maleate in patients with primary open-angle glaucoma. AB - PURPOSE: To compare the 24-hour diurnal ocular hypotensive efficacy of two dosing regimens of latanoprost, once daily (8 AM or 10 PM), vs timolol maleate, twice daily. METHODS: We measured six diurnal intraocular pressure curves (6 AM, 10 AM, 2 PM, 6 PM, 10 PM, and 2 AM) in one randomly selected eye of 34 Greek patients newly diagnosed with primary open-angle glaucoma. The first diurnal curve was an untreated baseline. Patients began taking timolol 0.5%, twice daily, for 2 months. Patients were randomly assigned to latanoprost 0.005% given at 8 AM or 10 PM for 1 month and then changed to the other dosing regimen for 1 month. A diurnal curve was performed after each dosing period. RESULTS: The baseline diurnal pressure for all 34 subjects was 23.1 +/- 3.7 mm Hg. The average intraocular pressures at 6 AM for patients who were given latanoprost in the evening (17.9 +/- 2.9 mm Hg) was statistically lower than that in patients given timolol solution (20.1 +/- 2.5 mm Hg, P = .003); however, patients who were given timolol demonstrated a similar diurnal intraocular pressure (19.1 +/- 2.8 mm Hg) to both morning (18.8 +/- 3.7 mm Hg) and evening doses (18.8 +/- 3.6 mm Hg) of latanoprost (P =.329). When the two latanoprost dosages were compared directly, evening administration provided a statistically lower intraocular pressure at 10 AM (P = .0001) and morning administration at 10 PM (P = .0001). This study had an 80% power to exclude a 1.2-mm Hg difference between groups. CONCLUSIONS: This study indicates that in a small population, both latanoprost and timolol are effective in lowering intraocular pressure throughout a 24-hour period; however, latanoprost is most effective in the 12-hour to 24-hour period after administration. PMID- 10482090 TI - Cataract surgery with ciliary sulcus fixation of intraocular lenses in patients with uveitis. AB - PURPOSE: To describe intentional placement of intraocular lens haptics in the ciliary sulcus of patients with uveitis who are at high risk for postoperative posterior synechiae and lens dislocation. METHODS: We reviewed our experience with 16 eyes of 12 patients with uveitis who underwent cataract surgery with ciliary sulcus fixation of intraocular lenses. Patients were followed for a median of 16.5 months (range, 9 to 44 months) after surgery. We evaluated eyes for surgical technique and the following preoperative and postoperative factors: best-corrected visual acuity, intraocular pressure, anterior chamber cells, and posterior synechiae. The following additional postoperative factors were sought: lens dislocation, lens edge capture, and evidence of pigment dispersion. RESULTS: Posterior synechiae were present in 13 eyes before surgery; postoperative posterior synechiae developed in only three of these eyes. These adhesions resulted in lens edge capture in one eye and limited lens decentration in another. Scant pigment was present on the lens optic or in the anterior chamber, suggesting pigment dispersion, in four eyes. We found no evidence of consistently increased anterior segment inflammation or intraocular pressure after surgery when compared with preoperative levels for this group of patients. Postoperative posterior synechiae were seen more often in eyes that had can-opener anterior capsulotomy than in eyes that had continuous, curvilinear capsulorhexis (P = .036). CONCLUSIONS: Ciliary sulcus fixation allows the intraocular lens to serve as a physical barrier between the iris and the lens capsule remnants. This technique may be useful for reducing the risk of postoperative posterior synechiae in patients with uveitis without increasing the risk of other postoperative problems. PMID- 10482091 TI - Intraocular lens removal from [corrected] patients with uveitis. AB - PURPOSE: To report a series of patients with uveitis and cataract who had undergone cataract extraction with posterior chamber intraocular lens implantation and who subsequently had the intraocular lens removed because of progressive intraocular damage from inflammation. METHODS: Review of the records of 19 patients after removal of a posterior chamber intraocular lens. The decision to perform surgery was based on standard criteria after evaluation at a single uveitis referral center. RESULTS: The complications leading to intraocular lens removal were perilental membrane (eight eyes), chronic low-grade inflammation not responding to anti-inflammatory treatment (eight eyes), and cyclitic membrane resulting in hypotony and maculopathy (three eyes). After intraocular lens removal the inflammation subsided and the visual acuity improved or stabilized in 14 of the 19 eyes. The causes of further reduction in the visual acuity of the other five patients were macular edema (two patients), maculopathy resulting from hypotony (one patient), retinal detachment (one patient), and vitreous hemorrhage (one patient). CONCLUSIONS: Intraocular lens implantation can form part of a reasonable plan for visual rehabilitation of patients with uveitic cataract, but inclusion of an intraocular lens in the plan is not always in the overall long-term best interest of the patient. Intraocular lens removal may salvage useful vision for patients who continue to exhibit complications secondary to uveitis after cataract extraction and intraocular lens implantation, provided the intraocular lens is removed before irreparable damage has been done to macula or optic nerve. PMID- 10482092 TI - The VF-14 index of functional visual impairment in candidates for a corneal graft. AB - PURPOSE: To validate the Visual Function-14 (VF-14) index of functional visual impairment in candidates for a corneal graft. METHODS: One hundred thirty-four patients who were candidates for a corneal graft participated in this study between August 1996 and February 1997. Demographic, ocular history, best corrected visual acuity, and detailed ocular examination data were collected. Functional visual impairment information was obtained by telephone interviews using the following: VF-14, SF-36 (Short Form-36, a more generic measure of general health function), and Visual Symptom Score, and four questions measuring the overall amount of trouble with vision, dissatisfaction with vision, ocular pain, and discomfort. RESULTS: The average age of corneal graft candidates was 64 +/- 18 years (range, 18 to 90 years) and 60% were women. The most frequent corneal disease was pseudophakic bullous keratopathy (41%). Ocular comorbidities included glaucoma or ocular hypertension (30%) and cataract (19%). The mean best corrected visual acuity of the eye scheduled for surgery was 1.33 +/- 0.56 logMAR whereas the best eye best-corrected visual acuity was 0.36 +/- 0.44 logMAR. The mean VF-14 score was 73% +/- 26%, and the internal consistency was high, with a Cronbach alpha value of 0.94. The VF-14 correlated strongly with the best eye best-corrected visual acuity. It also correlated strongly with the Visual Symptom Score, the global measures of trouble and dissatisfaction with vision. Candidates for a corneal graft had low scores for all eight general health concepts evaluated with the SF-36, and the VF-14 correlated with seven of the eight SF-36 subscales. CONCLUSION: The VF-14 is a valid measure of functional visual impairment in candidates for a corneal graft. The Visual Symptom Score and the SF 36 are also useful indices in such patients. PMID- 10482093 TI - Influence of age-related maculopathy on visual functioning and health-related quality of life. AB - PURPOSE: To describe the influence of age-related maculopathy on visual functioning and health-related quality of life. METHODS: A prospective, cross sectional, observational cohort sample of 201 persons with various stages of age related maculopathy was recruited from the Massachusetts Eye and Ear Infirmary as part of a longitudinal study of age-related macular degeneration. Persons were considered to have age-related maculopathy if one or more of the following clinical characteristics were present: drusen, retinal pigment epithelial changes, geographic atrophy, or evidence of exudative disease. Median corrected visual acuity for this sample was 20/25 in the better eye, with all subjects having 20/200 or better visual acuity in at least one eye at baseline. All participants underwent a comprehensive ophthalmologic examination with a dilated pupil. In addition to the usual clinical data collection, severity of age-related maculopathy was graded by an ophthalmologist who used standard clinical criteria and was masked to the participants' descriptions of visual functioning and health related quality of life. All participants completed an interview that included the Activities of Daily Vision Scale, a survey designed to assess difficulties with routine daily activities that require vision, and the Short Form-36 Health Survey, a generic measure of multidimensional health-related quality of life. RESULTS: Severity of age-related maculopathy was associated with poorer scores of the Activities of Daily Vision Scale. This association was most significant for near vision and driving activities. In this sample, the SF-36 Health Survey scales were not significantly correlated with severity of age-related maculopathy. CONCLUSIONS: Reported visual functioning is significantly associated with the clinical severity of age-related maculopathy. However, once visual acuity is taken into consideration, clinical grading of age-related maculopathy did not explain a significant portion of the variation in visual functioning. The lack of significant correlation between severity of age-related maculopathy and the SF-36 Health Survey may have resulted from the small number of participants in our sample with severe bilateral age-related maculopathy. PMID- 10482094 TI - Quality of life of low-vision patients and the impact of low-vision services. AB - PURPOSE: To investigate the functional status and quality of life of patients at a low-vision clinic and to evaluate the impact of low-vision services. METHODS: Interviews, including the Medical Outcomes Study 36-Item Short Form (SF-36), the Visual Function-14 (VF-14), and the 51-item Field Test Version of the National Eye Institute Visual Functioning Questionnaire (NEI-VFQ), were administered to 156 consecutive patients 1 week before and 3 months after their low-vision clinic visit. RESULTS: Low-vision patients scored lower (P < .001) in physical functioning and role limitations caused by physical and emotional health problems than published SF-36 scores of the age-appropriate United States normal population, patients with congestive heart failure, and clinically depressed patients. Low-vision services were associated with improvement in the subjective functional status of 150 patients (98.7%) and were rated "very useful" by 82 (53.9%) patients. The SF-36 scores did not change significantly after low-vision services. The VF-14 mean score improved from 35.8 to 41.2 (P < .001). Four NEI VFQ subscale scores improved significantly (P < .001): general vision, near activities, distance activities, and peripheral vision. CONCLUSIONS: The SF-36, VF-14, and NEI-VFQ demonstrate that low-vision clinic patients perceive marked impairment of functional status and quality of life. Low-vision services are associated with high patient satisfaction. Vision-targeted questionnaires are more sensitive than general health-related quality of life questionnaires to changes in functional status and quality of life after low-vision services, and they may help elucidate the outcomes of low-vision services. PMID- 10482095 TI - Systemic findings associated with central serous chorioretinopathy. AB - PURPOSE: To determine systemic factors associated with central serous chorioretinopathy. METHODS: In a retrospective study, 230 consecutive patients with central serous chorioretinopathy examined in a referral setting were compared with a historical gender-matched and age-matched control group of 230 patients with ocular findings who were examined in the same referral setting. RESULTS: The median age of the patients was 49.8 years, and of the control subjects, 50.0 years. The male-female ratio for both groups was 2.7:1. Patients with central serous chorioretinopathy were more likely to use psychopharmacologic medications (odds ratio = 2.6; 95% confidence interval = 1.30 to 5.19; P = .0049) and corticosteroids (odds ratio = 3.17; 95% confidence interval = 1.30 to 7.70; P = .0067) and were more likely to have hypertension (odds ratio = 2.25; 95% confidence interval = 1.39 to 3.63; P = .0008) than were the control subjects. CONCLUSIONS: This study identified psychopharmacologic medication use, corticosteroid use, and hypertension as factors associated with central serous chorioretinopathy. These findings reinforce the concept that stress and adaptations to stress play a role in this disorder. The findings of possible associations between central serous chorioretinopathy and both hypertension and corticosteroid usage suggest that these modifiable factors may influence morbidity of central serous chorioretinopathy. PMID- 10482096 TI - Retinal vascular occlusion and deficiencies in the protein C pathway. AB - PURPOSE: To report abnormalities in the protein C pathway and other vascular occlusion risk factors in patients with retinal vascular occlusion. METHODS: In a study, we investigated 76 consecutive patients who had in-patient evaluation of venous or arterial retinal vascular occlusion. All patients underwent comprehensive tests for coagulation disorders including determinations of protein C, protein S, lupus anticoagulants, and resistance to activated protein C and were screened for vascular disease risk factors. Resistance to activated protein C was confirmed by a polymerase chain reaction method to detect the specific factor V R506Q mutation. For comparative purposes, we also screened 209 consecutive inpatients with deep vein thrombosis from the same geographic region for resistance to activated protein C as well as protein C and protein S deficiencies. RESULTS: Ten (29%) of 35 patients with central retinal vein occlusion (CRVO) had factor V R506Q mutation. The factor V R506Q mutation was detected in four (19%) of 21 patients with branch retinal vein occlusion. The higher frequency in factor V R506Q mutation compared with the expected 9% mutation prevalence in a white population was highly significant for the central retinal vein occlusion group but not for the branch retinal vein occlusion group. In all patients with resistance to activated protein C, the factor V R506Q mutation was detected; 16 were heterozygous, one homozygous. No cases of lupus anticoagulants, protein C, or protein S deficiencies were detected. Forty (19%) of 209 patients with deep vein thrombosis were carriers of the factor V R506Q mutation. CONCLUSIONS: The prevalence of the factor V R506Q mutation is similar in patients with central retinal vein occlusion and patients with deep vein thrombosis and represents a relevant risk factor. Screening for this mutation is therefore recommended in all patients with central retinal vein occlusion. PMID- 10482097 TI - Color Doppler imaging discloses reduced ocular blood flow velocities in nonexudative age-related macular degeneration. AB - PURPOSE: To study ocular perfusion defects in age-related macular degeneration. METHODS: Twenty-five subjects with nonexudative age-related macular degeneration were compared with 25 age-matched control subjects in studies of flow velocities in several retrobulbar vessels. Color Doppler imaging, which was performed by an examiner who was masked to the subjects' assignment to the control or age-related macular degeneration group, measured peak systolic and end diastolic velocity in the ophthalmic, central retinal, and nasal and temporal posterior ciliary arteries of one eye. A resistive index was calculated from the peak systolic and end diastolic velocity. RESULTS: Subjects with nonexudative age-related macular degeneration showed a consistent trend toward lower peak systolic and end diastolic velocities in the posterior ciliary arteries. For example, in the nasal posterior ciliary artery, the mean end diastolic velocity measured 1.45 +/- 0.34 cm per sec in the age-related macular degeneration group compared with 1.96 +/- 0.66 cm per sec in the control group, yielding a 26% decrease in the age-related macular degeneration group, which represented the largest difference and was highly statistically significant (P = .0012). The resistive index was not significantly altered in the nasal or temporal posterior ciliary artery. Subjects with nonexudative age-related macular degeneration did not differ from control subjects in peak systolic velocity, end diastolic velocity, or resistive index in the ophthalmic artery. In the central retinal artery, the end diastolic velocity was lower (1.37 +/- 1.95 cm per sec vs 1.95 +/- 0.66 cm per sec), whereas the resistive index was higher (0.83 +/- 0.05 vs 0.76 +/- 0.06 cm per sec), in the age-related macular degeneration group; these results were highly statistically significant (P = .0007 and P < .0001, respectively). CONCLUSIONS: Retrobulbar vascular changes in nonexudative age-related macular degeneration subjects include reduced flow velocities in the nasal and temporal posterior ciliary arteries. The reduced peak systolic velocity, combined with the reduced end diastolic velocity at a constant resistive index, seen in nonexudative age related macular degeneration, is consistent with reduced bulk flow in these vessels, suggesting that choroidal perfusion is abnormal in this form of age related macular degeneration. The changes in the central retinal artery suggest there may be a more generalized perfusion abnormality beyond the choroid in patients with age-related macular degeneration or that the central retinal artery exhibits a secondary autoregulatory response to a primary change elsewhere. PMID- 10482098 TI - The influence of ocular pulsatility on scanning laser Doppler flowmetry. AB - PURPOSE: To determine the effect of the cardiac cycle on scanning laser Doppler flowmeter measurements of retinal capillary blood flow in rhesus monkeys and humans. METHODS: Multiple scanning laser Doppler flowmetry images of rhesus monkey and human retinal capillary blood flow over a range of heart rates were obtained. Average flow values were determined for the 64 scan lines that compose the two-dimensional flow map. Cutaneous blood flow was measured simultaneously with a laser Doppler flowmeter. The temporal relationships between retinal capillary blood flow, peripheral arterial pulse, and cutaneous blood flow were determined. In addition, human retinal capillary blood flow in a 10 x 10-pixel area during different phases of the cardiac cycle was compared. RESULTS: Regular oscillations in human and rhesus monkey retinal capillary blood flow are evident as alternating bright and dark horizontal bands in scanning laser Doppler flowmetry images. These fluctuations are temporally correlated with cutaneous blood flow. Linear regression of actual vs predicted heart rate based on peaks in retinal capillary flow yielded r = 0.999 in a rhesus monkey and 0.938 in a human. Retinal capillary blood flow in a 10 x 10-pixel area fluctuated as much as 50% depending on the phase of the cardiac cycle. CONCLUSIONS: The alternating bright and dark banding pattern observed in scanning laser Doppler flowmetry scans of retinal capillary blood flow is related to the cardiac pulse. The errors introduced by pulse-related fluctuations in retinal capillary blood flow are significant and must be minimized or corrected for accurate and reproducible measurements of ocular hemodynamics. PMID- 10482099 TI - Transpupillary thermotherapy for small choroidal melanoma. AB - PURPOSE: To report the treatment of small choroidal melanoma with transpupillary thermotherapy. METHODS: We examined a nonrandomized and uncontrolled series of 14 eyes of 14 patients who were followed up with serial ophthalmoscopy, ultrasonography, and photography. Transpupillary thermotherapy was performed upon documented evidence of tumor growth. RESULTS: After transpupillary thermotherapy, mean follow-up +/- SD was 16 +/- 6.41 months (range, 7 to 28 months) with 10 eyes followed up for at least 1 year. The mean preoperative tumor height was 1.79 +/- 0.59 mm (range, 0.78 to 2.60 mm). Six months after treatment, the mean height was 0.54 mm +/- 0.57 mm (range, 0.00 to 1.16 mm). In 10 eyes, the treated lesion flattened entirely with a mean interval between treatment and flattening of 8.7 months (range, 3 to 21 months). Three patients required retreatment for lack of regression or recurrent growth. The average time to retreatment was 11 months (range, 5 to 15 months). No eye was retreated more than once. There were three amelanotic lesions, all treated in a single session without recurrence. Complications consisted of retinal hemorrhage, retinal vascular occlusion, retinal traction, exudative serous neurosensory detachment, vitreitis, and postoperative pain. The sole treatment failure occurred in an eye treated with a juxtapapillary tumor, with recurrence developing from a previously flattened lesion. This eye was enucleated 10 months after the single initial treatment. At the time of writing, there had been no tumor-related death. CONCLUSIONS: Transpupillary thermotherapy may represent a viable treatment alternative for both pigmented and amelanotic small choroidal melanoma. Diligent follow-up is axiomatic because retreatment may be necessary. Recurrent tumors may develop from flat lesions. Juxtapapillary tumors may be at higher risk for recurrence. Definitive statements regarding the role of transpupillary thermotherapy in the management of small choroidal melanoma await 5-year and 10-year morbidity and mortality data. PMID- 10482100 TI - The emerging role of magnetic resonance angiography in the management of patients with third cranial nerve palsy. AB - PURPOSE: To discuss the sensitivity of three-dimensional time-of-flight (3D TOF) magnetic resonance angiography (MRA) for detecting aneurysms causing third cranial nerve palsy and to propose guidelines for the use of MRA in the evaluation of patients with different clinical presentations of oculomotor nerve palsy. METHODS: From a review of the scientific literature, we determined the proportion of aneurysms of various size that cause third nerve palsy, the sensitivity of MRA in detecting aneurysms of these size classes, and the relationship of aneurysmal size to rupture. RESULTS: The data disclose that properly performed and interpreted 3D TOF MRA will overlook only 1.5% of aneurysms producing third cranial nerve palsy that will, if untreated, rupture during the subsequent 8 years. CONCLUSIONS: Magnetic resonance angiography plays an important diagnostic role in the evaluation of patients with isolated third cranial nerve palsy. However, because of the potentially drastic consequences of overlooking an aneurysm, MRA should be the definitive screening test only among patients with a relatively low likelihood of harboring an aneurysm or a relatively high likelihood of suffering a complication from catheter angiography. PMID- 10482101 TI - Discordance of primary infantile glaucoma in monozygotic twins. AB - PURPOSE: To report a case of primary infantile glaucoma occurring in only one of two identical twins. METHODS: Case reports. RESULTS: Clinical features of bilateral primary infantile glaucoma in one of the two twins at age 16 months are described. Polymorphic DNA markers were used to establish identical twinship. CONCLUSION: Our finding suggests that nongenetic factors must be considered in determining the origin of primary infantile glaucoma. PMID- 10482102 TI - Hyphema associated with pupillary dilation in a patient with exfoliation glaucoma and warfarin therapy. AB - PURPOSE: To describe an unusual hemorrhagic complication associated with pupillary dilation in a patient with exfoliation glaucoma taking anticoagulation therapy. METHODS: A 78-year-old woman with bilateral exfoliation glaucoma who was receiving warfarin, 2 mg daily, for systemic anticoagulation developed acute visual loss in the right eye several hours after pupillary dilation. RESULT: Examination disclosed bilateral advanced exfoliation glaucoma, localized vascularized iridolenticular adhesions in the right eye, and a 4-mm layered hyphema in the right eye. CONCLUSION: Patients with exfoliation glaucoma and vascularized posterior synechiae who are receiving anticoagulation therapy are at increased risk for visually significant spontaneous hyphema after pupillary dilation. PMID- 10482103 TI - Corticosteroid-induced glaucoma attributable to an adrenocorticotropin-secreting malignant carcinoid tumor of the thymus. AB - PURPOSE: To describe the clinical and histopathologic findings in a patient with corticosteroid-induced open-angle glaucoma attributable to an adrenocorticotropin secreting malignant carcinoid of the thymus. METHODS: Case report. In a 33-year old man, the clinical course, laboratory findings, and imaging results as well as the histopathologic findings are described. RESULTS: Increased intraocular pressure in this patient represented a manifestation of severe hypercortisolism attributable to a malignant adrenocorticotropin-secreting carcinoid tumor. Surgical removal resulted in return of the intraocular pressure values to normal levels. CONCLUSION: Thymic carcinoid is a rare cause of Cushing syndrome, which can lead to increased intraocular pressure. PMID- 10482104 TI - Venous collateral remodeling in a patient with posttraumatic glaucoma. AB - PURPOSE: To photographically document venous collateral development, remodeling, and regression in a patient with traumatic glaucoma. METHODS: Consecutive fundus photographs were evaluated, labeled, and correlated with the clinical history of a patient with unilateral posttraumatic glaucoma. RESULTS: This report photographically documents the appearance, remodeling, and subsequent disappearance of collateral vessels from venous occlusion on the surface of the optic disk in an eye with increased intraocular pressure and progressive glaucomatous cupping. CONCLUSIONS: Asymptomatic chronic obstruction of a branch retinal vein on the optic disk may cause venous collaterals to develop in the absence of retinal hemorrhages or other signs of venous occlusive disease. Increased intraocular pressure, arteriolarsclerosis, and glaucomatous cupping are risk factors for these occlusions. PMID- 10482105 TI - Bilateral orbital emphysema from compressed air injury. AB - PURPOSE: To describe a patient who developed bilateral subconjunctival and orbital emphysema after an automobile tire explosion. METHOD: Case report. RESULTS: A 60-year-old man sustained bilateral ocular injury after a tire explosion. Ophthalmic examination disclosed bilateral subconjunctival air, with no visible conjunctival laceration. Computed tomography showed orbital emphysema, with no evidence of orbital fracture. Follow-up examination 2 weeks after the injury disclosed resolution of the subconjunctival air. Best-corrected visual acuity in the right eye was decreased after the explosion but improved to the baseline level of 20/40 2 weeks after the injury. CONCLUSION: Subconjunctival and orbital emphysema can occur from high-pressure air injury in the absence of an obvious entry site. PMID- 10482107 TI - Optical cross-sectional assessment of the macula by retinal thickness analyzer in optic disk pit maculopathy. AB - PURPOSE: To assess the morphologic changes in optic disk pit maculopathy. METHOD: Case report. A 45-year-old woman with optic disk pit maculopathy in the left eye was examined with the scanning retinal thickness analyzer before and after vitrectomy with intravitreal gas tamponade. RESULTS: Preoperatively, optical cross-sectional retinal thickness analyzer images clearly demonstrated the extent of retinoschisis and outer layer detachment in the macula. Postoperatively, the retinal thickness analyzer images showed that retinal thickening decreased as the intraretinal fluid in the schisis cavity was absorbed and then gradually decreased further as the outer layer detachment in the fovea resolved. CONCLUSIONS: Scanning retinal thickness analyzer can be used to diagnose and monitor the structural changes of the macula lesion in optic disk pit maculopathy. Vitrectomy with gas tamponade may be effective for treating this disease. PMID- 10482106 TI - Leu518Pro mutation of the beta ig-h3 gene causes lattice corneal dystrophy type I. AB - PURPOSE: To describe a Japanese family with lattice corneal dystrophy type I, which segregates with a novel mutation, Leu518Pro of the beta ig-h3 gene. METHODS: DNA was extracted from leukocytes in four members (three affected and one unaffected) of a Japanese family with lattice corneal dystrophy type I. Exon 12 of the beta ig-h3 gene was amplified and analyzed with a molecular biologic method. Clinical data were also collected. RESULTS: Three generations of this family have been positively diagnosed with lattice corneal dystrophy, indicating autosomal dominant inheritance. We found a heterozygous point mutation that segregates with the disease phenotype. It was a single base-pair transition (CTG to CCG, Leu to Pro). CONCLUSION: Although it is extremely rare compared with the Arg124Cys mutation of the beta ig-h3 gene, Leu518Pro mutation of the beta ig-h3 also causes lattice corneal dystrophy type I. PMID- 10482108 TI - Hypotensive ischemic optic neuropathy and peritoneal dialysis. AB - PURPOSE: To report anterior ischemic optic neuropathy associated with systemic hypotension in a patient undergoing continuous ambulatory peritoneal dialysis. METHODS: Case report. A 58-year-old man undergoing continuous ambulatory peritoneal dialysis developed painless blurred vision in both eyes and bilateral optic disk swelling with an altitudinal field defect in the left eye. Twenty-four hour ambulatory blood pressure monitoring was requested in addition to other routine investigations. RESULTS: Routine blood pressure measurement in the clinic was 130/86 mm Hg, but ambulatory blood pressure monitoring demonstrated pronounced early morning hypotension with individual readings as low as 91/41 mm Hg. CONCLUSIONS: Renal dialysis can render patients hypotensive, and this may be associated with anterior ischemic optic neuropathy. The overnight drop in blood pressure may not be appreciated with routine blood pressure measurement. Therefore, 24-hour ambulatory blood pressure monitoring should be considered when investigating patients with suspected anterior ischemic optic neuropathy who are undergoing renal replacement. PMID- 10482109 TI - Topless optic disk syndrome without maternal diabetes mellitus. AB - PURPOSE: To describe four cases of topless optic disk syndrome without maternal diabetes mellitus. METHOD: Four patients had incidentally discovered inferior visual field defects. RESULTS: Ophthalmoscopic examinations in all four patients disclosed superiorly displaced entrances of the central retinal artery and thinning of the superior peripapillary nerve fiber layers. One patient had a superior peripapillary crescent with pallor of the superior disk. These clinical findings were consistent with a diagnosis of superior segmental optic hypoplasia, the topless disk. None of the patients had mothers who had diabetes. CONCLUSIONS: The topless optic disk syndrome can occur in the absence of maternal diabetes mellitus. PMID- 10482110 TI - Mitochondrial maculopathy: geographic atrophy of the macula in the MELAS associated A to G 3243 mitochondrial DNA point mutation. AB - PURPOSE: To report ocular findings in the mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS syndrome) in a family with the A to G 3243 mitochondrial (mt) DNA point mutation. METHODS: Case reports. Ocular findings are described from four family members with the MELAS associated A to G 3243 mt DNA point mutation. RESULTS: Findings included ophthalmoplegia, neurosensory deafness, reduction of photopic and scotopic electroretinogram b wave amplitudes, and myopathy, as well as macular retinal pigment epithelial atrophy. No family members had nyctalopia, attenuation of retinal blood vessels, or retinal bone spicule pigmentation. CONCLUSION: The finding of slowly progressive macular retinal pigment epithelial atrophy expands the reported phenotypic diversity of patients with A3243G mt DNA mutations. PMID- 10482111 TI - Castleman disease presenting with ophthalmic signs and symptoms. AB - PURPOSE: To describe a patient with multicentric Castleman disease who was initially examined with ophthalmic signs and symptoms. METHODS: Case report. A 71 year-old man was initially examined with swelling of both upper eyelids and diplopia of 2 months' duration. RESULTS: Medical evaluation and right axillary lymph node biopsy disclosed Castleman disease. Systemic corticosteroid treatment temporarily resolved signs and symptoms, but the patient died of recurrence with cytomegalovirus and Aspergillus infection 10 months after initial examination. CONCLUSIONS: Multicentric Castleman disease is a rare but distinct disorder that may present initially with ocular signs and symptoms. This disease must be included in the differential diagnosis of orbital pseudotumor and lymphoma. PMID- 10482112 TI - Control of visual symptoms in two men with birdshot retinochoroidopathy using low dose oral corticosteroid therapy. AB - PURPOSE: To describe two men with birdshot retinochoroidopathy whose severe subjective visual symptoms were controlled by long-term use of low-dose oral corticosteroid therapy. METHOD: Chart review. RESULTS: Vision improved subjectively in both men after initial treatment with oral prednisone (1 mg/kg/day) and remained stable while taking 5 mg daily or less of prednisone for periods of 54 and 81 months. CONCLUSION: Some patients with birdshot retinochoroidopathy do well with no long-term therapy or only low-dose oral corticosteroids as long-term therapy. PMID- 10482113 TI - Adie syndrome: evidence for refractive error and accomodative asymmetry as the cause of amblyopia. AB - PURPOSE: To report objective changes in accommodation in a child with Adie syndrome. METHODS: A child aged 2 years 10 months when initially examined was found to have good visual acuity in both eyes, a low degree of hypermetropia (isometropic), and Adie pupil presumed to be caused by chicken pox that had occurred 2 months earlier. Amblyopia developed but responded well to treatment, which involved correction of the refractive error and occlusion therapy. Objective changes in the refraction of the eye were measured on the Canon R1 autorefractor at 3.8 m and 33 cm. RESULT: The degree of accommodation in the affected eye when both eyes were open was markedly reduced. CONCLUSION: The presence of isometropic hypermetropia, which remains uncorrected when Adie syndrome is present, can lead to the development of amblyopia in a child. PMID- 10482114 TI - Spontaneous regression of orbital Langerhans cell granulomatosis in a three-year old girl. AB - PURPOSE: To report a case of spontaneous regression of orbital Langerhans cell granulomatosis. METHOD: Case report. A 3-year-old girl was initially examined with a 5-week history of slowly progressive blepharoptosis and periorbital swelling of the left eye. RESULTS: Computed tomographic scan showed a mass in the left orbit eroding into the left frontal bone; fine-needle aspiration confirmed diagnosis of Langerhans cell granulomatosis. After initial biopsy, the patient was treated by close observation alone. Six months after initial examination, the monostotic lesion had completely resolved. CONCLUSIONS: In some cases of monostotic Langerhans cell granulomatosis, initial biopsy followed by observation alone may allow for the spontaneous regression of the lesion. This conservative approach to treatment is an important therapeutic option that may spare the patient the adverse effects of surgical resection, radiation, or chemotherapy. PMID- 10482115 TI - Orbital floor implant migration across the ethmoidal sinuses and nasal septum. PMID- 10482116 TI - Ultrasound biomicroscopic quantitative analysis of light-dark changes associated with pupillary block. PMID- 10482117 TI - Thrombin-antithrombin III complex in acute retinal vein occlusion. PMID- 10482118 TI - A brief history of psychiatry: millennia past and present--part III. AB - Release of emotion through a variety of means has often been seen as healing. Throughout the centuries many cultures dealt with stress by bringing relief through methods like dancing, ritual, or confession. The idea of discovering not always conscious emotions and their release through talking became important in 19th-century psychological treatments. The 20th century has seen a flowering of psychoanalytical, behavioral, cognitive behavioral, and many other psychotherapies. These have been used in individual, group, or martial/family settings, inpatient and outpatient. There have been disagreements between and among the various schools of psychotherapy. As we approach the next century, therapists of all schools are discussing ways in which each therapy contributes to helping patients and are defining common threads that link all the approaches to psychotherapy. PMID- 10482119 TI - Outcome of psychogeriatric intervention in an old-age home: a 3 years follow-up study. AB - Psychogeriatrics and especially psychiatric services in nursing and old-age homes are sorely lacking in research. Four years ago we developed and implemented a model for service delivery for old-age homes within our hospitals' catchment area. In the first year of operation 48 of 373 residents (13%) were evaluated and treated by a psychogeriatrician. The aim of the present study was to evaluate outcome of these patients three years later. All patients and medical records were re-assessed. During the three-year period 16/48 subjects (33.3%) passed away; of these half were previously diagnosed as suffering from a major depressive episode. None of the subjects who died had attempted suicide. Following a preliminary diagnosis of affective spectrum disorder, 4/48 patients (8.3%) were re-diagnosed at follow up as suffering from dementia. Full or partial remission of the original symptomatology was achieved in 18/32 surviving subjects (50.7%); mostly depressed or anxious patients. We conclude that although mortality rate among elderly psychiatric patients is high persistent interventions lead to successful remission in the majority of patients. PMID- 10482120 TI - Experience with the atypical antipsychotics--risperidone and olanzapine in the elderly. AB - There is paucity of published data regarding controlled trials with risperidone and olanzapine in elderly psychotic patients. Medical records of 151 hospitalized geropsychiatric patients (risperidone patients n = 114 and olanzapine patients n = 37) were analyzed for demographic data, target symptoms, doses, effects, side effects, comorbid medical conditions and concurrent medications. The mean age of the patients was 71 years. The male: female ratio was essentially the same for both groups. The mean daily dose was 3 mg for risperidone and 10 mg for olanzapine. 78% of the risperidone group and 75% of the olanzapine group appear to have responded to treatment. The discontinuation rates of medication was the same in both groups (22%). Adverse events were reported in 16-17% in both groups. It appears from this study that both risperidone and olanzapine are relatively safe and effective in geropsychiatric patients with comorbid medical illnesses. Controlled studies and head-to-head comparison studies are recommended. PMID- 10482121 TI - Diagnoses, psychosocial stressors and adaptive functioning in attempted suicide. AB - A systematic sample of 78 suicide attempters (37 men and 41 women), of whom 83% were hospitalized, were interviewed according to SCID I and II and Axes III-V according to DSM-III-R. Mood disorders were most common (56%). Forty-four suicide attempters (56%) suffered from comorbid diagnoses on Axis I-II. Borderline personality disorder was more common among women then men (56% vs. 24%, respectively, p = 0.01). Axis III disorders were confirmed for 45%. Sixty-two percent of the suicide attempters had severe psychosocial stressors (Axis IV). When comparing subjects with only Axis I disorders to those with Axis I and II disorders, no difference with respect to psychosocial stressor grade was observed. Moreover, those with only Axis I disorders were not impaired in their adaptive functioning (Axis V) even if severe psychosocial stressors were present. In contrast, an association (p = 0.02) was found between high stress and low functioning in patients with both Axis I and Axis II disorders. The data suggest that in clinical practice, beside evaluation of Axis I and Axis II disorders, also stressors and global functioning should be included in the assessment of suicide risk after attempted suicide. PMID- 10482122 TI - Cigarette smoking and psychiatric disorder in a community sample. AB - We examined the lifetime prevalence of psychiatric disorders in cigarette smokers and nonsmokers in a non-patient sample. First-degree relatives of psychiatric patients (n = 697) and normal controls (n = 360) were interviewed with the Diagnostic Interview Schedule and the Structured Interview for DSM-III Personality Disorders. Using these interviews we diagnosed the major mental (Axis I) disorders and personality (Axis II) disorders. A cigarette smoker was defined as someone who smoked daily for a month or more at some time in their lives. We found that smokers more frequently had a lifetime history of major depression, alcohol and drug abuse/dependence, agoraphobia, unstable/acting out and anxious/fearful personality disorders. In a logistic regression analysis, the only significant variables independently associated with smoking status were the alcohol and drug use disorders. Age was an important modifying variable--the smoking-illness relationship was robust in the youngest age cohort and negligible in the oldest cohort. We conclude that cigarette smokers have increased rates of mood, anxiety, substance use, and personality disorders. However, after controlling for the comorbidity among the disorders only alcohol and drug abuse/dependence were independently associated with smoking. Young smokers had particularly high rates of substance use disorders. This age effect may reflect the impact of a quarter century of health education. PMID- 10482123 TI - The use of quetiapine for treatment-resistant bipolar disorder: a case series. AB - OBJECTIVE: To determine if quetiapine, an atypical antipsychotic agent approved for the treatment of schizophrenia, is effective in the treatment of bipolar disorder. MATERIALS AND METHODS: A retrospective chart review identified six patients with DSM-IV bipolar disorder, type I, who received open uncontrolled treatment with quetiapine in the setting of nonresponse or intolerance to traditional mood-stabilizing treatments. Treatment response was based on moderate to marked improvement on the Clinical Global Impression-Bipolar Disorder (CGI-BP) scale. RESULTS: Two of six patients showed evidence of response. The main side effect noted was sedation. CONCLUSIONS: Quetiapine may be a useful treatment for some patients with treatment-resistant bipolar disorder. Further studies are needed to assess quetiapine's effect more rigorously. PMID- 10482124 TI - Quetiapine as an alternative to clozapine in the treatment of dopamimetic psychosis in patients with Parkinson's disease. AB - There are many difficulties associated with the late stages of Parkinson's disease (PD), but psychosis and agitation may be the most disturbing for both patients and care givers, and often precipitate the pivotal decision for long term nursing home placement. While the addition of antipsychotic drugs or the withdrawal of antiparkinsonian drugs may improve the behavioral problem, these strategies usually worsen the motor difficulties. Clozapine has been studied in PD for over a decade, and while it appears to be effective, there are safety and tolerability concerns associated with it. In addition, in New Jersey, Medicaid no longer pays for the home blood draws that are required for home-bound patients. This led to a situation in which we had patients who needed to stop clozapine and begin an alternative therapy. Because quetiapine seems particularly well suited to patients with PD based on in vitro and in vivo studies we have begun to try this medication in PD patients who need to stop clozapine. This article reports three case histories of patients with PD, confusion and dopamimetic psychosis who had been previously managed with clozapine and who were successfully switched to quetiapine. At doses from 12.5 to 150 mg/day quetiapine was well tolerated, resulting in behavioral improvement and no real increase in parkinsonism. These case histories raise the possibility that quetiapine may represent a viable alternative to clozapine in PD patients with dopamimetic psychosis and behavioral disturbances. PMID- 10482125 TI - The metabolism of atypical antipsychotic drugs: an update. AB - This paper reviews the current literature describing the metabolism of both multi receptor clozapine analogue atypical antipsychotic drugs (clozapine, olanzapine, and quetiapine) and serotonin-dopamine antagonist atypical antipsychotic drugs (risperidone, sertindole and ziprasidone), to highlight the significance of those data in the context of clinical practice. The former group of atypical antipsychotic drugs shares a similar tricyclic structural nucleus and are metabolized through three major categorical metabolic pathways--N+-oxidation, N glucuronidation, and phases 1 and 2 biotransformation with final glucuronidation before renal excretion. Differing in clozapine and olanzapine, quetiapine has incomplete data describing its metabolism. The latter group of atypical antipsychotic drugs has diversified chemical structures and absence of data on N+ oxidation and N-glucuronidation in the literature. But their metabolic routes in phase 1 biotransformation are versatile although current data are far from completion. No apparent significant drug interactions in clinical practice are reported, although QT prolongation is implicated in all those three drugs. None of all six atypical antipsychotic drugs are identified as significant inhibitors or inducers to any co-administered medication. The author suggests the need for more research to address some pertinent clinical issues in the metabolism of those drugs. PMID- 10482126 TI - Impediments to the discharge planning effort for psychiatric inpatients. AB - OBJECTIVE: This study investigates a methodology to systematically track the effort to overcome impediments to securing needed post-hospital care and support. METHODS: 494 consecutive admissions to the Mount Sinai Medical Center were evaluated for the quality of available support resources in the domains of housing, daily activity, and psychiatric treatment using the Mount Sinai Discharge Planning Inventory. On day seven, the same rater recorded the optimal discharge plan for each resource category and those impediment codes that were likely to interfere with attaining the desired resource. On discharge, the rater recorded the actual resources obtained for the patient, and if the plan did not meet the patient's needs, an impediment code was used to explain why the sub optimal "discharge fit" was not secured. RESULTS: Having an impediment in any of the three resource categories (housing, daily activities, psychiatric treatment services) at day 7 was predictive of a sub-optimal discharge plan. Of all three resource categories studied, a decline in overall impediments from day 7 to discharge was significant only for psychiatric treatment services. An internal/clinical impediment in any of the three resource categories on day 7 was associated with a patient history of alcohol and drug abuse. A significant association was found between having external/environmental impediments identified at discharge for housing and psychiatric treatment services with return to the hospital within 90 days of discharge. CONCLUSION: Study of the impediments to the discharge planning effort provides an opportunity to elucidate the factors that comprise the pathway of recovery from psychiatric illness, but which are normally ill-defined, poorly understood, or not readily measured. PMID- 10482127 TI - Women's help-seeking patterns for depression. AB - Depression tends not to be accurately identified and treated in primary care settings. Women of color are especially likely to use these settings for mental health issues but are less likely to be diagnosed appropriately. A study was conducted within four Florida primary care clinics serving primarily low-income families. Participants included 321 women (Black, 22%, Hispanic, 23.5%, White, 38.6%) who completed a confidential questionnaire while waiting to be seen by clinic staff. Ten percent reported recent major depression, with 26.7% indicating depressive symptoms during the past two years. Depressed women were significantly more likely to report physical violence during the past year. Respondents turned primarily to family, friends, and medical clinics for their depression. They found turning to clinic staff to be helpful and described psychosocial interventions as useful. Members of all ethnic/racial groups reported barriers to seeking help, including perceived separation between mental health and general health and stigma. Implications are discussed in terms of appropriate community education and screening procedures. PMID- 10482128 TI - The role of social work in HIV/AIDS clinical trials. AB - The social worker can facilitate screening, retention and patient adherence in HIV/AIDS clinical trials. This paper introduces the process and its key vocabulary, and uses three case studies to demonstrate how social workers can assist clients who may wish to participate in, or are already enrolled in a clinical trial. After examining five major issues that affect the client in a clinical trial (informed consent; treatment vs. research; risks and side effects; altruism; the role of family members; and gender, race and class issues), the authors elaborate on interventions at the screening level, and concrete services and psychosocial interventions for study participants. PMID- 10482129 TI - The role of the supervisor in training social workers treating HIV infected persons in a hospital. AB - The role of social workers working with HIV infected persons in hospitals is complex and laden with sources of professional stress, including those related to the hospital framework and environment and personal stress related to patients, their families, and the workers themselves. The paper identifies these factors of stress and discusses the functions of the supervision in avoiding or reducing them. The article is based on experience in Israel, but the analysis may be applicable elsewhere, as well. PMID- 10482130 TI - Unraveling the mysteries of high density lipoprotein metabolism. PMID- 10482131 TI - Hepatocellular transport and secretion of biliary lipids. AB - Bile is the route for elimination of cholesterol from the body. Recent studies have begun to elucidate hepatocellular, molecular and physical-chemical mechanisms whereby bile salts stimulate biliary secretion of cholesterol together with phospholipids, which are enriched (up to 95%) in phosphatidylcholines. Active translocation of bile salts and phosphatidylcholines across the hepatocyte's canalicular plasma membrane provides the driving force for biliary lipid secretion. This facilitates physical-chemical interactions between detergent-like bile salt molecules and the ectoplasmic leaflet of the canalicular membrane, which result in biliary secretion of cholesterol and phosphatidylcholines as vesicles. Within the hepatocyte, separate molecular pathways function to resupply bile salts, phosphatidylcholines and cholesterol to the canalicular membrane for ongoing biliary lipid secretion. PMID- 10482132 TI - Contribution of cholesterol 7alpha-hydroxylase to the regulation of lipoprotein metabolism. AB - Clinical studies have clearly established a relationship between bile acid synthesis and plasma LDL-cholesterol concentrations. Interruption of the enterohepatic circulation of bile acids leads to increased bile acid synthesis and a reduction in plasma LDL-cholesterol concentrations. New studies indicate that genetic variation in cholesterol 7alpha-hydroxylase activity accounts for a significant fraction of the inter-individual variation in plasma LDL-cholesterol concentrations in the general population, and a specific CYP7A1 allele associated with increased plasma LDL-cholesterol concentrations has been identified. Studies in which cholesterol 7alpha-hydroxylase was transiently overexpressed in hamsters and mice indicate that direct manipulation of cholesterol 7alpha-hydroxylase leads to changes in plasma LDL-cholesterol concentrations. Interestingly, targeted inactivation of the gene encoding cholesterol 7alpha-hydroxylase does not lead to increased plasma LDL-cholesterol concentrations in mice. PMID- 10482133 TI - Metabolism of high density lipoprotein subfractions. AB - Over the past few years, new experimental approaches have reinforced the awareness among investigators that the heterogeneity of HDL particles indicates significant differences in production and catabolism of HDL particles. Recent kinetic studies have suggested that small HDL, containing two apolipoprotein A-I molecules per particle, are converted in a unidirectional manner to medium HDL or large HDL, containing three or four apolipoprotein A-I molecules per particle, respectively. Conversion appears to occur in close physical proximity with cells and not while HDL particles circulate in plasma. The medium and large HDL are terminal particles in HDL metabolism with large HDL, and perhaps medium HDL, being catabolized primarily by the liver. These novel kinetic studies of HDL subfraction metabolism are compelling in-vivo data that are consistent with the proposed role of HDL in reverse cholesterol transport. PMID- 10482134 TI - Intestinal cholesterol absorption. AB - The strong association between intestinal cholesterol absorption and total plasma cholesterol level has renewed interest in the absorptive process and stimulated the generation of new animal models. Increasingly, new studies suggest that cholesterol absorption is genetically controlled and supports a protein-mediated mechanism for cholesterol uptake into the intestinal mucosal cell. Insights into potential mechanisms are predicted to lead to novel pharmacological approaches to inhibit cholesterol absorption. PMID- 10482135 TI - Cholesteryl ester transfer protein and its plasma regulator: lipid transfer inhibitor protein. AB - The interconnections between cholesteryl ester transfer protein (CETP) expression and lipid metabolism, and the possible roles of CETP in atherogenesis are examined. The importance of lipid transfer inhibitor protein in modulating CETP activity is detailed, and the consequences of this inhibitory activity on CETP mediated events are proposed. PMID- 10482138 TI - Bibliography. Current world literature. Lipid metabolism. PMID- 10482136 TI - Scavenger receptor BI and cholesterol trafficking. AB - Scavenger receptor BI (SR-BI) mediates the selective uptake of HDL cholesteryl ester into steroidogenic cells and the liver and is a major determinant of the plasma HDL concentration in the mouse. Recent studies indicate that SR-BI also alters the metabolism of apolipoprotein B-containing particles and influences the development of atherosclerosis in several animal models. These results and the similar pattern of SR-BI expression in humans emphasize that it is important to learn how this receptor influences lipoprotein metabolism and atherosclerosis in people. PMID- 10482137 TI - The assembly and secretion of apolipoprotein B-containing lipoproteins. AB - The assembly of lipoproteins containing apolipoprotein B is a complex process that occurs in the lumen of the secretory pathway. The process consists of two relatively well-identified steps. In the first step, two VLDL precursors are formed simultaneously and independently: an apolipoprotein B-containing VLDL precursor (a partially lipidated apolipoprotein B) and a VLDL-sized lipid droplet that lacks apolipoprotein B. In the second step, these two precursors fuse to form a mature VLDL particle. The apolipoprotein B-containing VLDL precursor is formed during the translation and concomitant translocation of the protein to the lumen of the endoplasmic reticulum. The VLDL precursor is completed shortly after the protein is fully synthesized. The process is dependent on the microsomal triglyceride transfer protein (MTP). Although the mechanism by which the lipid droplets are formed is unknown, recent observations indicate that the process is dependent on MTP. The fusion of the two precursors is not dependent on MTP, but the mechanism remains to be elucidated. The conversion of the apolipoprotein B containing precursor to VLDL seems to be dependent on the ADP ribosylation factor 1 (ARF 1) and its activation of phospholipase D. During their assembly, nascent apolipoprotein B chains undergo quality control and are sorted to degradation. Such sorting, which occurs cotranslationally during the formation of the apolipoprotein B-containing precursor, involves cytosolic chaperons and ubiquitination that targets apolipoprotein B to proteasomal degradation. Other levels of sorting occur in the secretory pathway. Thus, lysosomal enzymes are involved as well as the LDL receptor. PMID- 10482139 TI - Genetics and molecular biology. PMID- 10482140 TI - Lipid metabolism. PMID- 10482141 TI - Hyperlipidaemia and cardiovascular disease. PMID- 10482142 TI - Atherosclerosis: cell biology and lipoproteins. PMID- 10482143 TI - Therapy and clinical trials. PMID- 10482144 TI - Comparison of immediate and follow-up results of the short and long NIR stent with the Palmaz-Schatz stent. AB - The intrinsic characteristics of a stent including stent length may affect both procedural success and long-term outcome. The present study evaluated the immediate and follow-up results after implantation of the short and long NIR stent and compared these results with the Palmaz-Schatz stent. Between July 1995 and December 1996, stenting with a 16-mm NIR stent (NIR-16), a 32-mm NIR stent (NIR-32), or a Palmaz-Schatz stent (PS) was performed in 68, 57, and 155 lesions, respectively. There were no significant differences in the incidences of delivery failure (PS, 2.6%: NIR-16, 4.4%; NIR-32, 5.3%; p = NS) and procedural success (PS, 92%; NIR-16, 93%; NIR-32, 93%; p = NS) among the 3 groups. The reference vessel diameter was smaller in lesions with a 32-mm NIR stent than in those with a Palmaz-Schatz stent (PS, 3.14+/-0.58, NIR-16, 3.00+/-0.50; NIR-32, 2.90+/-0.47 mm; p <0.05). The lesion length was longer in lesions with a 32-mm NIR stent than in those with a Palmaz-Schatz or a 16-mm NIR stent (PS, 8.9+/-5.0; NIR-16, 11.0+/ 4.1; NIR-32, 26.1+/-9.7 mm; p <0.01). After the procedure, the lesions with a 32 mm NIR stent had a smaller minimal lumen diameter than those with a Palmaz-Schatz stent (PS, 3.17+/-0.61; NIR-16, 2.99+/-0.51; NIR-32, 2.89+/-0.49 mm; p <0.01). At follow-up, a smaller minimal lumen diameter was observed in lesions with a 32-mm NIR stent than in those with a Palmaz-Schatz or a 16-mm NIR stent (PS, 2.32+/ 0.98; NIR-16, 2.25+/-0.80; NIR-32, 1.68+/-0.79 mm; p <0.01). Restenosis rates were 16.5% in lesions with a Palmaz-Schatz stent, 13.3% in those with a 16-mm NIR stent, and 47.4% in those with a 32-mm NIR stent (p <0.01). Although stent delivery and procedural success of a long NIR stent were acceptable, the restenosis rate of a long NIR stent was high compared with a short NIR stent or a Palmaz-Schatz stent. PMID- 10482145 TI - Primary coronary infarct artery stenting in acute myocardial infarction. AB - Completed and ongoing randomized trials have provided results that favor primary infarct-related artery (IRA) stenting as opposed to primary percutaneous transluminal coronary angioplasty, but the applicability of the trial results to all patients with acute myocardial infarction (AMI) has not yet been investigated. This study sought to determine the applicability of an unconditional IRA stenting strategy in nonselected patients with AMI. After successful mechanical recanalization of the IRA, all patients with AMI and a reference diameter > or =2.5 mm were considered eligible for primary IRA stenting without any restriction regarding age or clinical status on presentation. The primary end point of the study was a composite end point defined as death, reinfarction, or repeat target lesion revascularization. Primary IRA stenting was successfully performed in 161 of 190 consecutive patients with AMI (85%), and of 162 (99%) considered suitable for stenting. Patients with nonstented IRA had a reference IRA diameter smaller than patients with a stent (2.71+/-0.48 vs 3.20+/ 0.41 mm, p <0.001). Overall, the 6-month mortality was 5%. Mortality was 2% for patients without, and 32% for patients with cardiogenic shock. The incidences of reinfarction and of repeat target lesion revascularization were 1% and 12%, respectively. The 6-month angiographic follow-up showed an IRA patency rate of 94% and a restenosis rate of 26%. The results of this study strengthen the hypothesis that unconditional primary IRA stenting is highly feasible, and may actually improve the outcome of patients with AMI. PMID- 10482146 TI - Comparison of cilostazol versus ticlopidine therapy after stent implantation. AB - The aim of this study was to evaluate the efficacy of cilostazol for prevention of stent thrombosis compared with ticlopidine. Cilostazol is a potent antiplatelet agent with less serious side effects. However, few data are available about the effect of cilostazol in preventing stent thrombosis after coronary stent implantation. Four hundred ninety patients selected for elective stent placement were randomized to receive aspirin plus ticlopidine (n = 243) or aspirin plus cilostazol (n = 247) for 1 month. Clinical and laboratory evaluations were performed at regular interval. There were no differences in baseline characteristics between the 2 groups. During the first 30 days after stent implantation, major cardiac events or adverse drug effects were similar between the 2 groups: ticlopidine (2.9%) vs cilostazol (1.6%) group, p = NS; stent thrombosis (0.4% vs 0.8%, p = NS, respectively), myocardial infarction (0.4% vs 0.8%, p = NS), severe leukopenia (1.2% vs 0%, p = NS), severe thrombocytopenia (0.4% vs 0%, p = NS), and cerebral hemorrhage (0.4% vs 0%, p = NS). Adverse effects led to drug withdrawal in 7 patients in the ticlopidine group (2.9%) and in 5 in the cilostazol group (2.0%). There was no death during the follow-up period. Thus, aspirin plus cilostazol may be an effective antithrombotic regimen with comparable results to aspirin plus ticlopidine after elective coronary stenting. PMID- 10482147 TI - Frequency and prognostic value of cardiac troponin I elevation after coronary stenting. AB - Mild myocardial injuries after coronary angioplasty are associated with adverse late outcomes. The incidence and prognostic value of this phenomenon when using cardiac troponin I (cTnI) after stent implantation is unknown. We studied cTnI and creatine kinase (CK) release in 109 patients after stenting. Clinical success was achieved in 103 patients (94%). In-hospital major adverse coronary events were: death in 1 patient, Q-wave myocardial infarction in 1 patient, and non-Q wave myocardial infarction in 2 patients. Twenty-nine patients (27%) had postprocedural cTnI increase, 16 (15%) had CK elevation. No preprocedural variables predicted marker elevation. Marker release was related to the occurrence of in-lab complications (59% vs 29% [p = 0.004 for cTnI] and 69% vs 32% [p = 0.011 for CK]). In 34% no explanation was found for cTnI increase. Success was more frequent in patients without cTnI elevation (100% vs 86%, p <0.001). The negative predictive value of cTnI increase was 100% for in-hospital major adverse coronary events (MACE), whereas its positive predictive value was 14%. cTnI and CK concordant elevation was associated with more intra- and postprocedural adverse events. During a mean follow-up of 8+/-3 months, major adverse coronary events were: death in 2 patients, myocardial infarction in 2 patients, and repeat PTCA in 8 patients. cTnI elevation was not predictive of these late MACE. cTnI elevation is common after stenting, and is related to the occurrence of in-lab complications. Its isolated elevation is not a good predictor of MACE. Patients with concordant cTnI and CK elevation seem to be at higher risk of in-hospital MACE. PMID- 10482148 TI - Platelet activation as a potential mechanism of GP IIb/IIIa inhibitor-induced thrombocytopenia. AB - The blockade of the platelet integrin glycoprotein (GP) IIb/IIIa has proved to be an effective antiplatelet therapy. Profound thrombocytopenia has repeatedly been described as an adverse effect in patients treated with GP IIb/IIIa inhibitors, but its mechanism has not been elucidated yet. With use of flow cytometry, the activation status of platelets was monitored in 26 patients presenting with acute myocardial infarction who were treated with the GP IIb/IIIa inhibitor abciximab alone or in combination with the fibrinolytic agent reteplase. Fibrinogen and PAC 1 (a GP IIb/IIIa activation-specific monoclonal antibody) binding, as well as P selectin expression on unstimulated platelets were constant in 25 patients throughout a follow-up of 7 days. In 1 patient (D.F.), the percentage of platelet binding fibrinogen increased from 2.2% to 17.8%, for PAC-1 from 2.8% to 13.2%, and for P-selectin expression from 10.2% to 58.3% 10 minutes after the start of treatment. Furthermore, D.F. had a decrease in single platelet count in ethylenediaminetetraacetic acid-, citrate-, and heparin-anticoagulated and native blood. Blood films revealed platelet aggregates. In vitro testing of D.F.'s blood 2 and 4 weeks after initial admission demonstrated a reinduction of fibrinogen and PAC-1 binding to platelets, an increase of P-selectin expression, and formation of platelet aggregates following exposition of platelets to abciximab in vitro. In summary, this report describes the induction of platelet activation by a GP IIb/IIIa inhibitor in vivo and reinduction in vitro in direct association with thrombocytopenia. Platelet activation by GP IIb/IIIa inhibitors may be one potential mechanism for GP IIb/IIIa inhibitor-induced thrombocytopenia. PMID- 10482149 TI - Baroreflex and non-baroreflex modulation of vagal cardiac control after myocardial infarction. AB - Vagal control of sinus node exerted by arterial baroreceptors is markedly impaired 48 hours after acute myocardial infarction (AMI), but it recovers 10 days later. However, it is unknown whether this recovery is peculiar to baroreceptor vagal control or reflects normalization of the overall vagal modulation of heart rate. In 21 untreated patients (aged 51+/-3 years, mean +/- SEM) studied 10+/-1 and 21+/-1 days after an AMI and in 13 healthy controls (aged 47+/-2 years), we examined the increases in RR interval (electrocardiogram) induced by carotid baroreceptor stimulation via a neck chamber and by immersion of the face in iced water for 15 seconds (diving reflex). Both 10 and 21 days after AMI, baseline blood pressure and RR interval values were superimposable to those obtained in controls. Ten days after AMI, the bradycardic responses to carotid baroreceptor stimulation were similar to those seen in controls (maximal RR interval lengthenings: 248+/-34 vs 270+/-31 ms, respectively, p = NS) and remained virtually unchanged later. In contrast, the bradycardic response to diving was reduced in patients after AMI compared with controls (maximal RR interval lengthenings: 203+/-43 vs 325+/-52 ms, respectively, p <0.05) and did not improve later. Thus, in AMI recovery of the early impairment of baroreceptor heart rate control does not reflect normalization of vagal cardiac control, which remains lower than normal values at a time when the baroreflex is restored. PMID- 10482150 TI - Terminal QRS distortion on admission is better than ST-segment measurements in predicting final infarct size and assessing the Potential effect of thrombolytic therapy in anterior wall acute myocardial infarction. AB - We assessed predicting final infarct size (using predischarge Selvester score) by 3 electrocardiographic variables in 267 patients with first anterior wall acute myocardial infarction (AMI) undergoing (n = 86) or not undergoing (n = 181) thrombolysis. Patients with previous AMI or inverted T waves in leads with ST elevation were excluded. The sum (sigma) of ST elevation, the number of leads with ST elevation, and the initial electrocardiographic pattern were determined on the admission electrocardiogram (absence (QRS-) or presence (QRS+) of distortion of the terminal portion of the QRS in > or =2 leads (J point > or =0.5 of the R-wave amplitude in leads I, aVL, V4 to V6, or presence of ST elevation without S waves in leads V1 to V3). There was no association between sigmaST elevation and final infarct size in patients who did or did not receive thrombolytic therapy. Analysis of covariance showed that the number of leads with ST elevation (F = 19.6), thrombolysis (F = 25.2), and QRS+ initial pattern (F = 19.5) were all associated with final infarct size (p <0.0001 for all). Among patients who did not receive thrombolytic therapy, the average Selvester score was 19.7+/-9.9 for the QRS- patients and 26.1+/-10.4 for the QRS+ patients (p = 0.02). Among patients who received thrombolytic therapy, the average Selvester score was 11.7+/-9.8 for the QRS- patients and 24.2+/-10.1 for the QRS+ patients (p <0.0001). Thrombolysis reduced final Selvester score only in the QRS- group (p <0.00001), but not in the QRS+ group (p = 0.45). It is concluded that (1) final Selvester score in anterior wall AMI can be predicted by the number of leads with ST elevation, the initial electrocardiographic pattern, and thrombolysis, and (2) thrombolysis reduces final Selvester score only in patients with QRS- pattern. PMID- 10482151 TI - Significance of T-wave changes during early dobutamine stress echocardiography in patients with Q-wave acute myocardial infarction. AB - The relation between T-wave changes and regional contraction during dobutamine stress echocardiography at low (5 to 10 microg/kg/min) and high (20 to 40 microg/kg/min) doses in 43 consecutive patients, early (7+/-2 days) after first recent Q-wave acute myocardial infarction has been evaluated. T-wave changes detected in > or =2 infarct-related electrocardiographic leads during dobutamine infusion were defined as follow: (1) negative T waves becoming positive, (2) positive T waves becoming upright > or =2 mm, and (3) negative T waves becoming upright > or =2 mm from baseline. Wall motion score index (WMSI) was defined as the sum of the echocardiographic scores of 16 segments divided by total segments considered at baseline, and at low and peak doses of dobutamine. Patients were classified according to the absence or presence of dobutamine T-wave changes. Those without T-wave changes had a significantly higher WMSI at rest (1.68+/-0.23 vs 1.50+/-0.21; p <0.05) and at peak (1.77+/-0.34 vs 1.51+/-.30 p <0.05) of dobutamine stress testing, without higher incidence of viability, homozonal, and heterozonal ischemia and chest pain. The angiographic patterns were similar between groups. Regression analysis showed a significant correlation between WMSI and T-wave amplitude at baseline (R = 0.38, p = 0.01) and at peak dobutamine stress testing (R = 0.50, p = 0.0006). The sensitivity sensitivity, specificity, and accuracy of T-wave changes to detect myocardial viability were 0.27, 0.84, and 0.70, respectively. The sensitivity, specificity, and accuracy of T-wave changes to detect homozonal ischemia were 0.76, 0.27, and 0.46, respectively. In conclusion, dobutamine-induced T-wave changes are associated with a greater extent of wall motion abnormalities both at rest and at peak stress echocardiography, but they are of little value in predicting myocardial viability when analyzed early after myocardial infarction. PMID- 10482152 TI - Association of lipids and lipoprotein level with total mortality and mortality caused by cardiovascular and cancer diseases (Poland and United States collaborative study on cardiovascular epidemiology). AB - This study evaluates the relation between total serum cholesterol, triglycerides, and high-density lipoprotein (HDL) cholesterol, and subsequent total, cardiovascular, and cancer mortality. These data are from 4,946 US and 5,198 Polish men and women aged 35 to 64 years at baseline with mortality follow-up over 13 years. Total cholesterol showed a U-shaped or J-shaped relation to age adjusted total and cancer mortality across all samples, with significance only in Polish women. The multivariable adjusted relative risk for total and cancer mortality was higher in the lowest cholesterol category only in Poland and significant only for cancer. Cardiovascular mortality was positively related to cholesterol, but only in Polish men and US women was mortality significantly higher in the highest versus the lowest cholesterol category. The multivariable adjusted relative risk of cardiovascular death was greater in the highest versus the lowest cholesterol category, but this trend was significant only in the US. HDL cholesterol was inversely related to total (significant only in US men) and cardiovascular mortality (significant only in US and Polish men). A similar, but not significant, association of HDL cholesterol was found with cancer mortality. The multivariable adjusted relative risk of total mortality was inversely related to HDL cholesterol significant in both the US and Poland. The relative risk of cardiovascular mortality was significantly lower at higher HDL cholesterol levels in all samples. The relative risk of cancer mortality was highest and significant at the lowest HDL cholesterol level in the US and Poland. Elevated triglycerides were associated with increased risk of total and cardiovascular mortality, but this trend was significant only in the US. Cancer mortality was not significantly related to triglycerides. The present study indicates that in geographically and culturally diverse populations, the relation of lipids with cardiovascular mortality is similar. The relation with total and cancer mortality varies by country, gender, and lipids. This suggests that relations of total and cancer mortality with lipids or lipoproteins are weaker than associations with cardiovascular mortality. PMID- 10482153 TI - Effect of exogenous estrogen on atherothrombotic vascular disease risk related to the presence or absence of the factor V Leiden mutation (resistance to activated protein C). AB - Estrogen replacement therapy (ERT), which produces acquired resistance to activated protein C when superimposed on heritable resistance to activated protein C (the mutant Factor V Leiden trait), may promote venous and arterial thrombosis. In a cross-sectional study of 423 women referred for hyperlipidemic therapy (93 of whom [22%] were on ERT), our specific aim was to determine whether ERT and heterozygosity for the Factor V Leiden mutation and/or resistance to activated protein C interacted as risk factors for atherothrombosis. Of the 423 women, 168 (40%) had atherothrombosis, 19 (4%) were heterozygous for Factor V Leiden mutation or had resistance to activated protein C <2 (Factor V Leiden mutation+), and 404 were wild-type normal for the Factor V gene and/or had resistance to activated protein C > or =2 (Factor V Leiden mutation-). By stepwise logistic regression, positive explanatory variables for atherothrombosis included hypertension (p = 0.002), age (p = 0.003), relatives with atherothrombosis (p = 0.002), anticardiolipin antibody immunoglobulin-M (p = 0.02), and a Factor V Leiden mutation*ERT interaction term where atherothrombosis events were more likely in 2 subgroups of women (ERT- and Factor V Leiden mutation-) or (ERT+ and Factor V Leiden mutation+) (p = 0.02). High-density lipoprotein cholesterol was inversely associated with atherothrombosis (p = 0.004). In a separate logistic regression model for the 213 women with a polymerase chain reaction measurement of the Factor V gene, ERT was protective (p = 0.008); the Factor V Leiden mutation was positively associated with atherothrombosis (p = 0.05). The atherothrombosis odds ratio risk for ERT (yes vs no) was 0.36 (95% confidence intervals [CI] 0.16 to 0.74, p = 0.007). The atherothrombosis risk odds ratio in women heterozygous for the Factor V Leiden mutation (vs normal) was 2.00 (95% CI 1.02 to 4.22, p = 0.05). ERT may be protective against atherothrombosis when the Factor V Leiden mutation is absent, whereas the Factor V Leiden mutation may increase risk for atherothrombosis, particularly in the presence of ERT. We suggest that the Factor V Leiden mutation be measured in all women on ERT or before beginning ERT to identify those heterozygous for the Factor V Leiden mutation (4%), in whom ERT is relatively or absolutely contraindicated because of increased risk for atherothrombosis and thromboembolism. A second, much larger group of women will also be identified without the factor V Leiden mutation (96%), in whom ERT may reduce the risk for atherothrombosis. PMID- 10482154 TI - Interaction of septadian and circadian rhythms in life-threatening ventricular arrhythmias in patients with implantable cardioverter-defibrillators. AB - Previous studies have shown that life-threatening ventricular arrhythmias display both circadian and septadian (day of the week) periodicity. We hypothesized that assessing the relation between these circadian and septadian rhythms may provide important pathophysiologic information about the mechanism of sudden cardiac death. Using the database from a population of 683 consecutive patients with a third-generation implantable cardioverter-defibrillator (ICD), we examined the time pattern of ICD activations for rapid (prospectively defined as cycle length <280 ms) tachycardias for each day of the week. A total of 5,270 arrhythmic episodes were analyzed. Despite the fact that event distribution was significantly nonuniform (p <0.001) for both circadian and septadian analyses, the circadian pattern was strikingly similar for each day of the week with a relatively broad peak between 9 A.M. and 6 P.M. and a long nadir between 9 P.M. and 6 A.M. We conclude that the trigger factors responsible for the daily circadian distribution of life-threatening ventricular arrhythmias in a population with ICDs are similar throughout the week and may thus be unrelated to the standard work week. These data suggest that the physiologic modulators of circadian and septadian rhythms may be different. PMID- 10482155 TI - Relations of diastolic left ventricular filling to systolic chamber and myocardial contractility in hypertensive patients with left ventricular hypertrophy (The PRESERVE Study). AB - Abnormalities of left ventricular (LV) diastolic filling and stress-corrected midwall shortening (MWS) have been described in hypertensive patients with normal ejection fraction (EF). However, whether stress-corrected MWS parallels LV diastolic filling better than EF does remains uncertain. Blood pressure, body mass index, echocardiographic LV mass and LV geometry, EF and stress-corrected MWS, LV diastolic filling (peak E- and A-wave velocities, E-wave deceleration time, and atrial filling fraction) were evaluated in 212 hypertensive patients with LV hypertrophy enrolled in the Prospective Randomized Enalapril Study Evaluating Regression of Ventricular Enlargement study. LV structure, geometry, as well as LV diastolic filling, were compared between patients with reduced EF (<55%, n = 39, 18%) and those with normal EF (>55%) as well as between patients with reduced stress-corrected MWS (<89.2%, n = 31, 15%) and those with normal stress-corrected MWS (>89.2%). Patients with reduced EF had higher LV mass, eccentric LV geometry, and higher heart rate than those with normal EF, although they did not differ in age, blood pressure, or body mass index. LV filling pattern was also similar in those 2 groups. Patients with reduced stress corrected MWS had higher atrial filling fraction, body mass index, heart rate, LV mass, and concentric geometry than those with normal stress-corrected MWS. Atrial filling fraction was negatively associated with stress-corrected MWS, but not with EF in multivariate models, independently of age, gender, heart rate, and body mass index. Thus, in hypertensive patients with LV hypertrophy, abnormal LV diastolic filling is more closely related to impaired myocardial contractility than to LV chamber EF. PMID- 10482156 TI - Comparison of frequencies of left ventricular systolic and diastolic heart failure in Chinese living in Hong Kong. AB - There is a wide variation (13% to 74%) in the reported prevalence of heart failure associated with normal left ventricular (LV) systolic function (diastolic heart failure). There is no published information on this condition in China. To ascertain the prevalence of diastolic heart failure in this community, 200 consecutive patients with the typical features of congestive heart failure were studied with standard 2-dimensional Doppler echocardiography. A LV ejection fraction (LVEF) >45% was considered normal. The results showed that 12.5% had significant valvular heart disease. Of the remaining 175 patients, 132 had a LVEF >45% (75%). Therefore, 66% of patients with a clinical diagnosis of heart failure had a normal LVEF. Heart failure with normal LV systolic function was more common than systolic heart failure in those >70 years old (65% vs 47%; p = 0.015). Most (57%) had an abnormal relaxation pattern in diastole and 14% had a restrictive filling pattern. In the systolic heart failure group, a restrictive filling pattern was more common (46%). There were no significant differences in the sex distribution, etiology, or prevalence of LV hypertrophy between these 2 heart failure groups. In conclusion, heart failure with a normal LVEF or diastolic heart failure is more common than systolic heart failure in Chinese patients with the symptoms of heart failure. This may be related to older age at presentation and the high prevalence of hypertension in this community. PMID- 10482157 TI - Cardiac positron emission tomography imaging with [11C]hydroxyephedrine, a specific tracer for sympathetic nerve endings, and its functional correlates in congestive heart failure. AB - The integrative mechanisms of autonomic dysfunction in congestive heart failure (CHF) remain poorly understood. We sought to study cardiac retention of [11C]hydroxyephedrine (HED), a specific tracer for sympathetic presynaptic innervation, and its functional correlates in CHF. Thirty patients with mild to moderate heart failure underwent resting cardiac HED positron emission tomography imaging, spectrum analysis testing of systolic pressure and heart rate variability in the resting supine and 70 degrees head-up tilt positions, and testing of baroreflex sensitivity. Compared with control subjects, global myocardial HED retention index was reduced by 30% (p <0.01) in patients with CHF. The HED retention index did not correlate significantly with heart rate variability. However, it correlated with baroreflex sensitivity at rest (r = 0.43, p = 0.05) and with systolic pressure low-frequency (0.03 to 0.15 Hz) variability at head-up tilt (r = 0.76, p <0.01), as well as with low-frequency systolic pressure variability response from baseline to tilt (r = 0.75, p <0.01). We conclude that cardiac HED retention is reduced in patients with CHF. This correlates with blunted vascular sympathetic effector responses during posture induced reflex activation and baroreflex control of heart rate, suggesting an interdependence between cardiac presynaptic innervation abnormalities and neural mechanisms important to blood pressure maintenance in CHF. PMID- 10482158 TI - Comparison of long-term results of percutaneous mitral balloon valvotomy with closed transventricular mitral commissurotomy at a single North American Institution. AB - Long-term (>3 years) follow-up data were obtained from 102 consecutive patients undergoing percutaneous mitral balloon valvotomy (PMBV). Data were collected prospectively by review of the medical record, mailed questionnaire, and/or telephone. Data on patients with closed mitral commissurotomy (CMC) at our institution have been previously reported and serve as the comparison group. Follow-up data was 98% complete at a mean of 57 months for PMBV patients. Compared with patients undergoing CMC, these patients were older (54+/-14 vs 43.6+/-10 years, p <0.001) and more likely to have undergone previous mitral valve surgery (17% vs 4%, p <0.001). The observed 5-year survival in the PMBV group was no different from that observed in the CMC group (83% vs 90%, p = NS) or from that predicted by the model developed from the CMC patients. Commissural calcium was associated with death and death or repeat mitral valve procedure in the multivariate analysis. Long-term survival free from repeat procedures was equivalent when patients with commissural calcium were excluded. Thus, PMBV offers long-term survival and freedom from subsequent mitral valve procedures similar to CMC. PMID- 10482159 TI - Usefulness of low-dose dobutamine stress echocardiography for evaluating reversibility of brain death-induced myocardial dysfunction. AB - Many of the myocardial wall motion abnormalities in heart donors are reversible after transplantation, indicating that the presence of wall motion abnormalities should not automatically lead to the exclusion of donor hearts. The present study observes the natural course of brain death-induced myocardial dysfunction, and investigates whether low-dose dobutamine stress echocardiography could identify reversible myocardial dysfunction in brain-dead patients. We prospectively measured the serial changes of left ventricular fractional shortening (FS) using echocardiography and cardiac troponin T from admission to the time of cardiac standstill in 30 brain-dead patients. Patients were divided into 2 groups according to FS at the time of brain death; group I (FS > or =30%) and group II (FS <30%). Dobutamine stress echocardiography was performed in group II. Twenty three patients were in group I and 7 patients were in group II. Four patients among 7 patients in group II showed dobutamine-nonresponsive wall motion (group IIa) and the remaining 3 patients showed dobutamine-responsive wall motion (group IIb). Troponin T at the time of brain death was markedly higher in group IIa than in groups I and IIb (5.13+/-3.79 vs 0.23+/-0.20, 0.22+/-0.16 ng/ml, p <0.0001, respectively). FS remained normal and troponin T was not increased until cardiac standstill in group I. FS remained decreased and troponin T remained elevated until cardiac standstill in group IIa, whereas FS became normal at 7 days after brain death with no change in troponin T in group IIb. Thus, some brain death induced myocardial dysfunction is reversible and low-dose dobutamine stress echocardiography may identify reversible myocardial dysfunction. PMID- 10482161 TI - Angiographic findings in patients undergoing catheterization for recurrent symptoms within 30 days of successful coronary intervention. AB - In 108 consecutive patients without abrupt vessel closure referred for repeat coronary angiography within 30 days of successful coronary intervention, 28 (26%) were found with restenosis at the treated site. None of the 27 patients who underwent stenting were found to have early restenosis; balloon angioplasty without stenting was the only independent predictor of early restenosis in patients with recurrent symptoms within 30 days of intervention. PMID- 10482160 TI - Patterns of body fat deposition in youth and their relation to left ventricular markers of adverse cardiovascular prognosis. AB - The patterns of body fat deposition in healthy youth and their relation to future development of cardiovascular disease remain incompletely understood. To further evaluate these patterns, we measured indirect indexes of central and general fat deposition in healthy adolescents (mean age 15.4+/-2.3 years) with family histories of hypertension. We examined the relation between these indexes and echocardiographic markers of adverse prognosis as well as the effect of gender and ethnicity. All 225 subjects (64% black and 48% female) had > or =1 biologic parent and 1 grandparent with hypertension. Skinfold thicknesses, waist-to-hip girth ratio, Quetelet index, Ponderal index, conicity, and Z score weight - Z score height were measured. Left ventricular (LV) mass, indexed LV mass, relative wall thickness (RWT), and midwall fractional shortening (MFS) were determined using echocardiography. In both black and white subjects, the adiposity indexes were significantly correlated with posterior wall thickness, total LV mass, and indexed LV mass (p <0.05 for all). Additionally, in black subjects, central adiposity was inversely related to MFS and directly related to RWT and septal thickness. General adiposity independently predicted indexed and nonindexed LV mass, whereas central adiposity predicted MFS and RWT. Compared with subjects with normal LV geometry, those with abnormal geometry were heavier and fatter based on every index of obesity (p <0.03 for all). Thus, indexes of fat deposition are significantly correlated with LV markers of adverse prognosis in healthy youth. PMID- 10482162 TI - Clinical characteristics and outcome in postinfarction pseudoaneurysm. AB - The clinical characteristics and outcome in 22 patients with postinfarction pseudoaneurysm were studied. The medium-term outcome was poor (median follow-up 3.6 years), but cardiac rupture was uncommon in patients treated conservatively. PMID- 10482163 TI - Lack of association of restenosis following coronary angioplasty with elevated C reactive protein levels or seropositivity to Chlamydia pneumoniae. AB - Seventy-five consecutive patients undergoing directional coronary atherectomy were evaluated by measuring anti-Chlamydia immunoglobulin G and anticytomegalovirus immunoglobulin G antibodies, and serum levels of C-reactive proteins (before atherectomy). The results showed that although both Chlamydia infection and elevated C-reactive protein levels are associated with coronary artery disease and coronary artery disease events, neither of these appears to play a role in the development of restenosis. PMID- 10482164 TI - Direct myocardial revascularization and angiogenesis--how many patients might be eligible? AB - This study examines the proportion of patients with ischemic heart disease who may be candidates for the newer modalities of revascularization. A significant proportion (approximately 5%) of patients who undergo coronary angiography at tertiary referral centers may be eligible for newer methods of therapy. PMID- 10482165 TI - Early results of percutaneous intervention for severe coexisting carotid and coronary artery disease. AB - Fifty-one patients with severe coexisting carotid and symptomatic coronary artery occlusive disease successfully underwent staged or simultaneous coronary angioplasty and carotid stenting. One pericardial effusion and 2 minor strokes with full recovery occurred in the hospital, but no major neurologic events, myocardial infarction, or death were observed and no repeat revascularization was required within the 30-day follow-up. PMID- 10482167 TI - Experience with a single-pass, dual-electrode implantable atrial defibrillator lead for maintaining sinus rhythm in patients with recurrent atrial fibrillation. AB - The implantable atrial defibrillator is a new potential nonpharmacologic treatment for recurrent atrial fibrillation. The results of this study suggest that a simplified lead configuration, with a single-pass, dual-electrode atrial defibrillation lead can be used for both atrial fibrillation detection and defibrillation with an implantable atrial defibrillator. PMID- 10482166 TI - Relation between low-density lipoprotein cholesterol and thoracic aortic atherosclerosis. AB - This prospective study, which included 320 patients, showed that total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, total cholesterol/high-density lipoprotein cholesterol, and triglycerides correlate with thoracic aortic atherosclerosis. Low-density lipoprotein cholesterol is identified as an independent predictor of thoracic aortic plaque related to the severity of thoracic aortic atherosclerosis. PMID- 10482168 TI - Clinical significance of elevated levels of cardiac troponin T in patients with chronic heart failure. AB - Cardiac troponin T, measured by second-generation assay, is a highly sensitive and specific marker of minor myocardial damage. Cardiac troponin T was elevated in patients with severe chronic heart failure; it identifies those with latent and progressive myocardial damage and those who are at increased risk of cardiac events. PMID- 10482169 TI - Comparison of incidences of congestive heart failure in older African-Americans, Hispanics, and whites. AB - In a prospective study of 2,893 African-Americans, Hispanics, and whites, mean age 81 years, at 43-month follow-up, congestive heart failure (CHF) developed in 194 of 686 African-Americans (28%), in 67 of 257 Hispanics (26%), and in 533 of 1,950 whites (27%) (p = NS). The Cox regression model showed that significant independent risk factors for CHF were male gender (risk ratio = 1.4, p = 0.0001); hypertension (risk ratio = 2.5, p = 0.0001); coronary artery disease (risk ratio = 4.0, p = 0.0001); diabetes mellitus (risk ratio = 1.6, p = 0.0001); and age (risk ratio = 1.05, p = 0.0001). PMID- 10482170 TI - Inferior vena caval masses identified by echocardiography. AB - The most common cause of an inferior vena caval mass is renal cell carcinoma that extends through the lumen, occurring in 47 of 62 patients (85%). Detection of an inferior vena caval mass affects the surgical approach requiring cardiopulmonary bypass for resection when the mass extends to the heart. PMID- 10482171 TI - Significance of a Q wave in lead I in the newborn. AB - We present 11 healthy newborns whose electrocardiograms had a pure Q wave in lead I and who did not have a myocardial infarction clinically. We propose that in the healthy newborn, a pure Q wave in lead I may be due to increased right ventricular mass, not myocardial infarction. PMID- 10482172 TI - Prospective diagnosis of alveolar capillary dysplasia in infants with congenital heart disease. AB - Alveolar capillary dysplasia (ACD) is a lethal pulmonary disorder found in newborns that is characterized by severe pulmonary hypertension and hypoxemia. We report on the clinical behavior of this disorder in a series of patients and its association with congenital heart disease, especially left heart obstructive disease; we also report a prospective diagnosis of ACD by lung biopsy in a newborn with congenital heart disease, which prevented futile and prolonged medical intervention. PMID- 10482173 TI - A symposium: sodium-hydrogen exchange inhibition. Introduction. PMID- 10482174 TI - Protection of the myocardial cell during ischemia. AB - Prevention of myocardial necrosis in acute coronary syndromes is the immediate goal of therapy. Decreasing myocardial oxygen needs is of marginal value. Improving oxygen delivery by mechanical or thrombolytic reperfusion is more successful but still leaves much to be desired in terms of time to reperfusion before damage occurs due to reperfusion itself. During ischemia, there is a metabolic mismatch between glycolysis and glucose oxidation that results in accumulation of hydrogen ions, which, in turn, activates the Na+/H+ exchange system (NHE-1), leading to Na+ and Ca2+ overload and cell death. Blocking NHE-1 is a new strategy designed to prevent or delay cell death. Cariporide, a potent inhibitor of the NHE-1 system, is currently under investigation. Other agents under investigation are designed to modify proton generation, modify proton effects, and attenuate necrosis progression. Also under study are agents designed to mediate preconditioning (adenosine agonists and adenosine triphosphate sensitive potassium channel openers). Other approaches to minimize cell injury include use of antioxidants and free-radical scavengers, complement inhibitors, selectin blockers, and glycoprotein (GP) IIb/IIIa antagonists. PMID- 10482175 TI - Rational basis for use of sodium-hydrogen exchange inhibitors in myocardial ischemia. AB - The cardiac sarcolemmal Na+/H+ exchanger extrudes intracellular H+ in exchange for Na+, in an electroneutral process. Of the 6 mammalian exchanger isoforms identified to date, the Na+/H+ exchanger (NHE)-1 is believed to be the molecular homolog of the sarcolemmal Na+/H+ exchanger. The exchanger is activated primarily by a reduction in intracellular pH (intracellular acidosis), although such activation is subject to modulation by a variety of endogenous mediators (e.g., catecholamines, thrombin, endothelin) through receptor-mediated mechanisms. A large body of preclinical evidence now suggests that inhibition of the sarcolemmal Na+/H+ exchanger attenuates many of the unfavorable consequences of acute myocardial ischemia and reperfusion. Much of this evidence has been obtained with recently developed potent, selective inhibitors of the exchanger, such as HOE-642 (cariporide) and its structurally related congener HOE-694, in studies using both in vitro and in vivo models of ischemia and reperfusion in a variety of species. The data from these studies indicate that Na+/H+ exchange inhibition leads to a decreased susceptibility to severe ventricular arrhythmia, attenuates contractile dysfunction, and limits tissue necrosis (i.e., decreases infarct size) during myocardial ischemia and reperfusion. Such protection is likely to arise, at least in part, from attenuation of "Ca2+ overload," which has been linked causally with all of these pothologic phenomena. The consistent and marked cardioprotective benefit that has been observed with cariporide and related compounds in preclinical studies suggests that Na+/H+ exchange inhibition may represent a novel and effective approach to the treatment of acute myocardial ischemia in humans. PMID- 10482176 TI - Sodium-hydrogen exchange inhibition: novel strategy to prevent myocardial injury following ischemia and reperfusion. AB - Activation of Na+/H+ exchange and subsequent calcium overload in cardiac myocytes appear to play an important role in myocardial tissue injury following ischemia and reperfusion. Results of several in vitro studies in isolated myocytes and heart preparations and in vivo studies in pigs and rats have suggested that inhibition of Na+/H+ exchange is an effective means to prevent lethal reperfusion injury, arrhythmia, and improve myocardial contractile dysfunction. In patients with acute myocardial infarction (MI), any preventive agent is administered immediately before or shortly after reperfusion, rather than before the occurrence of coronary occlusion. The direct interventional approach to treating acute MI provides the opportunity to see if reperfusion has already occurred; if not, a protective agent prior to mechanical reperfusion by percutaneous transluminal coronary angioplasty (PTCA) can be administered to limit reperfusion injury. In a multicenter, randomized, placebo-controlled clinical trial, we tested the hypothesis that inhibition of Na+/H+ exchange with cariporide (HOE 642) could limit infarct size and improve myocardial function in patients with acute transmural MI treated with direct PTCA. Patients were randomized to receive placebo or cariporide given as a 40-mg intravenous bolus prior to reperfusion. Global and regional left ventricular function were analyzed via paired contrast left ventriculograms performed before direct PTCA and after 21 days. Myocardial enzymes (i.e., creatine kinase [CK], CK-MB, and lactate dehydrogenase) as markers for myocardial tissue injury were evaluated as well. The results of this pilot study suggested that the Na+/H+ exchange inhibition could be of benefit to prevent reperfusion injury in patients with acute anterior MI treated with direct angioplasty. PMID- 10482178 TI - Have the recently completed phase III trials significantly impacted adjuvant treatment choices for stage II breast carcinoma? PMID- 10482177 TI - GUARD During Ischemia Against Necrosis (GUARDIAN) trial in acute coronary syndromes. AB - The plasma membrane sodium-hydrogen exchange system plays a major role in the pathophysiology of myocardial ischemia and reperfusion injury. The GUARD During Ischemia Against Necrosis (GUARDIAN) trial was undertaken to assess the potential clinical benefits of cariporide, a novel inhibitor of the sodium-hydrogen exchanger, in patients with acute coronary syndromes at risk of myocardial necrosis. This prospective, doubleblind, randomized, multicenter trial enrolled 11,733 patients who had unstable angina/non-Q-wave myocardial infarction (MI), or who required high-risk percutaneous interventions or coronary bypass surgery. Strict entry criteria were applied to ensure the enrollment of a population at high risk of developing complications. The patients were randomized to receive intravenous cariporide 20, 80, or 120 mg every 8 hours or placebo every 8 hours. Treatment was applied for the period of risk between 48 hours and 7 days. The primary efficacy endpoint was a composite of all-cause mortality and MI at 36 days. Secondary endpoints included the occurrence of the composite endpoint at day 10, events related to left ventricular dysfunction at day 36 and 6 months, extent of infarction, and refractory ischemia at day 36. Enrollment was completed in August 1998, and the results were presented at the American College of Cardiology meeting in New Orleans, Louisiana, USA, in March 1999. PMID- 10482179 TI - Introduction: a therapeutically relevant framework for the classification of human prostate cancer. PMID- 10482180 TI - African American men, prostate cancer early detection examination use, and informed decision-making. AB - It is well known that African American men are more likely to be diagnosed with metastatic prostate cancer than White men. Racial variation in the use of prostate cancer early detection modalities (ie, digital rectal examination [DRE] and prostate-specific antigen [PSA] testing) has been suggested as a major reason for this differential. Several factors may help to explain the reported low levels of DRE and PSA test utilization among African American men, including background sociodemographic characteristics, medical history, and cognitive and psychosocial perceptions. In this review, the impact of these characteristics on prostate cancer early detection examination utilization is explored. Findings from studies showing race-related differences in cognitive and psychosocial factors are presented. Preparatory education for informed decision-making is suggested as an approach to help minimize racial differences in cognitive and psychosocial factors that influence the use of prostate cancer early detection modalities. The need to facilitate informed decision-making along the continuum of care is highlighted. PMID- 10482181 TI - Implications of cell death regulation in the pathogenesis and treatment of prostate cancer. AB - Although the phenomenon of programed cell death, or apoptosis, has been recognized for many years, interest in the clinical implications of apoptotic cell death has not been widely apparent until recently. It is now established that the products of at least some oncogenes and tumor suppressor genes are able to regulate the rates of apoptosis as well as proliferation in tumor cell populations. In fact, it appears that evasion of deletion via apoptotic cell death may be a requisite event in the development of many malignant neoplasms. Molecular alterations that occur at high frequency in the pathogenesis of prostate cancer often involve genes implicated in the regulation of cell death. There is now ample reason to speculate that the susceptibility of individual malignant neoplasms to undergo apoptosis in response to a given therapeutic intervention may be a useful parameter in predicting therapeutic response. This realization has profound implications with respect to the design and implementation of treatment strategies for prostate cancer based on the biology of this disease. PMID- 10482182 TI - Markers of androgen-independent progression of prostatic carcinoma. AB - Prostate cancer (PCa) remains the most common cancer and the second leading cause of cancer mortality in men in the United States. The evolution from a localized to a metastatic phenotype coupled with the progression from an androgen-dependent (AD) to an androgen-independent (AI) state leads to a universally fatal disease. Identifying the biologic characteristics associated with PCa progression is a major goal of current research efforts by different groups, in the hope to better predict the natural history of the disease in an individual patient and to design treatments based on the specific biologic behavior. PMID- 10482183 TI - The role of the androgen receptor in the development and progression of prostate cancer. AB - Androgens are essential for the development, growth, and maintenance of the prostate. They exert their effects via the intracellular androgen receptor (AR), which is a ligand-dependent transcription activator. As is the case with normal prostate development, primary prostatic cancers are largely dependent on androgens for growth and survival. Most patients respond favorably to androgen ablation and antiandrogen therapy, which has become a standard treatment of metastatic disease. However, virtually all patients will relapse with clinically defined androgen-independent cancer. This phenomenon raises the question of how cancer cells survive and grow in the low androgen environment? Two of the routes cells can take to adapt are (1) bypassing and (2) sensitizing the AR pathway. The vast numbers of AR abnormalities observed in prostate tumors from patients treated with hormonal therapy suggest that many cells sensitize or change the AR pathway. To continue to activate this pathway in a low androgen environment, cells can (1) mutate the AR to become promiscuously activated by different steroids, (2) amplify the AR, (3) activate the AR in a ligand-independent manner by growth factors and cytokines, or (4) amplify coactivators. Alternatively, prostate cancer cells that have lost AR expression must have bypassed the AR pathway. Activation of oncogenes and autocrine growth factor stimulation are two mechanisms that likely contribute to becoming completely androgen-independent. From all the studies on AR function in prostate cancer, it is clear that the AR plays an important role in cancer development and progression. Moreover, the AR pathway remains important in most cells from patients with clinically defined androgen-independent prostate cancer. PMID- 10482184 TI - Metastasis-related genes in prostate cancer. AB - The identification of genes specifically related to the development of metastatic disease in prostate cancer is complicated by tumor cell heterogeneity and the presence of expressed sequences that are not related to metastasis. A system was designed to minimize these complications using differential display-polymerase chain reaction (DD-PCR) together with genetically paired cell lines derived from primary mouse prostate cancer and their associated metastases generated in vivo by the metastatic mouse prostate cancer reconstitution model. Using this system, a number of metastasis-related sequences were identified, including a cDNA that encodes caveolin-1. PMID- 10482185 TI - Chemotherapy in advanced prostate cancer. AB - In the United States, prostate cancer is the most commonly diagnosed cancer and the second leading cause of cancer death in men. Prostate cancer is a rare disease before age 40; however, the prevalence increases quickly to 80% by the age of 80, and with increasing life expectancy, hormone-refractory prostate cancer (HRPC) will soon represent the most common cancer in the male population in the United States and other Western countries. The evolution of early prostate cancer is variable and extends over many years; some tumors progress slowly or not at all, whereas others may progress more rapidly and be fatal after a few years. A widely held view is that chemotherapy has no role in HRPC because no single agent or combination has been shown to prolong survival in a randomized trial. This concept may be obsolete, as preliminary results for a number of approaches, mostly derived from laboratory observations, show that prostate cancers are not as resistant to chemotherapy as traditionally believed. The population of early "geriatric" HRPC patients is rapidly increasing, posing an even greater challenge to oncologists in coping with this difficult-to-manage patient population. In this article, we analyze the most novel chemotherapeutic combinations for the treatment of HRPC in otherwise healthy elderly men. PMID- 10482186 TI - Immunologic approaches to the treatment of prostate cancer. AB - The presence of several organ-specific molecules that could serve as immunogens or targets of an immune attack, the nonessential nature of the prostate gland, the substantial failure rate after treatment of the primary tumor, and the lack of effective chemotherapy for metastatic disease make prostate cancer an ideal candidate for immunotherapy. This report reviews the current status of two novel approaches to the treatment of prostate cancer. The first is an effort to induce antitumor immunity by enriching the cytokine environment within the primary cancer by intraprostatic injection of Leukocyte Interleukin (Cel-Sci Corp, Vienna, VA), a mixture of natural cytokines that includes interleukin-1 beta (IL 1beta), IL-2, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-gamma), and tumor necrosis factor alpha (TNF-alpha). The second approach uses OncoVax-P (Jenner Biotherapies, Inc, San Ramon, CA), a vaccine consisting of liposome-encapsulated recombinant prostate-specific antigen (PSA) and lipid A. When administered as an emulsion or in association with bacillus Calmette-Guerin (BCG)/cyclophosphamide or GM-CSF with or without IL 2/cyclophosphamide, immunologic tolerance is broken as evidenced by the generation of humoral and cellular immunity. Both of these approaches have been shown to be feasible and safe, and are now being tested in patients with less advanced disease to determine if manipulation of the immune system can favorably influence clinical outcome. PMID- 10482188 TI - Gene therapy for prostate cancer. AB - Prostate cancer is one of the leading causes of cancer deaths in the western hemisphere. A number of different gene therapy strategies are currently being evaluated. The ex vivo and many of the in vivo therapies involve stimulating a specific antitumor immune response. Autologous vaccines involving interleukin-2 (IL-2)- or granulocyte-macrophage colony-stimulating factor (GM-CSF)-transduced whole tumor cells showed great promise in animal models. Clinical trials of these and other vaccine strategies are underway. In vivo gene therapies involving the replacement of mutant tumor-suppressor genes, antisense strategies, and the insertion of suicide genes are also being evaluated in prostate cancer. PMID- 10482187 TI - Peptide and carbohydrate vaccines in relapsed prostate cancer: immunogenicity of synthetic vaccines in man--clinical trials at Memorial Sloan-Kettering Cancer Center. AB - Men with rising prostate-specific antigen (PSA) levels after primary therapies such as prostatectomy or radiotherapy represent a unique group for whom no standard treatment option exists. A variety of approaches including expectant monitoring, dietary modification, hormonal therapy, and alternative medicines have shown an impact on the rate of increase in PSA, but the overall effect on survival remains controversial. At Memorial Sloan-Kettering Cancer Center, we have focused our treatment approach on this cohort of patients in a series of phase I monovalent carbohydrate and glycoprotein-conjugate vaccine trials using the patients' immune system to generate an antitumor response. These synthetic vaccines are conjugated to keyhole limpet hemocyanin (KLH) and given with the immunologic adjuvant QS21 as five subcutaneous vaccines over 26 weeks. All patients generated specific high-titer immunoglobulin M (IgM) and/or IgG antibodies, some of which were able to mediate complement lysis. Preliminary data suggest that these vaccines may impact on the rate of increase in posttreatment PSA slopes compared with pre-PSA values. The impact of vaccine therapy on the PSA slope and its effect on the time to radiographic progression are the current focus of a forthcoming phase II trial. Vaccines may offer an alternative treatment option for the patient who has relapsed early following primary therapies. PMID- 10482189 TI - Current management of metastatic breast cancer. AB - The medical approach to the management of metastatic breast cancer has changed through the years following the introduction of new cytotoxic agents and the demonstration of their antitumor activity. In general, combination drug regimens are the initial treatments of choice when chemotherapy is indicated for patients with metastatic breast cancer. However, the single-agent activity of some of the newer agents rivals that of older combination chemotherapy treatments. Compared with older therapies, improved objective response rates and/or improved duration of response recently have been reported in studies evaluating optimal dosing and sequencing of these newer agents, alone or in combination with other drugs. Although there is no rigid standard for the sequencing of therapy for management of metastatic breast cancer, chemotherapy has a role in the treatment program for nearly all patients with this disease. The goal of treatment remains meaningful palliation of patients with complications of progressive cancer. PMID- 10482190 TI - Recent developments in breast cancer therapy. AB - Over the past three decades conceptual approaches to breast cancer have led to improvements in locoregional therapy and early diagnosis. Systematic screening programs with mammography reduce disease-specific mortality by 25% to 30%, while many patients with early breast cancer receive optimal breast-conserving treatments. Our increased understanding of the biology of breast cancer helped develop successful adjuvant systemic therapies (cytotoxic and hormonal) that, in turn, reduce mortality by 15% to 25%. Newer therapeutic interventions are under intensive investigation. While continued progress in cytotoxic therapy is evident (taxanes, vinorelbine, gemcitabine, new antifolates, liposomal anthracyclines, etc), there is increasing interest in targeting growth factors and their receptors. Thus, a monoclonal antibody directed to the extracellular domain of the HER-2/neu oncoprotein was recently approved by the Food Drug Administration based on evidence of antitumor activity as a single agent and in combination with cytotoxic therapy. A similar approach against the epidermal growth factor receptor is under evaluation in clinical trials. Various methods of inhibiting intracellular signal transduction also are in clinical development. These include tyrosine kinase inhibition, dominant negative mutant inhibitors of GRB-2, farnesyl transferase inhibition and vaccines directed against various epitopes expressed by mammary cancer cells. Angiogenesis and the enzyme telomerase are other targets under intense scrutiny since they are integrally involved with metastases and cellular immortality, both common characteristics of the malignant cell. These lines of investigation are likely to provide innovative therapeutic interventions, which may improve the specificity and therapeutic index of anticancer treatments. PMID- 10482191 TI - Recent advances in adjuvant therapy of breast cancer. AB - Systemic adjuvant therapies that reduce the mortality in breast cancer include ovarian ablation, tamoxifen, and combination chemotherapies. Tamoxifen is effective in reducing the risk of recurrence and death when administered to patients with estrogen receptor-positive tumors. The benefit of tamoxifen was significant, irrespective of age, menopausal status, and whether tamoxifen was given alone or in association with chemotherapy. For patients who had estrogen receptor-poor tumors, there was no significant improvement in disease-free or overall survival. Adjuvant tamoxifen therapy was also associated with a proportional reduction in the incidence of contralateral breast cancer. Anthracycline-containing combination chemotherapy is more effective than other regimens without anthracyclines. Additional alternate treatment with paclitaxel as adjuvant therapy can further reduce the risk of recurrence and improve survival. High-dose chemotherapy remains experimental, and results of ongoing studies will determine its role in the management of primary breast cancer. Escalating doses beyond conventional established doses of currently used chemotherapy drugs does not result in further reduction in risk of recurrence, but adds substantial morbidity as demonstrated by an increased risk of leukemia and myelodysplastic syndrome. PMID- 10482192 TI - Biological therapies for breast carcinoma: concepts for improvement in survival. AB - Systemic treatments for breast carcinoma have improved substantially over the past quarter century. New insights into cancer biology, refinements in biotechnology, and bioengineering of macromolecules hold the promise of even greater reductions in breast and other cancer mortality as a result of biologicals. As exemplified by the clinical results with the monoclonal antibody to HER-2 for antigen-specific passive immunotherapy, biological therapies for breast carcinoma hold substantial promise. The objective of this report is to highlight aspects of preclinical and clinical research on other biologicals for breast carcinoma that also hold potential for improving patient survival. As examples of the potential of cytokines to modulate breast carcinoma cell proliferation and tumor growth, data on cytokines (interferons) with pleiotropic effects and a lymphokine (interleukin-10) acting on T cells and macrophages will be reviewed. HER-2 has promise as a vaccine for active specific immunotherapy; these data will be summarized. Progress on these and other biologicals promises that this will be another modality of therapy resulting in improved survival for patients with both early and metastatic breast carcinoma in the next millennium. PMID- 10482193 TI - An overview of monoclonal antibody therapy of cancer. AB - Monoclonal antibody-based therapeutics are beginning to realize the promise that was predicted with the advent of the core technology more than 20 years ago. Antibody-based therapeutics targeting tumor cell surface antigens such as B-cell idiotypes, CD20 on malignant B cells, CD33 on leukemic blasts, and HER2/neu on breast cancer cells have shown efficacy in clinical trials. Multiple antibody based strategies have shown promising efficacy in recent clinical trials. Unconjugated immunoglobulins directed against CD20 induce partial and complete responses in up to 50% of patients with advanced, indolent non-Hodgkin's lymphoma When such antibodies are conjugated to appropriate radionuclides and administered in therapeutic doses, the proportions of complete and overall responses increase considerably. Conjugates composed of anti-CD33 antibodies and the chemotherapy agent, calicheamicin, show promising activity in patients with relapsed or refractory acute myelogenous leukemia. Treatment of patients with advanced breast cancer using the anti-HER2/neu antibody trastuzumab (Herceptin; Genentech, San Francisco) leads to objective responses in some patients whose tumors overexpress the HER2/neu oncoprotein. These exciting results justify recent enthusiasm for continued efforts to refine existing approaches and to develop new antibody-based strategies to treat human malignancy. PMID- 10482194 TI - Basic science of HER-2/neu: a review. AB - The HER-2/neu (also known as c-erbB-2) oncogene is the second member of the epidermal growth factor receptor family. It is overexpressed in many different types of human cancers, including breast, ovarian, lung, gastric, and oral cancers. Overexpression of HER-2/neu in breast cancer has been associated with poor overall survival and has been shown preclinically to enhance malignancy and the metastatic phenotypes. Although discrepancies exist between different studies, HER-2/ neu overexpression seems to induce chemoresistance in certain experimental conditions. Many studies have convincingly shown that repression of HER-2/neu suppresses the malignant phenotypes of HER-2/neu-overexpressing cancer cells. These findings strongly suggest that HER-2/neu may serve as an excellent target for developing anticancer agents specific for HER-2/neu-overexpressing cancer cells. HER-2/neu-encoded p185 protein is a receptor tyrosine kinase that can be associated with multiple signal transduction pathways. However, it is not yet clear how a specific signal pathway may correspond to a specific biological response. This report reviews basic information on signal transduction of HER 2/neu receptor tyrosine kinase and summarizes our approaches to targeting HER 2/neu-overexpressing cancer cells. The HER-2/neu promoter was targeted using cationic liposomes or an adenovirus vector to deliver the adenovirus-5 EIA gene products and a nontransformed mutant of the SV40 large T antigen into the tumor bearing mice. This resulted in suppression of the tumor growth and prolongation of survival. For repressing the function of HER-2/neu we used emodin, a tyrosine kinase inhibitor. This agent can inhibit the tyrosine kinase activity of HER 2/neu and preferentially block the growth of the HER-2/neu-overexpressing human breast cancer cells in tissue culture as well as in nude mice. PMID- 10482195 TI - Nonclinical studies addressing the mechanism of action of trastuzumab (Herceptin). AB - HER2 is a ligand-less member of the human epidermal growth factor receptor or ErbB family of tyrosine kinases. In normal biological systems, HER2 functions as a co-receptor for a multitude of epidermal growth factor-like ligands that bind and activate other HER family members. HER2 overexpression is observed in a number of human adenocarcinomas and results in constitutive HER2 activation. Specific targeting of these tumors can be accomplished with antibodies directed against the extracellular domain of the HER2 protein. One of these antibodies, 4D5, has been fully humanized and is termed trastuzumab (Herceptin; Genentech, San Francisco, CA). Treatment of HER2-overexpressing breast cancer cell lines with trastuzumab results in induction of p27KIP1 and the Rb-related protein, p130, which in turn significantly reduces the number of cells undergoing S-phase. A number of other phenotypic changes are observed in vitro as a consequence of trastuzumab binding to HER2-overexpressing cells. These phenotypic changes include downmodulation of the HER2 receptor, inhibition of tumor cell growth, reversed cytokine resistance, restored E-cadherin expression levels, and reduced vascular endothelial growth factor production. Interaction of trastuzumab with the human immune system via its human immunoglobulin G1 Fc domain may potentiate its antitumor activities. In vitro studies demonstrate that trastuzumab is very effective in mediating antibody-dependent cell-mediated cytotoxicity against HER2 overexpressing tumor targets. Trastuzumab treatment of mouse xenograft models results in marked suppression of tumor growth. When given in combination with standard cytotoxic chemotherapeutic agents, trastuzumab treatment generally results in statistically superior antitumor efficacy compared with either agent given alone. Taken together, these studies suggest that the mechanism of action of trastuzumab includes antagonizing the constitutive growth-signaling properties of the HER2 system, enlisting immune cells to attack and kill the tumor target, and augmenting chemotherapy-induced cytotoxicity. PMID- 10482196 TI - Overview of the trastuzumab (Herceptin) anti-HER2 monoclonal antibody clinical program in HER2-overexpressing metastatic breast cancer. Herceptin Multinational Investigator Study Group. AB - The recombinant humanized anti-HER2 monoclonal antibody trastuzumab (Herceptin; Genentech, San Francisco, CA) was evaluated in human clinical trials for treatment of women with metastatic breast cancer who have tumors that overexpress HER2. The trastuzumab clinical program consisted of a series of phase I, phase II, and phase III clinical trials. Clinical experience with this novel biologic has been obtained in more than 1,000 women with HER2-overexpressing metastatic breast cancer. Two pivotal trials were performed to evaluate trastuzumab efficacy and safety: (1) trastuzumab in combination with chemotherapy as first-line therapy and (2) trastuzumab as a single agent in second- and third-line chemotherapy. Preliminary results of the pivotal clinical trials that have been presented at national meetings are summarized below. The data suggest that trastuzumab will be an important new treatment option for women with HER2 overexpressing metastatic breast cancer. PMID- 10482197 TI - Phase II study of weekly intravenous trastuzumab (Herceptin) in patients with HER2/neu-overexpressing metastatic breast cancer. AB - The HER2/neu proto-oncogene is overexpressed in 25% to 30% of patients with breast cancer. Trastuzumab (Herceptin; Genentech, San Francisco, CA), a recombinant humanized monoclonal antibody with high affinity for the HER2 protein, inhibits the growth of breast cancer cells overexpressing HER2. In this phase II study the efficacy and toxicity of weekly administration of trastuzumab was evaluated in 46 patients with metastatic breast cancer whose tumors overexpressed HER2. A loading dose of 250 mg trastuzumab was administered intravenously, which was followed by 10 weekly doses of 100 mg each. Upon completion of this treatment period, patients with no disease progression could receive a weekly maintenance dose of 100 mg. Patients in this trial had extensive metastatic disease, and most had received prior anticancer therapy. Ninety percent of patients achieved adequate serum levels of trastuzumab. Toxicity was minimal, and no antibodies against trastuzumab could be detected. Objective responses were observed in 5 of the 43 evaluable patients, which included 1 complete remission and 4 partial remissions, for an overall response rate of 11.6%. Responses were seen in mediastinum, lymph nodes, liver, and chest wall lesions. Minor responses (seen in 2 patients) and stable disease (14 patients) lasted for a median of 5.1 months. These results demonstrate that trastuzumab is well tolerated and clinically active in patients with HER2-overexpressing metastatic breast cancers who have received extensive prior therapy. The regression of human cancer through the targeting of putative growth factor receptors such as HER2 warrants further evaluation of trastuzumab in the treatment of breast cancer. PMID- 10482198 TI - Health-related quality of life in women with metastatic breast cancer treated with trastuzumab (Herceptin). AB - The measurement of health-related quality of life (HRQL) was an important component of clinical trials of trastuzumab (Herceptin; Genentech, San Francisco, CA) in women with progressive HER2-overexpressing metastatic breast cancer who may or may not have had prior chemotherapy. Health-related quality of life was measured at baseline and specified intervals during therapy using the European Organization for Research and Treatment of Cancer core Quality of Life Questionnaire (QLQ-C30, version 1.0). Five domains were chosen a priori for analysis: global quality of life, physical, role and social functioning, and fatigue. In the phase II study, in which trastuzumab was given alone, there was no change in on-treatment QLQ-C30 scores compared with baseline. These results suggest that trastuzumab does not adversely affect HRQL during therapy. In the phase III study, the effects of trastuzumab plus chemotherapy were compared with those of chemotherapy alone. A comparison of QLQ-C30 scores during treatment did not show statistically significant differences between the two groups at the preset level of P = .01. However, comparison of on-treatment scores with baseline in patients receiving chemotherapy alone indicated mild worsening of physical and role functioning and of fatigue throughout the duration of treatment, whereas a similar comparison of those receiving chemotherapy with trastuzumab revealed mild worsening of role functioning at weeks 8 and 20 and of fatigue only at week 8. These results suggest that trastuzumab may be associated with an amelioration of the deleterious effects of chemotherapy alone. In summary, in the doses and schedules used in these studies, trastuzumab is not associated with worsening of HRQL. PMID- 10482199 TI - Combination therapy with trastuzumab (Herceptin) and cisplatin for chemoresistant metastatic breast cancer: evidence for receptor-enhanced chemosensitivity. AB - The anti-HER-2/neu antibody trastuzumab (Herceptin; Genentech, San Francisco, CA) interferes with DNA repair induced by cisplatin and, as a result, promotes cytotoxicity in HER-2/neu-overexpressing tumor target cells in a synergistic fashion. This effect of trastuzumab, termed receptor-enhanced chemosensitivity, is specific for HER-2/neu-overexpressing cells, having no effect on cells without overexpression. Based on these findings, we conducted phase I and II clinical trials of trastuzumab plus cisplatin to determine the toxicity, pharmacokinetics, response rate, and response duration of this combination in patients with HER 2/neu-overexpressing metastatic breast cancer who had demonstrated disease progression (chemoresistance) while on active chemotherapy just prior to study entry. In phase I, four of 15 patients had objective clinical responses, including one complete response of several years' duration. Of 37 assessable patients enrolled in phase II, nine (24.3%) had objective clinical responses and an additional nine had minor responses or stable disease. The median time to progression among the responders was 8.4 months. The toxicity profile reflected that expected from cisplatin alone, with no apparent increase in toxicity caused by the addition of trastuzumab. Moreover, the pharmacokinetics of trastuzumab were unaltered by coadministration of cisplatin. We conclude that the combination of trastuzumab and cisplatin results in response rates higher than that reported for either single agent alone. Such receptor-enhanced chemosensitivity offers a new approach to target overexpressed growth factor receptors in a variety of cancers, which will lead to new, biologically based therapeutic strategies for clinical intervention. PMID- 10482200 TI - Cardiotoxicity in patients receiving transtuzumab (Herceptin): primary toxicity, synergistic or sequential stress, or surveillance artifact? AB - Human epidermal growth factor receptor-2 (HER2) is a member of the epidermal growth factor receptor family, which produces factors that are considered to be important mediators of cell growth. Overexpression of HER2, which occurs in approximately 25% to 30% of human breast cancers, has fostered considerable interest in innovative therapeutic modalities designed to target tumor cells demonstrating such overexpression. Trastuzumab (Herceptin; Genentech, San Francisco, CA), a humanized monoclonal antibody developed to target the HER2 receptor, is the most widely studied example of such a modality. In early clinical studies with trastuzumab, cardiomyopathy was observed with a clinical expression similar to that seen with the anthracyclines (ie, a potentially progressive decrease in cardiac systolic function). A number of possible explanations for this cardiotoxicity are explored in this report. The first is that trastuzumab has inherent toxicity. This consideration has some theoretical interest, since fetal myocardial cells exhibit HER2 receptors and the adult myocardium expresses HER3 receptors. A second possibility is that sequential stresses following doxorubicin administration contribute to cardiac dysfunction. A third explanation is that observational artifacts lead to an overestimation of trastuzumab cardiotoxicity. Approaches for additional study of the extent and severity of trastuzumab cardiotoxicity are briefly addressed. PMID- 10482201 TI - Issues in the design of clinical trials: insights from the trastuzumab (Herceptin) experience. AB - A guiding objective of phase III clinical trials should be to achieve reliable evaluation of safety and efficacy of interventions in a real-world setting, ie, as the interventions would be delivered in clinical practice. This guiding objective motivates several principles of design. The control regimen should truly reflect standard of care; in turn, one should be reluctant to use placebos that might not be inert or that might adversely impact quality of life. The choice of concomitant therapy should be sufficiently flexible as to allow for treatment options that would be widely representative of routine care. Eligibility criteria should be as inclusive as the intended labeling, specifically excluding only those patients expected to be at high risk for toxicity or have a low likelihood of benefit. Study procedures should be simplified to avoid unnecessary inconvenience to patients and investigators and to reduce trial costs. Clinical end points should be chosen that unequivocally reflect tangible benefit to patients. Experiences from the phase III trial of trastuzumab (Herceptin; Genentech, San Francisco, CA) in metastatic breast cancer, a human immunodeficiency virus/acquired immunodeficiency syndrome trial, and a trial in cardiology will be used to provide additional motivation for several of these principles. These trials provide valuable insights into the challenging issues arising in the design and conduct of clinical trials. PMID- 10482202 TI - The role of immunohistochemistry and fluorescence in situ hybridization for HER2/neu in assessing the prognosis of breast cancer. AB - Immunohistochemistry has been used in the past to identify overexpression of HER 2/neu protein on the cell membrane of breast cancer cells in fixed tissues. Most studies that have attempted to find an association between HER-2/neu expression and a poor prognosis have relied on this technique, which has intrinsic variability due to the antibody used and the degradation of surface proteins by fixation. Recent studies with fluorescence in situ hybridization have tended to confirm the purported association, showing that HER-2/neu overexpression causes a more aggressive, less responsive breast cancer. In many studies, the amplification of the HER-2/neu gene was the most important variable determining outcome, independent of other variables, such as tumor size and estrogen receptor status. PMID- 10482203 TI - Should HER2 status be routinely measured for all breast cancer patients? AB - Several recent publications have re-opened the question of whether HER2 status should be determined for all patients with newly diagnosed breast cancer. The barrier in the past to the use of HER2 has been the nonstandardization of HER2 status determination, which is the major caveat to its use today. Two test kits have been recently approved by the Food and Drug Administration for HER2 testing, one for determining HER2 amplification by fluorescence in situ hybridization and the other for measuring HER2 overexpression by immunohistochemistry. Neither of these tests, nor any of the other myriad tests used for HER2 determinations, has been validated in all the potential arenas for the use of HER2: refinement of estimates of prognosis of untreated low-risk patients, selection among treatment options for adjuvant therapy, and selection of patients for treatment with trastuzumab (Herceptin; Genentech, San Francisco, CA). The nonstandardization of testing has led to conflicting results and controversy as to the value of HER2 in evaluating breast cancer patient prognosis and the selection among therapeutic options. Thus, testing for HER2 is not yet routine for patients with newly diagnosed breast cancer, although it is of value for patients who develop metastatic disease and who need to know if they are candidates for trastuzumab. PMID- 10482204 TI - A commentary on the use of rational oral electrolyte therapy. PMID- 10482205 TI - A chronology of pain and comfort in children with sickle cell disease. AB - OBJECTIVE: To examine the patterns of children's and caregivers' descriptions of pain and the comfort measures used to relieve the pain of sickle cell disease (SCD) at home and in the hospital. DESIGN: Qualitative and quantitative techniques were used to obtain data. Participants were seen twice, first with and then without a vaso-occlusive episode. Multiple simultaneous methodological triangulation was used to integrate the findings from ethnographic interviews and observations as well as limited quantitative findings about pain and comfort measures used. SETTING: A Midwestern children's hospital with a regional SCD service. PARTICIPANTS: Twenty-one African American children and adolescents with SCD, aged 6 to 15 years, and 21 family caregivers. RESULTS: An 8-phase chronology of pain and comfort was revealed from the data about pain and comfort in children with SCD. Although this chronology was an unexpected finding, it was consistent with the original aim of the investigation. Phase 1 (baseline) represented the usual state of the child's condition, which for most was free of pain. Phase 2 or the "pre-pain" state involved no vasoocclusive pain but the child began to show prodromal signs and symptoms of painful episodes, such as yellowing of the eyes or fatigue. Phases 2 through 7 involved increasing then decreasing levels of pain, including the pain start point (phase 3), pain acceleration (phase 4), peak pain experience (phase 5), pain decrease start point (phase 6), and steady pain decline (phase 7). A trip to the emergency department usually occurred during phase 5. In phase 8 (pain resolution), the pain had decreased to a manageable level so that the child could be discharged from the hospital. As pain increased and decreased, so did the number and variety of comfort measures. CONCLUSIONS: A chronology of the pain and comfort experiences for children and adolescents during a vasoocclusive event of SCD emerged from the descriptive data of this study. Findings need to be examined further in larger, quantitative, longitudinal studies that examine more closely the duration, intensity, and character of pain at different times during vaso-occlusive episodes as well as the comfort measures used during specific phases of the pain event. PMID- 10482206 TI - The role of serial radiographs in the management of pediatric torus fractures. AB - OBJECTIVES: To assess the utility of radiographs taken immediately after the application of a cast in the management of pediatric torus (or buckle) fractures and to determine the need for serial radiographs taken at follow-up visits. DESIGN: Retrospective medical record review; survey questionnaire of a panel of experts. SETTING: The pediatric emergency department (PED) and the pediatric orthopedic clinic at an urban, tertiary care hospital. PATIENTS: All children with torus fractures referred to the pediatric orthopedic clinic for follow-up visits between February 1995 and February 1997. MAIN OUTCOME MEASURES: The number of patients whose postcast studies was obtained in the PED; number of follow-up visits and studies conducted at the pediatric orthopedic clinic; usual regional practices as extracted from a panel of experts by survey questionnaire. RESULTS: Of 70 patients, 46 (66%) were evaluated by a single, precast study in the PED, and 24 (34%) were evaluated by both precast and postcast studies in the PED. The time range of the first follow-up study was the first to fifth week after the patient's injury. The range of the number of follow-up studies for each patient was 0 to 5. Our cohort's total radiology charges for 70 patients were $27251. Regional directors of pediatric orthopedic surgery unanimously agreed that postcast studies in the PED are unnecessary. The range of the number of follow-up studies they obtained is 0 to 3 per patient. CONCLUSIONS: Postcast studies of torus fractures are unnecessary. Multiple radiographs taken during follow-up visits, especially early in the healing process, do not change fracture management. Relying on the clinical examination, perhaps combined with a single follow-up study, is a more appropriate regimen for the management of pediatric torus fractures and translates into a cost savings of over $ 10000 for our 70 patients. PMID- 10482207 TI - Maternal disciplinary practices in an at-risk population. AB - OBJECTIVES: To describe maternal discipline of children in at-risk families and to determine factors associated with disciplinary practices. DESIGN: Cross sectional survey. SETTING: At-risk families in North Carolina followed up in a longitudinal study of child maltreatment. PARTICIPANTS: Maternal caregivers of 7- to 9-year-old children with factors at birth that placed them at risk. MEASUREMENTS AND RESULTS: A total of 186 maternal caregivers were interviewed. A measure, based on coding parental responses, was used to assess disciplinary practices for 5 different misbehaviors. Limit setting was the most commonly used disciplinary practice for 4 of 5 misbehaviors, with 63% of mothers reporting that this method generally worked best. Spanking was more likely used as a secondary response for each misbehavior, when the primary one had not succeeded. Conversely, teaching or verbal assertion was always less likely as a secondary response. Teaching or verbal assertion was used more commonly for lying than for any other misbehavior, limit setting for disobeying, spanking for stealing, and spanking with an object for being disrespectful. Regression modeling for the 4 most common disciplinary practices showed (P<.05) that black race, lack of Aid to Families With Dependent Children receipt, more-educated mothers, and female sex of child were associated with higher use of teaching or verbal assertion; a biological father in the home was associated with less use of limit setting; and black race and report for child maltreatment were associated with more use of mild spanking. CONCLUSIONS: In this sample, limit setting was the most common disciplinary technique. Disciplinary practices used varied depending on the type of misbehavior and other contextual factors, including child, parent, and family characteristics. PMID- 10482208 TI - The tobacco industry and underage youth smoking: tobacco industry documents from the Minnesota litigation. AB - OBJECTIVE: To learn how underage youth have been regarded by the major tobacco companies, as revealed in industry documents during the Minnesota litigation. DATA SOURCES: Documents from Brown & Williamson Tobacco Corp, Lorillard Tobacco Co, Philip Morris Inc, and R. J. Reynolds Tobacco Co. STUDY SELECTION: Documents available from the State of Minnesota and Blue Cross and Blue Shield of Minnesota v Philip Morris Inc et al during the discovery process of the trial. Hundreds of industry documents related to youth, teenagers, and young adults were examined. A sample of documents are referenced as illustrations of key points. CONCLUSIONS: Underage youths are viewed by the tobacco industry as critical to its future. Underage youth smoking patterns were examined for decades. Reductions in youth smoking rates were seen by tobacco companies as a negative trend for the companies. Specific marketing campaigns were directed at underage youth and were successful with this age group. Tobacco industry practices should continue to be carefully scrutinized. PMID- 10482209 TI - Meyer dysplasia in the differential diagnosis of hip disease in young children. AB - OBJECTIVES: To describe a rare developmental disorder of the femoral capital epiphysis in infants and children that is often misdiagnosed and to suggest an evaluation protocol to differentiate it from other hip problems. DESIGN: Case series. SETTING: Tertiary care center. SUBJECTS: Five consecutive patients referred for evaluation of acute onset of limping between January 1990 and December 1997. INTERVENTION: All clinical and imaging data were collected. RESULTS: Two of the 5 patients were initially diagnosed as having osteomyelitis and 3 as having Perthes disease. The diagnosis of Meyer dysplasia was confirmed by plain film of the pelvis, a negative bone scan, or normal bone marrow findings on magnetic resonance imaging. The limping resolved without treatment in all patients within 1 to 3 weeks. CONCLUSIONS: Meyer dysplasia is a benign condition that should be included in the differential diagnosis of hip disease in infants and children. Awareness of this condition may prevent unnecessary hospitalization and treatment. PMID- 10482211 TI - The role of transthoracic echocardiography in the diagnosis of infective endocarditis in children. AB - BACKGROUND: Infective endocarditis (IE) is frequently suspected but infrequently diagnosed in children. Clinicians often order echocardiograms to "rule out" IE. In an era of cost constraint, clinically efficient strategies must be developed to eliminate unnecessary tests. We hypothesized that transthoracic echocardiography (TTE) is only useful in children in whom there is a high clinical suspicion of IE based on history, physical examination, and persistently positive blood cultures. OBJECTIVE: To determine the role of TTE as a screening test for suspected IE in children. METHODS: Echocardiographic reports and medical records were reviewed retrospectively for 173 consecutive patients who underwent TTE to rule out IE from January 1993 to August 1996. RESULTS: Persistent fever was the predominant symptom leading to a suspicion of IE (120 patients [69.4%]). Fifty-seven (32.9%) of the 173 patients had congenital heart disease and 95 patients (54.9%) had indwelling venous catheters. Twenty-six patients (15.0%) were diagnosed and treated for IE. Twelve (46.2%) of these 26 patients had vegetations seen on TTE. The conditions of the remaining 14 patients were diagnosed clinically and these patients had persistently positive blood cultures. By univariate analysis, the risk factors associated with the diagnosis of IE were malaise, congestive heart failure, new or changing heart murmur, leukocytosis, hematuria, and the presence of 2 or more positive blood cultures for the same organism. The risk factors associated with positive TTE were malaise, congestive heart failure, new or changing heart murmur, leukocytosis, hematuria, and 2 or more positive blood cultures. The presence of an indwelling catheter or immunocompromised status were not predictive of vegetation or IE. CONCLUSIONS: Transthoracic echocardiography has poor sensitivity as a screening test for IE in patients with low clinical probability of the disease. A diagnostic algorithm for IE is suggested based on these data. PMID- 10482210 TI - Seizures decrease rapidly after fasting: preliminary studies of the ketogenic diet. AB - OBJECTIVES: To evaluate the change in atonic or myoclonic seizures associated with the Lennox-Gastaut syndrome during the initiation of the ketogenic diet, and to describe the development of a blinded crossover study of the efficacy of the ketogenic diet. DESIGN: A before-after trial. SETTING: The Johns Hopkins Hospital, Baltimore, Md. PATIENTS: Change in clinical seizure frequency was examined in 17 consecutively treated patients with atonic or myoclonic seizures. In a few patients, a 24-hour ambulatory electroencephalogram was obtained before and after diet initiation. We demonstrated the ability to manipulate the ketosis induced by fasting with the addition of glucose (dextrose) in 1 patient. INTERVENTIONS: Children fasted for 36 hours, and the diet was gradually introduced over 3 days. Parents were instructed to keep a baseline seizure frequency calendar for the month before the initiation of the diet. These calendars continued to be maintained as the diet was initiated. MAIN OUTCOME MEASURE: Seizure decrease from baseline. RESULTS: The atonic or myoclonic seizures decreased in these children by more than 50% immediately. Using a 24 hour ambulatory electroencephalogram, we documented that the seizures reported by a parent represent only a fraction of the electroclinical events; the technique could be used to measure the profound decrease in electrically documented seizures. Ketosis was eliminated with glucose, 60 g/d. CONCLUSIONS: It is feasible to evaluate the ketogenic diet's efficacy in atonic or myoclonic seizures in a blinded, crossover study. The diet can be manipulated on a short term basis in a blinded manner, and ketosis can be achieved or eliminated. PMID- 10482212 TI - Family presence during invasive procedures in the pediatric intensive care unit: a prospective study. AB - OBJECTIVES: To determine if allowing 1 or both parents to be present during invasive procedures reduces the anxiety that parents experience while their child is in the pediatric intensive care unit; to evaluate if the parent's presence was helpful to the child and parent; and to determine whether this presence was harmful to the nurses or physicians. DESIGN: A prospective study using surveys (5 point Likert scale) of parents of children requiring intubation, placement of central lines, or chest tubes. Additional surveys were completed by bedside nurses to evaluate the effects of parental presence. SETTING: A 12-bed pediatric intensive care unit in upstate New York. PARTICIPANTS: The study population consisted of the parents of 16 children undergoing 1 or more procedures; 7 had undergone intubation, 11 had central lines placed, and 2 had chest tubes placed. The control population consisted of the parents of 7 children undergoing 1 or more procedures; 7 had undergone intubation, 5 had central lines placed, and 3 had chest tubes placed. RESULTS: Parental presence significantly reduced the parental anxiety related to the procedure (P = .005; Mann-Whitney test), but did not change condition-related anxiety (P = 0.9; Mann-Whitney test). Thirteen of 16 parents found their presence helpful to themselves (10 very, 3 somewhat) and the medical staff (11 very); 14 of 16 found their presence helpful to their child (11 very). Fifteen (94%) of 16 parents would repeat their choice to watch. Fifteen (94%) of 16 nurses found parents' presence helpful to the child (9 very) and to the parents (10 very). One nurse found a parent's presence somewhat harmful to nurses and very harmful to the parent. Thirteen (72%) of 18 nurses indicated that allowing parents to observe procedures was an appropriate policy. There were no significant differences noted in response of nurses based on years of experience. CONCLUSIONS: Allowing parental presence during procedures decreases procedure related anxiety. The implications of such a policy change on physicians and other aspects of pediatric intensive care, including medical education, need further evaluation. PMID- 10482213 TI - Presenting the facts about smoking to adolescents: effects of an autonomy supportive style. AB - OBJECTIVES: To test the self-determination model of health-related behavior by examining whether the degree to which adolescents experience an appeal to not smoke as autonomy supportive would affect their autonomous motivation for not smoking and, in turn, their behavior of either refraining from smoking or smoking less, and to validate the measures of perceived autonomy support and autonomous motivation for not smoking. DESIGN: Two studies of physicians presenting information about not smoking using 2 message styles, 1 of which was designed to be more autonomy supportive. The preliminary study involved nonrandomized assignment to message style and only immediate assessment of perceptions, motivation, and behavior, while the primary study involved randomized assignment and 4-month longitudinal assessments. SETTING AND PARTICIPANTS: Nearly 400 ninth- through 12th-grade students at 2 suburban high schools in upstate New York. MAIN OUTCOME MEASURES: Adolescents' perceptions of the presentations' autonomy supportiveness of the presenters, as well as adolescents' autonomous motivation for not smoking and their self-reports of smoking. The primary study also assessed change in students' autonomous motivation and change in their self reported smoking during 4 months. RESULTS: In both studies, the measures were reliable and valid. Students perceived significantly (P = .04 and P<.001, respectively) greater autonomy support in the "It's Your Choice" presentation, after controlling for whether the students were smokers. Perceived autonomy supportiveness of the presentation was positively correlated with autonomous reasons for not smoking in the preliminary study and with increases in autonomous motivation for not smoking in the primary study. Change in autonomous reasons for not smoking significantly (P<.001) predicted reduction in smoking during 4 months. CONCLUSION: When adolescents perceived messages about not smoking as autonomy supportive, they had more autonomous motivation for not smoking, and that, in turn, predicted a decrease in their self-reports of smoking. PMID- 10482214 TI - Cost-effectiveness of laparoscopy in children. AB - BACKGROUND: Laparoscopy may offer fast recovery and improved cosmesis, but its cost has been perceived as excessive. OBJECTIVE: To analyze the total hospital costs of laparoscopy vs open surgery. DESIGN: Retrospective cost-effectiveness analysis evaluating all cases performed in a 36-month period (September 1995 to August 1998). Cases were evaluated for operative time, itemized cost of supplies, and length of hospitalization. SETTING: Operations performed by pediatric surgeons in a tertiary care children's hospital. PATIENTS: Consecutive children undergoing laparoscopic or open appendectomies, cholecystectomies, fundoplications, and splenectomies. Patients were not randomized to laparoscopy, or open surgery. INTERVENTIONS: Laparoscopic procedures performed with a core set of reusable equipment and a limited number of disposable instruments. MAIN OUTCOME MEASURES: Cost surplus of laparoscopy was evaluated, and compared with savings associated with decreased hospital stay, to obtain cost-effectiveness of laparoscopy per procedure. RESULTS: There were 26 laparoscopic and 359 open appendectomies; 33 laparoscopic and 3 open cholecystectomies; 16 laparoscopic and 18 open fundoplications; and 16 laparoscopic and 7 open splenectomies. Excess operating costs per procedure were $442.00 for appendectomy, $634.60 for fundoplication, $847.50 for cholecystectomy, and $1551.30 for splenectomy. Hospital stay was decreased for all laparoscopies, resulting in an overall savings per laparoscopic procedure of $2369.90 for appendectomy, $5390.90 for fundoplication, $1161.00 for cholecystectomy, and $858.90 for splenectomy. CONCLUSIONS: Laparoscopy is cost-effective, particularly for fundoplication, appendectomy, and cholecystectomy. Detailing the costs of supplies, operating time, and length of stay allows interinstitutional comparison and critical cost analysis of laparoscopy. With a more selective use of reusable instruments and further shortening of operative time, the global savings of laparoscopy may increase. PMID- 10482215 TI - Differential permethrin susceptibility of head lice sampled in the United States and Borneo. AB - BACKGROUND: Pediculiasis is treated aggressively in the United States, mainly with permethrin- and pyrethrin-containing pediculicides. Increasingly frequent anecdotal reports of treatment failure suggest the emergence of insecticidal resistance by these lice. OBJECTIVE: To confirm or refute the susceptibility of head lice sampled in the United States to permethrin. DESIGN: Survey. Head lice were removed from children residing where pediculicides are readily available and where such products are essentially unknown. Their survival was compared following exposure to residues of graded doses of permethrin in an in vitro bioassay. SETTING: School children from Massachusetts, Idaho, and Sabah (Malaysian Borneo). SUBJECTS: In the United States, 75 children aged 5 to 8 years. In Sabah, 59 boys aged 6 to 13 years. Virtually all sampled US children had previously been treated with pediculicides containing pyrethrins or permethrin; none of the Sabahan children were so exposed. MAIN OUTCOME MEASURE: Survival of head lice exposed to permethrin. RESULTS: Permethrin did not affect head lice sampled from chronically infested US children who had previously been treated for pediculiasis. The slope of the dose-response regression line for these lice did not differ significantly from zero (P = .66). This pediculicide immobilized lice sampled in Sabah. Mortality correlated closely with permethrin concentration (P = .008). CONCLUSIONS: Head lice in the United States are less susceptible to permethrin than are those in Sabah. The pyrethroid susceptibility of the general population of head lice in the United States, however, remains poorly defined. Accordingly, these relatively safe over-the-counter preparations may remain the pediculicides of choice for newly recognized louse infestations. PMID- 10482216 TI - Impact of a school-based interdisciplinary intervention on diet and physical activity among urban primary school children: eat well and keep moving. AB - OBJECTIVE: To evaluate the impact of a school-based interdisciplinary health behavior intervention on diet and physical activity among children in grades 4 and 5. DESIGN: A quasiexperimental field trial with 6 intervention and 8 matched control schools. Outcomes were assessed longitudinally using preintervention (fall 1995) and follow-up (spring 1997) student survey food frequency and activity measures and follow-up 24-hour recall measures of diet and activity. Change was also assessed using yearly repeated cross-sectional surveys of all grade 5 students from 1995 through 1997. PARTICIPANTS: Longitudinal data were collected from 479 students initially in grade 4 in Baltimore, Md, public schools; 91% were African American. Repeated 24-hour recall measures in 1997 were collected for a random subsample of 336 students. Cross-sectional survey data were collected from all grade 5 students in 1995,1996, and 1997 (n = 2103). INTERVENTION: The Eat Well and Keep Moving Program was taught by classroom teachers over 2 years in math, science, language arts, and social studies classes. Materials provided links to school food services and families and provided training and wellness programs for teachers and other staff members. Intervention materials focused on decreasing consumption of foods high in total and saturated fat and increasing fruit and vegetable intake, as well as reducing television viewing and increasing physical activity. MAIN OUTCOME MEASURES: Dietary intake and physical activity measured via repeated 24-hour recall were primary end points, with additional food frequency and activity measures. RESULTS: The 24-hour recall measures indicated that, after controlling for baseline covariates, the percentages of total energy from fat and saturated fat were reduced among students in intervention compared with control schools (-1.4%; 95% confidence interval [CI], -2.8 to -0.04; P = .04 and -0.60%; 95% CI, -1.2 to 0.01; P = .05). There was an increase in fruit and vegetable intake (0.36 servings/4184 kJ; 95% CI, 0.10-0.62; P=.01), in vitamin C intake (8.8 mg/4184 kJ; 95% CI, 2.0-16; P=.01), and in fiber consumption (0.7 g/4184 kJ; 95% CI, 0.0-1.4; P=.05). Television viewing was marginally reduced (-0.55 h/d; 95% CI, -1.04 to 0.04; P=.06). Analysis of longitudinal and repeated cross-sectional food frequency data indicated similar significant decreases in the percentages of total energy from fat and saturated fat. CONCLUSION: Evaluation of the Eat Well and Keep Moving Program indicates effectiveness in improving dietary intake of students and reducing television viewing. PMID- 10482217 TI - Myeloid colony-stimulating factors: use in the newborn. AB - Bacterial and fungal sepsis are major causes of morbidity and mortality in the newborn. Multiple factors contribute to this increased susceptibility to infection, including quantitative and qualitative neutrophil defects, with a reduction in neutrophil number and function. Neutropenia in the newborn may occur in association with sepsis and has a poor prognosis. In addition to antibiotic therapy and supportive care, granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) have been used to reduce morbidity and mortality. Granulocyte CSF is the physiological regulator of neutrophil production and function. Administration of G-CSF results in increased neutrophil production and counts and improved neutrophil function. Several studies of animal and human newborns having neutropenia or suspected sepsis investigated the use of G-CSF and GM-CSF to elevate neutrophil counts and reduce morbidity and mortality in this population. Results of small clinical trials using G-CSF and GM-CSF in very low-birth-weight infants having neutropenia show increased neutrophil counts and a reduced incidence of sepsis during the neonatal period. Despite these promising early results, further studies of the safety and efficacy of G-CSF and GM-CSF administration in neonates are required before their routine use can be recommended as either prophylaxis or treatment for neonatal sepsis. PMID- 10482218 TI - Pediatric clerkship experience and performance in the Nebraska Education Consortium: a community vs university comparison. AB - OBJECTIVE: To compare the reported experiences and performance on end-of-course examinations of students completing their pediatric clerkship at the University of Nebraska Medical Center (UNMC), Omaha, with that of students completing their clerkship in a community pediatrician's practice (CPP) outside the Omaha metropolitan area. DESIGN: Cohort study. SETTING: Private and/or institutional practices with both ambulatory and hospital components. PARTICIPANTS: For the academic year 1996-1997, all 113 students completing the 8-week third-year pediatric clerkship returned a questionnaire detailing their opinions of the experience. They also completed written (multiple-choice and essay questions) and oral (standardized parent interview) examinations, locally prepared and based on clerkship curriculum objectives provided to the students at orientation. Prior to student placement in the CPP, the clerkship goals, content, and evaluation methods as well as techniques for teaching in a busy office practice were reviewed with the CPP physicians. Eighty-one students performed their clerkship at UNMC while 31 spent all but the first week of the clerkship in the CPP. MAIN OUTCOME MEASURES: The students' opinions about their experiences and their performances on the end-of-course examinations were compared. Statistical analysis of the questionnaire was done using the Fisher exact test and the Mantel Haenszel chi2 test while examination performance was compared using the t test and the Wilcoxon rank sum test. RESULTS: The UNMC and CPP groups reported similar opinions of their experiences in the newborn nursery and the inpatient portion of the clerkship, but the CPP students were much more positive about their learning experience in the clinic (P=.001). The CPP students reported more involvement in the patient's overall care (P<.001) and in other aspects of clinic operation (P<.001). The UNMC and CPP students had similar opinions of curriculum content, reading material, and didactic instruction. No group differences were found regarding interest in pediatrics as a career. Most importantly, no group differences were found in performance on any portion of the end-of-course examinations. CONCLUSIONS: Community-based education at the third-year clerkship level can be accomplished without a significant effect on student examination performance if students and faculty are aware of and adhere to a common set of goals. The end result is a much more robust experience for students who spend the clerkship in the practice of a community-based pediatrician. PMID- 10482219 TI - Radiological case of the month. Takayasu arteritis. PMID- 10482220 TI - Picture of the month. Anomalous origin of the left coronary artery. PMID- 10482221 TI - Pathological case of the month. Papillary carcinoma of the thyroid in a thyroglossal duct remnant. PMID- 10482222 TI - Nutrition and jaundice: hold the bottle, keep the breast. PMID- 10482223 TI - Cost of care coordination for children with special health care needs. PMID- 10482224 TI - Narcolepsy and the hypocretin receptor 2 gene. PMID- 10482225 TI - A case of mistaken identity. PMID- 10482226 TI - Creating barriers: a new role for Schwann cells and Desert hedgehog. PMID- 10482227 TI - What's all the RAVE about receptor internalization? PMID- 10482228 TI - The smell of representations. PMID- 10482229 TI - Neuronal polarity: controlling the sorting and diffusion of membrane components. PMID- 10482230 TI - Generation and maintenance of neuronal polarity: mechanisms of transport and targeting. PMID- 10482231 TI - mRNA trafficking and local protein synthesis at the synapse. PMID- 10482232 TI - Neuronal polarity and the epithelial metaphor. PMID- 10482233 TI - Dynamic selectivity filters in ion channels. PMID- 10482234 TI - Requirement for the homeobox gene Hb9 in the consolidation of motor neuron identity. AB - The homeobox gene Hb9, like its close relative MNR2, is expressed selectively by motor neurons (MNs) in the developing vertebrate CNS. In embryonic chick spinal cord, the ectopic expression of MNR2 or Hb9 is sufficient to trigger MN differentiation and to repress the differentiation of an adjacent population of V2 interneurons. Here, we provide genetic evidence that Hb9 has an essential role in MN differentiation. In mice lacking Hb9 function, MNs are generated on schedule and in normal numbers but transiently acquire molecular features of V2 interneurons. The aberrant specification of MN identity is associated with defects in the migration of MNs, the emergence of the subtype identities of MNs, and the projection of motor axons. These findings show that HB9 has an essential function in consolidating the identity of postmitotic MNs. PMID- 10482235 TI - Active suppression of interneuron programs within developing motor neurons revealed by analysis of homeodomain factor HB9. AB - Sonic hedgehog (Shh) specifies the identity of both motor neurons (MNs) and interneurons with morphogen-like activity. Here, we present evidence that the homeodomain factor HB9 is critical for distinguishing MN and interneuron identity in the mouse. Presumptive MN progenitors and postmitotic MNs express HB9, whereas interneurons never express this factor. This pattern resembles a composite of the avian homologs MNR2 and HB9. In mice lacking Hb9, the genetic profile of MNs is significantly altered, particularly by upregulation of Chx10, a gene normally restricted to a class of ventral interneurons. This aberrant gene expression is accompanied by topological disorganization of motor columns, loss of the phrenic and abducens nerves, and intercostal nerve pathfinding defects. Thus, MNs actively suppress interneuron genetic programs to establish their identity. PMID- 10482236 TI - Convergent inductive signals specify midbrain, hindbrain, and spinal cord identity in gastrula stage chick embryos. AB - In the chick embryo, neural cells acquire midbrain, hindbrain, and spinal cord character over a approximately 6 hr period during gastrulation. The convergent actions of four signals appear to specify caudal neural character. Fibroblast growth factors (FGFs) and a paraxial mesoderm-caudalizing (PMC) activity are involved, but neither signal is sufficient to induce any single region. FGFs act indirectly by inducing mesoderm that expresses PMC and retinoid activity and also directly on prospective neural cells, in combination with PMC activity and a rostralizing signal, to induce midbrain character. Hindbrain character emerges from cells that possess the potential to acquire midbrain character upon exposure to higher levels of PMC activity. Induction of spinal cord character appears to involve PMC and retinoid activities. PMID- 10482237 TI - Chemorepulsion of neuronal migration by Slit2 in the developing mammalian forebrain. AB - Newborn cerebral cortical neurons migrate along radial glia to the cortical plate. Experiments using a collagen gel assay revealed that the choroid plexus repelled cerebral cortical neurons and olfactory interneuron precursors, which were mimicked by Neuro-2A cells. Fractionation of Neuro-2A-conditioned medium identified a protein of 190 kDa, equivalent to full-length Slit proteins. Indeed, it cross-reacted with an antibody against Slit2, suggesting that it is either Slit2 or another Slit protein. Further, Slit2, expressed in COS cells, repelled cerebral cortical neurons and olfactory interneuron precursors. Thus, Slit2, which is expressed by the choroid plexus and the septum, acts as a chemorepulsive factor for neuronal migration. These results suggest chemorepulsion as a guidance mechanism for neuronal migration in the developing forebrain. PMID- 10482238 TI - Schwann cell-derived Desert hedgehog controls the development of peripheral nerve sheaths. AB - We show that Schwann cell-derived Desert hedgehog (Dhh) signals the formation of the connective tissue sheath around peripheral nerves. mRNAs for dhh and its receptor patched (ptc) are expressed in Schwann cells and perineural mesenchyme, respectively. In dhh-/- mice, epineurial collagen is reduced, while the perineurium is thin and disorganized, has patchy basal lamina, and fails to express connexin 43. Perineurial tight junctions are abnormal and allow the passage of proteins and neutrophils. In nerve fibroblasts, Dhh upregulates ptc and hedgehog-interacting protein (hip). These experiments reveal a novel developmental signaling pathway between glia and mesenchymal connective tissue and demonstrate its molecular identity in peripheral nerve. They also show that Schwann cell-derived signals can act as important regulators of nerve development. PMID- 10482239 TI - GFR alpha3, a component of the artemin receptor, is required for migration and survival of the superior cervical ganglion. AB - GFR alpha3 is a component of the receptor for the neurotrophic factor artemin. The role of GFR alpha3 in nervous system development was examined by generating mice in which the Gfr alpha3 gene was disrupted. The Gfr alpha3-/- mice exhibited severe defects in the superior cervical ganglion (SCG), whereas other ganglia appeared normal. SCG precursor cells in the mutant embryos failed to migrate to the correct position, and they subsequently failed to innervate the target organs. In wild-type embryos, Gfr alpha3 was expressed in migrating SCG precursors, and artemin was expressed in and near the SCG. After birth, SCG neurons in the mutant mice underwent progressive cell death. These observations suggest that GFR alpha3-mediated signaling is required both for the rostral migration of SCG precursors and for the survival of mature SCG neurons. PMID- 10482240 TI - Functional dissociation of mu opioid receptor signaling and endocytosis: implications for the biology of opiate tolerance and addiction. AB - Opiate analgesia, tolerance, and addiction are mediated by drug-induced activation of the mu opioid receptor. A fundamental question in addiction biology is why exogenous opiate drugs have a high liability for inducing tolerance and addiction while native ligands do not. Studies indicate that highly addictive opiate drugs such as morphine are deficient in their ability to induce the desensitization and endocytosis of receptors. Here, we demonstrate that this regulatory mechanism reveals an independent functional property of opiate drugs that can be distinguished from previously established agonist properties. Moreover, this property correlates with agonist propensity to promote physiological tolerance, suggesting a fundamental revision of our understanding of the role of receptor endocytosis in the biology of opiate drug action and addiction. PMID- 10482241 TI - The generality of parietal involvement in visual attention. AB - Functional magnetic resonance imaging (fMRI) was used to determine whether different kinds of visual attention rely on a common neural substrate. Within one session, subjects performed three different attention experiments (each comparing an attentionally demanding task with an easier task using identical stimuli): (1) peripheral shifting, (2) object matching, and (3) a nonspatial conjunction task. Two areas were activated in all three experiments: one at the junction of intraparietal and transverse occipital sulci (IPTO), and another in the anterior intraparietal sulcus (AIPS). These regions are not simply involved in any effortful task, because they were not activated in a fourth experiment comparing a difficult language task with an easier control task. Thus, activity in IPTO and AIPS generalizes across a wide variety of attention-requiring tasks, supporting the existence of a common neural substrate underlying multiple modes of visual selection. PMID- 10482242 TI - Effects of attention on the reliability of individual neurons in monkey visual cortex. AB - To determine the physiological mechanisms underlying the enhancement of performance by attention, we examined how attention affects the ability of isolated neurons to discriminate orientation by investigating the reliability of responses with and without attention. Recording from 262 neurons in cortical area V4 while two rhesus macaques did a delayed match-to-sample task with oriented stimuli, we found that attention did not produce detectable changes in the variability of neuronal responses but did improve the orientation discriminability of the neurons. We also found that attention did not change the relationship between burst rate and response rate. Our results are consistent with the idea that attention selects groups of neurons for a multiplicative enhancement in response strength. PMID- 10482243 TI - Leptin differentially regulates NPY and POMC neurons projecting to the lateral hypothalamic area. AB - Recent studies have reinforced the view that the lateral hypothalamic area (LHA) regulates food intake and body weight. We identified leptin-sensitive neurons in the arcuate nucleus of the hypothalamus (Arc) that innervate the LHA using retrograde tracing with leptin administration. We found that retrogradely labeled cells in the Arc contained neuropeptide Y (NPY) mRNA or proopiomelanocortin (POMC) mRNA. Following leptin administration, NPY cells in the Arc did not express Fos but expressed suppressor of cytokine signaling-3 (SOCS-3) mRNA. In contrast, leptin induced both Fos and SOCS-3 expression in POMC neurons, many of which also innervated the LHA. These findings suggest that leptin directly and differentially engages NPY and POMC neurons that project to the LHA, linking circulating leptin and neurons that regulate feeding behavior and body weight homeostasis. PMID- 10482244 TI - Calcium-stimulated adenylyl cyclase activity is critical for hippocampus dependent long-term memory and late phase LTP. AB - It is hypothesized that Ca2+ stimulation of calmodulin (CaM)-activated adenylyl cyclases (AC1 or AC8) generates cAMP signals critical for late phase LTP (L-LTP) and long-term memory (LTM). However, mice lacking either AC1 or AC8 exhibit normal L-LTP and LTM. Here, we report that mice lacking both enzymes (DKO) do not exhibit L-LTP or LTM. To determine if these defects are due to a loss of cAMP increases in the hippocampus, DKO mice were unilaterally cannulated to deliver forskolin. Administration of forskolin to area CA1 before training restored normal LTM. We conclude that Ca2+-stimulated adenylyl cyclase activity is essential for L-LTP and LTM and that AC1 or AC8 can produce the necessary cAMP signal. PMID- 10482245 TI - Calmodulin mediates calcium-dependent inactivation of the cerebellar type 1 inositol 1,4,5-trisphosphate receptor. AB - The dependency of purified mouse cerebellar type 1 inositol 1,4,5-trisphosphate receptor (IP3R1)/Ca2+ channel function on cytoplasmic Ca2+ was examined. In contrast to the channels in crude systems, the purified IP3R1 reconstituted into planar lipid bilayers did not show the bell-shaped dependence on Ca2+. It was activated with increasing Ca2+ sublinearly without inhibition even up to 200 microM. The addition of calmodulin to the cytoplasmic side inhibited the channel at high Ca2+ concentrations. Calmodulin antagonists reversed the Ca2+-dependent inactivation of the native channels in cerebellar microsomes. These results indicate that the bell-shaped dependence on cytoplasmic Ca2+ is not an intrinsic property of the IP3R1, and the Ca2+-dependent inactivation is directly mediated by calmodulin. PMID- 10482246 TI - Stoichiometry and arrangement of subunits in rod cyclic nucleotide-gated channels. AB - Cyclic nucleotide-gated (CNG) ion channels mediate the response to light in retinal rods. They are tetramers of two homologous subunits (alpha and beta), each of which is essential for the function of the channels in vivo. We have investigated the stoichiometry and arrangement of these two subunits to determine how they come together within an individual channel complex. We exploited the very specific geometric and spatial requirements for forming a high-affinity Ni2+ binding site to examine the number and relative positions of the subunits. We found that only an order of alpha/alpha/beta/beta could account qualitatively and quantitatively for the observed intersubunit coordination of Ni2+ in wild-type and mutant alpha/beta channels. Furthermore, our results suggest a structural dimerization among like subunits, at least at the level of the Ni2+-binding site. PMID- 10482247 TI - The reduced release probability of releasable vesicles during recovery from short term synaptic depression. AB - Recovery from synaptic depression is believed to depend mainly on replenishment of the releasable pool of vesicles. We observed that during recovery from depression in a calyx-type synapse, part of the releasable pool was replenished rapidly. Half recovery occurred within 1 s, even in the absence of residual calcium. Vesicles that had recently entered the releasable pool had a 7- to 8 fold lower release probability than those that had been in the pool for more than 30 s. These results suggest that the reduction in the release probability of releasable vesicles contributes greatly to the level of depression. How synapses maintain transmission during repetitive firing is in debate. We propose that during repetitive firing, accumulation of intracellular Ca2+ may facilitate release of the rapidly replenished but reluctant vesicles, making them available for sustaining synaptic transmission. PMID- 10482248 TI - Immunocytochemical localisation of PTHrP (parathormone-related peptide) in myoepithelial cells of human sweat and parotid glands. AB - PTHrP is a HHM-inducing peptide. It exhibits certain structural similarity to PTH and the two hormones may act through the same receptors. PTHrP is known to be produced in various tissues as well as during development. In this study we decided to immunocytochemically demonstrate PTHrP in normal skin and squamous cell carcinomas as well as in parotid glands (normal, inflamed and neoplastic). In the skin, PTHrP expression was demonstrated in epidermis and in smooth muscle cell layer of blood vessels. In squamous cell carcinomas, the expression was noted in foci of keratinization. In parotid glands, the peptide was localised in excretory ducts and in blood vessels, while inflammation of the gland and its tumours resulted most frequently in the less intense immunoreaction. The results are consistent with those of other authors. The novel observations include demonstration of PTHrP expression in myoepithelial cells of sweat glands and in parotid glands, where it may be involved in the control of their contractile activity. PMID- 10482249 TI - Diversity of the plasma membrane properties of transplantable hamster melanomas with regard to the expression of P-glycoprotein. AB - A comparison of the expression of P-glycoprotein (Pgp) was performed in two forms of hamster transplantable melanomas of common origin, but differing in growth rates and levels of differentiation. The expression of P-glycoprotein in plasma membranes of these two forms of melanomas was estimated by the western blot analysis and the transport activity of the Pgp compared by flow cytometry. It was observed that a spontaneous alteration in the original melanotic melanoma leading to a formation of the amelanotic form characterized by higher growth rate, greater anaplasticity and leading to the animals' death after a shorter time from inoculation, was accompanied by a decrease in the Pgp expression and activity, due to simultaneous appearance of a small population of amelanotic cells with high Pgp expression and activity, and disappearance of this activity from the major population. It is possible, that the activity of Pgp in the melanoma cell membranes reflects the degree of cell differentiation. PMID- 10482250 TI - Extrachromosomal rDNA and polarity of pro-oocytes during ovary development in Creophilus maxillosus (Coleoptera, Staphylinidae). AB - In telotrophic ovary of Creophilus maxillosus, the differentiation of the oocyte and nurse cells takes place within the linear clusters of sister oogonial cells. The amplification of rDNA occurs in the nuclei of pro-oocytes which are the most posterior cells of the clusters. During the consecutive oogonial divisions extrachromosomal rDNA segregates preferentially to the pro-oocyte of the next generation. We analyzed the ultrastructure of pro-oocytes and pro-nurse cells in the early and late phase of rDNA amplification in pupal ovary of Creophilus maxillosus. We found that pro-oocytes of the same generation contain variable amounts of extrachromosomal rDNA and that the presence of extra DNA is not limited to the nuclei of pro-oocytes; extra DNA is also present in the nuclei of some pro-nurse cells. Pro-oocytes can experience partial loss of extrachromosomal DNA during early oogonial divisions which is caused by the imprecise segregation of this material to the posterior pole. We believe that this imperfect segregation is a source of extrachromosomal DNA present in the nuclei of pro nurse cells. Ultrastructural analysis showed that multiple nucleoli do not disperse in oogonial mitoses but remain associated with extrachromosomal chromatin and segregate with it to the posterior pole of the pro-oocyte. We also analyzed the ultrastructure of the germ plasm--a cytoplasmic structure present at the posterior pole of pro-oocytes. We have found that this structure contains spectrin and at the ultrastructural level is strikingly similar to the spectrosome which is present in germline cells of Drosophila. We also found spectrin in the intercellular bridges which connect oogonial cells and are known to contain fusomes. PMID- 10482251 TI - Holoblastic early cleavage of Tetrodontophora bielanensis (Collembola) eggs, with special reference to its irregularity. AB - The fertilized eggs of Tetrodontophora bielanensis start to cleave 6 to 8 days after oviposition and initially only karyokineses occur. The cytokinesis begins after two karyokineses, when four nuclei are observed in the ooplasm. Two cleavage furrows, perpendicular to each other, appear simultaneously at the egg poles where polar bodies are located and gradually the furrows encompass the whole egg diameter. The furrow formation is initiated by the bundle of microfilaments that contract and pull superficial fragments of the oolemma into the yolk and subsequently new membranes, separating the daughter cells, start to form. However, they do not grow towards the egg centre but bifurcate, leaving the central part of the ooplasm outside of the newly formed blastomeres. Starting from the fourth or fifth cleavage division, the bifurcations permanently occur and multiple cleavage furrows are formed on the embryo surface. Moreover, fragments of the ooplasm, enclosed within the cell membrane but devoid of cell nucleus are observed. During further development such cell fragments become reincorporated into the embryo. This mode of cleavage leads eventually to the formation of cellular blastoderm on the embryo surface. The results presented in the paper suggest that the control of cleavage in T. bielanensis acts not at the level of cytoplasmic determinants but rather at the level of positional information of blastomeres. PMID- 10482252 TI - Immunocytochemical localization of pectins in the maturing anther of Allium cepa L. AB - Distribution of pectins in cell walls of maturing anther of Allium cepa L. was investigated. The monoclonal antibodies against defined epitopes of pectin were used: JIM5 recognizing unesterified pectin and JIM7 recognizing esterified pectin. It has been found that the cell walls of all anther tissues mainly contain esterified pectins. In the somatic tissues only small amounts of unesterified pectins are present in the cell wall junctions and adjacent middle lamellae and in the cell walls of the connective tissue. Thickening of the epiderm cell walls and growth of trabeculae in endothecium are completed through deposition of esterified pectins. In the cell walls of the middle layer and tapetum, unesterified pectins have been found only prior to their disintegration. The primary wall of microsporocytes is made up mainly of esterified pectins. Unesterified pectins occur outside microsporocytes only prior to the callose isolation stage. The presence of esterified pectins has also been detected on the surface of the callose wall surrounding dividing microsporocytes. Lysis of those pectins takes place after microsporogenesis, simultaneously with the lysis of the callosic walls. Before these processes pectins are unesterified. In the sporoderm of pollen grains mainly esterified pectins occur. They have been localized in the intine and aperture. The level of unesterified pectins in the intine is markedly lower. PMID- 10482253 TI - Effects of antheridial peptides derived from Chara on cell cycle-regulated processes in human and yeast cells. AB - Previous experiments have shown that extracts obtained from maturing male sex organs of Chara tomentosa, containing a low molecular weight peptide (termed antheridial chromatin condensation factor--ACCF) are capable to induce structural and functional effects in root meristems and fern gametophytes. Our present data point to a number of resemblances between the phenotypic characters of antheridial filaments (a.f.) and the properties of ACCF-treated human lymphocytes and HeLa cells; these comprise primarily a number of morphological changes at the nuclear/chromosomal levels. Mitotic chromosomes become shortened and the relative duration of prophase is reduced, while duration of telophase is prolonged. Nucleolar profiles in ACCF-treated HeLa cells become reduced. Significant decrease in mitotic activity was found in human and yeast cells (Schizosaccharomyces pombe). All the above similarities between the "innate" processes within a.f. and those induced by ACCF provide positive evidence for the presence of a highly specific factor that contributes to nuclear re-patterning of cells undergoing morphogenetic transformations before the onset of spermiogenesis. PMID- 10482254 TI - Disappearance of 3H-leucine incorporation circadian rhythm induced by symplasmic isolation of Chara vulgaris antheridium from thallus. AB - Autoradiographic research with the use of 3H-leucine demonstrates that circadian rhythm of protein synthesis characteristic of manubria at the proliferative phase of spermatogenesis in Chara vulgaris disappears after symplasmic isolation of antheridium from thallus during the time preceding block of DNA endoreplication in manubria following initiation of spermiogenesis insensitive to light. PMID- 10482255 TI - Ketanserin, an antagonist of 5-HT2A receptor of serotonin, inhibits testosterone secretion by rat Leydig cells in vitro. AB - The effect of ketanserin, an antagonist of 5-HT2A receptor of serotonin, added to the culture medium, on basal and LH-stimulated testosterone secretion was studied in primary cultures of adolescent rat Leydig cells. Ketanserin decreased the basal secretion of testosterone but showed an insignificant influence on the LH stimulated process. It can be concluded that ketanserin may affect the testosterone-secreting cells by an indirect action at the vascular level as well as directly at the level of Leydig cells, at least in adolescent rats, leading to down-regulation of the basal testosterone secretion. PMID- 10482256 TI - The evolution of swallowing neuroanatomy and physiology in humans: a practical perspective. PMID- 10482257 TI - The functional neuroanatomy of voluntary swallowing. AB - Swallowing is a complex physiological process involving voluntary and reflexive motor activity, sensorimotor integration, salivation, and visceral regulation. Despite the numerous processes required for normal deglutition, traditional models of the central control of swallowing only emphasize the involvement of the brainstem and the inferior precentral gyrus (IPCG). However a number of neurological disorders involving other brain regions also cause dysphagia. To determine the brain regions participating in voluntary swallowing, we assayed regional cerebral blood flow (rCBF) with positron emission tomography (PET) while healthy human subjects swallowed, performed lateral tongue movements, or rested with their eyes closed. Voluntary swallowing produced strong rCBF increases within the IPCG bilaterally, the right anterior insula/claustrum, and the left cerebellum. The maxima in these regions differed from those induced by lateral tongue movements. Swallowing also produced rCBF increases in the putamen, thalamus, and several additional cortical areas, but these foci were not as clearly distinguishable from activity arising during tongue movements. These findings indicate that swallowing involves the recruitment of a large-scale distributed neural network that includes the anterior insula and cerebellum. The distributed nature of this network helps to explain why so many neurological conditions produce dysphagia. PMID- 10482258 TI - Ocular motor abnormalities in ataxia telangiectasia. AB - Although abnormal eye movements are a prominent feature of ataxia telangiectasia, the characteristics of the oculomotor dysfunction in the disease have been reported only in small groups of patients. We have examined eye movements clinically in 56 patients with ataxia telangiectasia, and obtained electrooculographic recordings of eye movements in 33 subjects. Deficits were observed in the eye movement systems that stabilize images on the retina, including pursuit, gaze holding, convergence, vestibular and optokinetic slow phases, and cancellation of vestibular slow phases. Abnormalities in the systems that maintain fixation and shift gaze were also prominent, including abnormal reflexive and voluntary saccades (characterized by prolonged latency, hypometric amplitude, and the use of head movements to initiate gaze shifts), impaired fixation, and a reduction in vestibular and optokinetic quick phases. The abnormalities in image stabilization most likely result from dysfunction in the cerebellar flocculus and paraflocculus. The basis of the saccadic and fixation disturbance is less certain but may be the result of abnormal supranuclear control of the superior colliculus resulting from dysfunction in the cerebellar vermis or the basal ganglia. PMID- 10482259 TI - Monoclonal antibody treatment exposes three mechanisms underlying the clinical course of multiple sclerosis. AB - The elective treatment of patients with multiple sclerosis, using a humanized anti-leukocyte (CD52) monoclonal antibody (Campath-1H), has illuminated mechanisms that underlie the clinical course of the disease. Twenty-seven patients were studied clinically and by magnetic resonance imaging (MRI) before and for 18 months after a single pulse of Campath-1H. The first dose of monoclonal antibody was associated with a transient rehearsal of previous symptoms caused by the release of mediators that impede conduction at previously demyelinated sites; this effect remained despite selective blockade of tumor necrosis factor-alpha. Disease activity persisted for several weeks after treatment but thereafter radiological markers of cerebral inflammation were suppressed for at least 18 months during which there were no new symptoms or signs. However, about half the patients experienced progressive disability and increasing brain atrophy, attributable on the basis of MRI spectroscopy to axonal degeneration, which correlated with the extent of cerebral inflammation in the pretreatment phase. These data support the formulation that inflammation and demyelination are responsible for relapses of multiple sclerosis; that inflammatory mediators, but not tumor necrosis factor-alpha, cause symptomatic reactivation of previously demyelinated lesions; and that axonal degeneration, conditioned by prior inflammation but proceeding despite its suppression, contributes to the progressive phase of disability. These results provide evidence supporting the emerging view that treatment in multiple sclerosis must be given early in the course, before the consequences of inflammation are irretrievably established. PMID- 10482260 TI - A reduced K+ current due to a novel mutation in KCNQ2 causes neonatal convulsions. AB - Benign familial neonatal convulsions (BFNC) is a rare dominantly inherited epileptic syndrome characterized by frequent brief seizures within the first days of life. The disease is caused by mutations in one of two recently identified voltage-gated potassium channel genes, KCNQ2 or KCNQ3. Here, we describe a four generation BFNC family carrying a novel mutation within the distal, unconserved C terminal domain of KCNQ2, a 1-bp deletion, 2513delG, in codon 838 predicting substitution of the last seven and extension by another 56 amino acids. Three family members suffering from febrile but not from neonatal convulsions do not carry the mutation, confirming that febrile convulsions and BFNC are of different pathogenesis. Functional expression of the mutant channel in Xenopus oocytes revealed a reduction of the potassium current to 5% of the wild-type current, but the voltage sensitivity and kinetics were not significantly changed. To find out whether the loss of the last seven amino acids or the C-terminal extension because of 2513delG causes the phenotype, a second, artificial mutation was constructed yielding a stop codon at position 838. This truncation increased the potassium current by twofold compared with the wild type, indicating that the pathological extension produces the phenotype, and suggesting an important role of the distal, unconserved C-terminal domain of this channel. Our results indicate that BFNC is caused by a decreased potassium current impairing repolarization of the neuronal cell membrane, which results in hyperexcitability of the central nervous system. PMID- 10482261 TI - Myelin deficiencies in both the central and the peripheral nervous systems associated with a SOX10 mutation. AB - We describe an unique patient presenting with severe leukodystrophy compatible with Pelizaeus-Merzbacher disease and peripheral neuropathy consistent with Charcot-Marie-Tooth disease type 1 in addition to Waardenburg-Hirschsprung syndrome. A novel mutation was identified in her SOX10 gene, which encodes a transcription factor preferentially expressed in the late embryonic glial cell lineage and in mature myelin-forming cells of both the central nervous system and peripheral nervous system, as well as in the early neural crest cells. Heterozygous SOX10 loss-of-function mutations have been reported in patients with Waardenburg-Hirschsprung syndrome and its murine model, Dominant megacolon. However, neither Waardenburg-Hirschsprung syndrome patients nor Dominant megacolon mice have dysmyelinating features, suggesting the question of how SOX10 acts in the glial lineage in vivo. The novel mutation described herein does not disrupt the coding region but extends the peptide and hence is likely to act as a dominant-negative allele. Our findings indicate that dysfunction of SOX10 may lead to deficiency of myelination in the central nervous system and peripheral nervous system as well as hypopigmentation and enteric aganglionosis. PMID- 10482262 TI - Migratory behavior of lymphocytes isolated from multiple sclerosis patients: effects of interferon beta-1b therapy. AB - Previous reports by our group and by others have shown that in vitro treatment of T cells derived from healthy, normal subjects with interferon beta-1b (IFN beta1b) reduces metalloproteinase (metalloproteinase type 9 [MMP-9]) activity with a consequent reduction in lymphocyte migration. In this study, we used a Boyden chamber assay to assess the migratory capacity of T cells derived from multiple sclerosis patients who either did or did not receive IFN-beta1b. Lymphocytes derived from patients treated for less than 2 years with IFN-beta migrated at a low rate that was indistinguishable from that of cells isolated from healthy donors. However, longer term treatment with IFN (>3.5 years) was associated with a reversion of the migration to a high level that did not differ statistically from that of cells isolated from untreated multiple sclerosis patients. For both high-migratory groups, migration could be reduced to control levels after the exogenous addition of TIMP-1, a relatively specific inhibitor of the MMP-9, implicating this protease in the process of T-cell migration. Our findings suggest that one of the mechanisms by which IFN-beta exerts its action is by reducing MMP-9 activity and thus the entry of inflammatory cells into the nervous system and, as such, MMPs may constitute potential therapeutic targets in inflammatory diseases such as multiple sclerosis. PMID- 10482263 TI - Overexpression of four-repeat tau mRNA isoforms in progressive supranuclear palsy but not in Alzheimer's disease. AB - Perturbations in the microtubule-associated protein tau occur in several human neurodegenerative diseases. In Alzheimer's disease and progressive supranuclear palsy (PSP), tau proteins assemble into straight and paired helical filaments that form intraneuronal deposits of neurofibrillary tangles (NFTs). The mechanisms underlying the aberrant assembly of tau into NFTs is unknown. To determine whether alterations in the expression of the carboxyl-terminal variants of tau contribute to NFT formation, we analyzed tau mRNA isoform expression in select regions of control, Alzheimer's disease, and PSP brains. In Alzheimer's disease, there were no alterations in tau mRNA isoform expression. However, in PSP, the levels of tau mRNA isoforms containing four microtubule binding domains were increased in the brainstem but not the frontal cortex or cerebellum. The brainstem in PSP has extensive NFT pathology, whereas the frontal cortex and cerebellum are relatively spared, suggesting that alterations in tau mRNA isoform expression occur in NFT-vulnerable regions in this disease. An increase in the four-repeat tau mRNA may lead to an increase in four-repeat tau protein isoforms and may contribute to the formation of NFTs in PSP. A similar increase in four repeat tau mRNA has been reported for mutations associated with frontotemporal dementia and parkinsonism linked to chromosome 17. PMID- 10482264 TI - Reversal of acute apparent diffusion coefficient abnormalities and delayed neuronal death following transient focal cerebral ischemia in rats. AB - Twenty-two rats were subjected to 8, 15, 30, or 60 minutes of temporary middle cerebral artery occlusion (n = 5 per group) or sham occlusion (n = 2) in the magnetic resonance imaging unit. Diffusion-, perfusion-, and T2-weighted imaging were acquired before and during occlusion, and after reperfusion. A coregistration method was used to correlate the acute changes of the average apparent diffusion coefficient (ADCav) with the histology after 72 hours at the same topographic sites. The initially reduced ADCav values recovered completely in both the lateral caudoputamen and upper frontoparietal cortex in the 8-, 15-, and 30-minute groups, partially in the cortex, and not at all in the caudoputamen in the 60-minute group. The histology showed that the caudoputamen was either normal or had mild neuronal injury in the 8-minute group and invariably had some degree of neuronal death in the 15-, 30-, and 60-minute groups, whereas the cortex was either normal or had varying degrees of neuronal injury in all groups. No histological abnormalities were seen in the sham-operated rats. Our data suggest that acute ADCav reversal does not always predict tissue recovery from ischemic injury and that temporary focal ischemia for even 8-minute duration can cause delayed neuronal death that is more severe in the caudoputamen where the initial ADCav decline was greater than in the cortex. PMID- 10482265 TI - Hippocampal N-methyl-D-aspartate receptor subunit mRNA levels in temporal lobe epilepsy patients. AB - Changes in the subunit stoichiometry of the N-methyl-D-aspartate (NMDA) receptor (NMDAR) alters its channel properties, and may enhance or reduce neuronal excitability in temporal lobe epilepsy patients. This study determined whether hippocampal NMDA receptor subunit mRNA levels were increased or decreased in temporal lobe epilepsy patients compared with nonseizure autopsy cases. Hippocampal sclerosis (HS; n = 16), non-HS (n = 10), and autopsy hippocampi (n = 9) were studied for NMDAR1 (NR1) and NR2A-D mRNA levels by using semiquantitative in situ hybridization techniques, along with neuron densities. Compared with autopsy hippocampi, non-HS and HS patients showed increased NR2A and NR2B hybridization densities per dentate granule cell. Furthermore, non-HS hippocampi showed increased NR1 and NR2B mRNA levels per CA2/3 pyramidal neuron compared with autopsy cases. HS patients, by contrast, showed decreased NR2A hybridization densities per CA2/3 pyramidal neuron compared with non-HS and autopsy cases. These findings indicate that chronic temporal lobe seizures are associated with differential changes in hippocampal NR1 and NR2A-D hybridization densities that vary by subfield and clinical-pathological category. In temporal lobe epilepsy patients, these findings support the hypothesis that in dentate granule cells NMDA receptors are increased, and excitatory postsynaptic potentials should be strongly NMDA mediated compared with nonseizure autopsies. HS patients, by comparison, showed decreased pyramidal neuron NR2A mRNA levels, and this suggests that NMDA-mediated pyramidal neuron responses should be reduced in HS patients compared with non-HS cases. PMID- 10482266 TI - Regulation of dopaminergic activity in early Parkinson's disease. AB - Parkinson's disease (PD) is characterized by an uneven and progressive loss of nigrostriatal dopaminergic neurons. It is hypothesized that the physiological basis for the therapeutic response in early stages of PD is the ability for the partially and unevenly denervated dopaminergic system to restore and normalize dopaminergic influence in functionally segregated subregions of the basal ganglia. To investigate this hypothesis, patients with early and uncomplicated PD were investigated with positron emission tomography by using a two-tracer protocol yielding a measure of dopamine transporter-corrected dopamine synthesis capacity. Compared with controls, patients with PD exhibited a considerable increase in dopamine transporter-corrected dopamine synthesis capacity. The increase showed an inverse dependence on the structural integrity in as much as the highest rate was measured in the most denervated region, the dorsal part of putamen (198% of control value). A therapeutic challenge with antiparkinsonian medication state-dependently decreased dopaminergic activity. Thus, it is demonstrated that dopaminergic degeneration in PD is accompanied by a conspicuous acceleration of presynaptic dopaminergic activity, which is state-dependently down-regulated by dopaminomimetic treatment. It is suggested that homeostatic mechanisms acting to maintain congruity within the dopaminergic system are functionally intact in early PD. PMID- 10482267 TI - Sequence-specific cleavage of Huntingtin mRNA by catalytic DNA. AB - The selective loss of neurons in Huntington's disease (HD) is caused by the abnormal expansion of the CAG triplet (>36 repeats) of the HD gene. Although the molecular events that lead to neuronal death are not clear, it is most likely that mutant HD protein operates through a "gain-of-function" mechanism. One possible therapeutic approach that does not require definition of the toxic mechanism(s) involves reduction in the levels of mutant HD protein by decreasing the quantity of translatable HD mRNA. In this report, we demonstrate the first effective destruction of the HD mRNA, using a catalytic DNA--an oligodeoxynucleotide with RNA-cleaving enzymatic activity. We show that the cleavage of HD mRNA by the catalytic DNA occurs in a sequence-specific manner, and leads to significant reduction of HD protein expression in mammalian cells. The catalytic DNAs we have developed are a valuable research tool for studying HD, and may have the therapeutic potential of reducing cellular toxicity caused by mutant HD protein. PMID- 10482268 TI - Reduced expression of the G209A alpha-synuclein allele in familial Parkinsonism. AB - Missense mutations at the alpha-synuclein gene have been associated with familial parkinsonism. We report that the phenotype of a kindred (Family H) with autosomal dominant, levodopa-responsive parkinsonism maps to chromosomal region 4q21-23 and that affected members of this kindred harbor a previously reported mutation (G209A) in exon 4 of the alpha-synuclein gene. We assessed the expression of the G209A allele in lymphoblastoid cell lines established from 12 individuals heterozygous for the G209A allele. The expression of this allele is either absent or significantly reduced in 7 affected heterozygotes and in 3 asymptomatic heterozygotes who are older than the mean age at disease diagnosis for their generation. In contrast, it is expressed in 1 affected and 1 unaffected heterozygote. The unaffected heterozygote is younger than the mean age at disease diagnosis for their generation. The lack of or significantly reduced expression of the G209A allele in affected heterozygotes suggests that the timing of reduced expression may be critical for disease onset. If so, the parkinsonian phenotype may arise from haploinsufficiency at the alpha-synuclein gene at a time point before symptom onset. In conclusion, reduced alpha-synuclein gene expression may be important in the pathogenesis of parkinsonism. PMID- 10482269 TI - Genetic association of alpha2-macroglobulin with Alzheimer's disease in a Finnish elderly population. AB - Recently, two studies have reported an association between the alpha2 macroglobulin gene on chromosome 12 and late-onset Alzheimer's disease, whereas others have not been able to replicate these findings. By using a prospective population-based study, we have investigated the relation between two polymorphisms in this gene with the presence of the disease and also with the extent of pathological changes in the cerebral cortex. The Vantaa 85+ Study includes all 601 persons, at least 85 years of age, who were living in Vantaa, Finland, on April 1, 1991. The neocortical beta-amyloid protein load and the number of neurofibrillary tangles were determined on tissue sections by using methenamine silver staining and a modified Bielschowsky staining, respectively. The A/A genotype in exon 24 of the alpha2-macroglobulin gene was associated with neuropathologically defined diagnosis of Alzheimer's disease according to the CERAD (Consortium to Establish a Registry for Alzheimer's Disease) criteria and with an increase in the neocortical beta-amyloid protein load. The effect of this association was stronger in the apolipoprotein E epsilon4-negative group. Therefore, genetic variability in the alpha2-macroglobulin gene is a risk factor associated with neuropathologically defined Alzheimer's disease in our population, as well as with the extent of neocortical beta-amyloid protein deposition. PMID- 10482271 TI - Genetic linkage of Welander distal myopathy to chromosome 2p13. AB - Welander distal myopathy (WDM) is an autosomal dominant myopathy with late-adult onset characterized by slow progression of distal muscle weakness. The disorder is considered a model disease for hereditary distal myopathies and is almost only seen in Sweden and some parts of Finland. A genomewide screening has been performed in initially two Swedish families with 400 highly polymorphic microsatellite markers. We report here that the disease is linked to chromosome 2p13. Seven additional nonrelated families have subsequently been mapped to the same area where a maximum two-point LOD score of 17.97 was obtained with the marker D2S2113 at 0.0 recombination fraction. The region has been restricted by recombinations and the finding of a common shared haplotype through all analyzed families. This restricts the gene locus region to 2.4 cM. These findings provide evidence for the involvement of a single locus for WDM. The WDM region overlaps with the linkage region for Miyoshi myopathy and limb-girdle muscular dystrophy 2B. The dysferlin gene responsible for these disorders is considered a primary candidate gene for WDM. PMID- 10482270 TI - Cerebrospinal fluid levels of MMP-2, 7, and 9 are elevated in association with human immunodeficiency virus dementia. AB - Pathological evidence suggests that alterations of the blood-brain barrier (BBB) may occur in association with human immunodeficiency virus (HIV) dementia (HIVD). Increased BBB permeability could contribute to the development of dementia by facilitating the entry of activated and infected monocytes, as well as potentially toxic serum proteins, into the central nervous system. One mechanism by which BBB permeability may be altered is through increased activity of select matrix metalloproteinases (MMPs). In the present study, we examined the possibility that MMPs that target critical BBB proteins, including laminin, entactin, and collagen type IV, are elevated in the cerebrospinal fluid (CSF) of patients with HIVD. We also examined the possibility that such MMPs could be produced by brain-derived cells, and that MMP production by these cells might be increased by tumor necrosis factor-alpha, an inflammatory cytokine that is produced by HIV-infected monocytes/microglia and is elevated in HIVD. By using western blot and enzyme-linked immunosorbent assay, we observed that CSF levels of pro-MMP-2 and pro-MMP-7 were increased in association with HIVD. In addition, through the use of gelatin substrate zymography, a sensitive functional assay for MMP-2 and MMP-9, we observed that MMP-2 or pro-MMP-9 activity was more frequently detectable in the CSF of individuals with HIV dementia (9/16) than in the CSF from either nondemented seropositive (2/11) or seronegative (0/11) controls. Although the presence of MMPs in the serum could contribute to elevated levels in the CSF, we also show that brain-derived cells release MMP-2, 7, and 9, and that such release is increased after their stimulation with tumor necrosis factor alpha. Together, these results suggest that elevated CSF levels of select MMPs may reflect immune activation within the central nervous system. They also suggest that further studies may be warranted to determine whether these proteins may play a role in the development of symptomatic neurological disease. PMID- 10482272 TI - Language area localization with three-dimensional functional magnetic resonance imaging matches intrasulcal electrostimulation in Broca's area. AB - In this study, intraoperative electrocortical stimulation mapping (ioESM), the current gold standard for the localization of critical language areas, is compared with functional magnetic resonance imaging (fMRI) in a 14-year-old girl with medically intractable epilepsy caused by a tumor in the region of Broca's area. Prior to the operation, four different fMRI tasks that target inferior frontal language areas were applied. Prior to the resection, ioESM as well as fMRI detected no language areas at the exposed cortical area. After removal of the tumor, a unique opportunity presented itself, where ioESM could be performed in the depth of a now exposed and intact gyrus. One specific locus that was indicated to be a critical language area by multiple-task fMRI was targeted. IoESM selectively confirmed the location of this language area to within an estimated 3 mm. We propose that the combined use of different fMRI tasks increases the sensitivity and specificity for the detection of essential language areas. PMID- 10482273 TI - Role of very-long-chain acyl-coenzyme A synthetase in X-linked adrenoleukodystrophy. AB - X-linked adrenoleukodystrophy (X-ALD) is characterized biochemically by decreased ability of cells to activate (via very-long-chain acyl-coenzyme A synthetase [VLCS]) and subsequently degrade very-long-chain fatty acids in peroxisomes. It is noteworthy that the gene defective in X-ALD encodes ALDP, a peroxisomal membrane protein unrelated to VLCS. We cloned human VLCS (hVLCS) and found that peroxisomes from X-ALD fibroblasts contained immunoreactive hVLCS, refuting the earlier hypothesis that ALDP is required to anchor VLCS to the peroxisomal membrane. Furthermore, hVLCS was topographically oriented facing the peroxisomal matrix in both control and X-ALD fibroblasts, contradicting the alternative hypothesis that ALDP is required to translocate VLCS into peroxisomes. However, overexpression of both hVLCS and ALDP in X-ALD fibroblasts synergistically increased very-long-chain fatty acid beta-oxidation, indicating that these proteins interact functionally. PMID- 10482274 TI - Plasma amyloid beta-peptide 1-42 and incipient Alzheimer's disease. AB - Mutations in the amyloid precursor protein and presenilin 1 and 2 genes result in elevated plasma levels of the amyloid beta-peptide species terminating at amino acid residue 42 (A beta1-42). In a longitudinal study of unrelated elderly individuals, those who subsequently developed Alzheimer's disease had higher plasma levels of A beta1-42 at entry than did those who remained free of dementia. The results indicate that elevated plasma levels of the released A beta peptide A beta1-42 may be detected several years before the onset of symptoms, supporting that extracellular A beta1-42 plays an important role in the pathogenesis of late-onset Alzheimer's disease. PMID- 10482275 TI - Distribution of herpes simplex virus type 1 in human geniculate and vestibular ganglia: implications for vestibular neuritis. AB - Vestibular neuritis is a common cause of partial unilateral vestibular paralysis, which usually spares posterior semicircular canal function. The cause is assumed to be a viral reactivation of latent herpes simplex virus type 1 (HSV-1) in human vestibular ganglia. The existence of an anastomosis between the intermediate nerve and the superior vestibular nerve suggests the question of whether selective affliction of the superior vestibular nerve is the result of migration of HSV-1 from the geniculate ganglion along this faciovestibular anastomosis. We determined the distribution of HSV-1 among geniculate ganglia, vestibular ganglia, and within Scarpa's ganglion by examining 35 human temporal bones by polymerase chain reaction. HSV-1 was found in 66% of geniculate ganglia and 60% of vestibular ganglia; all examined parts of vestibular ganglia were almost equally HSV-1 infected. Our data provided no support for viral migration along this anastomosis or for a preferential latency of HSV-1 in the superior vestibular nerve. We suggest that the common double innervation of the posterior ampulla by two nerves running in two separate bony canals could offer an alternative explanation for the regular sparing of posterior canal function in vestibular neuritis. PMID- 10482276 TI - Clinicopathological findings following intraventricular glial-derived neurotrophic factor treatment in a patient with Parkinson's disease. AB - As part of a safety and tolerability study, a 65-year-old man with Parkinson's disease (PD) received monthly intracerebroventricular injections of glial-derived neurotrophic factor (GDNF). His parkinsonism continued to worsen following intracerebroventricular GDNF treatment. Side effects included nausea, loss of appetite, tingling, L'hermitte's sign, intermittent hallucinations, depression, and inappropriate sexual conduct. There was no evidence of significant regeneration of nigrostriatal neurons or intraparenchymal diffusion of the intracerebroventricular GDNF to relevant brain regions. Alternative GDNF delivery systems should be explored. PMID- 10482277 TI - Analysis of gene expression in mutiple sclerosis lesions using cDNA microarrays. AB - In multiple sclerosis (MS) patients, a coordinated attack of the immune system against the primary constituents of oligodendrocytes and/or the myelin sheath of oligodendrocytes results in the formation of lesions in the brain and spinal cord. Thus far, however, a limited number of genes that potentially contribute to lesion pathology have been identified. Using cDNA microarray technology, we have performed experiments on MS tissue monitoring the expression pattern of over 5,000 genes and compared the gene expression profile of normal white matter with that found in acute lesions from the brain of a single MS patient. Sixty-two differentially expressed genes were identified, including the Duffy chemokine receptor, interferon regulatory factor-2, and tumor necrosis factor alpha receptor-2 among others. Thus, cDNA microarray technology represents a powerful new tool for the identification of genes not previously associated with the MS disease process. PMID- 10482279 TI - Acetylator genotype for N-acetyltransferase 2 and Parkinson's disease. PMID- 10482278 TI - The neurological manifestations of Nipah virus encephalitis, a novel paramyxovirus. AB - A novel Hendra-like paramyxovirus named Nipah virus (NiV) was the cause of an outbreak among workers from one abattoir who had contact with pigs. Two patients had only respiratory symptoms, while 9 patients had encephalitis, 7 of whom are described in this report. Neurological involvement was diverse and multifocal, including aseptic meningitis, diffuse encephalitis, and focal brainstem involvement. Cerebellar signs were relatively common. Magnetic resonance imaging scans of the brain showed scattered lesions. IgM antibodies against Hendra virus (HeV) were present in the serum of all patients. Two patients recovered completely. Five had residual deficits 8 weeks later. PMID- 10482280 TI - Acute bulbar weakness: thyrotoxicosis or myasthenia gravis? PMID- 10482281 TI - Reduction of transendothelial migration of mononuclear cells in interferon-beta1b treated multiple sclerosis patients. PMID- 10482282 TI - A stable prostacyclin-like substance produced by the medicinal leech Hirudo medicinalis. AB - The medicinal leech Hirudo medicinalis produces a low-molecular mass compound with properties similar to those of prostacyclin. It extracted with organic solvent, had affinity to 6-keto-PGF1alpha antibodies, inhibited human platelet aggregation induced in vitro by thrombin (by 50% at 4 pg/ml), and caused hypotension and secretion of plasminogen (t-PA) into the blood stream of rats. A main distinction from prostacyclin is stability of the substance due to covalent binding with the polypeptide chain of destabilase. Because of the high aggregability of destabilase, the molecules of the protein-lipid complex are organized into micelles that can change their spatial orientation depending on the nature of the solvent. Incorporation of hirudin and blood plasma kallikrein inhibitor into the micelle structure causes the formation of liposomes (with a molecular mass of the structural monomer 25 kDa). This complex with polypeptides provides not only stability but also rapid transmembrane penetration. The pure prostacyclin-like substance has a molecular mass of 391 Da and can be produced on destruction of the destabilase polypeptide chain. PMID- 10482283 TI - Expression of COX-1, COX-2, and inducible nitric oxide synthase protein in human gastric antrum with Helicobacter pylori infection. AB - For a better understanding of the regulation of prostaglandin and nitric oxide (NO) synthesis in circumstances in which the gastric mucosa is inflamed, we have examined the ex vivo production of NO and prostaglandin E2 and the protein expression of inducible nitric oxide synthase (iNOS) and 2 cyclo-oxygenase (COX) isoforms in gastric biopsies from nine Helicobacter pylori-infected patients with active gastritis and six Helicobacter pylori (HP)-negative patients. The results indicate a significant increased of NO and PGE2 in patients with HP infection compared with uninfected samples. These findings were paralleled by marked increases in iNOS and in COX-1 and COX-2 protein expression. Expression of iNOS and COX-2 protein was absent in the mucosa of HP-negative controls. We have demonstrated that iNOS protein is expressed in the gastric mucosa of patients with HP infection. It is likely that iNOS expression and the corresponding high release of NO may play an important role in gastric inflammation associated with HP infection. However, the expression of COX-1 and COX-2 and the parallel increase of prostaglandin E2 could imply that these factors could limit the extend of mucosal damage. In previous reports NO has been shown to stimulate the COX activity, so we think that the role of NO could be both in the regulation of normal function and in the genesis of diseases. PMID- 10482284 TI - Modulation of protein kinase C isoforms by PAF in cerebral cortex. AB - The effect of platelet activating factor (PAF) on subcellular distribution of protein kinase C isoforms in rat cerebral cortex was investigated. PAF induced an increase in levels of protein kinase C epsilon and gamma in membrane fraction. Results also indicate that PAF induced an increase in protein kinase C delta levels in both cytosolic and membrane fraction. This effect is possibly due to an increase in enzyme synthesis, as indicated by the results obtained from the experiments performed in the presence of cycloheximide and actinomycin. All the effects induced by PAF were time- and dose-dependent, and were mediated through the activation of PAF receptor. These findings indicate that the three isoforms may be involved in signal transduction of PAF in the brain. PMID- 10482285 TI - Effects of PAF on histamine H1 receptor mRNA expression in rat trigeminal ganglia. AB - The application of platelet-activating factor (PAF) to the nasal mucosa of humans has been shown to increase histamine-induced hyper-reactivity. To test the hypothesis that PAF acts by increasing the reactivity of sensory nerve endings in the nasal mucosa to histamine, we examined PAF-stimulated rat trigeminal nerve ganglion cells. We found that relatively low concentrations of PAF (10(-12)-10( 9) M) induced increased histamine H1 receptor mRNA expression. This increase appeared as early as 1 h after PAF stimulation, peaked at 4 h, and disappeared after 24 h. The PAF receptor antagonist WEB2086 inhibited the increased expression of histamine H1 receptor mRNA induced by PAF, suggesting that the effects of PAF are mediated by specific receptors. This PAF effect was abolished by actinomycin D, suggesting that PAF induces de novo transcription of histamine H1 and/or PAF receptor mRNA. PAF may be important in the hyper-responsiveness of nasal mucosa exposed to histamine. PMID- 10482286 TI - An imbalance between prostacyclin and thromboxane in relation to cerebral blood flow in neonates with maternal preeclampsia. AB - OBJECTIVE: A disturbance of prostacyclin (PGI2) and thromboxane A2 (TXA2) balance has been reported in preeclampsia. However, little is known about the concentrations of these prostanoids in neonates born to preeclamptic pregnant women. The purpose of this study is to determine whether the PGI2 and TXA2 concentrations are altered and whether the prostanoid balance correlates to the cerebral blood flow in neonates born to preeclampsia. METHODS: Spontaneously voided urine samples were collected from 20 neonates of normotensive and 16 neonates of preeclamptic women during the first 24 h after birth. We measured by radioimmunoassay the concentrations of urinary 6-keto-prostaglandin F1alpha (6 keto-PGF1alpha) and 11-dehydro-thromboxane B2 (11-dehydro-TXB2), respectively. Blood flow velocity in the middle cerebral artery was studied by pulsed Doppler ultrasonography in the neonates between 17 and 38 h after birth. RESULTS: There was no significant difference between the urinary 6-keto-PGF1alpha in the neonates of mothers with and without preeclampsia (median, 5.3 vs. 3.6 ng/mg of creatinine). In contrast, the urinary 11-dehydro-TXB2 and the ratio of 11-dehydro TXB2 to 6-keto-PGF1alpha in the neonates of mothers with preeclampsia were significantly lower as compared with the neonates without preeclampsia, respectively (13.7 vs. 20.6 ng/mg of creatinine and 3.0 vs. 5.2, median). The resistance index in the middle cerebral artery was significantly reduced in the neonates with preeclampsia than without preeclampsia (0.67 +/- 0.01 vs. 0.74 +/- 0.02, mean +/- SEM). CONCLUSIONS: There was an association between maternal preeclampsia and the imbalance in the neonatal urinary excretion of PGI2 and TXA2 metabolites. This imbalance may contribute to the regulation of cerebral blood flow. PMID- 10482287 TI - Decrease in thromboxane A2 receptor expression by differentiation with dibutyryl cyclic AMP in 1321N1 human astrocytoma cells. AB - Thromboxane A2 (TXA2) receptor expression with its signaling was investigated in 1321N1 human astrocytoma cells differentiated with dibutyryl cyclic AMP (dbcAMP). The cells cultured in 0.5% fetal calf serum containing 0.5 mM dbcAMP for 3 days showed the star-shaped morphology, accompanied with the reduction of a TXA2 mimetic U46619-induced phosphoinositide hydrolysis and Ca2+ mobilization. Immunoblotting analysis revealed that human astrocytoma cells expressed phospholipase C (PLC)-beta1 and -beta3, but not PLC-beta2. The contents of PLC beta1 and beta3 were not changed by the differentiation. The alpha subunit of Gq/ll bound to TXA2-receptor was reduced by the differentiation, determined by immunoblotting after immunoprecipitation with an anti-TXA2-receptor antibody. Scatchard analysis of the binding of [3H]SQ29548, a TXA2 receptor antagonist, to the membranes revealed that the maximum binding site was reduced by the differentiation. The expression of TXA2 receptor mRNA also was reduced by the differentiation, determined by reverse-transcribed-polymerase chain reaction. Although placental type of TXA2 receptor mRNA expression increased after the differentiation, endothelial type of TXA2 receptor mRNA expression slightly decreased. The results suggest that 1321N1 human astrocytoma cells differentiated with dbcAMP show impaired TXA2 receptor-mediated phosphoinositide hydrolysis and Ca2+ mobilization, due to the decrease in TXA2 receptor number. PMID- 10482288 TI - Solar disinfection: use of sunlight to decontaminate drinking water in developing countries. PMID- 10482289 TI - Erysipelothrix rhusiopathiae: bacteriology, epidemiology and clinical manifestations of an occupational pathogen. AB - Erysipelothrix rhusiopathiae has been recognised as a cause of infection in animals and man since the late 1880s. It is the aetiological agent of swine erysipelas, and also causes economically important diseases in turkeys, chickens, ducks and emus, and other farmed animals such as sheep. The organism has the ability to persist for long periods in the environment and survive in marine locations. Infection in man is occupationally related, occurring principally as a result of contact with animals, their products or wastes. Human infection can take one of three forms: a mild cutaneous infection known as erysipeloid, a diffuse cutaneous form and a serious although rare systemic complication with septicaemia and endocarditis. While it has been suggested that the incidence of human infection could be declining because of technological advances in animal industries, infection still occurs in specific environments. Furthermore, infection by the organism may be under-diagnosed because of the resemblance it bears to other infections and the problems that may be encountered in isolation and identification. Diagnosis of erysipeloid can be difficult if not recognised clinically, as culture is lengthy and the organism resides deep in the skin. There have been recent advances in molecular approaches to diagnosis and in understanding of Erysipelothrix taxonomy and pathogenesis. Two PCR assays have been described for the diagnosis of swine erysipelas, one of which has been applied successfully to human samples. Treatment by oral and intramuscular penicillin is effective. However, containment and control procedures are far more effective ways to reduce infection in both man and animals. PMID- 10482290 TI - Invasiveness of Salmonella serotypes Typhimurium, Choleraesuis and Dublin for rabbit terminal ileum in vitro. AB - Ten recent clinical isolates of Salmonella serotype Typhimurium from man that were tested for their invasiveness in rabbit ileal explants in vitro, were compared with Typhimurium strain TML, a well-characterised invasive strain isolated from a case of human gastro-enteritis. Nine of the 10 strains showed invasiveness that was comparable to that of strain TML. One isolate (GM3) was apparently substantially less invasive; electron microscopy showed this strain to be histotoxic - the probable reason for its reduced recovery from ileal mucosa and thus apparent 'low' invasiveness. Salmonella serotype Choleraesuis strain A50, isolated from a case of systemic salmonellosis in pigs, and serotype Dublin strain 3246, isolated from a case of systemic salmonellosis in calves, were also examined. Dublin strain 3246, when grown at 37 degrees C and used immediately in the invasion assay, damaged the mucosa in a manner similar to that of Typhimurium strain GM3, whereas Dublin strain 3246 grown at 37 degrees C and stored overnight at 4 degrees C did not. This was reflected in an apparently lower invasiveness of freshly grown organisms compared with that of organisms stored at 4 degrees C. In contrast, the histotoxicity of Typhimurium strain GM3 was not affected by storage at 4 degrees C. When stored at 4 degrees C, the levels of invasiveness of Choleraesuis strain A50 and Dublin strain 3246 were not significantly different from each other or from Typhimurium strain TML. PMID- 10482291 TI - A histotoxin produced by Salmonella. AB - Salmonella Typhimurium strain GM3, known to be histotoxic for explants of terminal rabbit ileum in vitro, produces similar lesions in vitro when sterile filtrates, obtained from live organisms after interaction with gut explants in vitro, are used and when rabbit ligated ileal loops are challenged with live organisms. Epithelial damage occurs rapidly, within 2 h of adding organisms or sterile filtrates. This evidence is construed in terms of a secreted salmonella histotoxin that causes epithelial damage, detaching enterocytes which rapidly degenerate into spheroid cells devoid of microvilli. Typhimurium strain GM3 invades ileal mucosa and bacteria are found in the subepithelial tissues. After 12 h, bacteria were seen to be expelled from infected villi in a manner similar to that seen in non-histotoxic infection with Typhimurium strain TML. PMID- 10482293 TI - Lipopolysaccharide chemotypes of Burkholderia cepacia. AB - Burkholderia cepacia is an important pathogen in patients with cystic fibrosis (CF) and much is now known of its epidemiology. In contrast, its virulence mechanisms are poorly understood. The lipopolysaccharide (LPS) of B. cepacia, a well-recognised virulence factor of other gram-negative bacteria, is known to be strongly endotoxic in vitro. The aim of this study was to observe if there were any links between the structure of B. cepacia LPS and virulence. This has been investigated by polyacrylamide gel electrophoresis and immunoblotting to define the chemotype and antigenic cross reactivity of B. cepacia LPS. Strains (16) belonging to different genomovars of the B. cepacia complex were selected to represent epidemic and non-epidemic clinical isolates and environmental strains. All strains belonging to genomovars I and II (the latter now renamed B. multivorans) had smooth LPS. However, isolates belonging to genomovar III, the group to which most of the epidemic CF isolates belong - including the highly transmissible strain (ET 12) which has been found in both the UK and North America - were of either rough or smooth LPS chemotype. In this study, B. cepacia J2315 represents the ET 12 lineage, and has a rough chemotype. Rabbit antiserum raised to strain J2315 revealed that the LPS core of this strain was antigenically related to some but not all other genomovar III strains, but it also cross-reacted strongly with all B. multivorans (genomovar II) and most genomovar I strains. Intra-strain phenotypic variation was demonstrated between bacteria grown in broth or on solid agar with a concomitant variation in antigenic cross reactivity. There was no clear evidence to associate any particular LPS phenotype with epidemic or non-epidemic strains, but changes in phenotype in vitro may provide clues to the survival and adaptability of B. cepacia in hostile environments and possibly to its ability to produce an inflammatory response in vivo. PMID- 10482292 TI - Enterotoxin production by coagulase-negative staphylococci in restaurant workers from Kuwait City may be a potential cause of food poisoning. AB - Staphylococcus aureus and coagulase-negative staphylococci (CNS) were isolated from the hands of food handlers in 50 restaurants in Kuwait City and studied for the production of staphylococcal enterotoxins, toxic shock syndrome toxin-1, slime and resistance to antimicrobial agents. One or a combination of staphylococcal enterotoxins A, B or C were produced by 6% of the isolates, with the majority producing enterotoxin B. Toxic shock syndrome toxin-1 was detected in c. 7% of the isolates; 47% produced slime. In all, 21% of the isolates were resistant to tetracycline and 11.2% were resistant to propamidine isethionate and mercuric chloride. There was no correlation between slime and toxin production or between slime production and antibiotic resistance. The detection of enterotoxigenic CNS on food handlers suggests that such strains may contribute to food poisoning if food is contaminated by them and held in conditions that allow their growth and elaboration of the enterotoxins. It is recommended that enterotoxigenic CNS should not be ignored when investigating suspected cases of staphylococcal food poisoning. PMID- 10482295 TI - Phylogenetic analysis of Calymmatobacterium granulomatis based on 16S rRNA gene sequences. AB - Calymmatobacterium granulomatis is the aetiological agent of granuloma inguinale a chronic granulomatous genital infection - and is morphologically similar to members of the genus Klebsiella. This study determined the 16S rRNA gene sequence of C. granulomatis and the taxonomic position of the organism in relation to the genus Klebsiella. Genomic DNA was extracted from C. granulomatis-infected monocytes and from frozen and formalin-fixed paraffin wax-embedded tissue biopsy specimens from patients with histologically proven granuloma inguinale. The 16S rDNA was amplified by PCR with broad range oligonucleotide primers. The amplified DNA fragments were cloned into pMOS vector, digested with Bam HI and Pst1 restriction endonucleases, hybridised with a gram-negative bacterial probe (DL04), sequenced in both directions by the automated ALF DNA sequencer, verified on an ABI Prism 377 automated sequencer and analysed with DNASIS and MEGA software packages. Sequence analysis revealed DNA homology of 99% in C. granulomatis from the different sources, supporting the belief that the bacteria in the culture and the biopsy specimens belonged to the same species, although there was some diversity within the species. Phylogenetically, the strains were closely related to the genera Klebsiella and Enterobacter with similarities of 95% and 94% respectively. C. granulomatis is a unique species, distinct from other related organisms belonging to the gamma subclass of Proteobacteria. PMID- 10482294 TI - The 18-kDa cytoplasmic protein of Brucella species --an antigen useful for diagnosis--is a lumazine synthase. AB - Previous studies have shown that the detection of antibodies to an 18-kDa cytoplasmic protein of Brucella spp. is useful for the diagnosis of human and animal brucellosis. This protein has now been expressed in recombinant form in Escherichia coli. The recombinant protein is soluble only under reducing conditions, but alkylation with iodoacetamide renders it soluble in non-reducing media. As shown by gel exclusion chromatography, this soluble form arranges in pentamers of 90 kDa. The reactivity of human and animal sera against the recombinant protein was similar to that found with the native protein present in brucella cytoplasmic fraction, suggesting that the recombinant protein is correctly folded. The protein has low but significant homology (30%) with lumazine synthases involved in bacterial riboflavin biosynthesis, which also arrange as pentamers. Biological tests on the crude extract of the recombinant bacteria and on the purified recombinant protein showed that the biological activity of the Brucella spp. 18-kDa protein is that of lumazine synthase. Preliminary crystallographic analysis showed that the Brucella spp. lumazine synthase arranges in icosahedric capsids similar to those formed by the lumazine synthases of other bacteria. The high immunogenicity of this protein, potentially useful for the design of acellular vaccines, could be explained by this polymeric arrangement. PMID- 10482296 TI - Seroprevalence of Bartonella henselae in cats in Germany. AB - Bartonella henselae and B. quintana infections in man are associated with various clinical manifestations including cat-scratch disease, bacillary angiomatosis and bacteraemia. While cats are the natural reservoir for B. henselae, the source of B. quintana is unclear. In this study, the sera of 713 cats from Germany were examined for the presence of antibodies against B. henselae, B. quintana or Afipia felis by an indirect immunofluorescence assay (IFA). Bartonella-specific antibody titres of > or =50 were found in 15.0% of the cats. There was substantial cross-reactivity among the various Bartonella antigens, although single sera showed high titres against B. henselae but not against B. quintana and vice versa. Antibodies against A. felis were not detected in any of these cats. Statistical analysis indicated that there is no correlation between Bartonella infections and the sex, age or breed of the cat or its hunting behavior. There was also no correlation between bartonella and toxoplasma infections in cats. However, whereas 16.8% of cats from northern Germany had B. quintana-specific antibodies, only 8.0% of cats from southern Germany were seropositive for B. quintana. No statistically significant difference was found for B. henselae. IFA-positive and IFA-negative sera were used for immunoblot analysis including B. henselae and B. quintana. Marked reactivity was observed with protein bands at 80, 76, 73, 65, 37, 33 and 15 kDa. The results of this study suggest that B. henselae, and possibly a B. quintana-related pathogen, but not A. felis, are common in cats in Germany, and that there are differences in the geographic distribution of bartonella infections in cats. PMID- 10482297 TI - Design of a one-tube hemi-nested PCR for detection of Toxoplasma gondii and comparison of three DNA purification methods. AB - The aims of the present study were to design an easy and sensitive DNA amplification method for detection of Toxoplasma gondii with low risk of accidental contamination, and to find a rapid method for purification of clinical samples containing potential inhibitors of the amplification reaction. With a pair of primers amplifying a 619-bp fragment of the B1 gene of this parasite it was possible to detect DNA equivalent to 10 parasites. When a third primer was added to the same tube, sensitivity increased to 0.1 parasite. In a comparison of different DNA purification methods, the High Pure PCR Template Preparation Kit (Boehringer Mannheim, Germany) gave the best results. With this purification method and the one-tube hemi-nested PCR, T. gondii DNA was detected in 14 (87.5%) of 16 clinical specimens (amniotic fluid, broncho-alveolar lavage, bone marrow, blood, liver biopsy) in which the parasite was demonstrated by cell culture. PMID- 10482298 TI - Detection of pneumolysin in sputum. AB - Western blot detection of the species-specific pneumococcal product, pneumolysin (SPN), was shown to be almost as sensitive as PCR for the non-cultural detection of pneumococci in 27 Streptococcus pneumoniae culture-positive sputa from patients stated to have chest infections. Both techniques were considerably more sensitive than counter-current immuno-electrophoresis for pneumococcal capsular polysaccharide antigens (CPS-CIE) on the same specimens. Sensitivities for PCR, SPN-immunoblotting and CPS-CIE were 100%, 85% and 67%, respectively. In 11 S. pneumoniae culture-negative sputa taken from patients receiving antibiotics, but with proven recent pneumococcal infection, PCR and SPN-blot were positive in six (in two of which CPS-CIE was also positive), PCR alone was positive in one and SPN-blot alone was positive in one. In 11 S. pneumoniae culture-negative samples from patients not receiving antibiotics, all three tests were negative in eight, PCR was positive in three (in one of which CPS-CIE was also positive), but SPN blot was negative in all 11. In 16 S. pneumoniae culture-negative samples from patients receiving antibiotics and with no known recent pneumococcal infections, one or more non-cultural test was positive in 11. Although further evaluation is required to assess the significance of pneumolysin detection in relation to carriage and infection and to devise a more suitable test format, these preliminary studies suggest that pneumolysin detection is a promising new approach to the non-cultural diagnosis of pneumococcal chest infection. PMID- 10482299 TI - Elucidation of the antistaphylococcal action of lactoferrin and lysozyme. AB - The cationic tear proteins lactoferrin and lysozyme exhibit co-operative antistaphylococcal properties. The purpose of this study was to determine the mechanism of action of this co-operation on Staphylococcus epidermidis. Following blocking of lipoteichoic acid (LTA) binding sites, the effects on binding of lactoferrin and susceptibility to lactoferrin and lysozyme were determined. The effect of lactoferrin on autolysis and LTA release was also examined. Maximal susceptibility occurred on addition of lactoferrin first followed by lysozyme. Blocking the LTA binding sites both reduced lactoferrin binding and decreased susceptibility. Autolytic activity decreased and LTA release increased in the presence of lactoferrin. These results suggest that binding of lactoferrin to LTA is important in its synergy with lysozyme and interferes with the autolysins present on the LTA. It is proposed that, on binding to the anionic LTA of S. epidermidis, the cationic protein lactoferrin decreases the negative charge, allowing greater accessibility of lysozyme to the underlying peptidoglycan. PMID- 10482300 TI - Cross-reaction between a strain of Vibrio mimicus and V. cholerae O139 Bengal. AB - Of 200 isolates of Vibrio mimicus screened, one from water (N-57) agglutinated with V. cholerae O139 polyclonal antiserum (absorbed with a rough strain of V. cholerae only) and not with O139 polyclonal diagnostic antiserum (absorbed with the rough strain and V. cholerae O22 and O155). The antigenic relationship between V. cholerae 0139 and N-57 is of a,b-a,c type, where a is the common antigenic epitope and b and c are unique epitopes. Strain N-57 was assigned to a new serogroup of V. cholerae O194. It gave negative results in a monoclonal antibody-based rapid test and a PCR test specific for V. cholerae O139. It did not possess the ctx gene or produce cholera toxin. Antiserum to strain N-57 cross protected infant mice against cholera on challenge with V. cholerae O139. Structural studies of the surface polysaccharides and studies of the rfb genes will shed more light on the extent of relatedness between V. mimicus N-57 and V. cholerae O139. PMID- 10482301 TI - Medical advice editorials: transferring medical science into clinical practice. PMID- 10482302 TI - Could apoptosis be responsible for localized imbalances in bone cell homeostasis? PMID- 10482303 TI - Do diuretics cause thiamine deficiency? PMID- 10482304 TI - Somatic gene therapy for dyslipidemias. AB - Somatic gene transfer is a valuable tool for the in vivo evaluation of lipoprotein metabolism. It has been used to dissect metabolic pathways, to establish structure-function relationships of various gene products, and to evaluate conventional lipid-lowering and novel therapeutic genes for the treatment of lipoprotein disorders. In this article we review some general aspects of somatic gene therapy and the different vehicles used for the delivery of therapeutic genes. We highlight some recent advances in adenoviral vector development that make this vector an attractive system for clinical trials. PMID- 10482305 TI - Quality control in the endoplasmic reticulum: lessons from hereditary myeloperoxidase deficiency. AB - The optimal level of oxygen-dependent microbicidal activity in human neutrophils depends on the generation of highly toxic products, including hypochlorous acid, by hydrogen peroxide in the presence of chloride anion and the neutrophil granule protein myeloperoxidase (MPO). The biosynthesis of MPO is normally restricted to the promyelocytic stage of myeloid development and includes N-linked glycosylation, heme insertion, proteolytic processing, subunit dimerization, and eventual targeting to the azurophilic granule. In the endoplasmic reticulum, MPO precursors interact transiently with calreticulin and calnexin, presumably in their capacity as molecular chaperones. In light of the important role of the MPO H2O2-chloride system in human host defense, the relatively high prevalence of inherited MPO deficiency was an unanticipated insight provided by the widespread use of automated flow cytometry for the enumeration of leukocytes in clinical specimens. In many cases of inherited MPO deficiency, affected neutrophils have immunochemical evidence of precursor protein but lack the subunits of mature MPO, peroxidase activity, or the ability to chlorinate target proteins. To date, four genotypes have been reported to cause inherited MPO deficiency, each of which results in missense mutations. In the genotype Y173C, the mutant precursor is retained in the endoplasmic reticulum by virtue of its prolonged interaction with calnexin, and it eventually undergoes degradation in the 20S proteasome. In this way, the quality control system operating in the endoplasmic reticulum retrieves malfolded MPO precursors from the biosynthetic pathway and creates the biochemical phenotype of MPO deficiency. Thus MPO deficiency caused by Y173C joins the ranks of cystic fibrosis, protein C deficiency, and other genetic disorders that reflect abnormalities in protein folding. PMID- 10482306 TI - Tumor necrosis factor-alpha and interleukin-1beta increase the Fas-mediated apoptosis of human osteoblasts. AB - Our recent work demonstrated functional Fas expression on human osteoblasts, and the histologic examination of the periarticular osteoporosis region in patients with rheumatoid arthritis (RA) showed apoptosis in osteoblasts. High concentrations of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF alpha), and IL-6--which are thought to increase bone resorption--have been determined in RA synovium. We investigated the effect of these cytokines on the Fas-mediated apoptosis of human osteoblasts. The human osteoblastic cell line MG63 and human primary osteoblast-like cells from bone biopsy specimens were used as human osteoblasts. Fas expression on these cells was examined by flow cytometry, and Fas-mediated apoptosis induced by anti-Fas immunoglobulin M (IgM) was determined by a chromium 51 release assay, the presence of cells with hypodiploid DNA, staining with Hoechst 33258 dye, and the detection of DNA fragmentation on agarose gel electrophoresis. The proliferation of osteoblasts was analyzed by a tritiated thymidine incorporation assay. Spontaneous apoptosis was not found on cultured osteoblasts. The apoptosis of human osteoblasts was not induced by TNF-alpha, IL-1beta, or IL-6 alone in the absence of anti-Fas IgM. In addition, proliferation of the cells was not affected by these cytokines. Fas was constitutively expressed on unstimulated osteoblasts, and treatment of these cells with IL-1beta or TNF-alpha significantly augmented Fas expression. Human osteoblasts were committed to apoptosis with anti-Fas IgM, and the treatment of both IL-1beta and TNF-alpha markedly increased Fas-mediated apoptosis. TNF-alpha augmented both Fas expression and Fas-mediated apoptosis more efficiently than did IL-1beta. In addition, an additive effect on both Fas expression and Fas mediated apoptosis was demonstrated when TNF-alpha and IL-1beta were added to osteoblasts. IL-6 influenced neither Fas expression nor the Fas-mediated apoptosis of osteoblasts. Furthermore, no synergistic effect of IL-6 with IL 1beta or TNF-alpha was observed. IL-1beta, TNF-alpha, or IL-6 did not change Bcl 2 expression. Our results suggest that IL-1beta and TNF-alpha regulate osteoblast cell number by up-regulating the Fas-mediated apoptosis of osteoblasts, one of the putative mechanisms inducing periarticular osteoporosis in patients with RA. PMID- 10482307 TI - Urinary thiamine excretion in the rat: effects of furosemide, other diuretics, and volume load. AB - Long-term furosemide therapy is associated with increased urinary loss of thiamine. To examine the mechanism of furosemide-induced urinary thiamine loss, we measured urinary excretion of thiamine in rats in response to increasing doses of furosemide, acetazolamide, chlorothiazide, amiloride, mannitol, and extracellular fluid (ECF) volume loading by saline infusion. All animals were in normal thiamine balance as reflected by a thiamine pyrophosphate effect (TPPE) of 2.25% +/- 0.60% (mean +/- SEM), and all had normal renal function. Urinary flow increased in response to diuretic administration in a dose-dependent manner, reaching (mean) peak urinary flow rates of 283 to 402 microL/min. Fractional excretion of sodium (FE(Na)) exhibited the same pattern, reaching peak values of 12.3% to 23.2%. Urinary thiamine excretion increased in proportion to the incremental doses of diuretic agents, reaching (mean) maximal values of 7.44 to 9.34 pmol/min, with no significant difference (P = .11) between the various diuretics tested nor in response to saline loading. None of the diuretics tested differed in the effect on thiamine excretion, which was clearly flow dependent and only partially related to fractional sodium excretion. Urinary flow rate, being the single significant predictor, explained 78% (R2 = 0.78) of the variability in thiamine excretion rates. These findings indicate that urinary thiamine loss is caused by a nonspecific, flow-dependent mechanism common to all of the diuretics tested. PMID- 10482308 TI - Urinary loss of thiamine is increased by low doses of furosemide in healthy volunteers. AB - Prolonged furosemide treatment is associated with urinary loss of thiamine and thiamine deficiency in some patients with congestive heart failure and low dietary thiamine intake. In the rat, diuretic-induced thiamine urinary loss is solely dependent on increased diuresis and is unrelated to the type of diuretic used. We studied the effects of single intravenous doses of furosemide (1, 3, and 10 mg) and of normal saline infusion (750 mL) on urinary thiamine excretion in 6 volunteers. Over a 6-hour period, furosemide induced dose-dependent increases in urine flow and sodium excretion rates (mean +/- SD), from 51 +/- 17 mL/h at baseline to 89 +/- 29 mL/h, 110 +/- 38 mL/h, and 183 +/- 58 mL/h (F = 10.4, P < .002) and from 5.1 +/- 2.3 mmol/h to 9.4 +/- 6.8 mmol/h, 12.1 +/- 2.6 mmol/h, and 20.9 +/- 10.6 mmol/h (F = 6.3, P < .005) for the three doses, respectively (104 +/- 35 mL/h and 13.0 +/- 6.2 mmol/h for the saline infusion). During this period the thiamine excretion rate doubled from baseline levels (mean of four 24-hour periods before the diuretic interventions) of 6.4 +/- 5.1 nmol/h to 11.6 +/- 8.2 nmol/h (F = 5.03, P < .01, for all four interventions, no difference being found between them), then returning over the following 18 hours to 6.1 +/- 3.9 nmol/h. The thiamine excretion rate was correlated with the urine flow rate (r = 0.54, P < .001), with no further effect of the type of intervention or sodium excretion rate. These findings complement our previous results in animals and indicate that sustained diuresis of >100 mL/h induces a nonspecific but significant increase in urinary loss of thiamine in human subjects. Thiamine supplements should be considered in patients undergoing sustained diuresis, when dietary deficiency may be present. PMID- 10482310 TI - Evidence for interference in estradiol-17beta inactivation to estrone by oxidized low-density lipoprotein and selected lipid peroxidation products. AB - An elevation in plasma estrogen levels is believed to play a key role in the pathogenesis of breast cancer. The conversion of estradiol-17beta (E2) to estrone (E1) by 17beta-hydroxy steroid dehydrogenase type 4 (17-HSD4) represents a major pathway of its inactivation in cells. In this study the potential relationship between lipoprotein peroxidation products and E2 metabolism was examined. It was noted that oxidized low-density lipoprotein (OX-LDL), not native LDL, caused a time- and concentration-dependent inhibition of the conversion of labeled E2 to E1 in THP-1 macrophage cells. Further studies noted that among the lipoprotein peroxidation products examined, malondialdehyde (MDA) caused a marked decrease in this reaction, whereas hexanal and a variety of oxysterols had no effect. This inhibition of E1 formation from E2 in THP-1 cells was confirmed by the quantitation of estrone formed with high-pressure liquid chromatography and by the expression of 17-HSD4 by reverse transcriptase-polymerase chain reaction. MDA added to Hep G2 cells showed a similar trend in E1 formation. These results suggest that increased oxidative stress and lipid peroxidation might result in decreased inactivation of biologically active estrogen. This might be important in postmenopausal women undergoing estrogen replacement therapy. PMID- 10482309 TI - A double-blind, placebo-controlled study of Mycobacterium-specific human immune responses induced by intradermal bacille Calmette-Guerin vaccination. AB - Recent studies have indicated that type 1 T cell responses (potent interferon gamma and cytolytic responses, with absence of interleukin-4 production) are important for protective immunity against mycobacteria. These observations suggest that assays of type 1 T cell responses may be useful as surrogate markers of protective immunity in the evaluation of new tuberculosis vaccines. To be useful as surrogate markers, immunologic assays must distinguish between vaccine recipients and control subjects in clinical trials. Previous studies have shown that bacille Calmette-Guerin (BCG) vaccination can induce human type 1 T cell responses, but randomized trials have not been done to determine whether measurement of these responses can distinguish between BCG recipients and control subjects. We have conducted a double-blind, placebo-controlled trial of intradermal vaccination with two different BCG strains. We compared the mean lymphoproliferative, cytotoxic, Th1 and Th2 cytokine, and antibody responses detected in BCG and placebo recipients. These studies demonstrated that significant increases in Mycobacterium-specific T cell proliferative responses and type 1 cytokine responses were induced by BCG when compared with results with a placebo. In addition, BCG induced significant increases in Mycobacterium specific antibody responses with an isotype profile characteristic of a type 1 cytokine bias. T cell and antibody assays involving the use of mycobacterial whole cell lysates or live BCG were able to discriminate between BCG and placebo recipients better than were assays using mycobacterial culture filtrates. These studies provide important information for the development of immunologic assays that might be useful as surrogate markers of protective immunity in future trials of new tuberculosis vaccines. PMID- 10482311 TI - Plasminogen II accumulates five times faster than plasminogen I at the site of a balloon de-endothelializing injury in vivo to the rabbit aorta: comparison with other hemostatic proteins. AB - In the rabbit blood stream, plasminogen circulates as two glycoforms, plasminogen I (PLG-I) and plasminogen II (PLG-II), in a molar ratio of 1:2.2. To compare their relative behaviors toward a site of vascular injury, radiolabeled samples of PLG-I and PLG-II were coinjected intravenously into NZW rabbits before inducing a de-endothelializing (balloon catheter) injury to the thoracic aorta. At various times (5 to 60 minutes) after injury, each rabbit was anesthetized and exsanguinated, the aorta was excised, and the radioactivity per centimeters squared of aortic intima-media (IM) was measured relative to that of blood at exsanguination. The uptake of iodine 125-labeled PLG-I and iodine 131-labeled PLG II showed that the IM was essentially saturated by both glycoforms by 30 to 40 minutes after injury. Extrapolation of the flux rates to 1 minute after injury indicated that the uptake of PLG-II (2.4 pmol/min/cm2) exceeded PLG-I (0.5 pmol/min/cm2) almost five-fold. This result is consistent with an earlier report (Metabolism 1994;43:1430-7) that PLG-II is released by the liver and catabolized in vivo approximately five times faster than PLG-I. By molar comparison, the flux of total plasminogen (ie, PLG-I plus PLG-II) into the injured aorta wall in vivo was 2.4 times greater than that for prothrombin. Assuming both zymogens are converted to their respective proteases within the wound site, then approximately 2 to 3 molecules of plasmin are released for each molecule of thrombin in vivo. The possible significance of this plasmin:thrombin ratio is discussed in respect to the turnover of fibrin(ogen) within the site of vascular injury. PMID- 10482312 TI - Pregnancy aggravates proteinuria in subclinical glomerulonephritis in the rat. AB - Because subclinical renal disease may be aggravated during pregnancy--as reflected in the occurrence of proteinuria, for example--we investigated whether a subclinical glomerulonephritis (SG) in the non-pregnant rat (passive Heymann nephritis), a condition without proteinuria, is aggravated when the animals become pregnant and, if so, whether this is associated with a glomerular inflammatory reaction. SG was induced in non-pregnant rats 8 days before pregnancy. On day -1, part of the group of rats became pregnant. Three experiments were performed. In experiment 1, 4-hour urine albumin excretions and blood pressure (tail cuff) were measured. In experiment 2, the glomerular filtration rate (GFR) was measured with the chromium 51-labeled ethylenediaminetetraacetic acid method, while in experiment 3, parameters characteristic of a glomerular inflammation were studied. Experiment 1 revealed that non-pregnant rats with SG did not exhibit proteinuria. However, after the rats became pregnant, a significant proteinuria occurred, without an increase in systolic blood pressure. Experiment 2 revealed that the GFR did not increase in pregnant rats with SG, while experiment 3 showed that only pregnant animals exhibited a significant glomerular inflammation; this glomerular inflammation was characterized by intraglomerular influx of polymorphonuclear cells and monocytes. The results suggest that an SG in the rat may flare up during pregnancy. This exacerbation is characterized by proteinuria and coincides with a glomerular inflammatory reaction. It is suggested that proteinuria and the glomerular inflammatory reaction are causally related and promoted by the pregnant condition. PMID- 10482314 TI - Angiotensin II infusion exacerbates radiation nephropathy. AB - We hypothesized that angiotensin II will exacerbate radiation nephropathy in a time-specific manner. Experimental radiation nephropathy is treatable with angiotensin-converting enzyme inhibition or angiotensin II (AII) receptor blockers. These interventions are particularly important between 3 and 10 weeks after irradiation. We therefore undertook studies in which AII infusions were given at particular intervals after irradiation. Rats received total body irradiation (TBI) plus syngeneic bone marrow transplantation followed (or not) by AII infusion at 200 or 400 ng/kg/min. Infusions were given from 0 to 4 or 4 to 8 weeks after irradiation. An additional group was unirradiated but infused at 800 ng/kg/min for 8 weeks. Kidney function was assessed over 26 weeks, and histology was evaluated after the animals were killed. AII infusion alone did not cause azotemia. There was transient hypertension during AII infusion at 800 ng/kg/min but only minor histologic injury. Irradiation caused azotemia and hypertension, which were not exacerbated by AII infusion at 200 ng/kg/min. Irradiation plus AII infusion at 400 ng/kg/min from 4 to 8 weeks after TBI caused significantly greater azotemia than irradiation alone or irradiation with AII infusion from 0 to 4 weeks. The blood pressure was higher in irradiated rats infused with AII from 4 to 8 weeks. Arteriolar fibrinoid necrosis was a prominent feature in kidneys of rats infused with AII from 4 to 8 weeks after TBI. The worsening of radiation nephropathy by AII infusion from 4 to 8 weeks after irradiation strongly supports the idea of specific and sequential events in the pathogenesis of kidney failure in this model. Hypertension may play a role in these events in addition to the effect of AII alone. The occurrence of arteriolar fibrinoid necrosis in the irradiated, 4-to-8-week-infused animals suggests that vascular injury during that interval determines later outcome. PMID- 10482313 TI - Despite structural similarities between gp91phox and FRE1, flavocytochrome b558 does not mediate iron uptake by myeloid cells. AB - Superoxide (O2-) generated by the phagocyte reduced nicotinamide adenine dinucleotide phosphate oxidase is dependent on electron transfer by flavocytochrome b558 (flavocytochrome b), a transmembrane heterodimer that forms the redox center of the oxidase at the plasma or phagosomal membrane. The larger of its two subunits, gp91phox, is homologous to the yeast iron reductase subunit FRE1, and these two proteins share many structural and functional characteristics. Because FRE1 is required for iron uptake in yeast, we hypothesized that flavocytochrome b might serve a similar function in human phagocytes and thus provide a mechanism for the transferrin-independent iron acquisition observed in myeloid cells. To determine whether flavocytochrome b was required for iron uptake, we compared iron acquisition by polymorphonuclear neutrophils (PMNs) or Epstein-Barr virus (EBV)-transformed B lymphocytes derived from individuals with X-linked chronic granulomatous disease (CGD) with iron acquisition by normal cells. Our results indicate that all cells acquired iron to the same extent and that uptake could be significantly enhanced in the presence of the trivalent metal gallium. The gallium enhancement of iron uptake observed in PMNs or in EBV-transformed B lymphocytes derived from healthy individuals was mirrored by those derived from individuals deficient in flavocytochrome b. Furthermore, both normal and CGD-derived EBV-transformed B lymphocytes had similar iron reductase activity, suggesting that flavocytochrome b is not a biologically significant iron reductase. In contrast to previously suggested hypotheses, these results show conclusively that flavocytochrome b is not necessary for cellular iron acquisition, despite structural and functional similarities between yeast iron reductases and flavocytochrome b. PMID- 10482315 TI - Effect of renovascular hypertension on experimental glomerulonephritis in rats. AB - Systemic hypertension is a major risk factor that determines the rate of progression of kidney disease. The underlying mechanisms, however, are incompletely understood. To gain insight into these mechanisms, the present study was undertaken to characterize the effects of renovascular hypertension on the course of anti-thymocyte antibody-induced glomerulonephritis. Glomerulonephritis was induced in rats 6 weeks after the initiation of two-kidney, one-clip hypertension, when blood pressure was already increased. Structure and function of the clipped and the nonclipped kidney were examined 5 days later. Glomerular filtration rate (GFR) was measured by inulin clearance. The induction of nephritis did not alter the blood pressure in either hypertensive rats or normotensive controls. Albuminuria increased slightly in normotensive rats after the induction of nephritis, whereas no significant differences were found between hypertensive rats with or without nephritis. No significant differences were found for the GFR values of normotensive controls and nephritic animals or for values in the clipped kidney with or without nephritis. However, the GFR of the nonclipped kidney was significantly reduced in nephritic animals as compared with all other groups. Morphologic evaluation revealed that hypertensive rats with nephritis exhibited a combination of characteristics of nephritis and hypertensive glomerular injury. Histologic findings of nephritis, such as glomerular binding of rabbit IgG and glomerular proliferation and mesangial matrix expansion, were similar after the induction of nephritis in controls and in the clipped and nonclipped kidneys of hypertensive animals. However, intraglomerular microaneurysms were significantly more often found in the non clipped kidneys after the induction of nephritis. Hypertension-induced deterioration of glomerular function was not associated with marked morphologic deterioration but rather with a combination of the characteristics of nephritis and hypertensive glomerular injury. PMID- 10482316 TI - Augmentation of kidney injury by basic fibroblast growth factor or platelet derived growth factor does not induce progressive diabetic nephropathy in the Goto Kakizaki model of non-insulin-dependent diabetes. AB - Diabetes is now the most common cause of kidney failure. The pathogenesis of diabetic nephropathy in both type 1 and type 2 diabetes, however, is still incompletely understood. Two mechanisms postulated to contribute to the pathogenesis of progressive diabetic nephropathy are glomerular cell proliferation and glomerular visceral epithelial cell or podocyte injury. The aim of the current study was to determine whether the aggravation of mesangial cell injury or podocyte injury in isolation would induce progressive diabetic nephropathy. Specifically, we examined whether the administration of either platelet-derived growth factor (PDGF) or basic fibroblast growth factor (bFGF) in sub-nephritogenic doses might lead to an aggravation of kidney structural changes associated with hyperglycemia, resulting in progressive kidney damage in the Goto Kakizaki (GK) rat, which is a genetic model of non-obese non-insulin-dependent diabetes mellitus (NIDDM), in which progressive kidney disease does not develop spontaneously. The results demonstrate that the administration of PDGF to hyperglycemic GK rats led to acute mesangial cell proliferation and activation as assessed by 5-bromo-2'-deoxyuridine-positive nuclei and immunostaining for alpha smooth muscle actin. Despite acute mesangial cell activation and proliferation, PDGF treatment had no long-term effect on either kidney structure or function. Similarly, treatment of GK rats with bFGF, despite the augmentation of podocyte injury as demonstrated by de novo expression of glomerular desmin expression, did not lead to the development of progressive diabetic nephropathy. In summary, the data in the current manuscript suggest that the additive effect of hyperglycemia and superimposed isolated mesangial cell or podocyte injury does not lead to progressive diabetic nephropathy. This further emphasises the multifactorial nature of the pathogenesis of progressive diabetic nephropathy. PMID- 10482317 TI - DNA sequence of the canine platelet beta3 gene from cDNA: comparison of canine and mouse beta3 to segments that encode alloantigenic sites and functional domains of beta3 in human beings. AB - The platelet glycoprotein complex alphaIIb beta3 is required for platelet fibrinogen binding and platelet aggregation. This study was designed to characterize the nucleotide sequence of the canine platelet beta3 gene from cDNA. The nucleotide and deduced amino acid sequences of the canine beta3 gene were 92% and 96% homologous, respectively, with the sequences previously established for the beta3 gene of human beings. Within the beta3 gene, the nucleotide sequence of cDNA prepared from canine platelets shared homology of 89% for the cytoplasmic domain, 93% for the transmembrane domain, 92% for the extracellular domain, 94% for the arginine-glycine-aspartic acid (RGD) binding domain, and 97% for the region associated with Ca2+-dependent stabilization of the alphaIIb beta3 fibrinogen-binding pocket. The deduced amino acid sequence of canine beta3 was 100%, 97%, 96%, and 95% homologous with the cytoplasmic, transmembrane, extracellular, and RGD-binding domains, respectively, and was 100% homologous with the region associated with Ca2+-dependent stabilization of the alphaIIb beta3 fibrinogen-binding pocket of beta3 in human beings. The canine platelet cDNA signal peptide segment of the beta3 gene encodes for 22 amino acids, as compared with 26 amino acids previously reported for human beings. The deduced amino acid sequence of canine beta3 corresponds to the high-frequency allelic form for five of the six alloantigenic sites reportedly associated with human platelets: Leu33Leu40Pro407Arg489Arg636. The apparent amino acid residue in position 143 (Pen alloantigen) of canine platelet beta3 is histidine compared with arginine in human beings. Knowledge of the beta3 gene nucleotide sequence of normal dogs will facilitate the understanding of platelet alphaIIb beta3 structure-function relationships. PMID- 10482319 TI - Development of a gutless vector. PMID- 10482318 TI - Treatment of Wilson's disease with zinc. PMID- 10482320 TI - Re-engineering an Australian emergency department: can we measure success? AB - In 1996, in response to perceived deficiencies of the Emergency Department, Sir Charles Gairdner Hospital made emergency medicine a key strategic initiative. Major staffing and functional changes occurred as a result, including creation of the first Chair in Emergency Medicine in Australasia. We present a before and after study, using a range of measured variables, including the accepted Australian Council on Healthcare Standards emergency medicine clinical indicators. Clinically, there were great improvements in waiting times, time to thrombolysis in acute myocardial infarction, complaint rate, and misdiagnosed fracture rate. Increased throughput of short stay patients in a re-opened observation ward greatly shortened average length of stay for patients with a range of acute conditions. Data also indicated significant improvements in teaching and research. We conclude that with firm commitment from hospital management, re-engineering an emergency department can be shown to improve the quality-of-care. PMID- 10482321 TI - Customer focus: patient, organization and EQuIP in collaboration. Evaluation and Quality Improvement Program. AB - The Australian Council on Healthcare Standards (ACHS) Evaluation and Quality Improvement Program (EQuIP) calls on healthcare organizations to increase their focus on patients by using leadership to coordinate, and continuous improvement to guide, care delivery. At a large acute care private facility in Melbourne, a program has been developed to create a 'care partnership', characterized by shared decision making, collaboration and conciliation. This program enhances patient care through the coordination of three strategies, a patient communication strategy, an evaluation strategy and a quality improvement strategy. The program has resulted in patient guided reforms such as redesign of a patient information booklet, a hospital-wide discharge planning improvement initiative and a hospital-wide strategy to improve pain management. Through the creation of a care partnership, this program helps an acute care hospital focus its services and processes on one of its key customer groups, patients. PMID- 10482322 TI - Use of a control chart to monitor diarrhoea admissions: a quality improvement exercise in West Kalimantan Provincial Hospital, Pontianak, Indonesia. AB - Data on the number of admissions for diarrhoea each week to the West Kalimantan Provincial Hospital, Pontianak, Indonesia over a 5 year period, 1992-1996, were collected. After cleaning and exclusion of extreme values, transformation was then performed to ensure that the data were free of special cause variation and normally distributed. A control chart was then constructed to provide an 'early warning' system for hospital authorities in order to facilitate the management of the epidemic and to improve patient care. PMID- 10482323 TI - Outcome evaluation of early discharge of asthmatic children from hospital: a randomized control trial. AB - The objective of our study was to compare the safety and efficacy of discharging asthmatic children from hospital on three versus four hourly nebulized salbutamol. The setting was a tertiary referral paediatric hospital in Sydney, NSW, Australia. The design was a randomized controlled parallel group study. All children admitted to hospital with acute asthma and who were over 18 months of age were eligible to enter the study. Patients were excluded if they had non English speaking parents, no telephone, or chronic cardiac or neurological disease. Children were treated according to standard asthma management but were randomly allocated to be discharged on three or four hourly nebulized salbutamol. Patients were surveyed using a telephone questionnaire 1 to 2 weeks after discharge. The primary outcome measure was re-presentation to the Emergency Department (ED) within 7 days. Other outcomes included readmission to hospital, re-presentation to the local doctor, parental satisfaction and length of hospital stay. A total of 63 children were enrolled in the study (32 in the three hourly group and 31 in the four hourly group). There were no re-presentations to the ED or hospital readmissions within 1 to 2 weeks in either group. However, re presentations to the local doctor were common, 71.8% in the three hourly and 74.1% in the four hourly groups, respectively. These were predominantly for routine review. The mean (+/- SD) hospital length of stay was not significantly different between the three and four hourly groups, 48.94 (+/- 20.61) and 54.88 (+/- 32.59) hours, respectively (P = 0.672). Parents felt the timing of discharge was 'too early' in five (15.6%) of three hourly and five (16.1%) of four hourly patients. Three (9.7%) of the four hourly but none of the three hourly patients felt they were sent home 'later than necessary'. Five (15.1%) of the three hourly and three (9.7%) of the four hourly group parents did not feel comfortable looking after their child at home immediately after discharge. None of these differences were statistically significant. Discharge of asthmatic children from hospital on three hourly nebulized salbutamol is as safe and effective as on four hourly. Parents are generally very satisfied with timing of discharge, irrespective of frequency of nebulization. Earlier discharge benefits both the child and their family, and improves hospital bed utilization. PMID- 10482324 TI - Resuscitations in an observation unit. AB - The outcome of all resuscitations in a 20-bed observation unit of a large teaching hospital over a 25-month period was reviewed. Resuscitation was defined as a patient receiving one or more of the following: external chest compressions; defibrillation, assisted ventilation (e.g. intubation); and/or advanced cardiac life support drug therapy such as atropine or lidocaine for life threatening dysrhythmias including ventricular tachycardia, ventricular fibrillation, or asystole. There were nine patients out of 10,245 patients admitted to the observation unit (9/10,245 = 0.088% or approximately 0.09%) over the 26-month period from May 1994 to July 1996 who needed resuscitation. Each patient was initially successfully resuscitated. There was only one death (1/10,245 = 0.0098% or approximately 0.01%) in the observation unit. This patient was resuscitated with return of a spontaneous pulse and blood pressure only to expire while awaiting transfer to the Intensive Care Unit (ICU). Eight of the nine patients were admitted to the ICU, four of whom later died, making a total of five deaths (5/10,245 = 0.049% or approximately 0.05%). The other four patients were discharged neurologically intact and at 1 year follow-up were alive and doing well. Compared with resuscitation rates for the prehospital setting, the emergency department, the hospital medical/surgical floors, or the intensive care units, there is a higher successful resuscitation rate for the observation unit. PMID- 10482325 TI - The systematic assessment of short-term functional recovery after major joint arthroplasty. AB - Despite extensive information about long-term recovery from major joint arthroplasty, little attention has been given to the measurement of functional recovery in the immediate postoperative period. Therefore assumptions about the importance of physical therapy during this period remain untested. We devised a way of recording functional recovery before discharge, based primarily on the achievement of objective milestones. This was incorporated into routine physiotherapy practice and applied to sequential patients undergoing elective hip (n = 163) or knee (n = 66) replacement. Six months later, we followed up 160 patients, of whom 145 completed questionnaires to assess subjective physical and emotional state and functional recovery. The method was sensitive to known influences on pace of recovery, including type of arthroplasty (hip vs knee) and surgeons' differing requirements for mobilization. In addition, we were able to confirm and quantify sources of variation in functional recovery which previously were suspected but unconfirmed: in particular, the timing of early mobilization. Outcome at 6 months was unrelated to objective functional recovery in hospital, although fatigue and wellbeing at this time were predicted by physiotherapists' subjective assessment of patients' motivation before discharge. The findings can be used to inform patients and as a source of comparison data for the assessment of functional recovery in other centres. More importantly, the procedure reported may be applied to quantify functional recovery in routine practice and thereby expose variability in recovery to scientific scrutiny. PMID- 10482326 TI - Can a PC-based model assist the management of waiting lists? Observations from a case study. AB - Routine statistics provide only limited support to doctors and managers responsible for the management of surgical waiting lists. Therefore, a study developed a PC-based model that could assist medium-term planning activities that, for example, aim to reduce excessive waiting times. The model allows the user to create scenarios that describe possible future activity levels and calculates how these will affect waiting list behaviour. A key feature of the model is that it recognizes that a waiting list consists of different categories of patients that move through the list at different speeds. The model can be used with routinely available data. The model was tested at three sites where it proved capable of providing valuable insights and assisting doctors and managers to better understand waiting list dynamics. However, although the model quantified the changes required to reduce waiting times, the sites could not always implement these changes owing to resource constraints. PMID- 10482327 TI - Renal Home Support Scheme quality initiative. AB - In 1995 the practice of visiting home dialysis patients was challenged. In order to justify this practice, a benchmarking approach was adopted comprising a literature search, surveys of all relevant patients and their primary health carers, as well as all dialysis units in Australia. The results favoured the home visits and facilitated the establishment of a baseline standard of one home visit at least every 6 months. This standard was audited and reviewed annually. In 1996, Queensland Health launched the Renal Home Support Scheme which was targeted at improving the delivery of health care in the home, leading to efficiencies in the delivery of care to specific groups within the community such as the indigenous population. Successful submissions had to identify objectives, strategies and performance indicators that demonstrated the likelihood of success. The Renal Unit at Townsville General Hospital successfully tendered a submission to Queensland Health and the scheme was implemented in November 1996. PMID- 10482328 TI - The application of the continuum of care model in the re-configuration of nutrition and dietetics services. AB - The continuum of health care model can be used to improve service delivery. Within a hospital setting, finite resources are available. Service gaps must be identified, priorities established and resources re-distributed to meet these gaps. Using Continuous Quality Improvement (CQI) tools, barriers to change were identified and a plan for improvement was formulated. Areas receiving disproportionate resources were identified and a more even resource allocation adopted. In an illustration of the success of strategic change, core services were not only maintained but also showed improved efficiency. The Department of Nutrition and Dietetics at Wollongong and Port Kembla Hospitals achieved an increase in dietetic services through a reorientation of current resources.